PMID- 11409497 TI - Biodegradation of phenanthrene in soil. AB - We investigated the potential of an aerobic polycyclic aromatic hydrocarbon (PAH) adapted consortium to degrade phenanthrene in soil. Optimal degradation conditions were determined as pH7.0 and 30 degrees C with a water content of 100% wt soil/wt water (w/w). At a concentration of 5 microg/g, phenanthrene degradation (k1) was measured at 0.0269 l/hr with a half-life (t(1/2)) of 25.8 hrs. Our results show that the higher the phenanthrene concentration, the slower the degradation rates. Phenanthrene degradation was enhanced by treatment with yeast extract, glucose, or pyruvate, but was not significantly improved by the addition of acetate. Degradation was delayed by the addition of either compost or potassium nitrate and enhanced by the addition of nonionic surfactants (Brij30, Brij35, Triton X100 or Triton N101) at critical micelle concentration (CMC). Phenanthrene degradation was delayed at levels above CMC. PMID- 11409498 TI - Swine wastewater treatment using attached-growth and suspended-growth two stage sequencing batch reactors with real-time control. AB - Two two-stage sequencing batch reactors (TSSBR), one attached-growth and one suspended-growth, were operated under three levels of wastewater concentration (approximately 4,000, 2,000 and 500 TOC mg/L), respectively, to compare the pH and ORP (oxidation-reduction potential) patterns and system performance. In both TSSBR systems, the pH and ORP profiles varied with organic loading yet exhibited consistent patterns with distinctive features suitable for real-time control. For all runs at the three levels of influent, both systems achieved similar levels of treatment for BOD5, TOC and TSS of over 97.5, 93.4, and 97.3%, respectively. The attached-growth system out performed the suspended-growth system in achieving the same levels of treatment at much shorter aeration cycle times. The treatment efficiency for NO3(-)-N and PO4(-3) was greatly affected by the carbon content in the wastewater, and the best treatment was achieved during the TOC approximately 4,000 mg/L runs with final effluent at 4.0 and 21.3 mg/L, respectively. PMID- 11409499 TI - The effect of limited aeration on swine manure phosphorus removal. AB - Two low level aeration schemes (intermittent vs. continuous) were investigated on a laboratory scale, in conjunction with swine manure pH adjustment using sodium hydroxide (1.0 M), for manure phosphorus (P) removal. According to the data, an 80% reduction in soluble P was observed when the manure pH was increased to 8. Both intermittent and continuous aeration treatments could raise manure pH above 8 with an airflow rate of 1 L/minute in a period of 15 days. A drastic increase in pH (about 1 unit) was observed for both aeration schemes within the first day of test, resulting in a 76% reduction in soluble P concentration in the liquid. It appeared that there is no difference in terms of P removal between the two aeration programs, suggesting that the intermittent aeration be preferred to save energy while still achieving the same level of P removal. PMID- 11409500 TI - Response of foodborne Salmonella spp. marker strains inoculated on egg shell surfaces to disinfectants in a commercial egg washer. AB - The objective of this study was to determine the effectiveness of an iodine based disinfectant (IBD, Iocide, Biomedical Development Corporation, San Antonio, TX) on Salmonella enteritidis and S. typhimurium inoculated on egg shell surfaces under simulated industry egg processing conditions with a commercial egg washer used as the sanitizer delivery system. Re-circulated egg washer water containing 1.40-2.85 g/l total dissolved solids was obtained from a commercial egg processing. Sanitizing treatments consisted of distilled deionized water (DDW), IBD, and chlorine (CL; 200 ppm). All treatments (DDW, IBD and CL) significantly (p < 0.05) decreased Salmonella spp. populations on the shell compared to dry (no spray) egg controls. However, efficacy of egg sanitizers appeared to be dependent on the level of total dissolved solids in the egg wash water. PMID- 11409501 TI - Accumulation and fractionation of copper, iron, manganese, and zinc in calcareous soils amended with composts. AB - Amending soils with compost may lead to accumulation of metals and their fractions at various concentrations in the soil profile. The objectives of this study were to determine 1) the accumulation of Cu, Fe, Mn, and Zn with depth and 2) the distribution of water soluble, exchangeable, carbonate, Fe-Mn oxides, organic and residual forms of each metal in soils amended with MSW compost, co compost, biosolids compost and inorganic fertilizer (as control). Total concentrations of Cu, Fe, Mn and Zn were concentrated in the 0-22 cm soil layer and scant in the rock layer. These metals were in the decreasing order of Fe >> Mn > Zn > or = Cu. Copper, Fe, and Zn were predominantly in the residual form followed by fractions associated with Fe-Mn oxides, carbonate, organic, exchangeable and water soluble in all treatments except MSW compost amended soil where the organic fraction was higher than the carbonate fraction. In fertilizer, co-compost and biosolids compost treated soils Mn concentrated mainly in the Fe Mn oxides form followed by residual, carbonate, and organic forms whereas, in MSW compost treated soil the same pattern occurred except that Mn organic fraction was higher than that in the carbonate form. The MSW compost has a greater potential to be used as a soil amendment to supply plants with Cu, Mn and Zn than other treatments in calcareous soils of south Florida. PMID- 11409502 TI - Computerized brain-surface voltage topographic mapping for localization of intracranial spikes from electrocorticography. Technical note. AB - The purpose of this paper is to describe the use of computerized brain-surface voltage topographic mapping to localize and identify epileptic discharges recorded on electrocorticographic (ECoG) studies in which a subdural grid was used during intracranial video electroencephalographic (IVEEG) monitoring. The authors studied 12 children who underwent surgery for intractable extrahippocampal epilepsy. Cortical surfaces and subdural grid electrodes were photographed during the initial surgery to create an electrode map that could be superimposed onto a picture of the brain surface. Spikes were selected from ictal discharges recorded at the beginning of clinically confirmed seizures and from interictal discharges seen on ECoG studies during IVEEG recording. A computer program was used to calculate the sequential amplitude of the spikes by using squared interpolation, and they were then superimposed onto the electrode map. Interictal discharges and high-amplitude spike complexes at seizure onset were plotted on the map. This mapping procedure depicted the ictal zone in nine patients and the interictal zone in 12, and proved to be an accurate and useful source of information for planning corrective surgery. PMID- 11409503 TI - Revision of deep brain stimulator for tremor. Technical note. AB - The treatment of essential tremor with thalamic deep brain stimulation (DBS) is considered to be more effective and to cause less morbidity than treatment with thalamotomy. Nonetheless, implantation of an indwelling electrode, connectors, and a generator is associated with specific types of morbidity. The authors describe three patients who required revision of their DBS systems due to lead breakage. The connector between the DBS electrode and the extension wire, which connects to the subclavicular pulse generator, was originally placed subcutaneously in the cervical region to decrease the risk of erosion through the scalp and to improve cosmesis. Three patients presented with fractured DBS electrodes that were located in the cervical region near the connector, necessitating reoperation with stereotactic retargeting and placement of a new intracranial electrode. At reoperation, the connectors were placed subgaleally over the parietal region. Management of these cases has led to modifications in the operative procedure designed to improve the durability of DBS systems. The authors recommend that surgeons avoid placing the connection between the DBS electrode and the extension wire in the cervical region because patient movement can cause microfractures in the electrode. Such microfractures require intracranial revision, which may be associated with a higher risk of morbidity than the initial operation. The authors also recommend considering prophylactic relocation of the connectors from the cervical area to the subgaleal parietal region to decrease the risk of future DBS electrode fracture, which would necessitate a more lengthy procedure to revise the intracranial electrode. PMID- 11409504 TI - Meningeal neurosarcoidosis mimicking convexity meningioma. Case illustration. PMID- 11409505 TI - Brachial plexus palsy from nodular fasciitis with spontaneous recovery: implications for surgical management. Case illustration. PMID- 11409506 TI - Radiosurgery and trigeminal neuralgia. PMID- 11409507 TI - Shunt infection. PMID- 11409508 TI - Brain abscess. PMID- 11409509 TI - Germinoma and radiation. PMID- 11409510 TI - Subdural hematoma recurrence. PMID- 11409511 TI - Subdural hematoma recurrence. PMID- 11409512 TI - Potassium and intracranial pressure. PMID- 11409513 TI - Treatment of hydrocephalus in adults by placement of an open ventricular shunt. AB - OBJECT: Ventricular shunt placement is the neurosurgical procedure most frequently associated with complications. Over the years, it has been a growing concern that the performance of most shunting devices does not conform to physiological parameters. An open ventriculoperitoneal (VP) bypass with a peritoneal catheter for which the cross-sectional internal diameter was 0.51 mm as a distinctive element for flow resistance was evaluated for use in the treatment of adult patients with hydrocephalus. METHODS: During a 2-year period, open shunts were surgically implanted in 54 adults with hydrocephalus; conventional shunts were implanted in 80 matched controls. Periodic evaluations were performed using neuroimaging studies and measures of clinical status. All patients were followed from 12 to 36 months. 18.5 +/- 4 months for patients with the open shunt and 19.1 +/- 8.1 months for controls (mean +/- standard deviation). The device continued to function in 50 patients with the open shunt (93%) and in 49 controls (61%: p < 0.001). The Evans index in patients with the open shunt was 0.33 +/- 0.09 throughout the follow up. No cases of infection, overdrainage, or slit ventricles were observed: the index in controls was 0.28 +/ 0.08; 60% of them developed slit ventricles. During the follow-up period occlusion occurred in four patients with the open shunt (7%) and in 31 controls (39%: p < 0.001). CONCLUSIONS: The daily cerebrospinal fluid (CSF) drainage through the open VP shunt is close to 500 ml of uninterrupted flow propelled by the hydrokinetic force generated by the combination of ventricular pressure and siphoning effect. It complies with hydrokinetic parameters imposed by a bypass connection between the ventricular and peritoneal cavities as well as with the physiological archetype of continuous flow and drainage according to CSF production. The open shunt is simple, inexpensive, and an effective treatment for hydrocephalus in adults. PMID- 11409514 TI - Endovascular treatment of intracranial aneurysms by using Guglielmi detachable coils in awake patients: safety and feasibility. AB - OBJECT: Embolization of intracranial aneurysms performed using Guglielmi detachable coils (GDCs) is performed with the patient in a state of general anesthesia at most centers. Such an approach does not allow intraprocedural evaluation of the patient's neurological status and carries additional risks associated with general anesthesia and mechanical ventilation. At the authors' institution, GDC embolization of intracranial aneurysms is performed in awake patients after administration of sedative and analgesic agents (midazolam, fentanyl, morphine, and/or hydromorphone). To determine the feasibility and safety of this approach, the authors have retrospectively reviewed their clinical experience. METHODS: The authors reviewed the medical records of all patients in whom GDC embolization for the treatment of intracranial aneurysms was undertaken between February 1, 1990 and October 31, 1999. Clinical presentation, medical comorbidities, anesthetic agents used, intraprocedural complications, and final procedural outcome were recorded for each patient. Guglielmi detachable coil embolization was attempted in the awake patient in 150 procedures. Among 92 procedures for unruptured aneurysms, 75 (82%) were completed without complications. Four procedures were completed with complications. Of the 92 procedures, 13 were aborted due to patient uncooperativeness (one patient), complications (three patients), morphological characteristics of the aneurysm or surrounding vessels that made embolization technically difficult (eight patients), or vasospasm (one patient). Among 58 procedures for ruptured aneurysms, the procedure was completed without complication in 48 cases (83%). The procedure was completed with complications in five cases and two patients required induction of general anesthesia during the procedure. Five procedures were aborted because morphological characteristics of the aneurysm or surrounding vessels made embolization technically difficult (two patients) or because of aneurysm rupture (two patients) or the appearance of a transient neurological deficit (one patient). CONCLUSIONS: Embolization of intracranial aneurysms performed using GDCs in the awake patient appears to be safe and feasible and allows intraprocedural evaluation of the patient. Potential advantages, including decreased cardiopulmonary morbidity rates, shorter hospital stay, and lower hospital costs, still require confirmation by a direct comparison with other anesthetic procedures. PMID- 11409515 TI - Results of combined stereotactic radiosurgery and transarterial embolization for dural arteriovenous fistulas of the transverse and sigmoid sinuses. AB - OBJECT: Most dural arteriovenous fistulas (DAVFs) of the transverse and sigmoid sinuses do not have angiographically demonstrated features associated with intracranial hemorrhage and, therefore, may be treated nonsurgically. The authors report their experience using a staged combination of radiosurgery and transarterial embolization for treating DAVFs involving the transverse and sigmoid sinuses. METHODS: Between 1991 and 1998, 25 patients with DAVFs of the transverse and/or sigmoid sinuses were treated using stereotactic radiosurgery; 22 of these patients also underwent transarterial embolization. Two patients were lost to follow-up review. Clinical data, angiographic findings, and follow-up records for the remaining 23 patients were collected prospectively. The mean duration of clinical follow up after radiosurgery was 50 months (range 20-99 months). The 18 women and five men included in this series had a mean age of 57 years (range 33-79 years). Twenty-two (96%) of 23 patients presented with pulsatile tinnitus as the primary symptom; two patients had experienced an earlier intracerebral hemorrhage (ICH). Cognard classifications of the DAVFs included the following: I in 12 patients (52%), IIa in seven patients (30%), and III in four patients (17%). After treatment, symptoms resolved (20 patients) or improved significantly (two patients) in 96% of patients. One patient was clinically unchanged. No patient sustained an ICH or irradiation-related complication during the follow-up period. Seventeen patients underwent follow-up angiographic studies at a mean of 21 months after radiosurgery (range 11-38 months). Total or near-total obliteration (> 90%) was seen in 11 patients (65%), and more than a 50% reduction in six patients (35%). Two patients experienced recurrent tinnitus and underwent repeated radiosurgery and embolization at 21 and 38 months, respectively, after the first procedure. CONCLUSIONS: A staged combination of radiosurgery and transarterial embolization provides excellent symptom relief and a good angiographically verified cure rate for patients harboring low-risk DAVFs of the transverse and sigmoid sinuses. This combined approach is a safe and effective treatment strategy for patients without angiographically determined risk factors for hemorrhage and for elderly patients with significant comorbidities. PMID- 11409516 TI - Clinical and economic consequences of early discharge of patients following supratentorial stereotactic brain biopsy. AB - OBJECT: The goal of this study was to determine the clinical and economic consequences of early discharge (< 8 hours) of patients following stereotactic brain biopsy (SBB). METHODS: The records of all patients who underwent percutaneous SBB at The Cleveland Clinic Foundation, a tertiary care teaching hospital, during 1994 and 1995 (Group A) were retrospectively reviewed to collect data on the nature and timing of perioperative (< 48 hours) clinical and radiological complications. Biopsies were performed using image-guided stereotaxy either with or without a frame. Based on the results, guidelines for early discharge of patients following SBB were implemented. Information on the nature and timing of perioperative complications was also collected prospectively in all patients who underwent percutaneous SBB from January 1996 through July 1998 (Group B). Hospital financial records for patients who underwent SBB in 1997 and 1998 were also reviewed and assessed for net revenue stratified by discharge status: early discharge (< 8 hours), extended outpatient observation (> or = 8 and < 24 hours). and inpatient hospitalization (> or = 24 hours). In Group A, 130 biopsies were performed. There were five serious complications (3.8%), of which four were transient, and there was one death (0.8%). The death and any sustained deficit occurred in patients in whom a clot had been demonstrated on postoperative CT scans. All complications were detected within 6 hours after surgery. Intraoperative bleeding occurred in 12 patients (9.2%), but was associated with only 40% of cases in which hemorrhage appeared on postoperative CT scans. Guidelines for early discharge (< 8 hours) following SBB were developed and stipulated the absence of the following: 1) intraoperative hemorrhage; 2) new postoperative deficit; and 3) clot on a postoperative CT scan. In Group B, 139 biopsies were performed. There were three serious complications (2.2%), one of which was sustained due to a clot that had been demonstrated on the postoperative CT scan. All complications were detected within 6 hours postsurgery. There were no deaths in this group. Intraoperative bleeding occurred in 11 patients (7.9%), requiring intraoperative craniotomy to control bleeding in one case. Hospital financial records were available for 96 patients, of whom 22 were discharged from the hospital early, 11 were observed for an extended outpatient period, and the remainder were retained for inpatient hospitalization. Average net hospital incomes on technical charges for patients in the inpatient hospitalization, extended outpatient observation, and short-stay (early discharge) groups were $1778, $1175, and $1219, respectively, in 1997, but declined to -$889, -$1339, and $671, respectively, in 1998. The ratios of indirect costs to direct technical costs were 132.5%, 128.7%, and 103.7%, respectively. CONCLUSIONS: Early discharge of patients following SBB of supratentorial lesions is safe in the absence of excessive intraoperative bleeding, new postoperative deficit, and clot on a postoperative CT scan. Extended outpatient observation (8-23 hours) is not clinically necessary and may be economically prohibitive in the setting of a teaching hospital. PMID- 11409517 TI - Radiation necrosis following gamma knife surgery: a case-controlled comparison of treatment parameters and long-term clinical follow up. AB - OBJECT: Radiation necrosis is the only significant complication of gamma knife surgery (GKS). The authors studied treatment plan parameters in patients who had radiation necrosis to determine if risk factors for necrosis could be identified. METHODS: Between September 1994 and December 1998, 286 patients were treated with GKS by the senior author. Of the 243 patients who were suitable for analysis, 17 developed radiation necrosis and were prospectively followed. Concurrently, 17 patients without necrosis were randomly selected as case controls on the basis of histological findings in their lesions. Integral dose-volume histograms (DVHs) were calculated and dose-volume treatment parameters were determined. A comparison was made with both the established Kjellberg and Flickinger isonecrosis risk lines. Clinical outcome was assessed according to time to resolution of symptoms and return to normal radiographic appearance. CONCLUSIONS: Treatment plan variables associated with the risk of necrosis were increased tumor volume (TV) integral dose, increased TV, and increased 10-Gy volume. Other risk factors included repeated radiosurgery to the same lesion and glioma histological findings. The Kjellberg 1% risk line predicted a 5% risk of radiation necrosis and the Flickinger 3% risk line predicted a 3% risk. The median time to development of necrosis was 4 months, and symptomatic and radiographic recovery times were 7.5 and 10.5 months, respectively. The median survival time in patients with necrosis was 30 months. The authors recommend prospective TV determination and DVH calculation for all radiosurgical treatments and the avoidance of repeated radiosurgical treatments to the same lesion when possible. PMID- 11409518 TI - Evaluation of iatrogenic lesions in 722 surgically treated cases of peripheral nerve trauma. AB - OBJECT: The purpose of this study was to discover the number and types of iatrogenic nerve injuries that were surgically treated during a 9-year period at a relatively busy nerve center. The specific nerves involved, their sites of injury, and the mechanisms of injury were also documented. METHODS: The authors retrospectively evaluated the surgically treated iatrogenic lesions by reviewing case histories, operative reports, and follow-up notes in 722 cases of trauma. These cases were treated between January 1990 and December 1998 because of pain, dysesthesias, and sensory and/or motor deficits. latrogenic injury was a much larger category of trauma than predicted. One hundred twenty-six (17.4%) of the 722 surgically treated cases were iatrogenic in origin. Most of these injuries occurred during a previous operation. To a major extent, nerves of the extremities were affected, and a relatively large number of injuries occurred in the neck and groin. Incidence was highest in the spinal accessory nerve (14 cases), the common peroneal nerve (11 cases), the superficial radial nerve (10 cases), the genitofemoral nerve branches (10 cases), and the median nerve (nine cases). At least two thirds of the patients did not undergo surgery for the iatrogenic injury within an optimal time interval due to delayed referral. Follow up data were available in 97 of the 126 patients. Surgical outcomes demonstrated improvement in 70% of patients. Operative results were especially favorable in patients suffering from iatrogenic injuries to the accessory and superficial sensory radial nerves. CONCLUSIONS: latrogenic injuries should be corrected in a timely fashion just like any other traumatic injury to nerve. PMID- 11409519 TI - A 20-year review of percutaneous balloon compression of the trigeminal ganglion. AB - OBJECT: The aim of this study was to investigate outcomes and complication rates associated with percutaneous balloon compression (PBC) of the trigeminal ganglion over a long follow-up period. METHODS: This retrospective review was conducted in 496 patients with typical symptoms of unilateral trigeminal neuralgia who underwent 531 PBCs of the trigeminal ganglion between 1980 and 1999. The mean length of follow up was 10.7 years. The treatment used was a modification of that first described by Mullan and Lichtor in 1983. There were nine technical failures. Of the 522 successful procedures, prompt pain relief ensued in all patients except one. Recurrence of pain was found in 95 patients (19.2%) within 5 years and in 158 patients (31.9%) over the entire study period. Symptomatic dysesthesias occurred in 19 patients (3.8%), but corneal anesthesia and anesthesia dolorosa did not. CONCLUSIONS: In this review the authors present data on the largest cohort of patients with the longest follow up for this procedure in the current literature. The balance between the recurrence rate and troublesome sensory complications achieved in this series is favorable when compared with previously published studies on outcomes of PBC and the two alternative percutaneous methods, radiofrequency thermocoagulation and glycerol rhizolysis. The PBC procedure has additional advantages in that it is relatively straightforward and quick, and can be performed during a brief period of general anesthesia with no discomfort to the patient. This makes it an attractive first choice in the treatment of trigeminal neuralgia. PMID- 11409520 TI - Safety of perioperative minidose heparin in patients undergoing brain tumor surgery: a prospective, randomized, double-blind study. AB - OBJECT: Thromboembolic phenomena (TEPs) continue to be a significant source of morbidity and mortality in patients undergoing neurosurgery. Although the efficacy of low-dose heparin in preventing TEPs in neurosurgical patients is well established, neurosurgeons are reluctant to use it perioperatively because of concern for increased bleeding complications. To clarify this issue, the authors used a prospective, randomized, double-blind design to evaluate the safety of minidose heparin treatment in patients undergoing surgery for supratentorial brain tumors. METHODS: One hundred three patients, all 40 years of age or older, were treated with either 5000 U of heparin (55 patients) or placebo (48 patients) starting 2 hours before surgery and continuing until full mobilization or for 7 days. Both groups were well matched for sex, weight, duration of surgery, and tumor diagnosis. Subjective and objective parameters were used to estimate and calculate the perioperative bleeding tendency in all patients. Red blood cell mass loss was calculated by assessing the preoperative and postoperative hematocrit and the patient's weight. Intraoperative blood loss was determined by measuring the quantity of blood in the suction containers and subtracting the amount of irrigation fluids. Postoperative bleeding was measured by determining the amount of fluid in the subgaleal drain, and blood cell replacement was monitored during and after the procedure. Intracranial bleeding was graded according to findings on the postoperative computerized tomography scan obtained 48 to 72 hours after surgery. In addition, the senior surgeon in each case was asked to assess each patient's bleeding tendency during the operation. The results showed that perioperative administration of heparin did not significantly alter bleeding tendency by any measured parameter. The surgeon was blinded to which group individual patients had been allocated. CONCLUSIONS: Perioperative minidose heparin is safe for use in patients undergoing craniotomy for supratentorial tumors. This relatively simple and inexpensive measure is recommended as a routine regimen for the prevention of TEPs in patients undergoing neurosurgery. PMID- 11409521 TI - Determinants of tumor size and growth in vestibular schwannomas. AB - OBJECT: This study was undertaken to analyze factors associated with the size of unilateral vestibular schwannomas (VSs). METHODS: A retrospective analysis of an unselected and sequential series of 433 patients with unilateral VSs was conducted. Tumor size was defined by the largest dimension of the tumor in the cerebellopontine angle, and the size was tested for a relationship with patient age and sex. In a subgroup of 231 patients in whom data were available, tumor size was also tested for a relationship with tumor cysts or the absence of an internal auditory canal (IAC) component. Some patients underwent a period of surveillance with serial imaging studies to monitor for tumor growth. Data from these patients, excluding those with cystic tumors, were analyzed to see if tumor growth was related to patient age, sex, or tumor size. CONCLUSIONS; Larger tumors were found in younger patients, in females, in the subgroup of cystic tumors, and in patients in whom there was no tumor component in the IAC. The probable explanations for these larger tumors are a faster growth rate and/or a delay in symptom onset. When untreated tumors are managed with observation, measurable growth is more often seen in larger tumors, although smaller tumors have a faster relative growth rate than larger ones. PMID- 11409522 TI - Surgical treatment of posterior fossa tumors via the occipital transtentorial approach: evaluation of operative safety and results in 14 patients with anterosuperior cerebellar tumors. AB - OBJECT: To determine the safety and usefulness of performing surgery via the occipital transtentorial approach to treat anterosuperior cerebellar tumors, evaluation of 14 patients was performed over a 5-year period. METHODS: The study was performed in 14 patients, aged 6 months to 71 years, who harbored anterosuperior cerebellar tumors of the posterior fossa including four hemangioblastomas, three cerebellar astrocytomas, three medulloblastomas, two metastatic tumors, one recurrent astrocytoma, and one rhabdoid cell tumor. All patients underwent surgical treatment by the same surgical team and via the same surgical approach. Endoscopy combined with neuronavigation was used for large, deep-seated tumors extending to the fourth ventricle. Of the 14 patients, total or gross-total removal was achieved in 12 patients and subtotal removal in two patients. There was no incidence of mortality or morbidity in the 14 patients, and all functional outcomes were good to excellent postoperatively. Postoperative magnetic resonance imaging revealed that none of the patients had suffered brain damage or infarction around the cerebellum, brainstem, or occipital lobe. CONCLUSIONS: Although this study was the first in which a specific examination of the efficacy of the occipital transtentorial approach in patients with anterosuperior cerebellar tumors was undertaken, our findings suggest that this surgical approach is very useful, safe, and accurate for removing the primary tumor and evaluating the surrounding anatomy, as well as for determining operative strategy. PMID- 11409523 TI - Cranial surgery navigation aided by a compact intraoperative magnetic resonance imager. AB - OBJECT: In this article the authors report on a novel, compact device for magnetic resonance (MR) imaging that has been developed for use in a standard neurosurgical operating room. METHODS: The device includes a permanent magnet with a field strength of 0.12 tesla. The poles of the magnet are vertically aligned, with a gap of 25 cm. When not in use the magnet is stored in a shielded cage in a corner of the operating room; it is easily moved into position and attaches to a regular operating table. The magnet is raised for imaging when needed and may be lowered to allow surgery to proceed unencumbered. Surgical navigation with optical and/or magnetic probes is incorporated into the system. Twenty-five patients have undergone removal of intracranial lesions with the aid of this device. Operations included craniotomy for tumor or other lesion in 18 patients and transsphenoidal resection of tumor in seven. The number of scans ranged from two to five per surgery (average 3.4); image quality was excellent in 45%, adequate in 43%, and poor in 12%. In four patients MR imaging revealed additional tumor that was then resected; in five others visual examination of the operative field was inconclusive but complete tumor removal was confirmed on MR imaging. In 21 patients early postoperative diagnostic MR studies corroborated the findings on the final intraoperative MR image. Using a water-covered phantom, the accuracy of the navigational tools was assessed; 120 data points were measured. The accuracy of the magnetic probe averaged 1.3 mm and 2.1 mm in the coronal and axial planes, respectively; the optical probe accuracy was 2.1 mm and 1.8 mm in those planes. CONCLUSIONS: This device provides high-quality intraoperative imaging and accurate surgical navigation with minimal disruption in a standard neurosurgical operating room. PMID- 11409524 TI - Patterns of functional magnetic resonance imaging activation in association with structural lesions in the rolandic region: a classification system. AB - OBJECT: Functional magnetic resonance (fMR) imaging of the motor cortex is a potentially powerful tool in the preoperative planning of surgical procedures in and around the rolandic region. Little is known about the patterns of fMR imaging activation associated with various pathological lesions in that region or their relation to motor skills before surgical intervention. METHODS: Twenty-two control volunteers and 44 patients whose pathologies included arteriovenous malformations (AVMs; 16 patients), congenital cortical abnormalities (11 patients), and tumors (17 patients) were studied using fMR imaging and a hand motor task paradigm. Activation maps were constructed for each participant, and changes in position or amplitude of the motor activation on the lesion side were compared with the activation pattern obtained in the contralateral hemisphere. A classification scheme of plasticity (Grades 1-6) based on interhemispheric pixel asymmetry and displacement of activation was used to compare maps between patients, and relative to hand motor dexterity and/or weakness. There was 89.4% interobserver agreement on classification of patterns of fMR imaging activation. Displacement of activation by mass effect was more likely with tumors. Cortical malformations offer a much higher functional reorganization than AVMs or tumors. High-grade plasticity is recruited to compensate for severe motor impairment. CONCLUSIONS: Pattern modification of fMR imaging activation can be systematized in a classification of motor cortex plasticity. This classification has shown good correlation among grading, brain lesions, and motor skills. This proposal of a classification scheme, in addition to facilitating data collection and processing from different institutions, is well suited for comparing risks associated with surgical intervention and patterns of functional recovery in relation to preoperative fMR imaging categorization. Such studies are underway at the authors' institution. PMID- 11409525 TI - Three-dimensional anisotropy contrast magnetic resonance axonography to predict the prognosis for motor function in patients suffering from stroke. AB - OBJECT: The purpose of this study was to assess how early wallerian degeneration in the corticospinal tracts of patients who had suffered from stroke was detected using three-dimensional anisotropy contrast (3D-AC) magnetic resonance (MR) axonography and to explore the possibility of predicting the prognosis for motor function in these patients. METHODS: Ten healthy volunteers and 16 stroke patients with hemiparesis were studied using MR images including 3D-AC MR axonography images obtained using a 1.5-tesla MR imaging system. The axonography was performed using an echoplanar imaging method. All patients underwent MR studies 2, 3, and 10 weeks after stroke onset. To detect wallerian degeneration, the diffusion anisotropy in the corticospinal tracts at the level of the upper pons was evaluated on axial images. These MR findings were compared with the patients' motor functions, which were classified according to the Brunnstrom criteria 12 weeks after the onset of stroke. In all patients with poor recovery (Brunnstrom Stages I-IV), wallerian degeneration, which was demonstrated as a reduction in diffusion anisotropy on axonography images, could be observed in the corticospinal tracts; this degeneration was not found in patients with good recovery (Stages V and VI). Axonography could be used to detect degeneration between 2 and 3 weeks after stroke onset. On conventional T2-weighted MR images, hyperintense areas indicating wallerian degeneration were not detected until 10 weeks after stroke onset. CONCLUSIONS: With the aid of 3D-AC MR axonography, wallerian degeneration can be detected in the corticospinal tracts during the early stage of stroke (2-3 weeks after onset), much earlier than it can be detected using T2-weighted MR imaging. The procedure of 3D-AC MR axonography may be useful in predicting motor function prognosis in stroke patients. PMID- 11409526 TI - Evidence for telomerase involvement in the angiogenesis of astrocytic tumors: expression of human telomerase reverse transcriptase messenger RNA by vascular endothelial cells. AB - OBJECT: Evidence from recent in vitro studies indicates that reactivation of telomerase, the enzyme that synthesizes the telomere ends of chromosomes, is a crucial event in the unlimited clonal expansion of endothelial cells that precedes the neoplastic conversion of these cells. It is known that high-grade gliomas express telomerase and that, in these neoplasms, proliferating endothelial cells may undergo transformational changes with development of sarcomatous components within the primitive tumor. To assess whether telomerase is involved in the endothelial cell proliferation that characterizes brain tumor angiogenesis, the authors investigated at the single-cell level the expression of messenger (m)RNA for the human telomerase catalytic subunit human telomerase reverse transcriptase (hTERT) by vascular cells of astrocytic tumors. METHODS: The in situ hybridization (ISH) method was performed by processing histological sections with specific riboprobes for hTERT and for c-myc, an oncogene that is known to upregulate hTERT. Results of the ISH studies were compared with proliferative activity, as estimated by Ki-67 immunostaining. The expression of hTERT mRNA by vascular endothelial cells was related to the histological grade of the tumor because it was detected in five (29%) of 17 low-grade astrocytomas, nine (56%) of 16 anaplastic astrocytomas, and 19 (100%) of 19 glioblastomas multiforme (GBMs). Expression of c-myc mRNA was strictly correlated with that of hTERT mRNA. In low-grade astrocytomas and anaplastic astrocytomas, a dissociation was noted between hTERT mRNA expression and the proliferation rate of endothelial cells. Conversely, GBMs displayed a significant correlation between the level of hTERT mRNA expression and endothelial cell proliferation. Data from an in vitro assay in which human umbilical vein endothelial cells were stimulated to proliferate by adding vascular endothelial growth factor and an ISH study of newly formed vessels surrounding brain infarcts confirmed that expression of hTERT mRNA does not merely reflect the proliferative status of endothelial cells but represents a specific feature of brain tumor neovascularization. CONCLUSIONS: The results of this study are consistent with a role of telomerase in the angiogenesis of astrocytic tumors. Expression of hTERT mRNA by tumor vascular cells is an early event during the progression of astrocytic tumors, which precedes endothelial cell proliferation and may represent a first sign of dedifferentiation. Other than elucidating the mechanisms of tumor angiogenesis, these results encourage research on antitelomerase drugs for the treatment of malignant gliomas. PMID- 11409527 TI - Endothelial proliferation, neoangiogenesis, and potential de novo generation of cerebrovascular malformations. AB - OBJECT: To date, both arteriovenous malformations (AVMs) and cavernomas have been considered to be congenital malformations. A recent survey of the literature has shown the potential for de novo generation of both familial and sporadic cavernomas as well as AVMs. Therefore, it was of interest to determine the biological behavior of these lesions in detail. METHODS: The proliferative and angiogenic capacities of the endothelium of 13 cavernomas and 25 AVMs obtained in patients recently treated (1997-1998) at one institution were studied. Immunohistochemical staining for proliferating cell nuclear antigen (PCNA), MIB 1, and vascular endothelial growth factor (VEGF) and its receptor Flk-1 was performed using standard staining procedures. Positive immunostaining of the nuclei of endothelial cells was observed in specimens of both AVMs and cavernomas for PCNA (80% of AVMs and 85% of cavernomas), and Flk-1 (80% of AVMs and 31% of cavernomas). Endothelial expression of VEGF in the 18 incompletely embolized AVMs was found in 72% of cases but only in 28% of the seven cases in which patients did not undergo endovascular treatment: it was found in 38% of cavernomas. Endothelial expression of MIB-1 was found in 12% of AVMs but in no cavernomas. CONCLUSIONS: These results indicate that there is endothelial proliferation as well as neoangiogenesis in cerebral cavernomas and AVMs. The increased level of angiogenesis in only partially obliterated AVMs underscores the need for radical and complete occlusion of cerebral AVMs to avoid recurrences and further risks of morbidity. PMID- 11409528 TI - Molecular determinants of glioma cell migration and invasion. AB - OBJECT: Migration and invasion are important prerequisites for the infiltrative and destructive growth patterns of malignant gliomas. Infiltrative growth prevents complete tumor resection and causes significant neurological morbidity and mortality. METHODS: The authors assessed the expression of matrix metalloproteinases (MMPs) at messenger RNA and protein levels, MMP-2 and MMP-9 activities, and expression levels of a panel of anti- and proapoptotic proteins of the BCL-2 family. They then correlated their findings with alpha(v)beta3 integrin expression and the migratory and invasive potentials in 12 human malignant glioma cell lines. Multiple MMPs were expressed by most cell lines. The levels of MMP-2 and MMP-3 and the activities of MMP-2 and MMP-9 correlated with tumor cell invasion. Migration and invasion were also correlated. Although the expression levels of alpha(v)beta3 integrin did not predict migration or invasion, a neutralizing alpha(v)beta3 integrin antibody inhibited migration and invasion selectively in cell lines that contained a high level of alpha(v)beta3 integrin expression, thus indicating the important role of alpha(v)beta3 integrin for migration and invasion in this subset of cell lines. An expression pattern of BCL-2 family proteins that favor resistance to apoptosis was associated with enhanced migration, invasion, and MMP activity. Wild-type p53 cell lines migrated farther than mutant p53 cell lines. CONCLUSIONS: Activities of MMP-2 and MMP-9 are the best predictors of glioma cell invasion. The alpha(v)beta3 integrin mediates migration and invasion in a subset of glioma cell lines, but these processes do not depend on alpha(v)beta3 integrin expression. Antiapoptotic BCL-2 family protein expression is a predictor of efficient migration and invasion. PMID- 11409529 TI - Sagittal suture as an external landmark for the superior sagittal sinus. AB - OBJECT: Historically, the sagittal suture has been used as an external landmark to indicate the middle portion of the superior sagittal sinus (SSS). The goal of this study was to verify this relationship. METHODS: The authors examined 30 adult cadavers to reveal the location of the SSS with respect to the sagittal suture. Their findings demonstrated that the SSS is deviated to the right of the sagittal suture in the majority of observed specimens, although the maximum displacement to the right side was never more than 11 mm. CONCLUSIONS: This information should be useful to the neurosurgeon who must be aware of the SSS and its relationship with superficial skull landmarks. PMID- 11409530 TI - An unusual presentation of neurotuberculosis: subdural empyema. Case report. AB - Tuberculosis continues to be a major public health concern, especially in developing countries. Many types of neurotuberculosis have been described, but there is only one previously reported case of subdural empyema caused by tuberculous bacilli. A 1-year-old boy who had been treated for pulmonary tuberculosis was referred to the authors' institution with a diagnosis of right frontoparietal extraaxial abscess formation. Computerized tomography and magnetic resonance imaging revealed an extraaxial abscess with no evidence of calvarial infection. A craniotomy was performed to drain the pus, which was located subdurally. A polymerase chain reaction test yielded positive results, and histopathological examination revealed caseation. Antituberculous treatment was started after a diagnosis of subdural empyema with related neurotuberculosis had been made. At the end of a 12-month course of medical therapy, the patient was well with no evidence of tuberculosis. PMID- 11409531 TI - Intraoperative ultrasound arteriography with the "Coded Harmonic Angio" technique. Report of three cases. AB - The result of combining the ultrasound Coded Excitation method and an ultrasound contrast agent (UCA), the Coded Harmonic Angio (CHA) technique provides arterial images with exceptional spatial, temporal, and contrast resolution that are comparable to those produced by conventional digital subtraction angiography. The authors report on their experience with intraoperative ultrasound arteriography performed using the transdural CHA technique in three patients: one harboring a meningioma, another with a middle cerebral artery aneurysm, and a third with an arteriovenous malformation. The present study demonstrates how intraoperative cerebral ultrasound arteriography can be applied to assess the adequacy of neurovascular procedures without the presence of an experienced operator. PMID- 11409532 TI - External carotid artery reconstruction performed using an autologous internal carotid artery patch. Technical note. AB - The reconstruction of the external carotid artery (ECA) and obliteration of an occluded stump or atretic internal carotid artery (ICA) are described. In this method the occluded ICA is used as an autologous patch graft. Satisfactory ICA obliteration and expanded, smooth common carotid artery-ECA contour were obtained in three patients. Advantages of this procedure include ICA obliteration and precise patch placement by using an ideal, autologous, arterial graft. PMID- 11409533 TI - Extended transsphenoidal approach with submucosal posterior ethmoidectomy for parasellar tumors. Technical note. AB - The authors have developed an extended transsphenoidal approach with submucosal posterior ethmoidectomy for resection of tumors located in the cavernous sinus or the suprasellar region that are difficult to remove via the conventional transsphenoidal approach. Surgery was performed using this approach in 14 patients with large pituitary adenomas, three patients with craniopharyngiomas, and one patient with a meningioma of the tuberculum sellae. The submucosal dissection of the nasal septum used in the conventional transsphenoidal approach was extended to the superior lateral wall of the nasal cavity to expose the bony surface of the superior turbinate lying under the nasal mucosa. Submucosal posterior ethmoidectomy widened the area visualized through the conventional transsphenoidal approach both superiorly and laterally. This provided a safer and less invasive access to lesions in the cavernous sinus or the suprasellar region through the sphenoid sinus. Using this approach the authors encountered no postoperative complications, such as olfactory disturbance, cranial nerve palsy, or arterial injury. In this article the authors present the surgical methods used in this approach. PMID- 11409534 TI - Alternate cadmium exposure differentially affects the content of gamma aminobutyric acid (GABA) and taurine within the hypothalamus, median eminence, striatum and prefrontal cortex of male rats. AB - This work examines changes of gamma aminobutyric acid (GABA) and taurine contents in the hypothalamus, striatum and prefrontal cortex of the rat after an alternate schedule of cadmium administration. Age-associated changes were also evaluated, of those before puberty and after adult age. In control rats GABA content decreased with age in the median eminence and in anterior, mediobasal and posterior hypothalamus, prefrontal cortex and the striatum. Taurine content showed similar results with the exception of mediobasal hypothalamus and striatum, where no changes were detected. In pubertal rats treated with cadmium from 30 to 60 days of life, GABA content significantly decreased in all brain regions except in the striatum. When cadmium was administered from day 60 to 90 of life, GABA content was significantly changed in prefrontal cortex only compared with the age matched controls. Taurine content showed similar results in pubertal rats, with the exception of the median eminence and the mediobasal hypothalamus, neither of which showed a change. However, when cadmium was administered to rats from day 60 to 90 of life, taurine content only changed in prefrontal cortex compared with the age matched controls. These results suggest that cadmium differentially affects GABA and taurine contents within the hypothalamus, median eminence, striatum and prefrontal cortex as a function of age. PMID- 11409535 TI - Assessing interaction thresholds for trichloroethylene in combination with tetrachloroethylene and 1,1,1-trichloroethane using gas uptake studies and PBPK modeling. AB - The volatile organic solvents trichloroethylene (TCE), tetrachloroethylene (perchloroethylene, PERC), and 1,1,1-trichloroethane (methylchloroform, MC) are widely distributed environmental pollutants and common contaminants of many chemical waste sites. To investigate the mode of pharmacokinetic interactions among TCE, PERC, and MC and to calculate defined "interaction thresholds", gas uptake experiments were performed using a closed-chamber exposure system. In each experiment, two rats (Fischer 344, male, 8-9 weeks old) were exposed to different initial concentrations of TCE, PERC, and MC, applied singly or as a mixture, and their concentration in the gas phase of the chamber was monitored over a period of 6 h. A physiologically based pharmacokinetic (PBPK) model was developed to test multiple mechanisms of inhibitory interactions, i.e., competitive, non competitive, or uncompetitive. All mixture exposure data were accurately described by a system of equations in which a PBPK model was provided for each chemical and each was regarded as an inhibitor of the others' metabolism. Sensitivity-analysis techniques were used to investigate the impact of key parameters on model output and optimize experimental design. Model simulations indicated that, among these three chemicals, the inhibition was competitive. The PBPK model was extended to assess occupationally relevant exposures at or below the current threshold-limit values (TLVs). Based on 10% elevation in TCE blood levels as a criterion for significant interaction and assuming TCE exposure is set at TLV of 50 ppm, the calculated interaction thresholds for PERC and MC were 25 and 135 ppm, respectively. TLV exposures to binary TCE/PERC mixture were below the 10% significance level. The interaction threshold for TCE and MC co-exposure would be reached at 50 and 175 ppm, respectively. Such interactive PBPK models should be of value in risk assessment of occupational and environmental exposure to solvent mixtures. PMID- 11409536 TI - Biological monitoring of phenmedipham: determination of m-toluidine in urine. AB - Phenmedipham [methyl-3-(3-methylphenylcarbamoyloxy)carbamate] is used as a herbicide, especially in the growing of sugar beet and strawberries. During metabolism of the substance in rats, the two carbamate moieties of phenmedipham are cleaved and the metabolites methyl-N-(3-hydroxyphenyl)-carbamate, m aminophenol and hydroxyacetanilide are formed. These compounds and their conjugates are excreted in urine. Additionally, it has been suggested that m toluidine is formed during metabolism. For the first time it has been possible to detect this metabolite in the urine of workers after agricultural use of phenmedipham. The concentrations of m-toluidine in urine were significantly higher in persons occupationally exposed than in controls. The median values for each group were 0.36 microg/l and 0.16 microg/l, respectively. This means that persons not exposed to phenmedipham also excrete m-toluidine, possibly as a result of the uptake of pesticides like phenmedipham from the diet. PMID- 11409537 TI - Changes in antioxidant enzyme expression in response to hydrogen peroxide in rat astroglial cells. AB - Oxidative stress has been causally linked to a variety of neurodegenerative diseases. To clarify the role of the antioxidant enzyme (AOE) system in oxidative brain damage primary cultures of rat astroglial cells were exposed to hydrogen peroxide (H2O2). Expression of AOEs and several parameters for cell viability and functionality were measured. In our experiments astrocytes responded to low concentrations of H2O2 exposure with a pronounced generation of ROS which ran parallel with induction of lipid peroxidation. This distinct oxidative stress was not reflected in cell viability or functionality parameters measured. Cytotoxicity, a decrease in glutathione content of astrocytes, and impairment of mitochondrial functions became obvious only for higher concentrations of H2O2. After H2O2 exposure catalase, manganese superoxide dismutase, and glutathione peroxidase expression levels were found to be increased, whereas copper/zinc superoxide dismutase mRNA expression was not affected. These data indicate that the AOE system of astrocytes can be directly regulated by oxidative stress and may thus contribute to protection of cells against oxidative insults. PMID- 11409538 TI - Chronic pulmonary effects of respirable methylene diphenyl diisocyanate (MDI) aerosol in rats: combination of findings from two bioassays. AB - Two independent bioassays are available which have examined the potential carcinogenicity of monomeric and polymeric methylene diphenyl diisocyanate (MDI) following long-term inhalation exposure in rats. These studies are not directly comparable, however, due to differences in design and conduct of the in-life phase, and differences in nomenclature used for some of the histopathological findings. This paper presents a definitive overview ofthe pulmonary toxicity of MDI developed following a thorough review of both investigations. As part of this process, the test materials and the designs of the studies were compared, and an in-depth review of lung lesions was conducted by an independent reviewing pathologist. This included the re-examination of the original lung slides, supported by an analysis of the exposure regimens, the results of which were used to develop an accurate profile of the doses received by the animals in the two studies. Histopathological findings were then combined with this information to give an overall dose-response curve for both studies as a whole. The range of total inhalation exposures to MDI was calculated as 559, 1972, 2881, 6001, 17,575 and 17,728 mgh/m3. Major pulmonary effects included increased lung weights together with bronchiolo-alveolar adenomas and hyperplasia, and interstitial fibrosis which occurred consistently in both studies, indicating a very similar qualitative response of the lungs to polymeric and monomeric MDI. The quantitative response of the lung was clearly dose-related in each study, and when the studies were considered as a whole a reasonable overall dose-response relationship was apparent for major lung lesions. Lung tumours (in low incidences) only occurred at the highest dose level in both studies (17,575 and 17,728 mgh/m3). For inflammatory and other non-neoplastic pulmonary changes, the lowest dose examined (559 mgh/m3) was regarded as a no-observed-adverse-effect level for both polymeric and monomeric MDI. It was concluded that the results of the two studies could be combined to serve as a basis for human risk assessment of MDI. PMID- 11409539 TI - Aspartame prevents the karyomegaly induced by ochratoxin A in rat kidney. AB - Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus ochraceus as well as other moulds. This mycotoxin contaminates animal feed and food. OTA is immunosuppressive, genotoxic, teratogenic, carcinogenic and is nephrotoxic in all animal species studied so far. OTA inhibits protein synthesis and induces lipid peroxidation. Since it seems impossible to avoid completely contamination of foodstuffs by toxigenic fungi, it is necessary to investigate the possible ways of limiting such toxicity. An attempt to prevent OTA-induced nephrotoxic and genotoxic effects, mainly the karyomegaly, has been made in vivo using aspartame (L-aspartyl-L-phenylalanine methyl ester), a structural analogue of both OTA and phenylalanine. Aspartame (25 mg/kg body weight) prevented most of the nephrotoxic effects induced by OTA (289 microg/kg body weight). It also showed some utility in preventing morphological and histological damage, mainly the karyomegaly. The protective effects of aspartame on OTA-induced nephrotoxicity could be based on several mechanisms related to competitive binding to plasma proteins, to transport or tissue distribution in the kidney or to the elimination of the toxin in the urine. PMID- 11409540 TI - Tamoxifen-induced Ca2+ mobilization in bladder female transitional carcinoma cells. AB - This study examined the effect of tamoxifen, an anti-breast cancer drug, on Ca2+ handling in bladder female transitional cancer cells. Changes in cytosolic free Ca2+ levels were recorded by using the Ca2+-sensitive dye fura-2. In a dose dependent manner, tamoxifen induced intracellular free Ca2+ concentrations ([Ca2+]i) increases between 5 and 20 microM with an EC50 of 10 microM. External Ca2+ removal reduced the response by 60+/-6%. Addition of 3 mM Ca2+ caused a [Ca2+]i increase after pretreatment with 10 microM tamoxifen in Ca2+-free medium. In Ca2+-free medium, pretreatment with 10 microM tamoxifen abolished the [Ca2+]i increase induced by 1 microM thapsigargin, an endoplasmic reticulum Ca2+ pump inhibitor. Conversely, pretreatment with 1 microM thapsigargin prevented tamoxifen from releasing more Ca2+. Inhibition of phospholipase C-dependent inositol 1,4,5-tris-phosphate formation with 2 microM U73122 did not alter 10 microM tamoxifen-induced Ca2+ release. The [Ca2+]i increase induced by 5 microM tamoxifen was not altered by 10 microM La3+, nifedipine, verapamil, and diltiazem. Collectively, it was found that tamoxifen increased [Ca2+]i in bladder cancer cells by releasing Ca2+ from the endoplasmic reticulum Ca2+ stores in a manner independent of phospholipase C activity, and by inducing Ca2+ entry from external medium. PMID- 11409541 TI - A screen for sequences up-regulated during heterocyst development in Anabaena sp. strain PCC 7120. AB - A plasmid library of small genomic fragments from the cyanobacterium Anabaena sp. strain PCC 7120 was screened for sequences whose transcripts increase in abundance during a heterocyst development time course. A total of 350 clones were analyzed, representing 1-2% of the Anabaena sp. strain PCC 7120 genome. Twenty seven clones (8%) showed some degree of up-regulation after nitrogen starvation. The increase in transcript abundance ranged from 1.2-fold to 3.5-fold. Further analysis of the expression of some of the sequences using Northern blots suggested that the up-regulation values calculated from the screen are underestimates. The collection of up-regulated clones includes novel genes, previously characterized genes, and genes identifiable by similarity to known genes. One of the novel genes has been shown to be required for heterocyst function, and the sequence similarities and expression patterns of some of the others suggest that they may play a role in heterocyst development. PMID- 11409542 TI - ISZm1068: an IS5-like insertion element from Zymomonas mobilis. AB - A new insertion sequence, designated ISZm1068, was isolated from Zymomonas mobilis strain CP4. This element consists of 1,068 bp and contains one major ORF which shows similarities both at the nucleotide and at the amino acid sequence level with the corresponding ORFs encoding the transposases of many IS5 family elements, in particular the IS1031 group. Moreover, the Z. mobilis ORF shares the conserved N2, N3 and C1 signature motifs of the IS4 and IS5 families. Six out of seven Z. mobilis wild-type strains were shown by hybridisation to contain a single copy of the ISZm1068 element. Nucleotide sequences of the insertion elements from these strains exhibited extremely high levels of identity, varying from 94.25 to 99.25%. ISZm1068 was shown to be active in Escherichia coli cells and led to plasmid replicon fusions within the host cell. Sequence analysis of rare cointegration and resolution derivatives suggests that ISZm1068 has putative imperfect inverted repeats at its extremities of 18 bp (IR-right) and 14 bp (IR left), and that a 3-bp (5'-TCA-3') target sequence is duplicated upon insertion. PMID- 11409543 TI - Genetic and functional characterization of dpp genes encoding a dipeptide transport system in Lactococcus lactis. AB - The genes encoding a binding-protein-dependent ABC transporter for dipeptides (Dpp) were identified in Lactococcus lactis subsp. cremoris MG1363. Two (dppA and dppP) of the six ORFs (dppAdppPBCDF) encode proteins that are homologous to peptide- and pheromone-binding proteins. The dppP gene contains a chain terminating nonsense mutation and a frame-shift that may impair its function. The functionality of the dpp genes was proven by the construction of disruption mutants via homologous recombination. The expression of DppA and various other components of the proteolytic system was studied in synthetic and peptide-rich media and by using isogenic peptide-transport mutants that are defective in one or more systems (Opp, DtpT, and/or Dpp). In peptide-rich medium, DppA was maximally expressed in mutants lacking Opp and DtpT. DppA expression also depended on the growth phase and was repressed by tri-leucine and tri-valine. The effect of tri-leucine on DppA expression was abolished when leucine was present in the medium. Importantly, the Dpp system also regulated the expression of other components of the proteolytic system. This regulation was achieved via the internalization of di-valine, which caused a 30-50% inhibition in the expression of the proteinase PrtP and the peptidases PepN and PepC. Similar to the regulation of DppA, the repressing effect was no longer observed when high concentrations of valine were present. The intricate regulation of the components of the proteolytic system by peptides and amino acids is discussed in the light of the new and published data. PMID- 11409544 TI - Identification and characterization of a periplasmic nitrate reductase in Azospirillum brasilense Sp245. AB - The Azospirillum brasilense Sp245 napABC genes, encoding nitrate reductase activity, were isolated and sequenced. The derived protein sequences are very similar throughout the whole Nap segment to the NapABC protein sequences of Escherichia coli, Pseudomonas sp. G-179, Ralstonia eutropha, Rhodobacter sphaeroides, and Paracoccus denitrificans. Based on whole-cell nitrate reductase assays with the artificial electron donors benzyl viologen and methyl viologen, and assays with periplasmic cell-free extracts, it was concluded that the napABC encoded enzyme activity in Azospirillum brasilense Sp245 corresponds to a periplasmic dissimilatory nitrate reductase, which was expressed under anoxic conditions and oxic conditions. A kanamycin-resistant Azospirillum brasilense Sp245 napA insertion mutant was constructed. The mutant still expressed assimilatory nitrate reductase activity, but was devoid of its periplasmic dissimilatory nitrate reductase activity. PMID- 11409545 TI - Biotransformation of crotonobetaine to L(-)-carnitine in Proteus sp. AB - Two proteins, component I (CI) and component II (CII), catalyze the biotransformation of crotonobetaine to L(-)-carnitine in Proteus sp. CI was purified to electrophoretic homogeneity from cell-free extracts of Proteus sp. The N-terminal amino acid sequence of CI showed high similarity (80%) to the caiB gene product from Escherichia coli O44K74, which encodes the L(-)-carnitine dehydratase. CI alone was unable to convert crotonobetaine into L(-)-carnitine even in the presence of the cosubstrates crotonobetainyl-CoA or gamma butyrobetainyl-CoA, which are essential for this biotransformation. The relative molecular mass of CI was determined to be 91.1 kDa. CI is composed of two identical subunits of molecular mass 43.6 kDa. The isoelectric point is 5.0. CII was purified to electrophoretic homogeneity from cell-free extracts of Proteus sp. and its N-terminal amino acid sequence showed high similarity (75%) to the caiD gene product of E. coli O44K74. The relative molecular mass of CII was shown to be 88.0 kDa, and CII is composed of three identical subunits of molecular mass 30.1 kDa. The isoelectric point of CII is 4.9. For the biotransformation of crotonobetaine to L(-)-carnitine, the presence of CI, CII, and a cosubstrate (crotonobetainyl-CoA or gamma-butyrobetainyl-CoA) were shown to be essential. PMID- 11409546 TI - Mechanisms of acetate formation and acetate activation in halophilic archaea. AB - The halophilic archaea Halococcus (Hc.) saccharolyticus, Haloferax (Hf.) volcanii, and Halorubrum (Hr.) saccharovorum were found to generate acetate during growth on glucose and to utilize acetate as a growth substrate. The mechanisms of acetate formation from acetyl-CoA and of acetate activation to acetyl-CoA were studied. Hc. saccharolyticus, exponentially growing on complex medium with glucose, formed acetate and contained ADP-forming acetyl-CoA synthetase (ADP-ACS) rather than acetate kinase and phosphate acetyltransferase or AMP-forming acetyl-CoA synthetase. In the stationary phase, the excreted acetate was completely consumed, and cells contained AMP-forming acetyl-CoA synthetase (AMP-ACS) and a significantly reduced ADP-ACS activity. Hc. saccharolyticus, grown on acetate as carbon and energy source, contained only AMP ACS rather than ADP-ACS or acetate kinase. Cell suspensions of Hc. saccharolyticus metabolized acetate only when they contained AMP-ACS activity, i.e., when they were obtained after growth on acetate or from the stationary phase after growth on glucose. Suspensions of exponential glucose-grown cells, containing only ADP-ACS but not AMP-ACS, did not consume acetate. Similar results were obtained for the phylogenetic distantly related halophilic archaea Hf. volcanii and Hf. saccharovorum. We conclude that, in halophilic archaea, the formation of acetate from acetyl-CoA is catalyzed by ADP-ACS, whereas the activation of acetate to acetyl-CoA is mediated by an inducible AMP-ACS. PMID- 11409547 TI - Alkalispirillum mobile gen. nov., spec. nov., an alkaliphilic non-phototrophic member of the Ectothiorhodospiraceae. AB - From cultures of the anoxygenic phototroph Halorhodospira halophila SL-1, an aerobic, gram-negative spirillum was isolated. This moderately halophilic, alkaliphilic bacterium was motile by means of a single polar flagellum. It is described here as Alkalispirillum mobile gen. nov., spec. nov. Phylogenetic analysis of the Alkalispirillum mobile 16S rRNA gene led to its classification in the gamma-subclass of the Proteobacteria, as it appears closely related to phototrophic purple sulfur bacteria of the genera Ectothiorhodospira and Halorhodospira. Surprisingly, A. mobile is an obligate aerobe. The organism grows optimally with a number of carboxylic acids (such as sodium acetate) as carbon source, at 2% (i.e. approximately 0.34 M) sodium chloride, at pH 9-10, and at temperatures ranging from 35 to 38 degrees C. The dominant cellular fatty acids of Alkalispirillum mobile are C12:0, C16:0, C18:1cis11, and C18:0; its G+C content is 66.2+/-0.5 mol%. PMID- 11409548 TI - Isolation and characterization of a veratrol:corrinoid protein methyl transferase from Acetobacterium dehalogenans. AB - From 3-methoxyphenol-grown cells of Acetobacterium dehalogenans, an inducible enzyme was purified that mediated the transfer of the methyl groups of veratrol (1,2-dimethoxybenzene) to a corrinoid protein enriched from the same cells. In this reaction, veratrol was converted via 2-methoxyphenol to 1,2 dihydroxybenzene. The veratrol:corrinoid protein methyl transferase, designated MTIver, had an apparent molecular mass of about 32 kDa. With respect to the N terminal amino acid sequence and other characteristics, MTIver is different from the vanillate:corrinoid protein methyl transferase (MTIvan) isolated earlier from the same bacterium. For the methyl transfer from veratrol to tetrahydrofolate, two additional protein fractions were required, one of which contained a corrinoid protein. This protein was not identical with the corrinoid protein of the vanillate O-demethylase system. However, the latter corrinoid protein could also serve as methyl acceptor for the veratrol:corrinoid protein methyl transferase. MTIver catalyzed the demethylation of veratrol, 3,4 dimethoxybenzoate, 2-methoxyphenol, and 3-methoxyphenol. Vanillate (3-methoxy-4 hydroxybenzoate), 2-methoxybenzoate, or 4-methoxybenzoate could not serve as substrates. PMID- 11409549 TI - Study of pyoverdine type and production by Pseudomonas aeruginosa isolated from cystic fibrosis patients: prevalence of type II pyoverdine isolates and accumulation of pyoverdine-negative mutations. AB - The lungs of cystic fibrosis patients are frequently colonized by Pseudomonas aeruginosa, which produces high-affinity fluorescent peptidic siderophores, pyoverdines. Three pyoverdines which differ in their peptide chain and are easily differentiated by isoelectric focusing exist, only one being produced by a given strain. P. aeruginosa isolates from cystic fibrosis patients of a German hospital were analyzed by sequential, pulse-field gel electrophoresis (PFGE) and for pyoverdine production and type. Only producers of type I and type II pyoverdine were found. There was a perfect correlation between the type of pyoverdine produced and the clonality determined by PFGE. PFGE clone C, the most prevalent among cystic fibrosis patients, and found in an aquatic environment, produced type II pyoverdine. Pyoverdine-negative mutants seemed to increase as a function of the lung colonization time, but retained the capacity to take up pyoverdines. Most isolates that took up type II pyoverdine were also able to utilize type I pyoverdine as judged by growth stimulation experiments. No correlation was observed between the loss of pyoverdine production and mucoidy. PMID- 11409550 TI - Open versus arthroscopic treatment of chronic rotator cuff impingement. AB - We report the results of 238 consecutive patients who underwent in total 261 acromioplasties because of chronic rotator cuff impingement. The procedure was performed either in conventional open technique (80) or arthroscopically (181). Two years (1-10) after the operation 68% of the patients treated with the open technique had an excellent or good result compared 82% of the patients treated arthroscopically by an experienced arthroscopic surgeon. Compared to the open technique, the arthroscopic procedure had a statistically significant superior result concerning outcome, operating time and hospital stay. Arthroscopic procedures performed by less experienced surgeons had inferior results. PMID- 11409551 TI - Open wedge tibial osteotomy by hemicallotasis for medial compartment osteoarthritis. AB - Early clinical results were evaluated for 22 adult patients who had undergone an open wedge tibial osteotomy by hemicallotasis (HCO) due to medial compartment osteoarthritis. The mean age at the time of the index operation was 56 (range 33 66) years. The mean duration of follow up was 16 (range 9-23) months. The median hip-knee-ankle angle (HKA) was 169 (range 162-186) deg preoperatively and 182 (range 175-191) deg at follow up. We did not observe any early collapse of the new bone wedge. The median time to fixation was 79 (range 63-125) days. Complications included two pintract infections, and two hematomas were revised. Two patients felt pain during the phase of distraction, but the procedure could be continued after a short break. We evaluated the clinical results on the HSS, Lysholm, and Tegner activity scores. At the latest follow up examination, all of the scores had improved. Our findings demonstrate that HCO requires an exact correction, is a simple technique, and appears to reduce the chance of nerve and vascular damage. PMID- 11409552 TI - Autologous osteochondral transplantation using the diamond bone-cutting system (DBCS): 6-12 years' follow-up of 35 patients with osteochondral defects at the knee joint. AB - Thirty-five patients with severe osteochondral defects were treated by autologous osteochondral transplantation between 1986 and 1992. The majority of patients (27) suffered from osteochondrosis dissecans, while 8 patients presented with posttraumatic osteochondral defects. The grafts were harvested with a diamond bone cutter from the posterior part of the medial or lateral femoral condyle. In 29 patients the lesion was located at the lateral part of the medial femoral condyle, in 3 it was at the lateral femoral condyle, and in 3 at the patella. Twenty-nine patients could be examined at the follow-up between 6 and 12 years later (mean follow up 8.1 years). Using the standard cartilage evaluation form, the transplanted knees of 12 patients were graded as normal (grade I), 14 knees were nearly normal (grade II), while 3 patients presented with an abnormal result (grade III). All 3 of them had a varus malalignment and refused a high tibial correction osteotomy against our advice. No patient was assessed as severely abnormal (grade IV). The majority of patients improved their activity level and the functional status of the joint. Twelve patients developed new radiological signs of osteoarthrosis with a decrease in the radiological score of Kellgren and Lawrence by about one stage. We conclude that autologous osteochondral transplantation with the diamond bone-cutting system is an effective method in the treatment of severe osteochondral defects. PMID- 11409553 TI - The droopy shoulder syndrome. AB - Droopy shoulder syndrome (DSS) is characterized by a depression of the shoulders that stretches the brachial plexus, thus causing pain without any signs of neurological impairment. We describe ten patients with DSS; all had been treated for different diagnoses before. Contrary to previous reports, three patients had unilateral involvement, and five had accompanying disease of the cervical shoulder region. All patients responded well to conservative treatment in 2-10 weeks. DSS must be kept in mind in the differential diagnosis of pain in the cervical-shoulder region, to prevent unnecessary medication. PMID- 11409554 TI - Dementia does not significantly affect complications and functional gain in elderly patients operated on for intracapsular hip fracture. AB - Fifty-three elderly patients with intracapsular hip fracture were hospitalized in the Department of Orthopaedics. Forty-two underwent operations: 38 hemiarthroplasty; 2 total hip replacements and 2 closed reduction interior fixation. They were followed up and assessed prospectively five times for 6 months post-fracture. Cognition was evaluated by the Mini-Mental State Examination. Pre-fracture functioning was determined by the Functional Independence Measure (FIM) and the Katz index of activities of daily living (ADL). The functional outcome was assessed by the FIM gain defined as the difference between FIM scores at 6 months and just prior to discharge. FIM gain, length of stay, complications and mortality rates were not significantly different between three cognitive groups: normal, moderately and severely demented patients. The majority of patients were independent and partially dependent in their ADL. We conclude that dementia does not significantly affect complications and functional gain in elderly patients operated on for intracapsular hip fracture if they were mobile before the fracture. PMID- 11409555 TI - Causes of the clinical tissue response to polyglycolide and polylactide implants with an emphasis on the knee. AB - The causes of a clinical tissue response to self-reinforced polyglycolide (SR PGA) and self-reinforced polylactide (SR-PLLA) implants were studied in the first 2500 polymeric fixations. The incidence was 8.0% in fixations with implants made of SR-PGA (n = 1879) and 0% in SR-PLLA fixations (n = 621). A small number of fixations per site seemed to increase the incidence of a tissue response. In diagnoses involving over 50 fixations, except for ankle fractures, the incidence was 4.2%. The site of fixation influenced the incidence; the lowest incidence was seen for radial head fracture fixation (1.1%). The aromatic dye which has not been used since 1988 in the SR-PGA implant increased the response. A large total implant volume and a large number of implants increased the incidence of a tissue response. In the knee (n = 131), the incidence of a clinical tissue response was 4.3%, with four fluid accumulations and one synovitis of the knee after fixation with SR-PGA implants. PMID- 11409556 TI - An improved surgical technique to treat femoral shaft malunion: revised reamed intramedullary nailing technique. AB - Twenty-four consecutive adult patients who sustained a femoral shaft angular or rotational deformity, with or without significant shortening, were treated with skeletal traction of the femoral condyle, transverse femoral osteotomy, realignment of the shaft axis with or without lengthening, stable reamed intramedullary nail stabilization, and corticocancellous bone grafting. Twenty one patients who underwent the treatment were followed-up for at least 1 year (range 1.0-4.5 years), and all achieved a solid union. The union rate was 100% (21/21) with a union period of 4.9+/-0.5 months. There were no significant complications. All patients achieved a satisfactory outcome as indicated by patient satisfaction and physician assessment of outcome (p < 0.001, Fisher's exact test). In the present series, the use of this revised reamed nailing technique resulted in a high success rate and a low complication rate. It provides an effective alternative treatment in indicated cases. PMID- 11409557 TI - Preoperative radiotherapy without femoral shielding for prevention of heterotopic ossification in hydroxyapatite-coated hip prostheses. AB - Twenty-seven consecutive patients at high risk of developing heterotopic ossifications (HO) after implantation of a hydroxyapatite (HA)-coated hip prosthesis were irradiated with a single dose of 7 Gy, at least 4 h before the operation. The femoral stem was not shielded during radiotherapy (RT). After a median follow-up of 14.8 months, no clinically significant HO could be found, while 12 (52%) patients in this high-risk population had only minor HO (grade I). No reoperation was needed, and no evidence of prosthesis migration was observed. We conclude that single-dose, preoperative RT for HA-coated hip prosthesis can effectively inhibit HO. Not blocking the femoral stem does not result in prosthesis migration. PMID- 11409558 TI - Cement removal with an endoscopically controlled ballistically driven chiselling system. A new device for cement removal and preliminary clinical results. AB - In this paper, we introduce a newly developed miniaturised chiselling system (Swiss OrthoClast) which allows endoscopically controlled cement removal even from deep within the femur in hip revision arthroplasty. The intraoperative handling is demonstrated and illustrated. The device consists of a control unit, a handpiece, a set of chisels of different lengths, shape, and tip geometry, a set of plug extraction tools, and a special intrafemoral endoscope. It uses simple ballistic principles to effect fracturing of the bone cement. Inside the handpiece the projectile is accelerated and strikes the chisel head. The tip moves at high speed and with a small excursion towards the cement, which is fragmented mechanically by the movement of the chisel tip without generating heat. The device is being validated in a prospective, international, multicenter study, and we include here the results of the first 28 clinical procedures of cement removal. The duration of the procedure varied from 13 to 75 min, with an average of 32 min. In all but one case, the cement was completely removed. We noted one case of cortical fissuring. PMID- 11409559 TI - Postoperative analgesic value of the intra-articular instillation of bupivacaine and morphine after arthroscopic knee surgery. AB - This prospective, randomized, double-blind study was carried out with 320 patients, subdivided into 4 groups of 80 patients each, who underwent arthroscopic knee surgery during the time period from March 1995 to February 1997. Each of the 4 groups received a different solution: 15 ml of bupivacaine 0.5%, 5 mg of morphine in 15 ml of isotonic saline solution, 15 ml of bupivacaine 0.5% with epinephrine 0.0005%, or 15 ml of isotonic saline solution (control group). Within each study group, the following operations were performed: 15 cruciate ligament plasties with autologous patellar tendon grafts, 30 meniscus resections, 15 notch-plasties as preparation for anterior cruciate ligament surgery, and 20 other types of operation (plica resection, diagnostic biopsy, etc.). Pain intensity was assessed with the visual analogue scale obtained from questioning patients at 2, 4, 8, 12, and 24 h postoperatively. In terms of reducing postoperative pain and decreasing the consumption of analgesics after arthroscopic knee surgery, bupivacaine 0.5% with epinephrine 0.0005% was found to be the most effective. PMID- 11409560 TI - Distal femoral bone mineral density after total knee arthroplasty: a comparison with general bone mineral density. AB - The bone mineral density (BMD) of the distal femur may decrease after cemented total knee arthroplasty (TKA) as a result of the stress shielding effect of the femoral component. The purpose of the study was to determine the changes in BMD of the distal femur compared with those of the femoral necks and the lumbar spine after cemented TKA. BMD of two regions of interest in the distal femur, both femoral necks and the lumbar spine was measured with dual-energy X-ray absorptiometry in 10 patients (age range 41-80 years, mean 62 years) with 12 TKAs preoperatively and during follow-up for 1 year after surgery. The hip and spine measurements were performed for comparison to assess if general changes in BMD occurred after TKA. The median decrease in BMD in the region behind the anterior flange of the femoral component was 22% (95% CI: 12%-33%), while the average decrease in the region just above the femoral component was 8% (95% CI: 2%-13%). The difference in change of BMD between both regions before and 1 year after TKA was significant (p = 0.03). We found less than 1% difference in BMD of both femoral necks and the lumbar spine on average between the preoperative and 1 year follow-up measurements (not significant). A significant periprosthetic distal femoral bone resorption occurred after TKA. BMD of the femoral necks and lumbar spine did not differ 1 year after TKA. PMID- 11409561 TI - In-vitro reconstruction of massive rotator cuff ruptures with triceps tendon or coracoacromial ligament. AB - The possibility of reconstructing a massive rotator cuff rupture with a free graft of the triceps tendon or the coracoacromial ligament was tested experimentally in an anatomical and biophysical cadaver investigation. After dissection of the rotator cuff, a section of the supraspinatus tendon was removed to simulate a defect in the cuff. In dissection of the grafts, a bony squama was incorporated by osteotomy. The rotator cuff was reconstructed by suturing the free grafts into the defect. In addition, the bony squama was refixed in a prepared bone groove in the greater tuberosity of the humerus. The tensile strength and the elasticity of the grafts were compared with that of the supraspinatus tendon in rupture tests. The mean value of the maximum load determined (FRm) was greatest for the triceps tendon (706.451 N), while very much lower values were found for the coracoacromial ligament (395.836 N) and the supraspinatus tendon (496.792 N). The mean value of the maximum elasticity (LAm) was lowest for the coracoacromial ligament (4.701%); the supraspinatus tendon (7.047%) and the triceps tendon (8.250%) showed very much greater elasticity. PMID- 11409562 TI - Chronic persistent knee pain after repeated diagnostic arthroscopy: osteoid osteoma, a primarily overlooked diagnosis in a young woman. AB - We report a case of osteoid osteoma of the distal femur in an adolescent female patient that was primarily overlooked and led to repeated diagnostic arthroscopy of the knee. A thorough history and physical examination of the joint as well as adequate roentgenographic studies could have spared this patient two unnecessary arthroscopies and indicated the correct therapy. PMID- 11409563 TI - Malignant mixed tumor of salivary gland with a solitary metastasis to the tibia. AB - A 23-year-old, male patient previously operated on four times for a recurring, benign, paranasal mass was admitted to our clinic with pain and swelling under the knee. Physical and radiological examination revealed a malignant process. The histopathological examination of the fine-needle biopsy specimen of the tibia was a malignant mixed tumor. Then. en-bloc resection, free fibular grafting, and fixation with blade plate was performed for one-third of the tibia, conserving the knee joint. Recurrences and metastasis were not observed after surgery and radiotherapy. PMID- 11409564 TI - Treatment of staphylococcal implant infection with rifampicin-ciprofloxacin in stable implants. AB - Infection following total joint replacement remains a problem that has not been solved so far. The treatment options include removal of the implant and a delayed reconstruction or a direct exchange operation. Among patients with stable implants and short duration of infection as well as in patients who for certain reasons are inoperable, antibiotic therapy with a combination of rifampicin ciprofloxacin may be a reasonable treatment option for curing staphyloccocal infection without removal of the implant. A case study of a Staphylococcus epidermidis (coagulase-negative) infection following delayed revision total knee replacement after septic loosening of a knee arthroplasty and its successful conservative treatment with rifampicin-ciprofloxacin is described. Alternative rifampicin combinations are discussed with respect to recently developed pharmacodynamical and pharmacokinetical findings of biofilm active drugs. PMID- 11409565 TI - Relationship between DNA ploidy-related parameters and the deletions in mismatch repair genes MLH1 and MSH2 in lentigo maligna and malignant melanomas. AB - Defects in DNA mismatch repair genes MLH1 and MSH2, first described in hereditary nonpolyposis colon cancer (HNPCC), have been postulated to be responsible for malignant transformation in several tumours. To date there are no data on cutaneous tumours. Using a PCR assay it was possible to identify deletions in MSH2 (exonic regions 12 and 13) in 16 of 47 lentigos maligna and in 10 of 36 malignant melanomas. Deletions in MLH1 (exonic regions 15 and 16) were found in 11 of 47 lentigos and in 15 of 36 melanomas. Comparison of DNA ploidy-related parameters between lentigos with and without exonic deletions in MSH2 and MLH1 did not show any significant differences. In contrast, melanomas positive and negative for exons 12 and 13 (MSH2) (26/36 and 10/36, respectively) differed significantly with respect to the percentages of diploid cells (P = 0.0179) and tetraploid cells (P = 0.0042). Comparison of melanomas positive and negative for exons 15 and 16 (MLH1) (21/36 and 15/36, respectively) showed significant differences in the percentage of aneuploid cells between 2c and 4c (P = 0.0141) and tetraploid cells (P = 0.0404). In summary, deletions in DNA mismatch repair proteins MSH2 and MLH1 were present both in lentigo maligna and in melanomas and correlated with DNA ploidy-related parameters in malignant melanomas. PMID- 11409566 TI - Tumor necrosis factor alpha (TNFalpha) in human skin: a comparison of different antibodies for immunohistochemistry. AB - Conflicting results have been reported regarding the localization and presence of TNFalpha in normal human skin. To study TNFalpha expression, we tested a panel of antibodies directed against human TNFalpha. First, antibodies were tested for immunoreactivity on cytospots of isolated LPS-stimulated peripheral blood mononuclear cells. Second, antibodies were tested to detect recombinant TNFalpha in Western blots. Some antibodies were found to be unable to detect recombinant TNFalpha in the blots. However, most antibodies were able to bind TNFalpha protein, but they did not bind to other irrelevant proteins that were also present in the blots. Finally, antibodies were tested on cryosections of normal human skin. Antibodies that did not react with TNFalpha in the blots were incubated with TNFalpha before the staining procedure to see whether these antibodies specifically bound TNFalpha. We found that, although all the antibodies bound TNFalpha, there were clear differences in staining patterns. This indicates that these antibodies may recognize distinct epitopes or different forms of TNFalpha. The differences found in this study and those reported previously could be the result of differences in the concentration of antibody used, the staining procedure or specificity of the antibody itself. So, for unambiguous interpretation of data, it is important to know the characteristics of the antibodies used. PMID- 11409567 TI - VEGF expression in skin warts. Relevance to angiogenesis and vasodilation. AB - Verrucae vulgaris (skin warts) are benign proliferative lesions which are generally associated with human papillomavirus type 2 (HPV-2) infection. Vascular endothelial growth factor (VEGF) is an endothelial cell-specific mitogen able to induce angiogenesis and vasodilation. Our previous findings indicate that these two processes take place during the formation of skin warts. The purpose of this study was to determine whether VEGF expression in these lesions was associated with HPV infection, angiogenesis or vasodilation. To this end, paraffin-embedded specimens of skin warts which were either negative for HPV-1, -2, -3 and -4 (HPV ; n = 18), or positive for HPV-2 (HPV+; n = 21) were compared with histologically normal perilesional skin (n = 13). Serial sections were stained with antibodies to von Willebrand Factor (vWF) and to VEGF. Vascularity was quantified by point counting vWF-positive blood vessels. Small and large vessels were quantified separately, using a cut-off value of 50 microm diameter. VEGF expression in the epidermis was estimated by consensus of two independent observers according to three indices: (1) percentage of cells stained, (2) intensity of the staining, and (3) product of area and intensity (final score). Results were analysed by nonparametric tests. Similar levels of VEGF were found in specimens of normal skin, HPV- and HPV+ warts, irrespective of the index used. There was no significant correlation between VEGF expression and vascularity values for either small or large vessels. These results indicate that, on its own, VEGF expression is not associated with angiogenesis, vasodilation or HPV infection in skin warts. The presence of VEGF in normal skin suggests that it may play a role in tissue homeostasis. PMID- 11409568 TI - Altered expression of occludin and tight junction formation in psoriasis. AB - In simple epithelia, tight junctions are well developed and have barrier and fence functions. On the other hand, tight junctions are less developed in stratified epithelia. In the rodent epidermis, only maculae occludentes (i.e. focal strands or spot tight junctions) are observed in the most superficial zone of the granular cell layer. Occludin is an integral membrane protein, and is localized at tight junctions in simple epithelia. In normal epidermis, occludin is expressed at the maculae occludentes in the granular cell layer, indicating that it is associated with keratinocyte differentiation. Thus, we examined occludin expression in psoriasis, in which differentiation of keratinocytes is impaired. In psoriasis, occludin was expressed more broadly in the upper epidermis than in normal epidermis. In addition, immunoelectron microscopy showed occludin to be concentrated on the maculae occludentes in the spinous layer of psoriatic skin. These findings indicate that occludin and the formation of tight junctions are related to the proliferation and differentiation of keratinocytes, and to the pathogenesis of psoriasis. PMID- 11409569 TI - Lipopolysaccharide and cytokines induce nitric oxide synthase and produce nitric oxide in cultured normal human melanocytes. AB - The role of nitric oxide in normal and pathological conditions of human skin is still poorly understood. In this study we have demonstrated by immunobloting the expression of an inducible nitric oxide synthase isoform (iNOS) in cultured normal human melanocytes treated with bacterial lipopolysaccharide, tumor necrosis factor-alpha and interferon-gamma. Nitric oxide was also detected in the culture medium and its formation was abolished upon treatment with N(G)-mono methyl-L-arginine(L-NMMA), a competitive inhibitor of nitric oxide synthase. These results suggest that nitric oxide could led to autodestruction of melanocytes causing skin depigmentation. The therapeutic relevance of nitric oxide synthase inhibitors in treatment of vitiligo was suggested. PMID- 11409570 TI - GM-CSF downmodulates c-kit, Fc(epsilon)RI(alpha) and GM-CSF receptor expression as well as histamine and tryptase levels in cultured human mast cells. AB - GM-CSF is known primarily as a hematopoietic growth factor, but it has also been shown to inhibit mast cell differentiation in vitro. In order elucidate the mechanisms involved, we investigated the effects of GM-CSF in vitro on the differentiation of human leukemic mast cells (HMC-1 cells) and normal cord blood derived mast cells (CBMC) under the influence of SCF, NGF, and fibroblast supernatant (FS). Under all culture conditions, GM-CSF induced a dose- and time dependent reduction in intracellular histamine levels, tryptase activity, and numbers of cells immunoreactive for c-Kit and FcepsilonRIalpha. This effect leveled off between 10-100 ng/ml and after 4 days of culture. There was an associated decrease in mRNA expression for c-kit, FcepsilonRIalpha and tryptase. In contrast, no significant changes in the expression of the NGF receptor TrkA were noted under the same conditions. The GM-CSF receptor was found in HMC-1 cells and CBMC at both the mRNA and protein levels, but its expression decreased during culture with FS, and even more markedly during culture with GM-CSF. GM-CSF thus selectively inhibits in vitro induction and/or upregulation of all major mast cell characteristics in HMC-1 cells and CBMC irrespective of the growth factors present, and a concomitant downregulation of GM-CSF receptors can counteract these effects. GM-CSF may therefore function as a regulatory factor in mast cell growth and differentiation under normal and pathological conditions. PMID- 11409571 TI - Expression profiles of cell-cell and cell-matrix junction proteins in developing human epidermis. AB - The spatial distribution of a panel of cell junction proteins was studied in developing human epidermis by confocal laser scanning microscopy. The results demonstrated that many of the cell junction proteins were expressed in early two layered embryonic epidermis, but their subcellular distribution displayed marked changes during development. Specifically, desmosomal proteins, desmoplakin, desmocollin and desmoglein, adherens junction components, E-cadherin, alpha catenin and vinculin, and an actin-binding protein alpha-actinin were expressed as early as 8 weeks of estimated gestational age (EGA). Type IV collagen and beta1 and beta4 integrins were also present. At this early developmental stage, the epidermis is known to comprise two layers of cells, the basal layer and the peridermal layer. In addition to being present in cell-cell contacts, desmosomal antigens and E-cadherin were unexpectedly localized to the basal aspect of basal cells in samples at 8 weeks. On the other hand, talin, which in adult skin is localized to the dermal-epidermal junction, could not be detected until 12 weeks. These results suggest that the separation of cell membranes to the basal and apicolateral compartments does not occur before the maturation of the basement membrane zone. At 8 weeks EGA, gap junction antigen connexin 43 was expressed in scarce spots in cell-cell contacts of basal cells. In samples of 11-21 weeks EGA, the density of desmosomal and adherens junction markers as well as connexin 43 increased in cell-cell junctions, together with the appearance of the intermediate cell layer and beginning of stratification of the epidermis. PMID- 11409572 TI - In a skin equivalent HaCaT cells have a preserved capacity to receive melanosomes but melanocytes do not remain in the basal location. PMID- 11409573 TI - Appropriate technology for the administration of oxygen to children at district hospitals in developing countries. PMID- 11409574 TI - Epidemiology of hypoxaemia in children with acute lower respiratory infection. AB - OBJECTIVES: To determine the prevalence of hypoxaemia in children aged under 5 years suffering acute lower respiratory infections (ALRI), the risk factors for hypoxaemia in children under 5 years of age with ALRI, and the association of hypoxaemia with an increased risk of dying in children of the same age. DESIGN: Systematic review of the published literature. SETTINGS: Out-patient clinics, emergency departments and hospitalisation wards in 23 health centres from 10 countries. SUBJECTS: Cohort studies reporting the frequency of hypoxaemia in children under 5 years of age with ALRI, and the association between hypoxaemia and the risk of dying. MAIN OUTCOME MEASURES: Prevalence of hypoxaemia measured in children with ARI and relative risks for the association between the severity of illness and the frequency of hypoxaemia, and between hypoxaemia and the risk of dying. RESULTS: Seventeen published studies were found that included 4,021 children under 5 with acute respiratory infections (ARI) and reported the prevalence of hypoxaemia. Out-patient children and those with a clinical diagnosis of upper ARI had a low risk of hypoxaemia (pooled estimate of 6% to 9%). The prevalence increased to 31% and to 43% in patients in emergency departments and in cases with clinical pneumonia, respectively, and it was even higher among hospitalised children (47%) and in those with radiographically confirmed pneumonia (72%). The cumulated data also suggest that hypoxaemia is more frequent in children living at high altitude. Three papers reported an association between hypoxaemia and death, with relative risks varying between 1.4 and 4.6. Papers describing predictors of hypoxaemia have focused on clinical signs for detecting hypoxaemia rather than on identifying risk factors for developing this complication. CONCLUSION: Hypoxaemia is a common and potentially lethal complication of ALRI in children under 5, particularly among those with severe disease and those living at high altitude. Given the observed high prevalence of hypoxaemia and its likely association with increased mortality, efforts should be made to improve the detection of hypoxaemia and to provide oxygen earlier to more children with severe ALRI. PMID- 11409575 TI - Clinical signs of hypoxaemia in children with acute lower respiratory infection: indicators of oxygen therapy. AB - Hypoxaemia is a common complication of acute lower respiratory tract infections in children. In most developing countries, where the majority of deaths from pneumonia occur, facilities for early detection of hypoxaemia are lacking and oxygen is in short supply. This review examines the usefulness of different clinical signs and symptoms in the prediction of hypoxaemia associated with acute respiratory infections in children. Several respiratory signs were found to be associated with hypoxaemia. These include very fast breathing (with a respiratory rate of more than 60 or 70 breaths per minute), cyanosis, grunting, nasal flaring, chest retractions, head nodding and auscultatory signs, as well as signs of general depression of the child, such as inability to feed or lethargy. The sensitivity and specificity of these signs, as described in the reviewed studies, is presented, and combination rules are discussed. Through appropriate combination of several physical signs, which can be used by peripheral health workers and be taught to mothers, it is possible to predict hypoxaemia in children with acute respiratory tract infections with reasonable accuracy. PMID- 11409576 TI - Hypoxaemia in children with severe pneumonia in Papua New Guinea. AB - OBJECTIVES: To investigate the severity and duration of hypoxaemia in 703 children with severe or very severe pneumonia presenting to Goroka Hospital in the Papua New Guinea highlands; to study the predictive value of clinical signs for the severity of hypoxaemia, the predictive value of transcutaneous oxygen saturation (SpO2) and other variables for mortality. DESIGN: Prospective evaluation of children with severe or very severe pneumonia. SpO2 was measured at the time of presentation and every day until hypoxaemia resolved. Children with a SpO2 less than 85% received supplemental oxygen. By comparing with a retrospective control group for whom oxygen administration was guided by clinical signs, we evaluated whether there was a survival advantage from using a protocol for the administration of oxygen based on pulse oximetry. We determined normal values for oxygen saturation in children living in the highlands. RESULTS: In 151 well, normal highland children, the mean SpO2 was 95.7% (SD 2.7%). The median SpO2 among children with severe or very severe pneumonia was 70% (56-77); 376 (53.5%) had moderate hypoxaemia (SpO2 70-84%); 202 (28.7%) had severe hypoxaemia (SpO2 50-69%); and 125 (17.8%) had very severe hypoxaemia (SpO2 < 50%). Longer duration of cough or the presence of hepatomegaly or cyanosis predicted more severe degrees of hypoxaemia. After 10, 20 and 30 days from the beginning of treatment, respectively 102 (14.5%), 38 (5.4%) and 19 (2.7%) of children had persistent hypoxaemia; 46 children (6.5%) died. Predictors of death were low SpO2 on presentation, severe malnutrition, measles and history of cough for more than 7 days. The mortality risk ratio between the 703 children managed whose oxygen administration was guided by the use of pulse oximetry and the retrospective control group who received supplemental oxygen based on clinical signs was 0.65 (95%CI 0.41-1.02, two-sided Fisher's exact test, P = 0.07). CONCLUSION: There is a need to increase the availability of supplemental oxygen in smaller health facilities in developing countries, and to train health workers to recognise the clinical signs and risk factors for hypoxaemia. In moderate sized hospitals a protocol for the administration of oxygen based on pulse oximetry may improve survival. PMID- 11409577 TI - Oxygen concentrators and cylinders. AB - A comparison is made between oxygen cylinders and oxygen concentrators as sources for clinical use. Oxygen cylinders are in widespread use, but costs and logistic factors favour the use of concentrators in many developing country situations, especially where cylinder supplies fail to penetrate. PMID- 11409578 TI - Oxygen supply in rural africa: a personal experience. AB - Oxygen is one of the essential medical supplies in any hospital setting. However, in some rural African settings without regular electricity and with logistical problems in the transport of oxygen cylinders, the delivery of oxygen can be difficult. This paper compares the costs incurred using oxygen cylinders with a solar operated oxygen concentrator in a rural Gambian setting. The paper shows that a solar operated system has a high capital investment initially, but that running costs and maintenance are minimal. The system becomes cost effective if a rural hospital needs more than 6 treatment days (1 l/min) of oxygen per month and can be considered in a setting where 6 hours of sunlight per day can be guaranteed. PMID- 11409579 TI - Oxygen delivery to children with hypoxaemia in small hospitals in developing countries. AB - Oxygen administration is one of the most important modalities of therapy for a patient with hypoxaemia to prevent death. This review summarises the methods of oxygen delivery applicable in small hospitals in developing countries, and evaluates information about their safety and efficacy. The following criteria were considered: cost and availability, efficiency/oxygen concentration achieved, tolerability/comfort of the method, requirement of humidification, demand for nursing care, and safety of the method and complications. In summary, it is concluded that all low-flow methods, i.e., nasopharyngeal catheters, nasal catheters and prongs, are effective in the oxygenation of sick children with severe pneumonia or bronchiolitis. Nasal prongs are the safest method of oxygen delivery, but nasopharyngeal catheters and nasal catheters are more easily available and less expensive. However, if they are used, they need close supervision to avoid serious complications. Nasal prongs are the method of choice for oxygen delivery in small hospitals in developing countries. PMID- 11409580 TI - The early bactericidal activity of amikacin in pulmonary tuberculosis. AB - SETTING: Stellenbosch University, a tertiary care hospital in Cape Town, South Africa. OBJECTIVE: To determine the early bactericidal activity (EBA) of amikacin in dosages of 5 mg/kg, 10 mg/kg and 15 mg/kg body weight in comparison to that of isoniazid 6 mg/kg body weight or no drug. DESIGN: An open, randomised trial. PATIENTS: Patients with previously untreated, sputum smear-positive pulmonary tuberculosis. INTERVENTION: Patients received amikacin 5 mg/kg (12 patients), 10 mg/kg (13 patients) or 15 mg/kg (15 patients), isoniazid 6 mg/kg (9 patients) or no drug (10 patients). RESULTS: The rate of decrease in log viable colony forming units of Mycobacterium tuberculosis per ml of sputum per day during the first 2 days of treatment with amikacin 5 mg/kg, 10 mg/kg and 15 mg/kg was 0.041 (SD 0.100), 0.045 (SD 0.144) and 0.052 (SD 0.096), respectively, 0.515 (SD 0.173) in the patients receiving isoniazid 6 mg/kg, and 0.041 (SD 0.113) in those receiving no drug. The EBA found in patients receiving amikacin did not differ significantly from that of the no drug group. However, as the EBA in the no drug group was the highest ever encountered at Stellenbosch University, the mean in patients receiving drug was tested against 0 and found to differ significantly (P = 0.03), suggesting minimal activity. Mean amikacin serum concentrations 1 hour after intramuscular drug administration were 13.5 microg/ml, 26.7 microg/ml and 39.2 microg/ml in the patients receiving 5 mg, 10 mg and 15 mg per kg body weight, respectively. CONCLUSION: Despite serum concentrations well in excess of the minimal inhibitory concentration of 2-4 microg/ml, the EBA of amikacin in patients with smear-positive pulmonary tuberculosis was only just detectable. PMID- 11409581 TI - The relationship between the in vitro drug susceptibility of opportunist mycobacteria and their in vivo response to treatment. AB - SETTING: It is generally accepted that qualitative drug susceptibility tests established and validated for Mycobacterium tuberculosis are not applicable to opportunist (non-tuberculous) mycobacteria. Previous studies have shown that in vitro antimicrobial susceptibilities for opportunist mycobacteria, performed by the method of modal resistance (MR), correlate poorly with clinical response. Minimum inhibitory concentration (MIC) determination may provide better correlation with predicted clinical response than the conventional MR results. OBJECTIVE: To determine the relationship between quantitative in vitro sensitivity results for opportunist mycobacteria and their in vivo response to treatment. DESIGN: MICs were performed radiometrically with the Bactec TB-460 system; 35 M. avium complex isolates, 29 isolates of M. malmoense and 16 isolates of M. xenopi were tested. RESULTS: Susceptibility results were analysed in comparison with therapeutic outcome by Fisher's exact probability test. Only one significant association was found; in vitro resistance to ethambutol correlated with treatment failure for M. malmoense infections (P = 0.027). There were no other significant correlations between in vitro results and treatment outcome. CONCLUSION: Prediction of treatment outcome from in vitro susceptibility tests continues to be a problem in infections with opportunist mycobacteria. PMID- 11409582 TI - Increased resistance to ciprofloxacin and ofloxacin in multidrug-resistant mycobacterium tuberculosis isolates from patients seen at a tertiary hospital in the Philippines. AB - SETTING: A hospital-based study at the Makati Medical Center, Makati City, Philippines, a hyperendemic area for tuberculosis (TB). OBJECTIVE: To determine the susceptibility of Mycobacterium tuberculosis to ciprofloxacin and ofloxacin. DESIGN: Retrospective analysis of drug susceptibility tests (DST) of M. tuberculosis isolated from 1995-2000. RESULTS: Resistance to ciprofloxacin was 26.8%, ofloxacin 35.3%, and multidrug resistance (MDR) was 17.2%. Of the MDR strains, 51.4% were resistant to ciprofloxacin and ofloxacin. Acquired resistance was significantly higher for all first-line drugs and for ciprofloxacin, but not for ofloxacin. A significant increase in resistance to ciprofloxacin and ofloxacin was noted compared to 1989-1994, while resistance to the firstline drugs was not significantly different. CONCLUSION: Ciprofloxacin and ofloxacin are now a significantly less effective alternative therapy in tuberculosis, particularly MDR-TB, due to a selection pressure from their widespread use in the treatment of TB and possibly other infections in the community, which is hyperendemic for tuberculosis. PMID- 11409583 TI - Familial outbreak of disseminated multidrug-resistant tuberculosis and meningitis. AB - Rapidly progressive multidrug-resistant tuberculosis (MDR-TB) is well documented in human immunodeficiency virus (HIV) positive subjects, but it is not fully recognised in HIV-negative subjects in the familial environment. We report three cases of MDR-TB in three young HIV-negative subjects from the same family. All the patients showed signs of meningitis during the course of their disease, and in two cases a resistant strain of Mycobacterium tuberculosis was isolated in cerebrospinal fluid. Two of the three subjects died from neurological complications; the other was successful treated utilising both systemic and intrathecal therapy for tuberculous meningitis. By a retrospective analysis of DNA obtained from Lowenstein-Jensen cultures, the strains were confirmed as M. tuberculosis resistant to rifampicin and isoniazid, and were closely related in the two cases where specimens were available for analysis. The resistance was acquired in two patients initially infected with a susceptible strain; in the other patient, the resistance was present on the first sensitivity test for which results were available. This report demonstrates the high risk of fatality from MDR-TB for HIV-negative subjects in the absence of reliable early diagnostic and preventive tools. It also reinforces the concept that genetic susceptibility to M. tuberculosis may be an important factor in the clinical presentation and outcome of MDR-TB. PMID- 11409584 TI - Efficacy and safety of sparfloxacin in combination with kanamycin and ethionamide in multidrug-resistant pulmonary tuberculosis patients: preliminary results. AB - SETTING: Department of tuberculosis and chest diseases of a tertiary referral tuberculosis institute in New Delhi, India. OBJECTIVE: To study the efficacy and safety of sparfloxacin, in combination with kanamycin (for the initial 3-4 months) and ethionamide, in multidrug-resistant (MDR) pulmonary tuberculosis patients. DESIGN: Prospective, uncontrolled study of nine patients with pulmonary tuberculosis who had received adequate anti-tuberculosis treatment with first line drugs, including supervised category II treatment regimen as per World Health Organization guidelines for 5 months, and were still sputum smear acid fast bacilli positive. It was planned to give them kanamycin (initial 3-4 months), ethionamide and sparfloxacin for 2 years. RESULTS: All nine patients achieved sputum conversion within 6 months. Seven patients converted within 3.5 months, two of these within 1 month. All patients reported improvement in clinical symptoms, and chest X-ray improved in seven patients. Four patients developed mild to moderate phototoxicity. Eight patients have completed treatment for an average of 19 months (range 15-24 months), and are still under follow-up. One patient defaulted after 7 months of treatment. CONCLUSIONS: In MDR-TB patients, sparfloxacin, along with other anti-tuberculosis drugs, appears to be effective and safe. Mild to moderate phototoxicity is common. However, the long term results, including relapses, are still awaited. PMID- 11409585 TI - A population-based case-control study of the safety of oral anti-tuberculosis drug treatment during pregnancy. AB - OBJECTIVE: To study the human teratogenic potential of isoniazid and other anti tuberculosis drug treatment during pregnancy. DESIGN AND SETTING: Cases from a large population-based dataset at the Hungarian Case-Control Surveillance of Congenital Abnormalities, and controls from the National Birth Registry, between 1980 and 1996. Information on all oral anti-tuberculosis drug treatments during pregnancy was medically recorded. STUDY PARTICIPANTS: Women who had newborns or fetuses with congenital abnormalities (case group), and women who had babies with no congenital abnormality (control group). MAIN OUTCOME MEASURES: Congenital abnormalities in newborn infants and fetuses diagnosed prenatally during the second and third trimesters, and postnatally from birth to the age of one year. RESULTS: Of 38,151 controls, 29 (0.08%) were exposed to anti-tuberculosis drug treatment during pregnancy; the corresponding figures for cases were 22,865 and 11 (0.05%). The prevalence odds ratio was 0.6 (95%CI 0.3-1.3). Analysis of isoniazid and other oral antituberculosis drug use during the second and third months of gestation, i.e., in the critical period for most major congenital abnormalities, in case-control pairs did not indicate a teratogenic effect of these drugs in any group with congenital abnormality. CONCLUSION: Maternal exposure to oral anti-tuberculosis drugs during pregnancy did not show a detectable teratogenic risk to the fetus; however, the number of pregnant women who were treated with these drugs during the critical period of most major congenital abnormalities was limited (six cases vs. 21 controls). PMID- 11409586 TI - Specificity of insertion sequence-based PCR assays for mycobacterium tuberculosis complex. AB - OBJECTIVE: To determine the specificity of different insertion sequence-targeted polymerase chain reaction (PCR) tests for Mycobacterium tuberculosis complex. DESIGN: One M. bovis BCG strain, two M. tuberculosis strains and ten species of mycobacteria other than tuberculosis (MOTT) were tested by three PCR assays based on the repetitive elements IS6110, IS1081 and IS990 under variable amplification conditions (different temperatures of primer annealing and numbers of reaction cycles). RESULTS: DNA amplifications based on the three insertion sequences yielded fragments of expected sizes only in DNA from M. tuberculosis complex strains when the tests were conducted at high stringency (65 degrees C). At the annealing temperature of 60 degrees C the PCR assay with IS6110-specific primers yielded a 245 bp fragment also in nine MOTT strains tested. This could result from previously reported homology between non-tuberculous mycobacteria and a central region of IS6110. Amplification assays based on IS1081 and IS990 gave false-positive results in some MOTT isolates only under very low stringency (55 degrees C), which could be due to non-specific priming of the target DNA at that temperature. CONCLUSION: Repetitive elements IS1081 and IS990 may represent a more reliable alternative to the more widely used IS6110 PCR target for tuberculosis diagnosis. PMID- 11409587 TI - A case series: initial outcome of persons with multidrug-resistant tuberculosis after treatment with the WHO standard retreatment regimen in Ho Chi Minh City, Vietnam. AB - Few data address the outcomes of patients who have multidrug-resistant tuberculosis (MDR-TB), defined as resistance to at least isoniazid and rifampin, and who receive a standard World Health Organization (WHO) recommended retreatment regimen after relapse or failure with initial treatment. In this case series, we examined treatment outcomes of a convenience sample of 42 relapse or failure patients who had documented MDR-TB and who had received a standard WHO retreatment regimen (2SHRZE/1HRZE/5H3R3E3). One patient died of tuberculosis in the last month of treatment; the remaining 41 patients completed retreatment. Of the 42, 14 (33%) were sputum smear-negative on completion of therapy. The proportion of patients cured of MDR-TB with the WHO retreatment regimen was similar to historic outcomes when no chemotherapy for TB was given. PMID- 11409588 TI - Quantitative bacillary response to treatment in Mycobacterium tuberculosis infected and M. africanum infected adults with pulmonary tuberculosis. AB - Data regarding possible differences in microbiological response to therapy of disease caused by Mycobacterium tuberculosis and M. africanum are limited. Presenting clinical characteristics and sputum bacillary load during standard short-course chemotherapy in patients with newly-diagnosed pulmonary tuberculosis due to M. tuberculosis (n = 7) and M. africanum (n = 6) were compared. Changes in sputum bacillary load were measured using quantitative acid-fast bacilli smears, colony forming unit assay, and time until positive culture in the BACTEC radiometric system. Presentation and response to short course chemotherapy were comparable between patients infected with M. tuberculosis and those infected with M. africanum. PMID- 11409589 TI - Tuberculous infection in student nurses in Madrid, Spain. AB - To determine the incidence of tuberculous infection in student nurses we performed a 3-year study in our hospital. Before training was initiated, 18.75% of males and 5.7% of females were Mantoux-positive (P = 0.09). During the following two years, 9.2% of the previously Mantoux-negative students became positive. No differences were found between males and females. We conclude that student nurses must be considered at risk for tuberculous infection in our hospital, and that stricter isolation procedures for tuberculous patients must be implemented. PMID- 11409590 TI - Serological diagnosis of tuberculosis using IgA detection against the mycobacterial kp-90 antigen. PMID- 11409591 TI - Evidence for the transmission of tuberculosis in a prison in Nakhon Ratchasima, Thailand. PMID- 11409592 TI - High-resolution computed tomography classification of lung fibrosis for patients with asbestos-related disease. AB - OBJECTIVES: This study tested a new high-resolution computed tomography (HRCT) scoring method for asbestos-induced parenchymal changes in the lung. METHODS: HRCT scans of 602 asbestos-exposed workers and 49 referents were reviewed by 3 radiologists. Structured forms were filled out for a semiquantitative HRCT fibrosis score based on several specified parenchymal abnormalities scored separately. Observer agreement was studied with the use of the quadratic-weighted kappa (kappaqw). The HRCT fibrosis score (from 0 to V with definitions and index images given retrospectively) was compared with the radiographic classification of the International Labour Office (ILO) for the same patients. Receiver operating characteristic (ROC) curves were computed to compare the tests for diagnosing asbestosis. RESULTS: Good inter- and intraobserver agreements were achieved (kappaqw = 0.64 and 0.72, respectively) as regards the HRCT fibrosis score. All the specified computed tomography findings explained 86% of the variance in the HRCT fibrosis score. Age and occupational group were significant predictors of fibrosis. The area under the ROC curve was significantly greater for the HRCT fibrosis score (0.89) than for the ILO radiographic classification (0.76). The sensitivity (70%) and specificity (91%) of the HRCT fibrosis score (classes I/II-V representing asbestosis) were better than those of the classification published by the International Labour Office (51% and 89%, respectively, score > or = 1/0 representing asbestosis). CONCLUSIONS: The examined HRCT scoring method proved to be a simple, reliable, and reproducible method for classifying lung fibrosis and diagnosing asbestosis also in large populations with occupational disease, and it would be possible to use it as a part of an international classification. PMID- 11409593 TI - Cohort cancer incidence among pulp and paper mill workers in British Columbia. AB - OBJECTIVES: A study was conducted to investigate cancer risks in a cohort of pulp and paper workers. METHODS: All male workers with > or =1 years of employment in 14 pulp and paper mills in 1950-1992 were studied. Standardized incidence ratios (SIR) were used to compare the cancer incidence of the cohort with that of the Canadian male population. Record linkage with the National Cancer Registry was performed using the generalized iterative record linkage method. RESULTS: Altogether 1756 cancer cases were observed in the entire cohort. For > or =15 years of work, the entire cohort had significantly increased SIR values for pleural and prostate cancer and skin melanoma; there was also a significantly increased risk for skin melanoma among workers in the kraft process only, rectal cancer among workers in the sulfite process only, and stomach and prostate cancer and all leukemias combined among workers in both the kraft and sulfite processes. A separate analysis comparing workers in pulping and papermaking with those in the pulping process only did not reveal any difference in cancer risk and hence did not modify the results. The SIR values for skin melanoma were not significantly increased in a comparison using the British Columbia male population. Nine of 10 pleural cancers were mesotheliomas, which likely reflect past asbestos exposure. CONCLUSIONS: The results suggest that long-term work in the pulp and paper industry is associated with excess risks of prostate and stomach cancers and all leukemias for work in both kraft and sulfite processes and of rectal cancer for work in the sulfite process only. PMID- 11409594 TI - Chromosome aberrations in lymphocytes of persons exposed to an earthquake in Armenia. AB - OBJECTIVE: This study attempted to determine the level of chromosome aberrations in lymphocytes of victims of the 1988 earthquake in Armenia. METHODS: Chromosome aberrations were measured in blood samples taken from 41 victims of the earthquake that hit Armenia in 1988 and in samples of 47 reference blood donors. The victims suffered from severe psychoemotional stress but were otherwise healthy. All the samples were taken 2 to 3 weeks after the earthquake. All the subjects were lifetime nonsmokers. The cells were scored blind as to the exposure status. RESULTS: The subjects exposed to the earthquake had a higher proportion of cells with chromosome aberrations [3.1 (SD 2.1)%] than the referents [1.7 (SD 1.3)%, P-value for the difference 0.0009]. The difference persisted when the values were adjusted for age and gender [relative risk (RR) 1.9, 95% confidence interval (95% CI) 1.4-2.5]. The difference was present for double breaks (RR 4.1, 95% CI 2.6-6.4), but not for single breaks (RR 1.1, 95% CI 0.8-1.7). The exposed subjects also had a lower percentage of cells with 46 chromosomes (P=0.03) than the referents. CONCLUSIONS: This study suggests an increase in chromosome aberrations in the lymphocytes of victims of a severe earthquake as compared with the levels of referents. If not due to bias or confounding, the difference may reflect the effect of either environmental exposures related to the earthquake or severe psychogenic stress. The levels of chromosome aberrations found among the earthquake victims in this study are comparable with those found in prospective studies of long-term cancer risk. PMID- 11409595 TI - Group-based measurement strategies in exposure assessment explored by bootstrapping. AB - OBJECTIVES: The precision of mean exposure to pushing was examined in 2 occupational groups using various combinations of the number of workers and measurements per worker. METHODS: The frequency and duration of pushing of the 2 occupational groups was assessed using onsite observation. All data were divided into successive periods of 30 minutes of observation. The precision of the group mean exposure to pushing was expressed by 90% confidence intervals obtained by bootstrapping. The effect on the confidence interval of varying numbers of workers and numbers of periods per worker was examined. RESULTS: For both occupational groups there was little precision to be gained when >10 workers were observed. Within the maximum number of workers used in the bootstrap simulations, it appeared that, beyond 10 workers, the confidence intervals decreased by <5% for every worker that was added, when each worker was observed at least 8 periods of 30 minutes. If workers were observed exactly 4 periods of 30 minutes per worker, an additional 4 workers were required to compensate for the loss of precision. An unbalanced strategy with approximately 8 periods of 30 minutes per worker hardly decreased the precision of the group mean, however. CONCLUSIONS: The precision of the group-based mean exposure to pushing is influenced by the number of workers observed and by the number of repeated measurements per worker. In the planning of measurement strategies, it is advisable to account for possible sources of variance in advance and to assess the exposure variability. PMID- 11409596 TI - Total plasma protein adducts of allergenic hexahydrophthalic and methylhexahydrophthalic anhydrides as biomarkers of long-term exposure. AB - OBJECTIVES: The aim of this study was to evaluate the applicability of total plasma protein adducts (TPPA) of 2 sensitizing low-molecular-weight allergens, hexahydrophthalic anhydride (HHPA) and methylhexahydrophthalic anhydride (MHHPA), as biomarkers of long-term exposure. METHODS: Urine samples from occupationally exposed workers were analyzed for the levels of urinary metabolites of HHPA and MHHPA, and the levels were used as the index of exposure. In addition, blood samples were obtained from the same persons, and the levels of TPPA were determined. Reversed solid phase extraction, derivatization using pentafluorobenzyl bromide, and gas chromatography-mass spectrometry analysis in the negative ion chemical ionization mode were used to quantify the exposure. To assess the suitability of TPPA as a biomarker of exposure to the anhydrides, the TPPA levels were correlated to urinary metabolite levels and hemoglobin (Hb) adducts. The toxicokinetics of TPPA were also studied to determine the elimination half-time of the adducts. RESULTS: The levels of TPPA correlated exceptionally well with the metabolite levels in the urine sampled repeatedly, giving r=0.97 for HHPA and r=0.92 for MHHPA. The TPPA of HHPA correlated highly with the Hb adducts with r=0.86. There were also good correlations between single urinary determinations and the TPPA levels (r(s)=0.71 and 0.81, respectively, for HHPA and MHHPA). The in vivo decay of TPPA gave an elimination half-time of 22 days for HHPA and 24 days for MHHPA. CONCLUSIONS: TPPA levels of HHPA and MHHPA are excellent biomarkers of long-term exposure to anhydrides. PMID- 11409597 TI - Effects of electric field reduction in visual display units on skin symptoms. AB - OBJECTIVES: This study investigated the facial skin complaints of office workers before and after the static electric fields of a visual display unit were reduced. METHODS: On the basis of a screening survey of 4556 office workers in 11 companies, 120 of 227 subjects reporting facial skin complaints were randomly selected to this double blind intervention study. Antistatic measures were used to reduce the static electric fields of the visual display unit in the intervention group but not in the control group, which worked with a visual display unit resembling that of the intervention group. Electric fields, dust concentration, health complaints, and psychological behavior tests were recorded before and after the intervention. RESULTS: The intervention group reported statistically significantly fewer facial skin complaints than the control group. In the intervention group, among those with an office dust concentration of >58 microg/m3, a median reduction of 1.5 skin index points (scale 0-8) was achieved, whereas there was no change in the control group. In the regression model "group category" was still a significant variable after control for background factors. In addition, further linear regression analyses indicated that several static electric field parameters were predictors of the skin complaint reduction. CONCLUSIONS: This field trial indicates that removing static electric fields from visual display units can probably help reduce the facial skin complaints of workers in offices with high dust concentrations. PMID- 11409598 TI - Association between job stress and depression among Japanese employees threatened by job loss in a comparison between two complementary job-stress models. AB - OBJECTIVES: This study compared the separate effects produced by two complementary stress models--the job demand-control model and the effort-reward imbalance model--on depression among employees threatened by job loss. METHODS: A cross-sectional analysis was conducted to examine these associations among 190 male and female employees who responded to a self-administered questionnaire in a small Japanese plant with economic hardship. The employees were engaged in 2 job types--direct assembly line and indirect supportive tasks--and the latter was threatened by job loss because of downsizing. Independent variables were measured by the Japanese versions of Karasek's demand-control questionnaire and Siegrist's effort-reward imbalance questionnaire. Depression was assessed by the Center for Epidemiologic Studies Depression Scale. RESULTS: The employees with indirect supportive tasks (target for downsizing) were more likely to have depressive symptoms than direct assembly-line workers. Job strain, a combination of high demand and low control at work, was more frequent among the latter, while the combination of high effort and low reward was more frequent among the former. After adjustment for work environment factors, low control [odds ratio (OR) 4.7], effort reward imbalance (OR 4.1), and overcommitment (the person characteristic included in the effort-reward imbalance model) (OR 2.6) were independently related to depression. There is some indication that these effects were particularly strong in the subgroup suffering from potential job loss. CONCLUSIONS: This study confirms that the 2 job stress models identify different aspects of stressful job conditions. Moreover, effort-reward imbalance and low control at work are both associated with symptoms of depression. PMID- 11409599 TI - Association between pleural plaques and coronary heart disease. AB - OBJECTIVE: The aim of this study was to verify a clinical impression that patients with coronary heart disease disproportionately frequently have calcified pleural plaques. METHODS: Chest X-rays were collected from 148 patients referred consecutively to the Helsinki University Central Hospital for coronary angiography and from 100 consecutive lung cancer patients seen at the same hospital. The radiographs were analyzed for the presence of calcified pleural plaques according to the classification the International Labour Office. A generalized linear model with binomial distribution and log link was used to estimate the relative risks and their 95% confidence intervals (95% CI). RESULTS: The prevalence of calcified pleural plaques was 35% for the coronary patients and 19% for the lung cancer patients. Calcified pleural plaques were more common among the men than the women, and the risk increased with age. The relative risk of calcified pleural plaques, adjusted for age and gender, was 2.19 (95% CI 1.44 3.32) for the coronary patients as compared with the lung cancer patients. CONCLUSIONS: Further studies with better information on past exposure to asbestos and other potential risk factors are warranted to confirm the observations and to examine whether the association between coronary heart disease and calcified pleural plaques is related to an etiologic or an individual susceptibility factor common to both of these conditions. PMID- 11409600 TI - Shift work and cardiovascular disease--from etiologic studies to prevention through scheduling. PMID- 11409601 TI - Intervention in shift scheduling and changes in biomarkers of heart disease in hospital wards. AB - OBJECTIVES: The effect of introducing regularity, few consecutive night shifts, more weekends off, and only 2 different types of shifts (day-evening or day night) into shift scheduling on biomarkers of heart disease was studied. METHODS: Ergonomic shift criteria were introduced in a quasi-experimental controlled intervention in 4 hospital wards. Six wards participated as controls. Altogether 101 nurses and nurses' aides were followed for 6 months with measurements of cholesterol and triglycerides. The intervention led to more regular schedules and more staff having 2 shifts in 2 of the intervention wards 1 year after the intervention. The schedules among the controls became less regular and less predictable. The number of consecutive night shifts remained unchanged. RESULTS: After 6 months the high-density lipoprotein (HDL) cholesterol level had increased in the intervention group, and the total cholesterol and low-density lipoprotein (LDL) cholesterol levels and the total:HDL cholesterol ratio had decreased. Regardless of the intervention, changes in regularity were associated with the triglyceride and HDL cholesterol levels and also with the total:HDL cholesterol ratio. More ergonomic changes were associated with lower LDL cholesterol levels, a lower total:HDL cholesterol ratio, and higher HDL cholesterol levels. CONCLUSIONS: Increased ergonomic scheduling was possible. Lipids and lipoproteins changed as predicted, both when the changes were assessed in respect to the changes in schedules that resulted from the intervention and the changes that occurred regardless of the intervention. The study suggests that scheduling based on ergonomic criteria is a possible means for reducing the risk of heart disease among shift workers. PMID- 11409602 TI - Work environment of Danish shift and day workers. AB - OBJECTIVES: Both shift work and other work environment factors have been shown to be related to heart disease. This study examined whether shift work is associated with other work environment factors related to heart disease in a random sample of the population. If so, shift work could be acting as a proxy for work environment differences. METHODS: Data on 5940 employees in the Danish Work Environment Cohort Study from 1990 were reanalyzed. The information included work schedules [permanent day work, irregular workhours (including morning work), 2 shift or fixed evening and 3-shift or fixed night], length of workweek, physical factors (noise, heat, dust, passive smoking, walking, standing and monotonous repetitive tasks), and psychosocial factors (including demands and control dimensions, social support, conflicts and job insecurity). RESULTS: At least 1 group of shift workers had a higher prevalence of nearly every unfavorable work environment factor investigated. Exceptions were dust exposure and quantitative demands. Especially conflicts at work and low decision latitude were higher among all the groups of shift workers, and all-day walking or standing work and part time jobs were more often found among female shift workers. The 3 different shiftwork groups were exposed to different parts of the work environment, and also men and women in shift work differed in relation to the work environment. Age and social class influenced the relationship, but not in any particular pattern. CONCLUSIONS: In a heterogenous population shift work was found to be associated with other work environment factors suspected to cause heart disease. PMID- 11409603 TI - Relationships among types of speech intelligibility in pediatric users of cochlear implants. AB - Twenty pediatric users of cochlear implants were administered three tests of speech intelligibility: (1) a test of contrast perception intelligibility, (2) a test of contrast production intelligibility, and (3) a test of production sentence intelligibility. Sixty adults with normal hearing served as listener judges for the two speech production tasks, and percent correct scores were generated for each of the three tasks. Correlational analyses showed significant correlations among overall scores for the three tasks. However, scores for individual feature classes from the contrast perception task were not correlated with their corresponding contrast production feature class scores, and only some of the feature class scores were correlated significantly with sentence intelligibility. We conclude that although these three types of intelligibility are related at a gross level, relationships are more tenuous at finer levels of analysis, suggesting that the separate skills may need to be addressed separately in remediation. EDUCATIONAL OBJECTIVES: As a result of this activity, the participant will be able to differentiate various methods for assessing speech intelligibility and describe the relationships among different types of speech intelligibility in pediatric users of cochlear implants. PMID- 11409604 TI - Third person pronoun errors by children with and without language impairment. AB - Research findings have been mixed about pronoun case problems in the language learning profile of children with specific language impairment (SLI). This study (N= 36) extended previous findings and located a number of error patterns using detailed error analyses. Results indicated that the children with expressive SLI produced more errors with third person singular (3Psg) pronouns than did their age-level peers, but they did not make more errors than their MLU-matched peers. Error patterns were similar in the children with SLI and their language-level peers. The most frequent type of error was the substitution of the objective case for the nominative case. More errors were made on the feminine pronoun, she, than on the masculine pronoun, he. Implications for theories and clinical practice were explored. EDUCATIONAL OBJECTIVES: As a result of this activity, the reader will (1) learn how children with SLI compare to their peers in producing third person pronouns, (2) learn the most common types of pronoun errors made by the children matched for mean length of utterance (MLU), and (3) evaluate how the findings relate to two current theories: one from typical language development and one from the area of SLI. PMID- 11409605 TI - Gender effects in auditory brainstem responses to air- and bone-conducted clicks in neonates. AB - Examinations of gender differences in auditory brainstem response (ABR) wave V latencies and thresholds to air- and bone-conducted clicks were undertaken with neonates. Two hundred and two full-term neonates participated (i.e., 103 males and 99 females). Wave V latency measures for air- and bone-conducted click stimuli of 30, 45, and 60 dB nHL and 15 and 30 dB nHL, respectively, and thresholds to air- and bone-conducted clicks were determined. Female newborns displayed statistically significant shorter wave V latencies than male newborns for air-conducted click stimuli (i.e., approximately 0.2-0.3 ms; P=.0016). There were no significant gender differences in wave V latencies to bone-conducted click stimuli (P=.11). With respect to ABR thresholds, no statistically significant differences were observed for either air-conducted clicks (P=.054) or bone-conducted clicks (P=.18). EDUCATIONAL OBJECTIVES: As a result of this activity, the participant will be able to (1) describe gender differences in ABR wave V latencies and thresholds to air- and bone-conducted clicks with neonates and (2) summarize possible explanations for observed gender differences in ABR wave V latencies and thresholds to air- and bone-conducted clicks with neonates. PMID- 11409606 TI - Family education seminars and social functioning of adults with chronic aphasia. AB - Aphasia affects functional activities and participation in social roles years after onset. Some group and individual programs have reported success in improving social activities and perceived wellness. These programs typically last several weeks or months. A short, 2-day seminar style program designed for adults with chronic aphasia and their families is described in this report. Six-month follow-up data from participant pairs demonstrates a significant improvement in functional activity level, improved knowledge of aphasia, and improved family relationships. Nonparticipant pairs did not demonstrate any changes over the same period. These findings replicate and extend those of a previous study on the same 2-day seminar [Top. Stroke Rehabil. 2 (1995) 53.]. The results of this study demonstrate the important outcomes of even a very brief program designed to address the long-term psychosocial needs of adults living with aphasia and their families. EDUCATIONAL OBJECTIVES: As a result of this activity, the participant will be able to: (1) describe a brief family education seminar for adults with chronic aphasia; (2) discuss outcomes in family adjustment, community reintegration, and activities of daily living associated with participation in a family education seminar; and (3) evaluate areas of programming and outcomes that should be addressed in future research. PMID- 11409607 TI - Differentiating dialect from disorder: a comparison of two processing tasks and a standardized language test. AB - Previous research has indicated that norm-referenced language assessment protocols are often biased against dialectal speakers. Recently, the use of processing tasks has emerged as one possible means of reducing this bias in language testing. Processing tasks measure a child's ability to process and manipulate language rather than tap previous linguistic knowledge. The present study utilized 40 subjects between the ages of 7;0 and 7;3 in the following equal groupings: White normal language, White language impaired, African American normal language, African American language impaired. The subjects were administered the Test of Language Development-2P (TOLD-2P), the Nonword Repetition Task (NRT), and the Competing Language Processing Task (CLPT). Results indicated that all three measures differentiated normal-language and language impaired subjects from one another. With regard to cultural group, confirmed speakers of African American English (AAE) with normal language scored significantly lower on the TOLD-2P compared to White normal-language subjects. Scores of the AAE-speaking subjects with normal language on the NRT and CLPT, however, did not differ significantly from the White normal-language subjects. These results suggest that AAE speakers with normally developing language (LN) may be at a disadvantage on tests of prior language knowledge and that processing tasks may be a useful tool in combination with other assessment measures to make less biased clinical decisions. EDUCATIONAL OBJECTIVES: As a result of this activity, the reader will (1) be able to determine the utility of processing tasks in culturally unbiased language assessment. (2) The reader will be able to discriminate the difference between the results of a standardized language test and processing tasks on speakers of AAE. PMID- 11409608 TI - The functional anatomy of the female pelvic floor and stress continence control system. AB - This paper provides an overview of the functional anatomy of the structures responsible for controlling urinary continence under stress. The stress continence control system can be divided into two parts: the system responsible for bladder neck support, and the system responsible for sphincteric closure. Age and injury-related changes in each of these systems are discussed. Understanding the pathophysiology of incontinence on the anatomical level will help to lead to identification of specific defects, thereby allowing better individualized treatment for the incontinent patient. PMID- 11409609 TI - Low urethral pressure profile--clinical implications. AB - The clinical impact of urethral pressure profilometry (UPP) has been a main urological topic over the past three decades. Exaggerated expectations and differing techniques with incongruent results caused a controversial appearance. However, since the UPP is the only method to measure directly aspects of the urethral closure function, all types of sphincteric urinary incontinence represent indications for this measurement. Based on long-term experience this paper aims to discuss the various complexes regarding the urethral pressure profilometry. The significance of the urethral closure function for urinary continence and the urodynamic relevance of active and passive pressure transmission are described. In addition, clinical implications in terms of therapeutic strategies to treat urinary stress incontinence with regards to a low urethral pressure profile are presented. PMID- 11409610 TI - Morphological and physiological characteristics of urethral circular and longitudinal smooth muscle. AB - The urethral wall contains circular and longitudinal smooth muscle. Both layers can develop spontaneous tone, and contract further or relax in response to excitatory or inhibitory stimuli. In the pig the cells do not generate action potentials, and the membranes possess L-type calcium channels and a variety of potassium channels which may modulate the membrane potential and tone. The importance of these layers in generating the urethral pressure is not well understood in the human, but it seems likely that the circular smooth muscle is involved in generating urethral pressure in the pig. PMID- 11409611 TI - Role of calcium and calcium modulation in the control of urethral tone. PMID- 11409612 TI - Neurotransmission and drug effects in urethral smooth muscle. AB - Urethral smooth muscle tension is one of the factors contributing to the maintenance of intraurethral pressure and to continence. This tension is controlled by nervous mechanisms involving a complex pattern of efferent and afferent signalling in parasympathetic, sympathetic, and somatic nerves, with resulting release of contraction- and relaxation-producing transmitters and mediators. Sympathetic activity, via release of NA and stimulation of urethral smooth muscle alpha1-ARs, is the main contraction-producing factor. Among the three high affinity alpha1-AR subtypes identified in molecular cloning and functional studies, the alpha1A subtype seems to predominate in both the male and female urethra. The physiological role of the urethral beta-adrenoceptors has not been established, and the importance of muscarinic receptors (and subtypes) for urethral function is presently unclear. Nitric oxide, produced by nitric oxide synthase within cholinergic nerves, seems to be the predominant inhibitory neurotransmitter, but there is good evidence for the existence of other, as yet unidentified, non-adrenergic, non-cholinergic inhibitory messengers. Vasoactive intestinal polypeptide, adenosine 5'-triphosphate, and carbon monoxide (CO) all relax urethral smooth muscle, and are potential candidates for the nonadrenergic, noncholinergic relaxation, but their roles for urethral function remain to be defined. PMID- 11409613 TI - Neural control of the urethra. AB - Coordination between the urinary bladder and the urethra is mediated by multiple reflex pathways organized in the brain and spinal cord. Some reflexes promote urine storage; whereas other reflexes facilitate voiding. During bladder filling, activation of mechanoreceptor afferent nerves in the bladder wall triggers firing in the cholinergic efferent pathways to the external urethral sphincter (EUS) and in sympathetic adrenergic pathways to the urethral smooth muscle. These storage reflexes are dependent upon interneuronal circuitry in the spinal cord. During voiding the spinal storage reflexes are inhibited by supraspinal mechanisms which originate in the pontine micturition center. Glutamatergic, serotonergic and alpha, adrenergic excitatory transmission as well as GABAergic/glycinergic inhibitory transmission have been implicated in the central control of sphincter reflexes. During voiding, a parasympathetic nitrergic inhibitory input to the urethral smooth is activated. This reflex mechanism which is triggered by bladder afferents persists in paraplegic rats and therefore must be mediated at least in part by spinal interneuronal circuitry. In female rats, the parasympathetic nitrergic pathway is prominent; but in male rats it is obscured by a dominant parasympathetic cholinergic excitatory input to the urethral smooth muscle. The function of the cholinergic pathway in voiding is uncertain. Stimulation of urethral afferents can also influence bladder activity. Contraction of the external urethral sphincter activates afferents that inhibit reflex bladder contractions; whereas infusion of fluid through the urethra facilitates bladder contractions. These reflexes are also organized in the spinal cord and presumably play a role in urine storage and elimination. Alterations in primitive bladder-to urethra and urethra-to-bladder reflex mechanisms may contribute to neurogenic bladder dysfunction. PMID- 11409614 TI - Some biomechanical aspects of continence function. PMID- 11409615 TI - Urethral pressure measurement--problems and clinical value. AB - Urethral pressure measurements are in use to assess urethral closure and voiding function. The lack of general agreement on an explicit definition of urethral pressure and standardisation of the methodology for measurement has limited the clinical utility of urethral pressure measurements. PMID- 11409616 TI - The clinical value of ambulatory urethral pressure recording in women. AB - The value of ambulatory urethral pressure recording (AUPR) was evaluated by reviewing both data from 116 patients investigated and Ambulatory Urodynamic Monitoring (AUM) tracings. Typical tracings are shown from conditions where we find that AUPR is of value. (i) Stress incontinence where a patient who complains of stress incontinence does not leak during the stress test, but leaks while jumping on a trampoline. (ii) Mixed incontinence--in only 43% of patients complaining of both urge ands stress incontinence mixed incontinence was documented by AUM. Thirty-six per cent of patients complaining of both urge and stress incontinence were found to be stress incontinent and 21% were leaking due to an unstable detrusor. (iii) Urethral closure pressure of 20 cmH2O or less (low urethral closure pressure; LUCP). LUCP was found in 13% of the patients. Patients with LUCP may leak due to stress, unstable detrusor or unstable urethra. We conclude that ambulatory recording of both urethral and bladder pressures in addition to leakage may contribute to a more complete diagnosis and treatment in these conditions. PMID- 11409617 TI - The clinical value of video-urethrography. PMID- 11409618 TI - The innervation and properties of the urethral striated muscle. AB - The striated muscle forms an outer sleeve around the urethra and occupies about 80% of the wall. In humans more than 60% of the muscle consists of slow, fatigue resistant type I fibres whereas in male greyhounds only about 6% are type I. Most of the remainder (70% of fibres) are type IIa which are classified as fast but can contract for several hours with little decline in amplitude. Stimulation of pudendal but not pelvic nerves produces contraction of the striated muscle and a rapid increase in urethral pressure. Pelvic nerve branches enter the striated muscle sleeve but pass through it to supply the inner layer of smooth muscle. The striated muscles are well suited to maintain contraction necessary for continence. PMID- 11409619 TI - The clinical value of sonographic imaging of the urethrovesical anatomy. AB - Ultrasound is a complementary evaluation for the study of female urinary incontinence which allows functional-morphological documentation. With sonography dynamic images during coughing, Valsalva, pelvic floor contraction and micturition are provided. Ultrasonography of the urethrovesical anatomy improves our thinking of function and anatomy and will therefore have an important place in the diagnosis and treatment of urogynecologic disorders. PMID- 11409620 TI - The clinical value of dynamic magnetic resonance imaging in normal and incontinent women--a preliminary study on micturition. AB - In this preliminary study in nine volunteers and nine women with genuine stress incontinence (GSI) dynamic magnetic resonance imaging (MRI) was used to study the voiding phase in the sitting position after physiological filling of the bladder by urine. The MRI technique has been documented as being useful for this purpose, but in this small group of women it was not possible to determine any specific difference in the voiding pattern between the volunteers and the G PMID- 11409621 TI - Magnetic resonance imaging of the pelvic floor--possibilities and present status. AB - MRI has had both an increasing availability and fuctionality during the last 5-10 years. In diagnosing problems of the pelvic floor, it has been shown to have a potential, but is at present not to be considered a routine examination. Today's use of MRI of the pelvis includes anatomical/topographical images of high quality, but to make it a valuable diagnostic tool, functional imaging is mandatory. Functional MRI with a potential of simultaneously examining defecation, micturition, bladder motion and pelvic floor muscles, seems promising, but with having practical problems. The use of open MRI systems reduce the practical problems, but does not give the optimum image quality. In the future it is likely that MRI will have an increasing importance concerning diagnoses of the pelvic floor. However, clinical research is needed to evaluate the possible diagnostic gains, and maybe even open for more use of MRI. PMID- 11409622 TI - Stress incontinence observed with real time sonography and dynamic fastscan magnetic resonance imaging--insights into pathophysiology. AB - Our concepts of pathophysiology of stress urinary continence have been greatly shaped by developments in radiographic imaging. Simple radiographs with and without contrast initially revealed the importance of urethral descent in pathogenesis. More recently, magnetic resonance imaging (MRI) and real time ultrasonography are showing soft tissue detail within both a global pelvic and a local urethral context. Careful examination of these studies can extend our concepts of pathophysiology and lead us beyond existing paradigms. We propose a unified theory of stress incontinence based on our dynamic fastscan MRI and real time ultrasonograms of stress incontinence, incorporating known details of pelvic anatomy, sphincteric location and function. The hypothesis introduces the concept of a continence threshold at which the urethra is subjected simultaneously to both shearing and explusive forces. If these forces are sufficient to overcome urethral coaptation at threshold, leakage results. The model proposes an anatomical sequence of changes through which the incontinent urethra cycles between periods of rest and increased abdominal pressure, and suggests a way in which repeated episodes of prolpase and urethral traction by shearing forces exerted by the vagina on the urethra may contribute to the development of intrinsic sphincteric deficiency. PMID- 11409623 TI - Stenotherms at sub-zero temperatures: thermal dependence of swimming performance in Antarctic fish. AB - We examined the burst swimming performance of two Antarctic fishes, Trematomus bernacchii and T. centronotus, at five temperatures between -1 degrees C and 10 degrees C. As Antarctic fishes are considered one of the most cold specialised and stenothermal of all ectotherms, we predicted they would possess a narrow thermal performance breadth for burst swimming and a correlative decrease in performance at high temperatures. Burst swimming was assessed by videotaping swimming sequences with a 50-Hz video camera and analysing the sequences frame-by frame to determine maximum velocity, the distance moved throughout the initial 200 ms, and the time taken to reach maximum velocity. In contrast to our prediction, we found both species possessed a wide thermal performance breadth for burst swimming. Although maximum swimming velocity for both T. bernacchii and T. centronotus was significantly highest at 6 degrees C, maximum velocity at all other test temperatures was less than 20% lower. Thus, it appears that specialisation to a highly stable and cold environment is not necessarily associated with a narrow thermal performance breadth for burst swimming in Antarctic fish. We also examined the ability of the Antarctic fish Pagothenia borchgrevinki to acclimate their burst-swimming performance to different temperatures. We exposed P. borchgrevinki to either -1 degrees C or 4 degrees C for 4 weeks and tested their burst-swimming performance at four temperatures between -1 degrees C and 10 degrees C. Burst-swimming performance of Pagothenia borchgrevinki was unaffected by exposure to either -1 degrees C or 4 degrees C for 4 weeks. Maximum swimming velocity of both acclimation groups was thermally independent over the total temperature range of 1 degrees C to 10 degrees C. Therefore, the loss of any capacity to restructure the phenotype and an inability to thermally acclimate swimming performance appears to be associated with inhabiting a highly stable thermal environment. PMID- 11409624 TI - Ionic balance in the freshwater-adapted Chinese crab, Eriocheir sinensis. AB - Ionic regulation by the gills of the freshwater-adapted Chinese crab, Eriocheir sinensis, was examined. The balance of uptake and loss of NaCl in crabs living in freshwater was established. Urine production was measured directly by cannulating the nephropores. Daily urinary loss of Na+ is equivalent to 16% of the haemolymph Na+ content and is substantially higher than that based on data from indirect measurements reported in the literature. Weight and area of anterior and posterior gills are proportional to body weight. The role of the gills in compensating urinary loss by uptake was determined by analysing changes in Na+ and Cl- concentrations in the external medium in which isolated perfused gills were suspended. In posterior gills, salt loss is quantitatively balanced by NaCl net uptake from an external concentration of 1.3 mmol l(-1) NaCl upwards. The transport constant (Kt) for half maximum saturation of net uptake and saturation of NaCl uptake are 1.5 mmol l(-1) and 4 mmol l(-1), respectively. In contrast to previous studies in which tracer fluxes or transepithelial short-circuit currents were determined, our method of direct ion determination shows that no net uptake of Na+ or Cl- occurs in posterior gills in the absence of the respective counter ion, or when uptake of one ion is blocked by a specific inhibitor. Net uptake of Na+ and Cl- was about equal. We conclude that the uptake of the two ions is coupled. The properties of the branchial ion uptake of E. sinensis correlates with the distribution of this crab in river systems. PMID- 11409625 TI - Lymph osmolality and rehydration from NaCl solutions by toads, Bufo marinus. AB - Toads, Bufo marinus, allowed to maintain an ad libitum state of hydration were dehydrated by 10 15% of their standard weight and allowed to rehydrate from either deionized water or from 10 or 50 mmol l(-1) NaCl solutions. Toads rehydrating from the dilute salt solutions recovered a larger fraction of their standard weight than did toads rehydrating from deionized water despite there being a reduced osmotic gradient. Amiloride did not reduce water gain from these solutions. Water uptake from 100 mmol l(-1) sucrose and 50 mmol l(-1) Na gluconate was reduced relative to deionized water by a fraction predicted from the osmotic gradient. Thus, the presence of both Na+ and Cl- are required for the augmentation of water gain from dilute salt solutions. Toads allowed to rehydrate from 120 mmol l(-1) NaCl for 180 min recovered nearly as much water as toads rehydrating from deionized water for 120 min and the lymph osmolality was not reduced relative to the dehydrated condition. The recovery of water from the salt solution was greater than that predicted from the reduced osmotic gradient and amiloride partially inhibited the rehydration from 120 mmol l(-1) NaCl. Solute coupled water transport can therefore be demonstrated in living animals but only from a NaCl solution that is nearly isoosmotic with the lymph. The mechanism for enhanced water gain from dilute salt solutions remains unresolved. PMID- 11409626 TI - Differential catabolism of muscle protein in garden warblers (Sylvia borin): flight and leg muscle act as a protein source during long-distance migration. AB - Samples of flight and leg muscle tissue were taken from migratory garden warblers at three different stages of migration: (1) pre-flight: when birds face an extended flight phase within the next few days, (2) post-flight: when they have just completed an extended flight phase, and (3) recovery: when they are at the end of a stop-over period following an extended flight phase. The changes in body mass are closely related to the changes in flight (P<0.001) and leg muscle mass (P<0.001), suggesting that the skeletal muscles are involved in the protein metabolism associated with migratory flight. From pre- to post-flight, the flight and the leg muscle masses decrease by about 22%, but are restored to about 12% above the pre-flight masses during the recovery period. Biochemical analyses show that following flight a selective reduction occurred in the myofibrillar (contractile) component of the flight muscle (P<0.01). As this selective reduction accounts only for a minor part of the muscle mass changes, sarcoplasmic (non-contractile) and myofibrillar proteins of both the flight and leg muscle act as a protein source during long-distance migration. As a loss of leg muscle mass is additionally observed besides the loss in flight muscle mass, mass change seems not to be strictly associated with the mechanical power output requirements during flight. Whereas the specific content of sarcoplasmic proteins in the flight muscle is nearly twice as high as that in the leg muscle (P<0.001), the specific content of myofibrillar proteins differs only slightly (P < 0.05), being comparably low in both muscles. The ratio of non-contractile to contractile proteins in the flight muscle is one of the highest observed in muscles of a vertebrate. PMID- 11409627 TI - A Na+-dependent electrogenic glutamate transporter current in voltage-clamped cells of corpora allata in the cricket Gryllus bimaculatus. AB - Application of L-glutamate (1 mM) to corpora allata cells of the adult male cricket Gryllus bimaculatus caused a membrane depolarization of 5.9+/-0.3 mV (mean +/- SE) from a resting potential of -62.2+/-1.3 mV (n=57). The underlying mechanism for this depolarization was studied by applying the two-electrode voltage-clamp technique. Application of L-glutamate (1 mM) elicited an inward current that peaked at 8.1+/-0.7 nA (n = 73) at a holding potential of-50 mV. Both L- and D-aspartate also induced an inward current of almost the same amplitude as L-glutamate, whereas D-glutamate failed to induce an inward current. Glutamate receptor agonists, such as kainate, quisqualate, alpha-amino-3-hydroxy 5-methyl isoxazole-4-propionic acid, and N-methyl-D-aspartate, were ineffective in eliciting inward currents. The glutamate-induced inward current did not reverse even when the holding potential was set to +40 mV. The replacement of extracellular Na+ with choline+ eliminated the inward current. These results strongly suggest that the current induced by glutamate is mediated by a glutamate transporter rather than a glutamate receptor. We further examined the effects of 12 amino acid analogs which are known to be selective inhibitors of the mammalian excitatory amino acid transporters (EAATs) on the corpora allata transporter. From the effects of these inhibitors, we conclude that the glutamate transporter expressed in corpora allata cells of the cricket is similar to the high affinity glutamate transporters cloned from human brain, especially EAAT1 and EAAT3. Unlike mammalian transporters, however, serine-O-sulfate has the most potent action, suggesting the unique feature of the glutamate transporter expressed in the corpora allata. PMID- 11409628 TI - Free fatty acids digested from pollen and triolein in the honeybee (Apis mellifera carnica Pollmann) midgut. AB - Honey bees satisfy their lipid requirement by consuming pollen. The free fatty acid content of the midgut was used to quantify fat digestion. Midguts extracted from younger workers of known ages and from foragers were divided into three components: endoperitrophic region (peritrophic membrane with gut contents), extraperitrophic region and intestinal wall. Both the total amount of pollen and the amount of free fatty acids in the endoperitrophic region and in the intestinal wall depend on the bee's age. The amounts increase within the 1st 3 days of a honey bee's life, reach maxima around the age of 8 days and then decrease continuously to the lowest values, measured in forager bees. Forced feeding with triacylglycerol results in significantly higher levels of free fatty acids, especially in the endoperitrophic region, in 8-day-old bees and foragers. This indicates that lipolytic activity depends on age and that the free fatty acid content in 8-day-old bees is primarily limited by the amount and availability of lipids ingested. The results show further that fat digestion depends on the functional status of honey bees, as is the case for pollen consumption, speed of transport of pollen bolus through the alimentary canal and protein digestion. PMID- 11409629 TI - Plasticity of myosin heavy chain expression with temperature acclimation is gradually acquired during ontogeny in the common carp (Cyprinus carpio L.). AB - Common carp (Cyprinus carpio L.) were reared from hatching until 61 mm total length (TL) at 21 degrees C. At 14 weeks and 20 weeks post-hatch, corresponding to initial lengths of 30 mm and 44 mm respectively, fish were acclimated to 10 degrees C using a rate of cooling of 1 degrees C per day. A statistical model was used to compare the time course in the change of white muscle myofibrillar ATPase activity with temperature acclimation. The myosin heavy chain (MHC) composition of white muscle myofibrils was investigated using peptide mapping. A significant increase in myofibrillar ATPase activity was observed after 2-3 weeks in the 44 mm group, but not until 4-5 weeks in the 30 mm group. when they had reached 37 mm TL. The MHC banding pattern of 120 mm TL fish acclimated to 10 degrees C or 21 degrees C for a minimum of 6 weeks were distinct from each other. The MHC peptide map characteristic of 10-degrees C-acclimated fish was not observed in individuals less than 37 mm length. We therefore conclude that the capacity to alter the composition and properties of myofibrils with cold acclimation is acquired in juvenile carp at around 37 mm TL. PMID- 11409630 TI - Correlations of plasma lipid metabolites with hibernation and lactation in wild black bears Ursus americanus. AB - During the denning period, black bears (Ursus americanus) are capable of enduring several months without food. At the same time, female bears that are pregnant or lactating have an added metabolic stress. Based on laboratory studies, much of the energy required to support metabolism and lactation during denning in black bears comes from lipid reserves. These lipid reserves are mobilized and the most metabolically active lipid fraction in the blood are nonesterified fatty acids (NEFA). Therefore, we hypothesized that plasma NEFAs would be higher in denning relative to active bears and in lactating relative to non-lactating female bears. We further hypothesized that in bears with elevated plasma NEFA levels, other lipid-related parameters (e.g., ketone bodies, albumin, cholesterol, lipase) would also be elevated in the plasma. Denning bears had significantly increased NEFA levels in all classes (saturates, monoenes, and polyenes). A doubling of plasma NEFA levels and a 33% increase in albumin, the plasma fatty acid binding protein, in denning bears, resulted in NEFA/albumin ratios that were higher in denning bears (4:1) compared to those of active bears (3:1). Bears became relatively ketonemic with a 17-fold increase in D-beta-hydroxybutyrate levels during the denning period. Plasma cholesterol approximately doubled and lipase was ten-fold lower in denning relative to active bears. These findings indicate a strong correlation between plasma lipid metabolites and the denning period in a wild population of black bears. PMID- 11409631 TI - Physiological responses to supercooling and hypoxia in the hatchling painted turtle, Chrysemys picta. AB - We investigated physiological responses to supercooling in hatchling painted turtles (Chrysemys picta) which remain in their natal nests over winter and therefore may become exposed to subzero temperatures. These turtles are freeze tolerant but also must rely on supercooling to survive exposure to the lower temperatures occurring in nests during winter. We compared whole-body concentrations of lactate, glucose, glycerol, and ATP in turtles chilled at 0 degrees C, -4 degrees C, or -6 degrees C for 5 days, or at 6 degrees C for 19 days. In a companion experiment, we measured metabolite concentrations in turtles exposed to a hypoxic environment for 1 day, 4 days, or 8 days. Supercooling and hypoxia exposure were both associated with an increase in concentrations of lactate and glucose and a decrease in glycerol concentrations (albeit no change in the ATP pool), suggesting that supercooling induces functional hypoxia. We conclude that hypoxia tolerance may be an important pre-adaptation for surviving exposure to subzero temperatures in hatchling C. picta. PMID- 11409632 TI - The calibre of the foramen of Panizza in Crocodylus porosus is variable and under adrenergic control. AB - The foramen of Panizza is located within the outflow tract of the crocodilian heart, between the left and right aortas. It has been suggested that the foramen of Panizza has a variable calibre, which could explain the profound changes in the distribution of flows and pressure profiles recorded in the right and left aortas. We investigated this possibility using a modified in-situ perfused heart preparation in combination with isolated strip preparations from the outflow tract. In the perfused heart preparation, bolus injections of adrenaline increased the resistance in the foramen of Panizza, indicating a decrease in its diameter. Isolated strip preparations from the outflow tract showed a concentration-dependent increase in tension in response to adrenaline, while vasoactive intestinal polypeptide caused a relaxation in adrenaline pre contracted strip preparations. We propose that an increase in the diameter of the foramen of Panizza may be important during pulmonary to systemic shunts to allow blood to flow from the left to right aorta (reverse foramen flow) in order to supply the carotid and coronary arteries. During non-shunting conditions, a constricted foramen may prevent excess flow from the right to left aorta during diastole. PMID- 11409633 TI - Cytogenetic study of Callicebus hoffmannsii (Cebidae, Primates) and comparison with C. m. moloch. AB - Callicebus is a neotropical primate genus divided into four or five groups of species. Species of the moloch group are distributed in the tropical forests of the Amazon basin. The karyotype of Callicebus hoffmannsii (moloch group) was studied by means of G- and C-banding, Ag-NOR staining and in situ hybridization of telomeric probes. C. hoffmannsii had 2n = 50 chromosomes, with ten biarmed and fourteen acrocentric autosomal pairs. The X chromosome was submetacentric and the Y chromosome was a minor acrocentric. Constitutive heterochromatin was detected in the centromeric regions of all chromosomes; in pairs 7 and 10, it was found in the distal regions of the short arms, and distally in the long arm of the X chromosome. Size heteromorphism in C-bands was detected in pairs 7 and 10. Ag-NOR staining revealed a maximum of three nucleolar organizers. Telomeric probes hybridized only at the terminal regions of all chromosomes. Additionally, a comparison was carried out between C. hoffmannsii and C. m. moloch (2n = 48), as previously reported. Both species shared gross chromosomal similarities diverging by a single rearrangement of centric fusion/fission. A high similarity between C. hoffmannsii and C. donacophilus indicated a close association between the moloch and donacophilus groups. PMID- 11409634 TI - Influence of hydrocortisone and microwave radiation on the mechanical characteristics of rat bone tissue. AB - This work deals with the mutual action of hydrocortisone and low intensity microwave radiation (MWR) on the bone tissue of rats. The bone density and velocity of ultrasound was measured in order to evaluate the Young's modulus of the femur. The results show a stimulating effect of the low-intensity MWR field on regeneration of the bone tissue of rats. The MWR, during a long application of hydrocortisone, may be a characteristic protective factor for the bone tissue. PMID- 11409635 TI - Microscopical aspects of accessory air breathing through a modified stomach in the armoured catfish Liposarcus anisitsi (Siluriformes, Loricariidae). AB - The presence of an accessory air breathing mechanism as verified by several authors, is widespread among Loricariidae, where modified parts of the digestive tract act primarily as oxygen-exchange organs. An anatomical and histological analysis was carried out on the stomach and intestine of the armoured catfish Liposarcus anisitsi. The data support the assumption that the modified stomach is responsible for holding air and allows blood oxygenation under hypoxia. Experiments demonstrating survival of air breathing Liposarcus in severely hypoxic water support the hypothesis and are discussed. PMID- 11409636 TI - Analysis of the distribution pattern of sister chromatid exchanges in mitotic chromosomes of human T-lymphocytes. AB - Band locations of 344 sister chromatid exchanges (SCE), observed in chromosome preparations of peripheral T-lymphocyte cultures, were identified using a sequential Q-banding-sister chromatid differentiation staining protocol. A comparison between expected Poisson, expected negative binomial distributions and observed SCE patterns, revealed that the negative binomial distribution provided a better fit to our SCE data. The aggregated pattern of SCE distribution indicated that some bands were more prone to undergo these events. Genetic markers assigned to those bands recording a significant number of exchanges seemed to correspond with the transcriptionally active genes. The results confirm that SCE are induced at the early S-phase of the cell cycle involving the early replicating genes in these events. Potential applications of cytogenetic techniques employed in the present study are proposed. PMID- 11409637 TI - Cytophotometric estimation of in situ DNA content in several species of Araceae. AB - A wide variation in the in situ 4C DNA content, ranging from 15.02 pg to 54.09 pg was found in thirteen genera of the family Araceae. The obligate perennial species showed greater 4C DNA values compared with the facultative perennials and annuals. A remarkable heterogeneity in 4C nuclear DNA amounts was noted among obligate perennials. Intraspecific constancy in the amount of 4C DNA was recorded. The nuclear DNA content correlated positively with chromosome size, duration of the mitotic cycle, and annual to perennial growth forms. Despite an extensive variation in DNA content among the aroids investigated, each species was distinctly characterized by its specific nuclear DNA value, indicating its usefulness in taxonomic characterization and comparison of different aroids. PMID- 11409638 TI - Antihyperlipidaemic and antihypercholesterolaemic effects of Anethum graveolens leaves after the removal of furocoumarins. AB - Serum triacylglycerides and total cholesterol levels in rats, with hyperlipidaemia induced by diet, were determined after oral adminstration of a water extract of Anethum graveolens leaves before and after the extraction of the furocoumarin content of the leaves. Administration of the extracts consecutively for 14 days reduced the triacylglycerides and total cholesterol levels by almost 50 and 20%, respectively. Chloroform extraction of furocoumarins from the aqueous extracts did not reduce the antihyperlipidaemic potential of the extracts to a significant degree. Oral administration of the essential oil of A. graveolens seeds, at two different doses, also reduced the triacylglyceride levels by almost 42%. The total cholesterol level was not reduced by the same doses of the essential oil. PMID- 11409639 TI - Local angiotensin II-generating system in vascular tissues: the roles of chymase. AB - Roles of each angiotensin II producing enzymes of each of the angiotensin II producing enzymes were reviewed based on experimental models. In vascular tissues, angiotensin II is potentially cleaved from angiotensin I by angiotensin converting enzyme (ACE) and chymase. It has been confirmed that vascular tissues of humans, monkeys, dogs and hamsters have a chymase-dependent angiotensin II forming pathway. Much like other hypertensive models, hamster hypertensive models show high levels of vascular ACE activity, but not chymase activity. In hypertensive hamsters, administration of either an ACE inhibitor or an angiotensin II type 1 (AT1) receptor antagonist resulted in similar reductions in blood pressure, suggesting that chymase is not involved in the maintenance of high blood pressure in this model. In monkeys fed a high-cholesterol diet, ACE activity was increased in the atherosclerotic lesions, and an ACE inhibitor and an AT1 receptor antagonist prevented atherosclerosis to a similar degree, suggesting that ACE may be mainly involved in the development of atherosclerosis. After balloon injury in dog vessels, both ACE and chymase activities were locally increased about 3-fold in the injured arteries, and an AT1 receptor antagonist was effective in preventing the intimal formation, but an ACE inhibitor was ineffective. In dog grafted veins, the activities of chymase were increased 15 fold, but those of ACE were increased only 2-fold, and the intimal formation was suppressed by either an AT1 receptor antagonist or a chymase inhibitor. In the normal vascular tissues, ACE plays a crucial role for angiotensin II production, whereas chymase is stored in mast cells in an inactive form. Chymase acquires the ability to form angiotensin II following mast cells activation followed by mast cells activation by a strong stimulus such as occurs in catheter-injury or grafting. Together, these results indicate that chymase plays a major role in the vascular angiotensin II-generating system, particularly in cases of vascular injury. PMID- 11409640 TI - Diurnal variation of hemodynamic indices in non-dipper hypertensive patients. AB - The purpose of this study was to elucidate the underlying mechanisms of blunted nocturnal blood pressure reduction in non-dipper hypertensive patients. We studied the diurnal variations in systemic hemodynamic indices and baroreflex sensitivity. In 45 subjects with essential hypertension (24 men; mean age, 49+/-1 years), intra-arterial pressure was monitored telemetrically. Non-dippers were defined as those with a nocturnal reduction of systolic blood pressure of less than 10% of daytime systolic blood pressure. Stroke volume was determined using Wesseling's pulse contour method, calibrated with indocyanine green dilution. Baroreflex sensitivity was calculated as deltapulse interval/deltasystolic blood pressure on spontaneous variations. The mean values of the hemodynamic parameters were calculated every 30 min. Twenty-six subjects were classified as non-dippers. Daytime blood pressure was not significantly different between dippers (149+/ 4/87+/-3 mmHg) and non-dippers (147+/-3/82+/-2 mmHg), while the nighttime blood pressure was significantly reduced in dippers (131+/-3/77+/-2 mmHg) but not in non-dippers (145+/-3/80+/-2 mmHg). Nocturnal decreases in both cardiac index and stroke index were smaller in non-dippers (-12.0+/-1.2% and 1.5+/-1.0%) than in dippers (-17.5+/-1.4% and -2.2+/-1.1%). Baroreflex sensitivity significantly increased at nighttime both in dippers (6.5+/-0.6 to 8.0+/-0.7 ms/mmHg) and in non-dippers (5.1+/-0.3 to 6.4+/-0.4 ms/mmHg). Neither daytime nor nighttime baroreflex sensitivity was significantly different between the groups. We conclude that the hemodynamics of non-dipper essential hypertension are characterized by an inadequate nocturnal decrease in cardiac index and stroke index, suggestive of relative volume expansion or malsuppressed sympathetic activity. PMID- 11409641 TI - Profiles of patients who control the doses of their antihypertensive drugs by self-monitoring of home blood pressure. AB - The present study profiled patients who control doses of antihypertensive drugs by themselves based on self-monitoring of their blood pressure (self controllers). A total of 1,028 consecutive outpatients who were taking antihypertensive drugs and who were attending the cardiovascular outpatient clinic of our institute responded to a questionnaire in 1998. They were asked how often they measured their blood pressure, how often they missed taking their medication, and whether or not they had a chance to adjust the doses of antihypertensive drugs by themselves based on self-monitored blood pressure. The frequency of self-controlling of antihypertensive drugs was also examined in 918 patients on antihypertensive drugs in 1997. In 1997, 23 of 918 patients (2.5%) were self-controllers, and 26 of 1,028 patients (2.5%) were self-controllers in 1998. The frequency of home blood-pressure measurement was significantly greater in self-controllers than in the remaining patients (non self-controllers) (p<0.01). The prevalence of proteinuria was significantly less in the former than in the latter. Prior to the start of antihypertensive drugs, blood pressure was significantly lower for the self-controllers (154.4+/-3.8/96.4+/-1.4 mmHg) than for the non self-controllers (169.3+/-0.7/101.7+/-0.4 mmHg) (p<0.001). Clinically measured blood pressures did not differ significantly between the self controllers and non self-controllers. Thus, about 2.5% of patients on antihypertensive drugs controlled their drug doses by themselves based on self monitoring of their blood pressure. These patients were characterized by having a milder form of hypertension and by more frequent home blood-pressure measurement than non self-controllers. PMID- 11409642 TI - Cardiovascular and sympathetic responses to dental surgery with local anesthesia. AB - We investigated changes in blood pressure and blood variables, including plasma catecholamines, serum glucose and insulin concentrations, during dental surgery. The study included 11 normotensive patients (age, 22.5+/-0.7 years) who underwent tooth extraction at Kyushu Dental College Hospital. Three to 7 days prior to dental surgery, blood pressure, pulse rate, and heart rate variability were measured every 30 min over 24 h. The low frequency (LF: 0.05 to 0.15 Hz) and high frequency (HF: 0.15 to 0.40 Hz) powers were calculated, and the ratio of LF to HF (LF/HF) and HF were used as indexes of sympathetic and parasympathetic activities, respectively. Lidocaine, 2% with epinephrine (1:80,000), was used as the local anesthetic for all patients. Systolic blood pressures significantly increased during dental surgery (+10.8+/-3.5 mmHg); however, this increase failed to correlate not only with baseline systolic blood pressure but with 24-h averaged blood pressures, LF/HF or HF. On the other hand, plasma epinephrine and norepinephrine concentrations increased during dental surgery, and peak values of these variables were obtained after local anesthesia and during tooth extraction, respectively. Serum glucose level increased after local anesthesia (control vs. local anesthesia: 5.16+/-0.11 vs. 5.62+/-0.10 mmol/l; p<0.01); however, plasma insulin concentrations did not change significantly. These results suggest that 1) ambulatory measurements of blood pressure and heart rate variability over 24 h cannot predict the responses of blood pressure during dental surgery, and that 2) administration of local anesthetic and tooth extraction activate sympathoadrenal outflow, resulting in an increase in serum glucose level in normotensive subjects. PMID- 11409643 TI - Non-Linear trends in the blood pressure of Japanese adults. AB - Hypertension is one of the risk factors for the development of coronary artery disease as well as stroke. The National Nutrition Survey shows that the systolic and diastolic blood pressures of Japanese adults have decreased over time. These trends have plateaued during the last decade. We investigated national trends in blood pressure (BP) levels during these 10 years by plotting the relation between the mean BP levels during year t, BP(t), and those of the following year, BP(t+1). When we plotted the systolic BP levels of Japanese men and women aged 15 to 80 years, the trace revolved clockwise, with cycles of 4 to 5 years. The approximate center of the circular trace corresponding to data for 1985 through 1990 was at 134 mmHg in men and 129 mmHg in women, and the approximate center of a smaller circle was at 135 mmHg in men and 130 mmHg in women, respectively. In addition, with respect to the data for Japanese men in their 60s, the mean systolic BP level decreased linearly from 1982 to 1986, whereas it seemed to generate 4 circles after 1986. The approximate center of the first circle was 146 mmHg in the men. The center of the second circle was 145 mmHg and that of the third circle was 144 mmHg. Plotting of the data for Japanese women of the same age group yielded the same number of circles as the data for men. Our method of documenting changes in the mean BP levels in Japanese adults provided information about a chaotic oscillation that occurred in the populations we studied. The centers of Lorenz-plotted circles may thus provide essential information about trends in the BP levels in these populations. PMID- 11409644 TI - Structure and function of the left ventricle and carotid artery in hemodialysis patients. AB - Hemodialysis patients frequently show associated hypertension, which can lead to a number of cardiovascular complications. The aim of this study was to assess the effects of hypertension on the structure and function of the carotid artery and left ventricle (LV) in hemodialysis patients. In addition, we investigated the contribution of hemodialysis and other risk factors. Fifty-two hemodialysis patients, 71 hypertensive patients (HT group) and 30 normotensive subjects (NT group) were included in this study. Hemodialysis patients were divided into two groups: 35 patients with hypertension (HDHT group), and 17 patients without hypertension (HDNT group). We measured intima-media thickness (IMT), plaque score, end-diastolic diameter, and stiffness index beta of the carotid artery by ultrasonography, and LV mass index (LVMi), endocardial fractional shortening (FS), and midwall FS (MWS) by echocardiography. A multiple stepwise regression analysis including hemodialysis, hypertension, diabetes mellitus, and other risk factors was also performed. IMT was significantly higher in the HT and HDHT groups than in the NT group. Plaque score and diameter of the carotid artery were higher in the HDHT group than in the other three groups. The stiffness index beta was higher in the HDHT group than in the non-hemodialysis groups. In multivariate analysis, IMT was independently correlated with age and hypertension. Plaque score and stiffness index beta were independently associated with age, hypertension, and hemodialysis. LVMi was higher in HT and hemodialysis-patients groups than in the NT group. Hypertension and hemodialysis were strong and independent predictors of LVMi. FS showed no significant differences among the four groups, but MWS was significantly lower among the hemodialysis patients than in the NT group. MWS was independently correlated with hemodialysis and diabetes. In conclusion, hemodialysis per se advanced both atherosclerosis and arteriosclerosis of the carotid artery. Moreover, it increased LVMi and caused cardiac dysfunction. Associated hypertension might thus accelerate the progression of atherosclerosis and arteriosclerosis of the carotid artery and the increase of LVMi. PMID- 11409645 TI - Differential effects among thiazolidinediones on the transcription of thromboxane receptor and angiotensin II type 1 receptor genes. AB - Peroxisome proliferator-activated receptor (PPAR)-gamma ligands thiazolidinediones (TZDs) have recently been reported to be anti-hypertensive and anti-atherosclerotic. We have previously shown that one of the TZDs troglitazone significantly suppressed the transcription of both thromboxane receptor (TXR) and angiotensin II type 1 receptor (AT1R) genes in vascular smooth muscle cells (VSMCs) by activating PPAR-gamma. In the present study, we compared the effects of troglitazone and other TZDs on the transcription of these genes. TXR and AT1R mRNAs in rat VSMCs were determined by semi-quantitative RT-PCR. Luciferase chimeric constructs containing either the 989-bp rat TXR gene promoter or the 1,969-bp rat AT1R gene promoter were transiently transfected into VSMCs. The cells were incubated with troglitazone, RS-1455 (a derivative of troglitazone which does not contain the hindered phenol resembling alpha-tocopherol), pioglitazone, or rosiglitazone for 12 h before harvesting. mRNA expression levels of TXR and AT1R were significantly decreased by troglitazone in contrast to rosiglitazone. TXR gene and AT1R gene transcription was significantly suppressed by troglitazone in a dose-dependent manner, while RS-1455 was less potent. Pioglitazone and rosiglitazone weakly suppressed the transcription of both genes in a manner almost similar to RS-1455. We have shown that troglitazone suppresses transcription of both the TXR and AT1R genes more potently than other TZDs. The structure of troglitazone and RS-1455 is identical except the hindered phenol, which is recently recognized to function as an antioxidant. Moreover, we have shown that the potency for activating PPAR-gamma is almost identical between troglitazone and RS-1455. We therefore speculate that the strong transcriptional suppression of the TXR and AT1R genes by troglitazone may be mediated in part by its antioxidant effect. PMID- 11409646 TI - Trandolaprilat, an angiotensin-converting enzyme inhibitor, is not excreted in bile via an ATP-dependent active transporter (cMOAT). AB - Trandolapril is the prodrug of an angiotensin-converting enzyme (ACE) inhibitor. It has been proposed that its active metabolite, trandolaprilat, is mainly excreted in bile, but this has not been clearly demonstrated. Recently it has been reported that temocaprilat, an active metabolite of the ACE inhibitor temocapril, is effectively excreted in bile via an ATP-dependent active transporter (canalicular multispecific organic anion transporter: cMOAT). To investigate whether trandolaprilat has the pharmacological ability to affect the cMOAT system in a manner similar to temocaprilat. The lipophilicity of trandolaprilat and temocaprilat was measured to determine the n-octanol-water partition coefficients. The dose-dependent inhibition of the up-take of [3H] estradiol-17beta-D-glucuronide and [3H]-2,4-dinitrophenyl-S-glutathione, which are good substrates for cMOAT, in canalicular membrane vesicles (CMVs) prepared from Sprague-Dawley rats was determined in the presence of trandolaprilat and temocaprilat. The partition coefficient of trandolaprilat (log Po/w - 1.1) was about 30 times higher than that of temocaprilat (log Po/w - 2.5). The uptake of [3H]-estradiol-17beta-D-glucuronide and [3H]-2,4-dinitrophenyl-S-glutathione was dose-dependently inhibited by the presence of temocaprilat, but trandolaprilat had no effect on the transport of [3H]-estradiol-17beta-D-glucuronide or [3H]-2,4 dinitrophenyl-S-glutathione into CMVs even at concentrations as high as 200 microM. It could be concluded that trandolaprilat has a higher lipophilicity than temocaprilat. But the hepatobiliary excretion system via cMOAT may not contribute to the excretion of trandolaprilat in bile. PMID- 11409647 TI - Benidipine inhibits expression of ET-1 and TGF-beta1 in Dahl salt-sensitive hypertensive rats. AB - Endothelin and growth factors such as transforming growth factor (TGF)-beta1 are important regulators of the cardiovascular system. Although increased production of endothelin-1 (ET-1) and TGF-beta1 have been reported in left ventricular hypertrophy, the detailed roles of these substances in hypertrophy remain to be determined. To elucidate the cardioprotective effects of calcium antagonists in left ventricular hypertrophy, we evaluated the effects of long-term treatment with benidipine, a long-acting calcium antagonist, on preproET-1, ET(A) receptor (ETAR) and TGF-beta1 expression in the left ventricle and evaluated the relations between these effects and myocardial remodeling in Dahl salt-sensitive hypertensive (DS) rats fed a high-salt diet. After 5 weeks of feeding an 8% NaCl diet to 6-week-old DS rats (i.e., at 11 weeks of age), a distinct stage of concentric left ventricular hypertrophy (DSLVH) was noted. Benidipine (DSLVH-B group, n= 8; 1 mg/kg/day, subdepressor dose) or vehicle (DSLVH-V group, n=8) was administered to 6-week-old DS rats for 5 weeks, or until the onset of DSLVH stage, and age-matched (11-week-old) Dahl salt-resistant rats fed the same diet served as a control group (DR-C, n=8). Blood pressure was similar between the DSLVH-B and DSLVH-V groups, but was significantly lower in DR-C rats. The preproET-1, ETAR and TGF-beta1 expressions in the left ventricle were significantly higher in DSLVH-V than in DR-C rats, and significantly lower in DSLVH-B than in DSLVH-V. Benidipine administration resulted in significant improvements in the wall-to-lumen ratio and perivascular fibrosis in the coronary arterioles, and in myocardial fibrosis. We therefore concluded that myocardial remodeling and left ventricular hypertrophy in DS hypertensive rats fed a high salt diet were significantly ameliorated by a subdepressor dose of benidipine, and that this amelioration was partly due to decreases in the expression of ET-1 and TGF-beta1 in the left ventricle. PMID- 11409648 TI - Antioxidants inhibit JNK and p38 MAPK activation but not ERK 1/2 activation by angiotensin II in rat aortic smooth muscle cells. AB - Angiotensin II (Ang II) induces vascular smooth muscle cell (VSMC) hypertrophy, which results in several cardiovascular diseases. Ang II-induced cellular events have been mediated, in part, by reactive oxygen species (ROS) which also involve activation of mitogen-activated protein (MAP) kinases. Although it has been proposed that the therapeutic administration of antioxidants is useful for vascular diseases, the precise mechanisms which regulate ROS-sensitive signaling events have not been well characterized. Thus, we hypothesized that antioxidants may affect ROS-mediated MAP kinases activation induced by Ang II. The present findings showed that Ang II stimulated rapid and significant activation of ERK 1/2, JNK and p38 MAPK in cultured rat aortic smooth muscle cells (RASMC). Ang II induced ERK 1/2 activation was not affected by all antioxidants examined, whereas JNK was sensitive to all antioxidants. In contrast, p38 MAPK activation was inhibited by DPI and ascorbic acid concentration-dependently, but by NAC only at high concentration. DETC and Trolox C had no effects on p38 MAPK activation by Ang II. We further examined the effects of antioxidants on Ang II-induced increases in oxygen consumption as an index of ROS generation in RASMC. DPI strongly inhibited Ang II-induced increases in oxygen consumption. DETC also inhibited Ang II-induced oxygen consumption, whereas ascorbic acid markedly augmented it. These findings suggest that the inhibitory effects of antioxidants on MAP kinases activation in VSMC are attributable, in part, to their modulating effects on ROS generation by Ang II in VSMC. Thus, inhibition of MAP kinases by antioxidants may imply their usefulness for relief of cardiovascular diseases. PMID- 11409649 TI - Estriol improves membrane fluidity of erythrocytes by the nitric oxide-dependent mechanism: an electron paramagnetic resonance study. AB - The present in vitro study was performed to investigate the effects of estriol (E3) on membrane fluidity of erythrocytes by means of an electron paramagnetic resonance (EPR) and spin-labeling method. E3 was shown to significantly decrease the order parameter (S) for 5-nitroxide stearate (5-NS) and the peak height ratio (ho/h-1) for 16-NS obtained from EPR spectra of erythrocyte membranes. This finding indicated that E3 might increase the membrane fluidity of erythrocytes. The effect of E3 was significantly potentiated by the nitric oxide (NO) donor, S nitroso-N-acetylpenicillamine (SNAP), and a cyclic guanosine 3',5'-monophosphate (cGMP) analog, 8-bromo-cGMP. In contrast, the change in the membrane fluidity induced by E3 was antagonized by the NO synthase inhibitor, L-NG-nitroarginine methyl-ester (L-NAME), and asymmetric dimethyl-L-arginine (ADMA). The results of the present study showed that E3 significantly increased the membrane fluidity and improved the microviscosity of erythrocyte membranes, partially mediated by an NO- and cGMP-dependent pathway. Furthermore, the data might be consistent with the hypothesis that E3 could have a beneficial effect on the rheological behavior of erythrocytes and may play a crucial role in the regulation of microcirculation. PMID- 11409650 TI - Calcium buffering of resting, voltage-dependent Ca2+ influx by sarcoplasmic reticulum in femoral arteries from spontaneously hypertensive rats at prehypertensive stage. AB - We examined the Ca2+-buffering function of the sarcoplasmic reticulum (SR) in the resting state of arteries from spontaneously hypertensive rats (SHR) at a prehypertensive stage. Differences in the effects of cyclopiazonic acid (CPA) and thapsigargin, agents that inhibit SR Ca2+-ATPase, and of ryanodine, which depletes SR Ca2+, on tension and cellular Ca2+ level were assessed in endothelium denuded strips of femoral arteries from 4-week-old SHR and normotensive Wistar Kyoto rats (WKY). Addition of CPA, thapsigargin or ryanodine to the resting state of the strips caused an elevation of cytosolic Ca2+ level and a contraction in both WKY and SHR. These responses were larger in SHR than in WKY. The contractions were inhibited strongly by 100 nM nifedipine or 3 microM verapamil and were abolished by Ca2+-free solution. Nifedipine, verapamil or Ca2+-free solution itself caused a relaxation from the resting state of SHR strips, but not from that of WKY strips. The resting Ca2+ influx in arteries measured by a 5-min incubation with 45Ca was significantly larger in SHR than in WKY. This influx was decreased by 10 microM CPA or 10 microM ryanodine in both WKY and SHR. These results suggest that in the resting state of the femoral artery from 4-week-old SHR, the greater part of the increased Ca2+ influx via L-type Ca2+ channels is buffered by Ca2+ uptake into the SR, while some Ca2+ reaches the myofilaments, resulting in the maintenance of resting tone. PMID- 11409651 TI - Intravascular ultrasound detects coarctation of the renal artery in a patient with Moyamoya disease. AB - A 19-year-old man with moyamoya disease was diagnosed as having renovascular hypertension, based on stenosis of the proximal portion of the right renal artery with elevated plasma renin activity. Intravascular ultrasound (IVUS) imaging at the renal artery lesion revealed focal narrowing of the renal artery without vascular wall thickening (i.e., coarctation). The coarctation of the renal artery was adequately dilated by stent implantation after suboptimal balloon angioplasty. After the procedure, the patient's hypertension improved. The findings of the present case suggest that IVUS-guided renal angioplasty is an effective therapeutic procedure for correcting coarctation of the renal artery in patients with moyamoya disease. PMID- 11409652 TI - Association study between the variants of the human ANP gene and essential hypertension. AB - Variants of atrial natriuretic peptide (ANP) are reported to be more common in blacks with hypertension than in normotensive controls and constitute an independent risk factor for cerebral infarction. The purpose of the present study was to investigate the role of ANP in the pathogenesis of essential hypertension (EH) in the Japanese. We investigated 2 previously reported ANP gene markers, G1837A and T2238C, for their possible associations with EH. A total of 233 individuals with EH and 213 age-matched normotensive (NT) control subjects were studied. The frequencies of the G and A alleles were 0.09 (42/466) and 0.91 (424/466), respectively, for the NT group and 0.11 (47/426) and 0.89 (379/426), respectively, for the EH group. These frequencies did not differ significantly between the two groups. The frequencies of the T and C alleles were 0.024 (11/466) and 0.97 (455/466), respectively, for the NT group and 0.03 (13/426) and 0.97 (413/426), respectively, for the EH group. These frequencies also did not differ significantly between the two groups. Neither G1837A nor the T2238C polymorphism of the ANP gene was associated with EH. Our findings do not support the hypothesis that the G1837A and T2238C polymorphisms of the ANP gene are markers for EH in the Japanese. PMID- 11409653 TI - Lack of correlation between Mbo I restriction fragment length polymorphism of renin gene and essential hypertension in Japanese. AB - Predisposition to essential hypertension is associated with gene polymorphisms of the renin angiotensin system (RAS). Gene polymorphisms of the angiotensinogen and angiotensin converting enzyme genes are known to be risk factors for hypertension, while few studies concerning the renin gene polymorphism have been published. In the present investigation, we carried out a case control study using a Japanese population to examine the genetic influence of the renin gene on the predisposition to hypertension. Patients (n=235) recruited from outpatients at Osaka University Hospital and diagnosed with essential hypertension or receiving long-term antihypertensive medication participated in the study. Normotensive control subjects (n=510) without a history of hypertension and without diabetes mellitus were recruited from the same population, and were sex matched with experimental subjects. A polymorphism in intron 9 of the human renin gene was determined as the Mbo I restriction fragment length polymorphism (Mbo I RFLP). There was no significant association between Mbo I-RFLP of the renin gene and predisposition to essential hypertension in Japanese (p>0.05, chi2=2.1). These results suggest that the Mbo I (+) allele of the renin gene does not increase the risk for hypertension in Japanese. PMID- 11409654 TI - Genetic variation in the promoter region of the beta2 bradykinin receptor gene is associated with essential hypertension in a Chinese Han population. AB - The present study examined whether a genetic variant (-58T/C) in the promoter region of the human beta2 bradykinin receptor gene was genetically involved in essential hypertension. Chinese hypertensive subjects (n = 120) and normotensive controls (n = 98; sex- and age-matched with hypertensives) were recruited from the outpatients of Fu Wai hospital. Distribution of the -58T/C polymorphism was determined in patients and controls by means of PCR, SSCP, cloning and sequencing. The allelic frequencies were 0.56 for the C allele and 0.44 for the T allele in hypertensive subjects, and 0.46 for the C allele and 0.54 for the T allele in normotensive subjects. The allelic frequencies were in Hardy-Weinberg equilibrium. Significant differences between hypertensive and normotensive subjects were seen in the genotypes distribution (p = 0.045) and allelic frequencies (p = 0.033). These results suggested that -58C allele of the human beta2 bradykinin receptor gene may be an independent risk factor for essential hypertension in the Chinese Han population. PMID- 11409655 TI - Effects of the Chinese medicine Jiang-Tang-Ke-Li on insulin resistance in fructose-fed rats. AB - The aim of this study was to determine the effect of Jiang-Tang-Ke-Li (JTKL), a Chinese medicine used to treat diabetes mellitus, on insulin resistance and hypertension in fructose-fed rats (FFR). Six-week-old male Sprague-Dawley rats were fed either normal rat chow (control) or a fructose-rich chow (FFR) for 6 weeks. For the last 2 weeks of the 6-week period of either diet, the rats were treated by gavage with gum arabic solution as a vehicle (control or FFR) or JTKL (3.24 g/kg/day; FFR+JT), and then an euglycemic hyperinsulinemic glucose clamp technique was performed to estimate insulin sensitivity. Systolic blood pressure was measured each week of the 6-week period. At the end of the glucose clamp, the soleus and extensor digitorum longus (EDL) muscles were dissected out for determination of the role of tumor necrosis factor (TNF)-alpha by an ELISA assay. Systolic blood pressures in the FFR groups were significantly higher than that in the control group, although there was no effect on systolic blood pressure for the last 2 weeks of treatment with JTKL. The average rate of glucose infusion during the glucose clamp, as an index of insulin sensitivity (M value), was significantly lower in the FFR than in the control rats, and treatment with JTKL for 2 weeks significantly increased the M value to that of control. TNF-alpha levels were significantly higher in the soleus and EDL muscles of the FFR (480+/ 46 and 570+/-45 pg/g wet tissue in the soleus and EDL muscles, respectively) than in those of the control rats (177+/-34 and 206+/-33 pg/g wet tissue in the soleus and EDL muscles, respectively; p<0.01). Treatment with JTKL for 2 weeks significantly lowered TNF-alpha levels to the control levels (189+/-22 and 239+/ 92 pg/g wet tissue in the soleus and EDL muscles, respectively). The results suggest that the Chinese medicine JTKL improves insulin resistance and modulates TNF-alpha in the soleus and EDL muscles in hypertensive and insulin-resistant fructose-fed rats. PMID- 11409656 TI - Effect of long-acting isosorbide-5-mononitrate administration on large artery distensibility in patients with essential hypertension. AB - To evaluate the clinical efficacy of long-acting nitrates, isosorbide-5 mononitrate (IS-5-MN), on large artery distensibility in patients with essential hypertension. Large arterial distensibility was assessed by automatic noninvasive measurement of the carotid-femoral pulse wave velocity (PWV). Seventeen patients aged 62.53+/-7.94 years (mean+/-SD) with essential hypertension undering long term antihypertensive therapy were studied in this trial. PWV was measured 2 weeks and 4 weeks after oral administration of IS-5-MN (30 mg once daily) with previous therapy. There was no significant difference in systolic blood pressure, diastolic blood pressure, pulse pressure or heart rate at 2 weeks and 4 weeks after treatment compared with baseline. The carotid-femoral PWV decreased significantly at 2 and 4 weeks after treatment (p<0.05, p< 0.05, respectively). Long-acting nitrates have potential value in improving large arterial distensibility in patients with essential hypertension independent of blood pressure alteration. It might be used as an effectively additive drug in hypertension control. PMID- 11409657 TI - Ethnic and environmental differences in various markers of dietary intake and blood pressure among Chinese Han and three other minority peoples of China: results from the WHO Cardiovascular Diseases and Alimentary Comparison (CARDIAC) Study. AB - Our aim was to examine differences in dietary intake and blood pressure (BP) and their associations in four different ethnic Chinese populations, the Han, the Uygur, the Kazak and Tibetan subjects. This study used a sub-database of the Chinese sample of the WHO-Cardiovascular Diseases and Alimentary Comparison (CARDIAC) Study. The WHO-CARDIAC Study was a multicenter cross-sectional study. In each center, 100 men and 100 women aged 48-56 years were selected at random from the local population. Various markers of dietary intake and their relation with BP were studied. The results of the present study indicated the following. 1) There were significant differences in mean BP and prevalence rates of hypertension, with both being higher in the Kazak and Tibetan subjects than in Han and Uygur subjects. 2) The highest mean body mass index (BMI) was observed in the Kazak subjects, while the highest 24-h urinary sodium (Na) and sodium to potassium (Na/K) ratio excretion were observed in the Tibetan subjects. There were also significant differences in other factors, such as magnesium, 3 methylhistidine (3MH) (a biological marker of animal protein intake) and taurine (a biological marker of seafood intake) excretion levels among the four ethnic peoples. 3) In general, BMI, Na and/or Na/K ratios were positively, and 3MH/creatinine and taurine/creatinine ratios were negatively associated with BP. 4) After adjustment for age, sex and potassium, subjects with obesity (BMI > or =26 kg/m2) had significantly higher relative risk of being hypertensive (HT) than those with BMI<26 kg/m2 in the Han, Uygur and Kazak populations; and subjects with elevated sodium excretion (Na > or =244 mmol/day) had significantly higher relative risk of being HT than those with Na<244 mmol/day in the Han, Uygur and Tibetan populations. In conclusion, mean BP and prevalence rates of hypertension were significantly different among the four ethnic groups. These differences are likely to be due, at least in part, to the differences in several diet-related factors, which in turn are associated with culture and environmental differences. Different health promotion strategies might thus be emphasized in different populations. PMID- 11409658 TI - An angiotensin II type 1 receptor blocker, candesartan, increases myocardial apoptosis in rats with acute ischemia-reperfusion. AB - Angiotensin II (Ang II) and apoptosis contribute significantly to myocardial ischemia-reperfusion (I-R) injury. Evidence indicates that Ang II may activate apoptosis in myocytes. The present study was undertaken to investigate the effects of angiotensin receptor blockers (ARBs), candesartan, on the apoptosis of cardiac myocytes in rats after I-R. Rats were divided into a control group, a candesartan group I (0.015 mg/kg), and a candesartan group II (0.03 mg/kg). Candesartan was intravenously administered 30 min before ischemia. All rats were subjected to 30 min of coronary occlusion followed by 3 h of reperfusion. The data showed that left ventricular (LV) systolic pressure and LV +dp/dt was decreased after administration of candesartan, but increased after reperfusion in the candesartan group II, compared with those in the candesartan group I and control group. LV -dp/dt was decreased after candesartan administration in candesartan group II. The number of apoptotic cells in the candesartan groups (497+/-204 and 543+/-254, respectively) was higher than that in the control group (287+/-166; p<0.05). There was no significant difference in infarct size among the three groups. However, plasma CPK was lower in the candesartan groups than in the control group. Northern blot analysis showed that p53 mRNA was upregulated in the candesartan groups, in association with increased expression of bax mRNA. Immunohistochemical analysis showed that p53 and bax immunoreactivity were increased in both of the candesartan groups. In conclusion, candesartan increased apoptosis in the rat hearts after acute I-R, and this increase was possibly mediated by upregulation of p53 and bax gene expressions. In addition, candesartan was shown to improve LV function, in association with reduction of CPK release. PMID- 11409659 TI - COX-2 inhibition: an advance or only pharmaceutical "hype"? PMID- 11409660 TI - Identification of case complexity and increased health care utilization in patients with rheumatoid arthritis. AB - OBJECTIVES: To document biopsychosocial profiles of patients with rheumatoid arthritis (RA) by means of the INTERMED and to correlate the results with conventional methods of disease assessment and health care utilization. METHODS: Patients with RA (n = 75) were evaluated with the INTERMED, an instrument for assessing case complexity and care needs. Based on their INTERMED scores, patients were compared with regard to severity of illness, functional status, and health care utilization. RESULTS: In cluster analysis, a 2-cluster solution emerged, with about half of the patients characterized as complex. Complex patients scoring especially high in the psychosocial domain of the INTERMED were disabled significantly more often and took more psychotropic drugs. Although the 2 patient groups did not differ in severity of illness and functional status, complex patients rated their illness as more severe on subjective measures and on most items of the Medical Outcomes Study Short Form 36. Complex patients showed increased health care utilization despite a similar biologic profile. CONCLUSIONS: The INTERMED identified complex patients with increased health care utilization, provided meaningful and comprehensive patient information, and proved to be easy to implement and advantageous compared with conventional methods of disease assessment. Intervention studies will have to demonstrate whether management strategies based on INTERMED profiles can improve treatment response and outcome of complex patients. PMID- 11409661 TI - Use of alternative therapies by older adults with osteoarthritis. AB - OBJECTIVE: To examine the rates of use and expenditures on alternative therapies by adults with osteoarthritis (OA). METHODS: Adults with OA recruited from the community to participate in a randomized clinical trial recorded alternative and traditional health care use on postcard diaries. General and arthritis-specific quality of life was assessed by questionnaires. RESULTS: More than 47% of participants reported using at least one type of alternative care during the 20 week intervention period. Among alternative care consumers, the most commonly used treatments were massage therapy (57%), chiropractic services (20.7%), and nonprescribed alternative medications (17.2%). Four percent of subjects reported using only alternative care during the study period. Expenditures for alternative therapy averaged $1,127 per year, compared with $1,148 for traditional therapies. CONCLUSION: Use of and expenditures for alternative care were high among this cohort of older adults with OA. Clinicians may want to inquire about use of these therapies before recommending treatments for this condition. PMID- 11409662 TI - Primary care-based physical activity programs: effectiveness in sedentary older patients with osteoarthritis symptoms. AB - OBJECTIVE: This study examined, in a group of older patients, (a) the effectiveness of an invitation to participate in a program providing individualized physical activity advice in a primary care setting and (b) the changes in self-reported physical activity and symptoms in patients with osteoarthritis (OA). METHODS: Healthy, sedentary community-dwelling men and women aged 60 years or more were invited to participate. Following random allocation, the intervention group received individualized physical activity advice at baseline and at 3, 6, and 12 months followup. RESULTS: Of the 299 people who satisfied the study's inclusion criteria, a subgroup of 69 people reported pain and stiffness of the hip or knee at baseline. These patients reported increases in frequency and time of walking and vigorous exercise (all P < 0.001), with no change to OA symptom scores (pain and stiffness), and a small decline in physical functioning was reported at 12 months followup in the control group only (P = 0.027). At the 12-month followup more intervention participants than control participants (P = 0.013) reported a greater intention to exercise. CONCLUSIONS: An offer of primary care-based physical activity advice, with an emphasis on the benefits for general health (rather than "treatment" for OA), will attract individuals with OA symptoms. Although the present study was unable to demonstrate intervention-control group differences for the majority of outcomes, intention to exercise did appear to be positively influenced. PMID- 11409663 TI - Tendon disorders attributed to fluoroquinolones: a study on 42 spontaneous reports in the period 1988 to 1998. AB - OBJECTIVE: Fluoroquinolone antibiotics have been associated with tendinitis and tendon rupture. In this paper we report on the followup of 42 spontaneous reports of fluoroquinolone-associated tendon disorders. METHODS: This study is based on cases of fluoroquinolone-associated tendon disorders reported to the Netherlands Pharmacovigilance Foundation Lareb and the Drug Safety Unit of the Inspectorate for Health Care between January 1, 1988, and January 1, 1998. By means of a mailed questionnaire, we collected information on the site of injury, onset of symptoms, treatment, and course of the tendon disorder as well as information on possible risk factors and concomitant medication. RESULTS: Of 50 mailed questionnaires, 42 (84%) were returned. The data concerned 32 patients (76%) with tendinitis and 10 patients (24%) with a tendon rupture. Sixteen cases (38%) were attributed to ofloxacin, 13 (31%) to ciprofloxacin, 8 (19%) to norfloxacin, and 5 (12%) to pefloxacin. There was a male predominance, and the median age of the patients was 68 years. Most of the reports concerned the Achilles tendon, and 24 patients (57%) had bilateral tendinitis. The latency period between the start of treatment and the appearance of the first symptoms ranged from 1 to 510 days with a median of 6 days. Most patients recovered within 2 months after cessation of therapy, but 26% had not yet recovered at followup. CONCLUSION: These reports suggest that fluoroquinolone-associated tendon disorders are more common in patients over 60 years of age. Ofloxacin was implicated most frequently relative to the number of filled prescriptions in the Netherlands. PMID- 11409664 TI - The effectiveness and toxicity of cyclosporin A in rheumatoid arthritis: longitudinal analysis of a population-based registry. AB - OBJECTIVE: To determine factors associated with response or toxicity to cyclosporin A (CSA) in a population-based inception cohort with rheumatoid arthritis (RA). METHODS: Prospectively collected longitudinal measures including tender joint count (JC), duration of morning stiffness (MS), systolic and diastolic blood pressure (SBP, DBP), and serum creatinine (SCr) were modeled using generalized estimating equations. Survival methods were used to estimate CSA continuation time and its determinants. RESULTS: Of 133 patients (75% female, median RA duration 13 years), 37 discontinued CSA because of ineffectiveness (19) or because of toxicity (18) including increased SCr in 10, hypertension in 4, infections in 3, and gingival hyperplasia in 1. Patients remained on CSA a median of 75 months (95% confidence interval [CI] 38-112). Those receiving concomitant methotrexate (MTX) were more than 4 times as likely to continue on CSA therapy (hazard ratio 0.22, 95% CI 0.10-0.94). A lower final JC was predicted by a longer CSA treatment duration (relative risk [RR] 0.99 per month, 95% CI 0.98-0.99) and concomitant MTX therapy (RR 0.79, 95% CI 0.63-0.99); decreased MS was predicted only by longer CSA treatment duration (reduction of 2.0 minutes per month, 95% CI 1.1-3.0). Each previous disease-modifying antirheumatic drug (DMARD) exposure predicted a rise in SCr (35 micromole/liter, 95% CI 22-48), SBP (7.2 mm Hg, 95% CI 2.7-11.7), and DBP (3.8 mm Hg, 95% CI 3.0-6.4). CONCLUSIONS: Combination CSA/MTX prolongs therapy and reduces JC. Long-term CSA treatment was fairly well tolerated. Previous DMARD use appears to be a determinant for the development of toxicity. PMID- 11409665 TI - Testing the effectiveness of an osteoporosis educational program for nursing students in Thailand. AB - OBJECTIVE: Osteoporosis, a health problem that is on the rise, has received considerable attention among the health care community and the public. The majority of primary prevention programs for osteoporosis have been focused on women in mid-life. A concern is that young women may not be aware of osteoporosis risk factors and therefore may not be engaging in preventive behaviors. The purpose of this study was to test the effectiveness of an osteoporosis educational program for young women. METHODS: A sample of 100 female undergraduate students who were enrolled in the first year of a nursing program in Thailand were randomly assigned to a control group or an experimental group. Participants in the experimental group participated in a 3-hour osteoporosis educational program (OEP). At entry to and exit from the study, all participants completed the Osteoporosis Knowledge Test, the Osteoporosis Health Belief Scale, and the Osteoporosis Self-Efficacy Scale. RESULTS: The experimental group had higher change scores for knowledge, health belief, and self-efficacy than the control group. The OEP increased knowledge of osteoporosis in these young women. CONCLUSION: These findings indicate the need for further health education concerning the importance of dietary calcium and exercise on osteoporosis prevention in young women. PMID- 11409666 TI - Differential efficacy of tumor necrosis factor inhibition in the management of inflammatory eye disease and associated rheumatic disease. AB - OBJECTIVE: To evaluate the clinical usefulness of tumor necrosis factor (TNF) inhibitors in patients with inflammatory eye disease that is resistant to conventional immunosuppressive therapies. METHODS: Sixteen patients (4 males and 12 females aged 7 to 78 years) who received etanercept (n = 14) or infliximab (n = 2) for either inflammatory eye disease or associated joint disease were studied retrospectively to determine the effect of these medications on their ocular inflammation. RESULTS: Nine cases of uveitis and 7 cases of scleritis were treated. Systemic diagnoses included rheumatoid arthritis (n = 8), juvenile rheumatoid arthritis (n = 3), ankylosing spondylitis (n = 1), and psoriatic spondylarthropathy (n = 1). Three patients had uveitis without associated systemic disease. Although 12 of 12 patients with active articular inflammation (100%) experienced improvement in joint disease, only 6 of 16 with ocular inflammation (38%) experienced improvement in eye disease. Five patients developed inflammatory eye disease for the first time while taking a TNF inhibitor. No patient discontinued treatment because of adverse drug effects. CONCLUSION: TNF inhibitors are well tolerated immunosuppressive medications that may benefit certain subgroups of patients with inflammatory eye disease, but they appear to be more effective in controlling associated inflammatory arthritis. PMID- 11409667 TI - Comparative responsiveness of measures of pain and function after total hip replacement. AB - OBJECTIVE: To compare the responsiveness of the Functional Assessment System (FAS), the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), and the Medical Outcomes Study 36-item Short Form (SF-36) in patients with osteoarthritis (OA) scheduled for total hip replacement. METHOD: Twenty patients with a mean age at surgery of 72.6 years, with primary OA of the hip, were investigated preoperatively and at 3, 6, and 12 months postoperatively with the FAS, WOMAC, and SF-36. The responsiveness was calculated as standardized response mean, effect size, and relative efficiency. RESULTS: The pain and function scores of WOMAC and SF-36 showed greater responsiveness than FAS at 3 months. These differences remained at 6 and 12 months postoperatively. The differences between these 3 outcome measures were found to be similar using several methods for calculating responsiveness. CONCLUSION: Self-administered questionnaires like WOMAC and SF-36 are more responsive measures of pain and function than range of motion, performance tests, and observer-administered questions (FAS) following total hip replacement. PMID- 11409668 TI - Does satisfaction with abilities mediate the relationship between the impact of rheumatoid arthritis on valued activities and depressive symptoms? AB - OBJECTIVE: Both impact of rheumatoid arthritis (RA) on valued life activities and dissatisfaction with abilities have been linked to depression among individuals with RA. We integrated these concepts by examining the hypothesis that satisfaction with one's abilities may explain the mechanism by which the impact of RA on valued activities leads to depression. METHODS: Data were collected over 2 years (1997 and 1998) through interviews with the University of California, San Francisco, RA panel. Analyses examined whether activity impairment in 1997 predicted later (1998) dissatisfaction with abilities and depression. RESULTS: Greater impact on activities predicted dissatisfaction with abilities, which in turn was associated with higher depression scores. There was no direct relationship between activity impact and depression when satisfaction with abilities was considered. CONCLUSION: Satisfaction with abilities mediated the relationship between the impact of RA on valued activities and an increase in depressive symptoms, suggesting a need to assess not only physical decline but also individuals' interpretation of the decline. PMID- 11409669 TI - "Are you better?" A qualitative study of the meaning of recovery. AB - PURPOSE: Research into the meaning of illness has often focused on an individual's transition into a state of being ill, for example the adoption of a sick role. The question "Are you better?" addresses the transition out of this state and is fundamental to the patient-clinician relationship, guiding decisions about treatment. However, the question assumes that all patients have the same meaning for "being better." The purpose of this study was to explore the meaning of the concept of recovery (getting better) in a group of people with upper limb musculoskeletal disorders. METHODS: Qualitative (grounded theory) methods were used. Individual interviews were conducted with 24 workers with work-related musculoskeletal disorders of the upper limb. The audiotaped interviews were transcribed and coded for content. Categories were linked, comparisons made, and a theory built about how people respond to the question "Are you better?" RESULTS: The perception of "being better" is highly contextualized in the experience of the individual. Being better is not only reflected in changes in the state of the disorder (resolution) but could be an adjustment of life to work around the disorder (readjustment) or an adaptation to living with the disorder (redefinition). The experience of the disorder can be influenced by factors such as the perceived legitimacy of the disorder, the comparators used to define health and illness, and coping styles, which in turn can influence being better. CONCLUSION: Two patients could mean very different things when saying that they are better. Some may not actually have a change in disease state as measured by symptoms, impairments, or function. PMID- 11409670 TI - Rheumatology outpatient nurse clinics: a valuable addition? AB - OBJECTIVES: "Transmural rheumatology nurse clinics," where nursing care is provided under the joint responsibility of a home care organization and a hospital, were recently introduced into Dutch health care. This article gives insight into outcomes of the transmural rheumatology nurse clinics. METHODS: Patients with rheumatologic conditions who attended a transmural nurse clinic, in addition to receiving regular care, were compared with patients with rheumatologic conditions who received regular care only. The main outcome measures were the need for rheumatology-related information, the use of aids and adaptations, the use of health care services, and daily functioning. RESULTS: Attending a transmural nurse clinic does not influence patients' need for information, the application of practical aids and adaptations, or daily functioning. However, attending a transmural nurse clinic does result in more contacts with rheumatologists and occupational therapists. CONCLUSIONS: Attending transmural nurse clinics does not result in major differences in outcomes compared with regular care. Further studies are needed to appreciate the long term effects of transmural nurse clinics. PMID- 11409671 TI - Walking velocity in aged persons: its association with lower extremity joint range of motion. AB - OBJECTIVE: To measure the association between walking velocity and hip and knee flexion range, ethnic background, anthropometrics, comorbid pathologies, and coimpairments, in a sample of community-dwelling aged persons. METHODS: To reach our objective, we used data from the San Antonio Longitudinal Study of Aging (SALSA), a population-based cohort of Mexican American and European American persons aged 64 to 79. By fitting hierarchical regression models, we measured the variance in the walking velocity over 50 feet explained by hip and knee flexion range, adjusting for the combined influence of demographic and anthropometric variables, coexistent pathologic conditions, impairments, and the examiners who conducted the assessments. RESULTS: The average walking velocity among the 702 subjects for whom data were available was 73.6 meters per minute (range 20 to 121). Bivariate analyses revealed significant associations between walking velocity and age, sex, ethnic background, height, weight, presence of arthritis, diabetes mellitus, stroke, upper leg pain, peripheral vascular disease, left ventricular hypertrophy, and forced expiratory volume at 1 second. The correlation (r) between walking velocity and flexion range of the hip and knee were 0.40 and 0.35, respectively (P < or = 0.001 for each). Multivariate hierarchical models adjusting for demographic and anthropometric characteristics of the subjects, and examiner variation, revealed that hip and knee flexion range explained 6% of the variance in walking velocity. Adjusting for the presence of comorbid conditions and coimpairments reduced the variance attributable to hip and knee flexion range only slightly, to 5%. CONCLUSION: Hip and knee flexion range contribute significantly to walking velocity in the SALSA cohort of community-dwelling aged persons. PMID- 11409672 TI - Pre-appointment management of new patient referrals in rheumatology: a key strategy for improving health care delivery. AB - OBJECTIVE: To analyze the impact of a rheumatologist reviewing each newly referred patient's medical records prior to scheduling an appointment (pre appointment management). METHODS: All 279 new patients who were referred in the 6 months after initiating pre-appointment management were studied. The authors reviewed systemwide patient records, appointment intake information, visit schedules, physician comments, and patient complaint data. RESULTS: Only 59% of referred patients required a rheumatology consultation for appropriate care. Some problems were rapidly resolved without consultation. In some cases, other specialty consultation or continuing prior care was considered to be more appropriate. The latter alternative did not compromise these patients' outcomes. Practice access and efficiency were improved. Profitability was maintained. Referring physicians and patients were generally accepting and cooperative. CONCLUSION: New patient pre-appointment management should be a key strategy for reducing health care costs, addressing personnel shortage, and improving access to and coordination of rheumatic disease care. PMID- 11409673 TI - Living with osteoarthritis: patient expenditures, health status, and social impact. AB - OBJECTIVE: To determine "out-of-pocket" expenditures related to osteoarthritis (OA) and to explore whether demographic details, health status scores (Medical Outcomes Study 36-item Short Form [SF-36] and Western Ontario and McMaster Universities Osteoarthritis Index [WOMAC]), or perception of social effect were expenditure determinants. METHODS: A prospective cohort study of community dwelling subjects with OA completed 4 consecutive 3-month cost diaries. In addition, subjects completed the SF-36 and WOMAC at baseline and at 12 months. Social impact at baseline was collected. Four groups categorized by age and sex were compared. Patients undergoing joint replacement were excluded. RESULTS: Differences in health status were defined more by age than by sex, especially for physical function. The costs to the patients were high, particularly for women, who spent more on medications and special equipment. Women also reported receiving more assistance from family and friends. Higher disease-related expenditures were associated with greater pain levels, poorer social function and mental health, and longer duration of disease. Significant independent predictors of total patient expenditures related to OA were being female and having joint stiffness. CONCLUSION: Despite having heavily subsidized health care and access to the Pharmaceutical Benefits Scheme, out-of-pocket costs for patients with OA in Australia are considerable. Higher expenditures for patients with OA are related to more advanced disease, especially for women. PMID- 11409674 TI - Oral health promotion evaluation--time for development. AB - Increasing emphasis is now being placed upon the evaluation of health service interventions to demonstrate their effects. A series of effectiveness reviews of the oral health education and promotion literature has demonstrated that many of these interventions are poorly and inadequately evaluated. It is therefore difficult to determine the effectiveness of many interventions. Based upon developments from the field of health promotion research this paper explores options for improving the quality of oral health promotion evaluation. It is essential that the methods and measures used in the evaluation of oral health promotion are appropriate to the intervention. For many oral health promotion interventions clinical measures and methods of evaluation may not be appropriate. This paper outlines an evaluation framework which can be used to assess the range of effects of oral health promotion programmes. Improving the quality of oral health promotion evaluation is a shared responsibility between researchers and those involved in the provision of programmes. The provision of adequate resources and training are essential requirements for this to be successfully achieved. PMID- 11409675 TI - Social inequality in oral health and use of dental care in Sweden. AB - OBJECTIVE: To describe oral health and use of dental care in relation to socio economic determinants over time in Sweden. METHODS: Cross-sectional study based on interview data on two randomly sampled sequential populations consisting of 7,610 Swedish adult (25-64 years) residents and 4,315 children (3-15 years) in their households from the Survey of Living Conditions 1996-97, and 7,649 adult Swedish residents (25-64 years) from the survey of 1988-89. RESULTS: Low educational level, having no cash margin and being born outside of Sweden was associated with higher odds of problems with chewing, wearing a prosthesis and not having been treated by a dentist during the 24 months preceding the interview, in a logistic regression analysis of data from the 1996-97 survey in the adult study population (adjusted odds ratios 1.6-2.9). The same socio economic determinants were associated with caries in children (adjusted odds ratios 1.2-1.5). The socio-economic differences in dental treatment and problems with chewing were greater in the age group 45-64 years compared to 25-44-year olds. The prevalence of problems with chewing increased from 7.1% (95% CI 6.5 8.1) in the 1988-89 survey to 9.1% (8.4-9.8) in the 1996-97 survey. A similar increase, from 2.4% (2.2-2.6) to 4.4% (3.9-4.9) was observed for individuals not having been in dental treatment during the last 24 months. The socio-economic distribution of oral health and use of dental care in the adult population was similar in the two surveys. CONCLUSION: This study demonstrates that socio economic differences in oral health and use of dental care are most marked in older (45-64 years) adults in Sweden, but are significant in young adults and, in terms of oral health, in children as well. A steep increase in user charges during the 1990s has been paralleled by a moderate increase in problems with chewing and the proportion of the population that has no regular dental care, which suggests a link that needs to be evaluated in further studies. PMID- 11409676 TI - An assessment of sensitivity to change of the Oral Health Impact Profile in a clinical trial. AB - Patient-based assessment of oral health outcomes is of growing interest. Measurement of change following clinical intervention is a key property of a health status measure. To date, most of the research on oral health status measurement has focused on construct and discriminant validity of health status measures. OBJECTIVES: The objective of this study was to assess sensitivity to change of an oral-specific health status measure, the Oral Health Impact Profile (OHIP). METHODS: Study subjects were in three groups, namely, edentulous/edentate subjects who requested and received complete implant stabilised oral prostheses (IG, n=26), edentulous/edentate subjects who requested implants but received conventional dentures (CDG1, n=22), and edentulous subjects who had new conventional complete dentures (CDG2, n=35). Data were collected pre- and post operatively using the OHIP and a validated denture satisfaction questionnaire. RESULTS: All subjects reported similar low levels of denture satisfaction pre operatively. Denture problems had a more significant impact on oral health related quality of life (OHRQL) for implant seekers (IG and CDG1 subjects) than subjects seeking conventional dentures (CDG2). Following treatment, significant improvement in satisfaction with oral prostheses and OHRQL was reported by IG and CDG2 subjects; the level of improvement was more moderate for CDG1 subjects. OHIP change scores were correlated with denture satisfaction change scores. CONCLUSIONS: It was concluded that sensitivity to change of the OHIP was good. This property was not improved by using statement weights. PMID- 11409677 TI - Type 1 diabetes mellitus and oral health: assessment of coronal and root caries. AB - OBJECTIVES: The oral health of a large cohort of adult insulin-dependent diabetic patients (Type 1), diagnosed 24 years previously with juvenile onset, was comprehensively assessed. This paper describes the prevalence of coronal and root caries in this adult Type 1 diabetic population and evaluates demographic, dietary, behavioral, physiologic, salivary and medical variables associated with decayed and filled surfaces in the crown (DFS) or root (RDFS). METHODS: Type 1 diabetes mellitus subjects participating in this oral health evaluation had been monitored for 6-8 years as participants in the University of Pittsburgh, Department of Epidemiology, longitudinal study of medical complications associated with diabetes. Four hundred and six diabetic subjects received a comprehensive oral health examination during one of their regularly scheduled medical visits. Oral assessments included coronal and root caries, missing teeth, edentulism, periodontal status, soft tissue pathologies, salivary function and health behaviors. Sixteen diabetic subjects and one control subject were edentulous. Coronal and root caries data from the remaining 390 dentate diabetic subjects were compared with 202 dentate nondiabetic control subjects. RESULTS: The adult Type 1 diabetic subjects were not found to have significantly higher DFS rates as compared with our control subjects or published age-adjusted NHANES III findings. Both control and diabetic subjects had low decayed to filled tooth surface ratios. A linear regression model evaluated possible associations with coronal decayed and filled tooth surfaces (DFS) within the diabetic population. Significant factors included older age, women, fewer missing teeth, more frequent use of dental floss, more frequent visits to the dentist during the last 12 months, and diabetic nephropathy. The prevalence of RDFS was higher in the diabetic subjects as compared to recruited control subjects. Neither dietary behaviors nor glycemic control were found to contribute to coronal or root caries. CONCLUSIONS: Factors associated with presence of coronal and root caries and fillings are discussed. Possible causes and implications for the association between DFS and diabetic nephropathy are provided. PMID- 11409678 TI - Prevalence of impacts of dental and oral disorders and their effects on eating among older people; a national survey in Great Britain. AB - OBJECTIVE: The objective was to assess the prevalence, in a British population aged 65 years and older, of oral health related impacts and the effects they had on the quality of daily life and in particular on eating. METHODS: 753 free living and 202 institutionalised subjects aged 65 years and over, participating in the oral health survey of the British National Diet and Nutrition Survey (NDNS), had a dental examination and interview. Data on the impact of dental and oral disorders on the activities of daily living based upon the modified Oral Impacts on Daily Performance (OIDP) indicator were collected. RESULTS: 17% of the free living edentate participants reported that their mouth affected their pattern of daily living on a regular basis. Oral impacts levels were lowest in dentate subjects with the greatest number of teeth. For the dentate, the most common oral impacts were on eating and speaking. Impacts relating to emotional stability, sleeping, relaxing, carrying out physical activity and social contact were very infrequent, but were severe when they did occur. Among those with an impact on eating, 25% said it was severe and 42% had the impact nearly every day or in a spell of 3 or more months. Oral impacts were more prevalent among the institution sample, particularly the dentate. The impacts were associated with the inability or difficulty to eat a range of 16 common foods. CONCLUSION: This survey has shown that the oral status of older people fairly frequently affects the quality of life of older people, and in particular, the ability to eat several common types of foods. PMID- 11409679 TI - Adolescents' sense of coherence, oral health status, and oral health-related behaviours. AB - OBJECTIVE: To investigate the relationship between sense of coherence (SOC) and oral health. It was hypothesised that subjects with better oral health status and better oral health-related behaviours have higher levels of SOC. METHODS: A cross sectional study was conducted in Goiania-GO, Middle-West Brazil, on a sample of 664 15-year-olds randomly selected from schools. Data were collected through questionnaires, the short version of Antonovsky's SOC Scale (13-item) and clinical dental examinations. Multiple logistic regression and polytomous ordered regression were used in the data analysis. Two sets of outcome variables were selected for the analyses: oral health status (dental caries, oral cleanliness, and periodontal disease), and oral health-related behaviours (frequency of sugar intake, toothbrushing frequency, and pattern of dental attendance). RESULTS: Adolescents' SOC was associated with their caries experience in anterior teeth (OR=0.81 for 10 units increase in SOC scale; 95% CI=0.66-0.98), but the relationship did not remain significant after controlling for other factors. Adolescents with higher SOC were less likely to visit the dentist mainly when in trouble, compared with those with lower SOC (OR=0.83, 95% CI=0.71-0.98), or equivalently more likely to visit for mainly check-ups. Other measures of oral health status and behaviours were not significantly associated with SOC. CONCLUSION: SOC was identified as a psychosocial determinant of adolescents' oral health-related behaviour, particularly affecting their pattern of dental attendance. PMID- 11409680 TI - Evaluation of oral health promotion in the workplace: the effects on dental care costs and frequency of dental visits. AB - An oral health promotion program has been conducted since 1989 at a shipyard in Japan. AIMS: The purpose of the present study was to assess the impact of oral health promotion in the workplace in terms of dental care costs and frequency of dental visits. METHODS: This program consisted of orientation, an initial regimen, and group counseling. The initial regimen included an evaluation of each participant's oral health status, and instruction concerning oral hygiene, and prevention of oral disease. The participants were selected with each unit being a preexisting peer group. Eighty-seven participants were compared with 216 control subjects in terms of annual dental care costs and frequency of dental visits in four periods: the 1-year period before the program, the 1st year after the program, the 2nd year after, and the 3rd year after. RESULTS: The annual mean dental care cost in the participant group for the year prior to the program was higher than that in the control group (21,317 vs. 17,116 yen). In the 1st year after the program, the difference increased (26,642 vs. 19,481 yen). In the 2nd and 3rd years after the program, dental care costs in the participant group were lower than those in the control group (2nd year: 18,305 vs. 22,841 yen, 3rd year: 16,911 vs. 21,920 yen). CONCLUSIONS: It can be concluded that this oral health promotion program in the workplace contributed to saving of costs associated with dental care. The workplace can be regarded as a key area for implementation of an oral health care system to make good use of limited resources. PMID- 11409681 TI - Modified IOTN: an orthodontic treatment need index for use in oral health surveys. AB - This paper reports the results of a study of the reliability of a modified version of the Index of Orthodontic Treatment Need (IOTN) for use in oral health surveys. Twelve non-specialist dental examiners were trained in the use of the Modified IOTN using a standardised teaching protocol lasting approximately 1.5 hours. Following a school-based calibration exercise it was found that nearly all the examiners achieved either good or excellent agreement (mean Kappa=0.74). The average sensitivity and specificity scores were 0.90 and 0.84, respectively. The Modified IOTN appears to overcome the training and reliability problems that often accompany the use of orthodontic indices by non-specialists in oral health surveys. PMID- 11409682 TI - Socioeconomic factors that affect international caries levels. AB - Health policy makers have examined the impact of economic policies and a nation's social development on the health of its population. OBJECTIVES: The purpose of this research was to investigate the association between health care expenditures, socioeconomic factors, and caries levels in 109 countries representing an array of social and economic conditions, and to determine how such factors are related to caries experience at different levels of economic development. METHODS: Countries were divided into established market economies and three groups of developing nations, based on their GNP (high, medium, low income). Total health expenditures as percent GDP and as US dollars per capita, public expenditures and aid flow as percent of total health expenditures, dentists per 100,000 population, per capita sugar consumption, and urbanization of the population were compared among these countries. Correlations between these variables, and decayed, missing and filled teeth (DMFT) of 12-year-olds in these countries were assessed. RESULTS: The highest correlations were found between DMFT and public expenditures, sugar consumption, and urbanization. Highly significant differences emerged across nation groups by stages of development on several variables. CONCLUSIONS: The findings suggest that it is important to consider the stage of development of a given country when planning caries prevention policies and programs. Such a contextual approach is more likely to be successful in reducing caries levels. PMID- 11409683 TI - Rugby players' satisfaction with custom-fitted mouthguards made with different materials. AB - OBJECTIVES: This study compared the comfort of two bimaxillary custom-fitted mouthguards. One type was made with silicone rubber and the other with methyl methacrylate (acrylic). METHODS: The study was a within-subject crossover clinical trial with 52 high-school rugby players who were randomly allocated to one of two groups. The first group wore a silicone mouthguard for 4 months and an acrylic one for the following 4-month period. The second group wore an acrylic mouthguard followed by a silicone one for similar periods. Comfort, bulkiness, stability, hardness, ability to talk and to breathe, oral dryness, nausea and inclination to chew were evaluated for each period using a Visual Analogue Scale questionnaire. RESULTS: There was no significant difference concerning comfort, bulkiness, ability to talk and to breathe, oral dryness and nausea between silicone and acrylic mouthguards by group and time of examination (Three-way ANOVA P>0.05). Acrylic mouthguards were more stable and harder than silicone ones (Wilcoxon's test P<0.01). Tendency to chew was greater for silicone appliances (P<0.01). For stability, hardness and inclination to chew, there was no significant difference in the response of the players based on the sequence of use of the two types of mouthguard during the survey (Mann-Whitney test P>0.05). At the end of the study, 56% of the players preferred to keep the acrylic mouthguard and 44% chose the silicone one. This choice did not vary between the groups (chi2, P>0.05). CONCLUSION: Silicone rubber mouthguards were well accepted by the players but technical improvements in silicone materials are needed to improve hardness and stability of silicone mouthguards for sport. PMID- 11409684 TI - Impairment of antigen-specific cellular immune responses under simulated microgravity conditions. AB - Microgravity has been implicated to play a role in the observed immune dysfunction of astronauts and cosmonauts after either short-term or long-term space travel. These reports, together with studies describing increased levels of microorganisms in the space cabin environment suggest potential risk for in flight incidences of infectious diseases. In order to understand the mechanism underlying these immune defects, it is important to have a ground-based model that would reliably mimic the effects of microgravity on antigen-specific immune function. We tested the utility of the rotating wall vessel (RWV) technology developed at NASA as a model system because in the RWV the culture medium and the cells rotate synchronously with the vessel, thereby creating simulated microgravity conditions. We compared the RWV to the conventional tissue culture flask (T-flask), for culturing immune precursor cells with cytotoxic T lymphocyte (CTL) activity against synthetic viral peptides. We observed a significant loss of antigen-specific CTL activity in RWV cultures, but not in those from the T flask, irrespective of the peptide immunogen used for inducing the primary immune response in different mouse strains. Loss of CTL activity in RWV cultures coincided with a significant reduction in CD8+ cells as well as CD4+ cells and DEC205+ dendritic cells, suggesting adverse effects of RWV culturing on both the effector and accessory cells for the loss of antigen-specific CTL function. These results provide a strong parallel to the reported defects in cell-mediated immunity during space travel and strongly support the utility of the RWV technology as an effective ground-based model for identifying key steps in immune cell dysfunction related to microgravity. PMID- 11409685 TI - Simulated microgravity impairs respiratory burst activity in human promyelocytic cells. AB - The concept of microgravity (free-fall) influencing cellular functions in nonadherent cells has not been a part of mainstream scientific thought. Utilizing rotating wall vessels (RWVs) to generate simulated microgravity conditions, we found that respiratory burst activity was significantly altered in nonadherent promyelocytic (HL-60) cells. Specifically, HL-60 cells in simulated microgravity for 6, 19, 42, 47, and 49 d had 3.8-fold fewer cells that were able to participate in respiratory burst activity than cells from 1 x g cultures (P = 0.0011, N = 5). The quantity of respiratory burst products from the cells in simulated microgravity was also significantly reduced. The fold increase over controls in mean fluorescence intensities for oxidative products from cells in microgravity was 1.1+/-0.1 versus 1.8+/-0.3 for cells at 1 x g (P = 0.013, N = 4). Furthermore, the kinetic response for phorbol ester-stimulated burst activity was affected by simulated microgravity. These results demonstrate that simulated microgravity alters an innate cellular function (burst activity). If respiratory burst activity is impaired by true microgravity, then recovery from infections during spaceflight could be delayed. Finally, RWVs provide an excellent model for investigating the mechanisms associated with microgravity-induced changes in nonadherent cells. PMID- 11409686 TI - Characteristics of human dendritic cells generated in a microgravity analog culture system. AB - Generation of an effective immune response requires that antigens be processed and presented to T lymphocytes by antigen-presenting cells, the most efficient of which are dendritic cells (DC). Because of their influence on both the innate and the acquired arms of immunity, a defect in DC would be expected to result in a broad impairment of immune function, not unlike that observed in astronauts during or after space flight. In the study reported here, we investigated whether DC generation and function are altered in a culture environment that models microgravity, i.e., the rotary-cell culture system (RCCS). We observed that RCCS supported the generation of DC identified by morphology, phenotype (HLA-DR+ and lacking lineage-associated markers), and function (high allostimulatory activity). However, the yield of DC from RCCS was significantly lower than that from static cultures. RCCS-generated DC were less able to phagocytose Aspergillus fumigatus conidia and expressed a lower density of surface HLA-DR. The proportion of DC expressing CD80 was also significantly reduced in RCCS compared to static cultures. When exposed to fungal antigens, RCCS-generated DC produced lower levels of interleukin-12 and failed to upregulate some costimulatory/adhesion molecules involved in antigen presentation. These data suggest that DC generation, and some functions needed to mount an effective immune response to pathogens, may be disturbed in the microgravity environment of space. PMID- 11409687 TI - Epstein-Barr virus latently infected cells are selectively deleted in simulated microgravity cultures. AB - Rotating-wall vessels (RWVs) allow for the cultivation of cells in simulated microgravity. Previously, we showed that the cultivation of lymphoblastoid cells in simulated microgravity results in the suppression of Epstein-Barr virus (EBV) reactivation. To determine if the suppression generated by simulated microgravity could be reversed by changing to static culture conditions, cells were cultured in an RRWV for 5 d, and then switched to static conditions. Following the switch to static conditions, viral reactivation remained suppressed (significantly lower) relative to static control cultures over a 4-d period. Additionally, experiments were conducted to determine if chemical treatment could induce viral reactivation in cells from simulated-microgravity cultures. Cells were cultured in static flask cultures and in simulated microgravity in RWVs for 4-7 d. The cells were then transferred to 50-cm3 tubes, and treated with 3 mM n-butyrate for 48 h, or 18 ng/ml of phorbol ester, viz., 12-0-tetradecanoylphorbol-13 acetate (TPA) for either 2 or 48 h, under static conditions. Although EBV was inducible, the cells from simulated-microgravity cultures treated with n-butyrate displayed significantly lower levels of viral-antigen expression compared with the treated cells from static cultures. Also, incubation with TPA for 2-3 h, but not for 48 h, reactivated EBV in cells from RWV cultures. In contrast, EBV was inducible in cells from static cultures treated for either 2-3 or 48 h with TPA. TPA reactivation of EBV following a 2-3-h period of treatment indicates that the protein kinase C signal-transduction pathway is not impaired in lymphoblastoid cells cultured in simulated microgravity. However, the exposure of B lymphoblastoid cells from simulated-microgravity cultures to TPA for more than 3 4 h triggered a lytic event (apoptosis or necrosis), which prevented replication of the virus. Thus, EBV-infected cells in simulated microgravity were negatively selected in the absence of any cytotoxic cells. PMID- 11409688 TI - Normal human liver organ culture. PMID- 11409689 TI - Human keratinocytes express functional alpha-MSH (MC1-R) receptors. PMID- 11409690 TI - Cryopreservation of heart cells from the eastern oyster. AB - Conditions were developed to cryopreserve cells from pronase-dissociated atria and ventricles of eastern oysters (Crassostrea virginica). The effect of three concentrations (5, 10, 15%) of the cryoprotectants (dimethyl sulfoxide, glycerol, and propylene glycol), three thawing temperatures (25, 45, 75 degrees C), and three cooling rates (slow, medium, fast) were compared. Cells were frozen at -80 degrees C and plunged in liquid nitrogen. Thawed cells were seeded in 96-well plates and primary cultures were evaluated after 3 d by measuring the metabolic activity using a tetrazolium compound, 3-(4,5-dimethylthiazol-2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, and by comparing the relative spreading of cells between treatments. The best conditions for freezing and thawing of cells for each cryoprotectant were selected and a final study was performed to compare cryoprotectants. For this final study, we measured the number of cells and their viability 3 d after thawing, in addition to determining cell metabolic activity and cell spreading. Primary cultures of cells fozen without cryoprotectant and of nonfrozen cells were used as controls in all studies. Atrial cells were best cryopreserved with glycerol at a concentration of 10%, a medium cooling rate, and thawing at 45 degrees C. After thawing, atrial cells showed 53+/-5% of the metabolic activity, 84+/-5% of the number, and 92+/ 2% of the viability of nonfrozen cells. For ventricular cells, 10% glycerol, a medium cooling rate, and thawing at 25 degrees C yielded the best results. The thawed ventricular cells showed 83+/-5% of the metabolic activity, 91+/-5% of the number, and 96+/-2% of the viability of nonfrozen cells. PMID- 11409691 TI - Transforming growth factor beta may act as an autocrine-survival-promoting factor for transformed trophoblasts. AB - Using five trophoblast cell lines of different differentiation status, we have shown that trophoblasts could constitutively release the transforming growth factor beta-1 (TGFbeta1), but not TGFbeta2. Treatment of the human tumorigenic, TL, and BeWo cells with a differentiating agent and a potent protein kinase C activator--the tumor-promoting agent--or the JEG-3 cells with cholera toxin--a potent cyclic adenosine 3':5'monophosphate (cAMP) inducer--or its analogue 8 bromo-cAMP, potentiates TGFbeta production, but the two signaling pathways appear to be mutually exclusive. Surprisingly, the JAR tell line failed to respond to either type of TGFbeta activator. Based on reverse transcriptase (RT)-polymerase chain reaction (PCR), it was found that only the JAR cell line expressed messenger ribonucleic acid for decorin, a natural inhibitor of TGFbeta, and none of the cell lines had detectable protein expression as determined by immunocytochemical studies. The tell number in cultures with decorin was invariably lesser than in those without decorin under serum-free conditions for all the cell lines tested. These results suggest that TGFbeta may act as an autocrine-survival factor for transformed trophoblasts by allowing the cells to survive longer under a microenvironment which is not favorable for growth. Lastly, our results indicate that decorin, acting in a paracrine manner, may also play an important negative regulatory role in the development of transformed trophoblasts by sequestering TGFbeta, thereby preventing its action. PMID- 11409693 TI - Park City, Utah Molecular Psychiatry Meeting, February 2001. PMID- 11409692 TI - Renal cell cultures for the study of growth factor interactions underlying kidney organogenesis. AB - The present study was performed in four renal cell lines to evaluate their capability to: (1) produce and express transforming growth factor alpha (TGFalpha), its respective receptor, the epidermal growth factor receptor (EGFr) and the small G protein, RhoA, and (2) exhibit morphogenetic properties when grown on Matri-cell substrates. The cell lines were derived from normal (Madin Darby canine kidney cells), embryonic (SK-NEP-1 and 293 cells), and cancerous (human renal adenocarcinoma cells) kidneys. TGFalpha messenger ribonucleic acid, evaluated by a nonradioactive in situ hybridization technique, was found to be expressed in all the cell lines. Large amounts of TGFalpha peptide were observed in all four cell lines, while EGFr was highly expressed only in cancerous ACHN and embryonic-tumor SK-NEP-1 cells. RhoA peptide was found in appreciable amounts in SK-NEP-1 and 293 cells (compared to the other two cell lines). The morphogenetic properties of the four cell lines were assessed, by culturing them on Matri-cell dishes: SK-NEP-1 cells alone were found to grow in three dimensional structures forming clusters and worm-like cellular aggregates. This feature was displayed by SK-NEP-1 cells but not by the other three cell lines, and may be connected with the contemporary presence of RhoA, EGFr, and TGFalpha found in significant amounts only in the SK-NEP-1 cell line. PMID- 11409694 TI - No association between serotonin 2A receptor gene variants and personality traits. AB - Among theories of biological underpinnings to personality traits, different mechanisms of the serotonergic system are perhaps the most common factors suggested to influence individual differences in personality traits. We have investigated two frequent variants in the serotonin 2A receptor gene (5-HT2A) and personality traits. Healthy Swedish subjects (n = 304) were assessed with the Karolinska Scales of Personality (KSP) inventory. After correction for multiple testing, no significant differences were found. We conclude that the investigated 5-HT2A gene variants do not significantly influence personality as assessed by the KSP in the present population. PMID- 11409695 TI - Androgen receptor trinucleotide repeat polymorphism and personality traits. AB - Human family and twin studies have established considerable heritable components influencing individual differences in personality traits as assessed by self report questionnaires. We have investigated a trinucleotide repeat polymorphism in the androgen receptor gene and personality traits. Healthy Swedish subjects (n = 335) were assessed with the Karolinska Scales of Personality inventory. There were tendencies (P > or = 0.006) in some scales indicating possible relationships between the androgen receptor allele length and personality traits related to dominance and aggression. However, after correction for multiple testing, no significant differences were found. We conclude that no significant association could be found between the androgen receptor polymorphism investigated and any personality trait, although the tendencies found are worthwhile subjects for replication attempts. PMID- 11409696 TI - Association of DRD4 with attention problems in normal childhood development. AB - Several previous studies found an association of clinically diagnosed attention deficit hyperactivity disorder with long alleles of a variation in the DRD4 dopamine receptor gene exon III coding sequence. We evaluated the DRD4 polymorphism in a non-clinically selected sample of children for whom maternal reports of attention problems were available at 4 and 7 years of age. There was a significant elevation in attention problem scores in children carrying DRD4 long alleles that accounted for 3-4% of total variation at each age and for 5-7% of the temporally stable component of the phenotype. Our results show that the DRD4 gene influences normal as well as pathological attention processes, and the results highlight the utility of longitudinal measurements in psychiatric genetics. PMID- 11409697 TI - DRD4 related to infant attention and information processing: a developmental link to ADHD? AB - The dopamine D4 receptor (DRD4) exon III polymorphism has generated interest because of its association with attention deficit hyperactivity disorder (ADHD), with an increased frequency of the seven-repeat allele being reported in children with ADHD. Deficits in sustained attention and information processing characterize ADHD, and individual differences in these functions are apparent from infancy. We found that in a structured play situation and on an information processing task, 1-year-old infants with the 7-DRD4 allele showed less sustained attention and novelty preference than do infants without the 7-DRD4 allele. There was also a significant interaction between DRD4 and the serotonin transporter promoter (5-HTTLPR) gene on a measure of sustained attention. Our results provide evidence for a possible developmental link between DRD4 and ADHD via early sustained attention and information processing. It also points to the importance of considering the influence of more than one gene in studies of behavior. PMID- 11409698 TI - Nicotinic acetylcholine receptor alpha4 subunit gene polymorphism and attention deficit hyperactivity disorder. AB - Attention deficit hyperactivity disorder (ADHD) is a highly heritable, common psychiatric disorder that presents in childhood and that probably involves several genes. There are several lines of evidence suggesting that the nicotinic system may be functionally significant in ADHD: (a) nicotine promotes the release of dopamine and has been shown to improve attention in adults with ADHD, smokers and non-smokers; (b) ADHD is a significant risk factor for early initiation of cigarette smoking in children; (c) maternal cigarette smoking appears to be a risk factor for ADHD; (d) animal studies in rats and monkeys also suggest that nicotine may be involved in attentional systems and locomotor activity; and (e) a central nicotinic agonist, ABT-418, improves attention in both monkeys and ADHD adults. The current study examined the alpha 4 receptor, one of the sites of action of ABT-418. A known Cfol polymorphism within the nicotinic acetylcholine alpha 4 receptor gene, CHRNA4, was studied in 70 ADHD parent-proband trios from an ongoing sample collection of children aged 6-12 with ADHD, according to DSM-IV criteria. Children with known major medical or psychiatric conditions or mental retardation (IQ < 70) were excluded from the study. The Transmission Disequilibrium Test demonstrated no evidence that variation at the nicotinic acetylcholine alpha 4 receptor Cfol polymorphism influences susceptibility to ADHD (P > 0.35). The continuing sample collection will enable further study of other potential nicotinic system polymorphisms in ADHD in more powerful samples. PMID- 11409699 TI - The phospholipase C-gamma1 gene (PLCG1) and lithium-responsive bipolar disorder: re-examination of an intronic dinucleotide repeat polymorphism. AB - Twin, family and adoption studies have indicated that genetic susceptibility plays an important role in the etiology of bipolar disorder. Turecki et al. (1998) recently published preliminary evidence suggesting that bipolar patients with an excellent response to lithium treatment have a higher frequency of a specific dinucleotide repeat allele in the phospholipase Cgamma-1 (PLCG1) genomic region. The present work was undertaken to re-examine the finding by Turecki et al. in a sample of Norwegian lithium-treated bipolar patients sub-classified as lithium responders, non-responders, or partial responders/unclassified. The overall distribution of the PLCG1 dinucleotide repeat alleles was not significantly different between different categories of subjects. When analyzed according to presence or absence of different dinucleotide alleles, a PLCG1-8 repeat was more frequent among lithium responders vs controls. In line with Turecki et al., we also noticed a moderately over-representation of the PLCG1-5 repeat among the bipolar patients as compared to the controls. PMID- 11409700 TI - Systematic screening for mutations in the glycine receptor alpha2 subunit gene (GLRA2) in patients with schizophrenia and other psychiatric diseases. AB - The glycine receptor, which is a member of the ligand-gated ion channel superfamily, mediates synaptic inhibition in the spinal cord and other brain regions. This superfamily has been implicated in the pathogenesis of schizophrenia and other psychiatric diseases. The complete coding sequence and splice junctions of the GLRA2 gene were scanned by DOVAM-S, a form of SSCP analysis with sufficient redundancy to detect virtually all mutations. Those analyses were performed in 113 patients with schizophrenia, and in pilot studies of patients with bipolar illness, alcoholism, puerperal psychosis, autism, and attention-deficit hyperactivity disorder (533 kb total scanned sequences). We detected three sequence changes in the coding region, all resulting in silent mutations: C894T in exon 5, C1134T in exon 7, and C1476T in exon 9. These do not alter the structure or the expression of the protein. It is unlikely that mutations in the coding region and splice junction of GLRA2 gene are associated with schizophrenia and other psychiatric diseases. PMID- 11409701 TI - The -141C Ins/Del polymorphism of the dopamine D2 receptor gene is not associated with either migraine or Parkinson's disease. AB - Abnormalities in dopamine neurotransmission at the dopamine D2 receptor (DRD2) have been implicated in both migraine and Parkinson's disease. Positive associations have also been found between polymorphisms within the DRD2 gene and both of these conditions. The -141C Ins/Del polymorphism in the DRD2 receptor gene is a putative functional polymorphism. The purpose of this study was to determine whether it and any genes in linkage disequilibrium with this marker are involved in either of these conditions. We have compared the genotype and allele frequencies of the -141C Ins/Del polymorphism in 200 migraineurs and 260 Parkinson's disease cases with 464 controls. We have found no association between the receptor gene and either condition (P = 0.89 and P = 0.56 respectively). Our findings do not support the hypothesis that this polymorphism is involved in the aetiology of migraine or Parkinson's disease. PMID- 11409702 TI - Exploring the complex relationship between adolescent sexual offending and sex chromosome abnormality. AB - The nature and importance of the relationship between sex chromosome abnormalities (SCAs) and sexual maladaptive behaviour is uncertain. When considering the aetiology of sexual offending behaviour, the importance of sex chromosome disorder lies in its biopsychological manifestations and in its complex interactions with external influences. At the Adolescent Forensic Service, in keeping with previous research in institutional settings, we found a higher pick-up rate of SCAs among sexual offenders (5/121) than would be expected in an unbiased community sample (1.2/1000 male livebirths; Jacobs et al. (1992)). We present descriptive data on five patients with SCAs out of a total of 121 sexual offenders who presented to the Adolescent Forensic Service over a 6-year period. We discuss the biopsychosocial features of these five patients and compare them with the remainder of the sexual offenders in the series. We discuss the advantages of early diagnosis and the need for professional vigilance by adolescent forensic psychiatrists, child and adolescent psychiatrists, paediatricians and clinical geneticists. PMID- 11409703 TI - No leucine(7)-to-proline(7) polymorphism in the signal peptide of neuropeptide Y in Japanese population or Japanese with alcoholism. AB - We have screened 200 Japanese workers and 105 Japanese patients with alcoholism for the mutation in the signal peptide of pre-pro-neuropeptide Y resulting in a substitution of proline for leucine at position 7. This polymorphism was reported in the Finnish and Dutch populations recently. None of our subjects displayed the mutation at this site. Therefore, this allele does not play any role in the development of alcoholism in the Japanese population. PMID- 11409704 TI - LDL cholesterol angiotensin II interactions in atherosclerosis. PMID- 11409705 TI - JMM, past and present. Meningitis cerebro-spinalis epidemica. PMID- 11409706 TI - Cubilin, a multifunctional epithelial receptor: an overview. AB - Cubilin is a 460-kDa endocytic receptor coexpressed with megalin, a multiligand receptor of the low-density lipoprotein receptor gene family, at the apical pole of epithelial cells in the renal proximal convoluted tubule, visceral yolk sac, ileum, and placenta. The structure of cubilin is unique: it lacks a transmembrane domain and requires megalin for its internalization. The accumulation of 27 interactive CUB domains provides the potential for multiple, possibly independent interactions and functions. Cubilin is involved in the intestinal absorption of vitamin B12, the catabolism of apolipoprotein A-I by the proximal convoluted tubule and more generally in renal protein reabsorption. The role of cubilin on fetomaternal interfaces is not defined but may be related to its ability to bind and internalize high density lipoproteins. PMID- 11409707 TI - Limitin: an interferon-like cytokine without myeloerythroid suppressive properties. AB - Limitin is an interferon (IFN)-like cytokine that we recently identified and cloned on the basis of its ability to arrest the growth or kill lympho hematopoietic cells. This 182 amino acid protein has approximately 30% sequence identity with IFN-alpha, IFN-beta, and IFN-omega. Limitin binds to the IFN alpha/beta receptors and induces IFN regulatory factor-1, thereby indicating that limitin constitutes a new prototype of the type I IFN family. As with previously known IFNs, limitin inhibited B lymphopoiesis in vivo as well as in vitro. In addition, limitin not only modified the proliferation and function of peripheral T lymphocytes, natural killer cells, and bone marrow stromal cells but also had antiviral activity. Therefore, limitin is a multifunctional cytokine with several potential cellular targets. Because to date we have found no influence of limitin on normal myeloid and erythroid progenitors, limitin is unique among the IFNs. Type I IFN family contains IFN-alpha, IFN-beta, IFN-omega, and IFN-tau, and IFN alpha is composed of at least 14 subtypes. All IFNs have anti-proliferative, immunomodulatory, and antiviral effects and influence to each other in the body. Limitin should play a role in the complex IFN network, and its human homologue would be useful as a therapeutic agent if it lacked myelosuppressive activity. PMID- 11409708 TI - Blood pressure response to angiotensin II, low-density lipoprotein cholesterol and polymorphisms of the angiotensin II type 1 receptor gene in hypertensive sibling pairs. AB - Blood pressure (BP) response to infused angiotensin II (Ang II) has been widely used to characterize hypertensive subjects. High cholesterol levels have recently been found to enhance this response in young men, suggesting an important new link between atherosclerosis and hypertension. The present study assessed the familial resemblance of the BP response following an Ang II infusion and measured the factors affecting the trait in a large set of hypertensive men and women. After a low-salt diet for 7 days a 30-min infusion of Ang II was administered to 218 white hypertensive patients (28 singletons, 80 sibling pairs, 10 trios). Age and gender were significantly correlated to the Ang II systolic but not to the diastolic BP response. Conversely, cholesterol level and especially low-density lipoprotein (LDL) were correlated to both systolic and diastolic changes. Multivariate analysis showed that age, gender, and LDL were the three parameters that explained the systolic BP change whereas plasma LDL remained the only variable significantly correlated to the diastolic BP change. Significant familial resemblances in the Ang II induced systolic and diastolic BP response were observed, especially in female pairs. On this limited number of subjects, suggestive evidence for association and linkage was found between the trait, A1166C, and (CA)n repeat polymorphisms of the Ang II type 1 receptor (AT1R) gene. In conclusion, the Ang II induced BP change is strongly related to plasma LDL in hypertensive men and women, stressing the importance of the lipid profile as a contributor to BP regulation. Familial resemblance of this intermediate phenotype is sex dependent and may be partly explained by polymorphisms of the AT1R gene. PMID- 11409709 TI - Efficacy of cationic liposome-mediated gene transfer to mesangial cells in vitro and in vivo. AB - Mesangial cells represent a major target for gene transfer approaches to the kidney. To establish a liposome-based system for transfection of mesangial cells we analyzed the efficacy and toxicity of different cationic liposomes and other nonviral transfection methods in primary cultures of rat and human mesangial cells using the Escherichia coli beta-galactosidase (lacZ) gene as a marker. In addition, an expression vector containing a human renin cDNA under the control of the cytomegalovirus immediate-early promoter/enhancer was generated, introduced into mesangial cells, and assayed in a system of transient gene expression. In vivo, gene transfer was studied after infusion of liposome/DNA complexes in the kidney of rats via the renal artery. Transfection efficiency ranged from 5.5% with DMRIE Liposomes in rat mesangial cells to 1.1% with LipofectAmine liposomes in human mesangial cells. Cytotoxicity following transfection was dependent on the transfection method. Transfection with the human renin expression vector led to the secretion of 11 pg/10(4) cells/48 h human renin in rat mesangial cells, 3,600 pg/10(4) cells/48 h in 293 cells, and 113 pg/10(4) cells/48 h human renin in opossum kidney cells. In vivo, infusion of liposomes was accompanied by nephrotoxicity and did not result in marker gene expression. Together the data demonstrate that cationic liposomes are useful tools for transferring genes into mesangial cells, including human mesangial cells. Cationic liposomes provide a functional system for the synthesis and secretion of human renin in mesangial cells and other mammalian kidney cells. The current limitation of the evaluated liposomes for an efficient in vivo gene transfer to mesangial cells is the toxicity upon intrarenal arterial administration. PMID- 11409710 TI - Disease resistant, NOD-related strains reveal checkpoints of immunoregulation in the pancreas. AB - The autoimmune diabetic NOD mouse serves as a model for human type 1 diabetes. Disease development is due to islet beta cell destruction in the context of immune cell infiltration of islets and inflammatory changes throughout the pancreas. In the present study we tried to identify immune reactivity patterns in the pancreas associated with diabetes resistance in NOD-related mouse strains. The pancreata of diabetes-prone female NOD/LtJ, NOD/Bom and of genetically related but diabetes-resistant strains; NOR, NON, NON.NOD-H2g7, NOD.NON-H-2nbl were obtained at the age of 70 days for semiquantitative analysis of insulitis and of mRNA expression by reverse transcriptase PCR. In addition, the response to a single dose of cyclophosphamide for synchronizing and accelerating the progression of insulitis was determined. The progression of insulitis and immune gene expression in response to cyclophosphamide revealed characteristic differences between the six strains. NOD/LtJ and NOD/Bom mice were found significantly to upregulate pancreatic IL-12p40 and IL-18 expression after cyclophosphamide treatment, followed by an increase in IFN-gamma mRNA levels. In contrast, the two MHC-haplotype H-2nbl expressing strains either up-regulated neither IL-12/IL-18 nor IFN-gamma gene expression. The two strains sharing MHC haplotype H-2g7 expression with NOD did respond to cyclophosphamide with IL 12p40/IL-18 gene expression. However, NON.NOD-H-2g7 mice failed to progress to IFN-gamma gene expression. NOR mice progressed to IFN-gamma expression but exhibited sustained IL-4 gene expression. Only severe intra-insulitis was associated with the expression of inducible NO synthase. The comparison of diabetes-prone and diabetes-resistant strains revealed three checkpoints of immune regulation in the pancreas. The earliest checkpoint is the induction of an IL-12p40/IL-18 response in innate immune or antigen-presenting cells. The next level of control is at the induction of IFN-gamma gene expression, and a third checkpoint is the maintenance or loss of antagonistic Th2 type reactions. PMID- 11409711 TI - A polymorphism, L162V, in the peroxisome proliferator-activated receptor alpha (PPARalpha) gene is associated with lower body mass index in patients with non insulin-dependent diabetes mellitus. AB - This study examined the effect a polymorphism (L162V) in the gene for peroxisome proliferator activated receptor (PPAR) alpha in the development of non-insulin dependent diabetes mellitus (type 2 DM), obesity and hyperlipidaemia. The frequency of the L162V polymorphism in the PPARalpha gene was determined in 370 morbidly obese patients who underwent gastric banding surgery, 154 patients attending a type 2 DM clinic, 188 patients attending a lipid clinic and 199 healthy blood donors. The overall frequency of the V allele of the L162V polymorphism was 0.06. There were no significant differences in the allele frequency between patients with morbid obesity, hyperlipidaemia, type 2 DM and healthy controls, suggesting that it does not play a major role in the development of these conditions. The polymorphism was associated with a lower body mass index (BMI) in two independently recruited groups of patients with type 2 DM. There was no effect of the polymorphism on subjects without type 2 DM. Thus a polymorphism in PPARalpha protects type 2 DM patients from the overweight which is frequently associated with their condition. PMID- 11409712 TI - Asialofetuin liposome-mediated human alpha1-antitrypsin gene transfer in vivo results in stationary long-term gene expression. AB - The development of nonviral vectors for in vivo gene delivery to hepatocytes is an interesting topic in view of their safety and tremendous gene therapy potential. Since cationic liposomes and liposome uptake by receptor-mediated mechanisms could offer advantages in the efficacy of liposome-mediated gene transfer, we studied the effect of liposome charge (anionic vs. cationic) and the covalently coupled asialofetuin ligand on the liposome surface in mediating human alpha1-antitrypsin (hAAT) gene transfer to mice in vivo. The changes in liposome charge were made by adding the following lipids to the backbone liposomes: anionic phosphatidylserine, cationic N-[1-(2,3-dioleoyloxy)propyl]-N,N,N trimethyl-ammonium methylsulfate or a lipopeptide synthesized from dipalmitoylphosphatidylethanolamine and covalently coupled to the cationic nuclear localization signal peptide. Two plasmids containing the hAAT gene were used: pTG7101, containing the complete genomic sequence of the human gene driven by the natural promoter, and p216, containing the human hAAT cDNA under the control of the CMV promoter. The results indicate that both untargeted anionic and cationic liposomes mediate plasma levels of hAAT that decline over time. However, asialofetuin liposomes increase the plasma levels of hAAT and can mediate long-term gene expression (>12 months) with stationary plasma levels of protein. Results from quantitative and qualitative reverse transcriptase polymerase chain reaction match those from protein plasma levels and confirm both the human origin of the message and the liver as source of the protein. The use of asialofetuin liposomes in hepatic gene therapy may both increase and prolong in vivo gene expression of hAAT and other clinically important genes. PMID- 11409713 TI - T cell responses to type 1 diabetes related peptides sharing homologous regions. AB - Glutamic acid decarboxylase (GAD) 65 is a major autoantigen in type 1 diabetes. Regions of homology exist between GAD65 (residues 250-273) and the Coxsackie P2-C protein (residues 28-50) and between GAD65 (residues 506-518) and proinsulin (residues 24-36), and each of these has been reported to be a diabetes-associated T cell target. The aim of this study was to determine whether the homologous regions are shared targets of T lymphocyte reactivity in individual patients with type 1 diabetes. T cell proliferation against the corresponding peptide pairs, GAD254-276 and Coxsackie P2-C32-54 and GAD506-518 and proinsulin24-36, were measured in peripheral blood mononuclear cells from 26 patients with newly diagnosed type 1 diabetes and 24 control subjects. Responses with stimulation indices higher than 3 were found against each of the antigens tested in both patients and control subjects, and no differences were observed between groups. A strong positive correlation was found between responses to the corresponding peptide pairs GAD254-276 and Coxsackie P2-C32-54 (r=0.77, P<0.0001), and between responses to the corresponding peptide pairs GAD506-518 and proinsulin24-36 (r=0.66, P<0.0001). However, a similar correlation was also observed between responses to the noncorresponding pairs Coxsackie P2-C32-54 and proinsulin24-36 (r=0.82, P<0.0001), Coxsackie P2-C32-54 and GAD506-518 (r=0.82, P<0.0001), and GAD254-276 and proinsulin24-36 (r=0.83, P<0.0001). Strikingly, increased responses to peptides were found almost exclusively in subjects with high stimulation indices against the recall antigen tetanus toxoid, further suggesting that peripheral blood T cell responses are related to a general subject hyperreactivity. These data suggest that proliferative T cell responses to peptides containing putative autoreactive epitopes of GAD65 and proinsulin are not specific for type 1 diabetes, that correlation between T cell reactivity to peptides is not restricted to those containing homologous regions, and that non antigen-specific factors are important determinants of in vitro measurements of T cell reactivity. PMID- 11409714 TI - Effect of ketamine on spinal cord nociceptive transmission in normal and monoarthritic rats. AB - The effects of systemically and intrathecally administered ketamine on spinal wind-up of normal and monoarthritic rats were studied by using C-fiber reflex responses evoked by repetitive (0.6 Hz) electric stimulation. Both systemic and intrathecal ketamine induced dose-dependent depression of wind-up activity in normal rats, as revealed by the dose-related inhibitory effects of the drug. At the same intraperitoneal doses, ketamine produced a greater inhibitory effect on wind-up activity of monoarthritic rats, compared to normal animals. The intrathecal administration of ketamine also produced wind-up inhibition, the efficacy being higher in the monoarthritic rats. Results indicate that ketamine depresses spinal wind-up, specially in rats submitted to chronic pain, probably due to its antagonistic properties on dorsal horn NMDA receptors, which play a crucial role in the maintenance of chronic pain. PMID- 11409715 TI - Effect of auditory cortex lesions on NADPH-diaphorase staining in the inferior colliculus of rat. AB - Projections from the auditory cortex (AC) in the rat terminate in the dorsal cortex (DC) and in the external cortex (EC) of the inferior colliculus (IC), areas which exhibit a moderate number of nicotinamide-adenine dinucleotide phosphate-diaphorase (NADPH-d) positive neurons. NADPH-d co-localizes with nitric oxide synthase, which is responsible for the production of the transcellular messenger, nitric oxide. Changes in NADPH-d staining in the IC were found after unilateral lesions of the AC. Lesions resulted in a reduction in NADPH-d staining in neurons and neuropil within the ipsilateral DC and EC with the maximum reduction occurring 3-4 days after lesion. The reduction in NADPH-d staining in the contralateral IC was less pronounced. Lesions affecting auditory areas Te 1 and Te 3 produced the largest decrease in NADPH-d staining in neurons and neuropil. This finding may be related to the abolition of the influence of glutamatergic corticocollicular and commissural pathways. PMID- 11409716 TI - Human primary auditory cortex in women and men. AB - Specific patterns of anatomical symmetry or asymmetry have been associated with sex differences in human brain structure and function. An observer-independent cytoarchitectonic method for the quantification of cell volume densities and areal borders was used to investigate the size and microstructure of primary auditory cortex (Brodmann area 41) in female (n = 14) and male (n = 13) postmortem brains. The total brain volume-adjusted volume of the primary auditory cortex was significantly larger in women than in men bilaterally. Inverse asymmetry towards the right side, as opposed to well-known asymmetries towards the left side, was more frequent in women. Laminar cell volume densities of BA 41 showed no gender effect. The morphometric data confirm (in part) gender differences in the cerebral organization of primary auditory cortex. PMID- 11409717 TI - Test-retest reliability of DSC MRI CBV mapping in healthy volunteers. AB - Dynamic susceptibility contrast magnetic resonance imaging (DSC MRI) can be used to generate high resolution maps of cerebral blood volume (CBV). To determine the test-retest reliability, CBV was measured in eight volunteers on two occasions, separated by 4 weeks. The mean ratio (scan 2/scan 1) for 72 cortical regions of interest (ROIs) was 1.03, with a coefficient of variation of 14%. The correlation between the first and second scans was 0.73 (p < 0.0001; 95%). In five hand-drawn ROIs, the mean ratio was 1.08, with a coefficient of variation of 12%. The correlation between scans was 0.84 (p < 0.0001). The data presented here support the hypothesis that DSC MRI CBV mapping has acceptable test-retest reliability. PMID- 11409718 TI - Oscillation of pain intensity during adenosine infusion. Relationship to beta endorphin and sympathetic tone. AB - Adenosine is a neuromodulator with both excitatory and inhibitory effects dependent in part upon preconditions; it can act as an algesic or an analgesic agent. Previously we found variations of pain intensity during constant infusion of adenosine. We therefore quantified pain intensity during constant infusion of adenosine at a rate of 140 microg/kg/min intravenously in healthy volunteers, placebo controlled, double blind, and the relation to hemodynamic, vasomotor and sudomotor responses of the sympathetic nervous system and to the role of peripheral beta-endorphin response. The perceived chest pain during adenosine infusion showed an oscillatory pattern. Painful periods of about 30s were interrupted by painfree periods, and pain was always preceded by an increase in vasomotor sympathetic activity and by increased sudomotor activity. Plasma beta endorphin values were heterogenous but exhibited an increase during infusion. PMID- 11409719 TI - Opposite changes in adenosine A1 and A2A receptor mRNA in the rat following sleep deprivation. AB - Extracellular levels of adenosine increase in basal forebrain following prolonged wakefulness. Moreover, perfusion of adenosine into basal forebrain increases sleep. In this study we have examined the adenosine receptor subtypes, A1 and A2A, for changes in the levels of mRNA using RT-PCR and in situ hybridization and the receptor ligand binding efficiency using autoradiography following 3 and 6 h of sleep deprivation. We observed that A1 receptor mRNA levels increased in basal forebrain with no changes in other forebrain areas examined. A1 receptor binding was not affected. A2A receptor mRNA and ligand binding were undetectable in basal forebrain. However, in the olfactory tubercle, A2A mRNA and receptor binding decreased significantly. Based on the significant increase in the A1 but not in A2A receptor, we hypothesize that the effects of sleep deprivation-induced increased adenosine are mediated by A1 receptor in basal forebrain of rats. PMID- 11409720 TI - A functional MRI study of face recognition in patients with prosopagnosia. AB - An fMRI investigation was conducted to determine whether patients with impaired face recognition, a deficit known as prosopagnosia, would show functional activation in the fusiform gyrus, the neural substrate for face processing, when viewing faces. While the patients did show activation in the fusiform gyrus, with significantly more voxels in posterior areas than their control subjects, this activation was not sufficient for face processing. In one of the patients, the posterior activation was particularly evident in the left hemisphere, which is thought to be involved in feature-based strategies of face perception. We conclude that an increased reliance on feature-based processing in prosopagnosia leads to a recruitment of neurons in posterior regions of the fusiform gyrus, regions that are not ideally suited for processing faces. PMID- 11409721 TI - Co-localization of 5-HT(1B/1D/1F) receptors and glutamate in trigeminal ganglia in rats. AB - Anti-migraine triptan drugs are 5-HT(1B/1D) receptor agonists which are thought to block the neurotransmitter/neuropeptide release from sensory nerve terminals and directly constrict blood vessel smooth muscles. In the present study, we have investigated the anatomical basis for a possible modulation of glutamate release from trigeminal ganglion neurons by 5-HT(1B/1D) receptor agonists and by 5-HT1F receptor agonists, using double immunohistochemical staining technique in the rat. The majority of 5-HT1B, 5-HT1D or 5-HT1F receptor positive neurons were also glutamate positive, but both 5-HT1B, 5-HT1D or 5-HT1F receptor single-labeled and glutamate single-labeled neurons were observed. These results suggest that 5 HT(1B/1D/1F) receptor agonists may modulate glutamate release, and that one mechanism of their anti-migraine action could be the blockade of glutamate release. PMID- 11409722 TI - Efficacy of background GABA uptake in rat hippocampal slices. AB - GABA uptake is crucial for the termination of inhibitory synaptic events. In addition, GABA transporters may also control the level of diffusely distributed GABA in the extracellular space. We analysed this function by superfusing rat hippocampal slices with different concentrations of GABA. Whole-cell patch clamp recordings of CA1 pyramidal cells revealed small increases in chloride conductance at 5-10 microM GABA which increased dramatically upon addition of the GABA uptake blocker tiagabine. Tiagabine alone induced a significant chloride conductance indicating that spontaneous release of GABA in hippocampal slices is neutralized by GAT-1, the main hippocampal GABA transporter. Thus, GAT-1 clears the extracellular space in the hippocampus from diffusely distributed GABA with high efficacy. PMID- 11409723 TI - VRL-1 immunoreactivity in the rat cranial autonomic ganglia. AB - Immunohistochemistry for VRL-1, a newly cloned capsaicin-receptor homologue, was performed on the rat cranial autonomic ganglia. The immunoreactivity (ir) was detected in the majority of neurones in the pterygopalatine (66%) and submandibular ganglia (68%). In the tongue and carotid body, parasympathetic neurones contained VRL-I ir. In the superior cervical ganglion, only 2% of postganglionic sympathetic neurones showed the immunoreactivity. VRL-1-ir nerve endings could not be detected in their peripheral tissues. These findings may suggest that VRL-1 has functions within neuronal cell bodies of the cranial autonomic ganglia. PMID- 11409724 TI - NO synthase inhibitors reduce opioid desensitization in rat locus coeruleus neurons in vitro. AB - The aim of this study was to examine by electrophysiological techniques whether nitric oxide (NO) is involved in the development of desensitization to the opioid agonist Met5-enkephalin (ME) in locus coeruleus neurons from rat brain slices. Bath perfusion with ME (0.05-1.6 microM) caused a concentration-dependent reduction in the firing rate of locus coeruleus cells, whereas perfusion with a high concentration of ME (10 microM) desensitized the inhibitory effect of subsequent ME (0.8 microM) applications. However, in slices perfused with the NO synthase inhibitors 7-NI (100 microM), L-NAME (100 microM) or L-NA (100 microM) the ME (10 microM)-induced opioid desensitization was strongly attenuated. The effect of L-NAME was prevented by administration of L-arginine (100 microM). These results suggest that nitric oxide may contribute to opioid desensitization in locus coeruleus neurons. PMID- 11409725 TI - Selective up-regulation of 5-HT(1B/1D) receptors during organ culture of cerebral arteries. AB - 5-Hydroxytryptamine (5-HT) is thought to be involved in migraine headache and the pathophysiology of cerebrovascular diseases. Previous data show that organ culture induces a phenotypic change in cerebral vessels. Therefore we investigated if these changes also applied for the vasoconstrictive 5-HT receptors. Rat cerebral arteries express 5-HT2 receptors. Using organ culture we observed a phenotypic change with a selective up-regulation of 5-HT(1B/1D) receptors. This was revealed by an increased sensitivity to the selective 5 HT(1B/1D) agonist 5-CT after organ culture (pEC50(fresh) 5.6+/-0.2 and pEC50(cultured) 6.8+/-0.4). The response was inhibited by the 5-HT(1B/1D) selective antagonist GR55562 (pEC50(fresh) 5.1+/-0.2 and pEC50(cultured) 6.0+/ 0.3). The organ model might mimic the phenotypic changes during cerebrovascular diseases. PMID- 11409726 TI - Cortical map asymmetries in the context of transcallosal excitatory influences. AB - There is long-standing disagreement among experimentalists about whether transcallosal interhemispheric influences are primarily excitatory or inhibitory. Past computational models exploring this issue have encountered a similar dilemma: inhibitory callosal influences best explain hemispheric functional asymmetries, but excitatory callosal influences best explain transcallosal diaschisis. We recently hypothesized that this dilemma might be resolved by assuming excitatory callosal influences and a subcortical mechanism for cross midline inhibition. Here we explore the feasibility of this hypothesis by examining a model of map formation in corresponding left and right cortical regions. The results show for the first time that both map asymmetries and diaschisis-like changes can be produced in a single model, suggesting that subcortical inhibitory processes may contribute more to asymmetric cortical functionality than is generally recognized. PMID- 11409727 TI - Expression of NeuroD and TrkB in developing and aged mouse olfactory epithelium. AB - To better understand the roles of NeuroD, a member of the basic helix-loop-helix transcription factor family, during the differentiation of olfactory receptor neurons, we studied the expression of NeuroD in developing and aging mouse olfactory epithelium (OE). During embryonic period, NeuroD expression is confined in the basal compartment of OE. During neonatal period, NeuroD expression is detected in the middle compartment and in the basal compartment of OE. In the adult, the number of NeuroD expressing cells in the basal compartment significantly decreased, while the NeuroD-positive cells in the middle compartment was maintained throughout lifetime. This dual phase expression pattern of NeuroD suggests multiple roles of NeuroD in the neurogenesis of ORNs. PMID- 11409728 TI - Cholecystokinin-8 promotes recovery of sympathectomy induced by 6-hydroxydopamine in adult mice. AB - We used an experimental model of sympathetic neuropathy to investigate the effects of intraperitoneal cholecystokinin-8 (CCK-8) administration on the recovery of injured peripheral neurones. After treatment of adult mice with 6 hydroxydopamine (6-OHDA), which known to induce peripheral sympathectomy, nerve growth factor (NGF) in peripheral tissue first increased and then rapidly decreased to baseline levels. Following this observation, sympathectomised mice were treated with CCK-8 starting when the NGF levels lowered toward the control value. Our results show that injections with 8 nmol/kg of CCK-8 promote not only recovery of noradrenergic innervation but also NGF and neuropeptide Y (NPY) synthesis in peripheral tissue. This latter observation suggests that the effect of CCK-8 might be mediated through the stimulation of NGF synthesis. PMID- 11409729 TI - Neuroprotective effects of mild hyperthermia prior to focal ischemia in conscious rats. AB - Hyperthermia during or after stroke is known to worsen neuronal damage. Paradoxically, when hyperthermia precedes stroke, it can protect against a subsequent ischemic insult. Other stressors including restraint also have a similar pre-conditioning effect. In the present study, we report the unanticipated finding that conscious rats, restrained for the purpose of intravenous infusion, had markedly reduced neuronal and functional deficits after middle cerebral artery occlusion compared with unrestrained rats. Restrained rats had significantly higher body temperature prior to stroke than unrestrained rats. The findings suggest restraint leading to mild hyperthermia may be sufficient to induce adaptive processes which protect against subsequent ischemia. PMID- 11409730 TI - Dispositional pessimism and amygdala activity: a PET study in healthy volunteers. AB - The present study used scores from Seligman's Attribution Style Questionnaire and [15O] water positron emission tomographic measurements of regional cerebral blood flow (rCBF) to investigate the relation between individual differences in dispositional pessimism and amygdala activity. During scanning 13 healthy non snake-phobic females passively viewed a snake videotape. Using one-tailed tests, significant negative correlations were evident between pessimism scores, with low scores reflecting relatively more pessimism, and right (r=-0.60; p=0.014) and left amygdala rCBF (r=-0.53; p=0.032). These results extend previous neuroimaging findings in healthy subjects indicating a role for the amygdala in transient negative emotional states, and suggest that this multimodal brain region also is involved in more durable negative affects such as dispositional pessimism. PMID- 11409731 TI - Activation of phosphatidylinositol 3 kinase by muscarinic receptors in astrocytoma cells. AB - Stimulation of Gq-coupled acetylcholine muscarinic receptors leads to proliferation of astroglial cells, but the signal transduction pathway(s) that mediate this mitogenic response have not been fully elucidated. In this study, we report on the ability of carbachol to stimulate the phosphorylation of Akt/PKB, an important target of phosphatidylinositol 3 kinase (PI3 kinase) in 1321N1 human astrocytoma cells. Carbachol induced a dose-dependent phosphorylation of Ser473 on Akt, peaking after 15 min. This effect was mediated by activation of the M3 subtype of muscarinic receptors and was inhibited by two PI3 kinase inhibitors. Inhibitors of protein kinase C, mitogen-activated protein kinase and p70S6 kinase, had no effect on carbachol-induced Akt phosphorylation. Carbachol-induced DNA synthesis was strongly inhibited by two PI3 kinase inhibitors, wortmannin and LY294002, suggesting that PI3 kinase activation plays an important role in carbachol-induced proliferation 1321N1 astrocytoma cells. PMID- 11409732 TI - Normal apoptosis levels in mice expressing one alpha7 nicotinic receptor null and one L250T mutant allele. AB - The alpha7 nicotinic receptor (nAChR) is a ligand-gated ion channel mediating cholinergic transmission throughout the nervous system. To further characterize the function of this receptor, we generated mice expressing the alpha7 L250T nAChR mutation and demonstrated that homozygous (T/T) L250T mice die within 24 h of birth and display extensive apoptosis and abnormal layering within their cortex. We now demonstrate that mice with one alpha7 null and one L250T allele ( /T) show little apoptosis and normal development of their cortex yet exhibit the same lethal phenotype as T/T mice. Furthermore, L250T mice show normal levels of apoptosis in other nervous system regions expressing alpha7 nAChRs. These results suggest that apoptosis is not the cause of death for L250T neonatal mice. PMID- 11409733 TI - Preserved stimulus deviance detection in Alzheimer's disease. AB - Aging attenuates automatic auditory discrimination to duration change, whereas frequency change detection is relatively unimpaired in aging and in Alzheimer's disease (AD). Here we studied with a whole-head magnetometer whether cortical auditory discrimination to duration change as shown by magnetic mismatch negativity (MMNm) response is impaired in AD. Twenty AD patients with mild to moderate cognitive impairment and 18 age-matched healthy subjects were monaurally presented a sequence of frequent standard tones embedded with occasional deviants with shorter duration. MMNm was significantly delayed in the left hemisphere ipsilaterally to the ear stimulated in the patient group, whereas the MMNm amplitudes over both hemispheres were quite similar in both groups. This suggests that although MMNm is delayed in the left hemisphere, the automatic discrimination to duration change in the auditory cortex is not attenuated in the early stages of AD. PMID- 11409734 TI - UVB irradiation-induced apoptosis increased in lymphocytes of Huntington's disease patients. AB - Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by CAG repeat expansion in the IT-15 gene coding for huntingtin. The mechanism of neuronal degeneration induced by the mutant huntingtin is not known. Apoptosis may play a role in it. Huntingtin is widely expressed in the cells, so abnormalities can be expected also in non-neural tissue. We examined the susceptibility of lymphocytes from HD patients, asymptomatic carriers and normal individuals to UVB irradiation-induced apoptosis. Lymphocytes from eight HD patients and two asymptomatic carriers showed increased apoptotic cell death compared to controls. Our results suggests that sensitivity of HD cells to induced apoptosis is not restricted to neurons. PMID- 11409735 TI - Analysis of pain-related somatosensory evoked magnetic fields using the MUSIC (multiple signal classification) algorithm for magnetoencephalography. AB - We evaluated the effectiveness of the Multiple Signal Classification (MUSIC) algorithm by analysing pain-related somatosensory-evoked magnetic fields (SEFs) by 148-channel whole-head-type magnetoencephalography. MUSIC peaks of middle latency components were located around the primary somatosensory cortex (SI), contralateral to the stimulated finger. Long latency components were located around the bilateral secondary somatosensory cortices (SII) and cingulate gyri. Peaks at the SII and cingulate gyri were more prominent on very painful and moderately painful stimulation than on weak stimulation. The results were in very good agreement with results from single dipole estimation. These findings suggest that the MUSIC algorithm could be a useful tool for analysis of pain-related SEFs. PMID- 11409736 TI - Cerebral protection by hypoxic preconditioning in a murine model of focal ischemia-reperfusion. AB - Sublethal periods of hypoxia or ischemia can induce adaptive mechanisms to protect against subsequent lethal ischemic insults in a process known as ischemic preconditioning. In the present study, we developed a murine model of cerebral preconditioning using several common strains of adult mice. Animals were exposed to sublethal hypoxia (11% oxygen for 2 h) 48 h prior to a 90 min period of transient focal middle cerebral artery occlusion, induced by an intraluminal filament; injury was assessed 24 h later by TTC staining. Infarct volume in hypoxia-preconditioned animals was reduced 46%, 58%, and 64% in C57Bl/6, 129SvEv, and Swiss-Webster ND4 mice relative to their respective untreated controls. This non-invasive murine model of ischemic tolerance should be useful for elucidating the molecular basis of this protection using transgenic and knockout mice. PMID- 11409737 TI - Eyes first! Eye processing develops before face processing in children. AB - Faces and eyes are critical social stimuli which adults process with ease, but how this expertise develops is not yet understood. Neural changes associated with face and eye processing were investigated developmentally using ERPs (N170), in 128 subjects (4-15 year olds and adults). Stimuli included upright faces to assess configural processing, eyes and inverted faces to assess feature-based processing. N170 was present in the youngest children with similar patterns of face sensitivity seen in adults. Development of N170 to upright faces continued until adulthood, suggesting slow maturation of configural processing. In contrast, N170 was shorter latency and much larger to eyes than faces in children and was mature by 11 years, suggesting the early presence of an eye detector, with a rapid maturational course. PMID- 11409738 TI - Genetic regulation of regional microstructure of the corpus callosum in late life. AB - In order to identify brain structural phenotypes that remain under significant genetic control in late adulthood, we examined the heritability of corpus callosum macrostructure (i.e. size) using MRI and microstructure (e.g. myelin) using diffusion tensor imaging in 15 monozygotic and 18 dizygotic twin pairs of elderly men. The relative proportion of genetic to environmental influences varied considerably by region and structural type and was 5:1 for callosal macrostructure, 3:1 for splenium microstructure, and 1:1 for genu microstructure. This is the first in vivo identification of quantifiable phenotypes of brain white matter microstructure and demonstrates significant and differential genetic regulation in old age, with anterior interhemispheric connecting pathways more susceptible than posterior pathways to environmental influences. PMID- 11409739 TI - Contralaterality of cortical auditory processing at the level of the M50/M100 complex and the mismatch field: a whole-head magnetoencephalography study. AB - Humans show a stronger cortical representation of auditory input at the opposite hemisphere each. To specify the temporal aspects of this contralaterality effect within the domain of speech stimuli, the present study recorded a series of evoked magnetic fields (M50, M100, mismatch field) subsequent to monaural application of stop consonant-vowel syllables using whole-head magnetoencephalography (MEG). The M50 components exhibited a skewed shape of cross-symmetrical distribution in terms of an initial maximum peak succeeded by a knot over the contralateral and a reversed pattern over the ipsilateral temporal lobe. Most presumably, this pattern of evoked fields reflects two distinct stages of central-auditory processing: (a) initial excitation of the larger contralateral and the smaller ipsilateral projection area of the stimulated ear; (b) subsequent transcallosal activation of the residual neurons, i.e. the targets of the non-stimulated ear, at either side. Previous studies using non-speech stimuli found contralaterality of central-auditory processing to extend to the M100 field. In contrast, a larger amplitude of ipsilateral M100 as compared to the respective opposite deflection emerged after stimulation of either ear. Finally, the computed magnetic analogues of mismatch negativity failed any significant laterality effects. These data provide first evidence for a distinct pattern of hemispheric differences at the level of the M50/M100 complex subsequent to monaural application of speech stimuli. PMID- 11409740 TI - Non-invasive single-trial monitoring of human movement-related brain activation based on DC-magnetoencephalography. AB - Neuroimaging techniques, such as fMRI, PET and near-infrared spectroscopy, monitor task-related neuronal activations in the brain indirectly through the associated neurovascular/metabolic responses. To assess the primary neuronal activations directly, magnetoencephalography was combined here with a mechanical modulation of the head-to-sensor position and signal separation via independent component analysis. In all of five subjects this approach allowed to monitor the time evolution of DC fields (<0.1 Hz) over the left hemisphere related to complex finger movements of the right hand alternating with rest periods (30 s each). Throughout the recording period of 30 min, stable task-related DC fields were recordable in a single-trial mode, i.e. without any averaging. DC-MEG opens up the possibility of analysing non-invasively cortical DC-activity also in stroke, migraine or epilepsy patients. PMID- 11409741 TI - Topography of orientation centre connections in the primary visual cortex of the cat. AB - The functional topography of lateral connections to orientation-centre zones was studied by optical imaging of intrinsic signals in combination with tracer injections (fluorescent beads and biocytin) and electrophysiological recordings. Three-dimensional reconstruction of anterogradely labelled axon terminals and retrogradely labelled somata revealed a uniform distribution across all orientations in a non-patchy manner. The overall lateral extent of the labelling was 3-4 mm in layer 3, that is about half of the extent observed for orientation domain connections in the same layer. These bulk injection data are in contrast with the reportedly sharp orientation tuning of neurons of centre zones and suggest that orientation specificity here does not require highly specific connections. Nonetheless, another plausible scenario is that orientation centre connections are orientation specific but their specificity present at the single cell level cannot be revealed by bulk labelling due to their large spatial overlap. PMID- 11409742 TI - Role of endogenous ATP at the incision area in a rat model of postoperative pain. AB - The aim of the present study is to characterize the role of endogenous ATP leaked from damaged cells in a rat model of postoperative pain using behavioural and immunocytochemical approaches. We found that systemic (i.v.) and local (incision area) administration of a P2 receptor antagonist, pyridoxalphosphate-6-azophenyl 2',4'-disulphonic acid (PPADS) before surgery significantly attenuated mechanical allodynia caused by an incision of the plantar surface of the hindpaw. Furthermore, PPADS significantly reduced the incision-evoked c-Fos protein expression, a marker of neuronal activity, in the dorsal horn of the spinal cord. The present findings suggest that excitatory signaling by endogenous ATP leaked from damaged cells via PPADS-sensitive P2 receptors is necessary for the induction of the postoperative pain characterized by mechanical allodynia. PMID- 11409743 TI - Effects of Gabapentin and Topiramate in primary rat astrocyte cultures. AB - We studied in vitro the effects of anticonvulsant drugs Gabapentin and Topiramate on the production of reactive oxygen species and nitric oxide (NO), the activity of glutamine synthetase (GS), and cell viability in primary cultures of rat cortical astrocytes which are intimately involved in the normal functioning of neurons. We investigated the effects of these drugs at concentrations within the therapeutic range (1, 10 and 50 microg/ml). We observed that, in cultured astrocytes, Gabapentin induced a weak increase in the biosynthesis of NO, a mild decrease in GS activity and cell viability, and minor induction of a stress condition. Topiramate was observed to induce even greater stressor effects on these cells. PMID- 11409744 TI - Adrenaline modulates multiple conductances in rat vagal motoneurones in vitro. AB - Whole cell recordings were undertaken from vagal motoneurones, including identified gastric vagal motoneurones, located within the medial regions of the dorsal vagal motonucleus of the rat medulla in vitro. The actions of adrenaline on individual channels expressed by these neurones were investigated. Adrenaline directly inhibits Ca2+ currents and delayed rectifier K+ currents and activates a sustained Na+ current. It also inhibits both Ca2+ activated non-selective cationic currents and Ca2+ activated K+ currents, the latter via inhibition of the underlying activating Ca2+ current. Since different sub-populations of vagal motoneurones express different complements of ion channels, this selective modulation of specific conductances by adrenaline may provide a mechanism by which adrenergic inputs, which project non-selectively throughout the DMV, could selectively control different aspects of vagal function. PMID- 11409745 TI - Glutamate-induced cobalt uptake reveals non-NMDA receptors in developing rat taste buds. AB - Non-NMDA type glutamate receptors are present in rat taste buds. However, the function of those receptors is not yet known. Developmental changes in the glutamate receptors in taste cells may provide clues to their functional role. We used a cobalt staining technique to determine at which stage in development functional non-NMDA glutamate receptors first appeared. Cobalt-stained taste bud cells first appeared in 20-day-old rats. The number of cobalt-stained cells increased with age and reached a maximum at 45 days. The shape of stained cells looked similar at all age groups. Cobalt-labeled cells appeared to be correlated with synaptic, not taste, glutamate receptors. PMID- 11409746 TI - Peripheral nerve regeneration through a long detergent-denatured muscle autografts in rabbits. AB - Muscle segments excised from rabbit biceps femoris muscles were treated with detergent sodium dodecyl sulphate to denature cellular constituents, and each was autografted in a 5 cm gap of the sciatic nerve in the same rabbit. Axonal regrowth through the grafts and reinnervation into the host sciatic nerves and muscles were studied morphologically, and electrophysiologically, 4 months after grafting. Regenerating axons accompanied by Schwann cells extended through basal lamina tubes of the grafts into the distal host nerves. Reinnervation of the tibialis anterior muscles by motor nerves was confirmed by recovery of the compound muscle action potentials (CMAP) and the reinnervation of the muscle spindles was demonstrated by electron microscopy. These findings indicated that the basal lamina tubes of denatured muscles were effective scaffolds through which the regenerating nerve fibers grew across as large a gap as 5 cm. PMID- 11409747 TI - Oxytocin receptors in non-human primate brain visualized with monoclonal antibody. AB - The identity of putative oxytocin receptors visualized in autoradiographic studies of primate brain is unclear because the ligand used is much less selective in primate than rodent brains. This study tests the feasibility of utilizing a new monoclonal antibody (MoAb) developed against human uterine OTRs to visualize OTRs in primate brain. A block containing ventral hypothalamus of cynomolgus macaque brain, paraformaldehyde, glutaraldehyde-fixed and paraffin embedded, and positive control tissue (human endometrium) were sectioned at 8 microm and studied with immunohistochemistry. OTRs were located in fibers in septal nucleus and in both cell bodies and fibers of preoptic area. These results indicate that OTRs in primate brain may be visualized with this MoAb, and are located in primate brain sites linked to the regulation of social behavior. PMID- 11409748 TI - Evidence of interactions between melanocortin and opioid systems in regulation of feeding. AB - The aim of our experiments was to study the presumed functional relationship between the melanocortin and opioid systems in the regulation of food intake. We determined that a non-selective opioid receptor antagonist, naltrexone, at relatively low doses, decreases food intake induced by i.c.v. agouti-related protein (Agrp). We also observed that peripheral injection of naltrexone at a dose known to produce anorexigenic responses induced c-Fos immunoreactivity in significantly more arcuate nucleus alpha-MSH neurons than observed in control animals. The results of our study support the notion that the melanocortin and opioid systems interact in the regulation of food intake. Based on these data we speculate that opioid peptides suppress alpha-MSH-dependent satiety mechanisms; conversely, it is possible that the orexigenic action of Agrp is mediated via opioid dependent circuitry. PMID- 11409749 TI - Different regulation of RGS2 mRNA by haloperidol and clozapine. AB - Expression of RGS2 mRNA was transiently up-regulated in rat striatum (25% in the medial part and 50% in the lateral part), in contrast to cingulate cortex and lateral septum, 30 min after acute treatment with haloperidol (2 mg/kg, i.p.). This effect disappeared 24 hours post-drug treatment, similar to the acute and strong up-regulation (700% at 30 min) of c-fos mRNA. RGS3, 5, 6, 8 or 9 mRNAs were not affected. Clozapine (20 mg/kg, i.p.) at an approximately equivalent dose of D2 receptor occupancy in the striatum did not significantly affect RGS and c fos mRNAs levels. We suggest that RGS2 mRNA expression may be differently up regulated in a region-specific manner by antipsychotics. PMID- 11409750 TI - Contributions of frontal and medial temporal regions to verbal episodic memory: a PET study. AB - Using PET, subtraction and correlation analysis were jointly employed to determine the specific and complementary contributions of frontal and medial temporal regions to verbal episodic encoding and retrieval processes. Subtraction analysis highlighted prefrontal rCBF increases which were predominantly left sided during intentional encoding and exclusively right-sided during retrieval, the latter being moreover associated with bilateral precuneus activation. However, significant correlation between rCBF values obtained during intentional encoding and performance scores obtained during retrieval concerned, among other regions, the left parahippocampal gyrus, which indicated that the higher the neuronal activity in this medial temporal region during encoding, the better the retrieval performance. PMID- 11409751 TI - The role of basal ganglia in reinforcement learning and imprinting in domestic chicks. AB - Effects of bilateral kainate lesions of telencephalic basal ganglia (lobus parolfactorius, LPO) were examined in domestic chicks. In the imprinting paradigm, where chicks learned to selectively approach a moving object without any explicitly associated reward, both the pre- and post-training lesions were without effects. On the other hand, in the water-reinforced pecking task, pre training lesions of LPO severely impaired immediate reinforcement as well as formation of the association memory. However, post-training LPO lesions did not cause amnesia, and chicks selectively pecked at the reinforced color. The LPO could thus be involved specifically in the evaluation of present rewards and the instantaneous reinforcement of pecking, but not in the execution of selective behavior based on a memorized color cue. PMID- 11409752 TI - Role of anterior temporal cortex in auditory sentence comprehension: an fMRI study. AB - Recent neuropsychological and functional imaging evidence has suggested a role for anterior temporal cortex in sentence-level comprehension. We explored this hypothesis using event-related fMRI. Subjects were scanned while they listened to either a sequence of environmental sounds describing an event or a corresponding sentence matched as closely as possible in meaning. Both types of stimuli required subjects to integrate auditory information over time to derive a similar meaning, but differ in the processing mechanisms leading to the integration of that information, with speech input requiring syntactic mechanisms and environmental sounds utilizing non-linguistic mechanisms. Consistent with recent claims, sentences produced greater activation than environmental sounds in anterior superior temporal lobe bilaterally. A similar speech > sound activation pattern was noted also in posterior superior temporal regions in the left. Envirornmental sounds produced greater activation than sentences in right inferior frontal gyrus. The results provide support for the view that anterior temporal cortex plays an important role in sentence-level comprehension. PMID- 11409753 TI - Elicitation of N400m in sentence comprehension due to lexical prosody incongruity. AB - The role of lexical prosody in the semantic integration of spoken sentences consisting of a quiz stem and an answer word was investigated analyzing the event related magnetic response, N400m. Three conditions regarding the relations between the quiz and the answer word were prepared: pitch-accent violation, phonemic violation and no violation. Both the pitch-accent and phonemic violations elicited significant N400m without any significant differences in the peak latency and magnitude of the equivalent current dipoles, suggesting that the role of pitch-accent in semantic integration is equivalent to that of phonemes. However, the rate of violation detection and the successful N400m source estimation were lower for the pitch-accent violation than for the phonemic violation, suggesting differential neural processes for the phonemic and prosodic cues. PMID- 11409754 TI - In vitro inhibitory effects of J-113397 on nociceptin/orphanin FQ-stimulated. AB - J-113397 (1-[(3R,4R)-1-cyclooctylmethyl-3-hydroxymethyl-4-piperidyl]-3-ethyl-1,3 dihydro-2H-benzimidazol-2-one) is a recently developed antagonist of the opioid receptor-like 1 (ORL1) receptor. We compared the in vitro functional profile J 113397 on [35S]guanosine 5'-O-(gamma-thio)triphosphate (GTPgammaS) binding to mouse brain with that of [Phe1psi(CH2-NH)Gly2]nociceptin(1-13)NH2 and naloxone benzoylhydrazone (NalBzoH). J-113397 antagonized nociceptin/orphanin FQ stimulated [35S]GTPgammaS binding to mouse brain with an IC50 value of 7.6 nM, but had no effect on basal [35S]GTPgammaS binding by itself. [Phe1psi(CH2 NH)Gly2]nociceptin(1-13)NH2 partially antagonized nociceptin/orphanin FQ stimulated [35S]GTPgammaS binding but showed agonistic activity on ORL1 by itself. NalBzoH showed antagonistic activity on ORL1 receptor but had significant agonistic activity on other opioid receptors at lower doses. Schild plot analysis demonstrated competitive antagonism of J-113397 on ORL1 receptor in mouse brain. A [35S]GTPgammaS binding study using ORL1 receptor-deficient mice confirmed the selective antagonism of J-113397 on ORL1 receptor. These data indicate that J 113397 is the most potent and selective antagonist of ORL1 receptor in mouse brain that has yet been reported, and therefore will be a useful tool for characterization of ORL1 receptors in the brain. PMID- 11409755 TI - The neural correlates of driving. AB - We studied 12 healthy subjects with fMRI while they performed a driving simulation task. In the active condition they steered the car themselves (driving), in the passive condition a person from outside the scanner was steering the car (passive driving). Common activations in both conditions were found in occipital and parietal regions bilaterally. Activity specifically associated with driving was found only in the sensorimotor cortex and the cerebellum. Compared to passive driving, activity during driving was reduced in numerous brain regions including MT/MST. It is concluded that simulated driving requires mainly perceptual-motor integration and that the limited cognitive capacity model of driving has to be revised. PMID- 11409756 TI - Topological analysis of striatal lesions induced by 3-nitropropionic acid in the Lewis rat. AB - 3-Nitropropionic acid (3NP) is a succinate dehydrogenase inhibitor classically used to create animal models of Huntington's disease (HD). However, the effects of this neurotoxin are highly variable in the Sprague-Dawley rat strain, impeding the interest of such model in neuroprotection assays. In the present study, we found that continuous s.c. infusion of 3NP in the Lewis rat strain produces homogeneous clinical impairments as well as highly reproducible striatal lesions according to their location and size. More especially, using quantitative reconstructions, we have determined, after 5 days of treatment, that the lesion was topologically reproducible in the lateral part of the striatum in all tested rats. Thus, 3NP-treated Lewis rat provides an improved animal model for testing neuroprotective strategies in HD. PMID- 11409757 TI - Abnormal P600 in heroin addicts with prolonged abstinence elicited during a working memory test. AB - The P600 component of event-related potentials, believed to be generated by anterior cingulate gyrus and basal ganglia, is considered as an index of aspects of second-pass parsing processes of information processing, having much in common with working memory (WM) systems. Moreover, dysfunction of these brain structures as well as WM deficits have been implicated in the pathophysiology of opioid addicts. The present study is focused on P600 elicited during a WM test in twenty heroin addicts with prolonged abstinence compared with an equal number of healthy controls. The results showed significantly prolonged latencies at right hemisphere, specifically at Fp2 abduction. Moreover, memory performance of patients did not differ from that of normal controls. These findings may indicate that abstinent heroin addicts manifest abnormal aspects of second-pass parsing processes as are reflected by the P600 latencies, elicited during a WM test. Additionally, the P600 might serve as a valuable investigative tool for a more comprehensive understanding of the neurobiological substrate of drug abuse. PMID- 11409758 TI - Galanin enhances noradrenaline-induced outward current on locus coeruleus noradrenergic neurons. AB - Interactions between galanin and noradrenaline (NA) were studied in a locus coeruleus (LC) slice preparation using intracellular recording technique. Both galanin (10-100 nM) and NA induced an outward current. Idazoxan, an alpha2 adrenoceptor antagonist, blocked the NA- but not the galanin-induced outward current, indicating that different receptors mediate these responses. Moreover, the outward current induced by NA was enhanced and prolonged by preincubation with a low concentration of galanin (0.05-0.1 nM), which had no detectable effect on the membrane current by itself. This sensitizing effect may be of physiological importance and could be caused by galanin released from dendrites and soma of galanin/NA neurons and/or from galanin afferents. Thus, besides a direct action of galanin on locus coeruleus neurons, our results also indicate an indirect, modulating effect. PMID- 11409759 TI - Common neural mechanism for processing onset-to-onset intervals and silent gaps in sound sequences. AB - Stimulus onset asynchrony (SOA) and inter-stimulus interval (ISI) are important factors in the perceptual organization of sound sequences. The present study tested whether these two temporal parameters are independently processed in the auditory system. Independence was studied by testing the additivity of mismatch negativity (MMN). Four conditions differing in their temporal regularities were administered: (1) constant SOA and ISI, (2) constant SOA and variable ISI, (3) constant ISI and variable SOA, and (4) variable SOA and ISI. The MMN elicited by simultaneous deviance from the constant SOA and ISI (Condition 1) was compared with an additive model calculated from the MMNs elicited in the other conditions. The amplitude of the MMN in Condition 1 was significantly larger than that of the modeled MMN, suggesting that SOA and ISI are processed by interactive or common neural mechanisms. PMID- 11409760 TI - Nocistatin, a peptide reversing acute and chronic morphine tolerance. AB - It has been reported that intracerebroventricular (i.c.v.) injection of nociception/orphanin FQ (OFQ) can antagonize morphine analgesia, whereas i.c.v. OFQ antibody can reverse morphine tolerance. Nocistatin (NST) is a recently characterized neuropeptide possessing an antagonizing effect on OFQ. Here we examine whether i.c.v. NST would result in a reversal of morphine tolerance. The results showed that: (1) i.c.v. NST at doses of 0.005, 0.05, 0.5, 5 or 50 ng per rat produced a bell-shaped dose-dependent reversal of chronic morphine tolerance, with maximum response at 0.5 ng. (2) Acute morphine tolerance could also be reversed, albeit partially, by i.c.v. NST at 0.5 ng. (3) The reversal of acute and chronic morphine tolerance by NST was completely abolished when NST (0.5 ng) was co-injected with (8 microg) OFQ. Since OFQ and NST are derived from the same preprohormone, the profile of its splicing in the CNS may play an important role in determining the effectiveness of morphine analgesia. PMID- 11409761 TI - High frequency evoked response to somatosensory stimulation. PMID- 11409762 TI - Hearing aid possession in the population: lessons from a small country. AB - The results of several studies on hearing aid use in the Welsh population were considered to investigate some general principles on determinants of such use within the general populations of developed countries. Overall hearing aid possession and use were not found to have changed significantly over the past 18 years, remaining at 4 per cent having obtained hearing aids and 3 per cent using them. The possession figures were consistent across all methodologies used. Higher hearing aid use in the post-industrial valleys (in which the traditional industries of coal mining and steel production had disappeared) was explained entirely by a higher level of reported hearing difficulties there. In all populations, less than 20 per cent of those reporting difficulties possessed hearing aids. Whether a hearing aid had been obtained free of charge from the National Health Service or purchased privately did not influence whether it was still used. It was concluded that an effort to improve the acceptability of hearing aids and reduce their stigma is required. PMID- 11409763 TI - Digital subtraction method for transient evoked otoacoustic emission recording with ipsilateral noise suppression: an application to stimulus artifact reduction. AB - The suppression of transient evoked otoacoustic emissions (TEOAEs) by continuous ipsilateral noise masking was investigated to explore the feasibility of its use in the elimination of the acoustical stimulus artifact. A reference noise template was obtained by stimulating the ear with identically reproducible digitally synthesized broadband noise. The same noise was used to suppress a TEOAE generated by an ipsilaterally presented click under the same conditions. Digital subtraction of the reference noise from the noise-suppressed TEOAE resulted in obtaining a template of a click artifact for that ear. The results have shown that the digital subtraction method cancels the suppressor noise, enabling the observation of the suppressed TEOAE. The subtraction of the stimulus artifact template from the original TEOAE allowed the recovery of the emission waveform with substantially reduced stimulus artifact, thus enabling the recovery of high-frequency otoacoustic emission components. PMID- 11409764 TI - Efficient stimuli for recording of the amplitude modulation following response. AB - Amplitude modulation following response (AMFR) is a promising tool for objective frequency-specific assessment of hearing thresholds in children. The stimulus generally used for AMFR recording (one amplitude-modulated carrier) activates only a small part of the basilar membrane. Therefore, the response amplitude is small. Combined frequency and amplitude modulation is not significantly more effective. A new stimulus is proposed that is composed of several carriers. All carriers are modulated with the same modulation frequency. The signal to noise ratio of the response to the multiple-carrier stimulus is significantly increased compared with the usual one-carrier stimulus. Therefore, response detection near threshold is expected to be improved. AMFR also seems to be well suited to newborn hearing screening. Simultaneous recording of six responses (three AMFRs from each ear) to 1-, 2-, and 4-kHz multiple-carrier stimuli of a 40 dB HL stimulus level is demonstrated in normally-hearing adults. Verification in babies will be necessary. PMID- 11409765 TI - Development of a central auditory test battery for adults. AB - There is little standardized test material in Dutch to document central auditory processing disorders (CAPDs). Therefore, a new central auditory test battery was composed and standardized for use with adult populations and older children. The test battery comprised seven tests (words in noise, filtered speech, binaural fusion, sentences in noise, dichotic digits, frequency and duration patterns, backward masking) and a questionnaire. Descriptive statistics obtained from 28 Dutch adults with normal hearing are presented. Eleven of the subjects were retested. The test-retest results showed that the variability between subjects was not due to systematic effects. Except for the backward masking test, the results were comparable with data from other studies. As most test scores did not follow a normal distribution, cut-off scores are recommended to evaluate individual subjects. Experiments on subjects with suspected CAPDs are in progress to validate the test battery and to see whether the number of tests can be reduced. PMID- 11409766 TI - Toward a representative set of "real-life" noises. AB - The focus of this study was to create a database containing a large number of background noises. These noises were analysed on the basis of their spectral and temporal behaviour. The most important dimensions have been determined by using a factor analysis, which reduced the total number of dimensions from 14 (7 octave bands in the spectral domain and 7 in the temporal domain) to 4. One dimension provides information about the amount of amplitude modulation present in the signal. The other dimensions provide spectral information. Based on the output of the factor analysis, a cluster analysis was carried out to reduce the total number of signals (144) to 15 anchor points (i.e., the noise forming the median of the cluster). A selection of these anchor points can be used for fitting of so called comfort programs in multi-program hearing aids and for the evaluation of signal-processing schemes in specific background noise conditions. PMID- 11409767 TI - Effect of hearing loss, centre frequency, and bandwidth on the shape of loudness functions in categorical loudness scaling. AB - Loudness functions of narrow-band and broad-band stimuli were measured with eight normally-hearing and eight hearing-impaired listeners using a categorical loudness scaling method. In the normally-hearing listeners, narrow-band stimuli, generally generated loudness functions whose slope increased with increasing level, whereas broad-band stimuli generated more linear loudness functions. These differences can be explained by the level dependence of spectral loudness summation, which is known to be most prominent at moderate levels. In the hearing impaired listeners, the narrow-band loudness functions generally showed a more linear shape than in the normally-hearing listeners. A consequence of these findings might be that the optimal shape of the input/output curve of a hearing aid is affected not only by the signal power in the respective frequency channels but also by the bandwidth of the input signal. However, there were considerable differences between listeners in both groups regarding the individual shape and absolute position of the loudness functions. Therefore, no normative reference could be extracted that would allow for a quantification of the bandwidth effect on an individual basis. PMID- 11409768 TI - Review of papers presented at the Round Table session on social and emotional development in deaf children: services in four countries. AB - In the Round Table discussion after the keynote lecture given by anthropologist Laura Lakshmi Fjord, presentations were given by Anna-Lena Tvingstedt, psychologist, PhD and research fellow, Malmo University College, Sweden; by Brita Edlund, Sign Language teacher and chair of the Finnish/Swedish association for parents of deaf children, Finland; by Dusan Kuhar, psychologist and family therapist at the Education center for deaf and hard-of-hearing children in Ljubljana, Slovenia and by Felicity Gifford, teacher of the deaf, State Coordinator of Services to Deaf and Hearing Impaired students, Department of Education, Tasmania, Australia. The focus of the session was set on needs and processes in a family when the diagnosis of a deaf child has just been given. This paper is a summary of the four presentations. PMID- 11409769 TI - Ethos and embodiment: the social and emotional development of deaf children. AB - There is so much out there about the grief, the denial, and the anger, like in the death of a child. We definitely experienced the 'death' of our child--that was not the child we gave birth to. And, we had this other baby we were going to have to figure out what to do with (mother, US, 1995). PMID- 11409770 TI - Support services in Denmark for parents of children who are deaf or hard of hearing--a national survey. AB - To collect parents' evaluation of support services in Denmark, questionnaires were distributed to parents of children who are deaf or hard of hearing. Some few results and conclusions are picked out in the article below. Parents' comments show some recurrent features: counselling must be offered and should be offered immediately after confirmed diagnosis, there is a need for psychological support at an early time, for qualified advisers, for written information to be distributed, and for co-operation between advisers of the family. PMID- 11409771 TI - Social inclusion and career development--transition from upper secondary school to work or post-secondary education among hard of hearing students. AB - The aim of this study was to investigate the decision process and to analyse the mechanisms involved in the transition from upper secondary education to post secondary education or the labour market. Sixteen students with sensorioneural hearing loss were selected. Among these eight of the students continued to university and eight did not. Twenty-five per cent of the students were women and the average age was 28 years. The investigation was conducted about 5 years after graduation from the upper secondary school. Both quantitative and qualitative methods were used. The results showed that none of the students came from a family where any or both of the parents had a university or comparable education. The differences in choice between the two groups cannot be explained in terms of social inheritance. Our study indicates that given normal intellectual capacity the level of the hearing loss seems to have no predictive value regarding future educational performance and academic career. The conclusion is that it is of great importance that a hearing impaired pupil with normal intellectual capacity is encouraged and guided to choose an upper secondary educational programme which is orientated towards post-secondary education (instead of a narrow vocational programme). Additional to their hearing impairment and related educational problems, hard of hearing students have much more difficulty than normal hearing peers in coping with changes in intentions and goals regarding their educational career during their upper secondary education. PMID- 11409772 TI - Universal newborn screening: a dream realized or a nightmare in the making? AB - There is a very strong movement to develop universal newborn hearing screening. This effort is the end product of a long international research effort to determine the most effective means to screen newborns. Now that OAE and ABR together offer a superior mechanism to achieve universal screening, problems related to middle ear effusion, non-high-risk children and adequate resources for all aspects of identification, diagnosis and treatment have come to the fore. Further, what to do in the developing world is also a major problem as audiology embarks on this exciting new frontier. This paper discusses some of the issues, raises some concerns and offers a few small solutions. PMID- 11409773 TI - The feasibility of hospital-based universal newborn hearing screening in the United Kingdom. AB - Current hearing screening programmes in the United Kingdom are performing unacceptably poorly. Davies et al. (1997) suggested that universal newborn hearing screening (UNHS) would be more effective and cheaper to run. However, there is concern that hospital-based UNHS would not be feasible because of early postnatal discharge, and thus babies not staying in hospital long enough to be screened. Two studies were designed to determine the viability of hospital-based UNHS in a district general hospital in the United Kingdom. Study 1 retrospectively determined the discharge age and time of discharge of all 3021 well babies born at St Helier hospital, Carshalton, and the number of babies born at home in the area, from 19 October 1997-18 October 1998. Most well babies were found to pass through hospital at a convenient time for predischarge hearing screening, and the optimal protocol was screening from 9 am-2 pm, 7 days a week. The predicted maximal screening coverage was 92.68%. Study 2 tested the calculated optimal protocol over 1 week. It was found that UNHS with otoacoustic emissions on the maternity ward from 9 am-2 pm, 7 days a week, achieved a coverage of 89.06%, with an acceptable false positive rate of 6.2%. It is likely that a similar protocol with slight modifications could be implemented successfully in other hospitals in the United Kingdom. PMID- 11409774 TI - Newborn hearing screening: selected experience in the United States. AB - Universal newborn hearing screening (UNHS) is rapidly becoming 'standard of care' in the United States. More than two dozen states now require, through legislative mandate, that the state establishes a system for early hearing detection and intervention (EDHI), beginning with mandated UNHS. In states with long-standing EDHI programs, the average age of identification and intervention has decreased significantly. In those states, infants are identified and intervention initiated typically before age 6 months, meeting the Joint Committee on Infant Hearing's (JCIH) recommendation for newborn hearing screening, diagnosis and intervention. Language outcome data suggest that earlier intervention results in better language outcomes for deaf and hard-of-hearing children. This article reviews the current status of UNHS in the United States, summarizes the Colorado statewide program and describes the JCIH Year 2000 position statement for developing comprehensive EDHI programs. PMID- 11409775 TI - Universal newborn hearing screening and transient evoked otoacoustic emission: new concepts in Brazil. AB - The goal of this paper is to present a Brazilian universal newborn hearing screening (UNHS) program based on transient evoked otoacoustic emission (TEOAE) and study the hearing impairment prevalence in this population. From September 1996 to August 1999, 4631 babies were born at the maternity ward of Hospital Israelita Albert Einstein in Sao Paulo, Brazil and 4196 (90.6%) had a hearing screening test performed before discharge. The equipment ILO88, 'quickscreen' mode and the 2 stages protocol were used. From the 4196 babies tested, 4123 (98.2%) had a normal test and 73 (1.8%) failed at the first stage screening. The follow-up was performed in 60 (82%) of those 73 babies and 10 (2.3-1000 live births) had a confirmed hearing loss, three without hearing risk factors. The TEOAE UNHS program was a feasible and accurate method to detect hearing disorders at an early stage. PMID- 11409776 TI - Normal values for distortion product otoacoustic emissions in children: a study using primary levels previously demonstrated to be optimum for identification of hearing loss. AB - No published data exist for normal values of distortion product otoacoustic emissions (DPOAE) in children at primary levels f1 = 65 dB and f2 = 55 dB SPL. These primary levels have been previously demonstrated to be optimal for identification of hearing impaired ears in adults. A total of 102 normal children underwent audiological assessment, including exclusion of middle ear disease, pure tone audiometry and DPOAE DP-grams (primaries L1/L2 = 65/55 dB SPL, f1:f2 = 1.22). There was a statistically significant decrease in DPOAE amplitude with increasing age. DPOAE amplitude was also dependent on the frequency of f2. However, there was wide inter- and intra-individual variation in DPOAE amplitude at different frequencies of f2. There was also a large overlap between the range of values of DPOAE amplitude between the adjacent age groups. Detailed assessment of DPOAE in children is feasible in the clinical setting. These normal values should prove invaluable in future studies; however, the large range of normal values means that cross-sectional studies may not be able to detect small variations in cochlear function. PMID- 11409777 TI - Genes in the ear: what have we learned over the last years? AB - In developed countries 50% of childhood hearing impairment is attributable to genetic causes. In a limited number of cases, the hearing impairment is part of a syndrome, and several genes for syndromic deafness have been identified over the last 10 years. In the majority of cases, the hearing impairment occurs without additional clinical abnormalities (non-syndromic). Progressive hearing loss is very frequent in adults. By the age of 80 approximately 50% of the population is affected by age-related hearing loss, which is due partly to genetic factors. Before 1994, little was known about the genes responsible for non-syndromic hearing impairment, although epidemiological studies have suggested that more than 100 genes might be involved. Over the last 6 years, extremely rapid progress was realized in the field of the molecular genetics of hearing and deafness. More than 70 genes for non-syndromic hearing impairment have been localized to the human genome, and 18 of these have been identified. PMID- 11409778 TI - Children's amplification needs--same or different from adults? AB - Hearing aid fitting strategies for children have largely developed separately from those for adults. There has, however, been little consideration of whether children's amplification requirements are different, except for the physical effects of differences in ear canal size. This paper summarizes the fitting strategies for children, and examines whether children require more (or less) gain for high-, medium- or low-level sounds, or a different frequency response, compared with adults with similar hearing loss. Research relating to the above questions is reviewed. The gain that children require for high-level sounds is not different than for adults because loudness discomfort levels are similar. The gain children prefer for medium-level sounds is the same, or only slightly more, than is preferred by adults. Children may require greater gain for low-level sounds because they do require higher signal levels to achieve the same level of speech understanding as adults do. There are, however, arguments why more gain for low-level sounds may not be desirable. The frequency response children prefer is the same as that preferred by adults. New methods for evaluating individual fittings are briefly discussed. PMID- 11409779 TI - Auditory performance of young children with hearing aids: the Nijmegen experience. AB - During recent years, significant progress has been made in the development of methods for screening hearing sensitivity in toddlers. Nowadays, in several countries, nationwide screening programmes are carried out. Much less attention has been paid to the next step, namely treatment of very young hearing-impaired children with hearing devices. A review of published data indicate that the DSL (Desired Sensation Level) hearing aid prescription method, developed especially for young children, is at least a good point to begin hearing aid fitting. DSL target values for gain and output seems to be adequate for young children. Documenting the benefit for hearing aid use in young children is still cumbersome. One option is to measure the development of basal auditory functions. A review of available data showed that for this purpose, speech material can only be used in children of 3 years and older. For children of 1 year and older, tests with environmental sounds can be applied. It is concluded that, although growing, there is a lack of tests and limited knowledge on how to optimize hearing aid fittings and how to document hearing aid benefit in toddlers. PMID- 11409780 TI - How to assess outcome of hearing aid fitting in children. AB - Assessment of outcome of hearing aid fitting in children should contain several dimensions: audibility, speech recognition, subjective benefit and speech production. Audibility may be determined by means of aided hearing thresholds or real-ear measurements. For determining speech recognition, methods different from those used for adult patients must be used, especially for children with congenital hearing loss. In these children the development of the spoken language and vocabulary has to be considered, especially when testing speech recognition but also with regard to speech production. Subjective assessment of benefit to a large extent has to rely on the assessment by parents and teachers for children younger than school age. However, several studies have shown that children from the age of around 7 years can usually produce reliable responses in this respect. Speech production has to be assessed in terms of intelligibility by others, who may or may not be used to the individual child's speech. Ideally, the outcome should be assessed repeatedly at suitable intervals in order to be able to follow the child's communicative development. PMID- 11409781 TI - The effects of age of cochlear implantation on speech perception outcomes in prelingually deaf children. AB - We have made a retrospective analysis on 70 prelingually deaf children (78% congenital; age range 2-15 years) followed for up to 5 years post-implant during which both closed set speech perception tests (TAC, WIPI) and open set tasks (PBK, GASP) were administered. We used a binary partitioning algorithm to optimally divide our dataset on the basis of age at implantation This technique achieves an optimal split when the heterogeneity of the data is most reduced (maximal drop in deviance). For the closed set speech perception tests (TAC and WIPI) partitioning best divided-out data at age 4.4 years. For the open set tests optimal division was at a higher age of implantation (GASP word, 5.6 years; PBK word, 8.4 years). Using these partitioning values, we have found statistically significant differences between rate of improvement of scores in the younger implanted children compared with those implanted later. PMID- 11409782 TI - Phonological development in 4-6-year-old moderately hearing impaired children. AB - The acquisition of clear phonological rules is of utmost importance for a child when he/she is learning to decode and convey meanings using spoken language. Fifteen Finnish-speaking children, five normally hearing (NH) 3-year-olds and 10 moderately hearing impaired (HId) 4-6-year-olds were studied here. Speech samples consisting of 62 words were collected from each child during a picture naming task. Frequent phonetic (articulation) errors together with normal (but delayed) and deviant phonological processes of HId children were found in this study. The frequency and quality of the processes seemed to be related more to the grade of HI than, for example, the age of the child. Implications for speech therapy are briefly discussed. PMID- 11409783 TI - Age-related tonotopic map plasticity in the central auditory pathways. AB - Inner hair cell lesions to the basal turn of the cochlea effectively result in a partial deafferentation of the auditory system. At the level of the midbrain (central nucleus of inferior colliculus) cochleotopic maps, based on single unit response characteristic frequency, are changed after such deafferentation. When a cochlear lesion is induced in a neonatal animal (chinchilla), the reorganization of the frequency map is more extensive than that resulting from similar deafferentation in the adult subject. Neonatal cochlear lesions result in an over representation of sound frequencies corresponding to the border of the cochlear lesion, while similar lesions in the adult do not. The results suggest that significant plasticity exists in the auditory midbrain during early post natal development (even in a precocious species, such as chinchilla); however, this plasticity is largely lost in the mature animal. A conceptual model for the frequency map re-wiring is presented. PMID- 11409784 TI - Hearing impaired children in developing countries. AB - The majority of hearing impaired children live in the developing countries. Limited resources are available for screening, prevention, diagnosis and intervention. These problems were the topic for discussion in a special session in the Congress. PMID- 11409785 TI - Audiology in Latin America: hearing impairment, resources and services. AB - Evidence is presented about the limitation of information available on prevalence/incidence of deafness and hearing impairment in the developing world; particularly in Latin America. Two questionnaires on audiological resources and services were mailed to Latin American and Caribbean countries in general and to Central American nations in particular. The information returned by Argentina, Chile, Colombia, Costa Rica, Cuba, Belize, El Salvador, Grenada, Guatemala, Honduras, Mexico, Nicaragua, Panama, Puerto Rico and Uruguay was analysed. Data was gathered about epidemiological studies on hearing impairment, about specific programmes, such as early identification of hearing impairment, national registers on deafness and programmes on hearing screening. Training programmes and availability of professionals in the field of hearing and deafness is also shown. Hearing services, hearing aids provision, hearing testing equipment, professional organization and legislation in audiology is also documented. It is our conclusion that hearing impairment is a low priority for health systems in the developing world, technology continues to be excessively costly and material and human resources are limited, and services are poor and restricted. PMID- 11409786 TI - WHO activities for prevention of deafness and hearing impairment in children. AB - This paper gives an overview of WHO activities in the global campaign for the prevention of deafness and hearing impairment, focusing particularly on children. It discusses the size of the problem and the causes and consequences of deafness and hearing impairment. It emphasizes the inadequate state of our knowledge of this subject in developing countries and the importance of collecting valid data. It describes the public health route to prevention of deafness and hearing impairment, especially through primary ear and hearing care, and outlines the World Health Organization's aims and activities for prevention of deafness and hearing impairment. PMID- 11409787 TI - Characteristics of the audiometric notch as a clinical sign of noise exposure. AB - This study investigated variability in audiometric notch recognition by asking three clinicians to identify notches in the audiograms of 634 noise-exposed employees. Agreement between the raters was assessed, and three notch characteristics, frequency, depth and shape (a wide or narrow notch), were investigated as recognition factors. The proportions of positive tests reported by each rater were 0.26, 0.49 and 0.68, respectively. The 'all rater' index of agreement was 0.45, and the range for pairwise comparisons was 0.14 to 0.52. With recognition factors, rater 1 was most strongly influenced by depth, rater 2 by shape and rater 3 by a possible combination of criteria. To reduce variability we suggest that narrow notches should be at least 15 dB in depth, and that broad notches should have a depth of 20 dB, with a recovery of at least 10 dB at the high end. Care should also be taken to elicit a sufficient history of noise exposure, and ensure that there are no other explanations for the notch. PMID- 11409788 TI - Speech perception and functional benefit after cochlear implantation: a multicentre survey in Finland. AB - This study was done to survey the effect of cochlear implantation on hearing level, speech perception and listening performance in Finnish-speaking adults. The subjects of the study comprise 67 adults. Pure-tone thresholds (0.125-8 kHz), word recognition and listening performance were studied before and after implantation. After switch-on of the implant, the median values of PTA(0.5-4 kHz) in the sound field were fairly stable across the evaluation period. Three months after switch-on of the implant, the mean word recognition score was 54%. There was clear improvement in the mean word recognition scores over a longer period of time, the mean score being 71% 24 months after switch-on. Six months after switch on, the majority of subjects (40/48) were able to recognize some speech without speechreading, and 26 of these 48 subjects were able to use the telephone with a known speaker, gaining good functional benefit from the implantation. PMID- 11409789 TI - Loudness perception and late auditory evoked potentials in adult cochlear implant users. AB - In clinical routine the adjustment of speech processors in cochlear implant users is based on the patients' subjective statements about the loudness of specific electrical stimuli. From hearing patients it is known that the latencies and amplitudes of late auditory evoked potentials (LAEP) which are generated within the auditory cortex correlate with the loudness perception of acoustical stimuli. The aim of this study was to investigate the correlation between LAEP and loudness perception in adult cochlear implant users. We investigated 8 adult subjects who had been provided with a 22 electrode Cochlear Implant (nucleus CI24M) at least 6 months prior to the investigation. All subjects showed open speech understanding. Electrical pulse trains of 300 ms duration presented at a single electrode served as stimuli. Electrically LAEP morphologies were similar to normal hearing subjects. In all subjects and each intracochlear electrode position LAEP were well identifiable down to low loudness sensations. Both amplitudes and latencies depended on the loudness perception. The best correlation was observed for the N1 deflection. The results show that LAEP can be used for estimation of both hearing thresholds and most comfortable levels. PMID- 11409790 TI - A review and evaluation of research on the deaf-blind from perceptual, communicative, social and rehabilitative perspectives. AB - This paper reviews research on deaf-blind individuals, primarily from behavioral and communicative points of view. Inclusion in the population of deaf-blind is qualified by describing a variety of subgroups and genetically based syndromes associated with deaf-blindness. Sensory assessment procedures--based primarily on residual capacities--are appraised. Consequences for everyday life are described briefly. Non-sensory, alternative classificatory schemes and procedures are presented and the results from behavior modification procedures used for correcting maladaptive behaviors are summarized. Methods for communicating tactilely are described and evaluated. Attention is also drawn to some suggestions regarding learning of alphabetic codes and sign acquisition. Finally, suggestions for future research are proposed. PMID- 11409791 TI - Objective detection of transiently evoked otoacoustic emissions. AB - Newborn hearing screening with transiently evoked otoacoustic emissions (TEOAE) is a well-established method. A screening device must be equipped with a test procedure for objective TEOAE detection. The statistical tests implemented in the commercially available screening devices are the correlation, an estimation of the signal-to-noise ratio and a binominal test. The aim of the present study is to compare the TEOAE detection performance of these tests with that of several other tests in the time and frequency domains (variance ratio F(SP) and its modification F(SP)*, Friedman test, modified q-sample uniform scores test). The comparison was based on a data sample of 420 TEOAE. The frequency range examined was 1.5-4.0 kHz. As a new feature, two frequency sub-ranges (1.5-2.5 kHz, 2.5-4.0 kHz) were tested separately. The modified variance ratio F(SP)* was the most powerful test, whereas the tests implemented in the known screening devices showed the lowest detection performance. PMID- 11409792 TI - Speech-reading of synthetic and natural faces: effects of contextual cueing and mode of presentation. AB - A natural and a synthetic face were compared with regard to speech-reading performance, with a visual and an audio-visual condition, and with three levels of contextual cueing in an experiment with 90 normal-hearing subjects. Auditory presentation (speech in noise) served as a control condition. The results showed main effects for type of face, presentation mode and contextual cueing, and also an interaction between presentation mode and type of face, such that performance was superior for the natural compared with the synthetic face, and especially in the audio-visual mode. Audio-visual speech-reading was more accurate than visual and auditory presentation and contextual cueing improved performance overall. It is suggested that a synthetic face can be a valuable and useful support if proper cueing and an auditory signal are provided, but it is outperformed by a natural face with regard to complementarity to an underspecified auditory speech signal. PMID- 11409793 TI - Can maximum comfortable loudness levels in hearing impaired listeners be predicted from ipsilateral acoustic reflex thresholds recorded with high frequency probes? AB - This study was designed to test the effectiveness of the ipsilateral acoustic reflex thresholds measured with low and high frequency probes in predicting the maximum comfortable loudness levels (MaxCLs) in hearing-impaired subjects using recently available sophisticated procedures for acoustic reflex and loudness measures. Loudness growth functions were measured in hearing-impaired subjects at .5, 1 and 2 kHz using a computerized procedure. The maximum level among those sound pressure levels judged as 'comfortable' was designated as the MaxCL. Acoustic reflex thresholds were measured for .5, 1 and 2 kHz activator signals using 226, 678 and 1000 Hz probes. Regression analyses suggested that acoustic reflex thresholds recorded with the 678 and 1000 Hz probes could provide an objective estimate of the MaxCLs for 1 and 2 kHz. Thus, acoustic reflex thresholds may be useful in fitting hearing aids for those subjects in whom reliable measures of MaxCLs cannot be obtained. PMID- 11409794 TI - The future of surgery and surgical organizations. PMID- 11409795 TI - The history of the Gold Medal Papers of the Southeastern Surgical Congress: 1960 2000. AB - Since 1960 at its annual meeting the Southeastern Surgical Congress has selected a varying number of "Gold Medal Papers" to be presented. Customarily a prize is awarded for the first place among the Gold Medal Papers and in some instances a second and third place. A brief history of the origin of these Gold Medal Papers is presented. With the assistance of the Executive Secretary of the Southeastern Surgical Congress a list of the Gold Medal presenters insofar as can be ascertained from the Council minutes is presented. It is unfortunate that all of the names of the Gold Medal Paper presenters have not been preserved. PMID- 11409796 TI - Polymerase chain reaction for the detection of bacteremia. AB - Analysis of blood by polymerase chain reaction (PCR) is a more rapid and sensitive method to detect bacteremia than blood culture. The PCR was performed on blood obtained from patients during blood culture draws and on blood from normal volunteers. Eighty-seven patients provided 125 blood samples for blood culture comparison with PCR. Specific PCR primers for Staphylococcus aureus and Escherichia coli that targeted conserved regions common to gram-positive and gram negative bacteria were used. Selective stringency conditions identified other gram-positive and gram-negative bacteria. The blood culture agreed with the PCR in 93 of the 125 patient specimens (74%). In 29 of these specimens the PCR was positive yet the blood culture was negative. When clinical information was included with positive blood culture to define sepsis in these patients and their specimens were added to the positive blood cultures the statistical accuracy of PCR was 93 per cent. Only three of the 78 specimens with negative PCR had positive blood cultures. The PCR was negative in all but one of the 50 volunteers. PCR is more sensitive than blood culture, and it can quickly rule out bacteremia. PMID- 11409797 TI - Importance of lymphatic mapping in ductal carcinoma in situ (DCIS): why map DCIS? AB - The appropriateness of sentinel lymph node biopsy in the management of patients with biopsy diagnoses of ductal carcinoma in situ (DCIS) or DCIS with microinvasion (DCISM) has not been established. Three hundred forty-one patients presented with a biopsy diagnosis of DCIS or DCISM. Two hundred forty (70%) underwent sentinel node biopsy at their definitive procedure. All clinical and pathologic data were collected prospectively. Of 224 patients with a biopsy diagnosis of DCIS 23 (10%) were upstaged to infiltrating ductal carcinoma (IDC) at their definitive therapy and of 16 patients with a biopsy diagnosis of DCISM seven (44%) were upstaged to IDC. Excisional biopsies were no more sensitive for detecting IDC than was core biopsy. Lymph node metastases were detected in 26 of 195 (13%) patients with a definitive diagnosis of DCIS, in three of 15 (20%) with a definitive diagnosis of DCISM, and in eight of 30 (27%) with a definitive diagnosis of IDC. Sentinel lymph node biopsy is a valuable tool in the treatment of patients with DCIS and DCISM and is particularly needed in those undergoing mastectomy. No "high-risk" group of patients can be identified for selective sentinel lymph node biopsy. PMID- 11409798 TI - Accuracy of sentinel lymph node biopsy for patients with T2 and T3 breast cancers. AB - Although numerous studies have demonstrated that sentinel lymph node (SLN) biopsy can accurately determine the axillary nodal status for early breast cancer some studies have suggested that SLN biopsy may be less reliable for tumors >2 cm in size. This analysis was performed to determine whether tumor size affects the accuracy of SLN biopsy. The University of Louisville Breast Cancer Sentinel Lymph Node Study is a prospective multi-institutional study involving 226 surgeons. The study was approved by the Institutional Review Board of each institution, and informed consent was obtained from all patients. Patients with clinical stage T1 2 N0 breast cancer were eligible for the study. Some patients with T3 tumors were included because they were clinically staged as T2 but on final pathology were found to have tumors >5 cm. This analysis includes 2148 patients who were enrolled from August 1997 through October 2000. All patients underwent SLN biopsy using a combination of radioactive colloid and blue dye injection followed by completion Level I/II axillary dissection. Statistical comparison was performed by chi-square analysis. The SLN identification rate, false negative rate, and overall accuracy of SLN biopsy were not significantly different among tumor stages T1, T2, and T3. We conclude that SLN biopsy is no less accurate for T2-3 breast cancers compared with T1 tumors. PMID- 11409799 TI - Clinical utility of frozen section in sentinel node biopsy in breast cancer. AB - One hundred sixty-five breast cancer patients underwent a sentinel lymph node biopsy procedure over a period of 2 years. Sentinel node (SN) could be successfully localized in 157 (95%) of the patients. Complete axillary lymph node dissection was performed only if the frozen section (FS) revealed a positive SN. All SN specimens were further evaluated by hematoxylin and eosin on multiple sections and cytokeratin immunohistochemisty. The patients whose SNs were negative by FS but positive by permanent histopathologic evaluation underwent a delayed axillary lymph node dissection. SN was positive in 41 of 157 (26%) patients. Eighteen (44%) of the 41 patients with SN metastases were diagnosed intraoperatively by FS and underwent a one-stage definitive surgical treatment. The benefit of FS was most notable in patients with T1c and larger lesions. PMID- 11409800 TI - Does intraoperative electrohydraulic lithotripsy improve outcome in the surgical management of chronic pancreatitis? AB - Lateral pancreaticojejunostomy (LPJ) is the cornerstone of surgical management of pain associated with chronic pancreatitis (CP) and ductal dilation. The pathologic key to failure of LPJ is disease confined to the head of the pancreas. Intraoperative pancreatoscopy with electrohydraulic lithotripsy (EHL) is a novel technique that avoids resection and eradicates intraductal lithiasis in the head of the gland. This study was undertaken to compare outcome of LPJ alone and LPJ with intraoperative EHL in the surgical management of CP. The records of patients undergoing LPJ with intraoperative EHL between 1996 and 1998 (Group A) were reviewed and compared with our historical data of patients who underwent LPJ alone from 1977 through 1991 (Group B). Quality-of-life questionnaires were administered in person or by telephone. Fisher's exact and Mann-Whitney statistical tests were used where appropriate. Twenty patients (12 men, 8 women; mean age 51 years, range 29-68) in Group A underwent LPH with EHL versus 85 patients in Group B (65 men, 20 women; mean age 43.6 years, range 24-73) who had LPJ only. The etiology of CP was attributed to alcohol abuse in 85 per cent of patients in Group A and 96 per cent in Group B. Mean follow-up for Group A was 2.7 years (range 1-4 years) and 6.3 years (range 1-15 years) for Group B. Complications occurred in four patients (Group A) and five patients (Group B) perioperatively. There were no deaths in either group in the early postoperative period. Subsequent operations for complications of CP were significantly fewer in Group A than in Group B (P < 0.05). Rehospitilizations were required in 35 and 60 per cent of patients in Group A and B respectively (P < 0.05). Postoperative insulin and enzyme supplementation requirements were unchanged in Group A and continued or worsened in Group B. Ninety per cent of patients in Group A viewed their health status as good or fair compared with 55 per cent in Group B (P < 0.05). Postoperative narcotic use was present in both groups, although the number of pain pills used decreased considerably from 25 per week to fewer than five in Group A. Intraoperative EHL may represent an alternative to resection of the head of the pancreas or may be used as an adjunct to LPJ in the surgical management of chronic fibrocalcific pancreatitis. PMID- 11409801 TI - Delay in treatment of biliary disease during pregnancy increases morbidity and can be avoided with safe laparoscopic cholecystectomy. AB - Recent reports indicate that laparoscopic cholecystectomy in pregnancy is safe. The aim of this study was to evaluate whether delays in definitive treatment of symptomatic cholelithiasis increase morbidity. We reviewed the records of 16 women who underwent laparoscopic cholecystectomy during pregnancy between 1992 and 1999. Mean age was 24 +/- 5 years (mean +/- standard error). Symptom onset was during the first trimester in nine patients, second trimester in six patients, and third trimester in one patient. Patients had abdominal pain (93%), nausea (93%), emesis (80%), and fever (66%) for a median of 45 days (range 1-195 days) before cholecystectomy. Nine of 11 women who underwent cholecystectomy more than 5 weeks after onset of symptoms experienced recurrent attacks necessitating 15 hospital admissions and four emergency room visits. Moreover four women who developed symptoms in the first and second trimesters but whose operations were delayed to the third trimester had 11 hospital admissions and four emergency room visits; three of those four (75%) women developed premature contractions necessitating tocolytics. Cholecystectomy was completed laparoscopically in 14 women. There was no hospital infant or maternal mortality or morbidity. We recommend prompt laparoscopic cholecystectomy in pregnant women with symptomatic biliary disease because it is safe and it reduces hospital admissions and frequency of premature labor. PMID- 11409802 TI - Magnetically guided nasoenteral feeding tubes: a new technique. AB - Our objective was to evaluate a new technique for the bedside placement of nasoenteral feeding tubes into the duodenum using an external hand-held magnet to maneuver the tube from the stomach to the distal duodenum. We conducted a prospective case series of 20 consecutive patients requiring nasoenteral tube feeding in the intensive care units of a university-affiliated hospital. Twenty patients were entered into the study after the attending physician requested assistance in tube placement. A flexible nasoenteral feeding tube (12 F), modified to include a magnet and a magnetic field sensor in the distal tip connected by a thin insulated wire to a small light at the proximal end, was passed per nares into the stomach. A larger hand-held magnet held over the epigastrium was used to magnetically "capture" the tube tip, indicated by the illumination of the proximal light. The tube tip was then maneuvered by the hand held magnet along the lesser curvature of the stomach, through the pylorus, and into the duodenum. Procedure time and anatomic location of the tube tip as determined by an abdominal radiograph was recorded. The 12 men and eight women had a mean age of 60 years (range 30-84). The procedure time averaged 9.6 minutes (range 1-30). In 19 of the 20 patients (95%) radiographs revealed successful placement of the tip of the feeding tube into the duodenum. There were no complications related to the procedure. Using a novel magnetically guided nasoenteral feeding tube transpyloric tube placement was achieved in 95 per cent of cases with an average procedure time of 9.6 minutes. This new and inexpensive bedside technique will allow prompt and safe initiation of enteral nutrition. PMID- 11409803 TI - Blunt renal artery injury: incidence, diagnosis, and management. AB - Renal artery injury is a rare complication of blunt abdominal trauma. Increasing use of CT scans to evaluate blunt abdominal trauma identifies more blunt renal artery injuries (BRAIs) that may have otherwise been missed. We identified patients with BRAI to examine the incidence and to evaluate the current diagnosis and management strategies. Patients admitted from 1986 to 2000 at a regional Level I trauma center sustaining BRAI were evaluated. Patients undergoing revascularization or nonoperative management were followed for renovascular hypertension. Twenty-eight patients with BRAI were identified out of 36,938 blunt trauma admissions between 1986 and 2000 (incidence 0.08%). Most renal artery injuries were diagnosed by CT scans (93%) with seven confirmatory angiograms. Nine patients had nephrectomy (one bilateral), and three patients with unilateral injuries were revascularized. Sixteen were managed nonoperatively including one patient who had endovascular stent placement. Three patients died from shock and sepsis. Follow-up for all patients ranged from one month to 8 years. Two patients developed hypertension: one who was revascularized (33%) and one was managed nonoperatively (6%). The frequency of diagnosis of BRAI is increasing because of the increased use of CT. Nonoperative management of unilateral injuries can be successful with a 6 per cent risk for developing renovascular hypertension. The role of endovascular stenting is promising, and further study is necessary. PMID- 11409804 TI - Bile duct injury following laparoscopic cholecystectomy: a cause for continued concern. AB - Previous reports suggest that bile duct injuries sustained during laparoscopic cholecystectomy (lap chole) are frequently severe and related to cautery and high clip ligation. We performed a review of patients who sustained bile duct injury from lap chole since 1990 and assessed time to injury recognition, time to referral, Bismuth classification, initial and subsequent repairs, rate of recurrence, and length of follow-up. Seventy-four patients [median age 44 years, 58 of 74 female (78%)] were referred with a bile duct injury after lap chole. The level of injury was evenly divided between the bile duct bifurcation and the common hepatic duct: Bismuth III, IV, and V (40 of 74, 54%) versus Bismuth I and II (34 of 74, 46%). Concomitant hepatic arterial injury was identified in nine (12%) patients. Patients referred early after bile duct injury and requiring operative intervention underwent hepaticojejunostomy at a median of 2 days after referral. After surgical reconstruction at our center there has been an overall success rate of 89 per cent with no need for reintervention. Six (10%) of these patients have required one additional balloon dilatation at a mean follow-up of >24 months. One (2%) patient underwent biliary-enteric revision in follow-up. In patients with bile duct injury, stricture repair without delay was successful in the majority of patients treated in this series. Only one of 64 patients reconstructed at our center has required reoperation; six others have required a single balloon dilatation with subsequent good or excellent results. The majority of patients treated with operative repair at an experienced center can expect good long-term results with rare need for reintervention. PMID- 11409805 TI - Results with abdominal vascular trauma in the modern era. AB - This is a report of a 10-year experience (1989-1998) with 300 consecutive patients found to have an injury to a named abdominal vessel at the time of an exploratory laparotomy for trauma. An abdominal gunshot wound was the mechanism of injury in 78 per cent of patients, and injury to more than one named abdominal vessel was present in 42 per cent. The abdominal aorta, inferior vena cava, and external iliac artery and vein were the most commonly injured vessels. When management for the five most commonly injured arteries was grouped, exsanguination before attempts at repair occurred in 11 to 15 per cent of patients and the mean survival in the remainder was 46 per cent. When management for the five most commonly injured veins was grouped, exsanguination before attempts at repair occurred in 5 per cent of patients and the mean survival in the remainder was 64 per cent. A number of administrative and medical changes in the management of patients with abdominal trauma occurred from 1992 through 1994. Despite significantly increased Injury Severity Scores for patients treated from 1993 through 1998 as compared with those treated from 1989 through 1992 survival rates for patients with injuries to the abdominal aorta and inferior vena cava were unchanged. Survival rates for injuries to the external iliac artery and vein increased significantly. The local changes in management should be considered for prospective studies in other urban trauma centers. PMID- 11409806 TI - Stereotactic breast biopsy: a study of first core samples. AB - Stereotactic core needle biopsy (SCNB) is a sensitive and specific indicator of breast pathology. Commonly the first biopsy core is taken from the center of the lesion in question. Multiple cores are then taken from points peripheral to the central core. The sensitivity and specificity of the central core to diagnose breast disease is unclear. We compared the pathology of the central core biopsy with that of the remaining cores in a prospective study to determine the sensitivity and specificity of the central core to diagnose breast disease. All patients undergoing SCNB for breast lesions in a single surgical office during a 7-month period were eligible for inclusion. One hundred thirty-three patients with first cores from 145 biopsy sites were included. The histologic diagnosis from 117 (81%) of the first cores from these 145 biopsy sites were representative of their respective samples as a whole. Seventy-seven (53%) of the first cores were in complete agreement with the final histologic diagnosis whereas 40 (28%) had minor differences with the histologic diagnosis that had little or no clinical significance. Twenty-eight (19%) central core samples did not agree with the final pathologic diagnosis. Seven of these 28 patients each had a final diagnosis of cancer missed by the central core biopsy. The first core sample had a sensitivity for cancer detection of 79 per cent and specificity 100 per cent. SCNB remains a sensitive and specific identifier of breast pathology. When mammographic evidence of calcifications was the primary indication for SCNB (n = 75) calcification was present in the central core in 51 (68%). In these 51 patients the central core biopsy was in agreement with the final histologic diagnosis in 46 (90%) specimens. Histologic review of the first core sample alone lends no increased benefits and in fact misrepresents the pathology present in a significant number of patients. When analyzed as an independent predictor of breast pathology the first core is a more sensitive indicator than subsequent individual cores, but the most accurate predictor of pathology is examination of the entire group of core samples. This study confirms the need for acquisition of multiple cores from each lesion in question. PMID- 11409807 TI - Solitary fibrous tumors arising in abdominal wall hernia sacs. AB - Solitary fibrous tumor (SFT) of the peritoneum is an unusual spindle-cell neoplasm. SFT was originally described in the pleura; however it is now diagnosed in multiple extrathoracic sites. Most believe that the tumor is of mesenchymal origin and should be classified as a variant of fibroma. SFT of the pleura and peritoneum have also been called fibrous mesothelioma, and the cell of origin is felt to be a pluripotential submesothelial mesenchymal cell. Primary tumors arising in hernia sacs are rare, and we report on two patients with hernia SFT. The first is a 67-year-old man who had a diffusely thickened distal left inguinal hernia sac. Within the sac was copious myxoid material mimicking pseudomyxoma peritonei. Herniorrhaphy and orchiectomy were performed. The second is a 44-year old woman with a midepigastric mass attached to a ventral hernia. Wide local excision was performed. Both tumors demonstrated plump spindle cells, one with myxoid background and the other with keloidal collagen. Calretinin immunostaining was positive in both tumors, whereas CD34 was negative. This suggests tumor origin from a submesothial pluripotential cell that maintains potential for mesothelial differentiation. Surgical excision is the treatment of choice with the degree of resectability being a powerful predictor of outcome. PMID- 11409808 TI - Incidental thyroid carcinoma identified by positron emission tomography scanning obtained for metastatic evaluation. AB - Positron emission tomography (PET) scanning is an emerging technology with substantial implications for patients with solid tumors. PET is useful in identifying sites of metastatic disease as well as primary malignancy and evaluating suspicious findings found via other imaging modalities. PET is becoming a standard modality for metastatic evaluation of a variety of malignancies. However, this imaging is not limited by histology and may reveal clinically occult second lesions. We present five cases of a clinically occult thyroid cancer found in patients who underwent PET scanning for metastatic evaluations. The principal diagnoses included two patients with malignant melanoma, one each with gastric, rectal, and colonic adenocarcinoma. None of the cases had findings on physical examination. Increased uptake in the region of the thyroid was evident on the PET scan, and fine-needle aspiration biopsy indicated papillary carcinoma of the thyroid in all five cases. Although evaluation of metastatic disease is the primary purpose for PET in this setting the discovery of occult thyroid malignancy is an additional benefit of such imaging. This emphasizes the importance of investigating suspicious areas found by PET and not simply assuming that these are all sites of metastatic disease. Furthermore incidental findings of uptake in the region of the thyroid make consideration of a primary carcinoma mandatory. PMID- 11409809 TI - Paraneoplastic hypercalcemia in a patient with adenosquamous cancer of the colon. AB - Hypercalcemia is a well-known manifestation of paraneoplastic syndromes associated with a variety of malignancies. However, colon cancer has only rarely been associated with hypercalcemia of malignancy. We present the case of a patient with recurrent adenosquamous carcinoma of the ascending colon found to have hypercalcemia. The patient is a 76-year-old white woman who initially presented with colon cancer in the cecum and underwent a right hemicolectomy. All lymph nodes and surgical margins were free of tumor. Pathological examination at that time revealed adenosquamous carcinoma of the colon. Eight months later she complained of dizziness, anorexia, and constipation and was found to have a calcium level of 13.6 mg/dL. CT scan revealed a mass measuring 10.5 to 12.7 cm in the right hepatic lobe, and a bone scan was normal. Her intact parathyroid hormone (PTH) level was 6 pg/mL (normal 12-72) and her PTH-related protein (PTHrP) level was 25.7 pmol/L (normal <1.3). She then underwent a hepatic resection. The serum PTH, calcium, and PTHrP levels normalized after resection. Hypercalcemia of malignancy in colon cancer is rare and has an association with adenosquamous histology. The hypercalcemia is attributed to PTHrP, and here we demonstrate this in the serum and tumor specimens. The effects of PTHrP are shown to be short-lived postoperatively. We find only 14 other cases in the literature of hypercalcemia related to a colonic neoplasm, and this is the only patient reported to be surviving. The diagnosis of a paraneoplastic syndrome mediated via PTHrP should be considered when hypercalcemia is encountered in the setting of metastatic colon carcinoma. PMID- 11409810 TI - Chlamydia pneumoniae in atherosclerotic carotid artery plaques: high prevalence among heavy smokers. AB - This study was designed to determine the prevalence of Chlamydia pneumoniae in carotid artery plaques. Although there have been numerous studies evaluating coronary plaques for this bacterium fewer studies have assessed noncoronary vasculature. In addition we wished to evaluate whether correlation exists between the presence of C. pneumoniae in carotid plaques and established risk factors for atherosclerosis. Sixty intact carotid artery plaques removed during surgery (carotid endarterectomy) were formalin-fixed and paraffin-embedded according to conventional techniques. These samples were evaluated by polymerase chain reaction analysis to detect presence of C. pneumoniae DNA. Results were tabulated and compared against established risk factors for atherosclerosis: diabetes, hypertension, hyperlipidemia, age, and smoking. Forty-two (70.0%) of the 60 plaques that were evaluated tested positive for the presence of C. pneumoniae DNA by polymerase chain reaction analysis. In the sample defined as being from heavy smokers (greater than 15-pack-year history) 33 (94.3%) of 35 plaques tested positive whereas two (5.7%) tested negative. This correlation demonstrated statistical significance (P = 1.36 x 10(-6), two-tailed Fisher exact test). Presence of C. pneumoniae in carotid plaques demonstrated no statistically significant correlation with diabetes, hypertension, or hyperlipidemia. Age as a risk factor was examined but not statistically evaluated because of the narrow range within our patient sample. Analysis of the data reveals that C. pneumoniae is present in large numbers of atheromatous plaques as is consistent with emerging data. What is interesting though is that 33 (94.3%) of the 35 smokers had plaques that tested positive for the bacterium as opposed to only nine (36.0%) of the 25 nonsmokers. Identification of specific populations exhibiting a high prevalence of C. pneumoniae may serve to focus future studies. Ongoing investigation will seek to determine whether C. pneumoniae plays an active role in the pathogenesis of atherosclerosis. PMID- 11409811 TI - Mediastinal impalement with survival: a case report. AB - Mediastinal impalement injuries are rare and often fatal. Very few instances of survival after mediastinal impalement have been reported. We present the unusual case of an 18-year-old man who was involved in a motor vehicle crash in which a wooden fencepost intruded through the windshield and impaled him through the superior mediastinum. The patient remained hemodynamically stable and had no other significant injuries except a left pneumothorax. Arteriogram revealed a bovine aortic arch with the wooden piece passing over the aortic arch between the two brachiocephalic arteries at the precise point that a normal left common carotid artery would have been located. No other injuries were seen on arteriogram, venogram, or esophagram. The foreign body was extracted via thoracotomy along with resection of the apex of the left lung and ligation of the thoracic duct. The patient was discharged on hospital day eight and was doing quite well at one-year follow-up with no residual effects of his accident. PMID- 11409812 TI - Has nonoperative management of solid visceral injuries adversely affected resident operative experience? AB - The purpose of this study was to assess the impact of increased use of nonoperative management of blunt injuries to the spleen or liver on surgical residents' operative experience with solid visceral injuries. We conducted a 10 year retrospective study of blunt spleen and liver injuries at a state-designated Level I trauma center and a survey of chief residents' operative experience with splenic and hepatic injuries from blunt trauma during the same time period. From 1990 through 1999, 431 patients were admitted with splenic injuries and 634 patients were admitted with liver injuries; 350 splenic injuries (81%) were due to blunt trauma; 317 liver injuries (50%) were caused by blunt mechanisms. In 1990 100 per cent of patients with splenic injuries and 93 per cent of those with liver injuries underwent surgery for those injuries. These rates were 19 and 28 per cent respectively in 1999. The number of patients with blunt solid visceral injuries increased more than fourfold from 1990 through 1999. The number of operations for splenic and hepatic injuries performed by chief residents did not decline significantly during this time period (5.5 cases per chief resident in 1990; 4.6 cases per chief resident in 1999). The increased numbers of patients with solid visceral injuries were due to two factors: increased proportion of blunt trauma admissions especially from motor vehicle collisions and improved recognition of spleen and liver injuries by expanded use of CT scans. We conclude that nonoperative management of blunt solid visceral injuries does not necessarily lead to a diminution of operations nor jeopardize resident education. However, trauma volumes must be high enough to support adequate operative experience. PMID- 11409813 TI - Prospective comparison of intraoperative angiography with duplex scanning in evaluating lower-extremity bypass grafts in a community hospital. AB - The gold standard for intraoperative evaluation of lower-extremity bypass grafts has been angiography. Limitations of this technique include inability to measure flow dynamics, violation of graft integrity, cost, and length of assessment time. The goal of this study was to evaluate duplex scanning as an alternative modality for intraoperative graft assessment. Our study group comprised of 19 consecutive patients undergoing infrainguinal bypass procedures at our institution between March 1999 and March 2000. Intraoperative angiography was compared with duplex scanning by evaluating parameters of assessment time, graft flow velocities, serum creatinine levels, and 30-day graft patency rates. Mean study times were the following: cut-film angiography, 22 +/- 1.8 minutes; real-time fluoroscopy, 17 +/- 2.5 minutes; and duplex imaging, 10.4 +/- 1.1 minutes. As noted duplex imaging times as compared with radiographic modalities were significantly shorter (P < 0.05). There was a substantial cost difference between angiography ($650) and duplex scanning ($350). A 100 per cent correlation of study findings was noted between angiography and duplex scanning. No significant change in pre- versus postoperative creatinine levels was found. We conclude that duplex scanning is an effective modality and provides reliable intraoperative vascular graft assessment data in a community hospital setting. Advantages include a shorter study time, lower cost, flow dynamic data acquisition, and avoidance of mechanical graft trauma. PMID- 11409814 TI - Theodor Billroth: surgeon and musician. PMID- 11409815 TI - The impact of managed care on children's access, satisfaction, use, and quality of care. AB - OBJECTIVE: To examine the impact of managed care on children's access, satisfaction, use, and quality of care using nationally representative household survey data. DATA SOURCE: The 1996 Medical Expenditure Panel Survey (MEPS). STUDY DESIGN: Bivariate and multivariate analyses are used to detect independent effects of managed care on access, satisfaction, utilization, and quality of pediatric health services. DATA COLLECTION/EXTRACTION METHODS: Data were obtained from rounds 1, 2, and 3 of the 1996 MEPS. MEPS collects data on health care use, insurance, access, and satisfaction, along with basic demographic and health status information for a representative sample of the U. S. civilian, noninstitutionalized population. Our sample consists of 5,995 children between the ages of 0 and 17. FINDINGS: Among the 18 outcome indicators examined, the bivariate analysis revealed only three statistically significant differences between children enrolled in managed care and children in traditional health plans: children enrolled in managed care were more likely to receive physician services, more likely to have access to office-based care during evening or weekend hours, and less likely to report being very satisfied with overall quality of care. However, after controlling for confounding factors, none of these differences remained statistically significant. CONCLUSIONS: Our findings suggest that there are no statistically significant differences in self-reported outcomes for children enrolled in managed care and traditional health plans. This conclusion is provisional, however, because of limitations in the data set. PMID- 11409816 TI - The cost and cost-effectiveness of an enhanced intervention for people with substance abuse problems at risk for HIV. AB - OBJECTIVE: To estimate the costs, effectiveness, and cost-effectiveness of prevention interventions for out-of-treatment substance abusers at risk for HIV. This is the first cost-effectiveness study of an AIDS intervention that focuses on drug use as an outcome. STUDY DESIGN: We examined data from the North Carolina Cooperative Agreement site (NC CoOp). All individuals in the study were given the revised NIDA standard intervention and randomly assigned to either a longer, more personalized enhanced intervention or no additional intervention. We estimated the cost of each intervention and, using simple means analysis and multiple regression models, estimated the incremental effectiveness of the enhanced intervention relative to the standard intervention. Finally, we computed cost effectiveness ratios for several drug use outcomes and compared them to a "back of-the-envelope" estimate of the benefit of reducing drug use. PRINCIPAL FINDINGS: The estimated cost of implementing the standard intervention is $187.52, and the additional cost of the enhanced intervention is $124.17. Cost effectiveness ratios range from $35.68 to $139.52 per reduced day of drug use, which are less than an estimate of the benefit per reduced drug day. CONCLUSIONS: The additional cost of implementing the enhanced intervention is relatively small and compares favorably to a rough estimate of the benefits of reduced days of drug use. Thus, the enhanced intervention should be considered an important additional component of an AIDS prevention strategy for out-of-treatment substance abusers. PMID- 11409817 TI - Asthma prevalence, cost, and adherence with expert guidelines on the utilization of health care services and costs in a state Medicaid population. AB - OBJECTIVE: To provide a descriptive analysis of asthma prevalence and costs in a Medicaid population and gauge the degree of adherence with expert guidelines for asthma medication management from the National Asthma Education and Prevention Program. DATA SOURCES: Kentucky Medicaid administrative data for 1996. STUDY DESIGN: A cross-sectional retrospective analysis was used to determine adherence with asthma medication guidelines and utilization of asthma-related health care services and costs. Multivariate logistic regression was used to determine the relationship between nonadherence with the guidelines and utilization of health care services. PRINCIPAL FINDINGS: Of the 530,000 Medicaid recipients, 24,365 (4.6 percent) were identified as having asthma. Average annual asthma-related costs ($616) accounted for less than 20 percent of total health care costs ($3,645). Nonadherence to the guidelines was prevalent. Less than 40 percent of the patients received a prescription for a rescue medication, and fewer than 10 percent of the patients who received daily inhaled short-acting beta-2 agonists were regular users of inhaled steroids. Nonadherence to the guidelines was associated with an increased risk of an asthma-related hospitalization (odds ratio = 1.5, p < .05). CONCLUSIONS: Guideline nonadherence was widespread and associated with an increase in exacerbations of asthma that resulted in hospitalizations. Asthma prevalence and utilization of health care services in a Medicaid population were similar to previous estimates reported nationally and in health maintenance organizations. PMID- 11409818 TI - Changes in prenatal care timing and low birth weight by race and socioeconomic status: implications for the Medicaid expansions for pregnant women. AB - OBJECTIVE: To conduct the first national study that assesses whether the Medicaid expansions for pregnant women, legislated by Congress over a decade ago, met the policy objectives of improved access to care and birth outcomes for poor and near poor women. DATA SOURCES/STUDY SETTING: Data on 8.1 million births using the 1980, 1986, and 1993 National Natality Files. We use births from all areas of the United States except California, Texas, Washington, and upstate New York. METHODS: We conduct a before and after analysis that compares obstetrical outcomes by race and socioeconomic status for the periods 1980-86 and 1986-93. We examine whether women of low socioeconomic status showed greater improvements in outcomes during the 1986-93 period compared to the 1980-86 period. We analyze two obstetrical outcomes: the rate of late initiation of prenatal care and the rate of low birth weight. DATA COLLECTION: Natality data were aggregated to race, socioeconomic status, age, and parity groups. RESULTS: During the 1986-93 period, rates of late initiation of prenatal care decreased by 6.0 to 7.8 percentage points beyond changes estimated for the 1980-86 period for both white and African American women of low socioeconomic status. For some white women of low socioeconomic status, the rate of low birth weight was reduced by 0.26 to 0.37 percentage points between 1986 and 1993 relative to the earlier period. Other white women of low socioeconomic status and all African American women of low socioeconomic status showed no relative improvement in the rate of low birth weight during the 1986-93 period. CONCLUSIONS: The expansions in Medicaid lead to significant improvements in prenatal care utilization among women of low socioeconomic status. The emerging lesson from the Medicaid expansions, however, is that increased access to primary care is not adequate if the goal is to narrow the gap in newborn health between poor and nonpoor populations. PMID- 11409819 TI - Prenatal care--necessary but not sufficient. PMID- 11409820 TI - The role of public clinics in preventable hospitalizations among vulnerable populations. AB - OBJECTIVE: To determine if the availability of public ambulatory clinics affects preventable hospitalization (PH) rates of low-income and elderly populations. DATA SOURCES: PH rates were calculated using elderly and low-income discharges from 1995-97 Virginia hospital discharge data. Other data sources include the 1990 Census, the 1998 Area Resource File, the 1996 American Hospital Association Survey, the Virginia Department of Health, the Virginia Primary Care Association, and the Bureau of Primary Health Care. STUDY DESIGN: Multiple linear regression was used to evaluate the relationship between ambulatory clinic availability and PH rates, controlling for population and other provider characteristics in a cross-section of zip code clusters. DATA EXTRACTION METHODS: Clusters with populations of at least 2,000 were assembled from zip codes in each county in the state of Virginia. Overlapping medical market service areas were constructed around the population centroid of each cluster. PRINCIPAL FINDINGS: Populations in medically underserved areas (MUAs) served by a Federally Qualified Health Center had significantly lower PH rates than did other MUA populations. The presence of a free clinic had a marginally significant association with lower PH rates. CONCLUSIONS: The availability of public ambulatory clinics is associated with better access to primary care among low-income and elderly populations. PMID- 11409821 TI - Strategies for cutting hospital beds: the impact on patient service. AB - OBJECTIVE: To develop insights on the impact of size, average length of stay, variability, and organization of clinical services on the relationship between occupancy rates and delays for beds. DATA SOURCES: The primary data source was Beth Israel Deaconess Medical Center in Boston. Secondary data were obtained from the United Hospital Fund of New York reflecting data from about 150 hospitals. STUDY DESIGN: Data from Beth Israel Deaconess on discharges and length of stay were analyzed and fit into appropriate queueing models to generate tables and graphs illustrating the relationship between the variables mentioned above and the relationship between occupancy levels and delays. In addition, specific issues of current concern to hospital administrators were analyzed, including the impact of consolidation of clinical services and utilizing hospital beds uniformly across seven days a week rather than five. PRINCIPAL FINDINGS: Using target occupancy levels as the primary determinant of bed capacity is inadequate and may lead to excessive delays for beds. Also, attempts to reduce hospital beds by consolidation of different clinical services into single nursing units may be counterproductive. CONCLUSIONS: More sophisticated methodologies are needed to support decisions that involve bed capacity and organization in order to understand the impact on patient service. PMID- 11409822 TI - Putting research to work: reporting and enhancing the impact of health services research. PMID- 11409823 TI - Reading my palm: adventures in medicine. PMID- 11409824 TI - Health status of children with moderate to severe cerebral palsy. AB - The aim of the study was to evaluate the health of children with cerebral palsy (CP) using a global assessment of quality of life, condition-specific measures, and assessments of health care use. A multicenter population-based cross sectional survey of 235 children, aged 2 to 18 years, with moderate to severe impairment, was carried out using Gross Motor Function Classification System (GMFCS) levels III (n = 56), IV (n = 55), and V (n = 122). This study group scored significantly below the mean on the Child Health Questionnaire (CHQ) for Pain, General Health, Physical Functioning, and Impact on Parents. These children used more medications than children without CP from a national sample. Fifty-nine children used feeding tubes. Children in GMFCS level V who used a feeding tube had the lowest estimate of mental age, required the most health care resources, used the most medications, had the most respiratory problems, and had the lowest Global Health scores. Children with the most severe motor disability who have feeding tubes are an especially frail group who require numerous health-related resources and treatments. Also, there is a relationship among measures of health status such as the CHQ, functional abilities, use of resources, and mental age, but each appears to measure different aspects of health and well-being and should be used in combination to reflect children's overall health status. PMID- 11409825 TI - Comparison of three ankle-foot orthosis configurations for children with spastic hemiplegia. AB - The purpose of this study was to examine the effectiveness of the hinged ankle foot orthosis (HAFO), posterior leaf spring (PLS), and solid ankle-foot orthosis (SAFO), in preventing contracture, improving efficiency of gait, and enhancing performance of functional motor skills in 30 children (21 male, 9 female; mean age 9 years 4 months; age range 4 to 18 years,) with spastic hemiplegia. Following a 3-month baseline period of no ankle-foot orthosis (AFO) use, each AFO was worn for 3 months after which ankle range of motion, gait analysis, energy consumption, and functional motor skills were assessed. The HAFO and PLS increased passive ankle dorsiflexion and normalization of ankle rocker function during gait. Normalization of knee motion in stance was dependent upon the knee abnormality present and AFO configuration. The HAFO was the most effective in controlling knee hyperextension in stance, while PLS was the most effective in promoting knee extension in children with >10 degree knee flexion in stance. Energy efficiency was improved in 21 of the children, with 13 of these children demonstrating the greatest improvement in HAFO and PLS. Improvements in functional mobility were greatest in the HAFO and PLS. PMID- 11409826 TI - Management of drooling in individuals with neurodisability: a surgical experience. AB - From 1975 to 1 January 1999, 1103 neurologically involved patients (mean age 13.2 years; 686 males, 417 females) referred with problematic drooling, or sialorrhea, were assessed at a pediatric rehabilitation center by a team consisting of an otolaryngologist, speech pathologist, and a dentist. The initial standard treatment for persistent sialorrhea (in the compliant or aware patient) is oral motor training. A group of 522 patients with persistent significant drooling after a minimum of 6 months of oral-motor training, or profuse drooling in the presence of a low cognitive level, underwent surgery, usually when over 6 years of age. From 1978 to 1 January 1998, the operation of first choice was submandibular duct relocation (SDR), and was completed in a total of 226 patients. Midway through 1988, sublingual gland excision was also completed at the time of submandibular duct relocation (SDRSGE); 249 of these procedures have been completed to January 1st 1999. Those patients who had SDRSGE had significantly fewer complications that required additional surgery than those that had SDR only. However, the impact of surgery on the drooling as evaluated in subsets of both groups (SDR n=115, SDRSGE n=106) was statistically similar. The study of 11 children with salivary gland radionuclitide scans to determine the effect of submandibular duct surgery on gland function was inconclusive. The pattern of oral-motor function in 26 children studied after SDRSGE surgery suggested that those children with severe impairment of volitional motor function and profuse drooling tended to have a poorer outcome following surgery compared to those with milder impairments. PMID- 11409827 TI - Assessment of upper-limb function and movement in children with cerebral palsy wearing lycra garments. AB - It has recently been suggested that lycra garments are helpful for children with cerebral palsy (CP). Twelve children, with athetosis, ataxia, and spasticity, were fitted with lycra garments (Kendall-Camp UK Ltd). Scores on the Paediatric Evaluation of Disability Inventory (PEDI) scales were determined before and after wearing the garment for at least 6 hours a day for 6 weeks. Five children with motor problems representative of the whole group were investigated during a reach and-grasp task by kinematic motion analysis; reflective markers were used with and without the garment. Carers were given a questionnaire concerning the practicalities of using the garments. All 12 children made improvements in at least one of the functional scales of the PEDI, and scores for the whole group showed significant gains (Wilcoxon chi2 test, self-help p<0.01; mobility p<0.5; social p<0.1). These changes were usually slight, although noticed by carers. Six children made gains of at least one scale of the caregiver assistance scores, two of the children showed losses (due to difficulties removing the garment for toileting), and four showed no change. Motion analysis indicated that (1) two children with athetosis had improved proximal stability in sitting and in smoothness of arm movements, (2) one child with ataxia had improved in proximal and distal stability, and (3) two children with spasticity had more jerky movements, although one improved in proximal stability. All children had problems in wearing the garments, including problems with toileting and incontinence of urine; the parents of only one child wanted to continue using it. Results suggest that the functional benefit of lycra garments for children with CP is mainly due to improvements in proximal stability but this should be weighed against the inconvenience and loss of independence. PMID- 11409828 TI - Clinical findings, consanguinity, and pedigrees in children with anophthalmos in southern India. AB - This study aimed to describe clinical findings, pedigrees, and possible environmental risk factors in children with clinical anophthalmos and remnant microphthalmos in either eye in southern India. Twenty-four children (14 male, 10 female; mean age 10.3 years, age range 1.3 to 18 years,) were recruited from schools for the blind, hospitals, and community-based rehabilitation programmes in Andhra Pradesh, India, over 1 year. Family members were examined, and mothers interviewed. Fifteen children had anophthalmos and nine had remnant microphthalmos in one or both eyes. Twelve children had associated systemic findings, of which six were major and six were minor abnormalities. Information on consanguinity was available in 19 children, 12 of whom had consanguineous parents. Five children had a positive family history. Two mothers had a history of night blindness, and one had a history of pesticide exposure during pregnancy. High rates of consanguinity suggest a genetic recessive aetiology. PMID- 11409829 TI - Sensory modulation dysfunction in children with attention-deficit-hyperactivity disorder. AB - This study investigates the presence of sensory modulation dysfunction (SMD) among children with attention-deficit-hyperactivity disorder (ADHD). Twenty-six children with ADHD (mean age 8.3 years, 18 males, 8 females), and 30 typically developing children (mean age 8.2 years, 21 males, 9 females) were tested using a laboratory procedure that gauges responses to repeated sensory stimulation by measuring electrodermal reactivity (EDR). Parental report measures of limitations in sensory, emotional, and attentional dimensions were administered using the Short Sensory Profile, the Leiter International Performance Scale-Revised, Parent Rating subscales, and the Child Behavior Checklist (CBCL). Compared to the typical sample, the children with ADHD displayed greater abnormalities in sensory modulation on both physiological and parent-report measures. The children with ADHD also displayed more variability in responses. Within the group with ADHD, levels of SMD were highly correlated with measures of psychopathology on the CBCL. Implications of findings relate to the importance of considering sensory processing abilities in a subgroup of children with ADHD. PMID- 11409830 TI - Neuropsychological disorders related to interictal epileptic discharges during sleep in benign epilepsy of childhood with centrotemporal or Rolandic spikes. AB - Nine children (five males, four females; age range 6 years 1 month to 11 years 1 month) affected by benign epilepsy of childhood with centrotemporal or Rolandic spikes (BECRS) with EEG evidence of marked activation of interictal epileptic discharges (IEDs) during sleep, and nine unaffected control children matched for age, sex, and socioeconomic status, were enrolled in a prospective study. At the time of detection of IED activation during sleep, patients showed a mean Full Scale IQ score within the normal range, but significantly below that of control participants; neuropsychological assessment revealed disorders in visuospatial short-term memory (Corsi's Block Tapping Test), attention, and cognitive flexibility (Trail Making Test and Stroop Color-Word Test), picture naming, and fluency (Benton's Naming Test and Word Fluency), visuoperceptual skill (Ghent Poppelreuter and Street Gestalt Completion Tests) and visuomotor coordination (Bender Test). After detection of IED activation during sleep, children were followed up for 2 years. At the time of IED remission (T1), neuropsychological re evaluation showed a notable increase in IQ score and a significant improvement (t test: p<0.007) in visuomotor coordination, non-verbal short-term memory, sustained attention and mental flexibility, picture naming, and visual-perceptual performance. At T1, patients' performance did not differ from the controls (Mann Whitney U test). PMID- 11409831 TI - Pyridoxine-dependent seizures responding to extremely low-dose pyridoxine. AB - We report on a male infant with pyridoxine dependency and seizures from birth, controlled with pharmacological doses of pyridoxine at 4 months of age. Seizures stopped between 30 and 80 days of age when very-low doses of pyridoxine were given in a multivitamin supplement. Daily dose was 0.5 mg that corresponded to 0.08 to 0.16 mg/kg/day when weight gain is considered. In previous reports doses have ranged from 0.2 to 30 mg/kg/day. Another distinctive feature was that this infant went into a coma and developed hypotonia and irregular breathing when pyridoxine was given by enteral tube which has usually been reported when the vitamin is given intravenously. Use of low doses of pyridoxine in multivitamin supplements could be a confounding factor for early diagnosis and appropriate treatment of pyridoxine-dependent seizures. PMID- 11409832 TI - Pyridoxine-dependent and pyridoxine-responsive seizures. PMID- 11409833 TI - Facial electromyography in newborn and young infants with congenital facial weakness. PMID- 11409834 TI - 'An international perspective on Tourette syndrome'. PMID- 11409835 TI - Non-invasive technique for assessment and management planning of oral-pharyngeal dysphagia in children with cerebral palsy. PMID- 11409836 TI - HHH syndrome associated with callosal agenesis and disordered neuronal migration. PMID- 11409837 TI - Heart--the source and target of "hypothalamic neurohormones": do they provide a delicate regulation complementary to the conduction system in the heart? PMID- 11409838 TI - Effects of three alkoxypsoralens on voltage gated ion channels in Ranvier nodes. AB - The effects of the phototoxic K+- channel blockers 8-methoxypsoralen (8-MOP) and 5-methoxypsoralen (5-MOP) on Ranvier nodes were compared to those of 5,8 diethoxypsoralen (5,8-EOP) by means of the Hodgkin-Huxley formalism. When these test substances were added individually to the bathing solution (8-MOP: 100 micromol/l; 5-MOP: 50 micromol/l; 5,8-EOP: 10 micromol/l) the following completely reversible effects were observed: 1. 8-MOP, caused a nearly potential independent decrease of the sodium permeability, P'Na, by ca. 17%. 5-MOP and 5,8 EOP merely decreased the maximal value of P'Na, by ca. 12 and 8% respectively, whereas with weak depolarisations P'Na was unchanged. 2. In the tested potential range the potassium permeability, P'K, was caused to decrease by ca. 9% by 8-MOP, ca. 21% by 5-MOP and ca. 19% by 5,8-EOP. 3. The potassium currents acquired a phasic time course previously described for 8-MOP and 5-MOP. They reached a relative maximum and approached a lower steady-state value, kinfinity, with a time constant tauk at V = 120 mV of about 16 ms (8-MOP), 20 ms (5-MOP) and 94 ms (5,8-EOP). To obtain dose-response relations the drug-induced effects on peak P'K and on the steady state value, kinfinity, were measured. The corresponding apparent dissociation constants (in micromol/l) were 66.6 and 80.1 (for 8-MOP), 87.6 and 25.8 (for 5-MOP), and 13.5 and 6.5 (for 5,8-EOP). In view of the similarity of the actions of 5-MOP and 5,8-EOP as well as the fact that 5,8-EOP is not phototoxic, in future 5,8-EOP may well prove to be a particularly suitable K+-channel blocker for the symptomatic therapy of multiple sclerosis and other demyelinating diseases. PMID- 11409839 TI - Antihemolytic effect of Rooibos tea (Aspalathus linearis) on red blood cells of Japanese quails. AB - The antihemolytic activity of Rooibos and black tea on Japanese quail erythrocytes was studied. Peroxide and hypotonic hemolysis of the red blood cells of quails, either fed with Rooibos tea supplemented food or fed without tea, was performed. Long-term consumption of Rooibos tea did not change the erythrocyte fragility to either peroxide or hypotonia induced hemolysis. However, Rooibos and black teas decreased peroxide induced hemolysis of erythrocytes incubated with each of them, but not hemolysis induced by hypotonic NaCl solution. Stronger inhibition of hemolysis has been obtained when a boiled water extract of Rooibos tea was used for the inhibition. The degree of inhibition was comparable with the effect of ascorbic acid. PMID- 11409840 TI - Synergistic effect of vitamin C on DNA damage induced by cadmium. AB - Salts of divalent cadmium are well-known human mutagens and carcinogens. In the present work, the ability of vitamin C to modulate genotoxic effects of cadmium chloride on human lymphocytes was assessed using single cell gel electrophoresis (comet assay). Vitamin C at 20 and 100 micromol/l and cadmium at 5, 30 and 150 micromol/l significantly increased the tail moment of lymphocytes. Vitamin C also increased the tail moment of cells exposed to cadmium. This effect was concentration-dependent: the higher the vitamin C concentration the greater the tail moment. The combined effects of cadmium and vitamin C were more pronounced at all concentrations tested than the sum of the effects of the compounds applied separately (p < 0.05), so cadmium and vitamin C can be considered to have synergistic effects. The results obtained can be partly explained by the participation of cadmium in the Fenton reaction and reduction of its oxidized form by vitamin C. PMID- 11409841 TI - Decreased fluidity of isolated erythrocyte membranes in type 1 and type 2 diabetes. The effect of resorcylidene aminoguanidine. AB - Changes in the physico-chemical properties of erythrocyte membranes induced by nonenzymatic glycation as well as the possible prevention of their rise were studied. Using the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH), fluorescence anisotropy values were determined in erythrocyte membranes isolated from type 1 and type 2 diabetic patients with and without complications. The mean anisotropy values for the groups of diabetic patients were significantly higher than those for the control group (p < 0.01). This indicated pathologically decreased fluidity in cell membranes in the diabetics regardless of the type of diabetes or the presence of complications. The fluorescence anisotropy positively correlated (p < 0.01) with clinical parameters, such as glycohaemoglobin and plasma cholesterol content, which are important for the monitoring of the compensation status of the diabetic patient. Our results support the suggestion that protein crosslinking and oxidative stress induced by nonenzymatic glycation contribute to changes in the physico-chemical properties of erythrocyte membranes. In vitro testing of a new potential drug resorcylidene aminoguanidine (RAG) showed its ability to increase significantly (p < 0.001), to various extent (p < 0.01), the fluidity of both diabetic and control erythrocyte membranes. Upon the administration of RAG, reduced fluorescence anisotropy values for the groups of diabetic patients approached the normal values obtained for the controls. This may play an important role in the improvement of impaired cell functions found in diabetes that are controlled by the cell membrane. PMID- 11409842 TI - Human blood platelets, PMN leukocytes and their interactions in vitro. Responses to selective and non-selective stimuli. AB - Using simultaneous recording of aggregation and chemiluminescence, responses of human polymorphonuclear leukocytes, blood platelets and their mixture were investigated after stimulation by specific as well as non-specific stimuli for each cell. In our experimental settings, aggregation of platelets and PMN leukocytes was increased in the following order of stimuli: PMA 0.05) elevation in the content of conjugated dienes as well as by a new wave of depression of the SOD activity (p < 0.05) in both the CE and the ThL segment. From our results it could be concluded that all considerable damage to the spinal cord occurred during the ischemic period. In the period of reperfusion reparative changes started to predominate. This is in accordance with the recent discoveries indicating that, when coupled with an increase in tissue gamma-GTP activity, the post-ischemic reparative changes comprise a replenishment of the cell glutathione pool. This process is accompanied with a gradual increase in H2O2 production that results in repeatead inhibition of the SOD activity and a tendency to conjugated dienes formation. PMID- 11409845 TI - Ping-pong character of nasturtium-seed xyloglucan endotransglycosylase (XET) reaction. AB - Plant xyloglucan endotransglycosylase (XET, EC 2.4.1.207) degrades its substrate by a transglycosylation mechanism while endo-cleaving xyloglucan (XG) molecules at their beta-1,4-linked polyglucosyl main chain and transferring the newly generated reducing chain ends to hydroxyls at C-4 of non-reducing glucosyl ends of the main chains of other XG molecules or of low-Mr XG-fragments (OS). Kinetic data obtained with purified nasturtium seed (Tropaeolum majus, L.) XET while using high-Mr xyloglucan and 3H-labeled XGOS alditols (DP 7-9) as substrates could be best fitted to the model for Ping-Pong Bi Bi reaction mechanism. Such mechanism is typical for transglycosylases operating with retention of the anomeric configuration of the formed glycosidic bond and involving the formation of a covalent glycosyl-enzyme reaction intermediate. PMID- 11409846 TI - Dephosphorylation agents depress gap junctional communication between rat cardiac cells without modifying the Connexin43 phosphorylation degree. AB - The functional state of gap junctional channels and the phosphorylation status of Connexine43 (Cx43), the major gap junctional protein in rat heart, were evaluated in primary cultures of neonatal rat cardiomyocytes. H7, able to inhibit a range of serine/threonine protein kinases, progressively reduced gap junctional conductance to approximately 13% of its initial value within 10 min except when protein phosphatase inhibitors were also present. The dephosphorylating agent 2,3 Butanedione monoxime (BDM) produced both a quick and reversible interruption of cell-to-cell communication as well as a parallel slow inhibition of junctional currents. The introduction of a non-hydrolysable ATP analogue (ATPgammaS) in the cytosol delayed the second component, suggesting that it was the consequence of protein dephosphorylation. Western blot analysis reveals 2 forms of Cx43 with different electrophoretic mobilities which correspond to its known phosphorylated and dephosphorylated forms. After exposure of the cells to H7 (1 mmol/l, 1h) or BDM (15 mmol/l, 15 min), no modification in the level of Cx43 phosphorylation was observed. The lack of direct correlation between the inhibition of cell-to-cell communication and changes in the phosphorylation status of Cx43 suggest that the functional state of junctional channels might rather be determined by regulatory proteins associated to Cx43. PMID- 11409847 TI - The integrin-mediated cyclic strain-induced signaling pathway in vascular endothelial cells. AB - The irregular distribution of plaque in the vasculature results from the interaction of local hemodynamic forces with the vessel wall. One well characterized force is cyclic circumferential strain, the repetitive pulsatile pressure distention on the arterial wall. This review summarizes current research, which has aimed to elicit the signal transduction pathway by which cyclic strain elicits functional and structural responses in endothelial cells; specifically, it summarizes the signaling pathway that begins with the reorganization of integrins. One method by which these extracellular matrix receptors affect signal transduction is through their ability to initiate the process of phosphorylation on tyrosine residues of cytoplasmic protein kinases, including focal adhesion kinase. The strain-induced pathway appears to also involve ras and the mitogen-activated protein kinase family of enzymes, and preliminary data suggests a role for src as well. Ultimately, it is the regulation of gene expression through the modulation of transcription factors that allows endothelial cells to respond to changes in local hemodynamics. PMID- 11409848 TI - Vascular endothelial growth factor in porcine-derived extracellular matrix. AB - An extracellular matrix (ECM) derived from the submucosa of the porcine small intestine (SIS) has been shown to induce angiogenesis and host tissue remodeling when used as a xenogeneic bioscaffold in animal models of wound repair. In the present study, we compared the in vitro effects of SIS ECM extracts to several purified angiogenic growth factors on human dermal microvascular endothelial cell (HMEC) growth patterns. The SIS ECM was shown to induce tube formation from HMEC in a three-dimensional fibrin-based angiogenesis assay in a manner similar to that caused by the addition of vascular endothelial growth factor (VEGF). This tube formation was blocked in the presence of anti-VEGF neutralizing antibody. Western blots and ELISA procedures showed that the SIS ECM contains as much as 0.77 ng VEGF/g SIS. The closely related endothelial cell mitogen, platelet derived growth factor (PDGF), was not detectable in the SIS extracts. We conclude that VEGF is present in the SIS extracellular matrix. The role of VEGF in SIS induced wound repair remains unknown, but its presence in the ECM makes it a possible contributor to the angiogenic effect of SIS when this ECM is used as a tissue repair scaffold in animal models of wound repair. PMID- 11409849 TI - Unique sensitivities to cytokine regulated expression of adhesion molecules in human heart-derived endothelial cells. AB - The expression of adhesion molecules by endothelial cells is crucial in many inflammatory processes and plays an active role in the development of reperfusion injury, acute and chronic rejection. The expression of adhesion molecules in different parts of the coronary tree to cytokine stimulation is not known. We describe here a detailed study of the effects of the inflammatory cytokines TNFalpha and IL-1beta on the expression of adhesion molecules vascular cell adhesion molecule-1 (VCAM-1), E-selectin and intracellular cell adhesion molecule 1 (ICAM-1) on human aortic root (HAEC), coronary artery (HCAEC) and heart microvascular (HHMEC)) endothelial cells in culture, using flow cytometry. We found constitutive levels of both VCAM-1 and E-Selectin on HCAEC and HHMEC (approximately 20%) which were significantly higher compared to HAEC (approximately 3%). There was an extreme sensitivity of HCAEC and HHMEC to 0.002 ng/ml TNFalpha: (VCAM-1 approximately 40%, E-Selectin approximately 25%) respectively, compared to HAEC (VCAM-1 approximately 5%, E-selectin approximately 5%). IL-1beta showed a similar pattern of expression at low doses (5 U/ml), but was less potent. We also observed prolonged expression of these adhesion molecules, especially on the HHMEC (>48 hours) compared to HAEC. There was also increased binding of peripheral blood mononuclear cells (PBMC) to both non stimulated and TNFalpha stimulated HCAEC and HHMEC compared to HAEC. This data suggest that endothelial cells in different regions of the coronary tree express different patterns of basal and cytokine-stimulated adhesion molecule expression. PMID- 11409850 TI - Cyclic stretch induces the expression of vascular endothelial growth factor in vascular smooth muscle cells. AB - OBJECTIVE: Accumulating evidence links the release of vascular endothelial growth factor (VEGF) by vascular smooth muscle cells (VSMC) to normal endothelial cell (EC) function, repair and maintenance. Using an in vitro model we investigate the role of cyclic stretch on both the release of VEGF by VSMC and the phosphorylation of a VEGF receptor on EC. METHODS: Bovine VSMC and EC were exposed to 10% cyclic strain for 4 hours. VEGF mRNA steady-state levels of VSMC were analysed by northern blot hybridisation. The presence of secreted VEGF from VSMC was determined by assaying the migration of EC. VEGF receptor phosphorylation on stretched EC was assayed by immunoblotting. RESULTS: The steady-state level of VEGF mRNA in stretched VSMC increased 3.3 (+/- 0.6) fold above that of unstretched VSMC (p < 0.005). Migration of EC was stimulated 8.3 (+/- 1.1) and 14.6 (+/- 1.3) fold by media from unstretched and stretched VSMC respectively, demonstrating a 1.8 fold increase due to stretch alone (p < 0.05). Cyclic stretch resulted in phosphorylation of the VEGF receptor KDR. CONCLUSION: Exposure of VSMC to physiological levels of stretch induces a biologically significant increase in VEGF secretion and may provide an arterial stimulus for maintenance of steady state levels of VEGF essential for EC survival. PMID- 11409851 TI - Expression of VACM-1 protein in cultured rat adrenal endothelial cells is linked to the cell cycle. AB - The vasopressin-activated calcium-mobilizing (VACM-1) protein is a unique arginine vasopressin (AVP) receptor which shares sequence homology with the cullins, genes involved in the regulation of cell cycle transitions. Unlike either cullins or AVP receptors, however, VACM-1 is expressed exclusively in the vascular endothelial cells and in the renal collecting tubule cells. In order to test the hypothesis that the expression of VACM-1 might be correlated with the cell cycle, and to establish an endothelial cell model for the VACM-1 receptor, we examined VACM-1 expression in rat adrenal medulla endothelial cells (RAMEC). Northern and Western blot analyses of mRNA and protein from RAMEC identified presence of 6.4 kb mRNA and a Mr 81 kDa protein, respectively. Immunostaining of RAMEC with anti-VACM-1 antibodies and Western blot analyses indicated that in RAMEC, VACM-1 protein expression is dependent on the cell cycle. VACM-1 protein virtually disappears during the S phase and localizes to the cytosol during cell division and to the cell membrane at the completion of cytokinesis. Furthermore, pretreatment of RAMEC with anti-VACM-1 specific antibodies increased basal levels of Ca2+and attenuated the AVP-dependent increase in cytosolic Ca2+. In summary, these results indicate that VACM-1 protein expression in RAMEC membrane is linked to the cell cycle, and consequently, VACM-1 may be involved in the regulation of cell division. PMID- 11409852 TI - Activation of endothelial cell mitogen activated protein kinase ERK(1/2) by extracellular HIV-1 Tat protein. AB - Extracellular Tat protein, the transactivating factor of the human immunodeficiency virus type 1 (HIV-1), modulates gene expression, growth, and angiogenic activity in endothelial cells by interacting with the vascular endothelial growth factor (VEGF) receptor-2 (Flk-1/KDR). Recombinant Tat protein, produced as glutathione-S-transferase chimera (GST-Tat), activates mitogen activated protein kinase (MAPK) ERK(1/2) in human, murine, and bovine endothelial cells whereas GST is ineffective. In bovine aortic endothelial cells, GST-Tat and the 165 amino acid VEGF isoform (VEGF165) induce transient ERK(1/2) phosphorylation with similar potency and kinetics. The synthetic peptide Tat(41 60), but not peptides Tat(1-21) and Tat(71-86), causes ERK(1/2) phosphorylation, thus implicating Tat/KDR interaction in the activation of this signalling pathway. Accordingly, GST-Tat induces ERK(1/2) phosphorylation in KDR-transfected porcine aortic endothelial cells but not in parental cells. MAPK kinase inhibitors PD098059 and U0126 prevent ERK(1/2) phosphorylation by Tat. However, they do not affect the angiogenic activity exerted by Tat in the murine Matrigel plug and chick embryo chorioallantoic membrane assays. Blocking of MAPK kinase activity impairs instead the angiogenic response to VEGF165 and to fibroblast growth factor-2 (FGF-2). Our data demonstrate that ERK(1/2) activation following the interaction of HIV-1 Tat protein with endothelial cell Flk-1/KDR receptor does not represent an absolute requirement for a full angiogenic response to this growth factor that appears to utilize mechanism(s) at least in part distinct from those triggered by other prototypic angiogenic growth factors. PMID- 11409853 TI - Contralateral suppression of transient evoked otoacoustic emissions in patients with cerebello-pontine angle tumor. AB - OBJECTIVE: During measurement of transient evoked otoacoustic emissions (TEOAEs), acoustic stimulation of the contralateral ear reduces or suppresses TEOAE amplitude. This is thought to be due to the inhibitory control that the medial efferent auditory nerve exerts on outer hair cell (OHC) function. The main aim of this study was to investigate the effect of cerebello-pontine angle (CPA) tumor on the medial efferent nerve pathways to both tumor and non-tumor ears by examining alterations in TEOAE amplitude that result from contralateral acoustic stimulation. DESIGN: Contralateral suppression of TEOAEs using broadband noise was measured preoperatively in 17 patients with unilateral CPA tumor and 17 normally hearing controls, matched for age and gender. RESULTS: The control ears demonstrated significantly more suppression than the tumor and non-tumor ears in the patient group. There was, however, no significant difference in suppression between the tumor and non-tumor ears, and the statistical correlation for suppression between them was high. There was no effect of gender, hearing threshold levels, or size and type of tumor on suppression, although there was an effect of age on suppression in both the control and patient groups where suppression reduced as age increased. Four of the 17 patients had TEOAEs, which were clearly present in the tumor ear despite substantial hearing loss, three of which had no measurable hearing. CONCLUSIONS: It is hypothesized that neural compression by CPA tumor disrupts the medial efferent nerve control mechanism to the OHCs of tumor ears. It also is hypothesized that neural compression reduces transmission of afferent nerve impulses from the tumor ear, which cross over to the medial olivo-cochlear complex and reduce the inhibitory control of OHC function in the non-tumor cochlea. PMID- 11409854 TI - "Linear" and "derived" otoacoustic emissions in newborns: a comparative study. AB - OBJECTIVE: To investigate the effects of a specific aspect of the acquisition procedure, the averaging technique, on the evaluation of click-evoked otoacoustic emissions (CEOAEs) in newborns. DESIGN: CEOAEs were recorded by an Otodynamic ILO88 system from 89 full-term newborns at the third day after delivery. For each ear and in the same test session, CEOAEs were evoked by 75 to 85 dB pSPL acoustic clicks and averaged according to two different modes: the "linear" (classic average) and the "derived" mode, which allows the cancellation of linear behaving components (such as acoustic artifacts). All examined ears had a normal auditory function as assessed by conventional ABR between the ages of 2 and 4 mo. CEOAEs obtained by both averaging techniques were compared on the basis of several quantitative parameters: the waveform similarity; the levels of signal and noise and the inter-test reproducibility of the broadband response and of four different frequency bands centered at 1.6, 2.4, 3.2, and 4 kHz; the amplitude as a function of time; the test time. To eliminate the contribution of the stimulus artifact, linear CEOAEs were windowed 6 to 20 msec, whereas derived emissions were windowed using the default ILO88 window (2.5 to 20 msec). Additionally, CEOAEs were classified as "pass" or "fail" accordingly to screening criteria used in the daily clinical practice. RESULTS: Linear and derived emissions had very similar wave shapes and no time shifts during the first 12.5 msec. On the contrary, clear differences in the waveforms and time shifts were observed at longer latencies. The use of both averaging techniques resulted in identical CEOAE levels for both the broadband response and for the first two tested frequencies. For the last two frequencies, emission levels were lower when averaged with the linear technique owing to the use of the time window 6 to 20 msec, which reduces the amplitudes of high-frequency components. The residual noise in derived traces is 6 dB higher than that from linear traces. Also, derived CEOAEs had a lower inter-test reproducibility in both the broadband compound emission and in the four frequency bands examined here. The greatest difference in reproducibility was observed at the lowest band (1.2 to 2 kHz). Scoring of emissions was influenced by the averaging technique: 14% CEOAEs obtained with linear averaging and scored as passes were classified as fails when averaged with the derived mode. Moreover, if a CEOAE was scored as pass when using the derived technique, it also was scored as pass when using linear averaging. The increased number of false positives most likely was due to the higher noise floor/lower signal to noise ratio (SNR) of CEOAEs obtained with the derived technique. CONCLUSIONS: In the tested newborns and at the levels of stimulation used in this study, the emissions obtained with the derived technique were noisier than those obtained with the linear technique, this being intrinsically due to the type of averaging. Therefore, screening criteria based on the evaluation of the SNR (or similar parameters) could be influenced by the type of averaging used during the acquisition. PMID- 11409855 TI - The effect of short-term ventilation tubes versus watchful waiting on hearing in young children with persistent otitis media with effusion: a randomized trial. AB - OBJECTIVE: To study the effect of short-term ventilation tubes in children aged 1 to 2 yr with screening-detected, bilateral otitis media with effusion (OME) persisting for 4 to 6 mo, as compared with watchful waiting. DESIGN: Multi-center randomized controlled trial (N = 187) with two treatment arms: short-term ventilation tubes versus watchful waiting. Young children underwent auditory screening; those with persistent (4 to 6 mo) bilateral OME were recruited. RESULTS: The mean duration of effusion over 1-yr follow-up was 142 days (36%) in the ventilation tube (VT) group versus 277 days (70%) in the watchful waiting (WW) group. After 6 mo of follow-up, the pure-tone average in the VT group was 5.6 dB A better than that in the WW group. After 12 mo, most of the advantage in the VT group had disappeared. After the insertion of ventilation tubes, the children with poorer hearing levels at randomization improved more than the children with better hearing levels. The largest difference in hearing levels was found between the children in the VT group whose ventilation tubes remained in situ and the children in the WW group. In the VT children with recurrence of OME, the hearing levels again increased, but remained slightly lower than those in the infants with persistent OME in the WW group. CONCLUSIONS: Ventilation tubes have a beneficial effect on hearing in the short run (6 mo); this effect, however, largely disappears in the long run (12 mo). This is probably due to partial recurrent OME in the VT group and to partial spontaneous recovery in the WW group. PMID- 11409856 TI - Steady state responses to multiple amplitude-modulated tones: an optimized method to test frequency-specific thresholds in hearing-impaired children and normal hearing subjects. AB - OBJECTIVE: To evaluate, using statistical methods, the usefulness of the binaural multiple frequency auditory steady state responses (MF SSRs) for objective, frequency-specific audiometry in a large sample of hearing-impaired children and normal-hearing subjects. DESIGN: The MF SSRs were recorded in a sample of 43 hearing-impaired children (86 ears) and 40 normal-hearing young adults (80 ears). Simultaneous carrier tones (0.5, 1, 2, and 4 kHz) modulated in amplitude at different rates (77 to 105 Hz) were presented binaurally (TDH 49 earphones) at variable intensities (110 to 20 dB SPL). For each subject the response thresholds (RTHs) at 0.5, 1, 2, and 4 kHz, were determined automatically (F test) and compared with the corresponding behavioral thresholds (BTHs). RESULTS: In the normal-hearing subjects, RTHs were detectable, on average, between 11 and 15 dB above the BTH. These differences were significantly smaller in the hearing impaired (5 to 13 dB). Also a close correspondence was found between the subjective and objective audiogram curves in both groups. The within subject Spearman correlation coefficients calculated between the two curves, were in most cases above the significance cut off point (p < 0.05). Also in 1-way repeated measures analysis of variance, the overall error in the estimation of the audiogram (vector across frequency of absolute distances between the curves) did not differ significantly from zero. CONCLUSIONS: The binaural MF SSR was proven to be a valid technique for the estimation of an objective audiogram, in a large sample of hearing-impaired children and normal-hearing subjects. With this method, frequency-specific thresholds at 0.5, 1, 2, and 4 kHz could be determined in all subjects (and both ears) with no appreciable loss in accuracy and a considerable reduction in testing time (average recording time = 21 minutes) when compared with other frequency-specific techniques. PMID- 11409857 TI - Maximizing effective audibility in hearing aid fitting. AB - OBJECTIVE: This paper examines why more audibility is not always better than less audibility if hearing-impaired people are to best understand speech. DESIGN: We used speech perception data from 14 normally hearing and 40 hearing-impaired people to quantify the contribution of audibility to speech intelligibility. The quantification revealed that the effectiveness of audibility decreased with hearing loss, and the decrement was greater at high frequencies than at lower frequencies. To apply the Speech Intelligibility Index (SII) model to predict speech intelligibility for hearing-impaired people, we modified the model to take account of effective audibility rather than physical audibility. RESULTS: The modified SII model provided an adequate description of speech performance of people with a wide range of hearing threshold levels. We applied the model to the evaluation of two prescriptions for a sloping audiogram at prescribed levels and at equated loudness levels to demonstrate the necessity of considering loudness and effective audibility in prescribing amplification. Effective audibility is defined as audibility corrected for the effects of level distortion and hearing loss desensitization, and this paper proposes a method of estimating effective audibility from hearing threshold level at different frequencies. CONCLUSIONS: The practical implication of considering effective audibility in prescribing hearing aids is that for a given listening level, less gain is provided at frequencies where the hearing is most impaired to allow more gain at frequencies where audibility is most useful. In developing the NAL-NL1 prescription for nonlinear hearing aids, we adopted the modified SII model together with a loudness model to derive optimal gain-frequency response characteristics that maximize predicted speech intelligibility for people with different degrees of hearing losses. PMID- 11409858 TI - A single-band envelope cue as a supplement to speechreading of segmentals: a comparison of auditory versus tactual presentation. AB - OBJECTIVE: The objective of this study was to compare the effects of a single band envelope cue as a supplement to speechreading of segmentals and sentences when presented through either the auditory or tactual modality. DESIGN: The supplementary signal, which consisted of a 200-Hz carrier amplitude-modulated by the envelope of an octave band of speech centered at 500 Hz, was presented through a high-performance single-channel vibrator for tactual stimulation or through headphones for auditory stimulation. Normal-hearing subjects were trained and tested on the identification of a set of 16 medial vowels in /b/-V-/d/ context and a set of 24 initial consonants in C-/a/-C context under five conditions: speechreading alone (S), auditory supplement alone (A), tactual supplement alone (T), speechreading combined with the auditory supplement (S+A), and speechreading combined with the tactual supplement (S+T). Performance on various speech features was examined to determine the contribution of different features toward improvements under the aided conditions for each modality. Performance on the combined conditions (S+A and S+T) was compared with predictions generated from a quantitative model of multi-modal performance. To explore the relationship between benefits for segmentals and for connected speech within the same subjects, sentence reception was also examined for the three conditions of S, S+A, and S+T. RESULTS: For segmentals, performance generally followed the pattern of T < A < S < S+T < S+A. Significant improvements to speechreading were observed with both the tactual and auditory supplements for consonants (10 and 23 percentage-point improvements, respectively), but only with the auditory supplement for vowels (a 10 percentage-point improvement). The results of the feature analyses indicated that improvements to speechreading arose primarily from improved performance on the features low and tense for vowels and on the features voicing, nasality, and plosion for consonants. These improvements were greater for auditory relative to tactual presentation. When predicted percent-correct scores for the multi-modal conditions were compared with observed scores, the predicted values always exceeded observed values and the predictions were somewhat more accurate for the S+A than for the S+T conditions. For sentences, significant improvements to speechreading were observed with both the auditory and tactual supplements for high-context materials but again only with the auditory supplement for low-context materials. The tactual supplement provided a relative gain to speechreading of roughly 25% for all materials except low-context sentences (where gain was only 10%), whereas the auditory supplement provided relative gains of roughly 50% (for vowels, consonants, and low-context sentences) to 75% (for high-context sentences). CONCLUSIONS: The envelope cue provides a significant benefit to the speechreading of consonant segments when presented through either the auditory or tactual modality and of vowel segments through audition only. These benefits were found to be related to the reception of the same types of features under both modalities (voicing, manner, and plosion for consonants and low and tense for vowels); however, benefits were larger for auditory compared with tactual presentation. The benefits observed for segmentals appear to carry over into benefits for sentence reception under both modalities. PMID- 11409860 TI - The influence of contextual information on the perception of speech by postlingually and prelingually profoundly hearing-impaired Hebrew-speaking adolescents and adults. AB - OBJECTIVE: To study linguistic context effects on speech perception by prelingually (N = 10) and postlingually (N = 10) hearing-impaired Hebrew-speaking adolescents and adults. DESIGN: Participants were cochlear implant surgery candidates with profound sensorineural hearing loss (95 dB HL+). Four levels of speech perception materials were presented through the auditory-visual modality: monosyllabic nonsense syllables (phonological content), monosyllabic meaningful words (semantic-lexical), sentences without presented topics (semantic syntactic), and sentences with topics (topical). Data were analyzed via percentage correct recognition and via k andj factors (Boothroyd & Nittrouer, 1988). RESULTS: Both participant groups performed better when speech material having more contextual information was presented, with no significant intergroup differences. CONCLUSIONS: This similar usage of contextual information among prelingually and postlingually hearing-impaired participants is encouraging and suggests the role of factors other than age of onset. PMID- 11409859 TI - Use of audiovisual information in speech perception by prelingually deaf children with cochlear implants: a first report. AB - OBJECTIVE: Although there has been a great deal of recent empirical work and new theoretical interest in audiovisual speech perception in both normal-hearing and hearing-impaired adults, relatively little is known about the development of these abilities and skills in deaf children with cochlear implants. This study examined how prelingually deafened children combine visual information available in the talker's face with auditory speech cues provided by their cochlear implants to enhance spoken language comprehension. DESIGN: Twenty-seven hearing impaired children who use cochlear implants identified spoken sentences presented under auditory-alone and audiovisual conditions. Five additional measures of spoken word recognition performance were used to assess auditory-alone speech perception skills. A measure of speech intelligibility was also obtained to assess the speech production abilities of these children. RESULTS: A measure of audiovisual gain, "Ra," was computed using sentence recognition scores in auditory-alone and audiovisual conditions. Another measure of audiovisual gain, "Rv," was computed using scores in visual-alone and audiovisual conditions. The results indicated that children who were better at recognizing isolated spoken words through listening alone were also better at combining the complementary sensory information about speech articulation available under audiovisual stimulation. In addition, we found that children who received more benefit from audiovisual presentation also produced more intelligible speech, suggesting a close link between speech perception and production and a common underlying linguistic basis for audiovisual enhancement effects. Finally, an examination of the distribution of children enrolled in Oral Communication (OC) and Total Communication (TC) indicated that OC children tended to score higher on measures of audiovisual gain, spoken word recognition, and speech intelligibility. CONCLUSIONS: The relationships observed between auditory-alone speech perception, audiovisual benefit, and speech intelligibility indicate that these abilities are not based on independent language skills, but instead reflect a common source of linguistic knowledge, used in both perception and production, that is based on the dynamic, articulatory motions of the vocal tract. The effects of communication mode demonstrate the important contribution of early sensory experience to perceptual development, specifically, language acquisition and the use of phonological processing skills. Intervention and treatment programs that aim to increase receptive and productive spoken language skills, therefore, may wish to emphasize the inherent cross-correlations that exist between auditory and visual sources of information in speech perception. PMID- 11409861 TI - Scandinavian CDG-Ia patients: genotype/phenotype correlation and geographic origin of founder mutations. AB - Congenital disorders of glycosylation type Ia, (previous name carbohydrate deficient glycoprotein syndrome type Ia; CDG-Ia) is an inherited disorder of the glycosylation of certain glycoproteins. The defect is caused by mutations in the phosphomannomutase 2 (PMM2) gene located in chromosome region 16p13. The purpose of this study was twofold: (1) to investigate the possible correlation between certain genotypes and the phenotype of the patients and their PMM activity, and (2) to study further the founder origin of the Scandinavian mutations. Sixty-four CDG-Ia patients were studied. Regardless of mutation combination, the patients showed the basic neurological symptoms associated with CDG-Ia. However, patients carrying the mutation 548T-->C had less severe disease, e.g., no pericardial effusions, malnutrition, or clinical coagulation disturbances. Liver dysfunction and peripheral neuropathy were milder. In contrast, patients carrying mutation 691G-->A showed a high incidence of severe malnutrition and hepatopathy, and they had the highest mortality including affected siblings. Heterozygotes for the two most common mutations (422G-->A and 357C-->A) displayed a phenotype of variable severity sometimes leading to early death. PMM activity showed no correlation with either genotype or phenotype but was reduced in most patients. There was a pronounced geographic clustering for some of the Scandinavian mutations. For example, 548T-->C was almost exclusively found in patients stemming from southeastern parts of Sweden, whereas 26G-->A was found to cluster in a region in the most southern parts of Sweden, suggesting that these mutations originated in these two regions separately as founder mutations. The most frequent mutation (422G-->A) did not show a specific geographic focus. The widespread 422G-->A mutation is probably an older mutation, although haplotype data from intragenic polymorphisms indicate that this mutation also arose only once. The detailed information of the origin of mutations and their respective associated phenotypic pattern should enable improvements to be made regarding tools for genetic counseling and for prenatal diagnoses in CDG-Ia families. PMID- 11409862 TI - Delineation of the critical interval for the familial exudative vitreoretinopathy gene by linkage and haplotype analysis. AB - Familial exudative vitreoretinopathy (FEVR) is an ocular disorder characterized by deficient vascularization of the peripheral retina and causes visual loss attributable to various types of retinal detachment. The locus of the gene responsible for the autosomal dominant form of FEVR (EVR1) has been assigned to 11q13-23. However, a detailed evaluation of the critical region has not been made. We present the results of linkage analysis of the EVR1 locus on 11q13-23 in 43 individuals belonging to seven unrelated families of Japanese origin. Multipoint analysis has shown that six families out of the seven are linked with 11q13-23 markers. Haplotype analysis reveals that the putative region is probably flanked by polymorphic markers D11S1362 and CHLC.GATA30G01, which are approximately 200 kb apart, although the recombination events in small families such as presented in this study should be interpreted cautiously. PMID- 11409863 TI - DHPLC-based germline mutation screening in the analysis of the VHL tumor suppressor gene: usefulness and limitations. AB - In order to evaluate the sensitivity and specificity of the recently introduced high-throughput method DHPLC (denaturing high performance liquid chromatography) for mutation screening in the VHL tumor suppressor gene, we subjected DNA from 43 unrelated VHL patients with previously sequenced VHL germline mutations to this method. In addition, 36 genomic DNAs of unrelated individuals suspected of being VHL carriers but with unknown germline status were analyzed by DHPLC and sequencing. The aims of the present study were to compare mutation results obtained by direct sequencing and DHPLC, and a comparison of two different DHPLC systems. The sensitivity of DHPLC was tested with two commercial devices and protocols, i.e., the Varian-Helix system and the Wave Nucleic Acid Fragment Analysis system. Both resolved all but one mutation in exons 2 and 3 of the VHL gene. In contrast, the GC-rich exon 1 showed discrepancies in the rate of mutation detection. Whereas the Varian-Helix system detected 10/15 (67%) of the known mutations, the Wave Nucleic Acid Fragment Analysis system detected 13/14 (93%). All three mutations in samples with unknown mutation status were revealed by both systems raising the mutation detection rate to 72% and 94%, respectively. Cases with different substitutions at the same nucleotide showed different elution profiles, but similar elution profiles could be obtained from different mutations. The Wave Nucleic Acid Fragment Analysis system detected most VHL mutations; however, when a 100% detection rate is needed, sequencing is still required and must therefore be the standard VHL mutation detection procedure. Once a family-specific mutation has been established, DHPLC may be suitable for the rapid and cost-effective determination of VHL carrier status in family members. PMID- 11409864 TI - Connexin 26 (GJB2) mutations in the Turkish population: implications for the origin and high frequency of the 35delG mutation in Caucasians. AB - Mutations in the Connexin 26 (GJB2/Cx26) gene are responsible for more than half of all cases of prelingual non-syndromic recessive deafness in many Caucasian populations. To determine the importance of Cx26 mutations as a cause of deafness in Turks we screened 11 families with prelingual non-syndromic deafness, seven (64%) of which were found to carry the 35delG mutation. We subsequently screened 674 Turkish subjects with no known hearing loss and found twelve 35delG heterozygotes (1.78%; 95% confidence interval: 0.9%-3%) but no examples of the 167delT mutation. To search for possible founder effects, we typed chromosomes carrying the 35delG mutation for closely linked polymorphic markers in samples from Turkey and United States and compared the allele frequencies with those of hearing subjects. The data showed a modest degree of disequilibrium in both populations. Analyses of two pedigrees from Turkey demonstrated both conserved and different haplotypes, suggesting possible founder effects and multiple origins of the 35delG mutation. PMID- 11409865 TI - Mutation analysis of 4 candidate genes for hereditary neuralgic amyotrophy (HNA). AB - Hereditary neuralgic amyotrophy (HNA) is a rare autosomal dominant disorder. It is characterised by recurrent episodes of focal neuropathy involving the brachial plexus. Genetic linkage analysis has mapped HNA to chromosome 17q25 within a 3.5 cM interval flanked by the short tandem repeat markers D17S785 and D17S802. Here, we report the mutation analysis of four candidate genes. Mutation analysis was performed on the complete coding regions of these genes. Several exonic and intronic single nucleotide polymorphisms were detected. However, no disease causing mutations were found, indicating that these genes are most probably not involved in the pathogenesis of HNA. In addition, we have characterised and localised a putative pseudogene of the SEC14-like 1 gene. PMID- 11409866 TI - Refinement of the chromosome 5p locus for craniometaphyseal dysplasia. AB - Craniometaphyseal dysplasia--Jackson type (CMDJ) is an autosomal dominant bone dysplasia with hyperostosis and sclerosis of the skull and abnormal modelling of the metaphyses. In a large German pedigree, a locus for CMDJ has been mapped previously to the short arm of chromosome 5 (5p15.2-p14.1), defining a 19-cM disease interval between markers D5S2004 and D5S502. Analysis of a large Australian pedigree together with a second German family confirms linkage to the same region. Obligate recombinations in the new families and confirmation of a supposed recombination in the previously reported German kindred have enabled us to narrow the critical region down to approximately 4 cM between markers D5S1987 and D5S1991. PMID- 11409867 TI - Oto-facio-cervical (OFC) syndrome is a contiguous gene deletion syndrome involving EYA1: molecular analysis confirms allelism with BOR syndrome and further narrows the Duane syndrome critical region to 1 cM. AB - Branchio-oto-renal (BOR) syndrome is an autosomal dominant disorder involving hearing loss, branchial defects, ear pits and renal abnormalities. Oto-facio cervical (OFC) syndrome is clinically similar to BOR syndrome, with clinical features in addition to those of BOR syndrome. Mutations in the EYA1 gene (localised to 8q13.3) account for nearly 70% of BOR syndrome cases exhibiting at least three of the major features. Small intragenic deletions of the 3' region of the gene have also been reported in patients with BOR syndrome. We have developed a fluorescent quantitative multiplex polymerase chain reaction for three 3' exons (7, 9 and 13) of the EYA1 gene. This dosage assay, combined with microsatellite marker analysis, has identified de novo deletions of the EYA1 gene and surrounding region in two patients with complex phenotypes involving features of BOR syndrome. One patient with OFC syndrome carried a large deletion of the EYA1 gene region, confirming that OFC syndrome is allelic with BOR syndrome. Microsatellite analysis has shown that comparison of the boundaries of this large deletion with other reported rearrangements of the region reduces the critical region for Duane syndrome (an eye movement disorder) to between markers D8S553 and D8S1797, a genetic distance of approximately 1 cM. PMID- 11409868 TI - Molecular and functional characterisation of mild MCAD deficiency. AB - We report a novel mild variant of medium-chain acyl-CoA dehydrogenase deficiency (MCADD) diagnosed in four infants who, in neonatal screening, showed abnormal acylcarnitine profiles indicative of MCADD. Three patients showed completely normal urinary organic acids and phenylpropionic acid loading tests were normal in all four patients. Enzyme studies showed residual MCAD activities between "classical" MCADD and heterozygotes. ACADM gene analysis revealed compound heterozygosity for the common mutation K329E and a novel mutation, Y67H, in two cases, and homozygosity for mutation G267R and the novel mutation S245L, respectively, in two children of consanguineous parents. As in other metabolic disorders, the distinction between "normal" and "disease" in MCAD deficiency is blurring into a spectrum of enzyme deficiency states caused by different mutations in the ACADM gene potentially influenced by factors affecting intracellular protein processing. PMID- 11409869 TI - Cloning and characterization of the human GFRA2 locus and investigation of the gene in Hirschsprung disease. AB - The glial-cell-line-derived neurotrophic factor (GDNF) family receptors alpha (GFRalpha) are cell surface bound glycoproteins that mediate interactions of the GDNF ligand family with the RET receptor. These interactions are crucial to the development of the kidney and some peripheral nerve lineages. In humans, mutations of RET or RET ligands are associated with the congenital abnormality Hirschsprung disease (HSCR) in which nerves and ganglia of the hind gut are absent. As the GFRalpha family are required for normal activation of the RET receptor, they are also candidates for a role in HSCR. The GFRA2 gene, which is required for the development of the myenteric nerve plexus, is an excellent candidate gene for HSCR. In this study, we cloned the human GFRA2 locus, characterized the gene structure, and compared it with other GFRA family members. We further investigated the GFRA2 gene for mutations in a panel of HSCR patients. GFRA2 has nine coding exons that are similar in size and organization to those of other GFRA family genes. We identified six sequence variants of GFRA2, four of which did not affect the amino acid sequence of the GFRalpha-2 protein. Two further changes that resulted in amino acid substitutions were found in exon 9 and were predicted to lie in the amino acid sequence encoding the glycosylphosphatidylinositol-linkage signal of GFRalpha-2. There was no difference in frequency of any of the sequence variants between control and HSCR populations. Our data indicate that members of the GFRA gene family are closely related in intron/exon structure and in sequence. We have not detected any correlation between sequence variants of GFRA2 and the HSCR phenotype. PMID- 11409870 TI - Somatic NF1 mutational spectrum in benign neurofibromas: mRNA splice defects are common among point mutations. AB - Neurofibromas, benign tumors that originate from the peripheral nerve sheath, are a hallmark of neurofibromatosis type 1 (NF1). Although loss of heterozygosity (LOH) is a common phenomenon in this neoplasia, it only accounts for part of the somatic NF1 mutations found. Somatic point mutations or the presence of "two hits" in the NF1 gene have only been reported for a few neurofibromas. The large size of the NF1 gene together with the multicellular composition of these tumors has greatly hampered their molecular characterization. Here, we present the somatic NF1 mutational analysis of the whole set of neurofibromas studied by our group and consisting in 126 tumors derived from 32 NF1 patients. We report the identification of 45 independent somatic NF1 mutations, 20 of which are reported for the first time. Different types of point mutations together with LOH affecting the NF1 gene and its surrounding region or extending along the 17q arm have been found. Among point mutations, those affecting the correct splicing of the NF1 gene are common, coinciding with results reported on germline NF1 mutations. In most cases, we have been able to confirm that both copies of the NF1 gene are inactivated. We have also found that both somatic and germline mutations can be expressed at the RNA level in the neoplastic cells. Furthermore, we have observed that the study of more than one tumor derived from the same patient is useful for the identification of the germline mutation. Finally, we have noticed that the culture of neurofibromas and their fibroblast clearance facilitates LOH detection in cases in which it is difficult to determine. PMID- 11409871 TI - Evidence for a prostate cancer linkage to chromosome 20 in 159 hereditary prostate cancer families. AB - Prostate cancer is the most common malignancy diagnosed in men in the US. Genetic susceptibility to prostate cancer has been well documented. A region at chromosome 20q13 (HPC20) has been reported to be linked to a prostate cancer susceptibility gene. To confirm this finding, we genotyped 16 markers spanning approximately 95 cM on chromosome 20 in 159 hereditary prostate cancer (HPC) families. Positive (but not statistically significant) linkage scores were observed from 20pter to 20q11, with the highest non-parametric linkage (NPL) score for the complete dataset of 1.02 (P=0.15) being observed at D20S195 at 20q11. Evidence for linkage from parametric analyses with a dominant or a recessive model was weak. Interestingly, consistent with the original findings of linkage to 20 g higher linkage scores were observed in the subsets of families with a later age at diagnosis (> or =65 years; n=80, NPL=1.94, P=0.029 at D20S186), fewer than five affected family members (n=69, NPL=1.74, P=0.037 at D20S889), or without male-to-male disease transmission (n=60, NPL=1.01, P=0.15 at D20S117). The region with positive linkage scores spanned approximately 60 cM from 20pter to 20q11 in these subsets of families. Our results are consistent with a prostate cancer susceptibility locus on chromosome 20. PMID- 11409872 TI - Topoisomerase-I- and Alu-mediated genomic deletions of the APC gene in familial adenomatous polyposis. AB - Germline mutation in the adenomatous polyposis coli (APC) gene results in familial adenomatous polyposis (FAP), a heritable form of colorectal cancer. We have previously reported two novel mutations that delete exons 11 and 14 of the APC gene, respectively, at the cDNA level without any splice junction defects at the genomic level. We describe here the precise breakpoints of the two mutations and the possible mechanisms leading to the genomic rearrangement. The first rearrangement is most likely a topoisomerase-I-mediated non-homologous recombination resulting in a 2-kb deletion that deletes exon 11 of the APC gene. Both 5' and 3' breakpoints have two topoisomerase I recognition sites and runs of pyrimidines within the 10-bp sequences in their vicinity. Further, the 3' breakpoint has an adenine-thymidine-rich region. This is probably the first report of a topoisomerase-I-mediated germline mutation in a tumor suppressor gene. The second rearrangement is most likely an Alu-Alu homologous recombination resulting in a 6-kb deletion encompassing exon 14. The Alu elements at the 5' and 3' breakpoints include the 26-bp core sequence thought to stimulate recombination. In both rearrangements, partial sequences from the long interspersed nuclear element family are in the vicinity of the breakpoints. Other than serving as markers for regions of DNA damage, their precise role in the recombination events, if any, is unclear. Both deletions result in truncated APC proteins missing the beta-catenin- and axin-binding domains, resulting in severe polyposis and cancer. PMID- 11409873 TI - Transport of fluorescein in MDCKII-MRP1 transfected cells and mrp1-knockout mice. AB - The multidrug resistant-associated protein 1 (MRP1) is a membrane-bound transport protein that is involved in the efflux of organic anions and has been implicated in multidrug resistance in cancer. MRP1 has also been reported to be ubiquitously expressed in normal tissues, including the brain. The presence of functional organic anion transporters in the blood-brain and blood-CSF barriers that influence the distribution of various compounds to the brain has long been known. The purpose of this study was to examine the role of MRP1 in the brain distribution of a model organic anion, fluorescein. The substrate specificity of MRP1 for fluorescein was initially determined by examining the accumulation of fluorescein in MDCKII MRP1-transfected cells. The distribution of fluorescein in the brain was then examined in wild-type and mrp1 gene knockout mice. The results show that in MDCKII MRP1-transfected cells, the accumulation of fluorescein was significantly lower (about 40% lower) than that in wild-type MDCKII cells. MRP1 inhibitors such as probenecid, MK-571, and LY402913 enhanced fluorescein accumulation in MDCKII MRP1-transfected cells to a greater extent than in wild type MDCKII cells. In an in vivo study, after intravenous injection of fluorescein, the fluorescein brain-to-plasma concentration ratio in mrp1 knockout mice was not significantly different than that in wild-type mice. However, when probenecid was co-administered with fluorescein in wild-type mice, the fluorescein brain-to-plasma ratio was significantly increased (1.5-fold). These findings suggest that fluorescein is a substrate for MRP1. Furthermore, the in vivo study also suggests that MRP1 has a limited role in the transport and distribution of fluorescein in the brain. Therefore, other organic anion transport proteins, including the various isoforms of the MRP family, may be responsible for the accumulation and transport of organic anions in the brain. PMID- 11409874 TI - Factors contributing to the fusogenic potency of foamy virus. AB - Three model peptides have been studied in an effort to understand the molecular basis for the fusogenic potency of foamy virus. These peptides corresponded to a 23 amino acid helical segment close to the amino terminus, a shortened 17 amino acid, more hydrophobic homolog of this peptide, and an 18-amino-acid peptide close to or within the transmembrane domain. The peptides have a conformation containing both alpha-helical and beta-structure in aqueous solution but are predominantly alpha-helical in solutions of trifluoroethanol, as assessed by circular dichroism. In common with other viruses, the most fusogenic peptide was the one closest to the amino terminus. However, unlike several other fusion peptides that have been studied previously, this peptide did not promote increase negative membrane curvature as assessed by effects of the peptide on lipid polymorphism. Nevertheless, the foamy virus fusion peptide promotes membrane fusion, apparently by a mechanism that is independent of changes in membrane curvature. We demonstrate that there is a synergistic action in the promotion of membrane fusion between the peptide from the amino terminal region and the one from the region adjacent to the transmembrane segment. PMID- 11409875 TI - Dys-regulation of effector CD4+ T cell function by the V3 domain of the HIV-1 gp120 during antigen presentation. AB - It was recently demonstrated that the semiconserved domain of the V3 region of the HIV-1 surface glycoprotein gp120 can induce an activation-apoptosis phenomenon to memory CD4+ cells from healthy individuals. Studying the effects of V3 on the interaction of antigen presentation between monocyte-derived macrophages and resting memory CD4+ T cells, we observed that V3 affects both cell populations. Macrophages exposed to composite liposomes containing V3 on the surface and tetanus toxoid (TT) as the recall antigen entrapped in the aqueous phase (lipoV3/TT liposomes) were able to activate CD4+ T cells during primary stimulation, but not after restimulation nine days later. Unstimulated macrophages or macrophages exposed to soluble TT responded to second stimuli, lipoV3/TT liposomes, and soluble TT in activating CD4+ T cells. Soluble TT activated CD4+ T cells could be restimulated by soluble TT but not by lipoV3/TT liposomes, whereas lipoV3/TT liposome-activated CD4+ T cells became unresponsive to a second stimulus. These results show that resting memory CD4+ cells activated by macrophages presenting the recall antigen together with V3 become unresponsive to restimulation. PMID- 11409876 TI - Nuclear extracellular signal-regulated kinase 2 phosphorylates p53 at Thr55 in response to doxorubicin. AB - In this study, we showed that nuclear ERK2 phosphorylates p53 at Thr55 in response to doxorubicin. p53 was found to physically interact with ERK2 as evidenced by Western blotting of ERK2 coimmunoprecipitated complex. The gene fragment encoded for N-terminal 68 amino acids was subcloned and fused with 6 His. Each serine or threonine site in this fragment, the possible phosphorylation site, was mutated to alanine. The recombinant proteins were used as substrates in ERK2 kinase assay. The results show that ERK2 phosphorylated p53 at Thr55. Further, electromobility shift assay showed that the phosphorylation of p53 by nuclear ERK2 was closely related to the transactivating activity of p53. These findings suggest that ERK2 may play a role in response to DNA damage via interaction with p53. PMID- 11409877 TI - Cloning and molecular characterization of two splice variants of a new putative member of the Siglec-3-like subgroup of Siglecs. AB - The sialic acid binding immunoglobulin-like lectin (Siglec) family is a recently described member of the immunoglobulin superfamily. Within this Siglec family there exists a subgroup of molecules which bear a very high degree of homology with the molecule Siglec-3 (CD33), and has thus been designated the Siglec-3-like subgroup of Siglecs. The members of this subgroup have been localized to chromosome 19q13.4, through both in situ hybridization and precise genomic mapping at the nucleotide level. Through the positional cloning approach we have identified and characterized a Siglec-like gene (SLG), a putative novel member of the Siglec-3-like subgroup of Siglecs. We have characterized the complete genomic structure of SLG, as well as two alternative splice variants, and determined its chromosomal localization. The short isoform, SLG-S, consists of seven exons, with six intervening introns, while the longer isoform, SLG-L, consists of eight exons and seven intervening introns. The SLG gene is localized 32.9 kb downstream of Siglec-8 on chromosome 19q13.4. The putative SLG-S and SLG-L proteins, of 477 and 595 amino acid residues, respectively, show extensive homology to many members of the Siglec-3-like subgroup. This high degree of homology is conserved in the extracellular Ig-like domains, as well as in the cytoplasmic tyrosine-based motifs. Interestingly, the SLG-L protein contains two N-terminal V-set Ig-like domains, as opposed to SLG-S and other Siglec-3-like subgroup members which contain only one such domain. Through RT-PCR we have examined the expression profile of both SLG splice variants in a panel of human tissues and have found that SLG-S is highly expressed in spleen, small intestine and adrenal gland, while SLG-L exhibits high levels of expression in spleen, small intestine, and bone marrow. This gene is quite likely the latest novel member of the CD33-like subgroup of Siglecs, and given its high degree of homology, it may also serve a regulatory role in the proliferation and survival of a particular hematopoietic stem cell lineage, as has been found for CD33 and Siglec7. PMID- 11409878 TI - Molecular characterization, tissue expression, and mapping of a novel Siglec-like gene (SLG2) with three splice variants. AB - The sialic acid binding immunglobulin-like lectin (Siglec) family is a recently described member of the immunoglobulin superfamily. Within the Siglec family, there exists a subgroup, which bears a high degree of homology with the molecule CD33 (Siglec-3), and has thus been designated the CD33-like subgroup of Siglecs. Members of this subgroup have been localized to chromosome 19q13.4. Through the positional candidate approach, we identified a novel potential member of this subgroup of Siglecs. We have characterized the complete genomic structure of this gene, determined its chromosomal localization, its homology to other members of the Siglec family, and its tissue expression profile. This new Siglec-like gene is comprised of 11 exons, with 10 intervening introns, and is localized 278 kb telomeric to Siglec-9 and 35 kb centromeric to Siglec-8 and on chromosome 19q13.4. The coding region consists of 2094 base pairs, and encodes for a putative 76.6 kDa protein. All Siglec-conserved structural features, including V set domain, three C-set domains, transmembrane domain, ITIM and SLAM motifs, were found in this Siglec-like gene. Also, it has the conserved amino acids essential for sialic acid binding. The Siglec-like gene has 40-66% homology with members of the CD33-like subgroup, including Siglecs 5-9. Through RT-PCR we have examined the expression profile of this new gene in a panel of human tissues and found it to be primarily expressed in the bone marrow, spleen, brain, small intestine, colon, and spinal cord. We were also able to identify three different splice variants of the new gene. This gene may represent the latest novel member of the CD33-like subgroup of Siglecs, and, given its high degree of homology, it may also serve a regulatory role in the proliferation and survival of a particular hematopoietic stem cell lineage, as has been found for CD33 and Siglec-7. PMID- 11409879 TI - Molecular cloning and sequencing of a novel cDNA encoding for a protein involved in human sperm function. AB - A cDNA encoding for an antigen, designated as NZ-3, was cloned and sequenced from human testis. The 1481-bp NZ-3 cDNA yielded an open reading frame (ORF) of 231 amino acids (aa) with the first ATG, Met start codon at nucleotide (nt) 104 and the stop codon TGA at nt 797. Extensive computer search indicated it to be a novel cDNA/protein. The ORF of NZ-3 cDNA was subcloned into pGEX-1lambdaT vector and expressed in glutathione S-transferase gene fusion system. The expressed recombinant protein had a molecular size of approximately 25 kDa, and the rabbit antibodies (Ab) against the recombinant antigen recognized a specific protein band of 63 +/- 3 kDa in the human testis extract. The NZ-3 antigen was located on the acrosomal and tail regions of human sperm cell and the NZ-3 Ab significantly (P < 0.001) inhibited human sperm capacitation and/or acrosome reaction. The novel recombinant NZ-3 antigen may find applications in immunocontraception and in specific diagnosis of human infertility. PMID- 11409880 TI - The imidazoline RX871024 stimulates insulin secretion in pancreatic beta-cells from mice deficient in K(ATP) channel function. AB - Effects of the imidazoline compound RX871024 on cytosolic free Ca(2+) concentration ([Ca(2+)]i) and insulin secretion in pancreatic beta-cells from SUR1 deficient mice have been studied. In beta-cells from wild-type mice RX871024 increased [Ca(2+)]i by blocking ATP-dependent K(+)-current (K(ATP)) and inducing membrane depolarization. In beta-cells lacking a component of the K(ATP)-channel, SUR1 subunit, RX871024 failed to increase [Ca(2+)]i. However, insulin secretion in these cells was strongly stimulated by the imidazoline. Thus, a major component of the insulinotropic activity of RX871024 is stimulation of insulin exocytosis independently from changes in K(ATP)-current and [Ca(2+)]i. This means that effects of RX871024 on insulin exocytosis are partly mediated by interaction with proteins distinct from those composing the K(ATP)-channel. PMID- 11409881 TI - TASK-5, a new member of the tandem-pore K(+) channel family. AB - TASKs are members of the recently identified K(+) channel family (KCNKx). Four TASKs (TASK1-4) identified so far form functional K(+) channels and encode background K(+) channels in various cell types. Recently, another member (TASK-5) was identified in the human genome. We cloned it and studied its tissue expression and functional properties. TASK-5 shares 51% amino acid identity with TASK-1 and TASK-3. Northern blot analysis showed that TASK-4 mRNA was expressed primarily in the adrenal gland and pancreas. Single nucleotide polymorphism (SNP) was found at amino acid position 95 that normally forms part of the K(+) channel selectivity filter. Neither form of TASK-5 showed channel activity when transfected in COS-7 cells. Exchange of C-termini of TASK-3 and TASK-5 failed to generate whole-cell currents. Thus, TASK-5 is a new member of the tandem-pore K(+) channel family but does not produce a functional plasma membrane K(+) current by itself. PMID- 11409882 TI - A novel recombinant basic fibroblast growth factor and its secretion. AB - Basic fibroblast growth factor (FGF-2) is a pleiotropic mitogen which plays an important role in cell growth, differentiation, migration, and survival in different cells and organ systems. Recently, several clinical applications for FGF-2 gene transfer are being evaluated in wound healing and collateral artery development to relieve myocardial and peripheral ischemia due to the ability of FGF-2 to regulate the growth and function of vascular cells. However, FGF-2 lacks a classical hydrophobic secretion signal peptide, the FGF-2 chimeras containing various signal sequences have been explored. In this study, a novel recombinant 4sFGF-2 was constructed by replacing nine residues from the amino-terminus of native FGF-2 (Met1 to Leu9) with eight amino acid residues of signal peptide of FGF-4 (Met1 to Ala8) to better increase the secretion level of FGF-2. When the recombinant FGF-2 gene, cloned into the expression vector with CMV promoter, was expressed in COS-7 cells, the recombinant 4sFGF-2 was highly secreted into the media. The secreted 4sFGF-2 showed the same biological activity as the native FGF 2 in the dose-response effects on DNA synthesis and cell growth of rat aortic smooth muscle cells (RASMCs) and NIH3T3 cells. The 4sFGF-2 also was able to activate MAPK as wild FGF-2 in RASMCs. These results indicate that a novel recombinant 4sFGF-2 may be useful as clinical applicability of angiogenic growth factor gene transfer. PMID- 11409883 TI - An enzymatically active chimeric protein containing the hydrophilic form of NADPH cytochrome P450 reductase fused to the membrane-binding domain of cytochrome b5. AB - The microsomal flavoprotein NADPH-cytochrome P450 reductase (CPR) contains an N terminal hydrophobic membrane-binding domain required for reconstitution of hydroxylation activities with cytochrome P450s. In contrast, cytochrome b5 (b5) contains a C-terminal hydrophobic membrane-binding domain required for interaction with P450s. We have constructed, expressed and purified a chimeric flavoprotein (hdb5-CPR) where the C-terminal 45 amino acid residues of b5 have replaced the N-terminal 56 amino acid domain of CPR. This hybrid flavoprotein retains the catalytic properties of the native CPR and is able to reconstitute fatty acid and steroid hydroxylation activities with CYP4A1 and CYP17A. However hdb5-CPR is much less effective than CPR for reconstituting activity with CYP3A4. We conclude that differences on the surface of the P450s reflect unique and specific information essential for the recognition needed to establish reactions of intermolecular electron transfer from the flavoprotein CPR. PMID- 11409884 TI - Preheat treatment for Mycobacterium tuberculosis Hsp16.3: correlation between a structural phase change at 60 degrees C and a dramatic increase in chaperone-like activity. AB - The in vitro chaperone-like activity of Mycobacterium tuberculosis small heat shock protein Hsp16.3 was found to be dramatically enhanced to the same extent after preheat treatment at or over 60 degrees C. Structural analysis using gel filtration, native pore-gradient PAGE, nondenaturing PAGE, and far-UV CD spectroscopy consistently revealed no significant difference between the native and the preheated Hsp16.3 proteins. However, near-UV CD spectroscopy clearly demonstrated that the tertiary structure of preheated Hsp16.3 is quite similar to its native conformation, with a minor but significant difference. Further analysis using differential scanning calorimetry indicated that Hsp16.3 exhibited a structural transition near 60 degrees C. All these results together indicate that Hsp16.3 suffers a phase change at approximately 60 degrees C, which seem to remove a structural energy barrier for the protein to refold to a conformational status with increased chaperone-like activity. PMID- 11409885 TI - Conserved base-pairings between C266-A268 and U307-G309 in the P7 of the Tetrahymena ribozyme is nonessential for the in vitro self-splicing reaction. AB - P7 is highly conserved in Group I self-splicing intron ribozymes. This region is known to coordinate metal ions and bind a cofactor guanosine required for the self-splicing. To further investigate the fundamental role of the corresponding region in the Tetrahymena ribozyme, we attempted to identify minimal requirements for the base-paired region excluding the guanosine binding site. We discovered that a variety of sequences are eligible and its derivatives possessing extra nucleotide(s) can still conduct the first step of splicing, indicating that no particular base-pairing is essential in this region for conducting the reaction in vitro. The results provide two hypotheses for the fundamental role of this region: (i) if the region contains element(s) that are strictly required in the catalysis, they are not necessarily tightly fixed in the ribozyme and (ii) if not, its fundamental role may simply be to coordinate neighboring regions that are directly involved in the catalysis. PMID- 11409886 TI - High-level production of functional muscle alpha-tropomyosin in Pichia pastoris. AB - Although numerous studies have reported the production of skeletal muscle alpha tropomyosin in E. coli, the protein needs to be modified at the amino terminus in order to be active. Without these modifications the protein does not bind to actin, does not exhibit head-to-tail polymerization, and does not inhibit the actomyosin Mg(2+)-ATPase in the absence of troponin. On the other hand, the protein produced in insect cells using baculovirus as an expression vector (Urbancikova, M., and Hitchcock-DeGregori, S. E., J. Biol. Chem., 269, 24310 24315, 1994) is only partially acetylated at its amino terminal and therefore is not totally functional. In an attempt to produce an unmodified functional recombinant muscle alpha-tropomyosin for structure-function correlation studies we have expressed the chicken skeletal alpha-tropomyosin cDNA in the yeast Pichia pastoris. Recombinant protein was produced at a high level (20 mg/L) and was similar to the wild type muscle protein in its ability to polymerize, to bind to actin and to regulate the actomyosin S1 Mg(2+)-ATPase. PMID- 11409887 TI - Induction of the oxLDL receptor LOX-1 by endothelin-1 in human endothelial cells. AB - In this study, we analyzed the effect of endothelin-1 (ET-1) on expression of the lectin-like oxidized low-density lipoprotein (oxLDL) receptor-1 LOX-1 and on oxLDL uptake in primary cultures of human umbilical vein endothelial cells (HUVEC). LOX-1 mRNA was quantified by standard-calibrated competitive RT-PCR, LOX 1 protein expression by Western analysis and endothelial oxLDL uptake using DiI labeled oxLDL. ET-1 induces LOX-1 mRNA expression, reaching its maximum after 1 h (160 +/- 14% of control, 100 nM ET-1, P < 0.05). This increased ET-1-mediated LOX 1 mRNA expression could be inhibited by endothelin receptor B antagonist BQ-788. In addition, ET-1 stimulates LOX-1 protein expression and oxLDL uptake in HUVEC. The augmented oxLDL uptake by ET-1 is mediated by endothelin receptor B, but not by protein kinases. These data support a new pathophysiological mechanism how locally and systemically increased ET-1 levels could promote LOX-1-mediated oxLDL uptake in human endothelial cells and the development and progression of endothelial dysfunction and atherosclerosis. PMID- 11409888 TI - Hydrogen peroxide is a novel inducer of connective tissue growth factor. AB - Connective tissue growth factor (CTGF) has recently been described as a fibrogenic factor and is greatly induced by various extracellular stimuli, such as transforming growth factor-beta (TGF-beta), dexamethasone, and serotonin. CTGF induces collagen type I and fibronectin, and the deposition of such molecules leads to fibrotic disease in many tissues. Intracellular reactive oxygen species (ROS) are generated by extracellular stress conditions and are produced as by products of cellular metabolism. Imbalanced cellular redox status is a potent pathogenic factor that leads to various degenerative diseases, including tissue fibrosis. Since CTGF is believed to play a crucial role in fibrotic disease formation in many tissues, we examined the role of ROS in CTGF gene expression in human lens epithelial cell line B3. The results showed that CTGF was induced by reactive oxygen species such as hydrogen peroxide and hydroxyl radicals. Next, we examined whether CTGF induction by ROS is via newly synthesized TGF-beta. The results showed that ROS directly induced CTGF mRNA not via the increased TGF-beta synthesis or activation. Next, we treated AG490, which is the well-known inhibitor of Janus kinase (JAK), with hydrogen peroxide. AG490 abrogated the CTGF induction by ROS in a dose-dependent manner. The results suggest that JAK-2/-3 seems to be involved in the enhanced CTGF mRNA expression by hydrogen peroxide. In this report, we present that hydrogen peroxide is a novel inducer of CTGF gene expression and that JAK-2/-3 activation seems to play a role in CTGF induction. PMID- 11409889 TI - Enhanced post-ischemic liver injury in iNOS-deficient mice: a cautionary note. AB - The objective of this study was to assess the role of inducible nitric oxide synthase (iNOS) in ischemia- and reperfusion (I/R)-induced liver injury. We found that partial hepatic ischemia involving 70% of the liver resulted in a time dependent increase in serum alanine aminotransferase (ALT) levels at 1-6 h following reperfusion. Liver injury at 1, 3, and 6 h post-ischemia was not due to the infiltration of neutrophils as assessed by tissue myeloperoxidase (MPO) activity and histopathology. iNOS-deficient mice subjected to the same duration of ischemia and reperfusion showed dramatic and significant increases in liver injury at 3 but not 6 h following reperfusion compared to their wild type controls. Paradoxically, iNOS mRNA expression was not detected in the livers of wild type mice at any point during the reperfusion period and pharmacological inhibition of iNOS using L-N(6)(iminoethyl)-lysine (L-NIL) did not exacerbate post-ischemic liver injury at any time post-reperfusion. These data suggest that iNOS deficiency produces unanticipated genetic alterations that renders these mice more sensitive to liver I/R-induced injury. PMID- 11409890 TI - FGF-23 inhibits renal tubular phosphate transport and is a PHEX substrate. AB - Oncogenic osteomalacia (OOM), X-linked hypophosphatemia (XLH), and autosomal dominant hypophosphatemic rickets (ADHR) are phenotypically similar disorders characterized by hypophosphatemia, decreased renal phosphate reabsorption, normal or low serum calcitriol concentrations, normal serum concentrations of calcium and parathyroid hormone, and defective skeletal mineralization. XLH results from mutations in the PHEX gene, encoding a membrane-bound endopeptidase, whereas ADHR is associated with mutations of the gene encoding FGF-23. Recent evidence that FGF-23 is expressed in mesenchymal tumors associated with OOM suggests that FGF 23 is responsible for the phosphaturic activity previously termed "phosphatonin." Here we show that both wild-type FGF-23 and the ADHR mutant, FGF-23(R179Q), inhibit phosphate uptake in renal epithelial cells. We further show that the endopeptidase, PHEX, degrades native FGF-23 but not the mutant form. Our results suggest that FGF-23 is involved in the pathogenesis of these three hypophosphatemic disorders and directly link PHEX and FGF-23 within the same biochemical pathway. PMID- 11409891 TI - APC senses cell-cell contacts and moves to the nucleus upon their disruption. AB - The adenomatous polyposis coli (APC) tumor suppressor protein is involved in the Wnt/wingless pathway, modulating beta-catenin activity. We report the development of a highly specific, chemically synthesized oligobody (oligonucleotide-based synthetic antibody), directed against the N-terminal region of APC. Using this reagent, we found that within 16 h of disrupting HT-29 cell-cell contacts by harvesting cells with trypsin/EDTA treatment and replating, APC was translocated from the cytoplasm to the nucleus. Five days after plating the cells, when the cells had returned to their normal confluent phenotype and cell-cell contacts were reestablished, APC returned to the cytoplasm. These results suggest that APC functions as part of a "sensor" system, and responds to the loss of cell-cell contacts by moving to the nucleus, and returning to the cytoplasm when the contacts are fully restored. PMID- 11409892 TI - The sea urchin sns insulator blocks CMV enhancer following integration in human cells. AB - Insulators are a new class of genetic elements that attenuate enhancer function directionally. Previously, we characterized in sea urchin a 265-bp-long insulator, termed sns. To test insulator activity following stable integration in human cells, we placed sns between the CMV enhancer and a tk promoter upstream of a GFP transgene of plasmid or retroviral vectors. In contrast to controls, cells transfected or transduced with insulated constructs displayed a barely detectable fluorescence. Southern blot and PCR ruled out vector rearrangement following integration into host DNA; RNase protection confirmed the enhancer blocking activity. Finally, we demonstrate that two cis-acting sequences, previously characterized in sea urchin, are also specific binding sites for human proteins. We conclude that sns interferes with enhancer promoter interaction also in a human chromatin context. The relatively small size, evolutionary conservation and apparent lack of enhancer specificity might result useful in gene transfer experiments in human cells. PMID- 11409893 TI - Abnormal uterus with polycysts, accumulation of uterine prostaglandins, and reduced fertility in mice heterozygous for acyl-CoA synthetase 4 deficiency. AB - Arachidonate released by various stimuli is rapidly reesterified into membrane phospholipids initiated by acyl-CoA synthetase (ACS) and subsequent acyl-transfer reactions. ACS4 is an arachidonate-preferring enzyme abundant in steroidogenic tissues and postulated to modulate eicosanoid production. Female mice heterozygous for ACS4 deficiency become pregnant less frequently and produce small litters with extremely low transmission of the disrupted alleles. Striking morphological changes, including extremely enlarged uteri and lumina filled with numerous proliferative cysts of various sizes, were detected in ACS4+/- females. Furthermore, marked accumulation of prostaglandins was seen in the uterus of the heterozygous females. These results indicate that ACS4 modulates female fertility and uterine prostaglandin production. PMID- 11409894 TI - A family of Turandot-related genes in the humoral stress response of Drosophila. AB - The Drosophila Turandot A (TotA) gene was recently shown to encode a stress induced humoral factor which gives increased resistance to the lethal effects of high temperature. Here we show that TotA belongs to a family of eight Tot genes distributed at three different sites in the Drosophila genome. All Tot genes are induced under stressful conditions such as bacterial infection, heat shock, paraquat feeding or exposure to ultraviolet light, suggesting that all members of this family play a role in Drosophila stress tolerance. The induction of the Tot genes differs in important respects from the heat shock response, such as the strong but delayed response to bacterial infection seen for several of the genes. PMID- 11409895 TI - The CASK/Lin-2 Drosophila homologue, Camguk, could play a role in epithelial patterning and in neuronal targeting. AB - Drosophila Camguk (Cmg) is a member of the CAMGUK subfamily of the MAGUK family of proteins which are localized at cell junction and other plasma membrane specialized regions, from worms to mammals. The protein structure of Cmg, as the other CAMGUK proteins, is characterized by only one PDZ domain and an additional CaM kinase domain, similar to CaMKII. While the mammalian ortholog CASKs play an important role in synaptic protein targeting and in synaptic plasticity, the Drosophila Cmg role is unknown. To study its potential role, we reported a detailed analysis of mRNA distribution of the Drosophila cmg gene at cellular and developmental level, during embryonic, larval, pupal and adult stages. The transient cmg transcription in midgut and Malpighian tubules may suggest a potential function in cell junction formation and in epithelial tissue patterning. Interestingly, cmg transcription increases substantially during embryonic neuroblast proliferation, becoming predominant in the developing central nervous system (CNS) during embryonic and postembryonic development stages and in the mature brain. In addition, a high transcriptional level was detected in the eye imaginal discs and in the adult retina, demonstrating a specific and continuous expression of cmg in neuroblasts and photoreceptor neurons, from the onset of cytodifferentiation. Our findings suggest that Cmg could play a potential role in transmembrane protein targeting, particularly in synapses. These observations suggest the existence of a common highly conserved mechanism involved in forming and maintaining proper synaptic protein targeting, which are fundamental features of synaptic plasticity, learning and memory. Through its function, the CaM kinase domain-containing Cmg may be involved in signal transduction cascade. Its potential relation to Calmodulin and CaMKII is discussed. PMID- 11409896 TI - The effect of resonance energy homotransfer on the intrinsic tryptophan fluorescence emission of the bothropstoxin-I dimer. AB - Bothopstoxin-I (BthTX-I) is a homodimeric Lys49-PLA2 homologue from the venom of Bothrops jararacussu in which a single Trp77 residue is located at the dimer interface. Intrinsic tryptophan fluorescence emission (ITFE) quenching by iodide and acrylamide has confirmed that a dimer to monomer transition occurs on reducing the pH from 7.0 to 5.0. Both the monomer and the dimer showed an excitation wavelength-dependent increase in the fluorescence emission maximum, however the excitation curve of the dimer was blue-shifted with respect to the monomeric form. No differences in the absorption or circular dichroism spectra between pH 5.0 and 7.0 were observed, suggesting that this curve shift is due neither to altered electronic ground states nor to exciton coupling of the Trp residues. We suggest that fluorescence resonance energy homotransfer between Trp77 residues at the BthTX-I dimer interface results in excitation of an acceptor Trp population which demonstrates a red-shifted fluorescence emission. PMID- 11409897 TI - Molecular mechanisms of ceramide-mediated CD95 clustering. AB - Receptor clustering has been suggested as a crucial mechanism to initiate receptor signaling. Here we show that ceramide in sphingolipid-rich membrane rafts mediates clustering of CD95. Neutralization of surface ceramide or inhibition of its endogenous generation prevented CD95 clustering. Furthermore, application of ceramide at the cell surface triggered clustering of active but not inactive CD95. Apoptosis was inhibited by neutralization of surface ceramide or inhibition of ceramide release in vitro and in vivo. Thus, we conclude that surface ceramide mediates CD95 clustering, which is required for initiation of apoptosis, at least in some cell types. PMID- 11409898 TI - Low doses of EPO activate MAP kinases but not JAK2-STAT5 in rat vascular smooth muscle cells. AB - Previous reports have shown a direct effect of erythropoietin (Epo) on vascular smooth muscle cells (VSMCs). Our aim was to assess expression of the Epo receptor (EpoR) on VSMCs and to study the activation of two major signaling cascades activated by Epo, namely JAK2/STAT5 and MAPK pathways. All experiments were performed in parallel using the Epo-responsive UT7 cell line. From semiquantitative RT-PCR experiments, VSMCs were estimated to express approximately 30-fold less EpoR mRNA than UT7 cells. Epo-induced phosphorylation of proteins involved in the EpoR/JAK2/STAT5 cascade could not be detected in VSMCs, even using pharmacological doses of Epo (250 IU/ml). In contrast, a strong activation of MAP kinase pathway was detected with as low as 10 IU/ml Epo. We suggest that MAPK activation reflects a physiologically relevant effect of Epo on VSMCs that may be correlated to cell proliferation. PMID- 11409899 TI - Genomic organization of the Shc-related phosphotyrosine adapters and characterization of the full-length Sck/ShcB: specific association of p68 Sck/ShcB with pp135. AB - The Shc gene family is an emerging family, containing at least three members designated Shc/ShcA, Sck/Sli/ShcB, N-Shc/Rai/ShcC in mammals. In this study, we determined the genomic organization of the mouse Shc family. Coding regions of ShcA, B, and C each comprised 12 exons, spanned approximately 6, 20, and 65 kb, and located on chromosome 3, 10, and 13, respectively. Based on this genome analysis, we determined the full-length structure of mouse Sck/ShcB as a 68-kD protein. We found that the 68-kD full-length Sck/ShcB was more efficiently phosphorylated upon EGF treatment than the previously-analyzed CH2-deleted form. We also found that Sck specifically interacted with a 135-kD phosphoprotein (pp135) through its SH2 domain following membrane depolarization. The Sck-pp135 interaction was reduced by Src kinase inhibitors. These results suggest that Sck, but not N-Shc nor Shc, transmit signals in conjunction with pp135 following Src activation and/or calcium entry in the cell. PMID- 11409900 TI - Effects of endothelin-1 on K(+) currents from rat ventricular myocytes. AB - It has been suggested that the positive inotropic effect of the vasoactive peptide hormone, endothelin-1 (ET-1), involves inhibition of cardiac K(+) currents. In order to identify the K(+) currents modulated by ET-1, the outward K(+) currents of isolated rat ventricular myocytes were investigated using whole cell patch-clamp recording techniques. Outward currents were elicited by depolarisation to +40 mV for 200 ms from the holding potential of -60 mV. Currents activated rapidly, reaching a peak (I(pk)) of 1310 +/- 115 pA and subsequently inactivating to an outward current level of 1063 +/- 122 pA at the end of the voltage-pulse (I(late)) (n = 11). ET-1 (20 nM) reduced I(pk) by 247.6 +/- 60.7 pA (n = 11, P < 0.01) and reduced I(late) by 323.2 +/- 43.9 pA (P < 0.001). The effects of ET-1 were abolished in the presence of the nonselective ET receptor antagonist, PD 142893 (10 microM, n = 5). Outward currents were considerably reduced and the effects of ET-1 were not observed when K(+) was replaced with Cs(+) in the experimental solutions; this indicates that ET-1 modulated K(+)-selective currents. A double-pulse protocol was used to investigate the inactivation of the currents. The voltage-dependent inactivation of the currents from potentials positive to -80 mV was fitted by a Boltzmann equation revealing the existence of an inactivating transient outward component (I(to)) and a noninactivating steady-state component (I(ss)). ET-1 markedly inhibited I(ss) by 43.0 +/- 3.8% (P < 0.001, n = 7) and shifted the voltage dependent inactivation of I(to) by +3.3 +/- 1.2 mV (P < 0.05). Although ET-1 had little effect on the onset of inactivation of the currents elicited from a conditioning potential of -70 mV, the time-independent noninactivating component of the currents was markedly inhibited. In conclusion, the predominant effect of ET-1 was to inhibit a noninactivating steady-state background K(+) current (I(ss)). These results are consistent with the hypothesis that I(ss) inhibition contributes to the inotropic effects of ET-1. PMID- 11409901 TI - Cloning of full-length genomic DNA encoding human FcepsilonRI alpha-chain and its transcriptional regulation. AB - Two novel exons, named exon 1A and exon 2A, were found at 18.4 and 12.6 kb upstream from the exon known as the first exon of human FcepsilonRI alpha-chain gene. Transcription from the promoter present in the upstream of exon 1A was decreased by mutations introduced into the "first intron" between exon 1A and exon 2A, suggesting the presence of an intronic regulatory element in the intron. Consistent with this, electrophoretic mobility shift assay revealed the presence of a nuclear factor which bound the region in FcepsilonRI alpha-chain positive cells. PMID- 11409902 TI - Cloning of the human cholesteryl ester hydrolase promoter: identification of functional peroxisomal proliferator-activated receptor responsive elements. AB - Cholesteryl ester hydrolase (CEH) is responsible for hydrolysis of stored cholesterol esters in macrophage foam cells and release of free cholesterol for high-density lipoprotein-mediated efflux. PCR-based screening of human genomic libraries with human macrophage CEH specific primers resulted in amplification and cloning of 1.7 kb promoter sequence. Analysis of the sequence revealed a lack of consensus TATA-box but presence of a GC-rich proximal sequence, a CAAT box and several binding sites for the transcription factor Sp1. Three putative response elements for peroxisome proliferator-activated receptor (PPRE) were identified at position -176, -779, and -1316. Down-regulation of promoter activity was observed in the presence of either PPARalpha- or PPARgamma-specific ligands and introduction of a 4-point transverse mutation in the PPRE at -176 completely abolished the effect of PPAR ligands on the promoter activity. Analogous to other genes involved in macrophage cholesterol homeostasis, human CEH may also be regulated by PPAR. PMID- 11409903 TI - Melatonin activates the peroxidase-oxidase reaction and promotes oscillations. AB - We have studied the peroxidase-oxidase reaction with NADH and O2 as substrates and melatonin as a cofactor in a semibatch reactor. We show for the first time that melatonin is an activator of the reaction catalyzed by enzymes from both plant and animal sources. Furthermore, melatonin promotes oscillatory dynamics in the pH range from 5 to 6. The frequency of the oscillations depends on the pH such that an increase in pH was accompanied by a decrease in frequency. Conversely, an increase in the flow rate of NADH or an increase in the average concentration of NADH resulted in an increase in oscillation frequency. Complex dynamics were not observed with melatonin as a cofactor. These results are discussed in relation to observations of oscillatory dynamics and the function of melatonin and peroxidase in activated neutrophils. PMID- 11409904 TI - Suppression of platelet-type 12-lipoxygenase activity in human erythroleukemia cells by an RNA-cleaving DNAzyme. AB - Human platelet-type 12-lipoxygenase (12-LOX) and its metabolites play a crucial role in tumor angiogenesis. A "10-23" deoxyribozyme (DNAzyme) and its phosphorothioate-modified version were designed and synthesized against the 12 LOX mRNA. Both DNAzymes were able to cleave their substrate efficiently in a time and concentration-dependent manner in vitro. Under a multiple turnover condition, both performed well at 37 degrees C, showing the k(cat) of 1 and 0.26 min(-1), respectively. The phosphorothioate modification of the DNAzyme significantly increased its stability in cells without a substantial loss of kinetic efficiency in vitro. In a cell culture system, transfection of the DNAzymes into HEL cells resulted in a significant down-regulation of the 12-LOX mRNA. Furthermore, the cell extracts from the DNAzyme-transfected cells exhibited a marked reduction in the 12-LOX enzyme activity. The present results indicated the potential use of DNAzyme technology for gene function study and cancer therapy. PMID- 11409905 TI - Similar subunit interactions contribute to assembly of clathrin adaptor complexes and COPI complex: analysis using yeast three-hybrid system. AB - Clathrin adaptor protein (AP) complexes are heterotetramers composed of two large, one medium, and one small subunits. By exploiting the yeast three-hybrid system, we have found that an interaction between the two large subunits of the AP-1 complex, gamma-adaptin and beta1-adaptin, is markedly enhanced in the presence of the small subunit, sigma1. Similarly, two large subunits of the AP-4 complex, epsilon-adaptin and beta4-adaptin, are found to interact with each other only in the presence of the small subunit, sigma4. Furthermore, we have found that an interaction between two large subunits of the COPI F subcomplex, gamma COP and beta-COP, is detectable only in the presence of zeta-COP. Because these COPI subunits have common ancestral origins to the corresponding AP subunits, these three-hybrid data, taken together with the previous two-hybrid data, suggest that the AP complexes and the COPI F subcomplex assemble by virtue of similar subunit interactions. PMID- 11409906 TI - Cancer and thrombosis revisited. AB - Remarkable progress has been made since the first description of the association between cancer and thrombosis by Trousseau over 100 years ago. Now, it is clear that there is a two-way connection between coagulation and cancer as tumor results in alterations in hemostatic balance, and thrombosis may promote tumor cell growth. A variety of clinical thrombotic syndromes may present in cancer patients including local and systemic venous and arterial thromboses. More evidence is now being gathered on the potential of antithrombotic regimens to prolong survival of cancer patients. Whether the use of novel antithrombotic drugs may result in a better outcome remains to be determined. PMID- 11409907 TI - Non-infectious complications of transfusion therapy. AB - Blood transfusion is considered safe when the infused blood is tested using state of the art viral assays developed over the past several decades. Only rarely are known viruses like HIV and hepatitis C transmitted by transfusion when blood donors are screened using these sensitive laboratory tests. However, there are a variety of transfusion risks which still remain that cannot be entirely eliminated, many of which are non-infectious in nature. Predominantly immune mediated complications include the rapid intravascular or slow extravascular destruction (hemolysis) of transfused red cells or extravascular removal of platelets by pre-formed antibodies carried by the transfusion recipient. Alternatively, red cells can be damaged when exposed to excessive heat or incompatible intravenous fluids before or during the transfusion. Common complications of blood transfusion that at least partly involve the immune system include febrile non-hemolytic and allergic reactions. While these are usually not life-threatening, they can hamper efforts to transfuse a patient. Other complications include circulatory overload, hypothermia and metabolic disturbances. Profound hypotensive episodes have been described in patients on angiotensin-converting enzyme (ACE) inhibitors who receive platelet transfusions through bedside leukoreduction filters. These curious reactions appear to involve dysmetabolism of the vasoactive substance bradykinin. Products contaminated by bacteria during blood collection and transfused can cause life-threatening septic reactions. A long-term complication of blood transfusion therapy unique to chronically transfused patients is iron overload. Less common - but serious - reactions more specific to blood transfusion include transfusion-associated graft versus-host disease and transfusion-associated acute lung injury. Many of these complications of transfusion therapy can be prevented by adhering to well established practice guidelines. In addition, individuals who administer blood transfusions should recognize these complications in order to be able to quickly provide appropriate treatment. PMID- 11409908 TI - Tyrosine kinase inhibitors in the treatment of chronic myeloid leukaemia: so far so good? AB - Chronic myeloid leukaemia (CML) is characterized by marked expansion of the myeloid series, and is thought to arise as a direct result of the bcr-abl fusion gene. The BCR-ABL oncoprotein is a constitutively active protein tyrosine kinase (PTK), which results in altered cell signalling and is responsible for the changes that characterize the malignant cells of CML. It has been shown that the increased tyrosine kinase activity of BCR-ABL is a requirement for transformation and is, therefore, a legitimate target for pharmacological inhibition. Several compounds have now been identified as relatively selective inhibitors of BCR-ABL, including members of the tyrphostin family, herbimycin A and most importantly the 2-phenylaminopyrimidine ST1571. Having established the efficacy of this agent in vitro, phase I trials using an oral formulation were commenced in the USA in mid 1998. Early data from an interferon-alpha (IFN) resistant/refractory or intolerant cohort demonstrated good patient tolerance and effective haematological control at doses above 300 mg. More promising was its ability to induce cytogenetic responses in this pretreated group of patients. Phase II data, albeit far from complete, appear to confirm its efficacy even in the context of advanced disease and phase III clinical trials are currently underway in many countries. Recent laboratory evidence, however, suggests that the development of drug resistance is a possibility (via amplification of the bcr-abl fusion gene, overexpression of P-glycoprotein or binding of ST1571 to alpha1 acid glycoprotein) and that combination therapy including ST1571 should be considered. PMID- 11409909 TI - Laboratory correlates in multiple myeloma: how useful for prognosis? PMID- 11409910 TI - Extracorporeal photopheresis: a review. AB - Extracorporeal phototherapy (ECP) is a therapeutic approach based on the biological effect of psoralen 8-methoxypsoralen (8-MOP) and ultraviolet light A (UVA) on mononuclear cells collected by apheresis, and reinfused into the patient. Photopheresis is widely used for the treatment of patients with advanced cutaneous T-cell lymphoma (CTLC). Evidence suggests that it prolongs life, and also induces 50-75% response rates. In addition, more and more reports indicate that photopheresis is a potent agent in the therapy of acute allograft rejection among cardiac, lung and renal transplant recipients. There are increasing amounts of data showing that patients with chronic graft versus host disease benefit from this therapy. Likewise, there are indications that there may be a potential role for ECP in the therapy of certain autoimmune diseases resistant to conventional therapy. The mechanism of this treatment is likely due to the induction of anticlonotypic immunity directed against pathogenic clones of T lymphocytes. Treatment induces apoptotic death of pathogenic T-cells, and it is postulated that activation of antigen-presenting cells has important effects in this process. PMID- 11409911 TI - Genetic and molecular mechanisms of cell size control. AB - Since the discovery of the cell as the minimal indivisible living entity, scientists have tried to understand how all the various physiological aspects were assumed within this single functional unit. One fascinating question concerns the role of cell size and cell number in determining the overall size of an organism. During the past century, increasing knowledge in molecular genetics has allowed the characterization of a number of molecular events that influence the size of a cell. However, in spite of recent progress, precise molecular mechanisms governing cell size remain unclear. Although the existence of a master regulator is still possible, cell size may be primarily controlled by an interactive network linking gene expression with translational capacity and cell proliferation. PMID- 11409912 TI - Exogenous fibroblast growth factor-2 induces a transformed phenotype in vascular kaposi's sarcoma-like cells. AB - Vascular TTB cells derive from murine Kaposi's sarcoma-like dermal lesions and share several phenotypic features with AIDS-associated KS spindle cells. We have recently reported that fibroblast growth factor-2 (FGF-2) promotes dramatic cytoskeletal and morphological alterations in TTB cells, concomitant with the induction of an autocrine loop for hepatocyte growth factor and a relocalization of the urokinase receptor. Since all these alterations are hallmarks of cell transformation. we attempted to verify whether FGF-2 induces a transformed phenotype in TTB cells. Our results show that FGF-2-treated TTB cells acquire the ability to grow under anchorage-independent conditions. In addition, FGF-2 markedly reduced the levels of thrombospondin-1, an antiangiogenic and tumor suppressor protein, in TTB cells. Therefore, FGF-2 induces KS-like spindle cells to acquire properties characteristic of transformed cells. This suggests that FGF 2 plays a pathogenetic role in KS not only by promoting angiogenesis, but also by conferring a transformed phenotype upon KS cells. In light of previous reports on Tat-induced release of FGF-2 into the extracellular space, our findings may provide an additional mechanism for the observed synergism between Tat and FGF-2 in the pathogenesis of KS. PMID- 11409913 TI - An open chromatin structure in a liver-specific enhancer that confers high level expression to human apolipoprotein b transgenes in mice. AB - A number of DNaseI-hypersensitive (DH) sites have been mapped within a regulatory region situated upstream of the human apolipoprotein B (apoB) promoter (-5262 to 899) that is required for high level expression of human apoB transgenes in the livers of mice. These DH sites were observed in nuclei from transcriptionally active liver-derived HepG2 cells, but were absent from transcriptionally inactive HeLa cell nuclei. Several nuclear protein binding sites were detected in the DNaseI-hypersensitive region by DNaseI footprinting with HepG2 nuclear extracts, representing putative binding sites for the liver-specific activators. The locations of binding sites for these transcription factors were revealed via computer analysis of the DNA sequence of this region against a transcription factor database. Many micrococcal nuclease hypersensitive (MH) sites were also observed in nuclei from HepG2 cells but not in HeLa cell nuclei, implying that in hepatic cells, nucleosomes are either absent or have been displaced from this region by the liver-specific transcriptional activators, as inferred by the correspondence between the DH sites, the MH sites and the footprints. PMID- 11409914 TI - Glutathione transport system in NIH3t3 fibroblasts. AB - The current study characterizes a mediated transport for GSH uptake in murine fibroblasts NIH3T3. The presence of GSH mediated transport is indicated by the behaviour of GSH uptake time-course, as well as by kinetic saturation and the specific inhibition of the initial rate of GSH transport. Moreover, a concentrative GSH uptake has been measured, whose driving force may be due to a change of membrane potential and the direct involvement of ATP. Hyperbolic kinetic saturation shows a single mediated transport with high affinity for GSH (Km = 0.209 +/- 0.03 mM). High specificity of this GSH transporter for the entire structure of GSH is also demonstrated. To summarize, GSH uptake into NIH3T3 cells occurs by an active transport system and shows characteristics similar to ATP dependent mechanisms previously identified in hepatocyte membranes. Moreover, a possible physiological role of this GSH transporter related to NIH3T3 cell proliferation has been hypothesized. PMID- 11409915 TI - BAP37 and Prohibitin are specifically recognized by an SV40 T antigen antibody. AB - We have identified two cellular proteins that are specifically immunoprecipitated by an anti-SV40 T antigen monoclonal antibody. This antibody, PAb419, recognizes an epitope contained within a region of T antigen which we have recently demonstrated is required for the initiation of immortalization by SV40 T antigen, but is not essential for maintenance of the immortal state. The two proteins were identified as BAP37 and Prohibitin. Recent results suggest Prohibitin may enhance the transcriptional inactivation of E2F by the retinoblastoma family of pocket proteins (pRb, p107, p130). BAP37 and Prohibitin are specifically recognized by PAb419 and PAb210, another anti-SV40 T antigen monoclonal antibody, which has an overlapping epitope, but not by other anti-SV40 T antigen monoclonal antibodies, demonstrating the specificity of the interaction. PMID- 11409916 TI - Oligomeric status of the dihydropyridine receptor in aged skeletal muscle. AB - A prominent feature of aging is represented by a decrease in muscle mass and strength. Abnormalities in Ca2+ -regulatory membrane complexes are involved in many muscular disorders. In analogy, we determined potential age-related changes in a key component of excitation-contraction coupling, the dihydropyridine receptor. Immunoblotting of the microsomal fraction from aged rabbit muscle revealed a drastic decline in the voltage-sensing alpha1-subunit of this transverse-tubular receptor, but only marginally altered expression of its auxiliary alpha(2)-subunit and the Na+/K+ -ATPase. A shift to slower fibre type characteristics was indicated by an age-related increase in the slow calsequestrin isoform. Chemical crosslinking analysis showed that the triad receptor complex has a comparable tendency of protein-protein interactions in young and aged muscles. Hence, a reduced expression and not modified oligomerization of the principal dihydropyridine receptor subunit might be involved in triggering impaired triadic signal transduction and abnormal Ca2+ homeostasis resulting in a progressive functional decline of skeletal muscles. PMID- 11409917 TI - Support vector machines for prediction of protein subcellular location. AB - Support Vector Machine (SVM), which is one kind of learning machines, was applied to predict the subcellular location of proteins from their amino acid composition. In this research, the proteins are classified into the following 12 groups: (1) chloroplast, (2) cytoplasm, (3) cytoskeleton, (4) endoplasmic reticulum, (5) extracall, (6) Golgi apparatus, (7) lysosome, (8) mitochondria, (9) nucleus, (10) peroxisome, (11) plasma membrane, and (12) vacuole, which have covered almost all the organelles and subcellular compartments in an animal or plant cell. The examination for the self-consistency and the jackknife test of the SVMs method was tested for the three sets: 2022 proteins, 2161 proteins, and 2319 proteins. As a result, the correct rate of self-consistency and jackknife test reaches 91 and 82% for 2022 proteins, 89 and 75% for 2161 proteins, and 85 and 73% for 2319 proteins, respectively. Furthermore, the predicting rate was tested by the three independent testing datasets containing 2240 proteins, 2513 proteins, and 2591 proteins. The correct prediction rates reach 82, 75, and 73% for 2240 proteins, 2513 proteins, and 2591 proteins, respectively. PMID- 11409918 TI - Complexes formation between insulin receptor and extracellular signal-regulated kinases ERKs. AB - A property of signal transduction pathways that might explain their efficiency and specificity is the formation of signaling complexes. The recent demonstration that adaptor proteins can interact with many components of the extracellular signal-regulated kinases (ERKs) signaling cascade leads us to investigate whether such complexes may include the transmembrane receptor. The present work shows that in human hepatoma Hep3B cells, insulin receptor (IR) can be coimmunoprecipitated with other components of the ERKs cascade: insulin receptor substrate (IRS), Raf-1, and ERKs. Furthermore, these complexes formed near the cytoplasmic membrane even prior to insulin stimulation. PMID- 11409919 TI - In vivo ribozyme targeting of betaAPP+ mRNAs. AB - In Alzheimer's disease (AD) and Down's syndrome (DS) patients, posttranscriptional alterations of sequences encoded by exon 9 and exon 10 of the beta-amyloid precursor protein (betaAPP) mRNA result in mutant proteins (betaAPP+) that colocalize with neurofibrillary tangles and senile plaques. These aberrant messages may contribute to the development of sporadic or late-onset Alzheimer's disease; thus, eliminating them or attenuating their expression could significantly benefit AD patients. In the present work, self-cleaving hammerhead ribozymes targeted to betaAPP exon 9 (Rz9) and betaAPP+ mutant exon 10 (Rz10) were examined for their ability to distinguish between betaAPP and betaAPP+ mRNA. In transiently transfected A-204 cells, quantitative confocal fluorescence microscopy showed that Rz9 preferentially lowered endogenous betaAPP. In contrast, in transient cotransfection experiments with betaAPP+ mRNAs containing a wild-type exon 9 and mutant exon 10 (betaAPP-9/betaAPP-10+1), or a mutant exon 9 and wild-type exon 10 (betaAPP-9+1/betaAPP-10) we found that Rz9 and Rz10 preferentially reduced betaAPP+ -mutant exon 10 mRNA in a concentration and a ribozyme-dependent manner. PMID- 11409920 TI - Inflammation and interleukin-6 mediate reductions in the hepatic expression and transcription of the mdr1a and mdr1b Genes. AB - In order to elucidate the mechanisms involved in the downregulation of mdr 1 gene expression reported in experimentally-induced inflammation, we examined the effects of experimentally-induced inflammation and interleukin-(IL-) 6 on transcriptional control of the mdr1 genes in rats. RNA, nuclear extracts, and nuclear protein fractions were isolated from livers harvested from saline or turpentine-treated male Sprague-Dawley rats or from IL-6 treated or nontreated (controls) cultured rat hepatocytes. mdr gene expression and regulation was examined by RT-PCR, mRNA stability studies, nuclear run-on analysis of transcription, and gel shift analysis of promoter-transcription factor interaction. As compared to controls, significantly lower levels of mdr1a and mdr1b mRNA and significantly decreased mdr1a and mdr1b transcription rates were observed in livers isolated from the turpentine-treated rats. In vitro treatments of cultured hepatocytes with IL-6 also suppressed mdr1a and mdr1b mRNA expression and imposed similar reductions in mdr1a and mdr1b transcriptional activity. Significant effects of IL-6 on mdr1 mRNA stability were not seen. Our results indicate that reductions in mdr1 expression in experimental models of inflammation likely occurs through altered gene transcription. Furthermore, as IL 6 was found to decrease mdr1 expression and gene transcription rates in vitro, this cytokine is likely involved in the reduction of mdr1 expression that is seen in vivo during an acute inflammatory response. PMID- 11409921 TI - The virtual patient -- addressing the placebo challenge. PMID- 11409922 TI - The Ross Report: implications for veterinary education. PMID- 11409923 TI - Pharmacological treatment in behavioural medicine. PMID- 11409924 TI - The effect of forage type on foot health in dairy heifers. PMID- 11409925 TI - Pharmacological treatment in behavioural medicine: the importance of neurochemistry, molecular biology and mechanistic hypotheses. AB - Psychopharmacology has become a popular, and sometimes mandatory addition to treatment regimes for canine and feline patients with behavioural problems; however, clients and practitioners should be dissuaded that behavioural drugs are 'quick fixes'. Veterinarians should only prescribe psychotropic medication when they have a specific idea of how the mechanism of action of the drug will affect the target behaviours associated with a specific diagnosis. The diagnosis must be treated rather than non-specific signs. Newer psychotropic medications demonstrate the extent to which truly abnormal behaviours are dysfunctions of neurochemistry; synaptic or cellular metabolism; or genetic encoding and 'learning', or LTP, hence there is a clear role for the interaction of neuropharmacology and behavioural and environmental modification.Future advances in treatment in behavioural medicine will be pharmacological and neurophysiological. As the field of behavioural medicine expands, its paradigm will enlarge to include combination therapy and the implementation of neuropharmacological intervention as a diagnostic tool. At present, the veterinary practitioner can effectively aid many common behavioural problems using extant drugs to treat animals with true behavioural pathology. Rational pharmacological therapy requires complete medical and behavioural histories, requisite laboratory work, complete client understanding and compliance, and an honest and ongoing dialogue between the client and veterinarian that includes frequent follow-ups and re-examinations. PMID- 11409926 TI - Training affects the collagen framework of subchondral bone in foals. AB - Subchondral bone provides structural support to the overlying articular cartilage and plays an important role in osteochondral diseases. There is growing insight that the mechanical features of bone are related to the biochemistry of the collagen network. In this study the effect of exercise on water, calcium and the collagen network (total collagen, lysyl-hydroxylation, hydroxylysylpyridinoline, and lysylpyridinoline crosslinks) of subchondral bone at two differently loaded sites (site 1: intermittently loaded; site 2: constantly loaded) is investigated in foals. Exercise influenced calcium content and levels of both types of crosslinks at site 1, but had no influence on site 2. There was no concomitant increase in lysyl-hydroxylation level with the rise in crosslinks. Levels of lysyl-hydroxylation and lysylpyridinoline crosslinking were lower at site 1 than at site 2. It is concluded that exercise affects the post-translational modifications of the collagen component of subchondral bone. Loading also appears to play a role in site-related topographical differences. The lack of any relation between the sum of pyridinoline crosslinks and the amount of triple helical hydroxylysine gives support to a recent hypothesis that lysyl hydroxylation of the triple helix and the telopeptides are under separate control. PMID- 11409927 TI - Strangulation of the duodenum by the uterus during late pregnancy in two cows. AB - Two Swiss Braunvieh cows in late pregnancy underwent surgery because of a rare form of ileus due to strangulation of the duodenum at its caudal flexure by the gravid uterus. The whole uterus had passed through a gap between the mesoduodenum and duodenum and with increasing weight had led to strangulation of the duodenum. This was possible since the mesoduodenum and both walls of the greater omentum adjacent to its caudal edge were not connected with the duodenum, probably due to a congenital inhibitory malformation. A transsection and an end-to-end anastomosis of the duodenum were necessary in both cases since it was impossible to retract the gravid uterus through the defect. Postoperative recovering was uneventful in both cows, which were discharged after seven and five days respectively and calved normally about two months later. PMID- 11409928 TI - Disposition of suprofen enantiomers in the cat. AB - Suprofen (SPF) is a non-steroidal anti-inflammatory drug (NSAID), which belongs to the 2-arylpropionic acids subclass. As a result of their chiral characteristics, these compounds have shown a marked enantioselective behaviour with a high degree of interspecies variation. They are mainly eliminated by glucuronidation. Plasma, biliary and urine disposition of SPF was investigated in the cat after intravenous administration of the racemate (dose 2 mg/kg). Both enantiomers exhibited similar disposition profiles in plasma with no evidence of chiral inversion. During bile sampling time, recovered acylglucuronides of R (-) and S (+) SPF were less than 1% of the total dose administered. Only free SPF was recovered in the urine, representing 0.12% of the administered racemic SPF dose. The results indicate that neither chiral inversion nor glucuronidation predominate in SPF disposition in cats. PMID- 11409929 TI - Cardiovascular, haematological and biochemical responses after large volume blood collection in horses. AB - To determine whether removal of 20 mL/kg of blood (approximately 25% of blood volume) resulted in adverse physiological effects in donor horses, we removed this volume of blood from five horses and selected cardiovascular, haematological and biochemical variables measured during collection and for 31 days thereafter. We found that alteration in most variables occurred, although the changes in values usually remained within published reference ranges. Also, recovery of these alterations to pre-collection values was rapid, occurring within 24--48 h in most instances. We concluded that volumes of blood less than or equal to 20 mL/kg when collected appropriately from healthy donor horses result in no adverse acute or chronic physiological changes. These results suggest that horses undergo adequate physiological compensation when approximately 25% of blood volume is removed for the purposes of blood donation or production of plasma. PMID- 11409930 TI - Effects of housing and two forage diets on the development of claw horn lesions in dairy cows at first calving and in first lactation. AB - This paper describes a systematic study of the development of lesions of the claw horn (CHL, sole and white line) in heifers calving for the first time, housed either in cubicles or a straw yard and fed either a low- or high-dry-matter forage diet. The feet of all animals were inspected on five occasions, at approximately four weeks before and four, eight, 16 and 24 weeks post calving. Haemorrhagic lesions of the sole and white line were described according to a geometric lesion score for severity and a cumulative lesion score based on the product of (severity x area) for each lesion. Geometric and cumulative lesion scores increased in all groups of cattle in the first eight weeks after calving. However, the severity and persistence of the lesions were significantly greater in cattle housed in cubicle yards. Wet feeding increased the severity of CHL in the cubicle yard only. There were no associations between lesion scores and body weight, body condition or foot conformation. The heels of the cattle in straw yards tended to be thick but many showed pitting erosions. In cubicles the heels were smooth but thin. This may have contributed to CHL by increasing concussive forces within the hoof. There was a highly significant (but relatively low) correlation between scores for sole lesions and lameness in individual animals. These observations are consistent with the hypothesis that systemic events associated with calving and the onset of lactation may set in motion the chain of events that lead to the lesions of CHL; the extent and severity of these lesions being then determined by the externally imposed conditions of housing and feeding. PMID- 11409931 TI - Antibodies to selected canine pathogens and infestation with intestinal helminths in golden jackals (Canis aureus) in Israel. AB - Blood and fecal samples, collected from 46 healthy adult free-ranging golden jackals captured in two different locations in Israel, were examined. A serological Study was conducted to investigate the prevalence of circulating antibodies reacting with four common canine pathogens: canine distemper virus (CDV), canine parvovirus (CPV), Ehrlichia canis and Leishmania infantum. Faecal flotation and haematological tests were also performed. The seroprevalence of CPV, E. canis, CDV, and L. infantum were 72.3% (34/47), 54.3% (25/46), 52.2% (24/46), and 6.5% (3/46) respectively. Faecal flotation tests revealed a high prevalence of Ancylostoma caninum (13/17, 76%) and a low prevalence of Dipilidium caninum infestation. Examination of blood smears revealed Hepatazoon canis gamonts in one jackal. Golden jackals are among the most common free-ranging carnivores in Israel and neighboring countries. Their habitats are in proximity to densely populated areas and they bear close phylogenic relation to the domestic dog. These facts, combined with the high prevalence of the jackals' exposure to the major canine pathogens demonstrated in this study, suggest that they may serve as a reservoir for the transmission of certain diseases to domestic dogs. PMID- 11409932 TI - The effect of sodium heparin on equine articular cartilage. AB - This study compared the effect of sodium heparin and gentamicin sulphate on equine articular cartilage (AC) explants in order to investigate the possible use of sodium heparin in the treatment of infectious arthritis. Six concentrations of sodium heparin and gentamicin sulphate were tested. The supernatant and explant digest were assayed for glycosaminoglycan (GAG) content with the dimethyl methylene blue assay and the per cent loss of GAG was calculated. A significant (P< 0.001) increase in percentage GAG loss was noted for the sodium heparin groups when compared to the control, whilst no significant increase was found among the treatment groups (P =0.782). For gentamicin, no significant difference in percentage GAG loss was found between the control and three of the five treatment groups (P =0.667). The percentage GAG loss in the sodium heparin treated AC explants was greater than for any of the gentamicin-treated AC explants. It can be concluded that sodium heparin sulphate stimulates an increase in GAG release from equine articular cartilage explants, though no firm conclusions can be drawn on its use in treating equine infectious arthritis. PMID- 11409933 TI - Common and uncommon cytochrome P450 reactions related to metabolism and chemical toxicity. AB - Cytochrome P450 (P450) enzymes catalyze a variety of reactions and convert chemicals to potentially reactive products as well as make compounds less toxic. Most of the P450 reactions are oxidations. The majority of these can be rationalized in the context of an FeO(3+) intermediate and odd electron abstraction/rebound mechanisms; however, other iron-oxygen complexes are possible and alternate chemistries can be considered. Another issue regarding P450 catalyzed reactions is the delineation of rate-limiting steps in the catalytic cycle and the contribution to reaction selectivity. In addition to the rather classical oxidations, P450s also catalyze less generally discussed reactions including reduction, desaturation, ester cleavage, ring expansion, ring formation, aldehyde scission, dehydration, ipso attack, one-electron oxidation, coupling reactions, rearrangement of fatty acid and prostaglandin hydroperoxides, and phospholipase activity. Most of these reactions are rationalized in the context of high-valent iron-oxygen intermediates and Fe(2+) reductions, but others are not and may involve acid-base catalysis. Some of these transformations are involved in the bioactivation and detoxication of xenobiotic chemicals. PMID- 11409934 TI - Monomethylarsonous acid (MMA(III)) and arsenite: LD(50) in hamsters and in vitro inhibition of pyruvate dehydrogenase. AB - Monomethylarsonous acid (MMA(III)), a metabolite of inorganic arsenic, has received very little attention from investigators of arsenic metabolism in humans. MMA(III), like sodium arsenite, contains arsenic in the +3 oxidation state. Although we have previously demonstrated that it is more toxic than arsenite in cultured Chang human hepatocytes, there are no data showing in vivo toxicity of MMA(III). When MMA(III) or sodium arsenite was administered intraperitoneally to hamsters, the LD(50)s were 29.3 and 112.0 micromol/kg of body wt, respectively. In addition, inhibition of hamster kidney or purified porcine heart pyruvate dehydrogenase (PDH) activity by MMA(III) or arsenite was determined. To inhibit hamster kidney PDH activity by 50%, the concentrations (mean +/- SE) of MMA(III) as methylarsine oxide, MMA(III) as diiodomethylarsine, and arsenite were 59.9 +/- 6.5, 62.0 +/- 1.8, and 115.7 +/- 2.3 microM, respectively. To inhibit activity of purified porcine heart PDH activity by 50%, the concentrations (mean +/- SE) of MMA(III) as methylarsine oxide and arsenite were 17.6 +/- 4.1 and 106.1 +/- 19.8 microM, respectively. These data demonstrate that MMA(III) is more toxic than inorganic arsenite, both in vivo and in vitro, and call into question the hypothesis that methylation of inorganic arsenic is a detoxication process. PMID- 11409935 TI - Gibbs energy of formation of peroxynitrite--order restored. AB - In a recent publication [Nauser et al. (2001) Chem. Res. Toxicol. 14, 248-350], the authors estimated a value of 14 +/- 3 kcal/mol for the standard Gibbs energy of formation of ONOO(-) and argued that the experimental value of 16.6 kcal/mol [Merenyi, G., and Lind, J. (1998) Chem. Res. Toxicol. 11, 243-246] is in error. The lower value would suggest that the yield of free radicals during decomposition of ONOOH into nitrate is negligibly low, i.e., less than 0.5%, though within the large error limit given, the radical yield might vary between 0.003% and ca. 80%. The experimental value of 16.6 +/- 0.4 kcal/mol was based on the determination of the rate constant of the forward reaction in the equilibrium ONOO(-) <==> (*)NO and O2(*-) by use of C(NO2)4, an efficient scavenger of O2(*-) which yields C(NO2)3(-). Nauser et al. reported that addition of (*)NO has no significant effect on the rate of formation of C(NO2)3(-), and therefore the formation of C(NO2)3(-) is due to a process other then reduction of C(NO2)4 by O2(*-). In addition, they argued that Cu(II) nitrilotriacetate enhances the rate of peroxynitrite decomposition at pH 9.3 without reduction of Cu(II). In the present paper, we show that the formation of C(NO2)3(-) due to the presence peroxynitrite is completely blocked upon addition of (*)NO. Furthermore, the acceleration of the rate of peroxynitrite decomposition at pH 9 in the presence of catalytic concentrations of SOD ([ONOO(-)]/[SOD] > 30) results in the same rate constant as that obtained in the presence of C(NO(2))(4). These results can only be rationalized by assuming that ONOO(-) homolyses into (*)NO and O2(*-) with k = 0.02 s(-1) at 25 degrees C. Thus, the critical experiments suggested by Nauser et al. fully support the currently accepted thermodynamics as well as the mode of decomposition of the ONOOH/ONOO(-) system. PMID- 11409936 TI - An evaluation of the mutagenicity, metabolism, and DNA adduct formation of 5 nitrobenzo[b]naphtho[2,1-d]thiophene. AB - Thioarenes, sulfur-containing polycyclic aromatic compounds, are environmental contaminants suspected of posing human health risks. In this study, 5 nitrobenzo[b]naphtho[2,1-d]thiophene (5-nitro-BNT), a nitrated-thioarene, was examined for its mutagenicity, metabolism and subsequent formation of DNA adducts. 5-Nitro-BNT was weakly mutagenic in Salmonella typhimurium strains TA98 and TA100 without Aroclor-1254-induced rat liver S9 (S9), and its activity was increased in the presence of S9. Anaerobic metabolism of 5-nitro-BNT by S9 or xanthine oxidase (XO) produced one major metabolite, identified as 5-amino-BNT by NMR, MS, and UV spectroscopy and by comparison with an authentic standard. Aerobic S9 metabolism of 5-nitro-BNT produced a major metabolite, identified as trans-9,10-dihydroxy-9,10-dihydro-5-nitro-BNT (5-nitro-BNT-9,10-diol). Also present was a minor amount of 5-amino-BNT and trans-9,10-dihydroxy-9,10-dihydro-5 amino-BNT (5-amino-BNT-9,10-diol). DNA adduct analyses were performed using the (32)P-postlabeling assay and reversed-phase HPLC. Three major XO-derived calf thymus DNA adducts were detected. On the basis of their chromatographic mobilities, two adducts were identified as reaction products of 5-nitro-BNT with 2'-deoxyguanosine and one adduct with 2'-deoxyadenosine. Incorporation of allopurinol (a specific XO inhibitor) in the incubation mixture resulted in loss of all three adducts, confirming enzymatic mediation by XO. Aerobic S9 activation of 5-nitro-BNT with calf thymus DNA produced three adducts. On the basis of their chromatographic mobilities, two were identified as reaction products of 5-nitro BNT with 2'-deoxyguanosine and one with 2'-deoxyadenosine. Incorporation of 1 aminobenzotriazole (a P450 inhibitor) in the incubation mixture resulted in a loss of these adducts, confirming enzymatic mediation by P450. Aerobic S9 catalyzed metabolism of 5-nitro-BNT-9,10-diol produced the same DNA adducts as observed with 5-nitro-BNT. Aerobic S9-catalyzed metabolism of 5-amino-BNT-9,10 diol produced the same deoxyadenosine-derived DNA adducts as observed with 5 nitro-BNT and 5-nitro-BNT-9,10-diol. These results provide additional information that both ring oxidation and nitroreduction are involved in the metabolism, DNA adduct formation and mutagenicity of 5-nitro-BNT. PMID- 11409937 TI - Oxidation of 2,6-dimethylaniline by recombinant human cytochrome P450s and human liver microsomes. AB - 2,6-Dimethylaniline (2,6-DMA) is classified as a rodent nasal cavity carcinogen and a possible human carcinogen. The major metabolite of 2,6-DMA in rats and dogs is 4-amino-3,5-dimethylphenol (DMAP) but oxidization of the amino group to produce metabolites such as N-(2,6-dimethylphenyl)hydroxylamine (DMHA) is also indicated by the occurrence of hemoglobin adducts of 2,6-DMA in human and rats. Previous studies have shown a large interindividual variability in human 2,6-DMA hemoglobin adduct levels. In the present study, 2,6-DMA oxidation in vitro by human liver microsomes and recombinant human P450 enzymes was investigated to assess whether the hemoglobin adduct variability could be attributed to metabolic differences. At micromolar concentrations, the only product detectable (UV) was DMAP, while at 10 nM, DMHA was a substantial product. 2E1 and 2A6 were identified as the major P450s in human liver microsomes responsible for the production of DMAP by using P450-specific chemical inhibitors and mouse monoclonal antibodies that selectively inhibit human P450 2E1 and 2A6. 2A6 was identified as the major P450 responsible for the N-hydroxylation. Native P450 2E1 and human liver microsomes catalyzed the rearrangement of DMHA to DMAP independent of NADPH. Consistent with a mechanism involving oxygen rebound to the heme iron center, labeled oxygen was not incorporated into DMAP from either 18O2 gas or H2 18O in this rearrangement. Results presented here suggest much of the observed interindividual variability of 2,6-DMA hemoglobin adduct levels could be due to differences in the relative amounts of hepatic 2E1 and 2A6. PMID- 11409938 TI - Requirement of glutathione and cysteine in guanine-specific oxidation of DNA by carcinogenic potassium bromate. AB - Potassium bromate (KBrO3), a food additive, induces renal-cell tumors in rats. KBrO3 induced 8-oxo-7, 8-dihydro-2'-deoxyguanosine (8-oxodG) formation in human leukemia cell line HL-60 as well as in its H2O2-resistant clone, HP100, suggesting no involvement of H2O2. Depletion of GSH by buthionine sulfoximine (BSO) had a little inhibitory effect on KBrO3-induced 8-oxodG formation. However, the amount of 8-oxodG was still significantly higher than that in control, suggesting that intracellular Cys can affect KBrO3 to oxidize DNA, when GSH decreased. KBrO3 caused 8-oxodG in isolated DNA in the presence of GSH (tripeptide; gamma-GluCysGly), gamma-GluCys, CysGly, or Cys. Methional completely inhibited 8-oxodG formation induced by KBrO3 plus GSH, but typical hydroxyl radical scavengers, SOD and catalase, had little or no inhibitory effects. When bromine solution (BrO(-)) was used instead of BrO3(-), similar scavenger effects were observed. Experiments with 32P-labeled DNA fragments obtained from the human p53 tumor suppressor gene and the c-Ha-ras-1 protooncogene suggested that KBrO3 induced 8-oxodG formation at 5'-site guanine of GG and GGG sequences of double stranded DNA in the presence of GSH and that treatment of formamidopyrimidine-DNA glycosylase led to chain cleavages at the guanine residues. ESR spin-trapping studies showed that 1:2:2:1 quartet DMPO (5,5-dimethyl-1-pyrroline N-oxide) spectrum similar to DMPO/hydroxy radical (*OH) adduct, but the signals were not inhibited by ethanol. Therefore, the signal seemed not to be due to *OH but byproduct due to oxidation of DMPO by the reactive species. The signals were suppressed by the addition of dGMP, but not by other mononucleotides, suggesting the specific reactivity with guanine. On the basis of our results and previous literature, it is speculated that reduction of KBrO3 by SH compounds in renal proximal tubular cells yields bromine oxides and bromine radicals, which are the reactive species that cause guanine oxidation, leading to renal carcinogenesis of KBrO3. PMID- 11409939 TI - Metabolic activation of benzo[c]phenanthrene by cytochrome P450 enzymes in human liver and lung. AB - The environmentally occurring polycyclic aromatic hydrocarbon (PAH) benzo[c]phenanthrene (B[c]PH) is a weak carcinogen in rodents. In contrast, the dihydrodiol-epoxides of B[c]PH are among the most carcinogenic PAH metabolites tested so far. In rodents, B[c]PH is predominantly metabolized to B[c]PH-5,6 dihydrodiol (B[c]PH-5,6-DH) and only to a minor extent to B[c]PH-3,4-DH, the proximate precursor of the highly potent ultimate carcinogen, B[c]PH-3,4-DH-1,2 epoxide. This might explain why in rodents B[c]PH is a weak carcinogen. However, little is known about human metabolism of B[c]PH. Using microsomal preparations from human liver and lung, we investigated the metabolic activation of B[c]PH. In contrast to the findings in experimental animals, human liver microsomes predominantly generated B[c]PH-3,4-DH and only to a minor extent B[c]PH-5,6-DH. Only one lung tissue sample was found to be metabolically active, producing B[c]PH-5,6-DH together with small amounts of B[c]PH-3,4-DH. Catalytic activities known to be associated with specific cytochrome P450 (P450) enzyme activities were determined and correlated with the spectrum of B[c]PH metabolites. The results indicate that B[c]PH-DH formation in human liver is mainly mediated by P450 1A2. Studies with P450 enzyme selective inhibitors confirmed these findings. Further support was obtained using preparations of the respective human recombinant P450 enzymes expressed in Escherichia coli and yeast. In addition to P450 1A2, P450 1B1 effectively mediated B[c]PH-metabolism. The umu-assay for induction of SOS repair response in Salmonella typhimurium TA 1535 pSK 1002 containing a umuC-lacZ reporter gene was used to study metabolic generation of genotoxic metabolites from B[c]PH-DHs in human microsomal preparations. B[c]PH 3,4-DH was activated by human liver microsomes to a potent genotoxic agent. Taken together, the results clearly demonstrate that human liver microsomes can effectively catalyze the biotransformation of B[c]PH into highly genotoxic metabolites. The results provide evidence that B[c]PH should be considered a potentially potent carcinogen in humans, and that rodent models may underestimate the risk. PMID- 11409940 TI - Indirect cytotoxicity of flucloxacillin toward human biliary epithelium via metabolite formation in hepatocytes. AB - Flucloxacillin, an isoxazolyl-penicillin, causes cholestasis and biliary epithelium injury. The aim of the study was to determine whether flucloxacillin, either directly or through metabolite formation, may induce cytotoxicity in hepatic or biliary cells. Cytotoxicity was assessed by lactate dehydrogenase release in primary cultures of human hepatocytes and of gallbladder-derived biliary epithelial cells (BEC). Metabolite production in microsome and cell preparations was analyzed by chromatography, nuclear magnetic resonance spectroscopy, and mass spectrometry. While flucloxacillin induced no direct cytotoxicity in any of the hepatocyte (n = 12) and BEC (n = 19) preparations, the conditioned media from cultured hepatocytes preincubated with flucloxacillin (50 500 mg/L) triggered a significant increase in lactate dehydrogenase release over controls in approximately 50% of BEC preparations (7/12), and this effect depended upon flucloxacillin concentration. Remaining BEC preparations exhibited no toxic response. Cytotoxicity in BEC preparations (9/13) was also induced by the supernatants of human liver microsomes and of recombinant human cytochrome P450 (CYP)3A4 preincubated with flucloxacillin (500 mg/L). Supernatants from both liver microsome and CYP3A4 preparations contained one major metabolite which was identified as 5'-hydroxymethylflucloxacillin. The production of this metabolite was inhibited following CYP3A4 inhibition by troleandomycin in human liver microsomes, and markedly enhanced following CYP3A induction by dexamethasone in rat liver microsomes. As opposed to BEC, cultured hepatocytes displayed significant CYP3A activity and produced low amounts of this metabolite. The purified metabolite (0.01-5 mg/L) exerted toxic effects in BEC but not in hepatocytes. In conclusion, hepatocytes mainly via CYP3A4 activity, generate flucloxacillin metabolite(s) including 5'-hydroxymethylflucloxacillin that may induce cytotoxicity in susceptible BEC. These metabolic events may contribute to the pathogenesis of drug-induced cholangiopathies. PMID- 11409941 TI - Analysis of urinary metabolites of tea catechins by liquid chromatography/electrospray ionization mass spectrometry. AB - Tea has been proposed to have beneficial health effects which have been attributed to the polyphenolic compounds known as catechins. The bioavailability and biotransformation of these compounds, however, are not clearly understood. In this study, we used liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS) to determine urinary glucuronidated and sulfated tea catechins and their metabolites (including methylated and ring-fission metabolites) based on the detection of deprotonated molecular ions and aglycone fragment ions. The compound resolution was achieved both chromatographically and mass spectroscopically. After green tea administration, the major conjugates appeared in human, mouse, and rat urine samples were identified as monoglucuronides and monosulfates of (-)-epigallocatechin (EGC) and (-) epicatechin. We also found O-methyl-EGC-O-glucuronides and -O-sulfates and O methyl-epicatechin-O-sulfates in human urine. (-)-5-(3',4',5'-Trihydroxyphenyl) gamma-valerolactone (M4) and (-)-5-(3',4'-dihydroxyphenyl)-gamma-valerolactone (M6), the ring-fission metabolites of EGC and (-)-epicatechin, respectively, were also predominantly in monoglucuronide and monosulfate forms in the urine. In comparison to rats, the urinary metabolite profiles of tea catechins in mice resemble more closely to those in humans. This is the first report describing direct simultaneous analysis of multiple tea catechin conjugates in urine samples. This method will allow more thorough investigations of the biotransformation of tea polyphenols. PMID- 11409942 TI - O(6)-allyl protected deoxyguanosine adducts of polycyclic aromatic hydrocarbons as building blocks for the synthesis of oligonucleotides. AB - We describe a synthetic strategy for the preparation of oligonucleotides using N(2)-alkylated and O(6)-allyl protected deoxyguanosine phosphoramidite building blocks derived from cis- and trans-opened (+/-)-7beta,8alpha-dihydroxy 9alpha,10alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene and (+/-)-7beta,8alpha dihydroxy-9beta,10beta-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene and from trans opened (+/-)-3alpha,4beta-dihydroxy-1alpha,2alpha-epoxy-1,2,3,4 tetrahydrobenzo[c]phenanthrene. The appropriately blocked phosphoramidite building blocks were obtained as mixtures of the cis- and trans-opened diol epoxide adducts upon initial reaction of the diol epoxides with O(6)-allyl-3',5' di-O-(tert-butyldimethylsilyl)-2'-deoxyguanosine. Key to the present approach is the removal of the O(6)-allyl protecting group utilizing a palladium catalyst prior to release of the constructed oligonucleotide with ammonia from the solid support. The methodology described enables a very convenient access to oligonucleotides containing cis- and trans-N(2)-deoxyguanosine adducts of polycyclic aromatic hydrocarbons in different sequence contexts. PMID- 11409943 TI - Differences between the mutational consequences of replication of cis- and trans opened benzo[a]pyrene 7,8-diol 9,10-epoxide-deoxyguanosine adducts in M13mp7L2 constructs. AB - The four adducts at N(2) of deoxyguanosine derived from cis-opening at C-10 of four optically active isomers of 7,8-dihydroxy-9,10-epoxy-7,8,9,10 tetrahydrobenzo[a]pyrene were incorporated into 5'-TTCGAATCCTTCCCCC [context III(G)] and 5'-GGGGTTCCCGAGCGGC [context IV(G)] at the underlined site. The mutagenic consequences of these lesions in each of the two sequence contexts were examined after ligation of the modified oligonucleotides into single-stranded M13mp7L2 and replication of the vector in SOS-induced Escherichia coli. Total frequencies of base substitution mutations ranged between 14 and 48%. The mutation frequencies were generally higher in context IV(G) than in context III(G), and consisted mainly of G-->T followed by G-->C base substitutions. A substantial number of deletions or insertions of one guanine was also found for all adducts in context IV(G), where the adduct is located at the 3'-end of a run of five guanines. The overall frequencies of base substitution mutations induced by cis-opened adducts were substantially higher than those observed with the trans-opened dGuo adducts in the same sequences [Page et al. (1998) Biochemistry 37, 9127-9137]. Although G-->T base substitutions predominated for both the cis- and trans-opened adducts, the cis-opened dGuo adducts generally resulted in a higher proportion of G-->C [particularly in context III(G)] relative to G-->A, whereas the opposite was true for the trans-opened dGuo adducts. The present results along with previous data indicate that mutagenicity is highly dependent on a combination of sequence context and adduct stereochemistry. PMID- 11409944 TI - Reduction of aflatoxin B1 dialdehyde by rat and human aldo-keto reductases. AB - Oxidation of the mycotoxin aflatoxin (AF) B1 yields the 8,9-epoxide, which nonenzymatically hydrolyzes rapidly to a dihydrodiol that in turn undergoes slow, base-catalyzed ring opening to a dialdehyde [Johnson, W. W., Harris, T. M., and Guengerich F. P. (1996) J. Am. Chem. Soc. 118, 8213-8220]. AFB1 dialdehyde does not bind to DNA but can react with protein lysine groups. One enzyme induced by cancer chemopreventive agents is AFB1 aldehyde reductase (AFAR), which catalyzes the NADPH-dependent reduction of the dialdehyde to a dialcohol. AFB1 dialdehyde is known to convert nonenzymatically to AFB1 dihydrodiol at neutral pH, and we reinvestigated the enzymatic reaction by preparing AFB1 dialdehyde at pH 10 and then used this to initiate reactions (at neutral pH) with rat and human AFAR isozymes. Two monoalcohols were identified as products, and their identities were established by NaB2H4 reduction, chemical cleavage, and mass spectrometry. The monoalcohol corresponding to reduction at C-8 formed first in reactions catalyzed by either the rat or the human AFAR. This C-8 monoalcohol was further reduced to AFB1 dialcohol by AFAR. The other monoalcohol (C-6a) was formed but not reduced to the dialcohol rapidly. Steady-state kinetic parameters were estimated for the reduction of AFB1 dialdehyde by rat and human AFAR to the monoalcohols. The apparent k(cat) and K(m) values were not adequate to rationalize the observed DeltaA(340) spectral changes in a kinetic model. Simulation fitting was done and yielded parameters indicative of greater enzyme efficiency. A survey of 12 human liver cytosol samples showed a variation of 2.3-fold in AFAR activity. Rats treated with AFB1 excreted the dialcohol and a monoalcohol in urine. The results of these studies are consistent with a role of (rat and human) AFAR in protection against AFB1 toxicity. PMID- 11409945 TI - Isolation and mass spectrometric characterization of dimeric adenine photoproducts in oligodeoxynucleotides. AB - UVC irradiation of oligodeoxynucleotides (ODNs) at 254 nm generates two types of DNA photoproducts, A=A and (AA)*, at adjacent adenine sites in DNA. Kumar et al. previously proved the structure of these adducts in dinucleoside monophosphates [Kumar, S., et al. (1987) Nucleic Acids Res. 15, 1199-1216; Kumar, S., et al. (1991) Nucleic Acids Res. 19, 2841-2847]. Product-ion spectra of ESI-produced [M 2H](2-) ions of the ODNs containing the dimeric adenine photoproducts show distinctive fragmentation that is informative of the structures of the photoproducts. The gas-phase cleavages of ODNs at sites of those photoproducts and thymine thymine or thymine adenine ((TA)*) photoproducts are analogous to cleavages induced by hot alkaline treatment. Nuclease-P1 digestions of ODNs containing dimeric adenine photoproducts give shorter pieces of ODNs bearing the photoproducts, which fragment under collisional activation conditions in a similar way to the large ODNs containing the photoproducts. The tandem mass spectrometric results show that the yield of (AA)* is lower than that of A=A when adjacent adenines are in the middle of an ODN sequence, and the yield of the latter is similar to that of (TA)*. PMID- 11409946 TI - Synthesis and characterization of oligodeoxynucleotides containing an N1 beta hydroxyalkyl adduct of 2'-deoxyinosine. AB - Hydroxyethyl adducts arising by the reactions of simple epoxides at the N1 position of adenine nucleosides can deaminate to give the inosine analogues which, if formed in DNA, are suspected of being highly mutagenic. A method has been developed for synthesis of oligonucleotides containing N1-adducted 2' deoxyinosines. The 2'-deoxyinosine adduct of 3,4-epoxy-1-butene was prepared from (+/-)-4-acetoxy-3-bromo-1-butene and tetraisopropyldisiloxanediyl-protected 2' deoxyinosine with base. The 2'-deoxyinosine derivative was then incorporated into the oligodeoxynucleotide sequence 5'-d(CGGACXAGAAG)-3' (X = N1-(1-hydroxy-3-buten 2-yl)-2'-deoxyinosine). PMID- 11409947 TI - Synthesis and reactivity of the catechol metabolites from the equine estrogen, 8,9-dehydroestrone. AB - The risk factors for women developing breast and endometrial cancers are all associated with a lifetime of estrogen exposure. Estrogen replacement therapy in particular has been correlated with an increased cancer risk. Previously, we showed that the equine estrogens equilin and equilenin, which are major components of the widely prescribed estrogen replacement formulation Premarin, are metabolized to highly cytotoxic quinoids which caused oxidative stress and alkylation of DNA in vitro [Bolton, J. L., Pisha, E., Zhang, F., and Qiu, S. Chem. Res. Toxicol. 1998, 11, 1113-1127]. In this study, we have synthesized 8,9 dehydroestrone (a third equine estrogen component of Premarin) and its potential catechol metabolites, 4-hydroxy-8,9-dehydroestrone and 2-hydroxy-8,9 dehydroestrone. Both 2-hydroxy-8,9-dehydroestrone and 4-hydroxy-8,9 dehydroestrone were oxidized by tyrosinase or rat liver microsomes to o-quinones which reacted with GSH to give one mono-GSH conjugate and two di-GSH conjugates. Like endogenous estrogens, 8,9-dehydroestrone was primarily converted by rat liver microsomes to the 2-hydroxylated rather than the 4-hydroxylated o-quinone GSH conjugates; the ratio of 2-hydroxy-8,9-dehydroestrone versus 4-hydroxy-8,9 dehydroestrone was 6:1. Also in contrast to experiments with equilin, 4 hydroxyequilenin was not observed in microsomal incubations with 8,9 dehydroestrone or its catechols. The behavior of 2-hydroxy-8,9-dehydroestrone was found to be more complex than 4-hydroxy-8,9-dehydroestrone as GSH conjugates resulting from 2-hydroxy-8,9-dehydroestrone were detected even without oxidative enzyme catalysis. Under physiological conditions, 2-hydroxy-8,9-dehydroestrone isomerized to 2-hydroxyequilenin to form the very stable 2-hydroxyequilenin catechol; however, 4-hydroxy-8,9-dehydroestrone was found to be stable under similar conditions. Finally, preliminary studies conducted with the human breast tumor S-30 cell lines demonstrated that the catechol metabolites of 8,9 dehydroestrone were much less toxic than 4-hydroxyequilenin (20-40-fold). These results suggest that the catechol metabolites of 8,9-dehydroestrone may have the ability to cause cytotoxicity in vivo primarily through formation of o-quinones; however, most of the adverse effects of Premarin estrogens are likely due to formation of 4-hydroxyequilenin o-quinone from equilin and equilenin. PMID- 11409952 TI - Geographical characterization of italian extra virgin olive oils using high-field (1)H NMR spectroscopy. AB - 1H high-field nuclear magnetic resonance (NMR) was used to analyze 216 extra virgin olive oils collected in three years (1996, 1997, and 1998) in different Italian areas in order to evaluate the potential contribution of this technique to the geographical characterization of olive oils. A statistical procedure performed on the intensity of selected NMR peaks has been proposed. Tree clustering analysis of NMR data performed without any a priori hypothesis showed the existence of reliable parameters able to group the olive oils according to the location of olive oil production. Linear discriminant analysis applied to selected NMR parameters of olive oils of the same year of production allowed the grouping of samples according to their geographical origin with only very few errors. Moreover, a satisfactory grouping is reached by combining the NMR data of olive oils from two different years (1996 and 1997). Operating on appropriate sampling, a careful analysis of data yielded the conclusion that the place of olive production could be singled out as a discriminating factor regardless of the cultivars from which the olive oils are derived. PMID- 11409953 TI - Development of an enzyme-linked immunosorbent assay for the determination of maduramicin in broiler chicken tissues. AB - Maduramicin is one of the most widely used coccidiostats in the world. A rapid and accurate analytical method for this drug should provide producers and users with an effective management tool. The current chromatographic methods are sensitive but labor-intensive. This paper reports the development of an enzyme linked immunosorbent assay (ELISA) based on an immunoaffinity chromatography cleanup procedure for the analysis of maduramicin in broiler chicken tissues (including muscle, liver, and fat). Recoveries from fortified tissue homogenates at levels of 30.0-120.0 microg kg(-)(1) ranged from 76.4 to 107.5% with coefficients of variation of 3.8-16.4%. The limits of detection were 1.0 ng g( )(1) in muscle, 2.8 ng g(-)(1) in liver, and 1.5 ng g(-)(1) in fat. The ELISA results from the analysis of incurred residue in tissue samples showed the cleanup procedure is viable. PMID- 11409954 TI - Development of NMR and raman spectroscopic methods for the determination of the degree of substitution of maleate in modified starches. AB - In this paper we report the application of NMR spectroscopy and Raman spectroscopy to determine the degree of maleate substitution in maleinated starches. Five kinds of maleinated starches were investigated and calibration sets were constructed to derive linear regression equations that may be used to predict the degree of maleate substitution for starch samples with unknown amounts of chemical modification. The calibration sets reported have very high linearity (r > 0.99) for both the NMR and Raman methods. The NMR and Raman calibration sets allow fast and nondestructive measurement of the degree of maleate substitution for different starches with little need of sample preparation. PMID- 11409955 TI - Degradation of lignin in wheat straw during growth of the oyster mushroom (Pleurotus ostreatus) using off-line thermochemolysis with tetramethylammonium hydroxide and solid-state (13)C NMR. AB - The oyster mushroom (Pleurotus ostreatus) is widely cultivated on wheat straw (Triticum aestivum); however, there is a need to better understand the relationship between the chemical composition of the compost and mushroom growth. Wheat straw was degraded over a period of 63 days by P. ostreatus during which time it was sampled at weekly intervals. Off-line thermochemolysis with tetramethylammonium hydroxide and solid-state (13)C NMR were then used in the molecular characterization of the undegraded wheat straw and the degraded samples. The degraded wheat straw samples had a lower proportion of syringyl- to guaiacyl-derived moieties and cinnamyl- to guaiacyl-derived moieties than the undegraded control. There were increases in both guaiacyl and syringyl acid to aldehyde ratios with composting time, which showed that side-chain oxidation has been mediated by P. ostreatus. The (13)C NMR spectra confirmed the increase in carboxyl content but indicated that the overall lignin and methoxyl contents remained relatively constant, although some nonsystematic variations were observed. The spectra also showed a decrease in amorphous noncellulosic polysaccharides in relation to the crystalline cellulose upon degradation. PMID- 11409956 TI - Identification of goose (Anser anser) and mule duck (Anasplatyrhynchos x Cairina moschata) foie gras by multiplex polymerase chain reaction amplification of the 5S RDNA gene. AB - Polymerase chain reaction (PCR) amplification of the nuclear 5S rDNA gene has been used for the identification of goose and mule duck foie gras. Two species specific reverse primers were designed and used in a multiplex reaction, together with a forward universal primer, to amplify specific fragments of the 5S rDNA in each species. The different sizes of the species-specific amplicons, separated by agarose gel electrophoresis, allowed clear identification of goose and mule duck foie gras samples. This genetic marker can be useful for detecting fraudulent substitution of the duck liver for the more expensive goose liver. PMID- 11409957 TI - Determination of organic acids, sugars, diacetyl, and acetoin in cheese by high performance liquid chromatography. AB - Sugars (lactose, glucose, and galactose), nonvolatile acids (citric, orotic, piruvic, lactic, ossalic, and hippuric), some free fatty acids (formic, acetic, propionic, butyric, isobutyric, valeric, and isovaleric), diacetyl, and acetoin were separated on an Aminex HPX-87H column using a simple isocratic HPLC method and identified by retention times with ultraviolet and refractive index detectors. With the proposed technique it is possible to evaluate the development of microbial fermentations and, at the same time, degree of cheese ripening. PMID- 11409958 TI - Kinetics method for the quantitation of anthocyanidins, flavonols, and flavones in foods. AB - Flavonoids are important dietary constituents owing to their health-promoting properties. As a result, simplified analytic techniques are required for the population of databases with food values so that associations between dietary intake and disease risk/incidence can be established. The current research provides a simplified sample preparation procedure for the accurate estimation of food anthocyanidins, flavones, and flavonols as aglycons. Traditionally, flavonoid aglycons have been formed by acidic hydrolysis. However, some flavonoid aglycons are slowly degraded by acid. A procedure has been developed whereby anthocyanidins and flavonols are deglycosylated with HCl in 50% aqueous methanol and the resulting aglycons subsequently quantified by application of pseudo-first order kinetics to their degradation. Flavones are also deglycosolated under similar conditions but, at appropriate temperatures, their aglycons are stable in acid, so kinetics were not required for the quantitation of this subclass of flavonoids. Catechins and flavanones were rapidly degraded under the hydrolytic conditions used in these studies. PMID- 11409959 TI - Assay of ochratoxin A in wine and beer by high-pressure liquid chromatography photodiode array and gas chromatography mass selective detection. AB - To routinely assay the concentrations of ochratoxin A (OTA) in wines and beers, two new methods were developed and evaluated. The first utilized solid-phase extraction on a C(18) cartridge to achieve a 100-fold sample concentration followed by high-performance liquid chromatography on a C(18) column with gradient elution and quantitation at 333 nm by means of a photodiode array detector. Positive confirmation can be carried out by purity and match-factor analysis as well as peak shift following esterification with BF(3). Total run time is 28 min. The limits of detection (LOD) and quantitation (LOQ) are 0.05 and 0.10 microg/L, respectively. Recovery and imprecision ranged from 83 to 94% and from 4.0 to 8.9%, respectively. With a throughput of 35 assays per working day, this method is ideal for routine OTA analysis. It was used to survey the concentrations of OTA in 942 wines (2 of which gave values between 0.1 and 0.2 microg/L) and 107 beers (2 of which gave values between 0.05 and 0.1 microg/L). OTA was detected more frequently in red than white wines, with the highest incidence in red wines from Spain and Argentina. There was no association between OTA and country of origin or beverage type among the beers analyzed. The second method utilized gas chromatography with mass selective detection monitoring eight specific ions, preceded by extraction in dichloromethane and derivatization with bis[trimethylsilyl]trifluoroacetamide. LOD and LOQ were 0.1 and 2 microg/L, respectively; recovery and imprecision were 69-75 and 9.0-11.1%, respectively. The method is not suitable for routine quantitation but is potentially useful as a confirmatory tool for samples with OTA > or =0.1 microg/L. PMID- 11409960 TI - Solid phase extraction gas chromatography/electron capture detector method for the determination of organochlorine pesticides in wildlife whole blood. AB - A gas chromatographic method for the analysis of 10 organochlorine pesticides in 0.5 mL of whole blood is described. Sample preparation involved an ethyl ether and hexane extraction, followed by a silica solid phase extraction cleanup. The pesticides are quantified by gas chromatography/electron capture detection. Method limits of detection ranged from 1.1 to 5.2 microg/L. The mean and standard deviation for the recovery of 10 pesticides was 97.9 +/- 5.5%. Recoveries from whole blood were comparable to recoveries from plasma. This indicates that the preparation of plasma is unnecessary for the quantification of organochlorine pesticides in blood. This approach is particularly useful as a nonlethal approach for monitoring pesticide contamination in small animals for which the volume of blood is limiting. PMID- 11409961 TI - Electrospray liquid chromatography quadrupole ion trap mass spectrometry determination of phenyl urea herbicides in water. AB - Phenyl urea herbicides were determined in water by electrospray quadrupole ion trap liquid chromatography-mass spectrometry (ES-QIT-LC-MS). Over a wide concentration range [M - H](-) and MH(+) ions were prominent in ES spectra. At high concentrations dimer and trimer ions appeared, and sodium, potassium, and ammonium adducts also were observed. In the case of isopturon, source collision induced dissociation (CID) fragmentation with low offset voltages increased the ion current associated with MH(+) and diminished dimer and trimer ion abundance. In the mass analyzer CID involved common pathways, for example, daughter ions of [M - H](-) resulted from loss of R(2)NH in N',N'-dialkyl ureas or loss of C(3)H(5)NO(2) (87 amu) in N'-methoxy ureas. A 2 mm (i.d.) x 15 cm C(18) reversed phase column was used for LC-MS with a linear methanol/water gradient and 0.5 mL/min flow rate. Between 1 and 100 pg/microg/L the response was highly linear with instrument detection limits ranging from <10 to 50 pg injected. Whereas the positive ES signal intensity was greater for each of the compounds except fluometuron, negative ion monitoring gave the highest signal-to-noise ratio. Analysis of spiked Colorado River water, a source high in total dissolved solids and total organic carbon, demonstrated that ES-QIT-LC-MS was routinely capable of quantitative analysis at low nanogram per liter concentrations in conjunction with a published C(18) SPE method. Under these conditions experimental method detection limits were between 8.0 and 36 ng/L, and accuracy for measurements in the 20-50 parts per trillion range was from 77 to 96%. Recoveries were slightly lower in surface water (e.g., 39-76%), possibly due to suppression of ionization. PMID- 11409962 TI - Improved method for determination of pectin degree of esterification by diffuse reflectance Fourier transform infrared spectroscopy. AB - An improved method for the determination of pectin degree of esterification (DE) by diffuse reflectance Fourier transform infrared spectroscopy (DRIFTS) was developed. Pectin samples with a range of DE as determined by gas chromatography were used for developing a calibration curve by DRIFTS. A linear relationship between the DE of pectin standards and FTIR peak ratio for ester carboxyl peak area to total carboxyl peak area was found (R(2) = 0.97). Pectin DE of various samples was calculated from the linear fit equation developed by DRIFTS. Accuracy of the DRIFTS method was determined by comparing the DE values of four commercial pectins obtained by DRIFTS methods to the values obtained by the gas chromatography method. Greater precision was obtained for the FTIR measurement of test pectin samples when the ester peak ratio was used relative to the ester peak area. PMID- 11409963 TI - Supercritical CO(2) and subcritical propane extraction of pungent paprika and quantification of carotenoids, tocopherols, and capsaicinoids. AB - Ground paprika (Capsicum annuum L.) was extracted with supercritical carbon dioxide (SC-CO(2)) and subcritical propane at different conditions of pressure and temperature to estimate the yield and variation in carotenoid, tocopherol, and capsaicinoid contents and composition. The yield of paprika extract was found to be affected by the extraction conditions with SC-CO(2) but fairly constant at different conditions with subcritical propane. The maximum yields of oleoresin were 7.9 and 8.1% of ground paprika by SC-CO(2) and subcritical propane, respectively. The quantitative distribution of carotenoids, tocopherols, and capsaicinoids between paprika extract and powder was influenced by extraction conditions. SC-CO(2) was inefficient in the extraction of diesters of xanthophylls even at 400 bar and 55 degrees C, whereas tocopherols and capsaicinoids were easy to extract at these conditions. Under mild conditions subcritical propane was superior to SC-CO(2) in the extraction of carotenoids and tocopherols but less efficient in the extraction of capsaicinoids. PMID- 11409964 TI - Isolation and characterization of structurally novel antimutagenic flavonoids from spinach (Spinacia oleracea). AB - Thirteen compounds, isolated from spinach (Spinacia oleracea), acted as antimutagens against the dietary carcinogen 2-amino-3-methylimidazo[4,5 f]quinoline in Salmonella typhimurium TA 98. The antimutagens were purified by preparative and micropreparative HPLC from a methanol/water (70:30, v/v) extract of dry spinach (commercial product) after removal of lipophilic compounds such as chlorophylls and carotenoids by solid-phase extraction (SPE). Pure active compounds were identified by instrumental analysis including FT-IR, (1)H and (13)C NMR, UV-vis spectroscopy, and mass spectrometry. All of these compounds were flavonoids and related compounds that could be attributed to five groups: (A, methylenedioxyflavonol glucuronides) 5,3'-dihydroxy-4'-methoxy-6,7 methylenedioxyflavonol 3-O-beta-glucuronide (compound 1), 5,2',3'-trihydroxy-4' methoxy-6,7-methylenedioxyflavonol 3-O-beta-glucuronide (compound 2), 5-hydroxy 3',4'-dimethoxy-6,7-methylenedioxyflavonol 3-O-beta-glucuronide (compound 3); (B, flavonol glucuronides) 5,6,3'-trihydroxy-7,4'-dimethoxyflavonol 3-O-beta glucuronide (compound 4), 5,6-dihydroxy-7,3',4'-trimethoxyflavonol 3-O-beta glucuronide (compound 5); (C, flavonol disaccharides) 5,6,4'-trihydroxy-7,3' dimethoxyflavonol 3-O-disaccharide (compound 6), 5,6,3',4'-tetrahydroxy-7 methoxyflavonol 3-O-disaccharide (compounds 7 and 8); (D, flavanones) 5,8,4' trihydroxyflavanone (compound 9), 7,8,4'-trihydroxyflavanone (compound 10); (E, flavonoid-related compounds) compounds 11, 12, and 13 with incompletely elucidated structures. The yield of compound 1 was 0.3%, related to dry weight, whereas the yields of compounds 2-13 ranged between 0.017 and 0.069%. IC(50) values (antimutagenic potencies) of the flavonol glucuronides ranged between 24.2 and 58.2 microM, whereas the flavonol disaccharides (compounds 7 and 8), the flavanones (compounds 9 and 10), and the flavonoid-related glycosidic compounds 11-13 were only weakly active. The aglycons of compounds 7 and 8, however, were potent antimutagens (IC(50) = 10.4 and 13.0 microM, respectively). PMID- 11409965 TI - Antioxidant and antiradical activities of flavonoids. AB - The relationship between the structure of 42 flavonoids and their antioxidant and antiradical activities was elucidated by heat-induced oxidation in a beta carotene and linoleic acid system and by the 1,1-diphenyl-2-picrylhydrazyl decoloration test. From seven structurally divergent groups of flavonoids, only flavonols with a free hydroxyl group at the C-3 position of the flavonoid skeleton showed high inhibitory activity to beta-carotene oxidation. Antiradical activity depended on the presence of a flavonol structure or free hydroxyl group at the C-4' position. The effect of the 4'-hydroxyl was strongly modified by other structural features, such as the presence of free hydroxyls at C-3 and/or C 3' and a C2-C3 double bond. PMID- 11409966 TI - Conjugated triene oxidation products of alpha-farnesene induce symptoms of superficial scald on stored apples. AB - The sesquiterpene alpha-farnesene (1) and its conjugated triene oxidation products accumulate in the skin of apples after harvest and are implicated as the causal agents of superficial scald. Conjugated triene oxidation products and analogues were synthesized and applied to the surface of Granny Smith apples either as vapors or in squalane. Farnesyl hydroperoxide (2a), trienol (2b), endoperoxide (3a), dehydronerolidol (5), and cumyl hydroperoxide (4) all produced the symptoms of superficial scald when applied at nanomolar doses. Scald-inducing activity was dependent on the mode of application. Farnesyl hydroperoxide (2a) was the most active conjugated triene when applied directly to the apple skin. Trienol (2b) also induced scald symptoms and partially reversed the inhibition of scald caused by diphenylamine. PMID- 11409967 TI - Isolation and characterization of novel benzoates, cinnamates, flavonoids, and lignans from Riesling wine and screening for antioxidant activity. AB - A German Riesling wine has been fractionated with the aid of countercurrent chromatography. After purification by HPLC, the structures of 101 compounds were established by mass spectrometry and NMR spectroscopy. Seventy-three of the isolated compounds exhibited a phenolic or benzylic structure. Fifty-four compounds were reported for the first time as Riesling wine constituents. New compounds identified in this work included twelve benzoic and cinnamic acid derivatives. In addition to two isomeric (E)-caffeoyl ethyl tartrates, the glucose esters of (E)-cinnamic, (E)-p-coumaric, and (E)-ferulic acid, as well as the 4-O-glucosides of (E)- and (Z)-ferulic acid, have been identified for the first time in Riesling wine. The structures of two additional phenylpropanoids were elucidated as 3-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-propan-1-one and 2,3 dihydroxy-1-(4-hydroxy-3-methoxyphenyl)-propan-1-one. Moreover, two ethyl esters, i.e., ethyl protocatechuate and ethyl gallate, as well as the glucose ester of vanillic acid, were newly detected in Riesling wine. Novel representatives in the flavonoid group were dihydrokaempferol, dihydroquercetin, and four dihydroflavonol glycoconjugates, i.e., the 3-O-glucosides of dihydrokaempferol and dihydroquercetin, as well as the 3-O-xyloside and the 3'-O-glucoside of dihydroquercetin. Additionally, six novel lignans, i.e., lariciresinol 4-O glucoside, three isolariciresinol derivatives, and two secoisolariciresinols, as well as three neolignans were isolated. Structural elucidation of the newly isolated wine constituents is reported together with the determination of their antioxidant activity. PMID- 11409968 TI - Broad-spectrum antimicrobial activity in vitro of the synthetic peptide D4E1. AB - Broad-spectrum antimicrobial activity of a synthetic peptide, D4E1, is documented in this paper. D4E1 inhibited the growth of several fungal phytopathogens belonging to four classes-Ascomycetes, Basidiomycetes, Deuteromycetes, and Oomycetes, and two bacterial pathogens, Pseudomonas syringae pv. tabaci and Xanthomonas campestris pv. malvacearum race 18. The minimum inhibitory concentration (MIC) of D4E1 required to completely inhibit the growth of all fungi studied ranged from 4.67 to 25 microM. Fungal pathogens highly sensitive to D4E1 include Thielaviopsis basicola, Verticillium dahliae, Fusarium moniliforme, Phytophthora cinnamomi, and Phytophthora parasitica. Comparatively, the least sensitive fungal pathogens were Alternaria alternata, Colletotrichum destructivum, and Rhizoctonia solani. The two bacterial pathogens, P. syringae pv. tabaci and X. campestris pv. malvacearum race 18, were most sensitive to D4E1 with MIC values of 2.25 and 1.25 microM, respectively. Microscopic analysis of D4E1 effects on fungal morphology of Aspergillus flavus and R. solani revealed abnormal hyphal growth and discontinuous cytoplasm. After 8 h of exposure to 25 microM D4E1, A. flavus spore germination was reduced by 75%. The suitability of peptide D4E1 to enhance disease resistance in transgenic crop plants is discussed. PMID- 11409969 TI - Effects of pretreatments on the diffusion kinetics and some quality parameters of osmotically dehydrated apple slices. AB - This study compared mass transfer during osmotic dehydration (OD) and some quality indices of untreated apple slices to those of apple slices pretreated by either blanching, freezing, or applying high-intensity electric field pulses (HELP) or high pressure (HP). HP, HELP, and blanching increased water loss. Untreated and HELP-treated samples had comparable solids gains, which were lower (P < 0.05) than in the other samples. Apple slices turned brown after pretreatment but the L values of these samples increased with OD. The breaking force of dried samples increased with OD time, and pretreated samples had firmer dried texture than the untreated. Vitamin C content decreased with OD time, but HP- and HELP-treated apples had better retention of vitamin C. PMID- 11409970 TI - Bipolar membrane electroacidification of demineralized skim milk. AB - The aim of this study was to evaluate the effect of decreasing the mineral content of skim milk by electrodialysis (ED) prior to electroacidification with bipolar membrane (BMEA) on the performance of the process, the chemical composition, and the physicochemical and functional properties of the isolates produced. ED used to demineralize the skim milk solution was very efficient. However, the electroacidification parameters were influenced by the demineralization level of the skim milk solution: the energy efficiency was decreased with an increase in demineralization, but it was still possible to perform BMEA at a very low conductivity level. Moreover, the isolates produced by BMEA after electrodialysis demineralization at different rates showed similar chemical composition, except on potassium and lactose contents for 75% demineralized isolate. These isolates, except on protein load for 75% demineralization rate, showed similar physicochemical and functional properties, whatever the demineralization rate. PMID- 11409971 TI - Copper(II) catalysis in cyanide conversion into ethyl carbamate in spirits and relevant reactions. AB - The role of copper(II) species in the oxidation of inorganic cyanide to cyanate and in the conversion of cyanate or urea into ethyl carbamate was investigated. The oxidation process has been shown to be independent from the dissolved oxygen. Elemental analysis and infrared spectroscopy have shown the formation of a mixed copper carbonate/hydroxide in the process of oxidation of cyanide to cyanate in water/ethanol. The complexation to Cu(II) of cyanate formed upon cyanide oxidation makes the former more susceptible to nucleophilic attack from ethanol, with conversion into ethyl carbamate. Comparatively, urea has a minor role with respect to cyanide in the formation of ethyl carbamate. Therefore, the urea present in some samples of Brazilian sugar cane spirit (cachaca) has been shown to have almost no influence on the ethyl carbamate content of cachacas, which comes essentially from cyanide. Fe(II,III) affords results similar to those found with Cu(II). Some suggestions are presented to avoid ethyl carbamate formation in spirits during distillation. PMID- 11409972 TI - Revising the role of pH and thermal treatments in aflatoxin content reduction during the tortilla and deep frying processes. AB - Naturally aflatoxin-contaminated corn (Zea mays L.) was made into tortillas, tortilla chips, and corn chips by the traditional and commercial alkaline cooking processes. The traditional nixtamalization (alkaline-cooking) process involved cooking and steeping the corn, whereas the commercial nixtamalization process only steeps the corn in a hot alkaline solution (initially boiling). A pilot plant that includes the cooker, stone grinder, celorio cutter, and oven was used for the experiments. The traditional process eliminated 51.7, 84.5, and 78.8% of the aflatoxins content in tortilla, tortilla chips, and corn chips, respectively. The commercial process was less effective: it removed 29.5, 71.2, and 71.2 of the aflatoxin in the same products. Intermediate and final products did not reach a high enough pH to allow permanent aflatoxin reduction during thermal processing. The cooking or steeping liquor (nejayote) is the only component of the system with a sufficiently high pH (10.2-10.7) to allow modification and detoxification of aflatoxins present in the corn grain. The importance of removal of tip, pericarp, and germ during nixtamalization for aflatoxin reduction in tortilla is evident. PMID- 11409973 TI - Reaction of B. cereus bacteria and peroxidase enzymes under pressures >400 MPa. AB - It is known that B. cereus (Gram-positive bacteria) and peroxidase enzymes are resistant to pressures of approximately 400 MPa in fruit and vegetable products among others. The aim of the present work is to have knowledge about their behavior when using pressures >400 MPa without other combined treatments. The results showed that B. cereus was not inactivated at pressures of 1000 MPa for 15 min at 20 degrees C. In peroxidase enzymes the results remained similar and the pressure of 1000 MPa for 30 min was not enough to inactivate them. The apple cell structure at these high-pressure levels revealed that it changed and the cells were less cemented. The treated apple presented a translucent aspect, and some fluids migrated from the inside to the outside of the cell. PMID- 11409974 TI - Sulfur compounds reduce potato toxins during extrusion cooking. AB - Free sulfhydryl groups in sulfur compounds have been reported to act directly on natural toxins to reduce toxicity. The objective of this study was to reduce protease inhibitors and glycoalkaloids in simulated snack foods by the addition of sulfur-containing compounds prior to extrusion. Thiamine, methionine, and benzyl disulfide were added to potato flakes at levels of 0.5% or 1.0% prior to twin-screw extrusion. Total and free thiols and protease inhibitors were monitored before and after extrusion by colorimetric assays. Potato glycoalkaloids were analyzed by HPLC and by immunoassay. Extrusion reduced potato flake disulfide bonds; disulfide bonds were higher in samples containing added sulfur compounds. Trypsin inhibitor activity was reduced by as much as 79% by extrusion plus methionine. Extrusion significantly reduced carboxypeptidase inhibitor, but only when benzyl disulfide and 0.5% methionine were not added. One percent methionine and thiamine resulted in 60% reductions in glycoalkaloids. PMID- 11409975 TI - Changes in the profile of genistein, daidzein, and their conjugates during thermal processing of tofu. AB - Profiles of genistein, daidzein, genistin, daidzin, and their acetyl- and malonyl beta-glycosides were determined in tofu as affected by temperature and time. Tofu was heated in water at 80, 90, and 100 degrees C for 0 (control), 10, 20, 30, and 40 min, and the contents of the isoflavones of interest were quantified using reversed-phase HPLC. Total isoflavone content decreased most likely due to leaching of isoflavones into the water. Because the content of the isoflavones of the genistein series was little affected by the treatments, the decrease in the total isoflavone content was almost exclusively due to a decrease of the daidzein series. Changes in the profile of the daidzein series suggest little decarboxylation of the malonylglycoside to the acetylglycoside, but considerable de-esterification of the malonyl- and acetylglycoside to the beta-glucoside. Strongly temperature dependent decreases of the aglycon suggest possible thermal degradation of daidzein in addition to losses due to leaching. PMID- 11409976 TI - Stability of chlorpyrifos for termiticidal control in six Australian soils. AB - Chlorpyrifos [O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate] is the most widely used soil-applied termiticide in Australia. It is relatively stable, has low water solubility, is absorbed by organic matter, and has a high affinity for soil with low partitioning potential from soil matter to soil water. The purpose of this degradation study is to determine the effect of soil alkalinity on the longevity of termite protection when chlorpyrifos is applied as a termiticide in a range of Australian soils, particularly high-pH substrates. The study also examines the effects of initial soil concentration on the degradation of chlorpyrifos in the range of soils. At an initial soil concentration of 1000 mg kg(-)(1) for termite control, the degradation rate of chlorpyrifos is very strongly retarded in soils tested when compared with lower soil concentrations of 100 and 10 mg kg(-)(1) in the same soils. The degradation data correlated with a logarithmic model of decay, and it was thus possible to produce half-lives and predict likely periods of termite control. Average half-lives for all soils for the three concentrations were 385, 155, and 41 days, respectively. Soil pH had no effect on the rate of degradation at all concentrations tested. PMID- 11409977 TI - Ion chromatographic determination of the decomposition products of tecnazene solution irradiated by ultraviolet light: inorganic and organic anions. AB - Ion chromatographic (IC) methods using sodium hydroxide and methanol gradients were used for the determination of small inorganic and organic anions as decomposition products of a tecnazene solution in water irradiated by UV light. After 60 min of UV irradiation, >99% of the tecnazene was decomposed, and 11 organic and 3 inorganic anions were identified and quantified. A fourth inorganic anion, carbonate, was not quantified due to likely losses as carbon dioxide. The final content of chloride, total nitrogen, and total carbon released from the saturated tecnazene solution after 60 min of UV irradiation were 108, 85, and 38%, respectively. These results suggest that other products rich in carbon and/or nitrogen (such as phenolic compounds and nitrobenzenes) were formed during the UV irradiation of the tecnazene solution. The results obtained indicate several decomposition pathways of tecnazene in water solutions, for example, ring opening reactions, dechlorination, and replacement of the ring nitro group. The determination of nonionic decomposition products from the latter two possibilities is the subject of further study. PMID- 11409978 TI - Uniformly (14)C-ring-labeled 2,4-dichlorophenoxyacetic acid: a metabolism study in bluegill sunfish, Lepomis macrochirus. AB - The fate of 2,4-dichlorophenoxyacetic acid (2,4-D), a mixture of [phenyl(U) (14)C]-2,4-D and unlabeled 2,4-D, in bluegill sunfish was investigated after exposure to approximately 11 ppm under static conditions for 4 days. Total radioactive residues (TRR) in whole fish increased from 0.41 ppm on day 1 to 0.60 ppm on day 3. TRR levels in fillet (edible) and viscera (nonedible) of treated fish on day 4 were 0.41 and 1.9 ppm, respectively. Most residues in both matrices were acetonitrile soluble; small amounts were hexane soluble or unextractable with solvents. Acid and base hydrolyses with ethyl acetate partitioning were used to release the fillet unextractable residues. The identification of 2,4-D and 2,4 dichlorophenol (2,4-DCP) in the fillet was conclusively confirmed by GC-MS analysis. On the basis of the experimental data from this study, a metabolic pathway for 2,4-D in bluegill sunfish in which the 2,4-D is metabolized to 2,4 DCP and conjugates of 2,4-D and 2,4-DCP is proposed. PMID- 11409979 TI - Effects of lamination and coating with drying oils on tensile and barrier properties of zein films. AB - Zein films plasticized with oleic acid have been considered potentially useful for biodegradable packaging applications. However, moisture was found to affect their tensile and gas barrier properties. We investigated the effects of two converting processes, fusion lamination and coating with drying oils, on tensile properties and gas permeability of zein films. Zein films were laminated to 4-ply sheets in a Carver press and coated with tung oil, linseed oil, or a mixture of tung and soybean oils. Tensile properties and permeability to water vapor, oxygen, and carbon dioxide were measured according to ASTM methods. Laminated films were clearer, tougher, and more flexible, and had a smoother finish than nontreated sheets. Lamination decreased O(2) and CO(2) permeability by filling in voids and pinholes in the film structure. Coating increased tensile strength and elongation and decreased water vapor permeability. Coatings acted as a composite layer preventing crack propagation and increasing film strength. They also formed a highly hydrophobic surface that prevented film wetting. PMID- 11409980 TI - Relative abundance and inhibitory distribution of protease inhibitors in potato juice from cv. Elkana. AB - Protease inhibitors from potato juice of cv. Elkana were purified and quantified. The protease inhibitors represent ca. 50% of the total soluble proteins in potato juice. The protease inhibitors were classified into seven different families: potato inhibitor I (PI-1), potato inhibitor II (PI-2), potato cysteine protease inhibitor (PCPI), potato aspartate protease inhibitor (PAPI), potato Kunitz-type protease inhibitor (PKPI), potato carboxypeptidase inhibitor (PCI), and "other serine protease inhibitors". The most abundant families were the PI-2 and PCPI families, representing 22 and 12% of all proteins in potato juice, respectively. Potato protease inhibitors show a broad spectrum of enzyme inhibition. All the families (except PCI) inhibited trypsin and/or chymotrypsin. PI-2 isoforms exhibit 82 and 50% of the total trypsin and chymotrypsin inhibiting activity, respectively. A strong variation within the latter activities was shown within one family and between protease inhibitor families. PMID- 11409981 TI - Improved method for the analysis of organic acids and new derivatization of alcohols in complex natural aqueous matrixes: application to wine and apple vinegar. AB - An improvement in the procedure for investigation of organic acids and a new derivatization method, amenable to gas chromatographic-mass spectrometric detection of alcohols, are presented. The latter is based on the formation of phenacetyl esters. The simultaneous application of the two methods also allows data to be obtained on some volatile neutrals present in complex natural aqueous fluids. Application to wine and cider vinegars allowed detection of a number of previously unreported components, among which are interesting partially esterified polycarboxylic esters. PMID- 11409982 TI - Metabolism of phenolic compounds during loquat fruit development. AB - Phenolic compounds in loquat fruit were identified as 5-caffeoylquinic acid (chlorogenic acid), neochlorogenic acid, hydroxybenzoic acid, 5-p-feruloylquinic acid, protocatechuic acid, 4-caffeoylquinic acid, epicatechin, o-coumaric acid, ferulic acid, and p-coumaric acid. Neochlorogenic acid was found to be dominant in the early stages of loquat fruit development. Both the concentrations and types of phenolic compounds were high in young fruit but then decreased steadily during growth. However, the concentration of chlorogenic acid increased during ripening and became predominant in ripe fruit. The large rise in chlorogenic acid concentration appears to be a characteristic of loquat fruit ripening. In all of the cultivars tested, the types of phenolic compounds were similar but the total phenolic content varied from 81.8 to 173.8 mg/100 g of fresh pulp. In the biosynthetic pathway of chlorogenic acid, the enzyme activities of phenylalanine ammonia-lyase (PAL), 4-coumarate:CoA ligase (CL), and hydroxycinnamoyl CoA:quinate hydroxycinnamoyl transferase (CQT) were high at the early stage of growth, diminished to low levels approximately 3 weeks prior to harvest, but then rose to a peak at 1 week before harvest. The changes of these enzyme activities seemed to be associated with variations in chlorogenic acid concentration during development, maturation, and ripening of loquat fruit. PMID- 11409983 TI - Modeling of the sorption and diffusion processes of volatile organic air pollutants in grape fruits. AB - A mathematical model for the description of the sorption and diffusion processes of gaseous toluene and p-xylene in fruits of grape has been proposed. This model is based on the Fick's II low regarding a particle with a spherical shape (such as the berry of grapevine) and describes changes of air pollutant concentrations in different layers of the fruit, that is, the wax, peel, and pulp, during an exposure to contaminated air. The mass transfer coefficient and diffusion coefficients in the respective layers can be estimated using the experimental values. The theoretical data and the results from the exposure under steady-state laboratory conditions were compared and showed a good applicability of the proposed model for the prediction of volatile air pollutant partitioning in grape berries. PMID- 11409984 TI - Mobility and dissipation of ethofumesate and halofenozide in turfgrass and bare soil. AB - The effect of turfgrass cover on the leaching and dissipation of ethofumesate and halofenozide was studied. Sampling cylinders (20 cm diam. x 30 cm long) were placed vertically in plots of creeping bentgrass (Agrostis palustris Huds.), tall fescue (Festuca arundinaceae Schreb.), or bare soil. ethofumesate [(+/-)-2-ethoxy 2,3-dihydro-3,3-dimethylbenzofuran-5-yl methansulfonate] was applied at 840 g ai ha(-)(1) on September 21, 1997. Halofenozide (N-4-chlorobenzoyl-N'-benzoyl-N' tert-butylhydrazine) was applied at 1680 g ai ha(-)(1) on August 30, 1998. Replicate sampling cylinders were removed 2 h after treatment and 4, 8, 16, 32, and 64 days after treatment. Sampling cylinders were sectioned by depths and soil extracts were assayed by HPLC with a pesticide detection limit of 0.01 mg kg( )(1). Turfgrass was divided into verdure and thatch and analyzed separately. ethofumesate leaching in turfgrass was reduced by at least 95% compared to leaching in bare soil. The half-life of ethofumesate in bare soil was 51 days compared to 3 days in turfgrass. Halofenozide showed similar leaching with or without turfgrass. Fifty percent dissipation of halofenozide did not occur within 64 days, regardless of organic matter cover. PMID- 11409985 TI - Potential contributions of smectite clays and organic matter to pesticide retention in soils. AB - Soil organic matter (SOM) is often considered the dominant sorptive phase for organic contaminants and pesticides in soil-water systems. This is evidenced by the widespread use of organic-matter-normalized sorption coefficients (K(OM)) to predict soil-water distribution of pesticides, an approach that ignores the potential contribution of soil minerals to sorption. To gain additional perspective on the potential contributions of clays and SOM to pesticide retention in soils, we measured sorption of seven pesticides by a K-saturated reference smectite clay (SWy-2) and SOM (represented by a muck soil). In addition, we measured the adsorption of atrazine by five different K-saturated smectites and Ca-saturated SWy-2. On a unit mass basis, the K-SWy-2 clay was a more effective sorbent than SOM for 4,6-dinitro-o-cresol (DNOC), dichlobenil, and carbaryl of the seven pesticides evaluated, of which, DNOC was sorbed to the greatest extent. Atrazine was sorbed to a similar extent by K-SWy-2 and SOM. Parathion, diuron, and biphenyl were sorbed to a greater extent by SOM than by K SWy-2. Atrazine was adsorbed by Ca-SWy-2 to a much lesser extent than by K-SWy-2. This appears to be related to the larger hydration sphere of Ca(2+) (compared to that of K(+)) which shrinks the effective size of the adsorption domains between exchangeable cations, and which expands the clay layers beyond the apparently optimal spacing of approximately 12.2 A for sorption of aromatic pesticide structures. Although a simple relation between atrazine adsorption by different K smectites and charge properties of clay was not observed, the highest charge clay was the least effective sorbent; a higher charge density would result in a loss of adsorption domains. These results indicate that for certain pesticides, expandable soil clays have the potential to be an equal or dominant sorptive phase when compared to SOM for pesticide retention in soil. PMID- 11409986 TI - Organ- and season-dependent variation in the essential oil composition of Salvia officinalis L. cultivated at two different sites. AB - More than 50 compounds were identified in essential oils from stems and leaves of Salvia officinalis L. plants harvested in July, in Arouca, in northern Portugal. About 40 of those compounds were also present in flower essential oils, collected from the same plants. alpha-Thujone was the major compound, representing about 55, 30, and 18% of the essential oils from stems, leaves, and flowers, respectively. Significant percentage variations in the main compound classes of the essential oils from shoots sampled over the year were recorded at two different sites in northern Portugal. From December to April, oxygenated monoterpenes (MO) decreased from approximately 67-72% to values of 42-43% of the essential oils. During the same time interval, the percentage of monoterpene hydrocarbons (MH) rose from 8-11% to 17-22%. At both sites, sesquiterpene hydrocarbons (SH) rose from approximately 7% in February to 19-22% in April, decreasing thereafter to approximately 9% in July. Oxygenated sesquiterpenes (SO) increased from a minimum of approximately 5% in July to a maximum of 8-11% in February, decreasing thereafter. The compounds that mostly accounted for the essential oil composition variation were alpha-pinene, beta-pinene, and camphene, as MH; alpha-thujone and camphor, as MO; alpha-humulene and beta-caryophyllene, as SH; and viridiflorol, as SO. PMID- 11409987 TI - Volatile compounds released by enzymatic hydrolysis of glycoconjugates of leaves and grape berries from Vitis vinifera Muscat of Alexandria and Shiraz cultivars. AB - Glycoconjugates from Muscat of Alexandria and Shiraz leaves and grape berries were isolated by adsorption on Amberlite XAD-2 resin, and enzymatically released aglycons were analyzed by GC-FID and GC-MS. About 120 aglycons were fully or tentatively identified. Compositional differences were observed between leaves and berries of the two varieties in five aglycon chemical groups: C6 alcohols, aliphatic alcohols, monoterpenes, shikimates, and C(13)-norisoprenoids, which were much more abundant in the leaves than in the berries. The differences observed for C(13)-norisoprenoids were in agreement with their hypothetical independent biosynthesis in leaves and berries. Thus, 3-hydroxy-beta-damascone, an important norisoprenoid aglycon of grape berries, was not detected in leaves, whereas its oxidized derivative, 3-oxo-alpha-damascone, was absent in berries. Compositional differences were also observed between Muscat and Shiraz leaves. 3 Oxo-alpha-ionol was not detected in Shiraz leaves, and its retro derivatives were less abundant than in Muscat of Alexandria leaves. Conversely, in Shiraz leaves the levels of 7,8-dihydroionone derivatives, such as megastigman-3,9-diol and 3 oxo-7,8-dihydro-alpha-ionol, were higher than in Muscat of Alexandria leaves. PMID- 11409988 TI - Identification and quantification of impact odorants of aged red wines from Rioja. GC-olfactometry, quantitative GC-MS, and odor evaluation of HPLC fractions. AB - An XAD-4 extract from a 5-year-old wine from Rioja (Spain) was analyzed by aroma extract dilution analysis. Most of the odorants were quantified by GC-MS. A second extract was fractionated in an HPLC system with a C-18 semipreparative column. Fifty fractions were recovered, their alcoholic degree and pH were further adjusted to those of the wine, and those fractions that showed strong odor characteristics were further re-extracted and analyzed by GC-O and GC-MS. Reconstitution experiments were carried out to confirm the role of the odorants detected in the fractions. Fifty-eight odorants were found in the Rioja wine, 52 of which could be identified. Methyl benzoate was found to be a wine aroma constituent for the first time. The most important odorants are 4-ethylguaiacol, (E)-whiskey lactone, 4-ethylphenol, beta-damascenone, fusel alcohols, isovaleric and hexanoic acids, eugenol, fatty acid ethyl esters, and ethyl esters of isoacids, Furaneol, phenylacetic acid, and (E)-2-hexenal. Comparison among the three techniques shows good agreement and demonstrates that they are complementary. PMID- 11409989 TI - Evolution of the taste of a bitter Camembert cheese during ripening: characterization of a matrix effect. AB - The objective of this study was to characterize the effect of ripening on the taste of a typically bitter Camembert cheese. The first step was to select a typically bitter cheese among several products obtained by different processes supposed to enhance this taste defect. Second, the evolution of cheese taste during ripening was characterized from a sensory point of view. Finally, the relative impact of fat, proteins, and water-soluble molecules on cheese taste was determined by using omission tests performed on a reconstituted cheese. These omission tests showed that cheese taste resulted mainly from the gustatory properties of water-soluble molecules but was modulated by a matrix effect due to fat, proteins, and cheese structure. The evolution of this matrix effect during ripening was discussed for each taste characteristic. PMID- 11409990 TI - Evolution of the composition of a selected bitter Camembert cheese during ripening: release and migration of taste-active compounds. AB - The aim of this study was to add to the understanding of changes in taste that occur during the ripening of a bitter Camembert cheese by the evolution of its composition. Physicochemical analyses were performed on rind, under-rind, and center portions of a Camembert cheese selected for its intense bitterness. At each of the six steps of ripening studied organic acids, sugars, total nitrogen, soluble nitrogen, phosphotungstic acid soluble nitrogen, non-protein nitrogen, Na, K, Ca, Mg, Pi, Cl, and biogenic amines were quantified in each portion. Changes in cheese composition seemed to mainly result from the development of Penicillium camemberti on the cheese outer layer. Migration phenomena and the release of potentially taste-active compounds allowed for the evolution of saltiness, sourness, and bitterness throughout ripening to be better understood. Apart from taste-active compounds, the impact of the cheese matrix on its taste development is discussed. PMID- 11409991 TI - Aroma-active components of nonfat dry milk. AB - Application of aroma extract dilution analysis (AEDA) on the volatile components of low-, medium-, and high-heat-treated nonfat dry milks (NDM) revealed aroma active compounds in the log(3) flavor dilution (log(3) FD) factor range of 1 to 6. The following compounds contributed the highest log(3) FD factors to overall NDM flavor: 2,5-dimethyl-4-hydroxy-3(2H)-furanone [(Furaneol), burnt sugar-like]; butanoic acid (rancid); 3-(methylthio)propanal [(methional), boiled potato-like]; o-aminoacetophenone (grape-like); delta-decalactone (sweet); (E)-4,5-epoxy-(E)-2 decenal (metallic); pentanoic acid (sweaty); 4,5-dimethyl-3-hydroxy-2(5H) furanone [(sotolon), curry]; 3-methoxy-4-hydroxybenzaldehyde [(vanillin), vanilla]; 2-acetyl-1-pyrroline and 2-acetyl-2-thiazoline (popcorn-like); hexanoic acid (vinegar-like); phenylacetic acid (rose-like); octanoic acid (waxy); nonanal (fatty); and 1-octen-3-one (mushroom-like). The odor intensities of Furaneol, butanoic acid, methional, o-aminoacetophenone, sotolon, vanillin, (E)-4,5-epoxy (E)-2-decenal, and phenylacetic acid were higher in high-heat-treated samples than others. However, the odor intensities of lactones, 2-acetyl-1-pyrroline, and 2-acetyl-2-thiazoline were not affected by heat treatment. Sensory evaluation results also revealed that heat-generated flavors have a major impact on the flavor profile of NDM. PMID- 11409992 TI - Enzymatic synthesis of vanillin. AB - Due to increasing interest in natural vanillin, two enzymatic routes for the synthesis of vanillin were developed. The flavoprotein vanillyl alcohol oxidase (VAO) acts on a wide range of phenolic compounds and converts both creosol and vanillylamine to vanillin with high yield. The VAO-mediated conversion of creosol proceeds via a two-step process in which the initially formed vanillyl alcohol is further oxidized to vanillin. Catalysis is limited by the formation of an abortive complex between enzyme-bound flavin and creosol. Moreover, in the second step of the process, the conversion of vanillyl alcohol is inhibited by the competitive binding of creosol. The VAO-catalyzed conversion of vanillylamine proceeds efficiently at alkaline pH values. Vanillylamine is initially converted to a vanillylimine intermediate product, which is hydrolyzed nonenzymatically to vanillin. This route to vanillin has biotechnological potential as the widely available principle of red pepper, capsaicin, can be hydrolyzed enzymatically to vanillylamine. PMID- 11409993 TI - Identification of potent odorants formed by autoxidation of arachidonic acid: structure elucidation and synthesis of (E,Z,Z)-2,4,7-tridecatrienal. AB - The aroma composition of autoxidized arachidonic acid was characterized by aroma extract dilution analysis. The most potent odorant was trans-4,5-epoxy-(E)-2 decenal followed by 1-octen-3-one, (E,Z)-2,4-decadienal, (E,Z,Z)-2,4,7 tridecatrienal, (E,E)-2,4-decadienal, and hexanal. (E,Z,Z)-2,4,7-Tridecatrienal was unequivocally identified by mass spectrometry and nuclear magnetic resonance (NMR) data. The stereochemistry of its extended double-bond system was elucidated on the basis of NMR measurements. The target compound was synthesized in four steps starting with bromination of 2-octyn-1-ol, followed by copper-catalyzed coupling of the bromide with ethylmagnesium bromide and (E)-2-penten-4-yn-1-ol. Partial hydrogenation of the resulting C(13)-compound with triple bonds in the positions C-4 and C-7 gave rise to (E,Z,Z)-2,4,7-tridecatrien-1-ol, which was finally oxidized to the target compound. It exhibits a typical egg-white-like, marine-like odor at low concentrations, and an intense orange-citrus, animal-like odor at higher concentrations. Its odor threshold was estimated by gas chromatography-olfactometry to be 0.07 ng/L air, which is of the same order of magnitude as that reported for 1-octen-3-one and (E,E)-2,4-decadienal. PMID- 11409994 TI - Biomass nutrient profiles of the microalga Nannochloropsis. AB - The nutritional composition of the marine eustigmatophyte Nannochloropsis spp. cultured in an indoor chemostat under continuous illumination was analyzed. Proximate composition, (moisture, ash, crude protein, available carbohydrates, fiber, lipids, and energy), nitrate, nucleic acid, mineral element (Na, K, Ca, Mg, Fe, Cu, Zn, Mn, Pb, Cd, Cr, Ni, Co, and S), fatty acid, and pigment (carotenoids and chlorophyll) concentrations were determined. On average, the biomass contained 37.6% (w/w) available carbohydrates, 28.8% crude protein, and 18.4% total lipids. Mineral in 100 g of dry biomass were as follows: Ca (972 mg), K (533 mg), Na (659 mg), Mg (316 mg), Zn (103 mg), Fe (136 mg), Mn (3.4 mg), Cu (35.0 mg), Ni (0.22 mg), and Co (<0.1 mg). Toxic heavy metal contents (Cd and Pb) were negligible. Fatty acid content was as follows (on percent dry weight): 0.6% of 14:0, 5.0% of 16:0; 4.7% of 16:1omega7, 3.8% of 18:1omega9, 0.4% of 18:2omega6; 0.7% of 20:4omega6, and 2.2% of 20:5omega3. Nutrient composition of the biomass was highly influenced by residence time in the photobioreactor. The biomass harvested for short residence times was richer in protein and eicosapentaenoic acid than biomass harvested for high residence time. PMID- 11409995 TI - Experimental approach to optimize the use of alpha-Amylases in breadmaking. AB - alpha-Amylases from different origins (wheat, malted barley, fungi, and bacteria) are used extensively to improve breadmaking. However, the enzyme activities, in addition to the differences associated with their origins, are strongly affected by the process conditions and the presence of other compounds in the medium. The activity of different alpha-amylases was tested under different conditions (pH and temperature), and in the presence of some bread ingredients (salt and sugar), some breadmaking additives (ascorbic acid and sodium propionate), and some metabolites (organic acids and saccharides) generated during the fermentation step, to envisage the behavior of these alpha-amylases during the breadmaking process. The alpha-amylase activities were affected to a different extent by the addition of these compounds depending on the enzyme origin. In general, the alpha amylases from cereals (wheat and malted barley) were less sensitive to the presence of some ingredients, additives, and metabolites. These results show the great variation of the alpha-amylase activity with the process conditions and the importance of its knowledge in the selection of the appropriate alpha-amylase for a specific breadmaking process. PMID- 11409996 TI - Antioxidant activities of trypsin inhibitor, a 33 KDa root storage protein of sweet potato (Ipomoea batatas (L.) Lam cv. Tainong 57). AB - Trypsin inhibitors (TIs), root storage proteins, were purified from sweet potato (Ipomoea batatas[L.] Lam cv. Tainong 57) roots by trypsin affinity column according to the methods of Hou and Lin (Plant Sci. 1997, 126, 11-19 and Plant Sci. 1997, 128, 151-158). A single band of 33 kDa TI was obtained by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels. This purified 33 kDa TI had scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. There was positive correlation between scavenging effects against DPPH (2 to 22%) and amounts of 33 kDa TI (1.92 to 46 pmol). The scavenging activities of 33 kDa TI against DPPH were calculated from linear regression to be about one-third of those of glutathione between 5 and 80 pmol. Using electron paramagnetic resonance (EPR) spectrometry for hydroxyl radical detection, it was found that 33 kDa TI could capture hydroxyl radical, and the intensities of EPR signal were significantly decreased from 1.5 to 6 pmol of 33 kDa TI compared to those of the controls. It is suggested that 33 kDa TI, one of the sweet potato root storage proteins, may play a role as an antioxidant in roots and may be beneficial to health when it is consumed. PMID- 11409997 TI - m-Iodobenzoic acid complexes with selected metals: molecular structure and antimicrobial activity. AB - Complexes of lithium, sodium, potassium, rubidium, cesium, magnesium, calcium, manganese, and zinc with m-iodobenzoic acid were studied. The FT-IR and FT-Raman spectra of the mentioned compounds in the solid state and water solutions were recorded and analyzed. Principal component analysis (PCA) was performed on the wavenumbers of selected bands (eight bands) occurring in the vibrational spectra. The numbers obtained as a result of this procedure characterize the electronic properties of the molecule of each complex. The antimicrobial activity of the studied compounds against selected bacteria (Escherichia coli and Bacillus subtilis) and yeast (Saccharomyces cerevisiae and Hansenula anomala) was estimated. The relationship between the chemical properties (as characterized by PCA of the IR spectra) and antimicrobial properties of the compounds was examined, and a good correlation between the two factors was found. PMID- 11409998 TI - Conformational changes and some functional characteristics of gelatin esterified with fatty acid. AB - This study investigated the effects of attachment of fatty acid chains to gelatin molecules on their conformation and some functional properties in order to determine the effectiveness of this procedure for improving the emulsifying properties of gelatin. The esterification conferred upon the gelatin molecules a folded configuration resulting in a lambda(max) of fluorescence emission shift to lower wavelengths and decreases in solubility, intrinsic viscosity, and gelling ability. In contrast, surface hydrophobicity increased with extent of esterification, and the esterified derivatives exhibited higher emulsion stabilities. These results indicated that the oligomeric structure and functional attributes of the gelatin were altered with the modification. PMID- 11409999 TI - Angiotensin I converting enzyme inhibitory peptides purified from bovine skin gelatin hydrolysate. AB - Bovine skin gelatin was hydrolyzed with sequential protease treatments in the order of Alcalase, Pronase E, and collagenase using a three-step ultrafiltration membrane reactor. The molecular weight distributions of the first, second, and third hydrolysates were 4.8-6.6, 3.4-6.6, and 0.9-1.9 kDa, respectively. The angiotensin I converting enzyme (ACE) inhibitory activity of the third hydrolysate (IC(50) = 0.689 mg/mL) was higher than that of the first and second hydrolysates. Two different peptides showing strong ACE inhibitory activity were isolated from the hydrolysate using consecutive chromatographic methods including gel filtration chromatography, ion-exchange chromatography, and reversed-phase high-performance liquid chromatography. The isolated peptides were composed of Gly-Pro-Leu and Gly-Pro-Val and showed IC(50) values of 2.55 and 4.67 microM, respectively. PMID- 11410000 TI - Oxidation of ascorbate in raw milk induced by enzymes and transition metals. AB - The effect of xanthine oxidase, lactoperoxidase, and transition metals [Fe(III), Cu(II)] on the oxidation of ascorbate in raw milk was investigated. Data clearly showed that iron(III) (200 microM) does not accelerate ascorbate oxidation in raw milk in concentrations relevant for raw milk. In contrast, addition of copper(II) (10 microM) to the raw milk accelerated oxidation of ascorbate. Furthermore, both xanthine oxidase and peroxidase activity were found to accelerate ascorbate oxidation dramatically in raw milk, indicating that xanthine oxidase and lactoperoxidase might be some of the most obvious candidates for mediation of ascorbate oxidation in raw milk. The present data are discussed in relation to using the fate of ascorbate in raw milk as an indicator of the oxidative stability of the milk. PMID- 11410001 TI - His-His-Leu, an angiotensin I converting enzyme inhibitory peptide derived from Korean soybean paste, exerts antihypertensive activity in vivo. AB - It has been reported that soybean peptide fractions isolated from Korean fermented soybean paste exert angiotensin I converting enzyme (ACE) inhibitory activity in vitro. In this study, further purification and identification of the most active fraction inhibiting ACE activity were performed, and its antihypertensive activity in vivo was confirmed. Subsequently, a novel ACE inhibitory peptide was isolated by preparative HPLC. The amino acid sequence of the isolated peptide was identified as His-His-Leu (HHL) by Edman degradation. The IC(50) value of the HHL for ACE activity was 2.2 microg/mL in vitro. Moreover, the synthetic tripeptide HHL (spHHL) resulted in a significant decrease of ACE activity in the aorta and led to lowered systolic blood pressure (SBP) in spontaneously hypertensive (SH) rats compared to control. Triple injections of spHHL, 5 mg/kg of body weight/injection resulted in a significant decrease of SBP by 61 mmHg (p < 0.01) after the third injection. These results demonstrated that the ACE inhibitory peptide HHL derived from Korean fermented soybean paste exerted antihypertensive activity in vivo. PMID- 11410002 TI - Effects of temperature and agitation rate on the formation of conjugated linoleic acids in soybean oil during hydrogenation process. AB - The effects of hydrogen temperature and agitation rate on the formation of total conjugated linoleic acids (CLA) and CLA isomers were studied during hydrogenation with a selective Ni catalyst. The CLA isomers were identified by using a 100-m cyano-capillary column gas chromatograph and a silver ion-impregnated HPLC. Reaction temperature and agitation rate greatly affected the quantities of total CLA and individual CLA isomers, and the time to reach the maximum quantity of CLA in the partially hydrogenated soybean oil. As the hydrogenation temperature increased, the maximum quantity of CLA in soybean oil increased, but the time to reach the maximum CLA content decreased. By increasing the hydrogenation temperature from 170 to 210 degrees C, the quantity of CLA obtained was about 2.6 times higher. As the agitation rate decreased, the CLA formation in soybean oil increased, and the time to reach the maximum CLA content also increased. The maximum CLA contents in soybean oil obtained during hydrogenation at 210 degrees C with agitation rates of 300, 500, and 700 rpm were 162.82, 108.62, and 66.15 mg total CLA/g oil, respectively. The present data showed that it is possible to produce high-CLA-content soybean oil without major modification of fatty acid composition by short-time (10 min) selective hydrogenation under high temperature and low agitation rate conditions. PMID- 11410003 TI - Elisa to quantify hexanal-protein adducts in a meat model system. AB - Monoclonal antibodies (MAb) were produced to hexanal-bovine serum albumin conjugates. An indirect competitive ELISA was developed with a detection range of 1-50 ng of hexanal/mL. Hexanal conjugated to three different proteins was recognized, whereas free hexanal and the native proteins were not detected. The antibody cross-reacted with pentanal, heptanal, and 2-trans-hexenal conjugated to chicken serum albumin (CSA) with cross-reactivities of 37.9, 76.6, and 45.0%, respectively. There was no cross-reactivity with propanal, butanal, octanal, and nonanal conjugated to CSA. The hexanal content of a meat model system was determined using MAb and polyclonal antibody-based ELISAs and compared with analysis by a dynamic headspace gas chromatographic (HS-GC) method and a thiobarbituric acid reactive substances (TBARS) assay. Both ELISAs showed strong correlations with the HS-GC and TBARS methods. ELISAs may be a fast and simple alternative to GC for monitoring lipid oxidation in meat. PMID- 11410004 TI - Oxidative metabolism of the soy isoflavones daidzein and genistein in humans in vitro and in vivo. AB - The soy isoflavones daidzein and genistein are found in high concentrations in human plasma and urine after soy consumption. However, in vitro and in vivo data regarding the oxidative metabolism of isoflavones in humans are scarce. Therefore, we have studied the oxidative metabolites of these compounds formed in human liver microsomes and excreted in urine of male and female humans ingesting soy products for 2 days. Human liver microsomes transformed the soy isoflavone daidzein to three monohydroxylated and three dihydroxylated metabolites according to GC/MS analysis. On the basis of a previous study with rat liver microsomes and with the help of reference substances, these metabolites were identified as 6,7,4'-trihydroxyisoflavone, 7,3',4'-trihydroxyisoflavone, 7,8,4' trihydroxyisoflavone, 7,8,3',4'-tetrahydroxyisoflavone, 6,7,8,4' tetrahydroxyisoflavone, and 6,7,3',4'-tetrahydroxyisoflavone. Significant amounts of the same metabolites except 6,7,8,4'-tetrahydroxyisoflavone were also found in urine of female and male volunteers after soy intake. Genistein was metabolized by human liver microsomes to six hydroxylation products. The main metabolites were the three aromatic monohydroxylated products 5,6,7,4' tetrahydroxyisoflavone, 5,7,8,4'-tetrahydroxyisoflavone and 5,7,3',4' tetrahydroxyisoflavone. The aliphatic monohydroxylated metabolite 2,5,7,4' tetrahydroxyisoflavone and two aromatic dihydroxylated metabolites, 5,7,8,3',4' pentahydroxyisoflavone and 5,6,7,3',4'-pentahydroxyisoflavone, were formed in trace amounts. The same hydroxylated genistein metabolites except the aliphatic hydroxylated one could also be detected in human urine samples. Methylated forms of the catechol metabolites, which were generated by incubations with catechol-O methyltransferase in vitro could be detected only in trace amounts in the urine samples. This implies that this reaction does not play a major role in the biotransformation of the hydroxylated daidzein and genistein metabolites in vivo. Most of these oxidative metabolites are described as human in vivo metabolites for the first time. Their biological significance remains to be established. PMID- 11410005 TI - Antioxidant mechanism of flavonoids. Solvent effect on rate constant for chain breaking reaction of quercetin and epicatechin in autoxidation of methyl linoleate. AB - The rate of oxygen depletion, as measured by electron spin resonance spectroscopy (oximetry using a spin probe), in a homogeneous solution of peroxidating methyl linoleate (initiated by an azo initiator) in the presence or absence of antioxidants was converted to second-order rate constants for the inhibiting reaction of quercetin and epicatechin. In the non-hydrogen-bonding solvent chlorobenzene at 50 degrees C, k(inh) had values of 4.3 x 10(5) M(-)(1) s(-)(1) for quercetin and 4.2 x 10(5) M(-)(1) s(-)(1) for epicatechin, respectively. In the hydrogen-accepting "water-like" solvent tert-butyl alcohol, the values were 2.1 x 10(4) and 1.7 x 10(4) M(-)(1) s(-)(1), respectively. The solvent effect (factor of 20) is more significant than for alpha-tocopherol (factor of 4), and the two flavonoids have efficiencies comparable to that of alpha-tocopherol in scavenging peroxyl radicals in the nonpolar solvent but not in the hydrogen bonding solvent. PMID- 11410006 TI - Permeation of oxygen, water vapor, and limonene through printed and unprinted biaxially oriented polypropylene films. AB - Oriented polypropylene (OPP) and coated OPP (acrylic/OPP/PVDC) films were printed with two commercially available inks to investigate the influence of inks on water vapor and oxygen transmission rates. The permeation of an aroma compound (d limonene) through coated OPP film printed with these inks was also evaluated at 35 degrees C and 100% relative humidity. The water vapor transmission rate increased significantly through OPP film printed with nitrocellulose-based ink. The oxygen transmission rate was significantly lower through both OPP and coated OPP films printed with the nitrocellulose ink. The effect of inks on limonene permeation was minor compared to the marked increase in permeation measured when the PVDC side of the coated film was exposed to the aroma, compared to the acrylic side. Scanning electron micrographs of coated film cross sections revealed changes in film structure upon exposure to limonene vapors, which were most pronounced when the PVDC side was exposed to limonene. PMID- 11410007 TI - Structural analysis of a novel antimutagenic compound, 4-Hydroxypanduratin A, and the antimutagenic activity of flavonoids in a Thai spice, fingerroot (Boesenbergia pandurata Schult.) against mutagenic heterocyclic amines. AB - Six compounds were isolated from fresh rhizomes of fingerroot (Boesenbergia pandurata Schult.) as strong antimutagens toward 3-amino-1,4-dimethyl-5H pyrido[4,3-b]indole (Trp-P-1) in Salmonella typhimurium TA98. These compounds were 2',4',6'-trihydroxychalcone (pinocembrin chalcone; 1), 2',4'-dihydroxy-6' methoxychalcone (cardamonin; 2), 5,7-dihydroxyflavanone (pinocembrin; 3), 5 hydroxy-7-methoxyflavanone (pinostrobin; 4), (2,4,6-trihydroxyphenyl)-[3'-methyl 2'-(3' '-methylbut-2' '-enyl)-6'-phenylcyclohex-3'-enyl]methanone (5), and (2,6 dihydroxy-4-methoxyphenyl)-[3'-methyl-2'-(3' '-methylbut-2' '-enyl)-6' phenylcyclohex-3'-enyl]methanone (panduratin A; 6). Compound 5 was a novel compound (tentatively termed 4-hydroxypanduratin A), and 1 was not previously reported in this plant, whereas 2-4 and 6 were known compounds. The antimutagenic IC(50) values of compounds 1-6 were 5.2 +/- 0.4, 5.9 +/- 0.7, 6.9 +/- 0.8, 5.3 +/ 1.0, 12.7 +/- 0.7, and 12.1 +/- 0.8 microM in the preincubation mixture, respectively. They also similarly inhibited the mutagenicity of 3-amino-1-methyl 5H-pyrido[4,3-b]indole (Trp-P-2) and 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine (PhIP). All of them strongly inhibited the N-hydroxylation of Trp-P-2. Thus, the antimutagenic effect of compounds 1-6 was mainly due to the inhibition of the first step of enzymatic activation of heterocyclic amines. PMID- 11410008 TI - Citrus pectin: characterization and inhibitory effect on fibroblast growth factor receptor interaction. AB - This study was undertaken to characterize the pectin from four citrus species and to determine their in vitro inhibitory activities on the binding of fibroblast growth factor (FGF) to the FGF receptor (FGFR). Pectin from various parts of lemon, grapefruit, tangerine, and orange were isolated and characterized. Tangerine had the highest pectin content among the four citrus species. Segment membrane contained as much as or more pectin than flavedo/albedo. Anhydrogalacturonic content was highest in pectin from segment membrane of tangerine and flavedo/albedo of grapefruit. Lemon pectin contained the highest methoxyl content (MC), and grapefruit contained the largest proportion of lower molecular weight (<10000 Da) pectin. Tangerine contained the highest neutral sugar in both flavedo/albedo and segment membrane. The interdependency of heparin on factor-receptor interaction provides a means for identifying new antagonists of growth factor activity and thus for treatment of various diseases. These results showed that pectin significantly inhibited the binding of FGF-1 to FGFR1 in the presence of 0.1 microg/mL heparin. The pectin from the segment membrane of lemon was the most potent inhibitor. The inhibition activity was significantly correlated with sugar content, MC, and size of pectin. Kinetic studies revealed a competitive nature of pectin inhibition with the heparin, a crucial component of the FGF signal transduction process. The observation that the heparin-dependent biological activity of FGF signal transduction is antagonized by citrus pectin should be further investigated for the use of these pectins as anti-growth factor agents for potential health benefits. PMID- 11410009 TI - Effect of various domestic processing methods on antinutrients and in vitro protein and starch digestibility of two indigenous varieties of Indian tribal pulse, Mucuna pruriens Var. utilis. AB - The effect of various domestic processing methods on antinutrients and starch fractions and in vitro protein and starch digestibilities of white and black varieties of Mucuna pruriens var. utilis was studied. Cooking or autoclaving of both raw seeds and presoaked seeds in different solutions (water, tamarind extract, sodium bicarbonate, and citric acid) significantly (p < 0.05) reduced the content of total phenolics, phytic acid, trypsin inhibitor and chymotrypsin inhibitor activities, and L-dopa compared to soaking or dry heating techniques. The germination processes (24 and 48 h) were also effective in the reduction of various antinutrients, although this reduction appeared to be more pronounced in a prolonged period of germination (72 h). Water soaking followed by dehusking was found to be ineffective in the reduction of trypsin and chymotrypsin inhibitor activities in both varieties. All of the treatments were effective in significantly (p < 0.05) reducing the resistant starch content in the presently investigated samples. Cooking as well as autoclaving brought about a more significant (p < 0.05) improvement in the digestibility of protein and starch compared to germination and dry heat treatment. Moreover, among the different processing techniques, soaking in sodium bicarbonate solution followed by cooking (29.6-34.8%) or autoclaving (33.0-37.2%) seemed to be the best method for improving starch digestibility. PMID- 11410010 TI - Preparation of bean curds from protein fractions of six legumes. AB - Chickpeas, lentils, smooth peas, mung beans, and faba beans were milled into flours and fractionated to protein and starch fractions. Compositions of the seeds, cotyledons, and flours were compared for each legume and the weight and protein recovery of each fraction analyzed. Bean curds were prepared from the protein fractions through heat denaturation of protein milk, followed by coagulation with calcium sulfate or magnesium sulfate. The effect of chickpea protein concentration and coagulant dosage on the texture of bean curds was evaluated using a texture analyzer. Textural analysis indicated that curd prepared at 2.3-3.0% protein concentration and 1.5% CaSO(4) dosage had better yield and better texture than curds prepared under other conditions. Bean curds prepared from chickpeas and faba beans exhibited the second highest springiness and cohesiveness after those from soybeans. Curds of mung beans and smooth peas, on the other hand, had the highest yields and the highest moisture contents. The protein yield of the first and second soluble extracts used for curd preparation accounted for approximately 90% of the total protein of the seeds. PMID- 11410011 TI - Peroxynitrite-induced oxidation of lipids: implications for muscle foods. AB - Peroxynitrite (ONOO(-)), formed from the nearly diffusion limited reaction between nitric oxide and superoxide, could be an important prooxidant in muscle foods. The objective of this study was to determine whether peroxynitrite caused oxidation of pyrogallol red, liposomes, muscle microsomes, and skeletal muscle homogenate. Oxidation of pyrogallol red, liposomes, and microsomes initiated by peroxynitrite continuously produced by 3-morpholinosydnonimine (SIN-1, 2 mM) was time-dependent and enhanced by CO(2) (1 mM). Reagent peroxynitrite (2 mM) caused concentration-dependent oxidation of pyrogallol red, liposomes, and muscle microsomes that was very rapid with no change after 5 min. Peroxynitrite-induced oxidation was suppressed by CO(2) and low pH. Skeletal muscle homogenate oxidized by reagent peroxynitrite (0.5 mM) exhibited gradual oxidation with time and was suppressed by CO(2), low pH, and metal chelators. These data suggest that peroxynitrite could be an important prooxidant in muscle foods. PMID- 11410012 TI - Incompatibility of mixing of proteins in adsorbed binary protein films at the air water interface. AB - Competitive adsorption of proteins from several binary protein solutions to the air-water interface has been studied. With a few exceptions, the equilibrium composition of the saturated monolayer of mixed protein films at various bulk concentration ratios did not follow a Langmuir-type competitive adsorption model. The deviation from ideal behavior results from incompatibility of mixing of proteins in the film at the air-water interface. This immiscibility alters the ratio of the binding affinity of the proteins in a protein 1/protein 2/water ternary film compared to that in a protein 1/water and protein 2/water binary film. A method to determine the extent of incompatibility between two proteins in a mixed protein film has been developed. It is shown that the incompatibility index derived for 19 protein 1/water and protein 2/water systems studied show a linear relationship with the absolute difference between Flory-Huggins protein solvent interaction parameters, that is, /chi(1s) - chi(2)/, of the constituent proteins. On the basis of the evidence, it is theorized that, because of incompatibility, proteins in a mixed protein film at interfaces may undergo two dimensional phase separation. PMID- 11410013 TI - Lateral phase separation in adsorbed binary protein films at the air-water interface. AB - Lateral phase separation in two-dimensional mixed films of soy 11S/beta-casein, acidic subunits of soy 11 (AS11S)/beta-casein, and alpha-lactalbumin/beta-casein adsorbed at the air-water interface has been studied using an epifluorescence microscopy method. No distinct lateral phase separation was observed in the mixed protein films when they were examined after 24 h of adsorption from the bulk phase. However, when the soy 11S/beta-casein and AS11S/beta-casein films were aged at the air-water interface for 96 h, phase-separated regions of the constituent proteins were evident, indicating that the phase separation process was kinetically limited by a viscosity barrier against lateral diffusion. In these films, beta-casein always formed the continuous phase and the other globular protein the dispersed phase. The morphology of the dispersed patches was affected by the protein composition in the film. In contrast with soy 11S/beta casein and AS11S/beta-casein films, no lateral phase separation was observed in the alpha-lactalbumin/beta-casein film at both low and high concentration ratios in the film. The results of these studies proved that proteins in adsorbed binary films exhibit limited miscibility, and the deviation of competitive adsorption behavior of proteins at the air-water interface from that predicted by the ideal Langmuir model (Razumovsky, L.; Damodaran, S. J. Agric. Food Chem. 2001, 49, 3080 3086) is in fact due to thermodynamic incompatibility of mixing of the proteins in the binary film. It is hypothesized that phase separation in adsorbed mixed protein films at the air-water and possibly oil-water interfaces of foams and emulsions might be a source of instability in these dispersed systems. PMID- 11410014 TI - Origin of multiexponential T(2) relaxation in muscle myowater. AB - To obtain a further understanding of the nature of the multiexponential T(2) relaxation seen in muscle tissue water (myowater), relaxation measurements were carried out on whole, minced, and homogenized pork of three different qualities with regard to water-holding capacity (normal, red soft exudative, and dark firm dry). Whole, minced, and homogenized pork all resulted in multiexponential T(2) relaxation (three components) independently of the quality, even though microscopic studies on homogenized meat revealed considerable disruption of the macroscopic structure. This states that the relaxation behavior in meat cannot be explained by intra-/extracellular compartmentalization of the water as suggested in earlier studies. Subsequent studies of T(2) relaxation in either whole meat, where the structure integrity was changed by the introduction of dimethyl sulfoxide (membrane disruption) or urea (protein denaturation), or minced meat with added NaCl (inter-/intraprotein interactions) lead to the suggestion that in whole meat (i) the fastest relaxation component reflects water tightly associated with macromolecules, (ii) the intermediate relaxation component reflects water located within highly organized protein structures, for example, water in tertiary and/or quaternary protein structures and spatials with high myofibrillar protein densities including actin and myosin filament structures, and (iii) the slowest relaxation component reflects the extra-myofibrillar water containing the sarcoplasmatic protein fraction. Finally, relaxation patterns in heat-set gels of superprecipitated actomyosin and bovine serum albumin similar to that identified in whole meat support the proposed nature of T(2) relaxation in muscle myowater. PMID- 11410015 TI - Alpha-tocopherol content in 62 edible tropical plants. AB - Vitamin E was determined by the high-performance liquid chromatography (HPLC) method. All the plants tested showed differences in their alpha-tocopherol content and the differences were significant (p < 0.05). The highest alpha tocopherol content was in Sauropus androgynus leaves (426.8 mg/kg edible portion), followed by Citrus hystrix leaves (398.3 mg/kg), Calamus scipronum (193.8 mg/kg), starfruit leaves Averrhoa belimbi (168.3 mg/kg), red pepper Capsicum annum (155.4 mg/kg), local celery Apium graveolens (136.4 mg/kg), sweet potato shoots Ipomoea batatas (130.1 mg/kg), Pandanus odorus (131.5 mg/kg), Oenanthe javanica (146.8 mg/kg), black tea Camelia chinensis (183.3 mg/kg),papaya Carica papaya shoots (111.3 mg/kg), wolfberry leaves Lycium chinense (94.4 mg/kg), bird chili Capsicum frutescens leaves (95.4 mg/kg), drumstick Moringa oleifera leaves (90.0 mg/kg), green chili Capsicum annum (87 mg/kg), Allium fistulosum leaves (74.6 mg/kg), and bell pepper Capsicum annum (71.0 mg/kg). alpha-Tocopherol was not detected in Brassica oleracea, Phaeomeria speciosa, Pachyrrhizus speciosa, Pleurotus sajor-caju, and Solanum melongena. PMID- 11410016 TI - Flavonoid (myricetin, quercetin, kaempferol, luteolin, and apigenin) content of edible tropical plants. AB - Studies were conducted on the flavonoids (myricetin, quercetin, kaempferol, luteolin, and apigenin) contents of 62 edible tropical plants. The highest total flavonoids content was in onion leaves (1497.5 mg/kg quercetin, 391.0 mg/kg luteolin, and 832.0 mg/kg kaempferol), followed by Semambu leaves (2041.0 mg/kg), bird chili (1663.0 mg/kg), black tea (1491.0 mg/kg), papaya shoots (1264.0 mg/kg), and guava (1128.5 mg/kg). The major flavonoid in these plant extracts is quercetin, followed by myricetin and kaempferol. Luteolin could be detected only in broccoli (74.5 mg/kg dry weight), green chili (33.0 mg/kg), bird chili (1035.0 mg/kg), onion leaves (391.0 mg/kg), belimbi fruit (202.0 mg/kg), belimbi leaves (464.5 mg/kg), French bean (11.0 mg/kg), carrot (37.5 mg/kg), white radish (9.0 mg/kg), local celery (80.5 mg/kg), limau purut leaves (30.5 mg/kg), and dried asam gelugur (107.5 mg/kg). Apigenin was found only in Chinese cabbage (187.0 mg/kg), bell pepper (272.0 mg/kg), garlic (217.0 mg/kg), belimbi fruit (458.0 mg/kg), French peas (176.0 mg/kg), snake gourd (42.4 mg/kg), guava (579.0 mg/kg), wolfberry leaves (547.0 mg/kg), local celery (338.5 mg/kg), daun turi (39.5 mg/kg), and kadok (34.5 mg/kg). In vegetables, quercetin glycosides predominate, but glycosides of kaempferol, luteolin, and apigenin are also present. Fruits contain almost exclusively quercetin glycosides, whereas kaempferol and myricetin glycosides are found only in trace quantities. PMID- 11410017 TI - Antioxidative activities of phenylethanoid glycosides from Ligustrum purpurascens. AB - Tea and kudingcha (bitter tea) are the two most popular beverages consumed in China. Tea derived from the leaves of Camellia sinensis has been well studied for its various health benefits, but there are very limited data on the biological activities of bitter tea derived from the leaves of Ligustrum purpurascens (LP). The present study was carried out to characterize the antioxidants present in the bitter tea brewed from the leaves of LP. It was found that the crude glycoside fraction possessed strong protection against oxidation of human low-density lipoprotein (LDL). The column chromatographic separation led to the isolatation of five phenylethanoid glycosides, namely, acteoside, ligupurpuroside A, cis ligupurpuroside B, trans-ligupurpuroside B, and osmanthuside B. When acteoside was heated in the boiling water, it was isomerized to form isoacteoside. Acteoside, isoacteoside, and ligupurpuroside A purified from LP were protective, whereas cis-ligupurpuroside B, trans-ligupurpuroside B, and osmanthuside B exhibited no protection to human LDL from Cu(2+)-medicated oxidation. Acteoside, isoacteoside, and ligupurpuroside A were also effective in preventing the peroxyl free radical-induced oxidation of alpha-tocopherol in human LDL. The antioxidant activities of acteoside, isoacteoside, and ligupurpuroside A were comparable to that observed for a green tea antioxidant, (-)-epicatechin gallate. The inhibitory effect of these three phenylethanoid glycosides on oxidation of human LDL and alpha-tocopherol was dose-dependent at concentrations of 5-40 microM. The present results suggest that the bitter tea beverage derived from LP contains effective antioxidants that may have an equal benefit as a green tea beverage. PMID- 11410018 TI - Fate of fumonisins during the production of fried tortilla chips. AB - The fate of fumonisin B(1) (FB(1)), a mycotoxin found in corn, during the commercial manufacture of fried tortilla chips was studied. FB(1) and hydrolyzed FB(1) (HFB(1)) concentrations in four lots of corn and in the masa, other intermediates, liquid and waste byproducts, and fried chips were determined by HPLC. FB(1) concentrations in the masa and chips were reduced significantly, up to 80% in the fried chips, compared to that in the raw corn. HFB(1) was also found in the masa and chips, but at low concentrations compared to FB(1). LC-MS analyses corroborated HPLC findings and further showed the presence of partially hydrolyzed FB(1) (PHFB(1)), which, like HFB(1), was formed during the nixtamalization (cooking/steeping the corn in alkaline water to make masa) step and found predominantly in the cooking/steeping liquid and solid waste. No significant amounts of N-(carboxymethyl)-FB(1) or N-(1-deoxy-D-fructos-1-yl) FB(1), indicative of fumonisin-sugar adduct formation, were found. Thus, FB(1) is removed from corn and diverted into liquid and waste byproducts during the commercial production of fried tortilla chips. Nixtamalization and rinsing are the critical steps, whereas grinding, sheeting, baking, and frying the masa had little effect. PMID- 11410019 TI - Postharvest treatments for the reduction of mancozeb in fresh apples. AB - The objective of this study was to determine the effectiveness of chlorine, chlorine dioxide, ozone, and hydrogen peroxyacetic acid (HPA) treatments on the degradation of mancozeb and ethylenethiourea (ETU) in apples. This study was based on model experiments at neutral pH and temperature. Fresh apples were treated with two different levels of mancozeb (1 and 10 microg/mL). Several of the treatments were effective in reducing or removing mancozeb and ETU residues on spiked apples. Mancozeb residues decreased 56-99% with chlorine and 36-87% with chlorine dioxide treatments. ETU was completely degraded by 500 ppm of calcium hypochlorite and 10 ppm of chlorine dioxide at a 1 ppm spike level. However, at a 10 ppm spike level, the effectiveness of ETU degradation was lower than observed at 1 ppm level. Mancozeb residues decreased 56-97% with ozone treatment. At 1 and 3 ppm of ozone, no ETU residue was detected at 1 ppm of spiked mancozeb after both 3 and 30 min. HPA was also effective in degrading the mancozeb residues, with 44-99% reduction depending on treatment time and HPA concentrations. ETU was completely degraded at 500 ppm of HPA after 30 min of reaction time. These treatments indicated good potential for the removal of pesticide residues on fruit and in processed products. PMID- 11410021 TI - k-resonant benzenoid systems and k-cycle resonant graphs. AB - A benzenoid system (or hexagonal system) H is said to be k-resonant if, for 1 < or = t < or = k, any t disjoint hexagons of H are mutually resonant; that is, there is a Kekule structure (or perfect matching) K of H such that each of the k hexagons is an K-alternating hexagon. A connected graph G is said to be k-cycle resonant if, for 1 < or = t < or = k, any t disjoint cycles in G are mutually resonant. The concept of k-resonant benzenoid systems is closely related to Clar's aromatic sextet theory, and the concept of k-cycle resonant graphs is a natural generalization of k-resonant benzenoid systems. Some necessary and sufficient conditions for a benzenoid system (respectively a graph) to be k resonant (respectively k-cycle resonant) have been established. In this paper, we will give a survey on investigations of k-resonant benzenoid systems and k-cycle resonant graphs. PMID- 11410022 TI - Molecular similarity for small species: refining the isoelectronic index. AB - This paper tests a chemical-similarity model against properties of gas-phase neutral triatomic and four-atom molecules. The model is a variant of the Diatomics-in-Molecules (DIM) picture, which considers a molecule to be the superposition of all diatomic molecules that could be formed from adjacent atoms in the molecule. The variant is that adjacent atoms are counted as a diatomic molecule only if they are bonded. The tests consist of investigating whether molecules with the same number of electrons, computed by the adjacent-DIM model, have data more similar than do molecules selected at random. The tests vindicate the model for the heat of atomization and for the equilibrium constant for formation, they agree with the model with lesser confidence for the entropy and the partition function, and they show that the model fails for the ionization potential. The model applies with most confidence to molecules with the more electronegative atoms from rows 2 or 3 (those of greatest interest in organic chemistry) and with lesser confidence otherwise. For these properties and these molecules, the model passes graphical and statistical tests at least as well as does the traditional isoelectronic model. Thus, this work refines what may be called the isoelectronic index. PMID- 11410023 TI - A criteria to classify biological activity of benzimidazoles from a model of structural similarity. AB - We have classified a set of 250 benzimidazoles using a criterion of structural similarity. This criterion has led us to several clusters, which keep a close relationship between the molecules belonging to each one of them and their pharmacological activity. To study the structural similarity we have built a mathematical space where chemical structures are pictured as vectors. A set of well-chosen descriptors was used as variables. These descriptors arise from graph theoretical studies and quantum mechanical calculations. Principal components analysis was employed to find the suitable dimension for the space. Finally, cluster analysis was performed to classify the set of molecules by similarity. A Euclidean metric was used as a similarity coefficient. PMID- 11410024 TI - Comparison of a neural net-based QSAR algorithm (PCANN) with Hologram- and multiple linear regression-based QSAR approaches: application to 1,4 dihydropyridine-based calcium channel antagonists. AB - A QSAR algorithm (PCANN) has been developed and applied to a set of calcium channel blockers which are of special interest because of their role in cardiac disease and also because many of them interact with P-glycoprotein, a membrane protein associated with multidrug resistance to anticancer agents. A database of 46 1,4-dihydropyridines with known Ca2+ channel binding affinities was employed for the present analysis. The QSAR algorithm can be summarized as follows: (1) a set of 90 graph theoretic and information theoretic descriptors representing various structural and topological characteristics was calculated for each of the 1,4-dihydropyridines and (2) principal component analysis (PCA) was used to compress these 90 into the eight best orthogonal composite descriptors for the database. These eight sufficed to explain 96% of the variance in the original descriptor set. (3) Two important empirical descriptors, the Leo-Hansch lipophilic constant and the Hammet electronic parameter, were added to the list of eight. (4) The 10 resulting descriptors were used as inputs to a back propagation neural network whose output was the predicted binding affinity. (5) The predictive ability of the network was assessed by cross-validation. A comparison of the present approach with two other QSAR approaches (multiple linear regression using the same variables and a Hologram QSAR model) is made and shows that the PCANN approach can yield better predictions, once the right network configuration is identified. The present approach (PCANN) may prove useful for rapid assessment of the potential for biological activity when dealing with large chemical libraries. PMID- 11410025 TI - Graph-theoretical analysis of tunneling electron transfer in large polycyclic aromatic hydrocarbon networks. AB - Effect of a single nitrogen atom substitution to a number of large polycyclic aromatic hydrocarbon (PAH) molecules was calculated systematically, and it was found that especially in parallelogram-type PAH abnormal electron transfer (called tunneling electron transfer, TET) was observed. That is, fairly large amount of pi-electron is withdrawn to an electronegative nitrogen atom from almost the farthest end of a conjugated aromatic hydrocarbon molecule, leaving almost no change in the interior of the molecule. This change can be simulated by the Kekule structure counting for subgraphs of the parent molecule. PMID- 11410026 TI - Aromaticity in transition metal oxide structures. AB - The concept of aromaticity is useful for understanding the properties of some polyoxometalates containing transition metals such as vanadium, molybdenum, and tungsten having structures based on metal macropolygons and macropolyhedra with M O-M edges. Thus, the aromatic macrocuboctahedral Keggin ions readily undergo one electron reductions to highly colored mixed-valence "blues" (e.g., molybdenum blue), whereas the macroicosahedral Silverton ions, M(IV)Mo12O42(8-) (M(IV) = Ce, Th, U), which, like cyclohexane, do not have vertex valence orbitals available for delocalization, do not undergo analogous reduction reactions. A macrohexagon of d1 vanadium(IV) atoms as V-O-V units has been imbedded into an electronically inactive borate matrix in the ion [V6B20O50H8](8-). The small beta unit for the V O-V interactions in this V6 macrohexagon leads to an unprecedented example of high spin aromaticity with a paramagnetism corresponding to four unpaired electrons per V6 unit in contrast to benzene, which is diamagnetic and hence exhibits low spin aromaticity. The M-O-M interactions in these aromatic metal oxides are closely related to the Cu-O-Cu interactions in the high critical temperature superconducting copper oxides which are essential to the electron transport in these systems. PMID- 11410027 TI - Distance-related indexes in the quantitative structure-property relationship modeling. AB - A comparative study of structure-boiling point modeling for a set of 180 acyclic and cyclic hydrocarbons (DS-180) and two of its subsets (one containing a selection of 76 acyclic and cyclic alkanes (DS-76), and the other containing 104 (DS-104) mono- and polycyclic butanes through octanes) using several known and novel distance-related indices is reported. The distance-related indices used were as follows: Wiener index, hyper-Wiener index, detour index, hyper-detour index, Harary index, Pasareti index, Verhalom index, Wiener-sum index, inverse Wiener-sum index and the product-form version of the Wiener index. Additional indices used were the total number of paths, the Hosoya Z index, the total walk count index, the number of carbon atoms, and the number of rings in the hydrocarbon. The best models for predicting the boiling points of 76, 104, and 180 acyclic and cyclic alkanes contain the natural logarithm of the cross products of the Hosoya and detour index and of the Pasareti index and the number of rings. This result extends earlier work by us and Rucker and Rucker on the use of the Wiener, detour, and Hosoya indices in modeling boiling points of alkanes and cycloalkanes. It also supports later work by Rucker and Rucker on the use of the descriptor combination for the same purpose. PMID- 11410028 TI - Wiener index extension by counting even/odd graph distances. AB - Chemical structures of organic compounds are characterized numerically by a variety of structural descriptors, one of the earliest and most widely used being the Wiener index W, derived from the interatomic distances in a molecular graph. Extensive use of such structural descriptors or topological indices has been made in drug design, screening of chemical databases, and similarity and diversity assessment. A new set of topological indices is introduced representing a partitioning of the Wiener index based on counts of even and odd molecular graph distances. These new indices are further generalized by weighting exponents which can be optimized during the quantitative structure-activity/-property relationship (QSAR/QSPR) modeling process. These novel topological indices are tested in QSPR models for the boiling temperature, molar heat capacity, standard Gibbs energy of formation, vaporization enthalpy, refractive index, and density of alkanes. In many cases, the even/odd distance indices proposed here give notably improved correlations. PMID- 11410029 TI - On interpretation of well-known topological indices. AB - Many topological indices lack an interpretation in terms of simple physicochemical quantities. We have reexamined the structural interpretation of well-known topological indices: the connectivity index (1)chi, the Wiener index W, and the Hosoya topological index Z. We relate the success of various indices in structure-property studies to the degree to which they differentiate contributions from more exposed terminal bonds and more buried interior bonds. When considering bond additive properties of alkanes we find better regressions when greater weights are assigned to terminal CC bonds and lesser weights to internal CC. We suggest here that topological indices be discussed in terms of their partitioning into bond contributions, which for different indices and different bonds will assume different values. With this insight we modified the Wiener index W into a new index W, in which bond contributions are determined using the reciprocal of the product of the number of atoms on each side of a bond. Similarly we modified the Hosoya index Z into Z in which the frequency of occurrence of individual CC bonds in the patterns of disjoint bonds are considered. Novel indices are compared with other indices and they show to yield better regressions for boiling points of octane isomers. This suggests a useful classification of topological indices based on the relative magnitudes of the contributions of terminal and interior bonds. To extend such considerations to other indices one needs to consider partitioning of global molecular indices into bond additive terms. A general scheme for partitioning of molecular descriptors into bond contributions is outlined for indices derived from a selection of matrices associated with molecular graphs. PMID- 11410030 TI - Characterization of DNA primary sequences based on the average distances between bases. AB - We outline numerical characterization of DNA primary sequence based on calculation of the average distance between pairs of nucleic acid bases. This leads to a representation of DNA by a condensed 4 x 4 symmetrical matrix, the elements of which give the average separation between pair of bases X, Y in DNA (X, Y = A, C, G, T). As an invariant of choice we consider the leading eigenvalue of the derived 4 x 4 matrix. Additional structurally related invariants were obtained by constructing additional "higher order" 4 x 4 matrices derived from the initial 4 x 4 matrix by raising its elements to higher powers. Suitably normalized leading eigenvalue of these matrices offer a novel characterization of DNA primary sequences, referred to as "DNA profiles". The approach is illustrated on exon 1 of human beta-globin gene. PMID- 11410031 TI - Codessa-based theoretical QSPR model for hydantoin HPLC-RT lipophilicities. AB - A quantitative structure property relationship investigation was performed on the lipophilicities of a number of hydantoin derivatives as measured by the RP-HPLC retention times provided by Scholl et al. (Scholl, S.; Koch, A.; Henning, D.; Kempter, G.; Kleinpeter, E. Struct. Chem. 1999, 10, 355-366). The lipophilicities (S) were correlated with the theoretical molecular descriptors of the hydantoins obtained using the CODESSA program from the AM1-optimized geometry and electron wave functions. This study discloses enhanced correlations of the lipophilicities with the molecular descriptors, wherein the influence of the entropy factor is found to predominate. PMID- 11410032 TI - The variable molecular descriptors based on distance related matrices. AB - Recently variable molecular connectivity index and variable paths have been tested as molecular descriptors in several structure-property regressions. Here we outline the construction of several variable molecular descriptors, derived from the distance matrix and the "reversed" distance matrix. This includes the variable Balaban J index and the "reversed" Balaban index 1/J as well as a novel index 1/JJ derived from J and 1/J. All the variable descriptors mentioned were constructed by augmenting the distance matrix by replacing the diagonal zeroes with the variables x, y, z,. PMID- 11410033 TI - The overall Wiener index--a new tool for characterization of molecular topology. AB - Recently, the concept of overall connectivity of a graph G, TC(G), was introduced as the sum of vertex degrees of all subgraphs of G. The approach of more detailed characterization of molecular topology by accounting for all substructures is extended here to the concept of overall distance OW(G) of a graph G, defined as the sum of distances in all subgraphs of G, as well as the sum of eth-order terms, (e)OW(G), with e being the number of edges in the subgraph. Analytical expressions are presented for OW(G) of several basic classes of graphs. The overall distance is analyzed as a measure of topological complexity in acyclic and cyclic structures. The potential usefulness of the components of this generalized Wiener index in QSPR/QSAR is evaluated by its correlation with a number of properties of C3-C8 alkanes and by a favorable comparison with models based on molecular connectivity indices. PMID- 11410034 TI - On structural interpretation of several distance related topological indices. AB - We consider the role that individual bonds play in bond-additivities in order to better understand the structural basis of various topological indices. In particular we consider indices closely related to the Wiener index (W) and the distance matrix and search for optimal weights of terminal and interior CC bonds in alkanes for a selection of physicochemical properties. It is interesting to note that different properties are associated with different relative roles of the exterior and the interior CC bonds. PMID- 11410035 TI - Retro-regression--another important multivariate regression improvement. AB - We review the serious problem associated with instabilities of the coefficients of regression equations, referred to as the MRA (multivariate regression analysis) "nightmare of the first kind". This is manifested when in a stepwise regression a descriptor is included or excluded from a regression. The consequence is an unpredictable change of the coefficients of the descriptors that remain in the regression equation. We follow with consideration of an even more serious problem, referred to as the MRA "nightmare of the second kind", arising when optimal descriptors are selected from a large pool of descriptors. This process typically causes at different steps of the stepwise regression a replacement of several previously used descriptors by new ones. We describe a procedure that resolves these difficulties. The approach is illustrated on boiling points of nonanes which are considered (1) by using an ordered connectivity basis; (2) by using an ordering resulting from application of greedy algorithm; and (3) by using an ordering derived from an exhaustive search for optimal descriptors. A novel variant of multiple regression analysis, called retro-regression (RR), is outlined showing how it resolves the ambiguities associated with both "nightmares" of the first and the second kind of MRA. PMID- 11410036 TI - Novel shape descriptors for molecular graphs. AB - We report on novel graph theoretical indices which are sensitive to the shapes of molecular graphs. In contrast to the Kier's kappa shape indices which were based on a comparison of a molecular graph with graphs representing the extreme shapes, the linear graph and the "star" graph, the new shape indices are obtained by considering for all atoms the number of paths and the number of walks within a graph and then making the quotients of the number of paths and the number of walks the same length. The new shape indices show much higher discrimination among isomers when compared to the kappa shape indices. We report the new shape indices for smaller alkanes and several cyclic structures and illustrate their use in structure-property correlations. The new indices offer regressions of high quality for diverse physicochemical properties of octanes. They also have lead to a novel classification of physicochemical properties of alkanes. PMID- 11410037 TI - On use of the variable connectivity index 1chi(f) in QSAR: toxicity of aliphatic ethers. AB - The number of non-hydrogen atoms in a molecule, N, appears as a very good molecular descriptor for the toxicity of aliphatic ethers, despite the fact that it does not differentiate among isomers. The regression based on N as a descriptor for the toxicity in mice of 21 alkyl ethers was reported to yield the regression coefficient r = 0.9751. The simple connectivity index 1chi produced for the same data a less satisfactory regression: r = 0.9548. To see if variable connectivity index 1chi(f) can improve the regression characterized by N we examined the same data using the variable connectivity index 1chi(f) as a molecular descriptor. By varying x, y, the variables that discriminate between carbon and oxygen atoms, we obtained regression which approaches in quality the best reported regression using weighted paths as descriptor and which is marginally better than the regression based on N. PMID- 11410038 TI - On the characterization of DNA primary sequences by triplet of nucleic acid bases. AB - We consider construction of a set of smaller 4 x 4 matrices to represent DNA primary sequences which are based on enumeration of all 64 triplets of nucleic acids bases. The leading eigenvalue from the constructed matrices has been selected as an invariant for construction of a vector to characterize DNA. Additional invariants considered of the derived condensed matrices of DNA include a 64-component vector, the components of which consist of ordered triplets XYZ, with X, Y, Z = A, C, G, T. Construction of similarity/dissimilarity tables based on different invariants for a set of sequences of DNA belonging to the first exon of the beta-globin gene of eight species illustrates the utility of newly formulated invariants for DNA. PMID- 11410039 TI - Graph valence shells as molecular descriptors. AB - We have introduced a new simple structural descriptor for molecules that is based on the count of the valence shells for vertices in molecular graphs. The construction of the new descriptor is illustrated on 2,3-dimethylhexane and is reported for the 18 octane isomers. The relationship of the new descriptor to the path numbers of a graph is discussed. It can be seen that the path counts and the count of valence of neighbor shells are related for paths of length two (and shells of range two). There is no appreciable correlation between the count of the longer paths and the count of the corresponding neighbor valence shells at larger separations. Use of the neighbor valence shells as molecular descriptors is illustrated on the boiling point, the entropy, and the density of octanes. An intriguing situation is observed for regressions involving considered properties of n-octane isomers C8H18 in that the paths of length two, three, and four and the shells of the range two, three, and four give identical multivariate regression statistics. An explanation for this somewhat unusual aspect of MRA (multiple regression analysis) is offered. PMID- 11410040 TI - The variable connectivity index 1chi(f) versus the traditional molecular descriptors: a comparative study of 1chi(f) against descriptors of CODESSA. AB - In this study we compared the prediction abilities of the variable connectivity index 1chi(f) (not included in CODESSA) with topological indices available from CODESSA. We selected the boiling points of n = 100 alcohols as the property and examined the pool of 56 topological indices. Prediction capabilities of the developed models were evaluated by classical training/test set approach. RMS errors calculated from the prediction set for the MLR models obtained from CODESSA software with 1, 2, 3, 4, and 5 parameters were 9.06, 5.69, 5.40, 4.9, and 3.37 degrees C, respectively. Using the variable connectivity index with weights x = 0.10 and y = -0.92 for carbon and oxygen atom respectively, we obtain regression BP = 38.12 1chi(f) - 37.56 with the correlation coefficient r = 0.9915, RMS error 4.21 degrees C calculated from the test set, and Fisher ratio F = 5691. Prediction capability of the variable connectivity index was better than for MLR regression model with up to four parameters. PMID- 11410041 TI - Graph theoretical descriptors of two-dimensional chirality with possible extension to three-dimensional chirality. AB - We report on construction of a graph theoretical descriptor of chirality for two dimensional benzenoid hydrocarbons. For achiral molecules the index is zero, while for chiral molecules it takes a positive or negative value for a molecule and one for its mirror image. Hence, we also have obtained a descriptor that measures chirality on an absolute scale, i.e., it assigns to a molecule a positive (or a negative) value without ever requiring information on the enantiomer of the molecule considered or any other pair of standards. In addition, we have briefly outlined how this is the 2-D approach extended to characterization of chiral objects in 3-D space, at least in the case of n-alkane rotamers. PMID- 11410042 TI - A new descriptor for structure-property and structure-activity correlations. AB - We consider an improvement of a multiple regression analysis (MRA) for the correlation of boiling points of alcohols using descriptors that involve a variable part. In the search for the best descriptors based on weighted paths we came upon a novel molecular descriptor, the use of which was apparently overlooked in the past. The novel descriptor counts paths of length three; however, only those associated with an oxygen atom are counted. PMID- 11410043 TI - Variable connectivity index for cycle-containing structures. AB - In the early applications of the connectivity index the index was empirically modified for some properties of cyclic structures by subtracting 1/2 from the computed value based solely on valence of vertices in the molecular graph. In this article we looked into the origin of this heuristic adjustment of the connectivity indices for cyclic structures. We have examined the relative role of carbon atoms in cycle-containing structures by differentiating carbon atoms making up a ring and carbon atoms in exocyclic bonds. We found in the case of the boiling points of cycloalkanes and alkylcycloalkanes that contributions of "cyclic" and "acyclic" atoms to molecular additivities differ somewhat. PMID- 11410044 TI - QSAR with few compounds and many features. AB - Fitting quantitative structure-activity relationships (QSAR) requires different statistical methodologies and, to some degree, philosophies depending on the "shape" of the data matrix. When few features are used and there are many compounds, it is a reasonable expectation that good feature subset selection may be made and that nonlinearities and nonadditivities can be detected and diagnosed. Where there are many features and few compounds, this is unrealistic. Methods such as ridge regression RR, PLS, and principal component regression PCR, which abjure feature selection and rely on linearity may provide good predictions and fair understanding. We report a development of ridge regression for the underdetermined case by using generalized cross-validation to choose the ridge constant and perform F-tests for additional information. Conventional regression diagnostics can be used in followup to identify nonlinearities and other departures from model. We illustrate the approach with QSAR models of four data sets using calculated molecular descriptors. PMID- 11410045 TI - Prediction of mutagenicity of aromatic and heteroaromatic amines from structure: a hierarchical QSAR approach. AB - Due to the lack of experimental data, there has been increasing use of theoretical structural descriptors in the hazard assessment of chemicals. We have used a hierarchical approach to develop class-specific quantitative structure activity relationship (QSAR) models for the prediction of mutagenicity of a set of 95 aromatic and heteroaromatic amines. The hierarchical approach begins with the simplest molecular descriptors, the topostructural, which encode limited chemical information. The complexity is then increased, adding topochemical, geometric, and finally quantum chemical parameters. We have also added log P to the set of independent variables. The results indicate that the topological parameters, i.e., the topostructural and topochemical indices, explain the majority of the variance, and that the inclusion of log P, geometric, and quantum chemical parameters does not result in significantly improved predictive models. PMID- 11410046 TI - Interpretation of quantitative structure-property and -activity relationships. AB - The potential utility of data reduction methods (e.g. principal component analysis) for the analysis of matrices assembled from the related properties of large sets of compounds is discussed by reference to results obtained from solvent polarity scales, ongoing work on solubilities and sweetness properties, and proposed general treatments of toxicities and gas chromatographic retention indices. PMID- 11410047 TI - Further developments in determining the number of resonance structures in benzenoid free radicals: analytical expressions and elementary substructures. AB - Relationships for counting the number of resonance structures (SC) of benzenoid free radicals are discussed. The structural origin of some of the coefficients to the SC analytical expressions for series of benzenoid free radicals generated by successive attachment of specific aufbau units is determined. It is shown that a proper pruning fragment for the purpose of a priori determination of the coefficient in the highest degree n term of the SC analytical expression belonging to a given radical series has the property of bond fixation when the operation is performed on every member of that series, resulting in all remnant structures having the same SC value. PMID- 11410048 TI - Quantitative structure-property relationships (QSPRs) for the estimation of vapor pressure: a hierarchical approach using mathematical structural descriptors. AB - A set of 379 molecular descriptors was calculated for use in hierarchical quantitative structure-property relationship (QSPR) modeling of vapor pressure for a structurally diverse database consisting of 469 chemicals. The hierarchical approach utilizes topostructural, topochemical, geometrical, and quantum chemical descriptors in a stepwise fashion to develop QSPR models. In this way, the relative roles of the various levels of descriptors can be examined. The results show that the easily calculated topological descriptors explain the majority of the variance and that the addition of geometrical and quantum chemical descriptors does not result in a significantly improved model. PMID- 11410049 TI - Comparison of the NCI open database with seven large chemical structural databases. AB - Eight large chemical databases have been analyzed and compared to each other. Central to this comparison is the open National Cancer Institute (NCI) database, consisting of approximately 250 000 structures. The other databases analyzed are the Available Chemicals Directory ("ACD," from MDL, release 1.99, 3D-version); the ChemACX ("ACX," from CamSoft, Version 4.5); the Maybridge Catalog and the Asinex database (both as distributed by CamSoft as part of ChemInfo 4.5); the Sigma-Aldrich Catalog (CD-ROM, 1999 Version); the World Drug Index ("WDI," Derwent, version 1999.03); and the organic part of the Cambridge Crystallographic Database ("CSD," from Cambridge Crystallographic Data Center, 1999 Version 5.18). The database properties analyzed are internal duplication rates; compounds unique to each database; cumulative occurrence of compounds in an increasing number of databases; overlap of identical compounds between two databases; similarity overlap; diversity; and others. The crystallographic database CSD and the WDI show somewhat less overlap with the other databases than those with each other. In particular the collections of commercial compounds and compilations of vendor catalogs have a substantial degree of overlap among each other. Still, no database is completely a subset of any other, and each appears to have its own niche and thus "raison d'etre". The NCI database has by far the highest number of compounds that are unique to it. Approximately 200 000 of the NCI structures were not found in any of the other analyzed databases. PMID- 11410050 TI - Neural network based chemical structure indexing. AB - Searches on chemical databases are presently dominated by the text-based content of a paper which can be indexed into a keyword searchable form. Such traditional searches can prove to be very time-consuming and discouraging to the less frequent scientist. We report a simple chemical indexing based on the molecular structure alone. The method used is based on a one-to-one correspondence between the chemical structure presented as an image to a neural network and the corresponding binary output. The method is direct and less cumbersome (compared with traditional methods) and proves to be robust, elegant, and very versatile. PMID- 11410051 TI - Classification of environmental estrogens by physicochemical properties using principal component analysis and hierarchical cluster analysis. AB - A structurally diverse assortment of 60 environmental estrogens was divided into two main clusters ("A", "B") and a pair of subclusters ("C1", "C2") by applying principal component analysis to selected 1D and 2D molecular descriptors and subjecting the PCs to hierarchical cluster analysis. Although clustering was predicated solely on physicochemical properties, the dependence on particular physicochemical parameters of xenoestrogen binding affinities (pK(i)) to murine uterine cytosolic estrogen receptor (ER) proved greater for compounds within (sub)clusters than for compounds between (sub)clusters. Quantitative structure binding affinity relationships derived using molecular descriptors and PCs suggested differences in the driving forces for xenoestrogen-ER binding for different (sub)clusters. The modeling power for xenoestrogen-ER binding affinities of a combination of TLSER and WHIM 3D indices was much greater than that of combinations of 1D and 2D molecular descriptors or the PCs derived therefrom. The clusterings obtained using PCs also proved applicable to the 3D QSARs. PMID- 11410052 TI - Infrared spectra as chemical descriptors for QSAR models. AB - Infrared spectra (IR) were used as regressors for a number of QSARs and compared with both mechanistically oriented descriptors and heuristic "chemically neutral" descriptors (modified adjacency matrices eigenvalues). IR spectra usually gave results inferior to those obtained with the mechanistically driven descriptors, with one notable exception, and comparable to those obtained by adjacency matrices eigenvalues. So the IR spectra cannot be considered as an "a-priori" optimal description of molecules for QSAR. However the relation of IR with the chemicophysical bases of drug-receptor interaction suggests the use of IR spectra for elucidating mechanistic details. PMID- 11410053 TI - A fast algorithm for searching for molecules containing a pharmacophore in very large virtual combinatorial libraries. AB - We present a new algorithm for identifying molecules that display a pharmacophore, or in general a structural motif, by efficiently constructing and screening huge virtual combinatorial libraries of diverse compounds. The uniqueness of this algorithm is its ability to build and screen libraries of ca. 10(18) 3D molecular conformations within a reasonable time scale, thereby increasing the chemical space that can be virtually screened by many orders of magnitude. The algorithm may be used to design new molecules that display a desired pharmacophore on predefined sets of chemical scaffolds. This is demonstrated herein by screening a library of backbone cyclic peptides to find candidate peptido- and proteinomimetics. PMID- 11410054 TI - On walks in molecular graphs. AB - Walks in molecular graphs and their counts for a long time have found applications in theoretical chemistry. These are based on the fact that the (i, j)-entry of the kth power of the adjacency matrix is equal to the number of walks starting at vertex i, ending at vertex j, and having length k. In recent papers (refs 13, 18, 19) the numbers of all walks of length k, called molecular walk counts, mwc(k), and their sum from k = 1 to k = n - 1, called total walk count, twc, were proposed as quantities suitable for QSPR studies and capable of measuring the complexity of organic molecules. We now establish a few general properties of mwc's and twc among which are the linear dependence between the mwc's and linear correlations between the mwc's and twc, the spectral decomposition of mwc's, and various connections between the walk counts and the eigenvalues and eigenvectors of the molecular graph. We also characterize the graphs possessing minimal and maximal walk counts. PMID- 11410055 TI - Fingerprint scaling increases the probability of identifying molecules with similar activity in virtual screening calculations. AB - Results of systematic virtual screening calculations using a structural key-type fingerprint are reported for compounds belonging to 14 activity classes added to randomly selected synthetic molecules. For each class, a fingerprint profile was calculated to monitor the relative occupancy of fingerprint bit positions. Consensus bit patterns were determined consisting of all bits that were always set on in compounds belonging to a specific activity class. In virtual screening calculations, scale factors were applied to each consensus bit position in fingerprints of query molecules. This technique, called "fingerprint scaling", effectively increases the weight of consensus bit positions in fingerprint comparisons. Although overall prediction accuracy was satisfactory using unscaled calculations, scaling significantly increased the number of correct predictions but only slightly increased the rate of false positives. These observations suggest that fingerprint scaling is an attractive approach to increase the probability of identifying molecules with similar activity by virtual screening. It requires the availability of a series of related compounds and can be easily applied to any keyed fingerprint representation that associates bit positions with specific molecular features. PMID- 11410056 TI - Comparison of knowledge-based and distance geometry approaches for generation of molecular conformations. AB - A knowledge-based approach for generating conformations of molecules has been developed. The method described here provides a good sampling of the molecule's conformational space by restricting the generated conformations to those consistent with the reference database. The present approach, internally named et for enumerate torsions, differs from previous database-mining approaches by employing a library of much larger substructures while treating open chains, rings, and combinations of chains and rings in the same manner. In addition to knowledge in the form of observed torsion angles, some knowledge from the medicinal chemist is captured in the form of which substructures are identified. The knowledge-based approach is compared to Blaney et al.'s distance geometry (DG) algorithm for sampling the conformational space of molecules. The structures of 113 protein-bound molecules, determined by X-ray crystallography, were used to compare the methods. The present knowledge-based approach (i) generates conformations closer to the experimentally determined conformation, (ii) generates them sooner, and (iii) is significantly faster than the DG method. PMID- 11410057 TI - Quantum molecular similarity. 3. QTMS descriptors. AB - Building on the ideas of a previous paper [part 1, J. Phys. Chem. A 1999, 103, 2883] we present a new molecular similarity method based on the topology of the electron density. This method is directly applicable to QSARs and is called quantum topological molecular similarity (QTMS). It has been tested for five sets of carboxylic systems including para- and meta-benzoic acid, para-phenylacetic acid, 4-X-bicyclo[2.2.2]octane-1-carboxylic acids, and polysubstituted benzoic acids. In combination with the partial least squares (PLS) procedure QTMS is able to produce excellent and statistically valid regressions. It is shown that QTMS avoids certain challenges of traditional Carbo-like similarity indices. Finally, QTMS is able to suggest a molecular fragment that contains the active center or the part of the molecule that is responsible for the QSAR. PMID- 11410058 TI - Application of neural networks to modeling and estimating temperature-dependent liquid viscosity of organic compounds. AB - Back-propagation neural network models for correlating and predicting the viscosity-temperature behavior of a large variety of organic liquids were developed. Experimental values for the liquid viscosity for 1229 data points from 440 compounds containing C, H, N, O, S, and all halogens have been collected from the literature. The data ranges covered are from -120 to 160 degrees C for temperature and from 0.164 (trans-2-pentene at 20 degrees C) to 1.34 x 10(5) (glycerol at -20 degrees C) mPa.s for viscosity value. After dividing the total database of 440 compounds into training (237 with 673 data points), validation (124 with 423 data points), and test (79 with 133 data points) sets, the modeling performance of two separate neural network models with different architectures, one based on a compound-specific temperature dependence and the second based on a compound-independent one, has been examined. The resulting former model showed somewhat better modeling performance than latter, and the model gave squared correlation coefficients of 0.956, 0.932, and 0.884 and root mean-squares errors of 0.122, 0.134, and 0.148 log units for the training, validation, and test sets, respectively. The input descriptors include molar refraction, critical temperature, molar magnetic susceptibility, cohesive energy, temperatures, and five kinds of indicator variables for functionalities, alcohols/phenols, nitriles, amines, amides, and aliphatic ring. The reliability of the proposed model was assessed by comparing the results against calculated viscosities by two existing group-contribution approaches, the method of van Velzen et al. and the Joback and Reid method. PMID- 11410059 TI - 3D connectivity indices in QSPR/QSAR studies. AB - Topographic (3D) molecular connectivity indices based on molecular graphs weighted with quantum chemical parameters are used in QSPR and QSAR studies. These descriptors were compared to 2D connectivity indices (vertex and edge ones) and to quantum chemical descriptors in modeling partition coefficient (log P) and antibacterial activity of 2-furylethylene derivatives. In describing log P the 3D connectivity indices produced a significant improvement (more than 29%) in the predictive capacity of the model compared to those derived with topological and quantum chemical descriptors. The best linear discriminant model for classifying antibacterial activity of these compounds was also obtained with the use of 3D connectivity indices. The global percent of good classification obtained with 3D and 2D connectivity as well as quantum chemical descriptors were 94.1, 91.2, and 88.2, respectively. In general, all these models predict correctly the antibacterial activity of a set of nine new 2-furylethylene derivatives. The best result is obtained with 3D connectivity indices that classified correctly 100% of these compounds versus 88.9% obtained with 2D connectivity or quantum chemical descriptors. PMID- 11410060 TI - Design and prioritization of plates for high-throughput screening. AB - A general algorithm for the prioritization and selection of plates for high throughput screening is presented. The method uses a simulated annealing algorithm to search through the space of plate combinations for the one that maximizes some user-defined objective function. The algorithm is robust and convergent, and permits the simultaneous optimization of multiple design objectives, including molecular diversity, similarity to known actives, predicted activity or binding affinity, and many others. It is shown that the arrangement of compounds among the plates may have important consequences on the ability to design a well-targeted and cost-effective experiment. To that end, two simple and effective schemes for the construction of homogeneous and heterogeneous plates are outlined, using a novel similarity sorting algorithm based on one-dimensional nonlinear mapping. PMID- 11410061 TI - Construction of a perfectly N-representable two-electron density matrix carrying full information on an interacting system. AB - The two-particle density matrix contains all information, which is "readable" by one- and two-particle operators. The report discusses the building of a two electron density matrix, which is perfectly N-representable by construction. The 2-matrix of a pure quantum state is obtained without the intermediate determination of the wave function. The procedure is illustrated by computations on atomic and small molecular systems. PMID- 11410062 TI - Prediction of nonlinear optical responses of organic compounds. AB - The nonlinear optical quantities, second and third harmonics (beta and gamma), were predicted using a quantitative structure-property relationship (QSPR) approach. Molecular orbital ab initio calculations were applied to generate easily accessible variables to be used in the partial least-squares analysis. Simplified equations are presented that could be used to predict the experimental beta and gamma responses, prior to further investigations of potentially interesting molecules for use in optical materials. PMID- 11410063 TI - Association of two 3D QSAR analyses. application to the study of partial agonist serotonin-3 ligands. AB - CATALYST and COMFA, two software packages for 3D QSAR studies, were associated to correlate the three-dimensional structures of 75 serotonin 5-HT3 ligands to their biological affinities. The conformational analysis and the influence of chemical function-based alignments (the basis of this association) on final results are discussed in this publication. These two analyses allow for precisely quantitating the weights of significant chemical groups or functions on the biological affinities. PMID- 11410064 TI - Optimization and empirical modeling of HG-ICP-AES analytical technique through artificial neural networks. AB - An artificial neural network technique has been applied to the optimization of a hydride generation-inductively coupled plasma-atomic emission spectrometry (HG ICP-AES) coupling for the determination of Ge at trace levels. The back propagation of errors net architecture was used. Experimental parameters and their relationship have been studied, obtaining a surface response of the system. The results and optimization aspects achieved with the neural network approach have been compared to the "one variable at time" and SIMPLEX methods. PMID- 11410065 TI - Quantitative structure-activity relationship studies using Gaussian processes. AB - A Gaussian process method (GPM) is described and applied to the production of some QSAR models. These models have the potential to solve a number of problems which arise in QSAR modeling in that no parameters have to be supplied and only one hyperparameter is used in finding the optimal solution. The application of the method to QSAR is illustrated using data sets of compounds active at the benzodiazepine and muscarinic receptors as well as the data set of the toxicity of substituted benzenes to the ciliate, Tetrahymena Pyriformis. PMID- 11410066 TI - How far are molecular connectivity descriptors from I(S) molecular pseudoconnectivity descriptors? AB - A comparison of the characteristics of the molecular connectivity and intrinsic state pseudoconnectivity indices in modeling different activities and properties of different classes of compounds is performed. Two different activities of chlorofluorocarbons, an activity of 2-Br-2-phenetylamines, an activity of benzimidazoles, and the boiling points of primary amines and primary alcohols are modeled. The simultaneous modeling of these last two classes of compounds is also accomplished. The comparison encompasses also the molar mass as descriptor. Further, the influence of the rescaling procedure on the modeling for the case of benzimidazoles is analyzed as well as the relationship between degeneracy of connectivity values and quality of the modeling. Molecular pseudoconnectivity terms seem to compete advantageously with molecular connectivity terms in the descriptive power of the activity and property of many classes of compounds. In many cases new mixed higher-order connectivity-pseudoconnectivity terms have been detected that consistently improve the quality of the description. PMID- 11410067 TI - 3D-quantitative structure-activity relationships of HEPT derivatives as HIV-1 reverse transcriptase inhibitors, based on Ab initio calculations. AB - Comparative molecular field analysis (CoMFA) has been applied to a large set of 1 [(2-hydroxyethoxy)methyl]-6-(phenylthio)thymine (HEPT) analogues. The starting geometry of HEPT was obtained from crystallographic data of HEPT/HIV-1 reverse transcriptase (RT) complexes. The structures of 101 HEPT derivatives were considered and fully optimized by ab initio molecular orbital calculations at the HF/3-21G level. The best CoMFA model is satisfactory in both statistical significance and predictive ability. It shows excellent, high predictive ability as r2cv = 0.858. The derived model indicates the importance of steric contributions (64.4%) as well as electrostatic interactions for the HIV-1 RT inhibition. In addition, steric and electrostatic contour maps from this analysis agree well with the experimentally observed trend that there are steric interactions between the side chain of HEPT and an aromatic ring of Tyr181. It is concluded that a moderately sized group at C5 enhances contact with Tyr181 enough to push it into a position which renders the protein nonfunctional, but a smaller group has insufficient steric requirements to do this and a larger group renders the ligand too large for the cavity. The mutation-induced resistance of reverse transcriptase is explained by this analysis. The obtained results not only lead to a better understanding of structural requirements of this set of compounds for the inhibition but also enable the suggestions for new and more potent drugs. PMID- 11410068 TI - Molecular complexity and its impact on the probability of finding leads for drug discovery. AB - Using a simple model of ligand-receptor interactions, the interactions between ligands and receptors of varying complexities are studied and the probabilities of binding calculated. It is observed that as the systems become more complex the chance of observing a useful interaction for a randomly chosen ligand falls dramatically. The implications of this for the design of combinatorial libraries is explored. A large set of drug leads and optimized compounds is profiled using several different properties relevant to molecular recognition. The changes observed for these properties during the drug optimization phase support the hypothesis that less complex molecules are more common starting points for the discovery of drugs. An extreme example of the use of simple molecules for directed screening against thrombin is provided. PMID- 11410070 TI - Letter to the editor. PMID- 11410071 TI - Cardiovascular disease: C-reactive protein and the inflammatory disease paradigm: HMG-CoA reductase inhibitors, alpha-tocopherol, red yeast rice, and olive oil polyphenols. A review of the literature. AB - The current understanding of the origin of atherosclerosis is that of an inflammatory process that involves the acute phase response -an innate biological response to a disturbance in homeostasis -infection, inflammation, tissue injury, neoplasm, or immune disturbance. The activation of the acute phase response, signaled by interleukin-6, produces proteins (fibrinogen, C-reactive protein (CRP), serum amyloid A) that lead to inflammatory reactions. The tissues themselves contain elevated levels of acute phase proteins and cytokines resulting in a localized inflammatory effect. Localized inflammatory responses in the intimal layer of the arterial wall have been shown to be responsible for many of the aspects of intimal thickening and plaque disruption, leading to acute cardiovascular events. The predictive value of plasma C-reactive protein as a risk factor for cardiovascular events has led some researchers to support the use of CRP as a main cardiovascular risk assessment tool, along with total cholesterol:HDL ratios and homocysteine levels. The ability of HMG-CoA reductase inhibitors to lower C-reactive protein levels has recently brought into question the mechanisms of action of the statin drugs. Because these medications lower incidences of acute cardiovascular events as well as decreasing morbidity and mortality well before the effects of lowered LDL cholesterol can be expected to occur, questions have been asked about whether they may work independently of LDL lowering mechanisms. Red yeast rice contains a naturally-occurring statin (lovastatin) as well as other cholesterol-lowering compounds, some with antioxidant effects. Alpha-tocopherol also significantly lowers CRP levels in diabetics and nondiabetics, and minimizes other aspects of the acute phase response and inflammatory damage involved in atherosclerosis. This may account for alpha-tocopherol's positive effect on cardiovascular morbidity and mortality. Finally, polyphenolic compounds present in virgin olive oil also have anti inflammatory and antioxidative effects in cardiovascular disease. The phenolic compounds in virgin olive oil may explain some of the protective effects found in epidemiological studies. PMID- 11410072 TI - Polycystic ovary syndrome: clinical considerations. AB - Polycystic ovary syndrome (PCOS) is one of the most frequently encountered endocrine disorders occurring in women of reproductive age. Clinically, a patient usually presents with menstrual irregularities, infertility, and hirsutism. If not treated properly, a patient is at risk for type 2 diabetes, cardiovascular disease, and hyperestrogen-related cancers. The hallmark endocrine disorders of this syndrome are hyperandrogenism and hyperinsulinemia. Great controversy exists as to which state precedes the other. There also appears to be a defect in the hypothalamic-pituitary-adrenal (HPA) axis in patients presenting with polycystic ovary syndrome. Research consistently demonstrates that the first line of treatment for this condition is weight loss. Weight loss and dietary changes appear to affect all parameters of hormonal fluctuation. Due to the vast array of side effects associated with many pharmaceutical agents typically prescribed to treat PCOS, natural therapeutics including nutrient supplementation and botanicals may be a less invasive and equally effective approach. Due to the seriousness of this syndrome when left untreated, prompt evaluation and treatment is essential. PMID- 11410073 TI - Rhodiola rosea: a possible plant adaptogen. AB - Rhodiola rosea is a popular plant in traditional medical systems in Eastern Europe and Asian with a reputation for stimulating the nervous system, decreasing depression, enhancing work performance, eliminating fatigue, and preventing high altitude sickness. Rhodiola rosea has been categorized as an adaptogen by Russian researchers due to its observed ability to increase resistance to a variety of chemical, biological, and physical stressors. Its claimed benefits include antidepressant, anticancer, cardioprotective, and central nervous system enhancement. Research also indicates great utility in asthenic conditions (decline in work performance, sleep difficulties, poor appetite, irritability, hypertension, headaches, and fatigue) developing subsequent to intense physical or intellectual strain. The adaptogenic, cardiopulmonary protective, and central nervous system activities of Rhodiola rosea have been attributed primarily to its ability to influence levels and activity of monoamines and opioid peptides such as beta-endorphins. PMID- 11410074 TI - An extract of Petasites hybridus is effective in the prophylaxis of migraine. AB - OBJECTIVE: Migraine is still an unsolved problem. This clinical trial investigates the efficacy and tolerance of Petasites hybridus in the prophylaxis of migraine. METHODS: A randomized, group-parallel, placebo-controlled, double blind clinical study was carried out with a special CO2 extract from the rhizome of Petasites hybridus. Following a four-week run-in phase, 60 patients received either the special Petasites hybridus extract Petadolex or placebo at a dosage of two capsules (each capsule contains 25 mg) twice daily over 12 weeks. Outcome variables included the frequency, intensity and duration of migraine attacks as well as any accompanying symptoms. RESULTS: The frequency of migraine attacks decreased by a maximum of 60 percent compared to the baseline. This reduction in migraine attacks with Petadolex was significant (p < 0.05) compared to placebo. No adverse events were reported. Petasites was exceptionally well tolerated. CONCLUSIONS: The results suggest that migraine patients can benefit from prophylactic treatment with this special extract. The combination of high efficacy and excellent tolerance emphasizes the particular value that Petasites hybridus has for the prophylactic treatment of migraine. PMID- 11410075 TI - Transdermal secretin for autism - a case report. AB - Secretin hormone given daily in transdermal cream was associated with marked and sustained developmental progress in an aphasic two-and-a-half year old child diagnosed with autism. PMID- 11410076 TI - DHEA. Monograph. AB - Dehydroepiandrosterone (DHEA) is a steroid hormone secreted primarily by the adrenal glands and to a lesser extent by the brain, skin, testes, and ovaries. It is the most abundant circulating steroid in humans and can be converted into other hormones, including estrogen and testosterone. It has been characterized as a pleiotropic "buffer hormone," with receptor sites in the liver, kidney, and testes, and has a key role in a wide range of physiological responses. Circulating levels of DHEA decline with age and a relationship has been suggested between lower DHEA levels and heart disease, cancer, diabetes, obesity, chronic fatigue syndrome, AIDS, and Alzheimer's disease. Other research suggests that autoimmune diseases such as systemic lupus erythematosus (SLE), rheumatoid arthritis, and multiple sclerosis might be associated with declining DHEA levels. PMID- 11410077 TI - Picrorhiza kurroa. Monograph. AB - Picrorhiza kurroa is a well-known herb in the Ayurvedic system of medicine and has traditionally been used to treat disorders of the liver and upper respiratory tract, reduce fevers, and to treat dyspepsia, chronic diarrhea, and scorpion sting. It is a small perennial herb from the Scrophulariaceae family, found in the Himalayan region growing at elevations of 3,000 - 5,000 meters. Picrorhiza kurroa has a long, creeping rootstock that is bitter in taste, and grows in rock crevices and moist, sandy soil. The leaves of the plant are flat, oval, and sharply serrated. The flowers, which appear June through August, are white or pale purple and borne on a tall spike; manual harvesting of the plant takes place October through December. The active constituents are obtained from the root and rhizomes. The plant is self-regenerating but unregulated over-harvesting has caused it to be threatened to near extinction. Current research on Picrorhiza kurroa has focused on its hepatoprotective, anticholestatic, antioxidant, and immune-modulating activity. PMID- 11410079 TI - HIV/AIDS in the Caribbean: big problems among small islands. PMID- 11410080 TI - Pioneering work in several fields in Spain: Barcelona's municipal institute of biomedical research. PMID- 11410086 TI - Intensity of physical activity and risk of coronary heart disease. PMID- 11410087 TI - Intensity of physical activity and risk of coronary heart disease. PMID- 11410089 TI - Hormonal replacement therapy and cognition in postmenopausal women. PMID- 11410091 TI - Brassica vegetables and breast cancer risk. PMID- 11410093 TI - Histamine poisoning from seafood. PMID- 11410095 TI - Rates of fetal death by maternal race, ethnicity, and nativity: New Jersey, 1991 1998. PMID- 11410096 TI - Impact of folic acid fortification of the US food supply on the occurrence of neural tube defects. AB - CONTEXT: Daily consumption of 400 microg of folic acid before conception and during early pregnancy dramatically reduces the occurrence of neural tube defects (NTDs). Before food fortification, however, only an estimated 29% of US reproductive-aged women were taking a supplement containing 400 microg of folic acid daily. The US Food and Drug Administration authorized addition of folic acid to enriched grain products in March 1996, with compliance mandatory by January 1998. OBJECTIVE: To evaluate the impact of food fortification with folic acid on NTD birth prevalence. DESIGN, SETTING, AND POPULATION: National study of birth certificate data for live births to women in 45 US states and Washington, DC, between January 1990 and December 1999. MAIN OUTCOME MEASURE: Birth certificate reports of spina bifida and anencephaly before fortification (October 1995 through December 1996) compared with after mandatory fortification (October 1998 through December 1999). RESULTS: The birth prevalence of NTDs reported on birth certificates decreased from 37.8 per 100 000 live births before fortification to 30.5 per 100 000 live births conceived after mandatory folic acid fortification, representing a 19% decline (prevalence ratio [PR], 0.81; 95% confidence interval [CI], 0.75-0.87). During the same period, NTD birth prevalence declined from 53.4 per 100 000 to 46.5 per 100 000 (PR, 0.87; 95% CI, 0.64-1.18) for women who received only third-trimester or no prenatal care. CONCLUSIONS: A 19% reduction in NTD birth prevalence occurred following folic acid fortification of the US food supply. However, factors other than fortification may have contributed to this decline. PMID- 11410097 TI - Development and validation of a prognostic index for 1-year mortality in older adults after hospitalization. AB - CONTEXT: For many elderly patients, an acute medical illness requiring hospitalization is followed by a progressive decline, resulting in high rates of mortality in this population during the year following discharge. However, few prognostic indices have focused on predicting posthospital mortality in older adults. OBJECTIVE: To develop and validate a prognostic index for 1 year mortality of older adults after hospital discharge using information readily available at discharge. DESIGN: Data analyses derived from 2 prospective studies with 1-year of follow-up, conducted in 1993 through 1997. SETTING AND PATIENTS: We developed the prognostic index in 1495 patients aged at least 70 years who were discharged from a general medical service at a tertiary care hospital (mean age, 81 years; 67% female) and validated it in 1427 patients discharged from a separate community teaching hospital (mean age, 79 years; 61% female). MAIN OUTCOME MEASURE: Prediction of 1-year mortality using risk factors such as demographic characteristics, activities of daily living (ADL) dependency, comorbid conditions, length of hospital stay, and laboratory measurements. RESULTS: In the derivation cohort, 6 independent risk factors for mortality were identified and weighted using logistic regression: male sex (1 point); number of dependent ADLs at discharge (1-4 ADLs, 2 points; all 5 ADLs, 5 points); congestive heart failure (2 points); cancer (solitary, 3 points; metastatic, 8 points); creatinine level higher than 3.0 mg/dL (265 micromol/L) (2 points); and low albumin level (3.0-3.4 g/dL, 1 point; <3.0 g/dL, 2 points). Several variables associated with 1-year mortality in bivariable analyses, such as age and dementia, were not independently associated with mortality after adjustment for functional status. We calculated risk scores for patients by adding the points of each independent risk factor present. In the derivation cohort, 1-year mortality was 13% in the lowest-risk group (0-1 point), 20% in the group with 2 or 3 points, 37% in the group with 4 to 6 points, and 68% in the highest-risk group (>6 points). In the validation cohort, 1-year mortality was 4% in the lowest-risk group, 19% in the group with 2 or 3 points, 34% in the group with 4 to 6 points, and 64% in the highest-risk group. The area under the receiver operating characteristic curve for the point system was 0.75 in the derivation cohort and 0.79 in the validation cohort. CONCLUSIONS: Our prognostic index, which used 6 risk factors known at discharge and a simple additive point system to stratify medical patients 70 years or older according to 1-year mortality after hospitalization, had good discrimination and calibration and generalized well in an independent sample of patients at a different site. These characteristics suggest that our index may be useful for clinical care and risk adjustment. PMID- 11410098 TI - Risks for incident human papillomavirus infection and low-grade squamous intraepithelial lesion development in young females. AB - CONTEXT: Low-grade squamous intraepithelial lesions (LSILs) have been described as a benign cytological consequence of active human papillomavirus (HPV) replication. Several studies have reported that certain behavioral and biological risks exist for LSIL, suggesting that HPV alone is not sufficient for the development of LSIL. However, because most of these studies have been cross sectional, it is not known whether behavioral and biological risks are simply risks for HPV infection itself. OBJECTIVE: To prospectively examine risks of incident HPV infection in HPV-negative females and of incident LSIL development in females with HPV infection. DESIGN: Prospective cohort study conducted between 1990-2000, with a median follow-up of 50 months. SETTING AND PARTICIPANTS: Females aged 13 to 21 years who attended 2 family planning clinics in the San Francisco bay area; 496 had prevalent HPV infection and 105 were HPV-negative. MAIN OUTCOME MEASURE: Incident development of HPV infection and LSIL, analyzed by various demographic, behavioral, and clinical risk factors. RESULTS: Fifty-four incident HPV infections occurred in the 105 females who were HPV-negative at study entry (median duration of follow-up for those who remained HPV-negative was 26 months). Multivariable analysis showed that risks of HPV included sexual behavior (relative hazard [RH], 10.10; 95% confidence interval [CI], 3.24-31.50 per new partner per month), history of herpes simplex virus (RH, 3.54; 95% CI, 1.37-9.10), and history of vulvar warts (RH, 2.73; 95% CI, 1.27-5.87). Current use of oral contraceptives had a significantly protective effect (RH, 0.49; 95% CI, 0.28-0.86). Among the 496 individuals who were HPV-positive at baseline or in follow-up, there were 109 incident cases of LSIL during the follow-up interval, with a median follow-up time of 60 months for those who never developed LSIL. Human papillomavirus infection was the most significant risk factor for development of LSIL. The multivariable model showed the following risks for LSIL: HPV infection for less than 1 year (RH, 7.40; 95% CI, 4.74-11.57); HPV infection for 1 to 2 years (RH, 10.27; 95% CI, 5.64-18.69); HPV infection for 2 to 3 years (RH, 6.11; 95% CI, 1.86-20.06); and daily cigarette smoking (RH, 1.67; 95% CI, 1.12-2.48). CONCLUSION: Our results indicate distinct risks for HPV and LSIL. In addition, most women with HPV infection in our study did not develop LSIL within a median follow-up period of 60 months. These findings underscore the hypothesis that certain biological risks thought to be associated with LSIL are, in fact, risks for acquisition of HPV. Cigarette smoking was a risk specific to LSIL, supporting the role of tobacco in neoplastic development. PMID- 11410099 TI - Alabama coronary artery bypass grafting project: results of a statewide quality improvement initiative. AB - CONTEXT: Efforts to improve quality of care in the cardiac surgery field have focused on reducing the risk-adjusted mortality associated with common surgical procedures, such as coronary artery bypass grafting (CABG). However, the best methodological approach to improvement is under debate. OBJECTIVE: To test an intervention to improve performance of CABG surgery. DESIGN AND SETTING: Quality improvement project based on baseline (July 1, 1995-June 30, 1996) and follow-up (July 1-December 31, 1998) performance measurements from medical record review for all 20 Alabama hospitals that provided CABG surgery. PATIENTS: Medicare patients discharged after CABG surgery in Alabama (n = 5784), a comparison state (n = 3214), and a national sample (n = 3758). INTERVENTION: Confidential hospital specific performance feedback and assistance with multimodal improvement interventions, including the option to share relevant experience with peers. MAIN OUTCOME MEASURES: Duration of intubation, reintubation rate, aspirin therapy at discharge, use of the internal mammary artery (IMA), hospital readmission rate, and risk-adjusted in-hospital mortality. RESULTS: Proportion of extubation within 6 hours increased from 9% to 41% in Alabama, decreased from 40% to 39% in the comparison state, and increased from 12% to 25% in the national sample. Use of IMA increased from 73% to 84%, 48% to 55%, and 74% to 81%, respectively, in the 3 samples, but aspirin use increased only in Alabama (from 88% to 92%). The amount of improvement in all 3 of these process measures was greater in Alabama than in the other samples (IMA use for Alabama vs comparison state was P =.001 and for Alabama vs national sample, P =.02; and P<.001 for all other comparisons). Risk adjusted mortality decreased in Alabama (4.9% to 2.9%), but this decrease was not statistically significantly different from mortality changes in the other groups (odds ratio, 0.76; 95% confidence interval, 0.54-1.07 vs national sample). CONCLUSION: Confidential peer-based regional performance feedback and process oriented analysis of shared experience are associated with some improvement in quality of care for patients who underwent CABG surgery. PMID- 11410100 TI - Risk factors for and impact of online sexual solicitation of youth. AB - CONTEXT: Health care professionals, educators, and others are increasingly called upon to advise parents and policymakers about risks posed to children by Internet use. However, little scientific information exists on the experiences of children online. OBJECTIVE: To assess the risk factors surrounding online sexual solicitations of youth and distress due to solicitation. DESIGN, SETTING, AND PARTICIPANTS: Telephone survey (August 1999-February 2000) of a random sample of 1501 youth aged 10 through 17 years who were regular Internet users. MAIN OUTCOME MEASURES: Demographic and behavioral characteristics associated with solicitation risk and distress due to solicitation. RESULTS: Nineteen percent of youth who used the Internet regularly were the targets of unwanted sexual solicitation in the last year. Girls (P<.001), older teens (P =.005), troubled youth (P =.004), frequent Internet users (P =.01), chat room participants (P<.001), and those who communicated online with strangers (P<.001) were at greater risk. Twenty-five percent of the solicited youth reported high levels of distress after solicitation incidents. Risk of distress was more common among the younger youth (P =.005), those who received aggressive solicitations (the solicitor attempted or made offline contact) (P =.001), and those who were solicited on a computer away from their home (P =.001). CONCLUSIONS: Many young people who use the Internet encounter unwanted sexual overtures. Health care professionals, educators, and parents should be prepared to educate youth about how to respond to online sexual solicitations, including encouraging youth to disclose and report such encounters and to talk about them. PMID- 11410101 TI - National health information privacy: regulations under the Health Insurance Portability and Accountability Act. AB - Health information privacy is important in US society, but existing federal and state law does not offer adequate protection. The Department of Health and Human Services, under powers granted by the Health Insurance Portability and Accountability Act of 1996, recently issued a final rule providing systematic, nationwide health information privacy protection. The rule is extensive in its scope, applying to health plans, health care clearinghouses, and health care providers (hospitals, clinics, and health departments) who conduct financial transactions electronically ("covered entities"). The rule applies to personally identifiable information in any form, whether communicated electronically, on paper, or orally. The rule does not preempt state law that affords more stringent privacy protection; thus, the health care industry will have to comply with multiple layers of federal and state law. The rule affords patients rights to education about privacy safeguards, access to their medical records, and a process for correction of records. It also requires the patient's permission for disclosures of personal information. While privacy is an important value, it may conflict with public responsibilities to use data for social goods. The rule has special provisions for disclosure of health information for research, public health, law enforcement, and commercial marketing. The privacy debate will continue in Congress and within the president's administration. The primary focus will be on the costs and burdens on health care providers, the ability of health care professionals to use and share full medical information when treating patients, the provision of patient care in a timely and efficient manner, and parents' access to information about the health of their children. PMID- 11410102 TI - Food fortification to prevent neural tube defects: is it working? PMID- 11410103 TI - Prognostic indices in clinical practice. PMID- 11410108 TI - Maximizing the success rate of minimal model insulin sensitivity measurement in humans: the importance of basal glucose levels. AB - Minimal model analysis of glucose and insulin concentrations in the intravenous glucose tolerance test (IVGTT) has been widely used to obtain a measure of insulin sensitivity in humans. Issues of model validity and IVGTT protocol have been explored extensively. Less attention has been paid, however, to the computer programming protocol for estimating the model parameters (programming implementation). Minimal model analysis of data from an IVGTT protocol involving a high glucose dose (0.5 g/kg) and a reduced sample schedule, employed in healthy pre- or post-menopausal women, healthy men or men with coronary heart disease or chronic heart failure (20 in each group), was undertaken according to 12 different programming implementations using a commercially available model equation-solving program. The ability of the program to arrive at an acceptable solution to the model equations gave a success rate of between 39% and 96%, depending on the implementation. Variation in basal glucose assignment significantly affected the magnitude of estimates of insulin sensitivity. The maximum modelling success rate was achieved by introduction of an imputed glucose measurement at 360 min from the glucose injection, taking the basal glucose level as the fasting glucose concentration, and overweighting the initial glucose measurement after a delay for mixing. Use of this implementation to analyse data from a study comparing insulin sensitivities obtained using the minimal model and a euglycaemic clamp reference gave a correlation of 0.80 (P<0.001) between the two methods. Straightforward variations in programming implementation, involving appropriate assignment of the basal glucose concentration and use of an imputed glucose measurement signifying re-establishment of basal glucose levels following the IVGTT, can considerably improve modelling success rate. PMID- 11410109 TI - Bimodal effects of platelet-derived growth factor on rat mesangial cell proliferation and death, and the role of lysophosphatidic acid in cell survival. AB - Although mesangial cell death has been shown to be correlated with mesangial cell mitosis in vivo, little is known about how these two apparently opposite events are regulated. We show that the addition of platelet-derived growth factor (PDGF; 10-50 ng/ml) to primary cultured rat mesangial cells for 24 h caused continuous proliferation along with simultaneous cell death. This process was accompanied by the fragmentation of DNA into nucleosomal oligomers, the development of apoptotic morphological changes in the nucleus, and increased expression of p53. Accumulation of lactate dehydrogenase (LDH) was also observed in the culture medium, suggesting that both apoptosis and necrosis are involved in the cell death mechanisms observed. We also observed that addition of 30 microM lysophosphatidic acid (LPA) to the culture medium greatly suppressed PDGF-induced cell death, leading to synergistically enhanced mesangial cell proliferation. DNA fragmentation, p53 expression and LDH release were all suppressed by LPA. We suggest that PDGF is a bifunctional molecule in mesangial cells that evokes both cell proliferation and cell death simultaneously, whereas LPA is a survival factor. We speculate that PDGF and LPA may play important roles in the progression or exacerbation of proliferative glomerulonephritis. PMID- 11410110 TI - Adipose tissue thickness affects in vivo quantitative near-IR spectroscopy in human skeletal muscle. AB - The influence of adipose tissue thickness (ATT) on near-IR spectroscopy (NIRS) measurements in vivo was studied in the human flexor digitorum superficialis muscle at rest and during sustained isometric handgrip exercise. NIRS was used for the quantitative measurement of muscle O(2) consumption (mV.O(2)) and forearm blood flow (FBF) in 78 healthy subjects. Skinfold thickness ranged from 1.4 to 8.9 mm within the group. Resting mV.O(2) was 0.11+/-0.04 ml of O(2).min(-1).100 g(-1), and FBF was 1.28+/-0.82 ml.min(-1).100 ml(-1). There was a negative correlation (r=-0.70, P< or =0.01), indicating a decrease in mV.O(2) with increasing ATT. mV.O(2) in the 10 leanest subjects appeared to be twice as high as that in the 10 subjects with the highest ATT. A poor correlation (r=0.29, P< or =0.01) was found between ATT and FBF. The gender difference that we found for mV.O(2) was due to the difference in ATT between female and male subjects. No correlation was found between maximum voluntary contraction and mV.O(2), nor between maximum voluntary contraction and ATT, indicating that the contraction force did not confound our results. These results show that ATT has a substantial confounding influence on in vivo NIRS measurements, and that it is essential to incorporate this factor into future NIRS muscle studies in order to justify comparisons between different groups. To facilitate such comparisons, upper and lower boundaries for normal values of mV.O(2) and FBF in relation to ATT are presented. PMID- 11410111 TI - Is pseudocholinesterase activity related to markers of triacylglycerol synthesis in Type II diabetes mellitus? AB - Hypertriglyceridaemia is a risk factor for cardiovascular disease in patients suffering from Type II diabetes mellitus, and is due to enhanced synthesis and/or impaired clearance of triacylglycerol-rich lipoproteins. In the present study we investigated whether pseudocholinesterase (PChE) activity could serve as a marker for the rate of triacylglycerol synthesis in these patients. Patients were stratified according to their apolipoprotein E (apoE) phenotype, i.e. E3E2, E3E3 or E3E4. In study I, the relationship between PChE activity and serum triacylglycerols was investigated in 224 insulin-treated patients with Type II diabetes. In study II, which had a cross-over design, PChE activity was measured in 45 dyslipidaemic, insulin-treated patients with Type II diabetes that were treated with bezafibrate or pravastatin. In study I, PChE activity was correlated positively with serum triacylglycerol concentrations, but did not differ significantly between apoE phenotypes. The strongest relationship was found in the E3E4 group (r=0.50; P=0.001), the phenotype for which hypertriglyceridaemia is expected to be the result of increased triacylglycerol synthesis. In a stepwise multiple regression analysis, serum triacylglycerol concentrations were found to be the strongest predictor of PChE activity in the E3E4 group. In study II, PChE activity decreased as a result of bezafibrate treatment in all three apoE groups. The decrease in PChE activity with bezafibrate treatment paralleled the decrease in serum triacylglycerol concentrations in the apoE subgroups. Pravastatin treatment did not significantly affect PChE activity. Thus the present study suggests an association between PChE activity and the rate of triacylglycerol synthesis. Measurement of PChE activity may therefore be a useful tool in the choice of drug for treatment of hypertriglyceridaemia in patients with Type II diabetes. PMID- 11410112 TI - Effects of high-dose glucose-insulin-potassium on myocardial metabolism after coronary surgery in patients with Type II diabetes. AB - The effects of glucose-insulin-potassium (GIK) on cardiac metabolism have been studied previously in non-diabetic patients after cardiac surgery. Although patients with diabetes mellitus can be expected to benefit most from such treatment, the impact of GIK in diabetic patients undergoing cardiac surgery remains unexplored. Therefore the present study investigates the effects of high dose GIK on myocardial substrate utilization after coronary surgery in patients with Type II diabetes. A total of 20 patients with Type II diabetes undergoing elective coronary surgery were randomly allocated to either post-operative high dose GIK or standard post-operative care, including insulin infusion if necessary to keep blood glucose below 10 mmol/l. Myocardial substrate utilization was studied using the coronary sinus catheter technique. Haemodynamic state was assessed with the aid of Swan-Ganz catheters. High-dose GIK caused a shift towards carbohydrate utilization, with significant lactate uptake throughout the study period and significant uptake of glucose after 4 h. Arterial levels of non esterified fatty acids and beta-hydroxybutyric acid decreased, and after 1 h no significant uptake of these substrates was found. Increases in the cardiac index and stroke volume index were found in patients treated with high-dose GIK. A decrease in systemic vascular resistance was found both in the control group and in the high-dose GIK group. We conclude that high-dose GIK can be used in diabetic patients after cardiac surgery to promote carbohydrate uptake at the expense of non-esterified fatty acids and beta-hydroxybutyric acid. This could have implications for treatment of the diabetic heart in association with surgery and ischaemia. PMID- 11410113 TI - Effect of adrenomedullin on the production of endothelin-1 and on its vasoconstrictor action in resistance arteries: evidence for a receptor-specific functional interaction in patients with heart failure. AB - Endothelin-1 (ET-1) and adrenomedullin (ADM) are both produced in the arterial wall, but have opposing biological actions. Evidence from experimental animals suggests a functional interaction between ET-1 and ADM. We have tested this in humans. Small resistance arteries were obtained from gluteal biopsies taken from patients with chronic heart failure (CHF) due to coronary heart disease (CHD), or with CHD and preserved ventricular function. The contractile responses to big ET 1 and to ET-1 in both sets of vessels were studied in the absence (control) and presence of ADM at 20 pmol/l (low ADM) or 200 pmol/l (high ADM), using wire myography. ADM did not affect the conversion of big ET-1 into ET-1 in vessels from patients with either CHD or CHF. Low ADM did not alter the contractile response to ET-1 in vessels from patients with CHF. Low ADM was not tested in vessels from patients with CHD, but high ADM did not affect this response in arteries from these patients. High ADM did, however, significantly reduce the vasoconstrictor effect of ET-1 in vessels from patients with CHF. The maximum response, as a percentage of the response to high potassium, was 199% (S.E.M. 25%) in the control experiments (n=14), 205% (27%) in the low-ADM (n=7) studies and 150% (17%) in the high-ADM (n=6) experiments (P<0.001). Furthermore, the Hill coefficient increased from 0.57+/-0.05 in the absence of ADM to 1.16+/-0.15 in the high-ADM experiments, indicating that ADM at 200 pmol/l specifically antagonized one receptor type in vessels from patients with CHF. We conclude that there is a one-site receptor interaction between ADM and ET-1 that is specific for vessels from patients with CHF. This functional interaction between ADM and ET-1 in resistance arteries may be of pathophysiological importance in CHF. PMID- 11410114 TI - Lisinopril improves endothelial function in chronic cigarette smokers. AB - Cigarette smoking is a pernicious risk factor for the pathogenesis of coronary artery disease, and endothelial dysfunction is an important antecedent event in this process. This is important, as cigarette smoke is directly toxic to endothelial cells. Inhibitors of angiotensin-converting enzyme (ACE) have been shown to improve endothelial function in diabetes and hypercholesterolaemia, and are a promising option in smokers. We treated 23 subjects (age 38+/-12 years; mean+/-S.D.) for 8 weeks with 20 mg of lisinopril in a randomized controlled trial. Endothelial function was assessed by measurement of forearm blood flow responses to intra-arterial infusions of endothelial-dependent and -independent vasodilators and an endothelial-dependent vasoconstrictor [acetylcholine, sodium nitroprusside and monomethyl-L-arginine (L-NMMA) respectively] using venous occlusion plethysmography. Lisinopril significantly increased the forearm blood flow response to acetylcholine by 20% [lisinopril, 3.12+/-0.37 (mean+/-S.E.M.); placebo, 2.58+/-0.25; P=0.02, 95% confidence intervals (CI) 0.09, 1.06] (values given are ratios of flow in the infused arm to that in the control arm); there was no effect on the response to sodium nitroprusside (lisinopril, 3.97+/-0.40; placebo, 3.92+/-0.39; P=0.84; 95% CI -0.50, 0.61). The vasoconstrictor response to L-NMMA demonstrated a significant improvement (lisinopril, 0.77+/-0.06; placebo, 0.95+/-0.05; P<0.001; 95% CI -0.09, -0.27). In conclusion, these results indicate that ACE inhibition can improve endothelial function in cigarette smokers. We show that lisinopril improves both receptor-mediated and tonic NO release. The mechanism could be either that lisinopril limits the angiotensin II induced production of superoxide radicals which would normally inactivate NO, or that lisinopril may increase bradykinin-mediated NO release. PMID- 11410115 TI - Diminished skin blood flow in Type I diabetes: evidence for non-endothelium dependent dysfunction. AB - The purpose of this study was to quantify the extent to which skin blood flow (SBF) responses to application of endothelium-dependent and -independent vasodilating agents differ between Type I diabetic patients and healthy subjects. Patients and matched controls were studied after an overnight fast. SBF was determined with laser Doppler perfusion imaging before and after iontophoresis of acetylcholine (Ach; endothelium-dependent) and sodium nitroprusside (SNP; endothelium-independent). Basal SBF did not differ significantly between groups. Iontophoresis of ACh and SNP increased SBF 20-fold in controls. In the patients, the increases in SBF following iontophoresis of ACh and SNP were reduced by 18% and 19%, respectively, versus controls (P<0.05 for both). These data demonstrate that Type I diabetic patients have similar diminished SBF responses to iontophoresis of ACh and SNP, which suggests that non-endothelial-dependent factors are primarily responsible for the diminished SBF responses. PMID- 11410116 TI - Relationship of maternal protein turnover and lean body mass during pregnancy and birth length. AB - Epidemiological evidence shows that small size at birth is associated with an increased risk of developing cardiovascular and metabolic disease in adult life. We have examined the relationships between size at birth and maternal body composition and protein turnover in normal pregnant women. A group of 27 multiparous Caucasian women with singleton pregnancies were studied at around 18 and 28 weeks' gestation. Body composition was determined by anthropometry, and whole-body protein turnover was estimated by using a single oral dose of [(15)N]glycine and the end-product method. The baby's weight and length were measured within 48 h of birth. Mothers with a greater lean body mass had higher rates of protein turnover at 18 weeks' gestation. This association was largely accounted for by differences in the mother's visceral, rather than muscle, mass. Mothers who had higher protein turnover at 18 weeks' gestation had babies that were longer at birth. After adjustment for the duration of gestation and the baby's sex, 26% of the variation in length at birth was accounted for by maternal protein synthesis at 18 weeks' gestation. Maternal protein intake was not associated with the baby's birth length. Thus the mother's ability to nourish her fetus is influenced by her body composition and her rate of protein turnover. Dietary intake does not adequately characterize this ability. PMID- 11410117 TI - Effect of salt intake on endothelium-derived factors in a group of patients with essential hypertension. AB - The aim of the present study was to evaluate the effects of the level of salt intake on endothelium-derived factors in a group of patients with essential hypertension. A group of 50 patients with essential hypertension who had never been treated for the condition were placed on a low-sodium (50 mmol/day), low nitrate (400 micromol/day) diet, which was supplemented, in a single-blind fashion, with placebo tablets for the first 7 days and then with NaCl tablets (200 mmol/day) for a further 7 days (total sodium intake 250 mmol/day). At the end of both periods, 24-h ambulatory blood pressure monitoring was performed. In addition, plasma levels and 24-h urinary excretion of nitrites plus nitrates and cGMP were measured, along with plasma levels of endothelin. A high salt intake promoted significant decreases in plasma levels of nitrites plus nitrates (from 41.0+/-2.1 to 32.8+/-1.8 nmol/ml; P<0.001), 24-h urinary nitrate excretion (from 417+/-36 to 334+/-37 micromol/24 h; P=0.045) and plasma endothelin levels (from 5.6+/-0.3 to 4.6+/-0.3 pg/ml; P=0.007). The plasma concentration and 24-h urinary excretion of cGMP were not altered significantly by a high salt intake. We did not find any relationship between endothelium-derived products and 24-h mean blood pressure, at either low or high salt intakes, or between changes induced by the high-salt diet. A high salt intake also induced significant decreases in plasma renin activity, angiotensin II and aldosterone, and a significant increase in atrial natriuretic peptide. We conclude that a high salt intake decreases the plasma concentration and urinary excretion of nitrates and plasma levels of endothelin in patients with essential hypertension, suggesting that the level of salt intake may affect endothelial cell function. However, these alterations are not correlated with changes in blood pressure induced by the high salt intake. PMID- 11410118 TI - Physiological relationships between central vascular haemodynamics and left ventricular structure. AB - Left ventricular hypertrophy is an independent cardiovascular risk factor. In hypertensives, the pattern of hypertrophy is influenced by central haemodynamic characteristics. Central haemodynamics may also determine physiological differences in left ventricular structure and predispose to particular responses of the left ventricle to pathological increases in load. M-mode echocardiography was used to measure left ventricular diastolic dimension and to estimate left ventricular mass index, relative wall thickness and stroke volume in 159 healthy volunteers aged between 19 and 74 years. Tonometric sphygmography was used to estimate augmentation index, central end-systolic and mean arterial blood pressure. Effective arterial elastance was calculated as the ratio of end systolic pressure to stroke volume. Left ventricular mass index and relative wall thickness were adjusted for variation in age, sex and blood pressure before analyses. Left ventricular diastolic dimension exhibited significant inverse correlations with both effective arterial elastance (r=-0.72, P<0.0001) and augmentation index (r=-0.23, P=0.004). Adjusted left ventricular mass index was inversely correlated with effective arterial elastance (r=-0.35, P<0.0001), but no correlation was observed between left ventricular mass index and augmentation index (r=0.04). Adjusted relative wall thickness correlated with increasing effective arterial elastance (r=0.32, P<0.0001) and augmentation index (r=0.18, P=0.02). Relative wall thickness (r=0.34, P<0.0001), but not left ventricular mass index, correlated with age. Higher elastance and augmentation correlates with relatively smaller left ventricular cavity size but larger relative wall thickness. Age-related changes in left ventricular afterload may affect relative wall thickness more significantly than left ventricular mass index and may contribute to a particular change in left ventricular geometry with age. PMID- 11410119 TI - Polymorphism in the gelatinase B gene and the severity of coronary arterial stenosis. AB - Gelatinase B, as one of the matrix metalloproteinases, may be relevant to atherogenic plaque development and stability. Recently, a C-1562T substitution in the regulatory region of the gelatinase B gene was shown to up-regulate gelatinase B expression, which could be relevant to both the severity and stability of atherosclerotic plaques. We determined the genotype of 788 angiographically documented Caucasian patients with coronary artery disease (583 males and 205 females; age 56.7+/-0.4 years). The proportions of C/C (77.1%), C/T (21.4%) and T/T (1.5%) genotypes were in Hardy-Weinberg equilibrium, and did not differ between males and females (P>0.05). The frequencies of the rare T allele in patients with angiographically documented coronary artery disease (0.123), a past history of myocardial infarction (0.128) or unstable angina (0.128) were not significantly different from those in patients without such events (0.121, 0.118 and 0.128 respectively; P>0.05). In addition, the rare allele frequencies among patients with no (0.128), one (0.124), two (0.108) or three (0.121) significantly diseased vessels (> or =50% luminal obstruction) were not statistically different (P=0.932). However, the male rare T/T homozygotes had lower waist/hip ratios and levels of high-density lipoprotein cholesterol (HDL-C), and higher total cholesterol/HDL-C ratios, than C/C homozygotes (P<0.05). In conclusion, our study in a large series of angiographically defined patients suggests that the C-1562T polymorphism may not be useful as a predictor of the presence and severity of coronary atherosclerosis. PMID- 11410120 TI - Effects of combined oral hormone replacement therapy on tissue factor pathway inhibitor and factor VII. AB - Oral combined hormone replacement therapy (HRT) with oestradiol and norethisterone increases plasma levels of prothrombin fragment 1+2 (F1+2), indicating an increase in thrombin generation, but the mechanisms underlying this increase are uncertain. The aim of this randomized, placebo-controlled study was to determine whether an increase in factor VII, a factor that combines with tissue factor to activate the extrinsic pathway, or a decrease in tissue factor pathway inhibitor (TFPI), an inhibitor of extrinsic pathway activation, may contribute to increases in thrombin generation occurring with HRT. Healthy postmenopausal women aged 50-75 years received placebo (n=19) or oral combined HRT (n=18) and had blood collected for measurement of factor VII coagulation activity (VIIc), activated factor VII (VIIa) and TFPI at baseline and at 6 weeks. Baseline characteristics were similar in the two groups, including age, body mass index and cholesterol levels. As reported previously, HRT increased the F1+2 concentration by 20%. Placebo had no effect on VIIc, VIIa or TFPI, but 6 weeks of combined HRT decreased VIIc [from 1.11+/-0.06 (mean+/-S.E.M.) to 1.03+/-0.06 i.u./ml; P<0.03], VIIa [from 43.9; 10.8-198.3 (median; range) to 35.0; 6.3-66.8 m units/ml; P<0.03] and TFPI [from 81.3+/-6.5 to 60.4+/-5.5 ng/ml; P<0.0001]. The decrease in TPFI with HRT was not correlated with the elevation in F1+2 levels. In conclusion, the increase in thrombin generation seen with HRT is not due to an effect on factor VII; in addition, while a contribution from the decrease in TFPI is possible, increased thrombin generation is not directly related to the decrease in TFPI. PMID- 11410121 TI - Effect of salt intake on endothelium-derived factors in a group of essential hypertensive patients. PMID- 11410122 TI - Differing thresholds for modulatory effects of adrenomedullin infusion on haemodynamic and hormone responses to angiotensin II and adrenocorticotrophic hormone in healthy volunteers. AB - Experimental data indicate that adrenomedullin (AM) interacts at various levels with the renin-angiotensin-aldosterone system and the hypothalamic-pituitary adrenal axis, but data from humans are scant. We examined the effects of intermediate-dose, short-term AM infusion on angiotensin II- and adrenocorticotrophic hormone (ACTH)-mediated hormone and haemodynamic responses in healthy subjects. Seven normal volunteers (age 18-25 years) completed a placebo-controlled crossover study. Each subject was studied on day 4 of two periods of a low-salt diet (40 mmol of sodium and 80 mmol of potassium daily), receiving incremental infusions of angiotensin II in the morning and ACTH in the afternoon of each study day, on a background infusion of AM (4 pmol.min(-1).kg( 1)) or vehicle (hemaccel). Achieved plasma AM levels (23+/-6 pmol/l) and peak angiotensin II levels (160 pmol/l) were similar on the two experimental days. While the pressor action of angiotensin II was attenuated by AM (P<0.01) and noradrenaline levels rose (P<0.05), the aldosterone response was unaltered. During ACTH infusion, AM increased heart rate (P<0.01), plasma adrenaline (P<0.01) and plasma noradrenaline (P<0.05), and augmented the cortisol response (P<0.01), but was without effect on aldosterone levels and blood pressure. We conclude that the threshold for the effects of AM on aldosterone secretion in humans is set higher than for other biological responses to this hormone, namely blood pressure, heart rate, sympathetic activity and cortisol secretion, under these experimental conditions. PMID- 11410123 TI - A conditional form of Bruton's tyrosine kinase is sufficient to activate multiple downstream signaling pathways via PLC Gamma 2 in B cells. AB - BACKGROUND: Bruton's tyrosine kinase (Btk) is essential for B cell development and function. Mutations of Btk elicit X-linked agammaglobulinemia in humans and X linked immunodeficiency in the mouse. Btk has been proposed to participate in B cell antigen receptor-induced signaling events leading to activation of phospholipase C-gamma2 (PLCgamma2) and calcium mobilization. However it is unclear whether Btk activation is alone sufficient for these signaling events, and whether Btk can activate additional pathways that do not involve PLCgamma2. To address such issues we have generated Btk:ER, a conditionally active form of the kinase, and expressed it in the PLCgamma2-deficient DT40 B cell line. RESULTS: Activation of Btk:ER was sufficient to induce multiple B cell signaling pathways in PLCgamma2-sufficient DT40 cells. These included tyrosine phosphorylation of PLCgamma2, mobilization of intracellular calcium, activation of extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK) mitogen-activated protein kinase (MAPK) pathways, and apoptosis. In DT40 B cells deficient for PLCgamma2, Btk:ER activation failed to induce the signaling events described above with the consequence that the cells failed to undergo apoptosis. CONCLUSIONS: These data suggest that Btk:ER regulates downstream signaling pathways primarily via PLCgamma2 in B cells. While it is not known whether activated Btk:ER precisely mimics activated Btk, this conditional system will likely facilitate the dissection of the role of Btk and its family members in a variety of biological processes in many different cell types. PMID- 11410124 TI - In vitro study of a multi-layer piezoelectric crystal attic hearing implant. AB - We have developed a prototype middle-ear hearing implant which uses a multilayer piezoelectric actuator. In this series of experiments the actuator was attached to the medial wall of the attic so that it makes contact with the body of the incus. Initial in vitro evaluation has been carried out using a laser vibrometer (Polytec CLV) to measure stapes velocity. Stapes displacement is calculated by mathematical integration. The device used in this way is particularly effective at transmitting high frequency sound to the stapes. When switched off the actuator impairs the transmission of sound to the ossicular chain at low frequencies, but this effect is only 7 dB at most. The stapes displacements resulting from the action of the implant have a linear relationship with the voltages used to drive the system. The high capacitance of the present actuator means that its power requirements are higher than that of other comparable devices. An optimal method of coupling the device to the incus has yet to be identified. PMID- 11410125 TI - Nebulized surfactant as a treatment choice for otitis media with effusion: an experimental study in the rabbit. AB - Exogenous surfactant can improve eustachian tube function in experimentally induced otitis media with effusion (OME). Performing tympanometric recordings, the efficacy of inhaled nebulized surfactant, as compared with inhaled nebulized physiological saline was investigated, for the treatment of OME experimentally induced in the rabbit by intrabullar inoculation of heat-killed Streptococcus pneumoniae. In addition, the histological changes in middle ears after the treatment were investigated in order to establish whether the pathological findings correlated with the results. Middle-ear pressure values before, and after, treatment were analyzed by the Wilcoxon statistical method, and the Mann Whitney U test was used to compare the post-treatment values between groups. In all ears with OME in the affected animals, which were treated with nebulized surfactant inhalation, a positively significant (p<0.05) increase of pressure more than 20 daPa was recorded. In the control group, after inhalation of nebulized physiological saline, there was no positive increase in the affected middle-ear pressures; on the contrary, more negative pressure changes were recorded. In the histological evaluation, middle-ear epithelia and sub-epithelial space were normal in surfactant-treated ears with OME, whereas mucosal thickening with an oedematous sub-epithelial space containing occasional inflammatory cells and increases in connective tissue and vascularity, and effusions on the epithelial surface were present in the ears with OME in the control group. The significant improvement in the negative middle-ear pressure after nebulized surfactant treatment and the histological findings shown in our study can support the theory that surface-active agents are of importance in eustachian tube function even under pathologic conditions, such as OME. PMID- 11410126 TI - Myringoplasty in children: anatomical and functional results. AB - To assess the results of myringoplasty in children and determine the factors influencing post-operative results a retrospective study of the anatomical and functional results of 41 myringoplasties in children was performed, considering only the cases of uncomplicated perforation that did not require ossiculoplasty or mastoidectomy. The overall success rate was 80.5 per cent after a mean follow up of 39 months. The mean post-operative air conduction threshold significantly improved in the successful cases with a mean audiological improvement of 11 dB (p<0.05). No post-operative sensorineural hearing loss was observed. There was a significant statistical association between the presence of a dry ear at the time of surgery and good surgical results (p<0.01). Surgical outcome was not affected by the patient's age, the site and size of the perforation, previous adenoidectomy, surgical technique (overlay vs underlay), or the status of the contralateral ear. Our findings suggest that myringoplasty is a valid procedure in the paediatric population that gives good anatomical and functional results. The status of the middle ear (i.e. the presence of a dry ear), significantly improves surgical outcome; and so careful inflammatory changes in the middle-ear mucosa should be evaluated and medical treatment considered before surgery. PMID- 11410127 TI - Temporal bone dissection: a possible route for prion transmission? AB - The aim of this study was to determine whether neural tissue is present in the bone 'dust' given off during temporal bone drilling. Bone 'dust' from three temporal bone dissections was collected and examined. Evidence of neural tissue was present in two out of the three specimens. Neural tissue is present in the bone dust given off during temporal bone drilling. This poses the question as to the risk of prion transmission during such dissection. PMID- 11410128 TI - Nerve origin of the acoustic neuroma. AB - Two hundred and seventy-one Japanese patients who underwent surgical removal of neuroma from the cerebello-pontine angle using the translabyrinthine approach were retrospectively reviewed in order to investigate the nerve origin on an anatomical basis. In 269 out of the 271 cases, the origin of the neuromas was identified. Among these cases, 84.8 per cent of tumours originated from the inferior vestibular nerve (IVN), followed by the superior vestibular nerve (SVN) (8.9 per cent). Of the 5.6 per cent of tumours arising from the vestibular nerve, however, it could not be determined whether they originated from the SVN or IVN. Two cases of neuromas originating from the facial nerve were seen (0.7 per cent). No neuroma arose from the cochlear nerve. Currently, the diagnosis of acoustic neuromas is best made with Gd-enhanced magnetic resonance imaging (MRI). However, our data indicate that the development of a functional test of the individual IVN can be useful for screening most cases of acoustic neuroma and in facilitating their early diagnosis. PMID- 11410129 TI - Endoscopy in the assessment of children with nasal obstruction. AB - Nasal obstruction is common in children, and is often attributed to adenoid enlargement. This prospective study was performed to determine whether routine nasal endoscopy is of value for children undergoing surgery for nasal obstruction. Forty-eight children aged two to nine years undergoing adenoidectomy, and six normal controls, were examined under general anaesthesia with a 4 mm rigid endoscope. A video of the endoscopy was subsequently assessed independently by an observer blinded to the original findings and the presence of nasal symptoms. The endoscopist and independent assessor were in agreement regarding 86 per cent of the findings. Three quarters of the children had abnormalities on endoscopy in addition to enlarged adenoids, and in 23 per cent these were potentially of major clinical significance (unsuspected foreign body, gross septal deviation, gross hypertrophy of the turbinates). Endoscopy produced no post-operative complications and was possible in children as young as two years of age, without decongestants. Nasal endoscopy is a safe, objective and useful means of identifying potentially significant abnormalities in children with nasal obstruction. PMID- 11410130 TI - Endoscopic transnasal neurolytic sphenopalatine ganglion block for head and neck cancer pain. AB - Nasal endoscopy is a valuable adjunct to the localization of the sphenopalatine ganglion. Twenty-two patients with advanced malignancies of the head and neck region whose pain was not adequately controlled with conventional medications, including oral morphine, were given nasal endoscopically guided neurolytic sphenopalatine ganglion block with six per cent phenol after a prognostic block with local anaesthetic solution. Seventeen patients had good immediate relief. One had partial relief and four had inadequate relief. On follow-up for one month, the patients had significantly lower pain intensity and the pain was more manageable with oral medication. The vicious cycle of pain was broken. This block is relatively safe and can be usefully performed as an out-patient procedure. PMID- 11410131 TI - Supracricoid partial laryngectomy for non-squamous cell carcinoma of the larynx. AB - The objective of this paper was to evaluate the potential utility of supracricoid partial laryngectomies (SCPLS) for non-squamous cell carcinoma of the larynx. To illustrate our management of such tumours we present a case series based on six patients. Local control was achieved in five patients, with the sixth being salvaged by total laryngectomy and post-operative radiation therapy. Three of the six patients died of distant metastases. We concluded that supracricoid partial laryngectomies should become part of the armamentarium of the otolaryngologist - head and neck surgeon for non-squamous cell carcinoma of the larynx. PMID- 11410132 TI - Reasons for success or failure in surgical voice restoration after total laryngectomy: an Australian study. AB - Over the past 20 years, use of tracheo-oesophageal puncture (TEP) speech after total laryngectomy has resulted in reported success rates of 90-93 per cent worldwide. Despite this, data collected from major acute hospitals in Victoria, Australia indicated that, of 38 patients who underwent total laryngectomy in 1997, only 10 (26 per cent) were using TEP speech as their primary mode of communication at 12 months post-operatively. This paper describes how a quantitative research methodology was used to investigate why so few patients in Victoria were successfully using TEP speech as their chosen mode of rehabilitation after total laryngectomy. Patients, speech pathologists and ENT surgeons were interviewed. Their thoughts and beliefs regarding speech rehabilitation were mapped, and themes were identified, coded and analysed. This paper describes and discusses the results of this research and its possible implications for future patient management, through establishing a model for 'ideal' speech rehabilitation. PMID- 11410133 TI - Cyclin-dependent kinase inhibitor p27 is related to cell proliferation and prognosis in laryngeal squamous cell carcinomas. AB - An immunohistochemical study of both p27 and proliferating cell nuclear antigen (PCNA) was performed on 102 cases of laryngeal squamous cell carcinomas (LSCCs), and large variations in p27 expression were found among the tumours. Reduced expression of p27 was revealed in 54 (52.9 per cent) cases and correlated with tumour size, lymph node metastasis, and clinical stage, but did not correlate with age, sex, tumour site, or tumour grade. A significant positive correlation was found between the percentage of loss of p27 expression and the PCNA index (r = 0.844, p < 0.0001). Reduced expression of p27 was significantly correlated with both reducing disease-free and overall survival by univariate analysis. By multivariate analysis, reduced expression of p27, tumour grade, tumour size, lymph node metastasis as well as clinical stage were independent prognostic factors for overall survival of LSCCs. These findings indicate that reduced expression of p27 may be correlated with the malignant biological behaviour of LSCCs. PMID- 11410134 TI - Raising of the mucoperichondrial flap in septal surgery. PMID- 11410135 TI - Safe and easy method for secondary tracheo-oesophageal puncture. AB - We describe a method for performing secondary tracheo-oesophageal puncture in patients who have undergone total laryngectomy. The technique is straightforward and easy. It uses a minimum of readily available equipment with a simple modification to a standard pharyngoscope. PMID- 11410136 TI - Severe audiovestibular loss following ear syringing for wax removal. AB - Syringing of the ear is one of the commonest procedures performed for cleaning cerumen from the external auditory canal. Common complications following syringing are pain, external auditory canal trauma and otitis externa. Hearing and vestibular loss are often mentioned as a complication in descriptions of this technique, but we have not been able to find a reported case of such an occurrence. We report one such a case. PMID- 11410137 TI - Spontaneous recovery of profound post-meningitic hearing loss. AB - The timing of insertion of a cochlear implant (CI) in post-meningitic sensorineural hearing loss is, in spite of recent advances in the understanding of the pathogenesis of the condition, still controversial. The danger of labyrinthitis ossificans allows only a little time to decide whether to implant a CI or not. On the other hand the clinician needs to be certain that no residual hearing is present and whether or not the patient will benefit from conventional amplification. A well-documented case of early spontaneous recovery of a profound post-meningitic sensorineural hearing loss in a child is presented and the relevant literature reviewed. PMID- 11410138 TI - Spontaneous temporal encephaloceles masked by dual pathology: report of two cases. AB - Spontaneous temporal meningoencephaloceles are rare entities and diagnostic difficulties can occur. We present two cases whose presentation was atypical and diagnosis delayed by the presence of dual pathology. PMID- 11410139 TI - Osseous haemangioma of inferior turbinate. AB - We present a case of osseous haemangioma arising within the inferior turbinate. PMID- 11410140 TI - Solitary infantile myofibromatosis in the lateral orbit floor showing spontaneous regression. AB - Infantile myofibromatosis is a rare benign tumour usually occurring early in infancy. We describe the case of a 10-year-old boy with solitary infantile myofibromatosis in the left lateral orbit floor which regressed spontaneously. Although our patient was older than previously reported cases and showed bony destruction confirmed by computed tomography (CT), this tumour was diagnosed as infantile myofibromatosis based on immunohistochemical findings. The tumour disappeared spontaneously six months after incisional biopsy, that also indicated this tumour was an infantile myofibromatosis. PMID- 11410141 TI - Cervical osteophytes presenting as unilateral vocal fold paralysis and dysphagia. AB - Any process involving either the vagus nerve, its recurrent laryngeal branch or the external branch of the superior laryngeal nerve may cause paralysis of the vocal fold. The most common cause is neoplasm. Clinically, the patients often present with a hoarse, breathy voice as well as symptoms of aspiration. The following represents a unique case of unilateral vocal fold paralysis and dysphagia caused by a degenerative disease of the cervical spine, resluting in extrinsic compression of the recurrent laryngeal nerve. PMID- 11410142 TI - Persistent foreign body sensation and pharyngeal pain due to retention of staples: an interesting sequelae of endoscopic stapling procedure. AB - The surgical treatment of a pharyngeal pouch with endoscopic stapling diverticulotomy is a relatively new concept. Long-term results and complications are yet to be fully studied. We describe a patient who developed persistent pharyngeal pain and foreign body sensation due to retention of a clump of staples at the cricopharyngeal sphincter. This complication has not been reported before. This case highlights the need for repeat endoscopy rather than a barium swallow X ray if the patients are symptomatic after stapling procedures. PMID- 11410143 TI - Actinomycosis of the post-cricoid space: an unusual cause of dysphagia. AB - Cervicofacial actinomycosis is known to affect many soft tissues and bony structures in the head and neck. However to the authors' knowledge, actinomycosis of the post-cricoid region has not been previously reported. A case of a 74-year old male who developed actinomycosis of the post-cricoid region after radiotherapy for a laryngeal carcinoma is presented. Actinomycosis should be considered in the differential diagnosis of dysphagia following radiotherapy for squamous cell carcinoma of the larynx, as early treatment is likely to result in a favourable outcome. PMID- 11410144 TI - Combined 'trache-stent': a useful option in the treatment of a complex case of subglottic stenosis. AB - The authors describe the adaptation of a Montgomery T-tube laryngo-tracheal stent to incorporate an uncuffed fenestrated Shiley tracheostomy tube in the management of a complex case of subglottic stenosis. The combined 'trache-stent' provided a secure, unobstructed airway and optimal phonation. The presence of a removable inner tube facilitated cleaning and reduced patient anxiety regarding the perceived risks of stent obstruction with dried secretions. The combined 'trache stent' was upsized at four weeks with minimal evidence of local granulation formation. The device was removed entirely six weeks later after direct laryngoscopy reconfirmed the above findings. Almost two years later the patient retains a good voice and airway using a simple fenestrated tracheostomy tube and speaking valve. The subglottic area is stable and the patient has returned to full-time employment. PMID- 11410145 TI - Epithelial-myoepithelial carcinoma of the parotid gland: a case report and review of the cytological and histological features. AB - Epithelial-myoepithelial carcinoma (EMC) is a rare biphasic tumour of the salivary glands typically arising in the parotid. Fine needle aspiration cytology is widely used in the initial investigation of salivary gland swellings and whilst the cytological features of this tumour have been described they are not well recognized. This report describes the clinicopathological features of a case of epithelial-myoepithelial carcinoma of the parotid gland and highlights the importance of awareness of this tumour in the differential diagnosis of biphasic tumours on fine needle aspiration cytology. PMID- 11410146 TI - Cytotoxic T-cell abundance and virus load in human immunodeficiency virus type 1 and human T-cell leukaemia virus type 1. AB - The correlation between virus load and specific cytotoxic T-lymphocyte (CTL) frequency during the chronic phase in human immunodeficiency virus type 1 (HIV-1) infection has been found to be negative in cross-sectional studies. We report here that, in infection with the related retrovirus human T-cell leukaemia virus type 1 (HTLV-1), the correlation is positive in asymptomatic carriers and zero in patients with the associated inflammatory disease HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). We demonstrate that the direction of the correlation may depend on the efficacy of the CTL response using mathematical models. We conclude that the CTL response is effective in asymptomatic carriers of HTLV-1, but ineffective in patients with HAM/TSP. Virus mediated impairment of specific CTL production in HIV-1 infection can account for the negative correlation observed. PMID- 11410147 TI - Multiple dynamics in a single predator-prey system: experimental effects of food quality. AB - Recent work with the freshwater zooplankton Daphnia has suggested that the quality of its algal prey can have a significant effect on its demographic rates and life-history patterns. Predator-prey theory linking food quantity and food quality predicts that a single system should be able to display two distinct patterns of population dynamics. One pattern is predicted to have high herbivore and low algal biomass dynamics (high HBD), whereas the other is predicted to have low herbivore and high algal biomass dynamics (low HBD). Despite these predictions and the stoichiometric evidence that many phytoplankton communities may have poor access to food of quality, there have been few tests of whether a dynamic predator-prey system can display both of these distinct patterns. Here we report, to the authors' knowledge, the first evidence for two dynamical patterns, as predicted by theory, in a single predator-prey system. We show that the high HBD is a result of food quantity effects and that the low HBD is a result of food quality effects, which are maintained by phosphorus limitation in the predator. These results provide an important link between the known effects of nutrient limitation in herbivores and the significance of prey quality in predator-prey population dynamics in natural zooplankton communities. PMID- 11410148 TI - Lifetime allocation of juvenile and adult nutritional resources to egg production in a holometabolous insect. AB - In holometabolous insects reproductive success is strongly determined by the nutritional resources available to the females. In addition to nutrients derived from adult feeding, resources for egg production may come from the limited reserves accumulated during the larval stages. The pattern of allocation of these larval reserves to egg production is expected to be strongly linked to the nutritional ecology of the adult. We investigate the temporal pattern of allocation of larval reserves to reproduction in a host-feeding parasitoid wasp. As predicted by the dynamics of allocation of an adult meal, larval reserves are the main source of nutrients for four or five days after emergence. However, despite the high frequency of host feeding and the high nutrient content of a haemolymph meal, which we predicted would lead to larval reserves being conserved in the event of host deprivation, larval reserves contribute to egg production throughout the lifetime of the female. We propose several mechanistic and adaptive explanations for our results, including the possible existence of a limiting or key nutrient for egg production of exclusively larval origin. We make further predictions concerning the pattern of allocation of larval resources in parasitoids with different adult nutritional requirements. PMID- 11410149 TI - The effect of transient food stress on female mate preference in the stalk-eyed fly Cyrtodiopsis dalmanni. AB - The effect of transient nutritional stress (sucrose culture medium) on female mate preference was investigated in the stalk-eyed fly Cyrtodiopsis dalmanni. Two experimental treatments were run in parallel. In the first, female preference was initially tested on corn (standard medium) and subsequently on sucrose (novel medium). In the second, female preference was tested in the reverse order: sucrose then corn. Females fed on corn had stronger preferences for large-eyespan males than females fed on sucrose. Female age, experience or the order of exposure to culture media had no effects on the strength of preference. An additional experiment demonstrated that the sucrose diet reduced the number of developing and mature eggs. We discuss the adaptive significance of changes in female preference due to transient nutritional stress, and the effect of changes in female preference on the strength of sexual selection. PMID- 11410150 TI - Carotenoid scarcity, synthetic pteridine pigments and the evolution of sexual coloration in guppies (Poecilia reticulata). AB - Carotenoid-based sexual coloration is the classic example of an honest signal of mate quality. Animals cannot synthesize carotenoid pigments and ultimately depend on dietary sources. Thus, in carotenoid-poor environments, carotenoid coloration may be a direct indicator of foraging ability and an indirect indicator of health and vigour. Carotenoid coloration may also be affected, more directly, by parasites in some species. Carotenoids are not, however, the only conspicuous pigments available to animals. Pteridine pigments, with similar spectral properties, are displayed in the exoskeletons and wings of insects, the irides of birds and the skins of fishes, lizards and amphibians. Unlike carotenoids, pteridines are synthesized de novo by animals. We report that the orange spots that male guppies (Poecilia reticulata) display to females contain red pteridine pigments (drosopterins) in addition to carotenoids. We also examined the relationship between drosopterin production by males and carotenoid availability in the field. The results contrasted sharply with the hypothesis that males use drosopterins to compensate for carotenoid scarcity: males used more, not less, drosopterins in streams with higher carotenoid availability. The positive association between drosopterin use and carotenoid availability could reflect the costs of drosopterin synthesis or it could be a consequence of females preferring a particular pigment ratio or hue. Male guppies appear to use drosopterin pigments in a manner that dilutes, but does not eliminate, the indicator value of carotenoid coloration. PMID- 11410151 TI - Male mating preference for female survivorship in the seaweed fly Gluma musgravei (Diptera: Coelopidae). AB - The seaweed fly mating system is characterized by pre-mating struggles during which females exhibit a mate rejection response involving kicking, shaking and abdominal curling. Males must resist rejection until females become passive and allow copulation to take place. However, despite the vigorous nature of the struggle males frequently dismount passive females without attempting copulation. Here we show that rejected females suffered higher post-encounter mortality rates than those accepted by males in the seaweed fly Gluma musgravei. Furthermore, we show that males also preferentially mounted females with higher future longevity. We propose that this male mate choice for female survivorship has evolved as a result of females often having to survive for long periods after mating until suitable oviposition sites become available. Such male preferences for female survivorship may be common in species in which oviposition must sometimes be substantially delayed after mating. PMID- 11410152 TI - Reinforcement during ecological speciation. AB - Reinforcement of pre-zygotic isolation can result when any of several kinds of selection act against hybrids. This paper investigates the situation where hybrids are selected against for ecological reasons, for example when there is no niche for individuals that are phenotypically intermediate between the parental species. The calculations here show how much ecological selection can lead to the reinforcement of a female mating preference or an assortative mating trait that is expressed in both sexes. The model allows for the ecological trait to be affected by any number of loci, but assumes that selection is weak and the introgression rate small. The effect of selection against hybrids increases rapidly as the difference between the mean phenotypes of the two populations increases. When genetic variation in the ecological trait is caused by many loci, stabilizing selection on it further contributes to reinforcement. PMID- 11410153 TI - Ejaculate allocation by male sand martins, Riparia riparia. AB - Males of many species allocate sperm to ejaculates strategically in response to variation in the risk and intensity of sperm competition. The notable exception is passerine birds, in which evidence for strategic allocation is absent. Here we report the results of a study testing for strategic ejaculate allocation in a passerine bird, the sand martin (Riparia riparia). Natural ejaculates were collected from males copulating with a model female. Ejaculates transferred in the presence of a rival male contained significantly more sperm than ejaculates transferred in the absence of a rival male. There was no evidence that this difference was due to the confounding effects of the year of ejaculate collection, the identity of the model female, the colony, the stage of season or the period of the day in which ejaculates were collected. A more detailed examination of the ejaculate patterns of individual males, achieved by the DNA profiling of ejaculates, provided additional evidence for strategic allocation of sperm. PMID- 11410154 TI - The costs of egg production and incubation in great tits (Parus major). AB - The costs of egg production and incubation may have a crucial effect on avian reproductive decisions, such as clutch size and the timing of reproduction. We carried out a brood-size enlargement experiment on the great tit (Parus major), in which the birds had to lay and incubate extra eggs (full costs), only incubate extra eggs (free eggs) or did not pay any extra cost (free chicks) in obtaining a larger brood. We used female fitness (half the recruits produced plus female survival) as a fitness measure because it is the female which pays the costs of egg production and incubation, and because clutch size is under female control. Female fitness decreased with increasing costs (fitness of free chicks females is higher than that of free eggs females which is higher than that of full costs females). These fitness differences were due to differences in female survival rather than in the number of recruits produced. This is the first time that the costs of egg production and incubation have been estimated using such a complete fitness measure, including, as our measure does, the local survival to the following year of both the female and her offspring. Our results emphasize that reproductive decisions cannot be understood without taking egg production and incubation costs into account. PMID- 11410155 TI - 'Good genes as heterozygosity': the major histocompatibility complex and mate choice in Atlantic salmon (Salmo salar). AB - According to the theory of mate choice based on heterozygosity, mates should choose each other in order to increase the heterozygosity of their offspring. In this study, we tested the 'good genes as heterozygosity' hypothesis of mate choice by documenting the mating patterns of wild Atlantic salmon (Salmo salar) using both major histocompatibility complex (MHC) and microsatellite loci. Specifically, we tested the null hypotheses that mate choice in Atlantic salmon is not dependent on the relatedness between potential partners or on the MHC similarity between mates. Three parameters were assessed: (i) the number of shared alleles between partners (x and y) at the MHC (M(xy)), (ii) the MHC amino acid genotypic distance between mates' genotypes (AA(xy)), and (iii) genetic relatedness between mates (r(xy)). We found that Atlantic salmon choose their mates in order to increase the heterozygosity of their offspring at the MHC and, more specifically, at the peptide-binding region, presumably in order to provide them with better defence against parasites and pathogens. This was supported by a significant difference between the observed and expected AA(xy) (p = 0.0486). Furthermore, mate choice was not a mechanism of overall inbreeding avoidance as genetic relatedness supported a random mating scheme (p = 0.445). This study provides the first evidence that MHC genes influence mate choice in fish. PMID- 11410156 TI - Microsatellite diversity predicts recruitment of sibling great reed warblers. AB - Inbreeding increases the level of homozygosity, which in turn might depress fitness. In addition, individuals having the same inbreeding coefficient (e.g. siblings) vary in homozygosity. The potential fitness effects of variation in homozygosity that is unrelated to the inbreeding coefficient have seldom been examined. Here, we present evidence from wild birds that genetic variation at five microsatellite loci predicts the recruitment success of siblings. Dyads of full-sibling great reed warblers (Acrocephalus arundinaceus), one individual of which became a recruit to the natal population while the other did not return, were selected for the analysis. Each dyad was matched for sex and size. Local recruitment is strongly tied to fitness in great reed warblers as the majority of offspring die before adulthood, philopatry predominates among surviving individuals and emigrants have lower lifetime fitness. Paired tests showed that recruited individuals had higher individual heterozygosity and higher genetic diversity, which was measured as the mean squared distance between microsatellite alleles (mean d(2)), than their non-recruited siblings. These relationships suggest that the microsatellite markers, which are generally assumed to be neutral, cosegregated with genes exhibiting genetic variation for fitness. PMID- 11410157 TI - The benefits of genetic diversity outweigh those of kin association in a territorial animal. AB - The theories of kin selection and heterogeneous advantage have been central to studies of altruistic behaviour and the evolution of sex over the last 35 years. Yet they predict diametrically opposite effects of genetic diversity on population density. Close relatives gain inclusive fitness advantages by preferentially associating with and behaving altruistically towards one another. However, heterogeneous advantage, which predicts competition to be highest when genetic diversity is low, suggests that benefits will be greater for individuals in groups of non-kin. Here we test how these two processes balance and affect the productivity of populations of animals in natural habitats. We report from a study of juvenile Atlantic salmon in the wild that heterogeneous advantage outweighs the benefits of kin-biased behaviour, resulting in a 1.8-fold higher population biomass and significantly better condition of individual fish. PMID- 11410158 TI - Evolutionary history and speciation modes in the cyprinid genus Barbus. AB - Phylogenetic relationships and evolutionary patterns in the genus Barbus were examined through the analysis of the complete sequences of three mitochondrial genes: ATPases 8 and 6, which overlap slightly, and cytochrome b. This complex genus includes diploid as well as tetraploid and hexaploid species that are distributed throughout the Palaearctic, Ethiopian and Asiatic biogeographical regions. Given that genome duplication is an important evolutionary mechanism in eukaryotes, in the present report we attempt to describe its role in the evolution of the genus Barbus, as well as drawing systematic and phylogenetic conclusions. The phylogenetic results indicated the splitting of the current Barbus genus into five main mitochondrial lineages corresponding to (i) the genus Barbus sensu stricto (tetraploid, which is subdivided into the subgenera Barbus and Luciobarbus), (ii) the hexaploid species, (iii) the Ethiopian tetraploid species, (iv) the African diploid species, and (v) the Asian diploid species. The branching of 'foreign' genera as sister groups of some of these monophyletic assemblages (such as Aulopyge is to Barbus sensu stricto or Varicorhinus is to the hexaploid barbels) demonstrates the polyphyly of the group. Moreover, the relationships between the proposed lineages also show that genome duplication may be considered as a homoplasic character since it must have occurred over at least three independent periods and/or in three independent areas. In relation to the possible saltational evolutionary model for the polyploid species examined here, it was found that, although feasible at the nuclear level, the mitochondrial markers looked at do not appear to have undergone this type of evolution. Rather, they seem to have experienced more or less constant change through time. PMID- 11410159 TI - Stochastic noise interferes coherently with a model biological clock and produces specific dynamic behaviour. AB - The influence of noise is unavoidable in all living systems. Its impact on a model of a biological clock, normally running in regular oscillating modes, is examined. It is shown that in a specific system in which endogenous rhythmicity is produced by a beat oscillator acting on a feedback coupled metabolic pool system, noise can act coherently to produce unexpected dynamic behaviour, running from regular over pseudo-regular to irregular time-structures. If the biological system consists of a set of identical weakly coupled cells, stochasticity may lead to phase decoupling producing irregular spatio-temporal patterns. Synchronization via phase resetting can be achieved by external short-time temperature pulses. Explicit results are obtained for the well-studied circadian photosynthesis oscillations in plants performing crassulacean acid metabolism. Because of the generic structure of the underlying nonlinear dynamics they can, however, be regarded as a general property of the influence of noise on nonlinear excitable systems with fixed points occuring close to limit cycles. PMID- 11410160 TI - Feedback control of the rate of peat formation. AB - The role of peatlands in the global carbon cycle is confounded by two inconsistencies. First, peatlands have been a large reservoir for carbon sequestered in the past, but may be either net sources or net sinks at present. Second, long-term rates of peat accumulation (and hence carbon sequestration) are surprisingly steady, despite great variability in the short-term rates of peat formation. Here, we present a feedback mechanism that can explain how fine-scale and short-term variability in peat-forming processes is constrained to give steady rates of peat accumulation over longer time-scales. The feedback mechanism depends on a humpbacked relationship between the rate of peat formation and the thickness of the aerobic surface layer (the acrotelm), such that individual microforms (hummocks, lawns, hollows and pools) expand or contract vertically in response to fluctuations in the position of the water table. Hummocks (but not hollows) 'evolve' to a steady state where changes in acrotelm thickness compensate for climate-mediated variations in surface wetness. With long-term growth of a topographically confined peat deposit, the steady state gradually shifts to a thicker acrotelm (i.e. taller hummocks) and lower rates of peat formation and carbon sequestration. PMID- 11410161 TI - Primary structure and developmental expression of zebrafish sodium channel Na(v)1.6 during neurogenesis. AB - A zebrafish sodium channel cDNA encoding a 1949-amino acid polypeptide, Na(v)1.6, was isolated. Two transcripts were detected in zebrafish adult brain but not in cardiac or skeletal muscle. The RNase protection analysis confirmed the neural specificity of zebrafish Na(v)1.6 24 hours postfertilization (hpf) Na(v)1.6 was expressed in the trigeminal ganglion, anterior and posterior lateral line ganglia, rhombomeres, and Rohon-Beard neurons. This preferential localization suggests that Na(v)1.6 plays an important role in tactile sensitivity. The abundance of zebrafish Na(v) 1.6 mRNA in the central and peripheral nervous systems increased markedly between 48 and 72 hpf, during the maturation of the nervous system. PMID- 11410162 TI - HMGN4, a newly discovered nucleosome-binding protein encoded by an intronless gene. AB - We describe a newly discovered nuclear protein, HMGN4, that is closely related to the canonical HMGN2 nucleosome-binding protein. The protein is encoded by an intronless gene, which, in humans, is located in the hereditary hemochromatosis [correction of hemachromatosis] region at position 6p21.3. A single approximately 2-kb HMGN4 mRNA was found to be expressed, in variable amounts, in all human tissues tested; however, the HMGN4 transcript was significantly less abundant than that of HMGN2. The HMGN4 protein could be detected in HeLa cells by Western analysis with an antibody elicited against a unique region of the protein. Transfection of HeLa cells with a plasmid expressing HMGN4-GFP indicated that the protein localizes to the nucleus. Our results expand the multiplicity of the HMGN protein family and increase the known cellular repertoire of nucleosome-binding proteins. PMID- 11410163 TI - Regulation of MUC1 expression in human mammary cell lines by the c-ErbB2 and ras signaling pathways. AB - The MUC1 protein is a highly O-glycosylated transmembrane molecule that is expressed at the luminal surface of most glandular epithelial cells and is upregulated in carcinomas. Here, we report the effect of the activation of the c ErbB2 --> Ras pathway on the expression of the MUC1 gene in the nontumorigenic mammary cell lines MTSV1-7 and HB2 and in the malignant cell lines T47D and ZR75. Endogenous levels of MUC1 mRNA and protein in HB2 clones permanently overexpressing c-ErbB2 or V12-H-Ras were markedly reduced compared with levels in the parental cell lines. Furthermore, in transient transfection assays, the transcription of a CAT reporter construct driven by the MUC1 promoter was inhibited when cotransfected with a c-ErbB2 or a V12-H-ras expressing vector. Transient transfections using mutant forms of the ras oncogene, and the inhibitor chemical wortmannin, indicated that the pathway activated by c-ErbB2 proceeds via activation of Ras and that the Raf and phosphoinositide 3-kinase pathways are involved. Finally, cotransfection assays using a reporter gene driven by the MUC1 promoter carrying abolishing mutations in some of the cis-acting elements showed that a GC box at -99/-91 is crucial for responsiveness to c-ErbB2 inhibition of transcription. PMID- 11410164 TI - Molecular cloning and preliminary functional analysis of two novel human KRAB zinc finger proteins, HKr18 and HKr19. AB - cDNA clones encoding two novel human KRAB zinc finger proteins, HKr18 and HKr19, were isolated from a human testis cDNA library. Their corresponding genes were later identified in sequences originating from chromosomes 19 and 7, respectively. On the basis of the collected information from gene and cDNA sequences, Hkr18 was found to be a protein of 94 kDa with 20 zinc finger motifs in its C terminus. The HKr19 is a smaller protein, with a molecular weight of 56 kDa containing 11 zinc finger motifs. Both HKr18 and HKr19 contained a KRAB A as well as a KRAB B domain in their N termini. Northern blot analysis showed expression of HKr18 in all human tissues tested, indicating a ubiquitous expression pattern. In contrast, HKr19 showed a more restricted tissue distribution, with detectable expression primarily in testis and fetal tissues. The HKr19 protein is a member of the large ZNF91 subfamily of KRAB zinc finger genes. A PCR-based analysis of the expression of HKr19 and other closely related genes showed that lymphoid, myeloid, and nonhematopoietic cells expressed different sets of these genes. This latter finding indicates that some members of the ZNF91 family may be involved in regulating lineage commitment during hematopoietic development. Transfection of various parts of HKr19 into human embryonic kidney cells (HEK 293 cells) showed that the entire protein and its zinc finger region were toxic to these cells when expressed at high levels. In contrast, the KRAB domain and the linker region seemed to be well tolerated. PMID- 11410165 TI - Structural and functional characterization of the human gene for sorting nexin 1 (SNX1). AB - The aim of the present study was to identify the gene for sorting nexin 1 (SNX1) to evaluate the potential for tissue-specific alternative splicing and to analyze the activity of the SNX1 promoter. The coding DNA for SNX1 was divided between 15 exons in 43 kb of genomic DNA located on human chromosome 15q22. Although SNX1 mRNA expression was widespread in human tissues, alternative splicing is thought to generate skipped exons in SNX1 cDNAs. By determination of the SNX1 gene structure and an analysis of the mRNAs in a variety of tissues using RT-PCR, we demonstrated that SNX1 mRNAs are alternatively spliced. Exon-skipped products were less abundant than full-length SNX1 mRNA species, but the ratio of skipped to full-length mRNA indicated that alternative splicing may be developmentally regulated in the liver. Consistent with widespread mRNA expression, the SNX1 promoter was GC rich and lacked a TATA box, features characteristic of housekeeping promoters. The promoter activity was dependent on the presence of proximal sequences that contained initiator elements and predicted binding sites for the transcription factors Sp1 and E2F. These findings indicate that regulation of SNX1 gene expression at the transcriptional level is likely minor. Rather, developmentally specific exon skipping provides a potential mechanism for regulating the activity of SNX1. PMID- 11410166 TI - cDNA transfection of amino-terminal fragment of urokinase efficiently inhibits cancer cell invasion and metastasis. AB - Focusing of urokinase-type plasminogen activator (uPA) to the cell surface via binding to its specific receptor (uPAR, CD87) is critical for tumor invasion and metastasis. Consequently, the inhibition of uPA-uPAR interaction on the cell surface might be a promising anti-invasion and anti-metastasis strategy. We examined the effects of cDNA transfection of the human uPA amino-terminal fragment (ATF) on invasion and metastasis of cancer cells. First, a highly metastatic human lung giant-cell carcinoma cell line (PG), used as the target cell for evaluation of this effect, was demonstrated to express both uPA and uPAR. Then, ATF, which contains an intact uPAR binding site but is catalytically inactive, was designed as an antagonist of uPA-uPAR interaction and was transfected into PG cells. [(3)H]-Thymidine incorporation and cell growth curves indicated that expressed ATF did not affect the proliferation of transfected cells. However, analysis by scanning electron microscopy revealed that ATF changed the host cells from the typical invasive phenotype to a noninvasive one. Correspondingly, the modified Boyden chamber test in vitro showed that ATF expression significantly decreased the invasive capacity of transfected cells. Furthermore, in the spontaneous metastasis model, it was confirmed in vivo that expressed ATF remarkably inhibited lung metastasis of implanted ATF-transfected PG cells. In summary, autocrine ATF could act as an antagonist of uPA-uPAR interaction, and ATF cDNA transfection could efficiently inhibit the invasion and metastasis of the cancer cells. Inhibition of uPA-uPAR interaction on the cell surface might be a promising anti-invasion and anti-metastasis strategy. PMID- 11410168 TI - Forces and moments telemetered from two distal femoral replacements during various activities. AB - Two distal femoral replacements were instrumented to measure axial force, torque and bending moments in the prosthesis shaft. Data are reported up to 2.5 years for the following activities: uni- and bi-lateral standing, walking, stair climbing and descending, treadmill walking, jogging and jumping. In the first subject the greatest averaged peak shaft forces found were: jogging 3.6Bodyweight (BW), stair descending 3.1BW, walking 2.8BW, treadmill walking 2.75BW, and stair ascending 2.8BW. Bending moments about the antero-posterior axis (varus-valgus) and medio-lateral axis (flexion-extension) peaked in the range 8.5-9.8 and 4.7 7.6BWcm respectively, over the follow-up period. Axial torques peaked in the range 0.2-1.3BWcm, outwardly directed. At most follow-up sessions, forces and moments during jogging were generally greater than those for other gait activities. In the second subject forces and moments were generally only 45-70% of those in the first subject, due to inadequate musculature around the knee. The data can be applied to the design and testing of distal femoral replacements and even to total knee replacements, and contributes to the knowledge of forces acting in the distal femur during activity. PMID- 11410169 TI - Loads acting in an intramedullary nail during fracture healing in the human femur. AB - The form and function of the musculo-skeletal system is closely related to the forces acting in its components. Significant forces are present in the long bones, but their magnitudes have so far only been estimated from mathematical models. Fracture fixation by means of metal implants provides an opportunity to measure the implant-born forces and to estimate the long bone forces before healing occurs. The load changes during fracture healing may provide additional information. Therefore, a telemetrized, interlocking femoral nail for wireless transmission of forces and moments acting across the fracture site was developed. The design was based on the geometry and material of a 16 mm AO nail with a circular, closed cross-section allowing full protection of the electronic circuits from the body fluids. After careful testing, it was implanted in a 33 year-old patient who had sustained a multifragmentary fracture of the left femur. Measurements at a rate of approx. 0.4 Hz were performed in different patient postures between the 2nd and 26th postoperative week. Significant axial forces and bending moments were measured during several activities such as sitting, unsupported leg elevation and partial weight bearing in a standing position. Forces orthogonal to the nail axis remained small. The reductions of the implant loads due to fracture consolidation were in the order of 50%. Dynamization of the nail did not change the forces. Even though the telemetry system did not allow for dynamic measurements and the results presented here provide data from one subject only, the new information will be useful with respect to implant design, biomechanics of fracture fixation and evaluation of healing progression. PMID- 11410170 TI - Hip contact forces and gait patterns from routine activities. AB - In vivo loads acting at the hip joint have so far only been measured in few patients and without detailed documentation of gait data. Such information is required to test and improve wear, strength and fixation stability of hip implants. Measurements of hip contact forces with instrumented implants and synchronous analyses of gait patterns and ground reaction forces were performed in four patients during the most frequent activities of daily living. From the individual data sets an average was calculated. The paper focuses on the loading of the femoral implant component but complete data are additionally stored on an associated compact disc. It contains complete gait and hip contact force data as well as calculated muscle activities during walking and stair climbing and the frequencies of daily activities observed in hip patients. The mechanical loading and function of the hip joint and proximal femur is thereby completely documented. The average patient loaded his hip joint with 238% BW (percent of body weight) when walking at about 4 km/h and with slightly less when standing on one leg. This is below the levels previously reported for two other patients (Bergmann et al., Clinical Biomechanics 26 (1993) 969-990). When climbing upstairs the joint contact force is 251% BW which is less than 260% BW when going downstairs. Inwards torsion of the implant is probably critical for the stem fixation. On average it is 23% larger when going upstairs than during normal level walking. The inter- and intra-individual variations during stair climbing are large and the highest torque values are 83% larger than during normal walking. Because the hip joint loading during all other common activities of most hip patients are comparably small (except during stumbling), implants should mainly be tested with loading conditions that mimic walking and stair climbing. PMID- 11410171 TI - Duration and frequency of every day activities in total hip patients. AB - Little knowledge about frequency and duration of daily activities in patients after total hip arthroplasty is available. Such information is required for the definition of realistic load collectives for pre-clinical testing of prostheses. It could also be helpful for the quantitative evaluation of surgery outcome with different prosthesis types. The purpose of this study was to develop and apply a system for the determination of frequency and duration of patient activities in their habitual environment and to compare the results to a clinical outcome score (Harris hip score).A portable activity monitoring system (weight 1.6 kg including batteries) was designed using a Palm top computer, 2 inclination sensors for the thigh and calf and one goniometer positioned at the knee joint. An algorithm was developed to identify frequency and duration of the activities lying, sitting, standing, walking and stair climbing from the signals of the 3 sensors. 42 patients participated in the study and were equipped with the system in the morning at their home. Datasets of 31 patients (age 62.5+/-11.5 y) covered more than 6h (9.8 +/- 1.6 h) and were included in the analysis. Prosthesis specific data as well as the Harris hip score were collected. The most frequent patient activity was sitting (44.3% of the time), followed by standing (24.5%), walking (10.2%), lying (5.8%) and stair climbing (0.4%). The median number of steps/stairs was 6048/164. The number of step cycles representing one year in vivo use should, consequently, be increased to 1.1 million. The Harris hip score (91.4 +/- 9.8) correlated significantly with the number of stairs (r(2) = 0.26, p = 0.003) and showed a positive tendency with the number of steps per day. No differences in activity levels between prosthesis specific factors were found. PMID- 11410172 TI - Musculo-skeletal loading conditions at the hip during walking and stair climbing. AB - Musculo-skeletal loading plays an important role in the primary stability of joint replacements and in the biological processes involved in fracture healing. However, current knowledge of musculo-skeletal loading is still limited. In the past, a number of musculo-skeletal models have been developed to estimate loading conditions at the hip. So far, a cycle-to-cycle validation of predicted musculo skeletal loading by in vivo measurements has not been possible. The aim of this study was to determine the musculo-skeletal loading conditions during walking and climbing stairs for a number of patients and compare these findings to in vivo data. Following total hip arthroplasty, four patients underwent gait analysis during walking and stair climbing. An instrumented femoral prosthesis enabled simultaneous measurement of in vivo hip contact forces. On the basis of CT and X ray data, individual musculo-skeletal models of the lower extremity were developed for each patient. Muscle and joint contact forces were calculated using an optimization algorithm. The calculated peak hip contact forces both over- and under-estimated the measured forces. They differed by a mean of 12% during walking and 14% during stair climbing. For the first time, a cycle-to-cycle validation of predicted musculo-skeletal loading was possible for walking and climbing stairs in several patients. In all cases, the comparison of in vivo measured and calculated hip contact forces showed good agreement.Thus, the authors consider the presented approach as a useful means to determine valid conditions for the analysis of prosthesis loading, bone modeling or remodeling processes around implants and fracture stability following internal fixation. PMID- 11410173 TI - Normal hip joint contact pressure distribution in single-leg standing--effect of gender and anatomic parameters. AB - A practical and easy-to-use analysis technique that can study the patient's hip joint contact force/pressure distribution would be useful to assess the effect of abnormal biomechanical conditions and anatomical deformities on joint contact stress for treatment planning purpose. This technique can also help to establish the normative database on hip joint contact pressure distribution in men and women in different age groups. Twelve anatomic parameters and seven biomechanical parameters of the hip joint in a normal population (41 females, 15 males) were calculated. The inter-parameter correlations were investigated. The pressure distribution in the hip joint was calculated using a three-dimensional discrete element analysis (DEA) technique. The 3D contact geometry of the hip joint was estimated from a 2D radiograph by assuming that the femoral head and the acetabular surface were spherical in shape. The head-trochanter ratio (HT), femoral head radius, pelvic height, the joint contact area, the normalized peak contact pressure, abductor force, and the joint contact force were significantly different between men and women. The normalized peak contact pressure was correlated both with acetabular coverage and head-trochanter ratio. Change of abductor force direction within normal variation did not affect the joint peak contact pressure. However, in simulated dysplastic conditions when the CE angle is small or negative, abductor muscle direction becomes very sensitive in joint contact pressure estimation. The models and the results presented can be used as the reference base in computer simulation for preoperative planning in pelvic or femoral osteotomy. PMID- 11410174 TI - Gait characteristics of patients with knee osteoarthritis. AB - The knee kinematics and kinetics of 139 patients (47 males and 92 females) with Grade II knee osteoarthritis (OA) were measured during level walking, stair ascent and stair descent. There was no significant difference in knee motion between the patients and normal subjects. The patients with knee OA had a significantly reduced internal knee extensor moment compared to normal subjects. This difference reflects the patient's compensation to reduce the knee joint loading. Further, subjects with OA and a higher body mass index have a lower knee extensor moment. The female subjects had significantly greater knee flexion and a greater knee extensor moment. This gender difference may partially explain the increased prevalence of OA in females. Most tests of OA treatments are assessed by criteria that do not reflect functional activities. This study demonstrates that objective gait analysis can be used to document gait adaptations used by patients with knee OA. PMID- 11410175 TI - Hip-joint and abductor-muscle forces adequately represent in vivo loading of a cemented total hip reconstruction. AB - Using finite element analyses, we investigated which muscle groups acting around the hip-joint most prominently affected the load distributions in cemented total hip reconstructions with a bonded and debonded femoral stem. The purpose was to determine which muscle groups should be included in pre-clinical tests, predicting bone adaptation and mechanical failure of cemented reconstructions, ensuring an adequate representation of in vivo loading of the reconstruction. Loads were applied as occurring during heel-strike, mid-stance and push-off phases of gait. The stress/strain distributions within the reconstruction, produced by the hip-joint contact force, were compared to ones produced after sequentially including the abductors, the iliotibial tract and the adductors and vastii. Inclusion of the abductors had the most pronounced effect. They neutralized lateral bending of the reconstruction at heel-strike and increased medial bending at mid-stance and push-off. Bone strains and stem stresses were changed accordingly. Peak tensile cement stresses were reduced during all gait phases by amounts up to 50% around a bonded stem and 11% around a debonded one. Additional inclusion of the iliotibial tract, the adductors and the vastii produced relatively small effects during all gait phases. Their most prominent effect was a slight reduction of bone strains at the level of the stem tip during heel-strike. These results suggest that a loading configuration including the hip joint contact force and the abductor forces can adequately reproduce in vivo loading of cemented total hip reconstructions in pre-clinical tests. PMID- 11410176 TI - Intermuscular interaction via myofascial force transmission: effects of tibialis anterior and extensor hallucis longus length on force transmission from rat extensor digitorum longus muscle. AB - Force transmission in rat anterior crural compartment, containing tibialis anterior (TA), extensor hallucis longus (EHL) and extensor digitorum longus (EDL) muscles, was investigated. These muscles together with the muscles of the peroneal compartment were excited maximally. Force was measured at both proximal and distal tendons of EDL muscle as well as at the tied distal tendons of TA and EHL muscles (the TA + EHL complex). Effects of TA + EHL complex length and force on proximally and distally measured forces of EDL muscle kept at constant muscle tendon complex length were assessed. Length changes of EDL muscle were imposed by movement of the proximal force transducer to different positions.Proximal EDL force was unequal to distal EDL force (active as well as passive) over a wide range of EDL muscle-tendon complex lengths. This is an indication that force is also transmitted out of EDL muscle via pathways other than the tendons (i.e. inter- and/or extramuscular myofascial force transmission). At constant low EDL length, distal lengthening of the TA + EHL complex increased proximal EDL force and decreased distal EDL force. At optimum EDL length, TA+EHL active force was linearly related to the difference between proximal and distal EDL active force. These results indicate intermuscular myofascial force transmission between EDL muscle and the TA + EHL complex. The most likely pathway for this transmission is via connections of the intact intermuscular connective tissue network. The length effects of the TA + EHL complex can be understood on the basis of changes in the configuration, and consequently the stiffness, of these connections. Damage to connective tissue of the compartment decreased the proximo-distal EDL force difference, which indicates the importance of an intact connective tissue network for force transmission from muscle fibers to bone. PMID- 11410177 TI - Combination of reduced oxygen tension and intermittent hydrostatic pressure: a useful tool in articular cartilage tissue engineering. AB - Cartilage cells are normally studied under atmospheric pressure conditions and without loading. However, since cartilage exists in a condition of reduced oxygen and intermittent hydrostatic pressure we hypothesized lower partial oxygen pressures (PO2) and different intermittent hydrostatic pressures (IHP) would increase articular chondrocyte proliferation and matrix production and to stabilize chondrocyte phenotype in vitro. Monolayers of adult bovine articular chondrocytes were cultured under 5% or 21% PO2 in combination with IHP (0.2 MPa amplitude, frequencies 5/5s = 0.1 Hz, 30/2 or 2/30 min on/off loading). We measured proliferation (3H-thymidine incorporation) and collagen secretion (protein-binding assay, collagen type II-ELISA and immunocytochemical staining of pericellular collagen types I, II and IX). Reduced PO2 stimulated proliferation and collagen type II and IX secretion of chondrocytes in comparison to 21% PO2. Additionally, collagen type I expression was delayed by low PO2, indicating a stabilization of the cell phenotype. IHP 5/5s and 30/2 min inhibited proliferation but increased collagen secretion (pericellular collagen type IX was decreased). IHP 30/2 min delayed first expression of collagen type I. In contrast, IHP 2/30 min increased proliferation, but lowered collagen expression. All stimulating or inhibiting effects of PO2 and IHP were additive and vice versa. Reduced PO2 and different settings of IHP increased proliferation, collagen secretion, and phenotype stability of chondrocytes. The oxygen- and IHP induced effects were additive, suggesting that a combination of these parameters might be a useful tool in cartilage tissue engineering. PMID- 11410178 TI - Limitations of Doppler echocardiography for the post-operative evaluation of aortic coarctation. AB - Doppler blood flow measurements and derived pressure differences, through the Bernoulli equation, are used in the diagnosis of aortic coarctation, a congenital stenosis distal to the left subclavian artery. Doppler velocities remain elevated at the coarctation site after successful repair of coarctation, leading to high Doppler derived pressure differences without significant arm-leg pressure differences. We studied this apparent contradiction of two diagnostic methods, in vivo using patient and control data, and in vitro using a hydraulic model. Clinical and echocardiographic data from 31 patients, aged 13.0 +/- 4.0, 10.5 +/- 4.7 yr after coarctectomy by end-to-end anastomosis, and 18 age-matched healthy subjects were reviewed. Doppler peak velocities at the aortic isthmus were elevated in patients (2.2 +/- 0.4 vs. 1.2 +/- 0.2m/s, P < 0.001), corresponding to significant Doppler differences (20 +/- 7 mmHg), however, without significant arm-leg pressure differences. In all patients, a mild anatomic stenosis could still be observed. Local stiffness was increased. The hypothesis that the less distensible surgical scar in post-coarctectomy patients leads to a significant dynamic obstruction in systole was validated in a latex model of the aorta. Rigid rings (0.5-1.5 cm), matching the unloaded aortic diameter, were mounted around the aorta. Under loading conditions, Doppler peak velocities increased by 40 +/ 7%, yielding Doppler differences of 21 +/- 3 mmHg, without a significant pressure drop. An alternative expression to calculate pressure differences, using both velocity and geometric information, was validated in the model. In conclusion, post-operatively, Doppler velocities remain elevated due to a mild anatomical and significant dynamic narrowing, but the specific geometry, resembling a tubular hypoplasia rather than an abrupt stenosis, permits an almost complete pressure recovery explaining the occurrence of Doppler differences in disagreement with the negligible arm-leg pressure difference. PMID- 11410179 TI - The fate of mechanically induced cartilage in an unloaded environment. AB - According to mechanobiologic theories, persistent intermittent mechanical stimulation is required to maintain differentiated cartilage. In a rat model for bone repair, we studied the fate of mechanically induced cartilage after unloading. In three groups of rats, regenerating mesenchymal tissue was submitted to different loading conditions in bone chambers. Two groups were immediately killed after loading periods of 3 or 6 weeks (the 3-group and the 6-group). The third group was loaded for 3 weeks and then kept unloaded for another 3 weeks (the (3 + 3)-group). Cartilage was found in all loaded groups. Without loading, cartilage does not appear in this model. In the 3-group there was no clear ongoing endochondral ossification, the 6-group showed ossification in 2 out of 5 cartilage containing specimens, and in the (3 + 3)-group all cartilage was undergoing ossification. These results suggest a tendency of the cartilage to be maintained also under unloaded conditions until it is reached by bone that can replace it through endochondral ossification.Additional measurements showed less amount of new bone in the loaded specimens. In most of the loaded specimens in the 3-group, necrotic bone fragments were seen embedded in the fibrous tissue layer close to the loading piston, indicating that bone tissue had been resorbed due to the hydrostatic compressive load. In some specimens, a continuous cartilage layer covered the end of the specimen and seemed to protect the underlying bone from pressure-induced resorption. We suggest that one of the functions of the cartilage forming in the compressive loaded parts of a bone callus is to protect the surrounding bone callus from pressure-induced fluid flow leading to resorption. PMID- 11410180 TI - Accuracy of the functional method of hip joint center location: effects of limited motion and varied implementation. AB - Accurate location of the hip joint center is essential for computation of hip kinematics and kinetics as well as for determination of the moment arms of muscles crossing the hip. The functional method of hip joint center location involves fitting a pelvis-fixed sphere to the path traced by a thigh-fixed point while a subject performs hip motions; the center of this sphere is the hip joint center. The aim of the present study was to evaluate the potential accuracy of the functional method and the dependence of its accuracy on variations in its implementation and the amount of available hip motion. The motions of a mechanical linkage were studied to isolate the factors of interest, removing errors due to skin movement and the palpation of bony landmarks that are always present in human studies. It was found that reducing the range of hip motion from 30 degrees to 15 degrees did significantly increase hip joint center location errors, but that restricting motion to a single plane did not. The magnitudes of these errors, however, even in the least accurate cases, were smaller than those previously reported for either the functional method or other methods based on pelvis measurements of living subjects and cadaver specimens. Neither increasing the number of motion data observations nor analyzing the motion of a single thigh marker (rather than the centroid of multiple markers) was found to significantly increase error. The results of this study (1) imply that the limited range of motion that is often evident in subjects with hip pathology does not preclude accurate determination of the hip joint center when the functional method is used; and (2) provide guidelines for the use of the functional method in human subjects. PMID- 11410181 TI - Time for creative thinking about men's health. PMID- 11410182 TI - Lessons from the arterial-switch operation. PMID- 11410183 TI - Postmarketing surveillance study of Norplant in developing countries. PMID- 11410184 TI - Natural history of cervical human papillomavirus infections. PMID- 11410185 TI - Cost-effectiveness of the implantable cardioverter defibrillator? PMID- 11410186 TI - Combs, ducks, and the brain. PMID- 11410187 TI - Review of clinical protocols at The Lancet. PMID- 11410188 TI - The Lancet--now less sharp. PMID- 11410189 TI - Are all wheezing disorders in very young (preschool) children increasing in prevalence? AB - BACKGROUND: Distinct wheezing disorders co-exist in young (preschool) children, some of which (early transient wheeze and viral wheeze) are thought to be unrelated to atopy. Investigation of changes in prevalence of wheezing disorders in preschool children could provide important clues about underlying mechanisms responsible for increasing prevalence of asthma in schoolchildren. METHODS: Repeated population surveys of the prevalence of respiratory symptoms were done by parent-completed postal questionnaires in random samples of 1650 (1990) and 2600 (1998) caucasian children aged 1-5 years living in the county of Leicestershire, UK. FINDINGS: The response rates were 86% (1422 of 1650) in 1990 and 84% (2127 of 2522) in 1998. Between 1990 and 1998, there was a significant increase in the prevalance of reported wheeze ever (16% to 29%, p<0.0001), current wheeze (12% to 26%, p<0.0001), diagnosis of asthma (11% to 19%, p<0.0001), treatment for wheeze (15% to 26%, p<0.0001), and admission for wheeze or other chest trouble (6% to 10%, p<0.0001). The increase occurred both in children with viral wheeze (9% to 19%) and in those with the classic asthma pattern of wheezing with multiple triggers (6% to 10%). There was also an increase in transient early wheezers (3% to 5%), persistent wheezers (5% to 13%), and late-onset wheezers (6% to 8%), and in all severity groups. The increase could not be accounted for by putative household risk factors because these declined between the 2 years. INTERPRETATION: The fact that all preschool wheezing disorders increased (including viral wheeze) makes it probable that factors unrelated to atopy are implicated in the changing epidemiology of wheeze in childhood. PMID- 11410190 TI - Results of the arterial switch operation in neonates with transposed great arteries. AB - BACKGROUND: The arterial switch operation is judged the best palliative operation for neonates with transposed great arteries. We aimed to assess the value of analysing a large series of unselected cases by this technically demanding operation and formulate a realistic prognosis. METHODS: We reviewed all 432 neonates (mean age at operation 7 days, mean weight 3.25 kg) who underwent an arterial switch operation between 1987 and 1999. Follow-up (mean time: 4.9 years) was complete in 412 patients. FINDINGS: Survival probability and freedom from reoperation was 94% and 78% at 10 years, respectively. 26 patients died, 16 because of myocardial ischaemia. Risk factors for death included early experience, low weight, associated cardiovascular malformations (especially hypoplasia of the right ventricle or aortic arch), and difficult patterns of coronary arteries. The risk of the coronary artery pattern was greatly reduced in those who had recent operations. At last follow-up, 90% of patients had normal life without treatment, and 94% a normal heart function on echocardiography. INTERPRETATION: The arterial switch operation in neonates achieves excellent results mid-term. Obstruction of the translocated coronary arteries is responsible for most deaths and a substantial number of reoperations. Although confirmation is needed, these results allow anticipation of a favourable long term prognosis. PMID- 11410191 TI - Natural history of cervical human papillomavirus infection in young women: a longitudinal cohort study. AB - BACKGROUND: Laboratory and epidemiological research suggests an association between human papillomavirus (HPV) and cervical intraepithelial neoplasia (CIN). We studied the natural history of incident cervical HPV infection and its relation to the development of CIN. METHODS: We recruited 2011 women aged 15-19 years who had recently become sexually active. We took a cervical smear every 6 months and stored samples for virological analysis. We immediately referred all women with any cytological abnormality for colposcopic assessment, but postponed treatment until there was histological evidence of progression to high-grade CIN. FINDINGS: In 1075 women who were cytologically normal and HPV negative at recruitment, the cumulative risk at 3 years of any HPV infection was 44% (95% CI 40-48): HPV 16 was the most common type. The cumulative risk at 3 years of detecting an HPV type not present in the first positive sample was 26% (20-32). 246 women had an abnormal smear during follow-up, of whom 28 progressed to high grade CIN. The risk of high-grade CIN was greatest in women who tested positive for HPV 16 (risk ratio 8.5 [3.7-19.2]); this risk was maximum 6-12 months after first detection of HPV 16. All HPV types under consideration were associated with cytologically abnormal smears. Although abnormality was significantly less likely to be associated with low-viral-load samples, the cumulative risk at 3 years of a high-viral-load sample after a low-viral-load sample was 45% (95% CI 35-56). Five women who progressed to high-grade CIN consistently tested negative for HPV. INTERPRETATION: Our findings suggest that attempts to exploit the association between cervical neoplasia and HPV infection to improve effectiveness of cervical screening programmes might be undermined by the limited inferences that can be drawn from the characterisation of a woman's HPV status at a single point in time, and the short lead time gained by its detection. PMID- 11410192 TI - Control of malaria in Pakistan by applying deltamethrin insecticide to cattle: a community-randomised trial. AB - BACKGROUND: The standard method of malaria control in south Asia, indoor spraying of houses with residual insecticide, is becoming prohibitively expensive to implement and new approaches are needed. Since the region's vector mosquitoes feed predominantly on domestic animals and only secondarily on human beings, to apply insecticide to surfaces of cattle instead might be more costeffective. We aimed to investigate whether domestic livestock treated with deltamethrin (applied by a sponging method) would prove toxic to mosquitoes and therefore aid in malaria control. METHODS: Six Afghan refugee settlements in Pakistan were randomly assigned to one of two groups. In one group livestock were treated with deltamethrin during the malaria transmission seasons of 1995 and 1997, whereas in the other group livestock were treated during the 1996 season. Malaria was monitored by passive case detection at village clinics and by cross-sectional surveillance. Mosquitoes were also monitored. FINDINGS: According to clinic records the incidence of malaria caused by Plasmodium falciparum decreased by 56% (95% CI 14-78%) and P vivax by 31% (5-50%) in livestock-treated villages. Cross sectional surveys showed comparable decreases in parasite prevalence. The density and life expectancy of Anopheles stephensi and A culicifacies populations were reduced in treated villages. The efficacy of livestock treatment was similar to that of indoor spraying but campaign costs were 80% less. When applied in a highly endemic settlement, the incidence of falciparum malaria decreased from 280 episodes per 1000 person-years to nine episodes per 1000 person-years. INTERPRETATIONS: Insecticide treatment of livestock is a cost-effective and promising alternative for south Asia and other regions where primary vectors are zoophilic. PMID- 11410193 TI - Efficacy and safety of infliximab monotherapy for plaque-type psoriasis: a randomised trial. AB - BACKGROUND: Currently available treatments for moderate to severe psoriasis are either incompletely effective in some patients, or are associated with toxic effects. Since tumour necrosis factor alpha (TNF-alpha) is thought to have a role in the pathogenesis of psoriasis, we did a double-blind, randomised trial to assess the clinical benefit and safety of infliximab-a monoclonal antibody against TNF-alpha. METHODS: 33 patients with moderate to severe plaque psoriasis were randomly assigned intravenous placebo (n=11), infliximab 5 mg/kg (n=11), or infliximab 10 mg/kg (n=11) at weeks 0, 2, and 6. Patients were assessed at week 10 for the primary endpoint (score on the physician's global assessment [PGA]). Analysis was by intention to treat. FINDINGS: Of the 33 patients enrolled, three dropped out. Nine of 11 (82%) patients in the infliximab 5 mg/kg group were responders (good, excellent, or clear rating on PGA), compared with two of 11 (18%) in the placebo group (difference 64% [95% CI 20-89], p=0.0089), and ten of 11 (91%) patients in the infliximab 10 mg/kg group were responders (difference from placebo 73% [30-94], p=0.0019). The median time to response was 4 weeks for patients in both infliximab groups. There were no serious adverse events, and infliximab was well tolerated. INTERPRETATION: In this controlled trial, patients receiving the anti-TNF-alpha agent infliximab as monotherapy experienced a high degree of clinical benefit and rapid time to response in the treatment of moderate to severe plaque psoriasis compared with patients who received placebo. These findings suggest that TNF-alpha has a pivotal role in the pathogenesis of psoriasis. PMID- 11410194 TI - A traumatic car crash. PMID- 11410195 TI - Detection of prostate cancer with a microbubble ultrasound contrast agent. AB - The diagnosis of prostate cancer is currently limited by the low sensitivity and specificity of systematic conventional grey-scale ultrasonography. We assessed contrast-enhanced colour Doppler ultrasonography by means of a microbubble ultrasound contrast agent to detect tumour vascularity and improve the diagnosis of prostate cancer. The use of a microbubble ultrasound contrast agent for transrectal colour Doppler targeted biopsy significantly improved the detection of prostate cancer compared with systematic biopsy following conventional grey scale ultrasonography (p<0.001). Contrast-agent enhanced colour Doppler imaging may allow for limited targeted biopsies (five or less), which reduces costs and morbidity. PMID- 11410196 TI - Anti-epiligrin cicatricial pemphigoid and relative risk for cancer. AB - It is not known whether patients with anti-epiligrin cicatricial pemphigoid (AECP) have an increased risk of malignancy. We calculated the expected numbers of cancers in a cohort of 35 such patients based on respective incidence rates for all cancers in the National Cancer Institute's Surveillance, Epidemiology, and End Results (NCI SEER) Registry. Ten patients in this cohort had solitary solid cancers; eight patients developed cancer after onset of AECP (seven within 14 months). The relative risk (RR) for cancer in this cohort was 6.8 (95% confidence intervals [CI]: 3.3-12.5). AECP seems to be associated with an increased relative risk for cancer. PMID- 11410197 TI - Variation in antibiotic use in the European Union. AB - Data on antibiotic use are not publicly available in most European Union countries. We obtained data for non-hospital antibiotic sales for 1997 from the 15 member states and analysed these according to the Anatomic Therapeutic Chemical classification system, and expressed them as defined daily doses per 1000 people per day. Sales of antibiotics varied more than four-fold: France (36.5), Spain (32.4), Portugal (28.8), and Belgium (26.7) had the highest sales, whereas the Netherlands (8.9), Denmark (11.3), Sweden (13.5), and Germany (13.6) had the lowest. There was also profound variation in use of different classes of antibiotics. Detailed knowledge of antibiotic use is necessary to implement national strategies for optimum antibiotic use, and to address the threat posed by resistant microorganisms. PMID- 11410198 TI - Western and eastern European trends in testicular cancer mortality. AB - Testicular cancer is curable if treated appropriately. We used national mortality data to compare specific death rates from the disorder in western and eastern Europe, the USA, and Japan. Testicular cancer mortality rates have fallen by about 70% in the USA and western Europe since the 1970s. In eastern Europe, however, death rates from testicular cancer have been declining only since the late 1980s, and at a much slower rate than that recorded elsewhere (about 20%). Consequently, many avoidable deaths, mainly in young adults, are still occurring in eastern Europe. Available effective treatment strategies for testicular cancer must be implemented in these countries. PMID- 11410199 TI - US government warns practitioner database underused. PMID- 11410200 TI - VEGF implicated in degeneration of motor neurons. PMID- 11410201 TI - Paediatric heart sounds assessed by computer. PMID- 11410202 TI - A possible molecular explanation for pre-eclampsia. PMID- 11410205 TI - Field experience guides PAHO's vaccine director. PMID- 11410206 TI - Accra. A way forward for mental health care in Ghana? PMID- 11410207 TI - Gloomy anniversary and outlook for HIV/AIDS. PMID- 11410208 TI - Death of child AIDS activist marks little change in South Africa. PMID- 11410209 TI - Amnesty report highlights widespread global human rights abuses. PMID- 11410210 TI - Canadian epidemiologists examine elective surgery. PMID- 11410211 TI - Over-the-counter sale of antimalaria drugs stalls Kenyan disease strategy. PMID- 11410212 TI - New psychiatric centre in Israel tackles "society wide" stress. PMID- 11410213 TI - Primary immunodeficiency diseases: an experimental model for molecular medicine. AB - Primary immunodeficiency diseases represent a vast array of inherited disorders of the immune system. Major advances in the understanding of genetic basis and molecular mechanisms have occurred within the past 10 years, as a result of the tools of modern genetics. About three quarters of 100 primary immunodeficiency diseases can now be reliably diagnosed with molecular probes. In many cases, gene identification has enabled significant insight into the physiopathology of the related conditions. Therapeutic progress based on protein engineering and possibly gene therapy will also ensue. PMID- 11410214 TI - Lessons from the glitazones: a story of drug development. AB - Troglitazone, the first in the thiazolidinedione class of oral hypoglycaemic agents, was launched in the USA in March, 1997. It reached Europe later that year, only to be withdrawn within weeks on the grounds of liver toxicity. Meanwhile it went on to generate sales of over $2 billion in the USA, and caused at least 90 cases of liver failure (70 resulting in death or transplantation) before it was withdrawn in March, 2000. Rosiglitazone and pioglitazone reached the US market in 1999 as first-line agents to be used alone or in combination with other drugs, but in Europe the same dossiers were used one year later to apply for a limited licence as second-line agents restricted to oral combination therapy. How should we use the glitazones? And how did they achieve blockbuster status without any clear evidence of advantage over existing therapy? PMID- 11410215 TI - Why did the 5th Earl of Derby die? AB - The unexpected death of Ferdinando Stanley, 5th Earl of Derby, on April 16, 1594 was an event of major political importance in the later years of Queen Elizabeth I of England. When he had succeeded his father at the age of 38 he became head of one of the most influential families in the country. He also had a claim to the throne if Elizabeth died without naming a successor. Yet within seven months of entering into his inheritance, this previously fit man was suddenly taken ill and died a fortnight later. His death was so significant that the historian John Stow recorded his illness in great detail (Fig. 1).(1) Stow's remarkable account is compatible with a sinister interpretation of the cause. PMID- 11410226 TI - The fluidity of truth. PMID- 11410227 TI - Impact of Cu(II) and Ni(II) on a structure of chiral peptide nucleic acids having four, six and eight thymines in a peptide side chain. AB - The studies on binding ability of longer chiral peptide nucleic acids (having four, six and eight thymines in a peptide side chain) have shown that the interactions between the nucleic base rings within a ligand molecule have a critical impact on the complex stability. Thymines inserted in the peptide side chain interact with each other as well as with peptide back-bone increasing the structural organization of the cPNA molecule. The metal ion coordination to cPNA, on the other hand, induces a very specific ligand structure, which may have a basic impact on the cPNA self-recognition processes. PMID- 11410228 TI - CD spectroscopic study on the speciation and solution structure of copper(II) complexes of some tripeptides in combination with potentiometric and spectrophotometric results. AB - The combination of potentiometric, spectrophotometric and CD spectroscopic studies under the same conditions is expected to yield more reliable thermodynamic and structural information for a certain system. The matrix rank analysis of both the spectrophotometric and CD spectra series gives the necessary number of species to be taken into account in the calculations. Based on such speciation diagrams the molar spectra for individual complexes in the copper(II) Gly-Gly-Ala, Gly-Ala-Gly, Ala-Gly-Gly and Ala-Ala-Ala systems were obtained. The results allowed us to assign these spectra to two types of coordination species with the compositions CuLH(-1) and [CuLH(-2)](-). The sign and the intensity of the CD spectra are well related to the chirality and distance of the chiral center(s) in the ligand from the metal ion chromophore and the stability of the metal complexes. They are further on additive within the experimental error in both cases with respect to chiral centers: the sum of the CD spectra of the Gly Gly-Ala, Gly-Ala-Gly and Ala-Gly-Gly complexes equals those of Ala-Ala-Ala complexes. The CD spectra strongly support the coordination also of the C terminal part of the peptide in the CuLH(-1) complex of GGA, and even the deprotonation of the Ala peptide group instead of that belonging to Gly cannot be excluded. PMID- 11410229 TI - Coordination mode and reactivity of copper(II) complexes with kasugamycin. AB - Protonation and Cu(II) coordination of kasugamycin were studied by potentiometry, UV-vis, CD, EPR, 13C NMR, and 1H NMR. Mononuclear complexes with stoichiometries ranging from CuHL to CuH(-1)L were found. The aminoamidine moiety provides the coordination site in the CuHL species. The additional axial coordination of the amino nitrogen of the aminosugar ring is present in CuL. Finally, the CuH(-1)L complex is formed as a result of a deprotonation and coordination of the hydroxyl group of the inositol ring. The non-planar arrangement of the chelate rings results in the relative stabilization of a Cu(I) species. As a consequence, Cu(I) and superoxide radicals are involved in the redox mechanism of H(2)O(2) activation by the Cu(II) complex of kasugamycin. PMID- 11410230 TI - Volume and enthalpy profiles of CO binding to Fe(II) tetrakis-(4 sulfonatophenyl)porphyrin. AB - The focus of this study is to examine volume and enthalpy profiles of ligand binding associated with CO-Fe(II) tetrakis-(4-sulfonato phenyl)-porphyrin (COFe(II)4SP) in aqueous solution. Temperature dependent photothermal beam deflection was employed to probe the overall enthalpy and volume changes associated with CO-photolysis and recombination. The analysis demonstrates that ligand recombination occurs with a pseudo first order rate constant of (2.5+/ 0.2)x10(4) s(-1) (at 25 degrees C) with a corresponding volume decrease of 6+/-1 ml/mol. The activation enthalpy (DeltaH(double dagger)) and volume (DeltaV(double dagger)) change for CO recombination (determined from temperature/pressure dependent transient absorption spectroscopy) are found to be 3.9 kcal/mol and 8.2 ml/mol, respectively. These data are consistent with a mechanism in which photolysis yields a five-coordinate high spin (H(2)O)Fe(II)4SP complex that recombines in a single step to form the low spin (CO)(H(2)O)Fe(II)4SP complex. Base elimination, often associated with CO photolysis from hemes, is not observed in this system. The overall volume changes suggest a transition state with significant high spin character. Furthermore, these results demonstrate the utility of coupling photothermal techniques with variable pressure/temperature transient absorption spectroscopy to probe heme reaction dynamics. PMID- 11410231 TI - Oxygen disruption of the 2[4Fe-4S] clusters in Clostridium pasteurianum ferredoxin shown by 1H-NMR. AB - The ferredoxin from Clostridium pasteurianum, which contains two [4Fe-4S] clusters, was investigated in its oxidized and reduced states by two-dimensional (2D) (1)H-(1)H nuclear Overhauser enhancement spectroscopy (NOESY). Comparison of the data from the oxidized ferredoxin with those published previously revealed the same NOE connectivities. No previous (1)H-(1)H NOESY study of the fully reduced ferredoxin has previously been published. However, it was possible to compare our results with those of a 2D exchange spectroscopy investigation of half-reduced C. pasteurianum ferredoxin. The present results with reduced C. pasteurianum ferredoxin confirm many of the (1)H peaks and NOE interactions reported earlier, revise others, and locate resonances previously undetected. When the ferredoxin was slightly exposed to oxygen, several of the hyperfine shifted resonances were irreversibly influenced. A resonance at 34 ppm in the (1)H NMR spectra of both redox states is indicative of oxygen exposure. These results indicate the importance of keeping the ferredoxin strictly anaerobic during purification and solvent exchange. PMID- 11410232 TI - Silybin, a new iron-chelating agent. AB - Silybin, a natural occurring flavolignan isolated from the fruits of Silibum marianum, has been reported to exert antioxidant and free radical scavenging abilities. It was suggested to act also as an iron chelator. The complexation and protonation equilibria of the ferric complex of this compound have been studied by potentiometric, spectrophotometric and electrochemical techniques. The formation of the complex silybin-Ga(III) in anhydrous DMSO-d6 has been studied by 1H NMR spectroscopy. Mass spectrometry and infrared spectroscopy on silybin Fe(III) complex confirm all data obtained by 1H NMR spectroscopy. The experimental results show that silybin binds Fe(III) even at acidic pH. Different ternary complexes were observed at increasing methoxide ion concentration and their stability constants have been calculated. The results show the possible role of silybin in relation to the chelation therapy of chronic iron overload, as occurs in the treatment of Cooley's anemia. PMID- 11410233 TI - The kinetics and mechanisms of the reaction of iron(III) with gallic acid, gallic acid methyl ester and catechin. AB - The kinetics and mechanisms of the reactions of a number of pyrogallol-based ligands with iron(III) have been investigated in aqueous solution at 25 degrees C and ionic strength 0.5 M NaClO(4). Mechanisms have been proposed which account satisfactorily for the kinetic data. These are generally consistent with a mechanism in which the 1:1 complex that is formed initially when the metal reacts with the ligand subsequently decays through an electron transfer reaction. There was also some evidence for the formation of a 1:2 ligand-to-metal complex at higher pH values. The kinetics of complex formation were investigated with either the ligand or metal in pseudo-first-order excess. Rate constants for k(1) of 2.83(+/-0.09)x10(3), 1.75(+/-0.045)x10(3) and 3300(+/-200) M(-1) s(-1) and k(-1) of 20(+/-6.0), 35(+/-13) and 25+/-7.6 M(-1) s(-1) have been evaluated for the reaction of Fe(OH)(2+) with gallic acid, gallic acid methyl ester and catechin, respectively. The stability constant of each [Fe(L)](+) complex has been calculated from the kinetic data. The iron(III) assisted decomposition of the initial iron(III) complex formed was investigated. Analysis of the kinetic data yielded both the equilibrium constants for protonation of the iron(III) complexes initially formed together with the rate constants for the intramolecular electron transfers for gallic acid and gallic acid methyl ester. All of the suggested mechanisms and calculated rate constants are supported by calculations carried out using global analysis of time-dependent spectra. PMID- 11410234 TI - Aluminum speciation studies in biological fluids. Part 7. A quantitative investigation of aluminum(III)-malate complex equilibria and their potential implications for aluminum metabolism and toxicity. AB - As a nonessential element, aluminum may be toxic at both environmental and therapeutic levels, depending on ligand interactions. Dietary acids that normally occur in fruits and vegetables and commonly serve as taste enhancers are good ligands of the Al(3+) ion. Malic acid is one of these and also one of the most predominant in food and beverages. The present paper reports an examination of its potential influence on aluminum bioavailability through speciation calculations based on Al(III)-malate complex formation constants especially determined for physiological conditions. According to the results obtained, malate appears to be extremely effective in maintaining Al(OH)(3) soluble over the whole pH range of the small intestine under normal dietary conditions. In addition, two neutral Al(III)--malate complexes are formed whose percentages are maximum from very low malate levels. When aluminum is administered therapeutically as its trihydroxide, the amount of metal neutralized by malate peaks as its solubility pH range regresses to its original limits in the absence of malate. The enhancing effect of malate towards aluminum absorption is therefore virtually independent of the aluminum level in the gastrointestinal tract. The presence of phosphate in the gastrointestinal juice is expected to limit the potential influence of malate on aluminum absorption. Under normal dietary conditions, phosphate effectively reduces the fraction of aluminum neutralized by malate but without nullifying it. Aluminum phosphate is predicted to precipitate when aluminum levels are raised as with the administration of aluminum hydroxide, but a significant amount of neutral aluminum malate still remains in solution. Even therapeutic aluminum phosphate is not totally safe in the presence of malate, even at low malate concentrations. As plasma simulations predict that no compensatory effect in favor of aluminum excretion may be expected from malate, simultaneous ingestion of malic acid with any therapeutic aluminum salt should preferably be avoided. PMID- 11410235 TI - Catecholamine complexes of ruthenium-edta and their redox chemistry. AB - The electrochemical and spectroelectrochemical behavior of some neurotransmitters (dopamine and L-dopa) and their corresponding novel blue ruthenium(III)-edta complexes were investigated in aqueous solutions. At pH 7-10, the free ligand species can be electrochemically oxidized in the range of 0.1-0.6 V versus SHE, yielding primarily quinone products susceptible to pH-dependent, secondary intramolecular chemical reactions, which make the redox processes irreversible. When coordinated to the ruthenium(III)-edta complex, their electrochemical and spectroelectrochemical behavior is dramatically changed, approaching that of metal complexes with noninnocent dioxolene ligands. Reduction of the ruthenium(III) moiety proceeds reversibly above pH 9, in the region from -0.5 to 0.7 V. The oxidation process centered on the catecholate ligands becomes reversible and leads exclusively to the formation of the semiquinone species, with no evidence of complications from further reactions. These changes in the electrochemical behavior of the neurotransmitters make their cyclovoltammetric waves for reduction/oxidation more defined, favoring more precise quantitative analyses. PMID- 11410236 TI - The subsequent effect of interaction between Co(2+) and human serum albumin or bovine serum albumin. AB - A notable hysteretic effect has been observed in the interaction of Co(II) with human serum albumin (HSA) or bovine serum albumin (BSA) using UV-Visible spectrometry at physiological pH (7.43), which shows that the binding between Co(II) and HSA or BSA may induce a slow transition of HSA or BSA from the conformation of weaker affinity for Co(II) to one of stronger affinity (A-B transition). The rate constants and activation parameters of this transition were measured and are discussed. It is inferred that such a conformation transition may occur due to the binding of the first Co(II) ion with the peptide segment of N-terminal residues 1-3, which results in a 'hinged movement' of the relatively hydrophobic 'valley' in the IA subdomain. This process leads to a slow conformational transition in the albumins, makes the other binding sites of Co(II) exposed, and shows a positive cooperativity effect. The LMCT (ligand-to metal charge transition) bands of the Co(II)-HSA and Co(II)-BSA systems also show a kind of hypochromic effect featuring a dipole-dipole interaction mechanism. This phenomenon is rarely reported. PMID- 11410237 TI - Interactions of d(10) metal ions with hippuric acid and cytosine. X-ray structure of the first cadmium (II)-amino acid derivative-nucleobase ternary compound. AB - The interactions of Zn(II), Cd(II) and Hg(II) with hippuric acid (hipH) were studied and several novel compounds were synthesized and studied by NMR. Some new metal-hippuric-cytosine ternary compounds were formed and the structure of the [Cd(hip)(2)(cyt)(H(2)O)](2) ternary complex resolved. Each cadmium (II) atom has a distorted trigonal bipyramid coordination which is linked to a water molecule, a cytosine via N(3), a carboxylic oxygen atom of a hippurate moiety and two bridging dicoordinated hippurates bound through the carboxylic oxygen atoms. To these five main bonds, two longer ancillary interactions can be observed: the second oxygen of the monocoordinated hippurate group and the carboxylic oxygen of the cytosine ligand. The compound is stabilized by an intramolecular stacking between the benzene and cytosine rings and by the hydrogen bonds between the coordinated water molecules and the ligands. This is, to our knowledge, the first structure of a cadmium-amino acid derivative-natural nucleobase compound described so far. PMID- 11410238 TI - Amelioration of insulin resistance in diabetic ob/ob mice by a new type of orally active insulin-mimetic vanadyl complex: bis(1-oxy-2 pyridinethiolato)oxovanadium(IV) with VO(S(2)O(2)) coordination mode. AB - Recently, we have shown that a newly synthesized vanadyl complex, bis(1-oxy-2 pyridinethiolato)oxovanadium(IV), VO(opt)(2), is a potent orally active insulin mimetic in treating streptozotocin-induced diabetes in rats, with long-term action. In the present study, the anti-diabetic effect of VO(opt)(2) and its mechanism in ob/ob mice, an obese non-insulin-dependent diabetes mellitus (NIDDM) animal model, was investigated. In ob/ob mice, 15-day oral treatment with VO(opt)(2) resulted in a dose-dependent decrease in the levels of glucose, insulin and triglyceride in blood. VO(opt)(2) was also effective in ameliorating impaired glucose tolerance in ob/ob mice, when an oral glucose tolerance test was performed after treatment with VO(opt)(2). Tumor necrosis factor-alpha (TNF alpha) is a key component of obesity-diabetes link, we therefore examined the attenuating effect of VO(opt)(2) on impaired insulin signal transduction induced by TNF-alpha. Elevated expression of TNF-alpha was observed in the epididymal and subcutaneous fat tissues of ob/ob mice. Incubation of 3T3-L1, mouse adipocytes, with TNF-alpha reduced the phosphorylation of insulin receptor substrate-1 (IRS 1), whereas VO(opt)(2) treatment resulted in an enhancement of IRS-1 phosphorylation, irrespective of the presence or absence of TNF-alpha. Overall, the present study demonstrates that VO(opt)(2) exerts an anti-diabetic effect in ob/ob mice by ameliorating impaired glucose tolerance, and furthermore, attenuates the TNF-alpha-induced decrease in IRS-1 phosphorylation in adipocytes. These results suggest that the anti-diabetic action of VO(opt)(2) is derived from an attenuation of a TNF-alpha induced impaired insulin signal transduction via inhibition of protein tyrosine phosphatase, providing a potential clinical utility for VO(opt)(2) in the treatment of NIDDM. PMID- 11410239 TI - Metal complexes of the mycotoxins sporidesmin A and gliotoxin, investigated by electrospray ionisation mass spectrometry. AB - The mycotoxin sporidesmin A (spdA), responsible for the intoxication of animals, causing facial eczema, has been investigated by electrospray ionisation mass spectrometry. Protonated [spdA+H](+) and deprotonated [spdA-H](-) ions are observed in positive and negative ion modes respectively. Reduced spdA, formed by cleavage of the disulfide bond by Na[BH(4)] gives an ion [spdA+H](-), and forms ions of the type [2spdA+M](2-) with a range of divalent metal ions M(2+)=Zn(2+), Cd(2+), Hg(2+), Sn(2+) and Fe(2+). Sodium-containing analogues [2spdA+M+Na](-) are observed, particularly at high cone voltages, where they are stable towards cone voltage-induced fragmentation, indicating appreciable stability of the (spdA)(2)M system. A competition experiment between Cd(2+) and Zn(2+) demonstrates that reduced spdA has a higher affinity for Cd(2+) ions. The related gliotoxin (gtx) forms analogous [2gtx+M](2-) and [2gtx+M+Na](-) ions. The reduction and metal complexation of spdA can be monitored by (1)H NMR spectroscopy, and results in chemical shift changes for those protons adjacent to the sulfur atoms. The isolation of a polymeric cadmium-spdA complex is also reported. PMID- 11410240 TI - Cell death by reactive oxygen species generated from water-soluble cationic metalloporphyrins as superoxide dismutase mimics. AB - We investigated the effect on cell death of reactive oxygen species induced by water-soluble cationic metalloporphyrins with superoxide dismutase (SOD) activity. The SOD activity of 5,10,15,20-tetrakis(4-N-methylpyridyl)]porphine (MPy(4)P) containing Fe, Mn or Cu was measured using a cytochrome c assay by the xanthine/xanthine oxidase system and stopped-flow kinetic analysis. Cell viability of four cell lines treated with metalloporphyrins, mitomycin c (MMC), or cisplatin was estimated by a trypan blue exclusion assay. FeMPy(4)P with a high SOD activity showed a significant cytotoxicity compared with MMC and cisplatin, while CuMPy(4)P without SOD activity exhibited no cytotoxicity. However, MnMPy(4)P showing an SOD activity as high as that of FeMPy(4)P did not indicate cytotoxicity. These findings suggest that FeMPy(4)P as SOD mimic converts intracellular O2(*-) to H(2)O(2) and that it rapidly reacts with H(2)O(2) to form *OH, causing DNA damage and inducing cell death. On the other hand, MnMPy(4)P did not participate in the Fenton reaction, so that DNA damage in the cells treated with MnMPy(4)P was not observed. In addition, the cytotoxicity by the metalloporphyrin was inversely correlated with the SOD activity of the cells and the selective damage at cellular and DNA levels was confirmed. We believe that for an anticancer drug with antioxidant ability O(2)(*-) is useful as a target molecule to induce selective cell death between cancer and normal cells and that metalloporphyrins showing SOD activity and Fenton-like reaction are a new class of anticancer agents. PMID- 11410241 TI - cis-Dichloroplatinum(II) complexes tethered to 9-aminoacridine-4-carboxamides: synthesis and action in resistant cell lines in vitro. AB - A series of intercalator-tethered platinum(II) complexes PtLCl(2) have been prepared where L are the diamine ligands N-[2-[(aminoethyl)amino]ethyl]-9 aminoacridine-4-carboxamide, N-[3-[(2-aminoethyl)amino]propyl]-9-aminoacridine-4 carboxamide, N-[4-[(2-aminoethyl)amino]butyl]-9-aminoacridine-4-carboxamide and N [5-[(aminoethyl)amino]pentyl]-9-aminoacridine-4-carboxamide and N-[6 [(aminoethyl)amino]hexyl]-9-aminoacridine-4-carboxamide. The activity of the complexes was assessed in the CH-1, CH-1cisR, 41M, 41McisR and SKOV-3 cell lines. The compounds with the shorter linker chain lengths are generally the most active against these cell lines and are much more toxic than Pt(en)C1(2). For example, for the n=2 compound the IC(50) values are 0.017 microM (CH-1), 1.7 microM (41M), 1.4 microM (SKOV-3) and the resistance ratios are 51 (CH-1cisR) and 1.6 (41McisR). For the untethered analogue Pt(en)C1(2) the IC(50) values are 2.5 microM (CH-1), 2.9 microM (41M), 45 microM (SKOV-3) and the resistance ratios are 2.8 (CH-1cisR) and 4.1 (41McisR). The very large differential in IC(50) values between the CH-1 and CH-1cisR pair of cell lines for the 9-aminoacridine-4 carboxamide tethered platinum complexes indicates that repair of platinum-induced DNA damage may be a major determinant of the activity of these compounds. PMID- 11410242 TI - Interaction of metallopyrazoliumylporphyrins with calf thymus DNA. AB - The interaction of transition metal complexes of cationic porphyrins bearing five membered rings, meso-tetrakis(1,2-dimethylpyrazolium-4-yl)porphyrin (MPzP, M=Mn(III), Ni(II), Cu(II) or Zn(II)), with calf thymus DNA (ctDNA) has been studied. Metalloporphyrins NiPzP and CuPzP are intercalated into the 5'GC3' step of ctDNA. MnPzP is bound edge-on at the 5'TA3' step of the minor groove of ctDNA, while ZnPzP is bound face-on at the 5'TA3' step of the major groove of ctDNA. The binding constants of the metalloporphyrins to ctDNA range from 1.05x10(5) to 2.66x10(6) M(-1) and are comparable to those of other reported cationic porphyrins. The binding process of the metallopyrazoliumylporphyrins to ctDNA is endothermic and entropically driven. These results have revealed that the kind of central metal ions of metalloporphyrins influences the binding characteristics of the porphyrin to DNA. PMID- 11410243 TI - Sterically hindered cisplatin derivatives with multiple carboxylate auxiliary arms: synthesis and reactions with guanosine-5'-monophosphate and plasmid DNA. AB - Two novel sterically hindered cisplatin derivatives with the ligand L=NH(2)C(CH(2)CH(2)COOH)(3) were prepared: cis-PtCl(2)L(2) and cis PtCl(2)L(NH(3)). The starting compound for the syntheses was NH(2)C(CH(2)CH(2)COOtBu)(3), also known as a building block for dendrimers. cis PtCl(2)L(2) was prepared from K(2)PtCl(4) in an unusual two-phase reaction in water-chloroform, followed by deprotection of the tert-butyl protective groups with formic acid to yield a water-soluble complex. The mixed-ligand compound cis PtCl(2)L(NH(3)) was prepared from [PPh(4)][PtCl(3)(NH(3))] in methanol, with subsequent deprotection in formic acid. DNA-binding properties of the two compounds were investigated using the model base guanosine-5'-monophosphate (5' GMP) and pBR322 plasmid DNA. While cisplatin [cis-PtCl(2)(NH(3))(2)] induced an unwinding of 12 degrees in pBR322 plasmid DNA, cis-PtCl(2)L(NH(3)) induced only 3 degrees unwinding, which is indicative of a monofunctional binding mode. Remarkably, cis-PtCl(2)L(2) did not induce any distortion in plasmid DNA, which strongly suggests that the compound does not bind to DNA. Test reactions with 5' GMP, monitored by 1H and 195Pt NMR, confirmed that cis-PtCl(2)L(2) is unable to bind to DNA, whereas cis-PtCl(2)L(NH(3)) binds only one nucleotide. Apparently, binding of platinum to nucleotides at the coordination site cis with respect to the ligand L is prevented by steric crowding. Thus, cis-PtCl(2)L(NH(3)) must bind DNA monofunctionally at the trans position. Besides, both compounds have a chloride replaced by one of the carboxylate arms, forming a a seven-membered chelate ring. In theory, cis-PtCl(2)L(2) could also form a second chelate ring, but this was not observed. PMID- 11410244 TI - Requirement of de novo protein synthesis for aminopterin-induced apoptosis in a mouse myeloma cell line. AB - Cells synthesize nucleotides through de novo and salvage pathways that require the activities of dihydrofolate reductase (DHFR) and hypoxanthine-guanine phosphoribosyltransfease (HGPRT), respectively. Aminopterin, an inhibitor of dihydrofolate reductase, has been demonstrated to allow HGPRT(-) cells to be negatively selected. However, the pathway by which aminopterin leads to cell death remains to be clarified. In this study, we characterized features of cellular responses induced by aminopterin treatment in P3-X63-Ag8.653, a mouse HGPRT(-) myeloma cell line. Upon treatment with aminopterin, the cells readily underwent an apoptotic process, as assessed by DNA fragmentation assay and electron microscopic analysis. Aminopterin-induced apoptosis was drastically reduced by addition of actinomycin D and cycloheximide, indicating that active RNA and protein synthesis is required for the apoptotic effect of aminopterin. Interestingly, the induction of c-myc gene expression preceded the activity of DNA fragmentation in aminopterin-treated cells. Taken together, these results suggest that cells deficient in the salvage pathway of purine biosynthesis are susceptible to aminopterin-induced apoptosis that requires de novo synthesis of proapoptotic factors, including Myc oncoprotein. PMID- 11410245 TI - Expression of activation, adhesion molecules and intracellular cytokines in acute pancreatitis. AB - Adhesion and activation molecules as well as cytokines play an important role in an immune scenario. In acute pancreatitis, we have studied some of these in order to evaluate dysregulation. For this we took peripheral blood mononuclear cells and pancreatitis tissue cells. We analysed activation markers like CD69, CD25 and HLA-DR and found a marked elevation of CD69 as well as CD25 in both peripheral blood cells and tissue mononuclear cells when compared to controls. In PBMC-CD69: P<0.01 and CD25: P<0.01; in tissue-CD69: P<0.001 and CD25: P<0.001. The HLA-DR levels, however, were reduced in the disease state (in acute pancreatitis patient blood (P<0.01) and tissue cells (P<0.001)). The adhesion molecules showed unanimous rise in the blood and the tissue samples. In blood samples, CD11a: P<0.05 and CD11b: P<0.05 and tissue samples CD11a: P<0.01 and CD11b: P<0.01and CD54 in peripheral blood (P<0.05) and tissue (P<0.01) of AP was high as compared to controls. By simultaneous flowcytometric analysis, we determined the co expression of a surface marker (CD4/CD8/CD14) and intracellular cytokine (TNF alpha and IFN-gamma) in individual cells. The IFN-gamma producing CD8+T cells were elevated in pancreatic tissue (P<0.01). TNF-alpha producing cell numbers were significantly higher in tissue cells than in blood and also in CD8+ T cells (P<0.001). We conclude that monocyte function is affected in AP as shown by reduced HLA-DR numbers and lowered TNF-alpha producing cells. Moreover, the CD8+T cells appear to play an important role in cytokine synthesis at the effector site. PMID- 11410246 TI - UEA-I-binding to thymic medullary epithelial cells selectively reduces numbers of cortical TCRalphabeta+ thymocytes in FTOCs. AB - Thymic medullary epithelial cells (TMECs) constitute a major stromal cell type, the function of which is incompletely understood. Some TMECs express L-fucose glycosylated proteins on their plasma membrane; these have been shown to specifically bind the lectin UEA-I. We exploited this observation to investigate the consequences of in situ blockage of TMECs in FTOCs by UEA-I. In UEA-I-treated FTOCs, we noted a decreased cellularity among TCRalphabeta+ but not TCRgammadelta+ cells. In fact, CD3- and CD3lo cortical cells were markedly depleted, while CD3hi cells were unaffected. Since the affected cell subsets are in a different compartment from that where UEA-I binding occurs, it is likely that the effect is mediated through a soluble factor. Two possible mechanisms are proposed: a reduced activation of either TMECs or of medullary thymocytes which normally bind to them, results in lowered production of soluble factors responsible for cortical thymocyte proliferation. Alternately, the binding of UEA I to TMECs could activate the latter to produce signals inhibitory to cortical thymocytes. PMID- 11410247 TI - Differentiation by in vitro treatment of lidocaine-epinephrine and prilocaine felypressine in neutrophils. AB - Neutrophils are often the first cells of the immune system to encounter an invader, such as bacteria and fungi. Lidocaine-epinephrine induced transient potentiation of the production of superoxide anion, while prilocaine-felypressine induced persistent inhibition of the production in neutrophils. Moreover, lidocaine-epinephrine inhibited the production of hydrogen peroxide in spite that it potentiated the production of superoxide anion, while prilocaine-felypressine inhibited the production of hydrogen peroxide as well as superoxide anion. By contrast, lidocaine-epinephrine and prilocaine-felypressine are both effective in significantly inhibiting adhesion and phagocytosis. Using flow cytometric analysis, both local anesthetics were found to be effective in inhibiting the expression of Mac-1 (CD11b/CD18) in neutrophils. These results suggest that lidocaine-epinephrine and prilocaine-felypressine differentially modulate the production of superoxide anion, and could similarly inhibit adhesion, phagocytosis, and the production of hydrogen peroxide by neutrophils. PMID- 11410248 TI - Inhibition of interleukin-12 production in lipopolysaccharide-activated mouse macrophages by parthenolide, a predominant sesquiterpene lactone in Tanacetum parthenium: involvement of nuclear factor-kappaB. AB - Pharmacological control of interleukin-12 (IL-12) production may be a key therapeutic strategy for modulating immunological diseases dominated by type-1 cytokine responses. In this study, we investigated the effects of parthenolide, an anti-inflammatory sesquiterpene, on the production of IL-12 from mouse macrophages stimulated with lipopolysaccharide (LPS). Parthenolide potently inhibited the LPS-induced IL-12 production in a dose-dependent manner. The effect of parthenolide on IL-12 p40 promoter activation was analyzed by transfecting RAW264.7 monocytic cells with p40 promoter/luciferase constructs. The repressive effect mapped to a region in the p40 promoter containing a binding site for nuclear factor-kappaB (p40-kappaB). Furthermore, activation of macrophages by LPS resulted in markedly enhanced binding activity to the kappaB site, which significantly decreased upon addition of parthenolide. These results suggest that parthenolide-induced inhibition of IL-12 production in macrophages may explain some of the biological effects of parthenolide including its anti-inflammatory activity. PMID- 11410249 TI - MEK inhibitor augments IL-6-induced growth arrest and apoptosis, but antagonizes IL-6-induced macrophage differentiation of M1 cells. PMID- 11410250 TI - Immunocytochemical detection of Fcgamma receptors in human atherosclerotic lesions. AB - Much evidence indicates that atherosclerotic lesions are largely of an inflammatory nature. Activated macrophages and macrophage-derived foam cells laden with cholesterol esters are a major constituent of these lesions and can influence lesion formation via several potential mechanisms. One such mechanism is Fcgamma receptor activation and/or Fcgamma receptor-mediated clearance of immune complexes containing cholesterol, such as lipoprotein immune complexes. That this mechanism contributes to lesion formation would be further supported if Fcgamma receptor expression in arterial lesions were demonstrated. We therefore used monoclonal antibodies and immunocytochemical methods to analyze frozen sections of human arterial lesions for expression of each of the three primary classes of mononuclear phagocyte Fcgamma receptors. Approximately 800 sections of aorta, carotid, and coronary arteries obtained from five elderly donors were analyzed. The presence of macrophages was determined by assaying reactivity of a monoclonal antibody specific to CD163, which is expressed only on cells of the human mononuclear phagocyte lineage. Results indicate that highly cellular preatheromatous lesions contained numerous macrophages in the zone of proliferation that expressed each class of Fcgamma receptor (FcgammaRIA, FcgammaRIIA, and FcgammaRIIIA). Fcgamma receptor-positive cells were also present in medial and adventitial areas. Fcgamma receptor staining was both punctate and diffuse, the latter suggesting that soluble receptors were present in the extracellular matrix. These data further support that Fcgamma receptor-mediated clearance of immune complexes can occur in arterial lesions during atherogenesis. Expression of both the high affinity (FcgammaRIA) and lower affinity (FcgammaRIIA/FcgammaRIIIA) receptors indicates that mono- and multivalent IgG containing immune complexes could engage Fcgamma receptors and influence lesion formation through several different inflammatory mechanisms triggered by receptor activation. PMID- 11410251 TI - Innate immune mechanisms in the pathogenesis of systemic lupus erythematosus (SLE). AB - Immune imbalance in SLE increases the susceptibility to infectious diseases. The aim of this study was to analyze several mechanisms related to non-specific immunity in this autoimmune disorder. We studied in vivo CD11b expression, phagocytosis, and chemotaxis in polymorphonuclear cells (PMN) from SLE patients. All tests were also performed under hrIL-8 stimulating conditions and analyzed by flow cytometry. Intracellular leucocyte (monocytes and PMN) enzyme activity was evaluated using specific substrates for cathepsin B and D, collagenase, and oxidative burst by flow cytoenzymology. An exaggerated in vivo CD11b expression was observed on PMN from SLE patients without noticeably in vitro effect upon hrIL-8. Similarly both, phagocytosis and chemotaxis were diminished and showed no response to hrIL-8 stimulation. The opposite was found in PMN from controls. Intracellular enzyme activity was comparable between groups as far as cathepsin B and D are concerned. A tendency of decreased oxidative-burst induction was noted in monocytes and PMN from SLE patients, whereas collagenase activity was found clearly increased in both leucocyte subpopulations. Our results may represent a deficient ability of the innate immune mechanisms for the clearance of infectious agents, immune complexes, satisfactory resolution of inflammatory processes and tissue repair in SLE. PMID- 11410252 TI - Use of secondarily revised VH genes in IgE antibodies produced in mice infected with the nematode Nippostrongylus brasiliensis. AB - Although a high level of IgE is produced after primary infection with Nippostrongylus brasiliensis (Nb), most of the IgE antibodies (Abs) are not specific to the worm. Analyses with Western blotting and enzyme-linked immunosorbent assay (ELISA) revealed that the IgE Abs from Nb-infected BALB/c mice did not show reactivity with Nb-derived excretory-secretory proteins (NES) and antigens present in the cell-free extracts of the worm. Monoclonal IgE Abs obtained from the Nb-infected mice were not reactive with these Nb antigen either. To characterize Nb-induced IgE response, we used (QM x C57BL/6)F1 (QBF1) mice that bear the knock-in 17.2.25 VHDJH segment (VHT) encoding a VH region specific to 4-hydroxy-3-nitrophenylacetyl hapten, and express VHT-encoded antigen receptors on 80-85% of their B cells. Consistent with the frequency of VHT positive B cells, more than 80% of IgE Abs induced in QBF1 B cells that were cultured with LPS plus IL-4 were found to bear VHT-encoded H chains. In contrast, when QBF1 mice were infected with Nb, less than 10% of Nb-induced IgE Abs were found to use VHT. The QBF1-derived IgE did not react with Nb antigens either. Taken together, data suggest that Nb-induced IgE response in mice is not merely the result of polyclonal activation of B cells, but may involve a mechanism that revises Ig genes secondarily. PMID- 11410253 TI - Molecular dynamics of point mutated I-A(k) molecules expressed on lymphocytes. AB - We have recently reported the lateral and rotational diffusion parameters for I A(k) molecules expressing various cytoplasmic truncations (Int. Immunol. 12 (2000) 1319). We now describe the membrane dynamics of I-A(k) with various mutations in the presumed contact region between alphabeta-heterodimers in an (alphabeta)2 dimer of dimers structure. Such mutations are known to strongly affect the antigen presentation ability of these molecules (Int. Immunol. 10 (1998) 1237-1249) but cause relatively small changes in the molecular dynamics of I-A(k). Lateral diffusion coefficients of I-A(k) wild-type molecules and mutants obtained via fringe fluorescence photobleaching recovery (FPR) ranged from 1.1 to 2.3x10(-10)cm2/s at room temperature while fractional mobilities averaged 75+/ 6%. For all cell types examined, treatment with either hen egg lysozyme 46-61 peptide or db-cAMP reduced the I-A(k) mobile fraction by about 10% relative to untreated cells, suggesting that these treatments may increase lateral confinement of class II in lipid rafts or cytoskeletal interactions of the molecules. Wild-type I-A(k) and mutants capable of normal or partial antigen presentation exhibited, as a group, slightly longer rotational correlation times (RCT) at 4 degrees C than did mutants inactive in antigen presentation, 14+/-4 versus 10+/-1 micros, respectively. Moreover, peptide, cAMP and anti-CD40 mAb treatment all increased rotational correlation times for fully- and partially functional I-A(k) but not for non-functional molecules. For example, 16 h peptide treatment yielded average RCTs of 28+/-12 and 10+/-1 micros for the groups of functional and non-functional molecules, respectively. Such modulation of the dynamics of functional class II molecules is consistent with these treatments' stabilization of class II or induction of new gene expression. Measurements of fluorescence resonant energy transfer between I-A(k), though complicated by cellular autofluorescence, averaged 6+/-7% over 15 cells or treatments, a result consistent with the presence of a small fraction of I-A(k) as a dimer of dimers species. In summary, our results suggest subtle changes in the molecular motions of class II molecules correlate with a significant impact on class II function. Molecules active in antigen presentation exhibit more restricted motion in the membrane, and thus presumably more extensive intermolecular interactions, than non-functional molecules. Further, treatments, such as db-cAMP and anti-CD40, which rescue antigen presentation by partially defective mutants, appear to increase such interactions, several types of which have already been reported for class II. A more detailed understanding of these phenomena will require both more sensitive biophysical tools and a more refined model of the role of class II intermolecular interactions in antigen presentation. PMID- 11410254 TI - Neuroscience in the post-genome era: an overview. AB - Although sequencing of the human genome has been taking place in rapidly accelerating fashion for years, the presentation of the entire sequence (to a first approximation, anyway) has now permitted a global view of its structure. The completion of the sequence has therefore invited all manner of aesthetic, philosophical, societal - as well as biological - discussions of its implications. This global view of the genome will undoubtedly forever change the face of biology. But how? Here are a few perspectives from the point of view of neuroscience. PMID- 11410255 TI - Kainate receptors: knocking out plasticity. AB - There is increasing evidence that kainate receptors contribute to both postsynaptic and presynaptic signaling. Studies of knockout mice have played a pivotal role in defining the functions of kainate receptors, including a recent study that implicates kainate receptors in frequency-dependent facilitation and long-term potentiation of hippocampal mossy fiber synapses. PMID- 11410267 TI - Short-term synaptic plasticity as a temporal filter. AB - Synaptic efficacy can increase (synaptic facilitation) or decrease (synaptic depression) markedly within milliseconds after the onset of specific temporal patterns of activity. Recent evidence suggests that short-term synaptic depression contributes to low-pass temporal filtering, and can account for a well known paradox - many low-pass neurons respond vigorously to transients and the onsets of high temporal-frequency stimuli. The use of depression for low-pass filtering, however, is itself a paradox; depression induced by ongoing high temporal frequency stimuli could preclude desired responses to low-temporal frequency information. This problem can be circumvented, however, by activation of short-term synaptic facilitation that maintains responses to low-temporal frequency information. Such short-term plasticity might also contribute to spatio temporal processing. PMID- 11410269 TI - Brain damage, sex hormones and recovery: a new role for progesterone and estrogen? AB - Estrogen and progesterone, long considered for their roles as primary hormones in reproductive and maternal behavior, are now being studied as neuroprotective and neuroregenerative agents in stroke and traumatic brain injuries. Collectively, the hormones reduce the consequences of the injury cascade by enhancing anti oxidant mechanisms, reducing excitotoxicity (altering glutamate receptor activity, reducing immune inflammation, providing neurotrophic support, stimulating axonal remyelinization), and enhancing synaptogenesis and dendritic arborization. Estrogen seems more effective as a prophylactic treatment in females at risk for cardiac and ischemic brain injury, whereas progesterone appears to be more helpful in the post-injury treatment of both male and female subjects with acute traumatic brain damage. However, a recent clinical trial with estradiol replacement therapy in elderly women that have a history of cerebrovascular disease, showed that this hormone was unable to protect against reoccurrence of ischemia or to reduce the incidence of mortality compared to a placebo. PMID- 11410270 TI - Advances in molecular genetics and pathology of cerebrovascular disorders. AB - Progress in molecular genetics has enabled the dissection of several autosomal dominantly inherited forms of cerebrovascular disorders. Mutations in diverse genes might induce pathological changes in intracranial vessels, resulting in cerebral haemorrhages and ischaemic strokes. Such pathologies, however, might also result from systemic vascular disease caused by mutations or polymorphisms in genes that regulate cardiovascular physiology, blood coagulation, lipid metabolism and metabolic functions. Interestingly, several mutations that directly affect CNS vasculature involve genes that control inter- or intracellular signalling functions. Although highly variable phenotypes make it difficult to pinpoint the genotypes, genetic characterization of cerebrovascular disorders is valuable for understanding the pathogenesis and management of sporadic disease. PMID- 11410271 TI - The larval ascidian nervous system: the chordate brain from its small beginnings. AB - The body plan of the tadpole larva of ascidians, or sea-squirts, is widely presumed to be close to that of the hypothetical ancestor of all chordate animal groups, including vertebrates. This is nowhere more obvious than in the organization and development of the dorsal tubular nervous system. Several recent developments advocate this model neural system for studies on neurobiology and neurogenesis. These include advances in our understanding of development in ascidian embryos and of differentiation among the cellular progeny of its neural plate; the application of transgenic and mutant approaches to studies on ascidian larval neurones; and the prospect of advances in genomic analyses. In addition to providing ways to study a working chordate brain in miniature, all these offer insights into the ancestral condition of the developing vertebrate brain. PMID- 11410272 TI - Oxidative stress and neuroAIDS: triggers, modulators and novel antioxidants. AB - Neurological disorders represent one of the most common disturbances accompanying HIV infection. In the past few years, highly antiretroviral active therapy has significantly reduced the incidence of HIV-related diseases. However, neurological dysfunction in AIDS patients still remains an unresolved problem. Oxidative stress, which occurs in brain tissues of patients undergoing HIV infection and is implicated in cell death of both astroglia and neurones, has recently been suggested to play a role in the pathogenesis of neuroAIDS. Thus, a better understanding of the processes that trigger and modulate free radical formation in brain tissues of AIDS patients might help in a successful therapeutic approach to the neuropathogenesis of HIV infection. PMID- 11410273 TI - Intrinsic and extrinsic factors that shape neocortical specification. AB - Increasing evidence points to the importance of intrinsic molecular cues in specifying the regional identity of mammalian neocortex. Few such cues, however, have been found to be restricted to individual functionally defined cortical areas before the arrival of afferent information. In contrast, thalamocortical axons are specifically targeted to individual cortical areas, raising the possibility that they can instruct some aspects of cortical areal identity. Cortical structure and function can be altered by modifying the source or pattern of activity in thalamocortical afferents. In particular, studies of cross-modal plasticity have shown that in many respects, one sensory cortical area can substitute for another after a switch of input modality during development. Afferent inputs might therefore direct the formation of their own processing circuitry, a possibility that has important implications for brain development, plasticity and evolution. PMID- 11410274 TI - Structural features of a snake venom thrombin-like enzyme: thrombin and trypsin on a single catalytic platform? AB - The Lachesis muta thrombin-like enzyme (LM-TL) is a single chain serine protease that shares 38% sequence identity with the serine protease domain of thrombin and also displays similar fibrinogen-clotting activity. In addition, the 228 amino acid residue LM-TL is 52% identical to trypsin, and cleaves chromogenic substrates with similar specificity. Herein we report a three-dimensional (3D) model validated experimentally for LM-TL based on these two homologous proteins of known 3D structure. Spatial modeling of LM-TL reveals a serine protease with a chymotrypsin fold presenting a hydrophobic pocket on its surface, involved in substrate recognition, and an important 90's loop, involved in restricting the LM TL catalytic site cleft. Docking analysis showed that LM-TL would not form a stable complex with basic pancreatic trypsin inhibitor and wild-type ecotin since its 90's loop would restrict the access to the catalytic site. LM-TL formed acceptable interactions with fibrinopeptide A and a variant of ecotin; ecotin TSRR/R in which both the primary and secondary binding sites are mutated Val81Thr, Thr83Ser, Met84Arg, Met85Arg and Asp70Arg. Furthermore, analysis of the primary structures of LM-TL and of the seven snake venom thrombin-like enzymes (SVTLEs) family reveals a subgroup formed by LM-TL, crotalase, and bilineobin, both closely related to thrombin. Therefore, LM-TL provides an initial point to compare SVTLEs with their counterparts, e.g. the mammalian serine proteases, and a basis for the localization of important residues within the little known SVTLEs family. PMID- 11410275 TI - Inactivation of CMY-2 beta-lactamase by tazobactam: initial mass spectroscopic characterization. AB - The CMY-2 beta-lactamase, a plasmid determined class C cephalosporinase, was shown to be susceptible to inhibition by tazobactam (K(i)=40 microM). The reaction product(s) of CMY-2 beta-lactamase with the beta-lactamase inhibitor tazobactam were analyzed by electrospray ionization/mass spectrometry (ESI/MS) to characterize the prominent intermediates of the inactivation pathway. The ESI/MS determined mass of CMY-2 beta-lactamase was 39851+/-3 Da. After inactivating CMY 2 beta-lactamase with excess tazobactam, a single species, M(r)=39931+/-3.0, was detected. Comparison of the peptide maps from tryptic digestion of the native enzyme and the inactivated beta-lactamase followed by LC/MS identified two 22 amino acid peptides containing the active site Ser64 modified by a fragment of tazobactam. These two peptides were increased in mass by 70 and 88 Da, respectively. UV difference spectra following inactivation revealed the presence of a new species with a 302 nm lambda(max). Based upon the increase in molecular mass of the tazobactam inactivated CMY-2 beta-lactamase, we propose that during the inactivation of this beta-lactamase by tazobactam an imine is formed. Tautomerization forms the spectrally observed enamine. Hydrolysis generates the covalently attached malonyl semialdehyde, its hydrate, or an enol. This work provides information on the mass of a stable enzyme intermediate of a class C beta-lactamase inactivated by tazobactam and, for the first time, unequivocal evidence that a cross-linked species is not required for apparent inactivation. PMID- 11410276 TI - Inactivation of the serpin alpha(2)-antiplasmin by stromelysin-1. AB - Matrix metalloproteinase-3 (MMP-3 or stromelysin-1) hydrolyzes the Met(374) Ser(375) (P3-P2), Glu(416)-Leu(417) and Ser(432)-Leu(433) peptide bonds in human alpha(2)-antiplasmin (alpha(2)-AP), the main physiological plasmin inhibitor. Cleavage is completely abolished in the presence of the MMP inhibitors EDTA or 1,10-phenanthroline. At enzyme/substrate ratio of 1:10 at 37 degrees C, alpha(2) AP protein cleavage occurs with a half-life of 8 min, and is associated with rapid loss of inhibitory activity towards plasmin with a half-life of 5 min. alpha(2)-AP cleaved by MMP-3 does no longer form a stable complex with plasmin, as shown by SDS-PAGE, and does no longer interact with plasminogen, as shown by crossed immunoelectrophoresis with plasminogen added to the gel. These data are compatible with the removal of a COOH-terminal fragment containing the reactive site peptide bond and the plasmin(ogen)-binding site. In addition, MMP-3 cleaves the Pro(19)-Leu(20) peptide bond in alpha(2)-AP, thereby removing the fibrin binding site from the inhibitor. A dysfunctional alpha(2)-AP variant (Ala alpha(2)-AP or alpha(2)-AP Enschede), with an alanine insertion in the reactive site sequence converting it from a plasmin inhibitor into a substrate, was also efficiently cleaved by MMP-3 (half-life of 13 min at 37 degrees C and enzyme/substrate ratio of 1:10). Cleavage and inactivation of alpha(2)-AP by MMP 3 may constitute a mechanism favoring local plasmin-mediated proteolysis. PMID- 11410277 TI - Enhancement of the thermal stability of pyroglutamyl peptidase I by introduction of an intersubunit disulfide bond. AB - From the comparison of the three-dimensional structure of mesophilic pyroglutamyl peptidase from Bacillus amyloliquefaciens and the thermophilic enzyme from Thermococcus litoralis, the intersubunit disulfide bond was estimated to be one of the factors for thermal stability. Since Ser185 was corresponded to Cys190 of the thermophilic enzyme by sequence alignment, the Ser185 residue was replaced with cysteine by site-directed mutagenesis. The S185C mutant enzyme appeared to form a disulfide bond, which was confirmed by SDS-PAGE with and without 2 mercaptoethanol. The mutant enzyme showed a catalytic efficiency equivalent to that of the wild-type enzyme for hydrolysis of a synthetic peptide substrate. However, the thermal stability of the S185C mutant was found to be 30 degrees C higher than that of wild-type. Thus the introduction of a disulfide bond enhanced thermal stability without changing the catalytic efficiency of the enzyme. PMID- 11410278 TI - Comparison of the decameric structure of peroxiredoxin-II by transmission electron microscopy and X-ray crystallography. AB - The decameric human erythrocyte protein torin is identical to the thiol-specific antioxidant protein-II (TSA-II), also termed peroxiredoxin-II (Prx-II). Single particle analysis from electron micrographs of Prx-II molecules homogeneously orientated across holes in the presence of a thin film of ammonium molybdate and trehalose has facilitated the production of a >/=20 A 3-D reconstruction by angular reconstitution that emphasises the D5 symmetry of the ring-like decamer. The X-ray structure for Prx-II was fitted into the transmission electron microscopic reconstruction by molecular replacement. The surface-rendered transmission electron microscopy (TEM) reconstruction correlates well with the solvent-excluded surface of the X-ray structure of the Prx-II molecule. This provides confirmation that transmission electron microscopy of negatively stained specimens, despite limited resolution, has the potential to reveal a valid representation of surface features of protein molecules. 2-D crystallisation of the Prx-II protein on mica as part of a TEM study resulted in the formation of a p2 crystal form with parallel linear arrays of stacked rings. This latter 2-D form correlates well with that observed from the 2.7 A X-ray structure of Prx-II solved from a new orthorhombic 3-D crystal form. PMID- 11410279 TI - Isolation and characterization of type I antifreeze proteins from Atlantic snailfish (Liparis atlanticus) and dusky snailfish (Liparis gibbus). AB - Antifreeze proteins (AFPs) were isolated from the blood plasma of Atlantic snailfish Liparis atlanticus and dusky snailfish Liparis gibbus, which belong to the Teleost family Cyclopteridae, a close relative of sculpins. Using a combination of gel filtration chromatography and reversed-phase HPLC, proteins were purified to individual peaks. Atlantic snailfish plasma contained two different proteins (MW=9344, 9415) while dusky snailfish plasma contained five protein isoforms (MW=9514-9814), as determined by mass spectrometry. Further characterization revealed that these proteins are rich in alanine (>50 mol%), and have alpha-helical secondary structure that can undergo reversible thermal denaturation. Thermal hysteresis activities of these proteins were similar to each other but lower than the major type I AFPs from winter flounder. Results of this study have indicated that although the AFPs from snailfish are significantly larger than previously described type I AFPs, they share enough characteristics to be classified in this group. PMID- 11410280 TI - The mechanism of the phosphoryl transfer catalyzed by Yersinia protein-tyrosine phosphatase: a computational and isotope effect study. AB - In order to evaluate various mechanistic proposals that have been made regarding the mechanism of the first step of the reaction catalyzed by protein-tyrosine phosphatases, new experimental data have been obtained, and some existing data have been carefully reevaluated. New kinetic isotope effect data for the uncatalyzed hydrolysis of p-nitrophenyl phosphate allow a better evaluation of previously reported data from enzymatic reactions with this substrate. The interpretation, and misinterpretation, of pH rate studies is considered. The pathway of this reaction has been modeled computationally and is found to be generally consistent with experimental studies, except for the extent of proton transfer to the leaving group. PMID- 11410281 TI - Mammalian intestinal alkaline phosphatase acts as highly active exopolyphosphatase. AB - Recent results revealed that inorganic polyphosphates (polyP), being energy-rich linear polymers of orthophosphate residues known from bacteria and yeast, also exist in higher eukaryotes. However, the enzymatic basis of their metabolism especially in mammalian cells is still uncertain. Here we demonstrate for the first time that alkaline phosphatase from calf intestine (CIAP) is able to cleave polyP molecules up to a chain length of about 800. The enzyme acts as an exopolyphosphatase degrading polyP in a processive manner. The pH optimum is in the alkaline range. Divalent cations are not required for catalytic activity but inhibit the degradation of polyP. The rate of hydrolysis of short-chain polyP by CIAP is comparable to that of the standard alkaline phosphatase (AP) substrate p nitrophenyl phosphate. The specific activity of the enzyme decreases with increasing chain length of the polymer both in the alkaline and in the neutral pH range. The K(m) of the enzyme also decreases with increasing chain length. The mammalian tissue non-specific isoform of AP was not able to hydrolyze polyP under the conditions applied while the placental-type AP and the bacterial (Escherichia coli) AP displayed polyP-degrading activity. PMID- 11410282 TI - Lipase-catalyzed synthesis of isoamyl butyrate. A kinetic study. AB - Kinetics of lipase-catalyzed esterification of butyric acid and isoamyl alcohol have been investigated. The reaction rate could be described in terms of the Michaelis-Menten equation with a Ping-Pong Bi-Bi mechanism and competitive inhibition by both the substrates. No evidence of any significant diffusional limitations was detected that could affect the kinetics. The values of the apparent kinetic parameters were computed as: V(max)=11.72 micromol/min/mg; K(M, Acid)=0.00303 M; K(M, Alcohol)=0.00306 M; K(i, Acid)=1.05 M; and K(i, Alcohol)=6.55 M. This study indicates a competitive enzyme inhibition by butyric acid during lipase-catalyzed esterification reaction. Butyric acid, being a short chain polar acid, concentrates in the microaqueous layer and causes a pH drop in the enzyme microenvironment leading to enzyme inactivation. Butyric acid binds to acyl-enzyme complex unproductively to yield a dead-end intermediate that can no longer give rise to an ester. High concentration of butyric acid gave rise to inactivation of the biocatalyst in addition to dead-end inhibition. PMID- 11410283 TI - Investigation of human hair cuticle structure by microdiffraction: direct observation of cell membrane complex swelling. AB - The cuticle of mammalian hair fibres protects the core of the fibre against physical and chemical stress. The structure and some of the properties of the cuticle have been extensively studied by electron microscopy. However, there is still a need for a less invasive structural probe. For this purpose, microdiffraction experiments have been carried out on human hair samples showing a characteristic small-angle X-ray scattering pattern for the cuticle. This pattern has been assigned to the cell membrane complex (CMC) between each cuticle scale. Using a simple model of the electron density within the CMC, values have been derived for the average thickness of the beta- and delta-layers which are close to those obtained by electron microscopy. In order to illustrate the potentialities of microdiffraction in studying the properties of the cuticle, the effect of water sorption has been monitored. Using the intensity modelling described above, a 10% swelling of the delta-layer's thickness has been observed. This study shows that structural modifications of the CMC by physical or chemical stress can be followed directly on the cuticle of human hair fibres by microdiffraction analysis. PMID- 11410284 TI - Molecular, biochemical and immunological analyses of porcine pancreatic DNase I. AB - Deoxyribonuclease I (DNase I) was purified 26500-fold in 39% yield from porcine pancreas to electrophoretic homogeneity using three-step column chromatography. The purified enzyme was inhibited by an antibody specific to the purified enzyme but not by G-actin. A 1303 bp cDNA encoding porcine DNase I was constructed from total RNA from porcine small intestine using a rapid amplification of cDNA ends method, followed by sequencing. Mature porcine DNase I protein was found to consist of 262 amino acids. Unlike all other mammalian DNase I enzymes that are inhibited by G-actin, porcine DNase I has H65 and S114 instead of Y65 and A114, which presumably results in the lack of inhibition. Porcine DNase I was more sensitive to low pH than rat or bovine enzymes. Compared with their primary structures, the amino acid at position 110 was N in porcine enzyme, but S in rat and bovine enzymes. A porcine mutant enzyme in which N was substituted by S alone at position 110 (N110S) became resistant to low pH to a similar extent as the rat and bovine enzymes. PMID- 11410285 TI - Purification and catalytic properties of the chlorophenol 4-monooxygenase from Burkholderia cepacia strain AC1100. AB - Burkholderia cepacia strain AC1100 can be induced for the degradation of 2,4,5 trichlorophenol (2,4,5-TCP). We have purified the active enzyme 30-fold to apparent homogeneity with a 44% yield by a two-step chromatographic procedure, and showed that it consists of a single type of subunit of 59 kDa based on SDS PAGE using Coomassie blue and Sypro staining. This enzyme has no bound prosthetic group but requires exogenous addition of FAD and NADH to perform the dioxygen dependent hydroxylation in the 4-position of 2,4,6-TCP. Studies of the stoichiometry revealed the consumption of 2 mol of NADH plus 1 mol of dioxygen per mol of 2,4,6-TCP with identification of the reaction product as 2,6 dichlorohydroquinone. Steady state kinetic parameters for cofactors and a variety of substrates were determined. Low K(m) values of 1+/-0.1 microM, 32+/-5 microM and 4+/-2 microM were found for FAD, NADH and 2,6-dichlorophenol (2,6-DCP), respectively, under saturating conditions for the two others. In the presence of 2,6-DCP as a substrate, methimazole (MMI) inhibited the enzyme competitively with a K(i)=27 microM. When other polychlorinated substrates were studied, IC(50) values for MMI were found in a range compatible with their apparent affinity. On the basis of aromatic product formation, NADH and O(2) consumption schemes for 2,4,6-TCP and 2,4,5-TCP degradation are discussed. A Blast search revealed that this enzyme has a high sequence identity (60%) with 2,4,6-TCP-4-monooxygenases from Burkholderia pickettii and from Azotobacter sp. strain GP1 which all of them catalyze para hydroxylative dehalogenation. PMID- 11410286 TI - Enzymatic properties of human hemalbumin. AB - The binding of hemin to the primary site of human serum albumin (HSA) has been reinvestigated using UV-Vis, CD and NMR techniques. The major fraction of bound hemin contains a five-coordinated high-spin iron(III) center, but a minor fraction of the metal appears to be in a six-coordinated, low-spin state, where a 'distal' residue, possibly a second histidine residue, completes the coordination sphere. The reduced, iron(II) form of the adduct contains six-coordinated low spin heme. The distal residue hinders the access to the iron(III) center of hemin HSA to small anionic ligands like azide and cyanide and destabilizes the binding of neutral diatomics like dioxygen and carbon monoxide to the iron(II) form. In spite of these limitations, the hemin-HSA complex promotes hydrogen peroxide activation processes that bear the characteristics of enzymatic reactions and may have biological relevance. The complex is in fact capable of catalyzing peroxidative reactions on phenolic compounds related to tyrosine and hydrogen peroxide dismutation. Kinetic and mechanistic studies confirm that the low efficiency with which peroxidative processes occur depends on the limited rate of the reaction between hydrogen peroxide and the iron(III) center, to form the active species, and by the competitive peroxide degradation reaction. PMID- 11410287 TI - The proapoptotic protein Bad binds the amphipathic groove of 14-3-3zeta. AB - Through interaction with a multitude of target proteins, 14-3-3 proteins participate in the regulation of diverse cellular processes including apoptosis. These 14-3-3-interacting proteins include a proapoptotic Bcl-2 homolog, Bad (Bcl 2/Bcl-XL-associated death promoter). To understand how 14-3-3 interacts with Bad and modulates its function, we have identified structural elements of 14-3-3 necessary for 14-3-3/Bad association. 14-3-3 contains a conserved amphipathic groove that is required for binding to several of its ligands. We used peptides of known binding specificity as competitors to demonstrate that Bad interacts with 14-3-3zeta via its amphipathic groove. More detailed analysis revealed that several conserved residues in the groove, including Lys-49, Val-176, and Leu-220, were critical for Bad interaction. These results were applied to investigations of the ability of 14-3-3 to prevent Bad-induced cell death. When co-expressed with Akt, wild-type 14-3-3 could reduce the ability of Bad to cause death, however 14-3-3zetaK49E, which cannot bind Bad, failed to inhibit Bad. It seems that the amphipathic groove of 14-3-3 represents a general binding site for multiple ligands, raising issues related to competition of ligands for 14-3-3. PMID- 11410288 TI - Use of silicate sol-gels to trap the R and T quaternary conformational states of pig kidney fructose-1,6-bisphosphatase. AB - Encapsulation of the homotetrameric pig kidney fructose-1,6-bisphosphatase (FBPase) in tetramethyl orthosilicate sol-gels was used to dramatically reduce the rate of the allosteric transition of the enzyme between the T and R allosteric states. When assayed in the absence of the allosteric inhibitor AMP, the enzyme encapsulated in the T-state exhibited little activity. The enzyme encapsulated in the R-state exhibited a 4-fold lower k(cat) and V(max) than the enzyme in solution, and the apparent K(m) for this enzyme was 350-fold higher than the corresponding value for the enzyme in solution. The [Mg(2+)](0.5) for the encapsulated enzyme was only 0.1 mM, compared to 0.54 mM for the normal enzyme. Magnesium activation, under both sets of conditions, was cooperative with a Hill coefficient of approximately 2. The activity of enzyme encapsulated in the R-state decreased to about 70% of initial activity within 1 min of adding AMP, it then decreased slowly to about 40% of initial activity over the following 7 h. Under the conditions tested, the encapsulated enzyme never became completely inactivated and AMP inhibition was no longer cooperative. For enzyme encapsulated in the T-state, activity was restored over approximately 7 h after removal of the AMP. The biphasic and slow responses to changing AMP levels suggest that encapsulated enzyme can be used to study the effects of local conformational changes distinct from the global quaternary conformational changes by slowing down the ability of the enzyme to carry out global rotations. The response to AMP exhibited by the encapsulated enzyme is consistent with the ability of AMP, at least partially, to directly influence the activity of the active site within each subunit. PMID- 11410289 TI - Impact of the tryptophan residues of Humicola lanuginosa lipase on its thermal stability. AB - Thermal stability of wild type Humicola lanuginosa lipase (wt HLL) and its two mutants, W89L and the single Trp mutant W89m (W117F, W221H, and W260H), were compared. Differential scanning calorimetry revealed unfolding of HLL at T(d)=74.4 degrees C whereas for W89L and W89m this endotherm was decreased to 68.6 and 62 degrees C, respectively, demonstrating significant contribution of the above Trp residues to the structural stability of HLL. Fluorescence emission spectra revealed the average microenvironment of Trps of wt HLL and W89L to become more hydrophilic at elevated temperatures whereas the opposite was true for W89m. These changes in steady-state emission were sharp, with midpoints (T(m)) at approx. 70.5, 61.0, and 65.5 degrees C for wt HLL, W89L, and W89m, respectively. Both steady-state and time resolved fluorescence spectroscopy further indicated that upon increasing temperature, the local movements of tryptophan(s) in these lipases were first attenuated. However, faster mobilities became evident when the unfolding temperatures (T(m)) were exceeded, and the lipases became less compact as indicated by the increased hydrodynamic radii. Even at high temperatures (up to 85 degrees C) a significant extent of tertiary and secondary structure was revealed by circular dichroism. Activity measurements are in agreement with increased amplitudes of conformational fluctuations of HLL with temperature. Our results also indicate that the thermal unfolding of these lipases is not a two-state process but involves intermediate states. Interestingly, a heating and cooling cycle enhanced the activity of the lipases, suggesting the protein to be trapped in an intermediate, higher energy state. The present data show that the mutations, especially W89L in the lid, contribute significantly to the stability, structure and activity of HLL. PMID- 11410290 TI - Formation of thermally induced aggregates of the soya globulin beta-conglycinin. AB - The effect of ionic strength (I) on the formation of thermally induced aggregates by the 7S globular storage protein of soya, beta-conglycinin, has been studied using atomic force microscopy. Aggregates were only apparent when I> or =0.1, and had a fibrous appearance, with a height (diameter) of 8-11 nm. At high ionic strength (I=1.0) the aggregates appeared to associate into clumps. When aggregate formation was studied at I=0.2, it was clear that aggregation only began at temperatures above the main thermal transition for the protein at 75 degrees C, as determined by differential scanning calorimetry. This coincided with a small change in secondary structure, as indicated by circular dichroism spectroscopy, suggesting that a degree of unfolding was necessary for aggregation to proceed. Despite prolonged heating the size of the aggregates did not increase indefinitely, suggesting that certain beta-conglycinin isoforms were able to act as chain terminators. At higher protein concentrations (1% w/v) the linear aggregates appeared to form large macroaggregates, which may be the precursors of protein gel formation. The ability of beta-conglycinin to form such distinctive aggregates is discussed in relation to the presence of acidic inserts in certain of the beta-conglycinin subunits, which may play an important role in limiting aggregate length. PMID- 11410291 TI - Characterization of the oxidative 3alpha-hydroxysteroid dehydrogenase activity of human recombinant 11-cis-retinol dehydrogenase. AB - 11-cis-Retinol dehydrogenase catalyzes the oxidation of cis-retinols, a rate limiting step in the biosynthesis of 9-cis-retinoic acid. It is also active toward 3alpha-hydroxysteroids, and thus might be involved in steroid metabolism. To better understand the role of this enzyme, we produced stable transfectants expressing 11-cis-retinol dehydrogenase in human embryonic kidney 293 cells. In vitro enzymatic assays have demonstrated that, with an appropriate exogenous cofactor, the enzyme catalyzes the interconversion of 5alpha-androstane 3alpha,17beta-diol and dihydrotestosterone and that of androsterone and androstanedione. However, using intact transfected cells, we found that the enzyme catalyzes reactions only in the oxidative direction. Thus, it is possible that 5alpha-androstane-3alpha,17beta-diol (an inactive androgen) can be converted into dihydrotestosterone, the most potent androgen, by the action of 11-cis retinol dehydrogenase. This reaction could constitute a non-classical pathway of production of active androgens in the peripheral tissues. We also showed that all trans-, 9-cis- and 13-cis-retinol inhibit the oxidative 3alpha-hydroxysteroid steroid activity of 11-cis-retinol dehydrogenase with similar K(i) values. Since all-trans-retinol is a precursor of cis-retinols, its inhibitory effect on the activity suggests that it could play an important role in modulating the formation of 9-cis-retinoic acid. In addition, we examined the effect of several known enzyme modulators, namely carbenoxolone, phenylarsine oxide and phosphatidylcholine, on 11-cis-retinol dehydrogenase activity. Taken together, our results suggest that, in humans, this enzyme might play a role in the biosynthesis of both 9-cis-retinoic acid and dihydrotestosterone. PMID- 11410292 TI - Calorimetric and Fourier transform infrared spectroscopic study of solid proteins immersed in low water organic solvents. AB - Calorimetric heat effects and structural rearrangements assessed by means of Fourier transform infrared (FTIR) amide I spectra were followed by immersing dry human serum albumin and bovine pancreatic alpha-chymotrypsin in low water organic solvents and in pure water at 298 K. Enthalpy changes upon immersion of the proteins in different media are in a good linear correlation with the corresponding IR absorbance changes. Based on calorimetric and FTIR data the solvents were divided into two groups. The first group includes carbon tetrachloride, benzene, nitromethane, acetonitrile, 1,4-dioxane, n-butanol, n propanol and pyridine where no significant heat evolution and structural changes were found during protein immersion. Due to kinetic reasons no significant protein-solvent interactions are expected in such systems. The second group of solvents includes dimethyl sulfoxide, methanol, ethanol, and water. Immersion of proteins in these media results in protein swelling and involves significant exothermic heat evolution and structural changes in the protein. Dividing of different media in the two groups is in a qualitative correlation with the solvent hydrophilicity defined as partial excess molar Gibbs free energy of water at infinite dilution in a given solvent. The first group includes the solvents with hydrophilicity exceeding 2.7 kJ/mol. More hydrophilic second group solvents have this energy values less than 2.3 kJ/mol. The hydrogen bond donating ability of the solvents also assists in protein swelling. Hydrogen bonding between protein and solvent is assumed to be a main factor controlling the swelling of dry solid proteins in the studied solvents. PMID- 11410293 TI - Enhancement of stability and activity of phospholipase A(1) in organic solvents by directed evolution. AB - We attempted to apply the directed evolution approach to enhancing enzyme properties in the presence of organic solvents, in which enzyme stability and activity were often drastically reduced. Stability and catalytic activity of phospholipase A(1) in the presence of an organic solvent were enhanced by error prone polymerase chain reaction (PCR) and DNA shuffling followed by a filter based visual screening. Three mutants (SA8, SA17 and SA20) were isolated on indicator plates (i.e., 1% phosphatidylcholine gels containing 30% dimethyl sulfoxide (DMSO)) after a second mutant library was treated in 50% DMSO for 36 h. The half-life values of the three mutants exhibited an approximately 4-fold increase. The three mutants also exhibited increased stability in all organic solvents tested compared with the wild-type enzyme. Thus, an enzyme variant having superior catalytic efficiency in most of the organic solvents could be obtained by using any solvent suitable for designing the efficient screening system, regardless of the properties of the particular solvent. PMID- 11410294 TI - Conjugation of phenylalanine hydroxylase with polyubiquitin chains catalysed by rat liver enzymes. AB - Phenylalanine hydroxylase (PAH, EC 1.14.16.1) is a highly regulated liver enzyme which catalyses the conversion of L-phenylalanine to L-tyrosine, the rate limiting step in the catabolic pathway of this amino acid. Among the approx. 400 different mutations of human (h) PAH, frequently associated with the metabolic disease phenylketonuria, a low stability is a characteristic property when expressed in eucaryotic cells. In this study, the pathway of hPAH degradation is addressed with focus on its conjugation with polyubiquitin chains catalysed by the ubiquitin-conjugating enzyme system (E1, E2, E3) isolated from rat liver by covalent affinity chromatography on ubiquitin-Sepharose. In the reconstituted in vitro ubiquitination assay, the enzyme system catalysed both the formation of free polyubiquitin chains and the polyubiquitination of wild-type (wt) hPAH and its 'catalytic domain' (DeltaN102/DeltaC24-hPAH) as visualized by two-dimensional electrophoresis. The ubiquitination of wt-PAH may play a role in the degradation of this liver enzyme notably of its many unstable disease-associated mutant forms. The present approach may also have a more general application in the study of liver proteins as possible targets for ubiquitination. PMID- 11410295 TI - Purification and characterization of active recombinant rat kallikrein rK9. AB - The rat tissue kallikrein rK9 is most abundant in the submandibular gland and the prostate. It has been successfully expressed in the Pichia pastoris yeast expression system. A full-length cDNA coding for the mature rK9 was fused in frame with yeast alpha-factor cDNA. The fusion protein was secreted into the medium with high yield without being processed by the yeast KEX2 signal peptidase. Mature rK9 was efficiently released from the fusion protein by trypsin and was purified to homogeneity by one-step affinity chromatography using soya bean trypsin inhibitor (SBTI) as affinity ligand. The identity of the recombinant enzyme was checked by N-terminal amino acid sequencing, Western blot analysis and kinetic studies. The dual trypsin- and chymotrypsin-like enzymatic specificity of rK9 was assessed by determining specificity constants (k(cat)/K(m)) for the hydrolysis of fluorogenic substrates, the peptide sequences of which were derived from proparathyroid hormone (pro-PTH) and from semenogelin-I. Our results confirmed the presence of an extended binding site in the rK9 active site. We also identified a far more sensitive substrate of this enzyme than those previously described, Abz-VKKRSARQ-EDDnp, which was hydrolysed with a catalytic efficiency k(cat)/K(m) of 420000 M(-1)s(-1). Finally, we showed that four of the five major proteins contained in secretions of rat seminal vesicles were rapidly degraded by recombinant rK9. PMID- 11410296 TI - Temperature-induced changes in copper centers and protein conformation of two fungal laccases from Coriolus hirsutus and Coriolus zonatus. AB - The paper reports on two fungal laccases from Coriolus hirsutus and Coriolus zonatus and their type-2 copper-depleted derivatives. Temperature-induced changes of the copper centers were characterized by optical and electron paramagnetic resonance (EPR) spectroscopy, and the overall protein stability by differential scanning microcalorimetry. The intact enzymes showed highly cooperative thermal unfolding transitions at about 90 degrees C. Type-2 copper depletion led to uncoupling of the domains characterized by a different melting pattern which resolved three subtransitions. Melting curves monitored optically at 290, 340 and 610 nm showed additional transitions below thermal unfolding temperature. EPR spectra of the intact laccases showed the disintegration of the trinuclear copper cluster accompanied by loss of one of the copper ions and disappearance of the strong antiferromagnetic coupling in the type-3 site at 70 degrees C and above 70 degrees C. The copper centers of type-2 copper-depleted laccase showed reduced thermotolerance. PMID- 11410297 TI - Utilization of peroxide and its relevance in oxygen insertion reactions catalyzed by chloroperoxidase. AB - Chloroperoxidase (CPO) catalyzed oxygen insertions are highly enantioselective and hence of immense biotechnological potential. A peroxide activation step is required to give rise to the compound I species that catalyzes this chiral reaction. A side reaction, a catalase type peroxide dismutation, is another feature of CPO's versatility. This work systematically investigates the utilization of different peroxides for the two reactions, i.e. the catalase type reaction and the oxygen insertion reaction. For the oxygen insertion reaction, indene and phenylethyl sulfide were chosen as substrate models for epoxidation and sulfoxidation respectively. The results clearly show that CPO is stable towards hydrogen peroxide and has a total number of turnovers near one million prior to deactivation. The epoxidation reactions terminate before completion because the enzyme functioning in its catalatic mode quickly removes all of the hydrogen peroxide from the reaction mixture. Sulfoxidation reactions are much faster than epoxidation reactions and thus are better able to compete with the catalase reaction for hydrogen peroxide utilization. A preliminary study towards optimizing the reaction system components for a laboratory scale synthetic epoxidation is reported. PMID- 11410298 TI - Radiation injury of the rectum. PMID- 11410299 TI - Variations of intracavitary applicator geometry during multiple HDR brachytherapy insertions in carcinoma cervix and its influence on reporting as per ICRU report 38. AB - PURPOSE: This paper examines the extent of variation in the applicator geometry during multiple high dose rate (HDR) intracavitary brachytherapy (ICBT) applications and its impact on reporting as per ICRU report 38. MATERIALS AND METHODS: Eighty orthogonal radiographs from 20 consecutive patients of carcinoma cervix (FIGO stages, IIA-IIIB) having four HDR ICBT applications of 6 Gy each at weekly intervals following teletherapy were evaluated. The applicator consisted of a flexible intrauterine tandem (IUT) independent of the ovoid assembly. The applicator geometry was evaluated in terms of: alpha angle, beta angle, intrauterine length (IUTL), interovoid (IOV), os to right ovoid (ORT) and os to left ovoid (OLT) distances along with vertical (VDL) and anteroposterior displacements (ADL) of the os with respect to the ovoids. The Cartesian co ordinates (X, Y, and Z) of the IUT tip, centre of both ovoids and os were also measured. Doses to right point A (ARD), left point A (ALD), along with a reference volume of 6 Gy for ICRU height (IRH), width (IRW), thickness (IRT) and volume (IRV) were estimated for each application. RESULTS: Highly significant differences (P<0.001) between four insertions in any given patient across 20 patients for alpha angle, beta angle, IUTL, IOV, ORT, VDL, co-ordinates of the IUT, ovoids and os were observed, except for ADL (P=0.041) and OLT (P=0.247). As a consequence, variations were observed in ARD (P=0.027), ALD (P=0.017); IRH, IRW, IRT and IRV (all P<0.001). Applicator factors which influenced the various dose specification parameters were: beta angle and ORT for both ARD and ALD; UTLN, VDL and ORT for IRH; UTLN and IOV for IRW; UTLN for IRT and VDL for the 6 Gy IRV. CONCLUSIONS: A significant variation of the applicator geometry and its movement was observed in patients undergoing multiple HDR ICBT. This could have implications for reporting dose and volume specifications as required by ICRU report 38. PMID- 11410300 TI - Set-up verification of cervix cancer patients treated with long treatment fields; implications of a non-rigid bony anatomy. AB - BACKGROUND AND PURPOSE: For cervix cancer patients, treatment fields may extend up to vertebra L1. In clinical practice, set-up verification is based on measured displacements of the pelvic rim as visible in the caudal part of the treatment fields. The implications of this procedure for the positions of bony structures in the cranial part of the fields were investigated. MATERIALS AND METHODS: Twelve patients had four repeat simulator sessions. Both during treatment simulation (the reference) and the repeat sessions, anterior radiographs were acquired covering the whole treatment field. The films were used to investigate differences between the cranial and the caudal parts of the treatment field in day-to-day bony anatomy displacements. RESULTS: Both in the transversal and the longitudinal directions, these differences were significant (3.5 mm, 1 SD). Indications were found that large differences in the cranio-caudal direction may be correlated with (non-rigid) internal pelvic rim rotations around a lateral axis. In the longitudinal direction, the position of L1 correlated much better with the position of vertebra S1 than with the position of the pelvic rim, which is usually used for set-up verification. CONCLUSIONS: Due to the non-rigid bony anatomy of the studied patients, the usual set-up verification and correction procedure can result in set-up errors of 10 mm and more for structures in the cranial part of the treatment field, even in the case of a perfect set-up of the pelvic rim. Possibly, other patient set-up and immobilization procedures may result in a better day-to-day reproducibility of the 3D bony anatomy shape. (Remaining) Differences in anatomy position changes between the caudal and cranial field ends may be accounted for by using non-uniform clinical target volume-to-planning target volume margins, or by an adapted patient set-up verification and correction protocol. PMID- 11410301 TI - Three-dimensional computed tomography-guided monotherapeutic pararectal brachytherapy of prostate cancer with seminal vesicle invasion. AB - PURPOSE: To treat patients with prostate cancer and seminal vesicle invasion with monotherapeutic three dimensional computed tomography (3-DCT)-guided posterior pararectal brachytherapy. METHODS AND MATERIALS: Three hundred and sixty two patients with clinical stage T1 a,b or T2 a,b of prostate cancer were referred for 3-DCT-guided brachytherapy. Each underwent ftirther staging with 3-D CT guided pararectal biopsy of the seminal vesicles under local anesthesia during the pre-treatment CT-planning. Forty-three patients (12%) were upstaged to T3 cNoMo disease. In the set of 43 patients, Eight had Gleason's score< or =6, 24 Gleason's score=7, and 11 patients > or =8. Initial PSA was <10 ng/ml in 14 patients, 10-20 ng/ml in 11 patients, and >20 in 18 patients. Of the 43 patients, 37 patients were treated monotherapeutically with 3-D CT-guided brachytherapy. No patients received hormone therapy after the implant. The prescribed dosage to the seminal vesicles and prostate is 120 Gy with Pd-103 seeds and 144 Gy with 1-125 seeds. RESULTS: The prescribed dosage was achieved in all 37 patient's throughout the seminal vesicles whose range of target radiation extended 5-10 mm outside the target in the adjacent fat as calculated with post-implant CT-dosimetry with Varian Brachy Vision or MMS software. Prostate Specific Antigen (PSA) outcome data were available in 34 patients treated with monotherapy and follow up ranged from 12-56 months (median, 24 months). Decreased PSA levels were stratified into six groups based on the presenting Gleason's score and initial PSA. In the first group (with Gleason's score< or =6 and initial PSA <20 ng/ml), PSA levels decreased to less than 0.5 ng/ml in all seven patients (100%) after brachytherapy. In the second group (with Gleason's=7 and initial PSA<20 ng/ml), PSA levels decreased to less than 1 ng/ml in 11 of 13 patients (85%); additionally PSA levels decreased to less than 0.5 ng/ml in ten patients (77% in this group). In the third group (with Gleason's score=7 and initial PSA> 20 ng/ml), PSA decreased to less than 0.5 ng/ml in four out of eight patients (50%). All of the patients in the fourth group (with Gleason's score> or =8 and initial PSA<20 ng/ml) decreased their PSA levels to less than 0.5 ng/ml in three of three patients. PSA decreased less than 0.5 ng/ml in two out of three patients (67% in the last group with Gleason's score> or =8 and initial PSA> 20 ng/ml). There were no patients with Gleason's score of 1-6 and greater than 20 ng/ml initial PSA. Patients, irrespective of the Gleason's score and PSA, had an overall response of decreased PSA (less than 1 ng/ml) of 79%. CONCLUSION: 3-D CT-guided brachytherapy delivers adequate dosage to the seminal vesicles. Clinical and biochemical results are encouraging in patients with low initial PSA levels regardless of their Gleason's scores, but longer-term data in a greater number of patients is necessary. PMID- 11410302 TI - The design and evaluation of a phantom for the audit of the treatment chain for prostate radiotherapy. AB - BACKGROUND AND PURPOSE: A phantom has been designed and built for a multi institutional technique audit of the planning and delivery for radiotherapy to the prostate. The phantom was designed to test both the geometric and dosimetric accuracy of each aspect of the process. MATERIALS AND METHODS: The phantom consists of two curved water filled perspex tanks either side of a central block of solid water equivalent material. There are two options for the central section; a target defining block and a dose measurement block. The target defining block uses air holes to define a 3-D target volume for imaging via a CT scanner or a simulator. These holes can subsequently be filled with steel pins to allow megavoltage imaging. The dose measurement block allows thimble chamber measurements to be made at pre-selected points in a 5x5mm array. Five dose measurement points, typical for a prostate planning target volume (PTV) were selected. Initial evaluation of the phantom was performed by auditing the prostate radiotherapy planning and treatment chain at one institution. RESULTS: Agreement between the phantom and planned geometry confirmed that the stages of image acquisition, transfer and manipulation were accurately performed. Agreement within 0.5% was found between phantom and water tank measurements for dose calibration at a reference point. The measured dose delivered was within 2% of the dose calculated by the planning computer for all of the selected measurement points. The target volume marked by the steel pins was visible using electronic portal imaging. CONCLUSIONS: The phantom is a useful tool for the technique audit of prostate radiotherapy. PMID- 11410303 TI - Annual rate of hearing loss falls after fractionated stereotactic irradiation for vestibular schwannoma. AB - PURPOSE: The rate of hearing loss in a population before and after irradiation was investigated to determine the effect of irradiation on hearing impairment. METHODS AND MATERIALS: In 72 patients with vestibular schwannoma who received fractionated stereotactic irradiation from 1992 to 1999, 21 had had their hearing levels examined 3 months or more before the treatment. The mean time between the initial examination and treatment was 18.6 months (range: 3-89 months), and the mean time between treatment and the last follow-up was 24.2 months (12-69 months). Thirty-six to 50 Gy in 20-25 fractions over 5 to 6 weeks was given using an X-ray beam from a linear accelerator. Pure tone average (PTA) was measured using the mean hearing level at five frequencies, and the annual rate of hearing loss was defined as [(hearing loss in PTA(dB))/(follow-up period (months)x12)]. RESULTS: The actual cumulative curve of decrease in tumor size of 2 mm or more was 38.3% at 2 years and 80.0% at 3 years. The mean of hearing loss in PTA was 11.6+/-10.3 dB (-1 to 35 dB) from the initial examination to the start of irradiation and 11.9+/-14.4 dB (-14 to 37 dB) from the start of irradiation to the last follow-up. The mean annual rates of hearing loss before irradiation and in the 1st,2nd,3rd and 4th years after irradiation were, respectively, 18.6, 11.2, 6.2, 5.1, and 5.0 dB/year. The annual rates of hearing loss in the 2nd year (P=0.025) and 3rd year (P=0.018) were significantly slower than the rate before irradiation. CONCLUSIONS: The mean annual rate of hearing loss was higher before irradiation than after irradiation, and hearing loss slowed rather than accelerated after irradiation. Although hearing loss after the treatment was usually permanent, fractionated stereotactic irradiation was suggested to be effective to lower the rate of hearing loss. PMID- 11410304 TI - Interobserver variations in gross tumor volume delineation of brain tumors on computed tomography and impact of magnetic resonance imaging. AB - PURPOSE: (1) To assess the interobserver variability of brain tumor delineation on computed tomography (CT). (2) To assess the impact of the addition of magnetic resonance imaging (MRI) information. METHODS: Nine physicians were asked to delineate the gross tumor volume (GTV) of five patients with supratentorial inoperable brain tumors on CT scans and 2 weeks (or more) later on MRIs. The delineations were performed on a computer screen. During delineation on MRI, the registered CT images (without delineation) were displayed on the screen (MRI+CT). RESULTS: A high interobserver variability in GTV delineation on CT is found: the ratio of the largest to the smallest defined volumes varies for the five patients by factors of resp. 2.8, 1.8, 1.8, 1.9 and 1.7. The interobserver variability is as large on MRI+CT as on CT alone (ratio largest/smallest volume: 2.4, 1.7, 1.9, 2.7 and 1.5). Volumes delineated on MRI+CT (mean: 69.6 cm(3)) are larger than on CT alone (mean: 59.5 cm(3)). Residual volumes (volume delineated on one image modality but not on the other) are >0 for CT alone and for MRI+CT. CONCLUSIONS: A large interobserver variability in GTV delineation of brain tumors is demonstrated. The addition of MRI to CT does not reduce interobserver variability. GTVs delineated on MRI+CT are larger than on CT alone, but some volumes are delineated on CT and not on MRI. Therefore, a combination of the two image modalities is recommended for brain tumor delineation for treatment planning. PMID- 11410305 TI - Radiosurgery for re-irradiation of brain metastasis: results in 54 patients. AB - PURPOSE: To evaluate in terms of probabilities of local-regional control and survival, as well as of treatment-related toxicity, results of radiosurgery for brain metastasis arising in previously irradiated territory. PATIENTS AND METHODS: Between January 1994 and March 2000, 54 consecutive patients presenting with 97 metastases relapsing after whole brain radiotherapy (WBRT) were treated with stereotactic radiotherapy. Median interval between the end of WBRT and radiosurgery was 9 months (range 2-70). Median age was 53 years (24-80), and median Karnofski performance status (KPS) 70 (60-100). Forty-seven patients had one radiosurgery, five had two and two had three. Median metastasis diameter and volume were 21 mm (6-59) and 1.2 cc (0.1-95.2), respectively. A Leksell stereotactic head frame (Leksell Model G, Elektra, Instrument, Tucker, GA) was applied under local anesthesia. Irradiation was delivered by a gantry mounted linear accelerator (linacs) (Saturne, General Electric). Median minimal dose delivered to the gross disease was 16.2 Gy (11.8-23), and median maximal dose 21.2 Gy (14- 42). RESULTS: Median follow-up was 9 months (1-57). Five metastases recurred. One- and 2-year metastasis local control rates were 91.3 and 84% and 1- and 2-year brain control rates were 65 and 57%, respectively. Six patients died of brain metastasis evolution, and three of leptomeningeal carcinomatosis. One- and 2-year overall survival rates were 31 and 28%, respectively. According to univariate analysis, KPS, RPA class, SIR score and interval between WBRT and radiosurgery were prognostic factors of overall survival and brain free-disease survival. According to multivariate analysis, RPA was an independent factor of overall survival and brain free-disease survival, and the interval between WBRT and radiosurgery longer than 14 months was associated with longer brain free disease survival. Side effects were minimal, with only two cases of headaches and two of grade 2 alopecia. CONCLUSION: Salvage radiosurgery of metastasis recurring after whole brain irradiation is an effective and accurate treatment which could be proposed to patients with a KPS>70 and a primary tumour controlled or indolent. We recommend that a dose not exceeding 14 Gy should be delivered to an isodose representing 70% of the maximal dose since local control observed rate was similar to that previously published in literature with upper dose and side effects were minimal. PMID- 11410306 TI - Differential effects of buthionine sulphoximine in hypoxic and non-hypoxic regions of human cervical carcinoma xenografts. AB - BACKGROUND AND PURPOSE: Recently we reported increased glutathione (GSH) levels in hypoxic regions of ME 180 and SiHa cervical cancer xenografts. Since this association might act synergistically to protect from radiotherapy, we examined the differential effects of the GSH depleting agent buthionine suiphoximine (BSO) in relation to tumor oxygenation. MATERIALS AND METHODS: The nitroimidazole EF5 was used to label tumor hypoxia. GSH levels were determined in cryostat sections using a sensitive HPLC assay and in parallel sections using fluorescence image analysis. Using a dual-labeling method, GSH levels were determined selectively in hypoxic and non-hypoxic tumor regions. RESULTS: GSH levels were higher in hypoxic than in non-hypoxic regions of cervical carcinoma xenografts. Treatment with BSO produced a more pronounced GSH depletion in regions of hypoxia, resulting in similar post-treatment levels in hypoxic and non-hypoxic areas. CONCLUSIONS: BSO effectively depletes GSH in hypoxic microregions of tumors. These findings suggest a potential role for BSO as an adjunct to radiotherapy in cervical cancer patients. PMID- 11410307 TI - Normal cellular radiosensitivity in an adult Fanconi anaemia patient with marked clinical radiosensitivity. AB - BACKGROUND: Fanconi anaemia is a rare disease associated with cellular sensitivity to chemicals (e.g. mitomycin C and diepoxybutane); variable but mild cellular radiosensitivity has also been reported. MATERIALS AND METHODS: A 32 year-old patient with Fanconi anaemia and tonsillar carcinoma, treated by radiotherapy, was found to exhibit profound clinical radiosensitivity. Confluent, ulcerating oropharyngeal mucositis developed after a conventionally fractionated dose of 34Gy and healing was incomplete by 2 months after cessation of therapy. RESULTS: Cellular radiosensitivity assays and RPLD studies from this patient did not suggest any major detectable radiosensitivity. CONCLUSION: There is a discrepancy between the observed clinical radiosensitivity and the usual "predictive" radiosensitivity assays in this patient with Fanconi anaemia. PMID- 11410308 TI - Quantitative status of resources for radiation therapy in Asia and Pacific region. AB - PURPOSE: Resources for radiation therapy in Asian and Pacific countries were analyzed to obtain a better understanding of the status of radiation oncological practice in the region. METHODS AND MATERIALS: The data were obtained mainly through surveys on the availability of major equipment and personnel which were conducted through an International Atomic Energy Agency regional project. The study included 17 countries in South Asia, South East Asia, East Asia and Australasia. Data were related to national populations and economic and a general health care indices. RESULTS: Large differences in equipment and personnel among countries were demonstrated. The availability of both teletherapy and brachytherapy was related to the economic status of the countries. The shortage of teletherapy machines was evident in more countries than that of brachytherapy. Many departments were found to treat patients without simulators or treatment planning systems. The number of radiation oncologists standardized by cancer incidence of a country did not correlate well with economic status. CONCLUSIONS: There were significant deficiencies in the availability of all components of radiation therapy in the analyzed countries. The deficiencies were linked predominantly to the economic status of the country. Cognisance should be taken of the specific shortfalls in each country to ensure that expansion or any assistance offered appropriately match its needs and can be fully utilized. The information on the resources currently available for radiation oncological practice in the region presented in this paper provides a valuable basis for planning of development aid programs on radiation therapy. PMID- 11410309 TI - Clinical research in developing countries--ethical dilemmas. PMID- 11410310 TI - The effect of breathing and set-up errors on the cumulative dose to a lung tumor. AB - BACKGROUND AND PURPOSE: To assess the impact of both set-up errors and respiration-induced tumor motion on the cumulative dose delivered to a clinical target volume (CTV) in lung, for an irradiation based on current clinically applied field sizes. MATERIALS AND METHODS: A cork phantom, having a 50 mm spherically shaped polystyrene insertion to simulate a gross tumor volume (GTV) located centrally in a lung was irradiated with two parallel opposed beams. The planned 95% isodose surface was conformed to the planning target volume (PTV) using a multi leaf collimator. The resulting margin between the CTV and the field edge was 16 mm in beam's eye view. A dose of 70 Gy was prescribed. Dose area histograms (DAHs) of the central plane of the CTV (GTV+5 mm) were determined using radiographic film for different combinations of set-up errors and respiration-induced tumor motion. The DAHs were evaluated using the population averaged tumor control probability (TCP(pop)) and the equivalent uniform dose (EUD) model. RESULTS: Compared with dose volume histograms of the entire CTV, DAHs overestimate the impact of tumor motion on tumor control. Due to the choice of field sizes a large part of the PTV will receive a too low dose resulting in an EUD of the central plane of the CTV of 68.9 Gy for the static case. The EUD drops to 68.2, 66.1 and 51.1 Gy for systematic set-up errors of 5, 10 and 15 mm, respectively. For random set-up errors of 5, 10 and 15 mm (1 SD), the EUD decreases to 68.7, 67.4 and 64.9 Gy, respectively. For similar amplitudes of respiration-induced motion, the EUD decreases to 68.8, 68.5 and 67.7 Gy, respectively. For a clinically relevant scenario of 7.5 mm systematic set-up error, 3 mm random set-up error and 5 mm amplitude of breathing motion, the EUD is 66.7 Gy. This corresponds with a tumor control probability TCP(pop) of 41.7%, compared with 50.0% for homogeneous irradiation of the CTV to 70 Gy. CONCLUSION: Systematic set-up errors have a dominant effect on the cumulative dose to the CTV. The effect of breathing motion and random set-up errors is smaller. Therefore the gain of controlling breathing motion during irradiation is expected to be small and efforts should rather focus on minimizing systematic errors. For the current clinically applied field sizes and a clinically relevant combination of set-up errors and breathing motion, the EUD of the central plane of the CTV is reduced by 3.3 Gy, at maximum, relative to homogeneous irradiation of the CTV to 70 Gy, for our worst case scenario. PMID- 11410312 TI - Purification and characterization of White Spot syndrome virus (WSSV) produced in an alternate host: crayfish, Cambarus clarkii. AB - Penaeid shrimp is the natural host of White Spot Syndrome Virus (WSSV) that can cause high mortality in the infected hosts. Attempts to obtain sufficient amounts of purified intact WSSV for characterization have been unsuccessful. Using crayfish, Cambarus clarkii as a proliferation system, a large amount of infectious WSSV was reproduced and intact WSSV viral particles were purified with a new isolation medium by ultra-centrifugation. Purified WSSV particles were very sensitive to organic solvents and the detergent, Triton X-100. The size of the rod-shape, somewhat elliptical, intact WSSV was 110-130x260-350 nm with a long, tail-like envelope extension. The naked viral nucleocapsid was about 80x350 nm, and it possessed 15 spiral and cylindrical helices composed of 14 globular capsomers along its long axis, and a 'ring' structure at one terminus. Distinct WSSV genome DNA patterns were obtained when the purified genomic dsDNA of WSSV was digested with five different restriction enzymes (HindIII, XhoI, BamHI, SalI, and SacI). In addition, at least 13 major and distinct protein bands could be observed when purified intact WSSV viruses were separated by SDS-PAGE followed by Coomassie Brilliant R-250 staining. The estimated molecular weights of these proteins were 190, 84, 75, 69, 68, 58, 52, 44, 28, 27.5, 23, 19, and 16 kD, respectively. Both the 44 and 190 kD proteins were easily removed if the hemolymph from the WSSV infected crayfish was transiently treated with 1% Triton X-100 before it was subjected to gradient centrifugation, indicating that both of them are located on the surface of the viral envelope. These characteristics are consistent with WSSV isolated from the penaeid shrimp. PMID- 11410313 TI - Changes in BiP (GRP78) levels upon HSV-1 infection are strain dependent. AB - BiP (grp78) is a chaperone protein which can also regulate the unfolded protein response of the cell. Levels of BiP increased in cells infected by the small plaque producing, cell associated, neuroinvasive strains of HSV-1 (SP7, 490) but decreased in cells infected with KOS, a large plaque, attenuated strain. BiP protein synthesis continued early in infection and BiP was sequestered and its degradation was limited during SP7 infection. BiP protein synthesis stopped and the protein was degraded in KOS infected cells. These viral strain dependent differences in BiP concentration may influence other aspects of the viral interaction with the target cell and its host. PMID- 11410314 TI - Using HIV-1 sequence variability to explore virus biology. AB - Human immunodeficiency virus type 1 (HIV-1) only recently established an epidemic world-wide infection in the human population. The virus persists in the human host through active replication and is able to avoid clearance by the immune system. Active replication is an important component of the rapid evolutionary potential of HIV-1, a potential which manifests itself in the evolution of immune escape variants, drug resistant variants, and variants with the ability to use different cell surface coreceptors in conjunction with CD4. Multiple zoonotic introductions, compartmentalization of virus replication in the body, and genetic bottlenecks associated with sampling during transmission, antiretroviral therapy, and geographic and/or host population isolation further contribute to the range of sequences present in extant viruses. The sum of the history of all of these phenomena is reflected in HIV-1 sequence variability, and most of these phenomena are ongoing today. Here we review the use of HIV-1 sequence variability to explore its underlying biology. PMID- 11410315 TI - Tick-borne encephalitis virus NS1 glycoprotein during acute and persistent infection of cells. AB - Tick-borne encephalitis virus (TBEV) was propagated in porcine embryo kidney (PS) cells until 48 h whereas human kidney (RH) cells maintained the virus persistence during at least 2 months. One of possible reasons of flavivirus chronic infection might be abnormal NS1 gene expression. Immunoblotting with monoclonal antibodies (MAbs) revealed the similarity of the intracellular and secreted NS1 nonstructural glycoprotein size and linear antigenic determinants in both the infected cell lines. However, according to the competitive binding of MAbs with the TBEV NS1 extracellular glycoprotein, its contiguous epitopes differed for acute or persistent infection. To map the TBEV NS1 glycoprotein antigenic determinants its recombinant analogues were used. All the studied MAbs could bind with the full-length NS1 recombinant protein. Deletion of the TBEV NS1 gene internal region resulted in defective NS1d1 protein without the region between 269 and 333 a.a. Lack of NS1d1 binding with 20B4 MAb and diminished binding with 22H8 and 17C3 MAbs permitted to map their antigenic determinants within or nearby deleted region, respectively. Interaction of other MAbs with the NS1 and NS1d1 recombinant proteins did not differ, suggesting that their epitopes were located in the region of N-terminal 268 a.a. or C-terminal 19 a.a. of the TBEV NS1 protein. The second NS1d2 truncated protein contained the first N-terminal 33 a.a. of the TBEV NS1 protein and was able to bind with 29G9 MAb. Taken together the data stand for the differences in the N-terminal structure of the TBEV NS1 multimers secreted from the acute and persistent infected cells whereas the intracellular and secreted monomer processing was the same. The modified NS1 protein oligomers in the RH cellular line might slow virus replication and could result in the TBEV persistence. PMID- 11410316 TI - Genetic analyses of feline foamy virus isolates from domestic and wild feline species in geographically distinct areas. AB - To know the genetic diversities and phylogenetic relationship among feline foamy virus (FeFV) isolates from domestic cats (Felis catus) and FeFV-related viruses from the Iriomote cats (Felis iriomotensis) and leopard cats (Felis bengalensis) in geographically distinct areas, we sequenced a partial gag-pol region of 17 strains and a partial env region of nine strains, and the U3 region of long terminal repeat of three strains of the viruses. FeFV-related viruses from the feral cats were quite similar to the FeFV from domestic cats in the sequenced regions. In the partial gag region, the identities of nucleotide sequences among the isolates were from 94 to 99%. In the partial env gene, the isolates were divided into two distinct genotypes (F17- and FUV-types) as reported by Winkler et al. (Virology 247 (1999) 144-151). More than 94% nucleotide identities were observed in the env region within a particular env genotype and about 75% nucleotide identities were noted between the two genotypes. PMID- 11410317 TI - The H1 double-stranded RNA genome of Ustilago maydis virus-H1 encodes a polyprotein that contains structural motifs for capsid polypeptide, papain-like protease, and RNA-dependent RNA polymerase. AB - The Ustilago maydis viral (UmV) genome consists of three distinct size groups of double-stranded RNA (dsRNA) segments: H (heavy), M (medium), and L (light). The H segments have been suggested to encode all essential viral proteins, but without any molecular evidences. As a preliminary step to understand viral genomic organization and the molecular mechanism governing gene expression in UmV, we determined the complete nucleotide sequence of the H1 dsRNA genome in P1 viral killer subtype. The H1 dsRNA genome (designated UmV-H1) contained a single open reading frame that encodes a polyprotein of 1820 residues, which is predicted to be autocatalytically processed by a viral papain-like protease to generate viral proteins. The amino-terminal region is the capsid polypeptide with a predicted molecular mass of 79.9 kDa. The carboxy-terminal region is the RNA-dependent RNA polymerase (RDRP) that has a high sequence homology to those of the totiviruses. The H2 dsRNA also encodes a distinct RDRP, suggesting that UmV is a complex virus system like the Saccharomyces cerevisiae viruses ScV-L1 and -La. PMID- 11410318 TI - Short-term exposure to estrogen and progesterone induces partial protection against N-nitroso-N-methylurea-induced mammary tumorigenesis in Wistar--Furth rats. AB - The lifetime protective effect of a full term pregnancy for breast cancer is a reproducible and consistent finding in human beings and in rodent models. The duration of pregnancy necessary to confer protection has yielded contradictory results. As the administration of estrogen and progesterone mimics the full-term pregnancy effect on conferring protection, we examined whether short-term exposure to estrogen and progesterone confers protection against N-nitroso-N methylurea-induced mammary carcinogenesis in Wistar--Furth rats. The results reported herein show that treatment of rats with estrogen or progesterone alone for 21 days does not confer protection, but a 10-day exposure to the same concentrations of estrogen and progesterone induced a partial protective effect. The significance of these results are discussed in terms of the contradictory results in the literature and the role of morphological differentiation in conferring the protective effect. PMID- 11410319 TI - Significance of contracted cholecystitis lesions as high risk for gallbladder carcinogenesis. AB - A precancerous change has been identified incidentally in resected specimens from patients who have undergone cholecystectomy. We focused on chronic cholecystitis, showing a thick and sclerotic wall caused by recurrent inflammation, e.g. contracted cholecystitis, and examined the malignant potential of these lesions. We studied 88 patients who had undergone cholecystectomy. Contracted cholecystitis was diagnosed, using our criteria, in 28 of these cases. Ordinary chronic cholecystitis was diagnosed in 50 cases and gallbladder carcinoma in ten cases. We examined the expression of p53, Ki-67, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) immunohistochemically. Severe dysplasia or carcinoma in situ in a very small portion of the specimen was identified with hematoxylin-eosin staining in four cases (14.3%) of contracted cholecystitis. These specimens revealed a positive expression of not only p53, but also Ki-67, iNOS, and COX-2. Statistical significance was shown among the three disease groups in terms of the incidence of p53 overexpression, respectively (P<0.05). The results of this study suggest that contracted cholecystitis could be an early change leading to carcinogenesis. PMID- 11410320 TI - Cancer chemopreventive agents, 4-phenylcoumarins from Calophyllum inophyllum. AB - In a search for anti-tumor-promoting agents, we carried out a primary screening of ten 4-phenylcoumarins isolated from Calophyllum inophyllum L. (Guttiferae), by examining their possible inhibitory effects on Epstein--Barr virus early antigen (EBV-EA) activation induced by 12-O-tetradecanoylphorbol-13-acetate in Raji cells. All of the compounds tested in this study showed inhibitory activity against EBV, without showing any cytotoxicity. Calocoumarin-A (5) showed more potent activity than any of the other compounds tested. Furthermore, calocoumarin A (5) exhibited a marked inhibitory effect on mouse skin tumor promotion in an in vivo two-stage carcinogenesis test. The results of the present investigation indicate that some of these 4-phenylcoumarins might be valuable as potential cancer chemopreventive agents (anti-tumor-promoters). PMID- 11410321 TI - Glutathione S-transferase M1 and T1 null genotypes and the risk of gastric and colorectal cancers. AB - Several polymorphic glutathione S-transferase (GST) enzymes are involved in the detoxification of active metabolites of many potential carcinogens and may therefore be important in modulating susceptibility to cancers. GSTM1 and GSTT1 are polymorphic, and the null alleles result in a lack of corresponding enzyme activities. Previous studies demonstrated that the GSTM1 and GSTT1 null genotypes correlated with an increased risk of developing some cancers. In this study, we determined GSTM1 and GSTT1 polymorphisms in a population of 131 healthy controls from the south of Iran, 46 patients with colorectal cancers, and 42 patients with gastric cancer. The gastric cancer risk statistically increased due to the GSTM1 null genotype (odds ratio (OR)=2.3, 95% confidence interval (CI): 1.15--4.95). On the other hand, the GSTT1 null genotype in gastric cancer and null genotypes of GSTM1 and GSTT1 in colorectal cancer were not statistically significant. Moreover, individuals showing the GSTM1 and GSTT1 null genotypes might exhibit a greater predisposition to gastric (OR=3.31, 95% CI: 1.14--9.57) and colorectal (OR=2.73, 95% CI: 0.94--7.95, P=0.07) cancers. PMID- 11410322 TI - EGFR tyrosine kinase inhibitor AG1478 inhibits cell proliferation and arrests cell cycle in nasopharyngeal carcinoma cells. AB - Nasopharyngeal carcinoma (NPC), which occurs with a high incidence in southern China and southeast Asia, is of epithelial origin with overexpression of EGF receptor. To study the effect of inhibition of EGFR signaling on nasopharyngeal carcinoma cell proliferation and cell cycle distribution, EGFR tyrosine kinase inhibitor AG1478 was employed to treat Nasopharyngeal Carcinoma CNE2 cells. The results showed that AG1478 inhibited proliferation of CNE2 cells. Immunoblot showed that AG1478 inhibited EGFR phosphorylation in CNE2 cells without reduced expression of EGFR protein. The activation of Akt and MAPK which are downstream molecules of EGFR signaling pathway, were also inhibited by AG1478. AG1478 induced cell cycle arrest in G1 phase, and the levels of protein p27 were significantly up-regulated. We concluded that inhibition of the EGFR signaling induced cell cycle arrest in G1 phase in CNE2 cells and p27 up-regulation was involved in this process. The EGFR kinase specific inhibitor is of potential to be developed into drugs for NPC treatment. PMID- 11410323 TI - Evaluation of protoporphyrin IX production, phototoxicity and cell death pathway induced by hexylester of 5-aminolevulinic acid in Reh and HPB-ALL cells. AB - Production of protoporphyrin IX (PpIX) in human B-cell leukemia cell line (Reh) and T-cell lymphoma cell line (HPB-ALL) was studied by flow cytometry after incubation with 5-aminolevulinic acid (ALA) or its hexylester in vitro. Cell survival and cell death pathway were also investigated in these two cell lines by cell growth curves, flow cytometry, and electron microscopy after ALA hexylester mediated photodynamic therapy. Both ALA and its hexylester could induce PpIX production in the two cell lines, but ALA hexylester was about 100 times more efficient than ALA. Reh cells appear to be more sensitive than HPB-ALL cells to ALA hexylester-mediated phototoxicity. Apoptosis was the major cell death pathway of Reh cells, while necrosis played a major role in the case of HPB-ALL cells. PMID- 11410324 TI - Polo-like kinase (PLK) expression in endometrial carcinoma. AB - Polo-like kinase (PLK) is a cell cycle-regulated, cyclin-independent serine/threonine protein kinase. Recent reports have shown a critical role for PLK during tumorigenesis. To explore whether PLK plays a general role as a tumor marker of endometrial carcinomas, we examined the expression of PLK mRNA and protein in endometrial carcinomas and normal endometrium, and analyzed the relationship between PLK protein expression and malignant potential. We found that PLK mRNA was expressed in all specimens from endometrial carcinoma patients using RT-PCR methods, although some specimens from normal endometria were negative. Immunohistochemically, most of the PLK was found in the cytoplasm (around the nucleus), and partly in the nucleus of endometrial carcinoma glands and also secreted tissues from endometrial carcinoma glands. PLK was expressed at the basement membrane of carcinoma glands and partly expressed in the head portion of papillary carcinoma tissues. There was a significant correlation between percentages of PLK-positive cells and histological grade of endometrial carcinoma (P<0.0001). However, the expression of proliferating cell nuclear antigen and Ki-67 was independent of PLK expression. Moreover, we noted that PLK is strongly expressed in invading carcinoma cells. PLK expression could reflect the degree of malignancy and proliferation in endometrial carcinoma. Thus, in addition to being of diagnostic value, modulation of PLK activity in the tumors by chemotherapeutic agents or gene therapy may prove to be of therapeutic value. PMID- 11410325 TI - Role of cAMP-dependent protein kinase in the regulation of DNA repair. AB - Enhanced DNA repair is an important factor in drug resistance in cancer. Using cell-free extracts derived from the fission yeast, Schizosaccharomyces pombe, we demonstrate in an in vitro system DNA repair system that increased cAMP levels, which activates cAMP-dependent protein kinase (PKA), inhibits repair of ultraviolet (UV)-damaged DNA. Supplementing the cell-free system with the catalytic kinase subunit of PKA also inhibits DNA repair. In contrast, addition of the PKA inhibitor H-89 enhances repair activity. These results show that PKA regulates DNA repair synthesis, thus implicating the cAMP signaling pathway in DNA damage response and repair of UV-damaged DNA lesions. PMID- 11410326 TI - Genetic alterations in gallbladder adenoma, dysplasia and carcinoma. AB - Adenoma and dysplasia in the gallbladder (GB) have been reported as precancerous lesions, but the genetic evidence of this is not clearly defined. The purpose of this study was to analyze the frequencies of K-ras, p53, and p16 gene mutations, of microsatellite instability (MI) and of loss of heterozygosity (LOH) in GB cancer, dysplasia, and adenoma. Tissues from 15 GB cancers, five dysplasias around cancerous tumors, and three adenomas were collected prospectively. The mutation rates of K-ras, p53, and p16 were 20.0, 35.7, and 30.7%, respectively, in GB cancers. However, no mutations were found in dysplasia or adenoma. Reduced staining for p16 was seen in 23% of carcinomas. All of the GB carcinomas and four out of five (80%) of the dysplasias showed LOH in a minimum of one locus, but one out of three (33%) cases of adenoma displayed LOH in only one locus. All of the loci of LOH in the dysplasias, except one, showed the same patterns of allelic loss as the adjacent carcinomas. Only one dysplasia showed multiple MI. In conclusion, multiple LOH may be associated with the development of dysplasia and the malignant transformation of GB carcinoma. Gene alterations of K-ras, p53, and p16 are important steps in the malignant changes of dysplasia. However, MI seems to have only a limited role in GB cancer development. PMID- 11410327 TI - p53 gene alteration and protein expression in Iranian women with infiltrative ductal breast carcinoma. AB - Genetic alterations in the p53 tumor suppressor gene with or without protein overexpression have been reported to be associated with sporadic breast cancer. To assess the role of p53 in infiltrative ductal breast carcinoma among Iranian patients, p53 protein expression and p53 gene mutations were studied by immunohistochemical analysis and single-strand conformation polymorphism, respectively. The p53 protein was expressed in 25 out of 51 (49%) tumors and p53 gene mutations were detected in 17 out of 37 (46%) tested tumors. No significant correlation was observed between p53 gene mutations and p53 protein expression. There was no significant correlation between p53 abnormalities (mutation and expression) and tumor size, histological grade, nodal status, and progesterone receptor expression. However, a non-statistically significant trend of association (P=0.07) was observed between p53 gene mutations and lack of estrogen receptor. The high percentage of alterations both in p53 gene and protein among southern Iranian breast cancer patients suggests that p53 is probably one of the genes involved in sporadic breast cancer in this area. PMID- 11410328 TI - Animal model of para-aortic lymph node metastasis. AB - The purpose of this study was to establish a model of experimental lymph node metastasis by intra-rectal implantation of human cancer cells in nude mice. Four types of human cancer cell lines (TE-1, MKN-45, HT-29, and MIAPaca-2) were investigated. Tumor cells suspended in Matrigel were injected into the submucosal layer of the rectum. All cancer cell lines produced locally aggressive rectal tumors and, subsequently, para-aortic lymph node metastasis. We were unable to produce other distant metastases in the dying state in such locations as the liver, spleen, lung, and peritoneum. However, using this method, we were able to evaluate the effect of the anti-cancer agent uracil/tegafur (UFT) on primary tumor growth and lymph node metastasis. Oral intake of UFT significantly suppressed implanted tumor volume and inhibited lymph node metastasis. We expect that the process of lymph node metastasis shown in this model will be studied as an experimental model of lymph node metastasis simulating human cancers. PMID- 11410329 TI - Interleukins-6 and -11 expression in primary breast cancer and subsequent development of bone metastases. AB - Breast cancers frequently metastasize to bone where they often cause extensive tumor-induced osteoclast-mediated osteolysis. Interleukin-6 (IL-6) and IL-11 are two cytokines exhibiting osteolytic properties through their potent stimulation of osteoclast formation. We investigated the expression of IL-6 and IL-11 in 99 invasive primary breast tumors by immunohistochemistry and in situ hybridization, respectively. We examined their potential as predictive factors for further development of bone metastases. 52/90 (57%) of tumor samples showed IL-6 cytoplasmic immunostaining. There was no significant association between IL-6 status and any of the classical prognostic factors. 15/89 (17%) of the tumor samples expressed IL-11 mRNA. A positive IL-11 mRNA status was associated with a low tumor grade (P=0.05). Tumors expressing IL-11 mRNA had a statistically significant (P=0.002) higher rate of bone metastases occurrence (12/15, 80%) than IL-11 negative tumors (27/74, 37%). Such association was not found for IL-6. Our findings demonstrate for the first time IL-11 gene expression in some primary invasive breast tumors and suggest the potential of this cytokine as possible biological predictive factor for the development of bone metastases. PMID- 11410330 TI - Recognition and clearance of methoxypoly(ethyleneglycol)2000-grafted liposomes by macrophages with enhanced phagocytic capacity. Implications in experimental and clinical oncology. AB - Intravenous injection of an endotoxin-free solution of poloxamine-908 to rats can enhance the phagocytic clearance capacity of tissue macrophages, particularly those of the liver and the spleen. Such stimulated cells were able to clear a significant portion of intravenously injected methoxypoly(ethyleneglycol)2000 liposomes (mean size of 87 nm), labelled with technetium-99m via the N hydroxysuccinimidyl hydrazine nicotinate hydrochloride derivative of distearoyl phosphatidylethanolamine, within 4 h post administration. These liposomes, otherwise, exhibit long circulatory behaviour in control animals, with poor localization to the liver and spleen. We suggest that such technetium-99m labelled engineered vesicles may be of aid for detection of the liver and spleen macrophages with enhanced phagocytic clearance capacity by gamma scintigraphy. Alterations in the phagocytic activity of liver and spleen macrophages is known to occur during cancer. Therefore, such diagnostic procedures may prove useful for patient selection or for monitoring the progress of treatment with long circulating nanoparticles carrying anti-cancer agents, thus minimizing damage to this important line of body's defence cells, and are discussed. PMID- 11410331 TI - In vivo image-guided (1)H-magnetic resonance spectroscopy of the serial development of hepatocarcinogenesis in an experimental animal model. AB - Histology on a core or open biopsy is considered the gold standard for the diagnosis of tumours. While the non-invasive technique of magnetic resonance imaging can direct some of the decision diagnostic making, it has limitations and disadvantages, that can be partly overcome with the use of in vivo magnetic resonance spectroscopy (MRS). In vivo MRS is able to provide a specific biochemical profile on tumour tissue, compared with normal tissue. The capability of this technique is demonstrated here by the long-term development of hepatocellular carcinoma in an animal model. It allows the observation of the biochemical changes that occur in tumour tissue during its progression from preneoplastic nodules to hepatocellular carcinoma. Specifically the changes in the lipid profiles of tumour tissue at various stages of development are observed with proton ((1)H) MRS. Significant increases occurred in the lipid acyl chain methylene and methyl hydrogens during the early developmental stages of hepatocarcinogenesis, whereas during later stages associated with tumour development there was a significant increase in the levels of olefinic acyl chain hydrogens from unsaturated lipids. It is anticipated that this model will precede the application of the same technology to the non-invasive diagnosis and grading of human hepatocellular carcinoma. PMID- 11410332 TI - Thioredoxin reductase and cytoplasmic glutathione peroxidase activity in human foetal and neonatal liver. AB - Cytosolic thioredoxin reductase (TR) is an FAD-containing homodimeric selenoenzyme which, together with thioredoxin (Trx) and NADPH, forms a powerful oxidoreductase system. Cytoplasmic glutathione peroxidase (GPX-1) is a selenoprotein with antioxidant activity. The TR/Trx system has been associated with cellular processes including regulation of cell growth, and modification of activity of transcription factors. TR may also act as an antioxidant. We have measured TR activity, TR concentration, and GPX-1 activity in human hepatic cytosols from foetuses and neonates. The concentration of TR was significantly greater (P<0.05) in foetal (43.6, 37.9-50.8 microg/g protein, median, interquartile range) than in neonatal liver (11.6, 8.70-15.0 microg/g). This was also true of TR activity which was 2.1, 1.8-2.5 U/g protein in foetal, and 0.65, 0.44-0.74 U/g protein in neonatal liver (P<0.0005). Similarly, GPX-1 activity was significantly higher (P<0.005) in the foetal (199.7, 144.0-227.9 U/g protein) than in neonatal (77.0, 58.4-110.3 U/g protein) hepatic cytosol. Overall, foetal liver expressed approx. 3-fold higher activities of TR and GPX-1 than neonatal liver. PMID- 11410333 TI - Chemo-enzymatic synthesis of a bioactive peptide containing a glutamine-linked oligosaccharide and its characterization. AB - A bioactive peptide containing a glutamine-linked oligosaccharide was chemo enzymatically synthesized by use of the solid-phase method of peptide synthesis and the transglycosylation activity of endo-beta-N-acetylglucosaminidase. Substance P, a neuropeptide, is an undecapeptide containing two L-glutamine residues. A substance P derivative with an N-acetyl-D-glucosamine residue attached to the fifth or sixth L-glutamine residue from the N-terminal region was chemically synthesized. A sialo complex-type oligosaccharide derived from a glycopeptide of hen egg yolk was added to the N-acetyl-D-glucosamine moiety of the substance P derivative using the transglycosylation activity of endo-beta-N acetylglucosaminidase from Mucor hiemalis, and a substance P derivative with a sialo complex-type oligosaccharide attached to the L-glutamine residue was synthesized. This glycosylated substance P was biologically active, although the activity was rather low, and stable against peptidase digestion. The oligosaccharide moiety attached to the L-glutamine residue of the peptide was not liberated by peptide-N(4)-(N-acetyl-beta-D-glucosaminyl) asparagine amidase F. PMID- 11410334 TI - Saponin permeabilization of rough microsomes from rat liver reveals a novel prothrombin pool. AB - Saponin permeabilization of rough microsomes in the presence of high salt revealed a novel pool of prothrombin associated by ionic interactions to the microsomal membrane. The lumenal content was obtained by treating rough microsomes with 0.32% saponin in a low salt (0.05 M KCl) buffer. By a subsequent treatment with 0.32% saponin in a slightly alkaline high salt buffer a fraction of peripherally associated membrane prothrombin was released from rough microsomes. Finally, the membrane-bound fraction was solubilized with 2.5% Triton X-100. The lumenal content fraction, the peripherally membrane-associated and the membrane-bound fraction from normal rats contained 55%, 29% and 16% of the total rough microsomal prothrombin, respectively. The corresponding fractions from warfarin-treated rats contained 86%, 5% and 9% of the total prothrombin. Following (14)C-gamma-carboxylation of intact microsomes for 30 min, the novel membrane-associated and the membrane-bound pool contained 42% and 33%, respectively, of labeled prothrombin. A similar distribution was found with warfarin-treated rats. PMID- 11410335 TI - Expression, purification, characterization and homology modeling of active Akt/PKB, a key enzyme involved in cell survival signaling. AB - Akt is a serine/threonine kinase that plays a critical role in cell survival signaling and its activation has been linked to tumorigenesis. Up-regulation of Akt as well as its upstream regulator phosphatidylinositol-3 kinase (PI3K) has been found in many tumors and the negative regulator of this pathway PTEN/MMAC is a tumor suppressor. As a target for drug discovery, we have expressed and purified an active Akt1 enzyme from a recombinant baculovirus-infected Sf9 cell culture. Coexpression of Akt1 with the catalytic subunit of PI3K or treatment with okadaic acid during expression was found to generate an active enzyme in the insect cell culture system. We have optimized the kinase activity and developed a simple quantitative kinase assay using biotinylated peptide substrates. Using the purified active enzyme, we have characterized its physical, catalytic and kinetic properties. Since Akt is closely related to protein kinase C (PKC) and protein kinase A, the issue of obtaining selective inhibitors of this enzyme was addressed by comparison of the structures of catalytic domains of Akt and PKC, derived by homology modeling methods. A number of amino acid differences in the ATP binding regions of these kinases were identified, suggesting that selective inhibitors of Akt can be discovered. However, the ATP binding regions are highly conserved in the three isoforms of Akt implying that the discovery of isoform selective inhibitors would be very challenging. PMID- 11410336 TI - Frequency-dependent effects of ELF magnetic field on chromatin conformation in Escherichia coli cells and human lymphocytes. AB - The effects of magnetic fields of extremely low frequency (ELF, 21 microT r.m.s.) on cells of different Escherichia coli K12 strains and human lymphocytes were studied by the method of anomalous viscosity time dependence (AVTD). Within the frequency range of 6-24 Hz, two resonance-type frequency windows with maximal effects at 9 Hz and 16 Hz were observed in response of GE499 strain. Only one frequency window with maximum effect at 8.5 Hz was found for GE500 cells. These data along with previously obtained for two other E. coli strains, AB1157 and EMG2, indicate that frequency windows are dependent on genotype of cells exposed to ELF. Resonance-type effects of ELF were also observed in human lymphocytes in frequency windows around 8 and 58 Hz. These ELF effects differed significantly between studied donors, but were well reproducible in independent experiments with lymphocytes from the same donors. The frequency windows in response of E. coli strains and human lymphocytes to ELF significantly overlapped suggesting that the same targets may be involved in this response. We compared the frequency windows with predictions based on the ion cyclotron resonance (ICR) model and the magnetic parametric resonance model. These models predicted effects of ELF magnetic fields at the 'cyclotron' frequencies of some ions of biological relevance. According to the ICR model, ELF effects should be also observed at harmonics of cyclotron frequencies and, contrary, parametric resonance model predicted effects at subharmonics. While we observed coincidence of each experimental resonance frequency with predictions of one of these two models, all experimentally defined effective frequency windows were in good agreement with relatively narrow frequency ranges of both harmonics and subharmonics for natural isotopes of Na, K, Ca, Mg, and Zn ions. The experimental data support idea that both harmonics and subharmonics of several biologically important ions are involved in frequency-dependent ELF effects in cells of different types. PMID- 11410337 TI - Oxidative breakdown and conversion of urocanic acid isomers by hydroxyl radical generating systems. AB - cis-Urocanic acid (cis-UCA), formed from trans-urocanic acid (trans-UCA) by photoisomerization, has been shown to mimic suppressive effects of UV on the immune system. It is our hypothesis that UCA oxidation products in the skin play a role in the process of immunosuppression. Recently, both UCA isomers were found to be good hydroxyl radical scavengers and in this context we investigated the formation of products resulting from the interaction of hydroxyl radicals with UCA. Hydroxyl radicals were generated by (1) UV/H(2)O(2) (photooxidation), (2) ferrous ions/H(2)O(2) (Fenton oxidation) and (3) cupric ions/ascorbic acid. Oxidation products were identified by spectrometric methods and assessed by reversed-phase HPLC analysis. The photooxidation of UCA was induced by UV-B and UV-C, but not by UV-A radiation. Photooxidation and Fenton oxidation of trans UCA, as well as of cis-UCA yielded comparable chromatographic patterns of UCA oxidation products. Several of the formed products were identified. The formation of three identified imidazoles was shown in UV-B exposed corneal layer samples, derived from human skin. PMID- 11410338 TI - Some features of intestinal absorption of intact fibrinolytic enzyme III-1 from Lumbricus rubellus. AB - In order to investigate whether earthworm fibrinolytic enzyme III-1 (EFE-III-1) isolated from Lumbricus rubellus is capable of transporting into blood through intestinal epithelium and keeping its biological function in circulation, we have raised an antibody against EFE-III-1. The immunological results showed that 10 15% of intact EFE-III-1 was absorbed by the intestinal epithelium with the incubation chamber method [Vilhardt and Lundin, Acta Physiol. Scand. 126 (1986) 601-607]. The enzyme could be detected in the intestinal epithelial cells by immunohistochemistry. Furthermore, immunoreactive intact EFE-III-1 was found in serum or plasma after intraperitoneal injection of rats. Approx. 10% of the full size enzyme could transport through the intestinal epithelium. The maximum remaining activity in blood could be assayed around 60 min after the intraperitoneal injection. PMID- 11410339 TI - Reduced glutathione protect cells from ouabain toxicity. AB - It is widely accepted that a prolonged ouabain blockade of the Na(+),K(+)-ATPase makes cells detach from each other and from the substrate, leading to their death and that cellular resistance to ouabain is due to the presence of isoforms of Na(+),K(+)-ATPase with low affinity to this glycoside. In the present work the effect of reduced glutathione in the response of two types of renal cells to ouabain: MDCK, a ouabain-sensitive cell line and Ma104, a ouabain-resistant one, was studied. Glutathione protected MDCK cells from ouabain toxicity and inhibition of glutathione synthesis by L-buthionine-S,R-sulfoximine sensitized Ma104 cells to ouabain. As glutathione is involved with multidrug resistance (MDR) in cells expressing the multidrug resistance-related protein MRP1 and as Ma104 cells have a MDR phenotype, it was investigated whether Ma104 cells express this protein. The expression of the MRP1-mRNA in Ma104 cells was detected by reverse transcriptase-polymerase chain reaction and ribonuclease protection assay, and the protein was detected by Western blotting and immunofluorescence. Treatment of Ma104 cells with ouabain increased MRP1-mRNA expression and altered the localization of MRP1 in these cells. Our results suggest that some cells may have mechanisms to protect themselves from ouabain toxicity and that MRP1 may have a role in controlling the toxic effects of ouabain. PMID- 11410340 TI - Study of the mode of action of endopolygalacturonase from Fusarium moniliforme. AB - One endopolygalacturonase from Fusarium moniliforme was purified from the culture broth of a transformed strain of Saccharomyces cerevisiae. Its kinetic parameters and mode of action were studied on galacturonic acid oligomers and homogalacturonan. The dimer was not a substrate for the enzyme. The enzyme was shown to follow Michaelis-Menten behaviour towards the other substrates tested. Affinity and maximum rate of hydrolysis increased with increasing chain length, up to the hexamer or heptamer, for which V(max) was in the same range as with homogalacturonan. The enzyme was demonstrated to have a multi-chain attack mode of action and its active site included five subsites ranging from -3 to +2. The final products of hydrolysis of homogalacturonan were the monomer and the dimer of galacturonic acid. PMID- 11410341 TI - Molecular phylogeny of the genus Bartonella: what is the current knowledge? AB - Species of the genus Bartonella are involved in an increasing variety of human diseases. In addition to the 14 currently recognized species, several Bartonella strains have been recovered from a wide range of wild and domestic mammals in Europe and America. Such a high diversity of geographic distributions, animal reservoirs, arthropod vectors and pathogenic properties makes clarification of our knowledge about the phylogeny of Bartonella species necessary. Phylogenetic data have been inferred mainly from 16S rDNA, 16S--23S rRNA intergenic spacer, citrate synthase and 60 kDa heat-shock protein gene sequences, which are available in GenBank. Comparison of phylogenetic organizations obtained from various genes allowed six statistically significant evolutionary clusters to be identified. Bartonella bacilliformis and Bartonella clarridgeiae appear to be divergent species. Bartonella henselae, Bartonella koehlerae and Bartonella quintana cluster together, as well as Bartonella vinsonii subsp. vinsonii and B. vinsonii subsp. berkhoffii. The fifth group includes bacteria isolated from various rodents that belong to native species from the New World and in the sixth, Bartonella tribocorum, Bartonella elizabethae and Bartonella grahamii are grouped with several strains associated with Old World indigenous rodents. The position of the other species could not be consistently determined. As some cat- or rodent-associated Bartonella appeared to cluster together, it has been suggested that these bacteria and their reservoir hosts may co-evolve. Lack of host specificity, however, seems to be frequent and may reflect the influence of vector specificity. Host or vector specificity may also explain the current geographic distribution of Bartonella species. PMID- 11410342 TI - PhaR, a protein of unknown function conserved among short-chain-length polyhydroxyalkanoic acids producing bacteria, is a DNA-binding protein and represses Paracoccus denitrificans phaP expression in vitro. AB - A putative regulatory protein, PhaR, which was identified in the polyhydroxyalkanoic acid synthetic locus (phaZCPR) in Paracoccus denitrificans, was investigated. The PhaR protein purified from a recombinant Escherichia coli was estimated to be 22 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, being consistent with the mass calculated from the nucleotide sequence. The molecular mass was determined to be 93 kDa by size-exclusion chromatography, suggesting that the protein formed a tetramer. A gel mobility shift assay showed that PhaR specifically bound to the intergenic region of phaC- phaP. In a cell-free protein synthesis system using E. coli S30 extract, the expression of the phaP gene was repressed by the addition of purified PhaR. These results suggest that PhaR is a DNA-binding protein and may play a role in the regulation of phaP gene expression. PMID- 11410343 TI - Effects of 2,2',5,5'-tetrachlorobiphenyl and biphenyl on cell membranes of Ralstonia eutropha H850. AB - The effects of 2,2',5,5'-tetrachlorobiphenyl (TeCB), a PCB congener, and biphenyl on the cytoplasmic membranes of Ralstonia eutropha H850 were investigated by measuring fluorescence polarization using 1,6-diphenyl-1,3,5-hexatriene (DPH) as the probe, and determining the cellular fatty acid compositions. TeCB significantly affected the membrane of R. eutropha H850 cells grown on fructose by decreasing DPH fluorescence polarization. In contrast, the membrane of cells grown on biphenyl showed a considerably less significant effect of TeCB on membrane polarization than in fructose-grown cells. An increase in the ratio of total saturated to unsaturated fatty acids in cells grown on biphenyl suggested less of a fluidizing effect of TeCB on membranes in those cells. When biphenyl grown cells were transferred back to a fructose medium, they required 25 generations for the membrane polarization and fatty acid compositions of these cells to revert back to those of the initial fructose-grown cells. The re adaptation to a change in temperature required only five generations to return to normal. These results show that biphenyl affects cells in more ways than simply fluidizing the cytoplasmic membrane. PMID- 11410344 TI - Enhancement of minor laccases production in the basidiomycete Marasmius quercophilus C30. AB - The white-rot fungus Marasmius quercophilus C30 is able to produce several laccases. The proportion of the enzymes produced depends on culture conditions. On malt medium, LAC1 was produced continuously over the 14 days of the cultivation period and was the only activity detectable. Copper increased total laccase activity by a factor 10 and induced the transient expression of one or more extra laccases in the culture medium. A combination of copper and p hydroxybenzoic acid made it possible to extend the expression of induced laccase activities over the cultivation period and to reach a maximum activity 30 times higher than in non-induced culture. Extracellular laccases produced in this last condition were eluted as four peaks on an anion exchange column and were partially characterized. PMID- 11410345 TI - Gene re-arrangement and fusion in Mycoplasma gallisepticum thyA-nrdFEI locus. AB - Sequencing of Mycoplasma gallisepticum genome fragment containing thymidylate synthase and ribonucleotide reductase gene clusters reveals both its unusual organization and gene content. Sequence analysis indicates the presence of a gene whose product can be considered as a fusion of two full size proteins: the N terminal part shows significant similarity to mycoplasmal dihydrofolate reductases, while the C-terminal part of the polypeptide chain shows significant similarity to eukaryotic deoxycytidylate deaminase. Phylogenetic analysis has suggested that the C-terminal part of the M. gallisepticum fusion gene and eukaryotic deoxycytidylate deaminase genes are xenologous. No chromosomal regions encoding peptides similar to the C-terminal part of this fusion protein were found in completely sequenced genomes of Mycoplasma genitalium and Mycoplasma pneumoniae. Genes for ribonucleoside diphosphate reductase alpha chain (nrdE), NrdI protein (nrdI), and ribonucleoside diphosphate reductase beta chain (nrdF) have an opposite direction of transcription with respect to genes for thymidylate synthase (thyA), and dihydrofolate reductase-deoxycytidylate deaminase fusion protein. PMID- 11410346 TI - Sensitive genotyping of Cryptosporidium parvum by PCR-RFLP analysis of the 70 kilodalton heat shock protein (HSP70) gene. AB - A polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis of a 587-bp region of the Cryptosporidium parvum 70-kDa heat shock protein (HSP70) gene was developed for the detection and discrimination of the two major genotypes of C. parvum, genotype 1 and genotype 2. Ten Cryptosporidium isolates from non-immunocompromised people were identified as genotypes 1 and 2 (five each) by DNA sequencing of the 587-bp PCR product. This distinction was also achieved with the combination of two endonucleases, HinfI and ScaI, which generated a specific pattern for each genotype. A thorough screening of published sequences showed that this combination of enzymes could also be used for the discrimination of other species/genotypes of Cryptosporidium, especially Cryptosporidium meleagridis and the 'dog' genotype of C. parvum, both of which are infectious in humans. The PCR, conducted on genotypes 1 and 2 of C. parvum, could detect one oocyst per reaction. This new and sensitive genotyping procedure should be of particular interest when applied to the monitoring of water resources in which low concentrations of parasites usually occur. PMID- 11410347 TI - Characterisation of a cyclophilin isoform in Trypanosoma cruzi. AB - The immunosuppressive drug cyclosporin A (CsA) has shown antiparasitic activity against several protozoans and helminths, when complexed to proteins called cyclophilins (CyPs). In this paper, the molecular characterisation of one member of the CyP family in Trypanosoma cruzi is reported. TcCyP19 gene proved to be highly conserved compared to CyPs from other organisms and was highly homologous to a Trypanosoma brucei brucei CyPA. This gene was expressed in Escherichia coli and the purified recombinant protein exhibited a peptidyl prolyl cis-trans isomerase activity that was inhibited by CsA (IC(50) = 18.4 + /-0.8 nM). The TcCyP19 gene was located on two chromosomal bands in T. cruzi CL Brener clone. PMID- 11410348 TI - Protein expression under uracil privation in Lactobacillus sakei. AB - Lactobacillus sakei is a lactic acid bacterium belonging to the natural flora of fresh and vacuum-packed meat, and is used as the starter for manufacturing fermented sausages. This species is now being studied at the genetic level. We investigated uracil prototrophy of strain 23K in order to validate the proteomic approach to study metabolism regulations. Cells grown without uracil had lower growth rates than with uracil. Protein analysis by 2D gel electrophoresis showed that at least three polypeptides were specifically induced in the absence of uracil. Two of these polypeptides were identified as orotate phosphoribosyl transferase, catalyzing the fifth step of pyrimidine biosynthesis, and PyrR, the transcriptional regulator of the pyr operon, respectively. PMID- 11410349 TI - Interaction of Knr4 protein, a protein involved in cell wall synthesis, with tyrosine tRNA synthetase encoded by TYS1 in Saccharomyces cerevisiae. AB - The Knr4 protein, known to be involved in the regulation of cell wall assembly in Saccharomyces cerevisiae, strongly interacts with the tyrosine tRNA synthetase protein encoded by TYS1 as demonstrated by the genetic two-hybrid system and a biochemical pull-down experiment using GST--Tys1p fusion. Data reported here raise the possibility that this physical interaction between these proteins is required for dityrosine formation during the sporulation process. In addition, it is shown that the efficiency of spores formation was drastically reduced in diploid cells homozygous for the disruption of KNR4 or for a temperature sensitive mutation of TYS1, although this effect could be independent of their protein interaction. Altogether, these data provide novel functions of Knr4p and Tys1p to those that were known before. PMID- 11410350 TI - Construction and characterization of a Xanthomonas oryzae pv. oryzae bacterial artificial chromosome library. AB - Xanthomonas oryzae pv. oryzae is an important plant pathogen which causes bacterial blight of rice. To facilitate genome studies of this bacterium, we have constructed a bacterial artificial chromosome (BAC) library of strain MAFF 311018. It consisted of 750 clones representing 16 genome equivalents, and had an insert size ranging from 20 to 220 kb with an average size of 107 kb. This library is the first to be constructed from a X. oryzae pv. oryzae strain. The usefulness of this library was demonstrated through polymerase chain reaction screening of 11 genes and the 16S--23S rDNA spacer region in a 192-clone subset, representing five genome equivalents. The results obtained showed an average of 5.9 BAC clones per screening. This result is in good agreement with the estimated size of the test library, indicating that the constructed BAC library can be used to facilitate genome analysis of X. oryzae pv. oryzae. PMID- 11410351 TI - Phylogenetic analysis of archaeal 16S rRNA libraries from the rumen suggests the existence of a novel group of archaea not associated with known methanogens. AB - Molecular diversity of rumen archaea was analyzed by PCR amplification and sequencing of two 16S rRNA clone libraries prepared from the bovine rumen fluid using two different archaea-specific primer sets. The first library of 19 clones which was generated with primers D30 and D33, produced essentially two groups of sequences, one affiliated with Methanomicrobium mobile (21% of clones) and the other -- with the uncultured archaeal sequences from anaerobic digester, which are distantly associated with Thermoplasma (79% of clones). The second library of 25 clones, which was generated with primers 0025e Forward and 1492 Reverse, produced a higher degree of diversity: in addition to the previous two groups, with the M. mobile- (56%) and Thermoplasma-associated sequences (20%), four clones (16%) were identified as Methanobrevibacter spp. The remaining two sequences were associated with unidentified archaeal sequences from the rumen and swine waste. Phylogenetic placement of eight almost complete 16S rRNA sequences revealed the existence of a novel cluster of the rumen Euryarchaeota, which is not affiliated with the known methanogenic archaea. PMID- 11410352 TI - Dual control of hydrogen cyanide biosynthesis by the global activator GacA in Pseudomonas aeruginosa PAO1. AB - The global response regulator GacA of Pseudomonas aeruginosa PAO1 positively controls the production of the quorum sensing signal molecule N-butanoyl homoserine-lactone (C4-HSL) and hence the synthesis of several C4-HSL-dependent virulence factors, including hydrogen cyanide (HCN). This study presents evidence that GacA positively influences the transcription of the rhlI gene, specifying C4 HSL synthase, explaining the quorum sensing-dependent transcriptional control of the HCN biosynthetic genes (hcnABC). In addition, GacA was found to modulate hcn gene expression positively at a post-transcriptional level involving the hcnA ribosome-binding site. Thus, the activating effect of GacA on cyanogenesis results from both transcriptional and post-transcriptional mechanisms. PMID- 11410353 TI - On the origin of Ser/Thr kinases in a prokaryote. AB - The family of Ser/Thr and/or Tyr kinases and that of His kinases play essential roles in signal transduction. For a long time, the former has been found in eukaryotes, the latter in prokaryotes. Studies in the last decade have shown, however, that most bacteria possess from one to more than 10 genes encoding Ser/Thr kinases. This observation raises an important question concerning the evolutionary origin of Ser/Thr kinases found in bacteria. To answer this question, we have analyzed a family of 11 genes encoding Ser/Thr kinases in the cyanobacterium Synechocystis sp. PCC 6803. This bacterium contains the largest number of Ser/Thr kinases among all bacteria whose genomic sequences have been released so far. In this study, we have developed a user-friendly computer program for statistical analysis of codon usages and GC content. The results demonstrate that Ser/Thr kinases have similar codon usages and GC contents as the average of all possible open reading frames (ORFs) deduced from the genome. In contrast, ORFs encoding transposases, as a control in our analysis, display a disparity in both codon usage and GC content, confirming their multiple origin and genetic promiscuity. In light of our results, we propose that Ser/Thr kinases existed before the divergence between prokaryotes and eukaryotes during evolution, or were laterally transferred into prokaryotes at the early stages of bacterial evolution. If Ser/Thr kinases have persisted ever since in prokaryotes under evolutionary pressure, it is then expected that they play important, possibly even essential roles in regulating bacterial activities as do their counterparts in eukaryotes. PMID- 11410354 TI - Insertional mutagenesis of an industrial strain of Streptococcus thermophilus. AB - Random mutagenesis of an industrial strain of Streptococcus thermophilus was achieved through an adapted version of a two-plasmid system. The mutagenesis strategy is based on random integration of derivatives of the non-replicative (Rep(-)) plasmid pORI19 by means of homologous recombination following a temperature shift that eliminates replication of the temperature-sensitive (Rep(ts)) helper plasmid pVE6007. In this way mutants were generated which were affected in bacteriophage sensitivity or sucrose metabolism. Homologues were identified of a protein related to folate metabolism from a bacteriophage resistant mutant and of two subunits of an oligopeptide transport system from a mutant deficient in sucrose utilisation. PMID- 11410355 TI - The LlaGI restriction and modification system of Lactococcus lactis W10 consists of only one single polypeptide. AB - The naturally occurring 12.1-kb plasmid, pEW104, in Lactococcus lactis ssp. cremoris W10 was found to confer decreased bacteriophage sensitivity to its host. Plasmid pEW104 encodes a non-classic restriction and modification (R/M) system, named LlaGI, consisting of only one single polypeptide. Analysis of the amino acid sequence revealed the presence of a catalytic motif and seven helicase-like motifs (DEAD-box motifs) characteristic of type I and III endonucleases, followed by four conserved methylase motifs characteristic of adenine-methylases. A comparison between LlaGI and the very similar R/M system, LlaBIII, suggests that the C-terminal region of LlaGI, apparently containing no known motifs, could possibly specify target DNA recognition. Conceivably, the LlaGI gene is included in the operon of the plasmid replication machinery. Finally, it is proposed that LlaGI represents a variant of the type I R/M systems. PMID- 11410356 TI - Independent replication of the plasmids pRN1 and pRN2 in the archaeon Sulfolobus islandicus. AB - The 5.4-kb and 6.9-kb plasmids pRN1 and pRN2 from the crenarchaeon Sulfolobus islandicus are name-giving for a small family of archaeal plasmids. Both plasmids have hitherto been supposed to be dependent on each other because they are always found together in their natural host. Here we demonstrate that each of the plasmids can stably propagate and replicate on its own independent of the other plasmid. Moreover, we could show that in vivo the plasmids bear tightly bound proteins. PMID- 11410357 TI - Bacterial community analysis of Indonesian hot springs. AB - We report the first attempts to describe thermophilic bacterial communities in Indonesia's thermal springs using molecular phylogenetic analyses. 16S rRNA genes from laboratory cultures and DNA directly amplified from three hot springs in West Java were sequenced. The 22 sequences obtained were assignable to the taxa Proteobacteria, Bacillus and Flavobacterium, including a number of clades not normally associated with thermophily. PMID- 11410358 TI - Unusual adaptive, cross protection responses and growth phase resistance against peroxide killing in a bacterial shrimp pathogen, Vibrio harveyi. AB - Oxidant induced protection against peroxide killing was investigated in a prawn bacterial pathogen, Vibrio harveyi. Exposure to 250 microM H(2)O(2) induced adaptive protection against subsequent exposure to killing concentrations of H(2)O(2). In addition, 200 microM t-butyl hydroperoxide (tBOOH) induced cross protection to H(2)O(2) killing. On the other hand, peroxide pretreatment did not induce protection against tBOOH killing. Peroxide induced adaptive and cross protection responses required new protein synthesis and were abolished by addition of a protein synthesis inhibitor. Pretreatments of V. harveyi with 250 microM H(2)O(2) and 200 microM tBOOH induced an increase in peroxide scavenging enzymes, catalase and alkyl hydroperoxide reductase subunit C. In addition, stationary phase cells of V. harveyi were more resistant to H(2)O(2) and iodoacetamide killing but highly susceptible to tBOOH killing compared to exponential phase cells. Many aspects of the oxidative stress response of V. harveyi are different from those of other bacteria and these factors may be important for bacterial survival in the environment and during interactions with host shrimp. PMID- 11410359 TI - Interactions of Escherichia coli strains of non-EPEC serogroups that carry eae and lack the EAF and stx gene sequences with undifferentiated and differentiated intestinal human Caco-2 cells. AB - Escherichia coli strains of non-EPEC serotypes that carry eae and lack the EAF and the Shiga toxin (stx) gene sequences have been found in acute diarrhea. Both the cell association and the cell entry of these strains in human intestinal epithelial cells were studied as a function of cell differentiation and polarization. The eae+/EAF-/stx- non-EPEC E. coli strains invaded undifferentiated Caco-2 cells more efficiently than differentiated cells. In contrast, prototype EPEC strain E2348/69 did not show significative differences from invasion rates of undifferentiated and differentiated cells. The uptake of these strains was greatly enhanced by pretreatment of differentiated Caco-2 cells with EGTA. These results suggest that the eae+/EAF-/stx- non-EPEC E. coli invasion of intestinal cells may be dependent on receptors expressed on the surface of undifferentiated cells and the basolateral pole of differentiated cells. PMID- 11410360 TI - Histone H1 diversity: bridging regulatory signals to linker histone function. AB - Genes encoding linker histone variants have evolved to link their expression to signals controlling the proliferative capacities of cells, i.e. cycling and growth-arrested cells express distinct and specific H1 subtypes. In metazoan, these variants show a tripartite structure, with considerably divergent sequences in their amino and carboxyl terminus domains. The aim of this review is to show how specific regulatory signals control the expression of an individual H1 and to discuss the functional significance of the two variables associated with a linker histone: its primary sequence and the timing of its expression. PMID- 11410361 TI - Degenerate oligonucleotide gene shuffling (DOGS): a method for enhancing the frequency of recombination with family shuffling. AB - Improvement of the biochemical characteristics of enzymes has been aided by misincorporation mutagenesis and DNA shuffling. Shuffling techniques can be used on a collection of mutants of the same gene, or related families of genes can be shuffled to produce mutants encoding chimeric gene products. One difficulty with current shuffling procedures is the predominance of unshuffled ("parental") molecules in the pool of mutants. We describe a procedure for gene shuffling using degenerate primers that allows control of the relative levels of recombination between the genes that are shuffled and reduces the regeneration of unshuffled parental genes. This procedure has the advantage of avoiding the use of endonucleases for gene fragmentation prior to shuffling and allows the use of random mutagenesis of selected segments of the gene as part of the procedure. We illustrate the use of the technique with a diverse family of beta-xylanase genes that possess widely different G+C contents. PMID- 11410362 TI - Cloning and molecular characterization of a novel gene encoding a WD-repeat protein expressed in restricted areas of adult rat brain. AB - We describe the cloning and characterization of a novel rat gene, bwd (brain enriched WD), which encodes a protein with four copies of the WD amino acid motif, suggesting involvement in protein-protein interaction and a regulatory function in the cell. Northern analysis reveals two size classes of mRNA (1.8 and 2.2 kb), expressed in many adult tissues and developmental stages. Expression is highest in brain, where the longer of the two RNAs predominates. cDNA sequences show that both RNAs encode the identical protein, differing only in their 3' untranslated regions, where the longer transcript contains two RNA instability signals (AUUUA). In situ hybridization to bwd RNA in adult brain shows a highly restricted pattern, localizing expression mainly to the Purkinje and granule neurons of the cerebellum, the pyramidal cells of the hippocampus, and the dentate gyrus. In cryosections of rat cerebellum and kidney, BWD is shown by immunohistochemistry to be localized in the nucleus and cytoplasm of cerebellar Purkinje and granule neurons, and in predominantly the cytoplasm of cells surrounding kidney ducts. Taken together, these results suggest a specialized function for BWD in the brain. Sequence similarity comparisons with bwd reveal structural homologs of unknown function in human, mouse, Drosophila, Arabidopsis and C. elegans, and provide evidence that this set of sequences forms a new subfamily of WD-repeat genes. By sequence comparisons with expressed sequence tags (ESTs), the human homolog of bwd is predicted to reside in the chromosome 1q12-23 region, where several genetic diseases are known to map. PMID- 11410363 TI - Total tetra knockout of GP138 multigene family implicated in cell interactions in Dictyostelium discoideum. AB - The cellular slime mold Dictyostelium discoideum reproduces sexually under submerged and dark conditions. A cell surface glycoprotein gp138 has been identified as a target molecule for cell fusion-blocking antibodies, and is considered to be indispensable for the sexual cell fusion in this organism. Currently, four isoforms of gp138, DdFRP1alpha, DdFRP1beta, DdFRP2, and DdFRP3, are known. Genes encoding the latter three isoforms, GP138C, GP138A, and GP138B, have been isolated, comprising a GP138 multigene family. Here we isolated the fourth GP138 gene, GP138D, encoding DdFRP1alpha. These GP138 genes were found to cluster in a tandem array on chromosome 5, being bordered by two GP138-like sequences highly homologous to them but truncated. To clarify functional relationships among the GP138 family members, the entire GP138 region was deleted by a single knockout. Northern hybridization and western immuno-blotting analyses confirmed complete losses of GP138 mRNA and DdFRPs in the knockout strains, indicating that there are no more GP138 genes. Unexpectedly, however, the GP138 null mutants were fully potent for both sexual cell fusion and subsequent development. In addition, the original fusion-blocking antibodies detected a cell surface protein of close electrophoretic mobility to gp138 in the knockouts, suggesting the possibility that the actual target molecule of the fusion-blocking antibodies was not DdFRPs but this unidentified component. Since GP138-null mutants exhibited no obvious defects either in growth or asexual development, the real function of the GP138 family is unknown. Nevertheless, the expression levels of other developmental genes such as acaA, csaA, cotA-C, and spiA appeared to be altered in the GP138-null mutants. Therefore, it seems to have a non-critical but some role(s) during asexual development. PMID- 11410364 TI - Intron loss in the SART1 genes of Fugu rubripes and Tetraodon nigroviridis. AB - The human SART1 gene was initially identified in a screen for proteins recognised by IgE, which may be implicated in atopic disease. We have examined the genomic structure and cDNA sequence of the SART1 gene in the compact genomes of the pufferfish Fugu rubripes and Tetraodon nigroviridis. The entire coding regions of both the Fugu and Tetraodon SART1 genes are contained within single exons. The Fugu gene contains only one intron located in the 5' untranslated region. Southern blot hybridisation of Fugu genomic DNA confirmed the SART1 gene to be single copy. Partial genomic structures were also determined for the human, mouse, Drosophila and C. elegans SART1 homologues. The human and mouse genes both contain many introns in the coding region, the human gene possessing at least 20 exons. The Drosophila and C. elegans homologues contain 6 and 12 exons, respectively. This is only the second time such a difference in the organization of homologous Fugu and human genes has been reported. The Fugu and Tetraodon SART1 genes encode putative proteins of 772 and 774 aa, respectively, each having 65% amino acid identity to human SART1. Leucine zipper and basic motifs are conserved in the predicted Fugu and Tetraodon proteins. PMID- 11410365 TI - Sequence and analysis of the murine Hmgiy (Hmga1) gene locus. AB - The HMGIY non-histone proteins play important roles as architectural transcription factors that regulate gene transcription in mammalian cells and also act as host-supplied cofactors necessary for retroviral integration. The genes coding for the HMGIY proteins are proto-oncogenes, and their aberrant or over-expression is correlated with both neoplastic transformation and metastatic progression in a wide variety of tumors. Here, we report the first complete sequence of the murine Hmgiy (a.k.a. Hmga1) gene and provide a detailed comparison of this with the sequence and organization of the human HMGIY gene, including an analysis of its promoter region with the previously unreported 5' upstream region of the human gene. These analyses reveal a remarkable degree of overall sequence conservation in both the protein coding and promoter regions of the murine and human genes, including conservation of the c-Myc binding site that has been demonstrated to regulate murine Hmgiy transcription (Wood et al., 2000. Mol. Cell. Biol. 20, 5490-5502). The promoters of both genes contain other conserved transcription factor binding sites that may also represent important cis-regulatory elements. Two exons present in the 5' untranslated region of the human gene, however, are missing from the murine gene, suggesting that these two closely related mammalian species regulate transcription of their Hmgiy genes in an individualistic manner. PMID- 11410366 TI - Sequence, chromosomal location and expression analysis of the murine homologue of human RAD51L2/RAD51C. AB - The Rad51 protein has been shown to play a vital role in the DNA repair process. In humans, its interaction with proteins like BRCA1 and BRCA2 has provided an insight into the mechanism of how these molecules function as tumor suppressors. Several members of the Rad51-like family have been recently identified, including RAD51L2. This gene has been found to be amplified in breast tumors suggesting its role in tumor progression. Here, we describe the cloning of the murine homologue of the human RAD51L2/RAD51C gene. Sequence analysis has revealed that the murine Rad51l2 protein is 86% identical and 93% similar to its human homologue. In spite of such high sequence conservation, the murine protein lacks the first nine amino acids present in the human protein. We have cloned and confirmed the sequence of the 5' end of the murine Rad51l2 cDNA using 5' RACE technique as well as by sequencing the genomic region flanking the first exon of the murine Rad51l2 gene. Northern analysis shows that Rad51l2 is expressed in several adult tissues as well as in embryos at various developmental stages. The murine Rad51l2 gene maps to chromosome 11 and is located in the syntenic region of human chromosome 17q22 23, where the human RAD51L2 is present. PMID- 11410367 TI - Gene structure and expression of phospholemman in mouse. AB - Phospholemman (PLM) is a small transmembrane cardiac protein that is the major sarcolemmal substrate for phosphorylation in response to adrenergic stimulation. PLM likely plays a role in muscle contractility and cell volume regulation through its function as a channel or a channel regulator. We are the first to describe the structure of the PLM gene and to demonstrate PLM cDNA splice variants. We cloned the murine PLM cDNA and used it as a probe to isolate the gene from a 129/SvJ genomic library. The gene contains seven introns and eight exons. The coding sequence is interrupted by five introns; the 5' untranslated region by two. Using rapid amplification of 5' cDNA ends we identified transcription start sites and four splice variants of the 5' untranslated domain. There was no TATA box or CAAT box in the putative promoter regions. The gene has several stretches of dinucleotide repeats. The 3' untranslated domains of mouse PLM cDNA clones show sequence differences not accounted for by alternative splicing. Mouse PLM shares 93, 83 and 80% amino acid identity with rat, dog, and human PLMs, respectively. Tissue expression of murine PLM parallels that in other species, being highest in heart, skeletal muscle, and liver. PMID- 11410368 TI - Molecular characterization of homologues of both subunits A (SPO11) and B of the archaebacterial topoisomerase 6 in plants. AB - The Spo11 protein is an eukaryotic homologue of the topoisomerase 6 subunit A from archaebacteria. In yeast Spo11p has been found to bind covalently to double strand breaks (DSBs) during meiosis. Single homologues of the SPO11 gene exist in various eukaryotes, except plants. Previously, we found in the Arabidopsis thaliana genome two ancient paralogs, AtSPO11-1 and 2. Here we report on the molecular characterization of a third one, AtSPO11-3. This puzzling finding might be explained by the fact that we detected additionally--for the first time outside of the archaebacterial kingdom--a homologue of the subunit B of topoisomerase 6, AtTOP6B. Both AtSPO11-3 and AtTOP6B are abundantly expressed in Arabidopsis and EST comparisons indicate the presence of both genes in various plant species. Via two hybrid studies we could demonstrate that full length AtTop6B is able to interact with AtSpo11-2 and 3 but not with AtSpo11-1. Our data suggest that plants possess in contrast to other eukaryotes an additional archaebacterial kind of topoisomerase. PMID- 11410369 TI - Structure of the human M(2) muscarinic acetylcholine receptor gene and its promoter. AB - The M(2) muscarinic receptor inhibits the release of acetylcholine from cholinergic fibers in the lungs and elsewhere. In airway parasympathetic neurons, M(2) receptor expression is decreased by viral infections and by interferon gamma, increasing actylcholine release. Dexamethasone increases M(2) receptor expression, decreasing acetylcholine release. We carried out 5' rapid amplification of cDNA ends beginning with mRNA from human heart and IMR32 human neuroblastoma cells. This demonstrated a 5' UTR of 100 BP, corresponding to two sequences on chromosome 7, separated by a 22.6 kB intron. The splice acceptor site is at -45 relative to the initiating atg. The 3000 BP upstream of 5' RACE product were subcloned into a pGL3 luciferase reporter vector. Deletional constructs were expressed in IMR32 cells. These demonstrated that 412 BP provided full expression of the reporter gene, and suggested a repressor element between 1848 and -1510. PMID- 11410370 TI - Human-mouse comparative sequence analysis of the NEMO gene reveals an alternative promoter within the neighboring G6PD gene. AB - NEMO (NFkappaB essential modulator) is a non-catalytic subunit of the cytokine dependent IkappaB kinase complex that is involved in activation of the transcription factor NFkappaB. The human NEMO gene maps to Xq28 and is arranged head to head with the proximal G6PD gene. Mutations in NEMO have recently been associated with Incontinentia Pigmenti (Smahi et al., Nature 405 (2000) 466), an X-linked dominant disorder. Three alternative transcripts with different non coding 5' exons (1a, 1b and 1c) of NEMO have been described. In order to identify regulatory elements that control alternative transcription we have established the complete genomic sequence of the murine orthologs Nemo and G6pdx. Sequence comparison suggests the presence of two alternative promoters for NEMO/Nemo. First, a CpG island is shared by both genes driving expression of the NEMO/Nemo transcripts containing exons 1b and 1c in one direction and the housekeeping gene G6PD/G6pdx in the opposite direction. In contrast to human, an additional variant of exon 1c, named 1c+, was identified in several tissues of the mouse. This larger exon utilizes an alternative donor site located 1594 bp within intron 1c. The putative second promoter for NEMO/Nemo transcripts starting with exon 1a is unidirectional, and not associated with a CpG island. Surprisingly, this promoter is located in the second intron of G6PD/G6pdx. It shows very low basal activity and may be involved in stress/time- and/or tissue-dependent expression of NEMO. To our knowledge, an overlapping gene order similar to the G6PD/NEMO complex has not been described before. PMID- 11410371 TI - Cloning and characterization of two Arabidopsis genes that belong to the RAD21/REC8 family of chromosome cohesin proteins. AB - Sister chromatid cohesion is required for proper chromosome segregation during cell division. One group of proteins that is essential for sister chromatid cohesion during mitosis and meiosis is the RAD21/REC8 family of cohesin proteins. Two cohesin proteins are found in yeast; one that functions mainly in mitosis while the other participates in meiosis. In contrast, only one cohesin gene appears to be present in Drosophila. In previous studies we identified an Arabidopsis cohesin protein that is required for meiosis. In this report we describe the isolation and characterization of two additional Arabidopsis cohesin genes. The structure of the genes suggests that they arose via a gene duplication event followed by extensive sequence evolution. Transcripts for the two genes are present throughout the plant and are highest in regions of active cell division, suggesting that the proteins may participate in chromosome cohesion during mitosis. PMID- 11410372 TI - Differential gene expression in fully-grown oocytes between gynogenetic and gonochoristic crucian carps. AB - Silver crucian carp (Carassius auratus gibelio) is a unique triploid bisexual species that can reproduce by gynogenesis. As all other gynogenetic animals, it keeps its chromosome integrity by inhibiting the first meiosis division (no extrusion of the first pole body). To understand the molecular events governing this reproduction mode, suppression subtractive hybridization was used to identify the genes differentially expressed in fully-grown oocytes of the gynogenetic and gonochoristic crucian carp (gyno-carp and gono-carp). From two specific subtractive cDNA libraries, the clones screened out by dot blots and virtual Northern blots were chosen to clone full-length cDNA by RACE. Four differentially expressed genes were obtained. Two are novel genes and are expressed specifically in the oocytes. The gyno-carp stores much more mRNA of cyclin A2, a new member of the fish A-type cyclin gene, in its fully-grown oocyte than in the gono-carp. The last gene is histone H2A. The histone H2As of these two closely related crucian carps are quite different in the C-terminus. Preliminary characterization of the four genes has been analyzed by nucleotide and deduced amino acid sequence and Northern analysis. PMID- 11410373 TI - Canonical binding arrays as molecular recognition elements in the immune system: tetrahedral anions and the ester hydrolysis transition state. AB - The structures, obtained by X-ray crystallography, of the binding sites of catalytic antibodies raised to bind different phosphonates are compared. Although the amino acid sequences differ, all exhibit a tetrahedral array of hydrogen bond donors (a 'canonical binding array') complementary to the tetrahedral anion, which represents a 'transition state epitope' for the basic hydrolysis of esters and amides. Antibodies for phosphates, arsonates, and sulfonates are found also to possess the tetrahedral anion canonical binding array. PMID- 11410374 TI - Construction of desosamine containing polyketide libraries using a glycosyltransferase with broad substrate specificity. AB - BACKGROUND: Combinatorial biosynthesis techniques using polyketide synthases (PKSs) in heterologous host organisms have enabled the production of macrolide aglycone libraries in which many positions of the macrolactone ring have been manipulated. However, the deoxysugar moieties of macrolides, absent in previous libraries, play a critical role in contributing to the antimicrobial properties exhibited by compounds such as erythromycin. Since the glycosidic components of polyketides dramatically alter their molecular binding properties, it would be useful to develop general expression hosts and vectors for synthesis and attachment of deoxysugars to expand the nature and size of such polyketide libraries. RESULTS: A set of nine deoxysugar biosynthetic and auxiliary genes from the picromycin/methymycin (pik) cluster was integrated in the chromosome of Streptomyces lividans to create a host which synthesizes TDP-D-desosamine. The pik desosaminyl transferase was also included so that when the strain was transformed with a previously constructed library of expression plasmids encoding genetically modified PKSs that produce different macrolactones, the resulting strains produced desosaminylated derivatives. Although conversion of the macrolactones was generally low, bioassays revealed that, unlike their aglycone precursors, these novel macrolides possessed antibiotic activity. CONCLUSIONS: Based on the structural differences among the compounds that were glycosylated it appears that the desosaminyl transferase from the pik gene cluster is quite tolerant of changes in the macrolactone substrate. Since others have demonstrated tolerance towards modifications in the sugar substituent, one can imagine employing this approach to alter both polyketide and deoxysugar pathways to produce 'unnatural' natural product libraries. PMID- 11410375 TI - Two sequence elements of glycosyltransferases involved in urdamycin biosynthesis are responsible for substrate specificity and enzymatic activity. AB - BACKGROUND: Two deoxysugar glycosyltransferases (GTs), UrdGT1b and UrdGT1c, involved in urdamycin biosynthesis share 91% identical amino acids. However, the two GTs show different specificities for both nucleotide sugar and acceptor substrate. Generally, it is proposed that GTs are two-domain proteins with a nucleotide binding domain and an acceptor substrate site with the catalytic center in an interface cleft between these domains. Our work aimed at finding out the region responsible for determination of substrate specificities of these two urdamycin GTs. RESULTS: A series of 10 chimeric GT genes were constructed consisting of differently sized and positioned portions of urdGT1b and urdGT1c. Gene expression experiments in host strains Streptomyces fradiae Ax and XTC show that nine of 10 chimeric GTs are still functional, with either UrdGT1b- or UrdGT1c-like activity. A 31 amino acid region (aa 52-82) located close to the N terminus of these enzymes, which differs in 18 residues, was identified to control both sugar donor and acceptor substrate specificity. Only one chimeric gene product of the 10 was not functional. Targeted stepwise alterations of glycine 226 (G226R, G226S, G226SR) were made to reintroduce residues conserved among streptomycete GTs. Alterations G226S and G226R restored a weak activity, whereas G226SR showed an activity comparable with other functional chimeras. CONCLUSIONS: A nucleotide sugar binding motif is present in the C-terminal moiety of UrdGT1b and UrdGT1c from S. fradiae. We could demonstrate that it is an N terminal section that determines specificity for the nucleotide sugar and also the acceptor substrate. This finding directs the way towards engineering this class of streptomycete enzymes for antibiotic derivatization applications. Amino acids 226 and 227, located outside the putative substrate binding site, might be part of a larger protein structure, perhaps a solvent channel to the catalytic center. Therefore, they could play a role in substrate accessibility to it. PMID- 11410376 TI - Biosynthesis of the orthosomycin antibiotic avilamycin A: deductions from the molecular analysis of the avi biosynthetic gene cluster of Streptomyces viridochromogenes Tu57 and production of new antibiotics. AB - BACKGROUND: Streptomyces viridochromogenes Tu57 is the producer of avilamycin A. The antibiotic consists of a heptasaccharide side chain and a polyketide-derived dichloroisoeverninic acid as aglycone. Molecular cloning and characterization of the genes governing the avilamycin A biosynthesis is of major interest as this information might set the direction for the development of new antimicrobial agents. RESULTS: A 60-kb section of the S. viridochromogenes Tu57 chromosome containing genes involved in avilamycin biosynthesis was sequenced. Analysis of the DNA sequence revealed 54 open reading frames. Based on the putative function of the gene products a model for avilamycin biosynthesis is proposed. Inactivation of aviG4 and aviH, encoding a methyltransferase and a halogenase, respectively, prevented the mutant strains from producing the complete dichloroisoeverninic acid moiety resulting in the accumulation of new antibiotics named gavibamycins. CONCLUSIONS: The avilamycin A biosynthetic gene cluster represents an interesting system to study the formation and attachment of unusual deoxysugars. Several enzymes putatively responsible for specific steps of this pathway could be assigned. Two genes encoding enzymes involved in post-PKS tailoring reactions were deleted allowing the production of new analogues of avilamycin A. PMID- 11410377 TI - Towards a minimal motif for artificial transcriptional activators. AB - BACKGROUND: Most transcriptional activators minimally comprise two functional modules, one for DNA binding and the other for activation. Several activators also bear an oligomerization region and bind DNA as dimers or higher order oligomers. In a previous study we substituted these domains of a protein activator with synthetic counterparts [Mapp et al., Proc. Natl. Acad. Sci. USA 97 (2000) 3930-3935]. An artificial transcriptional activator, 4.2 kDa in size, comprised of a DNA binding hairpin polyamide tethered to a 20 residue activating peptide (AH) was shown to stimulate promoter specific transcription [Mapp et al., Proc. Natl. Acad. Sci. USA 97 (2000) 3930-3935]. The question arises as to the general nature and the versatility of this minimal activator motif and whether smaller ligands can be designed which maintain potent activation function. RESULTS: Here we have replaced the 20 amino acid AH peptide with eight or 16 residues derived from the activation domain of the potent viral activator VP16. The 16 residue activation module coupled to the polyamide activated transcription over two-fold better than the analogous AH conjugate. Altering the site of attachment of the activation module on the polyamide allowed reduction of the intervening linker from 36 atoms to eight without significant diminution of the activation potential. In this study we also exchanged the polyamide to target a different sequence without compromising the activation function further demonstrating the generality of this design. CONCLUSIONS: The polyamide activator conjugates described here represent a class of DNA binding ligands which are tethered to a second functional moiety, viz. an activation domain, that recruits elements of the endogenous transcriptional machinery. Our results define the minimal structural elements required to construct artificial, small molecule activators. If such activators are cell-permeable and can be targeted to designated sites in the genome, this series of conjugates may then serve as a tool to study mechanistic aspects of transcriptional regulation and eventually to modulate gene expression relevant to human diseases. PMID- 11410378 TI - Structure-based design and in-parallel synthesis of inhibitors of AmpC beta lactamase. AB - BACKGROUND: Group I beta-lactamases are a major cause of antibiotic resistance to beta-lactams such as penicillins and cephalosporins. These enzymes are only modestly affected by classic beta-lactam-based inhibitors, such as clavulanic acid. Conversely, small arylboronic acids inhibit these enzymes at sub-micromolar concentrations. Structural studies suggest these inhibitors bind to a well defined cleft in the group I beta-lactamase AmpC; this cleft binds the ubiquitous R1 side chain of beta-lactams. Intriguingly, much of this cleft is left unoccupied by the small arylboronic acids. RESULTS: To investigate if larger boronic acids might take advantage of this cleft, structure-guided in-parallel synthesis was used to explore new inhibitors of AmpC. Twenty-eight derivatives of the lead compound, 3-aminophenylboronic acid, led to an inhibitor with 80-fold better binding (2; K(i) 83 nM). Molecular docking suggested orientations for this compound in the R1 cleft. Based on the docking results, 12 derivatives of 2 were synthesized, leading to inhibitors with K(i) values of 60 nM and with improved solubility. Several of these inhibitors reversed the resistance of nosocomial Gram-positive bacteria, though they showed little activity against Gram-negative bacteria. The X-ray crystal structure of compound 2 in complex with AmpC was subsequently determined to 2.1 A resolution. The placement of the proximal two thirds of the inhibitor in the experimental structure corresponds with the docked structure, but a bond rotation leads to a distinctly different placement of the distal part of the inhibitor. In the experimental structure, the inhibitor interacts with conserved residues in the R1 cleft whose role in recognition has not been previously explored. CONCLUSIONS: Combining structure-based design with in-parallel synthesis allowed for the rapid exploration of inhibitor functionality in the R1 cleft of AmpC. The resulting inhibitors differ considerably from beta-lactams but nevertheless inhibit the enzyme well. The crystal structure of 2 (K(i) 83 nM) in complex with AmpC may guide exploration of a highly conserved, largely unexplored cleft, providing a template for further design against AmpC beta-lactamase. PMID- 11410379 TI - Physics and engineering: milestones in medicine. AB - The history of the development of the applications of physics and engineering in medicine provides an insight into contemporary practice and can help to mould the future. Physics and engineering form a continuum, and, in the present context, engineering is indistinguishable from applied physics. The modern scientific era, which extends over 500 years, is characterised by numerous significant developments: for example, the Nobel prizes which most closely relate to physics and engineering in medicine were for X-rays and radioactivity, the electrocardiogram, the scattering of radiation, the cyclotron, nuclear magnetic resonance, the transistor, radioimmunoassay and computed tomography; and a medical physicist has received the Peace Prize. The origins and development of nuclear medicine, magnetic resonance and ultrasonic imaging are representative of the whole field. Physics and engineering rank alongside other medical sciences and underpin many of their applications. In what is now the developed world, human life expectancy has increased dramatically, but the costs and risks of modern medicine have meanwhile become a huge problem. The growing divergence of rich and poor is now arguably the greatest challenge. The future cannot be predicted, but the potential of physics and engineering to improve medicine has never been greater. PMID- 11410380 TI - Evaluation of the role of ligaments, facets and disc nucleus in lower cervical spine under compression and sagittal moments using finite element method. AB - Cervical spinal instability due to ligamentous injury, degenerated disc and facetectomy is a subject of great controversy. There is no analytical investigation reported on the biomechanical response of cervical spine in these respects. Parametric study on the roles of ligaments, facets, and disc nucleus of human lower cervical spine (C4-C6) was conducted for the very first time using noninvasive finite element method.A three-dimensional (3D) finite element (FE) model of the human lower cervical spine, consisted of 11,187 nodes and 7730 elements modeling the bony vertebrae, articulating facets, intervertebral disc, and associated ligaments, was developed and validated against the published data under three load configurations: axial compression; flexion; and extension. The FE model was further modified accordingly to investigate the role of disc, facets and ligaments in preserving cervical spine motion segment stability in these load configurations. The passive FE model predicted the nonlinear force displacement response of the human cervical spine, with increasing stiffness at higher loads. It also predicted that ligaments, facets or disc nucleus are crucial to maintain the cervical spine stability, in terms of sagittal rotational movement or redistribution of load. FE method of analysis is an invaluable application that can supplement experimental research in understanding the clinical biomechanics of the human cervical spine. PMID- 11410381 TI - Improved prediction of proximal femoral fracture load using nonlinear finite element models. AB - Hip fracture, which is often due to osteoporosis or other conditions affecting bone strength, can lead to permanent disability, pneumonia, pulmonary embolism, and/or death. Great effort has been directed toward developing noninvasive methods for evaluating proximal femoral strength (fracture load), with the goal of assessing fracture risk. Previously, computed tomographic scan-based, linear finite element (FE) models were used to estimate proximal femoral fracture loads ex vivo in two load configurations, one approximating joint loading during single limb stance and the other simulating impact from a fall. Measured and computed fracture loads were correlated (stance, r=0.867; fall, r=0.949). However, precision for the stance configuration was insufficient to identify subjects with below average fracture loads reliably. The present study examined whether, for this configuration, nonlinear FE models could be used to identify these subjects. These models were found to predict fracture load within +/-2.0 kN (r=0.962). This level of precision is sufficient to identify 97.5% of femora with fracture loads 1.3 standard deviations below the mean as having below average fracture loads. Accordingly, 20% of subjects with below average fracture loads, i.e. those with the lowest fracture loads and likely to be at greatest risk of fracture, would be correctly identified with at least 97.5% reliability. This FE modeling method will be a powerful tool for studies of hip fracture. PMID- 11410382 TI - Non-linear analysis of the arterial pulsatile flow: assessment of a model allowing a non-invasive ultrasonic functional exploration. AB - Ultrasonic measurements and modelling of blood flow in large vessels allows non invasive evaluation of clinically interesting hemodynamic variables. To this aim, a non-linear mathematical model for the pulsatile arterial flow is proposed using the approximation of "local flow" theory. The model requires only measurements of instantaneous radius and centre-line blood velocity, and the knowledge of the tube distensibility to calculate blood velocity profiles, pressure gradient and wall shear stress. Evaluation of the proposed model using experimental data obtained from the literature proved that it can provide reliable results. In addition, as shown by assessing significance of various non-linear terms, results did not significantly change when a linear pressure-radius relationship was used instead of a non-linear relationship. Also, the model was found to be moderately sensitive to arterial tapering. Thus, the proposed model is suitable for a non invasive clinical arterial exploration since it only requires three measurements which can be easily and precisely obtained in vivo using ultrasonic methods: the instantaneous radius, the centre-line velocity and the mean pulse wave velocity, this last variable characterizing the tube distensibility when assuming a linear pressure-radius relationship. PMID- 11410383 TI - Effect of compliant intermediate airways on total respiratory resistance and elastance in mechanical ventilation. AB - Total respiratory resistance and elastance were estimated off-line in a sample of 60 patients undergoing mechanical ventilation by means of two regression models in order to analyse and understand a possible physiological mechanism determining differences in inspiration and expiration. The first model considered a single value for resistance and elastance over a whole breathing cycle, whereas the second model considered separate values for inspiratory and expiratory resistance and a single value for elastance. Inspiratory resistance was found to be lower than expiratory resistance, and intermediate values were obtained for resistance estimated over the whole breathing cycle. Student's t-test showed a highly significant difference between these resistance estimates, and principal components analysis demonstrated a significant increase in information when both inspiratory and expiratory resistances were used. Minor differences were found between values of elastance calculated with the two approaches. In an attempt to interpret these experimental results, a lung model incorporating the non-linear viscoelastic properties of the intermediate airways was considered. This model suggested that changes in intermediate airway volume play a significant role in breathing mechanics during artificial ventilation and indicated that inspiratory and expiratory resistance could be useful parameters for locating airway obstruction. PMID- 11410384 TI - Quantification and localisation of damage in rat muscles after controlled loading; a new approach to study the aetiology of pressure sores. AB - To obtain more insight in the aetiology of deep pressure sores, an animal model was developed to relate controlled external loading to local muscle damage. The tibialis anterior muscle (TA) and overlying skin of a rat were compressed between indentor and tibia. Loads of 10, 70 and 250kPa at skin surface were applied for 2 or 6h. During half of the 10 and 250kPa experiments interstitial fluid pressure (IFP) in the TA was measured. The TAs were excised 24h after load application. Both amount and location of damage were assessed by histological examination using a semi-automated image-processing program. In six of eleven loaded muscles damage was found. The damage was located from superficial to deep muscle tissue in a zone never exceeding the diameter of the indentor. The IFP measurements interfered with the occurrence of damage; application of 10 and 70kPa loads only caused damage when combined with IFP measurements, whereas IFP measurements increased damage at 250kPa loads. The results showed that the developed animal model can be used to provoke local damage by applying a controlled load and that the amount and location of damage can be assessed using the newly developed techniques. PMID- 11410385 TI - Development of a three-dimensional jaw-tracking system implanted in the freely moving mouse. AB - A high-resolution mandibular tracking system was designed and tested in a freely moving mouse. A sensor unit, which consisted of four small magnetic sensors, was employed to trace small magnet movements in the three-dimensional space. After the sensor's output-to-displacement transformation equations were obtained from a multiple regression analysis of pre-experimental calibration data, the magnet and the sensors were transferred to the mouse, being kept at the same configuration as determined in the calibration system. In order to measure the three dimensional jaw movements, the magnet was glued on the mandibular surface of the mouse and the sensor unit was implanted in the nasal bone. Jaw-movement trajectories were obtained as electrical signals from the sensors after being compensated by the output-to-displacement transformation equations of the sensors with a personal computer. This sensor system, applied to the freely moving mouse, could trace the jaw trajectories with an accuracy of better than 20 microm in three-dimensional space. Consequently, the typical pattern of the rhythmical jaw movements of the mouse during mastication was obtained. The mouse protruded the mandible to the most anterior position in the jaw-opening phase and retruded to it the most posterior position in the jaw-closing phase. This tracking system may also be applied to other small animals. PMID- 11410386 TI - Volterra series modelling and compensation of non-linear distortions caused by susceptibility difference artefacts related to the presence of ferromagnetic implants in magnetic resonance imaging. AB - Magnetic resonance imaging is popular in medical settings due to its unique technical characteristics. However, its full potential has been limited by imaging artefacts caused by various phenomena. Previously, a methodology was proposed to characterize and reduce artefacts caused specifically by magnetic susceptibility differences. In the present work, the Volterra series approach is suggested as an alternative method for describing non-linear distortions induced by susceptibility artefacts. A second-order Volterra series is utilized for characterizing the image non-linearities using a block-by-block processing approach. Subsequently, a corresponding second-order inverse Volterra series is applied to compensate for the quantified distortions. In addition, a technique for automatic demarcation of recoverable and non-recoverable regions in magnetic resonance images is proposed to ameliorate the developed methodology. Experimental results indicate that this approach offers improved accuracy and flexibility in reducing non-linear distortions caused by magnetic susceptibility artefacts. PMID- 11410387 TI - Clinical decision support systems for intensive care units: using artificial neural networks. AB - The paper provides an overview of applications of artificial neural networks (ANNs) to various medical problems, with a particular focus on the intensive care unit environment (ICU). Several technical approaches were tested to see whether they improve the ANN performance in estimating medical outcomes and resource utilization in adult ICUs. These experiments include: (1) use of the weight elimination cost function; (2) use of 'high' and 'low' nodes for input variables; (3) verifying the effect of the total number of input variables on the results; (4) testing the impact of the value of the constant predictor on the performance of the ANNs. The developments presented intend to help medical and nursing personnel to assess patient status, assist in making a diagnosis, and facilitate the selection of a course of therapy. PMID- 11410388 TI - Estimation of the finite center of rotation in planar movements. AB - The finite center of rotation (FCR) is often used to assess joint function. It was the purpose of this study to compare the accuracy of the procedure of Crisco et al. [4] for estimating the FCR with a procedure which uses least-squares principles. The procedures were evaluated using noisy data rotated about a known FCR. Both procedures demonstrated increasing accuracy of FCR estimation with increasing rotation angle. As the centroid of a pair of markers was moved further from the FCR, accuracy of its location decreased. Noise levels had a strong influence on FCR estimation accuracy, with the least-squares procedure being better able to cope with noise. Increasing the number of landmarks increased FCR estimation accuracy. The accuracy of the procedure of Crisco et al. [4] increased when multiple estimates of the FCR were averaged. On all of the evaluations performed, the least-squares procedure gave small improvements in the accuracy of estimating the FCR, but was not able to circumvent the inaccuracies which arise when landmarks are not appropriately positioned, numerous, or if the rotation angle is small. PMID- 11410389 TI - A single supply biopotential amplifier. AB - A biopotential amplifier for single supply operation is presented. It uses a Driven Right Leg Circuit (DRL) to drive the patient's body to a DC common mode voltage, centering biopotential signals with respect to the amplifier's input voltage range. This scheme ensures proper range operation when a single power supply is used. The circuit described is especially suited for low consumption, battery-powered applications, requiring a single battery and avoiding switching voltage inverters to achieve dual supplies. The generic circuit is described and, as an example, a biopotential amplifier with a gain of 60 dB and a DC input range of +/-200 mV was implemented using low power operational amplifiers. A Common Mode Rejection Ratio (CMRR) of 126 dB at 50 Hz was achieved without trimming. PMID- 11410390 TI - 7Li 2D CSI of human brain on a clinical scanner. AB - Lithium salts have been widely used in the treatment of mood disorders, but the mechanism of action is still not clear. In this work, a methodology for two dimensional Lithium-7 imaging on clinical systems is presented. The data were acquired using a phosphorus volume head coil that was re-tuned for the Lithium-7 frequency. A spectroscopic sequence was used to acquire the free induction decay (FID) after volume excitation using a hard pulse. The results obtained on the head of patients undergoing lithium treatment (n = 7, 0.6 mEq/l average serum level) demonstrate that images of adequate signal to noise ratio (100:1) can be obtained in acceptable imaging times (55 min) using the proposed methodology. The distribution of 7Li appears uniform in the brains of the patients studied. PMID- 11410391 TI - Subacute myocardial infarction: assessment by STIR T2-weighted MR imaging in comparison to regional function. AB - PURPOSE: Increased T2 signal intensity (SI) can be regularly observed in myocardial infarction. However, there are controversial reports about the relationship of elevated T2 SI to myocardial viability and some authors propose that high T2 SI serves as a sign of irreversible myocardial injury. This study investigates increased T2 SI compared to myocardial function in patients with reperfused subacute myocardial infarction. Preserved function was used as criterion for viability. METHODS: Ten healthy volunteers and 17 patients with myocardial infarction and patent infarct related coronary artery were examined on a 1.5 T Magnetom Vision system (Siemens). For T2-weighted MR imaging a breath hold STIR sequence with dark-blood preparation was used. Cine FLASH 2D imaging was applied to assess myocardial function. Signal-to-noise (S/N) in STIR T2 images was measured in normal and infarcted regions and subsequently identified by two independent observers. Based on a 20 segment model of the left ventricle findings were compared to regional myocardial function. RESULTS: Elevated STIR T2 SI was found in all 17 patients and observed in 27% (204/754) of segments. S/N of normal myocardium was 5.1 +/- 0.7 in volunteers and 4.9 +/- 0.8 in patients (P = NS). Infarcted myocardium presented with significantly increased S/N 12.8 +/- 1.9 (P < 0.0001). Significant transmural elevation of T2 SI was noted in 32% of segments with preserved systolic function. CONCLUSION: Increased STIR T2 SI can be observed transmurally in post-ischemic myocardial regions with preserved function. It therefore cannot be used as an exclusive marker for the non-viable region. PMID- 11410392 TI - Spectroscopic imaging of bone marrow composition in vertebral bodies. AB - The proton spectroscopic imaging technique that uses read gradient during acquisition was used for the measurement of the proton spectra in the lumbar and thoracic part of the spine of a patient with breast cancer without known skeletal metastases. The bone marrow fat/water ratios were evaluated in the same location before and after chemotherapy treatment. The results were corrected for relaxation effects. The fat/water ratios showed a significant increase as a consequence of the bone marrow degradation process due to chemotherapy. The proposed spectroscopic imaging technique offers rapid acquisition of proton spectra from large volumes of the vertebral bodies. PMID- 11410393 TI - 1H and 13C HR-MAS NMR investigations on native and enzymatically digested bovine nasal cartilage. AB - Rheumatic diseases are accompanied by a progressive destruction of the cartilage layer of the joints. Despite the frequency of the disease, degradation mechanisms are not yet understood and methods for early diagnosis are not available. Although some information on pathogenesis could be obtained from the analysis of degradation products of cartilage supernatants, the most direct information on degradation processes would come from the native cartilage as such. We have used 1H as well as 13C HR-MAS (high resolution magic angle spinning) NMR spectroscopy to obtain suitable line-widths of NMR resonances of native cartilage. 1D and 2D NMR spectra of native cartilage were compared with those of enzymatically-treated (collagenase and papain) samples. In the 1H NMR spectra of native cartilage, resonances of polysaccharides, lipids and a few amino acids of collagen were detectable, whereas the 13C NMR spectra primarily indicated the presence of chondroitin sulfate. Treatment with papain resulted only in small changes in the 1H NMR spectrum, whereas a clear diminution of all resonances was detectable in the 13C NMR spectra. On the other hand, treatment with collagenase caused the formation of peptides with an amino acid composition typical for collagen (glycine, proline, hydroxyproline and lysine). It is concluded that the HR-MAS NMR spectra of cartilage may be of significance for the investigation of cartilage degradation since they allow the fast evaluation of cartilage composition and only very small amounts of sample are required. PMID- 11410394 TI - Adaptive and self-evaluating registration method for myocardial perfusion assessment. AB - With the advent of ultra-fast MRI, it is now possible to assess non-invasively regional myocardial perfusion with multislice coverage and sub-second temporal resolution. First-pass contrast enhanced studies are acquired with ECG-triggering and breath holding. Nevertheless, some respiratory induced movements still remain. Myocardial perfusion can be assessed locally by parametric imaging methods such as Factor Analysis of Medical Image Sequences (FAMIS), provided that residual motion can be corrected. An a posteriori registration method implemented in the image domain is proposed. It is based on an adaptive registration model of the heart combining three elementary shapes (left ventricle, right ventricle and pericardium). The registration procedure is performed on a potential map derived from the distance map. To evaluate the quality of the registration procedure a superimposition score between the registration model and the contour automatically extracted in the sequence is proposed. Rigid transformation hypotheses and registration analysis provide an efficient and automatic method which allows the rejection of outlier images, such as: out of synchronisation images, out of plane acquisitions. When compared to a manual registration method, this approach reduces processing time and requires a minimal intervention from the operator. The proposed method performs registration with a subpixel accuracy. It has been successfully applied to simulated images and clinical data. It should facilitate the use of MR first-pass perfusion studies in clinical practice. PMID- 11410395 TI - Application of two-dimensional CSI for absolute quantification of phosphorus metabolites in the human liver. AB - There have recently been a number of studies dealing with the absolute quantification of concentrations of MR-visible phosphorus compounds in different tissues. The use of absolute values rather than intensity ratios may furnish additional information about metabolic changes associated with different diseases. The purpose of this study was to develop a general procedure for measuring molar metabolite concentrations and to apply it with respect to the evaluation of human liver 31P-MRS data measured using a standard slice-selective two-dimensional CSI sequence and commercial 1H/31P surface coil. The experimental determination of all surface coil-related factors influencing signal intensity was undertaken using a gradient echo imaging technique that can be adapted to commercial systems. The resulting values for healthy volunteers (N = 9) showed concentrations of PME = 2.8 +/- 1.3 mM, PDE = 9.9 +/- 2.7 mM, P(i) = 1.7+/-0.7 mM, and ATP = 3.6 +/- 0.9 mM in the human liver. The data are quite consistent with published findings. PMID- 11410396 TI - MR monitoring of tumour thermal therapy. AB - Thermal therapy of tumour including hyperthermia and thermal ablation by heat or cold delivery requires on line monitoring. Due to its temperature sensitivity, Magnetic Resonance Imaging (MRI) allows thermal mapping at the time of the treatment. The different techniques of MR temperature monitoring based on water proton resonance frequency (PRF), longitudinal relaxation time T1, diffusion coefficient and MR Spectroscopic Imaging (MRSI) are reviewed and debated. The PRF method appears the most widely used and the most efficient at high magnetic field in spite of important drawbacks. The T1 method is the easiest method of visualisation of qualitative temperature distribution and quantitative measurement seems possible in the tissue surrounding the tumour up to a temperature of 45-65 degrees C. Despite its high temperature sensitivity, application of the diffusion method in vivo is restricted due to its high motion sensitivity. The recent MRSI technique seems very promising provided acquisition times can be reduced. Results from the literature indicate that MR temperature monitoring in vivo can be achieved in vivo with a precision of about 3 degrees C in 13 s for a voxel of 16 mm3 (1.5 x 1.5 x 7 mm) in 1.5 T scanners. PMID- 11410398 TI - Functional methods for evaluation the occurrence of delayed ischemic deficit in patients with subarachnoid hemorrhage. AB - Detection of early circulatory and electrophysiologic changes due to vasospasm (VS) after subarachnoid hemorrhage (SAH) is a necessity for in-time and adequate therapeutic management. The aim of the present case report is to describe and demonstrate the results of transcranial Doppler (TCD) monitoring and brainstem auditory evoked potentials (BAEPs) examination, which indicate the development of combined circulatory insufficiency in two vascular systems. On the 14th day after SAH, TCD showed accelerated velocities in the territory of middle cerebral artery (MCA) and basilar artery (BA). BAEPs, after ipsilateral stimulation, performed on the same day verified changes of potentials on the left side with abnormal I/V amplitude ratio. These results were suggestive of brainstem dysfunction. The CT examination of the next day revealed infarction in the posterior parietal borderline zone of the left hemisphere. The patient had clinical signs of sensory aphasia, which resolved completely after one-week treatment with nimodipine. PMID- 11410399 TI - Successful treatment of aids related PML with HAART and cidofovir. AB - Progressive multifocal leukoencephalopathy (PML) is a progressive demyelinating infection of the central nervous system caused by the JC virus (JCV) in an advanced state of the acquired immunodeficiency syndrome (AIDS). Currently there are still no drugs with proven clinical efficacy against JCV, though highly active antiretroviral therapy (HAART) and the associated partial immune reconstitution have a beneficial effect on the survival time of AIDS-related PML. Various case reports have described an acceptable response under treatment with cidofovir which may be of particular benefit if HAART does not result in sufficient immune reconstitution. We report a patient with AIDS-related PML who improved under combined therapy with HAART and cidofovir. This suggests that the immediate combination of both drugs further improves the outcome of AIDS-related PML. PMID- 11410400 TI - Bromelain is an accelerator of phagocytosis, respiratory burst and Killing of Candida albicans by human granulocytes and monocytes. AB - OBJECTIVE: The aim of this study was to examine the influence of immuno modulating agents like bromelain and trypsin (e.g. Wobenzym on granulocyte and monocyte functions in healthy volunteers and patients with disorders of the humoral immuno system X-linked agammaglobulinaemia (XLA) and common variable immuno deficiency (CVID) and to find out whether the unspecific immunity could be improved by these enzymes. METHODS: In a whole-blood assay kinetics of phagocytosis, respiratory burst and killing (PBK) were measured in blood samples incubated with and without bromelain and trypsin (B/T) using Candida albicans as target organism. The time-reaction curves were analysed determining their gradient (T1) and their onset (T2) as well as the half effect time (HET). RESULTS: Phagocytes from patients with XLA showed a significantly accelerated basal phagocytosis (reduction of HET by 24% p < 0.001) compared to healthy controls. After incubation with B/T (10 microg/ml each) speed of phagocytosis was nearly doubled (phagocyte activity p < 0.0001, Candida uptake p < 0.003), T2 of respiratory burst was reduced by 65 % (p < 0.0001) and killing was accelerated by 27% (p < 0.046). However, the maximal activities of all kinetics were not altered. Incubation of phagocytes from healthy controls with B/T accelerated phagocytosis to a level comparable to that of untreated phagocytes from patients with XLA and also accelerated reactive oxygen species (ROS) production (reduction of HET by 28%, p < 0.012). In contrast to phagocytes from patients with XLA, phagocytes of patients with CVID showed a similar stimulation by B/T like healthy controls. Further experiments with the single substances showed that bromelain was the active compound. CONCLUSION: Our data suggest, that bromelain possesses immuno stimulatory properties. Phagocytes of XLA patients appear to be particularly susceptible to this stimulation. PMID- 11410401 TI - A modified system to evaluate the potency of anti-oxidative compounds in different cell types in vitro. AB - Common assays for evaluation of antioxidative capacity of different compounds are usually performed in cell-free systems. By this approach, cell-specific regulatory mechanisms upon distinct stimuli are not taken into account. Therefore, there is a need to measure anti-oxidative capacity in a cellular setting. - We now developed a valid method that provides monitoring of anti oxidative capacities of compounds in different cell types. Oxidative stress, induced by 100 microM H subset2O subset2 in human microvascular endothelial cells (HMEC-1), was quantified by the generation of oxidized, fluorescent C-DCF from C H subset2DCF-DA/AM. As DCF-production could be almost completely blocked by diethyldithiocarbamate (DEDTC), which inhibits intracellular Cu/Zn superoxide dismutase (SOD), mainly intracellular production of C-DCF was assumed. Preincubation with alpha-tocopherol resulted in a dose-dependent reduction of both spontaneous and H subset2O subset2-induced C-DCF-production (maximal inhibition by 41.6% at 75 microM). A synergistic effect was observed with co incubation with vitamin C (maximal inhibition 46.8% at 10 microM vitamin C and 50 microM alpha-tocopherol). In this way compounds with different modes of action and subcellular localization can be evaluated concomitantly in respect of their anti-oxidative capacities. As this method was established on 24- and 48-well plates in other cell lines (Caco-2, HFP-1), too, screening of a large array of antioxidative compounds in different cell lines can be performed. PMID- 11410402 TI - Isolation, purification and partial characterization of early pregnancy factor (EPF) from sera of pregnant women. AB - Early pregnancy factor (EPF) is a pregnancy protein, which is secreted into the maternal serum 12-16 hours after fertilization. It is thought to be an immunosuppressive molecule. EPF is detected in pregnant woman's serum by the rosette inhibition assay (RIA). In this study, EPF was purified from the pregnant woman's sera by using ion exchange chromatography and analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins which showed a positive result with the RIA, were found to be 35 kDa and 17 kDa molecular weights. The biological activities of these proteins were stable upon heat treatment at 56 degrees C for 30 min. Proteins isolated and purified in this study might be of great significance to the field of human reproduction with particular reference to pregnancy and recurrent abortion. PMID- 11410403 TI - Human natriuretic peptides exhibit antimicrobial activity. AB - Here we describe a novel function for members of the well-characterized human natriuretic peptide family. Human "brain-type natriuretic peptide" (hBNP-32) as well as other members of this peptide class are antimicrobially active against Gram-positive and Gram-negative bacteria and yeast in a dose-dependent manner. This activity of natriuretic peptides is comparable to that of known antimicrobial peptides such as casocidins or magainins. PMID- 11410404 TI - Quick reference guide to antiretrovirals. AB - Antiretroviral therapy continues to rapidly evolve and complex drug interactions are common. The quick reference guide is an attempt to summarize the following aspects of highly active antiretroviral treatment in concise tables: dosing of antiretroviral agents; drug-drug interactions between antiretroviral agents; antiretroviral resistance mutations and interactions of antiretroviral agents with TB-medications, oral contraceptives and methadone. Dosing and drug interaction data has been collected from manufacturers' dosing information, peer reviewed medical journals and scientific meetings. References are inserted directly in the tables referring to conferences or publication the information is derived from. In order to remain clinically relevant, regular updates to this publication are posted on the Medscape Web site at http://hiv.medscape.com/updates/quickguide - Please check regularly to ensure you are using the most recent edition. It is available as an HTML document; a PDF document; and for downloading to your Palm organizer. PMID- 11410405 TI - The early preclinical diagnosis of osteoporosis measuring the pure trabecular bone density. AB - The major skeletal determinants of fracture risk are bone mass and postmenopausal bone loss with gradual destruction of trabecular bone architecture. Therefore, the management of osteoporosis is to prevent bone loss after menopause in women below 60 years of age. We need a screening test of bone mineral density (BMD) with the ability to identify correctly individuals with accelerated bone loss; unnecessary treatment has to be avoided. But without measurable extraskeletal factors, no technique assessing BMD as single test has the reasonable high sensitivity and specificity for the diagnosis of fast bone losers. It is on principle the repeated volumetric bone measurement--within 12 months--in the pure trabecular bone, especially at distal radius of the non-dominant arm with the peripheral quantitative computed tomography (pQCT), being able to detect accurately the individuals as fast bone losers with trabecular bone loss > 8 mg/cm3 or > 3.5% per year (prevalence about 35%). Only women with very high trabecular BMD (in highest quintile) have no risk for osteoporosis. PMID- 11410406 TI - Unrelated donor marrow transplantation: an update of the experience of the Italian Bone Marrow Group (GITMO). AB - BACKGROUND AND OBJECTIVES: Unrelated donor bone marrow transplant (UD-BMT) has become an attractive alternative source of hematopoietic cells for patients lacking a matched sibling. The aim of this paper was to report on results of the 696 UD BMTs performed in 31 Italian institutions during the first 10 years of activity of the Italian Bone Marrow Donor Registry (IBMDR). EVIDENCE AND INFORMATION SOURCES: In 1989 the Italian Bone Marrow Transplant Group (GITMO) established the IBMDR to facilitate donor search and marrow procurement for patients lacking an HLA identical sibling. By end of December 1999, 260,000 HLA A, B typed volunteer donors had been cumulatively registered and 2,620 searches had been activated for Italian patients. At least one HLA-A, B, DRB1 matched donor was found for 54% of the patients and 696 UD BMTs were performed. In 50% of cases the donor was found in the IBMDR and in 50% in 15 other Registries. The average time from search activation to transplant was 6 months for disease other than CML. For CML it was 14 months. Actuarial 12-month transplant-related mortality (TRM) was 68% in patients grafted between 1979 and 1992 and 44% for patients grafted after 1993. Twenty-eight per cent of patients developed grade III or IV acute GvHD and 24% developed extensive chronic GvHD. The rate of disease free survival at three years was 57% for patients with 1st chronic phase CML, 37% for patients with 1st or 2nd CR ALL, 31% for AML or MDS patients 18 years of age and 54% for patients with inborn errors. PERSPECTIVES: We conclude that the IBMDR has benefited a substantial number of patients lacking a matched sibling and has facilitated the recruitment of UDs into the international donor pool. The long time required for the search is the major obstacle to the success of this programme. This suggests that early transplant and a decrease in TRM could further improve these encouraging results. PMID- 11410407 TI - High expression of the ILT2 (LIR-1) inhibitory receptor for major histocompatibility complex class I molecules on clonal expansions of T large granular lymphocytes in asymptomatic patients. AB - BACKGROUND AND OBJECTIVES: The lymphoproliferative disorders of large granular lymphocytes (LGLD) are divided into two groups: T-cell type and NK-cell type. These entities may be either asymptomatic or associated with autoimmune manifestations (especially cytopenias). A number of surface receptors, expressed by NK-cells and some T-lymphocyte subsets repress cytotoxicity and cytokine production upon ligation with HLA class I molecules and are clonally expressed in theses lymphoproliferative disorders. These cytotoxic lymphocytes can lyse erythroid progenitors in vitro, and the physiologic lower levels of HLA class I antigens on the erythroid lineage may contribute to this form of autoimmunity. It is conceivable that the clinical outcome of T-LGLD might be influenced by the expression of MHC class I inhibitory receptors. DESIGN AND METHODS: We analyzed the surface expression of these molecules, lectin-like heterodimers (CD94/NKG2A) or killer immunoglobulin (Ig)-like receptors (KIR) and another Ig-like inhibitory receptor, termed ILT2 or LIR-1 in CD8+ cells from 12 cases of ab T-LGLD using specific monoclonal antibodies. RESULTS: None of the LGLD cases had anemia and 11 of 12 patients remain asymptomatic. KIR and CD94/NKG2A expression was detected on CD8+ populations only in some cases of T-LGLD. By contrast, our observations revealed that ILT2 expression was markedly higher in CD8+ cells from LGLD patients than from healthy donors. INTERPRETATION AND CONCLUSIONS: Expression of the ILT2 inhibitory receptor for HLA class I molecules on LGLD cells might indeed contribute to preventing their autoreactivity. Further studies are required to evaluate the expression/function of the ILT2 receptor in patients who eventually become symptomatic. The development of cytopenias in LGLD patients must involve other self-reactive activating receptors. Analysis of the expression and function of triggering NKR in LGLD needs to be carefully addressed. PMID- 11410408 TI - Polycythemia vera: analysis of DNA from blood granulocytes using comparative genomic hybridization. AB - BACKGROUND AND OBJECTIVES: The diagnosis of polycythemia vera (PV) is supported by the finding of an abnormal karyotype in patients with erythrocytosis. However, most PV patients have normal marrow cytogenetics at presentation and there is reluctance to use this test routinely. Comparative genomic hybridization (CGH) is a cytogenetic screening technique that analyzes interphase cells. This approach offers practical advantages over conventional cytogenetics and interphase fluorescence in-situ hybridization (IFISH). We have therefore evaluated the diagnostic utility of CGH applied to blood granulocytes in PV. DESIGN AND METHODS: Blood granulocytes from 17 PV patients were analyzed using CGH and the results compared with those from previous conventional cytogenetics and IFISH studies. RESULTS: Three patients had abnormal CGH profiles. One case had gain of 9p. This patient had normal IFISH results using a centromere-9 probe. The second case had complete gain of chromosomes 8 and 9 and the third had complete gain of chromosome 9, all confirmed by IFISH: Cytogenetics had not been performed in two of these cases and had failed in the third. Three cases with 20q deletion according to cytogenetics and/or IFISH, were normal by CGH. The remaining subjects were normal by all methods. INTERPRETATION AND CONCLUSIONS: CGH analysis of blood granulocytes can detect the chromosome gains commonly observed in PV. However, CGH cannot be relied on to detect 20q deletions, which are the most frequent cytogenetic abnormality in PV. Thus, CGH has a role in the diagnosis and follow-up of PV patients, but must be used in conjunction with other methods. PMID- 11410409 TI - The prognostic value of Bcl-XL gene expression for remission induction is influenced by cytogenetics in adult acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: There is growing evidence that altered expression of genes belonging to the BcL-2 family of apoptosis regulators might influence chemotherapy-induced apoptosis in malignant cells and therefore could confer multidrug resistance. So far expression studies of apoptosis-regulating genes on acute myeloid leukemia (AML) have mainly focused on Bcl-2 itself and most of them have not included other factors involved in drug resistance or apoptosis as parameters determining response to chemotherapy, disease progression and survival. DESIGN AND METHODS: We therefore examined Bcl-2, Bcl-XL and Bax gene expression in 235 adult patients with de novo or secondary myeloid leukemia. The expression levels were correlated with established prognostic factors such as age, cytogentic aberrations, mdr1 gene expression and clinical outcome in a multivariate analysis. RESULTS: Bcl-2 and Bcl-XL positive patients had a much lower white blood cell count than negative patients (p<0.001 and p=0.003, respectively). Bcl-2 expression correlated with FAB subtype M0 (p=0.03), Bax with M5b (p=0.02) and Bcl-XL with M6 (p=0.005). Mdr1 expression was more frequently seen in Bcl-2 and Bcl-XL positive patients (p=0.03 and p=0.02, respectively). Remarkably Bax was significantly less frequently expressed in de novo AML patients with high risk cytogenetics (p=0.007). No difference in expression was recognized for Bcl-2 or Bcl-XL when statistical analyses were done for cytogenetic risk groups. However, in the multivariate analysis regarding the group of de novo AML patients < or =60 years with intermediate risk cytogenetics, Bcl-XL expression was found to be an independent negative prognostic factor for response to induction therapy (p=0.04). In contrast, no prognostic impact of Bcl XL expression on treatment response was seen within the group of patients with high risk cytogenetic findings. Neither Bcl-2 nor Bax nor Bcl-XL expression had a significant influence on overall or disease-free survival. INTERPRETATION AND CONCLUSIONS: These data indicate that the prognostic value of Bcl-XL gene expression for treatment response in AML patients < or =60 years is dependent on cytogenetics. PMID- 11410410 TI - Treatment of childhood acute lymphoblastic leukemia. Long-term results of the AIEOP-ALL 87 study. AB - BACKGROUND AND OBJECTIVES: In March 1987 AIEOP started the AIEOP-ALL-87 study, based on the previous AIEOP-ALL-82. The aim of this new study was to evaluate, for all risk groups: a) the efficacy of treatment intensification achieved by adding a fourth drug (daunomycin) in the induction phase and a 3-drug reinduction phase for all risk groups; b) the impact of the addition of three doses of intrathecal methotrexate during cranial radiotherapy and extended exposure to weekly high-dose L-aspariginase during late intensification in high risk patients. We report the long-term results of the AIEOP ALL-87 study. DESIGN AND METHODS: From 1987 to 1991, a total of 632 eligible and evaluable children (age 1 to < or =16 years) with non-B-cell acute lymphoblastic leukemia (ALL), were enrolled and stratified as follows: standard risk (SR, 79 patients, 12.5%) had WBC <10,000/mm3, age > or = 3 and <7 years, and FAB L1 morphology. The high risk (HR, 175 patients, 27.7%) group included patients with WBC > or =50,000/mm3 or FAB L3 morphology or T immunophenotype or acute undifferentiated leukemia (AUL) or leukemia-lymphoma syndrome. All the remaining patients formed the intermediate risk group (IR, 378 patients, 59.8%). All patients received a 4-drug induction therapy; intermediate-dose methotrexate was given to HR patients; cranial radiotherapy was given to IR and HR patients, while SR patients received extended intrathecal methotrexate; all patients received a 3-drug reinduction phase; high dose L-asparaginase (HD-L-ASP; E.Coli, Bayer) was given to HR patients; continuation therapy with 6-mercaptopurine, i.m. methotrexate, and monthly vincristine and prednisone pulses was given to all patients. Treatment duration was 2 years. RESULTS: Six hundred and nineteen patients (97.9%) achieved complete remission. The remission rate was 98.7% in the SR group, 98.1% in the IR group, and 97.1% in the HR group. The overall 10-year survival and event-free survival (EFS) rates (SE) are 74.7% (1.8) and 62.8% (2.0) respectively; EFS rates by risk group are 67.5% (5.5) in SR, 62.8% (2.6) in IR, and 61.9% (3.8) for HR. The 10 year EFS for all eligible patients was 63.9% (1.9). INTERPRETATION AND CONCLUSIONS: When compared to the results of the AIEOP-ALL-82 study, treatment intensification in the ALL-87 study has improved long-term survival and EFS from 66.4% and 53.6% to 74.7% and 62.8%, respectively. Failures were mostly due to marrow or extramedullary relapses suggesting that further treatment intensification, as being used in current therapeutic strategies, is appropriate, although patients relapsing after less intensive treatment may have better chances of rescue. These results, although obtained in a relatively large proportion of patients, in which infants were not included, indicate that the addition of high-dose L-asparaginase to a relatively non-intensive treatment may be of major benefit for HR patients and that the addition of intrathecal methotrexate during CRT, may improve the central nervous system-disease control with a marked reduction of nervous system relapses. PMID- 11410411 TI - In vitro induction of apoptosis of neoplastic cells in low-grade non-Hodgkin's lymphomas using combinations of established cytotoxic drugs with bendamustine. AB - BACKGROUND AND OBJECTIVES: Regulation of apoptotic cell death is being increasingly recognized as a mechanisms by which cytostatic agents mediate tumor cell death. Preliminary clinical studies with bendamustine, an alkylating agent with a purine nucleus, provide strong evidence that this drug is a highly effective cytostatic in low grade lymphomas. Therefore, we investigated the in vitro activity of bendamustine in combination with other established cytotoxic drugs. DESIGN AND METHODS: 2 lines (DOHH-2, WSU-NHL) and mononuclear cells (MNC) from patients with leukemic low-grade B-non-Hodgkin's lymphoma (NHL) (n=10), T NHL (n=7) and chronic lymphocytic leukemia (CLL) (n=12). Apoptosis (7-AAD), depolarization of mitochondrial membrane potential (MMP, JC-1), caspase-3 activity (FIENA) and cell proliferation (XTT/WST-1) were determined. Several incubation times and drug dosages (for IC(30/50/75/90)) were studied. Synergistic, additive or antagonistic effects were calculated by a median plot effect and the combination index (CI) method. RESULTS: In general, combinations of bendamustine with mitoxantrone or doxorubicin resulted in antagonistic effects in the tested cell lines and the MNC from the patients. CI-calculation failed in these cases since there was not a sufficient dose response. On the other hand, the combination of bendamustine with 2-CdA showed synergistic in vitro activity on the tested cell lines, neoplastic lymphocytes from patients with peripheral T cell lymphomas and partially on MNC from patients with CLL and B-NHL. The antagonism of the combination of bendamustine and anthracyclines appeared to be due to inhibition of depolarization of mitochondrial-membrane potential and caspase-3-activity during apoptosis of the studied cell lines. INTERPRETATION AND CONCLUSIONS: In conclusion, our results suggest that schedules using combinations of bendamustine and anthracyclines should not be recommended for the treatment of low-grade NHL, whereas bendamustine combined with 2-CdA could be considered for the development of future treatment strategies. PMID- 11410412 TI - Identification of factors associated with poor peripheral blood progenitor cell mobilization in Hodgkin's disease. AB - BACKGROUND AND OBJECTIVES: Although the use of drugs which damage stem cells is common in patients with Hodgkin's disease (HD), factors affecting peripheral blood progenitor cell (PBPC) mobilization have not been clearly established in this group of patients. The aim of this study was to identify factors associated with poor PBPC mobilization in patients with HD. DESIGN AND METHODS: In order to address this issue we have evaluated in 54 patients with HD mobilized with G-CSF alone the following factors: sex, age, histologic subtype, B symptoms at diagnosis, status of remission, previous chemotherapy and radiotherapy, interval from diagnosis and last chemotherapy cycle to harvest, and dose of G-CSF. Univariate analysis was performed using Student's t-test, Pearson's correlation and Spearman's correlation. A stepwise regression model was used to determine which of the variables was the most predictive of PBPC mobilization. RESULTS: In univariate analysis poorer PBPC mobilization was observed in patients who had previously received at least two courses of mini-BEAM (p=0.006), a high number of different chemotherapy regimens (p=0.002), a chemotherapy score >30 (p=0.02) and more than 9 months of alkylating agents (p=0.07). We did not find radiotherapy to be a significant factor affecting progenitor cell yield (p=0.59). In the stepwise regression model, only the previous administration of two or more mini-BEAM cycles predicted a poor PBPC yield (p=0.006). INTERPRETATION AND CONCLUSIONS: Previous chemotherapy, principally exposure to a mini-BEAM regimen, seems to be the principal factor affecting collection of PBPC in patients with HD mobilized with G-CSF alone. Since mini-BEAM is an effective salvage regimen in relapsed or refractory HD, collection of PBPC should be planned when there has been no or only minimal exposure to a mini-BEAM regimen. PMID- 11410413 TI - Familial hemophagocytic lymphohistiocytosis: how late can the onset be? AB - BACKGROUND AND OBJECTIVES: Most patients with familial hemophagocytic lymphohistiocytosis (HLH) develop the disease within the first two years of age. In a minority of cases a later occurrence has been reported, with an upper age limit of eight years. A significant concordance of the age at onset within each family has also been observed. RESULTS: We report four cases of families with HLH diagnosed at an unusually late age, comprised between between 9 and 17 years; in each of these families another child developed the disease in infancy. The natural killer activity of the patients was depleted; nevertheless, we had indirect evidence that, in at least two families, mutations of the perforin gene were not causing the disease. INTERPRETATION AND CONCLUSIONS: Such a late onset is very unusual and suggests that there is a subgroup of families with HLH in which the disease may present early or late in different members. Thus in some families with HLH the siblings might remain at risk of developing the disease for several years. Their actual risk cannot be defined until the genetic mutation is identified in each family and assessed in each member. PMID- 11410414 TI - Long-term follow-up of idiopathic thrombocytopenic purpura in 310 patients. AB - BACKGROUND AND OBJECTIVES: Idiopathic thrombocytopenic purpura (ITP) induces thrombocytopenia by means of an autoimmune mechanism. Despite the available therapies a subset of patients develop chronic refractory severe thrombocytopenia (i.e. a platelet count consistently lower than 20 to 30x10(9)/L), and life threatening bleeding can occasionally occur. It has been suggested that the risk of major bleeding is higher in elderly patients and in patients with bleeding at diagnosis. However, since clear data on the influence of clinical and/or laboratory parameters on outcome are lacking, some patients may be receiving unnecessary treatment. DESIGN AND METHODS: We made a retrospective analysis of a series of 310 patients with chronic ITP (108 males and 202 females), with a median age at diagnosis of 40 years (range 8-87 years). The median follow-up time was 121 months, (range 7-434 months). Therapy was most often started in the presence of hemorrhagic complications and/or a platelet count <30x10(9)/L either at diagnosis or during follow-up. RESULTS: Our findings confirmed that patients who were symptomatic at diagnosis were more likely to have bleeding during their follow-up. Moreover, all the patients who suffered major bleeding during their follow-up had median platelet counts of 10x10(9)/L (range 1-20) at that time. Only one patient, aged 43 years, died of hemorrhage following prolonged severe thrombocytopenia. Age >60 years was not associated with any significant differences in incidence of bleeding at diagnosis or during follow-up. INTERPRETATION AND CONCLUSIONS: We conclude that prospective studies are required to evaluate whether it may be reasonable to treat only symptomatic patients, independently of age. PMID- 11410415 TI - Identification of polymorphisms in the 5'-untranslated region of the TAFI gene: relationship with plasma TAFI levels and risk of venous thrombosis. AB - BACKGROUND AND OBJECTIVES: Thrombin activatable fibrinolysis inhibitor (TAFI) plays an important role in hemostasis, functioning as a potent fibrinolysis inhibitor. TAFI gene variations may contribute to plasma TAFI levels and thrombotic risk. DESIGN AND METHODS: We sequenced a 2083-bp region of the 5' regulatory region of the TAFI gene in 127 healthy subjects searching for variations, and correlated identified polymorphisms with plasma TAFI levels. TAFI polymorphisms were examined as risk factors for venous thrombosis by determining their prevalence in 388 patients with deep venous thrombosis (DVT) and in 388 controls. RESULTS: Seven novel polymorphisms were identified: -152 A/G, -438 A/G, -530 C/T, -1053 T/C, -1102 T/G, -1690 G/A, and -1925 T/C. -152 A/G, -530 C/T and 1925 T/C were found to be in strong linkage disequilibrium, as were the -438 A/G, -1053 T/C, -1102 T/G and -1690 G/A. Plasma TAFI levels were higher in -438GG/ 1053CC/-1102GG/-1690AA homozygotes than in -438AG/-1053TC/-1102TG/-1690GA heterozygotes, and -438AA/-1053TT/-1102TT/-1690GG homozygotes had the lowest TAFI levels (p=0.0003). TAFI concentrations in -152AA/-530CC/-1925TT homozygotes were somewhat higher but not significantly different from levels observed for -152AG/ 530CT/-1925TC heterozygotes. Taken in combination, -438AG/-1053TC/-1102TG/-1690GA and -438AA/-1053TT/-1102TT/-1690GG yielded an OR for DVT of 0.8 (95%CI: 0.6-1). In subjects aged <35 years the OR was 0.7 (95%CI: 0.5-1.1). The OR for -152AG/ 530CT/-1925TC was 1 (95%CI: 0.5-2.2) in the whole group of patients and controls, whereas in subjects aged <35 years the OR was 0.1 (95%CI: 0.02-0.9). INTERPRETATION AND CONCLUSIONS: Polymorphisms in the TAFI promoter determine plasma antigen levels and may influence the risk of venous thrombophilia. PMID- 11410416 TI - Unmanipulated bone marrow transplantation from one-HLA locus mismatched siblings carries high transplant-related mortality in Chinese patients. AB - BACKGROUND AND OBJECTIVES: We compared the outcome of bone marrow transplantation (BMT) from HLA-identical siblings (MSD) and one HLA-locus mismatched siblings (PMSD) in Chinese patients with hematologic malignancies in terms of transplant related mortality (TRM) and disease relapse to see whether PMSD can feasibly increase the availability of donors in our population. DESIGN AND METHODS: Medical records of patients who had received a BMT from sibling donors in the Queen Mary Hospital, Hong Kong, from March 1990 to February 2000 were reviewed (MSD 326, PMSD 20). Patients and their donors were matched for HLA-A, -B and DRB1 loci using standard serologic methods as well as polymerase chain reaction sequence specific primers. All patients received standard anti-microbials and graft -versus host disease (GVHD) prophylaxis including cyclosporin A and a short course of methotrexate. RESULTS: A total of 346 BMT patients were analyzed of whom 326 and 20 patients had received transplants from matched and one locus mismatched siblings, respectively. Patients receiving BMT from PMSD had a significantly higher TRM than those receiving their BMT from MSD (p=0.0016). Six patients received BMT from HLA-DR PMSD: one died 2 months post-BMT as a result of post-transplantation-related lymphoproliferative disease. Fourteen patients received BMT from HLA-A or -B PMSD: 11 of these patients died after a median of 5.6 months (range 0.6-13.7 months) due to severe GVHD (n=5), graft failure (n=2), bleeding (n=1), leukemic relapse (n=2) and thrombotic thrombocytopenic purpura (n=1). Two out of the three survivors had primary graft failure: one of these two required infusion of back-up marrow and the other had autologous regeneration. Patients in the PMSD group were at greater risk of developing severe GVHD than their MSD-recipient counterparts (p<0.001). There was no significant difference in the probability of disease relapse between patients who received BMT from MSD or PMSD. INTERPRETATION AND CONCLUSIONS: BMT from PMSD (especially those with mismatches at HLA class I loci) carried a higher risk of TRM and morbidity than BMT from MSD in our population. PMID- 11410417 TI - Neoadjuvant high dose chemotherapy plus peripheral blood progenitor cells in inflammatory breast cancer: a multicenter phase II pilot study. AB - BACKGROUND AND OBJECTIVES: With the introduction of combined modality therapy, approximately 30% of patients with inflammatory breast cancer (IBC) are alive and free of disease at 5 years, but the lack of control of systemic disease continues to be the main reason for treatment failure. The importance of the response to primary chemotherapy and, in particular, complete tumor regression after primary chemotherapy have previously been described to be among the most reliable prognostic factors along with the dose intensity of doxorubicin. DESIGN AND METHODS: To evaluate pathologic response rate and toxicity of neoadjuvant high dose chemotherapy (HDCT) with autologous peripheral blood progenitor cell (PBPC) support in patients affected by IBC, 21 patients were enrolled in a study in which it was planned that they would receive 4 courses of epirubicin 150 mg/m(2) plus granulocyte colony-stimulating factor (G-CSF) as induction and mobilizing chemotherapy. Patients with non-progressive disease were intended to receive 2 consecutive courses of a combination of high doses of mitoxantrone 40 mg/m(2) , thiotepa 500 mg/m(2) and cyclophosphamide 200 mg/kg as a conditioning regimen. RESULTS: PBPC collection was successful in 20/21 patients. Twelve patients received a single course of HDCT, whereas 7/20 patients underwent a double procedure. At a median follow up of 48 months, 20/21 patients were evaluable for toxicity and 19/21 for response. At surgery 4/19 patients (21%) had no evidence of viable tumor cells in the breast and in axillary nodes, while 4 (21%) and 11 patients (58%) had microscopic and macroscopic disease, respectively. Eight patients have relapsed (35%) so far at a median of 16 months (9-54) from diagnosis. Eleven patients remain alive without evidence of disease. Five out of 20 patients experienced severe cardiotoxicity with congestive heart failure (CHF) which was responsible for the only treatment-related death. INTERPRETATION AND CONCLUSIONS: This neoadjuvant HDCT regimen seems to be very effective in terms of objective responses, but we observed a high rate of cardiotoxicity and only a few patients were able to receive the two planned courses of high dose chemotherapy. PMID- 11410418 TI - Effect of cold-storage in the accumulation of bioreactive substances in platelet concentrates treated with second messenger effects. AB - BACKGROUND AND OBJECTIVES: The mandatory 5-day of shelf-life platelet concentrates (PCs) creates outdating and inventory control problems in blood banking. Moreover, storage of PCs at 22-24 degrees C has been associated with a time-dependent accumulation of pyrogenic cytokines, potentially harmful for recipients. Previous studies have shown that supplementation of PCs with ThromboSol, a mixture of second-messengers effectors, might allow storage of functionally active platelets at refrigerated temperature to be extended. This study further investigates this storage approach by comparing the accumulation of bioactive compounds in standard and refrigerated PCs. DESIGN AND METHODS: The PCs were supplemented with ThromboSol or a control solution and stored in parallel at 24 degrees C with continuous agitation or undisturbed at 4 degrees C. Samples were removed on days 1, 5, 9 of storage, and assayed for their content of interleukin (IL)-6, IL-8, tumour necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta1, and anaphylatoxins C3a and C4a. RESULTS: Throughout storage, refrigerated PCs, both ThromboSol-treated and untreated units, displayed a slightly lower level of IL-6 and significantly lower concentration of IL-8 than conventionally stored PCs. ThromboSol slightly reduced the level of these cytokines in PCs. Throughout storage at 22 degrees C, an accumulation of anaphylatoxins C3a and C4a was seen both in both control and ThromboSol-treated PCs. This accumulation was significantly reduced in control PCs stored at 4 degrees C, but not in refrigerated PCs supplemented with ThromboSol. Cold storage, with or without ThromboSol, had a minor effect on the accumulation of TGF-beta1 in PCs. INTERPRETATION AND CONCLUSIONS: Our data confirm that release of bioactive compounds during in vitro storage of PCs is a temperature-sensitive process. The ThromboSol-refrigeration system could be a useful alternative for extending storage of PCs, without increasing the accumulation of cytokines (IL-6, IL-8), known to be involved in febrile reactions in recipients. Nevertheless, this storage system has no benefit on the level of other bioactive compounds (TGF beta1, anaphylatoxins C3a and C4a) in PCs. PMID- 11410419 TI - Alpha128 Arg-->Ser (CGT-->AGT) spectrin mutation associated with severe neonatal elliptopoikilocytosis in Spain. PMID- 11410420 TI - Interaction between (--SEA) alpha-thalassemia deletion and uncommon non deletional alpha-globin gene mutations in Chinese patients. PMID- 11410421 TI - Alpha2(CD31 AGG-->AAG, Arg-->Lys) causing non-deletional alpha-thalassemia in a Chinese family with HbH disease. PMID- 11410422 TI - Is polycythemia vera a serious disease in young adults? PMID- 11410423 TI - Persisting molecular remission ten years after donor lymphocyte infusion for hematologic relapse in chronic myeloid leukemia. PMID- 11410424 TI - TEL rearrangements in acute lymphoblastic leukemia: association with p16 deletions in relapsed cases. PMID- 11410425 TI - Intracellular interleukin-2 expression by T-cell subsets in B-cell chronic lymphocytic leukemia. PMID- 11410426 TI - Successful treatment of acquired hypoprothrombinemia without associated lupus anticoagulant using intravenous immunoglobulin. PMID- 11410427 TI - An affordable Fc-receptor blockade method to treat patients with chronic refractory autoimmune thrombocytopenic purpura. PMID- 11410428 TI - Effects of clarithromycin on oral mucositis in bone marrow transplant recipients. PMID- 11410429 TI - Acute transverse myelitis and autoimmune pancytopenia after unrelated hematopoietic cell transplantation. PMID- 11410430 TI - The irreplaceable image: Serous degeneration of the bone marrow. PMID- 11410431 TI - The irreplaceable image: Black tea delays recovery from iron-deficiency anemia. PMID- 11410432 TI - The irreplaceable image: Non-Hodgkin's lymphoma in a totally implantable subcutaneous venous access port system. PMID- 11410433 TI - Choosing a treatment strategy for atrial fibrillation. PMID- 11410434 TI - Is wine the healthiest form of alcohol? PMID- 11410435 TI - Iron and heart-disease risk. PMID- 11410436 TI - Ask the Doctor. I know that there have been a lot of studies showing that beta blockers help patients with congestive heart failure live longer. I have heart failure, and last year my doctor prescribed a beta blocker. In a few days, I became so short of breath that he had me stop taking it. I am upset that I can't use this important therapy. Is there any way I might be able to do so? PMID- 11410437 TI - Ask the Doctor: I was very upset to read about the high rates of abnormal brain function that follow cardiac surgery. I've had two angioplasties, and my doctors say that I may need to have bypass surgery in the future. If I undergo this operation, is there anything I can do to reduce my chances of having a stroke or a memory problem afterward? PMID- 11410438 TI - Medical care. Home tests. PMID- 11410439 TI - Mad cow disease and chondroitin sulfate. PMID- 11410440 TI - The doctor-patient relationship. Doctors from Venus and Mars: how they differ. PMID- 11410441 TI - Coronary artery disease. Do stents keep arteries open? PMID- 11410442 TI - Puzzling through a case. The snoring accountant. PMID- 11410444 TI - By the way, doctor...I've been trying to lose weight on my own for years. I'm interested in trying something over the Internet, partly because it seems more private than other weight-loss programs. Any chance an Internet program could work? PMID- 11410443 TI - Aging. A conversion about frailty. PMID- 11410445 TI - Bipolar disorder -- Part II. PMID- 11410447 TI - New treatments for cocaine addiction. PMID- 11410446 TI - Steroids: youth at risk. PMID- 11410448 TI - Attachment and the therapeutic relationship. PMID- 11410449 TI - Exercise and the prostate: can you run away from trouble? PMID- 11410450 TI - Peripheral artery disease. Part II: diagnosis and treatment. PMID- 11410451 TI - Medical memo. Climbing the stairway to health. PMID- 11410453 TI - Breast cancer and fruits and vegetables. PMID- 11410452 TI - On call. At 44 years of age, I am 6 feet tall and weigh 210 pounds. I feel fine but I get very sleepy during the day. Am I overweight - and is that why I'm so sleepy? PMID- 11410454 TI - Celiac disease and how to live with it. PMID- 11410455 TI - A conversation about cancer pain. PMID- 11410456 TI - Herbal treatment for PMS? PMID- 11410457 TI - By the way, doctor. I recently heard that I can take Fosamax once a week for osteoporosis, rather than every day. Is it really effective when taken this way? Is there a downside? PMID- 11410458 TI - Strong bones: a balancing act. PMID- 11410459 TI - Developmental morphology of ovules and seeds of Nymphaeales. AB - Ovule and seed development in six species of Nymphaeales was examined. In the Cabombaceae the two species studied resemble some extant basal angiosperms by having a hood-shaped outer integument. A micropyle-hilum complex results when the outer integument and derived testa are lacking between the micropyle and the funiculus, thus the hood-shaped appearance. In the Nymphaeaceae the outer integument is annular at an early stage and then cup-shaped though it is semiannular at initiation in Nupar japonicum and Nymphaea alba. The micropyle and hilum are separated by an intervening testa. Developmental data on the formation of the outer integument, from semiannular to hood-shaped vs. from annular to cup shaped, are useful for inferring the morphology of the outer integument from the relative position of the micropyle to the hilum in seed fossils. The oldest (early Cretaceous) probable nymphaealean seeds had the micropyle-hilum complex, suggesting that the hood-shaped outer integument may be primitive in the Nymphaeales. This needs to be tested by examination of this feature in other groups of basal angiosperms. PMID- 11410460 TI - Effect of aquatic weeds on methane emission from submerged paddy soil. AB - Paddy fields are one of the dominant anthropogenic sources of methane emission to the atmosphere, and the main passageway of methane from paddy soil is through the rice plant. However, the effect of aquatic weeds on methane emission from rice paddies has not been properly evaluated yet. Methane emission from weeded pots and unweeded ones with anaerobic paddy soil was measured throughout the period of rice growth. More than double the amount of methane was emitted from weeded pots compared with unweeded ones. Peroxidase activity of rice root was not different between weeded and unweeded pots. However, methanogenic bacteria populations were higher in weeded pots than in unweeded ones, while methane oxidation activity, measured by the propylene oxidation technique, was higher in unweeded pots than in weeded ones. Methane oxidation activity of roots from three typical aquatic weeds in paddy fields, Lipocarpha sp., Rotala indica, and Ludwigia epilobioides, was higher than that of rice plants, while lower stems of these aquatic plants showed similar or lower activity compared with the same areas of rice plants. These results indicate that the role of aquatic weeds in paddy soil in methane emission should not be overlooked in evaluating mitigation options for reducing methane emission from paddy fields. PMID- 11410461 TI - Preformation, architectural complexity, and developmental flexibility in Acomastylis rossii (Rosaceae). AB - The duration of preformation and the seasonal pattern of development were studied in the architecturally complex alpine perennial Acomastylis rossii. Each leaf and inflorescence requires 3 yr to progress from initiation through structural and functional maturity to senescence. As a consequence, three cohorts of preformed organs, initiated in successive years, are borne simultaneously by each individual plant. The oldest cohort matures immediately following snowmelt, after which no additional leaves are matured until the following spring. A second cohort remains below ground in the apical bud and continues development, while a third cohort is initiated. Initiation and development of primordia proceed below ground throughout the summer and continue for at least 2.5 mo after aboveground structures have senesced. Acomastylis rossii maintains numerous dormant vegetative buds containing preformed leaf primordia in the axils of senesced leaves. Developmental preformation has been widely reported in arctic and alpine tundra environments and has been theorized to severely constrain rapid responses to environmental variation. The presence of many such preformed structures may mitigate some of the constraint on plant response to environmental variation imposed by the long developmental trajectories of leaves and inflorescences in apical buds. PMID- 11410462 TI - Ontogenetic derivation and cell differentiation in photosynthetic tissues of C3 and C4 Cyperaceae. AB - Four variants of Kranz anatomy occur in the Cyperaceae. Three of these anatomical types (fimbristyloid, chlorocyperoid, and eleocharoid) are unique among taxa with C(4) photosynthesis in that the photosynthetic carbon reduction tissue (PCR, functional equivalent of bundle sheath) is located within the vascular strand and is separated from the primary carbon assimilation tissue (PCA, positional equivalent of mesophyll) by the mestome sheath layer. In the fourth anatomical type, rhynchosporoid, PCR tissue is located in the position of the mestome sheath. In this study, we compared two aspects of development of PCR and PCA tissues in representatives of the C(3) and C(4) types: (1) ontogenetic derivation and (2) cellular differentiation. Analysis of the planes of cell division associated with procambial strand formation indicated that PCR tissue is always derived from the procambium, while PCA tissue is derived from the ground meristem. These cell lineages remain distinct after the initial organization of vascular strands. Analysis of cell differentiation using accumulation of cell type-specific photosynthetic enzymes as markers of differentiation indicated that, with one exception, a low level of non-cell-specific enzyme accumulation preceded abundant and cell-specific accumulation of photosynthetic enzymes at the distal end of the leaf elongation zone. Enzyme accumulation coincided spatially (and temporally) with structural aspects of cell differentiation. Previous cladistic analyses have indicated that these anatomical types represent separate evolutionary origins of the C(4) pathway, and the differences in developmental pathways observed here reflect these independent origins from C(3) ancestors. PMID- 11410463 TI - Spatial genetic structure of allozyme polymorphisms within populations of Pinus Strobus (Pinaceae). AB - The population structure of genetic variation for four allozyme loci was investigated for two populations (one old growth, OG, and the other logged, SS) of eastern white pine (Pinus strobus). Both seedlings and reproductive adults were studied for both study populations. Spatial autocorrelation statistics were used to examine the distribution of allozyme polymorphisms. The spatial genetic structure in adults of population OG indicated that individual genotypes were distributed in a structured, isolation-by-distance manner, consistent with observed levels of pollen and seed dispersal. In contrast, adult genotypes in population SS were nearly randomly distributed, probably as a result of logging. Nonetheless, spatial structuring of genotypes of seedlings occurred at both sites, indicating the power of limited seed flow, as well as temporal Wahlund effects, to create structure. None of four loci in both seedling populations showed a significant departure from Hardy-Weinberg proportions, whereas one and two significant deviations were found for loci in the two respective adult populations. These departures may be attributed to episodic reproductive events. PMID- 11410464 TI - Pollen limitation of reproductive success in two sympatric alpine willows (Salicaceae) with contrasting pollination strategies. AB - We compared the extent of pollen limitation on female reproductive success of Salix lanata L., an entirely insect-pollinated willow, and S. lapponum L., which is 50 : 50% insect : wind pollinated (ambophilous). Supplemental hand-pollination significantly increased seed number per fruit by nearly 50% in the insect pollinated willow, but had no significant impact on seed number in the dually pollinated species. Fruit set was not affected by the treatment in either of the species. These results demonstrate that pollen limitation on reproductive success is most pronounced in the species that depends entirely on insects for pollination. In general, pollinator visitation was highest to S. lapponum, but bumble bees were only observed on S. lanata, suggesting that the quantity and quality of pollinator visitation differed between the species. Our results empirically support the hypothesis that a dual pollination strategy is most effective in alpine environments with low and infrequent pollinator activity and high wind speeds. PMID- 11410465 TI - Neither vegetative nor reproductive advantages account for high frequency of male steriles in southern Spanish gynodioecious Daphne laureola (Thymelaeaceae). AB - In gynodioecious species, male-steriles (termed "females" hereafter) usually exhibit some reproductive advantage over hermaphrodites that allow them to compensate for the loss of male reproductive function. This compensation can result from higher fecundity, vegetative outperformance, and/or lower inbreeding depression. In this study we compared vegetative and reproductive parameters of female and hermaphrodite Daphne laureola individuals in two southeastern Spanish populations and estimated the magnitude of inbreeding depression up to the seedling emergence stage by conducting controlled pollinations and experimental sowings of seed progenies in the field. Reproductive shrubs of both sexes did not differ significantly in size, leaf production, leaf size, leaf growth, nutrient allocation to leaves, and production of flowers and fruits. Seed set and seed size of cross- and self-pollinated flowers of hermaphrodites and cross-pollinated females were also similar. Seedling emergence rates of self- and cross-pollinated seeds from hermaphrodites under natural field conditions were similar, suggesting a minor role of inbreeding depression up to that reproductive stage. Seeds from females produced more seedlings than selfed seeds from hermaphrodites. In 21 populations surveyed in the study region over a broad geographical and elevational gradient, the proportion of females ranged between 20.6 and 56.1% and was inversely related to elevation. The establishment and maintenance of females in southeastern Spanish populations of D. laureola seem to be mainly mediated by ecological factors. PMID- 11410466 TI - Geographical variation in autonomous self-pollination levels unrelated to pollinator service in Helleborus foetidus (Ranunculaceae). AB - Autonomous self-pollination may be considered as a mechanism enhancing plant reproductive success when plant access to pollen sources may limit seed production. We have studied the relationship between geographical patterns of variation in pollinator service to Helleborus foetidus and self-pollination ability in three widely spaced regions in the Iberian Peninsula. As could be expected from its early flowering period, pollinator visitation rates to both plants and flowers of H. foetidus were very low at all sites. Pollinator composition remained consistent among regions, but there was significant variation among regions in pollinator service. Despite the low visitation rates, fruit set did not appear to be pollen limited in any of the study areas, which may be explained by the long duration of flowers (up to 20 d). When pollinators were excluded experimentally, fruit set decreased significantly, but substantial levels of self-pollination occurred at all regions. Autonomous self-pollination levels were lowest in the two regions with lowest pollinator service and highest in the region with highest pollinator service. This disagreement between our results and the expectations derived from the reproductive assurance hypothesis may reflect a nonequilibrium situation of the northern H. foetidus populations in relation to their current pollinating environment. PMID- 11410467 TI - Selective seed abortion increases offspring survival in Cynoglossum officinale (Boraginaceae). AB - Selective embryo abortion is one of the evolutionary explanations for the surplus of ovules found in many plant species. To manipulate the level of embryo abortion, we removed ovules and applied nutrients to plants of Cynoglossum officinale (Boraginaceae) after they started to flower. From these two treatments and a control series, seeds were collected, germinated, and transplanted in the field to assess the quality of the resulting offspring. Nutrient addition did not increase the number of seeds per flower significantly. Fewer embryos were aborted in the ovule removal treatment. The seeds produced in the ovule removal treatment had a significantly greater mass and significantly lower survival than the offspring from the control group. This difference in survival indicates that offspring of lower quality are selectively aborted in the control group. Offspring from the nutrient addition treatment survived longer. The offspring of the treatments did not differ significantly from the control group in growth. Simple mathematical calculations, based on the differences in offspring quality that we found, indicate that the selective abortion hypothesis can be an important factor explaining the advantage of the "surplus production" of ovules. PMID- 11410468 TI - Cross-fertility in two tropical tree species: evidence of inbreeding depression within populations and genetic divergence among populations. AB - Knowing the spatial patterns of cross-fertility in natural plant populations yields key insight into biparental inbreeding depression, isolation by distance, and, ultimately, speciation. Three adults each of two tropical tree species (Syzygium rubicundum and Shorea cordifolia) were each crossed with five conspecific pollen donors ranging from self to trees occurring in separate forest reserves (12 and 35 km distance for S. rubicundum and Sh. cordifolia, respectively). Cross-fertility was estimated as fruit set, seed germination, and seedling survivorship and height at 1 yr. Means of most cross-fertility measures increased steadily with outcrossing distance, peaking at 1-2 km for S. rubicundum and 1-10 km for Sh. cordifolia, and then declining at the between-forest crosses. However, seed germination and seedling height for Sh. cordifolia suggested hybrid vigor in between-forest crosses. The mean fitness cost of nearest-neighbor mating relative to crossing with more distant neighbors was 45% for S. rubicundum and 0% for Sh. cordifolia. The mean fitness cost of between-forest crosses was 52% and 70% for the two species. Crossing effects on fitness diminished between the stages of fruit set and 1-yr-old seedlings. Results indicate a strong potential for inbreeding depression within forest tree populations and partial reproductive isolation among forests in Sri Lanka's wet zone. PMID- 11410469 TI - Clinal differentiation and putative hybridization in a contact zone of Pinus ponderosa and P. arizonica (Pinaceae). AB - The widely distributed Pinus subsection Ponderosae is a species complex that has a transition zone among taxa in the southwestern United States. In southern Arizona and New Mexico at least two recognized taxa, Pinus ponderosa var. scopulorum and Pinus arizonica or P. ponderosa var. arizonica, are known to coexist in close proximity. In this study, we report the existence of populations where the taxa are sympatric. One of the key characteristics distinguishing taxa is the number of needles per fascicle; P. ponderosa typically has three, P. arizonica has five. We examined the spatial distribution of needle-number types in a belt transect that covers a transition zone from nearly pure three-needle types at the top of Mount Lemmon to five-needle types downslope, in the Santa Catalina Mountains, Arizona. The spatial distribution is inconsistent with there being both free interbreeding among types and selective neutrality of types. Trees with intermediate types, having combinations of three, four, and five needles and/or mean numbers of needles between 3.0 and 5.0, are spatially concentrated in the middle of the transition zone. The spatial distribution supports the occurrence of hybridization and introgression, and this is consistent with reported crossabilities of the types. The results suggest that selection is acting, either on needle number per se or on other traits of the ecotype with which it may be in linkage disequilibrium, to maintain the observed steep clinal differentiation. PMID- 11410470 TI - Clonality in wild rice (Oryza rufipogon, Poaceae) and its implications for conservation management. AB - Correlations were examined between habitat characters and clonal structures determined by the RAPD (random amplified polymorphic DNA) assay in five populations of Oryza rufipogon in China. Nine of 175 decameric primers were used in the study because they reproducibly amplified polymorphisms. The extent of clonality together with the clonal and sexual reproductive strategies varied greatly among the five populations and correlated with the habitats where they occur. The populations under serious disturbance or seasonal drought tended to have small clones with relatively high clonal diversity caused by sexual reproduction, whereas the populations with little disturbance and sufficient supply of water were prone to have large clones with relatively low clonal variation and low sexual reproduction. Therefore, the dynamics of sexual vs. clonal reproduction of this species depended mainly on environmental factors, such as external disturbance and water supply, rather than latitudes indicated by previous study. These results have important implications for in situ conservation of O. rufipogon. Adequate external disturbance and water supply control are essential for maintaining high clone diversity of in situ conserved populations. According to the extent of clonality of the populations examined, we recommend that an interval of >12 m should be required for collecting samples for ex situ conservation and for population genetic studies to capture possible genetic diversity for O. rufipogon in China. PMID- 11410471 TI - Population structure and genetic diversity of Botrychium pumicola (Ophioglossaceae) based on inter-simple sequence repeats (ISSR). AB - Species of Botrychium reproduce by spores that form subterranean gametophytes and a few, like B. pumicola, also reproduce asexually with subterranean sporophytic gemmae. The goal of this study was to examine the genetic diversity of B. pumicola populations and to better understand the role of gemmae. Ninety-nine individuals from three monitored populations were sampled. The technique of inter simple sequence repeats (ISSR) produced 15 polymorphic loci and identified 71 ISSR genotypes. Sixteen of the ISSR genotypes were shared by more that one individual in a population, representing potential clones. Ten of the 16 shared genotypes were not limited to clusters of plants (groups of plants growing from the same point). The ten potential clones were disjunct (separated by other genotypes) and not in patches as might be expected for an underground propagule. There is a high probability that these shared genotypes arose from independent sexual events suggesting they were not clones. These results suggest that the long-distance dispersal of gemmae is at best a rare event. PMID- 11410472 TI - Nutrient foraging in woodland herbs: a comparison of three species of Uvularia (Liliaceae) with contrasting belowground morphologies. AB - We compared the ability of three closely related species, Uvularia perfoliata, U. sessilifolia, and U. puberula, to forage and explore patches in nutritionally homogeneous and heterogeneous environments. The species differed in type and function of plagiotropic stems and the extent of clonality and physiological integration. Our aim was to determine (1) whether selective placement of roots in high-nutrient patches, i.e., foraging, was accompanied by facilitatory morphological changes such as internode elongation or increased branching, (2) whether foraging ability of species depended on the extent of physiological integration, and (3) how variability in environmental quality influenced the performance of each species. We studied the growth of each species over two seasons in experimental environments. Uvularia perfoliata and U. puberula foraged in high-nutrient patches in heterogeneous environments. Uvularia sessilifolia did not show selective placement of roots. The two clonal species, U. perfoliata and U. sessilifolia, did not show any changes in architectural traits predicted to facilitate foraging. The nonclonal species, U. puberula, was the strongest forager and the most physiologically integrated species, U. sessilifolia, was the weakest forager, in line with the view that physiological integration limits foraging efficiency. Variability in environmental quality had little effect on the performance of the three species. Yield and estimators of fitness were not greater in treatments where more high-quality patches were encountered consecutively than in treatments where fewer high-quality patches were encountered consecutively during growth. PMID- 11410473 TI - Family- and population-level responses to atmospheric CO2 concentration: gas exchange and the allocation of C, N, and biomass in Plantago lanceolata (Plantaginaceae). AB - To ascertain the inheritance of responses to changing atmospheric CO(2) content, we partitioned response to elevated CO(2) in Plantago lanceolata between families and populations in 18 families in two populations. Plants were grown in 35 Pa and 71 Pa partial pressure of CO(2) (pCO(2)) in open-top chambers. We measured above- and belowground mass, carbon (C), nitrogen (N), hexose sugar, and gas exchange properties in both CO(2) treatments. Families within populations differed in mass, mass allocation, root : shoot ratios, aboveground percentage N, C : N ratio, and gas exchange properties. The CO(2) * family interaction is the main indicator of potential evolutionary responses to changing CO(2). Significant CO(2) * family interactions were observed for N content, C : N ratio, and photosynthetic rate (A: instantaneous light-saturated carbon assimilation capacity), intercellular CO(2) concentration, transpiration rate (E), and water use efficiency (WUE = A/E), but not for stomatal conductance. Families differed significantly in acclimation across time. The ratio of A in elevated vs. ambient growth CO(2), when measured at a common internal CO(2) partial pressure was 0.79, indicating down-regulation of A under CO(2) enrichment. Mass, C : N ratio, percentage, C (%C), and soluble sugar all increased significantly but overall %N did not change. Increases in %C and sugar were significant and were coincident with redistribution of N aboveground. The observed variation among populations and families in response to CO(2) is evidence of genetic variation in response and therefore of the potential for novel evolutionary trajectories with rising atmospheric CO(2). PMID- 11410474 TI - Floral movements in response to thunderstorms improve reproductive effort in the alpine species Gentiana algida (Gentianaceae). AB - Studies of floral movements in response to environmental change are rare in the literature, and information about possible adaptive benefits appears nonexistent. The closure of the upright, tubular flowers of alpine gentian (Gentiana algida) were observed during the frequent afternoon thunderstorms characteristic of the central and southern Rocky Mountains (USA). Flowers closed within minutes of an approaching thunderstorm and reopened after direct sunlight returned. Corolla opening widths decreased ~10%/min prior to rainfall, in close correspondence to declines in air and corolla temperatures. Identical floral behavior was also induced experimentally in the field and laboratory by artificial changes in corolla temperature. Corolla closure did not occur during experiments that simulated natural changes in solar irradiance, wind, or absolute humidity during a thunderstorm. Furthermore, individual G. algida plants forced experimentally to remain open during rain had substantial losses of pollen after single rain events (up to 34%) and if forced to remain open for the entire flowering period (59%). Subsequent seasonal reductions in female fitness (up to 73%) also occurred, including seed size and mass, number of ovules produced, number of viable seeds produced per ovule, and seed germination. Thus, corolla closing and opening in G. algida associated with frequent summer thunderstorms may be a behavioral adaptation that improves both paternal and maternal reproductive effort. PMID- 11410475 TI - Habitat-related error in estimating temperatures from leaf margins in a humid tropical forest. AB - Leaf margin characters are strong predictors of mean annual temperature (MAT) in modern plant communities and widely used tools for reconstructing paleoclimates from fossil floras. However, the frequency of nonentire-margined species may vary dramatically between different habitats of the same forest. In this paper we explore the potential for this habitat variation to introduce error into temperature reconstructions, based on field data from a modern lowland forest in Amazonian Ecuador.The data show that the provenance of leaves can influence temperature estimates to an important degree and in a consistent direction. Woody plants growing along lakes and rivers underestimated MAT by 2.5 degrees -5 degrees C, while those in closed-canopy forest provided very accurate predictions. The high proportion of liana species with toothed leaves in lakeside and riverside samples appears to be responsible for a large part of the bias. Samples from closed-canopy forest that included both lianas and trees, however, were more accurate than tree-only or liana-only samples.We conclude that paleotemperature reconstructions based on leaf margin characters will be misleading to the extent that fossilization provides a better record of certain habitats than others. The preponderance of lake and river deposits in the angiosperm fossil record suggests that underestimation of mean annual paleotemperature may be common. PMID- 11410476 TI - Coding and noncoding plastid DNA in palm systematics. AB - Plastid DNA sequences evolve slowly in palms but show that the family is monophyletic and highly divergent relative to other major monocot clades. It is therefore difficult to place the root within the palms because faster evolving, length-variable sequences cannot be aligned with outgroup monocots, and length conserved regions have been thought to give too few characters to resolve basal nodes. To solve this problem, we combined 94 ingroup and 24 outgroup sequences from the length-conserved rbcL gene with ingroup and alignable outgroup sequences from noncoding rps16 intron and trnL-trnF regions. The separate rps16 intron and trnL-trnF region contained about the same number of variable sites (autapomorphies not included) as rbcL, but gave higher retention indices and more clades with bootstrap support. In general, the strict consensus tree based on combined rbcL, rps16 intron, and trnL-trnF data showed more resolution towards the base of the palm family than previous hypotheses of relationships of the Arecaceae. An important result was the position of subfamily Calamoideae as sister to the rest of the palms, but this received <50% bootstrap support. Another result of systematic significance was the indication that subfamily Phytelephantoideae is related to two tribes from subfamily Ceroxyloideae, Cyclospatheae and Ceroxyleae. PMID- 11410477 TI - rbcL data reveal two monophyletic groups of filmy ferns (Filicopsida: Hymenophyllaceae). AB - The "filmy fern" family, Hymenophyllaceae, is traditionally partitioned into two principal genera, Trichomanes s.l. (sensu lato) and Hymenophyllum s.l., based upon sorus shape characters. This basic split in the family has been widely debated this past century and hence was evaluated here by using rbcL nucleotide sequence data in a phylogenetic study of 26 filmy ferns and nine outgroup taxa. Our results confirm the monophyly of the family and provide robust support for two monophyletic groups that correspond to the two classical genera. In addition, we show that some taxa of uncertain affinity, such as the monotypic genera Cardiomanes and Serpyllopsis, and at least one species of Microtrichomanes, are convincingly included within Hymenophyllum s.l. The tubular- or conical-based sorus that typifies Trichomanes s.l. and Cardiomanes, the most basal member of Hymenophyllum s.l., is a plesiomorphic character state for the family. Tubular based sori occurring in other members of Hymenophyllum s.l. are most likely derived independently and more than one time. While rbcL data are able to provide a well-supported phylogenetic estimate within Trichomanes s.l., they are inadequate for resolving relationships within Hymenophyllum s.l., which will require data from additional sources. This disparity in resolution reflects differential rates of evolution for rbcL within Hymenophyllaceae. PMID- 11410478 TI - Phylogenetic relationships within Cornus (Cornaceae) based on 26S rDNA sequences. AB - Phylogenetic relationships within the dogwood genus Cornus have been highly controversial due to the great morphological heterogeneity. Earlier phylogenetic analyses of Cornus using chloroplast DNA (cpDNA) data (including rbcL and matK sequences, as well as restriction sites) and morphological characters suggested incongruent relationships within the genus. The present study generated sequence data from the nuclear gene 26S rDNA for Cornus to test the phylogenetic hypotheses based on cpDNA and morphological data. The 26S rDNA sequence data obtained represent 16 species, 13 from Cornus and three from outgroups, having an aligned length of 3380 bp. Both parsimony and maximum likelihood analyses of these sequences were conducted. Trees resulting from these analyses suggest relationships among subgroups of Cornus consistent with those inferred from cpDNA data. That is, the dwarf dogwood (subg. Arctocrania) and the big-bracted dogwood (subg. Cynoxylon and subg. Syncarpea) clades are sisters, which are, in turn, sister to the cornelian cherries (subg. Cornus and subg. Afrocrania). This red fruited clade is sister to the blue- or white-fruited dogwoods (subg. Mesomora, subg. Kraniopsis, and subg. Yinquania). Within the blue- or white-fruited clade, C. oblonga (subg. Yinquania) is sister to the remainder, and subg. Mesomora is sister to subg. Kraniopsis. These relationships were also suggested by the combined 26S rDNA and cpDNA data, but with higher bootstrap and Bremer support in the combined analysis. The 26S rDNA sequence data of Cornus consist of 12 expansion segments spanning 1034 bp. These expansion segments evolve approximately four times as fast as the conserved core regions. The study provides an example of phylogenetic utility of 26S rDNA sequences below the genus level. PMID- 11410479 TI - Compound pollen cone in a Paleozoic conifer. AB - A rich fossil biota from a Pennsylvanian age deposit of eastern North America contains numerous vegetative and fertile specimens that conform to a single species of primitive walchian conifers. Among the specimens is a compound pollen cone that comprises closely spaced, helically arranged, leaf-like bracts with axillary dwarf shoots. The specimen looks superficially similar to an ultimate vegetative conifer shoot, but there are small appendages in the axil of each bract that represent the fertile dwarf shoots. Dwarf shoots consist of an axis that bears sterile scales and sporophylls with erect pollen sacs. Pollen found in the sacs is monosaccate and conforms to the sporae dispersae genus Potonieisporites Bhardwaj. This cone is a compound shoot system that is morphologically equivalent to the ovulate cones of conifers and to the pollen cones of Paleozoic cordaitaleans and modern gnetophytes. Therefore, it is fundamentally different from the simple pollen cones of other fossil and modern conifers. Discovery of this specimen unexpectedly supports molecular studies that predict a close relationship between Coniferales and Gnetales, and provides fossil evidence to help reconcile the discordant phylogenetic hypotheses of seed plant systematics that have been developed from morphological and molecular data. PMID- 11410480 TI - The battle between tumor suppressors: is gene therapy using p16(INK4a) more efficacious than p53 for treatment of ovarian carcinoma? PMID- 11410481 TI - Approval summary: gemtuzumab ozogamicin in relapsed acute myeloid leukemia. AB - PURPOSE: Gemtuzumab ozogamicin (Mylotarg; Wyeth Laboratories, Philadelphia, PA) consists of a semisynthetic derivative of calicheamicin, a cytotoxic antibiotic linked to a recombinant monoclonal antibody directed against the CD33 antigen present on leukemic myeloblasts in most patients with acute myeloid leukemia (AML). In this study, we review the preclinical and clinical profiles of this immunoconjugate and the regulatory review that led to marketing approval by the United States Food and Drug Administration. EXPERIMENTAL DESIGN: From the literature and manufacturer's data, we review the activity, tolerability, and pharmacokinetics of gemtuzumab ozogamicin in preclinical and Phase I studies and its activity, efficacy, and side effects in three Phase 2 trials of 142 patients with relapsed AML. RESULTS: In Phase I studies, the major toxicity was myelosuppression, especially neutropenia and thrombocytopenia, resulting from the expression of CD33 on myeloid progenitor cells. The Phase 2 dose was 9 mg/m(2) infused i.v. over 4 h, repeated on day 14. A minority of patients experienced acute infusion-related symptoms, usually transient and occasionally requiring hospitalization. The complete response (CR) rate with full recovery of hematopoiesis was 16%. A subset of patients [CRs with incomplete platelet recovery (CRps)] was identified with blast clearance and neutrophil recovery but incomplete platelet recovery. The duration of responses of CRps appeared to be similar to those of the CRs, although the numbers were small. The question of the equivalence of these response groups was a central issue in the review of this new drug application (NDA). After considerable discussion, the Oncology Drugs Advisory Committee recommended allowing inclusion of CRps resulting in an overall response rate in the Phase 2 studies of 30%. In the subgroup of patients over 60 years of age, the overall response rate was 26%. Response duration was difficult to establish because of the high prevalence of postremission therapies. Tolerability and ease of administration may be improved compared with conventional chemotherapy, except for hepatotoxicity, with 31% of patients exhibiting abnormal liver enzymes. One patient died of liver failure in the Phase 2 trials. CONCLUSIONS: Marketing approval of gemtuzumab ozogamicin was granted on May 17, 2000 by the United States Food and Drug Administration under the Accelerated Approval regulations. Gemtuzumab ozogamicin is indicated for the treatment of patients with CD33 positive AML in first relapse who are 60 years of age or older and who are not considered candidates for cytotoxic chemotherapy. The approved dose was 9 mg/m(2) i.v. over 4 h and repeated in 14 days. Completion of the ongoing studies of gemtuzumab ozogamicin in relapsed AML and initiation of randomized clinical trials comparing the effects of gemtuzumab ozogamicin in combination with conventional induction chemotherapy to conventional chemotherapy alone on survival are mandated to confirm clinical benefit under the accelerated approval Subpart H regulations. Postmarketing reports of fatal anaphylaxis, adult respiratory distress syndrome (ARDS), and hepatotoxicity, especially venoocclusive disease (VOD) in patients treated with gemtuzumab ozogamicin, with and without associated hematopoietic stem cell transplantation (HSCT), have required labeling revisions and the initiation of a registration surveillance program. Tumor lysis and ARDS have been reported in patients with leukocytes above 30,000/ml treated with gemtuzumab ozogamicin; therefore, the reduction of leukocyte counts to below 30,000/ml is recommended prior to treatment. Patients should be carefully monitored for acute hypersensitivity, hypoxia, and delayed hepatotoxicity following treatment with gemtuzumab ozogamicin. PMID- 11410482 TI - Induction of topoisomerase II activity after ErbB2 activation is associated with a differential response to breast cancer chemotherapy. AB - ErbB2 (HER-2) gene amplification and overexpression have been shown to predict a better outcome with doxorubicin-based chemotherapy as opposed to alkylator-based chemotherapy in early stage breast cancer. To understand the mechanism of differential response to these two regimens, we have evaluated the effect of signaling through the ErbB2 receptor on downstream enzymes that may affect drug response, using two different models. The first system employs breast cancer cells that have high levels of endogenous ErbB2 by gene amplification (BT-474 and SKBR3 cells). The second system allows us to isolate the effect of ErbB2 receptor mediated intracellular signaling using an epidermal growth factor receptor-ErbB2 chimeric receptor activated by epidermal growth factor. Our experiments show that the cytotoxicity of doxorubicin is inhibited in ErbB2+ breast cancer cells by the anti-ErbB2 antibody, Herceptin. This is accompanied by a decrease in topoisomerase (topo) IIalpha protein and activity, suggesting that this is the mechanism of change in doxorubicin response. In addition, a 10-100-fold (1-2 log) decrease in the LD(50) of doxorubicin is seen after ErbB2 activation using the chimeric receptor model. Furthermore, we see a 100-fold decrease in the LD(50) of etoposide, another topo II inhibitor. This increase in doxorubicin sensitivity is associated with a 4.5-fold increase in the amount of topo IIalpha protein and an increase in topo II activity as measured by DNA decatenating and unknotting activities, as well as cleavable complex formation. In contradistinction to doxorubicin, we have observed an increased resistance to cyclophosphamide chemotherapy after chimeric receptor activation. We propose that the differential benefit seen with doxorubicin- versus alkylator-based chemotherapy in ErbB2+ breast cancer is due, in some cases, to ErbB2-mediated topo IIalpha activation. These data also suggest hypotheses for the optimal sequencing of Herceptin and chemotherapy agents in ErbB2+ breast cancer. PMID- 11410483 TI - Therapy of disseminated B-cell lymphoma xenografts in severe combined immunodeficient mice with an anti-CD74 antibody conjugated with (111)indium, (67)gallium, or (90)yttrium. AB - A radiolabeled antibody (Ab) to CD74 (the MHC class II invariant chain, Ii) was shown previously to effectively kill human B-lymphoma cells in vitro. Conjugates with both Auger electron and beta-particle emitters were able to kill cells, but the former displayed less nonspecific toxicity in the in vitro assay used. In this report, we have extended the studies to an in vivo model of tumor growth. The human B-cell lymphoma Raji was injected i.v. into severe combined immunodeficient mice, and radiolabeled Abs were injected at various times after tumor inoculation. The maximum tolerated dose (MTD), as well as lower doses, was tested. Tumor growth was monitored by hind-leg paralysis. With a 3-5-day interval before Ab injection, anti-CD74 conjugated to either (111)In or (67)Ga, at a dose of 240-350 microCi/mouse, produced a strong therapeutic effect, with greatly delayed tumor growth, and many of the treated mice were tumor free for >6 months. Control mice became paralyzed in 16-24 days, uniformly. Treatment at later time points (9-day interval) had little therapeutic effect. The MTD was required for optimal therapy. With the beta-particle emitter (90)Y, the MTD was much less, 25 microCi/mouse, and at this dose there was only a weak therapeutic effect. In conclusion, the data suggest that low-energy electrons are more effective than beta-particles in this model system. These results may be applicable to humans, particularly in the case of micrometastatic disease. This approach may also be effective with other Abs that accrete in large amounts. PMID- 11410484 TI - Paclitaxel and docetaxel enhance the metabolism of doxorubicin to toxic species in human myocardium. AB - Doxorubicin cardiotoxicity is a multifactorial process in which the alcohol metabolite doxorubicinol mediates the transition from reversible to irreversible damage. We investigated whether the tubulin-active taxane paclitaxel increases conversion of doxorubicin to doxorubicinol, thus explaining the high incidence of congestive heart failure when doxorubicin is used with paclitaxel. Specimens of human myocardium from patients undergoing bypass surgery were processed to obtain cytosolic fractions in which doxorubicin was converted to doxorubicinol by NADPH dependent aldo/keto or carbonyl reductases. In this model, clinically relevant concentrations of paclitaxel (1-2.5 microM) increased doxorubicinol formation by mechanisms consistent with allosteric modulation of the reductases. Stimulation was observed over a broad range of basal enzymatic activity, and was accompanied by a similar pattern of enhanced formation of doxorubicinol aglycone, a metabolite potentially involved in the reversible phase of cardiotoxicity. The closely related analogue docetaxel had effects similar to paclitaxel, but increased doxorubicinol formation over a narrower range of enzymatic activity. The unrelated tubulin-active alkaloid vinorelbine had no effect. These results demonstrate that taxanes have a unique potential for enhancing doxorubicin metabolism to toxic species in human myocardium. The effects on doxorubicinol formation provide clues to explain the clinical pattern of doxorubicin-paclitaxel cardiotoxicity and also caution against the potential toxicity of combining docetaxel with high cumulative doses of doxorubicin. PMID- 11410485 TI - Molecular determination of perivesical and lymph node metastasis after radical cystectomy for urothelial carcinoma of the bladder. AB - PURPOSE: Current methods used to determine the pathological stage of the primary tumor and associated lymphatics after radical cystectomy are tedious, costly, and may lack the sensitivity afforded by molecular approaches such as reverse transcription-PCR (RT-PCR) for markers specific for urothelial tissue such as the uroplakin II (UPII) gene. Thus, we sought to evaluate an objective and sensitive molecular approach for the assessment of perivesical extension and lymph node status after radical cystectomy, based on the detection of UPII expression using RT-PCR and compare this assay to standard clinical and pathological examination. EXPERIMENTAL DESIGN: From November 1999 to September 2000, 27 patients with clinical T(a)-T(3)N(0)M(0) urothelial bladder cancer underwent radical cystectomy, 19 (70%) of which also had pelvic lymphadenectomy. At the completion of cystectomy, systematic biopsies of the external surface of the bladder specimen as well as from the largest palpable lymph node found at lymphadenectomy were obtained for molecular analysis. RT-PCR analysis for UPII mRNA was carried out on these biopsy specimens, and results were compared with data obtained from conventional pathological examination. RESULTS: Pathologically organ-confined tumors had a 42% (5 of 12) incidence of positive signals in the perivesical tissues and 17% (1 of 7) in the lymph nodes. Corresponding percentages for pT(3a)N(0) and pT(3b)-T(4)N(0) lesions were 67% (4 of 6)/25% (1 of 4) and 67% (4 of 6)/33% (2 of 6), respectively. Overall, pathologically node-negative cancers had a perivesical positivity rate of 54% (13 of 24) and a lymph node positivity rate of 25% (4 of 16). All patients with pathologically positive nodes had positive UPII signals in the lymph node sample. CONCLUSIONS: This molecular assay aimed at assessing perivesical extension and lymph node status after radical cystectomy appears to identify patients that may harbor residual disease not appreciated by conventional histology. Larger studies with 5-7-year follow-up will be required to determine the prognostic significance of such molecular information. PMID- 11410486 TI - Persistence of genetically altered fields in head and neck cancer patients: biological and clinical implications. AB - In 1953, Slaughter et al. [D. P. Slaughter et al., Cancer (Phila.), 6: 963-968, 1953] proposed the concept of field cancerization in patients with squamous cell carcinoma of the head and neck (HNSCC) and discussed its clinical significance for the development of second primary tumors and local recurrences. To define the process of field cancerization and its putative clinical implications, we analyzed genetic aberrations in HNSCC and the accompanying macroscopically normal mucosa. In 28 HNSCC patients, loss of heterozygosity was determined in tumor and five noncontiguous mucosal biopsies using eight microsatellite markers at 9p, 3p, and 17p. For patients who showed loss of heterozygosity in their mucosal biopsies, all margins of the surgical specimen were subsequently analyzed to determine the extension of the field. In these cases, additional markers at 8p, 13q, and 18q as well as p53 mutations were included to determine subclonal differences between field and tumor. Genetically altered fields were detected in 36% (10 of 28) of the HNSCC patients. The field varied in size between patients and consisted of genetically different subclones. In 7 of 10 cases, the field extended into the surgical margins. One particular patient with a genetically altered field in a surgical margin developed a local recurrence after 28 months of follow-up. Microsatellite analysis showed that this recurrence had more molecular markers in common with the nonresected premalignant field than with the original tumor, suggesting that this persistent field has progressed further into a new malignancy. Our data show that genetically altered mucosa remains after treatment in a significant proportion of HNSCC patients, which may explain in part the high frequency of local recurrences and second primary tumors. Adequate identification and risk assessment of these genetically altered fields may have profound implications for future patient management. PMID- 11410487 TI - Tumor angiogenesis is associated with MUC1 overexpression and loss of prostate specific antigen expression in prostate cancer. AB - The biological potential of prostate cancer is highly variable and cannot be satisfactorily predicted by histopathological criteria alone. Therefore, additional and more precise information is desirable. Although angiogenesis has been suggested as being of prognostic importance in many human cancers, and MUC1, also known as episialin, was thought to be responsible for the development of metastasis, the role of these parameters in prostate cancer remains unclear. The aim of this study was to investigate whether angiogenesis, assessed as microvessel density (MVD), was correlated with the expression of prostate tumor MUC1 and prostate-specific antigen (PSA) or with histopathological grade at diagnosis, and to determine whether any of these factors might provide additional information with regard to prostate tumor biology. Paraffin-embedded material from 60 patients with prostate carcinoma was examined immunohistochemically, using the monoclonal antibody CD31 to determine MVD, and the monoclonal antibodies CCE831 and ER-PR8 to assess MUC1 and PSA expression, respectively. The tumors were categorized according to the Gleason grading system. MUC1 overexpression was significantly related to a high intratumoral angiogenesis (P = 0.02). By contrast, a high PSA expression by prostate cancer cells was associated with low MVD (P = 0.03). No correlation was found between MUC1 and PSA expression. Usually, high-grade tumors were not PSA-expressive and tended to display increased angiogenesis. These differences, however, were not of statistical significance. Similarly, there was no statistically significant association between histological grade and MUC1 expression or angiogenesis. It is suggested that PSA may have a direct suppressive effect on new blood vessel formation in prostate cancer, whereas the expression of MUC1 in this tumor may be connected with an angiogenic phenotype. Additional studies are obviously needed to clarify the precise role of these proteins in prostate cancer. PMID- 11410488 TI - Detection of genetic alterations in esophageal squamous cell carcinomas and adjacent normal epithelia by comparative DNA fingerprinting using inter-simple sequence repeat PCR. AB - In this study, we screened 19 esophageal squamous cell carcinomas (ESCCs) for the detection of genetic alterations using inter-simple sequence repeat PCR, a DNA fingerprinting approach. Three simple repetitive unanchored primers representing tri- and tetranucleotide repeats [(GTG)(5), (GACA)(4), and (GATA)(4)] were used, and evidence of gains and losses of chromosomal sequences were detected in all tumors (19 of 19 cases) for at least one of the primers. In 13 of these cases, apparently normal marginal epithelia adjacent to the tumors were also collected and examined. Eight of the 13 (62%) patients showed matching somatic mutations in the marginal epithelia adjacent to the tumors. Five of these 8 (63%) marginal epithelial samples were histologically normal, two were dysplastic, and one had extremely rare tumor cells. In 3 of these 13 (23%) cases, the profile bands were also seen to quantitatively increase in intensity, progressing from normal epithelia to marginal epithelia to tumors. Ten profile bands showing gains and one profile band showing loss in tumors compared with the corresponding normal epithelia were cloned, and their origins were determined by sequencing. The DNA sequence of one of the profile bands showing gain in the tumor could be matched to an expressed sequence tag sequence that has been mapped to the 7q22 region, a genomic amplification novel to ESCC. The sequence of the other profile band showing gain in the tumor could be matched to a nonexonic sequence of chromosome 20, whereas the sequences of the remaining profile bands could not be matched with any known sequences after comparison with the genomic sequence data in the European Molecular Biology Laboratory and GenBank databases. The bona fide nature of the gains or losses of 11 profile bands in the original cases was confirmed by direct genomic PCR amplification. The frequencies of these specific gene alterations in tumors were then analyzed in a total of 60 ESCCs, which included 41 additional cases of ESCC. Significantly, 26 of 60 (43%) tumors showed the DNA amplification for the expressed sequence tag sequence of chromosome 7, whereas the frequency of other individual gene alterations ranged from 7% to 15%. It is concluded that the inter-simple sequence repeat PCR strategy is adequate for the detection of somatic mutations in tumors, most of which are quantitative alterations in anonymous genomic sequences. This approach is also suitable for detection of somatic mutations preceding the onset of morphologically detectable neoplasia in ESCC. PMID- 11410489 TI - Fez1/lzts1 alterations in gastric carcinoma. AB - PURPOSE: Loss of heterozygosity (LOH) involving the short arm of chromosome 8 (8p) is a common feature of the malignant progression of human tumors, including gastric cancer. We have cloned and mapped a candidate tumor suppressor gene, FEZ1/LZTS1, to 8p22. Here we have analyzed whether FEZ1/LZTS1 alterations play a role in the development and progression of gastric carcinoma. EXPERIMENTAL DESIGN: We examined Fez1/Lzts1 expression in 8 gastric carcinoma cell lines by Western blot, and in 88 primary gastric carcinomas by immunohistochemistry. Twenty-six of these 88 primary gastric carcinomas were also microdissected and tested for LOH at the FEZ1/LZTS1 locus and for mutation of the FEZ1/LZTS1 gene. Furthermore, we studied the FEZ1/LZTS1 gene regulation and transcriptional control and the methylation status of the 5' region of the gene in all 8 gastric carcinoma cell lines. RESULTS: Fez1/Lzts1 protein was barely detectable in all of the gastric cancer cell lines tested and was absent or significantly reduced in 39 of the 88 (44.3%) gastric carcinomas analyzed by immunohistochemistry, with a significant correlation (P < 0.001) to diffuse histotype. DNA allelotyping analysis showed allelic loss in 3 of 17 (18%) and microsatellite instability in 4 of 17 (23.5%) cases informative for D8S261 at the FEZ1/LZTS1 locus. When we compared the presence of LOH with Fez1/Lzts1 expression, we found loss of protein expression in all three of the tumors with allelic imbalance at D8S261. A missense mutation was detected in one case that did not express Fez1/Lzts1. Hypermethylation of the CpG island flanking the Fez1/Lzts1 promoter was evident in six of the eight cell lines examined as well as in the normal control. CONCLUSIONS: Our findings support FEZ1/LZTS1 as a candidate tumor suppressor gene at 8p in a subtype of gastric cancer and suggest that its inactivation is attributable to several factors including genomic deletion and methylation. PMID- 11410490 TI - Expression of human endogenous retrovirus k envelope transcripts in human breast cancer. AB - PURPOSE: We investigated the expression of human endogenous retroviral (HERV) sequences in breast cancer. EXPERIMENTAL DESIGN: Reverse transcription-PCR (RT PCR) was used to examine expression of the envelope (env) region of ERV3, HERV-E4 1, and HERV-K in breast cancer cell lines, human breast tumor samples, adjacent uninvolved breast tissues, nonmalignant breast tissues, and placenta. Expression of HERV transcripts was confirmed by Northern blot analysis and in situ hybridization (ISH). To evaluate coding potential, amplified HERV sequences were cloned into vectors for expression and sequence analysis. RESULTS: No expression of ERV3 or HERV-E4-1 RNA was detected in the analyzed breast samples. In contrast, HERV-K transcripts were detected in most breast cancer cell lines and many breast tumor tissues. Expression was detected in a small percentage of matched, uninvolved breast tissues and in placentas but not nonmalignant breast tissues. In HERV-K-positive breast cancer tissues, Northern blot analysis demonstrated full-length proviral and spliced env transcripts. ISH demonstrated expression of HERV-K transcripts in breast tumor cells but not in normal or uninvolved breast epithelial cells. Independently isolated clones of HERV-K env cDNA generated recombinant proteins of the expected size. Sequence analysis of env cDNA clones derived from four breast tumor samples revealed >97% identity with the type I HERV-K102, with no premature termination codons. Independent isolates from the same breast tumor sample showed nucleotide sequence differences, suggesting that multiple loci may be transcribed. CONCLUSIONS: These data indicate that HERV-K transcripts with coding potential for the envelope region are expressed frequently in human breast cancer. PMID- 11410491 TI - Radioimmunotherapy with (111)In/(90)Y-2IT-BAD-m170 for metastatic prostate cancer. AB - PURPOSE: Over 31,000 Americans die of androgen-independent metastatic prostate cancer each year. New strategies that do not involve hormonal manipulation but instead recognize the biochemical and molecular characteristics of prostate cancer are needed. Radioimmunotherapy (RIT) uses a tumor-specific monoclonal antibody to deliver systemic, targeted radiation to cancer. The objectives of this Phase I study of (111)In-2IT-BAD-m170 (for imaging) and (90)Y-2IT-BAD-m170 (for therapy) were to determine the toxicity and maximum tolerated dose (MTD), the specificity for targeting metastatic prostate cancer, and the efficacy for palliation of pain. EXPERIMENTAL DESIGN: M170 is a mouse monoclonal antibody that targets adenocarcinomas. Patients with adequate renal and liver function, rising prostate-specific antigen, and androgen-independent metastatic prostate cancer were eligible. After estimation of dosimetry and pharmacokinetics with (111)In 2IT-BAD-m170, a single dose of (90)Y-2IT-BAD-m170 (0.185, 0.370, 0.555, or 0.740 GBq/m(2)) was administered to cohorts of three patients. Pain was assessed objectively by questionnaires before and for 8 weeks after RIT; weekly prostate specific antigen levels were obtained for 2 months after RIT. RESULTS: The MTD of (90)Y-2IT-BAD-m170 was 0.740 GBq/m(2) for patients that had up to 10% of the axial skeleton involved with prostate cancer. Toxicity was almost exclusively confined to reversible myelosuppression. Metastatic prostate cancer was targeted by (111)In-2IT-BAD-m170 in all 17 patients. The mean radiation dose delivered to 39 bone and 18 nodal metastases by (90)Y-2IT-BAD-m170 was 10.5 Gy/GBq (range 2.8 25.1). Thirteen of 17 patients reported pain before (90)Y-2IT-BAD-m170; 7 of these 13 had a partial or complete resolution of pain that lasted an average of 4.3 weeks. CONCLUSIONS: This study determined the MTD of (111)In/(90)Y-2IT-BAD m170 in patients with metastatic prostate cancer. The drugs were well tolerated, targeted metastases, and temporarily palliated pain. PMID- 11410492 TI - Phase I trial of 96-hour continuous infusion of dexrazoxane in patients with advanced malignancies. AB - Dexrazoxane is a bidentate chelator of divalent cations. Pretreatment with short infusions of dexrazoxane prior to bolus doxorubicin has been shown to lessen the incidence and severity of anthracycline-associated cardiac toxicity. However, because of rapid, diffusion-mediated cellular uptake and the short plasma half life of dexrazoxane, combined with prolonged cellular retention of doxorubicin, dexrazoxane may be more effective when administered as a continuous infusion. Thus, a Phase I pharmacokinetic trial of a 96-h infusion of dexrazoxane was performed. Dexrazoxane doses were escalated in cohorts of 3 to 6 patients per dose level. All patients received granulocyte-colony stimulating factor at a dose of 5 microg/kg/day starting 24 h after completion of the dexrazoxane infusion. Plasma samples were collected and analyzed for dexrazoxane by high-performance liquid chromatography. Urine collections were performed at baseline and during the infusion to determine the renal clearance of dexrazoxane and the excretion rate of divalent cations. Twenty-two patients were enrolled at doses ranging from 125 to 250 mg/m(2)/day. Grade 3 and 4 toxicities included grade 4 thrombocytopenia in 2 patients treated at 250 mg/m(2)/day, grade 3 thrombocytopenia and grade 4 nausea and vomiting in 1 patient treated at 221 mg/m(2)/day, grade 4 diarrhea and grade 3 nausea and vomiting in 1 patient treated at 221 mg/m(2)/day, and grade 3 hypertension in 1 patient treated at 166.25 mg/m(2)/day. Steady-state dexrazoxane levels ranged from 496 microg/l (2.2 microM) to 1639 microg/l (7.4 microM). Dexrazoxane plasma CL(ss) and elimination t(1/2) were 7.2 +/- 1.6 l/h/m(2) and 2.0 +/- 0.8 h, respectively. The mean percentage of administered dexrazoxane recovered in the urine at steady state was 30% (range, 10-66%). Urinary iron and zinc excretion during the dexrazoxane infusion increased in 12 of 18 and 19 of 19 patients by a median of 3.7- and 2.4 fold, respectively. These results suggest that dexrazoxane as a 96-h infusion can be safely administered with granulocyte-colony stimulating factor at doses that achieve plasma levels that have been demonstrated previously to inhibit topoisomerase II activity and to induce apoptosis in vitro. Additional studies will be required to determine whether the combination of continuous infusions of dexrazoxane and doxorubicin would provide enhanced cardioprotection compared with the currently recommended bolus or short infusion schedules. PMID- 11410493 TI - Chemotherapy response of breast cancer depends on HER-2 status and anthracycline dose intensity in the neoadjuvant setting. AB - We evaluated the predictive value of a tumor's HER-2 status for chemotherapy response in the neoadjuvant setting and the effect of anthracycline dose intensity on this predictive value. HER-2 status was evaluated by immunochemistry on microbiopsy before neoadjuvant chemotherapy (monoclonal antibody CB-11; Novocastra) in 39 patients (group A) treated with FEC50 (500 mg/m(2) 5 fluorouracil, 50 mg/m(2) epirubicin, and 500 mg/m(2) cyclophosphamide) and 40 patients (group B) treated with FEC100 (500 mg/m(2) 5-fluorouracil, 100 mg/m(2) epirubicin, and 500 mg/m(2) cyclophosphamide). All tumors were stage II or noninflammatory stage III adenocarcinoma. Overall response rate (OR) was evaluated through ultrasound and mammographic measurements. Pathological complete response was evaluated by tumor excision and axillary node resection after six cycles of chemotherapy. Patient and tumor characteristics (age, tumor size, clinical nodal status, SBR grade, hormonal receptor status, and HER-2 expression) were similar in the two groups. In univariate analyses, anthracycline dose was the only factor predictive of response (OR = 61.5% with FEC50; OR = 82.5% with FEC100; P = 0.038). When anthracycline dose was correlated with HER-2 status, an OR of 73.9% was demonstrated in HER-2- tumors (tumors without HER-2 overexpression), and an OR of 12.5% was demonstrated in HER-2+ tumors (tumors with HER-2 with overexpression) in group A. In group B, an OR of 69.5% was demonstrated in HER-2- tumors, and an OR of 100% was demonstrated in HER-2+ tumors. There was no difference in OR for HER-2- tumors treated with FEC50 or FEC100 (P = 0.74). On the other hand, erbB-2+ tumors treated with FEC100 had a significantly better OR than HER-2+ tumors treated with FEC50 (P = 0.0003). In a multivariate analysis, the most powerful predictive factor of OR was a conditional variable associating anthracycline dose with HER-2 status. Low-dose anthracycline and HER-2+ predicted a poor OR, low- or high-dose anthracycline and HER-2- predicted an intermediate OR, and high-dose anthracycline and HER-2+ predicted a high OR. Our results merit additional studies, given the possibility for choosing anthracycline dose according to a tumor's HER-2 status. PMID- 11410494 TI - Survival of tumor cells in stem cell preparations and bone marrow of patients with high-risk or metastatic breast cancer after receiving dose-intensive or high dose chemotherapy. AB - PURPOSE: We evaluated whether dose-intensive or high-dose chemotherapy can eliminate micrometastases in high-risk breast cancer patients. EXPERIMENTAL DESIGN: We monitored cytokeratin (CK)/17-1A positive cells in the bone marrow (BM) and peripheral blood stem cells (PBSC) and studied Her-2/neu serum levels of patients with locally advanced (n = 13; group 1) and metastatic breast cancer (n = 30; group 2) using immunomagnetic separation, immunocytochemistry, and ELISA. RESULTS: CK+ cells were found in the BM of 3 of 13 (23%) group 1 patients before but not after chemotherapy, resulting in an overall survival (OS) of 92% after a median follow-up of 33 months. Contamination of PBSC in 2 of 9 (22%) patients was not associated with decreased survival. In group 2 patients, the CK+ rate was 60% (18 of 30 patients) before and 40% (4 of 10 patients) after therapy with an OS rate of 43% after 29 months. PBSC samples were positive in 7 of 24 (29%) patients. CK+ BM and PBSC led to a rapid progress and short OS, whereas tumor cell-free BM and PBSC resulted in a mean OS of 30 months. The antigen 17-1A was detected on most CK+ cells in both patient groups before therapy, on all of CK+ PBSC, and on CK+ cells in group 2 patients after therapy. Increased Her-2/neu levels were found in group 2 patients before chemotherapy. CONCLUSION: Micrometastatic cells are present in PBSC grafts and can survive even high-dose chemotherapy. The presence of immunotherapeutic target antigens supports the idea that a combined chemoimmunotherapy might be successful in eliminating minimal residual disease. PMID- 11410495 TI - A phase II trial of the cyclin-dependent kinase inhibitor flavopiridol in patients with previously untreated stage IV non-small cell lung cancer. AB - PURPOSE: Flavopiridol is a potent cyclin-dependent kinase inhibitor with preclinical activity against non-small cell lung cancer (NSCLC), inhibiting tumor growth in vitro and in vivo by cytostatic and cytotoxic mechanisms. A Phase II trial was conducted to determine the activity and toxicity of flavopiridol in untreated patients with metastatic NSCLC. EXPERIMENTAL DESIGN: A total of 20 patients were treated with a 72-h continuous infusion of flavopiridol every 14 days at a dose of 50 mg/m(2)/day and a concentration of 0.1-0.2 mg/ml. Dose escalation to 60 mg/m(2)/day was permitted if no significant toxicity occurred. Response was initially assessed after every two infusions; patients treated longer than 8 weeks were then assessed after every four infusions. Plasma levels of flavopiridol were measured daily during the first two infusions to determine steady-state concentrations. RESULTS: This study was designed to evaluate a total of 45 patients in two stages. However, because no objective responses were seen in the first 20 patients, the early-stopping rule was invoked, and patient accrual was halted. In four patients who received eight infusions, progression was documented at 15, 20, 40, and 65 weeks, respectively. The most common toxicities included grade 1 or 2 diarrhea in 11 patients, asthenia in 10 patients, and venous thromboses in 7 patients. The mean +/- SD steady-state concentration of drug during the first infusion was 200 +/- 89.9 nM, sufficient for cytostatic effects in in vitro models. CONCLUSIONS: At the current doses and schedule, flavopiridol does not have cytotoxic activity in NSCLC, although protracted periods of disease stability were observed with an acceptable degree of toxicity. PMID- 11410496 TI - A phase I trial of carboxyamido-triazole and paclitaxel for relapsed solid tumors: potential efficacy of the combination and demonstration of pharmacokinetic interaction. AB - PURPOSE: Preclinical and clinical investigation of the combination of the antiangiogenesis/anti-invasion agent carboxyamido-triazole (CAI) administered with the cytotoxic agent paclitaxel (PAX). EXPERIMENTAL DESIGN: Colony-forming assays were used to test the activity of CAI plus PAX on A2780 human ovarian cancer. The sequence of CAI followed by PAX (CAI>Pax) was modeled in nude mice to test for potential additive toxicity. The Phase I clinical dose escalation schema tested p.o. administered CAI in PEG-400 (50-100 mg/m(2)) or micronized CAI (250 mg/m(2)) for 8 days followed by a 3-h infusion of PAX (110-250 mg/m(2)) every 21 days. Patients were assessed for toxicity, pharmacokinetics of CAI and PAX, and disease outcome. RESULTS: In preclinical studies, CAI>Pax was additive in A2780 human ovarian cancer cell lines when CAI (1 or 5 microM) preceded subtherapeutic doses of PAX. CAI did not reverse PAX resistance and collateral resistance to CAI was documented in PAX-resistant cells. CAI>PAX administration had no overt additive toxicity in nude mice. Thirty-nine patients were treated on a dose escalation Phase I trial using daily oral CAI for 8 days followed by the PAX infusion. Pharmacokinetic analysis revealed that PAX caused an acute increase in circulating CAI concentrations in a dose-dependent fashion. No additive or cumulative toxicity was observed, and grade 3 nonhematological toxicity was rare. Three partial responses and two minor responses were observed. CONCLUSIONS: The sequential combination of CAI and PAX is well tolerated, and the activity observed suggests that further study of the combination is warranted. PMID- 11410497 TI - The P-glycoprotein antagonist PSC 833 increases the plasma concentrations of 6alpha-hydroxypaclitaxel, a major metabolite of paclitaxel. AB - PURPOSE: Overexpression of P-glycoprotein (Pgp) is one mechanism of drug resistance in cancer chemotherapy. A Phase I trial was conducted using PSC 833, a Pgp antagonist, in combination with paclitaxel in patients with refractory cancer. The objective of this study was to assess the effect of PSC 833 on the metabolism of paclitaxel and characterize the differences in 6alpha hydroxypaclitaxel pharmacokinetics. In addition, we examined the possibility of enhanced cytotoxicity of paclitaxel by the coexistence of 6alpha hydroxypaclitaxel. EXPERIMENTAL DESIGN: Patients received paclitaxel 35 mg/m(2)/day by continuous intravenous infusion (CIVI) x 4 days without PSC 833 in cycle 1 and escalating doses of paclitaxel (13.1, 17.5, or 21.3 mg/m(2)/day CIVI x 4 days) with 5 mg/kg PSC 833 by mouth every 6 h x 7 days in cycle 2. Plasma samples were analyzed for both paclitaxel and its major metabolite with high performance liquid chromatography methods. Using human liver microsomes, we studied the effect of PSC 833 on the metabolism of paclitaxel. In addition, the in vitro cytotoxicity of 6alpha-hydroxypaclitaxel alone and in combination with paclitaxel was evaluated. RESULTS: Twenty-one of 22 patients had a metabolite peak (6alpha-hydroxypaclitaxel) observed in the chromatogram of plasma samples from cycle 2 when they received paclitaxel in combination with PSC 833. This metabolite was not detectable in plasma obtained during the first cycle when they received paclitaxel without PSC 833. During cycle 2, the mean concentrations of 6alpha-hydroxypaclitaxel and paclitaxel were 0.10 +/- 0.074 and 0.079 +/- 0.041 microg/ml, respectively. A moderate association was observed between total bilirubin and 6alpha-hydroxypaclitaxel concentrations (P = 0.015, r = 0.52; n = 21). Human liver microsome experiments showed that a PSC 833 concentration as high as 10 microM did not affect the production of 6alpha-hydroxypaclitaxel. Paclitaxel cytotoxicity in HL60 and K562 human leukemia cells was increased in the presence of noncytotoxic concentrations of 6alpha-hydroxypaclitaxel. CONCLUSIONS: PSC 833 increases the plasma concentration of 6alpha hydroxypaclitaxel during paclitaxel therapy. Inhibition of cytochrome P-450 3A4 by PSC 833 may explain this in part, although other mechanisms cannot be excluded. PMID- 11410498 TI - Defective transforming growth factor beta signaling pathway in head and neck squamous cell carcinoma as evidenced by the lack of expression of activated Smad2. AB - PURPOSE: Transforming growth factor beta (TGF-beta) regulates cell growth and differentiation, in normal squamous epithelium, via specific TGF-beta receptors and intracellular signaling molecules (Smads). We have previously observed that TGF-beta type II receptor (TbetaR-II) expression decreases in squamous cell carcinomas as tumors become less differentiated and more biologically aggressive. However, a small fraction of tumors remain TbetaR-II positive. In this article, we examine the integrity of the other members of the TGF-beta-signaling machinery, the Smad proteins. EXPERIMENTAL DESIGN: Thirteen archived head and neck squamous cell carcinomas were selected from the files of the Pathology Department of the H. Lee Moffitt Cancer Center. Protein immunoexpression was quantitated by image analysis in the context of histopathological parameters. Mutation analysis of the MADR2/Smad2 gene was also performed. RESULTS: In both TbetaR-II-positive and TbetaR-II-negative tumors, expression of the non-TGF-beta specific Smads (4, 6, and 7) was variable, whereas expression of the pathway specific Smad2 was lost in 38% of the tumors. Expression of the activated, phosphorylated form of this molecule, Smad2-P, was lost in approximately 70% of the tumors. No abnormal mRNA expression and no mutations in the MADR2/Smad2 gene were observed. CONCLUSIONS: These results suggest that multiple defects in TGF beta signaling, both at the receptor and postreceptor level, may play a role in the oncogenesis of head and neck squamous cell carcinoma. PMID- 11410499 TI - Tumor angiogenesis and its possible role in intravasation of colorectal epithelial cells. AB - PURPOSE: To determine whether an increase in tumor angiogenesis facilitates intravasation of colorectal epithelial cells, we compared intratumoral microvessel counts with the presence of circulating colorectal epithelial cells in the portal venous blood from patients with colorectal carcinomas. EXPERIMENTAL DESIGN: Circulating colorectal epithelial cells were detected by a reverse transcription-PCR assay to amplify guanylyl cyclase C (GCC) transcripts. The extent of tumor vascularization was quantitatively assessed by immunohistochemical staining with anti-CD31 antibody. RESULTS: Colorectal epithelial cells (as measured by GCC mRNA expression) were detected in the portal venous blood in 30 of 58 patients (52%). The mean (+/- SD) microvessel count in the tumors from patients with expression of GCC mRNA in their portal venous blood was 82.74 +/- 24.97. The corresponding values in the tumors from patients without expression of GCC mRNA in portal venous blood was 65.96 +/- 19. For each 10 microvessel increase per x200 field, the risk of colorectal epithelial cell presence in the portal venous blood increased 1.52-fold (95% confidence interval, 1.19-2.12; P = 0.005). CONCLUSION: High intratumoral vessel count was noted to be a valuable factor for predicting the presence of colorectal epithelial cells in the portal venous blood. PMID- 11410500 TI - Elevated serum levels of type I collagen degradation marker ICTP and tissue inhibitor of metalloproteinase (TIMP) 1 are associated with poor prognosis in lung cancer. AB - PURPOSE: The aim of this study was to evaluate the correlation between type I collagen degradation marker ICTP, MMP (matrix metalloproteinase)-9, and tissue inhibitor of metalloproteinase (TIMP)-1 and to compare their value as prognostic factors in lung cancer. EXPERIMENTAL DESIGN: From the sera of 141 lung cancer patients, we assessed markers of type I collagen synthesis (PINP and PICP) and degradation (ICTP) by radioimmunoassays, and we assessed MMP-9 and its tissue inhibitor TIMP-1 by ELISA. There were 62 squamous cell carcinomas, 42 adenocarcinomas, 14 small cell carcinomas, and 23 cases with other histology. Seventeen of these patients had advanced disease. Sixty-seven patients had been operated on, 33 had received radiation therapy, 7 had received chemotherapy, and the rest had received other treatment combinations. RESULTS: We examined the relationship between these markers and found a correlation between ICTP and MMP-9 (r = 0.201; P = 0.01) or TIMP-1 (r = 0.415; P = 0.00). Elevated serum concentrations of ICTP (>5 microg/liter) and/or TIMP-1 (>300 ng/ml) correlated with poor prognosis. In univariate regression analysis, ICTP had prognostic value (odds ratio, 1.6462; P < 0.03): the patients with elevated serum concentrations of ICTP (>5 microg/liter) had a 64% higher risk of dying from lung cancer than did patients with opposite values. CONCLUSIONS: Our results indicated that ICTP and TIMP-1 are good prognostic markers in lung cancer. The association between serum MMP-9 and ICTP suggests that MMP-9 could play a role in the degradation of the extracellular matrix producing ICTP in this pathological situation. PMID- 11410501 TI - Diagnostic strategy for analytical scanning of BRCA1 gene by fluorescence assisted mismatch analysis using large, bifluorescently labeled amplicons. AB - The aim of this study was to develop a protocol for reliable, sensitive, and cost effective mutation scanning of the BRCA1 gene, based on a modification of fluorescence-assisted mismatch analysis. The main features of this method are: (a) robust PCR amplification and strandspecific labeling of 25 large amplicons using uniform conditions and universal fluorescent primers; and (b) sensitive characterization of the position of sequence changes. The diagnostic accuracy of this method was tested by scanning the large exon 11 in 12 DNA samples with reported mutations. In a blind test, specific patterns of fluorescence profiles were obtained, and all were attributed correctly, without sequencing, to each mutation or polymorphism. Seven breast/ovarian cancer families with high probability of BRCA1-related predisposition were screened. Three truncating mutations (of which one was novel and three were missense changes, including two novel ones) were detected. The three missense mutations affect the highly conserved BRCT domain. Scanning by FAMA appears to be free of biases for particular types of sequence changes-except for exon deletions/duplications, which cannot be detected by conventional PCR-based methods-and allows substantial savings in the number of sequencing reactions and in the time invested in their interpretation. Therefore, it lends itself to screening structurally complex loci in the diagnostic context and in other fields of genetic analysis. PMID- 11410502 TI - Prediction of peritoneal micrometastasis by peritoneal lavaged cytology and reverse transcriptase-polymerase chain reaction for matrix metalloproteinase-7 mRNA. AB - PURPOSE: Peritoneal dissemination is the most common cause of death associated with gastric cancer. In this study, we report the significance of molecular diagnosis of peritoneal dissemination by means of matrix metalloproteinase-7 (MMP 7) reverse transcriptase-PCR (RT-PCR) assay using preoperative peritoneal wash fluid. EXPERIMENTAL DESIGN: Preoperative peritoneal lavage by paracentesis was performed on 152 patients with gastric cancer. The peritoneal lavaged fluid was subjected to RT-PCR analysis with primers specific for MMP-7 and conventional cytological Papanicolaou examination. RESULTS: The MMP-7 RT-PCR assay was able to detect cancer cells at densities even lower than 10 cells/sample. There was no signal of MMP-7 mRNA from mesothelial cells, fibroblasts, peripheral blood, and lavaged fluid from patients with benign disease. Cytological examination and MMP 7 RT-PCR assay results were positive for 27 (18%) and 28 (18%) samples, respectively. The sensitivity for the prediction of peritoneal dissemination by cytology and MMP-7 RT-PCR assay were 46% and 33%, but the combination analysis using both parameters improved the sensitivity rate with 62%. Logistic regression analysis revealed that the cytological examination and MMP-7 RT-PCR assay are independent predictors of peritoneal dissemination. CONCLUSION: The combination of cytological examination and RT-PCR assay of preoperative peritoneal lavaged fluid is a highly efficient and reliable method for the selection of patients for adjuvant i.p. chemotherapy. PMID- 11410503 TI - The expression of the urokinase plasminogen activator system in metastatic murine osteosarcoma: an in vivo mouse model. AB - The role of urokinase plasminogen activator (uPA) in osteosarcoma is poorly understood. We examined the importance of uPA, its receptor, uPAR, and its inhibitor, PAI-1, in our in vivo model of metastatic osteosarcoma. Rodent osteosarcoma cells (UMR 106-01) were inoculated into the tibia of athymic mice. Animals were sacrificed and autopsied at 4 days to 5 weeks after inoculation. Tibiae and lungs were excised, fixed, and examined histologically and by in situ hybridization. Osteosarcoma development was associated with tibial swelling and lameness, and radiographic changes included osteolysis and new bone formation. Lung metastases developed spontaneously. In the tibial tumors, uPAR mRNA was expressed early (4 days), whereas uPA and PAI-1 mRNA increased as the tumor invaded the surrounding tissue (3 weeks). There was also an increase in the mRNA expression of the osteoblast-related genes, alpha1(I) procollagen and osteopontin, but not matrix Gla protein. Lung metastases also expressed mRNA for the uPA system and the bone-related proteins. We have produced a model of metastatic osteosarcoma, which typifies the characteristics of the human tumor. Our results suggest that the uPA system plays a role in the local aggressiveness and metastasis of osteosarcoma and, in particular, indicates a possible therapeutic role for uPAR antagonists in the treatment of osteosarcoma. PMID- 11410504 TI - Expression of hypoxia-inducible factor 1alpha in epithelial ovarian tumors: its impact on prognosis and on response to chemotherapy. AB - PURPOSE: To investigate the impact of expression of hypoxia-inducible factor (HIF)-1alpha on prognosis and on response to chemotherapy in epithelial ovarian tumors. EXPERIMENTAL DESIGN: Expression of HIF-1alpha protein was studied by immunohistochemistry in 102 specimens of epithelial ovarian cancers, in 50 borderline tumors, and in 20 cystadenomas. Results were correlated with p53, p21, and bcl-2 expression, microvessel density (MVD), apoptotic rate of tumor cells, and survival. RESULTS: In 68.6% of ovarian cancers and 88% of borderline tumors, expression of HIF-1alpha was observed. There was a significant correlation of HIF 1alpha protein expression and MVD (P < 0.001). HIF-1alpha overexpression alone and MVD showed no impact on survival of cancer patients. Furthermore, the response to platinum-based chemotherapy was independent from HIF-1alpha expression. Expression of HIF-1alpha correlated with apoptotic rate in the majority of cases, especially in low malignant potential tumors. In contrast, in cancer patients with strong expression of HIF-1alpha and p53 protein overexpression, not only a significantly increased MVD (P = 0.032, Mann-Whitney test) but also a significantly shorter overall survival was observed (P < 0.0001, Cox regression). The apoptotic rate was very low in these tumors. CONCLUSIONS: HIF-1alpha protein overexpression alone has no impact on the prognosis of ovarian cancer. The combination of HIF-1alpha protein overexpression with nonfunctional p53, however, indicates a dismal prognosis. PMID- 11410505 TI - Evaluation of the United States Food and Drug Administration-approved scoring and test system of HER-2 protein expression in breast cancer. AB - PURPOSE: The purpose of this study was to investigate the prognostic significance of assessment of human epidermal growth factor receptor (HER)-2 oncogene protein overexpression of breast cancer tissue by the United States Food and Drug Administration (FDA)-approved HercepTest and grading system (negative, 0 or 1+; weakly positive, 2+; strongly positive, 3+). Furthermore, results of the HercepTest were correlated with immunohistochemical results obtained using different antibodies and protocols and with HER-2 oncogene gene amplification assessed by fluorescence in situ hybridization (FISH). EXPERIMENTAL DESIGN: HER-2 status in 303 patients with lymph node-positive breast cancer was investigated by using a rabbit polyclonal antibody (DAKO) by conventional immunohistochemistry and by applying the HercepTest. Furthermore, the monoclonal antibody CB-11 was used in conventional immunohistochemistry and with the NexES automatic stainer, which is also under consideration for FDA approval for determination of eligibility for Herceptin therapy. Results were compared with FISH analysis performed in all 2+ and 3+ specimens (103 cases) and 104 HER-2-negative specimens. RESULTS: 3+ positive carcinomas were found in 8.9-15.7% of specimens. FISH revealed that almost exclusively 3+ positive cases were amplified, with the HercepTest and the NexES automatic stainer giving the best results. In univariate analysis, staining with the HercepTest revealed a significantly worse prognosis in 3+ cases. Also, 3+ cases were significantly associated with lower estrogen receptor levels and histological grade III tumors. CONCLUSIONS: This study shows that the results of the FDA-approved HER-2 grading and test system correlated strongly with findings in FISH. Furthermore, HercepTest proved to be of prognostic relevance. Strict adherence to the given protocols is critical. PMID- 11410506 TI - Alterations of the 9p21 and 9q33 chromosomal bands in clinical bladder cancer specimens by fluorescence in situ hybridization. AB - PURPOSE: To better define cytogenetic mechanisms of CDKN2 loss at 9p21 and of DBCCR1 loss at 9q33 in bladder cancer, and to determine correlation with p53 and pRb. EXPERIMENTAL DESIGN: Two-color fluorescence in situ hybridization (FISH) using a chromosome 9 centromeric probe and locus-specific probes was performed. p53 and pRb were assessed by immunohistochemistry. RESULTS: Thirty-seven of fifty five (67%) samples exhibited 9p21 loss, and 32 of 44 (73%) exhibited 9q33 loss. Twelve of 43 informative samples exhibited only 9p21 loss (5 cases) or only 9q33 loss (7 cases). Homozygous deletions were noted at 9p21 and 9q33 in 31 and 14% of cases, respectively, but 9q33 homozygous deletions were generally observed in only a minor clone. There was no correlation of any chromosome 9 loss with stage, but stage did correlate with chromosome 9 ploidy status; aneusomy 9 was observed in 33% of T(a) lesions and 71% of more advanced cases (P = 0.01). Aneusomy 9 was loosely correlated with p53 abnormalities (P = 0.07), but no correlation between any chromosome 9 and pRb abnormalities was discerned. CONCLUSIONS: This study strengthens the proposition that chromosome 9 losses occur early in bladder oncogenesis and before p53 alterations or development of aneusomy. The correlation of aneusomy 9 with p53 abnormalities is consistent with the presumed role of p53 in maintaining cytogenetic stability. Although the observed homozygous deletions strengthen the hypotheses that CDKN2 and DBCCR1 are important tumor suppressor genes, there is no evidence that either is a more critical or an earlier target for oncogenesis. PMID- 11410507 TI - Loss of H-cadherin protein expression in human non-small cell lung cancer is associated with tumorigenicity. AB - Abnormalities in the H-cadherin gene have been described in several human cancers. However, their biological significance remains undetermined. To investigate the role of H-cadherin in non-small cell lung cancer (NSCLC), a chimera H-cadherin-green fluorescent protein (GFP) expressed in Cos-7 cells was used to identify an anti-H- cadherin antibody, HCD-1. Western blot analysis was performed in six NSCLC cell lines and 35 pairs of primary NSCLC tumors and nonmalignant lung tissue obtained from surgical resections using HCD-1. Loss of H cadherin expression was seen in five (83%) of the six NSCLC cell lines, whereas loss of E-cadherin was seen in three (50%) of the six. H-cadherin expression was lost in 15 (43%) of 35 NSCLC surgical tumor specimens, whereas E-cadherin expression was lost in 6 (17%) of 35. H-cadherin was expressed in all of the nonmalignant lung tissue from all of the surgical specimens. Fourteen of 35 tumors were heterotransplanted s.c. in nude mice. Tumorigenicity in nude mice was associated with both loss of H-cadherin expression (P = 0.03) and loss of E cadherin expression (P = 0.05). Loss of H-cadherin was also associated with a more advanced local tumor growth, although the difference was not significant. The results indicate that loss of H-cadherin is frequent in human NSCLC and suggest that it facilitates the implantation and local growth of human NSCLC tumors. PMID- 11410509 TI - Quantitation of marrow disease in neuroblastoma by real-time reverse transcription-PCR. AB - PURPOSE: GD2 is abundantly expressed in neuroblastoma (NB). GD2 synthesis is dependent on key enzyme beta 1,4-N-acetylgalactosaminyltransferase (GD2 synthase). We explore the potential of GD2 synthase mRNA as a molecular marker of minimal residual disease by first comparing it quantitatively with immunocytology and then testing its clinical utility. EXPERIMENTAL DESIGN: A real-time reverse transcription-PCR assay to quantify mRNA of GD2 synthase was developed. Quantitation was normalized to endogenous control glyceraldehyde-3-phosphate dehydrogenase in a multiplex PCR. RESULTS: The upper limit of normal was defined by 31 normal marrow and blood samples, achieving a sensitivity of one NB cell in 10(6) normal mononuclear cells. When 155 bone marrows from 100 NB patients were studied, GD2 synthase mRNA levels correlated well with the number of GD2-positive cells, as measured by immunocytology using anti-GD2 antibodies (r = 0.96). This is the first demonstration of the quantitative relationship between a specific mRNA and the actual number of tumor cells. In a pilot study, the level of this transcript in sequential marrow samples of five stage 4 NB patients correlated closely with their clinical status. At 24 months after diagnosis, available remission bone marrows from patients with advanced NB diagnosed at >1 year of age initially treated with protocols N6 and N7 at Memorial Sloan-Kettering Cancer Center (n = 44) were analyzed for GD2 synthase mRNA. Positivity was strongly associated with progression-free (P < 0.005) and overall survival (P < 0.001). CONCLUSIONS: Measurement of tumor cells by real-time quantitative reverse transcription-PCR of GD2 synthase has potential clinical utility, especially for the detection of minimal residual disease. PMID- 11410508 TI - Nucleotide substitution in the ectodomain of trail receptor DR4 is associated with lung cancer and head and neck cancer. AB - Allelic loss of chromosome 8p21-22 occurs frequently in cancer, including lung and head and neck squamous cell cancer. The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptors, including proapoptotic DR4 and KILLER/DR5, are located on 8p21-22. TRAIL receptors are candidate tumor suppressor genes, because their inactivation would be expected to result in deficient apoptotic signaling. To investigate the involvement of DR4 in human cancer, we have determined the genomic structure of DR4 and screened 31 lung cancer cell lines [14 small cell lung cancer and 17 non-small cell lung cancer (NSCLC)], many with deletions at 8p21-22, and 21 primary NSCLC samples for mutations in DR4. We found two missense alterations in the ectodomain of DR4. One, at nucleotide 626, changes a cytosine to a guanine (C626G) and results in a substitution of an arginine for threonine. The other, at nucleotide 422, changes a guanine to adenine (G422A) and results in a substitution of a histidine for arginine. Using genomic DNA sequencing and RFLP analysis, we show that these two alterations cosegregated in 96% of all of the samples (n = 243) evaluated (tumor and normal). The frequency of being homozygous for both altered alleles was 35% in the lung cancer cell lines but only 13% in age- and race-matched controls, which was a significant increase (chi(2) = 5.2, P = 0.023). The frequency of homozygosity for both alleles was also significantly increased in the primary NSCLC samples (chi(2) = 9.2, P = 0.002) as compared with the age- and race matched controls. To determine whether the altered alleles are specific for lung cancer, we evaluated 19 head and neck squamous cell cancer and 25 gastric adenocarcinoma samples. Forty-seven % of the former and 44% of the latter were homozygous for both the C626G and G422A alterations, and this was significantly elevated relative to age- and race-matched controls (chi(2) = 8.6, P = 0.003 and chi(2) = 8.2, P = 0.004). These alterations result in amino acid changes in or near the ligand-binding domain of DR4 and, based on the crystal structure of DR5 and its homology with DR4, have the potential to affect TRAIL binding to DR4. Our results suggest that the altered DR4 alleles may be associated with, and should be investigated additionally as potential markers for, predisposition to common malignancies. PMID- 11410510 TI - Expression pattern of fatty acid-binding proteins in human normal and cancer prostate cells and tissues. AB - PURPOSE: Fatty acid-binding protein (FABP) expression patterns were evaluated as potential markers and therapeutic targets for prostate cancer. EXPERIMENTAL DESIGN: FABP expression levels were determined by reverse transcription-PCR in cultured prostate normal and tumor cells and in human biopsy samples. Regulation of cellular processes was examined using FABP antisense constructs. RESULTS: Prostate cells express a variety of different FABPs. Liver (L)- and intestine FABPs were elevated 5-9-fold in prostate cancer compared with normal primary prostate cells. In contrast, adipose- and epidermal-FABPs were markedly down regulated (3-20-fold) in cancer versus normal cells. Similar expression patterns were found in human tissue biopsy samples. However, brain-FABP had a distinct pattern of expression: it was overexpressed only in LNCaP cells and in well differentiated tissue samples, suggesting a stage-specific expression profile. Secretion of L-FABP protein was observed from DU 145 prostate cancer cells, but not in the culture fluid of normal prostate epithelial cells. Antisense oligodeoxynucleotides, designed to block production of epidermal-FABP (a marker for normal prostate cells), caused increased proliferation in DU 145 prostate cancer cells. In vivid contrast, antisense oligodeoxynucleotides to L-FABP (overexpressed in prostate cancer) decreased proliferation and caused apoptosis. CONCLUSIONS: We propose that there is a distinct balance between these groups of FABPs, whose altered regulation in cells may play a role in prostate cancer. Furthermore, the pattern of expression and secretion of FABPs have the potential to serve as a diagnostic marker for an aggressive phenotype of prostate cancer. PMID- 11410511 TI - Measures of cell turnover (proliferation and apoptosis) and their association with survival in breast cancer. AB - Our objective was to investigate the prognostic significance of cell turnover (apoptosis and proliferation) in breast cancer patients. Apoptosis was microscopically quantitated on histological sections from 791 breast cancer patients with long-term follow-up (median, 16.3 years). Apoptotic counts were also compared with proliferation data (mitotic counts and MIB-1 labeling); apoptosis data derived from terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end labeling (TUNEL) assay; and pathobiological variables, including p53, erbB-2, and estrogen receptor (ER). High apoptotic counts were associated with increased cellular proliferation, ER negativity, immunopositivity of erbB-2 and p53 (P < 0.0001), and shortened disease-specific survival (DSS; P = 0.0009) and disease-free survival (DFS; P = 0.0006). Other factors associated with shortened DFS and DSS by univariate analysis were high tumor grade, nodal metastases, and large tumor size (P < 0.0001 for each). Multivariate analysis of data for all of the patients demonstrated that tumor size, nodal status, ER, histological grade, and erbB-2 showed independent prognostic value. In node negative patients, tumor size and mitotic rate per 1000 cells independently predicted DFS (P = 0.0055). Tumor grade was the only independent predictor of DSS. For node-positive patients, tumor size, nodal status, ER, and erbB-2 were independent prognostic factors. The number of mitoses per 1000 was independently associated with DFS (P = 0.043) but not with DSS. Apoptosis data did not provide independent prognostic value in any, node-positive or node-negative, breast cancer patients. PMID- 11410512 TI - Frequent death-associated protein kinase promoter hypermethylation in multiple myeloma. AB - Death-associated protein (DAP) kinase is a novel gene regulating apoptosis induced by IFN-gamma. In B-cell malignancies, loss of DAP kinase expression is commonly associated with promoter hypermethylation. These characteristics of DAP kinase may be of particular relevance in multiple myeloma (MM), a B-lineage malignancy in which prolonged survival capacity of the malignant plasma cells may be critical in the induction and maintenance of tumor cells. PURPOSE: The involvement and potential role of DAP kinase in MM pathogenesis was examined. EXPERIMENTAL DESIGN: In this investigation, methylation-specific PCR was conducted on primary MM and MM cell lines. Methylation status findings were correlated with clinical parameters. RESULTS: We first demonstrated frequent DAP kinase hypermethylation in 24 of 36 primary MMs (20 of 26 at diagnosis and 4 of 10 with relapse/residual MM after treatment), 1 of 2 patients with monoclonal gammopathy of undetermined significance, and 1 of 3 MM cell lines studied. The high frequency of DAP kinase hypermethylation was similarly observed in MM of different stages, immunoglobulin isotypes, and histological grades, with or without plasmacytomas. Although not statistically significant, the overall survival of patients with DAP kinase methylation was notably shortened among 23 MM patients followed prospectively (P = 0.38 by Kaplan-Meier method and log-rank test). This preliminary finding suggests prognostic implications of DAP kinase in MM that may deserve further investigation. CONCLUSIONS: Our data suggest an important role for DAP kinase in MM tumorigenesis. PMID- 11410513 TI - Clinicopathological and molecular evidence indicating the independence of bronchioloalveolar components from other subtypes of human peripheral lung adenocarcinoma. AB - Although human lung adenocarcinoma has diverse histological subtypes, the correlation between histological subtypes and occurrence of the p53 gene mutation has been given less attention. We investigated 145 surgically resected lung adenocarcinomas to search for the incidence of p53 mutations and for record data on survival in each histological subtype, according to the new WHO criteria (1999). The frequency of p53 mutation in bronchioloalveolar carcinoma (BAC; 0% in 17 cases) and BAC with invasive growth component (BAC-invasive; 11% in 27 cases), which is conventionally categorized as the mixed subtype in WHO typing, were apparently significantly lower than in other types (non-BAC including acinar, papillary, solid, or mixed histology with these subtypes; 48% in 101 cases; P < 0.01). Multivariate analysis revealed that the histological subtype including BAC invasive was a strong, independent, and significant prognostic factor (P < 0.03), as were tumor size and pathological stage (P < 0.001 and 0.002, respectively) for overall survival. However, the occurrence of p53 mutation itself was seen to be significant only in case of the univariate analysis. Therefore, histological subtyping may be a better prognostic indicator than is p53 mutation. These findings suggest that the WHO classification with the BAC and BAC-invasive from other histological subtypes may prove useful to predict the outcome for surgically treated patients with lung adenocarcinoma. PMID- 11410514 TI - Pathological features and BRCA1 mutation screening in premenopausal breast cancer patients. AB - PURPOSE: Risk calculations for carrying BRCA1/BRCA2 mutations are based on family history and the age of onset of cancers. However, women may carry these deleterious mutations without a strong family history. Additional criteria for risk estimation would be of value. It has been recently established that BRCA1 associated breast cancers are associated with poor tumor differentiation (TD3) and estrogen receptor (ER) negativity. The aim of this study is to determine whether morphological features of breast cancers in premenopausal patients (age < 45 years) could determine additional women who may benefit from BRCA1 screening. EXPERIMENTAL DESIGN: In a prospective, systematic study of 76 consecutive breast cancer patients (age < 45 years), genomic DNA was obtained from peripheral blood, and eight mutations in BRCA1 (10.5%) were found. Archival paraffin-embedded breast cancer specimens were then analyzed for tumor differentiation and ER status. RESULTS: In patients < 45 years of age, 25% (6 of 24) of ER-negative and TD3 breast cancers were found to harbor mutations in BRCA1. Only 5.6% (2 of 36) of BRCA1-associated breast cancers did not have this morphological profile, compared with 94.4% (34 of 36) patients without BRCA1 mutations, giving an odds ratio of 5.67 (95% confidence interval, 1.04-32; P = 0.05). Finally, only one patient with BRCA1 mutations had a significant family history. CONCLUSIONS: In patients with early-onset breast cancer, the use of morphological criteria provides an additional strategy to determine those patients who might benefit from genetic testing. PMID- 11410515 TI - Association of urokinase-type plasminogen activator and its inhibitor with disease progression and prognosis in ovarian cancer. AB - PURPOSE: Urokinase-type plasminogen activator (uPA) and its inhibitor, plasminogen activator inhibitor (PAI)-1, have been shown to be related to poor prognosis in a variety of malignant solid tumors. Studies on the prognostic relevance of uPA and PAI-1 in ovarian cancer, however, have been inconclusive. The current study tests the hypothesis that elevated expression of uPA and PAI-1 is associated with prognosis and disease progression. EXPERIMENTAL DESIGN: uPA and PAI-1 were prospectively measured by quantitative ELISA in tumor samples from 103 ovarian cancer patients (82 primary invasive epithelial carcinomas, 9 low malignant potential tumors, and 12 recurrent ovarian carcinomas). RESULTS: uPA but not PAI-1 levels were consistently associated with malignant progression, with levels increased from low malignant potential tumors to primary tumors (uPA, P = 0.04; PAI-1, P = 0.019), from early to advanced disease stages (uPA, P = 0.014; PAI-1, P = 0.23), and from primary to intra-abdominal metastatic tumors (uPA, P = 0.001; PAI-1, P = 0.16). High uPA and PAI-1 levels were associated with residual tumor volumes of >1 cm (P = 0.001 and P = 0.016, respectively). Among invasive International Federation of Gynecologists and Obstetrician stages I-IV tumors, elevated levels of uPA (>5.5 ng/mg) and PAI-I (>18.8 ng/ml) were associated with a shortened progression-free survival (uPA, P = 0.003; PAI-1, P = 0.039) and overall survival (uPA, P = 0.0002; PAI-1, P = 0.007). In multivariate analysis, uPA retained prognostic independence for progression-free survival (P = 0.037) and overall survival (P = 0.006). CONCLUSIONS: These data suggest that the uPA/PAI-1 axis may play an important role in the intra-abdominal spread and reimplantation of ovarian cancer cells. The prognostic relevance of uPA and PAI-1 supports their possible role in the malignant progression of ovarian cancer. PMID- 11410516 TI - Widespread expression of thioredoxin and thioredoxin reductase in non-small cell lung carcinoma. AB - We investigated the expression of thioredoxin (Trx) and thioredoxin reductase (TrxR) in 89 non-small cell lung carcinomas. Additionally, immortalized human bronchial epithelial cells (BEAS 2B) and four human lung carcinoma cells lines (A549, SK-MES-1, CALU-6, and A427) were studied by Western blot and reverse transcription-PCR for the synthesis of Trx and TrxR protein and mRNA expression in vitro. The histological samples were also studied for immunohistochemical p53 and proliferating cell nuclear antigen expression and apoptosis. In non neoplastic lung, Trx and TrxR expression was seen in bronchial epithelial cells and alveolar macrophages, metaplastic alveolar epithelial cells, and chondrocytes of the bronchus. In non-small cell lung carcinomas, there was a widespread expression of Trx and TrxR with only three and eight cases negative, respectively. Trx and TrxR expression was located in both cytoplasmic and nuclear compartments of the cells. There was a statistical association between cytoplasmic and nuclear Trx or TrxR expression. Grade I-II tumors showed stronger cytoplasmic and nuclear Trx and TrxR immunoreactivity than grade III tumors. No association was found between p53 and proliferating cell nuclear antigen expression and Trx or TrxR immunoreactivity. However, apoptosis was inversely associated with nuclear Trx and TrxR positivity. In the cell lines studied, both non-neoplastic BEAS 2B cells and all of the carcinoma cell lines expressed Trx and TrxR proteins and mRNA. The results show that these redox-regulating proteins are highly expressed in lung carcinomas taking part in activation of transcriptional factors and regulation of apoptosis in non-small cell lung carcinoma. In high-grade tumors, Trx and TrxR expression is diminished, suggesting loss of redox regulation in tumors with low differentiation. PMID- 11410517 TI - Biochemical correlates of mTOR inhibition by the rapamycin ester CCI-779 and tumor growth inhibition. AB - The rapamycin ester, CCI-779, potently inhibits cell growth in vitro, inhibits tumor growth in vivo, and is currently in Phase I clinical trials. To further understand the relationship between plasma systemic exposure and inhibition of the target Ser/Thr kinase, mTOR/FRAP, two assays have been developed. The first assay involves determination of the 4E suppressor protein (4E-BP1) bound to eukaryotic initiation factor 4E (eIF4E), and the second is direct Western analysis of phosphorylation of residue Thr(70) of 4E-BP1. Under normal growth conditions in vitro, rapamycin caused rapid association of 4E-BP1 with eIF4E within 1 h in Rh30 and GC(3) human tumor cells. Association was persistent up to 16 h. In mice, administration of rapamycin (5 or 20 mg/kg) caused rapid association of 4E-BP1 with eIF4E within 4 h in both human colon adenocarcinoma GC(3) and rhabdomyosarcoma Rh30 xenografts. Using phospho-specific antibody against Thr(70) of 4E-BP1, rapid and persistent dephosphorylation within 30 min of exposure to rapamycin was detected in Rh18 rhabdomyosarcoma cells. Evaluation of CCI-779 against Rh18 xenografts showed this tumor to be growth inhibited at daily dose levels of > or =8.7 mg/kg. Because immunoblotting may be more suitable for assaying tumor biopsy tissue, a "blinded" comparison between the effect of CCI-779 on Thr(70) phosphorylation and growth inhibition of human tumor xenografts was undertaken. Mice were treated daily for 5 days with CCI-779 (20 mg/kg/day) or with drug vehicle, and tumor diameters were measured. Tumors were excised 1 h after the final administration and frozen, and phospho Thr(70) was determined by Western blot analysis. The correlation coefficient for decreases in Thr(70) phosphorylation and growth inhibition was high (r(2), 0.99). The results indicate that an assay of decreases in phosphorylation of Thr(70) of 4E-BP1 may be a useful surrogate for determining the inhibition of mTOR activity in tumor specimens. PMID- 11410518 TI - In vitro and in vivo adenovirus-mediated p53 and p16 tumor suppressor therapy in ovarian cancer. AB - PURPOSE: The objectives of this study were to determine the effects of adenovirus mediated p16 and p53 on growth and apoptosis in ovarian cancer cells and on survival in nude mice implanted with human ovarian cancer cells. EXPERIMENTAL DESIGN: SKOV-3 ip1 (p53 and p16 null), 2774 (p53 and p16 mutant), and OVCA 420 (p53 and p16 wild-type) cells were used for in vitro studies. SKOV-3 ip1, 2774, and Hey A8 (p53 and p16 wild-type) cells were used in the nude mouse studies. The E1-deleted adenoviruses containing p53, p16, or beta-galactosidase cDNA were transfected into the different cell types or inoculated into the nude mice after injection with ovarian cancer cells. RESULTS: Cell counting, microtetrazolium, and anchorage-independent growth assays on transfected cells demonstrated that p16 and the p16/p53 combination suppressed growth, whereas p53 did not (except in the anchorage-independent growth assay). Although cells infected with the p16/p53 combination had decreased growth compared with cells infected with either tumor suppressor alone, the difference was only statistically significant compared with p53. p16, p53, and the p16/p53 combination all increased apoptosis in the cells. In the nude mice, p16 treatment resulted in the longest survival for all three models, although it only reached statistical significance for the 2774 and SKOV-3 ip1 groups. CONCLUSIONS: Overall, p16 demonstrated greater growth inhibition than p53 both in vivo and in vitro. The p16/p53 combination demonstrated a consistent trend toward increased growth suppression and apoptosis over p16 or p53 alone. Adenovirus-mediated p16 may be a viable future treatment for ovarian cancer. PMID- 11410519 TI - Interleukin-6 induces androgen responsiveness in prostate cancer cells through up regulation of androgen receptor expression. AB - Interleukin-6 (IL-6) induces prostate cancer (CaP) cell proliferation in vitro. Several lines of evidence suggest that IL-6 may promote CaP progression through induction of an androgen response. In this work, we explored whether IL-6 induces androgen responsiveness through modulation of androgen receptor (AR) expression. We found that in the absence of androgen, IL-6 increased prostate-specific antigen (PSA) mRNA levels and activated several androgen-responsive promoters, but not the non-androgen responsive promoters in LNCaP cells. Bicalutamide, an antiandrogen, abolished the IL-6 effect and IL-6 could not activate the PSA and murine mammary tumor virus reporters in AR-negative DU-145 and PC3 cells. These data indicate the IL-6 induces an androgen response in CaP cells through the AR. Pretreatment of LNCaP cells with SB202190, PD98059, or tyrphostin AG879 [p38 mitogen-activated protein kinase (MAPK), MAP/extracellular signal-regulated protein kinase kinase 1/2, and ErbB2 MAPK inhibitors, respectively) but not wortmannin (PI3-kinase inhibitor) blocked IL-6-mediated induction of the PSA promoter, which demonstrates that IL-6 activity is dependent on a MAPK pathway. Finally, IL-6 activated the AR gene promoter, resulting in increased AR mRNA and protein levels in LNCaP cells. These results demonstrate that IL-6 induces AR expression and are the first report of cytokine-mediated induction of the AR promoter. Taken together, our results suggest that IL-6 induces AR activity through both increasing AR gene expression and activating the AR in the absence of androgen in CaP cells. These results provide a mechanism through which IL-6 may contribute to the development of androgen-independent CaP. PMID- 11410520 TI - Differential sensitivity of normal and malignant breast epithelial cells to genistein is partly mediated by p21(WAF1). AB - Genistein, a soy metabolite, is a potential chemopreventive agent against various types of cancer. There are several studies documenting molecular alterations leading to cell cycle arrest and induction of apoptosis in a variety of cancer cells; however, no studies, to date, have shown the effect of genistein in isogenic normal and malignant breast epithelial cells. In this study, we investigated whether genistein shows any differential sensitivity to normal (MCF10A and MCF12A) and malignant (MCF10CA1a and MDA-MB-231) breast epithelial cells. We found that genistein causes a greater degree of G(2)-M arrest and induces apoptosis in malignant cell lines compared with normal breast epithelial cells. After genistein treatment, flow cytometric analysis revealed a hyperdiploid population in malignant cells that was not observed in normal cells. Cell cycle regulator p21(WAF1), which is known to be up-regulated by genistein treatment, was greatly induced at RNA and protein levels in normal cells, whereas its level was only slightly induced in malignant MDA-MB-231 cells and not detectable in malignant MCF10CA1a cells. Therefore, we investigated the causal role of p21(WAF1) in the differential sensitivity of genistein among these cell lines. We examined the effects of genistein on p21(WAF1) -/- and p21(WAF1) +/+ HCT116 cells, which were used as controls prior to studies on breast cancer cells. We found that there was a greater degree of cell cycle arrest and apoptosis in p21(WAF1) -/- cells compared with p21(WAF1) +/+ HCT116 cells after genistein treatment. Flow cytometric analysis after genistein treatment showed a significant number of p21(WAF1) -/- cells in the hyperdiploid population, which are probably programmed to die through apoptotic processes. To further confirm the causal role of p21(WAF1) in genistein-mediated cell cycle arrest and apoptosis, we down-regulated p21(WAF1) by antisense p21(WAF1) cDNA transfection experiments. We found that both normal and malignant p21(WAF1) antisense (AS) expressing clones became more sensitive to G(2)-M arrest after genistein treatment. Flow cytometric analysis showed an increase in the hyperdiploid population in the AS clones. Further evaluation showed an increase in apoptosis in malignant AS clones but not in normal breast epithelial AS clones. These results suggest that p21(WAF1) may play an important role in determining the sensitivity of normal and malignant breast epithelial cells to genistein. PMID- 11410521 TI - Inhibition of insulin-like growth factor I receptor increases the antitumor activity of doxorubicin and vincristine against Ewing's sarcoma cells. AB - Innovative treatment modalities are needed for Ewing's sarcoma (ES), a neoplasm with a disappointingly low survival rate despite the use of aggressive multimodal therapeutic approaches. We and others (D. Yee et al., J. Clin. Investig., 86: 1806-1814, 1990; K. Scotlandi et al., Cancer Res., 56: 4570-4574, 1996) have previously shown the existence and the pathogenetic relevance of an autocrine loop, mediated by the insulin-like growth factor-I receptor (IGF-IR), which is crucial for survival and proliferation of ES cells in vitro. Moreover, we reported that the IGF-IR-blocking monoclonal antibody (MAb), alphaIR3, as well as suramin, a drug that can interfere with growth factor by binding to the receptors, inhibited both the tumorigenic and the metastatic ability of ES cells in athymic mice. In this study, we analyzed whether agents that can block the IGF IR-mediated loop are of value in association with conventional cytotoxic drugs for the design of more effective therapeutic regimens. Both alphaIR3 MAb and suramin treatment significantly increased the antitumor in vitro effects of doxorubicin and vincristine, two drugs with a leader action on ES. These findings were obtained by both simultaneous and sequential treatments. Analysis of the proliferation rate and of apoptosis revealed that alphaIR3 MAb and suramin significantly enhanced the G(1)-phase rate induced by doxorubicin, without substantially affecting doxorubicin-G(2)-M-blockage of cell cycle, and significantly increased the induction of apoptosis, which confirmed that the specific blockage of IGF-IR deprives ES cells of an important tool for the prevention of drug-induced apoptosis. Moreover, combination treatments of doxorubicin plus alphaIR3 MAb significantly increase the doxorubicin-induced impairment of the ability of ES cells to form colonies in soft agar. In conclusion, we showed that, in ES, the blockage of IGF-IR by a neutralizing MAb or by suramin may greatly potentiate the antitumor activity of conventional chemotherapeutic drugs. PMID- 11410522 TI - Multidrug resistance proteins MRP3, MRP1, and MRP2 in lung cancer: correlation of protein levels with drug response and messenger RNA levels. AB - Previously (L. C. Young et al., Clin. Cancer Res., 5: 673-680, 1999), we found, in a panel of 23 lung cancer cell lines that had not been selected for in vitro drug resistance, that the mRNA levels of MRP3 and MRP1, two members of the ATP binding cassette superfamily of transport proteins, correlated with resistance to doxorubicin, vincristine, VP-16, and cis-diamminedicholoroplatinum(II). To extend these studies, we measured multidrug resistance protein (MRP)1, MRP2, and MRP3 protein levels in a panel of 30 lung cancer cell lines that included the original 23 cell lines as well as an additional 7 unselected lung cancer cell lines. In the case of MRP3, a polyclonal antibody was developed that was found to be a sensitive reagent for the detection of MRP3 by Western blot analysis. We found good agreement in the original 23 cell lines between the cognate mRNA and protein levels for MRP1, MRP2, and, especially, MRP3 (r, 0.852), supporting the use of semiquantitative PCR to predict MRP1, MRP2, and MRP3 protein levels in patient samples. There were also strong correlations between the mRNA and protein levels of MRP3 and MRP1, which suggested that these genes might be expressed in a coordinate manner. MRP3, MRP1, and MRP2 protein levels were higher in the non small cell lung cancer (NSCLC) than in the SCLC cell lines and, in addition, MRP3 and MRP2 were detected almost exclusively in the NSCLC cell lines. Finally, we found that both MRP3 and MRP1, but not MRP2, protein levels correlated with decreased sensitivity of these lung cancer cell lines to doxorubicin, VCR, VP-16, and cis-diamminedicholoroplatinum(II). These findings are consistent with our hypothesis that both MRP3 and MRP1 are components of the multifactorial multidrug resistance phenotype of lung cancer and that MRP3 contributes to the intrinsic resistance of NSCLC cells. PMID- 11410523 TI - Carboxymethyl benzylamide dextran and tamoxifen combination inhibits tumor growth and angiogenesis. AB - We showed previously that a carboxymethyl dextran benzylamide (CMDB7) blocks angiogenesis of MDA-MB-435 carcinoma and its lung metastases in nude mice. In this study, we examined the combination effects of CMDB7 and tamoxifen (TAM) on cell proliferation, tumor growth, and angiogenesis on the MCF-7RAS cells. We showed that CMDB7 and TAM acted in a synergistic manner to inhibit the growth of MCF-7RAS cells, blocking them in G(0)/G(1) phase of the cell cycle. For 7 weeks, the CMDB7- (300 mg/kg/week) and TAM- (20 mg/kg/week) treated groups showed tumor growth inhibition of about 66% and 76%, respectively. Combined treatments with CMDB7 and TAM block the tumor development by 94% and induce a complete regression of 4 of 8 mice. Histological analysis showed markedly less neovascularization (88%) in the tumors treated with a combination of CMDB7 and TAM. This antiangiogenic activity was further demonstrated by direct inhibition of endothelial cell proliferation. Overall, this study points to the potential use of a combination of CMDB7 and TAM to inhibit tumor angiogenesis that can prevent tumor progression. PMID- 11410524 TI - IL (interleukin)-1alpha promotes nuclear factor-kappaB and AP-1-induced IL-8 expression, cell survival, and proliferation in head and neck squamous cell carcinomas. AB - Interleukin 1alpha (IL-1alpha) is an important regulatory cytokine, the release of which after an injury can induce activation of transcription factors nuclear factor (NF)kappaB and activator protein (AP-1), which promote expression of genes involved in cell survival, proliferation, and angiogenesis. IL-1alpha is expressed autonomously by head and neck squamous cell carcinomas (HNSCCs) and a variety of other cancers, raising the possibility that IL-1alpha may serve as an autocrine factor that stimulates the activation of prosurvival transcription factors and target genes in cancer. In this study, we examined the role of IL 1alpha in the activation of NFkappaB and AP-1, the expression of proangiogenic cytokine IL-8, and in the survival and proliferation of HNSCC cell lines. HNSCCs were found to secrete and respond to functional IL-1alpha, in that culture supernatant from a high IL-1alpha-secreting line, UM-SCC-11B, could induce secretion of cytokine IL-8 by a low IL-1alpha-secreting line, UM-SCC-9; and the induction of IL-8 secretion could be blocked by the anti-IL-1alpha-neutralizing antibody or the IL-1 receptor antagonist (IL-1RA). Furthermore, IL-1alpha could induce the expression of IL-8 through an autocrine mechanism, in that transfection of UM-SCC-9 cells with a plasmid encoding IL-1alpha resulted in the increased coexpression of IL-1alpha and IL-8; whereas transfection with a plasmid encoding IL-1RA lacking the secretory leader sequence led to the decreased coexpression of IL-1alpha and IL-8. IL-1alpha was found to induce coexpression of IL-8 through the activation of NFkappaB and AP-1, in that mutation of the NFkappaB site within the IL-8 promoter abolished autocrine- and recombinant IL 1alpha-induced IL-8 reporter gene activity, whereas mutation in AP-1 partially decreased IL-8 reporter gene activity in UM-SCC-9 cells. Intracellular expression of IL-1RA decreased NFkappaB reporter gene activity, indicating that endogenously expressed IL-1alpha contributes to constitutive NFkappaB activation in this HNSCC line. Expression of IL-1alpha affected survival of UM-SCC-9, inasmuch as transfection of cells with plasmid encoding IL-1alpha or IL-1RA led to the increased or decreased survival of cells cotransfected with a beta-galactosidase reporter gene, respectively. IL-1alpha was also found to promote the increased growth of UM-SCC-9 cells in vitro. We demonstrate that exogenous and endogenous IL-1alpha contributes to the transcriptional activation of NFkappaB and AP-1, to the expression of IL-8, and to cell survival and the growth of HNSCC in vitro. PMID- 11410525 TI - Preferential induction of apoptosis by interferon (IFN)-beta compared with IFN alpha2: correlation with TRAIL/Apo2L induction in melanoma cell lines. AB - On the basis of in vitro inhibition of tumor cell growth, IFNs have been generally considered to be antiproliferative proteins. To probe further the potential mechanisms of the antitumor effects of IFNs, we have assessed apoptosis in response to IFN-alpha2 and IFN-beta in cell lines of varied histologies, with a focus on melanomas. Many of the cell lines tested underwent apoptosis in response to IFN-beta, as assessed both by Annexin V and terminal deoxynucleotidyl transferase-mediated nick end labeling staining. In general, IFN-beta had greater growth inhibitory and proapoptotic effects than IFN-alpha2 on all cell lines. The melanoma cell line WM9, sensitive to growth inhibition by IFNs, had a greater degree of apoptosis than A375 melanoma cells, which were largely resistant to antigrowth effects of IFNs. IFN-beta-induced apoptosis was dependent on activation of the caspase cascade with cleavage of caspases 3, 8, and 9 and of the caspase 3 substrate, poly(ADP-ribose) polymerase. Caspase inhibitors benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl keton or benzyloxycarbonyl-Asp-Glu-Val Asp-fluoromethyl keton, inhibited IFN-beta-induced apoptosis. Other changes associated with apoptosis, including the movement of cytochrome c from mitochondria to cytoplasm and DNA fragmentation, were also identified in response to IFN-beta. Apo2L ligand [tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)] was one of the early genes induced by IFN-beta in apoptosis sensitive WM9 cells. Other sensitive melanoma cell lines had a similar IFN-beta specific induction of TRAIL. Neutralizing antibody to TRAIL inhibited IFN-beta induced apoptosis in WM9 cells. In resistant A375 cells, IFN-beta did not induce TRAIL/Apo2L expression. Thus, induction of TRAIL by IFNs in some tumor types may initiate the apoptotic cascade. This study offers another mechanism for the antitumor effects of IFNs. PMID- 11410526 TI - Changing colors in mice: an inducible system that delivers. PMID- 11410527 TI - The languages of bacteria. PMID- 11410528 TI - BH3-only proteins that bind pro-survival Bcl-2 family members fail to induce apoptosis in the absence of Bax and Bak. AB - The BH3-only proteins Bim and Bad bind to the antiapoptotic Bcl-2 proteins and induce apoptosis in wild-type cells and cells from either bax(-/-) or bak(-/-) animals. In contrast, constitutively active forms of Bim and Bad failed to induce apoptosis in bax(-/-)bak(-/-) cells. Expression of Bax restored susceptibility of the cells to Bim and Bad. In addition, Bax but not Bim or Bad sensitized the bax( /-)bak(-/-) cells to a wide variety of cell death stimuli including UV irradiation, chemotherapeutic agents, and ER stress. These results suggest that neither activation of BH3-only proteins nor suppression of pro-survival Bcl-2 proteins is sufficient to kill cells in the absence of both Bax and Bak. Furthermore, whereas mouse embryo fibroblasts (MEF) expressing only Bax or Bak displayed resistance to transformation, bax(-/-)bak(-/-) MEF were nearly as prone to oncogenic transformation as p53(-/-) MEF. Thus, the function of either Bax or Bak appears required to initiate most forms of apoptosis and to suppress oncogenic transformation. PMID- 11410529 TI - A crucial component of the endoderm formation pathway, CASANOVA, is encoded by a novel sox-related gene. AB - casanova (cas) mutant zebrafish embryos lack endoderm and develop cardia bifida. In a substractive screen for Nodal-responsive genes, we isolated an HMG box containing gene, 10J3, which is expressed in the endoderm. The cas phenotype is rescued by overexpression of 10J3 and can be mimicked by 10J3-directed morpholinos. Furthermore, we identified a mutation within 10J3 coding sequence that cosegregates with the cas phenotype, clearly demonstrating that cas is encoded by 10J3. Epistasis experiments are consistent with an instructive role for cas in endoderm formation downstream of Nodal signals and upstream of sox17. In the absence of cas activity, endoderm progenitors differentiate into mesodermal derivatives. Thus, cas is an HMG box-containing gene involved in the fate decision between endoderm and mesoderm that acts downstream of Nodal signals. PMID- 11410530 TI - casanova encodes a novel Sox-related protein necessary and sufficient for early endoderm formation in zebrafish. AB - Early endoderm formation in zebrafish requires at least three loci that function downstream of Nodal signaling but upstream of the early endodermal marker sox17: bonnie and clyde (bon), faust (fau), and casanova (cas). cas mutants show the most severe phenotype as they do not form any gut tissue and lack all sox17 expression. Activation of the Nodal signaling pathway or overexpression of Bon or Fau/Gata5 fails to restore any sox17 expression in cas mutants, demonstrating that cas plays a central role in endoderm formation. Here we show that cas encodes a novel member of the Sox family of transcription factors. Initial cas expression appears in the dorsal yolk syncytial layer (YSL) in the early blastula, and is independent of Nodal signaling. In contrast, endodermal expression of cas, which begins in the late blastula, is regulated by Nodal signaling. Cas is a potent inducer of sox17 expression in wild-type embryos as well as in bon and fau/gata5 mutants. Cas is also a potent inducer of sox17 expression in MZoep mutants, which cannot respond to Nodal signaling. In addition, ectopic expression of cas in presumptive mesodermal cells leads to their transfating into endoderm. Altogether, these data indicate that Cas is the principal transcriptional effector of Nodal signaling during zebrafish endoderm formation. PMID- 11410531 TI - The lac operator-repressor system is functional in the mouse. AB - We report the successful transfer of a fully functional lac operator-repressor gene regulatory system to the mouse. The key component is a lac repressor transgene that resembles a typical mammalian gene both in codon usage and structure and expresses functional levels of repressor protein in the animal. We used the repressor to regulate the expression of a mammalian reporter gene consisting of the tyrosinase promoter embedded with three short lac operator sequences and the tyrosinase coding sequence. Pigmentation of the mouse was controlled by the interaction of the lac repressor with the regulatable Tyrosinase transgene in a manner that was fully reversible by the lactose analog IPTG. Direct control of mammalian promoters by the lac repressor provides tight, reversible regulation, predictable levels of de-repressed expression, and the promise of reversible control of the endogenous genome. PMID- 11410532 TI - Terminal proteins essential for the replication of linear plasmids and chromosomes in Streptomyces. AB - Linear plasmids and chromosomes of the bacterial genus Streptomyces have proteins of unknown characteristics and function linked covalently to their 5' DNA termini. We purified protein attached to the end of the pSLA2 linear plasmid of Streptomyces rochei, determined the N-terminal amino acid sequence, and used this information to clone corresponding genes from a S. rochei cosmid library. Three separate terminal protein genes (here designated as tpgR1, tpgR2, and tpgR3), which map to the S. rochei chromosome and to 100-kb and 206-kb linear plasmids contained in S. rochei, were isolated and found to encode a family of similar but distinct 21-kD proteins. Using tpgR1 to probe a genomic DNA library of Streptomyces lividans ZX7, whose linear chromosome can undergo transition to a circular form, we isolated a S. lividans chromosomal gene (tpgL) that we found specifies a protein closely related to, and functionally interchangeable with, TpgR proteins for pSLA2 maintenance in S. lividans. Mutation of tpgL precluded propagation of the pSLA2 plasmid in a linear form and also prevented propagation of S. lividans cells that contain linear, but not circular, chromosomes, indicating a specific and essential role for tpg genes in linear DNA replication. Surprisingly, Tpg proteins were observed to contain a reverse transcriptase-like domain rather than sequences in common with proteins that attach covalently to the termini of linear DNA replicons. PMID- 11410533 TI - The 19S complex of the proteasome regulates nucleotide excision repair in yeast. AB - Previous studies suggest that the amino-terminal ubiquitin-like (ubl) domain of Rad23 protein can recruit the proteasome for a stimulatory role during nucleotide excision repair in the yeast Saccharomyces cerevisiae. In this report, we show that the 19S regulatory complex of the yeast proteasome can affect nucleotide excision repair independently of Rad23 protein. Strains with mutations in 19S regulatory subunits (but not 20S subunits) of the proteasome promote partial recovery of nucleotide excision repair in vivo in rad23 deletion mutants, but not in other nucleotide excision repair-defective strains tested. In addition, a strain that expresses a temperature-degradable ATPase subunit of the 19S regulatory complex manifests a dramatically increased rate of nucleotide excision repair in vivo. These data indicate that the 19S regulatory complex of the 26S proteasome can negatively regulate the rate of nucleotide excision repair in yeast and suggest that Rad23 protein not only recruits the 19S regulatory complex, but also can mediate functional interactions between the 19S regulatory complex and the nucleotide excision repair machinery. The 19S regulatory complex of the yeast proteasome functions in nucleotide excision repair independent of proteolysis. PMID- 11410534 TI - Drosophila Fos mediates ERK and JNK signals via distinct phosphorylation sites. AB - During Drosophila development Fos acts downstream from the JNK pathway. Here we show that it can also mediate ERK signaling in wing vein formation and photoreceptor differentiation. Drosophila JNK and ERK phosphorylate D-Fos with overlapping, but distinct, patterns. Analysis of flies expressing phosphorylation site point mutants of D-Fos revealed that the transcription factor responds differentially to JNK and ERK signals. Mutations in the phosphorylation sites for JNK interfere specifically with the biological effects of JNK activation, whereas mutations in ERK phosphorylation sites affect responses to the EGF receptor-Ras ERK pathway. These results indicate that the distinction between ERK and JNK signals can be made at the level of D-Fos, and that different pathway-specific phosphorylated forms of the protein can elicit different responses. PMID- 11410535 TI - Specific tracheal migration is mediated by complementary expression of cell surface proteins. AB - Migration of the Drosophila tracheal cells relies on cues provided by nearby cells; however, little is known about how these signals specify a migratory path. Here we investigate the role of cell surface proteins in the definition of such a pathway. We have found that the PS1 integrin is required in the tracheal cells of the visceral branch, whereas the PS2 integrin is required in the visceral mesoderm; both integrins are necessary for the spreading of the visceral branch over its substratum. This is the first identification of a cell surface molecule with expression restricted to a subset of tracheal cells that all migrate in a given direction. We have also found that expression of PS1 in the visceral branch is regulated by the genes that direct tracheal cell migration, showing that integrin expression is part of the cell-fate program that they specify. These results support a model in which signal transduction determines the tracheal migratory pathways by regulating the expression of cell surface proteins, which in turn interact with surface molecules on the surrounding cell population. PMID- 11410536 TI - The u-boot mutation identifies a Hedgehog-regulated myogenic switch for fiber type diversification in the zebrafish embryo. AB - Developmental programs that govern the embryonic diversification of distinct kinds of muscles in vertebrates remain obscure. For instance, the most widely recognized attribute of early diversity among skeletal myoblasts is their ability to differentiate exclusively into fibers with slow or fast contractile properties. However, we know little about the developmental basis and genetic regulation of this seminal event in vertebrate myogenesis. Here we show that in the zebrafish, the u-boot gene acts as a myogenic switch that regulates the choice of myoblasts to adopt slow versus fast fiber developmental pathways. In u boot mutant embryos, slow muscle precursors abort their developmental program, failing to activate expression of the homeobox gene prox1 and transfating into muscle cells with fast fiber properties. Using oligonucleotide-mediated translational inhibition, we have investigated the role of prox1 in this program. We find that it functions in the terminal step of the u-boot controlled slow fiber developmental pathway in the regulation of slow myofibril assembly. Our findings provide new insight into the genetic control of slow versus fast fiber specification and differentiation and indicate that dedicated developmental pathways exist in vertebrates for the elaboration of distinct elements of embryonic muscle pattern. PMID- 11410537 TI - Control of axillary bud initiation and shoot architecture in Arabidopsis through the SUPERSHOOT gene. AB - The aerial architecture of flowering plants is determined to a large extent by shoot growth and shoot branching arising from the initiation and growth of axillary meristems. We have identified an Arabidopsis mutant, supershoot (sps), which is characterized by a massive overproliferation of shoots, such that a single plant can generate 500 or more inflorescences. Analysis of the mutant plants shows that the primary defect is because of an increase in the number of meristems formed in leaf axils, together with release of bud arrest, resulting in reiterative branch formation from rosette and cauline leaves. The SPS gene is shown here to encode a cytochrome P450, and together with a 3- to 9-fold increase in levels of Z-type cytokinins in sps mutant plants, indicate a role for SPS in modulating hormone levels. The expression pattern of SPS, with strong expression at the leaf axils, correlates well with the phenotypic defects. Our results indicate that control of shoot branching in Arabidopsis may be accomplished in part by suppression of axillary meristem initiation and growth through the localized attenuation of cytokinin levels at sites of bud initiation. PMID- 11410538 TI - Soundings: a psychological equivalent of medical percussion. PMID- 11410540 TI - Depression and sickle cell disease. AB - Depressive symptoms are very common in patients with a chronic medical illness such as sickle cell disease (SCD). Clinicians may fail to recognize depression in such patients when, as in SCD, the two conditions have many overlapping symptoms. Based on a Medline database search, we review papers addressing the relationship between depression and SCD. Data regarding this relationship are conflicting. We provide recommendations for improving the treatment of patients with SCD and depression. PMID- 11410539 TI - The overlap of depressive personality disorder and dysthymia, reconsidered. AB - This paper reviews empirical literature examining the diagnostic overlap of depressive personality disorder and dysthymia. Although some articles have suggested that depressive personality disorder lacks distinctiveness from dysthymia and may be one type of affective disorder, a large proportion of the literature may, in fact, support its distinctiveness as a personality disorder. Additional research is needed to determine the features of the two disorders that best distinguish between them, so that depressive personality disorder can be further evaluated for its validity and usefulness in the diagnostic nomenclature. PMID- 11410541 TI - "In sleep i almost never grope": blindness, neuropsychiatric deficits, and a chaotic upbringing. PMID- 11410542 TI - Ebb and flow: a theory of lasting relationships. PMID- 11410543 TI - The genomic approach to candidate genes. PMID- 11410544 TI - Do not resuscitate. PMID- 11410545 TI - Anomalous papillary muscle in hypertrophic cardiomyopathy. PMID- 11410546 TI - Antibiotics before dental procedures for endocarditis prophylaxis: back to the future. PMID- 11410547 TI - Stamps in cardiology: Aspirin. PMID- 11410548 TI - Living with a univentricular heart. PMID- 11410549 TI - Non-sustained ventricular tachycardia during carotid massage. PMID- 11410550 TI - Physical training as a therapeutic measure in chronic heart failure: time for recommendations. PMID- 11410551 TI - Self portrait of patient with severe cardiac pain awaiting surgery. PMID- 11410552 TI - Role of DNA testing for diagnosis, management, and genetic screening in long QT syndrome, hypertrophic cardiomyopathy, and Marfan syndrome. PMID- 11410553 TI - Electrical interference between two transvenous implantable defibrillator leads. PMID- 11410554 TI - Diagnostic exercise physiology in chronic heart failure. PMID- 11410555 TI - Left ventricular dysfunction, natriuretic peptides, and mortality in an urban population. AB - OBJECTIVE: To report the mortality of left ventricular systolic dysfunction (LVD), assessed objectively by echocardiography, and its association with natriuretic peptide hormones in a random sample of 1640 men and women aged 25-74 years from a geographical, urban population. METHODS: Left ventricular function was measured by echocardiography in 1640 attendees studied in 1992-3. LVD was defined as a left ventricular ejection fraction (LVEF) 30% (p < 0.001). The median (interquartile range) BNP concentration in those who died was 16.9 pg/ml (8.8-27) and 7.8 pg/ml (3.4-13) in survivors (p < 0.0001). Similarly, N-ANP had a median concentration of 2.35 ng/ml (1.32-3.36) in those with a fatal outcome and 1.27 ng/ml (0.9-2.0) in those alive at four years (p < 0.0001). Subjects with an LVEF /= 17.9 pg/ml compared with 6.8% if their BNP was below this concentration (p = 0.013). Multivariate analysis revealed the independent predictors of four year all cause mortality to be increasing age (p < 0.001), a BNP concentration >/= 17.9 pg/ml (p = 0.006), the presence of ischaemic heart disease (p = 0.03), and male sex (p = 0.04). CONCLUSIONS: LVD is associated with a considerable mortality rate in this population. BNP also independently predicts outcome. In addition to its role as a diagnostic aid in chronic heart failure and LVD, it provides prognostic information and clarifies the meaning of a given degree of LVD. PMID- 11410556 TI - Images in cardiology: Structural failure of a coronary stent. PMID- 11410557 TI - Increased brain and atrial natriuretic peptides in patients with chronic right ventricular pressure overload: correlation between plasma neurohormones and right ventricular dysfunction. AB - OBJECTIVE: To evaluate the role of plasma neurohormones in the diagnosis of asymptomatic or minimally symptomatic right ventricular dysfunction. SETTING: Tertiary cardiovascular referral centre. METHODS: Plasma brain natriuretic peptide (BNP) and atrial natriuretic peptide (ANP) concentrations were measured in 21 asymptomatic or minimally symptomatic patients with chronic right ventricular pressure overload caused by congenital heart disease, and in seven healthy volunteers. Right ventricular ejection fraction was determined using magnetic resonance imaging. RESULTS: Right ventricular ejection fraction in the volunteers was higher than in the patients (69.0 (8.2)% v 58.0(12.0)%, respectively; p < 0.006). Left ventricular ejection fraction was 72.3(7.8)% in volunteers and 68.1(11.0)% in patients (NS). There was a significant difference between patients and volunteers in the plasma concentrations of BNP (5.3 (3.5) v 2.3 (1.7) pmol/l, respectively; p < 0.009) and ANP (7.3 (4.5) v 3.6 (1.4) pmol/l; p < 0.05). In both patients and volunteers, mean plasma ANP was higher than mean plasma BNP. Right ventricular ejection fraction was inversely correlated with BNP and ANP (respectively, r = 0.65; p < 0.0002 and r = 0.61; p < 0.002). There was no correlation between left ventricular ejection fraction and BNP (r = 0.2; NS) or ANP (r = 0.52; NS). Similarly, no correlation was shown between the level of right ventricular systolic pressure and either plasma BNP (r = 0.20) or plasma ANP (r = 0.07). CONCLUSIONS: There was a significant inverse correlation between right ventricular ejection fraction and the plasma neurohormones BNP and ANP in asymptomatic or minimally symptomatic patients with right ventricular pressure overload and congenital heart disease. Monitoring changes in BNP and ANP may provide quantitative follow up of right ventricular dysfunction in these patients. PMID- 11410558 TI - Echocardiographic assessment of arrhythmogenic right ventricular cardiomyopathy. AB - OBJECTIVE: To evaluate new echocardiographic modes in the diagnosis of arrhythmogenic right ventricular cardiomyopathy (ARVC). DESIGN: Prospective observational study. SETTING: University Hospital. SUBJECTS: 15 patients with ARVC and a control group of 25 healthy subjects. METHODS: Transthoracic echocardiography included cross sectional measurements of the right ventricular outflow tract, right ventricular inflow tract, and right ventricular body. Wall motion was analysed subjectively. M mode and pulsed tissue Doppler techniques were used for quantitative measurement of tricuspid annular motion at the lateral, septal, posterior, and anterior positions. Doppler assessment of tricuspid flow and systemic venous flow was also performed. RESULTS: Assessed by M mode, the total amplitude of the tricuspid annular motion was significantly decreased in the lateral, septal, and posterior positions in the patients compared with the controls. The tissue Doppler velocity pattern showed decreased early diastolic peak annular (E(A)) velocity and an accompanying decrease in early (E(A)) to late diastolic (A(A)) velocity ratio in all positions; the systolic annular velocity was significantly decreased only in the lateral position. Four patients had normal right ventricular dimensions and three were judged to have normal right ventricular wall motion. The patient group had also a significantly decreased tricuspid flow E:A ratio. CONCLUSIONS: Tricuspid annular measurements are valuable, easy to obtain, and allow quantitative assessment of right ventricular function. ARVC patients showed an abnormal velocity pattern that may be an early but non-specific sign of the disease. Normal right ventricular dimensions do not exclude ARVC, and subjective detection of early changes in wall motion may be difficult. PMID- 11410559 TI - Response of the QT interval to mental and physical stress in types LQT1 and LQT2 of the long QT syndrome. AB - OBJECTIVE: To study and compare the effects of mental and physical stress on long QT syndrome (LQTS) patients. DESIGN: Case-control study. MAIN OUTCOME MEASURES: QT intervals were measured from lead V3. Serum potassium and plasma catecholamine concentrations were also monitored. PATIENTS: 16 patients with type 1 LQTS (LQT1), 14 with type 2 LQTS (LQT2), both groups asymptomatic, and 14 healthy control subjects. INTERVENTIONS: Three types of mental stress tests and a submaximal exercise stress test. RESULTS: Heart rate responses to mental stress and exercise were similar in all groups. During mental stress, the mean QT interval shortened to a similar extent in controls (-29 ms), LQT1 patients (-34 ms), and LQT2 patients (-30 ms). During exercise, the corresponding QT adaptation to exercise stress was more pronounced (p < 0.01) in healthy controls (-47 ms) than in LQT1 (-38 ms) or LQT2 patients (-38 ms). During exercise changes in serum potassium concentrations were correlated to changes in QT intervals in controls, but not in LQTS patients. LQT1 and LQT2 patients did not differ in serum potassium, catecholamine or heart rate responses to mental or physical stress. CONCLUSIONS: QT adaptation to mental and exercise stress in healthy people and in patients with LQTS is different. In healthy people QT adaptation is more sensitive to physical than to mental stress while no such diverging pattern was seen in asymptomatic LQTS patients. PMID- 11410560 TI - Combined pacing and percutaneous closing device therapy for dilated cardiomyopathy and patent ductus arteriosus. PMID- 11410561 TI - Beta(2) adrenergic receptors mediate important electrophysiological effects in human ventricular myocardium. AB - OBJECTIVE: To define the effects of beta(2) adrenergic receptor stimulation on ventricular repolarisation in vivo. DESIGN: Prospective study. SETTING: Tertiary referral centre. PATIENTS: 85 patients with coronary artery disease and 22 normal controls. INTERVENTIONS: Intravenous and intracoronary salbutamol (a beta(2) adrenergic receptor selective agonist; 10-30 microg/min and 1-10 microg/min), and intravenous isoprenaline (a mixed beta(1)/beta(2) adrenergic receptor agonist; 1 5 microg/min), infused during fixed atrial pacing. MAIN OUTCOME MEASURES: QT intervals, QT dispersion, monophasic action potential duration. RESULTS: In patients with coronary artery disease, salbutamol decreased QT(onset) and QT(peak) but increased QT(end) duration; QT(onset)-QT(peak) and QT(peak)-QT(end) intervals increased, resulting in T wave prolongation (mean (SEM): 201 (2) ms to 233 (2) ms; p < 0.01). There was a large increase in dispersion of QT(onset), QT(peak), and QT(end) which was more pronounced in patients with coronary artery disease-for example, QT(end) dispersion: 50 (2) ms baseline v 98 (4) ms salbutamol (controls), and 70 (1) ms baseline v 108 (3) ms salbutamol (coronary artery disease); p < 0.001. Similar responses were obtained with isoprenaline. Monophasic action potential duration at 90% repolarisation shortened during intracoronary infusion of salbutamol, from 278 (4.1) ms to 257 (3.8) ms (p < 0.05). CONCLUSIONS: beta(2) adrenergic receptors mediate important electrophysiological effects in human ventricular myocardium. The increase in dispersion of repolarisation provides a mechanism whereby catecholamines acting through this receptor subtype may trigger ventricular arrhythmias. PMID- 11410562 TI - Long axis excursion in aortic stenosis. AB - OBJECTIVES: To examine long axis excursion in patients with all grades of aortic stenosis and preserved transverse systolic function, and to compare long axis excursion in symptomatic with that in asymptomatic severe aortic stenosis. DESIGN: Prospective comparative study. SETTING: Regional cardiothoracic centre. PATIENTS: 78 patients with all grades of aortic stenosis and normal fractional shortening and ejection fraction were studied. There were two comparison groups, 10 age matched normal subjects and 14 patients with aortic stenosis and fractional shortening < 26%. METHODS: Aortic valve function and left ventricular mass were assessed echocardiographically. M mode measurements of long axis excursion at the septal and lateral sides of the mitral annulus were taken. RESULTS: There were significant differences between the groups in long axis excursion at both the septal (p < 0.0001) and lateral sides of the mitral annulus (p = 0.002 by analysis of variance). Long axis excursion was independently related to both left ventricular mass index (p = 0.001) and the grade of aortic stenosis (p = 0.002). Comparing patients with severe aortic stenosis with and without symptoms, there were significant differences in effective orifice area (p = 0.02 ) and long axis excursion at the lateral side of the mitral annulus (p = 0.04), but not in fractional shortening, ejection fraction, or peak or mean pressure difference. CONCLUSION: In patients with aortic stenosis, long axis excursion is reduced even in the presence of normal fractional shortening or ejection fraction. It is lower in patients with symptomatic compared with asymptomatic severe aortic stenosis and may be of use in predicting the onset of symptoms. PMID- 11410563 TI - Outlet strut fracture of Bjork-Shiley convexo concave heart valves: the UK cohort study. AB - OBJECTIVE: To investigate the risk of outlet strut fracture (OSF) in Bjork-Shiley convexo concave (BSCC) valves in relation to patients' clinical and valve characteristics. DESIGN: A cohort of 2977 patients with 3325 valves with a follow up of 18 years. SETTING: 38 cardiac implantation centres in the UK. RESULTS: 56 OSF events were reported with 43 occurring in mitral and 13 in aortic valves. The overall OSF rate was 0.17%/year. No dominant clinical factor of risk was found, but multiple regression analysis identified age, body surface area, valve size, shop order fracture rate, and manufacturing period as risk factors for OSF. A 4% (95% confidence interval (CI) 2% to 6%) decrease in the risk of OSF was observed for each advancing year of age and a fivefold (95% CI 2 to 13) increase in risk for a 0.5 m(2) increase in body surface area. The association between the risk of OSF and valve size was not constant over time. Excess risks among 31 mm and 33 mm sizes (mainly mitral valves) decreased over time while that for 23 mm (almost all aortic valves) increased. The risk of OSF increased by 40% (95% CI 20% to 50%) for a unit increase in the fracture rate of other valves in the same batch. For valves manufactured during 1981 to 1984 the risk of OSF was 4 (95% CI 2 to 12) times greater than for valves manufactured before 1981. CONCLUSIONS: The OSF rates for 60 degrees BSCC valves observed in the UK are the highest among all monitored populations. The changing patterns of mitral and aortic valve OSF rates over time observed in this study have not been identified previously and highlight the need for continued monitoring of patients with the BSCC valve. PMID- 11410564 TI - Long term results of mechanical prostheses for treatment of active infective endocarditis. AB - OBJECTIVE: To analyse the long term results of mechanical prostheses for treating active infective endocarditis. DESIGN: Prospective cohort study of a consecutive series of patients diagnosed with infective endocarditis and operated on in the active phase of the infection for insertion of a mechanical prosthesis. SETTING: Tertiary referral centre in a metropolitan area. RESULTS: Between 1975 and 1997, 637 cases of infective endocarditis were diagnosed in the centre. Of these, 436 were left sided (with overall mortality of 20.3%). Surgical treatment in the active phase of the infection was needed in 141 patients (72% native, 28% prosthetic infective endocarditis). Mechanical prostheses were used in 131 patients. Operative mortality was 30.5% (40 patients). Ninety one survivors were followed up prospectively for (mean (SD)) 5.4 (4.5) years. Thirteen patients developed prosthetic valve dysfunction. Nine patients suffered reinfection: four of these (4%) were early and five were late. The median time from surgery for late reinfection was 1.4 years. During follow up, 12 patients died. Excluding operative mortality, actuarial survival was 86.6% at five years and 83.7% at 10 years; actuarial survival free from death, reoperation, and reinfection was 73.1% at five years and 59.8% at 10 years. CONCLUSIONS: In patients surviving acute infective endocarditis and receiving mechanical prostheses, the rate of early reinfection compares well with reported results of homografts. In addition, prosthesis dysfunction rate is low and long term survival is good. These data should prove useful for comparison with long term studies, when available, using other types of valve surgery in active infective endocarditis. PMID- 11410565 TI - Quality of life and perceived health status in surviving adults with univentricular heart. AB - OBJECTIVE: To evaluate the quality of life in patients with univentricular heart and to determine the impact of sociodemographic and clinical characteristics. DESIGN AND SETTING: Retrospective, cross sectional study conducted in a regional paediatric cardiology centre. PATIENTS: The health records of 89 survivors with univentricular heart (median age 21 years; range 17-49 years) were reviewed. Sixty seven answered the Duke questionnaire. Sociodemographic and clinical variables were similar in the responders and non-responders. The impact of sociodemographic and clinical variables on individual Duke's measures was assessed. RESULTS: The Duke scores of adults with univentricular heart were similar to the normal population. Cyanosis predicted a worse score for physical (p = 0.05) and perceived health measures (p = 0.02). A higher educational level predicted a better score for physical (p = 0.004), mental (p = 0.01), and general health measures (p = 0.02). Orthopaedic problems worsened the social score (p = 0.05). Psychosocial problems worsened the pain score (p = 0.04). In comparison with the other anatomical types, mitral atresia worsened the perceived health score (p = 0.02). Patients younger than 23 years scored better for almost all health and dysfunction measures. CONCLUSIONS: Despite repeated interventions and other disease related everyday stresses, a selected group of adults with univentricular heart had a satisfying quality of life. PMID- 11410566 TI - Endomyocardial fibrosis mimicking a dilated cardiomyopathy in a child. PMID- 11410567 TI - What do adult patients with congenital heart disease know about their disease, treatment, and prevention of complications? A call for structured patient education. AB - OBJECTIVE: To assess how much adults with congenital heart disease understand about their heart defect, its treatment, and the preventive measures necessary to avoid complications. DESIGN: Descriptive, cross sectional study. SETTING: Adult congenital heart disease programme in one tertiary care centre in Belgium. PATIENTS: 62 adults with congenital heart disease (47 men; 15 women), median age 23 years. MAIN OUTCOME MEASURES: Patients' knowledge was assessed during an outpatient visit using the Leuven knowledge questionnaire for congenital heart diseases, a 33 item instrument developed for this study. RESULTS: Patients had adequate knowledge (> 80% correct answers) about their treatment, frequency of follow up, dental practices, occupational choices, appropriateness of oral contraceptives, and the risks of pregnancy. Knowledge about the name and anatomy of the heart defect, the possibility of recurrent episodes of endocarditis during their lifetime, and the appropriateness of different physical activities was moderate (50-80% correct answers). There was poor understanding (< 50% correct answers) about the reasons for follow up, the symptoms of deterioration of the heart disease, the definition, characteristics, and risk factors of endocarditis, the impact of smoking and alcohol on the heart disease, the hereditary nature of the condition, and the suitability of intrauterine devices as contraceptives. CONCLUSIONS: Adults with congenital heart disease have important gaps in their knowledge about their condition. The results of this study can be used as a basis for developing or optimising structured educational interventions to enhance patients' health behaviour. PMID- 11410568 TI - Superior vena cava syndrome caused by multiple pacing leads. PMID- 11410569 TI - Effect of levosimendan on myocardial contractility, coronary and peripheral blood flow, and arrhythmias during coronary artery ligation and reperfusion in the in vivo pig model. AB - OBJECTIVE: To determine whether levosimendan, a calcium sensitiser that facilitates the activation of the contractile apparatus by calcium, improves myocardial contractile function during severe ischaemia and reperfusion without exacerbating the incidence of arrhythmias. DESIGN: Pigs were pretreated orally twice daily for 10 days with 0.08 mg/kg levosimendan or placebo. On day 11 the left main coronary artery was ligated for 30 minutes, followed by 30 minutes of reperfusion. A bolus dose of levosimendan, 11.2 microg/kg intravenously, or placebo was given 30 minutes before coronary ligation, followed by a continuous infusion of 0.2 microg/kg/min levosimendan or placebo for the remainder of the experiment. RESULTS: During the ischaemic period, cardiac output was higher in the levosimendan group than in the placebo group (mean (SD): 2.6 (0.5) v 2.0 (0.2) l/min, p < 0.05) and systemic vascular resistance was lower (2024 (188) v 2669 (424) dyne.s(-1).cm(-5), p < 0.005). During reperfusion, cardiac output and contractility (LV(max)dP/dt (pos), 956 (118) v 784 (130) mm Hg/s, p < 0.05) were increased by levosimendan. The incidence of ischaemic ventricular fibrillation and tachycardia was similar in the two groups but there were more arrhythmic events (ventricular tachycardia and ventricular fibrillation) in the levosimendan treated group (8/12 levosimendan v 1/9 control p = 0.05). CONCLUSIONS: Levosimendan improved cardiac output and myocardial contractility during coronary artery ligation and reperfusion. However, it increased the number of arrhythmic events during ischaemia in this model of in vivo regional ischaemia. PMID- 11410570 TI - Severe tricuspid regurgitation 14 years after diagnosis of "transient neonatal tricuspid regurgitation". AB - A 14 year old girl presented with severe tricuspid regurgitation after she was diagnosed with "transient tricuspid regurgitation of the newborn". In the neonatal period she had presented with severe tricuspid regurgitation without an obvious underlying anatomical cause. This spontaneously regressed during the first months of life. She was dismissed from follow up at the age of 5 years after complete normalisation of the clinical and echocardiographic examination. The subsequent evolution and management of the patient, as well as the possible pathogenesis responsible for the unusual clinical course, is discussed. This case stresses the importance of long term follow up of patients with transient tricuspid regurgitation. PMID- 11410571 TI - "Kissing abscess" of the anterior mitral valve leaflet from a vegetation on the non-coronary aortic cusp. PMID- 11410572 TI - Symptom expectations and delay in acute myocardial infarction patients. PMID- 11410573 TI - Elevated concentrations of macrophage colony stimulating factor predict worse in hospital prognosis in unstable angina. PMID- 11410574 TI - Use of cardiac rehabilitation among patients following coronary artery bypass surgery. PMID- 11410575 TI - Angiotensin receptor blockers for chronic heart failure and acute myocardial infarction. PMID- 11410576 TI - Role of stenting in coronary revascularisation. PMID- 11410577 TI - Matching the right drug to the right patient in essential hypertension. PMID- 11410578 TI - Rapid regression of primary pulmonary hypertension. AB - A 14 month old child presented for investigation of tachypnoea. No parenchymal lung disease was shown on chest x ray. On echocardiography there was normal intracardiac anatomy with significant pulmonary hypertension. At cardiac catheterisation the presence of primary pulmonary hypertension was confirmed, with a partial response to inhaled nitric oxide (80 ppm) and 100% oxygen. The child was referred for assessment for heart-lung transplantation while maintained on oxygen, inhaled nitric oxide, and nifedipine. Repeat cardiac catheterisation two months after presentation showed complete normalisation of the pulmonary artery pressures. PMID- 11410579 TI - Leiomyosarcoma of the right ventricle extending into the pulmonary trunk. AB - Primary leiomyosarcomas of the heart, particularly those affecting the right ventricle, are uncommon. A 70 year old woman presenting with the symptoms of progressive exertional dyspnoea and left sided pleuritic pain is presented. A leiomyosarcoma was diagnosed that originated in the right lateral ventricle wall, causing pulmonary outflow obstruction. Pathological analysis showed a neoplasm with a myxoid stroma, high mitotic activity, and nuclei expressing atypia. Immunohistochemical staining was found positive for vimentin and desmin. Seven months after complete surgical resection the tumour relapsed. This case illustrates the poor outcome, high relapse rate, and inefficiency of treatment associated with primary cardiac leiomyosarcomas. The current literature regarding incidence, diagnostic techniques, treatment strategies, and survival rates of this rare but terminal disease is reviewed. PMID- 11410580 TI - Catheter ablation of a monofocal premature ventricular complex triggering idiopathic ventricular fibrillation. AB - A 62 year old man was admitted for evaluation of recurrent episodes of syncope. A surface ECG showed frequent repetitive premature ventricular complexes of right ventricular outflow tract origin. Ventricular fibrillation was inducible by programmed electrical stimulation but otherwise cardiac evaluation was unremarkable. A diagnosis of idiopathic ventricular fibrillation was made and an implantable cardioverter-defibrillator (ICD) was installed. However, spontaneous ventricular fibrillation recurred, requiring repeated ICD discharges. The ventricular fibrillation was reproducibly triggered by a single premature ventricular complex with a specific QRS morphology. Radiofrequency catheter ablation was carried out to eradicate this complex. No ventricular fibrillation has developed after this procedure, and the patient does not require drug treatment. PMID- 11410581 TI - Effects of carvedilol on left ventricular function, mass, and scintigraphic findings in isolated left ventricular non-compaction. AB - A four month old infant with isolated left ventricular non-compaction was treated with carvedilol. Haemodynamic studies and various types of imaging-including echocardiography, radiographic angiography, magnetic resonance imaging, and single photon emission computed tomography with (201)Tl, (123)I-beta methyliodophenylpentadecanoic acid (BMIPP), and (123)I-metaiodobenzylguanidine (MIBG)-were performed before and 14 months after treatment. Left ventricular ejection fraction increased from 30% to 57%, and left ventricular end diastolic volume, end systolic volume, and end diastolic pressure showed striking reductions during treatment. Left ventricular mass decreased to about two thirds of the baseline value after treatment. Per cent wall thickening increased after carvedilol in the segments corresponding to non-compacted myocardium. A mismatch between (201)Tl and BMIPP uptake in the area of non-compaction observed before carvedilol disappeared after treatment. Impaired sympathetic neuronal function shown by MIBG recovered after treatment. Thus carvedilol had beneficial effects on left ventricular function, hypertrophy, and both metabolic and adrenergic abnormalities in isolated left ventricular non-compaction. PMID- 11410582 TI - Persistence of the eustachian valve in secundum atrial septal defects: possible implications for cerebral embolism and transcatheter closure procedures. AB - Transcatheter closure of large secundum atrial septal defects is now accepted clinical practice. With the introduction of easily applicable closure devices the indications for this procedure have been expanded to include the closure of patent foramen ovale after cerebral stroke of unknown origin. In some of these patients a persistent eustachian valve is present. The clinical relevance of this finding is still unclear. A 36 year old patient with a brainstem stroke of unknown origin and a secundum atrial septal defect in combination with a persisting prominent eustachian valve is reported. The potential role of the eustachian valve in the genesis of the stroke and the difficulties during transcatheter closure of the defect because of the persisting valve are discussed. PMID- 11410583 TI - Unusual localisation of a ventricular septal defect following blunt chest trauma. AB - A 64 year old man presented with a traumatic ventricular septal defect following blunt chest trauma 40 years before. Echocardiography and left ventriculography were helpful in locating the unusual septal defect, which was subpulmonary. The shunt was small, but the anomalous chronic overload led to right ventricular failure. The surgical correction was thus too late to improve right ventricular function. PMID- 11410584 TI - A distinct meta-active conformation in the 1.1-A resolution structure of wild type ApoCheY. AB - CheY is the best characterized member of the response regulator superfamily, and as such it has become the principal model for understanding the initial molecular mechanisms of signaling in two-component systems. Normal signaling by response regulators requires phosphorylation, in combination with an activation mechanism whose conformational effects are not completely understood. CheY activation involves three events, phosphorylation, a conformational change in the beta(4)- alpha(4) loop, and a rotational restriction of the side chain of tyrosine 106. An outstanding question concerns the nature of an active conformation in the apoCheY population. The details of this 1.08-A resolution crystal structure of wild-type apoCheY shows the beta(4)--alpha(4) loop in two distinctly different conformations that sterically correlate with the two rotameric positions of the tyrosine 106 side chain. One of these conformational states of CheY is the inactive form, and we propose that the other is a meta-active form, responsible for the active properties seen in apoCheY. PMID- 11410585 TI - Epitope diversity of N-glycans from bovine peripheral myelin glycoprotein P0 revealed by mass spectrometry and nano probe magic angle spinning 1H NMR spectroscopy. AB - The carbohydrate structures present on the glycoproteins in the central and peripheral nerve systems are essential in many cell adhesion processes. The P0 glycoprotein, expressed by myelinating Schwann cells, plays an important role during the formation and maintenance of myelin, and it is the most abundant constituent of myelin. Using monoclonal antibodies, the homophilic binding of the P0 glycoprotein was shown to be mediated via the human natural keller cell (HNK) 1 epitope (3-O-SO(3)H-GlcUA(beta1-3)Gal(beta1-4)GlcNAc) present on the N-glycans. We recently described the structure of the N-glycan carrying the HNK-1 epitope, present on bovine peripheral myelin P0 (Voshol, H., van Zuylen, C. W. E. M., Orberger, G., Vliegenthart, J. F. G., and Schachner, M. (1996) J. Biol. Chem. 271, 22957-22960). In this study, we report on the structural characterization of the detectable glycoforms, present on the single N-glycosylation site, using state-of-the-art NMR and mass spectrometry techniques. Even though all structures belong to the hybrid- or biantennary complex-type structures, the variety of epitopes is remarkable. In addition to the 3-O-sulfate present on the HNK-1 carrying structures, most of the glycans contain a 6-O-sulfated N acetylglucosamine residue. This indicates the activity of a 6-O-sulfo-GlcNAc transferase, which has not been described before in peripheral nervous tissue. The presence of the disialo-, galactosyl-, and 6-O-sulfosialyl-Lewis X epitopes provides evidence for glycosyltransferase activities not detected until now. The finding of such an epitope diversity triggers questions related to their function and whether events, previously attributed merely to the HNK-1 epitope, could be mediated by the structures described here. PMID- 11410586 TI - Activation loop Ser744 and Ser748 in protein kinase D are transphosphorylated in vivo. AB - The importance of activation loop phosphorylation in the regulation of protein kinase D (PKD/protein kinase C (PKC) mu) activity has become controversial. In order to clarify the mechanism(s) of PKD activation, we developed a novel phosphospecific antibody recognizing phosphorylated Ser(748) in PKD (pS748). Western blot analysis with the pS748 antibody, carried out with a variety of PKD forms and in a variety of cell types including full-length PKD transfected in COS 7 and HEK 293 cells, a green fluorescent protein-PKD fusion protein transfected in either Swiss 3T3 fibroblasts or Madin-Darby canine kidney epithelial cells, and endogenous PKD expressed in A20 lymphocytes and Rat-1 fibroblasts, indicated that Ser(748) phosphorylation was absent from unstimulated cells. In contrast, dramatic increases in Ser(748) phosphorylation were induced by phorbol esters, bombesin, or cross-linking of B lymphocyte antigen receptors or by cotransfection with active PKCepsilon or PKCeta. Western analysis using a second phosphospecific antibody, which primarily recognizes PKD phosphorylated at Ser(744), revealed that Ser(744) phosphorylation accompanies Ser(748) phosphorylation during PKD activation in vivo. Ser(744)/Ser(748) phosphorylation requires PKC but not PKD activity, indicative of transphosphorylation. Our results provide new experimental evidence indicating that activation loop phosphorylation at Ser(744) and Ser(748) occurs during PKD activation in vivo and support the notion of a PKC PKD phosphorylation cascade. PMID- 11410587 TI - Rapid protein kinase D translocation in response to G protein-coupled receptor activation. Dependence on protein kinase C. AB - Protein kinase D (PKD)/protein kinase C (PKC) mu is a serine/threonine protein kinase that can be activated by physiological stimuli like growth factors, antigen-receptor engagement and G protein-coupled receptor (GPCR) agonists via a phosphorylation-dependent mechanism that requires PKC activity. In order to investigate the dynamic mechanisms associated with GPCR signaling, the intracellular translocation of a green fluorescent protein-tagged PKD was analyzed by real-time visualization in fibroblasts and epithelial cells stimulated with bombesin, a GPCR agonist. We found that bombesin induced a rapidly reversible plasma membrane translocation of green fluorescent protein tagged PKD, an event that can be divided into two distinct mechanistic steps. The first step, which is exclusively mediated by the cysteine-rich domain in the N terminus of PKD, involved its translocation from the cytosol to the plasma membrane. The second step, i.e. the rapid reverse translocation of PKD from the plasma membrane to the cytosol, required its catalytic domain and surprisingly PKC activity. These findings provide evidence for a novel mechanism by which PKC coordinates the translocation and activation of PKD in response to bombesin induced GPCR activation. PMID- 11410588 TI - Fibroblast quiescence in floating or released collagen matrices: contribution of the ERK signaling pathway and actin cytoskeletal organization. AB - Fibroblasts in attached collagen matrices proliferate, whereas cells in floating or released matrices become quiescent. Cells in attached matrices had prominent actin stress fibers, indicating that they were under isometric tension, whereas stress fibers were absent from fibroblasts in floating or released matrices. Compared with cells in attached matrices, cells in floating or released matrices showed down-regulation of cyclin D1 and up-regulation of p27(Kip1) cyclin dependent kinase inhibitor, and similar changes occurred after the ERK signaling pathway was blocked by UO126 in cells in attached matrices. A different pattern of changes in cell cycle regulatory proteins occurred, however, after serum deprivation or actin cytoskeletal depolymerization by latrunculin B, which did not prevent signaling through the ERK pathway. Therefore, cell quiescence in floating or released collagen matrices could be explained by decreased signaling through the ERK pathway, but these changes were not accounted for by the absence of isometric tension in the cells. PMID- 11410589 TI - ERKs regulate cyclic AMP-induced steroid synthesis through transcription of the steroidogenic acute regulatory (StAR) gene. AB - Cyclic AMP-dependent expression of the steroidogenic acute regulatory (StAR) protein is thought to be the controlling step for steroid production, but the mechanisms through which external signals are translated into increased transcription of the StAR gene are unknown. We demonstrate that cyclic AMP induced steroid synthesis is dependent upon the phosphorylation and activation of ERKs and that ERK activation results in enhanced phosphorylation of SF-1 and increased steroid production through increased transcription of the StAR gene. Adenylate cyclase activation with forskolin (FSK) caused a time-dependent increase in ERK activity and translocation from cytoplasm to nucleus, which correlated with an increase in StAR mRNA levels, StAR protein accumulation, and steroidogenesis. Similarly, ERK inhibition led to a reduction in the levels of FSK-stimulated StAR mRNA, StAR protein, and steroid secretion. These effects were attributed to the finding that ERK activity is required for SF-1 phosphorylation, a transcription factor required for the regulation of StAR gene transcription. This conclusion was supported by our demonstration of an ERK-dependent increase in the binding of SF-1 from FSK-treated Y1 nuclei to three consensus double stranded DNA sequences from the StAR promoter region. These observations suggest that the activation of ERK2/1 by increasing cAMP is an obligatory and regulated stage in the stimulation of steroid synthesis by cyclic AMP-generating stimuli. PMID- 11410590 TI - Serum- and glucocorticoid-inducible kinase SGK phosphorylates and negatively regulates B-Raf. AB - Phosphorylation can both positively and negatively regulate activity of the Raf kinases. Akt has been shown to phosphorylate and inhibit C-Raf activity. We have recently reported that Akt negatively regulates B-Raf kinase activation by phosphorylating multiple residues within its amino-terminal regulatory domain. Here we investigated the regulation of B-Raf by serum and glucocorticoid inducible kinase, SGK, which shares close sequence identity with the catalytic domain of Akt but lacks the pleckstrin homology domain. We observed that SGK inhibits B-Raf activity. A comparison of substrate specificity between SGK and Akt indicates that SGK is a potent negative regulator of B-Raf. In contrast to Akt, SGK negatively regulates B-Raf kinase activity by phosphorylating only a single Akt consensus site, Ser(364). Under similar experimental conditions, SGK displays a measurably stronger inhibitory effect on B-Raf kinase activity than Akt, whereas Akt exhibits a more inhibitory effect on the forkhead transcription factor, FKHR. The selective substrate specificity is correlated with an enhanced association between Akt or SGK and their preferred substrates, FKHR and B-Raf, respectively. These results indicate that B-Raf kinase activity is negatively regulated by Akt and SGK, suggesting that the cross-talk between the B-Raf and other signaling pathways can be mediated by both Akt and SGK. PMID- 11410591 TI - Complex formation and cooperation of protein kinase C theta and Akt1/protein kinase B alpha in the NF-kappa B transactivation cascade in Jurkat T cells. AB - Protein kinase C theta (PKC theta) is known to induce NF-kappa B, an essential transcriptional element in T cell receptor/CD28-mediated interleukin-2 production but also T cell survival. Here we provide evidence that PKC theta is physically and functionally coupled to Akt1 in this signaling pathway. First, T cell receptor/CD3 ligation was sufficient to induce activation as well as plasma membrane recruitment of PKC theta. Second, PKC theta selectively cooperated with Akt1, known to act downstream of CD28 co-receptor signaling, in activating a NF kappa B reporter in T cells. Third, Akt1 function was shown to be required for PKC theta-mediated NF-kappa B transactivation. Fourth, PKC theta co immunoprecipitated with Akt1; however, neither Akt1 nor PKC theta served as a prominent substrate for each other in vitro as well as in intact T cells. Finally, plasma membrane targeting of PKC theta and Akt1 exerted synergistic transactivation of the I-kappa B kinase beta/inhibitor of NF-kappa B/NF-kappa B signaling cascade independent of T cell activation. Taken together, these findings suggest a direct cross-talk between PKC theta and Akt1 in Jurkat T cells. PMID- 11410592 TI - Estrogen regulation of cyclin D1 gene expression in ZR-75 breast cancer cells involves multiple enhancer elements. AB - Cyclin D1 gene expression is induced by 17beta-estradiol (E2) in human breast cancer cells and is important for progression of cells through the G(1) phase of the cell cycle. The mechanism of activation of cyclin D1 is mitogen- and cell context-dependent, and this study describes the role of multiple promoter elements required for induction of cyclin D1 by E2 in estrogen receptor (ER) positive ZR-75 breast cancer cells. Transcriptional activation of cyclin D1 by E2 was dependent, in part, on a proximal cAMP-response element at -66, and this was linked to induction of protein kinase A-dependent pathways. These results contrasted to a recent report showing that induction of cyclin D1 by E2 in ER positive MCF-7 and HeLa cells was due to up-regulation of c-jun and subsequent interaction of c-Jun-ATF-2 with the CRE. Moreover, further examination of the proximal region of the cyclin D1 promoter showed that three GC-rich Sp1-binding sites at -143 to -110 were also E2-responsive, and interaction of ERalpha and Sp1 proteins at these sites was confirmed by electromobility shift and chromatin immunoprecipitation assays. Thus, induction of cyclin D1 by E2 in ZR-75 cells is regulated through nuclear ERalpha/Sp1 and epigenetic protein kinase A activation pathways, and our results suggest that this mechanism may be cell context dependent even among ER-positive breast cancer cell lines. PMID- 11410593 TI - The protein kinase YakA regulates g-protein-linked signaling responses during growth and development of Dictyostelium. AB - A genetic screen for Dictyostelium mutants that phenotypically resemble cells lacking the G-protein beta-subunit yielded the protein kinase YakA. Like gbeta null cells, yakA-null cells fail to enter development and display slow growth on bacterial lawns. We created a temperature-sensitive yakA mutant and showed that YakA activity is required not only at the onset but also during development. The yakA-null cells have strong defects in folic acid-induced responses, such as actin polymerization and cGMP accumulation, indicating that they play a role in G protein-mediated signaling responses. We propose that YakA acts downstream of G proteins, because cAMP receptors still couple to G-proteins in the yakA mutant. In addition, the previously observed growth arrest induced by overexpression of YakA also occurs in gbeta mutants. We localized YakA-GFP to the cytosol suggesting that YakA may be a functional homolog of its mammalian counterparts Dyrk2 and Dyrk3, a subclass of dual-specificity Yak-related kinases (Dyrk) with unknown function. PMID- 11410594 TI - Mitochondrial respiratory chain deficiency in Caenorhabditis elegans results in developmental arrest and increased life span. AB - The growth and development of Caenorhabditis elegans are energy-dependent and rely on the mitochondrial respiratory chain (MRC) as the major source of ATP. The MRC is composed of approximately 70 nuclear and 12 mitochondrial gene products. Complexes I and V are multisubunit proteins of the MRC. The nuo-1 gene encodes the NADH- and FMN-binding subunit of complex I, the NADH-ubiquinone oxidoreductase. The atp-2 gene encodes the active-site subunit of complex V, the ATP synthase. The nuo-1(ua1) and atp-2(ua2) mutations are both lethal. They result in developmental arrest at the third larval stage (L3), arrest of gonad development at the second larval stage (L2), and impaired mobility, pharyngeal pumping, and defecation. Surprisingly, the nuo-1 and atp-2 mutations significantly lengthen the life spans of the arrested animals. When MRC biogenesis is blocked by chloramphenicol or doxycycline (inhibitors of mitochondrial translation), a quantitative and homogeneous developmental arrest as L3 larvae also results. The common phenotype induced by the mutations and drugs suggests that the L3-to-L4 transition may involve an energy-sensing developmental checkpoint. Since approximately 200 gene products are needed for MRC assembly and mtDNA replication, transcription, and translation, we predict that L3 arrest will be characteristic of mutations in these genes. PMID- 11410595 TI - Atp11p and Atp12p are assembly factors for the F(1)-ATPase in human mitochondria. AB - Atp11p and Atp12p were first described as proteins required for assembly of the F(1) component of the mitochondrial ATP synthase in Saccharomyces cerevisiae (Ackerman, S. H., and Tzagoloff, A. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 4986-4990). Here we report the isolation of the cDNAs and the characterization of the human genes for Atp11p and Atp12p and show that the human proteins function like their yeast counterparts. Human ATP11 spans 24 kilobase pairs in 9 exons and maps to 1p32.3-p33, while ATP12 contains > or =8 exons and localizes to 17p11.2. Both genes are broadly conserved in eukaryotes and are expressed in a wide range of tissues, which suggests that Atp11p and Atp12p are essential housekeeping proteins of human cells. The information reported herein will be useful in the evaluation of patients with ascertained deficiencies in the ATP synthase, in which the underlying biochemical defect is unknown and may reside in a protein that influences the assembly of the enzyme. PMID- 11410596 TI - Water and ion permeation of aquaporin-1 in planar lipid bilayers. Major differences in structural determinants and stoichiometry. AB - The aquaporin-1 (AQP1) water channel protein is known to facilitate the rapid movement of water across cell membranes, but a proposed secondary role as an ion channel is still unsettled. Here we describe a method to simultaneously measure water permeability and ion conductance of purified human AQP1 after reconstitution into planar lipid bilayers. Water permeability was determined by measuring Na(+) concentrations adjacent to the membrane. Comparisons with the known single channel water permeability of AQP1 indicate that the planar lipid bilayers contain from 10(6) to 10(7) water channels. Addition of cGMP induced ion conductance in planar bilayers containing AQP1, whereas cAMP was without effect. The number of water channels exceeded the number of active ion channels by approximately 1 million-fold, yet p-chloromethylbenzenesulfonate inhibited the water permeability but not ion conductance. Identical ion channel parameters were achieved with AQP1 purified from human red blood cells or AQP1 heterologously expressed in Saccharomyces cerevisae and affinity purified with either N- or C terminal poly-histidine tags. Rp-8-Br-cGMP inhibited all of the observed conductance levels of the cation selective channel (2, 6, and 10 pS in 100 mm Na(+) or K(+)). Deletion of the putative cGMP binding motif at the C terminus by introduction of a stop codon at position 237 yielded a truncated AQP1 protein that was still permeated by water but not by ions. Our studies demonstrate a method for simultaneously measuring water permeability and ion conductance of AQP1 reconstituted into planar lipid bilayers. The ion conductance occurs (i) through a pathway distinct from the aqueous pathway, (ii) when stimulated directly by cGMP, and (iii) in only an exceedingly small fraction of AQP1 molecules. PMID- 11410597 TI - Inherited Brugada and long QT-3 syndrome mutations of a single residue of the cardiac sodium channel confer distinct channel and clinical phenotypes. AB - Defects of the SCN5A gene encoding the cardiac sodium channel alpha-subunit are associated with both the long QT-3 (LQT-3) subtype of long-QT syndrome and Brugada syndrome (BrS). One previously described SCN5A mutation (1795insD) in the C terminus results in a clinical phenotype combining QT prolongation and ST segment elevation, indicating a close interrelationship between the two disorders. Here we provide additional evidence that these two disorders are closely related. We report the analysis of two novel mutations on the same codon, Y1795C (LQT-3) and Y1795H (BrS), expressed in HEK 293 cells and characterized using whole-cell patch clamp procedures. We find marked and opposing effects on channel gating consistent with activity associated with the cellular basis of each clinical disorder. Y1795H speeds and Y1795C slows the onset of inactivation. The Y1795H, but not the Y1795C, mutation causes a marked negative shift in the voltage dependence of inactivation, and neither mutation affects the kinetics of the recovery from inactivation. Interestingly, both mutations increase the expression of sustained Na+ channel activity compared with wild type (WT) channels, although this effect is most pronounced for the Y1795C mutation, and both mutations promote entrance into an intermediate or a slowly developing inactivated state. These data confirm the key role of the C-terminal tail of the cardiac Na+ channel in the control of channel gating, illustrate how subtle changes in channel biophysics can have significant and distinct effects in human disease, and, additionally, provide further evidence of the close interrelationship between BrS and LQT-3 at the molecular level. PMID- 11410598 TI - Site-specific protease activity of the carboxyl-terminal domain of Semliki Forest virus replicase protein nsP2. AB - The virus-specific components (nsP1-nsP4) of Semliki Forest virus RNA polymerase are synthesized as a large polyprotein (P1234), which is cleaved by a virus encoded protease. Based on mutagenesis studies, nsP2 has been implicated as the protease moiety of P1234. Here, we show that purified nsP2 (799 amino acids) and its C-terminal domain Pro39 (amino acids 459-799) specifically process P1234 and its cleavage intermediates. Analysis of cleavage products of in vitro synthesized P12, P23, and P34 revealed cleavages at sites 1/2, 2/3, and 3/4. The cleavage regions of P1/2, P2/3, and P3/4 were expressed as thioredoxin fusion proteins (Trx12, Trx23, and Trx34), containing approximately 20 amino acids on each side of the cleavage sites. After exposure of these purified fusion proteins to nsP2 or Pro39, the reaction products were analyzed by SDS-polyacrylamide gel electrophoresis, mass spectrometry, and amino-terminal sequencing. The expected amino termini of nsP2, nsP3, and nsP4 were detected. The cleavage at 3/4 site was most efficient, whereas cleavage at 1/2 site required 5000-fold more of Pro39, and 2/3 site was almost resistant to cleavage. The activity of Pro39 was inhibited by N-ethylmaleimide, Zn(2+), and Cu(2+), but not by EDTA, phenylmethylsulfonyl fluoride, or pepstatin, in accordance with the thiol proteinase nature of nsP2. PMID- 11410599 TI - Nuclear factor kappa B is a molecular target for sulforaphane-mediated anti inflammatory mechanisms. AB - Sulforaphane (SFN), an aliphatic isothiocyanate, is a known cancer chemopreventive agent. Aiming to investigate anti-inflammatory mechanisms of SFN, we here report a potent decrease in lipopolysaccharide (LPS)-induced secretion of pro-inflammatory and pro-carcinogenic signaling factors in cultured Raw 264.7 macrophages after SFN treatment, i.e. NO, prostaglandin E(2), and tumor necrosis factor alpha. SFN did not directly interact with NO, nor did it inhibit inducible nitric-oxide synthase enzymatic activity. Western blot analyses revealed time- and dose-dependent reduction of LPS-induced inducible nitric-oxide synthase as well as Cox-2 protein expression, which was suppressed at the transcriptional level. To reveal the target of SFN beyond its anti-inflammatory action, we performed electrophoretic mobility shift assay analyses of transcription factor DNA binding. Consequently, nuclear factor kappa B (NF-kappa B), a pivotal transcription factor in LPS-stimulated pro-inflammatory response, was identified as the key mediator. SFN selectively reduced DNA binding of NF-kappa B without interfering with LPS-induced degradation of the inhibitor of NF-kappa B nor with nuclear translocation of NF-kappa B. Because SFN can interact with thiol groups by dithiocarbamate formation, it may impair the redox-sensitive DNA binding and transactivation of NF-kappa B. Sulforaphane could either directly inactivate NF kappa B subunits by binding to essential Cys residues or interact with glutathione or other redox regulators like thioredoxin and Ref-1 relevant for NF kappa B function. Our data provide novel evidence that anti-inflammatory mechanisms contribute to sulforaphane-mediated cancer chemoprevention. PMID- 11410600 TI - S100A13 participates in the release of fibroblast growth factor 1 in response to heat shock in vitro. AB - S100A13, a member of the S100 gene family of Ca(2+)-binding proteins has been previously characterized as a component of a brain-derived heparin-binding multiprotein aggregate/complex containing fibroblast growth factor 1 (FGF1). We report that while expression of S100A13 in NIH 3T3 cells results in the constitutive release of S100A13 into the extracellular compartment at 37 degrees C, co-expression of S100A13 with FGF1 represses the constitutive release of S100A13 and enables NIH 3T3 cells to release S100A13 in response to temperature stress. S100A13 release in response to stress occurs with kinetics similar to that observed for the stress-induced release of FGF1, but S100A13 expression is able to reverse the sensitivity of FGF1 release to inhibitors of transcription and translation. The release of FGF1 and S100A13 in response to heat shock results in the solubility of FGF1 at 100% (w/v) ammonium sulfate saturation, and the expression of a S100A13 deletion mutant lacking its novel basic residue-rich domain acts as a dominant negative effector of FGF1 release in vitro. Surprisingly, the expression of S100A13 also results in the stress-induced release of a Cys-free FGF1 mutant, which is normally not released from NIH 3T3 cells in response to heat shock. These data suggest that S100A13 may be a component of the pathway for the release of the signal peptide-less polypeptide, FGF1, and may involve a role for S100A13 in the formation of a noncovalent FGF1 homodimer. PMID- 11410601 TI - Is Congo red an amyloid-specific dye? AB - Congo red (CR) binding, monitored by characteristic yellow-green birefringence under crossed polarization has been used as a diagnostic test for the presence of amyloid in tissue sections for several decades. This assay is also widely used for the characterization of in vitro amyloid fibrils. In order to probe the structural specificity of Congo red binding to amyloid fibrils we have used an induced circular dichroism (CD) assay. Amyloid fibrils from insulin and the variable domain of Ig light chain demonstrate induced CD spectra upon binding to Congo red. Surprisingly, the native conformations of insulin and Ig light chain also induced Congo red circular dichroism, but with different spectral shapes than those from fibrils. In fact, a wide variety of native proteins exhibited induced CR circular dichroism indicating that CR bound to representative proteins from different classes of secondary structure such as alpha (citrate synthase), alpha + beta (lysozyme), beta (concavalin A), and parallel beta-helical proteins (pectate lyase). Partially folded intermediates of apomyoglobin induced different Congo red CD bands than the corresponding native conformation, however, no induced CD bands were observed with unfolded protein. Congo red was also found to induce oligomerization of native proteins, as demonstrated by covalent cross linking and small angle x-ray scattering. Our data suggest that Congo red is sandwiched between two protein molecules causing protein oligomerization. The fact that Congo red binds to native, partially folded conformations and amyloid fibrils of several proteins shows that it must be used with caution as a diagnostic test for the presence of amyloid fibrils in vitro. PMID- 11410605 TI - Correlative fluorescence and electron microscopy on ultrathin cryosections: bridging the resolution gap. AB - Microscopy has become increasingly important for analysis of cells and cell function in recent years. This is due in large part to advances in light microscopy that facilitate quantitative studies and improve imaging of living cells. Analysis of fluorescence signals has often been a key feature in these advances. Such studies involve a number of techniques, including imaging of fluorescently labeled proteins in living cells, single-cell physiological experiments using fluorescent indicator probes, and immunofluorescence localization. The importance of fluorescence microscopy notwithstanding, there are instances in which electron microscopy provides unique information about cell structure and function. Correlative microscopy in which a fluorescence signal is reconciled with a signal from the electron microscope is an additional tool that can provide powerful information for cellular analysis. Here we review two different methodologies for correlative fluorescence and electron microscopy using ultrathin cryosections and the advantages attendant on this approach. (J Histochem Cytochem 49:803-808, 2001) PMID- 11410606 TI - High-resolution CryoFESEM of individual cell adhesion molecules (CAMs) in the glycocalyx of human platelets: detection of P-selectin (CD62P), GPI-IX complex (CD42A/CD42B alpha,B beta), and integrin GPIIbIIIa (CD41/CD61) by immunogold labeling and stereo imaging. AB - The aim of this study was to develop a model for the detection of individual cell adhesion molecules (CAMs) in the glycocalyx of spread human platelets using high resolution cryo-field emission scanning electron microscopy (cryoFESEM). Three surface glycoprotein CAMs, P-selectin (CD62P), GPIba in the GPI-IX complex (CD42a/CD42b alpha,b beta), and the integrin GPIIbIIIa (CD41/CD61) in the human platelet were selected on the basis of their unique topographic shape. Spread human platelets were indirectly immunolabeled with 10-nm colloidal gold and then cryoimmobilized. After sublimation of water from the cryoimmobilized sample, partially freeze-dried platelets were coated unidirectionally with Pt, stabilized with carbon, and examined in an in-lens cryoFESEM using high-resolution backscattered electron imaging. CAMs were detected by indirect immunogold labeling and the length of each type of CAM was determined using analysis of differences in parallax as measured in the software program Sterecon. Our results demonstrate the efficacy of using high-resolution cryoFESEM to recognize and detect individual CAMs in the glycocalyx. Further advances in production of metal coatings with finer granularity, together with improvements in imaging (tilting and angle of stereo images), may provide better definition of the topography associated with glycosylation and formation of multimeric CAM complexes. (J Histochem Cytochem 49:809-819, 2001) PMID- 11410607 TI - Advances in cytochemical methods for detection of apoptosis. AB - In an earlier article from this laboratory, the current methods developed to detect apoptosis in cells and tissues were highlighted, along with the challenges in their interpretation. Recent discoveries concerning the underlying biochemical mechanisms of apoptotic effector pathways have made possible further assays that allow a more direct measure of the activation of the apoptotic machinery in cells. This article summarizes some of these newer methods and extends the interpretation of the more classical assays of apoptosis in a defined cell system. We present data in KB and PC3 cell model culture systems induced to undergo apoptosis by the plant toxin ricin. Using a modified in situ nick translation assay (ISNT) with either Bodipy or BUdR labeling, we confirm that most cells showing altered nuclear morphology do not show reactivity with this assay until very late in the apoptotic process. We also show that only a minority of cells label with fluorescent annexin V during apoptosis but that apoptotic cells continue to internalize material from the cell surface through endocytosis after becoming reactive with annexin V. In addition, we describe the utility of a prototype of new assays for caspase substrate cleavage products, the detection of cleaved cytokeratin 18. It is these newer cleavage product assays that perhaps hold the greatest promise for specific detection of apoptosis in cells either in cell culture or in intact tissues. (J Histochem Cytochem 49:821-832, 2001) PMID- 11410608 TI - Kalirin, a GDP/GTP exchange factor of the Dbl family, is localized to nerve, muscle, and endocrine tissue during embryonic rat development. AB - Kalirin, a homologue of trio and UNC-73, has been previously demonstrated to cause cytoskeletal rearrangements, enhanced outgrowth of neuritic processes, and altered secretion. In the adult rat, kalirin is specifically localized to the central nervous system, with the main adult isoform, kalirin-7, concentrated in neuronal postsynaptic densities. In this study we examined the expression of kalirin in rat tissue from embryonic Day 10 (E10) through E18, using an antibody that detects all known kalirin isoforms. Kalirin expression in the embryo was more widespread than in the adult, with localization of kalirin protein to both neuronal and non-neuronal tissue, such as muscle, lung, intestinal epithelium, and pancreas. In neurons, kalirin was localized both in cell bodies and axon processes; in muscle tissue, kalirin was highly localized to migrating myogenic cells and at muscle attachment sites. Western blotting analysis indicated that kalirin-7, the major adult isoform, was a minor component of embryonic kalirin; the main isoform expressed in the embryo was kalirin-9. This is the first identification of kalirin expression in embryonic tissue and the first demonstration of non-neuronal expression of kalirin. (J Histochem Cytochem 49:833 844, 2001) PMID- 11410609 TI - Cell type-specific localization of sphingosine kinase 1a in human tissues. AB - Cell type-specific localization of sphingosine kinase 1a (SPHK1a) in tissues was analyzed with a rabbit polyclonal antibody against the 16 C-terminal amino acids derived from the recently reported mouse cDNA sequence of SPHK1a. This antibody (anti-SPHK1a antibody) can react specifically with SPHK1a of mouse, rat, and human tissues. Utilizing its crossreactivity to human SPHK1a, the cell-specific localization of SPHK1a in human tissues was histochemically examined. Strong positive staining for SPHK1a was observed in the white matter in the cerebrum and cerebellum, the red nucleus and cerebral peduncle in the midbrain, the uriniferous tubules in the kidney, the endothelial cells in vessels of various organs, and in megakaryocytes and platelets. The lining cells of sinusoids in the liver and splenic cords in the spleen showed moderate staining. Columnar epithelia in the intestine and Leydig's cells in the testis showed weak staining patterns. In addition, TPA-treated HEL cells, a human leukemia cell line, showed a megakaryocytic phenotype accompanied with increases in immunostaining of both SPHK1a and SPHK enzyme activity, suggesting that SPHK1a may be a novel marker of megakaryocytic differentiation and that this antibody is also useful for in vitro study of differentiation models.(J Histochem Cytochem 49:845-855, 2001) PMID- 11410610 TI - Variations in dystrophin complex in red and white caudal muscles from Torpedo marmorata. AB - We present an up-to-date study on the nature, at the protein level, of various members of the dystrophin complex at the muscle cell membrane by comparing red and white caudal muscles from Torpedo marmorata. Our investigations involved immunodetection approaches and Western blotting analysis. We determined the presence or absence of different molecules belonging to the dystrophin family complex by analyzing their localization and molecular weight. Specific antibodies directed against dystrophin, i.e., DRP2 alpha-dystrobrevin, beta-dystroglycan, alpha-syntrophin, alpha-, beta-, gamma-, and delta-sarcoglycan, and sarcospan, were used. The immunofluorescence study (confocal microscopy) showed differences in positive immunoreactions at the sarcolemmal membrane in these slow-type and fast-type skeletal muscle fibers. Protein extracts from T. marmorata red and white muscles were analyzed by Western blotting and confirmed the presence of dystrophin and associated proteins at the expected molecular weights. Differences were confirmed by comparative immunoprecipitation analysis of enriched membrane preparations with anti-beta-dystroglycan polyclonal antibody. These experiments revealed clear complex or non-complex formation between members of the dystrophin system, depending on the muscle type analyzed. Differences in the potential function of these various dystrophin complexes in fast or slow muscle fibers are discussed in relation to previous data obtained in corresponding mammalian tissues. (J Histochem Cytochem 49:857-865, 2001) PMID- 11410611 TI - Expression of cyclo-oxygenase-2 in naturally occurring squamous cell carcinomas in dogs. AB - Squamous cell carcinoma is one of the most common cancers in humans and is also a frequently diagnosed neoplasm in dogs. Induction of cyclo-oxygenase-2 (COX-2), a key rate-limiting enzyme in prostaglandin biosynthesis, has been implicated in the oncogenesis of various cancers in humans, including squamous cell carcinomas. However, expression of COX-2 has not been reported in spontaneous squamous cell carcinomas of non-human species. Canine squamous cell carcinomas share several similarities with the human disease. Therefore, the objective of this study was to determine whether COX isoenzymes were expressed in naturally occurring cases of squamous cell carcinomas in dogs. Canine normal skin (n=4) and squamous cell carcinomas (n=40) were studied by immunohistochemistry and immunoblotting analysis using polyclonal antibodies selective for COX-1 or COX-2. COX-2 was strongly expressed by neoplastic keratinocytes in all cases of squamous cell carcinomas, whereas no COX-2 was detected in normal skin and in the non neoplastic skin and oral mucosa included in the tumor tissue samples (p<0.01). Immunoblotting analysis confirmed the restricted expression of COX-2 (72,000- 74,000 molecular weight doublet) in squamous cell carcinomas only. In contrast, faint COX-1 staining was found in normal skin and in squamous cell carcinomas. This study demonstrates for the first time that COX-2 is induced in canine squamous cell carcinomas, and provides a new model to investigate the role and regulation of COX-2 gene expression in naturally occurring squamous cell carcinomas. (J Histochem Cytochem 49:867-875, 2001) PMID- 11410612 TI - Distribution of biglycan and decorin in collateral and cruciate ligaments and menisci of the rabbit knee joint. AB - The small leucine-rich proteoglycans (PGs) biglycan and decorin, and their mRNAs, have been localized during neonatal development and aging (3 weeks to 2 years) of collateral and cruciate ligaments and of menisci of the rabbit knee joint. In the collateral ligaments, biglycan and decorin are found between the bundles of collagen fibers at all ages. In cruciate ligaments the PGs are primarily around the cells. In neonatal ligaments all the cells express the mRNAs for biglycan and decorin, but in the collateral ligaments the number expressing the mRNAs is reduced at 8 months. In 3--week menisci the PGs are uniformly distributed in the matrix, but by 8 months biglycan is present primarily in the central fibrocartilaginous regions, whereas decorin is found peripherally. In neonates, all the cells express the mRNAs but the number is reduced in 8-month menisci. The results illustrate the precise localizations of biglycan and decorin in healthy rabbit ligaments and menisci which, after injury, must be reproduced in the repair tissue for normal strength to be regained. (J Histochem Cytochem 49:877 885, 2001) PMID- 11410613 TI - Expression and neural control of myogenic regulatory factor genes during regeneration of mouse soleus. AB - Given the importance of the myogenic regulatory factors (MRFs) for myoblast differentiation during development, the aims of this work were to clarify the spatial and temporal expression pattern of the four MRF mRNAs during soleus regeneration in mouse after cardiotoxin injury, using in situ hybridization, and to investigate the influence of innervation on the expression of each MRF during a complete degeneration/regeneration process. For this, we performed cardiotoxin injury-induced regeneration experiments on denervated soleus muscle. Myf-5, MyoD, and MRF4 mRNAs were detected in satellite cell-derived myoblasts in the first stages of muscle regeneration analyzed (2--3 days P-I). The Myf-5 transcript level dramatically decreased in young multinucleated myotubes, whereas MyoD and MRF4 transcripts were expressed persistently throughout the regeneration process. Myogenin mRNA was transiently expressed in forming myotubes. These results are discussed with regard to the potential relationships between MyoD and MRF4 in the satellite cell differentiation pathway. Muscle denervation precociously (at 8 days P-I) upregulated both the Myf-5 and the MRF4 mRNA levels, whereas the increase of both MyoD and myogenin mRNA levels was observed later, in the late stages of regeneration (30 days P-I). This significant accumulation of each differentially upregulated MRF during soleus regeneration after denervation suggests that each myogenic factor might have a distinct role in the regulatory control of muscle gene expression. This role is discussed in relation to the expression of the nerve-regulated genes, such as the nAChR subunit gene family. (J Histochem Cytochem 49:887-899, 2001) PMID- 11410614 TI - Microwave strategy for improving the simultaneous detection of estrogen receptor and galanin receptor mRNA in the rat hypothalamus. AB - In a attempt to improve the sensitivity of the simultaneous use of immunohistochemistry (IHC) with estrogen receptor (ER) and in situ hybridization (ISH) with a neuropeptide receptor, we first applied an existing microwave (MW) irradiation protocol for immunohistochemical detection of the estrogen receptor in frozen brain sections. Regions of interest were the preoptic area and the arcuate nucleus of the hypothalamus. ER signal was effective only after MW heating of sections in the two regions. Control sections without pretreatment exhibited no staining for ER. Second, the MW protocol was applied in a novel procedure that consists of evaluation of the expression of the galanin receptor mRNA with a radioactive riboprobe after MW pretreatment. The galanin receptor mRNA signal intensity obtained after heating was quantitatively at least as good or significantly increased according to the region, with no discernible loss of tissue morphology. Finally, we describe a novel application of MW pretreatment on the same frozen section processed with ER antibody and a radioactive galanin receptor riboprobe. The stainings for estrogen and galanin receptors were intense in many cells of the preoptic area, with very low background. These results show that both IHC and ISH can be significantly improved by subjecting frozen sections to MW heating before the double labeling. This approach may provide a potential method to answer the important question of whether or not estrogen has a direct action on the expression of a peptide receptor. (J Histochem Cytochem 49:901-910, 2001) PMID- 11410615 TI - Lysis of prostate carcinoma cells by trifunctional bispecific antibodies (alpha EpCAM x alpha CD3). AB - Bispecific monoclonal antibodies (bsAbs) are a promising immunotherapeutic option for treatment of cancer, especially in situations of minimal residual disease. The combination of an anti-CD3 and anti-tumor-associated antigen antibody redirects cytotoxic T-lymphocytes towards malignant cells. Using a trifunctional bispecific antibody against EpCAM x CD3, that additionally activates Fc gamma R(+) accessory cells via its Fc region, we investigated the interaction between three EpCAM(+) prostate carcinoma cell lines and peripheral blood mononuclear cells (PBMCs) of healthy donors and patients with prostate carcinoma (PC). Visualization was performed by double immunocytochemical methods and computerized sequential video microscopy. Tumor cells and PBMCs supplemented with alpha EpCAM x alpha CD3 in 16-well chamber slides resulted in lysis of tumor cells within 1- 3 days without any differences between patient and healthy donor PBMCs. The characteristic necrotic way of tumor cell killing (rounding, swelling, disrupting) could be observed in computerized sequences of video frames. Simultaneously, we could not reveal any form of apoptotic signal using three different apoptotic markers (TUNEL, M30 cyto death, anti-active caspase 3). Within the first 48 hr we observed typical PBMC cluster formation with increasing cell proliferation. PBMCs surrounding the tumor cells were not dominated by CD4(+), CD8(+), or CD14(+) cells. Lymphocytes with pore-forming perforin proteins concentrated towards the tumor target cells. Our combination of double immunocytochemical and computerized video microscopic techniques may serve as an important improvement of validity of cell-cell interaction experiments using in vitro models. (J Histochem Cytochem 49:911-917, 2001) PMID- 11410616 TI - Cytokeratin 20 immunoreactivity in renal oncocytomas. AB - Cytokeratins (CKs) are a group of 20 antigenically distinct intermediate filaments, generally confined to epithelia and their neoplasms. Immunostaining for CKs, in particular coordinate staining for CK7 and CK20, has become a useful tool in diagnostic pathology. Although studies defining CK distribution in neoplasms identify 0--7.7% of renal cell carcinomas (RCCs) positive for CK20, none has described the incidence of CK20 immunopositivity in renal oncocytomas (ROs). Distinction between RCC and RO may be difficult but this distinction is clinically significant, prompting us to establish the incidence of CK20 positivity in RO. We selected fifteen surgical cases of RO from our archives and studied their immunoreactivity for CKs including CK7 and CK20; 12/15 (80%) were positive for CK20, with variation in the number of cells staining. There was also variation in the distribution of CKs within the cells, including diffuse cytoplasmic, perinuclear, and a punctate or dot-like pattern. Such punctate staining corresponds to cytoplasmic balls of intermediate filaments and has been described with CAM 5.2 in RO and CK20 in Merkel cell carcinomas. Our findings suggest that CK20 immunohistochemistry is a useful tool for distinguishing RCCs from ROs. (J Histochem Cytochem 49:919-920, 2001) PMID- 11410617 TI - Differential staining of DNA strand breaks in dried comet assay slides. AB - The comet assay involves embedding cells in agarose on microscope slides. After lysis and electrophoresis, staining is usually performed with a fluorescent DNA binding dye and observation is carried out on fresh wet slides through an epifluorescence microscope. We present here a simple alternative for preservation of the agarose comet slides and a fluorescent staining that allows fine differential analysis of DNA strand breaks under confocal microscopy. Lymphocytes were processed according to previous published methods. Slides were quickly dehydrated in a hot oven at 50C for 20 min. Once the agarose layer was dried and reduced to a thin film, slides were treated with RNase. Image analysis showed higher tail length, total area, and tail moment. Using confocal microscopic optical sectioning, a thickness of approximately 180 microm for wet slides and 12 microm for dehydrated gels was calculated. Acridine orange, used for DNA differential staining, allowed quantitation of metachromasia and orthochromasia with confocal scanning microscopy. Differences between alkaline and neutral comet assay with AO were clear-cut and, in principle, a metachromatic index can be calculated. (J Histochem Cytochem 49:921-922, 2001) PMID- 11410618 TI - Protease XXIV increases detection of mucin gene expression during in situ hybridization in archival tissue. AB - Digoxigenin-labeled riboprobes of six groups of human mucins were evaluated for sensitivity in archival tissue, using protease XXIV or proteinase K during in situ hybridization. (J Histochem Cytochem 49:923-924, 2001) PMID- 11410619 TI - Monitoring signal transduction in cancer: from chips to fish. AB - The microarray format of RNA transcript analysis should provide new clues to carcinogenic processes. Because of the complex and heterogeneous nature of most tumor samples, histochemical techniques, particularly RNA fluorescent in situ hybridization (FISH), are required to test the predictions from microarray expression experiments. Here we describe our approach to verify new microarray data by examining RNA expression levels of five to seven different transcripts in a very few cells via FISH. (J Histochem Cytochem 49:925-926, 2001) PMID- 11410620 TI - Effect of the storage period of paraffin sections on the detection of mRNAs by in situ hybridization. AB - In this study we evaluated whether storing non-deparaffinized sections can affect the detection of specific mRNAs by radioactive in situ hybridization (ISH). Using a standard ISH protocol, we hybridized serial sections of paraffin blocks stored for different periods of time with (33)P-labeled riboprobes specific for rat Type III collagen and matrix metalloproteinase-2 (MMP-2). Signal intensities were evaluated using a phosphorimager and by blinded microscopic examination. For slides hybridized with the Type III collagen riboprobe, signal intensities measured with the phosphorimager or evaluated by microscopic examination were negatively correlated with the storage period of the sections. For slides hybridized with the MMP-2 riboprobe, differences in signal intensity could be detected, albeit inconsistently, with the phosphorimager, although microscopic examination consistently indicated stronger signals in freshly sectioned slides compared to slides stored for 2 weeks or more. We concluded that it was preferable to use recently prepared sections for trying to locate mRNAs in paraffin-embedded tissues by ISH. In addition, our results suggest that quantifying signal intensity using a phosphorimager is feasible for abundant mRNAs or when large differences in expression are anticipated.(J Histochem Cytochem 49:927-928, 2001) PMID- 11410621 TI - Comparison of apoptosis and terminal differentiation: the mammalian aging process. AB - Apoptosis is the ordered chain of events that lead to cell destruction. Terminal differentiation (denucleation) is the process in which cells lose their nuclei but remain functional. Our group examined cell death in three tissues using two different fixatives and a postfixation procedure, involving young (5 months) and old (2 years) guinea pigs. The data reveal that B-DNA and Z-DNA content decreases, whereas single-stranded (ss-) DNA increases, in older tissues undergoing apoptosis (skin and cornea) and terminal differentiation (ocular lens). We speculate that some of the factors that contribute to the aging process might also be responsible for the enhanced amount of damaged DNA in older tissues undergoing cell death. (J Histochem 49:929-930, 2001) PMID- 11410622 TI - Ion selectivities of the Ca(2+) sensors for exocytosis in rat phaeochromocytoma cells. AB - 1. The ion selectivities of the Ca(2+) sensors for the two components of exocytosis in rat phaeochromocytoma (PC12) cells were examined by measurement of membrane capacitance and amperometry. The cytosolic concentrations of metal ions were increased by photolysis of caged-Ca(2+) compounds and measured with low affinity indicators benzothiazole coumarin (BTC) or 5-nitrobenzothiazole coumarin (BTC-5N). 2. The Ca(2+)-induced increases in membrane capacitance comprised two phases with time constants of 30--100 ms and 5 s. Amperometric events reflecting the exocytosis of large dense-core vesicles occurred selectively in the slow phase, even with increases in the cytosolic Ca(2+) concentration of > 0.1 mM. 3. The slow component of exocytosis was activated by all metal ions investigated, including Cd(2+) (median effective concentration, 18 pM), Mn(2+) (500 nM), Co(2+) (900 nM), Ca(2+) (8 microM), Sr(2+) (180 microM), Ba(2+) (280 microM) and Mg(2+) (> 5 mM). In contrast, the fast component of exocytosis was activated by Cd(2+) (26 pM), Mn(2+) (620 nM), Ca(2+) (24 microM) and Sr(2+) (320 microM), but was only slightly increased by Ba(2+) (> 2 mM) and Co(2+) and not at all by Mg(2+). 4. The fast component, but not the slow component, was competitively blocked by Na(+) (median effective concentration, 44 mM) but not by Li(+), K(+) or Cs(+). Thus, the Ca(2+) sensor for the fast component of exocytosis is more selective than is that for the slow component; moreover, this selectivity appears to be based on ionic radius, with cations with radii of 0.84 to 1.13 A (1 A = 0.1 nm) being effective. 5. These data support a role for synaptotagmin--phospholipid as the Ca(2+) sensor for the exocytosis of large dense-core vesicles and they suggest that an additional Ca(2+)-sensing mechanism operates in the synchronous exocytosis of synaptic-like vesicles. PMID- 11410623 TI - Inhibition of creatine kinase reduces the rate of fatigue-induced decrease in tetanic [Ca(2+)](i) in mouse skeletal muscle. AB - 1. Ca(2+)-phosphate (P(i)) precipitation in the sarcoplasmic reticulum (SR) may cause reduced SR Ca(2+) release in skeletal muscle fatigue. To study this, we inhibited the creatine kinase (CK) reaction with 2,4-dinitro-1-fluorobenzene (DNFB). The hypothesis was that with inhibition of CK, phosphocreatine would not break down to creatine and P(i). Therefore P(i) transport into the SR would be limited and Ca(2+)-P(i) precipitation would not occur. 2. Intact single fibres from a mouse foot muscle were fatigued by repeated short tetani under control conditions or after exposure to DNFB (10 microM). The free myoplasmic concentrations of Ca(2+) ([Ca(2+)](i)) and Mg(2+) ([Mg(2+)](i)) were measured with indo-1 and mag-indo-1, respectively. Changes in [Mg(2+)](i) were assumed to reflect alterations in myoplasmic ATP concentration. 3. During the first 10 fatiguing tetani, tetanic [Ca(2+)](i) increased both in control and after DNFB exposure. Thereafter tetanic [Ca(2+)](i) fell and the rate of fall was about fourfold lower after DNFB exposure compared with control. 4. Under control conditions, there was a good relationship between declining tetanic [Ca(2+)](i) and increasing [Mg(2+)](i) during the final part of fatiguing stimulation. This correlation was lost after DNFB exposure. 5. In conclusion, the present data fit with a model where Ca(2+)-P(i) precipitation inhibits SR Ca(2+) release in fatigue produced by repeated short tetani. Furthermore, the results suggest that the rate of P(i) transport into the SR critically depends on the myoplasmic Mg(2+)/ATP concentration. PMID- 11410624 TI - Thiophosphorylation-induced Ca(2+) sensitization of guinea-pig ileum contractility is not mediated by Rho-associated kinase. AB - 1. Incubation of beta-escin-permeabilized guinea-pig longitudinal ileal smooth muscle with ATP gamma S under conditions that do not lead to thiophosphorylation of regulatory light chains of myosin (r-MLC) increased subsequent Ca(2+) sensitivity of force and r-MLC phosphorylation. In this study we tested whether this is due to activation of the Rho and/or Rho-associated kinase (ROK) as it is the case in agonist-induced Ca(2+) sensitization. 2. The increase in Ca(2+) sensitivity induced by pretreatment with ATP gamma S at pCa > 8 with the myosin light chain kinase (MLCK) inhibitor ML-9 in rigor solution was associated with (35)S incorporation into the regulatory subunit of myosin light chain phosphatase (MLCP), MYPT1, and several other high molecular mass proteins. No thiophosphorylation of r-MLC, MLCK, caldesmon, calponin and CPI-17 was detected. 3. While the relatively specific inhibitor of ROK, Y 27632, inhibited the carbachol-induced increase in Ca(2+) sensitivity with an IC(50) of 1.4 microM, the ATP gamma S-induced increase in Ca(2+) sensitivity and thiophosphorylation of MYPT1 was not inhibited. Inhibiton of Rho by exoenzyme C3 also had no effect. 4. Only staurosporine (2 microM), but not the PKC inhibitor peptide 19-31, nor genistein nor PD 98059, inhibited the ATP gamma S-induced Ca(2+) sensitization of force, r-MLC phosphorylation, and the (35)S incorporation into MYPT1. 5. The staurosporine-sensitive kinase(s) appeared to be tightly associated with the contractile apparatus because treatment of Triton-skinned preparations with ATP gamma S also induced a staurosporine-sensitive increase in Ca(2+) sensitivity of contraction. Since there was very little immunoreactivity with antibodies to p(21)-associated kinase (PAK) in Triton-skinned preparations, the staurosporine sensitive kinase most probably is not PAK. 6. GTP gamma S had an additive effect on ATP gamma S-induced sensitization at saturating concentrations of ATP gamma S. The additional effect of GTP gamma S was inhibited by Y 27632. 7. We conclude that treatment with ATP gamma S under ATP-free conditions, unmasks a staurosporine-sensitive kinase which induces a large increase in Ca(2+) sensitivity that is most likely to be due to thiophosphorylation of MYPT1. The kinase is distinct from ROK. The physiological significance of this kinase, which is tightly associated with the contractile apparatus, is not known at present. PMID- 11410625 TI - Glia-derived signals induce synapse formation in neurones of the rat central nervous system. AB - 1. To study the effects of glial cells on synapse formation, we established microcultures of purified rat retinal ganglion cells (RGCs) and monitored synapse (autapse) development in single neurones using electrophysiological recordings, FM1-43 labelling and immunocytochemistry. 2. Solitary neurones grew ramifying neurites, but formed only very few and inefficient excitatory autapses, when cultured for up to 2 weeks in defined medium and in the absence of glial cells. 3. Treatment of glia-free microcultures of RGCs with glia-conditioned medium (GCM) increased the number of autapses per neurone by up to 10-fold. This was indicated by a similar increase in the frequency of spontaneous events and the number of FM1-43-labelled functional release sites and of puncta, where pre- and postsynaptic markers colocalized. 4. In addition, GCM treatment enhanced the efficacy of presynaptic transmitter release as indicated by lower failure rates of stimulation-induced excitatory autaptic currents, a 200-fold increase in the frequency of asynchronous release and an accelerated stimulation-induced FM1-43 destaining. Furthermore, GCM induced an increase in the quantal size. 5. GCM affected autaptic activity not immediately, but with a delay of 24 h, and the effects on stimulation-induced autaptic currents occurred before changes in the frequency of spontaneous events indicating an early strengthening of existing autapses followed by a later increase in autapse number. 6. The observed effects were mediated by proteinase K-sensitive factors in GCM and occurred independently of electrical activity. 7. These results suggest that soluble glia-derived signals induce synapse formation and maturation in neurones of the central nervous system (CNS). PMID- 11410626 TI - Contribution of potential EF hand motifs to the calcium-dependent gating of a mouse brain large conductance, calcium-sensitive K(+) channel. AB - 1. The large conductance, calcium-sensitive K(+) channel (BK(Ca) channel) is a unique member of the K(+)-selective ion channel family in that activation is dependent upon both direct calcium binding and membrane depolarization. Calcium binding acts to dynamically shift voltage-dependent gating in a negative or left ward direction, thereby adjusting channel opening to changes in cellular membrane potential. 2. We hypothesized that the intrinsic calcium-binding site within the BK(Ca) channel alpha subunit may contain an EF hand motif, the most common, naturally occurring calcium binding structure. Following identification of six potential sites, we introduced a single amino acid substitution (D/E to N/Q or A) at the equivalent of the -z position of a bona fide EF hand that would be predicted to lower calcium binding affinity at each of the six sites. 3. Using macroscopic current recordings of wild-type and mutant BK(Ca) channels in excised inside-out membrane patches from HEK 293 cells, we observed that a single point mutation in the C-terminus (Site 6, FLD(923)QD to N), adjacent to the 'calcium bowl' described by Salkoff and colleagues, shifted calcium-sensitive gating right ward by 50--65 mV over the range of 2--12 microM free calcium, but had little effect on voltage-dependent gating in the absence of calcium. Combining this mutation at Site 6 with a similar mutation at Site 1 (PVD(81)EK to N) in the N terminus produced a greater shift (70--90 mV) in calcium-sensitive gating over the same range of calcium. We calculated that these combined mutations decreased the apparent calcium binding affinity approximately 11-fold (129.5 microM vs. 11.3 microm) compared to the wild-type channel. 4. We further observed that a bacterially expressed protein encompassing Site 6 of the BK(Ca) channel C terminus and bovine brain calmodulin were both able to directly bind (45)Ca(2+) following denaturation and polyacrylamide gel electrophoresis (e.g. SDS-PAGE). 5. Our results suggest that two regions within the mammalian BK(Ca) channel alpha subunit, with sequence similarities to an EF hand motif, functionally contribute to the calcium-sensitive gating of this channel. PMID- 11410627 TI - The consequences of disrupting cardiac inwardly rectifying K(+) current (I(K1)) as revealed by the targeted deletion of the murine Kir2.1 and Kir2.2 genes. AB - 1. Ventricular myocytes demonstrate a steeply inwardly rectifying K(+) current termed I(K1). We investigated the molecular basis for murine I(K1) by removing the genes encoding Kir2.1 and Kir2.2. The physiological consequences of the loss of these genes were studied in newborn animals because mice lacking Kir2.1 have a cleft palate and die shortly after birth. 2. Kir2.1 (-/-) ventricular myocytes lack detectable I(K1) in whole-cell recordings in 4 mM external K(+). In 60 mM external K(+) a small, slower, residual current is observed. Thus Kir2.1 is the major determinant of I(K1). Sustained outward K(+) currents and Ba(2+) currents through L- and T-type channels were not significantly altered by the mutation. A 50 % reduction in I(K1) was observed in Kir2.2 (-/-) mice, raising the possibility that Kir2.2 can also contribute to the native I(K1). 3. Kir2.1 (-/-) myocytes showed significantly broader action potentials and more frequent spontaneous action potentials than wild-type myocytes. 4. In electrocardiograms of Kir2.1 (-/-) neonates, neither ectopic beats nor re-entry arrhythmias were observed. Thus the increased automaticity and prolonged action potential of the mutant ventricular myocytes were not sufficiently severe to disrupt the sinus pacing of the heart. The Kir2.1 (-/-) mice, however, had consistently slower heart rates and this phenotype is likely to arise indirectly from the influence of Kir2.1 outside the heart. 5. Thus Kir2.1 is the major component of murine I(K1) and the Kir2.1 (-/-) mouse provides a model in which the functional consequences of removing I(K1) can be studied at both cellular and organismal levels. PMID- 11410628 TI - GABA-evoked chloride currents do not differ between dendrites and somata of rat neocortical neurons. AB - 1. We performed patch-clamp recordings on acutely isolated somata and dendritic segments of rat neocortical neurons, in order to compare the reversal potential (E(GABA)) and relative density of GABA(A) receptor-mediated Cl(-) currents in these two cellular compartments. 2. Currents were recorded with the Cl(-) impermeable pore former gramicidin (25--75 microg ml(-1)) in HCO(3)(-)-free bath solution. Voltage ramps (-110 to -30 mV) from a holding potential (V(h)) of -60 mV in the absence and presence of 2 microM GABA were used to construct instantaneous current-voltage relationships. Currents were abolished by co application of GABA with the GABA(A) receptor antagonist bicuculline (40 microM). 3. GABA conductance, normalized to membrane surface area, was not different in somata and dendrites. In addition, E(GABA) was not different in the two compartments. 4. Replacement of intracellular K(+) with Cs(+) resulted in a significantly more depolarized E(GABA) in both somata and dendrites. These results suggest that the resting intracellular Cl(-) concentration ([Cl(-)](i)) is similar in somata and dendrites and that an outward Cl(-) transporter system maintains low [Cl(-)](i). PMID- 11410629 TI - The effects of capsaicin and acidity on currents generated by noxious heat in cultured neonatal rat dorsal root ganglion neurones. AB - 1. The effects of capsaicin, acidic pH, ATP, kainate and GABA on currents generated by noxious heat were studied in cultured dorsal root ganglion (DRG) neurones (< 20 microm in diameter) isolated from neonatal rats. The patch clamp technique was used to record membrane currents or changes of membrane potential. 2. In agreement with previous results, inward membrane currents (I(heat)) induced by a 3 s ramp of increasing temperature from room temperature (approximately 23 degrees C) to over 42 degrees C varied greatly between cells (-100 pA to -2.4 nA at 48 degrees C) and had a temperature coefficient (Q(10)) > 10 over the range of 43-52 degrees C. 3. Capsaicin potentiated the heat-induced current even when capsaicin, at room temperature, had little or no effect on its own. In cells in which capsaicin induced no or very small membrane current at room temperature (< 50 pA), I(heat) exhibited detectable activation above 40 degrees C and increased 5.1 +/- 1.1 (n = 37) and 6.3 +/- 2.0 (n = 18) times at 0.3 and 1 microM capsaicin, respectively. 4. A rapid decrease in extracellular pH from 7.3 to 6.8, 6.3 or 6.1 produced an inward current which inactivated in ~5 s either completely (pH 6.8 or 6.3) or leaving a small current (approximately 50 pA) for more than 2 min (pH 6.1). After inactivation of the initial low pH-induced current, I(heat) at 48 degrees C increased 2.3 +/- 0.4 times at pH 6.8, 4.0 +/- 0.6 times at pH 6.3 and 4.8 +/- 0.8 times at pH 6.1 with a Q(10) > 10 (n = 16). 5. ATP (n = 22), kainate (n = 7) and GABA (n = 8) at 100 microM, produced an inactivating inward current in all heat-sensitive DRG neurones tested. During inactivation and in the presence of the drug, I(heat) was increased slightly with ATP and unaffected with kainate and GABA. These agents apparently do not directly affect the noxious heat receptor. 6. The results indicate a novel class of capsaicin-sensitive cells, in which capsaicin evokes no or very small inward current but nevertheless increases sensitivity to noxious heat. PMID- 11410630 TI - Free intracellular Mg(2+) concentration and inhibition of NMDA responses in cultured rat neurons. AB - 1. Intracellular Mg(2+) (Mg(2+)(i)) blocks single-channel currents and modulates the gating kinetics of NMDA receptors. However, previous data suggested that Mg(2+)(i) inhibits whole-cell current less effectively than predicted from excised-patch measurements. We examined the basis of this discrepancy by testing three hypothetical explanations. 2. To test the first hypothesis, that control of free Mg(2+)(i) concentration ([Mg(2+)](i)) during whole-cell recording was inadequate, we measured [Mg(2+)](i) using mag-indo-1 microfluorometry. The [Mg(2+)](i) measured in cultured neurons during whole-cell recording was similar to the pipette [Mg(2+)] measured in vitro, suggesting that [Mg(2+)](i) was adequately controlled. 3. To test the second hypothesis, that open-channel block by Mg(2+)(i) was modified by patch excision, we characterised the effects of Mg(2+)(i) using cell-attached recordings. We found the affinity and voltage dependence of open-channel block by Mg(2+)(i) similar in cell-attached and outside-out patches. Thus, the difference between Mg(2+)(i) inhibition of whole cell and of patch currents cannot be attributed to a difference in Mg(2+)(i) block of single-channel current. 4. The third hypothesis tested was that the effect of Mg(2+)(i) on channel gating was modified by patch excision. Results of cell-attached recording and modelling of whole-cell data suggest that the Mg(2+)(i)-induced stabilisation of the channel open state is four times weaker after patch excision than in intact cells. This differential effect of Mg(2+)(i) on channel gating explains why Mg(2+)(i) inhibits whole-cell NMDA responses less effectively than patch responses. PMID- 11410631 TI - NMDA receptor- and metabotropic glutamate receptor-dependent synaptic plasticity induced by high frequency stimulation in the rat dentate gyrus in vitro. AB - 1. The mechanisms of long-term potentiation (LTP) and long-term depression (LTD) induced by brief high frequency stimulation (HFS), paired with a particular pattern and amplitude of depolarisation has been investigated in the medial perforant pathway of the dentate gyrus of the 2- to 3-week-old rat hippocampus in vitro. 2. N-Methyl-D-aspartate (NMDA) receptor (NMDAR) activation was measured quantitatively during HFS-induced NMDAR-dependent LTP, LTD and at the LTD--LTP crossover point in order to test the hypothesis that the induction of the particular form of plasticity depends on the intensity of NMDAR activation. 3. The induction of LTD, the LTD--LTP crossover point and LTP was associated with an increasing NMDAR charge transfer. 4. In addition to the NMDAR-dependent LTD, a group I metabotropic glutamate receptor (mGluR)-dependent LTD could be induced by high intensity HFS paired with depolarisation under conditions of NMDAR inhibition. 5. The induction of mGluR-dependent LTD requires membrane depolarisation, Ca(2+) influx via L-type Ca(2+) channels and a rise in intracellular Ca(2+). 6. Quantal analysis involving minimal stimulation demonstrated that the mGluR-dependent LTD induction was associated with a decrease in potency and an increase in failure rate. PMID- 11410632 TI - Photolysis-induced suppression of inhibition in rat hippocampal CA1 pyramidal neurons. AB - 1. Whole cell patch clamp recording, Ca(2+) measurement with ratiometric fluorescent dyes and photolysis of caged Ca(2+) were combined to investigate the depolarization- and photolysis-induced suppression of inhibition (DSI and PSI) in rat hippocampal CA1 pyramidal cells. 2. A 5-s depolarization from -70 mV to 0 mV or a 6-s photolysis of nitrophenyl-EGTA (NPE) in cell bodies could each depress the frequency of spontaneous inhibitory postsynaptic currents (IPSCs) and the amplitude of evoked IPSCs while elevating intracellular Ca(2+) concentration ([Ca(2+)](i)). 3. Within a cell the elevation of [Ca(2+)](i) induced by depolarization was inversely related to that induced by photolysis, suggesting that higher [NPE] is more effective in releasing caged Ca(2+) but also increases buffer capacity to reduce [Ca(2+)](i) rises caused by Ca(2+) influx through voltage-dependent Ca(2+) channels. 4. Both DSI and PSI were linearly related to [Ca(2+)](i), with a 50 % reduction in transmission occurring at about 3.6--3.9 microM. 5. [Ca(2+)](i) recovered more quickly than DSI, indicating that the duration of DSI is not set simply by the duration of [Ca(2+)](i) elevation, but rather entails other rate-limiting processes. 6. We conclude that DSI is activated by micromolar [Ca(2+)](i) acting far from sites of Ca(2+) entry through channels in the plasma membrane. PMID- 11410633 TI - The conductance underlying the parallel fibre slow EPSP in rat cerebellar Purkinje neurones studied with photolytic release of L-glutamate. AB - 1. Tetanic stimulation of parallel fibres (PFs) produces a slow EPSP (sEPSP) or slow EPSC (sEPSC) in Purkinje neurones (PNs), mediated by type 1 metabotropic glutamate receptors (mGluR1). The conductance change underlying the sEPSP was investigated with rapid photolytic release of L-glutamate from nitroindolinyl (NI)-caged glutamate with ionotropic glutamate receptors blocked, and showed a slow mGluR1-activated cation channel. 2. In cerebellar slices rapid photolytic release (t (1/2) < 0.7 ms) of 7--70 microM L-glutamate on PNs voltage clamped at 65 mV activated first a transient inward current, peaking in 8 ms, followed by a slow inward current with time course similar to the PF sEPSP, peaking at -1 nA in 700 ms. 3. The initial current was inhibited by 300 microM threo-hydroxyaspartate (THA) and did not reverse as the potential was made positive up to +50 mV, suggesting activation of electrogenic glutamate uptake. 4. The slow current was inhibited reversibly by 1 mM (R,S)-MCPG or the non-competitive mGluR1 antagonist CPCCOEt (20 microM), indicating activation of metabotropic type 1 glutamate receptors. The mGluR current was associated with increases of input conductance and membrane current noise, and reversed close to 0 mV, indicating activation of channels permeant to Na(+) and K(+). 5. The sEPSC was not blocked by Cd(2+), Co(2+), Mg(2+) or Gd(3+) ions, by the inhibitor of hyperpolarisation-activated current (I(H)) ZD7288, or by the purinoceptor inhibitor PPADS. Activation was not affected by inhibitors of phospholipase C (PLC) or protein kinase C (PKC), nor mimicked by photorelease of InsP(3) or Ca(2+). The results show that mGluR1 in PNs produces a slow activation of cation-permeable ion channels which is not mediated by PLC activation, Ca(2+) release from stores, or via the activation of PKC. PMID- 11410634 TI - Coupling and uncoupling of activity-dependent increases of neuronal activity and blood flow in rat somatosensory cortex. AB - 1. Electrical stimulation of the infraorbital nerve was used to examine the coupling between neuronal activity and cerebral blood flow (CBF) in rat somatosensory cortex by laser Doppler flowmetry and extracellular recordings of field potentials. 2. The relationship between field potential (FP) and CBF amplitudes was examined as a function of the stimulus intensity (0--2.0 mA) at fixed frequency (3 Hz). FP amplitudes up to 2.0-2.5 mV were unaccompanied by increases of CBF. Above this threshold, CBF and FP amplitudes increased proportionally. 3. At fixed stimulus intensity of 1.5 mA, CBF increases were highly correlated to FP amplitudes at low frequencies of stimulation (< 2 Hz), but uncoupling was observed at stimulation frequencies of 2--5 Hz. The evoked responses were independent of stimulus duration (8--32 s). 4. The first rise in CBF occurred within the first 0.2 s after onset of stimulation in the upper 0- 250 microm of the cortex. Latencies were longer (1.0--1.2 s) in lower cortical layers in which CBF and FP amplitudes were larger. 5. Local AMPA receptor blockade attenuated CBF and FP amplitudes proportionally. 6. This study showed that activity-dependent increases in neuronal activity and CBF were linearly coupled under defined conditions, but neuronal activity was well developed before CBF started to increase. Consequently, the absence of a rise in CBF does not exclude the presence of significant neuronal activity. The CBF increase in upper cortical layers preceded the rise in lower layers suggesting that vessels close to or at the brain surface are the first to react to neuronal activity. The activity-dependent rise in CBF was explained by postsynaptic activity in glutamatergic neurons. PMID- 11410635 TI - Simultaneous intracellular recordings from longitudinal and circular muscle during the peristaltic reflex in guinea-pig distal colon. AB - 1. Simultaneous intracellular recordings were made from longitudinal muscle (LM) and circular muscle (CM) cells of guinea-pig distal colon during the peristaltic reflex. 2. Spontaneous rhythmical depolarizations with superimposed action potentials (mean amplitude: 19 +/- 2 mV) were regularly recorded from the LM (mean interval: 7 +/- 1 s). In contrast, in the CM layer, spontaneous action potentials occurred with an irregular frequency. Although spontaneous action potentials in LM were rarely correlated in time with those in CM, spontaneous inhibitory junction potentials (sIJPs) were found to occur synchronously in both muscles (5 out of 27 animals; 19 %). 3. Graded inflation of an intra-luminal balloon or mucosal stimulation oral to the recording electrodes elicited gradeable compound IJPs synchronously in both LM (mean amplitude: 6 +/- 1 mV) and CM (mean amplitude: 9 +/- 1 mV) (descending inhibitory reflex). Evoked IJPs were often followed by action potentials in both muscle layers. 4. Mucosal stimuli applied anal to the recording electrodes elicited compound excitatory junction potentials (EJPs) synchronously in both muscles layers that were often associated with the generation of action potentials. In the LM, evoked EJP amplitudes ranged from 3 mV (subthreshold) to 31 mV (including the action potential) and in the CM from 4 mV (subthreshold) to 44 mV (including the action potential). 5. Apamin (500 nM) reduced the evoked IJP in the CM by 55 % (from 11 +/- 2 to 5 +/- 1 mV), but caused no significant reduction in the LM layer (from 8 +/- 1 to 6 +/- 1 mV). Apamin-resistant IJPs in both muscle layers were likely to be due to nitric oxide, since they were abolished by L-NA (100 microM). 6. Atropine (1 microM) abolished the ascending excitatory reflex in both muscles. 7. Injection of neurobiotin into the LM and CM confirmed that simultaneous intracellular recordings were made from different muscle layers. 8. In conclusion, during the peristaltic reflex, the LM and CM layers receive synchronous inhibitory neuromuscular inputs during descending inhibition and synchronous excitatory neuromuscular inputs during ascending excitation. No evidence was found to support reciprocal innervation. PMID- 11410636 TI - Gastric distension-induced pyloric relaxation: central nervous system regulation and effects of acute hyperglycaemia in the rat. AB - 1. The pylorus plays an important role in the regulation of gastric emptying. In addition to the autonomic neuropathy associated with long-standing diabetes, acute hyperglycaemia per se has effects on gastric emptying. In this study, the role of the central nervous system in modulating the effects of hyperglycaemia on gastric distension-induced pyloric relaxation was investigated. 2. Gastric distension-induced pyloric relaxation was significantly reduced by subdiaphragmatic vagotomy, hexamethonium (20 mg kg(-1)) and N (G)-nitro-L arginine methyl ester (L-NAME; 10 mg kg(-1)), a nitric oxide synthase (NOS) biosynthesis inhibitor, in anaesthetized rats. In contrast, neither splanchnectomy nor guanethidine (5 mg kg(-1)) had an effect. 3. An intravenous (I.V.) infusion of D-glucose (20 %) for 30 min, which increased blood glucose concentrations from 5.4 to 12.8 mM, significantly inhibited gastric distension induced pyloric relaxation. 4. An intracerebroventricular (I.C.V.) injection of D glucose (3 micromol) also significantly inhibited gastric distension-induced pyloric relaxation without affecting peripheral blood glucose concentrations. 5. I.V. infusion of D-glucose significantly elevated hypothalamic neuropeptide Y (NPY) concentrations. 6. Intracerebroventricular (I.C.V.) administration of NPY (0.03--3 nmol) and a Y1 receptor agonist, [leu(31), pro(34)] NPY (0.03--3 nmol), significantly inhibited gastric distension-induced pyloric relaxation in a dose dependent manner. 7. I.C.V. administration of a Y1 receptor antagonist, BIBP 3226 (30 nmol), and of a NPY antibody (titre 1:24 000, 3 microl) abolished the inhibitory effects of hyperglycaemia on gastric distension-induced pyloric relaxation. 8. Taken together, these findings suggest that gastric distension induced pyloric relaxation is mediated via a vago-vagal reflex and NO release. Acute hyperglycaemia stimulates hypothalamic NPY release, which, acting through the Y1 receptor, inhibits gastric distension-induced pyloric relaxation in rats exposed to acute elevations in blood glucose concentrations. PMID- 11410637 TI - Abnormal aortic fatty acid composition and small artery function in offspring of rats fed a high fat diet in pregnancy. AB - 1. Disturbances of the in utero environment are associated with an increased risk of cardiovascular disease in adulthood. In this study we have determined whether abnormal vascular function in the adult offspring of rats fed a high saturated fat diet in pregnancy is associated with altered plasma lipids or vascular fatty acid content. 2. Female Sprague-Dawley rats were fed a breeding diet (4 % fat) or a diet high in saturated fat (20 % fat) for 10 days prior to and throughout pregnancy, and during weaning. Female offspring were then fed a maintenance diet (3 % fat) until 160 days of age. 3. Endothelium-dependent relaxation induced by acetylcholine was blunted in isolated branches of the femoral artery from 160-day old female offspring of dams fed the saturated fat diet when compared with female offspring of dams fed the breeding diet. These offspring exhibited elevated plasma triglyceride and reduced plasma high density lipoprotein cholesterol concentrations. 4. The fatty acid composition of the aortas was abnormal, with a marked reduction in the content of arachidonic and docosahexaenoic acids. 5. This study demonstrates that a high fat diet in pregnant rats produces abnormal vascular function, plasma lipid disturbances and altered vascular fatty acid content in their female offspring during adulthood. PMID- 11410638 TI - Identification of higher brain centres that may encode the cardiorespiratory response to exercise in humans. AB - 1. Positron emission tomography (PET) was used to identify the neuroanatomical correlates underlying 'central command' during imagination of exercise under hypnosis, in order to uncouple central command from peripheral feedback. 2. Three cognitive conditions were used: condition I, imagination of freewheeling downhill on a bicycle (no change in heart rate, HR, or ventilation, V(I)): condition II, imagination of exercise, cycling uphill (increased HR by 12 % and V(I) by 30 % of the actual exercise response): condition III, volitionally driven hyperventilation to match that achieved in condition II (no change in HR). 3. Subtraction methodology created contrast A (II minus I) highlighting cerebral areas involved in the imagination of exercise and contrast B (III minus I) highlighting areas activated in the direct volitional control of breathing (n = 4 for both; 8 scans per subject). End-tidal P(CO(2)) (P(ET,CO(2))) was held constant throughout PET scanning. 4. In contrast A, significant activations were seen in the right dorso-lateral prefrontal cortex, supplementary motor areas (SMA), the right premotor area (PMA), superolateral sensorimotor areas, thalamus, and bilaterally in the cerebellum. In contrast B, significant activations were present in the SMA and in lateral sensorimotor cortical areas. The SMA/PMA, dorso lateral prefrontal cortex and the cerebellum are concerned with volitional/motor control, including that of the respiratory muscles. 5. The neuroanatomical areas activated suggest that a significant component of the respiratory response to 'exercise', in the absence of both movement feedback and an increase in CO(2) production, can be generated by what appears to be a behavioural response. PMID- 11410639 TI - Modulation of the Ca(2+)-induced Ca(2+) release cascade by beta-adrenergic stimulation in rat ventricular myocytes. AB - 1. To define the sub-cellular mechanisms of modulation of cardiac excitation contraction (E-C) coupling by the beta-adrenergic pathway, we carried out confocal Ca(2+) imaging in conjunction with recordings of inward Ca(2+) current in fluo-3-loaded patch-clamped rat ventricular myocytes. 2. Isoproterenol (isoprenaline; ISO) increased the amplitude of the inward Ca(2+) current and the globally measured intracellular Ca(2+) transients. The gain of calcium-induced calcium release (CICR) was increased at all membrane potentials but especially at positive membrane potentials (> +30 mV). ISO dramatically broadened the bell shaped voltage dependence of intracellular Ca(2+) transients by shifting the descending portion of the curve to very high membrane potentials. 3. The number of local release events (solitary sparks and conglomerates of overlapping sparks) induced by depolarizing steps to +30 mV was increased significantly by ISO. This potentiation of events was due to increased trigger calcium current (I(Ca)) as well the enhanced ability of I(Ca) to induce release. The amplitude and duration of solitary sparks were increased in the presence of ISO. In addition, ISO dramatically increased the proportion and the size ('mass') of clustered events. 4. Exclusion of Na(+) from the intra- and extracellular solutions prevented ISO from enhancing the ability of I(Ca) to trigger sparks. 5. We conclude that beta adrenergic stimulation enhances the gain of the CICR cascade by increasing the fidelity of dihydropyridine receptor (DHPR)--ryanodine receptor (RyR) coupling and by promoting cross-activation of RyRs in neighbouring release sites. Reverse Na(+)--Ca(2+) exchange (NCX) appears to play a role in the beta-adrenergic enhancement of CICR by effectively contributing to the Ca(2+) trigger. PMID- 11410640 TI - Exercise increases blood flow to locomotor, vestibular, cardiorespiratory and visual regions of the brain in miniature swine. AB - 1. The purpose of these experiments was to use radiolabelled microspheres to measure blood flow distribution within the brain, and in particular to areas associated with motor function, maintenance of equilibrium, cardiorespiratory control, vision, hearing and smell, at rest and during exercise in miniature swine. Exercise consisted of steady-state treadmill running at intensities eliciting 70 and 100 % maximal oxygen consumption (V(O(2),max)). 2. Mean arterial pressure was elevated by 17 and 26 % above that at rest during exercise at 70 and 100 % V(O(2),max), respectively. 3. Mean brain blood flow increased 24 and 25 % at 70 and 100 % V(O(2),max), respectively. Blood flow was not locally elevated to cortical regions associated with motor and somatosensory functions during exercise, but was increased to several subcortical areas that are involved in the control of locomotion. 4. Exercise elevated perfusion and diminished vascular resistance in several regions of the brain related to the maintenance of equilibrium (vestibular nuclear area, cerebellar ventral vermis and floccular lobe), cardiorespiratory control (medulla and pons), and vision (dorsal occipital cortex, superior colliculi and lateral geniculate body). Conversely, blood flow to regions related to hearing (cochlear nuclei, inferior colliculi and temporal cortex) and smell (olfactory bulbs and rhinencephalon) were unaltered by exercise and associated with increases in vascular resistance. 5. The data indicate that blood flow increases as a function of exercise intensity to several areas of the brain associated with integrating sensory input and motor output (anterior and dorsal cerebellar vermis) and the maintenance of equilibrium (vestibular nuclei). Additionally, there was an intensity-dependent decrease of vascular resistance in the dorsal cerebellar vermis. PMID- 11410641 TI - Effects of partial neuromuscular blockade on carotid baroreflex function during exercise in humans. AB - 1. This investigation was designed to determine the contribution of central command to the resetting of the carotid baroreflex during static and dynamic exercise in humans. 2. Thirteen subjects performed 3.5 min of static one-legged exercise (20 % maximal voluntary contraction) and 7 min dynamic cycling (20 % maximal oxygen uptake) under two conditions: control (no intervention) and with partial neuromuscular blockade (to increase central command influence) using Norcuron (curare). Carotid baroreflex function was determined at rest and during steady-state exercise using a rapid neck pressure/neck suction technique. Whole body Norcuron was repeatedly administered to effectively reduce hand-grip strength by approximately 50 % of control. 3. Partial neuromuscular blockade increased heart rate, mean arterial pressure, perceived exertion, lactate concentration and plasma noradrenaline concentration during both static and dynamic exercise when compared to control (P < 0.05). No effect was seen at rest. Carotid baroreflex resetting was augmented from control static and dynamic exercise by partial neuromuscular blockade without alterations in gain (P < 0.05). In addition, the operating point of the reflex was relocated away from the centring point (i.e. closer to threshold) during exercise by partial neuromuscular blockade (P < 0.05). 4. These findings suggest that central command actively resets the carotid baroreflex during dynamic and static exercise. PMID- 11410642 TI - Effects of exercise pressor reflex activation on carotid baroreflex function during exercise in humans. AB - 1. This investigation was designed to determine the contribution of the exercise pressor reflex to the resetting of the carotid baroreflex during exercise. 2. Ten subjects performed 3.5 min of static one-legged exercise (20 % maximal voluntary contraction) and 7 min dynamic cycling (20 % maximal oxygen uptake) under two conditions: control (no intervention) and with the application of medical anti shock (MAS) trousers inflated to 100 mmHg (to activate the exercise pressor reflex). Carotid baroreflex function was determined at rest and during exercise using a rapid neck pressure/neck suction technique. 3. During exercise, the application of MAS trousers (MAS condition) increased mean arterial pressure (MAP), plasma noradrenaline concentration (dynamic exercise only) and perceived exertion (dynamic exercise only) when compared to control (P < 0.05). No effect of the MAS condition was evident at rest. The MAS condition had no effect on heart rate (HR), plasma lactate and adrenaline concentrations or oxygen uptake at rest and during exercise. The carotid baroreflex stimulus-response curve was reset upward on the response arm and rightward to a higher operating pressure by control exercise without alterations in gain. Activation of the exercise pressor reflex by MAS trousers further reset carotid baroreflex control of MAP, as indicated by the upward and rightward relocation of the curve. However, carotid baroreflex control of HR was only shifted rightward to higher operating pressures by MAS trousers. The sensitivity of the carotid baroreflex was unaltered by exercise pressor reflex activation. 4. These findings suggest that during dynamic and static exercise the exercise pressor reflex is capable of actively resetting carotid baroreflex control of mean arterial pressure; however, it would appear only to modulate carotid baroreflex control of heart rate. PMID- 11410643 TI - Myoadenylate deaminase deficiency does not affect muscle anaplerosis during exhaustive exercise in humans. AB - 1. Myoadenylate deaminase (AMPD) deficiency is present in 1--2 % of the population. In theory, this deficiency may alter exercise energy metabolism by impairing the purine nucleotide cycle (PNC) and reducing tricarboxylic acid (TCA) cycle anaplerosis. The role of the PNC in TCA cycle anaplerosis is still a debated issue in physiology. Using patients with the AMPD1 mutation will allow a human 'knockout' approach to answering this question. 2. Muscle AMPD activity and genotype (whole blood AMPD1 analysis) was used to classify participants into three groups: n = 3 with absence of AMPD activity and -/- AMPD1 genotype (homozygous); n = 4 with less than 50 % normal AMPD activity and +/- genotype (heterozygous) and n = 12 with normal AMPD activity and +/+ genotype (control). Biopsies were taken from the vastus lateralis muscle before and after incremental cycle ergometry exercise to exhaustion. The muscle biopsies were analysed for AMPD activity, purine nucleotides/nucleosides and bases, creatine, phosphocreatine, amino acids, and the TCA cycle intermediates malate, citrate and fumarate. 3. Time to exhaustion on the cycle ergometer was not different between groups. Muscle adenosine monophosphate increased significantly with exercise for homozygous subjects as compared with the other groups (P < 0.05). Inosine monophosphate increased significantly after exercise for control (P < 0.05) but not for the homozygous subjects. There were no other between-group differences for any other measured variables. 4. In summary, complete and partial muscle AMPD deficiency did not affect TCA cycle anaplerosis, phosphocreatine hydrolysis, energy charge or exercise performance. PMID- 11410644 TI - Influence of exercise intensity on the on- and off-transient kinetics of pulmonary oxygen uptake in humans. AB - 1. The maximal oxygen uptake (V(O(2),peak)) during dynamic muscular exercise is commonly taken as a crucial determinant of the ability to sustain high-intensity exercise. Considerably less attention, however, has been given to the rate at which V(O(2)) increases to attain this maximum (or to its submaximal requirement), and even less to the kinetic features of the response following exercise. 2. Six, healthy, male volunteers (aged 22 to 58 years), each performed 13 exercise tests: initial ramp-incremental cycle ergometry to the limit of tolerance and subsequently, on different days, three bouts of square-wave exercise each at moderate, heavy, very heavy and severe intensities. Pulmonary gas exchange variables were determined breath by breath throughout exercise and recovery from the continuous monitoring of respired volumes (turbine) and gas concentrations (mass spectrometer). 3. For moderate exercise, the V(O(2)) kinetics were well described by a simple mono-exponential function, following a short cardiodynamic phase, with the on- and off-transients having similar time constants (tau(1)); i.e. tau(1,on) averaged 33 +/- 16 s (+/- S.D.) and tau(1,off) 29 +/- 6 s. 4. The on-transient V(O(2)) kinetics were more complex for heavy exercise. The inclusion of a second slow and delayed exponential component provided an adequate description of the response; i.e. tau(1,on) = 32 +/- 17 s and tau(2,on) = 170 +/- 49 s. The off-transient V(O(2)) kinetics, however, remained mono-exponential (tau(1,off) = 42 +/- 11 s). 5. For very heavy exercise, the on-transient V(O(2)) kinetics were also well described by a double exponential function (tau(1,on) = 34 +/- 11 s and tau(2,on) = 163 +/- 46 s). However, a double exponential, with no delay, was required to characterise the off-transient kinetics (i.e. tau(1,off) = 33 +/- 5 s and tau(2,off) = 460 +/- 123 s). 6. At the highest intensity (severe), the on-transient V(O(2)) kinetics reverted to a mono-exponential profile (tau(1,on) = 34 +/- 7 s), while the off transient kinetics retained a two-component form (tau(1,off) = 35 +/- 11 s and tau(2,off) = 539 +/- 379 s). 7. We therefore conclude that the kinetics of V(O(2)) during dynamic muscular exercise are strikingly influenced by the exercise intensity, both with respect to model order and to dynamic asymmetries between the on- and off-transient responses. PMID- 11410645 TI - Corticospinal excitation of presumed cervical propriospinal neurones and its reversal to inhibition in humans. AB - 1. This study addresses whether in human subjects indirect corticospinal excitation of upper limb motoneurones (MNs) relayed through presumed cervical propriospinal neurones (PNs) is paralleled by corticospinal activation of inhibitory projections to these premotoneurones. 2. The responses to transcranial magnetic stimulation (TMS), whether assessed as the compound motor-evoked potential (MEP) or the peak of corticospinal excitation elicited in the post stimulus time histograms (PSTHs) of single motor units, were conditioned by weak volleys to musculo-cutaneous, ulnar and superficial radial nerves. 3. Afferent volleys, which hardly modified the H reflex, significantly facilitated the corticospinal response produced by weak TMS. In PSTHs, the central delay of the peripheral facilitation of the peak of corticospinal excitation in MNs located at either end of the cervical enlargement was longer the more caudal the MN pool, suggesting an interaction in premotoneurones located rostral to the tested MNs. 4. Small increases in the strength of TMS (approximately 2--5 % of the maximal stimulator output) caused the facilitation to disappear and then to be reversed to inhibition. The facilitatory and inhibitory effects had the same latencies and spared the initial 0.5--1 ms of the corticospinal excitatory response. Both effects were more readily demonstrable when there was a co-contraction of the target muscle and the muscle innervated by nerve used for the conditioning stimulus. 5. The above features suggest that the inhibition resulted from disfacilitation due to suppression of corticospinal excitation passing through the presumed premotoneuronal relay. The reversal of the facilitation to inhibition by stronger corticospinal volleys is consistent with a well-developed system of 'feedback inhibitory interneurones' activated by corticospinal and afferent inputs inhibiting the presumed propriospinal excitatory premotoneurones. 6. It is argued that these findings might explain why simply stimulating the pyramidal tract or the motor cortex would fail to demonstrate this indirect corticospinal projection in the macaque monkey and in humans. PMID- 11410649 TI - Survey and summary: The applications of universal DNA base analogues. AB - A universal base analogue forms 'base pairs' with each of the natural DNA/RNA bases with little discrimination between them. A number of such analogues have been prepared and their applications as biochemical tools investigated. Most of these analogues are non-hydrogen bonding, hydrophobic, aromatic 'bases' which stabilise duplex DNA by stacking interactions. This review of the literature of universal bases (to 2000) details the analogues investigated, and their uses and limitations are discussed. PMID- 11410650 TI - The yeast POP2 gene encodes a nuclease involved in mRNA deadenylation. AB - The major mRNA degradation pathway involves deadenylation of the target molecule followed by decapping and, finally, 5'-->3' exonuclease digestion of the mRNA body. While yeast factors involved in the decapping and exonuclease degradation steps have been identified, the nature of the factor(s) involved in the deadenylation step remained elusive. Database searches for yeast proteins related to the mammalian deadenylase PARN identified the Pop2 protein (Pop2p) as a potential deadenylase. While Pop2p was previously identified as a factor affecting transcription, we identified a non-canonical RNase D sequence signature in its sequence. Analysis of the fate of a reporter mRNA in a pop2 mutant demonstrates that Pop2p is required for efficient mRNA degradation in vivo. Characterisation of mRNA degradation intermediates accumulating in this mutant supports the involvement of Pop2p in mRNA deadenylation in vivo. Similar phenotypes are observed in yeast strains lacking the Ccr4 protein, which is known to be associated with Pop2p. A recombinant Pop2p fragment encompassing the putative catalytic domain degrades poly(A) in vitro demonstrating that Pop2p is a nuclease. We also demonstrate that poly(A) is a better competitor than poly(G) or poly(C) of the Pop2p nuclease activity. Altogether, our study indicates that Pop2p is a nuclease subunit of the yeast deadenylase and suggests that Pop2p homologues in other species may have similar functions. PMID- 11410651 TI - Synthesis and properties of oligonucleotides containing 5-formyl-2' deoxycytidine: in vitro DNA polymerase reactions on DNA templates containing 5 formyl-2'-deoxycytidine. AB - Oligodeoxynucleotides (ODNs) containing 5-formyl-2'-deoxycytidine (fC) were synthesized by the phosphoramidite method and subsequent oxidation with sodium periodate. The stabilities of duplexes containing A, G, C or T opposite fC were studied by thermal denaturation. It was found that fC:A, fC:C or fC:T base pairs significantly reduce the thermal stabilities of duplexes. Next, single nucleotide insertion reactions were performed using ODNs containing fC as templates and the Klenow fragment of Escherichia coli DNA polymerase I. It was found that: (i) insertion of dGMP opposite fC appears to be less efficient relative to insertion opposite 5-methyl-2'-deoxycytidine (mC); (ii) dAMP is misincorporated more frequently opposite fC than mC, although the frequency of misincorporation seems to be dependent on the sequence; (iii) TMP is misincorporated more frequently opposite fC than mC. These results suggest that fC may induce the transition mutation C.G-->T.A and the transversion mutation C.G-->A.T during DNA synthesis. PMID- 11410652 TI - RecA-mediated strand exchange traverses substitutional heterologies more easily than deletions or insertions. AB - RecA protein in bacteria and its eukaryotic homolog Rad51 protein are responsible for initiation of strand exchange between homologous DNA molecules. This process is crucial for homologous recombination, the repair of certain types of DNA damage and for the reinitiation of DNA replication on collapsed replication forks. We show here, using two different types of in vitro assays, that in the absence of ATP hydrolysis RecA-mediated strand exchange traverses small substitutional heterologies between the interacting DNAs, whereas small deletions or insertions block the ongoing strand exchange. We discuss evolutionary implications of RecA selectivity against insertions and deletions and propose a molecular mechanism by which RecA can exert this selectivity. PMID- 11410653 TI - Non-independence of Mnt repressor-operator interaction determined by a new quantitative multiple fluorescence relative affinity (QuMFRA) assay. AB - Salmonella bacteriophage repressor Mnt belongs to the ribbon-helix-helix class of transcription factors. Previous SELEX results suggested that interactions of Mnt with positions 16 and 17 of the operator DNA are not independent. Using a newly developed high-throughput quantitative multiple fluorescence relative affinity (QuMFRA) assay, we directly quantified the relative equilibrium binding constants (K(ref)) of Mnt to operators carrying all the possible dinucleotide combinations at these two positions. Results show that Mnt prefers binding to C, instead of wild-type A, at position 16 when wild-type C at position 17 is changed to other bases. The measured K(ref) values of double mutants were also higher than the values predicted from single mutants, demonstrating the non-independence of these two positions. The ability to produce a large number of quantitative binding data simultaneously and the potential to scale up makes QuMFRA a valuable tool for the large-scale study of macromolecular interaction. PMID- 11410654 TI - The RNA-binding specificity of the mouse Dazl protein. AB - DAZ is an RNA-binding protein encoded by a region on the Y chromosome implicated in infertility, and DAZ-like (Dazl) proteins are master regulators of germ line gene expression in all animals. In mice Dazl is only expressed in germ cells and is necessary for meiosis. A dual approach was taken to understand the RNA-binding specificity of the Dazl protein: (i) traditional SELEX and (ii) a novel tri hybrid screen. Both approaches led to the same conclusion, namely that Dazl binds oligo(U) stretches interspersed by G or C residues. In a directed tri-hybrid assay the strongest interaction was with the consensus (GUn)n. This motif is found in the 5' UTR of CDC25C whose homologue is thought to be the target of Boule, the Dazl homologue in flies. CDC25C 5' UTR also interacted specifically with Dazl in vitro. The tri-hybrid screen retrieved UTRs of known genes that may be physiological substrates of Dazl. PMID- 11410655 TI - Gene expression in isolated plant mitochondria: high fidelity of transcription, splicing and editing of a transgene product in electroporated organelles. AB - Mitochondrial gene expression was studied using an electrotransformation protocol to introduce foreign DNA into purified wheat mitochondria. Optimal conditions for DNA uptake and transient gene expression were determined. We show here that a DNA plasmid containing either a cognate or a non-cognate gene under the control of a plant mitochondrial promoter is incorporated into the organelle and faithfully recognized by the transcription machinery. Transcripts generated by a plasmid bearing the intron-containing cox II gene were correctly spliced. Moreover, the transcripts were edited at the expected target C residues. The expression and maturation process of the transgene is dependent on the integrity of functional elements such as the promotor or the presence of structural domains necessary for splicing. The mitochondrial transformation described in this report is an important tool to study the multiple steps involved in plant mitochondrial gene expression at conditions closer to those found in vivo. PMID- 11410656 TI - The nicking endonuclease N.BstNBI is closely related to type IIs restriction endonucleases MlyI and PleI. AB - N.BstNBI is a nicking endonuclease that recognizes the sequence GAGTC and nicks the top strand preferentially. The Type IIs restriction endonucleases PleI and MlyI also recognize GAGTC, but cleave both DNA strands. Cloning and sequencing the genes encoding each of these three endonucleases discloses significant sequence similarities. Mutagenesis studies reveal a conserved set of catalytic residues among the three endonucleases, suggesting that they are closely related to each other. Furthermore, PleI and MlyI contain a single active site for DNA cleavage. The results from cleavage assays show that the reactions catalyzed by PleI and MlyI are sequential two step processes. The double-stranded DNA is first nicked on one DNA strand and then further cleaved on the second strand to form linear DNA. Gel filtration analysis shows that MlyI dimerizes in the presence of a cognate DNA and Ca(2+) whereas N.BstNBI remains a monomer, implicating dimerization as a requisite for the second strand cleavage. We suggest that N.BstNBI, MlyI and PleI diverged from a common ancestor and propose that N.BstNBI differs from MlyI and PleI in having an extremely limited second strand cleavage activity, resulting in a site-specific nicking endonuclease. PMID- 11410657 TI - An unusually compact external promoter for RNA polymerase III transcription of the human H1RNA gene. AB - H1 RNA, the RNA component of the human nuclear RNase P, is encoded by a unique gene transcribed by RNA polymerase III (Pol III). In this work, cis-acting elements and trans-acting factors involved in human H1 gene transcription were characterized by transcription assays of mutant templates and DNA binding assays of recombinant proteins. Four elements, lying within 100 bp of 5'-flanking sequences, were defined to be essential for maximal in vitro and in vivo expression, consisting of the octamer, Staf, proximal sequence element (PSE) and TATA motifs. These are also encountered in the promoter elements of vertebrate snRNA genes, where the first two constitute the distal sequence element (DSE). In all the genes examined so far, the DSE is distant from the PSE and TATA box that compose the basal promoter. However, we observed a fundamental difference in the organization of the H1 RNA and snRNA gene promoters with respect to the relative spacing of the DSE and PSE. Indeed, the H1 promoter is unusually compact, with the octamer motif and Staf binding site adjacent to the PSE and TATA motifs. It thus appears that the human RNase P RNA gene has adopted a unique promoter strategy placing the DSE immediately adjacent to the basal promoter. PMID- 11410658 TI - The participation of 5S rRNA in the co-translational formation of a eukaryotic 5S ribonucleoprotein complex. AB - The eukaryotic ribosomal 5S RNA-protein complex (5S rRNP) is formed by a co translational event that requires 5S rRNA binding to the nascent peptide chain of eukaryotic ribosomal protein L5. Binding between 5S rRNA and the nascent chain is specific: neither the 5S rRNA nor the nascent chain of L5 protein can be substituted by other RNAs or other ribosomal proteins. The region responsible for binding 5S rRNA is located at positions 35-50 with amino acid sequence RLVIQDIKNKYNTPKYRM. Eukaryotic 5S rRNA binds a nascent chain having this sequence, but such binding is not substantive enough to form a 5S-associated RNP complex, suggesting that 5S rRNA binding to the nascent chain is amino acid sequence dependent and that formation of the 5S rRNP complex is L5 protein specific. Microinjection of 5S rRNP complex into the cytoplasm of Xenopus oocytes results in both an increase in the initial rate and also in the extent of net nuclear import of L5. This suggests that the 5S rRNP complex enhances nuclear transport of L5. We propose that 5S rRNA plays a chaperone-like role in folding of the nascent chain of L5 and directs L5 into a 5S rRNP complex for nuclear entry. PMID- 11410659 TI - Mi2, an auto-antigen for dermatomyositis, is an ATP-dependent nucleosome remodeling factor. AB - Dynamic changes in chromatin structure play an important role in transcription regulation. Recent studies have revealed two mechanisms that alter chromatin structure. One involves ATP-dependent chromatin remodeling, and the other involves acetylation of the core histone tails. We have previously purified and characterized a multi-subunit protein complex, NuRD, which possesses both nucleosome remodeling and histone deacetylase activities. Despite extensive biochemical characterization of the complex, little is known about the functions of its individual components. In this study, we focused on Mi2, a component of the NuRD complex. We found that, similar to the native NuRD complex, recombinant Mi2 is a DNA-dependent, nucleosome-stimulated ATPase. Kinetic analysis of the ATP hydrolysis reaction indicated that the differential stimulation of the Mi2 ATPase by DNA and nucleosomes were primarily due to their differential effects on the turnover number of the reaction. Furthermore, we demonstrated that recombinant Mi2 is an efficient nucleosome remodeling factor when compared to that of the native NuRD complex. Our results define the biochemical function of Mi2 and set the stage for understanding the mechanism of nucleosome remodeling in a defined reconstituted system. PMID- 11410660 TI - The effect of amino groups on the stability of DNA duplexes and triplexes based on purines derived from inosine. AB - The effect of amino groups attached at positions 2 and 8 of the hypoxanthine moiety in the structure, reactivity and stability of DNA duplexes and triplexes is studied by means of quantum mechanical calculations, as well as extended molecular dynamics (MD) and thermodynamic integration (MD/TI) simulations. Theoretical estimates of the change in stability related to 2'-deoxyguanosine (G) --> 2'-deoxyinosine (I) --> 8-amino-2'-deoxyinosine (8AI) mutations have been experimentally verified, after synthesis of the corresponding compounds. An amino group placed at position 2 stabilizes the duplex, as expected, and surprisingly also the triplex. The presence of an amino group at position 8 of the hypoxanthine moiety stabilizes the triplex but, surprisingly, destabilizes the duplex. The subtle electronic redistribution occurring upon the introduction of an amino group on the purine seems to be responsible for this surprising behavior. Interesting 'universal base' properties are found for 8AI. PMID- 11410661 TI - Hepatitis C virus internal ribosome entry site RNA contains a tertiary structural element in a functional domain of stem-loop II. AB - The internal ribosome entry site (IRES) of hepatitis C virus (HCV) RNA contains >300 bases of highly conserved 5'-terminal sequence, most of it in the uncapped 5'-untranslated region (5'-UTR) upstream from the single AUG initiator triplet at which translation of the HCV polyprotein begins. Although progress has been made in defining singularities like the RNA pseudoknot near this AUG, the sequence and structural features of the HCV IRES which stimulate accurate and efficient initiation of protein synthesis are only partially defined. Here we report that a region further upstream from the AUG, stem-loop II of the HCV IRES, also contains an element of local tertiary structure which we have detected using RNase H cleavage and have mapped using the singular ability of two bases therein to undergo covalent intra-chain crosslinking stimulated by UV light. This pre existing element maps to two non-contiguous stretches of the HCV IRES sequence, residues 53-68 and 103-117. Several earlier studies have shown that the correct sequence between bases 45 and 70 of the HCV IRES stem-loop II domain is required for initiation of protein synthesis. Because features of local tertiary structure like the one we report here are often associated with protein binding, we propose that the HCV stem-loop II element is directly involved in IRES action. PMID- 11410662 TI - A novel cell-free system reveals a mechanism of circular DNA formation from tandem repeats. AB - One characteristic of genomic plasticity is the presence of extrachromosomal circular DNA (eccDNA). High levels of eccDNA are associated with genomic instability, exposure to carcinogens and aging. We have recently reported developmentally regulated formation of eccDNA that occurs preferentially in pre blastula Xenopus laevis embryos. Multimers of tandemly repeated sequences were over-represented in the circle population while dispersed sequences were not detected, indicating that circles were not formed at random from any chromosomal sequence. Here we present detailed mechanistic studies of eccDNA formation in a cell-free system derived from Xenopus egg extracts. We show that naked chromosomal DNA from sperm or somatic tissues serves as a substrate for direct tandem repeat circle formation. Moreover, a recombinant bacterial tandem repeat can generate eccDNA in the extract through a de novo mechanism which is independent of DNA replication. These data suggest that the presence of a high level of any direct tandem repeat can confer on DNA the ability to be converted into circular multimers in the early embryo irrespective of its sequence and that homologous recombination is involved in this process. PMID- 11410663 TI - Issues in cDNA microarray analysis: quality filtering, channel normalization, models of variations and assessment of gene effects. AB - We consider the problem of comparing the gene expression levels of cells grown under two different conditions using cDNA microarray data. We use a quality index, computed from duplicate spots on the same slide, to filter out outlying spots, poor quality genes and problematical slides. We also perform calibration experiments to show that normalization between fluorescent labels is needed and that the normalization is slide dependent and non-linear. A rank invariant method is suggested to select non-differentially expressed genes and to construct normalization curves in comparative experiments. After normalization the residuals from the calibration data are used to provide prior information on variance components in the analysis of comparative experiments. Based on a hierarchical model that incorporates several levels of variations, a method for assessing the significance of gene effects in comparative experiments is presented. The analysis is demonstrated via two groups of experiments with 125 and 4129 genes, respectively, in Escherichia coli grown in glucose and acetate. PMID- 11410664 TI - A novel fluorometric oligonucleotide assay to measure O( 6)-methylguanine DNA methyltransferase, methylpurine DNA glycosylase, 8-oxoguanine DNA glycosylase and abasic endonuclease activities: DNA repair status in human breast carcinoma cells overexpressing methylpurine DNA glycosylase. AB - DNA repair status plays a major role in mutagenesis, carcinogenesis and resistance to genotoxic agents. Because DNA repair processes involve multiple enzymatic steps, understanding cellular DNA repair status has required several assay procedures. We have developed a novel in vitro assay that allows quantitative measurement of alkylation repair via O(6)-methylguanine DNA methyltransferase (MGMT) and base excision repair (BER) involving methylpurine DNA glycosylase (MPG), human 8-oxoguanine DNA glycosylase (hOGG1) and yeast and human abasic endonuclease (APN1 and APE/ref-1, respectively) from a single cell extract. This approach involves preparation of cell extracts in a common buffer in which all of the DNA repair proteins are active and the use of fluorometrically labeled oligonucleotide substrates containing DNA lesions specific to each repair protein. This method enables methylation and BER capacities to be determined rapidly from a small amount of starting sample. In addition, the stability of the fluorometric oligonucleotides precludes the substrate variability caused by continual radiolabeling. In this report this technique was applied to human breast carcinoma MDA-MB231 cells overexpressing human MPG in order to assess whether up-regulation of the initial step in BER alters the activity of selected other BER (hOGG1 and APE/ref-1) or direct reversal (MGMT) repair activities. PMID- 11410665 TI - The brefeldin A resistance protein Bfr1p is a component of polyribosome associated mRNP complexes in yeast. AB - The yeast gene BFR1 was originally isolated from a genetic screen for high-copy suppressors of brefeldin A-induced lethality in Saccharomyces cerevisiae. While this result suggested a possible role for the encoded protein, Bfr1p, in the secretory pathway, subsequent data have not fully supported this conclusion. Alternatively, Bfr1p has also been found by yeast two-hybrid analysis to interact with Bbp1p, a component of the spindle pole body. Finally, we have reported that Bfr1p associates with cytoplasmic mRNP complexes containing Scp160p, raising the possibility that Bfr1p may function in mRNA metabolism. Here, we have explored this possibility further. We report that Bfr1p associates with yeast polyribosomes and mRNP complexes even in the absence of Scp160p, and that its interaction with Scp160p-containing mRNP complexes is RNA-dependent. Furthermore, we have determined by fluorescence microscopy and subcellular fractionation that Bfr1p and Scp160p demonstrate similar cytoplasmic localization with enrichment around the nuclear envelope/endoplasmic reticulum. Finally, we report that loss of Bfr1p disrupts the interaction of Scp160p with polyribosomes, thereby demonstrating that the relationship between these two proteins is functional as well as physical. Considered together, these data raise the intriguing possibility that Bfr1p may provide a link between mRNA metabolism, the chromosomal segregation machinery and perhaps secretion in yeast. PMID- 11410666 TI - A mRNA determinant of gRNA-directed kinetoplastid editing. AB - Several mitochondrial mRNAs of the kinetoplastid protozoa do not encode a functional open reading frame until they have been edited through the addition or deletion of U nucleotides at specific sites. Genetic information specifying the location and extent of editing is present on guide RNAs (gRNAs). The sequence adjacent to most mRNA editing sites has a high purine content which previously has been proposed to facilitate the editing reaction through base-pairing to a poly(U) tail at the 3' end of the gRNA. We demonstrate here that gRNA binding alone is insufficient to create an editing site and that the mRNA sequence near an editing site is an additional determinant affecting the efficiency of the reaction. PMID- 11410667 TI - Human GC-AG alternative intron isoforms with weak donor sites show enhanced consensus at acceptor exon positions. AB - It has been previously observed that the intrinsically weak variant GC donor sites, in order to be recognized by the U2-type spliceosome, possess strong consensus sequences maximized for base pair formation with U1 and U5/U6 snRNAs. However, variability in signal strength is a fundamental mechanism for splice site selection in alternative splicing. Here we report human alternative GC-AG introns (for the first time from any species), and show that while constitutive GC-AG introns do possess strong signals at their donor sites, a large subset of alternative GC-AG introns possess weak consensus sequences at their donor sites. Surprisingly, this subset of alternative isoforms shows strong consensus at acceptor exon positions 1 and 2. The improved consensus at the acceptor exon can facilitate a strong interaction with U5 snRNA, which tethers the two exons for ligation during the second step of splicing. Further, these isoforms nearly always possess alternative acceptor sites and exhibit particularly weak polypyrimidine tracts characteristic of AG-dependent introns. The acceptor exon nucleotides are part of the consensus required for the U2AF(35)-mediated recognition of AG in such introns. Such improved consensus at acceptor exons is not found in either normal or alternative GT-AG introns having weak donor sites or weak polypyrimidine tracts. The changes probably reflect mechanisms that allow GC-AG alternative intron isoforms to cope with two conflicting requirements, namely an apparent need for differential splice strength to direct the choice of alternative sites and a need for improved donor signals to compensate for the central mismatch base pair (C-A) in the RNA duplex of U1 snRNA and the pre-mRNA. The other important findings include (i) one in every twenty alternative introns is a GC-AG intron, and (ii) three of every five observed GC-AG introns are alternative isoforms. PMID- 11410668 TI - Mutations in the TAR hairpin affect the equilibrium between alternative conformations of the HIV-1 leader RNA. AB - The HIV-1 untranslated leader RNA can adopt two mutually exclusive conformations that represent alternative secondary structures. This leader RNA can fold either an extended duplex through long-distance base pairing or a branched conformation in which the RNA locally folds into hairpin structures. Both leader RNA conformations have the TAR hairpin in common, which forms the extreme 5' end of all HIV-1 transcripts. We report that truncation of the TAR hairpin shifts the equilibrium between the two RNA conformations away from the thermodynamically favored long-distance interaction. However, the equilibrium is partially restored in response to the cations Na(+) and Mg(2+). The transcripts with mutant TAR structures allowed us to investigate conditions affecting the competition between the alternative conformations of the HIV-1 leader RNA. We also demonstrate that the change in conformation of the leader RNA due to TAR truncations severely affects formation of the HIV-1 RNA dimer. PMID- 11410669 TI - Structure in nascent RNA leads to termination of slippage transcription by T7 RNA polymerase. AB - T7 RNA polymerase presents a very simple model system for the study of fundamental aspects of transcription. Some time ago it was observed that in the presence of only GTP as a substrate, on a template encoding the initial sequence GGGA., T7 RNA polymerase will synthesize a 'ladder' of poly-G RNA products. At each step, the ratio of elongation to product release is consistently approximately 0.75 until the RNA reaches a length of approximately 13-14 nt, at which point this ratio drops precipitously. One model to explain this drop in complex stability suggests that the nascent RNA may be structurally hindered by the protein; the RNA may be exiting via a pathway not taken by normally synthesized RNA and therefore becomes sterically destabilized. The fact that the length of RNA at which this occurs is close to the length at which the transition to a stably elongating complex occurs might have led to other mechanistic proposals. Here we show instead that elongation falls off due to the cooperative formation of structure in the nascent RNA, most likely an intramolecular G quartet structure. Replacement of GTP by 7-deaza-GTP completely abolishes this transition and G-ladder synthesis continues with a constant efficiency of elongation beyond the limit of detection. The polymerase-DNA complex creates no barrier to the growth of the nascent (slippage) RNA, rather termination is similar to that which occurs in rho-independent termination. PMID- 11410670 TI - GeneMarkS: a self-training method for prediction of gene starts in microbial genomes. Implications for finding sequence motifs in regulatory regions. AB - Improving the accuracy of prediction of gene starts is one of a few remaining open problems in computer prediction of prokaryotic genes. Its difficulty is caused by the absence of relatively strong sequence patterns identifying true translation initiation sites. In the current paper we show that the accuracy of gene start prediction can be improved by combining models of protein-coding and non-coding regions and models of regulatory sites near gene start within an iterative Hidden Markov model based algorithm. The new gene prediction method, called GeneMarkS, utilizes a non-supervised training procedure and can be used for a newly sequenced prokaryotic genome with no prior knowledge of any protein or rRNA genes. The GeneMarkS implementation uses an improved version of the gene finding program GeneMark.hmm, heuristic Markov models of coding and non-coding regions and the Gibbs sampling multiple alignment program. GeneMarkS predicted precisely 83.2% of the translation starts of GenBank annotated Bacillus subtilis genes and 94.4% of translation starts in an experimentally validated set of Escherichia coli genes. We have also observed that GeneMarkS detects prokaryotic genes, in terms of identifying open reading frames containing real genes, with an accuracy matching the level of the best currently used gene detection methods. Accurate translation start prediction, in addition to the refinement of protein sequence N-terminal data, provides the benefit of precise positioning of the sequence region situated upstream to a gene start. Therefore, sequence motifs related to transcription and translation regulatory sites can be revealed and analyzed with higher precision. These motifs were shown to possess a significant variability, the functional and evolutionary connections of which are discussed. PMID- 11410671 TI - Experimental evaluation of the Liu-Beveridge dinucleotide step model of DNA structure. AB - Methods for predicting DNA curvature have many possible applications. Dinucleotide step models describe DNA shape by characterization of helical twist, deflection angles and the direction of deflection for nearest neighbor base pairs. Liu and Beveridge have extended previous applications of dinucleotide step models with the development and qualitative validation of a predictive method for sequence-dependent DNA curvature (the LB model). We tested whether the LB model accurately predicts experimentally deduced curvature angles and helical repeat parameters for DNA sequences not in its training set, particularly when challenged with quantitative data and subtle sequence phasings. We examined a series of 17 well-characterized DNA sequences to compare electrophoretic and computational results. The LB model is superior to two other models in the prediction of helical repeat parameters. We observed a strong linear correlation between curvature magnitudes predicted using the LB model and those determined by electrophoretic ligation ladder experiments, although the LB model somewhat underestimated apparent curvature. With longer electrophoretic phasing probes the LB model slightly overestimated gel mobility anomalies, with modest deviations in predicted helical repeat parameters. Overall, our analyses suggest that the LB model provides reasonably accurate predictions for the electrophoretic behavior of DNA. PMID- 11410672 TI - Binding specificity and stability of duplexes formed by modified oligonucleotides with a 4096-hexanucleotide microarray. AB - The binding of oligodeoxynucleotides modified with adenine 2'-O-methyl riboside, 2,6-diaminopurine 2'-O-methyl riboside, cytosine 2'-O-methyl riboside, 2,6 diaminopurine deoxyriboside or 5-bromodeoxyuridine was studied with a microarray containing all possible (4096) polyacrylamide-bound hexadeoxynucleotides (a generic microchip). The generic microchip was manufactured by using reductive immobilization of aminooligonucleotides in the activated copolymer of acrylamide, bis-acrylamide and N-(2,2-dimethoxyethyl) acrylamide. The binding of the fluorescently labeled modified octanucleotides to the array was analyzed with the use of both melting profiles and the fluorescence distribution at selected temperatures. Up to three substitutions of adenosines in the octamer sequence by adenine 2'-O-methyl ribosides (A(m)), 2,6-diaminopurine 2'-O-methyl ribosides (D(m)) or 2,6-diaminopurine deoxyribosides (D) resulted in increased mismatch discrimination measured at the melting temperature of the corresponding perfect duplex. The stability of complexes formed by 2'-O-methyl-adenosine-modified oligodeoxynucleotides was slightly decreased with every additional substitution, yielding approximately 4 degrees C of total loss in melting temperature for three modifications, as followed from microchip thermal denaturation experiments. 2,6 Diaminopurine 2'-O-methyl riboside modifications led to considerable duplex stabilization. The cytosine 2'-O-methyl riboside and 5-bromodeoxyuridine modifications generally did not change either duplex stability or mismatch resolution. Denaturation experiments conducted with selected perfect duplexes on microchips and in solution showed similar results on thermal stabilities. Some hybridization artifacts were observed that might indicate the formation of parallel DNA. PMID- 11410673 TI - DNA-XPA interactions: a (31)P NMR and molecular modeling study of dCCAATAACC association with the minimal DNA-binding domain (M98-F219) of the nucleotide excision repair protein XPA. AB - Recent NMR-based, chemical shift mapping experiments with the minimal DNA-binding domain of XPA (XPA-MBD: M98-F219) suggest that a basic cleft located in the loop rich subdomain plays a role in DNA-binding. Here, XPA-DNA interactions are further characterized by NMR spectroscopy from the vantage point of the DNA using a single-stranded DNA nonamer, dCCAATAACC (d9). Up to 2.5 molar equivalents of XPA-MBD was titrated into a solution of d9. A subset of (31)P resonances of d9 were observed to broaden and/or shift providing direct evidence that XPA-MBD binds d9 by a mechanism that perturbs the phosphodiester backbone of d9. The interior five residues of d9 broadened and/or shifted before (31)P resonances of phosphate groups at the termini, suggesting that when d9 is bound to XPA-MBD the internal residues assume a correlation time that is characteristic of the molecular weight of the complex while the residues at the termini undergo a fraying motion away from the surface of the protein on a timescale such that the line widths are more characteristic of the molecular weight of ssDNA. A molecular model of the XPA-MBD complex with d9 was calculated based on the (15)N (XPA-MBD) and (31)P (d9) chemical shift mapping studies and on the assumption that electrostatic interactions drive the complex formation. The model shows that a nine residue DNA oligomer fully covers the DNA-binding surface of XPA and that there may be an energetic advantage to binding DNA in the 3'-->5' direction rather than in the 5'-->3' direction (relative to XPA-MBD alpha-helix-3). PMID- 11410674 TI - Folding of the group I intron ribozyme from the 26S rRNA gene of Candida albicans. AB - Preincubation of the group I intron Ca.LSU from Candida albicans at 37 degrees C in the absence of divalent cations results in partial folding of this intron. This is indicated by increased resistance to T1 ribonuclease cleavage of many G residues in most local helices, including P4-P6, as well as the non-local helix P7, where the G binding site is located. These changes correlate with increased gel mobility and activation of catalysis by precursor RNA containing this intron after preincubation. The presence of divalent cations or spermidine during preincubation results in formation of the predicted helices, as indicated by protection of additional G residues. However, addition of these cations during preincubation of the precursor RNA alters its gel mobility and eliminates the preincubation activation of precursor RNA seen in the absence of cations. These results suggest that, in the presence of divalent cations or spermidine, Ca.LSU folds into a more ordered, stable but misfolded conformation that is less able to convert into the catalytically active form than the ribozyme preincubated without cations. These results indicate that, like the group I intron of Tetrahymena, multiple folding pathways exist for Ca.LSU. However, it appears that the role cations play in the multiple folding pathways leading to the catalytically active form may differ between folding of these two group I introns. PMID- 11410675 TI - Massive parallel analysis of the binding specificity of histone-like protein HU to single- and double-stranded DNA with generic oligodeoxyribonucleotide microchips. AB - A generic hexadeoxyribonucleotide microchip has been applied to test the DNA binding properties of HU histone-like bacterial protein, which is known to have a low sequence specificity. All 4096 hexamers flanked within 8mers by degenerate bases at both the 3'- and 5'-ends were immobilized within the 100 x 100 x 20 mm polyacrylamide gel pads of the microchip. Single-stranded immobilized oligonucleotides were converted in some experiments to the double-stranded form by hybridization with a specified mixture of 8mers. The DNA interaction with HU was characterized by three type of measurements: (i) binding of FITC-labeled HU to microchip oligonucleotides; (ii) melting curves of complexes of labeled HU with single-stranded microchip oligonucleotides; (iii) the effect of HU binding on melting curves of microchip double-stranded DNA labeled with another fluorescent dye, Texas Red. Large numbers of measurements of these parameters were carried out in parallel for all or many generic microchip elements in real time with a multi-wavelength fluorescence microscope. Statistical analysis of these data suggests some preference for HU binding to G/C-rich single-stranded oligonucleotides. HU complexes with double-stranded microchip 8mers can be divided into two groups in which HU binding either increased the melting temperature (T(m)) of duplexes or decreased it. The stabilized duplexes showed some preference for presence of the sequence motifs AAG, AGA and AAGA. In the second type of complex, enriched with A/T base pairs, the destabilization effect was higher for longer stretches of A/T duplexes. Binding of HU to labeled duplexes in the second type of complex caused some decrease in fluorescence. This decrease also correlates with the higher A/T content and lower T(m). The results demonstrate that generic microchips could be an efficient approach in analysis of sequence specificity of proteins. PMID- 11410676 TI - The first phytoplasma RNase P RNA provides new insights into the sequence requirements of this ribozyme. AB - A high variability of RNase P RNA structures is seen among members of the Mycoplasma group. To gain further insight into the structure-function relations of this ribozyme, we have searched for the RNase P RNA gene from more distant relatives, the phytoplasmas. These mycoplasma-like organisms are the aetiological agents of many severe plant diseases. We report the sequence and catalytic properties of RNase P RNA from the phytoplasma causing apple proliferation disease. The primary and postulated secondary structure of this 443 nt long RNA are most similar to those of Acholeplasma, supporting the phylogenetic position of this pathogen. Remarkably, the extremely AT-rich (73.6%) phytoplasma RNA differs from the known bacterial consensus sequence by a single base pair, which is positioned close to the substrate cleavage site in current three-dimensional models. Phytoplasma RNase P RNA functions as an efficient ribozyme in vitro. Conversion of its sequence to the full consensus and kinetic analysis of the resulting mutant RNAs suggests that neither the sequence alone, nor the type of pairing at this position is crucial for substrate binding or catalysis by the RNase P ribozyme. These results refine the bacterial consensus structure close to the catalytic core and thus improve our understanding of RNase P RNA function. PMID- 11410677 TI - Differential DNA recognition and glycosylase activity of the native human MutY homolog (hMYH) and recombinant hMYH expressed in bacteria. AB - Human MutY homolog (hMYH), an adenine DNA glycosylase, can effectively remove misincorporated adenines opposite template G or 8-oxoG bases, thereby preventing G:C-->T:A transversions. Human cell extracts possess the adenine DNA glycosylase activity of hMYH and can form protein-DNA complexes with both A/G and A/8-oxoG mismatches. hMYH in cell extracts was shown to be the primary binding protein for A/G- and A/8-oxoG-containing DNA substrates by UV cross-linking. However, recombinant hMYH expressed in bacteria has much weaker glycosylase and substrate binding activities towards A/G mismatches than native hMYH. Moreover, the protein DNA complex of bacterially expressed hMYH migrates much faster than that of native hMYH in a non-denaturing polyacrylamide gel. Dephosphorylation of native hMYH reduces the glycosylase activity on A/G more extensively than on A/8-oxoG mismatches but does not alter the gel mobility of the protein-DNA complex. Our results suggest that hMYH in human cell extracts may be associated with other factors in the protein-DNA complex to account for its slower mobility in the gel. hMYH and apurinic/apyrimidinic endonuclease (hAPE1) co-migrate with the protein DNA complex formed by the extracts and A/8-oxoG-containing DNA. PMID- 11410678 TI - Efficient and heritable functional knock-out of an adult phenotype in Drosophila using a GAL4-driven hairpin RNA incorporating a heterologous spacer. AB - We have developed a modified RNA interference (RNAi) method for generating gene knock-outs in Drosophila melanogaster. We used the sequence of the yellow (y) locus to construct an inverted repeat that will form a double-stranded hairpin structure (y-IR) that is under the control of the upstream activating sequence (UAS) of the yeast transcriptional activator GAL4. Hairpins are extremely difficult to manipulate in Escherichia coli, so our method makes use of a heterologous 330 bp spacer encoding sequences from green fluorescent protein to facilitate the cloning steps. When the UAS-y-IR hairpin is expressed under the control of different promoter-GAL4 fusions, a high frequency of y pigment phenocopies is obtained in adults. Consequently this method for producing gene knock-outs has several advantages over previous methods in that it is applicable to any gene within the fly genome, greatly facilitates cloning of the hairpin, can be used if required with GAL4 drivers to avoid lethality or to induce RNAi in a specific developmental stage and/or tissue, is useful for generating knock-outs of adult phenotypes as reported here and, finally, the system can be manipulated to investigate the trans-acting factors that are involved in the RNAi mechanism. PMID- 11410679 TI - A single vector containing modified cre recombinase and LOX recombination sequences for inducible tissue-specific amplification of gene expression. AB - The selective alteration of the genome using Cre recombinase to target the rearrangement of genes flanked by LOX recognition sequences has required the use of two separate genetic constructs in trans, one containing cre and the other containing the gene of interest flanked by LOX sites. We have developed a strategy in which both the cre recombinase gene and LOX recombination sites may be cloned within a single vector in cis. This method uses a modified form of Cre (CREM) that contains alterations to the 5' region including the introduction of a Kozak consensus sequence and insertion of a functional intron. This system allows for the inducible, tissue-specific activation or inactivation of gene expression in a single vector and can be utilized for the 300-fold amplification of gene expression from a weak promoter. This approach can be applied to targeting strategies for generating genetically altered mice and gene therapy. PMID- 11410680 TI - A new G-tailing method for the determination of the poly(A) tail length applied to hepatitis A virus RNA. AB - To study the role of the poly(A) tail length during the replication of poly(A) containing plus-strand RNA virus, we have developed a simple reverse transcription polymerase chain reaction (RT-PCR)-based method that substantially improves the previously reported PAT [poly(A) test] assay. In contrast to the PAT assay, the new method is based on the enzymatic 3' elongation of mRNA with guanosine residues, thus immediately preserving the 3' end of the RNA and creating a unique poly(A)-oligo(G) junction. The oligo(G)-protected full-length poly(A) tail is reverse transcribed using the universal anti-sense primer oligo(dC(9)T(6)) and amplified by PCR with a gene-specific sense primer. After sequencing the resulting RT-PCR product the length of the poly(A) tail was unequivocally deduced from the number of adenosine residues between the oligo(G) stretch and the sequence upstream of the poly(A) tail. The efficiency and specificity of the newly developed assay was demonstrated by analysing the poly(A) tail length of the hepatitis A virus (HAV) RNA. We show here that the poly(A) tail of HAV RNA rescued after transfection of in vitro transcripts was elongated in the course of HAV replication. PMID- 11410681 TI - Real-time detection of gene promoter activity: quantitation of toxin gene transcription. AB - We have developed a new method for quantification of promoter activity in cell lines transfected with recombinant plasmids containing the reporter gene encoding chloramphenicol acetyl transferase (CAT) by real-time PCR. As the efficiency of transfection has a direct influence on the total mRNA produced, we have used the neomycin-resistance gene present within the same vector DNA to normalize the measurement of mRNA levels. Three promoters from genes encoding toxins (pre synaptic neurotoxin phospholipase A(2), post-synaptic alpha neurotoxin and cardiotoxin), believed to have evolved from the same ancestor but exhibiting different promoter activities, have been employed in this study to demonstrate the feasibility and accuracy of the method in CAT gene reporter analysis. PMID- 11410682 TI - Yeast vectors for integration at the HO locus. AB - We have constructed new yeast vectors for targeted integration of desired sequences at the Saccharomyces cerevisiae HO locus. Insertion at HO has been shown to have no effect on yeast growth, and thus these integrations should be neutral. One vector contains the KanMX selectable marker, and integrants can be selected by resistance to G418. The other vector contains the hisG-URA3-hisG cassette, and integrants can be selected by uracil prototrophy. Subsequent growth on 5-FOA permits identification of colonies where recombination between the hisG tandem repeats has led to loss of the URA3 marker and return to uracil auxotrophy. We also describe several new bacterial polylinker vectors derived from pUC21 (ampicillin resistance) and pUK21 (kanamycin resistance). PMID- 11410683 TI - Identification and prevention of a GC content bias in SAGE libraries. AB - Serial Analysis of Gene Expression (SAGE) is becoming a widely used gene expression profiling method for the study of development, cancer and other human diseases. Investigators using SAGE rely heavily on the quantitative aspect of this method for cataloging gene expression and comparing multiple SAGE libraries. We have developed additional computational and statistical tools to assess the quality and reproducibility of a SAGE library. Using these methods, a critical variable in the SAGE protocol was identified that has the potential to bias the Tag distribution relative to the GC content of the 10 bp SAGE Tag DNA sequence. We also detected this bias in a number of publicly available SAGE libraries. It is important to note that the GC content bias went undetected by quality control procedures in the current SAGE protocol and was only identified with the use of these statistical analyses on as few as 750 SAGE Tags. In addition to keeping any solution of free DiTags on ice, an analysis of the GC content should be performed before sequencing large numbers of SAGE Tags to be confident that SAGE libraries are free from experimental bias. PMID- 11410684 TI - Carbon monoxide poisoning: systemic manifestations and complications. AB - Carbon monoxide (CO) has the toxic effects of tissue hypoxia and produces various systemic and neurological complications. The main clinical manifestations of acute CO poisoning consist of symptoms caused by alterations of the cardiovascular system such as initial tachycardia and hypertension, and central nervous system symptoms such as headache, dizziness, paresis, convulsion and unconsciousness. CO poisoning also produces myocardial ischemia, atrial fibrillation, pneumonia, pulmonary edema, erythrocytosis, leucocytosis, hyperglycemia, muscle necrosis, acute renal failure, skin lesion, and changes in perception of the visual and auditory systems. Of considerable clinical interest, severe neurological manifestations may occur days or weeks after acute CO poisoning. Delayed sequelae of CO poisoning are not rare, usually occur in middle or older, and are clinically characterized by symptom triad of mental deterioration, urinary incontinence, and gait disturbance. Occasionally, movement disorders, particularly parkinsonism, are observed. In addition, peripheral neuropathy following CO poisoning usually occurs in young adults. PMID- 11410685 TI - Molecular screening for fragile X syndrome in mentally handicapped children in Korea. AB - Fragile X syndrome is one of the most common forms of inherited mental retardation and is caused by the expansion of the CGG trinucleotide repeats in the FMR-1 gene. This study was aimed to facilitate the molecular screening of fragile X syndrome in Korean children with mental retardation of unknown etiology. The subjects were tested by Expand Long Template PCR system in the presence of 7-deaza-dGTP, and then by Southern blot analysis. The PCR method provided rapid and reliable results for the identification of fragile X negative and positive patients. One hundred one mentally retarded children (78 males and 23 females) were screened by PCR amplification, which detected only one abnormal sample. The PCR-positive case was confirmed by the CGG repeat expansion on Southern blot analysis with a positive cytogenetic result. In conclusion, Expand Long Template PCR may be used as the first screening test for detecting the fragile X syndrome. PMID- 11410686 TI - Clinical application of histamine prick test for food challenge in atopic dermatitis. AB - Determining positive food challenges are not easy as there is an absence of simple and objective tests. Histamine, an essential mediator for allergic reactions, is involved in the pathogenesis of atopic dermatitis (AD) and food challenges can change histamine levels. The significances of a prick test with histamine (histamine prick test, HPT) relating to the interpretation of food challenges in AD were evaluated. A total of 467 AD patients participated in this study. Skin prick tests, identification of specific IgE and open food challenge were conducted for the identification of food allergy. Elimination diet was performed with HPT. HPTs were conducted before and after food challenges. The wheal sizes by HPT were significantly decreased after an elimination diet. The relative changes of wheal sizes significantly correlated with those of clinical severity scores in AD patients (p<0.001). The wheal sizes in HPT were increased with a positive provocation in open food challenges. In conclusion, HPT may be a simple and objective test to interprete the results of food challenges in patients with AD. The exact mechanisms of the changes in skin reactivity by HPT need further investigation. PMID- 11410687 TI - Development of semi-empirical model for radiochemical reactions. AB - A mathematical model, which represents the radio-chemical reactions in water, was developed to study the effect of the radio-chemical products on cell killing. The five differential equations were solved using dose rate equation and cell survival as a function of dose was computed. The known chemical rate constants were taken from the literature and unknown constants were determined by curve fitting to an experimental data. Sensitivity studies were performed by varying the rate constants and showed that the yield of H-radical had little effect whereas the change in concentration of OH-radical and direct interaction resulted in significant change on cell survival. The sensitivity studies showed good agreement with the observed effects. In conclusion, we developed a mathematical model that could be used as a means for the estimation of radiation damage. PMID- 11410688 TI - Frequencies of fetal chromosomal abnormalities at prenatal diagnosis: 10 years experiences in a single institution. AB - We present frequencies of fetal chromosomal abnormalities in 4,907 prenatal cytogenetic examinations at Samsung Cheil Hospital from 1988 to 1997 for 10 yr duration. Prenatal karyotypes were undertaken in 3,913 amniotic fluid samples, 800 chorionic villi samples, and 194 percutaneous umbilical blood samples. The frequency of fetal abnormal karyotypes was 3.1% (150 cases). Numerical chromosome abnormalities were 87 cases (1.8%) and structural aberrations of chromosomes were 63 cases (1.3%). In the numerical chromosomal abnormalities, the frequency of trisomy 21 was by far the highest (36 cases), followed by trisomy 18 in 22 cases and sex chromosome aneuploidies in 19 cases. In the structural chromosomal aberrations, 5 cases had the inversions in chromosome 2, 7, 17, and Y. Chromosomal deletions in 6 cases and additions in 4 cases were analysed. Of the remaining 47 translocation in abnormal fetuses, reciprocal translocation was in 26 cases and Robertsonian translocation in 21 cases. Among them, 41 cases were balanced translocation and 6 were unbalanced. Thirty five cases of translocation were inherited from one of the parents. Four had de novo chromosome rearrangements, and 8 cases were unknown. PMID- 11410689 TI - Viable myocardium in reperfused acute myocardial infarction: rest and stress first-pass mr imaging. AB - Feasibility of identifying viable myocardium in rest and stress magnetic resonance imaging (MRI) was evaluated using 3 hr occlusion and 30 min reperfusion model of left anterior descending (LAD) coronary artery in 12 felines. At rest MRI, viable myocardium confirmed by 2,3,5-triphenyl tetrazolium chloride (TTC) staining showed rapid signal intensity (SI) rise followed by gradual decline not significantly different from normal myocardium that the two hyperperfused regions were distinguishable only from the hypoperfused nonviable myocardium. At stress MRI, hyperemia induced perfusion change was most pronounced in normal myocardium with earlier and greater peak enhancement followed by brisk 'washout' phase while minimally augmented enhancement in viable myocardium was still in 'washin' phase. From these findings, it was concluded that viable myocardium is identified in rest and stress MRI as redistributing hypo- perfusion compared to persistent hyper-perfusion of the normal myocardium and the persistent hypo-perfusion of the nonviable myocardium. PMID- 11410690 TI - Anastomosis of vessels less than 2 mm with the vascular clip system clip applier. AB - Sutures may cause endothelial trauma and occlusion. The vascular clip system (VCS) clip applier may minimize endothelial injury. Fourteen carotid arteries of nine adult rabbits were transected and re-anastomosed with either #7-0 polypropylene (Group I, n=8) or VCS clips (Group II, n=6). The animals were sacrificed at 1, 3, 8, 14, and 30 days postoperatively. The operation time and bleeding amount were checked for each anastomosis. Carotid angiograms, photography, H&E staining and scanning electron microscopy (SEM) were performed. Fibrin and thrombus, inflammatory cell infiltration, endothelial disruption, luminal distortion, fibrosis, and wall thickening were compared. The luminal diameter was greater in group II. There were minimal differences in thrombosis, wall thickening and fibrosis between the two groups. However, fibrin, inflammatory cell infiltration, multinucleated giant cell formation, endothelial disruption, and luminal distortion were greater in group I. On SEM, group I showed trans-mural penetration. In contrast, group II showed suture margin eversion and no transmural penetration. Stenosis was greater in group I than in group II on carotid angiogram. The operation time was shorter in group II than in group I, i.e. 5+/-1.4 min vs. 11+/-3.8 min, respectively. The current data showed similar or superior results with VCS clips in comparison to conventional suturing with polypropylene. PMID- 11410691 TI - The value of cystoscopy as an initial diagnostic modality for asymptomatic microscopic hematuria. AB - For the patients who visit outpatient clinics due to asymptomatic microscopic hematuria, cystoscopy has been looked upon as rather invasive compared to other diagnostic methods. We tried to elucidate the actual diagnostic value of cystoscopy in the initial evaluation of asymptomatic microscopic hematuria. We reviewed the results of cystoscopic examinations in 213 patients who visited our hospital due to asymptomatic microscopic hematuria. No definite lesion that could explain the microscopic hematuria was detected by means of IVP, urine cytology, and other nephrologic evaluations for all the patients. Among the abnormal cystoscopic findings in 55 patients, the lesions suspected to be directly related to microscopic hematuria were classified as 'significant lesions' (31 patients, 17.6%) which include entities such as bladder cancer (1.31%). 27 of 31 patients with significant lesions (85.2%) were over 50 yr old, and furthermore, 3 patients who were diagnosed as bladder tumor by cystoscopy were over 60 yr. Cystoscopy should be utilized as initial diagnostic modality in older patients with asymptomatic microscopic hematuria to rule out any possibility of bladder cancer occurrence. Further studies are needed to justify implementation of cystoscopy as an initial diagnostic modality in younger patients with asymptomatic microscopic hematuria. PMID- 11410692 TI - Effect of thyroid hormone on the alcohol dehydrogenase activities in rat tissues. AB - The effects of thyroid hormone on hepatic and gastric alcohol dehydrogenase (ADH) activities (nM of NADH/min/mg of cytosolic protein) have been investigated in male Sprague Dawley rats treated with thyroxine (1 mg/kg, po) for 14 days. Whereas hepatic ADH activity in thyroxine-treated rats decreased by 61.3% of control rats (26.4 vs 43.2, p<0.001), gastric ADH activity increased by 262.9% of control rats (4.9 vs 1.9, p<0.001). As for the activities of the lung and kidney, thyroxine treatment did not produce any statistically significant changes. These data suggest that thyrotoxicosis causes a decrease of hepatic alcohol metabolism, and that the increase of gastric ADH activity in thyrotoxic rats can partly restore the first-pass metabolism of ethanol. PMID- 11410693 TI - Immunohistochemical analysis of CD44s and CD44v6 in endometriosis and adenomyosis : comparison with normal, hyperplastic, and malignant endometrium. AB - The expression patterns of CD44s and CD44v6 were immunohistochemically compared with those of normal, hyperplastic and malignant endometrium. In normal endometria (n=37), endometrioses (n=46) and adenomyoses (n=20), the surface and glandular epithelial cells were negative for CD44s and CD44v6 in a proliferative pattern and positive in a secretory pattern, whereas the stroma was only positive for CD44s in both proliferative and secretory patterns. The endometrial hyperplasia (4 simple and 9 complex) had the identical patterns with normal proliferative phase of endometrium. Only one case showing complex hyperplasia with atypia was focally positive for CD44s and CD44v6 in glandular epithelia. CD44s and CD44v6 were positive in all endometrial adenocarcinomas (13), except one CD44s-negative case. In summary, the expressions of CD44s and CD44v6 in endometriosis and adenomyosis recapitulated those of normal cyclic endometrium. The expression patterns in endometrial hyperplasia were similar to those in normal proliferative endometrium, whereas the endometrial adenocarcinoma showed abnormal expressions for CD44s and CD44v6. Thus it was considered that the ectopic endometrium in endometriosis and adenomyosis was not aberrant as in endometrial carcinoma on the aspects of immunohistochemical expressions of CD44s and CD44v6. PMID- 11410694 TI - Metabolic characteristics and prevalence of osteoporosis among women in Tae-An area. AB - Understanding the metabolic changes in women is one of the important ways to prevent and treat osteoporosis. To reveal the metabolic characteristics of 289 healthy women aged between 35-65 yr in Tae-An, Korea we evaluated the association between bone mass assessed by broadband ultrasound attenuation (BUA) using quantitative ultrasound 2 (QUS2) and various parameters such as age, body mass index, serum levels of alkaline phosphatase, calcium, phosphorus, parathyroid hormone, 25(OH)D, and urinary ratios of calcium/creatinine and deoxypyridinoline (Dpyd)/creatinine. Among the subjects, 3.0% were osteoporotic, and 40.9% were osteopenic. When the subjects were classified according to their years since menopause (YSM) and age, the prevalence of osteoporosis increased along with an increase of YSM and age. Bone turnover markers such as serum alkaline phosphatase and fasting urinary Dpyd/creatinine were significantly higher in the group with low bone mass than in the normal group. In summary, this study shows, by use of biochemical markers of bone turnover and QUS2, the prevalence of osteoporosis in women aged between 35-65 in Tae-An was 3.0% and the risk of low bone mass increased with the bone turnover markers. PMID- 11410695 TI - 5-fluorouracil-induced leukoencephalopathy in patients with breast cancer. AB - The purpose of this study is to determine the characteristic clinical features, radiologic findings, and precipitating and prognostic factors in the patients with breast cancer and with 5-Fluorouracil (5-FU)-induced leukoencephalopathy. We reviewed the medical records of six breast cancer patients who developed leukoencephalopathy after chemotherapy which included 5-FU and also evaluated thorough neurological examinations including mini-mental status examination, cerebrospinal fluid studies, brain images and brain biopsies. Six patients exhibited slowly progressing neurologic symptoms characterized by the impairment of cognitive function, abulia, ataxic gait, and/or akinetic mutism. None of the patients had any specific causes or etiologic factors for leukoencephalopathy. Brain MRI in all patients showed diffuse periventricular white matter changes in the T2-weighted MR image. Brain biopsy in Patient 1 showed fragmented axonal fiber and minimally deprived myelination with many scattered macrophages. Five patients who treated with steroids at the onset of neurological symptoms showed clinical improvement, regardless of their age, sex, the pathology and stage of breast cancer, or the total dosage of chemotherapeutic agents. We conclude that leukoencephalopathy in these cases could be attributable to 5-FU neurotoxicity and suggest that the administration of steroids might be the treatment of choice. PMID- 11410696 TI - Effect of hypothermia on brain cell membrane function and energy metabolism after transient global hypoxia-ischemia in the newborn piglet. AB - This study was done to determine the effects of hypothermia on brain cell membrane function and energy metabolism after transient hypoxia-ischemia (HI) in the newborn piglet. Cerebral HI was induced by temporarily complete occlusion of bilateral common carotid arteries with surgical clips and simultaneous breathing with 8% oxygen for 30 min, followed by release of carotid occlusion and normoxic ventilation for 4 hr. Rectal temperature was maintained between 38.0 and 39.0 degrees C in normothermic groups, and between 34.0 and 35.0 degrees C in hypothermic groups for 4 hr after HI. During HI, heart rate, glucose and lactate level in the blood and cerebrospinal fluid increased, and base excess, pH and blood pressure decreased significantly in both normothermic and hypothermic groups. After HI, these abnormalities returned to normal in normothermic group, but lactic acidosis persisted in hypothermic group. Decreased cerebral Na+,K+- ATPase activity and increased lipid peroxidation products, indicative of HI- induced brain injury, were more profound in hypothermic group than in normothermic group. Brain ATP and phosphocreatine levels were not different between normothermic and hypothermic groups. In summary, hypothermia applied immediately after HI for 4 hr did not improve the recovery of brain cell membrane function and energy metabolism in the newborn piglet. PMID- 11410697 TI - Estimation of the incidence of head injury in Korea: an approximation based on national traffic accident statistics. AB - Head injury is a leading cause of death and disability in Korea. It usually results from an avoidable accident. Epidemiologic data on the head injury is important for the effective reduction of this controllable disaster. The aim of this study is to estimate the incidence of head injury in Korea. Data on the incidence of the traffic accidents were collected from Traffic Accident Statistics 1998 by the National Police Agency. Proportion of head injuries due to traffic accidents was obtained from various literatures. The incidence of head injury was approximated simply by a formula H=T/0.625 [the total number of patients after traffic accidents (T) would be 62.5% of the total number of patients with head injury (H)]. In 1998, the estimated number of head injury was 109,462. The annual incidence was 236/100,000 person, 334/100,000 for males and 136/100,000 for females. The causes were road traffic accident in 62.5%, and falls in 15.6%. The operation was performed in 20.2% with a mortality of 4.0% in average. Case fatality rate was 8.2%. Annual death rate was 19/100,000 population. A public campaign is required on the basis of trustworthy epidemiologic data to reduce the incidence of head injury for the public health. PMID- 11410698 TI - The effect of electrical stimulation on the trunk control in young children with spastic diplegic cerebral palsy. AB - This study was designed to determine the effectiveness of electrical stimulation over the trunk in improving sitting balance in young children with spastic diplegic cerebral palsy who displayed poor trunk control. The subjects ranged in age from 8 to 16 months and were randomly assigned to two groups. Both group had physical therapy for 6 weeks. Electrical stimulation (ES) group had additional electrical stimulation over the abdomen and posterior back muscles. Radiographic studies were carried out on the whole spine while they were sitting before and after treatment. Kyphotic angle, Cobb's angle and lumbo-sacral angle were measured. Additionally, sitting score-Gross Motor Function Measure (GMFM) was also evaluated. There was no difference of these values at initial evaluation between the two groups. Following 6 weeks of intensive therapy, the changes of kyphotic angle and sitting score-GMFM were significantly higher in ES group statistically when compared with those of the control group. The Cobb's angle following treatment was improved in ES group, but not statistically compared with that of control group. This study suggests that electrical stimulation over the trunk become a beneficial therapeutic technique in improving the sitting posture and trunk control in young children with spastic diplegic cerebral palsy. PMID- 11410699 TI - Systemic lupus erythematosus complicated with cerebral venous sinus thrombosis : a report of two cases. AB - A rare manifestation of systemic lupus erythematosus (SLE) is cerebral venous sinus thrombosis (CVST), in which early diagnosis and aggressive therapy are of prime importance for favorable outcome. The pathogenesis of CVST is largely unknown, but it is thought to be caused by cerebral vasculitis, antiphospholipid antibodies or other conditions associated with enhanced coagulability. We describe two cases of SLE with CVST which were not associated with antiphospholipid antibodies. Both cases were treated with immunosuppressants (intravenous methylprednisolone and cyclophosphamide pulse therapy) and anticoagulant drugs (heparin and subsequent maintenance therapy with warfarin). There was a marked improvement of neurologic symptoms with the disappearance of thrombus in a follow-up MRI. The possibility of CVST should be considered in any patients with SLE who show neuropsychiatric manifestations. PMID- 11410700 TI - A case of torsade de pointes associated with hypopituitarism due to hemorrhagic fever with renal syndrome. AB - We describe a 51-yr-old man presenting with syncope due to torsade de pointes. The torsade de pointes was refractory to conventional medical therapy, including infusion of isoproterenol, MgSO4, potassium, lidocaine, and amiodarone. His past history, physical findings, and hormone study confirmed that QT prolongation was caused by anterior hypopituitarism that developed as a sequela of hemorrhagic fever with renal syndrome. The long QT interval with deep inverted T wave was completely normalized 4 weeks after starting steroid and thyroid hormone replacement. Hormonal disorders should be considered as a cause of torsade de pointes, because this life-threatening arrhythmia can be treated by replacing the missing hormone. PMID- 11410701 TI - Emergence of vaccine-induced escape mutant of hepatitis B virus with multiple surface gene mutations in a Korean child. AB - The S protein of hepatitis B virus is the principal component of virus envelope and the primary target of anti-HBs response. Mutants or variants that escape neutralization by anti-HBs have been selected during immunoprophylaxis of HBV after birth and liver transplantation. We investigated a case of a Korean child who was vaccinated at birth against hepatitis B and also given hepatitis B immunoglobulin, but nevertheless later became infected with the virus. Hepatitis B virus-specific deoxyribonucleic acid covering the region of genome encoding the predominant "a" determinant of hepatitis surface antigen was amplified using polymerase chain reaction, and the nucleotide sequence was determined. We present for the first time in Korea the independent emergence of an escape mutant with substitution of arginine for glycine at amino acid 145 and proline for glutamate at amino acid 120 in "a" determinant after immunization. PMID- 11410702 TI - Restrictive cardiomyopathy in a patient with extrahepatic biliary atresia. AB - The most commonly associated anomalies in patients with extrahepatic biliary atresia are cardiovascular, digestive and splenic defects. Of the cardiovascular anomalies, there are very few reports of biliary atresia with cardiomyopathy. We report the first case of a child with extrahepatic biliary atresia and restrictive cardiomyopathy. The patient was a 13-month-old boy diagnosed with extrahepatic biliary atresia at the age of 2 months, when he underwent laparotomy for definite diagnosis.Hepatic portoenterostomy was performed after confirmative cholangiogram. Recently, he developed severe cough and dyspnea, and his respiratory symptoms worsened. Chest radiograph showed cardiomegaly. Two- dimensional echocardiography showed marked biatrial enlargement. On M- mode echocardiogram, a slight increase in left ventricular dimension was seen in early diastole with a relatively good left ventricular function. Mitral inflow Doppler tracing showed an increased E-velocity (1.1 m/sec) with decreased deceleration time (75 m/sec), and increased E/A ratio (0.33). He was diagnosed as having restrictive cardiomyopathy with characteristic echocardiographic features. PMID- 11410703 TI - Congenital monoblastic leukemia with 9;11 translocation in monozygotic twins : a case report. AB - We report an autopsy case of congenital monoblastic leukemia that developed in monozygotic twins. The twin presented with progressive hepatosplenomegaly at 4 weeks after birth. One twin died of massive bleeding and hypovolemic shock before the treatment started. At autopsy, the liver was diffusely enlarged and showed a diffuse whitish discoloration except for the subcapsular and perivenular areas. Microscopic examination disclosed infiltration of histiocyte-like atypical cells along the sinusoids and portal areas of the liver. Spleen, lymph nodes and choroid plexus were also infiltrated by the tumor cells. However, bone marrow involvement of the tumor was minimal although multifocal. On immunohistochemical staining, these atypical cells were reactive for CD68 (PGM-1) and lysozyme, suggesting that the tumor cells might have been derived from mono- histiocyte. Cytogenetic study revealed 9;11 translocation, which is frequently associated with acute monoblastic leukemia. To the best of our knowledge, this is the first report of congenital monoblastic leukemia of monozygotic twins in Korea. PMID- 11410704 TI - Retroperitoneal fibrosis with duodenal stenosis. AB - Retroperitoneal fibrosis is a rare disease characterized by the formation of dense plaque of fibrous tissue covering the retroperitoneal structures. This disease is commonly presented as ureteral obstruction, but the involvement of duodenum is rare. We report a case of retroperitoneal fibrosis which was complicated with duodenal stenosis and was successfully treated with corticosteroids. A 58-yr-old man, who had history of aorto-iliac bypass graft due to arteriosclerosis obliterans with infrarenal aortic occlusion was admitted to the hospital with abdominal pain and a mass. Abdominal CT scan revealed the periaortic soft tissue mass encircling grafted aorta and stenosis of duodenal third portion. Retroperitoneal fibrosis with duodenal stenosis was diagnosed and prednisolone therapy was initiated. Follow-up CT scan showed that the patient responded to prednisolone therapy with eased pain, shrinking periaortic mass, and reduced duodenal stenosis. PMID- 11410705 TI - Surgical treatment of digital ischemia occurred after radial artery catheterization. AB - Permanent ischemic injury of the hand after radial artery cannulation is rare, but several cases of thromboembolism after the cannulation leading to amputation of affected limb or digits have been reported. A 48-yr-old man undergoing spine surgery showed normal modified Allen's test and had no preoperative vascular disease. We inserted 20-G radial artery catheter for the continuous monitoring of the blood flow and serial blood sampling. There was no specific event during the operation and the catheter was removed immediately after the operation. The signs and symptoms of the circulatory impairment of the radial artery developed four days after the operation and aggravated thereafter. Through the angiographic study, we found the total occlusion of the radial artery and some of its branches. After an emergent surgical exploration of the radial artery for removal of the thrombus and vein graft for the defect of the artery on the 8th postoperative day, the ischemic signs and symptoms disappeared and the radial pulse was restored. PMID- 11410706 TI - A case of intramuscular sparganosis in the sartorius muscle. AB - Intramuscular sparganosis is not common, and its rarity makes it difficult to be distinguished from soft tissue tumors. A case of rare intramuscular sparganosis is reported. A 44-year-old man presented with a painful mass in the left thigh for 8 months, which was initially diagnosed a as soft tissue tumor. Ultrasonography and MRI revealed a multilobulated mass in the sartorius muscle. After the needle biopsy under the guidance of ultrasonography, sparganum was discovered under microscopic examination of the excised tissue. Surgical excision was performed, and a live larva of sparganum was removed. Sparganosis should be considered in the differential diagnosis of soft tissue tumors, especially among Koreans who have frequently ingested mountain water and consumed raw snakes or frogs. PMID- 11410707 TI - Functional analyses of a N-terminal splice variant of the alpha subunit of the epithelial sodium channel. AB - The amiloride-sensitive epithelial sodium channel (ENaC) is the limiting step for sodium absorption in epithelial cells of the distal nephron, distal colon, airways and excretory ducts of several glands. In vivo and in vitro studies showed that the alpha subunit of ENaC is necessary for the expression of functional channels. Using RT-PCR strategy, a novel N-terminal splice variant has been identified which deletes 49 amino acids in the N-terminal region of the mouse alphaENaC subunit. In oocytes expressing the alphaENaC splice variant, together with beta and gammaENaC subunits, amiloride-sensitive currents were less than 20% of values obtained with the wild type ENaC. The single channel conductance and the ionic selectivity were similar and there was only a minor decrease in the level of expression of the protein at the oocyte surface. These findings indicate that the deleted sequence in the N-terminal part of the mouse and rat alphaENaC subunit might play a role in the regulation of the activity of expressed ENaC channels. PMID- 11410708 TI - Regulation of Kv1.3 channels in activated human T lymphocytes by Ca(2+)-dependent pathways. AB - Activated T lymphoblasts respond more effectively to mitogenic stimuli than resting T cells, partly through differences in Ca(2+) signaling, which in turn depend on K(+) channel activity. Both Kv1.3 and Ca(2+)-activated K(+) (SK4) channels are up-regulated in T lymphoblasts. Since Ca(2+)- and calmodulin (CaM) dependent signal-ing are key pathways in T-cell activation, we investigated their involvement in regulating the Kv1.3 current. Kv1.3 in lymphoblasts was significantly inhibited by elevating internal Ca(2+) to the micromolar level. It was also reduced in a Ca(2+)-dependent manner by inhibiting CaM with W-7 or calmidazolium. Part of the CaM-dependence is likely through CaM kinase since the current was also inhibited by the antagonist, KN-62, but not by the inactive analogue, KN-04. Kinase inhibition, unlike CaM inhibition, was only effective at physiological temperatures, a difference that implies involvement of more than one mechanism. We demonstrated a biochemical association of Kv1.3 protein in lymphoblasts with the multifunctional type II CaM kinase, but not with calmodulin. Thus, Kv1.3 forms a multi-protein complex with CaM kinase II (which binds to Ca(2+)/CaM) and previously identified proteins (e.g., PSD-95, src tyrosine kinase) that position the channel to respond to signaling pathways that are crucial for T-cell activation and proliferation. PMID- 11410709 TI - PKA-induced stimulation of ROMK1 channel activity is governed by both tethering and non-tethering domains of an A kinase anchor protein. AB - We have used the patch-clamp technique to explore the role of A Kinase Anchor Proteins (AKAP) in mediating the effect of cAMP on ROMK1 channels expressed in the Xenopus oocytes. Addition of membrane permeant cAMP analogs increased channel activity only in oocytes injected with ROMK1 and AKAP79 cRNA but had no effect on channel activity in oocytes injected with ROMK1 alone. Using the two-electrode voltage clamp technique, we determined that application of H89, a potent inhibitor of protein kinase A (PKA), abolished the stimulatory effect of cAMP/forskolin. To investigate the role of AKAP specificity in conferring cAMP responses to ROMK1 channels, we examined channel activity in oocytes expressing ROMK1 and either AKAP18, AKAP-KL or AKAP75. Addition of cAMP failed to increase channel current in oocytes expressing ROMK1 and either AKAP18 or AKAP-KL. In contrast, cAMP increased ROMK1 channel activity by 33% in oocytes coexpressing AKAP75, the bovine homologue of AKAP79. The effect of cAMP on ROMK1 in oocytes coexpressing AKAP75 is inhibited by H89. Since all three AKAPs bind PKAII, the results suggest that a unique structural domain in AKAP75/79 collaborates with the PKAII binding site and enables a productive association of PKA with ROMK1 channels. Deletion of either the membrane targeting region of AKAP75 (AKAP45) or PKAII binding domain of AKAP75 (AKAP75DeltaC) abolished the effects of forskolin on ROMK1 channels. This suggests that the membrane targeting and the PKA binding domains of AKAP75 are essential for the effect of cAMP. However, the nature of the AKAP, that interacts with ROMK1 in the native tissue, remains to be determined because AKAP75/79 are not expressed in the kidney. We conclude that the regulation of ROMK1 channels by PKA requires the involvement of the cell membrane-directed AKAPs that are able to specifically link PKA to the target channel protein. PMID- 11410710 TI - K-Cl cotransport: immunohistochemical and ion flux studies in human embryonic kidney (HEK293) cells transfected with full-length and C-terminal-domain truncated KCC1 cDNAs. AB - Coupled K and Cl movements are mediated by several isoforms of the K-Cl cotransporter (COT) encoded by the KCC genes. The ubiquitous KCC1 isoform, important for cell volume and ion homeostasis, has 12 transmembrane domains (Tmds), and cytoplasmic N- and C-terminal domains (Ntd and Ctd). This study investigates the cellular localization of KCC1 by confocal microscopy, activation of K-Cl COT by various non-osmotic and osmotic interventions with net unidirectional K and Rb fluxes at 37( degrees )C, and the effect of Ctd deletion on K-Cl COT regulation. Human embryonic kidney (HEK293) cells were transfected with full-length (fl) rabbit (rb)KCC1 and - CtdKCC1 cDNAs obtained after truncation at nucleotide 2011. Normal cells exposed to polyclonal anti-Ctd antibodies against Ctd epitopes within a 77 amino acid sequence (a.a.943-1020) revealed granular membrane and cytoplasmic immunostaining, presumably endogenous KCC1. Additional diffuse membrane and cytoplasmic immunofluorescence in flKCC1 transfected cells was absent in -CtdKCC1-transfected cells. Monoclonal antibodies against a c-myc epitope at the protein Ntd showed both membrane and cytosolic fluorescence. Basal and N-ethylmaleimide (NEM)-stimulated Rb influxes through K Cl COT, calculated as Cl-dependent Rb fluxes, were 2-3-fold higher in flKCC1 transfected than in normal cells. NEM stimulation of K-Cl COT was highest in flKCC1-transfected cells, significantly lower in stably and abrogated in transiently -CtdKCC1-transfected cells. Furosemide, calyculin and genistein inhibited basal and NEM-stimulated K-Cl COT in normal and transfected cells. Staurosporine and hydroxylamine were ineffective stimulators. No effect of pH(0) changes (6.3-8.4) was observed in basal or NEM-stimulated K-Cl COT, in both normal and transfected cells. However, inhibition by NEM occurred at pH(0) 8.4. Furthermore, in a Cl-independent manner, NEM lowered cell K content by >30% and hypotonicity (210-70mOsM) stimulated furosemide-sensitive Rb influx and K loss. Thus, in cultured normal and KCC1-transfected cells, K-Cl COT shows significant differences from erythrocytes, and NEM and cell swelling open furosemide sensitive and Cl-independent K/Rb channels. Failure of K-Cl COT in cells transfected with Ctd-truncated KCC1 to respond to NEM suggests a role of the Ctd for signal transduction. PMID- 11410711 TI - Tyrosine kinases and phosphatases in the estivating spadefoot toad. AB - To endure seasonally arid conditions, spadefoot toads (Scaphiopus couchii) spend 9-10 months underground each year in a hypometabolic state, termed estivation. Protein tyrosine kinases (PTKs) and phosphatases (PTPs) were evaluated in organs of control and estivating toads to assess their possible role in signal transduction during estivation. Total PTK activity decreased by 27-52 % in liver, lung and skeletal muscle during estivation but rose by 66% in heart. Total PTP activity changed only in liver (55 % decrease) and heart (74 %increase). Analysis of the distribution of PTKs between cytoplasmic and membrane-associated forms showed that estivation-linked changes in both fractions occurred in heart (increase) and liver (decrease) whereas in lung and skeletal muscle only the soluble fraction was affected. PTPs were assessed using both a general substrate (ENDpYINASL) and a substrate (DADEpYLIPQQG) specific for PTPs containing the SH2 binding site; both revealed estivation-associated changes in activities and subcellular distribution of PTPs in all tissues. DEAE-Sephadex chromatography showed multiple forms of skeletal muscle PTKs and PTPs in both soluble and insoluble fractions. Each fraction showed three major peaks of PTK activity, two of which shifted in elution position during estivation. The data show that PTKs and PTPs are modified in an organ-specific fashion during estivation by three mechanisms: changes in total activity, changes in subcellular distribution and possible protein covalent modification. PMID- 11410712 TI - Genetic correlations between initial sensitivity to Ethanol and brain cAMP signaling in inbred and selectively bred mice. AB - BACKGROUND: Several lines of evidence have suggested a role for cAMP (adenosine 3',5'-cyclic monophosphate) signaling in the acute and chronic effects of ethanol. This study investigated whether there is a genetic correlation between cAMP synthesis in the brain and the acute effects of ethanol [alcohol sensitivity or acute functional tolerance (AFT)]. METHODS: By using nine inbred strains of mice, we measured initial sensitivity and AFT to ethanol with a test of balance on a dowel. Initial sensitivity was defined by the blood ethanol concentration (BEC0) at the loss of balance on a dowel after an ethanol injection [1.75 g/kg intraperitoneally (ip)]. When mice were able to regain balance on the dowel, BEC1 was determined, and a second ethanol injection was given (2 g/kg ip). Upon final regaining of balance, BEC2 was determined. AFT was defined by the difference between BEC1 and BEC2 (AFT = DeltaBEC = BEC2 - BEC1). Cyclic AMP synthesis was measured in whole-cell preparations in the cerebellum and other brain areas of mice of the nine inbred strains. RESULTS: Significant differences in BEC0 and AFT were seen among the mice of the nine inbred strains. Cerebellar basal and forskolin- and isoproterenol-stimulated cAMP production differed significantly between the strains, and BEC0 was found to correlate significantly with forskolin and isoproterenol-stimulated cAMP accumulation in the cerebellum (r = 0.70 and 0.94, respectively). When we measured cAMP production in mesencephalic and telencephalic tissue in three strains of mice that differed significantly in isoproterenol-stimulated cAMP accumulation in the cerebellum, significant differences between strains were found only in telencephalic tissue. The relative relationship between the rank order of the three strains for cAMP accumulation in the telencephalon and initial sensitivity to ethanol was identical to that seen with the cerebellum. However, AFT did not correlate with cAMP accumulation in the cerebellum or any other brain area tested. CONCLUSIONS: These results suggest that cAMP-generating systems of the cerebellum and possibly the brain areas contained in telencephalic tissues (e.g., basal ganglia) may have an important relationship to an animal's initial sensitivity to the incoordinating effects of ethanol. PMID- 11410713 TI - Genetic repeat polymorphism in the regulating region of CYP2E1: frequency and relationship with enzymatic activity in alcoholics. AB - BACKGROUND: Differences in the regulatory region of the CYP2E1 gene could be responsible for the interindividual variation in the cytochrome P-450 2E1 (CYP2E1) involved in ethanol oxidation. Recently, a polymorphic repeat sequence in the human gene was described between -2178 and -1945 base pairs. Its frequency seemed to vary among different ethnic populations, and it was suspected to be related to an increased inducibility to further ethanol intake. In the study reported here, the frequency of this polymorphism was investigated in a white French population. Its relationship with the previously described PstI/RsaI or DraI CYP2E1 polymorphisms, alcoholism, alcoholic liver disease, and inducibility of CYP2E1 by ethanol was examined. METHODS: The polymorphic region was characterized by polymerase chain reaction in 103 controls, 148 alcoholic subjects without liver diseases, and 98 others with liver cirrhosis. By using in vivo chlorzoxazone (CHZ) metabolism, CYP2E1 phenotype was assessed in 36 non ethanol-induced subjects (17 controls and 19 withdrawn alcoholics) and in 14 ethanol-induced subjects (10 controls after ingestion of 0.8 g/kg ethanol and four alcoholics with 100 g of daily intake). This phenotype was expressed as the 6-hydroxy CHZ/CHZ ratio. RESULTS: The rare allele frequency was found to be 1.58% in whites (n = 349). Neither significant association with alcoholism or alcoholic liver diseases, nor relationship with the PstI/RsaI polymorphism, was observed. But the DraI polymorphism was more frequent among the heterozygous subjects when compared with wild-type homozygous ones (p < 0.05). The CYP2E1 phenotype was similar in wild-type homozygotes and in heterozygotes at the constitutive level, as well as after induction with ethanol. CONCLUSIONS: Our data suggest that CYP2E1 repeat polymorphism does not seem to constitute a major factor for interindividual differences in CYP2E1 expression and susceptibility to alcohol related disorders in whites. PMID- 11410714 TI - No association between the dopamine d2 receptor taqi a1 allele and earlier age of onset of alcohol dependence according to different specified criteria. AB - BACKGROUND: The presence of the A1 allele of the dopamine D2 receptor TaqI restriction fragment length polymorphism has been reported to be associated with an earlier age of onset of alcohol dependence as a marker for severity. METHODS: We tested this hypothesis with special regard to the definition of the age of onset of alcoholism in 243 patients with alcohol dependence, according to DSM-IV criteria assessed by the standardized interview Munchner Composite International Diagnostic Interview (M-CIDI), consecutively admitted for detoxification. Additionally, the Addiction Severity Index (ASI) was performed. The TaqIA polymorphism was amplified by polymerase chain reaction (PCR), and the PCR product was digested by the restriction enzyme TaqI. Patients were subsequently divided into an A1 (presence of at least one A1 allele, n = 88) and an A2 group (absence of an A1 allele, n = 155). The following criteria for different definitions of age of onset were used: (1) age of onset of the first occurring symptom necessary for the diagnosis of alcohol dependence according to M-CIDI; (2) age of onset of the last symptom of alcohol dependence according to M-CIDI; (3) age of onset of more than 3 drinking days per week on a regular basis according to ASI; (4) age of onset of more than 3 drinking days-of more than five drinks per drinking day-or at least one binge drinking episode per week on a regular basis according to ASI. RESULTS: The frequency of the A1 allele in our patient sample was 0.208. No statistically significant association between the A1 allele and the age of onset of alcoholism was found. The mean age of onset according to criterion 1 was 30.4 +/- 10.8 years for the A1 group and 30.2 +/- 10.2 years for the A2 group (p = 0.89); for criterion 2, it was 33.3 +/- 10.0 years for the A1 group and 33.9 +/- 10.2 years for the A2 group (p = 0.77); for criterion 3, it was 18.0 +/- 7.5 years for the A1 group and 18.1 +/- 6.1 years for the A2 group (p = 0.92); and for criterion 4, it was 22.3 +/- 9.7 years for the A1 group and 21.8 +/- 8.5 years for the A2 group (p = 0.76). CONCLUSIONS: No association was found between the A1 polymorphism and age at onset of alcohol dependence according to different specified criteria. PMID- 11410715 TI - Comparison of global brain gene expression profiles between inbred long-sleep and inbred short-sleep mice by high-density gene array hybridization. AB - BACKGROUND: Inbred long-sleep (ILS) and short-sleep (ISS) mice show significant central nervous system-mediated differences in sleep time for sedative dose of ethanol and are frequently used as a rodent model for ethanol sensitivity. In this study, we have used complementary DNA (cDNA) array hybridization methodology to identify genes that are differentially expressed between the brains of ILS and ISS mice. METHODS: To carry out this analysis, we used both the gene discovery array (GDA) and the Mouse GEM 1 Microarray. GDA consists of 18,378 nonredundant mouse cDNA clones on a single nylon filter. Complex probes were prepared from total brain mRNA of ILS or ISS mice by using reverse transcription and 33P labeling. The labeled probes were hybridized in parallel to the gene array filters. Data from GDA experiments were analyzed with SQL-Plus and Oracle 8. The GEM microarray includes 8,730 sequence-verified clones on a glass chip. Two fluorescently labeled probes were used to hybridize a microarray simultaneously. Data from GEM experiments were analyzed by using the GEMTools software package (Incyte). Differentially expressed genes identified from each method were confirmed by relative quantitative reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: A total of 41 genes or expressed sequence tags (ESTs) display significant expression level differences between brains of ILS and ISS mice after GDA, GEM1 hybridization, and quantitative RT-PCR confirmation. Among them, 18 clones were expressed higher in ILS mice, and 23 clones were expressed higher in ISS mice. The individual gene or EST's function and mapping information have been analyzed. CONCLUSIONS: This study identified 41 genes that are differentially expressed between brains of ILS and ISS mice. Some of them may have biological relevance in mediation of phenotypic variation between ILS and ISS mice for ethanol sensitivity. This study also demonstrates that parallel gene expression comparison with high-density cDNA arrays is a rapid and efficient way to discover potential genes and pathways involved in alcoholism and alcohol related physiologic processes. PMID- 11410716 TI - Altered gabaa receptor subunit and splice variant expression in rats treated with chronic intermittent ethanol. AB - BACKGROUND: Intermittent chronic administration of ethanol to rats has been shown previously to produce a hyperexcitable, kindling-like state, accompanied by reduced inhibitory synaptic transmission in the hippocampus and changes in gamma aminobutyric acid type A (GABAA) receptors. Further information is needed on the detailed changes in GABAA receptors and their time course and persistence, as is comparison to changes after chronic, continuous ethanol. METHODS: GABAA receptors were analyzed in the rat brain after chronic intermittent ethanol (CIE) by using radioligand binding, photoaffinity labeling of polypeptides, and estimates of messenger RNA (mRNA) levels of receptor subunits by reverse transcriptase polymerase chain reaction (RT-PCR) and in situ hybridization. RESULTS: CIE rats were confirmed to have increased GABAA receptor binding of the benzodiazepine partial inverse agonist and ethanol antidote ligand Ro15-4513, due to increased expression of the alpha6 subunit polypeptide in the cerebellum, shown by photoaffinity labeling. Estimates of mRNA levels by use of RT-PCR did not reveal any significant increase in alpha6 or in several other receptor subunits in several brain regions, but a decrease in the ratio of the long and short splice variants (L/S) of the gamma2 subunit was detected in the hippocampus, especially the CA1 region. CONCLUSIONS: Changes in GABAA receptors were found in rats given CIE. Increased alpha6 subunit in the cerebellum was demonstrated by using both the binding to diazepam-insensitive sites for [3H]Ro15-4513 and increased levels of the 57-kDa alpha6 polypeptide after photoaffinity labeling with this ligand. This increase appeared after 30 doses of ethanol and decayed to normal 1 week after ethanol was discontinued. The transient change in cerebellar alpha6 subunit containing receptors, also reportedly seen after chronic continuous ethanol, is thus unlikely to account for the persistently hyperexcitable, kindled, seizure susceptible state seen in CIE. However, the significant decrease in gamma2 subunit L/S splice variant ratio in the hippocampus implies changes in GABAA receptor function, possibly involving protein phosphorylation by protein kinase C. Altered receptor trafficking and turnover associated with synaptic plasticity may contribute to the observed reduced inhibition in the hippocampus and other signs of alcohol dependence produced by CIE. PMID- 11410717 TI - 4-Methylpyrazole decreases salivary acetaldehyde levels in aldh2-deficient subjects but not in subjects with normal aldh2. AB - BACKGROUND: Carcinogenic acetaldehyde is produced from ethanol locally in the upper digestive tract via alcohol dehydrogenases (ADHs) of oral microbes, mucosal cells, and salivary glands. Acetaldehyde is further oxidized into less harmful acetate mainly by the aldehyde dehydrogenase-2 (ALDH2) enzyme. ALDH2-deficiency increases salivary acetaldehyde levels and the risk for upper digestive tract cancer in heavy alcohol drinkers. 4-methylpyrazole (4-MP) is an ADH-inhibitor which could reduce the local production of acetaldehyde from ethanol in the saliva. METHODS: Five ALDH2-deficient subjects and six subjects with normal ALDH2 ingested a moderate dose of alcohol (0.4 g/kg of body weight), whereafter their salivary acetaldehyde levels, heart rate, skin temperature, and blood pressure were followed for up to four hours. Blood acetaldehyde and ethanol levels were determined at 60 min. The experiment was repeated after a week. Two hours before the second study day, the volunteers received 4-MP, 10-15 mg/kg of body weight orally. RESULTS: Total ethanol elimination rate decreased with 4-MP by 38-46% in all subjects. 4-MP also reduced blood acetaldehyde levels and suppressed the cardiocirculatory responses of the ALDH2-deficient volunteers. In addition, salivary acetaldehyde production in ALDH2-deficient subjects was significantly reduced when correlated with salivary ethanol levels. On the contrary, 4-MP did not have any effect on salivary or blood acetaldehyde levels in subjects with normal ALDH2. CONCLUSIONS: A single dose of 4-MP before ethanol ingestion reduces ethanol elimination rate, the flushing reaction, and both blood and salivary acetaldehyde levels in ALDH2-deficient subjects but not in subjects with the normal ALDH2 genotype. These results suggest that the role of oral mucosal and glandular ADHs in salivary acetaldehyde production is minimal and support earlier findings indicating that salivary acetaldehyde production is mainly of microbial origin in subjects with normal ALDH2. PMID- 11410718 TI - Family interactions of alcoholics as related to alcoholism type and drinking condition. AB - BACKGROUND: Several studies have reported that the marital interactions of antisocial and aggressive, versus nonantisocial and nonaggressive, alcoholics exhibit higher rates of aversive-defensive communications and higher levels of negative reciprocity. To extend these findings, we examined the effect of alcoholism type (high- versus low-antisocial alcoholics: HAS, LAS) and drinking condition on family communication patterns. METHODS: Marital and parent-child dyads from 100 alcoholic families were videotaped while they discussed personally relevant issues during drinking and no-drinking sessions (no children were offered any alcohol). All interactions were coded with the Marital Interaction Coding System, and the data were assessed for differences in rate of positive, negative, and problem-solving behaviors, as well as sequential structures. RESULTS: HAS couples were more negative during the drink versus no-drink condition, whereas drinking did not affect negativity for LAS couples. In addition, the negative communications of HAS versus LAS alcoholics were more likely to increase spouse negativity during the drink versus no-drink condition. Group differences for parent-child interactions were few. CONCLUSIONS: The nature of family interactions was related to both alcoholism type and alcohol consumption, and the marital interactions of alcoholism types could be differentiated on the basis of the frequency and sequential structure of negative exchanges. It is most important to note that it is the interactions of the HAS alcoholic that undergo the most change as a function of drinking condition, with little support for the "adaptive consequences" hypothesis that alcohol consumption leads to more effective problem-solving for couples in which the alcoholic exhibits fewer antisocial and more internalizing characteristics. Study limitations and directions for future research are discussed. PMID- 11410719 TI - Effects of assessment frequency on subjective intoxication ratings after alcohol consumption. AB - BACKGROUND: It is common for subjective intoxication measures to be administered frequently throughout an experimental session. It is unclear, however, whether repeated assessments affect the experience of intoxication. This study examined the effect of assessing subjective intoxication levels during alcohol consumption on subsequent perceptions of intoxication after drinking. METHODS: Forty-two participants consumed a moderate dose of alcohol (men, 82 g/kg; women, 0.74 g/kg) during a 30-min period. Participants either reported or did not report subjective intoxication levels at 10-min intervals during the drinking period. After the drink, all participants rated their level of subjective intoxication on several occasions. RESULTS: Individuals who reported their intoxication during the drinking period reported higher levels after consumption than did those who did not rate their intoxication during drinking. CONCLUSIONS: These data suggest the potential for reactivity effects when conducting repeated assessments of perceived subjective intoxication. PMID- 11410720 TI - Combining ondansetron and naltrexone treats biological alcoholics: corroboration of self-reported drinking by serum carbohydrate deficient transferrin, a biomarker. AB - BACKGROUND: Recently, we showed by using self-report that combining ondansetron (4 microg/kg twice a day) and naltrexone (25 mg twice a day) was effective at reducing drinking and increasing abstinence among early-onset alcoholics (EOAs), who are characterized by a range of antisocial behaviors and high biological and familial disease predisposition. Here, we investigated whether the self-reported differences in drinking would be corroborated by measurements of serum carbohydrate-deficient transferrin (CDT) level, a sensitive, reliable, and well validated marker of transient alcohol consumption. METHOD: An 8-week double-blind clinical trial was performed in which 20 EOAs were randomized to receive ondansetron (4 microg/kg twice a day) and naltrexone (25 mg twice a day) or placebo as an adjunct to weekly standardized cognitive behavioral therapy. Serum CDT was assessed at weeks 0 (baseline), 4, and 8. RESULTS: Log serum CDT was significantly lower in the ondansetron and naltrexone group (group mean, 1.44 +/- 0.076) compared with the placebo group (group mean, 1.82 +/- 0.113), as evidenced by a main effect of group [F(1,15) = 7.2, p = 0.017; effect size = 0.32], visit [F(1,16) = 11.2, p = 0.004; effect size = 0.41], and an interaction between group and visit [F(1,16) = 27.54, p < 0.001; effect size = 0.63]. CONCLUSIONS: The combination of ondansetron plus naltrexone was superior to placebo at reducing serum CDT. This corroborated our self-reported drinking data and demonstrated that the medication combination is an effective treatment for EOAs. PMID- 11410721 TI - The identification of alcohol intoxication by police. AB - BACKGROUND: The identification of alcohol intoxication by police, bartenders, social hosts, and potential passengers is an important issue in the prevention of alcohol-related driving accidents. This study examines the ability of police officers to correctly identify and make ratings of the sobriety of target drinkers presented on video. METHODS: Raters were asked to determine (1) whether the target drinker had been drinking alcohol, (2) whether it was "okay" to serve the target another drink, and (3) whether the target drinker was "okay" to drive. A rater confidence score for each target evaluated, as well as demographic characteristics about the raters, was obtained. RESULTS: Drinkers were accurately targeted to low (0.08-0.09%), medium (0.11-0.13%), and high (0.15-0.16%) blood alcohol concentrations (BACs) by using a method previously described. At lower BACs, most police officers were unable to identify whether or not targets had been drinking. Raters were "pretty sure" that targets in the 0.15-0.16% range had been drinking and "not sure" whether or not they should be served another drink or drive a car. CONCLUSIONS: The ability of raters to reliably identify target drinkers who were too intoxicated to drive safely was not obtained until the BACs were relatively high. These results suggest that prevention measures must focus on improving behavioral observations made of potential drunk drivers. Implications for bartenders and social hosts are discussed. PMID- 11410722 TI - Ethanol-induced impairments in spatial working memory are not due to deficits in learning. AB - BACKGROUND: Acute ethanol administration impairs spatial reference memory and spatial working memory. However, the experimental designs previously used to test spatial working memory fail to make a distinction between the acquisition, or learning, of spatial information and the retention of this information. This study demonstrates that acute ethanol administration impairs spatial working memory, by using a novel experimental design that eliminates the confound between the learning of new spatial information and the testing of this information. METHODS: Long-Evans male rats received three forced trials to the same place for food reward on an elevated radial arm maze. Subjects were then given six free choice trials in the first acquisition session, followed by a 30-min consolidation period before an additional six free-choice trials were administered-the retention session. Animals were trained to a criterion of five of six correct in both the acquisition and retention over 2 consecutive days. Once criteria were obtained, subjects received either saline or one of three ethanol doses immediately after the acquisition session to investigate whether ethanol alters retention of the learned spatial information. RESULTS: Acute ethanol administration impaired spatial working memory. Rats tested under saline and low-dose ethanol (1.0 g/kg) made significantly more place choices than rats tested under moderate- or high-dose ethanol (1.5 and 2.0 g/kg, respectively). Ethanol produced a temporary impairment in that no significant differences were found when subjects were retrained and retested 24 hr after initial testing. CONCLUSIONS: These results demonstrate that acute ethanol administration impairs spatial working memory and that such a deficit is not contingent on a learning impairment. These results support earlier findings that acute ethanol administration impairs spatial working memory but provide a significant advance by validating a novel training procedure that allows for direct investigation of working memory. Ethanol's impairment of both spatial working memory and spatial reference memory strengthens the similarities between memory impairments due to ethanol administration and memory impairments due to hippocampal lesions. PMID- 11410723 TI - In utero ethanol exposure causes mitochondrial dysfunction, which can result in apoptotic cell death in fetal brain: a potential role for 4-hydroxynonenal. AB - BACKGROUND: In utero ethanol exposure causes abnormal fetal brain development that may partly be due to enhanced cell death. The mechanisms underlying this remain to be defined, but ethanol-induced oxidative stress may play a role. The following studies investigated the effects of short-term in utero ethanol exposure on fetal brain mitochondrial events that are known to elicit apoptotic cell death. Evidence is presented suggesting that 4-hydroxynonenal (HNE), a toxic product of lipid oxidation, is a causal factor in the observed mitochondrial damage. METHODS: Mitochondria were isolated from control and ethanol-exposed fetal brains (days 17 and 18 of gestation). Permeability transition was determined spectrophotometrically, and cytochrome c and apoptosis-inducing factor (AIF) release were assessed by Western blotting. Caspase-3 activity and DNA fragmentation were determined both as markers for mitochondrially mediated apoptosis and as consequences of cytochrome c and AIF release. RESULTS: Maternal ethanol intake caused an increase in mitochondrial permeability transition, and this was accompanied by cytochrome c and AIF release from fetal brain mitochondria that exceeded control values by 62 and 25%, respectively (p < 0.05). In utero ethanol exposure resulted in a 30% increase in caspase-3 activity and a 25% increase in DNA fragmentation (p < 0.05) in the fetal brain. HNE levels were increased by 23% (p < 0.05) in mitochondria by in vivo ethanol exposure. In vitro treatment of fetal brain mitochondria with HNE (25-100 microM) also caused increases in mitochondrial permeability transition, as well as dose-dependent releases of cytochrome c and AIF. CONCLUSIONS: These studies illustrate that in utero ethanol exposure can elicit a cascade of events in the fetal brain that are consistent with mitochondrially mediated apoptotic cell death. Additionally, the increase in mitochondrial content of HNE after ethanol intake and the ability of HNE added to fetal brain mitochondria to mimic these effects of in vivo ethanol exposure support a potential role for HNE in the proapoptotic responses to ethanol. PMID- 11410724 TI - Effect of alcohol consumption on host release of interleukin-17 during pulmonary infection with Klebsiella pneumoniae. AB - BACKGROUND: A link between alcohol abuse and bacterial pneumonia has been recognized for centuries, but mechanisms to explain this relationship are unclarified. Interleukin-17 (IL-17) is a lymphocyte-derived cytokine that is part of the inflammatory cytokine cascade. Previous studies from our laboratory indicated that IL-17 is released in lung tissue in a murine model of bacterial pneumonia caused by Klebsiella pneumoniae. The effects of alcohol consumption on pulmonary release of IL-17 are unknown. METHODS: Mice were maintained on 20% ethanol in drinking water or on a control diet without alcohol. After 2 weeks, alcohol and control mice were challenged with intratracheal K. pneumoniae. Mice were followed for survival after bacterial challenge, neutrophil recruitment was assayed as myeloperoxidase, and IL-17 was measured in lung lavage fluid by enzyme linked immunosorbent assay. In additional experiments, splenocytes from control mice were incubated with ethanol in vitro, and release of IL-17 was measured in culture supernatants. Finally, control and alcohol mice received intrapulmonary gene transfer of E-1-deleted adenovirus containing the murine IL-17 gene. These mice were then challenged with K. pneumoniae and followed for survival and neutrophil recruitment. RESULTS: In these studies, we demonstrate that a 2-week history of ethanol consumption in mice suppresses release of IL-17 into lung tissue, decreases neutrophil recruitment, and increases mortality from experimental K. pneumonia. In vitro experiments confirm a direct suppressive effect of ethanol on the release of IL-17 from splenocytes. In vivo administration of the IL-17 gene in an adenoviral vector to alcohol-consuming mice results in release of IL-17 into lavage fluid and normalizes neutrophil recruitment and mortality after bacterial challenge. CONCLUSIONS: The results of these experiments strongly implicate IL-17 as an important pathway for the immunosuppression associated with alcohol abuse and support gene therapeutic approaches to augment immune function in the alcoholic host or to treat infections associated with alcoholism. PMID- 11410725 TI - Cardioprotective effect of propranolol from alcohol-induced heart muscle damage as assessed by plasma cardiac troponin-t. AB - BACKGROUND: Heavy alcohol consumption from either long-term misuse or binge drinking is associated with poor cardiac contractility, mitochondrial dysfunction, and ventricular arrhythmias. The aim of this study was to measure circulating cardiac troponin-T as a marker for myocardial damage following acute and chronic alcohol administration. METHODS: In acute studies, male Wistar rats were treated with alcohol (75 mmol/kg body weight, intraperitoneal) and plasma was collected 2.5 hr after alcohol administration for analysis of rat cardiac troponin-T. In addition, rats were pretreated with cyanamide (an inhibitor of acetaldehyde dehydrogenase), various beta-blockers, xanthine oxidase inhibitors, or lisinopril before acute alcohol dosing. In chronic studies, rats were fed alcohol (as 35% of total dietary calories) for 6 weeks. RESULTS: The results of the time course study showed that acute alcohol administration significantly raised plasma cardiac troponin-T levels after 2.5 hr and 6 hr, but not after 24 hr. The effects of alcohol on cardiac troponin-T were potentiated with cyanamide pretreatment. Acute ethanol, alone or with cyanamide pretreatment, decreased systolic blood pressure and increased heart rates. Beta-blocker pretreatment with propranolol reduced the alcohol-induced increase in plasma troponin-T, whereas lisinopril potentiated this effect. The beta-blockers, atenolol and metoprolol, and the xanthine oxidase inhibitors, allopurinol and oxypurinol, were unable to reduce elevated troponin-T. However, pretreatment with the beta-blocker timolol moderated the acute alcohol-induced increase in troponin-T. In the chronic alcohol rat model, no differences were observed between alcohol and control pair fed rats, suggesting the inducement of tolerance. CONCLUSIONS: In conditions of acute exposure, ethanol-induced lesions are characterized by raised plasma cardiac troponin-T possibly due to beta1 and/or beta2 adrenergic activation. PMID- 11410726 TI - Effects of prenatal ethanol exposure on hypothalamic-pituitary-adrenal regulation after adrenalectomy and corticosterone replacement. AB - BACKGROUND: Previous studies have demonstrated that rats prenatally exposed to ethanol (E) exhibit hypothalamic-pituitary-adrenal (HPA) hyperresponsiveness, demonstrated by increased and/or prolonged elevations of adrenocorticotropin (ACTH) and/or corticosterone (CORT) after stress. This study investigated possible mechanisms of HPA hyperresponsiveness in E rats by manipulating CORT feedback regulation of HPA activity via adrenalectomy (ADX) with or without CORT replacement. METHODS: Male Sprague-Dawley rat offspring from prenatal E, pair-fed (PF) and ad libitum-fed control (C) groups were tested at 90 to 120 days of age. Rats were either sham-operated or underwent ADX, with or without CORT replacement. CORT (25 microg/ml) was replaced via the drinking water to achieve basal plasma CORT levels and maintain a phasic CORT signal. Seven days after surgery, animals were decapitated at the diurnal peak either under basal conditions or after a 15-min restraint stress, and trunk blood was collected. RESULTS: After ADX, loss of the CORT feedback signal resulted in increased plasma ACTH in all groups compared with those in sham animals. In addition, under basal conditions, ADX E rats had significantly greater plasma ACTH levels than both PF and C rats. However, no differences were seen in ADX rats after stress. CORT replacement after ADX was partially effective in normalizing ACTH levels under both basal and stress conditions, with no differences among E, PF, and C animals. CONCLUSIONS: These results suggest that E males may exhibit enhanced stimulatory inputs to the hypothalamus, increased pituitary sensitivity to secretagogues, or both, which may be revealed after ADX. In contrast, E animals seem similar to controls in their ability to use an exogenous CORT signal to regulate HPA activity. PMID- 11410727 TI - Mitochondrial dna damage and impaired mitochondrial function contribute to apoptosis of insulin-stimulated ethanol-exposed neuronal cells. AB - BACKGROUND: Ethanol inhibition of insulin signaling may contribute to impaired central nervous system development in fetal alcohol syndrome. An important consequence of ethanol inhibition of insulin signaling is increased apoptosis due to reduced levels of insulin-stimulated phosphoinositol-3-kinase activity. METHODS: We used viability assays, end-labeling, Western blot analysis, and MitoTracker (Molecular Probes, Eugene, OR) fluorescence labeling to determine whether ethanol-induced central nervous system neuronal cell death was mediated in part by increased mitochondrial (Mt) DNA damage and impaired Mt function. RESULTS: In ethanol-exposed, insulin-stimulated PNET2 central nervous system derived human neuronal cells, reduced viability was associated with increased Mt DNA damage, reduced Mt mass (manifested by reduced Mt protein expression and MitoTracker Green fluorescent labeling), and impaired Mt function (manifested by reduced levels of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide activity, cytochrome oxidase-Complex IV, Subunit II expression, and MitoTracker Red fluorescence). The adverse effects of ethanol on Mt function were reduced by pretreating the cells with broad-spectrum caspase inhibitors and nearly abolished by nerve growth factor stimulation, with or without concomitant treatment with global caspase inhibitors. CONCLUSIONS: These results suggest that ethanol induced death of insulin-stimulated immature neuronal cells is mediated in part by impaired Mt function associated with Mt DNA damage and reduced Mt mass, and therefore it is likely to contribute to neuronal loss associated with fetal alcohol syndrome. The findings also suggest that the adverse effects of ethanol on insulin-stimulated survival and metabolic function could be overcome by stimulating with growth factors that support Mt function through insulin independent pathways. PMID- 11410728 TI - Variations in corticosterone feedback do not reveal differences in hpa activity after prenatal ethanol exposure. AB - BACKGROUND: Prenatal ethanol exposure results in hypothalamic-pituitary-adrenal (HPA) hyperresponsiveness to stressors in adult animals. Possible mechanisms mediating this alteration in HPA responsiveness include stress-associated changes in corticosterone (CORT) feedback signals, alterations in CORT signals under basal conditions, and CORT-independent mechanisms. METHODS: We examined the effects of adrenalectomy (ADX) and CORT replacement with a constant, low-level CORT signal via CORT/cholesterol pellets on HPA responses to restraint stress. Adult Sprague-Dawley rats from prenatal ethanol (E), pair-fed (PF), and ad libitum-fed control (C) groups underwent sham ADX (sham), ADX without CORT replacement, or ADX with CORT replacement. Animals were tested during the trough of the circadian rhythm. RESULTS: In the sham condition, E females showed increased adrenocorticotropin hormone (ACTH) and CORT responses to restraint stress compared with C females. Basal and stress-induced ACTH levels were significantly increased in ADX compared with sham animals across all prenatal groups. Constant CORT replacement reduced basal ACTH levels compared with levels in the ADX group, although levels were still increased compared with those observed in the sham group. CORT replacement was minimally effective at reducing ACTH levels during stress. CONCLUSIONS: Although the effects of ADX may have masked possible influences of circadian drive or prenatal group, these findings suggest that in the absence of a CORT feedback signal or in the presence of a constant, low-level CORT feedback signal, E, PF, and C animals do not differ in their abilities to regulate ACTH secretion during the trough of the circadian rhythm. PMID- 11410729 TI - Effects of ethanol and naltrexone in a model of traumatic brain injury with hemorrhagic shock. AB - BACKGROUND: Ethanol predisposes to traumatic injury and causes respiratory depression and cardiovascular compromise in models of traumatic brain injury (TBI) and hemorrhagic shock (HS). Endogenous opioids may play a role in ethanol intoxication and TBI. We studied the effects of ethanol and the opiate antagonist agent naltrexone (NTX) in a TBI/HS model. METHODS: Fifty-six pigs (20 kg) were anesthetized with isoflurane, intubated, instrumented, and subjected to fluid percussion TBI with concurrent 30 ml/kg hemorrhage over 30 min. Seven groups were studied: Control, EtOH, NTX, INJ, INJ/EtOH, INJ/NTX, and INJ/EtOH/NTX. Ethanol (2 g/kg IV) was given preinjury, followed by infusion of 0.4 g/kg/hr. NTX 0.3 mg/kg intravenous was given 5 min postinjury. Parameters monitored for 120 min postinjury included minute ventilation (VE), blood pressure (MAP), cerebral perfusion pressure (CPP), cerebral venous lactate (Lac), arterial and cerebral venous blood gases, and brain tissue PtiO2. RESULTS: Ethanol levels at injury were 220 mg/dL. Ethanol-treated animals had depression of hypercapnic ventilatory response, which was reversed by administration of naltrexone. MAP and CPP were significantly lower in injured animals, but were not significantly improved by NTX. Cerebral venous pH was lower and lactate was higher in ethanol-treated animals. CONCLUSION: In this TBI/HS model, NTX reverses ethanol-induced depression of hypercapnic ventilatory response but does not improve MAP, CPP, or metabolic acidosis. This suggests that the respiratory effects of ethanol in TBI, but not the hemodynamic effects, may be mediated by opiate receptor activation. PMID- 11410730 TI - Chemical pathology in brain white matter of recently detoxified alcoholics: a 1H magnetic resonance spectroscopy investigation of alcohol-associated frontal lobe injury. AB - BACKGROUND: Investigations have suggested that frontal lobe abnormalities are a prominent feature of the alcoholic brain, indicated by impaired neuropsychological performance on tests of frontal lobe function and by reduced frontal lobe volume in neuroimaging and neuropathological examinations. White matter compartment volume loss may underlie observed brain shrinkage and cognitive deficits associated with the frontal lobes, although the nature of this change has not been well-characterized. METHOD: To investigate the susceptibility of frontal lobe white matter to alcohol-associated metabolic change and to understand the nature of alcohol-related white matter injury, 1H magnetic resonance spectroscopy (MRS) was used to measure concentrations of metabolites in frontal white matter (FWM) and parietal white matter (PWM) of recently detoxified alcoholics (RDA) and nonalcoholic controls (CON). Concentrations of N acetylaspartate (NAA), choline-containing compounds (Cho), myo-inositol (Ins), and creatine plus phosphocreatine (Cr) were measured in 37 RDA (mean age, 40.4 years; mean length of abstinence, 27.9 days) and 15 CON (mean age, 38.0 years). RESULTS: Analysis of variance (ANOVA) revealed a group by region of interest interaction for concentrations of NAA. Simple effects analysis revealed a significant 14.7% reduction in FWM NAA, while NAA levels in PWM were similar in RDA and CON. In addition, RDA had an 11.8% increase (averaged across both regions of interest) in brain white matter Ins relative to CON. Reductions in FWM NAA were associated with a longer drinking history in the RDA group, but this result was not found when both age and drinking history were used to predict the level of FWM NAA. CONCLUSIONS: Alcohol-associated reductions in FWM NAA may be the result of neuronal loss or dysfunction in the metabolism of NAA. While alcohol induced oxidative stress may cause global brain impairments in the metabolism and subsequent reduction of NAA, the frontal lobes are particularly rich in excitatory amino acid pathways, and axonal damage or destruction secondary to glutamate-mediated excitotoxicity during alcohol withdrawal may cause frontal lobe-specific reductions in NAA. Elevations in brain white matter Ins may reflect astrocyte proliferation as well as an osmotic response to cell shrinkage. PMID- 11410731 TI - Inhibition of caspases in vivo protects the rat liver against alcohol-induced sensitization to bacterial lipopolysaccharide. AB - BACKGROUND: The mechanisms of liver sensitization by alcohol to Gram-negative bacterial lipopolysaccharide (LPS) remain elusive. The purpose of this study was two-fold: (1) to test the hypothesis that alcohol-enhanced liver apoptosis may be a sensitizing mechanism for LPS and (2) to further characterize the liver apoptotic response to alcohol. METHODS: Rats were fed a high-fat, alcohol containing liquid diet for 14 weeks, treated with LPS (1.0 mg/kg of body weight, intravenously) or saline, followed by injection of a pan-caspase inhibitor IDN1965; N-[(1,3-dimethylindole-2-carbonyl)-valinyl]-3-amino-4-oxo-5 fluoropentanoic acid; 10 mg/kg of body weight, intraperitoneally or vehicle, and killed. The following parameters were assessed: plasma aspartate: 2-oxoglutarate aminotransferase activity (AST); liver histology and terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) response; caspase-3, -8, and -9 activity; and mRNA and protein expression for two apoptosis-signaling molecules: Fas receptor and Fas ligand; and three apoptosis adaptors: Bax, Bcl-XL, and Bcl-2. RESULTS: Alcohol-feeding-induced liver steatosis, slightly increased caspases' activity, the number of TUNEL-positive nuclei, and facilitated the LPS necrotic effect without affecting mRNA expression of apoptosis signals and adaptors. LPS induced a significant increase in AST and the number of TUNEL-positive nuclei, both effects being more pronounced in alcohol-treated rats. LPS produced hepatic necrosis only in alcohol-treated rats. LPS effects were associated with up-regulation of mRNA expression for both apoptosis adaptors and signaling molecules. IDN1965 administration 3 hr after LPS injection strongly inhibited caspases' activity, particularly that of caspase-3. IDN1965 also abolished the increase in TUNEL-positive nuclei, reversed the effect of LPS on plasma AST in alcohol-treated rats, and prevented LPS-induced necrosis. CONCLUSIONS: (1) Alcohol-enhanced liver apoptosis may not involve regulatory steps at the transcriptional level. LPS-induced liver apoptosis seems to involve transcriptional regulation of several apoptosis adaptors. Therefore, alcohol and LPS may enhance liver apoptosis through different mechanisms. (2) Alcohol enhanced liver apoptosis precedes and may facilitate the hepatic effects of LPS. LPS superimposed on alcohol further elevates the rate of apoptosis in the liver. This may exceed the phagocytosing capacity of the liver so that all the apoptotic cells are not phagocytosed, but rather die of necrosis. PMID- 11410732 TI - Study of cytochrome P4502E1 mRNA level of mononuclear cells in patients with alcoholic liver disease. AB - BACKGROUND: Cytochrome P-4502E1 (CYP2E1) is an important enzyme because of its unique ability to convert many substrates to cytotoxins. The increased production of reactive intermediates by elevated enzyme concentrations leads to various pathological conditions. Therefore, it is important to detect induced CYP2E1 levels in alcoholic individuals to avoid xenobiotic-promoted liver injury. In the present investigation, we detected CYP2E1 mRNA levels of mononuclear cells obtained from 10 ml of blood by using competitive polymerase chain reaction (PCR) method. METHODS: Mononuclear cells were obtained from healthy individuals who did and did not drink habitually and patients with alcoholic liver disease (ALD). Complementary DNA synthesis was performed with RNA obtained from mononuclear cells by reverse transcription-PCR. Competitive PCR of CYP2E1 was performed with the sense (5'-CTGCAACGTCATA-GCCGACA-3') and antisense (5'-TCCATTTCCACGAGCAGGCA 3') primer and competitor DNA. Competitive PCR of beta-actin also was performed. Electrophoresis was scanned, and each band was digitized. The concentration of CYP2E1 and beta-actin mRNA was calculated from the ratio of competitor DNA. RESULTS: In healthy individuals who did and did not drink habitually, CYP2E1 mRNA levels were 103.3 copies/microl RNA and 101.7 copies/microl RNA, respectively. In actively drinking patients with ALD, CYP2E1 mRNA levels were 103.5 copies/microl RNA, but those levels decreased to 101.7 copies/microl RNA after 4 days of abstinence. No significant difference was observed in CYP2E1 mRNA levels between alcoholic fibrosis and cirrhosis. As control, we measured beta-actin mRNA levels in mononuclear cells in all samples. The mean value of beta-actin mRNA was 104.3 copies/microl RNA in all cases, which included patients with ALD. CONCLUSIONS: The results demonstrated that it is possible to measure the CYP2E1 mRNA levels of mononuclear cells in a 10 ml blood sample. The CYP2E1 mRNA level in mononuclear cells increases during drinking and decreases in abstinence for a short period of 3 to 4 days. It is concluded that CYP2E1 mRNA level may be used as an effective marker for alcoholic intake. PMID- 11410733 TI - Polymorphism of tumor necrosis factor-beta and alcohol dehydrogenase genes and alcoholic brain atrophy in Japanese patients. AB - BACKGROUND: Alcohol abuse can induce brain atrophy, but it only occurs in some alcoholics. Many inflammatory cytokines such as tumor necrosis factor (TNF) are produced rapidly in the brain by experimental or clinical injury. METHOD: To investigate whether genetic polymorphism of TNF was related to alcoholic brain atrophy, we determined restriction fragment-length polymorphisms of the TNF-beta genes in 72 male alcoholics. Computed tomography was used to determine the severity of brain atrophy. RESULTS: Digestion with NcoI and MspI after polymerase chain reaction amplification showed that the TNFB1 allele frequency was significantly higher in patients with brain atrophy than in those without brain atrophy (chi2 = 10.20, p = 0.0034). A multivariate analysis that included age, total alcohol intake, ADH2 genotype, and TNF-beta genotype showed that the ADH21/21 genotype and TNFB1/B1 genotype are independently associated with alcoholic brain atrophy. These findings suggest that the TNFB1 allele may be associated with alcoholic brain atrophy. PMID- 11410734 TI - Polymorphism of the 5'-flanking region of the CYP2E1 gene: an association study with alcoholism. AB - BACKGROUND: Recently, a restriction fragment length polymorphism in the regulatory region of the CYP2E1 gene was identified. It has been suggested that the polymorphism is associated with the elevated activity of the cytochrome P-450 2E1 (CYP2E1) enzyme in obese or alcoholic subjects. However, significance of the polymorphism in connection with alcoholism has not been studied. In the present study, we have characterized these repeated sequences in the 5'-untranslated region of the CYP2E1 gene in Japanese subjects and North American white subjects and investigated whether these polymorphisms are associated with drinking habits and alcoholism. METHODS: DNAs were isolated from blood samples of 192 Japanese nonalcoholics and 202 alcoholics as well as 125 North American white nonalcoholics. DNA samples were amplified by polymerase chain reaction and subjected to a fluorescent-based single-strand conformational change polymorphism analysis, DNA fragment analysis, and polymerase chain reaction-direct sequencing. RESULTS: Four alleles (A1-A4) were found, which were differentiated by the six subunits (L1, L2, L3, L4, S1, S2) based on the size difference and nucleotide replacement. A2 and A4 alleles were observed in the two ethnic groups (A2: Japanese subjects, 0.752; white subjects, 0.976; A4: Japanese subjects, 0.227; white subjects, 0.016). However, A1 allele was found only in North American white subjects (0.008), and A3 allele was detected only in Japanese subjects (0.021). Allele frequencies were significantly different between the two ethnic groups (p < 0.0001). Distribution of genotypes between Japanese nonalcoholics and alcoholics were not significantly different. Also, no significant difference for the allele frequencies was observed between Japanese moderate drinkers and heavy drinkers. CONCLUSIONS: Our data suggested no association among the polymorphic repeats, drinking behavior, and alcoholism. Allele frequencies were significantly different between Japanese subjects and North American white subjects. PMID- 11410735 TI - Association of polymorphism in the alcohol dehydrogenase 2 gene with alcohol induced testicular atrophy. AB - BACKGROUND: Alcohol abuse can induce testicular atrophy, but it only occurs in some alcoholics. Alcohol dehydrogenase (ADH) is located principally on the Leydig cells. METHODS: To investigate whether genetic polymorphism of alcohol dehydrogenase (ADH) 2 and aldehyde dehydrogenase (ALDH) 2 was related to alcoholic testicular atrophy, we determined restriction fragment-length polymorphisms of the ADH2 and ALDH2 genes in 43 Japanese male alcoholics and 50 healthy subjects. An orchidometer was used to determine the testicular size. RESULTS: Less than 16 ml in testicular size was defined as testicular atrophy. Testicular atrophy was found in 24 (55.8%) cases out of 43 alcoholics. Digestion with MaeIII and MboII after polymerase chain reaction amplification showed that the ADH21 allele frequency was significantly higher in patients with testicular atrophy than in those without testicular atrophy (chi2 = 4.665, p = 0.031), whereas no significant association was observed between testicular atrophy and the ALDH2 gene. CONCLUSIONS: The ADH21 allele may be associated with alcoholic testicular atrophy. PMID- 11410736 TI - Effect of ALDH2 and CYP2E1 gene polymorphisms on drinking behavior and alcoholic liver disease in Japanese male workers. AB - AIMS: We examined the relationships of ALDH2 and CYP2E1 genotypes on drinking behavior and the incidence of alcoholic liver disease in Japanese male workers. METHODS: Two hundred and eighty-seven Japanese men were selected from one metal company to adjust for similar economic and social backgrounds. Drinking behavior was assessed from a self-assessment questionnaire. Genotypes of ALDH2 and CYP2E1 were analyzed with the polymerase chain reaction-single strand conformation polymorphism and with the polymerase chain reaction-restriction fragment length polymorphism, respectively. RESULTS: The frequency of the ALDH2 genotype was 55% for typical homozygotes, 42% for heterozygotes, and 4% for atypical homozygotes. The frequency of the CYP2E1 genotype was 62% for c1 homozygotes, 35% for heterozygotes, and 3% for c2 homozygotes. The ALDH2 genotype closely influenced drinking habits, but not the CYP2E1 genotype. Among habitual drinkers, ALDH2 typical homozygotes consumed significantly larger amounts of ethanol than ALDH2 heterozygotes, whereas CYP2E1 genotypes did not influence daily alcohol consumption. Sixteen men (5.6%) were diagnosed with alcoholic liver disease. In terms of ALDH2 genotypes, 12 cases (7.6%) were typical homozygotes and 4 (3.4%) were heterozygotes, whereas the incidence of alcoholic liver disease was not different between c1/c1 homozygotes and c1/c2 heterozygotes. When the interactive contribution of the ALDH2 and CYP2E1 genotypes on drinking behavior and the incidence of alcoholic liver disease were examined, there were no significant differences in the CYP2E1 genotype among the subjects with the same ALDH2 genotype. CONCLUSION: The ALDH2 genotype is strongly associated with individual alcohol drinking behavior and the development of alcoholic liver disease in Japanese male workers, but the CYP2E1 genotype is not. PMID- 11410737 TI - Nicotinamide adenine dinucleotide-dependent retinoic acid formation from retinol in the human gastric mucosa: inhibition by ethanol, acetaldehyde, and H2 blockers. AB - All-trans retinoic acid formation from all-trans retinol (vitamin A) in the human gastric mucosa was studied. When all-trans retinol and the human gastric mucosa were incubated together, all-trans retinoic acid was formed in the presence of nicotinamide adenine dinucleotide (NAD). When the NAD was not added, hardly any formation was observed. The formation of all-trans retinoic acid tended to be attenuated by 10 mM ethanol. Moreover, it was significantly attenuated in a concentration-dependent manner by ethanol at concentrations of 100 mM and above. Acetaldehyde at concentrations of 50 microM and above also significantly attenuated its formation in a concentration-dependent manner. Some H2 blockers, which include ranitidine hydrochloride and cimetidine, significantly attenuated the formation of all-trans retinoic acid, whereas famotidine failed to suppress it. There is an NAD-dependent pathway by which all-trans retinoic acid is produced from all-trans retinol in the human gastric mucosa. Inhibitors of alcohol dehydrogenase, which include ethanol and some H2 blockers, and of aldehyde dehydrogenase, which include acetaldehyde, inhibit its production. PMID- 11410738 TI - Alcohol dehydrogenase activities in the human gastric mucosa: effects of Helicobacter pylori infection, sex, age, and the part of the stomach. AB - BACKGROUND: Human gastric mucosa contains three alcohol dehydrogenase (ADH) isozymes (classes I, III, and IV). Various factors such as Helicobacter pylori infection, sex, age, and the part of the stomach involved have been suggested to affect alcohol dehydrogenase activities, although these views are controversial. In this study, these unsettled issues were reexamined. METHODS: Activities of class I and IV ADHs were evaluated in the cytosolic fraction of human gastric mucosa samples by reduction of their preferred substrates, namely acetaldehyde and m-nitrobenzaldehyde, and activities of class III were evaluated by oxidation of its preferred substrate, formaldehyde. Then, effects of Helicobacter pylori infection, sex, age, and the part of the stomach involved were examined. RESULTS: Class I, III, and IV ADH activities were 17.5 +/- 8.4, 4.2 +/- 2.5, and 8.9 +/- 3.9 nmol of nicotinamide adenine dinucleotide oxidation per minute per milligram of protein, respectively, for the entire population. Helicobacter pylori infection significantly reduced class I and IV ADH activities but did not affect activity of class III. In the samples without Helicobacter pylori infection and severe gastritis, sex did not affect class I, III, or IV ADH activities. In the same series, class IV ADH activity significantly decreased with age (p = 0.006), whereas no correlation was found between age and ADH activity of class I and III ADHs. The level of class IV ADH activity was significantly higher in the upper body than in the lower regions, whereas no such heterogeneity was observed in class I and III ADH. CONCLUSIONS: Various factors affect human gastric ADH activities, such that careful interpretation of their significance is necessary. PMID- 11410739 TI - Effects of chronic alcohol consumption on hepatic poly-ADP-ribosylation in the rat. AB - BACKGROUND: Poly-adenosine diphosphate (ADP)-ribosylation is involved in a variety of biological processes, which include DNA repair, malignant transformation, and apoptosis. It is of interest how this reaction is altered after long-term alcohol intake. Therefore, we determined long-term alcohol effects on hepatic poly-ADP-ribosylation in the rat. METHODS: Male Sprague Dawley(R) rats (four pairs) were pair-fed a nutritionally adequate liquid diet that contained ethanol as 36% of total energy and an isocaloric control diets for 4 weeks. Liver tissue homogenates and nuclear fractions were subjected to ADP ribosylation with [32P]nicotinamide adenine dinucleotide. The ADP-ribosylated proteins were separated by SDS-PAGE, followed by autoradiography. Expression of poly-ADP-ribose polymerase (PARP) also was evaluated by Western blotting. RESULTS: Incubation of rat liver homogenates in ADP-ribosylation reaction mixture resulted in a radiolabeling of a 116 kDa protein, most likely auto-ribosylation of PARP. This poly-ADP-ribosylation was increased significantly (p < 0.025) after long-term alcohol intake. This alcohol effect was reproducible in nuclear fractions as well. Expression levels of PARP, however, were comparable between alcohol-fed rats and their pair-fed controls. CONCLUSION: Poly-ADP-ribosylation, an important posttranslational modification of nuclear proteins, was increased significantly after chronic alcohol consumption in the rat. PMID- 11410740 TI - Effects of ethanol on L-arginine transport in rat Ito cells in relation to nitric oxide production. AB - BACKGROUND: Nitric oxide (NO) is a potent mediator of hepatic sinusoidal hemodynamics that is synthesized in the hepatic stellate cells (Ito cells, fat storing cells) and affects these cells. NO production may depend on the induction of inducible nitric oxide synthase and on transport of extracellular L-arginine. The precise mechanism that controls NO production in stellate cells was characterized recently. METHODS: Kinetic analysis of L-arginine transport and reverse transcription-polymerase chain reaction for cationic amino acid transporter (CAT) were carried out by using stellate cells prepared from the male Wistar rat. The effect of ethanol on L-arginine transport and NO production of stellate cells was assessed in the presence of tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma. RESULTS: The L-arginine transport system functioning in the hepatic stellate cells was system y+, possibly mediated by CAT-1 and CAT 2B (Km approximately 50 microM). IFN-gamma in combination with TNF-alpha induced NO production with an enhancement in CAT-2B mRNA expression and L-arginine transport, whereas L-arginine transport and NO production were suppressed by coincubated ethanol. CONCLUSIONS: In hepatic stellate cells, ethanol has suppressive effects on NO production and extracellular L-arginine transport in the presence of TNF-alpha and IFN-gamma. The estimated Km of L-arginine transporter in hepatic stellate cells is very similar to the physiological L arginine concentration in portal vein. Our findings may support the merit of further studies on the modulation of NO production via access to portal blood L arginine concentration to control disturbed hepatic sinusoidal blood flow in patients with alcoholic liver disease. PMID- 11410741 TI - A variant of des-gamma-carboxy prothrombin was increased in alcoholic liver disease without hepatocellular carcinoma. AB - Serum variants of des-gamma-carboxy prothrombin (DCP) recognized by two different monoclonal antibodies, 19B7 and MU-3, were measured in patients with alcoholic liver disease (ALD), and the values were compared with those of viral liver disease (VLD) and hepatocellular carcinoma (HCC). In the assay that used 19B7 antibody, DCP levels in ALD and HCC were significantly higher than that of VLD, although there was no significant difference in the values between ALD and HCC. In the assay that used MU-3 antibody, DCP level of HCC was significantly higher than those of ALD and VLD, although there was no significant difference in values between ALD and VLD. The ratio of 19B7/MU-3 assay values was significantly higher for ALD than the ratios for VLD and HCC. It is suggested that ALD has a different DCP variant pattern compared with VLD and HCC, which suggests that ALD has a different mechanism of DCP production. PMID- 11410742 TI - Kupffer cell sensitization by alcohol involves increased permeability to gut derived endotoxin. AB - BACKGROUND: Studies with gut sterilization and Kupffer cell inactivation support the hypothesis that endotoxin and Kupffer cells are involved in mechanisms of alcohol-induced liver injury. Recently, we found that Kupffer cells isolated from rats treated only once with ethanol were sensitized to endotoxin 24 hr later. Moreover, we established a new, simple animal model of ethanol hepatotoxicity based on Kupffer cell sensitization. The purpose of this study was to determine the mechanisms by which alcohol sensitizes Kupffer cells to lipopolysaccharide (LPS). METHODS: Female Wistar rats were given ethanol (5 g/kg body weight) once every 24 hr intragastrically, and ethanol concentration, ethanol elimination, and portal vein endotoxin were measured. Gut permeability was measured in isolated segments of ileum by translocation of horseradish peroxidase. Kupffer cells were isolated 24 hr after ethanol administration in vivo and were cultured in RPMI 1640 with 10% fetal bovine serum. After the addition of LPS, intracellular Ca2+ was measured by using a microspectrofluorometer with the fluorescent indicator fura-2, and tumor necrosis factor (TNF)-alpha was measured by enzyme-linked immunosorbent assay. CD14 was evaluated by Western analysis. RESULTS: Ethanol levels exhibited a cyclic pattern in ethanol-treated rats. Similar results were obtained in groups given ethanol and antibiotics for 4 weeks. Rates of alcohol elimination were around 3.5 mmol/kg/hr in control rats. After 4 weeks of ethanol treatment with or without antibiotics, elimination rates were not changed. Translocation of horseradish peroxidase was increased about 3-fold in gut segments by treatment with ethanol. This increase was not altered by treatment with antibiotics. Moreover, portal vein endotoxin levels were increased from nearly undetectable levels to 80 pg/ml in plasma of rats treated with ethanol. As expected, this increase was prevented (<20 pg/ml) by antibiotics. In isolated Kupffer cells from rats treated with ethanol for 4 weeks, CD14, LPS-induced intracellular Ca2+, and TNF-alpha all were increased. These phenomena were blocked by antibiotics. CONCLUSIONS: Kupffer cells isolated from rats treated with ethanol for 4 weeks exhibit sensitization to LPS. It is likely that increased permeability of the gut is a prominent event that leads to alcoholic liver injury. PMID- 11410743 TI - Alteration of annexin IV expression in alcoholics. AB - Western blot analysis was performed by using a specific antibody to measure annexin IV in human postmortem brain samples from alcoholic subjects. The analysis showed a significantly augmented expression in the hippocampus compared with controls, whereas the expression in the frontal cortex was equivalent in both groups. Annexin IV expression in the occipital cortex tended to increase in alcoholics. It was shown further that autoantibodies to annexin IV were increased significantly in alcoholic patients compared with controls. Thus, annexin IV may become a novel biological marker for alcoholics. PMID- 11410744 TI - Polymorphism of the neuropeptide Y gene: an association study with alcohol withdrawal. AB - BACKGROUND: Neuropeptide Y (NPY) exists in both the central and peripheral nervous system and is thought to modulate many functions such as feeding behavior, anxiety-associated behavior, circadian rhythm, seizure modulation, and hormone secretion. Recent studies have revealed that NPY influences alcohol consumption in mice and that alcohol-preferring rats showed lower concentrations of NPY-like immunoreactivity compared with alcohol-nonpreferring rats in several brain regions. METHODS: In the present study, we analyzed the whole coding region and 5'-untranslating region of the NPY gene for 163 Japanese male alcoholics with different withdrawal symptoms (93 with delirium tremens, 71 with seizures, 49 with hallucinations) and 98 Japanese male controls. ALDH2 genotypes of all subjects tested were ALDH2*1 homozygote. RESULTS: Three polymorphic nucleotide substitutions, namely -121C/A (promoter), 1258G/A (exon 2), and 5671C/T (exon 3), were detected in both groups. Substituting C to A in the -121 locus produced a putative binding site of GATA-1 and GATA-2. Also, -90G/A (promoter) as a rare variant and 5642-5651 single nucleotide repeats T10/T11 (intron 2) were found. Polymorphism (C/T) at the 1128 locus has been reported to be associated with a higher serum cholesterol level in obese white subjects, but such a polymorphism was not found in our samples. The genotypical distributions for these polymorphic loci (-121C/A, 1258G/A, 5642-5651 T10/T11, and 5671C/T) were not significantly different between the alcoholics and controls. However, frequency of the T allele and frequency of the genotype that possessed T alleles (CT, TT) at the 5671 locus were significantly higher in patients with seizure than in those without seizure (p < 0.05, p < 0.02). CONCLUSION: Our data suggested that a C to T substitution at the 5671 locus of the NPY gene may be associated with seizure during alcohol withdrawal. PMID- 11410745 TI - Electroacupuncture stimulation suppresses the increase in alcohol-drinking behavior in restricted rats. AB - BACKGROUND: Although a number of studies on traditional eastern or Chinese medicine, such as acupuncture, moxibustion, and herbal drugs, have been reported, few reports describe electroacupuncture (EAC) effects on drug- and alcohol seeking behaviors in animal models. The purpose of the present study was to investigate the effect of EAC on changes in alcohol-drinking behavior in rats challenged with restriction and immobilization stress. MATERIAL AND METHODS: Male Sprague Dawley rats (260-280 g) were tightly hung and immobilized in restriction models for 10 min. These immobilization stresses were performed twice a week for 1 week and for 3 consecutive weeks for the short- and long-restricted stress groups, respectively. EAC was applied for 10 min to the hindlimb point, Tsu-San Li (ST 36), and the lumbar point, Shen-Shu (BL 23). These points are used to treat mental and psychosomatic disorders and are known clinically to produce a sedation effect. Time-access alcohol-drinking behavior was determined at 24 hr after the termination of EAC. Finally, brain dopamine (DA) levels were assayed in the two groups. A sham-control group underwent only restricted stress without EAC. RESULTS: Time-access alcohol-drinking behavior increased significantly in the long-restricted group compared with the short-restricted group and controls. EAC applied to the ST 36 (Tsu-San-Li) point suppressed the increased alcohol drinking behavior in restricted rats. However, EAC applied to the Shen-Shu (BL 23) point was not effective, because alcohol-drinking behavior was significantly increased in long-restricted rats compared with short-restricted rats. Striatal DA levels of restricted rats with EAC stimulated at Tsu-San-Li were increased significantly compared with the rats with EAC applied to the Shen-Shu point. CONCLUSION: These findings suggest that EAC applied at ST 36 (Tsu-San-Li) was more effective for reducing the increased alcohol-drinking behavior in restricted rats, and they showed that a point specific in EAC procedure was associated with an increase of striatal DA levels. These findings provide new information for understanding alcohol-drinking behavior and for treating human alcoholics. PMID- 11410746 TI - A clinicopathological study of acute hepatitis in heavy drinkers, unrelated to hepatitis A, B, or C viruses. AB - BACKGROUND: There are six histological classifications of alcoholic liver disease (ALD) in Japan. However, it is unclear whether all cases of the disease conform to these criteria. This study investigated the clinicopathological features of eight histologically unusual cases of ALD. METHODS: The characteristic features of alcohol drinking behavior, subjective and objective symptoms, laboratory data on admission, and progress after admission were analyzed for eight patients with acute-onset hepatitis. RESULT: The eight patients showed histologically acute hepatitis, with much spotty necrosis that contained granular ceroid pigment by Kupffer cells, which indicated acute parenchymal damage of the liver, but with no Mallory bodies and unremarkable intrasinusoidal neutrophilic infiltration. The only etiological factor for all the cases was habitual alcohol consumption, with increased consumption just before the onset of symptoms. In five cases that were tested, the patients were negative for hepatic viral markers, which included hepatitis G virus RNA and TT virus DNA. CONCLUSION: Some cases of ALD may not conform to the current histological classifications in either Japan or Western countries. It seems natural to consider that these cases are developed by other, unknown causes that overlap with ALD rather than as a result of damage from alcoholic overload. PMID- 11410747 TI - Effect of fenofibrate on fatty liver in rats treated with alcohol. AB - BACKGROUND: Although fatty liver and hyperlipemia are common in chronic alcoholics, there is no practical approach to prevent alcoholic fatty liver. Recently, it has been reported that fibrates bind to peroxisome proliferator activated receptor-alpha and induce beta-oxidation enzymes of fatty acid in mitochondria. In this study, we investigated the effect of fenofibrate, one of the fibrates, on fatty liver in rats induced by chronic alcohol feeding. Furthermore, we studied the effect of fenofibrate on hyperlipemia in patients with alcoholic fatty liver. METHODS: Male Wistar rats were treated with liquid diet that contained ethanol (36% of total calories) or an isocaloric carbohydrate instead of ethanol for 4 weeks. Fenofibrate was administered orally with the liquid diets for 4 weeks at a concentration of either 0, 5, or 30 mg/kg body weight/day. As a pilot study, eight patients with alcoholic fatty liver were treated with 200 mg/day of fenofibrate for 4 weeks. RESULTS: After fenofibrate administration, fatty degeneration of liver was not observed in three of the five rats treated with 5 mg and in all rats treated with 30 mg of fenofibrate. Hepatic triglyceride content was decreased significantly in rats treated with 30 mg of fenofibrate compared with the rats not treated with fenofibrate. Serum triglyceride and total cholesterol levels also were decreased after treatment with fenofibrate. In eight alcoholic patients treated with 200 mg of fenofibrate for 4 weeks, serum triglyceride level decreased significantly compared with the levels before treatment. All patients continued alcohol consumption during fenofibrate administration. CONCLUSION: The results of the present investigation suggest that fenofibrate may be useful to prevent alcoholic fatty liver. Further studies with larger numbers of patients are necessary to obtain definitive results. PMID- 11410749 TI - Characterization of continuously extruded iopromide-carrying liposomes for computed tomography blood-pool imaging. AB - RATIONALE AND OBJECTIVES: Contrast-carrying liposomes are potentially useful as computed tomography (CT) blood-pool agents. In the present study, preliminary safety, pharmacokinetics, and the CT imaging behavior of continuously extruded iopromide-carrying liposomes were studied. METHODS: Iopromide liposomes were prepared by continuous high-pressure extrusion. Cell membrane-damaging characteristics were assessed in vitro in dog erythrocytes. Acute and subchronic toxicity and pharmacokinetics parameters were determined in rats. Computed tomography imaging efficiency was studied in rabbits. RESULTS: The iopromide carrying liposomes caused only minor morphological changes in dog erythrocytes. The median lethal dose in rats was approximately 4.5 g of total iodine per kilogram of body weight. In a subchronic tolerance study in rats that were administered six doses of 1 g iodine per kilogram twice a week, no adverse effects were observed. The pharmacokinetics in rats was dose dependent, and elimination of iopromide was almost complete within 7 days after intravenous administration. In rabbits, at a dose of 300 mg total iodine per kilogram, the iopromide-carrying liposomes displayed prolonged blood circulation, with mean CT density differences > 60 Hounsfield units (aorta) for up to 10 minutes. CONCLUSIONS: The iopromide liposomes were well tolerated, almost completely excreted, and have potential as a CT blood-pool imaging agent. PMID- 11410750 TI - Pharmacokinetics and safety of iomeprol in healthy volunteers and in patients with renal impairment or end-stage renal disease requiring hemodialysis. AB - RATIONALE AND OBJECTIVES: To present the results of two studies conducted to evaluate the pharmacokinetics and safety of iomeprol in healthy volunteers and in patients with various degrees of renal impairment. METHODS: In these two open label, single-dose, phase I studies, a 50-mL dose of iomeprol 400 was administered intravenously to a total of 30 subjects of either sex. In study 1, six healthy volunteers with normal renal function, six patients with mild renal failure, six patients with moderate renal failure, and four patients with severe renal failure were enrolled. In study 2, eight patients with end-stage renal disease requiring hemodialysis were enrolled. Safety was determined by predose and postdose (up to 10 days) measurement of vital signs, hematology, blood chemistry, urinalysis, electrocardiogram, physical examinations, and the incidence of adverse events. Pharmacokinetics was determined by measuring iomeprol levels in plasma, urine, feces, and dialysate samples, by using a validated high-performance liquid chromatography assay, up to 7 days after administration. RESULTS: The plasma concentration of iomeprol declined biexponentially in both healthy subjects and patients. As expected, mean body and renal clearances decreased progressively with increasing renal impairment, with a significant correlation with the glomerular filtration rate. The elimination half life increased progressively with increasing renal impairment. The extraction efficiency of dialyser was estimated as approximately 40%, and dialysis clearance of iomeprol was approximately 1.26 mL. min-1. kg-1 (80.6 mL/min), slightly less than the body clearance previously observed in healthy subjects. It appears that dialysis is almost as efficient as renal function in healthy subjects in the removal of iomeprol. After a single dialysis session, approximately 58% of the dose was recovered in dialysate. Mild to moderate adverse events were reported by 17 of 30 subjects; none was clinically meaningful. One serious adverse event, unrelated to iomeprol, was reported. No clinically meaningful findings were noted for other safety parameters. CONCLUSIONS: Iomeprol was almost completely eliminated both in patients with renal impairment and in patients receiving dialysis. No dose adjustment appears to be necessary either in patients with renal impairment or with end-stage renal disease requiring hemodialysis. In this risk population, iomeprol 400 was safe and well tolerated. PMID- 11410751 TI - Mechanical thrombectomy of major and massive pulmonary embolism with use of the Amplatz thrombectomy device. AB - RATIONALE AND OBJECTIVES: To evaluate the feasibility of mechanical thrombectomy with the Amplatz thrombectomy device (ATD) in restoring patency of acutely thrombosed pulmonary arteries resulting from pulmonary embolism for the improvement of patient outcome. METHODS: Mechanical thrombectomy with the ATD (8F) was performed in nine consecutive patients with angiographically documented thrombus in the left or right pulmonary artery resulting from deep vein thrombosis (n = 4) or unknown cause (n = 5). RESULTS: The Miller index decreased from 18 to 11. In all patients, the majority of the thrombus in the pulmonary artery was cleared after a mean activation time of the ATD of 367 seconds. Thrombectomy was performed with the ATD alone (n = 4) or with additional long term fibrinolysis therapy (n = 5) with infusion of recombinant tissue-type plasminogen activator. Pulmonary arterial pressure decreased from a mean of 57 mm Hg before mechanical thrombectomy to 55 mm Hg directly after the procedure and to 39 mm Hg after termination of the recombinant tissue-type plasminogen activator infusion. CONCLUSIONS: Mechanical thrombectomy with the ATD in patients with minor and major pulmonary embolism is technically feasible and safe. It is a potential alternative to drug-mediated thrombolysis and surgery. However, the incremental benefit of the ATD over conventional treatments could be shown only in a randomized controlled study. PMID- 11410752 TI - Do ultrasound measurements reflect bone microarchitecture rather than bone mass?: An in vitro study of the rat femur with the use of ultrasound, densitometry, and histomorphometry. AB - RATIONALE AND OBJECTIVES: In an experimental study in 40 rat femurs, the authors correlated the amplitude-dependent speed of bone ultrasound (Ad-SOS) with the bone mineral content and density and with the bone trabecular connectivity: trabecular bone volume, trabecular number, trabecular thickness, and trabecular separation to evaluate and compare the usefulness of the Ad-SOS to determine bone quantity and/or quality. METHODS: Bone mineral content and density were determined with dual-energy x-ray absorptiometry. Trabecular connectivity was determined with histomorphometric techniques. RESULTS: There was a strong correlation between the Ad-SOS and the other parameters studied, with a particularly high positive correlation with trabecular bone volume and trabecular thickness, and an inverse correlation with trabecular separation. The correlation was weaker with the bone mineral content and bone mineral density and with the trabecular number. For the trabecular separation, the correlation was significant in all cases, but it was negative. CONCLUSIONS: Bone ultrasound, in this case Ad SOS, defines the quality of the bone in terms of trabecular architecture rather than bone density; however, this conclusion is valid only for the rat femur model that the authors used. PMID- 11410753 TI - Ex vivo evaluation of novel miniaturized laser-induced interstitial thermotherapy applicators for effective small-volume tissue ablation. AB - RATIONALE AND OBJECTIVES: For effective small-volume tissue ablation in clinical and experimental settings, smaller laser-induced interstitial thermotherapy (LITT) applicator designs are required. The aim of this study was to compare the ablation properties of recently developed ultrasmall and small to standard LITT applicators. METHODS: Laser-induced interstitial thermotherapy was performed on liver samples using ultrasmall, small, and standard LITT applicators. Thermotherapy was monitored by magnetic resonance imaging, and lesion sizes were measured for each image. True lesion sizes were then determined macroscopically and by histology. RESULTS: For continuous laser application over 5 minutes, maximum power settings were 5 W for the ultrasmall and small applicators and 10 W for the standard applicator. Given identical LITT settings, lesion volume measured by magnetic resonance imaging was significantly larger and histological tissue damage was more severe with the ultrasmall and small applicators than with the standard applicator. CONCLUSIONS: Small and ultrasmall LITT applicators can be used for effective tissue ablation of small target volumes in experimental and clinical applications. PMID- 11410754 TI - Contribution of power Doppler sonography to the detection of renal allograft rejection in the cynomolgus monkey. AB - RATIONALE AND OBJECTIVES: To evaluate whether a change in the power Doppler (PD) flow signals produced by the renal cortical interlobular vasculature of allografts in cynomolgus monkey transplant models is useful for the detection of cellular rejection and vasculopathies. METHODS: Seventy-three monkeys with life supporting allografts (bilateral native kidney nephrectomy) and 20 monkeys with allografts implanted with only unilateral native kidney nephrectomy were examined with ultrasound that included an examination with PD. Each graft received a PD score of 3 (normal cortical blush), 2 (reduced flow, no blush), or 1 (absence of cortical flow), and the results were compared with histology either from ultrasound-guided biopsy or at necropsy. RESULTS: One hundred seventy-one allograft examinations (histological and PD) were compared. Histologically normal grafts were statistically more likely to have normal PD findings than were those with reduced flow or absent flow. Allografts with reduced flow had statistically more severe cellular rejection than those with normal flow. Also, vasculopathies were present in all three PD groups. CONCLUSIONS: Reduced renal cortical flow in the cynomolgus monkey renal allograft indicates that more severe degrees of cellular rejection are present compared with allografts with normal flow. Overlap in the histological diagnoses of allografts with normal and reduced flow exists, and the finding of reduced flow with PD may be prognostically important and indicates the need for tissue sampling. PMID- 11410755 TI - Short- and long-term repeatability of intrarenal resistive index in the pig. AB - RATIONALE AND OBJECTIVES: To evaluate the short- and long-term repeatability (intraobserver variation) of the renal resistive index (RI) in a pig model. METHODS: Under standardized ambient conditions, short- and long-term repeatabilities were assessed by measuring the RI three consecutive times at 30 minute intervals in 14 pigs and twice 4 weeks apart in 6 pigs, respectively. Repeatability was judged according to predefined criteria, which included calculation of coefficients of variation (CVs) and repeatability coefficients combined with visual assessment and estimation of bias. RESULTS: Short-term repeatability was acceptable, having an average CV of 4.9% +/- 2.9% and of 6.1% +/- 4.7% in two comparisons, findings that were reiterated in the visual assessment. Long-term results were, however, less consistent, having an average CV of 8.1% +/- 6.1% and prominent scatter on the visual assessment graphs. CONCLUSIONS: In this study, RI measurements showed an acceptable short-term repeatability, whereas long-term results were less consistent. The components of error contributing to the modest long-term repeatability need to be investigated in studies of larger populations. PMID- 11410756 TI - Displacement of breast tissue and needle deviations during stereotactic procedures. AB - RATIONALE AND OBJECTIVES: The aim of this study was to quantify the displacement of breast tissue and the inaccuracy of needle positioning for biopsy (14-gauge) and localization (19.5-gauge) needles. METHODS: For displacement of breast tissue, differences between the coordinates of identifiable microcalcifications in the images before (baseline) and after needle positioning were analyzed (n = 52). For accuracy of needle positioning, differences between the coordinates of the needle tip and the target were analyzed in breast tissue (n = 97) and in air (n = 246). RESULTS: Average target displacement was 2.1 mm for biopsy needles (95% prediction interval [PI] 0.6-7.8) and 1.0 mm (95% PI 0.3-3.9) for localization needles. Mean inaccuracy of needle positioning in breast tissue was 1.1 mm (95% PI 0.4-3.0) and 1.8 mm (95% PI 0.7-4.6) for biopsy and localization needles, respectively. CONCLUSIONS: Tissue and needle displacements cause a total positioning error of 2.4 mm in stereotactic core biopsy, which will limit the attainable diagnostic accuracy. PMID- 11410757 TI - Personality trait and disposition-a new opening for functional neuroimaging. PMID- 11410758 TI - Reply to Cheron and Dan. PMID- 11410760 TI - Arthritis in older adults. PMID- 11410761 TI - Treatment of rheumatic diseases in the elderly. PMID- 11410762 TI - Osteoarthritis overview. AB - Osteoarthritis (OA) is a nearly universal, slowly progressive degenerative condition affecting men and women as they age. OA of the hip and knee represent two of the most significant causes of pain and physical disability in adults. In the United States, OA is the second most common form of disability, and although it affects both men and women, women are more likely to be symptomatic. Despite its prevalence, OA is not necessarily an inevitable consequence of aging. Many advances have been made in understanding the pathophysiology and epidemiology of OA, and these advances are helping to dispel such attitudes. Evolving recognition of predisposing risk factors and pathophysiology suggest prudent steps that can be recommended. This article provides information for caretakers that will help them understand the disease process and plan appropriate therapeutic interventions. PMID- 11410763 TI - Gouty arthritis: not just a big toe problem. AB - Gout is a metabolic disease characterized by recurrent episodes of acute arthritis. High blood levels of uric acid lead to inflammation, joint swelling, and severe pain. Treatment is aimed at relieving the acute attack and preventing recurrent episodes of gouty arthritis. PMID- 11410764 TI - Self-help care in older African Americans with arthritis. AB - Arthritis is the third most common health problem among African Americans and the leading cause of activity limitations. Some of the obstacles facing people who are experiencing health care problems fall disproportionately on older racial and ethnic minority populations. Receiving care, which is perceived as racially and ethnically relevant, is an important factor in overcoming these obstacles. The Arthritis Self-Help Course (ASHC) was successfully used with 150 impoverished African-American elders living in public housing to bolster their confidence in managing their arthritis symptoms. PMID- 11410765 TI - Rheumatoid arthritis and the older adult. AB - Rheumatoid arthritis (RA) is the second most common rheumatic disease after osteoarthritis, but it is the most destructive to synovial joints. RA can be defined as a chronic, progressive, systemic, inflammatory disease of connective tissue characterized by spontaneous remissions and exacerbations (flare-ups). Unlike osteoarthritis, it involves primary tissue inflammation rather than joint degeneration. Although most individuals develop RA in early to middle adulthood, some experience a late onset RA (LORA) in their older years. The onset may be acute and severe or slow and insidious, making diagnosis difficult and often delayed. Regardless of the age at onset, RA affects many older adults in both community and inpatient settings and presents a challenge for nursing care. PMID- 11410766 TI - Managing arthritis with exercise. AB - Almost half of all older adults have arthritis, either degenerative or inflammatory. Regular exercise is an important therapeutic intervention for all types of arthritis. Specifically, regular exercise can prevent deconditioning of the muscles, keep the joints stable, improve joint function and flexibility, decrease pain, enhance aerobic fitness, improve balance, and decrease falls. A comprehensive exercise program should include stretching exercises followed by a range-of-motion program for joints, muscle strengthening, and aerobic exercise, if possible. Unfortunately, despite these known benefits, most older adults with arthritis do not engage in regular exercise. The Seven Step Approach was developed to provide a practical framework to help overcome barriers and improve exercise activity in older adults with arthritis. PMID- 11410767 TI - Assessment and management of polymyalgia rheumatica in older adults. AB - Polymyalgia rheumatica (PMR) is a periarticular rheumatic condition characterized by pain and stiffness, primarily in the neck, shoulders, hips, and pelvic girdle. Temporal arteritis (TA) or giant cell arteritis, the most common primary vasculitis in older adults, is found in approximately 10% to 30% of people who have PMR. Left untreated, TA can result in sudden, irreversible blindness. Geriatric nurses need to familiarize themselves with these disorders to accurately assess and manage people with them. PMID- 11410768 TI - Joan Warden-Saunders. PMID- 11410769 TI - Challenges in the care of adults with osteoporosis. PMID- 11410770 TI - Newer and safer options for osteoarthritis. PMID- 11410771 TI - Meatless diets part 1. PMID- 11410772 TI - Radical cystectomy for invasive bladder cancer in the octogenarian. AB - The incidence of bladder cancer increases with age. As the population lives longer, an increasing number of patients 80 years of age or older will develop invasive bladder cancer. In this study, we reviewed the outcome of 33 patients age 80 years or older treated with radical cystectomy and ileal conduit urinary diversion. Five patients received neoadjuvant chemotherapy, and 2 had salvage cystectomy after failure of external beam radiation therapy. The median age was 82 years, and the median hospital stay was 12 days. There were no perioperative deaths. Twenty-seven complications occurred in 20 patients (60.6%), of which 17 were minor (63%) and 10 were major (37%). There was no difference in the rate of complications in patients receiving neoadjuvant treatment compared to the group treated with cystectomy alone. The median survival was 3.5 years. Our results demonstrate that radical cystectomy and ileal conduit urinary diversion should not be withheld from patients on the basis of age. PMID- 11410773 TI - Absence of therapeutically relevant c-erbB-2 expression in human hepatocellular carcinomas. AB - c-erbB-2 (Her-2/neu) represents a well established prognostic marker and therapeutic target in several human tumor types, especially breast cancer. Its potential role in hepatocellular carcinoma (HCC) has not yet been characterized. Using standardized immunohistochemical procedures, we have analyzed the c-erbB-2 expression in a representative panel of human HCCs. Our data demonstrate the absence of significant membranous expression or cytoplasmic overexpression. Thus c-erbB-2 is neither a prognostic marker nor a relevant therapeutic target in human HCCs. PMID- 11410774 TI - Effect of giant hepatomas on lymphocyte production and secretion of apoptosis related proteins in the rat spleen. AB - The effect of extremely large hepatomas on splenic lymphoid elements and apoptosis-related proteins in rats were studied. Hepatoma cells were inoculated subcutaneously into 6-week-old rats, and 4 months later the quantities of T and B cells, macrophages, and cells positive for Fas, Fas ligand (FasL) and interleukin 2 (IL-2) were immunohistochemically evaluated in spleens. Grafting of hepatoma cells caused hyperplasia of the spleen and development of giant tumors that could reach one-third of the rat's body weight. A 7-fold increase in the weight of the spleen was mainly due to proliferation of B lymphocytes and macrophages in the red pulp, while the relative quantity of CD4+ and CD8+ T cells decreased. Extremely small amount of Fas+ and FasL+ lymphocytes were present in the marginal zone, the follicles, red pulp, and occasionally in the PALS. All the splenic zones were abundant with IL-2+ cells, while macrophages and siderophages were present mainly in the red pulp and in the marginal zone of the white pulp. We suggest that all these changes are compensatory processes of the host's lymphatic system. PMID- 11410775 TI - Prognostic significance of p21waf1, cyclin D1 and retinoblastoma expression detected by immunohistochemistry in non-small cell lung cancer. AB - p21waf1 is a downstream effector of p53, and mediates growth arrest by inhibiting the action of G1 cyclin-dependent kinases. Cyclin D1 is a cell-cycle regulator essential for G1 phases progression and a candidate proto-oncogene implicated in the pathogenesis of several human tumor types. Cyclin D1 overexpression and the absence of retinoblastoma (Rb) protein have been frequently seen in various types of cancer, including lung cancer. The aim of this study was to clarify the relationship between the expressions of p21waf1, cyclin D1, and Rb protein, and to investigate the correlation between these protein expressions and the clinical features of the patients, and their prognoses. We immunohistochemically examined 92 samples of resected non-small cell lung cancer for p21waf1, cyclin D1, and Rb expression. Of the 92 specimens examined, 43 cases (46.7%) showed p21waf1 expression, 23 cases (25.0%) showed cyclin D1 overexpression, and 61 cases (66.3%) showed Rb expression. No correlation was observed between the expressions of p21waf1, cyclin D1, and Rb. There was no association of p21waf1 and cyclin D1 immunoreactivity with gender, disease stage, or histological types of the tumor. Regarding the prognosis in 79 cases with complete resection, no statistical differences were observed according to the degree of expression of these three factors. However, when unfavorable prognostic factors were considered to be the positive expression of p21waf1, positive of cyclin D1, and negative of Rb, the 5 year disease-free survival rate in the group with 2 or 3 unfavorable prognostic factors was 21.1%, which was statistically poorer than the 45.4% in the group with 0 or 1 unfavorable prognostic factor (p=0.0138). We conclude that examination of the expression of cell cycle regulators, such as p21waf1, cyclin D1, and Rb, is useful as a prognostic indicator, when these proteins' expression is analyzed in combination. PMID- 11410776 TI - Phenotypical characteristics of undifferentiated carcinoma of the pancreas: a comparison with pancreatic ductal adenocarcinoma and relevance of E-cadherin, alpha catenin and beta catenin expression. AB - The characteristics of undifferentiated (anaplastic) carcinoma (UC) of the pancreas were analysed and compared with those of poorly differentiated adenocarcinoma (PA). Eight cases of UC of the pancreas were evaluated by clinicopathological and immunohistochemical methods and compared with 20 cases of PA. Large size of the tumor, local invasion and lymph node metastasis were frequently seen in UC, leading to a significantly worse prognosis compared to that of PA (p=0.022). Immunohistochemically, E-cadherin expression was completely lost in 7/8 UC cases, whereas half the PA cases revealed a strong reactivity for E-cadherin. alpha- and beta-catenin expressions tended to be impaired in UC compared with PA. The results suggest that the altered expression of adhesion molecules is correlated with dedifferentiated change and is contributory to its aggressive biological behaviour. PMID- 11410777 TI - Thymidine phosphorylase expression and effect of doxifluridine: a phase II study. AB - Doxifluridine (5'-DFUR), an active intermediate metabolite of capecitabine, is converted to 5-fluorouracil by thymidine phosphorylase (TP). We used immunohistochemical staining to investigate the relation between TP expression and 5'-DFUR effects in 40 patients with advanced/recurrent lung metastases from colorectal cancer. Cox regression analysis suggested that TP-positive cancer cells (risk ratio 3.72), were independent factors in survival whereas factors in progression-free survival were TP-positive cancer cells (2.93), and TP-positive stromal cells (0.24). It is suggested that TP expression in cancer cells and in stromal cells are opposite prognostic factors in patients treated with 5'-DFUR. PMID- 11410778 TI - Expression of thymidylate synthase in cancer of the ampulla of Vater. AB - The clinical and therapeutic significance of thymidylate synthase (TS) in cancers of the ampulla of Vater have not yet been reported. We immunohistochemically evaluated TS expression in 33 ampullary cancers using an anti-TS antibody. TS expression, clinicopathologic variables, and survival rates were examined and the correlations between these parameters were identified. Fifteen patients were found to express high levels of TS (high TS group), while 18 patients expressed low levels of TS (low TS group). No significant difference was found between TS expression and clinicopathologic factors. Univariate and multivariate analysis revealed that lymph node metastasis and pancreatic invasion are important variables for independently predicting post-operative survival. Although TS expression was not identified as an important factor for postoperative survival, recurrent cases in patients with chemotherapy existed only in the high TS group. In the present study, it was found that TS expression itself in cancers of the ampulla of Vater has no impact in predicting the prognosis of ampullary cancers, but a chemotherapeutic benefit of evaluating TS expression may exist. PMID- 11410779 TI - Expression of thrombospondin-1 inversely correlated with tumor vascularity and hematogenous metastasis in colon cancer. AB - Solid tumors require neovascularization for growth and metastasis. Angiogenesis depends on the local balance between various molecules that induce and inhibit neovascularization. Thrombospondin-1 (TSP-1) is thought to be an antiangiogenic factor. In this study, we examined the correlation between expression of TSP-1 and tumor vascularity, and determined its prognostic significance in colon cancer. Microvessel density, determined by immunostaining for factor VIII-related antigen, was significantly higher in tumors that were TSP-1-negative. TSP-1 expression was inversely correlated with prognosis. Patients with TSP-1-negative tumors had a significantly worse prognosis than did those with TSP-1-positive tumors. Frequency of hepatic recurrence was significantly higher in patients with tumors that were TSP-1-negative. In conclusion, TSP-1 is an important negative regulator of tumor angiogenesis, and TSP-1 may be useful for predicting recurrence in patients with colon cancer. PMID- 11410780 TI - Effect of human urinary macrophage colony-stimulating factor on the immune functions and the blood cell counts in patients with ovarian carcinoma receiving cytotoxic chemotherapy. AB - The present study investigated the effect of macrophage colony-stimulating factor (M-CSF) on the immune functions and blood cell counts of patients with ovarian carcinoma receiving cytotoxic chemotherapy (CTX). Seventy-five consecutive patients with white blood cell counts less than 3,000/microl after CTX were randomly assigned to receive either M-CSF (human urinary macrophage colony stimulating factor: hM-CSF, 8x106 U as 7-day intravenous infusions) or no treatment. Immune assays in addition to routine peripheral blood examinations were performed on these patients at various time points. hM-CSF dosing significantly increased monocyte, lymphocyte, granulocyte, and platelet counts that were decreased by CTX. hM-CSF also significantly enhanced lymphokine activated killer and natural killer activities, which was accompanied by a significantly augmented interleukin (IL)-2 production. Interestingly, IL-2 production was enhanced by hM-CSF dosing in 24 of the 27 patients with a pre-hM CSF level of IL-2 below 10 U/ml, but such an effect was not observed in nine of the 10 patients having a basal value of 10 U/ml or higher. Thus, hM-CSF is considered to be a cytokine that can augment or regulate immune functions impaired by CTX and increase blood cell counts that are decreased by CTX. PMID- 11410781 TI - Combined diagnostic imaging for retroperitoneal schwannoma. AB - Retroperitoneal tumors are occasionally encountered by gynecologists, and differential diagnosis from adnexal tumors is important, however, it is difficult in many cases. We report a case in which we preoperatively diagnosed a benign solid tumor in the retroperitoneum of the pelvic cavity on the basis of a variety of diagnostic images, including FDG-PET. Postoperative histological examination showed a benign schwannoma in the pelvis. We discuss the usefulness of FDG-PET combined with MRI and other diagnostic images as a means of differentiating between the characteristics of retroperitoneal tumors. PMID- 11410782 TI - Excellent results of radiotherapy for neuroendocrine carcinoma of the uterine cervix. AB - Neuroendocrine carcinoma of the uterine cervix is a rare neoplasm that is associated with a poor prognosis, and there have been no reports of excellent results of radiotherapy to treat it. We report a case of neuroendocrine uterine cervical carcinoma, as stage IIIa (FIGO), in which the patient showed a complete clinical response to radiotherapy. PMID- 11410783 TI - Patterns of brain metastasis in lung cancer. AB - Brain metastasis as the only site of spread in lung cancer was studied. Ninety two (13.1%) of 701 lung cancer patients had brain metastasis at the time of diagnosis. Eighteen and 16 patients had small primary lesion and were N0-1 disease, respectively. Thirty-two patients had sole brain metastasis. Patients with sole brain metastasis had more favorable survival than patients with other metastatic sites (p=0.0165). Brain metastasis may occur even with small primary lesion and/or without regional lymph node involvement. Well-planned surgery or stereotactic radiosurgery will result in enhanced survival. PMID- 11410784 TI - A pilot study of radiation therapy combined with daily low-dose cisplatin for esophageal cancer. AB - From March 1992 to October 1997, a total of 38 patients with histologically proven esophageal cancer received combination therapy of daily low-dose cisplatin (CDDP) and radiation therapy. The clinical stages (UICC 1997) were I in 3, IIA,B in 13, III in 13, and IVA in 9 patients. CDDP (5 mg/m2) was administered intravenously with 100 ml saline 30 min before each irradiation from Monday to Friday. All patients received at least 60 Gy of radiation therapy. The average number of CDDP administrations was 23, and the mean total CDDP dose was 115 mg/m2. Of the 38 patients, 14 patients completed full course of chemoradiation therapy. The overall survival rates at 1, 2, and 5 years were 51%, 19%, and 8%, respectively. The median survival period was 12 months. The objective response rate was 82% with 11 (29%) CR. Survival of the stage II-IVA patients who received daily low-dose CDDP and radiation therapy was a little better than that of historical control patients who were treated by radiation therapy alone. Most of the patients experienced nausea. Grade 3 esophagitis was observed in two (5%) patients. Grade 4 leukocytopenia and thorobocytopenia were observed in one (3%) and two (5%) of the patients, respectively. Except for leukocytopenia (18%), frequencies of toxicity of grade 3 or more were less than 10%. Although the results indicate that daily low-dose CDDP combined with radiation therapy may slightly improve the survival of esophageal cancer patients with acceptable toxicity, further efforts should be made to optimize clinical trial protocols. Escalation of daily CDDP dose should be considered to obtain a more effective radiosensitizing effect. PMID- 11410785 TI - No mutations of the Bub1 gene in human gastric carcinomas. AB - Chromosomal instability in colorectal cancers is associated with functional loss of a mitotic check point partly due to mutations of the Bub1, one of the mitotic check point genes. However, mutation of coding sequences of human Bub1 gene has not been fully elucidated in gastric carcinomas. We performed sequencing analysis on reverse transcriptase-polymerase chain reaction (RT-PCR) product of the Bub1 cDNA (entire coding sequence) from 5 human gastric carcinomas as well as on genomic PCR products of Bub1 kinase domain from 7 gastric carcinoma tissues. Although sequencing analysis of the Bub1 cDNA revealed several point mutations in 2 gastric carcinoma cases, we could not confirm the mutations by analyzing genomic DNA. Furthermore, genomic DNA sequencing revealed no mutations in the kinase domain of the Bub1 gene in any gastric carcinoma examined. These results suggest that mutational inactivation of the Bub1 gene might not play a key role in human stomach carcinogenesis. PMID- 11410786 TI - Development of terminal cancer prognostic score as an index in terminally ill cancer patients. AB - Cancer patients, families, clinicians, and health-policy administrators need accurate information about the prognosis of survival of terminally ill cancer patients. The aim of this study was to compare survival times and prognostic factors, and develop a new prognostic index for terminally ill cancer patients. This prospective study was performed on 91 patients with solid tumor, and therefore, no longer subjects of anti-cancer therapy. Association was sought between survival times and a range of clinical characteristics. The median survival time of 91 terminal cancer patients was 54.0 days. Univariate analysis showed that 11 factors provide statistically significant prognostic survival information. Multivariate analysis adjusted for the primary tumor site demonstrated that severe anorexia (aRR 1.95, 95% C.I. 1.24-3.05), severe diarrhea (aRR 3.49, 95% C.I. 1.10-11.05), and mild confusion (aRR 1.94, 95% C.I. 1.15 3.27) are independent negative predictors of survival. The Terminal Cancer Prognostic score (TCP score), which was based on three predictors proved to be a significant predictor. The TCP score might be a useful index for predicting survival. PMID- 11410787 TI - Sequential docetaxel and vinorelbine for patients with advanced breast cancer previously treated with anthracyclines: a phase II study. AB - With respect to their association, sequential non-cross-resistant cytostatics could be better tolerated and allow a similar antitumor effect. From January, 1998 to July, 1999, 42 consecutive patients with metastatic breast cancer (MBC) previously treated with anthracyclines as adjuvant- or first-line therapy entered a phase II multicenter study where docetaxel (TXT, 100 mg/m2/3 weeks/4 times) was followed by vinorelbine (VNR, 25 mg/m2/10 days/8 times). Median follow-up is 21 months and 22/42 patients have died. Four patients did not complete therapy due to early death, grade 3-4 gastrointestinal mucosytis (2 patients) and grade 3 neurotoxicity during TXT therapy. Overall response rate was 57%, and 5% of patients had stable disease. There were 38% of therapy failures due to non evaluability (10%) or progressive disease (28%). Median time to progression and survival are 10.1 and 17.1 months. Sequential TXT-VNB is a suitable strategy for MBC patients previously treated with anthracyclines. It avoids haematologic toxicity and allows a good antitumor effect. Careful monitoring of intestinal mucosytis is required. PMID- 11410788 TI - Use of c-myb antisense oligonucleotides to increase the sensitivity of human colon cancer cells to cisplatin. AB - Human colon cancer SW480DDP and SW620DDP cells resistant to cisplatin exhibited stronger c-myb gene expression than the parent SW480 and SW620 cells. However, cell growth rates in the cisplatin-resistant cell lines remained similar to those of the parent cells. Antisense oligonucleotides to c-myb inhibited c-myb expression and induced increased sensitivity to cisplatin in SW480DDP and SW620DDP cells, but this did not occur with the control sense oligonucleotides. In contrast, the parent cell lines were not affected by antisense oligonucleotides to c-myb. These results indicate that the c-myb gene in human colon cancer is one of the factors related to cisplatin resistance, and support the need to develop anti-cancer therapeutics based on oncogene-targeted antisense oligonucleotide technology. PMID- 11410789 TI - A case of pancreatic serous cystadenoma obstructing the distal pancreatic duct. AB - We present a case of resected serous cystadenoma of the pancreas inducing marked dilatation of the main distal pancreatic duct. A 68-year-old woman, previously diagnosed with chronic pancreatitis, presented with upper abdominal pain. Abdominal US revealed a highly echoic mass in the pancreas. A CT scan disclosed a low density mass in the pancreas and dilatation of the main peripheral pancreatic duct. The mass demonstrated homogeneous and high signal intensity on T2-weighted magnetic resonance imaging (MRI). Selective abdominal arteriography showed the mass strained by the celiac artery. The tumor markers were CEA (2.4 ng/ml) and CA19-9 (6.1 U/ml). After the diagnosis of serous cystadenoma of the pancreas, the patient underwent distal pancreatectomy and splenectomy. The tumor (2.5 cm in diameter) consisted of grayish-white nodules and occupied the body of the pancreas. The tail of the pancreas was atrophic. Histopathological examination of the specimen showed a multilocular lesion containing numerous cysts with the inner surfaces evenly lined by one layer of cuboid or flat epithelial cells which stained positive for periodic acid-Schiff (PAS), evidencing serous cystadenoma. The patient is doing quite well one and a half years after the operation. PMID- 11410790 TI - Effects of MDR1/P-glycoprotein expression on prognosis in advanced colorectal cancer after surgery. AB - Resistance to chemotherapeutic agents is a major problem for successful cancer treatment. P-glycoprotein (Pgp), a product of the multidrug resistance (MDR)1 gene expressed in cancer cells, is one of the mechanism of MDR. However, there are few reports regarding the effects of Pgp on prognosis of colorectal cancer (CRC) after surgery. We examined a total of 80 patients (45 males and 35 females with an average age of 69 years) whose CRCs were classified into stage 2-4 and completely resected surgically in our institute between January 1990 and September 1999. To evaluate Pgp expression in CRC, immunohistochemical stain was performed with a monoclonal antibody. Relationships between Pgp expression and clinicopathological variables which may have affected prognosis were evaluated. Survival curves were calculated using the Kaplan-Meier method, and differences were evaluated with the log-rank test. The Cox's proportional hazards model was used in the univariate and multivariate survival analysis. Pgp expression showed a significant correlation with histological differentiation (p=0.023). However, no correlation was observed with gender, tumor location, lymph node metastasis, lymphatic invasion, venous invasion, and cancer stages. Survival rates after surgery tended (p=0.093) to be higher in Pgp (+) than Pgp (-) patients. Pgp was not a significant prognostic factor by univariate analysis and multivariate analysis adjusted for other clinicopathologic variables. Survival rates after surgery tended to be higher in Pgp (+) than Pgp (-) patients and Pgp expression was correlated with histological differentiation of CRC. Thus, a relative resistance of CRC to conventional chemotherapy may be partly caused by Pgp expressed in well or moderately differentiated CRC. However, Pgp expression was not a significant independent prognostic factor in advanced CRC after surgery. PMID- 11410791 TI - Cycloprodigiosin hydrochloride, a H+/Cl- symporter, induces apoptosis in human colon cancer cell lines in vitro. AB - Recently, we reported that cycloprodigiosin hydrochloride (cPrG.HCl), a novel H+/Cl- symporter, induces acidification of the cytosol and leads to apoptosis on rat and human liver cancer cells. In the present study, the effects of cPrG.HCl, a H+/Cl- symporter, were examined in colon cancer cell lines in vitro. In the MTT assay, cPrG.HCl inhibited the growth of two colon cancer cell lines (WiDr and SW480) in a dose- and time-dependent manner. The cPrG.HCl treatment of both types of cells induced apoptosis as confirmed by the appearance of a sub-G1 population and intranucleosomal DNA fragmentation. In addition, cPrG.HCl lowered pHi (below pH 6.8) respectively. Therefore, these results suggest that cPrG.HCl may be useful for the treatment of colon cancer cells. PMID- 11410792 TI - Proteasome inhibitor 1 enhances paclitaxel-induced apoptosis in human lung adenocarcinoma cell line. AB - Paclitaxel is a chemotherapeutic drug that induces apoptosis in tumor cells by stabilizing microtubules, prevents normal mitosis, and blocks the cell cycle at the G2/M phase. We have previously reported that the activation of caspase-3 and caspase-8 plays a crucial role in paclitaxel-induced apoptosis. Anti-tumor reagents including paclitaxel, irradiation, and other stimuli activate the transcription factor NF-kappaB, which has the ability to suppress the apoptotic potential of those stimuli. Using a human lung adenocarcinoma cell line (LC-2 AD), we therefore examined whether the inhibition of NF-kappaB activity by proteasome inhibitor 1 (PS1) could become a new adjuvant therapy for cancer. A synergistic effect on apoptosis induction was observed with the combination of more than 0.1 microg/ml paclitaxel and 0.5 microM PS1. An increase in the cell number of apoptotic cells is correlated with the loss of Deltaphim and the activation of caspase-3 and caspase-8. Furthermore, augmented apoptosis is related to NF-kappaB activation. Based on these findings, we propose that the combination of paclitaxel with PS1 could be a new strategy for cancer treatment. PMID- 11410793 TI - Relationship of p53 and Helicobacter pylori to clinicopathological features of human remnant stomach cancer after gastric surgery for primary gastric cancer. AB - The purpose of this study was to evaluate the biological features of gastric cancer of the remnant stomach (RSC). Twenty-one patients underwent resection of the remnant stomach for RSC and were divided into two groups: the RSCB group consisted of 11 patients who underwent distal gastrectomy for benign disease and the RSCM group consisted of 10 patients who underwent gastrectomy for primary gastric cancer. The interval between primary surgery and the appearance of gastric cancer in the remnant stomach was significantly shorter in the RSCM group than in the RSCB group. Invasion of adjacent organs was more frequent in the RSCM group than in the RSCB group and the Ki-67 labeling index of the tumors was significantly higher in the former group. Furthermore, p53 overexpression by tumors was almost twice as common in the RSCM group as in the RSCB group. Although there was no significant difference of the H. pylori positivity between the two groups, the rate for both groups was higher than reported in previous studies. Mutation of p53 may play an important role in the high proliferative activity of tumors in the RSCM group and H. pylori infection may be closely related to carcinogenesis in patients with RSC. PMID- 11410794 TI - Breast-conserving therapy for ductal carcinoma in situ. AB - This retrospective analysis evaluates the treatment results and prognostic factors of 114 patients with ductal carcinoma in situ (DCIS) undergoing breast conserving therapy (BCT) at Keio University Hospital Department of Radiology, between 1988 and 1997. A total of 132 patients with DCIS of the breast came to our hospital between 1988 and 1997, and 114 cases were suitable candidates for BCT. All of the patients were female and ranged in age from 26 to 81 years (median 46). Ninety-one patients were premenopausal, and 23 were postmenopausal. Median clinical tumor size was 2.0 cm (0-8.0 cm). Postoperatively 48 cases received 50 Gy/25 fractions of external irradiation to the whole breast via tangential ports. The follow-up period after treatment ranged from 11 to 162 months (median 46.7). The local relapse-free rate and overall survival rate of the 114 patients were 89.5% and 100%, respectively. Local failure and regional nodal failure occurred in 12 and 1 patient, respectively. Radiotherapy was a significant risk factor for local failure (p=0.05). No postmenopausal patients developed local failure, but the difference did not reach statistical significance (p=0.103). The 12 recurrent cases underwent additional surgery and all remain alive without recurrence, to date, i.e., at least 16 months. Breast conserving surgery plus irradiation is appropriate treatment for DCIS patients. PMID- 11410795 TI - Clinical role of Tc-99m-MIBI scintigraphy in non-Hodgkin's lymphoma. AB - This study was designed to investigate the relationship between Tc-99m-hexakis-2 methoxyisobutylisonitrile (MIBI) scintigraphy and outcome of treatment in patients with non-Hodgkin's lymphoma (NHL). Forty-five patients with NHL were studied with Tc-99m-MIBI before any treatment. Images of the lesions were obtained at 20 min and 2 h after radionuclide administration. Visual semi quantitative interpretation was performed for Tc-99m-MIBI (grade 0-4) scintigraphy. Patients underwent 3-5 cycles of CHOP chemotherapy with/without involved field radiotherapy for large tumors. Their responses to treatment were evaluated at the end of chemotherapy and during the follow-up period. Forty of 45 patients (89%) showed abnormal uptake of Tc-99m-MIBI. There was no correlation between intensity of MIBI accumulation and response to chemotherapy. However, patients with negative or decreased MIBI activity 2 h after radionuclide administration showed worse response to chemotherapy compared to those with continued MIBI activity. MIBI activity could not predict the development of relapse in the follow-up study. In this study, the number of patients was small and we could not reach definite conclusions. However, we think that MIBI scintigraphy is not valuable for predicting the chemotherapy outcome in patients with NHL. PMID- 11410796 TI - Role of nitric oxide in human gastric cancer cells treated with 5-fluorouracil. AB - We examined whether 5-fluorouracil (5-FU) induces nitric oxide (NO) production and evaluated the role of NO in antitumor activity in human gastric cancer cells. MKN-1 gastric cancer cells were treated with the IC50 of 5-FU in the presence of interferon-gamma (IFN-gamma). In addition, s-methylisothiourea (an antagonist of inducible nitric oxide synthase) or anti-TNF-alpha antibody was added to the culture medium. Production of NO was measured by nitrite assay, TNF-alpha was measured by enzyme-linked immunoabsorbent assay, antitumor activity was evaluated by 3-[4,5-dimethylethiazol-2-yl]-2,5-dipheniltetrasolium bromide (MTT) assay. After 5-FU treatment in the presence of IFN-gamma, NO and TNF-alpha were produced and anti-tumor activity was enhanced. In contrast, s-methylisothiourea abolished the antitumor activity of 5-FU treatment. Anti-TNF-alpha antibody inhibited NO production and decreased the antitumor activity. 5-FU induces NO production by gastric cancer cells, and NO participates in antitumor activity in gastric cancer cells. These effects may be mediated by TNF-alpha production. PMID- 11410797 TI - Lung cancer in middle-aged patients. AB - Clinicopathological features of middle-aged lung cancer patients were investigated in 1016 consecutive patients. Among them, 22.8% of patients were in their forties and fifties. The preponderance of adenocarcinomas and the higher female/male ratio in middle-aged groups compared with the older group were findings similar to those observed in younger patients. Smoking habit increased according to the age groups. Middle-aged patients had a propensity for advanced stage, however, survival was not inferior to younger patients. Middle-aged patients consisted of two different groups of patients, whose characteristics were similar to those observed in either younger or elderly patients. PMID- 11410798 TI - Correlation between EGF receptor expression and peplomycin cytotoxicity in squamous cell carcinoma cell lines. AB - The relationship between sensitivity to anti-cancer agents and EGF receptor expression on squamous cell carcinoma (SCC) cells was investigated. The cytotoxicity of peplomycin (PEP) was correlated with the number of the EGF receptors expressed on the cancer cells, but no correlations were found between the cytotoxicity of adriamycin and cisplatin and EGF receptors. Addition of TNFalpha increased the number of EGF receptors in the SCC cell lines 1.5- to 2 fold. The cytotoxic effect of combined administration of PEP and TNFalpha was correlated with the number of EGF receptors, and produced a 2- to 5-fold increase in IC50 compared with administration of PEP alone. These observations suggest that EGF receptor expression is closely associated with the cytotoxic effect of PEP on SCC cells. PMID- 11410799 TI - Randomized study of dose or schedule modification of granulocyte colony stimulating factor in platinum-based chemotherapy for elderly patients with lung cancer. AB - It is generally believed that elderly patients are less able to tolerate aggressive cancer chemotherapy than their younger counterparts. Bone marrow cellularity diminishes with age and elderly patients may have decreased tolerance to myelosuppressive agents. Between November 1995 and October 1999, 68 chemotherapy-naive elderly (70 or more years old) patients with histologically or cytologically proven lung cancer who were to receive platinum-based chemotherapy were enrolled in this study. All patients had adequate cardiac, hematological, liver and renal function to receive chemotherapy. Patients were randomized into 3 groups. Patients in groups 1 and 2 received 2 microg/kg and 4 microg/kg granulocyte colony-stimulating factor (G-CSF, lenograstim), respectively, when grade 3 leukopenia (<2,000/microl) or neutropenia (<1,000/microl) appeared after chemotherapy. Patients in group 3 received 2 microg/kg G-CSF when grade 2 leukopenia (<3,000/microl) or neutropenia (<1,500/microl) appeared after chemotherapy. G-CSF was stopped in all groups when the leukocyte count increased to over 10,000/microl or the neutrophil count exceeded 5,000/microl. Full blood cell counts were examined 3 times a week after chemotherapy. All patients received platinum-based chemotherapy. Eighteen, 16 and 22 patients (78%, 73% and 96%) in groups 1, 2 and 3, respectively, received G-CSF when leukopenia or neutropenia appeared. The durations of G-CSF treatment required by groups 1 and 3 (5.7+/-3.6 and 6.6+/-3.2 days, respectively) did not differ significantly, but the duration of treatment required by group 2 (3.7+/-2.8 days) was significantly shorter than that of group 1 (p=0.048). The duration of grade 4 neutropenia in group 2 (0.7+/-1.1 days) was marginally shorter than that in group 1 (1.6+/-2.1 days, p=0.076). The neutrophil nadir of group 2 (949+/-757/microl) was marginally higher than that of group 1 (592+/-438/microl, p=0.058). No patients in group 2 experienced grade 4 neutropenia for 4 days or more or a neutrophil nadir less than 100/microl a significant difference from group 1, where 22% and 17% of patients experienced these events (p=0.02 and p=0.04, respectively). Similarly, no infections requiring antibiotics after chemotherapy occurred in patients in group 2, a significant difference from group 1 (26%, p=0.01). The rates of neutropenia and infection in groups 1 and 3 did not differ significantly. The peak plasma concentration of G-CSF in group 2 was significantly higher than in group 1 (p=0.0018), but did not differ significantly between groups 1 and 3. Doubling the dose of G-CSF could help to decrease neutropenia and prevent infection after chemotherapy in elderly patients with lung cancer. PMID- 11410800 TI - The usefulness and limits of magnetic resonance imaging in the differential diagnosis of pelvic tumors. AB - Three cases of benign pelvic tumors are presented (2 leiomyomas and 1 fibroma). All three tumors were suspected of being malignant neoplasms because they were visualized as heterogeneous high signal intensity on T2-weighted images, and thus they were difficult to diagnose preoperatively. One of the leiomyomas was located in the retroperitoneum and had been misdiagnosed as an ovarian tumor. All three tumors exhibited secondary myxoid changes, these changes may have been responsible for the high signal intensity on the T2-weighted MR images. Since benign tumors sometimes mimic malignant tumors on MR images, exploratory laparotomy is essential to make a definitive diagnosis. PMID- 11410801 TI - Increased expression of platelet-derived endothelial cell growth factor in human hepatocellular carcinomas correlated with high Edmondson grades and portal vein tumor thrombosis. AB - Platelet-derived endothelial cell growth factor (PD-ECGF), identical to thymidine phosphorylase, has been reported as an angiogenic factor in human malignancies. However, the role of PD-ECGF in human hepatocellular carcinoma (HCC) is still unconfirmed. Herein, we studied the expression of PD-ECGF in 27 human HCC cases by immunohistochemistry, to clarify the relationship to tumor angiogenesis. The immunoreaction of PD-ECGF in HCC cells was scored in both the staining percentage and intensity. CD34, an endothelial cell marker, was used to evaluate the intratumoral microvessel density (IMVD). PD-ECGF expression was noted in carcinoma cells in 14 (51.9%) of 27 HCCs. In these cases, the carcinoma cells showed heterogeneous staining in both the nucleus and cytoplasm. Tumor-associated stroma cells and infiltrating lymphocytes were also stained. Kupffer cells in non tumor areas were strongly positive. Statistically, the expression of PD-ECGF increased in HCC specimens with high Edmondson grades (III-IV) or portal vein tumor thrombosis (PVTT) (P<0.05). Additionally, the IMVD of PD-ECGF-positive HCC specimens (136.071+/-31.008, mean +/- SD) was higher than that of the PD-ECGF negative HCC specimens (61.077+/-15.795) (P<0.05). These findings may suggest that PD-ECGF is one of the angiogenic factors in human HCCs. Furthermore, with the increasing expression of PD-ECGF, HCC cells show poor differentiation and invasive behavior. PMID- 11410802 TI - Bone reaction induced by femoral stem of titanium alloy endoprosthesis for malignant bone tumors at the distal femur. AB - We demonstrated radiological evaluations after limb salvage surgery with an uncemented titanium endoprosthesis possessing a rotating hinge-type knee joint for 7 patients with malignant bone tumors at the distal femur. Although radiographically, cortical hypertrophy and new bone formation at the junction between implant and cortex appeared in all cases, there was a small quantity of subsidence of the implant in all cases. Loosening of the stem was found in 2 patients at 4 and 9 years after surgery. However, after revision surgery, the function restored as before. The latest average functional score according to the system of the International Society of Limb Salvage was good, being 73%. PMID- 11410803 TI - Relationship between dose-intensity of treatment and outcome for patients with osteosarcoma of the extremity treated with neoadjuvant chemotherapy. AB - One hundred and forty-four patients with osteosarcoma of the extremity treated with neoadjuvant chemotherapy at the authors' institution between 1986 and 1989 were retrospectively analyzed to evaluate the relationship between the dose intensity of chemotherapy actually received (RDI) and the prognosis. Preoperative chemotherapy consisted of high-dose methotrexate i.v., cisplatin i.a., and doxorubicin i.v. After surgery "good responder" patients (90% or more tumor necrosis) had a 31-weeks of chemotherapy with the same drugs, while "poor responder" patients (less than 90% tumor necrosis) received a 40 weeks treatment with ifosfamide and etoposide added to the three drugs used preoperatively. Due to delays and dose-reductions, only 17 patients (12%) received the treatment exactly as scheduled by the protocol, 66 (46%) received a dose-intensity between 90 and 99%, and 61 (42%) a dose-intensity between 63 and 89%. At a follow-up ranging between 10 and 13 years, 97 patients (67%) remained continuously free of disease, 45 relapsed, and two died of doxorubicin-induced cardiopathy. The continuous disease-free survival (CDFS) was not related to patients' gender and age, tumor histology, site and size, serum value of alkaline phosphatase, type of surgery and histologic response to chemotherapy. According to the RDI, CDFS resulted significantly higher for those 81 patients who received 90% or more of the scheduled dose-intensity than for those 61 who had less than 90% of the scheduled dose-intensity (76.5% v.s. 57.3%; p<0.02). These results seem to suggest that in neoadjuvant treatment of osteosarcoma the dose-intensity of chemotherapy is crucial for outcome, therefore every effort should be made to avoid reductions of doses and/or delays in performing the cycles of chemotherapy. PMID- 11410804 TI - K-ras gene mutations in non-small cell lung cancer in Japanese. AB - To evaluate the association of mutations in the K-ras gene with the incidence of non-small cell lung cancer (NSCLC) in Japanese patients, 410 patients treated surgically were studied. DNA was extracted from frozen specimens, and polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) hybridization assay was used to examine K-ras mutations in codons 12, 13 and 61. K-ras mutations were found in 33 patients (8.0%), and all were smokers or ex-smokers. There were no significant differences in clinical or pathological stage between wild-type cases and mutant cases. The most frequently identified mutation was a G ->T transversion (25/33, 75.8%) that resulted in the substitution of a glycine for a cysteine or a valine. Previous studies have shown that the majority of K ras mutations among smokers are G-->T transversion. In our study, K-ras mutations were found only in smokers, and there was a high frequency of G-->T transversions. A clear correlation exists between smoking and K-ras gene mutations. PMID- 11410805 TI - Combined effect of dietary calcium and iron on colonic aberrant crypt foci, cell proliferation and apoptosis, and fecal bile acids in 1,2-dimethylhydrazine treated rats. AB - To investigate the combined effect of Ca and Fe on colon carcinogenesis, cell proliferation, apoptosis and fecal bile acids, male Wistar rats were fed the diet containing 5 g Ca/kg (normal Ca) or 15 g Ca/kg (excessive Ca) with 45 mg Fe/kg (normal Fe) or 500 mg Fe/kg (excessive Fe) for 32 days, and given an injection of 1,2-dimethylhydrazine on day 4. Supplemental Ca reduced colonic aberrant crypt foci (ACF), especially in excess Fe group. Excessive Fe elevated the ACF, especially in the normal Ca diet. When the Ca intake was high, excessive Fe caused no influence on the ACF. Alteration of colonic ACF was associated with those of liver and serum Fe concentration. Also, colonic cell proliferation and concentration of deoxycholic acid (DCA) in fecal water-soluble fraction were reduced by supplementation of dietary Ca, but unaffected by that of dietary Fe. Supplementation of Ca and/or Fe elevated colonic cell apoptosis. The results suggest that dietary Ca markedly suppresses colon ACF in the Fe-overloaded rats through altering Fe status, and that supplemental Ca lowers colonic cell proliferation and fecal DCA in the water-soluble fraction and elevates colonic cell apoptosis irrespective of Fe status. PMID- 11410806 TI - Regulation of the growth of mouse Leydig cells by the inactive stereoisomer, 17alpha-estradiol: Lack of correlation between the elevated expression of ERalpha and difference in sensitivity to estradiol isomers. AB - We examined the effects of 17alpha-estradiol, the biologically inactive stereoisomer of 17beta-estradiol, on cell growth and cell cycle kinetics using the normalized mouse TM3 Leydig cells. A significant biphasic stimulatory growth response was observed by 17alpha-estradiol exposure with peaks at 1 pg/ml (157.13%) and 100 ng/ml (120.04%) (p<0.05). The growth stimulatory effects of 17alpha-estradiol were inhibited by tamoxifen. A significant decrease in cell cycle time of Leydig cells exposed to 17alpha-estradiol was observed in treated cells (p<0.05). RT-PCR analysis indicated that exposure to Leydig cells to 1 pg/ml and 100 ng/ml 17alpha-estradiol resulted in a 10- and 5-fold increases in the expression of ERalpha, respectively. Similar effects were observed with exposure to equivalent concentrations of 17beta-estradiol. Difference in sensitivity to stereoisomers of estradiol to growth response of Leydig cells did not correlate with the elevated expression of ERalpha. We conclude that the TM3 Leydig cells are sensitive to the non-typical estrogen, 17alpha-estradiol, presumably through the activation of ER-independent signaling transduction pathways. PMID- 11410807 TI - Expression of heparin binding epidermal growth factor-like growth factor in hepatocellular carcinoma: an immunohistochemical study. AB - Previous studies have demonstrated an elevated level of heparin-binding epidermal growth factor-like growth factor (HB-EGF) mRNA in hepatocarcinogenesis in a rodent model and of its protein as well as mRNA in human hepatocellular carcinoma (HCC). In order to elucidate the clinical significance of HB-EGF in human HCC, we immunohistochemically investigated its expression in 100 HCC cases with different characteristics. HB-EGF was not or only faintly expressed in non-cancerous hepatocytes. On the other hand, 59% of the HCC cases were classified as positive for HB-EGF, and its expression was more frequently observed in cases with low proliferating activity, at an early stage, of small size, without portal invasion, low alpha-fetoprotein level, without satellite nodules (borderline significance), and well or moderate differentiation. Furthermore, HB-EGF expression showed prognostic significance in both univariate and multivariate analyses and can be regarded as an independent prognostic factor of disease-free survival. These results strongly suggest the involvement of HB-EGF in early stage of HCC. PMID- 11410808 TI - Histological changes in the cervical lymph nodes after radiotherapy. AB - Radiotherapy to the neck is frequently employed in the treatment of malignancies, sometimes alone and sometimes before or after surgery or with or without chemotherapy. We checked the histopathological changes in the neck after radiotherapy, dividing the changes into two groups. One with short-term changes between six months to one year, and the second group with long-term changes after more than six years. We compared these two groups with a control group without radiotherapy. The changes in the long-term group were more pronounced than in the short-term group, with more stromal fibrosis, vascular changes, and specially lymph nodes smaller than one centimeter. Thus, bigger lymph nodes (more then one centimeter) are more radiosensitive and become smaller after six years. PMID- 11410809 TI - Cutaneous pseudolymphoma of the breast with late homozonal relapse. AB - Cutaneous pseudolymphomas refer to a group of lymphoproliferative disorders of difficult classification and diagnosis. Considered as very low-grade lymphomas by some authors, they are actually able to progress to overt cutaneous lymphomas in a minority of cases. We report a case of cutaneous pseudolymphoma of the breast in a 21-year old woman which relapsed locally ten years later. The case is of interest for the unusual localisation of the lesion, for its recurrence after such a long period of time, and for the homozonal site of recurrence. The reported features support both the long course and the lymphomatous nature of such lymphocytic proliferations. PMID- 11410810 TI - Microsatellite alterations in patients with thoracic sarcoma. AB - Few studies on sarcomas have examined the relationships between microsatellite alterations in particular loci, tumor prognosis and tumorigenesis, because sarcomas are uncommon and those prognoses can be confounded by coexisting factors, such as tumor site. We studied the relationship between microsatellite alterations and prognosis in 31 patients with thoracic sarcoma. The frequency of loss of heterozygosity (LOH) at 17p13 in stage IV sarcomas was significantly higher than that in stage I and III sarcomas (p<0.05). The 5-year survival for patients with LOH at 17p13 was significantly lower than that for patients without LOH (p<0.05). Six of 31 cases (19.4%) revealed replication error. These results suggest that p53 abnormality occurs during advanced stages of sarcoma and are related to patient prognosis, and it is possible that aberrations in mismatch repair activity are related to sarcoma tumorigenesis. PMID- 11410811 TI - Molecular analysis of p53 tumor-suppressor gene and microsatellites in preneoplastic and neoplastic lesions of the colon and esophagus. AB - Mutations in exons 4-8 of the p53 gene by the PCR-SSCP analysis in preneoplastic and neoplastic lesions of the colon (n=11) and esophagus (n=18) were screened. p53 overexpression by immunohistochemistry in 11 colonic lesions and 13 microsatellites, in all the patients (n=29), were also studied. A positive result concordancy between the three techniques was found in 1 adenoma and 2 adenocarcinomas of the colon, each with loss of heterozygocity of microsatellites. Metaplastic lesions of esophagus showed biallelic mutations and low frequency of microsatellite alterations. The relationship between genetic alterations in p53, microsatellites and type of colon and esophageal lesions is discussed. PMID- 11410812 TI - Surgical treatment of parathyroid carcinoma (Review). AB - Cancer of the parathyroid gland is rare and most patients suffer from hyperparathyroidism (HPT). Therefore it is often difficult to distinguish between benign HPT and malignant disorders. Prognosis as well as therapeutic approaches are still limited. Surgical treatment should be considered the therapy of choice for primary and recurrent manifestation of parathyroid carcinoma. The initial operation includes en-bloc resection of the tumor and the ipsilateral thyroid lobe. Surgery may cure some patients, but is often only palliative and aims to control hypercalcemia. In some patients hypercalcemia may be refractory to repeated surgery and medical treatment can provide short- or middle-term effects. PMID- 11410813 TI - Family lifestyle and parental body mass index as predictors of body mass index in Australian children: a longitudinal study. AB - OBJECTIVE: To investigate associations between body mass index (BMI) and family characteristics, including lifestyle, in parents and offspring from Australian families. DESIGN AND SUBJECTS: Longitudinal survey of 219 families of Australian children who had been surveyed 3-yearly between the ages of 9 and 18 y. MEASUREMENTS: Socio-economic status, weight and height, diet from 3 day records or food frequency questionnaires, alcohol consumption, smoking habits and physical fitness in offspring (bicycle ergometry in 18-y-olds). RESULTS: In 18-y olds, in models examining offspring's lifestyle variables, BMI was predicted negatively by physical fitness (P=0.012), and positively by alcohol intake (P=0.046) in sons while, in daughters, only a negative association with physical fitness was significant. In models including parental characteristics, BMI in 18 y-old sons and daughters was significantly predicted by mothers' and fathers' BMI, independently of offsprings' alcohol intake, smoking, physical fitness and parents' education, and, in daughters, by fathers' alcohol intake. These models explained 48% of variance in daughters and 33% in sons. In both sons and daughters, BMI over the 9 y of the survey was consistently higher in offspring with overweight or obese fathers (P=0.033 for sons, P=0.024 for daughters) or mothers (P=0.031 for sons, P=0.037 for daughters). Physical fitness at the ages of 12, 15 and 18 y was negatively related to fathers' obesity in daughters and mothers' obesity in sons. Obesity in fathers was associated with a four-fold increase in risk of obesity at the age of 18 y in both sons and daughters with an independent eight-fold increase in risk for daughters if mothers were obese. Birthweight was unrelated to overweight or obesity in the 18-y-olds. Alcohol intake in sons related significantly to alcohol intake in either parent while, for daughters, there was a significant association only with fathers' alcohol consumption. In daughters, fat intake was positively associated with fat intake score in both fathers and mothers. CONCLUSION: Parental overweight or obesity may identify children at risk for a range of unhealthy behaviours. Promotion of a healthy lifestyle targeting overweight families, particularly in lower socio economic groups, should be a priority. PMID- 11410814 TI - Overweight at age 21: the association with body mass index in childhood and adolescence and parents' body mass index. A cohort study of New Zealanders born in 1972-1973. AB - BACKGROUND: Obesity is an increasing problem so understanding the association between childhood and adolescent measures of body mass index (BMI) and being overweight at age 21 has implications for treatment or strategies to reduce its prevalence. OBJECTIVE: To examine the association between measures of BMI in childhood and adolescence and parents' BMI and being overweight at age 21. DESIGN: The study was based on a birth cohort born in Dunedin, New Zealand in 1972-1973. RESULTS: BMI tracked from childhood to early adulthood. The point on the BMI distribution where the probability of being overweight at age 21 was 0.5 was close to the 75th centile for boys throughout childhood and adolescence. It was rather higher for girls in childhood but similar in adolescence. Boys with a BMI above the 75th centile at age 7 were more than 4.0 times more likely to be overweight at age 21 than those with a BMI below the median. The relative risk for girls was 3.2. By age 15 this increased to 9.8 for males and 6.8 for females. Having overweight parents, particularly a mother, increased the likelihood of being overweight. Only 40% of those who were overweight at age 21 could be identified by age 7 and 25% were not identified until they were at least 15. CONCLUSION: Although a high BMI in childhood predicts being overweight at age 21, many of those who were overweight at age 21 had a BMI below the 75th centile or even the median in childhood and early adolescence. Population strategies, complemented by an individual approach for those above the 75th centile, are needed to reduce the average BMI of the population. PMID- 11410815 TI - Obesity in Auckland school children: a comparison of the body mass index and percentage body fat as the diagnostic criterion. AB - OBJECTIVES: First, to determine obesity rates in Auckland school children according to their ethnic group using two different criteria: the body mass index (BMI) and percentage body fat (PBF) derived from bioelectrical impedance analysis (BIA). Second to examine the relationship between BMI and body composition across ethnic groups to determine if BMI references from European children accurately reflect obesity in other ethnic groups. DESIGN: A total of 2273 Auckland school children, aged 5-10.9 y had their height, weight and bioelectrical impedance measured. Using these measurements, each child's BMI, fat free mass, fat mass and PBF were derived. RESULTS: In all 14.3% of children were obese using the recommended definition of obesity (BMI) greater than the 95th percentile). There was no clinically significant difference in the relationship between BMI and body composition in different ethnic groups. Obesity rates varied with ethnicity (P<0.0001) and were higher in Pacific Island (24.1%) and Maori (15.8%) than in European children (8.6%). Obesity rates also varied with age (P<0.03), with the highest rates in older children. PBF levels were higher in females than males (P<0.0001). Using a definition of obesity based on percentage body fat (PBF>30%), obesity rates were higher in all ethnic groups. CONCLUSIONS: Obesity rates are high in Auckland school children and there are clear differences in obesity rates in different ethnic groups. If BMI criteria are used to define obesity in our population, we recommend the same standards be used for children of all ethnicities. PMID- 11410816 TI - Prevalence and trends of obesity among school children in Taiwan--the Taipei Children Heart Study. AB - BACKGROUND: In Taiwan, the availability of a high-fat and high-energy diet has steadily risen over the past 30 y, while people have become increasingly sedentary. This lifestyle of poor diet and little physical activity has caused the prevalence of obesity to increase among adults and children. Obesity and associated chronic disease risk factors are becoming important public health issues. The purpose of this study is to evaluate the prevalence and secular trends of being overweight and obese among school children in Taiwan. METHODS: We conducted a cross-sectional survey among junior high school students in Taipei in 1994 to ascertain a representative distribution of demographic, lifestyle and biochemical characteristics, including several cardiovascular disease risk factors. After multistage sampling of 85 junior high schools in Taipei, we randomly selected 1500 children for this survey. We defined being overweight and obese based on ideal body weight (IBW) criteria. Children are considered to be overweight if their body weight is 110-120% of IBW, and obese if their body weight is greater than 120% of IBW within age- and gender-specific strata. RESULTS: In general, obese children have higher blood pressure, plasma glucose and triglyceride levels and lower high-density-lipoprotein cholesterol levels than normal-weight children. From 1980 to 1994, the mean value of body weight increased significantly over increases in body height (especially among boys). Although the percentage of overweight children remained steady from 1980 to 1994 in both genders, the prevalence and trends of obesity increased significantly, especially among boys and older girls. CONCLUSION: From this survey we find that in Taiwan from 1980 to 1994 body weight increased dramatically over body height among school children. The prevalence of obesity also increased significantly, especially among boys, while the percentage of overweight children did not vary. Overall, this study indicates that obesity and the adverse effects of being over the ideal body weight is no longer just a problem of Western countries. PMID- 11410817 TI - Body mass index of 0 to 45-y-old Danes: reference values and comparison with published European reference values. AB - OBJECTIVES: To generate body mass index (weight/height(2) (kg/m(2)), BMI) reference values for 0 to 45-y-old Danes and compare these with published European reference values. SUBJECTS: A national sample used to generate the current Danish height and weight reference (29 106 measurements made 1965-1977; age 0-21 y; sample I), and four samples from Copenhagen (3391 measurements made 1981-1985; age 7-45 y; samples II-III and 2608 measurements made 1991-1994; age 6 45 y; samples IV-V). DATA ANALYSIS: Using the LMS method, Danish BMI reference values by age and sex were constructed from samples I and II. These were compared with BMI reference values from Sweden (age 6-16 (girls) or 6-19 y (boys)), Germany (6-19 y), UK (0-23 y), and France (0-87 y). Two recently examined but smaller Danish cohorts (samples IV and V) were compared with the reference values to assess the secular trend in BMI. RESULTS: Overall, Danish BMI reference values (samples I and II) fitted best with French reference values and were systematically below UK, Swedish and German reference values. However, the BMI centiles of young adult Danish women were above French reference values and the BMI of Danes was substantially below French and UK reference values during the first months of life. The mean BMI Z-score of the recently examined samples was 0.24 (sample IV, P=0.0001) and 0.15 (sample V, P=0.0001) based on the French reference values and 0.19 (sample IV, P=0.0007) and 0.01 (sample V, P=0.49) based on the Danish reference values. CONCLUSION: For clinical purposes, we recommend comparing the BMI of Danish children and adolescents with the present Danish reference values and the new IOTF cut-off values and comparing the BMI of Danish adults with the WHO cut-off values. For research purposes, cut-off values and national or internationally used BMI reference values may be used, depending on the research questions. PMID- 11410818 TI - Prevalence of overweight and obesity in a rural southern Italy population and relationships with total and cardiovascular mortality: the Ventimiglia di Sicilia project. AB - OBJECTIVE: We investigated the prevalence of overweight and obesity and their relationships with the main cardiovascular risk factors in the population of Ventimiglia di Sicilia, a rural village in Southern Italy characterized by low cholesterol levels and by a low incidence of early coronary heart disease mortality. We related all deaths to body weight and fat distribution during an 8 y follow-up. DESIGN: Cross-sectional and prospective observational study. SUBJECTS: A total of 835 free-living individuals, 363 males and 472 females, of age between 20 and 69 y. MEASUREMENTS: In all participants body weight, waist-to hip ratio (WHR), cardiovascular risk factors and plasma lipids were measured. During the follow-up, total and cardiovascular deaths were registered. RESULTS: We found a high overall prevalence of subjects with overweight or obesity (respectively 45.0% and 27.7%), with great differences among classes of age. As expected, body weight and fat distribution were associated with diabetes, hypertension, dyslipidemia and with a worsening of lipid profile. During the follow-up we registered 37 total and 11 cardiovascular deaths. All-cause and cardiovascular mortality risks were, respectively, 1.64 (95% CI 0.65-4.15) and 2.71 (95% CI 0.29-25.26) in subjects with a body mass index (BMI) of 27-29.99 kg/m2 and 2.45 (95% CI 1.03-5.87) and 5.36 (95% CI 1.41-62.01) in subjects with a BMI of > or =30 kg/m2 in comparison with participants with a BMI of <27 kg/m2, and 3.48 (95% CI 1.46-8.30) and 4.55 (95% CI 1.12-18.40) in subjects with a WHR higher than the median in comparison with individuals with a WHR lower than the median. CONCLUSION: The Ventimiglia di Sicilia Study highlights the great importance of overweight and obesity as a public health issue in a rural population and indicates that it is necessary to consider the impact of body weight and fat distribution on both total and CHD mortality. PMID- 11410819 TI - Obesity, body fat distribution and coronary atherosclerosis among Japanese men and women. AB - OBJECTIVE: To investigate the relation of the obesity and body-fat distribution with angiographically defined coronary atherosclerosis. DESIGN: Cross-sectional study in a clinical setting. SUBJECTS: Three hundred and twenty men (median age, 59 y) and 212 women (median age, 67 y) who underwent coronary angiography for suspected or known coronary heart disease at 5 cardiology departments between September 1996 and August 1997. Patients with disease duration >1 y were excluded. MEASUREMENTS: The body mass index (BMI) and the waist to hip circumference ratio (WHR) were used as main exposure variables, and either the presence of significant coronary stenosis or the Gensini's score (> or =10 vs<10) as an outcome variable, in a sex-specific multiple logistic regression analysis controlling for age, hospital, and other coronary risk factors. RESULTS: Among male patients, BMI was progressively higher with an increasing number of vessels involved (P trend=0.05); the adjusted odds ratios for the presence of significant stenosis across quartiles of BMI were 1.0 (reference), 1.1, 1.9 and 2.5 (P trend=0.02), and the positive association was more pronounced for younger patients. Among females, however, such associations were not evident. Employing the Gensini's score as an outcome gave similar results. WHR was not significantly associated with either outcome regardless of sex. CONCLUSION: These results suggested that BMI was predictive of coronary stenosis among male patients, but not among female patients. Unlike most previous studies, this study failed to detect a positive association with WHR. PMID- 11410820 TI - Body fat, fat distribution and serum lipids, lipoproteins and apolipoproteins in African-American and Caucasian-American prepubertal children. AB - OBJECTIVE: The purpose of the present study was to determine the impact of body fat mass and fat distribution on serum lipids, lipoproteins and apolipoproteins in African-American and Caucasian-American prepubertal children. SUBJECTS: Study participants included 62 African-American children (age 8.3+/-1.4 y; body mass 37.3+/-13.6 kg; height 133+/-11 cm) and 39 Caucasian children (age 8.6+/-1.2 y; body mass 34.1+/-11.0 kg; height 131+/-9 cm). METHODS: Venous blood samples were obtained after a 12 h overnight fast and serum was analyzed for total cholesterol (TC), high-density lipoprotein-cholesterol (HDL-C), triacylglycerol (TAG), apolipoprotein A-I (ApoA-I), apolipoprotein B (ApoB) and lipoprotein (a) (Lp(a)) concentrations. Body composition and body fat distribution were measured by dual energy X-ray absorptiometry and computed tomography, respectively. RESULTS: African-American children had lower TAG (46+/-20 vs 61+/-32 mg/dl, P=0.015) and higher Lp(a) (34+/-25 vs 17+/-28 mg/dl, P=0.001) and HDL-C (44+/-11 vs 39+/-8 mg/dl, P=0.041). There were no ethnic differences in TC, ApoA-I and ApoB (P=0.535, P=0.218, P=0.418, respectively). The ethnic difference in TAG and Lp(a) was not explained by total fat or abdominal fat. The ethnic difference in HDL-C was explained by visceral fat and TAG. CONCLUSION: In prepubertal children, neither body fat nor fat distribution explain the ethnic difference in TAG or Lp(a), but visceral fat and TAG may contribute to differences in HDL-C. PMID- 11410821 TI - Reduced oxidized low-density lipoprotein after weight reduction in obese premenopausal women. AB - BACKGROUND: Several studies support the hypothesis that oxidation of low-density lipoprotein (LDL) promotes atherogenesis. Obesity is one of the risk factors of atherosclerosis, but it is not known whether obesity is related to LDL oxidation. OBJECTIVE AND DESIGN: We investigated the effect of weight reduction and subsequent weight maintenance program on LDL oxidation in 77 obese premenopausal women (BMI 29-46 kg/m(2)). Another group of seven obese women served as a control group. Oxidized LDL was measured as baseline concentration of conjugated dienes in LDL lipids (ox-LDL). The weight reduction was performed in 12 weeks, using a very-low-energy diet. RESULTS: The mean weight loss was 13 kg (92 vs 79 kg). During weight reduction, the concentration of LDL cholesterol decreased by 11%, the concentration of ox-LDL decreased by 40%, and the ratio of ox-LDL to LDL by 33%. The concentration of LDL antioxidant capacity (LDL-TRAP) decreased by 8%, but the decrease was caused by the decrease in LDL. The concentration of LDL, ox LDL or LDL-TRAP did not change in the control group. The weight reduction correlated with the decrease of ox-LDL. During the subsequent 9 month weight maintenance programme, the concentrations of serum LDL (10%), ox-LDL (11%), LDL TRAP (29%), and the ratio of LDL-TRAP to LDL (21%) decreased. CONCLUSION: This study strengthens the evidence that the risk of atherogenesis is influenced favourably by weight reduction in obese women. This risk reduction is associated with a reduced oxidation of LDL. PMID- 11410822 TI - Weight change and blood coagulability and fibrinolysis in healthy obese women. AB - OBJECTIVES: To study the effect of weight loss and subsequent weight maintenance or weight regain on the activities of FVII and plasminogen activator inhibitor 1 (PAI-1) and the concentration of fibrinogen over 12 months in obese women consuming a hypoenergetic, low-fat diet with or without orlistat. In addition, the relation between the changes of the activities of PAI-1 and FVII with the changes of other cardiovascular risk factors were examined. METHODS AND PROCEDURES: Design-a 12-month randomized double-blind weight reduction trial of placebo and orlistat. Subjects-51 healthy obese women (age 44+/-0.7 y, BMI 36.2+/ 0.5 kg/m(2), mean+/-s.e.m.) Treatment-the participants were on a hypoenergetic diet (-600 kcal daily). The diet was adjusted for actual body weight (-300 kcal) at 6 months. Women were randomized to receive either orlistat 120 mg three times daily (n=25) or placebo three times daily (n=26) for 12 months according to a double-blind protocol after a 1 month run-in period. Measurements-changes of body weight, body composition, haemostatic and other cardiovascular risk factors were measured at 3-6 month intervals. The activity of plasma PAI-1 was measured by a chromogenic method, fibrinogen by the PT-derived method and the activity of FVII by the one-stage method. RESULTS: The changes in body weight between orlistat and placebo groups were not statistically significantly different. Orlistat did not influence haemostatic factors beyond its effect on weight loss. Therefore, the results of the orlistat and placebo groups were pooled. The average weight loss at 3, 6 and 12 months was 7.6, 9.5 and 10.0 kg, respectively (P<0.001). Between 6 and 12 months, 35% of women regained weight, 24% had stable weight and 41% continued to lose weight. No changes in the mean plasma fibrinogen concentration were observed at any time point during the trial. During the first 3 months the activities of PAI-1 and FVII decreased. The decline depended on the magnitude of weight loss. Between months 6 and 12 the changes of PAI-1 and FVII activities paralleled the changes of body weight. The activities rose with weight rebound but remained below the 6-month values if weight loss was sustained or continued. The changes of serum insulin were significantly correlated with the changes of both PAI-1 and FVII at 6 months and with PAI-1 at 12 months. CONCLUSIONS: The maintenance of modest weight loss is associated with long-term benefits in PAI-1 and FVII in obese women. The change of serum insulin is associated with the changes of PAI-1 activities. Fibrinogen is not affected by modest weight loss. PMID- 11410823 TI - Elevated homocysteine levels with weight loss after Lap-Band surgery: higher folate and vitamin B12 levels required to maintain homocysteine level. AB - OBJECTIVE: To investigate homocysteine levels and their relationship with serum folate and vitamin B12 concentrations with weight loss after the Lap-Band form of gastric restrictive surgery, with the view to minimizing risk. METHODS: We measured levels of fasting plasma homocysteine (tHcy), folate (serum and RBC) and vitamin B12 in two groups. The study group was 293 consecutive patients at 12 (n=192) or 24 (n=101) months review after surgery. The controls were 244 consecutive patients presenting for this surgery. RESULTS: The group losing weight had higher geometric mean tHcy levels: 10.4 (95% CI, 9.8-10.8) micromol/l compared with 9.2 (95% CI, 8.9-9.7) in controls (P<0.001). This occurred with higher folate levels and unchanged vitamin B12 levels. Levels of folate and B12 together explained 35% (r (2)) of the homocysteine variance in the weight loss group compared with only 9% (r (2)) in controls (P<0.001). Those taking regular multivitamin supplements had lower tHcy levels: 9.6 (9.1-10.0) micromol/l vs 12.3 (11.4-13.3) in those not taking supplements (P<0.001). A low normal plateau of tHcy levels occurred at levels of folate >15 ng/l and B12)600 ng/ml. A curvilinear relationship exists between these cofactors and tHcy levels, with the dose-response relationship shifted to the right in the weight loss group. CONCLUSION: This study shows elevated tHcy levels with weight loss, without lower serum folate or vitamin B(12) levels. There is an altered dose-response relationship with higher serum B(12) and folate levels required to maintain recommended tHcy levels. Patients losing weight have significant health benefits; however, they may be at greater risk of vascular events or fetal abnormality in association with raised tHcy levels. Multivitamin supplementation is effective in lowering tHcy levels. PMID- 11410824 TI - Visceral fat and coronary pathology in male adolescents. AB - OBJECTIVE: To find out whether coronary atherosclerotic lesions and their precursors in male adolescents are associated with the amount of mesenteric and omental fat. SUBJECTS: A series of 40 forensic autopsy cases of ante-mortem healthy boys of 13-19 y of age were investigated. METHODS: Body height and weight, waist and hip circumferences and the thickness of the abdominal subcutaneous fat were measured, the body mass index (BMI) and waist-to-hip ratio (WHR) were calculated, and omental and mesenteric fat deposits were weighed. The intimal surface of the coronary arteries covered by lesions was measured by planimetry, and the thickness of the intima was measured by computerized image analysis. Intimal macrophage foam cells and smooth muscle cells were detected by immunohistochemisty, and macrophages were quantified. RESULTS: The intima thickness of the left anterior descending artery (LAD) and in the thickest lesion varied significantly across the tertiles of visceral fat when adjusted for age, being highest when the sum weight of omental and mesenteric fat exceeded 358 g. The intima thickness of the circumflex artery (CX) varied significantly across the tertiles of waist circumference when adjusted for age. No statistically significant associations with other indicators of obesity were found. Macrophage foam cells were present in the lesions and their maximal density/mm(2) correlated significantly with intima thickness in the LAD and CX. The maximal density of macrophages in CX and the right coronary artery (RCA) and in the thickest lesion varied significantly across the tertiles of visceral fat when adjusted for age, being highest when the amount of fat exceeded 358 g. The macrophage density also varied significantly across the tertiles of waist circumference in all vessels. CONCLUSIONS: The results indicate that early macrophage-rich coronary lesions are associated with increased amounts of visceral fat in adolescent male individuals. This emphasizes the importance of effective prevention of weight gain in individuals with a tendency to accumulate visceral fat at an early age. PMID- 11410825 TI - Left ventricular mass and function in young obese women. AB - OBJECTIVE: Obesity is associated with a high mortality rate due to cardiovascular disease. Left ventricular (LV) hypertrophy has been described in relation to obesity. The aim of this study was to evaluate echocardiographically the LV mass and function in young obese women as compared to lean women with similar characteristics. DESIGN: Prospective study. SUBJECTS: Eighty-two young women (< or =40 y), with obesity degree varying from I to III (BMI from 30 to 50 kg/m2) were compared to eighty young lean women. All of them were normotensive, none had cardiovascular complaints or any previous history of pulmonary disease, and none were taking any medication. The LV mass was calculated by the Devereux and Reichek formula. RESULTS: The LV mass was strongly increased in all obese groups (P<0.00003 to 0.000005) compared to lean subjects. LV mass adjusted indexes for height, BMI or volume were also increased compared to lean subjects and when adjusted for weight it was decreased. However when comparing LV mass/body surface area index this difference was not statistically significant. The linear regression analysis showed a strong association between the degree of obesity and LV mass, (r=0.52, P<0.001). Systolic and diastolic function in obese patients were similar to lean subjects, except for a lower E/A ratio in the obese group (P=0.005). CONCLUSION: In asymptomatic young obese women, there are some echocardiographic findings suggesting early cardiac involvement that seems to be related to the degree of obesity. PMID- 11410826 TI - Insulin, renin-aldosterone system and blood pressure in obese people. AB - OBJECTIVE: To evaluate the relationship between insulin, the renin-aldosterone system and blood pressure in obese subjects. DESIGN AND METHODS: A cross sectional study of a group of severely obese normotensive subjects who were surgical candidates (n=39; mean BMI: 47.8+/-1.4) and a group of hypertensive patients (n=57; mean BMI: 28.0+/-0.7) twenty-nine of whom had BMI>27. All subjects were studied after 15 days on a balanced diet. Insulin, plasma renin activity and aldosterone were measured. RESULTS: Fasting insulin, plasma renin activity and aldosterone were higher in severely obese normotensive subjects than in hypertensive subjects (respectively 32.3+/-3.0 vs 13.1+/-1.0 mU/l, P=0.0001; 1.34+/-0.22 vs 0.88+/-0.12 ng/ml/h, P=0.04; 137.2+/-16.2 vs 87.9+/-12.1 pg/ml, P=0.015). Insulin was related to BMI and to aldosterone both in normotensive and in hypertensive patients. CONCLUSION: Hyperinsulinemia itself does not determine hypertension; in some people it could play a vasodilator role in opposition to the renin-aldosterone system. PMID- 11410827 TI - Endocrine, metabolic and nutritional factors in obesity and their relative significance as studied by factor analysis. AB - OBJECTIVE: To evaluate different aspects of obesity, such as body fat distribution, plasma hormone and lipid profiles, adipose tissue composition and dietary intake in an obese population in order to identify the most important factors that contribute to obesity. DESIGN: Eighty-five obese subjects, 30 men and 55 women (age, 30-70 y; body mass index (BMI), 27-35 kg/m2), were studied using anthropometric measurements, computed tomography, adipose tissue composition, serum hormone and lipid profiles and nutritional evaluations. To determine to what extent individual factors contributed to the general process of obesity, the data were subjected to a factor analysis. RESULTS: Three patterns of anthropometric and computed tomography data emerged that accounted for 69% of the variance. Factor 1 defined abdominal obesity and explained 30% of the total variance, factor 2 (gynoid obesity) accounted for 26%; and factor 3 (subcutaneous fat) explained 13% of the total variance. When other factors associated with obesity, such as lipid profile, hormonal profile and fat composition, were introduced, obesity itself, especially abdominal obesity, remained the principal factor, accounting for 23% of total variability. All factors were of secondary importance when dietary characteristics were introduced. In the overall factor analysis, more than 40% of the variability in obesity was related to dietary habits, particularly fat intake, followed by energy and saturated fatty acids intake. CONCLUSION: Even though obesity is a multifactorial phenomenon, the results suggest that dietary intake, especially fat intake, is the most important factor contributing to obesity. Secondary factors include endocrine and metabolic factors. PMID- 11410828 TI - Fat phobia scale revisited: the short form. AB - OBJECTIVE: To develop a shortened form of the original 50-item fat phobia scale. METHOD: The first factor from the original fat phobia scale-undisciplined, inactive and unappealing-was identified as a potential short form of the scale. A new sample of 255 people completed the original 50-item scale. The reliability of a shortened 14-item version of the scale was tested and compared to that of the full scale using both the new sample and the original sample of 1135 study participants. RESULTS: The fat phobia scale-short form demonstrated excellent reliability in both samples and was strongly correlated with the 50-item scale. Mean and 90th percentile scores are given for both the long and short versions of the scale. CONCLUSION: The shortened fat phobia scale is expected to increase the utility of the measure in a diverse array of research and clinical settings. Future research should focus on developing scale norms for the general population and conducting research on fat phobia in males and among different ethnic groups. PMID- 11410829 TI - Metabolic correlates of eating behavior in severe obesity. AB - BACKGROUND: The benefit of spreading energy intake over many small meals ('nibbling') rather than few large ones ('gorging') for control of blood glucose, serum lipids and body fat accretion has been known for 60 y, but the mechanisms are poorly understood. Men exhibit more of a gorging eating pattern than women and are also more prone to the metabolic complications of obesity, as are women with a 'male', central distribution of adipose tissue. We have shown correlations between central fat distribution, and other components of the metabolic 'Syndrome X' and fatty infiltration of the liver. Here we study relationships between eating rate and fat distribution and test the hypothesis that gorging might be associated with fatty liver. SUBJECTS AND METHODS: In 30 non-alcoholic, non diabetic, severely obese women (body mass index, BMI=47+/-1 kg/m(2); mean+/ s.e.m.) with a mean age of 36+/-1 y and 16 men (BMI: 52+/-3) age 38+/-2 y, who were candidates for anti-obesity surgery, we measured eating rate using an eating monitor, and fat distribution by the waist-hip circumference ratio (WHR). In addition in the 17 women and 11 men who had surgery, serum lipids were analyzed and routine liver biopsies were evaluated for steatosis by a pathologist blinded to the conditions of the study. RESULTS: Men ate significantly faster than women (188+/-28 vs 123+/-9 g/min; P<0.01), and had more liver fat (score: 2.7+/-03 vs 1.5+/-0.3; P<0.01), with no statistically significant sex differences in s cholesterol or s-triglycerides. Eating rate correlated with WHR (r=0.46; P<0.01, n=46), liver fat (r=0.55; P<0.01), and s-triglycerides (r=0.42; P<0.05) adjusting for sex. Liver fat correlated with WHR (r=0.50; P<0.05), s-triglycerides (r=0.70; P<0.01) and s-cholesterol (r=0.50; P<0.05), while there were no significant correlations with BMI or body weight. In multivariate analysis eating rate (32%), meal size (8%) and WHR (6%) contributed 46% of the variance in liver fat. CONCLUSION: We showed increased eating rates in severely obese men and women with central fat distribution. Furthermore, increased eating rates were associated with fatty liver and elevated serum lipids. Eating rate in severely obese women and men may be a determinant of the metabolic syndrome. PMID- 11410830 TI - Abnormalities of bioimpedance measures in overweight and obese hemodialyzed patients. AB - BACKGROUND: The body composition in overweight and obese hemodialyzed patients (HD) remains ill-defined. This study evaluates in HD patients the influence of body size, as indicated by body mass index (BMI, kg/m(2)), on body composition by measuring bioimpedance analysis (BIA)-derived variables (phase angle (PA), fat free mass (FFM) and body cell mass (BCM). METHODS: We studied 50 Caucasian patients (mean age 62.8+/-9.2 y) on standard bicarbonate hemodialysis for at least 12 months who regularly achieved dry weight in post-HD, received similar dialysis doses and were free from inflammation/infection. Thirty-eight gender- and age-matched healthy subjects were included as controls (CON). Both HD and CON were divided into three groups on the basis of their BMI(kg/m2) 18.5-24.9, normal weight (NW); 25-29.9, overweight (OW); and > or =30, obese (OB). In HD patients, BIA was performed 30 min after the end of dialysis. RESULTS: Seven patients were obese (12%) while 16 were overweight (32%); in CON, 12 were obese (31%) and 12 overweight (31%). BIA-measured extracellular water was comparable in all groups. PA, which was similar in normal-weight HD and CON (6.2+/-0.9 degrees and 6.3+/ 0.8 degrees ), decreased in OW- and OB-HD patients (5.3+/-1.0 degrees and 5.2+/ 0.6 degrees, respectively; P<0.05 vs NW-HD) while it was unchanged in OW- and OB CON (6.1+/-0.8 degrees and 5.9+/-0.5 degrees, P<0.05 vs respective HD groups). In OW and OB patients, the lower PA values were coupled with a major reduction of BIA-derived percentage BCM and FFM (P<0.05 vs NW-HD, and vs OW- and OB-CON). In patients, PA and BCM correlated with anthropometry-measured FFM. Of note, serum albumin and protein catabolic rate were significantly reduced in OB patients. CONCLUSION: In overweight and obese HD patients, BIA-derived FFM, BCM and PA are significantly lower with respect to normal-weight patients and BMI-matched controls. These abnormalities of body composition are coupled with reduction of anthropometric measures of lean mass and a decrease of protein intake that, however, becomes significant only in the obese. We therefore suggest that overweight and obese HD patients are at risk of protein malnutrition in spite of excessive energy intake. BIA may be considered as a useful diagnostic tool to detect such a condition early. PMID- 11410831 TI - Foot-to-foot bioelectrical impedance analysis: a valuable tool for the measurement of body composition in children. AB - OBJECTIVE: To determine the accuracy of foot-to-foot bioelectrical impedance analysis (BIA) and anthropometric indices as measures of body composition in children. DESIGN: Comparison of foot-to-foot BIA and anthropometry to dual-energy X-ray absorptiometry (DEXA)-derived body composition in a multi-ethnic group of children. SUBJECTS: : Eighty-two European, NZ Maori and Pacific Island children aged 4.9-10.9 y. MEASUREMENTS: DEXA body composition, foot-to-foot bioelectrical impedance, height, weight, hip and waist measurements. RESULTS: Using a BIA prediction equation derived from our study population we found a high correlation between DEXA and BIA in the estimation of fat-free mass (FFM), fat mass (FM) and percentage body fat (PBF) (r=0.98, 0.98 and 0.94, respectively). BIA-FFM underestimated DEXA-FFM by a mean of 0.75 kg, BIA-FM overestimated DEXA-FM by a mean of 1.02 kg and BIA-PBF overestimated DEXA-PBF by a mean of 2.53%. The correlation between six anthropometric indices (body mass index (BMI), ponderal index, Chinn's weight-for-height index, BMI standard deviation score, weight-for length index and Cole's weight-for-height index) and DEXA were also examined. The correlation of these indices with PBF was remarkably similar (r=0.85-0.87), more variable with FM (r=0.77-0.94) and poor with FFM (r=0.41-0.75). CONCLUSIONS: BIA correlated better than anthropometric indices in the estimation of FFM, FM and PBF. Foot-to-foot BIA is an accurate technique in the measurement of body composition. PMID- 11410832 TI - Does the BMI reflect body fat in obese children and adolescents? A study using the TOBEC method. AB - OBJECTIVE: Due to the fact that obesity is defined as excess of body fat mass, we tested the hypothesis whether the body mass index (BMI) can be used as a valid measure for the detection of the degree of obesity in individual obese children and adolescents. METHODS: A total of 204 obese children and adolescents (105 boys, 99 girls) aged 6-17 y, using total body electrical conductivity (TOBEC) for fat measurement, were included into this study. A multiple regression analysis was performed with percentage body fat (PBF) as dependent variable and BMI, age and sex as independent variables. First- and second-order interaction terms were also included. Since all interaction terms showed a significant influence on PBF, regression analysis was performed separately for boys and girls, dividing each group into two age subgroups (subjects younger than 10 y, and subjects 10 y or older). RESULTS: BMI and PBF were observed to be positively correlated (overall: r=0,65; P=0.0001; boys r=0.63 and girls: r=0.68). Through a multiple regression analysis 57% of the variance of PBF could be explained by the independent variables. In boys younger than 10 y 73% and in girls younger than 10 y 63% of the variance of PBF was explained by the BMI. In subjects 10 y or older the association was poor (boys: 27%; girls: 38%). It should be emphasized that there is a wide range in the relationship between PBF and BMI in the obese subjects. CONCLUSION: From these results we conclude that BMI might be a useful parameter for epidemiological studies: however in the individual pediatric patient, especially from 10 y onwards, it gives only a limited insight to the degree of obesity based on the definition. PMID- 11410833 TI - The prediction of body composition in Chinese Australian females. AB - OBJECTIVE: To determine the efficacy of applying specific body composition techniques to, and assess the relationship between body mass index (BMI) and body fat levels for Chinese Australian females. DESIGN: Statistical comparative analysis of body composition techniques. SUBJECTS: Australian resident females of Chinese extraction (n=40) (aged 18-45 y, mean 32.5+/-8.0; BMI range 15.7-30.9 kg/m2, mean 21.7+/-3.1 kg/m2, median 20.8 kg/m2). MEASUREMENTS: Body composition determined using bio-electrical impedance analysis (BIA), the skin-fold equations of Durnin and Womersley (D&W) and a deuterium dilution technique. Body size was calculated as the body mass index (BMI) weight/height(2) (kg/m2). RESULTS: With a median BMI of 20.8, range 15.7-30.9, an acceptable BMI existed for 87.5% of the subjects (mean (s.d.) 21.7+/-3.1 kg/m2). Percentage fat mass (%FM) from the deuterium dilution technique (mean (s.d.) 35.6+/-6.4) suggested 75% were overweight or obese. %FM from the D&W equation (mean (s.d.) 28.0+/-3.9) and BIA (mean (s.d.) 29.4+/-5.1) also indicated a tendency towards overweight or obese. The deuterium technique was significantly correlated and significantly different to the D&W eqn, r=0.71 P=0.001; and BIA, r=0.77, P=0.001. Bland and Altman analysis indicated that bias existed between the techniques (BIA mean (s.d.) 6.7+/-4.1) and D&W equation mean (s.d.)-6+/-4.5) when compared to the deuterium method. CONCLUSIONS: Despite a low mean BMI, body fat levels determined by the three methods suggested that, overall, an unsatisfactory body composition existed. The levels of overweight and obesity (%FM>30) were higher than reported in previous research despite a mean BMI lower than the Australian national average. Comparative analysis suggested that the body fat prediction techniques used may be precise but not accurate. Comparative results obtained for the BIA and D&W equation techniques suggest an overestimation of body fat levels for leaner individuals and under estimation for overfat individuals. The results support the notion that accurate determination of body composition and the determination of appropriate body size may require equations developed for specific ethnic populations. PMID- 11410834 TI - Enhanced intra-abdominal visceral fat accumulation in patients with Werner's syndrome. AB - OBJECTIVE: Studies were made on the abnormality of glucose and lipid metabolism and its cause in four patients with Werner's syndrome to infer the reason for accelerated atherogenesis in this syndrome. RESULTS: Of these four patients, hypercholesterolemia was found in three, hypertriglyceridemia in four, hypoalphalipoproteinemia in two and hypertension in two. All the patients had insulin-resistant diabetes mellitus and three of them had apparent hyperinsulinemia. Abdominal computed tomography revealed that all of them had visceral fat obesity, namely augumented intra-abdominal adipose tissue. CONCLUSION: The clinical features of these patients resemble those recently designated as insulin resistant syndrome (syndrome X) or visceral fat syndrome. The metabolic abnormality may be one of important factors in the accelerated atherogenesis in this syndrome. PMID- 11410835 TI - Relationships between tamoxifen use, liver fat and body fat distribution in women with breast cancer. AB - Tamoxifen is a nonsteroidal anti-estrogenic drug used for adjuvant treatment of breast cancer and recently as a chemopreventative agent for breast cancer and, on an investigational basis, for other cancers. To date there are case reports of hypertriglyceridemia and fatty liver disease in tamoxifen users. Fatty liver is associated with visceral obesity and other components of the metabolic syndrome. Here we evaluated steatosis and adipose tissue distribution by CT scan in a cross sectional study of 32 women on tamoxifen and 39 control women. Tamoxifen users had more visceral adipose tissue (VAT) and more liver fat than controls. This is the first study to demonstrate that fatty liver and intra-abdominal fat accumulation are common in breast cancer patients receiving tamoxifen. Prospective studies of tamoxifen should monitor metabolic changes in obese women with or without breast cancer. PMID- 11410836 TI - Obesity and borderline personality symptomatology: comparison of a psychiatric versus primary care sample. AB - This study explores the relationship between obesity and borderline personality symptomatology in two clinical settings: a psychiatric vs primary care setting. The body mass indices (BMI) of 48 women from a psychiatric outpatient setting and 83 women from a primary care setting were calculated. Each participant completed the borderline personality scale of the Personality Diagnostic Questionnaire Revised (PDQ-R). While BMI and PDQ-R were moderately related in the psychiatric sample (r=0.43, P<0.01), there was a lack of association between these variables in the primary care sample (r=0.04, P>0.05). In conclusion, women's increasing body weight appears to have some degree of correlation to borderline personality symptomatology among psychiatric patients, whereas it apparently does not among primary care patients. PMID- 11410837 TI - Linkage disequilibrium in humans: models and data. AB - In this review, we describe recent empirical and theoretical work on the extent of linkage disequilibrium (LD) in the human genome, comparing the predictions of simple population-genetic models to available data. Several studies report significant LD over distances longer than those predicted by standard models, whereas some data from short, intergenic regions show less LD than would be expected. The apparent discrepancies between theory and data present a challenge both to modelers and to human geneticists-to identify which important features are missing from our understanding of the biological processes that give rise to LD. Salient features may include demographic complications such as recent admixture, as well as genetic factors such as local variation in recombination rates, gene conversion, and the potential segregation of inversions. We also outline some implications that the emerging patterns of LD have for association mapping strategies. In particular, we discuss what marker densities might be necessary for genomewide association scans. PMID- 11410838 TI - Somatic mosaicism in hemophilia A: a fairly common event. AB - Mutations in the large gene of clotting factor VIII (FVIII) are the most common events leading to severe human bleeding disorder. The high proportion of de novo mutations observed in this gene raises the possibility that a significant proportion of such mutations does not derive from a single germ cell but instead should be attributed to a germline or somatic mosaic originating from a mutation during early embryogenesis. The present study explores this hypothesis by using allele-specific PCR to analyze 61 families that included members who had sporadic severe hemophilia A and known FVIII gene defects. The presence of somatic mosaicisms of varying degrees (0.2%-25%) could be shown in 8 (13%) of the 61 families and has been confirmed by a mutation-enrichment procedure. All mosaics were found in families with point mutations (8 [25%] of 32 families). In the subgroup of 8 families with CpG transitions, the percentage with mosaicism increased to 50% (4 of 8 families). In contrast, no mosaics were observed in 13 families with small deletions/insertions or in 16 families with intron 22 inversions. Our data suggest that mosaicism may represent a fairly common event in hemophilia A. As a consequence, risk assessment in genetic counseling should include consideration of the possibility of somatic mosaicism in families with apparently de novo mutations, especially families with the subtype of point mutations. PMID- 11410839 TI - Genomewide linkage analysis of stature in multiple populations reveals several regions with evidence of linkage to adult height. AB - Genomewide linkage analysis has been extremely successful at identification of the genetic variation underlying single-gene disorders. However, linkage analysis has been less successful for common human diseases and other complex traits in which multiple genetic and environmental factors interact to influence disease risk. We hypothesized that a highly heritable complex trait, in which the contribution of environmental factors was relatively limited, might be more amenable to linkage analysis. We therefore chose to study stature (adult height), for which heritability is approximately 75%-90% (Phillips and Matheny 1990; Carmichael and McGue 1995; Preece 1996; Silventoinen et al. 2000). We reanalyzed genomewide scans from four populations for which genotype and height data were available, using a variance-components method implemented in GENEHUNTER 2.0 (Pratt et al. 2000). The populations consisted of 408 individuals in 58 families from the Botnia region of Finland, 753 individuals in 183 families from other parts of Finland, 746 individuals in 179 families from Southern Sweden, and 420 individuals in 63 families from the Saguenay-Lac-St.-Jean region of Quebec. Four regions showed evidence of linkage to stature: 6q24-25, multipoint LOD score 3.85 at marker D6S1007 in Botnia (genomewide P<.06), 7q31.3-36 (LOD 3.40 at marker D7S2195 in Sweden, P<.02), 12p11.2-q14 (LOD 3.35 at markers D12S10990-D12S398 in Finland, P<.05) and 13q32-33 (LOD 3.56 at markers D13S779-D13S797 in Finland, P<.05). In a companion article (Perola et al. 2001 [in this issue]), strong supporting evidence is obtained for linkage to the region on chromosome 7. These studies suggest that highly heritable complex traits such as stature may be genetically tractable and provide insight into the genetic architecture of complex traits. PMID- 11410840 TI - Quantitative-trait-locus analysis of body-mass index and of stature, by combined analysis of genome scans of five Finnish study groups. AB - In recent years, many genomewide screens have been performed, to identify novel loci predisposing to various complex diseases. Often, only a portion of the collected clinical data from the study subjects is used in the actual analysis of the trait, and much of the phenotypic data is ignored. With proper consent, these data could subsequently be used in studies of common quantitative traits influencing human biology, and such a reanalysis method would be further justified by the nonbiased ascertainment of study individuals. To make our point, we report here a quantitative-trait-locus (QTL) analysis of body-mass index (BMI) and stature (i.e., height), with genotypic data from genome scans of five Finnish study groups. The combined study group was composed of 614 individuals from 247 families. Five study groups were originally ascertained in genetic studies on hypertension, obesity, osteoarthritis, migraine, and familial combined hyperlipidemia. Most of the families are from the Finnish Twin Cohort, which represents a population-wide sample. In each of the five genome scans, approximately 350 evenly spaced markers were genotyped on 22 autosomes. In analyzing the genotype data by a variance-component method, we found, on chromosome 7pter (maximum multipoint LOD score of 2.91), evidence for QTLs affecting stature, and a second locus, with suggestive evidence for linkage to stature, was detected on chromosome 9q (maximum multipoint LOD score of 2.61). Encouragingly, the locus on chromosome 7 is supported by the data reported by Hirschhorn et al. (in this issue), who used a similar method. We found no evidence for QTLs affecting BMI. PMID- 11410841 TI - High-resolution multipoint linkage-disequilibrium mapping in the context of a human genome sequence. AB - A new method is presented for fine-scale linkage disequilibrium (LD) mapping of a disease mutation; it uses multiple linked single-nucleotide polymorphisms, restriction-fragment-length polymorphisms, or microsatellite markers and incorporates information from an annotated human genome sequence (HGS) and from a human mutation database. The method takes account of population demographic effects, using Markov chain Monte Carlo methods to integrate over the unknown gene genealogy and gene coalescence times. Information about the relative frequency of disease mutations in exons, introns, and other regions, from mutational databases, as well as assumptions about the completeness of the gene annotation, are used with an annotated HGS, to generate a prior probability that a mutation lies at any particular position in a specified region of the genome. This information is updated with information about mutation location, from LD at a set of linked markers in the region, to generate the posterior probability density of the mutation location. The performance of the method is evaluated by simulation and by analysis of a data set for diastrophic dysplasia (DTD) in Finland. The DTD disease gene has been positionally cloned, so the actual location of the mutation is known and can be compared with the position predicted by our method. For the DTD data, the addition of information from an HGS results in disease-gene localization at a resolution that is much higher than that which would be possible by LD mapping alone. In this case, the gene would be found by sequencing a region < or =7 kb in size. PMID- 11410842 TI - Variant 1859G-->A (Arg620Gln) of the "hairless" gene: absence of association with papular atrichia or androgenic alopecia. PMID- 11410843 TI - The presence of mitochondrial haplogroup x in Altaians from South Siberia. PMID- 11410851 TI - Dynamics of pyruvate metabolism in Lactococcus lactis. AB - The pyruvate metabolism in the lactic acid bacterium Lactococcus lactis was studied in anaerobic cultures under transient conditions. During growth of L. lactis in continuous culture at high dilution rate, homolactic product formation was observed, i.e., lactate was produced as the major end product. At a lower dilution rate, the pyruvate metabolism shifted towards mixed acid-product formation where formate, acetate, and ethanol were produced in addition to lactate. The regulation of the shift in pyruvate metabolism was investigated by monitoring the dynamic behavior of L. lactis in continuous cultures subjected to step changes in dilution rate. Both shift-up and shift-down experiments were carried out, and these experiments showed that the enzyme pyruvate formate-lyase (PFL) plays a key role in the regulation of the shift. Pyruvate formate-lyase in vivo activity was regulated both at the level of gene expression and by allosteric modulation of the enzyme. A simple mathematical model was proposed to estimate the relative significance of the regulatory mechanisms involved. PMID- 11410852 TI - Biohydrogen generation by mesophilic anaerobic fermentation of microcrystalline cellulose. AB - Sixteen batch experiments were performed to evaluate the stability, kinetics, and metabolic paths of heat-shocked digester (HSD) sludge that transforms microcrystalline cellulose into hydrogen. Highly reproducible kinetic and metabolic data confirmed that HSD sludge could stably convert microcrystalline cellulose to hydrogen and volatile fatty acids (VFA) and induce metabolic shift to produce alcohols. We concluded that clostridia predominated the hydrogen producing bacteria in the HSD sludge. Throughout this study the hydrogen percentage in the headspace of the digesters was greater than 50% and no methanogenesis was observed. The results emphasize that hydrogen significantly inhibited the hydrogen-producing activity of sludge when initial microcrystalline cellulose concentrations exceeded 25.0 g/L. A further 25 batch experiments performed with full factorial design incorporating multivariate analysis suggested that the ability of the sludge to convert cellulose into hydrogen was influenced mainly by the ratio of initial cellulose concentration (So) to initial sludge density (Xo), but not by interaction between the variables. The hydrogen producing activity depended highly on interaction of So x (So/Xo). Through response surface analysis it was found that a maximum hydrogen yield of 3.2 mmol/g cellulose occurred at So = 40 g/L and So/Xo = 8 g cellulose/g VSS. A high specific rate of 18 mmol/(g VSS-d) occurred at So = 28 g/L and So/Xo = 9 g cellulose/g VSS. These experimental results suggest that high hydrogen generation from cellulose was accompanied by low So/Xo. PMID- 11410853 TI - Expression of GnTIII in a recombinant anti-CD20 CHO production cell line: Expression of antibodies with altered glycoforms leads to an increase in ADCC through higher affinity for FC gamma RIII. AB - The gene encoding the rat glycosylation enzyme beta1-4-N acetylglucosaminyltransferase III (GnTIII) was cloned and coexpressed in a recombinant production Chinese hamster ovary (CHO) cell line expressing a chimeric mouse/human anti-CD20 IgG1 antibody. The new cell lines expressed high levels of antibody and have growth kinetics similar to that of the parent. Relative QPCR showed the cell lines to express varying levels of mRNA. High performance liquid chromatography (HPLC) analysis showed the enzyme to have added bisecting N-acetylglucosamine (GlcNAc) residues in most (48% to 71%) of the N linked oligosaccharides isolated from antibody preparations purified from the cell lines. In an ADCC assay the new antibody preparations promoted killing of CD20-positive target cells at approximately 10- to 20-fold lower concentrations than the parent. This activity was blocked using an anti-Fc gamma RIII antibody, supporting the role of Fc gamma RIII binding in this increase. In addition, cell binding assays showed the modified antibody bound better to Fc gamma RIII expressing cells. The increase in ADCC activity is therefore likely due to an increased affinity of the modified antibody for the Fc gamma RIII receptor. PMID- 11410854 TI - Effect of crown ethers on structure, stability, activity, and enantioselectivity of subtilisin Carlsberg in organic solvents. AB - Colyophilization or codrying of subtilisin Carlsberg with the crown ethers 18 crown-6, 15-crown-5, and 12-crown-4 substantially improved enzyme activity in THF, acetonitrile, and 1,4-dioxane in the transesterification reactions of N acetyl-L-phenylalanine ethylester and 1-propanol and that of (+/-)-1 phenylethanol and vinylbutyrate. The acceleration of the initial rate, V(0), ranged from less than 10-fold to more than 100-fold. All crown ethers activated subtilisin substantially, which excludes a specific macrocyclic effect from being responsible. The secondary structure of subtilisin was studied by Fourier transform infrared (FTIR) spectroscopy. 18-Crown-6 and 15-crown-5 led to a more nativelike structure of subtilisin in the organic solvents employed when compared with that of the dehydrated enzyme obtained from buffer alone. However, the high level of activation with 12-crown-4 where this effect was not observed excluded overall structural preservation from being the primary cause of the observed enzyme activation. The conformational mobility of subtilisin was investigated by performing thermal denaturation experiments in 1,4-dioxane. Although only a small effect of temperature on subtilisin structure was observed for the samples prepared with or without 12-crown-4, both 18-crown-6 and 15-crown-5 caused the enzyme to denature at quite low temperatures (38 degrees C and 56 degrees C, respectively). No relationship between this property and V(0) was evident, but increased conformational mobility of the protein decreased its storage stability. The possibility of a "molecular imprinting" effect was also tested by removing 18 crown-6 from the subtilisin-18-crown-6 colyophilizate by washing. V(0) was only halved as a result of this procedure, an effect insignificant compared with the ca. 80-fold rate enhancement observed prior to washing in THF. This suggests that molecular imprinting is likely the primary cause of subtilisin activation by crown ethers, as recently suggested. PMID- 11410855 TI - Regeneration of adenosine triphosphate from glycolytic intermediates for cell free protein synthesis. AB - A new approach for adenosine triphosphate (ATP) regeneration in a cell-free protein synthesis system is described. We first show that pyruvate can be used as a secondary energy source to replace or supplement the conventional secondary energy source, phosphoenol pyruvate (PEP). We also report that glucose-6 phosphate, an earlier intermediate of the glycolytic pathway, can be used for ATP regeneration. These new methods provide more stable maintenance of ATP concentration during protein synthesis. Because pyruvate and glucose-6-phosphate are the first and last intermediates of the glycolytic pathway, respectively, the results also suggest the possibility of using any glycolytic intermediate, or even glucose, for ATP regeneration in a cell-free protein synthesis system. As a result, the methods described provide cell-free protein synthesis with greater flexibility and cost efficiency. PMID- 11410856 TI - Continuous production of (R)-phenylacetylcarbinol in an enzyme-membrane reactor using a potent mutant of pyruvate decarboxylase from Zymomonas mobilis. AB - The optimization of a continuous enzymatic reaction yielding (R) phenylacetylcarbinol (PAC), an intermediate of the L-ephedrine synthesis, is presented. We compare the suitability of three pyruvate decarboxylases (PDC), PDC from Saccharomyces cerevisiae, PDC from Zymomonas mobilis, and a potent mutant of the latter, PDCW392M, with respect to their application in the biotransformation using acetaldehyde and benzaldehyde as substrates. Among these, the mutant enzyme was the most active and most stable one. The reaction conditions of the carboligation reaction were investigated by determining initial rate velocities with varying substrate concentrations of both aldehydes. From the resulting data a kinetic model was inferred which fits the experimental data with sufficient reliability to deduce the optimal concentrations of both substrates for the enzymatic process. The results demonstrate that the carboligation is most efficiently performed using a continuous reaction system and feeding both aldehydes in equimolar concentration. Initial studies using a continuously operated enzyme-membrane reactor gave (R)-PAC with a space-time yield of 81 g L( 1). d(-1) using a substrate concentration of 50 mM of both aldehydes. The yield was easily increased by cascadation of enzyme-membrane reactors. The new strategy allows the synthesis of (R)-PAC from cheap substrates in an aqueous reaction system. It thereby overcomes the limitation of by-product formation that severely limits the current fermentative process. PMID- 11410857 TI - Performance of batch membrane reactor: Glycerol-3-phosphate synthesis coupled with adenosine triphosphate regeneration. AB - Glycerol-3-phosphate (G3P) was synthesized from glycerol using glycerol kinase (GK). This reaction requires adenosine triphosphate (ATP) and was coupled with the ATP regeneration reaction using acetate kinase (AK) in a batch-operated ultrafiltration hollow-fiber reactor. By taking into consideration the dynamic nature of the bioreactor performance under non-steady-state conditions, a model for the performance of a batch membrane reactor for G3P synthesis coupled with ATP regeneration was developed and studied. The simulation results showed good agreement with the experimental results. The simulation studies have provided some insight into the process dynamics of the coupled reactions in the reactor system studied. For the reactor operational model used, in which the enzymes are retained in the shell side and the substrates are also initially placed in the shell side, it was found that the substrate concentration in the lumen side increased to a level higher than that in the shell side, and a backdiffusion occurred from the lumen side to the shell side during reactor operation. The ratio of the reaction rate to diffusion rate goes through a sharp peak during the time that the direction of diffusion is reversed. For another reactor operational model, in which the substrates were initially placed in the lumen side and enzymes were retained in the shell side, it was found that the rate-controlling step between the reaction and diffusion was switched during the reactor operation. Initially, the reaction rate increased while the diffusion rate was high and the substrate concentrations increased in the shell side. The ratio of reaction rate to diffusion rate increased to a maximum and remained at a constant level as the diffusion rate decreased to a low level due to the nonlinear characteristics of mass transfer process. This study provides information that is useful for optimization of batch membrane enzyme reactor operation and for a fed batch-type process with an intermittent feeding strategy for efficient use of substrates. PMID- 11410858 TI - Linkers for improved cleavage of fusion proteins with an engineered alpha-lytic protease. AB - Addition of an N-terminal fusion partner can greatly aid the expression and purification of a recombinant protein in Escherichia coli. We investigated two genetically engineered proteases designed to remove the fusion partner after the protein of interest has been expressed. Recombinant human insulin-like growth factor-II (hIGF-II) has been produced from E. coli-derived fusion proteins using a novel enzymatic cleavage system that uses a mutant of alpha-lytic protease. Initially, two potential fusion protein linkers were designed, Pro-Ala-Pro-His (PAPH) and Pro-Ala-Pro-Met (PAPM), and were tested as substrates in the form of synthetic dodecapeptides. Using mass spectrometry and reverse-phase HPLC, the position of cleavage was confirmed and the kinetics of synthetic peptide cleavage were examined. Use of the linkers in hIGF-II fusion proteins produced in E. coli was then evaluated. The fusion proteins constructed consist of the first 11 amino acids of porcine growth hormone linked N-terminally to hIGF-II by six amino acids that include the dipeptide Val-Asn followed by a variable tetrapeptide protease cleavage motif. Mass spectrometry and N-terminal sequencing confirmed that proteolytic cleavage of the fusion proteins had occurred at the predicted sites. Using the fusion proteins as substrates, the cleavage of the rationally designed motifs by the alpha-lytic protease mutant was compared. The fusion protein containing the motif PAPM had a k(cat)/K(M) ratio indicating a 1.6-fold preference over the PAPH fusion protein for cleavage by this enzyme. Furthermore, when hIGF-II fusion proteins containing the designed cleavable linkers were processed with the engineered alpha-lytic protease, they gave greatly improved yields of native hIGF-II compared to an analogous fusion protein cleaved by H64A subtilisin. Comparison of the peptide and protein cleavage studies shows that the efficient proteolysis of the cleavage motifs is an inherent property of the designed sequences and is not determined by secondary or tertiary structure in the fusion proteins. PMID- 11410859 TI - Analysis of single-chain antibody production in Pichia pastoris using on-line methanol control in fed-batch and mixed-feed fermentations. AB - In the last few years the Pichia pastoris expression system has been gaining more and more interest for the expression of recombinant proteins. Many groups have employed fermentation technology in their investigations because the system is fairly easy to scale up and suitable for the production in the milligram to gram range. A large number of heterologous proteins from different sources has been expressed, but the fermentation process technology has been investigated to a lesser extent. A large number of fermentations are carried out in standard bioreactors that may be insufficiently equipped to meet the demands of high-cell density fermentations of methylotrophic yeasts. In particular, the lack of on line methanol analysis leads to fermentation protocols that may impair the optimal expression of the desired products. We have used a commercially available methanol sensor to investigate in detail the effects of supplementary glycerol feeding while maintaining a constant methanol concentration during the induction of a Mut(+) strain of Pichia pastoris. Specific glycerol feed rates in the range of 38-4.2 mg. g(-1). h(-1) (mg glycerol per gram fresh weight per hour) were investigated. Expression of the recombinant scFv antibody fragment was only observed at specific feed rates below 6 mg. g(-1). h(-1). At low specific feed rates, growth was even lower than with methanol as the sole carbon source and the harvest expression level of the scFv was only half of that found in the control fermentation. These results show that glycerol inhibits expression driven by the AOX1 promoter even at extremely limited availability and demonstrate the benefits of on-line methanol control in Pichia fermentation research. PMID- 11410860 TI - Second primary neoplasms in 633,964 cancer patients in Sweden, 1958-1996. AB - The Swedish Family-Cancer Database was used to analyze concordant (same site) and discordant (different site) second primary neoplasms in 633,964 cancer patients diagnosed from 1958 to 1996. Cases of second malignant neoplasms were extracted from the Database if the diagnosis date of the first and second cancer differed by at least 1 month. The expected numbers of cancers were obtained by applying site-, sex-, age-, period-, residence- and socioeconomic level-specific rates in the corresponding population in the Database to the appropriate person-years at risk. The standardized incidence ratio (SIRs) of a second cancer was taken to be the ratio of observed to expected numbers of second cancers. Of all cancers, 8.5% were subsequent neoplasms (8.4% for males and 8.7% for females). SIRs for both concordant and discordant subsequent cancer were elevated in patients with cancer of the upper aerodigestive tract, colon, nose, breast, other female genitals, testis, kidney, urinary, bladder, skin, nervous system, endocrine, bone, connective tissue, melanoma, lymphoma and leukemia. The risks at some concordant sites, such as nose, squamous cell skin, bone and connective tissue in both sexes, breast in males and upper aerodigestive tract and leukemia in females, were very high (>10). At discordant sites, SIRs were less than 2 but significantly increased after all but gastric and prostatic cancer. Compared with the general population, cancer patients were at a modestly increased risk for new primary cancer after cancers at many sites, calling for attention in treatment, management and prevention. PMID- 11410861 TI - Quantitative real-time RT-PCR detection of breast cancer micrometastasis using a multigene marker panel. AB - Real-time RT-PCR is a relatively new technology that uses an online fluorescence detection system to determine gene expression levels. It has the potential to significantly improve detection of breast cancer metastasis by virtue of its exquisite sensitivity, high throughput capacity and quantitative readout system. To assess the utility of this technology in breast cancer staging, we determined the relative expression levels of 12 cancer-associated genes (mam, PIP, mamB, CEA, CK19, VEGF, erbB2, muc1, c-myc, p97, vim and Ki67) in 51 negative-control normal lymph nodes and in 17 histopathology-positive ALNs. We then performed a receiver operating characteristic (ROC) curve analysis to determine the sensitivity and specificity levels of each gene. Areas under the ROC curve indicated that the most accurate diagnostic markers were mam (99.6%), PIP (93.3%), CK19 (91.0%), mamB (87.9%), muc1 (81.5%) and CEA (79.4.0%). mam was overexpressed in 16 of 17 lymph nodes known to contain metastatic breast cancer at levels ranging from 22- to 2.8 x 10(5)-fold above normal mean expression, whereas PIP was overexpressed from 30- to 2.2 x 10(6)-fold above normal in 13 lymph nodes. Real-time RT-PCR analysis of pathology-negative LN from breast cancer patients revealed evidence of overexpression of PIP (6 nodes), mam (3 nodes) and CEA (1 node) in 8 of 21 nodes (38%). Our results provide evidence that mam, PIP, CK19, mamB, muc1 and CEA can be applied as a panel for detection of metastatic and occult micrometastatic disease. PMID- 11410862 TI - Antisense oligonucleotides targeting the epidermal growth factor receptor inhibit proliferation, induce apoptosis, and cooperate with cytotoxic drugs in human cancer cell lines. AB - We have constructed a series of 22 phosphorothioate 20-mer antisense oligonucleotides directed against different regions of the human (EGFR) mRNA. Treatment with EGFR antisense oligonucleotides showed a dose-dependent inhibition of human GEO colon cancer cell growth in soft agar. Western blot analysis demonstrated a significant reduction in EGFR expression after treatment with each EGFR antisense oligonucleotide. The ability to inhibit GEO anchorage-independent growth, however, varied among the EGFR antisense sequences with an IC(50) ranging between 0.5 and 3.5 microM. Two of these antisense oligonucleotides targeting the regions between 2457-2476 and 614-4633 bases of the human EGFR mRNA have been modified as hybrid DNA/RNA mixed backbone oligonucleotides (MBO) to examine their anticancer properties in vivo. The 2 EGFR antisense MBOs retained the same biological properties of the fully phosphorothioate EGFR antisense oligonucleotides targeting the same EGFR mRNA sequences, such as blocking EGFR synthesis, inhibiting cell growth and enhancing programmed cell death in human cancer cell lines that express functional EGFRs. Furthermore, a potentiation in the growth inhibitory effect on GEO cancer cells was observed after treatment with these EGFR antisense MBOs in combination with cytotoxic drugs, including cisplatin, doxorubicin, paclitaxel, or topotecan. These results show the antiproliferative activity of specific EGFR antisense oligonucleotides and allow to identify novel EGFR antisense MBOs that deserve further evaluation as potential selective anticancer agents alone or in combination with cytotoxic drugs in human carcinomas that express functional EGFRs. PMID- 11410863 TI - Expression of protein kinase C beta1 confers resistance to TNFalpha- and paclitaxel-induced apoptosis in HT-29 colon carcinoma cells. AB - The expression of different protein kinase C (PKC) isoenzymes has been shown to vary with proliferation rates, differentiation or apoptosis in normal colon crypts. In addition, the activity of some PKC isoenzymes appears to be reduced in colorectal cancer. The aim of the present work was to determine whether modulation of PKC expression would affect the susceptibility of a p53-defective colon carcinoma cell line to different apoptotic treatments. HT-29 cells exhibited sensitivity to paclitaxel (Taxol) and tumor necrosis factor alpha (TNFalpha) in a dose- and time-dependent manner but were relatively resistant to etoposide. Inhibition of PKC activity augmented the susceptibility of HT-29 cells to apoptosis, and phorbol ester induction of PKC reduced such susceptibility. Transfected HT-29(PKC) cells, hyper-expressing the beta1 isoform of PKC, were less sensitive to TNFalpha and paclitaxel than the normal counterpart. The present data 1) indicate that the expression of PKC influences the susceptibility of HT-29 colon cancer cells to apoptotic drugs apparently regardless of their mechanism of action, and 2) suggest paclitaxel as a potential candidate for the treatment of colon cancer, possibly in association with inhibitors of PKC (alpha and beta) at doses not cytotoxic per se. PMID- 11410864 TI - CD95-induced JNK activation signals are transmitted by the death-inducing signaling complex (DISC), but not by Daxx. AB - Here we investigated CD95-mediated JNK activation pathways and their physiological relevance by employing a variety of cell lines with deficiencies in individual signal transmitting proteins. JNK activation was completely dependent on the activation of caspases in type I and type II cells, as revealed by the inhibitory effects of the caspase inhibitors zVAD-fmk or the cowpoxvirus-encoded CrmA protein. Jurkat cells deficient in caspase-8 or expressing a dominant negative (DN) form of FADD were unable to induce JNK in response to CD95 ligation, indicating that these death-inducing signaling complex (DISC) proteins are required for signal transmission. Activation of caspases, JNK and apoptosis occurred with a markedly slower kinetics in cells expressing a DN version of ASK1, revealing an important contribution of ASK1 for these processes. A C terminally truncated version of Daxx impaired CD95-mediated apoptosis without affecting the JNK signal. DN forms of FADD, MKK4 and MKK7 completely inhibited CD95-mediated JNK activation but remained without impact on cell killing, indicating that JNK activation is not required for the execution process of CD95 mediated cell killing. PMID- 11410865 TI - Expression of caspases-3, -6 and -8 and their relation to apoptosis in non-small cell lung carcinoma. AB - We analyzed a set of 103 non-small cell lung carcinomas (NSCLCs) for caspase-3, 6 and -8 expression and apoptosis. Additionally, the expression of bcl-2, bax and p53 were studied. Caspase-3 positivity appeared as diffuse, cytoplasmic staining and was restricted to the tumor area. In contrast, the immunoreactivity for caspase-6 was intense, granular and mostly located in single cells or groups of tumor cells showing apoptotic morphology. The caspase-8 expression pattern was a combination of the two other caspases studied, featuring both diffuse and single cell patterns restricted to the tumor area. No significant differences were seen in caspase -3, -6 and -8 expression between tumors of different histological types or grades. The number of apoptotic cells and bodies was significantly higher in NSCLCs, in which caspase-8 immunostaining was mainly seen in single cells (p = 0.017), whereas caspase -3 and -6 expression had no association with apoptosis. It is apparent that, in lung tissue, up-regulation of caspase expression is a phenomenon associated solely with neoplasia and reflects the readiness of the tumor cells to undergo apoptosis. Interestingly, caspases -3, -6 and -8 each have an individual staining pattern in NSCLC, perhaps reflecting their different position in the caspase hierarchy. PMID- 11410866 TI - Role of p53 in the responses of human urothelial cells to genotoxic damage. AB - Loss of p53 function is a feature of many types of malignancy, including transitional-cell carcinoma (TCC), where it is associated with high-grade lesions and the development of muscle-invasive disease. Genotoxic agents used as part of the treatment strategy may contribute to tumour progression by inducing further non-lethal DNA damage in surviving cells. To determine the role of p53 in cellular responses to genotoxic agents, we used cultured normal human urothelial (NHU) cells and NHU cells with disabled p53 function. Mitomycin C and gamma radiation caused normal cells to undergo an extended period of cell-cycle arrest, followed by complete recovery of proliferative potential. In contrast, cells with disabled p53 function, whether karyotypically normal (HU-E6 cells) or post-crisis with karyotypic abnormalities (HU-E6P cells), underwent extensive apoptosis. Overall survival was dose-dependent, and surviving HU-E6 cells from low-dose treatments showed clonal karyotypic abnormalities. These findings demonstrate that p53 status is a crucial factor in determining the ability of urothelial cells to survive DNA damage and suggest caution in the use of genotoxic treatments for low-grade tumours as our data imply that malignancies that have not yet lost p53 function will show the same "repair-and-recovery" response as normal cells. PMID- 11410867 TI - TGF-beta-induced invasiveness of pancreatic cancer cells is mediated by matrix metalloproteinase-2 and the urokinase plasminogen activator system. AB - TGF-beta strongly promotes local tumor progression in advanced epithelial tumors, though the underlying mechanisms are poorly understood. In the present study, we demonstrate the potential of TGF-beta to increase the invasiveness of the pancreatic cancer cell lines PANC-1 and IMIM-PC1. TGF-beta-induced tumor cell invasion occurred in a time-dependent manner, started after 12 hr and continued to increase even 48 hr after a single application of the growth factor. Blocking of secreted TGF-beta1 by application of neutralizing antibodies 24 hr after TGF beta treatment completely prevented the sustained effects of TGF-beta on tumor cell invasion. Together with our previous observation that TGF-beta1 up-regulates its own expression in both cell lines, our data suggest that TGF-beta1 acts in an autocrine manner to maintain tumor cell invasion. As measured by Northern blot hybridization and zymography, TGF-beta treatment of PANC-1 and IMIM-PC1 cells resulted in strong up-regulation of expression and activity of both matrix metalloproteinase-2 (MMP-2) and the urokinase plasminogen activator (uPA) system. Treatment with MMP inhibitors or inhibitors of the uPA system caused significant reduction of TGF-beta-induced invasiveness in both cell lines. In contrast, expression and activity of MMP-2 and uPA as well as tumor cell invasiveness remained unaffected in cell lines with defects of the TGF-beta type II receptor (MiaPaca2) or the Smad4 gene (IMIM-PC2 and CAPAN-1). In these cell lines, TGF beta also failed to auto-induce its own expression. In conclusion, our results suggest that TGF-beta1 is a strong promotor of pancreatic cancer progression. TGF beta thereby acts in an autocrine manner to induce tumor cell invasion, which is mediated by MMP-2 and the uPA system. PMID- 11410868 TI - Real-time observation of micrometastasis formation in the living mouse liver using a green fluorescent protein gene-tagged rat tongue carcinoma cell line. AB - Initial arrest, attachment, extravasation and subsequent extravascular growth of tumor cells in the secondary organs are believed to be crucial events for hematogenous metastasis, but the actual processes in living animals remain unclear. For the present study, we established green fluorescent protein (GFP) expressing rat tongue carcinoma cell lines (RSC3) that permit real-time analysis of micrometastasis formation in combination with intravital video microscopy (IVVM). With this system, GFP-expressing metastatic (LM-EGFP) and non-metastatic (E2-EGFP) cell lines could be visualized at the cellular level in live mice for more than 1 month. Real-time IVVM analysis of liver metastases after intraportal injection of cells via a mesenteric vein revealed that both LM-EGFP and E2-EGFP tumor cells arrest similarly in sinusoidal vessels near terminal portal venules within 0.4 sec, during which time no evidence of a "rolling"-like movement along endothelial cell surfaces was observed. Quantitative analysis of GFP-positive foci showed that E2-EGFP cells were completely sheared from the liver sinusoid within 3 days, with no solitary dormant cells, whereas a substantial number of LM EGFP cells remained in the liver, probably due to stable attachment to the sinusoidal wall. Confocal laser scanning microscopic study in combination with laminin immunohistochemistry revealed that only LM-EGFP cells started growth at 3 to 4 days after inoculation and that most of the growing foci were surrounded by subsinusoidal basement membrane. Our results suggest that micrometastasis formation by LM-EGFP cells consists of initial tumor cell arrest due to size constraints of the vessel, stable attachment to subsinusoidal basement membrane and subsequent intravascular growth before extravasation. The difference in metastatic potential between the 2 lines may reside in their capacity to attach stably to the vessel wall rather than their potential for initial cell arrest or subsequent growth. The system used in the present study may be a powerful tool for analyzing targets for various anti-metastatic agents in the sequential process of metastasis. PMID- 11410869 TI - Induction of apoptosis by cyclo-oxygenase-2 inhibitor NS398 through a cytochrome C-dependent pathway in esophageal cancer cells. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) can induce tumor cells to undergo apoptosis in vitro. They have also shown cancer-preventive activity in vivo. The mechanism of their effects is, however, not well defined. We investigated the mechanism by which a new NSAID, NS398, induces apoptosis in esophageal cancer cell lines. NS398 decreased cell viability in 2 cyclo-oxygenase-2-positive (COX 2(+)) esophageal cancer cell lines but not in a COX-2(-) cell line. DNA fragmentation and TUNEL assays demonstrated that NS398 induced the 2 COX-2(+) cancer cell lines to undergo apoptosis. The percentage of apoptosis induced by NS398 was associated with the level of COX-2 expression. Further investigation showed that the cytochrome c pathway was responsible for NS398-induced apoptosis; i.e., cytochrome c was released from mitochondria, caspase-9 and caspase-3 were activated and finally poly(ADP-ribose)polymerase (PARP) was cleaved. Furthermore, the effect of NS398 was inhibited by the caspase inhibitor Z-DEVD-FMK and prostaglandin E(2). In contrast, bcl-2, bax, c-myc, Fas and Fas-ligand showed minor changes. Altogether, our data suggest that induction of apoptosis by NS398 is associated with COX-2 expression and occurs through the cytochrome c-dependent pathway, which sequentially activates caspase-9 and caspase-3 and cleaves PARP. PMID- 11410870 TI - Sensitization to TNF-induced apoptosis by 1,25-dihydroxy vitamin D(3) involves up regulation of the TNF receptor 1 and cathepsin B. AB - The active form of vitamin D(3), 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), induces caspase-independent apoptosis in MCF-7 and T47D breast cancer cells. Before the appearance of apoptotic cells at Day 4 after the addition of 1,25(OH)(2)D(3), the MCF-7 cells are sensitized to the caspase-mediated apoptosis induced by TNF. We studied the mechanism underlying the cross talk between these 2 distinct death pathways in MCF-7 and T47D cells. Whereas 1,25(OH)(2)D(3) pre treatment enhanced TNF-induced apoptosis of TNF sensitive MCF-7 cells, it failed to render TNF resistant T47D cells sensitive to this cytokine. Opposing to an earlier report suggesting that cytosolic phospholipase A(2) (cPLA(2)) mediates the 1,25(OH)(2)D(3)-induced sensitization to TNF, we could not detect any cPLA(2) protein in MCF-7 cells and its overexpression had no effect on cellular sensitivity to 1,25(OH)(2)D(3) or the combination with TNF. The sensitization of MCF-7 cells to TNF-induced apoptosis by pre-treatment with 1,25(OH)(2)D(3) may instead be partially explained by an increased surface expression of the TNF receptor 1 (TNF-R1). In line with this, not only the TNF-induced activation of caspases and apoptosis but also that of NF-kappaB was enhanced by 1,25(OH)(2)D(3) pre-treatment. Furthermore, 1,25(OH)(2)D(3) enhanced TNF-induced NF-kappaB activation in T47D cells suggesting that it potentiates TNF signaling in general. Interestingly, the lysosomal protease cathepsin B, which expression is up regulated by 1,25(OH)(2)D(3), was released from the lysosomes upon TNF treatment, and inhibition of its activity rescued 1,25(OH)(2)D(3) treated MCF-7 cells from TNF-induced apoptosis. In conclusion, 1,25(OH)(2)D(3) may enhance TNF-induced apoptosis by increasing the expression of both the TNF-R1 and cathepsin B. PMID- 11410871 TI - Biological evidence that human papillomaviruses are etiologically involved in a subgroup of head and neck squamous cell carcinomas. AB - High-risk human papillomaviruses (HPVs) have been proposed to be associated with a subset of head and neck cancers (HNSCCs). However, clear biological evidence linking HPV-mediated oncogenesis to the development of HNSCC is hardly available. An important biological mechanism underlying HPV-mediated carcinogenesis is the inactivation of p53 by the HPV E6 oncoprotein. In the present study we investigated this biological relationship between HPV and HNSCC. In total 84 HNSCC tumors were analyzed for the presence of high-risk HPV nucleic acids by DNA polymerase chain reaction-enzyme immunoassay (PCR-EIA) and E6 reverse transcriptase (RT)-PCR as well as for the presence of mutations in the p53 gene. We found 20/84 HPV16 DNA-positive cases with one or more DNA assays, 10 of which were consistently positive with all assays. Only 9/20 cases showed E6 mRNA expression, indicative for viral activity. Only these nine E6 mRNA-positive cases all lacked a p53 mutation, whereas both the other HPV DNA-positive and HPV-DNA negative tumors showed p53 mutations in 36% and 63% of the cases, respectively. Moreover, only in lymph node metastases of HPV E6 mRNA-positive tumors both viral DNA and E6 mRNA were present. Our study provides strong biological evidence for a plausible etiological role of high-risk HPV in a subgroup of HNSCC. Analysis of E6 mRNA expression by RT-PCR or alternatively, semiquantitative analyses of the viral load, seem more reliable assays to assess HPV involvement in HNSCC than the very sensitive DNA PCR analyses used routinely. PMID- 11410872 TI - Kinetics of cytokine expression in melanoma metastases classifies immune responsiveness. AB - Production of cytokines (CKs) in the tumor micro-environment may modulate tumor host interactions. However, pre-clinical models often provide conflicting data and there is no established role for CKs as modulators of the natural or treatment-related behavior of tumors. Serial sampling by fine-needle aspirates (FNAs) of identical metastases from patients affected with metastatic melanoma and undergoing IL-2-based vaccination allowed prospective measurement of IL-10, TGF-beta1, TGF-beta2 and IFN-gamma transcriptional levels assessed by quantitative real-time PCR. Thus, it was possible to prospectively document the expression of markers relevant to a given treatment and follow at the same time the clinical outcome of the lesions left in place. Eight of 27 metastatic lesions completely regressed in response to the treatment and 1 demonstrated >50% shrinkage. These regressions occurred after the follow-up FNA had been obtained. IL-10 transcript was differentially expressed in pre-treatment FNA of responding lesions (t-test p(2) = 0.002). During treatment, INF-gamma transcript levels significantly increased in regressing compared to non-regressing lesions (t-test p(2) = 0.03). These data suggest that the pre-treatment CK profile of the tumor micro-environment may determine clinical responsiveness to immune therapy. Furthermore, temporal changes in CK expression during treatment might describe the biological characteristics of an effective immune response. PMID- 11410873 TI - A comparison of two types of dendritic cell as adjuvants for the induction of melanoma-specific T-cell responses in humans following intranodal injection. AB - Dendritic cells (DCs) elicit potent anti-tumoral T-cell responses in vitro and in vivo. However, different types of DC have yet to be compared for their capacity to induce anti-tumor responses in vivo at different developmental stages. Herein, we correlated the efficiencies of different types of monocyte-derived DC as vaccines on the resulting anti-tumor immune responses in vivo. Immature and mature DCs were separately pulsed with a peptide derived from tyrosinase, MelanA/MART-1 or MAGE-1 and a recall antigen. Both DC populations were injected every 2 weeks in different lymph nodes of the same patient. Immune responses were monitored before, during and after vaccination. Mature DCs induced increased recall antigen-specific CD4(+) T-cell responses in 7/8 patients, while immature DCs did so in only 3/8. Expansion of peptide-specific IFN-gamma-producing CD8(+) T cells was observed in 5/7 patients vaccinated with mature DCs but in only 1/7 using immature DCs. However, these functional data did not correlate with the tetramer staining. Herein, immature DCs also showed expansion of peptide-specific T cells. In 2/4 patients vaccinated with mature DCs, we observed induction of peptide-specific cytotoxic T cells, as monitored by chromium-release assays, whereas immature DCs failed to induce peptide-specific cytotoxic T cells in the same patients. Instead, FCS-cultured immature DCs induced FCS-specific IgE responses in 1 patient. Our data demonstrate that this novel vaccination protocol is an efficient approach to compare different immunization strategies within the same patient. Thus, our data define FCS-free cultured mature DCs as superior inducers of T-cell responses in melanoma patients. PMID- 11410874 TI - Over-expression of APAF-1 and caspase-9 augments radiation-induced apoptosis in U 373MG glioma cells. AB - The p53 tumor-suppressor gene plays a critical role in radiation-induced apoptosis. Several genes, including Bax and Fas, are involved in p53-mediated apoptosis, and their over-expression enhances the degree of radiation-induced apoptosis. Apaf-1 and caspase-9 have been reported to be downstream components of p53-mediated apoptosis, suggesting that these genes play a role in radiation induced apoptosis. In this study, we transduced U-373MG cells harboring mutant p53 with the Apaf-1 and/or caspase-9 genes via adenoviral (Adv) vectors concomitant with X-ray irradiation and evaluated the degree of apoptosis. The percentage of apoptotic cells in U-373MG cells co-infected with the Adv for Apaf 1 (Adv-APAF-1) and that for caspase-9 (Adv-Casp9) and treated with irradiation (24%) was much higher than that in cells co-infected with Adv-APAF-1 and Adv Casp9 and not treated with irradiation (0.86%) and that in cells infected with either Adv-APAF-1 or Adv-Casp9 and treated with irradiation (2.0% or 2.6%, respectively). The apoptosis induced by co-transduction of Apaf-1 and caspase-9 and irradiation was repressed in cells that were co-infected with the Adv for Bcl X(L) but not in cells co-infected with the Adv for Bcl-2. These results indicate that Apaf-1 and caspase-9 play a role in radiation-induced apoptosis in cancer cells harboring mutant p53. Bcl-X(L) may be critically involved in the radioresistance of cancer cells by repressing Apaf-1- and caspase-9-mediated apoptosis. Expression of Apaf-1 and caspase-9 in tumors may be an important determinant of the therapeutic effect of irradiation in cancer treatment. PMID- 11410875 TI - IFN-gamma induces cell death in human hepatoma cells through a TRAIL/death receptor-mediated apoptotic pathway. AB - We demonstrated the induction of cell death in a hepatoma cell line by IFN-gamma and its possible mechanism. Among the 2 hepatitis B virus (HBV)-associated hepatoma cell lines, SNU-354 and SNU-368, IFN-gamma induced cell death and increased caspase-3 activity in SNU-368 but not in SNU-354. IFN-gamma induced several changes in the mRNA expression level of apoptosis-regulating genes, e.g., increased expression of Fas, caspase-1 and TNF-related apoptosis-inducing ligand (TRAIL). In particular, IFN-gamma potently increased the mRNA expression of TRAIL in both cell lines. However, it did not change the mRNA expression level of death mediating TRAIL receptors, e.g., DR4 and DR5, which were constitutively expressed in both cell lines. In contrast, the decoy receptor DcR1 was expressed in SNU-354 but not in SNU-368, and its expression level in SNU-354 was increased by IFN gamma. Another decoy receptor, DcR2, was constitutively expressed in both cell lines; however, its expression level in SNU-368 was decreased by IFN-gamma. In addition, exogenous recombinant TRAIL reduced viability in SNU-368, but not in SNU-354, cells. From these findings, we speculated that TRAIL up-regulation and the subsequent TRAIL-mediated apoptosis serve as a mechanism of IFN-gamma-induced cell death in SNU-368. To confirm this hypothesis, we demonstrated that soluble DR4-Fc fusion protein, a TRAIL pathway inhibitor, inhibited IFN-gamma-induced cell death in SNU-368. Our results demonstrated that IFN-gamma acts as an inducer of cell death through TRAIL-mediated apoptosis. PMID- 11410876 TI - Cancer treatment by photodynamic therapy combined with adoptive immunotherapy using genetically altered natural killer cell line. AB - Adoptive immunotherapy mediated by human natural killer (NK) cell line genetically altered to produce interleukin-2 (NK92MI) was evaluated as adjuvant to photodynamic therapy (PDT) of subcutaneous tumors. The combined effect of these two modalities was first examined with SiHa tumors (human cervical squamous cell carcinoma) growing in NOD-scid mice. The most effective protocol for NK92MI cell transfer in conjunction with PDT mediated by photosensitizer mTHPC was the injection of 5 x 10(7) cells (peritumoral or intravenous) given immediately after PDT, which produced a marked improvement in the therapeutic outcome compared with the effect of PDT alone. The same protocol was tested with HT-29 tumor model (human colorectal adenocarcinoma) xenografted in NOD-scid mice. The results demonstrate that the adoptive immunotherapy with NK92MI cells (which when used alone were not effective in controlling tumor growth) significantly improved the cures of PDT-treated HT-29 tumors, whereas such benefit was not observed with the parental cell line NK92 (not producing interleukin-2). Flow cytometry-based analysis revealed a higher percentage of p.t. injected NK92MI cells in PDT treated than in non-treated HT-29 tumors. Further investigation showed that the NK92MI cell-based adoptive immunotherapy is also a highly effective adjuvant for PDT treatment of murine EMT6 tumors growing in immunocompetent syngeneic BALB/c mice. This result diminishes the concern that adoptively transferred NK92MI cells may be rendered ineffective by an allogenic reaction of the host. The findings of this study suggest that advanced protocols of NK cell-based adoptive immunotherapy can be developed as efficient adjuvants to PDT used for the treatment of solid malignant tumors. PMID- 11410877 TI - Efficacy of antitumoral photodynamic therapy with hypericin: relationship between biodistribution and photodynamic effects in the RIF-1 mouse tumor model. AB - We investigated the hypericin-mediated PDT effects on the tumor and normal skin and in correlation with its biodistribution. These studies were carried out on C3H mice bearing RIF-1 tumors. The hypericin distribution and PDT effects were recorded at different intervals (0.5-24 hr) after intravenous injection of a 5 mg/kg dose of hypericin. After administration, rapid biphasic exponential decay was observed in the plasma drug concentration. It was found that hypericin was preferentially bound to the plasma lipoproteins. The tumor drug levels increased rapidly over the first few hours and reached a maximum around 6 hr after injection. In contrast, PDT efficacy was maximal when irradiation was performed at 0.5 hr after hypericin administration, which led to 100% cure. The PDT efficacy decreased rapidly as the administration-irradiation interval was prolonged. No tumor cure was obtained at the 6-hr interval, even though it was at this time that the tumor drug level peaked. Fluorescence microscopic studies showed that hypericin was mainly confined within the tumor vasculature at 0.5 hr after injection, whereupon it rapidly diffused to the surrounding tumor tissue. At 6 hr, a strong hypericin fluorescence was observed in the tumor tissue with only faint fluorescence within the vasculature, whereas at 24 hr the fluorescence in the tumor also decreased and became more diffused, and no fluorescence could be seen in the tumor vasculature. Like the tumor response, skin reactions were also found to be much more dramatic at short administration-irradiation intervals. Hypericin distribution and PDT response studies revealed a close correlation between the plasma drug level and the PDT effects, which suggests that vascular damage is the primary effect of hypericin-mediated PDT in this tumor model. PMID- 11410878 TI - Over-expression of cyclin A is highly associated with early relapse and reduced survival in patients with primary breast carcinomas. AB - Progression through the mammalian cell cycle is facilitated by cyclin-cyclin dependent kinase (cdk) complexes, which are activated at specific points during the cell cycle. Alteration in cyclin-cdk complexess may lead to altered cell cycle and tumorigenesis. In this study, we analyzed expression of cyclins A, D1, D3 and E in tumor tissue from 170 patients with primary invasive breast carcinomas. Immunohistochemical methods were used to detect protein expression of these cyclins. We detected positive immunoreactivity in 55 (32%), 22 (13%), 38 (22%) and 37 (21.8%) of the samples for cyclins A, D1, D3 and E, respectively. A highly statistically significant association was observed between expression of cyclin A and early relapse (p = 0.001 univariate analysis, p = 0.006 multivariate analysis) as well as cancer-specific death (p < 0.0001) during the follow-up time. No association was observed between cyclin D1 or cyclin E, respectively, and relapse of disease or survival, while cyclin D3 over-expression was associated with development of metastases during follow-up (p = 0.005 univariate analysis, p = 0.01 multivariate analysis). However, cyclin D3 did not show any statistically significant association when cancer-specific death was examined in a multivariate analysis (Cox regression for survival function). PMID- 11410879 TI - Cancer risk in heterozygotes for ataxia-telangiectasia. AB - Epidemiological studies have suggested that ataxia-telangiectasia (AT) heterozygotes have a predisposition to cancer, especially breast cancer in women. Now, haplotyping can identify heterozygotes for AT mutation (ATM) in AT families, allowing the risk of cancer associated with ATM heterozygosity status to be better assessed. We report a family study of AT patients, in which we estimated the risk of cancer according to ATM heterozygosity status. We analyzed demographic characteristics and occurrence of cancer in 1,423 relatives of AT patients. Haplotyping was performed in living relatives. The probability of being heterozygotes for ATM was calculated for deceased relatives. The risk of developing cancer was estimated in the cohort of relatives, and expected numbers of cancer cases were calculated from French age period-specific incidence rates. The number of cancers at all sites in the total population of relatives was not higher than expected. However, significant heterogeneity was found according to ATM heterozygosity status. This is mainly due to the increased risk of breast cancer previously observed in obligate heterozygotes. In obligate heterozygotes, relative risk (RR) was non-significantly increased for thyroid cancer, leukemia and liver cancer. Risks of ovarian, lung, pancreatic, kidney, stomach and colorectal cancers were non-significantly increased in the group with 0.5 probability of being heterozygotes. The RR was not significantly increased for any site of cancer, except for breast. Therefore, there is no evidence that specific screening of relatives of AT patients would be justified at particular sites other than the breast. However, the amplitude of the risk of breast cancer estimated in heterozygous women does not appear to justify a separate screening program from that already available to women with a first-degree relative affected by breast cancer. PMID- 11410880 TI - Second primary cancers after anogenital, skin, oral, esophageal and rectal cancers: etiological links? AB - The Swedish Family-Cancer Database was used to analyze second cancers after oral, esophageal, rectal, cervical, genital and skin (squamous cell carcinoma) cancers. A strong and consistent association of second cancers was observed at all these sites, in men and women. As a novel finding, an association of rectal cancer with the human papillomavirus (HVP)-related cancers was shown. New evidence on an excess of skin cancer with the HPV-related cancers was also provided. As an epidemiological study, the associations were strong and often supported by a number of comparisons. These could not be explained by bias or long-term treatment related effects. However, whether the findings on rectal and skin cancer are due to HPV or other infections, transient or inherited depressed immune function or other constitutional factors remains to be established. PMID- 11410881 TI - Alcoholism and risk for endometrial cancer. AB - Endogenous estrogens increase the risk of endometrial cancer and are also elevated among women with high alcoholic intake. It is incompletely known, however, whether alcohol intake in general and alcohol abuse in particular increases risk for endometrial cancer. We thus analyzed prospectively the risk for endometrial cancer among 36,856 women hospitalized with alcoholism between 1965 and 1994 through linkages between several national Swedish registers. Compared with the general population, women who were alcoholics had an overall 24% lower risk of developing endometrial cancer, a finding challenging our a priori hypothesis. However, among women below the age of 50 years at follow-up, the mean age of menopause among Swedish women, the risk was 70% higher, whereas the risk among women aged 50 years or more at follow-up was 40% lower compared with the general population. Hence, the effect of alcoholism on endometrial cancer appears to be age dependent. PMID- 11410883 TI - Adenoid cystic carcinoma: a retrospective clinical review. AB - Adenoid cystic carcinoma (ACC) are uncommon tumors, representing about 10% to 15% of head and neck tumors. We compare the survival and control rates at our institution with those reported in the literature, and examine putative predictors of outcome. All patients registered with the tumor registry as having had ACC were identified. Demographic and survival variables were retrieved from the database. Additionally, a chart review of all patients was done to obtain specific information. Minor gland tumors were staged using the American Joint Committee on Cancer's criteria for squamous cell carcinomas in identical sites. Histopathologic variables retrieved included grade of the tumor, margins, and perineural invasion. Treatment modalities, field sizes, and radiation doses were recorded in applicable cases. An effort to retrieve archival tumor specimens for immunohistochemical analysis was undertaken. A total of 69 patients were treated for ACC from 1955 to 1999. One patient, who presented with fatal brain metastasis, was excluded from further analysis. Of the remaining 68 patients, 30 were men and 38 were women. The average age at diagnosis was 52 years, and mean follow-up was 13.2 years. Mean survival was 7.7 years. Overall survival (OS) rates at 5, 10, and 15 years were 72%, 44%, and 34%, and cause-specific survival was 83%, 71%, and 55%, respectively. Recurrence-free survival rates were 65%, 52%, and 30% at 5, 10, and 15 years, with a total of 29 of 68 (43%) eventually suffering a recurrence. Overall survival was adversely affected by advancing T and AJCC stage. Higher tumor grades were also associated with decreased OS, although the numbers compared were small. Primaries of the nasosinal region fared poorly when compared with other locations. Total recurrence-free survival, local and distant recurrence rates were distinctly better in primaries of the oral cavity/oropharynx when compared with those in other locations. Reduced distant recurrence-free survival was significantly associated with increasing stage. No other variables were predictive for recurrence. Additionally, we found that nasosinal tumors were more likely to display higher stage at presentation, and were more often associated with perineural invasion. Also of interest was the association of perineural invasion with margin status, with 15 of 20 patients with positive margins displaying perineural invasion, while only 5 of 17 with negative margins showed nerve invasion (P = 0.02). On immunohistochemistry, 2 cases of the 29 (7%) tumor specimens found displayed HER-2/neu positivity. No correlation between clinical behavior and positive staining could be demonstrated. Our data concur with previous reports on ACC in terms of survival and recurrence statistics. Stage and site of primary were important determinants of outcome. Grade may still serve a role in decision making. We could not demonstrate any differences attributable to primary modality of therapy, perhaps due to the nonrandomization of patients into the various treatment tracks and the inclusion of palliative cases. Similarly, perineural invasion, radiation dose and field size, and HER-2/neu positivity did not prove to be important factors in our experience. PMID- 11410884 TI - Cyclin D1 overexpression associates with radiosensitivity in oral squamous cell carcinoma. AB - Overexpression of cyclin D1, a G1 cell cycle regulator, is often found in many different tumor types, including oral squamous cell carcinomas (SCC). Recent laboratory experiments have demonstrated that cyclin D1 levels can influence radiosensitivity in various cell lines. This study evaluated the relationship between cyclin D1 expression levels and radiosensitivity in nine oral SCC cell lines (HSC2, HSC3, HSC4, SCC15, SCC25, SCC66, SCC111, Ca9-22, and NAN2) and 41 clinical patients with oral SCC who underwent preoperative radiation therapy. Radiosensitivity of the nine oral SCC cell lines differed greatly in their response to radiation, assessed by a standard colony formation assay. Likewise, the expression of cyclin D1 varied, and the magnitude of the cyclin D1 expression correlated with increased tumor radiosensitivity. The similar significant association between the response to preoperative radiation therapy and cyclin D1 overexpression was observed in the oral SCC patients who were treated with preoperative radiation therapy. These results suggest that cyclin D1 expression levels correlate to radiosensitivity and could be used to predict the effectiveness of radiation therapy on oral SCC. PMID- 11410885 TI - Clonal chromosomal aberrations in simian virus 40-transfected human thyroid cells and in derived tumors developed after in vitro irradiation. AB - In vitro model cell systems are important tools for studying mechanisms of radiation-induced neoplastic transformation of human epithelial cells. In our study, the human thyroid epithelial cell line HTori-3 was analyzed cytogenetically following exposure to different doses of alpha- and gamma irradiation and subsequent tumor formation in athymic nude mice. Combining results from G-banding, comparative genomic hybridization, and spectral karyotyping, chromosome abnormalities could be depicted in the parental line HTori-3 and in nine different HTori lines established from the developed tumors. A number of chromosomal aberrations were found to be characteristic for simian virus 40 immortalization and/or radiation-induced transformation of human thyroid epithelial cells. Common chromosomal changes in cell lines originating from different irradiation experiments were loss of 8q23 and 13cen-q21 as well as gain of 1q32-qter and 2q11.2-q14.1. By comparison of chromosomal aberrations in cell lines exhibiting a different tumorigenic behavior, cytogenetic markers important for the tumorigenic process were studied. It appeared that deletions on chromosomes 9q32-q34 and 7q21-q31 as well as an increased copy number of chromosome 20 were important for the tumorigenic phenotype. A comparative breakpoint analysis of the marker chromosomes found and those observed in radiation-induced childhood thyroid tumors from Belarus revealed a coincidence for a number of chromosome bands. Thus, the data support the usefulness of the established cell system as an in vitro model to study important steps during radiation-induced malignant transformation in human thyroid cells. PMID- 11410886 TI - Ionizing radiation induces CD20 surface expression on human B cells. AB - The CD20 surface antigen is expressed on the great majority of B-cell lymphoma cases. The expression of this antigen ranges from moderate to bright, and it is neither internalized nor shed. These characteristics make CD20 a common target for antibody directed lymphoma therapy. The development of strategies to significantly increase CD20 expression on lymphoma cells is therefore of great interest as a means of increasing specific targeting and cell kill in antibody therapy and radio-immunotherapy. We present here data demonstrating that relatively low doses of external beam radiotherapy are capable of significant and consistent increases in CD20 surface expression in vitro. The effect is dose related up to approximately 10 Gy and is maximal in the first day after radiotherapy. We believe that these data may suggest a potent way to combine a short pretreatment course of external beam radiotherapy with a subsequent course of immunotherapy using either an unlabeled antibody or a radio-immunotoxin. PMID- 11410887 TI - Intra- and intertumor heterogeneity in blood perfusion of human cervical cancer before treatment and after radiotherapy. AB - Knowledge of the intratumor heterogeneity in blood perfusion may lead to increased understanding of tumor response to treatment. In the present work, absolute perfusion values, in units of ml/g.min, were determined in 20 tumor subregions of patients with cervical cancer before treatment (n = 12) and after 2 weeks of radiotherapy (n = 8), by using a method based on contrast-enhanced magnetic resonance imaging. The aims were to evaluate the intratumor heterogeneity in perfusion in relation to the intertumor heterogeneity and to search for changes in the heterogeneities during the early phase of therapy. The intra- and intertumor heterogeneity in perfusion were estimated from components of one-way analyses of variance. The mean perfusion differed significantly among the patients before treatment, ranging from 0.044 to 0.12 ml/g x min. Large differences in perfusion were also observed within individual tumors. The heterogeneity was largest in the best perfused tumors, perfusion values ranging, e.g., from 0.055 to 0.29 ml/g x min were observed. The intratumor heterogeneity was similar to the intertumor heterogeneity. The mean perfusion generally increased or tended to increase during radiotherapy, ranging from 0.064 ml/g x min to 0.13 ml/g x min after 2 weeks of treatment. There was a tendency of increased intratumor heterogeneity in perfusion after therapy, consistent with the higher mean value; a difference in perfusion of more than a factor of 10 was seen within some tumors. These results suggest that cervix tumors contain a significant amount of poorly perfused subregions with high treatment resistance. Moreover, the perfusion and perfusion heterogeneity may increase during the early phase of radiotherapy and influence tumor response. PMID- 11410888 TI - The sensitivity and specificity of FDG PET in distinguishing recurrent brain tumor from radionecrosis in patients treated with stereotactic radiosurgery. AB - Radiation necrosis and recurrent brain tumor have similar symptoms and are indistinguishable on both magnetic resonance imaging (MRI) and computed tomograph scans. 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) has been proposed as a diagnostic alternative, particularly when co-registered with MRI. We studied 47 patients with brain tumors treated with stereotactic radiosurgery and followed with FDG PET. For all tumor types, the sensitivity of FDG PET for diagnosing tumor was 75% and the specificity was 81%. For brain metastasis without MRI co-registration, FDG PET had a sensitivity of 65% and a specificity of 80%. For brain metastasis with MRI co-registration, FDG PET had a sensitivity of 86% and specificity of 80%. MRI co-registration appears to improve the sensitivity of FDG PET, making it a useful modality to distinguish between radiation necrosis and recurrent brain metastasis. PMID- 11410889 TI - Clinical staging of prostate cancer: reproducibility and clarification of issues. AB - The American Joint Committee on Cancer (AJCC) staging system for prostate cancer adopted in 1992 is based on tumor-node-metastasis (TNM) designations. It has been widely accepted for use in local and advanced disease. The purpose of this study was to assess reproducibility of staging among observers and to help clarify staging issues. Twelve prostate cancer cases were sent to 20 physicians with special expertise in prostate cancer including eight urologists, eight radiation oncologists, and four medical oncologists. Physicians were asked to assign a stage based on the 1992 AJCC clinical staging. The most frequently reported stage assigned to each case was taken to be the consensus. Agreement was the percentage of physicians who reported that particular stage. Seventy-five percent of the physicians responded. The overall agreement for assignment of T stage was 63.9%. Differences were found by specialty for inclusion of available information in designating a T stage. The overall agreement for N stage was 73.8%. The most common designation was Nx regardless of availability of a computed tomography scan. The overall agreement for M stage was 76.6%. Without a bone scan the most common designation was Mx regardless of Gleason grade or prostate-specific antigen (PSA). A frequent comment was that PSA was more indicative of disease extent than current clinical staging. The reproducibility of the 1992 clinical AJCC staging is poor even among experts in the field. This problem arises primarily from disagreement regarding which studies are included in assigning a stage. Some of these difficulties are addressed in the 1997 revision. However, the clinical staging does not address the true biological significance of disease in many instances. PMID- 11410890 TI - Dynamic mechanical characterization of biodegradable composites of hydroxyapatite and polylactides. AB - The thermo-mechanical analysis of some novel, totally bioactive, three-component composite systems intended for the repair and augmentation of bone has been performed. The composites are composed of hydroxyapatite (HA), particulate reinforced biodegradable polylactic acid (PDLLA), with a second reinforcing phase of semi-crystalline, unidirectional fibers of polylactic acid (PLLA) homopolymer. Dynamic mechanical analysis (DMA) applied small-strain cyclic deformations to simulate physiological loading. The effect of the two fillers on the glass transition temperature of the matrix (Tg-PDLLA) and on the viscoelastic parameters, storage modulus (EI), and tan delta were analyzed. Thermal characterization with differential scanning calorimetry (DSC) was compared to the DMA data. It was established that the addition of HA and/or PLLA fibers increased EI, both below and above Tg-PDLLA. The combination of both reinforcing agents was the most effective. The damping was reduced with the addition of HA alone and in combination with PLLA fibers. Tg-PDLLA was increased by the addition of HA, PLLA fibers, and the combination of the two reinforcing agents. The magnitude of the increase in Tg-PDLLA was similar for both types of reinforcement. PMID- 11410891 TI - Development of the shields for tendon injury repair using polyvinyl alcohol- hydrogel ( PVA-H). AB - In recent years, marked advances have been made in repair techniques for tendon injury, but the treatment of finger flexor tendon injury is still one of the most difficult and important problems in the orthopedic field. The main problem in tendon repair is adhesion between the tendon and surrounding tissue. To prevent this adhesion and achieve tendon union, we developed adhesion preventive shields for tendon repair using polyvinyl alcohol hydrogel ( PVA-H) with 90% water content, and carried out an implant experiment using the deep flexor tendon of the third toe of domestic fowl. Injured tendons shielded with PVA-H showed union at about 3 weeks after the operation without adhesion to the surrounding tissue and good function such as gliding and range of motion. Neither breakage of the PVA-H shield itself nor infection, nor degeneration in the surrounding tissue were observed. These results confirmed that the tendon itself has repair ability, and the tendon is regenerated by synovial nutrition through PVA-H. High water content PVA-H may have clinically potential and be applicable to adhesion preventive shields for tendon repair. However, re-rupture was observed, probably due to accidental tendon injury at an early period after the operation. In some cases, tendon immobilization methods to prevent re-rupture might be necessary. PMID- 11410892 TI - Control of pore structure and size in freeze-dried collagen sponges. AB - Because of many suitable properties, collagen sponges are used as an acellular implant or a biomaterial in the field of tissue engineering. Generally, the inner three-dimensional structure of the sponges influences the behavior of cells. To investigate this influence, it is necessary to develop a process to produce sponges with a defined, adjustable, and homogeneous pore structure. Collagen sponges can be produced by freeze-drying of collagen suspensions. The pore structure of the freeze-dried sponges mirrors the ice-crystal morphology after freezing. In industrial production, the collagen suspensions are solidified under time- and space-dependent freezing conditions, resulting in an inhomogeneous pore structure. In this investigation, unidirectional solidification was applied during the freezing process to produce collagen sponges with a homogeneous pore structure. Using this technique the entire sample can be solidified under thermally constant freezing conditions. The ice-crystal morphology and size can be adjusted by varying the solute concentration in the collagen suspension. Collagen sponges with a very uniform and defined pore structure can be produced. Furthermore, the pore size can be adjusted between 20-40 microm. The thickness of the sponges prepared during this research was 10 mm. PMID- 11410893 TI - Chitosan-alginate films prepared with chitosans of different molecular weights. AB - Chitosan-alginate polyelectrolyte complex (CS-AL PEC) is water insoluble and more effective in limiting the release of encapsulated materials compared to chitosan or alginate. Coherent CS-AL PEC films have been prepared in our laboratory by casting and drying suspensions of chitosan-alginate coacervates. The objective of this study was to evaluate the properties of the CS-AL PEC films prepared with chitosans of different molecular weights. Films prepared with low-molecular weight chitosan (Mv 1.30 x 10(5)) were twice as thin and transparent, as well as 55% less permeable to water vapor, compared to films prepared with high-molecular weight chitosan (Mv 10.0 x 10(5)). It may be inferred that the low-molecular weight chitosan reacted more completely with the sodium alginate (M(v) 1.04 x 10(5)) than chitosan of higher molecular weight. A threshold molecular weight may be required, because chitosans of Mv 10.0 x 10(5) and 5.33 x 10(5) yielded films with similar physical properties. The PEC films exhibited different surface properties from the parent films, and contained a higher degree of chain alignment with the possible formation of new crystal types. The PEC films exhibited good in vitro biocompatibility with mouse and human fibroblasts, suggesting that they can be further explored for biomedical applications. PMID- 11410894 TI - Micromechanics of the dentin/adhesive interface. AB - Scanning acoustic microscopy (SAM) was used in the burst mode at 400 MHz, nominal lateral resolution 2.5 microm, to study the micromechanical properties of the dentin/adhesive interface. Corresponding specimens from the same tooth were investigated using mu Raman spectroscopy, light microscopy, and scanning electron microscopy.12 The elastic moduli of the components of the dentin/adhesive interface were determined by comparing the recorded acoustic impedance values to a calibration curve generated on standard materials. The standard materials, which include polypropylene, Teflon, PMMA, pyrex glass, aluminum, titanium, and stainless steel, provide the appropriate range of acoustic impedance values. The elastic moduli of the components of the dentin/adhesive interface are: partially demineralized dentin, 13 Gpa; mineralized dentin, 28 GPa; adhesive, 5.0 GPa; and unprotected protein at the interface < 2.0 GPa. PMID- 11410895 TI - Blends of stearyl poly(ethylene oxide) coupling-polymer in chitosan as coating materials for polyurethane intravascular catheters. AB - To optimize the surface biocompatibility of the intravascular catheter, an amphiphilic coupling-polymer of stearyl poly (ethylene oxide) -co- 4,4'-methylene diphenyl diisocyanate-co- stearyl poly (ethylene oxide), for short MSPEO, was specially designed as the surface modifying additive (SMA). The blend of MSPEO in chitosan was coated on the outer wall of the catheters by the dip-coating method. The surface analysis was carried out by ATR-FTIR and contact angle measurements. The surface enrichment of MSPEO was confirmed. On the water interface, the larger the molecular weight of PEO was, the higher the surface enrichment. While on air interface, the case was the contrary. Three kinds of static test of clotting time, plasma recalcification time (PRT), prothrombin time (PT), and thrombin time (TT), as well as the static platelet adhesion experiment were carried out. The results indicated that the coated surface could resist the clotting effectively. In order to test the blood-compatibility of the coated catheters under a shear of blood flow, the dynamic experiment was performed through a closed-loop tubular system with the shear rate of 1500 s(-1). The results of blood regular testing at six different times (0, 5,10, 20, 30, and 60 min) indicated that the biocompatibility of the coating was nearly ideal. Finally, the SMA-MSPEO was proved to be non-chronic-toxic by animal experiments with rats and suitable as a coating material for clinical use. PMID- 11410896 TI - Animal study on surface-modified defibrillator systems: Indications for enhanced infection resistance. AB - One of the most important problems with ICD systems is infection. The aim of this study was an in vivo evaluation of the efficacy of defibrillator systems in terms of infection resistance. The polyurethane leads were coupled with heparin and loaded with the antibiotic gentamicin, while the PGs were modified to release gentamicin. Group I was comprised of 10 pigs implanted with either a standard or a modified system for 2 weeks; group II was implanted during 4 weeks. The lead was inserted into the heart wall via the jugular vein. The other end was subcutaneously tunneled to the armpit where the PG was positioned. A cocktail of Staphylococcus aureus and epidermidis was injected at the site of the PG. Evaluation was performed macroscopically, by taking bacterial swabs during explantation and by microscopic processing. The results showed that 3 out of 5 modified defibrillator-systems in group I and 1-2 out of 5 in group II were judged as noninfected, whereas all standard systems were infected. Infection rates of the remaining modified defibrillators showed variances, as found with the standards, from slight to moderate to high, to even high/severe in group II (1x standard and 1x modified). With the modified systems, this may be related to production of humoral factors by an intensified early tissue reaction, as indicated by a swelling at day 6 at the site of the PG. When infected, whether or not modified, usually only Staphylococcus aureus was present. Spreading of infection seemed to occur by inoculation via blood, for example, based on the observation that group II in general showed an increase in infected fibrotic overgrowth in the heart, while infectious problems were low in the jugular vein. It is concluded that the modification at short term shows enhanced infection resistance. An increased infection rate already at 4 weeks, however, indicates that the modification may not hold in the long run. Special attention is needed concerning the more intense early tissue reaction. PMID- 11410897 TI - Osteoprotegerin inhibits in vitro mouse osteoclast formation induced by joint fluid from failed total hip arthroplasty. AB - Osteoprotegerin (OPG) is a key regulator of osteoclastogenesis. We investigated the presence of OPG and bone-resorbing cytokines, the potential of osteoclastic differentiation in joint fluid from failed total hip arthroplasty (THA), and the inhibitory effect of OPG on osteoclast formation in vitro induced by the joint fluid. The study was aimed to clarify one important step in the cascade of periprosthetic osteolysis in the process of implant loosening. OPG levels in failed THA joint fluid of 20 cases were significantly lower than osteoarthritis (OA) joint fluid of 15 cases (p < 0.001). The levels of bone-resorbing cytokines, interleukin (IL)-1beta, and IL-6 were significantly higher in failed THA joint fluid than OA fluid (p < 0.001 and p = 0.001, respectively). Marked osteoclast formation was observed in the presence of failed THA joint fluid in the mouse coculture system, when compared to OA fluid (p < 0.001). The addition of 100 ng/mL OPG to the mouse coculture system completely inhibited osteoclast formation in the presence of failed THA joint fluid (p < 0.001). The data suggest that low levels of OPG combined with higher IL-1beta and IL-6 levels represent the potential of osteoclast differentiation and its activation in failed THA joint fluid. Inhibition of osteoclastogenesis in vitro by OPG suggests that a low level of OPG with elevated bone resorbing cytokines contributes to periprosthetic osteolysis via osteolytic joint fluid, thus leading to THA prosthesis loosening. PMID- 11410898 TI - New pulsatile bioreactor for fabrication of tissue-engineered patches. AB - To date, one approach to tissue engineering has been to develop in vitro conditions to ultimately fabricate functional cardiovascular structures prior to final implantation. In our current experiment, we developed a new pulsatile flow system that provides biochemical and biomechanical signals to regulate autologous patch-tissue development in vitro. The newly developed patch bioreactor is made of Plexiglas and is completely transparent (Mediport Kardiotechnik, Berlin). The bioreactor is connected to an air-driven respirator pump, and the cell culture medium continuously circulates through a closed-loop system. We thus developed a closed-loop, perfused bioreactor for long-term patch-tissue conditioning, which combines continuous, pulsatile perfusion and mechanical stimulation by periodically stretching the tissue-engineered patch constructs. By adjusting the stroke volume, the stroke rate, and the inspiration/expiration time of the ventilator, it allows various pulsatile flows and different levels of pressure. The whole system is a highly isolated cell culture setting, which provides a high level of sterility, gas supply, and fits into a standard humidified incubator. The bioreactor can be sterilized by ethylene oxide and assembled with a standard screwdriver. Our newly developed bioreactor provides optimal biomechanical and biodynamical stimuli for controlled tissue development and in vitro conditioning of an autologous tissue-engineered patch. PMID- 11410899 TI - Inhibition of leukotriene function can modulate particulate-induced changes in bone cell differentiation and activity. AB - Aseptic loosening remains the major problem facing arthroplasty longevity with particulates from component materials touted as the cause of periprosthetic osteolysis. Proposed mechanisms in aseptic bone loss include: increased resorption, increased differentiation of osteoclasts (and/or macrophages locally), and decreased osteoblastic bone formation. Leukotrienes participate in osteoclastic bone resorption. We investigated inhibiting leukotrienes synthesis, using ICI 230487, to ameliorate the effects of particulates on osteoclast pit formation and also assessed the effects of alendronate, a bisphosphonate, on pit formation. Three particulates were used: ultra high molecular weight polyethylene (UHMWPE), polymethylmethacrylate (PMMA) and hydroxyapatite (HA). Osteoclast resorption was increased with UHMWPE, PMMA, and HA particles. Interventions with alendronate and ICI 230487 reduced particulate-induced osteoclast resorption. Both ICI 230487 and alendronate reduced osteoclast numbers at higher doses. To assess the effect of particulates on osteoclast and macrophage differentiation, mouse bone marrow was cultured and stained for tartrate resistant acid phosphatase colonies (TRAP+, osteoclasts) and nonspecific esterase positive colonies (NSE+, macrophage precursors). Particulates increased both TRAP+ and NSE+ colony formation. These increases were inhibited by ICI 230487. Particulates also inhibited osteoblast function assessed by the development of mineralized nodules and alkaline phosphatase positive (AP+) colony area. ICI 230487 partly protected osteoblast function from this particulate effect. Blockade of leukotriene production may prove a useful therapeutic intervention for particulate-induced aseptic loosening by inhibiting resorptive activity, reducing the pro-inflammatory cell populations induced and recruited by these particulates, as well as ameliorating the negative effects of inflammatory mediators on osteoblast function. PMID- 11410900 TI - Quantitative characterization of polyethylene debris isolated from periprosthetic tissue in early failure knee implants and early and late failure Charnley hip implants. AB - This study isolated and characterized UHMWPE particles from 3 explant groups: early Charnley hip failures (ECE; < 10 years), late Charnley hip failures (LCE; > 10 years) and early knee failures (EKE; < 10 years). Debris isolated from the 3 groups had percentage particle number and percentage volumetric concentration distributions that were not significantly different. The greatest number of particles were found in the 0.1-0.5 microm size range and 19-20.6% of the volumetric concentration was below 1 microm in size in all groups. However, there were significant differences in the total volumetric concentration of debris isolated per g of tissue. LCE had significantly higher volumes of debris than ECE and EKE, there was no significant difference in the volume of debris from the EKE and ECE. The mean aspect ratio and mean irregularity ratio of the LCE group were also significantly higher than the ECE and EKE, suggesting that different wear mechanisms were occurring in the late Charnley group compared to the early Charnley and knee groups. These results also suggest that early knees, with normal surface wear, may have similar wear mechanisms to early Charnley hips and indicate that similar volumes of biologically active micrometer and sub micrometer UHMWPE particles were produced. This may have important implications in the longer-term outcome of total knee arthroplasties, because it indicates a similar potential for osteolysis induced by wear debris. PMID- 11410901 TI - Air-flow resistances of silicone rubber voice prostheses after formation of bacterial and fungal biofilms. AB - Laryngectomized patients use silicone rubber voice prostheses to rehabilitate their voice. However, biofilm formation limits the lifetime of voice prostheses by causing leakage or an increased air-flow resistance and the prosthesis has to be replaced. To determine which bacterial or yeast strains, isolated from explanted voice prostheses, contribute most to increases in air-flow resistance of silicone rubber voice prostheses, biofilms consisting of either a bacterial or a yeast strain were grown on voice prostheses in the artificial throat model. The effects of these biofilms on air-flow resistances were determined by calculating the difference in air-flow resistance of the individual voice prosthesis as covered with a 7-day-old biofilm with the situation prior to biofilm formation. Conspicuously, voice prosthetic biofilms formed by the bacterial strains Staphylococcus aureus GB 2/1 and Rothia dentocariosa GBJ 41/25B and their excreted organic matter showed larger increases in air-flow resistance (more then 30 cm H(2)O.s/L) than biofilms formed by Candida species. This is contrary to the literature, where there seems to be agreement that Candida species are mainly responsible for clinical failure of silicone rubber voice prostheses. PMID- 11410902 TI - Physicochemical, mechanical, and biological properties of commercial membranes for GTR. AB - Barrier membranes for guided tissue regeneration (GTR) to treat bone defects have to satisfy criteria of biocompatibility, cell-occlusiveness, spacemaking, tissue integration, and clinical manageability. In this study, the morphological and mechanical properties of two commercial biodegradable membranes (Resolut LT and Biofix) as a function of the incubation time have been compared. Moreover, their permeability to both fluids and epithelial cells as well as the bacteria adhesion have been evaluated. The membranes are asymmetric and composed of a dense polymeric layer coupled with nonwoven (Resolut LT) or woven (Biofix) fibers. Both of the membranes, when incubated in complete culture medium, completely lose the structural and mechanical properties within 30 days. Moreover the results of solute permeability show that Resolut LT and Biofix membranes cannot be considered selective membranes to the solute crossing. On the contrary, they act as a barrier to the passage of the gingivial cells and to S. mutans bacteria. PMID- 11410903 TI - Quantitative evaluation of the prosthetic head damage induced by microscopic third-body particles in total hip replacement. AB - The increase of the femoral head roughness in artificial hip joints is strongly influenced by the presence of abrasive particulate entrapped between the articulating surfaces. The aim of the present study is to evaluate the dependence of such damage on the geometry of the particles entrapped in the joint, with reference to the UHMWPE/chrome-cobalt coupling. Five chrome-cobalt femoral heads and their coupled UHMWPE acetabular cups, retrieved at revision surgery after a short period of in situ functioning, have been investigated for the occurrence of third-body damage. This was found on all the prosthetic heads, where the peak-to valley height of the scratches, as derived from profilometry evaluations, ranged from 0.3-1.3 microm. The observed damage has been divided into four classes, related to the particle motion while being embedded into the polymer. Two kinds of particle morphology have been studied, spherical and prismatic, with size ranging from 5-50 microm. In order to provide an estimation of the damage induced by such particles, a finite element model of the third-body interaction was set up. The peak-to-valley height of the impression due to the particle indentation on the chrome-cobalt surface is assumed as an index of the induced damage. The calculated values range from 0.1-0.5 microm for spherical particles of size ranging from 10-40 microm. In the case of prismatic particles, the peak-to-valley height can reach 1.3 microm and depends both on the size and width of the particle's free corner, indenting the chrome-cobalt. As an example, a sharp-edged particle of size 30 microm can induce on the chrome-cobalt an impression with peak-to-valley height of 0.75 microm, when embedded into the polyethylene with a free edge of 5 microm facing the metallic surface. Negligible damage is induced, if a free edge of 7.5 microm is indenting the counterface. Our findings offer new support to the hypothesis that microscopic third-body particles are capable of causing increased roughening of the femoral head and provide a quantitative evaluation of the phenomenon. PMID- 11410904 TI - Biodegradable poly(D,L-lactide) coating of implants for continuous release of growth factors. AB - Local application of growth factors like insulin like growth factor-I (IGF-I) and transforming growth factor-beta 1 (TGF-beta1) from a biodegradable thin layer of poly(D,L-lactide) (PDLLA) coated implants could stimulate fracture healing. A new "cold coating technique" for metallic implants was established to produce a biodegradable coating with a high mechanical stability that provides a continuous release of incorporated growth factors. The properties of this bioactive coating were investigated in vitro and in vivo. Scanning electron microscope analysis revealed a coating thickness of in average 14.8 microm on titanium and 10.7 microm on steel wires. Intramedullary implantation and extraction experiments depicted a loss of PDLLA coating from titanium and steel implants of less than 5%. After explantation of the implants, the coating displayed a complete and regular layer without any defects of PDLLA uncovering the metallic surface. Smear tests demonstrate that the coating can be performed under sterile conditions. The PDLLA depicted a reduction of about 8% within 6 weeks in vitro and in vivo. The growth factors were incorporated in a stable form and demonstrated a loss of stability of less than 3% within 42 days and less than 5% within one year. In an elution experiment, 54% IGF-I and 48% TGF-beta1 were released within the first 48 h. After 42 days, 76% of IGF-I and 71% of TGF-beta1 were detected in the elution fluid by ELISA. Comparable results were obtained in the in vivo experiments after 42 days. PMID- 11410905 TI - Development of PMMA-precoating metal prostheses via injection molding: residual stresses. AB - Injection molding polymethylmethacrylate (PMMA) was developed as a potential metal prosthesis precoating method for surgical implantation. However, residual stresses generated in the injection-molded PMMA affect the mechanical performance of the coating polymer. This study used plane polariscope testing to demonstrate the residual stress distribution in the injection-molded polymer. Slitting and strain gages were combined with finite element analysis to give a quantitative measure of the residual stresses. The residual stress testing indicated that the highest circumferential residual stress in the injection-molded PMMA was approximately 5 MPa in tension. Two methods, water soaking and slitting, were developed for reducing residual stresses. A series of four surgical implantation simulation processes were designed. The two methods for reducing residual stress were used in the simulations. PMID- 11410906 TI - Paternal chemical exposures, hormone levels, and offspring sex ratios: comment on Trasler ('00). PMID- 11410908 TI - Geometric morphometrics of corpus callosum and subcortical structures in the fetal-alcohol-affected brain. AB - BACKGROUND: Although experienced clinicians have been diagnosing fetal alcohol syndrome (FAS) for nearly 30 years, the rest of the spectrum of fetal alcohol damage is not being classified effectively. This article describes a quantification of neuroanatomical structure that may supply a useful discriminator of prenatal brain damage from alcohol. It is demonstrated in a data set of adults of both sexes. METHODS: Ninety adults (45 males) were examined by magnetic resonance imaging (MRI). These subjects were group-matched for age and ethnicity across three diagnoses: FAS, fetal alcohol effects (FAE), and normals. All FAS and FAE were heavily alcohol-exposed in utero; normals were not. From T(1)-weighted MR brain images, we extracted 3D morphometric representations of shape for 33-landmark point configurations and 40-point outlines of the corpus callosum along its midline (a slightly nonplanar structure). RESULTS: There are striking differences between exposed and unexposed in the statistical distributions of these two shapes. The differences are better characterized by excess variance in the exposed group than by any change in average landmark or outline shape. For each sex, combining the callosal outline data with the landmark data leads to a powerful quadratic discriminator of exposed from unexposed. The discriminating features include the relationship of brain stem to diencephalon, and localized variabilities of callosal outline shape, but not diagnosis (FAS vs. FAE). CONCLUSIONS: Statistical analysis of brain shape is a powerful new source of information relevant to fetal alcohol spectrum nosology and etiology. Patients with FAS and FAE do not differ in these brain shape features, but both differ from the unexposed. The aspects of brain shape that are especially variable may be entailed in the underlying neuroteratogenetic mechanisms. PMID- 11410909 TI - Vitamins C and E improve rat embryonic antioxidant defense mechanism in diabetic culture medium. AB - BACKGROUND: Diabetes teratogenicity seems to be related to embryonic oxidative stress and the extent of the embryonic damage can apparently be reduced by antioxidants. We have studied the mechanism by which antioxidants, such as vitamins C and E, reduce diabetes-induced embryonic damage. We therefore compared the antioxidant capacity of 10.5-day-old rat embryos and their yolk sacs cultured for 28h in diabetic culture medium with or without vitamins C and E. METHODS: The embryos were cultured in 90% rat serum to which 2mg/ml glucose, 2mg/ml beta hydroxy butyrate (BHOB) and 10 microg/ml of acetoacetate were added. Rat embryos were also cultured in a diabetic medium with 25 microg/ml of vitamin E and 50 microg/ml of vitamin C. Control embryos were cultured in normal rat serum with or without vitamins C and E. RESULTS: Decreased activity of Cu/Zn superoxide dismutase (SOD) and of catalase (CAT) in the "diabetic" embryos and their yolk sacs, and reduced concentrations of low molecular weight antioxidant (LMWA) were found. Under these conditions we also found a decrease in vitamin C and vitamin E concentrations in the embryos, as measured by HPLC. In situ hybridization for SOD mRNA showed a marked reduction of SOD mRNA in the brain, spinal cord, heart and liver of embryos cultured in diabetic medium in comparison to controls. Following the addition of vitamins C and E to the diabetic culture medium, SOD and CAT activity, the concentrations of LMWA, the levels of vitamin C and E and the expression of SOD mRNA in the embryos and yolk sacs returned to normal. CONCLUSIONS: Diabetic metabolic factors seem to have a direct effect on embryonic SOD gene and perhaps genes of other antioxidant enzymes, reducing embryonic endogenous antioxidant defense mechanism. This in turn may cause a depletion of the LMWA, such as vitamins C and E. The addition of these vitamins normalizes the embryonic antioxidant defense mechanism, reducing the damage caused by the diabetic environment. PMID- 11410910 TI - Effect of early fetal irradiation on the postnatal development of mouse. AB - BACKGROUND: An earlier study has shown that irradiation at the late fetal stage of Swiss albino mice disturbed postnatal growth and appearance of physiological markers. The present study was done to determine the effect of irradiation at the early fetal stage on the postnatal development of mouse. METHODS: Fourteen-day pregnant Swiss albino mice were exposed to 0.1-1.5 Gy of cobalt-60 gamma-rays. F1 pups were observed for < or = 6 weeks of age. Postnatal mortality, body weight and length, head length and width, tail length, and the time of appearance of physiological markers (pinna detachment, eye opening, fur development, vaginal opening and testes descent) were noted. RESULTS: There was no increase in congenital anomalies. Postnatal mortality and percentage of growth-retarded pups increased significantly at doses of 0.5-1.5 Gy. A significant delay in the appearance of all the physiological markers was also noted at these doses. Body length, head length, and tail length remained significantly lower than in the controls throughout the observation period at doses of 0.3-1.5 Gy, whereas body weight and head width showed such a persistent change only at > or = 0.5 Gy. CONCLUSIONS: The early fetal day 14 in mouse is sensitive to radiation-induced postnatal mortality and impairment of growth and temporal development of physiological markers, but not to induction of congenital anomalies. While mortality and physiological markers are not affected at <0.5 Gy, growth retardation appears to have a lower threshold of approximately 0.3 Gy. PMID- 11410911 TI - Teratogen update: evaluation of the reproductive and developmental risks of caffeine. AB - Caffeine is a methylated xanthine that acts as a mild central nervous system stimulant. It is present in many beverages, including coffee, tea, and colas, as well as chocolate. Caffeine constitutes 1-2% of roasted coffee beans, 3.5% of fresh tea leaves, and approximately 2% of mate leaves (Spiller, '84; Graham, '84a,b). Many over-the-counter medications, such as cold and allergy tablets, headache medicines, diuretics, and stimulants also contain caffeine, although they lead to relatively minimal intake (FDA, '86). In epidemiological studies, it is assumed that one cup of coffee contains < or =100 mg of caffeine, and soft drinks, such as colas, contain 10-50 mg of caffeine per 12-ounce serving. The per capita consumption of caffeine from all sources is estimated to be about 3-7 mg/kg per day, or approximately 200 mg/day (Barone and Roberts, '96). Consumption of caffeinated beverages during pregnancy is quite common (Hill et al., '77) and is estimated to be approximately 144 mg/day, or 2.4 mg/kg for a 60-kg human (Morris and Weinstein, '81). However, pregnant women appear to consume slightly less than do other adults, approximately 1 mg/kg per day (Barone and Roberts, '96). This decrease may be interrelated with taste aversion (Hook, '76; Little, '82). The medical literature contains many varied references that appear to indicate that human adverse reproductive/developmental effects are produced by caffeine. If caffeine indeed causes such effects, the reproductive consequences could be very serious because caffeine-containing foods and beverages are consumed by most of the human populations of the world, and consumption in the United States is estimated to be 4.5-kg/person/year (Narod et al., '91). Therefore, the medical literature dealing with developmental and reproductive risks of caffeine was reviewed, and the biological plausibility of the epidemiological and animal findings, as well as the methods and conclusions of previous investigators, were evaluated. The epidemiological studies describe exposures of women to caffeine during pregnancy, as well as the occurrence of congenital malformations, fetal growth retardation, small-for-date babies, miscarriages (spontaneous abortions), behavioral effects, and maternal fertility problems that presumably resulted from the caffeine consumption. A few epidemiological studies were concerned with the genetic effects of preconception exposures to caffeine. Animal studies, conducted mostly in pregnant rats and mice, were designed to produce malformations. The objectives of the present review are to summarize the findings from the various clinical and animals studies, objectively discuss the merits and/or faults inherent in the studies and establish a global reproductive risk assessment for caffeine consumption in humans during pregnancy. It should be noted that evaluation of the developmental risks of caffeine based solely on epidemiological studies is difficult because the findings are inconsistent. Even more important, is the fact that caffeine users are subject to multiple confounding factors that make analyses difficult and prevent investigators from reaching definitive conclusions. For example, the caffeine content of foods and beverages can vary considerably, which can interfere with obtaining valid interpretations from many human studies. Isolated epidemiological studies dealing with the risk of abortion, without evaluating other developmental and reproductive effects, are the most difficult to interpret, because they present special problems that are sometimes ignored in epidemiological studies. The results of animal studies are probably most helpful in solving some of the dilemmas created by the epidemiological studies. An animal study reported in 1960 first focused our attention on the potential developmental effects of caffeine. However, the exposure reported by Nishimura and Nakai ('60) was an intraperitoneal dosage of 250 mg/kg in the mouse, an extremely high dosage that would result in a blood plasma level that could never be obtained from consuming caffeine containing products. More recent animal studies have demonstrated, that depending on the method of administration and species, the developmental NOEL in rodents is approximately 30 mg/kg per day, the teratogenic NOEL is 8,100 mg/kg per day, and the reproductive NOEL approximately 80-120 mg/kg per day. Lack of biological plausibility to support the concept that caffeine has been responsible for human malformations is another important part of this analysis. For example, no one has described the Caffeine "teratogenic syndrome," a cluster of malformations associated with caffeine ingestion. Proven human teratogens have an identifiable syndrome. The malformations described in the animal studies at very high doses fit the description of vascular disruptive types of malformations. (ABSTRACT TRUNCATED) PMID- 11410912 TI - Excitability testing in clinical neurophysiology--what, why, and when? PMID- 11410913 TI - Role of motor unit structure in defining function. AB - Motor units, defined as a motoneuron and all of its associated muscle fibers, are the basic functional units of skeletal muscle. Their activity represents the final output of the central nervous system, and their role in motor control has been widely studied. However, there has been relatively little work focused on the mechanical significance of recruiting variable numbers of motor units during different motor tasks. This review focuses on factors ranging from molecular to macroanatomical components that influence the mechanical output of a motor unit in the context of the whole muscle. These factors range from the mechanical properties of different muscle fiber types to the unique morphology of the muscle fibers constituting a motor unit of a given type and to the arrangement of those motor unit fibers in three dimensions within the muscle. We suggest that as a result of the integration of multiple levels of structural and physiological levels of organization, unique mechanical properties of motor units are likely to emerge. PMID- 11410914 TI - Effects of temperature on neuromuscular electrophysiology. AB - Like nearly all biologic structures, the peripheral nervous system is remarkably temperature sensitive. Clinical neurophysiologists are most aware of the untoward effects of cooling on nerve conduction studies, including reduced conduction velocity, prolonged distal latency, and increased response amplitude and duration. However, familiarity with the effects of temperature variation on the peripheral nervous system can also provide a deeper understanding of the physiological mechanisms underlying the function of nerve, muscle, and neuromuscular junction in health and disease. Intentional temperature alteration can also improve the diagnostic accuracy of certain electrophysiologic tests, such as the use of heat when performing repetitive nerve stimulation in myasthenia gravis or the use of cold during needle electromyography in some of the myotonic disorders. Finally, extremes of temperature have long been known to produce permanent neuronal dysfunction; recent investigations are beginning to elucidate the mechanisms of such injury. PMID- 11410915 TI - Clinical evaluation of excitability measures in sensory nerve. AB - A recently described method for recording multiple excitability parameters of human motor nerves has been adapted to the study of sensory nerves. The protocol measures stimulus-response behavior using two stimulus durations (from which the distribution of strength-duration time constants is estimated), threshold electrotonus to 100 ms polarizing currents, a current-threshold relationship (indicating inward and outward rectification), and the recovery of excitability following supramaximal activation. The method was tested on 50 healthy volunteers, stimulating the median nerve at the wrist and recording the antidromic compound sensory nerve action potential (SNAP) from digit 2. The excitability measurements were similar, where comparisons were possible, with published sensory nerve data, and confirmed differences from motor nerves, particularly in strength-duration behavior and recovery cycle, likely to reflect functional differences between sensory and motor nerves. Although slower than for motor nerves, the sensory nerve recordings were sufficiently quick (16 to 18 min) to allow them to be included in routine clinical studies. We propose that this method, which provides quite different and complementary information about nerve function to conventional conduction studies, provides a useful new approach for exploring the pathophysiology of sensory neuropathies. PMID- 11410916 TI - Myostatin, insulin-like growth factor-1, and leukemia inhibitory factor mRNAs are upregulated in chronic human disuse muscle atrophy. AB - Human disuse muscle atrophy frequently accompanies orthopedic injury, arthritis, or bed rest, and recovery is often incomplete despite current rehabilitation programs. We have studied the vastus lateralis muscle in 12 patients with chronic disuse atrophy associated with chronic osteoarthritis of the hip both preoperatively and after total hip arthroplasty. Semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) demonstrated an increase in the level of expression of myostatin, insulin-like growth factor-1 (IGF-1) and leukemia inhibitory factor (LIF) mRNAs compared to healthy control muscle. In all patients there was a significant correlation preoperatively between increasing myostatin mRNA expression and reduction in type 2A and 2B fiber area. In the 8 female patients there was a significant correlation between increased myostatin mRNA expression and the atrophy factor calculated for 2A and 2B fiber types preoperatively. We hypothesize that a complex interaction occurs between muscle growth regulating factors in the genesis of muscle wasting. Our results indicate that myostatin is a muscle-wasting factor contributing to type 2B and 2A atrophy. Other muscle growth factors, such as IGF-1 and LIF, may be upregulated in a counterregulatory fashion or may be involved in the fiber type switching seen in disuse muscle wasting. PMID- 11410917 TI - A normative study on human facial F waves. AB - F waves from the nasalis muscle were obtained bilaterally after transcutaneous constant-current stimulation of the facial nerve in 37 of 42 (88%) healthy volunteers examined. F waves were of varying shape and latency. Standard parameters (latencies, amplitudes, ratios, chronodispersion) of F-wave analysis were assessed. Of these, minimum and mean F-wave latencies, the F ratio, and F wave and peripheral conduction times (PCT) were approximately normally distributed, with a low standard deviation. Latencies and conduction times exhibited a significantly positive correlation with body height, as has been demonstrated for the F-wave latencies in limb muscles. Side and sex differences were present but without statistical significance. Chronodispersion and chronodispersion range, F-wave amplitudes, and F-wave frequencies were broadly scattered. It is concluded that F-wave latencies and conduction times are best suited to define a range of reference values. Moreover, facial F waves bear characteristics similar to those described for F waves recorded from limb muscles. PMID- 11410918 TI - Effects of FK506 on nerve regeneration and reinnervation after graft or tube repair of long nerve gaps. AB - We compared the effects of FK506 administration on regeneration and reinnervation after sciatic nerve resection and repair with an autologous graft or with a silicone tube leaving a 6-mm gap in the mouse. Functional reinnervation was assessed by noninvasive methods to determine recovery of motor, sensory, and sweating functions in the hindpaw over 4 months after operation. Morphometric analysis of the regenerated nerves was performed at the end of follow-up. The nerve graft allowed for faster and higher levels of reinnervation in the four functions tested than silicone tube repair. Treatment with FK506 (for the first 9 weeks only) resulted in a slight, although not significant, improvement of the onset of reinnervation and of the maximal degree of recovery achieved after autografting. The recovery of pain sensibility and of the compound nerve action potentials in the digital nerves, which directly depend on axonal regeneration, showed better progression with FK506 than reinnervation of muscles and sweat glands, which require reestablishment of synaptic contacts with target cells. The myelinated fibers in the regenerated nerve showed a more mature appearance in the FK506-treated rats. However, FK506 showed a marginal effect in situations in which regeneration was limited, as in a silicone tube bridging a 6-mm gap in the mouse sciatic nerve. In conclusion, treatment with FK506 improved the rate of functional recovery after nerve resection and autograft repair. PMID- 11410919 TI - Nerve conduction changes in patients with mitochondrial diseases treated with dichloroacetate. AB - Serial measurements of nerve conduction velocities and amplitudes were performed in 27 patients with congenital lactic acidemia over 1 year of sodium dichloroacetate (DCA) administration. Patients were treated with oral thiamine (100 mg) and DCA (initial dose of 50 mg/kg) daily. Nerve conduction velocity and response amplitude were measured in the median, radial, tibial, and sural nerves at 0, 3, 6, and 12 months, and plasma DCA pharmacokinetics were measured at 3 and 12 months. Baseline electrophysiologic parameters in this population were generally below normal but as a group were within 2 standard deviations of normal means. Although symptoms of neuropathy were reported by only three patients or their families, nerve conduction declined in 12 patients with normal baseline studies, and worsening of nerve conduction occurred in the two who had abnormalities at baseline. Peripheral neuropathy appears to be a common side effect during chronic DCA treatment, even with coadministration of oral thiamine. Nerve conduction should be monitored during DCA treatment. PMID- 11410920 TI - Comparison of interpolation and central activation ratios as measures of muscle inactivation. AB - The objective of this study was to investigate different methods of estimating muscle inactivation, derived from single and multiple voluntary contractions. Ten subjects performed maximal and submaximal leg extensor contractions to determine an interpolation (IT) or central activation ratio (CAR). A superimposed evoked force was compared with the force output of either a voluntary (CAR) or resting evoked contraction (IT ratio), or the ratios were inserted into regression equations (linear, polynomial, exponential). Linear-regression estimates of CAR using doublets and tetanus provided physiologically inaccurate values. Whereas IT ratios using doublets (IT-doublet) and tetanus (IT-tetanus) had a significant difference in only one interaction, IT-tetanus and CAR using a tetanus (CAR tetanus) estimates provided the most extensive correlation within and between measures. Thus, tetanic stimulation superimposed upon single maximal or multiple contractions seems to provide the most valid measure of muscle inactivation when using the interpolated-twitch technique. PMID- 11410921 TI - Do nerve conduction studies predict the outcome of carpal tunnel decompression? AB - Patients (n = 3336) who had been tested for suspected carpal tunnel syndrome (CTS) were contacted by postal questionnaire and their opinions sought of the overall subjective outcome of any treatments using a five-point rating scale. Operative results were compared with preoperative nerve conduction, clinical and demographic variables, and a multiple logistic regression analysis used to identify significant prognostic factors. Among 1268 surgical procedures that were identified, the overall surgical success rate was 69%. Preoperative nerve conduction study findings, greater age, lower symptom scores, longer disease duration, and male gender were significant predictors of poor outcome. Nerve conduction studies had the strongest effect, with patients with middle-grade abnormalities having better results than those with either very severe or no abnormality. Surgical carpal tunnel decompression has a significant failure rate. The preoperative nerve conduction studies account for a proportion of the total variation in outcome. PMID- 11410922 TI - Stimulated single fiber electromyography in the mouse: techniques and normative data. AB - As the number of new transgenic mouse models of human neuromuscular disease continues to increase, the development of sophisticated electrophysiologic techniques for assessing the peripheral nervous system in these models has become important. Neuromuscular junction (NMJ) dysfunction, in particular, is often not detectable by morphologic or other techniques. To enable sensitive testing of murine NMJ function, we developed and tested a method for stimulated single fiber electromyography (S-SFEMG) in the gastrocnemius muscles of anesthetized mice. Jitter was assessed by measuring the mean consecutive latency difference (MCD) of single fiber responses to sciatic nerve stimulation at 2 HZ. Mean MCD values in normothermic mice were in the range of 6-8 micros for different strains, with no MCD values exceeding 25 micros. Reduced core temperature (to 29 degrees--30 degrees C) resulted in increased jitter, whereas intubation and mechanical ventilation of mice did not alter these values. Intraperitoneal and intravenous injection of vecuronium, however, resulted in progressively increased jitter followed by blocking in continuously monitored fibers. These observations validate the utility of S-SFEMG in mice as an index of NMJ function under a variety of physiologic conditions, and suggest that a high safety factor for neuromuscular transmission exists at mouse NMJs. PMID- 11410923 TI - Standardization of anal sphincter electromyography: utility of motor unit potential parameters. AB - Advanced electromyography systems offer quantitative analysis of a number of motor unit potential (MUP) parameters. However, only limited data are available on the diagnostic usefulness of these parameters. In the present study, we compared the sensitivities of MUP parameters in revealing "neuropathic" changes in the external anal sphincter (EAS) muscles in 56 patients examined 5-240 months after damage to the cauda equina or conus medullaris. Using multi-MUP analysis, 20 MUPs were obtained from patients' EAS muscles. Their MUP parameters were compared with normative data from 64 controls. The diagnostic sensitivities of mean values/"outliers" of MUP parameters for detecting neuropathic EAS muscles were calculated (area 25%/30%; number of turns 18%/29%; size index 13%/24%; thickness 18%/18%; amplitude 17%/17%; spike duration 20%/9%; duration 15%/12%; number of phases 15%/11%; and their combination 51%/52%). Altogether, the cumulative sensitivity of multi-MUP analysis using both mean values and "outliers" was 62%. The combination of MUP parameters improves the diagnostic yield of MUP analysis, but the influence on specificity remains unknown. PMID- 11410924 TI - Spontaneous motor unit behavior in human thenar muscles after spinal cord injury. AB - Our first aim was to characterize spontaneous motor unit activity in thenar muscles influenced by chronic cervical spinal cord injury. Thenar surface electromyography (EMG), intramuscular EMG, and abduction and flexion forces were recorded. Subjects were instructed to relax for 2 min. Units still firing after 10 s were considered spontaneously active. Two distinct patterns of spontaneous unit activity were recorded. Units either fired tonically at a mean frequency of 6.1 HZ or were active sporadically (2.2 HZ). Stimuli (e.g., light touch of nearby skin) were then used to influence tonic spontaneous unit activity. Most stimuli produced a change in firing frequency, usually a temporary increase, but sometimes unit frequency decreased or new activity was initiated. Inputs to these motoneurons clearly make important contributions to changes in unit activity. However, the difficulty that subjects had in stopping unit activity, and the initiation of activity when subjects relaxed, suggest that the source of spontaneity may be the motoneuron itself. PMID- 11410925 TI - Adynamic ileus in severe Guillain-Barre syndrome. AB - Adynamic ileus occurred in 17 of 114 patients (15%) with severe Guillain-Barre syndrome (GBS). Cardiovascular dysautonomia coincided with ileus in only five patients. In four patients, mechanical ventilation and immobilization could be implicated; and in eight patients, preexisting conditions such as prior abdominal surgery or incremental doses of opioids could be linked to ileus. Medical management was successful in all patients. Patients with GBS infrequently develop ileus as a manifestation of dysautonomia. PMID- 11410926 TI - Transcarpal motor conduction velocity in carpal tunnel syndrome. AB - Transcarpal motor conduction to abductor pollicis brevis (APB) was evaluated in 43 patients (70 hands) with suspected carpal tunnel syndrome (CTS). Transcarpal motor conduction was abnormal in 80% of hands compared with 11.5% with prolongated distal motor latency from wrist stimulation. Transcarpal motor conduction was comparable in sensitivity with transcarpal sensory conduction and 2nd lumbrical-interosseous latency difference. Transcarpal motor conduction is a sensitive test for diagnosis of CTS. Sensory fibers were no more susceptible than motor fibers to compression in the carpal tunnel, and fibers to APB were as susceptible as those to the 2nd lumbrical muscle. PMID- 11410927 TI - Focal myopathy mimicking posterior interosseous nerve syndrome. AB - A 25-year-old man developed weakness of extension of the right index, middle, and fourth fingers at the metacarpophalangeal joints, over 2 years. No sensory deficit was present. Nerve conduction studies, including the right radial nerve, were within normal limits. Needle electromyographic (EMG) examination showed myopathic changes that were limited to the right extensor digitorum communis and extensor indicis proprius muscles. An intravenous edrophonium chloride test had no effect on weakness and repetitive stimulation showed no significant decremental response. An EMG-guided open biopsy of the extensor digitorum communis muscle revealed severe myopathic changes. Evaluation for the cause of myopathic involvement was negative. After 13 months, clinical examination and electrophysiological studies showed no significant progression. This case exemplifies the fact that a focal myopathy may mimic an entrapment neuropathy. PMID- 11410928 TI - Critical illness myopathy. PMID- 11410932 TI - Arterial calcification: a review of mechanisms, animal models, and the prospects for therapy. AB - The causes of arterial calcification are beginning to be elucidated. Macrophages, mast cells, and smooth muscle cells are the primary cells implicated in this process. The roles of a variety of bone-related proteins including bone morphogenetic protein-2 (BMP-2), matrix Gla protein (MGP), osteoprotegerin (OPG), osteopontin, and osteonectin in regulating arterial calcification are reviewed. Animals lacking MGP, OPG, smad6, carbonic anhydrase isoenzyme II, fibrillin-1, and klotho gene product develop varying extents of arterial calcification. Hyperlipidemia, vitamin D, nicotine, and warfarin, alone or in various combinations, produce arterial calcification in animal models. MGP has recently been discovered to be an inhibitor of bone morphogenetic protein-2, the principal osteogenic growth factor. Many of the forces that induce arterial calcification may act by disrupting the essential post-translational modification of MGP, allowing BMP-2 to induce mineralization. MGP requires gamma-carboxylation before it is functional, and this process uses vitamin K as an essential cofactor. Vitamin K deficiency, drugs that act as vitamin K antagonists, and oxidant stress are forces that could prevent the formation of GLA residues on MGP. The potential role of arterial apoptosis in calcification is discussed. Potential therapeutic options to limit the rate of arterial calcification are summarized. PMID- 11410933 TI - Centchroman, a selective estrogen receptor modulator, as a contraceptive and for the management of hormone-related clinical disorders. AB - DL-Centchroman (67/20; INN: Ormeloxifene) synthesized at the Central Drug Research Institute, Lucknow, is a nonsteroidal once-a-week oral contraceptive. It was introduced in Delhi in July, 1991, marketed in India in 1992 as Saheli and Choice-7 (Hindustan Latex Ltd., Thiruvananthapuram) and Centron (Torrent Pharmaceuticals India Ltd., Ahmedabad), and included in the National Family Welfare Programme in 1995.5 According to post-marketing surveillance, approximately 100,000 women were using this pill and approximately 1100,000 menstrual cycles were covered until 1996. It is a unique need-oriented contraceptive being effective when taken immediately after coitus or routinely as a weekly pill and has the advantage of less frequent administration. Its contraceptive action is quickly reversible. It has long terminal serum halflife of 168 hr in women and exhibits duration of anti-implantation/estrogen antagonistic action of 120 hr, despite a short (24.1 hr) serum halflife, in the rat. In lactating women, it is excreted in milk in quantities considered unlikely to cause any deleterious effect on suckling babies. In phase II and III multicentric trials as a contraceptive, children born of method-and-user failure pregnancies showed normal milestones, without any congenital anomaly. Reports of its promising action in the management of certain hormone-related clinical disorders are available. It has an excellent therapeutic index and is considered safe for chronic administration. PMID- 11410934 TI - The active site of HIV-1 protease. AB - The active site of the homodimeric HIV-1 protease includes six amino acids (triads AspThrGly found in each monomer) in amino acid positions 25 to 27 and 25' to 27'. Up to now, the role of Thr26 and Thr26', and Gly27 and Gly27', is unknown. It is hypothesized that strong hydrogen-bonding forces between the Thr26 and Thr26' residues stabilize the conformational state of the active site, and that the function of Gly27 and Gly27' is to accommodate and bind a substrate in a position in which the catalytic Asp25 and Asp25' carboxylate groups can attack the amide moiety of a substrate. PMID- 11410931 TI - The ubiquitin-proteasome pathway and proteasome inhibitors. AB - The ubiquitin-proteasome pathway has emerged as a central player in the regulation of several diverse cellular processes. Here, we describe the important components of this complex biochemical machinery as well as several important cellular substrates targeted by this pathway and examples of human diseases resulting from defects in various components of the ubiquitin-proteasome pathway. In addition, this review covers the chemistry of synthetic and natural proteasome inhibitors, emphasizing their mode of actions toward the 20S proteasome. Given the importance of proteasome-mediated protein degradation in various intracellular processes, inhibitors of this pathway will continue to serve as both molecular probes of major cellular networks as well as potential therapeutic agents for various human diseases. PMID- 11410936 TI - Introduction to the Rhizocephala (Crustacea: Cirripedia). AB - Knowledge about the Rhizocephala (parasitic barnacles) has increased exponentially over the past two decades. This introduction broadly reviews this progress, touching on rhizocephalan morphology, life-cycles, larval biology, and the effects on the crustacean hosts. Members representing both orders, the Kentrogonida and Akentrogonida, are discussed. The recent discovery regarding the vermigon stage further highlights the intricacy of the kentrogonid life-cycle. Because rhizocephalans are found in most world regions, occurring on their respective hosts from the deep ocean to freshwater, their importance is now being recognized. PMID- 11410937 TI - Comparative ultrastructure of the root system in rhizocephalan barnacles (Crustacea: Cirripedia: Rhizocephala). AB - Rhizocephalan barnacles are parasites of Crustacea. They lack even the rudiments of an alimentary canal, but infiltrate their hosts with a nutrient-absorbing system of rootlets. We review the ultrastructure of the rootlets using light microscopy, SEM, and TEM in nine species from five families, representing both suborders of the Rhizocephala: from the Kentrogonida Peltogaster paguri, P. curvatus, Peltogasterella sulcata, Cyphosaccus norvegicus (Peltogastridae); Lernaeodiscus porcellanae (Lernaeodiscidae); and Sacculina carcini (Sacculinidae); and from the Akentrogonida Clistosaccus paguri (Clistosaccidae); Chthamalophilus delagei, and Boschmaella japonica (Chthamalophilidae). With the exception of Chthamalophilus delagei, the root system of the investigated species shares numerous apomorphies at the ultrastructural level and displays at all levels specializations that maximize the surface area. The rootlets consist of a cuticle, an epidermis and a subjacent layer of axial cells that often, but not always surround, a central lumen. The rootlets are at all times enclosed in a less than 0.5 microm thick cuticle, which is never molted. The cuticle consists of an inner homogeneous layer with a slightly fibrous structure and an outer, less than 15-nm thick electron-dense layer, from which numerous microcuticular projections extend into the hemolymphatic space of the host. The microcuticular projections consist of the outer electron-dense layer and sometimes a core of the more translucent homogeneous layer. They vary among the species from being simple in Sacculina carcini to exhibiting complex branching patterns in Peltogasterella sulcata and Cyphosaccus norvegicus. Beneath the cuticle the epidermal plasma membrane is thrown into irregularly shaped projections. The epidermal cells are joined by long septate junctions and exhibit the characteristics of a transporting epithelium. Experiments with acid phosphatase revealed activity both in the epidermis and among the microcuticular projections. The projections may therefore form a domain that is important in absorption and extracellular digestion of nutrients from the host. The axial cells contain abundant endoplasmic reticulum and seem to convert absorbed carbohydrates into lipid, which is stored in large droplets. Subepidermal muscle cells cause sinuous movements of the rootlets, but it remains unknown how nutrients are transported along the rootlets towards the external reproductive body. In C. delagei the single, bladder-shaped rootlet lacks both the apical projections in the epidermis, the electron-dense cuticle layer, and the microcuticular projections. We review previous studies on the rhizocephalan root system and discuss functional and phylogenetic aspects of the morphology. PMID- 11410938 TI - Cypris metamorphosis, injection and earliest internal development of theRrizocephalan Loxothylacus panopaei (Gissler). Crustacea: Cirripedia: Rhizocephala: Sacculinidae. AB - Rhizocephala is a group of crustaceans that exclusively parasitizes other crustaceans. It is taxonomically placed within the class Cirripedia, the barnacles, with which it shares a unique larval type, the cyprid. The main objective of the cyprid is to find and irreversibly attach to a suitable substratum and initiate metamorphosis. In the presumed sister group to Rhizocephala, the true barnacles or Thoracica, metamorphosis leads to a juvenile filter-feeding version of the adult organism. In Rhizocephala the female cyprid settles on the integument of a crustacean and undergoes metamorphosis into a kentrogon that possesses a hollow cuticular-tube structure, the stylet, which penetrates the integument of the host and acts as a guide tube for the prospective internal parasite. The first, hitherto unknown endoparasitic stage of a rhizocephalan, the vermigon, was recently discovered (Glenner and Hoeg [1995] Nature 377:147-150) and its migration through the hemolymph of the host, as well as its internal development, was described in Glenner et al. ([2000] Mar Biol 136:249-257). The present article provides detailed information on kentrogon and vermigon formation, the injection process, and the succeeding developmental stages up to the stage of the earliest primordium reported from the literature. The anlage of the ovary is traced back to the free-swimming cypris stage and it is implied that the mesoderm and ectoderm of the endoparasite are already differentiated in the cyprid. PMID- 11410939 TI - NMR spectroscopy quantitation. PMID- 11410940 TI - Common processing of in vivo MR spectra. AB - This introductory article addresses approaches currently in use to process in vivo spectra. First, a brief overview is given of the information content represented by the parameters of MR signals. Subsequently, common steps in the processing of MR spectra such as pre-processing, normalisation and quantification and the use of prior knowledge are described. Finally, some prospects for more advanced processing are given. PMID- 11410941 TI - MR spectroscopy quantitation: a review of time-domain methods. AB - In this article an overview of time-domain quantitation methods is given. Advantages of processing the data in the measurement domain are discussed. The basic underlying principles of the methods are outlined and from them the situations under which these algorithms perform well are derived. Also an overview of methods to preprocess the data is given. In that respect, signal-to noise and/or resolution enhancement, the removal of unwanted components and corrections for model imperfections are discussed. PMID- 11410942 TI - MR spectroscopy quantitation: a review of frequency domain methods. AB - There has been a vast increase in applications of magnetic resonance spectroscopy (MRS) in biomedical research during the last few years. This is not surprising since MRS provides both in vivo and in vitro a non-invasive tool for various biochemical and biomedical studies. There are also expectations that clinical MRS will have an important role as a diagnostic tool. An essential prerequisite for the future success of MRS for applicability in biomedical sciences will be accurate and biochemically relevant data analysis (at as high a level of automation as possible). This review briefly describes principles of the methodology available for advanced quantitative data analysis in the frequency domain. Various biomedical applications are discussed in order to illustrate the practical aspects of the analyses and to show the applicability and power of biochemical prior knowledge-based lineshape fitting analysis. PMID- 11410943 TI - Automatic quantitation of localized in vivo 1H spectra with LCModel. AB - The LCModel method analyzes an in vivo spectrum as a Linear Combination of Model in vitro spectra from individual metabolite solutions. Complete model spectra, rather than individual resonances, are used in order to incorporate maximum prior information into the analysis. A nearly model-free constrained regularization method automatically accounts for the baseline and lineshape in vivo without imposing a restrictive parameterized form on them. LCModel is automatic (non interactive) with no subjective input. Approximately maximum-likelihood estimates of the metabolite concentrations and their uncertainties (Cramer-Rao lower bounds) are obtained. LCModel analyses of spectra from users with fields from 1.5 to 9.4 T and a wide range of sequences, particularly with short TE, are used here to illustrate the capabilities and limitations of LCModel and proton MRS. PMID- 11410944 TI - Wavelets and related time-frequency techniques in magnetic resonance spectroscopy. AB - We survey the various applications in MRS of the wavelet transform and related time-frequency methods. For the sake of completeness, we first quickly review the mathematical tools needed. PMID- 11410945 TI - NMR spectral quantitation by principal component analysis. AB - The use of principal component analysis (PCA) for simultaneous spectral quantitation of a single resonant peak across a series of spectra has gained popularity among the NMR community. The approach is fast, requires no assumptions regarding the peak lineshape and provides quantitation even for peaks with very low signal-to-noise ratio. PCA produces estimates of all peak parameters: area, frequency, phase and linewidth. If desired, these estimates can be used to correct the original data so that the peak in all spectra has the same lineshape. This ability makes PCA useful not only for direct peak quantitation, but also for processing spectral data prior to application of pattern recognition/classification techniques. This article briefly reviews the theoretical basis of PCA for spectral quantitation, addresses issues of data processing prior to PCA, describes suitable and unsuitable datasets for PCA applications and summarizes the developments and the limitations of the method. PMID- 11410946 TI - Cramer-Rao bounds: an evaluation tool for quantitation. AB - The Cramer-Rao lower bounds (CRBs) are the lowest possible standard deviations of all unbiased model parameter estimates obtained from the data. Consequently they give insight into the potential performance of quantitation estimators. Using analytical CRB expressions for spectral parameters of singlets and doublets in noise, one is able to judge the precision as a function of spectral and experimental parameters. We point out the usefulness of these expressions for experimental design. The influence of constraints (chemical prior knowledge) on spectral parameters of the peaks of doublets is demonstrated and the inherent benefits for quantitation are shown. Abbreviations used: CRB Cramer-Rao lower bounds PMID- 11410947 TI - Current awareness. AB - In order to keep subscribers up-to-date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of NMR in biomedicine. Each bibliography is divided into 9 sections: 1 Books, Reviews ' Symposia; 2 General; 3 Technology; 4 Brain and Nerves; 5 Neuropathology; 6 Cancer; 7 Cardiac, Vascular and Respiratory Systems; 8 Liver, Kidney and Other Organs; 9 Muscle and Orthopaedic. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. PMID- 11410948 TI - A 4D approach to the analysis of functional brain images: application to FMRI data. AB - This paper presents a new approach to functional magnetic resonance imaging (FMRI) data analysis. The main difference lies in the view of what comprises an observation. Here we treat the data from one scanning session (comprising t volumes, say) as one observation. This is contrary to the conventional way of looking at the data where each session is treated as t different observations. Thus instead of viewing the v voxels comprising the 3D volume of the brain as the variables, we suggest the usage of the vt hypervoxels comprising the 4D volume of the brain-over-session as the variables. A linear model is fitted to the 4D volumes originating from different sessions. Parameter estimation and hypothesis testing in this model can be performed with standard techniques. The hypothesis testing generates 4D statistical images (SIs) to which any relevant test statistic can be applied. In this paper we describe two test statistics, one voxel based and one cluster based, that can be used to test a range of hypotheses. There are several benefits in treating the data from each session as one observation, two of which are: (i) the temporal characteristics of the signal can be investigated without an explicit model for the blood oxygenation level dependent (BOLD) contrast response function, and (ii) the observations (sessions) can be assumed to be independent and hence inference on the 4D SI can be made by nonparametric or Monte Carlo methods. The suggested 4D approach is applied to FMRI data and is shown to accurately detect the expected signal. PMID- 11410949 TI - A role for left temporal pole in the retrieval of words for unique entities. AB - Both lesion and functional imaging studies have implicated sectors of high-order association cortices of the left temporal lobe in the retrieval of words for objects belonging to varied conceptual categories. In particular, the cortices located in the left temporal pole have been associated with naming unique persons from faces. Because this neuroanatomical-behavioral association might be related to either the specificity of the task (retrieving a name at unique level) or to the possible preferential processing of faces by anterior temporal cortices, we performed a PET imaging experiment to test the hypothesis that the effect is related to the specificity of the word retrieval task. Normal subjects were asked to name at unique level entities from two conceptual categories: famous landmarks and famous faces. In support of the hypothesis, naming entities in both categories was associated with increases in activity in the left temporal pole. No main effect of category (faces vs. landmarks/buildings) or interaction of task and category was found in the left temporal pole. Retrieving names for unique persons and for names for unique landmarks activate the same brain region. These findings are consistent with the notion that activity in the left temporal pole is linked to the level of specificity of word retrieval rather than the conceptual class to which the stimulus belongs. PMID- 11410950 TI - Human cortical processing of colour and pattern. AB - The present study investigates human visual processing of simple two-colour patterns using a delayed match to sample paradigm with positron emission tomography (PET). This study is unique in that we specifically designed the visual stimuli to be the same for both pattern and colour recognition with all patterns being abstract shapes not easily verbally coded composed of two-colour combinations. We did this to explore those brain regions required for both colour and pattern processing and to separate those areas of activation required for one or the other. We found that both tasks activated similar occipital regions, the major difference being more extensive activation in pattern recognition. A right sided network that involved the inferior parietal lobule, the head of the caudate nucleus, and the pulvinar nucleus of the thalamus was common to both paradigms. Pattern recognition also activated the left temporal pole and right lateral orbital gyrus, whereas colour recognition activated the left fusiform gyrus and several right frontal regions. PMID- 11410951 TI - Early discrimination of coherent versus incoherent motion by multiunit and synaptic activity in human putative MT+. AB - A laminar probe was chronically implanted in human putative MT+. The area was specifically responsive to globally coherent visual motion, a crucial aspect of the perception of movement through space. The probe contained 23 microcontacts spaced every 175 microm in a linear array roughly perpendicular to the cortical surface. Current-source density (CSD) and multiunit activity (MUA) were recorded while viewing initially stationary random dot patterns that either moved incoherently or dilated from the central fixation. Onset of visual motion evoked large MUA/CSD activity, with coherent motion evoking earlier and faster-rising MUA/CSD activity than incoherent, in both superficial and deep pyramidal layers. The selective response, peaking at approximately 115 ms, was especially large in deep pyramids, providing evidence that information necessary for visual flow calculations is projected from MT+ at an early latency to distant structures. The early onset of differential MUA/CSD implies that the selectivity of this area does not depend on recurrent inhibition or other intrinsic circuitry to detect coherent motion. The initially greater increase of MUA to coherent stimuli was followed by a greater decrease beginning at approximately 133 ms, apparently because of recurrent inhibition. This resulted in the total MUA being greater to incoherent than coherent stimuli, whereas total rectified CSD was overall greater to coherent than to incoherent stimuli. However, MUA distinguished stationary from moving stimuli more strongly than did CSD. Thus, while estimates of total cell firing (MUA), and of total synaptic activity (CSD) generally correspond to previously reported BOLD results, they may differ in important details. PMID- 11410952 TI - fMRI investigation of sentence comprehension by eye and by ear: modality fingerprints on cognitive processes. AB - The neural substrate underlying reading vs. listening comprehension of sentences was compared using fMRI. One way in which this issue was addressed was by comparing the patterns of activation particularly in cortical association areas that classically are implicated in language processing. The precise locations of the activation differed between the two modalities. In the left inferior frontal gyrus (Broca's area), the activation associated with listening was more anterior and inferior than the activation associated with reading, suggesting more semantic processing during listening comprehension. In the left posterior superior and middle temporal region (roughly, Wernicke's area), the activation for listening was closer to primary auditory cortex (more anterior and somewhat more lateral) than the activation for reading. In several regions, the activation was much more left lateralized for reading than for listening. In addition to differences in the location of the activation, there were also differences in the total amount of activation in the two modalities in several regions. A second way in which the modality comparison was addressed was by examining how the neural systems responded to comprehension workload in the two modalities by systematically varying the structural complexity of the sentences to be processed. Here, the distribution of the workload increase associated with the processing of additional structural complexity was very similar across the two input modalities. The results suggest a number of subtle differences in the cognitive processing underlying listening vs. reading comprehension. PMID- 11410953 TI - Putting your health before the cart. PMID- 11410954 TI - ACE inhibitor inhibits atherosclerosis. PMID- 11410956 TI - By the Way, Doctor: Statins and Fracture Risk. PMID- 11410955 TI - Ask the doctor. I take medicine to lower my cholesterol. My doctor says that I've reached my target (my LDL is just under 100 mg/dL), but I wonder whether I should try to get my cholesterol even lower. I haven't heard that you'll get hurt by taking too much of these drugs, so why shouldn't I increase my dose? PMID- 11410958 TI - By the Way, Doctor: What's the Story with PPA? PMID- 11410957 TI - By the Way, Doctor: Diet Pills and Heart-Valve Disease. PMID- 11410959 TI - Navigating the Medicaid delivery system. PMID- 11410960 TI - PDA establishes position statement on cola contracts in schools. AB - The soft drink industry promises that it will do everything possible to persuade even more Americans to drink even more soda pop more often. Parents and health care providers need to recognize soft drinks for what they are--liquid candy--and do everything possible to return these beverages to their former, reasonable role as an occasional treat. School districts should not auction themselves off to the highest bidder for exclusive pouring rights contracts with soft drink companies. These deals profit the soft drink companies and, to a much lesser degree, the school districts at the expense of the students' health. PMID- 11410961 TI - Gold foils. PMID- 11410962 TI - "First wave" results of PDA's marketing program show measurable achievements. PMID- 11410963 TI - The rape of a colleague. PMID- 11410964 TI - Periodontal diseases and systemic diseases: recognition & liability. PMID- 11410965 TI - High hopes for the dental summit. PMID- 11410966 TI - Improved access to dental care for residents of Chester County. PMID- 11410968 TI - A challenge. PMID- 11410969 TI - Clinical evaluation of "veraviewpocs" digital panoramic X-ray system. PMID- 11410970 TI - Preparations for Cerec 3: where are the limits? PMID- 11410971 TI - Correlation with Cerec scan-experience with Cerec 3 in a large practice with an integrated dental laboratory. PMID- 11410972 TI - Residual dentin thickness after 1.2-mm shoulder preparation for Cerec crowns. AB - Since the introduction of crown software for Cerec, it has been possible to produce full-ceramic crown restorations. Despite the excellent material quality of industrially made ceramic blocks and the good clinical long-term success of these ceramics in inlay- and onlay preparations, extended preparation with high loss of dental tissue is incident to crown restorations, which may endanger the vitality of the respective tooth. The aim of this experimental study was to determine the residual dentin thickness after preparation of an extended circular shoulder in the molar and premolar region. On extracted human molars and premolars, 1.2-mm-wide circular shoulders with rounded inner edges were prepared, and the thickness of the residual dentin wall was measured under a microscope. Using this preparation method, it was found that only the group of maxillary molars showed an average residual dentin thickness of more than 0.7 mm. PMID- 11410973 TI - Infection control procedures used in conjunction with computed dental radiography. AB - Infection control guidelines for dental radiography have been modified since 1986, when the American Dental Association and the Centers for Disease Control and Prevention supported the concept of "universal blood and body fluid precautions." With the introduction of computed digital radiography, hardware manufacturers recommend that alternative infection control techniques are necessary to prevent potential damage to the digital x-ray sensors placed inside the patient's mouth. Thirty first-year dental hygiene students were asked to insert and remove a Schick CDR number 2 size intraoral digital x-ray sensor into modified Rinn XCP bitewing bite blocks and a modified Rinn Snap-a-ray five times with each of the recommended infection control covers. Reduced rates of cross contamination are possible if the plastic barrier envelope has an additional latex finger cot stretched over it and the x-ray sensor. Sole usage of a latex finger cot will result in a reduced incidence of contamination, but still not to acceptable levels. However, a plastic barrier envelope placed over the x-ray sensor and over the modified XCP bite block together or a covered sensor in a Snap-a-ray under normal conditions does not result in a perforation and is least likely to result in cross contamination. PMID- 11410974 TI - The optical conditioning of Cerec preparations with scan spray. AB - Coating the surfaces with an anti-reflection substance before taking the optical impression is indispensable for exposures with the Cerec camera. Up to now, this has been done mainly by powdering with titanium dioxide. Errors which have a negative influence on the result can, of course, arise during the coating process. Although powdering is a proven method which has led to outstanding results in many thousands of restorations, it represents a weak spot in the clinical procedure. In the present study, the handling and data quality of an innovative spray for applying an anti-reflection coat to restoration surfaces were examined. The values obtained were chiefly displayed in the Section window of the Cerec 3 software. Despite a few small problems, handling can be considered good and safe. The primary advantage consists in the display of the preparation boundaries, since the coated surface can be dried with an air stream. With regard to detail reproduction, data quality, and coat thickness, the quality of the optical impression has reached a new level. The present study shows that considerable errors can arise in the restoration from powdering, due partly to material buildup. The Scan Spray tested can be recommended for clinical use. PMID- 11410975 TI - Correlation with Cerec 3 for end teeth: single-side correlation. AB - Correlation is not necessarily a problem for end teeth any longer. All the advantages of Correlation, anatomical occlusal surfaces, and very slight vertical occlusion adjustment are also available with the described method for end teeth and gaps. PMID- 11410976 TI - The fallacy of goal fulfillment. PMID- 11410978 TI - Documenting the starting point: results of PDA's baseline consumer awareness survey. PMID- 11410979 TI - Lessons from medicine's miscalculations. PMID- 11410980 TI - Dentistry today: putting profit before patients. PMID- 11410981 TI - Clinical judgement and decision making in endodontics. AB - Clinical judgement in endodontics consists of much more than diagnosis and treatment planning for the affected tooth. The issues involved in clinical judgement and decision making can be summarised by three questions: 1. Is endodontic treatment appropriate for the patient? Endodontic treatment should be undertaken only as part of an agreed, comprehensive treatment plan that takes into account patient concerns as well as objective clinical findings. 2. How difficult is the endodontic treatment? The difficulty of the case should be balanced with the skill and experience of the dentist, in deciding whether to manage the case in general practice or to refer the patient to an endodontist. The use of a standard form for assessing the difficulty of each endodontic case will aid in consistent, systematic assessment of patients. An example of such a form is provided. 3. What is the prognosis for the tooth? The outcome of endodontic treatment depends not only on the endodontic treatment but on other factors such as restorability and periodontal status. The prognosis will be compromised by procedural problems and by restorative and periodontal factors. In all but routine cases, the steps involved in decision making may be more complex and less easily resolved than the practical clinical aspects of endodontic therapy. PMID- 11410982 TI - Apexification following gross overfilling of gutta-percha. PMID- 11410983 TI - The consequences of internal tooth bleaching. PMID- 11410984 TI - Endodontic management of internal inflammatory resorption--report of a case. PMID- 11410985 TI - Successful apexification in the presence of a periapical cyst--a case report. PMID- 11410986 TI - Mandibular block injections--the inner game. AB - This paper is a sequel to a previous article published in the April 1998 issue of this journal (1). Whereas the earlier work concentrated largely on the anatomy and geometry of the Gow-Gates block, the present paper is more directed towards the mental barriers encountered when a clinician is confronted by a whole new set of conceptual demands. Paradoxically, the concepts underlying the Gow-Gates technique are almost diametrically opposite to what has traditionally been taught in dental schools in relation to the inferior alveolar nerve block. The author's central hypothesis is that the explanation for indifferent results experienced by dentists in their first attempts at the Gow-Gates technique lies primarily in mindsets acquired from experience with the inferior alveolar block. It is suggested that the subconscious mind can be as powerful a determinant of human behaviour as the conscious mind, a position more in line with the findings of Freud and Jung than with the cognitivist vogue enthusiastically embraced by contemporary academic psychologists. It is further postulated that the subconscious is amenable to conscious guidance, a skill that can be enhanced by practice. PMID- 11410987 TI - A comparison of hydroxyl ion diffusion through root dentine from various calcium hydroxide preparations. PMID- 11410988 TI - Endodontics in Vietnam. PMID- 11410989 TI - Interview with an insider: how to pursue a successful old age. Interview by Judith McFadden. PMID- 11410990 TI - Customer for life: marketing oral health care to older adults. AB - Respect for and awareness of the needs of older patients from dental office staff will help such patients feel welcome in a practice. Marketing to older patients is built upon this foundation. In addition, there are other strategies for internal and external marketing aimed at older people. This article addresses the concept of turning aging patients into "customers for life." PMID- 11410991 TI - Missing the diagnosis of oral cancer: recognition & liability. PMID- 11410992 TI - Geezerhood. PMID- 11410993 TI - If your patients ask about dental unit waterlines. PMID- 11410994 TI - Morphological characteristics and distribution of the autonomic and sensitive innervation of the prostate in some animal species. AB - The sensitive and autonomic innervation of the prostate was studied in some mammals (rat, rabbit, horse, donkey and bull) using gold chloride impregnation. In all the species considered the gland was supplied with conspicuous innervation located in the thickness of the capsule and in trabeculae. The autonomic innervation was constituted by ganglion cells of different sizes (small, medium and large), isolated or in groups of large ganglia also, always located along the course of nerve bundles. The sensitive nervous component, placed more frequently in the most superficial layer of the capsule, consisted of delicate networks originating by more fibres and, above all, of capsulated receptors such as Pacini and Golgi-Mazzoni corpuscles. A characteristic finding, but only present in the horse, donkey and bull, was the helicoidal disposition of thin collagen fibres around thin nervous bundles that ended up to the point where the nerve trunk divided. PMID- 11410995 TI - Characterization of human bone cells in culture. AB - Osteoblast-like cells isolated from human bone bioptic specimens were established in culture. Their osteoblast-like phenotype was studied by biochemical, histochemical and immunohistochemical methods and by electron microscopy examination. Third-passage cell cultures exhibited high level of alkaline phosphatase activity and the exposure to human parathyroid hormone produced an increase of intracellular cAMP. Cultured cells were immunoreactive for type I and type III collagen, osteonectin, and fibronectin; when ascorbic acid and beta glycerophosphate were added, they synthesized a rich extracellular matrix. This characterization ensures the reliability of osteoblast-like cultures when they are used as experimental models. PMID- 11410996 TI - Morphometric dimensions of the mouse parotid glands of both sexes. AB - The goal of this research was to evaluate the morphometric dimensions of the different structures of male and female albino mouse parotid glands. The following morphometric dimensions were evaluated for the acini, intercalated ducts, striated ducts, excretory ducts and stroma: volume density, total compartmental volume, surface density, total external surface, surface-to-volume ratio, cell volume and absolute number of cells. Analysis of the results showed that the parotid gland mass was 43.7% greater (P < 0.01) in the male mice than in the females. This difference was due to the fact that the compartmental volumes of the acini, intercalated ducts and striated ducts were markedly higher in the male mice, 57.6% (P < 0.01), 253.1% (P < 0.01) and 91.1% (P < 0.05), respectively. The higher volume of the acinar morphological compartment was due to the total number of cells and average cell volume being higher in the male mice, 24.8% P < 0.01) and 47.7% (P < 0.01), respectively. Based on the results obtained, it was concluded that there are morphological differences between male and female parotid glands. These differences are detectable through morphometry, mainly in the morphological acinar and intercalated ducts compartments, which are more developed in male mice. PMID- 11410997 TI - A method for separation of heparin species from biological samples by ethanol precipitation of compounds solubilized in guanidine hydrochloride. AB - In this paper we describe a procedure to determine glycosaminoglycan and oligosaccharide composition of biological samples such as cell cultures or tissue explants. We demonstrate that heparin species of different molecular mass can be easily fractionated by sequential ethanol precipitation in 4.0 M guanidine hydrochloride. We studied by gradient polyacrylamide gel electrophoresis fractionation of standard heparin and heparin-derived oligosaccharides by anion exchange chromatography on DEAE-Sephacel resin eluted by increasing concentration of guanidine hydrochloride. The use of guanidine salts followed by sequential precipitation by increasing ethanol concentration allowed recovery of heparin and heparin-derived oligosaccharides. PMID- 11410998 TI - Langerhans cells differentiation: a three-act play. AB - Langerhans cells are the dendritic cells typical of stratified squamous epithelia and enjoy an intermediate state of differentiation as dendritic cells, since they are very efficient in uptaking and processing antigens but poor in presenting them to lymphocytes. These cells, and other dendritic cells as well, differentiate from bone marrow precursors, with circulating monocytes as intermediates. Langerhans cells leave the epidermis upon loading with antigen and further differentiate into mature dendritic cells while moving to peripheral lymphoid organs, where they elicit primary immune responses. The signals regulating differentiation of precursors into Langerhans cells and of this latter cells into mature dendritic cells have been unraveled thanks to in vitro studies and include a wide range of cytokines, some of which can actively inhibit this process. Also pro-B cells and thymic precursors can give raise in vitro to mature dendritic cells, but apparently not to Langerhans cells. Keratinocytes, the principal cell type of epidermis and mucosal stratified squamous epithelia, can provide for a wide range of cytokines influencing the differentiation of cells of dendritic lineage, in particular Langerhans cells. These cytokines include GM-CSF and TNF-alpha, that may be relevant also for the maintenance of the differentiated state of Langerhans cells within epidermis. Cell to cell contacts mediated by E-cadherin may also contribute signals from keratinocytes to differentiating Langerhans cells. The epidermis and mucosal squamous epithelia host other cell types, besides keratinocytes, and nerve fibers, that all can contribute signals to differentiating Langerhans cells. The differentiation of Langerhans cells within the epithelial microenvironment of skin and mucosae most probably results from the interaction of several players, that play different roles on this scene in different physiological and pathophysiological conditions. PMID- 11410999 TI - The anatomical resources on the Internet. AB - The main anatomical resources found on the Internet are listed and discussed here, concerning the topics of gross anatomy, neuroanatomy, microscopic anatomy and anatomical museums. The search is updated to September 2000, and all the listed Web sites can be used to build a multimedial anatomical lab. PMID- 11411000 TI - Lower limb amputation. Part 2: Rehabilitation--a 10 year literature review. AB - Ten years after the ISPO consensus conference on amputation surgery, a search of relevant publications in the Rehabilitation-prosthetics-literature over the years 1990-2000 was performed. The main key-words in this research were: "lower limb, amputation, human and rehabilitation". One hundred and four (104) articles were assessed by reading and from these the authors selected 24 articles. These articles are summarised, under several subheadings in this review article, focussing especially on quality of life, functional outcome and predictive factors. PMID- 11411001 TI - Lower limb amputation. Part 3: Prosthetics--a 10 year literature review. AB - This paper is intended as a follow-up to the ISPO Consensus Conference on Amputation Surgery. It reviews all the literature on lower limb prosthetics published after 1990. The review was considered under six categories: feet, knees, hips, thermoplastics, liners/suspension and computers. PMID- 11411002 TI - Major lower limb amputation following failed infrainguinal vascular bypass surgery: a prospective study on amputation levels and stump complications. AB - The effect of failed vascular bypass surgery on final amputation level and stump complications is the subject of debate. The aim of this prospective cohort study was to assess the influence of previous infrainguinal bypass surgery on amputees in the authors' centre. Over a three-year period, 234 amputations (219 patients) were performed for critical ischemia. The cause of ischemia was either peripheral obstructive arterial disease (POAD) or diabetes mellitus (DM). Forty-eight percent (48%) (113 amputations) had ipsilateral vascular bypass surgery prior to amputation and 52% (121 amputations) had not. Final amputation level and the post operative complications of infection, significant stump pain and delayed wound healing were used as the outcome measures for this study. At the end of the study period these outcome measures were used to compare the influence of previous bypass surgery on the two groups of amputees. There was a significantly higher rate of transfemoral amputations (TFA) (32.7%) vs. 16.5%; p < 0.05) and stump infection rate (42% vs. 23%; p < 0.05) in the bypass group. Significant stump pain (p = 0.23) and delayed wound healing (p = 0.24) was more prevalent in the bypass group although statistical significance could not be demonstrated. PMID- 11411003 TI - Standing interface stresses as a predictor of walking interface stresses in the trans-tibial prosthesis. AB - Interface pressures and shear stresses within the socket, in standing and walking, were measured for two unilateral, male, trans-tibial amputee subjects, during two sessions each. The ratios of equal weight-bearing standing stresses to peak walking stresses showed regional variation, ranging from 0.24:1 for pressure over the anterior region to 1.01:1 for resultant interface shear stress over the lateral region. Interface stresses in standing were only moderate predictors of peak walking stresses. The best correlation coefficient between standing in full weight-bearing and peak walking stress was 0.88 for pressure over the lateral region. As the amputees progressed from minimal to full weight-bearing in standing, and then to walking, the interface stresses increased in a nonlinear fashion, consistent with the assumption that the anterior tibia provides much resistance to the bending moment in the sagittal plane during walking. PMID- 11411004 TI - Polyurethane gel liner usage in the Oxford Prosthetic Service. AB - The objective was to investigate which lower limb amputees are using Alpha polyurethane gel liners and the effects of these on comfort and suspension of their prosthesis. A retrospective study was carried out of case records of all patients issued with Alpha cushion and locking liners between 1997 and the end of January 1999. The type of liner used was compared with age, sex, level and cause of amputation, time since amputation, comfort and suspension. Modified Stanmore/Harold-Wood mobility grades; duration of use and number of liners issued per patient were recorded. Sixteen (16) patients were identified who had been prescribed Alpha cushion liners. Improved comfort was reported by all. Forty (40) patients were identified who had been prescribed Alpha locking liners. Twenty (20) of these reported improved comfort and 10 improved suspension. The average time since amputation was 18.5 years for those using cushion liners and 14.1 years for locking liner users. Fifty-two (52) of all 56 patients using Alpha cushion and locking liners had mobility grades of 4 or more. Trauma was the most common cause of amputation. This group is a relatively mobile group of amputees. All those using cushion liners reported improved comfort. Some of the locking liner users reported improved comfort and suspension but this was not universally the case. PMID- 11411005 TI - A reciprocal walking orthosis hip joint for young paediatric patients with a variety of pathological conditions. AB - A growing trend in the use of reciprocal walking orthoses for infant paraplegic patients, and their application for control of the lower limbs in very young total body involved cerebral palsy patients, has created a need for smaller components. A prototype design of a hip joint has been produced which provides the following features: adjustable range of flexion/extension control; override on stops to permit sitting; high lateral rigidity; no lateral bearing play; very high rigidity in the sagittal plane; low friction bearings; high resistance to torque about the vertical axis. In addition a size envelope which is more in keeping with the dimensions of infant patients was an important objective. Comparisons were made of the computed structural properties of the prototype joint and existing routinely available standard orthotic hip joints. In each plane of loading the prototype joint had the highest identified structural property. The hinge-bearing material was tested in a representative joint with 200,000 cycles of typical loading. It was also field tested on adult orthoses over a minimum of a 12-month period with the most vigorous of walkers. In neither test did excessive play develop. The mechanical properties of the joint were established using tests advocated in the British Standard on testing lower limb orthosis knee joints. These showed the joint had structurally equivalent performance to a successful reciprocal walking orthosis hip joint, and that the mode of failure was essentially ductile in nature. Production development of the joint is now being undertaken. PMID- 11411006 TI - Shoes in the cupboard: the fate of prescribed footwear? AB - Studies show that patient dissatisfaction with prescribed footwear results in low usage and that the cause is the system that provides it. The aim of this study was to compare referrer and patient perceptions of a multidisciplinary footwear clinic (MDFC) and a traditional surgical appliance clinic (SAC). Referring consultants completed a questionnaire for every referral (28 referred to the MDFC and 27 to the SAC) during a six-month period. Structured interviews were designed to gather the data. The results showed that referrers to both groups indicated that they lacked the time and knowledge to give advice about retail footwear. The entire MDFC group but only 18 (66.6%) of patients in the SAC group had the opportunity for discussion about footwear before the prescription took place. Some 17 (62.9%) in the SAC group reported problems with the footwear but only 11 (40.7%) of this group were informed what to do if problems occurred. Both groups demonstrated an association between the patient's attitude, their perception of improvement in their feet and overall satisfaction. These factors seemed to have an impact on the usage of the prescription footwear with 13 (48.1%) of the SAC group using other footwear compared with 2 (7.2%) in the MDFC group. This study adds to the suggestions made in other studies that there is a need to review the SAC system and identifies key areas that influence patient satisfaction. PMID- 11411007 TI - Effectiveness of audio-biofeedback in postural training for adolescent idiopathic scoliosis patients. AB - The possibility of using learned physiological responses in control of progressive adolescent idiopathic scoliosis (AIS) was investigated. Sixteen (16) AIS patients with progressing or high-risk curves (Cobb's angle between 25 degrees and 35 degrees at start and reducible by lateral bending) were fitted with a device with tone alarm for poor posture. In the first 18 months of application, 3 patients defaulted and 4 showed curve progression > 10 degrees (2 changed to rigid spinal orthoses and 2 underwent surgery). The curves for the other 9 patients were kept under control (within +/- 5 degrees of Cobb's angle) and 5 of them have reached skeletal maturity and terminated the application. The remaining 4 patients were still using the devices until skeletal maturity or curve progression. The curve control rate was 69%. A long-lasting active spinal control could be achieved through the patient's own spinal muscles. Nevertheless, before the postural training device could become a treatment modality, a long term study for more AIS patients was necessary. This project is ongoing in the Duchess of Kent Children's Hospital, Sandy Bay, Hong Kong. PMID- 11411008 TI - Lower limb amputation. Part 1: Amputation methods--a 10 year literature review. PMID- 11411009 TI - ParaWalker ambulation for adult tetraplegic patients: two case reports. PMID- 11411010 TI - Congenital nephrotic syndrome: a clinico-pathologic study of thirty children. AB - BACKGROUND: Congenital nephrotic syndrome is an uncommon disorder that may be caused by several diseases. These may be inherited, sporadic, acquired or part of a general malformation syndrome. METHODS: We reviewed the clinical characteristics, pathologic findings, and results of medical management in 30 infants who presented to Jordan University Hospital with congenital nephrotic syndrome in the years 1989 to 1999. RESULTS: Most patients (80%) had parents who were consanguineous. Most patients (80%) were born premature, with an average gestational age of 36 weeks. Most infants (77%) presented the nephrotic syndrome in the first three months of life and 26 (87%) had significant growth retardation. Twenty-five verified episodes of serious bacterial infections occurred in 18 patients. Antibiotic therapy however was successful in all these episodes. Light microscopy of the renal biopsies was consistent with the Finnish type of congenital nephrosis in most patients (83%). Chronic renal insufficiency developed in 17, and five of them needed chronic peritoneal dialysis. Most patients were given albumin transfusion and diuretic therapy especially during episodes of severe edema. Captopril alone or in combination with ibuprofen was given to eight patients, but without a response in any of them. All patients died before the age of 5 years. Most deaths occurred at an average age of 15 months (range 1-60). CONCLUSION: The Finnish type of congenital nephrosis was the most common type in our patients, most of whom died within a few months of the onset of disease. In the developing countries, the management of patients with congenital nephrosis may have to be different from that in the developed countries in view of the high cost of medical management, poor outcome, high risk of serious complications, and high mortality rate. PMID- 11411011 TI - The outcome of non-selective vs selective nitric oxide synthase inhibition in lipopolysaccharide treated rats. AB - Nitric oxide (NO), generated by inducible nitric oxide synthase (NOS) following lipopolysaccharide (LPS) administration, produces renal failure through autoinhibition of glomerular endothelial NOS activity. Preadministration of selective iNOS inhibitors abolishes this effect. Although nonselective NOS inhibitors further decrease GFR, current clinical trials investigate the effect of nonselective NOS inhibition in septic patients. The goals of our study were to determine whether treatment with selective NOS inhibitors can reverse the decrease in GFR in LPS treated rats with already established renal failure and to define the outcome of LPS treated rats following nonselective NOS inhibition. Four hours following the administration of LPS (4 mg/kg), we measured creatinine clearance (CrCl) before and after the administration of either L-NIL (selective iNOS inhibitor, 3 mg every 20 minutes) or saline. Selective iNOS inhibition attenuated the decrease in blood pressure [ CONTROLS: 105 +/- 6 to 98 +/- 5, LPS: 92 +/- 5* to 83 +/- 4*, LPS + L-NIL: 88 +/- 6* to 94 +/- 6 mm Hg; *p < 0.05, vs controls (n = 6)], and reversed the decrease in GFR after LPS [ CONTROLS: 2.21 +/ 0.13 to 2.07 +/- 0.11, LPS: 0.82 +/- 0.18* to 0.66 +/- 0.22*, LPS + L-NIL: 0.76 +/- 0.15* to 1.86 +/- 0.15 ml/min; *p < 0.05 vs controls (n = 6)]. We next studied the effect of complete non-selective NOS inhibition (L-NAME 200 mg, 2 hours after LPS) on LPS treated rats. All (6/6) animals treated with both LPS and L-NAME died within 2 hours following LPS, while rats treated with either LPS, L NAME, or LPS + L-NIL survived. Histologic studies performed in all experimental groups were unremarkable. Overnight mortality was studied using smaller doses of L-NAME. All LPS + L-NAME (10/10) and 1/10 LPS treated rats died. L-NAME, control, and LPS + L-NIL animals survived. The characteristic histologic findings in LPS + L-NAME rats were diffuse ischemic changes, most importantly acute myocardial infarction. IN CONCLUSION: Selective iN-OS inhibition might prove to have clinical application as it prevents the decrease in GFR following LPS, even after renal failure is established. Treatment with a non selective NOS inhibitor in septic patients should be reconsidered. PMID- 11411012 TI - Immunoglobulin A nephropathy associated with hepatitis A virus infection. AB - Renal involvement in association with nonfulminant hepatitis A virus (HAV) infections have been reported infrequently and when present have been characterized by mild proteinuria, microscopic hematuria and slight urinary sediment abnormalities. Acute renal failure and nephrotic syndrome are extremely rare complications. This report describes a case of acute renal failure and nephrotic syndrome following a nonfulminant course of HAV infection with biopsy proven immunoglobulin A nephropathy. The cause of acute renal failure in this case is primarily due to tubulointerstitial nephritis secondary to pigment toxicity. PMID- 11411013 TI - Idiopathic polymyositis and glomerulonephritis. AB - We report a 28-year-old male who presented with a clinical picture compatible with idiopathic polymyositis and nephrotic-range proteinuria. Muscle biopsy confirmed the diagnosis of polymyositis and a diagnostic renal biopsy demonstrated IgM mesangial glomerulonephritis. Following a short-course of prednisone, both the myositis and proteinuria resolved. Glomerulonephritis associated with idiopathic polymyositis is rare; however, since it appeared to respond to corticosteroid therapy concomitant with the improvement in the myositis, it was likely an associated immunological complication. PMID- 11411014 TI - Cardiac and intestinal amyloidosis in a renal transplant recipient with familial Mediterranean fever. AB - In Turkey, familial Mediterranean fever (FMF) is an important cause of nephrotic syndrome and endstage renal disease due to renal deposition of AA type amyloid. We report a case of living-related donor renal transplant recipient with FMF and renal AA type amyloidosis, who died of progressive heart failure due to cardiac involvement. The patient also had intractable diarrhea caused by biopsy-proven intestinal amyloidosis. The patient was on 1 mg/day colchicine. Although he was attack-free throughout the post-transplant period, intestinal and clinically significant cardiac amyloidosis, which implied the presence of sustained inflammation and continuing amyloid deposition, appeared three years after renal transplantation. Cardiac deposition of AA amyloid may cause clinically significant heart disease, leading to cardiovascular mortality after renal transplantation for end-stage renal disease in FMF patients. PMID- 11411015 TI - Nodular macroglossia with combined light chain and beta-2 microglobulin deposition in a long-term dialysis patient. AB - We describe a case in which nodular macroglossia, a very rare type of tongue involvement, was associated with the co-deposition of lambda light chain and beta 2 microglobulin fibrils in the tongue. The combined presence of two different amyloid fibrils did not lead to a more unfavourable clinical outcome. We believe that both these features often remain underdiagnosed and are in fact more frequent than reported. A careful clinical examination of the tongue together with serum immunofixation should be routine in all patients with dialysis-related amyloidosis in order to investigate the prevalence and type of tongue involvement and to rule out other types of amyloidosis. In all cases of suspected mixed amyloidosis, immunohistochemical characterization of fibrils should be carried out by electron microscopy. PMID- 11411016 TI - Non tuberculous mycobacterium peritonitis in continuous ambulatory peritoneal dialysis. AB - A forty-five-year old Saudi lady who had been on Continuous Ambulatory Peritoneal Dialysis (CAPD) for three years, was admitted with a clinical picture of night fever, sweating, weight loss and turbid peritoneal fluid (PF). The PF had a high cell count, predominantly neutrophils. This condition failed to respond to a standard vancomycin-gentamycin treatment, and acid fast bacilli (AFBs) were stained from the PF. The patient was commenced on antituberculous treatment and the Tenckhoff catheter was removed. She was shifted to haemodialysis and recovered fully. Later, the AFBs were identified as a strain of mycobacterium fortuitum. PMID- 11411017 TI - Prevention and management of late renal allograft dysfunction. AB - Chronic allograft nephropathy is the leading cause of return to dialysis after transplantation. At present, no efficient therapies are available to modify the natural history of chronic allograft nephropathy, mainly because of difficulties in designing prospective clinical trials to prevent or treat this entity. The main risk factors for chronic allograft nephropathy, the utility of protocol biopsies in the design of trials, and different strategies to prevent or treat this problem are reviewed. PMID- 11411018 TI - TT virus infection in end-stage renal disease (ESRD). AB - TT virus is a novel hepatitis-associated DNA virus that has been provisionally designated as transfusion-transmitted virus (TTV). Infection by TTV has been frequently demonstrated in humans. Few reports, however, have been published on the epidemiology of TTV infection in patients with ESRD. The prevalence of TTV in patients undergoing maintenance dialysis varies widely and various demographic, virologic or clinical features can explain these differences; in some series, the prevalence of TTV in HD was significantly higher than in control groups. Blood transfusion requirement and nosocomial transmission of TTV within dialysis units seem to be important in the diffusion of TTV in the HD setting; however, other routes of TTV acquisition may play a role. The clinical significance related to the presence of TTV in HD population remains unclear; it is possible that TTV may aggravate liver disease caused by hepatitis C virus (HCV) infection. The natural history of TTV in dialysis is an area of avide research, since TTV has a propensity for establishing long-term infection in HD patients. The possibility that TTV causes pathological changes outside the liver cannot be ruled out. Further studies are in progress to understand the natural history and the epidemiology of TTV infection in patients with ESRD. PMID- 11411019 TI - What can nephrologists learn from epidemiology? AB - During the last decade, epidemiology has played an increasing role in the study of renal diseases. In the fields of drug-nephrotoxicity and occupational renal failure, epidemiological research has made an important contribution. In recent years, the specific role of epidemiology in outcome research has gained recognition. Epidemiology is based on observations under real-life conditions using a representative sample of renal patients. Methods range from simple clinical observations and cross-sectional studies to case-control and prospective cohort studies. Contributions and limitations of the different epidemiological approaches are illustrated using the example of the nephrotoxicity of 5 aminosalicylic acid in patients with inflammatory bowel disease. PMID- 11411020 TI - Bioethics in nephrology: guidelines for decision-making in Italy. AB - In 1998, the Italian Ministry of Health urged the Italian Society of Nephrology (SIN) to draft recommendations for uniform decision-making procedures in the field of dialysis. In accordance with the Ministry's specifications, at the time the Society approved diagnostic protocols regarding clinical questions, but without fully recognizing the multiple issues arising in complex medical situations where dialysis is likely to go on for a long time. The authors propose some guidelines for ethical conduct and their application, with special regard to medico-legal dilemmas, together with the principles of clinical guidelines that would further improve the "perceived quality" of chronic dialytic treatment. PMID- 11411021 TI - Nephrin mRNA regulation by protein kinase C. AB - BACKGROUND: Mutations in the recently cloned NPHS1 gene result in congenital nephrotic syndrome of the Finnish type (CNF). The protein product of NPHS1, nephrin, is expressed uniquely in kidney glomerular podocytes, and is the first true component of the interpodocyte slit membrane. The precise functions of nephrin remain unknown, but the presence of several tyrosine residues in the intracellular domain suggest a role in signalling. We searched for nephrin expressing cell line for use in signal transduction studies and also characterized the main features of calcium signalling in nephrin-deficient cultural glomerular epithelial cells. METHODS: We used A293 cell line, found to naturally express nephrin, as well as cultured CNF glomerular cells using reverse transcription PCR, immunocytochemistry and intracellular Ca2+ measurements. RESULTS: Phorbol-12-myristate-13-acetate significantly upregulated the nephrin expression in A293 cells, while no change was found after treatment with additional stimulants for other main signalling pathways, e.g. okadaic acid, lysophosphatidic acid, bradykinin, angiotensin II (ANG II) and arginine vasopressin (AVP). No changes in basal or ANG II- or AVP-stimulated intracellular Ca2+ fluxes in CNF glomerular cells were observed. CONCLUSIONS: Protein kinase C may be the key intracellular signalling system in the regulation of nephrin. PMID- 11411022 TI - Effect of organizational change on the individual employee. AB - Increasing market pressures force companies to implement drastic organizational changes in order to remain competitive. Budget decreases, reduction efforts, and similar changes create significant morale and job-satisfaction concerns. This study assesses the effects of budget reductions and other organizational changes on the morale of hospital employees. A survey dealing with employee perceptions of stress, workload, and performance was given to hospital employees. Not surprisingly, the survey found that morale problems resulted from the organizational changes. Employees' gender and job classification showed little significant effect on the survey results, while respondents' length of employment with the organization influenced the results slightly. The findings provide information useful for dealing with challenges of employee satisfaction, morale, and trust during times of budget limitations. PMID- 11411023 TI - Marketing in today's health care environment. AB - The application of health care marketing is seen as a relatively recent phenomenon. Marketing is discussed as a sophisticated managerial tool that includes five critical components: product, price, place, promotion, and partners. The triumvirate of health care decision makers (i.e., patients, insurance organizations, and employers) are examined vis-a-vis these five components. PMID- 11411024 TI - Managing clinical data in an electronic environment. AB - Clinical data has moved to the forefront as a valuable resource to enhance patient care and measure organizational performance. These data, most of which is now captured electronically, must be assessed for value and appropriateness. This article examines a data quality management model and its use in the initiation of computerized patient record (CPR) projects. PMID- 11411025 TI - Teamwork or interdepartmental cooperation: which is more important in the health care setting? AB - A survey of 75 nursing department employees was conducted to assess the relative importance of across-department and within-team cooperation on workplace outcomes. As compared with within-team cooperation, across-department cooperation is more positively associated with procedural justice, interpersonal justice, satisfaction with supervisor feedback, supervisory rating, job satisfaction, and organizational commitment. Across-department cooperation is more negatively associated with role ambiguity, role conflict, role overload, job tension, and job withdrawal intentions. No significant correlational differences are noted for either distributive justice or politics. Type of cooperation also was analyzed using hierarchical regression, and more variance was explained in across department cooperation than within-team cooperation by organizational variables. Based on these results, it may be more important for the health care manager to attend to issues of interdepartmental cooperation rather than to internal team dynamics. PMID- 11411026 TI - Being good and doing good: the culture of professionalism in the health professions. AB - The characteristics of being good and doing good are discussed in the current societal context of distrust, cynicism, and decreasing integrity and commitment. Health professionals choose being good and doing good as a career. Many reasons are given as to why professionalism has declined in the health professions: "the system," decreased individual rewards and personal satisfaction, and the loss of focus on the patient. The author suggests three levels of aggressive intervention -the professional organization, the organization one works for, and the individual--to revitalize good citizenship and professionalism. PMID- 11411027 TI - The manager and equipment decisions: is that in the capital budget? AB - Although any decision to purchase a piece of capital equipment involves a number of the organization's functions, the department manager has some significant responsibilities in the selection and acquisition of capital equipment. Except for unavoidable replacement of essential equipment that fails unexpectedly, capital purchases must ordinarily be planned in advance through the annual budgeting process. The department manager is ordinarily the organization's primary source of information in major capital expenditure projects; therefore, it is essential that the manager follow a logical process for identifying and evaluating alternative equipment choices and perform a consistent economic analysis of the alternatives. PMID- 11411028 TI - A manager asks: tardiness and flexing--it's about time. PMID- 11411030 TI - The medical lessons of science fiction. PMID- 11411031 TI - Medical ethics through the Star Trek lens. PMID- 11411032 TI - In memoriam: Elizabeth Sewell. PMID- 11411033 TI - From cyborg fiction to medical reality. PMID- 11411034 TI - Microscopic doctors and molecular black bags: science fiction's prescription for nanotechnology and medicine. PMID- 11411035 TI - A brief history of stories. PMID- 11411036 TI - "Not a virus, but an upgrade": the ethics of epidemic evolution in Greg Bear's Darwin's Radio. PMID- 11411037 TI - Semecarpus anacardium Linn. nuts--a boon in alternative medicine. AB - In alternative medicine, medicinal plant preparations have found widespread use particularly in the case of diseases not amenable to treatment by modern methods. Chemical and phytochemical analyses of Semecarpus anacardium nut reveal the presence of biflavonoids, phenolic compounds, bhilawanols, minerals, vitamins and amino acids. A variety of nut extract preparations from this source are effective against many diseases, viz. arthritis, tumours, infections etc. and non-toxic even at high dose of 2000 mg/kg. However understanding of the mechanism of the pharmacological action of S. anacardium nut can be greatly aided by the isolation of its active principle from the nut and determination of the structure-function relationship. Also, the potent curative effect of S. anacardium nut extract against human ailments need to be verified by controlled clinical studies. PMID- 11411038 TI - Human granulosa cells in vitro: characteristics of growth, morphology and influence of some cytokines on steroidogenesis. AB - Granulosa cells (GCs) were characterised morphologically by light and electron microscopy. The steroidogenic capability of GCs in vitro was estimated by radioimmunoassay (RIA): oestradiol (E2), progesterone (P) and androstenedione (A) secreted into the culture medium were measured. The influence of several culture media and anchorage of the cell either to plastic vessels (monolayer) or to collagen fibrils (in gel) were studied. As the various culture media were assayed with regard to their suitability for IVF, it was found that Ham's F10 is quite satisfactory (in agreement with other observations on embryo cultures). A chemically defined medium BM 86 was found to be inadequate. In addition to the two cell types which are known, a third cell type which can perform efficient aromatisation (E2 production) in vitro is characterised here. The influence of cytokines/growth factors (GF) like insulin-like GF (IGF-1), epidermal GF (EGF), platelet-derived GF (PDGF) and fibroblast GF (FGF) on steroidogenesis was tested either alone or with human chorionic gonadotrophin (hCG). Except for oestradiol (E2) from early GCs, hCG generally stimulated progesterone (P) and E2 secretion. EGF by itself enhanced the secretion of P but not of E2. EGF did not affect hCG stimulation of P, but reduced that of E2. In contrast, in pre-ovulatory GCs IGF-I reduced the stimulatory effect of hCG on both E2 and P. In early GCs IGF-I potentiated hCG stimulation of P. In early GCs, neither hCG nor IGF-I nor a combination of IGF-I with hCG had any effect on E2 production. PMID- 11411039 TI - Cell culture of biopsied endometriomas after danazol/hormonal therapy: a study of growth features and fertility effects. AB - The growth features of cells from endometriomas biopsied from patients who had been treated with Danazol or with hormones have not been studied in vitro. Danazol is a versatile drug and despite its recognised efficacy in controlling endometriosis, little is known about is cytotoxicity and mechanism of action. Culture of the biopsied endometriomas permitted qualitative cytotoxic assessments by way of comparison with cultured normal uterine endometrial cells treated in vitro with Danazol. The growth characteristics were studied in monolayer and collagen gel cultures. Cytopathology was characterised by light and electron microscopy. Since endometriosis is associated with infertility in women, data from in vitro fertilisation (IVF) were analysed with respect to treatment modalities as compared with cases suffering fallopian occlusion. Danazol reduced pregnancy chances. Two factors may be responsible: (a) altered follicle development resulting in poor oocyte quality (b) reduced nidation because of long lasting endometrial cytotoxicity. The experimental findings reported here support the latter explanation. Consequently, Danazol therapy should be conditional; patients wishing to achieve pregnancy should preferably receive hormonal therapy. PMID- 11411040 TI - A new continuous cell line from larval hemocytes of Spodoptera litura (F.). AB - A new cell line has been established from larval hemocytes of the moth, S. litura (tobacco cut worm). It took 147 days to form a monolayer and one year for the first 17 passages. At present, the culture is at 86th passage level and is designated NIV-SU-1095. Three cell types could be distinguished, viz. plasmatocytes (53%), prohemocytes (36%) and granular hemocytes (11%). The chromosome number was very high, 74% metaphase cells showed more than 100 chromosomes. The cells could be cryopreserved. The cells were susceptible to the baculoviruses, Autographa californica nuclear polyhedrosis virus and S. litura nuclear polyhedrosis virus (SLNPV). Plaques could be observed on 7th post infection day with SLNPV. Six cloned cell lines have been developed of which clone II-1F was more sensitive to both the baculoviruses compared to the original cell line. PMID- 11411041 TI - Suppression of Ehrlich ascites tumor growth by immunization with ganglioside GT1b of its origin, its IgM antibody or anti-idiotype of the anti-GT1b IgM. AB - Ehrlich tumor expresses the ganglioside GT1b. The plasma of mice with Ehrlich ascites tumor burden also contains GT1b. The structural identity of plasma GT1b was ascertained by a series of enzymatic degradation and mass spectral analysis. Mice were vaccinated with purified plasma GT1b admixed with Freund's adjuvant (FA). Sixty nine percent suppression of Ehrlich ascites tumor growth was observed in vaccinated mice. The suppression was dose-dependent. It is hypothesized that the tumor growth-suppression is a result of immune response to GT1b Humoral immune response to GT1b was demonstrated by passive hemagglutination assay of the sera of vaccinated mice. To test the hypothesis, the mice were administered with rabbit polyclonal anti-GT1b IgM antibody in varying doses and challenged with Ehrlich tumor. A significant reduction in tumor growth (65%) was observed in mice administered with anti-GT1b IgM antibody. Again, the suppression was dose dependent. To verify further, another batch of mice was immunized with anti idiotypic antibodies to rabbit anti-GT1b IgM raised in rat. The polyclonal anti idiotype antibody is expected to carry the structural image of GT1b. In a dose dependent manner, a maximum of 82% suppression of tumor growth was observed in mice immunized with the anti-idiotype antibody. This observation further strengthened the hypothesis that ganglioside mediated suppression of tumor growth may be a result of immunogenicity of the target ganglioside. This was also supported by positive reaction of the sera of anti-idiotype vaccinated mice with both anti-idiotype antibody and ganglioside GT1b in passive hemagglutination assay. The results favour the therapeutic potential of immunogenic tumor associated gangliosides. PMID- 11411042 TI - Studies on safety aspects of contraceptive Magainin-A in rabbits. AB - We have previously reported the contraceptive potential of Magainin-A in rats and rabbits under in vitro and in vivo condition. In this report we evaluated the effect of Magainin-A on the structural organisation of vaginal epithelium in rabbits. The effect of this compound on the erythrocytes and its rate of absorption and clearance from systemic circulation was also studied. The effective contraceptive dose of Magainin-A (1 mg) when administered intra vaginally for five consecutive days did not induce any structural or morphological abnormalities in vaginal epithelial cells. No adverse effect was observed on the erythrocytes. The rate of Magainin-A absorption and clearance from the circulation was found to be rapid. These results suggest that Magainin-A may be used as a safe intra-vaginal contraceptive compound in future. PMID- 11411043 TI - Protein tyrosine kinase activity in human malaria parasite Plasmodium falciparum. AB - Protein tyrosine kinases (PTKs) are believed to be implicated in the parasite growth, maturation and differentiation functions. Protein tyrosine kinase activity was found to be distributed in all the stages of P. falciparum parasite maturation. Membrane bound PTK activity was found to be increased during maturation process (ring stage to trophozoite stage) in chloroquine sensitive strains. In vivo conversion of the schizont stage to ring stage via release of merozoites was associated with a decrease in PTK activity. Chloroquine inhibited the membrane bound PTK activity in a dose dependent manner (IC50 = 45 microM). Kinetic studies show that chloroquine is a competitive inhibitor of PTK with respect to peptide substrate and noncompetitive with respect to ATP indicating that chloroquine inhibits PTK activity by binding with protein substrate binding site. The results suggest that maturation of malaria parasite is related to PTK and inhibition of this activity by chloroquine could provide a hypothesis to explain the mechanism of action of chloroquine. PMID- 11411044 TI - Effect of cortisol on testis of freshwater fish Notopterus notopterus (Pallas). AB - Cortisol (20, 40 and 60 micrograms/fish for 10 days) treatment caused an increase in testicular-somatic index (TSI) in immature N. notopterus whereas in mature fish no change from that of controls was observed. Histology of testis indicated that spermatogenetic activity was activated in immature fish while it was inhibited in mature fish. Testicular cholesterol exhibited a similar response. The results indicate that cortisol inhibits spermatogenesis during mature phase while it promotes spermatogenesis during immature phase of the reproductive cycle in N. notopterus. PMID- 11411045 TI - In vitro stimulation of chick brain lipid peroxidation by aluminium, and effects of tiron, EDTA and some antioxidants. AB - In vitro effect of aluminium (Al) on Fe(2+)-induced lipid peroxidation (LPX) in various subcellular fractions from the cerebral hemispheres (CH) of 7- and 30-day old chicks was studied. Stimulation of Fe(2+)-induced LPX by Al was observed to be the highest in microsomal fraction. The magnitude of elevation of Fe(2+) induced LPX in various subcellular fractions of brain showed age related variation. Of the six chemicals tested for their influence on Al-induced lipid peroxidation, both doses of 1,2-dihydroxybenzene-3,5-disulphonic acid disodium salt (Tiron), ethylene diamine tetra acetic acid disodium salt (EDTA), and ascorbic acid prevented the Al-induced LPX in crude homogenates of the CH, whereas only at a higher dose inhibition by 1,4-diazabicyclo (2.2.2.) octan (DABCO) was observed. On the contrary, mannitol and dimethyl sulfoxide did not inhibit the induction of LPX by Al in crude homogenate. The effect of test chemicals on Al-induced LPX in both the ages of chick tissue was almost similar. The results suggest that Al further augments Fe(2+)-induced LPX in various compartments of the cell due to generation of free radicals. The results also showed that Tiron, EDTA and antioxidants, such as ascorbic acid and DABCO can prevent LPX induced by Al. PMID- 11411046 TI - Successful propagation in vitro of apple rootstock MM106 and influence of phloroglucinol. AB - Successful in vitro propagation of clonal apple rootstock MM106 was achieved by culturing axillary buds on MS basal medium with BAP (1 mg/L), GA3 (0.5 mg/L) and IBA (0.1 mg/L). Use of liquid medium (LM) in initial cultures reduced phenol exudation to a greater extent and gave maximum sprouting percentage when transferred to solid MS medium. Phloroglucinol (PG) did not enhance sprouting of buds but increased the rate of multiplication when added in the medium. Maximum number of shoots were obtained when MS medium was supplemented with BAP (0.5 mg/L), GA3 (1 mg/L), IBA (0.1 mg/L) and PG (100 mg/L). For rooting, in vitro regenerated shoots were placed in IBA (30 mg/L) for 3 hr and transferred to solidified auxin free medium. Rooting was recorded in about 80% of shoots. Inclusion of PG in rooting medium was not beneficial but shoot cultures grown in its presence gave higher rooting percentage. Rooted plantlets showed about 70% survival rate in potting mixture of sand:soil:perlite (1:1:1). PMID- 11411047 TI - Somatic embryogenesis in Vigna radiata (L.) Wilczek. AB - Somatic embryogenesis was achieved from immature cotyledon derived callus of mungbean, V.radiata (L.) Wilczek in MS liquid medium. Embryogenic callus was induced on MS medium with NAA (5 mg/L). Differentiation of somatic embryos was observed when embryogenic callus was transferred to MS liquid medium containing 2,4-D (1.5 mg/L) and L-proline (50 mg/L). The torpedo shaped embryos were transferred to MS liquid medium with BAP and ABA (1 mg/L each) for maturation and germination. Fifty per cent of torpedo shaped embryos were converted into tiny plants (8-9 plants out of 17) after one week of culture. The germinated embryos were isolated and transferred to MS half strength basal (solid) medium for further development. PMID- 11411048 TI - Influence of metham sodium on suppression of collar rot disease of peanut, in vitro antibiosis, siderophore production and root colonization by a fluorescent pseudomonad strain FPO4. AB - A hydroxamate type siderophore producing fluorescent Pseudomonas strain, isolated from the rhizoplane of paddy root showing plant growth promoting activity, exhibited a decreased in vitro antibiosis, production of siderophore and suppression of collar rot in presence of metham sodium. Use of herbicide had a detrimental effect on the plant growth promoting activity of this organism. The multiple drug resistant mutant strain derived from this rhizobacteria colonized the roots, but the herbicide application had a negative effect on their population. HPLC analysis of the siderophore showed five peaks of which the peak number three confirmed the antifungal activity. PMID- 11411049 TI - Carbendazim generates symplasts in rat spermatogenic clones. AB - In order to find non-microtubular targets in the seminiferous epithelium for the fungicide and reproductive toxicant carbendazim, it was administered to 90 days old male Wistar rat in a single bolus dose of 400 mg/kg body weight through an oral intubation. A parallel control group was maintained. Rats were sacrificed 48 days after the treatment and the testes were analysed for histopathological changes adopting routine histological methods, when symplasts were localised. The maximum diameter of five largest symplasts was measured, and the number of nuclei in these symplasts was also determined. As it is known that symplasts of spermatogenic cells are produced due to opening up of the intercellular bridges between cells in a clone consequent upon disruption of actin microfilaments, the present study shows that actin microfilaments would also be targets in the seminiferous epithelium for carbendazim toxicity. PMID- 11411050 TI - Effect of titanium exposure on embryonic development during pre-implantation period in rats. AB - Doses of titanium trichloride (1/10th and 1/5th of LD50) were administered once and daily to pregnant rats to assess their effect on embryonic development. 1/5th dose of TiCl3 administered once orally on 1st, 2nd and 3rd day post-coitum. Similarly 1/10th of LD50 was administered daily. Results revealed that 1/10th LD50 dose of TiCl3 was more effective during pre-implantation period as number of 4 and 8-celled embryos decreased as compared to 1/5th. Delayed hatching of the blastocysts on day 5 was registered in TiCl3 treated dam. PMID- 11411051 TI - Generation and characterization of monoclonal antibodies to adenovirus. AB - Monoclonal antibodies (MoAbs) were generated following immunization of Balb/C mice with adenovirus type 5 grown in Hep2 cell line. Six clones reactive to hexon antigen of the virus were stabilized, of which 4 had mu-heavy chain specificity and 2 were of gamma-heavy chain type. Three of the clones (ADV-1, ADV-3 and ADV 5) had a high ELISA reactivity to the hexon antigen but exhibited differential specificity to the adenovirus types tested. In Western blotting, ADV-1 and ADV-3 reacted with all the adenovirus types tested (types 3,4,5,7 and 8) with reactions at 116 kDa region (hexon antigen), in addition, ADV-3 also had reactivity at 80 kDa region, the penton antigen. Reactivity to these adenoviral types by the 2 MoAbs was demonstrable by dot ELISA. ADV-5 had a type specific reaction only to adenovirus type 5 in dot ELISA with specificity in the hexon region in Western blotting. The reactivity of these 3 clones was not observed to the normal Hep2 tissue culture antigens and to the 3 enteroviruses tested (polio, coxsackie A9 and echo 4). PMID- 11411052 TI - Racial divergence in abdominal bristles among parental races and newly evolved cytoraces of nasuta-albomicans complex of Drosophila. AB - Cytoraces are the products of interracial hybridization between Drosophila nasuta nasuta and D. nasuta albomicans. These races differ from their parents in the chromosome composition, mating preference, certain fitness phenotypes and also a few morphophenotypic traits. Now, these cytoraces are passing through 330 generations. Racial divergence in the 4th and 5th abdominal bristles among the parental races and the newly evolved cytorace 1 and 2 is reported. The results revealed that the parental races have more number of bristles than newly evolved cytoraces. Thus, these cytoraces are evolved/evolving with reduced abdominal bristle number and better fitness. PMID- 11411053 TI - Response of Java citronella (Cymbopogon winterianus Jowitt) to toxic heavy metal cadmium. AB - Cadmium at 200 mg kg-1 soil and above concentrations was fatal as growth was inhibited ultimately leading to death of Java citronella (Cymbopogon winterianus Jowitt.). The surviving plants at 50 and 100 mg kg-1 treatments also exhibited pronounced retardation of growth and biomass yield. There was considerable reduction in the level of essential oil in herbage and oil quality deteriorated. Cadmium accumulation profile showed that highest accumulation was in root, followed by stem, leaf sheath and leaf. Very high accumulation in root for higher doses appeared to be the reason for fatality. PMID- 11411054 TI - Influence of growth regulators and explant type on in vitro shoot propagation and rooting of red sandal wood (Pterocarpus santalinus L.). AB - In vitro shoot regeneration in Pterocarpus santalinus L. was achieved when detached cotyledons from in vitro germinated seedlings were cultured on MS medium containing NAA (0.1 mg/L), BA (1 mg/L) and kinetin (1 mg/L). The regenerated shoots rooted on 1/4 strength MS medium with IAA (1 mg/L) and the fully developed plantlets were successfully established in the soil. PMID- 11411055 TI - In vitro adventitious shoot formation from leaf cultures of Clerodendrum inerme (L) Gaertn. AB - In vitro adventitious shoots (about 28) of Clerodendrum inerme were regenerated from leaf segments on MS medium containing BA (4 mg/L). These shoots developed directly from the leaf explants without callusing after 5 weeks. Leaf explant when cultured in MS medium containing BA (2 mg/L) and NAA (0.5 mg/L) developed compact callus that became nodular and regenerated shoots (about 50) after 5 weeks. The in vitro developed shoots were rooted in MS medium supplemented with IAA (2 mg/L). The hardened plantlets were successfully established in the field with 90% survival. PMID- 11411056 TI - [Pain in elderly patients and aspects of its therapy]. AB - Treatment of pain in elderly patients can meet with diagnostic problems, namely in those with cognitive disorders, as well as with problems concerning pharmacokinetic and pharmacodynamics brought about by the advanced age. Our article presents an overview on the basic differences in the diagnostics and treatment of pain, on the therapeutic risks, and on other interrelations, which have to be considered in the therapy of elderly patients. The review is supplemented with results of our examination on the rate of analgesic treatment, spectrum of analgesics in use in the population of patients older than 75 years. About one quarter of the population use analgesics regularly, the order of most frequently prescribed analgesics is: ibuprofen 38%, diclophenac 24%, tiaprophenic acid 14%, tramadol 8%, indometacine 4%. Included were also data concerning the quality of life of patients with pain and information about the relations among patients and the curing personnel where obtained by our own inquiry. Patients concluded that pain was better tolerated at home, administration of drugs in tablets revealed to be most satisfactory, all information, namely from medial doctors, were welcome. Treatment of pain improved the quality of life in 1/3 of patients; one half of them considered it as successful. Personnel also asked for better professional information, though they had good of knowledge on the evaluation and documentation of pain and the principles of pharmacological treatment. Failure of treatment evokes in 69% of the personnel the feeling of impotency, in 41% affection of unsatisfactoriness, in 20% a tension, in 13% feeling of failing, in 18% depression and frustration. Reconciled with failure is 6% of the curing personnel. PMID- 11411057 TI - [Nonvariceal acute upper digestive tract hemorrhage--therapeutic approach]. AB - Nonvariceal acute upper gastrointestinal hemorrhage continues to be a frequent cause for hospital admission, consumes considerable financial resources and belongs to diagnoses associated with significant morbidity and mortality. Despite the progress in endoscopic and intensive care therapies the mortality remained unchanged. It results from the increasing number of high risk patients, namely the older ones with significant comorbidity. The cornerstone of therapeutical success is considered to be the adequate hemodynamic and ventilatory stability, initiation of pharmacotherapy with proton pump inhibitors and eventually somatostatin, and in particular, the early endoscopy using modern methods of hemostasis. In case of two unsuccessful endoscopic sessions the surgical approach is justified. PMID- 11411058 TI - [Osteopenia and osteoporosis in diabetic women over 40 years of age]. AB - BACKGROUND: The aim of this study was to find the prevalence of osteopenia and osteoporosis in postmenopausal diabetic females and its association with the diabetic compensation and angiopathic complications. METHODS AND RESULTS: Bone mineral density was measured in 156 diabetic females, 44 of them had type 1 diabetes mellitus (mean age 53 +/- 8.9 years) and 112 type 2 (mean age 60 +/- 9.2 years). In the lumbar spine and proximal femur the prevalence of osteoporosis varied between 9-11% and 18-20% respectively. The respective prevalence of osteopenia amounted 30-36% and 34-48%. No statistically significant differences were found between type 1 and type 2 diabetes. Any association with diabetic compensation and the diabetic microangiopathic complications was not found. Only in females with osteoporosis the prevalence of cardiovascular heart disease was higher than in those with normal bone mineral density. CONCLUSION: The prevalence of osteoporosis and osteopenia in postmenopausal diabetic women is similar to those in the common population of the respective age. PMID- 11411059 TI - [Metformin in the treatment of type 1 diabetics--a placebo controlled study]. AB - BACKGROUND: Metformin is popular for it's complex mechanism of action in treatment of the type 2. diabetes. The effect in type 1. diabetes is studied less frequently. The aim of our open, prospective, placebo controlled study was to assess the effect of metformin in poorly controlled diabetic patients type 1 with high insulin requirements. METHODS AND RESULTS: In the group comprised of 19 type 1 diabetic patients the insulin resistance was assessed by hyperinsulinemic euglycemic clamp and indirect calorimetry at the beginning of the study (B), 3 months later when metformin in the dose of 2 x 850 mg was added to existing insulin therapy (M) and after 3 months of placebo therapy (P). In the same time intervals the other parameters were measured. Wilcoxon test was used for statistic analysis. All results are given in arithmetic average +/- SD. Weight (78.6 +/- 17.9; 75.7 +/- 17.8; 76.8 +/- 19.1 kg), Dauly insulin dose (65.4 +/- 15.1; 54.4 +/- 11.2; 54.8 +/- 9.3 IU), HbA1c (8.8 +/- 1.8; 8.2 +/- 1.1; 10.1 +/- 2.8%). Utilisation of glucose (3.5 +/- 1.6; 4.2 +/- 1.7; 4.4 +/- 1.8 mg/kg/min), triglycerides (1.2 +/- 0.5; 1.1 +/- 0.4; 1.3 +/- 0.7 mmol/l), cholesterol (5.1 +/ 0.7; 4.9 +/- 0.7; 5.2 +/- 0.8 mmol/l). CONCLUSIONS: The combination of metformin and the intensive insulin therapy in type 1 diabetic patients led, in contrast to placebo, to the significant reduction in weight (p < 0.001), to the reduction in insulin requirements (p < 0.05), to the improved control of glycaemia (p < 0.01) and to the decrease of FFA during clamp (p < 0.01). PMID- 11411060 TI - [Effect of coenzyme Q10 in patients with kidney diseases]. AB - BACKGROUND: Coenzyme Q10 belongs to important antioxidants and it has a key role in the synthesis of adenosinetriphosphate. Its beneficial effect was proved in several diseases, e.g. in mitochondrial encephalopathy, mitochondrial myopathy, mitochondrial cardiomyopathy. MATERIAL AND METHODS: All 15 patients of the studied group (5 with tubulopathy and 10 with chronic tubulointersticial nephritis) received antioxidative therapy for three months (E vitamin, C vitamin, riboflavin) and for the last two months coenzyme Q10 was added. Renal functions, spectrum of lipids, parameters of lipid peroxidation (malondialdehyde), levels of alpha-tocopherol, beta-carotene, coenzyme Q10. RESULTS: Before the substitutive antioxidative treatment, coenzyme Q10 levels reached in blood 0.11 +/- 0.03 mumol/l and 0.15 +/- 0.04 mumol/l in plasma. These values were well below the reference range (rr) is 0.4 +/- 1.0 mumol/l). After the substitution coenzyme Q10 levels significantly increased (p < 0.001) to the values of 1.66 +/- 0.16 mumol/l in blood and to 1.78 +/- 0.27 mumol/l in plasma. Plasma levels of beta-carotene increased from the markedly subnormal values 0.25 +/- 0.07 mumol/l (rr > 0.8 mumol/l) to 0.56 +/- 0.02 mumol/l (no statistical difference). Plasma levels of alpha-tocopherol remained within the reference range 32.15 +/- 4.73 mumol/l (rr 15-30 mumol/l) and they increased up to the plasma level of 44.83 +/- 5.82 mumol/l during the period of testing. Malondialdehyde levels did not significantly change within the testing period. No changes in renal functions and parameters of lipid metabolism were described. Patients well tolerated the treatment and no adverse effects were seen during the period of observation. CONCLUSIONS: Our results ascertained that levels of antioxidant CoQ10 were lower in patients with nephropathy who underwent conservative treatment with peroral substation. Such deficit can be amended by CoQ10 administration, which could be therefore taken as complementary treatment of nephrology. PMID- 11411061 TI - Computerized incident reporting: an example of healthcare IT for the 21st century. PMID- 11411062 TI - Analysis of hospital mortality for continuous improvement. PMID- 11411063 TI - Network liability: a new frontier for healthcare risk management. PMID- 11411064 TI - Incident reports may be discoverable in Nebraska. Nebraska ex rel. AMISUB v. Buckley, 260 Neb. 596 (2000). PMID- 11411065 TI - Hysterical fear of snakes not a disability under the ADA. Anderson v. North Dakota State Hospital 232 F. 3d 634 (8th Cir. 2000). PMID- 11411066 TI - Physician will not be liable for failing to disclose financial conflict of interest. Neade v. Portes, 193 Ill. 2d 433, 739 N.E. 2d 496 (Ill. 2000). PMID- 11411067 TI - State insurance fund is not entitled to privileged information in Wisconsin. Wisconsin Patients Compensation Fund v. Physicians Insurance Company of Wisconsin, 239 Wis. 2d 360 620 N.W. 2d 457 (Wis. 2000). PMID- 11411068 TI - An action for negligent credentialing is a professional liability action in Indiana. Winona Memorial Hospital v. Kuester 737 N.E. 2d 824 (Ct App. In. 2000). PMID- 11411069 TI - Hospital may be liable for actions of nurse who molested child. E.P. v. McFadden, - So. 2d, 2000 WL 303063, No. 298 (Ct. Civ. App. Ala. March 24, 2000). PMID- 11411070 TI - Addressing antimicrobial resistance threats. PMID- 11411071 TI - Endodontic problems and failures: how to anticipate, evaluate and prevent them. PMID- 11411072 TI - Cholesterol as an aetiological agent in endodontic failures--a review. AB - Presence of cholesterol clefts in apical periodontitis lesions is a common histopathological observation. However, a potential aetiological association of cholesterol crystals to non-resolving apical periodontitis lesions after endodontic treatment has been suggested only recently. This article is an attempt to explain the biological basis for the inability of body cells to eliminate the local accumulation of cholesterol crystals, and to consolidate available clinical and experimental data in support of the view that massive accumulation of cholesterol crystals in inflamed periapical tissues can interfere with the periapical healing after conventional root canal treatment. As the irritating cholesterol crystals and certain other aetiological agents of failed endodontic treatment exist outside the root canal system, it is concluded that re-treatment alone, of such cases, is unlikely to resolve the factors that sustain the lesion. Apical surgery is indicated for successful management of such cases. PMID- 11411073 TI - Clinical update--the teeth and the maxillary sinus: the mutual impact of clinical procedures, disease conditions and their treatment implications. Part 1. The differential diagnosis of tooth sinus pain--the dentist's view. AB - The close relationship of the teeth and the maxillary sinus requires both communication and cooperation between the dentist and the ENT specialist to ensure the best outcome for the patient. PMID- 11411074 TI - Clinical update--the teeth and the maxillary sinus: the mutual impact of clinical procedures, disease conditions and their treatment implications. Part 2. Odontogenic sinus disease and elective clinical procedures involving the maxillary antrum: diagnosis and management. AB - Although odontogenic sinusitis is a rare entity when compared to sinus disease of rhinogenic origin, it is extremely important to identify a dental aetiology when it occurs. The offending tooth or teeth would thus require endodontic treatment or extraction, and the sinus disease carefully assessed and appropriately managed. Aetiology and presentation of oro-antral fistulae are also discussed and guidelines for the management of this complication are recommended. Certain lesions such as cysts and tumours may involve the jaws and hence the maxillary antrum; some of these, such as a radicular cyst are quite common, but the rarer ones are included for completeness. Surgical techniques are continuously evolving to optimise form and function of the jaws, and when applied to the maxilla there may be some impact on nasal and sinus function. The advent, and now proven success, of osseointegrated jaw implants have brought with them innovations and refinements of bone grafting techniques, and more recently distraction osteogenesis for augmentation. Maxillary osteotomies for surgical orthodontics, and to facilitate prosthodontic treatment are briefly mentioned, as most of these inevitably involve the antrum and/or nose. This paper discusses, in summary form, important aspects of clinical dental practice which may involve the maxillary antrum. It is thus a broad overview of certain pathologic conditions and elective surgical procedures which have relevance to both medical and dental practitioners. PMID- 11411075 TI - Clinical update--the teeth and the maxillary sinus: the mutual impact of clinical procedures, disease conditions and their treatment implications. Part 3. But doctor, my ear hurts. AB - Temporomandibular disorders (TMD) can present as an array of symptoms that can mimic both odontogenic and ear, nose, and throat (ENT) disorders. Dentists need to be alert to the possibility of primary ENT disorders in patients who present with TMD symptoms, as should ENT physicians be familiar with the clinical presentation of TMD, especially in those patients who present with ENT symptoms but no sign of ENT disease. PMID- 11411076 TI - Clinical update--the teeth and the maxillary sinus: the mutual impact of clinical procedures, disease conditions and their treatment implications. Part 4. Pathogenesis and investigation of sinusitis. PMID- 11411077 TI - Above all do no harm. PMID- 11411078 TI - Clinical update--the teeth and the maxillary sinus: the mutual impact of clinical procedures, disease conditions and their treatment implications. Part 5. Medical and surgical management of sinusitis. PMID- 11411079 TI - ASOMAT revisited: conference on bio-compatible dentistry--September 1998. PMID- 11411080 TI - The problem of dentoalveolar ankylosis and subsequent replacement resorption in the growing patient. AB - Dentoalveolar ankylosis is a serious complication following injuries of the periodontal membrane in severe dental trauma such as avulsion and intrusion. The condition is a fusion of the alveolar bone with the tooth. The consequences of this condition are progressive resorption of the root with replacement by bone (replacement resorption) and arrested growth of the alveolar process in the growing patient. This article will present an overview of dentoalveolar ankylosis and replacement resorption and its problems, treatment options and prognosis, especially focusing on the growing patient. PMID- 11411081 TI - Prevention/treatment of resorption complications after trauma to the attachment apparatus. PMID- 11411082 TI - Watch this space: the missing anterior tooth--orthodontic considerations. PMID- 11411083 TI - Investigations on a cell culture medium for storage and transportation of avulsed teeth. AB - Non-physiologic storage of avulsed teeth leads to a high incidence of root resorption, resulting in poor prognosis. This study investigated the suitability of specially composed cell culture media for storage of extracted teeth for up to 48 hours. Autoradiographic investigations revealed that the proliferative activity of periodontal ligament (PDL) cells of teeth stored in cell culture medium for up to 48 hours increased with storage time. Studies on proliferation of PDL cells after storage of teeth in different media for up to 24 hours demonstrated that the proliferative activity is dependent on the composition of the medium. Immunohistochemical investigations with markers for cell proliferation revealed that pulp cells of extracted immature teeth show numerous proliferations after storage for up to 24 hours in a special cell culture medium but few proliferations after storage in Hanks Balanced Salt Solution (HBSS). The investigations indicate that a special cell culture medium can preserve cell viability of PDL cells adhering to extracted teeth for at least 48 hours. The in vitro results are confirmed by a case presented: After storage of two upper central incisors for 36 hours in the cell culture medium the teeth could be successfully reimplanted after extraoral insertion of titanium posts into the root canal (auto-alloplastic reimplantation). Clinical and radiological follow-up examinations for 12 months revealed normal periodontal healing. PMID- 11411084 TI - Inappropriate treatment of traumatic dental injuries. AB - Traumatic dental injuries are emergencies that must be treated expediently and efficiently to reduce pain and to restore function and appearance. With an increase in the incidence of traumatic dental injuries in our community (I) it is essential that the dental practitioner has "up-to-date" knowledge of dental trauma. The peak incidences of injury are 2-4 years and 8-10 years of age, with statistics revealing 30% of children suffer trauma to the primary dentition, and 22% of children suffer trauma to the permanent dentition by the age of 14 (I). The male to female ratio is 2:1. Aside from the emergency treatment and clinical decisions that must be made at the time of injury there is a need for long-term follow-up because of the high incidences of complications (2, 3). The factors that will influence the extent of injury will be energy impact, the direction of the impacting object, its shape and its resilience (4). Recent articles have raised concerns about inappropriate treatment for traumatic dental injuries (5, 6). This report will look at one such case. PMID- 11411085 TI - Invasive cervical resorption following trauma. AB - Invasive cervical resorption is an insidious and often aggressively destructive form of external root resorption which may occur as a late complication following dental trauma particularly where it involves damage to cementum and supporting tissues. While this resorption may be evident clinically as a pink coronal discolouration, later with cavitation of the enamel, often there are no obvious external signs and the condition is only detected radiographically. It is characterised by the invasion of the cervical region of the root by fibrovascular tissue which progressively resorbs dentine, enamel and cementum. The dental pulp remains protected by an intact layer of dentine and predentine until late in the process. Ectopic calcifications can be observed in advanced lesions both within the invading fibrous tissue and deposited directly onto the resorbed dentine surface. The aetiology of invasive cervical resorption is unknown but trauma has been documented as a potential predisposing factor. A recent study by the author of 222 patients with a total of 257 teeth which displayed varying degrees of invasive cervical resorption showed that trauma alone was a potential predisposing sole factor in 14% of patients and 15.1% of teeth. Trauma in combination with bleaching, orthodontics or delayed eruption was found in an additional 11.2% of patients or 10.6% of teeth and of these a combination of trauma and bleaching occurred in a relatively high proportion of 7.7% of patients or 7.4% of teeth. This study also revealed that of other potential predisposing factors orthodontics was the most common sole factor constituting 21.2% of patients and 24.1% of teeth examined. Successful treatment of invasive cervical resorption is dependent on the extent of the resorptive process. Teeth with invasive cervical resorption have been divided into four classes. Whilst several treatment modalities are possible, a clinical evaluation of the treatment of this condition by the topical application of a 90% aqueous solution of trichloracetic acid, curettage, endodontic therapy where necessary and restoration with a glass ionomer cement has been evaluated on 94 patients with a total of 101 teeth with a minimum follow-up period of three years. Results indicate a satisfactory treatment outcome can be anticipated in Class 1, 2 and 3 cases. In Class 4 resorption no treatment or alternative therapy is recommended. Diagnosis of lesions at an early stage of development is highly desirable and therefore the patients who have a potential for the development of this condition by virtue of a history such as trauma should be monitored radiographically at intervals throughout life. PMID- 11411086 TI - Increasing mouthguard usage. A pilot campaign for junior basketball and Rugby. PMID- 11411087 TI - How could it go so quickly? PMID- 11411088 TI - 10th International Conference on Dental Traumatology, Friday March 19-Sunday March 21, 1999. PMID- 11411089 TI - [Immunological mechanisms of acquired aplastic anemia]. PMID- 11411090 TI - [Autoimmune hepatitis associated with mixed connective tissue disease: report of a case and a review of the literature]. AB - A case is reported of a 29 year old female who had autoimmune hepatitis associated with mixed connective tissue disease (MCTD). The patient developed MCTD at the age of 19, and was treated with prednisolone. Liver dysfunction developed 7 years later, which exacerbated shortly after the patient suffered intrauterine fetal death during the second trimester of pregnancy. Laboratory data showed negative anti-hepatitis C antibody and hepatitis B antigen, but positive anti-smooth muscle antibody. A liver biopsy showed chronic active hepatitis. Referring to the criteria we diagnosed her as having autoimmune hepatitis. Although hepatomegaly is sometimes observed in MCTD patients, only 5 cases of autoimmune hepatitis associated with MCTD have been reported in the past. In our case, it is of note that autoimmune hepatitis developed while symptoms of MCTD were in remission, and that autoimmune hepatitis exacerbated with the emergence of anti-smooth muscle antibody following the termination of pregnancy. PMID- 11411091 TI - [Association of idiopathic thrombocytopenic purpura and type 1 diabetes mellitus in a patient with sarcoidosis]. AB - Occurrence of autoimmune diseases with sarcoidosis is well known. However, a case in which more than one of these diseases coexist with sarcoidosis is rare. We present a young man with suspected sarcoidosis, complicated by idiopathic thrombocytopenic purpura (ITP) and type 1 A diabetes mellitus (DM). A 21-year-old man was admitted to our hospital because of thrombocytopenia, hyperglycemia, and bilateral hilar lymphadenopathy (BHL). Although a histological proof could not be obtained, the patient was considered to have sarcoidosis because 67-gallium scintigraphy disclosed "Lambda" and "Panda" signs which are highly specific for sarcoidosis. Type 1 A DM was also diagnosed as the patient had antiglutamic acid decarboxylase antibodies. The patient disclosed no hepatosplenomegaly or no lymphadenopathy and diagnosis of ITP was confirmed by bone marrow examination. High dose steroid was started as the thrombocytopenia progressed. The platelet number increased satisfactorily and shrinkage of BHL was also observed with the therapy. PMID- 11411092 TI - [Antimicrobial prophylaxis in oral and maxillofacial surgery]. PMID- 11411093 TI - Effects of water deprivation on neurokinin B production by the arginine vasopressin neurons of hypothalamic paraventricular and supraoptic nuclei. AB - In order to clarify the role of neurokinin B (NKB), the dynamic changes in NKB expression and synthesis following water deprivation were examined in the arginine-vasopressin (AVP) neurons of hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei. In intact rats, NKB and AVP showed almost the same high level of immunohistochemical reactivity in the magnocellular neurons of the PVN and SON, as well as in the varicose fibers in the median eminence (ME). In contrast, NKB precursor peptide (NKBp) immunoreactivity in the SON and PVN were relatively weak. Five days after water deprivation, AVP and NKB immunoreactivity decreased drastically, while NKBp-immunoreactivity increased in both the PVN and SON magnocellular neurons. Reverse transcription-polymerase chain reaction analysis of control animals revealed high levels of AVP mRNA and substantial amounts of NKB mRNA in the SON. This was contrast to the relatively low levels of AVP mRNA and undetectable levels of NKB mRNA in the PVN. After five days of water deprivation, AVP mRNA in the PVN and NKB mRNA in both the PVN and the SON increased considerably. These results indicate that synthesis and release of NKB, which colocalizes to AVP neurons, are enhanced by water deprivation in the same manner as AVP in the PVN and SON. Thus, NKB seems to be involved in the central control of body fluid levels. The results also suggest that the production rate of NKB under normal conditions in SON dominant. PMID- 11411094 TI - High-dose vitamin C therapy for inclusion body myositis. AB - OBJECTIVES: The efficiency of high-dose vitamin C therapy for inclusion body myositis (IBM) was assessed. SUBJECTS & METHODS: The subjects were five patients with IBM confirmed pathologically. After the intravenous administration of 40 mg/kg vitamin C five times/week for four weeks, muscle weakness was found to improve in three cases. The average muscle score improved from 8.1 to 8.8, from 7.0 to 8.1 and from 6.2 to 6.8. Magnetic resonance imaging (MRI) demonstrated a reduction in the size of T2 high lesions and gadolinium enhancement in the thigh muscles in one case. Based on our findings, high-dose vitamin C therapy is considered to be effective in some cases of IBM. PMID- 11411095 TI - [Clinicopathological study of influential factors on prognosis in patients with renal cell carcinoma]. AB - Between January 1986 and August 1996, 120 patients were diagnosed with renal cell carcinoma (RCC) at Kanagawa Cancer Center. In this study, we analyzed these patients in view of the clinicopathological factors, which affect the prognosis of these patients with univariate and multivariate analysis using Cox's proportional hazard model. Of these 120 patients, 40 patients (33%) died of RCC during this period. The cause specific survival rate was 65% after 5 years and 57% after 10 years. Univariate analysis showed some factors, performance status, symptoms, tumor size, histological grade, local invasion (pT), venous involvement (pV), lung metastasis, lymph-node metastasis, and bone metastasis, affected the prognosis. Furthermore, multivariate analysis using Cox's proportional hazard model showed that performance status, pT, pV, lung metastasis, lymphnode metastasis, and bone metastasis independently affected the prognosis. In summary, TMN factors and performance status importantly affect the prognosis of RCC patients. PMID- 11411096 TI - [A clinicopathological evaluation of prognostic factors of urothelial tumors of the renal pelvis and ureter]. AB - We investigated the clinicopathological features of 109 patients with urothelial tumors of the renal pelvis and ureter who underwent surgery at four institute from April, 1975 to September, 1997. The patients consisted of 71 males and 38 females, and the mean age was 66.8 years, ranging from 41 to 92 years. Mean followup period was 43 months. The prognostic significance of the pathological factors; pT, Grade, INF, tumor size, pL, pV and pN were evaluated. All these factors affected the survival rates significantly in univariate analysis. Multivariate analysis showed the most influential prognostic factors to be pT and pN. PMID- 11411097 TI - [Study of neoadjuvant chemotherapy for invasive bladder cancer with MEC (methotrexate, epirubicin, cisplatin) therapy]. AB - Eighteen patients with locally invasive bladder cancer were treated with 1 or 2 cycles of neoadjuvant chemotherapy consisting of methotrexate, epirubicin and cisplatin (MEC). All patients underwent radical cystectomy and pelvic lymph node dissection. Down-staging was observed in twelve (complete pathological response in 3 and partial pathological response in 9) patients (response rate were 67%). Four of the 18 patients died of disease and all of them had not achieved down staging. Multivariate analysis revealed nodal status to be the only independent predictor. With regard to side effects, gastrointestinal symptoms and myelo suppression were observed in almost all patients. Thrombocytopenia was observed in 13 patients (72%) including 7 patients who showed symptoms over grade 3. Gastro-intestinal symptoms and leukocytopenia disappeared with granisetron and granulocyte colony stimulating factor. There were no treatment-related deaths in this study. These results indicate that MEC therapy was safely performed and showed a high response rate in patients with locally invasive bladder cancer. PMID- 11411098 TI - [Clinical evaluation of urinary nuclear matrix protein 22 as a marker for bladder cancer]. AB - The purpose of this study is to evaluate the clinical usefulness of urinary nuclear matrix protein 22 (NMP22) as a marker for bladder cancer. We examined the positive rates of NMP22 test, urinary cytology and bladder tumor antigen (BTA) test, and compared the positive rate of NMP22 test with that in urinary cytology and BTA test. Urine samples were obtained from 50 patients with histologically confirmed bladder cancer before the treatment. The samples were examined by NMP22 test, urinary cytology and BTA test. In 50 patients with bladder cancer, the overall positive rate was 40% for NMP22 test, 40% for urinary cytology, and 16% for BTA test. A combination of NMP22 test and urinary cytology showed a significantly higher positive rate (54%) as compared to NMP22 test or urinary cytology alone. When NMP22 test and urinary cytology were compared for tumor size, number, shape, stage and grade, NMP22 test showed a significant higher positive rate than urinary cytology in grade 1 bladder cancer. In conclusion, although NMP22 test and urinary cytology gave a similar positive rate, a combination of NMP22 test and urinary cytology is more useful than the NMP22 test or urinary cytology alone for monitoring of bladder cancer. PMID- 11411099 TI - [Clinical efficacy of treatment with low-dose flutamide in maximum androgen blockade therapy]. AB - To prevent treatment withdrawal due to flutamide-induced liver dysfunction, we performed maximum androgen blockade (MAB) therapy by combining a luteinizing hormone-releasing hormone agonist or orchiectomy with low-dose flutamide (125 mg x 2/day) in patients with prostate cancer. In this study, the efficacy, adverse effects such as hepatotoxicity, and compliance were compared retrospectively between 35 patients who received low-dose flutamide therapy (1995-1999) and 27 patients who received flutamide at its ordinary dose (125 mg x 3/day). No significant difference was observed in the response rate (> or = PR) as determined from the prostate-specific antigen parameter (p = 0.6211) or the incidence of hepatotoxicity based on the aspartate aminotransferase and alanine aminotransferase levels. However, flutamide withdrawal due to liver dysfunction was less frequent in the low-dose group (2.9%) than in the ordinary dose group (18.5%) (p = 0.0386). MAB therapy using low-dose flutamide is expected to prevent the reduction in the compliance due to side effects and to improve the long-term prognosis in patients with prostate cancer, who are mostly elderly individuals. PMID- 11411100 TI - [A case of malignant pheochromocytoma associated with von Recklinghausen's disease]. AB - A 48-year-old woman suffering from chest and lumbar pain was referred to our clinic for treatment. She had a history of von Recklinghausen's disease with multiple cafe-au-lait spots and neurofibromatosis. Computed tomography demonstrated a large tumor in the left adrenal gland and a small lesion in the liver, with the levels of plasma and urinary catecholamines being elevated. 131I metaiodobenzylguanidine (MIBG) scintigraphy showed abnormal accumulations in the left adrenal tumor and multiple-bone lesions. A diagnosis of malignant pheochromocytoma with liver and bone metastases was made, and the patient received chemotherapy. Seven months after the diagnosis of malignant pheochromocytoma, she died of pulmonary edema due to disease progression. Autopsy revealed malignant pheochromocytoma with liver, lung, bone and lymph nodes metastases. We reviewed the literature on pheochromocytoma associated with von Recklinghausen's disease. To our knowledge, only 7 cases of malignant pheochromocytoma associated with von Recklinghausen's disease have been reported in Japan. PMID- 11411101 TI - [Von Hippel-Lindau disease associated with renal cell carcinoma and bilateral cystadenoma of the epididymis: a case report]. AB - We present a case report of von Hippel-Lindau disease associated with renal cell carcinoma and bilateral cystadenoma of the epididymis. A 26-year-old man appeared with painless tumors of the bilateral scrotal contents. Ultrasonography and other radiographic examinations including computed tomographic scan and dripinfusion pyelography showed multiocular tumors in the bilateral epididymis and a right renal tumor 3 cm in diameter. The tumors of the bilateral epididymis were surgically resected and of the right renal tumor enucleated. Histopathological examination revealed cystadenoma of the epididymis and renal cell carcinoma (clear cell carcinoma, G1, pT1a). He has not received adjuvant therapy, and is doing well with no evidence of metastatic disease 2 years after surgery. PMID- 11411102 TI - [Spontaneous renal rupture resulting from ureteral tumor left untreated for 6 years: a case report]. AB - A 57-year-old man, 6 years after discovery of a left ureteral tumor was admitted to our hospital complaining of severe left abdominal pain. With the diagnosis of acute abdomen, the patient was examined by computed tomography, which showed severe left hydronephrosis with renal rupture. Retrograde and antegrade pyelography showed the hydroureteronephrosis due to the tumor in the lower ureter, so left nephroureterectomy was performed. The ureteral tumor was diagnosed as transitional cell carcinoma, grade 3 and pT2. The present case is the 9th case of spontaneous renal rupture caused by renal pelvic and ureteral tumors in Japan. PMID- 11411103 TI - [A case of urothelial tumors which occurred simultaneously in bilateral upper urinary tracts and bladder presenting with anuria]. AB - A 72-year-old male was admitted with a chief complaint of anuria. Clinical examinations showed that he was in uremic state and had bilateral hydronephroses. An endoscopic examination revealed a left ureteral tumor and a bladder tumor. Left nephroureterectomy with partial cystectomy and transurethral resection of the bladder tumor were performed. Pathological examinations showed an invasive left renal pelvic tumor (pT3, G3), an invasive left ureteral tumor (pT4, G3), and a bladder tumor (pTis, G3). Following the operation, roentgenological and urinary cytological findings showed a right ureteral carcinoma. He died of multiple liver and bone metastases and local recurrence at 5 months postoperatively. PMID- 11411104 TI - [Small cell carcinoma of the urinary bladder with synchronous esophageal cancer and incidental lung cancer: a case report]. AB - We present a case of triple primary cancers occurring synchronously in the urinary bladder, esophagus, and incidentally in the lung. A 65-year-old man with a chief complaint of gross hematuria was admitted to our hospital. Cystoscopy, computed tomography (CT) and magnetic resonance imaging (MRI) revealed a non papillary broad-based bladder tumor. Histological diagnosis was transitional cell carcinoma of the urinary bladder and he underwent one course of neoadjuvant chemotherapy (M-VAC) with the preoperative diagnosis of T3bN0M0. After one course of chemotherapy, chest CT, lymph node biopsy and esophagoscopy revealed squamous cell carcinoma of the esophagus. He first underwent radiochemotherapy (total 70 Gy, CDDP 5 mg x 41, 5-FU 250 mg x 24) for esophageal cancer and achieved complete remission. Then, he underwent radiotherapy for a total of 60 Gy for bladder cancer. However, his general condition gradually became worse and he died from metastatic cancer. The autopsy proved that he died from multiple metastases of small cell carcinoma of the urinary bladder and incidentally squamous cell carcinoma of the lung was identified. PMID- 11411105 TI - [Transitional cell carcinoma of the bladder in a young patient: a case report]. AB - A 14-year-old male patient was admitted with the chief complaint of macroscopic hematuria. Abdominal ultrasonography demonstrated a tumor of the anterior wall of the bladder. Further, cystoscopic examination confirmed a papillary tumor. Transurethral resection of the bladder tumor was performed. Histopathology of the excised tumor showed transitional cell carcinoma (G1, pTa). Recurrence has not been observed for 9 months postoperatively. PMID- 11411106 TI - [Two cases of successful treatments with steroid for local and systemic hypersensitivity reaction following intravesical instillation of Bacillus Calmette-Guerin]. AB - We have found steroid pulse therapy to be effective and safe for local and systemic adverse reactions of BCG therapy. Two cases are reported. Case 1: A 57 year-old woman with initial recurrence of urinary bladder carcinoma was treated with transurethral resection. The histopathological findings were transitional cell carcinoma (TCC), G2 > G1, pT1a. To prevent a second recurrence, she was administered Bacillus Calmette-Guerin (BCG) instillation therapy: 80 mg of BCG, (Tokyo strain) suspended in 40 ml of normal saline, instilled into her bladder weekly. After the fifth week of instillation, she was found to have a cough, sputum, edema of the eyelids, congestion of palpebral conjunctive, severe pain on micturition and pollakisuria. Although she was administered antituberculus, antibiotics and antiallergic drugs, all sign and symptoms were aggravated. Blood, urine and sputum cultures remained negative for mycobacterium. She was later diagnosed as having hypersensitive reactions against BCG and treated with steroid pulse therapy. The signs and symptoms mentioned above were decreased immediately and disappeared after a week. Case 2: A 76-year-old man with initial recurrence of urinary bladder carcinoma was treated with transurethral resection. To prevent a second recurrence, he was instilled the BCG six (6) times. Although no adverse reaction was observed, urinary cytology remained positive (class V) and small papillary tumor was detected at the dome of the bladder. Transurethral biopsy was then performed. The histopathological findings showed TCC, G3, CIS on the dome of bladder. Then he was again administered the same BCG instillation therapy. After the fifth instillation, he complained of severe pain of micturition, pollakisuria and dysuria. Although he was administered antibiotics and antiinflammatory drugs, all signs and symptoms were aggravated. Urine culture remained negative for mycobacterium. He was diagnosed as having hypersensitive reactions against BCG and was treated with two times of steroid pulse therapy. The signs and symptoms mentioned above were decreased immediately and disappeared after the second steroid pulse therapy. PMID- 11411107 TI - [A case of septic shock due to ileal conduit obstruction]. AB - A 70-year-old man with bladder cancer received a total cystectomy and an ileal conduit 64 months before he visited our hospital with complaints of lower abdominal pain, shaking and chilliness. Bilateral hydronephrosis due to an ileal conduit obstruction were observed on the ultrasonography. Bacterial culture from blood and urine samplings revealed E. coli. Under the diagnosis of urosepsis, the administration of anti-biotics and bilateral percutaneous nephrostomy were performed. However, he suffered from septic shock and disseminated intravascular coagulation (DIC). Therefore, the treatments for DIC were done, and they were effective. The obstruction of the ileal conduit was cured spontaneously. No recurrence or metastases were found on ultrasonography and computed tomographic scan for 12 months after these treatments. PMID- 11411108 TI - [A case of infravesical obstruction caused by prostatic stones as diagnosed by transrectal ultrasonography at voiding]. AB - A 69-year-old patient, who had been treated using alpha 1-blocker for benign prostatic hyperplasia (BPH) at another clinic, visited our clinic due to persistent difficulty in urination. Total International Prostate Symptom Score (IPSS) was 32 points and quality of life (QOL) index was 5. Uroflowmetry demonstrated maximum urinary flow rate and average urinary flow rate to be 9.2 ml/sec and 5.1 ml/sec, respectively, with 3 ml of residual urine volume. Transrectal ultrasonography (TRUS) revealed prostatic stones but not BPH. Retrograde urethrography demonstrated nothing abnormal other than prostatic stones. TRUS at voiding phase using linear probe (voiding TRUS) revealed poor opening of the urethra surrounded by prostatic stones. As a result, the cause of urinary disturbance was diagnosed to be due to urethral obstruction caused by prostatic stones, and transurethral resection of prostatic tissue with stones was performed. Postoperatively, IPSS decreased to 10 points and QOL index to 2. Maximum urinary flow rate also improved to 18.1 ml/sec and mean urinary flow to 8.4 ml/sec. Thus, voiding TRUS is likely the best urodynamic test for clinical use in determining the etiology of obstruction at posterior urethra. PMID- 11411109 TI - [Sclerosing Sertoli cell tumor of the testis: a case report]. AB - We report here a case of sclerosing Sertoli cell tumor of the testis. A 21-year old male, who complained of right testicular pain, visited a Jikei University affiliated hospital on May 30th, 1999. A small nodule with a diameter of 6 to 7 mm was palpable on the central surface of the right testis. No tumor markers for testicular cancer, such as hCG-beta and alfa-fetoprotein, were elevated. However, ultrasound revealed a hypoechoic mass with increased blood flow. Therefore, we performed right high orchiectomy under the diagnosis of right testicular cancer. Pathological diagnosis of this tumor was sclerosing Sertoli cell tumor. Neither recurrence nor metastasis has been found for 12 months postoperatively. PMID- 11411110 TI - [Definition, concept and pathophysiology of pulmonary hypertension]. AB - Intravenous prostacyclin infusion has been introduced as breakthrough to the treatment of primary pulmonary hypertension(PPH) followed thus far inevitably by calamitous outcome and has recently promised a favorable outlook. Pulmonary hypertension (PH) is defined as pulmonary artery mean pressure exceeds 25 mmHg at rest in a lying position. The concept of PH was discussed historically down to date and has been usually used for the patient with sustained PH in vascular origin including PPH. Pathophysiology relating to the cause of PH has been focused on unexplained PH which is PH due to collagen, hepatoportal, congenital cardiac diseases, HIV infection and anorexigenics. Various intrinsic chemical mediators such as TXA2, ET-1, serotonin, eNOS, K channel and their antagonists give rise to PH. Genetics of PH was discussed around Gene PPH-1 disclosed by Genome Project. However, further attention should be paid for enormous pulmonary vascular reserve in normals, because PH occurs only when this reserve must be reduced to a critical level before medial muscular hypertrophy develops. PMID- 11411111 TI - [Epidemiology of primary pulmonary hypertension in Japan]. AB - Primary pulmonary hypertension(PPH) is a progressive disease of unknown etiology with poor prognosis. Recent epidemiological studies in Japan have shown that there are 230 patients (95% confidence interval: 200-260) with mean age of 41.8 +/- 18.5 years old, female to male ratio of 1.72: 1 and their median survival time was 69.0 months. At a later time, the clinically effective modalities such as lung transplantation and prostaglandin I2 for PPH were available in Japan. In 1999, the Ministry of Health and Welfare of Japan approved PPH as a objective disease of therapeutic study. Once patient is approved as PPH based on the personal clinical inquiry, Doctor's fee will be paid at the public government. This system will promote earlier and more accurate diagnosis in PPH patients and will clarify more reliable clinico-epidemiological figure of PPH in a whole territory of Japan. PMID- 11411112 TI - [Coagulation and fibrinolytic abnormality related to endothelial injury in pulmonary arterial hypertension]. AB - It remains unclear whether a coagulation abnormality plays an important role in the clinical course in pulmonary arterial hypertension as well as vasoconstriction and vascular-wall remodeling, although several studies of pathological specimens and clinical studies demonstrating that anticoagulation prolongs life have suggested the importance of thrombosis. Several parameters for hemostasis related to endothelial injury are expected to reflect abnormal coagulation and fibrinolytic systems in pulmonary hypertension. Recently, we demonstrated that plasma level of soluble P-selectin was increased and that of thrombomodulin (TM) was decreased in patients with pulmonary arterial hypertension. The abnormal values of P-selectin and TM were improved by continuous prostacyclin infusion. Altered hemostasis may be important as a pathogenesis in pulmonary hypertension and adequate anti-coagulant therapy is expected. PMID- 11411113 TI - [Nitric oxide and endothelins in pulmonary hypertension]. AB - For recent decades, the medical treatment of primary pulmonary hypertension(PH) have shown an improved outcome as a bridge to lung transplantation. Both nitric oxide (NO) and endothelin(ET) have reported as a major vasoactive mediators in initiating PH. NO is a potent vasodilator released from endothelial cells to adjacent smooth muscle cells in vascular wall. Inhalation of NO has exerted dramatic effects with minimum complication for PH patients. ET-1, after binding to ETA receptor, promotes a strong vasoconstriction, cell growth and platelet aggregation. ETB receptor promotes the clearance of circulating ET-1 in the lung, and stimulates the release of NO and PGI2. Therapeutic trials of ETA receptor antagonist and NO donors for PH may be promising because of their direct action against vasoconstriction. PMID- 11411114 TI - [Alternation of regulation system for pulmonary arterial tone in pulmonary hypertension]. AB - In pulmonary artery smooth muscle cells(PASMC), the Ca-influx required for myogenic tone is mainly supplied through the L-type Ca channels, which sustained open around resting membrane potential. Therefore, the voltage-gated K channels, which determine membrane potential in PASMC, are critical for regulation of the Ca-influx and myogenic tone. The voltage-gated K channels are inhibited in hypoxic condition. The inhibition may be related to the hypoxic pulmonary vasoconstriction and the chronic hypoxia-induced pulmonary hypertension. In subjects with primary pulmonary hypertension, the voltage-gated K channels are also inhibited. On the other hand, the Ca-activated K channels of PASMC are also inhibited in chronic exposure with pulmonary hypertension. The reduction of the K channels in pulmonary hypertension may enhance proliferation of PASMC. PMID- 11411115 TI - [Fawn-Hooded Rat; an animal model of development of pulmonary hypertension]. AB - The Fawn-Hooded Rat (FHR) spontaneously develops pulmonary hypertension (PH) at sea level, and an increased severity of this disease is observed upon exposure to mild hypoxia. A recent report suggested that lung hypoplasia with decreased alveolarization and altered vascular growth led by the decreased activity of endothelial nitric oxide synthase may contribute to the development of PH in the FHR. Exposure to mild hypoxia (P1O2 = 120 mmHg) leads to severe PH in FHR but not in Tester Moriyama rat, a strain that has a serotonin platelet storage-pool deficiency (PSPD) similar to that of the FHR. A serotonin PSPD does not appear to predispose FHR to PH. Endothelin-1 (ET-1) mRNA and peptide levels are increased in the hypertensive lungs of mildly hypoxic FHR. ET-1 may at least partly contribute to the development of PH in this strain. PMID- 11411116 TI - [Monocrotaline-induced pulmonary hypertension in animals]. AB - Monocrotaline (MCT) is a pyrrolizidine alkaloid extracted from seeds of Crotalaria spectabilis that has been used to induce structural changes in the pulmonary arteries associated with progressive pulmonary hypertension in rodents. Recent studies have demonstrated that several mediators such as elastase, vasoconstrictors (Endotheline-1), vasodilators (Nitric oxide, PGI2) and cytokines (IL-1, MCP-1) play important roles in MCT-induced pulmonary hypertension, and that the changes in pulmonary hemodynamics are required for neointimal formation in pulmonary arteries. We have recently provided the evidence that macrophage migration inhibitory factor (MIF) is also involved in this model. The MCT-induced pulmonary hypertension in animals is useful as an experimental model for understanding the pathophysiology of primary pulmonary hypertension. PMID- 11411117 TI - [Pulmonary hypertension in endothelial NO synthase knockout mice]. AB - Inhalation of nitric oxide (NO) effectively reduces pulmonary arterial pressure in patients with pulmonary hypertension. However, it is controversial as to the role of endothelium-derived NO in pulmonary hypertension. Disruption of endothelial NO synthase (eNOS) gene causes systemic hypertension and diminishes endothelium-dependent vasodilatation. Even at normoxia eNOS knockout mice showed slightly increased right ventricular systolic pressure, but normal right ventricular weight. Hypoxia increased right ventricular systolic pressure, hematocrit, right ventricular weight in a concentration-dependent manner. The hypoxic condition induced pulmonary vascular remodeling; increases in the ratio of fully muscular vessels. Thus, pulmonary hypertension in eNOS knockout mice may be due to exaggerated vasoconstriction in response to hypoxia. In contrast, eNOS gene transduction reduces pulmonary vascular resistance. PMID- 11411118 TI - [Angiography of pulmonary hypertension]. AB - In the present report, we reviewed articles on pulmonary angiography for patients with pulmonary hypertension. The use of a non-ionic, low-osmolarity agent and improvements in catheters reduced periprocedual complications, but attention should be paid to the pre-existing poorer physical state. Digital pulmonary angiography has similar diagnostic power to conventional angiography. Wedged pulmonary angiography could demonstrate distal lesions of the pulmonary artery which couldn't be detected by conventional pulmonary angiography. Typical pulmonary angiograms from pulmonary hypertension due to pulmonary embolism, primary pulmonary hypertension, collagen diseases, Takayasu's arteritis and porto pulmonary hypertension were shown. Pulmonary angiography is relatively less valuable than before because of advances in other diagnostic imaging tools, but still remains the gold standard. PMID- 11411119 TI - [The role of cardiac catheterization for diagnosis and treatment of pulmonary hypertension]. AB - The role of the cardiac catheterization for diagnosis and treatment of pulmonary hypertension (PH) is very important. When mean pulmonary artery pressure increased more than 25 mmHg, then PH is defined. But this is measured accurately only by the catheterization. And we can discriminate the etiology of PH clearly by pulmonary capillary wedge pressure (Ppcw) or intra-cardiac shunt (L to R) by blood oxygen saturation step-up, and both parameters are obtained by this method. The etiology of PH is diagnosed as left sided heart failure, if Ppcw is increased more than 13 mmHg. PH is produced by congenital heart disease (ASD, VSD, PDA etc.), when the oxygen saturation step-up is recognized. And PH is induced by any pulmonary disease or pulmonary thrombo-embolism or collagen disease or liver cirrhosis or PPH, if Ppcw is normal and no oxygen step-up is recognized. PMID- 11411120 TI - [PH score--a new scoring system for pulmonary hypertension with chronic lung disease]. AB - Pulmonary hypertension is the most serious complication with the infants of chronic lung disease and pulmonary hypertension. There were no evaluating methods available for outpatients. We've made the scoring system from seven parameters of echocardiography which consist of RSTI, AT/ET, LV (S/L), RVaw (d), RVaw (s), P/A, T/M. The points of total PH score are correlated with estimated pulmonary arterial pressure from the maximum velocity of tricuspid regurgitation. The infants of points less than 2 are considered no PH, points 2 to 4 are mild PH, points 5 to 8 are moderate PH, points more than 8 are severe hypertension. The system has revealed useful for management of patients with home oxygen therapy. PMID- 11411121 TI - [CT and MRI findings of pulmonary hypertension]. AB - Although computed tomography (CT) and magnetic resonance imaging (MRI) can not measure the pulmonary arterial pressure, those can depict the morphological changes due to pulmonary hypertension, which are dilatation of main and central pulmonary artery, right ventricular hypertrophy and dilatation of right ventricle, right atrium, vena cava and coronary sinus. Right ventricular volume, mass and ejection fraction are calculated quantitatively by MRI using Simpson method. Thromboembolism can be detected by enhanced CT. Information about pulmonary blood flow and tricuspid regurgitation are given by MRI. Three dimensional MR angiography depicts the pulmonary artery as a whole. CT and MRI can detect pulmonary and congenital heart disease, the cause of pulmonary hypertension. CT and MRI are useful complementary method for evaluating pulmonary hypertension. PMID- 11411122 TI - [Value of nuclear medicine in pulmonary hypertension]. AB - Clinical applications of nuclear medicine in pulmonary hypertension (PH) include: 1) Differentiation of primary from chronic thromboembolic form of PH by mismatched ventilation/perfusion defects, 2) Quantification of right ventricular ejection fraction to estimate cardiac functional derangement, 3) Estimation of right ventricular hypertrophy or pressure overload by increased accumulation of radioactive myocardial perfusion agents, and 4) Estimation of myocardial sympathetic nervous function or fatty acid metabolism. Gravity dependent shift of pulmonary perfusion detected by perfusion scintigraphy has also been utilized for estimating severity of PH. Positron emission tomography (PET) has visualized distribution of pulmonary adrenergic receptors and angiotensin converting enzyme. PET, in addition to F-18 FDG lung tumor PET, should be watched for because it may visualize and quantify other bioactive substances and receptors in the lung. PMID- 11411123 TI - [Indication for lung biopsy and usefulness of lung biopsy]. AB - The criteria of open lung biopsy for operative indication were determined on the basis of hemodynamics in each congenital cardiac anomalies with pulmonary hypertension. More than 10.6, 8 and 8 units. m2 of pulmonary vascular resistance (PVR) were considered as indication for open lung biopsy in the patients with complete transposition of the great arteries, complete atrioventricular canal defect and atrial septal defect respectively. Lung biopsy should be done in patients with ventricular septal defect when PVR is more than 8 units. m2 and if it is greater than 4 with the oxygen inhalation test or 6 with the Tolazoline test. The usefulness of lung biopsy was also emphasized on the basis of the postoperative result in which lung biopsy diagnosis has been done. PMID- 11411124 TI - [Inhaled nitric oxide therapy]. AB - Inhaled nitric oxide (INO) therapy is becoming an indispensable measure in some critically ill patients with pulmonary hypertension. Most importantly, it has been shown that INO significantly reduces the necessity for extracorporeal lung assist in newborns with hypoxemic respiratory failure and pulmonary hypertension. A large number of basic and clinical studies are making great steps in delineating its physiology, techniques, side effects, and clinical efficacy. This article reviews the current knowledge of INO therapy including a novel device of nitric oxide production system. PMID- 11411125 TI - [Current status and future prospect of prostacyclin therapy for pulmonary hypertension--intravenous, subcutaneous, inhaled and oral PGI2 derivatives]. AB - Prostacyclin (PGI2) is a potent and a promising vasodilative agent in the treatment of pulmonary hypertension (PH). Epoprostenol (Flolan), an intravenous PGI2, has been used for PPH, and exerts dramatic effects such as reducing pulmonary vascular resistance and increasing cardiac output leading to an improvement of exercise tolerance. However, continuous i.v. infusion accompanies infection, thrombosis, occlusion, battery problems and variable side effects of PGI2 itself. Uniprost, administered subcutaneously, has a rather long half-life. It also needs an ambulatory infusion pump system. Beraprost, an oral PGI2 analogue, is also effective in mild PH patients. It can be absorbed easily and administered tid, or qid fashion. Inhaled iloprost may be an alternative option, but both iloprost and beraprost require frequent administrations because of their short half-life. PMID- 11411126 TI - [Endothelin receptor antagonist, phosphodiesterase inhibitor, thromboxane inhibitor]. AB - According to the advanced comprehension of pathophysiology of primary pulmonary hypertension (PPH), a therapeutical approach to PPH has changed recently. One of the breakthrough to the treatment of PPH is application of prostacyclin. It has been revealed that intravenous administration of prostacyclin has improved the prognosis and patient's quality of life. Another development of endothelin receptor antagonists and phosphodiesterase inhibitors have provided a novel pulmonary-specific effect. An endothelin receptor antagonist has a great inhibitory effect against pulmonary vasculature remodeling. In this regard, this regard, this receptor antagonist has superior effect to other medicines. Furthermore, a phosphodiesterase inhibitor shows a great decreasing effect on pulmonary hypertension with less effect on systemic blood pressure. These drugs will provide a great potential to the treatment of pulmonary hypertension. PMID- 11411127 TI - [Current status and future of lung transplantation]. AB - Lung transplantation has been performed successfully since 1983 in patients with various end-stage lung diseases including primary pulmonary hypertension. More than 10,000 lung transplants have been reported in The Registry of the International Society for Heart and Lung Transplantation. In contrast, a transplant law became effective in Japan only recently and 11 lung transplants have been performed with excellent results. We performed the first successful living-donor lobar lung transplantation for a 19-y-o-f with primary pulmonary hypertension on January 5, 2001 using her father's right lower lobe and her mother's left lower lobe. When the patients with primary pulmonary hypertension do not respond to prostacyclin therapy, lung transplantation is a workable option. PMID- 11411128 TI - [Gene therapy for pulmonary hypertension]. AB - Pulmonary hypertension is a life-threatening disease characterized by progressive vascular remodeling, ultimately producing right ventricular failure. Recently, continuous intravenous infusion of prostacyclin has become recognized as a therapeutic breakthrough that can improve hemodynamics and surgical in patients with pulmonary hypertension. Nevertheless, gene therapy is also expected as a valuable therapeutic approach in pulmonary hypertension for the reason that transgene expression is located mainly in the lung. Previous study demonstrated that intratracheal gene transfer of endothelial derived nitric oxide synthase, prostacyclin I synthase, vascular endothelial growth factor etc. resulted in a reduction of pulmonary arterial pressure in experimental animals with pulmonary hypertension. Major hurdle in current gene therapy for pulmonary hypertension may be the limited duration of transgene expression in lung tissue. However, rather than a limitation, this feature may be a advantage in some situations where expression during only a few weeks may provide both adequate therapeutic efficacy and limited side effects. Development of second-generation gene delivery system will provide new tools to prolong the duration of transgene expression. PMID- 11411129 TI - [Primary pulmonary hypertension]. AB - Primary pulmonary hypertension is a rare and poor prognostic disease. This disease has a pulmonary arteriopathy that is due to unknown cause. Histological study shows medial hypertrophy and fibrosis. Plexogenic arteriopathy is usually observed, but not always. Recently gene mutation was discovered in familial primary pulmonary hypertension. Differential diagnosis is very important because therapies are much different among pulmonary hypertensions due to other causes. In the near future, the mechanism of the disease will be apparent, and therapy must be improved. PMID- 11411130 TI - [Pulmonary hypertension in patients with connective tissue diseases]. AB - Pulmonary hypertension (PH) is said to be frequently associated with connective tissue diseases (CTD). The national surveillance of CTD associated PH (CTD-PH) by the study groups of the Ministry of Health and Welfare (MHW) revealed that mixed connective tissue disease (MCTD) had the highest incidence of PH (5.02%), next to systemic sclerosis (SSc) (2.64%). Patients with CTD-PH could be divided into two groups according to the prognosis: poor prognosis group and fairly good prognosis group. Poor prognosis was associated statistically with polyarthritis and high level of CK. Treatment of CTD-PH was still difficult, but adrenocoticosteroids could be tried in some patients, and drip infusion therapy of prostacyclin seemed very promising, but it has not been accepted by the MHW so far. PMID- 11411131 TI - [Chronic thromboembolic pulmonary hypertension (CTEPH)]. AB - The patients with chronic thromboembolic pulmonary hypertension (CTEPH) usually shows impaired physical activity and poor prognosis, proportional to the degree of pulmonary hypertension. Effort dyspnea and fatigue, the major symptoms, are quite nonspecific, and the physical findings of pulmonary hypertension are easily overlooked until they are far advanced. The most important clue to the diagnosis is perfusion and ventilation lung scan which demonstrates more segmental sized or larger perfusion defects that are normally ventilated. The presence of precapillary pulmonary hypertension should be confirmed by right heart catheterization. Pulmonary angiography is essential not only for diagnosing CTEPH but also for defining thrombus extent and location. The current criteria for considering for surgery include: 1) significant exertional limitation defined as Hugh-Jones > or = III, or NYHA > or = III, 2) pulmonary hypertension defined as PAm > or = 30 mmHg, 3) thrombi located at least as proximally as the lobar level, 4) the absence of significant co-morbid disease, 5) a willingness of the patient and his family to accept the risks of surgery. Although the hemodynamic and symptomatic improvement associated with pulmonary thromboendarterectomy is apparent, the operative mortality is still high, especially in patients with severe hemodynamic disease. The accurate diagnosis and selection criteria for surgery are crucial. PMID- 11411132 TI - [Pulmonary hypertension in patients with congenital and acquired heart diseases]. AB - Pathophysiology of pulmonary hypertension in patients with congenital and acquired heart diseases are described. The broken down of the pulmonary hypertension is left ventricular dysfunction and/or left atrial overloading, and high blood flow in pulmonary circuit with intra-cardiac or intra-vascular shunt. There are pulmonary venous, reactive, and arterial hypertension, related with the mechanism. The worsening of pulmonary venous hypertension induces acute hemodynamic pulmonary edema. The high blood flow in the pulmonary circulation due to large shunt develops Eisenmenger syndrome related with aging. PMID- 11411133 TI - [Chronic hypoxic pulmonary hypertension]. AB - Hypoxic pulmonary vasoconstriction is a self-modulatory mechanism matching local blood flow to local alveolar ventilation. In pathological conditions, it may occur as an acute episode or as a sustained response with vascular remodeling and pulmonary hypertension. Chronic generalized alveolar hypoxia occurs in populations living at high altitude, in human disease states associated with decreased ventilation, or progressive lung destruction. Persistent hypoxic vasoconstriction and the associated vascular structural remodeling are the main mechanisms of the sustained pulmonary hypertension, although other factors, including secondary polycythemia, hypercapnia and increased airways resistance may be involved. Understanding of these mechanisms has increased rapidly and may result in the near future in specific treatment aimed at reversing the structural remodeling and matrix deposition in pulmonary arteries. PMID- 11411134 TI - [Pulmonary hypertension complicating portal hypertension: portopulmonary hypertension]. AB - Portopulmonary hypertension is a condition with a poor prognosis, which is defined as precapillary pulmonary hypertension complicating portal hypertension mainly due to cirrhosis of various etiologies. A mean pulmonary arterial pressure greater than 25 mmHg at rest with a pulmonary capillary wedge pressure less than 15 mmHg and a pulmonary vascular resistance greater than 120 dynes.sec.cm-5, in the setting of the presence of portosystemic shunting has been proposed as hemodynamic criteria for portopulmonary hypertension. Prevalence of pulmonary hypertension ascertained by right cardiac catheterization was 2% among patients with cirrhosis, and reached to 4% particularly among candidates for liver transplantation. Hyperdynamic systemic circulation seen commonly in patients with cirrhosis appeared to be normalized by complication of pulmonary hypertension with a contraction of circulating plasma volume. Long term treatment by epoprostenol administration or nitric oxide inhalation could induce a gradual decline in pulmonary arterial pressure in patients with poor response to acute vasodilator administration. PMID- 11411135 TI - [HIV-associated pulmonary hypertension]. AB - The association between pulmonary hypertension and HIV infection is well established. However, the development and progression of pulmonary hypertension has no relationship with the stage of HIV disease and the history of opportunistic infections. Although the pathogenesis of HIV-associated pulmonary hypertension is not clear, several hypotheses have been proposed. Pulmonary arterial pressures are in the moderate-to-severe range. Plexogenic pulmonary arteriopathy is the most common histopathologic finding. Progressive shortness of breath and pedal edema are the two most common presenting features. Responses to pulmonary vasodilator agents, antiretroviral agents, and anticoagulation therapy are variable. The appearance of unexplained cardiopulmonary symptoms in HIV infected individuals should suggest pulmonary hypertension. Furthermore, HIV testing is recommended in all patients presenting with unexplained pulmonary hypertension, even when they deny HIV risk behaviors. PMID- 11411136 TI - [Drug induced pulmonary hypertension]. AB - Primary pulmonary hypertension (PPH) is a rare, often fatal disease that tends to occur with particular frequency in woman. The factors leading to its development remain enigmatic. A variety of toxins and drugs have been implicated in the pathogenesis of pulmonary hypertension (PH). In the 1960s, the medical community was first alerted to the problem of an epidemic of PH in association with a particular anorexic agent, aminorex. Recent reports have indicated that anorexic drugs may have causative roles in PH. The International Primary Pulmonary Hypertension Study (IPPHS) showed a strong association between PPH and the use of appetite suppressants. But the mechanism by which these anorectic drugs cause PH is unknown. PMID- 11411137 TI - [Persistent pulmonary hypertension of the newborn]. AB - Persistent pulmonary hypertension of the newborn (PPHN) is a cardiopulmonary disorder characterized by systemic arterial hypoxemia secondary to elevated pulmonary vascular resistance with resultant shunting of pulmonary blood flow to the systemic circulation. This disorder can be classified into four forms dependent on the etiology of the pulmonary hypertension: (1) Hypoplastic lung associated with congenital diaphragmatic hernia and oligohydramnions. (2) Primary PPHN (without known causative factor), such as preterm PPHN. (3) Secondary PPHN (with known causative factor), such as meconium aspiration syndrome, birth asphyxia or respiratory distress syndrome. (4) Relative PPHN associated with heart failure of hydrops fetalis or ischemic myocardial dysfunction. Inhalation of nitric oxide, which previously known as endothelial-derived-relaxation-factor, has been studied intensively as therapy for PPHN. PMID- 11411138 TI - [Multiple sclerosis: the recent progress of research and treatment]. AB - In this review, we summarized the recent progress of research and treatment of multiple sclerosis (MS). The immunogenetic study including analyses of HLA polymorphism revealed that there are two distinct subtypes of MS: Asian-type and Western-type. Since MS is a polygenic disease, there must be several susceptible genes other than HLA. A large number of candidate genes are being searched and analyzed for polymorphism. Autoantigens responsible for MS are of great interest. Screening peptide library which bind to MS susceptible genes are expected to provide with new autoantigens. Advance in treatment is also remarkable. Since interferon (IFN)-beta 1b became available, great deal of data concerning IFN beta are accumulated. IFN beta not only reduced clinical exacerbation and MRI brain lesions but also slowed the development of brain atrophy. New treatments include altered peptide ligand therapy and oral tolerization with myelin antigens, which are on the clinical trial. Vaccination with myelin reactive T cells or T cell receptor peptide also attracts a great deal of attention. PMID- 11411139 TI - [Pathogenesis and guidelines for therapy of atopic dermatitis]. AB - Atopic dermatitis (AD) is regarded as one of type 2 helper T cell (Th2)-mediated diseases. However, our data by in situ hybridization elucidated mRNA expressions of both Th1 and Th2 cyotokine in the lesions of AD. Some work revealed a decrease of Th1 mRNA without a change of Th2 mRNA after remission by therapy. Th1 may closely relate to skin lesions of AD. Our work showed a IL-4 and IL-5 production and a elevated expression of high affinity IgE receptor by eosinophils in the lesions of AD. Uptake of allergens may be mediated by the receptor and may lead to a release of cytokines. A guideline for therapy for AD, supported by a Ministry of Health and Welfare in Japan, is published in 1999. In this paper, the guideline was outlined. PMID- 11411140 TI - Standardisation of temperature observed by automatic weather stations. AB - Daily mean, maximum and minimum surface air temperature data were gathered from a network of automatic weather stations (AWS) within the Moor House National Nature Reserve in northern England. Five AWS were installed next to the official Environmental Change Network weather station at Moor House. Data were compared graphically and correction constants were calculated to adjust data from each AWS to the standard of the official station by optimising the concordance correlation coefficient. Each corrected station was re-located next to one of five in-situ stations in and around the reserve, allowing correction of all temperature sensors to a common standard. The mean error associated with measured daily mean, maximum and minimum temperature for each sensor does not exceed +/- 0.2 K. The procedure quantifies a source of systematic measurement error, improving the identification of spatial temperature differences between stations. PMID- 11411141 TI - Ambient air quality at the site of a former manufactured gas plant. AB - Prior to the 1950's, manufactured gas was commercially produced from the pyrolysis of coal, coke, and oil at facilities that are termed manufactured gas plants (MGPs). The constituents of residual coal tar present on many MGP sites are an environmental health concern because of their toxicity and the possibility for their off-site migration via water and air. Atmospheric concentrations of five volatile organic compounds (VOCs, e.g., benzene), sixteen polycyclic aromatic hydrocarbons (PAHs, e.g., naphthalene) and particulate matter less than 10 microns in aerodynamic diameter (PM10) were measured at the site of a former MGP. Air samples were obtained before, during, and after excavation of subterranean coal tar at the site. The results of this investigation indicate that subterranean coal tar was not a primary source of VOCs and PAHs in the local atmosphere before or after remediation of the site. However, excavation, treatment, blending, and transfer of the coal tar during remediation generated concentrations of selected aromatic and semi-volatile organic compounds that were substantially greater than typical ambient levels. In addition, these data suggest that blending and mixing of coal tars could lead to exceedance of the U.S. National Ambient Air Quality Standard for PM10, although additional research is required to fully evaluate this possibility. Nuisance odors associated with the site remediation were likely the result of naphthalene and possibly isomers of xylene. Air pollutant concentrations measured adjacent to the excavation area and at the site perimeter during remediation activities were less than the relevant occupational and environmental exposure limits. PMID- 11411142 TI - Long-term relationship between phosphorus inputs and wetland phosphorus concentrations in a northern Everglades marsh. AB - Assessments of long-term relationships between changes in nutrient inputs and wetland nutrient concentrations can be complicated by fluctuations in other environmental factors as well as by problems typical of long-term monitoring data. Consequently, statistical analysis of these types of data sets requires careful consideration of environmental covariates, potential biases in the monitoring design, and irregularities caused by changes in field sampling protocols. We evaluated the relationship between anthropogenic phosphorus (P) inputs and water-column total P (TP) concentrations in a northern Everglades marsh by statistically analyzing available data collected from several sampling programs over the past 20 years (1978-1997). Canal inputs of agricultural runoff contribute most of the P to the marsh and have produced a zone of enrichment within the marsh during the past few decades. Regression analyses showed that both canal and marsh TP concentrations increased during the 1980s and then decreased in the 1990s. However, the statistical relationship between canal P inputs and marsh TP, while significant, generally was weak except for marsh locations adjacent to the canal. Stronger relationships existed between marsh TP and hydrologic parameters such as marsh water depth, which is controlled by changes in weather patterns and marsh management. In particular, dry conditions during the 1980s may have contributed to observed increases in marsh P concentrations and the movement of a P 'front' further into the marsh. Higher rainfall and water depths and agricultural best management programs initiated during the 1990s have been associated with reduced P concentrations in canal waters entering the marsh. While it is anticipated that this reduction eventually will result in lower marsh TP concentrations, this effect is not yet evident, possibly due to internal loading of P from enriched marsh soils. Our findings illustrate some of the environmental factors that can complicate attempts to develop empirical relationships between P inputs and wetland P concentrations and to use such relationships to forecast changes in marsh concentrations based on past monitoring data alone. PMID- 11411143 TI - Water hardness and sodium trends in Texas aquifers. AB - Median hardness and sodium levels in groundwater were calculated for 244 Texas counties from measurements at 7728 water wells. The data were mapped and analyzed with a geographic information system (GIS). County median hardness levels varied widely, from 4-2304 mg L-1. More than 60% of the counties had hardness medians above 180 mg L-1. County medium sodium concentrations ranged from 6-1170 mg L-1, with more than 90% of those values exceeding 20 mg L-1. There was a significant positive correlation between hardness and sodium concentrations in six of Texas' nine major aquifers. A significant negative correlation between hardness and sodium was observed in two aquifers. Several factors control hardness and sodium variations in Texas aquifers including rock/sediment composition, groundwater chemical evolution, and seepage from nearby formations. Probable human controls include agricultural return flow and pumping-induced saltwater intrusion. PMID- 11411144 TI - Heavy metal accumulation in the gastropod, Cerithium scabridum L., from the Kuwait Coast. AB - An in-depth study on various locations along the Kuwait Coast differentiated by the nature of coastal topography, human habitation and industrialization was made during the year 1995-1997 in relation to the gills of the gastropod, Cerithium scabridum, a bio-indicator of heavy metal pollution. The maximum concentration of heavy metals in the gills of C. scabridum, seawater and particulate matter was observed in Stations III when compared to the metal concentrations in the other seven stations. Co-efficient Variance (CV) revealed an increase in the accumulation of Cu and Pb in gills, seawater and particulate matter in winter than in summer. However, the CV was noted to be lower in winter than in summer in the case of Cr and V, in gills of C. scabridum and Ni in the case of seawater samples. Further, by ANOVA, except V, significant relationships were noted between the heavy metals (Cu, Cr, Pb and Ni) and various locations in the gills of C. scabridum. PMID- 11411145 TI - Exposure assessment to mercury vapor in chloralkali industry. AB - The aim of the study was to develop a sampling method aimed at individual sampling of mercury vapor and subsequent individual exposure assessment of the worker. Hopcalite available from Inowroclawskie Zaklady Chemiczne (in Poland) was found useful for mercury vapor sampling in active and passive individual samplers. However the sampling rate determined for passive sampler in steady state laboratory chamber cannot be used under fluctuating conditions of mercury vapor concentration and air movement typical for field measurement. In order to check comparativeness of passive and active sampling methods, samples were taken in the same time and individual samplers fixed to worker's clothes were used. Mercury vapor concentration was measured at two chloralkali industries in Poland and the results were presented. Excessive exposure in both industries was confirmed by finding elevated mercury concentration in urine samples from the workers. PMID- 11411147 TI - A random walk through models of nonlinear clustering. AB - A few simple models of the mass function of collapsed objects are presented. The emphasis is on apparently unrelated models which end up giving the same answer for the number density and merger histories of virialized clumps. Models of the spatial distribution of the clumps and how they can be used to model the spatial distribution of the mass are briefly discussed. PMID- 11411146 TI - A stream visual assessment protocol (SVAP) for riparian landowners. AB - A user-friendly Stream Visual Assessment Protocol (SVAP) was recently developed in a joint effort by the Natural Resources Conservation Service (NRCS) of the U.S. Department of Agriculture and the University of Georgia. SVAP was designed to be an introductory screening-level assessment method for people unfamiliar with stream assessments. It was designed for use by NRCS field staff who work with agricultural landowners. NRCS is in a key position to influence conservation practices since the organization works with private stakeholders, maintaining more than 2000 field offices throughout the U.S. with a central office in each state. The SVAP measures a maximum of 15 elements and is based on visual inspection of the physical and biological characteristics of instream and riparian environments. Each element is assigned a numerical score relative to reference conditions and an overall score for the stream reach is calculated. A qualitative description of the stream reach is made based on overall numerical score. While SVAP is not intended to replace more robust stream assessment protocols, it provides quick and reliable information for use in NRCS farm assistance programs. It is also an educational tool through which landowners can learn about conservation of aquatic resources. An abridged copy of SVAP is attached as an appendix to this article and the complete document can be found on the web at http://www.ncg.nrcs.usda.gov/tech_notes.html. PMID- 11411148 TI - Dark matter caustics. AB - The late infall of cold dark matter onto an isolated galaxy such as our own produces flows with definite velocity vectors at any physical point in the galactic halo. It also produces caustics which are places where the dark matter density is very large. The outer caustics are topological spheres whereas the inner caustics are rings. The self-similar model of galactic halo formation predicts that the caustic ring radii a(n) follow the approximate law a(n) approximately 1/n. In a recent study of 32 extended and well-measured galactic rotation curves, we found evidence for this law. PMID- 11411149 TI - Nonlinear gravitational growth inside and outside the standard cosmology. AB - We reconsider the problem of nonlinear structure formation inside and outside General Relativity (GR), both in the weakly and strongly nonlinear regime. We show how these regimes can be explored observationally through clustering of high order cumulants and through the epoch of formation, abundance and clustering of collapse structures, using Press and Schechter (1974, Astrophys. J. 187: 425-438) formalism and its extensions. PMID- 11411150 TI - An attempt to do without dark matter. AB - The discrepancy between dynamical mass measures of objects such as galaxies and the observed distribution of luminous matter in the universe is typically explained by invoking an unseen "dark matter" component. Dark matter must necessarily be nonbaryonic. We introduce a simple hypothesis to do away with the necessity for dark matter by introducing an additional non-gravitational force coupled to baryon number as a charge. We compare this hypothesis to Milgrom's modified Newtonian dynamics. The model ultimately fails when confronted with observation, but it fails in an interesting way. PMID- 11411151 TI - A new angle on gravitational clustering. AB - A new approach to gravitational instability in large-scale structure is described, where the equations of motion are written and solved as in field theory in terms of Feynman diagrams. The basic objects of interest are the propagator (which propagates solutions forward in time), the vertex (which describes nonlinear interactions between waves) and a source with prescribed statistics which describes the effect of initial conditions. We show that loop corrections renormalize these quantities, and discuss applications of this formalism to a better understanding of gravitational instability and to improving nonlinear perturbation theory in the transition to the nonlinear regime. We also consider the role of vorticity creation due to shell-crossing and show using N body simulations for which at small (virialized) scales the velocity field reaches equipartition, that is, the vorticity power spectrum is about twice the divergence power spectrum. PMID- 11411152 TI - Emergence of anomalous distributions in disordered systems. AB - The present day non-gaussian distribution of mass density of the universe evolved from an initial gaussian distribution in the presence of nonlinear interactions. We discuss an analog in disordered condensed matter system where increasing the disorder changes the distribution of conductances from a gaussian at weak disorder to a log-normal at strong disorder. The highly asymmetric "one-sided" log-normal distribution in the intermediate crossover regime can be understood as a simple hybrid of these two limiting distributions. PMID- 11411153 TI - The onset of nonlinearity in cosmological structure. AB - We discuss the progression of growth of cosmological structure, from the quasilinear evolution of nearly Gaussian fluctuations on large scales into highly non-Gaussian, strongly nonlinear structure on small scales. A systematic development in perturbation theory describes the first departures from homogeneity but fails to reproduce the fully nonlinear results. Physical insight, conceptual models, and symmetries are useful in the strong clustering regime. A phenomenological model with input information from the quasilinear regime provides enticing results for the strongly nonlinear regime. PMID- 11411154 TI - The transition to nonlinearity and new constraints on biasing. AB - We present two new dynamical tests of the biasing hypothesis. The first is based on the amplitude and the shape of the galaxy-galaxy correlation function, xi g(r), where r is the separation of the galaxy pair. The second test uses the mean relative peculiar velocity for galaxy pairs, v12(r). This quantity is a measure of the rate of growth of clustering and it is related to the two-point correlation function for the matter density fluctuations, xi (r). Under the assumption that galaxies trace the mass (xi g = xi), the expected relative velocity can be calculated directly from the observed galaxy clustering. The above assumption can be tested by confronting the expected v12 with direct measurements from velocity-distance surveys. Both our methods are checked against N-body experiments and then compared with the xi g(r) and v12 estimated from the APM galaxy survey and the Mark III catalogue, respectively. Our results suggest that cosmological density parameter is low, omega m approximately 0.3, and that the APM galaxies trace the mass at separations r > or = 5 h-1, where h is the Hubble constant in units of 100 km s-1 Mpc. The present results agree with earlier studies, based on comparing higher-order correlations in the APM with weakly nonlinear perturbation theory. Both approaches constrain the linear bias factor to be within 20% of unity. If the existence of the feature we identified in the APM xi g(r)--the inflection point near xi g = 1--is confirmed by more accurate surveys, we may have discovered gravity's smoking gun: the long awaited "shoulder" in xi, generated by gravitational dynamics and predicted by Gott and Rees 25 years ago. PMID- 11411155 TI - Peculiar velocity surveys: optimal moments analysis. AB - A new formalism to analyze peculiar velocity surveys is presented. Results from these surveys are shown to be dominated by small-scale noise, aliasing, and incomplete cancellations. The formalism allows us to filter out the signal from scales that are not of interest and thus provides us with a clean signal that probe large scales. We use maximum likelihood techniques to analyze the filtered data and compare it to the analysis of the full dataset. The filtered analysis gives a much better parameter estimation than the full analysis, leading us to conclude that, indeed, the large-scale signal is obscured by small-scale noise. PMID- 11411156 TI - Non-gaussianity versus nonlinearity of cosmological perturbations. AB - Following the discovery of the cosmic microwave background, the hot big-bang model has become the standard cosmological model. In this theory, small primordial fluctuations are subsequently amplified by gravity to form the large scale structure seen today. Different theories for unified models of particle physics, lead to different predictions for the statistical properties of the primordial fluctuations, that can be divided in two classes: gaussian and non gaussian. Convincing evidence against or for gaussian initial conditions would rule out many scenarios and point us toward a physical theory for the origin of structures. The statistical distribution of cosmological perturbations, as we observe them, can deviate from the gaussian distribution in several different ways. Even if perturbations start off gaussian, nonlinear gravitational evolution can introduce non-gaussian features. Additionally, our knowledge of the Universe comes principally from the study of luminous material such as galaxies, but galaxies might not be faithful tracers of the underlying mass distribution. The relationship between fluctuations in the mass and in the galaxies distribution (bias), is often assumed to be local, but could well be nonlinear. Moreover, galaxy catalogues use the redshift as third spatial coordinate: the resulting redshift-space map of the galaxy distribution is nonlinearly distorted by peculiar velocities. Nonlinear gravitational evolution, biasing, and redshift space distortion introduce non-gaussianity, even in an initially gaussian fluctuation field. I investigate the statistical tools that allow us, in principle, to disentangle the above different effects, and the observational datasets we require to do so in practice. PMID- 11411157 TI - The cosmological mass function in the Zel'dovich approximation. AB - The Press-Schechter theory of the cosmological mass function and its modifications allow constraint of cosmological scenarios of structure formation. Recently, a few new models that explored the influence of anisotropic collapse on the shape of the mass function have been suggested. I discuss in more detail a particular model that assumes a fluid particle becomes part of a gravitationally bound halo when the smallest eigenvalue of the deformation tensor of the filtered initial density field reaches a certain threshold (like the filtered density contrast reaches the threshold in the Press-Schechter formalism). Choosing the smallest eigenvalue guarantees that the fluid particle in question experiences collapse along all three axes. The model shows a better agreement with the N-body simulations than with the standard Press-Schechter model. PMID- 11411158 TI - Lensing of the CMB: non-Gaussian aspects. AB - We compute the small angle limit of the three- and four-point function of the cosmic microwave background (CMB) temperature induced by the gravitational lensing effect by the large-scale structure of the universe. We relate the non Gaussian aspects presented in this paper with those in our previous studies of the lensing effects. We interpret the statistics proposed in previous work in terms of different configurations of the four-point function and show how they relate to the statistic that maximizes the S/N. PMID- 11411159 TI - Cosmic statistics of statistics: N-point correlations. AB - The fully general calculation of the cosmic error on N-point correlation functions and related quantities is presented. More precisely, the variance caused by the finite volume, discreteness, and edge effects is determined for any estimator which is based on a general function of N-tuples, such as multi-point correlation functions and multi-spectra. The results are printed explicitly for the two-point correlation function (or power-spectrum), and for the three-point correlation (or bispectrum). These are the most popular statistics in the study of large-scale structure, yet the general calculation of their variance has not been performed until now. PMID- 11411160 TI - Introduction. Unity of knowledge, circa 2000. PMID- 11411161 TI - The anterior cingulate cortex. The evolution of an interface between emotion and cognition. AB - We propose that the anterior cingulate cortex is a specialization of neocortex rather than a more primitive stage of cortical evolution. Functions central to intelligent behavior, that is, emotional self-control, focused problem solving, error recognition, and adaptive response to changing conditions, are juxtaposed with the emotions in this structure. Evidence of an important role for the anterior cingulate cortex in these functions has accumulated through single neuron recording, electrical stimulation, EEG, PET, fMRI, and lesion studies. The anterior cingulate cortex contains a class of spindle-shaped neurons that are found only in humans and the great apes, and thus are a recent evolutionary specialization probably related to these functions. The spindle cells appear to be widely connected with diverse parts of the brain and may have a role in the coordination that would be essential in developing the capacity to focus on difficult problems. Furthermore, they emerge postnatally and their survival may be enhanced or reduced by environmental conditions of enrichment or stress, thus potentially influencing adult competence or dysfunction in emotional self-control and problem-solving capacity. PMID- 11411162 TI - How to unify knowledge. Keynote address. AB - The central question of general scholarship is whether all of knowledge is intrinsically consilient, that is, whether it can be united by a continuous skein of cause-and-effect explanation and across levels of increasingly complex organization. The answer lies in the recognition that the traditional line separating the great branches of learning (natural sciences, social sciences, and humanities) is not a barrier or even a line, but rather a broad domain of poorly explored material phenomena now being cooperatively entered from all sides. The key to bridge-building is the discovery and analysis of human nature, which consists of the epigenetic rules--the hereditary regularities in mental development. Their study is now well under way, at the biological, social science, and humanities levels. Examples of epigenetic rules include the origin of color vocabularies, incest avoidance, optimum arousal in artistic design, and response to the natural environment. The convergence of the great branches of learning is mostly an empirical process, light on formal logic and theory in its initial stages. The value of the consilience program is that at long last we appear to have acquired the means either to establish the intrinsic truth of the fundamental unity of knowledge, or to discard it. I will argue the likelihood of its existence and early establishment. PMID- 11411163 TI - Language/culture/mind/brain. Progress at the margins between disciplines. AB - At the forefront of research on language are new data demonstrating infants' strategies in the early acquisition of language. The data show that infants perceptually "map" critical aspects of ambient language in the first year of life before they can speak. Statistical and abstract properties of speech are picked up through exposure to ambient language. Moreover, linguistic experience alters infants' perception of speech, warping perception in a way that enhances native language speech processing. Infants' strategies are unexpected and unpredicted by historical views. At the same time, research in three additional disciplines is contributing to our understanding of language and its acquisition by children. Cultural anthropologists are demonstrating the universality of adult speech behavior when addressing infants and children across cultures, and this is creating a new view of the role adult speakers play in bringing about language in the child. Neuroscientists, using the techniques of modern brain imaging, are revealing the temporal and structural aspects of language processing by the brain and suggesting new views of the critical period for language. Computer scientists, modeling the computational aspects of childrens' language acquisition, are meeting success using biologically inspired neural networks. Although a consilient view cannot yet be offered, the cross-disciplinary interaction now seen among scientists pursuing one of humans' greatest achievements, language, is quite promising. PMID- 11411164 TI - Biological constraint, cultural variety, and psychological structures. AB - Although biological processes bias humans to develop particular cognitive, affective, and behavioral forms, the cultural context of growth shapes these forms in particular ways. Psychologists have been indifferent to the nature of the mental structures that mediate the varied psychological functions that are the usual target of inquiry. This paper argues that schemata for perceptual events, motor programs, and semantic networks are distinct, although interdependent, forms that rest on different neurophysiologies. The biological constraints are weakest on the semantic networks that are influenced by the history, economy, religion, geography, and social structure of the society. These factors influence how cultures classify names for emotions, categories of self membership, and popular metaphors for human nature. One class of schemata is derived from changes in body tone. Temperamental variation in the susceptibility to changes in body tone has relevance for understanding personality and a vulnerability to anxiety disorders. PMID- 11411165 TI - Prolegomenon to a general biology. AB - This chapter is a slightly modified version of Chapter 1 of my book, Investigations. I discuss "autonomous agents," the origin of life, autocatalytic sets of polymers, work cycles, puzzles-about evolutionary theory, and the birth of a general biology dealing with biospheres anywhere in the universe. PMID- 11411166 TI - Toward a biology of personality and emotion. AB - For most of this past century, scholarship on the topics of personality and emotion has emerged from the humanities and social sciences. In the past decade, a remarkable change has occurred in the influence of neuroscience on the conceptualization and study of these phenomena. This article argues that the categories that have emerged from psychiatric nosology and descriptive personality theory may be inadequate, and that new categories and dimensions derived from neuroscience research may produce a more tractable parsing of this complex domain. The article concludes by noting that the discovery of these biological differences among individuals does not imply that the origins of these differences lie in heritable influences. Experiential shaping of the brain circuitry underlying emotion is powerful. The neural architecture provides the final common pathway through which culture, social factors, and genetics all operate together. PMID- 11411167 TI - Exploring the biology of socialization. AB - How will the social sciences take advantage of the revolution that has taken place in biology during the past two decades? Over the last fifteen years, neuroimaging has allowed the study of human cognition and emotion within psychology to achieve close alliances with biology through the development of cognitive and affective neuroscience. There is little doubt that a similar alliance between psychology and biology will occur in the domain of human brain development. In principle, understanding how the human brain is organized by experience (epigenetic rules) and how societies instruct their young could produce a link between natural and social science. The late David C. McClelland sought methods to base the social sciences on psychological ideas. McClelland sought to connect the values of achievement and power as coded from children's readers and popular ballads to societal economic growth and conflict. These efforts lacked knowledge of brain mechanisms of memory and attention and an understanding of the role of experience in organizing brain circuitry. Understanding of cognitive and brain systems related to knowledge and action may allow a new approach to forging connections between individual minds and social behavior. PMID- 11411168 TI - A polytheistic conception of the sciences and the virtues of deep variety. PMID- 11411169 TI - Science, culture, meaning, values: a dialogue. PMID- 11411170 TI - Science in the liberal arts curriculum. PMID- 11411171 TI - K-12 science education as the road to consilient curricula. AB - We begin with the absurdity of ninth-grade biology to bewail the fate of high school science education as one component of a wholly inadequate K-12 education. Our proposal for a coherent physics-chemistry-biology core curriculum for all high school students leads naturally to seeking the connections between the sciences, the sciences and mathematics, and indeed to the social sciences and the humanities. Our goal is literacy--scientific and humanistic--as the sensible goal for a 21st century high school graduate. PMID- 11411172 TI - Integrated learning. Passing fad or foundation for the future? AB - On the surface, higher education has served the nation well. An increasing percentage of adults have attended college. Information is widely and freely available to most citizens; the economy is steady; and technological developments appear to offer ever-increasing improvements in the quality of life. Looking just beneath the surface, though, one realizes that knowledge is fragmented, access to knowledge is not universal, and there are no guarantees that past successes will sustain the future. Thus, it is worth examining how higher learning has contributed to the general prosperity and how education might evolve to address society's evident problems. This essay discusses the core approach of liberal arts in the context of the value--and obstacles--of integration across the traditional disciplines. Critical to the natural sciences is a firm grounding in the central importance of rigorous evidence, while the humanities keep us rooted in the importance of human values. Seeking linkages and connections between these realms is the lively challenge of this conference. The details of curricular design will be of less interest at this juncture than locating the critical issues and discussing how education might serve the goal of unifying knowledge and learning. PMID- 11411173 TI - Some practical suggestions for teaching science in the liberal arts. AB - At the undergraduate level, science is held at a distance from the liberal arts by common agreement among professors from both sides of a wall erected for them by the Enlightenment some centuries ago. There is today no intellectual argument put forth for maintaining the wall; the arguments in its defense are largely cultural and political and therefore in the final analysis fiscal, if not venal. Here are some concrete suggestions for how that wall might be brought down. PMID- 11411174 TI - From molecules to mind. Stress, individual differences, and the social environment. AB - The social and physical environments in which we live have an enormous impact on our physiology and behavior and influence the process of adaptation, or "allostasis." Genes, early development, adult experiences, life style, and stressful life experiences all contribute to the way the body adapts to a changing environment; and these factors all help to determine the cost to the body, or "allostatic load." Studies of these processes involve the disciplines of biology and psychology, but they are incomplete without the input from other fields, such as cultural anthropology, economics, epidemiology, political science, and sociology. These fields provide a description and analysis of the social and cultural institutions and economic forces that affect individual human health. Specific examples of shared concepts and terminology are given to illustrate progress towards consilience in the study of socioeconomic determinants of health. PMID- 11411175 TI - Nature, nurture, and the disunity of knowledge. AB - The Human Genome Project and the tools of modern molecular biology bring enormous promise for the understanding of human biology. Juxtaposed, however, is a conceptual stagnation reflected in the continued nature/nurture debate. More sophisticated models reflecting the inevitable interdependence of gene and environment are essential if we are to realize the potential offered by today's technological advances. PMID- 11411176 TI - Do people make environments or do environments make people? AB - This article discusses the influence of people's genetic make-up on their mental states of happiness and depression. Contrary to the conventional wisdom, great fortune does not guarantee happiness; neither does great misfortune assure depression. Emotional states are surprisingly immune to "objective" social circumstances. A biological basis for this relative immunity is that people possess biological set points for these emotional states, rendering the effects of most life events transitory. Genotypes also have indirect effects. People react differently to psychological stressors depending on their genotypes. A susceptible person may succumb to depression, whereas a resilient person may remain unaffected. People also expose themselves to different social environments. Exposure to controllable life events is partly a result of genetic predispositions. Consilience requires that this biological individuality be considered in any understanding of human behavior, including the pursuit of happiness. PMID- 11411177 TI - The smoke detector principle. Natural selection and the regulation of defensive responses. AB - Defenses, such as flight, cough, stress, and anxiety, should theoretically be expressed to a degree that is near the optimum needed to protect against a given threat. Many defenses seem, however, to be expressed too readily or too intensely. Furthermore, there are remarkably few untoward effects from using drugs to dampen defensive responses. A signal detection analysis of defense regulation can help to resolve this apparent paradox. When the cost of expressing an all-or-none defense is low compared to the potential harm it protects against, the optimal system will express many false alarms. Defenses with graded responses are expressed to the optimal degree when the marginal cost equals the marginal benefit, a point that may vary considerably from the intuitive optimum. Models based on these principles show that the overresponsiveness of many defenses is only apparent, but they also suggest that, in specific instances, defenses can often be dampened without compromising fitness. The smoke detector principle is an essential foundation for making decisions about when drugs can be used safely to relieve suffering and block defenses. PMID- 11411178 TI - Immunoalienation. A behavioral analysis of the immune system. AB - Immunoalienation denotes the process through which the definition of self, as defined by the immune system, progressively deviates from the genetically determined definition of self. The mechanisms responsible for immunoalienation are discussed in light of the available evidence. The different kinds of immunoalienation are analyzed, and their theoretical as well as practical implications to disease, tumors, and transplant biology are briefly discussed. PMID- 11411179 TI - Emotion, cognition, and the human brain. PMID- 11411180 TI - The depressed suicidal patient. Assessment and treatment. AB - Depressive disorders are associated with significant psychosocial impairment and disability. Depression should be thoroughly evaluated, as should current and past suicidality and potential risk factors for suicide. Mortality by suicide characterizes the course of major affective disorders in approximately 15% of those suffering from these illnesses. Several neurobiological correlates of suicidality have been discovered. Treatment of depression with suicidality may involve hospitalization, pharmacotherapy, electroconvulsive therapy, and psychotherapy. Special populations include children and adolescents, the elderly, medically ill patients, patients with comorbid personality disorders, and patients with comorbid substance abuse disorders. Clinicians encountering patients with depressive disorders should be proficient in the assessment and treatment of depression with suicidality. PMID- 11411181 TI - Assessment and treatment of the youthful suicidal patient. AB - In this review, a framework for the assessment of suicidal risk in the adolescent is described, based on existing epidemiological and clinical studies. The assessment of risk can then be used to determine the immediate disposition, intensity of treatment, and level of care. Furthermore, the assessment of psychiatric and psychological characteristics of the individual and family, as well as the motivation and precipitants for the suicidal episode, can be used to target areas of vulnerability and thereby help the patient reduce the risk of recurrent suicidal behavior. The approach to treatment, guided by the assessment, uses a model of suicidal behavior that is based on our clinical experience and the few extant clinical trials of the treatment of suicidal behavior. Recommended interventions involve treatment of psychopathology; amelioration of cognitive distortion and difficulties with social skills, problem-solving, and affect regulation; and family psychoeducation and intervention. Given the chronic and recurrent nature of the conditions associated with adolescent suicide attempts, a long-term care plan involving both continuation and maintenance treatment is advocated. Further research is necessary to identify the most effective approaches to the treatment of adolescent suicide attempters. PMID- 11411182 TI - Suicide in elders. AB - Older persons in the United States are at higher risk for suicide than any other segment of the population. The epidemiology and risk factors for suicide in later life and the most promising approaches to its prevention are reviewed. Available data suggest that psychiatric and physical illnesses, functional impairment, personality traits of neuroticism and low openness to experience, and social isolation are important correlates of late-life suicide. Affective illness is the risk factor with the strongest association. As treatable conditions in most cases, mood disorders are critical targets for preventive interventions. Because 70% of older adults who committed suicide saw their primary care provider within 30 days of death, the primary care setting is an important venue for intervention. Mood disorders are common in primary care practice, but often go undiagnosed and inadequately treated. One important approach to late-life suicide prevention, therefore, is to optimize the ability of primary care providers to diagnose and treat late-life mood disorders and suicidality effectively. Other elders at high risk have no active relationship to primary care. Strategies designed to identify this group and provide them with preventive services through outreach to the community have shown promise as late-life suicide prevention measures as well. PMID- 11411183 TI - Suicide: what is in the clinician's mind? AB - The assessment and treatment of the suicidal patient challenge all clinicians, regardless of training or theoretical orientation, to draw upon their knowledge and judgment to formulate the best treatment plan possible. As great as the influence of demographics, genetics, and psychobiology are, what is in the mind of the clinician, whether it is anxiety, confidence, knowledge, or uncertainty, will determine the fate of the suicidal patient. PMID- 11411184 TI - Treating the suicidal patient. Basic principles. AB - Successful psychotherapy with suicidal patients requires an emotionally full, active engagement from the therapist with his patient. Emphasis is on the real relationship, not the transference, and the therapist must be available to the patient as a sturdy, reliable object with whom to identify. The therapist's attitude must be loving, not neutral; the alliance is built upon the therapist's devotion to the patient's growth and the development of the attributes necessary for successful autonomous adult functioning. Patients require emotional containment and support, assistance in modulating painful affect, validation, education, help with reality testing, and kindly limit setting. Countertransference reactions must be expected and kept in check, so that the therapist does not get in the patient's way as he tries to build up the faulty mental structures that got him into trouble in the first place. PMID- 11411185 TI - Recognizing and responding to a suicide crisis. AB - Data from therapists who were treating 26 patients when they committed suicide were utilized to identify signs that warned of a suicide crisis. Three factors were identified as markers of the suicide crisis: a precipitating event; one or more intense affective stats other than depression; and at least one of three behavioral patterns: speech or actions suggesting suicide, deterioration in social or occupational functioning, and increased substance abuse. Problems in communication between patient and therapist, often originating in therapeutic anxiety over the patient's possible suicide, were identified as factors interfering with crisis recognition. Evaluation of the identified affects and behaviors may help therapists recognize a suicide crisis. PMID- 11411186 TI - When to hospitalize patients at risk for suicide. AB - The decision to hospitalize patients at imminent risk for suicide requires careful assessment of risk factors, including sociodemographic, psychiatric, general medical, and mental status considerations. Assessment of these risk factors is essential and can usually be accomplished in a straightforward manner, although clinical obstacles sometimes make assessment more difficult. Key issues regarding imminent suicide risk are intent and means, severity of psychiatric illness, the presence of psychosis or hopelessness, lack of personal resources, and older age among men. Once the decision to hospitalize is made, reasonable care should be taken to assure that the patient is transported to the treating hospital safely and is not left alone. On arrival at the hospital, the patient should be closely monitored, and reasonable precautions must be taken to assure the patient's safety at all times, especially during the first few days. Proper assessment, monitoring, and treatment of patients at imminent risk for suicide save lives. A person who is determined to kill himself/herself will probably prevail despite our best efforts. However, most people who are imminently suicidal at one time will feel very differently at some later time. PMID- 11411187 TI - Suicide and the media. AB - Evidence continues to amass on the significant impact of media coverage on suicide. The research literature on the impact of news reports of nonfictional suicides as well as fictional suicide stories is reviewed in order to determine the nature and scope of the influence of the mass media on suicide. The current review, building upon earlier reviews, is limited to English language publications or English translations of articles and/or abstracts. The interactive factors that may moderate the impact of media stories are also reviewed. Such interactive factors include characteristics of the stories (agent), individuals' attributes (host), and social context of the stories (environment). Recommendations are presented for the reporting of suicide stories, which may minimize the risk of imitative suicides. The media's positive role in educating the public about risks for suicide and shaping attitudes about suicide is emphasized. In summary, the existence of suicide contagion no longer needs to be questioned. We should refocus our research efforts on identifying which particular story components promote contagion under which circumstances and which components are useful for preventive programming. PMID- 11411188 TI - Firearms and suicide. AB - The evidence linking firearms in the home to risk for suicide is reviewed. These data come from epidemiological, case-control, quasiexperimental, and prospective studies. The convergent finding from this wide range of studies is that there is a strong relationship between firearms in the home and risk for suicide, most firmly established in the United States. PMID- 11411189 TI - Treating the suicidal patient with bipolar disorder. Reducing suicide risk with lithium. AB - Bipolar disorder is associated with increased mortality because of complications of commonly comorbid substance use and stress-sensitive medical disorders as well as accidents and very high rates of suicide. Long-term lithium treatment may be associated with reduced suicidal risk. We review and summarize findings that help to quantify relationships between the presence versus the absence of lithium maintenance and suicides or attempts in patients with bipolar or other major affective disorders. Results from 33 studies (1970-2000) yielded 13-fold lower rates of suicide and reported attempts during long-term lithium treatment than without it or after it was discontinued. Although greatly reduced, these rates remain above those estimated for the general population. Evidence for substantial, if incomplete, protection against suicide with lithium is supported by more compelling evidence than that for any other treatment provided for patients with mood disorders. Studies of commonly used, but incompletely evaluated, alternative treatments are required, and further protection against premature mortality can be anticipated with better protection against bipolar depression. PMID- 11411190 TI - Treatment of suicidality in schizophrenia. AB - Between 4 and 13% of people with schizophrenia commit suicide and between 25 and 50% make a suicide attempt, a reflection of the devastating toll this syndrome takes on the quality of life, that is, the subjective and objective sense of well being. Many risk factors for suicide in schizophrenia have been identified, the most important of which are previous suicide attempts, depression, hopelessness, substance abuse, and male gender. Insight into having a serious mental illness and less severe cognitive impairment are also associated with increased risk for suicide in schizophrenia, most likely when accompanied by feelings of hopelessness. Typical neuroleptic drugs have not been shown to reduce the risk of suicide. However, several types of evidence suggest that clozapine, an atypical antipsychotic drug, appreciably reduces the suicide attempt and completion rates in schizophrenia and schizoaffective disorder, perhaps by as much as 75-85%. Other atypical antipsychotic drugs may have a similar effect, but direct evidence is lacking. Improvement in positive and negative symptoms, reduced extrapyramidal side effects (EPS), a direct antidepressant action, improved cognitive function, and improved compliance may contribute to reduced suicidality. The International Suicide Prevention Trial (InterSePT) is a large prospective, randomized study intended to compare the effectiveness of clozapine with that of olanzapine in reducing suicide and suicide-related events in schizophrenic and schizoaffective patients. Some information about suicidality in the patient sample is reported here. PMID- 11411191 TI - Borderline personality disorder. Suicidality and self-mutilation. AB - Epidemiological and neurobiological perspectives about suicidality in borderline patients are described, highlighting how self-destructive and seemingly suicidal acts are their "behavioral specialty." Principles for management, including the need for a primary clinician to oversee safety, and the pros and cons of both "contracting for safety" and hospitalization are presented. Clinical material is used to illustrate an approach that involves minimal initiative to rescue by therapists, a readiness to comply with patients' wishes for protection (the principle of false submission), and very active interpretation. This approach is contrasted with those advocated by Linehan and Kernberg. PMID- 11411192 TI - Treating the substance-abusing suicidal patient. AB - Studies concerning the treatment of substance-abusing suicidal patients are scarce despite the frequent presence of suicidal behavior among this population. Indeed, suicidality (ideation or behavior) is generally an exclusion criterion for participation in treatment studies of subjects with alcohol or drug abuse. Consequently, to date, little is known about the optimal treatment of this population. The first study involving substance-abusing suicidal patients was an open-label trial conducted in the early 1990s. This study involved 12 patients, all of whom demonstrated recent suicidal ideations and had made a lifetime suicide attempt. The results of that open-label study demonstrated significant within-group improvement in both depressive symptoms (including suicidal ideations) and level of drinking. However, substantial residual depressive symptoms and drinking persisted at the end of the trial. Also, because no placebo control group was utilized, the authors of that study could not rule out the possibility that the apparent therapeutic effect from fluoxetine was the result of the placebo effect. To date, only one double-blind, placebo-controlled study of subjects with alcohol or substance abuse has included substantial numbers of suicidal patients. The study involved 51 subjects, of whom 20 (39%) had made a suicide attempt in the current depressive episode, 31 (61%) had made a suicide attempt in their lifetime, and 46 (90%) had reported suicidal ideations in the week before hospitalization. The results of that double-blind, placebo-controlled study suggest that fluoxetine was effective in decreasing but not eliminating both the depressive symptoms (including suicidal ideations) and the level of alcohol consumption among a study group of subjects with comorbid major depressive disorder and alcohol dependence, many of whom displayed suicidal ideations. A secondary data analysis from that study suggested that cigarette smoking is also significantly decreased by fluoxetine, but the magnitude of the decrease is limited and few of these patients totally quit smoking with fluoxetine treatment alone. Another secondary data analysis from that study suggested that marijuana smoking was also significantly decreased in a subgroup of subjects who demonstrated cannabis abuse and that the magnitude of this improvement was robust. A third secondary data analysis from that study suggested that cocaine abuse acts as a predictor of poor outcome for both depressive symptoms (including suicidality) and level of alcohol use in this population. The results of a 1-year naturalistic follow-up study involving the patients from that study suggest that the benefits of fluoxetine in decreasing depressive symptoms and level of drinking persist 1 year after entering the treatment program. To date, no other double-blind, placebo-controlled studies involving substantial numbers of substance-abusing suicidal patients have been reported to either confirm or refute these findings. Further studies are clearly warranted to evaluate the efficacy of various pharmacotherapeutic agents and various psychotherapies in the treatment of substance-abusing suicidal patients. PMID- 11411193 TI - Treating impulsivity and anxiety in the suicidal patient. AB - It has been found that while over 90% of people who commit suicide have a psychiatric illness at the time, over 50% are under active psychiatric or mental health care. How can suicide risk be detected and preventive treatment provided? Both communication of suicidal ideation or intent and prior suicide attempts have been shown to be risk factors, which should be assessed in patients with psychiatric illness. Research shows that suicidal ideation is often not communicated to professionals or is denied by patients just prior to suicide and, when present, is often useful not as an immediate risk factor, but as a chronic risk factor. Suicide attempts predict a 10-30% occurrence of suicide over 10 years, but often do not indicate immediate risk. Recent research has shown that impulsiveness and severe anxiety, panic attacks, and agitation comorbid with depression are often immediate suicide risk factors that are potentially modifiable if recognized and treated urgently with effective medications and watchful support. PMID- 11411194 TI - Ubiquitin-mediated degradation of cellular proteins: why destruction is essential for construction, and how it got from the test tube to the patient's bed. AB - Between the 1960s and 1980s, the main focus of biological research was nucleic acids and the translation of the coded information into proteins. Protein degradation was a neglected area and regarded by many as a scavenger, non specific and end process. While it was known that proteins are turning over, the large extent and high specificity of the process--where distinct proteins have half-lives that range from a few minutes to several days--have not been appreciated. The discovery of the lysosome by Dr. Christian de Duve did not change this view significantly, as this organelle is involved mostly in the degradation of extra- and not intracellular proteins, and it was clear that lysosomal proteases, similar to those of the gastrointestinal tract, cannot be substrate specific. The discovery of the complex cascade of the ubiquitin pathway has changed this view dramatically. It is now clear that degradation of cellular proteins is a highly complex, temporally controlled, and tightly regulated process that plays major roles in a broad array of basic pathways during cell life and death. With the multitude of substrates targeted and processes involved, it is not surprising that aberrations in the pathway have been recently implicated in the pathogenesis of many diseases, certain malignancies and neurodegeneration among them. Degradation of a protein via the ubiquitin pathway involves two successive steps: a) conjugation of multiple ubiquitin moieties to the substrate, and b) degradation of the tagged protein by the downstream 26S proteasome complex with release of free and re-utilizable ubiquitin. Despite intensive research, the unknown still exceeds what we currently know on intracellular protein degradation and major key problems remain unsolved. Among these are the modes of specific and timed recognition of the myriad substrates of the system and the nature of the mechanisms that underlie aberrations in the system and pathogenesis of diseases. PMID- 11411195 TI - Safety and immunogenicity of a new mammalian cell-derived recombinant hepatitis B vaccine containing Pre-S1 and Pre-S2 antigens in adults. AB - BACKGROUND: Previous data showed that new recombinant hepatitis B virus vaccine, which contains the S-protein component of the HBV surface together with the Pre S1 and Pre-S2, is considerably more immunogenic than a second-generation recombinant HBV vaccine. OBJECTIVES: To compare the immunogenicity and safety of a novel recombinant HBV vaccine S1, Pre-S1 and Pre-S2 protein components of the hepatitis B surface antigen--BioHep, 10 micrograms dose, to a licensed vaccine containing only the S-protein component--Engerix-B, 20 micrograms dose. METHODS: A prospective randomized study included 524 adults--260 in the Bio-Hep group and 264 in the Engerix-B group. Both vaccines were administered in a three-dose regimen given at 0, 1 and 6 months, and adverse events were recorded on a diary card 5 days after each vaccination. Immunogenicity was tested by measuring anti hepatitis B surface antibody. RESULTS: One month after the third injection, 98% of the BioHep subjects were found to be seroprotected vs. 85.1% of the Engerix-B group. In addition, the geometric mean titers were 2,203 mIU/ml and 326 mIU/ml in the Bio-Hep-B and Engerix-B groups respectively. An immunogenic advantage of Bio Hep-B was suggested by the rapid onset of antibody response--66.5% were seroconverted one month after the first injection as compared to 19.3% in the Engerix-B group. No unexpected adverse events were observed, and the recorded events were mild in both groups. CONCLUSIONS: BioHep, a novel recombinant HBV vaccine containing S, Pre-S1 and Pre-S2 protein components, at a lower dose, is safe and more immunogenic than the conventional HBV vaccine that contains only S protein. PMID- 11411196 TI - Comparison of QT dispersion between primary coronary angioplasty and thrombolytic therapy for acute myocardial infarction. AB - BACKGROUND: Previous studies have documented that reduction in QT dispersion after thrombolytic treatment in acute myocardial infarction depends on reperfusion status as well as infarct site. Primary percutaneous transluminal coronary angioplasty as compared with thrombolytic therapy has been shown to result in higher patency rates of the infarct vessel. OBJECTIVES: To evaluate whether primary PTCA has a more favorable effect on reducing QT dispersion in patients with acute MI as compared to thrombolytic treatment. METHODS: The study population included 42 consecutive patients (33 men, mean age 58 +/- 11 years) with acute MI (24 anterior wall, 18 inferior wall) who were treated with primary PTCA (group A, n = 21) or thrombolytic therapy (group B, n = 21) at 3.9 +/- 2 hours after symptom onset. QT intervals were measured before and 24 hours after treatment. RESULTS: On the admission electrocardiogram, patients with anterior MI had higher values of QT and QTc dispersions than patients with inferior MI (52 +/ 9 vs. 36 +/- 9 msec, P < 0.05 and 61 +/- 4 vs. 56 +/- 4 msec, P = 0.002, respectively). There was a significant reduction in QT and QTc dispersions from admission to 24 hours in all patients (from 50 +/- 9 to 37 +/- 9 msec, P < 0.001 and from 59 +/- 5 to 42 +/- 5 msec, P < 0.001, respectively), and also in group A (from 49 +/- 8 to 32 +/- 5 msec, P < 0.001 and from 58 +/- 5 to 38 +/- 3 msec, P < 0.001, respectively) and in group B patients (from 51 +/- 10 to 42 +/- 10 msec, P < 0.01 and from 60 +/- 4 to 46 +/- 5 msec, P < 0.001, respectively). QT and QTc dispersions were found to be shorter in group A at 24 hours after treatment than in group B (32 +/- 5 vs. 42 +/- 10 msec, P < 0.001 and 38 +/- 3 vs. 46 +/- 5 msec, P < 0.001, respectively). CONCLUSIONS: Reperfusion therapy with primary PTCA or thrombolytic agents reduces QT and QTc dispersions in acute MI. QT and QTc dispersions after reperfusion treatment are shorter with primary PTCA than with thrombolytic therapy. PMID- 11411197 TI - Epidemiology of hepatitis B virus infection among family members of chronic carriers in Israel. AB - BACKGROUND: Hepatitis B is a major problem worldwide. Israel has intermediate endemicity for hepatitis B virus, and an annual carrier rate of 1-3%. OBJECTIVE: To evaluate both the prevalence of HBV infection among family members of HBV carriers and the competence of family practitioners in performing a comprehensive assessment. METHODS: A total of 152 HB surface antigen-positive blood donors were discovered in our subdistrict during the years 1993-97. Their family physicians were questioned regarding the patients' family members. Specific information on 85 spouses and 200 children was also obtained. RESULTS: Among the 85 married carriers, 5 of the spouses (5.9%) were found to be HBsAg positive. None of the 200 children was HBsAg positive. We found that in a third (n = 52) of the patients, the sexual partner had never been tested by a primary care physician. Patients were not routinely tested for HB e antigen or anti-HBe antibodies. Neither the parents nor the siblings had undergone any serological evaluation. However, most family members of the carriers had received an HBV vaccine from their family physicians. CONCLUSIONS: Our findings show that horizontal transmission of HBV among spouses of HBV carriers still exists. We did not find any vertical transmission, probably due to male predominance and previous vaccination. Family physicians should be trained to perform an extensive serological evaluation of family members of patients with chronic HBV infection, including parents and siblings, and should vaccinate seronegative family members. PMID- 11411198 TI - Do Ashkenazi Jews have a higher than expected cancer burden? Implications for cancer control prioritization efforts. AB - BACKGROUND: Recent genetic susceptibility findings in Jews of Eastern European descent, commonly called Ashkenazi Jews, have led to concerns that they may be stigmatized as being more cancer prone than other groups. OBJECTIVE: To examine the hypothesis that site-specific or all-cancer incidence and mortality rates are higher than expected in Ashkenazi Jews worldwide when compared with referent populations. METHODS: A MEDLINE search was performed using keywords "Jews," "cancer," "incidence" and "mortality" to identify studies directly relevant to the primary study question. RESULTS: Little evidence suggested that all-cancer incidence or mortality is higher in Ashkenazi Jews than in North American non Hispanic whites. Ashkenazi Jewish men appear to have relatively low cancer rates, which may be due to lower tobacco use. Colorectal cancer was shown to disproportionately overburden Ashkenazi Jews, who may also be at increased risk for ovarian, pancreatic and stomach cancer, and non-Hodgkin's lymphoma. Little evidence was found supporting an elevated risk of breast cancer in Ashkenazi Jewish women. Rates of lung, cervical, penile and prostate cancers appear low in this population. Rate disparities were generally attributed to lifestyle differences, particularly diet and tobacco use, rather than to genetic predisposition. CONCLUSIONS: Ashkenazi Jews do not appear to have a higher total cancer burden than comparable North American populations. Any cancer rate differentials in this group are more likely to be related to lifestyle and dietary factors than to genetics. However, colorectal cancer rates in Ashkenazi Jews may be the highest of any ethnic group in the world and cancer controllers should consider this when developing future screening, diagnostic and policy strategies. PMID- 11411199 TI - The changing epidemiology of viral hepatitis A in Israel. AB - BACKGROUND: Between 1970 and 1979, there was an increase in the incidence of viral hepatitis in Israel with a shift of peak incidence to an older age in the Jewish population, followed by a declining trend during the early 1980s. In July 1999 universal immunization of infants against hepatitis A was introduced. OBJECTIVE: To evaluate the chan-ges in the epidemiology of viral hepatitis A in Israel during the past decade. METHODS: Viral hepatitis is a notifiable disease in Israel and cases are reported to the regional health offices, which in turn provide summary reports to the Ministry of Health's Department of Epidemiology. The data in this study were derived from the summary reports and from results of seroprevalence studies. RESULTS: Following the increase in the incidence of reported viral hepatitis (mainly due to type A) between 1970 and 1979, the rates then stabilized and around 1984 began to decline until 1992. Since then there has been a slight increase. Whereas until 1987 the rates were consistently higher in the Jewish population, since then they are higher in the Arab population. The shift in the peak age-specific incidence from the 1-4 to the 5-9 year age group observed in the Jewish population around 1970 occurred 20 years later in the Arab population. The previously described seasonality is no longer evident. Recent seroprevalence studies indicate that by age 18 years only about 30-40% of the Jewish population have anti-hepatitis A antibodies. CONCLUSIONS: The decline in the incidence of hepatitis probably reflects the changing socioeconomic condition occurring at different times in the two major population groups. Since hepatitis A accounts for almost all the acute viral hepatitis in Israel, the universal vaccination of infants introduced in 1999 should substantially lower the morbidity within the next few years. PMID- 11411200 TI - Patient perception of involvement in medical care during labor and delivery. AB - BACKGROUND: Patients who feel involved in their treatment have better outcomes than those who do not. OBJECTIVE: To identify determinants of perceived patient involvement in obstetric care. METHODS: A retrospective study was undertaken in 1,452 (83%) of 1,750 women sampled in November 1995 from maternity wards of 14 general hospitals in Israel. A postal and telephone survey using a self administered questionnaire included the following variables: hospital (identity number), patients' age, self-reported complications, previous deliveries, education, ethnicity, and number of obstetric interventions performed and/or considered. The main outcome measured was the reported involvement in decisions for obstetric interventions. RESULTS: Reported full involvement varied from 72% for epidural analgesia to 13% for forceps/vacuum extraction. Factor analysis identified two dimensions of perceived involvement: one for "routine" interventions (enema, monitoring, IV line and episiotomy), which are performed in Israel mostly by midwives, and another for "special" interventions (forceps/vacuum extraction, epidural or other analgesia, and cesarian section) performed by physicians. Logistic regression identified hospitals, younger age, number of interventions, and Arab ethnicity as correlates of a perceived non involvement in decisions for "special" interventions. CONCLUSIONS: Clinical setting, age and ethnicity affected patient perception of involvement in decisions for obstetric interventions. PMID- 11411201 TI - Transmission of hepatitis B and C viruses--update. PMID- 11411202 TI - Viral dynamics in hepatitis C. AB - The hepatitis C virus is an enveloped positive-sense single-stranded RNA virus, which has been classified into 6 major genotypes and over 100 subtypes. HCV replicates mainly in the hepatocyte. Recently, infectious HCV cDNA clones have been generated. Despite evidence that innate and adaptative humoral and cellular immune responses are activated as part of an antiviral defense, HCV has a remarkable ability to establish persistent infection. The analysis of viral kinetics using mathematical modeling shows a relative steady state without treatment, while an immediate biphasic HCV decline occurs in blood during successful treatment, the latter being predictive of clearance of HCV by the end of treatment. PMID- 11411203 TI - Pancreas: a sex steroid-dependent tissue. AB - Sex steroid hormones (estrogens, progestagens and androgens) have been associated with healthy and neoplastic pancreatic biology, although the precise significance of the findings has not been well established. Receptors for the three different types of SSH are expressed in normal and tumoral pancreatic tissue with varying profiles related to cell origin (exocrine or endocrine), to type of neoplasm, and probably even to tumoral behavior. The activity of specific enzymes involved in the synthesis and transformation of SSH are increased in some neoplastic pancreatic tissues, which may influence the circulating concentrations of these hormones, such as the low serum testosterone:dihydrotestosterone ratio described in male patients with pancreatic carcinoma. Different patterns of age and gender related incidence and growth of neoplasms have been identified. Experimental studies have shown that pancreatic carcinogenesis is promoted or inhibited by SSH. At present, the data supporting hormonal manipulation for the treatment of these tumors are non-conclusive. Normal and tumoral pancreatic tissues may be regarded as a target for SSH and an additional site of biosynthesis. The influence of these hormones on physiological activities is not well known but should be further explored. The study of SSH in pancreatic neoplasms will provide clues about its origin, development, tumoral behavior, prognosis and more specific hormonal therapy. We review here the evidence favoring the role of SSH and their possible clinical implications in pancreatic function. PMID- 11411204 TI - Epidemiology of diabetes in Mexico and associated coronary risk factors. PMID- 11411205 TI - Main role of cytokines in autoimmunity. PMID- 11411206 TI - Early cancer in gastric Crohn's disease. PMID- 11411207 TI - The mosaic of autoimmunity. A classical case of inhalation of a polyclonal activating factor in a genetically and hormonally susceptible patient leading to multiple autoimmune diseases. PMID- 11411208 TI - Fatal liver necrosis associated with the use of nitrofurantoin. PMID- 11411209 TI - Acute encephalopathy preceding Shigella infection. PMID- 11411210 TI - Appendicitis in an elderly women: CT diagnosis. PMID- 11411211 TI - Intractable shoulder pain in a patient on chronic hemodialysis. PMID- 11411212 TI - Military medicine in the Crusaders' Kingdom of Jerusalem. PMID- 11411213 TI - [The advancement of G protein and coupled signal transduction pathways]. AB - G protein coupled signal transduction system is one major transmembrane signaling pathways in cell. Research of biochemistry and pharmacology in medicine showed that a lot of medicines play their role through G protein coupled signaling pathways. Thus, systematic study on G protein structure and functions had very important effects on new medicine producing. In the process of creature evolution, there are highly conserved amino-acid sequences in G protein. Study on G protein and coupled signal components in plants and insects signaling processes may benefit to determine the mechanism of crop resistance to disease and pest insect and resistant chemicals effect on insect behavior and physiology. This paper reviews the research progress and prospect of practical use in recent studies of G protein and coupled components in animal, insect and plant signaling pathways. PMID- 11411214 TI - [New advances and future perspectives in mammal cloning]. AB - To the great extent, the study and application of transgenic animal are restricted by the inherent limitation of pronuclear microinjection. Recently, the rapid progresses in gene targeting and cloning of somatic cells have shown that the combination of these two technologies will become a virtual way to producing large transgenic animals. PMID- 11411215 TI - [The production and scavenging of reactive oxygen species in plants]. AB - The imposition of environmental stress leads to increased production of reactive oxygen species (ROS) in plant cells, Which can damage proteins, membrane lipids, DNA and other cellular components. Plants have evolved enzymatic and no-enzymatic protection mechanisms that efficiently scavenge ROS. Enzymatic detoxication system includes superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and glutathione peroxidase (GAX); no-enzymatic antioxidants include ascorbic acid, glutathione, mannitol and carotenoids, which occur in high concentrations in plants. The over-expression and accumulation strategies of these antioxidants in plants have been followed up to now, and have gained many transgenic plants showing increased stress tolerance. PMID- 11411216 TI - [Molecular cloning and expression of the cDNA encoding angiogenesis inhibitor K4K5 with Pichia pastoris]. AB - Kringles of human plasminogen except kringle 4 can inhibit the endothelial cell growth. To determine whether recombinant plasminogen kringle 4-5 (r-K4K5) can inhibit the growth of bovine capillary endothelial (BCE) cell and the angiogenesis of chick embryo chorioallantoic membrane (CAM), we cloned and constructed the modified cDNA region encoding kringles 4 and 5 (K4K5) of human plasmin (ogen) and expressed it with pichia multi-copy expression system. One clone, with the most productive expression was selected from hundreds of transformants. Our data showed that the expression product r-K4K5 (MW 21.5 kD) amounted to 150-250 mg/liter, over 80% of the total secreted protein. It could, dose-dependently, inhibit BCE cell proliferation and inhibit chick embryo CAM angiogenesis. PMID- 11411217 TI - [Cloning and expressing of an anti-CD5 single chain antibody]. AB - Novel anti-CD5 single-chain fragment of variable domain (ScFv) was cloned and expressed. Anti-CD5 ScFv was constructed with cDNA fragments of heavy and light variable regions (VH and VL) which were reverse-translated from poly (A) mRNA of hybridoma cells producing anti-CD5 McAb. By phage displaying, ScFv in form of ScFv-g3p fusion protein was panned based on its binding capacity to the CD5 antigens on the cell surface of Molt-4 cells. Assayed by the cell-ELISA, 4 clones were found to have high affinity to CD5 antigen. DNA sequencing confirmed that the VH fragment is 339 base pairs, the VL 300 bp, and the ScFv belongs to the mice gene family. In E. coli HB2151 the soluble ScFv-CD5 was expressed mainly in the periplasm. PMID- 11411218 TI - [The expression of tPA directed by the bovine BLG regulatory elements in the mammary gland of transgenic mice]. AB - In order to get the regulatory elements which are essential for generating mammary gland bioreactors, the whole 8.4 kb bovine BLG gene was obtained by PCR amplification. The 1.6 kb chicken lysozyme matrix attachment region (MAR) was used to overcome position effects. The bovine BLG-tPA expression vector was constructed and the BLG-tPA fusion gene was introduced into fertilized eggs of mice by microinjection to generate transgenic mouse. 170 offsprings were obtained, of which 9 were proved to be transgenic mice based on PCR and Southern blot analysis. The tPA expression level amounted to 12 micrograms/mL in the milk of mice. The bovine BLG-tPA fusion gene integrated in the founders was inheritable. PMID- 11411219 TI - [Cloning of VEGF receptor KDR and its expression in insect cells]. AB - The cDNA fragment of the first 3 loops of VEGF receptor, KDR, was cloned by PCR and inserted into a baculovirus expression plasmid pFASTBACI. The competent E. coli DH10BAC cell, which contain another plasmid with baculovirus genome in it, was transformed with pFASTBACI-KDRn3. Homologous recombination in the prokaryotic cells resulted in a recombinant plasmid containing KDRn3 in baculovirus genome. Transfection of the insect cell SF-9 with above plasmid generated a recombinant baculorvirus contain target gene fragment. SDS-PAGE and Western blot analysis of the supernatant of the infected SF-9 cell showed that KDRn3 was secreted in the medium. The recombinant protein was verified with Western blot and tested for their binding activity with VEGF. Its anti-angiogenic activity was assayed on chorionic allantoic membrane(CAM) of fertilized egg. The results showed that the recombinant protein could inhibit new vessel formation on CAM of fertilized eggs. PMID- 11411220 TI - [Expression in Escherichia coli and immunogenicity of rotavirus VP7]. AB - Rotaviruses are the single most important cause of severe diarrhea in young children all over the world. VP7 is the major outer capsid and is a primary candidate for inclusion in a subunit or recombinant vaccine. Part of the VP7 gene containing all the three antigenic regions was expressed as a chimeric protein with glutathione S-transferase (GST) in E. coli. The chimeric protein, representing about 30% of the total protein of the recombinant-plasmid-carrying bacteria, reacted with polyclonal antibodies raised against whole virus. Immunization of sero-negative rabbits and mice with purified fusion-protein generated both virus-binding and neutralizing antibodies. PMID- 11411221 TI - [Enzymatic conversion of cephalosporin C to glutaryl-7-aminocephalosporanic acid using whole cells of the yeast Trigonopsis variabilis FA10]. AB - A process for the production of glutaryl-7-aminocephalosporanic acid (GL-7ACA) from cephalosporin C(CPC) using permeabilized cells of yeast Trigonopsis variabilis FA10 containing D-amino acid oxidase (DAO) is described. It was found that the bioconversion of CPC to GL-7ACA was interfered by the catalase activity presented in the cells that hydrolyzed the hydrogen peroxide and resulted in the accumulation of alpha-keto-adipyl-7-ACA (AKA-7ACA) and decrease of GL-7ACA yield. the methods to overcome this problem including the addition of extra H2O2 and use of catalase inhibitor, NaN3, were developed and the rate of GL-7ACA from CPC were 73% and 70.1%, respectively. Another alternative method was to incubate the permeabilized FA10 cells at pH10.5-11.0 for 30 minutes at 20 degrees C which served to selectively inactivate the catalase. In the bioconversion of CPC to GL 7ACA using pH10.5-treated cells without catalase activity, the high reaction yield of GL-7ACA(84%) was achieved. PMID- 11411222 TI - [Technical improvements in genetic manipulation of Pichia pastoris and their application in hirudin expression]. AB - Pichia pastoris has become an increasingly popular host for heterologous protein production. However, there is neither a high-efficient transformation method nor a fast colony-PCR assay for the yeast yet. In this paper, we report a transformation procedure by electroporation, which reaches the value of up to 2800 transformants/microgram DNA. By using a cold and heat treatment and a modified PCR buffer, we established a simple and reliable colony-PCR protocol to detect recombinant P. pastoris clones, which is comparable to the conventional assay for E. coli colonies. With these two novel techniques, we have successfully achieved the expression of hirudin, an antithrombin agent, in Pichia pastoris. The secreted hirudin maintains a biological activity of 82 antithrombian units per milliliter supernatant from the media. PMID- 11411223 TI - [Expression of delta 6-fatty acid desaturase gene from Mortierella alpina in Saccharomyces cerevisiae]. AB - delta 6-fatty acid desaturase is the rate-limiting enzyme of the desaturation of linoleic acid in the production of an essential fatty acid, gamma-linolenic acid. The 1.4 kb fragment in plasmid pTMACL6 encoding delta 6-fatty acid desaturase from Mortieralla alpina ATCC16266 was subcloned into the yeast-E. coli shuttle vector pYES2.0, thus an expression recombinant plasmid pYMAD6 containing target gene was constructed and obtained in the SC-Ura media. The pYMAD6 was introduced into defective mutant INCSc1 of Saccharomyces cerevisiae by LiAc method. When linoleic acid was provided as an exogenous substrate to the yeast cultures expressing delta 6-fatty acid desaturase activity under appropriate media and temperature condition, the level of gamma-linolenic acid reached 31.6% of the total yeast fatty acids by GC-MS detecting, which is the highest report of delta 6-fatty-acid desaturase gene in S. cerevisiae. PMID- 11411224 TI - [Construction of yeast fusants of directly transform starch into ethanol]. AB - To obtain strains that are able to efficiently produce ethanol directly from starchy material, we have used the protoplast fusion technique to construct hybrids between Saccharomyces cerevisiae and Candida tropicalis. The isolation of fusants used a selective system by a irreversible biochemical inhibitor Iodoacetic Acid(ID), and the fusion frequency was 9.2 x 10(-6). The cell size and DNA content of the fusants were determined. Through testing some productive properties such as specific growth rate, saccharification ability, ethanol tolerance and the activities of alpha-amylase, saccharogen amylase, we selected two fusants--F-1 and F-5. They were more superior and their ethanol production came up to 8.8% and 11.5%. PMID- 11411225 TI - [Construction of luciferase reporter plasmid which is under the control of dioxin responsive enhancers]. AB - To improve the bio-detection of dioxin-type chemicals, we constructed a plasmid containing reporter under the control of dioxin-responsive enhancer. After connected the enhancer from the plasmid pHAV with the MMTV promoter of the plasmid pCatM, the connected segment was linked with the luciferase vector. Then the recombinant plasmid was transfected to the HepG2 human hepatoma cell line, and the luciferase activity was induced by 2,3,7,8-TCDD. The result suggested that expression of the recombinant luciferase was controlled by the dioxin responsive enhancer, and there were a linear relationship between the activity of luciferase and certain range of 2,3,7,8-TCDD concentration. It is promising that the transfected cell line could be used for rapid screen and semi quantitation of dioxin-type chemicals, and a standard and routine method for the detection of dioxin-type chemicals by this recombinant plasmid is being under way. PMID- 11411226 TI - [Purification and biological activity of rh-leptin expressed in Escherichia coli]. AB - The human leptin was successfully expressed with high level in E. coli under the control of PL promotor. The yield of recombinant protein was over 40% of total cellular protein and expressed as inclusion bodies. The recombinant human leptin (rh-leptin) was purified with gel filtration, anion-exchange and reverse chromatography. Refolding was achieved by gradually reducing denaturant using a diafiltration method. The refolded rh-leptin was characterized by SDS-PAGE, Western-blotting and its first 15 amino acid residues sequence of the N-terminal. The purified product was found to be biologically active, reducing the food intake and body weight gain upon testing in BALB/c mice. PMID- 11411227 TI - [The construction and evaluation of antisense RNA expression vector targeting at HCV 5'NCR]. AB - Antisense RNA is an important field of antisense technique. In order to explore a novel approach for the treatment of hepatitis C virus (HCV) infection and prove the availability of transgenic cellular model HepG2. 9706, an antisense RNA was designed targeting at the highly conserved 5'NCR and translation initiation site of HCV RNA at nt positions 13-397. It was inserted into the downstream of SV40 promotor sequence of pGL3 control vector in which luciferase gene is deleted. The antisense RNA expression vector (pHCV-asR) was constructed and identified by PCR, endonucleases reaction and DNA sequencing. Its expression in transfected HepG2 cells was tested by RT-PCR. To evaluate the inhibitory activities of pHCV-asR, HepG2. 9706 cells were transfected using this construct via Lipofectin method. Luciferase activity in cell lysates was measured for quantitative determining antiviral effects within the cells. The results showed that the inserted sequence of the pHCV-asR is the same as the designed sequence and can express in HepG2 cells. It was also found that pHCV-asR in HepG2. 9706 has a dose-dependent inhibitory effects on luciferase expression controlled by HCV 5'NCR with a inhibitory rate of 57%. PMID- 11411228 TI - [Cloning, high level expression and purification of porcine IFN gamma]. AB - Peripheral blood lymphocytes from a single swine were stimulated with Concavadin A for 17 h, and the total RNA was isolated from it. Then, the mRNA specific for porcine IFN gamma was amplified by reverse transcription polymerase chain reaction. After sequencing, the IFN gamma gene has been successfully inserted into vector pJLA-503 and highly expressed in E. coli. Recombinant porcine IFN gamma expressed as inclusion body, which was dissolved in 7 mol/L guanidine chloride and subsequently renatured by dilution in refolding buffer containing 0.5 mol/L L-arginine. In order to obtain pure protein, the renatured IFN gamma was purified by the chromatographies of SP-Sepharose FF and Sephacryl S-200 HR. As a result, the final pure product can been seen as a single band in SDS-PAGE, and the cytokine activity was verified by inhibiting the cytopathic effect. PMID- 11411229 TI - [Culture medium optimization and primary kinetics analysis for nisin production]. AB - Response surface methodology was used to optimize a medium for nisin production of Lactococcus lactis. In the first optimization step the influence of sucrose, soybean peptone, yeast extract, potassium dihydrogen phosphate, sodium chloride, and magnesium sulfur on nisin production was evaluated using a fractional factorial design. Potassium dihydrogen phosphate influenced nisin production positively while soybean peptone affected nisin production negatively. The other components had no significant effect on nisin production. The path of steepest ascent was used to approach the optimal region of the medium composition. In the third step the optimal concentrations of KH2PO4 and soybean peptone were determined by a central composite design and response surface analysis. The optimized medium allowed nisin production to be increased from 1074 IU/mL to 2150 IU/mL. The kinetic analysis showed that nisin production fashion at optimized and non-optimized media was not changed and maintained partially growth-associated. But the specific growth rates and the specific nisin production rates for the strain at the optimized medium were bigger than the ones at the non-optimized medium after the cells entered the middle of exponential phase. PMID- 11411230 TI - [Kinetics of anaerobic ammonia oxidation]. AB - The kinetic characteristics of anaerobic ammonia oxidation was studied. The cell yield was 1.573 mgVS (mmolNH4+)-1 and the cell decay coefficient was 0.052 mgVS(gVS.d)-1. The maximal rate of anaerobic ammonia oxidation and nitrite conversion rate by the mixed culture were 1.320-2.761 mmol(gVS.d)-1 and 14.497 mmol(gVS.d)-1, respectively. Km for ammonia and for nitrite were 1.801-4.215 and 0.468 mmol.L-1. Ki for ammonia was 38.018-98.465 mmol.L-1, the optimal ammonia concentration was 16.656 mmol.L-1. Ki for nitrite was 5.401-11.995 mmol.L-1. The optimal pH was around 7.5 and the theoretical value was 7.605. The optimal temperature was about 30 degrees C. The activation energy for Vmaxa, Kma, Kia and Kin was 37.316, 30.239, 33.695 and 30.473 kJ.mol-1. PMID- 11411231 TI - [Cloning and high level expression of mosquito detoxifying gene]. AB - The full-length cDNA of mosquito esterase B1 had been isolated, and subcloned into pBV220. The recombinant vector pBV220B1 was constructed and transformed into E. coli DH5 alpha. A 60 kD protein was induced by 42 degrees C and its expression was temperature-dependent. After 6 h induction, the target protein occupied 50% of the total protein. The expressed product existed in both inclusion body and soluble proteins in the cells. The amount of the soluble detoxifying enzyme increased along with the induction time. The data of detoxifying experiments indicated that the detoxifying enzyme in expression strain of E. coli can detoxified toxicity of organophosphate insecticides, it showed a clear detoxifying affect on hens poisoned by organophosphate insecticides. PMID- 11411232 TI - [Cloning, sequence analysis and high-level expression in Escherichia coli and activity assay of pac-1 gene from Schizosaccharmyces pombe]. AB - The Schizosaccharmyces pombe pac-1 gene product is a kind of dsRNA dependent ribonuclease, which has potential to degrade the dsRNA viral genome, the replication form of ssRNA viral genome and viroid genome. Therefore, to introduce the pac-1 gene into plants conferring them resistance to viruses is a new method of establishing the anti-virus transgenic plant. The pac-1 gene from the S. pmobe genome DNA isolated from China was cloned by means of PCR amplification. The pac 1 gene was inserted into the cloning vector pGEM-7Zf(+) by using restriction endonuclease Kpn I/BamHI. Sequencing analysis shows that it is a complete gene with 1095 necleotides. Compared to the reported pac-1 gene, its homology is significant, but with 5 nucleotides differences, leading to only one amino acid difference. Pac-1 gene was inserted into the prodaryotic expression vector pET 21(a) by using the restriction endonuclase Nde I/BamHI. It was induced by the IPTG in E. coli BL21 harbouring the recombinant vector pET-pac-1. The pac-1 gene product is analyzed by the SDS-PAGE. The result shows the product of pac-1 gene exists in the supernatant part as soluble form and in the precipitant part as inclusion bodies after the cells were lysed by ultrasonic wave. The supernatant was applied to detect the enzyme activity of pac-1 gene product. We concluded that pac-1 gene has the biological activity of degrading the CMV-dsRNA. PMID- 11411233 TI - [Cloning and expression of a synthetic gene encoding magainin-melittin hybrid peptide in Escherichia coli and studies on its antibacterial activity]. AB - A hybrid peptide gene was designed and synthesized. Its encoding peptide is constructed from residues 3-14 of magainin and residues 1-13 of melittin. The MA E gene was cloned into plasmids pUC18 and pBV220. By DNA sequencing, the whole sequences of this gene is confirmed to be correct. The recombinant plasmid pBMA-E was expressed in E. coli DH5 alpha. A gene product band can be seen with Tricine SDS-PAGE. The MA-E hybrid peptide was purified by immobilized metal affinity chromatography. Bioactivity assay was carried out in liquid turbidity method. The bactericide value to E. coli K12D31 is 0.182. PMID- 11411234 TI - [Purification and immobilization of the proteinase from mung bean burgeon inactivating soybean trypsin inhibitor]. AB - By 30%-60% s(NH4)2SO4 fractional precipitation, anion-exchange chromatographs on DEAE-Sepharose CL-6B, gel filtration on Sephacryl S-200 and anion-exchange chromatographs on Waters AP-1 column(ProteinTm-Pak DEAE 15HR), a proteinase which can inactivated STI was purified from mung bean(Phaseolus aureus) burgeon. It was stable at temperatures lower than 50 degrees C and pH7.5-8.5, and the Km and Vmax of the proteinase for STI was 769.2 alpha-N-benzoyl-L-arginine ethyl ester(BAEE)/mL and 115.3BAEE/min/mL respectively. The molecular weight of the proteinase was estimated to be 29.8kD by SDS-PAGE. The proteinase immobilized by polyacrylamide was stable at temperatures lower than 60 degrees C and pH7.0-9.0, and the apparent Km* and Vmax* of the immobilized proteinase for STI was 1303.8 (BAEE)/mL and 94.34(BAEE)/min/mL respectively. The half-life of the immobilized proteinase was about 12 days at 4 degrees C. PMID- 11411235 TI - [Effect of dissolved oxygen on the suspension culture of Taxus chinensis]. AB - The effect of dissolved oxygen controlled level on the taxol production by suspended cell culture of Taxus chinensis was examined in shake flasks and in a 20L aerated and agitated bioreactor with a six-flat-bladed turbine. It was found in shake flasks that oxygen supply exhibited significant influence on the production of taxol. Further investigation in bioreactor showed that noly 1.5 mg/L taxol was produced when DO(dissolved oxygen) level was 20%. Taxol production increased when DO level was increased to 40%-60%. Moreover, taxol accumulation was remarkably improved by the cultivation of cells initially at the DO level of 60% during the growth phase for 14 days followed by changing the DO level to 20% 40% during the taxol production phase for another 10 days. An optimum taxol level of 7.2 mg/L was obtained by the dissolved oxygen controlled process. PMID- 11411236 TI - [Study on fermentation of long-chain dicarboxylic acid based on enzymology]. AB - In this paper, we studied the effects of the adding time of alkane on the expression of P450 and production of dicarboxylic acid. A novel fermentation process, in which no or a little alkane was added to make the cells growing more quickly during the growth stage, followed by the addition of alkane to induce cytochromes P450 for 6-8 hours, was established. The results showed that the new process was much better than the old process on inductivities of cytochromes P450 and production of dicarboxylic acid. The new process improved nearly 14.56% expression of P450 and 14.15% production of dicarboxylic acid. PMID- 11411237 TI - [Studies on eicosapentaenoic acid production by submersion culture of Cryphecodinium cohnii]. AB - The effects of the incubation temperature, initial pH of the medium, carbon source and nitrogen source on the production of Crypthecodinium cohnii C98 were studied. Through orthogonal experiments, the optimum culture medium was obtained (g/L): Glucose, 65; Yeast extract, 2.0; KNO3 3.0, KH2PO4 1.0, MgSO4.7H2O 0.6, NaCl 5.8, CaCl2.2H2O 0.1, ZnSO4.7H2O 0.0067, FeCl3.6H2O 0.014, CuSO4.5H2O 0.0004, MnSO4.H2O 0.0002. Under the optimum culture conditions, the microalgae dry cell weight and eicosapentaenoic acid was 18.3 g/L and 0.891 g/L, respectively. The submersion culture process was analysed. PMID- 11411238 TI - [Esterification of octanoic acid with 1-octanol catalyzed by lipase in W/O microemulsions and in microemulsion-based organogels]. AB - The esterification reactions of octanoic acid with 1-octanol catalyzed by Candida lypolytical (CL) lipase was studied in water-in-oil microemulsions formed by water/bis-(2-ethylhexyl)sulfosuccinate sodium (AOT)/isooctane. The results of kinetic study showed that the reaction follows a Ping-Pong Bi-Bi mechanism. The values of apparent kinetic parameters were determined. Lipase has also been immobilized in gelatin-containing AOT microemulsion-based organogels (MBGs) for retention of catalytic activity. These lipase-containing MBGs proved to be a solid-phase catalysts for use in apolar organic solvents, retaining its higher activity after many runs of esterification reactions. PMID- 11411239 TI - Cardiopulmonary monitoring at home: the CHIME monitor. AB - A new physiologic monitor for use in the home has been developed and used for the Collaborative Home Infant Monitor Evaluation (CHIME). This monitor measures infant breathing by respiratory inductance plethysmography and transthoracic impedance; infant electrocardiogram, heart rate and R-R interval; haemoglobin O2 saturation of arterial blood at the periphery and sleep position. Monitor signals from a representative sample of 24 subjects from the CHIME database were of sufficient quality to be clinically interpreted 91.7% of the time for the respiratory inductance plethysmograph, 100% for the ECG, 99.7% for the heart rate and 87% for the 16 subjects of the 24 who used the pulse oximeter. The monitor detected breaths with a sensitivity of 96% and a specificity of 65% compared to human scorers. It detected all clinically significant bradycardias but identified an additional 737 events where a human scorer did not detect bradycardia. The monitor was considered to be superior to conventional monitors and, therefore, suitable for the successful conduct of the CHIME study. PMID- 11411240 TI - Noise sensitivity of three surface ECG fibrillation detection algorithms. AB - The widening application of automatic external defibrillators (AEDs) presents very strong requirements for external electrocardiogram (ECG) signal analysis. Highly accurate detection of shockable rhythms is required, aimed to approach the maximum of 100% sensitivity and specificity. In a previous study the performance of five well known detection algorithms was assessed by test signals from the ECG signal databases of the American Heart Association (AHA) and the Massachusetts Institute of Technology (MIT). The results obtained were used as a basis for testing the noise sensitivity of three of these algorithms. Realistic noise was obtained by simulation and recording of signal disturbance by various motions during resuscitation and defibrillation episodes (body shudder convulsions and gasps, cable movement, car transportation). The sensitivity and specificity of the detection algorithms were evaluated using electrocardiogram signals mixed with these noises. PMID- 11411241 TI - Assessment of computer-controlled inflation/deflation for determining the properties of PTCA balloon catheters with pressure-volume curves. AB - Restenosis is a significant problem after percutaneous transluminal coronary angioplasty (PTCA), leading to further angina or even death. The mechanisms causing restenosis are not well understood and the effect of the angioplasty balloon on the artery wall is generally unknown. The aim of the research presented here was to assess the repeatability with which measurements of balloon pressure-volume characteristics could be made, to ensure that different balloons could be distinguished easily, an essential first step before the arteries themselves could be characterized. An inflation/deflation device was inflated with and without an attached balloon under computer control at a constant speed to 8 bar. The accuracy and repeatability of the technique was assessed within a single measurement session and between measurement sessions on six separate days. For each measurement session, four repeated inflations/deflations were carried out and the average was determined. Compliance (volume change/pressure change), inflation volume and pressure drop at 8 bar were determined for various balloon catheters of different size, type and manufacturer. The system repeatability SD for measuring volume without a balloon was +/- 0.0005 ml within a single measurement session, and +/- 0.002 ml between independent days. With an attached balloon the repeatability SD for measuring volume was +/- 0.001 ml within, and +/ 0.003 ml between days. Compliance was determined with a repeatability SD of +/- 0.16 microliter bar-1 within, and +/- 0.94 microliter bar-1 between days. The inflation/deflation device and technique employed allowed small changes in volume and compliance to be differentiated. The system also enabled differences due to the material characteristics for different PTCA balloons to be determined. Measured balloon characteristics were similar to the data given by manufacturers. This gives confidence in development of the technique for clinical use. PMID- 11411242 TI - Defibrillation and the geometry of the heart: a novel measurement with implications for defibrillation mechanisms. AB - We present a novel measurement for studying defibrillation mechanisms: the time course of changes in the size of the left ventricular (LV) cavity within 500 ms following defibrillation. Mechanical changes can be linked to electrical mechanisms via an understanding of excitation-contraction coupling. Eight mongrel dogs were internally defibrillated 5-50 seconds (including backup shocks) after the onset of 20 ventricular fibrillation (VF) episodes per animal. Two dimensional, short axis, LV cavity, ultrasound images were recorded at 30 frames per second just prior to inducing VF, during defibrillation and following the shock. Each frame was individually analysed to yield the LV cavity area as a function of time. Defibrillation shocks were followed by a highly reproducible phenomenon: (1) a dramatic and rapid increase in LV area, (2) a more or less prominent LV area plateau and (3) a decrease in the LV area. The peak fractional area increase ranged from 1.65 to 4.64 times larger than the baseline (LV area just prior to defibrillation), averaging 2.18 +/- 0.686. Successful shocks took significantly longer (p < 0.01) to return to 1.3 times the baseline (407 +/- 209 ms) than unsuccessful shocks (296 +/- 130 ms). Extrapolating to electrical mechanisms, our novel measurement demonstrates that defibrillation causes immediate relaxation and therefore suggests a significant role for deexcitation in defibrillation. PMID- 11411243 TI - An analogue instrument for the measurement of respiratory impedance using the forced oscillation technique. AB - The successful development of non-invasive assisted ventilation via a nasal mask has led to the requirement for improved methods of monitoring the respiratory system. Measurements of respiratory impedance during assisted ventilation using acoustic methods are showing considerable promise in providing this clinical data. The method involves imposing very low amplitude acoustic oscillations onto the spontaneous or assisted respiratory airflow. The subsequent interaction with the respiratory system modifies the pressure and flow relationship of the forced oscillations. The ratio of pressure to flow can be used to measure in real time the absolute value of respiratory input impedance. As a result a versatile, non invasive and repeatable profile of respiratory mechanics is obtained. Using a 5 Hz conventional forced oscillation frequency, data highly sensitive to airflow obstruction and capable of tracking mechanical events and responses become available to the clinician. Recent studies with ventilated and sleeping subjects confirm future potential for the forced oscillation technique in vivo. We describe an instrument named the respiratory impedance monitor (5 Hz), which utilizes the forced oscillation technique (FOT). This fully developed device is designed in accordance with agreed international clinical guidelines. Comparative clinical data from normal subjects and patients with chronic obstructive pulmonary disease is presented. PMID- 11411244 TI - Pulsatile electrical impedance response from cerebrally dead adult patients is not a reliable tool for detecting cerebral perfusion changes. AB - The original objective of this work was to verify the possibility of using electrical pulsatile cerebral impedance measurements as a diagnostic aid for assessing the brain-death condition in adults; a subordinate target was to validate a simple method for detecting perfusional changes in the brain. To this end, impedance signals were recorded, for a comparative study, from both live subjects and brain-dead patients, using a simple four-electrode arrangement. Rather unexpectedly, pulsatile transcephalic impedance waveforms exhibiting a temporal dependance similar to those of live subjects were detected in artificially ventilated, cerebrally dead, adult subjects; distributions of the time delays between impedance peaks and ECG peaks were also recorded for the two groups (dead and live subjects). These data provided no evidence, at the 1% significance level, against the hypothesis that the two sample groups are drawn from identical populations. The detection of impedance variations from brain-dead patients can be explained by the residual persistence of blood flow through the scalp, by mechanical variations synchronous with the heart beat and by the presence of the oscillating flow and the systolic spikes that precede the final blood flow arrest. The fact that impedance variations can be traced back to a multiplicity of causes, unrelated to the normal unidirectional flow, renders the transcephalic impedance method inappropriate for detecting cerebral perfusion changes in adults. This conclusion is also strengthened by some theoretical results recently derived from a multilayer model of the head. PMID- 11411245 TI - Measurement of intercompartmental fluid shifts during haemodialysis in children. AB - Seven children (age range 12-19 years, post-dialysis weights 23-43 kg) were studied during 20 haemodialysis sessions. Impedance between wrist and ankle (on the non-fistula side) was recorded using the Xitron 4000B analyser. A 2 ml sample of blood was taken for total protein and haematocrit from the arterial line at the start of dialysis. At approximately 20 minute intervals during dialysis, the time and volume of ultrafiltrate removed were recorded, and a simultaneous measurement of whole body impedance made over 25 logarithmically spaced frequencies in the range 5-500 kHz. A 2 ml sample of blood was also taken, from which serum protein and haematocrit were calculated. Hypotensive episodes occurred during four haemodialysis sessions. The percentage change in extracellular fluid (ECF) volume was calculated, at each sample time for each session, using the impedance measurements and ultrafiltration measurements (denoted delta Vi and delta U respectively). Changes in the intravascular volume were estimated using measurements of haematocrit and serum protein (and denoted delta Vh and delta Vp respectively). Least-squares regression gave delta Vi = 3.77 delta Vh, 1.33 delta Vp and 0.39 delta U, and r2 = 0.72, 0.94 and 0.95 respectively (p < 0.0001 in each case) for the 16 dialysis sessions without hypotensive episodes. Similar analysis of four dialysis sessions with hypotensive episodes gave similar relationships with correlation coefficients 0.64, 0.92 and 0.94. These relationships may not be accounted for by the anthropometric terms alone in the impedance equations. Impedance measurements also detected the addition of 300 ml isotonic saline given at the onset of each of the four hypovolaemic episodes. The regression equations support the following hypothesis: during haemodialysis, ultrafiltrate is removed from the intravascular volume but is replenished by fluid from the interstitial volume. The reduction in ECF volume measured by impedance (where the ECF comprises the intravascular and interstitial volumes) delta Vi is therefore greater than delta Vh and delta Vp, which only measure intravascular volume, but less than delta U since the ECF is replenished by fluid from the interstitial space. That delta Vh is greater than delta Vp may be due to protein loss during dialysis. The results suggest that whole body impedance measurements reflect changing body water distribution during dialysis in children. PMID- 11411246 TI - Electrical resistivity of the upper arm and leg yields good estimates of whole body fat. AB - Single frequency bioelectrical impedance analysis is an inexpensive, quick and painless means of estimating body composition. However, current approaches to estimating body composition from segment resistivity have some drawbacks. The purpose of this study was to overcome these limitations by developing a better model of the relationship between resistivity and body composition. A three compartment model of body mass and body segment resistivity is presented and calibrated to predict %Fat estimated by underwater weighing. The subject population from which the data were obtained was heterogeneous. Both sexes were represented, as were a range of ages (21-44 years), ethnic backgrounds, body masses (47.0-129.0 kg) and body compositions (%Fat = 8.7-50.7%). Based on resistivity measurements from the upper arm and upper leg, and measurements of subject height and weight, the model predicted %Fat with errors comparable to those reported for other methods based on segmental resistivity. All the terms in the calibrated model represented a physical component of the body and show reasonable agreement with resistivity measured in tissue samples. In short, predicting %Fat from resistivity of the proximal arm and leg segments compares favourably with other methods based on segment resistivity. PMID- 11411247 TI - Prenatal diagnosis of QT prolongation by fetal magnetocardiogram--use of QRS and T-wave current-arrow maps. AB - To determine the T wave of a fetal magnetocardiogram (FMCG), we have evaluated the T/QRS ratio and obtained current-arrow maps that indicate weak currents. We measured FMCG signals for 52 normal fetuses and two abnormal fetuses with prolonged QT waves by using three superconducting quantum interference device (SQUID) systems: a nine-channel system, a 12-channel vector system and a 64 channel system. The T/QRS ratio was calculated for all the normal fetuses from the maximum magnitudes of the QRS complex and the T wave. Current-arrow maps of the QRS complex (R wave) and T wave were obtained by using the 64-channel system, and the phase differences of the total-current vectors were calculated by using the current-arrow maps. The results showed that the T/QRS ratio had a wide variability of 0.35 for the normal fetuses. However, the magnitude of the prolonged T wave was as weak as the detection limit of the SQUID magnetometer. Although the T/QRS ratios for the fetuses with QT prolongation were within the normal range (< 0.35), the weak magnitude of the prolonged T wave could be evaluated. On the other hand, by comparing the current-arrow maps of the R and T waves for the normal fetuses, we found that the maximum-current arrows were indicated as either in the same direction or in opposite directions. These patterns could be identified clearly by the phase differences. Very weak prolonged T waves for the two abnormal fetuses could be determined by using these current-arrow maps and phase differences. Consequently, although the T/QRS ratios of FMCG signals have a wide distribution, we have concluded that the current arrow map and phase difference can be used to determine the T wave of an FMCG signal. PMID- 11411248 TI - Photoplethysmographic measurement of changes in total and pulsatile tissue blood volume, following sympathetic blockade. AB - Epidural anaesthesia, used for pain relief, is based on blocking the sensory and the sympathetic nerves in the lower part of the body. Since the sympathetic nervous system regulates blood vessel diameter, the sympathetic block is also associated with several haemodynamic changes. In the current study photoplethysmography (PPG) was measured on toes and fingers of patients undergoing epidural anaesthesia. Three parameters, which are related to the change in total and pulsatile tissue blood volume, were derived from the PPG baseline and amplitude. All parameters showed statistically significant increase in the toes after the sympathetic block, indicating higher arterial and venous blood volume and higher pulsatile increase in the arterial blood volume (higher arterial compliance) in the toe. These haemodynamic changes originate from the lower tonus of the arterial and venous wall muscles after the sympathetic block. In the fingers the PPG parameters based on the change in PPG amplitude decreased after the sympathetic block, indicating lower compliance. The measurement of the haemodynamic changes by PPG enables the assessment of the depth of anaesthesia, and can help control the adverse effects of the blockade on the vascular system. PMID- 11411249 TI - A fuzzy system for detecting distorted plethysmogram pulses in neonates and paediatric patients. AB - Pulse oximetry is a useful, quick, non-invasive and widely used technology for monitoring oxygen saturation (SaO2) for neonates and paediatric patients. However, pulse oximetry is fraught with artefacts, causing false alarms resulting from patient or probe movement. The shape of the plethysmogram is a useful visual indicator for determining the reliability of SaO2 numerical readings. If certain features could be defined that tag valid plethysmogram pulses, then automatic recognition of valid SaO2 values can be attained. We observed that the systolic upstroke time (t1), the diastolic time (t2) and heart rate (HR) extracted from the plethysmogram pulse constitute features which can be used for detecting normal and distorted plethysmogram pulses. Therefore, we developed a knowledge based system using fuzzy logic for classifying plethysmogram pulses into two categories: valid and artefact. A total of 22,497 pulse waveforms were used to define the system parameters. These were obtained from 13 patients with heart rates ranging between 62 and 209 beats min-1. A further 1420 waveforms obtained from another four patients were used for testing the system, and visually classified into 833 (59%) valid and 587 (41%) distorted segments. The system was able to classify 679 (82%) valid segments and 543 (93%) distorted segments correctly. The calculations of the system's performance showed 82% sensitivity, 86% accuracy and 93% specificity. We, therefore, conclude that the algorithm used in this system can be implemented in its present from for real-time SaO2 monitoring in intensive care for detecting valid and distorted plethysmogram pulses. PMID- 11411250 TI - Are ejection fractions from gated SPECT perfusion studies clinically useful? A comparison with radionuclide ventriculography. AB - Gated SPECT (GSPECT) was evaluated for the measurement of left ventricular ejection fraction (LVEF) by comparing with equilibrium gated radionuclide ventriculography (RNVG). A total of 99 subjects underwent GSPECT and RNVG imaging. All studies were acquired in list mode with GSPECT studies processed to give 16- and 8-frames per R-R interval, and RNVG studies 24 frames per R-R interval. The Cedars-Sinai QGS software was used to calculate ejection fraction from GSPECT studies. RNVG studies were processed using a manually drawn single region of interest technique. Comparison of LVEF from GSPECT with RNVG yielded correlation coefficients of 0.82 and 0.81 for 16- and 8-frame GSPECT studies respectively. The mean 95% prediction interval was 33 +/- 11 percentage points for both 16- and 8-frame studies, indicating a great disparity between predicted ejection fraction values from GSPECT and actual RNVG values. Subgroup analysis of 29 patients with pathological Q-wave evidence of myocardial infarction demonstrated a poorer correlation coefficient of r = 0.69. Subgroup analysis of 32 patients with end-diastolic volumes < 100 ml demonstrated a poorer correlation coefficient of r = 0.32. Ejection fractions calculated from 16- and 8-frame studies showed a correlation of 0.99 with a mean 95% prediction interval of 8.7 +/- 0.04 percentage points. The 8-frame studies underestimated LVEF by 3.6 +/- 2.3% compared to the 16-frame studies. In conclusion, left ventricular ejection fractions calculated using the QGS algorithm from GSPECT studies are inadequate for use in clinical practice. PMID- 11411251 TI - An electronic simulator for testing infant apnoea monitors that uses actual physiologic data. AB - An electronic simulator of physiologic signals used in infant monitoring has been designed, constructed and applied in the Collaborative Home Infant Monitor Evaluation (CHIME). A unique feature of the simulator is that it contains actual physiologic waveforms recorded from infants rather than artificial, idealized signals. The simulator stores breathing waveforms that can be used to test transthoracic-impedance- and inductance-plethysmography-based monitors, and heart rate channels are tested by playing a neonatal QRS complex at preset fixed rates or a variable rate as determined from infant recordings. The transfer characteristics of the simulator are constant over frequencies ranging from 0.5 to 8 Hz for the respiration channels. Data stored in memory are divided into 60 second epochs that can be presented to the monitor being tested in a programmable sequence. A group of 66 CHIME monitors was tested using a simulator programmed with 17 apnoea and bradycardia waveforms. The agreement between monitors as to the duration of detected apnoea decreases as the amount of artefact in the signal increases. Discrepancies between monitors in detecting apnoea duration were found to be similar to inconsistencies between CHIME investigators manually scoring similar waveforms. PMID- 11411252 TI - Recording of short-term finger blood pressure changes induced by an arterial occlusive thigh cuff: comparison between the modified oscillometric and Finapres techniques. AB - Blood pressure recordings obtained by the volume clamp method (Finapres monitor, Ohmeda, USA) were compared with measurements by using a novel modification of the oscillometric method (UT9201 beat-to-beat monitor, University of Tartu, Estonia). Step changes in total peripheral resistance were introduced in 11 healthy volunteers by rapid inflation to the suprasystolic pressure of bilateral thigh cuffs during a rhythmical exercise with the quadriceps muscles and simultaneously performed handgrip compression. The provoked short-term changes in mean arterial pressure (MAP) were similarly tracked by both devices. The agreement between the methods was better for MAP rise than drop. The group-averaged difference UT9201 minus Finapres did not exceed 1.2 mmHg (NS, p = 0.46) after cuff successive inflation but yielded 3.8 mmHg (significant, p < 0.05) after cuff successive deflation. PMID- 11411253 TI - Data logging technology in ambulatory medical instrumentation. AB - This paper reviews the advancements made in ambulatory data logging used in the study of human subjects since the inception of the analogue tape based data logger in the 1960s. Research into the area of ambulatory monitoring has been rejuvenated due to the development of novel storage technologies during the 1990s. Data logging systems that were previously impractical due to lack of processing power, practical size and cost are now available to the practitioner. An overview of the requirements of present day ambulatory data logging is presented and analogue tape, solid-state memory and disk drive storage recording systems that have been described in the literature are investigated in detail. It is proposed that digital based technology offers the best solution to the problems encountered during human based data logging. The appearance of novel digital storage media will continue the trend of increased recording durations, signal resolution and number of parameters thus allowing the momentum gained throughout the last several decades to continue. PMID- 11411254 TI - Parasitic infections among the aborigines in Taiwan with special emphasis on Taeniasis asiatica. AB - According to the results obtained by our laboratory and the Department of Parasitology, Taipei Medical College, from 1995 to 1998, the overall infection rate of enterobiasis was 16% (4-29%) among 4,426 primary school students at 64 schools in 13 mountainous districts of 10 counties, and the infection rate of head louse infestation was 12% (3-26%) among 5,121 students. A continuous series of field studies on epidemiology and chemotherapy of taeniasis asiatica have been conducted between 1971 and 1992 among 27,359 aborigines in 88 villages of 14 districts in 10 counties and 3,104 were found to be infected, giving an overall infection rate of 11%. Laboratory studies have been performed on the morphology, intermediate hosts, experimental infection, immunology and molecular biology. Moreover, development of Taiwan Taenia in one of 3 volunteers has been successfully demonstrated since 1988. Moreover, the domestic pig was identified as the intermediate host of Taiwan Taenia as in the classical T. saginata (Poland strain). Most of cysticerci were developed in or on the parenchyma of liver. The results indicated that Taiwan Taenia is different from classical T. saginata. It has been named as T. s. asiatica. Moreover, the classical T. saginata was renamed as T. s. saginata. This paper summarizes findings and reviews those reported by our laboratory. In addition, the clinical manifestations and eating habits of the aborigines are also discussed. PMID- 11411255 TI - Transcriptional mRNA of bone morphogenetic proteins 2, 3, 4, and 5 in trigeminal nerve, benign and malignant peripheral nerve sheath tumors. AB - Bone morphogenetic proteins (BMPs) have been shown to play an important role in cell growth and differentiation. BMPs, a rapidly expanding family closely related to transforming growth factor-beta (TGF-beta) superfamily, have been proven recently to possess a regulatory role and neurotrophic capacity in neurogenesis. The aim of the present study is to reveal the relationship among BMPs, peripheral nerve and neoplastic lesions of nerve sheath tumors. The mRNA transcriptions of BMP 2, 3, 4 and 5 in 12 cases of schwannoma, four cases of malignant schwannoma and three cases of trigeminal neuralgia were detected using an in situ hybridization technique. Our results demonstrated that the myelin sheaths of schwann cell from the peripheral neuroectomy of trigeminal neuralgia were positively expressing mRNA of BMP-2, 3, 4 and 5. However, the nerve fibers of trigeminal nerve showed only BMP-2 positive staining. All of the neoplastic lesions of nerve sheath showed a consistent but variant expression of BMP-2, 3, 4, and 5. Except for the BMP-4 mRNA, the expression signals of BMP-2, 3 and 5 mRNA in malignant schwannoma were relatively lower than in benign lesions. On the basis of the findings, we concluded that selected members of BMPs existed in the peripheral nerves and might contribute to the health maintenance, proliferation, regeneration and neoplastic transformation of the peripheral nerve system. Moreover, the effects of BMP-2, 3, 4 and 5 on peripheral nerve system and its neoplastic transformation might be widespread, diverse and antagonistic. PMID- 11411256 TI - Frequency doubling technology perimetry in retinal diseases--preliminary report. AB - The role of frequency doubling technology perimetry (FDT) in glaucoma has been promising in speed and simplicity as well as in its high sensitivity and specificity compared to conventional automatic perimetry. In this study, we evaluate the role of frequency doubling technology perimetry (FDT) in a variety of retinal diseases. FDT using full-threshold C-20 mode was performed in thirty three patients with a variety of retinal diseases [16 cases of rhegmatogenous retinal detachment (RRD), nine cases of age-related maculopathy (AMD) and eight cases of central serous chorioretinopathy (CSCR)]. The fundus was scored by the extent of disease on each quadrant and macular area (central five degree visual angle). Then the retinal scores were compared with the points depressed to 2% on total deviation plots in FDT C-20 full-threshold mode using Spearman's rank correlation method. The correlation was statistically significant in the group of RRD (R = 0.57, P = 0.01), whereas the correlation was not significant in the group of macular diseases (AMD & CSCR, R = 0.47 & 0.32, P = 0.12 & 0.20). In conclusion, our results suggest that FDT might be a useful and simple technique for functional evaluation in RRD, but not in the macular disease group under this scoring system. PMID- 11411257 TI - Correlation of cervical vertebra maturation with hand-wrist maturation in children. AB - The purpose of this study was to evaluate the reliability of cervical vertebra maturation as an indicator of skeletal age during the circumpubertal period. This was determined by correlating cervical vertebra maturation to hand-wrist maturation. The vertebral skeletal age was assessed using lateral cephalometric radiographs according to maturity indicators modified from Lamparski. The hand wrist skeletal age was evaluated in radiographs with the system developed by Fishman. The sample consisted of 503 subjects (244 boys and 259 girls), aged 8 through 18 years. The Spearman rank correlation coefficients and Wilcoxon sign rank test showed that a statistically significant relationship existed between the two assessments. Both the intra- and inter-judge tests of reliability displayed no significant differences. The results of this study indicate that skeletal age assessment made from the maturational changes of cervical vertebrae were reliable, reproducible and valid. PMID- 11411258 TI - Stress and maternal-fetal attachment of pregnant women during their third trimester. AB - This study was conducted to investigate the maternal-fetal attachment, pregnancy stress and life-event stress of women during their third trimester and to explore the predictors of the maternal-fetal attachment. Purposive sampling was used to recruit 150 subjects from two hospitals in Kaohsiung. Four instruments were used to collect data: (a) the Demographic Data Questionnaire, (b) the Modified Maternal Fetal Attachment Scale, (c) The Pregnancy Stress Rating Scale, and (d) the Assessment Chart for Stressful Events of Adult Life. Factor analysis of the Modified Maternal Fetal Attachment scale defined four factors: (1) interacting with the fetus, (2) giving of self, (3) identifying the fetus, and (4) fantasizing. Results of the stepwise multiple regression indicated that the best subsets to predict the criterion variable of maternal-fetal attachment, including parity, attendance of prenatal class, stress from seeking safe passage for herself and her child through pregnancy, and the life-event stress, accounted for 32% of variance. PMID- 11411259 TI - Aneurysms of the fenestrated basilar artery treated with Gulielmi Detachabe Coils: case report. AB - Multiple aneurysms at the site of a basilar artery fenestration (BAF) are extremely rare. We report a case of BAF in association with two aneurysms successfully treated with Guglielmi Detachable Coils (GDCs). A 42-year-old male presented with subarachnoid hemorrhage and consciousness change. Angiograms of the vertebrobasilar artery demonstrated fenestration of the proximal basilar artery associated with two aneurysms. Embolization of aneurysms was done by using GDCs. Aneurysms were almost completely obliterated with preservation of the flow and lumen of the vertebrobasilar system. The patient remains asymptomatic at clinical follow-up of 18 months. PMID- 11411260 TI - Inverted follicular keratosis. AB - Inverted follicular keratosis (IFK) is a benign skin lesion that typically presents as an asymptomatic, solitary nodule on the face of middle-aged and older individuals. IFK may mimic malignant lesions, especially squamous cell carcinoma (SCC), both clinically and pathologically. We report a 35-year-old male patient who had an exophytic nodule with papillary surface on his right nasal ala of 3 months' duration. The nodule was solitary, flesh colored, 5 mm in diameter and tended to bleed. The patient received shave excision with cautery under the clinical impression of pyogenic granuloma. Pathologically, diagnostic confusion existed between IFK and SCC. Squamous eddy formation and lack of epithelial dysplasia were suggestive of IFK. The tissue was investigated for the presence of human papillomavirus (HPV) DNA sequences by polymerase chain reaction (PCR) and negative result was obtained. This patient healed satisfactorily after removal of the lesion and no visible recurrence occurred during the following 10 months. The possibility of histologically confusing IFK with SCC, leading to unwarranted disfiguring treatments of a benign lesion, is emphasized. PMID- 11411261 TI - Intraoperative stroke under epidural anesthesia for bipolar hemiarthroplasty in a patient with multiple myeloma: a case report. AB - A 56-yr-old, 58-kg woman with right femoral neck fracture underwent right bipolar hemiarthroplasty. Her medical problems included diabetes mellitus and multiple myeloma. Epidural anesthesia was performed uneventfully. No other drug except for local anesthetic (2% lidocaine) was administered during the procedure. Unfortunately, we "witnessed" the dramatically hemodynamic change of an intraoperative stroke, which was preceded by a hypotensive episode and followed by sustained hypertension. The stroke was presented with decreased level of consciousness initially and confirmed by the brain CT, which revealed acute infarction at the right middle cerebral artery territory, four days after the procedure. Multiple mechanisms such as intraoperative hypotensive episode and vascular atherosclerotic change due to poor controlled diabetes mellitus might be involved in the event. In the meanwhile, hyperviscosity (3.7 centipoise units of the patient v.s. 1.4-1.8 centipoise units of normal range) caused by multiple myeloma might also have contributed to the intraoperative stroke. Clinical presentation of this case will be discussed. PMID- 11411262 TI - Micronutrient deficiency conditions in the Middle East region: an overview. AB - The Middle East is perhaps the world's best laboratory for learning more about iron and zinc deficiencies, and their widespread prevalence contributes much to constraining the quality of life for a large share of the population. Always, in the context of endemic IDD, countries in the region need to make sure that the programs are in place to consign this still-serious problem to the history books. Research is not required so much in relation to IDD, but clearly applications need to be monitored. Vitamin A deficiency is probably widespread at mild to moderate levels, and should be addressed by a combination of appropriate case management for infectious disease, breastfeeding promotion, and dietary diversification/nutrition education. Iron deficiency is the most prevalent, and so far the most intractable, micronutrient problem. It probably will not yield to less than a multipronged strategy including fortification of the food supply with iron. Several other micronutrient deficiencies require our eventual attention as well, and looking forward to their solution should be part of the research agenda. PMID- 11411263 TI - Comments on the management of iron deficiency and iron deficiency anemia. PMID- 11411264 TI - Monitoring micronutrient deficiency conditions. AB - Data for monitoring of nutritional status need to be selected and interpreted with as much care as goes into the quality control for their collection. Cost and efficiency need to be considered as well as interpretability. Few indices of nutritional status are completely specific, and those that are reasonably specific tend to be not very sensitive. As progress toward the control of micronutrient deficiency conditions occurs, the focus needs to be on early indicators of deficiency and risk, and on risk factors including dietary intake. PMID- 11411265 TI - Relationship between plasma homocysteine and vitamin status in the Framingham study population. Impact of folic acid fortification. AB - Recent studies have demonstrated associations between occlusive vascular disease and hyperhomocysteinemia of both genetic and nutritional origin. In the present study we analyzed plasma samples from the 20th biannual examination of the Framingham Heart Study cohort to determine distribution of plasma homocysteine concentrations with emphasis on relationships to vitamins that serve as coenzymes in homocysteine metabolism and to prevalence of carotid artery stenosis. Results showed that homocysteine exhibited strong inverse association with plasma folate and weaker associations with plasma vitamin B12 and pyridoxal-5'-phosphate. We saw similar inverse associations between homocysteine and intakes of folate and vitamin B6, but not vitamin B12. Prevalence of high homocysteine (> 14 mumol/L) was 29.3% in this cohort, and was greatest among subjects with low folate status. Inadequate plasma concentrations of one or more B vitamins appear to contribute to 67% of the cases of high homocysteine. Prevalence of stenosis > or = 25% was 43% in men and 34% in women, with an odds ratio of 2.0 for individuals in the highest homocysteine quartile (> or = 14.4 mumol/L) compared with those in the lowest quartile (< or = 9.1 mumol/L), after adjustment for sex, age, HDL cholesterol, systolic blood pressure, and cigarette smoking (Ptrend < 0.001). Plasma concentrations of folate and pyridoxal-5'-phosphate and folate intake were inversely associated with extracranial carotid stenosis after adjustment for age, sex, and other risk factors. Studies using samples from the Framingham Study Offspring Cohort have shown that the US-mandated folic acid fortification of flour and cereal grain products resulted in an increase in the mean folate concentrations from 4.8 to 10.0 ng/mL (P < 0.001) and prevalence of low folate (< 3 ng/mL) decreased from 22.0 to 1.7% (P < 0.001) between the baseline and follow up visits. Mean homocysteine concentration decreased from 10.1 to 9.4 microM (P < 0.001), and prevalence of high homocysteine (> 13 mumol/L) decreased from 18.7 to 9.8% (P < 0.001) between study visits. There were no statistically significant changes in the control group for folate or homocysteine between examinations. These data indicate a high prevalence of hyperhomocysteinemia in the Framingham Study population, the majority of which can be attributed to vitamin status and that this hyperhomocysteinemia is clinically relevant because of its association with increased risk of occlusive extracranial carotid stenosis. Insufficient levels of folate, and to a lesser extent vitamin B6, appear to predict part of this elevated risk through their role in homocysteine metabolism. These studies also indicate that the recently-implemented fortification of grain and cereal products with folic acid resulted in a substantial decline in plasma homocysteine. The impact of fortification on the US population is likely to be similar; however it awaits the next survey for further confirmation. PMID- 11411266 TI - Prevention of micronutrient deficiency conditions: the Israeli approach. PMID- 11411267 TI - U.S. experience with food fortification. PMID- 11411268 TI - CDC perspectives on food fortification. PMID- 11411269 TI - Health economic analysis of micronutrient supplementation: the World Bank perspective. PMID- 11411270 TI - Micronutrients for the elderly. PMID- 11411271 TI - Canadian experience with food fortification. PMID- 11411272 TI - Crusaders, cars, combinations, and cuisine. PMID- 11411273 TI - Health and nutrition communication. PMID- 11411274 TI - The first Israeli national health and nutrition survey (MABAT) as a policy maker. PMID- 11411275 TI - First Palestinian national health & nutrition survey. PMID- 11411276 TI - Negev nutritional studies: nutritional deficiencies in young and elderly populations. AB - The importance of nutrition to public health and preventive medicine is evident. Undernutrition is a main nutritional risk factor in the elderly and has been established as a cause of excess morbidity and mortality in different segments of the older population. In the infant population, inadequate nutrition is one of the causes of iron-deficiency anemia, which is associated with impaired physical and cognitive development and lowered immunity. The aim of this paper was to estimate the nutritional pattern and micronutrient deficiencies in elderly and young populations in the Negev. In southern Israel, 351 subjects over 64 years old reported mean dietary intake that was lower than that in younger persons and was independent of the presence of chronic diseases. Current data from southern Israel on healthy Jewish children revealed anemia prevalence of 15% in the second year of life. Data from recent prospective study on Bedouin children showed that anemia affected one quarter of children at age one year. Thus, infants in this area are at high risk for iron deficiency. The findings require the attention of public health authorities and food manufacturers, and should result in a range of activities including publicity and educational programs, fortification of foods, and supplementation programs in high risk-groups. PMID- 11411277 TI - Nutrient deficiency conditions in Palestine. PMID- 11411278 TI - Osteoporosis and vitamin D deficiency in Israel. PMID- 11411279 TI - Prevalence of goiter among primary school students, aged 8-10 years, in the West Bank and Gaza, 1997. PMID- 11411280 TI - Nutrition and the development of human capital. PMID- 11411281 TI - Hemoglobin levels among Arab and Druze children aged 1-2 years in Akko subdistrict. PMID- 11411282 TI - Iron deficiency effects on brain function. PMID- 11411283 TI - Anemia in infancy. PMID- 11411284 TI - [A fundamental symmetry between morphogenesis and function of branched organs]. AB - It is generally difficult to find any relationship between the morphogenesis of an organ and its final function. A priori, such a relationship has no reason to exist, since organs do not actually function during their formation. I will show in this article that, for a very large class of organs--the branched organs- there exists a hidden relationship between their morphogenesis and their function. This class of organs comprises: the lungs, the salivary mammary and lacrymal glands, the kidneys, the pancreas, and possibly other organs, such as testes. For all these organs, a fundamental fact that comes from recent developments in physics explains at the same time how they form, and why they work. This suggests, first, that complex organs are not the result of gradual and long selection processes, and, second, that this specific structure for the organs is imposed by the laws of physics. The growth process, as described here, is possibly the only one that allows both to build a fluid-secreting organ, and make it work. PMID- 11411285 TI - [Pro- and anti-apoptotic role of nitric oxide, NO]. AB - NO displays both pro- and anti-apoptotic properties. The parameters governing these effects begin to be elucidated. Among these figure the nature of the cells, their redox state, the flow and concentration of NO, its possibility to react with superoxide generated at the level of mitochondria. The targets of NO include molecules involved in DNA repair, such as PARP, the DNA-dependent protein kinase (DNA-PK) and p53 which control the transcription of various genes involved in the apoptotic process (bax, cdk inhibitors), and the proteasome which control the degradation of several apoptotic proteins. The inhibition by NO of caspases through S-nitrosylation of their active sites provides a rationale for our understanding of the anti-apoptotic effect of NO, but other mechanisms are involved, such as a regulation of the mitochondrial permeability. A better knowledge of the various steps of the apoptotic process that are affected by NO would allow the design of new pharmacological tools. PMID- 11411286 TI - 'Vertition' of integumental organs in mites revisited: a case of fluctuating asymmetry. AB - Vertition in mites is defined as a meristic variation for a bilateral integumental organ with a separate genetic control for each body side. A prominent hypothesis expressed by Grandjean is the role of vertition in the evolutionary trend towards a reduced number of hair-like organs (mechano- and/or chemo-receptors) known to have occurred in many mite groups. Observations on leg setae in the two-spotted spider mite Tetranychus urticae do not support this hypothesis. Meristic variation for leg setae rather conforms to the notion of fluctuating asymmetry: the difference between the number of leg setae on the right and left sides of the body had a unimodal distribution with a mean of zero. Moreover, lack of heritability for left/right absences of leg setae in an inbred laboratory strain suggests that vertition could be purely environmental. It is therefore argued that meristic variation for hair-like organs in mites is caused by random developmental accidents not corrected by homeostatic mechanisms normally resulting in a perfect bilateral symmetry. PMID- 11411287 TI - [Ultrastructure of Weddellomyces epicallopisma (Dothideales, Dacampiaceae, Ascomycota), especially of its ascospores]. AB - An ultrastructural study of Weddellomyces epicallopisma (ascomata wall, asci, ascospores and vegetative hyphae), the first done on the family Dacampiaceae, confirms most of the observations made in light microscopy. Moreover it shows that ascospores are provided with an endospore (not visible in light microscope) and that the structure of the ascospore septum is more complex. The similarity of the wall structure between the ascospore and the hyphoid appendages, developed on the upper part of the ascoma, is emphasized. PMID- 11411288 TI - Chorology of New Caledonian palms and possible evidence of Pleistocene rain forest refugia. AB - The distribution of the 36 palm species of New Caledonia is studied in relation to several parameters: elevation, rainfall, geological substrate, phytogeographical sectors and vegetation types. The climate (thermal gradient and rainfall) appears to be the principal factor influencing the distribution of palms, the substrate effect being subordinate to the climatic pattern. Nearly all palm populations are included within the 1,500 mm isoline. Maximum levels of species richness and endemism are located in four areas receiving more than 3,000 mm of annual rainfall. We interpret these areas as former Pleistocene refugia of lowland rain forest based on three lines of evidence: 1) all locally endemic lowland palm species and genera are restricted to these areas; 2) local endemics occur on east-facing slopes receiving the highest rainfall and most likely to have sustained rain forests during the driest periods; and 3) several pairs of sister species are disjunct between the southeastern and northeastern high rainfall areas. PMID- 11411289 TI - [Properties of the structures produced by ecology engineers at the soil surface of a Colombian savanna]. AB - Fourteen types of physical structures produced by ecosystem engineers were sampled at the surface of a savanna soil of Colombia. Invertebrates implicated in the creation of these structures were identified. Some physical (aggregate size and stability, bulk density) and chemical (C, N, P contents, pH, etc) properties of structures were assessed. Three large groups of structures were identified: (i) earthworm casts characterised by a high bulk density (1.3-1.4 g.cm-3), constituted of aggregates (7-10 mm), high in organic C (3-4%) and assimilable nutrients; (ii) termite mounds with low bulk density (0.6-0.9 g.cm-3), constituted of aggregates (8-9 mm), high in organic C (3.5-10%) and assimilable nutrients; and (iii) slightly compact (0.4-0.7 g.cm-3) and granular (aggregate size < 1.5 mm) termite superficial channels and ant mounds low in organic C (less than 1.5%) and assimilable nutrients. These results underline the large diversity of the biogenic structures produced at the surface of the studied soil. They suggest the feasibility of a functional classification of engineer organisms that would take into account simultaneously the different functional attributes reflected by these structures. PMID- 11411290 TI - [Dietary behavior of the Middle Ages in Grenoble: application of isotopic biogeochemistry of the Saint-Laurent cemetery (XIIIth-XVth centuries, Isere, France)]. AB - Isotopic analysis of 13C and 15N of 47 bones from Saint-Laurent de Grenoble cemetery (Grenoble, Isere), from the end of medieval period (XIIIth-XVth centuries AD) allowed to define the food status of animals with regard to the humans and to discuss the variability amongst adults. Adults who died young and those with small stature may have had diets poor in animal protein. The consumption of animal proteins was more important in the XVth than in the XIVth century and could illustrate a typical urban food economy providing its population with a more diversified diet than in rural areas. PMID- 11411291 TI - A putative insect intracellular endosymbiont stem clade, within the Enterobacteriaceae, infered from phylogenetic analysis based on a heterogeneous model of DNA evolution. AB - Insect intracellular symbiotic bacteria (intracellular endosymbionts, or endocytobionts) were positioned within the gamma 3-Proteobacteria using a non homogeneous model of DNA evolution, allowing for rate variability among sites, for GC content heterogeneity among sequences, and applied to a maximum likelihood framework. Most of them were found to be closely related within the Enterobacteriaceae family, located between Proteus and Yersinia. These results suggest that such a bacterial group might possess several traits allowing for insect infection and the stable establishment of symbiotic relationships and that this could represent a stem clade for numerous insect endocytobionts. Based on the estimations of the equilibrium GC content and branch lengths in the phylogenetic tree, we have made comparisons of the relative ages of these different symbioses. PMID- 11411292 TI - The oldest stegocephalian from the Iberian peninsula: evidence that temnospondyls were euryhaline. AB - The previous fossil record of limbed vertebrates of the Iberian peninsula started in the Triassic (245 Ma). The discovery of a new temnospondyl from the Late Carboniferous (Stephanian C, 290 Ma) extends the fossil record of stegocephalians in this region by at least 45 Ma. Early stegocephalians are usually thought to have been unable to live in salt water, but the new temnospondyl described below appears to have lived in a coastal region, presumably in salt water. PMID- 11411293 TI - Lung salvage and protection ventilatory techniques. AB - Physicians are in the beginning of an era in intensive care medicine in which they finally are starting to see improved outcomes in patients with AHRF. At the same time, intensivists are presented with a bewildering choice of ventilator options and adjunctive therapies. Trying to sort out which are "cosmetic," that is, improve the blood gases as opposed to influencing the outcome, remains a challenge and will be resolved only with additional RCTs. Principles of ventilator management that are driven by mimicking normal physiology are inappropriate and must be rethought. PMID- 11411294 TI - Noninvasive monitoring in the pediatric intensive care unit. AB - The best ICU monitors are physicians and nurses, who integrate all of the physiologic parameters of patients with the known pathophysiology of the disease process. Over-reliance on raw electronic data, with their inherent errors, jeopardizes the safe and efficient care of patients. Data must be interpreted in the context of the history, repetitive physical examinations, response to therapy, and a background of experience. New modalities and the application of artificial intelligence may facilitate the interpretation of data, but the role of the bedside medical practitioner remains as the heart of pediatric critical care. PMID- 11411295 TI - Point-of-care testing. AB - Point-of-care testing technology rapidly is changing the way physicians practice medicine by facilitating the availability of biochemical parameters immediately or almost immediately. The constant evolution and developments in [figure: see text] microchemistry and computer technology will make this area a dynamic part of medicine with the constant emergence of improved and newer technologies. Clinicians must not forget, however, that the best analyzer and monitor is the physician, nurse, or other health care worker in direct contact with the patient, constantly reassessing, re-examining, and integrating all of the physiologic and biochemical data in the context of the history and physical examination. If POC testing is implemented, its goal should be to improve and assist in patient care. PMID- 11411296 TI - Septic shock. AB - Infection is problematic because it affects many patients (adults and children), is a major cause of death in intensive care units (ICU) worldwide, and uses a large amount of hospital resources. The mortality rate among patients with septic shock varies but approximates 40% in infected patients admitted to ICUs. Because of the large number of adults dying of sepsis, many resources are expended. Children are physiologically different from adults, but nonetheless, many similarities exist with respect to the response to septic shock. PMID- 11411297 TI - Drowning. Rescue, resuscitation, and reanimation. AB - Several myths about drowning have developed over the years. This article has attempted to dispel some of these myths, as follows: 1. Drowning victims are unable to call or wave for help. 2. "Dry drownings" probably do not exist; if there is no water in the lungs at autopsy, the victim probably was not alive when he or she entered the water. 3. Do not use furosemide to treat the pulmonary edema of drowning; victims may need volume. 4. Seawater drowning does not cause hypovolemia, and freshwater drowning does not cause hypervolemia, hemolysis, or hyperkalemia. 5. Drowning victims swallow much more water than they inhale, resulting in a high risk for vomiting spontaneously or on resuscitation. No discussion of drowning would be complete without mentioning the importance of prevention. Proper pool fencing and water safety training at a young age are instrumental in reducing the risk for drowning. Not leaving an infant or young child unattended in or near water can prevent many of these deaths, especially bathtub drownings. Also crucial is the use of personal flotation devices whenever boating. Proper training in water safety is crucial for participation in water recreation and sporting activities, including SCUBA diving. The incidence of pediatric drowning deaths in the United States has decreased steadily over the past decade, perhaps as a result of increased awareness and attention to drowning prevention measures (Box 1). PMID- 11411298 TI - Predicting outcome in hypoxic-ischemic brain injury. AB - Predicting the neurologic outcome of children after a hypoxic-ischemic event continues to be a challenge for intensivists and pediatric neurologists. Nevertheless, with accurate history taking, serial neurologic examination, and some ancillary studies, the clinician can predict accurately whether a child will die or have profound neurologic damage. Aggressive resuscitation should be offered to all children when found in CPA. A simple ingestion might have led to this clinical scenario, and complete neurologic recovery may be possible if effective resuscitation is implemented. In cases of drowning, several factors, if present, are consistent with profound neurologic sequelae or death. These include prolonged submersions with asystole, delayed onset of CPR, no spontaneous respirations on arrival to the emergency department, and low initial pH value. The options of withdrawal of life support or a DNR status should be offered to families of children who have survived a devastating hypoxic-ischemic event but who are in a PVS. If brain-death criteria have been fulfilled, the patient must then be disconnected from life support after organ donation has been discussed with the family. PMID- 11411299 TI - Cerebral resuscitation after traumatic brain injury and cardiopulmonary arrest in infants and children in the new millennium. AB - As outlined in Figure 1, it is likely that a series of interventions beginning in the field and continuing through the emergency department, ICU, rehabilitation center, and possibly beyond, will be needed to optimize clinical outcome after severe TBI or asphyxial CA in infants and children. Despite the many differences between these two important pediatric insults, it is likely that many of the therapies targeting neuronal death, in either condition, will need to be administered early after the insult, possibly at the injury scene. Even cerebral swelling, a pathophysiologic derangement routinely treated in the PICU, almost certainly is better prevented rather than treated. Finally, this review includes, for one of the first times, a brief discussion of additional horizons in the management of patients with severe brain injury, namely, manipulation of the injured circuitry and stimulation of regeneration. Further research is needed to define better the pathobiology of these two important conditions at the bedside, to understand the optimal application of contemporary therapies, and to develop and apply novel therapies. The tools necessary to carry out these studies are materializing, although the obstacles are great. This difficult but important challenge awaits further investigation by clinician-scientists in pediatric neurointensive care. PMID- 11411300 TI - Status epilepticus. AB - Status epilepticus is a serious medical emergency that requires prompt and appropriate intervention. Maintenance of adequate vital function with attention to airway, breathing, and circulation; prevention of systemic complications; and rapid termination of seizures must be coupled with investigating and treating any underlying cause. In most patients with SE, the use of adequate dosages of first line antiepileptic agents allows for the successful and rapid termination of SE and avoidance of potential neurologic complications. Refractory SE requires more aggressive treatment, often the use of intravenous anesthetic agents and intense monitoring, and therefore must be managed in a pediatric intensive care unit with a multidisciplinary approach. Large, controlled, multicenter, comparative studies are needed urgently to clarify better the optimal management of these patients. PMID- 11411301 TI - Sedation-analgesia in the pediatric intensive care unit. AB - The provision of sedation and analgesia is an integral aspect of the care of PICU patients. A careful systems approach to the provision of sedation and analgesia can minimize complications and maximize benefit to patients. Vigilance in monitoring and adherence to published guidelines are important for safety. Physicians must define the goals in clearly devising a plan and tailor the prescription to those goals rather than use a regimented protocol for all patients. PMID- 11411302 TI - Care of the potential pediatric organ donor. AB - Although all of this information may create the impression that caring for a potential organ donor is an exceedingly complex task, in the authors' experience, this often is not true, and much energy can--and should--be devoted to the care of the bereaved family. Of crucial importance are the early recognition of brain death and the consequent radical switch of the treatment goal from preservation of the patient's brain and life to preservation of organs for the lives of others. Care for the donor is the natural extension of care for a critically ill or injured patient. During the foregoing discussion, the authors had to stress the absence of sound evidence on many points. Because many reports originate from transplant centers dedicated to a specific organ, gaining a comprehensive view on management options in the ICU further is hampered. Thus, this situation leaves another field in which investigations originating from pediatric intensivists could provide evidence urgently needed to make optimal choices. The next decade should see the thyroid hormone controversy solved by at least one controlled prospective study and the differential applicability of inotropic, vasoactive, or fluid-centered strategies. It seems self-evident that only graft survival and related parameters can form adequate endpoints for future studies. PMID- 11411303 TI - The technology-dependent child. AB - Improvements in the provision of oxygen, mechanical ventilation, tracheostomy care, enteral and parenteral nutrition, and dialysis have expanded the population of technology-dependent children. This article attempts to review pertinent points regarding these services, including common complications. Primary care and subspecialty physicians must smooth the transition of these children to the home environment, but a comprehensive team approach is necessary for the recognition of medical complications and provision of appropriate family teaching and psychosocial supports. PMID- 11411304 TI - Pharmacogenetics and pharmacogenomics. AB - This article introduces pharmacogenetics and pharmacogenomics in the context of pharmacotherapy in the pediatric ICU setting. As an independent discipline (if it can be considered as such), pediatric or developmental pharmacogenetics is essentially at a neonatal stage. Available pharmacokinetic data derived from studies of drugs that are largely dependent on a single CYP pathway for their elimination provide initial assessments of the developmental profile of that particular CYP isoform. Essentially then, pharmacogenetics in a pediatric context refers to the changes in phenotype that occur as a child grows and develops. Furthermore, the apparent drug biotransformation "phenotype" may be influenced by disease (infection), environmental factors (diet and environmental contaminants) and concurrent medications; however, drug response is a function of the complex interplay among genes involved in drug transport, drug biotransformation, receptors, and signal transduction processes, among others. Therefore, optimization of pediatric pharmacotherapy necessarily requires that developmental changes in each of these areas and not just drug biotransformation be investigated thoroughly before the promise of pharmacogenetics and pharmacogenomics for rational therapeutics can be realized in children. PMID- 11411305 TI - Coding and billing in the pediatric intensive care unit. AB - Physicians need to be proficient in their use of CPT codes and ICD-9 codes. They must participate actively and be knowledgeable of their billing process. An organized approach to coding and billing has been suggested by the American Academy of Pediatrics, as outlined in the following box. Physicians are ultimately responsible for any bill submitted in their name. Therefore, close scrutiny of the coding and billing procedures is paramount. Even if mistakes are made in the billing process beyond physicians' input, physicians still may be held responsible. If physicians do their own coding, the bills become more accurate, which can result in higher reimbursement. Physicians also should have a functional compliance plan in place, whether practicing in a large faculty group practice or practicing solo, with the ability to audit the coding and billing process and respond to variances if they are found. By being more involved in the process, physicians can have a more efficient billing system that avoids the potential for fraud and abuse and improves collections. PMID- 11411306 TI - The virtual pediatric intensive care unit. Practice in the new millennium. AB - Patients and their families meet with health care providers in a complex marketplace. The information revolution is providing access to vast amounts of information and new ways to understand it. More important, perhaps, is that it also is providing new ways of communicating information not only about health but also about the health care delivery process. This occurrence makes it possible for patients not only to diagnosis and treat themselves but also see how well the professionals do it. Like all marketplaces, asymmetries in information define the value of the interaction. Patients see physicians because they have no way of overcoming this knowledge barrier, and health care is a highly regulated market because of these asymmetries in information. New information technologies in general and telemedicine (which, in this broad sense, include distance learning for patients) can address and erode these information asymmetries. This technology threatens to have a profound effect on health care. Telemedicine offers to increase greatly the reach (connectivity) and richness (bandwidth, customization, and interactivity) of the health care information marketplace. This radically will change the way in which physicians practice critical care. Intensivists must ensure that patients continue to receive high-quality critical care. This practice will require embracing these new technologies. Resisting them will be catastrophic. What is the VPICU? It is a committed group of pediatric intensivits who are dedicated to supporting pediatric critical care medicine in the enhancement of knowledge about pediatric critical care. It includes application of information technologies to support the practice of pediatric critical care. It primarily is focused on understanding the health care delivery process and providing the tools for pediatric intensive care practitioners to better understand the care they deliver. It is the desire of the VPICU to create a virtual community in which pediatric critical care practitioners work together to understand the way they practice and to identify and implement better ways to deliver pediatric critical care. This virtual community will be responsible for clinical and economic performance in the practice of pediatric critical care. The VPICU realizes that this requires the tools to make high-quality decisions and that these decisions depend on data and communication. The author invites all pediatric intensivists to participate in the VPICU to achieve the goals of better practice through the application of information technologies in pediatric critical care. PMID- 11411307 TI - Haploid spermatids exhibit translationally repressed mRNAs. AB - During spermiogenesis, DNA-binding histones are replaced by protamines. DNA protamine-interactions result in chromatin condensation causing cessation of transcription in elongating spermatids. Thus, in haploid spermatids, processes of transcription and translation need to be temporally uncoupled in order to ensure protein synthesis in transcriptionally silent germ cells. Post-transcriptional events, such as processing, transport, and storage of mRNAs, therefore, play important roles in determining when transcripts become functionally available for translation. This review focuses on mechanisms of translational regulation in haploid spermatids and male infertility caused by defects of translational control processes. PMID- 11411308 TI - Creation of chimeric mutant axolotls: a model to study early embryonic heart development in Mexican axolotls. AB - The Mexican axolotl (Ambystoma mexicanum) provides an excellent model for studying heart development since it carries a cardiac lethal mutation in gene c that results in failure of contraction of mutant embryonic myocardium. In cardiac mutant axolotls (c/c) the hearts do not beat, apparently because of an absence of organized myofibrils. To date, there has been no way to analyze the genotypes of embryos from heterozygous spawnings (+/c x +/c) until stage 35 when the normal (+/c or +/+) embryos first begin to have beating hearts; mutant (c/c) embryos fail to develop normal heartbeats. In the present study, we created chimeric axolotls by using microsurgical techniques. The general approach was to transect tailbud embryos and join the anterior and posterior halves of two different individuals. The chimeric axolotl is composed of a normal head and heart region (+/+), permitting survival and a mutant body containing mutant gonads (c/c) that permits the production of c/c mutant offspring: 100% c/c offspring were obtained by mating c/c chimeras (c/c x c/c). The mutant phenotypes were confirmed by the absence of beating hearts and death at stage 41 in 100% of the embryos. Examination of the mutant hearts with electron microscopy and comfocal microscopy after immunofluorescent staining for tropomyosin showed identical images to those described previously in naturally-occurring c/c mutant axolotls (i.e., lacking organized sarcomeric myofibrils). These "c/c chimeric" axolotls provide a useful and unique way to investigate early embryonic heart development in cardiac mutant Mexican axolotls. PMID- 11411309 TI - The growth arrest specific gene (gas6) protein is expressed in abnormal embryos sired by male golden hamsters with accessory sex glands removed. AB - Expression of growth arrest specific gene (gas6) and its receptors in embryonic and uterine tissues in normal pregnancy and pregnancy that produces abnormal embryos sired by hamsters with partial or total deletion of male accessory sex glands was studied by in situ hybridization, immunohistochemistry, reverse transcription polymerization reaction and enzyme-linked immunoabsorbant assay. At oestrus, very strong gas6 mRNA and Gas6 expression were seen only in the uterine epithelium and endometrial glands. Upon implantation, both of them could be demonstrated in the decidualizing stroma. From day 4 to day 7 p.c, gas6 mRNA was present in the embryo, but Gas6 immunoreactivity was only found in those showing features of degeneration. The gas6:beta-actin mRNA ratio was low in oestrus and at day 4 of pregnancy but rose as the embryo grew. As for the receptors, Rse was detected in embryonic cells during days 5-7 p.c., and decidual cell from days 4 to 7 p.c., but Mer could be found in decidual cells and trophoblasts. It was concluded that gas6 had a role in endometrial transformation during decidualization and trophoblastic invasion. In the embryo, gas6 was transcribed, but the protein was only produced in response to need, such as when normal progression of development was threatened. PMID- 11411310 TI - Topography and distribution of sympathetic nerve fibers in the rat temporomandibular joint: immunocytochemistry and ultrastructure. AB - The distribution and fine structure of nerve fibers containing neuropeptide Y (NPY), tyrosine hydroxylase (TH), and vasoactive intestinal polypeptide (VIP) in the temporomandibular joint were investigated by both the avidin-biotin complex method and an indirect immunofluorescence technique. The innervation pattern of NPY- and TH-positive fibers differed from that of VIP-positive fibers. Specifically, the former was distributed in both the superficial and deep sublining layers, while the latter was mostly located in the deep sublining layer. NPY- and TH-immunoreactive fibers were largely confined to vascular elements; occasional fibers were observed in the synovial lining layer close to the joint cavity. More nerves with NPY and TH immunoreactivity were observed close to the upper joint compartment than near the lower compartment NPY and TH immunoreactivity was dramatically reduced in the TMJ of superior cervical ganglionectomized animals, indicating the sympathetic origin of these nerves. NPY immunoreactivity was found only in unmyelinated axons, which were located in the adventitia and adventitia-medial border of arteries or arterioles. Occasionally, axons were near the joint cavity, in areas free of vascular structures. These observations show that abundant sympathetic nerves supply the temporomandibular joint of the rat and provide a morphological basis for the involvement of different neuropeptides in vascular regulatory and modulatory functions in physiological and pathophysiological conditions. PMID- 11411311 TI - Expression and localization of connexin 43 in rat incisor odontoblasts. AB - We have examined the expression and localization of connexin 43 (CX43) in rat incisor odontoblasts using reverse transcriptase polymerase chain reaction, in situ hybridization and immunohistochemistry. The CX43 gene was expressed in odontoblasts, and levels of gene expression increased throughout the course of development. In contrast, CX43 was down-regulated at an incisal segment. In situ hybridization analysis showed no positive signal for CX43 RNA in the cytoplasm of differentiating dental papilla cells, but faint positive signals for CX43 RNA were observed in early pre-odontoblasts. Those signals were more intense in young and in old odontoblasts, but were less in short odontoblasts. CX43 could not be detected in differentiating dental papilla cells or in early pre-odontoblasts by immunohistochemical localization, but a positive reaction was found in the late pre-odontoblast stage where predentin had been produced. The positivity gradually increased during odontoblast maturation, and was highest in the layer of old odontoblasts. These results indicate that odontoblasts that secrete actively dentin matrix components are tightly in contact with each other by gap junctions as suggested by the intense CX43. PMID- 11411312 TI - Increase in calcitonin gene related peptide (CGRP) and decrease in mast cells in dihydroepiandrosterone (DHEA)-induced polycystic rat ovaries. AB - The polycystic ovary is reported to correspond with a high density in intraovarian nerve fibers and their sympathetic hyperresponsiveness. Peptidergic nerves may also be involved in this process. An interaction between nerve fibers and mast cells is assumed because of nerve growth-factor production by mast cells. Here we investigated CGRP-positive nerve fibers and mast cells in polycystic ovaries induced in immature rats with dihydroepiandrosterone (DHEA). The DHEA treated ovaries contained less corpora lutea than controls (mean +/- SEM: 4.3 +/- 0.6 versus 11.3 +/- 0.9, P > 0.001) and less intact antral follicles (4.7 +/- 0.7 versus 8.1 +/- 1.1; P < 0.05) according to the histometric approach. By immunolabelling more CGRP-positive nerve fibers were found in the DHEA treated ovaries than in controls (mean +/- SEM per one section: 23.2 +/- 5.8 fibers versus 10.3 +/- 0.9 and 171 +/- 44.7 varicosities versus 84 +/- 9.5). This was confirmed by dot blot analysis, showing a significant higher CGRP signal intensity per microgram homogenized ovaries of the DHEA treated group compared to the untreated (P < 0.05). Toluidine-blue-stained mast cells populated the medulla in both groups, yet had strikingly decreased in the DHEA treated ovaries (23.5 +/ 3.9 versus 89 +/- 5.6, P < 0.005). CONCLUSION: The increase in CGRP-positive nerve fibers and the decrease of toluidine-blue-stained mast cells points to an altered neuroimmune function in DHEA-induced polycystic rat ovaries. PMID- 11411313 TI - Ultrastructural localization of the NADPH-diaphorase activity in the Leydig cells of aging mice. AB - Recently, it has been shown that nitric oxide may inhibit the Leydig cell steroidogenesis. The present paper describes, by means of NADPH-diaphorase histochemistry, the ultrastructural localization of the enzyme nitric oxide synthase in the Leydig cells of young adult and aging mice. In the young adult mice, the enzymatic reaction was mainly located in the mitochondria and in some clustered cisternae of the smooth endoplasmic reticulum. The nuclear envelope was faintly labeled. In the aging mice, most Leydig cells showed an enhanced enzymatic reaction. Labeled mitochondria were increased in number, and labeled areas of the smooth endoplasmic reticulum were more numerous and extended. In addition, a strong enzymatic reaction was recognized in the nuclear envelope. We conjecture that the impaired steroidogenesis observed in the testis of aging mammals might, at least in part, depend on the increased nitric oxide production in the Leydig cells. PMID- 11411314 TI - Magnetic resonance imaging and three-dimensional reconstructions of the brain of a fetal common dolphin, Delphinus delphis. AB - To demonstrate the kinds of data that can be obtained non-destructively and non invasively from preserved museum specimens using modern imaging technology the head region of a whole body fetal specimen of the common dolphin, Delphinus delphis, aged 8-9 months post-conception, was scanned using Magnetic Resonance Imaging (MRI). Series of scans were obtained in coronal, sagittal and horizontal planes. A digital three-dimensional reconstruction of the whole brain was prepared from the coronal series of scans. Sectional areas and three-dimensional volumes were obtained of the cerebral hemispheres and of the brainstem-plus cerebellum. Neuroanatomical features identified in the scans include the major sulci of the cerebral hemispheres, well-differentiated regions of gray and white matter, the mesencephalic, pontine, and cervical flexures, the "foreshortened" appearance of the forebrain, and the large auditory inferior colliculi. These findings show that numerous features of the fetal common dolphin brain can be visualized and analyzed from MRI scans. PMID- 11411315 TI - Cortical connections of the claustrum and subjacent cell groups in the hedgehog tenrec. AB - Cyto- and chemoarchitectural findings have recently suggested that in the hedgehog tenrec, the claustrum is not located below but between the layers of the rhinal/insular cortex (Kunzle and Radtke-Schuller 2000b). The present connectional study confirms this unusual position. Tracer injections were made into various isocortical and allocortical regions. They showed that the tenrec's dorsal claustrum was reciprocally and bilaterally connected with the neocortex. The ventral claustrum was connected with mainly the ipsilateral paleocortex, additionally with the ventromedial frontal cortex and possibly the subiculum. A sparsely labeled cell group separated the claustrum from the labeled cells located in the depth of the RCx and the adjacent paleo- and neocortices. On the basis of the linear arrangement of these latter cells immediately adjacent to the subcortical white matter, and the restriction of their labeling to the ipsilateral side, one might interpret preliminarily these cells as layer 6B cells or persisting subplate neurons. Their cortical projections showed a similar topographic organization as the claustro-cortical projections. The unusual features described in tenrec were discussed with respect to similar organizations in other mammals with poorly differentiated brains and compared with embryonic brains of mammals with more differentiated brains. PMID- 11411316 TI - [Legal malpractice problem of guidelines]. PMID- 11411317 TI - [Myocardial ischemia in hypertensive patients--pathogenesis, prevalence and prognosis]. AB - Arterial hypertension causes myocardial hypertrophy and affects coronary circulation through structural and functional changes of the coronaries. Therefore, the presence of arterial hypertension predisposes to atherosclerotic coronary artery disease. We examined the frequency of myocardial ischemia as well as their changing by means of medical blood pressure reduction in the case of 104 hypertensives with insufficiently adjusted blood pressure results. The patients had a check-up before and after the suspension of normotensive blood pressure by means of a 24-hour long-term ECG. Significant results for a myocardial ischemia were horizontal and descending ST-segment depression > or = 0.1 mV for the duration > or = 1 min. Before starting the treatment there were 38 patients with 335 ischemic episodes. The percentage of silent ischemic episodes was very high (94%). After lowering blood pressure 181.0 +/- 8.8 mmHg./98.7 +/- 9.7 mmHg vs. 136.4 +/- 11.5 mmHg./78.2 +/- 14.1 mmHg a reduction of the number of ischemic episodes (median 5 vs. 0, p < 0.0001) could be shown as well as a reduction in the ST-total area per patient (median 3.8 mV.min vs. 0 mV.min, p < 0.0001)--the decisive parameter for evaluation of the grade of the severity of ischemia. The danger of ischemic attacks respectively the total ischemic burden is reduced by antihypertensive drug treatment. This is important for the prognosis. PMID- 11411318 TI - [Interdisciplinary management of chronic tinnitus (II)]. AB - Tinnitus is defined as the perception of sound in the absence of any appropriate external stimulation. It is a common, bothersome and hard-to-evaluate symptom and in most cases it cannot be objectified. Its incidence in Germany and the western world is about 10%. About 1-2% of the population are severely disturbed by tinnitus and it may disrupt everyday activities and sleep. Recent theoretical developments favour a neurophysiological approach as an explanation for tinnitus in addition to a psychoacoustic model based on peripheral lesion in the cochlea or auditory nerve. In the neurophysiological model, the processing of the tinnitus signal plays a dominant role in its detection, perception and evaluation. Therefore, attention and other psychological factors become important in the understanding and treatment of tinnitus. Many treatments of chronic tinnitus have been proposed and implemented. Today, cognitive-behavioural treatment is regarded as an important part of an integrative therapy which may be compiled of counselling, relaxation therapy, instrumental (hearing aid, tinnitus masker, tinnitus instrument, tinnitus noiser) and pharmacological tools (lidocaine, neurotransmitters). In well-controlled studies the empirical support for other therapeutical approaches such as acupuncture is weak. This work gives a review of the current knowledge of the etiology, pathogenesis, epidemiology, the interdisciplinary diagnostic approach and treatment of tinnitus and especially focuses on insurance and medico-legal aspects. PMID- 11411319 TI - [Bank robbery as trauma--"but who understands me?"]. AB - The different ways victims behave and feel after having experienced a bank robbery as well as some of the usual questions they may ask will be described using a typical case. The symptoms produced by a trauma do not only require a treatment based on empathy but also special attention. Should you ever have to deal with a traumatized victim of a robbery, the first and most important thing is not to leave the person alone. Simply talking to the person, without any reproaches may already help to calm him/her down. The most helpful thing is to involve the family, relatives and friends in further care. If the victim does not react when talking to him/her and counselling does not produce relief, then medication is advisable. If the persons have been taken hostage, or in the case of strong symptoms, it is advisable to seek the assistance of a psychologist or a psychiatrist. Therefore, dealing with victims after a bank robbery can be compared with a balancing act on a tightrope. A lot of sensitivity and patience are required. PMID- 11411320 TI - [Mobbing--significance for private personal insurance]. AB - The social conflict model called mobbing has hitherto almost remained undetected by private personal insurance systems. The mental and functional somatic disturbances caused by mobbing, however, have been well known for a long time and do not differ from those caused by other conflicts. Problems for the personal insurance develop from the fact that more often than not a (feigned) conflict solution will be sought by medical certifications of unfitness to work, referrals to hospitals and striving for acknowledgement of occupational disablement, none of which have a medical justification. A considerable portion of the national economic burden of mobbing will therefore--mostly unknowingly--be shouldered by private insurance companies. There are numerous medical diagnoses which may hide a mobbing conflict and are difficult to comprehend. This article represents instructions for better understanding and handling. PMID- 11411321 TI - [Patient role in changing conditions: yesterday--today--tomorrow]. AB - Changing conditions within the German healthcare system and new trends of information management will decisively change the patient's role. Increase in morbidity caused by the ageing population aggravates the economic pressure within the social healthcare system. Due to the ongoing medical progress, this will inevitably lead to the restriction of medical services trough budgeting. Whereas the patient's role in the past was rather passive, his position will change in future. This is not only caused by an increase in patient education and social status, but also by the availability of information provided by the worldwide web. The number of educated patients who are gathering information via the worldwide web and discussing their diseases with fellow patients via e-mail will rapidly grow. The traditional doctor-patient relationship will be less exclusive in future and will be embedded in the network of reimbursement facilities, patients' groups and information management. PMID- 11411322 TI - [Hospital hygiene I: Resolution by AWMF and hospital hygiene II: Experts campaign for health protection]. PMID- 11411323 TI - Doctor proactive communication, return-to-work recommendation, and duration of disability after a workers' compensation low back injury. AB - Although doctors are increasingly evaluated on the basis of return-to-work (RTW) outcomes, the effect of doctor-patient communication about the workplace and RTW after an occupational injury has received little research attention. The effect of patient-reported doctor communication on duration of disability was examined retrospectively in a 3-year cohort of 325 claimants with a lost-time low back injury. Although doctor proactive communication was associated with a greater likelihood of RTW during the acute phase (< 30 days of disability), this effect disappeared when injury and workload characteristics were taken into account. A positive RTW recommendation was associated with about a 60% higher RTW rate during the subacute/chronic phase (> 30 days of disability) only. Prospective studies are needed to confirm this effect. The impact of physician communication on RTW is largely confounded by injury and workplace factors. PMID- 11411324 TI - Applying genomic technologies in environmental health research: challenges and opportunities. AB - Recent discoveries in molecular biology and genetics have made it possible for environmental health researchers to examine how genetic characteristics affect response to environmental exposures. Understanding such gene-environment interactions offers exciting possibilities for the prevention and control of environmentally induced diseases. Despite these potential benefits, the collection and analysis of genetic information in environmental health research presents many of the same ethical, legal, and social (ELSI) challenges found in other types of genetic research. In this article, we describe a number of ELSI challenges in environmental genomic research and the opportunities and responsibilities that accompany this research. PMID- 11411325 TI - The association of health risks with workers' compensation costs. AB - The purpose of this study was to investigate the association between health risks and workers' compensation (WC) costs. The 4-year study used Health Risk Appraisal data and focused on 1996-to-1999 WC costs among Xerox Corporation's long-term employees. High WC costs were related to individual health risks, especially Health Age Index (a measure of controllable risks), smoking, poor physical health, physical inactivity, and life dissatisfaction. WC costs increased with increasing health risk status (low-risk to medium-risk to high-risk). Low-risk employees had the lowest costs. In this population, 85% of WC costs could be attributed to excess risks (medium- or high-risk) or non-participation. Among those with claims, a savings of $1238 per person per year was associated with Health Risk Appraisal participation. Addressing WC costs by focusing on employee health status provides an important additional strategy for health promotion programs. PMID- 11411326 TI - Risk factors for work-related injury among male farmers in Iowa: a prospective cohort study. AB - Agriculture is one of the most hazardous occupations in the United States. We examined the work-related injury rate and risk factors among 290 Iowa male principal farm operators using a population-based, prospective study. Baseline data were collected between 1991 and 1994, and injury events were surveyed in the fall of 1995. Cumulative incidence of farmwork-related injury during the previous year was 10.5%. We found that depressive symptoms (odds ratio, 3.22; 95% confidence interval, 1.04 to 9.99) and the number of hours working with animals (odds ratio, 2.14; 95% confidence interval, 1.04 to 4.44) were associated with the incidence of farmwork-related injuries. PMID- 11411327 TI - Occupational sunlight exposure and mortality from non-Hodgkin lymphoma among electric utility workers. AB - This case-control study examined occupational sunlight exposure and death from non-Hodgkin lymphoma (NHL) and NHL subtypes among 188 cases and 1880 controls selected from a cohort of 138,905 male electric utility workers. Exposure was classified according to work history linked to indices of cumulative sunlight exposure. Odds ratios and 95% confidence intervals were derived from conditional logistic regression models and were conditioned on the matching factors birth year and ethnicity. Mortality from NHL and intermediate/high-grade lymphomas was not related to cumulative sunlight exposure, with odds ratios around the null. For low-grade lymphomas, a dose-response gradient was observed for exposure in the past 12 to 21 years, but this result seemed to be sensitive to cut points for categorization of cumulative exposure. These data do not provide evidence for an association between occupational sunlight exposure and mortality from NHL or NHL subtypes. PMID- 11411328 TI - Preventing needlesticks in emergency medical system workers. AB - Emergency medical system (EMS) workers frequently use sharp devices in injury prone circumstances that involve limited visibility, confined spaces, rapidly moving vehicles, and uncooperative victims. This study examined the efficacy of an automatic self-retracting lancet in reducing needlestick injuries and related direct and indirect costs. Subjects were 477 active-duty EMS workers. Counseling, laboratory testing (hepatitis B and C, hepatic function enzymes, and human immunodeficiency virus), antiviral prophylaxis, and immunizations were provided according to US Public Health Service guidelines. Baseline and biennial laboratory testing for hepatitis B and C and liver function enzymes were conducted. After the introduction of a spring-loaded automatic-retracting type glucometer lancet device, needlestick injuries decreased from 16 per 954 EMS worker-years to 2 per 477 EMS worker-years. The annualized cost of treatment declined from $8276 to $2068. The change to a self-retracting device decreased the number of needlestick injuries and was cost-effective with a minimal increase in device cost (annualized $366 per year). PMID- 11411329 TI - Risk factors for adverse health effects following hazardous materials incidents. AB - To identify risk factors for persistent morbidity, we conducted a prospective study of individuals involved in hazardous materials incidents reported to the Washington Poison Center. Between December 1997 and October 1999, 202 subjects in 87 incidents were surveyed by telephone. Medical symptoms persisting for a minimum of 8 days after the incident were reported in 51 (25%) subjects, and 18 (9%) left work or school for more than 2 days because of the exposure. Medical intervention was reported in 46 (58%) of 79 subjects for whom medical records were available, and objective abnormalities were found in 57 (72%). Multiple logistic regression analysis indicated that subjects with dermal exposures, three or more alcoholic drinks per week, and previous use of psychiatric medications were more likely to report persistent symptoms. Divorced, widowed, or separated subjects, asthmatic subjects, and those having initial dermal symptoms were more likely to miss work or school for more than 2 days. Of patients evaluated at a health care facility, subjects with preexisting hypertension were more likely to receive medical treatment or have objective medical findings, whereas those with inhalation exposures and those decontaminated at the scene were less likely to be treated or have abnormalities. In our study, both incident and individual factors were predictive of adverse health effects, and these findings should be considered in planning the care of patients involved in hazardous materials incidents. PMID- 11411330 TI - Effects of exposure to organic solvents on menstrual cycle length. AB - To investigate the association between organic solvent exposure and menstrual disturbance, we conducted a cross-sectional study among 1408 petrochemical workers in China. Based on an industrial hygiene evaluation, we classified the workshops according to the presence or absence of organic solvents (benzene, styrene, toluene, or xylene). We used logistic regression to estimate odds ratios and 95% confidence intervals for prolonged menstrual cycle length (oligomenorrhea: average cycle length > 35 days during the previous year) associated with the exposure. After adjustment for confounders, each additional year of work in an exposed workshop was associated with a 7% increase in oligomenorrhea (odds ratio, 1.07; 95% confidence interval, 1.00 to 1.14). Compared with no exposure, 3 or more years of exposure was associated with a 53% increase in oligomenorrhea (odds ratio, 1.53; 95% confidence interval, 1.00 to 2.34). We concluded that exposure to organic solvents is associated with a trend toward increased frequency of oligomenorrhea. PMID- 11411332 TI - Editorial concerning the examination process. PMID- 11411331 TI - Respiratory findings in livestock farmworkers. AB - In this cross-sectional study, the prevalence of acute and chronic respiratory symptoms and lung function were investigated in 236 (169 male and 67 female) livestock farmworkers raising domestic animals, including cattle and horses. The prevalence of acute and chronic respiratory symptoms was recorded by administering a modified version of the British Medical Research Council questionnaire on respiratory symptoms. Lung function was measured by recording the maximum expiratory flow-volume curves from which the forced vital capacity, the one-second forced expiratory volume, and the maximum expiratory flow rates at 50% and the last 25% were measured. There was a significantly higher prevalence of almost all chronic respiratory symptoms in farmers compared with controls: highest among men for chronic cough (27.2%) and highest among women for dyspnea (28.3%). Typical symptoms of occupational asthma were diagnosed in 3.6% of the male and 1.5% of the female livestock farmers. A logistic regression analysis performed on chronic respiratory symptoms of livestock workers indicate significant effects of smoking (P < 0.001) on all symptoms except occupational asthma. A large number of male and female livestock farmworkers complained of acute work-related symptoms: highest for dry cough (52.2%) and shortness of breath (44.9%). Lung function data among these workers demonstrated a trend toward lower lung function in general. Multivariate analysis of lung function parameters indicate significant effects of length of employment and smoking in male and length of employment in female livestock farmworkers. The data suggest that employment in livestock farming may be associated in a dose-dependent manner with the development of acute and chronic respiratory symptoms and lung function changes. PMID- 11411335 TI - It's the law. PMID- 11411334 TI - Profiling the abusive personality. PMID- 11411336 TI - Reporting child abuse--it's not a matter of choice. PMID- 11411337 TI - Lasers in dentistry. PMID- 11411338 TI - Dental photography. PMID- 11411339 TI - Dental informatics: a new career in dentistry. PMID- 11411340 TI - Digital radiography: state of the art. PMID- 11411341 TI - [Physiological and pharmacological function of PPARs]. AB - The finding of nuclear receptors has greatly enhanced our understanding of gene regulation by lipophilic hormones such as steroids, thyroxine, vitamin D and retinoids. These receptors comprise a superfamily of transcription factors containing highly related DNA-binding domains. In mammals, the peroxisome proliferator-activated receptor (PPAR) family of nuclear hormone receptors consists of three subtypes by separate genes: PPAR alpha, PPAR delta (also referred to as hNUC1 or PPAR beta), and PPAR gamma. PPARs have been associated with several distinct biological programs. PPARs function as a heterodimer with the retinoid X receptor. This complex binds to sequences termed direct repeat-1 response element in enhancer sites of regulated genes and activates transcription upon ligand and coactivator binding. Three different PPAR subtypes have specific roles in different organs. PPAR alpha, mainly expressed in liver, plays an important role in fatty acid metabolism. PPAR gamma predominantly is expressed in adipose cells. PPAR delta displays a high level of expression in lipid metabolizing organs such as small intestine, heart and adipose tissue. Naturally occurring and synthetic molecules (anti-hyperlipidemia and diabetic drugs) that are ligands for these nuclear receptors control transcriptional activity of PPARs. We believe that the pharmacological and genomic researches on PPAR will develop powerful tools for prevention and medical care against common diseases. PMID- 11411342 TI - [Role of central histamine in amygdaloid kindled seizures]. AB - The role of central histamine in amygdaloid kindled seizures in rats was studied. Histamine content in the amygdala was significantly decreased after development of amygdaloid kindling. Intracerebroventricular (i.c.v.) injection of histamine resulted in inhibition of amygdaloid kindled seizures. The H1-agonists 2 methylhistamine and 2-thiazolylethylamine also inhibited amygdaloid kindled seizures. In addition, intraperitoneal (i.p.) injection of histidine and metoprine inhibited amygdaloid kindled seizures at doses that caused increases in histamine contents of the brain. H1-antagonists (diphenhydramine and chlorpheniramine) attenuated histamine- or histidine-induced inhibition of amygdaloid kindled seizures. Both i.c.v. and i.p. injections of H3-antagonists (thioperamide, AQ0145 and clobenpropit) resulted in a dose-related inhibition of amygdaloid kindled seizures. The effects of thioperamide and AQ0145 were inhibited by an H3-agonist (R)-alpha-methylhistamine and H1-antagonists. On the other hand, H2-antagonists showed no antagonistic effect. GABAmimetic drugs, diazepam, sodium valproate and muscimol potentiated the effect of clobenpropit. Bicuculline caused significant antagonism of the inhibition of amygdaloid kindled seizures induced by clobenpropit. These findings suggested that a histaminergic mechanism plays an important role in suppressing amygdaloid kindled seizures through histamine H1-receptors. In addition, an inhibition of amygdaloid kindled seizures induced by histamine is closely related with the action of GABA. PMID- 11411343 TI - [Insulin resistance-reducing effect of a new thiazolidinedione derivative, pioglitazone]. AB - Takeda has a relatively long history in diabetes research. Pioglitazone, a thiazolidinedione derivative, was developed from our basic research on diabetic animal models in the 1960s and our chemical research on lipid-lowering agents in the 1970s. Pioglitazone reduced plasma glucose, triglyceride and insulin levels in obese-diabetic animal models with insulin resistance in liver and/or peripheral tissues, but did not decrease normoglycemia in normal rats and aged dogs or hyperglycemia in insulin-deficient streptozocin-induced diabetic rats and impaired-insulin-secretory Goto-Kakizaki rats. The ED50 of plasma glucose lowering action was 0.5 mg/kg/day in Wistar fatty rats. These findings clearly indicate that pioglitazone works in animals with insulin resistance and has a quite different mechanism from sulfonylureas and insulin itself. Although the exact mechanism of pioglitazone still remains obscure, pioglitazone normalized abnormalities in the cellular signal transduction of insulin. These effects seem to be due to the inhibitory action of pioglitazone on TNF-alpha production, which is one of the factors responsible for insulin resistance. Pioglitazone is a potent agonist for the peroxisome proliferator-activated receptor, (PPAR)-gamma, that is related to differentiation of adipocytes, and the relationship between TNF-alpha production and PPAR-gamma has been reported. Therefore, the agonistic activity of pioglitazone on PPAR-gamma may be involved in the mechanism of reduction of insulin resistance. The clinical data clearly demonstrated that pioglitazone, at clinical doses of 15-45 mg/day, decreased plasma glucose, HbA1c and triglyceride, increased plasma HDL-cholesterol, but did not alter total cholesterol and LDL-cholesterol levels. These findings suggest that pioglitazone has a benefit for prevention of cardiovascular diseases in addition to diabetic complications. PMID- 11411344 TI - [Pharmacological and pharmacokinetic properties of azithromycin (Zithromac), a novel 15-membered ring macrolide antibacterial agent]. AB - Azithromycin (Zithromac), a 15-membered ring macrolide antibacterial agent, was approved to be manufactured in Japan in March 2000. It showed good in vitro and/or in vivo antibacterial activities against Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Peptostreptococcus micros, Haemophilus influenzae, Moraxella catarrhalis, Mycoplasma pneumoniae and Chlamydia pneumoniae. Its activity against H. influenzae was particularly more potent than that of currently used macrolide antibacterial agents. After oral administration to patients, azithromycin was readily absorbed and became widely distributed throughout the body, achieving higher concentrations in tissues and phagocytic cells than in serum or plasma. Its distribution into phagocytes was as high as more than 10 times that of erythromycin, and azithromycin was readily released from phagocytes in the presence of S. aureus. In experimentally infected mice, the concentration of azithromycin was higher in infected tissues than in uninfected tissues, which indicated that azithromycin was selectively delivered to infected tissues by migrating phagocytes. These pharmacological and pharmacokinetic properties were reflected in good clinical results for the treatment of respiratory infections and other infections with once daily dosing for 3 days. PMID- 11411345 TI - [Pharmacological profile and clinical effect of tacrolimus ointment (Protopic ointment) for atopic dermatitis]. AB - Atopic dermatitis (AD) is thought to be induced by a complex of various allergic reactions and T cells are implicated in its etiology. Since tacrolimus strongly inhibits T cell activation, tacrolimus ointment has been developed as a novel drug for AD throughout the world. Tacrolimus inhibits mast cell and eosinophil activation and antigen presenting activity of Langerhans cells in vitro. In the in vivo experimental animal models of AD, such as contact and spontaneous dermatitis in mice and repeated hapten treated skin inflammation in rats, tacrolimus ointment showed inhibitory activity. In clinical studies with AD patients in Japan, USA and Europe, tacrolimus ointment showed a marked effect. In comparative studies in Japan, it showed the same efficacy as a strong class steroid ointment on eczema at the trunk and extremities and superior efficacy at the face and neck compared to a medium class steroid. The most prominent adverse event is experienced at the local application site with reactions such as a burning sensation and erythema. Systemic side effects were rarely observed. While there is a possibility of skin infections when using tacrolimus, skin atrophy, even after long term treatment, was not observed. Thus tacrolimus ointment could be an efficient alternative to steroid ointment for AD. PMID- 11411346 TI - [The development of new drugs for osteoporosis in Japan]. AB - In a rapidly aging society, the number of patients becoming bed-ridden due to osteoporosis-related fracture has become a socially important health issue that includes the problem of adequate nursing. Recently, the introduction of a rapidly acting bone absorption suppressant with a clear mechanism accelerated the development of osteoporosis treatment. A selective estrogen receptor modulator in a estrogen preparation is expected, because it has antagonism for the acceptor of the reproductive organ. It is noted that a new steroid with a weak androgen action and estrogen- and progestogen-like actions both prevents bone quantity decrease in women and increases bone quantity in osteoporotic patients after menopause. The second and third generation of bisphosphanate is more powerful than etidronate, the first generation of bisphosphanate, allows continual medication, and is now in clinical trial. The introduction of new drugs, which have clear efficacy and fewer side effects may be expected, along with the combined use of the drugs in which action mechanisms differ. The introduction of pharmaceutical new approaches is also expected as the expression mechanism of osteoporosis is further clarified by basic studies. PMID- 11411347 TI - [Prospects for use of computers in the Air Force medical service]. AB - The article describes the questions for the formation of the Automated Control Sub-system of the AF Medical service (ACS AF MS) and shows basic trends towards its creation. The authors take into account specific peculiarities of the Air Force activities, as well as the modern standards of automated systems. The ACS AF MS must be integrated to the Joint Automated Control System of the Armed Forces Medical Service. The article gives the main functions of this sub-system, its information and program procedure, computer technique fitting out process for command installations. The main tasks of the whole chain of command of AF Medical service were also analysed. PMID- 11411348 TI - [The use of computers in medical examination of servicemen]. PMID- 11411349 TI - [Use of an automated workstation for clinical and immunological studies and immunodiagnosis in AIDS]. AB - A two-staged test for immune status by methods described by L. A. Kozhemyakin et al. (1987), as well as AIDS immunodiagnosis were conducted with the help of automated working place (AWP). Application of AWP will make it possible to obtain up to 150 indexes. The principle of AWP functional algorithm is based upon menu bar selection or initial data input information, and contributes to well-founded methods of clinical immunology. PMID- 11411350 TI - [Computer application in business games at the military department of medical school]. AB - Computerized practical games are considered to be an effective method that could intensity and enhance the quality of training for medical service reserve officers. At the military chair of Kursk State Medical Institute a fund of practical games was created. It includes final themes on the courses of organization and tactics of medical service, military toxicology and NBC medical defense. Practical games develop students' ability to analyse combat, logistic and medical situation, help them make a correct assignment of medical forces and means, or take opportune steps in decision-making. PMID- 11411351 TI - [Deontological problems in the earthquake epicenter]. PMID- 11411352 TI - [Syndrome of the primary response to irradiation (a literature review)]. PMID- 11411353 TI - [Changes in Regulations on medical examination in the Armed Forces of the USSR]. PMID- 11411354 TI - [Methodology of a clinical expert conclusion in military medical expertise (Part 1)]. PMID- 11411355 TI - [Treatment of pulmonary thromboembolism with small doses of streptokinase combined with heparin, nitroglycerin, and prednisolone]. AB - The efficiency of method worked out by the authors for pulmonary thromboembolism management surpasses the existing routine schemes of treatment by standard doses of streptokinase. The opportune application of this method gives positive results in considerable percentage of cases, i.e. in massive and submassive thromboembolism with acute or recidivating course. PMID- 11411356 TI - [Healing and relapse in duodenal bulb chronic ulcer in patients of young age]. PMID- 11411357 TI - [Use of domestic spirographs for examination of external breathing by the 'flow volume' loop method using computers]. PMID- 11411358 TI - [Efficacy of contrykal in acute pneumonia]. AB - The article studies quininogenase system and plasma inhibitors in patients with acute pneumonia, as well as the changes that take place under the influence of proteinase inhibitor--contrycal. For the control group of patients contrycal was not applicated. This medicine was used in combination with antibiotics and other traditional therapeutic means. Positive effect of contrycal is connected with its regulation influence upon the system of quininogenase and plasma inhibitors. Results obtained during this research give the possibility to recommend a wide application of contrycal for treatment of nondestructive forms of acute pneumonia. PMID- 11411359 TI - [Climate adaptation in patients with chronic bronchitis and its effect on efficacy of the sanatory treatment]. PMID- 11411360 TI - [New regulations for medical support in the Soviet Army and Navy]. PMID- 11411361 TI - [Predicting of the flu outbreaks in military groups]. PMID- 11411362 TI - [Medical and sociological diagnostics at various stages of military duty execution]. PMID- 11411363 TI - [Military pharmacy as an object of sanitary control]. PMID- 11411364 TI - [Characteristics of physiological response of the human body to muscular efforts on control pedals]. PMID- 11411365 TI - [Diagnosis and military medical expertise in stomach or duodenal ulcer in seamen at enlisted service]. PMID- 11411366 TI - [The outstanding Soviet surgeon]. PMID- 11411367 TI - [Remembrance of the Teacher]. PMID- 11411368 TI - [Effect of electromagnetic pulse from the nuclear explosion on electrical medical equipment]. PMID- 11411369 TI - [Current state and trends in development of automated systems in the military health care]. AB - Nowadays computer technique is used by command echelons of medical service in central structures, armed services, military regions, groups of troops, fleets, and by about 80 medical establishments of central or regional subordination. Automated systems projection and its fitting out for central structures of medical service occupy the leading place in this process. At present more than 150 complex tasks are being performed with the help of computer programs or undergo working out process. However, computer application programs sometimes are based upon ineffective medical methodics. The employment of computer facilities for carrying out primitive tasks is not unusual. There are cases of unorganized creation of programs which are not compatible in technical, mathematical or informational aspects. There are few tasks oriented to employment of personal computers. The level of researches in automatization is low. The main contents of automatization works for the nearest 4-5 years must be oriented to the creation of the basic automated information and calculation systems for data processing that could operate, before all, in the interests of medical units and establishments of central chain of command. PMID- 11411370 TI - [Responsibility of medical personnel for violation of their professional obligations (part 2)]. PMID- 11411371 TI - Cough in children. PMID- 11411372 TI - Clinical approach to a patient with cough. AB - A number of disorders of the respiratory tract and some even outside the respiratory tract can cause cough. A systematic approach towards a patient of chronic cough consisting of detailed history, physical examination of upper as well as lower respiratory tract, complete blood counts, tuberculin test, chest X ray, and peak flow rate testing will give the diagnosis in majority of children. Pulmonary tuberculosis and asthma are the two commonest conditions diagnosed. If the initial work up is inconclusive, further laboratory testing and imaging studies should be considered. Thus, radiolabelled milk scan, barium swallow and 24-hour pH monitoring would diagnose gastroesophageal reflux. Spirometry, methacholine/exercise challenge test or a therapeutic trial may be required for confirming bronchial asthma. Flexible bronchoscopy is useful for evaluation for suspected aspiration syndromes and any anatomical or dynamic problem of the airway (e.g. tracheomalacia). Spiral and high resolution computed tomography (HRCT) along with magnetic resonance imaging are the modern day imaging techniques used for studying mediastinal masses, airway obstruction and even lung parenchyma (HRCT). Sputum examination for type of cells and bacteria can be useful, especially if pseudomonas or acid-fast bacilli are identified. Pseudomonas suggests cystic fibrosis (an uncommon disease in India) which can be confirmed by sweat chloride test and gene mutation studies. PMID- 11411373 TI - Cough and asthma. AB - Cough is a common symptom in office practice. Though troublesome, it serves to maintain normal function of respiratory tract. Chronic or recurrent cough may be caused by variety of diseases, asthma being the most common amongst them. Cough, wheeze and breathlessness are classical features of asthma syndrome. Many diseases may lead to this syndrome. Asthmatic children present with cough of variable intensities and patterns. At times, wheeze and breathlessness may not be clinically apparent. It was well known that all that wheezes is not asthma but now it is well understood that every asthmatic child does not wheeze. In an acute attack of asthma, cough often starts at the end of wheezing episode. It leads to expulsion of thick, stringy mucus often in the form of casts. Though cough is a minor symptom during acute attack, it ensures removal of secretions and avoid complications. Cough is a prominent symptom in persistent asthma especially between acute exacerbations. Episodic nocturnal cough may be the only symptom of chronic asthma. Children with cough variant asthma do not wheeze. It is postulated that they have milder degree of airway hyperresponsiveness and higher wheezing threshold. However, they show all the characteristics of asthma on laboratory tests. Cough represents bronchial hyperresponsiveness and is not a measure of asthma. Hence it may be caused by many diverse etiologies such as gastroesophageal reflux, enlarged adenoids, sinusitis or tropical eosinophilia. Cough in such conditions mimicks asthma and relevant tests may be necessary for proper diagnosis. PMID- 11411374 TI - Approach to recurrent respiratory infections. AB - Rational approach to diagnosis and management of recurrent respiratory infections is needed, or else the child is subjected to unnecessary investigations and multiple drugs. Repeated respiratory symptoms do not mean a respiratory infection. A diagnosis of viral infection does not justify prescription of an antibiotic. Recurrent viral infections are part of the growing up process of any child. Giving antibiotics at every episode to cover "so-called superadded bacterial infections" will lead to "recurrent antibiotics" and adverse effects on growth. Systematic approach should be used to find the underlying cause. An otoscopic examination of a child should form part of a pediatric examination in all cases of respiratory infections. Antibiotics should be judiciously chosen depending on age, socioeconomic status, severity of infection and the type of organism expected and always given in adequate doses and proper duration. Treatment should be specific and symptomatic. Adequate drainage of the sinuses is an important adjuvant therapy. Use of cough syrups with various combinations should be avoided. Efforts should be made to diagnose and treat manifestations of hyperactive airway or allergy, role of CEA (cough equivalent asthma) and WLRI (Wheeze associated lower respiratory infections). Investigations are needed in recent lower respiratory infections and adverse effect on growth, school performance, abnormal physical findings. CBC, CRP, ESR, nasal smear, appropriate cultures, tests for TB, X-Rays, barium studies, milk scan, ultra sound, CT, MRI, bronchoscopy in selected cases. PMID- 11411375 TI - The applied physiology of cough. AB - The cough reflex is such an important defense mechanism of the respiratory tract that a thorough knowledge of its applied physiology in respiratory disease is essential for clinicians. It is a well-integrated reflex, which has afferent limb consisting of receptors and afferent nerves, the central cough center in the brainstem and the efferent limb consisting of motor nerves supplying the muscles of coughing. The cough process consists of inspiratory phase, compressive phase and expiratory phase. Stimuli that can initiate the cough process can be central or peripheral in the lungs or outside the lungs. The important function of the cough reflex is to maintain the airways and alveoli clear and healthy. The mechanisms by which it achieves this are (i) high velocity of expiratory gas flows produced in the expiratory phase (ii) the compression of the lungs and airways by high positive pleural pressure generated in the compressive phase. Cough threshold can be determined by giving acetic acid, citric acid or capsaicin inhalation challenge test. This has practical application in clinical research. The pathophysiological basis of different types of cough encountered in clinical practice is discussed. PMID- 11411376 TI - Pharmacological control of cough. AB - Cough constitutes an important medical problem in all ages. Although treatment of underlying pathologies, e.g., bronchial asthma, upper respiratory tract infection, etc. is well justified to cure cough, non-specific therapy in the form of a number of antitussive agents like, codeine, dextromethorphan, etc. is widely practiced to suppress it. Most of these agents are efficacious and well tolerated. Nevertheless, CNS toxicities may occur in new-borns, especially those with immature metabolic profile. Some recent reports about interactions (involving hepatic cytochrome P450 enzymes) between dextromethorphan and other drugs are also noteworthy. PMID- 11411377 TI - Management of community acquired respiratory infections. AB - Respiratory tract infections (RTIs) are the most common diagnosis in children visiting outpatient departments in both developing and developed countries. The impact of these infections is enormous in terms of total morbidity and mortality. Management of these infections requires a meticulous evaluation and a comprehensive but rational use of supportive, symptomatic and specific therapy. Antibiotics are widely prescribed for the treatment of community acquired respiratory infections to compensate uncertainties of diagnosis and prognosis; unnecessary anti-microbial drug therapy is expensive and resistance by respiratory pathogens is increasing at an exponential rate at a time when new anti-viral as well as anti-bacterial agents are being introduced into the therapeutic armamentarium. Major dilemmas persist regarding symptomatic therapy and the indications of antimicrobial agents. A comprehensive but brief review along with latest protocols and advancements is the management of community acquired respiratory infections in children in cited. PMID- 11411378 TI - Chronic obstructive pulmonary disease in children. AB - Chronic obstructive pulmonary disease is not a well defined entity in children. A child presenting with chronic cough and wheezing should be investigated for asthma, recurrent aspiration airway compressions, chronic infection, cystic fibrosis and immune deficiency. If a specific cause is not identified; search should be made for environmental factors such as passive smoking, air pollution and irritants. The therapeutic option for patients with chronic productive cough without specific etiology include control of environmental factors, bronchodilators and chest physiotherapy. PMID- 11411379 TI - Vaccine for India in the new millennium. PMID- 11411380 TI - Polio eradication and after: does IPV have a role? PMID- 11411381 TI - Considerations for HAV vaccine in India. PMID- 11411382 TI - Varicella vaccine--after 25 years. PMID- 11411383 TI - Haemophilus influenzae vaccine and its role in India. PMID- 11411384 TI - Pneumococcal vaccines in India. PMID- 11411385 TI - Combination vaccines: an exciting world. PMID- 11411386 TI - Immunization in special circumstances. PMID- 11411387 TI - Polio eradication in India: how far are we from reaching the goal? PMID- 11411388 TI - Desirability and feasibility of hepatitis B vaccine in EPI. PMID- 11411389 TI - Is a wellness incentive money well spent? PMID- 11411390 TI - The truth about older workers. PMID- 11411391 TI - Why young workers get hurt. PMID- 11411393 TI - LTC coverage: poised for takeoff. PMID- 11411392 TI - Asthma management in the workplace. PMID- 11411395 TI - DataWatch. The drawbacks of provider choice. PMID- 11411394 TI - Power to the people. PMID- 11411396 TI - A novel two-dimensional model for colloid transport in physically and geochemically heterogeneous porous media. AB - A two-dimensional model for colloid transport in geochemically and physically heterogeneous porous media is presented. The model considers patchwise geochemical heterogeneity, which is suitable to describe the chemical variability of many surficial aquifers with ferric oxyhydroxide-coated porous matrix, as well as spatial variability of hydraulic conductivity, which results in heterogeneous flow field. The model is comprised of a transient fluid flow equation, a transient colloid transport equation, and an equation for the dynamics of colloid deposition and release. Numerical simulations were carried out with the model to investigate the colloid transport behavior in layered and randomly heterogeneous porous media. Results demonstrate that physical and geochemical heterogeneities markedly affect the colloid transport behavior. Layered physical or geochemical heterogeneity can result in distinct preferential flow paths of colloidal particles. Furthermore, the combined effect of layered physical and geochemical heterogeneity may result in enhanced or reduced preferential flow of colloids. Random distribution of physical heterogeneity (hydraulic conductivity) results in a random flow field and an irregularly distributed colloid concentration profile in the porous medium. Contrary to random physical heterogeneity, the effect of random patchwise geochemical heterogeneity on colloid transport behavior is not significant. It is mostly the mean value of geochemical heterogeneity rather than its distribution that governs the colloid transport behavior. PMID- 11411397 TI - Influence of seaward boundary condition on contaminant transport in unconfined coastal aquifers. AB - Contaminant transport in coastal aquifers is complicated partly due to the conditions at the seaward boundary including seawater intrusion and tidal variations of sea level. Their inclusion in modelling this system will be computationally expensive. Therefore, it will be instructive to investigate the consequence of simplifying the seaward boundary condition by neglecting the seawater density and tidal variations in numerical predictions of contaminant transport in this zone. This paper presents a comparison of numerical predictions for a simplified seaward boundary condition with experimental results for a corresponding realistic one including a saltwater interface and tidal variations. Different densities for contaminants are considered. The comparison suggests that the neglect of the seawater intrusion and tidal variations does not affect noticeably the overall migration rate of the plume before it reaches the saltwater interface. However, numerical prediction shows that a more dense contaminant travels further seaward and part of the solute mass exits under the sea if the seawater density is not included. This is not consistent with the experimental result, which shows that the contaminant travels upwards to the shoreline along the saltwater interface. Neglect of seawater density, therefore, will result in an underestimation of the exit rate of solute mass around the coastline and fictitious migration paths under the seabed. For a less dense contaminant, neglect of seawater density has little effect on numerical prediction of migration paths. PMID- 11411398 TI - Transport of a decay chain in homogenous porous media: analytical solutions. AB - With the aid of integral transforms, analytical solutions for the transport of a decay chain in homogenous porous media are derived. Unidirectional steady-state flow and radial steady-state flow in single and multiple porosity media are considered. At least in Laplace domain, all solutions can be written in closed analytical formulae. Partly, the solutions can also be inverted analytically. If not, analytical calculation of the steady-state concentration distributions, evaluation of temporal moments and numerical inversion are still possible. Formulae for several simple boundary conditions are given and visualized in this paper. The derived novel solutions are widely applicable and are very useful for the validation of numerical transport codes. PMID- 11411399 TI - Limitations on the role of incorporated organic matter in reducing pesticide leaching. AB - The use of organic amendments has been suggested as a method of controlling pesticide leaching through soils. The enarenados soils of the intensive horticulture of the Almeria province of southern Spain contain buried organic matter horizons above a soil layer amended with clay. This region is ideal for understanding the potential for and limitations of organic amendments in preventing pesticide pollution. This study measured the sorption and degradation potential of carbofuran in this soil system and the hydrological behaviour of the soil horizons. The sorption of carbofuran was controlled by the organic carbon content, the degradation was strongly pH-dependent and the acidic organic layer protected the sorbed carbofuran against degradation. Hydrologically, the soil system is dominated by ponding above an amended clay layer and by the presence of macropores that can transport water through this clay. A simple model is proposed on this basis and shows that although high levels of dissolved organic carbon can be released by buried organic horizons, the major control on re-release of sorbed pesticide is the potential for sorption hysteresis in this organic layer. A comparison of sorption and degradation data for carbamate insecticides used in the region with groundwater observations for these compounds shows that no amount of incorporated organic would protect against pollution from highly water-soluble compounds. PMID- 11411400 TI - Transport of volatile compounds in porous media in the presence of a trapped gas phase. AB - The presence of an immobile gaseous phase in an otherwise-saturated porous medium affects the transport of volatile compounds. The linear theory of partitioning tracers suggests that a volatile tracer introduced into such a system should be retarded with a constant retardation factor. Using high concentrations, however, the saturation of the gaseous phase will change as an effect of the tracer test itself. Competitive gas transfer among all volatile compounds and the change of saturation may lead to tracer concentrations that are temporarily higher than those injected. We analyze the system in the framework of the coherence theory by Helfferich [Soc. Pet. Eng. J. 21 (1) (1981) 51]. The governing equations are formulated as functions of total concentration, i.e., the mass of solutes in all phases per pore volume. Neglecting dispersion and mass-transfer kinetics, we derive the characteristic form of the resulting system of hyperbolic equations. In a system with N volatile compounds, a variation of the concentrations splits up into N waves, each traveling with its own characteristic velocity. If the presence of a gaseous phase is sustained, one wave will be a standing one. We perform numerical model calculations for tracers with various Henry's law coefficients and show that the results agree with the semi-analytical solution obtained by coherence theory. PMID- 11411401 TI - Predictive modelling of NAPL injection tests in variable aperture spatially correlated fractures. AB - In preparation for a field experiment where a NAPL will be injected into a fractured sandstone aquifer, a 2D invasion percolation model of DNAPL migration in a single horizontal fracture with varying aperture has been developed. This simulation investigated the effect of spatially correlated fracture aperture fields on pressure-saturation relationships as a function of variable aperture mean, standard deviation, and spatial correlation statistics under hydrostatic conditions. Results from spatially correlated variable aperture fields can be significantly different from random fields. Longer ranges decreased entry pressures and higher standard deviations decreased nonwetting phase saturations. Mean aperture is the major control on displacement pressure, entry pressure and the form of the pressure-saturation curve. Simulation results using statistical parameters for a variable aperture natural sandstone fracture as described by Yeo et al. [International Journal of Rock Mechanics and Mining Sciences 35 (1998) 1051] closely resemble a Brooks-Corey pressure-saturation function, and exhibit a distinct entry pressure followed by a rapid increase in nonwetting phase saturation. Graphical estimates of entry pressure provide a good approximation of the critical aperture controlling the formation of a continuous nonwetting phase pathway in a variable aperture fracture. Consequently, we show that multiphase flow concepts developed for porous media can successfully be applied to variable aperture fractures. Entry pressure correlates well to the mean aperture in these simulations when using a Gaussian distribution of fracture aperture. Interpreted aperture distributions from NAPL injection experiments do not fit the true distribution well at low nonwetting phase saturations, but do at higher saturations above the entry pressure. Consequently, true, mechanical aperture variation within a fracture plane cannot be determined from NAPL injection experiments either in the field or laboratory. PMID- 11411402 TI - Chromium transport, oxidation, and adsorption in manganese-coated sand. AB - We examine how the processes of advection, dispersion, oxidation-reduction, and adsorption combine to affect the transport of chromium through columns packed with pyrolusite (beta-MnO2)-coated sand. We find that beta-MnO2 effectively oxidizes Cr(III) to Cr(VI) and that the extent of oxidation is sensitive to changes in pH, pore water velocity, and influent concentrations of Cr(III). Cr(III) oxidation rates, although initially high, decline well before the supply of beta-MnO2 is depleted, suggesting that a reaction product inhibits the conversion of Cr(III) to Cr(VI). Rate-limited reactions govern the weak adsorption of each chromium species, with Cr(III) adsorption varying directly with pH and Cr(VI) adsorption varying inversely with pH. The breakthrough data on chromium transport can be matched closely by calculations of a simple model that accounts for (1) advective-dispersive transport of Cr(III), Cr(VI), and dissolved oxygen, (2) first-order kinetics adsorption of the reduced and oxidized chromium species, and (3) nonlinear rate-limited oxidation of Cr(III) to Cr(VI). Our work supplements the limited database on the transport of redox-sensitive metals in porous media and provides a means for quantifying the coupled processes that contribute to this transport. PMID- 11411403 TI - The velocity of DNAPL fingering in water-saturated porous media: laboratory experiments and a mobile-immobile-zone model. AB - Dense nonaqueous phase liquids (DNAPLs) are immiscible with water and can give rise to highly fingered fluid distributions when infiltrating through water saturated porous media. In this paper, a conceptual mobile-immobile-zone (MIZ) model is presented to describe the structure of a DNAPL finger in water-saturated porous media and the velocity of finger propagation. A finger is composed of a finger body and a tip. The finger body has a mobile core and an immobile sheath. All the DNAPL within the tip of a finger is mobile. Lab experiments utilizing image analyses of a DNAPL (PCE) penetrating into water-saturated homogeneous glass beads were carried out in a two-dimensional transparent chamber. The results show that the fingers elongated almost linearly with time. The fingers did not grow laterally after the tip of the finger had passed. The average finger diameters were between 3.9 and 5.4 mm for PCE propagation in water-saturated glass bead porous media with mean particle diameters from 0.32 to 1.36 mm. The estimated mobile core diameters were 51-60% of the average finger diameters. PMID- 11411404 TI - Bring it on! PMID- 11411405 TI - Time to do it right: soda pop decay is a preventable disease. PMID- 11411406 TI - Soft drinks: hard on teeth. PMID- 11411407 TI - The journey back. Part Three: Back to school. PMID- 11411408 TI - Three-dimensional chess in a rowboat. PMID- 11411409 TI - Forecasting the start of the pollen season of Poaceae: evaluation of some methods based on meteorological factors. AB - The pollen of anemogamous plants is responsible for half the allergic diseases, that is to say a prevalence of 10% in the French population. Poaceae produce the first allergenic pollen almost everywhere. The work described in this article aimed to validate forecast methods for the use of physicians and allergic people who need accurate and early information on the first appearance of pollen in the air. The methods were based on meteorological parameters, mainly temperature. Four volumetric Hirst traps were used from 1995 to 1998, situated in two departments of Burgundy. Two of the methods tested proved to be of particular interest: the sum of the temperatures and the sum of Q10 values, an agrometeorological coefficient integrating temperature. A multiple regression, using maximum temperature and rainfall, was also performed but it gave slightly less accurate results. A chi 2-test was then used to compare the accuracy of the three methods. It was found that the date of onset of the pollen season could be predicted early enough to be useful in medical practice. Results were verified in 1999, and the research must be continued to obtain better statistical validity. PMID- 11411410 TI - The influence of climatic variability on the quality of wine. AB - In this paper, the quality of the wine from the Dao (Viseu) region of Portugal is examined and relationships between wine quality and climatic variability are obtained using spectral and correlation analysis to determine the structure of the temporal variations. The spectra of the series of quality of wine values show statistically significant oscillations coherent with those found in the series of teleconnection indices. The series cover a period of 33 years. A significant correlation was obtained between wine quality and minimum air temperature in May, December and total precipitation in April. The teleconnection circulation indices are used to provide some physical insight into the most significant oscillating components of the climatic and the wine quality series. We found significant and positive correlations between the quality of the wine and the Southern Oscillation Index (SOI) of August and negative with the SOI of January and with the North Atlantic Oscillation of April. Wine quality and climatic series can be predicted using statistical models depending on significant oscillations. PMID- 11411411 TI - Environmental temperatures and the incidence of food poisoning in England and Wales. AB - The incidence of food poisoning in England and Wales has been increasing for many years and it is now a major public health problem. Superimposed on this general rising trend is a well-established tendency for the number of cases of food poisoning to rise during the summer when warm weather favours the multiplication of pathogenic micro-organisms. This paper shows that weekly notifications of food poisoning in England and Wales are strongly associated with environmental temperatures, but that there are some important time lags in this relationship. The number of cases of food poisoning in a given week was only weakly correlated with the temperature of that week and the one preceding it. This suggests that factors operating close to the point of consumption within or outside the home are not the principal cause of the rise in food poisoning associated with warm summer conditions. There was a much stronger association with temperatures 2-5 weeks earlier, pointing to the importance of factors operating earlier in the food production or distribution system. The results of this study suggest that the food poisoning problem requires action by food producers and distributors as well as by consumers. PMID- 11411412 TI - Influence of weather and climate on subjective symptom intensity in atopic eczema. AB - The frequent clinical observation that the course of atopic eczema, a skin disease involving a disturbed cutaneous barrier function, is influenced by climate and weather motivated us to analyse these relationships biometrically. In the Swiss high-mountain area of Davos the intensity of itching experienced by patients with atopic eczema was evaluated and compared to 15 single meteorological variables recorded daily during an entire 7-year observation period. By means of univariate analyses and multiple regressions, itch intensity was found to be correlated with some meteorological variables. A clear-cut inverse correlation exists with air temperature (coefficient of correlation: 0.235, P < 0.001), but the effects of water vapour pressure, air pressure and hours of sunshine are less pronounced. The results show that itching in atopic eczema is significantly dependent on meteorological conditions. The data suggest that, in patients with atopic eczema, a certain range of thermo-hygric atmospheric conditions with a balance of heat and water loss on the skin surface is essential for the skin to feel comfortable. PMID- 11411413 TI - Relationship between atmospheric pressure and mortality in the Madrid Autonomous Region: a time-series study. AB - This study set out to determine the possible influence of variations in atmospheric pressure on mortality in the Madrid Autonomous Region (MAR), taking into account the possible confounding effect of other atmospheric variables. The study was based on daily mortality data from the MAR Revenue and Excise Authority, meteorological data from Getafe Observatory and air pollution data from the Madrid Municipal Automatic Air Pollution Monitoring Grid. A time-series analysis was performed, using Box-Jenkins modelling and controlling for the respective confounding variables. Furthermore, the different variables studied (pressure, temperature, pollutants, etc.) were used to produce a multivariate model of the different causes of mortality. A significant association was found between anticyclonic trend and mortality with circulatory causes in the medium long term and anticyclonic trend and mortality with respiratory causes in the long term. PMID- 11411414 TI - Fatty acid composition of brown adipose tissue in genetically heat-tolerant FOK rats. AB - The phospholipid fatty acid composition of brown adipose tissue (BAT) was examined in inbred heat-tolerant FOK rats and compared with that in conventional Wistar rats not previously exposed to heat. The FOK rats showed higher unsaturation states, as indicated by higher levels of polyunsaturated fatty acids and a higher unsaturation index and polyunsaturated fatty acids/saturated fatty acids ratio. This higher level of unsaturation was characterized by the higher amount of polyunsaturated fatty acids such as linoleic acid, arachidonic acid and docosahexaenoic acid. It may be concluded that the increased docosahexaenoic acid level in BAT phospholipids brings about the hyperplasia of BAT, causing an enhancement of its in vivo thermogenic activity as well as the systemic non shivering thermogenesis observed in heat-tolerant FOK rats. PMID- 11411415 TI - Outdoor clothing: its relationship to geography, climate, behaviour and cold related mortality in Europe. AB - It has been suggested, that the inhabitants of northern European regions, who experience little cold-related mortality, protect themselves outdoors by wearing more clothing, at the same temperature, than people living in southern regions where such mortality is high. Outdoor clothing data were collected in eight regions from 6583 people divided by sex and age group (50-59 and 65-74 years). Across Europe, the total clothing worn (as assessed by dry thermal insulation and numbers of items or layers) increased significantly with cold, wind, less physical activity and longer periods outdoors. Men wore 0.14 clo (1 clo = 0.115 m2 K W-1) more than women and the older people wore 0.05 clo more than the younger group (both P < 0.001). After allowance for these factors, regional differences in insulation and item number were correlated (r = -0.74, P = 0.037; r = -0.74, P = 0.036 respectively), but not those in clothing layers (r = -0.21; P = 0.61), with indices of cold-related mortality. Cold weather most increased the wearing of gloves, scarves and hats. The geographical variation in the wearing of these three together items more closely matched that in cold-related mortality (r = -0.89, P = 0.003). A possible explanation for this may be that they protect the head and hands, where stimulation by cold greatly increases peripheral vasoconstriction causing a rise in blood pressure that procedure haemoconcentration and raised cardiovascular risk. PMID- 11411416 TI - The role of temperature in the onset of the Olea europaea L. pollen season in southwestern Spain. AB - Temperature is one of the main factors affecting the flowering of Mediterranean trees. In the case of Olea europaea L., a low-temperature period prior to bud development is essential to interrupt dormancy. After that, and once a base temperature is reached, the plant accumulates heat until flowering starts. Different methods of obtaining the best-forecast model for the onset date of the O. europaea pollen season, using temperature as the predictive parameter, are proposed in this paper. An 18-year pollen and climatic data series (1982-1999) from Cordoba (Spain) was used to perform the study. First a multiple-regression analysis using 15-day average temperatures from the period prior to flowering time was tested. Second, three heat-summation methods were used, determining the the quantities heat units (HU): accumulated daily mean temperature after deducting a threshold, growing degree-days (GDD): proposed by Snyder [J Agric Meteorol 35:353-358 (1985)] as a measure of physiological time, and accumulated maximum temperature. In the first two, the optimum base temperature selected for heat accumulation was 12.5 degrees C. The multiple-regression equation for 1999 gives a 7-day delay from the observed date. The most accurate results were obtained with the GDD method, with a difference of only 4.7 days between predicted and observed dates. The average heat accumulation expressed as GDD was 209.9 degrees C days. The HU method also gives good results, with no significant statistical differences between predictions and observations. PMID- 11411417 TI - EBRI 2001 Retirement Survey: Retirement Confidence Retirement Survey (RCS), Minority RCS, and Small Employer Retirement Survey (SERS). PMID- 11411418 TI - The relevance of developmental-psychobiological metatheory to developmental neuropsychology. AB - This article reviews 4 aspects of developmental-psychobiological metatheory that are of particular relevance to developmental conceptions of neuropsychology: probabilistic epigenesis, the various roles of experience in affecting the development of the nervous system, and the developmental principles of equifinality and equipotentiality, the latter being especially pertinent to the understanding of compensatory phenomena observed in studies of early brain damage. PMID- 11411419 TI - A developmental-psychobiological approach to developmental neuropsychology. AB - Although both developmental psychobiology and developmental neuropsychology examine the interface between biological and psychological processes, they differ in conceptual framework. This article argues for the incorporation into developmental neuropsychology of certain aspects of the conceptual framework of developmental psychobiology. Three principles of dynamic psychobiological interaction are described and applied to four issues in neuropsychology (handedness, sex differences in behavior, critical periods, and modularity of structure-function relations). Then, it is proposed that developmental psychobiology can make four direct contributions to developmental neuropsychology. Finally, it is argued that the value of the conceptual framework provided by developmental psychobiology depends, in part, on how well it translates into procedures that can be applied in the clinical settings of the developmental neuropsychologist. PMID- 11411420 TI - The Rey figure in Amazonia: effects of jungle living on childrens' copy performance. AB - This exploratory case study investigated the organizational strategies and visual motor integration skills of 6 children living in the Amazon rainforest of Peru. As expected, a developmental trend was observed for age as older children generally required less time to draw a complex design and produced better renditions than younger children. Unexpectedly, all children, including those below the age of 8, utilized the left side of the design as an anchor in adopting a left to right strategic approach to task solution. Results are discussed in terms of potential sociocultural and neurobiological influences on brain organization, development of cognitive abilities, and potential for cultural bias when evaluating ethnic minority children. PMID- 11411421 TI - Lateralized effects on reaching by children. AB - This study examined the limb selection profiles of children for a reach-to-grasp task presented in various positions of hemispace. Underlying questions focused on the use of attentional information and lateralized effects in motor programming for reaching movements. As expected, both right- and left-handed groups used their dominant limb more frequently at the midline and in their own ipsilateral hemispace. However, in response to stimuli presented in contralateral (to the dominant limb) hemispace, both groups switched to using their nondominant limbs at significant levels. As a general comparison, right-handers exhibited greater use of their dominant limb, but arguably, motor dominance in this context may have intervened with the participant's ability to use attentional information to produce a more efficient response. Overall, these findings address the phenomenon associated with motor dominance and use of attentional information in programming. PMID- 11411422 TI - The influence of binocular visual deprivation on the development of visual spatial attention. AB - This article examines the effects of visual input on the development of attention by comparing normal children to children, all more than 8 years old, who had been treated for bilateral congenital cataracts during infancy. In Experiment 1, patients pushed a button as soon as they detected a target that appeared 100, 400, or 800 msec after a central cue. The cue either validly cued the upcoming location or invalidly cued the wrong location. Patients (n = 16) performed normally at the 100 msec and 400 msec stimulus onset asynchrony (SOA). However, when the cue preceded the target by the 800 msec SOA, patients' reaction times were not affected by the validity of the cue, especially when deprivation had extended past 4 months of age. In Experiment 2, patients indicated which of two shapes appeared in the periphery 400 msec after a central cue, with those shapes surrounded by compatible or incompatible distractors. Patients (n = 15) differed from age-matched controls in (a) being slowed more by incompatible distractors on invalid trials, and (b) tending to show a larger than normal effect of the validity of the cue preceding targets in the upper visual field. Together, these findings suggest that the normal development of attention is influenced by early visual experience. PMID- 11411423 TI - Central auditory processing of durational changes in complex speech patterns by newborns: an event-related brain potential study. AB - In this study, newborns' ability to discriminate durational changes in the fricative /s/ within a nonsense word was investigated. The results showed that infrequent increments and decrements of a speech sound duration elicit a mismatch negativity kind of response in sleeping human newborns. In the auditory event related potential to these deviant stimuli two negative waves of this response were revealed. The first negative wave peaked at about 150 msec and the second at about 350 msec after the change onset. At least one negative deflection, which was interpreted as evidence for stimulus change-detection, was observed in every infant. PMID- 11411424 TI - Event-related potential evidence for age-related differences in attentional allocation during a source monitoring task. AB - Event-related potentials (ERPs) were recorded while older and younger adults were engaged in a source monitoring task. After studying a list of words, participants were presented with a recognition test during which some of the new words were repeated, rendering them as familiar as the study words. Instructions at test indicated whether the goal was to select the previously studied words or the repeated test items. Behaviorally, the younger adults were less likely to make source monitoring errors. ERPs, averaged only for correct trials, indicated that younger adults produced late positivities of greatest amplitude in response to whichever word type was designated as target irrespective of its familiarity. The ERPs of the older adults were generally less differentiated and their late positivities greater for recently repeated words irrespective of target designation. These results suggest that source monitoring in young adults is facilitated by their ability to allocate and withdraw attention from stimuli on the basis of task relevance rather than familiarity alone, and that this attentional flexibility declines with age. PMID- 11411425 TI - Haemodynamic evaluation of tubal and male factors of infertility in natural and ovarian stimulation cycles. AB - PURPOSE: To determine the haemodynamic parameters in women with tubal and male factors of infertility and to investigate the effect of ovarian stimulation on the blood flow indices of the uterine and ovarian arteries. METHODS: Prospective evaluation of the pulsitility index (PI) and resistance index (RI) of the uterine and ovarian arteries in the tubal and male factors of infertility was done. Comparisons were made between the natural and ovarian stimulation cycles. RESULTS: In natural cycles, uterine PI was 3.55 +/- 1.39 and RI was 0.95 +/- 0.05 in tubal infertility. These were significantly lower (uterine PI = 4.13 +/- 0.08; uterine RI = 0.99 +/- 0.06) than the corresponding indices in patients with male factor infertility. After ovarian stimulation in tubal factor, uterine PI and RI were 2.62 +/- 0.73 and 0.88 +/- 0.08 and were similar (PI = 2.55 +/- 0.62, RI = 0.87 +/- 0.06) to male factor infertility. CONCLUSIONS: The data points to measurable differences in the uterine PI and RI between the tubal infertility and male infertility in the natural cycles. These differences were abolished after ovarian stimulation. PMID- 11411426 TI - Updating quality control assays in the assisted reproductive technologies laboratory with a cryopreserved hamster oocyte DNA cytogenotoxic assay. AB - PURPOSE: Despite advances in assisted reproduction, there is no progress in quality control bioassays. The objectives were to develop a comet assay to measure DNA fragmentation in thawed cryopreserved oocytes and compare this assay with one-cell mouse embryo bioassay. METHODS: Thawed hamster oocytes from a commercial source were incubated in culture media with either 0-, 50-, or 100 microM hydrogen peroxide, or, in media exposed to different contact materials and unknown proficiency analytes. Incubation time was 1.5 h at 37 degrees C. The oocytes were dried, fixed, stained with acridine orange, embedded in a mini agarose layer and electrophoresis was carried out. Fluorescent images were analyzed. The results were compared with standard one-cell mouse assay data. RESULTS: The 100-microM hydrogen peroxide treatment caused greatest DNA fragmentation in the hamster oocytes at Hours 1 and 2. A dose response was observed. Intraassay coefficient of variation was 5.7%. Only one of the five materials tested passed both assays. The data for the unknown proficiency analytes were similar for both assays. CONCLUSIONS: The oocyte comet assay demonstrated DNA fragmentation in the presence of toxic substances. The detection of toxicity in two materials that passed the mouse bioassay suggested increased sensitivity in the new assay. The oocyte comet assay and the mouse bioassay results matched in the proficiency test. However, more studies are still needed to determine optimal sensitivity. PMID- 11411427 TI - The influence of prefreeze growth rate and blastomere number on cryosurvival and subsequent implantation of human embryos. AB - PURPOSE: To determine whether the relatively low implantation rate of cryopreserved Day 2 embryos with only 2 blastomeres can be increased as a consequence of increasing their blastomere content by extending the prefreeze culture time. METHODS: Of a total of 3480 Day 2 embryos studied, 1921 (55.2%) had reached the 4-cell stage by 40 h postinsemination (FAST) and were transferred or cryopreserved. The remaining embryos that underwent subsequent cell division by 46 h (INTERMEDIATE; 18.3% of total) or 66 h (SLOW; 20.3% of total) were also cryopreserved whereas the 6.2% that remained arrested at 66 h were discarded. Thawed embryos from each category were assessed for survival, post-thaw cleavage, and implantation. RESULTS: The proportion of thawed embryos that survived, the proportion of surviving embryos that underwent post-thaw cleavage, and the implantation rate of transferred embryos were all reduced in the slower growing cryopreserved embryos. CONCLUSIONS: The growth rate, and not the number of blastomeres per se, is a critical factor in predicting the developmental potential of cryopreserved embryos. PMID- 11411428 TI - Uterine leiomyomas and their effect on in vitro fertilization outcome: a retrospective study. AB - PURPOSE: The effect of uterine leiomyomas on the outcome of in vitro fertilization (IVF) treatment has been controversial. This study was undertaken to clarify influence of fibroids on IVF success, in a large population with age and other potential confounding variables controlled for in the analysis. METHODS: A population of 141 patients with and 406 without leiomyomata undergoing their first IVF cycle was studied. RESULTS: The association between uterine leiomyomas and assisted reproduction treatment outcome was not statistically significant (OR = 0.73, 95% CI: 0.49-1.19, p = 0.21) after controlling for age and other risk factors. Also, fibroids neither affected the risk of spontaneous abortion (OR = 1.06, 95% CI: 0.44-2.60) nor the risk of ectopic pregnancy (OR = 0.78, 95% CI: 0.08-8.02). Location of fibroids (intramural vs. submucosal/subserosal) and their size had no significant effect on pregnancy outcome. CONCLUSIONS: Results from our analyses indicated that in vitro fertilization outcome was not affected by the presence of uterine leiomyomas. Therefore, in patients with normal uterine cavities and fibroids less than a certain size (i.e., < 7 cm), undergoing myomectomies as a prerequisite for assisted reproduction treatment is seriously questionable. PMID- 11411429 TI - Seminal plasma and IVF potential. Biochemical constituents of seminal plasma of males from in vitro fertilization couples. AB - PURPOSE: Many components of seminal plasma play a role in sperm motility by serving as energy sources. Human seminal plasma contains over 30 proteins, including forward motility proteins, antifertility proteins, and coagulation/liquefaction proteins. This study was designed to determine any correlation between motility or fertilization rates and concentrations of fructose, lactic acid, citric acid, carnitine, and protein in human seminal plasma. METHODS: Fertilization rates were determined by in vitro methods. Fructose, lactic acid, citric acid, and carnitine concentrations were ascertained using high performance liquid chromatography. Protein concentration was determined by Bradford assay. RESULTS: Protein concentrations were significantly different as a function of sperm motility levels. Other constituents of human seminal plasma showed an overall correlation, though not significant. No constituent exhibited significant differences as a function of fertility levels. CONCLUSIONS: Protein concentration was significantly lower for samples with high motility. No significant differences between fertility levels and constituents measured were found. PMID- 11411430 TI - Antral follicle assessment as a tool for predicting outcome in IVF--is it a better predictor than age and FSH? AB - PURPOSE: The purpose of this study is to determine if baseline antral follicle assessment may serve as additional information in predicting in vitro fertilization outcome. METHODS: Prospective, descriptive preliminary study of in vitro fertilization outcome. From July 1998 to July 1999, 224 patients underwent antral follicle assessment (follicle 2-6 mm in diameter) on baseline of the planned, stimulated in vitro fertilization cycle. The outcomes were analyzed with respect to antral follicle assessment (< or = 6 or > 6), basal cycle day 3 follicle stimulated hormone (< or = 10 or > 10 IU/L) and maternal age (< or = 35 or > 35 years). RESULTS: The clinical pregnancy rate was significantly higher in the group with baseline antral follicle > 6 compared to that in the group with antral follicle < or = 6 (51% vs. 19%, respectively). Controlling for patient age, and basal follicle stimulated hormone, the pregnancy rate was significantly higher in the group with antral follicle > 6 compared to that in the group with antral follicle < or = 6. The cancellation rate was significantly increased with advancing maternal age, elevated basal follicle stimulated hormone levels, and baseline antral follicle < or = 6. The cancellation rate was significantly higher in the group with antral follicle < or = 6 compared to that in the group with antral follicle > or = 6 (33% vs. 1%, respectively). CONCLUSIONS: In vitro fertilization outcome is strongly correlated with both maternal ages, basal cycle, day 3 follicle, stimulated hormone, and antral follicle assessment. Antral follicle assessment was a better predictor of in vitro fertilization outcome than were age or follicle stimulated hormone. Antral follicle assessment may provide a marker for ovarian age that is distinct from chronological age or hormonal markers. PMID- 11411431 TI - Births after transfer of zona-free blastocysts in oocyte donation cycles. AB - PURPOSE: Oocyte donation is a well-established method of assisted reproduction for women with irreversible infertility and with previous implantation failures after in vitro fertilization. Although the pregnancy rates are very high, sometimes implantation does not occur even after various attempts. We report the first two cases of transfer of zona-free blastocysts in oocyte donation cycles that developed to normal pregnancies and births. METHODS: The patients had undergone three previous standard oocyte donation cycles with failure of implantation. Endometrium preparation was performed after suppression of the pituitary function, with E2 valerate and Progesterone at the day of oocyte retrieval. Normally fertilized embryos were cultured in Earle's culture medium until Day 3 and in S2 medium until Day 5. For each patient, the zonae of two fully expanded blastocysts were enzymatically removed with 0.5% pronase. Zona free blastocysts were transferred for the patients 2 h later. RESULTS: On Day 12 after transfer, pregnancies were confirmed with elevated serum levels of beta hCG. A gestational sac with a foetal heart beat was seen by ultrasound 15 days later, in each patient. Normal healthy babies were born at 38 and 39 weeks of pregnancy. CONCLUSIONS: This is the first report of successful pregnancies and births after oocyte donation and transfer of zona-free blastocysts in human. It not only shows the feasibility of the treatment but also opens a new alternative for the patients with repetitive implantation failure after OD cycles. PMID- 11411432 TI - Influence of swim-up method on the recovery of spermatozoa from different types of semen samples. AB - PURPOSE: To compare the effectiveness of swim-up method, using different types of semen samples. METHODS: In this retrospective study undertaken in university medical college infertility centre, subfertile couples undergoing Intra Uterine Insemination were selected. A total of 600 semen samples used for the preparation of sperm samples using swim-up method were analyzed. Relative Yield was calculated from the sperm count and motility before and after swim up from each semen sample in six different groups. RESULT(S): Statistically significant increase in relative yield was found in oligospermic samples (20.41) followed by teratospermia (16.98). However, relative yield was low in asthenospermic (11.97) and normal (> 60 million/ml) samples (11.66). CONCLUSION(S): Semen samples with good sperm count resulted in poor recovery after swim up. Swim-up method appears to be effective for oligospermic samples. Modifications like multiple tube swim up, using appropriate incubation time based on the initial semen parameter, will enhance the sperm yield after swim up. PMID- 11411433 TI - The effects of follicular fluid and platelet-activating factor on motion characteristics of poor-quality cryopreserved human sperm. AB - PURPOSE: To evaluate the effects of follicular fluid and platelet-activating factor on sperm motion characteristics of cryopreserved oligospermic and normospermic samples. METHODS: Sperm motion characteristics were evaluated prior to cryopreservation, immediately after thawing and following incubation in human tubal fluid, follicular fluid, or 1-microM platelet-activating factor cultures. Sixteen oligospermic samples and 20 normospermic samples were examined. Sperm motion characteristics were analyzed manually according to WHO criteria (1999) and also with an automated videomicrography system. RESULT(S): Incubation in follicular fluid increased overall motility and the percentage of sperm with fast progressive motility in normospermic but not oligospermic samples. Incubation with platelet-activating factor increased overall motility and the percentage of sperm showing nonprogressive motility in both oligospermic and normospermic samples. CONCLUSION(S): The stimulatory effects of culture in follicular fluid as seen in normospermic samples do not show a significant benefit in oligospermic cryopreserved samples. Platelet-activating factor and follicular fluid increase motility via different mechanisms. Incubation of oligospermic cryopreserved sperm with PAF increases the number of motile sperm, thereby enabling easier identification of viable sperm for intracytoplasmic sperm injection in samples with severe asthenozoospermia. PMID- 11411434 TI - Sperm retrieval, fertilization, and pregnancy outcome in repeated testicular sperm aspiration. AB - PURPOSE: To report the outcome of sperm retrieval and results after ICSI in up to six repeated testicular sperm aspiration procedures. METHODS: Twenty-two men with obstructive and thirty-four men with nonobstructive azoospermia underwent 50 and 91 needle aspirations, respectively. Sufficiency of spermatozoa for ICSI and cryopreservation, fertilization rate, and pregnancy outcome was analyzed retrospectively. RESULTS: No major differences were found in sperm recovery or pregnancy outcome in the repeated cycles. Testicular aspirate containing motile spermatozoa with maintained fertilizing capacity was obtained in up to six repeated procedures in the nonobstructive group. No postoperative complications were reported for any of the participants. CONCLUSIONS: Testicular sperm aspiration is a simple and effective method of sperm retrieval, which can be performed from the same testis up to several times with good recovery of motile spermatozoa for ICSI and maintaining high fertilization and pregnancy rates, in men with both obstructive and nonobstructive azoospermia. PMID- 11411435 TI - Severe intraabdominal bleeding after transvaginal oocyte retrieval for IVF-ET and coagulation factor XI deficiency: a case report. PMID- 11411436 TI - Oocyte donation treats infertility in survivors of malignancies: ten-year experience. AB - Women who are survivors of malignancy may achieve pregnancy through oocyte donation. The largest obstacle to successful reproduction in cancer patients relates to iatrogenic damage to the primary reproductive organs associated with their primary oncology treatment. Reactive damage from surgery, radiation, or chemotherapy may render these organs nonfunctional and irreparable. In cases where the ovary is primarily affected, oocyte donation provides a logical alternative for childbearing. PMID- 11411437 TI - Estimating and testing mediation and moderation in within-subject designs. AB - Analyses designed to detect mediation and moderation of treatment effects are increasingly prevalent in research in psychology. The mediation question concerns the processes that produce a treatment effect. The moderation question concerns factors that affect the magnitude of that effect. Although analytic procedures have been reasonably well worked out in the case in which the treatment varies between participants, no systematic procedures for examining mediation and moderation have been developed in the case in which the treatment varies within participants. The authors present an analytic approach to these issues using ordinary least squares estimation. PMID- 11411438 TI - Review of assumptions and problems in the appropriate conceptualization of effect size. AB - Estimation of the effect size parameter, D, the standardized difference between population means, is sensitive to heterogeneity of variance (heteroscedasticity), which seems to abound in psychological data. Pooling s2s assumes homoscedasticity, as do methods for constructing a confidence interval for D, estimating D from t or analysis of variance results, formulas that adjust estimates for inflation by main effects or covariates, and the Q statistic. The common language effect size statistic as an estimate of Pr(X1 > X2), the probability that a randomly sampled member of Population 1 will outscore a randomly sampled member of Population 2, also assumes normality and homoscedasticity. Various proposed solutions are reviewed, including measures that do not make these assumptions, such as the probability of superiority estimate of Pr(X1 > X2). Ways to reconceptualize effect size when treatments may affect moments such as the variance are also discussed. PMID- 11411439 TI - The converse inequality argument against tests of statistical significance. AB - Critics have put forth several arguments against the use of tests of statistical significance (TOSSes). Among these, the converse inequality argument stands out but remains sketchy, as does criticism of it. The argument states that we want P(H/D) (where H and D represent hypothesis and data, respectively), we get P(D/H), and the 2 do not equal one another. Each of the terms in 'P(D/H) not equal to P(H/D)' requires clarification. Furthermore, the argument as a whole allows for multiple interpretations. If the argument questions the logic of TOSSes, then defenses of TOSSes fall into 2 distinct types. Clarification and analysis of the argument suggest more moderate conclusions than previously offered by friends and critics of TOSSes. Furthermore, the general method of clarification through formalization may offer a way out of the current impasse. PMID- 11411440 TI - Meta-analysis of correlation coefficients: a Monte Carlo comparison of fixed- and random-effects methods. AB - The efficacy of the Hedges and colleagues, Rosenthal-Rubin, and Hunter-Schmidt methods for combining correlation coefficients was tested for cases in which population effect sizes were both fixed and variable. After a brief tutorial on these meta-analytic methods, the author presents two Monte Carlo simulations that compare these methods for cases in which the number of studies in the meta analysis and the average sample size of studies were varied. In the fixed case the methods produced comparable estimates of the average effect size; however, the Hunter-Schmidt method failed to control the Type I error rate for the associated significance tests. In the variable case, for both the Hedges and colleagues and Hunter-Schmidt methods, Type I error rates were not controlled for meta-analyses including 15 or fewer studies and the probability of detecting small effects was less than .3. Some practical recommendations are made about the use of meta-analysis. PMID- 11411441 TI - The effect of ignoring item interactions on the estimated discrimination parameters in item response theory. AB - Most item response theory models assume conditional independence, and it is known that interactions between items affect the estimated item discrimination. In this article, this effect is further investigated from a theoretical perspective and by means of simulation studies. To this end, a parametric model for item interactions is introduced. Next, it is shown that ignoring a positive interaction results in an overestimation of the discrimination parameter in the two-parameter logistic model (2PLM), whereas ignoring a negative interaction leads to an underestimation of the parameter. Furthermore, it is demonstrated that in some cases the item characteristic curves of the 2PLM and of an item involved in an interaction are quite similar, indicating that the 2PLM can provide a good fit to data with interactions. PMID- 11411442 TI - Multidisciplinary case conference reviews: improving outcomes for nursing home residents, carers and health professionals. AB - AIM OF STUDY: This study sought to determine whether multidisciplinary case conference reviews improved outcomes for nursing home residents, and the effects of this team approach to resident care on carers, including the hands-on carers employed by the nursing home, and health professionals. METHOD: 245 residents of three Canberra nursing homes were enrolled in this non-randomised controlled trial. The intervention consisted of sessions of three case conference reviews held between 10/4/96 and 4/12/96. These sessions were attended by the General Practitioners (GPs) of the residents discussed, the GP project officer from the ACT Division of General Practice, a clinical pharmacist, senior nursing staff, other health professionals e.g. physiotherapist, and occasionally the resident concerned or their representative. At each review, a case presentation by the resident's GP was followed by a multidisciplinary discussion of all aspects, medical and non-medical, of the resident's care. The review concluded with a management plan for the resident. In total 75 residents were reviewed. MAIN OUTCOME MEASURES: Medication use and cost, and mortality. RESULTS: One month after the reviews were completed comparisons between those who were reviewed and those who were not showed non-significant reductions in medication orders, medication cost, and mortality in the reviewed group. Many of the 92 recommendations in the management plans that were carried out benefited the residents (n = 37) and/or carers (n = 24). The responses of the GPs and the Directors of Nursing to the reviews were overwhelmingly positive. CONCLUSION: Recommendations arising from multidisciplinary case conferences were carried out to the benefit of patients and carers. Given the support shown by key stakeholders, multidisciplinary conferences should be used more. PMID- 11411443 TI - Antidepressant prescription for depression in general practice in The Netherlands. AB - OBJECTIVE: To evaluate diagnoses and the prescription of antidepressants drugs for depression, based on data from 1993 to 1998 related to consultations for depression in general practice in the Netherlands. METHOD: An exploratory analysis of data provided by IMS Health. Consecutive annual representative samples of 640 GPs regularly registered data on morbidity and (drug) treatment for specific medical problems in special booklets for a period of one week. The data refer to consultations for depression. The diagnoses were initially based on ICD-9 criteria and later ICD-10 criteria for depression. MAIN OUTCOME MEASURES: Number of diagnoses of depression and number of prescriptions for antidepressants. RESULTS: The number of first consultations for depression rose gradually, from 909 in 1993 (95% CI: 774 to 1043), to 1,482 (CI: 1292 to 1672) in 1998: an increase of 63%. For an individual GP, this represents an increase from 18 to 30 in first consultations for depression. The number of prescriptions for antidepressants also increased, mainly during repeat consultations: a 278% increase, from 3,708 in 1993 to 14,024 in 1998 (CIs: 3,504 to 3,912 and 13,252 to 14,796). Antidepressants were prescribed during the first consultation 564 times in 1993 compared with 1,080 times in 1998. The first contact with a GP for depression led to an antidepressant prescription in 62% of cases in 1993 and 73% in 1998. Although the guidelines issued by the Dutch College of General Practitioners in 1994 recommended tricyclic antidepressants (TCAs) for the treatment of depression, paroxetin, fluoxetin, and mirtazapin (SSRIs) accounted for 56% of the prescribed antidepressants. CONCLUSION: Between 1993 and 1998, GPs in the Netherlands prescribed increasingly more antidepressants, and they prescribed more SSRIs than TCAs. Furthermore, GPs diagnose depression more frequently and the number of repeat consultations has increased. PMID- 11411444 TI - Validity of questions in the use of specific drug-groups in health surveys. AB - OBJECTIVE: The aim of this study was to investigate whether morbidity in the general population could be assessed by questions on drug use in the Norwegian Health Survey 1995. MATERIAL AND METHOD: A sample of 6,702 persons, aged 20-79 years was interviewed in their homes using computer-assisted personal interviewing (CAPI). MEAN OUTCOME MEASURE: The validity of questions on use of analgesics and drugs against dyspepsia/peptic ulcer has been assessed according to categories of self-evaluated health. RESULTS: There was a difference between sporadic and daily users of the drugs to what extent they rated their health as poor. The validity of the drug questions assessed by sensitivity and specificity, showed that only using a dichotomous outcome variable, is too low to give a sufficiently valid measure of the morbidity in the population. CONCLUSIONS: Using "yes" or "no" as the only outcome of drug questions has the unfortunate effect of putting together chronic users of drugs with infrequent users for all of the subsequent analyses, which results in a considerable measurement error. This implies a need for improved methods to determine the optimal recall period for different drugs and it is crucial to include more details in questions on drug use to increase the validity of this information. PMID- 11411445 TI - Self reported clinical pharmacist interventions under-estimate their input to patient care. AB - OBJECTIVE: Pharmacists' impact on individual patient care is difficult to measure especially the contribution made by clinical pharmacy ward visits. This study set out to determine what activities pharmacists actually undertook on a clinical pharmacy ward visit and compare this with the usual method of measuring clinical pharmacist performance, self-reported pharmacist interventions. METHOD: Observational analysis was carried out on 16 pharmacists providing a ward clinical pharmacy service in four acute hospitals. MAIN OUTCOME MEASURE: Percentage of pharmacist interventions recorded. RESULTS: A total of 34 wards were visited during the study which included both medical and surgical specialties. Average time spent per patient was less than two minutes for most pharmacists and three-quarters of the pharmacists checked over 80% of patient drug charts. Interventions represented 68% of pharmacist activities on the wards but on questioning the pharmacists reported that they would record only 31% of those interventions. Comparison of the interventions the pharmacists stated they would record with an historical sample of recorded interventions from the four hospitals showed a similar pattern in each intervention category. However, pharmacists were more likely to record interaction type interventions and less likely to record interventions on incomplete/illegal prescriptions, which were regarded as routine practice. CONCLUSION: Pharmacists reported they would record less than one-third of interventions observed. If recorded pharmacist interventions continue to be used the main source of evidence of outputs of clinical pharmacy service, a better way of capturing this data needs to be developed. PMID- 11411446 TI - Patient-perceived problems, compliance, and the outcome of hypertension treatment. AB - OBJECTIVE: To study the associations between the outcome of antihypertensive therapy with both patient-perceived problems and patient initiated modification of dosage instructions. DESIGN AND METHODS: In this cross-sectional survey, all chronic hypertensives aged less than 75 years (n = 971) visiting nine Finnish pharmacies between May and September of 1996 were asked to participate. Of the 866 agreeing to participate, 482 returned the questionnaire (56%). After excluding persons with missing data, the final study population consisted of 428 hypertensive patients. Information on problems with treatment, the modification of dosage instructions, and blood pressure levels was based on patient self reports. RESULTS: Two-thirds (68%) of the study population reported suffering from one or more problems. The most common problems were symptoms of high blood pressure and adverse drug effects. Thirty-one percent of the male respondents and 21% of the female respondents reported having modified their dosage instructions. Only 36% of the patients had reached the goal blood pressure (< 160/90 mmHg). Patients having problems with hypertension treatment were significantly more likely to have modified their dosage instructions than those without problems (3+ problems, adjusted OR = 4.8). Not reaching goal blood pressure levels was significantly associated with both high number of patient-perceived problems (3+ problems, adjusted OR = 2.1) and modification of dosage instructions (adjusted OR = 1.9). CONCLUSION: The poor outcome in antihypertensive therapy is associated with both patient-perceived problems and patient initiated modification of dosage instructions. PMID- 11411447 TI - The impact of the pharmacist on an elective general surgery pre-admission clinic. AB - OBJECTIVE: A traditional clinical pharmacy service for surgical patients involves a ward visit that identifies and responds to issues after prescribing has taken place. It was hypothesised that pharmacist involvement in a general surgery pre admission clinic (PAC) would provide elective patients with significantly better pharmaceutical care than ward visits alone. METHOD: 100 elective general surgery patients were recruited into 2 groups as part of a non-randomised controlled trial. The groups were comparable in terms of age, surgical procedure and use of medicines. The control group received standard ward visits. The intervention group had pharmaceutical assessment in a pre-admission clinic. This assessment incorporated pharmacist-led drug history taking and prescription transcription. OUTCOME MEASURES: Number, classification and clinical significance of pharmaceutical interventions. RESULTS: A pharmacist made 76 interventions in the PAC group and 79 interventions in the ward group. No prescribing errors and omissions were made in the PAC group. The clinical significance of PAC interventions was rated more highly than ward interventions on two scales. Involvement in the PAC increased the pharmacist's workload, but saved time for surgical and nursing staff. Surgical and nursing staff identified writing discharge prescriptions as the most valuable service provided by the PAC pharmacist. CONCLUSION: By ensuring safe prescribing in the PAC group, the pharmacist was able to become more involved in clinical issues. Pharmacist involvement in PACs presents a useful opportunity for expansion of pharmacy services. However, extra resources would be needed in order to provide these services fully. PMID- 11411448 TI - Determinants of change in the intention to use benzodiazepines. AB - OBJECTIVE: To examine attitudes and beliefs associated with changes in the intention to use benzodiazepines during the six-month period after first benzodiazepine use. DESIGN: Population-based 6-month follow-up with 3 measurement points (baseline, 2 weeks after inclusion, 6 months after inclusion). SETTING: Starting or initial benzodiazepine users were included during a period of 4 months from November 1994 in the only pharmacy of a Dutch community of 13,500 people. MEASURES: Variables proposed by the Model of Planned Behaviour and the Health Belief Model. Drug exposure data from automated pharmacy records. RESULTS: At baseline, the intention to use benzodiazepines was primarily predicted by the perceived norm of the general practitioner regarding benzodiazepine use, and by the participants' own attitudes. After fourteen days, the determinants of change in the intention to use benzodiazepines were the initially perceived norm of the general practitioner and the change in the severity of participants' illnesses. After six months, the change in the severity of the illness and the perceived health benefits of benzodiazepines at the time of inclusion were the main determinants of the change in the intention to use benzodiazepines between the second and third measurement point. The intention to use benzodiazepines showed a decrease during follow-up. The three intention measures were significant predictors of actual benzodiazepine use during the year following baseline assessment. CONCLUSION: The study sheds light on interesting determinants of decrease or increase in the intention to use during the six-month period after first benzodiazepine use. PMID- 11411449 TI - Evidence based information on drug use during pregnancy: a survey of community pharmacists in three countries. AB - OBJECTIVE: To evaluate whether community pharmacists provide evidence-based information to women inquiring about specific drug use during pregnancy. DESIGN: A trained female student posing as a surrogate shopper requested information about the relative safety/risks of medications during pregnancy in two scenarios. Forty randomly selected pharmacies were surveyed in the Netherlands, Canada and Iceland, and pharmacists' recommendations were noted. Main outcome measures included the type of information that was provided, its presentation, and the source of information used. RESULTS: A relatively small proportion of pharmacists surveyed, provided evidence-based information regarding the drugs in question. Only 14% referred to current medical literature, while 60% consulted the product monograph. Over 90% of pharmacists referred the client to a physician. CONCLUSIONS: Community pharmacists do not disseminate evidence-based recommendations when counseling women on drug use in pregnancy, and need further education on resources concerning drugs in pregnancy that are currently available. PMID- 11411450 TI - Sharp transients in the EEGs of non-epileptic adult patients receiving sevoflurane. AB - OBJECTIVE: In this article unexpected EEG findings are described which were observed during EEG monitoring under sevoflurane anesthesia. METHOD: In seven non epileptic adult patients sevoflurane was administered as inhalation anesthetic during routinely performed surgical operations. The EEG was recorded continuously as part of the standard monitoring process and served mainly as a dosage guide for anesthetics/narcotics. MAIN OUTCOME MEASURE: Occurrence of sharp transients in the EEG resembling distinctive waves which can be seen in epileptic disorders. RESULTS: In six of the seven patients under 8.0% sevoflurane, sharp transients were observed which appeared in very deep EEG stages, mostly with endtidal sevoflurane concentrations of 4.8-5.9%. The findings are in accordance with observations in non-epileptic children from our clinic. CONCLUSIONS: The clinical significance of the observed EEG pattern under sevoflurane anesthesia is still unclear. Taking into consideration that convulsive and nonconvulsive status epilepticus can be followed by signs of brain damage, it would appear to be important to further investigate the phenomenon. PMID- 11411451 TI - Submission bias. PMID- 11411452 TI - Guidelines for cytotoxic chemotherapy in adults. A document for local expert groups in the United Kingdom preparing chemotherapy policy documents. PMID- 11411453 TI - Open position for decision makers. PMID- 11411454 TI - Recombinant hCG (OVIDREL) and recombinant interferon-(beta)1a (REBIF). (No. 13 in a series of articles to promote a better understanding of the use of genetic engineering). PMID- 11411455 TI - Quiz case. Mesenteric panniculitis (sclerosing mesenteritis, mesenteric lipodystrophy, retractile or liposclerotic mesenteritis. PMID- 11411456 TI - ACOG committee opinion: Tamoxifen and endometrial cancer. PMID- 11411457 TI - ACOG committee opinion: Ethical dimensions of seeking and giving consultation. PMID- 11411458 TI - Poorly differentiated synovial sarcoma: a case report. AB - Poorly differentiated synovial sarcoma is a rare soft tissue tumor. We studied a case arising in the pleural cavity of a young subject, characterised by the presence of spindle cell, small cell, and large epithelioid cell areas. We performed stains for mucosubstances and analysed the expression of cytokeratins 5/6, 7, 8, 18, 19, CEA, CD34, Ber-Ep4 and calretinin to characterize the phenotype of this neoplasm. We furthermore assessed immunohistochemically the presence of p53, Bcl-2, Bax and caspase 3, four apoptotic markers, to evaluate a relationship between apoptotic activity and the behaviour of this tumor. Our findings showed a strong presence of calretinin, p53 and Bcl-2 in all three areas. The possibility that poorly differentiated synovial sarcoma could be calretinin-positive was a new data, to our knowledge, and it could be of some importance in diagnostic pathology. Moreover, the negligible positivity for Bax and caspase 3 suggested that the minor role of programmed cell death could be one of the causes of the aggressive behaviour of this tumor. These data also suggest that the reduction of apoptotic phenomena in poorly differentiated synovial sarcoma could be considered one of the major mechanisms of tumoral growth. PMID- 11411459 TI - Relaxing on the run: stress management for the busy professional. PMID- 11411460 TI - [Quid? Cancer of the left colonic angle with colonic-splenic fistula, thrombosis of the splenic vein and left portal branch]. PMID- 11411461 TI - Stress hormone dysregulation at rest and after serotonergic stimulation among alcohol-dependent men with extended abstinence and controls. AB - BACKGROUND: Alcohol dependence has been associated with long-lasting alterations in limbic-hypothalamic-pituitary-adrenal (LHPA) axis and serotonin (5 hydroxytryptamine [5-HT]) function. Other conditions that are associated with alcoholism (cigarette smoking and antisocial personality disorder [ASPD]) have been linked with disturbances in these interrelated systems. We evaluated the stress hormone response to 5-HTergic stimulation in alcohol-dependent men with extended abstinence (average abstinence duration, 4.3 months) and controls to determine the relative contributions of alcoholism, cigarette smoking, and ASPD on baseline and provoked plasma cortisol and adrenocorticotropin hormone (ACTH) concentrations. METHODS: One hundred nine alcohol-abstinent men with alcohol dependence (62%), habitual smoking (70%), and ASPD (43%) received D,L fenfluramine (100 mg po) in a randomized, double-blind, placebo-controlled, crossover trial. The group of recovering alcohol-dependent individuals included abstinent primary alcohol-dependent men and alcohol-dependent men with ASPD, whereas the group of non-alcohol-dependent men comprised healthy controls and non alcohol-dependent men with ASPD. Plasma cortisol and ACTH levels were obtained at AM baseline and at half-hour intervals after drug administration. Subjective ratings of drug response and physiological measures were also obtained at baseline and every 30 min. RESULTS: Abstinent alcohol-dependent men had significantly lower (approximately 20%) AM baseline plasma cortisol concentrations than non-alcohol-dependent men on both challenge days; however, no differences between the groups were observed with regard to resting AM plasma ACTH levels. After adjusting for these baseline differences, recovering alcohol dependent men (area under curve = 35.6 +/- 37.4 [microg/dl] x min) had a twofold greater cortisol response to fenfluramine than non-alcohol-dependent men (area under curve = 17.5 +/- 32.5 [microg/dl] x min) (F = 5.1; df = 1,105; p < 0.03). The elevated cortisol response, which occurred primarily along the descending limb of the response curve, was paralleled by a nonsignificant statistical trend for alcohol-dependent men to also exhibit a greater ACTH response to fenfluramine at the 210-min (p < 0.07) and 240-min (P < 0.09) time points as compared with non alcohol-dependent men. Cigarette smoking and ASPD did not affect hormonal responses, nor could the groups' subjective ratings and physiological measures be distinguished. CONCLUSIONS: Alcohol-dependent men with extended abstinence differed from age- and race-matched non-alcohol-dependent men in resting AM and fenfluramine-induced plasma cortisol levels. This dysfunction in glucocorticoid homeostatic mechanisms was associated with alcoholism and not with smoking or ASPD. We also observed a nonsignificant statistical trend for plasma ACTH levels to be elevated among alcohol-dependent men along the descending limb of the response curve. Alcohol-dependent men seemed to have inherited or acquired damage to 5-HT-regulated LHPA axis function, the precise mechanisms and sites of which remain to be determined. PMID- 11411462 TI - Use of cultured cells in assessing ethanol toxicity and ethanol-related metabolism. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Terrence M. Donohue, Jr, and Dahn L. Clemens. The presentations were (1) Characterization of single and double recombinant hepatoma cells that express ethanol-metabolizing enzymes, by Terrence M. Donohue, Jr; (2) Inhibition of cell growth by ethanol metabolism, by Dahn L. Clemens; (3) Use of transfected HeLa cells to study the genesis of alcoholic fatty liver, by Andrea Galli and David Crabb; (4) CYP2E1-mediated oxidative stress induces COL1A2 mRNA in hepatic stellate cells and in a coculture system of HepG2 and stellate cells, by Natalia Nieto; (5) Transforming growth factor-alpha secreted from ethanol-exposed hepatocytes contributes to development of alcoholic hepatic fibrosis, by Junji Kato; and (6) Effect of ethanol on Fas-dependent caspase-3 activation and apoptosis in CD4+ T cells, by Shirish S. Barve. PMID- 11411463 TI - Selected bibliography. PMID- 11411464 TI - Retraction. PMID- 11411465 TI - Erratum: a quantification of the lung surface area in emphysema using computed tomography. PMID- 11411466 TI - Aging, immunity, and cancer. PMID- 11411467 TI - Participation deadline extended 10 weeks for Medicare managed care organizations. PMID- 11411468 TI - Christopher Reeve and 7 scientists sue Bush administration over lagging stem cell funds. PMID- 11411469 TI - Retirees now less likely to have employer-based insurance. PMID- 11411470 TI - Effect of age, gender, and repeated measures on intraoral air pressure in normal adults. AB - The effect of age, gender, and repeated measures on intraoral air pressure (P0) was examined. Sixty adults comprised of 10 males and 10 females in each of three age groups (i.e., 20-39, 40-59, and 60-83 years) participated. P0 was assessed during voiceless stop plosive /p/ productions in repeated vowel/consonant syllables. The three medial plosives of a seven-syllable train were averaged to comprise a token. Five tokens were obtained and averaged for each of three trials. Thus each participant contributed 105 syllables and a subsequent three P0s for analyses. There was no statistically significant difference in P0 as a function of age or gender (P > 0.05). These findings support the conception that P0 remains stable throughout adulthood and is not dependent on gender. Differences in repeated measures of P0 attained statistical significance (P = 0.03), however the mean differences between trials (0.23 cm H20) were negligible and deemed to be clinically insignificant. Thus, across a short sampling session, P0 is a relatively stable measurement and does not change as a function of age or gender. PMID- 11411471 TI - Perception of pitch and roughness in vocal signals with subharmonics. AB - Pitch and roughness were rated according to the extent of amplitude modulation (AM) and frequency modulation (FM) of a subharmonic [fundamental frequency (F0)/2]. The objective was to determine the identification boundaries for pitch and roughness and to discover how both kinds of modulation affect these boundaries. Another objective was to judge the reliability between subjects when identifying subharmonic-related pitch and roughness. Three procedures were used: ABX comparisons, method of adjustment, and rating of roughness. Results indicated that the crossover point to the lower pitch (associated with the subharmonic) occurred between 10% and 30% modulation, depending on modulation type and F0. Subjects demonstrated highly variable perceptions of pitch and roughness, with poor intersubject reliability. PMID- 11411472 TI - Level and center frequency of the singer's formant. AB - The "singer's formant" is a prominent spectrum envelope peak near 3 kHz, typically found in voiced sounds produced by classical operatic singers. According to previous research, it is mainly a resonatory phenomenon produced by a clustering of formants 3, 4, and 5. Its level relative to the first formant peak varies depending on vowel, vocal loudness, and other factors. Its dependence on vowel formant frequencies is examined. Applying the acoustic theory of voice production, the level difference between the first and third formant is calulated for some standard vowels. The difference between observed and calculated levels is determined for various voices. It is found to vary considerably more between vowels sung by professional singers than by untrained voices. The center frequency of the singer's formant as determined from long-term spectrum analysis of commercial recordings is found to increase slightly with the pitch range of the voice classification. PMID- 11411473 TI - The effect of cricothyroid muscle action on the relation between subglottal pressure and fundamental frequency in an in vivo canine model. AB - The relation between subglottal pressure (Ps) and fundamental frequency (F0) in phonation was investigated with an in vivo canine model. Direct muscle stimulation was used in addition to brain stimulation. This allowed the Ps-F0 slope to be quantified in terms of cricothyroid muscle activity. Results showed that, for ranges of 0-2 mA constant current stimulation of the cricothyroid muscle, the Ps-F0 slope ranged from 10 Hz/kPa to 60 Hz/kPa. These results were compared to similar slopes obtained in a previous study on excised larynges in which the vocal fold length was varied instead of cricothyroid activation. A physical interpretation of the Ps-F0 slope is that the amplitude-to-length ratio of the vocal folds decreases with CT activity, resulting in a smaller time varying stiffness. In other words, there is less dependence of F0 on amplitude of vibration when the vocal folds are long instead of short. PMID- 11411474 TI - Multimodal standardization of voice among four multicultural populations: fundamental frequency and spectral characteristics. AB - A stratified random sample of 20 males and 20 females matched for physiological factors and cultural-linguistic markers were examined to determine differences in fundamental frequency and spectral characteristics during prolongation of three vowels: [a], [i], and [u]. The ethnic-gender breakdown included four sets of five male and five female subjects comprised of Caucasian and African-American speakers of standard American English, native Hindi Indian speakers, and native Mandarin Chinese speakers. Acoustic measures were analyzed using the Multidimensional Voice Program (Kay Elemetrics, Lincoln Park, NJ) (Model 4305) from which fundamental frequency and associated acoustic spectra were extracted from a 200-ms sample of each vowel token. Statistically significant group differences for the main effects of culture, race, and gender were found. The acoustic differences found are attributed to biomechanical, physiological, cultural, and linguistic factors. PMID- 11411475 TI - Devising an objective nasal vibration test for nasal resonatory disorders. AB - The present study investigates the clinical applicability of a new device, which objectively measured nasal resonating vibration via piezoelectric vibratory sensor in 10 normal volunteers, 10 patients with definite hypernasality, and 10 nasal polyposis patients. For the assessment of the hypernasality, the ratio of /ng/ to /a/ as well as the ratio of /mama/ to /papa/ passages were used. For the evaluation of hyponasality, the ratio of nasal vibration postinduced to preinduced cul-de-sac resonation was calculated. The control group showed the ratio of /ng/ to /a/ and /mama/ to /papa/ passages to be larger than 8, whereas the ratio was markedly lower in the hypernasality group. The vibratory signals of /a/ and /ng/ increased markedly in the control and the hypernasality groups after inducing cul-de-sac resonation, but the change was minimal in the hyponasality group. This new device could detect nasal resonatory disorders and readily differentiate between hypernasality and hyponasality. PMID- 11411476 TI - Use of speech recognition software: a vocal endurance test for the new millennium? AB - Speech recognition software for the personal or office computer is a relatively new area of technology. As the number of these products has increased so has use of this software. Some individuals will employ speech recognition systems due to difficulty with the conventional keyboard and mouse interface: others will use it for perceived efficiency or simply novelty. Regardless of the reason for use of this technology, the voice demands associated with extended or frequent use can be high, placing the user at risk for vocal difficulties. This paper reviews the case of an individual referred to our multidisciplinary voice care program for evaluation and treatment of vocal difficulties that began secondary to utilization of speech recognition software. We discuss medical and vocal histories, examination findings, treatment, and treatment outcomes. PMID- 11411477 TI - Preparation of the speech-language pathologist specializing in voice: an educational survey. AB - This study investigates the academic and clinical preparation a speech-language pathologist receives in the area of voice. The purpose is to determine how to structure continuing education and specialty training in vocology, defined as the science and practice of voice habilitation. Surveys were sent to the graduate programs in speech-language pathology accredited by the American Speech-Language Hearing Association (ASHA) in 1994, and again in 1999. Results from the 1994 survey indicated that students received limited information and clinical experience in handling typical voice cases. Although most programs required a voice disorders course, fewer mandated coursework in normal voice production or clinical experience with voice patients. The follow-up survey showed that the educational environment in 1999 is similar to that in 1994. It is evident that the programs are in compliance with the Educational Training Board model developed by ASHA for basic clinical competence, but no structure for specialty training has yet emerged. PMID- 11411478 TI - Plasticity of voice quality: a prognostic factor for outcome of voice therapy? AB - Plasticity of voice quality is defined here as the degree of improvement in deviant voice quality that can be achieved immediately or quasi-immediately by changing basic voicing conditions, posture, articulation or resonance, breathing mechanics, laryngeal position, or auditory feedback. Thirty-two adult patients with various benign organic voice pathologies, and who had a (preoperative) functional voice therapy, were scored before therapy using a weighted multidimensional Index of Voice Plasticity (IVP). The hypothesis is that IVP could be a predictor of the final outcome of functional voice therapy, and therefore a correlation with a comparable quantification of the actual results of the therapy was investigated. The IVP shows a satisfactory correlation (Spearman's rho = 0.68) with the efficacy of (preoperative) voice therapy. The IVP also significantly differs between diagnostic categories. Although its predictive value remains limited, the Index of Voice Plasticity seems helpful in decision making for indication of (presurgical) voice therapy. PMID- 11411479 TI - Performance anxiety: loss of the spoken edge. AB - The study aimed to assess whether a specific training program in vocal and physical skills could reduce the level of perceived performance anxiety. Eighteen undergraduate performing arts students were divided into two even groups. The experimental group experienced 10 two-hour sessions in a specialized vocal and physical skills training program. The control group received the same number of sessions in text analysis by the same teacher. Ten weeks after the training period, both groups were videotaped delivering a short speech of general interest. The videotaped material was assessed by four judges, using a visual analog scale (VAS). Perceptual variables considered were eight vocal and physical features: physical ease, physical presence, effective gesture use, effective eye contact, correct breath use, suitable pace, vocal variety, speech clarity, and one performance feature: perceived performance anxiety. Results indicated that the particular vocal and physical skills training program used showed positive results in effectively reducing the level of perceived performance anxiety. The experimental group scored significantly higher on each of the eight vocal and physical features and significantly lower on the perceived anxiety feature than the control group. This research has implications for those affected by performance anxiety in vocal presentation. PMID- 11411480 TI - The relationship between dysphonia and sinusitis: a pilot study. AB - Though some clinicians believe chronic sinusitis is associated with dysphonia, studies confirming such a relationship are not available. This pilot study investigated vocal characteristics of chronic sinusitis patients compared to those of nonsinusitis patients. Nineteen male patients were evaluated in a single blind, nonrandomized, prospective study at a Department of Veterans Affairs Medical Center. After candidates for the experimental and control groups were thoroughly screened [nasal and laryngology examinations, sinus computed tomography (CT) scan, history], 10 chronic sinusitis patients were assigned to the experimental group, nine patients without chronic sinusitis were assigned to the control group. Visi-Pitch analysis of their speech evaluated fundamental frequency, intensity, and perturbation. The evaluator described voice quality and vowel prolongation. Videostroboscopic examination evaluated glottic closure, supraglottic activity, vocal fold edge, amplitude, mucosal wave, and nonvibrating portion. Visi-Pitch analysis and vowel prolongation for each group was compared using an independent t-test. The average fundamental frequency mean was 113.43+/ 32.75 Hz for the control group and 112.29+/-19.71 for the experimental group. This study established methods and parameters to determine if a relationship between chronic sinusitis and dysphonia actually exists. It did not establish a relationship between these entities, possibly due to the small sample size enrolled in the study, but determined that 126 subjects would be necessary to achieve significant results. PMID- 11411481 TI - Clinical analysis of voice change as a parameter of premenstrual syndrome. AB - We have evaluated the relationship between voice change and premenstrual syndrome (PMS) by comparing acoustic measurements made during the follicular phase and the premenstrual phase. Twenty-eight women were followed for 2 months for this study. Each participant was asked to produce an /a/ sound for 5 seconds at the midfollicular phase of the menstrual cycle and then 2-3 days before menstruation. Each voice sample was stored and analyzed by the Dr. Speech Science program. The voice data collected from all subjects during the two phases were compared. After that, the subjects were divided into a PMS-positive and PMS-negative group according to the criteria cited in the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV); the voice data from each group were compared separately between the two phases. There was no significant difference in the acoustic parameters between the two phases in all subjects (N = 28). In the PMS positive group (N = 16), jitter was significantly increased during the premenstrual phase compared to the follicular phase (p = 0.048). The patient's PMS score was not correlated with the severity of voice change. We conclude that the change of voice parameter was objectively identified in the PMS-positive group, therefore more careful voice habituation is required during the premenstrual phase in that group. PMID- 11411482 TI - Aerodynamic and acoustic characteristics of voice in Prader-Willi syndrome. AB - Prader-Willi syndrome (PWS) is a multisystem disorder caused by DNA abnormalities involving chromosome 15. Major characteristics are infant hypotonia, hypogonadism, mental retardation, a short stature, atypical facial appearance, and the onset of obesity due to insatiable hunger in early childhood. Also, speech and language abnormalities have been reported including voice disorders. These have seldom been studied in detail, however. This paper reports the results of an acoustic and aerodynamic investigation of the voice in 22 individuals with PWS. Two age groups were distinguished, a group of children [chronological age (CA) 6 years, 7 months through 11 years, 7 months; total intelligence quotient (TIQ) 40-88] and a group of adolescents and adults (CA 17 years, 1 month through 29 years, 5 months; TIQ 41-94). Both aerodynamic and acoustic parameters were obtained and compared with normative data from the Belgian Study Group on Voice Disorders. It was found that voice difficulties do commonly occur in individuals with PWS including impairment of frequency levels, voice quality, and poor aerodynamic capabilities. PMID- 11411483 TI - Laryngeal manifestations of postpoliomyelitis syndrome. AB - Postpoliomyelitis syndrome (PPS) is a disease that may occur in survivors of acute poliomyelitis several decades after their initial infection. It can present as dysphonia, with vocal weakness and fatigue. Swallowing, respiratory, and other laryngopharyngeal symptoms may be manifestations of the disease or they may represent worsening of previously stable and compensated deficits. Three cases of laryngeal changes in PPS with videostroboscopic and laryngeal electromyography findings highlight the features of this disorder. We review possible etiologies of laryngeal PPS, diagnostic criteria, and treatment, as well as the current literature. PMID- 11411484 TI - Management of vocal fold scar with autologous fat implantation: perceptual results. AB - Vocal fold scar disrupts the mucosal wave and interferes with glottic closure. Treatment involves a multidisciplinary approach that includes voice therapy, medical management, and sometimes surgery. We reviewed the records of the first eight patients who underwent autologous fat implantation for vocal fold scar. Information on the etiology of scar, physical findings, and prior interventions were collected. Videotapes of videostroboscopic findings and perceptual voice ratings [Grade, Roughness, Breathiness, Asthenia, Strain (GRBAS)] were randomized and analyzed independently by four blinded observers. Etiology of scar included mass excision (7), vocal fold stripping (3), congenital sulcus (2), and hemorrhage (1). Prior surgical procedures performed included thyroplasty (1), autologous fat injection (9), excision of scar (2), and lysis of adhesions (2). Strobovideolaryngoscopy: Statistically significant improvement was found in glottic closure, mucosal wave, and stiffness (P = 0.05). Perceptual ratings (GRBAS): Statistically significant improvement was found in all five parameters, including overall Grade, Roughness, Breathiness, Asthenia, and Strain (P = 0.05). Patients appear to have improved vocal fold function and quality of voice after autologous fat implantation in the vocal fold. Autologous fat implantation is an important adjunctive procedure in the management of vocal fold scar, and a useful addition to the armamentarium of the experienced phonomicrosurgeon. PMID- 11411485 TI - Video-assisted rigid endoscopic laryngosurgery: application to cases with difficult laryngeal exposure. AB - In some cases of microlaryngosurgery, laryngeal exposure with a direct laryngoscope is difficult because of a variety of reasons. In such cases, we now use a long rigid endoscope inserted into the side tube of the direct laryngoscope together with a video system. The lesion can be removed while the phonosurgeon observes the magnified images of the larynx on the video monitor. We describe the surgical procedure together with the preoperative and postoperative evaluation of vocal function in 13 patients with benign laryngeal lesions. A good surgical and vocal outcome was achieved in all cases. The method appeared to be useful for treating dysphonia in patients in whom it was difficult to expose the larynx with a direct laryngoscope. PMID- 11411486 TI - Water resources programs under the ax. PMID- 11411487 TI - Directions to "eureka!". PMID- 11411488 TI - Health consequences of the Chornobyl accident. PMID- 11411489 TI - Many citations support global warming trend. PMID- 11411490 TI - The scope of Medieval warming. PMID- 11411491 TI - A cross-protection experiment in pigs vaccinated with Haemophilus parasuis serovars 2 and 5 bacterins, and evaluation of a bivalent vaccine under laboratory and field conditions. AB - Cross-protection between Haemophilus parasuis serovars 2 and 5 was examined in pigs using a bacterin based vaccine, and subsequently the safety and efficacy of a bivalent vaccine were evaluated. Upon intratracheal challenge of a serovar 2 or 5 strain, pigs immunized with a monovalent vaccine were protected against challenge with a homologous serovar strain, but not with a heterologous serovar strain. Immunization with a bivalent vaccine containing both serovars 2 and 5 bacterins conferred protection in pigs against lethal challenge with each of the serovar strains. A total of 86 pigs from two SPF herds were injected with the bivalent vaccine intramuscularly twice at a four-week interval. No adverse reactions following the vaccination were observed. On day 7 after the second vaccination, vaccinated and non-vaccinated control pigs from herd A were transferred to herd B, where Glasser's disease had broken out. Pigs in the control group developed clinical signs of the disease, and 6 of 8 (75%) pigs died until slaughter, in contrast with only 4 of 46 (9%) pigs in the vaccinated group. In herd C, where there was no outbreak of Glasser's disease, complement fixation antibody titer was raised only in the vaccinated group. A challenge experiment on days 20 and 79 after the second vaccination showed that only the vaccinated pigs were protected. From these findings, the safety and efficacy of the bivalent vaccine were confirmed under laboratory and field conditions. PMID- 11411492 TI - Norsalsolinol uptake into secretory vesicles via vesicular monoamine transporter and its secretion by membrane depolarization or purinoceptor stimulation in PC12 cells. AB - The intracellular dynamics of norsalsolinol, a neurotoxin candidate causing parkinsonism-like symptoms, in PC12 cells was studied. We found that dopamine and norsalsolinol are co-localized to secretory granule layer by sucrose density gradient in norsalsolinol-treated PC12 cells. The norsalsolinol was actively taken up into isolated secretory vesicle fraction from PC12 cells with a Km value of 41.5+/-6.8 microM. The uptake of 10 microM of norsalsolinol was sensitive to reserpine (1 microM), an inhibitor of vesicular dopamine transporter, and dopamine, an endogenous substrate, but insensitive to GBR-12909, an inhibitor of dopamine transporter on plasma membrane. In norsalsolinol-treated PC12 cells, exposure to high K+ or ATP resulted in simultaneous release of norsalsolinol and dopamine. Time course of a release of dopamine and that of norsalsolinol evoked by 50 mM KCl or 100 microM ATP corresponded to each other. These releases were dependent on the concentrations of secretagogues. These data suggest that norsalsolinol is taken up with dopamine into secretory vesicle via vesicular catecholamine transporter. PMID- 11411493 TI - Capsaicin-induced relaxation in rabbit coronary artery. AB - In the present study mechanism of inhibitory effects of capsaicin on the contractility of rabbit coronary artery were studied by measurement of isometric tension and intracellular Ca2+ concentration. Capsaicin (1 microM to 30 microM) relaxed the coronary artery pre-contracted with prostaglandin (PG) F2alpha (1 microM) in a concentration-dependent manner. The PGF2alpha-induced increase in intracellular Ca2+ concentration was also inhibited. The effects of capsaicin were readily reversed by washing capsaicin from the bath. Capsaicin-induced relaxation was not attenuated by pretreatment with capsazepine (1 microM), a blocker of vanilloid receptor or ruthenium red (1 microM), a blocker of non selective cation channel. Previous exposure to a high concentration of capsaicin (100 microM) or repeated application of capsaicin did not eliminate the relaxation response to subsequent application of capsaicin. Increasing the external K+ concentration to 80 mM significantly attenuated the capsaicin-induced relaxation with simultaneous change in intracellular Ca2+ concentration. Pretreatment with iberiotoxin (100 nM), a blocker of Ca2+-activated K+ channel, only partially inhibited the capsaicin-induced relaxation. However, application of 4-aminopyridine (4-AP, 1 mM), a blocker of delayed rectifier K+ current significantly inhibited the capsaicin-induced relaxation with concomitant attenuation of the effect on intracellular Ca2+ concentration. These results indicate that capsaicin may have a direct relaxing effect on the smooth muscle contractility, and relaxation may be due to activation of the 4-AP-sensitive, delayed rectifier K+ channels in the rabbit coronary artery. PMID- 11411494 TI - Glycoconjugate in rat taste buds. AB - The taste buds of the fungiform papillae, circumvallate papilla, foliate papillae, soft palate and epiglottis of the rat oral cavity were examined by lectin histochemistry to elucidate the relationships between expression of glycoconjugates and innervation. Seven out of 21 lectins showed moderate to intense staining in at least more than one taste bud. They were succinylated wheat germ agglutinin (s-WGA). Dolichos biflorus agglutinin (DBA), Bandeiraea simplicifolia lectin-I (BSL-I), Ricinus communis agglutinin-I (RCA-I), peanut agglutinin (PNA), Ulex europaeus agglutinin-I (UEA-I) and Phaseolus vulgaris agglutinin-L (PHA-L). UEA-I and BSL-I showed moderate to intense staining in all of the taste buds examined. They strongly stained the taste buds of the epiglottis, which are innervated by the cranial nerve X. UEA-I intensely stained the taste buds of the fungiform papillae and soft palate, both of which are innervated by the cranial nerve VII. The taste buds of circumvallate papilla and foliate papillae were innervated by the cranial nerve IX and strongly stained by BSL-I. Thus, UEA-I and BSL-I binding glycoconjugates, probably alpha-linked fucose and alpha-D-galactose, respectively, might be specific for taste buds. Although the expression of these glycoconjugates would be related to the innervation of the cranial nerve X, the differential expression of alpha-linked fucose and alpha-D-galactose might be related to the innervation of the cranial nerve VII and IX, respectively. PMID- 11411495 TI - Determination of oral dosage and pharmacokinetic analysis of flecainide in horses. AB - To determine oral dosage and to evaluate the pharmacokinetics in horses of orally administered flecainide, an antiarrhythmic drug, the correlations between its plasma concentration and PR, QRS and QT intervals in equine electrocardiograms (ECG) were investigated. Six healthy horses were administered a randomly ordered dose of 4 or 6 mg/kg of flecainide acetate. The ECG was monitored (heart rate (HR), PR, QRS, and QT intervals) and blood was taken at timed intervals to measure the plasma flecainide concentrations pre- and post-administration. The maximum plasma concentration reached 1014+/-285 (SD) ng/m/ in 45+/-13 min and 1301+/-400 ng/ m/l in 60+/-37 min for doses of 4 and 6 mg/kg flecainide, respectively. From the pharmacokinetic analysis, clearance rates were 14.6+/-6.4 and 11.7+/-5.2 ml/kg/min and terminal elimination half-lives were 228+/-53 and 304+/-87 min. The QRS and QT intervals increased significantly for both doses following administration, though HR and PR intervals did not change. Plasma flecainide concentrations were significantly correlated with QRS (r=0.935, P<0.001) and QT intervals (r=0.753, P<0.001). In conclusion, plasma concentrations of flecainide for treating equine atrial fibrillation were obtained by oral administration of 4 and 6 mg/kg, and the drug was rapidly eliminated from plasma in horses. PMID- 11411496 TI - Birth weight and gestation length of Japanese black calves following transfer of embryos produced in vitro with or without co-culture. AB - Birth weight and gestation length of calves following the transfer of in vitro produced (IVP) embryos with or without co-culture of cumulus cells, were compared to those produced in vivo (IVD). Spermatozoa from one Japanese Black bull were used for both IVP and IVD. IVP embryos were produced using two types of culture method: 1) co-culturing with cumulus cells in TCM 199 supplemented with calf serum (IVP-Co), and 2) non-co-culturing without cumulus cells in CR1aa supplemented with BSA / calf serum (IVP-NON-Co). Both IVP and IVD embryos were transferred non-surgically to Holstein recipients on day 7+/-1 of the estrous cycle. Birth weight and gestation length of half-sib single calves were analyzed. No differences were observed in birth weight and gestation length between IVP-Co and IVP-NON-Co calves (31.0 kg and 31.8 kg, and 291.9 days and 291.0 days, respectively). However, the birth weight of the IVP-Co and IVP-NON-Co calves was significantly higher than that of the IVD calves (P<0.01). Gestation length of the IVP-Co and IVP-NON-Co calves was also significantly longer than that of the IVD calves (P<0.01). PMID- 11411497 TI - Investigation of the short-term effect of chemonucleolysis with chondroitinase ABC. AB - Chondroitinase ABC (C-ABC) is expected to be a novel agent for chemonucleolysis. The effect of C-ABC was investigated by magnetic resonance (MR) and radiograph. C ABC was administered into the lumbar intervertebral disks on the clinically normal beagles (n=5), in a dose of 50 microl (12.5 units as C-ABC). MR scans were performed pre-dose, and 1, 3, 7, 14 and 28 days after administration of C-ABC, and the signal intensity (SI) of the nucleus pulposus was measured. Radiographs were taken pre-dose, and 1, 2, 3, 4, 5, 6, 7, 14 and 28 days post-dose, to evaluate narrowing of the disk space in terms of height index (HI). In addition, the quantity of the chondroitin sulfate (CS) and the hyaluronic acid (HA) in the nucleus pulposus were measured by high performance liquid chromatography on day 28 after dosing. SI and HI continuously decreased, following the injection to 37.1% and 78.9% of the pre-dose values, respectively. Statistically significant differences (p<0.01) were observed between the C-ABC group and the control group in the respects on day 1 post-dose. CS and HA contents of the nucleus pulposus were noted to be significantly decreased on day 28 (p<0.01) in the treated group. This agent proved to degenerate proteoglycans in the nucleus pulposus, thus progressively reducing the interdiskal pressure from day 1 post-dose onwards. It is concluded that C-ABC is expected to afford its efficacy from early in the course of chemonucleolysis. PMID- 11411498 TI - Time-related changes of the cardiovascular system during maintenance anesthesia with sevoflurane and isoflurane in horses. AB - To clarify time-related changes in equine cardiovascular system during maintenance anesthesia (180 min, 1.2 minimum alveolar concentration) with sevoflurane (Sev-group) compared to isoflurane (Iso-group) as the basis for clinical use of Sev, horses were examined for the heart rate (HR), mean arterial pressure (MAP), cardiac index (CI), systemic vascular resistance (SVR) and pre ejection period (PEP)/ejection time (ET) that is an index of the cardiac contractility. The HR was almost 30 beats/min in both groups without significant temporal change. MAP was significantly elevated with time but there was no significant difference between the groups. In the Sev-group, CI remained unchanged but the significant increase of CI with time was observed in the Iso group. In the Sev-group SVR was significantly higher than that of the Iso-group and increased with time. No significant difference of PEP/ET was seen between the groups, but PEP/ET lowered with time in the Iso-group in association with prolonged ET. The results indicated that the time-dependent elevation of MAP in the Sev-group reflected increased SVR without increase of CI and that it reflected increased CI resulting from increased stroke volume in the Iso-group in association with lowered PEP/ET, that is, increased cardiac contractility. PMID- 11411499 TI - Muscle architecture of the elongated nose in the Asian elephant (Elephas maximus). AB - The architecture of the M. caninus in the elongated nose was examined in the Asian elephant (Elephas maximus). The following complicated musculature of the M. caninus was observed in the proximal and distal regions of the nose: (1) Proximal region: In the superficial layer, the longitudinal bundles are confirmed in the dorsal part, and the obliquely-oriented ones in the ventral part. In the middle layer, some bundles run ventro-distally, while other ones represent longitudinally-oriented running. The deep layer consists of complicated architecture of many bundles. Some muscle bundles run medio-laterally, while the others extend proximo-distally in this space. (2) Distal region: In the dorsal part of the M. caninus, the bundles run at deep-superficial direction, while in the ventral part the bundles are longitudinally arranged. The bundles run at lateral direction near the septum of the nasal conduits. The N. facialis and N. infraorbitalis send many branches in the lateral area of the M. caninus in the trunk. This muscle architecture of multi-oriented bundles and well-developed innervation to them suggest that they enable the elongated nose to act as a refined manipulator in the Asian elephant. PMID- 11411500 TI - Neurotoxicity induced by a single oral dose of aniline in rats. AB - The neurotoxicity of aniline and its age-dependent responses were investigated in male rats. Groups of 6 rats, 4-week-old, were treated once with aniline (500, 750 or 1,000 mg/kg) or olive oil by gavage. Additional groups of 6 rats, 7- or 10 week-old, were treated once with 800 mg/kg of aniline or olive oil. Paralytic gait or hindlimb paralysis was observed between post-treatment days 8 and 15 in two out of six rats receiving 1,000 mg/kg of aniline at 4 weeks of age. On post treatment day 15, spongy change in the white matter of the spinal cord was observed in all rats receiving 750 or 1,000 mg/kg of aniline at 4 weeks of age. The lateral and ventral columns of the thoracic spinal cord were the most severely affected. Spongy change in the facial nerve and spinal trigeminal tracts of pons and medulla oblongata, and mild degeneration of the peripheral nerves was found in 3 out of 6 rats receiving 1,000 mg/kg of aniline. At the ultrastructural level, the spongy change was due to distention of the myelin sheath and splitting of the intraperiod line. Axons were well preserved in the affected nerve fibers. No abnormalities were seen in the neuronal cell bodies. Although transient cyanosis was observed in all rats receiving 800 mg/kg of aniline at 7- or 10-week old, as well as in rats receiving 750 or 1,000 mg/kg of aniline at 4-week-old, no treatment-related neurobehavioral or morphologic abnormalities were found in the former. These findings demonstrate the neurotoxicity of orally administered aniline for rats, depending upon the age of the animal at the time of administration. PMID- 11411501 TI - Immunohistochemical and ultrastructural study of ito cells (fat-storing cells) in response to extrahepatic bile duct ligation in broiler chickens. AB - The Ito cell (fat-storing cell) lies in perisinusoidal space of liver and has a variety of functions. We investigated the immunohistochemistry and ultrastructure of Ito cells in normal and cholestatic livers of broiler chickens. Immunohistochemistry demonstrated that Ito cells expressed HHF35 muscle actin, vimentin, desmin, glial fibrillary acidic protein (GFAP), neuron-specific enolase (NSE), chromogranin A and cytokeratins in normal livers. These cells were diffusely scattered throughout the lobules. Livers treated with extrahepatic bile duct ligation (BDL) showed cholestasis, fibrosis, proliferation of biliary ductules and Ito cells. The Ito cells were frequently found in fibrotic areas and were larger in size with more extensive immunoreactivity than those of normal livers. Ultrastructural study demonstrated that Ito cells were closely associated with the production of collagen fibers in BDL livers. These findings suggest that Ito cells actively react against hepatocytic injuries and play a major role in the hepatic fibrogenesis of cholestatic livers of chickens. PMID- 11411502 TI - Enzyme-linked immunosorbent assay for screening the plasma residues of tetracycline antibiotics in pigs. AB - The recommended therapeutic doses of three kinds of tetracyclines, oxytetracycline (OTC, withdrawal period, 10 days), chlortetracycline (CTC, withdrawal period, 5 days) and tetracycline (TC, withdrawal period, 5 days), were each administered to a group of 15 pigs. Blood was sampled before drug administration and during the withdrawal period. The concentration of tetracyclines in plasma, determined by semi-quantitative ELISA, was compared with that of internal standard (10 ppb as oxytetracycline). The absorbance ratio of internal standard to sample (B/Bs) was employed as an index to determine the tissue residues in pigs. All 45 plasma samples from nontreated pigs showed negative in the residue of any of three tetracycline antibiotics. OTC was detected in plasma of pigs treated until the 8th day, CTC until the 4th day, and TC was detected until the 3rd day of its withdrawal period. The present study showed that the semi-quantitative ELISA easily be adopted in predicting tissue residues for tetracycline antibiotics in live pigs. PMID- 11411503 TI - Supplemental embolization coil implantation for closure of patent ductus arteriosus in a beagle dog. AB - Embolization coil for the occlusion of patent ductus arteriosus (PDA) was inserted in a beagle dog diagnosed as PDA with systolic murmur. Residual shunt was observed three months postoperatively and then supplemental coil was inserted. In the past only one coil was required for the closure of PDA in our cases with good post-operative results. In this case, however, the largest coil presently available (8 mm in diameter with 5 loops) was inadequate to make sufficient closure within 3 months postoperatively and supplemental coil insertion was necessary to produce a successful occlusion. PMID- 11411504 TI - Comparison of partial ribosomal DNA sequences of Babesia gibsoni occurring in Miyazaki prefecture, Japan. AB - Nucleotide sequences of ribosomal DNA (rDNA) of Babesia (B.) gibsoni occurring in Miyazaki, western Japan, were examined using blood samples obtained from seven dogs suffering from natural canine babesiosis. DNA isolated from these blood samples was subjected to the polymerase chain reaction (PCR). The nucleotide sequences of the PCR products were determined and compared with other rDNA sequences of B. gibsoni isolated from Asia, Europe and U.S.A. Although homology values between our isolates and those isolated from Europe and U.S.A. were both 84.0%, respectively, our isolates were identical to the Asian types. In conclusion, B. gibsoni occurring in Miyazaki was revealed to have the genotype Asia 1 or Asia 2 from a comparison of the partial rDNA sequences. PMID- 11411505 TI - Effect of clindamycin therapy on phagocytic and oxidative activity profiles of spleen mononuclear cells in Babesia rodhaini-infected mice. AB - Spleen weight, the number of spleen mononuclear cells, and their phagocytic activity in groups of Babesia rodhaini-infected mice treated with diminazene diaceturate and clindamycin increased significantly in the early stage of treatment, and then decreased in the final stage of treatment to approximately the pre-infection level. The number of F4/80-positive macrophages and their oxidative activity per mean whole-spleen weight also increased significantly during the course of treatment in comparison with the untreated group. The increases in the clindamycin-treated group were more prominent than those in the group treated with diminazene diaceturate, suggesting the effectiveness of clindamycin therapy for murine babesiosis. PMID- 11411506 TI - Polymerase chain reaction-based restriction fragment length polymorphism pattern of porcine reproductive and respiratory syndrome virus directly from lung tissues without virus isolation in Korea. AB - Polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP) analysis was developed for directly typing porcine reproductive and respiratory syndrome virus (PRRSV) from lung specimens without virus isolation. Twenty nine lung specimens collected from postweaning pigs were isolated for PRRSV. When the PCR products from the 29 lung specimens were digested by the restriction enzymes MluI, HincII, SacII and HaeIII, the RFLP patterns from the 29 lung specimens matched with those from the corresponding PRRSV isolates from each pig. The results suggest that the PCR-based RFLP analysis method may be useful to distinguish PRRSV isolates directly from lung specimens without virus isolation. PMID- 11411507 TI - Recent epidemiological status of canine viral enteric infections and Giardia infection in Japan. AB - Epidemiology of canine enteric infections was studied. Rectal swabs collected from 95 dogs presented at animal hospitals during a period from January to June of 2000 were examined for enteric pathogens, including viruses and Giardia lamblia (G. lamblia). Most frequently detected in both diarrheal and normal feces were canine coronavirus (55.4%) and G. lamblia (48.2%). Canine parvovirus type 2 (CPV-2) was specifically associated with diarrheal cases and CPV-2b was the predominant antigenic type. Although canine rotavirus, canine adenovirus, and canine distemper virus were also detected in a small number of diarrheal cases, no evidence for calicivirus infection was obtained. PMID- 11411508 TI - Births of freemartins derived from embryos reconstructed with ear fibroblasts. AB - Although the combination of artificial insemination (AI) and embryo transfer (ET) is effective for preventing large offspring syndrome in clone cattle production, it may cause freemartinism. In this study, 51 reconstructed embryos were transferred to artificially inseminated recipients. Of those 9 twin pregnancies, three delivered male and female offsprings. The females had tufts of long coarse hair and short blind pouch at the vaginal end. At necropsy, hypoplastic testicles and epididymis, which connected to the uterus through the spermatic cord, were found and seminal vesicles were also noted. All females had mixed sex chromosome configuration (60, XX and 60, XY). These results suggest that the combined ET program can cause freemartinism, which reduces the efficiency of clone cattle production. PMID- 11411509 TI - Relationship between testicular transferrin and plasma estradiol-17beta concentrations of dogs with azoospermia and dogs with sertoli cell tumors. AB - Testicular Transferrin (Tf) and peripheral plasma estradiol-17beta (E2) concentrations were measured in 3 dogs with azoospermia (AZ dogs), 3 dogs with Sertoli cell tumors (SC dogs), and 5 normal male Beagles. The mean Tf concentrations in the testes of the AZ dogs and the affected testes of the SC dogs, and the plasma E2 concentrations in both these groups of dogs were significantly higher than the values in normal dogs (P<0.05, 0.01 and 0.01, respectively). Therefore, excessive E2 secretion by hyperfunctioning Sertoli cells is thought to have caused the azoospermia in the 3 dogs. PMID- 11411510 TI - Hypertrophic cardiomyopathy in two kittens. AB - Two 2-month-old kittens presented with a loud cardiac murmur. One cat showed severe signs of heart failure such as respiratory effort and exercise intolerance. Echocardiography revealed left ventricular concentric hypertrophy and severe left ventricular outflow obstruction. They died at 5 and 12 months of age, respectively. Necropsy and histopathology confirmed hypertrophic cardiomyopathy. PMID- 11411511 TI - Iron content of rat serum ferritin. AB - The serum ferritin concentration was significantly higher in female than in male rats, reflecting higher iron stores in females than in males. The mean iron/protein ratio of serum ferritin was 0.018+/-0.008 (SD) (microg of Fe/microg of protein) in female rats and 0.011+/-0.011 in male rats, being much lower than that of liver ferritin (0.233+/-0.014 in females and 0.227+/-0.020 in males). Iron loading of rats significantly increased serum ferritin concentration, but did not influence the iron content of serum ferritin. These results indicate that rat serum ferritin contains only a small amount of iron independent of body iron stores. PMID- 11411512 TI - An experimental approach to study the binding properties of vitamin E (alpha tocopherol) during hairless mouse skin permeation. AB - An experimental approach to study the binding properties of vitamin E has been developed. Total vitamin E solubility in the skin was determined by a partition study, followed by in vitro skin permeation studies with whole skin and stripped skin. The amount of freely diffusable vitamin E in the diffusion process was determined from the permeation profiles of whole skin and stripped skin by employing a bi-layer model. The concentrations of vitamin E in the stratum corneum and viable dermis were determined separately. By subtracting this amount from the total concentration of vitamin E in the skin, as determined by the solubility study, the amount of bound vitamin E was determined. After skin permeation reached a steady state, the donor solution was removed and the permeation study continued (desorption study). During the entire period of the desorption experiment, the amount of vitamin E in the receptor solution hardly increased and remained constant. After the desorption experiment, vitamin E still remaining in the skin was determined by extracting with tissue solubilizer, SOLABLE, and is considered as the amount of vitamin E strongly bound in the skin. The concentrations of bound vitamin E determined by permeation and desorption studies coincided relatively well. To further investigate skin binding of vitamin E, a differential scanning calorimetry study was performed. Vitamin E-treated stratum corneum showed phase transitions at 76 and 85 degrees C, associated with lipid transitions. The thermal transitions associated with the lipid transition suggested interactions of vitamin E with lipid components of the skin. During skin permeation, vitamin E forms a very strong reservoir in the skin tissue and this amount of vitamin E, about 30%, exists as a bound-form with the lipid components of the stratum corneum. PMID- 11411513 TI - Selective flotation-spectrophotometric determination of trace copper(II) in natural waters, human blood and drug samples using phenanthraquinone monophenylthiosemicarbazone. AB - Copper(II) forms 1:1 and 1:2 intense red complexes with phenanthraquinone monophenylthiosemicarbazone (PPT) at pH 3-3.5 and > or =6.5, respectively. These complexes exhibit maximal absorbance at 545 and 517 nm, the molar absorptivity being 2.3 x 10(4) and 4.8 x 10(4) l mol(-1) cm(-1), respectively. However, the 1:1 complex was quantitatively floated with oleic acid (HOL) surfactant in the pH range 4.5-5.5, providing a highly selective and sensitive procedure for the spectrophotometric determination of CuII. The molar absorptivity of the floated Cu-PPT complex was 1.5 x 10(5) l mol)(-1) cm(-1). Beer's law was obeyed over the range 3-400 ppb at 545 nm. The analytical parameters affecting the flotation process and hence the determination of copper traces were reported. Also, the structure of the isolated solid complex and the mechanism of flotation were suggested. Moreover, the procedure was successfully applied to the analysis of CuII in natural waters, serum blood and some drug samples. PMID- 11411514 TI - Mechanistic appraisal of the charge-transfer complexes of promethazine with chloranil: a modelling approach. AB - Various mechanisms are often used to explain the interaction between electron donors and acceptors. Commonly proposed mechanisms are those in which the acceptor interacts with the aromatic pi-systems in the donor molecule or the acceptor forms a weak interaction of the Lewis acid with Lewis base type. In this study, the above mechanisms were examined as well as other possible mechanisms. Promethazine was chosen as the model drug containing aromatic systems capable of pi-pi interaction as well as N-methyl group capable of forming a complex with the weak Lewis acid, p-chloranil. Our modelling studies revealed that the situation where the p-chloranil interacts with a protonated N-methyl group is the most significant mechanism of interaction, based on the calculated energies for the highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO), the Tripos force field energy terms and also the stability of the complexes during molecular dynamics simulations. PMID- 11411515 TI - Synthesis and cytotoxic activity of benzopyranoxanthone analogues of benz. AB - Condensation of 3-hydroxy-2-naphthalenecarboxylic acid with phloroglucinol afforded 1,3-dihydroxy-12H-benzo[b]xanthen-12-one. Construction of an additional dimethylpyran ring onto this skeleton, by alkylation with 3-chloro-3-methyl-1 butyne followed by Claisen rearrangement, gave access to a series of benzo[b]pyrano[2,3-i]xanthen-6-ones and benzo[b]pyrano[3,2-h]xanthen-7-ones related to psorospermine and benzo[b]acronycine. In contrast with what is observed in the pyridoacridone and benzopyridoacridone series, the linear benzo[b]-pyrano[2,3-i]xanthen-6-one derivatives were more potent than their angular benzo[b]pyrano[3,2-h]xanthen-7-one isomers. cis-3,4-Diacetoxy-5-methoxy 2,2-dimethyl-3,4-dihydro-2H,6H-benzo[b]pyrano[2,3-i]xanthen-6-one, the most active among the new compounds, was more potent than acronycine in inhibiting the proliferation of L1210 murine leukemia cells. PMID- 11411516 TI - Prediction of endocrine disruptors based on a new structure-activity relationship for sex and environmental hormones using chemical hardness concept. AB - Classification of the relationship between electronic structures and biological activities of endocrine disruptors (so-called environmental hormones) was attempted using the parameters of absolute hardness (eta), absolute electronegativity (chi), and global softness (S), approximately defined as eta=1/2(epsilonLUMO-epsilonHOMO), chi=-1/2(epsilonHOMO+ epsilonLUMO), and S=1/eta, respectively, based on the hardness concept. The strength of binding affinity and toxicity of the chemicals were approximately proportional to the absolute hardness, and laterally toxic chlorinated PCDDs, PCBs, and DDTs are classified as chemically soft. Here we found that the electronic structures of environmental hormones can be classified into four main groups: 17beta-estradiol type (group I), testosterone type (group II), thyroxine type (group III), and HCH (hexachlorocyclohexane) type (group IV). Therefore, if we can predict the coordinate (chi, eta) of the electronic structure of one chemical on the eta-chi activity diagram, we would be able to predict the receptor with which the chemicals (environmental hormones) interact. For instance, 2,3,7,8 tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) is classified in group II, therefore, it would bind with the thyroid receptor more than the estrogen receptor (group I). It appears that dibutyl phthalate would not interact with estrogen receptor because it does not belong to group I. In addition, the coordinates of these four groups do not complementarily overlap with the electronic structures of 20 natural amino acid residues. The eta-chi activity diagram is a new tool for the prediction of the toxicity and biological activity of environmental hormones. PMID- 11411517 TI - Novel products from oxidation of the norditerpenoid alkaloid pseudaconine with HIO4. AB - Oxidation of pseudaconine 8, a norditerpenoid alkaloid, with HIO4 led to a series of novel interesting products, depending greatly on reaction medium and work-up conditions. Treatment of 8 in MeOH-H2O (1:1) with NaIO4 gave compounds 10 and 11, but compound 12 was obtained quantitatively when the final reaction solution was alkalized with conc. NH4OH. The imine 12 was also obtained in 100% yield by treating 8 in 5% HCl solution with NaIO4 followed by alkalizing the reaction products to pH>9 with conc. NH4OH. When the work up pH was 7-8, only N,O-mixed acetal-ketal 13 was formed in 96% yield, which was converted quantitatively to 12 by further alkalizing. When the reaction mixture was alkalized to pH 7-8 with Na2CO3, a hemiacetalketal 14 was afforded quantitatively, which was converted to 15 in 87% yield by further treatment with Na2CO3 or 5% NaOH methanol. Compound 15 could be converted back to 14 by treatment with 10% HCl solution. Acetylation of the imine 12 gave the compounds 16 and 17 in 15% and 19% yields, respectively. All of the new compounds were isolated and fully characterized. PMID- 11411518 TI - Constituents of crinoidea. 2. Isolation and structure of the novel type gangliosides from the feather star Comanthus japonica. AB - Two novel type gangliosides CJP2 and CJP3 have been obtained from the feather star Comanthus japonica. On the basis of methylation linkage analysis combined with ammonolysis and other chemical and spectroscopic evidence, the chemical structures of CJP2 and CJP3 were determined to be alpha-9-O-Me-NeuGc-(2-->3) inositolphosphoceramide and alpha-9-O-Me-NeuGc-(2-->11)-alpha-9-O-Me-NeuGc-(2- >3)-inositolphosphoceramide, respectively. These gangliosides are unique in that they are inositolphosphoceramide derivatives possessing sialic acid; such gangliosides have not previously been identified. The presence of 9-O-methyl-N glycolyl-neuraminosyl residues is also unique in naturally occurring gangliosides. PMID- 11411519 TI - An iridoid gentiobioside, a benzophenone glucoside and acylated flavone C glycosides from Tripterospermum japonicum (Sieb. et Zucc.) Maxim. AB - From aerial parts of Tripterospermum japonicum, 6'-O-beta-D glucopyranosylmorroniside, benzophenone glucoside, named triptephenoside and 2''' and 4'''-O-acetyl-2''-O-alpha-L-rhamnopyranosylisovitexins were isolated, along with known iridoid and secoiridoid glucosides, and C-glycosyl flavones. PMID- 11411520 TI - Synthesis and azannulation of pyridinylaminohexadienones. AB - 4-(2-Pyridinylamino)-1,1,1-trifluoro-3-penten-2-ones 3, obtained from the reaction of commercially available 2-aminopyridine derivatives and 4-methoxy 1,1,1-trifluoro-3-penten-2-one 2, were converted to 6-(dimethylamino)-4-(2 pyridinylamino)-3,5-hexadien-2-ones 4 by treatment with dimethylformamide dimethyl acetal. Azannulation of hexadienones 4 afforded 4-(2-pyridinylamino)-2 trifluoromethylpyridines 5 and 2-(trifluoroacetylmethylene)pyrido[1,2 a]pyrimidines 6, classes of compounds particularly interesting from a chemical and biological point of view. PMID- 11411521 TI - Farnesyl hydroxybenzoic acid derivatives from Ferula kuhistanica. AB - Four new farnesyl hydroxybenzoic acid derivatives, kuhistanols E-H (1-4), were isolated from the roots of the Uzbekistan medicinal plant Ferula kuhistanica. The structures of the new compounds were elucidated based on spectroscopic and chemical evidence. PMID- 11411522 TI - Studies on chemical modification of monensin IX. Synthesis of 26-substituted monensins and their Na+ ion transport activity. AB - The C-26 modified monensin derivatives, 26-O-benzoylmonensin (3), 26-O benzylmonensin (4) and 26-phenylaminomonensin (5) were prepared from monensin (1). Na+ ion transport activity through biological membrane and antibacterial activity of 3-5 were evaluated and compared with the activities reported for a 26 phenylurethane derivative (2). Among these compounds, 5 showed the largest Na+ ion transport and antibacterial activities. In these compounds, the formation of head-to-tail hydrogen bonds was suggested to be an important factor for Na+ ion transport and antibacterial activities. PMID- 11411523 TI - Effects of constituents from the bark of Magnolia obovata on nitric oxide production in lipopolysaccharide-activated macrophages. AB - The methanolic extract from a Japanese herbal medicine, the bark of Magnolia obovata, was found to inhibit nitric oxide (NO) production in lipopolysaccharide (LPS)-activated macrophages. By bioassay-guided separation, three neolignans (magnolol, honokiol, obovatol) and three sesquiterpenes (alpha-eudesmol, beta eudesmol, gamma-eudesmol) were obtained as active constituents. A trineolignan (magnolianin), a phenylpropanoid glycoside (syringin), lignan glycosides (liriodendrin, (+)-syringaresinol 4'-O-beta-D-glucopyranoside) and a sesquiterpene (caryophyllene oxide) did not show any activity. On the other hand, sesquiterpene-neolignans (eudesmagnolol, clovanemagnolol, caryolanemagnolol, eudeshonokiol A, eudesobovatol A) showed the strong cytotoxic effects. Active constituents (magnolol, honokiol, obovatol) showed weak inhibition for inducible NO synthase (iNOS) enzyme activity, but potent inhibition of iNOS induction and activation of nuclear factor-kappaB. PMID- 11411524 TI - Synthesis and antimalarial activity of febrifugine derivatives. AB - The regioisomers (2a,b) of the piperidine ring of febrifugine (1a) and isofebrifugine (1b) were synthesized from 4-allyl-3-piperidone (5). Reduction of 5 afforded a mixture of the trans and cis alcohols (6a,b) without diastereoselectivity; this result differentiated it from the reduction of 2-allyl 3-piperidone (14). The antimalarial activity of 2a,b and related compounds was tested. PMID- 11411525 TI - Steroidal glycosides from the bulbs of Camassia leichtlinii and their cytotoxic activities. AB - Phytochemical analysis of the bulbs of Camassia leichtlinii (Liliaceae) resulted in the isolation of six new spirostanol saponins, a new furostanol saponin, a cholestane glucoside, and four known steroidal saponins. The structures of the new saponins were determined by detailed analysis of their spectral data, including two-dimensional NMR spectroscopy, and by the results of hydrolytic cleavage. Cytotoxic activities of the isolated compounds against human oral squamous cell carcinoma (HSC-2) cells and normal human gingival fibroblasts (HGF) are also reported. PMID- 11411526 TI - Studies on the constituents of Catalpa species. VI. Monoterpene glycosides from the fallen leaves of Catalpa ovata G. Don. AB - Five new monoterpene glycosides, ovatolactone 7-O-(6'-O-p-hydroxybenzoyl)-beta-D glucopyranoside, ovatic acid methyl ester 7-O-(6'-O-p-hydroxybenzoyl)-beta-D glucopyranoside, 7-O-p-hydroxybenzoylovatol 1-O-(6'-O-p-hydroxybenzoyl)-beta-D glucopyranoside, 6'-O-p-hydroxybenzoylcatalposide and (2E,6R)-2,6-dimethyl-8 hydroxy-2-octenoic acid 8-O-[6'-O-(E)-p-coumaroyl]-beta-D-glucopyranoside were isolated from the fallen leaves of Catalpa ovata G. Don. Their structures were determined by extensive spectroscopic studies and syntheses. PMID- 11411527 TI - Alkaloids, diarylheptanoid and naphthalene carboxylic acid ester from Rhoiptelea chiliantha. AB - Two pyrrolidine alkaloids (1, 2) were isolated from the fruits of Rhoiptelea chiliantha Diel et Hand.-Mazz. (Rhoipteleaceae). A diphenyl ether-type diarylheptanoid (3), and a naphthalene carboxylic acid methyl ester (4) which is biogenetically-related to juglone were isolated from the branches of the same plant. Their chemical structures were elucidated on the basis of spectroscopic analysis and chemical evidence. PMID- 11411528 TI - Bioactive saponins and glycosides. XVIII. Nortriterpene and triterpene oligoglycosides from the fresh leaves of Euptelea polyandra Sieb. et Zucc. (2): Structures of eupteleasaponins VI, VI acetate, VII, VIII, IX, X, XI, and XII. AB - Following the elucidation of eupteleasaponins I, II, III, IV, V, and V acetate, eupteleasaponins VI, VI acetate, VII, VIII, IX, X, XI, and XII were isolated from the fresh leaves of Euptelea polyandra Sieb. et Zucc. The structures of eupteleasaponins VI-XII were determined on the basis of chemical and physicochemical evidence. PMID- 11411529 TI - Relationship of electrochemical oxidation of catechins on their antioxidant activity in microsomal lipid peroxidation. AB - The oxidation potentials of catechins were measured by employing flow-through column electrolysis. The oxidation potentials of catechins were shown to depend on their structures. At the same time, the antioxidant activity of catechins on NADPH-dependent lipid peroxidation in rat liver microsomes was evaluated. Catechins showed a 50% inhibition of lipid peroxidation in the concentration range of 10-51 microM. Among those studied, galloylated catechins exhibited stronger antioxidant activities than those of nongalloylated catechins. A quantitative relationship has been obtained to describe the antioxidant activity of catechins: log IC50 (microM)= 1.56+2.49E1/2 (V)-0.29 logP (r=0.907), where IC50 represents the concentration for 50% inhibition of lipid peroxidation, E1/2 represents the half-wave potential of the first oxidation wave, and P represents the octanol/water partition coefficient. This relationship suggested two important characteristics determining catechin antioxidant activity, namely the ease of oxidation and the lipophilicity. PMID- 11411530 TI - Triterpenoid saponins from berries of Hedera colchica. AB - Fifteen triterpenoid saponins were isolated from the berries of Hedera colchica and their structures established on the basis of chemical and spectroscopic evidence. Among them, two are new compounds: coichiside A (3) and colchiside B (15) and four are described for the first time in the berries of Hedera colchica (compounds 1, 8, 9 and 11). PMID- 11411531 TI - Glucose and lactate biosensors prepared by a layer-by-layer deposition of concanavalin A and mannose-labeled enzymes: electrochemical response in the presence of electron mediators. AB - An electrochemical response of glucose and lactate biosensors which were prepared by coating a platinum electrode with a thin film composed of concanavalin A and mannose-labeled glucose oxidase (GOx) or lactate oxidase (LOx) was evaluated in the presence of ferrocene derivatives as electron mediator. Both glucose and lactate biosensors showed catalytic current to glucose and lactate, respectively, in cyclic voltammetry, suggesting that the ferrocene derivatives can mediate electron transport smoothly from the reduced forms of GOx and LOx in the thin films to the electrode. Among the three kinds of ferrocene derivatives used, ferrocenylmethanol was found to be the most suitable electron mediator because of its low oxidation potential. The glucose and lactate sensors gave useful calibration graphs, in which higher detection limits were reached as compared with those observed when the sensors were operated in the absence of electron mediator. PMID- 11411532 TI - New triterpenoid saponins from Maesa tenera. AB - Two new triterpenoid saponins, maetenosides A and B were isolated from the aerial parts of Maesa tenera and were respectively defined as 3-O-[beta-D-glucopyranosyl (1-->2)-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-galactopyranosyl-(1-->3)][beta-D glucopyranosyl-(1-->2)]-beta-D-glucuronopyranosyl camelliagenin A 22-O-angelate (1) and 3-O-[alpha-L-rhamnopyranosyl-(1-->2)-beta-D-galactopyranosyl-(1- >3)][beta-o-glucopyranosyl-(1-->2)]-beta-D-glucuronopyranosyl camelliagenin A 22 O-angelate (2). Their structures were established on the basis of chemical and spectroscopic methods. PMID- 11411533 TI - Purification and characterization of anti-complementary polysaccharide from leaves of Thymus vulgaris L. AB - For the purification of the anti-complementary polysaccharide from Thymus vulgaris L., the hot-water extract of thyme leaves was successively fractionated by ethanol precipitation and ultra-filtration. The retentate with 300-kDa membrane cartridge showed a potent anti-complementary activity. It was further purified by open column chromatographies on DEAE-Toyopearl 650C, Butyl-Toyopearl 650M and Sephadex G-100, obtaining TV3-IIA-I, the purified anti-complementary polysaccharide. The anti-complementary polysaccharide exhibited the anti complementary activity via both classical and alternative pathways. Based upon the methylation analysis and the reaction with beta-glucosyl Yariv reagent, the anti-complementary polysaccharide from thyme might contain an arabinogalactan moiety, at least in part. PMID- 11411534 TI - Latifolosides K and L, two new triterpenoid saponins from the bark of Llex latifolia. AB - Two new triterpenoid saponins, latifoloside K (1), 3-O-beta-D-glucopyranosyl-(1- >3)-[alpha-L-rhamnopyranosyl-(1-->2)-]-alpha-L-arabinopyranosyl 3beta-hydroxy-urs 12,18-dien-28-oic acid 28-O-beta-D-glucopyranosyl ester and latifoloside L (2), 3 O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->3)-[alpha-L rhamnopyranosyl-(1-->2)-]-alpha-L-arabinopyranosyl 3beta,19alpha-dihydroxyursolic acid, were isolated from the bark of Ilex latifolia Thunb. Also isolated were two known compounds, ilekudinoside A (3) and kudinoside G (4). Structural assignments were established on the basis of spectroscopic data and chemical evidence. PMID- 11411535 TI - Three new flavonol malonylrhamnosides from Ribes alpinum. AB - Three new flavonol malonylrhamnosides, 3-O-(4"-O-malonyl)-alpha-L rhamnopyranosides of mearnsetin, myricetin and quercetin respectively, together with the corresponding mearnsitrin, myricitrin, quercitrin and the 4-O-methyl phloracetophenone 2-O-beta-D-glucopyranoside, were isolated from the leaves of Ribes alpinum and fully characterized by spectrocopic methods including 2D NMR. PMID- 11411536 TI - A new secoiridoid glucoside, amaronitidin, from the Peruvian folk medicine "Hercampuri" (Gentianella nitida). AB - A new secoiridoid glucoside designated amaronitidin (1) was isolated from the Peruvian folk medicine "Hercampuri" (Gentianella nitida) along with three known secoiridoid glucosides. Their structures were determined by extensive spectroscopic investigation. PMID- 11411537 TI - Two new podophyllotoxin glucosides from Sinopodophyllum emodi (Wall.) Ying. AB - Two new aryltetralin-type lignans, isopodophyllotoxin 7'-O-beta-D-glucopyranosyl (1-->6)-beta-D-glucopyranoside (1) and 4-demethyl-picropodophyllotoxin 7'-O-beta D-glucopyranoside (2), along with eight known podophyllotoxin derivatives: 4 demethyl-podophyllotoxin 7'-O-beta-D-glucopyranoside (3), podophyllotoxin 7'-O beta-D-glucopyranoside (4), deoxypodophyllotoxin (5), picropodophyllotoxin (6), podophyllotoxin (7), 4-demethyl-picropodophyllotoxin (8), 4-demethyl podophyllotoxin (9), and 4-demethyl-deoxypodophyllotoxin (10), were isolated from the roots and rhizomes of Sinopodophyllum emodi (Wall.) Ying (Berberidaceae). Their structures were identified based on NMR spectral data and chemical evidence. PMID- 11411538 TI - Medicinal foodstuffs. XXIII. Structures of new oleanane-type triterpene oligoglycosides, basellasaponins A, B, C, and D, from the fresh aerial parts of Basella rubra L. AB - Basellasaponins A, B, C, and D, oleanane-type triterpene oligoglycosides having the dioxolane-type substituent, were isolated from the fresh aerial parts of Basella rubra L. together with betavulgaroside I, spinacoside C, and momordins IIb and IIc. The chemical structures of basellasaponins A, B, C, and D were determined from chemical and physicochemical evidence. PMID- 11411539 TI - Antioxidative glucosides from the fruits of Ligustrum lucidum. AB - The ethanol extract of the fruits of Ligustrum lucidum was shown to have inhibitory effects on the hemolysis of red blood cells induced by 2,2'-azo-bis-(2 amidinopropane) dihydrochloride. Bioassay-guided analysis led to the isolation of ten secoiridoid glucosides. Two of them were new, lucidumosides C and D. Their structures were elucidated by spectroscopic methods. The other eight compounds were identified as oleoside dimethyl ester, ligustroside, oleuropein, nuezhenide, isonuezhenide, neonuezhenide, lucidumoside A and lucidumoside B. Five compounds, oleoside dimethyl ester, oleuropein, neonuezhenide, lucidumoside B and lucidumoside C, exhibited strong antioxidant effect against hemolysis of red blood cells induced by free radicals. PMID- 11411540 TI - Hopeafuran and a C-glucosyl resveratrol isolated from stem wood of Hopea utilis. AB - A new resveratrol dimer and a new C-glucosyl resveratrol were isolated from stem wood of Hopea utilis along with nine stilbenoid derivatives comprising bergenin and (+)-lyoniresinol. The structures have been elucidated on the basis of the spectroscopic evidence. PMID- 11411541 TI - An improved synthesis of butyl 4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-1 piperidineacetate (AU-224). AB - A new and facile route for the synthesis of the novel gastrointestinal prokinetic butyl 4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-1-piperidineacetate (1b), which exhibited potent gastro- and colon-prokinetic activities by oral administration without significant side effects, was established. The key intermediate, butyl 4-amino-1-piperidineacetate (16), was prepared from commercially available 4-amino-1-benzylpiperidine (2) in a high yield with four steps. Compound 1b was prepared by condensation of commercially available 4-amino 5-choloro-2-methoxybenzoic acid (7) with 16 in 84% yield. This improved synthetic route was appropriate for large-scale synthesis of 1b. PMID- 11411542 TI - Alpha-glucosidase inhibitors with a 4,5,6,7-tetrachlorophthalimide skeleton pendanted with a cycloalkyl or dicarba-closo-dodecaborane group. AB - Previous studies of alpha-glucosidase inhibitors derived from thalidomide revealed that 4,5,6,7-tetrachloro-N-alkylphthalimide derivatives are superior lead compounds. Structure-activity relationship studies indicated that a hydrophobic group at the N(2) position is mandatory for potent activity. Accordingly, we have designed and synthesized some 4,5,6,7-tetrachloro-N cycloalkylphthalimide and 4,5,6,7-tetrachloro-N-dicarba-closo-dodecaborane derivatives. The prepared compounds exhibited potent alpha-glucosidase-inhibitory activity. Among them, 4,5,6,7-tetrachloro-N-cycloheptylphthalimide (9) showed the most potent activity, being approximately 30 times more active than the classical inhibitor, 1-deoxynojirimycin (1). PMID- 11411543 TI - Significant expression of functional human type 1 mitochondrial ADP/ATP carrier in yeast mitochondria. AB - As a first step to characterize the unknown functional properties of the human mitochondrial ADP/ATP carrier (AAC), we tried to express human type 1 AAC (hAAC1) in Saccharomyces cerevisiae. Expression of hAAC1 in yeast mitochondrial membrane was very low, although its transcript level was high. Its expression was improved greatly by replacement of its N-terminal region with the corresponding region of yeast type 2 AAC (yAAC2), as observed with the bovine type 1 AAC (bAAC1). This chimeric hAAC1 showed similar ADP transport activity to that of chimeric bAAC1, corresponding to the transport activity of bAAC1 in bovine heart mitochondria. These results suggested that the N-terminal region of yAAC2 is important for expression of the mammalian carriers in yeast mitochondria. Using the present expression system, studies on the functional properties of the human AAC isoforms in relation to their structures are now possible. PMID- 11411544 TI - Accumulation of medium chain acyl-CoAs during beta-oxidation of long chain fatty acid by isolated peroxisomes from rat liver. AB - We have reported fatty alcohol synthesis accompanied by chain elongation in liver peroxisomes (Biochim. Biophys. Acta, 1346, 38 (1997)). In the present experiment, we studied what kind of acyl-CoA(s) destined to be utilized as primer for fatty alcohol synthesis accumulate(s) during peroxisomal beta-oxidation. Peroxisomes were prepared from rat liver treated with clofibrate, a peroxisome proliferator, and incubated with [U-14C]palmitate, in order to investigate acyl-CoAs after beta oxidation. At 1 mM concentration, MgATP activated beta-oxidation, but inhibited beta-oxidation at concentrations higher than 1 mM. After incubation of peroxisomes with palmitate, various acyl-CoAs were formed. Among medium-chain labelled acyl-CoAs, octanoyl-CoA was mainly detected. These results suggest that octanoyl-CoA accumulates during beta-oxidation of palmitate. When peroxisomes were incubated with [9,10-(3)H]palmitate and [9,10-(3)H]stearate, among medium chain acyl-CoAs, octanoyl-CoA and decanoyl-CoA were primarily detected, respectively, suggesting the occurrence of at least 4 cycles of beta-oxidation of both fatty acids by peroxisomes. PMID- 11411545 TI - Biological activity of alpha-thujaplicin, the minor component of Thujopsis dolabrata SIEB. et ZUCC. var. hondai MAKINO. AB - Alpha-thujaplicin, a minor component of Thujopsis dolabrata SIEB. et ZUCC. var. hondai MAKINO, which was synthesized, showed the antibacterial activity, phytogrowth-inhibitory effect, inhibition of carboxypeptidase A and cytotoxic effect. Antibacterial activity of alpha-thujaplicin on Enterococcus faecalis IFO 12965 [minimum inhibitory concentration (MIC): 1.56 microg/ml] was higher than that of gentamicin (MIC: 6.25 microg/ml) used as a positive control. Inhibitory activity of alpha-thujaplicin on carboxypeptidase A [50% inhibitory concentration (IC50): 3.24 x 10(-5) M] was higher than that of 1,10-phenanthroline used as a positive control. Alpha-thujaplicin showed germination inhibition toward the seed of Echinochloa utilis Ohwi et Yabuno even at the low concentration of 10 ppm and its growth inhibitory effect was stronger than that of sodium 2,4 dichlorophenoxyacetate used as a standard. Alpha-thujaplicin at 1.25 microg/ml inhibited cell growth of human stomach cancer KATO-IIl by 86%, and Ehrlich's ascites carcinoma by 87%, respectively. This compound even at the low concentration of 0.32 microg/ml also inhibited cell growth of the former by 66%, and the latter by 75%, respectively. The acute toxicity of alpha-thujaplicin [50% lethal dose (LD50) value: 256 mg/kg] in mice was as strong as those of beta dolabrin (LD50 value: 232 mg/kg) and gamma-thujaplicin (LD50 value: 277 mg/kg). PMID- 11411546 TI - Reversal of vinblastine resistance in human leukemic cells by haloperidol and dihydrohaloperidol. AB - Haloperidol, an antipsychotic, was investigated in cells overexpressing P glycoprotein to detemine whether it was a clinically effective drug to reverse for reversing multidrug resistance (MDR) mediated by P-glycoprotein. A nontoxic concentration of haloperidol (1-30 microM) enhanced the cytotoxic effects of vinblastine (VBL) concentration-dependently in VBL-resistant human leukemia (K562/VBL) cells, but had no effect in the parent cells. Haloperidol also enhanced the cytotoxicities of epirubicin, doxorubicin and actinomycin D in the K562/VBL cells, but not those of idarubicin or cisplatin; this enhancement was less than that of the VBL toxicity in the VBL-resistant tumor line. Haloperidol increased the intracellular accumulation of VBL in the K562/VBL cells, and the binding of [3H]-azidopine to the cell-surface protein, P-glycoprotein, was inhibited by haloperidol in a concentration-dependent manner. Haloperidol was less potent than verapamil. Thus, haloperidol appeared to potentiate anticancer agents through the reversal of MDR by competitively inhibiting drug-binding to P glycoprotein. In contrast, the main metabolite of haloperidol, dihydrohaloperidol, without antipsychotic activity, had less of an effect. Therefore, haloperidol might be useful in reversing drug-resistance. PMID- 11411547 TI - Antiplatelet effect of 2-chloro-3-(4-acetophenyl)-amino-1,4-naphthoquinone (NQ301): a possible mechanism through inhibition of intracellular Ca2+ mobilization. AB - The effects of 2-chloro-3-(4-acetophenyl)-amino-1,4-naphthoquinone (NQ301), an antithrombotic agent, on aggregation, binding of fibrinogen to glycoprotein (GP)IIb/IIIa complex and intracellular signals were investigated using human platelets. NQ301 significantly inhibited the collagen-, thrombin-, arachidonic acid-, thapsigargin- and calcium ionophore A23187-induced aggregation of washed human platelets with IC50 values of 13.0+/-0.1, 11.2+/-0.5, 21.0+/-0.9, 3.8+/-0.1 and 46.2+/-0.8 microM, respectively. NQ301 also significantly inhibited FITC conjugated fibrinogen binding to human platelet surface GPIIb/IIIa complex, but failed to inhibit the fibrinogen binding to purified GPIIb/IIIa complex. These data demonstrate that NQ301 inhibits platelet aggregation by suppression of the intracellular pathway, rather than by direct inhibition of fibrinogen-GPIIb/IIIa complex binding. NQ301 significantly inhibited the increase of cytosolic Ca2+ concentration and ATP secretion, and also significantly increased platelet cAMP levels in the activated platelets. These results suggest that the antiplatelet activity of NQ301 may be mediated by inhibition of cytosolic Ca2+ mobilization, enhancement of cAMP production and inhibition of ATP secretion in activated platelets. PMID- 11411548 TI - In vitro effects of wood creosote on enterotoxin-induced secretion measured electrophysiologically in the rat jejunum and colon. AB - Secretory diarrhea occurs when the balance between intestinal absorption and secretion is disturbed by excessive secretion caused by enterotoxins produced by the pathogen. Wood creosote has long been used as a traditional antidiarrheal remedy. The goal of our study was to extend our knowledge about the antisecretory action of wood creosote against Escherichia coli enterotoxin-induced secretion in the small intestine and colon. Experiments were performed in mucosal sheets of rat jejunum and colon which were stripped of the external muscle layers to eliminate interactions with smooth muscle activity and local blood flow. Mucosal sheets were placed in modified Ussing chambers and hypersecretory conditions were induced by heat-labile (LT) or heat-stable (STa) E. coli enterotoxins added cumulatively (0.01-10 microg/ml) to the mucosal bathing solution. Intestinal secretion was monitored electrophysiologically as transmucosal short circuit current (Isc). LT induced a concentration-dependent increase in Isc in the rat jejunum, with no effect in the colon. In contrast, STa induced a significant increase in colonic Isc, without causing any change in Isc across the jejunum. In separate experiments the effects of increasing concentrations of wood creosote (0.1-50 microg/ml), added to the mucosal or serosal bathing solution, were examined against the secretory responses induced by LT or STa. In the small intestine the antisecretory activity of wood creosote against LT-induced secretion was more potent following serosal application, whereas in the colon wood creosote inhibited STa-induced secretion with equal potency following either serosal or mucosal addition. In summary, our findings demonstrate that wood creosote possesses antidiarrheal activity suppressing E. coli enterotoxin-induced secretion in both the small intestine and colon. PMID- 11411549 TI - Effects of various antihypertensive drugs on the function of osteoblast. AB - Several studies have suggested that high blood pressure is associated with the risk of bone loss. Since various antihypertensive drugs are in wide use for the treatment of hypertension, it is important to investigate the influences of these drugs on bone metabolism. Osteoblasts play a pivotal role in the regulation of bone formation. During differentiation, they sequentially express type I collagen, alkaline phosphatase (ALP), other bone matrix proteins, and finally undergo mineral deposition. In this study, we examined the effects of various antihypertensive drugs on the function of osteoblast using clonal MC3T3-E1 cells. Drugs examined include dihydropyridine-type calcium channel blockers (benidipine, amlodipine, and nifedipine), angiotensin-converting enzyme (ACE) inhibitors (captopril, lisinopril, and enalapril), and angiotensin II receptor type1 (AT1) antagonists (TCV-116 and KW-3433). None of the ACE inhibitors or AT1 antagonists affected ALP activity or cellular DNA content significantly. In contrast, benidipine, amlodipine, and nifedipine increased ALP activity when used in amounts 1 pM, 100 nM, and 100 nM, respectively. Benidipine blocked calcium influx through the L-type voltage dependent calcium channel of MC3T3-E1 more potently than amlodipine or nifedipine. These calcium channel blockers did not change collagen accumulation. Benidipine significantly increased in vitro mineralization at a concentration of 1 nM and higher, while amlodipine did so at 1 microM and nifedipine did not. Comparison of the effective concentration of each calcium channel blocker in our study with the reported maximum serum concentration of each drug suggests that benidipine, but not amlodipine or nifedipine, promotes mineral deposition in human. PMID- 11411550 TI - Effect of glucocorticoid on expression of rat MUC5AC mRNA in rat gastric mucosa in vivo and in vitro. AB - The gastric mucus is an important factor in the gastric mucosal protection from acid, pepsin and various reagents (alcohol, aspirin, etc.). MUC5AC is the mucin secreted from surface mucous cells, and belongs to the gel-forming mucin. We examined the regulation of rat MUC5AC (rMUC5AC) mRNA by glucocorticoid in vivo and in vitro, comparing it with that of pepsinogen (Pg) mRNA. By adrenal gland resection, rMUC5AC and Pg mRNA levels and Pg content in rats significantly decreased to 70%, 46% and 42% of those in the sham operated controls, respectively. With the treatment of hydrocortisone (1, 5 and 50 mg/kg), Pg mRNA level and Pg content in adrenalectomized rats was restored. On the other hand, the rMUC5AC mRNA level exceeded the control with 1 or 5 mg/kg injections of hydrocortisone, but drastically decreased to 18% of sham operation levels with it (50 mg/kg). Similar results were obtained in normal rats with the treatment of hydrocortisone (50 mg/kg). Mucus and DNA content of cultured rat gastric epithelial cells were not affected by hydrocortisone, but rMUC5AC mRNA level was significantly decreased in a dose-dependent manner. From the in vivo and in vitro results, at least a physiological concentration of glucocorticoid was necessary in the expression of rMUC5AC mRNA. However, high dose of hydrocortisone directly suppressed the expression of rMUC5AC mRNA. These results suggested that hydrocortisone might directly cause the suppression and indirectly the enhancement of the mucin biosynthesis. PMID- 11411551 TI - Effects of Shikunshito-Kamiho on fecal enzymes and formation of aberrant crypt foci induced by 1,2-dimethylhydrazine. AB - Shikunshito-Kamiho (SKTK) is a traditional Chinese medicine composed of eight crude drugs (Ginseng Radix, Hoelen, Atractylodis Rhizoma, Glycyrrhizae Radix, Prunellae Spica, Ostreae Testa, Laminaria Thallus, Sargassum). We investigated the effects of SKTK on pH, ammonia, fecal enzymes such as beta-glucuronidase, tryptophanase, urease, and formation aberrant crypt foci in the colon carcinogenesis model induced by 1,2-dimethylhydrazine (DMH). Water extract of SKTK was administered orally for 5 weeks to DMH-treated mice as 0.5% and 1.5% of the diet. Beta-glucuronidase, pH and tryptophanase were significantly inhibited after treatment of 0.5% and 1.5% SKTK, while urease was significantly reduced only during and after treatment of 1.5% SKTK as compared with control data. However, the ammonia concentration wasn't different in SKTK treated groups from control group. The incidence number of aberrant crypts foci (ACF) and aberrant crypts/focus in colon was significantly decreased by 0.5% and 1.5% SKTK mixed diets compared with that in rats treated with DMH alone. These results suggest that SKTK exterts anticarcinogenic activity on experimental murine colorectal cancer. PMID- 11411552 TI - Effect of long term-administration with Gymnema sylvestre R. BR on plasma and liver lipid in rats. AB - Extract of Gymnema sylvestre leaves was administered to rats receiving either a high fat diet or normal fat diet for 10 weeks to investigate its influence on plasma and liver lipids and on visceral fat accumulation. In addition, its effect was compared with those of chitosan and the influence of combined use of these two substances was also evaluated. Within the high fat diet groups, the extract suppressed body weight gain and accumulation of liver lipids to the same extent as chitosan and the combined use. In addition, intraperitoneal fat and fat drop vacuoles on the epithelium of renal tubules, noted in the high fat diet group, were scattered by administration of the extract with the same results as for chitosan and combined use. Within the normal fat diet groups, plasma triglyceride levels decreased by administration of the extract, with similar results as chitosan and combined use. Concerning plasma total cholesterol, there was no decreasing effects with the extract, as found with chitosan and combined use. However, the effect of chitosan on plasma total cholesterol tended to be enhanced when used in combination with the extract. In addition, long-term administration of the extract did not show any influence on hematological and blood chemical parameters. PMID- 11411553 TI - Granulomatous tissue formation of shikon and shikonin by air pouch method. AB - "The extract of shikon" (SK) and shikonin play important roles in the development of granulomatous tissue formation. To reveal the augmenting effect of SK or shikonin on vascular endothelial growth factor (VEGF) production and neovascularization, we investigated murine granulomatous tissue induced by SK and shikonin, comparing them to pouches in which trehalose 6,6'-dimycolate (TDM) was injected. The development of granulomatous tissue formation was evaluated by the wet weight of pouch walls. At day 5 and 7 after SK and shikonin injection, prominent granulomatous tissue formation was detected. Histological observations on the development of granulomatous tissue showed that the pouch was formed in the submuscular connective tissue and necrotic tissue directly facing the cavity and granulomatous tissue developed in the connective tissue. At day 1, VEGF positive neutrophils accumulated in the pouch wall. Granulomatous tissue formation and neovascularization by injection of SK or shikonin was not more prominent than TDM. However, the present results indicate that SK and shikonin induce neovascularization in granulomatous tissue. PMID- 11411554 TI - Oral delivery of synthetic eel calcitonin, elcatonin, in rats. AB - This study was designed to develop an oral dosage form of elcatonin (EC), a hypocalcemic peptide. The EC absorption was estimated by the reduction in plasma calcium concentrations. When EC was orally coadministered with nitroso-N-acetyl D,L-penicillamine (SNAP, 4.0 mg) and 0.02% Carbopol solution or with taurocholate (20 mM) and 0.02% Carbopol solution, the lowering effect was increased compared with that after EC alone, but the F values (0.32 and 0.30%) were extremely small. The oral administration of the mucoadhesive emulsion, which was prepared by coating the W/O/W emulsion with 0.1% Carbopol, enhanced the calcium lowering effect, with the F value of 0.43%. The strong mucoadhesion of the mucoadhesive emulsion to the gastrointestinal mucosa was observed. A capsule containing EC (500 microg), taurocholate (6 mg) and lyophilized Carbopol (3.5 mg) administered orally gave a sustained but comparatively small calcium lowering effect. In the in vitro enzymatic hydrolysis experiment, EC was more rapidly hydrolyzed in the intestinal fluid than in the mucosal extract. The combination of 20 mM taurocholate with 0.02% Carbopol showed the greatest inhibitory effect in both fluid and extract. These data indicated that EC was effectively absorbed through the intestinal wall, but the peptide was dominantly degraded by proteolytic enzymes in the GI tract. These results will offer a potential approach to the oral delivery of EC. PMID- 11411555 TI - Stealth polycyanoacrylate nanoparticles as tumor necrosis factor-alpha carriers: pharmacokinetics and anti-tumor effects. AB - The objective of this study was to investigate the pharmacokinetics and in vivo anti-tumor effect of recombinant human tumor necrosis factor-alpha (rHuTNF-alpha) encapsulated in poly(methoxypolyethyleneglycol cyanoacrylate-co-n-hexadecyl cyanoacrylate) (PEG-PHDCA) nanoparticles. Our experimental results showed that PEG-PHDCA nanoparticles could extend the half-life of rHuTNF-alpha to 7.42 h and obviously change the protein biodistribution in tissues, and in particular, increase accumulation of rHuTNF-alpha in tumor. Compared with PHDCA nanoparticles and free rHuTNF-alpha, PEG-PHDCA nanoparticles loaded with rHuTNF-alpha showed higher antitumor potency at the same dose, which might be related to its higher accumulation in tumor tissues and longer plasma circulation time. Therefore, PEG PHDCA nanoparticles could be an effective carrier for rHuTNF-alpha. PMID- 11411556 TI - PEGylated recombinant human tumor necrosis factor alpha: pharmacokinetics and anti-tumor effects. AB - The aim of the present work was to investigate and assess the merit of PEGylated recombinant human tumor necrosis factor-alpha (rHuTNF-alpha) following our previous work. The rHuTNF-alpha was modified using activated polyethylene glycol (PEG), N-succinimidyl succinnate monomethoxy polyethylene glycol (SS-PEG). The pharmacokinetics and anti-tumor effect were investigated. The experimental results showed that PEGylated rHuTNF-alpha could obviously alter in vivo behavioral characteristics of rHuTNF-alpha. Among the synthesized PEG-rHuTNF alphas with different PEG molecules, PEG20000-rHuTNF-alpha demonstrated the longest circulating half-life (24.8 h) which was about 50 times longer than that of rHuTNF-alpha (28.8 min). In addition, there was much more PEG20000-rHuTNF alpha distributed into tumor tissues than other PEG-rHuTNF-alphas or rHuTNF-alpha with time, and PEG20000-rHuTNF-alpha also showed the highest anti-tumor potency. These results indicated that PEG20000-rHuTNF-alpha was a useful long circulating molecule with selective localization in tumor tissues and enhanced anti-tumor activity of rHuTNF-alpha. PMID- 11411557 TI - Effect of lipophilicity on in vivo iontophoretic delivery. II. Beta-blockers. AB - The objective of this study was to investigate the relationship between drug lipophilicity and the transdermal absorption processes in the iontophoretic delivery in vivo. Anodal iontophoresis of beta-blockers as model drugs having different lipophilicity (atenolol, pindolol, metoprolol, acebutolol, oxprenolol and propranolol) was performed with rats (electrical current, 0.625 mA/cm2; application period, 90 min), and the drug concentrations in skin, cutaneous vein and systemic vein were determined. Increasing the lipophilicity of beta-blockers caused a greater absorption into the skin. Exceptionally, it was found that pindolol had high skin absorption, irrespective of its hydrophilic nature. Further, the drug transfer rate from skin to cutaneous vein (R(SC)) was evaluated from the arterio-venous plasma concentration difference of drug in the skin. Normalized R(SC) by skin concentration showed a negative correlation with the logarithm of n-octanol/buffer partition coefficient (Log P, pH 7.4), suggesting the partitioning between stratum corneum and viable epidermis was a primary process to determine the transfer properties of beta-blockers to local blood circulation. Pindolol exhibited both high skin absorption and high transfer from skin to cutaneous vein. These characteristics of pindolol could be explained by the chemical structure, molecular size and hydrophilicity. These findings for pindolol should be valuable for the optimal design of drug candidates for iontophoretic transdermal delivery. PMID- 11411558 TI - Metabolism of ipecac alkaloids cephaeline and emetine by human hepatic microsomal cytochrome P450s, and their inhibitory effects on P450 enzyme activities. AB - In this study, we identified the metabolites and the CYP forms that are specifically involved in emetine O-demethylation in human liver microsomes, and cleared the inhibitory potential of cephaeline and emetine on the activity of the major drug-metabolizing CYP enzymes. Incubation of emetine with human liver microsomes yielded three metabolites identified by using HPLC by comparison of the retention time with the authentic sample of cephaeline, 9-O-demethylemetine and 10-O-demethylemetine. CYP3A4 and CYP2D6 were able to metabolize emetine to cephaeline and 9-O-demethylemetine, and CYP3A4 also participated in metabolizing emetine to 10-O-demethylemetine. Cephaeline and emetine inhibited probe substrates metabolism. IC50 for cephaeline against CYP2D6 and CYP3A4 were 121 and 1000 microM, respectively. For the emetine, CYP2D6 and CYP3A4 were 80 and 480 microM, respectively. Inhibition constants (Ki) for both compounds on the CYP2D6 and CYP3A4 activities were determined by graphic analysis of Dixon plots at various concentrations. The obtained Ki values of cephaeline for CYP2D6 and CYP3A4 were 54 and 355 microM, respectively, and the values of emetine were 43 and 232 microM, respectively. We concluded that these in vitro inhibitions of cephaeline and emetine would hardly increase plasma concentrations of co administered drugs in clinical therapy. PMID- 11411559 TI - Effect of serum triglyceride concentration on the fluctuation of whole blood concentration of cyclosporin A in patients. AB - The methodology to distinguish the patients showing considerable fluctuation of the whole blood concentration of cyclosporin A (CYA) was investigated from a viewpoint of laboratory test values. First, we retrospectively examined the CYA trough blood concentrations monitored continuously. The patients were classified into three groups by the fluctuation of CYA trough blood concentrations during the examination period (Cmax/Cmin): Group 1 (Cmax/Cmin=100-200%; n=21), Group 2 (Cmax/Cmin=200-300%; n=25), and Group 3 (Cmax/Cmin=more than 300%; n=32). In the laboratory tests examined, the serum triglyceride concentrations were considerably different among the groups, and it was the highest in Group 3. Next, to elucidate the effect of serum triglyceride concentration on the CYA blood concentration, in vivo pharmacokinetic studies after single intravenous or repetitive oral administration of CYA were conducted in the model rats with pseudo-hypertriglyceridemia, hypocythemia, and acute renal failure. Only in pseudo-hypertriglyceridemia rats, the CYA blood concentration after a single intravenous injection was significantly higher than that in normal rats because of the restriction of CYA distribution to the extravascular tissues. On the other hand, the increase in the serum triglyceride concentration did not affect the fluctuation of CYA trough blood concentration after repetitive oral administration. Taken together, the fluctuation of CYA trough blood concentrations observed in the clinical situation could be due to the fluctuation of serum triglyceride concentration, and the patients with such fluctuation of serum triglyceride concentrations might also be distinguishable by the higher concentration of serum triglyceride in laboratory tests. PMID- 11411560 TI - The inhibitory effect of an extract of Sanguisorba officinalis L. on ultraviolet B-induced pigmentation via the suppression of endothelin-converting enzyme 1alpha. AB - Endothelin-1 (ET-1) has been reported to be expressed in human epidermis at both the gene and protein levels. ET-1 plays a pivotal role in ultraviolet B (UVB) induced pigmentation due to its accentuated secretion after UVB irradiation and its function as a mitogen and as a melanogen for human melanocytes. We have recently found that endothelin-converting enzyme (ECE)-1alpha plays a constitutive role in the secretion of ET-1 by human keratinocytes and that an extract of Sanguisorba officinalis L. inhibits ECE activity in human endothelial cells, which predominantly express ECE-1alpha. In this report, to clarify the potential use of this botanical extract as a whitening agent, we examined whether this extract inhibits UVB-induced pigmentation in vivo. When this extract was applied to human keratinocytes after UVB irradiation, secretion of ET-1 by those cells was reduced, and this was accompanied by a concomitant increase in the secretion of inactive precursor Big endothelin-1. When hairless mice were exposed to UVB light and were treated with the extract, it suppressed the induction of ET 1 in the UVB-irradiated epidermis. In the course of UVB-induced pigmentation of brownish guinea pig skin, this extract significantly diminished pigmentation in UVB-exposed areas. These findings indicate that ECE-1alpha in keratinocytes plays a pivotal role in the induction of pigmentation following UVB irradiation and that an extract of S. officinalis, which inhibits ET-1 production in human keratinocytes, is a good ingredient for a whitening agent. PMID- 11411561 TI - Determination of 14 chemical constituents in the traditional Chinese medicinal preparation Huangqin-Tang by high performance liquid chromatography. AB - The high performance liquid chromatographic (HPLC) method for the identification and determination of baicalin (BG), wogonoside (WG), oroxylin-A-glucoside (OG), baicalein (B), wogonin (W), orxylin-A (O), paeoniflorin (PF), glycyrrhizic acid (GL), glycyrrhetinic acid (GA), liquiritin (LG), isoliquirition (ILG), liquiritigenin (L), isoliquiritigenin (IL) and ononin (ON) in Huangqin-Tang [Chinese characters: see text] was established. The samples were separated with a Wakosil C18 column (4.6 x 150 mm) by linear gradient elution using A (MeOH-HAC 100:1, v/v)-B (Water-HAC 100:1, v/v) (0 min, 30:70; 15 min, 40:60; 30 min, 60:40; 45 min, 80:20; 60 min, 100:0) as the mobile phase at a flow-rate of 1.0 ml/min. The detection was by diode-array UV/Vis detector (DAD), and the wavelength was set at the range of 200-400 nm. Satisfactory results were obtained within 60 min for the simultaneous determination of the 14 constituents. The repeatability (RSD) of the method was generally less than 2% (n=5, interday and intraday). The recovery of BG was 96.9+/-1.71, WG was 98.9+/-2.99, PF was 99.7+/-0.52, LG was 95.3+/-2.67, GL was 96.7+/-3.44, and GA was 94.8+/-4.16, respectively. PMID- 11411562 TI - Enhancing effect of alpha-monoisostearyl glyceryl ether on the percutaneous penetration of indomethacin through excised rat skin. AB - The enhancing effect of alpha-monoisostearyl glyseryl ether (GE-IS) on the percutaneous penetration of indomethacin (IM) from test solutions in propylene glycol (PG) was investigated using the excised abdominal skin of rats in vitro. The percutaneous penetration of IM into diffusion cells was significantly increased in the presence of 0.2% or 1% (w/w) GE-IS compared with enhancer-free PG solution. Permeation parameters of IM, such as lag time and permeability coefficient, revealed that GE-IS significantly augmented the percutaneous penetration of IM from PG. These results strongly suggested that GE-IS functions as a penetration enhancer of IM through rat skin. To elucidate the mode of action of GE-IS as a penetration enhancer, the solubility of IM in the test solution and the percutaneous penetration of IM through damaged skin from which the stratum corneum had been stripped were investigated. The results suggested that GE-IS acts directly on the stratum corneum and alters the permeability of the skin. PMID- 11411563 TI - Inhibitory effects of anti-inflammatory drugs on interleukin-6 bioactivity. AB - Interleukin-6 (IL-6) is known as a proinflammatory cytokine involved in immune response, inflammation, and hematopoiesis. Inhibitory effects of anti inflammatory drugs on IL-6 bioactivity using IL-6-dependent hybridoma have been evaluated. Three out of 16 nonsteroidal anti-inflammatory drugs (NSAIDs) showed IC50 values of less than 100 microM, which were in the order of oxyphenylbutazone hydrate (IC50=7.5 microM)>meclofenamic acid sodium salt (31.9 microM)>sulindac (74.9 microM). Steroidal anti-inflammatory drugs (SAIDs) exhibited significant inhibitory effects at 100 microM on the IL-6 bioactivity, and their inhibitory potencies were in the order of budesonide (IC50=2.2 microM)>hydrocortisone 21 hemisuccinate (6.7 microM), prednisolone (7.5 microM), betamethasone (10.9 microM)>dexamethasone (18.9 microM) and triamcinolone acetonide (24.1 microM). The results would provide an additional mechanism by which anti-inflammatory drugs display their anti-inflammatory and immunosuppressive effects at higher concentrations. PMID- 11411564 TI - DNA-binding activity and cytotoxicity of the extended diphenylfuran bisamidines in breast cancer MCF-7 cells. AB - The DNA binding properties of three novel extended diphenylfuran bisamidines (1 3) possessing different dicationic terminal side chains were studied. The ultrafiltration assay showed that bisamidines 1-3 have significant affinity for DNA. The DNA-binding data for bisamidines 1-3 using homopolymers poly(dA-dT)- poly(dA-dT) and poly(dG-dC)- poly(dG-dC), indicated that these compounds show moderate specificity for AT base pairs. We studied the cytotoxicity effects of bisamidines 1-3, Hoechst 33258 and DAPI (4',6-diamidino-2-phenylindole) in cultured breast cancer MCF-7 cells. The bisamidines 1-3 showed comparable antitumour activity to Hoechst 33258, but were substantially more cytotoxic compared to DAPI. These data show that in broad terms the cytotoxic potency of bisamidines 1-3 in cultured breast cancer MCF-7 cells decreases with the size of the alkyl group substituent (cyclopropyl>isopropyl>cyclopentyl), in accord with their increases in DNA affinity, as shown by the binding constant values. PMID- 11411565 TI - Hydrolyzable tannins, the active constituents of three Greek Cytinus taxa against several tumor cell lines. AB - Hydrolyzable tannins were found to be the active cytotoxic constituents of three Greek Cytinus taxa: Cytinus ruber, Cytinus hypocistis subsp. hypocistis and Cytinus hypocistis subsp. orientalis. The cytotoxic activity was evaluated against a broad spectrum of cancer cell lines. The structure of the active compounds was investigated with NMR and electrospray-MS/MS techniques. PMID- 11411566 TI - The effect of ginkgo biloba extract on free radical production in hypoxic rats. AB - In the present study, we assayed the antioxidant properties of Ginkgo biloba (Gb) extract on rats submitted to 21 d of chronic hypoxia. Doses of 25 and 50 mg/kg were examined. Oxygenated free radical production measured by the chemiluminescence technique was significantly decreased in treated rats compared to control rats placed in similar experimental conditions, and this effect was more significant at the 50 mg/kg dose. On the other hand, no antioxidant enzyme activities of the drug were observed towards red blood cells. These results suggest that ginkgo biloba extract has a free radical scavenging action. These antioxidant properties could explain the beneficial hematological properties of Gb extract. PMID- 11411567 TI - Effect of administration with the extract of Gymnema sylvestre R. Br leaves on lipid metabolism in rats. AB - Extract of Gymnema sylvestre R. Br leaves (GE) was orally administered once a day to rats fed a high fat diet or normal fat diet for 3 weeks to investigate its influence on lipid metabolism. As a result, GE did not influence body weight gain or feed intake in both diet groups during the experimental period. The apparent fat digestibility was significantly decreased by GE in both diet groups for the last 2 weeks of the experimental period, though not the apparent protein digestibility. In addition, the excretion of neutral sterols and acid steroids into feces was increased by GE in both diet groups. Furthermore, GE decreased the total cholesterol and triglyceride levels in serum. On the other hand, blood lecithin-cholesterol acyltransferase (LCAT) activity was increased by GE. Moreover, it was suggested that GE influenced cecal fermentation and that propionic acid and acetic acid contents in cecum were significantly increased by GE. Consequently, it was suggested that GE improved serum cholesterol and triglyceride levels through influence over a wide range of lipid metabolism in rats. PMID- 11411568 TI - Antifungal activity of modified hederagenin glycosides from the leaves of Kalopanax pictum var. chinense. AB - Monodesmosides which were obtained from the partial degradation of hederagenin bisdesmosides exhibited significant antifungal effect against Microsporum canis, Coccidioides immitis, Trichophyton mentagrophytes, Cryptococcus neoformans, and Candida albicans at the minimal inhibitory concentrations of 6.25-25 microg/ml. The hederagenin glycosides were isolated from the leaves of Kalopanax pictum var. chinense. PMID- 11411569 TI - Screening of natural compounds for inhibitory activity on colon cancer cell migration. AB - We examined the effects of 75 kinds of natural compounds, such as alkaloids, phenylpropanoids, flavonoids, steroids and terpenoids on the in vitro migration and proliferation of colon 26-L5 cells, in comparison with anti-cancer drugs used for chemotherapy. Twenty-three of the 75 compounds inhibited markedly tumor cell migration. Among the 23 compounds, evodiamine showed the most potent and selective inhibitory activity on tumor cell migration with an IC50 value of 1.25 microg/ml, which was about 20 times lower than that for tumor cell proliferation. The migratory inhibition reached about 70% at 10 microg/ml of evodiamine. On the other hand, most of anti-cancer drugs tested, except for paclitaxel, had little effect on tumor cell migration at the concentrations strongly inhibiting tumor cell proliferation. Paclitaxel suppressed tumor cell migration in a concentration dependent manner and achieved about 70% inhibition at 10 microg/ml with a marginal effect on cell proliferation. These results suggest that evodiamine and paclitaxel may be regarded as leading compounds for anti-metastatic agents acting through the inhibition of tumor cell migration without cytotoxicity. PMID- 11411570 TI - Adsorptive capacity of activated charcoal for rifampicin with and without sodium chloride and sodium citrate. AB - The effects of two saline cathartics (sodium chloride and sodium citrate) on the adsorptive capacity of activated charcoal (AC) for rifampicin were studied. Solutions of rifampicin alone and rifampicin with 7.5 mg/ml cathartic solution were vortex-mixed for 30 s with different quantities of AC. These were incubated for 30 min at 37 degrees C and analyzed for free rifampicin spectrophotometrically at 320 nm. The addition of sodium citrate significantly increased (p<0.05) the adsorptive capacity of AC for rifampicin with a resulting decrease in B-50 values at both the therapeutic and simulated toxic doses. Sodium chloride addition reduced the binding of rifampicin to AC at the toxic doses. The adsorption of rifampicin to activated charcoal, both alone and with the two saline cathartics, obeyed quantity-dependent kinetics. AC may be co-administered with sodium citrate in the management of rifampicin overdose. PMID- 11411571 TI - 2,2-Diphenyl-1-picrylhydrazyl hydrate, a stable free radical, is an alpha glucosidase inhibitor. AB - Glycosidases play a pivotal role in processing of various glycoproteins and glycolipids. It is well known that glycosidases are also involved in a variety of degenerative metabolic disorders such as cancer and AIDS. In order to develop potent alpha-glucosidase inhibitors, we first screened 2,2-diphenyl-1 picrylhydrazyl hydrate as a candidate. 2,2-Diphenyl-1-picrylhydrazyl hydrate was found to inhibit alpha- and beta-glucosidases as well as alpha- and beta mannosidases. It was also shown to be a non-competitive inhibitor of yeast alpha glucosidase with a Ki value of 1.1 x 10(-6) M. Taken together, we anticipate that 2,2-diphenyl-1-picrylhydrazyl hydrate may be a potent inhibitor for some incurable metabolic disorders including AIDS. PMID- 11411572 TI - Possible existence of a novel receptor for uridine analogues in the central nervous system using two isomers, N3-(S)-(+)- and N3-(R)-(-)-alpha-hydroxy-beta phenethyluridines. AB - Uridine analogue binding sites, the so-called uridine receptor, were observed in the experiments on specific [3H]N3-phenacyluridine binding to bovine synaptic membranes using two isomers, N3-(S)-(+)- and N3-(R)-(-)-alpha-hydroxy-beta phenethyluridine, as ligands. The potent hypnotic, N3-(S)-(+)-alpha-hydroxy-beta phenethyluridine, but not the (R)-isomer, strongly inhibited [3H]N3 phenacyluridine binding. The racemate had inhibitory activity intermediate between that of the two alpha-hydroxy-beta-phenethyluridines ((R)- or (S) isomers). The inhibitory constants of these compounds were determined. The Ki values of N3-phenacyluridine, alpha-hydroxy-beta-phenethyluridine (racemate), N3 (R)-(-)-, and N3-(S)-(+)-alpha-hydroxy-beta-phenethyluridine were 0.65, 397.4, 1908, and 10.2 nM, respectively. The present results indicate the existence of uridine receptors in the central nervous system in relation to their hypnotic activities reported previously. PMID- 11411573 TI - Histamine production in mouse epidermal keratinocytes is regulated during cellular differentiation. PMID- 11411574 TI - Histamine as a potential autocrine regulator of melanoma. PMID- 11411575 TI - Histamine synthesis by mouse mammary gland epithelial cells in primary culture. The effects of mammary gland differentiation stage (pregnancy and lactation). PMID- 11411576 TI - Comparison of ligand affinity values at the recombinant human H3 receptor and at H3 receptor(s) of human cingulate cortex. PMID- 11411577 TI - Effects of (R)alpha-methylhistamine on experimental colitis. PMID- 11411578 TI - Role of K+ in the histamine H3 receptor- and adenosine A1 receptor-mediated effects in the guinea-pig isolated duodenum. PMID- 11411579 TI - Intracellular histamine content increases during in vitro dendritic cell differentiation. PMID- 11411580 TI - Gastroprotection by (R)-alpha-methylhistamine: involvement of nitric oxide? PMID- 11411581 TI - Gastroprotective activity of the novel histamine H3-receptor agonist FUB 407. PMID- 11411582 TI - Hydrogen peroxide and the response of trachea of sensitized guinea-pigs: opposite effects on contraction and histamine release. PMID- 11411583 TI - Role of the endothelium in the vasorelaxing effect of histamine on rat isolated gastric small arteries. PMID- 11411584 TI - Relaxin generates nitric oxide and provides protection against cardiac anaphylaxis. PMID- 11411585 TI - The heart protective/damaging effects of short/long lasting mesenteric ischaemia may be mimicked by exogenous histamine (II). PMID- 11411586 TI - Structure and function of copper-containing amine oxidases. PMID- 11411587 TI - Selective inhibition of FAD and copper-dependent amine oxidases by N-acetyl pyrazole derivatives. PMID- 11411588 TI - Analysis of the expression of the murine diamine oxidase gene. PMID- 11411589 TI - Role of monoamine oxidases on rat liver mitochondrial function. PMID- 11411590 TI - Enrichment of a human leukemia cell line (K562) with a plant histaminase. PMID- 11411591 TI - Inhibition of monoamine oxidase by metronidazole. PMID- 11411592 TI - A study on the reactions of plant copper amine oxidase with short-chain aliphatic diamines. PMID- 11411593 TI - Histamine weakly stimulates lipolysis and is poorly oxidized by amine oxidases in human subcutaneous fat cells. PMID- 11411594 TI - Substrates of semicarbazide-sensitive amine oxidase mimic diverse insulin effects in adipocytes. PMID- 11411595 TI - Some properties of semicarbazide-sensitive amine oxidase (SSAO) from human cerebrovascular tissues. PMID- 11411596 TI - Detection of histamine and other biogenic amines using biosensors based on amine oxidase. PMID- 11411597 TI - Inhibition of histamine release of human basophils and mast cells in vitro by ultraviolet A (UVA) irradiation. PMID- 11411598 TI - Flow-cytometric analysis of basophil activation: inhibition by histamine at conventional and homeopathic concentrations. PMID- 11411599 TI - Effects of antioxidant vitamins on anti-IgE-induced mediator release from human basophils. PMID- 11411600 TI - Effect of chloride channel blockers on anti-IgE-stimulated histamine and IL-4/IL 13 release from human basophils. PMID- 11411601 TI - PI 3-kinase and MEK kinase inhibitors differentially affect IgE-dependent and IgE independent basophil mediator release. PMID- 11411602 TI - Highly reduced peritoneal mast cell number and decreased c-kit expression in histidine decarboxylase knock out mice. PMID- 11411603 TI - Protease activated receptors and mast cell activation. PMID- 11411604 TI - The dose-dependent release of histamine from placental mast cells after administration of atrial natriuretic peptide. PMID- 11411605 TI - Modulation of intracellular cyclic AMP in immunologically activated rat peritoneal mast cells by prostaglandin D2. PMID- 11411606 TI - Stimulatory effect of high dilutions of histamine on activation of human basophils induced by anti-IgE. PMID- 11411607 TI - Plasma histamine levels during plasmapheresis: difficult interpretation of adverse reactions to plasma substitutes. PMID- 11411608 TI - Analysis of diamine oxidase gene polymorphisms in patients with inflammatory bowel disease. PMID- 11411609 TI - The relationship between body size and the urinary excretion of the main histamine metabolite tele-methylimidazoleacetic acid in man. PMID- 11411610 TI - Histamine neurons down-regulate ob gene expression in rat white adipose tissue. PMID- 11411611 TI - Modulation of acetylcholine release by histamine in the nucleus accumbens. PMID- 11411612 TI - Central histaminergic H3-receptors and locomotor activity. PMID- 11411613 TI - Cognitive enhancing properties of thioperamide infused into the nucleus basalis magnocellularis of the rat. PMID- 11411614 TI - Histamine H1-antagonists potentiate seizures in the EL (epilepsy-like) mouse model of temporal lobe epilepsy. PMID- 11411615 TI - Effect of amthamine, a histamine H2-agonist, in two mouse chemical-induced seizure models. PMID- 11411616 TI - The influence of histamine H1-, H2- and H3-receptor antagonists on histamine stimulated NGF release from cultured astrocytes. PMID- 11411617 TI - Hepatocyte transplantation reverses the effect of portocaval shunt on rat brain histamine. PMID- 11411618 TI - Characterization by flow cytometry of hemopoietic progenitors in bone marrow of histidine decarboxylase knock out and wild type mice. PMID- 11411619 TI - Detection of histamine and histidine decarboxylase in human platelets by flow cytometry. PMID- 11411620 TI - Inhibitory effect of ascorbic acid on human retinal pigment epithelial cell proliferation compared to cytostatic drugs--influence of histamine. PMID- 11411621 TI - Determination of histamine degradation capacity in extremely small human colon samples. PMID- 11411622 TI - Intracellular localization of histidine decarboxylase. PMID- 11411623 TI - Influence of compression therapy on symptoms following soft tissue injury from maximal eccentric exercise. AB - STUDY DESIGN: A between groups design was used to compare recovery following eccentric muscle damage under 2 experimental conditions. OBJECTIVE: To determine if a compression sleeve donned immediately after maximal eccentric exercise would enhance recovery of physical function and decrease symptoms of soreness. BACKGROUND: Prior investigations using ice, intermittent compression, or exercise have not shown efficacy in relieving symptoms of delayed onset muscle soreness (DOMS). To date, no study has shown the effect of continuous compression on DOMS, yet this would offer a low cost intervention for patients suffering with the symptoms of DOMS. METHODS AND MEASURES: Twenty nonimpaired non-strength-trained women participated in the study. Subjects were matched for age, anthropometric data, and one repetition maximum concentric arm curl strength and then randomly placed into a control group (n = 10) or an experimental compression sleeve group (n = 10). Subjects were instructed to avoid pain-relieving modalities (eg, analgesic medications, ice) throughout the study. The experimental group wore a compressive sleeve garment for 5 days following eccentric exercise. Subjects performed 2 sets of 50 passive arm curls with the dominant arm on an isokinetic dynamometer with a maximal eccentric muscle action superimposed every fourth passive repetition. One repetition maximum elbow flexion, upper arm circumference, relaxed elbow angle, blood serum cortisol, creatine kinase, lactate dehydrogenase, and perception of soreness questionnaires were collected prior to the exercise bout and daily thereafter for 5 days. RESULTS: Creatine kinase was significantly elevated from the baseline value in both groups, although the experimental compression test group showed decreased magnitude of creatine kinase elevation following the eccentric exercise. Compression sleeve use prevented loss of elbow motion, decreased perceived soreness, reduced swelling, and promoted recovery of force production. CONCLUSIONS: Results from this study underline the importance of compression in soft tissue injury management. PMID- 11411624 TI - Structure and function of the lumbar intervertebral disk in health, aging, and pathologic conditions. AB - This report is a comprehensive review of the basic and clinical science relating to the morphology and function of the intervertebral disc of the lumbar spine. The purpose is to review the anatomy, physiology, and biomechanics of the intervertebral disc of the lumbar spine in health, with aging, and in pathologic conditions. The complex morphology and ultrastructure of the intervertebral disc of the lumbar spine in the human provide the critical elements that permit normal mobility and transmission of force through the vertebral column. Alterations in this structure are manifest in a variety of clinical conditions routinely encountered in orthopaedic physical therapy practice. These structural and biomechanical changes are related to degenerative changes that occur in association with aging and trauma. Knowledge of the gross morphology and ultrastructure of the intervertebral disc and pathobiologic processes underlying associated conditions is essential to orthopaedic practice. PMID- 11411625 TI - Understanding statistical power. AB - This article provides an introduction to power analysis so that readers have a basis for understanding the importance of statistical power when planning research and interpreting the results. A simple hypothetical study is used as the context for discussion. The concepts of false findings and missed findings are introduced as a way of thinking about type I and type II errors. The primary factors that affect power are described and examples are provided. Finally, examples are presented to demonstrate 2 uses of power analysis, 1 for prospectively estimating the sample size needed to insure finding effects of a known magnitude in a study and 1 for retrospectively estimating power to gauge the likelihood that an effect was missed. PMID- 11411626 TI - Effect of soft lumbar support belt on abdominal oblique muscle activity in nonimpaired adults during squat lifting. AB - STUDY DESIGN: A counterbalanced repeated measures design. OBJECTIVE: To determine the effect of a soft lumbar support on abdominal oblique muscle electromyography (EMG) during lifting. BACKGROUND: Use of a soft lumbar back support is a common preventive measure to reduce the incidence of back injuries. Because the abdominal oblique muscles stabilize the spine during lifting, wearing this support may alter the activity level of these muscles. METHODS AND MEASURES: Twenty nonimpaired subjects (14 women and 6 men, mean age 28.9 +/- 8.1 years) were tested using surface electrodes to record the EMG of the right abdominal oblique muscles during lifting with and without the support. RESULTS: A paired t test revealed a significant decrease in activity of the abdominal oblique muscles during lifting with the support (29.7 +/- 3.13 mV) compared to without it (33.3 +/- 3.05 mV). Of the 20 subjects, the 14 women showed decreased EMG amplitude during the lift with the lumbar support compared to without the support, while 5 of the men showed increased EMG amplitude with the support and 1 man showed no change. CONCLUSION: This finding suggests that the soft lumbar support may play some role in the stabilization of the lumbar region during lifting. PMID- 11411627 TI - Quantitative examinations for multi joint arm trajectory planning--using a robust calculation algorithm of the minimum commanded torque change trajectory. AB - In previous research, criteria based on optimal theories were examined to explain trajectory features in time and space in multi joint arm movement. Four criteria have been proposed. They were the minimum hand jerk criterion (by which a trajectory is planned in an extrinsic-kinematic space), the minimum angle jerk criterion (which is planned in an intrinsic-kinematic space), the minimum torque change criterion (where control objects are joint links; it is planned in an intrinsic-dynamic-mechanical space), and the minimum commanded torque change criterion (which is planned in an intrinsic space considering the arm and muscle dynamics). Which of these is proper as a criterion for trajectory planning in the central nervous system has been investigated by comparing predicted trajectories based on these criteria with previously measured trajectories. Optimal trajectories based on the two former criteria can be calculated analytically. In contrast, optimal trajectories based on the minimum commanded torque change criterion are difficult to be calculated, even with numerical methods. In some cases, they can be computed by a Newton-like method or a steepest descent method combined with a penalty method. However, for a realistic physical parameter range, the former becomes unstable quite often and the latter is unreliable about the optimality of the obtained solution. In this paper, we propose a new method to stably calculate optimal trajectories based on the minimum commanded torque change criterion. The method can obtain trajectories satisfying Euler-Poisson equations with a sufficiently high accuracy. In the method, a joint angle trajectory, which satisfies the boundary conditions strictly, is expressed by using orthogonal polynomials. The coefficients of the orthogonal polynomials are estimated by using a linear iterative calculation so as to satisfy the Euler Poisson equations with a sufficiently high accuracy. In numerical experiments, we show that the optimal solution can be computed in a wide work space and can also be obtained in a short time compared with the previous methods. Finally, we perform supplementary examinations of the experiments by Nakano, Imamizu, Osu, Uno, Gomi, Yoshioka et al. (1999). Estimation of dynamic joint torques and trajectory formation from surface electromyography signals using a neural network model. Biological Cybernetics, 73, 291-300. Their experiments showed that the measured trajectory is the closest to the minimum commanded torque change trajectory by statistical examination of many point-to-point trajectories over a wide range in a horizontal and sagittal work space. We recalculated the minimum commanded torque change trajectory using the proposed method, and performed the same examinations as previous investigations. As a result, it could be reconfirmed that the measured trajectory is closest to the minimum commanded torque change trajectory previously reported. PMID- 11411628 TI - Solving the binding problem of the brain with bi-directional functional connectivity. AB - We propose a neural network model which gives one solution to the binding problem on the basis of 'functional connectivity' and bidirectional connections. Here, 'functional connectivity' is dynamic neuronal connectivity peculiar to temporal spike coding neural networks with coincidence detector neurons. The model consists of a single primary map and two higher modules which extract two different features shown on the primary map. There exist three layers in each higher module and the layers are connected bi-directionally. An object in the outer world is represented by a 'global dynamical cell assembly' which is organized across the primary map and the two higher modules. Detailed, but spatially localized, information is coded in the primary map, whereas coarse, but spatially extracted information or globally integrated information is coded in the higher modules. Computer simulations of the proposed model show that multiple cell assemblies sharing the same neurons partially can co-exist. Furthermore, we introduce a three-dimensional J-PSTH (Joint-Peri Stimulus Time Histogram) which is capable of tracking such cell assemblies, altering its constituent neurons as in our proposed model. PMID- 11411629 TI - Learning from noisy information in FasArt and FasBack neuro-fuzzy systems. AB - Neuro-fuzzy systems have been in the focus of recent research as a solution to jointly exploit the main features of fuzzy logic systems and neural networks. Within the application literature, neuro-fuzzy systems can be found as methods for function identification. This approach is supported by theorems that guarantee the possibility of representing arbitrary functions by fuzzy systems. However, due to the fact that real data are often noisy, generation of accurate identifiers is presented as an important problem. Within the Adaptive Resonance Theory (ART), PROBART architecture has been proposed as a solution to this problem. After a detailed comparison of these architectures based on their design principles, the FasArt and FasBack models are proposed. They are neuro-fuzzy identifiers that offer a dual interpretation, as fuzzy logic systems or neural networks. FasArt and FasBack can be trained on noisy data without need of change in their structure or data preprocessing. In the simulation work, a comparative study is carried out on the performances of Fuzzy ARTMAP, PROBART, FasArt and FasBack, focusing on prediction error and network complexity. Results show that FasArt and FasBack clearly enhance the performance of other models in this important problem. PMID- 11411630 TI - Comparing Bayesian neural network algorithms for classifying segmented outdoor images. AB - In this paper we investigate the Bayesian training of neural networks for region labelling of segmented outdoor scenes; the data are drawn from the Sowerby Image Database of British Aerospace. Neural networks are trained with two Bayesian methods, (i) the evidence framework of MacKay (1992a,b) and (ii) a Markov Chain Monte Carlo method due to Neal (1996). The performance of the two methods is compared to evaluating the empirical learning curves of neural networks trained with the two methods. We also investigate the use of the Automatic Relevance Determination method for input feature selection. PMID- 11411631 TI - Three learning phases for radial-basis-function networks. AB - In this paper, learning algorithms for radial basis function (RBF) networks are discussed. Whereas multilayer perceptrons (MLP) are typically trained with backpropagation algorithms, starting the training procedure with a random initialization of the MLP's parameters, an RBF network may be trained in many different ways. We categorize these RBF training methods into one-, two-, and three-phase learning schemes. Two-phase RBF learning is a very common learning scheme. The two layers of an RBF network are learnt separately; first the RBF layer is trained, including the adaptation of centers and scaling parameters, and then the weights of the output layer are adapted. RBF centers may be trained by clustering, vector quantization and classification tree algorithms, and the output layer by supervised learning (through gradient descent or pseudo inverse solution). Results from numerical experiments of RBF classifiers trained by two phase learning are presented in three completely different pattern recognition applications: (a) the classification of 3D visual objects; (b) the recognition hand-written digits (2D objects); and (c) the categorization of high-resolution electrocardiograms given as a time series (ID objects) and as a set of features extracted from these time series. In these applications, it can be observed that the performance of RBF classifiers trained with two-phase learning can be improved through a third backpropagation-like training phase of the RBF network, adapting the whole set of parameters (RBF centers, scaling parameters, and output layer weights) simultaneously. This, we call three-phase learning in RBF networks. A practical advantage of two- and three-phase learning in RBF networks is the possibility to use unlabeled training data for the first training phase. Support vector (SV) learning in RBF networks is a different learning approach. SV learning can be considered, in this context of learning, as a special type of one phase learning, where only the output layer weights of the RBF network are calculated, and the RBF centers are restricted to be a subset of the training data. Numerical experiments with several classifier schemes including k-nearest neighbor, learning vector quantization and RBF classifiers trained through two phase, three-phase and support vector learning are given. The performance of the RBF classifiers trained through SV learning and three-phase learning are superior to the results of two-phase learning, but SV learning often leads to complex network structures, since the number of support vectors is not a small fraction of the total number of data points. PMID- 11411632 TI - Noise suppression in training examples for improving generalization capability. AB - For the supervised learning problem, error correcting memorization learning was proposed in order to suppress noise in teacher signals. In this paper, generalization capability of the learning method is discussed. Generalization capability is evaluated based on the projection learning criterion. We give a necessary and sufficient condition for error correcting memorization learning to provide the same level of generalization as projection learning, and suggest how to choose a training set so as to satisfy the obtained condition. Moreover, it is revealed that noise suppression based on the error correcting memorization learning criterion always has a good effect on improving generalization to the level of projection learning. PMID- 11411633 TI - Networks with trainable amplitude of activation functions. AB - Network training algorithms have heavily concentrated on the learning of connection weights. Little effort has been made to learn the amplitude of activation functions, which defines the range of values that the function can take. This paper introduces novel algorithms to learn the amplitudes of nonlinear activations in layered networks, without any assumption on their analytical form. Three instances of the algorithms are developed: (i) a common amplitude is shared among all nonlinear units; (ii) each layer has its own amplitude; and (iii) neuron-specific amplitudes are allowed. The algorithms can also be seen as a particular double-step gradient-descent procedure, as gradient-driven adaptive learning rate schemes, or as weight-grouping techniques that are consistent with known scaling laws for regularization with weight decay. As a side effect, a self pruning mechanism of redundant neurons may emerge. Experimental results on function approximation, classification, and regression tasks, with synthetic and real-world data, validate the approach and show that the algorithms speed up convergence and modify the search path in the weight space, possibly reaching deeper minima that may also improve generalization. PMID- 11411634 TI - A model with an intrinsic property of learning higher order correlations. AB - A neural network model that can learn higher order correlations within the input data without suffering from the combinatorial explosion problem is introduced. The number of parameters scales as M x N, where M is the number such that no higher order network with less than M higher order terms can implement the same input data set and N is the dimensionality of the input vectors. In order to have better generalization, the model was designed to realize a supervised learning such that after learning, output for any input vector is the same as the output of a higher order network that implements the same input data set using M number of higher order terms. Unlike the case in product units, the local minima problem does not pose itself as a severe problem in the model. Simulation results for some problems are presented and the results are compared with the results of a multilayer feedforward network. It is observed that the model can generalize better than the multilayer feedforward network. PMID- 11411635 TI - S-TREE: self-organizing trees for data clustering and online vector quantization. AB - This paper introduces S-TREE (Self-Organizing Tree), a family of models that use unsupervised learning to construct hierarchical representations of data and online tree-structured vector quantizers. The S-TREE1 model, which features a new tree-building algorithm, can be implemented with various cost functions. An alternative implementation, S-TREE2, which uses a new double-path search procedure, is also developed. The performance of the S-TREE algorithms is illustrated with data clustering and vector quantization examples, including a Gauss-Markov source benchmark and an image compression application. S-TREE performance on these tasks is compared with the standard tree-structured vector quantizer (TSVQ) and the generalized Lloyd algorithm (GLA). The image reconstruction quality with S-TREE2 approaches that of GLA while taking less than 10% of computer time. S-TREE1 and S-TREE2 also compare favorably with the standard TSVQ in both the time needed to create the codebook and the quality of image reconstruction. PMID- 11411636 TI - The constraint based decomposition (CBD) training architecture. AB - The Constraint Based Decomposition (CBD) is a constructive neural network technique that builds a three or four layer network, has guaranteed convergence and can deal with binary, n-ary, class labeled and real-value problems. CBD is shown to be able to solve complicated problems in a simple, fast and reliable manner. The technique is further enhanced by two modifications (locking detection and redundancy elimination) which address the training speed and the efficiency of the internal representation built by the network. The redundancy elimination aims at building more compact architectures while the locking detection aims at improving the training speed. The computational cost of the redundancy elimination is negligible and this enhancement can be used for any problem. However, the computational cost of the locking detection is exponential in the number of dimensions and should only be used in low dimensional spaces. The experimental results show the performance of the algorithm presented in a series of classical benchmark problems including the 2-spiral problem and the Iris, Wine, Glass, Lenses, Ionosphere, Lung cancer, Pima Indians, Bupa, TicTacToe, Balance and Zoo data sets from the UCI machine learning repository. CBD's generalization accuracy is compared with that of C4.5, C4.5 with rules, incremental decision trees, oblique classifiers, linear machine decision trees, CN2, learning vector quantization (LVQ), backpropagation, nearest neighbor, Q* and radial basis functions (RBFs). CBD provides the second best average accuracy on the problems tested as well as the best reliability (the lowest standard deviation). PMID- 11411637 TI - Life-long learning cell structures--continuously learning without catastrophic interference. AB - As an extension of on-line learning, life-long learning challenges a system which is exposed to patterns from a changing environment during its entire lifespan. An autonomous system should not only integrate new knowledge on-line into its memory, but also preserve the knowledge learned by previous interactions. Thus, life-long learning implies the fundamental Stability-Plasticity Dilemma, which addresses the problem of learning new patterns without forgetting old prototype patterns. We propose an extension to the known Cell Structures, growing Radial Basis Function-like networks, that enables them to learn their number of nodes needed to solve a current task and to dynamically adapt the learning rate of each node separately. As shown in several simulations, the resulting Life-long Learning Cell Structures posses the major characteristics needed to cope with the Stability-Plasticity Dilemma. PMID- 11411638 TI - Pattern classification by a condensed neural network. AB - Neural networks have come to the fore as potent pattern classifiers. More amenable to parallel computation, they are much faster than the nearest neighbor classifier (NN), which, however, has distinctly outperformed them in several applications. The purpose of this study is to investigate a condensed neural network that combines the classification speed of neural networks and the low error rate of the nearest neighbor classifier. This condensed network is a fast, accurate classifier of simple architecture and function: it consists of a set of generalized perceptrons that draw maximal hyperspherical boundaries centered on patterns of memory units, each circumscribing reference patterns of a single category. The generalized perceptrons carry out classification, assisted by sporadic nearest neighbor matching to patterns of a small reference set. We compare the condensed network to a high performance neural network pattern classifier (Kohonen) and to NN in experiments on hand-printed character recognition. PMID- 11411639 TI - A herpes-like virus infects a non-ostreid bivalve species: virus replication in Ruditapes philippinarum larvae. AB - Sporadic high mortalities were reported in June 1997 among hatchery-reared larval Manila clam Ruditapes philippinarum in a French commercial hatchery. Cellular abnormalities were observed in semi-thin sections in affected animals. Transmission electron microscopy revealed the presence of herpes-like virus particles in larvae. This is the first description of a herpes-like virus infection in larval R. philippinarum, a non-ostreid bivalve species. Virus particles were similar to other herpes-like viruses described from different oyster species with respect to ultrastructure and morphogenesis. Electron microscopic examination also demonstrated cells with condensed chromatin and extensive perinuclear fragmentation of chromatin. Like viruses infecting oysters, the herpes-like virus detected in clams may induce apoptosis in infected animals. PMID- 11411640 TI - Characterisation of ISAV proteins from cell culture. AB - The characterisation of 2 infectious salmon anemia virus (ISAV) proteins is described. Proteins were harvested from ISAV-infected Chinook salmon embryo (CHSE)-214 cell culture by continuous elution denaturing gel electrophoresis, enabling the harvest of specific molecular weight fractions. Through the use of a polyclonal antiserum to ISAV, it was possible to identify a potentially autolytic major antigen of 72 kDa and a glycosylated protein of approximately 38 kDa which varied in size depending on cell line compatibility. N-terminal amino acid sequencing of the glycosylated proteins suggests that it is encoded by segment 6 of the ISAV genome. Further, sequence analysis of the glycosylated protein account for the variable molecular weight and may explain differences in host cell compatibility. PMID- 11411641 TI - Sea bass Dicentrarchus labrax nervous necrosis virus isolates with distinct pathogenicity to sea bass larvae. AB - Reproduction of nodavirus disease was performed by experimental infection of sea bass eggs during fertilization or at larval stage 4 with 2 genetically distinguishable nodavirus strains (Sb1 and Sb2) isolated from sea bass collected along the Atlantic and Mediterranean French coast. The pathogenicity of the virus strains was assigned after detection of the virus by ELISA and immunohistochemistry (IHC). The Atlantic (Sb1) strain was more pathogenic than the Mediterranean (Sb2) strain during the fertilization step whilst both strains were pathogenic following experimental exposure of 4 d old larvae. Virus lesions developed in the brain 4 to 6 d following experimental exposure. Experimental ELISA proved very sensitive for detecting the nodavirus in Sb1 or Sb2 experimentally infected larvae, as well as in naturally infected sea bass larvae collected in French hatcheries or in barramundi larvae reared in the Pacific area. The development of an ELISA specific for the 2 nodavirus strains isolated from the sea bass should be useful for the detection of the virus, in addition to other techniques recommended by the Office International des Epizooties (OIE). PMID- 11411642 TI - Immune response to a recombinant capsid protein of striped jack nervous necrosis virus (SJNNV) in turbot Scophthalmus maximus and Atlantic halibut Hippoglossus hippoglossus, and evaluation of a vaccine against SJNNV. AB - Immunisation by intraperitoneal injection of an oil-emulgated recombinant partial capsid protein (rT2) from striped jack nervous necrosis virus (SJNNV) was performed on adult turbot Scophthalmus maximus and Atlantic halibut Hippoglossus hippoglossus. A specific humoral immune response was recorded in both species, and the levels of rT2-specific antibodies increased markedly in all groups during the 20 wk experiment. A challenge model for SJNNV was established by intramuscular injection of juvenile turbot. The turbot developed viral encephalopathy and retinopathy (VER), also known as viral nervous necrosis (VNN), with cumulative mortality in the range of 25 to 66%, after intramuscular inoculation with SJNNV propagated in the striped snake head cell line (SSN-1). Although neither clinical signs nor mortality were registered, SJNNV was neuroinvasive after bath exposure. The infection after both modes of challenge was verified by means of immunohistochemistry and RT-PCR, and SJNNV was reisolated in cell culture. The results indicate that SJNNV may have entered the central nervous system (CNS) by axonal transport through motor nerves after intramuscular inoculation. A vaccine efficacy test was performed on juvenile turbot, employing oil emulsified rT2 as a test vaccine and intramuscular inoculation of SJNNV. Significant protection was observed when the challenge was performed 10 wk post-vaccination. PMID- 11411643 TI - Lactococcus garvieae infection in the giant freshwater prawn Macrobranchium rosenbergii confirmed by polymerase chain reaction and 16S rDNA sequencing. AB - An epizootic bacterial infection in the giant freshwater prawn Macrobranchium rosenbergii occurred in Taiwan from May to June 1999. The cumulative mortality was approximately 30 to 75%. The diseased prawns showed opaque and whitish muscles and were approximately 2 mo old with total lengths from 5 to 6 cm. Histopathologically, they showed marked edema and necrotic lesions with inflammation in the muscles and hepatopancreas. Bacteria isolated using brain heart infusion medium or tryptic soy agar were Gram-positive and ovoid. Three isolates from diseased prawns at different farms were tested using the API 20 Strepsystem and conventional tests and identified as Lactococcus garvieae. Experimental infections with these isolates gave gross signs and histopathological changes similar to those seen in the naturally infected prawns. The LD50 value of isolate MR1 was 6.6 x 10(5) colony forming units/prawn. Identification of MR1 was confirmed by a PCR assay for L. garvieae that gave the expected amplicon of 1100 bp. In addition, its 16S rDNA sequence (GenBank accession number AF283499) gave 99% sequence identity to Enterococcus seriolicida (synonym L. garvieae; GenBank accession number AF061005). This is the first report of confirmed L. garvieae infection in prawn aquaculture. PMID- 11411644 TI - Characterisation of Saprolegnia sp. isolates from channel catfish. AB - Nineteen channel catfish isolates of Saprolegnia sp. obtained from 5 separate fish farms in Mississippi, which became affected by winter kill syndrome during 1991 and 1996, were investigated with respect to physiological characteristics and genetic variation. Isolates of S. parasitica from crayfish and S. diclina were included for comparison. Most strains of catfish isolates grew well at 20 and 30 degrees C. Repeated zoospore emergence was found in catfish isolates of Saprolegnia sp. and S. parasitica, but not in S. diclina. Random amplification of polymorphic DNA polymerase chain reaction (RAPD-PCR) was applied for a further characterisation of the isolates. The RAPD analysis among Saprolegnia spp. isolates was constructed from 686 amplified products in 67 separable positions and indicated that the catfish isolates of Saprolegnia sp. are composed of 3 genetically distinct groups. PMID- 11411645 TI - Induction of proliferative kidney disease (PKD) in rainbow trout Oncorhynchus mykiss via the bryozoan Fredericella sultana infected with Tetracapsula bryosalmonae. AB - Proliferative kidney disease (PKD) is a serious infection of wild and farmed salmonids, affecting mainly the kidney and spleen but becoming systemic in most susceptible fish hosts. This report deals with the transmission of Tetracapsula bryosalmonae Canning, Curry, Feist, Longshaw & Okamura 1999 from naturally infected bryozoans Fredericella sultana Blumenbach 1779 to naive rainbow trout Oncorhynchus mykiss Walbaum 1792, thereby confirming the recent conclusion based on partial 18S rDNA sequence data that bryozoans are hosts of the myxozoan parasite T. bryosalmonae (formerly PKX organism) that causes the disease. Parasite transmission using T. bryosalmonae spores was successful by short-term exposure to disrupted bryozoans known to contain T. bryosalmonae spores and T bryosalmonae sacs liberated from the bryozoans, and by long-term cohabitation with infected bryozoan colonies. Infection was confirmed by examination of kidney imprints, detection of the parasite in stained tissue sections, PCR using T. bryosalmonae-specific primers, and comparison of amplified 18S rDNA sequences from the bryozoans and experimentally infected fish. Transmission was not apparent, nor was PKD induced, in fish challenged by intraperitoneal injection of spores isolated from F. sultana. PMID- 11411646 TI - Long-term monitoring on the occurrence of a myxosporean parasite Kudoa camarguensis (Myxosporean) on the common goby (Teleostei, pisces) Pomatoschistus microps. AB - The evolution of a host-parasite system composed of Pomatoschistus microps-Kudoa camarguensis was investigated in the Vaccares lagoon (Rhjne river Delta, France) from 1993 to 1997. During this long-term monitoring, centennial flooding of the Rhjne river occurred, leading to an inrush of about 110 million m3 of freshwater in the Vaccares lagoon. The salinity drastically dropped from 14 to 5 g l(-1) in 1 wk. We observed that the annual prevalence and abundance of the myxosporean parasite decreased from 12.18 in 1993 to 3.7% in 1997 and from 1.10 in 1993 to 0.27 in 1997, respectively. Here, we discuss the possible reasons for the rapid decline of this host-parasite system following the flood. PMID- 11411647 TI - Effects of bacteria on the growth of an amoeba infecting the gills of turbot. AB - We analysed the influence of various bacteria on the in vitro growth of trophozoites of a Platyamoeba strain isolated from diseased gill tissues of cultured turbot. Little or no growth was shown by amoebae cultured in the presence of (1) the turbot-pathogenic bacteria Vibrio anguillarum, Aeromonas salmonicida or Streptococcus sp., (2) Pasteurella piscicida or Vibrio vulnificus (pathogenic for some fishes but not turbot), or (3) the non-pathogenic 'environmental' bacteria Vibrio campbelli, Vibrio fluvialis or Pseudomonas dondorofii. The only bacteria which were successfully utilized as food sources were Aeromonas hydrophila (pathogenic for some fishes but not turbot) and the non pathogens Vibrio natriegens, Pseudomonas nautica and Escherichia coli. These results suggest that the colonization of the gills of cultured turbot by the epizoic amoeba Platyamoeba may be an indicator of faecal contamination. PMID- 11411648 TI - Hyperventilation and loss of hemolymph Na+ and Cl- in the freshwater amphipod Gammarus fossarum exposed to acid stress: a preliminary study. AB - The effect of acidification on the acid-sensitive species Gammarus fossarum was investigated in the laboratory. The results showed that as mortality increased, mean hemolymph chloride and sodium concentrations decreased rapidly. Concomitantly, organisms hyperventilated during the first 24 h and then started to hypoventilate. These results demonstrated that exposure to acid stress in the acid-sensitive species G. fossarum led to ion-regulatory and respiratory failure as previously reported in fish and crayfish exposed to acid stress. PMID- 11411649 TI - Isolation and identification of infectious salmon anaemia virus (ISAV) from Coho salmon in Chile. AB - The isolation of infectious salmon anaemia virus (ISAV) from asymptomatic wild fish species including wild salmon, sea trout and eel established that wild fish can be a reservoir of ISAV for farmed Atlantic salmon. This report characterizes the biological properties of ISAV isolated from a disease outbreak in farmed Coho salmon in Chile and compares it with ISAV isolated from farmed Atlantic salmon in Canada and Europe. The virus that was isolated from Coho salmon tissues was initially detected with ISAV-specific RT-PCR (reverse transcription-polymerase chain reaction). The ability of the virus to grow in cell culture was poor, as cytopathology was not always conspicuous and isolation required passage in the presence of trypsin. Virus replication in cell culture was detected by RT-PCR and IFAT (indirect fluorescent antibody test), and the virus morphology was confirmed by positive staining electron microscopy. Further analysis of the Chilean virus revealed similarities to Canadian ISAV isolates in their ability to grow in the CHSE-214 cell line and in viral protein profile. Sequence analysis of genome segment 2, which encodes the viral RNA polymerase PB1, and segment 8, which encodes the nonstructural proteins NS1 and NS2, showed the Chilean virus to be very similar to Canadian strains of ISAV. This high sequence similarity of ISAV strains of geographically distinct origins illustrates the highly conserved nature of ISAV proteins PB1, NS1 and NS2 of ISAV. It is noteworthy that ISAV was associated with disease outbreaks in farmed Coho salmon in Chile without corresponding clinical disease in farmed Atlantic salmon. This outbreak, which produced high mortality in Coho salmon due to ISAV, is unique and may represent the introduction of the virus to a native wild fish population or a new strain of ISAV. PMID- 11411650 TI - The nested variant of transitional cell carcinoma--a rare neoplasm with poor prognosis. AB - The "nested" variant of transitional cell carcinoma is a rare variant composed of apparently clusters of urothelial cells, of benign appearance, which often simulate von Brunn's nests and invade into the lamina propria or deeper. Only 36 cases have been reported to date and herein we report an additional 10 cases. Of these 10 cases treated with locoregional therapy, seven died of disease or treatment complications 4-40 months after diagnosis while one patient died of unrelated disease after 90 months. Follow-up is < or = 1 year for the remaining two patients. Our data and a review of the literature supports the assertion that the nested variant of transitional cell carcinoma is an aggressive neoplasm. PMID- 11411651 TI - The importance of glucose for the Escherichia coli mediated citrate depletion in synthetic and human urine. AB - OBJECTIVE: Escherichia coli has previously been shown to reduce urine citrate and influence urine pH. In this study the influence of glucose on the E. coli mediated citrate decrease has been investigated. MATERIAL AND METHODS: In synthetic urine, a glucose concentration of > or = 1 mmol/l was a prerequisite for bacteria to grow and lower citrate. At glucose concentrations > or = 5 mmol/l an E. coli mediated pH decrease correlated to urine glucose was observed. RESULTS: In human urine, variations in urine glucose influenced the citrate decrease and addition of glucose accelerated the E. coli mediated citrate decrease, which in certain urines could be very pronounced. CONCLUSIONS: Citrate has a pronounced effect on the activity product of calcium oxalate and calcium phosphate and the E. coli mediated decrease in urine citrate may be involved in the formation of urinary tract stones and catheter encrustations. PMID- 11411652 TI - Urinary tract infection in children associated with idiopathic hypercalciuria. AB - OBJECTIVE: The aim of this study was to evaluate the association of symptomatic non-calculous idiopathic hypercalciuria (IH) with urinary tract infection (UTI) in children. MATERIAL AND METHODS: This was a retrospective case review of children who had urinary calcium excretion greater than 2 mg/kg/day or random urine calcium-creatinine ratio (UCa/UCr) greater than 0.18 mg/mg. RESULTS: One hundred and twenty-four consecutive children with clinical complaints and elevated urine calcium excretion were reviewed. Fifty children (40%) had UTI of which 39 (78%) had recurrent UTI. There was no difference in age between children with UTI and those without UTI. Twenty-four-hour urine calcium and random UCa/UCr were also not different. Only 4 children (8%) had renal stones whereas hematuria, abdominal pain and urine incontinence were frequent associated findings. Six of the children with recurrent UTI (15%) had an anatomical urinary tract abnormality. Therapy in all children consisted of increased fluid intake and reduction in diet sodium and oxalate; however, 14 of the 39 children with recurrent UTI (36%) required therapy with a thiazide diuretic. Recurrent UTI was abolished in 24 children, one child had a single recurrence and 4 children had no response to treatment. CONCLUSIONS: We propose that non-calculous IH may be an important contributing factor to recurrent UTI in children. PMID- 11411653 TI - Smokeless nicotinergic stimulation of vasopressin secretion in patients with persisting nocturnal enuresis and controls. AB - OBJECTIVE: This study aimed to evaluate the effect of nicotine stimulation on pituitary release of plasma arginine vasopressin (P(AVP)) in patients with primary monosymptomatic nocturnal enuresis (PMNE) and healthy control subjects. MATERIAL AND METHODS: Postpubertal teenagers and adult enuretics as well as control subjects were enrolled into the study and admitted to hospital for measurements of P(AVP) in relation to intake of orally administered nicotine. Sixteen patients with PMNE (9 females, 7 males; aged 15-51 years, mean 23.5) and nine normal subjects (4 females, 5 males; aged 24-31 years, mean 27.3) were studied. The enuretics were characterized prior to investigation as either 1 desamino-8-D-arginine vasopressin (DDAVP) responders (n = 8; 16-51 years, mean 28.9) or DDAVP non-responders (n = 8; 15-24 years, mean 18.1) based on the reduction in the number of wet nights. P(AVP), mean arterial blood pressure (MAP) and heart rate (HR) were measured 0, 5, 10, 15 and 30 min, and plasma osmolality (P(osm)) 0 and 30 min after receiving 4 mg nicotine (Nicorette, Pharmacia & Upjohn) chewing gum. RESULTS: In the compiled material a slight but statistically significant increase was observed in P(AVP) at 30 min compared with baseline levels, concurrent with significant rises in MAP and HR above baseline levels at 15 and 30 min. No difference was seen in P(osm) before and after nicotine administration. No other significant variation over time, assessed by an ANOVA, was detected. No difference was encountered in any measured parameter between controls and enuretics or between DDAVP responders and non-responders. CONCLUSION: Smokeless nicotine chewing gum induces non-osmotic vasopressin release in humans. The secretory AVP capacity to this stimulation is normal in PMNE. PMID- 11411654 TI - Circulating insulin-like growth factor-I and benign prostatic hyperplasia--a prospective study. AB - OBJECTIVE: The aim of this study was to investigate the role of insulin-like growth factor-I (IGF-I), a strongly mitogenic and anti-apoptotic factor, in the development of benign prostatic hyperplasia (BPH). The bioactivity of IGF-I within tissues depends on circulating levels, as well as on the local production of IGF-I and the presence of IGF-binding proteins (IGFBPs). The IGFBPs regulate the efflux of IGF-I to the extravascular space and the bioavailability of IGF-I within tissues. MATERIAL AND METHODS: Within the Northern Sweden Health and Disease Study, 60 cases of BPH defined by a history of prostate resection were identified, and two controls per case were selected. IGF-I, IGFBP-1, IGFBP-3 and insulin were measured by immuno-radiometric assays in stored plasma samples drawn a mean of 3.2 years before surgery. RESULTS: The risk of BPH increased with increasing quartile levels of IGF-I adjusted for IGFBP-3 (p(trend) = 0.10) up to a relative risk of 2.16 (95% confidence interval 0.83-5.64) for the highest quartile. The risk decreased with increasing levels of IGFBP-1 (p(trend) = 0.10). CONCLUSIONS: Our results suggest that elevated IGF-I bioactivity may stimulate the development of BPH; however, they were not statistically significant and require confirmation from larger studies. PMID- 11411655 TI - The effect of transurethral needle ablation on symptoms of chronic pelvic pain syndrome--a pilot study. AB - OBJECTIVE: We evaluated the effects of transurethral needle ablation (TUNA) in chronic pelvic pain syndrome (CPPS). MATERIAL AND METHODS: CPPS patients were subjected to TUNA (n = 21) or sham therapy (urethrocystoscopy) (n = 6). Digital rectal examination and transrectal ultrasound were performed. Prostate-specific antigen and urine were analysed before and 1 month after treatment. Symptoms, the DAN-PSS-1 questionnaire, urine flow, residual urine volume, medication and a subjective estimation of the effect of treatment were evaluated during a follow up period of 12 months. RESULTS: In both groups the maximal urine flow increased and the DAN-PSS-1 score decreased, but the decrease in the DAN-PSS-1 score was significant (p = 0.002) only in the TUNA group. The total symptom score decreased from 14 to 8.3 points in the TUNA group, and from 12.6 to 8.0 points in sham patients. TUNA relieved pain significantly. In the TUNA group symptoms were ameliorated in patients with small (< or = 20 ml; p = 0.002) and large prostates (> 20 ml; p = 0.04). Similarly, patients with both mild (p = 0.004) and severe (p = 0.02) symptoms at baseline benefitted from TUNA. Of the TUNA patients, 72% felt better at 1-year follow-up, although 57% still had symptoms and 38% needed medication. The figures in the sham group were 50%, 66% and 50%, respectively. CONCLUSIONS: TUNA relieved symptoms in CPPS patients for at least 12 months. The need for medication and the presence of symptoms were reduced, and the fraction of patients satisfied with treatment was higher in the TUNA group than in the sham group. However, these differences between the groups were not statistically significant. To assess the clinical value of TUNA in CPPS a prospective double blind study with a large number of patients is needed to confirm the results of this pilot study. PMID- 11411656 TI - Vasovasostomy--a 15-year experience. AB - OBJECTIVE: This study aimed to evaluate the results of vasovasostomies performed in the authors' clinic over 15 years. MATERIAL AND METHODS: Between 1983 and 1998 39 vasovasostomies were performed using a macroscopic technique. The medical charts were reviewed and a postal survey was sent to 36 of these patients. The patients were invited to visit the outpatient department and to give semen for analysis and a blood sample for determination of serum antisperm antibodies and serum follicle-stimulating hormone. Twenty-five patients filled in the questionnaire and 10 patients were obtained for further semen and serum analyses. RESULTS: The overall pregnancy rate was 56% (14/25 respondents). Three of these 14 men used an assisted reproduction method with their own ejaculated sperm. The postoperative patency was recorded in the medical charts only sporadically. The age and the interval between the vasectomy and vasectomy reversal averaged 42 years and 8.6 years, respectively. Those men who could father a child had a somewhat shorter mean obstructive interval than those who failed (8.0 vs 9.3 years), although the difference was not statistically significant in this small patient sample. The serum follicle-stimulating hormone concentrations were normal, with no association with the pregnancy rate. CONCLUSION: Macroscopic vasovasostomy is an effective means of re-establishing fertility in vasectomized men. PMID- 11411657 TI - Influence of erythropoietin treatment on gonadotropic hormone levels and sexual function in male uremic patients. AB - OBJECTIVE: The purpose of this study was to evaluate whether levels of sex hormones and sexual function differ in renal failure patients with and without uremia and the effect of treatment with recombinant human erythropoietin (rhuEPO). MATERIAL AND METHODS: Fifteen males with chronic renal failure who were not receiving hemodialysis and 25 male renal failure patients with uremia who were undergoing hemodialysis were enrolled before and after rhuEPO therapy. Fifteen male volunteers matched for age and weight were also studied. Levels of various blood biochemicals were measured in all patients before and 1 week after rhuEPO treatment. Sexual function was also studied in all patients before and 6 months after rhuEPO treatment. RESULTS: The control group had significantly higher levels of testosterone (6.21 +/- 1.21 ng/ml) and hematocrit (Hct) (43.2 +/ 2.1%) and significantly lower levels of prolactin (5.27 +/- 1.21 ng/ml), follicular-stimulating hormone (FSH) (7.51 +/- 2.36 mIU/ml) and leutinizing hormone (LH) (4.23 +/- 2.10 mIU/ml) than the two patient groups (p < 0.05 for all comparisons). Patients with renal failure only had significantly lower levels of testosterone and Hct (2.54 +/- 0.53 ng/ml and 21.4 +/- 1.4%, respectively) than those with uremia (3.65 +/- 0.52 ng/ml and 24.3 +/- 2.5%, respectively; p < 0.001 for both comparisons). After rhuEPO therapy, the testosterone and Hct levels of the two patient groups did not reach the level of the control subjects (p < 0.05 for both comparisons). Similarly, the levels of prolactin, FSH and LH were significantly higher in both patient groups than those of control subjects after rhuEPO therapy (p < 0.001 for both comparisons). However, after rhuEPO therapy, significant increases in testosterone and Hct levels were found in both patient groups (p < 0.001 for both comparisons). Sexual function was also markedly improved in the hemodialysis patient group. While 20/25 (80%) male hemodialysis patients reported improved sexual function after rhuEPO treatment, only 3/15 (20%) chronic renal failure patients reported improvement. CONCLUSIONS: In patients with advanced uremia, rhuEPO therapy may result in improved gonadotropic hormone levels and sexual function. Good dialysis quality may contribute to the increase in the incidence of patients with better sexual function. PMID- 11411658 TI - Mechanism of increased serum soluble interleukin-2 receptor levels in patients on continuous ambulatory peritoneal dialysis. AB - OBJECTIVE: The aim of this study was to investigate the mechanism underlying the increase of serum soluble interleukin-2 receptor (IL-2R) levels in patients on continuous ambulatory peritoneal dialysis. MATERIAL AND METHODS: In 13 dialysis patients and 17 healthy controls, serum soluble IL-2R levels were determined by enzyme-linked immunosorbent assay, and CD25-positive (cell surface IL-2R positive) cells were detected by flow cytometry. Soluble IL-2R levels were also measured in the supernatant of cultured peripheral blood mononuclear cells. RESULTS: The serum soluble IL-2R level was significantly higher in the patients than in the healthy controls (p < 0.0001). In contrast, both the percentage and the absolute count of CD25-positive cells showed no significant differences, and neither did the soluble IL-2R level in culture supernatant. Serum soluble IL-2R levels showed a positive correlation with the serum beta2-microglobulin level (p < 0.01), the age of the patients (p < 0.05), and duration of dialysis (p < 0.05), as well as a negative correlation with the urine volume (p < 0.05). CONCLUSIONS: The increase of serum soluble IL-2R in patients on peritoneal dialysis may be caused by accumulation due to its low transperitoneal clearance and low urinary excretion. PMID- 11411659 TI - Retroperitoneal cystic papillary carcinoma of probable nephrogenic origin. AB - A case of primary retroperitoneal cystic papillary carcinoma of probable nephrogenic origin is presented. To our knowledge, this is the first such case reported in the literature. PMID- 11411660 TI - Bilateral multifocal renal oncocytoma--a case report and literature review. AB - A case of bilateral multifocal renal oncocytoma is reported. The incidence, diagnosis and management of such tumors is discussed. PMID- 11411661 TI - A rare cause of cystic testicular mass in an infant--cystic dysplasia of the testis. AB - In this article, a 6-month-old infant with cystic dysplasia of the testis (CDT) is presented. Although different genitourinary abnormalities associated with CDT have been reported, this is the first case of the association of CDT with penoscrotal hypospadias, unilateral undescended testicle, bilateral scrotal hernia, small and hypocompliant bladder and urethral obstruction. Orchiectomy was carried out for the treatment of this rare cause of testicular cystic mass. PMID- 11411662 TI - Antiglomerular basement membrane antibody-crescentic glomerulonephritis complicating chronic bronchiectasis. AB - A 68-year-old woman with chronic bronchiectasis presented with haematuria and severe oligoanuric renal failure with no other serious systemic manifestation. Antiglomerular basement membrane (anti-GBM) antibodies and anti-myeloperoxidase antibodies were positive. Renal biopsy revealed anti-GBM crescentic glomerulonephritis. A conservative approach was followed and the patient is stable on chronic haemodialysis 6 months later. To the authors' knowledge, there has only been one previous report of anti-GBM disease complicating bronchiectasis. PMID- 11411663 TI - Odynorgasmia. AB - Painful ejaculation ("odynorgasmia") is not well recognized. When it occurs it may indicate the precise site of pathology. PMID- 11411664 TI - Effect of thromboxane and nitric oxide blockade on renal blood flow increase during volume expansion in hydronephrotic rats. AB - OBJECTIVE: It has recently been found that hydronephrotic rats, despite low diuresis, show a significant increase in renal blood flow (RBF) during volume expansion. The present experiments were designed to evaluate the mechanisms underlying this phenomenon. MATERIAL AND METHODS: Three-week-old Sprague-Dawley rats underwent partial obstruction of the left ureter using the Ulm-Miller psoas groove technique. The effects on RBF were studied 3 weeks later under general anesthesia using real-time ultrasound flowmetry, first during normohydration and then during extracellular volume expansion, in both untreated animals, and after prior blockade of either thromboxane or nitric oxide. RESULTS: Significant hydronephrosis developed in all cases. RBF was normal under control conditions. During volume expansion RBF increased significantly in untreated experimental animals (mean 7.5%). In contrast to this finding, RBF remained unchanged during volume expansion in both the thromboxane and nitric oxide blockade groups. CONCLUSION: It is concluded that a thromboxane- and/or nitric oxide-dependent RBF redistribution takes place in hydronephrotic kidneys during volume expansion. PMID- 11411665 TI - Urinary excretion of calcitonin gene-related peptide in males with hot flushes after castration for carcinoma of the prostate. AB - OBJECTIVE: The majority of men who undergo surgical or medical castration due to prostatic carcinoma develop vasomotor symptoms with hot flushes. The mechanisms behind these symptoms are poorly understood. One possible explanation is a release of the vasodilatory peptide calcitonin gene-related peptide (CGRP) from perivascular nerves, which seem to be involved in the mechanisms behind vasomotion and sweating in postmenopausal women. The aim of this report was to investigate whether CGRP is involved in vasomotion in men after castration therapy. MATERIAL AND METHODS: Twenty-four hour urine excretion of CGRP was analysed in 15 men with prostatic carcinoma, using radioimmunoassay before and 3 months after surgical or medical castration. RESULTS: Eleven of the 15 men developed hot flushes during the observation period of 3 months. Twenty-four hour urine excretion of CGRP did not change significantly after castration, either in the group as a whole or in those 11 men who developed hot flushes. CONCLUSIONS: Even though we did not observe any significant changes in 24-h urine excretion of the potent vasodilator CGRP after castration it is possible that serum levels of CGRP increase during hot flushes, without having an effect on the 24-h urine excretion of the peptide. PMID- 11411666 TI - Emotional isolation: prevalence and the effect on well-being among 50-80-year-old prostate cancer patients. AB - OBJECTIVE: To investigate to what extent prostate cancer patients confide their emotional concerns, and whether having no one to confide in affects well-being. MATERIAL AND METHODS: A population-based study using epidemiological methods. A questionnaire was mailed to all 431 living prostate cancer patients aged 50-80 at the time of selection, diagnosed 1.5-2 years previously in Stockholm County, and 435 randomly selected men in the same age group. The questionnaire was completed anonymously. The main outcome measures included questions assessing the extent to which the men could share emotionally taxing feelings with their partner or others and questions assessing well-being. RESULTS: The questionnaire was returned by 79% of the patients and by 73% of the randomly selected men. Approximately one in five patients had no one to confide in. Of patients living with a partner, only one in 10 confided in someone other than their partner. Three out of 10 patients living in a relationship could not confide in their partner. Men having no one to confide in were less content with their life and reported poorer psychological and overall well-being compared with other men. The prostate cancer patients were not more likely to have someone to confide in than men in general. CONCLUSIONS: The results indicate that a lack of emotional support may be a problem for many prostate cancer patients and that the traditional psychosocial support offered to most cancer patients in Sweden may not reach male patients. There may be a need for a gender-adapted approach to emotional support. PMID- 11411667 TI - Reclassification of Cellulomonas cellulans (Stackebrandt and Keddie 1986) as Cellulosimicrobium cellulans gen. nov., comb. nov. AB - Phylogenetic analysis of 16S rDNA provides evidence that Cellulomonas cellulans branches outside the phylogenetic confines of the genus Cellulomonas. The distinct phylogenetic and chemotaxonomic status of Cellulomonas cellulans as a phylogenetic neighbour of the genus Promicromonospora, justifies the description of a new genus for which the name Cellulosimicrobium gen. nov. with the type species Cellulosimicrobium cellulans comb. nov. is proposed. PMID- 11411668 TI - Mesorhizobium chacoense sp. nov., a novel species that nodulates Prosopis alba in the Chaco Arido region (Argentina). AB - Low-molecular-weight RNA analysis was performed for the identification and classification of 20 Argentinian strains isolated from the root nodules of Prosopis alba. SDS-PAGE of total cellular proteins, determination of the DNA base composition, DNA-DNA reassociation experiments and physiological and biochemical tests were also carried out for these strains and the whole 16S rRNA gene was sequenced from one of the strains, strain LMG 19008T. Results of the genotypic and phenotypic characterization showed that the strains isolated in this study belong to a group that clustered in the genus Mesorhizobium. The results of DNA DNA hybridizations showed that this group is a novel species of this genus. The name Mesorhizobium chacoense sp. nov. is proposed for this species. The type strain is LMG 19008T (= CECT 5336T). PMID- 11411669 TI - Agrobacterium larrymoorei sp. nov., a pathogen isolated from aerial tumours of Ficus benjamina. AB - Tumorigenic Agrobacterium strains isolated from tumours growing on pruned branches of Ficus benjamina have previously been shown to have unique opine metabolism and sufficient 16S rRNA sequence differences to suggest that they belong to a new species. DNA-DNA hybridization results confirmed that these strains represent a new species and Agrobacterium larrymoorei sp. nov. (type strain ATCC 51759T = CFBP 5473T = NCPPB 4096T) is proposed as the name for the species. PMID- 11411670 TI - Shewanella japonica sp. nov. AB - Two strains of agar-digesting bacteria, KMM 3299T and KMM 3300, respectively isolated from sea water and the mussel Protothaca jedoensis, have been characterized. Based on sequencing of the 16S rRNA gene, KMM 3299T showed the highest similarity (93-95%) to members of the genus Shewanella. The G+C contents of the DNAs of these strains were 43-44 mol%. The level of DNA homology between the two strains was conspecific (95%), indicating that they represent a distinct genospecies. These organisms were non-pigmented, Gram-negative, polarly flagellated, facultatively anaerobic, mesophilic, neutrophilic and able to degrade a wide range of high molecular mass polymers, including alginate, carrageenan, laminaran and agar. The novel organisms were susceptible to gentamycin, carbenicillin, lincomycin and oleandomycin. The predominant cellular fatty acids were i-15:0, 16:0, 16:1(n-7), 18:1(n-7). Eicosapentaenoic acid, 20:5(n-3), was detected in the two isolates at levels of 1-8%, depending on the temperature of cultivation. Phylogenetic evidence, together with phenotypic characteristics, showed that the two isolates studied constitute a novel species of the genus Shewanella. The name Shewanella japonica is proposed; the type strain is KMM 3299T(= LMG 19691T = CIP 106860T). PMID- 11411671 TI - Isolation of Desulfomicrobium orale sp. nov. and Desulfovibrio strain NY682, oral sulfate-reducing bacteria involved in human periodontal disease. AB - The species of sulfate-reducing bacteria that prevail in sites affected by periodontal disease may be different from those commonly occurring in the digestive tracts of healthy individuals. Ten strains of mesophilic sulfate reducing bacteria (SRB) were isolated from subgingival plaque in periodontal lesions of ten patients with periodontitis. Characterization on the basis of morphological, physiological and phylogenetic properties demonstrated two distinct types of oral SRB. One strain was a curved rod with high motility. For dissimilatory sulfate reduction, lactate or pyruvate was oxidized incompletely to equimolar amounts of acetate. Desulfoviridin and cytochrome c3 were present in this mesophilic vibrio and the cellular lipid profile was similar to that from members of the genus Desulfovibrio. The 16S rDNA sequence was similar to that of the proposed 'Desulfovibrio fairfieldensis'. Cells of the nine other strains were straight, rod-shaped, exhibited a low growth rate and oxidized substrates incompletely to acetate. These SRB, like members of the genus Desulfomicrobium, lacked desulfoviridin. Analysis of the 16S rDNA sequences of seven of the nine isolates showed a high degree of similarity among these oral strains, forming a distinct lineage within the genus Desulfomicrobium. The cellular lipid profile of a representative oral strain, NY678T, was in accordance with that of other Desulfomicrobium species, but also showed dissimilar features. The phenotypic and phylogenetic analyses indicate that these rod-shaped SRB from the oral cavity could be regarded as a new species, for which the designation Desulfomicrobium orale sp. nov. is proposed. PMID- 11411672 TI - Revival of the genus Lentzea and proposal for Lechevalieria gen. nov. AB - The genus Saccharothrix is phylogenetically heterogeneous on the basis of analysis of almost complete 16S rDNA sequences. An evaluation of chemotaxonomic, morphological and physiological properties in the light of the molecular phylogeny data revealed that several species are misclassified. Saccharothrix aerocolonigenes NRRL B-3298T and Saccharothrix flava NRRL B-16131T constitute a lineage distinct from Saccharothrix and separate from Lentzea. The genus Lechevalieria gen. nov. is proposed for these species. Lechevalieria aerocolonigenes comb. nov. is the type species and S. flava is transferred as Lechevalieria flava comb. nov. Although Lentzea albidocapillata, the type species of the genus Lentzea, was transferred recently to the genus Saccharothrix, the revival of Lentzea is clearly supported by molecular phylogenetic and chemotaxonomic data. The description of the revived genus is emended to include galactose, mannose and traces of ribose as diagnostic whole-cell sugars and MK 9(H4) as the principal menaquinone and elimination of tuberculostearic acid as a diagnostic component in the fatty acid profile. Saccharothrix waywayandensis NRRL B-16159T, S. aerocolonigenes NRRL B-16137 and 'Asiosporangium albidum' IFO 16102 are members of the amended genus Lentzea on the basis of phylogenetic and chemotaxonomic properties. S. waywayandensis is transferred to Lentzea as Lentzea waywayandensis comb. nov., while the new species Lentzea californiensis sp. nov. and Lentzea albida sp. nov. are described for S. aerocolonigenes NRRL B-16137 and 'A. albidum' IFO 16102, respectively. Nucleotide signatures in the 16S rDNA sequences are defined that are diagnostic for the genera Lechevalieria, Lentzea and Saccharothrix. PMID- 11411673 TI - Albibacter methylovorans gen. nov., sp. nov., a novel aerobic, facultatively autotrophic and methylotrophic bacterium that utilizes dichloromethane. AB - A novel genus, Albibacter, with one species, Albibacter methylovorans sp. nov., is proposed for a facultatively chemolithotrophic and methylotrophic bacterium (strain DM10T) with the ribulose bisphosphate (RuBP) pathway of C1 assimilation. The bacterium is a Gram-negative, aerobic, asporogenous, nonmotile, colourless rod that multiplies by binary fission. The organism utilizes dichloromethane, methanol, methylamine, formate and CO2/H2, as well as a variety of polycarbon compounds, as carbon and energy sources. It is neutrophilic and mesophilic. The major cellular fatty acids are straight-chain unsaturated C18:1, saturated C16:0 and cyclopropane C19:0 acids. The main ubiquinone is Q-10. The dominant phospholipids are phosphatidyl ethanolamine, phosphatidyl glycerol, phosphatidyl choline and cardiolipin. The DNA G+C content is 66.7 mol%. Strain DM10T has a very low degree of DNA-DNA hybridization (4-7%) with the type species of the genera Paracoccus, Xanthobacter, Blastobacter, Angulomicrobium, Ancylobacter and Ralstonia of RuBP pathway methylobacteria. Another approach, involving comparative 16S rDNA analysis, has shown that the novel isolate represents a separate branch within the alpha-2 subgroup of the Proteobacteria. The type species of the new genus is Albibacter methylovorans sp. nov.; the type strain is DM10T (= VKM B-2236T = DSM 13819T). PMID- 11411674 TI - Taxonomic characterization of Ketogulonigenium vulgare gen. nov., sp. nov. and Ketogulonigenium robustum sp. nov., which oxidize L-sorbose to 2-keto-L-gulonic acid. AB - Four bacterial strains that oxidize L-sorbose to 2-keto-L-gulonic acid, a key intermediate in the synthesis of vitamin C, were isolated from soils of geographically distinct locations. All were Gram-negative, facultatively anaerobic, chemoheterotrophic rods. Comparative analysis revealed nearly identical 16S rDNA sequences amongst them (99.7-100% identical) and identified them as members of the alpha-subclass of the Proteobacteria. Phylogenetic analysis identified the closest taxonomically defined genus as Roseobacter (92.1 92.8% identical). On the basis of phylogenetic, phenotypic and genotypic analyses, a new genus is proposed, Ketogulonigenium gen. nov. Based upon these analyses, we also propose the reclassification of strain DSM 4025TP, originally identified as Gluconobacter oxydans, to the genus Ketogulonigenium. Two species are proposed: the type species Ketogulonigenium vulgare gen. nov., sp. nov., consisting of strains 62A-12APP, 266-13BPP and the type strain K. vulgare DSM 4025TP, and Ketogulonigenium robustum gen. nov., sp. nov., consisting of the type strain K. robustum X6LTP (= NRRL B-21627 = KCTC 0858BP). The species affiliation of the fifth strain (291-19PP) remains unresolved. PMID- 11411675 TI - Retrieval of the species Alteromonas tetraodonis Simidu et al. 1990 as Pseudoalteromonas tetraodonis comb. nov. and emendation of description. AB - A polyphasic taxonomy study was undertaken of three strains of Pseudoalteromonas haloplanktis subsp. tetraodonis (Simidu et al. 1990) Gauthier et al. 1995. DNA was prepared from each of the strains and genomic relatedness was measured by DNA DNA hybridization. Strains KMM 458T and IAM 14160T shared 99% genetic relatedness, but were only 48-49% related to the type strain of Pseudoalteromonas haloplanktis subsp. haloplanktis, IAM 12915T. The third strain, P. haloplanktis subsp. tetraodonis A-M, showed 83% genetic similarity with P. haloplanktis subsp. haloplanktis IAM 12915T and 32% with KMM 458T. From these results, it is concluded that strains KMM 458T and IAM 14160T comprise a separate species, originally described as Alteromonas tetraodonis, whereas strain A-M belongs to the species Pseudoalteromonas haloplanktis. Based on phenotypic and chemotaxonomic data, genomic fingerprint patterns, DNA-DNA hybridization data and phylogenetic analysis of 16S rRNA, it is proposed that the species Alteromonas tetraodonis be retrieved and recognized as Pseudoalteromonas tetraodonis comb. nov. (type strain IAM 14160T = KMM 458T). PMID- 11411676 TI - Sporosarcina aquimarina sp. nov., a bacterium isolated from seawater in Korea, and transfer of Bacillus globisporus (Larkin and Stokes 1967), Bacillus psychrophilus (Nakamura 1984) and Bacillus pasteurii (Chester 1898) to the genus Sporosarcina as Sporosarcina globispora comb. nov., Sporosarcina psychrophila comb. nov. and Sporosarcina pasteurii comb. nov., and emended description of th. AB - A light-orange-coloured, facultatively anaerobic, rod-shaped bacterium (strain SW28T), which was isolated from seawater in Korea, was taxonomically studied by a polyphasic approach. This organism formed round terminal endospores in swollen sporangia. The peptidoglycan type is A4alpha, based on L-Lys-L-Ala-D-Asp. The predominant menaquinone is MK-7 and the major fatty acid is ante-C15:0. The G+C content of the DNA is 40 mol%. Phylogenetic analysis based on 16S rDNA sequences showed that strain SW28T falls within the radiation of a cluster comprising the rRNA group 2 bacilli and non-Bacillus-type organisms. Strain SW28T showed the highest degree of relatedness to the type strain of Sporosarcina ureae, sharing 96.8% 16S rDNA similarity. Levels of DNA-DNA relatedness between strain SW28T and S. ureae DSM 2281T and the type strains of some Bacillus species forming a coherent phylogenetic cluster are less than 12.5%. On the basis of phenotypic and chemotaxonomic characteristics, 16S rDNA sequence analysis and DNA-DNA relatedness data, it is proposed that strain SW28T should be placed in the genus Sporosarcina as a new species, Sporosarcina aquimarina sp. nov. The type strain is SW28T (= KCCM 41039T = JCM 10887T). From the results of the taxonomic re evaluation, it is also proposed that Bacillus globisporus, Bacillus psychrophilus and Bacillus pasteurii be transferred to the genus Sporosarcina as Sporosarcina globispora, Sporosarcina psychrophila and Sporosarcina pasteurii, respectively. PMID- 11411677 TI - Bacillus jeotgali sp. nov., isolated from jeotgal, Korean traditional fermented seafood. AB - Two Gram-variable, rod-shaped, endospore-forming bacterial strains, which are motile with peritrichous flagella, were isolated from a Korean traditional fermented seafood, jeotgal. The two isolates (strains YKJ-1OT and YKJ-11) were proven to be members of the same species on the basis of the results of phenotypic and phylogenetic analyses and DNA relatedness. Strains YKJ-10T and YKJ 11 were characterized by having cell wall peptidoglycan based on meso diaminopimelic acid, MK-7 as the predominant menaquinone, and iso-C15:0 as the major fatty acid. The G+C content of the DNA was 41 mol%. Strains YKJ-10T and YKJ 11 showed only a 1 bp sequence difference in the 16S rDNA sequences. The two strains formed distinct phylogenetic lineages within the radiation of the cluster comprising Bacillus species. Levels of 16S rDNA similarity between strains YKJ 10T and YKJ-11 and Bacillus species were less than 96.6%. Levels of DNA-DNA relatedness were found to be low enough to distinguish strains YKJ-10T and YKJ-11 from some phylogenetically related Bacillus species. On the basis of phenotypic properties, phylogeny and genomic distinctiveness, strains YKJ-10T and YKJ-11 represent a new species of the genus Bacillus, for which a new name, Bacillusjeotgali sp. nov., is proposed. The type strain of the new species is strain YKJ-10T (= KCCM 41040T = JCM 10885T). PMID- 11411678 TI - Burkholderia fungorum sp. nov. and Burkholderia caledonica sp. nov., two new species isolated from the environment, animals and human clinical samples. AB - A polyphasic taxonomic study that included DNA-DNA hybridizations, DNA base ratio determinations, 16S rDNA sequence analyses, whole-cell protein and fatty acid analyses and an extensive biochemical characterization was performed on 16 strains isolated from the environment, animals and human clinical samples. The isolates belonged to the genus Burkholderia, were phylogenetically closely related to Burkholderia graminis, Burkholderia caribensis and Burkholderia phenazinium and had G+C contents between 61.9 and 62.2 mol%. Seven strains isolated from the rhizosphere were assigned to Burkholderia caledonica sp. nov. [type strain LMG 19076T (= CCUG 42236T)]. Nine strains isolated from the environment, animals and human clinical samples were assigned to Burkholderia fungorum sp. nov. [type strain LMG 16225T (= CCUG 31961T)]. Differential tests for B. graminis, B. caribensis, B. phenazinium, B. caledonica and B. fungorum include the following: assimilation of trehalose, citrate, DL-norleucine, adipate and sucrose; nitrate reduction; growth in the presence of 0.5% NaCl; and beta galactosidase activity. PMID- 11411679 TI - 'Candidatus Phytoplasma brasiliense', a new phytoplasma taxon associated with hibiscus witches' broom disease. AB - Hibiscus rosa-sinensis is a valuable ornamental species widely planted in Brazil. Many plants are affected by witches' broom disease, which is characterized by excessive axillary branching, abnormally small leaves, and deformed flowers, symptoms that are characteristic of diseases attributed to phytoplasmas. A phytoplasma was detected in diseased Hibiscus by amplification of rRNA operon sequences by PCRs, and was characterized by RFLP and nucleotide sequence analyses of 16S rDNA. The collective RFLP patterns of amplified 16S rDNA differed from the patterns described previously for other phytoplasmas. On the basis of the RFLP patterns, the hibiscus witches' broom phytoplasma was classified in a new 16S rRNA RFLP group, designated group 16SrXV. A phylogenetic analysis of 16S rDNA sequences from this and other phytoplasmas identified the hibiscus witches' broom phytoplasma as a member of a distinct subclade (designated subclade xiv) of the class Mollicutes. A phylogenetic tree constructed on the basis of 16S rRNA gene sequences was consistent with the hypothesis that there was divergent evolution of hibiscus witches' broom phytoplasma and its closest relatives (members of 16S rRNA RFLP group 16SrII) from a common ancestor. On the basis of unique properties of the DNA from hibiscus witches' broom phytoplasma, it is proposed that it represents a new taxon, namely 'Candidatus Phytoplasma brasiliense'. PMID- 11411680 TI - Clostridium uliginosum sp. nov., a novel acid-tolerant, anaerobic bacterium with connecting filaments. AB - An anaerobic, acid-tolerant bacterium, CK55T, was isolated from an acidic forest bog. Cells of CK55T stained Gram-negative but did not have an outer membrane. Cells were spore-forming, motile rods with peritrichous flagella, formed chains or aggregates and were linked by connecting filaments that were composed of a core and outer sheath. Cellobiose, glucose, xylose, mannose, mannitol, sucrose and peptone supported growth. Arabinose, lactose, raffinose, H2/CO2, CO/CO2, vanillate, Casamino acids and various purines and pyrimidines did not support growth. Growth on carbohydrates yielded acetate, butyrate, lactate, formate and H2 as end-products. Growth was observed at pH 4.0-9.0, with an optimum at pH 6.5, and at 10-30 degrees C, with an optimum at 20-25 degrees C. At 20 degrees C, doubling times were 4 and 6 h at pH 6.5 and 4.0, respectively. Hydrogenase activity in cell-free extracts was 12 U (mg protein)-1. CK55T did not: (i) contain detectable levels of CO, formate, lactate dehydrogenases or cytochromes; (ii) carry out dissimilatory reduction of nitrate or sulfate; or (iii) produce methane. Thus, CK55T was characterized as a non-acetogenic, fermentative chemo organotroph. The G+C content of CK55T was 28.0 mol%. CK55T was phylogenetically most closely related to Clostridium botulinum (types B, E and F), Clostridium acetobutylicum and other saccharolytic clostridia; the 16S rRNA gene sequence similarity values to the nearest relatives of CK55T were approximately 97%. Based on morphological, physiological and phylogenetic properties of CK55T, it is proposed that CK55T be termed Clostridium uliginosum sp. nov. (= DSM 12992T = ATCC BAA-53T). PMID- 11411681 TI - Transfer of thermobacteroides leptospartum and Clostridium thermolacticum as Clostridium stercorarium subsp. leptospartum subsp. thermolacticum subsp. nov., comb. nov. and C. stercorarium subsp. thermolacticum subsp. nov., comb. nov. AB - 16S rRNA sequencing and sequence analysis of the sole member of the genus Thermobacteroides, Thermobacteroides leptospartum, revealed that it was related to members of cluster III (according to the scheme of Collins et al. 1994) represented exclusively by cellulolytic Clostridium species. Phenotypic studies indicated that Thermobacteroides leptospartum was also able to grow on cellulose, providing further evidence of its affiliation to members of cluster III. Its closest phylogenetic relatives, Clostridium thermolacticum and Clostridium stercorarium, were almost equidistantly placed with a similarity value of 99%. DNA hybridization studies also indicated that Thermobacteroides leptospartum, C. thermolacticum and C. stercorarium were closely related to each other (values of over 95% homology). Similarities based on the comparison of the 16S rRNA gene sequences and DNA homology are sufficiently high to regard all three strains as subspecies of a single species. It is therefore proposed that Thermobacteroides leptospartum and C. thermolacticum be transferred to cluster III as C. stercorarium subsp. leptospartum subsp. nov., comb. nov. and C. stercorarium subsp. thermolacticum subsp. nov., comb. nov., respectively, thus automatically creating C. stercorarium subsp. stercorarium subsp. nov., comb. nov. The transfer of the sole member of Thermobacteroides invalidates the taxonomic status of the genus. PMID- 11411682 TI - Transfer of Natrialba asiatica B1T to Natrialba taiwanensis sp. nov. and description of Natrialba aegyptiaca sp. nov., a novel extremely halophilic, aerobic, non-pigmented member of the Archaea from Egypt that produces extracellular poly(glutamic acid). AB - A novel extremely halophilic member of the Archaea, strain 40T, was isolated from Egypt (Aswan). This isolate requires at least 1.6 M sodium chloride for growth and exhibits optimal growth between 37 and 42 degrees C. Determination of the entire 16S rRNA gene sequence revealed the highest similarity to the type strain of Natrialba asiatica (> 99%). Polar lipid analysis indicated that strain 40T and Natrialba asiatica have essentially identical compositions, indicating that the former is a member of genus Natrialba. However, physiological and biochemical data provided evidence that Natrialba asiatica strains B1T and 172P1T, as well as strain 40T, are sufficiently different to be divided in three different species. The G+C content of strain 40T was 61.5+/-0.6 mol%. In addition, DNA-DNA hybridization data supported the placement of the isolate in a new species in the genus Natrialba, Natrialba aegyptiaca sp. nov., and indicated that Natrialba asiatica strain B1T should also be placed in a separate species, Natrialba taiwanensis sp. nov. Morphological studies of strain 40T indicated clearly that this isolate appears in three completely different cell shapes (cocci, rods, tetrads) under different conditions of growth, including different sodium chloride concentrations and different growth temperatures. Another interesting property of strain 40T is the ability to produce an extracellular polymer, which was found to be composed predominantly of glutamic acid (85% w/w), representing poly(glutamic acid), carbohydrates (12.5% w/w) and unidentified compounds (2.5% w/w). Among the Archaea, production of an extracellular polysaccharide has been described for some members of the genera Haloferax and Haloarcula. PMID- 11411683 TI - Streptococcus ovis sp. nov., isolated from sheep. AB - Seven strains of an unknown Gram-positive catalase-negative chain-forming coccus shaped organism isolated from clinical specimens from sheep were characterized by phenotypic and molecular taxonomic methods. Comparative 16S rRNA gene sequencing studies demonstrated that the bacterium represents a new sub-line within the genus Streptococcus. The unknown bacterium was readily distinguished from recognized streptococcal species by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Streptococcus ovis sp. nov. The type strain of Streptococcus ovis is CCUG 39485T (= LMG 19174T). PMID- 11411684 TI - Legionella drozanskii sp. nov., Legionella rowbothamii sp. nov. and Legionella fallonii sp. nov.: three unusual new Legionella species. AB - Seven strains of Legionella-like amoebal pathogens (LLAPs) were characterized on the basis of their cultural and staining characteristics, biochemical reactions, serology, cellular fatty acids (CFAs), isoprenoid quinone composition, total DNA relatedness, analysis of 16S rRNA and macrophage infectivity potentiator (mip) gene sequence analyses. All seven strains exhibited limited growth on buffered charcoal yeast extract alpha (BCYE) agar, required cysteine for growth and contained branched-chain CFAs and quinones typical of Legionella species. The bacilli were Gram-negative and catalase-positive. There were varying degrees of serological cross-reactions between these LLAP strains and other previously described Legionella species. Results from the various tests revealed that four LLAP strains represent three unusual new species of Legionella: Legionella drozanskii sp. nov., type strain LLAP-1T; Legionella rowbothamii sp. nov., type strain LLAP-6T; and Legionella fallonii sp. nov., type strain LLAP-10T. Three other LLAP strains, designated LLAP-7FL, LLAP-7NF and LLAP-9, were shown to be members of the species Legionella lytica. The deductions made from the phenetic characteristics of these bacteria were consistent with the phylogenetic relationships inferred from 16S rRNA and mip gene sequence analyses. This study is the first to speciate LLAP strains on the basis of data including quantitative DNA hybridization. PMID- 11411685 TI - Halomonas marisflavae sp. nov., a halophilic bacterium isolated from the Yellow Sea in Korea. AB - A halophilic Gram-negative bacterial strain, SW32T, which was isolated from a sample from the Yellow Sea of Korea, was subjected to a polyphasic taxonomic study. This organism grew optimally in the presence of 0.5-12% NaCl. On the basis of phenotypic and phylogenetic data, strain SW32T appeared to be a member of the genus Halomonas. Strain SW32T formed a distinct evolutionary lineage within the phylogenetic clade comprising Halomonas species and the genera Zymobacter and Carnimonas. The 16S rDNA sequence of strain SW32T contains 19 signature characteristics of the genus Halomonas and the family Halomonadaceae. Strain SW32T possessed a single polar flagellum, ubiquinone-9 as the predominant respiratory lipoquinone and C18:1, C16:0 and C16:1 omega7c and/or iso-C15:0 20H as the major fatty acids. The DNA G+C content was 59 mol%. Levels of 16S rDNA similarity between strain SW32T and the type strains of all validly described Halomonas species were 92.0-93.8%. Strain SW32T exhibited 16S rDNA similarity values of 92.7% to Zymobacter palmae IAM 14233T and 91.6% to Carnimonas nigrificans CECT 4437T. These data indicate that strain SW32T was related enough to members of the genus Halomonas to be placed as a new species within that genus. Therefore the name Halomonas marisflavae sp. nov. is proposed for strain SW32T. The type strain of the new species is strain SW32T (= KCCM 80003T = JCM 10873T). PMID- 11411686 TI - Porphyromonas gulae sp. nov., an anaerobic, gram-negative coccobacillus from the gingival sulcus of various animal hosts. AB - A new species, Porphyromonas gulae sp. nov., is proposed to include strains isolated from the gingival sulcus of various animal hosts which are distinct from related strains of Porphyromonas gingivalis of human origin. This bacterium exhibits the following characteristics: black-pigmented colonies; asaccharolytic, obligate anaerobic growth; and Gram-negative, non-motile and non-spore-forming, rod-shaped cells. Colonies do not fluoresce under UV light. Vitamin K1 and haemin are required for growth. Cells haemagglutinate sheep erythrocytes. Major fatty acid end products are butyric acid, isovaleric acid, succinic acid and phenylacetic acid. Strains are catalase-positive and indole is produced. Alkaline phosphatase, trypsin-like and N-acetyl-beta-glucosaminidase activities are strong. A beta-galactosidase and a glutamylglutamic acid arylamidase are also present. The G+C content of the chromosomal DNA is 51 mol%. DNA-DNA homology data and 16S rRNA gene sequence analysis provide strong evidence that strains from the animal biotype of P. gingivalis represent a Porphyromonas species that is distinct from P. gingivalis. The type strain of P. gulae is Loup 1T (= ATCC 51700T = NCTC 13180T). PMID- 11411687 TI - Molecular phylogenetics of the genus Rhodotorula and related basidiomycetous yeasts inferred from the mitochondrial cytochrome b gene. AB - Phylogenetic relationships of basidiomycetous yeasts, especially of the genus Rhodotorula, were studied using partial sequences of the mitochondrial cytochrome b gene. The results demonstrated that the basidiomycetous yeasts under investigation distributed into two main clusters: one containing Tremellales, Filobasidiales and their anamorphs and the other containing Ustilaginales, Sporidiales and their anamorphs. This clustering in turn correlates with cell wall biochemistry, presence or absence of xylose, and septal ultrastructure, dolipore or simple pore. Bullera, Bulleromyces, Filobasidiella, Cryptococcus and Trichosporon, yeasts of the former cluster, contain xylose in the cell wall and have dolipore septa. In contrast yeasts of the latter cluster, which included Bensingtonia, Erythrobasidium, Leucosporidium, Malassezia, Rhodosporidium, Rhodotorula, Sporidiobolus, Sporobolomyces and Ustilago, have no xylose in the cell wall and have a simple pore septum. Yeasts of the latter group could be further divided into four clades (A-D). Species of Rhodotorula were distributed in all of these clades, indicating the polyphyletic nature of the genus. A limited number of Rhodotorula species demonstrated identical sequences, for example Rhodotorula bacarum and Rhodotorula foliorum, Rhodotorula fujisanensis and Rhodotorula futronensis, Rhodotorula glutinis var. dairenensis and Rhodotorula mucilaginosa. However, all the other test species of the genus Rhodotorula were well separated based on their 396 bp nucleotide sequences. These results demonstrate the effectiveness of the use of cytochrome b sequences for both species identification and the study of phylogenetic relationships among basidiomycetous yeasts. PMID- 11411688 TI - Candida cleridarum, Candida tilneyi and Candida powellii, three new yeast species isolated from insects associated with flowers. AB - Three new asexual yeast species were isolated from various floricolous insects. Candida cleridarum sp. nov. was the dominant species in clerid beetles collected in flowers of various cacti in Arizona and Southern California. The sequence of the D1D2 domains of the large-subunit rDNA showed that it is a sister species to Candida fragi (0.9% base difference), a yeast isolated once from fermenting strawberries. Candida tilneyi sp. nov. and Candida powellii sp. nov. were recovered from bees and from nitidulid beetles in flowers of two species of morning glory (Ipomoea) in north-western Costa Rica. C. tilneyi sp. nov. is most closely related to Candida geochares, but differs in the D1D2 sequence by 4.7% base substitutions. C. powellii sp. nov. is a relative of Candida batistae and Candida floricola, showing sequence differences of 5.9 and 6.9%, respectively. In all cases, the new species are phenotypically similar to their nearest relatives, but are sufficiently different to allow conventional identification. The type strains are C. cleridarum strain UWO(PS) 99-101.1T ( = CBS 8793T), C. tilneyi strain UWO(PS) 99-325.1T ( = CBS 8794T) and C. powellii strain UWO(PS) 99-325.3T ( = CBS 8795T). PMID- 11411689 TI - Bensingtonia thailandica sp. nov., a novel basidiomycetous yeast species isolated from plant leaves in Thailand. AB - Ten strains which were characterized by the formation of ballistoconidia, the absence of xylose in whole-cell hydrolysates, the presence of Q-9 as the major ubiquinone isoprenologue, the inability to ferment sugars and positive diazonium blue B and urease reactions were isolated from plant samples collected in Thailand. These isolates were closely related to Bensingtonia phyllada based on the analysis of 18S rDNA sequences. On the basis of the morphological, physiological and chemotaxonomic properties, the 10 isolates were assigned to the genus Bensingtonia. DNA complementarity showed that these isolates were genetically distinct from known species of the genus Bensingtonia. The isolates are described as Bensingtonia thailandica sp. nov. The type strain is strain TY 138T (= JCM 10651T). PMID- 11411690 TI - Candida sorbosivorans sp. nov., a new member of the genus Candida Berkhout. AB - A yeast, strain NCYC 2938T, was isolated from contaminated industrial material. This material was involved in a cascade continuous process for oxidizing sorbitol (D-glucitol) to L-sorbose. The isolate is similar, although not identical, to Candida geochares and Candida magnoliae in its physiological characteristics. Sequence analysis of the 26S rDNA D1/D2 variable domain showed that it was similar to those of both Candida species, but differed sufficiently to be considered as a separate species. Both the physiological characteristics and the unique 26S rDNA D1/D2 sequence of NCYC 2938T are described here, and the yeast has been named Candida sorbosivorans sp. nov. The type strain is NCYC 2938T (= CBS 8768T). PMID- 11411691 TI - Two new yeasts, Trichosporon debeurmannianum sp. nov. and Trichosporon dermatis sp. nov., transferred from the Cryptococcus humicola complex. AB - Cryptococcus humicola, as currently defined, shows intraspecific rRNA gene sequence differences. Three strains of this species produced arthroconidia on cornmeal agar and belonged to the genus Trichosporon in a molecular phylogeny. They clustered with the species possessing Q10 as the major ubiquinone and were serotype I. Sequence analyses clearly revealed that they were two new Trichosporon species. The names Trichosporon dermatis sp. nov. (= CBS 2043T) and Trichosporon debeurmannianum sp. nov. (= CBS 1896T) are proposed for these strains. PMID- 11411692 TI - Construction and bootstrap analysis of DNA fingerprinting-based phylogenetic trees with the freeware program FreeTree: application to trichomonad parasites. AB - The Win95/98/NT program FreeTree for computation of distance matrices and construction of phylogenetic or phenetic trees on the basis of random amplified polymorphic DNA (RAPD), RFLP and allozyme data is presented. In contrast to other similar software, the program FreeTree (available at http://www.natur.cuni.cz/~flegr/programs/freetree or http://ijs.sgmjournals.org/content/vol51/issue3/) can also assess the robustness of the tree topology by bootstrap, jackknife or operational taxonomic unit jackknife analysis. Moreover, the program can be also used for the analysis of data obtained in several independent experiments performed with non-identical subsets of taxa. The function of the program was demonstrated by an analysis of RAPD data from 42 strains of 10 species of trichomonads. On the phylogenetic tree constructed using FreeTree, the high bootstrap values and short terminal branches for the Tritrichomonas foetus/suis 14-strain branch suggested relatively recent and probably clonal radiation of this species. At the same time, the relatively lower bootstrap values and long terminal branches for the Trichomonas vaginalis 20-strain branch suggested more ancient radiation of this species and the possible existence of genetic recombination (sexual reproduction) in this human pathogen. The low bootstrap values and the star-like topology of the whole Trichomonadidae tree confirm that the RAPD method is not suitable for phylogenetic analysis of protozoa at the level of higher taxa. It is proposed that the repeated bootstrap analysis should be an obligatory part of any RAPD study. It makes it possible to assess the reliability of the tree obtained and to adjust the amount of collected data (the number of random primers) to the amount of phylogenetic signals in the RAPD data of the taxon analysed. The FreeTree program makes such analysis possible. PMID- 11411693 TI - Phylogenetic relationships among algae based on complete large-subunit rRNA sequences. AB - The complete or nearly complete large-subunit rRNA (LSU rRNA) sequences were determined for representatives of several algal groups such as the chlorarachniophytes, cryptomonads, haptophytes, bacillariophytes, dictyochophytes and pelagophytes. Our aim was to study the phylogenetic position and relationships of the different groups of algae, and in particular to study the relationships among the different classes of heterokont algae. In LSU rRNA phylogenies, the chlorarachniophytes, cryptomonads and haptophytes seem to form independent evolutionary lineages, for which a specific relationship with any of the other eukaryotic taxa cannot be demonstrated. This is in accordance with phylogenies inferred on the basis of the small-subunit rRNA (SSU rRNA). Regarding the heterokont algae, which form a well-supported monophyletic lineage on the basis of LSU rRNA, resolution between the different classes could be improved by combining the SSU and LSU rRNA data. Based on a concatenated alignment of both molecules, the phaeophytes and the xanthophytes are sister taxa, as well as the pelagophytes and the dictyochophytes, and the chrysophytes and the eustigmatophytes. All these sister group relationships are highly supported by bootstrap analysis and by different methods of tree construction. PMID- 11411694 TI - Phylogenetic position and inter-relationships of the osmotrophic euglenids based on SSU rDNA data, with emphasis on the Rhabdomonadales (Euglenozoa). AB - In order to reconstruct the evolution of euglenid flagellates, euglenozoan SSU rDNA data have been used to investigate phylogenetic relationships with a focus on osmotrophic taxa and especially on the Rhabdomonadales. The dataset consisting of the SSU rDNAs of osmotrophic, phagotrophic and phototrophic taxa was used in parsimony, maximum-likelihood and distance analyses. Five genera make up the Rhabdomonadales, all of them osmotrophic: Gyropaigne, Menoidium, Parmidium, Rhabdomonas and Rhabdospira. According to our analyses they form a strongly supported monophyletic assemblage which is characterized by a low sequence divergence compared to the euglenids in general. Closest relatives are the members of the osmotrophic genus Distigma. All primary osmotrophic species constitute a larger monophyletic group with the phototrophic euglenids and the phagotroph Peranema trichophorum. The combination of three rhabdomonadalian species Rhabdomonas gibba, Rhabdomonas spiralis and Rhabdospira spiralis with nearly identical SSU rDNA sequences is strongly recommended. The phagotroph Petalomonas cantuscygni branches at the bottom of the euglenid subtree with significantly weaker support. The inter-relationship of the three distinct euglenozoan taxa (euglenids, kinetoplastids and diplonemids) could not be convincingly resolved by this study. PMID- 11411695 TI - Phragmoplast-mediated cytokinesis in Trentepohlia: results of TEM and immunofluorescence cytochemistry. AB - This paper reports transmission electron microscopy (TEM) and immunofluorescence evidence for phragmoplast-mediated cytokinesis in the green alga, Trentepohlia (Ulvophyceae, Chlorophyta). This type of cytokinesis is normally found only in land plants and some charophycean green algae. Earlier TEM work documented a phragmoplast in another trentepohlialean alga, Cephaleuros. Numerous molecular studies have shown that both Trentepohlia and Cephaleuros are clearly not in the charophycean (streptophyte) lineage of green algae. Also, details of vegetative cytokinesis in Trentepohlia and Cephaleuros indicate differences from the processes and structures observed in the streptophytes. Parallel evolution could be the explanation for a phragmoplast-mediated cytokinesis in both the chlorophycean Trentepohliales and the streptophyte lineage of charophycean green algae and land plants. PMID- 11411696 TI - Phylogenetic position of a renal coccidium of the European green frogs, 'Isospora' lieberkuehni Labbe, 1894 (Apicomplexa: Sarcocystidae) and its taxonomic implications. AB - 'Isospora' lieberkuehni, an unusual isosporoid renal coccidium that parasitizes the European water frog was isolated from the edible frog, Rana kl. esculenta, in the Czech Republic. Sequencing of the small-subunit (SSU) rRNA gene showed that it belongs to the family Sarcocystidae, being closely related to a clade comprising members of the subfamily Toxoplasmatinae. The position within Sarcocystidae correlates with the mode of excystation via collapsible plates as postulated by previous authors. Phylogenetic, morphological and biological differences between 'Isospora' lieberkuehni and the other Stiedabody-lacking members of the genus Isospora justify separation of this coccidium on a generic level. Hyaloklossia Labbe, 1896 is the oldest available synonym and is herein re erected. The original definition of the genus Hyaloklossia is emended based on recent observations. PMID- 11411697 TI - Phylogenetic analysis of chloroplast small-subunit rRNA genes of the genus Euglena Ehrenberg. AB - Almost complete sequences of plastid SSU rDNA (16S rDNA) from 17 species belonging to the order Euglenales (sensu Nemeth, 1997; Shi et al., 1999) were determined and used to infer phylogenetic relationships between 10 species of Euglena, three of Phacus, and one of each of Colacium, Lepocinclis, Strombomonas, Trachelomonas and Eutreptia. The maximum-parsimony (MP), maximum-likelihood (ML) and distance analyses of the unambiguously aligned sequence fragments imply that the genus Euglena is not monophyletic. Parsimony and distance methods divide Euglenaceae into two sister groups. One comprises of representatives from the subgenera Phacus, Lepocinclis and Discoglena (sensu Zakrys, 1986), whereas the other includes members of Euglena and Calliglena subgenera (sensu Zakrys, 1986), intermixed with representatives of Colacium, Strombomonas and Trachelomonas. In all analyses subgenera Euglena--together with Euglena polymorpha (representative of the subgenus Calliglena)--and Discoglena--together with Phacus and Lepocinclis -form two well-defined clades. The data clearly indicate that a substantial revision of euglenoid systematics is very much required, nevertheless it must await while more information can be gathered, allowing resolution of outstanding relationships. PMID- 11411699 TI - Validation of publication of new names and new combinations previously effectively published outside the IJSEM. PMID- 11411698 TI - Phylogenetic analysis of phagotrophic, photomorphic and osmotrophic euglenoids by using the nuclear 18S rDNA sequence. AB - Phylogenetic analyses of 35 strains including 25 previously published sequences and 10 which have been newly sequenced, representing two species of Euglena, five species of Phacus and three species of Astasia, were carried out using the SSU rDNA. Parsimony, distance and maximum-likelihood inferred phylogenies support (1) monophyly of the euglenoids, (2) kinetoplastids as the sister group, (3) the phagotrophic Petalomonas cantuscygni Cann et Pennick anchoring the base of the euglenoid lineage, (4) evolution of phototrophy within the euglenoids from a single event, (5) multiple origins of osmotrophic euglenoids and (6) polyphyly of the genera Euglena Ehrenberg and Phacus Dujardin. Analyses also indicate that Lepocinclis Perty, Trachelomonas Ehrenberg and Astasia Dujardin are polyphyletic. In addition, the results suggest that neither the Euglenales nor the Eutreptiales form a monophyletic lineage, thus questioning currently available classifications. Concerning the phagotrophic mode of nutrition, the data suggest that the feeding apparatus arose multiple times. PMID- 11411700 TI - Notification that new names and new combinations have appeared in volume 50, part 2, of the IJSEM. PMID- 11411701 TI - The taxonomy of Streptomyces and related genera. AB - The streptomycetes, producers of more than half of the 10,000 documented bioactive compounds, have offered over 50 years of interest to industry and academia. Despite this, their taxonomy remains somewhat confused and the definition of species is unresolved due to the variety of morphological, cultural, physiological and biochemical characteristics that are observed at both the inter- and the intraspecies level. This review addresses the current status of streptomycete taxonomy, highlighting the value of a polyphasic approach that utilizes genotypic and phenotypic traits for the delimitation of species within the genus. PMID- 11411702 TI - 'Candidatus Mycoplasma haemominutum', a low-virulence epierythrocytic parasite of cats. AB - The phylogenetic position and some taxonomically relevant characteristics of a small, low-virulence bacterial parasite of cats are described. A 16S rDNA analysis revealed that the organism was in the Mycoplasma clade and was most closely related to a parasite of pigs previously designated Eperythrozoon suis. As the organism has not been cultured in vitro and is maintained in serial passage in cats in vivo, Candidatus status is proposed for this novel taxon as 'Candidatus Mycoplasma haemominutum'. PMID- 11411703 TI - Differentiation between Campylobacter hyoilei and Campylobater coli using genotypic and phenotypic analyses. AB - Genotypic and phenotypic methods were applied to investigate differences between the closely related species Campylobacter hyoilei and Campylobacter coli. A unique DNA sequence from C. hyoilei was used to design a specific PCR assay that amplified a DNA product of 383 bp for all C. hyoilei strains, but not other Campylobacter species, including C. coli. The PCR assay could detect 100 fg pure C. hyoilei DNA, 2 x 10(2) c.f.u. ml(-1) using cultured cells and 8.3 x 10(3) c.f.u. 0.1 g(-1) in faeces. The C. hyoilei sequence utilized for specific detection and identification of this species showed similarities to sequences from bacteriophages Mu, P2 and 186, suggesting lysogination of the ancestral C. hyoilei genome. Activities of a set of 15 enzymes that participate in a variety of cellular functions, including biosynthesis, catabolism, energy generation, maintenance of redox balance and phosphate utilization, were tested using sets of strains of C. hyoilei and C. coli. Comparison of mean rates of enzyme activities revealed significant differences between species in the values determined for seven of these activities. Both the genetic and phenotypic data indicate that C. hyoilei is a unique Campylobacter species. PMID- 11411704 TI - Sphingomonas pituitosa sp. nov., an exopolysaccharide-producing bacterium that secretes an unusual type of sphingan. AB - Strain EDIVT, an exopolysaccharide-producing bacterium, was subjected to polyphasic characterization. The bacterium produced copious amounts of an extracellular polysaccharide, forming slimy, viscous, intensely yellow-pigmented colonies on Czapek-Dox (CZD) agar. The culture fluids of the liquid version of CZD medium were highly viscous after cultivation for 5 d. Cells of strain EDIVT were Gram-negative, catalase-positive, oxidase-negative, nonspore-forming, rod shaped and motile. Comparisons of 16S rDNA gene sequences demonstrated that EDIVT clusters phylogenetically with the species of the genus Sphingomonas sensu stricto. The G+C content of the DNA (64.5 mol%), the presence of ubiquinone Q-10, the presence of 2-hydroxymyristic acid (14:0 2-OH) as the major hydroxylated fatty acid, the absence of 3-hydroxy fatty acids and the detection of sym homospermidine as the major component in the polyamine pattern, together with the presence of sphingoglycolipid, supported this delineation. 16S rDNA sequence analysis indicated that strain EDIVT is most closely related (99.4% similarity) to Sphingomonas trueperi LMG 2142T. DNA-DNA hybridization showed that the level of relatedness to S. trueperi is only 45.5%. Further differences were apparent in the cellular fatty acid profile, the polar lipid pattern, the Fourier-transform infrared spectrum and whole-cell proteins and in a number of biochemical characteristics. On the basis of the estimated phylogenetic position derived from 16S rDNA sequence data, DNA-DNA reassociation and phenotypic differences, strain EDIVT (= CIP 106154T = DSM 13101T) was recognized as a new species of Sphingomonas, for which the name Sphingomonas pituitosa sp. nov. is proposed. A component analysis of the exopolysaccharide (named PS-EDIV) suggested that it represents a novel type of sphingan composed of glucose, rhamnose and an unidentified sugar. Glucuronic acid, which is commonly found in sphingans, was absent. The mean molecular mass of PS-EDIV was approximately 3 x 10(6) Da. PMID- 11411705 TI - Corynebacterium mooreparkense sp. nov. and Corynebacterium casei sp. nov., isolated from the surface of a smear-ripened cheese. AB - Ten isolates each of two different bacterial species isolated from the surface of a smear-ripened cheese were found to exhibit many characteristics of the genus Corynebacterium. The isolates were Gram-positive, catalase-positive, non-spore forming rods that did not undergo a rod/coccus transformation when grown on complex media. Chemotaxonomic investigation revealed that the strains belonged unambiguously to the genus Corynebacterium. Their cell walls contained arabinose, galactose and short-chain mycolic acids (C22 to C36) and their peptidoglycan contained meso-diaminopimelic acid. The G+C content of the DNA was 51-60 mol%. MK 9 (H2) was the principal menaquinone. The 16S rDNA sequences of four isolates of each bacterium were determined and aligned with those of other members of the coryneform group. Phylogenetic analysis showed that the strains represented two new sublines within the genus Corynebacterium; Corynebacterium variabile and Corynebacterium ammoniagenes were their nearest known phylogenetic neighbours. Corynebacterium variabile and Corynebacterium ammoniagenes showed the highest levels of sequence homology with the isolates; however, DNA-DNA hydridization studies indicated that the Corynebacterium strains isolated from the cheese smear did not belong to either Corynebacterium variabile or Corynebacterium ammoniagenes (26 and 46% chromosomal similarity, respectively). On the basis of the phylogenetic and phenotypic distinctiveness of the unknown isolates, it is proposed that the bacteria be classified as two new Corynebacterium species, for which the names Corynebacterium mooreparkense sp. nov. and Corynebacterium casei sp. nov. are proposed. Type strains have been deposited in culture collections as Corynebacterium mooreparkense LMG S-19265T (= NCIMB 30131T) and Corynebacterium casei LMG S-19264T (= NCIMB 30130T). PMID- 11411706 TI - Actinomyces funkei sp. nov., isolated from human clinical specimens. AB - Three strains of a previously undescribed Actinomyces-like bacterium were isolated from human clinical specimens. Phenotypic studies indicated that the strains were members of the genus Actinomyces and were presumptively identified as Actinomyces turicensis. Comparative 16S rRNA gene sequencing studies showed that although the bacterium is phylogenetically closely related to Actinomyces turicensis, it nevertheless constitutes a new sub-line within the genus Actinomyces. Based on phenotypic and molecular chemical and molecular genetic evidence, it is proposed that the unknown Actinomyces-like bacterium from human clinical specimens be classified as Actinomyces funkei sp. nov. The type strain of Actinomyces funkei is CCUG 42773T (= CIP 106713T). PMID- 11411707 TI - Corynebacterium capitovis sp. nov., from a sheep. AB - An unknown Gram-positive rod-shaped bacterium was isolated from skin scrapings from the infected head of a sheep and subjected to a polyphasic taxonomic analysis. Chemical analysis revealed the presence of straight-chain and monounsaturated fatty acids and short-chain (C32-C36) mycolic acids consistent with the genus Corynebacterium. Comparative 16S rRNA gene sequencing confirmed that the unknown rod was a member of the genus Corynebacterium, with the organism forming a distinct sub-line and displaying greater than 3% sequence divergence with established species. The unknown Corynebacterium isolate was readily distinguished from recognized species of the genus by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium from a sheep be classified in the genus Corynebacterium, as Corynebacterium capitovis sp. nov. The type strain of Corynebacterium capitovis is CCUG 39779T (= CIP 106739T). PMID- 11411708 TI - Phylogenetic analyses of Synechococcus strains (cyanobacteria) using sequences of 16S rDNA and part of the phycocyanin operon reveal multiple evolutionary lines and reflect phycobilin content. AB - The genus Synechococcus (cyanobacteria), while containing morphologically similar isolates, is polyphyletic and organisms presently classified as such require reclassification into several independent genera. Studies based on analysis of 16S rRNA gene sequences have shown that members of the genus Synechococcus are affiliated to three of seven deeply branching cyanobacterial lineages. In addition, some strains do not appear to be associated with any of these lineages and may represent novel clades. In this report, a cyanobacterial phylogeny based on 16S rDNA sequences, including 14 newly sequenced Synechococcus isolates, is presented. One newly sequenced Synechococcus strain (PCC 7902) did not have any close relatives amongst cyanobacterial isolates currently contained in 16S rDNA sequence databases and was only loosely affiliated to a cyanobacterial lineage in which no other Synechococcus strains were found. Three hot-spring Synechococcus isolates, including two that were newly sequenced in this study (PCC 6716 and PCC 6717), formed an additional cyanobacterial lineage. These results indicated that Synechococcus species are affiliated to five of eight deeply branching cyanobacterial lineages. Part of the phycocyanin (PC) gene sequence (cpc), including the intergenic spacer (IGS) between cpcB and cpcA and the corresponding flanking regions (cpcBA-IGS), was used to investigate relationships between closely related Synechococcus isolates. Previously described PCR primers did not amplify this region from the majority of strains under investigation, so a new set of primers was designed that allowed amplification and sequencing of the cpcBA-IGS and flanking regions from 38 Synechococcus species. Phylogenetic analysis of this region was largely consistent with that obtained from 16S rDNA sequence analysis and revealed a relationship between the primary PC DNA sequence and the phycobilin content of cells. PMID- 11411709 TI - A proposal for the unification of five species of the cyanobacterial genus Microcystis Kutzing ex Lemmermann 1907 under the rules of the Bacteriological Code. AB - Genomic DNA homologies were examined from six Microcystis (cyanobacteria) strains, including five different species, Microcystis aeruginosa, Microcystis ichthyoblabe, Microcystis novacekii, Microcystis viridis and Microcystis wesenbergii. All DNA-DNA reassociation values between two strains of M. aeruginosa and the other four species exceeded 70%, which is considered high enough for them to be classified within the same bacterial species. It is proposed to unify these five species into M. aeruginosa under the Rules of the Bacteriological Code and NIES843T (= IAM M-247T) is proposed as the type strain. Two other species, Microcystis flos-aquae and Microcystis pseudofilamentosa, should be regarded as morphological variations of this unified M. aeruginosa. The current taxonomy of cyanobacteria depends too much upon morphological characteristics and must be reviewed by means of bacteriological methods as well as traditional botanical methods. PMID- 11411710 TI - Proposal to transfer Actinomadura carminata to a new subspecies of the genus Nonomuraea as Nonomuraea roseoviolacea subsp. carminata comb. nov. AB - An anthracycline-producing actinomycete (strain SF2303) was compared with two other anthracycline producers, Actinomadura carminata IFO 15903T and Nonomuraea roseoviolacea IFO 14098T, using morphological, physiological, chemotaxonomic and molecular-genetic criteria. The morphological and cultural characteristics of these three strains are similar. The physiological properties of strain SF2303 and N. roseoviolacea IFO 14098T are very similar, but are different from those of A. carminata IFO 15903T in the utilization of some sugars and the reduction of nitrate. Their chemotaxonomic properties [cell wall chemotype, IIIB; major menaquinone, MK-9 (III, VIIl-H4); phospholipid type, PIV; cellular fatty acids 10M-17:0/17:1 and iso-16:0 as major components and 2-hydroxy fatty acids as minor components; mycolic acid, absent] were identical and indicated that these three strains belong to the family Streptosporangiaceae. On the basis of 16S rDNA sequences and phylogenetic analysis, they were all included in the cluster formed by species of Nonomuraea. The levels of DNA relatedness between strain SF2303 and N. roseoviolacea IFO 14098T ranged from 71 to 78%; however, the levels of relatedness between the two strains and A. carminata IFO 15903T were lower (49 60%). Therefore, strain SF2303 was identified as a member of the species N. roseoviolacea and it is proposed that Actinomadura carminata Gauze et al. 1973 should be transferred to a new subspecies of the genus Nonomuraea Zhang et al. 1998 as N. roseoviolacea subsp. carminata comb. nov. PMID- 11411711 TI - Proposal to transfer some members of the genera Haemobartonella and Eperythrozoon to the genus Mycoplasma with descriptions of 'Candidatus Mycoplasma haemofelis', 'Candidatus Mycoplasma haemomuris', 'Candidatus Mycoplasma haemosuis' and 'Candidatus Mycoplasma wenyonii'. AB - Cell-wall-less uncultivated parasitic bacteria that attach to the surface of host erythrocytes currently are classified in the order Rickettsiales, family Anaplasmataceae, in the genera Haemobartonella and Eperythrozoon. Recently 16S rRNA gene sequences have been determined for four of these species: Haemobartonella felis and Haemobartonella muris and Eperythrozoon suis and Eperythrozoon wenyonii. Phylogenetic analysis of these sequence data shows that these haemotrophic bacteria are closely related to species in the genus Mycoplasma (class Mollicutes). These haemotrophic bacteria form a new phylogenetic cluster within the so-called pneumoniae group of Mycoplasma and share properties with one another as well as with other members of the pneumoniae group. These studies clearly indicate that the classification of these taxa should be changed to reflect their phylogenetic affiliation and the following is proposed: (i) that Haemobartonella felis and Haemobartonella muris should be transferred to the genus Mycoplasma as 'Candidatus Mycoplasma haemofelis' and 'Candidatus Mycoplasma haemomuris' and (ii) that Eperythrozoon suis and Eperythrozoon wenyonii should be transferred to the genus Mycoplasma as 'Candidatus Mycoplasma haemosuis' and 'Candidatus Mycoplasma wenyonii'. The former Haemobartonella and Eperythrozoon species described here represent a new group of parasitic mycoplasmas that possess a pathogenic capacity previously unrecognized among the mollicutes. These haemotrophic mycoplasmas have been given the trivial name haemoplasmas. These results call into question the affiliation of the remaining officially named species of Haemobartonella and Eperythrozoon which should be considered species of uncertain affiliation pending the resolution of their phylogenetic status. PMID- 11411712 TI - Phylogenetic positions of Clostridium novyi and Clostridium haemolyticum based on 16S rDNA sequences. AB - The partial sequences (1465 bp) of the 16S rDNA of Clostridium novyi types A, B and C and Clostridium haemolyticum were determined. C. novyi types A, B and C and C. haemolyticum clustered with Clostridium botulinum types C and D. Moreover, the 16S rDNA sequences of C. novyi type B strains and C. haemolyticum strains were completely identical; they differed by 1 bp (level of similarity > 99.9%) from that of C. novyi type C, they were 98.7% homologous to that of C. novyi type A (relative positions 28-1520 of the Escherichia coli 16S rDNA sequence) and they exhibited a higher similarity to the 16S rDNA sequence of C. botulinum types D and C than to that of C. novyi type A. These results suggest that C. novyi types B and C and C. haemolyticum may be one independent species generated from the same phylogenetic origin. PMID- 11411713 TI - Transfer of [Pseudomonas] lemoignei, a gram-negative rod with restricted catabolic capacity, to Paucimonas gen. nov. with one species, Paucimonas lemoignei comb. nov. AB - The phylogenetic positions of the type strain and strain A62 of [Pseudomonas] lemoignei, as well as those of nine bacterial isolates closely related to [Pseudomonas] lemoignei, were re-evaluated by analysing morphological, physiological and molecular-biological characteristics. 16S rDNA analysis confirmed that the type strain of [Pseudomonas] lemoignei and strain A62 belong to the order 'Burkholderiales' of the beta-Proteobacteria and represent a new cluster, for which the name Paucimonas gen. nov. is proposed. Paucimonas contains a single species, Paucimonas lemoignei comb. nov. PMID- 11411714 TI - Rhizobium yanglingense sp. nov., isolated from arid and semi-arid regions in China. AB - A novel rhizobial group, cluster 9, defined in previous research [Tan, Z. Y., Wang, E. T., Peng, G. X., Zhu, M. E., Martinez-Romero, E. & Chen, W. X. (1999). Int J Syst Bacteriol 49, 1457-1469], was further characterized by determination of DNA base composition, whole-cell protein SDS-PAGE analysis, DNA-DNA hybridization, 16S rRNA gene sequencing and host specificity. These isolates were collected from the wild legumes Amphicarpaea trisperma, Coronilla varia and Gueldenstaedtia multiflora growing in arid and semi-arid regions in northwestern China. Isolates within cluster 9 grouped into a single cluster above a similarity level of 90.6% in a cluster analysis based on protein SDS-PAGE, and they were differentiated from defined rhizobial species. Comparative analysis of 16S rRNA gene sequences showed that isolate CCBAU 71623T, representing cluster 9, was most related to Rhizobium gallicum and Rhizobium mongolense. The DNA-DNA homologies were lower than 42.4% among cluster 9 and defined species, including R. gallicum and R. mongolense. These data indicated that cluster 9 was a unique genomic species. Isolates within this cluster could share their host plants. They could not nodulate Galega orientalis and Leucaena leucocephala and formed ineffective nodules on Phaseolus vulgaris. This group could also be differentiated from defined species by phenotypic characteristics. It is therefore proposed as a new species, Rhizobium yanglingense, with isolate CCBAU 71623 as the type strain. PMID- 11411715 TI - Phylogenetic diversity of Klebsiella pneumoniae and Klebsiella oxytoca clinical isolates revealed by randomly amplified polymorphic DNA, gyrA and parC genes sequencing and automated ribotyping. AB - The infra-specific phylogenetic diversity and genetic structure of both Klebsiella pneumoniae and Klebsiella oxytoca was investigated using a combination of randomly amplified polymorphic DNA (RAPD) analysis, sequencing of gyrA and parC genes, and automated ribotyping. After RAPD analysis with four independent primers of 120 clinical isolates collected from 22 European hospitals in 13 countries, K. pneumoniae isolates fell into three clusters and K. oxytoca isolates fell into two clusters, while Klebsiella planticola isolates formed a sixth cluster. Each cluster was geographically widespread. K. pneumoniae cluster I (KpI) accounted for 80% of the isolates of this species and included reference strains of the three subspecies K. pneumoniae subsp. pneumoniae, K. pneumoniae subsp. ozaenae and K. pneumoniae subsp. rhinoscleromatis. Clusters KpII and KpIII were equally represented, as were the two K. oxytoca clusters. Individualization of each cluster was fully confirmed by phylogenetic analysis of gyrA and parC gene sequences. In addition, sequence data supported the evolutionary separation of K. pneumoniae from a phylogenetic group including K. oxytoca, Klebsiella terrigena, K. planticola and Klebsiella ornithinolytica. Automated ribotyping using Mlu I appeared suitable for identification of each Klebsiella cluster. The adonitol fermentation test was found to be useful for cluster identification in K. pneumoniae, since it was negative in all strains of clusters KpIII and in some KpII strains, but always positive in cluster KpI. The usefulness of gyrA and parC sequence data for population genetics and cluster identification in bacteria was demonstrated, even for the phylogenetic positioning of quinolone-resistant isolates. PMID- 11411716 TI - Phylogenetic analyses of Klebsiella species delineate Klebsiella and Raoultella gen. nov., with description of Raoultella ornithinolytica comb. nov., Raoultella terrigena comb. nov. and Raoultella planticola comb. nov. AB - The phylogenetic relationships of the type strains of 9 Klebsiella species and 20 species from 11 genera of the family Enterobacteriaceae were investigated by performing a comparative analysis of the sequences of the 16S rRNA and rpoB genes. The sequence data were phylogenetically analysed by the neighbourjoining and parsimony methods. The phylogenetic inference of the sequence comparison confirmed that the genus Klebsiella is heterogeneous and composed of species which form three clusters that also included members of other genera, including Enterobacter aerogenes, Erwinia clusters I and II and Tatumella. Cluster I contained the type strains of Klebsiella pneumoniae subsp. pneumoniae, Klebsiella pneumoniae subsp. rhinoscleromatis and Klebsiella pneumoniae subsp. ozaenae. Cluster II contained Klebsiella ornithinolytica, Klebsiella planticola, Klebsiella trevisanii and Klebsiella terrigena, organisms characterized by growth at 10 degrees C and utilization of L-sorbose as carbon source. Cluster III contained Klebsiella oxytoca. The data from the sequence analyses along with previously reported biochemical and DNA-DNA hybridization data support the division of the genus Klebsiella into two genera and one genogroup. The name Raoultella is proposed as a genus name for species of cluster II and emended definitions of Klebsiella species are proposed. PMID- 11411717 TI - Nocardia veterana sp. nov., isolated from human bronchial lavage. AB - A nocardioform bacterium was isolated from the bronchoscopic lavage of a 78-year old man with a past history of tuberculous pleurisy, who presented with bilateral upper lobe lesions at Austin and Repatriation Medical Centre, Heidelberg, Australia. The strain was aerobic, Gram-positive, produced beige substrate mycelium and scant white aerial mycelium. It showed chemotaxonomic markers which were consistent with the classification of Nocardia: i.e. meso-diaminopimelic acid, N-glycolylmuramic acid, arabinose and galactose as diagnostic sugars; phospholipids phosphatidylinositol mannosides, phosphatidylinositol, phosphatidylethanolamine and diphosphatidylglycerol; a menaquinone with a cyclic isoprene side chain, MK-8(H4cycl.); a fatty acid pattern composed of unbranched saturated and monounsaturated fatty acids with a considerable amount of tuberculostearic acid; and mycolic acids composed of 54-62 carbon atoms with three principal mycolic acids which were mono- and polyunsaturated, showing a chain length C56, C58 and C60 and accounting for over 70% of the entire pattern. The 16S rDNA sequence showed the highest similarity to the type strain of Nocardia vaccinii; the DNA-DNA similarity of the two strains was 31%. These data, together with distinct physiological traits and molecular biological analyses, as well as chemotaxonomic results, led to the conclusion that the novel isolate represents a new species within the genus Nocardia for which the name Nocardia veterana sp. nov. is proposed. The type strain is M157222T (DSM 44445T = NRRL B 24136T). PMID- 11411718 TI - Proposal of Mycetocola gen. nov. in the family Microbacteriaceae and three new species, Mycetocola saprophilus sp. nov., Mycetocola tolaasinivorans sp. nov. and Mycetocola lacteus sp. nov., isolated from cultivated mushroom, Pleurotus ostreatus. AB - The taxonomic positions of 10 tolaasin-detoxifying bacteria, which were isolated from the cultivated mushroom Pleurotus ostreatus, were investigated. These strains are Gram-positive, obligately aerobic, non-sporulating and irregular rod shaped bacteria. They have the following characteristics: the major menaquinone is MK-10, the DNA G+C content ranges from 64 to 65 mol%, the diamino acid in the cell wall is lysine and the muramic acid in the peptidoglycan is an acetyl type. The major fatty acids are anteiso-C15:0 and anteiso-C17:0. On the basis of morphological, physiological and chemotaxonomic characteristics, together with DNA-DNA reassociation values and 16S rRNA gene sequence comparison data, the new genus Mycetocola gen. nov. is proposed for these bacteria in the family Microbacteriaceae and three new species are also proposed: Mycetocola saprophilus sp. nov. (type strain CM-01T = IFO 16274T = MAFF 211324T = NRRL B-24119T), Mycetocola tolaasinivorans sp. nov. (type strain CM-05T = IFO 16277T = MAFF 211325T = NRRL B-24120T) and Mycetocola lacteus sp. nov. (type strain CM-10T = IFO 16278T = MAFF 211326T = NRRL B-24121T). The type species of the genus is Mycetocola saprophilus sp. nov. PMID- 11411719 TI - Implications of alternative classifications and horizontal gene transfer for bacterial taxonomy. AB - Following the publication of the Approved Lists, there has been a tendency to regard all subsequent revisions of classification as providing improved nomenclature, to be accepted without question. This takes no account of the fact that such revisions may be based on one of three alternative concepts, phenetic, phylogenetic or polyphasic classification, sometimes leading to different, valid, but incompatible nomenclature, or that some investigations are based only on subsets of relevant taxa and on limited data, leading to incomplete and sometimes confusing revisions of nomenclature. The polyphasic approach to classification has widespread support, although there appears to be a tendency to allow comparative sequence analyses of 16S rDNA to determine classification contrary to the indications of other data. In some cases, classification is based solely on 16S rDNA data. Examples are considered. Consideration is given to the criteria by which taxa are circumscribed, particularly at the level of genus and species. It is suggested that there is a need for reconciliation of the criteria by which taxa at these levels are circumscribed. Recent studies demonstrating the widespread occurrence of horizontal gene transfer suggest that there is a need for caution in monophyletic interpretations, especially when these are based on the analysis of single sequences. PMID- 11411720 TI - Treponema parvum sp. nov., a small, glucoronic or galacturonic acid-dependent oral spirochaete from lesions of human periodontitis and acute necrotizing ulcerative gingivitis. AB - Small oral spirochaetes with a strict dependence on either glucuronic acid (GluA) or galacturonic acid (GalA) were isolated from European patients with periodontitis and from Chinese patients with either gingivitis or acute necrotizing ulcerative gingivitis (ANUG). Thirteen such isolates were similar phenotypically to Treponema pectinovorum ATCC 33768T and this classification was confirmed by 16S rRNA sequencing. However, four isolates differed from T. pectinovorum by their small cell size, by a prominent beta-glucuronidase activity, by a distinct protein and antigen profile, by an inability to grow on pectin as sole source of carbohydrate and by a markedly enhanced growth rate when supplied with a second carbohydrate (L-arabinose, D-galactose, D-glucose, D fructose, D-mannitol, D-mannose, pectin, D-ribose or D-xylose) in addition to the essential GluA/GalA. By 16S rRNA sequencing these four isolates clustered in the recently described phylotype 'Smibert-2'. T. pectinovorum (14 strains) and 'Smibert-2' (four isolates with beta-glucuronidase activity) could each be subdivided into two serotypes based on immunoblot reactivity with two mAbs. Representatives of the two groups, including T. pectinovorum ATCC 33768T, showed a 1:2:1-type periplasmic flagellar arrangement. 'Smibert-2' is described as a novel species, Treponema parvum sp. nov., with isolate OMZ 833T (= ATCC 700770T) proposed as the type strain and OMZ 842 (= ATCC 700773) as reference strain for a second serotype. PMID- 11411721 TI - Clostridium felsineum and Clostridium acetobutylicum are two distinct species that are phylogenetically closely related. AB - The gene sequences encoding the 16S rRNA of Clostridium felsineum DSM 794T and NCIMB 10690T were determined. Both sequences exhibited a relatively very low degree of similarity to the previously determined 16S rRNA gene sequence from C. felsineum DSM 794T. C. felsineum is a member of the major Clostridium cluster, cluster I, and is phylogenetically closely related to Clostridium acetobutylicum. DNA-DNA hybridization results clearly indicated that C. felsineum and C. acetobutylicum belong to distinct species. PMID- 11411722 TI - Lactobacillus psittaci sp. nov., isolated from a hyacinth macaw (Anodorhynchus hyacinthinus). AB - A Gram-positive, non-spore-forming, catalase-negative, facultatively anaerobic coccibacillus to rod-shaped bacterium isolated from a parrot was characterized using phenotypic and molecular taxonomic methods. The unknown bacterium phenotypically resembled lactobacilli and comparative 16S rRNA gene sequencing demonstrated that the organism represents a distinct subline within the Lactobacillus delbrueckii rRNA cluster of the genus. 16S rRNA sequence divergence values of > 6% with recognized Lactobacillus species clearly demonstrated the phylogenetic separateness of the parrot bacterium. On the basis of phylogenetic evidence and the phenotypic distinctiveness of the unknown bacterium, a new species, Lactobacillus psittaci sp. nov., is proposed. The type strain of Lactobacillus psittaci is CCUG 42378T (= CIP 106492T). PMID- 11411723 TI - Taylorella asinigenitalis sp. nov., a bacterium isolated from the genital tract of male donkeys (Equus asinus). AB - Three bacterial isolates that were phenotypically indistinguishable from Taylorella equigenitalis were obtained from the urethral fossae of three male donkeys (Equus asinus), one located in the state of California and the other two in the state of Kentucky, USA. Based on results of pulsed-field gel electrophoresis, the isolate from California differed from the two Kentucky isolates, which were the same. Mares bred artificially (California) or naturally (Kentucky) did not show signs of disease, even though infection with the organism was established in those bred naturally. Mares and, uncharacteristically, all three jacks produced antibodies that reacted in the complement fixation test utilized to identify mares recently infected with T. equigenitalis. Sequence analysis of DNA encoding the 16S rRNA revealed that the gene sequences of these isolates were virtually identical to each other (>99.8% similarity), but different (97.6% similarity) from those of several confirmed isolates of T. equigenitalis. The 16S rDNA sequences of the latter were 100% identical. DNA-DNA hybridization studies revealed a mean hybridization level of 89% between the donkey isolate from California and the donkey isolate from Kentucky. On the other hand, the mean DNA-DNA hybridization level from the donkey isolates with DNA from a strain of T. equigenitalis was 23%. The DNA G+C composition was 37.8 mol% for the two donkey isolates, as well as the strain of T. equigenitalis used in the hybridization studies. These data support our opinion that micro-organisms isolated from the male donkeys are different from T. equigenitalis and it is proposed that they be considered a new species within the genus Taylorella and named Taylorella asinigenitalis sp. nov. The type strain is strain UCD-1T (= ATCC 700933T = LMG 19572T). PMID- 11411724 TI - Comamonas nitrativorans sp. nov., a novel denitrifier isolated from a denitrifying reactor treating landfill leachate. AB - A group of Gram-negative denitrifying bacteria has been isolated from a denitrifying reactor treating landfill leachate. The new isolates produced both oxidase and catalase and showed growth on acetate, butyrate, n-caproate, i butyrate, i-valerate, propionate, n-valerate, lactate, alanine, benzoate, phenylalanine and ethanol. No growth was observed on sugars. The bacteria could perform anoxic reduction of nitrate, nitrite and nitrous oxide to nitrogen, coupled to the oxidation of the same substrates as those used under aerobic conditions, except for aromatic compounds. They were very efficient denitrifiers, as estimated from the specific rate of N2 gas production. All the strains showed the same 16S rDNA restriction profile and one of them, designated 23310T, was selected for phylogenetic analysis. The organism clustered within the family Comamonadaceae, being related to Comamonas terrigena (95.8% sequence similarity). On the basis of the phylogenetic analysis, physiological characterization and the ability to efficiently reduce nitrate to N2, it is proposed that the bacterium be assigned to a new species, Comamonas nitrativorans. The type strain is 23310T (= DSM 13191T = NCCB 100007T = CCT 7062T). PMID- 11411725 TI - Zobellia galactanovorans gen. nov., sp. nov., a marine species of Flavobacteriaceae isolated from a red alga, and classification of [Cytophaga] uliginosa (ZoBell and Upham 1944) Reichenbach 1989 as Zobellia uliginosa gen. nov., comb. nov. AB - A mesophilic, aerobic, non-flagellated, gliding bacterium, forming yellow colonies and designated DsijT, was isolated from a red alga on the sea-shore of Roscoff, Brittany, France. DsijT was selected for its ability to actively degrade both agars and carrageenans. The Gram-negative cells occurred singly or in pairs as long rods. The temperature range for growth was 13-45 degrees C, with an optimum at 35 degrees C. The pH range for growth at 35 degrees C was from 6.0 to 8.5, with an optimum around pH 7.0. The NaCl concentrations required for growth at 35 degrees C and pH 7.0 ranged from 5 to 60 g l(-1), with an optimum around 25 g l(-1). The G+C content of the genomic DNA was 42-43 mol%. Phylogenetic analysis of 16S rRNA gene sequences indicated that strain DsijT is closely related to [Cytophaga] uliginosa DSM 2061T. Phenotypic features, however, allowed DsijT and [Cytophaga] uliginosa strains to be distinguished on the basis of ten traits (spreading behaviour, assimilation of eight compounds and amylase production). Their total protein profiles were also different and DNA-DNA hybridization experiments confirmed that DsijT constitutes a new species, distinct from [Cytophaga] uliginosa. Based on the phenotypic features and the phylogenetic relationships of the Flavobacteriaceae, a new genus designated Zobellia gen. nov. is proposed to include Zobellia galactanovorans gen. nov., sp. nov., while [Cytophaga] uliginosa becomes Zobellia uliginosa comb. nov. The type strain of Zobellia galactanovorans is DsijT (= DSM 12802T = CIP 106680T). PMID- 11411726 TI - Comamonas denitrificans sp. nov., an efficient denitrifying bacterium isolated from activated sludge. AB - To find a biomarker for denitrification in activated sludge, five denitrifying strains isolated from three wastewater treatment plants were studied. These strains were selected from among 1,500 isolates for their excellent denitrifying properties. They denitrify quickly and have no lag phase when switching from aerobic to anoxic conditions. All strains have the cd1-type of nitrite reductase. The strains are Gram-negative rods and they all grow as filamentous chains when cultivated in liquid solution. The strains differ in colony morphology when grown on nutrient agar. Almost full-length 16S rDNA sequences were determined and phylogenetic analysis revealed that these strains are positioned among members of the genus Comamonas in the beta-subclass of the Proteobacteria. Signature nucleotides and bootstrap percentages were also analysed to verify this position. Strains 110, 123T, 2.99g, 5.38g and P17 were < or = 96.7% similar to known strains, but > or = 99.7% similar to each other, as judged from their 16S rDNA sequences, and grouped tightly together in the phylogenetic tree. Sequence motifs in the 16S rRNA gene were also found, suggesting the monophyletic origin of these strains. Nevertheless, some strains differed from the others, for example strain 110 branches early from the other strains and 5.38g is phenotypically more inert. Therefore, it is proposed that strains 110, 123T, 2.99g and P17 are classified into a new species, Comamonas denitrificans sp. nov., while the taxonomic status of strain 5.38g will have to await the outcome of further studies. The type strain of Comamonas denitrificans is 123T (ATCC 700936T). PMID- 11411727 TI - Laragh's lessons in pathophysiology and clinical pearls for treating hypertension. PMID- 11411728 TI - Left ventricular diastolic function predicts outcome in uncomplicated hypertension. AB - Whereas left ventricular systolic function has been shown to predict outcome in hypertensive patients without clinical evidence of heart failure, the prognostic power of diastolic function has not been examined. We assessed the relation of mean pulmonary capillary wedge pressure as an index of left ventricular diastolic function to mortality and the incidence of cardiovascular events in patients with uncomplicated hypertension at baseline. Invasive hemodynamic measurements were performed in the period 1972 to 1982 in 172 hypertensive patients without evidence of cardiovascular disease, cardiomegaly or heart failure, and their outcome was ascertained in 1994. Age at baseline averaged 37 +/- 12 years, brachial artery pressure was 162 +/- 30/88 +/- 18 mm Hg, and mean pulmonary wedge pressure 6.3 +/- 3.0 mm Hg. During 2675 patient-years of follow-up, 15 patients died and 34 suffered at least one fatal or nonfatal cardiovascular event. Cox regression analysis showed that pulmonary wedge pressure was a significant predictor of total mortality and of cardiovascular events, after control for age and gender (P < .05). Each 1 mm Hg increase in wedge pressure was associated with a 23% increase in the risk of all-cause mortality and a 13% increase in the risk of a cardiovascular event. The prognostic power was independent of mean brachial artery pressure, body mass index, serum cholesterol, electrocardiographic left ventricular hypertrophy, and smoking at baseline. We conclude that mean pulmonary wedge pressure, which is likely to reflect left ventricular diastolic function in the selected patients of the current study, is a significant and independent predictor of mortality and of cardiovascular events in uncomplicated hypertension. PMID- 11411729 TI - Different patterns of silent cerebral infarct in patients with coronary artery disease or hypertension. AB - The aim of the present study was to clarify the differences in the progression and the characteristics of silent cerebral infarcts (SCI) between patients with coronary artery disease (CAD) and hypertensive patients. Silent cerebral infarcts, a powerful prognostic indicator for stroke, are frequently found in patients with CAD and in hypertensives. However, the differences in the characteristics of SCI and related risk factors between CAD and hypertensive patients have not been thoroughly investigated. We evaluated the number of SCI and their distribution using brain magnetic resonance imaging (T1- and T2 weighted images) in 107 patients with CAD (validated by coronary angiography) and 101 hypertensive patients without history of clinical stroke. The prevalence of multiple SCI (three or more infarcts per person) in patients with CAD and with hypertension was significantly higher than in hypertensives without CAD (46% v 21%; P = .001), whereas that of patients with CAD without hypertension was intermediate (31%). The patients with multi- (two- or three-vessel) vessel diseases (VD) had a significantly higher prevalence of multiple SCI than the hypertensives and the no-stenosis or 1-VD group (68.1% in the 3-VD group, 52.0% in the 2-VD group, 26.8% in the 1-VD group, and 21.0% in the no-stenosis group). Multiple logistic regression analysis revealed that in the CAD group, the number of involved coronary arteries was an independent determinant of SCI (P < .005), whereas in the hypertensive group, age was an independent determinant of SCI (P < .005). When we investigated the distribution of SCI, in the CAD group, SCI in the deep perforator territory (the basal ganglia and the thalamus) were independently associated with the number of involved coronary arteries (P < .005), whereas SCI in the white matter were independently associated with age only (P < .005). In conclusion, SCI were more advanced in the patients with multivessel CAD than in the hypertensive patients, and were more common in patients with CAD and hypertension than in those without hypertension. Coronary atherosclerosis was independently and specifically associated with SCI located in the deep perferator territory but not of SCI located in the white matter. The CAD-atherosclerosis and hypertension may be independently involved in the pathologic process of SCI. PMID- 11411730 TI - Renal macrostructure and cortical circulation in hypertension assessed by dynamic computed tomography. AB - The aim of this study was to assess the macrostructure of the kidney and the grade of heterogeneity in renal cortical circulation in the early stages of essential hypertension. The subjects consisted of 84 patients (<50 years old) who underwent dynamic computed tomography (CT) because of various abdominal diseases and who had no serious hemodynamic abnormalities. The volumes of the whole kidney, cortex, and medulla were measured with the program installed in the CT instrument. In dynamic CT under appropriate conditions, the CT number of each pixel (image element) reflects the blood volume in the pixel. The means and standard deviations were calculated from the CT numbers within the renal cortex. The coefficient of variation (CV) of CT numbers was used as the index of the heterogeneity of renal cortical circulation. The volume ratio of cortex/medulla was significantly (P < .01) smaller in the hypertensive patients (0.80 +/- 0.03 (mean +/- SE)) than in the normotensive subjects (0.92 +/- 0.03). The CV was significantly (P < .01) greater in the hypertensives (n = 23, 0.124 +/- 0.008) than in the normotensives (n = 61, 0.106 +/- 0.003). There was a significant correlation (r = -0.391, P < .001) between the CV of CT numbers and the volume ratio of cortex/medulla, and the relationship between the two variables was independent of other variables. These results suggest that the heterogeneity of renal cortical circulation is increased in the early stages of essential hypertension and is related to changes in renal macrostructure. PMID- 11411731 TI - Prognostic value of left ventricular mass and its evolution during treatment in the Bordeaux cohort of hypertensive patients. AB - Left ventricular hypertrophy (LVH) is associated with an increased risk of cardiovascular complications independently of other known risk factors, but so far the predictive value of its evolution under treatment has been studied relatively little. Since 1983 we have followed up a cohort of initially untreated hypertensive patients with echocardiographic measurements of left ventricular mass (LVM). We analyzed the data on 474 patients with more than 5 years of follow up to assess the prognostic value of LVM and its evolution during treatment for high blood pressure. A total of 40 patients were lost to follow-up. The mean follow-up period was 89 +/- 31 months. A cardiovascular complication was recorded in 40 individuals. There was a strong link between increased LVM and the occurrence of complications (P < .001). At least a second determination of LVM was performed in 311 patients, and the last value before the occurrence of any complication (60 +/- 38 months after the initial examination) was retained. In this subgroup, 28 patients presented with a cardiovascular event. There was a trend toward a reduction of the incidence of events in the group with a regression of LVH as compared to the group with persistent LVH, but there was no difference when patients were split into quartiles according to LVM evolution between baseline and follow-up. Thus, the reduction of LVM on treatment was not a good marker of future cardiovascular events and its seems at least premature to say that LVM fulfils all conditions for a surrogate end point in the evaluation of morbidity/ mortality in the hypertensive. PMID- 11411732 TI - Weight reduction and pharmacologic treatment in obese hypertensives. AB - This study was conducted to evaluate the mechanisms of weight loss-induced blood pressure (BP) reduction focusing, in particular, on the contributions of sympathetic nervous system activity, fasting plasma insulin, and leptin to BP levels, and to delineate the additional influence of antihypertensive drug therapy. Each of five groups of obese hypertensives were treated with the long acting calcium channel blocker (CCB) amlodipine, the angiotensin converting enzyme (ACE) inhibitor enalapril with or without a weight reduction program, or a weight reduction program alone. The goal BP was less than 140/90 mm Hg for the pharmacologic treatment groups. The weight reduction program groups with or without pharmacologic treatment were divided into two groups; weight loss groups who succeeded in weight reduction (> or = 10%) and nonweight loss groups who failed in weight reduction (<10%) in the first 6 months. The final dose of CCB and ACE inhibitor were less in the combined pharmacologic and weight loss groups than in the pharmacologic treatment alone groups or in the pharmacologic and nonweight loss groups. In the weight reduction groups regardless of pharmacologic treatment, the percent reductions from baseline in plasma insulin, leptin, and norepinephrine (NE) were greater in the weight loss groups (> or = 10%) than in the nonweight loss groups (<10%). The reductions in plasma NE, insulin, and leptin were significantly greater and earlier in combined pharmacologic and weight loss groups than in the pharmacologic treatment alone groups. In ACE inhibitor groups, the reductions in plasma NE, in insulin, and especially in leptin were greater than the other groups. In the CCB alone group, reductions in insulin and leptin occurred, but there was no change in plasma NE. Reductions in insulin and leptin in CCB groups were less and occurred later than in the ACE inhibitor groups or the weight reduction alone group. These results show that weight loss associated with favorable metabolic improvements and these improvements are amplified when combined with pharmacologic treatment. Therefore, weight loss should be regarded as an essential component of any treatment program for obesity-related hypertension. A novel finding from this study is that ACE inhibition had a striking effect to lower plasma leptin. Suppression of sympathetic activity, insulinemia, and leptinemia appeared to play a role in the BP reduction accompanying weight loss. PMID- 11411733 TI - Biphasic effect of epinephrine on blood glucose during hyperinsulinemia in borderline hypertensive young men. AB - We aimed to study the glycemic response to epinephrine during hyperinsulinemia and infused epinephrine (0.03 microg/kg/min) for 30 min after 90 min of hyperinsulinemic glucose clamp in 14 borderline hypertensive young men. Plasma epinephrine was increased from 0.34 +/- 0.08 to 2.33 +/- 0.33 nmol/L while insulin and glucose infusions were kept constant with consequent changes in blood glucose. Initially (90 to 95 min), there was a decrease in blood glucose (P = .016) that correlated negatively with glucose disposal rate corrected for insulin (r = -0.55, P = .040) and positively with fasting insulin (r = 0.55). Thereafter, there was an increase in blood glucose (95 to 120 min) (P < .001) that persisted during the recovery period (120 to 140 min). The glucose increase (90 to 140 min) correlated positively with fasting insulin (r = 0.55), systolic blood pressure (r = 0.57), delta epinephrine 90 to 120 min (r = 0.59), and baseline epinephrine (r = 0.57). Blood glucose remained unchanged (P = .207) in a saline control group (n = 6) with a significant group X treatment effect versus epinephrine (P = .003). Thus, epinephrine caused a biphasic response in blood glucose during hyperinsulinemia. The initial dip in glucose was more pronounced with higher insulin sensitivity, corresponding to previous observations during mental stress test. The following increment in blood glucose was positively related to insulin, systolic blood pressure, and epinephrine levels. These data suggest that insulin may modify the glycemic response to epinephrine in a potentially favorable direction and indicate some lag time before epinephrine gains effect. Subjects who are insulin sensitive and have low blood pressure and resting epinephrine levels seem to be less prone to hyperglycemia induced by epinephrine. PMID- 11411734 TI - Evaluation of the placebo effect and reproducibility of blood pressure measurement in hypertension. AB - Pharmacologic studies in hypertension often describe blood pressure (BP) reductions in placebo control groups. This placebo effect is currently debated, as it seems to be related to BP measurement methods and as a regression to the mean phenomenon may lead to misinterpretation. Furthermore, data on pulse pressure are lacking. This study was designed to evaluate the placebo effect on BP and to differentiate it from regression to the mean. According to a crossover design, 26 mild-to-moderate hypertensive patients who were treated with placebo or given no treatment were followed-up for 1 month. Clinic and ambulatory BP was assessed at baseline and at the end of each 1-month period. Placebo administration resulted in significant reductions in clinic systolic, diastolic, and mean BP (P < .01), ambulatory 24-h SBP (P < .05), and daytime systolic, diastolic, and mean BP (P < .01, P < .05, P < .01, respectively). No significant differences were noted for pulse pressure and heart rate or between BP values measured at baseline and after 1 month without treatment. Despite a significant correlation between changes in clinic and ambulatory BP, the scatter of individual data suggests that the placebo response observed with one method cannot be systematically extrapolated to the other method. This study conclusively shows the effect of placebo in mild-to-moderate hypertension on both clinic and ambulatory systolic, diastolic, and mean BP, in which it has been shown to differ from the regression to the mean phenomenon. This effect was not observed for pulse pressure or heart rate. PMID- 11411735 TI - Low-dose alpha/beta blockade in the treatment of essential hypertension. AB - Despite the recent emphasis on combination drug therapy for hypertension, little attention has been given to alpha/beta blockade using agents other than labetalol. The purpose of this study was to 1) compare the efficacy of low-dose alpha/beta blockade using doxazosin + betaxolol, versus monotherapy with an angiotensin converting enzyme inhibitor (quinapril) and a diuretic (hydrochlorothiazide [HCTZ]), and 2) assess the efficacy of low-dose doxazosin. In a crossover study, 21 hypertensive subjects were treated for 3 weeks each with HCTZ, 12.5 to 25 mg/day, quinapril, 10 to 40 mg/day, and a combination of doxazosin, 1 to 4 mg + betaxolol, 5 to 10 mg daily. Doses were titrated to achieve a systolic pressure <130 mm Hg, as assessed by self-recorded home measurements. Home blood pressure decreased 11.5/7.5 mm Hg after HCTZ, 12.9/8.8 mm Hg after quinapril, and 21.2/16.5 mm Hg after doxazosin + betaxolol (P < .001/< .001 v HCTZ and P < .002/< .001 v quinapril). The target systolic pressure was achieved by 33%, 43%, and 71% of subjects, respectively (P = .04 v HCTZ, and .03 v quinapril). Among the 8 subjects in whom doxazosin dosage was increased to the maximum of 4 mg, the mean blood pressure achieved at 4 mg did not differ from that achieved at 2 mg (136/87 v 136/88 mm Hg). We conclude that oral alpha/beta blockade is superior to monotherapy with an angiotensin converting enzyme inhibitor or a diuretic and that maximal or near maximal efficacy can be achieved at a 2-mg dose of doxazosin. Low-dose oral alpha/beta blockade merits greater consideration in the drug therapy of essential hypertension. PMID- 11411736 TI - Arterial distensibility and ambulatory blood pressure monitoring in young patients with neurofibromatosis type 1. AB - Vascular disease is an underestimated complication of neurofibromatosis type 1 (NF1). The few studies available on this disease are based on case reports. The purpose of this study was to evaluate the relationship between 24-h systolic blood pressure (SBP) and 24-h heart rate obtained by ambulatory blood pressure monitoring and the carotid femoral pulse wave velocity, a widely used index of arterial distensibility, evaluated with Complior. We studied 64 young NF1 patients and 30 healthy subjects. There was no difference in pulse wave velocity between NF1 patients and healthy subjects. Ten of the NF1 patients showed 24-h SBP or 24-h diastolic blood pressure (DBP) >95th percentile for age and sex. We divided the NF1 group into subgroups: NF1 patients with 24-h SBP and 24-h DBP < or = 95th percentile for age and sex (NF1A group) and NF1 patients with mean SBP or DBP >95th percentile for age and sex (NF1B group). The pulse wave velocity of NF1A and NF1B patients were 6.3 +/- 1 m/sec and 6.4 +/- 1 m/sec, respectively (P = not significant). A significant relationship was found between 24-h SBP, 24-h heart rate, and pulse wave velocity in healthy subjects, but not in all NF1 patients and also between the NF1A and NF1B groups. Distensibility of the central arteries may be altered by various environmental or genetic factors. Thus, genetic determinants may play a role in the response of the large arteries to blood pressure. The recent discovery of neurofibromin in aortic smooth muscle may explain the vascular abnormalities present in NF1 patients. We emphasize the importance of a careful vascular evaluation using a noninvasive method, such as Complior and a periodic ambulatory blood pressure monitoring to detect NF1 patients at high risk of vascular complications. PMID- 11411737 TI - Efficacy of candesartan cilexetil as add-on therapy in hypertensive patients uncontrolled on background therapy: a clinical experience trial. ACTION Study Investigators. AB - A large-scale, 8-week, open-label, clinical experience trial evaluated the efficacy of the angiotensin II receptor (AT1 subtype) blocker candesartan cilexetil (16 to 32 mg once daily) either alone or as add-on therapy in 6465 hypertensive patients. The study population was 52% female and 16% African American with a mean age of 58 years. It included 5,446 patients who had essential hypertension (HBP) and 1,014 patients who had isolated systolic hypertension (ISH). These patients had either untreated or uncontrolled hypertension (systolic blood pressure [SBP] 140 to 179 mm Hg or diastolic blood pressure [DBP] 90 to 109 mm Hg inclusive at baseline) despite a variety of antihypertensive medications including diuretics, calcium antagonists, angiotensin converting enzyme (ACE) inhibitors, and alpha- or beta-blockers, either singly or in combination. The mean baseline blood pressure for the HBP group was 156/97 mm Hg. Candesartan cilexetil as monotherapy (in 51% of HBP patients) reduced mean SBP/DBP by 18.7/ 13.1 mm Hg. As add-on therapy (in 49% of HBP patients) to various background therapies, candesartan cilexetil consistently reduced mean SBP/DBP further, irrespective of the background therapy: diuretics (17.8/11.3 mm Hg), calcium antagonists (16.6/11.2 mm Hg), beta-blockers (16.5/ 10.4 mm Hg), ACE inhibitors (15.3/10.0 mm Hg), alpha-blockers (16.4/10.4 mm Hg). The mean baseline blood pressure for the ISH group was 158/81 mm Hg. Candesartan cilexetil, as monotherapy (in 34% of ISH patients), reduced SBP/DBP by 17.0/4.4 mm Hg. As add-on therapy (in 66% of ISH patients) to various background therapies, candesartan cilexetil consistently reduced mean SBP/DBP further, irrespective of the background therapy: diuretics (17.4/5.1 mm Hg), calcium antagonists (15.6/3.6 mm Hg), beta-blockers (14.0/4.8 mm Hg), ACE inhibitors (13.4/4.3 mm Hg), and alpha-blockers (11.6/4.5 mm Hg). The further blood pressure lowering effects of candesartan cilexetil as add-on therapy were seen regardless of age, sex, and race. Overall, 6.8% of the 6465 patients withdrew because of adverse events, most commonly headache (6.3%) and dizziness (5.0%). Orthostatic hypotension was infrequent; 0.2% with candesartan cilexetil alone, and 0.8% with candesartan cilexetil as add-on therapy. Thus, candesartan cilexetil either alone or as add-on therapy was highly effective for the control of systolic or diastolic hypertension regardless of demographic background when used in typical clinical practice settings. PMID- 11411738 TI - Correction of carotid augmentation index for heart rate in elderly essential hypertensives. ANBP2 Investigators. Australian Comparative Outcome Trial of Angiotensin-Converting Enzyme Inhibitor- and Diuretic-Based Treatment of Hypertension in the Elderly. AB - Carotid augmentation index (AI) is used as a surrogate measure of arterial stiffness. Although arterial stiffness has been shown to either remain unchanged or increase with an increase in heart rate, AI decreases as heart rate increases. This study aimed to quantify this confounding effect of heart rate on AI. We investigated 873 hypertensives, mean age 72 +/- 5 years, 44% men, mean brachial blood pressure 161 +/- 21/82 +/- 11 mm Hg. Carotid artery tonometry with simultaneous continuous wave Doppler measurement of ascending aortic blood flow was performed. AI was calculated from the carotid pressure waveform. Waveforms were decomposed into their forward and backward components and the time to reflection between the maxima of the forward and backward pressure waves was measured. AI showed a stronger (P < .001) association with ejection time (r = 0.48, P < .001) than with heart rate (r = -0.28, P < .001). Although AI is strongly related to the time to reflection (r = -0.51, P < .001), only a weak association was seen between time to reflection and heart rate (r = 0.16, P < .001) or ejection time (r = -0.12, P < .001). Our analysis in an elderly cohort of patients with essential hypertension demonstrates that AI is related to the time to reflection. It also reiterates that AI is confounded by heart rate without any underlying heart rate-dependent change in wave reflection. In population-based studies the confounding effect of heart rate can potentially be corrected. AI remains strongly (r = -0.52) related to time to reflection after correction for the effects of ejection time on AI. PMID- 11411739 TI - Abnormalities in lipid composition of brush-border membranes isolated from renal cortex of spontaneously hypertensive rats. AB - The lipid composition and fluidity in brush-border membrane vesicles (BBMV) isolated from renal cortex of spontaneously hypertensive rats (SHR) and their normotensive control, Wistar Kyoto (WKY) rats have been studied. The activity of Na+-dependent D-glucose transport has been also determined. A significant increase in total phospholipids and free cholesterol were observed in renal BBMVs from SHR, which led to a decrease in the ratio of phospholipid-to-free cholesterol in these hypertensive rats. A reduction in the content of phosphatidylcholine and an increase in the percentage of sphingomyelin were observed in SHR. As a consequence, a diminished ratio of phosphatidylcholine-to sphingomyelin was present in kidneys from SHR. The content of phosphatidylserine and phosphatidylinositol was reduced in SHR. However, the levels of phosphatidylethanolamine, cardiolipin, and lysophosphatidylethanolamine were increased in hypertensive animals. An increase in the level of saturated fatty acids, together with a decrease in the level of unsaturated fatty acids was responsible for the lower ratio of unsaturated-to-saturated fatty acids found in kidneys from SHR. The ratio of linoleic-to-arachidonic acid was increased in SHR when compared to WKY rats, as a result of the observed decrease in the content of arachidonic acid in hypertensive rats. Studies of diphenylhexatriene fluorescence polarization indicated no changes in the fluidity between BBMVs from both experimental rats, which did not correlate to the observed changes in the lipid composition of these membranes. These modifications in brush-border membrane lipid composition were accompanied by changes in the Na+-dependent D-glucose transport through renal BBMVs from SHR. PMID- 11411740 TI - Hypotensive effect of endothelin-1 in nitric oxide-deprived, hypertensive pregnant rats. AB - Both nitric oxide (NO) and endothelin-1 (ET-1) are important mediators in the regulation of vascular tone during pregnancy and preeclampsia. This study was designed to investigate the ET-1-induced hypotensive effect in late pregnant rats (P) and in NO-deprived hypertensive pregnant rats (TP), a model of preeclampsia. From day 13 of pregnancy Wistar rats were fed a control or an N(omega)-nitro-L arginine-enriched diet. On gestational day 20, mean arterial pressure (MAP +/- SEM, in mm Hg) and heart rate (HR) were measured with a carotid catheter in anesthetized rats after a bolus intravenous injection of several agonists and antagonists. After 7 days of chronic NO synthase inhibition, there was a significant increase in MAP (+45 +/- 3.9, P < .01) and 24-h urinary nitrate excretion was significantly decreased (P < .05). ET-1 bolus injection (0.1 nmol/kg) was rapidly followed by a significant decrease in MAP and a slight delayed increase, with no change in HR. The magnitude of the decrease had significantly dropped off in P (-30 +/- 2.2) as compared to that in TP (-46 +/- 5.1) and in virgin rats (-51 +/- 6.3) (P < .05). In P and TP, in vivo depressor effect was also obtained with sarafotoxin S6c, a specific ETB agonist, and blocked by the specific ETB antagonist BQ-788. After inhibition of cyclooxygenase with acetylsalicylic acid, the ET-1-induced hypotension was not modified either in P or in TP. In conclusion, the present data highlight an enhanced ETB receptor mediated hypotensive effect of ET-1 in anesthetized TP as compared to P. The magnitude of the hypotensive effect of ET-1 observed in TP is of the same order as that in virgin rats and neither NO nor vasodilator prostaglandins seem to be involved in TP. The enhanced hypotensive effect of ET-1 could be a beneficial counter-balancing mechanism in this rat model of preeclamptic pathology where an increased sensitivity to vasoconstrictor agents is generally described. PMID- 11411741 TI - Converting enzyme inhibition with captopril abolishes sympathoexcitation to euglycemic hyperinsulinemia in rats. AB - Converting enzyme inhibition and angiotensin II receptor antagonism attenuate elevations in heart rate and plasma norepinephrine in response to insulin, suggesting that integrity of the renin-angiotensin system is necessary for insulin-induced sympathoexcitation. To test this, we infused vehicle (saline) in control experiments, or insulin (40 mU/min) during euglycemic clamp in captopril pretreated (intravenously 2.5 mg/kg, then 1 mg/kg/h) and in nonpretreated urethane-anesthetized Wistar rats while measuring mean arterial pressure, heart rate, and lumbar sympathetic nerve activity. Although euglycemic hyperinsulinemia produced similar blood pressure (BP) increases in insulin-infused rats (change in mean arterial pressure: +9 +/- 3 mm Hg) compared with vehicle-controls (+6 +/- 2 mm Hg), insulin decreased blood pressure (BP) in captopril-pretreated insulin infused rats (-8 +/- 3 mm Hg). Control rats developed mild heart rate increases (change in heart rate: +28 +/- 15 beats/min), contrasting with a marked tachycardia in insulin-infused rats (+82 +/- 13 beats/min) and a bradycardia in captopril-treated insulin-infused rats (-16 +/- 18 beats/min). As with heart rate, insulin-infused rats experienced large increases in lumbar sympathetic nerve activity (+ 127 +/- 29%), whereas small and equivalent elevations were observed in vehicle-treated rats (+39 +/- 24%) and in captopril-pretreated insulininfused rats (+61 +/- 13%). These observations demonstrate an attenuation of insulin-induced sympathetic activation by renin-angiotensin blockade with captopril in Wistar rats, and suggest that the renin-angiotensin system is critical for insulin to exert its sympathoexcitatory effects. PMID- 11411742 TI - Acute infarct of the left parietal lobe in a hypertensive patient. PMID- 11411743 TI - The use of antihypertensive therapy. PMID- 11411744 TI - Amlodipine and long-term renoprotection in hypertensive patients with renal dysfunction. PMID- 11411745 TI - Obesity hypertension: role of leptin and sympathetic nervous system. AB - Obesity may account for as much as 65% to 75% of human essential hypertension in most industrialized countries. Excess renal sodium reabsorption and a hypertensive shift of renal-pressure natriuresis play a key role in mediating obesity hypertension. Sympathetic activation contributes to obesity-induced sodium retention and hypertension because adrenergic blockade or renal denervation markedly attenuates these changes. Recent observations suggest that leptin and its multiple interactions with other neurochemical pathways in the hypothalamus may be a partial link between excess weight gain and increased sympathetic activity. Short-term administration of leptin into the cerebral ventricles increases renal sympathetic activity, and long-term intravenous leptin infusions in nonobese rodents at rates that raise plasma concentrations to the levels found in severe obesity increase arterial pressure and heart rate through adrenergic activation. Also, transgenic mice that overexpress leptin develop hypertension. Acute studies suggest that the renal sympathetic effects of leptin may depend on interactions with other neurochemical pathways in the hypothalamus, including melanocortin-4 receptors. However, it is unclear whether this pathway or others, such as neuropeptide Y, mediate the long-term effects of leptin on blood pressure. In addition, leptin has other actions, such as stimulation of nitric oxide formation and enhancement of insulin sensitivity, which may tend to reduce blood pressure in some conditions. Although the precise role of these complex interactions in human obesity has not been elucidated, this is an important area for further investigation, especially considering the current epidemic of obesity in most industrialized countries. PMID- 11411746 TI - Insulin resistance and cardiovascular disease. AB - Cardiovascular risk factors cluster in obese individuals. Insulin resistance emerges as a common pathogenetic denominator underlying the risk factor cluster. Defects in nonesterified fatty acids metabolism have been implicated in the abnormal lipid and glucose metabolism which characterize the cluster. Other evidence also leads to the adipocyte as an important contributor to the risk factor cluster and cardiovascular complications through effects not only on fatty acids but also on leptin, plasminogen activator inhibitor-1, and angiotensinogen, to name a few. Fatty acids are elevated among abdominally obese individuals, are more resistant to suppression by insulin, and may contribute to hypertension. Fatty acids may affect blood pressure by inhibiting endothelial nitric oxide synthase activity and impairing endothelium-dependent vasodilation. Fatty acids increase alpha1-adrenoceptor-mediated vascular reactivity and enhance the proliferation and migration of cultured vascular smooth-muscle cells. Several effects of fatty acids are mediated through oxidative stress. Fatty acids can also interact with other facets of cluster, including increased angiotensin II, to accentuate oxidative stress. Oxidative stress, in turn, is implicated in the pathogenesis of insulin resistance, hypertension, vascular remodeling, and vascular complications. A clearer delineation of the key reactive oxygen signaling pathways and the impact of various interventions on these pathways could facilitate a rationale approach to antioxidant therapy and improved outcomes among the rapidly growing number of high-risk, insulin-resistant, obese individuals. PMID- 11411747 TI - Arterial pressure control at the onset of type I diabetes: the role of nitric oxide and the renin-angiotensin system. AB - Little is known about how hyperglycemia in diabetes directly affects renal and cardiovascular function. Therefore, we modified the streptozotocin-model of Type I diabetes in rats to enable chronic cardiovascular study at the earliest stages of diabetes, before there was time for development of vascular structural changes. We showed that the onset of diabetic hyperglycemia increased total peripheral resistance, decreased skeletal muscle blood flow, increased thromboxane production, and caused a transient increase in plasma renin activity (PRA). Mean arterial pressure (MAP) also increased, but the amplitude was modest. Moreover, we measured significant increases in glomerular filtration rate (GFR) and renal plasma flow, and also showed that endothelially mediated vasodilation in skeletal muscle was not impaired. We then tested the hypothesis that nitric oxide (NO) was playing an important role in counteracting a pressor response to the onset of diabetes. Our results showed that induction of diabetes in rats with chronic NO synthase inhibition caused a marked and progressive increase in MAP. In addition, PRA increased progressively under those conditions and the increase in GFR was prevented. This suggests that NO may work to keep arterial pressure in control at the onset of hyperglycemia very early in the development of diabetes, possibly by facilitating renal vasodilation and by suppressing activity of the renin-angiotensin system. However, the mechanisms for these interactions and the role of renal vascular resistance and other factors in mediating the hypertensive response remain unknown. PMID- 11411748 TI - Diabetic nephropathy in African Americans. AB - Diabetic nephropathy (DN) is the number one cause of end-stage renal disease in United States and is highly prevalent in African Americans. We have found that among African Americans in Mississippi diabetic nephropathy appears to affect females more than males, which may be related to increased rates of obesity and diabetes in African American women. Glycemic control and control of blood pressure is essential to prolong renal survival and to protect against cardiovascular events. Angiotensin-converting enzyme inhibitors reduce cardiovascular mortality in diabetics and are tolerated in advanced renal disease. The impact of glycemic control, appropriate antihypertensives, and the optimal level of blood pressure control in African Americans with advanced DN require further study. This article reviews the impact, clinical characteristics, risk factors, and treatment of diabetic nephropathy in African Americans. PMID- 11411749 TI - The sympathetic neurobiology of essential hypertension: disparate influences of obesity, stress, and noradrenaline transporter dysfunction? AB - Although the importance of sympathetic nervous activation in the pathogenesis of essential hypertension is well documented, the exact pathophysiology of the sympathetic nervous dysfunction present remains to be delineated. This review details three relatively new findings of disturbed sympathetic neurobiology in hypertension. Adrenaline cotransmission is present in the cardiac sympathetic nerves of patients with essential hypertension, as it is in patients with panic disorder, providing presumptive evidence of exposure to high levels of mental stress in hypertensive patients. In lean patients with hypertension there is also evidence of faulty noradrenaline reuptake into the sympathetic nerves of the heart, an abnormality amplifying the sympathetic neural signal by impairing removal of noradrenaline from the synaptic cleft. If both abnormalities are present in the sympathetic nerves of the kidneys also (which we did not test), there would most probably be a direct contribution to hypertension development. In the kidneys the causal chain between sympathetic overactivity and the development of hypertension is stronger than for the heart. In obesity-related hypertension there is evidence that renal sympathetic tone is high, based on approximately a doubling of the measured rate of spillover of noradrenaline into the renal veins. This increase in sympathetic outflow to the kidneys appears to be a necessary but apparently not a sufficient cause for the development of clinical hypertension, commonly being present also in overweight people with blood pressure in the normotensive range. High renal sympathetic tone in the latter, of course, may well still contribute to elevation of their pressure level, although not on such a scale as to cause clinical hypertension. PMID- 11411750 TI - The sympathetic nervous system and long-term blood pressure regulation. AB - There is considerable evidence that activation of the sympathetic nervous system plays an important role in the pathogenesis of several cardiovascular diseases, including hypertension. However, the mechanisms that account for sympathetic activation and the precise mechanisms that mediate neurally induced hypertension are unclear. In large part, this is due to the difficulty in assessing sympathetic function under chronic conditions. Consequently, acute observations are often extrapolated to infer that similar neural mechanisms are operative under more longterm conditions, an unwarrantable assumption. Nonetheless, considerable theoretical and experimental evidence points to the renal sympathetic nerves as the critical link between the sympathetic nervous system and long-term arterial pressure control. Both chronic increases and decreases in renal adrenergic activity alter renal excretory function and produce sustained elevations and reductions in arterial pressure, respectively. Recent observations, including those in dogs with hemibladders and one denervated kidney, indicate that chronic suppression of renal sympathetic nerve activity and attendant natriuresis are long-term compensatory responses to excess body fluid volumes and hypertension. Furthermore, studies combining deafferentation of cardiac receptors and sinoaortic baroreceptors with the split-bladder preparation suggest that chronic renal sympathoinhibition is mediated by baroreflex mechanisms, an especially important finding given the technical limitations in determining whether baroreflexes completely reset and impact sympathetic activity in chronic hypertension. In contrast to the chronic inhibitory effects of baroreflexes on sympathetic activity, other studies indicate that angiotensin II (Ang II) has sustained renal sympathoexcitatory effects. The opposing long-term effects of baroreflexes and Ang II on renal sympathetic nerve activity support two major hypotheses for sympathetic activation in hypertension: baroreflex dysfunction and activation of the renin-angiotensin system, abnormalities often associated with clinical hypertension. PMID- 11411751 TI - The role of the central nervous system in NaCl-sensitive hypertension in spontaneously hypertensive rats. AB - The central and peripheral nervous system is typically considered to be a short term modifier of sympathetic nervous system activity, but several lines of evidence suggest that they contribute to chronic elevation of arterial pressure in at least some forms of hypertension. Our studies focus on the mechanisms underlying NaCl-sensitive hypertension in the spontaneously hypertensive rat (SHR). When these rats are fed a high NaCl diet, their arterial pressure rapidly increases and is maintained about 30 mm Hg higher than those of pair fed controls. The increase in arterial pressure is associated with a decrease in norepinephrine release, specifically in the anterior hypothalamic nucleus (AHN), resulting in increased sympathetic nervous system activity, peripheral vasoconstriction, and arterial pressure. Furthermore, administration of an alpha2 adrenergic receptor agonist in this area blocks the NaCl-sensitive increase in arterial pressure in the SHR but has no significant effect on arterial pressure in normotensive controls. We have identified three intermediary steps by which dietary NaCl reduces AHN norepinephrine release. First, dietary NaCl causes an increase in plasma NaCl and a blunting of the plasma NaCl circadian rhythm. Second, alterations in plasma NaCl activate osmosensitive neurons in the organum vasculosum of the lamina terminalis (OVLT). Third, OVLT input to the AHN appears to increase the release of atrial natriuretic peptide with a resultant decrease in the local release of norepinephrine. Finally, our evidence demonstrates that these factors lead to an increased rise in sympathetic nervous system activity during the early wake phase in SHR on a high NaCl diet, contributing to NaCl sensitive hypertension in SHR. PMID- 11411752 TI - Functionally specific renal sympathetic nerve fibers: role in cardiovascular regulation. AB - The sympathetic nervous system provides differentiated regulation of the functions of various organs. This differentiated regulation occurs through mechanisms that operate at multiple sites within the classic reflex arc: peripherally at the level of afferent input stimuli to various reflex pathways, centrally at the level of interconnections between various central neuron pools, and peripherally at the level of efferent fibers targeted to various effectors within the organ. In the kidney, increased renal sympathetic nerve activity regulates the functions of the intrarenal effectors: the tubules, the blood vessels, and the juxtaglomerular granular cells. This enables a physiologically appropriate coordination between the circulatory, filtration, reabsorptive, excretory, and renin secretory contributions to overall renal function. Anatomically, each of these effectors has a dual pattern of innervation consisting of a specific and selective innervation by unmyelinated slowly conducting C-type renal sympathetic nerve fibers and an innervation that is shared among all the effectors. This arrangement facilitates maximum flexibility in the coordination of the tubules, the blood vessels, and the juxtaglomerular granular cells so as to produce physiologically appropriate responses to a variety of homeostatic requirements. PMID- 11411753 TI - Role of alpha2-adrenergic receptors in hypertension. AB - This is a brief review of a series of experiments conducted over the past two decades, exploring the role of the alpha2-adrenergic receptors (alpha2-AR) in salt-induced hypertension. The data suggest that salt loading alters the activity of central alpha2-AR, resulting in a hypertensive hyperadrenergic state. Studies to separate the role of each alpha2-AR subtype (alpha2A, alpha2B, and alpha2C) have used genetically engineered mice with disrupted genes for each subtype, or gene treatment in rats with antisense-oligodeoxynucleotides targeting a specific gene sequence. Taken together, the results of these studies indicate that the alpha2A-AR is centrally predominant and exerts a tonic sympathoinhibitory function, whereas peripherally it has a vasoconstrictive effect; the alpha2B-AR is responsible for the central hypertensive sympathoexcitatory response to salt, but is not expressed on vascular wall structures; and the alpha2C-AR seems to have no hemodynamic function. PMID- 11411754 TI - Pathophysiology of pregnancy-induced hypertension. AB - Pregnancy-induced hypertension (PIH) is estimated to affect 7% to 10% of all pregnancies in the United States. Despite being the leading cause of maternal death and a major contributor of maternal and perinatal morbidity, the mechanisms responsible for the pathogenesis of PIH have not yet been fully elucidated. Studies during the past decade, however, have provided a better understanding of the potential mechanisms responsible for the pathogenesis of PIH. The initiating event in PIH appears to be reduced uteroplacental perfusion as a result of abnormal cytotrophoblast invasion of spiral arterioles. Placental ischemia is thought to lead to widespread activation/dysfunction of the maternal vascular endothelium that results in enhanced formation of endothelin and thromboxane, increased vascular sensitivity to angiotensin II, and decreased formation of vasodilators such as nitric oxide and prostacyclin. The quantitative importance of the various endothelial and humoral factors in mediating the reduction in renal hemodynamic and excretory function and elevation in arterial pressure during PIH is still unclear. Investigators are also attempting to elucidate the placental factors that are responsible for mediating activation/dysfunction of the maternal vascular endothelium. Microarray analysis of genes within the ischemic placenta should provide new insights into the link between placental ischemia and hypertension. More effective strategies for the prevention of preeclampsia should be forthcoming once the underlying pathophysiologic mechanisms that are involved in PIH are completely understood. PMID- 11411755 TI - Cardiovascular effects of estrogen. AB - There is a strong link between menopause and increased cardiovascular disease incidence in women, and observational studies suggest that postmenopausal hormone replacement therapy (HRT) reduces cardiovascular disease risk. However, the only published prospective controlled trial of the effects of HRT on cardiovascular outcomes (Heart and Estrogen-Progestin Replacement Study, HERS) showed no net benefit of conventional HRT in women with established coronary disease. An angiograpic study of HRT in a similar patient population showed no regression of established coronary lesions in the active treatment group. Fundamental mechanistic studies of the cellular and molecular events by which hormones protect (or fail to protect) blood vessels are needed to define the role of postmenopausal HRT in cardiovascular disease. Herein, we review studies from our laboratory using the rat carotid injury model showing that estrogen inhibits neointima formation through an estrogen receptor (ER)-dependent mechanism operative in the early period after vascular injury. We have demonstrated that activation of vascular smooth muscle cells and subsequent release of soluble factors including osteopontin stimulate the migration of adventitial fibroblasts in a luminal direction to eventually take up residence in the neointima, and furthermore, that production, release, or posttranslational processing of these factors are inhibited by estrogen through an ER-dependent mechanism. The relevant observational and prospective data on HRT in cardiovascular disease prevention are reviewed, and the cardiovascular effects of estrogen are discussed. PMID- 11411756 TI - Fibrosis and ischemia: the real risks in hypertensive heart disease. AB - The increased cardiovascular morbidity and mortality in hypertension are related to the target organs (ie, heart, brain, kidneys) involvement from vascular disease. Left ventricular hypertrophy (LVH), the major expression of cardiac involvement, is both a structural and functional adaptation to the afterload imposed by the vascular disease. Without this adaptation, cardiac failure would result much earlier in the natural history of hypertensive heart disease (HHD). However, LVH imposes an independent risk that is even greater than the risk associated with the height of systolic or diastolic pressure. The mechanisms that explain this risk have not been defined precisely; several have been postulated. Among these are the following: 1) coronary hemodynamic alterations associated with HHD (ie, increased coronary vascular and minimal vascular resistance, reduced coronary blood flow and flow reserve, and increased blood viscosity); 2) enhanced predisposition for lethal cardiac arrhythmias, cardiac failure, and accelerated atherosclerosis of the coronary arteries (with exacerbation of the ischemia); and 3) collagen deposition and ventricular fibrosis. From the earliest controlled therapeutic trials, deaths from stroke and coronary heart disease were significantly reduced. However, more recent data have indicated that the prevalence of cardiac failure (CHF) continues to rise progressively. The nature of the CHF is no longer primarily from systolic dysfunction, but is now chiefly from diastolic dysfunction. Diastolic dysfunction occurs primarily in the elderly hypertensive patient or in the patient with ischemic heart disease, both of which are associated with increased collagen deposition. Indeed, these effects continue to be suggested by the data from the Framingham Heart Study as well as NHANES-III that indicate CHF is the most common diagnosis occurring in hospitalized patients over 65 years of age. In this report, both experimental and clinical evidence demonstrating that increased ventricular fibrosis occurs in the spontaneously hypertensive rats and in hypertensive patients are provided, and that treatment with the newer antihypertensive agents reduce ventricular hydroxyproline (ie, collagen) content while, at the same time, improve coronary hemodynamics. PMID- 11411757 TI - Aldosterone esters and the heart. AB - There are clinical and experimental situations in which symptoms of mineralocorticoid excess are remediable with mineralocorticoid receptor antagonist treatment, in spite of paradoxically low levels of plasma renin and aldosterone. Several decades ago, a factor isolated from the heart was described that had mineralocorticoid properties like those of aldosterone, but much more potent. It was thought to be similar to aldosterone-18-monoacetate or -21 monoacetate, acetyl derivatives of aldosterone that are very rapidly hydrolyzed in the circulation. In our efforts to confirm and extend these observations, we extracted rat hearts and plasma harvested in a manner that would minimize hydrolysis. The product was subjected to several forms of TLC and HPLC and compared to several acetylated derivatives of aldosterone standards. We found that 68% of the aldosterone extracted from fresh myocardium corresponded to an aldosterone derivative that migrates at the same rate as aldosterone-20 monoacetate. The identity of this compound awaits definitive analysis. Tritiated aldosterone-21-monoacetate hydrolyzed to form aldosterone very rapidly; negligible monoacetate remained in blood left at 37 degrees C for 5 min or in hearts left at room temperature for 30 min. Regulation of aldosterone production serves the requirements of fluid and electrolyte homeostasis provided by transport epithelia, primarily that of the kidney. Nonepithelial actions of aldosterone would be freed of these regulatory constraints if the formation of a more potent derivative of the parent compound to which it is almost immediately hydrolyzed in the circulation were regulated within the nonepithelial target tissues. PMID- 11411758 TI - Are low intakes of calcium and potassium important causes of cardiovascular disease? AB - Inadequate levels of calcium and potassium intake have long been associated with higher blood pressures. Epidemiologic data have suggested these associations and many clinical trials have indicated causal relationships. However, the intervention data are plagued with inconsistent study designs, populations, and results, and there remain many questions regarding dietary recommendations of these nutrients for cardiovascular health. Until recently, nutrition research focused on single-nutrient interventions, generally with disparate results. Recognizing that nutrients are not consumed individually but as combined constituents of a varied diet, efforts in this area have shifted to the role of the overall diet, or dietary patterns, in blood pressure and cardiovascular disease. The suggestions of epidemiologic surveys nearly two decades ago that the total diet has a greater influence on cardiovascular health than do specific components, are now being borne out by randomized controlled trials demonstrating this effect. From these dietary pattern studies, it has become increasingly clear that it is not merely excesses of single nutrients but also deficiencies of multiple nutrients in combination, such as calcium and potassium, that have the greatest dietary effects on cardiovascular health. Several risk factors for cardiovascular disease have now been shown to be reduced with diets that meet the current recommended dietary guidelines, ie, that provide appropriate levels of vitamins, minerals, fiber, and macronutrients. In addition, new data indicate that regular consumption of these diets is associated with decreased mortality. Adequate intake of minerals such as calcium and potassium-specifically derived from foods, where they coexist with other essential nutrients-contributes to cardiovascular as well as overall health. PMID- 11411759 TI - Kidney dysfunction: a sensitive predictor of cardiovascular risk. AB - Changes in renal function can be detected in patients suffering from essential hypertension and heart failure. Detection of an increased serum creatinine, a decreased creatinine clearance and of proteinuria or microalbuminuria in hypertensive patients; and the finding of a diminution in the value of estimated creatinine clearance in heart failure patients are very potent predictors for a worse outcome of the patient. The parameters commonly used to evaluate renal function have to be considered in any stratification of cardiovascular risk in hypertensive and heart failure patients. PMID- 11411760 TI - New approaches to screening for vascular and cardiac risk. AB - Cardiovascular morbid events occur as a consequence of vascular and cardiac disease, which can be identified long before symptoms of organ involvement become apparent. Early detection of abnormalities in the small arteries, especially deficiency of endothelial nitric oxide bioactivity, and of structural changes of remodeling in the left ventricle can identify individuals at risk for morbid events. These individuals should be the target for aggressive lifestyle and pharmacologic interventions known to favorably affect the progression of disease. Techniques are now available for screening of asymptomatic individuals to detect early vascular and cardiac abnormalities likely to lead to progression of disease. Pulse wave analysis provides an assessment of small artery elasticity that appears to correlate with endothelial function. Left ventricular structural alterations can be assessed by ultrasound or by circulating levels of brain natriuretic peptide. The usefulness of these early screening techniques to encourage tailored interventions in susceptible individuals needs to be explored in large-scale trials. PMID- 11411761 TI - Management of hypertensive patients with multiple cardiovascular risk factors. AB - Most hypertensive patients have additional cardiovascular risk factors. Therefore, as stressed in all recent guidelines from expert committees, an evaluation of all risk factors and an assessment of each patient's individual risk status is needed before deciding upon the need for therapy and the type of therapy to be given. In particular, attention must be directed at those risk factors that may be involved in the pathogenesis of hypertension: obesity, excessive sodium intake, physical inactivity, and stress. If active therapy is needed for established hypertension, all modifiable unhealthy lifestyles should be addressed along with appropriate drug therapy. PMID- 11411762 TI - Gene transfer to blood vessels: a research tool and potential therapy. AB - Gene transfer to blood vessels is remarkably effective in altering vasomotor function, and has proven to be a useful tool for studying vascular biology. Gene therapy for cardiovascular diseases is an attractive new approach, which will be used initially for diseases in which pharmacologic approaches are not effective. Preliminary data suggest that two possible targets for gene therapy are prevention of cerebral vasospasm after subarachnoid hemorrhage and treatment of pulmonary hypertension. Perhaps, as better vectors are developed, common clinical problems such as hypertension and hypercholesterolemia also may become targets for gene therapy. PMID- 11411763 TI - Genetic manipulation of the renin-angiotensin system: targeted expression of the renin-angiotensin system in the kidney. AB - The renin-angiotensin system is a classic endocrine system that also exists within individual tissues. All components of the renin-angiotensin system (RAS) are expressed in the kidney suggesting the potential for local production and action of angiotensin II. Although the importance of the kidney in hypertension is unequivocal, our understanding of the functional significance of intrarenal production of angiotensin II remains incomplete. Using genetic manipulation of the mouse genome we generated a novel transgenic model expressing human angiotensinogen specifically in the proximal tubule cells of the kidney. Herein I describe the generation and physiologic characterization of this model, and discuss the implications of our findings in terms of the genetics of human hypertension. The experimental results presented support the hypothesis that a tissue-specific RAS cascade exists in the kidney of this transgenic model, and that this system may play an important role in blood pressure (BP) and renal homeostasis in this model. PMID- 11411764 TI - Development and application of a biological peptide pump for the study of the in vivo actions of angiotensin peptides. AB - In the past few years, a great deal of interest has been focused on the possibility that angiotensin peptides could have direct effects on target tissues independent of their hemodynamic effects. In addition, there has been much speculation on the potential biological roles of angiotensin peptides other than angiotensin II. Unfortunately, a direct test of these possibilities in whole animals has been difficult due to limitations in existing biological systems. In this review, we describe the characteristics of an engineered protein capable of directing the production of a wide variety of peptides to specific tissues and cell types in transgenic animals and discuss its potential applications. PMID- 11411765 TI - Adhesion molecules and their role in vascular disease. AB - A variety of recently discovered glycoproteins have been implicated in cell-cell interactions that are critical for normal hemostasis, immune surveillance, and vascular wall integrity. These cell adhesion molecules (CAM) are known to mediate blood cell (leukocyte, platelet)-endothelial cell interactions that can occur in all segments of the microvasculature under certain physiological (eg, hemostasis) and pathological (eg, inflammation) conditions. The multistep process of leukocyte recruitment illustrates how the coordinated and regulated expression of structurally and functionally distinct families of CAM can elicit a highly reproducible vascular response to inflammation. Selectins mediate the initial, low-affinity leukocyte-endothelial cell interaction that is manifested as leukocyte rolling. This transient binding results in further leukocyte activation and subsequent firm adhesion and transendothelial migration of leukocytes, both of which are mediated by interactions between members of the integrin and immunoglobulin superfamily of CAM. This CAM-regulated process of leukocyte recruitment often results in endothelial cell dysfunction, which can be manifested as either impaired endothelium-dependent vasorelaxation in arterioles, excess fluid filtration in capillaries, and enhanced protein extravasation in venules. Consequently, CAM have been implicated in a variety of vascular disorders (eg, ischemia/reperfusion, atherosclerosis, allograft dysfunction, and vasculitis) and an enhanced expression of these CAM has been invoked to explain the exaggerated microvascular dysfunction associated with some of the risk factors (hypertension, hypercholesterolemia, diabetes) for cardiovascular disease. Monoclonal antibodies and genetically engineered mice have proven to be valuable tools for defining the contribution of CAM to disease progression and provide hope for new diagnostic and therapeutic strategies for cardiovascular diseases. PMID- 11411766 TI - Modulating angiotensin II-induced inflammation by HMG Co-A reductase inhibition. AB - Angiotensin (Ang) II is capable of producing inflammatory changes by signals through its AT1 receptor. Reactive oxygen species production, adhesion molecule expression, chemokines, and other mediators are involved. Nuclear factor-kappaB (NK-kappaB) and activator protein 1 (AP-1) are two of the transcription factors activating the responsible genes. We have studied Ang II-independent modulating effects in a double transgenic rat model harboring the human renin and angiotensinogen genes. We have recently focused on the protective effects of HMG CoA reductase inhibition and review these data here. We found that cerivastatin decreased mortality, lowered blood pressure, preserved renal function, decreased cardiac hypertrophy, and inhibited the entire chain of inflammatory events. Furthermore, NF-kappaB and AP-1 activation was sharply attenuated. We also observed that cerivastatin blocked ERK1/2 phosphorylation in vivo and in vitro. Cerivastatin also inhibited phorbol ester-transmitted events in vascular smooth muscle cells. Because Rho, a member of the Ras protein superfamily is important to Ang II-dependent and -independent vascular smooth muscle signaling events, we suggest that cerivastatin may act by inhibiting the prenylation, membrane anchoring, and subsequent activation of Ras proteins. These data may in part explain cholesterol-independent, HMG-CoA reductase-related, protective effects. PMID- 11411767 TI - Vasoregulatory function of the heme-heme oxygenase-carbon monoxide system. AB - Arterial vessels express one or more heme oxygenase (HO) isoenzymes that catalyze the metabolism of heme to carbon monoxide (CO) and biliverdin. Carbon monoxide promotes vasorelaxation through mechanisms that, depending on the vessels, involve activation of soluble guanylate cyclase, stimulation of calcium-activated potassium channels, or diminished synthesis of constrictor mediators, such as, endothelin and 20-HETE. Inhibitors of HO elicit vasoconstriction in vivo and in isolated pressurized arterioles. Inhibitors of HO also enhance myogenic vasoconstriction, as well as the constriction induced by phenylephrine in several vessels. The blood pressure of awake rats is increased by acute treatment with HO inhibitors, a response that is accompanied by attenuation of baroreflex activity. All in all, it would appear that a product of HO activity manufactured by arterial vessels, presumably CO, promotes vasodilation and decreases the reactivity of vascular smooth muscle to myogenic stimuli and constrictor agonists. In doing so, CO of vascular origin may contribute to the implementation of antihypertensive mechanisms. Carbon monoxide produced in central nervous system structures, for example, the nucleus tractus solitarii, also appears to support a blood pressure-lowering mechanism linked to inhibitory modulation of baroreceptor reflex activity. PMID- 11411768 TI - Role of abnormal nitric oxide systems in salt-sensitive hypertension. AB - A large percentage of human hypertensive patients are salt sensitive, referring to the dependence of hypertension on sodium intake, but the cause of the salt sensitivity is not known. Although several mechanisms may contribute to salt sensitive hypertension, the nitric oxide (NO) system appears to play a major role. Studies in humans and Dahl salt-sensitive (S) rats indicate that NO production is decreased during hypertension. Intravenous L-arginine infusion in Dahl S rats increases NO production and prevents salt-sensitive hypertension. In the Dahl salt-resistant (R) rat, NO production by both inducible NO synthase (iNOS) and neuronal NOS (nNOS) help to prevent salt-sensitive hypertension. Experimental evidence is summarized, indicating that the Dahl S rat has a deficient production of NO by nNOS, although NO production by iNOS appears to moderately decrease salt sensitivity. Other evidence about the importance of NO in salt-sensitive hypertension is reviewed, including the role of the renal NO system. PMID- 11411769 TI - Nitric oxide in the control of renal hemodynamics and excretory function. AB - Experimental evidence has now been amassed to indicate that inhibition of nitric oxide (NO) synthase reduces total or regional renal blood flow by approximately 25 to 30% and markedly increases the renal vascular resistance, demonstrating that basal release of NO helps to maintain the relatively low vascular resistance that is characteristic for the kidney. It has been demonstrated that intraarterial administration of NO synthase inhibitors causes marked reductions in sodium excretion without changes in filtered load and suppressed the arterial pressure-induced natriuretic responses in the kidney. We also demonstrated that a constant rate infusion of a NO donor in dogs pretreated with a NOS inhibitor resulted in increases in sodium excretion but failed to restore the slope of the relation between arterial pressure and sodium excretion, suggesting that an alteration in intrarenal NO production rate during changes in arterial pressure is involved in the mediation of pressure natriuresis. Further experiments in dogs performed in our laboratory have confirmed that there is a direct relationship between changes in arterial pressure and intrarenal NO activity measured using NO sensitive microelectrodes in the renal tissue. These arterial pressure-induced changes in intrarenal NO activity were seen positively correlated with the changes in urinary excretion rates of sodium. Collectively, these data suggest that acute changes in arterial pressure alter intrarenal NO production, which inhibits tubular sodium reabsorption to manifest the phenomenon of pressure natriuresis. PMID- 11411770 TI - Role of endothelin-1 in hypertension and vascular disease. AB - Endothelin-1 (ET-1) is a powerful vasoconstrictor peptide and regulator of blood flow that plays an important role in blood pressure (BP) elevation in some models of experimental hypertension such as DOCA-salt rat, DOCA-salt-treated spontaneously hypertensive rats (SHR), stroke-prone SHR, Dahl salt-sensitive rats, angiotensin II-infused rats, and one-kidney, one-clip Goldblatt rats, but not in SHR, two-kidney, one-clip hypertensive rats, transgenic (mREN2)27 rats, or Nomega-nitro-L-arginine methyl ester chronically treated rats. In those models of hypertension in which ET-1 plays a vasoconstrictor role, ET-1 was shown to be overexpressed in the vessel walls, or BP has been lowered by administration of ET(A/B)- and ET(A)-selective receptor antagonists. In these experimental models, endothelin receptor antagonists also regressed vascular growth and inflammation, and improved endothelial dysfunction. Hypertensive rats treated with endothelin antagonists were protected from stroke and renal injury. In hypertensive rats without generalized vascular overproduction of ET-1, expression of ET-1 was often enhanced in intramyocardial coronary arteries, suggesting a role of ET in myocardial ischemia in hypertension. Moderate-to-severe hypertensive patients presented enhanced expression of pre-proET-1 mRNA in the endothelium of subcutaneous resistance arteries, suggesting that this stage of hypertension may respond particularly well to endothelin antagonism. In some hypertensive patients, exaggerated vascular responses to ET-1 were found. Hypertensive patients with coronary artery disease have increased arterial expression of ET-1. Increased plasma levels of immunoreactive ET have been described in African Americans. ET-1 plays an important role in atherosclerosis, for which hypertension is an important risk factor, and in ischemic heart disease and stroke. Endothelin-1 may also be involved in other forms of vascular disease, including pulmonary hypertension, after angioplasty restenosis, after allograft vasculopathy, and vasculitis. Thus, ET-1 may participate in vascular damage in cardiovascular disease and in BP elevation in experimental models and in human hypertension. Endothelin antagonists could become effective disease-modifying agents in different forms of cardiovascular disease. PMID- 11411771 TI - Abnormal pressure-natriuresis in hypertension: role of cytochrome P450 metabolites of arachidonic acid. AB - The pressure-natriuresis relationship is shifted to higher pressures in genetic and experimental models of hypertension; however, the factors responsible for altering kidney function remain to be determined. In spontaneously hypertensive (SHR) and Lyon hypertensive rats, the resetting of pressure-natriuresis results from increased preglomerular renal vascular tone, whereas sodium reabsorption is elevated in the thick ascending loop of Henle (TALH) of Dahl S rats. Recently, a new route for the renal metabolism of arachidonic acid (AA) has been described, and there is evidence that this pathway contributes to the resetting of renal function in hypertension. In the kidney, cytochrome P450 (CYP) enzymes metabolize AA primarily to 20-HETE and EETs. 20-HETE is a potent constrictor of renal arterioles that has an important role in autoregulation of renal blood flow and tubuloglomerular feedback. 20-HETE and EETS also inhibit sodium reabsorption in the proximal tubule and TALH. In the SHR, the renal production of 20-HETE is elevated and inhibitors of the formation of 20-HETE decrease arterial pressure. Blockade of 20-HETE formation also reduces blood pressure or improves renal function in deoxycorticosterone acetate (DOCA)-salt, angiotensin II--infused, and Lyon hypertensive rats. In contrast, 20-HETE formation is reduced in the TALH of Dahl S rats and this contributes to elevated sodium reabsorption. Induction of 20 HETE synthesis improves pressure-natriuresis and lowers blood pressure in Dahl S rats, whereas inhibitors of the synthesis of 20-HETE promote the development of hypertension in Lewis rats. These findings indicate that the renal production of CYP metabolites of AA is altered in genetic and experimental models of hypertension and that this system contributes to the resetting of pressure natriuresis and the development of hypertension in some models. PMID- 11411772 TI - Role of the angiotensin AT2 receptor in blood pressure regulation and therapeutic implications. AB - The angiotensin (ANG) Type 2 (AT2) receptor is one of two major ANG II receptors that have been identified, cloned, and sequenced. Most of the biologic actions of ANG II are thought to be mediated by the AT1 receptor, but evidence is beginning to emerge that the AT2 receptor has a significant role in the regulation of blood pressure. In the adult rat, the AT2 receptor is expressed, albeit in low concentrations in kidney, mesenteric blood vessels, and heart. Most of the evidence suggests that the AT2 receptor stimulates a vasodilator signaling cascade that includes bradykinin, nitric oxide, and guanosine cyclic 3',5' monophosphate. At lease some of the beneficial actions of AT1 receptor blockade are mediated by the AT2 receptor through this pathway. Several recent studies suggest that AT2 receptors may mediate vasodilation and hypotension. The AT2 receptor represents a potential therapeutic target for agonist action and a candidate molecule in the pathophysiology of hypertension. PMID- 11411773 TI - Psychology publications in the Journal of Sports Sciences 1996-2000: a survey and call to arms. PMID- 11411774 TI - Heart rate and match analysis in pre-pubescent soccer players. AB - The aim of this study was to compare match analysis (using video recordings) and the physiological load (heart rate measured every 5 s, blood lactate measured after the warm-up, first half and second half) of six 11-year-old soccer players during official games of eleven-a-side on a regular-sized pitch (100 x 65 m) and of seven-a-side on a smaller pitch (60 x 40 m). In both games, heart rate exceeded 170 beats x min(-1) 84% of the time, while blood lactate ranged from 1.4 to 8.1 mmol l(-1). No significant differences were recorded for the physiological parameters. For both matches, walking comprised 38% of total time, running 55%, inactivity 3% and jumping 3%. Although there were no significant differences between halves or matches, running for less than 10 s was 10% more frequent in the seven-a-side game. In the seven-a-side game, there were significantly more passes and significantly fewer tackles, suggesting that seven-a-side matches played on smaller pitches may be more suitable for pre-pubescent soccer players. PMID- 11411775 TI - Pre-competitive feeling states and directional anxiety interpretations. AB - In this study, we assessed differences in feeling states indicated by performers who reported being facilitated or debilitated by symptoms associated with competitive anxiety before competition. A sample of high-standard swimmers (n = 190) completed a modified version of the Competitive State Anxiety Inventory-2, including both intensity and direction subscales, and an exploratory checklist of feeling state labels, which comprised positive and negative feeling state labels. Our findings supported the general hypothesis that 'facilitators' report significantly more positive feelings than 'debilitators', who report significantly more negative feelings. Descriptive frequency counts of the largest percentage differences between 'facilitators' and 'debilitators' resulted in the selection of the 'confident' feeling state label on the positive subscales, with it being identified most frequently by the 'facilitators'. Furthermore, of the negative feelings, the groups indicated the label 'anxious' most frequently. This study has extended previous research into the notion of positive and negative anxiety and has revealed individual differences in the combination of feeling states experienced by performers during competition. PMID- 11411776 TI - Empirical links between achievement goal theory and self-determination theory in sport. AB - The aim of this study was to examine the empirical links between achievement goal theory and self-determination theory in sport. Addressing theoretical and methodological limitations of previous research, the study tested the independent and interactive effects of goal orientations and perceived competence on seven motivational variables with different degrees of self-determination. Regression analyses of data collected from 247 British university students showed that task orientation predicted motivational variables with high self-determination. In contrast, ego orientation predicted motivational variables with low self determination. Perceived competence predicted both high self-determined and low self-determined motivational variables. A significant interaction emerged between task and ego orientations in predicting external regulation. The results suggest the adaptive role of task orientation in facilitating self-determined motivation in sport. However, the findings are not conclusive, as the variance explained in most analyses was relatively small. Suggestions are offered for a more comprehensive empirical testing of the links between the two theories. PMID- 11411777 TI - Intermuscular coordination during pendulum rebound exercises. AB - In this study, we assessed coordination during pendulum rebound jumps. To gain insight into the movement coordination strategy, nine experienced male volleyball players performed maximal rebound jumps in a pendulum swing device using three different seat arrangements (90 degrees, 135 degrees and 180 degrees). Two dimensional filming was performed in the right sagittal plane (200 Hz) synchronized with a force platform fitted to the wall (1000 Hz). The surface electromyograms of five muscles were recorded (200 Hz), in conjunction with kinematic and kinetic assessment. During the countermovement phase, the impact forces were attenuated by eccentric contractions of most muscles, which helped to reduce the energy input into the system. The wall reaction forces, net moments and joint power profiles were comparable between conditions. The small differences found between the extreme seat arrangements were attributed to differences in muscle length and the position of the feet. The strategy used during landing was similar to that observed in unconstrained vertical and drop jumps, where the neuromuscular system attenuates the impact forces. During the push-off phase, most muscles were found to contribute to positive work generation, except the semitendinosus, which was stretching throughout the propulsive phase. Despite not being able to exert a large influence over the trunk segment, this muscle was deemed to play an important role in regulating and synchronizing the onset of knee extension, enabling hip extension to occur before extension in the other more distal joints. Our findings show that the neuromuscular system is able to produce consistent movement coordination across experimental conditions and in accordance with the specific task demands and constraints imposed in the movement structure. PMID- 11411778 TI - Use of 'chalk' in rock climbing: sine qua non or myth? AB - Magnesium carbonate, or 'chalk', is used by rock climbers to dry their hands to increase the coefficient of friction, thereby improving the grip of the holds. To date, no scientific research supports this practice; indeed, some evidence suggests that magnesium carbonate could decrease the coefficient of friction. Fifteen participants were asked to apply a force with the tip of their fingers to hold a flattened rock (normal force), while a tangential force pulled the rock away. The coefficient of friction--that is, the ratio between the tangential force (pulling the rock) and the normal force (applied by the participants)--was calculated. Coating (chalk vs no chalk), dampness (water vs no water) and rock (sandstone, granite and slate) were manipulated. The results showed that chalk decreased the coefficient of friction. Sandstone was found to be less slippery than granite and slate. Finally, water had no significant effect on the coefficient of friction. The counter-intuitive effect of chalk appears to be caused by two independent factors. First, magnesium carbonate dries the skin, decreasing its compliance and hence reducing the coefficient of friction. Secondly, magnesium carbonate creates a slippery granular layer. We conclude that, to improve the coefficient of friction in rock climbing, an effort should be made to remove all particles of chalk; alternative methods for drying the fingers are preferable. PMID- 11411779 TI - Exercise and mental health: it's just not psychology! AB - Exercise has been suggested as an effective adjunctive treatment for a range of mental health conditions. In this study, we explored the perceptions of exercise held by course directors of doctoral training programmes in clinical psychology in England. Given the exploratory nature of the study, we used a qualitative design incorporating semi-structured interviews. Although most participants held favourable attitudes regarding exercise, this was related more to exercise being seen as a positive lifestyle activity that is worth encouraging, rather than exercise being recommended as an adjunctive treatment for mental health problems. Inductive analysis identified four themes that underpinned such a response: inconsistent positions on the evidential criteria used to evaluate the role of exercise, which masked themes regarding the perceived 'simplicity' of exercise interventions, a practical adherence to a mind-body dichotomy, and the incompatibility of exercise with traditional models of understanding and treating clinical conditions. The barriers we have highlighted will continue to hinder the consideration of exercise as a common therapeutic adjunct at present. Although further research is required to examine the relationship between exercise and mental health, consideration must also be given to how such research should be disseminated to mental health professionals. PMID- 11411780 TI - The development and initial validation of the Exercise Causality Orientations Scale. AB - Deci and Ryan's causality orientations theory suggests that there are individual differences in motivational orientation towards initiating and regulating behaviour. They described three causality orientations: autonomy, control and impersonal. The aim of this paper is to describe the development and concurrent validity of the Exercise Causality Orientations Scale (ECOS), which was designed to measure the strength of these three orientations within exercise. Altogether, 592 working adults aged 35.0 +/- 11.4 years (mean +/- s) completed the ECOS and measures of self-determination, self-consciousness and social desirability. The analysis was conducted in two parts. First, the data were subjected to confirmatory factor analysis using a multi-trait, multi-method framework. The original model resulted in a poor fit to the data. On the basis of its modification indices, three scenarios with ambiguous items were removed successively, resulting in a scale with good psychometric properties. Secondly, Pearson's correlations were conducted between the subscales of the ECOS and those of the questionnaires used for validation. Most of the results supported a priori hypotheses. In conclusion, our results show the ECOS to have good psychometric properties and they provide some support for its concurrent validity. PMID- 11411781 TI - Overt and covert dimensions of antisocial behavior in early childhood. AB - The present study was designed to assess both the prevalence and structure of antisocial behavior in a normative sample of preschoolers. Prevalence estimates suggested that 40% of preschoolers exhibit at least one antisocial behavior each day. Furthermore, 10% of preschoolers exhibit six or more antisocial behaviors each day. Consistent with research based on older children, factor analyses provided support for conceptualizing antisocial behavior in early childhood as consisting of both overt and covert dimensions. While both overt and covert behaviors had acceptable test-retest reliability, only overt behaviors had acceptable interrater reliability. Finally both overt and covert dimensions of antisocial behavior were uniquely related to general measures of conduct problems, hyperactivity, and adult and peer conflict in the classroom setting. Findings are discussed with regard to early assessment and the developmental course of antisocial behavior. PMID- 11411782 TI - Multimethod assessment of psychopathology among DSM-IV subtypes of children with attention-deficit/hyperactivity disorder: self-, parent, and teacher reports. AB - Using data based on self-, parent, and teacher reports, we assessed various aspects of psychopathology in a large sample of control children and those with ADHD. Confirmatory factor analysis was employed to extract response bias from latent constructs of aggression, anxiety, attention problems, depression, conduct disorder, and hyperactivity. These latent constructs were then entered into logistic regression equations to predict membership in control versus ADHD groups, and to discriminate between ADHD subtypes. Results of the regression equations showed that higher levels of attention problems and aggression were the best predictors of membership in the ADHD group relative to controls. Logistic regression also indicated that a higher degree of aggression was the only significant predictor of membership in the ADHD-Combined group compared to the ADHD-Inattentive group. However, when comorbid diagnoses of Oppositional Defiant Disorder and Conduct Disorder were controlled for in the logistic regression, greater hyperactivity rather than aggression was the sole variable with which to distinguish the ADHD-Combined from the ADHD-Inattentive subtype. Results are discussed in the context of the DSM-IV ADHD nosology and the role of instrument and source bias in the diagnosis of ADHD. PMID- 11411783 TI - Symptom properties as a function of ADHD type: an argument for continued study of sluggish cognitive tempo. AB - Inconsistent alertness and orientation (sluggishness, drowsiness, daydreaming) were reported to accompany Attention Deficit Disorder (ADD) without Hyperactivity in DSM-III. Such Sluggish Cognitive Tempo items were tested in the DSM-IV Field Trial for ADHD, but were discarded from the Inattention symptom list because of poor negative predictive power. Using 692 children referred to a pediatric subspecialty clinic for ADHD, Sluggish Tempo items were re-evaluated. When Hyperactivity-Impulsivity was absent (i.e., using only cases of Inattentive Type plus clinic controls), Sluggish Tempo items showed substantially improved utility as symptoms of Inattention. Factor analyses distinguished a Sluggish Tempo factor from an Inattention factor. When DSM-IV ADHD types were compared, Inattentive Type was uniquely elevated on Sluggish Tempo. These findings suggest that (a) Sluggish Tempo items are adequate symptoms for Inattentive Type, or (b) Sluggish Tempo may distinguish two subtypes of Inattentive Type. Either conclusion is incompatible with ADHD nosology in DSM-IV. PMID- 11411784 TI - The ecological validity of delay aversion and response inhibition as measures of impulsivity in AD/HD: a supplement to the NIMH multimodal treatment study of AD/HD. AB - Impulsivity is a primary symptom of the combined type of Attention Deficit/Hyperactivity Disorder (AD/HD). The Stop Signal Paradigm is premised upon a primary deficit in inhibitory control in AD/HD, whereas the Delay Aversion Hypothesis, by contrast, conceptualizes impulsivity in AD/HD, not as an inability to inhibit a response, but rather as a choice to avoid delay. This study compared the ecological validity of the Stop Signal Task (SST) and Choice-Delay Task (C DT) measure of delay aversion, with respect to their relative utility in discriminating AD/HD children from normal control participants, and their correlations with classroom observations and with ratings of impulsivity and other core AD/HD symptoms on the Conners and SNAP-IV checklists. The tasks exhibited modest discriminant validity when used individually and excellent discriminant validity when used in combination. The C-DT correlated with teacher ratings of impulsivity, hyperactivity, and conduct problems, and with observations of gross motor activity, physical aggression, and an AD/HD composite score. The SST correlated with the observations only. These results suggest that delay aversion is associated with a broad range of AD/HD characteristics whereas inhibitory failure seems to tap a more discrete dimension of executive control. PMID- 11411785 TI - The development of selective attention in children with attention deficit hyperactivity disorder. AB - The influence of age on a selective attention task was studied in a sample of children with and without Attention Deficit Hyperactivity Disorder (ADHD). The impact of methylphenidate (MPH) treatment on selective attention was also investigated in the children with ADHD. Two age groups of children with ADHD and two age groups of control children were tested using a timed computer task. The task consisted of identifying visual target stimuli under various distracter conditions. Distracters varied on the basis of modality (i.e., visual, auditory, or both) and task relevance (i.e., meaningful or irrelevant). Reaction times and accuracy were measured. Children with ADHD were less efficient on the selective attention task than were children without ADHD, and older children were more efficient than younger children in both groups. Children without ADHD were influenced more by the nature of distracters than were children with ADHD. For children with ADHD, MPH improved performance overall. PMID- 11411786 TI - The hopelessness theory of depression: a test of the diathesis-stress and causal mediation components in third and seventh grade children. AB - The goal of the current study was to test the diathesis-stress and causal mediation components of the hopelessness theory of depression in third- and seventh-grade children. The procedure involved an initial assessment of depressive symptoms, hopelessness, and the 3 cognitive styles posited as vulnerability factors by hopelessness theory. The procedure also involved a series of 6 weekly follow-up assessments in which depressive symptoms, hopelessness, and the occurrence of negative events were assessed. A depressogenic attributional style interacted with negative events to predict increases in depressive symptoms in seventh-grade children but not in third-grade children. A depressogenic inferential style about consequences interacted with negative events to predict increases in depressive symptoms in both third- and seventh-grade children. Last, a depressogenic inferential style about the self interacted with negative events to predict increases in depressive symptoms in third- and seventh-grade girls but not boys. None of these interactions were mediated by hopelessness. PMID- 11411787 TI - Specificity of information processing styles to depressive symptoms in youth psychiatric inpatients. AB - Although information processing has been widely studied with depressed adults, little emphasis has been placed on the specificity of resultant findings to depression, as opposed to other psychological disorders. Analogously, even less effort has been directed toward examining the information processing styles of depressed children and adolescents. The present study investigated the specificity of information processing styles to depression and anxiety among 58 youth psychiatric inpatients. To assess information processing, we used a self referent encoding task, in which participants were presented with positive and negative adjectives; participants were asked whether these adjectives described them or not, and were then tested on recall of the adjectives. After controlling for age and gender, lower rates of positive adjective endorsement and lower rates of positive adjective recall were found to be associated with depression, but not anxiety. Additionally, negative adjective endorsement was associated with anxiety symptoms. These results suggest specific cognitive features of depressive symptoms. PMID- 11411788 TI - Does DSM-IV Asperger's disorder exist? AB - DSM-IV criteria for autistic and Asperger's disorders were applied to 157 children with clinical diagnoses of autism or Asperger's disorder. All children met the DSM-IV criteria for autistic disorder and none met criteria for Asperger's disorder, including those with normal intelligence and absence of early speech delay. The reason for this was that all children had social impairment and restricted and repetitive behavior and interests (required DSM-IV symptoms for both autistic and Asperger's disorders) and all had a DSM-IV communication impairment (which then qualified them for a diagnosis of autistic disorder and not Asperger's disorder). Communication problems exhibited by all children were impaired conversational speech or repetitive, stereotyped, or idiosyncratic speech (or both), which are DSM-IV criteria for autism. These findings are consistent with those of 5 other studies and indicate that a DSM-IV diagnosis of Asperger's disorder is unlikely or impossible. PMID- 11411789 TI - Ethanol inhibits L-arginine uptake and enhances NO formation in human placenta. AB - The acute effects of ethanol (20-60 mM) on L-arginine uptake and nitric oxide (NO) formation was investigated in human placental cotyledons perfused at constant flow. Ethanol (40 mM) decreased L-[3H]arginine uptake from 27.6 +/- 2.3 to 15.8 +/- 1.3 per cent (P < 0.05) of the injected dose and significantly enhanced NO levels in the perfusate from 0.88 +/- 0.11 to 2.80 +/- 0.39 microM. Ethanol also elicited the constriction of placental vessels. The effects of ethanol (20-60 mM) on L-arginine uptake and endothelial NO synthase (eNOS) activity were also investigated in cultured human umbilical vein endothelial cells (HUVEC). After 60 min of ethanol (40 mM) exposure, basal L-[3H]arginine uptake (4.7 +/- 0.3 pmol/microg protein/min) was inhibited by 60 per cent (P < 0.05). Basal eNOS activity in HUVEC determined under "no flow" (static) conditions was significantly increased (approximately 1.8 fold) by 60 mM ethanol. These data are consistent with a stimulatory effect of ethanol on eNOS activity in both basal and flow-stimulated conditions, which may serve a protective role against its vasoconstrictive acute effect. While acute ethanol administration inhibits L-arginine uptake, the present results do not allow us to speculate on the effects of chronic ethanol exposure on NO formation in the fetoplacental unity. PMID- 11411790 TI - Oxidized glucocorticoids counteract glucocorticoid-induced apoptosis in murine thymocytes in vitro. AB - 11Beta-hydroxyglucocorticoids (HGCs) are known to induce apoptosis in immature T cells. Here we show that 11-oxoglucocorticoids (OGCs), which are oxidized metabolites of HGCs, counteract the apoptosis-inducing effects of HGC in murine thymocytes in vitro. Corticosterone at concentrations ranging from 0.1-100 microM induced apoptosis in thymocytes obtained from C57BL/6J mice aged 4 weeks, as demonstrated by cell staining with anti-phosphatidylserine antibody, a decrease in mitochondrial membrane potential, and DNA fragmentation. Co-culture of the cells with 10-100 microM of OGCs, dehydrocorticosterone, cortisone, and prednisone significantly inhibited thymocyte apoptosis induced by 1 microM corticosterone, (p<0.006). Among the other 6 physiological metabolites of the HGCs we tested, 20alpha-dehydrocortisol also showed considerable inhibitory effect on corticosterone-induced thymocyte apoptosis. Corticosterone-treatment of thymocytes in vitro decreased the number of CD4 and CD8 double positive cells, while co-culturing the cells with dehydrocorticosterone significantly attenuated this corticosterone effect (p<0.0001). Numbers of double-negative cells and single-positive cells were not significantly affected by corticosterone, dehydrocorticosterone, or both together. These results raised the possibility that OGCs and probably other HGC metabolites can regulate apoptotic cell death of immature double-positive thymocytes induced by HGC. PMID- 11411791 TI - Upregulation of the klotho gene expression by thyroid hormone and during adipose differentiation in 3T3-L1 adipocytes. AB - A defect in the klotho gene expression in mice leads to a syndrome resembling human aging. The klotho gene encodes a single membrane protein whose extracellular domain carries homology to beta-glucosidases. However, either its function or regulatory mechanism of the gene expression still remains unknown. In the present study, we investigated the klotho gene expression in 3T3-L1 adipocytes using quantitative reverse transcription-polymerase chain reaction. Both membrane and secreted forms of the klotho gene were expressed in 3T3-L1 preadipocytes. Accompanied with adipose differentiation, not the secreted form but the membrane form was gradually increased. In 3T3-L1 adipocytes, triiodothyronine significantly increased the expression levels of membrane form of the klotho gene. These results suggest that the expression of membrane and secreted forms of klotho transcripts are regulated by different mechanisms and that the klotho gene product may play a role in adipose differentiation. PMID- 11411792 TI - Enhanced serum glucocorticoid levels mediate the reduction of serosal mast cell numbers in diabetic rats. AB - Rats turned diabetic by treatment with alloxan exhibit a significant reduction in serosal mast cell numbersin parallel with decreased insulin levels in the plasma. Our aim was to investigate the putative involvement of endogenous glucocorticoid hormone in this phenomenon. The findings indicated that rats treated with alloxan responded with an increase in levels of serum corticosterone concomitantly with decreased mast cell numbers in the pleural space. We found that either surgical bilateral adrenalectomy or pretreatment with the steroid antagonist RU 486 (20 mg/kg, i.p.) impaired the drop in pleural mast cell counts in alloxinated rats. Administration of insulin (15 U/kg) prevented the increase in corticosterone levels and restored pleural mast cell levels in diabetic animals. In addition, treatment of naive rats with corticosterone (0.5 mg/kg, s.c.) or dexamethasone (0.1 mg/kg, s.c.), for 3 consecutive days, led to a reduction in the number of mast cells recovered from the pleural cavity as noted in diabetic animals. In contrast, insulin reduced serum corticosterone levels and induced a significant elevation in pleural mast cell numbers in naive rats. We conclude that there is a causative relationship between increased levels of glucocorticoids and down regulation of mast cell numbers associated with the diabetic state, both phenomena clearly sensitive to insulin. PMID- 11411793 TI - Influence of 5alpha- and 5beta-reduced progestins on the contractility of isolated human myometrium at term. AB - It is very well known that progesterone induces uterine relaxation on myometrium contractile activity. However, little attention has been paid to the effect induced by its metabolites on human uterine contractility. Therefore, we set out to analyze the potential relaxing effect of some 5alpha- and 5beta-reduced progesterone derivatives on the spontaneous contractility of myometrium from pregnant women. Samples were obtained by caesarian section at 38-40 weeks of pregnancy. Spontaneous uterine contractions were recorded in vitro in the presence of progesterone, or progestins independently, at different non cumulative microM concentrations. The progestins elicited an immediate relaxing effect that was concentration-dependent. With the exception of two 5alpha-reduced progestins (5alpha and 3beta,5alpha), the remaining progestins used in the present study were more potent than progesterone. The potency order with respect to their IC50 values was: 3alpha,5alpha (35 microM) > 5beta (81 microM) > 3beta,5beta (156 microM) > 3alpha,5beta (205 microM) > P4 (225 microM) > 5alpha (19 mM) > 3beta,5alpha (28 mM). When tissues were washed, the contractile activity was recovered. This rapid and reversible relaxing effect was not blocking by antiprogestin RU 486, suggesting that is not through receptor mediated genomic action. The metabolites from progesterone may also determine the pattern of motility, ensuring the necessary quiescent environment to prevent abortion during gestation. PMID- 11411794 TI - Airway hyperresponsiveness to bronchoconstrictor challenge after wood smoke exposure in guinea pigs. AB - Prior airway exposure to wood smoke induces an increase in airway responsiveness to subsequent smoke inhalation in guinea pigs (Life Sci. 63: 1513, 1998; 66: 971, 2000). To further characterize this airway hyperreactivity, we investigated and compared the airway responsiveness to bronchoconstrictor challenge before and 30 min after sham air exposure or wood smoke exposure in anesthetized and artificially ventilated guinea pigs. Various doses of substance P (0.8-6.4 microg/kg), capsaicin (0.2-3.2 microg/kg), prostaglandin F2alpha (30-3000 microg/kg), histamine (1-8 microg/kg), or acetylcholine (5-20 microg/kg) were intravenously injected at 2-min intervals in successively increasing doses to obtain the dose required to provoke a 200% increase in baseline total lung resistance (ED200). Wood smoke exposure significantly lowered the ED200 of substance P, capsaicin, and prostaglandin F2alpha whereas sham air exposure failed to do so. Furthermore, wood smoke exposure did not significantly alter the ED200 of histamine or acetylcholine. Pretreatment with phosphoramidon (2 mg/kg), an inhibitor of the neutral endopeptidase (the major degradation enzyme of substance P), before smoke exposure did not significantly affect the smoke induced reduction in ED200 of substance P. Sectioning both cervical vagi before smoke exposure did not significantly alter the smoke-induced reduction in ED200 of capsaicin or prostaglandin F2alpha. These results suggest that airway exposure to wood smoke acutely produces airway hyperresponsiveness to substance P, capsaicin, and prostaglandin F2alpha, but not to histamine or acetylcholine. Since the combination of phosphoramidon and wood smoke exposure did not result in an additive potentiation of smoke-induced airway hyperresponsiveness to substance P, it is suggested that an inhibition of the degradation enzyme of substance P may contribute to this increase in airway reactivity. Furthermore, vagally mediated bronchoconstriction does not play a vital role in enhanced airway responsiveness to capsaicin or prostaglandin F2alpha. PMID- 11411795 TI - Propofol-induced cytochrome P450 inhibition: an in vitro and in vivo study in rats. AB - Propofol, a widely used anesthetic drug, is known to inhibit cytochrome P450 activities in vitro. The goal of this study was to compare cytochrome P450 activities in vitro and in vivo in presence of propofol. In vitro (liver microsomes and freshly isolated hepatocytes), cytochrome P450 IA2 and IIB1 activities were measured as the production of resorufin from ethoxy- and pentoxyresorufin, respectively, in presence of various concentrations of propofol. In vivo, cytochrome P450 activities were assessed as the production of 13CO2 from 13C-aminopyrine injected intravenously, during acute administration or after chronic treatment. In vitro results confirmed the dose-dependent inhibitory effect of propofol on cytochrome P450 activities, both on liver microsomes and isolated hepatocytes. In vivo, the acute administration of propofol induced a significant decrease of 13C-aminopyrine metabolism. Chronic treatment with propofol induced a significant inhibition of 13C-aminopyrine metabolism only after 2 weeks. No enzyme induction was observed. In conclusion, our results demonstrate that propofol inhibits cytochrome P450 also in vivo. Drug interactions may thus occur during propofol administration. PMID- 11411796 TI - Renal glucose reabsorption during hypertonic glucose infusion in female streptozotocin-induced diabetic rats. AB - Information regarding the renal glucose transport capacity in diabetes mellitus is limited. These data are needed because two weeks following injection of streptozotocin (STZ), mRNA and protein levels of the glucose transporter, GLUT2, are upregulated in the proximal tubule of the rat. Therefore, we measured renal glucose transport and GLUT2 protein levels in female control rats, and in rats one (STZ-1), two (STZ-2), and three weeks (STZ-3) after STZ injection (65 mg kg( 1), i.p.). Progressive amounts of glucose were infused into anesthetized rats via the femoral vein and renal clearances collected. The amount of glucose reabsorbed, factored by the glomerular filtration rate (GFR) was significantly greater in STZ-3 rats compared with all other groups. In addition, the amount of glucose reabsorbed factored by the amount of glucose filtered was decreased in STZ-1 and STZ-2 compared with controls but was increased in STZ-3. By contrast, renal GLUT2 levels were elevated in all the STZ-treated rats. These data suggest that other factors, functioning either in conjunction with or independent of GLUT2, are required to support an elevated renal glucose transport capacity. PMID- 11411797 TI - Insulin gene expression pattern in rat pancreas during the estrous cycle. AB - Sex steroid hormones influence insulin homeostasis and glucose metabolism, estradiol (E2) and progesterone (P4) induce changes in both fasting and postprandial insulinemia in rodents, however, insulin gene expression during estrous cycle is unknown. The aim of the present study was to determine an insulin gene expression pattern during the estrous cycle in the rat. Groups of 6 adult rats in each day of the estrous cycle were used. Serum P4, E2, testosterone (T) and insulin concentrations were determined by radioimmunoassay (RIA). A Northern blot analysis was performed to assess insulin gene expression in pancreatic tissue. We found a marked variation in insulin gene expression during the estrous cycle. The highest insulin expression was observed during the proestrus day. Interestingly, E2 and P4 but not T levels were correlated with changes in insulin mRNA content. The variations in serum insulin during the cycle were correlated with its mRNA content in pancreas. The overall results showed variations in serum insulin and insulin gene expression during estrous cycle of the rat that correlated with circulating E2 and P4 levels. PMID- 11411798 TI - Proliferation and apoptosis in the epithelium of the developing human cornea and conjunctiva. AB - To determine the distribution of proliferating and apoptotic cells in the human cornea during prenatal and early postnatal development, we examined sections of the bulbar conjunctiva, the limbus as well as the central and peripheral cornea between 11 weeks of gestation and 6 months after birth. The objective was to localize dividing cells by proliferating cell nuclear antigen-like immunoreactivity (PCNA-LI) and apoptotic cells by terminal transferase-mediated nick-end labeling (TUNEL). Before the 17th gestational week, PCNA-LI was absent in all 4 regions examined, indicating negligible cell proliferation during early development. After 20 weeks, strong PCNA-labeling was observed in all regions examined suggestive of high proliferative activity not only in the limbus and the bulbar conjunctiva, but also in the central and peripheral cornea. This rise in proliferative activity was followed by a steady decline: after 28 weeks, anti PCNA staining gradually disappeared in the central and peripheral cornea, so that, at 6 months after birth, it was confined to the limbus and the bulbar conjunctiva, resembling the picture described for the adult cornea. TUNEL positive cells were virtually absent in all 4 regions examined before the 38th gestational week. Apoptotic cells only started to appear at 38 weeks; at this stage, they were confined to the bulbar conjunctival epithelium. At 6 months after birth, TUNEL-positive cells were observed in the bulbar conjunctival epithelium and the entire cornea; the limbus, however remained devoid of apoptotic cells throughout the entire prenatal and early postnatal period. The present study for the first time localizes proliferating and apoptotic cells in the epithelium of the developing human cornea. Three stages of development can be distinguished: Minimal proliferation (until 17th week), vigorous proliferation over the entire cornea including the limbus and the bulbar conjunctiva (until 28th week) and gradual decrease in proliferative activity (after 28th week) accompanied by the appearance of apoptotic cells. PMID- 11411799 TI - Ischemic preconditioning: from the first to the second window of protection. AB - In many species one or more brief coronary occlusions limit the injuries which a subsequent ischemia-reperfusion can produce in the myocardium. A similar protection has been observed in the majority of organ systems. A first period or window of protection can lasts up to 3 hours and is followed by a second window of protection (SWOP) which begins about 24 hours after the brief coronary occlusions and lasts about 72 hours. Increase of the release of endogenous agents such as adenosine and nitric oxide (NO) may be responsible for both windows through the activation of a protein-kinase C (PKC) which in turn activates ATP sensitive potassium (K+(ATP)) channels. Nitric oxide is also reported to act directly on K+(ATP) channels. Recently, it has been suggested that the channels involved in the protection are mitochondrial rather than sarcolemmal. In SWOP the origin of NO is attributed to the activity of an inducible NO-synthase. Free oxygen radicals released during preconditioning are likely to take part in the delayed protection through the production of peroxynitrite which activates PKC and through the increase of the activity of antioxidant enzymes such as Mn superoxide-dismutase. The production of heat shock proteins is considered a marker rather than a mechanism of SWOP. PMID- 11411800 TI - Inhibition of peristaltic activity by cannabinoids in the isolated distal colon of mouse. AB - The effects of the cannabinoid receptor agonist Win 55,212-2 and of the competitive cannabinoid receptor antagonist SR 141716A on the electrically-evoked peristalsis of isolated distal colon of mouse were studied. Intraluminal pressure, longitudinal displacement, ejected fluid volume and changes in morphology of external intestinal wall were simultaneously recorded in the pre drug period and in presence of Win 55,212-2 alone or in combination with SR 141716A. In the pre-drug period (control), peristaltic activity was characterised by regular, monophasic waves and the intraluminal content propelled towards anterograde (oro-aboral) direction with a propulsion velocity of 1.25 +/- 0.1 mm x s(-1). Pressure and shortening waves showed a peak amplitude of 2.44 +/- 0.32 kPa and 1.8 +/- 0.72 mm, respectively. The mean amount of fluid volume ejected during each contraction was 80 +/- 12.6 microl. The addition of Win 55,212-2 [10( 7)-10(-4) M] to the organ bath determined a dose-related attenuation of peristaltic activity consequent to the decrease of circular and longitudinal muscle strength. The decrease of contractile activity was followed by dose dependent decrease of the amount of fluid ejected during peristalsis. The effects of Win 55,212-2 [10(-7)-10(-5) M] were prevented by SR 141716A, indicating the presence of cannabinoid CB1 receptors in the mouse distal colon. SR 141716A alone enhanced both tonic and phasic motor activities in the colonic longitudinal smooth muscle, suggesting that CB1 receptor antagonists could act either through antagonising the effect of endogenous CB1 receptor agonist or by an agonist effect on these receptors. The present results further support the hypothesis that cannabinoids perform a neuromodulatory role in various tracts of gastrointestinal system and first demonstrate their action also in the distal colon of rodents. PMID- 11411801 TI - Chronic treatment with amlodipine modulates adrenocortical angiotensin II receptors in spontaneously hypertensive rats. AB - We investigated the effects of long-term treatment with calcium-antagonist, amlodipine, on angiotensin II receptors in the adrenal cortex of spontaneously hypertensive rats (SHR). Seven-week-old male SHR were treated with oral amlodipine (10 mg/kg/day) or vehicle (saline) for four weeks. Age-matched normotensive Wistar-Kyoto (WKY) rats were treated with the vehicle similar to control SHR. Systolic blood pressure (SBP) showed time-dependent increase in SHR but not in WKY rats, while amlodipine treatment significantly reduced the high SBP in SHR. Plasma renin activity was serially increased in SHR, which was further enhanced by amlodipine treatment. But the plasma aldosterone level which was increased in SHR was not changed by amlodipine. Competitive reverse transcriptase-polymerase chain reaction showed that the level of adrenocortical angiotensin II type 1 receptor (AT1R) mRNA progressively decreased in vehicle treated SHR compared to WKY rats and that 4-week course of amlodipine treatment significantly increased AT1R mRNA in SHR to levels comparable to those in WKY rats. Amlodipine treatment reduced the level of adrenocortical angiotensin II type 2 receptor (AT2R) mRNA in SHR from 8 weeks of age. Thus, chronic amlodipine treatment differently modulates both adrenocortical AT1R and AT2R in SHR in a possibly direct manner. PMID- 11411802 TI - FK506, but not cyclosporin A, prevents mitochondrial dysfunction during hypoxia in rat hepatocytes. AB - Hepatic ischemia/reperfusion injury occurs in the clinical situations including liver transplantation. FK506 and cyclosporin A (CsA) are reported to be hepatotrophic agents in addition to being a powerful immunosuppressive agent. Studies were performed to determine whether the drugs influence a mitochondrial dysfunction under the hypoxic conditions in primary culture model of rat hepatocytes. The Anaeropack system was used for cell culture to create a hypoxia. Cells were treated with FK506 or CsA under the normoxic and hypoxic conditions. Hypoxia markedly decreased intracellular adenosine 5'-triphosphate (ATP) contents and the ketone body ratio (KBR, acetoacetate/beta-hydroxybutyrate) in culture medium as compared with normoxia. FK506 prevented the decreases of ATP contents and the KBR. In contrast, CsA had no effect on either ATP contents or the KBR. FK506, but not CsA, increased the KBR under the normoxic conditions. Under the hypoxic conditions, heat shock protein 70 (Hsp70) was detected after reoxygenation. FK506 enhanced the induction of Hsp70, but CsA again had no effect on Hsp70 induction. These results indicate that FK506 protects the hypoxia injury in part by preventing the mitochondrial dysfunction in concert with the enhancement of heat shock response in hepatocytes. PMID- 11411803 TI - Down-regulation of mt1 melatonin receptors in rat ovary following estrogen exposure. AB - In this study, we have demonstrated that 2-[125I]-iodomelatonin binds specifically to rat ovarian granulosa cell (GC) membranes with high affinity (KD=83 pM; Bmax=3.28 fmol/mg protein). Using immunoblot analysis and an anti-mt1 melatonin receptor antibody, we have also detected mt1 melatonin receptors in rat ovary. Because melatonin has been reported to alter the steroidogenic responses of ovarian tissues to gonadotropins, a physiological role for intra-ovarian melatonin may exist. Thus, in order to investigate a possible intra-ovarian role for melatonin, we have used both an in vivo and in vitro model of follicular development. Treatment of immature (day 21) female rats with estradiol (E; 0.2 mg/d x 3 d; subcutaneous) was used to induce follicular growth. Membranes from both untreated (U) and E-treated animals' ovaries contained high-affinity 2 [125I]-iodomelatonin (I-MEL) binding sites (Kd=83 and 23 pM, respectively). Estradiol treatment in vivo caused a significant decrease (P<0.05) in binding of 2-[125I]-iodomelatonin to ovarian membranes with untreated animals' ovaries having a Bmax=3.28 fmol/mg protein vs. estradiol-treated animals' ovaries having a Bmax=0.92 fmol/mg protein. In addition, following Estradiol treatment, mt1 melatonin receptors in rat ovary were down-regulated (approximately 95%) using immunoblot analysis. Granulosa cells isolated from E-treated rats were further matured in vitro with testosterone (T) and the pituitary gonadotropin follicle stimulating hormone (FSH). Granulosa cells were cultured with either T (10 ng/ml) or FSH (5.71 ng ovine FSH-20/ml) alone, or both FSH and T for 48 h. There was no statistically significant specific binding of 2-[125I]-iodomelatonin to GC membranes cultured with T or FSH alone. However, following a 48-h exposure to FSH and T in vitro specific 2-[125I]-iodomelatonin binding occurred with total 2 [125I]-iodomelatonin binding =3.15 [corrected] fmol/mg protein. Therefore, the existence of hormonally-regulated expression of high-affinity melatonin binding sites suggests that melatonin may have an important intra-ovarian physiological role. PMID- 11411804 TI - Methylphenidate sensitization is modulated by valproate. AB - Repeated administration of the stimulant methylphenidate (MPD) produces sensitization to its own effects. Glutamate, dopamine, and GABA have been implicated in the underlying mechanism of sensitization to stimulants such as amphetamine and cocaine. We have investigated effects of the GABAergic agent sodium valproate (VAL) on the locomotor response to MPD. Activities of male Sprague-Dawley rats were continuously recorded by a computerized activity monitoring system for 15 days. We studied the dose effect of valproate 1) at 50, 100, and 200 mg/kg (i.p.) on motor activities, 2) on the acute response of motor activities to 2.5 mg/kg MPD, and 3) on behavioral sensitization to subsequent repeated injections of MPD. Valproate alone did not significantly affect motor activities. All three doses of valproate attenuated the acute locomotor effects of MPD, while only the 50 mg/kg dose blocked the development of sensitization to subsequent administration. Possible mechanisms involving substrates for the effect of GABA agonists on sensitization are discussed. PMID- 11411805 TI - Serotonin transporter in rat platelets. Level of protein expression underlies inherited differences in uptake kinetics. AB - By breeding selection for the extreme values of platelet serotonin level (PSL), two sublines of Wistar-derived rats, with constitutionally high or low PSL and platelet serotonin uptake (PSU), have been developed. Searching for the basis of these differences, we performed quantitative western blot analysis of serotonin transporter (5HTt) in platelet membranes isolated from both rat sublines. A polyclonal anti-5HTt antibody labeled a single, 5HTt-related 94 kDa protein band in platelet membranes, with significantly stronger intensity in membranes from rats that exhibited a high PSL. We conclude that the inherited differences in PSL and PSU in rats, following breeding selection, are determined by the level of 5HTt expression in platelet membranes. PMID- 11411806 TI - In vitro demonstration of a saturable transport system for leptin across the blood-brain barrier. AB - A saturable blood-to-brain transport system for leptin across the blood-brain barrier (BBB) has been observed in vivo. Since the main component of the non fenestrated microvessels of the BBB is the endothelial cell, we established an in vitro culture system of these cerebrovascular cells to study leptin transport and to determine whether the self-inhibition of leptin transport characteristic of a saturable system occurs at this level. The results show that 125I-leptin crossed from the luminal to abluminal side of a monolayer of cerebral microvessel cells significantly faster than the albumin and lactalbumin controls. This transport of 125I-leptin across an in vitro BBB was significantly faster than in the opposite direction and was dose-relatedly inhibited by the addition of unlabeled leptin. Thus, the results establish that the saturable transport system for leptin across the BBB occurs at the level of the endothelial cells of the BBB. PMID- 11411807 TI - Oxidative insult to human red blood cells induced by free radical initiator AAPH and its inhibition by a commercial antioxidant mixture. AB - This study was carried out to investigate sequel of oxidative insult to human erythrocytes induced by a water-soluble radical initiator, 2,2'-azobis (amidinopropane) dihydrochloride (AAPH) and the effect of a commercially available mixed antioxidant (Blackmores, BioAce Excel), containing alpha tocopherol, ascorbic acid, beta-carotene and some herbal extracts (containing grape seed catechins and milk thistle derived silybin), on lipid peroxidation, degradation of membrane proteins and haemolysis. We performed this study in order firstly to clarify aspects of the mechanism of AAPH induced free radical damage in human erythrocytes and secondly to establish in vitro conditions by which the efficacy of mixed antioxidant preparations may fairly and objectively be compared. In the process of oxidation initiated by peroxyl radical, a rapid loss of reduced glutathione occurred in the first 60 min. Formation of thiobarbitric acid-reactive substances indicative of lipid peroxidation increased subsequently and almost reached maximal levels at 180 min before significant apparent degradation of membrane proteins was detected. At this point, a significant haemolysis occurred. This sequence of events is consistent with the idea that haemolysis is a consequence of lipid peroxidation and the degradation of membrane proteins. The mixed commercial antioxidant, which suppressed lipid peroxidation and protected membrane proteins against degradation induced by peroxyl radicals, also effectively delayed AAPH induced haemolysis. The system we describe provides a sound objective basis for the in vitro comparison of the potential efficacy of the hundreds of antioxidant nutritional supplements currently available in the market place. PMID- 11411808 TI - Catecholamines in human saliva. AB - Catecholamines are readily detectable in human saliva but their origin is unclear. Norepinephrine (NE) was stable in saliva stored at 4 degrees for 2 hours but 11 +/- 3% degraded after storage at 25 degrees for 1 hour. We intravenously infused 3H-NE into humans and measured levels of 3H-NE and its metabolites in both saliva and forearm venous plasma (a site whose plasma NE levels reflect both local uptake and release of NE). 3H-NE levels in saliva continued to rise for 1 hour even though forearm plasma levels had plateaued by 5 min. By 65 min into the infusion the ratio of 3H-NE:non-radioactive NE was similar in saliva and forearm venous plasma. The ratio of NE:epinephrine (E) was similar in saliva and forearm venous plasma at all time points. Chewing induced salivation, and at least tripled the amount of NE, E and 3H-NE released into saliva per minute, but decreased their concentration in saliva by as much as one half. Saliva NE level was unaltered after 15 min of standing but was increased by 31% after 1 hour of upright posture. Our data imply that the NE present in human saliva comes from both the bloodstream and from salivary sympathetic nerves. The finding that diffusion of blood NE into saliva takes roughly 1 hour to complete suggests that NE in saliva is a poor index of acute changes in sympathetic activity. PMID- 11411809 TI - Attenuation of morphine dependence and withdrawal in rats by venlafaxine, a serotonin and noradrenaline reuptake inhibitor. AB - The effects of venlafaxine, a novel serotonin and adrenaline reuptake inhibitor, on the morphine withdrawal and activation of morphine conditioned place preference (CPP), were investigated in rats. Our results showed that the most morphine withdrawal signs, including jumping, writhing, shakes, exploring, lacrimation, piloerection, irritability, and diarrhea, were attenuated by pretreatment with 10 or 20 mg/kg venlafaxine. To investigate the effects of venlafaxine on relapse to opiate dependence, the morphine CPP was used and a dopamine D2 antagonist sulpiride was selected as a control drug. The morphine CPP disappeared following a 28-day drug-free period and appeared again after given a single injection of 1 mg/kg morphine. Acute treatment with sulpiride (25 or 50 mg/kg, i.p.) 30 min prior to 1 mg/kg morphine injection significantly blocked the reacquisition of CPP, while venlafaxine (10 or 20 mg/kg, i.p.) did not show significant effect. However, chronic treatment with venlafaxine (5 or 10 mg/kg, i.p. twice, daily, for seven consecutive days) significantly attenuated the reacquisition of morphine CPP, whereas chronic treatment with sulpiride (10 or 20 mg/kg, i.p.) have no significant effect. Our results demonstrated for the first time that venlafaxine strongly attenuates morphine withdrawal and morphine induced reaquisition of PMID- 11411810 TI - Effects of estrogen on the central nervous system. PMID- 11411811 TI - Relationship of mood disturbance to cigarette smoking status among 252 patients with a current mood disorder. AB - BACKGROUND: The relationship between cigarette smoking and mood has received increasing attention. This retrospective study evaluated the relationship between mood disturbance and cigarette smoking status among patients with a current mood disorder. The association between level of nicotine dependence and severity of mood disturbance was also evaluated among current smokers. METHOD: Retrospective data for 252 patients (63.5% male, 85.0% white) admitted for treatment of a mood disorder at the San Diego Veteran Affairs Mental Health Clinical Research Center between November 1988 and June 1997 were studied. All current cigarette smokers at admission (N = 126) were matched with nonsmokers (N = 126) on the primary DSM IV Axis I mood disorder diagnosis, admission status (inpatient or outpatient), gender, age (+/- 5 years), and ethnicity. The Hamilton Rating Scale for Depression (HAM-D), the Beck Depression Inventory, and the Profile of Mood States (POMS) were administered to patients on admission. Conditional logistic regression analysis for matched sets with a backward elimination was used to identify factors independently predictive of current smoking status. RESULTS: A greater number of cups of coffee consumed per day (p = .002), a history of alcoholism (p = .004), and higher POMS fatigue subscale scores (p = .007) were predictive of current smoking status. Among current smokers, the HAM-D terminal insomnia item was positively associated with mean number of cigarettes smoked per day (p = .012). CONCLUSION: Cigarette smoking should be addressed in the treatment of patients with a current mood disorder. Smokers experience greater levels of fatigue than nonsmokers. In addition, higher cigarette consumption levels are associated with mild-to-severe symptoms of terminal insomnia. PMID- 11411812 TI - Sertraline treatment of posttraumatic stress disorder: results of 24 weeks of open-label continuation treatment. AB - BACKGROUND: Posttraumatic stress disorder (PTSD) is typically associated with a high degree of chronicity, comorbidity, and psychosocial disability. The efficacy of sertraline in the acute treatment of PTSD has been confirmed based on the results of 2 large, placebo-controlled studies, but almost no prospective long term treatment studies have been reported. METHOD: One hundred twenty-eight patients who completed 12 weeks of double-blind, placebo-controlled, acute-phase treatment for DSM-III-R-defined PTSD with sertraline were continued into a 24 week open-label continuation phase. Efficacy was evaluated using the endpoint change in the 17-item Clinician Administered PTSD Scale Part 2 (CAPS-2) severity score, the 15-item patient-rated Impact of Event Scale, and the Clinical Global Impressions-Improvement and -Severity of Illness scales as primary outcome measures. Treatment response was defined as > or =30% decrease in the CAPS-2 total severity score (compared with acute-phase baseline score) and a Clinical Global Impressions-Improvement score of 1 or 2. RESULTS: Ninety-two percent of acute-phase responders maintained their response during the full 6 months of continuation treatment. In addition, 54% of acute-phase nonresponders converted to responder status during continuation therapy. Over the 36-week course of acute and continuation therapy, 20% to 25% of the improvement in the CAPS-2 severity score occurred during the continuation phase. Sertraline was well tolerated, with 8.6% of patients discontinuing due to adverse events. A high pretreatment CAPS-2 score (> 75) predicted a longer time to response and a greater likelihood that response occurred after 12 weeks of acute treatment. CONCLUSION: The acute efficacy of sertraline is sustained in the vast majority of patients, and at least half of nonresponders to acute treatment will eventually respond to continued treatment. PMID- 11411813 TI - Estrogen deficiency in severe postpartum depression: successful treatment with sublingual physiologic 17beta-estradiol: a preliminary study. AB - BACKGROUND: The postpartum period is a time when women are vulnerable to depressive disorders, which can be severe and have long-lasting adverse sequelae. In spite of multiple contacts with health care providers, women with postpartum depression often remain unrecognized and untreated. To evaluate the association between estradiol and postpartum depression, we measured serum estradiol concentration and performed an open-label study of physiologic 17beta-estradiol. METHOD: Twenty-three women fulfilling ICD-10 criteria for major depression with postpartum onset were consecutively recruited from a psychiatric emergency unit. Serum estradiol concentrations were measured at baseline and weekly during sublingual 17beta-estradiol treatment for 8 weeks. The treatment effect was assessed using a clinician-rated depression symptom scale, the Montgomery-Asberg Depression Rating Scale (MADRS). RESULTS: At baseline, all patients were severely depressed (mean MADRS total score = 40.7; range, 35-45) and had a low serum estradiol concentration (mean = 79.8 pmol/L; range, 23-140 pmol/L); in 16/23 patients, the concentration was even lower than the threshold value for gonadal failure. During the first week of estradiol treatment, depressive symptoms diminished significantly, resulting in a mean MADRS score of 11.0 (Z = -4.20, p < .001), and serum estradiol concentrations approached those of the follicular phase (mean +/- SD = 342 +/- 141 pmol/L). At the end of the second week of treatment, the MADRS scores were compatible with clinical recovery in 19/23 patients. CONCLUSION: This preliminary study shows that depression symptoms may be rapidly reduced in patients with postpartum depression who have documented estradiol deficiency by treatment with 17beta-estradiol and suggests that estradiol can have significance in the pathophysiology of this condition and may be an option in the treatment of women vulnerable to postpartum depression. PMID- 11411814 TI - Effects of the menstrual cycle on measures of personality in women with premenstrual syndrome: a preliminary study. AB - BACKGROUND: Previous studies suggest that women with premenstrual syndrome (PMS) differ from those without PMS in measures of personality. The purpose of this study was to measure the effect of menstrual cycle phase on personality variables in women with and without PMS. METHOD: The Personality Diagnostic Questionnaire Revised (PDQ-R) was administered in both the follicular and luteal phases to women with PMS (according to National Institute of Mental Health PMS Workshop Diagnostic Guidelines) (N = 40). An asymptomatic control group (N = 20) as well as a symptomatic group of women with DSM-IV-diagnosed recurrent, non-menstrual cycle-related brief depression (N = 20) also completed the questionnaire in both phases. RESULTS: Only women with PMS demonstrated a significant increase in total PDQ-R score (reflecting overall personality disorder) from the follicular to the luteal phase (p < .01). Women with PMS had significantly higher total PDQ-R scores than the asymptomatic controls during both the follicular (p < .05) and luteal (p < .01) phases, whereas there was no significant difference between women with PMS and symptomatic controls during either phase. Subscale scores fit similar patterns, as did the number of women in each group meeting a cutoff score indicative of the presence of personality dysfunction. CONCLUSION: In this preliminary study, women with PMS were unique in demonstrating a menstrual cycle phase effect on PDQ-R score, while their scores in both phases were closer to symptomatic controls than asymptomatic controls. These findings suggest that personality disorder in women with PMS may have both state- and trait-related components. PMID- 11411815 TI - Rapid onset of therapeutic effect of risperidone versus haloperidol in a double blind randomized trial. AB - BACKGROUND: Speed of onset of therapeutic effect is an important dimension of drugs employed to treat psychosis and schizophrenia. Faster onset is desirable to reduce the anguish caused by delusions and hallucinations and to protect patients and others from the consequences of poor judgment associated with psychotic exacerbation. Although sufficient studies have demonstrated that novel antipsychotics have advantages over clinically employed doses of classic drugs in terms of tolerability and aspects of efficacy, less is known about differences in speed of onset of therapeutic effect. This report consists of a post hoc subanalysis of data from a large double-blind, randomized pivotal trial in which we compared onset of therapeutic effect between risperidone and haloperidol. METHOD: During an 8-week period, 227 patients with DSM-III chronic schizophrenia received 4 mg/day of risperidone and 226 patients received 10 mg/day of haloperidol. Symptoms were assessed 6 times (days 0, 7, 14, 28, 42, and 56) using the Positive and Negative Syndrome Scale (PANSS) for schizophrenia and the Clinical Global Impressions-Severity of Illness scale (CGI-S). Data were analyzed using analysis of variance for multiple dependent variables and repeated-measures multivariate analysis of variance. RESULTS: The analyses revealed that patients receiving risperidone improved more rapidly than those receiving haloperidol as measured by PANSS total and CGI-S scores. Differences were most pronounced during the first week of treatment. CONCLUSION: Results suggest that risperidone offers a more rapid response than haloperidol, particularly during the active phase of illness when time to response can be crucial. PMID- 11411816 TI - The apparent effects of ziprasidone on plasma lipids and glucose. AB - BACKGROUND: We examined the effects of ziprasidone on body mass index (BMI) and serum levels of glucose, cholesterol, and triglycerides. METHOD: As part of a multicenter study examining different strategies for switching to ziprasidone from other antipsychotics, we evaluated weight and serum glucose, cholesterol, and triglyceride measurements at baseline and following 6 weeks on ziprasidone treatment in 37 patients at our site. RESULTS: Short-term treatment with ziprasidone appeared to lead to significant reduction in serum cholesterol (p < .001) and triglyceride levels (p = .018) independent of changes in BMI. Ziprasidone treatment appeared to have no significant effect on BMI or glucose level, perhaps due to the small number of subjects. CONCLUSION: Ziprasidone appears to independently lead to a lowering of serum lipid levels. PMID- 11411817 TI - Paroxetine in the treatment of generalized anxiety disorder: results of a placebo controlled, flexible-dosage trial. AB - BACKGROUND: The objective of this randomized, double-blind, placebo-controlled study was to investigate the efficacy and safety of paroxetine in outpatients with generalized anxiety disorder (GAD). METHOD: Male and female outpatients 18 years and older who met DSM-IV criteria for GAD and had baseline scores of at least 20 on the Hamilton Rating Scale for Anxiety (HAM-A) were randomly assigned to treatment with paroxetine (20-50 mg/day) or placebo for 8 weeks. The primary efficacy variable was the mean change from baseline in the total score of the HAM A. Additional key efficacy variables were the change from baseline in the scores of the HAM-A items anxious mood and tension, the anxiety subscale of the Hospital Anxiety and Depression Scale, and the Sheehan Disability Scale (SDS). The proportions of patients fulfilling response and remission criteria at week 8 were also determined. RESULTS: The intent-to-treat population included 324 patients. At week 8, compared with the placebo group (N = 163), the paroxetine group (N = 161) had a significantly greater reduction of GAD symptoms on all of the above mentioned efficacy variables. On the HAM-A anxious mood item, which encompasses the cardinal symptoms of GAD, significantly greater efficacy was observed from week 1 and on the SDS significantly greater improvement was documented in the domain "social life" as early as week 4 for paroxetine compared with placebo. In both the last-observation-carried-forward and completer data sets, significantly greater proportions of paroxetine-treated patients achieved response or remission by week 8. Treatment with paroxetine was well tolerated, and the number and type of adverse events recorded in the paroxetine group correspond to the known safety profile of this medication. CONCLUSION: Paroxetine in doses of 20 to 50 mg once daily is effective in the treatment of patients with GAD. Improvement of core symptoms of GAD occurs early and is associated with significant reduction in disability after only 8 weeks of treatment. PMID- 11411818 TI - Does mirtazapine have a more rapid onset than SSRIs? AB - BACKGROUND: A single study utilizing a cross-sectional analysis of scores on the Hamilton Rating Scale for Depression (HAM-D) suggested that mirtazapine has a more rapid onset than selective serotonin reuptake inhibitors (SSRIs). Analysis based on the HAM-D may favor drugs with sleep-producing effects. The purpose of the present study was to determine if a review of all studies comparing an SSRI with mirtazapine, utilizing persistent improvement as the dependent variable, would suggest that mirtazapine had a more rapid onset than SSRIs. METHOD: All double-blind studies comparing mirtazapine with SSRIs were analyzed. Included in the analysis to determine speed of onset were 298 patients taking mirtazapine and 285 taking an SSRI. Pattern analysis, which has been described and used by other researchers, was employed to study speed of onset. RESULTS: At the end of each of the 3 studies, the total number of responders for each of the drugs did not differ. However, the proportion of responders with onset of persistent improvement in week 1 was greater for mirtazapine (13%, 38/298) than for the SSRIs (6%, 18/285; chi2 = 6.95, df = 1, p = .008). CONCLUSION: These data support the possibility that mirtazapine may have a more rapid onset than SSRIs. This observation should be considered preliminary because of the retrospective nature of the analysis and the absence of a placebo group. PMID- 11411819 TI - Priapism associated with conventional and atypical antipsychotic medications: a review. AB - BACKGROUND: Priapism is a prolonged, usually painful, and persistent penile erection not usually associated with sexual stimuli, resulting from a disturbance in the normal regulatory mechanisms that initiate and maintain penile flaccidity. This infrequent adverse event of antipsychotic medication use requires emergency evaluation and has potentially serious long-term sequelae including erectile dysfunction. Clinicians prescribing antipsychotic medications should be aware of this rare but serious adverse event. METHOD: A computerized search, using the MEDLINE database (1966-summer 2000), located cases of priapism associated with most conventional antipsychotics as well as with clozapine, risperidone, and olanzapine. The search included no restrictions on languages. Keywords included priapism combined with antipsychotic agents and the names of the currently available atypical antipsychotics. Twenty-nine publications were located using these parameters. Additional publications were reviewed for general background on pathophysiology, evaluation, and management. The quality of the evidence reviewed is limited by the observational and uncontrolled nature of case reports, case series. and review articles. RESULTS: Psychotropic-induced priapism is currently believed to be caused by the alpha1-adrenergic antagonism of these medications. Detumescence is sympathetically mediated, and alpha1-adrenergic antagonism (within the corpora cavernosa) inhibits detumescence. The propensity of individual antipsychotics to induce priapism can presumably be estimated on the basis of alpha1adrenergic blockade affinities. Of the conventional antipsychotics, chlorpromazine and thioridazine have the greatest alpha1 adrenergic affinity and have been most frequently reported to be associated with priapism. Of the atypical antipsychotics, risperidone has greater alpha1 adrenergic affinity, although 3 of the 5 currently U.S. Food and Drug Administration (FDA)-approved atypicals have been reported to be associated with priapism. CONCLUSION: Virtually all antipsychotic medications have been reported to rarely cause priapism due to their alpha-adrenergic antagonism. This adverse event should be considered a urologic emergency. Clinicians should be familiar with this infrequent serious adverse event of antipsychotic medications. PMID- 11411820 TI - Characteristics of social phobia among persons with essential tremor. AB - BACKGROUND: Social phobia symptoms have been reported to be common among patients with essential tremor, but characteristics of this comorbidity have not been systematically described. METHOD: Cases with essential tremor (N = 94) and controls without essential tremor (N = 85), ascertained from movement disorder clinic and community samples, were evaluated for social phobia symptoms (using the social phobia module of the Structured Clinical Interview for DSM-IV Axis I Disorders and the Liebowitz Social Anxiety Scale), characteristics of tremor, and associated disability (via videotaped examination, performance test, and disability questionnaire). RESULTS: Lifetime combined prevalence of primary social phobia and clinically significant social phobia symptoms occurring secondary to essential tremor was 32.7% (16/49) among essential tremor patients in the clinic sample. Essential tremor cases with secondary social phobia symptoms reported a markedly later age at onset of clinically significant social phobia symptoms than essential tremor cases with primary social phobia (51.0 vs. 8.8 years). Cases with secondary social phobia also reported greater fear and avoidance of eating, drinking, and writing in public than essential tremor cases with primary social phobia and control subjects with social phobia. Essential tremor cases with secondary social phobia symptoms also demonstrated more severe tremor and tremor-related disability than essential tremor cases with primary social phobia and essential tremor cases without social phobia. Among all essential tremor cases, severity of social phobia symptoms and tremor independently contributed to disability. CONCLUSION: Social phobia appears to occur in a substantial minority of essential tremor patients, and severity of social phobia symptoms is associated with disability, independent of tremor severity. Persons with social phobia symptoms secondary to essential tremor evidence clinical characteristics that differ from those of persons with primary social phobia. Further research is needed to determine the efficacy of treatment of social phobia in essential tremor patients with significant social phobia symptoms. PMID- 11411821 TI - Mitrazapine-associated palinopsia. PMID- 11411822 TI - Type II error and antidepressants. PMID- 11411823 TI - Increase in plasma levels of clozapine and norclozapine after administration of nefazodone. PMID- 11411824 TI - Understanding depression: a long-term, recurring disorder. PMID- 11411825 TI - Medication supervision and adherence of persons with psychotic disorders in residential treatment settings: a pilot study. AB - BACKGROUND: Little is known about risk factors for and predictors of medication nonadherence within residential facilities. This pilot study examined the association between medication adherence and level of supervision and other environmental and clinical variables among patients with schizophrenia and related psychotic disorders living in supported housing. METHOD: A convenience sample of 74 adult residents with schizophrenia and related psychotic disorders (DSM-IV criteria) living in 4 supported housing facilities in New York City were assessed by their treating psychiatrist for medication cessation during the previous month. Demographic characteristics, medications, supervision, global function as measured by the Global Assessment of Functioning (GAF), and substance abuse were also assessed. A priori hypotheses were that regimen complexity would be directly and medication supervision would be inversely related to medication nonadherence. RESULTS: In multivariate models, lack of direct medication supervision, negative medication attitude, and lower GAF score were associated with increased medication nonadherence in the recent past. CONCLUSION: This pilot study suggests that direct supervision of medication is associated with better adherence in residential treatment settings. This finding is relevant for mental health service planners and clinicians working in these settings. PMID- 11411826 TI - Update: newborn screening for sickle cell disease--California, Illinois, and New York, 1998. AB - Sickle cell disease (SCD) is a common single-gene disorder that affects three of every 1000 black newborns and approximately 50,000 persons in the United States. Children affected with SCD are at increased risk for severe morbidity (e.g., severe hemolytic anemia, splenic dysfunction, pain crises, and bacterial infections) and mortality, especially during the first 3 years of life. In 1993, California, Illinois, and New York collectively accounted for approximately 20% of all births to blacks. All three states offer universal newborn screening for hemoglobinopathies. To assess the effectiveness of newborn screening programs for SCD and for receipt of and compliance with early medical interventions (e.g., penicillin prophylaxis and pneumococcal vaccination and other vaccination patterns), a 3-year collaborative follow-up study was conducted from 1995 through 1998 in California, Illinois, and New York. This report summarizes the results of this study, which demonstrate the difficulty in retrospectively finding children who were screened at birth so that data for evaluating program effectiveness can be assessed. PMID- 11411827 TI - Progress toward global dracunculiasis eradication, June 2000. AB - In 1986, an estimated 3 million persons were infected with dracunculiasis (Guinea worm disease) and another 120 million were at risk for infection. That year and in 1991, the World Health Assembly called for the eradication of dracunculiasis, and as a result of the implementation of the Dracunculiasis Eradication Program (DEP), the annual incidence was reduced by approximately 95% by 1995. This report updates the status of the eradication program as of June 2000, which indicates that dracunculiasis has been eliminated from seven of 20 countries where it was endemic in 1995; however, in parts of Africa, particularly Sudan, dracunculiasis remains a serious public health problem. PMID- 11411828 TI - Varicella outbreaks among Mexican adults--Alabama, 2000. AB - On May 19, 2000, a physician in southern Alabama reported seven cases of varicella to the Alabama Department of Public Health (ADPH). All the cases were in previously healthy young adults living in an apartment complex in town A and working in a poultry processing plant in nearby town B. On May 24, ADPH invited CDC to assist in outbreak investigation and control. This report summarizes the investigation. PMID- 11411829 TI - Outbreak of acute respiratory febrile illness among college students--Acapulco, Mexico, March 2001. AB - On March 30, 2001, CDC was notified by Pennsylvania Department of Health (PDH) of an acute respiratory febrile illness in 44 students from two colleges who traveled to Acapulco, Mexico, for spring break vacation during March 3-18. Within 7-14 days of their return from Acapulco, 21 students presented to health-care providers with illness characterized by fever, chills, dry cough, chest pain, and headache. Two students were hospitalized. On the basis of clinical symptoms and chest radiographs that revealed bilateral, nodular patchy infiltrates, acute pulmonary histoplasmosis was the suspected illness. While in Acapulco, most of the students stayed at the Calinda Beach Hotel and participated in group activities at other recreational locations. PMID- 11411830 TI - Prevalence of risk behaviors for HIV infection among adults--United States, 1997. AB - Human immunodeficiency virus (HIV) prevention programs are directed to persons at risk for acquiring and transmitting HIV because of their sexual behaviors or drug use. Effective HIV prevention requires monitoring risk behaviors among persons who are infected, persons who are at highest risk for infection, and the general population. The Behavioral Risk Factor Surveillance System (BRFSS) provides behavioral data at the state level. Because sexual behavior questions are not part of the BRFSS core instrument, in 1997, an optional module was developed and used by 23 states and Puerto Rico. This report summarizes the analysis of these data, which indicates that 11% of respondents had multiple sex partners and 4.2% reported other high-risk behaviors. These findings underscore the continued need for education about behaviors that place persons at risk for HIV infection, promotion of HIV testing among those who engage in these behaviors, and counseling to reduce risk. PMID- 11411831 TI - Human West Nile virus surveillance--Connecticut, New Jersey, and New York, 2000. AB - West Nile virus (WNV), a mosquitoborne arbovirus identified in New York in 1999, has become enzootic in the northeastern United States, affecting humans, birds, horses, and other mammals. Although no human WNV infection was identified in Connecticut or New Jersey in 1999, 62 persons with WNV illness, including seven deaths, were detected in New York City (NYC) and nearby New York counties. In 2000, these jurisdictions implemented active surveillance (AS) and enhanced passive surveillance (EPS) to detect human illness; 21 persons were identified with acute WNV infection (14 in New York, six in New Jersey, and one in Connecticut), including two deaths (one each in New York and New Jersey). This report summarizes the human WNV surveillance systems in Connecticut, New Jersey, New York, and NYC and recommends EPS for hospitalized patients with encephalitis of unknown etiology for the continental United States. PMID- 11411832 TI - Progress toward poliomyelitis and dracunculiasis eradication--Sudan, 1999-2000. AB - Sudan began poliomyelitis and dracunculiasis eradication activities in 1994 and 1995, respectively, in response to resolutions by the World Health Assembly of the World Health Organization (WHO). Sudan poses special obstacles to global eradication campaigns as a result of the disruption caused by ongoing civil war in the vast southern part of the country. The activities of both programs are summarized in this report, which indicated that substantial progress was made to eradicate polio and control of dracunculiasis improved slightly. Continued commitment of resources, access to persons in areas of conflict, and a peaceful resolution of civil unrest are needed to eradicate both diseases. PMID- 11411833 TI - Identification of heterotrophic nanoflagellates by restriction fragment length polymorphism analysis of small subunit ribosomal DNA. AB - Thirty clones derived from twenty isolates of heterotrophic nanoflagellates originating from a variety of marine and freshwater environments were examined by restriction fragment length polymorphism analysis of small subunit ribosomal RNA genes amplified by the polymerase chain reaction (riboprinting). The data were compared with light and electron microscopical identification of the isolates. On morphological criteria, sixteen of the thirty clones belonged to the genus Paraphysomonas De Saedeleer, seven to the genus Spumella Cienkowski, four to the genus Pteridomonas Penard and three to the genus Cafeteria Fenchel and Patterson. Among these taxa, eleven ribotypes were detected by analysis with the restriction enzymes Hinf I, Hae III, Sau3A I, and Msp I. Differentiation of nanoflagellate taxa by the riboprinting method supported taxonomic classification based on morphology at the generic and species level. The utility of the method for discriminating the 'naked' flagellates and for confirming the identity of polymorphic forms among species of Paraphysomonas is demonstrated. PMID- 11411834 TI - Phylogenetic relationships among microsporidia based on rDNA sequence data, with particular reference to fish-infecting Microsporidium balbiani 1884 species. AB - Recently, large discrepancies have been identified between microsporidian systematics based on molecular and traditional characteristics. In the current study the 530f-580r region of the rRNA gene of eight microsporidian species was cloned and sequenced. Included were two unclassified species of Microsporidium Balbiani, 1884 and an unidentified microsporidian that infects the musculature of different sea bream species. Sequence identities in excess of 98% indicated that these three species almost certainly are members of the same genus. Phylogenetic analyses of all microsporidian sequence data available for this region of the gene (20 species) and for partial small subunit sequences (51 species of 21 genera) revealed these species to be distinct from the family Pleistophoridae Doflein, 1901 and closely related them to the genus Sproguea Weissenberg, 1976. This clade was found to comprise a sister taxon to that containing the vast majority of fish-infecting species. Broad cladistic divisions were found between terrestrial insect-infecting and fish-infecting species, which together are distant from the aquatic insect-infecting microsporidia. The rRNA gene of certain fish-infecting genera was found to be more highly conserved than previously reported. This has implications for its utility in diagnostic assays and phylogenetic studies at, or close to, the species level. PMID- 11411835 TI - Development in electrofused conjugants of tetrahymena thermophila. AB - Electric shock can create parabiotic fusions of living Tetrahymena cells. In this study, cells were mated and successful pairs were electrofused with either vegetatively growing cells or other mating pairs. In particular, we electrofused pairs from normal [diploid x diploid] matings with vegetatively dividing cells in G- or M-phase of the cell cycle. We also fused [diploid x diploid] conjugants with mating pairs involving an aneuploid partner [diploid x "star"], which typically undergo an abortive conjugal pathway termed genomic exclusion. Using such parabiotic fusions we identified and characterized two developmentally critical landmarks: 1) the "abort" signal, which is initiated in pairs with nuclear defects (this first becomes evident soon after the completion of Meiosis I or the beginning of Meiosis II); and 2) the "terminal commitment point", a developmental stage in normal [diploid x diploid] pairs after which conjugation no longer responds to a parabiotically transmitted abort signal (this correlates with the onset of the second postzygotic nuclear division). Finally we demonstrate that a conjugal-arrest-activity varies with the vegetative cell cycle, reaching its highest level of activity during M-phase and dropping just after cytokinesis. PMID- 11411836 TI - Redescription of Psilotricha acuminata Stein, 1859 and revisions of the genera Psilotricha and Urospinula (Ciliophora, Hypotrichida). AB - Psilotricha acuminata was described by Stein in 1859 as the type species of the ciliate genus Psilotricha Stein, 1859. The ciliate has rarely been found since, and its infraciliature has never been described with the aid of silver impregnation techniques. We have found P. acuminata Stein, 1859 in soil samples from upland grassland in Scotland (U.K.). Living and healthy organisms of P. acuminata are oblong in outline, and dorso-ventrally compressed. They closely resemble ciliates of the genus Euplotes. The main morphological features used for identification of P. acuminata are the very long and sparse cirri, and the two macronuclei. When the ciliate crawls, the cirri appear stiff and directed backwards. Specimens observed from the ventral side have a protruding anterior end, a rounded or acuminate posterior pole, and a "beak-like" projection to the left side of the posterior end. The ciliate shares characters with the Euplotidae (body shape and reduced ciliature) and with the Oxytrichidae (marginal rows, macronuclei, reduced number of transverse cirri). Because the arrangement of the silver-impregnated infraciliature was unknown, and as the only description of the ciliate was that of Stein (1859a, b), the genus Psilotricha became confused with other hypotrich genera, especially Urospinula Corliss, 1960. Here we provide a full redescription of P. acuminata based on living and silver-impregnated specimens, and a revision of the genera Psilotricha Stein, 1859 and Urospinula Corliss, 1960. We resurrect the genus Urospinula, and give an emended diagnosis for both genera. The species now included within the genus Psilotricha are P. acuminata Stein, 1859 (type species); Psilotricha viridis (Penard, 1922) Kahl, 1932; and Psilotricha geleii (Gelei, 1954) Stiller, 1974. Psilotricha viridis sensu Kahl, 1932 and Psilotricha dragescoi Groliere, 1975 are considered incertae sedis. PMID- 11411837 TI - The nucleariid amoebae: more protists at the animal-fungal boundary. AB - Nucleariid amoebae are naked amoebae, generally characterized by a spherical or sometimes flattened body with radiating filopodia. Most species preferentially consume algal prey or cyanobacteria. Phylogenetic analyses of the small-subunit rRNA coding regions from four nucleariid amoebae place these species near the origin of the animal-fungal divergence, together with the choanoflagellate Corallochytrium and the ichthyosporean clades. The species Nuclearia delicatula, N. moebiusi, and N. simplex form a monophyletic group, while ATCC 30864, tentatively but possibly incorrectly assigned to Nuclearia sp., represents a separate line of descent. These nucleariids are unrelated to the lineage containing the testate filose amoebae (Testaceafilosia). Our findings expand the morphological and phylogenetic diversity of protists at the animal-fungal divergence. PMID- 11411838 TI - Growth and grazing rates of the heterotrophic dinoflagellate Polykrikos kofoidii on red-tide and toxic dinoflagellates. AB - We investigated growth rates, grazing rates, and prey selection of Polykrikos kofoidii when feeding on several species of red-tide and/or toxic dinoflagellates. Polykrikos kofoidii ingested all prey species used in this study, exhibiting positive growth on Lingulodinium polyedrum, Scrippsiella trochoidea, Ceratium furca, Gymnodinium catenatum, Gyrodinium impudicum, Prorocentrum micans, and the toxic dinoflagellate Amphidinium carterae, but not on P. minimum. Specific growth rates of P. kofoidii increased rapidly with increasing density of L. polyedrum, S. trochoidea, C. furca, and G. catenatum before saturating between 500-2,000 ng C ml(-1). Specific growth rates increased continuously when P. kofoidii was fed the other prey species. Maximum specific growth rates of P. kofoidii on G. catenatum (1.12 d(-1)), S. trochoidea (0.97 d( 1)), and L. polyedrum (0.83 d(-1)) were higher than those on C. furca (0.35 d( 1)), A. carterae (0.10 d(-1)), P. micans (0.06 d(-1)), G. impudicum (0.06 d(-1)), and P. minimum (-0.03 d(-1)). Threshold prey concentrations (where net growth = 0) were 54-288 ng C ml(-1). Maximum ingestion and clearance rates of P. kofoidii on these dinoflagellates were 5-24 ng C pseudocolony(-1) d(-1) and 1.0-5.9 microl pseudocolony(-1) h(-1), respectively. Polykrikos kofoidii strongly selected L. polyedrum over S. trochoidea in prey mixtures. Polykrikos kofoidii exhibited higher maximum growth, ingestion, and clearance rates than previously reported for the mixotrophic dinoflagellate Fragilidium cf. mexicanum or the heterotrophic dinoflagellates Protoperidinium cf. divergens and P. crassipes, when grown on the same prey species. Grazing coefficients calculated by combining field data on abundances of Polykrikos spp. and co-occurring red-tide dinoflagellate prey with laboratory data on ingestion rates obtained in the present study suggest that Polykrikos spp. sometimes have a considerable grazing impact on prey populations. PMID- 11411839 TI - Lipid class and fatty acid composition of the protozoan parasite of oysters, Perkinsus marinus cultivated in two different media. AB - The meront stage of the oyster protozoan parasite, Perkinsus marinus, cultivated in two media with different fatty acid profiles was analyzed for its fatty acid and lipid class composition. The composition of fatty acids in the prezoosporangium stage of the parasite as well as that of the host oyster were investigated. Although the lipid class composition of meronts was dominated by phospholipids and triacylglycerol, there was no triaclgycerol detected in either culture medium. Despite the difference in fatty acid composition of the two media, the fatty acid composition of meronts in each medium was dominated by 14:0, 16:0, 18:0, 18:1(n-9), 20: (n-9), 18:2(n-6) and 20:4(n-6), a profile that differed from its host. The quantities of total lipids and fatty acids in meronts increased as the number of meronts increased and far exceeded the initial amounts in the media and in the initial cell inoculum. The meronts harvested 25 d post inoculation, had about 3 to 6 times higher total lipids and 4 to 13 times higher fatty acids than the amounts contained in the media. The fatty acid profiles of both prezoosporangia and oysters resembled each other and consisted primarily of 16:0, 20:4(n-6), 20:5(n-3), 22:2delta7,15, and 22:6(n-3). These results indicate that during meront proliferation, the parasite synthesizes certain fatty acids and lipid classes. For development from meront to prezoosporangium, the parasite may rely on its host for lipid resources. PMID- 11411840 TI - The interaction of protozoa with their potential prey bacteria in the rhizosphere. AB - Extensive colonization of the rhizosphere by beneficial bacteria is desirable but it is not yet clear whether the feeding patterns of protozoa exert any ecological impact on the competence of bacteria in the rhizosphere. To determine whether the successful colonization of the rhizosphere by bacteria can be explained by inherent abilities of the bacteria to resist predation, six bacteria categorized as poor, mediocre or good rhizosphere colonists were tested. The populations of protozoa in the rhizosphere consistently increased to a maximum density in two days but then declined to a stable level. Grazing by protozoa increased the doubling time of all of the test bacteria but did not significantly affect the final densities of bacteria in the rhizosphere. The differences in the colonization ability of the six bacteria tested were not attributable to inherent differences in their susceptibility to predation by protozoa. PMID- 11411841 TI - Diplonema spp. possess spliced leader RNA genes similar to the Kinetoplastida. AB - The phylogenetic placement of the genus Diplonema in relation to fellow phylum members Euglena and Trypanosoma has been uncertain. The spliced leader RNA gene, present in the euglenids and kinetoplastids in distinct forms, was a potential target for resolving this question. The first indication supporting a closer relationship to the kinetoplastids was the recognition of potential spliced leader RNA exon sequences in the genomic DNA of two Diplonema isolates. Examination of total cell RNA revealed transcripts in the anticipated size range at approximately 120 and 130 nt. Specific PCR amplification of a spliced leader RNA gene repeat was performed. The hallmark features of the kinetoplastid-type spliced leader RNA, specifically the 39-nt exon, splice-donor site, Sm-binding site and poly-T tract and the potential to form the requisite stem-loop structures, were found. Diplonema spp. are different from the kinetoplastids by virtue of C residues at positions 4 and 18 in the exon. While the intergenic spacer regions varied in size, each contained the complete sequence or remnants of a 5S ribosomal RNA gene. Possession of a functional spliced leader RNA gene of the kinetoplastid variety in Diplonema supports a closer evolutionary relationship with this group than with the euglenids. PMID- 11411842 TI - The I-antigens of Ichthyophthirius multifiliis are GPI-anchored proteins. AB - The parasitic ciliate Ichthyophthirius multifiliis has abundant surface membrane proteins (i-antigens) that when clustered, trigger rapid, premature exit from the host. Similar antigens are present in free-living ciliates and are GPI-anchored in both Paramecium and Tetrahymena. Although transmembrane signalling through GPI anchored proteins has been well-documented in metazoan cells, comparable phenomena have yet to be described in protists. Since premature exit of Ichthyophthirius is likely to involve a transmembrane signalling event, we sought to determine whether i-antigens are GPI-anchored in these cells as well. Based on their solubility properties in Triton X-114, the i-antigens of Ichthyophthirius are amphiphilic in nature and partition with the detergent phase. Nevertheless, following treatment of detergent lysates with phospholipase C, the same proteins become hydrophilic. Concomitantly, they are recognized by antibodies against a cross-reacting determinant exposed on virtually all GPI-anchored proteins following cleavage with phospholipase C. Finally, when expressed in recombinant form in Tetrahymena thermophila, full-length i-antigens are restricted to the membrane, while those lacking hydrophobic C-termini are secreted from the cell. Taken together, these observations argue strongly that the i-antigens of Ichthyophthirius multifiliis are, in fact, GPI-anchored proteins. PMID- 11411843 TI - Morphology and infraciliature of a marine scuticociliate with a polymorphic life cycle: Urocryptum tortum n. gen., n. comb. AB - A new genus and species combination are proposed for Urocryptum tortum n. gen., n. comb., a scuticociliate with a polymorphic life cycle. This marine ciliate was isolated from a sample taken at Gokasho Bay in Mie Prefecture (Japan). Specimens from different phases of the growth cycle were examined in vivo and with two silver staining techniques. Three life-history stages were observed: an exponential growth phase stage (trophont), a stationary phase stage (tomite), and finally a resting stage (cyst). The exponential growth form is laterally flattened and ovoid; it has 20-24 somatic kineties (SK) and a typical complement of scuticociliate oral structures. Polykinetid 1 (Pk1) has two longitudinal files of 6 kinetosomes (Ks); sometimes one or two additional kinetosomes are located anteriorly. Polykinetid 2 (Pk2) has two files of 6 or 7 Ks; a third file of three widely spaced kinetosomes is located on its right side, as well as a small curved row of 5 Ks positioned on the right hand side of the posterior end of Pk2. Polykinetid 3 (Pk3) has 3 rows of 4, 5, and 7 Ks's perpendicular to the haplokinety (Hk) or paroral membrane. The stationary growth phase cell is spindle shaped and has a similar number of SK that are much more closely spaced than in exponentially growing specimens. Oral infraciliary structures are reduced in size, having fewer kinetosomes and being positioned more anteriorly in the cell. Pk1 is composed of two files of 5-Ks, Pk2 has only two files of 6-7 Ks, and Pk3 has two to three rows of 3-4 Ks. The Hk is displaced anteriorly and becomes straight, losing the shape typical of exponential growth phase cells. Cysts are pyriform with a wide cyst wall; no infraciliary structures were visible. PMID- 11411844 TI - Ultrastructural characteristics of the in vitro cell cycle of the protozoan pathogen of oysters, Perkinsus marinus. AB - Ultrastructural characteristics of vegetative and zoosporangial stages of cultured Perkinsus marinus, a pathogen of the eastern oyster, Crassostrea virginica, were examined by transmission electron microscopy. An axenic cell culture was propagated from infected Chesapeake Bay oyster hemolymph. Different stages of the in vitro cell cycle, including schizonts and different size trophonts, were examined. Trophonts had spherical nuclei with wide perinuclear spaces, mitochondria with tubular cristae, and vacuoles with vacuoplasts. There were micropores on the inside of cell walls. A tubular network in the cytoplasm connected lomasomes to vacuoles, and contained vacuoplast precursor material. Vacuoplasts and precursor material diminished when cell cultures were not fed, suggesting a function in metabolite storage. Cells divided by schizogony or binary fission. Daughter cells in a schizont were not alike, and may specialize for different functions. Some of the daughter cells in a schizont died. Some hypnospores, directly isolated from infected oyster hemolymph enlarged in Ray's fluid thioglycollate medium, and were induced to zoosporulate. Zoosporangia contained varicose, hypha-like structures, whose apical tips gave rise to prezoospores. Ultrastructural characteristics of the vegetative and zoosporangial stages did not resemble any apicomplexan parasites other than members of the genus Perkinsus. PMID- 11411845 TI - Molecular identification of algal endosymbionts in large miliolid foraminifera: 1. Chlorophytes. AB - Large miliolid foraminifers bear various types of algal endosymbionts including chlorophytes, dinoflagellates, rhodophytes, and diatoms. Symbiosis plays a key role in the adaptation of large foraminifera to survival and growth in oligotrophic seas. The identity and diversity of foraminiferal symbionts, however, remain largely unknown. In the present work we use ribosomal DNA (rDNA) sequences to identify chlorophyte endosymbionts in large miliolid foraminifera of the superfamily Soritacea. Partial 18S and complete Internal Transcribed Spacer (ITS) rDNA sequences were obtained from symbionts of eight species representing all genera of extant chlorophyte-bearing Soritacea. Phylogenetic analysis of the sequences confirms the previous fine structure-based identification of these endosymbionts as belonging to the genus Chlamydomonas. All foraminiferal symbionts form a monophyletic group closely related to Chlamydomonas noctigama. The group is composed of seven types identified in this study, including one previously morphologically described species, Chlamydomonas hedleyi. Each of these types can be considered as a separate species, based on the comparison of genetic differences observed between other established Chlamydomonas species. Several foraminiferal species share the same symbiont type, but only one species, Archaias angulatus, was found to bear more than one type. PMID- 11411846 TI - Molecular identification of algal endosymbionts in large miliolid Foraminifera: 2. Dinofiagellates. AB - Large miliolid foraminifers of the subfamily Soritinae bear symbiotic dinoflagellates morphologically similar to the species of the "Symbiodinium" complex, commonly found in corals and other marine invertebrates. Soritid foraminifers are abundant in coral reefs and it has been proposed that they share their symbionts with other dinoflagellate-bearing reef dwellers. In order to test this hypothesis, we have analysed partial large subunit ribosomal DNA sequences from dinoflagellates symbionts obtained from 28 foraminiferal specimens, and compared them to the corresponding sequences of Symbiodinium-like endosymbionts from various groups of invertebrates. Phylogenetic analysis of our data shows that all soritid symbionts belong to the "Symbiodinium" species complex, within which they form seven different molecular types (Frl-Fr7). Only one of these types (Fr1) branches within a group of invertebrate symbionts, previously described as type C. The remaining six types form sister groups to coral symbionts previously designed as types B, C, and D. Our data indicate a high genetic diversity and specificity of Symbiodinium-like symbionts in soritids. Except for type C, we have found no evidence for the transmission of symbionts between foraminifers and other symbiont-bearing invertebrates from the same localities. However, exchanges must have occurred frequently between the different species of Soritinae, as suggested by the lack of host specificity and some biogeographical patterns observed in symbiont distribution. Our data suggest that members of the subfamily Soritinae acquired their symbionts at least three times during their history, each acquisition being followed by a rapid diversification and independent radiation of symbionts within the foraminiferal hosts. PMID- 11411847 TI - Polyamine synthesis and interconversion by the Microsporidian Encephalitozoon cuniculi. AB - Polyamines are small cationic molecules necessary for growth and differentiation in all cells. Although mammalian cells have been studied extensively, particularly as targets of polyamine antagonists, i.e. antitumor agents, polyamine metabolism has also been studied as a potential drug target in microorganisms. Since little is known concerning polyamine metabolism in the microsporidia, we investigated it in Encephalitozoon cuniculi, a microspordian associated with disseminated infections in humans. Organisms were grown in RK-13 cells and harvested using Percoll gradients. Electron microscopy indicated that the fractions banding at 1.051-1.059/g/ml in a microgradient procedure, and 1.102 1.119/g/ml in a scaled-up procedure were nearly homogenous, consisting of pre emergent (immature) spores which showed large arrays of ribosomes near polar filament coils. Intact purified pre-emergent spores incubated with [1H] ornithine and methionine synthesized putrescine, spermidine, and spermine, while [14C]spermine was converted to spermidine and putrescine. Polyamine production from ornithine was inhibitable by DL-alpha-difluoromethylornithine (DFMO) but not by DL-alpha-difluoromethylarginine (DFMA). Cell-free extracts from mature spores released into the growth media had ornithine decarboxylase (ODC), S adenosylmethionine decarboxylase (AdoMetdc), and spermidine/spermine N1 acetyltransferase (SSAT) activities. ODC activity was inhibited by DFMO, but not by DFMA. AdoMetdc was putrescine-stimulated and inhibited by methylglyoxal bis(guanylhydrazone); arginine decarboxylase activity could not be detected. It is apparent from these studies that Encephalitozoon cuniculi pre-emergent spores have a eukaryotic-type polyamine biosynthetic pathway and can interconvert exogenous polyamines. Pre-emergent spores were metabolically active with respect to polyamine synthesis and interconversion, while intact mature spores harvested from culture supernatants had little metabolic activity. PMID- 11411848 TI - Searching for excystment-regulated genes in Sterkiella histriomuscorum (Ciliophora, Oxytrichidae): a mRNA differential display analysis of gene expression in excysting cells. AB - In the absence of food, the oxytrichid Sterkiella histriomuscorum transforms like many ciliates into resting cysts. When transferred back into feeding medium, the cyst re-transforms into a vegetative cell. The entry into and exit from the dormant cyst stage are complex developmental processes still poorly investigated at the molecular level. Assuming that these changes in state could involve changes in gene expression, we have used the technique of mRNA differential display to detect differentially expressed genes in cysts and two different stages of excysting cell. Variation in the temporal expression pattern of transcripts could be detected and, in using an inverse-PCR strategy on circularized macronuclear DNA, we have sequenced the macronuclear genes of three of the isolated cDNAs. which correspond to 1) a nucleotide-binding domain encoding gene, 2) a DHHC-domain-carrying gene, and 3) a phosphatase type 2C encoding gene. For the first two genes, Northern blot analyses supported an excystment-associated regulated gene expression. We discuss their possible role during excystment and we show that the combination of differential display and inverse PCR constitutes a powerful approach to isolate excystment-regulated genes in hypotrichs. PMID- 11411849 TI - Pesticide storage and release in unsaturated soil in Illinois, USA. AB - The chemical fate and movement of pesticides may be subject to transient storage in unsaturated soils during periods of light rainfall, and subsequent release into shallow groundwater by increased rainfall. The objective of this study was to conduct field-scale experiments to determine the relative importance of transient storage and subsequent release of agrichemicals from the vadose zone into potential aquifers. Two field-scale experiments were conducted under a rain exclusion shelter. In the 1x experiment, atrazine and chlorpyrifos were applied at application-rate equivalents (1.6 kg ha(-1) and 1.3 kg ha(-1), respectively). In the 4x experiment, atrazine was applied in an amount that was four times greater than that usually applied to fields (6.7 kg ha(-1)). Water was either applied to simulate rain or withheld to simulate dry periods. In the 1x experiment, atrazine was detected in the water samples whereas chlorpyrifos was not detected in the majority of the samples. The dry period imposed on the treatment plot did not appear to result in storage of the chemicals, whereas the wet period resulted in greater leaching of atrazine, although the concentrations remained less than the Maximum Contaminant Level of 3 microg L(-1). Both chemicals were detected in soil samples collected from a 20- to 30-cm depth, but it appeared that both chemicals dissipated before the field experiment was concluded. It appeared that the one-time application of atrazine and chlorpyrifos at the label rates did not result in a sufficient mass to be stored and flushed in significant concentrations to the saturated zone. When atrazine was applied at 4x and a longer drought period was imposed on the treatment plot, the resulting concentrations of dissolved atrazine were still less than 3 microg L(-1) . Atrazine was detected in only the near-surface (0 to 15 cm) soil samples and the herbicide dissipated before the onset of the dry period in the treatment plot. The results of this field study demonstrated that atrazine and chlorpyrifos were not sufficiently persistent to be stored and then released in significantly large concentrations to the saturated zone. The dissipation half-life of atrazine in the 4x application was about 44 days. This study, in addition to others, suggested that atrazine may be less persistent in surface soil than has been generally reported. PMID- 11411850 TI - Pyrethrins and piperonyl butoxide residues on potato leaves and in soil under field conditions. AB - Residues of pyrethrin-I (Py-I) and pyrethrin-II (Py-II), the major insecticidal components of the pyrethrum daisy (Tanacetum cinerariifolium) as well as residues of piperonyl butoxide (PBO, a pyrethrum synergist) were determined in soil and on potato foliage grown under field conditions. A pyrethrum formulation (Multi Purpose Insecticide) containing the three active ingredients was sprayed twice at the rate of 6 lbs of formulated product.acre(-1) ( 5.4 and 27.2 g A.I. of pyrethrin and PBO, respectively) on potato foliage during the growing season. In soil, three management practices (yard waste compost, grass filter strips, and a no mulch treatment) were used to study the impact of surface soil characteristics on the amount of pyrethrins (Pys) and PBO retained in soil. Soil samples and potato leaves were collected at different time intervals after spraying. Samples were purified and concentrated using solid-phase extraction columns containing C18-Octadecyl bonded silica. Residues were quantified by high-performance liquid chromatograph equipped with a UV detector. The first spray resulted in mean initial deposits of 0.18, 0.40, and 0.99 microg.g(-1) potato leaves for Py-I, Py II, and PBO, respectively. Residues in soil were higher in compost treatments compared to no mulch treatments. PMID- 11411851 TI - Anaerobic biodegradation of DDT residues (DDT, DDD, and DDE) in estuarine sediment. AB - The potential for anaerobic biodegradation of 1,1,1-trichloro-2,2 bischlorophenylethane (DDT), 1,1-dichloro-2,2,-bischlorophenylethane (DDD), and dichlorodiphenylchloroethylene (DDE) in anoxic sediment slurries collected from the Keelung River was investigated in this study. o,p'- and p,p'-DDT were dechlorinated to o,p'- and p,p'-DDD, respectively, and then transformed to other compound(s). 1-Chloro-2,2-bis (p-chlorophenyl) ethylene (DDMU) and trace amount of dichlorobenzophenone (DBP) were detected in sediment slurries amended with p,p'-DDT or p,p'-DDD. DDMU was also detected in sediment slurries amended with p,p'-DDE. The relative transformation rates for both o,p'- and p,p'-isomers of DDT, DDD, and DDE were DDT>DDD>DDE. Re-addition of DDT, DDD, or DDE to the sediment slurries after initial removal enhanced the respective dechlorination rates. The transformation rates of the p,p'-isomers of both DDT and DDD were faster than those of the respective o,p'-isomers. p,p'-DDT dechlorination in the p,p'-DDT-adapted sediment slurries were inhibited by the addition of molybdate, or molybdate plus sulfate, but not inhibited by the addition of sulfate. Addition of bromoethane-sulfonic acid (BESA) slightly inhibited p,p'-DDT dechlorination. Non-adapted sediment slurries lost the ability to dechlorinate pentachlorophenol during adaptation to p,p'-DDT. p,p'-DDD was the major transformation product of p, p'-DDT in 3,4,4',5-tetrachlorobiphenyl-adapted sediment slurries, which suggested that the microbial community in the 3,4,4',5-CB-adapted sediment was unable to remove chlorine from the aromatic rings of p,p'-DDT. PMID- 11411852 TI - Metabolism of fluvalinate in chickpea plants under sub-tropical conditions of northern India. AB - The metabolism of fluvalinate in chickpea plants in the field under sub-tropical climatic conditions has been investigated. Ten days after spraying with fluvalinate, leaves were collected and extracted in acetone. The extract was fractionated and the metabolites were identified by co-chromatography using TLC, GLC, and GC-MS. The anilino acid, a metabolite derived from ester cleavage, was the major metabolite present as both free and conjugated forms in chickpea leaves. 3-PBacid and 3-PBalc were minor metabolites. Glucose was found to be the major sugar conjugating with the metabolites. PMID- 11411853 TI - Anaerobic treatment of atrazine bearing wastewater. AB - Performance of mixed microbial anaerobic culture in treating synthetic waste water with high Chemical Oxygen Demand (COD) and varying atrazine concentration was studied. Performance of hybrid reactors with wood charcoal as adsorbent, with a dose of 10 g/l and 40 g/l, along with the microbial mass was also studied. All the reactors were operated in sequential mode with Hydraulic Retention Time (HRT) of 5 days. In all the cases, COD removal after 5 days was found to be above 81%. Initial COD was above 1,000 mg/l. From a hybrid reactor COD removal after 2 days was observed to be 90%. Atrazine reduction after 5 days by microbial mass alone was 43.8%, 40% and 33.2% with an initial concentration of 0.5, 1.0 and 2.0 mg/l respectively. MLSS on all the cases were almost same. Increasing MLSS concentration by about 2 fold did not increase the atrazine removal efficiency significantly. Maximum atrazine removal was observed to be 64% from the hybrid reactor with 10 g/l of wood charcoal and 69.4% from the reactor with 40 g/l of wood charcoal. Atrazine removal from the hybrid reactors after 15 days were observed to be 35.7% and 38.7%, which showed that the higher dose of wood charcoal in hybrid reactor did not improve the atrazine removal efficiency significantly. Specific methanogenic activity test showed no inhibitory effect of atrazine on methane producing bacteria. The performance of anaerobic microorganisms in removing atrazine with no external carbon source and inorganic nitrogen source was studied in batch mode. With an initial concentration of 1.0 mg/l, reduction of atrazine by the anaerobic microorganisms in absence of external carbon source after 35 days was observed to be 61.8% where as in absence of external carbon and inorganic nitrogen source the reduction was only 44.2% after 150 days. Volatilization loss of atrazine was observed to be insignificant. PMID- 11411854 TI - Homogenous catalyzed ozonation of simazine. Effect of Mn(II) and Fe(II). AB - Simazine, [2-chloro, 4,6-bis(ethylamino)-1,3,5-s-triazine], a common herbicide typically found in surface and ground water was ozonised by using catalytic amounts of Mn(II) and Fe(II). An optimum value for metal concentration was found in the process. Some inhibition of the simazine degradation rate was observed when these metals were added above the optimum concentration. The pH of the reaction media played a significant role in the simazine oxidation rate. Thus, an increase in simazine conversion was observed when the pH was raised from 5 to 9. However, the catalytic effect of added manganese was negligible at the highest pH used in this study (pH 9). The beneficial influence of Fe(II) was also observed when utilising the combination of ozone and hydrogen peroxide. Contrarily, Mn(II) presented a negative influence on simazine conversion when using this oxidation technology. PMID- 11411855 TI - Comparison of in vitro activities of amphotericin, clotrimazole, econazole, miconazole, and nystatin against Fusarium oxysporum. AB - The inhibitory activity of amphotericin B, clotrimazole, econazole, miconazole and nystatin was compared against Fusarium oxysporum f.sp. radicis-cucumerinum. The most efficient antifungal agent against the growth of Fusarium oxysporum was econazole, followed by clotrimazole, miconazole, amphotericin and nystatin. The ED50 and ED90 values were 0.053 and 1.002 ppm for econazole, 0.088 and 1.100 ppm for clotrimazole, 0.173 and 3.210 ppm for miconazole, 0.713 and greater than 48 ppm for amphotericin and 3.860 and 16.702 ppm for nystatin. The ED50 values of nystatin and amphotericin against spore germination of Fusarium oxysporum were determined at 3.1427 ppm and 8.3990 ppm respectively, nystatin was 2.76 times more effective than amphotericin, while no effect was observed after the addition of econazole, clotrimazole and miconazole. The tested azoles were more effective than amphotericin and nystatin on growth inhibition of Fusarium oxysporum but amphotericin and nystatin acted significantly better on spore germination of Fusarium. PMID- 11411856 TI - Treatment of greenhouse wastewater using constructed wetlands. AB - Five wetland designs, based on conventional surface flow (SF) and subsurface flow (SSF) approaches, were assessed for nitrogen and phosphorus removal from greenhouse wastewater. Results indicated none of the individual designs assessed was capable of providing the highest treatment effect for all nutrients of concern; however, the SF wetland emerged as the most appropriate design for the treatment of greenhouse wastewater. The highest mean phosphorus reduction of 65% was observed in the unplanted SF wetlands. Peak nitrate reductions of 54% were observed in the 15-cm deep SF wetlands and ammonia removal of 74% was achieved in the unplanted SF wetlands. Nitrate concentration in the greenhouse effluent can be reduced to acceptable levels for the protection of freshwater aquatic life (i.e., less then 40 ppm) using a loading rate of 1.65 g NO3-N/m2/day and a design water depth of 30 cm or greater. Based on available literature and the results of this research project, a multistage design, consisting of an unplanted pre treatment basin followed by a 25 to 35 cm deep surface flow marsh with open water components, is recommended. PMID- 11411857 TI - Methodology for management of endosulfan contaminated eluent. AB - Management of Endosulfan contaminated eluent (24 mg/l) resulting from a treatment process to remove Endosulfan from water with wood charcoal, was attempted using various methods viz. volatilisation, hydrolysis and sorption by viable cell bacteria with and without acclimatisation. Volatilisation failed in giving better result, as Endosulfan was not considerably volatile. It could achieve a removal efficiency of 1.4-2%. Hydrolysis resulted in 28.4% and 17.9% removal of Endosulfan in acidic and alkaline media, respectively. Viable cell bacteria (aerobic) without prior acclimatization showed efficiency of 89.7% and after prior acclimatisation showed 96% removal efficiency. Sorption by the acclimatized biomass was found a suitable method for the removal of Endosulfan at a concentration of 24 mg/l. PMID- 11411858 TI - Characterization of activated carbons prepared from sugarcane bagasse by ZnCl2 activation. AB - Activated carbons were prepared from the agricultural waste of sugarcane bagasse by the chemical activation with zinc chloride (ZnCl2) at the activation temperature of 500 degrees C with soaking time of 0.5 hour. The influence of activation parameters on the final carbon products was examined by varying the impregnation ratio (i.e., mass ratio of added ZnCl2 to bagasse) and bagasse size. The physical properties of carbon products were characterized by nitrogen adsorption/desorption isotherms (at 77 K) and helium displacement method. The surface area and pore volume of carbons were thus obtained by the BET equation and t-plot method. Also, the particle density and porosity of carbons were estimated by the total pore volume and true density. The increases of the values of surface area and pore volume are approximately proportional to the impregnation ratio. The microporous carbon product with the BET surface area of 905 m2/g and total pore volume of 0.44 cm3/g was obtained in the present study. Further, the adsorption isotherms of two acid dyes from aqueous solutions onto the carbon products were performed at 30 degrees C. The results show that the adsorption isotherms of acid dyes with high molecular weight or large molecular size on the microporous adsorbents of activated carbons are plateau forms, indicating multilayer adsorptions, which may be attributed to the steric hindrance of the adsorbate molecules. PMID- 11411859 TI - Fluorescent RT in situ PCR detection of MRP1 mRNA in human HCV infected liver. PMID- 11411860 TI - The 2001 Maffo Vialli International Award for Histochemistry. PMID- 11411861 TI - Nuclear changes in pathogen-infected tomato roots. AB - We have investigated nuclear changes induced in tomato (Lycopersicon esculentum) plant roots by two different pathogens, a phytoplasma of the stolbur group and the soil-borne fungus Phytophthora nicotianae var. parasitica, using light and electron microscopy as well as flow cytometry. Our results show that both pathogens strongly influenced nuclear structure, causing pyknosis and chromatolysis, and induced variations in proportion of nuclear populations with different DNA content, increasing the 2C and decreasing the 4C nuclear populations. These latter results suggest a block of the endoreduplication process in the presynthetic phase of the host cell cycle. Moreover, in pathogen infected roots, nuclei of the same DNA content were less fluorescent, and therefore had a lower DNA content, compared to those of controls, as confirmed by morphological analyses. Our findings suggest that different stimuli may evoke similar nuclear changes, and confirm the usefulness of flow cytometry as a tool to recognize the stress conditions related to the presence of a pathogen. PMID- 11411862 TI - Spatial expression of DNA topoisomerase I genes during cell proliferation in Daucus carota. AB - The spatial expression of carrot (Daucus carota L.) top1 genes encoding the two isoforms of the enzyme DNA topoisomerase I (EC 5.99.1.2) was investigated. In situ hybridization analysis performed with a probe recognizing both top1 transcripts provided evidence that in explanted hypocotyls induced to proliferate in vitro by the addition of the growth regulator 2,4-dichlorophenoxyacetic acid (2,4-D), the mRNA accumulation parallels the proliferation of provascular cells of the stelar cylinder. During somatic embryogenesis, the histological distribution of top1 transcripts was strongly evident at the stage of torpedo shaped embryos, but gene expression was not only restricted to meristematic regions. When the spatial localization was extended to carrot vegetative apices and the investigation was carried out with specific probes for top1alpha and top1beta, both transcripts preferentially accumulated in tissues having mitotic activity. PMID- 11411863 TI - Extracellular polysaccharides are involved in the attachment of Azospirillum brasilense and Rhizobium leguminosarum to arbuscular mycorrhizal structures. AB - Arbuscular mycorrhizal (AM) fungi, one of the most important component of the soil microbial community, establish physical interactions with naturally occurring and genetically modified bacterial biofertilizers and biopesticides, commonly referred to as plant growth-promoting rhizobacteria (PGPR). We have used a genetic approach to investigate the bacterial components possibly involved in the attachment of two PGPR (Azospirillum and Rhizobium) to AM roots and AM fungal structures. Mutants affected in extracellular polysaccharides (EPS) have been tested in in vitro adhesion assays and shown to be strongly impaired in the attachment to both types of surfaces as well as to quartz fibers. Anchoring of rhizobacteria to AM fungal structures may have special ecological and biotechnological significance because it may facilitate colonisation of new rhizospheres by the bacteria, and may be an essential trait for the development of mixed inocula. PMID- 11411864 TI - The year of encroaching genomics. PMID- 11411865 TI - Cellulose in algal cell wall: an "in situ" localization. AB - In order to ascertain the presence and the in muro localization of cellulose, the enzyme-gold affinity test was applied to algal cell walls. The high specificity of affinity cytochemistry allowed us, by using the enzyme cellulase, to confirm the available biochemical data and to give a map of the cellulose localization in different algal groups. Taking into account the complex skeletal polysaccharide structure and composition of the algal cell walls, this method proved to be a reliable tool in this field. PMID- 11411866 TI - Dipeptidyl peptidase IV in two human glioma cell lines. AB - There is growing evidence that dipeptidyl peptidase IV [DPP-IV, EC 3.4.14.5] takes part in the metabolism of biologically active peptides participating in the regulation of growth and transformation of glial cells. However, the knowledge on the DPP-IV expression in human glial and glioma cells is still very limited. In this study, using histochemical and biochemical techniques, the DPP-IV activity was demonstrated in two commercially available human glioma cell lines of different transformation degree, as represented by U373 astrocytoma (Grade III) and U87 glioblastoma multiforme (Grade IV) lines. Higher total activity of the enzyme, as well as its preferential localisation in the plasma membrane, was observed in U87 cells. Compared to U373 population, U87 cells were morphologically more pleiomorphic, they were cycling at lower rate and expressing less Glial Fibrillary Acidic Protein. The data revealed positive correlation between the degree of transformation of cells and activity of DPP-IV. Great difference in expression of this enzyme, together with the phenotypic differences of cells, makes these lines a suitable standard model for further studies of function of this enzyme in human glioma cells. PMID- 11411867 TI - DNA aberrations in urinary bladder cancer detected by flow cytometry and FISH: prognostic implications. AB - We evaluated the genetic changes in bladder cancer biopsy by fluorescence in situ hybridization (FISH) and related them to stage and grade of the tumor, ploidy (FCM) and clinical outcome, to determine a simple method to identify tumors with a poorer prognosis. Using FISH the numerical aberrations of chromosomes 1, 7, 9, 17 in tumor's imprints of 70 patients with transitional cell cancer (TCC) were determined. First of all, the data demonstrated that the sensitivity of FISH in detecting quantitative DNA aberrations exceeds FCM's sensitivity. The frequency of chromosome 1 and 9 aberrations did not show significant differences in diploid and aneuploid tumors in different stage and grade. On the contrary, the chromosome 7 and 17 aneusomy showed greater differences between pT1 and pT2-3 tumors (p<0.032 and p<0.0006, respectively) than between stage pTa and pT1. In our investigation, an increasing number of aberrations was observed in all chromosomes examined in tumors of patients who afterwards underwent cystectomy and/or had recurrent tumors. These results suggest that chromosome 7 and 17 aneusomy could be predictive of adverse outcome in a subgroup of patients with superficial tumors at presentation. PMID- 11411868 TI - Prognostic factors affecting disease-free survival rate following surgical resection of primary breast cancer. AB - In order to identify the prognostic factors that significantly influence the disease-free survival rate after surgical resection of primary breast cancers, we determined tumour and lymph node grades, and immunohistochemical staining for estrogen and progesterone receptors (ER and PR), c-erbB-2, p53, bcl-2, bax and PCNA in 76 patients. Univariate analysis showed that increased grade of tumour and lymph nodes, negative immunostaining for ER, positive immunostaining for c erbB-2, and a high PCNA index (> or = 30%) negatively influenced the disease-free survival rate, but PR, p53, bcl-2 and bax had no predictive value. Although p53 was not an independent prognostic factor by itself, the combination of p53, bcl 2, and bax proved to correlate with the disease-free survival, with the best prognosis noted in tumours negative for p53 and positive for both bcl-2 and bax, intermediate prognosis in tumours negative for p53 and positive for either bcl-2 or bax and worst prognosis in tumors negative for p53 as well as bcl-2 and bax. Tumour grade correlated positively with PCNA index, while positive staining for ER correlated negatively with tumour grade as well as with PCNA index, although this was statistically insignificant. Immunostaining of breast cancers for bcl-2 correlated negatively with tumour grade and PCNA index. Immunostaining for c-erbB 2 correlated positively with PCNA but not with tumour grade. Immunostaining for p53 tended to correlate positively with PCNA, but not with tumour grade. Immunostaining for PR and bax did not correlate with tumour grade and PCNA index. These results suggest that in addition to tumour size and lymph node involvement, immunostaining for ER, c-erbB-2, and a high PCNA index are important prognostic factors in human breast cancer. Wild-type p53 with preserved bcl-2 and bax gene products is also a favorable prognostic factor indicating breast cancer at an early stage of cancer progression. PMID- 11411869 TI - Lymphocyte dysmetabolism: an immunocytochemical comparative approach in IDDM and control subjects. AB - We have investigated by immuno-electron microscopy the presence of phosphotyrosine in cells as a whole and in different cell districts (nucleus, cytoplasm, plasma membrane, and mitochondria) in peripheral blood lymphocytes of IDDM (insulin-dependent diabetes mellitus) patients and age-matched controls. Immuno-gold particle density was highest in mitochondria and decreased in cytoplasm, nucleus and plasma membrane. The time dependence of phosphotyrosine labelling after cell isolation was very strong in all subcellular populations, with a fall in immunogold staining after 30 min. Staining levels at zero time were similar in controls and IDDM patients; the loss of phosphotyrosine labelling was much stronger in controls, except in the plasma membrane. Plasma membrane NADH oxidoreductase activity, studied using cytosolic NADH as substrate and assayed with DCIP as acceptor, was significantly increased in IDDM patients, suggesting a response to a deficient mitochondrial energetic activity. The fact that NADH oxidoreductase is a growth factor related to tyrosine phosphorylation pathways raises intriguing questions on the cellular derangement occurring in peripheral lymphocytes in IDDM, although the relationships among the immunocytochemical and biochemical changes is still obscure. PMID- 11411870 TI - The nucleus of differentiated root plant cells: modifications induced by arbuscular mycorrhizal fungi. AB - The nuclei of plant cells show marked differences in chromatin organisation, related to their DNA content, which ranges from the type with large strands of condensed chromatin (reticulate or chromonematic nuclei) to one with mostly decondensed chromatin (chromocentric or diffuse nuclei). A loosening of the chromatin structure generally occurs in actively metabolising cells, such as differentiating and secretory cells, in relation to their high transcriptional activity. Endoreduplication may occur, especially in plants with a small genome, which increases the availability of nuclear templates, the synthesis of DNA, and probably regulates gene expression. Here we describe structural and quantitative changes of the chromatin and their relationship with transcription that occur in differentiated cells following an increase of their metabolism. The nuclei of root cortical cells of three plants with different 2C DNA content (Allium porrum, Pisum sativum and Lycopersicon esculentum) and their modifications induced by arbuscular mycorrhization, which strongly increase the metabolic activity of colonised cells, are taken as examples. PMID- 11411871 TI - Three-dimensional computer-aided reconstruction of FMRFamide immunopositive neuron distribution in the ventral ganglion of the barnacle Balanus amphitrite (Cirripedia, Crustacea). AB - We have implemented a simple program to solve three of the problems related to 3D reconstruction (3D-Rec) of soft tissues: alignment of sections, distortions, and estimation of the spatial position of elements of interest inside the tissues. As a model, we chose the distribution of FMRFamide-like immunopositive neurons in the ventral ganglion of the barnacle Balanus amphitrite collected during different seasonal periods. Images of immunostained sections were acquired by means of a CCD-camera-equipped microscope and a PC and the reference points were taken inside the sections. The FMRFamide-like immunopositive neurons detected in the barnacle ventral ganglion were grouped into four different classes according to size, shape and staining intensity. More numerous FMRFamide-like immunopositive neurons were detected in the autumn-collected barnacle than in the summer counterpart. The two 3D reconstructions obtained from transverse and longitudinal ventral ganglion sections were efficaciously compared after 90 degrees rotation of one of them. Comparison of these two 3D-Rec suggests the presence of at least two groups of FMRFamide-like immunopositive neurons that are seasonally-related and probably involved in reproduction. PMID- 11411872 TI - Hip pain in the young adult: diagnosis and treatment of disorders of the acetabular labrum and acetabular dysplasia. AB - Hip pain in the young adult patient (ages, 17-35 years) is a diagnostic and therapeutic challenge for the orthopedic surgeon. The diagnostic and treatment algorithm for many causes of hip pain in the young adult-such as tumors and tumoral conditions, rheumatoid and degenerative arthritis, osseous necrosis, and the sequelae of congenital problems such as slipped capital femoral epiphysis and Legg-Calve Perthes--are well described and established. However, the diagnosis and treatment algorithm for disease processes, such as disorders of the acetabular labrum and dysplasia of the hip, are less clear. Advances in diagnostic radiology, particularly magnetic resonance arthrography, and advances in therapy, including the successful use of the Bernese periacetabular osteotomy and hip arthroscopy, have allowed for both timely diagnosis and appropriate treatment of these sources of hip pain in the young adult. The purpose of this review is to outline the diagnostic and treatment decision-making protocol for young adult patients with hip pain, and specifically to discuss the treatment of acetabular labral tears and dysplasia of the hip. PMID- 11411873 TI - Temporal stability of the Minnesota Multiphasic Personality Inventory (MMPI) in patients undergoing lumbar fusion: a poor predictor of surgical outcome. AB - In this prospective study, the Minnesota Multiphasic Personality Inventory (MMPI) was administered to patients before and after lumbar spines fusion to investigate the stability of MMPI scores after surgical intervention and to attempt to correlate MMPI scale scores with outcome data. Sixty-eight patients were included. Testing was performed before surgery and at a mean of 1.5 years after surgery. Clinical outcome ratings were assigned by using criteria of pain relief and analgesic use. In addition, demographic variables known to affect outcome were analyzed. Sixty percent of the patients had a successful clinical outcome. Positive outcome correlated with the demographic factors of occupation (homemaker) and solid fusion. MMPI scales were stable across time, with no difference between groups. Independent t tests were used to study preoperative MMPI scores with respect to clinical outcome. Unsatisfactory outcomes were associated with higher scores on scales 1, 3, and 10 before surgery. Postoperative testing revealed significant outcome correlations--higher scores on scales 1, 2, 3, 5, 7, and 8 associated with an unsatisfactory outcome. However, discriminant function analysis of preoperative MMPI data was able to classify outcomes correctly in only 58.8% of the cases. The utility of the MMPI as a predictor of outcome after surgical intervention is quite limited. Use of group data and testing before and after surgery does not appear to influence this conclusion. Although the scores as a group were stable across time, the amount of variance in outcome that could not be accounted for by using MMPI scales as predictors was unacceptably large. PMID- 11411874 TI - Complications in limb-lengthening procedures: a review of 49 cases. AB - The purpose of this study was to examine the nature and number of complications relative to magnitude of limb lengthening. Results and complications of 50 limb lengthening procedures were analyzed. There were 49 patients. Mean age was 21 years (range, 7-48 years). Lengthening was performed in 24 femora, 18 tibiae, 4 humerii, 3 radii, and 1 ulna. Average length gained was 5 cm (range, 3-15 cm) at average follow-up of 48 months (range, 12-76 months). Desired length was accomplished in all but 1 patient. The 69 complications varied in severity, but only 5 of these significantly impaired end results. The most serious complications occurred in patients with >30% bone lengthening. Patients with <15% lengthening had a significantly decreased complication rate. The healing index was lower in children. We conclude that incidence and severity of complications after limb-lengthening procedures are significantly influenced by relative lengthening of bone. PMID- 11411875 TI - Smoking and bony union after ulna-shortening osteotomy. AB - We examined the relation between cigarette use and occurrence of delayed union and nonunion after ulna-shortening osteotomy for ulnar impaction syndrome. After obtaining the smoking history of 39 patients (40 wrists), we found that, compared with nonsmokers, smokers experienced longer healing times and more of them experienced healing delay and nonunion. Mean union rates were 7.1 months in smokers and 4.1 months in nonsmokers. Six smokers (30%) and no nonsmokers experienced delayed union or nonunion. Given the adverse effects of smoking on bony union, we recommend that smoking history be considered when selecting patients for ulna-shortening procedures. PMID- 11411876 TI - Madura foot: treatment of Nocardia nova infection with antibiotics alone. AB - This case report documents the presentation of a 29-year-old woman with a deep Nocardia nova infection of the foot involving the bones. The patient responded well to prolonged antibiotic therapy, with essentially complete resolution of her symptoms. The results suggest that surgical intervention in these unusual cases may be unnecessary and that good clinical results can be obtained pharmacologically. PMID- 11411877 TI - Bilateral metachronous rupture of the patellar tendon. AB - We present a case of a 37-year-old man who sustained a rupture of his left patellar tendon approximately 48 hours after rupturing his right patellar tendon. This temporal pattern illustrates two important aspects of patellar tendon injury that rupture of the degenerated patellar tendon may occur without any prodromal warning and that it may elude detection even if the patient is examined by several physicians. PMID- 11411878 TI - Complete dorsal tarsometatarsal dislocation of the first metatarsal. AB - The authors describe a rare case of Lisfranc's dislocation that occurred after a 13-year-old boy kicked the ground while playing soccer. He presented with complete pure dorsal tarsometatarsal dislocation of the first metatarsal. He was treated succesfully by closed reduction and percutaneous K-wire fixation. No complication was found and he returned to his preinjury lifestyle at 3 months. No sequelae are found 3.5 years later. PMID- 11411879 TI - Madelung's deformity. PMID- 11411880 TI - York review. PMID- 11411881 TI - Sporty but safe. PMID- 11411882 TI - Microbial aerosols. PMID- 11411883 TI - Blowing the whistle. PMID- 11411884 TI - Privatised dental care. PMID- 11411885 TI - Training needs analysis. PMID- 11411886 TI - Mortality associated with odontogenic infection! AB - Odontogenic causes are the most common source for spreading maxillo-facial infections. These infections can develop into life threatening events. However a fatal outcome is fortunately rare and is generally associated with an immunocompromised status. This case report highlights a spreading maxillo-facial infection, which resulted in massive haemorrhage from the subclavian vein into the pleural cavity and subsequent death of a young fit male patient. PMID- 11411887 TI - Current principles in the management of obstructive sleep apnoea with mandibular advancement appliances. AB - This paper looks at the role of mandibular advancement splints in the management of obstructive sleep apnoea, as part of a multidisciplinary approach. Clinical guidance is provided on the rationale for use, patient selection, design features and the associated risk to benefit ratio. PMID- 11411888 TI - A three-part bilateral swinglock design denture revisited. AB - This case was first reported by one of the authors in 1987, subsequent to the patient being treated using a combination of a hinge and split pin sectional denture. The same patient attended for review some 14 years later for assessment of this denture at which stage a replacement was made. PMID- 11411889 TI - Inequalities in availability of National Health Service general dental practitioners in England and Wales. AB - AIM: To model the inequalities in availability of National Health Service general dental practitioners in England and Wales in relation to key socio-demographic factors. METHODS: Current estimates of the numbers of NHS general dental practitioners for each health authority were related to data from the 1991 census using Poisson regression models, and generalised estimating equations to allow for correlation between results for neighbouring health authorities. RESULTS: An 'average' health authority, without a dental school, would be expected to have 2,138 residents for every NHS dentist. Controlling for relevant factors, health authorities with higher proportions of the following are associated with lower (better) population to dentist ratios by the amounts shown: each 1% higher female population (-11.8%; 95%CI -19.1%, -3.9% P = 0.004); each 1% greater South Asian population (-1.4%; 95%CI -2.1%, -0.7% P <0.001). A health authority with a dental school is associated with a more favourable ratio compared with one without such a facility (-9.2%; 95%CI -16.2%, -1.6% P = 0.019). Each additional 1% of the following are associated with a worse ratio by the amounts shown: children aged 0 to 14 years old (+5.2%; 95% CI +2.4%, +8.1% P < 0.001); adults aged over 65 years old (+2.8%, 95%CI +1.0%, +4.7% P =0.002); households without a car (+0.8%; 95%CI 0.0%, +1.6% P =0.042). CONCLUSIONS: Ensuring access to dental care may be a more complex issue than simply providing adequate numbers of dentists at a national level. Any manpower planning exercise should additionally consider local factors that may act as incentives or disincentives to those professionals who provide care. PMID- 11411890 TI - Evaluation of computer aided learning in developing clinical decision-making skills. AB - AIM: The aim of this study was to determine whether an educational intervention delivered by a computer aided learning package improved the sensitivity and specificity of dentists' restorative treatment decisions. METHOD: The study was a randomised controlled study using a Solomon three-group design. Ninety-five dentists were randomly allocated to the three study groups. One group of dentists read the radiographs pre and post an educational intervention, a second group read the radiographs once, after the intervention, and a third group read the radiographs twice, but received no intervention. On each occasion the dentists read 24 surfaces on each of 15 radiographs and made 360 decisions on how certain they were about restoring the tooth surface. Comparisons of mean sensitivity, specificity and areas under ROC curves were made within and between the study groups. Kappa values were used to assess changes in the level of agreement between dentists. RESULTS: There were no significant changes in sensitivity, specificity or area under ROC curves caused by the intervention. There was no evidence that the level of agreement between the dentists improved after the intervention. CONCLUSION: A computer aided learning package had no effect on dentists' treatment decision-making behaviour. PMID- 11411891 TI - Mercury vapour release from a dental aspirator. AB - OBJECTIVE: To investigate the release of mercury vapour from a dental aspirator which vented its waste air through its base directly into the surgery environment. METHODOLOGY: Mercury vapour in air concentrations were measured at the breathing zone of the dentist during continuous operation of the aspirator. Further series of mercury vapour measurements taken at the aspirator exhaust vent were carried out to determine the sources of mercury vapour from this particular device. RESULTS: At the dentist's breathing zone, mercury vapour concentrations of ten times the current occupational exposure limit of 25 micrograms/m3 were recorded after 20 minutes of continuous aspirator operation. A build up of amalgam contamination within the internal corrugated tubing of the aspirator was found to be the main source of mercury vapour emissions followed by particulate amalgam trapped within the vacuum motor. As the vacuum motor heated up with run time, mercury vapour emissions increased. It was found that the bacterial air exhaust filter (designed to clean the contaminated waste air entering the surgery) offered no protection to mercury vapour. In this case the filter trapped particulate amalgam which contributed to further mercury vapour contamination as high volume air was vented through it. CONCLUSION: It is not known how many dental aspirators are in use that vent their waste air directly into the surgery or if this aspirator is representative of others in existence. The safety of dental aspirating systems with regard to mercury vapour exposure requires further investigation. PMID- 11411892 TI - An evaluation of the flammability of five dental gloves. AB - OBJECTIVE: To assess the flammability of five brands of dental procedure glove. DESIGN: A total of ten gloves of each brand (Biogel-D, Premier Protectors, Roeko D, Safeskin Satin Plus and Schottlander Low Allergy) underwent flammability testing. Five of these were tested as supplied (unwashed) and five following the application of the hand disinfectant Hydrex (washed). Each glove was stretched over a metal frame and the time to ignition, when exposed to a standardised butane flame, recorded. In addition, the thickness of each glove was also assessed by micrometer measurement. RESULTS: All the gloves tested ignited in less than 2.5 seconds. Two way analysis of variance revealed significant effects of glove type (P< 0.001) and treatment (P< 0.05), together with a significant interaction of these factors (P < 0.05), upon the ignition time. Washing Roeko-D gloves with Hydrex significantly (P< 0.01) retarded the ignition time compared with those in the unwashed state. Both the glove thickness and material type appeared to be related to ignition time. CONCLUSION: The work presented here demonstrates the acute and varying flammability of a range of dental procedure gloves. It should serve as a reminder to those who routinely use open flames whilst gloved of the dangers of this practice. PMID- 11411893 TI - Spirits having flown. PMID- 11411894 TI - Formulation and physiological and biopharmaceutical issues in the development of oral lipid-based drug delivery systems. AB - The rapidly increasing availability of drug receptor structural characteristics has permitted the receptor-guided synthesis of potential new drug molecules. This synthesis strategy frequently results in the creation of polycyclic and highly hydrophobic compounds, with attendant poor oral bioavailability resulting from low solubility and slow dissolution rate in the primarily aqueous contents of the gastrointestinal (GI) tract. In an attempt to improve the solubility-limited bioavailabiliy associated with these compounds, formulators have turned to the use of lipid excipients in which the compounds can be solubilized prior to oral administration. This new class of excipients presents the pharmaceutical scientist with a number of new challenges at all stages of the formulation development process, beginning with the excipient selection and stability assessment of the prototype formulation, up to and including scale-up and mass production of the final market-image product. The interaction of lipid-based formulations with the gastrointestinal system and associated digestive processes presents additional challenges and opportunities that will be understood more fully as we begin to unravel the intricacies of the GI processing of lipid excipients. For example, an increasing body of evidence has shown that certain lipids are capable of inhibiting both presystemic drug metabolism and drug efflux by the gut wall mediated by p-glycoprotein (PGP). And, it is well known that lipids are capable of enhancing lymphatic transport of hydrophobic drugs, thereby reducing drug clearance resulting from hepatic first-pass metabolism. This review addresses the current state of knowledge regarding oral lipid-based formulation development and scale-up issues and the physiological and biopharmaceutical aspects pertinent to the development of an orally bioavailable and efficacious dosage form. PMID- 11411895 TI - Methods for the recovery and purification of polyene antifungals. AB - Despite the development of newer antifungal drugs, the polyene antifungals continue to be the most potent broad-spectrum fungicides available for clinical use. The incidence and severity of fungal infections are on the rise, underscoring the need for new and more effective antifungal drugs. Thus, the search for new polyene antifungals is ongoing. The limited solubility, polymorphic character, and inherent chemical instability of these compounds make their economical recovery and purification from mass culture challenging problems in biotechnology. This article provides a comprehensive review of the methods that have been developed for the recovery and purification of amphotericin B and nystatin, the two most important polyenes currently in clinical use. PMID- 11411896 TI - Naproxen particle design using porous starch. AB - Naproxen (Nap) was embedded in porous starch by preferential grinding, and we examined the physicochemical properties of these particles, including pore diameter, pore volume, and dissolution of naproxen. Porous starch (PS) particles made by preferential grinding with a Mechanofusion system had a higher content of naproxen than those made using the Mechanomill as determined using a mercury porosimeter. Neither sample showed any significant changes in crystallization state of naproxen in particles as determined by powder X-ray diffraction and differential scanning calorimetry (DSC). No interactions occurred between naproxen and porous starch due to preferential grinding as determined by powder X ray diffraction and DSC. The dissolution rate of drug from particles prepared by preferential grinding was faster than that from physical mixtures. PMID- 11411897 TI - The development of a blend-sampling technique to assess the uniformity of a powder mixture. AB - This article discusses the challenges overcome during the development of a blend sampling technique and the successful validation of the blending operation for a tablet dosage form containing 2% active ingredient. Content uniformity results are discussedfor three pilot-scale (15-kg) and seven commercial-scale (150-kg) batches of tablets. Blend and core content uniformity data from the pilot-scale batches were acceptable. For the initial commercial-scale batches, although the tablet core content uniformity data were acceptable, the blend uniformity results were poor. The blend data for these batches had very high mean values, but acceptable relative standard deviations (RSDs). This suggested that the drug was being preferentially sampled by the thief but in a consistent, reproducible manner. Extensive testing was performed on a commercial-scale development batch to identify potential causes of sampling error. The results of this testing helped define the blend-sampling technique and strategy used to validate the mixing operation. PMID- 11411898 TI - Preparation and physical characterization of alginate microparticles using air atomization method. AB - Alginate microparticles were prepared using an air atomization method and varying formulation and processing conditions. Thereafter, the size and surface morphology of alginate microparticles were characterized. The trapping efficiencies of the ketoconazole, acetaminophen, vitamin C, and Bifidobacteria bifidum as model core materials were then determined. The air atomization process produced free-flowing and small-size microparticles after the freeze-drying process. The size distribution and surface morphology varied depending on the concentration of wall-forming materials and processing conditions. Generally, the geometric mean size increased as the concentration of alginate and poly-1-lysine and the delivery rate increased, but the air pressure decreased. Most of all, the ratio of delivery rate of alginate solution and air pressure could affect the size and surface morphology of alginate microparticles. However, the geometric mean size of alginate poly-1-lysine microparticles reproducibly ranged from about 80 to 130 microm. The microparticles were irregularly spherical or elliptical. The trapping efficiencies of ketoconazole, acetaminophen, vitamin C, and bifidobacteria were determined to be 71.5%, 60.1%, 1.6%, and 31%, respectively, when alginate concentration (1.5%), poly-1-lysine concentration (0.02%), air pressure (0.75 bar), delivery rate (8 ml/min), and spraying distance (45 cm) were applied. The current microencapsulation process using the air atomization method provides an alternative to entrapping small molecules and macromolecules without using harmful organic solvents. In addition, the small-size and free-flowing alginate microparticles containing active substances can be used as an intermediate in pharmaceutical applications. PMID- 11411899 TI - Preformulation studies on the S-isomer of oxybutynin hydrochloride, an Improved Chemical Entity (ICE). AB - (S)-Oxybutynin HCl (S-OXY) is a white crystalline solid powder with an acicular particle morphology. Differential scanning calorimetry (DSC) thermograms revealed one characteristic endotherm at 116.2 degrees C. On rescanning a sample heated to 120 degrees C, no thermal events were distinguished in the temperature range 25 degrees C to 150 degrees C. Weight loss curves determined by thermogravimetric analysis showed a continuous, gradual weight loss of about 0.15% over the temperature range 30 degrees C to 110 degrees C, followed by a change in slope and more rapid weight loss beginning at 150 degrees C. Observation by hot-stage microscopy confirmed the melting endotherm observed by DSC. Equilibrium moisture uptake studies indicated low water vapor uptake at low relative humidities (<52.8%). At relative humidities of 75.3% and 84.3%, S-OXY first deliquesced and then converted to a lower melting point crystal form. X-ray powder diffraction (XRPD) data supported the DSC findings. S-OXY underwent degradation by ester hydrolysis at alkaline pHs. The kinetics of this reaction were studied at 25 degrees C in carbonate-bicarbonate buffers. Observed rate constants of 0.008 h( 1) and 0.0552 h(-1) were determined at pH 9.69 and 10.25, respectively. The pKa of S-OXY was 7.75. The aqueous solubility of S-OXY was described as a function of pH and the free-base solubility. The mean partition coefficient log P was 3.33 using 1-octanol. The surface tensions of aqueous solutions of S-OXY decreased with increasing concentration, but no concentration-independent region was observed, indicating that S-OXY does notform micelles in aqueous solution. The dissolution rate of S-OXY from a compressed disk in 0.1 N HCl was rapid, whereas it was considerably slower at pH 7.4. Addition of 1% hexadecyltrimethylammonium bromide (CTAB) at pH 7.4 significantly improved the dissolution rate. S-OXY displayed very poor flow properties when compared to standard pharmaceutical excipients. XRPD results indicated that S-OXY exhibited a loss in crystallinity following ball milling. Hiestand tableting indices indicated that S-OXY has good bonding properties andforms strong compacts, but is likely to be susceptible to capping on ejection from the die. This indicated the needfor a plastically deformable excipient such as Avicel PH-101 in tablet formulations. PMID- 11411900 TI - Synthesis and properties of dextran-nalidixic acid ester as a colon-specific prodrug of nalidixic acid. AB - Dextran-nalidixic acid ester (dextran-NA) with a varied degree of substitution (DS) was synthesized as a colon-specific prodrug of nalidixic acid (NA). Solubility in water (mg/ml) of dextran-NA with DS (mg NA/100 mg dextran-NA) of 7, 19, or 32 was 57.57 (equivalent to 4.00 mg NA/ml), 0.53 (equivalent to 0.10 mg NA/ml), or 0.03 (equivalent to 0.01 mg NA/ml), respectively, and that for NA was 0.03 at 25 degrees C. To ensure the chemical stability of dextran-NA at conditions similar to those of the stomach and small intestine, dextran-NA was placed in a solution of pH 1.2 hydrochloric acid buffer or pH 6.8 phosphate buffer and incubated at 37 degrees C; no NA was detected during the 6 h of the incubation period, which indicated that dextran-NA might be chemically stable during the transit through the gastrointestinal tract. Degree of depolymerization (%) by dextranase determined by the 2,4-dinitrosalicylic acid (DNS) method at 37 degrees C for dextran-NA with DS of 7, 19, or 32 was 81, 68, or 8, respectively, in 8 h, and that for dextran was 91. When dextran-NA (equivalent to 50 microg of NA) with a DS of 7 or 17 was incubated with cecal contents (100 mg) of rats at 37 degrees C, the extent of NA released in 24 h was 41% or 32% of the dose, respectively. NA was not liberated from the incubation of dextran-NA with the homogenate of tissue and contents of the small intestine. PMID- 11411901 TI - Use of a moist granulation technique (MGT) to develop controlled-release dosage forms of acetaminophen. AB - The moist granulation technique (MGT), which involves agglomeration and moisture absorption, has only been applied to immediate-release dosage forms. Our results indicate that MGT appears to be applicable in developing a controlled-release formulation. A small amount of granulating fluid (water) was added to a powder blend to activate a dry binder (such as polyvinylpyrrolidone [PVP] at 2% and 3.6%) and to facilitate agglomeration. Then, a moisture-absorbing material (microcrystalline cellulose [MCC]) was added to absorb any excess moisture. By adding MCC in this way, a drying step was not necessary. Acetaminophen (APAP) was the model drug, with diluents lactose FastFlo and dicalcium phosphate. Hydroxypropylcellulose (HPC) was used as the controlled-release agent. The MGT was compared to conventional wet granulation (WG) and direct compression (DC) processing methods. The results indicate that MGT appears to be applicable in developing a controlled-release formulation. Particle size distribution of MGT and WG batches containing 3.6% PVP is similar. PMID- 11411902 TI - Effect of additives on stability of etoposide in PLGA microspheres. AB - The purpose of this article was to determine the shelf life of etoposide in poly(lactic-co-glycolic acid) (PLGA) microspheres prepared with and without additives (i.e., tricaprin and isopropyl myristic acid ester [IPM]). The microspheres were prepared by a single-emulsion solvent extraction technique with and without 25% w/w additive. The batches of microspheres were subjected to an accelerated stability study at two elevated temperatures (70 degrees C and 80 degrees C or 80 degrees C and 90 degrees C). Samples were taken at 7, 14, 21, 28, and 35 days for estimation of drug content by high-performance liquid chromatography (HPLC). The drug stability in the microspheres was determined by plotting the log percentage drug remaining versus time to obtain the degradation rate constant k of etoposide at the measured temperature. This degradation rate constant was then used in the Arrhenius equation to obtain the activation energy of etoposide, which was utilized to determine the shelf life of the microspheres at room temperature. The results showed that all three microsphere formulations had good long-term stability at room temperature (6.62-8.86 years at 25 degrees C). The plain microspheres were shown to possess a shelf life of 6.62 years, and the IPM and tricaprin were the most stable with shelf lives of 8.25 and 8.86 years, respectively. PMID- 11411903 TI - Poloxamer 407 as a thermogelling and adhesive polymer for rectal administration of short-chain fatty acids. AB - OBJECTIVES: The purpose of the study was to gel a rectal solution of short-chain fatty acids to decrease the loss of active materials in the colonic lumen and thereby optimize their absorption. METHODS: Five thermogels were prepared with poloxamer 407 at concentrations ranging from 17% to 20%. Their viscosities were measured at room temperature and 37 degrees C, and their gelling temperatures were determined. The adhesive properties of each gel were assessed in vitro at 37 degrees C. Short-chain fatty acid release was studied using Guyot cells. RESULTS: From the threshold concentration of 17.5%, the solutions, Newtonian at room temperature (50-80 mPa x s), gelled at 37 degrees C. The higher the concentration, the higher the viscosity (1750 to 49,000 mPa x s), the lower the gelling temperature (27.6 degrees C to 23.4 degrees C), and the stronger the work of adhesion (2.2 to 4.5 mJ). Short-chain fatty acid release from the 18% polymer gel was decreased by 60% compared to the rectal solution. CONCLUSION: The 18% poloxamer 407 concentration provided a solution that was liquid at room temperature, that gelled at 37 degrees C, possessed adhesive properties, and controlled short-chain fatty acid release. PMID- 11411904 TI - Molecular properties of propranolol hydrochloride prepared as drug-resin complexes. AB - Drug-resin complexes, as well as physical dispersions, containing varying contents of propranolol were prepared. The molecular properties of samples were investigated by differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), and infrared (IR) spectroscopy. In addition, the USP paddle method was used to determine the release behavior of drug from various formulations prepared from the samples. The data from DSC and XRPD indicated that the molecular state of drug in the complexes was amorphous, whereas that in the physical dispersions exhibited the crystalline state of pure drug. These results suggested that the molecule of drug prepared as drug-resin complexes was monomolecularly dispersed in the resin bead. The IR study provided evidence that demonstrated the interaction between the drug and resin in the complexes. The release behavior of drug from the complexes was governed by the cross-linkage structure and equilibrium treatment of drug exchange of resin. PMID- 11411905 TI - Effects of low-viscosity sodium hyaluronate preparation on the pulmonary absorption of rh-insulin in rats. AB - PURPOSE: A low-viscosity formulation for pulmonary delivery of rh-insulin as model peptide drugs was developed using a solution of sodium hyaluronate. METHOD: The effects of different concentrations and pH values of low-viscosity solutions of hyaluronate on the pulmonary absorption of rh-insulin were examined after intratracheal administration in rats. The permeation of fluorescein isothiocyanate (FITC)-dextran (molecular weight 4300; FD-4) and insulin through excised rat trachea in vitro were also examined. RESULTS: The hyaluronate (2140 kDa) solutions (0.1% and 0.2% w/v) at pH 7.0 significantly enhanced the pharmacological availability (PAB) of insulin compared to the aqueous solution of insulin at pH 7.0. The absorption-enhancing effect at a concentration of 0.1% w/v hyaluronate was greater than that at a concentration of 0.2% w/v hyaluronate. Furthermore, the greatest absorption-enhancing effect was obtained, regardless of the molecular weight of hyaluronate, when the concentration of hyaluronate was adjusted to 0.47 microM. Absorption-enhancing effects were consistent with the effect of a 0.1 w/v hyaluronate preparation at pH 4.0 and 7.0 on the permeation of FITC-dextran and insulin through excised rat trachea in vitro. CONCLUSION: Low viscosity hyaluronate preparation was shown to be a useful vehicle for pulmonary delivery of peptide drugs. PMID- 11411906 TI - A double-blind investigation of the potential systemic absorption of isotretinoin, when combined with chemical sunscreens, following topical application to patients with widespread acne of the face and trunk. AB - This study assessed the systemic absorption of isotretinoin and its metabolites, during a 4-week application of a cream containing 0.1% isotretinoin and chemical sunscreens, compared with a 4% benzoyl peroxide cream, in patients with acne on the face and trunk. Blood was drawn at weeks 0, 1, 2, 3 and 4 and at 96 h post treatment. Plasma levels of isotretinoin (13-cis-retinoic acid) and tretinoin (all-trans-retinoic acid) were quantified by liquid chromatography with tandem mass spectrometry and the presence of their combined 4-oxo metabolites were monitored from the peak area ratios observed. The isotretinoin group showed no statistically or clinically significant increases in plasma retinoid levels and mean levels did not exceed +/-2 SD of the mean pre-treatment values, indicating that endogenous levels were not being exceeded. No significant differences were detected between the isotretinoin group and the 4% benzoyl peroxide group. These findings indicated that retinoid absorption from a cream containing 0.1% isotretinoin and chemical sunscreens was clinically insignificant, when applied to patients with widespread acne. PMID- 11411907 TI - Tumour necrosis factor-alpha promoter polymorphism in erythema nodosum. AB - Erythema nodosum is a common skin disease characterized by erythematous, tender subcutaneous nodules, mostly located on the lower extremities. Little is known about its pathogenesis, although a wide variety of aetiological factors (e.g. bacterial and viral infections, neoplastic diseases and drugs) have been described. Sarcoidosis, a typical granulomatous disease, often occurs in association with erythema nodosum (Loefgren syndrome). Since granulomatous diseases have been closely linked to a deregulated tumour necrosis factor (TNF) alpha production, it was tempting to speculate whether TNF-alpha might play a role in the pathogenesis of erythema nodosum, at least in cases associated with sarcoidosis. A previously described nucleotide exchange, (G-A) at position -308 in the human TNF-alpha gene promoter, has been shown to be a major cause for enhanced TNF-alpha production. In the present report, we investigated the genomic TNF-alpha promoter region in patients suffering from EN with and without underlying sarcoidosis. Our results showed a strong correlation between the uncommon TNF A II allele and sarcoidosis-associated erythema nodosum. Patients with erythema nodosum without underlying sarcoidosis displayed a similar allele frequency compared with controls. Taken together, we provide evidence that erythema nodosum in association with sarcoidosis might be pathogenically linked to altered TNF-alpha production due to a genetic promoter polymorphism. PMID- 11411908 TI - Comparison of cyclosporin and UVAB phototherapy for intermittent one-year treatment of atopic dermatitis. AB - Although cyclosporin is effective for the treatment of severe atopic dermatitis, phototherapy is the standard second-line treatment for this disease. An open, randomized, controlled, parallel-group study was conducted to compare the efficacy, influence on quality of life and safety of cyclosporin and UVAB phototherapy during 1 year of intermittent treatment of atopic dermatitis in adult patients. The main endpoints of the study were the number of days in remission and the influence on quality of life. Seventy-two patients were treated, 36 in each group. Cyclosporin produced significantly more days in remission than UVAB phototherapy during the 1-year study period. At the end of the study no difference between the 2 groups was noted in terms of quality of life. A significant increase in serum creatinine occurred in 2 patients and 7 patients developed mild or moderate hypertension during cyclosporin treatment. It can be concluded that intermittent cyclosporin seems to be more effective than UVAB and is reasonably safe for the treatment of atopic dermatitis over a 1-year treatment period. PMID- 11411909 TI - No human leukocyte antigen-A, -B or -DR association in Swedish patients with hidradenitis suppurativa. AB - Hidradenitis suppurativa (HS) is a cicatrizing, inflammatory and recurrent disease restricted to inverse skin, such as that of the axilla and groin of younger adults. In a previous study, using serological tissue-typing techniques, no significant increases in the human leukocyte antigen (HLA)-A and -B specificities were found in patients with HS. The aim of this study was to determine the frequencies of HLA-A, -B and, for the first time, HLA-DR alleles, using genomic tissue-typing methods in patients with HS. Forty-two unrelated Swedish patients with HS were included and compared with 250 controls. According to clinical staging adopted from Hurley all of the patients had stage II HS, i.e. recurrent abscesses with tract formation and cicatrization and single or multiple widely separated lesions. No association with HLA-A, -B or -DRB1 alleles was found in patients with HS. Genetic factors associated with the HLA class I or II regions do not appear to contribute significantly to the possible genetic susceptibility of HS. PMID- 11411910 TI - Cytokine expression of skin T-lymphocytes from patients with atopic dermatitis. AB - We analysed the cytokine profile of skin T cells by establishing 11 T-cell lines from adult patients with moderate-to-severe atopic eczema using T-cell growth factors interleukin-2 and interleukin-4. We compared T-cell lines from lesional skin of atopic dermatitis patients with those from non-atopic skin of patients with other skin diseases, observing that T-cell lines of patients with atopic dermatitis unstimulated cultures expressed a Th1 profile. After stimulation with anti-CD3 and anti-CD28 monoclonal antibodies, the cytokine expression showed rapid initial upregulation of Th2 followed by a Th1 profile. Furthermore, strong upregulation of interleukin-10 was observed after 24 h stimulation. Our findings suggest that skin T-lymphocytes from atopic dermatitis patients seem to consist of a heterogenous population of Th1 and Th2 or Th0 cells and the results for secreted cytokines indicate that T-cell lines from each inflammatory skin disease showed the corresponding disease-specific original cytokine profile. PMID- 11411911 TI - Allergic contact sensitization in an adult Danish population: two cross-sectional surveys eight years apart (the Copenhagen Allergy Study). AB - In 1990 and 1998 15-41-year-old people were patch-tested in 2 cross-sectional studies of random samples of the population in the western part of Copenhagen County, Denmark. In 1990, 290 subjects and in 1998, 469 subjects were patch tested. The participation rates were 69% and 51%, respectively. Contact sensitivity to one or more haptens was found in 15.9% and 18.6% in 1990 and 1998, respectively. Nickel sensitivity is still the most common contact sensitivity. The risk of contact sensitivity to the cosmetic-related haptens included in the series (formaldehyde was not included) increased significantly from 2.4% in 1990 to 5.8% in 1998 (odds ratio 2.44, 95% confidence interval 1.04-5.73). The prevalence of contact sensitivity to cosmetic-related allergens has been doubled between 1990 and 1998. PMID- 11411912 TI - Recurrent genital herpes in a population attending a clinic for sexually transmitted diseases. AB - Patients with recurrent genital herpes attending a sexually transmitted disease clinic were studied and transmission of the infection was elucidated by evaluating serostatus in their partners. Of 84 patients attending for recurrent genital herpes, 94% had a herpes simplex virus type 2 (HSV-2) infection and only 6% (5 patients) a type 1 infection. The mean age of the patients was 36 years and the duration of their infection was up to 37 years (median 4 years). In most patients the number of recurrences had not decreased between the first year and the last year. About half had experienced a more severe first episode infection. Of the patients, 64% were not aware of asymptomatic shedding and the risk of sexual transmission without clinical symptoms. Of 67 steady partners of patients with genital HSV-2, 15% had a history of genital herpes. By HSV serology, HSV-2 antibodies (indicating subclinical genital herpes) were demonstrated in more than half of the partners. The duration of the relationship or condom use did not seem to influence the frequency of transmission to the partner, which may indicate an individual susceptibility for acquiring a genital HSV-2 infection. Eleven per cent of the patients were on suppressive antiviral therapy, while 39% had no experience of antiviral therapy. Type-specific HSV serology was found to be of value in counselling partners of patients with genital herpes. PMID- 11411913 TI - Dermatoscopic prediction of melanoma thickness using latent trait analysis and likelihood ratios. AB - Breslow thickness and Clark level can be used to determine surgical procedures for cutaneous malignant melanoma and patient eligibility for experimental adjuvant therapy. Efforts to predict the thickness of melanomas using dermatoscopy have focused on differences between single dermatoscopic findings. The aim of this study was to develop a method for preoperative identification of melanomas of > or = 1 mm Breslow thickness using the entire range of dermatoscopic findings. Sixty-five melanomas were assessed for the presence of 22 dermatoscopic features. Ten dermatoscopic features showed differences in thick and thin melanomas and were selected for further analysis. A latent trait analysis construct implied that a progression in dermatoscopic features was associated with advancement of melanomas. Early melanomas are characterized by a light brown colour, a pigment network and irregularity or heterogeneity. Gray blue areas, white scar-like areas and an atypical vascular pattern gradually displace these features. Likelihood ratios were determined for these 6 dermatoscopic findings and an algorithm for calculating the probability of thick malignant melanoma was established. PMID- 11411914 TI - Granulomatous slack skin: a distinct disorder or a variant of mycosis fungoides? AB - About 75% of cutaneous lymphomas belong to the group of T-cell lymphomas. Mycosis fungoides is the most common entity in this group. Granulomatous slack skin is a rare form of cutaneous T-cell lymphoma closely related to mycosis fungoides. We present here a patient with areas of lax skin for several years who developed a generalized erythroderma with associated immunoactivation and a deterioration in his general condition. This report discusses clinically and histologically the differential diagnoses, namely granulomatous slack skin and granulomatous mycosis fungoides, and suggests that these 2 disorders are only variants in the broad spectrum of a single disease. PMID- 11411915 TI - Aleukemic leukemia cutis presenting as benign-appearing exanthema. AB - Aleukemic leukemia cutis is a rare condition characterized by the infiltration of the skin by leukemic cells before their appearance in the peripheral blood or bone marrow. We report here a 62-year-old seemingly healthy patient who presented with disseminated erythematous maculae. A skin biopsy showed leukemia cutis of monocytic type. No involvement of bone marrow or peripheral blood was found. The patient developed acute monocytic leukemia 7 months later. We present this case to illustrate how leukemia cutis can masquerade as a clinically benign-appearing cutaneous eruption without leukemic changes in blood or bone marrow. To confirm the diagnosis of aleukemic leukemia cutis, immunohistochemistry of the skin lesions as well as a complete staging procedure is necessary. PMID- 11411916 TI - An unusual form of generalized granuloma annulare in a patient with insulin dependent diabetes mellitus. AB - The generalized form of granuloma annulare may be associated with systemic disorders, including diabetes mellitus. We describe here an unusual form of generalized granuloma annulare in a patient with complicated insulin-dependent diabetes mellitus. The cutaneous eruption had been present for decades as non pruritic, persistent, violet-brown patches with raised edges. There were flexion deformities of the small joints of the hands and feet associated with thickening of the skin over dorsa of the fingers. The patient is currently on isotretinoin therapy, with partial resolution of lesions at 3 months follow-up. PMID- 11411917 TI - Chronic infection due to Fusarium oxysporum mimicking lupus vulgaris: case report and review of cutaneous involvement in fusariosis. AB - A 67-year-old female presented with a 20-year-old lesion involving the right ear and preauricular area mimicking tuberculous lupus. Fusarium oxysporum infection was confirmed by biopsy studies and cultures. The biopsy specimen showed an unusually extensive dermal invasion with fungal hyphae. This is an uncommon clinical presentation for Fusarium infection in a healthy patient. When referred to us, the patient had received antifungal therapy with itraconazole without any benefit. Improvement was obtained with fluconazole therapy. The spectrum of cutaneous involvement related to Fusarium spp. includes toxic reactions, colonization, superficial indolent infection, deep cutaneous or subcutaneous infections and disseminated infection. PMID- 11411918 TI - Characterization of a novel point mutation (Arg432His) in X-linked ichthyosis. PMID- 11411919 TI - Infiltration of activated eosinophils in the skin lesions of atopic dermatitis. PMID- 11411920 TI - Acral Darier's disease successfully treated with adapalene. PMID- 11411921 TI - Famciclovir in treatment of acute herpes zoster: results of two post-marketing surveillance studies in Germany. PMID- 11411922 TI - Oral terbinafine for tinea capitis due to Microsporum canis in children. PMID- 11411923 TI - IgE-mediated urticaria/angioedema after ingestion of mussels. PMID- 11411924 TI - Systemic sclerosis following physical trauma. PMID- 11411925 TI - Disseminated superficial porokeratosis with mucosal involvement. PMID- 11411926 TI - Mechanic's hand: a clinical diagnostic aid in dermatomyositis. PMID- 11411927 TI - Lichen myxedematosus associated with chronic hepatitis C: a case report. PMID- 11411928 TI - The Asp84Glu variant of the MC1R gene in Norwegian melanoma patients. PMID- 11411929 TI - Erythroderma due to dermatophyte. PMID- 11411930 TI - Successful sulfasalazine treatment of severe chronic idiopathic urticaria associated with pressure urticaria. PMID- 11411931 TI - Epidermolysis bullosa acquisita treated with basiliximab, an interleukin-2 receptor antibody. PMID- 11411932 TI - Capnocytophaga canimorsus sepsis causing Sweet's syndrome. PMID- 11411933 TI - Sarcoidosis developing after stopping oral prednisolone for systemic lupus erythematosus. PMID- 11411934 TI - Darier's disease seen with cutis verticis gyrata. PMID- 11411935 TI - Successful treatment of extensive condyloma acuminata of the inguinal area and thigh with topical imiquimod cream. PMID- 11411936 TI - Unusually large cutaneous metastases of renal cell carcinoma. PMID- 11411937 TI - Occurrence of neutrophils and activated Th1 cells in UVB-induced erythema. AB - We investigated the sequential changes in infiltrating inflammatory cells and several cytokine levels over a period of 48 h in human back skin exposed to 3 minimal erythematous doses of UVB. The measurement of blood flow, using a laser Doppler method, indicated that UVB-induced erythema reached a peak 12-24 h after irradiation. Immunohistochemically, an increase in the number of CD4+ T cells was observed in perivascular areas 6 h after the UVB treatment and continued for up to 48 h. CD8+ T cells were scarce until 24 h, but their numbers gradually increased thereafter. HLA-DR+ cells were detected perivascularly and interstitially in parallel with the pattern of CD4+ T-cell infiltration. In contrast, neutrophils were found 3 h after UVB exposure and reached a peak at 24 h. Using a RT-PCR method, we demonstrated that mRNAs for the Th1 cytokines (interferon-gamma and interleukin-2), together with a proinflammatory cytokine (interleukin-8), became detectable at 6 h, whereas mRNA for the Th2 cytokine (interleukin-4) was not found at all during the first 48 h. In contrast, we found an increase in mRNA levels for C3 and tumor necrosis factor-alpha even at 3 h, suggesting a relationship between complement activation and accumulating neutrophils. Our results suggest that neutrophils and CD4+ T cells in UVB-induced inflammation play different roles: neutrophils are more closely related to UVB induced erythema, while T cells appear to be involved in subsequent dermal and epidermal inflammation accompanied by epidermal hyperproliferation. PMID- 11411938 TI - Evidence for the mitochondrial origin of the eye lenses in embryos of Entobdella soleae (Plathelminthes, Monogenea). AB - The lens associated with each of the four pigmented eyes of the oncomiracidium of Entobdella soleae (Plathelminthes, Monogenea, Capsalidae) develops in a special region of the pigment cup cell of the eye. It is confirmed that the inner of the two membranes enclosing each lens bears short, inwardly projecting, membranous profiles identical to mitochondrial cristae. Studies of embryos incubated for 19 days at 12 degrees C (hatching begins at 28 days at this temperature) revealed that the matrix of the developing lens of each anterior eye contains many mitochondrial membrane compartments, some having the configuration of separate, small mitochondria. The implication is that the lens is derived from many fused mitochondria, rather than from a single large one. The anterior eyes of 19-day old embryos are less well developed than the posterior eyes. Pigment granules in the anterior eyes appear to be at the premelanosome stage and contain dispersed dense particles lacking an obvious orderly arrangement. The posterior eyes mostly contain mature melanosomes. Membranous compartments in the matrix of the posterior eye lenses are rare. Apart from longer peripheral cristae, lenses of 22 day-old embryos are identical with those of oncomiracidia. The evolution of mitochondrial lenses in Plathelminthes is considered. PMID- 11411939 TI - Viability and infectivity of oocysts recovered from clams, Ruditapes philippinarum, experimentally contaminated with Cryptosporidium parvum. AB - This study confirms the important role of marine bivalve molluscs, destined for human consumption, as transmitters of cryptosporidiosis, zoonotic diarrhoeal disease caused by Cryptosporidium parvum. C. parvum oocysts recovered from seawater clams (Ruditapes philippinarum) were viable and infective in five of eight infected neonatal CD-1 Swiss mice. Oocysts were observed in clam gill and gastrointestinal tract tissue homogenates as well as in gill histological sections, by an immunofluorescent antibody technique. In vitro viability of recovered oocysts was also determined using fluorogenic vital dyes (75% viability). PMID- 11411940 TI - Physical and transcriptional organization of the ribosomal RNA genes of the savannah-type Trypanosoma congolense. AB - Ribosomal RNA genes have been cloned from the major species of African trypanosomes. Complete nucleotide sequence composition of the small subunit (SSU) and portions of the large subunit (LSU) ribosomal RNA genes was determined for each of these trypanosome species. In contrast to the situation in Trypanosoma brucei, in savannah-type T. congolense the LSU ribosomal RNA is cleaved twice, to generate two additional prominent fragments. This leads to the different profiles observed when the rRNA molecules from these two trypanosome species are resolved in agarose gels. From the nucleotide sequences of the 18S RNA, a phylogenetic tree was derived depicting the relationships among the T. congolense complex of trypanosomes and the other species of trypanosomes. PMID- 11411941 TI - Morphological and genetic differentiation of Rodentolepis straminea (Goeze, 1752) and Rodentolepis microstoma (Dujardin, 1845) (Hymenolepididae). AB - The two related species, Rodentolepis straminea (Goeze, 1782) and Rodentolepis microstoma (Dujardin, 1845) (Cestoda, Hymenolepididae), both parasites of rodents, were compared morphologically and electrophoretically. Adult worms were isolated from three wild rodent species of the family Muridae (Apodemus flavicollis, Apodemus sylvaticus, and Mus musculus) from three different sites in Spain and France. Although these two species were strikingly similar in morphological appearance, some of the morphological and metrical features analysed (scolex, mature segments and eggs) can be used for differentiation. Fixed allelic differences were found. Of the ten enzymes detected by starch-gel electrophoresis, six (AAT, AK, GPI, MDH, NP, PGM) showed characteristic isoenzyme profiles in each species. Only in MPI, PEPC, PEPD, and ME enzyme loci were no differences found. The study revealed that the two taxa can be clearly differentiated. PMID- 11411942 TI - Evaluation of rapid immunocapture assays for diagnosis of Plasmodium vivax in Korea. AB - The rapid immunocapture assays, OptiMal and ICT, were evaluated from 87 individuals for the diagnosis of malaria infections directly from whole blood. A total of 87 individuals was examined for malaria parasites by microscopic examination of Giemsa-stained blood smears, and 65 cases were positive for Plasmodium vivax by microscopy. Correspondingly, the OptiMal test identified malaria infection in 45 cases (69.2%) of microscopy positive cases. Of these, two cases were misinterpreted as Plasmodium falciparum, whereas ICT detected P. vivax infection in 29 (44.6%) patients. We would like to propose that rapid immuno capture assays are an easy method that can serve as a useful tool in addition to microscopy for the diagnosis of malaria, but sensitivity is not yet satisfactory for diagnosis of P. vivax in Korea. PMID- 11411943 TI - Inhibition of Cryptosporidium infection in mice treated with a cyclodextrin inclusion complex with diloxanide furoate. AB - The efficacies of diloxanide furoate, beta-cyclodextrin and a cyclodextrin inclusion complex against Cryptosporidium parvum were evaluated in a suckling murine model. Efficacy was established by numbers of oocysts recovered from the intestinal tract of mice on day 7 postinfection. The level of infection in treated mice was significantly lower than in control mice and, surprisingly, the most efficacious treatment was beta-cyclodextrin, an excipient used in pharmaceutical technology. PMID- 11411944 TI - HSP60, HSP70 and HSP90 from Trichinella spiralis as targets of humoral immune response in rats. AB - This study identifies three heat shock proteins (HSPs) using purified preparations from Trichinella spiralis larvae. The proteins: HSP60, HSP70 and HSP90 were found to be targets of the humoral immune response in rats. Three approaches were adopted to obtain T. spiralis HSP-enriched material and/or to purify HSPs to homogeneity. The former product was prepared using affinity chromatography on gelatin sepharose and elution with ATP. Pure 90 kDa-protein was isolated from parasite extract by sequential DEAE (A50) column chromatograpy and preparative electrophoresis. Immunoblot analysis using monoclonal antibodies to HSP60, HSP70 and HSP90 detected the HSP60 and HSP70 in the affinity-purified product and HSP90 in the product obtained by sequential anionic chromatography and preparative electrophoresis. Finally, the reactivy of preimmune, T. spiralis immune and irrelevant immune rat sera on immunoblots were also examined. Only sera taken from infected rats at time-points after day 7 following the first infection exhibited activity against 60, 70 and 90 kDa proteins on blots. The fact that the serum antibodies were anti-HSP was established by immunoadsorption of HSPs to microtiter plates coated with anti-HSP60, anti-HSP70, or anti-HSP90 and using rat sera, positive on blots, to also give positive scores by continued enzyme-linked immunosorbent assay. PMID- 11411945 TI - Eimeria parasites of domestic fowl: genetic relationships of different isolates estimated from random amplified polymorphic DNA. AB - Genetic relationships between Eimeria parasites of domestic fowl, including three isolates of E. acervulina, one of E. maxima, four of E. mitis, six of E. praecox, one of E. tenella, and two of uncertain identity, were analyzed by random amplified polymorphic DNA (RAPD) and the unweighted pair-group method with arithmetic mean, using 12 primers. Each primer amplified 4-34 DNA fragments/isolate. The similarity coefficients and phenograms were calculated from RAPD products with 230-2,000 bp in silver-stained polyacrylamide gels. Some primers generated RAPD markers which were species-specific for E. acervulina, E. mitis, and E. praecox. The phenograms revealed six clusters, each corresponding to an individual Eimeria species. The pBP and pBP2 isolates, of uncertain identity, clustered with the E. acervulina isolates (100% bootstrap). The intraspecific relationships showed certain degree of genetic isolation of the Eimeria populations, and it was associated with broiler house and with geographic separation. PMID- 11411946 TI - Ultrastructure of the acanthella of Paratenuisentis ambiguus (Acanthocephala). AB - The fine structure of the early acanthella of Paratenuisentis ambiguus (Eoacanthocephala) was investigated. This developmental stage is characterised by losing its ability to move and by differentiation of adult structures. The frontal syncytium, present in the first developmental stage (the acanthor), is lost, while the epidermis and central syncytium persist. The epidermis of the acanthella contains a number of giant nuclei that are arranged into several small groups. The central syncytium is subdivided into different masses, containing nuclei that will give rise to the organs of the adult. The 'uncinogenous bands' extend into the anterior body of the acanthella. Formation of the hooks takes place within these strands. In all investigated stages no extracellular materials were observed. Posterior of the uncinogenous bands lies the brain anlage and the primordia of the reproductive system. Neither a sense organ nor a nervous system were found. PMID- 11411947 TI - Evaluation of immunomagnetic separation method for detection of Giardia for different reaction times and reaction volumes. AB - Immunomagnetic separation (IMS) has been specified as a standard method for the measurement of Giardia. In this study, Dynal IMS was evaluated on the basis of recovery efficiencies of Giardia cysts for various IMS operational conditions. The average recoveries for Giardia in deionized, treated and raw water samples were 82.6 +/- 12.2% (n = 6), 75.6 +/- 15.2% (n = 3), and 70.6 +/- 18.2% (n = 3), respectively. Significant changes in recovery were observed by altering the debris ratio and the debris components of water samples. Changing the reaction volume within the same vessel had no significant effect on cyst recovery efficiencies. However, prolonging the reaction time did increase recovery efficiencies. PMID- 11411948 TI - The definitive and intermediate hosts of Fasciola hepatica in the natural watercress beds in central France. AB - Field investigations were carried out over a two-year period in 52 natural watercress beds located in the Limousin region of central France to list the mammal and bird species that frequented these sites. This enabled detection of the definitive hosts of Fasciola hepatica and determination of the prevalence of natural infection in snails. A total of 13 mammal and five bird species were listed in these watercress beds. Adult flukes were found in Lepus capensis (39.2%), Oryctolagus cuniculus (42.0%), and Sylvilagus floridanus (25.0%). No infection with F. hepatica was noted in the five species of rodents studied. Snails infected with F. hepatica were found in 14 watercress beds. The global prevalence of natural infection was 1.1% in Lymnaea truncatula and 0.3% in L. glabra. Among the other trematode larval forms detected, the most frequent was Haplometra cylindracea (0.5%). In the Limousin region, the presence of hares and rabbits in watercress beds ensured the continuation of the F. hepatica life cycle and permitted the subsequent infection of humans when this wild watercress was eaten. PMID- 11411949 TI - The karyotype of Trichobilharzia regenti Horak, Kolarova et Dvorak, 1998 (Digenea: Schistosomatidae), a nasal avian schistome in Central Europe. AB - The karyotype of a European bird nasal schistosome, Trichobilharzia regenti Horak, Kolarova et Dvofak, 1998, was determined. The chromosome set comprises eight pairs, seven of which are autosomes and one sex pair consisting of ZZ in males and ZW in females. The Z is the largest chromosome of the set, having a submeta-metacentric location of centromere, while the W is a small metacentric chromosome. More than 60% of the analyzed cells originating from both male and female sporocysts contained one or two small submetacentric supernumerary B chromosomes, occurring more frequently in female than in male cells. The formula of the karyotype structure is 2n = 16 + (B), n=5m+1sm-m+ 1sm+Zsm-m/Wm+(Bsm). The C-banding method revealed small pericentric heterochromatin blocks in the sex chromosome Z and all autosomes, except pairs 5 and 6. Satellited pair 5 showed a distinct terminal band representing the nucleolar organizer. Both the W and B chromosomes contained conspicuous heterochromatin blocks at the long arm adjacent to the centromere. The currently described supernumerary chromosomes in T. regenti represent the first record of B chromosomes within the family Schistosomatidae. PMID- 11411950 TI - The guild as a concept and a means in ecological parasitology. AB - The use of the term guild community as an intermediate level between component and supra-community proves to be advantageous within the different steps of parasite communities. It can comprise more information than a component community but researchers are able to analyse important parts of the supra-community without being forced to include all parasites in an ecosystem. As an example, the infestation of fishes of four families (= four guilds) from the Salzhaff (NW Mecklenburg) by several parasite specialists and generalists is analysed. Thus, processes of despecialisation in an extreme environment like brackish water can be demonstrated in an adequate manner. PMID- 11411951 TI - Differential display analysis of hemocytes from schistosome-resistant and schistosome-susceptible intermediate hosts. AB - Hemocytes from schistosome-resistant and schistosome-susceptible Biomphalaria glabrata differ fundamentally in their behavior towards an invading parasite. When the schistosome infects a resistant snail host it is quickly surrounded by hemocytes, encapsulated and destroyed. Hemocytes from susceptible hosts fail to kill the parasite. To detect the differences between these two host phenotypes, we used differential-display reverse-transcription PCR (DDRT-PCR), based on RNA extracted from isolated hemocytes. A number of differentially expressed fragments from resistant and susceptible snails were detected by DDRT-PCR and confirmed using single-strand conformation polymorphism. These methods proved to be sensitive enough to allow comparison and verification of differential gene expression in our system, where only small numbers of cells are available. The most interesting phenotype-specific fragments detected so far show sequence homologies to an adhesion molecule, defensin, serine/ threonine kinases, peroxidases and glycosidases. PMID- 11411952 TI - Anisakis infestation in marine fish and cephalopods from Galician waters: an updated perspective. AB - A total of 2,673 fresh specimens of cephalopod and fish representing 35 species were obtained from commercial local fisheries in Galician waters (NW Spain). They were examined for anisakid nematodes by digestion of the muscle and elution of the viscera and whole body cavity. All larval nematodes recovered were identified by light microscopy and multilocus electrophoresis as belonging to the species Anisakis simplex sensu stricto and A. pegreffii. Encysted larvae mostly occurred in the viscera but were also found in the flesh of squid and fish. Demographic values for larval nematodes are discussed in relation to host preferences and the ecological niche of both anisakid species at the sampling area. Primary recommendations are also expressed concerning the effects of current fishing and aquaculture practices on the Anisakis problem. PMID- 11411953 TI - Serological response to Cryptosporidium parvum in adult cattle from the Andean region of Colombia. AB - Single faecal and serum samples were individually collected from 135 asymptomatic adult cows on seven farms in Cundinamarca (Colombian Andean region). Tests for the presence of oocysts of Cryptosporidium parvum (carbol fuchsin stain) and Eimeria spp (flotation in saturated saline solution) revealed that none of the animals had coccidia in their faeces. The IgG antibody levels to C. parvum were measured by an enzyme-linked immunosorbent assay (ELISA) technique and the reactivity to C. parvum antigens by a Western blotting procedure. Cryptosporidial antibodies were detected in cattle from all farms, with 53.3% (72 animals) being seropositive. Sera recognized 5-11 protein fractions with molecular masses ranging from 12 14 kDa to 97-100 kDa. Sera considered as positive by ELISA reacted intensely and more frequently with protein fractions of approximately 20 22, 42-48, 51-57 and 60-69 kDa, whereas only the 42-48 kDa antigen was strongly recognized by sera without IgG antibodies. The presence of IgG antibody against C. parvum in most animals, as well as the reactivity to major proteins of C. parvum, could be indicative of continuous exposure to this parasite. PMID- 11411954 TI - Interaction of Trypanosoma rangeli Tejera, 1920 with different cell lines in vitro. AB - Intracellular multiplication of Trypanosoma rangeli was evaluated in vitro using experimental infection of Vero cells, murine macrophages, and promonocytes with T. rangeli Choachi, Macias, and SC-58 clone B1 strains. Our results revealed a low infectivity of all T. rangeli strains to these cell lines. Macrophages showed the highest infection rate; however, intracellular forms were no longer observed 48 h post infection Despite the observation of intracellular parasites up to 144 h post infection, the infection rates of Vero cells and J774G.8 promonocytes with these parasite strains were always below 5%. Pre-incubation of parasites with normal mouse serum increased the initial infectivity but not the time course of the infection. Under our experimental conditions, we did not observe any evidence of intracellular multiplication of T. rangeli within these cell lines. PMID- 11411955 TI - Chemoprophylactic praziquantel treatment in experimental hydatidosis. AB - In the present study, new data on the chemoprophylactic treatment of experimental hydatid disease with praziquantel are presented. In vivo studies have been performed reducing the treatment to 1 and 2 months. NMRI female mice inoculated with protoscolices of Echinococcus granulosus were treated with praziquantel 48 h p.i. at a dose of 600 mg/kg, 5 days a week for 1 or 2 months. After 4 months p.i. the animals were killed, and the cysts obtained were counted, weighed and processed for observations by means of transmission electron microscopy. Both wet weight and number of hyaline cysts found in treated mice were significantly lower than those found in control groups in both experiments (1 and 2 months of treatment), obtaining a high efficacy in all cases. Moreover, severe ultrastructural damage in the treated cysts was seen compared to the control cysts. The reduction in treatment time lays the basis for an important advance in the control and prevention of hydatidosis. PMID- 11411956 TI - Silencing of CD21 expression in synovial lymphocytes is independent of methylation of the CD21 promoter CpG island. AB - The complement receptor II (CD21) recognises the complement component C3d of immune complexes. Expression of the CD21 gene is tightly regulated during B lymphocyte differentiation. Only mature B lymphocytes express CD21 but not pro-, pre-, or plasma B lymphocytes. Previously we found that pro-, pre-, and intermediate B lymphocytes contain a methylated CpG island and do not express CD21. CD21-expressing mature B lymphocytes, plasma B lymphocytes, and nonlymphoid cells carried a demethylated CD21 CpG island. Furthermore, we found that synovial lymphocytes from patients with rheumatic disease show reduced expression of CD21. This observation tempted us to analyse the methylation status of the CD21 CpG island in peripheral blood mononuclear cells and synovial fluid mononuclear cells derived from these patients. While methylation is involved in silencing CD21 in early types of B lymphocytes, we found the CD21-CpG island to be demethylated in peripheral blood mononuclear cells and synovial fluid mononuclear cells of patient DNA. PMID- 11411957 TI - Treatment of rheumatoid arthritis by oral administration of bovine tracheal type II collagen. AB - We evaluated the efficacy and safety of orally administered bovine tracheal type II collagen (CGII) in the treatment of rheumatoid arthritis (RA). Twenty RA patients received 0.5 mg/day of CGII for 12 weeks. Eighteen of them had improvements in the clinical parameters studied (swollen and tender joint counts, 15-m walking time, duration of morning stiffness, and physician's global assessment of disease activity). Anti-CGII antibodies were positive in 57% and rheumatoid factor (RF) in 71% of the patients with a short history of RA ( < or =2 years), whereas only 23% of those with long histories (>2 years) presented autoantibodies to CGII and 38% had positive RF. After the treatment, four patients showed reduced RF levels and all those with detectable serum tumor necrosis factor alpha (TNF-alpha) experienced its return to normal or levels below those at study entry. Although a placebo effect cannot be discounted, the oral administration of bovine tracheal CGII induced clinical benefits in 90% of the patients, without the side effects usually associated with treatment. This is the first study showing that feeding CGII can induce reductions in RF and TNF alpha. The data justify further controlled studies to assess the long-term efficacy of this treatment approach. PMID- 11411958 TI - Arthritis in familial Mediterranean fever. AB - We studied the rheumatic and various clinical manifestations of familial Mediterranean fever (FMF) in Lebanon. A retrospective review was performed of the medical records of 74 FMF patients seen at the American University of Beirut Medical Centre (AUB-MC) from 1979 to 1996. We also reviewed the medical literature from 1968 to 2000 using MEDLINE and the key words "familial Mediterranean fever" and "arthritis". Arthritis was the presenting symptom in 12 cases (16.2%). Twenty-three patients (31%) had definite arthritis during the course of the disease that was monoarticular in 16 (70%), oligoarticular in six (26%), and polyarticular (rheumatoid-like) in one (4%). Arthritis of the large joints of the knees and ankles was the most frequent articular involvement. The arthritis was transient, monoarticular, nonerosive, and nondeforming in the majority of cases. Four patients (5.4%) had chronic arthritis, with one requiring total hip replacement. As in previous reports on arthritis of FMF, the majority of FMF patients studied in Lebanon had a transient monoarticular nonerosive and nondeforming type of arthritis affecting predominantly the large joints of the lower extremities. PMID- 11411959 TI - The new antirheumatic drug KE-298 suppresses monocyte chemoattractant protein (MCP)-1 and RANTES production in rats with adjuvant-induced arthritis and in IL 1beta-stimulated synoviocytes of patients with rheumatoid arthritis. AB - We analyzed the effects of the new antirheumatic drug KE-298 on monocyte chemoattractant protein (MCP)-1 and regulated on activation normal T-cell expressed and secreted (RANTES) production in rats with adjuvant-induced arthritis and in interleukin (IL)-1beta-stimulated rheumatoid arthritis (RA) synoviocytes. In rats with adjuvant-induced arthritis, the enhanced production of MCP-1 and RANTES and the development of arthritis were suppressed by oral treatment with 100 mg/kg per day of KE-298 for 18 days. Furthermore, KE-298 (10 100 microg/ml) suppressed MCP-1 and RANTES production by IL-1beta-stimulated RA synoviocytes through inhibition of NF-kappaB and AP-1 activation. These results suggest that the inhibitory effect of KE-298 on MCP-1 and RANTES production might partly explain its efficacy in rats with adjuvant-induced arthritis and in patients with RA. PMID- 11411960 TI - Elevated levels of nitrate in rheumatoid arthritis. AB - Nitric oxide (NO) is a free radical that plays important roles in many physiological and pathological processes. Evidence suggests that NO participates in the pathogenesis of inflammatory reactions in many autoimmune and inflammatory diseases such as rheumatoid arthritis (RA). The purpose of this study was to evaluate serum concentrations of NO in patients with RA and to determine whether they correlate with clinical and laboratory parameters of RA disease activity. Twenty-seven RA patients were recruited for the study and compared with 20 healthy subjects. Serum NO concentrations were measured indirectly in terms of nitrate using colorimetric assay. Disease activity was determined by laboratory and clinical findings. Mean serum concentrations of nitrate were significantly higher than those of healthy controls (P < 0.05). Among the disease activity parameters, C-reactive protein, number of swollen and tender joints, Ritchie articular index, and disease activity scores correlated significantly with serum NO levels. Our results suggest that these levels can serve as a reliable parameter of disease activity in patients with RA. Further knowledge about the precise role of NO may lead to better understanding of the pathogenesis of RA. Furthermore, modulation of NO synthesis may represent a new approach to the treatment of inflammatory and autoimmune conditions. PMID- 11411961 TI - Keratonconjunctivitis sicca is not uncommon in children with juvenile rheumatoid arthritis. AB - Keratoconjunctivitis sicca (KCS), characterised by aqueous tear deficiency, is the most common ocular complication in adult rheumatoid arthritis (RA). In juvenile rheumatoid arthritis (JRA), however, it remains under-reported. For this prospective study, 50 children with JRA were examined clinically and underwent tests for KCS (Schirmer's I and rose bengal tests, fluorescein staining, and tear film breakup time). Six children (12%) with two or more abnormal tests were diagnosed as having definite KCS, while one child with only one abnormal test was labelled with probable KCS. Five of the six (83.3%) with definite KCS were males, and three (50%) had a pauciarticular form of the disease. Two children (33.3%) with definite KCS had no ocular symptoms, five were receiving only nonsteroidal anti-inflammatory drugs, and one was additionally on methotrexate. Keratoconjunctivitis sicca appears to be a common ocular complication and all children with JRA should be screened for it with a comprehensive battery of tests. PMID- 11411962 TI - Acute inflammatory myopathy with severe subcutaneous edema, a new variant? Report of two cases and review of the literature. AB - Acute inflammatory myopathy with severe subcutaneous edema is extremely rare and has been reported in only a handful of cases. We describe two similar patients presenting with this disorder and generalized rash. Unlike the five previously reported cases, the clinical and histologic features of our two patients are more suggestive of dermatomyositis than polymyositis. Nevertheless, scrutinizing all seven reported patients, a number of specific characteristics could be defined. All patients were adult males. Dysphagia was present in four. In six patients, acute inflammatory myopathy was idiopathic while malignancy was present in one. Two patients died despite intensive therapy, three improved on corticosteroid treatment, and two recovered spontaneously. In all patients, limb involvement with marked subcutaneous edema was present, clinically mimicking deep vein thrombosis in both our patients. The presence of severe subcutaneous edema may be a hallmark of a distinctive variant of acute inflammatory myopathy. More cases are needed to discern subtypes of this general entity and to establish guidelines for treatment and prognosis. PMID- 11411963 TI - Hughes syndrome associated with varicella infection. AB - We report a case of Hughes syndrome 1 week after the occurrence of chickenpox. The patient presented with the clinical picture of acute right iliofemoropopliteal deep vein thrombosis. IgM and IgG anticardiolipin (aCL) were both positive upon admission. The IgM aCL antibodies were still positive 6 weeks later, which suggests their role as a predisposing factor of deep vein thrombosis. PMID- 11411964 TI - Polymyositis--an unusual manifestation of chronic graft-versus-host disease. AB - Polymyositis is a rare autoimmune manifestation of chronic graft-versus-host disease (GVHD) characterized by muscle pain, weakness, and an increase in muscle related enzymes that responds well to treatment with immunosuppressive agents such as steroids and cyclosporine. We describe a case in which polymyositis was the main manifestation of chronic GVHD that occurred 12 months after allogeneic bone marrow transplantation in a patient with acute lymphocytic leukemia (ALL). The polymyositis responded well to treatment with steroids and cyclosporine, with no relapse of symptoms on tapering of the medication. PMID- 11411965 TI - Azathiopine-steroid combination therapy for pulmonary arterial aneurysms in Behcet's disease. AB - Vasculitis is the primary pathological feature of Behcet's disease and in which pulmonary arterial aneurysm formations are rare but serious complications representing poor prognosis. There is no agreement on how to treat major venous and arterial disease. The use of steroids alone or corticosteroid treatment supplemented with cyclophosphamide or azathioprine is still controversial. We present three young male patients with Behcet's disease with aneurysms in the pulmonary arteries. In two of them, cyclophosphamide was the first choice of treatment, but this was ceased due to side effects. All three patients were then treated with azathioprine (2 mg/kg per day) and methylprednisolone (1.5 mg/kg per day). All three showed clinical improvement. Regression was observed radiologically in two (one minimal and the other complete). No complication related to therapy was encountered. Steroid therapy supplemented with azathioprine may be a good choice for treatment of pulmonary arterial aneurysms in Behcet's disease. PMID- 11411966 TI - Celebrating diversity/eliminating disparity. PMID- 11411967 TI - How to "activate" medical students in the office teaching setting: giving students permission to be activate learners. PMID- 11411968 TI - Home visit. PMID- 11411969 TI - Literature and movies for medical students. PMID- 11411970 TI - The role of race in the clinical presentation. AB - What role, if any, should race play in clinical presentations? While race is widely used as a way of identifying patients, this practice has been challenged as conceptually flawed, potentially misleading, and possibly prejudicial to the patient. There are, however; important reasons for not excluding information about race. This article includes a set of guidelines for the inclusion of racial data in presentations: (1) Race is a social construct and, if used, should be recorded in the social history, not the opening sentence of the presentation. (2) Patients should self-identity their race or races. (3) Race should not be used as a proxy for genetic variation, social class, or other elements of the social history. (4) Clinicians should be mindful of the potential influence of racism in the clinical encounter. PMID- 11411971 TI - Does a reduction in family medicine clerkship time affect educational outcomes? AB - BACKGROUND AND OBJECTIVES: Little is known about the relationship between the length of a family medicine clerkship and its educational outcomes. After our family medicine clerkship time decreased from 6 weeks to 4 weeks in July 1997, we studied how this change in clerkship length affected educational outcomes. METHODS: Educational outcomes for the 2-year periods before and after the change were examined and compared whenever possible. Outcome measurements included student ratings of different aspects of the clerkship and student performance on clerkship examinations. RESULTS: Students' exposure to common clinical problems was unaffected by the change. For the 4-week clerkship, there was a slight increase in student ratings of the adequacy of number of patients seen, the opportunity to follow-up with patients, the ability to develop health promotion plans, and overall satisfaction. Because the combinations of examinations used differed each year, student performance on clerkship examinations could not be directly compared. CONCLUSIONS: Educational outcomes of the 4-week clerkship were similar to the 6-week clerkship. A few key outcomes improved. Various curricular and structural changes instituted for the 4-week clerkship contributed to the stability in outcomes. Reports from other medical schools may give additional insight into understanding this relationship. PMID- 11411972 TI - Deception by applicants to family practice residencies. AB - BACKGROUND AND OBJECTIVES: Our residency program experienced applicants who actively misrepresented qualifications, background, or abilities. The experiences of other family practice residencies were unknown. This study 1) determined what information family practice residency directors required from applicants, 2) assessed whether this information was confirmed or verified, and 3) describes the deceptive application information discovered by program directors. METHODS: A questionnaire was sent to directors of all accredited family practice residencies. Information was collected about required information, data confirmation, and the deception and misrepresentation perceived. RESULTS: Thirty two percent of the directors responded. Eight of 13 items on the Electronic Residency Application System were designated as required by more than half of responding directors. Only two of the items (licensure and certain facts in the personal statement) were confirmed by a majority of directors who required them. Deception was recognized by nearly half of respondents within the past 5 years. Most cases involved specialty choice or the personal statement of candidates and were recognized during the interview or by direct confirmation of data. CONCLUSIONS: Most directors appear to accept application information at face value. Recognition of deception about application information was reported. Misrepresentation by applicants may be a more common event than previously realized and may require more thorough verification of application credentials. PMID- 11411973 TI - EKG analysis skills of family practice residents in the United Arab Emirates: a comparison with US data. AB - BACKGROUND AND OBJECTIVES: Concern has been raised about the electrocardiogram (EKG) analysis skills of family practice residents in the United States. This study examined EKG analysis skills of family practice residents, medical students, interns, and general practitioners (GPs) in the United Arab Emirates (UAE), a different environment. METHODS: The measurement instrument was a set of 10 EKGs, used in a study of US family practice residents. Two of the EKGs were normal, and there were 14 clinical abnormalities in the remainder. RESULTS: There was no significant difference in the correct diagnosis of acute myocardial infarction between US family practice residents and UAE family practice residents, medical students, or GPs. Interns' diagnoses were significantly poorer. The mean score for correctly identifying acute myocardial infarction and both normal EKGs was not significantly different between groups: 2.50 medical students, 2.35 interns, 2.58 UAE family practice residents, 2.67 FD, and 2.55 US family practice residents. However, the US family practice resident mean score of 11.26 for all 16 clinical findings was significantly higher than any group in the UAE: 5.35 medical students, 5.87 interns, 6.08 UAE family practice residents, 5.69 family physicians. CONCLUSIONS: Difficulty in EKG interpretation transcends geographic boundaries, suggesting that new approaches to teaching these skills need to be explored. Improved EKG reading skills by family physicians are generally needed in both the United States and the UAE. PMID- 11411974 TI - Development of a residency/MPH program. AB - BACKGROUND AND OBJECTIVES: Public health complements the care physicians provide to patients. Few residencies, with the exception of preventive medicine residencies, offer trainees the opportunity to obtain a public health degree. The University of California, San Francisco-Fresno (UCSF-Fresno) Family Practice Program and California State University-Fresno (CSU-Fresno) spearheaded the creation of a combined residency/MPH program at the UCSF-Fresno Medical Education Program. METHODS: We developed a combined residency/MPH program that allows family practice residents to obtain an MPH degree during their residency training years. We describe the development process, which included initiation of the program, setting goals and objectives, identifying MPH course content and funding, and selecting applicants for entry into the program. RESULTS: The program was successfully funded, and the course content was developed. Participant selection, registration, and enrollment procedures have now been developed. Performance standards have been established, and scheduling conflicts have been addressed. The program has thus far enrolled 29 residents and faculty. Nearly one third have dropped out of the MPH component of the program, mostly because of the workload involved in simultaneous residency and MPH training. CONCLUSIONS: Other training programs can replicate a combined residency/MPH program if a strong relationship with a nearby university offering a MPH degree can be forged. The experience at UCSF-Fresno can help guide others contemplating a combined residency/MPH program. PMID- 11411975 TI - Female and underrepresented minority faculty in academic departments of family medicine: are women and minorities better off in family medicine? AB - BACKGROUND: Several studies have shown that the percentage of women represented in senior academic positions at US medical schools is lower than the percentage of men in senior positions. Similarly, the percentage of minority faculty members represented in senior academic positions is lower than that of their majority counterparts. This study assessed whether these findings were also present in departments of family medicine and identified any factors related to the institution or department that favored academic success for women and minorities. METHODS: Data regarding faculty workforce composition, including faculty rank and rank for women and underrepresented minorities, were extracted from a comprehensive survey of departments of family medicine at US allopathic medical schools. The data are based on faculty workforce in 1997 and include responses from 58 (51%) of all schools with a department of family medicine. RESULTS: Faculty in departments of family medicine were more likely to be female (41% versus 25%) and an underrepresented minority (9% versus 4%), compared with all academic medicine disciplines. However, women in full-time positions were less likely than men, and minorities were less likely than nonminorities, to be either an associate or full professor. We could find no institutional or departmental characteristics that were associated with academic success for women or minority faculty members. CONCLUSIONS: While women and underrepresented minorities are more common to the faculty workforce in family medicine, members of both of these groups are not well represented in senior faculty ranks. PMID- 11411976 TI - A telephone needs assessment for potential high utilizers. AB - BACKGROUND AND OBJECTIVES: We conducted a telephone needs assessment of patients at risk for high utilization of health services to determine the type of intervention that might be most responsive to their needs. METHODS: Family practice patients who were classified as at high risk based on self-reported health-related quality of life (HRQOL), age, and gender received a structured telephone interview to ask about their health problems, difficulties with their health care, and types of help programs that might be useful to them. RESULTS: Of 867 adult patients randomly selected as eligible for the study, 352 completed the Duke Health Profile to measure HRQOL, 122 were classified at risk for high utilization, and 104 were interviewed by telephone. Patient-reported health problems were similar to provider-reported problems. High-risk patients reported difficulties receiving help from the clinic on issues of communication and logistics (43.9%), limitations of clinic facilities and services (26.3%), and financial concerns (26.3%). Of the highest risk patients, 45.8% expressed willingness to participate in help programs conducted over the telephone. CONCLUSIONS: Many high-risk patients might benefit from participation in a telephone intervention program. A future study is needed to measure the effectiveness of such a program to improve utilization of health services. PMID- 11411977 TI - Racism in the examination room: myths, realities, and consequences. PMID- 11411978 TI - The covalent casting of one-dimensional hydrogen bonding motifs: toward oligomers and polymers of predefined topography. AB - The covalent casting of noncovalent architectures serves to define large covalent constructs that express well-defined modes of aggregation. In the case of one dimensional hydrogen-bonding motifs, covalent casting yields molecular strands that adopt a duplex mode of aggregation. The effectiveness of this design principle is illustrated through the casting of the aminotriazine hydrogen bonding motif. These studies have led to the conception of a new family of topographically defined oligomers, which, akin to DNA, self-assemble in the form of a duplex through the action of interstrand hydrogen bonds. PMID- 11411979 TI - At nonzero temperatures, stacked structures of methylated nucleic acid base pairs and microhydrated nonmethylated nucleic acid base pairs are favored over planar hydrogen-bonded structures: a molecular dynamics simulations study. AB - The dynamic structure of all ten possible nucleic acid (NA) base pairs and methylated NA base pairs hydrated by a small number of water molecules (from 1 to 16) was determined by using molecular dynamics simulations in the NVE microcanonical and NVT canonical ensembles with the Cornell force field (W. D. Cornell, P. Cieplak, C. I. Bayly, I. R. Gould, K. M. Merz, D. M. Ferguson, D. C. Spellmeyer, T. Fox, J. E. Caldwell, P. Kollman, J. Am. Chem. Soc. 1995, 117, 5179). The presence of one water molecule does not affect the structure of any hydrogen-bonded (H-bonded) nonmethylated base pair. An equal population of H bonded and stacked structures of adenine...adenine, adenine...guanine and adenine... thymine pairs is reached if as few as two water molecules are present, while obtaining equal populations of these structures in the case of adenine...cytosine, cytosine...thymine, guanine... guanine and guanine...thymine required the presence of four water molecules, and in the case of guanine...cytosine, six. A comparable population of planar, H-bonded and stacked structures for cytosine...cytosine and thymine... thymine base pairs was only obtained if at least eight water molecules hydrated a pair. Methylation of bases changed the situation dramatically and stacked structures were favoured over H bonded ones even in the absence of water molecules in most cases. Only in the case of methyl cytosine...methyl cytosine, methyl guanine...methyl guanine and methyl guanine...methyl cytosine pairs were two, two or six water molecules, respectively, needed in order to obtain a comparable population of planar, H bonded and stacked structures. We believe that these results give clear evidence that the preferred stacked structure of NA base pairs in the microhydrated environment, and also apparently in a regular solvent, is due to the hydrophilic interaction of a small number of water molecules. In the case of methylated bases, it is also due to the fact that the hydrogen atoms most suitable for the formation of H-bonds have been replaced by a methyl group. A preferred stacked structure is, thus, not due to a hydrophobic interaction between a large bulk of water molecules and the base pair, as believed. PMID- 11411980 TI - Molecular signal transduction by conformational transmission: use of tetrasubstituted perhydroanthracenes as transducers. AB - 2,3,6,7-Tetrasubstituted cis-anti-cis perhydroanthracenes have been studied as conformational transducers for molecular signal transduction. 2,2'-Bipyridine groups attached to the perhydroanthracene through ether linkages were chosen as receptor substituents, while pyrene groups were selected as effectors. A chelation-induced triple ring flip of the perhydroanthracene could be achieved by the complexation of zinc(II) ions at the bipyridine sites of ligands 13 and 15. It was found that two pyrene substituents attached to the perhydroanthracene via a linker with an E double bond and an ester group could be used to monitor the triple ring flip. In the equatorial positions, the pyrenes are sufficiently close to form an excimer in the excited state, giving a fluorescence signal at 480 nm. In the axial positions, they are far away from each other and give mainly a monomer fluorescence signal at 380 nm. Both the bipyridine receptor and the pyrene effector are present in compound 33. The conformational switching 34-->35 (the two conformers of 33) has successfully been used for a signal transduction over a signal distance of 2 nm. PMID- 11411981 TI - Predicting when precipitation-driven synthesis is feasible: application to biocatalysis. AB - Precipitation-driven synthesis offers the possibility of obtaining high reaction yields using very low volume reactors and is finding increasing applications in biocatalysis. Here, a model that allows straightforward prediction of when such a precipitation-driven reaction will be thermodynamically feasible is presented. This requires comparison of the equilibrium constant, Keq, with the saturated mass action ratio, Zsat, defined as the ratio of product solubilities to reactant solubilities. A hypothetical thermodynamic cycle that can be used to accurately predict Zsat, in water is described. The cycle involves three main processes: fusion of a solid to a supercooled liquid, ideal mixing of the liquid with octanol, and partitioning from octanol to water. To obtain the saturated mass action ratio using this cycle, only the melting points of the reactants and products, and in certain cases the pKa of ionisable groups, are required as input parameters. The model was tested on a range of enzyme-catalysed peptide syntheses from the literature and found to predict accurately when precipitation-driven reaction was possible. The methodology employed is quite general and the model is therefore expected to be applicable to a wide range of other (bio)-catalysed reactions. PMID- 11411982 TI - [Ru(HNO)('py(bu)S4')], the first HNO complex resulting from hydride addition to a NO complex ('pybuS4'2-=2,6-Bis(2-mercapto-3,5-di-tert-butylphenylthio)dime thylpyridine(2-1)). AB - Treatment of the nitrosyl complex [Ru(NO)('pybuS4')]Br (1a) with NaBH4 in CH3OH yielded [Ru(HNO)('pybuS4')](2), which could be completely characterized. The X ray structure determination of 2 confirmed the N coordination of the HNO ligand. Density functional theory calculations enabled us to assign the nu(NO) IR band of 2, which appears in KBr at 1358cm(-1) and in THF at 1378 cm(-1). The unprecedented hydride addition to nitrosyl complexes yielding HNO complexes fills a white spot on the map of chemical reactions, represents the as yet unknown counterpart to the well-established formyl complex formation from CO complexes and hydrides, and distinctly differs from the formation reaction of [Os(HNO)(CO)Cl2(PPh3)2], the only other HNO complex characterized structurally. The HNO complex 2 is oxidized stepwise by [Cp2Fe]PF6 in the presence of NEt3 and directly by Bronsted acids to give [Ru(NO)('pybuS4')]+ in 2e- oxidations. H+/D+ exchange indicates acidity of the HNO proton. PMID- 11411983 TI - Metal-modified nucleobase pairs and triplets as cytosine receptors. AB - A preorganized cationic receptor 2 for cytosine (C) is described which is composed of trans-a2PtII (a= NH3 or CH3NH2) cross-linked modules with adenine (A), guanine (G), and uracil (U) or thymine (T) model nucleobases. The functions of these three modules are as follows: i) Adenine orientates the two other bases at right angles, thus producing the L-shape of the receptor. ii) Guanine is the primary receptor. iii) Uracil or thymine act as coreceptors. Compared with the normal Watson-Crick pair between G and C, the association constant between 2 and C increases by a factor of 3 (in DMSO). As deduced from 1H NMR spectroscopy and confirmed by the X-ray crystal structure of the C adduct 4b, cytosine is fixed through five hydrogen bonds to the receptor, one of which involves the aromatic H(5) of C. A comparison of C binding is made with a structurally related linkage isomer receptor as well as the precursor molecule trans[alpha2PtAG]2+. The potential of modular, cationic receptors is illustrated. PMID- 11411984 TI - Photoinduced chain reactions of alcohols in the presence of diphenyliodonium ion pairs with cyanometallates--steady state UV/visible spectroscopic and pulse radiolysis studies. AB - Diphenyliodonium ions (Ph2I+) form donor-acceptor ion pairs with suitable cyanometallates such as [Mo(CN)8]4-, [W(CN)8]4-, [Ru(CN)6]4- and [Os(CN)6]4-. Such ion pairs are characterized by new spectroscopic transitions due to second sphere interactions between donor ([M(CN)x]4-, x=6, 8) and acceptor (Ph2I+) ions. Photochemical excitation of these ion-pair charge-transfer (IPCT) states leads to efficient electron transfer reactions that yield short-lived diphenyliodyl radicals (Ph2I*) and oxidized cyanometallates ([M(CN)x]3-). Diphenyliodyl radicals decay to iodobenzene and phenyl radicals. This very convenient source for generating phenyl radicals was applied to the photoinduced chain oxidation of primary and secondary alcohols to the corresponding aldehydes and ketones, respectively. However, unexpected side reactions led to undesired chain terminating reactions. Adduct formation of diphenyliodonium ions with alpha hydroxyalkyl radicals was verified by pulse radiolysis studies. These relatively longlived adducts give rise to chain-terminating reactions because of interactions with [M(CN)x]4- complexes that lead to oxidized cyanometallates [M(CN)x]3- upon regeneration of the starting alcohols. PMID- 11411985 TI - Enhanced intersystem crossing in donor/acceptor systems based on zinc/iron or free-base/iron porphyrins. AB - The deactivation pathways of the singlet excited state of a series of zinc or free-base donor porphyrins covalently linked by a bridge to a paramagnetic iron(III) chloride porphyrin acceptor have been studied. These donor-bridge acceptor systems all share a similar geometry (25 A donor-acceptor center-to center distance), but the bridges vary in electronic structure. In previously reported investigations of zinc/iron porphyrin systems, the fluorescence quenching of the donor has predominantly been assigned to electron transfer. However, for the porphyrin systems studied in this paper, we show that the dominant deactivation channels are enhanced intersystem crossing and singlet energy transfer. In both series, the intersystem crossing rate (S1-->T1) of the donor moiety is almost doubled in the presence of a paramagnetic high-spin metal porphyrin acceptor. The significant spectral overlap of the donor fluorescence and acceptor absorption in both series allows for efficient singlet energy transfer (Forster mechanism). Furthermore, the bridging chromophores mediate energy transfer and the enhancement is inversely dependent upon the energy gap between the donor and bridge excited states. Although Marcus theory predicts thermodynamically favorable electron transfer to occur in the systems investigated, the quenching rate constants were found to be independent of solvent polarity, and no charge-separated state could be detected, indicating very small electronic coupling for electron transfer. PMID- 11411986 TI - Rate and mechanism of the oxidative addition of phenyl iodide to Pd0 ligated by triphenylarsine: evidence for the formation of a T-shaped complex. AB - The oxidative addition of phenyl iodide to the palladium(o) generated from [Pd0(dba)2] and n equivalents of AsPh3 (the most efficient catalytic precursor in Stille reactions) proceeds from [(solv)Pd0(AsPh3)2] (solv= solvent). However, the latter is present only in trace concentrations because it is involved in an equilibrium with the major, but nonreactive, complex [Pd0(dba)(AsPh3)2]. As regards the phosphine ligands, dba has a decelerating effect on the rate of the oxidative addition by decreasing the concentration of the reactive species. Relative to PPh3, the effect of AsPh3 is to increase the rate of the oxidative addition of PhI by a factor ten in DMF and seven in THF, independent of the value of n, provided that n > or = 2. In contrast to PPh3, the addition of more than two equivalents of AsPh3 to [Pd0(dba)2] (dba= trans,trans-dibenzylideneacetone) does not affect the kinetics of the oxidative addition because of the very endergonic displacement of dba from [Pd0(dba)(AsPh3)2] to form [Pd0(AsPh3)3]. The complex trans-[PhPdI(AsPh3)2], formed in the oxidative addition, is involved in a slow equilibrium with the T-shaped complex [PhPdI(AsPh3)] after appreciable decomplexation of one AsPh3. Under catalytic conditions, that is, in the presence of a nucleophile, such as CH2=CH-SnBu3 which is able to coordinate to [Pd0(AsPh3)2], a new Pd0 complex is formed: [Pd0(eta2-CH2=CHSnBu3)(AsPh3)2]; however, this complex does not react with PhI. Consequently, CH2=CH-SnBu3 slows down the oxidative addition by decreasing the concentration of the reactive species [(solv)Pd0(AsPh3)2]. This demonstrates that a nucleophile may be not only involved in the transmetallation step, but may also interfere in the kinetics of the oxidative addition step by decreasing the concentration of reactive Pd0. PMID- 11411987 TI - Copper(II) complexes of aminocarbohydrate beta-ketoenaminic ligands: efficient catalysts in catechol oxidation. AB - Copper(II) complexes of tridentate dianionic beta-ketoenaminic ligands derived from differently functionalized amino-deoxyglucoses were synthesized and characterized with respect to their structural, spectroscopic, and catalytic properties. The (probably dimeric) complex [1,2-O-isopropylidene-6-N-(3-acetyl-2 oxobut-3-enyl)amino-6-deoxyglucofuranoso)copper(II) Cu(3a) was a highly efficient catalyst for the catechol-oxidase-like oxidation of 3,5-di-tert-butylcatechol (dtbc) into 3,5-ditert-butylquinone (dtbq) by molecular oxygen (kcat=2.63s(-1)). In contrast to this magnetically "normal" complex Cu(3a), the analogous dinuclear complex [[Cu(2a)]2], derived from the isomeric amino sugar 5-amino-5 deoxyglucofuranose, forms six-membered chelate rings with the sugar moiety and has very strong antiferromagnetic-coupled copper atoms (resulting in a diamagnetic ground state). It has a rather insignificant activity (kcat < 10( 3)s(-1)). The ligand H2 1a, derived from a (protected) 6-amino-6 deoxyglucopyranose, forms a trinuclear complex [[Cu(1a)]]2.Cu(OAc)2] in which two basic formula units are bridged by one copper acetate. This compound and the complex derived from an (isomeric) aminodeoxyglucopyranose ([Cu(4a)]: kcat approximately equals 0.03 s(-1)) show moderate activity. All complexes with a peripheral ethoxycarbonyl group instead of the acetyl substituent R2, Cu(1b) Cu(3b) and Cu(4c), are inactive. The complexes derived from 2 hydroxocyclohexylamine, Cu(5a) and Cu(5c), which were used as models of the active complex Cu(3a), have the typical "cubane-like" tetranuclear structure known from many copper complexes with derivatives of saturated 2-aminoalcohols. They are inactive with respect to the activation of dioxygen. PMID- 11411988 TI - Asymmetric reduction of ketones with catecholborane using 2,6-BODOL complexes of titanium(IV) as catalysts. AB - Reductions performed with Ti(IV) complexes of ligands based on bicyclo[2.2.2]octane diols 5 and 6 are effective catalysts in the reduction of prochiral ketones to optically active alcohols, with catecholborane as the reducing agent. Methyl ketones are favored and enantiomeric excesses (ee) of < or =98% have been achieved with acetophenone as the substrate. Several other substrates were tested, among them 2-octanone, which gave 2-octanol in 87% ee. Further details of the method were examined, for example, temperature, solvent composition, amount of molecular sieves (4 A), and catecholborane quality, as well as the sensitivity of the ligands towards acids. NMR spectroscopic methods were used to gain some insight into the complexes formed between the ligands and [Ti(OiPr)4]. A dimeric structure is proposed for the pre-catalyst. PMID- 11411989 TI - Tl-Pt(CN)5 in the solid state--A multimethod study of an unusual compound containing inorganic wires. AB - The crystal and molecular structure of a polycrystalline powder with a metal metal bond and the composition TlPt(CN)5 has been determined by combining results from X-ray powder diffraction (XRD), extended X-ray absorption fine structure (EXAFS) and vibrational spectroscopic studies. The XRD data gave the tetragonal space group P4/nmm (No. 129), with a= 7.647(3), c=8.049(3) A, Z=2, and well determined positions of the heavy metal atoms. The Pt-TI bond length in the compound is 2.627(2) A. The platinum atom coordinates four equivalent equatorial cyano ligands, with a fifth axial CN ligand and a thallium atom completing a distorted octahedral coordination geometry. The Tl-Pt(CN)5 entities are linked together in linear -NC-Pt-Tl-NC-Pt-Tl chains through the axial cyano ligand. These linear "wires" are the essential structural features and influence the properties of the compound. A three-dimensional network is formed by the four equatorial cyano ligands of the platinum atom that form bridges to the thallium atoms of neighbouring antiparallel chains. The platinum atom and the five nitrogen atoms from the bridging cyano groups form a distorted octahedron around the thallium atom. EXAFS data were recorded at the Pt and Tl L(III) edges for a more complete description of the local structure around the Pt and Tl atoms. The excessive multiple scattering was evaluated by means of the FEFF program. Raman and infrared absorption spectroscopy reveal strong coupling of the vibrational modes of the TlPt(CN)5 entities, in particular the metal-metal stretching mode, which is split into four Raman and two IR bands. Factor group theory shows that a structural unit larger than the crystallographic unit cell must be used to assign vibrational bands. Intra- and intermolecular force constants have also been calculated. The compound exhibits red luminescence at 700 +/- 3 nm in glycerol and has a corresponding excitation maximum at 240 nm. X-ray photoelectron spectra (XPS) show that the metal atoms have intermediate oxidation states, Pt3.2+ and Tl1.6-, between those in the parent Pt(II) and Tl(III) species and the decomposition products, Pt(IV) and Tl(I). The solid compound TlPt(CN)5 is stable to 520 degrees C. However in presence of water, a two-electron transfer between the metal atoms results in the cleavage of the metal-metal bond at 80 degrees C, forming a Pt(IV) pentacyanohydrate complex and a monovalent thallium ion. PMID- 11411990 TI - Synthesis of ganglioside GD3 and its comparison with bovine GD3 with regard to oligodendrocyte apoptosis mitochondrial damage. AB - 2,3-Dehydroneuraminic acid derivative 5 was transformed in five efficient steps into sialyl donor 2, which has a phenylthio group on the beta-side of the 3 position for anchimeric assistance and a diethyl phosphite residue as leaving group at the anomeric carbon. The known GM3 intermediate 10 was transformed into the 4b,4c,8c-O-unprotected acceptor 3, which was then allowed to react with 2 by using TMSOTf as catalyst and acetonitrile as solvent to afford the desired tetrasaccharide 12, which has an alpha(2-8)-linkage between two neuraminic acid residues. Removal of the phenylthio group gave intermediate 13, which was transformed into O-tetraosyl trichloroacetimidate 16 as glycosyl donor. Application of the azidosphingosine glycosylation procedure furnished GD3 (1) in high overall yield. Comparison of synthetic GD3 with bovine-brain-derived GD3 showed that there were similar effects in GD3-triggered uncoupling of mitochondrial respiration and in induction of apoptosis in oligodendrocytes. PMID- 11411991 TI - Multifaceted photoreactivity of 6-fluoro-7-aminoquinolones from the lowest excited states in aqueous media: a study by nanosecond and picosecond spectroscopic techniques. AB - Nanosecond and picosecond absorption and emission spectroscopic techniques were applied to the investigation of the reactivity from the lowest excited states of some 6-fluoro-7-piperazino-4-quinolone-3-carboxylic acids (FQs) in aqueous media at neutral pH, in the absence and presence of different sodium salts. Following the detection of various transients, we proposed a mechanism for the cleavage of the carbon-fluorine bond that proceeded through different reaction pathways, dependent on the molecular structure and the characteristics of the medium. The drug lomefloxacin (LOM), a 6,8-difluoroquinolone derivative, underwent heterolytic cleavage of the C8-F bond from the excited singlet state. With the 6 monofluoroquinolone norfloxacin (NOR) and the corresponding 1,8-naphthyridinone enoxacin (ENX), the lowest singlet state was not significantly reactive and an important deactivation channel was intersystem crossing (ISC) to the triplet manifold. The lowest triplet state underwent cleavage of the C6-F bond through a solvent mediated process possibly via a cyclohexadienyl anionic adduct. In the presence of sulfite or phosphate buffer a novel defluorination mechanism, induced by electron transfer from the inorganic anions to the FQ triplet state, was observed. The correlation between the transients observed and the final photoproducts in the different media was elucidated. PMID- 11411992 TI - Novel perylene chromophores obtained by a facile oxidative cyclodehydrogenation route. AB - New perylene chromophores, phenyl-substituted diindeno[1,2,3-cd:1',2',3' lm]perylenes 5a,b and 4,4',7,7'-tetraphenyldiacenaphtho[1,2 k:1',2',k']diindeno[1,2,3-cd:1',2',3'-me]perylenes 22a,b, have been synthesized from substituted fluoranthene derivatives 3a,b and 4a,b by means of a surprisingly simple oxidative cyclodehydrogenation reaction. The resulting chromophores, when substituted with alkyl chains at the periphery, show good solubility in organic solvents, and a full characterization of the novel red, green, and blue dyes by field-desorption mass spectrometry, UV/Vis and 1H and 13C NMR spectroscopy becomes possible. PMID- 11411993 TI - Precise adjustment of nanometric-scale diffusion layers within a redox dendrimer molecule by ultrafast cyclic voltammetry: an electrochemical nanometric microtome. AB - Performing cyclic voltammetry at scan rates into the megavolt per second range allows the exploration of the nanosecond time scale as well as the creation of nanometric diffusion layers adjacent to the electrode surface. This latter property is used here to adjust precisely the diffusion layer width within the outer shell of a fourth-generation dendrimer molecule decorated by 64 [Ru(II)(tpy)2] redox centers (tpy = terpyridine). Thus the shape of the dendrimer molecule adsorbed onto the ultramicroelectrode surface can be explored voltammetrically in a way reminiscent of an analysis with a nanometric microtome. The quantitative analysis developed here applied to the experimental voltammograms demonstrates that in agreement with previous scanning tunneling microscopy (STM) studies the adsorbed dendrimer molecules are no more spherical as they are in solution but resemble more closely hemispheres resting onto the electrode surface on their diametrical planes. The same quantitative analysis gives access to the apparent diffusion coefficient featuring electron hopping between the [Ru(II)/ Ru(III)(tpy)2] redox centers distributed on the dendrimer surface. Based on the electron hopping rate constant thus measured and on a Smoluchowski-type model developed here to take into account viscosity effects during the displacement of the [Ru(II)/Ru(III)(tpy)2] redox centers around their equilibrium positions, it is shown that the [Ru(II)/Ru(III)(tpy)2] redox centers are extremely labile in their potential wells so that they may cross-talk considerably more easily than they would do in solution at an equivalent concentration. PMID- 11411994 TI - Tropospheric formation of hydroxymethyl hydroperoxide, formic acid, H2O2, and OH from carbonyl oxide in the presence of water vapor: a theoretical study of the reaction mechanism. AB - We have carried out a theoretical investigation of the gas-phase reaction mechanism of the H2COO+ H2O reaction, which is interesting for atmospheric purposes. The B3LYP method with the 6-31G(d,p) and 6-311 + G(2d,2p) basis sets was employed for the geometry optimization of the stationary points. Additionally, single-point CCSD(T)/6-311 + G(2d,2p) energy calculations have been done for the B3LYP/6-311 + G(2d,2p) optimized structures. The reaction begins with the formation of a hydrogen-bond complex that we have calculated to be 6 kcalmol(-1) more stable than the reactants. Then, the reaction follows two different channels. The first one leads to the formation of hydroxymethyl hydroperoxide (HMHP), for which we have calculated an activation barrier of deltaGa(298) = 11.3 kcalmol(-1), while the second one gives HCO + OH + H2O, with a calculated activation barrier of deltaGa(298) = 20.9 kcalmol(-1). This process corresponds to the water-catalyzed decomposition of H2COO, and its unimolecular decomposition has been previously reported in the literature. Additionally, we have also investigated the HMHP decomposition. We have found two reaction modes that yield HCOOH+H2O; one reaction mode leads to H2CO + H2O2 and a homolytic cleavage, which produces H2COOH + OH radicals. Furthermore, we have also investigated the water-assisted HMHP decomposition, which produces a catalytic effect of about 14 kcalmol(-1) in the process that leads to H2CO + H2O2. PMID- 11411995 TI - A logical concept of structure prediction derived from supramolecular polymers of alkaline Earth metal halides formed by hydrogen bonding and complexation of the metal ion. AB - Several different dimensional polymers derived from alkaline earth metal iodides are obtained as a result of supramolecular noncovalent bonding modes of the metal ion, namely complexation and hydrogen bonding. These polymers consist of complex cations linked to the halide ions by hydrogen bonds of the water ligands coordinated to the metal. They are built up in a logical way, depending on the ratio of complexing ligands to complexing and hydrogen-bonding ligands so that their dimensionality and, to a certain extent, their structure can be predicted. PMID- 11411996 TI - Fluorescent J-type aggregates and thermotropic columnar mesophases of perylene bisimide dyes. AB - A series of perylene tetracarboxylic acid bisimides 3a-e bearing 3,4,5 tridodecyloxyphenyl substituents on the imide N atoms and zero, two, or four phenoxy-type substituents in the bay positions of the perylene core were synthesized. From investigations of their spectroscopic properties and aggregation behavior in low-polarity solvents by absorption and fluorescence optical spectroscopy, not only were these compounds found to form fluorescent J type aggregates, but also binding constants for aggregation could be derived which reflect the number and steric demand of the phenoxy substituents for bisimides 3a-d. In the pristine state, 3a-d form thermotropic hexagonal columnar mesophases which exist over a broad temperature range from below -30 degrees C to over 300 degrees C. For the tetraphenoxy-substituted compound 3e, however, a layered crystalline structure was found. This difference in behavior can be explained by the concept of microphase segregation of the aromatic cores of the molecules and the alkyl chains at the periphery. The high stability and bright fluorescence of the mesophase of several of the compounds make them promising for applications as polarizers or components in (opto)electronic devices. PMID- 11411997 TI - A combined application of reaction prediction and infrared spectra simulation for the identification of degradation products of s-triazine herbicides. AB - Substance identification in analytical chemistry is usually performed by comparing an experimental spectrum with a reference spectrum. Especially in environmental chemistry, reference spectra from databases are only available for a limited number of compounds. The combination of the reaction prediction system EROS and of infrared spectra simulation is a powerful tool for computer-assisted substance identification. First, possible degradation products of a chemical are predicted and then the infrared spectra of all these compounds are simulated. Comparison of the simulated infrared spectra with experimental spectra allows one to identify the structure of compounds. The method is demonstrated with the example of s-triazine herbicides. PMID- 11411998 TI - The 12,13-diol cyclization approach for a truly stereocontrolled total synthesis of epothilone B and the synthesis of a conformationally restrained analogue. AB - A highly convergent and stereocontrolled synthesis of epothilone B (1) has been developed. The epoxide moiety in 1 was generated by regioselective mesylation and base treatment of the 12,13-diol 30 which was formed by a chelate Cram controlled Grignard addition of 14 and methyl ketone 13. Both fragments were synthesized from the chiral carbon pool precursors (S)-citronellol and (S)-lactic acid, respectively. A highly diastereoselective aldol addition of epoxy-aldehyde 7 and the known Southern hemisphere ketone 8 delivered the full carbon skeleton, containing all the stereogenic centers of 1. Functional group manipulation, macrolactonization and removal of two protecting groups then yielded 1. The spatial closeness of the C4-beta-methyl and C6-methyl group in the crystal structure of 1 inspired us to connect them through a methylene bridge to give a cyclohexanone derivative. Thus, the Northern hemisphere aldehyde 7 was added to the enolate of the cyclohexanone 47. Further manipulations and macrolactonization delivered the conformationally restrained epothilone derivative 42. PMID- 11411999 TI - Method dependence of apparent stoichiometry in the binding of salicylate ion to human serum albumin: a comparison between equilibrium dialysis and fluorescence titration. AB - The binding of salicylate ion to human serum albumin (HSA) was studied in 100 mM potassium phosphate buffer (pH 7.4, 25 degrees C), using equilibrium dialysis and fluorescence titration methods. The protein samples tested were (a) dialyzed human plasma and (b) a commercial preparation of HSA, essentially free of globulin and fatty acids. Independent of the analytical method used, Scatchard and nonlinear regression analyses of the data pointed to a single class of high affinity salicylate binding sites. On the other hand, the binding parameters were found to be method dependent. K(d) ranged between 25 +/- 2.4 and 62 +/- 15 microM in equilibrium dialysis and between 10 +/- 1.3 and 40 +/- 3.0 microM in fluorescence titration. (The higher limits refer to plasma samples at high [HSA]). Following the same pattern, the apparent stoichiometry of binding (though independent of sample identity and concentration) was higher in equilibrium dialysis (n(app) = 3.2 +/- 0.10) than in fluorescence titration (n(app) 1.9 +/- 0.30). The difference between the two methods could be reconciled by invoking two distinct classes of binding sites (I and II), which had identical (or marginally different) K(d) values, while differing in the magnitude of the fluorescence signal (Deltaf) generated upon ligand binding (Deltaf, PL(I) = Deltaf(I); Deltaf, PL(II) = 0). Further, it was assumed that the state of occupation of class II sites affected the fluorescence efficiency of class I sites, such that Deltaf, PL(I,II) = betaDeltaf(I) (beta = interaction factor). A random binding scheme involving P, PL(I), PL(II), and PL(I,II) was formulated. The model adequately predicted the behavior of the system when monitored through the change in protein fluorescence: Taking K(d) = 25 microM and n(T) = 3, the interaction factor beta was found to be 0.62 +/- 0.10. It was concluded that the correct parameters for the binding of salicylate ion to HSA are K(d) = 25 +/- 2.4 microM and n(T) = 3.2 +/- 0.10, as indicated by equilibrium dialysis of purified HSA. Besides updating information relating to the salicylate binding potential of HSA, this study serves to illustrate a likely complication in the study of protein-ligand interactions by fluorometric methods. PMID- 11412000 TI - Femtomole peptide mapping by derivatization, high-performance liquid chromatography, and fluorescence detection. AB - A highly sensitive peptide mapping method using derivatization and fluorescence detection is described. Bovine cytochrome c was digested using a buffer compatible with the derivatization that followed. The derivatization was performed with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. The peptide mapping of the tagged digest was conducted with both HPLC and capillary LC (CLC) systems. A capillary LC-electrospray ionization mass spectrometer (MS) was set up for measuring the molecular weights of the tagged peptides. Optimization was made of the conditions used for digestion, derivatization, and mapping. MS measurements of the tagged peptides suggested that there was only one derivatization product produced from all peptides (except one) and that all the identified peptides were fully tagged. Peptide mapping of the tagged digest reviews a larger number of peptides, covering almost the entire sequence. Peptide mapping of a 20 fmol amount of tagged digest was readily performed with the CLC system. By using derivatization and fluorescence detection, the sensitivity of peptide mapping could be improved 2000 times compared to that observed with uv detection of untagged peptides. PMID- 11412001 TI - A novel reagentless sensing system for measuring glucose based on the galactose/glucose-binding protein. AB - The galactose/glucose-binding protein (GBP) is synthesized in the cytoplasm of Escherichia coli in a precursor form and exported into the periplasmic space upon cleavage of a 23-amino-acid leader sequence. GBP binds galactose and glucose in a highly specific manner. The ligand induces a hinge motion in GBP and the resultant protein conformational change constitutes the basis of the sensing system. The mglB gene, which codes for GBP, was isolated from the chromosome of E. coli using the polymerase chain reaction (PCR). Since wild-type GBP lacks cysteines in its structure, introducing this amino acid by site-directed mutagenesis ensures single-label attachment at specific sites with a sulfhydro specific fluorescent probe. Site-directed mutagenesis by overlap extension PCR was performed to prepare three different mutants to introduce a single cysteine residue at positions 148, 152, and 182. Since these residues are not involved in ligand binding and since they are located at the edge of the binding cleft, they experience a significant change in environment upon binding of galactose or glucose. The sensing system strategy is based on the fluorescence changes of the probe as the protein undergoes a structural change on binding. In this work a reagentless sensing system has been rationally designed that can detect submicromolar concentrations of glucose. The calibration plots have a linear working range of three orders of magnitude. Although the system can sense galactose as well, this epimer is not a potential interfering substance since its concentration in blood is negligible. PMID- 11412002 TI - Delay of intracellular calcium transients using a calcium chelator: application to high-throughput screening of the capsaicin receptor ion channel and G-protein coupled receptors. AB - Whole-cell functional assays are often used for high-throughput screening (HTS) of molecular targets such as ion channels and G-protein-coupled receptors. A common method for assaying the activity of these membrane proteins is to measure the change in intracellular calcium concentration upon receptor stimulation. These changes in calcium concentration are typically transient and therefore not readily adapted to high-density plate formats used in HTS instruments. We have demonstrated that an intracellular calcium chelator, BAPTA, was able to delay by 5- to 20-fold and extend for several minutes the observed calcium signals initiated by extracellular calcium influx or release of calcium from intracellular stores. As examples, we used cells expressing a calcium-permeable ion channel, vanilloid receptor type 1 (the capsaicin receptor), and two G protein-coupled receptors. These receptor-mediated increases in intracellular calcium concentration were measured by both fluorescence-based and luminescence based detection methods. The use of an intracellular calcium chelator to delay calcium signaling should have wide application since it allows the measurement of the functional activity of any cellular receptor that signals through calcium. With this procedure, calcium fluorescence and luminescence whole-cell functional assays may be performed with standard laboratory pipetting and detection systems. PMID- 11412003 TI - Effects of gum arabic on lipase interfacial binding and activity. AB - We investigated the surface behavior of gum Arabic (GA) as well as its effects on the lipolytic activity of human pancreatic lipase (HPL) and Humicola lanuginosa lipase (HLL), using emulsions of triacylglycerols (TAG) with various chain lengths. The effects of GA on the interfacial binding of HPL were also investigated. In the presence of 4 mM sodium taurodeoxycholate (NaTDC), GA (3% w/v, final concentration) had no effect on the HPL activity measured in the presence of colipase, whatever the type of TAG used. However, in the absence of bile salts or at low bile salt concentrations, GA inhibited the HPL activity when trioctanoin (TC8) and purified soybean oil (PSO) were used as substrates. At 3% (w/v, final concentration), GA strongly desorbed pure HPL from the TC8 interface and the classical anchoring effect of colipase was clearly observed. Both crude and dialyzed GA solutions were found to be highly tensioactive at the air-water as well as the oil-water interface using the drop technique. In conclusion, GA, or a putative contaminant present in GA, was found to be surface active and to have similar effects to those of bile salts on the interfacial binding and activity of HPL. PMID- 11412004 TI - Renaturation and stabilization of the telomere-binding activity of Saccharomyces Cdc13(451-693)p by L-arginine. AB - Production of recombinant proteins can be valuable in studying their biological functions. However, recombinant proteins expressed in Escherichia coli sometimes form undesirable insoluble aggregates. Solubilization and renaturation of these aggregates becomes a problem that one needs to solve. Here we used recombinant Cdc13(451-693)p as example to show the presence of l-arginine during renaturation greatly enhanced the renaturation efficiency. Cdc13p is the single-stranded telomere-binding protein of yeast Saccharomyces cerevisiae. The telomere-binding domain has been mapped within amino acids 451-693 of Cdc13p, Cdc13(451-693)p. Recombinant Cdc13(451-693)p was expressed in E. coli as insoluble protein aggregates. Purification of insoluble Cdc13(451-693)p was achieved by denaturing the protein with 6 M guanidine-HCl and followed by Ni-nitrilotriacetic acid agarose column chromatography. Renaturation of Cdc13(451-693)p to the active form was achieved by dialyzing denatured protein in the presence of l-arginine. Moreover, the presence of l-arginine was also helped in maintaining the telomere binding activity of Cdc13(451-693)p. Taking together, l-arginine might have a general application in renaturation of insoluble aggregates. PMID- 11412005 TI - An enzyme-linked immunosorbent assay for the Raf/MEK1/MAPK signaling cascade. AB - The Ras-MAPK signaling cascade transmits mitogenic stimuli from growth factor receptors and activated Ras to the cell nucleus. Inappropriate Ras activation is associated with approximately 30% of all human cancers. The kinase components of the Ras-MAPK signaling cascade are attractive targets for pharmaceutical intervention. Therefore, we have developed a high-throughput, nonradioactive ELISA method to monitor Raf and MEK1 kinase activity. In this assay system activated Raf phosphorylates and activates MEK1, which in turn phosphorylates MAPK. Antibodies that specifically detect phosphorylated MAPK (vs. nonphosphorylated MAPK) made enzyme-linked immunosorbent assay (ELISA) development possible. This assay detects inhibitors of Raf and/or MEK1 and has been used to screen large numbers of random compounds. The specific target of inhibition in the Raf/MEK1/MAPK ELISA can be subsequently identified by secondary assays which directly measure Raf phosphorylation of MEK1 or MEK1 phosphorylation of MAPK. PMID- 11412006 TI - Simultaneous detection of multiple cytokines from conditioned media and patient's sera by an antibody-based protein array system. AB - We have developed a novel technique for high-throughput simultaneous screening of multiple cytokine expression based on a protein array system. Our method has the advantage of showing the specificity of enzyme-linked immunosorbent assays, sensitivity of enhanced chemiluminescence (ECL), and high-throughput of microspot. In this system, the cytokine array membranes were created by spotting capture antibodies onto the membranes. The membranes were then incubated with biological samples such as conditioned media and patient's sera. The bound proteins were then recognized by biotin-conjugated antibodies and detected by horseradish peroxidase-conjugated streptavidin coupled with ECL. Experiments demonstrated that 24 cytokines from conditioned media and patient's sera could be simultaneously detected using this new approach. This methodology should allow us to develop many high-density protein array systems to detect a variety of proteins. To validate and quantitate the expression of key molecules in a wide range of samples, we have developed conditioned medium arrays to evaluate hundreds and even thousands of samples from individual cells and patients in a single microarray. The combinations of protein arrays and conditioned medium arrays or serum arrays will provide a powerful tool to identify the protein expression profiles and rapidly validate their expression in many types and numbers of samples. PMID- 11412007 TI - Capillary gas chromatography-mass spectrometry quantitative determination of hydroxytyrosol and tyrosol in human urine after olive oil intake. AB - Recent in vitro studies have demonstrated antioxidant properties of some virgin olive oil phenolic compounds. One of the prerequisites to extrapolate these data to an in vivo situation is the knowledge of their bioavailability in humans. In the present work we describe an analytical method which enables us to perform hydroxytyrosol and tyrosol quantitative determinations in human urine. This method was successfully used in bioavailability studies of both phenolic compounds after acute olive oil administration. Virgin olive oil was administered to healthy volunteers after a low phenolic diet. The dose administered of both phenolic compounds was estimated in reference to free forms of hydroxytyrosol and tyrosol present in virgin olive oil extracts before and after being submitted to hydrolytic conditions. These conditions mimic those occurring during digestion. Urine samples were collected before and after acute olive oil intake and analyzed by capillary gas chromatography-mass spectrometry. Hydroxytyrosol and tyrosol urinary recovery increased in response to olive oil administration, obtaining maximal values in the first 4 h. Our results further indicate that hydroxytyrosol and tyrosol are mainly excreted in conjugated form, since only 5.9 +/- 1.4% (hydroxytyrosol) and 13.8 +/- 5.4% (tyrosol) of the total amounts excreted in urine were in free form. PMID- 11412008 TI - Simultaneous measurement of anandamide and 2-arachidonoylglycerol by polymyxin B selective adsorption and subsequent high-performance liquid chromatography analysis: increase in endogenous cannabinoids in the sera of patients with endotoxic shock. AB - Anandamide (ANA) and 2-arachidonoylglycerol (2-AG), two endogenous cannabinoids, can be generated by activated macrophages and platelets, respectively, in the context of endotoxic shock, and are proposed to play a crucial role in the induction of the shock-related hypotension. Taking advantage of our recently discovered function of polymyxin B (PMB) binding to ANA and 2-AG, we developed a new method for measuring ANA and 2-AG by applying PMB-immobilized beads to selectively adsorb them in biological fluids, instead of organic solvent extraction. The eluate from beads can be directly fractionated by reverse-phase high-performance liquid chromatography (HPLC), and the fractionations corresponding to authentic ANA and 2-AG are collected and derivatized with fluorogenic reagent and subsequently quantified by HPLC with fluorometric detection. The calibration graphs of ANA and 2-AG were linear over a range of 1 to 500 pmol/ml. The limits of detection for ANA and 2-AG were 20 and 50 fmol, respectively. Intraassay precision was 2.24-4.25 and 3.47-5.44%, and interassay was 4.05-6.14 and 4.92-7.28% for ANA and 2-AG, respectively. Using this method, we first determined a 4-fold and 3-fold higher level of ANA and 2-AG, respectively, in the sera of patients with endotoxic shock than in normal serum. This finding should help in elucidating the role of the endogenous cannabinoids in the hypotension of human endotoxic shock. This method is rapid, sensitive, and reliable for simultaneously quantifying ANA and 2-AG in biological fluids, and has potential for clinical usage. PMID- 11412009 TI - Interference in yeast plasma membrane protein estimation by Percoll. PMID- 11412010 TI - Analysis of 3' and 5' ends of RNA by solid-phase S1 nuclease mapping. PMID- 11412011 TI - Mutation detection in mosaic situations: RNA mismatch assay and denaturing gradient gel electrophoresis are more sensitive than conventional cycle sequencing. PMID- 11412012 TI - Understanding of literal truth, ironic criticism, and deceptive praise following childhood head injury. AB - Children with closed head injury (CHI) have semantic-pragmatic language problems that include difficulty in understanding and producing both literal and nonliteral statements. For example, they are relatively insensitive to some of the social messages in nonstandard communication as well as to words that code distinctions among mental states. This suggests that they may have difficulty with comprehension tasks involving first- and second-order intentionality, such as those involved in understanding irony and deception. We studied how 6- to 15 year-old children, typically developing or with CHI, interpret scenarios involving literal truth, ironic criticism, and deceptive praise. Children with severe CHI had overall poorer mastery of the task. Even mild CHI impaired the ability to understand the intentionality underlying deceptive praise. CHI, especially biologically significant CHI, appears to place children at risk for failure to understand language as externalized thought. PMID- 11412013 TI - Assessing working memory and language comprehension in Alzheimer's disease. AB - Studies of language impairments in patients with Alzheimer's disease have often assumed that impairments in linguistic working memory underlie comprehension deficits. Assessment of this hypothesis has been hindered both by vagueness of key terms such as "working memory" and by limitations of available working memory tasks, in that many such tasks either seem to have little relationship to language comprehension or are too confusing or difficult for Alzheimer's patients. Four experiments investigated the usefulness of digit ordering, a new task assessing linguistic working memory and/or language processing skill, in normal adults and patients with probable Alzheimer's disease. The digit ordering task was shown to be strongly correlated with the degree of dementia in Alzheimer's patients. The task correlated with measures of language processing on which patients and normal controls performed differently. The results are interpreted as indicating that linguistic representations, linguistic processing, and linguistic working memory are intertwined, such that a deficit of one (e.g., working memory) cannot be said to "cause" a deficit in the other. The implications of this approach are explored in terms of task demands in comprehension and memory measures, and interpretation of previous results in the literature. PMID- 11412014 TI - Subcortical anarthria: a case report. AB - A 56-year-old right-handed male with a history of hypertension and diabetes presented two episodes of stroke: The first affected territory was the left anterior coroidal artery (capsular and paracapsular infarcts at the level of the genu and posterior arm of the internal capsule) and the second was the right thalamus, due to a hematoma. Following the first stroke, the patient developed severe dysarthria and after the second stroke remained anarthric. The pathophysiology of the disorder is discussed, and the role of the left and right thalamus as far as speech is concerned is reviewed. PMID- 11412015 TI - Visual field asymmetries for rhyme and semantic tasks in fluent and nonfluent bilinguals. AB - A tachistoscopic study investigated hemispheric specialization among fluent and nonfluent bilinguals for rhyme and semantic tasks in both their languages. Fluent bilinguals gave faster responses and made fewer errors in their responses to the words presented in the RVF(LH) while the opposite happened for nonfluent, which indicates greater RH participation in the first stages of a second language. Fluent bilinguals performed better not only in the second but also in their native language, suggesting superior language skills on the part of fluent bilinguals. A LVF(RH) superiority was obtained for semantic tasks, indicating RH participation in semantic judgments. Another finding was a high ratio of first born children among fluent bilinguals, potentially explained in terms of parental attitudes toward first- versus later born children. PMID- 11412016 TI - Palilalia and repetitive speech: two case studies. AB - Palilalia, a disorder of speech characterized by compulsive repetitions of utterances has been found in various neurological and psychiatric disorders. It has commonly been interpreted as a defect of motor speech. This article describes palilalia and other variants of verbal repetitive behavior, such as monosyllabic iterations and conduite d'approche. The clinical features of palilalia, its prevalence in different language tasks, and the individual patterns of verbal repetitive behavior are illustrated in two patients with a long-standing cerebrovascular disease. An attempt is made to locate the origin of different forms of verbal repetitions in a standard model of speech production (Butterworth, 1980a; Garrett, 1980; Levelt, 1989) by analysis of their morphology and correlation with impairments of lexical or phonological processes. From these observations it is suggested that palilalia results from control malfunctions at the level of the Articulator, whereas other variants of pathological verbal iterations result from an impairment of the Formulator or from malfunctions of both the Articulator and the Formulator. PMID- 11412017 TI - Conversational repair in pediatric epilepsy. AB - This study examined if children with complex partial seizures disorder (CPS) and primary generalized epilepsy with absence (PGE) were impaired in the use of self initiated repair during a conversation compared to normal children. Transcriptions of speech samples of 92 CPS, 51 PGE, and 65 normal children, ages 5-16 years, were coded for self-initiated repair according to Evans (1985). The WISC-R, a structured psychiatric interview, and seizure-related information were obtained for each child. We found impaired use of repair in both the CPS and PGE groups compared to the normal subjects. The CPS patients, particularly those with a temporal lobe focus, overused self-initiated corrections of referents and syntax compared to the PGE and normal subjects. The CPS and PGE patients with frontal lobe involvement underused fillers compared to the normal children. These findings provide additional evidence that both CPS and PGE impact the ongoing development of children's communication skills. PMID- 11412018 TI - Gender and the integration of acoustic dimensions of prosody: implications for clinical studies. AB - This study was conducted to detect the existence of a relationship between spectral and temporal prosodic cues and to examine gender differences in any such relationship. The rationale for the investigation was to gain a greater understanding of normal prosody and the requirements for control groups in clinical studies of prosody. Ten male and 10 female speakers with no known speech or neurological deficits participated in the study. They performed a reading task which involved delivering 10 sentences first with a declarative and then repeated with an interrogative intonation (20 sentences per speaker). Intrasubject and intersubject analyses of the speech data revealed a dependence of pitch on duration that differed between male and female speakers. Significant differences between the genders were also found in speech rate, pitch range, and pitch slope. The findings suggest that an integrated treatment of acoustic cues may provide a more invariant feature of normal prosody against which clinical groups may be compared. The data also imply that in clinical studies of the production of prosody gender should be carefully controlled. PMID- 11412019 TI - Acquisition of lexical morphology in simple partial epilepsy. AB - In the study of language acquisition following early brain damage, results have been divergent. On one hand, some studies claim that language eventually resumes to normal, whereas, on the other hand, studies show lasting deficits throughout development. Discrepancies in the results could arise from different etiologies and tests used. This study attempts to determine the extent to which the development of verb production is affected in later development in children who had simple partial epilepsy (SPE). Measures of diversity and fluency of three verb types, namely main verbs, auxiliary and copula verbs, and nonfinite verbs were used on three children diagnosed as SPE and compared to control groups. Our main results show a limited production of auxiliary verbs. Further analysis of their productions suggest a telegraphic style of speech, as reflected by a superior production of nonfinite verb type compared with normal children. These findings are interpreted as reflecting long-lasting consequences of early brain damage with respect to language development. PMID- 11412020 TI - Social validity of changes in informativeness and efficiency of aphasic discourse following linguistic specific treatment (LST). AB - This study presents the results of an analysis of pragmatic aspects of language samples obtained from five agrammatic aphasic individuals prior to and following Linguistic Specific Treatment (LST). Clinically and statistically significant positive changes in informativeness and efficiency were documented in posttreatment samples. Analysis of communicative competence (i.e., naive listeners' ratings of pre- and posttreatment audiotaped samples), was undertaken to examine the social validity of quantified changes. Mean ratings across listeners were computed to evaluate their subjective perceptions of general communicative constructs. Results indicated that objectively measured changes in pragmatic aspects were perceptible to naive listeners, however, to varying degrees across participants and constructs. PMID- 11412023 TI - Induction of cholinergic function in cultured sympathetic neurons by periosteal cells: cellular mechanisms. AB - Periosteum, the connective tissue surrounding bone, alters the transmitter properties of its sympathetic innervation during development in vivo and after transplantation. Initial noradrenergic properties are downregulated and the innervation acquires cholinergic and peptidergic properties. To elucidate the cellular mechanisms responsible, sympathetic neurons were cultured with primary periosteal cells or osteoblast cell lines. Both primary cells and an immature osteoblast cell line, MC3T3-E1, induced choline acetyltransferase (ChAT) activity. In contrast, lines representing marrow stromal cells or mature osteoblasts did not increase ChAT. Growth of periosteal cells with sympathetic neurons in transwell cultures that prevent direct contact between the neurons and periosteal cells or addition of periosteal cell-conditioned medium to neuron cultures induced ChAT, indicating that periosteal cells release a soluble cholinergic inducing factor. Antibodies against LIFRbeta, a receptor subunit shared by neuropoietic cytokines, prevented ChAT induction in periosteal cell/neuron cocultures, suggesting that a member of this family is responsible. ChAT activity was increased in neurons grown with periosteal cells or conditioned medium from mice lacking either leukemia inhibitory factor (LIF) or LIF and ciliary neurotrophic factor (CNTF). These results provide evidence that periosteal cells influence sympathetic neuron phenotype by releasing a soluble cholinergic factor that is neither LIF nor CNTF but signals via LIFRbeta. PMID- 11412024 TI - Complementary functions of Otx2 and Cripto in initial patterning of mouse epiblast. AB - The development of the mammalian antero-posterior (A-P) axis is proposed to be established by distinct anterior and posterior signaling centers, anterior visceral endoderm and primitive streak, respectively. Knock-out studies in mice have shown that Otx2 and Cripto have crucial roles in the generation and/or functions of these anterior and posterior centers, respectively. In both Otx2 and Cripto single mutants, the initial formation of the A-P axis takes place in a proximal-distal (P-D) orientation, but subsequent axis rotation fails to occur. To examine the developmental consequences of the lack of these two genes, we have analyzed the Otx2(-/-);Cripto(-/-) double homozygous mutant phenotype. In the double mutants, the expression of the A-P axis markers Cer-l, Lim1, and Wnt3 was not induced, while expression of Fgf8 and T was expanded throughout the epiblast, indicating that the double mutants could not form the A-P axis even in its initial P-D orientation. In addition, the double mutants displayed defects in differentiation of the visceral endoderm overlying the epiblast, as well as in the extraembryonic ectoderm. Furthermore, differentiation of neuroectoderm was accelerated as judged by the reduction of Oct4 expression and emergence of Sox1 and Gbx2 expression in the double mutant epiblast. The resulting ectoderm only displayed characteristics of anterior hindbrain, implicating it as a ground state in the mammalian body plan. Our results indicate that complementary functions of Otx2 and Cripto are essential for initial patterning of the A-P axis in the mouse embryo. PMID- 11412025 TI - Activated armadillo/beta-catenin does not play a general role in cell migration and process extension in Drosophila. AB - Human beta-catenin and its fly homolog Armadillo are best known for their roles in cadherin-based cell-cell adhesion and in transduction of Wingless/Wnt signals. It has been hypothesized that beta-catenin may also regulate cell migration and cell shape changes, possibly by regulating the microtubule cytoskeleton via interactions with APC. This hypothesis was based on experiments in which a hyperstable mutant form of beta-catenin was expressed in MDCK cells, where it altered their migratory properties and their ability to send out long cellular processes. We tested the generality of this hypothesis in vivo in Drosophila. We utilized three model systems in which cell migration and/or process extension are known to play key roles during development: the migration of the border cells during oogenesis, the extension of axons in the nervous system, and the migration and cell process extension of tracheal cells. In all cases, cells expressing activated Armadillo were able to migrate and extend cell processes essentially normally. The one alteration from normal involved an apparent cell fate change in certain tracheal cells. These results suggest that only certain cells are affected by activation of Armadillo/beta-catenin, and that Armadillo/beta-catenin does not play a general role in inhibiting cell migration or process extension. PMID- 11412026 TI - Developmental change in the steroid hormone signal for cell-autonomous, segment specific programmed cell death of a motoneuron. AB - During metamorphosis of the hawkmoth, Manduca sexta, accessory planta retractor (APR) motoneurons undergo a segment-specific pattern of programmed cell death (PCD): e.g., APRs from abdominal segment six [APR(6)s] die at pupation in direct response to the prepupal rise in 20-hydroxyecdysone (20E), whereas APR(4)s survive through the pupal stage and die at eclosion (adult emergence). The hypothesis that the death of APR(4)s is triggered by the decline in 20E at eclosion was supported by findings that injection of 20E into developing pupae to delay the fall in 20E delayed APR(4) death. Furthermore, abdomen isolation to advance the fall in 20E caused precocious APR(4) death. In other experiments, APR(4)s were placed in primary cell culture 4 days before eclosion in medium containing 1 microg/ml 20E. A switch to hormone-free medium induced PCD in a significant proportion of APR(4)s, compared to APR(4)s that remained in 20E. Process fragmentation was a reliable early indicator of PCD. These results show that a decline in 20E triggers cell-autonomous PCD of APR(4)s, in contrast to the rise in 20E that triggers cell-autonomous PCD of APR(6)s. Thus, the PCD of homologous motoneurons in different body segments at different developmental times is triggered by different steroid hormone signals. PMID- 11412027 TI - Tbx1, a DiGeorge syndrome candidate gene, is regulated by sonic hedgehog during pharyngeal arch development. AB - Appropriate interactions between the epithelium and adjacent neural crest-derived mesenchyme are necessary for normal pharyngeal arch development. Disruption of pharyngeal arch development in humans underlies many of the craniofacial defects observed in the 22q11.2 deletion syndrome (del22q11), but the genes responsible remain unknown. Tbx1 is a T-box transcription factor that lies in the 22q11.2 locus. Tbx1 transcripts were found to be localized to the pharyngeal endoderm and the mesodermal core of the pharyngeal arches, but were not present in the neural crest-derived mesenchyme of the pharyngeal arches. Sonic hedgehog (Shh) is also expressed in the pharyngeal arches and is necessary for normal craniofacial development. We found that Tbx1 expression was dependent upon Shh signaling in mouse embryos, consistent with their overlapping expression in the pharyngeal arches. Furthermore, Shh was sufficient to induce Tbx1 expression when misexpressed in selected regions of chick embryos. These studies reveal a Shh mediated pathway that regulates Tbx1 during pharyngeal arch development. PMID- 11412028 TI - A novel mode of asymmetric division identifies the fly neuroglioblast 6-4T. AB - Asymmetric cell divisions and segregation of fate determinants are crucial events in the generation of cell diversity. Fly neuroblasts, the precursors that self reproduce and generate neurons, represent a clear example of asymmetrically dividing cells. Less is known about how neurons and glial cells are generated by multipotent precursors. Flies provide the ideal model system to study this process. Indeed, neuroglioblasts (NGBs) can be specifically identified and have been shown to require the glide/gcm fate determinant to produce glial cells, which otherwise would become neurons. Here, we follow the division of a specific NGB (NGB6-4T), which produces a neuroblast (NB) and a glioblast (GB). We show that, to generate the glioblast, glide/gcm RNA becomes progressively unequally distributed during NGB division and preferentially segregates. Subsequently, a GB specific factor is required to maintain glide/gcm expression. Both processes are necessary for gliogenesis, showing that the glial vs. neuronal fate choice is a two-step process. This feature, together with glide/gcm subcellular RNA distribution and the behavior of the NGB mitotic apparatus identify a novel type of division generating cell diversity. PMID- 11412029 TI - Function of Neurolin (DM-GRASP/SC-1) in guidance of motor axons during zebrafish development. AB - Neurolin (zf DM-GRASP), a transmembrane protein with five extracellular immunoglobulin domains, is expressed by secondary but not primary motoneurons during zebrafish development. The spatiotemporally restricted expression pattern suggests that Neurolin plays a role in motor axon growth and guidance. To test this hypothesis, we injected zebrafish embryos with function-blocking Neurolin antibodies. In injected embryos, secondary motor axons form a broadened bundle along the common path and ectopic branches leave the common path at right angles. Moreover, the formation of the ventral and the rostral projection of secondary motor axons is inhibited during the second day of development. Pathfinding errors, resulting in secondary motor axons growing through ectopic regions of the somites, occur along the common path and in the dorsal and rostral projection. Our data are compatible with the view that Neurolin is involved in the recognition of guidance cues and acts as a receptor on secondary motor axons. Consistent with this idea is the binding pattern of a soluble Neurolin-Fc construct showing that putative ligands are distributed along the common path, the ventral projection, and in the area where the rostral projection develops. PMID- 11412030 TI - Misexpression of noggin leads to septal defects in the outflow tract of the chick heart. AB - BMP-2 and BMP-4 are known to be involved in the early events which specify the cardiac lineage. Their later patterns of expression in the developing mouse and chick heart, in the myocardium overlying the atrioventricular canal (AV) and outflow tract (OFT) cushions, also suggest that they may play a role in valvoseptal development. In this study, we have used a recombinant retrovirus expressing noggin to inhibit the function of BMP-2/4 in the developing chick heart. This procedure resulted in abnormal development of the OFT and the ventricular septum. A spectrum of abnormalities was seen ranging from common arterial trunk to double outlet right ventricle. In hearts infected with noggin virus, where the neural crest cells have been labelled, the results show that BMP 2/4 function is required for the migration of neural crest cells into the developing OFT to form the aortopulmonary septum. Prior to septation, misexpression of noggin also leads to a decrease in the number of proliferating mesenchymal cells within the proximal cushions of the outflow tract. These results suggest that BMP-2/4 function may mediate several key events during cardiac development. PMID- 11412031 TI - Role of thyroid hormone receptors in timing oligodendrocyte differentiation. AB - The timing of oligodendrocyte differentiation is thought to depend on both intracellular mechanisms and extracellular signals. Thyroid hormone (TH) helps control this timing both in vitro and in vivo, but it is still uncertain how it does so. TH acts through nuclear receptors that are encoded by two genes, TRalpha and TRbeta. Previous studies suggested that TRbeta receptors may mediate the effect of TH on oligodendrocyte precursor cells (OPCs). Consistent with this possibility, we show here that overexpression of TRbeta1 promotes precocious oligodendrocyte differentiation, whereas expression of two dominant-negative forms of TRbeta1 greatly delays differentiation. Surprisingly, however, we find that postnatal TRbeta-/- mice have a normal number of oligodendrocytes in their optic nerves and that TRbeta-/- OPCs stop dividing and differentiate normally in response to TH in vitro. Moreover, we find that OPCs do not express TRbeta1 or TRbeta2 mRNAs, whereas they do express TRalpha1 and TRalpha2 mRNAs. These findings suggest that alpha receptors mediate the effect of TH on the timing of oligodendrocyte differentiation. We also show that TRalpha2 mRNA, which encodes a dominant-negative form of TRalpha, decreases as OPCs proliferate in vitro and in vivo. This decrease may help control when oligodendrocyte precursors differentiate. PMID- 11412032 TI - Excess lunatic fringe causes cranial neural crest over-proliferation. AB - Lunatic fringe is a vertebrate homologue of Drosophila fringe, which plays an important role in modulating Notch signaling. This study examines the distribution of chick lunatic fringe at sites of neural crest formation and explores its possible function by ectopic expression. Shortly after neural tube closure, lunatic fringe is expressed in most of the neural tube, with the exception of the dorsal midline containing presumptive neural crest. Thus, there is a fringe/non-fringe border at the site of neural crest production. Expression of excess lunatic fringe in the cranial neural tube and neural crest by retrovirally mediated gene transfer resulted in a significant increase ( approximately 60%) in the percentage of cranial neural crest cells 1 day after infection. This effect was mediated by an increase in cell division as assayed by BrdU incorporation. Infected embryos had an up-regulation of Delta-1 in the dorsal neural tube and redistribution of Notch-1 to the lumen of the neural tube, confirming that excess fringe modulates Notch signaling. These findings point to a novel role for lunatic fringe in regulating cell division and/or production of neural crest cells by the neural tube. PMID- 11412033 TI - The Drosophila melanogaster X-linked mfs(1)6E locus is required for production of normal seminal fluid by the male accessory glands. AB - The Drosophila male accessory glands (paragonias) are two male-specific organs that produce seminal fluid, a secretion involved in sperm storage and subsequent sperm utilization by the female. This paper reports the first X-linked locus, male-female-sterile in region 6E [mfs(1)6E], required for the production of normal seminal fluid. Mutant males produce motile spermatozoa, which are transferred to females during mating, but which are not stored. Sterility of these males is mainly due to severe affected transfer of seminal fluid to females during mating. In addition, the mutant seminal fluid seems defective in triggering the behavioral (reduced receptivity to further mating) and physiological (increased egg-laying) changes normally observed in mated females. Mutant male accessory glands show notable abnormalities, connected with glandular secretion as well as qualitative and quantitative differences in their protein content. PMID- 11412034 TI - Ultraviolet irradiation- and dimethyl sulfoxide-induced telomerase activity in ovarian epithelial cell lines. AB - Information about telomerase regulation is incomplete, especially since various studies suggest complexity in telomerase regulation. Given the important association between telomerase and cancer, it is imperative to design and develop a model system in which telomerase activity can be regulated and studied. We employed ultraviolet (UV) radiation or dimethyl sulfoxide (DMSO) to transiently induce telomerase activity in a telomerase-positive cell line and, most importantly, in a telomerase-negative cell line. UV- or DMSO-induced telomerase activity was associated with increased hTRT, but not hTR, mRNA transcription in the telomerase-negative cells. However, no changes in hTRT or hTR mRNA transcription were noted with UV- or DMSO-induced telomerase activity in the telomerase-positive cells. Inhibition of protein synthesis or the phosphotidyl inositol 3-kinase (PI3K) pathway suppressed telomerase induction and/or activity in all cell lines examined, suggesting telomerase activity was dependent on protein synthesis and PI3K-mediated phosphorylation. Furthermore, enhanced telomerase activity was limited to UV and DMSO, since a variety of chemotherapeutic agents failed to induce telomerase activity. Therefore, our data provide a useful culture model system to study telomerase regulation in telomerase-negative and -positive cell lines and from which to obtain information about telomerase as a target for cancer intervention. PMID- 11412035 TI - EGF-dependent translocation of green fluorescent protein-tagged PLC-gamma1 to the plasma membrane and endosomes. AB - Growth factor-dependent translocation of phospholipase C-gamma1 (PLC-gamma1) was investigated using a green fluorescent protein-tagged PLC-gamma1 (PLC-gamma1-GFP) expressed in human epidermoid carcinoma A-431 cells. In the absence of growth factors, PLC-gamma1-GFP was present throughout the cytoplasm of A-431 cells. Treatment of the cells with epidermal growth factor (EGF) produced a very rapid redistribution of PLC-gamma1-GFP to the plasma membrane in a nonuniform manner. This translocation to the plasma membrane was insensitive to an inhibitor of phosphatidylinositol 3-kinase and was independent of cell adhesion. However, the translocation was disrupted by an agent which depolymerizes the actin cytoskeleton. At later times following the addition of EGF, PLC-gamma1-GFP appeared associated with intracellular vesicles. Stimulation of A-431 cells by Texas red-conjugated EGF for more than 10 min resulted in punctate intracellular PLC-gamma1-GFP distribution that colocalized with Texas red-conjugated EGF. This suggests that PLC-gamma1 is translocated to endosomes after EGF treatment, probably by associating with the internalized and autophosphorylated EGF receptor. Fractionation studies demonstrated that the EGF-induced plasma membrane localized PLC-gamma1 is concentrated in caveolae microdomains. Disruption of caveolae with methyl-beta-cyclodextrin resulted in the ablation of EGF-induced, but not bradykinin-induced, mobilization of intracellular Ca(2+). This treatment, however, only partially decreased PLC-gamma1 membrane translocation. PMID- 11412036 TI - Agonist-dependent immobilization of chimeric bombesin/GRP receptors: dependence on c-Src activity and dissociation from internalization. AB - G-protein-coupled receptors (GPCRs) are membrane proteins that exhibit a decreased mobile fraction compared to a freely mobile plasma membrane protein. Recently, interest has focused on proteins other than heterotrimeric G-proteins that interact with GPCRs as scaffolding structures that affect receptor signal transduction. In order to investigate the physical state of receptors before and after agonist, we used fluorescence recovery after photobleaching of the bombesin/gastrin-releasing peptide (GRP) receptor fused to the intrinsically fluorescent green fluorescent protein (GFP-GRP receptor) expressed in KNRK cells to measure both the fraction of mobile receptors and their diffusion rate before and after agonist stimulation. In live cells at 37 degrees C, addition of GRP (100 nM) caused a rapid decrease in GFP-GRP receptor mobile fraction from 0.8 +/- 0.1 to 0.49 +/- 0.05, which was independent of endocytosis. Concurrently, the remaining mobile GFP-GRPreceptors showed an increase in the diffusion rate with the half-time of fluorescent recovery, tau(1/2) = 46 +/- 7 s for untreated cells, decreasing to tau(1/2) = 30 +/- 6 s for cells treated with GRP. Prior treatment with the Src-specific inhibitor PP-2 (10 microM) blocked GFP-GRP receptor immobilization while treatment with the inactive analog PP-3 (10 microM) did not affect receptor immobilization. These data suggest that agonist-bound GPCR have increased plasma membrane diffusion rates but an increased affinity for immobilization into a multiprotein complex that is mediated by Src activity. PMID- 11412037 TI - Membrane trafficking machinery components associated with the mammalian acrosome during spermiogenesis. AB - Active trafficking from the Golgi apparatus is involved in acrosome formation, both by delivering acrosomal contents to the nascent secretory vesicle and by controlling organelle growth and shaping. During murine spermiogenesis, Golgi antigens (giantin, beta-COP, golgin 97, mannosidase II) are detected in the acrosome until the late cap-phase spermatids, but are not found in testicular spermatozoa (maturation-phase spermatids). This suggests that Golgi-acrosome flow may be relatively unselective, with Golgi residents retrieved before spermiation is complete. Treatment of spermatogenic cells with brefeldin A, a drug that causes the Golgi apparatus to collapse into the endoplasmic reticulum, disrupted the Golgi in both pachytene spermatocytes and round spermatids. However, this treatment did not affect the acrosomal granule, and some beta-COP labeling on the acrosome of elongating spermatids was maintained. Additionally, N-ethylmaleimide sensitive factor, soluble NSF attachment proteins, and homologues of the t-SNARE syntaxin and of the v-SNARE VAMP/synaptobrevin, as well as members of the rab family of small GTPases, are associated with the acrosome (but not the acrosomal granule) in round and elongated spermatids. This suggests that rab proteins and the SNARE machinery for membrane recognition/docking/fusion may be involved in trafficking during mammalian acrosome biogenesis. PMID- 11412038 TI - Protein targeting to the plasma membrane of adult skeletal muscle fiber: an organized mosaic of functional domains. AB - The plasma membrane of differentiated skeletal muscle fibers comprises the sarcolemma, the transverse (T) tubule network, and the neuromuscular and muscle tendon junctions. We analyzed the organization of these domains in relation to defined surface markers, beta-dystroglycan, dystrophin, and caveolin-3. These markers were shown to exhibit highly organized arrays along the length of the fiber. Caveolin-3 and beta-dystroglycan/dystrophin showed distinct, but to some extent overlapping, labeling patterns and both markers left transverse tubule openings clear. This labeling pattern revealed microdomains over the entire plasma membrane with the exception of the neuromuscular and muscle-tendon junctions which formed distinct demarcated macrodomains. Our results suggest that the entire plasma membrane of mature muscle comprises a mosaic of T tubule domains together with sareolemmal caveolae and beta-dystroglycan domains. The domains identified with these markers were examined with respect to targeting of viral proteins and other expresseddomain-specific markers. We found that each marker protein was targeted to distinct microdomains. The macrodomains were intensely labeled with all our markers. Replacing the cytoplasmic tail of the vesicular stomatitis virus glycoprotein with that of CD4 resulted in retargeting from one domain to another. The domain-specific protein distribution at the muscle cell surface may be generated by targeting pathways requiring specific sorting information but this trafficking is different from the conventional apical-basolateral division. PMID- 11412039 TI - Effect of PPAR activators on cytokine-stimulated cyclooxygenase-2 expression in human colorectal carcinoma cells. AB - Cyclooxygenase-2 (COX-2) expression is up-regulated in colorectal cancer tissue. Peroxisome proliferator-activated receptors (PPARs) are expressed in human colorectal tissue and activation of PPARs can alter COX-2 expression. In macrophages, activation of PPARs down-regulates COX-2 expression. We examined the effect of PPARalpha and PPARgamma ligands on untreated and TNF-alpha-induced COX 2 expression in the human colorectal epithelial cell line HT-29. The expression of PPARalpha and PPARgamma was confirmed in these cells. TNF-alpha, an inflammatory cytokine, increased COX-2 expression via the NFkappaB pathway. In the absence of TNF-alpha, WY14643 (PPARalpha activator) caused an increase, while BRL49653 (PPARgamma activator) did not alter COX-2 expression. When HT-29 cells were incubated with TNF-alpha and WY14643, a further increase in COX-2 expression was detected. Incubation with TNF-alpha and BRL49653 caused an additional twofold increase in COX-2 expression. Our results suggest that both PPARalpha signaling and TNF-alpha signaling increase COX-2 expression by independent pathways, while PPARgamma stimulates COX-2 expression by up-regulation of the TNF-alpha pathway. PMID- 11412040 TI - An essential role for Src kinase in ErbB receptor signaling through the MAPK pathway. AB - ErbB receptor tyrosine kinases are activated by multiple ligands such as epidermal growth factor (EGF) and neuregulins (NRGs), leading to stimulation of intracellular signaling pathways, including the mitogen-activated protein kinase (MAPK) cascade. We show here that Src kinase is essential for rapid EGF- and NRG induced MAPK activation when the breast carcinoma cell lines T47D and SKBR3 are stimulated with low concentrations of ligand. In the presence of the pharmacological inhibitor CGP77675, which specifically blocks the activity of Src family kinases, ligand-induced MAPK activation was almost completely blocked at 5 min. Although this block was only transient, inactivation of Src suppressed ligand-induced transcription from a MAPK-responsive promoter. At the molecular level, the initial inhibition of MAPK by Src inactivation correlated with impaired ligand-induced Shc phosphorylation. Surprisingly, Src inhibition affected neither association of Shc with ErbB receptors nor phosphorylation of receptor-bound Shc. Thus, ErbB signaling requires the engagement of a novel Src dependent route to MAPK, to trigger its rapid activation and subsequent efficient stimulation of transcription. PMID- 11412041 TI - Oxygen-mediated regulation of gelatinase and tissue inhibitor of metalloproteinases-1 expression by invasive cells. AB - The relative expression of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) is an important determinant in trophoblast invasion of the uterus and tumor invasion and metastasis. Our previous studies have shown that low oxygen levels increase the in vitro invasiveness of trophoblast and tumor cells. The present study examined whether changes in oxygen levels affect TIMP and MMP expression by cultured trophoblast and breast cancer cells. Reverse zymographic analysis demonstrated reduced TIMP-1 protein secretion by HTR-8/SVneo trophoblast cells as well as MDA-MB-231 and MCF-7 breast carcinoma cells cultured in 1% vs 20% oxygen for 24 h. While gelatin zymography revealed no changes in the levels of MMP-9 secreted by HTR-8/SVneo trophoblasts cultured under various oxygen concentrations for 24 h, human MDA-MB-231 breast carcinoma cells displayed increased MMP-9 secretion and human MCF-7 breast cancer cells exhibited reduced secretion of this enzyme when cultured under similar conditions. In contrast, MMP 2 levels remained unchanged in all cultures incubated under similar conditions. Western blot analysis of MMP-9 protein in cell extracts confirmed the results of zymography. To assess the contribution of enhanced MMP activity to hypoxia induced invasion, the effect of an MMP inhibitor (llomastat) on the ability of MDA-MB-231 cells to penetrate reconstituted extracellular matrix (Matrigel) was examined. Results showed that MMP inhibition significantly decreased the hypoxic upregulation of invasion by these cells. These findings indicate that the increased cellular invasiveness observed under reduced oxygen conditions may be due in part to a shift in the balance between MMPs and their inhibitors favoring increased MMP activity. PMID- 11412042 TI - beta-Lapachone-induced apoptosis in human prostate cancer cells: involvement of NQO1/xip3. AB - beta-Lapachone (beta-lap) induces apoptosis in various cancer cells, and its intracellular target has recently been elucidated in breast cancer cells. Here we show that NAD(P)H:quinone oxidoreductase (NQO1/xip3) expression in human prostate cancer cells is a key determinant for apoptosis and lethality after beta-lap exposures. beta-Lap-treated, NQO1-deficient LNCaP cells were significantly more resistant to apoptosis than NQO1-expressing DU-145 or PC-3 cells after drug exposures. Formation of an atypical 60-kDa PARP cleavage fragment in DU-145 or PC 3 cells was observed after 10 microM beta-lap treatment and correlated with apoptosis. In contrast, LNCaP cells required 25 microM beta-lap to induce similar responses. Atypical PARP cleavage in beta-lap-treated cells was not affected by 100 microM zVAD-fmk; however, coadministration of dicoumarol, a specific inhibitor of NQO1, reduced beta-lap-mediated cytotoxicity, apoptosis, and atypical PARP cleavage in NQO1-expressing cells. Dicoumarol did not affect the more beta-lap-resistant LNCaP cells. Stable transfection of LNCaP cells with NQO1 increased their sensitivity to beta-lap, enhancing apoptosis compared to parental LNCaP cells or vector-alone transfectants. Dicoumarol increased survival of beta lap-treated NQO1-expressing LNCaP transfectants. NQO1 activity, therefore, is a key determinant of beta-lap-mediated apoptosis and cytotoxicity in prostate cancer cells. PMID- 11412043 TI - Mechanical stretch induces activation of skeletal muscle satellite cells in vitro. AB - Cultured quiescent satellite cells were subjected to mechanical stretch in a FlexerCell System. In response to stretch, satellite cells entered the cell cycle earlier than if they were under control conditions. Only a brief period of stretch, as short as 2 h, was necessary to stimulate activation. Additionally, conditioned medium from stretched cells could activate unstretched satellite cells. The presence of HGF on c-met-positive myogenic cells was detected by immunofluorescence at 12 h in culture, and immunoblots demonstrated that HGF was released by stretched satellite cells into medium. Also, stretch activation could be abolished by the addition of anti-HGF antibodies to stretched cultures, and activity in conditioned medium from stretched cells could be neutralized by anti HGF antibodies. In addition, stretch appeared to cause release of preexisting HGF from the extracellular matrix. These experiments suggest that HGF may be involved in linking mechanical perturbation of muscle to satellite cell activation. PMID- 11412044 TI - Diverse roles of conserved asparagine-linked glycan sites on tyrosinase family glycoproteins. AB - The tyrosinase family of genes has been conserved throughout vertebrate evolution. The role of conserved N-glycan sites in sorting, stability, and activity of tyrosinase family proteins was investigated using two family members from two different species, mouse gp75/tyrosinase-related protein (TRP)-1/Tyrp1 and human tyrosinase. Potential N-linked glycosylation sites on the lumenal domains of mouse gp75/TRP-1/Tyrp1 and human tyrosinase were eliminated by site directed mutagenesis (Asn to Gln substitutions). Our results show that selected conserved N-glycan sites on tyrosinase family members are crucial for stability in the secretory pathway and endocytic compartment and for enzymatic activity. Different glycan sites on the same tyrosinase family polypeptide can perform distinct functions, and conserved sites on tyrosinase family paralogues can perform different functions. PMID- 11412045 TI - hVPS41 is expressed in multiple isoforms and can associate with vesicles through a RING-H2 finger motif. AB - Vps41p, the protein encoded by the yeast gene VPS41, has been shown to mediate formation of AP-3 transport vesicles from the Golgi apparatus and to facilitate the docking and fusion of lysosomal vesicles. Although both of these activities involve transient association with membrane structures, the mechanisms that mediate those interactions have not been determined. Orthologues of VPS41 have been identified in humans, Drosophila, tomato, and Arabidopsis; the degree of sequence similarity among these genes suggests a highly conserved function. Here we provide evidence that hVps41, the human homologue of Vps41p, is expressed in two isoforms that differ in that one contains a C-terminal RING-H2 sequence motif. Transient expression analysis suggests that this RING-H2 domain is responsible for membrane association. This observation was further supported by the cytosolic localization of site-specific mutants. A truncated construct containing only the hVps41 RING-H2 domain was found to associate with a class of intracellular vesicles that originated from the Golgi and showed partial coincidence with the delta subunit of the adaptor protein complex-3. Together with information from the homologous yeast system, these results suggest that hVps41 may also be involved in the formation and fusion of transport vesicles from the Golgi. PMID- 11412046 TI - Characterization of a dominant inhibitory E47 protein that suppresses C2C12 myogenesis. AB - Skeletal muscle formation is controlled through the coordinated actions of the muscle regulatory factors (MRFs). The activities of these basic helix-loop-helix proteins is mediated in part through heterodimer formation with a family of ubiquitous bHLH proteins, referred to as E-proteins. The primary E-protein in skeletal muscle is the E2A splice variant, E47. To further address the role of E47 during skeletal myogenesis, we created a chimeric E47 repressor protein by replacing the transcriptional activation domain with the Drosophila Engrailed transcriptional repressor domain. The dominant inhibitory E-protein (EnDeltaE47) formed homodimers capable of binding DNA and abolished E47-directed gene transcription. Stable expression of EnDeltaE47 in mouse C2C12 myoblasts effectively blocked the cells' ability to differentiate into mature myofibers. Closer examination of the molecular basis for the inhibition of myogenesis revealed that EnDeltaE47 preferentially forms heterodimers with myogenin. Interestingly, the chimeric repressor did not form DNA-binding heterodimers with MyoD in C2C12 myocytes. The failure to detect MyoD:EnDeltaE47 heterodimers in myoblasts was not due to protein conformational defects as both wild-type E47 and EnDeltaE47 readily formed DNA binding complexes with MyoD in vitro. These results indicate that E47 plays a crucial role in C2C12 myogenesis by serving as the preferred heterodimer partner of the myogenin protein. PMID- 11412047 TI - Stem cell factor-induced migration of mast cells requires p38 mitogen-activated protein kinase activity. AB - Stem cell factor (SCF) can be considered a cardinal cytokine in mast cell biology as it affects mast cell differentiation, survival, and migration. The objective of this study was to investigate the role of two mitogen-activated protein (MAP) kinases, extracellular signal-regulated kinase (ERK) and p38, in SCF-induced cell migration. This was examined in mouse mast cells by using PD 098059 and SB203580, which are specific inhibitors of mitogen-induced extracellular kinase (MEK) and p38 MAP kinase, respectively. SCF induced a rapid and transient activation of ERK and p38 in a dose-dependent manner. Inhibition of p38 activity by SB203580 was paralleled with a marked reduction of migration toward SCF, whereas the effect of the MEK inhibitor was less pronounced. This is the first report of a physiological function of SCF-dependent activation of p38. Whether p38-mediated mast cell migration is a possible target for suppression of mast cell hyperplasia remains to be determined. PMID- 11412048 TI - Incidence of malignant disease in biopsy-proven inflammatory myopathy. A population-based cohort study. AB - BACKGROUND: The validity and magnitude of an association between myositis and malignant disease continue to be debated. Such issues as the legitimacy of a myositis diagnosis and distinction among myositis subgroups in previous population-based studies remain unresolved. OBJECTIVE: To determine the risk for malignant disease in patients with biopsy-proven inflammatory myopathies. DESIGN: Population-based, retrospective cohort study. SETTING: Victoria, Australia. PATIENTS: 537 patients in whom a biopsy-positive idiopathic inflammatory myopathy was first diagnosed from 1981 through 1995. MEASUREMENTS: Standardized incidence ratios were calculated to compare the incidence of malignant disease in patients with inflammatory myopathy and the general population. RESULTS: A total of 116 cases of malignant disease were found in 104 patients. Seventy-four cases were identified concurrently with (within 7 days) or after diagnosis of myositis. The highest risk for malignant disease was associated with dermatomyositis (standardized incidence ratio, 6.2 [95% CI, 3.9 to 10.0]). The risk was also increased in polymyositis (standardized incidence ratio, 2.0 [CI, 1.4 to 2.7]), although the relative risk for malignant disease in dermatomyositis compared with polymyositis was 2.4 (CI, 1.3 to 4.2). An increased risk for malignant disease was also found in inclusion-body myositis (standardized incidence ratio, 2.4 [CI, 1.2 to 4.9]). The excess risk for malignant disease diminished with time (standardized incidence ratio, 4.4 [CI, 2.7 to 7.1] in the first year; 3.4 [CI, 2.3 to 5.1] between 1 and 3 years; 2.2 [CI, 1.3 to 3.9] between 3 and 5 years; and 1.6 [CI, 1.0 to 2.6] beyond 5 years [ P for trend, 0.002]). CONCLUSION: The risk for malignant disease is increased in biopsy-proven dermatomyositis and polymyositis and also appears to be increased in inclusion-body myositis. PMID- 11412049 TI - Prognostic disclosure to patients with cancer near the end of life. AB - BACKGROUND: Patients' understanding of their prognosis informs numerous medical and nonmedical decisions, but patients with cancer and their physicians often have disparate prognostic expectations. OBJECTIVE: To determine whether physician behavior might contribute to the disparity between patients' and physicians' prognostic expectations. DESIGN: Prospective cohort study. SETTING: Five hospices in Chicago, Illinois. PATIENTS: 326 patients with cancer. INTERVENTION: Physicians formulated survival estimates and also indicated the survival estimates that they would communicate to their patients if the patients insisted. MEASUREMENTS: Comparison of the formulated and communicated prognoses. RESULTS: For 300 of 311 evaluable patients (96.5%), physicians were able to formulate prognoses. Physicians reported that they would not communicate any survival estimate 22.7% (95% CI, 17.9% to 27.4%) of the time, would communicate the same survival estimate they formulated 37% (CI, 31.5% to 42.5%) of the time, and would communicate a survival estimate different from the one they formulated 40.3% (CI, 34.8% to 45.9%) of the time. Of the discrepant survival estimates, most (70.2%) were optimistically discrepant. Multivariate analysis revealed that older patients were more likely to receive frank survival estimates, that the most experienced physicians and the physicians who were least confident about their prognoses were more likely to favor no disclosure over frank disclosure, and that female physicians were less likely to favor frank disclosure over pessimistically discrepant disclosure. CONCLUSIONS: Physicians reported that even if patients with cancer requested survival estimates, they would provide a frank estimate only 37% of the time and would provide no estimate, a conscious overestimate, or a conscious underestimate most of the time (63%). This pattern may contribute to the observed disparities between physicians' and patients' estimates of survival. PMID- 11412050 TI - The effect of fruit and vegetable intake on risk for coronary heart disease. AB - BACKGROUND: Many constituents of fruits and vegetables may reduce the risk for coronary heart disease, but data on the relationship between fruit and vegetable consumption and risk for coronary heart disease are sparse. OBJECTIVE: To evaluate the association of fruit and vegetable consumption with risk for coronary heart disease. DESIGN: Prospective cohort study. SETTING: The Nurses' Health Study and the Health Professionals' Follow-Up Study. PARTICIPANTS: 84 251 women 34 to 59 years of age who were followed for 14 years and 42 148 men 40 to 75 years who were followed for 8 years. All were free of diagnosed cardiovascular disease, cancer, and diabetes at baseline. MEASUREMENTS: The main outcome measure was incidence of nonfatal myocardial infarction or fatal coronary heart disease (1127 cases in women and 1063 cases in men). Diet was assessed by using food frequency questionnaires. RESULTS: After adjustment for standard cardiovascular risk factors, persons in the highest quintile of fruit and vegetable intake had a relative risk for coronary heart disease of 0.80 (95% CI, 0.69 to 0.93) compared with those in the lowest quintile of intake. Each 1-serving/d increase in intake of fruits or vegetables was associated with a 4% lower risk for coronary heart disease (relative risk, 0.96 [CI, 0.94 to 0.99]; P = 0.01, test for trend). Green leafy vegetables (relative risk with 1-serving/d increase, 0.77 [CI, 0.64 to 0.93]), and vitamin C-rich fruits and vegetables (relative risk with 1-serving/d increase, 0.94 [CI, 0.88 to 0.99]) contributed most to the apparent protective effect of total fruit and vegetable intake. CONCLUSIONS: Consumption of fruits and vegetables, particularly green leafy vegetables and vitamin C-rich fruits and vegetables, appears to have a protective effect against coronary heart disease. PMID- 11412051 TI - Mediterranean and low-fat diets improve endothelial function in hypercholesterolemic men. AB - BACKGROUND: The regulatory function of the endothelium is altered in hypercholesterolemia, and the subsequent endothelial dysfunction plays a central role in the development of atherosclerosis. OBJECTIVE: To determine whether endothelial function in hypercholesterolemic patients is affected by replacing a saturated fat-enriched diet with a low-fat, low-saturated fat diet (the U.S. National Cholesterol Education Program stage 1 [NCEP-1] diet) or a diet rich in monounsaturated fat (such as that common in Mediterranean countries). DESIGN: Intervention dietary study with a baseline phase and two randomized crossover dietary periods. SETTING: Hospital Universitario Reina Sofia, Cordoba, Spain. PATIENTS: 22 hypercholesterolemic men. INTERVENTION: Patients followed a diet high in saturated fat, then were assigned in a crossover design to the NCEP-1 diet or a Mediterranean diet. Each dietary period lasted 28 days. MEASUREMENTS: Plasma P-selectin levels, lipid concentrations, and endothelial function. RESULTS: Compared with the saturated fat diet, flow-mediated dilatation increased during the Mediterranean diet but not during the NCEP-1 diet. In addition, levels of plasma cholesterol, low-density lipoprotein cholesterol, apolipoprotein B, and P-selectin decreased during the NCEP-1 and Mediterranean diets. CONCLUSION: In hypercholesterolemic men, diets low in fat (especially saturated fat) and diets rich in monounsaturated fats improve endothelial function. PMID- 11412052 TI - Preliminary evidence of impaired thinking in sick patients. AB - BACKGROUND: Earlier anecdotal observations suggested to us that certain aspects of judgment in sick adults approximate the thinking of children. OBJECTIVE: To describe changes in judgment associated with serious illness in otherwise competent adults. DESIGN: Cohort study. SETTING: Urban acute-care hospital and senior citizen center. PARTICIPANTS: Sicker (Karnofsky score 50; n = 39) hospitalized patients were compared with controls (n = 28). Normal performance on the Mini-Mental State Examination (score >/= 24) was required for study entrance. MEASUREMENTS: Seven Piagetian tasks of judgment designed to study childhood cognitive development. Degree of sickness was determined by using the Karnofsky scale of physical function. RESULTS: Patients with Karnofsky scores of 50 or less responded correctly to fewer Piagetian tasks than controls (mean [+/-SD], 1.8 +/- 2.6 vs. 5.9 +/- 1.6; P < 0.001). Furthermore, a smaller proportion of sicker patients responded correctly to each of the seven tasks. Patients with Karnofsky scores greater than 50 did not perform differently than controls. CONCLUSION: In sicker hospitalized patients, performance on seven Piagetian tasks of judgment was similar to that among children younger than 10 years of age. This evidence of cognitive impairment warrants further investigation. PMID- 11412053 TI - Methodologic pitfalls in the determination of genetic anticipation: the case of Crohn disease. AB - BACKGROUND: The term genetic anticipation is used when genetically transmitted disease manifests at increasingly younger ages with each succeeding generation: that is, if the offspring of patients develop the disease, they will tend to do so at an earlier age than their parents. In some monogenetic disorders, genetic anticipation has a biological basis in expanded genetic triplet repeats; however, some have claimed that it occurs in polygenic disorders, such as Crohn disease, in which its mechanism cannot be explained. OBJECTIVE: To show how apparent changes in age at diagnosis of Crohn disease between generations, which could suggest genetic anticipation, can be an artifact of inadequate analysis based on age at diagnosis in cohorts that have not been followed for a sufficiently long time. DESIGN: Comparison of ages at diagnosis of Crohn disease among different birth cohorts, before and after adjustment for observation time. SETTING: Meyerhoff Center for Inflammatory Bowel Disease, Johns Hopkins Hospital, Baltimore, Maryland. PATIENTS: 928 consecutive outpatients with Crohn disease. MEASUREMENTS: Trends in age at diagnosis of Crohn disease among birth cohorts were determined by calculating Pearson correlation coefficients and performing Kaplan-Meyer analysis before and after adjustment for observation time. Adjustment for observation time was performed by ensuring that the time during which all included patients were at risk for Crohn disease was equal and that all patients had developed disease by the end of the risk period. RESULTS: Mean age at diagnosis decreased by approximately 5 years with each subsequent 10-year birth cohort on the basis of crude cross-sectional data that could suggest genetic anticipation between generations. However, after adjustment for observation time, the age at diagnosis decreased minimally, if at all, with each successive generation. CONCLUSIONS: Apparent genetic anticipation can be explained by observational biases without invoking any additional genetic influences. Claims for genetic anticipation must be based on methods that properly account for the duration of observation in all persons being studied. PMID- 11412054 TI - Evolving indications for permanent pacemakers. AB - New indications for permanent cardiac pacing have been developed in recent years, with numerous studies demonstrating improved clinical outcomes in a variety of disorders. Because hypertrophic obstructive cardiomyopathy, dilated cardiomyopathy, heart failure, neurocardiogenic syncope, and atrial fibrillation are common conditions, every clinician should be aware of evolving alternative therapies for them. Observational studies in patients with refractory, symptomatic hypertrophic obstructive cardiomyopathy and significant left ventricular outflow gradient at rest suggest that cardiac pacing may result in symptomatic and hemodynamic improvement. Clinical trials have not shown conclusive evidence regarding the long-term benefit from pacing in these patients, and it is unclear whether pacing will be a preferred treatment option. Preliminary data suggest that pacing is a viable adjunctive therapeutic approach for improving symptoms in patients with dilated cardiomyopathy and heart failure. Mortality benefit has yet to be established, but it is to be hoped that ongoing randomized clinical trials will provide definitive information on that issue. Patients with refractory neurocardiogenic syncope or those who are intolerant of medical treatment may benefit from pacing therapies, especially those that use rate-drop sensor algorithms. Biatrial pacing has emerged as a technique that resynchronizes atrial electrical activity and has been shown to prevent atrial fibrillation. Multisite atrial pacing for the prevention of atrial fibrillation is considered investigational but seems promising. Newer indications for pacing are expected to result in improved clinical outcomes for hypertrophic obstructive cardiomyopathy, dilated cardiomyopathy and heart failure, neurocardiogenic syncope, and the prevention of atrial fibrillation. PMID- 11412056 TI - Editor's world. PMID- 11412055 TI - Truth in the most optimistic way. PMID- 11412057 TI - Blue light and milk. PMID- 11412058 TI - The efficacy of spiritual healing. PMID- 11412059 TI - Blinding in placebo-controlled trials. PMID- 11412060 TI - Blinding in placebo-controlled trials. PMID- 11412062 TI - Ethical principles for everyone. PMID- 11412063 TI - Ethical principles for everyone. PMID- 11412064 TI - Ethical principles for everyone. PMID- 11412065 TI - Ethical principles for everyone. PMID- 11412067 TI - Treatment of lipodystrophy with troglitazone. PMID- 11412069 TI - Tamoxifen-induced total alopecia. PMID- 11412070 TI - Immunologic reactions and hepatitis B vaccine. PMID- 11412071 TI - Long-lasting effectiveness of intravenous immunoglobulin in a patient with inclusion-body myositis. PMID- 11412072 TI - Rituximab for bone lymphoma. PMID- 11412075 TI - Medical marijuana: the continuing story. PMID- 11412077 TI - Molecular machines. PMID- 11412078 TI - The physics of molecular motors. AB - Molecular motors convert chemical energy into mechanical force and movement. Operating at energies just above those of the thermal bath, these motors experience large fluctuations, and their physical description must be necessarily stochastic. Here, motor operation is described as a biased diffusion on a potential energy surface defined by the interactions of the motor with its track and its fuel. These ideas are illustrated with a model of the rotary movement of the F(o) motor. PMID- 11412079 TI - Electronically transduced molecular mechanical and information functions on surfaces. AB - Supramolecular chemistry and nanotechnology, along with their use in the construction of functional assemblies and devices, have merged into a challenging field of study. The development of methodologies for the integration and interfacing of molecular building blocks with solid supports and electronic transducers is essential for this research. We address recent applications of molecular, macromolecular, and biomolecular substances in the organization of signal-activated, electronically transduced molecular architectures on electrode surfaces. Photonic, electronic, magnetic, and chemical stimuli are used to trigger the switchable functions of these systems, which demonstrate either mechanical (e.g., translocation) or computational (e.g., memory) functions and provide enlightening insight and directions for the future evolution of the field. PMID- 11412080 TI - Switching devices based on interlocked molecules. AB - An architectural rationale and an experimental program aimed at the development of molecular electronics switching devices for memory and computing applications are discussed. Two-terminal molecular switch tunnel junctions are identified as the critical device components of molecular electronics-based circuitry. They can be tiled in two dimensions and are tolerant of manufacturing defects. Singly and multiply configurable solid-state switching devices that are based upon electrochemically switchable molecular and supramolecular systems are discussed in terms of both the synthesis of the molecular components and the fabrication and performance of the devices. PMID- 11412081 TI - Artificial molecular-level machines: which energy to make them work? AB - The concept of machine can be extended to the molecular level by designing and synthesizing (supra)molecular species capable of performing mechanical movements. The energy needed to make a machine work can be supplied as chemical energy, electrical energy, or light. When a chemical "fuel" is used, waste products are formed, whereas this is not the case when suitable photochemical or electrochemical energy inputs are employed. A number of elementary functions performed by molecular-level machines are illustrated, and more complex ones are foreseen. PMID- 11412082 TI - Cyclodextrin-based molecular machines. AB - Cyclodextrins have been used as a cyclic component in the construction of supramolecular architectures. Recently they have been studied as a component in the construction of rotaxanes and catenanes. A cyclodextrin ring can translocate in some rotaxane and catenane structures. Therefore, much attention has been given to cyclodextrins as a component of molecular shuttles, motors, and machines. Attempts to design and synthesize molecular-level machines using cyclodextrins as a cyclic component are described. PMID- 11412083 TI - On the way to rotaxane-based molecular motors: studies in molecular mobility and topological chirality. AB - ATP synthase represents a machine at the molecular level which couples the rotation of an axle in a wheel with the endergonic production of ATP, the general source of chemical energy in the cell. The natural system prototypically bears all features of a macroscopic motor: a rotor within a stator held by a membrane and fueled by a difference in chemical potential in the form of a proton gradient combined with a machine for ATP production. The assembly of axle and wheel to a rotor device reminds one very much of a rotaxane. In this Account, we discuss some important features of motors and their (potential) realization in simpler artificial model systems, that is, the molecular mobility of mechanically bound molecules, the importance of chirality for unidirectional motion, the sources of energy for driving the rotation, and the potential of using membranes and surfaces for ordering a large number of devices to achieve macroscopic effects. PMID- 11412084 TI - Shuttles and muscles: linear molecular machines based on transition metals. AB - Transition-metal-containing rotaxanes can behave as linear motors at the molecular level. The molecules are set into motion either by an electrochemical reaction or using a chemical signal. In a first example, a simple rotaxane is described that consists of a ring threaded by a two-coordination-site axle. The ring contains a bidentate ligand, coordinated to a copper center. The axle incorporates both a bidentate and a terdentate ligand. By oxidizing or reducing the copper center to Cu(II) or Cu(I) respectively, the ring glides from a given position on the axle to another position and vice versa. By generalizing the concept to a rotaxane dimer, whose synthesis involves a quantitative double threading reaction triggered by copper(I) complexation, a molecular assembly reminiscent of a muscle is constructed. By exchanging the two metal centers of the complex (copper(I)/zinc(II)), a large-amplitude movement is generated, which corresponds to a contraction/stretching process. The copper(I)-containing rotaxane dimer is in a stretched situation (overall length approximately 8 nm), whereas the zinc(II) complexed compound is contracted (length approximately 6.5 nm). The stretching/contraction process is reversible and it is hoped that, in the future, other types of signals can be used (electrochemical or light pulse) to trigger the motion. PMID- 11412085 TI - Molecular machines based on metal ion translocation. AB - Transition metal ions can be moved reversibly between the two coordinatively unequivalent compartments A and B of a ditopic ligand, using as an input the variation of a bulk solution parameter, either pH or redox potential. In a redox driven translocation, the metal moves reversibly from A to B on cycling between two consecutive oxidation states (e.g., Cu(II)/Cu(I); Fe(III)/Fe(II)) by means of auxiliary oxidation and reduction reactions. In a pH-driven process, one compartment displays also acid-base properties (AH(n)() left arrow over right arrow A(n)(-) + nH(+)), and the M(n)(+) ion is translocated between B and A(n)(-) through consecutive addition of base and acid. PMID- 11412086 TI - Positive allosteric systems designed on dynamic supramolecular scaffolds: toward switching and amplification of guest affinity and selectivity. AB - Positive homotropic allosterism appears in important information transduction processes where chemical and physical signals are efficiently amplified. The phenomena are ubiquitous in nature, but the general methodology for the design of such allosteric systems is not yet established in an artificial system. This account reviews such artificial receptors that can bind guest ions and molecules in a positive allosteric manner and discusses what kinds of factors are indispensable as scaffolds in the design of this novel class of allosteric systems and what common factors are needed to realize the cooperativity. It has been shown that the scaffolds are mostly dynamic and are skillfully combined with the molecular recognition systems so that the subsequent guest binding can occur more favorably than the first guest binding. In addition, it has been suggested that positive homotropic allosterism can be utilized as a new strategy to attain high guest selectivity and guest affinity which cannot be attained by conventional 1:1-type guest binding. PMID- 11412087 TI - In control of motion: from molecular switches to molecular motors. AB - The design of molecular systems in which controlled linear and rotary motion can be achieved under the influence of an external signal is a major endeavor toward future nanoscale machinery. In this Account we describe the development of molecular switches and the discoveries that culminated in the first light-driven molecular motor. Various chiral optical molecular switches and their use as trigger elements to control organization and functions will be discussed. The construction of the first and second generation molecular motors is presented. PMID- 11412088 TI - Progress toward a rationally designed molecular motor. AB - A retrospective overview of the work in the author's laboratory leading to a prototype (43a) of a chemically powered molecular motor is provided. Beginning with a molecular brake (23) and then proceeding through a molecular "ratchet" (31), the development of a rationally designed molecular motor is described. The thermodynamic underpinnings of the concept are outlined, the synthetic routes to 23, 31, and 43a, are summarized, and the data documenting the function of 23, 31, and 43a are presented. PMID- 11412089 TI - Cancer isoform of a tumor-associated cell surface NADH oxidase (tNOX) has properties of a prion. AB - We have described a drug-responsive form of a cell surface NADH oxidase (hydroquinone oxidase) of cancer cells (tNOX) that exhibits unusual characteristics including resistance to proteases, resistance to cyanogen bromide digestion, and an ability to form amyloid filaments closely resembling those of spongiform encephalopathies and all of which are characteristics of PrP(sc) (PrP(res)), the presumed infective and proteinase K resistant particle of the scrapie prion. The tNOX protein from the HeLa cell surface copurified with authentic glyceraldehyde-3-phosphate dehydrogenase (muscle form) (GAPDH). Surprisingly, the tNOX-associated muscle GAPDH also was proteinase K resistant. In this paper, we show that combination of authentic rabbit muscle GAPDH with tNOX renders the GAPDH resistant to proteinase K digestion. This property, that of converting the normal form of a protein into a likeness of itself, is one of the defining characteristics of the group of proteins designated as prions. PMID- 11412090 TI - Catalytic acid-base groups in yeast pyruvate decarboxylase. 1. Site-directed mutagenesis and steady-state kinetic studies on the enzyme with the D28A, H114F, H115F, and E477Q substitutions. AB - The roles of four of the active center groups with potential acid-base properties in the region of pH optimum of pyruvate decarboxylase from Saccharomyces cerevisiae have been studied with the substitutions Asp28Ala, His114Phe, His115Phe, and Glu477Gln, introduced by site-directed mutagenesis methods. The steady-state kinetic constants were determined in the pH range of activity for the enzyme. The substitutions result in large changes in k(cat) and k(cat)/S(0.5) (and related terms), indicating that all four groups have a role in transition state stabilization. Furthermore, these results also imply that all four are involved in some manner in stabilizing the rate-limiting transition state(s) both at low substrate (steps starting with substrate binding and culminating in decarboxylation) and at high substrate concentration (steps beginning with decarboxylation and culminating in product release). With the exception of some modest effects, the shapes of neither the bell-shaped k(cat)/S(0.5)-pH (and related functions) plots nor the k(cat)-pH plots are changed by the substitutions. Yet, the fractional activity still remaining after substitutions virtually rules out any of the four residues as being directly responsible for initiating the catalytic process by ionizing the C2H. There is no effect on the C2 H/D exchange rate exhibited by the D28A and E477Q substitutions. These results strongly imply that the base-induced deprotonation at C2 is carried out by the only remaining base, the iminopyrimidine tautomer of the coenzyme, via intramolecular proton abstraction. The first product is released as CO(2) rather than HCO(3)(-) by both wild-type and E477Q and D28A variants, ruling out several mechanistic alternatives. PMID- 11412091 TI - Catalytic acid-base groups in yeast pyruvate decarboxylase. 2. Insights into the specific roles of D28 and E477 from the rates and stereospecificity of formation of carboligase side products. AB - Yeast pyruvate decarboxylase (YPDC), in addition to forming its metabolic product acetaldehyde, can also carry out carboligase reactions in which the central enamine intermediate reacts with acetaldehyde or pyruvate (instead of the usual proton electrophile), resulting in the formation of acetoin and acetolactate, respectively (typically, 1% of the total reaction). Due to the common mechanism shared by the acetaldehyde-forming and carboligase reactions through decarboxylation, a detailed analysis of the rates and stereochemistry of the carboligase products formed by the E477Q, D28A, and D28N active center YPDC variants was undertaken. While substitution at either position led to an approximately 2-3 orders of magnitude lower catalytic efficiency in acetaldehyde formation, the rate of acetoin formation by the E477Q and D28N variants was higher than that by wild-type enzyme. Comparison of the steady-state data for acetaldehyde and acetoin formation revealed that the rate-limiting step for acetaldehyde formation by the D28A, H114F, H115F, and E477Q variants is a step post-decarboxylation. In contrast to the wild-type YPDC and the E477Q variant, the D28A and D28N variants could synthesize acetolactate as a major product. The lower overall rate of side-product formation by the D28A variant than wild-type enzyme attests to participation of D28 in steps leading up to and including decarboxylation. The results also provide insight into the state of ionization of the side chains examined. (R)-Acetoin is produced by the variants with greater enantiomeric excess than by wild-type YPDC. (S)-Acetolactate is the predominant enantiomer produced by the D28-substituted variants, the same configuration as produced by the related plant acetolactate synthase. PMID- 11412092 TI - Catalytic acid-base groups in yeast pyruvate decarboxylase. 3. A steady-state kinetic model consistent with the behavior of both wild-type and variant enzymes at all relevant pH values. AB - The widely quoted kinetic model for the mechanism of yeast pyruvate decarboxylase (YPDC, EC 4.1.1.1), an enzyme subject to substrate activation, is based on data for the wild-type enzyme under optimal experimental conditions. The major feature of the model is the obligatory binding of substrate in the regulatory site prior to substrate binding at the catalytic site. The activated monomer would complete the cycle by irreversible decarboxylation of the substrate and product (acetaldehyde) release. Our recent kinetic studies of YPDC variants substituted at positions D28 and E477 at the active center necessitate some modification of the mechanism. It was found that enzyme without substrate activation apparently is still catalytically competent. Further, substrate-dependent inhibition of D28 substituted variants leads to an enzyme form with nonzero activity at full saturation, requiring a second major branch point in the mechanism. Kinetic data for the E477Q variant suggest that three consecutive substrate binding steps may be needed to release product acetaldehyde, unlikely if YPDC monomer is the minimal catalytic unit with only two binding sites for substrate. A model to account for all kinetic observations involves a functional dimer operating through alternation of active sites. In the context of this mechanism, roles are suggested for the active center acid-base groups D28, E477, H114, and H115. The results underline once more the enormous importance that both aromatic rings of the thiamin diphosphate, rather than only the thiazolium ring, have in catalysis, a fact little appreciated prior to the availability of the 3-dimensional structure of these enzymes. PMID- 11412093 TI - Influence of the aglycone region of the substrate binding cleft of Pseudomonas xylanase 10A on catalysis. AB - Pseudomonas cellulosa xylanase 10A (Pc Xyn10A) contains an extended substrate binding cleft comprising three glycone (-1 to -3) and four aglycone (+1 to +4) subsites and, typical of retaining glycoside hydrolases, exhibits transglycosylation activity at elevated substrate concentrations. In a previous study [Charnock, S. J., et al. (1997) J. Biol. Chem. 272, 2942-2951], it was demonstrated that the -2 subsite mutations E43A and N44A caused a 100-fold reduction in activity against xylooligosaccharides, but did not influence xylanase activity. This led to the proposal that the low activity of these mutants against xylooligosaccharides was due to nonproductive complex formation between these small substrates and the extended aglycone region of the active site. To test this hypothesis, key residues at the +2 (Asn182), +3 (Tyr255), and +4 (Tyr220) subsites were substituted for alanine, and the activity of the mutants against polysaccharides and oligosaccharides was evaluated. All the aglycone mutants exhibited greatly reduced or no transglycosylating activity, and the triple mutants, E43A/Y220A/Y255A and E43A/N182A/Y255A, had activity against xylotriose similar to that of E43A. The aglycone mutations caused an increase in both k(cat) and K(m) against xylan, with N182A/Y220A/Y255A and N182A/Y255A exhibiting 25- and 15-fold higher k(cat) values, respectively, than wild-type Pc Xyn10A. These data indicate that Glu43 plays a role in binding xylooligosaccharides, but not xylan, suggesting that the mechanisms by which Pc Xyn10A binds polysaccharides and oligosaccharides are distinct. The increased k(cat) of the mutants against xylan indicates that the aglycone region of wild type Pc Xyn10A restricts the rate of catalysis by limiting diffusion of the cleaved substrate, generated at the completion of the k(2) step, out of the active site. PMID- 11412094 TI - Mutations in yeast protein phosphatase type 1 that affect targeting subunit binding. AB - Protein phosphatase type 1 (PP1) is a major Ser/Thr protein phosphatase that is involved in many cellular processes. The activity of PP1 is controlled by regulatory subunits, many of which are thought to bind to a hydrophobic groove in PP1 via a short consensus sequence termed the V/IXF motif. To test this hypothesis, 11 variants of yeast PP1 (Glc7) were constructed in which one or more of the residues comprising the groove were changed to alanine. These variants were tested for their biological activity in vivo, for their biochemical activity in vitro, and for their ability to associate with three PP1 binding proteins. Five variants are unable to complement the essential function of PP1 in vivo although they are catalytically active in vitro. Many of the mutants are deficient in binding two V/IXF-containing subunits, Gac1 and Reg1, which regulate glycogen accumulation and glucose repression, respectively, but all retain the ability to associate with Sds22, a regulatory subunit that lacks this motif. The subcellular locations at which PP1 normally accumulates (bud neck, nucleolus, spindle pole body) were not occupied by one PP1 variant. Additionally, we provide evidence that mutations in the hydrophobic groove of PP1 affect substrate specificity. Together, these results demonstrate the importance of the hydrophobic groove for the interaction with regulatory subunits, for the proper subcellular localization of PP1 and for the substrate specificity of PP1. PMID- 11412095 TI - Beta D305A mutant of tryptophan synthase shows strongly perturbed allosteric regulation and substrate specificity. AB - Substrate channeling in the tryptophan synthase bienzyme is regulated by allosteric interactions. Allosteric signals are transmitted via a scaffolding of structural elements that includes a monovalent cation-binding site and salt bridging interactions between the side chains of betaAsp 305, betaArg 141, betaLys 167, and alphaAsp 56 that appear to modulate the interconversion between open and closed conformations. betaAsp 305 also interacts with the hydroxyl group of the substrate L-Ser in some structures. One possible functional role for betaAsp 305 is to ensure the allosteric transmission that triggers the switching of alphabeta-dimeric units between open and closed conformations of low and high activity. This work shows that substitution of betaAsp 305 with Ala (betaD305A) decreases the affinity of the beta-site for the substrate L-Ser, destabilizes the enzyme-bound alpha-aminoacrylate, E(A-A), and quinonoid species, E(Q), and changes the nucleophile specificity of the beta-reaction. The altered specificity provides a biosynthetic route for new L-amino acids derived from substrate analogues. betaD305A also shows an increased rate of formation of pyruvate upon reaction with L-Ser relative to the wild-type enzyme. The formation of pyruvate is strongly inhibited by the binding of benzimidazole to E(A-A). Upon reaction with L-Ser and in the presence of the alpha-site substrate analogue, alpha glycerol phosphate, the Na(+) form of betaD305A undergoes inactivation via reaction of nascent alpha-aminoacrylate with bound PLP. This work establishes important roles for betaAsp 305 both in the conformational change between open and closed states that takes place at the beta-site during the formation of the E(A-A) and in substrate binding and recognition. PMID- 11412096 TI - Does "butyrylization" of acetylcholinesterase through substitution of the six divergent aromatic amino acids in the active center gorge generate an enzyme mimic of butyrylcholinesterase? AB - The active center gorge of human acetylcholinesterase (HuAChE) is lined by 14 aromatic residues, whereas in the closely related human butyrylcholinesterase (HuBChE) 3 of the aromatic active center residues (Phe295, Phe297, Tyr337) as well as 3 of the residues at the gorge entrance (Tyr72, Tyr124, Trp286) are replaced by aliphatic amino acids. To investigate whether this structural variability can account for the reactivity differences between the two enzymes, gradual replacement of up to all of the 6 aromatic residues in HuAChE by the corresponding residues in HuBChE was carried out. The affinities of the hexamutant (Y72N/Y124Q/W286A/F295L/F297V/Y337A) toward tacrine, decamethonium, edrophonium, huperzine A, or BW284C51 differed by about 5-, 80-, 170-, 25000-, and 17000-fold, respectively, from those of the wild-type HuAChE. For most of these prototypical noncovalent active center and peripheral site ligands, the hexamutant HuAChE displayed a reactivity phenotype closely resembling that of HuBChE. These results support the accepted view that the active center architectures of AChE and BChE differ mainly by the presence of a larger void space in BChE. Nevertheless, reactivity of the hexamutant HuAChE toward the substrates acetylthiocholine and butyrylthiocholine, or covalent ligands such as phosphonates and the transition state analogue m-(N,N,N trimethylammonio)trifluoroacetophenone (TMTFA), is about 45-170-fold lower than that of HuBChE. Most of this reduction in reactivity can be related to the combined replacements of the three aromatic residues at the active center, Phe295, Phe297, and Tyr337. We propose that the hexamutant HuAChE, unlike BChE, is impaired in its capacity to accommodate certain tetrahedral species in the active center. This impairment may be related to the enhanced mobility of the catalytic histidine His447, which is observed in molecular dynamics simulations of the hexamutant and the F295L/F297V/Y337A HuAChE enzymes but not in the wild type HuAChE. PMID- 11412097 TI - Characterization of the allosteric anion-binding site of O-acetylserine sulfhydrylase. AB - A new crystal structure of the A-isozyme of O-acetylserine sulfhydrylase-A (OASS) with chloride bound to an allosteric site located at the dimer interface has recently been determined [Burkhard, P., Tai, C.-H., Jansonius, J. N., and Cook, P. F. (2000) J. Mol. Biol. 303, 279-286]. Data have been obtained from steady state and presteady-state kinetic studies and from UV-visible spectral studies to characterize the allosteric anion-binding site. Data obtained with chloride and sulfate as inhibitors indicate the following: (i) chloride and sulfate prevent the formation of the external aldimines with L-cysteine or L-serine; (ii) chloride and sulfate increase the external aldimine dissociation constants for O acetyl-L-serine, L-methionine, and 5-oxo-L-norleucine; (iii) chloride and sulfate bind to the allosteric site in the internal aldimine and alpha-aminoacrylate external aldimine forms of OASS; (iv) sulfate also binds to the active site. Sulfide behaves in a manner identical to chloride and sulfate in preventing the formation of the L-serine external aldimine. The binding of chloride to the allosteric site is pH independent over the pH range 7-9, suggesting no ionizable enzyme side chains ionize over this pH range. Inhibition by sulfide is potent (K(d) is 25 microM at pH 8) suggesting that SH(-) is the physiologic inhibitory species. PMID- 11412098 TI - Structures of Escherichia coli branched-chain amino acid aminotransferase and its complexes with 4-methylvalerate and 2-methylleucine: induced fit and substrate recognition of the enzyme. AB - The following three-dimensional structures of three forms of Escherichia coli branched-chain amino acid aminotransferase (eBCAT) have been determined by the X ray diffraction method: the unliganded pyridoxal 5'-phosphate (PLP) form at a 2.1 A resolution, and the two complexes with the substrate analogues, 4 methylvalerate (4-MeVA) as the Michaelis complex model and 2-methylleucine (2 MeLeu) as the external aldimine model at 2.4 A resolution. The enzyme is a trimer of dimers, and each subunit consists of small and large domains, and the interdomain loop. The active site is formed by the residues at the domain interface and those from two loops of the other subunit of the dimer unit, and binds one PLP with its re-face directed toward the protein side. Upon binding of a substrate, Arg40 changes its side-chain direction to interact with the interdomain loop, and the loop, which is disordered in the unliganded form, shows its ordered structure on the active-site cavity, interacts with the hydrophobic side chain of the substrate, and shields it from the solvent region. The substrate binds to the active-site pocket with its alpha-hydrogen toward the protein side, its side-chain on the side of O3 of PLP, and its alpha-carboxylate on the side of the phosphate group of PLP. The hydrophobic side-chain of the substrate is recognized by Phe36, Trp126, Tyr129, Tyr164, Tyr31*, and Val109*. The alpha-carboxylate of the substrate binds to the unique site constructed by three polar groups (two main-chain NH groups of the beta-turn at Thr257 and Ala258 and the hydroxy group of Tyr95) which are activated by the access of Arg40 to the main-chain C=O group of the beta-turn and the coordination of Arg97 to the hydroxy group. Since Arg40 is the only residue that significantly changes its side-chain conformation and directly interacts with the interdomain loop and the beta-turn, the residue plays important roles in the induced fit of the interdomain loop and the alpha-carboxylate recognition of the substrate. PMID- 11412099 TI - A functioning chimera of the cyclic nucleotide-binding domain from the bovine retinal rod ion channel and the DNA-binding domain from catabolite gene activating protein. AB - The eukaryotic cyclic nucleotide-gated (CNG) ion channels are a family of large membrane proteins activated by cytoplasmic cGMP or cAMP. Their cyclic nucleotide binding domain is structurally homologous with that of the catabolite gene activator protein (CAP), a soluble Escherichia coli transcription factor. Differences in ligand activation among sensory channels suggest differences in the underlying molecular mechanisms of signal readout. To study the structural, functional, and conformational consequences of nucleotide binding, we fused the cyclic nucleotide-binding domain from the bovine retinal rod CNG channel alpha subunit (Bralpha) to the DNA-binding domain from CAP. The chimera forms a soluble dimer that binds both cGMP and cAMP with association constants of 3.7 x 10(4) M( 1) for [(3)H]cGMP and 3.1 x 10(4) M(-1) for [(3)H]cAMP. The binding of cAMP, but not cGMP, exposes a chymotrypsin cleavage site in the chimera at a position similar to the site in the CAP exposed by cAMP binding. At high cAMP concentrations, a biphasic pattern of cleavage is seen, suggesting that the low affinity cAMP binding sites are also occupied. Cyclic AMP promotes specific binding to a DNA fragment encoding the lac operator region; the K(d) for the protein-DNA binding is approximately 200 nM, which is 2-fold higher than the K(d) for CAP under identical conditions. A 7 A crystal structure shows that the overall secondary and tertiary structure of Bralpha/CAP is the same as that of CAP with two cAMP molecules bound per dimer. The biochemical characterization of the chimera suggests it will be a useful system for testing hypotheses about channel activation, providing further insight into channel function. PMID- 11412100 TI - The arginine finger loop of yeast and human GAP is a determinant for the specificity toward Ras GTPase. AB - In this work, we have studied the role of the arginine finger region in determining the specificity of the GTPase activating proteins (GAPs) Saccharomyces cerevisiae Ira2p and human p120-GAP toward yeast Ras2p and human Ha Ras p21. It is known that p120-GAP can enhance both Ras2p and Ha-Ras GTPase activities, whereas Ira2p is strictly specific for Ras2p and fails to activate Ha Ras GTPase. Substitution in Ira2p of the arginine following the arginine finger with alanine, the residue found in the corresponding position of p120-GAP, or by glycine as found in neurofibromin, evokes a low but significant stimulation of Ha Ras GTPase. The stimulatory activity of Ira2p on Ha-Ras increased by substituting segments of the finger loop region with p120-GAP residues, especially with the six residues forming the tip of the arginine loop. In p120-GAP, substitution of the entire finger loop with the corresponding region of Ira2p led to a construct completely inactive on Ha-Ras GTPase but active on yeast Ras2p GTPase. Analysis of these results and modeling of Ira2p.Ras complexes emphasize the importance of the finger loop region not only for the catalytic activity but also as a structural determinant involved in the specificity of GAPs toward Ras proteins from different organisms. PMID- 11412101 TI - Mutational analysis of the K+-competitive inhibitor site of gastric H,K-ATPase. AB - The gastric H,K-ATPase is inhibited selectively and K(+)-competitively from its luminal surface by protonated imidazo[1,2alpha]pyridines (e.g., SCH28080). Identification of the amino acids in the membrane domain that affect SCH28080 inhibition should provide a template for modeling a luminally directed vestibule in this enzyme, based on the crystal structure of the sr Ca-ATPase. Five conserved carboxylic residues, Glu343, Glu795, Glu820, Asp824, Glu936, and unique Lys791 in the H,K-ATPase were mutated, and the effects of mutations on the K(i) for SCH28080, V(max), and K(m,app)[NH(4)(+)] were measured. A kinetic analysis of the ATP hydrolysis data indicated that all of these residues significantly affect the interaction of NH(4)(+) ions with the protein but only three of them, Glu795, Glu936, and Lys791, greatly affected SCH28080 inhibition. A Glu795Asp mutation increased the K(i) from 64 +/- 11 to 700 +/- 110 nM. Since, however, the mutation Glu795Gln did not change the K(i) (86 +/- 31 nM), this site has a significant spatial effect on inhibitor kinetics. A Glu936Asp mutation resulted in noncompetitive kinetics while Gln substitution had no effect either on inhibitor affinity or on the nature of the kinetics, suggesting that the length of the Glu936 side chain is critical for the exclusive binding of the ion and SCH28080. Mutation of Lys791 to Ser, the residue present in the SCH28080-insensitive Na,K ATPase, resulted in a 20-fold decrease in SCH28080 affinity, suggesting an important role of this residue in SCH28080 selectivity of the H,K-ATPase versus Na,K-ATPase. Mutations of Asp824, Glu343, and Glu820 increased the K(i) 2-3-fold, implying a relatively minor role for these residues in SCH28080 inhibition. It appears that the imidazopyridine moiety of SCH28080 in the protonated state interacts with residues near the negatively charged residues of the empty ion site from the luminal side (TM4, -5, -6, and -8) while the hydrophobic phenyl ring interacts with TM1 or TM2 (the latter conclusion based on previous data from photoaffinity labeling). The integrity of the SCH28080 binding site depends on the presence of Lys791, Glu936, and Glu795 in H,K-ATPase. A computer-generated model of this region illustrates the possible involvement of the residues previously shown to affect SCH28080 inhibition (Cys813, Ile816, Thr823, Met334, Val337) and may predict other residues that line the SCH28080 binding vestibule in the E(2) conformation of the pump. PMID- 11412102 TI - Fast deuterium access to the buried magnesium/manganese site in cytochrome c oxidase. AB - The recently determined crystal structures of bacterial and bovine cytochrome c oxidases show an area of organized water within the protein immediately above the active site where oxygen chemistry occurs. A pathway for exit of protons or water produced during turnover is suggested by possible connections of this aqueous region to the exterior surface. A non-redox-active Mg(2+) site is located in the interior of this region, and our previous studies [Florens, L., Hoganson, C., McCracken, J., Fetter, J., Mills, D., Babcock, G. T., and Ferguson-Miller, S. (1998) in Phototropic Prokaryotes (Peschek, G. A., Loeffelhard, W., and Schmetterer, G., Eds.) Kluwer Academic/Plenum, New York] have shown that the protons of water molecules that coordinate the metal can be exchanged within minutes of mixing with (2)H(2)O. Here we examine the extent and rate of deuterium exchange, using a combination of rapid freeze-quench and electron spin echo envelope modulation (ESEEM) analysis of Mn(2+)-substituted cytochrome c oxidase, which retains full activity. In the oxidized enzyme at room temperature, deuterium exchange at the Mn(2+) site occurs in less than 11 ms, which corresponds to an apparent rate constant higher than 3000 s(-1). The extent of deuterium substitution is dependent on the concentration of (2)H(2)O in the sample, indicative of rapid equilibrium, with three inner sphere (2)H(2)O exchanged per Mn(2+). This indicates that the water ligands of the Mn(2+)/Mg(2+) site, or the protons of these waters, can exchange with bulk solvent at a rate consistent with a role for this region in product release during turnover. PMID- 11412103 TI - NMR analysis of structure and dynamics of the cytosolic tails of integrin alpha IIb beta 3 in aqueous solution. AB - The structural and dynamic properties of the cytosolic tails of the adhesion receptor integrin alphaIIbbeta3, fused to a coiled-coil construct via (Gly)(3) linkers, were studied in aqueous solution by nuclear magnetic resonance (NMR) spectroscopy. Both tails were largely flexible and unstructured, although, in the beta3 tail, residues Arg(724)-Ala(735) have a propensity to form a helical structure and residues Asn(744)-Tyr(747) (NPLY) have a propensity to adopt reverse-turn conformations. The mutation beta3(Y747A) disrupted this reverse-turn tendency and markedly reduced the affinity of the head domain of the cytoskeletal protein, talin for the beta3 tail. Omission of the (Gly)(3) linker connecting the coiled-coiled helices and the integrin tails lead to helix propagation into the beta3 tail extending up to eight residues. A variety of different tail constructs were made and studied to reveal tail-tail interactions, but surprisingly no significant interactions between both tails could be detected within the context of our constructs. These results provide structural insight into a highly conserved beta tail motif (NPXY/F) required for integrin signaling and highlight a second transiently structured region (residues Arg(724)-Ala(735)), which might also be of functional significance. PMID- 11412104 TI - Structural and functional characterization of second-coordination sphere mutants of soybean lipoxygenase-1. AB - Lipoxygenases are an important class of non-heme iron enzymes that catalyze the hydroperoxidation of unsaturated fatty acids. The details of the enzymatic mechanism of lipoxygenases are still not well understood. This study utilizes a combination of kinetic and structural probes to relate the lipoxygenase mechanism of action with structural modifications of the iron's second coordination sphere. The second coordination sphere consists of Gln(495) and Gln(697), which form a hydrogen bond network between the substrate cavity and the first coordination sphere (Asn(694)). In this investigation, we compared the kinetic and structural properties of four mutants (Q495E, Q495A, Q697N, and Q697E) with those of wild type soybean lipoxygenase-1 and determined that changes in the second coordination sphere affected the enzymatic activity by hydrogen bond rearrangement and substrate positioning through interaction with Gln(495). The nature of the C-H bond cleavage event remained unchanged, which demonstrates that the mutations have not affected the mechanism of hydrogen atom tunneling. The unusual and dramatic inverse solvent isotope effect (SIE) observed for the Q697E mutant indicated that an Fe(III)-OH(-) is the active site base. A new transition state model for hydrogen atom abstraction is proposed. PMID- 11412105 TI - Preparation of potent cytotoxic ribonucleases by cationization: enhanced cellular uptake and decreased interaction with ribonuclease inhibitor by chemical modification of carboxyl groups. AB - Carboxyl groups of bovine RNase A were amidated with ethylenediamine (to convert negative charges of carboxylate anions to positive ones), 2-aminoethanol (to eliminate negative charges), and taurine (to keep negative charges), respectively, by a carbodiimide reaction. Human RNase 1 was also modified with ethylenediamine. Surprisingly, the modified RNases were all cytotoxic toward 3T3 SV-40 cells despite their decreased ribonucleolytic activity. However, their enzymatic activity was not completely eliminated by the presence of excess cytosolic RNase inhibitor (RI). As for native RNase A and RNase 1 which were not cytotoxic, they were completely inactivated by RI. More interestingly, within the cytotoxic RNase derivatives, cytotoxicity correlated well with the net positive charge. RNase 1 and RNase A modified with ethylenediamine were more cytotoxic than naturally occurring cytotoxic bovine seminal RNase. An experiment using the fluorescence-labeled RNase derivatives indicated that the more cationic RNases were more efficiently adsorbed to the cells. Thus, it is suggested that the modification of carboxyl groups could change complementarity of RNase to RI and as a result endow RNase cytotoxicity and that cationization enhances the efficiency of cellular uptake of RNase so as to strengthen its cytotoxicity. The finding that an extracellular human enzyme such as RNase 1 could be effectively internalized into the cell by cationization suggests that cationization is a simple strategy for efficient delivery of a protein into cells and may open the way of the development of new therapeutics. PMID- 11412106 TI - "Opening" the ferritin pore for iron release by mutation of conserved amino acids at interhelix and loop sites. AB - Ferritin concentrates, stores, and detoxifies iron in most organisms. The iron is a solid, ferric oxide mineral (< or =4500 Fe) inside the protein shell. Eight pores are formed by subunit trimers of the 24 subunit protein. A role for the protein in controlling reduction and dissolution of the iron mineral was suggested in preliminary experiments [Takagi et al. (1998) J. Biol. Chem. 273, 18685-18688] with a proline/leucine substitution near the pore. Localized pore disorder in frog L134P crystals coincided with enhanced iron exit, triggered by reduction. In this report, nine additional substitutions of conserved amino acids near L134 were studied for effects on iron release. Alterations of a conserved hydrophobic pair, a conserved ion pair, and a loop at the ferritin pores all increased iron exit (3-30-fold). Protein assembly was unchanged, except for a slight decrease in volume (measured by gel filtration); ferroxidase activity was still in the millisecond range, but a small decrease indicates slight alteration of the channel from the pore to the oxidation site. The sensitivity of reductive iron exit rates to changes in conserved residues near the ferritin pores, associated with localized unfolding, suggests that the structure around the ferritin pores is a target for regulated protein unfolding and iron release. PMID- 11412107 TI - Laser-induced photo-cross-linking of cisplatin-modified DNA to HMG-domain proteins. AB - Laser-induced photo-cross-linking was investigated for DNA, modified with cisplatin at specific sites, bound to structure-specific recognition domains of proteins in the high-mobility group (HMG) class. The efficiency of photo-cross linking depends on the wavelength and power of the laser, the nature of the protein domain, and the oligodeoxyribonucleotide sequences flanking the platinated site. Introduction of 5-iodouridine at thymine sites of the oligodeoxyribonucleotide as an additional photoreactive group did not increase the photo-cross-linking yield. Formation of platinum-mediated DNA-DNA interstrand cross-linking observed previously upon irradiation with 302 nm light [Kane, S. A., and Lippard, S. J. (1996) Biochemistry 35, 2180-2188] was significantly reduced with laser irradiation. HMG1 domain B is superior to domain A for platinum-mediated photo-cross-linking, a result attributed to the different positioning of the proteins with respect to the platinum adduct and the greater ability of domain B to access photolabilized platinum in the major groove. Studies with proteins containing specifically mutated amino acids, and with DNA probes in which the sequences flanking the platinum cross-link site were varied, suggest that the most effective photo-cross-linking occurs for protein domains bound symmetrically and flexibly to cisplatin-modified DNA. The thermodynamic equilibrium between the protein-platinated DNA complex and its components, revealed in gel electrophoretic mobility shift assays (EMSAs), is significantly shifted to the right upon irreversible photo-cross-linking. Thus, only upon photo cross-linking can the interaction of cisplatin-DNA 1,3-intrastrand d(GpTpG) or interstrand cross-links with HMG1 domain B protein be detected. Photo-cross linking is thus an effective tool for investigating the interaction of cisplatin modified DNA with damage-recognition proteins under heterogeneous conditions such those in cell extracts or living cells. PMID- 11412108 TI - Role of the ATP synthase alpha-subunit in conferring sensitivity to tentoxin. AB - Tentoxin, produced by phytopathogenic fungi, selectively affects the function of the ATP synthase enzymes of certain sensitive plant species. Binding of tentoxin to a high affinity (K(i) approximately 10 nM) site on the chloroplast F(1) (CF(1)) strongly inhibits catalytic function, whereas binding to a second, lower affinity site (K(d) > 10 microM) leads to restoration and even stimulation of catalytic activity. Sensitivity to tentoxin has been shown to be due, in part, to the nature of the amino acid residue at position 83 on the catalytic beta subunit of CF(1). An aspartate in this position is required, but is not sufficient, for tentoxin inhibition. By comparison with the solved structure of mitochondrial F(1) [Abrahams, J. P., Leslie, A. G. W., Lutter, R., and Walker, J. E. (1994) Nature 370, 621-628], Asp83 is probably located at an interface between alpha and beta subunits on CF(1) where residues on the alpha subunit could also participate in tentoxin binding. A hybrid core F(1) enzyme assembled with beta and gamma subunits of the tentoxin-sensitive spinach CF(1), and an alpha subunit of the tentoxin-insensitive photosynthetic bacterium Rhodospirillum rubrum F(1) (RrF(1)), was stimulated but not inhibited by tentoxin [Tucker, W. C., Du, Z., Gromet-Elhanan, Z. and Richter, M. L. (2001) Eur. J. Biochem. 268, 2179-2186]. In this study, chimeric alpha subunits were prepared by introducing short segments of the spinach CF(1) alpha subunit from a poorly conserved region which is immediately adjacent to beta-Asp83 in the crystal structure, into equivalent positions in the RrF(1) alpha subunit using oligonucleotide-directed mutagenesis. Hybrid enzymes containing these chimeric alpha subunits had both the high affinity inhibitory tentoxin binding site and the lower affinity stimulatory site. Changing beta-Asp83 to leucine resulted in loss of both inhibition and stimulation by tentoxin in the chimeras. The results indicate that tentoxin inhibition requires additional alpha residues that are not present on the RrF(1) alpha subunit. A structural model of a putative inhibitory tentoxin binding pocket is presented. PMID- 11412109 TI - 3-Methyleneoxindole: an affinity label of glutathione S-transferase pi which targets tryptophan 38. AB - The compound 3-methyleneoxindole (MOI), a photooxidation product of the plant auxin indole-3-acetic acid, functions as an affinity label of the dimeric pi class glutathione S-transferase (GST) isolated from pig lung. MOI inactivates the enzyme to a limit of 14% activity. The k for inactivation by MOI is decreased 20 fold by S-hexylglutathione but only 2-fold by S-methylglutathione, suggesting that MOI does not react entirely within the glutathione site. The striking protection against inactivation provided by S-(hydroxyethyl)ethacrynic acid indicates that MOI reacts in the active site region involving both the glutathione and the xenobiotic substrate sites. Incorporation of [(3)H]MOI up to approximately 1 mol/mol of enzyme dimer concomitant with maximum inactivation suggests that there are interactions between subunits. Fractionation of the proteolytic digest of [(3)H]MOI-modified GST pi yielded Trp38 as the only labeled amino acid. The crystal structure of the human GST pi-ethacrynic acid complex (2GSS) shows that the indole of Trp38 is less than 4 A from ethacrynic acid. Similarly, MOI may bind in this substrate site. In contrast to its effect on the pi class GST, MOI inactivates much less rapidly and extensively alpha and mu class GSTs isolated from the rat. These results show that MOI reacts preferentially with GST pi. Such a compound may be useful in novel combination chemotherapy to enhance the efficacy of alkylating cancer drugs while minimizing toxic side effects. PMID- 11412110 TI - Comparative binding properties of metallobleomycins with DNA 10-mers. AB - Properties of the interaction of bleomycin (Blm) and metallobleomycins [M = Zn, Cu(II), Fe(III), and HO(2)-Co(III)] with site-specific and nonspecific DNA oligomers, d(GGAAGCTTCC)(2) (I) and d(GGAAATTTCC)(2) (II), respectively, were investigated. With both 10-mers association constants increased in the series Blm A(2), ZnBlm A(2), Cu(II)Blm A(2), Fe(III)Blm A(2), and HO(2)-Co(III)Blm A(2). Generally, the metallobleomycins were bound with a modestly higher affinity to I. One-dimensional (1)H NMR spectra of the imino proton region of I in the presence of this series of compounds revealed that Blm and Zn- and CuBlm bind in fast exchange on the NMR time scale, while the Fe and Co complexes bind in slow exchange. Blm, ZnBlm, and Cu(II)Blm caused little perturbation of the UV circular dichroism spectrum of I or II. In contrast, Fe(III)Blm and HO(2)-Co(III)Blm induced hypochromic effects in the CD spectrum of I and altered the spectrum of II to a smaller extent. On the basis of these results, the DNA binding structures and properties of Blm A(2), ZnBlm A(2), and CuBlm A(2) differ substantially from those of Fe(III)Blm A(2) and HO(2)-Co(III)Blm A(2). PMID- 11412111 TI - Localization of a heparin binding site in the catalytic domain of factor XIa. AB - Inhibition of factor XIa by protease nexin II (K(i) approximately 450 pM) is potentiated by heparin (K(I) approximately 30 pM). The inhibition of the isolated catalytic domain of factor XIa demonstrates a similar potentiation by heparin (K(i) decreasing from 436 +/- 62 to 88 +/- 10 pM) and also binds to heparin on surface plasmon resonance (K(d) 11.2 +/- 3.2 nM vs K(d) 8.63 +/- 1.06 nM for factor XIa). The factor XIa catalytic domain contains a cysteine-constrained alpha-helix-containing loop: (527)CQKRYRGHKITHKMIC(542), identified as a heparin binding region in other coagulation proteins. Heparin-binding studies of coagulation proteases allowed a grouping of these proteins into three categories: group A (binding within a cysteine-constrained loop or a C-terminal heparin binding region), factors XIa, IXa, Xa, and thrombin; group B (binding by a different mechanism), factor XIIa and activated protein C; and group C (no binding), factor VIIa and kallikrein. Synthesized peptides representative of the factor XIa catalytic domain loop were used as competitors in factor XIa binding and inhibition studies. A native sequence peptide binds to heparin with a K(d) = 86 +/- 15 nM and competes with factor XIa in binding to heparin, K(i) = 241 +/- 37 nM. A peptide with alanine substitutions at (534)H, (535)K, (538)H, and (539)K binds and competes with factor XIa for heparin-binding in a manner nearly identical to that of the native peptide, whereas a scrambled peptide is approximately 10-fold less effective, and alanine substitutions at residues (529)K, (530)R, and (532)R result in loss of virtually all activity. We conclude that residues (529)K, (530)R, and (532)R comprise a high-affinity heparin-binding site in the factor XIa catalytic domain. PMID- 11412112 TI - Beta 93 modified hemoglobin: kinetic and conformational consequences. AB - The reactive sulfhydryl on Cys beta93 in human adult hemoglobin (HbA) has been the focus of much attention. It has purported functional roles such as a transporter of nitric oxide and a detoxifier of super oxide. In addition, it has a proposed role in the allosteric mechanism. The present study addresses the functional and conformational consequences of modifying the beta93 sulfhydryl using either maleimide or disulfide-based reactions. The geminate and bimolecular recombination of CO derivatives of several different beta93-modified Hbs in both solution and sol-gel matrixes provide a window into functional modifications associated with both the R and T states of these proteins. Nanosecond time resolved visible resonance Raman spectroscopy is used to probe conformational consequences associated with the proximal heme environment. The results show functional and conformational consequences that depend on the specific chemistry used to modify beta93. Maleimide-based modification show the most significant alterations of R-state properties including a consistent pattern of a reduced geminate yield and a loss of the favorable heme-proximal histidine interaction normally seen for liganded R-state HbA. A mechanism based on a displacement of the side chain of Tyr beta145 is explored as a basis for this effect as well as other situations where there is loss of the quaternary enhancement effect. The quaternary enhancement effect refers to the enhancement of ligand binding properties of the alphabeta dimers when they are associated into the R-state tetramer. PMID- 11412113 TI - In vitro reconstitution of recognition and activation complexes between interleukin-6 and gp130. AB - Gp130 is a shared signal-transducing receptor for a family of four-helix cytokines, of which interleukin-6 is a prototypic member. IL-6-type cytokines activate gp130 to elicit downstream intracellular JAK/STAT signaling cascades through formation of hetero-oligomeric receptor complexes. Interleukin-6 must first complex with its specific alpha-receptor (Ralpha) in order to bind and activate gp130. We have dissected the extracellular activation pathway of human gp130 by human IL-6 through reconstitution of soluble complexes representing intermediate and final states in the hierarchical assembly of the IL-6/IL 6Ralpha/gp130 signaling complex. To isolate these hetero-complexes, we have applied a protein engineering strategy of covalently linking IL-6 to its Ralpha, which results in a "hyperactive" single-chain complex (hyper-IL-6) which we express in both Escherichia coli and insect cells. We have determined that IL 6/IL-Ralpha and the cytokine-binding homology region (CHR) of gp130 (D2D3) form a stable trimolecular "recognition" complex (trimer) consisting of 1IL-6,1 IL 6Ralpha, and 1 gp130-CHR. Addition of the N-terminal (D1) Ig-like domain (IGD) of gp130 to the CHR results in a transition to a hexameric "activation" complex containing 2 IL-6, 2IL-6Ralpha, and 2 gp130. These results clearly demonstrate that the recognition and activation complexes are disparate hetero-oligomeric molecular species linked by the recruitment of the gp130 IGD by the unique site III epitope present on all gp130-class cytokines. The results of these studies are relevant to other members of the IL-6 family of gp130-cytokines and address a longstanding question concerning the respective roles of the gp130 CHR and IGD in assembly of the active signaling oligomer. PMID- 11412114 TI - Redox-dependent CO2 reduction activity of CO dehydrogenase from Rhodospirillum rubrum. AB - Carbon monoxide dehydrogenase (CODH) from Rhodospirillum rubrum catalyzes both the oxidation of CO and the reduction of CO(2). Studies of the redox dependence of CO(2) reduction by R. rubrum CODH show that (1) CODH is unable to catalyze CO(2) reduction at potentials greater than -300 mV; (2) the maximum activity is observed at potentials less than -480 mV; and (3) the midpoint potential (E(m)) of the transition from minimum to maximum CO(2) reduction activity occurs at approximately -339 mV. These results indicate that the C(red1) state of R. rubrum CODH (E(m) = -110 mV; g(zyx)() = 2.03, 1.88, 1.71) is not competent to reduce CO(2). Nernst analyses suggest that the reduction of CODH from the C(red1) state to the CO(2)-reducing form (C(unc), g(zyx)() = 2.04, 1.93, 1.89; E < approximately -300 mV) of the enzyme is a one-electron process. For the entire redox range, viologens stimulate CO(2) reduction by CODH more than 50-fold, and it is proposed that viologens accelerate the redox equilibration of redox buffers and [Fe(4)S(4)](B) during catalysis. PMID- 11412115 TI - Single turnover EPR studies of benzoyl-CoA reductase. AB - Benzoyl-CoA reductase (BCR) catalyzes the ATP-driven transport of two electrons from a reduced 2[4Fe-4S] ferredoxin to the aromatic ring of benzoyl-CoA. A mechanism involving radical species and very low potential electrons similar to the Birch reduction of aromatics has been suggested for this reaction. The redox centers of BCR have previously been identified, by EPR- and Mossbauer spectroscopy, to be three cysteine-ligated [4Fe-4S] clusters [Boll et al. (2000) J. Biol. Chem. 275, 31857-31868] with redox potentials more negative than -500 mV. In this work, the catalytic cycle of BCR was studied by freeze-quench experiments; the dithionite reduced enzyme was rapidly mixed with equimolar amounts of benzoyl-CoA and excess MgATP plus dithionite, and subjected to EPR spectroscopic analysis. The turnover period of the enzyme under the conditions used was 3 s. The total S = (1)/(2) spin concentration increased 3-fold very rapidly (within approximately 25 ms). In the course of a single turnover the extent of enzyme reduction decreased again, finally reaching the starting value. An increased magnetic interaction of [4Fe-4S] clusters and the rise of an S = (7)/(2) high-spin EPR signal occurred as second simultaneous and transient events (at approximately 200 ms). Previous work showed that binding of the nucleotide affects the magnetic interaction of [4Fe-4S] clusters, whereas hydrolysis of MgATP is required for the switch to high-spin EPR signals. Finally, two novel transient EPR signals with an isotropic line-shape developed maximally in the late phase of the catalytic cycle ( approximately 1-2 s). These signals differed from those of typical free radicals by shifted g values at g = 2.015 and g = 2.033 and by an unusually fast relaxation rate, suggesting an interaction of these paramagnetic species with [4Fe-4S](+1) clusters. On the basis of these results, we present a proposal for a catalytic cycle involving radical species. PMID- 11412116 TI - Putative helix F contributes to regioselectivity of hydroxylation in mitochondrial cytochrome P450 27A1. AB - On the basis of alignment with structurally characterized cytochromes P450 (P450s), we have identified the putative F and G helices of mitochondrial P450s 27A1 and 11A. We introduced substitutions at Phe-207, Ile-211, and Phe-215 within putative helix F and at Trp-235 and Tyr-238 within putative helix G in P450 27A1 and compared wild type and mutants with respect to catalytic activity, product pattern, substrate binding, formation of hydrogen peroxide, and interaction with redox partner. Results indicate that the mutated residues are important for delivery of the correctly oriented substrate to the P450 active site. The I211K and F215K mutations, for example, affected the regioselectivity of P450 27A1 dependent hydroxylation reactions and conferred the P450 capacity to cleave the C C bond of the substrate during the catalytic cycle. Studies of P450 11A1 indicate that Phe-202 has functions similar to those of its counterpart in P450 27A1 (Phe 215). We propose that putative helices F and G form the sides of the substrate access channel, thus providing the additional mechanism to control regioselectivity of hydroxylation in mitochondrial P450s. PMID- 11412117 TI - Complex O-acetylation in Legionella pneumophila serogroup 1 lipopolysaccharide. Evidence for two genes involved in 8-O-acetylation of legionaminic acid. AB - A putative gene encoding an O-acetyl transferase, lag-1, is involved in biosynthesis of the O-polysaccharide (polylegionaminic acid) in some Legionella pneumophila serogroup 1 strains. To study the effect of the presence and absence of the gene on the O-polysaccharide O-acetylation, lag-1 from strain Philadelphia 1 was expressed in trans in the naturally lag-1-negative OLDA strain RC1, and immunoblot analysis revealed that the lag-1-encoded O-acetyl transferase is active. O-Polysaccharides of different size were prepared from the lipopolysaccharides of wild-type and transformant strains by mild acid degradation followed by gel-permeation chromatography. Using NMR spectroscopy and MALDI-TOF mass spectrometry, it was found that O-acetylation of the first three legionaminic acid residues next to the core occurs in the short-chain O polysaccharide (<10 sugars) from both strains. Hence, there is another O-acetyl transferase encoded by a gene different from lag-1. In the longer-chain O polysaccharide, a legionaminic acid residue proximal to the core is N-methylated and could be further 8-O-acetylated in the lag-1-dependent manner. Only strains expressing a functional lag-1 gene were recognized in Western blot analysis by monoclonal antibody 3/1 requiring 8-O-acetylated polylegionaminic acid for binding. The highly O-acetylated outer core region of the lipopolysaccharide is involved in the epitope of another serogroup 1-specific monoclonal antibody termed LPS-1. The O-acetylation pattern of the L. pneumophila serogroup 1 core oligosaccharide was revised using MALDI-TOF mass spectrometry. lag-1-independent O-acetylation of the core and short-chain O-polysaccharide was found to be a common feature of L. pneumophila serogroup 1 strains. The biological importance of conserved lag-1-independent and variable lag-1-dependent O-acetylation is discussed. PMID- 11412118 TI - Effects of two photoreactive spermine analogues on peptide bond formation and their application for labeling proteins in Escherichia coli functional ribosomal complexes. AB - The effect of two photoreactive analogues of spermine, N(1)-azidobenzamidino- (ABA-) spermine and N(1)-azidonitrobenzoyl- (ANB-) spermine, on ribosomal functions was studied in a cell-free system derived from Escherichia coli. In the dark, both analogues stimulated the binding of AcPhe-tRNA to poly(U)-programmed ribosomes, enhanced the stability of the ternary complex AcPhe tRNA.poly(U).ribosome (complex C), and caused stimulatory and inhibitory effects on peptidyltransferase activity. ABA-spermine exhibited more pronounced effects than ANB-spermine. Each photoprobe was covalently attached after irradiation to both ribosomal subunits and also to free rRNA isolated from 70S ribosomes. Photolabeled complex C showed a reactivity toward puromycin, similar to that exhibited by complex C reacting reversibly with photoprobes free in solution. The distribution of the incorporated radioactivity among the ribosomal components was determined under two experimental conditions, one stimulating and the other inhibiting peptidyltransferase activity. Under both conditions, ABA-spermine was the strongest cross-linker. Upon stimulatory conditions, 14% of ABA [(14)C]spermine cross-linked to complex C was bound to the protein fraction. The proteins primarily labeled were identified as S3, S4, L2, L3, L6, L15, L17, and L18. Upon inhibitory conditions, a higher percent of the incorporated radioactivity was found in ribosomal proteins, while the pattern of protein labeling was characterized by a remarkable decrease of cross-linked proteins L2, L3, L6, L15, L17. and L18 and by an increase of cross-linked proteins S9, S18, L1, L16, L22, L23, and L27. On the basis of these results and literature data, the involvement of spermine in the conformation and important functions of ribosomes is discussed. PMID- 11412119 TI - Helicase assembly protein Gp59 of bacteriophage T4: fluorescence anisotropy and sedimentation studies of complexes formed with derivatives of Gp32, the phage ssDNA binding protein. AB - The gene 59 protein (gp59) of bacteriophage T4 performs a vital function in phage DNA replication by directing the assembly of gp41, the DNA helicase component of the T4 primosome, onto lagging strand ssDNA at nascent replication forks. The helicase assembly activity of gp59 is required for optimum efficiency of helicase acquisition by the replication fork during strand displacement DNA synthesis and is essential for helicase and primosome assembly during T4 recombination dependent DNA replication transactions. Of central importance is the ability of gp59 to load the gp41 helicase onto ssDNA previously coated with cooperatively bound molecules of gp32, the T4 ssDNA binding protein. Gp59 heteroassociations with ssDNA, gp32, and gp41 all appear to be essential for this loading reaction. Previous studies demonstrated that a tripartite complex containing gp59 and gp32 simultaneously cooccupying ssDNA is an essential intermediate in gp59-dependent helicase loading; however, the biochemical and structural parameters of gp59-gp32 complexes with or without ssDNA are currently unknown. To better understand gp59 gp32 interactions, we performed fluorescence anisotropy and analytical ultracentrifugation experiments employing native or rhodamine-labeled gp59 species in combination with altered forms of gp32, allowing us to determine their binding parameters, shape parameters, and other hydrodynamic properties. Two truncated forms of gp32 were used: gp32-B, which lacks the N-terminal B-domain required for cooperative binding to ssDNA and for stable self-association, and A domain fragment, which is the C-terminal peptide of gp32 lacking ssDNA binding ability. Results indicate that gp59 binds with high affinity to either gp32 derivative to form a 1:1 heterodimer. In both cases, heterodimer formation is accompanied by a conformational change in gp59 which correlates with decreased gp59-DNA binding affinity. Hydrodynamic modeling suggests an asymmetric prolate ellipsoid shape for gp59, consistent with its X-ray crystallographic structure, and this asymmetry appears to increase upon binding of gp32 derivatives. Implications of our findings for the structure and function of gp59 and gp59-gp32 complexes in T4 replication are discussed. PMID- 11412120 TI - Solid-state NMR investigation of the dynamics of the soluble and membrane-bound colicin Ia channel-forming domain. AB - Solid-state NMR spectroscopy was employed to study the molecular dynamics of the colicin Ia channel domain in the soluble and membrane-bound states. In the soluble state, the protein executes small-amplitude librations (with root-mean square angular fluctuations of 0-10 degrees ) in the backbone and larger amplitude motions (16-17 degrees ) in the side chains. Upon membrane binding, the motional amplitudes increase significantly for both the backbone (12-16 degrees ) and side chains (23-29 degrees ), as manifested by the reduction in the C-H and H H dipolar couplings and (15)N chemical shift anisotropy. These motions occur not only on the pico- to nanosecond time scales, but also on the microsecond time scale, as revealed by the (1)H rotating-frame spin-lattice relaxation times. Average motional correlation times of 0.8 and 1.2 micros were extracted for the soluble and membrane-bound states, respectively. In comparison, both forms of the colicin Ia channel domain are completely immobile on the millisecond scale. These results indicate that the colicin Ia channel domain has enhanced conformational mobility in the lipid bilayer compared to the soluble state. This membrane induced mobility increase is consistent with the loss of tertiary structure of the protein in the membrane, which was previously suggested by the extended helical array model [Zakharov et al. (1998) Proc. Natl. Acad. Sci. U.S.A. 95, 4282-4287]. An extended structure would also facilitate protein interactions with the mobile lipids and thus increase the protein internal motions. We speculate that the large mobility of the membrane-bound colicin Ia channel domain is a prerequisite for channel opening in the presence of a voltage gradient. PMID- 11412121 TI - Nitration and oxidation of a hydrophobic tyrosine probe by peroxynitrite in membranes: comparison with nitration and oxidation of tyrosine by peroxynitrite in aqueous solution. AB - It has been reported that peroxynitrite will initiate both oxidation and nitration of tyrosine, forming dityrosine and nitrotyrosine, respectively. We compared peroxynitrite-dependent oxidation and nitration of a hydrophobic tyrosine analogue in membranes and tyrosine in aqueous solution. Reactions were carried out in the presence of either bolus addition or slow infusion of peroxynitrite, and also using the simultaneous generation of superoxide and nitric oxide. Results indicate that the level of nitration of the hydrophobic tyrosyl probe located in a lipid bilayer was significantly greater than its level of oxidation to the corresponding dimer. During slow infusion of peroxynitrite, the level of nitration of the membrane-incorporated tyrosyl probe was greater than that of tyrosine in aqueous solution. Evidence for hydroxyl radical formation from decomposition of peroxynitrite in a dimethylformamide/water mixture was obtained by electron spin resonance spin trapping. Mechanisms for nitration of the tyrosyl probe in the membrane are discussed. We conclude that nitration but not oxidation of a tyrosyl probe by peroxynitrite is a predominant reaction in the membrane. Thus, the local environment of target tyrosine residues is an important factor governing its propensity to undergo nitration in the presence of peroxynitrite. This work provides a new perspective on selective nitration of membrane-incorporated tyrosine analogues. PMID- 11412122 TI - Roles of homooligomerization and membrane association in ATPase and proteolytic activities of FtsH in vitro. AB - Escherichia coli FtsH is a membrane-bound and ATP-dependent protease which degrades some soluble and integral membrane proteins. The N-terminal region of FtsH mediates membrane association as well as homooligomeric interaction of this enzyme. Previously, we studied in vivo functionality of FtsH derivatives, in which the N-terminal membrane region was either deleted (FtsH(DeltaTM)), replaced by a leucine zipper (Zip-FtsH(DeltaTM)), or replaced by a lactose permease transmembrane segment (LacY-FtsH). It was indicated that homooligomerization is required for the minimum proteolytic activity, whereas a transmembrane sequence is required for membrane protein degradation. Here we characterized these proteins in vitro. Although these mutant enzymes were very low in their activities, they were significantly stimulated by dimethyl sulfoxide, which enabled us to characterize their activities. LacY-FtsH degraded both soluble and membrane proteins, but Zip-FtsH(DeltaTM) only degraded soluble proteins. These proteins also exhibited significant ATPase activities. However, FtsH(DeltaTM) remained inactive both in ATPase and in protease activities even in the presence of dimethyl sulfoxide. The monomeric FtsH(DeltaTM) was able to bind ATP and a denatured protein. These results indicate that subunit association is important for the enzymatic catalysis by FtsH and that the additional presence of the transmembrane sequence is required for this enzyme to degrade a membrane protein even under detergent-solubilized conditions. PMID- 11412123 TI - The cysteine-rich amino-terminal domain of ZntA, a Pb(II)/Zn(II)/Cd(II) translocating ATPase from Escherichia coli, is not essential for its function. AB - Soft metal-translocating P1-type ATPases have a distinctive amino-terminal domain that contains one to six copies of the conserved metal-binding motif, GXXCXXC. ZntA from Escherichia coli, a Pb(II)-, Zn(II)-, and Cd(II)-transporting ATPase, has an approximately 120 residue amino-terminal domain with one copy of the GXXCXXC motif as well as four additional cysteine residues. The function of this domain was investigated by constructing a mutant of ZntA lacking the first approximately 100 residues. The mutant, DeltaN-ZntA, was able to confer resistance to Pb(II), Zn(II), and Cd(II) salts, in a manner similar to ZntA. The soft metal dependent ATP hydrolysis activity of purified DeltaN-ZntA was characterized. Purified DeltaN-ZntA and ZntA were both inactivated by oxidation. The K(m) for MgATP was unchanged for DeltaN-ZntA relative to ZntA. DeltaN-ZntA displayed the same metal ion specificity as ZntA. Thiolates increased the activities of both ZntA and DeltaN-ZntA. The V(max) values for DeltaN-ZntA were approximately 3-fold lower than for ZntA for all three metal ions. Thus, the amino-terminal domain is not essential for the function of ZntA or for conferring specificity toward particular soft metals. Its function may be to increase the overall catalytic rate by increasing the rate of metal ion binding to the transporter. Residues involved in the ATP-dependent soft metal ion-translocating mechanism as well as those responsible for recognition of specific metal ions must be part of the core structure of the P1-type ATPases. PMID- 11412124 TI - Enzyme-catalyzed condensation reaction in a mammalian alpha-amylase. High resolution structural analysis of an enzyme-inhibitor complex. AB - Mammalian alpha-amylases catalyze the hydrolysis of alpha-linked glucose polymers according to a complex processive mechanism. We have determined the X-ray structures of porcine pancreatic alpha-amylase complexes with the smallest molecule of the trestatin family (acarviosine-glucose) which inhibits porcine pancreatic alpha-amylase and yet is not hydrolyzed by the enzyme. A structure analysis at 1.38 A resolution of this complex allowed for a clear identification of a genuine single hexasaccharide species composed of two alpha-1,4-linked original molecules bound to the active site of the enzyme. The structural results supported by mass spectrometry experiments provide evidence for an enzymatically catalyzed condensation reaction in the crystal. PMID- 11412125 TI - Reconstructing the substrate for uracil DNA glycosylase: tracking the transmission of binding energy in catalysis. AB - The DNA repair enzyme uracil DNA glycosylase (UDG) is a powerful N-glycohydrolase that cleaves the glycosidic bond of deoxyuridine in DNA. We have investigated the role of substrate binding energy in catalysis by systematically dismantling the optimal substrate Ap(+1)UpA(-1)pA(-2) by replacing the nucleotides at the +1, -1, or -2 position with a tetrahydrofuran abasic site nucleotide (D), a 3 hydroxypropyl phosphodiester spacer (S), a phosphate monoester (p), or a hydroxyl group (h). Contrary to previous reports, the minimal substrate for UDG is 2' deoxyuridine (hUh). UDG has a significant catalytic efficiency (CE) for hUh of 4 x 10(7) M(-1) [CE = (k(cat)/K(m))(1/k(non)), where k(non) is the rate of the spontaneous hydrolysis reaction of hUh at 25 degrees C]. Addition of +1 and -1 phosphate monoanions to form pUp increases k(cat)/K(m) by 45-fold compared to that of hUh. The k(cat)/K(m) for pUp, but not pU or Up, is found to decrease by 20-fold over the pH range of 6-9 with a pK(a) of 7.1, which is identical to the pK(a) values for deprotonation of the +1 and -1 phosphate groups determined by the pH dependence of the (31)P NMR chemical shifts. This pH dependence indicates that binding of the pUp tetraanion is disfavored, possibly due to unfavorable desolvation or electrostatic properties of the highly charged +1 and -1 phosphate groups. Addition of flexible hydroxypropyl groups to the +1 and -1 positions to make SpUpS increases k(cat)/K(m) by more than 10(5)-fold compared to that of hUh, which is a 20-fold greater effect than observed with rigid D substituents in these positions (i.e., DpUpD). The -2 phosphoester or nucleotide is found to increase the reactivity of trimer substrates with rigid furanose rings or nucleotides in the +1 and -1 positions by 1300-270000-fold (i.e., DpUpD --> DpUpDpA or ApUpA --> ApUpApA). In contrast, the -2 nucleotide provides only an 8 fold rate enhancement when appended to the substrate containing the more flexible +1 and -1 S substituents (SpUpS --> SpUpSpA). These context-dependent effects of a -2 nucleotide are interpreted in terms of a mechanism in which the binding energy of this "handle" is used drive the rigid +1 and -1 A or D substituents into their binding pockets, resulting in a net catalytic benefit of -4.3 to -7.5 kcal/mol. Taken together, these results systematically track how UDG uses distant site binding interactions to produce an overall four billion-fold increase in CE compared to that of the minimal substrate hUh. PMID- 11412126 TI - Role of aromatic stacking interactions in the modulation of the two-electron reduction potentials of flavin and substrate/product in Megasphaera elsdenii short-chain acyl-coenzyme A dehydrogenase. AB - The effects of aromatic stacking interactions on the stabilization of reduced flavin adenine dinucleotide (FAD) and substrate/product have been investigated in short-chain acyl-coenzyme A dehydrogenase (SCAD) from Megasphaera elsdenii. Mutations were made at the aromatic residues Phe160 and Tyr366, which flank either face of the noncovalently bound flavin cofactor. The electrochemical properties of the mutants were then measured in the presence and absence of a butyryl-CoA/crotonyl-CoA mixture. Results from these redox studies suggest that the phenylalanine and tyrosine both engage in favorable pi-sigma interactions with the isoalloxazine ring of the flavin to help stabilize formation of the anionic flavin hydroquinone. Disruption of these interactions by replacing either residue with a leucine (F160L and Y366L) causes the midpoint potential for the oxidized/hydroquinone couple (E(ox/hq)) to shift negative by 44-54 mV. The E(ox/hq) value was also found to decrease when aromatic residues containing electron-donating heteroatoms were introduced at the 160 position. Potential shifts of -32 and -43 mV for the F160Y and F160W mutants, respectively, are attributed to increased pi-pi repulsive interactions between the ring systems. This study also provides evidence for thermodynamic regulation of the substrate/product couple in the active site of SCAD. Binding to the wild-type enzyme caused the midpoint potential for the butyryl-CoA/crotonyl-CoA couple (E(BCoA/CCoA)) to shift 14 mV negative, stabilizing the oxidized product. Formation of product was found to be even more favorable in complexes with the F160Y and F160W mutants, suggesting that the electrostatic environment around the flavin plays a role in substrate/product activation. PMID- 11412127 TI - Interaction of the catch-loop tyrosine beta Y317 with the metal at catalytic site 3 of Chlamydomonas chloroplast F1-ATPase. AB - Site-directed mutants Y317C, Y317E, Y317F, Y317G, and Y317K were made to the catch-loop tyrosine on the beta subunit of the chloroplast F(1)-ATPase in Chlamydomonas. EPR spectra of VO(2+)-ATP bound to site 3 of CF(1) from wild type and mutants were obtained. Every mutant changed the (51)V hyperfine parameters of the VO(2+) bound at this site in the catalytically active conformation of the enzyme but had no effect on these parameters in the form that predominates when the enzyme activity is latent. These results indicate that this residue is a ligand to the metal of the Mg(2+)-nucleotide complex that binds to the empty catalytic site. The mutations also decreased the k(cat) of the ATPase activity to a much greater extent than k(cat)/K(M). Thus, these mutations limit the rate of product (Mg(2+)-ADP and phosphate) release in the ATPase direction or, conversely, the initial binding of substrates in the ATP synthesis direction. On the basis of these observations, coordination of betaY317 by Mg(2+)-ADP that binds to the empty catalytic site provides a means by which substrate binding could trigger gamma subunit rotation and consequent conformation changes of beta subunits during ATP synthesis. PMID- 11412128 TI - Slow-binding inhibition of human prostaglandin endoperoxide synthase-2 with darbufelone, an isoform-selective antiinflammatory di-tert-butyl phenol. AB - The antiinflammatory agent darbufelone, ((Z)-5-[[3,5-bis(1,1-dimethylethyl)-4 hydroxyphenyl] methylene]-2-imino-4-thiazolidinone, methanesulfonate salt), was discovered as a dual inhibitor of cellular prostaglandin and leukotriene production. To study the mechanism of action of this drug, we expressed human prostaglandin endoperoxide synthase-1 (PGHS-1) and PGHS-2 and purified the recombinant enzymes using buffers that contain octylglucoside. In cyclooxygenase assays following a 15-min incubation of enzyme with inhibitor, darbufelone potently inhibits PGHS-2 (IC(50) = 0.19 microM) but is much less potent with PGHS 1 (IC(50) = 20 microM). Interestingly, when the assay buffer contains traces of Tween 20 (0.0001%), darbufelone appears inactive with PGHS-2 due to a detergent interaction that is detectable by absorption spectroscopy. We therefore used octylglucoside, which does not affect darbufelone in this way, in place of Tween 20 in our PGHS buffers. Inhibition of PGHS-2 with darbufelone is time dependent: with no preincubation, darbufelone is a weak inhibitor (IC(50) = 14 microM), but after a 30-min incubation it is 20-fold more potent. Plots of PGHS-2 activity vs preincubation time at various darbufelone concentrations reach a plateau. This finding is inconsistent with irreversible or one-step slow-binding inhibition. A two-step slow-binding inhibition model is proposed in which the E.I complex (K(i) = 6.2 +/- 1.9 to 14 +/- 1 microM) slowly transforms (k(5) = 0.015-0.030 s(-)(1)) to a tightly bound E.I form with K(i) = 0.63 +/- 0.07 microM and k(6) = 0.0034 s( )(1). In steady-state kinetics inhibition experiments performed with no preincubation, we find that darbufelone is a noncompetitive inhibitor of PGHS-2 (K(i) = 10 +/- 5 microM). Darbufelone quenches the fluorescence of PGHS-2 at 325 nm (lambda(ex) = 280 nm) with K(d) = 0.98 +/- 0.03 microM. The PGHS substrate, arachidonate, and various cyclooxygenase inhibitors do not alter this binding affinity of darbufelone but a structural analogue of darbufelone competes directly for binding to PGHS-2. Di-tert-butyl phenols such as darbufelone may inhibit PGHS-2 by exploiting a previously unrecognized binding site on the enzyme. PMID- 11412129 TI - Thermodynamic characterization of the human acidic fibroblast growth factor: evidence for cold denaturation. AB - The thermodynamic parameters characterizing the conformational stability of the human acidic fibroblast growth factor (hFGF-1) have been determined by isothermal urea denaturation and thermal denaturation at fixed concentrations of urea using fluorescence and far-UV CD circular dichroism (CD) spectroscopy. The equilibrium unfolding transitions at pH 7.0 are adequately described by a two-state (native < -> unfolded state) mechanism. The stability of the protein is pH-dependent, and the protein unfolds completely below pH 3.0 (at 25 degrees C). hFGF-1 is shown to undergo a two-state transition only in a narrow pH range (pH 7.0-8.0). Under acidic (pH <6.0) and basic (pH >8.0) conditions, hFGF-1 is found to unfold noncooperatively, involving the accumulation of intermediates. The average temperature of maximum stability is determined to be 295.2 K. The heat capacity change (DeltaC(p)()) for the unfolding of hFGF-1 is estimated to be 2.1 +/- 0.5 kcal.mol(-1).K(-1). Temperature denaturation experiments in the absence and presence of urea show that hFGF-1 has a tendency to undergo cold denaturation. Two-dimensional (1)H-(15)N HSQC spectra of hFGF-1 acquired at subzero temperatures clearly show that hFGF-1 unfolds under low-temperature conditions. The significance of the noncooperative unfolding under acidic conditions and the cold denaturation process observed in hFGF-1 are discussed in detail. PMID- 11412130 TI - Conformational changes of an exchangeable apolipoprotein, apolipophorin III from Locusta migratoria, at low pH: correlation with lipid binding. AB - Locusta migratoria apolipophorin III (apoLp-III) is a helix bundle exchangeable apolipoprotein that reversibly binds to lipoprotein surfaces. Structural reorganization of its five amphipathic alpha-helices enables the transition from the lipid-free to lipid-bound state. ApoLp-III-induced transformation of dimyristoylphosphatidylcholine (DMPC) bilayer vesicles into smaller discoidal complexes is enhanced as a function of decreasing pH, with maximal transformation occurring at pH 3.5. Over the entire pH range studied, apoLp-III retains nearly all of its secondary structure content. Whereas no changes in fluorescence emission maximum of the two Trp residues in apoLp-III were observed in the pH range from 7.0 to 4.0, a further decrease in pH resulted in a strong red shift. Near-UV circular dichroism spectra of apoLp-III showed well-defined extrema (at 286 and 292 nm) between pH 7.0 and pH 4.0, which were attributed to Trp115. Below pH 4.0, these extrema collapsed, indicating a less rigid environment for Trp115. Similarly, the fluorescence intensity of 8-anilinonaphthalene-1-sulfonate in the presence of apoLp-III increased 4-fold below pH 4.0, indicating exposure of hydrophobic sites in the protein in this pH range. Taken together, the data suggest two conformational states of the protein. In the first state between pH 7.0 and pH 4.0, apoLp-III retains a nativelike helix bundle structure. The second state, found between pH 3.0 and pH 4.0, is reminiscent of a molten globule, wherein tertiary structure contacts are disrupted without a significant loss of secondary structure content. In both states DMPC vesicle transformation is enhanced by lowering the solution pH, reaching an optimum in the second state. The correlation between tertiary structure and lipid binding activity suggests that helix bundle organization is a determinant of apoLp-III lipid binding activity. PMID- 11412131 TI - Skeletal muscle diseases--common denominators. Proceedings of the 5th International Symposium. Sweden, 17-21 May 2000. PMID- 11412132 TI - The physiology of inflammatory myopathies: an overview. AB - The idiopathic inflammatory myopathies (IIMs) encompass a group of muscle disorders of unknown origin and pathogenesis characterized by symmetrical, proximal muscle weakness and by inflammatory infiltrates in muscle tissue. The mechanisms behind the loss of muscle function are largely unknown. It is often anticipated that the muscle weakness is caused by the inflammatory cells. However, inflammatory infiltrates are not always present in the muscle tissue and the infiltrates sometimes have a patchy distribution, which makes it difficult to explain the generalized muscle weakness merely by infiltration of inflammatory cells. We investigated patients at different stages of myositis: early myositis without detectable inflammatory infiltrates, active myositis with pronounced inflammatory infiltrates and chronic myositis with persisting muscle weakness but without detectable inflammatory cells in muscle tissues. In these studies, a better correlation was observed between the clinical symptoms and involvement of the capillaries with expression of the cytokine interleukin (IL)-1alpha and by the presence of major histocompatibility complex (MHC) class I expression on muscle fibres. Whether these molecules could affect muscle function is not known. Using phosphorus P-31 magnetic resonance spectroscopy decreased values of adenosine triphosphate (ATP) and phosphocreatine (PCr) levels were observed at rest. These metabolic abnormalities were further accentuated by exercise and increased PCr levels correlated with improved clinical status. The underlying mechanisms responsible for these biochemical abnormalities have not been defined but could be related to a disturbed tissue oxygenation. PMID- 11412133 TI - Immunological capabilities of skeletal muscle cells. AB - Muscle is the target of immunological injury in several muscle diseases. It is important therefore to understand the immunological capabilities of muscle cells themselves. Although it is conventional to discuss the effects of the immune system on other cells, tissues or organs, the system's boundaries cannot be sharply drawn, and in an increasing number of ways, the immunological capabilities of non-immune tissues are recognized as determining the course of immune-inflammatory processes. Muscle cells have an inherent ability to express and respond to a variety of immunologically relevant surface molecules, cytokines, and chemokines under inflammatory conditions. The ability of muscle cells to process and present antigens to the immune cells is currently debated; thus, this review is aimed at examining the immunological capabilities of skeletal muscle cells in vitro and in vivo. PMID- 11412134 TI - Cytokines and oxidative signalling in skeletal muscle. AB - A growing body of literature indicates that cytokines regulate skeletal muscle function, including gene expression and adaptive responses. Tumour necrosis factor-alpha (TNF-alpha) is the cytokine most prominently linked to muscle pathophysiology and, therefore, has been studied most extensively in muscle-based systems. TNF-alpha is associated with muscle catabolism and loss of muscle function in human diseases that range from cancer to heart failure, from arthritis to AIDS. Recent advances have established that TNF-alpha causes muscle weakness via at least two mechanisms, accelerated protein loss and contractile dysfunction. Protein loss is a chronic response that occurs over days to weeks. Changes in gene expression required for TNF-alpha induced catabolism are regulated by the transcription factor nuclear factor-kappaB which is essential for the net loss of muscle protein caused by chronic TNF-alpha exposure. Contractile dysfunction is an acute response to TNF-alpha stimulation, developing over hours and resulting in decreased force production. Both actions of TNF-alpha involve a rapid rise in endogenous oxidants as an essential step in post-receptor signal transduction. These oxidants appear to include reactive oxygen species derived from mitochondrial electron transport. Such information provides insight into the cellular and molecular mechanisms of TNF-alpha action in skeletal muscle and establishes a scientific basis for continued research into cytokine signalling. PMID- 11412135 TI - Exercise-induced muscle damage and inflammation: fact or fiction? AB - Physical exercise is necessary for maintaining normal function of skeletal muscle. The mechanisms governing normal muscle function and maintenance are vastly unknown but synergistic function of hormones, neurosignalling, growth factors, cytokines and other factors, is undoubtedly important. Because of the complex interaction among these systems the lack of complete understanding of muscle function is not surprising. The purpose of exercise-induced changes in muscle cell function is to adapt the tissue to a demand of increased physical work capacity. Some of the approaches used to investigate changes in skeletal muscle cell function are exercise and electrical stimulation in animals and human models and isolated animal muscle. From these models, it has been concluded that during physical exercise, in an intensity and duration dependent manner, skeletal muscle is damaged and subsequently inflamed. The purpose of the inflammation would be to repair the exercise-induced damage. Because of the design and methods used in a majority of these studies, concerns must be raised, and the question asked whether the paradigm of exercise-induced muscle inflammation in fact is fiction. In a majority of conducted studies, a non-exercising control group is lacking and because of the invasive nature of the sampling methods used to study inflammation it does not appear impossible that observed inflammatory events in human skeletal muscle after physical exercise are methodological artefacts. PMID- 11412136 TI - The pathophysiology of type 2 diabetes mellitus: an overview. AB - Type 2 diabetes mellitus is a heterogeneous disorder characterized by chronic hyperglycaemia. The aetiological heterogeneity is suggested by genetic inheritance and its interplay with environmental factors. Impaired insulin secretion and decreased insulin sensitivity are the main pathophysiological features, responsible for development of hyperglycaemia in type 2 diabetes. However, the genetic basis of these defects has been demonstrated only in small subgroups of patients. Whether impaired secretion or action of insulin is the primary defect in the majority of patients is not known, although it is generally agreed that defective insulin release is a requirement for the disease to develop. PMID- 11412137 TI - Intracellular mechanisms underlying increases in glucose uptake in response to insulin or exercise in skeletal muscle. AB - This review will provide insight on potential intracellular signalling mechanisms by which insulin and exercise/contraction increases glucose metabolism and gene expression. Glucose transport, the rate limiting step in glucose metabolism, is mediated by glucose transporter 4 (GLUT4) and can be activated in skeletal muscle by two separate and distinct signalling pathways; one stimulated by insulin and the second by muscle contractions. Impaired insulin action on whole body glucose uptake is a hallmark feature of type II (non-insulin-dependent) diabetes mellitus. Defects in insulin signal transduction through the insulin-receptor substrate-1/phosphatidylinositol 3-kinase pathway are associated with reduced insulin-stimulated glucose transporter 4 translocation and glucose transport activity in skeletal muscle from type II diabetic patients. Studies performed using glucose transporter 4-null mice show that this glucose transporter isoform plays a major role in mediating exercise-stimulated glucose uptake in skeletal muscle. Level of physical exercise has been linked to improved glucose homeostasis and enhanced insulin sensitivity. Exercise training leads to alterations in expression and activity of key proteins involved in insulin signal transduction. These changes may be related to increased signal transduction through the mitogen-activated protein kinase (MAPK) signalling cascades. Because MAPK is associated with increased transcriptional activity, these signalling cascades are candidates for these exercise-induced changes in protein expression. Understanding the molecular mechanism for the activation of signal transduction pathways will provide a link for defining new strategies to enhance glucose metabolism and improve health in the general population. PMID- 11412138 TI - The role of Ca2+ and calmodulin in insulin signalling in mammalian skeletal muscle. AB - The role of Ca2+ in mediating effects of insulin on skeletal muscle has been widely debated. It is believed that in skeletal muscle Ca2+ has a permissive role, necessary but not of prime importance in mediating the stimulatory actions of insulin. In this review, we present evidence that insulin causes a localized increase in the concentration of Ca2+. Specifically, insulin induces a rise in near-membrane Ca2+ but not the bulk Ca2+ in the myoplasm. The rise in near membrane Ca2+ is because of an influx through channels that can be blocked by L type Ca2+ channel inhibitors. Calcium appears to exert some of its subsequent effects via calmodulin-dependent processes as calmodulin inhibitors block the translocation of glucose transporters and other enzymes as well as the insulin stimulated increase in glucose transport. PMID- 11412139 TI - Relationship between muscle fibre composition, glucose transporter protein 4 and exercise training: possible consequences in non-insulin-dependent diabetes mellitus. AB - Skeletal muscle is composed of different fibre types, which differ in contractile as well as in metabolic properties. The myosin molecule, which exists in several different isoforms, is of major importance in determining the contractile properties of the muscle cell. The plasticity of skeletal muscle is reflected in this tissue's adaptability to changes in the functional demand. In both rats and humans, a decrease in activity level will in most cases change the muscle fibre composition towards faster myosin isoforms and an increase in activity level (such as seen with exercise training) will induce an increase in slower myosin isoforms. The glucose transporter protein 4 (GLUT4), which is the major insulin regulatable glucose transporter in mammalian skeletal muscle, is found in larger amounts in slow muscle fibres compared with fast muscle fibres. An increase in activity level will increase the GLUT4 protein expression and a decrease in activity level will in most cases decrease GLUT4. Thus, there seems to be some kind of relationship between the muscle fibre type and GLUT4. However, the main factor regulating both the GLUT4 protein expression and the muscle fibre composition seems to be the activity level of the muscle fibre. Patients suffering from non-insulin-dependent diabetes mellitus (NIDDM) are insulin resistant in their skeletal muscles but are generally normal when it comes to skeletal muscle fibre composition and the GLUT4 protein expression. There is good evidence that exercise training beneficially impacts on insulin sensitivity in healthy individuals and in patients with type II diabetes. An increase in the GLUT4 protein expression in skeletal muscle may at least partly explain this effect of training. PMID- 11412140 TI - Skeletal muscle disorders in heart failure. AB - Heart failure is associated with reduction of exercise capacity that cannot be solely ascribed to reduced maximal oxygen uptake (VdotO2max). Therefore, research has focused on changes in skeletal muscle morphology, metabolism and function. Factors that can cause such changes in skeletal muscle comprise inactivity, malnutrition, constant or repeated episodes of inadequate oxygen delivery and prolonged exposure to altered neurohumoural stimuli. Most of these factors are not specific for the heart failure condition. On the other hand, heart failure is more than one clinical condition. Congestive heart failure (CHF) develops gradually as a result of deteriorating contractility of the viable myocardium, myocardial failure. Is it possible that development of this contractile deficit in the myocardium is paralleled by a corresponding contractile deficit of the skeletal muscles? This question cannot be answered today. Both patient studies and experimental studies support that there is a switch to a faster muscle phenotype and energy metabolism balance is more anaerobic. The muscle atrophy seen in many patients is not so evident in experimental studies. Few investigators have studied contractile function. Both fast twitch and slow twitch muscles seem to become slower, not faster as might be expected, and this is possibly linked to slower intracellular Ca2+ cycling. The neurohumoural stimuli that can cause this change are not known, but recently it has been reported that several cytokines are increased in CHF patients. Thus, the changes seen in skeletal muscles during CHF are partly secondary to inactivity, but the possibility remains that the contractility is altered because of intracellular changes of Ca2+ metabolism that are also seen in the myocardium. PMID- 11412141 TI - Skeletal muscle training in chronic heart failure. AB - Patients with heart failure are limited in their ability to tolerate exercise. Recent research has suggested that this limitation cannot be entirely attributed to cardiac or lung impairment but rather that changes in peripheral muscles may play an important role. There are objective similarities between heart failure and muscular deconditioning. Deficiencies in peripheral blood flow and skeletal muscle function, morphology, metabolism and function are present in both conditions. Moreover, an exaggerated activity of the receptors sensitive to exercise-derived metabolic signals (muscle ergoreceptors and peripheral and central chemoreceptors) leads to early and profound exercise-induced fatigue and dyspnoea. These muscle afferents contribute to the ventilatory, haemodynamic and autonomic responses to exercise both in physiological and pathological conditions, including chronic heart failure. Against this background, a skeletal muscle origin of symptoms in heart failure has been proposed. The protective effects of physical training have been described in many recent studies: training improves ventilatory control, skeletal muscle metabolism and autonomic nervous system activity. The exercise training appears to induce its beneficial effects on skeletal muscle both directly (on muscle function, histological and biochemical features) and indirectly (by reducing the activation of the muscle afferents). The metabolic mediators of these muscle afferents may become a potential target in the future therapy of heart failure symptoms. PMID- 11412142 TI - Apoptosis and changes in contractile protein pattern in the skeletal muscle in heart failure. AB - Chronic heart failure is characterized as a clinical disorder by exercise intolerance. There are two factors that are independently responsible for the reduced exercise capacity: (a) a shift from myosin heavy chain 1 (MHC1) to MHC2a and MHC2b and (b) muscle atrophy. We have demonstrated, both in experimental models of heart failure and in man, that the more severe the heart failure, the greater the magnitude of skeletal muscle apoptosis. In the monocrotaline treated rat, that develops a severe right-sided heart failure, the increased number of apoptotic nuclei was paralleled by increasing levels of circulating TNFalpha. In agreement with some recent observations showing that sphingolipids can mediate programmed cell death, we found that in animals with heart failure and high number of apoptotic nuclei, circulating levels of sphingosine were significantly increased. In a study conducted in patients with heart failure we found a correlation between exercise capacity limitation and skeletal myocytes apoptosis. There was also a correlation between degree of muscle atrophy and magnitude of apoptosis. The shift in MHCs, although with a different mechanism, is also responsible for the reduced exercise capacity in these patients. In fact there is a strong correlation between indices of severity of CHF and MHC composition. Muscle fatigue, appears earlier in patients that have a greater skeletal muscle expression of 'fast' MHCs. We have also demonstrated that MHCs shift and apoptosis can be prevented by using angiotensin II converting enzyme inhibitors and angiotensin II receptor blockers. PMID- 11412143 TI - Eccentric muscle damage: mechanisms of early reduction of force. AB - Pain and weakness are prominent symptoms which occur after a delay in muscles which have been stretched during contraction (eccentric contraction). These symptoms are particularly severe when the exercise is unaccustomed and when the stretch occurs in muscles on the descending limb of the force-length relation, i.e. at long muscle lengths. It is known that sarcomeres are potentially unstable on the descending limb and it has been proposed by Morgan that uncontrolled elongation of some sarcomeres occurs during eccentric contractions on the descending limb. In this article, the evidence that this mechanism leads to the reduced force is considered. If overextended sarcomeres persist after the eccentric exercise it will cause a shift in the peak of the force-length curve. There is also evidence that in some types of muscle, excitation-contraction coupling is impaired and contributes to the muscle weakness. Cytoskeletal proteins stabilize the sarcomeric structure and may be injured either by the overextended sarcomeres or by activation of proteases. The potential of these mechanisms to contribute to the effects of muscle training and to the symptoms of muscle disease, such as muscular dystrophy, is considered. PMID- 11412144 TI - Eccentric exercise-induced injuries to contractile and cytoskeletal muscle fibre components. AB - Exercise involving lengthening of an activated muscle can cause injury. Recent reports documented the mechanics of exercise-induced muscle injury as well as physiological and cellular events and manifestations of injury. Loss of the cytoskeletal protein desmin and loss of cellular integrity as evidenced by sarcolemmal damage occur early during heavy eccentric exercise. These studies indicate that the earliest events in muscle injury are mechanical in nature, while later events indicate that it may be more appropriate to conclude that intense exercise initiates a muscle remodeling process. We conclude that muscle injury after eccentric exercise is differently severe in muscles with different architecture, is fibre type-specific, primarily because of fibre strain in the acute phase, and is exacerbated by inflammation after the initial injury. PMID- 11412145 TI - Excitation-induced Ca2+ influx and skeletal muscle cell damage. AB - Excessive exercise may lead to skeletal muscle cell damage with degradation of cellular components and leakage of intracellular enzymes. Calcium has repeatedly been proposed to be involved in these processes. Studies have shown that the resting level of cytoplasmic Ca2+ increases up to threefold during long-term low frequency stimulation. We have shown that electrical stimulation produces a marked increase in Ca2+ uptake and Ca2+ content in rat skeletal muscle, both in vivo and in vitro. Continuous stimulation for 240 min at 1 Hz results in an increased release (18-fold) of lactate dehydrogenase (LDH) from extensor digitorum longus (EDL) muscle. This was associated with an increased total Ca2+ content (185%), was augmented at high [Ca2+]o and suppressed at low [Ca2+]o. The release of LDH may reflect partial loss of sarcolemmal integrity as a result of degradation of membrane components by Ca2+-activated enzymes (e.g. calpain or phospholipase A2). After cessation of stimulation the increased release of LDH continues for at least 120 min. This is associated with an up to sevenfold increase in 45Ca uptake. The increased permeability to Ca2+ may further activate calpain and phospholipase A2 and accelerate the loss of membrane integrity. Stimulation-induced uptake of Ca2+ and release of LDH is most pronounced in EDL (mainly composed of fast-twitch fibres at variance with soleus which is mainly composed of slow-twitch fibres). This may account for the observation that prolonged exercise leads to preferential damage to fast-twitch fibres. We hypothesize that excessive exercise may lead to an intracellular accumulation of Ca2+ and increased cytoplasmic Ca2+ causing activation of self-accelerating degradative pathways leading to muscle damage. PMID- 11412146 TI - Muscular effects in late polio. AB - New or increased muscular weakness, fatigue and muscle and joint pain with neuropathic electromyography (EMG) changes in a person with a confirmed history of polio constitute the cardinal symptoms of the post-polio syndrome. Unusual tiredness or fatigue is a common complaint in late polio subjects as is intolerance to cold. Fatigue in polio subjects can have several explanations: emotional fatigue, central nervous system fatigue, 'general' fatigue and/or neuromuscular fatigue. Some studies indicate central fatigue, but it is unclear how often and to which degree there will be a central muscular fatigue. Polio patients are known to be deconditioned (reduced function because of low activity level), and aerobic power is reduced. Defects in the neuromuscular transmission may be present but are not seen in all post-polio subjects with reduction in force and increased fatigability. The fatigue experienced by late polio patients is most likely an augmented peripheral muscle fatigue. Possible explanations may be an imperfection in the sarcoplasmatic reticulum with altered calcium release mechanisms (activation) or in sliding filament function (contractile properties). This may be a secondary effect to the enlarged muscle fibres. However, the prolonged subjective feeling of fatigue reported despite unchanged maximal voluntary contraction (MVC) remains unexplained. PMID- 11412147 TI - Desmin-related myopathies in mice and man. AB - Desmin, the main intermediate filament (IF) protein in skeletal and heart muscle cells, is of great importance as a part of the cytoskeleton. The IFs surround and interlink myofibrils, and connect the peripheral myofibrils with the sarcolemma. In myotendinous junctions and neuromuscular junctions of skeletal muscle fibres, desmin is enriched. In the heart, desmin is increased at intercalated discs, the attachment between cardiomyocytes, and it is the main component in Purkinje fibres of the conduction system. Desmin is the first muscle-specific protein to appear during myogenesis. Nevertheless, lack of desmin, as shown from experiments with desmin knockout (K/O) mice, does not influence myogenesis or myofibrillogenesis. However, the desmin knock-out mice postnatally develop a cardiomyopathy and a muscle dystrophy in highly used skeletal muscles. In other skeletal muscles the organization of myofibrils is remarkably unaffected. Thus, the main consequence of the lack of desmin is that the muscle fibres become more susceptible to damage. The loss of membrane integrity leads to a dystrophic process, with degeneration and fibrosis. In the heart cardiac failure develops, whereas in affected skeletal muscles regenerative attempts are seen. In humans, accumulations of desmin have been a hallmark for presumptive desmin myopathies. Recent investigations have shown that some families with such a myopathy have a defect in the gene coding for alphaB-crystallin, whereas others have mutations in the desmin gene. Typical features of these patients are cardiac affections and muscle weakness. Thus, mutations in the desmin gene is pathogenic for a distinct type of muscle disorder. PMID- 11412148 TI - Harnessing the potential of dystrophin-related proteins for ameliorating Duchenne's muscular dystrophy. AB - Duchenne's muscular dystrophy (DMD) is a fatal disease caused by mutations in the DMD gene that lead to quantitative and qualitative disturbances in dystrophin expression. Dystrophin is a member of the spectrin superfamily of proteins. Dystrophin itself is closely related to three proteins that constitute a family of dystrophin-related proteins (DRPs): the chromosome 6-encoded DRP or utrophin, the chromosome-X encoded, DRP2 and the chromosome-18 encoded, dystrobrevin. These proteins share sequence similarity and functional motifs with dystrophin. Current attempts at somatic gene therapy of DMD face numerous technical problems. An alternative strategy for DMD therapy, that circumvents many of these problems, has arisen from the demonstration that the DRP utrophin can functionally substitute for the missing dystrophin and its overexpression can rescue dystrophin-deficient muscle. Currently, a promising avenue of research consists of identifying molecules that would increase the expression of utrophin and the delivery of these molecules to dystrophin-deficient tissues as a means of DMD therapy. In this review, we will focus on DRPs from the perspective of strategies and issues related to upregulating utrophin expression for DMD therapy. Additionally, we will address the techniques used for anatomical, biochemical and physiological evaluation of the potential benefits of this and other forms of DMD therapy in dystrophin-deficient animal models. PMID- 11412149 TI - Muscle training in muscular dystrophies. AB - There has been a debate for many years on whether muscular training is beneficial or harmful for patients with myopathic disorders and the role of exercise training in the management of these patients is still controversial. Much of this confusion is because of the lack of well-designed controlled training studies on this heterogenic group of disorders. Because effective therapies are still lacking, the patients have to rely on symptomatic treatment in which continuous physiotherapy plays an important role. There is thus still a need for studies evaluating the short- and long-term effects of muscular training in different types of myopathic disorders. We need to elucidate whether muscular training can increase strength and resistance to fatigue, but most importantly, we need to clarify whether training can improve specific functional abilities of the patient with myopathy. Future studies should give us specific information on what type of training, endurance or strength training, is to be preferred for different myopathies. The effect of strength training in one type of muscle disorder is not directly applicable to another, but is largely dependent on the underlying biological defect. From the studies published so far, high-resistance strength training at submaximal and possibly also at near-maximal levels seem beneficial, at least in the short perspective for slowly progressive myopathic disorders. However, the long-term effects of such training have not been systematically studied. In rapidly progressive myopathies, which are caused by deficient structural proteins such as in Duchenne's muscular dystrophy, the use of high resistance training is far more controversial and questionable. If exercise regimens are to be used, they should preferably commence in the early stages of the disease, at which time there is still a substantial amount of trainable muscle fibres. PMID- 11412150 TI - Malignant hyperthermia and excitation-contraction coupling. AB - Malignant hyperthermia (MH) is a state of elevated skeletal muscle metabolism that may occur during general anaesthesia in genetically pre-disposed individuals. Malignant hyperthermia results from altered control of sarcoplasmic reticulum (SR) Ca2+ release. Mutations have been identified in MH-susceptible (MHS) individuals in two key proteins of excitation-contraction (EC) coupling, the Ca2+ release channel of the SR, ryanodine receptor type 1 (RyR1) and the alpha1-subunit of the dihydropyridine receptor (DHPR, L-type Ca2+ channel). During EC coupling, the DHPR senses the plasma membrane depolarization and transmits the information to the ryanodine receptor (RyR). As a consequence, Ca2+ is released from the terminal cisternae of the SR. One of the human MH-mutations of RyR1 (Arg614Cys) is also found at the homologous location in the RyR of swine (Arg615Cys). This animal model permits the investigation of physiological consequences of the homozygously expressed mutant release channel. Of particular interest is the question of whether voltage-controlled release of Ca2+ is altered by MH-mutations in the absence of MH-triggering substances. This question has recently been addressed in this laboratory by studying Ca2+ release under voltage clamp conditions in both isolated human skeletal muscle fibres and porcine myotubes. PMID- 11412151 TI - Autoimmunity against the ryanodine receptor in myasthenia gravis. AB - Some myasthenia gravis (MG) patients have antibodies against skeletal muscle antigens in addition to the acetylcholine receptor (AChR). A major antigen for these antibodies is the Ca2+ release channel of the sarcoplasmic reticulum the ryanodine receptor (RyR). These antibodies are found mainly in MG patients with a thymoma MG and correlate with severe MG symptoms. The antibodies recognize a region near the N-terminus on the RyR, which seems to be of importance for RyR regulation. The antibodies cause allosteric inhibition of RyR function in vitro, inhibiting Ca2+ release from sarcoplasmic reticulum. PMID- 11412152 TI - Mitochondrial myopathies. AB - The first description of a patient with mitochondrial myopathy and deficient respiratory chain function was reported by Luft and coworkers almost 40 years ago. Subsequent studies in the 1970s and 1980s relied on a combination of morphological and biochemical methods to identify patients with mitochondrial disorders. However, the aetiology and pathogenesis remained largely unsolved and there was poor correlation between laboratory findings and clinical phenotypes. The fact that both mitochondrial DNA (mtDNA) and nuclear genes are necessary for the biogenesis of the respiratory chain, suggested that mutations of either genome might cause mitochondrial myopathy. This prediction has been verified during the last decade and pathogenic mutations of both genomes have been identified. This rapid accumulation of genetic information has lead to many good correlations between genotype and phenotype in mitochondrial disorders. The challenge for the future will be to elucidate molecular details of pathogenic processes and to develop effective treatments for patients with respiratory chain dysfunction. PMID- 11412153 TI - Isolated systolic hypertension: pressure on the anaesthetist? PMID- 11412154 TI - The effects of rigid collar placement on intracranial and cerebral perfusion pressures. AB - Rigid collars are routinely used to immobilise the cervical spine during early management of trauma victims until spinal injury is excluded. Spinal injuries commonly coexist in patients with severe head injury, and there is still uncertainty as to whether application of a rigid collar may adversely affect intracranial pressure. The aim of this study was to examine this effect by applying rigid collars to patients with traumatic head injury. The results showed a significant rise from the baseline intracranial pressure when the collars were applied (mean rise = 4.6 mmHg, p < 0.0001). The mean rise in intracranial pressure was greater in those patients with a baseline intracranial pressure > 15 mmHg compared to those with a baseline intracranial pressure < 15 mmHg (p < 0.05). Since there was no significant change in cardiorespiratory parameters during this time, venous compression in the neck seems a likely explanation for the effect observed. These findings suggest that in head-injured patients, rigid collars should be removed as soon as cervical spine injury has been excluded or, if this is delayed, an alternative method of spinal stabilisation considered. PMID- 11412155 TI - Effects of shivering prevention on haemodynamic and metabolic demands in hypothermic postoperative neurosurgical patients. AB - We evaluated the haemodynamic and metabolic effects of prevention of shivering after prophylactic nefopam administration in neurosurgical patients undergoing craniotomy and mild systemic hypothermia (33-35 degrees C). Forty patients were enrolled in a randomised, double-blind study. Before extubation, patients received intravenously either nefopam 0.12 mg.kg-1 or an equal volume of saline 0.9%. Left ventricular systolic work index, oxygen consumption index and systemic lactate concentration were recorded before, immediately after and every 20 min for 2 h after extubation. Shivering appeared in two patients treated with nefopam and in all control patients (p < 0.001). Both left ventricular systolic work index and oxygen consumption index were similar in the two groups before extubation, increased after extubation, and further increased in control patients showing a statistical difference compared to patients treated with nefopam. Our results suggest that nefopam is effective in preventing postoperative shivering in patients undergoing neurosurgery and mild hypothermia and attenuates the haemodynamic effects of shivering during rewarming. PMID- 11412156 TI - Analgesic efficacy and safety of nefopam vs. propacetamol following hepatic resection. AB - In order to compare the morphine-sparing effect, analgesic efficacy and tolerance of nefopam and propacetamol given at their highest recommended doses, 120 patients undergoing elective hepatic resection were randomly assigned to receive postoperative intravenous patient-controlled analgesia with morphine alone, or in combination with nefopam (20 mg.4 h-1) or propacetamol (2 g.6 h-1). Compared with the control group (43 [7-92] mg), median [range] cumulative morphine consumption for 24 h after the study started was halved in the nefopam group (21 [3-78] mg, p <0.001) and 20% lower in the propacetamol group (35 [6-84] mg, p = 0.15). Analgesia was superior in the nefopam group despite the lower morphine consumption. Adverse effects were comparable in the three groups, except for significantly more nausea in the control group (39% vs. 17 and 26% in the nefopam and propacetamol groups, respectively) and more sweating in the nefopam group (17% vs. 0 and 3% in the control and propacetamol groups, respectively). Overall patient satisfaction was better (p < 0.001) in patients given nefopam (97%) than those receiving morphine alone (82%) or propacetamol (74%). PMID- 11412157 TI - Minimum local analgesic dose of plain ropivacaine vs. ropivacaine combined with sufentanil during epidural analgesia for labour. AB - We have used the up-and-down allocation technique to assess the relative analgesic potencies of epidural ropivacaine alone and ropivacaine combined with sufentanil 0.75 microg.ml-1 in 42 women requesting epidural analgesia in the first stage of labour. Parturients were randomly allocated to one of the two epidural solutions in a double-blind manner. The concentration of local anaesthetic was determined by the response of the previous parturient: an effective concentration (pain < or = 10 mm on a 10-cm visual analogue pain score within 30 min) resulted in a 0.01% decrease in the concentration of ropivacaine for the next parturient, an ineffective concentration resulted in a 0.01% increase. Minimum local analgesic concentration of ropivacaine alone was 0.13% (95% CI 0.12-0.13%) compared with 0.09% (95% CI 0.08-0.1%) for ropivacaine with sufentanil (p < 0.00001). PMID- 11412158 TI - Quantitative physical chemistry analysis of acid-base disorders in critically ill patients. AB - Compared with the Henderson-Hasselbalch approach, the Stewart approach may better describe the mechanisms of acid-base physiology and disorders. We prospectively examined the acid-base disorders of 100 routine blood samples from critically ill patients using Stewart's physical chemistry analysis. The median results were pH 7.45, PaCO2 5.5 kPa, bicarbonate 27.2 mmol.l-1 and base excess 3 mmol.l-1. The median reference strong ion difference was 46.0 meq.l-1 and the measured median was 45.5 meq.l-1. The median reference total weak-acid concentration was 11.1 mmol.l-1. The measured median total weak-acid concentration was 6.8 mmol.l-1. From Stewart's approach, the most likely explanation for the overall alkalosis was decreased total weak-acid concentration resulting from decreased plasma albumin concentration. PMID- 11412159 TI - The arterial tourniquet: pathophysiological consequences and anaesthetic implications. AB - The arterial tourniquet is widely used in upper and lower extremity surgery and in intravenous regional anaesthesia. The local and systemic physiological effects and the anaesthetic implications are reviewed. Localised complications result from either tissue compression beneath the cuff or tissue ischaemia distal to the tourniquet. Systemic effects are related to the inflation or deflation of the tourniquet. Safe working guidelines for the application of an arterial tourniquet have not been clearly defined. PMID- 11412160 TI - The in vitro performance of carbon dioxide absorbents with and without strong alkali apparatus. AB - We report the in vitro longevity of a conventional soda lime carbon dioxide absorbent and an absorbent free from strong alkali (Amsorb). Although the times taken to breakthrough of carbon dioxide (> 0.5%) within an in vitro low flow breathing system were shorter with the alkali-free absorbent, we found that the size and shape of the absorbent container was the major factor in determining the efficiency of the carbon dioxide absorbents. PMID- 11412161 TI - Extracting arterial flow waveforms from pulse oximeter waveforms apparatus. AB - A method is described which allows an approximation to the arterial flow waveform to be derived from a pulse oximeter waveform. The observed pulse oximeter waveform is the sum of arterial inflow and venous outflow. These components are separated mathematically. Subtraction of the venous outflow reveals the underlying arterial flow waveform. The assumptions on which the method is based are stated explicitly and discussed. PMID- 11412162 TI - Arterial flow waveforms from pulse oximetry compared with measured Doppler flow waveforms apparatus. AB - This study compared derived arterial flow waveforms, extracted from pulse oximeter waveforms, with Doppler flow waveforms. Fingertip pulse oximeter waveforms and radial artery Doppler flow waveforms were measured simultaneously in volunteers. The pulse oximeter waveforms were processed to extract the arterial flow waveforms and these were compared with the measured Doppler waveforms. They were very similar. PMID- 11412163 TI - Neuromuscular monitoring in myasthenic syndrome. AB - We describe the anaesthetic management of a 72-year-old man with myasthenic syndrome. Pre-operatively, he was treated with 3,4-diaminopyridine and showed a strong hand grip. During general anaesthesia with nitrous oxide and sevoflurane in oxygen, a mechanomyograph and two accelerographs were set up for the hands and left foot to monitor neuromuscular function. Insufficient force and acceleration of contraction with 1 Hz stimulation was observed in the hands. In the foot, the twitches produced by 1 Hz and train-of-four stimulation could barely be detected using the accelerograph, and the train-of-four ratio fluctuated between 70 and 100%. No neuromuscular blocking drugs were used during surgery. After discontinuation of sevoflurane, responses to train-of-four stimulation remained small, but a strong response to tetanic stimulation was observed, with post tetanic facilitation. Extubation was successful, and recovery from anaesthesia was uneventful. Tetanic stimulation and post-tetanic facilitation are important in monitoring neuromuscular function in patients with myasthenic syndrome whose train-of-four responses are insufficient. PMID- 11412164 TI - Epidural anaesthesia for Caesarean section in a patient with Devic's Syndrome. AB - The anaesthetic management of a 29-year-old paraplegic woman suffering from Devic's Syndrome scheduled to undergo Caesarean section under epidural anaesthesia is presented. The case is discussed with particular reference to the risk of autonomic hyperreflexia. PMID- 11412165 TI - Arterial blood sampling practices in intensive care units in England and Wales. AB - A postal survey of arterial blood sampling practices in 280 intensive care units throughout England and Wales found that very few measures are taken to reduce diagnostic blood loss in adult intensive care patients from arterial sampling. The average volume of blood withdrawn to clear the arterial line before sampling is 3.2 ml; subsequently returned to the patient in only 18.4% of intensive care units. Specific measures to reduce the blood sample size by the routine use of paediatric sample tubes in adult patients occurs in only 9.3% of intensive care units. In paediatric units, the average volume withdrawn was 1.9 ml and this was routinely returned in 67% of units. Some aspects of arterial blood sampling practices identified in this survey may contribute to iatrogenic anaemia in intensive care patients. PMID- 11412166 TI - Pathological evaluation of venous emboli during total hip arthroplasty. AB - One of the possible causes of bone cement implantation syndrome during total hip arthroplasty is pulmonary embolism, as has been suggested by a characteristic image seen during transoesophageal echocardiography, the so-called 'Snow Flurry'. However, the nature of the embolic material has not been definitively determined. We studied seven patients undergoing cemented or uncemented total hip arthroplasty. 'Snow Flurry' images were detected in all patients. Blood was aspirated from the right atrium during 'Snow Flurries' and was subjected to pathological examination. Amorphous, eosinophilic, fine granular material was seen in all specimens. The material did not originate from cement. Fat and bone marrow were not detected. The material seen may consist of fine particles of bone ('bone dust') originating from reamed bone. PMID- 11412167 TI - The effects of remifentanil on haemodynamic stability during rigid bronchoscopy. AB - We examined the effect of remifentanil on the haemodynamic response to rigid bronchoscopy in 22 adult ASA 2-4 patients, randomly allocated to receive 1.0 microg.kg-1 remifentanil over 1 min followed by 0.5 microg.kg-1.min-1 (remifentanil group) or 2.0 microg.kg-1 fentanyl followed by a saline infusion (control group). Following the initial infusion, all patients received a sleep dose of propofol followed by rocuronium 0.6 mg.kg-1 and their lungs were ventilated using a Sander's injector attached to the rigid bronchoscope. Direct arterial blood pressures, heart rate and ST segment changes on the CM5 lead configuration of the electrocardiograph were measured at 1-min intervals from before induction of anaesthesia until the end of bronchoscopy. Arterial pressures and heart rate were similar in the two groups before and after induction of anaesthesia. Remifentanil attenuated the haemodynamic response to bronchoscopy (p < 0.05 for increases in arterial pressures and heart rate). Four patients in the remifentanil group had ST segment depression compared with eight patients in the control group, but this was not statistically significant. PMID- 11412168 TI - Royal College of Anaesthetists Tutors. A survey of their duties and resources. AB - Royal College of Anaesthetists Tutors have a key role in the delivery of postgraduate anaesthetic training in the UK. We report the results of a postal questionnaire sent to all College Tutors in April 2000. An 89% response rate (253 of 283 Tutors) was received. Respondents were responsible for a median 11 trainees. Forty-nine Tutors with responsibility for larger numbers of trainees had the assistance of a deputy. The duties of a College Tutor comprised the organisation and delivery of training, appraisal and assessment, and 73% of respondents undertook all of these. In addition, a proportion had other administrative duties. The average time spent on College Tutor duties was 1.6 notional half days. Almost two-thirds of Tutors received some resources (time or remuneration) for their duties. PMID- 11412169 TI - New variant Creutzfeldt-Jakob disease--is our practice safe? PMID- 11412170 TI - The case report is not dead. PMID- 11412171 TI - The case report--an endangered species. PMID- 11412172 TI - Luer-lock epidural connections. PMID- 11412173 TI - ASA classification of patient for organ donation. PMID- 11412174 TI - Undergraduate teaching; a missed opportunity? PMID- 11412175 TI - The anaesthetic logbook. PMID- 11412177 TI - Secondary transfer of intensive care patients by helicopter. PMID- 11412178 TI - Anaesthesia without induction rooms. PMID- 11412179 TI - Pre-operative visits by anaesthetists. PMID- 11412180 TI - Continuous patient monitoring. PMID- 11412182 TI - Another case of electromagnetic interference in the operating theatre. PMID- 11412183 TI - Relief of tension subcutaneous emphysema with a small-bore subcutaneous drain. PMID- 11412184 TI - Capnography can aid in diagnosis of tracheobronchial injury. PMID- 11412185 TI - Prone pulmonary artery catheter insertion aided by ultrasound. PMID- 11412186 TI - Midazolam pre-induction can save money. PMID- 11412187 TI - Serial failure of Diprifuser infusion pumps. PMID- 11412189 TI - More hidden leaks. PMID- 11412191 TI - Any which way but loose? --the significance of gas flow direction through a vaporiser. PMID- 11412193 TI - 'Puffer-PCA device' for rheumatoid arthritis patients. PMID- 11412195 TI - Bad connection--a missing dimension. PMID- 11412196 TI - Lumbar spondylodiscitis after epidural anaesthesia at a distant site. PMID- 11412197 TI - Informed consent, and fainting fathers. PMID- 11412198 TI - Dislodgement of epidural catheters in labour. PMID- 11412200 TI - Failed epidural in an obese patient--blame it on Pythagoras! PMID- 11412201 TI - Advantages and disadvantages of the Arrow FlexTip Plus epidural catheter. PMID- 11412202 TI - A new interest in an old remedy for headache and backache for our obstetric patients: a sphenopalatine ganglion block. PMID- 11412203 TI - High-dose intrathecal diamorphine for analgesia after Caesarean section. PMID- 11412205 TI - An audit of the pain management work done by anaesthetists during on-call time in a teaching hospital. PMID- 11412206 TI - Anaesthesia in an unusual location. PMID- 11412207 TI - Yet more ampoule spikes. PMID- 11412209 TI - An assessment of the immunological status of patients with renal cell carcinoma based on the relative abundance of T-helper 1- and -2 cytokine-producing CD4+ cells in peripheral blood. AB - OBJECTIVE: To assess the immunological status of patients with renal cell carcinoma (RCC), by analysing the proportion of cluster-of-differentiation 4 positive (CD4+) cells showing intracellular cytokine production, i.e. interferon gamma derived from T-helper (Th) 1 and interleukin-4 derived from Th2 cells, among peripheral blood lymphocytes from these patients Patients, subjects and methods Peripheral blood samples (5 mL) were collected from 36 patients (mean age 61 years, range 44-78) with RCC before and after they underwent nephrectomy. The proportion of cytokine-producing CD4+ cells was determined by flow cytometric analysis after stimulating the cells with phorbol 12-myristate 13-acetate, ionomycin and brefeldin A, and staining the cells with fluorescein isothiocyanate labelled anti-interferon-gamma, anti-interleukin-4 and anti-immunoglubulin-2b antibodies. The results were expressed as the percentage of cytokine-producing cells in the CD4+ population. As a control, peripheral blood obtained from 35 healthy volunteers (mean age 34 years, range 22-49) was also analysed. RESULTS: The proportion of CD4+ cells producing interferon-gamma and interleukin-4 was significantly higher (P < 0.04 and P < 0.001, respectively) in patients with RCC than in controls. The Th1/Th2 ratio (i.e. the ratio of CD4+ cells producing each cytokine) was significantly lower in patients with RCC (P < 0.001). There was a significant correlation in the controls between interferon-gamma and interleukin 4 production (r = 0.489, P < 0.01) but not in patients with RCC. The proportion of CD4+ cells producing interleukin-4 was significantly higher and the Th1/Th2 ratio significantly lower in patients with high-stage than in those with low stage RCC (P < 0.05). The percentage of CD4+ cells producing interleukin-4 was significantly less after nephrectomy in those with low-stage RCC (P < 0.01) and the Th1/Th2 ratio significantly greater (P < 0.05) than before nephrectomy; there was no such trend in patients with high-stage RCC. Conclusion An evaluation of the production of interferon-gamma and interleukin-4 in CD4+ peripheral blood lymphocytes is useful for assessing the immunological status of patients with RCC; there is a change in the predominant response from Th1 to Th2 with increasing stage of RCC. PMID- 11412210 TI - How widespread are the symptoms of an overactive bladder and how are they managed? A population-based prevalence study. AB - OBJECTIVE: To determine the prevalence of chronic and debilitating symptoms of the overactive bladder, defined here as the presence of chronic frequency, urgency and urge incontinence (either alone or in any combination), and presumed to be caused by involuntary detrusor contractions. Subjects and methods Data were collected using a population-based survey (conducted by telephone or direct interview) of men and women aged >/= 40 years, selected from the general population in France, Germany, Italy, Spain, Sweden and the United Kingdom, using a random stratified approach. The main outcome measures were: prevalence of urinary frequency (> 8 micturitions/24 h), urgency and urge incontinence; the proportion of participants who had sought medical advice for symptoms of an overactive bladder; and current or previous therapy received for these symptoms. RESULTS: In all, 16 776 interviews were conducted in the six European countries. The overall prevalence of overactive bladder symptoms in individuals aged >/= 40 years was 16.6%. Frequency (85%) was the most commonly reported symptom, followed by urgency (54%) and urge incontinence (36%). The prevalence of overactive bladder symptoms increased with advancing age. Overall, 60% of respondents with symptoms had consulted a doctor but only 27% were currently receiving treatment. Conclusion Symptoms of an overactive bladder, of which frequency and urgency are as bothersome as urge incontinence, are highly prevalent in the general population. However, only a few affected individuals currently receive treatment. Taken together, such findings indicate that there is considerable scope for improvement in terms of how physicians diagnose and treat this condition. PMID- 11412211 TI - Intravesical oxybutynin for treating incontinence resulting from an overactive detrusor. PMID- 11412212 TI - Capsaicin receptor VR1 and ATP-gated ion channel P2X3 in human urinary bladder. AB - OBJECTIVES: To determine the presence, distribution and molecular forms of the vanilloid receptor VR1, and confirm the presence and distribution of the ATP gated ion channel P2X3 in the human urinary bladder. Materials and methods Normal urinary bladder tissues were obtained at postmortem from four subjects. Eight urinary bladder biopsies were also taken from patients with detrusor hyper reflexia treated with intravesical resiniferatoxin. The specimens were studied using affinity-purified specific antibodies to VR1 and P2X3 by Western blotting and immunocytochemistry, and compared with immunostaining using antibodies to the pan-neuronal marker PGP 9.5 and Schwann cell marker S-100. RESULTS: VR1- and P2X3 immunoreactive fine nerve fibres were scattered throughout the suburothelium of the normal bladder and cystoscopic biopsies, and traversed the muscle layer. They had a similar distribution to PGP 9.5-immunoreactive fibres, but there were fewer, suggesting localization in subsets of axons. Western blot studies showed an expected 100-kDa VR1 protein and a P2X3-immunoreactive 66-kDa protein. Conclusion VR1 and P2X3 are present in the human urinary bladder and may contribute to distinct pathophysiological states of bladder overactivity, in accord with their differential expression in sensory neurones. Intravesical vanilloids act via VR1 and are effective in the treatment of detrusor hyper reflexia. P2X3 may represent a selective therapeutic target for other causes of overactive bladder. PMID- 11412213 TI - False-positive pregnancy tests in patients with enterocystoplasties. AB - OBJECTIVE: To determine the incidence and causes of false-positive pregnancy tests in patients with enterocystoplasties. Patients and methods In a pilot study, 15 consecutive patients (both sexes, with any type of urinary diversion) provided a urine sample for testing using the Clearview pregnancy test (Unipath Ltd, Bedford, England). False-positive results occurred only in patients with continent reservoirs; patients with ileal conduits produced no false-positive results and were excluded from the main study. The main study included 23 patients with a urinary reservoir constructed partly or solely of large bowel or ileum, who provided urine and blood samples for the assay of beta human chorionic gonadotrophin and protein. Each urine sample was also analysed with the Clearview test. RESULTS: In the pilot study, three men and two women had positive pregnancy test results. In the main study, 14 patients had a positive Clearview test (nine women and five men). One women was found to be pregnant and was therefore excluded from further analysis, giving a false-positive rate of 57%. There was no difference in the type of reservoir, urine biochemistry and urine analysis results between the positive and negative groups. Frozen/thawed samples all tested negatively. Conclusion It is possible that the mucus produced in enterocystoplasties may be interfering with the pregnancy test. Patients should be aware of this and not rely on commercial test kits. There is some potential that this test system could be adapted for measuring mucus production in urinary diversions. PMID- 11412214 TI - The serous-lined tunnel principle for urinary reconstruction: a more rational method. AB - OBJECTIVE: To examine the feasibility of using the serous-lined-tunnel principle for orthotopic neobladder, continent cutaneous diversion and ureteric replacement by an intestinal segment. Patients and methods We created: (i) an orthotopic ileal neobladder using the serous-lined technique for antirefluxing ureteric implantation in 16 patients; (ii) a continent ileal pouch, adopting the principle for continent-valve construction and for ureteric implantation, in 10 patients (another patient with a failed continent valve underwent revision using an adaptation of this principle; and (iii) by applying the same principle an ileal ureter with a proximal antirefluxing mechanism was constructed in two patients (with lower ureteric cancer), and total replacement of the ureter by a tubular segment of the colon in association with a continent transverse colon pouch in one irradiated patient. RESULTS: In all, 52 ureters implanted into ileal neobladders or continent pouches functioned well, with neither obstruction nor reflux; 11 continent valves functioned well with no incontinence. Two patients with ileal ureters showed no ileo-ureteric reflux and had less hydronephrosis than before surgery. The tubularized ureter provided a unidirectional flow into the pouch. Conclusion Ureteric reimplantation and continent valve formation achieved by adopting the serous-lined tunnel principle provided satisfactory results. The versatility of the principle is apparent in the present experience and the creative application of the serous-lined tunnel principle should be possible in urinary reconstruction. PMID- 11412215 TI - Finasteride and tamsulosin used in benign prostatic hypertrophy: a review of the prescription-event monitoring data. AB - OBJECTIVE: To review the results of non-interventional observational cohort studies of 14 772 patients treated with finasteride and 12 484 patients treated with tamsulosin, both studies being of national proportions and undertaken in general medical practice in England. METHODS: Both studies were undertaken by prescription-event monitoring (PEM), whereby the exposure data are derived from information provided in strict confidence by the Prescription Pricing Authority of the National Health Service. The outcome data are derived from 'green form' questionnaires completed by the prescribing general practitioners (GPs). Additional data are obtained by medical follow-up with the attending practitioners. Adverse experience was measured in three ways; as reports of events which the doctors considered to represent adverse drug reactions; as reports of reasons for stopping the drug; and by studying the incidence density of each reported event. For these purposes a computerized dictionary containing 1430 higher level terms was used. The duration of exposure in the finasteride study was approximately 1 year and was approximately 6 months in the tamsulosin study. RESULTS: The outcome data on the 14 772 and 12 484 patients treated in the finasteride and tamsulosin studies were derived from the 63% and 57.4% of the green forms sent out and returned, respectively. The finasteride cohort included two women and the tamsulosin cohort 70 women. The mean (SD) age of the men in the two cohorts was, respectively, 69.0 (9.2) and 66.2 (11.7) years. Both drugs were well tolerated on long-term therapy and 69.6% (10 274 patients) of the total finasteride and 62.0% (7739 patients) of the total tamsulosin cohort were still receiving the drug at the end of 6 months. In the finasteride study, impotence or ejaculatory failure was reported in 2.0% of the patients still receiving the drug; there were reports of decreased libido in 1.0% and gynaecomastia was reported whilst the drug was still being prescribed in 39 patients (0.3% of the cohort). With tamsulosin, uncommon cases of dizziness, headache, malaise and hypotension (89 reports in 12 484 patients, i.e. 0.7% of the cohort) were common to the findings of reported adverse reactions, reasons for stopping the drug and events of highest incidence density. None of the deaths which occurred in either of these large cohorts was attributed by either the reporting GPs or the PEM medical staff to the drugs examined. Conclusion The GPs rated the drugs effective in most patients; tolerance and adverse experience was consistent with the known pharmacology of the two drugs. No serious, unexpected adverse effects were identified. PMID- 11412216 TI - Consensus development of a histopathological classification system for chronic prostatic inflammation. AB - OBJECTIVE: To develop a standardized histopathological classification system for chronic prostatitis (standardized description of prostatic inflammatory infiltrates) based on a literature review, extensive prospective evaluations in two recognized prostatitis research centres and widespread consensus of international urological centres identified as having major expertise or interest in chronic prostatitis. METHODS: Relevant articles for review were identified by a Medline search undertaken by the Cochrane Review Group in Prostate Diseases and Urologic Malignancies, and cross-checking bibliographies of retrieved studies, reviews, book chapters and abstracts of the American Urological Association and International Prostatitis Collaborative Network Annual Meetings. Initial drafts were based on classification systems independently developed by the Prostatitis Research Centers at Queen's University in Canada and University of Washington in the USA. A collaborative draft was distributed to 20 urological/pathological clinical centres who participated in the North American Chronic Prostatitis Collaborative Research Network and First International Prostatitis Collaborative Network. A consensus classification system was then distributed to the participating panel for acceptance. RESULTS: The literature review identified a reasonably consistent description of inflammatory infiltrate locations and patterns that were further incorporated into the draft based on the Queen's University and University of Washington proposals. Eighteen (90%) of the identified Prostatitis Centers participated in the revision of the draft and the final consensus process. The final consensus document classifies prostatic inflammation according to its extent and grade/severity in each tissue compartment (location). Conclusion The consensus of the expert panel was that this classification system can be used in the evaluation of prostatic inflammation in prostate biopsies, transurethral resected prostate chips or prostatectomy specimens. A standardized accepted framework to describe histopathological prostate inflammation will prove useful in evaluating prostate disease. PMID- 11412217 TI - Combined androgen blockade in prostate cancer: meta-analyses and associated issues. PMID- 11412218 TI - Are dietary influences on the risk of prostate cancer mediated through the insulin-like growth factor system? AB - OBJECTIVES: To investigate whether dietary factors that appear to affect the risk of prostate cancer may be similarly associated with serum levels of insulin-like growth factor 1 (IGF-1). Patients and methods In the context of a case-control study, 112 men were admitted to three teaching hospitals in Athens, Greece, for disorders other than cancer. Sociodemographic data and detailed histories of smoking, alcohol and coffee consumption were recorded. A validated food-frequency questionnaire was administered by an interviewer and serological measurements of IGF-1 and its binding protein-3 conducted. RESULTS: IGF-1 declined significantly by almost 25% among men aged >75 years and there was a small reduction in IGF-1 levels with increased alcohol intake, with a mean (95% confidence interval, CI) change of -1.6 (- 2.2 to -0.9)% for an increment of one drink per day. There was no evidence for an effect of either smoking or coffee consumption on IGF-1 level. Among foods, the consumption of cooked tomatoes was substantially and significantly inversely associated with IGF-1 levels, with a mean (95% CI) change of -31.5 (- 49.1 to -7.9)% for an increment of one serving per day. CONCLUSIONS: The strongest known dietary risk factor for prostate cancer (lycopene deficit, as reflected in a reduced intake of cooked tomatoes) and an important endocrine factor in the aetiology of this disease (IGF-1) seem to be related in a way that suggests that at least one, and perhaps more, exogenous factors in the development of prostate cancer may be mediated through the IGF-1 system. PMID- 11412219 TI - The independent prognostic value of comorbidity among men aged < 75 years with localized prostate cancer: a population-based study. AB - OBJECTIVE: To investigate which prognostic factors apply in patients with localized prostate cancer diagnosed after the introduction of prostate-specific antigen (PSA) testing, as comorbidity has significant prognostic value for patients who were diagnosed with localized prostate cancer in the 1970s. Patients and methods Using the Eindhoven Cancer Registry, we assessed a population-based cohort of patients aged < 75 years with localized (T1-T3M0) prostate cancer diagnosed between 1993 and 1995 in a defined area with 2 million inhabitants in the southern Netherlands (n = 894). After a mean follow-up of 2.9 years, overall survival was modelled by Cox regression analyses. RESULTS: Comorbidity was the most important prognostic factor, especially for those aged < 70 years; at 60 years old, patients with one concomitant disease were twice as likely to die than those with no comorbidity (95% confidence interval, CI, 1.0-4.3), whereas the hazard ratio (HR) was 7.2 (3.1-16.6) for two or more diseases. This was not caused by a reduced use of curative treatment for these patients. At the age of 74 years, comorbidity was no longer a significant prognostic factor. Poor differentiation of the tumour was also an important prognostic factor at all ages; this became increasingly apparent 2 years after diagnosis (HR 3.4, CI 1.5 7.7). Conclusion Comorbidity had a decisive influence on the prognosis for patients with localized prostate cancer. Because this effect was stronger in younger patients the assessment of comorbidity seems most important when evaluating the risk of early death. PMID- 11412220 TI - Increased patient satisfaction from transrectal ultrasonography and biopsy under sedation. AB - OBJECTIVE: To determine the acceptability and patient satisfaction of transrectal biopsy undertaken with the patient under sedation. Patients and methods A retrospective questionnaire was sent to 100 patients who had undergone transrectal biopsy between January and August 1998. Levels of patient acceptability and satisfaction were assessed using visual analogue scales (VAS, with a maximum score of 10 being the least satisfactory or acceptable) and direct questions about the side-effects of the procedure. A subsequent prospective study was undertaken on 130 patients undergoing transrectal biopsy with sedation between January 1999 and January 2000. RESULTS: The mean score for patient discomfort with sedation was 1.5, compared with 3.5 with no sedation. The overall satisfaction score improved from 3.1 to 0.9 with sedation. Complication rates were comparable, although slightly higher overall in the prospective group. Conclusion Sedation can significantly reduce patient discomfort and make the transrectal biopsy a more satisfactory experience for the patient. This is particularly important in the proportion of men who need to be considered for repeat biopsies. PMID- 11412221 TI - The use of the diode laser for treating urethral strictures. AB - OBJECTIVE: To evaluate the efficacy of the diode laser as a new tool for treating urethral strictures. Patients and methods The study comprised 22 patients with urethral strictures; a diode laser was used to circumferentially ablate the scar tissue of the strictures. Through a cystoscope, a 400-600 micrometer semirigid fibre was used to transmit the laser energy in the direct-contact mode. The procedure was carried out with the patient under general or spinal anaesthesia, but the last three patients successfully tolerated local anaesthesia. A catheter was then left in place for 1-5 days. RESULTS: The patients were followed for a mean (range) of 26.7 (9-39) months; in the 14 with previously untreated strictures the procedure was successful in 11, with no recurrence during the follow-up. However, seven of the eight patients with a recurrent stricture after previous internal cold-knife urethrotomy had a further recurrence, but occurring at longer intervals than after the internal urethrotomy. Conclusion This experience with the diode laser for treating urethral strictures suggests that it is safe and reliable, especially as the first line of treatment. PMID- 11412222 TI - Histopathological mapping of open testicular biopsies in patients with unobstructive azoospermia. AB - OBJECTIVE: To evaluate, in patients with unobstructive azoospermia, the heterogeneity of spermatogenesis within the testes and thus whether there is any region of advanced spermatogenesis. Patients and methods Seventy infertile men (mean age 34 years, SD 5.01) with no varicoceles or testicular atrophy had bilateral open testicular biopsies taken from six different sites. For each biopsy specimen the number of seminiferous tubules and of tubules with sperm maturation were counted (by light microscopy at x 400). The ratio of tubules with active spermatogenesis to the total number was calculated for each biopsy sample. RESULTS: The mean (SD) right and left testicular volumes were 19.82 (7.8) and 18.84 (7.89) mL, respectively; the patients' follicle-stimulating hormone level was 8.34 (1.17) IU/mL. On sextant biopsy spermatozoa were detected in 42 of the 70 patients (60%). The mean (SD) ratio of tubules with spermatozoa was 5.23 (0.8)% for the right and 5.37 (0.76)% for the left testes. There was no statistically significant difference in the ratio of seminiferous tubules positive for spermatozoa at the different biopsy sites in either the right or left testis. Spermatozoa were identified in only one to three biopsy sites in almost half of those with maturation arrest; this ratio increased to 74% in patients diagnosed as having Sertoli-cell-only syndrome with focal spermatogenesis. Conclusion There is no region of the testis that is rich or advanced in spermatogenesis in patients with unobstructive azoospermia. Without multiple testicular biopsy it is possible to miss advanced spermatogenesis in some unobstructed patients. The sextant testis biopsy is a reliable method for detecting the presence and exact location of seminiferous tubules with spermatozoa in patients with unobstructive azoospermia. PMID- 11412223 TI - Vascular risk factors and erectile dysfunction. PMID- 11412224 TI - Returning a referral for erectile dysfunction to the referrer by two different routes. AB - OBJECTIVE: To compare the prescribing pattern and attitude of general practitioners (GPs) in response to a clinic returning a patient referred for erectile dysfunction (ED) to the referrer by two different methods. METHODS: Referrals on a waiting list for an assessment of ED were reviewed and a subgroup of patients with criteria enabling them to be eligible for a prescription under the National Health Service (NHS) identified. The GP was informed either in writing or by telephone that the clinic had written to the patient, suggesting he make direct contact with his GP. A follow-up questionnaire was sent to each GP one month after the initial letter to the patient and contact with the GP. RESULTS: Of 91 questionnaires sent to GPs, 66 (73%) were completed; an additional five GPs corresponded by letter rather than completing the questionnaire. The long waiting time for assessment had led to 35% of patients having already tried sildenafil, and by the time the questionnaire was completed, 57% of patients had tried sildenafil. Ten times as many referrers indicated that they were happy to initiate a prescription for sildenafil than not to do so, for those men eligible for an NHS prescription. More GPs who had received a letter returned the completed questionnaire (80%) than those who had received a courtesy telephone call (64%). There were no differences between the groups of GPs in their attitude to our contact with their patient and no difference in prescribing pattern. Conclusion The provision of guidelines and advice to GPs either by telephone or letter is acceptable practice in reducing long waiting-list times for ED. Safe, simple and effective treatments are available for GPs to use under NHS guidelines. PMID- 11412225 TI - Peyronie's disease-the Plymouth experience of extracorporeal shockwave treatment. AB - OBJECTIVE: To describe our experience of the use of extracorporeal shockwave treatment (EST) for Peyronie's disease. Patients and methods The study included 28 patients (mean age 57 years, range 34-72) with stable Peyronie's disease who were treated with 3.9 (3-5) sessions of EST to the Peyronie's plaque. The patients' erectile function, pain and penile angle were assessed before and after treatment with EST. RESULTS: Of the 28 patients, 20 felt that their erection improved after the procedure; 11 patients were able to recommence sexual intercourse and the index of erectile function increased in all but one patient. Conclusion EST produces a significant improvement in pain and penile angle, with no serious complications. PMID- 11412226 TI - The use of renal scintigraphy in assessing the potential for recovery in the obstructed renal tract in children. AB - OBJECTIVE: To assess the value of renal scintigraphy with 99mTc dimercaptosuccinic acid (DMSA) in predicting functional recovery after the surgical relief of obstructed kidneys in children. Patients and methods Forty three children underwent surgery to relieve upper urinary tract obstruction; 37 had pelvi-ureteric junction obstruction and six had vesico-ureteric junction obstruction. The indication for surgery was a combination of an obstructed renogram with symptoms of either pain or pyelonephritis. Most children (41) had < 40% function on the affected side before surgery, with just two having hyperfunction (> 55%). In all patients intravenous urography before surgery showed hydronephrosis, and a micturating cystogram was used to exclude coexisting reflux in the presence of an associated megaureter. Diuretic renography (using 99mTc-mercaptoacetyl triglycine or 123I-hippuran) and DMSA scintigraphy were both carried out before surgery and the renography repeated 6 months afterward. RESULTS: The renographic drainage curves improved from obstructed to unobstructed or 'dilated unobstructed' on all postoperative studies. Regression analysis showed that preoperative DMSA scan was an excellent predictor of outcome (P < 0.001) whilst the preoperative renogram was a relatively poor predictor of the functional result. In four patients where the initial renographic function was < 10%, DMSA scintigraphy predicted correctly the capacity for recovery in three and the inability to improve in the fourth. Conclusion Before surgery, DMSA scintigraphy in children with upper urinary tract obstruction is a more useful estimate of probable long-term renal function than value from diuresis renography. If there is doubt about the desirability of reconstructive surgery, a DMSA scan may eliminate the need for more invasive methods of estimating recovery, e.g. a period of nephrostomy drainage. PMID- 11412227 TI - Diversion colitis in children with colovaginoplasty. AB - OBJECTIVE: To highlight the occurrence of diversion colitis and its effects in colovaginoplasty. Patients and methods The records of 18 children who had undergone colovaginoplasty were reviewed retrospectively. Nine patients had androgen-insensitivity syndrome and three each had congenital adrenal hyperplasia, vaginal agenesis and Mayer-Rokitansky syndrome. Through an abdominoperineal approach a segment of sigmoid colon was isolated on its vascular pedicle and brought to the perineum in the plane between the urethra and rectum, as a blind pouch or as an interposition between the proximal vagina and the perineum. The mean (range) follow-up was 5 (1.5-8) years. RESULTS: There were no major complications during the early follow-up. Three patients developed severe vaginal discharge with bleeding 2-7 years after colovaginoplasty; examination showed erythema, oedema, ulceration and bleeding. Histology confirmed the classic features of diversion colitis in all three patients. One child responded to vaginal irrigation with a solution of short-chain fatty acids, and the other two were treated with steroid enemas and mesalazine after a poor response to short chain fatty acids. One of these patients has required surgical reduction of an excessively long neovagina. Conclusion Symptomatic diversion colitis can occur after colovaginoplasty. The severity of the symptoms raises concerns about the use of colovaginoplasty in children and alternative techniques of vaginal replacement should be considered in the first instance. PMID- 11412228 TI - The Yang-Monti ileovesicostomy: a problematic channel? AB - OBJECTIVE: To compare the differences in the quality of Mitrofanoff channels created using appendix and re-tubularized small bowel (the Yang-Monti ileovesicostomy). Patients and methods The case-notes were reviewed retrospectively for all patients who underwent a Mitrofanoff procedure using either appendix or small bowel, over a 5-year period from June 1994 to July 1999. RESULTS: In all, 92 patients underwent 94 Mitrofanoff procedures; the appendix was used in 69 and small bowel in 25. The underlying diagnoses were exstrophy epispadias complex (38), neuropathic bladder (21), anorectal malformations and cloacal anomalies (15), posterior urethral valves (nine) and miscellaneous (nine). The mean (range) age at operation was 9.2 (1.1-18.3) years. The mean (range) follow-up for the appendix group was 37 (6.7-65) months and for the Monti group 25 (6-66) months. Catheterization problems occurred in 18 (27%) patients from the appendix group; two needed an adjustment of technique, six dilatation and 10 revision. Stomal stenosis occurred in 10 (15%) patients, bladder level stenosis in four (6%) and conduit necrosis in two. Catheterization problems were reported in 15 (60%) patients from the Monti group; five needed revision, three dilatation and seven are being managed conservatively. The incidences of stomal stenosis (four, 16%) and bladder level stenosis (two, 8%) were comparable with the appendix group. In addition, two patients had distal channel (sub-stomal) stenosis and two had mid-channel stenosis. The problem unique to the Yang-Monti channel was a pouch-like dilatation in seven patients (28%), all of whom presented with catheterization problems; five are being managed conservatively and two have needed pouch resection. Stomal prolapse occurred in five (7%) patients in the appendix group, but in none of the Monti group. CONCLUSIONS: The appendix is the conduit of choice for a Mitrofanoff procedure. Re-tubularized small bowel conduits have a considerably higher incidence of catheterization problems. Anatomical factors may contribute to the unique incidence of pouch formation. PMID- 11412229 TI - The use of microvessel density in assessing human urological tumours. PMID- 11412230 TI - The prevalence of loss of heterozygosity in chromosome 3, including FHIT, in bladder cancer, using the fluorescent multiplex polymerase chain reaction. AB - OBJECTIVE: To determine some of the genetic alterations involved in the pathogenesis and progression of transitional cell carcinoma of the bladder. Materials and methods In a population-based study, freshly frozen tissue was collected from all patients newly diagnosed with urinary bladder cancer in the Stockholm region during 1995-1996. The prevalence of loss of heterozygosity (LOH) was assessed at seven sites on chromosome 3, analysed in 151 patients, using a fluorescent multiplex polymerase chain reaction based on DNA from the tumour and peripheral blood. RESULTS: LOH was detected in 12.1% (at 3q25-26.2) to 22.1% (at 3p11-12) of the informative cases. Relatively frequent LOH was detected at 3p22 24.2 (21.6%), at 3p14.2 within FHIT (21.5%), and at 3p11-12 (22.1%). Of 151 tumours, 72 (47.7%) showed LOH at one or more loci on chromosome 3. LOH on chromosome 3 was weakly associated with tumour grade (P = 0.095), but not with tumour stage (P = 0.701). However, when the frequency of LOH was analysed individually at each site, the prevalence of LOH at 3p11-12 was closely correlated with higher tumour stage (P = 0.011). Replication errors were detected in only four of 151 (2.6%) tumours. Conclusion These findings suggest that the 3p11-12 locus may involve a putative candidate tumour-suppressor gene which might be associated with bladder tumour invasiveness. The FHIT gene locus showed a relatively high frequency of LOH even in Ta tumours. PMID- 11412231 TI - The interaction of nitric oxide and prostaglandins in the control of corporal smooth muscle tone: evidence for production of a cyclooxygenase-derived endothelium-contracting factor. AB - OBJECTIVE: To investigate the interaction of endothelium-derived nitric oxide (NO) and prostaglandins (PGs) in regulating corporal smooth muscle tone in vitro. Materials and methods Strips of rabbit corpus cavernosum were mounted in organ chambers for the measurement of isometric tension. Strips were submaximally contracted with noradrenaline and concentration-response curves (CRCs) to acetylcholine (ACh) were constructed before and after treatment with 5 micromol/L atropine, 20 micromol of the cyclooxygenase inhibitor indomethacin and 10 micromol of the PGH2/thromboxane A2 receptor antagonist SQ29548. The NO synthase (NOS) inhibitors L-NG-monomethyl arginine (L-NMMA) and L-NG-nitroarginine (L NOARG) were added to strips at tonic tension in the presence and absence of indomethacin, and after this CRCs to ACh were constructed. RESULTS: The addition of ACh to strips produced a concentration-dependent relaxation which was inhibited by atropine. Indomethacin, but not SQ29548, significantly increased relaxation to ACh. Relaxation to ACh was impaired by L-NMMA, but adding ACh to strips treated with L-NOARG resulted in contractile responses, whilst both effects were reversed by indomethacin. L-NMMA and L-NOARG led to increases in tonic tone which were unaffected by indomethacin. CONCLUSIONS: In rabbit corpus cavernosum there is a tonic release of NO which does not appear to be inhibited by a vasoconstrictor prostanoid. Endothelium-dependent relaxation to ACh results in the dual production of NO and a cyclooxygenase-derived endothelium contracting factor which acts in opposition to NO; this factor is unlikely to act on PGH2/TXA2 receptors. PMID- 11412232 TI - Detection of chromosomal alterations in bladder cancer by comparative genomic hybridization. AB - OBJECTIVES: To characterize which genomic alterations occur in bladder cancers of different grades and stages, and to evaluate the prognostic implication of chromosomal imbalances. Materials and methods Twelve transitional cell carcinoma cell lines were used to study chromosomal aberrations, using comparative genomic hybridization to examine metaphase cells. RESULTS: There was a mean of 6.0 aberrations per tumour with 4.9 gains and 1.08 deletions per tumour cell line. The mean number of aberrations was the same in grade 2 and 3 tumours. High-stage (>/= T2) carcinomas had markedly more genomic gains than had low-stage (T1) tumours, with 7.2 gains and 1.3 deletions per >/= T2 tumour, and 2.7 gains and 0.8 deletions per T1 tumour, although the difference was not statistically significant. The most frequent changes were gains of chromosome 1 (seven), 9 and 16 (six), and losses of chromosomes 14 (three), 21 and Y (two). The changes 1p+, 3p+, 9q+, 14p-, 16q+ and Yp- were significantly more frequent in low-grade tumours, with 1p+, 3p+ and Yq- the most frequent in stage T1 tumours, and 16p+, 9q+ and 13q- the most frequent in stage >/= T2 tumours. Conclusion These findings indicate several genomic regions in chromosomes 1, 9, 14 and 16 that may carry genes for the development and progression of bladder cancers. PMID- 11412233 TI - Transposing the urethral meatus in the treatment of recurrent and postcoital cystitis in women with hypospadias. PMID- 11412234 TI - A novel use of the testis after radical inguinal lymphadenectomy. PMID- 11412235 TI - Ruptured renal artery aneurysm mimicking hydronephrosis in pregnancy. PMID- 11412236 TI - Congenital seminal vesicle cysts: an unusual but treatable cause of lower urinary tract/genital symptoms. PMID- 11412237 TI - 5-Hydroxytryptamine-induced potentiation of cholinergic responses to electrical field stimulation in pig detrusor muscle. PMID- 11412238 TI - A prospective study of conservatively managed acute urinary retention: prostate size matters. PMID- 11412240 TI - Editor's comment on: "Press release: further research supports Viagra safety profile". PMID- 11412244 TI - Ethics in surgical practice. PMID- 11412245 TI - Resection margins in Crohn's disease. PMID- 11412246 TI - Haemobilia. AB - BACKGROUND: This review aims to establish whether increased use of invasive procedures and the trend toward conservative management of major trauma has resulted in an increased incidence of haemobilia. METHOD: A Medline (http://igm.nlm.nih.gov/)-based search of the English language literature from January 1996 to December 1999 inclusive was performed using the keywords haemobilia, hemobilia, haematobilia and hematobilia. The presentation, aetiology, investigation, management and outcome of 222 cases were reviewed. RESULTS: Two thirds of cases were iatrogenic while accidental trauma accounted for 5 per cent. Haemobilia may be major, constituting life-threatening haemorrhage, or minor; it may present many weeks after the initial injury. Diagnosis is most commonly confirmed by angiography. Management is aimed at stopping bleeding and relieving biliary obstruction; 43 per cent of cases were managed conservatively and 36 per cent were managed by transarterial embolization (TAE). Surgery was indicated when laparotomy was performed for other reasons and for failed TAE. The mortality rate was 5 per cent. CONCLUSIONS: Although the incidence of iatrogenic haemobilia has risen considerably, the bleeding is often minor and can be managed conservatively. When more urgent intervention is required, TAE is usually the treatment of choice. There is no evidence that the conservative management of accidental liver trauma increases the risk of haemobilia. PMID- 11412247 TI - Systematic review of antiplatelet therapy for the prevention of myocardial infarction, stroke or vascular death in patients with peripheral vascular disease. AB - BACKGROUND: Antiplatelet agents may prevent vascular events and death in patients with peripheral vascular disease (PVD). METHODS: A systematic review of 39 randomized controlled trials of antiplatelet therapy in patients with PVD was performed. RESULTS: For patients with PVD the number suffering a non-fatal myocardial infarction, non-fatal stroke or vascular death in the antiplatelet group was 6.5 per cent compared with 8.1 per cent in the placebo group (odds ratio 0.78 (95 per cent confidence interval (c.i.) 0.63--0.96); P = 0.02), favouring antiplatelet treatment. For infrainguinal bypass surgery (ten trials) and balloon angioplasty (two) the differences were still in favour of antiplatelet therapy, but they did not reach statistical significance. In five trials of aspirin against another antiplatelet agent, 8.4 per cent in the aspirin group suffered a vascular event compared with 6.6 per cent in the second antiplatelet group (odds ratio 0.76 (95 per cent c.i. 0.64--0.91); P < 0.01), favouring ticlopidine/clopidogrel/aspirin + dipyridamole against aspirin alone. CONCLUSION: Antiplatelet therapy reduces serious vascular events and vascular death in patients with PVD. For infrainguinal arterial surgery or balloon angioplasty the benefit remains unproven, but the number of trials to date is small. There is also evidence to support the use of antiplatelet drugs other than aspirin for the prevention of vascular events in those with PVD. PMID- 11412248 TI - Randomized clinical trial of the effect of open versus laparoscopically assisted colectomy on systemic immunity in patients with colorectal cancer. AB - BACKGROUND: Laparoscopic surgery is believed to produce an attenuated metabolic stress response and to have a less dampening effect on the immune response than open surgery. To date, the effect has not been studied in a randomized clinical trial of colorectal cancer. METHODS: The study was a two-armed randomized prospective trial conducted in parallel with the UK Medical Research Council's Conventional versus Laparoscopic-Assisted Surgery in Colorectal Cancer (CLASICC) trial comparing laparoscopically assisted colorectal surgery for left-sided tumours with conventional open surgery. Systemic immunity was assessed by determining the T- and B-cell counts, the CD4 : CD8 ratio, the natural killer cell counts, the immunoglobulin (Ig) G, IgM and IgA levels, and C3 and C4 levels. The white cell phagocytic activity (nitroblue tetrazolium test) was studied before operation and on the third postoperative day. RESULTS: A total of 236 patients were randomized in the immune study between 11 March 1997 and 14 August 1999; 161 had complete preoperative and postoperative assays for the analysis of results. There was no difference in mean response between the two surgical groups for each of the immune parameters studied. The unadjusted difference for the primary endpoint, T-cell count, 3 days after operation was - 1.6 per cent (95 per cent confidence interval - 5.0 to 1.8 per cent). CONCLUSION: There is no difference in the systemic immune response in patients having laparoscopically assisted colectomy compared with those undergoing conventional open surgery for colorectal cancer. PMID- 11412249 TI - Management of 155 patients with benign liver tumours. AB - BACKGROUND: Benign hepatic tumours continue to represent a diagnostic and therapeutic challenge. This study evaluates the indications and results of resection compared with observation in patients with benign hepatic tumours. METHODS: Patients with a primary diagnosis of benign liver tumour were identified from a prospective database and evaluated retrospectively. RESULTS: From January 1992 to June 1999, 155 patients with benign hepatic tumours were evaluated. The diagnoses included haemangioma (n = 97), focal nodular hyperplasia (FNH) (n = 42), hepatic adenoma (n = 12) and cystadenoma (n = 4). Sixty-eight patients (44 per cent) underwent resection because of symptoms (n = 36), inability to exclude a malignancy (n = 31) or enlargement on serial imaging (n = 11). The operative morbidity and mortality rates were 21 per cent and zero respectively. Thirty patients had a preoperative percutaneous needle biopsy, 19 of which were either incorrect or indeterminate. Overall, 39 of 42 patients with symptoms attributed to the tumour were asymptomatic after resection and 18 of 21 patients with symptoms considered unrelated to the tumour were asymptomatic after a period of observation and/or treatment of unrelated conditions (median follow-up 16 months). CONCLUSION: When indicated, resection of benign liver tumours can be performed safely. Symptomatic patients with a small FNH or haemangioma can be observed because their symptoms are unlikely to be related to the liver tumour. Percutaneous needle biopsy rarely changes management. PMID- 11412250 TI - Botulinum toxin injection is an effective treatment for axillary hyperhidrosis. PMID- 11412251 TI - Microcirculatory effects of experimental acute limb ischaemia-reperfusion. AB - BACKGROUND: The object of this study was to develop an animal model in which changes in microvascular haemodynamics and leucocyte-vessel wall interactions due to acute limb ischaemia-reperfusion (I/R) can be measured in the skin. Furthermore, it was investigated whether these changes are related to local muscle injury. METHODS: Male Lewis rats were subjected to unilateral limb ischaemia for 1 h (n = 8) or 2 h (n = 8) by cuff inflation, or to a sham protocol (n = 6). Intravital video microscopic measurements of leucocyte-vessel wall interactions, venular diameter, red blood cell velocity and reduced velocity (which is proportional to wall shear rate) were performed in skin venules before ischaemia and at 0.5, 1, 2, 3 and 4 h after the start of reperfusion. Oedema and leucocyte infiltration of ischaemic/reperfused skeletal muscle were quantified histologically. RESULTS: In skin venules, both 1 and 2 h of ischaemia induced a significant increase in leucocyte rolling (six and five times baseline, respectively; P < 0.05) and adherence during reperfusion (eight and four times baseline; P < 0.05). No significant increase in muscular leucocyte infiltration was detected. After an initial hyperaemic response of 180 per cent of baseline values (P < 0.05), blood flow decreased to about 60 per cent after 4 h of reperfusion in skin venules of both experimental groups. I/R induced tibial muscle oedema, the severity of which depended on the ischaemic interval (wet to dry ratio: control, 4.0; 1 h, 4.5 (P not significant); 2 h, 5.8 (P < 0.05)). CONCLUSION: A non-invasive animal model was developed that enables investigation of the consequences of acute limb I/R. PMID- 11412252 TI - Recombinant soluble P-selectin glycoprotein ligand 1 moderates local and remote injuries following experimental lower-torso ischaemia. AB - BACKGROUND: A central role for the polymorphonuclear leucocyte (PMN) in skeletal muscle ischaemia-reperfusion has been demonstrated by the observation that PMN depletion reduced local and remote pulmonary vascular permeability. This study investigated the role of recombinant soluble P-selectin glycoprotein ligand immunoglobulin fusion protein (rPSGL-Ig), a P- and E-selectin antagonist, in moderating injury. METHODS: Mice underwent 2 h of hindlimb ischaemia and 3 h of reperfusion. Muscle and lung vascular permeability index (PI) was assessed by extravasation of (125)I-radiolabelled albumin. Lung myelo peroxidase (MPO) activity was also measured. RESULTS: In mice treated with rPSGL-Ig 1 mg/kg before reperfusion (n = 12) muscle PI was reduced by 40 per cent, whereas it was moderated by 20 per cent in animals treated 30 min after reperfusion (n = 15). Lung PI in mice treated with rPSGL-Ig before (n = 12) and 30 min after (n = 15) reperfusion was reduced by over 99 and 98 per cent respectively. Lung MPO activity in mice treated with rPSGL-Ig before (n = 10) and 30 min after (n = 12) reperfusion was reduced by 68 and 58 per cent respectively. Treatment with rPSGL Ig 1 h after reperfusion, or with m20ek.Fc 1 mg/kg (n = 9; negative control for rPSGL-Ig which is inactive for selectin binding) before reperfusion failed significantly to moderate local or remote organ injury. CONCLUSION: Selectin blockade moderated local skeletal muscle and remote lung injury following hindlimb ischaemia--reperfusion. Significantly, delayed antiselectin therapy also decreased injury. PMID- 11412253 TI - Use of minilaparotomy in the treatment of colonic cancer. AB - BACKGROUND: The feasibility and safety of a minilaparotomy approach for curative resection of colonic cancer have not been fully elucidated. The purpose of this study was to compare outcomes utilizing this alternative technique with those of conventional laparotomy. METHODS: Eighty-four patients scheduled to undergo resection for colonic cancer via minilaparotomy between 1997 and 1999 were studied prospectively. The minilaparotomy involved complete resection performed through a skin incision less than 7 cm in length. Sixty-nine patients who underwent a similar resection via a conventional laparotomy between 1994 and 1996 served as the control group. RESULTS: The minilaparotomy approach was successful in 72 of 84 patients. Colectomy type, operating time and histopathological features of tumours were similar between cases and controls, whereas operative blood loss in the control group was significantly greater (P = 0.002). Postoperative times to standing, walking, passage of flatus and urinary catheter removal were significantly shorter in the minilaparotomy group (P = 0.007, P = 0.003, P = 0.03 and P = 0.006 respectively), and analgesic requirements were significantly lower (P = 0.001). At a median follow-up of 24.8 months there have been no tumour recurrences at the minilaparotomy incision sites. CONCLUSION: A minilaparotomy approach to the curative resection of colonic cancer is an attractive alternative to conventional laparotomy in selected patients. PMID- 11412254 TI - Randomized study to examine the value of on-table inspection of the appendix mucosa. PMID- 11412255 TI - Tumour location and the effects of preoperative radiotherapy in the treatment of rectal cancer. AB - BACKGROUND: Preoperative radiotherapy improves local control and survival in rectal cancer, but may also increase postoperative morbidity and mortality rates. Establishing selection criteria for preoperative radiotherapy is crucial. The tumour level above the anus may be one such criterion. The effect of preoperative radiotherapy in relation to the distance between the tumour and the anus was therefore assessed. METHODS: In 457 patients operated for cure included in the Stockholm II Trial the local recurrence rate in irradiated and non-irradiated patients was analysed in relation to the tumour location (low, mid or upper rectum). RESULTS: Radiotherapy reduced the local recurrence rate from 30 to 20 per cent in low rectal cancer, from 25 to 11 per cent in mid rectal cancer and from 21 to 5 per cent for tumours in the upper rectum. CONCLUSION: With conventional surgical techniques preoperative radiotherapy plays an important role in rectal cancer irrespective of the location of the tumour. To irradiate only patients with tumours in the lower rectum and to omit this treatment for patients with tumours in the mid and upper rectum cannot be recommended. Whether this statement is valid with standardized total mesorectal excision (TME) surgery is not known. Until this knowledge is available the current indications for preoperative radiotherapy should probably also be used with TME surgery. PMID- 11412256 TI - Long-term results of treating hepatic colorectal metastases with cryosurgery. AB - BACKGROUND: The purpose of this study was to determine the long-term efficacy of cryosurgery as an adjunct to hepatic resection in patients with colorectal liver metastases not amenable to resection alone. METHODS: Thirty patients met the following inclusion criteria: metastases confined to the liver and judged irresectable, ten or fewer metastases, cryosurgery alone or in combination with hepatic resection allowed tumour clearance. RESULTS: Median follow-up was 26 (range 9--73) months. Overall 1- and 2-year survival rates were 76 and 61 per cent respectively. Median survival was 32 months. Disease-free survival at 1 year was 35 per cent, at 2 years 7 per cent. Six patients developed recurrence at the site of cryosurgery; given that the total number of cryosurgery-treated lesions was 69 the local recurrence rate was 9 per cent. CONCLUSION: In patients with colorectal liver metastases, local ablative techniques can be used as an effective adjunct to hepatic resection to obtain tumour clearance. PMID- 11412257 TI - Follow-up confirms sustained benefit of circumferential stapled anoplasty in the management of prolapsing haemorrhoids. AB - BACKGROUND: Circumferential stapled anoplasty is gaining popularity as a safe and effective treatment in the surgical management of haemorrhoids and mucosal prolapse. However, little is known about the medium- and long-term durability of this procedure. The aim of this study was to evaluate the medium-term results of stapled anoplasty in maintaining symptom remission and to identify possible procedure-related adverse effects. METHODS: Of a consecutive series of 85 stapled anoplasties in 83 patients, 64 patients were eligible for 6-month review; 50 patients were seen in surgical outpatient clinics and seven were contacted by telephone. All patients attending the outpatient clinic were questioned about current symptoms and overall satisfaction with the procedure. The staple line was palpated digitally and inspected at proctoscopy. RESULTS: Median (interquartile range) symptom scores were 6 (5--8) before operation compared with 0 (0--1) at 6 months (P < 0.01). There was no deterioration in symptoms between 6-week and 6 month follow-up. No recurrences and no procedure-related adverse effects, in particular impaired continence or persistent anal pain, were identified. CONCLUSION: The initial promising results of circumferential stapled anoplasty in effectively treating haemorrhoidal symptoms appear to be sustained at 6-month follow-up. PMID- 11412258 TI - Endocoil magnetic resonance imaging quantification of external anal sphincter atrophy. AB - BACKGROUND: Anal function depends on the integrity and quality of the sphincter muscles. The diagnosis of external anal sphincter atrophy on endocoil magnetic resonography has been associated with poor outcome from sphincter repair, although the imaging criteria for atrophy remain unclear. METHODS: Women with intact sphincters on endosonography and either normal (more than 60 cm H(2)O) (n = 9) or low (n = 16) squeeze pressures had endocoil magnetic resonography and electromyography. The area and fat content of the external anal sphincter and puborectalis were measured on mid-coronal magnetic resonography and images were graded as showing normal, intermediate or advanced atrophy. The definition of the external anal sphincter on endosonography and the thickness of the internal anal sphincter were also assessed. RESULTS: Women with a normal anal squeeze pressure had a larger external anal sphincter cross-sectional area (mean(s.d.) 240(56) versus 193(62) mm(2); P = 0.01) with a lower mean fat content (mean(s.d.) 23(4) versus 30(6) per cent; P < 0.001) than those with low squeeze pressures. There was an overall correlation between squeeze pressure, cross-sectional area (r = 0.32, P = 0.02) and fat content (r = - 0.51, P < 0.001). Patients with a thin (less than 2 mm) internal anal sphincter and/or a poorly defined external sphincter on endosonography were more likely to have atrophy (positive predictive value 74 per cent). CONCLUSION: : Potential endosonographic markers for external anal sphincter atrophy are suggested, and a visual scale for endocoil magnetic resonographic assessment has been validated. PMID- 11412259 TI - Probability of axillary node involvement in patients with tubular carcinoma of the breast. AB - BACKGROUND: The aim of this study was to investigate the frequency of axillary metastasis in women with tubular carcinoma (TC) of the breast. METHODS: Women who underwent axillary dissection for TC in the Western Sydney area (1984--1995) were identified retrospectively through a search of computerized records. A centralized pathology review was performed and tumours were classified as pure tubular (22) or mixed tubular (nine), on the basis of the invasive component containing 90 per cent or more, or 75--90 per cent tubule formation respectively. A Medline search of the literature was undertaken to compile a collective series (20 studies with a total of 680 patients) to address the frequency of nodal involvement in TC. A quantitative meta-analysis was used to combine the results of these studies. RESULTS: The overall frequency of nodal metastasis was five of 31 (16 per cent); one of 22 pure tubular and four of nine mixed tumours (P = 0.019). None of the tumours with a diameter of 10 mm or less (n = 16) had nodal metastasis compared with five of 15 larger tumours (P = 0.018). The meta-analysis of 680 women showed an overall frequency of nodal metastasis in TC of 13.8 (95 per cent confidence interval 9.3-18.3) per cent. The frequency of nodal involvement was 6.6 (1.7--11.4) per cent in pure TC (n = 244) and 25.0 (12.5- 37.6) per cent in mixed TC (n = 149). CONCLUSION: A case may be made for observing the clinically negative axilla in women with a small TC (10 mm or less in diameter). PMID- 11412260 TI - Prediction of prognosis by echocardiography in patients with midgut carcinoid syndrome. AB - BACKGROUND: The association between malignant midgut carcinoid tumours and right sided cardiac lesions is well known, but the pathogenetic link between tumour secretion and valvular disease is still obscure. The purpose of this investigation was to describe the morphological and functional changes of valvular heart disease in a large patient series and to correlate these findings with hormonal secretion and prognosis. METHODS: Of 64 consecutive patients with the midgut carcinoid syndrome followed between 1985 and 1998, valvular heart disease was evaluated in 52 patients by two-dimensional echocardiography, Doppler estimation of valvular regurgitation and flow profiles. A majority was also evaluated with exercise electrocardiography and spirometry. RESULTS: Structural and functional abnormalities of the tricuspid valve were found in 65 per cent of patients, while only 19 per cent had pulmonary valve regurgitation. Long-term survival was related to excessive urinary excretion of 5-hydroxyindole acetic acid of over 500 micromol in 24 h, but the main predictor of prognosis was the presence of severe structural and functional abnormalities of the tricuspid valve. Although advanced tricuspid abnormalities were prevalent in this series, only one patient died from right ventricular heart failure. CONCLUSION: Tricuspid valvular disease is a common manifestation of the midgut carcinoid syndrome and advanced changes are associated with poor long-term survival. Active surgical and medical therapy of the tumour disease reduced the hormonal secretion and, combined with cardiological surveillance, made right ventricular heart failure a rare cause of death in these patients. PMID- 11412261 TI - Risk factors for complications following resection of large gastric cancer. AB - BACKGROUND: Although there is a low mortality rate after gastrectomy in Japan, most studies include many early gastric cancers. There have been few studies on the morbidity after gastrectomy for advanced gastric cancer. The aim of this study was to clarify the characteristics and risk factors for postoperative complications after resection of large gastric cancers based on three clinical factors: patient, operation and tumour. METHODS: A retrospective study was carried out on 97 patients with a gastric tumour measuring 10 cm or more in diameter. Postoperative complications were recorded and the patients were divided into two groups: 38 with complications and 59 without. Patient, operative and tumour findings were compared between the two groups. RESULTS: Overall morbidity and mortality rates were 39 and 7 per cent respectively. The most frequent complication was pleural effusion (17 per cent), followed by anastomotic leakage (14 per cent), abdominal abscess (12 per cent), wound infection (12 per cent), pancreatic leakage (8 per cent) and peritonitis (6 per cent). Risk factors associated with postoperative complications were operating time (400 versus 337 min, P < 0.01), blood loss (1338 versus 782 ml, P < 0.01), pancreatic invasion (26 versus 8 per cent, P < 0.05) and raised serum carcinoembryonic antigen (CEA) level (5 ng/ml or greater) (36 versus 17 per cent, P < 0.05), independent of patient age, nutritional status, type of gastrectomy, splenectomy or pancreatectomy, extent of lymph node dissection, tumour location, size and stage of disease. CONCLUSION: Even in Japan, the morbidity of gastrectomy for large gastric cancer is high and associated with operating time, blood loss, pancreatic invasion and serum CEA level. PMID- 11412262 TI - Endotoxin, cytokines and lipid peroxides in children with intussusception. AB - BACKGROUND: Intussusception is a relatively common paediatric surgical emergency. The aim of this study was to investigate selected inflammatory mediators in children with acute intussusception and to identify potentially useful plasma markers of clinical outcome. METHODS: Clinical, radiographic, operative and pathological details were recorded prospectively of all children presenting to a single institution with a confirmed diagnosis of acute intussusception during 1 year. Paired acute and convalescent venous blood samples were collected in a standard manner for blinded analysis of the following: malondialdehyde, C reactive protein (CRP), interleukin (IL) 6, neopterin, tumour necrosis factor alpha, endotoxin, and immunoglobulin (Ig) G and IgM antiendotoxin core antibody (EndoCAb). RESULTS: Thirty-two consecutive children (23 boys, nine girls) with a median age of 4 months were studied. Acute ileocolic intussusception was managed by air enema reduction (n = 19), operative reduction (n = 8) or surgical resection (n = 5). Peripheral blood cultures were sterile. Acute levels of plasma IL-6, neopterin and CRP were significantly raised in comparison to both normal laboratory ranges and convalescent samples (P < 0.001). Using stepwise discriminant analysis, CRP was identified as the best variable at distinguishing between the three treatment groups (P < 0.001). IgM EndoCAb concentrations were significantly greater in the convalescent sera of all the patients (P < 0.001). CONCLUSION: Acute ileocolic intussusception in childhood is associated with endotoxinaemia and significantly raised levels of circulating inflammatory cytokines. Plasma CRP at diagnosis showed a statistically significant positive correlation with disease severity. PMID- 11412263 TI - Totally extraperitoneal endoscopic repair of recurrent inguinal hernia. AB - BACKGROUND: Conventional repair of recurrent inguinal hernia is associated with a re-recurrence rate as high as 35 per cent. Endoscopic mesh repair has promising results regarding both recurrence and complication rates. METHODS: In a retrospective review, the results of endoscopic totally extraperitoneal repair were evaluated in 104 patients with 108 recurrent hernias. Follow-up was at least 1 year. Type of recurrence, time of occurrence after previous repair, duration of surgery, complications, duration of hospital stay and number of re-recurrences were evaluated. RESULTS: Follow-up ranged from 12 to 29 (mean 16) months. Forty three recurrences were direct, 41 indirect and 15 combined; one was a femoral hernia. Median time to previous operation was 36 months (range 8 days to 42 years). Median duration of surgery was 63 (range 25--160) min. While there were no complications during operation, 12 patients (12 per cent) had a postoperative complication. Two direct re-recurrences (2 per cent) occurred as a result of inadequate positioning of the prosthetic mesh. CONCLUSION: The endoscopic totally extraperitoneal technique is safe and effective for the repair of recurrent inguinal hernia. PMID- 11412264 TI - Major intra-abdominal pathology missed at laparoscopic cholecystectomy (Br J Surg 2000; 87: 1434-5). PMID- 11412265 TI - Chronic dysphagia following laparoscopic fundoplication (Br J Surg 2000; 87: 1341 5). PMID- 11412266 TI - Short-term results of femoropopliteal subintimal angioplasty (Br J Surg 2000; 87: 1361-5). PMID- 11412267 TI - Anatomy of Denonvilliers' fascia and pelvic nerves, impotence, and implications for the colorectal surgeon (Br J Surg 2000; 87: 1288-99). PMID- 11412268 TI - Vascular access for haemodialysis (Br J Surg 2000; 87: 1300-15). PMID- 11412269 TI - Vascular access for haemodialysis (Br J Surg 2000; 87: 1300-15). PMID- 11412270 TI - Role of resection and primary anastomosis of the left colon in the presence of peritonitis (Br J Surg 2000; 87: 1580-4). PMID- 11412271 TI - Suitability for endovascular aneurysm repair in an unselected population (Br J Surg 2001; 88: 77-81). PMID- 11412272 TI - Probiotics (Br J Surg 2001; 88: 161-2). PMID- 11412274 TI - Prospective evaluation of the morbidity of axillary clearance for breast cancer (Br J Surg 2001; 88: 114-7). PMID- 11412275 TI - Jejunal interposition to prevent postgastrectomy syndromes (Br J Surg 2000; 87: 1576-9). PMID- 11412277 TI - Adipocyte metabolism and the metabolic syndrome. PMID- 11412278 TI - Therapeutic considerations for postprandial dyslipidaemia. PMID- 11412279 TI - Clustering of cardiovascular risk factors with diabetes in Chinese patients: the effects of sex and hyperinsulinaemia. AB - OBJECTIVE: This study was designed to investigate factors which affect the clustering of cardiovascular risk factors with diabetes in Chinese patients. RESEARCH DESIGN AND METHODS: Six hundred and fifty-four patients with diabetes were assessed comprehensively for diabetes complications and cardiovascular risk factors in a metropolitan hospital in Beijing, China. Insulin resistance and secretion were also evaluated by measurement of glucose and insulin levels before and after a meal tolerance test. Results were analysed according to patient groups stratified by the number of cardiovascular risk factors coexisting with diabetes. RESULTS: Cardiovascular risk factors were common in Chinese diabetic patients. The clustering of three or more of these factors with diabetes occurred more often than by chance alone and was associated with postprandial hyperinsulinaemia. Patients with a high number of risk factors were more prone to macrovascular events but did not have higher albuminuria. Using the commonly adopted lower threshold for diagnosing obesity and central obesity in women, there were more women with multiple risk factors. However, this disappeared if the same criteria were used for men and women. Even in the presence of diabetes, cardiovascular risk factors were inadequately controlled in most patients. CONCLUSIONS: The concurrence of diabetes and other cardiovascular risk factors which constitute the metabolic syndrome is a common phenomenon in urban Chinese diabetic patients. It is associated with hyperinsulinaemia and possibly the female sex. This study emphasises the importance of public health measures to control cardiovascular risk factors in patients with diabetes. PMID- 11412280 TI - Comparison of adipose tissue changes following administration of rosiglitazone in the dog and rat. AB - Rosiglitazone (BRL-49653-C), a thiazolidinedione, is a potent agonist for the nuclear hormone receptor peroxisome proliferator-activated receptor gamma (PPARgamma). Thiazolidinediones have been reported to induce adipocyte differentiation in vitro and there is limited data on their effects in vivo. The objective of this study was to compare the effects of rosiglitazone on adipocyte differentiation between dogs and rats. Morphological (light and ultrastructural) and morphometric evaluations were conducted on perirenal adipose tissue from dogs that have been treated for 1 month with 0.4, 5, 60 mg/kg/day and rats treated for the same period with 80 mg/kg/day. There was a dose-related change in the phenotype of white adipose tissue in dogs, reflected by an increase in nuclear numerical density (up to threefold) and cytoplasmic area fraction (up to 2.1 fold). In addition, there was an enlargement of the nuclei and a reduction in the size of the white adipocyte lipid vacuoles. Ultrastructural changes included an increase in the number of mitochondria per adipocyte. In the rat, similar changes were seen in nuclear numerical density (1.5-fold increase) and cytoplasmic area fraction (2.2-fold increase). There were also increased numbers of mitochondria per cell in white adipocytes giving them similar numbers of mitochondria to brown adipocytes. In the brown adipocytes, there was a reduction in cytoplasmic area fraction with a corresponding increase in the size of the lipid filled vacuoles in other words there was a converging of the phenotypes of the white and brown adipose tissues. PMID- 11412281 TI - Insulin-sensitizing action of rosiglitazone is enhanced by preventing hyperphagia. AB - AIM: We investigated whether pair-feeding to prevent hyperphagia would potentiate the insulin-sensitizing effect of rosiglitazone in chow-fed and insulin-resistant dietary obese rats, and studied the role of leptin and hypothalamic neuropeptide Y as mediators of weight gain during treatment. METHODS: Dietary obese and chow fed rats (575 +/- 10 vs. 536 +/- 7 g; p < 0.01) were given rosiglitazone (30 mg/kg p.o.) or vehicle daily for 14 days. RESULTS: Energy intake and weight gain were greater in rosiglitazone-treated ad-lib-fed rats (body weight: chow + 24 +/- 2 g, rosiglitazone-treated + 55 +/- 2 g, p < 0.001; dietary obese + 34 +/- 2 g, rosiglitazone-treated + 74 +/- 7 g, p < 0.001). Half of each rosiglitazone treated group were pair-fed to vehicle-treated controls. Rosiglitazone normalized circulating free fatty acids (FFAs) and insulin sensitivity in dietary obese rats (homeostasis model assessment (HOMA): chow-fed controls, 3.9 +/- 0.3; dietary obese controls, 6.7 +/- 0.7; rosiglitazone-treated, ad lib-fed dietary obese, 4.2 +/- 0.5; both p < 0.01). Insulin sensitivity improved further with pair-feeding (HOMA: 2.9 +/- 0.4; p < 0.05 vs. rosiglitazone-treated, ad lib-fed dietary obese), despite unchanged FFAs. Qualitatively similar findings were made in chow fed rats. Pair-feeding prevented rosiglitazone-related weight gain in chow-fed, but not dietary obese rats (body weight: + 49 +/- 5 g, p < 0.001 vs. untreated dietary obese controls). Adipose tissue OB mRNA was elevated in dietary obese rats, reduced 49% (p < 0.01) by rosiglitazone treatment, and further (by 16%) with pair-feeding (p < 0.0001). Plasma leptin, however, only fell in the pair-fed group. Hypothalamic neuropeptide Y mRNA was unchanged throughout, suggesting that weight gain associated with high-dose rosiglitazone treatment is independent of hypothalamic neuropeptide Y. CONCLUSIONS: Food restriction potentiates the insulin-sensitizing effect of rosiglitazone in rats, and this effect is independent of a fall in FFAs. PMID- 11412282 TI - Influence of a combination of herbs on appetite suppression and weight loss in rats. AB - AIM: We evaluated the ability of a Chinese herbal formulation previously associated with weight loss to influence appetite and weight loss in a carefully controlled laboratory study performed on rats. As a secondary gain, results with this herbal formulation were compared with those from a commonly available phenylpropanolamine (PPA) compound. DESIGN: Eight rats were placed in each arm of a three-arm study, a total of 24 rats. All rats were gavaged with a 2-ml fluid volume containing no addition (control) or the two test substances (combined herbs or PPA) for the first 4 days of the week over 6 consecutive weeks; no gavages were given over weekends. Rats in the two-test groups were given a relatively low dose of the test substance for 3 weeks, followed by a higher dose over the next 3-week periods. Food and water intake were measured for 24-h periods over the ensuing week days. The average daily values for food and water intake for an individual rat were calculated on the basis of collected data over each 3-week period. The mean values for each rat obtained over the low- and high dose periods comprised results from averaging at least 10 measurements. RESULTS: Average daily food intake was decreased only with the herbal formulation, not the PPA compound at the low and high doses. Both the PPA compound and the herbal formulation lowered water intake significantly at the low and high doses. Rats ingesting the herbal formulation at the lower dose had statistically significant lower daily body weight changes over the 3 weeks than those ingesting the PPA compound. At the higher dose, body weight changes for both agents were significantly less than the control, but not significantly different from each other. No evidence of toxicity was seen in the blood chemistries or after histopathological examination. CONCLUSIONS: Data collected on rats suggest that the herbal formulation examined might be a useful and safe combination to overcome the overweight state and obesity. PMID- 11412283 TI - Hormone replacement therapy for postmenopausal women with diabetes. AB - BACKGROUND: Recent studies have shown only 17% of postmenopausal women in the UK receive prescriptions for hormone replacement therapy (HRT), with diabetic women being prescribed HRT 30% less often than non-diabetic women. Up to 80% of women experience menopausal symptoms, however, so it is expected that by using HRT many would gain benefit from symptom relief, in addition to benefiting from its effects in disease prophylaxis. METHODS: We conducted a questionnaire survey of the attitudes to HRT prescribing of 178 local general practitioners (GPs) and 47 hospital doctors, working in diabetes and gynaecology. RESULTS: Responses were received from 98 (55%) GPs and 39 (83%) hospital doctors. One hundred and thirty six (99%) would advise HRT for menopausal symptoms, with 133 (97%) advising HRT for a spontaneous premature menopause and 126 (92%) for a surgically induced menopause. One hundred and thirty-four (98%) would advise HRT for diabetic women with menopausal symptoms, with 122 (89%) indicating that HRT had no adverse effect on glycaemic control. For disease prophylaxis, all hospital doctors and 93 (95%) GPs would advise HRT for prevention of osteoporosis, although only 91 (93%) GPs would advise HRT to diabetic women for this indication. Thirty-three (85%) hospital doctors would advise HRT as prophylaxis against cardiovascular disease compared with 66 (67%) of GPs. For this indication in diabetic women, 32 (82%) hospital doctors and 67 (68%) GPs would advise HRT. Oral HRT was the most widely recommended preparation although GPs also frequently recommended patches. Topical gel, implants and other preparations were less commonly advised. Hospital doctors predominantly advised HRT for menopausal symptoms to be continued for between 1 and 5 years, although more GPs recommended HRT to be continued for longer than 5 years. The majority of doctors from both groups advised that HRT used for disease prophylaxis be continued for more than 5 years. CONCLUSION: More doctors than anticipated indicated they would recommend HRT, especially to diabetic women. Concerns among doctors about the side-effects of HRT and, in particular, the metabolic effects of HRT, do not appear to deter them from advising HRT. Willingness to prescribe a drug, however, is different from actually prescribing it and responses to questionnaires tend to be those regarded as being acceptable. Low patient demand for HRT due to fear or lack of awareness may also be contributing. Organizational change with the development of more menopause clinics may facilitate wider use of HRT. PMID- 11412284 TI - Lactic acidosis in metformin therapy: searching for a link with metformin in reports of 'metformin-associated lactic acidosis'. AB - OBJECTIVE: The link between metformin and lactic acidosis in metformin therapy may be causal, associated or coincidental. Our objective was to investigate this link by studying and analysing published reports of so-called 'metformin associated lactic acidosis'. RESEARCH DESIGN AND METHODS: systematically searched in the BIOSIS, DERWENT, EMBASE, MEDLINE, and PASCAL databases of the English language and non-English language literature for all reports of so-called 'metformin-associated lactic acidosis' published from May 1995 through January 2000. We did not include reports related to metformin overdose or contrast media induced renal failure. Metformin accumulation and concurrent pathologies were critically reviewed as precipitating factors for metformin-associated lactic acidosis. Metformin accumulation was assessed in terms of the recorded measurement of metformin concentration in plasma or, if not available, by the presence of primary renal failure, i.e. renal failure that was not secondary to a shock syndrome. RESULTS: We found 21 reports describing a total of 26 patients. Criteria of lactic acidosis (lactate > 5 mmol/l, pH or = 26 microg mL-1) as well as duration of exposure-dependent (> or = 30 min) increases in histidine, isoleucine, leucine, valine, methionine, serine, glycine, threonine, and decreases in glutamine, glutamate, aspartate, asparagine, arginine, ornithine, citrulline, alanine and taurine were observed (P < or = 0.05). Concerning PMN immune functions, methohexital significantly decreased O2-, H2O2 formation and MPO (> or = 26 microg mL-1, > or = 30 min, P < or = 0.05). CONCLUSIONS: Altogether, there is significant relevance to the pharmacological regimens which enhance the supply of methohexital in whole blood. In regards to our results, we suggest that considerable changes in PMN 'dynamic free amino acid pool', for example induced by methohexital, may be one of the determinants in cell nutrition adversely affecting PMN metabolism. It is partially through its effect on the PMN free amino acid pool that maleficent pharmacological stress may have an unintentional influence on PMN immune functions. PMID- 11412290 TI - Sevoflurane potentiates and blocks GABA-induced currents through recombinant alpha1beta2gamma2 GABAA receptors: implications for an enhanced GABAergic transmission. AB - BACKGROUND AND OBJECTIVE: The gamma-aminobutyric acidA receptor (GABAAR) is a target for anaesthetic agents. We investigated the interactions of sevoflurane with a recombinant GABAAR. Emphasis was on the mechanism of block, as relevant open-channel block by a volatile anaesthetic would possibly explain prolonged GABAergic postsynaptic currents. METHODS: The effect of sevoflurane on GABA induced currents through recombinant alpha1beta2gamma2 GABAAR channels was studied (patch clamp; HEK293 cells). GABA 0.01 mM or 1 mM was applied alone or together with sevoflurane (0.05 mM to 5 mM). RESULTS: Currents elicited by GABA 0.01 mM were increased by low sevoflurane concentrations to 183% and decreased by high sevoflurane concentrations (> 1 mM) to 34% (P < 0.05). Ten- to 90%-rise times of the currents were reduced by sevoflurane concentration dependently. At GABA (1 mM), peak currents and 10-90%-rise times decreased with increasing sevoflurane concentrations. A transient current increase was induced by discontinuation of GABA and sevoflurane. Such rebound currents indicate a reversal of an open-channel block by sevoflurane. CONCLUSIONS: Sevoflurane (a) increases the apparent affinity of GABA to the GABAAR, as suggested by the decreased current rise times. This explains the enhancement of the currents induced by low GABA concentrations (0.01 mM). Additionally, sevoflurane (b) induces a picrotoxin-like open-channel block at the GABAAR. The reversal of the open-channel block elicits a delayed GABA response. These findings indicate at least two different sites of action of sevoflurane at this receptor that are both important for an enhanced GABAergic synaptic transmission. PMID- 11412291 TI - The effect of different doses of propofol on tracheal intubating conditions without muscle relaxant in children. AB - BACKGROUND AND OBJECTIVE: Situations may occur in anaesthetic practice where the use of neuromuscular blocking drugs is unsuitable or contraindicated. We investigated the use of propofol given 5 min after fentanyl to permit endotracheal intubation in children. METHODS: We studied the intubating conditions and cardiovascular parameters in 60 ASA I and II children. Intravenous midazolam (0.1 mg kg(-1)) was given as premedication 5 min before the induction of anaesthesia. The children received different doses of propofol (group I, 2.5 mg kg(-1); group II, 3.0 mg kg(-1); group III, 3.5 mg kg-1) preceded by fentanyl (3.0 microg kg(-1)) given 5 min earlier. No neuromuscular blocking agents were administered. The intubating conditions were assessed using a four-point scoring system based on the degree of difficulty of laryngoscopy, the position of vocal cords and the intensity of coughing. RESULTS: Tracheal intubating conditions were adequate in 20% of the patients in group I, in 75% of the patients in group II and in 80% of the patients in group III (P < 0.05 for group I vs. groups II and III). Haemodynamic changes were not significantly different between the groups. CONCLUSIONS: Propofol (3.0 mg kg(-1)) preceded by fentanyl (3.0 microg kg(-1)) was adequate for the induction of anaesthesia in children and provided adequate tracheal intubating conditions without significant haemodynamic changes. This method represents a useful alternative technique for tracheal intubation when neuromuscular blocking drugs are contraindicated or should be avoided. PMID- 11412292 TI - A randomized controlled double-blind trial comparing piritramide and morphine for analgesia after hysterectomy. AB - BACKGROUND AND OBJECTIVE: Efficacy and side-effects of piritramide (pirinitramide) and morphine, given intravenously for postoperative analgesia after hysterectomy, were compared in a randomized controlled double-blind trial in 92 ASA class I-III patients. METHODS: Administration was investigator controlled during the first 90 min and subsequently via a patient-controlled device. Visual analogue scales for pain intensity and verbal rating scales for side-effects were taken repeatedly. RESULTS: Median visual analogue scores for pain intensity on a 100-mm scale 4, 8 and 24 h after surgery were 10, 8.5 and 5 mm in the piritramide group and 18, 10 and 8.5 mm in the morphine group. These differences are neither statistically nor clinically significant. Median values for nausea on a verbal rating scale from 0 to 3 were zero for both groups at all times with similar ranges. There was no difference in number of episodes of vomiting and retching and usage of antiemetics. The mean amount of piritramide used for initial titration was 15.2 mg; the respective amount of morphine was 15.4 mg. CONCLUSIONS: In this setting the two agents are equally effective and show a similar profile of side-effects. PMID- 11412293 TI - Acute respiratory acidosis does not increase plasma potassium in normokalaemic anaesthetized patients. A controlled randomized trial. AB - BACKGROUND AND OBJECTIVE: Few and conflicting data are available regarding the changes of plasma potassium concentration during acute respiratory acidosis in human beings. This study compares the acute changes in plasma potassium concentration in acutely hypercapnic patients and in non-hypercapnic patients during general anaesthesia. METHODS: Thirty-three patients undergoing interventional rigid bronchoscopy were studied. Ventilation of the lungs was randomly conducted using either spontaneous-assisted ventilation or intermittent negative-pressure ventilation. All patients received the same anaesthetic protocol. Arterial blood gases and osmolality, and plasma concentrations of glucose, sodium, potassium and chloride were measured. RESULTS: Intraoperatively, PaCO2 was higher during spontaneous-assisted ventilation than during intermittent negative-pressure ventilation (9 +/- 1.8 vs. 5.4 +/- 1.2 kPa, P < 0.001) and the pH was also lower during spontaneous-assisted ventilation than during intermittent negative-pressure ventilation (7.24 +/- 0.07 vs. 7.4 +/- 0.08, P < 0.001). Plasma potassium concentration remained similar in both groups (3.8 +/- 0.2 mmol L(-1) with spontaneous-assisted ventilation vs. 3.7 +/- 0.4 mmol L(-1) with intermittent negative-pressure ventilation). CONCLUSION: Acute respiratory acidosis does not affect plasma potassium concentration. PMID- 11412294 TI - Pressure-controlled inverse ratio ventilation after cardiac surgery. AB - BACKGROUND AND OBJECTIVE: Pressure-controlled inverse ratio ventilation was compared with controlled mechanical ventilation in patients after cardiac surgery. METHODS: Ten patients were ventilated after sternal closure using a Siemens Servo 900C ventilator to a target end-tidal PCO2 of 4.0 kPa. They were randomized to receive controlled mechanical ventilation or pressure-controlled inverse ratio ventilation. CO2-based data were recorded on a laptop personal computer, which together with arterial PCO2 permitted measurement of the respiratory dead space. Once measurements were complete the ventilator was switched to the other mode and new measurements taken. RESULTS: PaCO2 and VCO2 were virtually the same in both modes. Peak airway pressure (17.2 +/- 2.7 vs. 20.8 +/- 2.5 cmH2O, P < 0.01) and minute ventilation (4.9 +/- 1.1 vs. 5.3 +/- 1.1 cmH2O, P < 0.01) were less during pressure-controlled inverse ratio ventilation. Physiological dead space fraction (0.39 +/- 0.06 vs. 0.51 +/- 0.05, P < 0.001), airway dead space (56 +/- 15 vs. 81 +/- 15 mL, P < 0.001) and alveolar dead space fraction (0.25 +/- 0.07 vs. 0.31 +/- 0.09, P < 0.01) were all less during pressure-controlled inverse ratio ventilation. There were no differences in heart rate or mean arterial pressure. CONCLUSIONS: The prolonged inspiratory period and pressure-controlled flow pattern of pressure-controlled inverse ratio ventilation reduce the alveolar and airway dead spaces, and give lower peak airway pressures, compared with conventional ventilation, in cardiac surgical patients. PMID- 11412295 TI - Jet ventilation via a Univent tube for sleeve pneumonectomy. AB - A 51-year-old man presented with a right upper lobe adenocarcinoma with carinal extension. He underwent a right sleeve pneumonectomy, which involved a carinal resection with anastomosis between the trachea and left main bronchus. This report describes the successful use of jet ventilation, administered via the lumen of the bronchial blocker of a Univent tube. During 15 min of carinal resection, oxygenation of his left lung was maintained with the bronchial blocker bridging the airway discontinuity. PMID- 11412296 TI - Amniotic fluid embolism and isolated coagulopathy: atypical presentation of amniotic fluid embolism. AB - A 41-year-old multigravida presented at 32 weeks of gestation with polyhydramnios and an anencephalic fetus. Abnormal bleeding as a result of disseminated intravascular coagulation complicated an emergency Caesarean section for severe abdominal pain thought to be due to uterine rupture. Massive transfusion with blood products was necessary and the abdomen packed to control bleeding. The patient was transferred to the intensive care unit where she made a slow but complete recovery. Amniotic fluid embolism with atypical presentation of isolated coagulopathy is the likely diagnosis in this case. The case serves to demonstrate that amniotic fluid embolism may present with symptoms and signs other than the classical pattern of dyspnoea, cyanosis and hypotension. PMID- 11412297 TI - Anaesthesia for the professional singer. PMID- 11412298 TI - A difficult intubation made more difficult. PMID- 11412299 TI - Mechanisms of interleukin-10-mediated immune suppression. AB - Specific immune suppression and induction of anergy are essential processes in the regulation and circumvention of immune defence. Interleukin-10 (IL-10), a suppressor cytokine of T-cell proliferative and cytokine responses, plays a key regulatory role in tolerizing exogenous antigens during specific immunotherapy (SIT) of allergy and natural exposure to antigens. Specific T-cell tolerance is directed against the T-cell epitopes of an antigen and characterized by suppressed proliferative and T helper type 1 (Th1) and type 2 (Th2) cytokine responses. IL-10 elicits tolerance in T cells by selective inhibition of the CD28 co-stimulatory pathway and thereby controls suppression and development of antigen-specific immunity. IL-10 only inhibits T cells stimulated by low numbers of triggered T-cell receptors and which therefore depend on CD28 co-stimulation. T cells receiving a strong signal from the T-cell receptor alone, and thus not requiring CD28 co-stimulation, are not affected by IL-10. IL-10 inhibits CD28 tyrosine phosphorylation, the initial step of the CD28 signalling pathway, and consequently the phosphatidylinositol 3-kinase p85 binding to CD28. Together these results demonstrate that IL-10-induced selective inhibition of the CD28 co stimulatory pathway acts as a decisive mechanism in determining whether a T cell will contribute to an immune response or become anergic. PMID- 11412300 TI - Siglecs in the immune system. PMID- 11412301 TI - Studies on transcriptional regulation of the mucosal T-cell integrin alphaEbeta7 (CD103). AB - Integrin alphaEbeta7 is expressed almost exclusively by mucosal T cells and mucosal dendritic antigen-presenting cells (APCs) and is thought to be induced locally by transforming growth factor-beta (TGF-beta). In mice, mRNA for the alphaE subunit was found to be abundant in mucosal T cells but absent from other tissues. Exposure of a T-cell line to TGF-beta strongly up-regulated alphaE mRNA levels within 30 min, and nuclear run-on experiments established that regulation occurred at the level of transcription. The organization of the human alphaE gene and a very closely linked novel gene, ELG, was determined. The alphaE promoter was tested in T cells and fibroblasts and functioned equally well in both cell types and did not confer TGF-beta responsiveness. Regions of the promoter providing enhancer activity and phorbol 12-myristate 13-acetate (PMA) responsiveness were identified by deletion studies. DNAse 1 hypersensitivity analysis of 36 kb of the alphaE gene revealed one hypersensitive site, found only in alphaE+ cells, located near the transcription start points. These results show that, unlike the situation with other integrins, lineage specificity and cytokine responsiveness of alphaE transcription are not conferred by the proximal promoter. Specificity may depend on distant control elements that have not yet been identified. PMID- 11412302 TI - Preferential production of interferon-gamma by CD4+ T cells expressing the homing receptor integrin alpha4/beta7. AB - Recent studies indicate that T helper type 1 (Th1) and 2 (Th2) lymphocytes differ in their expression of molecules that control T-cell migration, including adhesion molecules and chemokine receptors. We investigated the relationship between cytokine production and expression of the homing receptor integrin alpha4/beta7 on T cells. We began by analysing cytokine production by human CD4+ CD45RA- memory/effector T cells following brief (4 hr) stimulation with phorbol 12-myristate 13-acetate (PMA) and ionomycin. alpha4/ beta7high CD4+ T cells were more likely to produce the Th1 cytokine interferon-gamma (IFN-gamma) than were alpha4/beta7- CD4+ T cells in all six subjects studied. In contrast, production of the Th2 cytokine interleukin-4 (IL-4) was similar on alpha4/ beta7high and alpha4/beta7- CD4+ T cells. In addition, we found that human CD4+ CD45RA- T cells that adhered to the alpha4/beta7 ligand mucosal addressin cell adhesion molecule 1 (MAdCAM-1) had a greater capacity to produce IFN-gamma than did non-adherent cells, suggesting that the association between alpha4/beta7 expression and IFN gamma production has functional significance. These results suggested that primary activation under Th1-promoting conditions might favour expression of alpha4/beta7. We directly examined this possibility, and found that naive murine CD4+ T cells activated under Th1-promoting conditions expressed higher levels of alpha4/beta7 compared to cells activated under Th2-promoting conditions. The association between alpha4/beta7 expression and IFN-gamma production by CD4+ T cells may help to determine the cytokine balance when MAdCAM-1 is expressed at sites of inflammation in the intestine or elsewhere. PMID- 11412303 TI - Specific immunoglobulin E responses in ZAP-70-deficient patients are mediated by Syk-dependent T-cell receptor signalling. AB - ZAP-70 deficiency is a rare primary immunodeficiency characterized by the absence of peripheral CD8+ T cells and defects in T-cell receptor (TCR) signalling. T cells in ZAP-70-deficient patients are assumed to have no helper functions for B cell immunoglobulin synthesis, whereas the patients rarely have antigen-specific antibodies. We experienced a ZAP-70-deficient patient, who had immunoglobulin E (IgE) antibodies specific to food allergens, and we investigated the mechanisms of switching to IgE in the patient. Peripheral blood mononuclear cells from the patient did not proliferate upon stimulation with the antigens but produced distinct levels of interleukin-4 (IL-4). Cell sorting analysis indicated that the cells that produced IL-4 in response to the antigens were enriched in CD4+ T cells. Purified CD4+ T cells from the patient produced IL-4 and expressed CD40L upon stimulation with anti-CD3. Moreover, CD4+ T cells pretreated with anti-CD3 induced mature epsilon transcript on naive B cells. Since the results indicated that there remained sufficient T-cell receptor (TCR)-signalling in the patient's T cells to exert antigen-specific IgE switching on B cells, we next investigated the expression of the ZAP-70-homologous kinase Syk. Syk was present in high levels in patient's CD4+ T cells and was tyrosine-phosphorylated after TCR stimulation. Inhibition of Syk by piceatannol resulted in decreased production of IL-4 and expression of CD40L on patient's CD4+ T cells. Moreover, Syk was expressed on all human T-cell leukaemia virus (HTLV-1)-transformed T-cell lines derived from peripheral blood of the patient, whereas it was low or undetectable in control lines. It was therefore concluded that specific IgE responses in the patient were most likely to be mediated by Syk-dependent TCR-signalling. PMID- 11412304 TI - Refocusing of B-cell responses following a single amino acid substitution in an antigen. AB - Intranasal immunization of BALB/c strain mice was carried out using baculovirus derived human chorionic gonadotrophin (hCG) beta-chain, together with Escherichia coli heat-labile enterotoxin. Gonadotrophin-reactive immunoglobulin A (IgA) was induced in a remote mucosal site, the lung, in addition to a systemic IgG response. The extensive sequence homology with luteinizing hormone (LH) results in the production of LH cross-reactive antibodies when holo-hCG is used as an immunogen. In contrast to wild-type hCGbeta, a mutated hCGbeta-chain containing an arginine to glutamic acid substitution at position 68 did not induce the production of antibodies which cross-react with LH. Furthermore, the epitopes utilized in the B-cell response to the mutated hCGbeta shifted away from the immunodominant region of the parent wild-type molecule towards epitopes within the normally weakly immunogenic C terminus. This shift in epitope usage was also seen following intramuscular immunization of rabbits. Thus, a single amino acid change, which does not disrupt the overall structure of the molecule, refocuses the immune response away from a disadvantageous cross-reactive epitope region and towards a normally weakly immunogenic but antigen-unique area. Similar mutational strategies for epitope-refocusing may be applicable to other vaccine candidate molecules. PMID- 11412305 TI - Third complementarity-determining region of mutated VH immunoglobulin genes contains shorter V, D, J, P, and N components than non-mutated genes. AB - The third complementarity-determining region (CDR3) of immunoglobulin variable genes for the heavy chain (VH) has been shown to be shorter in length in hypermutated antibodies than in non-hypermutated antibodies. To determine which components of CDR3 contribute to the shorter length, and if there is an effect of age on the length, we analysed 235 cDNA clones from human peripheral blood of VH6 genes rearranged to immunoglobulin M (IgM) constant genes. There was similar use of diversity (D) and joining (JH) gene segments between clones from young and old donors, and there was similar use of D segments among the mutated and non-mutated heavy chains. However, in the mutated heavy chains, there was increased use of shorter JH4 segments and decreased use of longer JH6 segments compared to the non mutated proteins. The overall length of CDR3 did not change with age within the mutated and non-mutated categories, but was significantly shorter by three amino acids in the mutated clones compared to the non-mutated clones. Analyses of the individual components that comprise CDR3 indicated that they were all shorter in the mutated clones. Thus, there were more nucleotides deleted from the ends of VH, D, and JH gene segments, and fewer P and N nucleotides added. The results suggest that B cells bearing immunoglobulin receptors with shorter CDR3s have been selected for binding to antigen. A smaller CDR3 may allow room in the antibody binding pocket for antigen to interact with CDRs 1 and 2 as well, so that as the VDJ gene undergoes hypermutation, substitutions in all three CDRs can further contribute to the binding energy. PMID- 11412306 TI - Short-term immunoglobulin A B-cell memory resides in intestinal lymphoid tissues but not in bone marrow of gnotobiotic pigs inoculated with Wa human rotavirus. AB - Immunological memory is important for protecting the host from reinfection. To investigate the development and sites of residence of intestinal memory B cells, and their role in protective immunity to reinfection with an enteric virus, we assessed the association between memory B cell and antibody-secreting cell (ASC) responses and protection using a gnotobiotic pig model for human rotavirus (HRV) infection and diarrhoea. The isotypes, quantities and tissue distribution of rotavirus-specific memory B cells and ASC were evaluated prechallenge (28 and 83 postinoculation days [PID]) and postchallenge (7 postchallenge days [PCD]), using enzyme-linked immunospot (ELISPOT) assay, in gnotobiotic pigs inoculated once with virulent or three times with attenuated HRV and challenged at PID 28 with the corresponding virulent HRV. Complete protection against HRV shedding and diarrhoea was associated with significantly higher numbers of immunoglobulin A (IgA) and immunoglobulin G (IgG) memory B cells and ASC in the ileum of virulent HRV-inoculated pigs at challenge. In contrast, pigs inoculated with attenuated HRV had lower numbers of IgA and IgG memory B cells and ASC in intestinal lymphoid tissues, but higher numbers in the spleen. The bone marrow had the lowest mean numbers of IgA and IgG memory B cells and ASC prechallenge in both groups of HRV-inoculated pigs. Therefore, bone marrow was not a site for IgA and IgG rotavirus-specific antibody production or for memory B cells after inoculation with live rotavirus, from 28 PID up to at least 83 PID. The effect of in vitro antigen dose was examined and it was determined to play an important role in the development of ASC from memory B cells for the different tissues examined. PMID- 11412307 TI - Immunoglobulin VH-gene usage of autoantibodies in mercuric chloride-induced membranous glomerulopathy in the rat. AB - Brown-Norway (BN) and Dorus Zadel Black (DZB) rats develop a T-cell-dependent membranous glomerulopathy (MGP) with high proteinuria and antiglomerular basement membrane (GBM) autoreactive antibodies (Abs), upon exposure to mercuric chloride (HgCl2). Laminin is an important autoantigenic target of the anti-GBM Abs, absorbing approximately 30% of the anti-GBM reactivity. Although many anti-GBM Abs have undergone isotype switching, it is currently unclear whether affinity maturation occurs during the HgCl2-induced autoimmune response. To address this question we analysed the rearranged immunoglobulin heavy chain variable-region genes (VHDJH regions) of 15 mAbs that were previously obtained from HgCl2-treated rats. Seven of these mAbs exhibit reactivity towards laminin. Our study showed that the VH-gene usage of antilaminin mAbs is largely restricted to the PC7183 VH gene family (six out of seven). In addition, we demonstrated that at least three out of six laminin reactive and five out of six non-laminin-binding mAbs are encoded by germline VH genes (a total of eight out of 12 mAbs). Of the eight mAbs that are encoded by germline VH genes, seven are of a non-immunoglobulin M (IgM) isotype, indicating that isotype switching has occurred in these mAbs in the absence of somatic mutations. The mutations observed in the VH genes of the four remaining mAbs do not provide strong evidence for antigenic selection. The data support the notion that B cells in this model of MGP are not subjected to affinity maturation and probably result from polyclonal B-cell activation. PMID- 11412308 TI - Detection of anaphylatoxin receptors on CD83+ dendritic cells derived from human skin. AB - Dendritic cells (DC) are recruited to sites of inflammation for the initiation of immune responses. As the anaphylatoxins C5a and C3a are important mediators of inflammation, we investigated the expression of their receptors (C3aR and C5aR) on human DC. DC were isolated from human skin or generated from purified blood monocytes and were identified by their expression of CD1a or CD83. Freshly isolated or cultured dermal CD1a+ and CD83+ DC bound anti-C5aR and anti-C3aR monoclonal antibodies (mAbs), as detected by flow cytometry. C5a induced calcium fluxes in dermal CD1a+ and CD83+ DC, which could be inhibited by C17/5, an anti C5a mAb. C3a did not induce calcium fluxes in these cells. Anaphylatoxin receptor expression was down-regulated on dermal DC by adding tumour necrosis factor-alpha (TNF-alpha) to the culture medium. On CD1a+ CD83- cells generated from isolated blood monocytes by culture with 6.25 ng/ml of granulocyte-macrophage colony stimulating factor (GM-CSF) and 125 U/ml of interleukin-4 (IL-4), expression of both C5aR and C3aR was observed. In these cells, both C5a and C3a induced calcium fluxes. After addition of TNF-alpha to the culture medium, the majority of the CD1a+ cells expressed CD83+. These cells - expressing a phenotype of 'mature DC' down-regulated the expression of the anaphylatoxin receptors and lost their reactivity to the respective ligands. Our results demonstrate the expression of the anaphylatoxin receptors C5aR and C3aR on human skin-derived DC and blood derived cells expressing the DC-associated membrane molecule, CD1a. Furthermore, the expression of anaphylatoxin receptors on CD83+ dermal DC is indicative of an intermediate stage of maturation of these cells, which was not observed on in vitro-differentiated CD83+ cells. PMID- 11412309 TI - Major histocompatibility complex class II invariant chain expression in non antigen-presenting cells. AB - In contrast to the generally accepted belief, the major histocompatibility complex (MHC) class II invariant chain (Ii) is commonly expressed intracellularly in cells that do not present exogenous antigens. Such cells include resting peripheral blood T cells and natural killer (NK) cells. In T cells, the Ii is associated with a 77 000 molecular-weight molecule (p77) that has yet to be identified. This molecule is co-precipitated with the anti-Ii monoclonal antibody (mAb) VCD-1, but not with mAb BU-45. This suggests that in the p77-Ii complex, the extracellular epitope of Ii recognized by BU-45 is hidden, whereas the Ii epitope for VCD-1 remains exposed. In antigen-presenting cells (APCs), p77 association with the Ii was minimal, if detectable. The p77-Ii association in non professional APCs suggests that the Ii may have another, more general, function other than the one accepted in antigen presentation. PMID- 11412310 TI - The role of the macrophage scavenger receptor in immune stimulation by bacterial DNA and synthetic oligonucleotides. AB - To assess the role of the macrophage scavenger receptor type A (SRA) in immune activation by CpG DNA, cytokine induction and DNA uptake were tested in vitro and in vivo using SRA knockout (SRA-/-) and wild type (WT) mice. As a source of CpG DNA, Escherichia coli DNA (EC DNA) and a 20-mer phosphorothioate oligodeoxynucleotide with two CpG motifs (CpG ODN) were used. In vitro, both EC DNA and the CpG ODN induced dose-dependent increases of interleukin (IL)-12 production by spleen cells and bone-marrow-derived macrophages (BMMPhi) from both SRA-/- and WT mice. The levels of cytokines produced by SRA-/- spleen cells and BMMPhi were similar to those of WT spleen cells and BMMPhi. When injected intravenously with CpG ODN and EC DNA, both SRA-/- and WT mice showed elevated serum levels of IL-12. To investigate further the role of the SRA, flow cytometry and confocal microscopy were performed to examine the uptake of fluorescently labelled oligonucleotides. SRA-/- and WT BMMPhi showed similarity in the extent of uptake and distribution of oligonucleotides as assessed by these two techniques. Together, these findings indicate that, while the SRA may bind DNA, this receptor is not essential for the uptake of CpG DNA or its immunostimulatory activity. PMID- 11412312 TI - The pan-chemokine inhibitor NR58-3.14.3 abolishes tumour necrosis factor-alpha accumulation and leucocyte recruitment induced by lipopolysaccharide in vivo. AB - Chemokines participate in the regulation of leucocyte recruitment in a wide variety of inflammatory processes, including host defence and diseases such as asthma, atherosclerosis and autoimmune disorders. We have previously described the properties of Peptide 3, the first broad-specificity chemokine inhibitor in vitro. Here, we report the properties of NR58-3.14.3, a retroinverso analogue of Peptide 3. NR58-3.14.3 inhibited leucocyte migration induced by a range of chemokines, including monocyte chemoattractant protein-1 (MCP-1) (2.5 nM), macrophage inflammatory protein-1alpha (MIP-1alpha) (5 nM), regulated on activation, normal T-cell expressed and presumably secreted (RANTES) (20 nM), stromal cell-derived factor-1alpha (SDF-1alpha) (25 nM) and interleukin-8 (IL-8) (30 nM), but did not affect migration induced by N-formyl-methionyl-leucyl phenylalanine (FMLP) or complement C5a (> 100 microM). NR58-3.14.3 is therefore approximately 1000-fold more potent than Peptide 3 but retains the broad-spectrum chemokine inhibitory activity of the parent peptide. In vivo, pretreatment with a systemic dose of 10 mg of NR58-3.14.3, but not the inactive derivative NR58 3.14.4, abolished leucocyte recruitment in response to intradermal injection of 500 ng of MCP-1 into rat skin. This suggests that NR58-3.14.3 is a functional chemokine inhibitor in vivo as well as in vitro. We utilized NR58-3.14.3 as a tool to investigate the role of chemokine activity during leucocyte recruitment in response to lipopolysaccharide (LPS) in vivo. NR58-3.14.3, but not NR58 3.14.4, abolished leucocyte recruitment in response to intradermal injection of 50 ng of LPS into rat skin. Furthermore, NR58-3.14.3 completely inhibited LPS induced accumulation of tumour necrosis factor-alpha (TNF-alpha). This data is consistent with a model in which multiple chemokines act in parallel upstream of TNF-alpha. NR58-3.14.3 is therefore a powerful anti-inflammatory agent in vivo, suppressing proinflammatory cytokine production and leucocyte recruitment in response to endotoxin stimulus in rat skin. PMID- 11412311 TI - Glycyrrhizin enhances interleukin-12 production in peritoneal macrophages. AB - Interleukin-12 (IL-12) is a monocyte/macrophage-derived cytokine that plays a prominent role in the development of T helper type 1 (Th1) cell-mediated immune responses. Glycyrrhizin (GL), an aqueous extract of liquorice root, used as Chinese medicine, is known to have various immunomodulating activities. In this study, GL showed a dose-dependent priming effect on lipopolysaccharide (LPS) induced IL-12 p40 and IL-12 p70 (heterodimer of p40 and p35) protein production by peritoneal macrophages (PM). The maximal effect was observed when GL was intraperitoneally administered 12 hr before the PM were harvested and stimulated in vitro with LPS. The increases in IL-12 p70 and p40 protein production were primarily due to up-regulated transcription of IL-12 p35 and p40 messenger RNAs (mRNAs), as demonstrated by RNase protection assay. The augmentation of IL-12 p40 mRNA expression induced by GL pretreatment was associated with increased NF kappaB activation. Moreover, GL exhibited the same priming effect on IL-12 production in interferon-gamma knockout (IFN-gamma-/-) mice. The production of granulocyte-macrophage colony-stimulating factor (GM-CSF) was not induced at any time point after GL pretreatment. These findings demonstrated the ability of GL to enhance LPS-induced IL-12 production by peritoneal macrophages, and indicated that the priming effect of GL on IL-12 production was independent of both IFN gamma and GM-CSF. PMID- 11412314 TI - Comparison of methods for determining the numbers and species distribution of coliform bacteria in well water samples. AB - AIMS: Enumeration of coliform bacteria and Escherichia coli is the most widely used method in the estimation of hygienic quality of drinking water. The yield of target bacteria and the species composition of different populations of coliform bacteria may depend on the method.Three methods were compared. METHODS AND RESULTS: Three membrane filtration methods were used for the enumeration of coliform bacteria in shallow well waters. The yield of confirmed coliform bacteria was highest on Differential Coliform agar, followed by LES Endo agar. Differential Coliform agar had the highest proportion of typical colonies, of which 74% were confirmed as belonging to the Enterobacteriaceae. Of the typical colonies on Lactose Tergitol 7 TTC agar, 75% were confirmed as Enterobacteriaceae, whereas 92% of typical colonies on LES Endo agar belonged to the Enterobacteriaceae. LES Endo agar yielded many Serratia strains, Lactose Tergitol 7 TTC agar yielded numerous strains of Rahnella aquatilis and Enterobacter, whereas Differential Coliform agar yielded the widest range of species. CONCLUSION: The yield of coliform bacteria varied between methods. Each method compared had a characteristic species distribution of target bacteria and a typical level of interference of non-target bacteria. Identification with routine physiological tests to distinct species was hampered by the slight differences between species. High yield and sufficient selectivity are difficult to achieve simultaneously, especially if the target group is diverse. SIGNIFICANCE AND IMPACT OF THE STUDY: The results showed that several aspects of method performance should be considered, and that the target group must be distinctly defined to enable method comparisons. PMID- 11412315 TI - Fermentations of fructo-oligosaccharides and their components by Bifidobacterium infantis ATCC 15697 on batch culture in semi-synthetic medium. AB - AIMS: To compare the physiological behaviour of Bifidobacterium infantis ATCC 15697 growing on synthetic oligofructose or its components. METHODS AND RESULTS: The studies were carried out in regulated or non-regulated batch cultures on semi synthetic media. Differences between the carbohydrate utilization patterns with glucose, fructose, sucrose and fructo-oligosaccharides (FOS) were determined. Glucose was the preferred substrate for growth and biomass production, whereas fructose was the best for lactate and acetate production. With sucrose, biomass production reached the level obtained with glucose, whereas with FOS, more metabolites were produced, as with fructose. In a mixture of FOS, the shorter saccharides were used first and fructose was released in the medium. Fructofuranosidase, an enzyme necessary to hydrolyse FOS, was inducible by fructose. CONCLUSION: Glucose contained in FOS and sucrose might sustain growth and cell production, while fructose might enable the production of major metabolites. SIGNIFICANCE AND IMPACT OF THE STUDY: A better understanding of the bifidogenic nature of oligofructose has been gained. PMID- 11412316 TI - Assessment of toxic potential of local Jordanian Bacillus thuringiensis strains on Drosophila melanogaster and Culex sp. (Diptera). AB - AIMS: To assess the toxic potential of different local Jordanian Bacillus thuringiensis isolates on larvae of Drosophila melanogaster and Culex sp. METHODS AND RESULTS: Scanning electron microscopy revealed the presence of spherical, bi pyramidal, and bi-pyramidal and cuboidal parasporal bodies produced by the toxic isolates. Spherical inclusions dominated. The toxicity of the isolates to the two insects, determined using 24-well plates or vials, indicated that the 50% lethal concentration (LC50) of the bacterial suspension for D. melanogaster and Culex sp. larvae varied from 4.60 to 8.65, and from 5.30 to 6.74, respectively. CONCLUSION: Comparison of the LC50 values of isolate 82 with those of the reference strain B. t. israelensis showed that this isolate has a higher toxicity potential. SIGNIFICANCE AND IMPACT OF THE STUDY: Some local Jordanian B. thuringiensis isolates exhibit toxic potential that could be used to control some important pests, and could replace chemical pesticides. PMID- 11412317 TI - Isolation and characterization of human colonic bacteria able to hydrolyse chlorogenic acid. AB - AIMS: Conjugated hydroxycinnamates, such as chlorogenic acid (caffeoyl-quinic acid), are widely consumed in a Western diet, coffee being one of the richest sources. Ingested hydroxycinnamate esters can reach the large intestine essentially unaltered, and may then be hydrolysed by esterases produced by the indigenous microflora. This study is aimed at identifying bacterial species responsible for the release of natural antioxidants, such as hydroxycinnamic acids, in the human large intestine. METHODS AND RESULTS: Thirty-five isolates recovered after anaerobic batch culture incubation of human faecal bacteria in a chlorogenic acid-based medium were screened for cinnamoyl esterase activity. Six isolates released the hydroxycinnamate, ferulic acid, from its ethyl ester in a plate-screening assay, and these were identified through genotypic characterization (16S rRNA sequencing) as Escherichia coli (three isolates), Bifidobacterium lactis and Lactobacillus gasseri (two strains). Chlorogenic acid hydrolysing activities were essentially intracellular. These cinnamoyl esterase producing organisms were devoid of other phenolic-degrading activities. CONCLUSION: The results show that certain gut bacteria, including some already recognized as potentially health-promoting (i.e. species belonging to the genera Bifidobacterium and Lactobacillus), are involved in the release of bioactive hydroxycinnamic acids in the human colon. SIGNIFICANCE AND IMPACT OF THE STUDY: Free hydroxycinnamates, including caffeic, ferulic and p-coumaric acids, exhibit antioxidant and anticarcinogenic properties both in vitro and in animal models. Given that the gut flora has a major role in human nutrition and health, some of the beneficial effects of phenolic acids may be ascribed to the microflora involved in metabolism. PMID- 11412318 TI - Evolution of microbial populations and biogenic amine production in dry sausages produced in Southern Italy. AB - AIMS: To evaluate the occurrence and evolution of biogenic amines during ripening of fermented sausages and their relationship with physico-chemical and microbiological properties of the product. METHODS AND RESULTS: Salsiccia and Soppressata were obtained from artisanal and industrial plants in Basilicata and pH, aW, microbial counts and biogenic amine content were measured. A high variability in amine content was observed. 2-Phenylethylamine and histamine were rarely found, while the tyramine, putrescine and cadaverine content increased during ripening. No correlation was found between individual biogenic amine content, microbial counts or physico-chemical parameters. CONCLUSION: Starter cultures did not necessarily prevent the production of biogenic amines whose total contents were usually higher in Soppressata, a product with a larger diameter and aW compared with Salsiccia. SIGNIFICANCE AND IMPACT OF THE STUDY: Literature findings on biogenic amine content and the evolution of microbial populations were confirmed. Normal ranges for amine content in Salsiccia and Soppressata are reported. PMID- 11412319 TI - Potential occurrence of adhering living Bacillus spores in milk product processing lines. AB - AIMS: The hygienic risk associated with microbial soil on surfaces of milk processing lines was evaluated, based on experimental results. METHODS AND RESULTS: From a panel of Bacillus spores isolated from milk products, B. cereus CUETM 98/4, was found to be highly resistant to heat (D100=3.32 min in whole milk) and oxidant disinfectant (70% lethality of adherent spores with Ikalin 2%). From adhesion trials, up to 1.1 x 10(7) spores cm(-2) were found to be adherent to solid surfaces when suspended in saline or in custard (10(5) and 10(7) cfu ml( 1)), and over 10% of these adherent spores would resist the cleaning procedure. CONCLUSION: A highly contaminated milk (10(5) cfu ml(-1)) subjected to a current sterilization process (8 log reduction) led to a residual contamination of less than 1 cfu in the representative processing line after a complete production run. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlighted the fact that under appropriate processing conditions (efficient sterilization and cleaning procedures), even disinfection would be sufficient to eliminate any contamination risk. Conversely, the disinfection procedure becomes an essential step under inappropriate processing conditions. PMID- 11412320 TI - Factors influencing attachment of thermophilic bacilli to stainless steel. AB - AIMS: This project aimed to investigate the mechanism of attachment of the vegetative cells and spores of thermophilic bacilli to stainless steel with a view to devising strategies to limit biofilm development and survival. METHODS AND RESULTS: Spores and vegetative cells of bacterial isolates were exposed to protein denaturing agents (sodium dodecyl sulphate (SDS) and trypsin) and polysaccharide removing agents (sodium metaperiodate, trichloroacetic acid (TCA) and lysozyme). Treatment with sodium metaperiodate, TCA and lysozyme increased the number of vegetative cells attaching in many of the strains studied, while SDS and trypsin decreased attachment. Spores attached to stainless steel in greater numbers than vegetative cells, and the various treatments had less effect on this attachment than for vegetative cells. Viability of the cells or spores was not an important factor in attachment, as cells and spores rendered non viable also attached to stainless steel in similar numbers. Coating the stainless steel with skim milk proteins decreased the attachment of both vegetative cells and spores. There was no correlation between the degree of attachment and the amount of extracellular polysaccharide (EPS) produced by each strain, surface hydrophobicity or zeta potential of vegetative cells or spores, though spores were found to be more hydrophobic than vegetative cells. CONCLUSIONS: The results suggest that biofilm formation by these thermophilic bacilli is probably a multifactorial process, and that cell-surface proteins play a very important role in the initial process of attachment during the formation of biofilms by these bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: This information will provide direction for developing improved cleaning systems to control biofilms of thermophilic bacilli in dairy manufacturing plants. PMID- 11412321 TI - Differentiation of Lactobacillus delbrueckii subspecies by ribotyping and amplified ribosomal DNA restriction analysis (ARDRA). AB - AIMS: To differentiate the subspecies of Lactobacillus delbrueckii, subsp. delbrueckii, subsp. lactis and subsp. bulgaricus. METHODS AND RESULTS: Amplified ribosomal DNA restriction analysis (ARDRA) and ribotyping were applied to over 30 strains. Both methods analyse the ribosomal genes which carry useful information about the evolutionary and taxonomic relationship among bacteria. The methods proved to be reliable and highly reproducible. ARDRA was applied to 16S rDNA, 23S rDNA and the IGS region, thus covering the whole rrn operon with eight restriction enzymes. Only EcoRI differentiated Lact. delbrueckii subsp. bulgaricus from Lact. delbrueckii subsp. delbrueckii/Lact. delbrueckii subsp. lactis, which confirmed the finding of other authors. Ribotyping with different enzymes under precisely optimized conditions revealed a high level of strain polymorphism. Only ribotyping with EcoRI allowed differentiation of the three subspecies on the basis of typical hybridization patterns. CONCLUSION: The successful differentiation of the three subspecies of Lact. delbrueckii by EcoRI ribotyping offers a new possibility for precise identification and differentiation of strains and new isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: Both methods could be used for differentiation of Lact. delbrueckii subspecies. PMID- 11412322 TI - Characterization of Aeromonas and Vibrio species isolated from a drinking water reservoir. AB - AIMS: To study the phenotypic and chemotaxonomic (i.e. phospholipid and cellular fatty acid composition) characteristics of environmental Aeromonas spp. and Vibrio spp. isolated from a drinking water reservoir near Vladivostok City, and the application of some chemotaxonomic markers for discrimination of the two genera and species. METHODS AND RESULTS: Presumptive Aeromonas species were dominant in surface water samples (up to 25% of the total number of bacteria recovered). These strains were consistent with respect to the cultural and biochemical properties used to define the species Aeromonas sobria (seven strains) and Aer. popoffii (three strains). Vibrio mimicus (two strains) and Vibrio metschnikovii (one strain) were identified according to phenotypic features and cellular fatty acid composition. CONCLUSION: Environmental Aer. sobria isolates were atypical in their ability to grow at 42 degrees C, and were haemolytic, proteolytic and cytotoxic. Although it was present in a high proportion in the water samples, atypical Aer. sobria is not an indicator of polluted water. SIGNIFICANCE AND IMPACT OF THE STUDY: The incidence of Aeromonas in the drinking water reservoirs in the Far East of Russia is reported for the first time. PMID- 11412323 TI - Characterization of lactic acid bacteria strains on the basis of neutral volatile compounds produced in whey. AB - AIMS: Seventy-eight strains of lactic acid bacteria belonging to five genera and showing six different phenotype combinations of Lac (lactose fermentation), Prt (proteolytic activity) and Cit (citrate degradation) characters were investigated for their main flavouring properties with the aim to detect variability among and within the groups. METHODS AND RESULTS: High resolution gas chromatography-mass spectrometry analysis of neutral volatile compounds produced in whey showed that, considering both neo-formation compounds and substances quantified in the whey cultures at different concentrations in comparison to the extract from sterile whey, the groups of lactococci, enterococci, thermophilic streptococci and mesophilic lactobacilli produced a higher number of volatiles than thermophilic lactobacilli and leuconostocs. Applying principal component analysis (PCA) to the results, enterococci, mesophilic lactobacilli and thermophilic streptococci showed a broad diversity, while lactococci included rather similar strains as well as strains with special flavouring properties. Applying PCA to thermophilic streptococci and enterococci, to lactococci and enterococci, to lactococci and thermophilic streptococci, or to mesophilic and thermophilic lactobacilli, the strains gathered consistently with their systematic position. CONCLUSION: The study evidenced strains producing some volatile compounds responsible for food flavouring. Flavouring properties were variable among the systematic groups and in some cases different within the same bacterial group. SIGNIFICANCE AND IMPACT OF THE STUDY: The potential of the findings is discussed with reference to the development of flavouring adjuncts for the dairy industry. PMID- 11412324 TI - Phenotypic diversity of lactic acid bacteria isolated from fermented sausages produced in Basilicata (Southern Italy). AB - AIMS: to evaluate the evolution of lactic acid bacteria (LAB) populations in traditional fermented sausages (salsiccia and soppressata) produced in artisanal and industrial plants in Basilicata (Southern Italy). METHODS AND RESULTS: Four hundred and fourteen lactic acid bacteria (LAB) cultures were isolated from samples of sausages at different stages of ripening. A phenotypic characterization of the isolates was carried out using a set of 28 tests, and 34 clusters were identified at the 80% similarity level using hierarchical cluster analysis. Of the isolates 50% were identified as Lactobacillus sakei (with several biotypes), 22% as Pediococcus spp. (mainly Ped. pentosaceus), 7% as Leuconostoc (Leuc. carnosum, Leuc. gelidum, Leuc. pseudomesenteroides), 6% as Lact. plantarum, 1% as Lact. curvatus. Other lactobacilli, including unidentified species, were present in lower numbers. CONCLUSION: The phenotypic diversity and composition of the LAB flora varied as a function of the production plant, product type and ripening time. SIGNIFICANCE AND IMPACT OT THE STUDY: A new procedure based on bootstrapping and Multidimensional Scaling was successfully used to obtain a graphical representation of the evolution of the LAB populations. PMID- 11412325 TI - The impact of flow rate (simulated leaching) on plasmid transfer frequency between bacteria in a model rhizosphere system. AB - AIMS: To investigate the effect of flow rate and inoculation order on plasmid transfer frequency between bacteria in a model rhizosphere system. METHODS AND RESULTS: A physical model system was constructed and used to demonstrate that although flow rate did affect plasmid transfer frequency for an introduced strain, the flow rates necessary for a significant effect on an established population were much higher than typical water flow rates found through soil. Plasmid transfer frequency was highly sensitive to strain inoculation order. CONCLUSION: Flow rate may not have a significant effect on plasmid transfer frequency between established bacterial populations in the rhizosphere. SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributes to the current debate over the release and spread of genetically modified organisms into the environment. It also demonstrates that model controlled systems may be used to rapidly obtain initial data about the potential behaviour of microorganisms, prior to more costly and lengthy glasshouse and field trials. PMID- 11412326 TI - Transfer of bacteria from fabrics to hands and other fabrics: development and application of a quantitative method using Staphylococcus aureus as a model. AB - AIMS: To develop and apply a quantitative protocol for assessing the transfer of bacteria from bleached and undyed fabrics of 100% cotton and 50% cotton + 50% polyester (poly cotton) to fingerpads or other pieces of fabric. METHODS AND RESULTS: Test pieces of the fabrics were mounted on custom-made stainless steel carriers to give a surface area of 1 cm in diameter, and each piece seeded with about 10(5) cfu of Staphylococcus aureus from an overnight broth culture; the inoculum contained 5% fetal bovine serum as the soil load. Transfer from fabric to fabric was performed by direct contact using moist and dry fabrics. Transfers from fabrics to fingerpads of adult volunteers were tested using moist, dry and re-moistened pieces of the fabrics, with or without friction during the contact. Bacterial transfer from fabrics to moistened fingerpads was also studied. All the transfers were conducted under ambient conditions at an applied pressure of 0.2 kg cm(-2). After the transfer, the recipient fingerpads or fabric pieces were eluted, the eluates spread-plated, along with appropriate controls, on tryptic soy agar and the percentage transfer calculated after the incubation of the plates for 24 h at 37 degrees C. CONCLUSION: Bacterial transfer from moist donor fabrics using recipients with moisture was always higher than that to and from dry ones. Friction increased the level of transfer from fabrics to fingerpads by as much as fivefold. Bacterial transfer from poly cotton was consistently higher when compared with that from all-cotton material. SIGNIFICANCE AND IMPACT OF THE STUDY: The data generated should help in the development of better models to assess the role fabrics may play as vehicles for infectious agents. Also, the basic design of the reported methodology lends itself to work with other types of human pathogens. PMID- 11412327 TI - Lactobacillus casei and Lactobacillus plantarum initiate catabolism of methionine by transamination. AB - AIMS: To study the ability of Lactobacillus casei and Lact. plantarum strains to convert methonine to cheese flavour compounds. METHODS AND RESULTS: Strains were assayed for methionine aminotransferase and lyase activities, and amino acid decarboxylase activity. About 25% of the strains assayed showed methionine aminotransferase activity. The presence of glucose in the reaction mixture increased conversion of methionine to 4-methylthio-2-ketobutanoate (KMBA) and 4 methylthio-2-hydroxybutanoate (HMBA) in all strains. The methionine aminotransferase activity in Lact. plantarum and Lact. casei showed variable specificity for the amino group acceptors glyoxylate, ketoglutarate, oxaloacetate and pyruvate. None of the strains showed methionine lyase or glutamate and methionine decarboxylase activities. CONCLUSION: The presence of amino acid converting enzymes in lactobacilli is strain specific. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings of this work suggest that lactobacilli can be used as adjuncts for flavour formation in cheese manufacture. PMID- 11412328 TI - A strain of Pseudomonas fluorescens with two lipase-encoding genes, one of which possibly encodes cytoplasmic lipolytic activity. AB - AIMS: A lipase-encoding gene (lipA) from a psychrotrophic strain of Pseudomonas fluorescens C9 has previously been characterized. It was also shown that when this gene was insertionally-inactivated, lipase activity was retained, suggesting that a second lipase may be present in this strain. The aim of this study was to determine whether this was the case. METHODS AND RESULTS: Using molecular cloning, chromosomal mutagenesis and enzymatic analysis, the presence of a second lipase-encoding gene (lipB) has been confirmed. The molecular weights of the putative products of lipA and lipB are 33 and 64.5 kDa, respectively, and their sequences are quite dissimilar (< 10% sequence identity). The lipB gene encodes a secreted lipase and is solely responsible for the 'lipolytic phenotype' of Ps. fluorescens C9. Expression of the lipA gene can be detected when expressed using an expression vector, but activity was only detected intracellularly in Ps. fluorescens C9, and not in the culture medium. CONCLUSION: Pseudomonas fluorescens C9 contains two dissimilar lipases. One (LipB) is secreted and responsible for the lipolytic phenotype; the evidence suggests that the other (LipA) could be intracellular, but it could be secreted and not detectable. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacteria may contain more than one lipase activity. Ascribing phenotypes to particular enzymes therefore requires mutational analysis. The notion of an intracellular lipase activity is novel, and, if further substantiated, begs the question as to its normal substrate and physiological role. PMID- 11412329 TI - A simple chemiluminescence-based method for rapid enumeration of Listeria spp. microcolonies. AB - AIMS: Listeria monocytogenes is capable, under certain conditions, of producing chemiluminescence which is amplified by luminol. This property was used to detect and count microcolonies of Listeria spp. in a few hours, without the use of a microscope. METHODS AND RESULTS: After trapping Listeria cells on polyvinylidene fluoride membranes, a chemiluminescence mixture was sprayed onto the membrane. The chemiluminescent spots emitted were analysed by a charge-coupled device camera connected to a data-processing system, which restored the intensity of the signals into three dimensional images. The intensity of the luminescence of microcolonies was improved by addition of cellobiose, and by brief exposure to u.v. light. CONCLUSION: Microcolonies of Listeria spp. can be imaged and counted by luminol-enhanced chemiluminescence with a photon-counting system. SIGNIFICANCE AND IMPACT OF THE STUDY: This method can be applied to the rapid detection and counting of Listeria spp. in raw milk. PMID- 11412330 TI - Comparison of selective and non-selective enrichment media in the detection of Listeria monocytogenes from meat containing Listeria innocua. AB - AIMS: This study investigated whether the higher incidence of recovery from meat of Listeria innocua compared with L. monocytogenes could be due to the laboratory media used, leading to an artificially lower detection of the pathogenic species, L. monocytogenes. METHODS AND RESULTS: Minced beef was inoculated with L. monocytogenes, L. innocua, or a mixture of these species, and stored at 0 or 10 degrees C under vacuum or aerobic conditions for up to 28 days. Listeria were recovered from the minced beef using selective (University of Vermont Medium, UVM) and non-selective (Buffered Peptone Water, BPW) enrichment broths after 0, 14, and 28 days of storage. In general, there were no significant differences (P < 0.05) between the numbers of L. monocytogenes recovered from minced beef samples after 24 h enrichment in BPW and the numbers recovered using UVM. In addition, the presence of L. innocua in meat samples containing L. monocytogenes did not significantly (P < 0.05) affect the numbers of L. monocytogenes recovered using either enrichment broth. In most cases there were no significant differences (P < 0.05) between the numbers of L. innocua recovered from minced beef samples after 24 h enrichment in BPW compared with numbers recovered using UVM. CONCLUSION: Listeria innocua was found to have no significant competitive advantage over L. monocytogenes in selective or non-selective enrichment media. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that, in some instances, the use of a selective enrichment broth offers no advantage over a non selective enrichment broth for the recovery of Listeria species from minced beef. PMID- 11412331 TI - Listeria monocytogenes adheres to many materials found in food-processing environments. AB - AIMS: To investigate the adhesion of Listeria monocytogenes 10403S to 17 different, food-use approved materials representing metals, rubbers and polymers. METHODS AND RESULTS: Adhesion assays were conducted by placing 'coupons' of the materials in planktonic cultures at 30 degrees C, and then immediately withdrawing them ('short contact') or leaving them submerged in the cultures for 2 h. Adherent cells were recovered by sonication. In the short contact experiments, the logarithm of the mean viable counts ranged from 3.67 +/- 0.43 to 4.78 +/- 0.38. After 2 h contact time, the numbers of adherent cells had increased significantly for all materials with the exception of polypropylene. The highest count (6.33 +/- 0.31) recorded was for stainless steel 405. CONCLUSION: Adhesion to a wide range of materials was time-dependent and characterized by reversible and irreversible stages. SIGNIFICANCE AND IMPACT OF THE STUDY: Adhesion test protocols must account for cell carry-over and cells which are only weakly bound. Material selection may only have a limited role in reducing food contamination by listeria. PMID- 11412332 TI - Separation and characterization of the 1,3-propanediol and glycerol dehydrogenase activities from Clostridium butyricum E5 wild-type and mutant D. AB - AIMS: Clostridium butyricum E5 wild-type and mutant E5-MD were cultivated in chemostat culture on glycerol in order to compare the properties of two key enzymes of glycerol catabolism, i.e. propanediol and glycerol dehydrogenase. METHODS AND RESULTS: These two enzymes, which belong to the dha regulon, were separated by gel filtration. Both dehydrogenase activities displayed similar properties, such as pH optimum values, specificity towards physiological substrates and dependence on Mn2+. Both strains accumulate glycerol at high levels. CONCLUSION: The mutant D strain contained a propanediol dehydrogenase activity which had a low affinity for its physiological substrate, leading to the conclusion that this strain would seem more resistant to the toxic effect of 3 hydroxypropionaldehyde than the wild-type. SIGNIFICANCE AND IMPACT OF THE STUDY: These properties make Cl. butyricum mutant D strain the best candidate so far to be used as a biotechnological agent for the bioconversion of glycerol to 1,3 propanediol. PMID- 11412333 TI - An in vitro study of the pH-lowering potential of salivary lactobacilli associated with dental caries. AB - AIMS: Lactobacilli are known to produce acids from sucrose or glucose. This acid production can cause a drop in pH which is sufficiently significant to demineralize the hard tissues of the teeth. Some authors have demonstrated the benefits of substituting sorbitol or xylitol for sucrose. The aim of this work was to study the acid production of salivary lactobacilli with one test sugar (glucose) and two polyols (sorbitol and xylitol). METHODS AND RESULTS: The pH lowering potential of three strains of oral lactobacilli was recorded with glucose or one of the polyols at three different concentrations. The results showed that polyols were broken down by certain strains of lactobacilli. When this degradation took place, the pH dropped to values sufficiently low to demineralize the hard tissues of the teeth. CONCLUSIONS: Further studies must be carried out on the metabolism of polyols before encouraging their widespread substitution for sucrose. PMID- 11412334 TI - Hypothalamic transforming growth factor beta1 and basic fibroblast growth factor mRNA expression is modified during the rat oestrous cycle. AB - The present observations show that the mRNA levels of two growth factors, previously described to be involved in the control of neurones synthesizing the luteinizing hormone releasing hormone (LHRH) [i.e. transforming growth factor beta1 (TGFbeta1) and basic fibroblast growth factor (bFGF)], fluctuate in the hypothalamus of adult female rats during the oestrous cycle. In particular, the expression of TGFbeta1-mRNA shows a peak on the morning of the day of proestrus, which precedes the increased secretion of the two gonadotrophins that occurs on that day. In the case of bFGF, the peak is evident in the evening of the same day and is concomitant with that of the gonadotrophins. We evaluated the effects of ovariectomy and of exogenous oestrogens on the mRNA levels of these two growth factors in the hypothalamus. The data indicate that 3 weeks of ovariectomy are not able to change the hypothalamic messenger levels of the two growth factors considered, which remain at the levels found in diestrus 1, and that 17beta oestradiol is able to induce a significant increase of both TGFbeta1- and of bFGF mRNA levels in the hypothalamus of the ovariectomized rat. The present in vivo observations support the concept, previously proposed on the basis of in vitro data, that growth factors, such as TGFbeta1 and bFGF, play a role in the hypothalamic control of reproduction, and suggest that the control of LHRH dynamics involves a strict cooperation between gonadal steroids and growth factors. PMID- 11412335 TI - Anorexic but not pyrogenic actions of interleukin-1 are modulated by central melanocortin-3/4 receptors in the rat. AB - The cytokine interleukin-1 (IL-1), which mediates many responses to infection and injury, induces anorexia and fever through direct actions in the central nervous system. The melanocortin neuropeptides, such as alpha melanocyte-stimulating hormone (alpha-MSH), reportedly antagonize many actions of IL-1, including fever and anorexia. However, it is unknown whether endogenous melanocortins modulate anorexia induced by IL-1. The objective of the present study was to establish the effect of endogenous melanocortins on IL-1-induced anorexia and fever in the rat. Intracerebroventricular (i.c.v.) injection of IL-1beta caused a significant reduction in food intake and body weight gain, and a rise in core body temperature in conscious rats. Coadministration of the melanocortin-3/4 receptor (MC3/4-R) antagonist, SHU9119, reversed IL-1beta-induced reductions in food intake and body weight, but did not affect the febrile response to IL-1beta. These data suggest IL-1beta may elicit its effects on food intake through the melanocortin system, predominantly via the MC3-R or MC4-R. In contrast, IL-1beta induced fever does not appear to be mediated or modulated by MC3-R or MC4-R activity. PMID- 11412336 TI - Skeletal growth acceleration with growth hormone secretagogues in transgenic growth retarded rats: pattern-dependent effects and mechanisms of desensitization. AB - The transgenic growth retarded (Tgr) rat is the first genetic model of growth hormone (GH) deficiency whose growth can be accelerated with exogenous GH secretagogues (GHSs). In this study, we have demonstrated that GHS-receptor (GHS R) mRNA expression in the arcuate nucleus of Tgr rats was not significantly different to that in wild-type littermates. We have confirmed that GHS-induced elevation in body weight gain was accompanied by acceleration of skeletal growth, and that the effects of the GHS, GHRP-6, were both dose- and pattern-dependent. The growth response with continuous infusion of GHRP-6 was transient, accompanied by suppression of GH and corticosterone responses to bolus injection of GHRP-6. This desensitization occurred without downregulation of arcuate GHS-R mRNA expression, but was accompanied by elevated periventricular somatostatin mRNA expression. In contrast, pulsatile (3-hourly) infusion of GHRP-6 produced sustained growth and GH responses, which were accompanied by suppression of corticosterone responses and elevated arcuate GH-releasing factor (GRF) mRNA expression. Skeletal growth was further accelerated by coinfusion of GRF, but significant depletion of pituitary GH stores suggested that this growth rate may not be sustainable. These experiments confirm the importance of the Tgr rat for investigating the growth promoting potential of the GHSs in the context of GH deficient dwarfism, and suggest that elevated somatostatin expression may mediate the suppression of the GRF-GH and hypothalamo-pituitary-adrenal axes following continuous GHRP-6 treatment. PMID- 11412337 TI - GABAergic nature of hypothalamic leptin target neurones in the ventromedial arcuate nucleus. AB - Leptin is an adipose tissue-derived cytokine hormone, which reduces body weight via interactions with hypothalamic neurones. Leptin receptors capable of activating the JAK-STAT signal transduction pathway are expressed at high levels in the hypothalamus, particularly in the arcuate nucleus. In order to identify the chemical mediators of leptin's action in the hypothalamus, we have examined whether GABA neurones of the hypothalamic arcuate nucleus contain leptin receptors and the leptin-activated transcription factor STAT3. GABAergic neurones, as visualized by antisera to the GABA-synthesizing enzyme glutamic acid decarboxylase (GAD) and GABA, were demonstrated in the ventromedial and ventrolateral parts of the arcuate nucleus. GABA neurones in the ventromedial arcuate nucleus were shown to contain leptin receptor immunoreactivity, as revealed using an antiserum generated to a sequence common to all isoforms of the leptin receptor (Ob-R), as well as an antiserum generated to the carboxy-terminal end of the long leptin receptor (Ob-Rb), and immunoreactivity for the leptin induced signal transduction molecule STAT3. Ventromedial GABA neurones were also shown to contain neuropeptide Y, whereas ventrolateral proopiomelanocortin containing neurones lacked GAD and GABA immunoreactivity. Levels of mRNA for GAD65, GAD67 and the vesicular GABA transporter (VGAT) were analysed in the arcuate nucleus of leptin-deficient ob/ob mice and lean control mice by in situ hybridization. No significant differences in GAD65, GAD67 or VGAT mRNA were detected in the arcuate nucleus of ob/ob mice as compared to lean control mice. The presence of leptin receptor and STAT3 in GABAergic arcuate neurones, but absence of changes in gene transcription for GAD and VGAT mRNA suggests, that leptin does not transcriptionally regulate the expression of proteins involved in GABAergic transmission in arcuate neurones. However, mechanisms other than transcriptional regulation for leptin to influence arcuate GABA neurones may exist. PMID- 11412338 TI - Angiotensin II induces inward currents in subfornical organ neurones of rats. AB - The action of angiotensin II on subfornical organ (SFO) neurones was studied using whole-cell current and voltage-clamp recordings in rat slice preparations. In the current-clamp mode, membrane depolarization in response to angiotensin II was accompanied by an increased frequency of action potentials and an increased membrane conductance. In the voltage-clamp mode, angiotensin II elicited inward currents in a dose-dependent manner. The net angiotensin II-induced inward currents were voltage-independent, with a mean reversal potential of -29.8 +/- 6.2 mV. Amplitudes of the angiotensin II-induced inward currents were decreased during perfusion with a low sodium medium. The angiotensin II-induced inward currents were blocked by the AT1 antagonist losartan, and were partially blocked by the AT2 antagonist PD-123319. Neurones which were sensitive to angiotensin II were found in the peripheral region of the SFO, whereas neurones in the central region were less sensitive to angiotensin II. These results suggest that angiotensin II induces inward currents, with opening of nonselective cation channels through mainly AT1 receptors in a subpopulation of SFO neurones of rats. PMID- 11412339 TI - The insulin hypoglycemia test: hypoglycemic criteria and reproducibility. AB - The insulin hypoglycemia test (IHT) is widely regarded as the "gold standard" for dynamic stimulation of the hypothalamic-pituitary-adrenal (HPA) axis. This study aimed to investigate the temporal relationship between a rapid decrease in plasma glucose and the corresponding rise in plasma adenocorticotropic hormone (ACTH), and to assess the reproducibility of hormone responses to hypoglycemia in normal humans. Ten normal subjects underwent IHTs, using an insulin dose of 0.15 U/kg. Of these, eight had a second IHT (IHT2) and three went on to a third test (IHT3). Plasma ACTH and cortisol were measured at 15-min intervals and, additionally, in four IHT2s and the three IHT3s, ACTH was measured at 2.5- or 5-min intervals. Mean glucose nadirs and mean ACTH and cortisol responses were not significantly different between IHT1, IHT2 and IHT3. Combined data from all 21 tests showed the magnitude of the cortisol responses, but not the ACTH responses, correlated significantly with the depth and duration of hypoglycemia. All subjects achieved glucose concentrations of of < or = 1.6 mmol/l before any detectable rise in ACTH occurred. In the seven tests performed with frequent sampling, an ACTH rise never preceded the glucose nadir, but occurred at the nadir, or up to 15 min after. On repeat testing, peak ACTH levels varied markedly within individuals, whereas peak cortisol levels were more reproducible (mean coefficient of variation 7%). In conclusion, hypoglycemia of < or = 1.6 mmol/l was sufficient to cause stimulation of the HPA axis in all 21 IHTs conducted in normal subjects. Nonetheless, our data cannot reveal whether higher glucose nadirs would stimulate increased HPA axis activity in all subjects. Overall, the cortisol response to hypoglycemia is more reproducible than the ACTH response but, in an individual subject, the difference in peak cortisol between two IHTs may exceed 100 nmol/l. PMID- 11412340 TI - Analysis of steroid-induced genes in the rat preoptic area-anterior hypothalamus using a differential-display reverse transcriptase-polymerase chain reaction. AB - Steroid hormones modulate a variety of physiological functions in the hypothalamus. We attempted to identify steroid-regulated genes in the rat preoptic area-anterior hypothalamus by comparing differentially expressed mRNAs. Adult female rats were ovariectomized and, 1 week later, a silastic capsule containing 17beta-oestradiol (180 microg/ml) was subcutaneously implanted. After 2 days, a single injection of progesterone (1 mg) was administered at 10.00 h and rats were killed at 17.00 h on the same day. Differential-display polymerase chain reaction followed by Northern blot analysis showed that 10 clones were differentially regulated. Using homology search in Genbank, three genes were identified as sodium, potassium-ATPase beta1, protein kinase C-binding Nell homologue protein and evectin-1. Further characterization of 10 clones showed that the expression patterns were tissue-specific and differentially regulated during puberty. Among these, mRNAs for protein kinase C-binding Nell-homologue protein, evectin-1 and human CGI-118 protein-like gene were induced after vagina opening, and differentially expressed during the oestrous cycle. Taken together, several steroid-regulated genes identified in the present study may play an important role in regulating hypothalamic functions, including puberty and the oestrous cycle. PMID- 11412341 TI - Pacap stimulation of gonadotropin-II secretion in goldfish pituitary cells: mechanisms of action and interaction with gonadotropin releasing hormone signalling. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) has recently been shown to be a hypophysiotropic factor in the goldfish. In this study, we examined the mechanisms of PACAP action on goldfish maturational gonadotropin (GTH-II) release using primary cultures of pituitary cells. The GTH-II response to mammalian PACAP1-38 (mPACAP) was inhibited by a PACAP receptor antagonist suggesting a receptor-mediated action. Addition of either an adenylate cyclase inhibitor or a protein kinase A (PKA) inhibitor reduced the mPACAP-induced GTH-II release. In addition, when GTH-II release was already stimulated by either forskolin or 8-bromo-cAMP (8Br-cAMP), mPACAP did not further increase GTH-II secretion. These results strongly implicated the involvement of an adenylate cyclase/cAMP/PKA pathway in PACAP-stimulated GTH-II release. Although mPACAP induced a rise in intracellular Ca2+ level in identified gonadotropes, results with voltage-sensitive Ca2+ channel inhibitors indicated that the GTH-II responses to mPACAP, forskolin and 8Br-cAMP did not depend upon Ca2+ entry through these channels. Two protein kinase C (PKC) inhibitors did not affect mPACAP-elicited GTH-II release, and mPACAP further increased GTH-II secretion in the presence of PKC activators. These results indicate that PKC-dependent elements are not essential for the stimulatory action of mPACAP in gonadotropes. Interestingly, while GTH-II responses to a stimulatory concentration of mPACAP were additive to responses elicited by maximal effective concentrations of two endogenous gonadotropin releasing hormones (GnRHs), a subthreshold concentration of mPACAP potentiated GnRH and PKC activator stimulation of GTH-II secretion. Similarly, submaximal concentrations of forskolin potentiated the GTH-II response to the PKC activator, tetradecanoyl phorbol acetate. These data suggest that PACAP and its cAMP-dependent signalling mechanisms provide an alternate stimulatory input to goldfish gonadotropes and may influence the effectiveness of the major neuroendocrine control exerted by the PKC-dependent GnRH signalling pathway. PMID- 11412342 TI - Maternal odours induce Fos in the main but not the accessory olfactory bulbs of neonatal male and female ferrets. AB - Previous research demonstrated that exposing gonadectomized adult ferrets to odours in oestrous female bedding induced nuclear Fos-immunoreactivity (Fos-IR; a marker of neuronal activity) in the main as opposed to the accessory olfactory system in a sexually dimorphic fashion, which was further augmented in both sexes by treatment with testosterone propionate. Ferrets are born in an altricial state and presumably use maternal odour cues to locate the nipples until the eyes open after postnatal (P) day 23. We investigated whether maternal odours augment neuronal Fos preferentially in the main versus accessory olfactory system of neonatal male and female ferret kits. Circulating testosterone levels peak in male ferrets on postnatal day P15, and mothers provide maximal anogenital stimulation (AGS) to males at this same age. Therefore, we assessed the ability of maternal odours to augment Fos-IR in the accessory olfactory bulb (AOB), the main olfactory bulb (MOB) and other forebrain regions of male and female ferret kits on P15 and investigated whether artificial AGS (provided with a paintbrush) would further enhance any effects of maternal odours. After separation from their mothers for 4 h, groups of male and female kits that were placed for 1.5 h with their anaesthetized mother had significantly more Fos-IR cells in the MOB granule cell layer and in the anterior-cortical amygdala, but not in the AOB cell layer, compared to control kits that were left on the heating pad. Artificial AGS failed to amplify these effects of maternal odours. Maternal odours (with or without concurrent AGS) failed to augment neuronal Fos-IR in medial amygdaloid and hypothalamic regions that are activated in adult ferrets by social odours. In neonatal ferrets of both sexes, as in adults, socially relevant odours are detected by the main olfactory epithelium and initially processed by the MOB and the anterior-cortical amygdala. In neonates, unlike adults, medial amygdaloid and hypothalamic neurones either do not respond to these inputs or respond in a manner that fails to induce Fos expression. PMID- 11412343 TI - The central effects of orexin-A in the hypothalamic-pituitary-adrenal axis in vivo and in vitro in male rats. AB - Orexin-A is synthesized in the posterolateral hypothalamus and immunoreactive fibres project to many central nervous system structures, including the paraventricular nucleus, which is rich in corticotropin releasing factor (CRF) neurones and neuropeptide Y (NPY) innervation. We investigated the central effects of orexin-A on the hypothalamic-pituitary-adrenal (HPA) axis by measuring plasma concentrations of corticosterone and adrenocorticotropic hormone (ACTH) in vivo. We explored the potential neuropeptide pathways involved by investigating the effects of orexin-A on CRF, NPY, arginine vasopressin (AVP) and noradrenaline release from hypothalamic explants in vitro. Intracerebroventricular (i.c.v.) injection of orexin-A (3 nmol) in male rats stimulated increases in plasma concentrations of corticosterone between 10 and 40 min after injection, and of plasma ACTH at 20 and 90 min after injection. Orexin-A significantly stimulated CRF and NPY release from hypothalamic explants in vitro. Orexin-A did not stimulate CRF release in the presence of the selective NPY Y1 receptor antagonist, BIBP3226. BIBP3226 alone did not alter CRF release from hypothalamic explants. Orexin-A had no effect in vitro on the release of other neuropeptides, AVP and noradrenaline, involved in the central regulation of the HPA axis. These results suggest that orexin-A is involved in activation of the HPA axis, and that these effects could be mediated via the release of NPY. PMID- 11412345 TI - Evaluation of a reversed passive latex agglutination test for the detection of Verocytotoxin (VT) expressed by strains of VT-producing Escherichia coli. AB - AIMS: To compare an experimental Reversed-Passive Latex Agglutination (RPLA) with Vero cells for the detection of Verocytotoxin expressed by VT-producing strains of Escherichia coli (VTEC). METHODS AND RESULTS: The RPLA was used alongside a Vero cell tissue culture assay for the detection of VT in bacterial culture supernatant fluids and patients' faecal extracts. CONCLUSION: The RPLA was comparable with the Vero cell assay, although slightly less sensitive. SIGNIFICANCE AND IMPACT OF THE STUDY: The RPLA test proved to be a simple, rapid and convenient method of detecting VT in bacterial culture supernatant fluids and in the faeces of patients infected with VTEC. PMID- 11412346 TI - Transmission of Yersinia enterocolitica 4/O:3 to pets via contaminated pork. AB - AIMS: This study was conducted to investigate sources of Yersinia enterocolitica 4/O:3 infections in dogs and cats. METHODS AND RESULTS: Transmission of Y. enterocolitica 4/O:3 to pets via contaminated pork was studied using PFGE with NotI, ApaI and XhoI enzymes. A total of 132 isolates, of which 16 were from cat and dog faeces and 116 from raw pork samples, were recovered in Finland during 1998-99. Cat 1, whose diet consisted mostly of raw pig hearts and kidneys, excreted Y. enterocolitica 4/O:3 of genotype G4. This predominant genotype was also found in isolates recovered from the pig heart, liver, kidney, tongue and ear, and minced pork samples. Dog 2, which was fed raw minced pork, excreted Y. enterocolitica of genotype G13. This genotype was also identified in isolates recovered from the pig heart, kidney and tongue, and minced pork samples. CONCLUSION: These results show that raw pork can be an important source of Yersinia enterocolitica 4/O:3 infections in dogs and cats. SIGNIFICANCE AND IMPACT OF THE STUDY: Raw pork should not be given to pets. PMID- 11412347 TI - Tolerance of Bradyrhizobium sp. (Lupini) strains to salinity, pH, CaCO3 and antibiotics. AB - AIMS: Ten rhizobial isolates obtained from different locations in Egypt were examined for their ability to survive under stress conditions and their growth response to increasing levels of NaCl (1-8% w/v), pH (4-10), CaCO3 (1-10% w/v) and 12 antibiotics. METHODS AND RESULTS: All the rhizobial isolates tolerated a NaCl concentration up to 5% and were divided into two groups with respect to NaCl tolerance. The rhizobial isolates from group two showed significantly (P < 0.05) better survival under high NaCl concentration. All the tested isolates survived acidic (pH 4-5) and alkaline conditions (pH 9-10) and CaCO3 (up to 10% w/v) in liqued YEM medium. CONCLUSION: Antibiotic resistance patterns did not correlate to NaCl, pH or CaCO3 tolerance. Variations among different strains showed that there is potential to improve strain performance under stress conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that selection of adapted strains under stress conditions is possible and can be used as inoculants for successful lupin growth. PMID- 11412348 TI - Use of temporal temperature gradient gel electrophoresis to identify flaA and fim3 sequence types in Bordetella bronchiseptica. AB - AIMS: The aim of this study was to develop an approach to detect variation in the flaA and fim3 genes amongst animal isolates of Bordetella bronchiseptica using temporal temperature gradient gel electrophoresis (TTGE). METHODS AND RESULTS: Amplicons representing three flaA and two fim3 sequence types were subjected to TTGE analysis. It was possible to distinguish clearly between each of the sequence types using TTGE. CONCLUSION: This suggests that TTGE could be a useful tool for studying the epidemiology of B. bronchiseptica. SIGNIFICANCE AND IMPACT OF THE STUDY: PCR amplification coupled to TTGE offers a general method for the rapid screening of large numbers of microbial strains for variations in gene sequences. PMID- 11412349 TI - In situ reverse transcription to detect the cbbL gene and visualize RuBisCO in chemoautotrophic nitrifying bacteria. AB - AIMS: In situ methodologies targeting the cbbL gene were used to visualize cells of nitrifying bacteria. Both procaryotic in situ PCR (IS-PCR) and in situ reverse transcription (ISRT) protocols were employed to determine gene presence and expression, respectively. METHODS AND RESULTS: Aged-oligotropic seawater samples were inoculated with microbial assemblages containing a mixture of actively growing nitrifying bacteria, starved nitrifying bacteria, and heterotrophic bacteria without cbbL. After the molecular manipulations, we found that while all the nitrifiers (healthy or starved) with the cbbL gene were detected by IS-PCR, only the actively growing autotrophic nitrifiers with detectable levels of carbon fixation and nitrification activity were detected by ISRT analysis. CONCLUSION: These results show how IS-PCR and ISRT supplement each other, and their potential for the analysis of heterogeneous populations where an assortment of healthy and starved/dormant cells are expected. PMID- 11412350 TI - Identification of Streptococcus salivarius by PCR and DNA probe. AB - AIMS: To establish species-specific PCR and DNA probe methods for Streptococcus salivarius and to clarify the distribution of dextranase in oral isolates of Strep. salivarius. METHODS AND RESULTS: A pair of PCR primers and a DNA probe were designed based on the nucleotide sequence of the dextranase gene of Strep. salivarius JCM5707. Both the PCR primer and the DNA probe specifically detected Strep. salivarius but none of the other oral streptococci (23 strains of 13 species). The primer and the probe were capable of detecting 1 pg and 1 ng of the genomic DNA, respectively, purified from Strep. salivarius JCM5707. All oral isolates (130 strains from 12 subjects) of Strep. salivarius from human saliva were positive by both methods. CONCLUSION: The present PCR and DNA probe methods are highly specific to Strep. salivarius and are useful for the its detection and identification of this bacterium. The dextranase widely distributes among oral isolates of Strep. salivarius. SIGNIFICANCE AND IMPACT OF THE STUDY: The DNA sequence of a dextranase gene present in the genome of Strep. salivarius is useful as the target DNA of the species-specific PCR and DNA probe. PMID- 11412351 TI - (13)C incorporation into DNA as a means of identifying the active components of ammonia-oxidizer populations. AB - AIMS: To identify active CO2-assimilating species of ammonia-oxidizing bacteria in fresh water sediment. METHODS AND RESULTS: Enrichment cultures were incubated in the presence of 13C labelled CO2, and 13C-DNA successfully resolved from 12C DNA by caesium chloride density gradient ultracentrifugation of DNA extracts. Ammonia-oxidizer DNA recovered from these gradients was amplified and characterised by Temporal Temperature Gradient Gel Electrophoresis (TTGE), with confirmatory sequence analysis to identify the metabolically active components of the population. CONCLUSION: The 12C-DNA fraction was dominated by nitrosospiras, in contrast to the 13C-DNA fraction which was largely nitrosomonad DNA, in support of the hypothesis that nitrosomonads out-compete nitrosospiras in laboratory culture. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of stable isotype incorporation into ammonia-oxidizer DNA could therefore circumvent the problems associated with RNA detection to identify metabolically active species in situ. PMID- 11412352 TI - Enhancement of cephalosporin C production by cultivation of Cephalosporium acremonium M25 using a mixture of inocula. AB - AIMS: To enhance the productivity of Cephalosporin C (CPC) by cultivation of Cephalosporium acremonium M25 using a mixture of inocula. METHODS AND RESULTS: Inoculum age was classified into three stages (early, intermediate and late) by image analysis. A mixture of inocula, according to the inoculum ages, was used for efficient production of CPC in the main culture. The most effective mixing ratio of inocula for CPC production in shake flasks was a 3 : 7 volume ratio of early- and late-stage inocula. This was also the case in a 1.5 l stirred-tank reactor. CPC productivity was enhanced by about 32% and 34% when using an inoculum mixture in the shake flask and 1.5 l stirred-tank reactor, respectively. CONCLUSION: The morphological characteristics of C. acremonium M25 in the seed culture were quite different according to inoculum age. The compromise of different ages of inoculum showed better production of CPC. SIGNIFICANCE AND IMPACT OF THE STUDY: The productivity of CPC was enhanced considerably when using mixed inocula. The results of this study can be applied to fungal cultures for efficient production of various metabolites. PMID- 11412353 TI - Laccase production by Phanerochaete chrysosporium--an artefact caused by Mn(III)? AB - AIMS: The possibility of laccase production by Phanerochaete chrysosporium was studied. METHODS AND RESULTS: A relatively high initial Mn(II) concentration (1-4 mM) in the growth medium leads to the development of reddish-brown coloration and intensive oxidation of 2.2'-azino-bis(3-etilbenz-tiazolin-6-sulfonate) (ABTS). The peak of ABTS oxidation was obtained approximately 1 day after the peak of MnP activity. CONCLUSION: ABTS oxidation was not caused by manganese peroxidase (MnP) nor by laccase but was the consequence of the action of Mn(III) which was stabilised in the growth medium. Decomposition of the complex took place after the biomass was removed from the growth medium and especially after the aeration of the culture was interrupted. SIGNIFICANCE AND IMPACT OF THE STUDY: Mn(III) seems to be the cause of false positive laccase reactions. More reliable data on MnP activity can be obtained if the complex is decomposed by the fungus before MnP activity is measured in the medium. PMID- 11412354 TI - Inducible and constitutive expression using new plasmid and integrative expression vectors for Thermus sp. AB - AIMS: To develop molecular tools and examine inducible and constitutive gene expression in Thermus thermophilus. METHODS AND RESULTS: Two plasmid promoter probe vectors and an integrative promoter probe vector were constructed using a promoterless thermostable kanamycin nucleotidyltransferase (KmR) cassette. Three expression vectors were constructed based on a constitutive promoter J17, that functions in both Thermus and Escherichia coli. An inducible expression vector was constructed using the heat-shock inducible promoter (70 to 85 degrees C) from the dnaK gene of T. flavus, and the malate dehydrogenase gene (mdh) from T. flavus was cloned and expressed in both E. coli and T. thermophilus HB27. CONCLUSION: This report describes the construction and use of improved promoter probe and expression vectors for use in Thermus species. The mdh gene can be used as a high temperature (85 degrees C) reporter gene for Thermus sp. The dnaK promoter is thermo-inducible. SIGNIFICANCE AND IMPACT OF THE STUDY: The expression vectors and molecular tools described here are significant improvements over previously reported vectors for Thermus sp. The mdh gene and the thermo-inducible dnaK promoter will facilitate high temperature studies employing Thermus species. PMID- 11412355 TI - Characterization and control of thread mould in cheese. AB - AIMS: The origin of a mould responsible for the contamination of an Argentinian cheese factory was identified and several antifungal treatments were assessed. METHODS AND RESULTS: Moulds were isolated and identified from vacuum-packed hard cheeses, from the environment and from the surfaces of the factory. A suspension conidia test containing different fungicides was performed; another assay involved the fumigation with p-OH fenilsalicidamide. Only Phoma glomerata was found in all of the mouldy cheeses, and was also obtained from different environments and machine surfaces. The most effective treatments against P. glomerata isolates were 0.5% (w/v) natamycin and 2% (v/v) parabens. Fumigation with p-OH fenilsalicidamide showed no satisfactory results. CONCLUSION: P. glomerata is an important thread mould-contaminating agent in vacuum-packed hard cheeses. SIGNIFICANCE AND IMPACT OF THE STUDY: Taking into account the survival of the conidia of the P. glomerata isolates to different antifungal treatments, the sources of contamination need to be controlled by designing a good factory layout. PMID- 11412356 TI - recA genotyping of Salmonella enteritidis phage type 4 isolates by restriction fragment length polymorphism analysis. AB - AIMS: To subtype Salmonella enteritidis phage type 4 isolates by using recA genotyping. METHODS AND RESULTS: Random amplified polymorphic DNA analysis using a primer ERIC2 of 76 isolates of Salmonella enteritidis phage type 4 obtained in Northern Ireland in 1998 and in 1999 demonstrated the presence of five genotypes. Restriction fragment length polymorphism analysis, using a degenerate primer pair designed to amplify a segment (about 640 bp in length) of the recA gene from several members of the Enterobacteriaceae with restriction enzymes, HhaI and Sau3AI, showed that the resulting fragments could differentiate the isolates into three groups, respectively. CONCLUSION: recA gene amplification and HhaI and Sau3AI restriction digestion was demonstrated to increase the differentiating power between isolates of Salmonella enteritidis phage type 4 by combining the patterns of the random amplified polymorphic DNA analysis procedure using a primer ERIC2. SIGNIFICANCE AND IMPACT OF THE STUDY: A novel restriction fragment length polymorphism assay for isolates of Salmonella enteritidis phage type 4, based on the amplification of the recA gene was attained and its comparison and its combination with random amplified polymorphic DNA analysis was provided. PMID- 11412357 TI - Influence of strain and trisodium phosphate concentration on growth parameters of Listeria monocytogenes in vitro. AB - AIMS: The present study was conducted to determine the influence of strain and trisodium phosphate (TSP) concentration in the growth of Listeria monocytogenes in vitro. METHODS AND RESULTS: Three strains (ATCC 11916, 64d, isolated from chicken meat, and M2-5b, a clinical animal isolate) were inoculated in broth with 0, 0.5, 1 and 1.5% (w/v) of TSP. The shortest lag phase and highest maximum rate of growth (mu) were obtained in the presence of 0.5% TSP. In contrast, the highest lag phase and lowest mu were obtained with 1.5% TSP. For each TSP concentration, significant differences (P < 0.05) in lag phase and mu of the three L. monocytogenes strains were observed. CONCLUSION: The behaviour of L. monocytogenes is significantly influenced by both the origin of the strain and the salt concentration. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results indicate the importance of choosing an adequate TSP concentration for the decontamination of foods, because low concentrations could favour the growth of L. monocytogenes. PMID- 11412358 TI - Exopolysaccharide-producing strains of thermophilic lactic acid bacteria cluster into groups according to their EPS structure. AB - AIMS: To compare galactose-negative strains of Streptococcus thermophilus and Lactobacillus delbrueckii subspecies bulgaricus isolated from fermented milk products and known to produce exopolysaccharides (EPSs). METHODS AND RESULTS: The structures of the EPSs were determined using nuclear magnetic resonance (NMR) and their genetic relationships determined using restriction endonuclease analysis (REA) and random amplification of polymorphic DNA (RAPD). Similar groupings were apparent by REA and RAPD, and each group produced an EPS with a particular subunit structure. CONCLUSION: Although none of the strains assimilated galactose, all inserted a high proportion of galactose into their EPS when grown in skimmed milk, and fell into three distinct groups. SIGNIFICANCE AND IMPACT OF THE STUDY: This information should help in an understanding of genetic exchanges in lactic acid bacteria. PMID- 11412359 TI - Origin of honeybees (Apis mellifera L.) from the Yucatan peninsula inferred from mitochondrial DNA analysis. AB - Honeybees (Apis mellifera L.) sampled at sites in Europe, Africa and South America were analysed using a mitochondrial DNA restriction fragment length polymorphism (RFLP) marker. These samples were used to provide baseline information for a detailed analysis of the process of Africanization of bees from the neotropical Yucatan peninsula of Mexico. Radical changes in mitochondrial haplotype (mitotype) frequencies were found to have occurred in the 13-year period studied. Prior to the arrival of Africanized bees (1986) the original inhabitants of the Yucatan peninsula appear to have been essentially of southeastern European origin with a smaller proportion having northwestern European ancestry. Three years after the migration of Africanized bees into the area (1989), only very low levels of maternal gene flow from Africanized populations into the resident European populations had occurred. By 1998, however, there was a sizeable increase in the proportion of African mitotypes in domestic populations (61%) with feral populations having 87% of mitotypes classified as African derived. The results suggest that the early stages of Africanization did not involve a rapid replacement of European with African mitotypes and that earlier studies probably overestimated the prevalence of African mitotypes. PMID- 11412360 TI - Bottlenecks, drift and differentiation: the population structure and demographic history of sika deer (Cervus nippon) in the Japanese archipelago. AB - We assessed genetic differentiation and diversity in 14 populations of sika deer (Cervus nippon) from Japan and four populations of sika deer introduced to the UK, using nine microsatellite loci. We observed extreme levels of differentiation and significant differences in diversity between populations. Our results do not support morphological subspecies designations, but are consistent with previous mitochondrial DNA analyses which suggest the existence of two genetically distinct lineages of sika deer in Japan. The source of sika introduced to the UK was identified as Kyushu. The underlying structure of Japanese populations probably derives from drift in separate glacial refugia and male dispersal limited by distance. This structure has been perturbed by bottlenecks and habitat fragmentation, resulting from human activity from the mid-nineteenth century. Most current genetic differentiation and differences in diversity among populations probably result from recent drift. Coalescent model analysis suggests sika on each of the main Japanese islands have experienced different recent population histories. Hokkaido, which has large areas of continuous habitat, has maintained high levels of gene flow. In Honshu the population is highly fragmented and is likely to have been evolving by drift alone. In Kyushu there has been a balance between gene flow and drift but all the populations have experienced high levels of drift. Habitat fragment size was not significantly associated with genetic diversity in populations but there was a significant correlation between habitat fragment size and effective population size. PMID- 11412361 TI - Spatial effects and rare outcrossing events in Medicago truncatula (Fabaceae). AB - In order to elucidate the mechanisms underlying the large amount of RAPD polymorphism found in 1990 in a population of the selfing annual Medicago truncatula GAERTN. (Fabaceae), we have analysed most of the individuals (n = 363) from the same population 6 years later using microsatellite loci. We confirm the result of the earlier study, namely that this population is very polymorphic and highly subdivided, with approximately 37% of the variance distributed among subpopulations, only 50 m apart one from another. We use standard F-statistics analyses, linkage disequilibria, minimum spanning network, multilocus assignment tests and spatial autocorrelation analyses to test the hypotheses that spatial structure and outcrossing events are involved in maintaining the large amount of genetic diversity at the level of each subpopulation. Interestingly, fine-scale spatial structure could be observed in only one subpopulation suggesting that other mechanisms are acting elsewhere. To the best of our knowledge, this is the first study of fine spatial genetic structure in a predominantly selfing species. PMID- 11412362 TI - The extent of clonality and genetic diversity in lingonberry (Vaccinium vitis idaea L.) revealed by RAPDs and leaf-shape analysis. AB - Numerous plant species reproduce mainly by clonal growth, implying that levels of genetic variation may be comparatively low. In this study we describe the genetic and genotypic diversity within and between four Swedish populations of the clonal shrub lingonberry, Vaccinium vitis-idaea L. Two approaches were used to assess the amount and partitioning of variation: automated image analysis of leaf shape and random amplified polymorphic DNA (RAPD) analysis. Morphometric analyses, using moment invariants and elliptic Fourier coefficients, revealed that most of the variation could be attributed to within-population variation. With the use of 43 polymorphic RAPD markers, we were able to identify 29 different genotypes (i.e. putative clones) among 129 plants from two populations. The genotypic diversity (D: mean 0.84) and evenness (E: mean 0.81) were higher than the average for clonal plant species. Within-population gene diversity was similar to values reported in nonclonal plants, suggesting that sexual reproduction has played a significant role in these populations despite low levels of seedling recruitment in present-day populations. An analysis of molecular variance revealed that most of the variation (89.2%) resided within populations. Comparisons between the different suites of characters indicated a congruent pattern of partitions of diversity, particularly when comparing moment invariants and RAPDs. When comparing the ability of the two descriptor suites to assign the plants to the RAPD-defined clones, elliptic Fourier coefficients yielded the best result; a classification test correctly reassigned 96.9% and 98.1% of the plant material in the two respective populations. PMID- 11412363 TI - Using microsatellite and MHC variation to identify species, ESUs, and MUs in the endangered Sonoran topminnow. AB - Highly variable loci can provide insight into the recognition of species, evolutionarily significant units (ESUs) and management units (MUs). In general, the ESU and MU categories are thought to be reflective of adaptive differences between them. Here we examine this premise by presenting a comprehensive examination of genetic variation for both microsatellite loci and a major histocompatibility complex (MHC) locus, thought to be of adaptive significance, in the endangered Sonoran topminnow. The extent of variation for the microsatellite loci and the MHC gene within the 13 populations of the Gila topminnow is highly correlated, suggesting that nonselective factors have played an important role in influencing variation within and between populations for the MHC locus. Therefore, using all of these loci, we found that the eight natural populations of the Gila topminnow fell into two different ESUs, one of which had four different MUs. The source of the Boyce Thompson sample, a population that was used extensively for restocking, appeared to be Monkey Spring. The source of the Watson Wash population also appeared to be Monkey Spring (or Boyce Thompson). The newly colonized Santa Cruz River population, which had the most genetic variation of any Gila topminnow population, appeared to descend primarily from Sonoita Creek populations. The Yaqui topminnow, presently considered another subspecies of the Sonoran topminnow, was very distinct for both microsatellite (only two of 25 alleles found in the Yaqui were in any of the Gila topminnow samples) and MHC alleles (nonoverlapping sets of alleles for the two groups). As a result, it appeared that the taxonomic status of the two subspecies should be re-evaluated and that full species status for Gila and Yaqui topminnows was appropriate. There was evidence for the importance of long-term selection at the MHC locus in the higher rate of nonsynonymous than synonymous substitution. In addition, there appeared to have been a duplication of the MHC locus that was present in most of the fish in six of the natural populations of the Gila topminnow. PMID- 11412364 TI - Spatial patterns of genetic diversity in Posidonia oceanica, an endemic Mediterranean seagrass. AB - Posidonia oceanica is an endemic seagrass species in the Mediterranean Sea. In order to assess levels of genetic structure in this species, the microsatellite polymorphism was analysed from meadows collected in several localities, along the coasts of the Tyrrhenian Sea (Mediterranean Sea). The existence of single population units and the recruitment of seedlings collected in some localities were investigated. Moreover, genetic structure at different spatial scales and biogeographic relationships among populations were also assessed. Our analysis showed the existence of clear patterns of genetic structure in P. oceanica in the area considered in the analysis. P. oceanica, in fact, is present in separate meadows that represent discrete populations, characterized by low genetic diversity. Comparable levels of genetic variability between mature meadows and seedlings were found. Patterns of genetic relatedness among populations seem to be in accord with direction of dominant current flux in the whole area, separating South Tyrrhenian from North Tyrrhenian populations. Moderate levels of gene flow between populations and genetic substructure within populations, together with the finding of the limited role of sexual reproduction in increasing genetic variability, should be a cause for concern for the persistence of this essential resource in the Mediterranean basin. PMID- 11412365 TI - Hierarchical genetic structure of the introduced wasp Vespula germanica in Australia. AB - The wasp Vespula germanica is a highly successful invasive pest. This study examined the population genetic structure of V. germanica in its introduced range in Australia. We sampled 1320 workers and 376 males from 141 nests obtained from three widely separated geographical areas on the Australian mainland and one on the island of Tasmania. The genotypes of all wasps were assayed at three polymorphic DNA microsatellite markers. Our analyses uncovered significant allelic differentiation among all four V. germanica populations. Pairwise estimates of genetic divergence between populations agreed with the results of a model-based clustering algorithm which indicated that the Tasmanian population was particularly distinct from the other populations. Within-population analyses revealed that genetic similarity declined with spatial distance, indicating that wasps from nests separated by more than approximately 25 km belonged to separate mating pools. We suggest that the observed genetic patterns resulted from frequent bottlenecks experienced by the V. germanica populations during their colonization of Australia. PMID- 11412366 TI - Genetic structure of populations of the human hookworm, Necator americanus, in China. AB - Twenty-one to 58 individual Necator americanus were sampled from each of four villages in south-western China. Each nematode was sequenced for 588 bp of the mitochondrial cytochrome oxidase I gene. Allelic and nucleotide diversity varied two-fold among villages. Overall FST among populations was approximately 0.28, but this large value resulted from one low-diversity population that had a large genetic distance to the other three populations (F(ST) = 0.10 without that population). There was no correlation between geographical and genetic distance among sites. Thus, the genetic structure of this species in China may be characterized by variable effective sizes and uneven movement among sites. We discuss the implications of this genetic structure for vaccine development and the spread of drug resistance in human hookworms, and compare the genetic structure of hookworms with that of other nematodes. PMID- 11412367 TI - Population subdivision in marine environments: the contributions of biogeography, geographical distance and discontinuous habitat to genetic differentiation in a blennioid fish, Axoclinus nigricaudus. AB - The relative importance of factors that may promote genetic differentiation in marine organisms is largely unknown. Here, contributions to population structure from a biogeographic boundary, geographical distance and the distribution of suitable habitat were investigated in Axoclinus nigricaudus, a small subtidal rock-reef fish, throughout its range in the Gulf of California. A 408-bp fragment of the mitochondrial control region was sequenced from 105 individuals. Variation was significantly partitioned between 28 of 36 possible combinations of population pairs. Phylogenetic analyses, hierarchical analyses of variance and a modified Mantel test substantiated a major break between two putative biogeographic regions. This genetic discontinuity coincides with an abrupt change in ecological characteristics, including temperature and salinity, but does not coincide with known oceanographic circulation patterns or any known historic barriers. There was an overall relationship of increasing genetic distance with increasing geographical distance between population pairs, in a manner consistent with isolation-by-distance. A significant habitat-by-geographical-distance interaction term indicated that, for a given geographical distance, populations separated by discontinuous habitat (sand) are more distinct genetically than are populations separated by continuous habitat (rock). In addition, populations separated by deep open waters were more genetically distinct than populations separated by continuous habitat (rock). These results indicate that levels of genetic differentiation among populations of A. nigricaudus cannot be explained by a single factor, but are due to the combined influences of biogeography, geographical distance and availability of suitable habitat. PMID- 11412368 TI - Allozymic evidence of parapatric differentiation of brown trout (Salmo trutta L.) within an Atlantic river basin of the Iberian Peninsula. AB - The genetic variation of brown trout from Duero, one of the main Atlantic Iberian river basins, was assessed at 34 enzymatic loci in 62 native populations. A strong intrabasin differentiation was detected (G(ST) = 0.46; range D: 0-0.066), mainly attributable to the existence of two divergent groups of populations within Duero: southern and northern groups. This divergence was mainly a consequence of the unequal distribution of *75 and *100 alleles at sMDH-B1,2* isoloci, which were correlated with substantial differences in genetic diversity among regions. The Lower Course region (nearly fixed for the *100 allele) and Pisuerga River (nearly fixed for the *75 allele) showed lower heterozygosities (H approximately 0.8%) in contrast with adjacent areas, which evidenced intermediate frequencies for both alleles and higher heterozygosities (H: 2.2-3.1%). Vicariance appeared as the more probable explanation for the significant positive correlation detected between genetic and geographical distances in Duero Basin. Genetic relationships with adjacent Iberian drainages indicate a close similarity between the southern group and Cantabric trout, whereas the northern group constitutes an ancient form from this basin. This study confirmed complex genetic relationships in brown trout from northwest Iberia, reasserting the existence of clines at several loci and for genetic diversity. The interaction between Cantabric and Duero trout, as well as the location of the limit of the anadromous form around the 42 degrees N parallel, are both required to understand the genetic characteristics of brown trout from this area. PMID- 11412369 TI - Genetic and ecological divergence of a monophyletic cichlid species pair under fully sympatric conditions in Lake Ejagham, Cameroon. AB - Although there is mounting evidence that speciation can occur under sympatric conditions, unambiguous examples from nature are rare and it is almost always possible to propose alternative allopatric or parapatric scenarios. To identify an unequivocal case of sympatric speciation it is, therefore, necessary to analyse natural settings where recent monophyletic species flocks have evolved within a small and confined spatial range. We have studied such a case with a cichlid species flock that comprises five Tilapia forms endemic to a tiny lake (Lake Ejagham with a surface area of approximately 0.49 km2) in Western Cameroon. Analysis of mitochondrial D-Loop sequences shows that the flock is very young (approximately 10(4) years) and has originated from an adjacent riverine founder population. We have focused our study on a particular pair of forms within the lake that currently appears to be in the process of speciation. This pair is characterized by an unique breeding colouration and specific morphological aspects, which can serve as synapomorphic characters to prove monophyly. It has differentiated into a large inshore and a small pelagic form, apparently as a response to differential utilization of food resources. Still, breeding and brood care occurs in overlapping areas, both in time and space. Analysis of nuclear gene flow on the basis of microsatellite polymorphisms shows a highly restricted gene flow between the forms, suggesting reproductive isolation between them. This reproductive isolation is apparently achieved by size assortative mating, although occasional mixed pairs can be observed. Our findings are congruent with recent theoretical models for sympatric speciation, which show that differential ecological adaptations in combination with assortative mating could easily lead to speciation in sympatry. PMID- 11412370 TI - Chloroplast microsatellites and mitochondrial nad1 intron 2 sequences indicate congruent phylogenetic relationships among Swiss stone pine (Pinus cembra), Siberian stone pine (Pinus sibirica), and Siberian dwarf pine (Pinus pumila). AB - We studied the phylogenetic relationships among the three stone pine species, Pinus cembra, P. sibirica, and P. pumila, using chloroplast microsatellites and mitochondrial nad1 intron 2 sequences. The three chloroplast microsatellite loci combined into a total of 18 haplotypes. Fourteen haplotypes were detected in 15 populations of P. cembra and one population of P. sibirica, five of which were shared between the two species, and the two populations of P. pumila comprised four species-specific haplotypes. Mitochondrial intron sequences confirmed this grouping of species. Sequences of P. cembra and P. sibirica were identical, but P. pumila differed by several nucleotide substitutions and insertions/deletions. A repeat region found in the former two species showed no intraspecific variation. These results indicate a relatively recent evolutionary separation of P. cembra and P. sibirica, despite their currently disjunct distributions. The species-specific chloroplast and mitochondrial markers of P. sibirica and P. pumila should help to trace the hybridization in their overlapping distribution area and to identify fossil remains with respect to the still unresolved postglacial re-colonization history of these two species. PMID- 11412371 TI - Phylogeography and conservation genetics of the Columbia spotted frog (Rana luteiventris; Amphibia, Ranidae). AB - The Columbia spotted frog (Rana luteiventris) has a widespread distribution in western Canada and the western US, although the southern reach of its range is highly fragmented into several isolated populations. Threats from various factors have raised concerns regarding the long-term survival of many small, isolated populations. Here, we report a study designed to determine the phylogeographic and conservation genetic parameters of R. luteiventris in the western US. Mitochondrial DNA (mtDNA) sequences were examined for phylogeographic structuring using phylogenetic reconstruction methods, coupled with networking and nested clade analyses. These methods permitted a distinction to be made between historic and demographic forces acting to generate geographical patterning of genetic variation. Phylogenetic analysis revealed four geographically correlated monophyletic clades. Three of these clades correspond to well-defined, nonoverlapping geographical locations in the fragmented portion of the range. The other is comprised of all samples collected from the contiguous range and includes one isolate from northern Wyoming. Networking and nested clade analyses confirmed these results and revealed that historical processes, such as range expansion and vicariance, rather than recurrent gene flow are likely responsible for observed patterns of genetic variation. A measure of genetic variation (theta = 4N(e)mu) revealed that R. luteiventris populations in Utah have a relatively low amount of genetic variation compared with populations in the continuous portion of the range. PMID- 11412372 TI - Shared parentage and incest avoidance in the cooperatively breeding acorn woodpecker. AB - Social groups of acorn woodpeckers (Melanerpes formicivorus) range in size from unaided pairs to 15 adults. Behavioural indicators of mate guarding, assumed incest avoidance and observations of egg-laying indicate that social organization ranges from monogamous pairs to groups with up to seven male and three female putative cobreeders plus up to 10 nonbreeding helpers. In addition, groups occasionally lack a putative breeder throughout the breeding season. Here we report results from multilocus DNA fingerprinting of 372 nestlings from 123 nests in groups with putative cobreeders of one or both sexes. No extra-group fertilizations were found. Putative cobreeding males within social groups shared paternity. However, the most reproductively successful male was, on average, almost three times as successful as the next most successful and additional males only occasionally sired offspring. In contrast, cobreeding females shared parentage equally. Helpers never bred incestuously when their opposite-sex parent (or another relative, such as their uncle) held breeding status in the group. However, during breeding male vacancies, 14 nestlings were produced when helper males bred incestuously with their mother. Both male and female helpers usually became successful cobreeders with their same-sex parent following replacement of the opposite-sex breeder(s) by unrelated individuals. PMID- 11412373 TI - Sex-biased dispersal and a density-independent mating system in the Australian brushtail possum, as revealed by minisatellite DNA profiling. AB - Natal dispersal can have important effects on mammal population structure and dynamics following a local population crash. Such dispersal is of practical importance when applied to the control of pest species because dispersal may significantly, and undesirably, reduce the population recovery time following a control operation. The relative dispersal rate of the sexes is also critical because that too will affect the rate of population increase. Here, we describe a field experiment in which we reduce the density of two populations of the Australian brushtail possum, and use genetic similarity, as estimated by minisatellite DNA profiles, to investigate dispersal in the original (undisturbed) and recovering populations. Our results show that the genetic similarity within the undisturbed populations was lower between males than between females. Conversely, the genetic similarities between males and females in the two recovering populations were not significantly different, while relatedness among males was significantly higher in the recovering populations when compared with those in the pre-removal populations. These data indicate two important characteristics of dispersal in possums: (i) that dispersal in established populations is sex biased towards males; and (ii) that within the first 3 years following population control, 'the vacuum effect', whereby individuals from areas adjacent to a control area expand their home range and invade the depopulated area, is the most important factor in the re-colonization process for possums. We found no evidence that the mating system, which is polygynous, varied when the density was markedly reduced. These results indicate that drastic reductions in population density by conventional control will not affect the rate of spread of biological control agents that rely on sexual transmission for dissemination. PMID- 11412374 TI - A comparison of microsatellite-based pairwise relatedness estimators. AB - Studies of inbreeding depression or kin selection require knowledge of relatedness between individuals. If pedigree information is lacking, one has to rely on genotypic information to infer relatedness. In this study we investigated the performance (absolute and relative) of 10 marker-based relatedness estimators using allele frequencies at microsatellite loci obtained from natural populations of two bird species and one mammal species. Using Monte Carlo simulations we show that many factors affect the performance of estimators and that different sets of loci promote the use of different estimators: in general, there is no single best performing estimator. The use of locus-specific weights turns out to greatly improve the performance of estimators when marker loci are used that differ strongly in allele frequency distribution. Microsatellite-based estimates are expected to explain between 25 and 79% of variation in true relatedness depending on the microsatellite dataset and on the population composition (i.e. the frequency distribution of relationship in the population). We recommend performing Monte Carlo simulations to decide which estimator to use in studies of pairwise relatedness. PMID- 11412375 TI - Genetic structure of brown trout (Salmo trutta, L.) populations from south western France: data from mitochondrial control region variability. AB - The genetic relationships between several French Atlantic populations of brown trout were analysed using polymerase chain reaction-single-strand conformation polymorphism and sequencing of two parts of the mitochondrial control region. Ten mitochondrial haplotypes were identified, separated by a small number of mutations. The distribution of these haplotypes confirmed our hypothesis of the existence of two genetically well-differentiated groups of populations in this area, which has already been suggested using nuclear markers. Nuclear and mitochondrial markers, however, show different patterns of genetic differentiation: some populations which belong to one group according to allozyme results appear to be part of the other when using mitochondrial DNA. The origin of these different forms is discussed; they seem to belong to the Atlantic grouping. Nevertheless, the hypothesis of an ancestral and a modern form suggested for northern populations does not appear valid in this particular case. PMID- 11412376 TI - Spatial analysis of allozyme and microsatellite DNA polymorphisms in the land snail Helix aspersa (Gastropoda: Helicidae). AB - The genetic structure of the land snail Helix aspersa was investigated for 21 populations collected along a road located in the polders of the Bay of Mont Saint-Michel (Brittany, France), following a sampling scheme the area of which did not exceed 900 m in length. A total of 369 individuals were genotyped for five enzymatic markers and seven microsatellite loci. We used sequential hierarchical F-statistics at different spatial scales and spatial autocorrelation statistics to explore recent historical patterns involved in the observed genetic distribution. Whatever the statistics used, congruent levels of spatial genetic substructuring across loci were demonstrated, excepted for one allozyme locus. Overall spatial genetic arrangement matched in a substantial fashion theoretical predictions based on the limited dispersal power of land snails. Positive autocorrelation over short-distance classes may result from the development of genetically distinct patches of individuals organized in family-structured colonies. Therefore, spatial signatures of average I correlograms can be viewed as the expression of a stepping-stone model of population structure, sometimes involving external migrational events. Overall, the revealed pattern of population subdivision on a microgeographical scale was suggestive of a neighbourhood structure. Finally, microsatellite loci are especially suitable for the detection of small genetic clustering, and combining different classes of markers offers the potential to gain further insight into the description of spatial genetic variability over short temporal and geographical scales. PMID- 11412377 TI - Microsatellite diversity associated with ecological factors in Hordeum spontaneum populations in Israel. AB - Microsatellite diversity at 18 loci was analysed in 94 individual plants of 10 wild barley, Hordeum spontaneum (C. Koch) Thell., populations sampled from Israel across a southward transect of increasing aridity. Allelic distribution in populations was not distributed randomly. Estimates of mean gene diversity were highest in stressful arid-hot environments. Sixty-four per cent of the genetic variation was partitioned within populations and 36% between populations. Associations between ecogeographical variables and gene diversity, H(e), were established in nine microsatellite loci. By employing principle component analysis we reduced the number of ecogeographical variables to three principal components including water factors, temperature and geography. At three loci, stepwise multiple regression analysis explained significantly the gene diversity by a single principal component (water factors). Based on these observations it is suggested that simple sequence repeats are not necessarily biologically neutral. PMID- 11412378 TI - Phylogeography of Cape Verde Island skinks (Mabuya). AB - The Cape Verde Islands are of volcanic origin with most having appeared between the early Miocene and mid-Pleistocene. They contain six known species of Mabuya skinks. Phylogeographical relationships within and among the relatively widespread taxa M. stangeri, M. spinalis and M. delalandii were inferred, based on approximately 1 kbp of the cytochrome b gene (mitochondrial DNA). Reciprocal monophyly of M. spinalis and M. stangeri was established, which may have arisen from an early Pliocene/late Miocene cladogenetic event. Considerable between island sequence divergence was detected among M. spinalis, which appears to have colonized the older islands (Sal and Boavista) first. Much lower sequence divergence was found in M. delalandii, indicating a more recent range expansion. Here, evidence points to colonization of the younger islands of Brava and Fogo soon after appearance. There are similarities between some of the described patterns and those seen in lizards from the Canary Islands. PMID- 11412379 TI - Schistosoma japonicum egg granuloma formation in the interleukin-4 or interferon gamma deficient host. AB - The roles of interleukin (IL)-4 and interferon (IFN)-gamma in Schistosoma japonicum egg granuloma formation were investigated in cercariae-infected (infection model) or after implantation of laid parasite eggs (egg implantation model) in cytokine deficient mice. Two weeks after hepatic egg-implantation, a markedly decreased mononuclear cell infiltration and lack of multinuclear cell formation were characteristic features in IL-4 deficient mice. By 4 weeks (late stage), the cellular reactions around the eggs were negligible in the deficient mice. Compared to the controls, there was a drastic reduction in the production of the Th2 cytokines, IL-4, IL-5 and IL-13. MCP-1 levels were also significantly lowered. In mice experimentally infected with cercariae, granuloma cellularity in both the wild-type and IL-4 deficient mice at 45 days and 10 weeks postinfection was analogous to the egg implantation model at 2 and 4 weeks. Overall, the effects of IFN-gamma deficiency on granuloma induction differed markedly from the IL-4 results. Two weeks after egg implantation, IFN-gamma deficient mice showed suppressed neutrophil response and hepatic necrosis with confluent mononuclear cell infiltration along the outer layer of granulomas. By 4 weeks, there was a decrease in cell infiltration, fibrosis and MCP-1 production while IL-10 production increased. While these early characteristic features for IFN-gamma deficiency were common to both the egg implantation (at 2 and 4 weeks) and cercariae infection model (at 45 days), there was a surprising difference, i.e. marked fibrosis was found in the late stages (at 10 weeks postinfection) of cercariae-infected mice, but not in parasite egg implanted mice. Furthermore, while IL-13 levels were unchanged, both MCP-1 and IL-4 production were significantly lower at 10 weeks in comparison with wild-type. The present study clearly demonstrates the importance of both Th1 and Th2 cytokine responses in S. japonicum egg-induced granuloma formation. PMID- 11412380 TI - Effect of nitric oxide synthase inhibition on Schistosoma japonicum egg-induced granuloma formation in the mouse liver. AB - Nitric oxide (NO) plays diverse roles in a variety of pathological processes. We investigated the role of NO in Schistosoma japonicum egg-induced granuloma formation in a mouse hepatic model. Immunohistological analysis revealed that there is the most intense and extensive inducible nitric oxide (iNOS) expression 2 weeks after egg implantation, and thereafter it decreased considerably with time. Treatment with nitric oxide synthase inhibitors, NIL (L-N6- (iminoethyl) lysine) or N(omega)-nitro-L-arginine methyl ester (L-NAME), resulted in two different types of unusual granulomas at 2 weeks. One type showed suppressed fibrosis, while another showed foreign body-type multinuclear cell formation which frequently appeared particularly when 50 microg/ml NIL was given. At 3 weeks following treatment, fibrotic granulomas with scanty peripheral cellularity was obvious. However, there were no apparent changes after this period (at 4 weeks). Cytokine analysis in NIL-treated mice showed a significant increase of IL 4 and IL-13 production at 2 weeks. These findings indicated that nitric oxide contributes to granuloma development during the early stages, probably through the regulation of Th2 cytokine production. PMID- 11412381 TI - Interleukin-10 does not contribute to the pathogenesis of a virulent strain of Toxoplasma gondii. AB - Interleukin (IL)-10 is an inhibitor of cell mediated immunity and an antagonist of the development of protective immune responses associated with resistance to T. gondii. These observations led to the hypothesis that the production of IL-10 could contribute to the ability of T. gondii to replicate and survive in an immune competent host. To determine whether the production of IL-10 affects the ability of the RH strain of T. gondii to cause a lethal infection in mice, we compared the immune response to RH in IL-10+/+ and IL-10-/- BALB/c mice. Both groups of mice produced comparable amounts of IL-12 and interferon (IFN)-gamma and had similar mortality curves and parasite burdens. The use of green fluorescent protein-labelled parasites allowed us to infect IL-10+/+ and IL-10-/- mice and use a fluorescence-activated cell sorter to distinguish infected and uninfected populations of macrophages and compare their expression of CD80, CD86 and major histocompatibility complex (MHC) class II. Although infected cells expressed higher overall levels of these molecules than uninfected cells, there were no differences between cells isolated from IL-10+/+ and IL-10-/- mice. Taken together, these results indicate that IL-10 is not required for the virulence of the RH strain of T. gondii, nor is it involved in the regulation of the CD80, CD86 and MHC class II molecules during RH-infection. PMID- 11412382 TI - Resistance to Encephalitozoon intestinalis is associated with interferon-gamma and interleukin-2 cytokines in infected mice. AB - The understanding of the immunopathology of infections caused by microsporidia has pinpointed the importance of T cell-mediated immunity. The immunopathology caused by the interesting protozoan parasite Encephalitozoon intestinalis, a microsporidium pathogenic in man, is not clearly understood. In this study, we demonstrate that a specific cellular immune response is implicated in the control of microsporidiosis infection in mice. Interferon (IFN)-gamma receptor knockout mice (IFN-gamma R(o/o)) developed a chronic infection with E. intestinalis, whereas a transient infection developed in wild-type mice. Encephalitozoon intestinalis proteins induced proliferation of murine spleen and mesenteric lymph node cells collected from infected mice. The host response to microsporidia infection was regulated by a specific pattern of cytokine protection. Spleen cells derived from resistant 129 Sv/Ev mice inoculated with E. intestinalis secreted significant levels of gamma-interferon and interleukin-2 but cells from highly susceptible IFN-gamma R knockout mice secreted high levels of interleukin 4 (mostly between 2 and 4 weeks post infection). This is the first report in which a specific cellular immune response against E. intestinalis infection is presented. PMID- 11412383 TI - Suppression of macrophage interleukin-12 and tumour necrosis factor-alpha production in mice infected with Toxocara canis. AB - Toxocara canis induces a predominantly Th2 type response with enhanced amounts of interleukin (IL)-4 and reduced amounts of interferon (IFN)-gamma in infected mice. In this study, we investigated the macrophage function of T. canis-infected mice from the standpoint of cytokine production. C3H/HeN mice were infected with T. canis by oral administration of embryonated eggs. Ten days after infection, macrophages were obtained from spleen and peritoneal cavity, were cultured with lipopolysaccharide, and cytokines in the culture supernatant were evaluated with enzyme-linked immunosorbent assay. Macrophages from T. canis-infected mice produced IL-1 and IL-6 equivalent to macrophages from normal mice. The production of IL-10 and tumour growth factor (TGF)-beta was enhanced, but IL-12 and tumour necrosis factor (TNF)-alpha production was diminished. The addition of IFN-gamma, anti-IL-10 antibody, anti-TGF-beta antibody or indomethacin did not restore the production of IL-12 and TNF-alpha by macrophages from T. canis-infected mice. Furthermore, the stimulation of normal macrophages with T. canis antigen in vitro induced IL-1alpha, IL-6, IL-10 and TGF-beta, but not IL-12 and TNF-alpha. These results indicate that cytokine producing pattern of macrophages is altered by T. canis-infection, and this altered macrophage function may play an important role in the modification of the immune response to T. canis. PMID- 11412384 TI - Characterization of two monoclonal antibodies raised in Btk(xid) mice that recognize phosphorylcholine-bearing antigens from Trichinella and other helminths. AB - This study investigated the binding properties of two monoclonal antibodies (mAbs US1 and US2) raised in (CBA/n x BALB/c)F1 (NBF1) Btk(xid) male mice. Both mAbs show unusual specificity for phosphorylcholine (PC)-containing TSL4 antigens of Trichinella. Specifically, and in contrast to mAbs raised in normal mice, US1 and US2 mAbs do not bind to artificial PC-protein conjugates and are not inhibited by either free PC or NPPC, although US2 was partially inhibited by NPPC at high concentration (10(-2) M). However, both mAbs completely abrogate the binding to Trichinella antigens of other anti-PC mAbs (e.g. BH8 and Mab-2). These results suggest that both US1 and US2 recognize complex PC-containing epitopes. The patterns of recognition of PC-bearing antigens from different helminths by US1, US2, Mab-2 and BH8 were broadly correlated with phylogenetic proximity. The closest similarities were observed between the members of the Trichinelloidea superfamily (Trichinella spiralis and Trichuris muris) and among the ascarids (Toxocara canis, Anisakis simplex, Hysterothylacium aduncum and Ascaris lumbricoides). However, US1 did not react with the filarial nematode Onchocerca volvulus and reacted only weakly with Onchocerca gibsoni, while US2 reacted only weakly with both species. Only BH8 recognized PC-bearing antigens from the trematode Fasciola hepatica and the cestode Bothriocephalus scorpii. These results suggest that PC is attached to identical or very similar structures on most different nematode species, although major differences exist with respect to helminth species from groups such as the trematodes and cestodes that are phylogenetically distant from the nematodes. PMID- 11412385 TI - [Methylation status of c-fms oncogene in hepatocellular carcinoma and its relation with clinical pathology]. AB - OBJECTIVE: To study the methylation status change of c-fms oncogene in hepatocellular carcinogenesis, and to clarify the abnormal highly expressing mechanism of c-fms/CSF-1R in hepatocellular carcinoma (HCC). METHODS: Genome DNA of 30 cases of HCC tissue and matching surrounding-cancer tissue were digested with restrictive endonucleases Hpa II/Msp I. Methylation status of c-fms oncogene was tested with Southern blot. Methylation status of c-fms oncogene in the HCC tissue and the matching surrounding-cancer tissue was compared. The relation between methylation status of c-fms oncogene and clinical pathology of HCC was determined. RESULTS: Methylation status of c-fms oncogene decreased in 36.7% (11/30) HCC tissue and 13.3% (4/30) matching surrounding-cancer tissue. The hypomethylation rate of c-fms oncogene in the HCC tissue was higher than that in the matching surrounding-cancer tissue. There was significant correlation between Edmondson scale of HCC and hypomethylation of c-fms oncogene. The hypomethylation rate of HCC in Edmondson III-IV was higher than that in Edmondson I-II. CONCLUSIONS: Hypomethylation of c-fms oncogene may be an important molecular mechanism leading to abnormally high expressing of CSF-1R, which contributes to occurrence and deterioration of HCC. PMID- 11412386 TI - [Evaluation of dynamic enhanced fast multiplanar spoiling gradient recalled(FMPSPGR) in the diagnosis of small hepatocellular carcinoma]. AB - OBJECTIVE: To analyze the features of small hepotocellular carcinoma (SHCC) by spin echo (SE) sequence and dynamic enhanced FMPSPGR, and to compare the sensitivity for detecting and the accuracy for characterization, further to discuss the value of FMPSPGR in the diagnosis of SHCC. METHODS: SE T(1)WI, T(2)WI and FMPSPGR dynamic imaging of the liver were performed for 58 patients with SHCC. The sensitivity for detecting and accuracy for characterization were statistically compared. RESULTS: Seventy-two lesions were found in 58 patients. In the detection of SHCC, the order of the sensitivity was dynamic enhanced FMPSPGR>SE T(2)WI>no-enhanced FMPSPGR>SE T(1)WI. Dynamic enhanced FMPSPGR was significantly higher than any of the others. There was significant difference between no-enhanced FMPSPGR and SE T(2)WI, but not significant difference between T(2)WI and no-enhanced FMPSPGR. In the characterization of SHCC, the accuracy of pre- and post-contrast FMPSPGR was significantly higher than that of SE (T(1)WI+T(2)WI) with remarkable statistical difference. CONCLUSIONS: FMPSPGR dynamic imaging is superior to SE in the detection and characterization of SHCC. The sensitivity for detecting and the accuracy for characterization will be improved when both FMPSPGR and SE are used. PMID- 11412387 TI - [Clinical significance of differential display cDNA fragment MRG98.2 expression in hepatocellular carcinoma]. AB - OBJECTIVE: To search for the gene related to hepatocellular carcinoma (HCC), and to study its potential significance in liver oncogenesis. METHODS: The difference in mRNA expression among HCC, nontumorous liver tissue, and normal liver tissue was investigated by differential display technique. Ten hepatic tissues of hepatoma versus surrounding noncancerous liver tissues were examined with MRG98.2 as a probe in Dot blot analysis. Expression of vascular endothelial growth factor (VEGF) mRNA in above-mentioned samples was examined with reverse transcription polymerease chain reaction (RT-PCR). RESULTS: Differential display cDNA fragment MRG98.2 was isolated from hepatoma samples. Dot blot analysis showed that MRG98.2 was expressed in 7 cases of HCC samples (7/10) but only in 2 of the surrounding noncancerous hepatic tissues (2/10). The expression of VEGF mRNA was upregulated in 6/7 of HCC samples expressed MRG98.2. CONCLUSIONS: MRG98.2 may be a gene related to HCC. Its expression in HCC correlates with VEGF mRNA. MRG98.2 expression is helpful in predicting a tendency toward invasion and metastasis of HCC or a poor prognosis. PMID- 11412388 TI - [Diagnostic value of five serum markers for liver fibrosis]. AB - OBJECTIVE: To research the diagnostic value of serum hyaluronic acid (HA), type III procollagen (PCIII), type IV collagen (CIV), laminin (LN), and transforming growth factor-beta(1) (TGF-beta(1)) for liver fibrosis in patients with chronic hepatitis. METHODS: Serum levels of HA, PCIII, CIV, LN and TGF-beta(1) in 116 patients with chronic hepatitis and cirrhosis were investigated and compared with hepatic histological findings of 87 patients. RESULTS: The correlation between serum HA and histologically assessed grade of inflammatory activity was weak (r=0.393, P<0.05). The correlation between serum HA, PCIII, LN, TGF-beta(1) and histologically assessed stage of liver fibrosis were all moderate (r=0.584, 0.454, 0.441 and 0.612, respectively, P<0.05), while that between serum CIV and histologically assessed stage of liver fibrosis was weak (r=0.319, P<0.05). As shown by the ROC curves in cases of chronic hepatitis, the ability to differentiate patients with cirrhosis from those without cirrhosis was greater for serum HA than that for serum PCIII, CIV, LN, and TGF-beta(1) (the areas under the curves=0.904 vs 0.784, 0.815, 0.805, 0.828, P<0.05). The ability of serum HA, LN and TGF-beta(1) to differentiate patients with extensive liver fibrosis from those with no or mild liver fibrosis exceeded that of serum PCIII and CIV (the areas under the curves=0.849, 0.819, 0.836 vs 0.702, 0.721, P<0.05). To discriminate the stage of liver fibrosis, serum HA and TGF-beta(1) were selected from the five markers by Bayes discriminate analysis. If S(1), S(2) and S(3) were not required to be discriminated, discrimination between the three stages showed significant difference (P<0.05). The predictive accurate percentage was 72.90%. CONCLUSIONS: The five markers all have the ability not only to judge liver cirrhosis, in which the ability of serum HA is the best, but also to differentiate chronic hepatitis with extensive liver fibrosis from that with no or mild liver fibrosis, in which the ability of serum HA, LN, TGF-beta(1) is stronger than the other two. It is useful for detection of serum HA and TGF beta(1) at the same time to discriminate the histologically assessed stage of fibrosis. Serum HA and TGF-beta(1) can be helpful in discriminating patients of chronic hepatitis with "no liver fibrosis", "liver fibrosis but no cirrhosis" and "liver cirrhosis", but cannot discriminate them accurately as the histologically assessed stage of fibrosis. They cannot displace liver biopsy for the judgement of liver fibrosis. PMID- 11412389 TI - [Study of the cytotoxity against human hepatocellular carcinoma cells induced by the MAGE-1 gene modified dendritic cells]. AB - OBJECTIVE: To genetically modify dendritic cells (DC) with a tumor associated antigen gene MAGE-1 and to observe in vitro the cytotoxic effect induced by this genetically modified DC against human hepatocellular carcinoma (HCC) cell line SMMC7721, thus giving a primary verification of the possibility of this genetically modified DC to induce specific antitumor immunity against HCC and serving as a new type of vaccine. METHODS: The MAGE-1 gene was inserted into the retrovirus vector LXSN to construct the recombinant retrovirus LMSN. The monocyte derived DCs were transfected by LMSN and control virus LXSN respectively as well as a third group was set up as non-transfected control. The MAGE-1 gene expression in LMSN transfected DC was identified by Western blot and the in vitro cytotoxities against SMMC7721 induced by three groups of DC were tested by MTT assay. RESULTS: A recombinant retrovirus LMSN containing the tumor rejection antigen gene MAGE-1 was successfully constructed and the MAGE-1 gene expression in LMSN transfected human monocyte-derived DC was induced. The LMSN transfected DC could induce very high lytic activity against SMMC7721 in vitro with the lytic activity of 78.9%+/-3.6%. Nevertheless, the LXSN transfected DC and non transfected DC only induced relatively lower lytic activity as 34.7%+/-4.3% and 3.9%+/-2.0%, respectively. The difference of the lytic activities between those three groups as statistically significant (P<0.01). CONCLUSIONS: The MAGE-1 gene transfected DC can induce higher in vitro cytotoxicity against SMMC7721, suggesting the possibility of this genetically modified DC to induce specific antitumor activity and to serve as a new type of vaccine for HCC. PMID- 11412390 TI - [Effects of interferon-alpha on recurrence and metastasis of hepatocellular carcinoma after curative resection in nude mice]. AB - OBJECTIVE: To study the effect of interferon-alpha (IFN-alpha) on the recurrence and metastasis of hepatocellular carcinoma (HCC) in nude mice, and to clarify if there is synergistic effect treated by combination of IFN-alpha and all-trans retinoic acid (ATRA). METHODS: The effect of IFN-alpha and/or ATRA on the proliferation of HCC cell lines was measured in vitro. The metastatic model of human HCC in nude mice LCI-D20 was used in present study. Curative resection was performed at 10th day after implantation in 44 nude mice. Drugs were given at the next day after resection. IFN-alpha was administered subcutaneously at doses of 3+/-10(5) U/day, 6+/-10(5) U/day, respectively. ATRA was administered p.o. at a dose of 20 mg/kg/day, and IFN-alpha 6+/-10(5)U/day combined with ATRA 20 mg/kg/day. The mice were sacrificed 35 days after treatment. The recurrent tumor size was measured and the presence of intrahepatic dissemination and lung metastases were recorded. RESULTS: The effect of IFN-alpha and/or ATRA on the proliferation of HCC cells SMMC7721, BEL-7402, BEL-7405, and MHCC97 was not obvious. The combination of IFN-alpha and ATRA had no synergistic effect in vitro. The lung metastatic rate, the liver recurrent rate, the size of main recurrent lesions, the number of intrahepatic disseminating nodules and the largest disseminating nodule of the controlled group was 100%(12/12), 100%(12/12), (1346.3+/-4.2 ) mm(3), 8.2+/-4.4, 864 mm(3), respectively; whereas it was 0, 87.5%(7/8), (8.7+/-2.9) mm(3), 2.3+/-0.6, and 7.8mm(3), respectively in the IFN-alpha 3+/-10(5) U/day treated group(P<0.05); 0, 12.5%(1/8), 0.5mm(3), 2, 0.5 mm(3) in the IFN-alpha 6+/-10(5)U/day treated group(P<0.05); 0, 12.5%(1/8), 1 mm(3), 2.5+/-0.7, 8 mm(3) in the IFN-alpha 3+/-10(5) U/day and ATRA treated group(P<0.05); 87.5%, 100%, (1472.6+/-5.6) mm(3), 7.3+/-3.8, 768.5 mm(3) in the ATRA treated group (P>0.05). CONCLUSIONS: IFN-alpha has an inhibitory effect on intrahepatic recurrence and lung metastasis of human HCC after curative resection in nude mice, and the effect is enhanced with increasing dose. IFN-alpha and ATRA have no synergistic effect according to in vivo and in vitro test. ATRA has no effect on recurrence and metastasis of HCC. PMID- 11412391 TI - [Reconstruction of hepatitis D virus ribozymes and their ability to trans-cleave hepatitis C virus RNA]. AB - OBJECTIVE: To study whether reconstructed hepatitis D virus (HDV) ribozymes have the ability to trans-cleave hepatitis C virus (HCV) RNA. METHODS: We reconstructed the stem IV and substrate-binding region of HDV genomic ribozymes, thus obtained three HDV ribozymes named RzC1, RzC2 and RzC3 expected to be able to trans-cleave HCV RNA at predicted sites. The substrate containing HCV RNA 5' noncoding region (5'-NCR) and 5'-fragment of C region (HCV RNA 5'-NCR-C) was synthesized by transcription in vitro, then radiolabelled at its 5'-end. Under certain pH and appropriate concentration of Mg(2+) with or without deionized formamide, the ribozymes and their substrate were mixed at mol ratio of 100 : 1 and reacted for two hours. Trans-cleaved products were shown by denatured polyacrylamide gel electrophoresis and autoradiography, and the percentage of trans-cleaved substrate was calculated as the activity indicator of these ribozymes. RESULTS: RzC1 and RzC2 were able to trans-cleave HCV RNA 5'-NCR-C site specifically, and their activity could be enhanced by certain concentration of deionized formamide. RzC3 could not trans-cleave the substrate. CONCLUSIONS: Optimized HDV genomic ribozymes can trans-cleave HCV RNA. PMID- 11412392 TI - [Determination of DNA binding domains in hepatitis C virus core protein]. AB - OBJECTIVE: To determine the DNA binding domain in hepatitis C virus core protein and otelucidate the significance of binding. METHODS: Segments of hepatitis C virus core protein were expressed in E.coli as fusion forms with glutathion S transferase (GST). The core proteins were immobilized in SDS-PAGE gel after removing SDS from the gel by washing. (32)P-ATP labeled oligonucleotides were electrophoresed through the gel in TAE buffer. The binding of DNA with core protein was detected by autoradiography. RESULTS: There were at least two DNA binding domains in the HCV core protein, the first locating at 10~16aa and the second at 46-70aa. The second region was divided into three adjacent parts, which could bind core protein independently. Core protein bound to single strand DNA as well as to double strand DNA without sequence specificity. CONCLUSIONS: DNA binding regions of HCV core protein locate at its N-terminus. The binding regions of HCV core protein overlap its nucleus transfer signals and they bind to target DNA unselectively, suggesting a possible mechanism for its multifunction. The result provides basic data for understanding the biological function of HCV core protein. PMID- 11412393 TI - [Cloning and sequence analysis of truncated S gene from circulation of patients with chronic hepatitis B virus infection]. AB - OBJECTIVE: To find different mutated status of HBV DNA in circulation from chronic HBV patients. METHODS: Specially designed primers and polymerase chain reaction method were applied to amplify the whole S gene of HBV from the serum of 2 patients. After being sequenced, 4 clones were compared with HBV adr subtype (China strain) to identify the mutant sites. RESULTS: Sequencing results implied that there was a truncated large/middle S gene in the serum of the patients. Besides that, HBsAg and HBV DNA polymerase defective clones were also detected. CONCLUSIONS: Truncated middle S gene is found in the circulation of patients with chronical HBV infection, suggestive of a poor prognosis. PMID- 11412394 TI - [Comparison of specific immune responses to duck hepatitis B virus in infected, immune, and uninfected ducks]. AB - OBJECTIVE: To explore the factor in determining whether hepadnavirus infection is cleared or becomes chronic. METHODS: Experimental groups were established by inoculation with duck hepatitis B virus (DHBV) at different age and schedule. The kinetics of virus replication and the humoral and cell mediated immune response (CMI) by ducks acutely and chronically infected with, or immune to DHBV infection was measured. RESULTS: Infection of the adolescent animals with DHBV led to a transient viremia. The levels of anti-DHBs and anti-DHBc were higher in acutely infected group than in chronically infected group (P<0.05), but lower than in immune group (P<0.01). CMI analysis showed the response to DHBsAg and DHBcAg by peripheral blood mononuclear cells from acutely infected ducks (10 day pi) was stronger than that from chronically infected ones (P<0.05); however, the level of the response reduced over a period of 5 weeks. There were no differences regarding CMI response in acutely infected or immune ducks. CONCLUSIONS: The immune response especially antigen -specific immune response is the key factor in determining the outcome of the infection. PMID- 11412395 TI - [Induction and kinetic characterization of nitric oxide synthase in hepatocytes]. AB - OBJECTIVE: To study the synergistic responses of nitric oxide synthase (NOS) induction in rat hepatocytes to LPS and various cytokines in vitro and the kinetic characteristics of iNOS. METHODS: The hepatocytes were isolated by in situ pre-perfusion and collagenase circulatory perfusion of rat livers. The effects of LPS associated with IFN-r, TNF-alpha and IL-1beta or IL-6 on NOS activity, cGMP, and NO(2)(-)+NO(3)(-) were observed in hepatocytes, respectively. Also the kinetic characteristics of this enzyme and dose response of corticosteroids on the induction of iNOS were analyzed. RESULTS: The maximum induction of NOS activity was observed in hepatocytes treated by LPS in combination with IFN-r, TNF-alpha and IL-1beta or IL-6. The kinetic analysis of this iNOS demonstrated specific constants of Km=10.8 micromol/L, Vmax=263.2 pmol/min/mg protein(for L-Arg), and Ki of 0.56, 0.94 micromol/L for competitive inhibitor, L-NMMA and NNA, respectively. The time course of induction showed that iNOS activity peaked at 9 h; however, significant increase in release of NO(2)( )+NO(3)(-) and cGMP sustained for at least 18 h. Dexamethasone and hydrocortisone dramatically inhibited the NOS induction in hepatocytes in vitro with IC50 of 3.5+/-10(-8)mol/L and 2.6+/-10(-6)mol/L, respectively. CONCLUSIONS: The expression of inductive NOS in hepatocytes requires specific synergetic action of cytokines, and the inducible characteristics may play an important pathogenesis in endotoxemia and septic shock. PMID- 11412396 TI - [The expression of uridine diphosphate glucuronosyltransferase mRNA regulated by alcohol]. AB - OBJECTIVE: To investigate whether ethanol and isopentanol could modulate the drug metabolism by regulating uridine diphosphate glucuronosyltransferase (UGT) expression in rat livers and cultured rat hepatocytes. METHODS: Rat livers and primary rat hepatocyte cultures have been used to determine the mRNA levels of five UGT isoenzymes following treatment with ethanol and isopentanol. RESULTS: Following treatment with alcohol in rats, UGT1A1 mRNA and UGT1A5 mRNA were increased to a mean of 177% and 166% of control, respectively. The mRNA levels of UGT2B1 and UGT2B3 were also increased in alcohol-treated rats to 178% and 132% of control, respectively. Incubation of hepatocytes with ethanol and isopentanol significantly increased the mRNA expression of UGT1A1, UGT1A5, UGT2B1 and UGT2B3 mRNA. CONCLUSIONS: The expression of UGT can be regulated by ethanol and isopentanol. As a result, chronic alcohol consumption may modify the metabolism of numerous endogenous and exogenous compounds, in particular some drugs, which are substrates of these UGT isoenzymes. PMID- 11412397 TI - [Influence of electroporation on the biological activities of primary rat hepatocytes]. AB - OBJECTIVE: To investigate the influence of electroporation on the biological activities of primary rat hepatocyte and to optimize the electroporation conditions introducing foreign genes into hepatocytes. METHODS: A single-pulse procedure was performed at low voltage (220-400 V) but high capacitance (500-950 microF). Its influence on hepatocyte activities was detected by Trypan blue exclusion (TBE) and MTT analysis. Besides, ALB, ALT and LDH in the supernatants of hepatocytes were tested by biochemical assay. RESULTS: Little hepatocyte damage and high survival rate (>90%) was found from 36 hours till 9th day of culture. At 36th hour after electroporation, ALB, ALT and LDH in the supernatants of Group B (220V, 950 microF) and C (400 V, 950 microF) were higher than those of control group. Whereas TBE and MTT analysis failed to indicate the significant difference of cell viability between electroporation groups and control group. CONCLUSIONS: This electroporation procedure is one of the optimal choices to introduce foreign genes into primary rat hepatocytes. PMID- 11412398 TI - [Non-Mendelian genetics and growth. The Russel-Silver syndrome]. PMID- 11412399 TI - [Incidence of IgE-mediated allergy to cow's milk proteins in the first year of life]. AB - OBJECTIVE: To study the incidence of IgE-mediated allergy to cow's milk proteins during the first year of life. METHODS: A multicenter, prospective study of newborns selected from different health centers was performed. The newborn infants were followed-up during the first year of life. Newborns with suspected adverse reaction to cow's milk were sent to the referral hospital for diagnostic study. This study was based on clinical history, skin tests (skin prick test) and on determination of specific IgE in serum (Pharmacia CAP system) against cow's milk and its protein fractions. Diagnosis was confirmed by open challenge. RESULTS: A total of 1,663 newborns were followed-up during the first year of life. Adverse reaction was suspected in 56 infants (3.3%). Allergy to cow's milk proteins was confirmed in 6 infants (0.36 %). Eighty-three percent of (5/6) children with cow's milk allergy had first-degree relatives with atopic disease compared with 19 % of children (329/1657) without cow's milk allergy. Among the entire sample, 26 infants had first-degree relatives with atopic disease and one of these infants (3.8%) developed cow milk allergy. The six children with cow's milk allergy were exclusively breast-fed, and clinical reaction developed within 1 week of the introduction of artificial feeding. CONCLUSION: The incidence of IgE-mediated allergy to cow's milk was 0.36 %. In infants with two first-degree family members with atopic disease, the probability of developing allergy to cow's milk proteins during the first year of life was 3.8%. PMID- 11412400 TI - [Bone mineral density determination by osteosonography in healthy children and adolescents: normal values]. AB - BACKGROUND: Osteoporosis is a frequent health problem in adults. Optimization of bone mass acquisition during childhood and adolescence may play a major role in the prevention of this disease. Osteosonography is a recent technique for measuring bone mineralization without exposing the patient to radiation. OBJECTIVES: To measure bone mineral density using osteosonography in healthy Spanish Caucasian children and adolescents in order to determine normal values. METHODS: We performed a cross sectional study of 829 healthy child and adolescent volunteers (360 girls and 469 boys) randomly selected from the urban area of Pamplona in Navarre (Spain). Ages ranged from 6 to 18 years. ADBM Sonic 1200 ultrasound densitometer from IGEA was used. Daily calcium dietary intake and amount of physical activity were recorded. RESULTS: Cross sectional standards for Ad-SOS are presented. Ad-SOS did not significantly change between the ages of 6 and 9 years in girls or until the age of 10 years in boys. From the ages of 10 to 14 years, Ad-SOS values were higher in girls than in boys. After the age of 14 years, no significant differences were found. No correlation was found between calcium dietary intake, amount of physical exercise or bone mineralization values. CONCLUSIONS: Measurement of Ad-SOS by osteosonography is an easy, fast and inexpensive method for measuring bone mineral density in children and adolescents without exposing them to radiation. It can be used in the pediatric population to detect early alterations in bone mineralization. PMID- 11412401 TI - [Energy and nutrient intake in compulsory high school students]. AB - OBJECTIVES: To analyze the dietary habits and energy and nutrient intake in secondary education students in a rural setting in order to design nutritional intervention strategies. MATERIAL AND METHODS: A semiquantitative food frequency questionnaire, stratified according to scholastic year, was randomly distributed to 406 adolescents (194 boys and 212 girls) aged between 13 and 16 years. RESULTS: Mean energy and nutrient intake was significantly higher in girls than in boys. In both sexes, 20.1% of energy was derived from meat, 17.7% from milk and milk products, 15.4% from cereals, 14.6% from sweets, 12.9% from vegetables, 7.2% from oils, 5% from fruits, 1.7% from eggs and 0.7% from fish. Protein accounted for 14.8% of energy intake, carbohydrates for 41.7%, total fat for 43.5%, and saturated fat for 16.5%. Cholesterol intake was excessive and over two thirds of protein intake was from animal sources. The mean intakes of calcium and magnesium, vitamins C and D and folate in boys, and the mean intakes of calcium, magnesium and iron, and vitamins C and D, riboflavin, niacin, vitamin B6 and folate in girls were below recommended levels. CONCLUSIONS: The dietary patterns of the secondary school students differed from the Mediterranean diet. Intakes of meat and sugar were too high, while that of cereals, legumes, fruits and fish were insufficient, leading to excessive fat intake from animal sources at the expense of complex carbohydrates and insufficient mineral and vitamin intake. Nutritional advice should be increased in primary care programs and nutritional education programs should be developed in secondary schools. PMID- 11412402 TI - [Hypercalcemia due to immobilization in critically ill children: calcitnonin treatment]. AB - OBJECTIVES: To evaluate the response to calcitonin treatment in critically ill children with hypercalcemia due to immobilization. PATIENTS AND METHODS: Four children aged 3-14 years were diagnosed with hypercalcemia due to immobilization. Detected levels of calcemia and ionic calcium ranged from 12.2-13.4 mg/dl and 1.44-2.2 mmol/l respectively. None of the children responded to rehydration or frusemide. Treatment was with subcutaneous and/or intranasal calcitonin at a dose of 6-20 IU /kg/day. RESULTS: After calcitonin administration, serum calcium decreased progressively until normal calcemia levels were reached between days 4 and 20 of treatment (8.6-10.5 mg/dl). No treatment-related adverse effects or alterations in phosphorus levels or alkaline phosphatases were observed. CONCLUSION: Calcitonin might be an effective treatment for hypercalcemia due to immobilization in critically ill children. PMID- 11412403 TI - [Relationship between respiratory syncytial virus subtype and clinical severity in bronchiolitis]. AB - BACKGROUND: Respiratory syncytial virus (RSV) is the most frequently implicated agent in bronchiolitis. The virus presents two antigenic subtypes (A and B). The results of studies on the relationship between viral subtype and clinical severity are conflicting. PURPOSE: To evaluate whether the subtype of this virus constitutes an independent variable in the severity of clinical expression of RSV infection. PATIENTS AND METHODS: We prospectively studied previously healthy children aged less than 2 years admitted for RSV infection during an epidemic season. Clinical presentation and evolution, laboratory and radiological aspects were compared in subsets of children infected with RSV subtype A or B. Clinical severity parameters included score and duration of respiratory distress, oxygen requirements, admission to the intensive care unit, mechanical ventilation and length of hospital stay. RESULTS: Eighty-five patients were enrolled in the study (50 boys and 35 girls; mean age 4.8,4.3 months). Fifty-nine children were infected with RSV subtype A. No differences were found in age, sex or antecedents but children with subtype B were less likely to have been breast-fed. Clinical presentation and laboratory and radiological findings were similar in both groups. No statistically significant differences in clinical severity parameters were found between subsets A and B. Age less than 3 months was associated with greater duration of respiratory distress and longer hospital stay. CONCLUSION: The results do not support the hypothesis that subtype A infection is associated with more severe respiratory syncytial virus disease. PMID- 11412404 TI - [Asthma and chronic diseases of the upper respiratory airway]. AB - There is enough evidence of an epidemiological, etiological and physiopathological link between chronic rhinitis (usually allergic) and asthma, and also between chronic sinusitis (generally bacterial) and this disease. Allergic inflammation of nasal mucosa could favor the development of chronic sinusitis. Indeed, some authors believe that the term "rhinosinusitis" should be used as nasal and sinus mucosa are one and the same thing. Some studies have demonstrated that treatment of chronic rhinitis or chronic sinusitis produces a benefit in the evolution of asthma when both diseases co-occur in the same patient, which suggests a therapeutic link. However, there is insufficient evidence that adenoidectomy and/or tonsillectomy worsen the evolution of asthma. When managing the asthmatic child, physicians should not forget to pay attention to the upper respiratory tract, evaluating and treating diseases in this area if necessary, as such therapeutic measures will also benefit asthma control. PMID- 11412405 TI - [Sudden infant death syndrome. Preventive environmental measures]. PMID- 11412406 TI - [Cystic fibrosis: nutritional considerations]. AB - During the last few decades, improved treatment measures and nutritional support in cystic fibrosis have increased survival and quality of life in these patients. There is a clear relationship between the development of malnutrition and worsening in respiratory function and both factors have been related to poor disease outcome. Malnutrition is a very frequent complication of this disease and is found in 20% of patients, due to negative energy-proteic balance. This disequilibrium can be explained by increased energy expenditure, high nutritional requirements and decreased oral intake. Periodic monitoring of clinical, anthropometrical and functional nutritional status is mandatory. Intake must be greater than requirements and specific nutritional support should be established when required. Patients with cystic fibrosis must receive a hypercaloric and hyperproteotic diet, with a high fat content, a normal quantity of carbohydrates and with pancreatic and liposoluble vitamin supplements in case of pancreatic insufficiency. PMID- 11412407 TI - [Chromosome aberrations in malformed newborns]. AB - OBJECTIVES: To determine the prevalence of chromosome abnormalities in malformed newborn infants and to analyze the distribution of the types of anomalies, and the variation in their frequency with maternal age. MATERIAL AND METHODS: We used the data collected according to the Spanish Collaborative Study of Congenital Malformations (ECEMC) methodology from March 1982 -September 1996. Of 33,562 newborns (live and stillborn), 1,409(4.1%) malformed infants were identified. A total of 332 karyotypes were performed in peripheral blood, representing 23.5% of the newborns with congenital defects. Results The frequency at birth of chromosome abnormalities was 5.4% of malformed newborns. There were 59 infants with Down's syndrome, 6 with trisomy 18, 3 with Turner's syndrome, 2 with trisomy 13, 2 with "Triple X", 1 tetraploidy, 1 triploidy, 1 trisomy 9 p, and 1 infant with a complex XXY mosaicism. The prevalence of Down's syndrome in the general population is 0.17%. The mean age of mothers with Down's syndrome infants was 34.2 years and paternal age was 36 years, and a non-statistically significant diminishing trend in mean maternal age was observed during the course of the study. The prevalence of Down's syndrome was higher in mothers aged more than 35 years. A non-statistically significant increase of the prevalence of Down's syndrome in newborns with mothers aged between 31 and 34 years was observed with time. The mean number of previous pregnancies was 2.81. Among a total of 49 mothers and fathers, two chromosome alterations were found. CONCLUSIONS: The prevalence of chromosome abnormalities in newborns with birth defects was 5.4%. The frequency of Down's syndrome was higher. Down's syndrome was more prevalent in mothers aged more than 35 years. The mean maternal age of Down's syndrome infants gradually diminished, and accumulated between the ages of 31 and 34 years. PMID- 11412408 TI - [Intrauterine dwarfism and dysmorfic features. A case of Russel-Silver syndrome]. AB - Russel-Silver syndrome is characterized by severe intrauterine growth retardation and is the most characteristic intrauterine dwarfism syndrome. In addition to short stature, low birth weight and reduced postnatal growth, this syndrome is characterized by features such a relative macrocephaly, a typical craniofacial appearance, asymmetry of the body and other abnormalities. Recent studies on developmental delay in these children have shown that most require special education. Attempts to explain the mechanism underlying this condition have been unsuccessful. Recent studies suggest a genetic cause, mainly uniparental disomy 7, although definitive data are lacking. We report a characteristic case of Russel-Silver syndrome: a newborn with fetal growth retardation, the craniofacial features described by Russel, relative macrocephaly, asymmetry of the body and very low weight increase. PMID- 11412409 TI - [Russel-Silver syndrome]. AB - Russel-Silver syndrome is a pattern of malformations whose most characteristic features are intrauterine and postnatal growth retardation, a characteristic facial appearance and limb asymmetry. We report a 9-month-old male born at 37 weeks' gestation. Family history, pregnancy and delivery revealed no significant anomalies. The infant had no intrauterine growth retardation (birth weight and height ranging between the 25 th and 50 th percentile). The infant's head circumference was disproportionately large for the small facial mass. Clinical findings included a small triangular face with frontal bossing, blue sclerae, thin lips with down-turned corners, retromicrognathia, clinodactyly of the fifth finger in both hands and limb asymmetry with undergrowth of the right leg and the left arm. The patient presented postnatal growth deficiency with weight and height 5th percentile; his head circumference grew normally along the 90 th percentile. Psychomotor development was appropriate for his age (9 months) according to the Sheridan Scale. On the basis of this clinical evidence we diagnosed Russel-Silver syndrome, attributing particular relevance to the normal intrauterine growth and the crossed asymmetry of the limbs. PMID- 11412410 TI - [Efficacy of lidocaine in neonatal ischemic ventricular tachycardia]. AB - A newborn diagnosed with D-transposition of the great arteries underwent an arterial switch operation when she was 7-days old. The coronary pattern was of type 2, with right coronary and circumflex arteries from the right sinus and anterior descending artery from the left sinus. Early post-repair electrocardiogram showed acute myocardial ischemia of the anterior septum and multiple episodes of sustained monomorphic ventricular tachycardia. Bolus and continuous infusion of intravenous lidocaine successfully terminated all episodes of ventricular tachycardia. At 6 months' follow-up the electrocardiogram was normal, without signs of myocardial ischemia or myocardial necrosis and without recurrence of ventricular tachycardia. PMID- 11412411 TI - [Albright hereditary osteodystrophy: identification of a novel mutation in a family]. AB - Studies to detect mutations in the GNAS1 gene were performed in a male patient with features of Albright hereditary osteodystrophy and resistance of target tissues to parathyroid hormone (Pseudohypoparathyroidism Ia). The same investigations were carried out in the patient's mother who showed somatic features of Albright's hereditary osteodystrophy and brachymetacarpia without resistance to parathyroid hormone (Pseudopseudohypoparathyroidism). A point mutation designated c.794GA (R265H) in exon 10 of GNAS1 was identified in DNA from the patient and his mother. This novel mutation in exon 10 of GNA PMID- 11412412 TI - [Alice in Wonderland syndrome due to Epstein-Barr virus infection]. AB - The Alice in Wonderland syndrome refers to distortions in body image and in the apparent sizes, shapes, and spatial relations of objects seen. The syndrome is usually associated with migraine headaches and has also been reported in several viral infections. We report a 6-year-old boy who presented to the emergency department complaining of several episodes in which the ceiling, the objects and the people around him seemed very small and far away. The child presented no alteration in the level of consciousness. The episodes provoked great fear in the child. Physical examination revealed no abnormalities except pharyngoamygdalitis. Serologic studies (IgM antibodies to viral capsid antigen) confirmed Epstein-Barr virus infection. The child's symptoms resolved spontaneously within 48 hours and he continued to be asymptomatic after a 4 -month follow-up. We consider that all children presenting a clinical picture consistent with the Alice in Wonderland syndrome should undergo serological testing for Epstein Barr virus infection. Diagnosis would enable physicians to reassure the family of the temporary and benign nature of this alarming condition. PMID- 11412413 TI - [Generalized petechial exanthema]. PMID- 11412414 TI - [Critical evaluation of original articles published in Anales Espanoles de Pediatria 1999-2000]. PMID- 11412415 TI - [Law of minors and public disorder]. PMID- 11412416 TI - [Current use of antipyretics in children at home]. PMID- 11412417 TI - [Seroepidemiological study of the measles virus]. PMID- 11412418 TI - [Extrusion of a ventriculoperitoneal shunt catheter through the mouth]. PMID- 11412419 TI - [Pleural effusion associated with hepatitis A]. PMID- 11412420 TI - [Excuse me doctor, but this baby smells of licorice]. PMID- 11412421 TI - [Megabladder-microcolon-intestinal hypoperistalsis syndrome]. PMID- 11412460 TI - [Acute gastroenteritis:oral rehydration and nutritional treatment. Are we doing what should be doing?]. PMID- 11412461 TI - [The young febrile child. Results of a multicenter survey]. AB - OBJECTIVES: To establish the value of clinical and laboratory parameters gathered from a national survey on the management of the young febrile child (0-36 months) in emergency departments and to determine the etiology of potentially serious bacterial infections in Spain in this group of patients. PATIENTS AND METHODS: Prospective survey of febrile children aged less than 36 months who presented to the emergency departments of the participant hospitals and who fulfilled the inclusion criteria. RESULTS: A total of 937 cases were collected. Of these, 43 were excluded because of absence of blood culture and 151 because the patients had not undergone obligatory monitoring until they became afebrile or had no final diagnosis. Consequently, the sample size consisted of 743 patients. Clinical assessment was very important to detect patients with potentially serious bacterial disease. Among the laboratory parameters, a white blood cell count above 15,000/l or below 5,000/l and elevated C-reactive protein levels were the most useful. Urinary tract infection was the most frequent invasive disease. Neisseria meningitidis was the most frequent microorganism isolated in the blood culture. The incidence of occult bacteremia was 3 % with predominance of Streptococcus pneumoniae (74 %). CONCLUSIONS: The experience of the pediatrician supported by clinical and laboratory parameters are the most effective means of detecting febrile children (0-36 months) with potentially serious bacterial disease. PMID- 11412462 TI - [Enteroviral meningitis. Clinical and laboratory findings in a series of 60 children]. AB - OBJECTIVE: To describe the epidemiologic, clinical and laboratory findings in a series of children with enteroviral meningitis. PATIENTS AND METHODS: We performed a retrospective descriptive study of all the children admitted to our hospital between May 1996 and September 1997 with a diagnosis of enteroviral meningitis, defined as either a positive cerebrospinal fluid (CSF) culture for enterovirus or pleocytosis in conjunction with a positive enterovirus culture at any other site (rectal or pharyngeal swabs). RESULTS: Sixty cases of enteroviral meningitis were found, all of which occurred between May and July. The median age was 6 years. Half the patients were diagnosed in the first 6 hours of illness and 20 % in the first 2 hours. The most frequent symptoms were headache (97 %) and vomiting (80 %). Fever was present in only 56 % of the patients. On physical examination 78 % presented positive meningeal signs. A total white blood cell count of more than 15,000/l was present in 28 %. CSF white cell count was over 500/l in only five patients and in four pleocytosis was absent; a predominance of polymorphonuclear cells was found in 45 %. Enterovirus was isolated 43 % of the CSF specimens. The median length of hospital stay was 2 days. Antibiotic treatment was started in 13 % of the patients and all recovered without sequelae. CONCLUSIONS: All the cases occurred in late spring. Fever was an inconstant finding. Laboratory findings were typical of viral meningitis, although the percentage of neutrophils in CSF varied widely, with more than 50 % of neutrophils in half of the patients; the absence of pleocytosis was not unusual. The enterovirus yield from the CSF specimens was higher than in previously reported in Spanish series but did not influence the management of our patients. PMID- 11412463 TI - [Utility of Boyer's score modified for the differential diagnosis of bacterial and viral meningitis]. AB - BACKGROUND: Many studies have been performed to establish criteria for the differential diagnosis between bacterial and viral meningitis. In 1980, Thome and Boyer proposed a score that has been widely used. OBJECTIVES: The aim of this study was to assess the utility of this score when applied to our patients with meningitis and to evaluate the diagnostic yield after adding other laboratory tests. METHODS: We retrospectively studied the children diagnosed with meningitis in the Hospital Nino Jesus between January 1993 and February 2000. In all patients, Boyer's score and the following laboratory variables were applied: reactive C protein levels, percentage of neutrophils in the cerebrospinal fluid depending on age, and the ratio of immature cells/total neutrophils in the blood. We calculated the sensitivity and specificity of Boyer's score and the laboratory variables. RESULTS: Of the 476 children with meningitis, 402 had viral meningitis and 74 had bacterial meningitis. All the children with bacterial meningitis except seven had a score higher than 3 (a doubtful indication for antibiotic treatment). The sensitivity and specificity of Boyer's score was 90 % and 99 % respectively. When laboratory variables were added, sensitivity was 100 % and specificity was 98 %. CONCLUSIONS: Boyer's score is a useful tool for the differential diagnosis of meningitis, but its diagnostic yield is considerably increased when the results of laboratory tests are added. PMID- 11412464 TI - [Acute osteomyelitis. Clinical, radiological and bacteriological features and outcome]. AB - OBJECTIVE: To compare the clinical, radiological, bacteriological features and outcome of neonatal patients with acute osteomyelitis aged (28 days with those of children aged <= 28 days of age. PATIENTS AND METHODS: A retrospective analysis was performed. The clinical histories of patients aged less than 15 years diagnosed with acute osteomyelitis in a tertiary care hospital were reviewed. Acute osteomyelitis was defined as the association between at least two of the following variables: a) positive blood or biopsy culture; b) purulent bone aspiration; c) clinical features compatible with a diagnosis of acute osteomyelitis; d) radiological features suggestive of acute osteomyelitis, and e) favorable outcome after antibiotic treatment. The patients were divided into two groups: group 1: neonatal patients (aged (28 days) and group 2: children aged <= 28 days. Statistical analysis was performed by the Chi-square test and Student's t-test for categorized and numeric variables, respectively. RESULTS: Between 1977 and 1999, 45 children aged less than 15 years old were diagnosed with acute osteomyelitis. Of these, 17 patients were neonates (group 1) and 28 patients were aged more than 28 days old (group 2). The male:female ratio was 1.1:1 in group 1 and 3:1 in group 2. The mean age was 17.7 +/- 7.5 days in group 1 and 7.2 +/- 4.3 years in group 2. Metaphysis and tibia were more commonly affected in group 2 (p < 0.05). Epiphysis, arthritis and humerus were more frequently affected in group 1 (p < 0.05). Osteolysis and periosteal reaction predominated in group 1 (p < 0.05). 99mTc Bone scintigraphy and magnetic resonance imaging showed pathological findings in all patients. Blood culture revealed Staphylococcus aureus in 46 % of the patients. Blood and biopsy material cultures were positive in 46 % and 75 %, respectively. Of the 45 patients, outcome was favorable in 37 (82.2 %). CONCLUSION: Acute osteomyelitis showed different characteristics in the neonatal and postnatal periods. Bone scintigraphy and magnetic resonance showed high sensitivity. Bacteriology was positive in 50 % of patients. PMID- 11412465 TI - [Epidemic outbreak of tuberculosis]. AB - OBJECTIVE: To describe how collaboration between epidemiological surveillance and primary and specialist health care centres enabled rapid intervention during an outbreak of tuberculosis in a nursery school. MATERIAL AND METHODS: A child minder was diagnosed with tuberculosis. The persons exposed were identified. These were four child minders and 58 children under 4 years of age. The respective primary care pediatricians carried out a tuberculin test in all of the children. Children with a positive tuberculin test underwent radiological and microbiological study to rule out the disease. The adults underwent the same procedure. RESULTS: Detection among the children was rapid; 32.8 % were infected and six showed alterations in thoracic x-rays and were considered to be ill. In three children Mycobacterium tuberculosis was isolated and was genetically similar to the index case. Primary chemoprophylaxis was carried out in all children with a negative tuberculin test; secondary chemoprophylaxis was administered to infected children and specific treatment to the ill. In all children, evolution was satisfactory. CONCLUSIONS: Periodic surveillance for tuberculosis should be carried out among adults working with children. Genetic study of the strains isolated facilitates epidemiological analysis of these microepidemics. PMID- 11412466 TI - [Childhood immunization schedule 2001-2002. Advisory Committee on Vaccines of the Spanish Association of Pediatrics]. AB - In 1994 the Spanish Association of Pediatrics founded the Advisory Committee on Vaccines with the aim of providing advice on matters related to childhood immunizations and of implementing vaccination schedules. The latest recommendations concern the immunization schedule for 2001-2002, in which indications for the inactivated poliovirus vaccine instead of the attenuated poliovirus vaccine are of prime importance. The advisability of including the vaccine against chicken pox in healthy children is stressed. PMID- 11412467 TI - [Active immune-prophylaxis after allogeneic hematopoietic progenitor cell transplantation in pediatric patients]. AB - There is considerable evidence supporting the disappearance of immunity against diseases for which there is active immune-prophylaxis available after allogeneic hematopoietic progenitor cell transplantation. These findings do not constitute merely an epidemiologic problem. From a strictly clinical point of view, these patients might benefit from the administration of different forms of vaccines against diseases to which they are particularly susceptible. There are, however, no unified criteria for the revaccination of these patients. Moreover, several surveys suggest that active immune-prophylaxis is frequently undervalued in these patients. In the present review, we provide up-to-date information on current trends as well as practical guidelines for the re-immunization of these patients in our environment. PMID- 11412468 TI - [Cytogenetic abnormalities in acute lymphoblastic leukemia]. AB - Cytogenetic analysis of blast cells in childhood acute lymphoblastic leukemia has led to the recognition of specific non-random chromosomal abnormalities with prognostic value. Most patients with ALL show karyotype abnormalities, either in chromosome number (ploidy) or as structural changes such as translocations, inversions, or deletions. Many of these chromosomal alterations are associated with specific cytomorphological and immunological types. The greatest impact on patient management has been the finding that the cytogenetic result is an independent prognostic indicator. Certain karyotypes are associated with a favorable prognosis while others indicate a poor outcome. This has led to the administration of alternative therapies according to risk. For instance, hyperdiploidy with a modal chromosome number of 51 or greater, which represents 25-30 % of all cases of ALL, has proved to have the most favorable prognosis among established ploidy groups, whilst translocations such as the Philadelphia translocation t(9;22) and t(4;11) are associated with a poor prognosis. This study focuses on the most important chromosomal abnormalities found in childhood ALL and their prognostic and therapeutic implications. PMID- 11412469 TI - [Impact of new eligibility criteria on a screening program for retinopathy of prematurity. one-year evaluation]. AB - OBJECTIVE: To evaluate, in clinical and economical terms, a screening program for retinopathy of prematurity 1 year after implementing new eligibility criteria. METHOD: New criteria to safely reduce the number of newborn infants screened without missing any stage 3 retinopathies of prematurity were established after reviewing the clinical information from the 1991-1998 screening program. In 1999 the infants were screened according to the new eligibility criteria. The absolute and relative reduction in the number of patients requiring screening was determined by comparing the implementation of the new eligibility criteria with the previous criteria. Criteria 1991-1998: a) Birth weight 1,500 g or gestational age 33 weeks; and b) gestational age 34-36 weeks with exposure to supplemental oxygen for at least 24 h. New criteria 1999: a) Birth weight 1,250 g or gestational age 30 weeks; and b) infants with birth weight > 1,250 g or gestational age > 30 weeks with very serious neonatal medical problems according to the neonatologist. RESULTS: In 1999, 244 patients would have been screened with the 1991-1998 criteria. Applying the new 1999 criteria, only 72 patients were screened. The number of examinations saved was 369 and the cost of the program decreased by 30,000 euros. CONCLUSIONS: The new eligibility criteria for retinopathy of prematurity screening has reduced the number of patients screened by 70 %. To ensure inclusion of all newborn infants at risk of developing the disease and to minimize the number of screened infants and cost of the program, screening strategies can be tailored to health centers' specific characteristics. PMID- 11412470 TI - [Neonatal varicella: report of a case of bronchopneumonia and hemorrhagic pulmonary edema]. AB - We describe the case of a male newborn infant, whose mother developed varicella 20 days before delivery. At 2 hours of life the infant's general state suddenly deteriorated and he presented cyanosis, respiratory distress and emission of pink foam in his mouth. The infant was diagnosed with bilateral bronchopneumonia, of probable varicellosus etiology, with a component of hemorrhagic pulmonary edema. He required mechanical ventilation and inotropic support for 48 hours. Response to endovenous acyclovir was favorable. At 10 days of life varicella-zoster virus (VZV) in cerebrospinal fluid was detected by polymerase chain reaction and seroconversion of IgG anti-VZV was positive. PMID- 11412471 TI - [Partial trisomy 18q due to maternal reciprocal translocation 4;18]. AB - We report a new case of partial trisomy 18q due to a balanced reciprocal translocation 4;18 in the mother. The female infant had a partial trisomy of the long arm of chromosome 18 associated with a partial monosomy of distal 4q. The infant showed many of the main clinical features of trisomy 18, such as dysmorphic face, congenital heart defect, crossing of the second and fifth fingers over the third and fourth with flexion contractures, and abnormal genitalia. We believe that the trisomy 18 phenotype requires a large region of 18q and that the greater the trisomic fragment, the more severe the expression. We stress the importance of genetic counseling to carriers of balanced translocations. The risk for each case should be evaluated and information should be given on the possibility of prenatal diagnosis. PMID- 11412472 TI - [Necrotic ulcers in a child with fever]. PMID- 11412473 TI - [Ocular infiltration in the anterior chamber in a female infant with acute non lymphoblastic leukemia]. AB - Unlike myeloid sarcoma, ocular involvement is unusual in acute non-lymphoblastic leukemia.A 9-month-old female infant with acute non-lymphoblastic leukemia M5 and evidence of active central nervous system (CNS) disease showed infiltration of the anterior chamber during therapy. At that time, the CNS disease was in completed remission. She was treated with topical corticosteroids, chemotherapy and bilateral ocular radiotherapy (total dose 1,000 cGy). The ocular manifestations responded well to treatment but hematologic response was poor. The patient died a few months later. Any ophthalmic manifestation in children with leukemia should be detected and treated early. Radiotherapy is warranted in infiltration of the anterior chamber of the eye. The presence of ocular, central CNS or bone marrow involvement indicates poor prognosis in acute childhood leukemia. PMID- 11412474 TI - [Arterial hypertension, hematuria and renal involvement before chickenpox]. AB - Chickenpox in childhood is usually a self-limiting disease with relatively few and benign complications, although serious complications do occur. Renal involvement is uncommon during the course of this disease. We describe a 6-year old girl who developed clinical glomerulonephritis, with arterial hypertension, hematuria and transient minor disturbance of renal function revealed by creatinine clearance 2 days before the appearance of papulovesicular skin lesions characteristic of chickenpox. The patient recovered spontaneously in 2 weeks. PMID- 11412475 TI - [Pleurodesis with tetracyclines after fontan operation]. AB - Chemical pleurodesis consists of the fusion of both pleural layers after contact with certain substances, and is used in the treatment of prolonged or recurrent pleural effusions refractory to other therapeutic approaches. This procedure has been extensively used in adults with malignant pleural effusion but has rarely been used in children. The Fontan technique is used in the treatment of tricuspid atresia and other heart diseases that act as a single ventricle. This technique frequently presents complications due to fluid retention (edema, ascites, pleural and pericardial effusions and protein-losing enteropathy). With persistent or recurrent pleural effusion, chemical pleurodesis through the drainage tube might constitute a safe and reliable alternative treatment. We report the case of a female patient with tricuspid atresia, who presented recurrent pleural effusion after undergoing the Fontan procedure. The disorder was resolved by chemical pleurodesis with tetracycline. PMID- 11412476 TI - [Myxoma of the left ventricular outflow tract]. AB - We report the case of a 5 year-old girl who presented asymptomatic myxoma in the left ventricular outflow tract. The tumor was initially detected by echocardiography, which was performed for heart murmur. This diagnosis was later confirmed by magnetic angioresonance imaging which enabled prompt surgical removal. We emphasize the advantages of transthoracic echocardiography in the early diagnosis of this kind of tumor. PMID- 11412477 TI - [Systemic myofibromatosis in an infant]. AB - Infantile myofibromatosis is characterized by the presence of solitary or multicentric fibrous nodules in skin, muscle and/or internal organs. Despite being an infrequent entity, and consequently little known by pediatricians, it constitutes the most frequent fibrous tumor in children. Solitary cutaneous lesions have a very good prognosis but in some cases there is systemic involvement. In these cases the disease can produce serious complications and even put the patient's life at risk, especially during the first months of life. These complications are due to the locally invasive nature of the nodules, obstruction of vital organs, growth retardation or infection. We present the case of an infant who at birth presented a skin nodule only. Over time, the infant presented lesions in skin, bone and internal organs. The infant showed respiratory distress requiring mechanical ventilation due to diaphragmatic paralysis. We recommend close follow-up of all patients with infantile myofibromatosis to prevent or make an early diagnosis of these complications PMID- 11412478 TI - [Inner ear autoimmune disorder. Cogan's syndrome]. AB - We present a case of Cogan's syndrome (interstitial keratitis, rapidly evolving sensorineural hearing loss and vestibular dysfunction) in a 12-year-old girl. Diagnosis was made by clinical and audiologic examination as well as by nonspecific laboratory findings (increased erythrosedimentation rate and positive IgA and IgM rheumatoid factor titers and antineutrophil cytoplasmic autoantibodies). Detection of specific autoantibodies against cochlear antigenic epitopes with a molecular weight of 68 kDa by Western-blot assay confirmed an organ-specific autoimmune mechanism. Therapy with deflazacort 1 mg/kg per day 3 three weeks decreasing progressively thereafter was successful and the patient remains symptom-free. PMID- 11412479 TI - [Poisoning after the use of sodium phosphate enema]. PMID- 11412480 TI - [Accidents with supermarket carts as a cause of childhood hospitalization]. PMID- 11412481 TI - [Smith-Lemli-Opitz syndrome]. PMID- 11412482 TI - [Ring 13 chromosome]. PMID- 11412483 TI - [Monosomy r(13), a new observation]. PMID- 11412487 TI - [Tridimensional theory of personality: applications to substance abuse disorders]. AB - INTRODUCTION: We have assessed the weight that Cloninger' dimensions play in the substance abuse disorder. Also, we have analysed the hypothetical self independence of these dimensions and if there is some correlation between those and some demographic variables. METHODOLOGY: 20 drug abstinents and 20 controls, all males, fulfilled the Tridimensional Personality Questionnaire. RESULTS: All the values on the Novelty Seeking (NS) subclass and the overall NS scale were higher in the abstinent group. The most meaningful differences were found on the excitable and extravagant subclass and the total NS scale. There was also a significant though smaller difference on the disorderly subclass. Only the Harm Avoidance (HA) subclass fatigability was significantly higher in the abstinent group. The value of persistence substantially low in the abstinent subjects was the most significant difference between groups when the Reward Dependence (RD) scale was considered. The correlation analysis demonstrated that the three dimensions were mutually independent in the controls. However, in the abstinent group NS correlated positively with HA and negatively with persistence. Any correlation was found between the dimensions and the demographic variables in both groups. CONCLUSION: A high sensation seeking behaviour and a low persistence seems to be the most prominent characteristic of our abstinent subjects. Both tendencies could explain in part their high substance seeking tendency and possibly the high rate of relapse found in similar populations. Finally, the Cloninger's hypothesis about the mutually independence of the dimensions seems to be fulfilled only in the control group. PMID- 11412488 TI - [Effectiveness and safety of topiramate in treatment-resistant bipolar disorder]. AB - INTRODUCTION: This study was conducted to evaluate the effectiveness and safety of topiramate as add-on therapy for treatment-resistant bipolar disorder. METHODS: Twenty-one DSM-IV bipolar patients, considered resistant to treatment with lithium, carbamazepine or valproate, gave informed consent to receive increasing doses of the novel anticonvulsant topiramate as adjunctive therapy for their manic (n= 9), depressive (n= 6), hypomanic (n= 3), mixed (n= 2) or schizoaffective manic (n= 1) symptoms. The dosage of other mood stabilizer drugs remained unchanged throughout the 6-week follow-up. Outcome measures included the YMRS, HDRS-17, and CGI scales. Fifteen out of 21 patients completed the 6-week follow-up. RESULTS: Six patients (40% of completers, 29% by intention-to-treat) were considered responders to topiramate (> 50% reduction in YMRS or HDRS-17 and a decrease of 2 points in CGI). The drug was less effective in intially depressed patients. Topiramate was well tolerated and only one patient discontinued due to side-effects. The most common adverse effect was paresthesia (n= 2). Ten patients experienced moderate weight loss during the follow-up. The mean topiramate dose at endpoint was 158 mg/day. CONCLUSIONS: These preliminary results suggest that topiramate may be a useful therapy for bipolar disorders, with promising results even in the most treatment-refractory patients. PMID- 11412489 TI - [Factors associated to non-attendance to psychiatric first visit]. AB - INTRODUCTION: The failure to the first regular appointments in Mental Health Centres represent a problem in economic terms for the institution, a waste of time for the professional person and an increase in the waiting list in the centre with the subsequent prejudice for the patient. METHODOLOGY: In this investigation we study the influence of some sociodemographic (sex and age) and referral variables (geographic area, previous contact in the centre, referral reason and diagnostic by the family physician) in a group of 235 failing subjects compared with a group of 300 attending subjects. RESULTS: Results show that there's no relationship between the variables in terms of sex or age; that there are differences related to the geographic area from where they are referred and that the patients with a previous contact in the centre (more than one year) fail less than those who never attended. PMID- 11412490 TI - [Delirium in a consultation liaison psychiatry unit of a general hospital]. AB - INTRODUCTION: Delirium is an organic mental disorder defined as a confusional state, attention deficit and disorganized thinking, with a fluctuating course and acute development. Underlying causes are numerous, but the most common are multiple medical problems and drug side effects. SUBJECTS AND METHOD: We present a descriptive and transversal study of 62 inpatients with delirium diagnosis (ICD 10 criteria) attended by a Liaison Psychiatric Department in a General Hospital during a year. The data included demographic, somatic and psychiatric variables. RESULTS: The overall referral rate from the general hospital was 0.2%. No significant differences, were found in sex distribution. The age average was 64.46 years. The majority of patients were referred by Surgery (19.4%) and Medicine (17.7%). The most common medical diagnosis was cancer (25%) and respiratory failure (15.9%). Haloperidol was the most frequent psychiatric treatment. The greatest number of referred patients were included in the serious medical state and the 23.9% of patients with delirium diagnosis deceased. CONCLUSION: Data supports the statement that delirium is most often found in hospitalized patients with a diagnosis of cancer and respiratory failure, and that this diagnosis is associated with mortality in a high proportion. Furthermore, this study suggest an increase in cost-containment since the average stay are increased in comparison with the rest of hospitalized patients. PMID- 11412491 TI - [Comparative study of the needs of patients with schizophrenia by gender]. AB - OBJECTIVES: To evaluate the gender differences in the needs of the people with schizophrenia who live in the community. METHOD: We randomly selected 231 patients with schizophrenia who were receiving treatment in one of the five mental health care centres that participated in the study. Patients were evaluated with a socio-demographic questionnaire, the Positive and Negative Syndromes Scale (PANSS) and Camberwell Assessment of Need (CAN) questionnaire. The CAN evaluate 22 needs, that we could be grouped into five subscales: service needs, basic needs, functioning need, health needs and social needs. All needs were evaluated by the professional and the user independently. RESULTS: Professionals detected more needs in men than in women, but users did not report such difference. Both professionals and users detected more service needs for women (p< 0.005) and more basic needs for men (p< 0,01-0.005). Only professionals detected more functioning needs in men (p< 0.05). CONCLUSIONS: Professionals detect more needs than users and detect more needs for men than for women. Women have less needs in food, personal care, home care, and daily activities. These gender differences should be taken into account when designing treatment programs for schizophrenia. PMID- 11412492 TI - [Antipsychotics use in drug abuse]. AB - A review was carried out into the literature of the last ten (Medline, IME, HealthSTAR, Cinahl) on the use of antipsychotic drugs in substance abusers. The use of these drugs is common in this population because of the high prevalence of psychotic pathologies among substance abusers and they are used in the treatment of situations derived from the consumption of certain substances and in secondary behavioural disorders following organic mental disorders or personality disorders. The special characteristics for the use of these drugs among addicts are reviewed as these patients tend to present adverse side-effects with greater frequency, for example, extrapyramidal symptoms and less compliance with treatment. PMID- 11412493 TI - [Serotonin receptor changes in depression: evidences and limitations]. AB - Serotonin and serotonin receptors might be involved in the pathophysiology of depression. In the following, research data supporting the general hypothesis of adaptatives changes in density and functioning of serotonergic receptors in depression are review. Binding assays, platelet and neuroendocrine studies supports this theory. The density of 5-HT2A binding sites in postmortem brain tissue of depressed patients and suicide victims, as well as in platelets of drug free depressed patients has been found to be increased by several authors. The reduce hormonal response to fenfluramine challenge test in depression appears to indicate a sub-normal functioning of 5-HT2A receptors, however studies evaluating physiologic platelet 5-HT2A receptor-mediated responses have produced conflicting results. On the other hand, neuroendocrine challenges tests with 5-HT1A agonists suggest that presynaptic and postsynaptic 5-HT1A receptors may be also desensitized in depression. To date, postmorten receptor 5-HT1A studies in suicide victims have not yielded consistent. Taken together, these findings provide support for hypotheses of amine receptor abnormalities in depression, and indicate the need for expanded studies of amine receptor density and function in depression. Nevertheless, the role of these changes in the pathophysiology of depression has not been proved. PMID- 11412494 TI - [Critical review of conceptual aspects on social phobia]. AB - Two important nosological areas of debate currently exist concerning social phobia. The first focuses on the denomination of this disorder. In this respect, experts recommend a change in the denomination to Social Anxiety Disorder, since this better expresses the magnitude and the impact of the illness and focuses less on the behavioral aspect (avoidance) of the disorder. The second area refers to the current situation of overlap between social phobia and avoidance personality disorder. Experts on this field consider that avoidance personality disorder represents a severe form of generalised social phobia, and therefore, that it should disappear as an independent diagnostical category on Axis II, and become part of social phobia on axis I. PMID- 11412495 TI - [Mechanism of action of electroconvulsive therapy in depression]. AB - The knowledge of the therapeutic efficacy of the electroconvulsive therapy (ECT) dates back to the fourties. Since then important investigation efforts have been carried out, to analyze the mechanism of action by which the ECT achieves its therapeutic effect. The present work reviews the different hypothesis that have been related to the mechanism of action or the ECT, as well as the large neurophysiological findings, believed to be involved in its therapeutic effect. PMID- 11412496 TI - [Psychotic disorder in Prader Willi syndrome: a case report]. AB - In the Prader-Willi Syndrome the psychiatric symptoms are fundamentally behavioral disorders and obsessive-compulsive symptoms in relation with food. These aspects have been published previously. Nevertheless, psychotic manifestations in this syndrome are infrequent. Let us present a case of this comorbidity and a bibliographic review in last years (Medline, EmBase, Cochrane) was done about this association. PMID- 11412498 TI - [Hospital-at-home, a new health care approach?]. PMID- 11412499 TI - [Surgery and combined therapy for non-small cell lung cancer with invasion of the mediastinal nodes. A retrospective study]. AB - OBJECTIVE: To analyze the survival of patients classified N2M0 (N2 cytology/histology)with non-small cell lung cancer treated by surgical resection of the primary tumor, lymphadenectomy and neo-adjuvant therapy. PATIENTS AND METHODS: Among 1,043 consecutive patients with lung cancer treated between 1990 and 2000, 155 were classified N2M0 by histology. Of 130 patients undergoing thoracotomy, excision of the primary pulmonary tumor and lymphadenectomy were performed in 116. Among the 116 N2M0patients undergoing surgical resection, 23 were diagnosed N2c(c3)by mediastinoscopy and/or mediastinotomy and received induction chemotherapy (CT) with mitomycin/ifosfamide/cisplatin (3 cycles)and 93 were diagnosed N2pM0 after examination of samples of mediastinal lymph tissue taken during thoracotomy; for 19 of these patients,earlier surgical exploration of the mediastinum had been negative. The patient diagnosed N2p after thoracotomy also received CT and/or radiotherapy (RT). N2p patients who received induction CT also received RT. Those who were negative after lymphadenectomy and severely ill patients received no adjuvant therapy of any type. RESULTS: Mean survival of resected patients (23/49) diagnosed N2(C3) by mediastinoscopy/mediastinotomy and who received induction CT was 18 months. Survival at 1, 2 and 5 years was 80%, 45% and 30%, respectively. No postoperative deaths occurred in this group. One patient developed a bronchopleural fistula. Nine patients showed no signs of residual mediastinal node disease after lymphadenectomy. The mean survival of resected patients (93/106) diagnosed N2p after thoracotomy was 13 months and survival rates at 1, 2 and 5 years were 56%, 31% and 19%,respectively. Fourteen patients in this group died within 30 days of surgery. Nine patient developed bronchopleural fistulas. The difference in survival between the two groups was not significant. CONCLUSIONS: Histologic or cytologic confirmation of N2 disease can be considered to indicate poor prognosis. Standard, complete surgery with induction CT in selected patients improves survival for those diagnosed N2 upon thoracotomy, with no statistically significant differences. PMID- 11412500 TI - [Prognostic factors and outcome of thymectomy in 80 cases of myasthenia gravis]. AB - OBJECTIVES: To analyze the results of thymectomy in our series of patients with myasthenia gravis (MG) and to study the influence of the most common prognostic factors. MATERIAL AND METHODS: Eighty MG patients over a period of 23 years underwent thymectomy consecutively in our hospital. Preoperative assessment included clinical evaluation of muscle weakness, edrophonium testing, electromyography, lung function testing, chest X-rays and CAT scans. Symptoms were assessed by the Osserman scale. The surgical approach was amplified transsternal thymectomy. The prognostic factors studied were sex, age, clinical stage, duration of disease before surgery and histology of the thymus. Clinical outcome was assessed using Millichap and Dodge's criteria. Follow-up was by the chest surgery and neurology departments. RESULTS: Complete remission was observed in 29 cases (36.2%) and significant improvement in 42 (52.5%). Complications developed in 9 patients (11.2%). Most patients were women (53/27) and outcomes for men and women were not statistically different. Mean age was 36 years (range 11-79), with no significant difference in outcome for patients who were older or younger than 60 years of age. Nor were differences evident related to presurgical clinical stage or levels of severity (I + IIa/IIb + III). Differences in outcome were highly significantly related to duration of disease (< 24 / > 24 months) (p = 0.0022), such that outcome was more satisfactory when the pre-surgical course of disease was shorter, provided that no thymoma was present. CONCLUSIONS: Amplified transsternal thymectomy was safe and effective for those patients with MG. When disease had been present for less than two years, the prognosis was better. PMID- 11412501 TI - [Gain, loss and agreement between respiratory specialists and generalists in the diagnosis of asthma]. AB - OBJECTIVE: To determine and analyze the degree of agreement and disagreement in the diagnosis of bronchial asthma (BA) by respiratory disease specialists and generalists in regional hospital and primary care settings. MATERIAL AND METHODS: Ninety-six outpatients (16 to 70 years of age) were studied; all had been assigned a diagnosis of BA by the referring physician or by the respiratory disease specialist. We recorded 1) clinical symptoms, determining the initial probability of a diagnosis (IPD)of BA to be high, medium or low; 2) results of spirometry and bronchodilator testing (BDT), peak flow variability and methacholine challenge testing; 3) prick test results, eosinophil levels and total serum IgE levels. Three diagnoses were recorded: the initial diagnosis (ID) by the referring physician to whom follow-up data were unavailable; diagnosis by the respiratory disease specialist based only on clinical symptoms (RSS); and the final diagnosis(FD). To arrive at a FD of BA, it was necessary to have a high or medium IPD and a positive BDT. A Kappa test was used to analyze the degree of agreement among the three diagnoses. Group features associated with greater or lesser agreement were analyzed by chi-square tests and analysis of variance. RESULTS: Agreement was acceptable between RSS and FD (K = 0.63) but very low between ID and RSS and between ID and FD. In the latter two cases, agreement was greatest for patients diagnosed in hospital and for those with high IgE levels (p < 0.05), with high IPD, longer course of disease and a history of asthma (p < 0.01) (odds ratio =59.8). Diagnostic disagreement occurred mainly for patients for whom a BA diagnosis was gained later, the of under-diagnosis being 39%. The patients involved visited the physician only because they had observed an isolated symptom related to asthma (odds ratio = 119) and to arrive at a diagnosis bronchomotor tests other than BDT were required (p < 0.01). CONCLUSIONS: a) The degree of agreement for a diagnosis of BA is low. b)The functional profile of patients for whom diagnostic agreement exists differs from that of patients for whom diagnosis is gained through testing. c) In the context of this study, a high rate of under-diagnosis is evident. PMID- 11412502 TI - [Outcome of treating initial tuberculosis in the area south of Seville in a 5 year period (1994-1998]. AB - OBJECTIVE: To analyze the outcome of treating initial tuberculosis (ITB) in the hospital district south of Seville (Spain). MATERIAL AND METHOD: A descriptive study of treatment outcome in cases of ITB in our district between 1 January 1994 and 31 December 1998. Outcome was classified according to World Health Organization guidelines:successful (S) potentially unsuccessful (PU), exitus (EX) and transferred out (T).Four hundred fifty-four patients (mean age 35 years) were enrolled; 22% were HIV+ and 21% were intravenous drug users (IVDU). Treatment was self-administered in all cases, with 98% being given the standard prescription of 6 or 9 drugs. No subsequent case history was found for 15 cases, such that outcome could not be assessed. RESULTS: Of the 439 evaluable cases, outcome was S for 74% (326/439), PU for 16% (70/439), EX for 8% (36/439) and T for 6. Outcome for HIV + patients was poorer than for HIV- patients (S: 35/98 versus 291/341, p < 0.00005;PU: 40/98 versus 30/341, p < 0.00005), and the mortality rate of HIV+ patients was higher than that of HIV- patients (EX:18/98 versus 18/341, p < 0.0008). Among those classified PU,no cases of treatment failure were recorded and up to 78% (31/40)were cases of loss of follow-up or abandonment. The variable most closely related to PU was IVDU status (OR = 10.5, p < 0.00005). CONCLUSION: a) Outcomes for the general population are acceptable. b)A marginal group has been identified as characterized by two variables, HIV+ and IVDU, which are the factors associated with poorer outcome of self-administered treatment, indicating that supervised treatment is advisable in cases where those factors are present. PMID- 11412504 TI - [Current and future drug therapies for smoking cessation]. PMID- 11412505 TI - [Skeletal muscle dysfunction in COPD. Cell mechanisms. A.G.N]. PMID- 11412506 TI - [The effects of home visits on compliance with prescription of domiciliary oxygen therapy. A multicenter study]. PMID- 11412507 TI - [Phthisis among Cubans]. PMID- 11412508 TI - [Spontaneous pneumothorax in patients with pulmonary metastasis not undergoing chemotherapy]. PMID- 11412509 TI - [Askin's tumor of the chest wall with early metastasis]. PMID- 11412510 TI - [Tuberculosis in cavum: a case report]. PMID- 11412511 TI - [Pleural empyema and pericardial effusion due to Campylobacter fetus in an immunocompetent adult]. PMID- 11412512 TI - [Several features of nicotine substitution therapy that may lead to clinical problems, based on the information in the technical cards]. PMID- 11412513 TI - [Severity of dyspnea during exercise: similarities and differences between patients with COPD or pulmonary fibrosis]. AB - To understand the mechanisms leading to dyspnea during exercise and to identify possible predictive factors, we compared dyspnea at rest (baseline)and during exercise in 27 patients with chronic obstructive pulmonary disease (COPD) and 39 pulmonary fibrosis (PF) patients. We also compared spirometry and blood gases at rest and after exercise,which consisted of a 12-minute walking test (12 WT). Heart rate and oxygen saturation (SaO2) were recorded every two minutes during the 12 WT. Distance walked was also recorded. Although dyspnea changed during the 12 WT in both groups (p < 0.001),the maximum level of dyspnea reached in the two groups was not statistically different. COPD patients walked farther than did PF patients (782 +/- 182 m vs. 618 +/- 225 m, respectively;p = 0.002) and paused less often during the 12 WT than did PF patients(0.18 +/- 0.55 vs. 0.82 +/- 1.55, respectively; p <0.05). After adjusting for diagnosis, age, sex, baseline dyspnea,distance walked and pauses during the 12 WT, we found that only SaO2 was significantly related to severity of dyspnea during exercise. We conclude that there are important differences in degree of dyspnea experienced during exercise by COPD and PF patients and that SaO2 is the only variable that predicts severity of dyspnea. PMID- 11412514 TI - [Postoperative lung complications and mortality in patients with mild-to-moderate COPD undergoing elective general surgery]. AB - OBJECTIVE: To verify the incidence of postoperative pulmonary complications (PPC) and mortality inpatients with mild-to-moderate chronic obstructive pulmonary disease(COPD) who undergo elective general surgery. Incidence of PPC and mortality were studied in relation to sex, age, anesthesia,surgical incision, duration of surgery, smoking, respiratory symptoms, comorbidity, nutritional status, lung examination, abnormal electrocardiogram,and PaO2, PaCO2, FEV1 and FEV1/FVC. DESIGN: Prospective, open study. MATERIAL AND METHODS: Fifty-nine COPD patients were enrolled (FEV1/FVC< 88% of reference for women and < 89% for men) and studied at a tertiary care university hospital. The patients were examined during the pre-operative period and followed until discharge. RESULTS: Twenty patients (33.9%) experienced PPC and 6 died, two (3.4%) from lung-related causes. Thirty-five PPC events occurred: pneumonia (37.2%), bronchospasm(22.9%), atelectasis (11.4%), acute respiratory insufficiency(11.4%), prolonged mechanical ventilation (11.4%) and bronchial infection (5.7%). Risk factors for PPC were male gender, duration of surgery over 270 minutes, low FEV1/FVC (71.9 +/-10.9%) and surgical incision in the chest or upper abdomen. No significant difference between patients with or without PPC were found for age, presence of respiratory symptoms, comorbidity,abnormal lung examination, nutritional status, smoking, abnormal electrocardiogram, PaO2, PaCO2, FEV1 or duration of pre-operative hospitalization. The rate of PPC was higher in patients smoking more than a mean 40 packs of cigarettes per year. Patients with PPC had longer hospital stays (16.6 +/-15.0 vs. 7.5 +/- 5.7 days) and stayed longer in intensive care units (7.0 +/- 5.9 vs. 1.7 +/- 0.7 days) than did those with no complications (p < 0.05). CONCLUSIONS: The incidence of PPC was 33.9% and lung-related mortality was 3.4%. Risk factors were male gender, amount of smoking, duration of surgery over 270 minutes, low FEV1/FVC, and chest or upper abdominal incision. No risk factor was found to predict mortality in this group. PMID- 11412515 TI - [Thymectomy in myasthenia gravis]. AB - OBJECTIVE: To analyze outcome after thymectomy in patients with myasthenia gravis (MG). MATERIAL AND METHODS: Thirty-five patients with MG underwent surgery in our service between June 1987 and June 1998. Ten had associated thymomas. Preoperative Osserman classification showed 2 at level I, 20 at level IIA, 11 at level IIB and 2 at level III. Extended thymectomy through a medial sternotomy was performed in all. RESULTS: Postoperative complications developed in three patients (1 medullary aplasia, 1 postoperative reintubation, 1 myasthenic crisis). Mean follow-up was 89 months, with 22.8% achieving complete remission and 97.1% reporting improvements. The results were similar in the 10 patients with thymomas (20% full remission and 90% showing improvement). By DeFilippi classification, 22.8% were in class 1, 22.8% in class 2, 51.4% in class 3 and 2.8% in class 4. By Osserman classification, 9 were in the same category before and after surgery, 12 had improved one level, 10 had improved 2 levels, 3 had improved 3 levels and 1 patient had improved 4 levels. CONCLUSION: Thymectomy is an appropriate therapeutic procedure in the multidisciplinary treatment of patients with MG and it is the approach of choice for patients with associated thymomas. The intra- and post-operative complication rate is low and the rate of clinical improvement is high. PMID- 11412516 TI - [COPD and lung cancer: practical implications]. PMID- 11412517 TI - [Risk factors in asthma]. PMID- 11412518 TI - [Systemic factors with potential effect on peripheral muscle function in COPD patients]. PMID- 11412519 TI - [Giant bulla and lung cancer]. AB - Several cases of lung cancer lying within giant bullae have been reported in recent years, such that some authors have posited an association between the two diseases. We describe a patient with a diagnosis of giant bulla in the right lung who presented carcinoma of the lung found within the bulla ten years later. After carrying out a MEDLINE (CD-ROM 1980 to 2000) search of the literature, we agree with other authors on the need to follow the course of patients with bullous lung disease in order to detect the possible development of lung cancer and initiate early treatment. PMID- 11412520 TI - [Recurrent pneumonia and empyema due to Klebsiella pneumoniae as a complication of laparoscopic cholecystectomy]. AB - We report a patient with pneumonia and empyema secondary to an abdominal abscess due to Klebsiella pneumoniae from stones released into the abdominal cavity during laparoscopic cholecystectomy. PMID- 11412521 TI - [Infection due to Streptococcus zooepidemicus in a patient with bronchogenic carcinoma: a rare zoonosis]. PMID- 11412522 TI - [Isolated pleural effusion as unusual manifestation of Chlamydia pneumoniae infection]. PMID- 11412523 TI - [Paradoxical embolism]. PMID- 11412524 TI - [Sputum induction for the diagnosis and monitoring of respiratory disease]. PMID- 11412525 TI - [Effectiveness and efficiency of an outpatient clinic for corticosteroid dependent asthmatics]. AB - OBJECTIVE: To evaluate the efficacy and efficiency of a specialist outpatient clinic for corticosteroid-dependent asthmatics. The clinic was supervised by the respiratory medicine service of a reference hospital. MATERIAL AND METHODS: The first 20 consecutive patients (mean age 58.1+/- 9.5 years; 14 women, 6 men) treated at a specialist outpatient clinic for corticosteroid-dependent asthmatics were studied, with prospective follow-up of 12.33 +/- 4.6 months. The following variables were examined: a) forced spirometry (FS), b)corticosteroid doses, c) number and cost (NC) of visits to the outpatient clinic, d) NC of FS, e) NC of emergency room visits, f) NC of hospitalizations, g)cumulative cost of health care generated by these patients within the National Health Service of Catalonia (NHSC). The results were compared with those recorded in each patient's history. RESULTS: Findings were a) improved FEV1 (55.1 +/- 21.6% vs.60.1 +/- 21%, p = 0.02); b) decreased corticosteroid use (21.9 +/- 11.2 mg vs. 12.8 +/- 6.0 mg, p < 0.0001);c) statistically significant increase in NC to the outpatient clinic and NC of FS; d) statistically significant decrease in number of visits to emergency services and hospitalizations;e) reduced total cost of health care for these patients borne by the NHSC, which went from 4,400,070 Spanish pesetas to 1,171,157 Spanish pesetas. A hospital deficit of nearly 2,000,000 Spanish pesetas was canceled. CONCLUSIONS: Changing the system for delivering health care to these patients has led to improved care (effectiveness) and a noteworthy reduction in cost (efficiency). We conclude that medical specialists should play an important role in reorganizing the present health care system of the NHSC. PMID- 11412526 TI - [Environmental mycobacterial diseases in patients with and without HIV infection: epidemiology and clinical course]. AB - The objective of the present study was to ascertain the clinical features, risk factors, microbiologic spectrum and course of disease after treatment of infections by environmental mycobacteria (EM) in patients with and without HIV infection in our community. Eighty-eight patients with diseases caused by EM diagnosed between 1989 and 1997 were studied; 46 (52.7%) were HIV-positive. Mycobacterium kansasii was the most prevalent pathogen (54%) overall, followed by M. avium complex (40%). However, M. avium complex was most prevalent among HIV positive patients (61%) and M. kansasii was most prevalent among HIV-negative patients (76%). Localized lung infections were most common among HIV-negative patients, whereas 74% of HIV-positive patients had disseminated disease. Among HIV-negative patients, chronic obstructive pulmonary disease and corticosteroid use were common associations. Pulmonary disease was subacute and non-specific in both patient groups, whereas abdominal pain was the first symptom of most patients with disseminated disease. On the chest films of 76% of the HIV-negative patients, we observed cavitation and infiltrates; 60% of HIV-negative patients had normal x-rays. No differences in antibiotic sensitivity were observed between strains from HIV-positive and HIV-negative patients. The prognosis was good in the HIV-negative group with combined therapy with 2 to 4 first-line antituberculous drugs, whereas response was poor in HIV-positive patients in spite of prolonged treatment with 3 to 5 drugs. Nevertheless, thanks to the highly effective anti-retroviral treatment of recent years, we seem to be observing improved response to therapy with less aggressive forms of EM disease. PMID- 11412527 TI - [Model of mortality risk in stage I non-small cell bronchogenic carcinoma]. AB - OBJECTIVE: To develop and validate a mortality risk model for patients with resected stage I non-small cell bronchogenic carcinoma (NSCBC). PATIENTS AND METHOD: Tumors from 798 patients with diagnoses of NSCBC were resected and classified in stage I. The Kaplan-Meier method and Cox's proportional hazard model were used to analyze the influence of clinical and pathologic variables on survival. RESULTS: Univariate analysis revealed that age (p = 0.0461), symptoms (p = 0.0383), histology (p = 0.0489) and tumor size (p = 0.0002) and invasion (p = 0.0010) affected survival. Size (p = 0.0000) and age (p = 0.0269) were entered into multivariate analysis. Each patient's risk was estimated by applying the regression equation derived from multivariate analysis; the mean was 1.47 +/- 0.31 (range 0.68 to 2.92). The series was divided into three groups by degree of risk (low, intermediate and high), establishing the cutoff points at 1.16 and 1.78 (standard deviation of the mean). Five-year survival rates were 85%, 62% and 46%, respectively (p = 0.0000). To validate the model's predictive capacity, the series was divided randomly into two groups: the study group with 403 patients and the validation group with 395. Age (p = 0.0295), symptoms (p = 0.0396), tumor size (p = 0.0010) and invasion (p = 0.0010) affected survival in the univariate analysis. Size (p = 0.0000) and age (p = 0.0358) were entered into Cox's model. Mean risk was 1.94 +/- 0.36 (range 0.98 to 3.32). The series was divided into three risk groups, with cut-off points established at 1.58 and 2.30. Five year survival rates were 90%, 62% and 46% for the low, intermediate and high risk groups, respectively (p = 0.0000). The same model proved able to identify risk when applied to the validation group, in which five-year survival rates were 78%, 61% and 48%, respectively (p = 0.0000). CONCLUSIONS: Risk models can identify patient subgroups, potentially influenced by co-adjuvant treatment, as well as facilitate comparison of patient series. PMID- 11412528 TI - [Cough-inducing capsaicin challenge test in a healthy population]. AB - OBJECTIVE: Cough-inducing agents are used to study the cough reflex in both pharmacological and physiological research. Clinical use of cough challenge testing as a tool in diagnostic algorithms has been limited by the lack of either uniform methodology or reference values for healthy subjects. The objective of this study was to determine the cough threshold for capsaicin in a sample of healthy subjects, while also evaluating the influence of age, sex and smoking. We also assessed the reliability and repeatability of the test. MATERIAL AND METHODS: Ninety-two healthy subjects were enrolled and given a capsaicin challenge test using concentrations from 0.49 to 500mg. The concentrations of capsaicin that triggered two and five coughs, C2 and C5 respectively, were recorded. The test was repeated three days later in a sub-sample of 30 subjects. RESULTS: No subject showed clinical or functional signs of bronchoconstriction. The mean values obtained for the study population were log C2:1.5 +/- 0.69 mM. and log C5: 2.1 +/- 0.4 microM. The geometric means were 31.6 and 134.8 microM for C2 and C5, respectively. No significant differences were found for sex or smoking; however age was significantly related to log C5 (r: -0.27, p < 0.05). Repeatability of the capsaicin test was high, with intra-class correlation coefficients and 95% confidence intervals of 0.75 (0.53-0.87) and 0.88 (0.76 0.97)for C2 and C5, respectively. CONCLUSION: The capsaicin challenge test was safe and easy to administer,gave reproducible results and allowed us to obtain reference values for a healthy population in our community. PMID- 11412529 TI - [Clinical guidelines for the diagnosis and treatment of chronic obstructive pulmonary disease]. PMID- 11412530 TI - [Assessment of peripheral muscle function]. PMID- 11412531 TI - [Recurrent pneumonia secondary to idiopathic bronchomalacia]. AB - Bronchomalacia in adults develops secondary to bronchial disease processes (chronic bronchitis, tuberculosis, neoplasm) or to lesions resulting from prolonged intubation, surgery, trauma, or lung transplantation. We report the case of an 84-year-old woman with no history of any of the aforementioned associations in whom we detected severe idiopathic bronchomalacia of the main left bronchus. The identified lesion caused recurrent pneumonias leading to death. We show images obtained by bronchoscopy, revealing the severity of the lesion and its functional repercussions. PMID- 11412532 TI - [Traumatic asphyxia syndrome]. PMID- 11412533 TI - [Factors predictive of pleural thickening in tuberculous pleuritis]. PMID- 11412534 TI - [Hemorrhagic pleural effusion: atypical presentation of a pleural fibrous tumor]. PMID- 11412537 TI - [Impact of the implementation of the European working standard of 48 weekly hours on primary care]. PMID- 11412538 TI - [Control of smoking from primary care and the Second Week Without Smoking: we have been working for a long time]. PMID- 11412539 TI - [Quality control of the monitoring of normal pregnancy in primary care]. AB - OBJECTIVES: GENERAL: to increase the quality of mother-baby care in our population. SPECIFIC: to improve by at least 10% the record of activities with poorest compliance at the first evaluation on the quality cycle. DESIGN: Quality cycle of the normal pregnancy monitoring programme in primary care (PC). The scientific-technical dimension of quality was the one studied. There was a retrospective, crossed evaluation, on internal initiative, about the clinical records of the pregnant women. SEQUENCE: first evaluation of pregnancies in 1994 95; application of corrective measures in 1996; second evaluation of pregnancies in 1997-98. SETTING: Primary care. PATIENTS: All the pregnancies monitored by our PC team during the periods described. MEASUREMENTS AND MAIN RESULTS: CRITERIA EVALUATED: care activities of the programme for monitoring normal pregnancies in Area 10. First evaluation (n = 54): 11.5% overall non-compliance. The criteria with under 80% compliance were: folic acid supplement, counselling, diagnosis of pregnancy diabetes and attendance in the month after delivery. Second evaluation (n = 69): 6.3% overall non-compliance. All activities surpassed 80% compliance except obstetric examination on the first visit, attendance a week after delivery and attendance a month after delivery. CONCLUSIONS: After introducing certain simple corrective measures, we achieved overall improvement of over 40% and increased compliance in most activities, except for attendance after delivery and obstetric examination on the first visit. We think a continuous cycle of quality improvement needs to be maintained in order to achieve adequate mother-baby care. PMID- 11412540 TI - [Variation over two years of cardiovascular risk in patients cared for at primary care services for hypertension, diabetes and dyslipidemia]. AB - OBJECTIVE: To analyse the modifications in cardiovascular risk in patients seen over two years in adult services for dyslipaemia, diabetes mellitus or hypertension. DESIGN: Observational, retrospective study with repeated measurement tests. SETTING: Two health centres. PATIENTS: 265 patients attended by the above services who were free of cardiovascular disease at the start of the study. MEASUREMENTS AND RESULTS: During the study period, the cumulative incidence of cardiovascular morbidity was 4.2% (1.8-6.6%). Patients, without events, underwent a mean reduction in overall cardiovascular risk of -1.6% (p < 0.001). In patients classified as high-risk at the start of the study, the mean reduction was much greater: -3.7% (p < 0.001). Giving up smoking and controlling dyslipaemia brought about the greatest reductions in cardiovascular risk. CONCLUSIONS: Monitoring of cardiovascular risk in patients attended for conditions in the INSALUD portfolio of services, i.e. hypertension, dyslipaemia, diabetes mellitus, enables this risk to be reduced and priority to be given to health care delivery to more vulnerable groups. PMID- 11412541 TI - [Acceptance of lot sampling: its applicability to the evaluation of the primary care services portfolio]. AB - OBJECTIVE: To determine the applicability of the acceptance of lot quality assurance sampling (LQAS) in the primary care service portfolio, comparing its results with those given by classic evaluation. DESIGN: Compliance with the minimum technical norms (MTN) of the service of diabetic care was evaluated through the classic methodology (confidence 95%, accuracy 5%, representativeness of area, sample of 376 histories) and by LQAS (confidence 95%, power 80%, representativeness of primary care team (PCT), defining a lot by MTN and PCT, sample of 13 histories/PCT). Effort, information obtained and its operative nature were assessed. SETTING: 44 PCTs from Murcia Primary Care Region. MEASUREMENTS AND MAIN RESULTS: Classic methodology: compliance with MTN ranged between 91.1% (diagnosis, 95% CI, 84.2-94.0) and 30% (repercussion in viscera, 95% CI, 25.4-34.6). Objectives in three MTN were reached (diagnosis, history and EKG). LQAS: no MTN was accepted in all the PCTs, <<01-diagnosis>> being the most accepted (42 PCT, 95.6%) and <<07-Funduscopy>> the least accepted (24 PCT, 55.6%). In 9 PCT all were accepted (20.4%), and in 2 none were accepted (4.5%). Data were analysed through Pareto charts. CONCLUSIONS: Classic methodology offered accurate results, but did not identify which centres were those that did not comply (general focus). LQAS was preferable for evaluating MTN and probably coverage because: 1) it uses small samples, which foment internal quality improvement initiatives; 2) it is easy and rapid to execute; 3) it identifies the PCT and criteria where there is an opportunity for improvement (specific focus), and 4) it can be used operatively for monitoring. PMID- 11412542 TI - [Multicenter study of children's compliance to antibiotic treatment in primary care]. AB - OBJECTIVES: To evaluate the degree of children's adherence to prescription of antibiotics and to determine factors linked to this adherence. DESIGN: Cross sectional study, conducted through a telephone survey of parents of the children under study ten to fifteen days after the prescription of the antibiotic. SETTING: Thirty primary care centres in 6 Spanish provinces. PATIENTS AND OTHER PARTICIPANTS: Parents or guardians of children from 0 to 10 who attended the primary care centres between October 1998 and January 1999 for treatment of an acute infection and who were prescribed an oral antibiotic treatment with two or more daily doses. Interventions. The measuring instrument was the Morisky-Green test. MEASUREMENTS AND MAIN RESULTS: 2244 cases were studied. 1043 of them complied adequately (46.5%; 95% CI, 44.4-48.5). The factors associated with adherence were children's school situation, the age of parents or carers and the number of daily doses. CONCLUSIONS: Only half the children complied correctly with the treatment indicated. Pre-school children, those with parents or carers over 40 and those with a prescription of under three daily doses followed the treatment better. These factors need to be borne in mind by paediatricians when they prescribe an antibiotic. PMID- 11412543 TI - [Treatment compliance: what do we know about Spain?]. PMID- 11412544 TI - [Cost-benefit analysis]. PMID- 11412545 TI - [Pediatrics and Internet]. PMID- 11412546 TI - [A debate should never be an exchange of subjective opinions]. PMID- 11412547 TI - [Fibromyalgia]. PMID- 11412548 TI - [Informatization in primary care]. PMID- 11412549 TI - [Inappropriate use of third generation cephalosporins in primary care]. PMID- 11412550 TI - [Form for chronic patients: improving registries, evaluations, and health education]. PMID- 11412552 TI - [Ethical dilemmas in caring for terminal patients: a view from primary care]. PMID- 11412553 TI - [Prevalence and characteristics of major depression and dysthymia in primary care]. AB - OBJECTIVE: To find the prevalence and characteristics of depressive disorders in patients consulting in primary care.Design. Cross-sectional descriptive study. Two stages: screening, then structured psychiatric interview.Setting. Eight clinics at four primary care centres. PATIENTS AND METHODS: A sample of 350 consecutive patients aged between 18 and 70 filled out Zung's Self-Rating Depression Scale (SDS). The diagnoses of major depressive episode and dysthymia for the 138 with positive result and the 67 with negative result were investigated through the Structured Clinical Interview for DSM-IV Axis I Disorders. Measurements and main results. We found a weighted prevalence of 14.7% (95% CI: 10.7-18.7) for major depression and 4.6% (95% CI: 2.4-6.8) for dysthymia. Mean score on the SDS was 65.6 (SD 11.6) in the group with major depression and 63.3 (SD 8.7) in the group with dysthymia. Mean score in the not depressed group was 44.2 (SD 8.7), lower than the scores for both groups with depression (p < 0.0001). Being female distinguished the depressed groups from the not-depressed group. Lower educational level and the amount of over-users distinguished the group with major depression from the not-depressed group. The symptom profiles were virtually identical for the two depressed groups, whereas the frequency of occurrence of most of the symptoms explored differed significantly between those with and without depression. CONCLUSIONS: Prevalence of major depression and dysthymia are high in primary care patients in our area. Both disorders have common characteristics of demographic variables and symptom profile. PMID- 11412554 TI - [Tobacco detoxication at a primary care clinic: efficacy of medical counseling, minimal intervention and nicotine replacement therapy at the one-year follow-up]. AB - OBJECTIVE: The main objective was to compare the efficacy of one-off tobacco counselling and of the systematic minimum intervention on making people give up smoking. A secondary aim was to evaluate the efficacy of nicotine replacement therapy (NRT) in the context of a primary care consultation. DESIGN: Controlled and randomised clinical trial. SETTING: Primary care. PARTICIPANTS: Smokers who over 12 months attended a primary care clinic at an urban health centre (304 patients). INTERVENTIONS: Two kinds of intervention were conducted at random on patients with low nicotine dependence (one-off medical counselling and counselling integrated into what is known as the minimal intervention, but conducted systematically). NRT was administered through nicotine patches (11% nicotine) to patients with moderate-to-high nicotine dependence. MEASUREMENTS AND MAIN RESULTS: Short-term cessation a year after the minimal intervention was 39% (29.4-49.3%), and maintained cessation 30.9% (29.4-49.3%), as against 11% (5.6 18.8%) short-term cessation in the group that received one-off counselling (p < 0.0001). Short-term tobacco cessation in the NRT group was 35.3% (24.1-47.8%), and maintained cessation 30.8%. CONCLUSIONS: Primary care is a suitable context for an intervention against tobacco dependency, through the use of any of the established interventions: one-off medical counselling, systematic minimal intervention, or specialist drug treatment through NRT. Therefore, these kinds of intervention must form part of PC clinics' daily activity. PMID- 11412555 TI - [Tuberculosis trends in Palencia]. AB - OBJECTIVES: To describe the most important epidemiological characteristics and the tendency of the incidence of tuberculosis in Palencia from 1986 to 1999. DESIGN: Descriptive study. SETTING: Palencia province. PATIENTS: 1158 cases of pulmonary tuberculosis and 177 cases of extrapulmonary tuberculosis. MEASUREMENTS AND RESULTS: We reviewed the Obligatory Diseases Declaration and their epidemiological characteristics. We used descriptive statistics, 2 and Student tests, sensibility and positive predictive tests. There were 1158 cases of pulmonary tuberculosis and 177 of other tuberculosis. Tuberculosis was more frequent in men (69.2%). The most affected groups of age were 20-29 years old (20.6%) and 20-49 years old (48.2%). The group of age between 60 and 79 years old reported an incidence of 21.8%. 33 of 100 notified cases of AIDS had extrapulmonary tuberculosis and 18% had pulmonary tuberculosis. CONCLUSIONS: Tuberculosis is an important problem of public health. It's more common in young men and its epidemiological characteristics are similar to AIDS sickness in our environment. PMID- 11412556 TI - [Suitability of antibiotic prescription in primary care in the Basque Autonomous Community]. AB - OBJECTIVE: To study the variability and suitability of antibiotic prescription in primary care in the Basque Country. DESIGN: Prospective, descriptive study. SETTING: Basque Country health care centres both within and not in the reformed network. MEASUREMENTS AND MAIN RESULTS: 3182 infections from consultations for infectious disease were studied through sampling stratified by health area during February, March and April 1998. The appropriateness of antibiotics (ATB) prescription was compared with some standards after an extensive bibliographic review. 68.6% of consultations were infections not needing ATB (infections of respiratory airways, acute bronchitis, flu and gastro-enteritis). ATB were used in 28.5% of these cases (CI: 26.6-30.5), especially in acute bronchitis (90.7%) (CI: 87.3-93.4). Unnecessary prescription supposes 39.7% of total antibiotic prescription (CI: 37.2-42.1%), which means unnecessary annual expenditure of 1155 million pesetas. In 31.4% of the infections that require ATB treatment, in 23.7% inappropriate treatment was chosen (CI: 21-26.7%). Overall appropriateness of treatment was 72.2% (CI: 70.6-73.8). Working in the reformed system was linked to better prescription in all cases. Paediatricians prescribed better in the case of infections not susceptible to ATB treatment. CONCLUSIONS: ATB prescription is not consistent with the available scientific evidence. Incorrect treatments occur especially in benign and self-limiting processes. A local policy of ATB use that includes multi-disciplinary collaboration and effective ongoing training is necessary. PMID- 11412557 TI - [Do we gain or lose information with computerisation?]. AB - OBJECTIVE: To calculate the concordance of the computer record and the clinical history (CH) in preventive actions and health problems. DESIGN: Cross-sectional descriptive study. Quality evaluation. SETTING: Urban health centre with 31000 inhabitants. PATIENTS AND OTHER PARTICIPANTS: Randomised batch sample, with 14 cases for each of the 8 attendance base units with computerised records since 1997. N = 112. EXCLUSION CRITERIA: no visit later than January 1997 and absence of records in the CH or computer. MEASUREMENTS AND MAIN RESULTS: Through the checking of the records in the CH and computer, a mean concordance of 73.5 (95% CI, 66.8-80.2) for preventive actions and 93.5 for health problems (95% CI, 90.6 96.4) was found. There was a mean computer under-recording for health problems of 6.5% (95% CI, 3.62-9.32), and for preventive actions of 21% (95% CI, 9.1-33.3) only in those actions based on manual activity. However, in preventive actions based on verbal activity there was 14.3% mean CH under-recording (95% CI, 1.15 27.5). CONCLUSIONS: Concordance is not uniform, with under-recording for some parameters detected. This may affect the reliability and validity of health information in these records. We believe that the way data are collected determines this to a large extent. We suggest as corrective measures improving the training and incentives of health professionals, making computer programmes more appropriate to their purpose and standardising data collection in primary care CR. PMID- 11412558 TI - [Statistics and causes of mortality in type-2 diabetes]. AB - OBJECTIVE: To establish the causes of mortality in people with type-2 diabetes and check the reliability of official records. DESIGN: Cross-sectional, descriptive study. SETTING: Urban health centre with a population which has aged and is depressed socially and economically. PATIENTS: Randomised sample of 120 deceased patients with type-2 diabetes, extracted from the total number of deceased patients with type-2 diabetes on the centre's lists over the last five years. INTERVENTIONS: Personal details, clinical data and clinical cause of death were gathered from the history. Then the clinical cause of death recorded in the Civil Registry was obtained. MEASUREMENTS AND MAIN RESULTS: The main cause of death is neoplasm (25.8%), but if we group the cardiovascular causes they add up to 47.4% of the total. According to official statistics, cardiovascular disease only occasioned death in 35% of cases. There was 61.7% overall concordance between the clinical cause and the official cause in the Civil Registry. There was close concordance only in the case of neoplasms (kappa = 0.84). CONCLUSIONS: We found in our study that the main cause of death in type-2 diabetics was cardiovascular disease (47.7%), while in the Civil Registry mortality for this reason is clearly lower, which leads us to question the validity of the official statistics. This suggests we should insist even more on the importance of monitoring cardiovascular risk factors. PMID- 11412559 TI - [Health problems and factors determining the number of on-demand visits of over users of a health centre]. AB - OBJECTIVES: To describe the social and demographic characteristics and health problems of over-users of a health centre and to determine the number of attendances requested on demand at the general medical clinic. DESIGN: Retrospective and descriptive. Multivariate analysis: multiple linear regression. SETTING: Urban health centre. PATIENTS: Patients who requested a consultation at the health centre on at least nine occasions in 1999 (N = 7852). Random sample of 386 patients (alpha 0.05, 95% accuracy). MEASUREMENTS AND MAIN RESULTS: Dependent variable: number of on-demand consultations requested in 1999. INDEPENDENT VARIABLES: age, family size, area of residence, active or pensioner, inclusion in programmes, health problems (CIAP-2), number of drugs on repeat prescription (Anatomical Classification), short-term time off work. 57% of over-users were women (CI, 52.9-62.7%); mean age 55 (SD 18.5); family size 2.7 members (SD 1.457); 58.8% pensioners. Mean number of appointments 15 (SD 6.7). Mean habitual consumption of 1.58 medicines (SD 2.46). Inclusion in programmes: 37.7% hypertension, 16% diabetes, 17% dyslipaemia, 16% nursing clinic. Most prevalent pathologies: cardio-circulatory (43.8%), endocrino-metabolic (32%), traumatology (26.7%) and mental health (21%). The regression model included the variables of age, number of medicines and having had short-term time off. CONCLUSIONS: Over users of our health centre are middle-aged women with chronic physical health problems and with mental health problems. The number of attendances is related to age and administrative requirements: medication and sick notes. PMID- 11412560 TI - [Pharmacy attention to prescriptions in order to bring practice into line with scientific evidence]. AB - OBJECTIVE: The objective of this study is to assess the usefulness of a programme of attention to pharmaceutical prescriptions (APP) so as to detect medicine related problems. DESIGN: A descriptive study of the findings after six months operation of this programme. SETTING: Five primary care centres in the Bahia Cadiz Area. PATIENTS: All the patients (499) who requested advice on their drug therapy and were included at the request and on the criterion of their doctor were studied. INTERVENTION: We introduced the APP programme for chronic patients, with two basic aims: Care: to detect medicine-related problems and propose individual solutions. Teaching: to bring drug-therapy skills used in clinical practice into line with the scientific evidence available. MEASUREMENTS AND RESULTS: We focused on the detection of problems related to the most common medicines, with direct and relevant incidence on morbidity and mortality. We detected 236 cases (47%) of possible therapeutic improvement that had important implications for associated morbidity, of which 114 (23%) also had implications for mortality. 56% of the patients took more than 4 medicines, multiple medication that could easily be reduced in 43.5% of cases, so avoiding the use of medicines of low intrinsic value. CONCLUSIONS: The programme was very useful, as numerous very clinically relevant problems were detected and information was gained that the doctor can use to bring drug therapy into line with the available scientific evidence. PMID- 11412561 TI - [Program of mental health training for family and community medicine residents: the Zaragoza model]. PMID- 11412562 TI - [Establishing priorities]. PMID- 11412563 TI - [Malignant neuroleptic syndrome linked to haloperidol]. PMID- 11412564 TI - [Coordination between primary and specialized care]. PMID- 11412565 TI - [Peripheral arteriopathy: use of the Doppler technique in primary care]. PMID- 11412566 TI - [Post-coital contraception]. PMID- 11412567 TI - [Post-coital contraception]. PMID- 11412568 TI - [Security and confidentiality in computer use in the clinic: do we know the law?]. PMID- 11412570 TI - [Making consultations flexible at demand peaks]. PMID- 11412571 TI - [Can or should blood pressure be measured at pharmacy offices?]. AB - OBJECTIVE: To evaluate concordance between blood-pressure (BP) measurements at the pharmacy office (PhO) and the nurse office (NO) in the health care centre (HCC). DESIGN: Descriptive study. SETTING: Community. METHODS: 36 PhO have voluntarily participated in the province of Albacete, where they have done 3 BP measures, without previous instructions and with their usual measurements devices, for 3-5 subjects, who were referred to their HCC so that they were taken another 3 BP measures in the NO with their Hg sphymomanometer and in their usual measure conditions (blind measures in relation to those taken at the PhO). These subjects were given a stamped envelope to send the BP measurements (taken at the NO), to the Official Pharmacy College. RESULTS: The 6 BP measurements have been completed to 96 subjects with an average of 57.3 years old (women 63%). The differences between PhO and NO were > 5 mmHg in 58 subjects (60.4%) in the case of SBP and in 45 subjects (46.9%) in the case of DBP, and it was more than 15 mmHg in the 17 subjects (17.7%) with SBP and in 9 subjects (9.4%) with DBP. The difference average was 9.5 mmHg (SD, 8.4 mmHg) and 6.4 mmHg (SD, 5.3 mmHg) respectively. The use rate of digit 0 was 22% at the PhO and 46.5% at the NO. In most of PhO, measurement electronic devices have been used for the BP, but not validated for clinical use. CONCLUSIONS: The PhO can be a good place for the hypertension screening, but the chemist must be trained in the BP correct measurement and use validated electronic devices. Standardization measurement conditions and the use of validated electronic devices must be extended to the NO. PMID- 11412572 TI - [Mumps in a urban area of the Community of Madrid. Vaccination status, diagnosis and intervention measures]. AB - OBJECTIVES: To describe the epidemiological situation of an epidemic of mumps, to calculate the risk of catching the illness depending on the vaccine strain used and to evaluate the diagnostic performance of the serologic determination. DESIGN: Longitudinal observational study. The incidence of mumps between January 1999 and June 2000 was calculated. Case-control study of outbreaks evaluated the risk of catching the disease depending on the vaccine strain. SETTING: Health Area 4 in the Community of Madrid. PARTICIPANTS: Residents and students from schools in the district.Interventions. Epidemiological vigilance. Serologic determination of IgM and titration of IgG. In outbreaks, people exposed were vaccinated after assessment of their vaccine status. MEASUREMENTS AND MAIN RESULTS: The rate of incidence in the first 27 weeks of the year 2000 for the 1-4 year-old age group was 334.7/105, against 12.2 in 1999. In children under 5 vaccinated with the Rubini strain there was a significantly greater risk of suffering illness than in those vaccinated with the Jeryl-Lynn strain (OR, 6.28, 95% CI, 1.43-38.50). The amount of cases confirmed serologically (specific IgM positive) was 22.22%. The mean of the logarithms of the IgG titres was significantly higher (p < 0.001) in IgM-negative patients with epidemiological confirmation (4.05, 95% CI, 3.18-4.28) than in IgM-negative cases without any epidemiological link (2.58; 95% CI, 2.33-2.84). CONCLUSIONS: We emphasise the increase of mumps incidence in the school population (fundamentally in the under 5s), which parallels an increase in outbreaks. The results suggest that the Rubini strain is less efficacious. The possibility of using alternative methods to IgM detection (IgG titration) as a diagnostic tool in a population with high vaccination coverage is posed. PMID- 11412573 TI - [Do the new criteria of the American Diabetes Association lead to earlier diagnosis of type-2 diabetes mellitus?]. AB - OBJECTIVE: To analyze the period of time between the first occasional fasting hyperglycaemia (OFH) and the diagnosis of type 2 diabetes mellitus (DM2), using the World Health Organization (WHO) criteria or the American Diabetes Association (ADA) criteria. DESIGN: Retrospective, observational study. SETTING: Urban primary care centre. SUBJECTS: 104 patients with DM2 diagnosed between 1991 and 1995 who had a previous OFH. MEASUREMENTS: Age, gender and other risk factors, dates of the first OFH (fasting plasma glucose >= 110 mg/dl), the diagnosis according to WHO criteria (2 fasting plasma glucose >= 140 mg/dl or >= 200 mg/dl two hours after the oral glucose test tolerance (OGTT)) or with the ADA criteria (2 fasting plasma glucose >= 126 mg/dl), and the intervals in months between them. RESULTS: Of the 222 diagnosed patients, 104 (47%) had previous OFH. Age at diagnosis was 60.8 (SD 10.1) and 53% were women. OGTT was performed in 51 cases (49%). The median (range) of the interval between the first OFH and diagnosis was 16 months (0-101) for those who were undertaken an OGTT, and 45 months (1-104) for those who were not (p = 0.003). In these last ones, ADA criteria reduced the interval to 31 months (0-97) (p < 0.001). In 27 of these patients who did not satisfy both criteria at the same time, ADA criteria reduced the interval to 10 months (0-93) (p < 0.001). CONCLUSIONS: Not performing the OGTT means a delay in diagnosis which can be countered by applying the ADA criteria. PMID- 11412574 TI - [Glaucoma and ocular hypertension in primary care]. AB - OBJECTIVES: To find the distribution of intra-ocular pressure (IOP) in our population over 40 and the prevalence of glaucoma (G) and ocular hypertension (OH). DESIGN: Descriptive. SETTING: Primary care. SUBJECTS: 990 people >= 40 years old.Measurements. Age, sex, body mass index (BMI), ophthalmic history, risk factors, visual sharpness (optotype), ophthalmoscopy and IOP (applanation tonometry). Patients with back-of-eye disorders and/or IOP >= 21 mmHg were referred to the ophthalmologist for assessment (campimeter). RESULTS: 870 people, average age 54.66 and 45.4% of them men. Mean IOP was 12.96 in the right eye and 13.27 in the left, with significant differences between them, but not between sexes or age-groups. 70 patients were referred (11 for IOP >= 21, 54 for disturbance perceived by ophthalmoscope and 5 for both reasons). The ophthalmologist classified 9 as G, 12 as OH and 15 as suspected G. We calculated 1.81% (95% CI, 0.98-2.63) prevalence of G in the >= 40s, including 9 cases already known. OH prevalence was 1.61% (95% CI, 0.82-2.39). Through logistical regression, we found greater risk of G in people with myopia (OR adjusted for age and sex = 3.01) and Hypertriglyceridaemia (ORa, 6.34). OH risk was significantly greater in patients with BMI >= 30 (ORa, >= 4.20). CONCLUSIONS: Glaucoma prevalence confirms published findings from other similar populations, while IOP and OH prevalence were much less here. Given that half G cases are undiagnosed, we believe its early detection in primary care should be highlighted, at least in at-risk groups. PMID- 11412575 TI - [Cardiovascular risk and the new diagnostic categories for diabetes mellitus proposed by the American Diabetes Association]. AB - OBJECTIVE: To study the cardiovascular risk (CVR) associated with the diagnostic criteria for diabetes proposed by the American Diabetes Association. DESIGN: Cross-sectional, descriptive study. SETTING: Urban health district. PATIENTS: 1840 patients > 14 years old, selected by simple randomised sampling from the clinical records (CR) archive, were studied. METHOD: Through review of the CR, the patients were classified as having: normal glycaemia, disturbed basal glycaemia (DBG) and type-2 diabetes mellitus (DM2). CVR was studied through the simplified Framingham method and the CVR factors of obesity, tobacco dependency, hypertension, hypercholesterolaemia and hypertriglyceridaemia. The likelihood of having high or very high CVR and the CVR factors described in the patients with DBG or DM2 were compared with the same in those with normal glycaemia, through logistical regression with the odds ratio adjusted for age and sex. RESULTS: 1351 patients were classified: 995 with normal glycaemia, 146 with DBG and 210 with DM2. Patients with DBG or DM2 had greater likelihood of high or very high CVR, with some OR at 2.26 (95% CI, 1.39-3.69) and 2.74 (95% CI, 1.81-4.15), respectively. They also had differences (p < 0.05) for obesity (OR, 1.76 and 1.58), hypertension (OR, 1.75 and 2.15) and hypertriglyceridaemia (OR, 1.73 and 2.70), respectively. There were no differences (p > 0.05) for tobacco dependency and hypercholesterolaemia. No differences were found (p > 0.05) between DBG and DM2 for CVR and the CVR factors studied. CONCLUSIONS: Patients with DBG and DM2 are at high CVR. PMID- 11412576 TI - [Tackling mental health problems from primary care: relationship with specialized support services]. AB - OBJECTIVES: To find the relationship between primary care (PC) and mental health (MH) by evaluating: 1) perception of MH problems in PC: training of professionals and doctor-patient relationship, and 2) relationship between PC and specialist support services: coordination meetings and reasons for referrals. DESIGN: Descriptive study. SETTING: Seven PC centres that share the same mental health centre in the Girona Health Region. PARTICIPANTS: All the PC doctors from the 7 centres (n = 66). MEASUREMENTS AND MAIN RESULTS: Specific, anonymous and self administered survey, with 71 Likert-type items, distributed in 14 fields. 51 replies (77.2%). 60.4% valued positively their MH training. 100% thought that a good doctor-patient relationship improves adherence to treatment, and 76.5% saw this hindered by lack of time. Aggressive and manipulative patients caused a higher level of tension (70.6% and 66.7%). The point most highly valued of their relationship with the MH centre was its support in patient follow-up (69.6%) and the agreement to share responsibilities for treatment of mental disorders (71.1%). The forms of support most valued were: inter-clinical meetings (80.4%) and inter-clinic telephone consultations (78.4%). The factors most conditioning referral to the MH centre were ideas of suicide (96.1%) and the seriousness of the symptoms (94%). CONCLUSIONS: Greater MH training should be offered to PC doctors. PC doctors think they have little time available to devote to their psychiatric patients. Availability of a specialist PC support team is essential. PMID- 11412577 TI - [Risk factors of a new cardio-embolic cerebral accident in non-valvular atrial fibrillation treated with acenocoumarol]. AB - OBJECTIVES: To investigate the risk factors determining the appearance of a new cardio-embolic cerebral accident on secondary thrombophylaxis of non-valvular auricular fibrillation (NVAF). DESIGN: Controlled observational study on a series of patients treated with acenocoumarol for a period of 2.8 years. SETTING: Anticoagulation Unit of the La Paz Hospital and patients from Madrid Area 5. PARTICIPANTS: 172 patients with NVAF, over 55 and with fibrillation for at least a year, and who had suffered at least one ischaemic cerebral accident. All were treated with acenocoumarol and controlled at an INR of 2.5. The risk factors were determined and the appearance of new cardio-embolic phenomena was watched during the study period. MEASUREMENTS AND MAIN RESULTS: 12 cardio-embolic phenomena were recorded (11 CVA and one peripheral embolism). The univariate study showed there was a significant association between patients who suffered a new ictus and those who suffered heart failure (p < 0.05) and had a history of more than one ictus before the start of the study (p < 0.05). Multivariate analysis found that what had independent predictive value was having suffered more than one ictus. CONCLUSIONS: The greatest threat for developing a new ischaemic CVA during anticoagulation in NVAF is having a history of two or more previous ictus, as if the tendency to recur was due to these patients having more extensive atherosclerosis. PMID- 11412578 TI - [Tobacco in Barcelona's metropolitan transportation: observation study]. AB - OBJECTIVE: To describe the extent of no-smoking notices, consumption and sales of tobacco in Barcelona's underground transport installations. DESIGN: Descriptive, observation study. SETTING: Community. METHODS: A sample of 54 stations was selected at random. The presence of users smoking, staff smoking, notices about tobacco consumption and tobacco sales were evaluated. INTERACTIONS: Platforms, entrance hall and corridors and/or stairs were observed for at least 5 minutes at each station. MEASUREMENTS AND MAIN RESULTS: Users smoking were recorded in 87% of the observations, and cigarette stubs in 97.5% of observations, whereas company staff were only seen smoking in 5.6%. In 72.2% of the entrances there were tobacco sales-points. The users observed smoking were divided equally between the sexes, although the men were mainly middle-aged adults (64.3%) and the women were mainly young (60.6%). In 64.2% of the observations, some sort of notice was seen. CONCLUSIONS: Both notices and compliance by staff are high, although users habitually consume tobacco. Tobacco sales-points at stations are very common. The findings justify the adoption of specific measures designed to improve compliance with the regulations. PMID- 11412579 TI - [Gastrointestinal signs and symptoms during pregnancy and postpartum in a sample of Spanish women]. AB - OBJECTIVE: To describe the evolution of the signs and more frequent gastrointestinal symptoms during the pregnancy and in the two weeks later the birth. METHODS: SUBJECTS: 583 pregnant women, elects in an accidental way, interviewees between the 8 and 12 weeks of gestation (n = 133), among 18 and 22 (n = 155), among 37 and 42 (n = 203) and two weeks after the childbirth (n = 92) and coming from centers of health and hospitals of the Comunidad Valenciana and Murcia. The questionnaire, answered in a voluntary way by interview, it contained a listing on the signs and more frequent gastrointestinal symptoms during the pregnancy, and she answered on the presence or not of the same ones in the two weeks previous to the moment of the interview. RESULTS: The percentage of nauseas and vomits, although higher in the first trimester, stays around the 25-30% in the third gestation trimester. The heartburn is presented in 58.6% of the pregnant women in the third trimester. The sialorrea is presented in 7.7% of all the pregnant women and she stays during the whole gestation. The constipation stays constant during the gestation above at the 25-30%. The prevalence of haemorrhoids to the beginning of the gestation is over to 8% to be located in the postpartum in 53.3%. CONCLUSIONS: The description of the evolution of the signs and symptoms of the pregnancy described in the obstetrics manuals don't come up with our data. PMID- 11412580 TI - [Emergency contraception]. PMID- 11412581 TI - [Clinical pathways in primary care]. PMID- 11412582 TI - [Clinical practice guidelines on the Internet]. PMID- 11412583 TI - [Quality of minor surgery in primary care. Are current indicators adequate?]. PMID- 11412584 TI - [Ischaemic cardiopathy: do we practice secondary prevention in primary care?]. PMID- 11412585 TI - [Vasculitis linked to biotin use]. PMID- 11412586 TI - [Prevalence and associated risk factors of smoking among high school students in the city of Lleida]. PMID- 11412587 TI - [PFAPA syndrome: periodic fever, aphthous stomatitis, pharyngitis and adenitis]. PMID- 11412589 TI - [Evaluation of the utility and efficacy of a day hospital in a gastroenterology department]. AB - Because public health funds are limited, full advantage should be taken of the resources available. Consequently, hospital stay should be reduced with no loss of health care quality. In our Gastroenterology Department, investigations or treatment requiring a short hospital stay are carried out in a day hospital. Between March 1998 and March 2000, 2,169 patients were treated: 1,862 outpatients and 307 patients referred from another hospital. A total of 95.2% of the outpatients were discharged. Hospital stay was prolonged in 4.8% (91/1,862): 13 due to incomplete investigations, 29 because of worsening of their underlying disease before the procedure and 49 due to complications. The overall complication rate was 2.8% (62/2,169). Six complications developed after discharge; of these two appeared within 24 hours. After the opening of the day hospital, the mean hospital stay in the Gastroenterology Department was reduced from 9.05 days to 6.07 days (p = 0.001). In conclusion, the use of a day hospital in a gastroenterology department is useful, effective, safe and profitable. PMID- 11412590 TI - [Epidemiology of chronic inflammatory bowel disease in Gijon, Asturias]. AB - AIMS: To determine several aspects of the epidemiology of chronic inflammatory bowel disease (IBD), including distribution of the various forms of IBD, sex, age at diagnosis considering lag-time to diagnosis, criteria used in the diagnosis, the relationship between educational level and activity, familial aggregation, phenotype (site and clinical type), number of admissions and mean hospital stay/year, surgical requirements and mortality. PATIENTS AND METHODS: We carried out a retrospective (1954-1993) and prospective (1994-97) descriptive epidemiologic population study, in the fifth health district of Gijon in Asturias (Spain), with 225,798 inhabitants. A total of 595 patients diagnosed with chronic IBD according to the diagnostic criteria described by Lennard-Jones and Truelove for Crohn's disease (CD) and ulcerative colitis (UC) and according to Ashley B. Price's criteria for indeterminate colitis (IC) were studied. In all patients a complete clinical follow-up was performed. RESULTS: During the study period, we diagnosed 595 patients with IBD [305 patients with UC (51.3%), 272 (45.7%) patients with CD and 18 (3%) with IC]. Sex distribution was 287 females (48.2%) and 308 males (51.8%), with a male/female ratio of 1.07. Mean age at presentation was 38.79 +/- 17.44 years (UC: 43.37 +/- 17.55; CD: 33.98 +/- 16.16; IC: 33.73 +/ 13.48), p = 0.000. Age at onset prior to diagnosis was as follows: UC: 42.03; CD: 30.47; IC: 30.99 (p = 0.000). Diagnostic criteria used in UC was symptomatic in 99.01% (p = ns), endoscopic in 95.04% (p = 0.000), and pathologic in 87.21% (p = 0.000); in CD diagnostic criteria used was radiologic in 85.29% (p = 0.000). A total of 29.1% of patients with UC and 66.7% of those with CD had higher education (p = 0.0005). Family history was found in 9.8%. Anatomical site was as follows: in UC: rectum 21%, 28.2% rectum and sigmoid, 22.3% left colitis, 4.2% distal to hepatic flexure and 24% pancolitis; in CD: 32.72% terminal ileum, 19.11% colon, 37.13% ileo-colon, 11.02% extensive intestinal and 3.67% gastro duodenal. A total of 8.37% of patients with UC and 14.51% of those with CD had been hospitalized during the previous 4 years; mean hospital stay was 1.63 days in UC and 2.27 days in CD. The mean surgical requirements were 0.54 +/- 1.08 (31.59%); UC: 0.11 +/- 0.36 (10.2%); CD: 1.04 +/- 1.38 (56.25%), p = 0.000. The mortality rate was 48.73 deaths/1,000 inhabitants (UC: T = 65.57; CD: T = 33.08; IC:T= 0) p = ns. The standardized mortality ratio was 4.83 (UC: 6.51; CD: 3.28). CONCLUSIONS: We highlight the uniformity of the distribution of IBD in relation to types of disease and sex. Patients with CD had a higher level of education. Genetic components play an important role in these diseases and familial aggregation was high, especially in CD. Complicated situations are infrequent in this group of patients. Morbidity was higher in patients with CD as reflected by surgical requirements and hospital stay. Mortality was lower in CD than in UC. PMID- 11412591 TI - [Adult celiac disease: study of 21 cases and review of the bibliography]. AB - Celiac sprue is a chronic disease characterized by a typical lesion in the small intestine and nutrient malabsorption that improves on withdrawal of dietary gluten. Adult celiac disease (or gluten enteropathy) is a common and under diagnosed disease since presentation varies among a) symptomatic or clinically active forms (including mono- or oligosymptomatic forms), b) asymptomatic, subclinical or silent forms, and c) latent forms. We present a descriptive study of a retrospective series of patients diagnosed with symptomatic or clinically active adult celiac disease who required hospitalization between January 1993 and June 2000. The form of symptomatic expression, biological alterations, investigations that confirm the diagnosis, associated diseases, complications and response to treatment are described. PMID- 11412592 TI - [Acute hepatitis due to ingestion of Teucrium chamaedrys infusions]. AB - Herbal remedies, used for many years for therapeutic purposes, have traditionally been considered safe and effective. Herbal medicine toxicity has been infrequently reported in Spain. We present two cases of acute hepatitis after ingestion of herbal infusions over a period of several months. Among the plants consumed was Teucrium chamaedrys (germander), which has been associated with several cases of hepatotoxicity, mainly in France. After ruling out other causes of hepatitis, we consider that Teucrium chamaedrys provoked the disease. One patient presented acute, cholestatic hepatitis and another presented mixed (hepatocellular and cholestatic) hepatitis. In both patients, the disease was resolved after discontinuing the intake of the herbal teas. We believe that herbal medicine toxicity may be increasing due to the growing consumption of these products. Finally, we emphasize the need to rule out the intake of herbal remedies in patients whose liver tests present abnormalities. PMID- 11412594 TI - [Alendronate-induced hepatocellular lesion]. AB - Bisphosphonates are widely used in the treatment of metabolic bone disease. Bisphosphonate-induced hepatotoxicity is extremely infrequent. We present the case of a 76-year-old female patient with osteoporosis who was prescribed alendronate. A routine laboratory investigation carried out three months later revealed markedly elevated transaminase concentrations. Six weeks after withdrawal of treatment, laboratory parameters had returned to normal levels. We analyze this adverse reaction to alendronate as well as its possible pathogenic mechanisms. PMID- 11412595 TI - [Epidermoid cyst: a rare cause of cystic liver disease]. AB - We present the case of a 42-year-old male patient with a large hepatic cyst, simulating a hydatid cyst. Subtotal cystectomy was performed. Thirteen months after this procedure the patient relapsed and a second laparotomy and total cystectomy were performed. A histopathological diagnosis of epidermoid cyst of the liver was made. The differential diagnosis of epidermoid cysts of the liver is broad and definitive diagnosis is usually only obtained after pathological examination of the surgical specimen. For this reason, and because of the potential for neoplasia, complete resection of the hepatic lesion should be attempted. PMID- 11412596 TI - [Utility of registries of hepatotoxicity caused by drugs]. PMID- 11412597 TI - [The intestinal barrier: functional disorders in digestive and non-digestive diseases]. PMID- 11412598 TI - [Medical treatment of infected pancreatic necrosis]. PMID- 11412599 TI - [Primary liver lymphoma. Diagnostic and therapeutic implications]. PMID- 11412600 TI - [Recombinant human erythropoietin (rHuEPO) in the treatment of primary hemochromatosis]. PMID- 11412601 TI - [Acute pancreatitis secondary to afferent loop syndrome]. PMID- 11412602 TI - [Ki-1 lymphoma of the skin in a patient with Crohn's disease undergoing treatment with azathioprine]. PMID- 11412604 TI - [Clinical pattern and therapeutic results obtained in Germ-Cell testicular cancer in Spain based on a consecutive series of 1250 patients]. AB - BACKGROUND: Even its low incidence, germ-cell testicular cancer is very relevant due to its presentation at young ages and its potential curability over 90%. Spanish Germ Cell Cancer Group (GG) joins the efforts of 51 different Spanish centres to share their experience on the diagnosis and treatment of these special tumours. PATIENTS AND METHOD: We describe the clinical characteristics and the results of treatment in the first 1,250 patients registered throughout 6 years by the GG. RESULTS: 11% had previous criptorchidism. The most frequent initial local simptomatology was increased testis size (90%). 20% lasted more than six months in receiving the first treatment. Inguinal orquidectomy was done in 95% of patients. 435 cases (35%) were seminoma and 815 (65%)non-seminoma. 19% of seminoma and 78% of non-seminoma produced tumour markers. 75% of seminoma but only 56% of non-seminoma were clinical stage I. Following the IGCCCG prognosis classification,20% of non-seminoma fitted in the poor-prognosis group. Stage I seminoma treatment was surveillance, chemotherapy and complementary radiotherapy in 60, 32 and 6%, respectively. Those features were 65, 35% and none in non seminoma cases. Chemotherapy schedules used in advanced cases were EP for seminoma and BEP or BOMP-EPIin non-seminoma, according to whether the patient was in the good or bad prognosis IGCCCG group. With a median of follow-up in all serie of 30 months, we have obtained a three years overall survival of 98% (CI 95%, 96,4-9,6), whereas non-seminoma patients had a three years overall survival of 94% (CI 95%, 92-96). CONCLUSION: The Spanish germ cell testicular cancer clinical pattern is similar to that registered in other occidental countries. Co operative structures like GG,are able to gather an extensive experience in a short period of time that results in achieving a very high number of cured patients. PMID- 11412605 TI - [Prognostic factors of thyroid function following surgical therapy in Graves Basedow's disease]. AB - BACKGROUND: The goals of this work are to describe late thyroid function and to determine predictive factors of permanent hypothyroidism following surgery in Graves-Basedow's disease. PATIENTS AND METHOD: From 1979 to 1999, 107 patients with hyperthyroidism due to Graves-Basedow's disease underwent subtotal thyroidectomy. We performed life-table analysis and calculated the cumulative incidence of hypothyroidism by means of the Kaplan-Meier's method. Survival (euthyroidism)within patients groups was compared using the Mantel-Cox method. Variables influencing long-term thyroid function were determined by estimating the Odds ratio with a logistic regression model. RESULTS: The probability of euthyroidism among all 107 patients at 240 months was 51.4%.Age, gender, duration of both hyperthyroidism and antithyroid therapy and weight of resected thyroid tissue did not influence the eventual outcome. The weight of thyroid remnant was 5.4 (1.5)g and the conditional logistic regression analysis showed that weight of thyroid remnant was the only variable influencing long-term thyroid function. Hyperthyroidism relapsed in 5 patients. CONCLUSIONS: In our experience,surgery represents a definitive alternative treatment with a risk of hypothyroidism within the first 2 years of 43.9%. The weight of thyroid remnant is the only variable influencing long-term thyroid function. PMID- 11412606 TI - [Insulin secretion and insulin sensitivity in relation to glucose tolerance in a group of subjects at a high risk for type 2 diabetes mellitus]. AB - BACKGROUND: Oral glucose tolerance test was evaluated, as well as, insulin sensitivity/secretion in a group of subjects at risk for type 2 diabetes mellitus. METHOD: Metabolic profile of subjects included in the new category, impaired fasting glucose (IFG), was also evaluated. RESULTS: IFG and impaired glucose tolerance identify different alterations in glucose homeostasis. CONCLUSIONS: Those subjects with IFG are heterogeneous in their oral glucose tolerance. They display defects not only in insulin sensitivity but also in their capacity to compensate for it. PMID- 11412607 TI - [Emotional disorders in the working population: an emerging problem in occupational health?]. AB - BACKGROUND: To know the prevalence of emotional disorders in a working population. SUBJECTS AND METHOD: Workers in four companies: construction (A), automobile components (B), electrical and electronic systems (C) and management services (D). The design of the study was cross-sectional with the GHQ-28 questionnaire. RESULTS: The prevalence fluctuated between 8,4% (A) and 24,0% (D). Sex, age, educational level and occupation showed significance differences in some companies. CONCLUSIONS: The results suggest an important prevalence of emotional disorders that likely increase as there will be more firms of services. PMID- 11412608 TI - [Certifying health related web sites: an impossible necessity?]. PMID- 11412609 TI - [Differences between the Spanish and English versions of the last update of the Helsinki Declaration]. PMID- 11412610 TI - [The detection of mental disorders by physicians who are not psychiatrists: usefulness of the PRIME-MD questionnaire]. PMID- 11412611 TI - [Progressive paraparesia of 2 months of evolution in a 24 years-old woman with a renal transplantation]. PMID- 11412612 TI - [Secondary polycythaemia due to chronic carbon monoxide intoxication in non smokers]. PMID- 11412613 TI - [Antibody determination for the human immunodeficiency virus in pregnant women in the public health care area of Gijon, Spain]. PMID- 11412615 TI - [Isolated multiple intrahepatic arteriovenous fistulas as cause of high output cardiac failure]. PMID- 11412617 TI - [Mortality and quality of life of patients beyond 65 years one year after ICU discharge]. AB - BACKGROUND: Intensive care in elderly patients is a subject of controversy, because they generally present a high rate of mortality and short expectation of life. Due to the increasing life expectancy, more elderly patients will be treated in Intensive Care Unit (ICU) with an increasing consume of resources. The present study considers the mortality and quality of life (QOL) of patients beyond 65 years after ICU, and theirs predictors. PATIENTS AND METHOD: Retrospective study of patients >= 65 years admitted in multidisciplinaire ICU. Mortality and QOL (with modified EuroQOL Instrument) one year after discharge were studied. To determine mortality and QOL one year independent predicting factors, multiple logistic regression models were used. RESULTS: Of 313 patients studied, 95 (30%) died in ICU, 32 (10%) in hospital and 34 (11%) died after discharge. The independent predicting factors of mortality one year after ICU discharge were: organ failure (p < 0.000; odds ratio [OR], 2.9), cardiac surgery (p < 0.0000; OR, 0.15) and respiratory disease (p < 0.01; OR, 2.8). Of the 152 surviving patients, 21% got worse their previous QOL and only 17% were severely discapitated. The independent predicting factors of QOL one year after ICU discharge were: prior QOL (p < 0.0002; OR, 10.2) and age (p < 0.002; OR, 0.09). CONCLUSION: Despite the high one year after ICU discharge mortality rate (51%), 83% of the survivors were able to live independently. Due to dependence between mortality and multiorganic failure during ICU stay and not age, this latter cannot be the determining factor of the care level. PMID- 11412618 TI - [Naltrexone in the treatment of alcoholism. Clinical evolution, safety and efficacy in a sample of 198 patients]. AB - BACKGROUND: Our aim was to verify the safety of naltrexone administered during 6 months to alcohol-dependent patients submitted to detoxification. In addition, we aimed at contributing new data on the course of craving and alcohol consumption in naltrexone treated patients and to study prognostic factors predictive of response to treatment. PATIENTS AND METHOD: Multicenter, prospective, pharmacovigilance, follow-up study of a cohort of 198 patients older than 18 years of age meeting DSM-IV criteria for alcohol dependence, who were administered naltrexone over a 6 months period. RESULTS: 77 patients (39.1%) had some adverse event (AA) during the study, including nausea/vomiting (13.7%), sleepiness/sedation (12.8%) and asthenia-laxity-fatigability (10.3%). Only 3.6% of AA were considered severe (6 cases). As to laboratory values, no relevant hematological abnormalities were observed, yet significant reductions from baseline were recorded for cholesterol, total and conjugated bilirubin, transaminases (AST/ALT), and *-GT. Overall, patients had a notable improvement, as reflected by a decrease in craving (from 22.5 to 5.3 on the OCDS scale; p < 0.01), as well as an improved in both global activity (from 68.7 to 85.4 on the EEAG; p < 0.01) and depressive symptoms (from 12.0 to 4.1 on the MDRS scale; p < 0.01). Time passed until first relapse was 33 days, and the abstinence cumulative duration was 126 days. No predictive model of therapeutic success with sufficient statistical power was obtained to guarantee its use in clinical practice. CONCLUSIONS: Naltrexona has a good safety profile to treat alcoholic patients. No predictive factors of response to treatment was found. Thus, the question about which alcoholic patients could mostly benefit from naltrexone treatment remains unanswered. PMID- 11412619 TI - [Effect of vertebral fracture on health related quality of life in a Spanish population older than 54 years]. AB - BACKGROUND: Effect of vertebral fracture on the perceived health using the SF-36 Health Questionnaire in a representative population older than 54 years. SUBJECTS AND METHOD: Randomly cohort from the register of the city Hall of Oviedo. All the 299 subjects (147 men and 152 women) completed the traduced and validated Spanish SF-36 questionnaire four years after radiologic studies were performed to evaluate prevalent vertebral fractures. RESULTS: Vertebral fracture decreased the health related quality of life, particularly in physical function dimension in males and in mental health dimension in women. This effect was increased when osteopenia was present. CONCLUSIONS: This first study performed in both sexes shows worse perceived health in people with fractures. PMID- 11412620 TI - [Medical decisions in patients with chronic obstructive pulmonary disease]. AB - BACKGROUND: A right information about illness is essential if a patient has to take ethically autonomous medical decisions. We assessed whether a group o patients with chronic obstructive pulmonary disease (COPD) had enough information about their illness to take medical decisions. PATIENTS AND METHOD: Structured interview about disease knowledge, wills to receive medical information and to take part in medical lesions as well as opinion on advance directives. RESULTS: Seventy-eight patients with COPD were interviewed. Ninety-two per cent of them were aware that they had a chronic illness and 63% were aware that it was an evolving disease. Fifty per cent believed they could require ICU admission, yet only 33% had knowledge of what mechanical ventilation was. A previous ICU admission and receiving home oxygen therapy were the only factors related to a better knowledge of disease. Forty-six per cent of patients interviewed felt not to be rightly informed, yet 64% of them did not wish to receive more information. Most patients would wish to have some form of future planning of their medical care in case of incompetence. CONCLUSIONS: Patients with COPD do not have enough information to take autonomous decisions. Knowledge is mainly provided by an earlier experience with illness and its complications. PMID- 11412621 TI - [Hypovitamonosis D in the elderly]. PMID- 11412622 TI - [Current in vitro sensitivity of the most frequent bacterial respiratory pathogens: clinical implications]. PMID- 11412623 TI - [Migraine: diagnosis and treatment]. PMID- 11412624 TI - [Functional decline after hospitalization for cardiac insufficiency and chronic obstructive pulmonary disease in the elderly]. PMID- 11412625 TI - [The immigration in the Canary Islands]. PMID- 11412626 TI - [Nonagenarian patients in the emergency department]. PMID- 11412627 TI - [Acute presentation of Sheehan's syndrome. Selective involvement of pituitary stalk]. PMID- 11412628 TI - [Balloon counterpulsation in cocaine induced cardiomyopathy]. PMID- 11412630 TI - [Renal Doppler ultrasonography and its relationship with the renal function in patients with liver cirrhosis]. AB - BACKGROUND: To relate the renal hemodynamic changes, as assessed by Doppler ultrasonography,with the development of ascites, renal function, and endogenous vasoactive systems in patients with liver cirrhosis. PATIENTS AND METHODS: 60 cirrhotic patients were studied prospectively, 31 of these compensated and 29 with ascites. The renal resistive index, renal function and plasmatic levels of renin, aldosterone, noradrenaline and ADH activity were determined. RESULTS: The renal resistive index was significantly higher in the cirrhotic patients with ascites (0.68) than in the compensated cirrhotics (0.63) and was significantly correlated with the serum levels of creatinine,urinary excretion of sodium, plasmatic renin activity and plasmatic concentration of aldosterone. CONCLUSIONS: The renal resistive index, study by means of Doppler ultrasonography, shows progressively increased levels with the evolution of the disease, with the deterioration of the renal function and with the activation of the endogenous vasoactive systems. PMID- 11412631 TI - [Validity of the Spanish version of the Pediatric Quality of Life Questionnaire for evaluating quality of life in asthmatic children]. AB - BACKGROUND: To analyze the clinical utility of the quality of life questionnaire for asthmatic children (PAQLQ) and to validate it for use in clinical practice. PATIENTS AND METHOD: 1,012 children between 6 and 14 years of age, with a diagnosis of mild to moderate asthma attending 48 Spanish hospitals were included in the study. The patients' socio-demographic and clinical characteristics were recorded and all patients were administered the PAQLQ and EQ-5D questionnaires on two occasions: at the baseline visit and at 2-3 months from baseline in patients with uncontrolled asthma(group A) and at baseline and 15 days from baseline in patients with controlled asthma (group B). The feasibility, validity, reliability and sensitivity to change of the PAQLQ were assessed. RESULTS: The PAQLQ proved to be feasible for use in children over 7 years of age with mild or moderate asthma. The PAQLQ did not show a statistically significant relationship with socio-demographic variables, nor with the majority of clinical variables, with the exception of asthma severity, number of exacerbations, symptoms and use ofon- demand short-term beta2 agonist drugs. Correlations between PAQLQ and EQ-5D dimensions were strongest between dimensions measuring similar attributes. Cronbach's *coefficients for the PAQLQ ranged from 0.88 for the limitations in activities dimension to 0.96 for the overall score. The intraclass correlation coefficient for PAQLQscores in group B ranged from 0.71 (limitation of activities)to 0.83 (overall score). The effect size between both visits ranged from 0.49 to 0.69. CONCLUSIONS: The Spanish version of the PAQLQ proved to be valid for use in children with mild to moderate asthma. PMID- 11412632 TI - [Prevalence of microalbuminuria and association with cardiovascular risk factors in a Spanish Mediterranean area]. AB - BACKGROUND: The primary objective of this study was to know the prevalence of microalbuminuria (mAlb)in a general population from the Spanish Mediterranean area along with its association with other cardiovascular risk factors (CRF). PATIENTS AND METHOD: We studied 99 consecutive patients who were classified in mAlb+ or mAlb-,depending on whether their mAlb was higher or lower than 30 mg/day,respectively. RESULTS: Thirty-seven patients(9.3% of the general population) were mAlb positive. Twenty-one percent of them were males with hypertension. The presence of mAlb was positively correlated with age, systolic blood pressure,body mass index, waist-to-hip ratio, pulse pressure, HDL cholesterol,triglycerides, glycemia and serum creatinine. The level of mAlb was 12.9 (11.2) (mg/day) in non-hypertensive non diabetic individuals,while it was 14.1 (16.5) in diabetic non-hypertensive subjects, 22.5 (53,5) in hypertensive non-diabetic subjects and 27.7 (49.1)in diabetic individuals who were also hypertensive. Hypertension correlationed with the presence of mAlb with an odds ratio of 2.4 (CI 95%: 1.1-5.2)and the addition of a male gender increased the odds ratio to 3.8 (CI 95%: 1.8-8.1). CONCLUSIONS: The prevalence of mAlb in the Spanish Mediterranean area appears to be low (9.3%),although it is especially high in hypertensive males with or without diabetes. PMID- 11412633 TI - [Pediatric quality-of-life]. PMID- 11412634 TI - [Allogeneic stem cell transplantation with reduced-intensity conditioning regimens. Another step towards curing by immunologic methods]. PMID- 11412635 TI - [Hematopoietic allogeneic transplantation with attenuated conditioning regimen on an outpatient basis. First experience in Spain]. AB - BACKGROUND: Allogeneic hematopoietic transplantation using attenuated conditioning regimens seems promising. This procedure associates to relatively low morbidity and mortality. In consequence, an outpatient management of this transplantation modality may be considered, even in elderly patients. CLINICAL REPORT AND RESULTS: This approach was considered in a 62 years-old female suffering from chronic myeloid leukemia in chronic phase. The conditioning regimen included fludarabine and 200 cGy of total body irradiation. Cyclosporine A and mycophenolate mofetil were used as immunosuppression. Conditioning, peripheral-blood stem-cell infusion, and postransplant follow-up was managed in the outpatient setting. Two short admissions were required. Eight months after transplant, the patient remains in sustained haematological remission with complete donor chimerism,has a 100% Karnofsky score and continues being managed on an outpatient basis. CONCLUSIONS: Allogeneic stem-cell transplantation can be performed safely on an outpatient basis,even in elderly patients. PMID- 11412636 TI - [In utero hematopoietic stem cell transplantation: current status and future potential]. PMID- 11412637 TI - [Measurement of body fat mass in vivo: from two-compartment techniques to neutron activation analysis and DXA]. PMID- 11412638 TI - [Cholesterol levels in children: a matter of confusion]. PMID- 11412640 TI - [Interpretation of the Paul-Bunnell test]. PMID- 11412641 TI - [Compassionate use protocols and committees on ethics]. PMID- 11412643 TI - [Sporadic and familial Parkinson's disease: comparative study]. AB - BACKGROUND: Several studies have shown that 13 to 33% of patients with Parkinson's disease (PD) exhibit a positive familial history. The goals of this work were to identify patients with familial PD and to analyse whether there existed distinctive features between familial and sporadic cases. PATIENTS AND METHOD: 402 patients with PD from the Hospital Clinic i Universitari of Barcelona were evaluated prospectively. Clinical assessment was done using different scales in 169 patients. The disease was classified as tremorigenic, rigid or mixed according to the predominant symptoms. RESULTS: The frequency of familial PD was 13%. The age at onset was not different between familial and sporadic cases but it was significantly higher in females (57.4 [13] years) than in males (54.8 [11.4] years) (p < 0.05). The tremorigenic type of PD was more common in familial cases (35.5%) (p < 0.05). In familial PD cases, the age at onset was lower in descendents (53 [13] years) than in parents (68 [7.8] years) (p = 0.001). CONCLUSIONS: Genetic factors may play an important role in the development of PD and gender-associated factors may modulate the age at onset. Familial PD cases differ from sporadic cases in the higher frequency of predominantly tremorigenic forms. The lower age at onset in descendents than in parents suggests the existence of a genetic anticipation phenomenon in familial PD. PMID- 11412644 TI - [Trends in cancer mortality in Catalonia (1975-1998)]. AB - BACKGROUND: Aimed at assessing recent trends of cancer mortality in Catalonia (Spain), we analysed mortality rates for main tumour locations in men and women between 1975 and 1998. MATERIAL AND METHOD: Mortality data were obtained from the Mortality Registry of Catalonia. For each sex, age-standardized (standard world population) mortality rates were determined by means of the direct method, and generalized lineal models with Poisson distribution were adjusted to obtain annual change percentages. RESULTS: In men, the overall cancer mortality (except skin non-melanoma) was highest within 1991-1992 (age-standardized rate: 181.39/100,000) and decreased thereafter until 169.47/100,000 in 1995-1996. Lung cancer was the main cause of cancer mortality, displaying a 4% annual increase until 1989-1990, and a subsequent stabilization (-0.5% annual between 1989 -1990 and 1997 - 1998); Colorectal cancer was the second cause of cancer death, both in men and women, with a mortality rate stabilization since the 1990s. Prostate cancer became the third cause of cancer death in 1997-1998. In women, overall cancer (except skin non-melanoma) mortality rate was of 77.98/100,000 in 1997-98, showing a 0.9% annual decrease since 1989-1990. Breast cancer was the chief cause of cancer death within 1997-1998: after a sustained increase since 1975-1976, mortality started to decline (-2.0% annual) since 1989-1990. Lung cancer became the third cause of cancer death among women within 1997-1998, just after breast cancer and colorectal cancer, while gastric and uterine cancer became the fourth and fifth cancer death causes during this period, respectively. CONCLUSION: After years of a steady increase, mortality rates for main cancer types stabilized since 1989-1990 in Catalonia. PMID- 11412645 TI - [Comparative study of piperacillin/tazobactam versus imipenem/cilastatin in febrile neutropenia (1994-1996)]. AB - BACKGROUND: We aimed at comparing the effectiveness and safety of piperacillin/tazobactam(PIP-TAZ) versus imipenem/cilastin (IMI) administered as empiric monotherapy in patients with febrile neutropenia. PATIENTS AND METHOD: Patients with hematological diseases who were randomly assigned either PIP-TAZor IMI were enrolled in the study. A sequential strategy of antibiotic therapy addition was applied as long as fever persisted or microorganisms were isolated at 72 h. Moreover, if bacteriologically unconfirmed fever persisted after 5-7 days, an antifungal therapy was started. The treatment was considered successful if fever and clinical signs resolved and/or pathogens were cleared without adding further antibiotics at 72 h. Differences between percentages were analyzed using the *2test. RESULTS: 137 patients were evaluated. The successful response rate of PIP-TAZ after 72 h was similar to IMI (32.2 and 35.2%). The defervescence time was shorter (3.6 and 4.2 days) and the bacterial response more favourable with PIP-TAZ than with IMI, but statistically significant differences were not reached. The overall response in both groups was 91%.18.2% of episodes were bacteriologically confirmed. The most frequent isolated microorganism was Staphylococcus coagulase-negative(48.8%). There was one only case of septic shock, within the IMI group, and the overall mortality of the group was 8.7%. The occurrence of vomiting in the IMI group was significantly higher than in the PIP TAZ group (39.9 and 5.6%; p < 0.0001). CONCLUSIONS: PIP-TAZ is as effective as IMI and it constitutes a good choice as an initial empiric monotherapy of febrile neutropenia. PMID- 11412646 TI - [Sexual behavior risk in Spanish international travelers]. AB - BACKGROUND: To know the sexual behavior of Spanish international travelers and its association with geographical destinations and sexually transmitted disease acquisition. PATIENTS AND METHOD: 1,008 consecutive patients who attended a tropical out-patient clinic during 26 months were surveyed by means of a previously designed clinical questionnaire that included specific questions regarding sexual practices during the trip. RESULTS: 19% of travelers had sexual intercourse; 53.6% of them having employed a condom. There were no differences regarding gender or destination. 3.4% of travelers who had unprotected sexual intercourse acquired HIV. CONCLUSIONS: A high proportion of travelers have unprotected, risk sexual contacts. A low but alarm PMID- 11412647 TI - [Genetics in Parkinson's disease: toward a new nosological era]. PMID- 11412648 TI - [Introduction to functional food]. PMID- 11412650 TI - [Systematic review of clinical trials on the treatment of rheumatoid arthritis with tumour necrosis factor alpha inhibitors]. PMID- 11412651 TI - [Role of antioxidants in the prevention of cardiovascular disease]. PMID- 11412652 TI - [Hodgkin lymphoma associated with primary Sjogren's syndrome]. PMID- 11412653 TI - [Edimburgh Declaration: something more than an amendment to the Helsinki Declaration]. PMID- 11412654 TI - [Is it necessary to add a fourth drug to the initial treatment of tuberculosis?]. PMID- 11412655 TI - [Is it necessary to add a fourth drug to the initial treatment of tuberculosis?]. PMID- 11412656 TI - [Is it necessary to add a fourth drug to the initial treatment of tuberculosis?]. PMID- 11412657 TI - [Low-molecular-weight heparins in unstable angina and non-Q-wave myocardial infarction]. PMID- 11412660 TI - [CagA and VacA cytotoxin antibodies and risk for peptic ulcer disease in patients with Helicobacter pylori infection]. AB - BACKGROUND: The presence of cagA antibodies constitutes a serum marker of infection caused by virulent strains of Helicobacter pylori. The objective of this study was to determine the risk of peptic ulcer in patients with H. pylori infection, in relation to the detection of CagA and VacA antibodies. PATIENTS AND METHOD: Prospective case-control study including 104 peptic ulcer patients with active H. pylori infection (positive urease test and/or histology, or positive urea breath test) and 104 age- and sex-matched controls, without peptic ulcer history, with active H. pylori infection (positive urea breath test). Serum CagA and VacA antibodies were determined by Western blot. Non-steroidal antiinflamatory drugs (NSAID) use was determined by structured data collection. A multivariate analysis (logistic regression) was carried out to determine the odds ratio (OR). RESULTS: Presence of serum antibodies against CagA was higher in peptic ulcer patients (74%) than in controls (46.2%) (OR = 5.7; 95% CI = 2.1 15.6). However, presence of serum antibodies against VacA in patients (46.2%) was similar to that in controls (36.5%). NSAID use was also more frequent in patients (51.9%) than in controls (21,2%) (OR = 6.5; 95% CI = 2.2-19,5). CONCLUSIONS: Serum antibodies against CagA and use of NSAID are the most important risk factors for peptic ulcer disease. PMID- 11412661 TI - [Diagnostic criteria of the sustained weight test for the detection of arterial hypertensive patients]. AB - BACKGROUND: Like other exercise tests, the sustained weight test (SWT) is based on the cardiovascular reactivity to physical exercise. However, the validity, feasibility and reproducibility criteria inherent to this test are not well defined. Our goals were to describe the systolic arterial pressure (SAP) and diastolic arterial pressure (DAP) values which allow a diagnosis of hypertension with this test and to validate the criteria for diagnosis of hypertension. SUBJECTS AND METHOD: We designed a transversal study with a sample of 547 people. Hypertensive and normotensive individuals were selected considering 140 mmHg for SAP and/or 90 mmHg for DAP as reference values. Subsequently, the SWT test was applied and values of 140/95 mmHg and 150/100 mmHg were identified to define two normotensive and two hypertensive groups, respectively. Finally, we determined the validity, feasibility and reproducibility of the method. RESULTS: The correlation of DAP and SAP before and after the SWT test was fairly high. Values of sensitivity, specificity, predictive positive value and predictive negative value with the 140/95 mmHg approach were 93.3, 87.9, 71.4 and 97.6%, respectively, while these were 89.6, 95.6, 87 and 96,6% when using the 150/100 mmHg approach, respectively. CONCLUSIONS: The results of this study demonstrate a high correlation between the arterial pressure measured by the SWT method and that measured by the international method. Likewise, we have shown an appropriate validity, feasibility and reproducibility of the SWT test for diagnosis of arterial hypertension in the community. PMID- 11412662 TI - [Hepatitis C virus genotypes. Study of 302 patients coinfected by the human immunodeficiency virus]. AB - BACKGROUND: To study the distribution of different HCV genotypes in HIV-infected patients. PATIENTS AND METHOD: This study was carried out in 302 HIV/HCV coinfected patients who were followed-up at the HIV/AIDS Unit of the Arnau de Vilanova University Hospital of Lleida (Spain). HCV genotypes were determined by Inno-Lipa HCV II technique (Innogenetics, Belgium). RESULTS: 143 patients (51.43%) had a genotype-1, followed by genotype-3 (81 patients; 29.13%), genotype 4 (53 patients; 19.06%), and genotype-2 (one patient; 0.35%). It was not possible to know the genotype in 24 patients (NT). CONCLUSIONS: In our health care area, HCV genotype-1 was the commonest among HIV/HCV-coinfected patients. However, a given HIV-infected patient with HCV antibodies has practically the same probability of having a genotype-1 as a genotype non-1. PMID- 11412663 TI - [Factors associated with inappropriate hospitalization at an internal medicine department]. AB - BACKGROUND: To determine the factors associated with inappropriate admissions in an Internal medicine department. SUBJECTS AND METHOD: We included 1,993 admissions. The justification for the admissions was evaluated using the Appropriateness Evalutation Protocol. RESULTS: 187 (9.4%) admissions were considered inappropriate. The likelihood of an inappropriate admission was higher as long as the patient was younger than 65 years of age (OR, 1.94; (95%) IC, 1.32 2.85) and the admission had been programmed from the outpatients clinic (OR, 10.58; (95%) IC, 2.79-40.1). Clinical diagnosis also influenced the inappropriateness of admissions. CONCLUSION: Age, diagnosis and source of patients determine inappropriate admissions in an internal medicine department. PMID- 11412664 TI - [Suitability and usefulness of hospital admissions]. PMID- 11412665 TI - [Medical training and social needs]. PMID- 11412666 TI - [Pharmacological treatment of fistulae in Crohn's disease]. PMID- 11412667 TI - [Human genome: genomics, genetics and medical applications]. PMID- 11412668 TI - [Influence of triglycerides on the lipid profile of cardiovascular risk]. PMID- 11412669 TI - [Pregnancies in human immunodeficiency virus infection discordant couples]. PMID- 11412670 TI - [Proximal femoral fracture in the elderly: usefullnes of the Tinetti index as indicator of physical rehabilitation]. PMID- 11412671 TI - [Geriatric management of the frail elderly with hip fracture may improve their clinical outcome]. PMID- 11412673 TI - [Passive euthanasia]. PMID- 11412675 TI - [Acute pulmonary histoplasmosis after a journey to Peru]. PMID- 11412677 TI - [Apolipoprotein E polymorphism and coronary disease]. AB - BACKGROUND: Apolipoprotein E (Apo E) plays an important role in atherogenesis. Apo E is polymorphic with three codominant alleles: *2, *3 and *4. Inthis study we evaluated the association between *4 allele and coronary heart disease in hypercholesterolemic subjects from Spain. SUBJECTS AND METHOD: We selected 389 subjects (56% women, mean age 57 years) with hypercholesterolemia who had been on a 6-weeks low fat, low cholesterol diet. Measurement of lipid and lipoprotein concentrations and determination of apoE genotype were carried out in a centralized laboratory. RESULTS: ApoE distribution was as follows: *2/*33%, *3/*3 75%, *3/*4 20%, *4/*4 1% and *2/*41%. Subjects were grouped into *4 (n = 83) or no *4 (n = 303) according to the presence or absence of the *4 allele. Three *2/*4 cases were excluded. Prevalence of coronary artery disease was 15.7% in *4 and 6.9% in no *4 (OR 2.49, 95% CI 1.19-5.22). The relationship persisted significant after correction for age, sex, cardiovascular risk factors and serum concentrations of total cholesterol, HDL-cholesterol and triglycerides by multiple logistic regression analysis (OR2.56, 95% CI 1.03-6.39).Conclusion:In Spain, *4 carriers have a higher prevalence of coronary artery disease than no analysis *4 carriers. PMID- 11412679 TI - [The role of diabetes mellitus in cardiac disease in Spain. The CARDIOTENS Study 1999]. AB - BACKGROUND: We analyse the characteristics of the patients with diabetes and cardiac disease included in the CARDIOTENS 1999 study. PATIENTS AND METHOD: 32,051 outpatients who were seen the same day by 1,159 primary healthcare physicians (79%) and cardiologists (21%) were prospectively registered in a database including demographic and clinical data and therapeutic profile. RESULTS: History of cardiac disease was present in 19% (6,194 patients) of the whole population, and 1,275 of them (20.6%) were diabetics. Hypertension was present in 74% of diabetic patients with cardiac disease. Coronary heart disease (angina pectoris or previous myocardial infarction) was present in 45% of diabetic patients with heart failure. Less than 30% of these patients had blood pressure levels under 130/85 mmHg, as recommended by international guidelines. An LDL-cholesterol level lower than 100 mgrs/dl was observed in only 12% of diabetic patients with coronary heart disease; the mean values of total cholesterol and LDL-cholesterol of these patients were significantly (p < 0.01) higher in those seen by primary healthcare physicians. Less than 40% of diabetic patients with cardiac disease were treated with an angiotensin converting enzyme inhibitor, a therapy which was otherwise used in 50% of diabetic patients with heart failure. A beta-blocker therapy was used in 26% of diabetic patients with coronary heart disease and 39% of them were being treated with statins. CONCLUSIONS: More than 20% of patients with cardiac disease in this study were diabetics. Blood pressure and cholesterol levels recommended by current guidelines were attained in a limited proportion of these patients. The use of drugs with demonstrated prognostic benefit in diabetic patients with heart disease is scarce. PMID- 11412680 TI - [Bacteremia in patients discharged from the emergency unit]. AB - BACKGROUND: To know the clinical and epidemiologic characteristics, the evolution and the rate of adequate empiric antibiotic treatment in adult patients with a bacteremia discovered after discharge from an emergency unit. PATIENTS AND METHOD: During 10 years (1989-1998), we registered the clinical and epidemiological data from patients with a bacteremia detected after discharge from emergency. The results were compared with those of patients admitted to hospital who had a community-acquired bacteremia. RESULTS: 85 patients were analysed (52 males), 45% aged over 70 years. Overall, they represented 13.9% of community-acquired bacteremias detected during the same period. Most frequent source was the urinary tract (69%) followed by an unknown source (12%). E. coli was isolated in 62% of cases. In 16 cases, treatment was considered incorrect. Three patients died. CONCLUSIONS: A non-worthless number of patients with bacteremia are discharged from the emergency unit, and 21% of them receive inadequate treatment. Fatal outcome is rare but avoidable in some cases. PMID- 11412681 TI - [Impact of hospital admissions for pneumonia in a tertiary Spanish hospital]. AB - BACKGROUND: Impact of hospital admissions for pneumonia during 1998 in the Hospital Clinic, Barcelona. PATIENTS AND METHOD: Retrospective information on the hospitalizations for pneumonia was collected. RESULTS: During 1998 a total of 626 adult were admitted with a hospital diagnosis of pneumonia (72% were >/= 65 years). The average length of stay was 10 days. Half of these patients were previously hospitalized during the preceding 5 years. CONCLUSIONS: Hospital admissions due to pneumonia (17 cases per 1.000 hospitalizations) accounted for an estimated cost of 2.3% of the cost of all the hospitalized patients. PMID- 11412682 TI - [Statins: an intriguing nexus between osteoporosis and atherothrombosis]. PMID- 11412683 TI - [Continuing medical education, individual or institutional?]. PMID- 11412684 TI - [Prevalence and clinical significance of hepatitis C virus infection in systemic autoimmune diseases]. PMID- 11412685 TI - [Convulsions and left frontotemporal lesion in a 20-year-old woman]. PMID- 11412686 TI - [Language committees development in medical publications: a proposal]. PMID- 11412687 TI - [Lack of agreement between home self-determination and ambulatory monitoring of blood pressure]. PMID- 11412688 TI - [Fierce diagnosis and treatment]. PMID- 11412689 TI - [Acetylsalicylic acid and acute coronary syndrome]. PMID- 11412691 TI - [Primary Sjogren's syndrome: clinical and immunological characteristics of 114 patients]. AB - BACKGROUND: To analyze the clinical and immunological characteristics of a series of 114 patients with primary Sjogren's syndrome (PSS), and to evaluate the different diagnostic criteria and the association to lymphoproliferative disorders. PATIENTS AND METHOD: We included 114 patients (108 female and 6 male) with a diagnosis of PSS. All patients fulfilled the 1993 European Community criteria for the diagnosis of PSS and 76 patients fulfilled the San Diego Criteria. RESULTS: Mean age was 51 years with a mean follow-up of 7.3 years. The commonest clinical manifestation at onset (70%) was xerostomia/xerophtalmia (sicca syndrome). Extra glandular involvement was articular in 42% of cases, neurologic (35%), respiratory (21%) and hepatic (13%). Eleven patients (9%) developed vasculitis, and three (2%) developed a lympho-proliferative disorder. No statistically significant differences regarding symptoms at onset, frequency of glandular or extra glandular manifestations and severity of disease were observed between the two diagnostic criteria groups. HCV infection was associated with vasculitis (p < 0.001; OR: 20.6; CI 95%, 3.2-129) and lymphoproliferative disorders (p < 0.001). CONCLUSIONS: The clinical evolution of PSS does not vary when using different diagnostic criteria (San Diego and European Community criteria). A subset of patients with vasculitis and lymphoproliferative diseases is found to have an associated HCV infection. PMID- 11412692 TI - [Mortality by social and economic characteristics: The Mortality Study of the Autonomous Community of Madrid]. AB - BACKGROUND: To estimate the effect of marital status, household size, employment status, educational level and occupation on mortality. SUBJECTS AND METHOD: About 3,100,000 persons 24 years and older resident on first of May, 1996 in Autonomous Community of Madrid. For the next 19 months information on individuals who died were obtained by linking Mortality Register and 1996 Population Census. RESULTS: Except for household size, the effect on mortality of characteristics analysed was higher in individuals aged 25 to 44 years than in other group of age. In general, married persons had the lowest mortality, except for men older than 64 years where the lowest mortality was found in the never married category. Men aged 45 to 64 years who lived alone had higher mortality than those who lived with other people, while mortality in people aged older than 64 years increased with household size. Inactive persons had higher mortality than active people. Higher education level and higher skilled occupation were associated with lower mortality, except for women aged 45 to 64 years. CONCLUSIONS: This study has identified population groups associated with increased risk of mortality. Monitoring the trend of mortality in those groups will make possible to found the rational basis to implement social and health programmes. PMID- 11412693 TI - [Albumin-to-creatinine ratio as a diagnostic tool for type 2 diabetic nephropathy]. AB - BACKGROUND: Given the low effectiveness of 24 hours determination of urinary albumin excretion (UAE) for the diagnosis of diabetic nephropathy in primary care, we aimed at evaluating the albumin: creatinine ratio (ACR) in the first voided morning urine sample as a diagnostic tool in this setting. PATIENTS AND METHOD: 214 patients with type 2 diabetes were studied. ACR and UAE were compared with correlation and lineal regression analyses. A receiver operating characteristic (ROC) curve analysis was performed to determine the discriminatory value for diagnosis of diabetic nephropathy. RESULTS: The correlation coefficient between ACR and UAE was 0.93 (p<0.01) and the ROC curve analysis showed a value of 2.4 g/mol for women and 1.4 g/mol for men. CONCLUSIONS: ROC curve analysis reveals that ACR is a useful method for diagnosing diabetic nephropathy, although the discriminatory value depends on sex. PMID- 11412695 TI - [Genetics and disease]. PMID- 11412694 TI - [Simple things can also be important]. PMID- 11412696 TI - [Pneumonia in patients with chronic lymphocytic leukemia. Study of 30 episodes]. AB - BACKGROUND: To analyse the etiology, diagnostic methods and response to therapy in 30 episodes of pneumonia diagnosed in 17 patients with chronic lymphocytic leukemia (CLL) between 1995 and 2000. PATIENTS AND METHOD: In each episode of pneumonia the following data were analysed: age, gender, treatment of CLL, antiinfectious prophylaxis, granulocytopenia, CD4/CD8 lymphocytes ratio, hipogammaglobulinemia, origin of pneumonia (nosocomial or community-acquired), localisation, respiratory insufficiency, need for mechanical ventilation, antimicrobial therapy and response. Diagnostic methods included blood and sputum cultures, fiberoptic bronchoscopy and search for antigens in urine (Legionella pneumophila serogroup 1, galactomannan, and Streptococcus pneumoniae). RESULTS: Median age of the series was 60 yr. (range 50-86) and 12 patients were male. Chlorambucil and prednisone were used in 13 cases and fludarabine in 8. Granulocytopenia was present in 14 episodes, hypogammaglobulinemia was seen in 22 and CD4/CD8 ratio was lower than 1 in 8 out of 14 evaluable cases. Etiology of pneumonia was established in 16 episodes (53%). Fiberoptic bronchoscopy was the most useful technique (83% of positive diagnoses) followed by blood cultures (38%). Two patients were diagnosed of aspergillosis at autopsy. Pneumococcus was the most frequent agent (5 cases) followed by Pseudomonas aeruginosa (4), Pneumocystis carinii (2) and Aspergillus fumigatus (2). One out of the two patients with P. carinii pneumonia had received fludarabin and the remaining was treated with prednisone for long time. Ten patients (30%) had died: P. aeruginosa (3 cases), P. carinii (2), A. fumigatus (2), Mycobacterium xenopi (1), and unknown microorganism (2). CONCLUSIONS: In this series of CLL patients the frequency of etiologic diagnosis of pneumonias was good. Pneumococcus was the most frequent microorganism. Pneumonias caused by opportunistic microorganisms were associated to the treatment with fludarabin or prednisone and were associated to a high mortality rate. PMID- 11412697 TI - [Algorithmic models selection in biomedical applications of multiple regression]. PMID- 11412698 TI - [Lipoprotein (a): physiopathology and clinical and therapeutic considerations]. PMID- 11412699 TI - [Etiopathogenesis of Sjogren's syndrome]. PMID- 11412700 TI - [Antibiotics for preterm and premature rupture of amniotic membranes]. PMID- 11412701 TI - [Hepatoblastoma in an adult]. PMID- 11412702 TI - [Nephrotic syndrome and lithium therapy]. PMID- 11412703 TI - [Agreement between home self-determination and ambulatory monitoring of blood pressure]. PMID- 11412704 TI - [Prolactin and polycystic ovary syndrome]. PMID- 11412706 TI - [Out patient neurologic disease in Spain]. PMID- 11412707 TI - [Inverted metamorphopsia: an alteration in the perception of a special situation of objects]. AB - OBJECTIVES: This study describes the characteristics of six patients who consulted because of episodes of perceptive alteration of extrapersonal space consisting in an inversion of the situation of objects although without changes in shape or size (inverted metamorphopsia). PATIENTS: Six patients (4 males and 2 females) who presented episodes of inverted metamorphopsia were studied: 4 referred right-left inversion (axial plane), 1 upwards-downwards (saggital plane) and 1 inclination of environment (clockwise movement). None had any alteration in the perception of their own bodies during the episodes which lasted from 3 minutes to 1 hour, reporting from 1 to 40 episodes. The most notable antecedents were: traumatism and forced cervical posture (2), arterial hypertension (3), migraine (2), hyperlipemia (2) and protuberantial ischemia (1). RESULTS: Intercritical neurologic exploration was normal. MR study showed leukoaraiosis (4), protuberantial paramedian infarctions (1) and was normal in one case of basilar migraine. Study of vestibular function (performed in 4 cases) did not show alterations. Three patients received antiepileptic drugs (1 carbamazepine and 2 gabapentin) with favorable response. CONCLUSIONS: Inverted metamorphopsia is a paroxistic phenomenon which transduces a dysfunction of the posterior parietal cortex or its afferences (propioceptive, vestibular, retinian). The causes may be multiple (traumatism, migraine, ischemia). Antiepileptic drugs may have a therapeutic role in the cases with multiple episodes. PMID- 11412708 TI - [Epidemiology of ambulatory neurological diseases at the Baix Camp]. AB - BACKGROUND: Studies analyzing the incidence of ambulatory neurological diseases are scanty. INTRODUCTION: To describe the first prospective study of incidence of ambulatory neurological diseases at the Baix Camp (Tarragona). PATIENTS AND METHODS: This is a prospective study of a cohort of consecutive, first visited patients at the neurology clinic of the Sant Joan Universitary Hospital of Reus. Demographic and clinical variables were analyzed. RESULTS: There were 1330 first visits, but 52 were excluded as they were diagnosed and treated previously at another medical center. So, the final study group included 1278 patients; 790 (61.8%) were woman and 488 (38.2%) men. Mean age was 48.7 years. The incidence ratio was 10.3 new neurological outpatients by 1000 hab./year. Headache was the most frequent symptom (36.5%), followed by focal symptoms and transitory loss of consciousness. The most frequent diagnoses were, by order of frequency, non tumoral headaches (35.8%), followed by anxiety and depression, neuropathies and epilepsy. Cranial CT and MRI were the complementary exams more solicited. CONCLUSIONS: This epidemiological study corroborates that the most frequent neurological diseases were non-tumoral headaches, neuropathies and epilepsy. The need for specialized assistance could be reduced by better identification of certain diseases from primary medical assistance. This analysis will be actualized every five years. PMID- 11412709 TI - [Alpha-synucleinopathies]. AB - The term alpha-synucleinopathy is used to name a group of disorders having in common the abnormal deposition of alpha-synuclein in the cytoplasm of neurons or glial cells, as well as in extracellular deposits of amyloid. In Parkinson's disease and Lewy body dementia, alpha-synuclein is the main component of Lewy bodies and dystrophic neurites; alpha-synuclein also accumulates in the cytoplasm of glial cells. In multiple system atrophy, alpha-synuclein conforms the cytoplasmic oligodendroglial inclusions and the neuronal inclusions which are the hallmark of this disease. Finally, the amyloidogenic fragment 61-95 amino acids of alpha-synuclein is the non-Abeta component of senile plaque amyloid in Alzheimer disease. Accumulations of alpha-synuclein in all these disorders have in common a fibrilar configuration, but they differ in the binding of alpha synuclein to distinct proteins with the exception of ubiquitin whose binding to alpha-synuclein is common to all alpha-synuclein inclusions. The mechanisms leading to alpha-synuclein fragmentation and aggegation into extracellular amyloid are not known, although alpha-synuclein fragment and betaA4 aggregates are the result of abnormal cleavage of large precursors. On the other hand, several studies have shown that alpha-synuclein may adopt a fibrilar conformation and give rise to insoluble forms and high molecular weight aggregates in vitro. Similar complexes have also been observed in alpha-synucleinopathies. Although studies in vitro and in vivo have shown toxic effects of alpha-synuclein, the consequence of alpha-synuclein deposition on cell survival in alpha synucleinopathies is not known. PMID- 11412710 TI - [Spinal paresia: semiologic notes]. PMID- 11412711 TI - [Spastic paraparesias by HTLV-1: early identification of a new case. Review of the Spanish casuistics]. AB - This is a study of the early identification of a new case of tropical spastic paraparesis/HLTV-1-associated myelopathy in a Spanish patient not previously reported on. The patient is 48 years of age, and he has been living in Sierra Leone for the past seventeen years, where he has worked as a surgeon. On first examination, he had been suffering from symptoms of a spastic paraparesis for the previous five months. This diagnosis was confirmed using serological tests (EIA and Western-blot) on the blood and cerebrospinal fluid (CSF). We have studied the cases outlined in medical literature, paying special attention to the time elapsed between the onset of symptoms and the date on which the diagnosis was made. PMID- 11412712 TI - [Unilateral optic neuropathy associated to a dural arterio-venous fistula]. AB - We report the case of a 43-year-old female patient that had a subacute loss of visual acuity on her left eye, initially lacking any additional symptom or sign of intracranial hypertension. She was diagnosed and studied as an optic neuropathy. The cranial MR was normal and it did not show changes on the signal of the optic nerve. The patient did not improve with steroidal treatment. She was re-admitted three months later due to cephalalgia without any modification of the visual symptomatology. On this occasion a high intracranial pressure (400 mmH20) was recorder on lumbar tap. A thrombosis of the right transverse sinus with an associated complex dural arterio-venous fistula, was visualized at the Angio-MRI and cerebral arteriography. We suggest a relationship between optical neuropathy and dural arterio-venous fistula. We also discuss the attitude with regard to patients suffering from optic neuropathies and endocranial hypertension of uncertain origin. PMID- 11412713 TI - [Intradural lumbar disk hernia]. AB - Intradural disc herniation is a rare complication of degenerative disc disease. A correct diagnosis of this process is frequently difficult. If this entity is not preoperatively diagnosed and is omitted at surgery, severe neurologic sequels may be provoked. We report a case of a pathologically proven intradural disc herniation preoperatively diagnosed by MR imaging. Clinically, it was manifested by sudden onset of right leg ciatalgia and progressive right lower extremity weakness. The patient also referred a one-month history of sexual dysfunction. MR imaging revealed interruption of the low signal of the anulus fibrosus and of the posterior longitudinal ligament at L2-L3 level and a voluminous disc fragment migrated in the dural sac that showed rim enhancement with gadolinium.The clinical, neuroradiological, and surgical management of lumbar intradural disc herniation are reviewed. PMID- 11412714 TI - [Embolization of a giant cavernous aneurysm with panhypopituitarism]. PMID- 11412715 TI - [Neurosarcoidosis and silicosis: an exceptional association. Clinical pathological study of one case]. PMID- 11412716 TI - [Trigeminal neuralgia as the first symptom of Chiari malformation]. PMID- 11412717 TI - [Myofibrillar myopathies: another complex myopathic syndrome]. PMID- 11412718 TI - [Familial myopathy with desmin storage seen as a granulo-filamentar, electron dense material without mutation of the alphabeta-crystallin gene]. AB - BACKGROUND: Desmin-related myopathy is a familial myopathy and cardiomyopathy. Three subgroups have been outlined, an autosomal dominant (AD) granulofilamentous type with cardiomyopathy, an AD cytoplasmic/spheroid inclusion body type and an autosomic recessive Mallory body-like inclusion type. Recently, in one family belonging to the first group it has been identified a mutation within a gene coding for a chaperone protein, alphabeta-crystallin (CRYAB gene). OBJECTIVE: To describe a Spanish family with a desmin-related myopathy, an AD granulofilamentous type with cardiomyopathy, with a molecular study that does not express any point mutation in the CRYAB gene. PATIENTS, METHODS AND RESULTS: This report concerns a family from southern Spain in which 2 sisters (37 and 44 and 36 years old respectively) present an involvement of skeletal (distal more than proximal) and velo-pharyngeal muscles with onset at the third decade of life and with a rather severe progression, leading soon to bilateral foot drops. The mode of transmission is autosomal dominant. Two other members, the mother and one brother _the latter with hypertrophic cardiomyopathy, atrioventricular block and severe heart failure_, have already died. The electrophysiological study carried out in the sisters shows myogenic patterns with abundant spontaneous activity but not genuine myotonic discharges. Muscle biopsy, also purchased in both sisters, shows myopathic changes, a few rimmed vacuoles in one of them, and several "rubbed-out" fibers or fibers "effacees" visible within the intramyofibrilary network on the enzymatic stainings. The ultrastructural findings in both biopsies reveal a similar pattern of structural damage: an intrasarcoplasmic (specially subsarcolemmal) accumulation of an electro-dense filamentous material arising from the Z-bands and a focal disruption of the myofibrils. Immunohistochemically the subsarcolemmal material stained positively for the intermediate filament protein desmin but other proteins as ubiquitin, tropomyosine and actin were found overexpressed. Genetic studies have excluded myotonic dystrophy, any point mutation in the CRYAB gene, and different markers failled to express linkage to any loci on chromosome 12. The possible mutations in desmin gene are to be studied. CONCLUSION: The distinct intrafamilial phenotype observed in this family confirms the multisystemic character of the desmin-related myopathyies. The genetic results also support the genetic heterogeneity of these myopathies and reinforce the concept that mutations in the desmin gene must be investigated as a cause of the disease. PMID- 11412719 TI - [The importance of hydrostatic valves in the treatment of adult chronic hydrocephalus]. AB - BACKGROUND AND OBJECTIVE: Earlier designs of hydrocephalus valves, focusing on a reduction of the problem of overdrainage, were proved to be insufficiently. Since the middle of the 90's hydrostatic valve, constructions are available, which should solve this problem. The objective of the study was to evaluate, whether these gravitational shunts would indeed reduce the percentage of overdrainage in shunted patients. METHODS: In 51 patients clinical evaluation and MRI was performed pre- and in regular intervals postoperatively. 21 received a Dual Switch valve, 14 a programmable Codman Hakim plus Shuntassistent and 6 a combination of a conventional valve with Shuntassistent. RESULTS: Over 90% of the patients had clear and persisting (over 24 months) clinical benefit from shunting. Only one suffered for some days after shunt implantation from headache in the upright position. No other clinical or imaging hints were given on an overdrainage. The ventricular size was unchanged in patients with a normal pressure hydrocephalus after shunting. In patients with occlusive hydrocephalus only a few showed a significant reduction of the ventricular wide. CONCLUSION: Gravitational shunts seem to decrease the overdrainage linked to the treatment of chronic hydrocephalus in shunted patients, therefore its use should be standardised when treatment chronic hydrocephalus. PMID- 11412720 TI - [Nightmares and behavior during REM sleep]. AB - This study reports the current neurobiologic data on REM sleep including psychoanalysis, distinguishing between the "instincts of pleasure" and the "instincts of death" (Beyond the principles of pleasure, S. Freud, 1920). The award and punishment systems are also reported and current data related to nightmares and mainly dream behavior during REM sleep without atonia are presented underlining the frequency of parasomnia in ponto-cerebellous atrophy and Parkinson's disease. The author emphasizes the need for establishing discussions among the different neuroscientific fields related to these questions. PMID- 11412721 TI - [Traumatic anterior optical neuropathy]. PMID- 11412722 TI - [Alien hand syndrome in Creutzfeldt-Jakob disease]. AB - Alien hand syndrome is defined by uncontrolable actions of the arm and hand that seem to have a purpose. It is usually associated with acute focal lesions after a stroke or surgery of the corpus callosum. It has been described in chronic dementiating diseases such as cortico-basal degeneration, Alzheimer's disease, orthochromatic leukodystrophy and Marchiafava-Bignami disease. We now report a patient with Creutzfeldt-Jakob disease and alien hand syndrome, which appeared after the cognitive alterations and had characteristics of the frontal type of alien hand affecting the dominant upper limb. In the three cases described previously in the literature the alien hand affected the non-dominant hand and the abnormal movement appeared before dementia was clinically obvious. The non specific nature and poorly localising sign of alien hand is remarked. Alien hand should be added the list of features of Creutzfeldt-Jakob disease. PMID- 11412723 TI - [Ischemic stroke in a patient with factor XII (Hageman) deficiency]. AB - Unlike most other coagulation factor deficiencies, usually associated with abnormal bleeding, lack of factor XII (Hageman) can result in thromboembolic events as a result of a deficient fibrinolytic system. We report a patient with an ischemic stroke and factor XII deficiency, a rare hereditary disorder. The optimal therapy for these uncommon disorders is not well established, but they probably require long term anticoagulation to prevent subsequent thrombotic events. PMID- 11412724 TI - [Proximal neuropathy of the median nerve]. AB - In a 77 year old man the rare clinical picture of a complete lesion of the median nerve at the upper arm is described. During removal of a "neurinoma" at the upper arm, inadvertedly the general surgeon had also transsected the median nerve. However diagnosis was delayed due to electrodiagnostic tests, which erronously interpreted the volume conduction of the neigbouring nerves as partial function, and later even as improvement of the median nerve function. Clinically he presented with "orators" hand. He was unable to pinch thumb and index finger, Sensory loss was noted at the fingertip 2 and 3, atrophy of the abductor pollicis brevis muscle and trophic skin changes with an ulcer at the tip of the index finger. Nerve revision confirmed the median nerve transsection. Intraoperative nerve stimulation could not elicit distal muscle stimulation. Sural nerve transplant was performed and within one year a positive Tinel sign progressing 20 cm distally to the operation site, without distal motor or sensory changes was observed. PMID- 11412725 TI - [Sjogren's syndrome and multiple sclerosis]. AB - INTRODUCTION: Multiple sclerosis (MS) usually follows a relapsing-remitting course and attacks multiple areas of the central nervous system (CNS). Certain rheumatic diseases, including Sjogren's syndrome (SS), can present with a similar clinical picture. RESULTS: Two patients out 67 of 100 with MS exhibited xerophthalmia and xerostomia and positive Ro antibodies, thus fulfilling 55 diagnostic criteria. Case 1. A 62-year-old woman developed several episodes of numbress and weakness in her left extremity and ataxia. MRI demonstrated a high intensity areas in periventricular white matter. Initial laboratory studies failure to demonstrate antinuclear antibodies. Xerostomia and xerophthalmia were apparent. Follow-up laboratory examinations showed elevated ANA, Anti-SSA and anti-SSB. Case 2. A 58-year-old woman was diagnosed as having MS at the age of 53 years. Several high-signal foci on MRI were demonstrated in the white matter. Initial serum findings including autoantibodies were unremarkable. Sicca syndrome was present. Laboratory investigations included elevated anti-SSA, anti-SSARo52 and ANA while anti-SSB was within normal limits. DISCUSSION: Primary SS is a chronic autoimmune inflammatory disease of unknown etiology. The CNS symptoms are present in 20-25% of the patients with SS. Some patients have a relapsing remitting course mimicking MS. Focal brain lesions in SS can occur in the cerebral white matter. The features of our patients sufficiently mimicked those of MS and this disorder was the diagnosis in each patient at the time of initial evaluation. SS should be considered in th PMID- 11412726 TI - [Stroke and scleroderma. Cerebral Raynaud?]. PMID- 11412728 TI - [Paraneoplastic sensory neuropathy due to pharynx and tonsil squamous cell carcinoma]. PMID- 11412729 TI - [Comparison of ropivacaine and bupivacaine for epidural analgesia during labor]. AB - OBJECTIVES: To compare the analgesic efficacy and level of motor block using two local anesthetics, ropivacaine and bupivacaine, during labor. MATERIAL AND METHODS: Sixty nulliparous women were enrolled during labor after full-term pregnancies. They were randomly assigned to receive epidural analgesia with ropivacaine (group R) or bupivacaine (group B). Group R patients received 10 ml of 0.18% ropivacaine with 5 microgram/ml of fentanyl followed by continuous epidural infusion of 0.1% ropivacaine with 2 microgram/ml of fentanyl at a rate of 10 ml/h. Group B patients received 10 ml of 0.15% bupivacaine with 5 microgram/ml of fentanyl followed by continuous epidural perfusion of 0.0625% bupivacaine with 2 microgram/ml of fentanyl at the same rate. Pain intensity was assessed on a visual analog scale, motor blockade on a Bromage scale, and level of sensory block at different moments. We also recorded total doses of local anesthetic employed during continuous epidural infusion, manner of final delivery, Apgar score, degree of maternal satisfaction and side effects. RESULTS: The demographic and delivery characteristics were similar in both groups. We found no statistically significant differences between the two groups for level of motor blockade, which was nil for 29 patients (96.66%) in group R and 28 patients (93.33%) in group B. No differences in degree of pain or level of sensory block (T8-T10 in both groups) were observed. The total doses of local anesthetic used were similar at 23.7 +/- 11.6 mg in group R and 16.5 +/- 7.3 mg in group B (non-significant difference). Nor did we find differences in manner of delivery, neonatal Apgar scores, degree of maternal satisfaction or side effects. CONCLUSION: Ropivacaine and bupivacaine are equally effective for epidural analgesia during labor at the doses used and they do not cause a relevant level of motor blockade. PMID- 11412730 TI - [Sedation in spinal anesthesia. Comparison of remifentanil and propofol]. AB - OBJECTIVES: To assess the efficacy and safety of remifentanil for analgesia and sedation during subarachnoidea anesthesia, and to compare remifentanil with propofol. METHOD: Ninety ASA I-III patients undergoing orthopedic or traumatologic surgery under subarachnoid anesthesia were enrolled for prospective study and randomly assigned to two treatment groups. The propofol group (n = 45) received a single dose of 0.5 mg/kg followed by infusion at 3 mg/kg/h. The remifentanil group (n = 45) received a single dose of 0.5 microgram/kg followed by infusion at 0.1 microgram/kg/min. We evaluated quality of sedation, pain intensity during nerve blockade, hemodynamic and respiratory parameters and time until recovery. RESULTS: The remifentanil group experienced less moderate-to intense pain (13%) than did the propofol group (63%) (p < 0.01). Sedation was adequate in both groups and was easy to control by adjusting the rate of infusion. Times until recovery of consciousness and respiratory frequency after withdrawal of infusion until recovery of baseline levels were 7.87 +/- 3.54 min and 5.22 +/- 2.49 min, respectively, in the remifentanil group and 8.72 +/- 4.59 min and 5.36 +/- 2.49 min, respectively, in the propofol group, respectively. Patients in the remifentanil group experienced a significantly greater decrease in SpO2 than did those in the propofol group (20% and 4%, respectively; p < 0.05). Mean blood pressure was higher for patients treated with remifentanil. The incidence of vomiting was also higher in the remifentanil group than in the propofol group (9% vs 0%). CONCLUSION: Remifentanil is more effective in reducing pain related to nerve blockade and level of sedation is lower; however remifentanil is associated with a higher incidence of respiratory depression and vomiting. PMID- 11412731 TI - [Use of aprotinin during pediatric heart surgery]. AB - OBJECTIVE: To study the efficacy of high doses of aprotinin during cardiopulmonary bypass (CPBP) in pediatric patients with a tendency to intra- and postoperative bleeding: children undergoing repeated operations and those with cyanotic cardiopathy. PATIENTS AND METHODS: A simple retrospective, random sample of 71 pediatric patients with cyanogenic cardiopathy was studied. Group I (n = 36) received a test dose of 50,000 KIU in 2 min. The initial dose was 1,700,000 KIU (170 ml/m2 of body surface) i.v., at an infusion rate of 150 ml/h. CPBP circuit prime dose was 1,700,000 (170 ml/m2 of body surface), with replacement at a volume equivalent to the circuit prime. The maintenance dose was 400,000 KIU (40 ml/m2/h) i.v. Infusion was suspended 2 hours after a patient's arrival at the intensive care unit (ICU). We recorded red blood cell loss and blood product requirements in the first 24 hours in the ICU, duration of surgery, intra- and postoperative diuresis, and complications. RESULTS: Group I required 13.96 +/- 12.09 ml/kg/h of packed red cells whereas Group II required 22 +/- 16.22 ml/kg/h (p < 0.05). The volume lost was less in Group I than in Group II (0.77 +/- 0.48 ml/kg/h vs. 2.12 +/- 3.9 ml/kg/h; p < 0.05). The means for intra- and postoperative diuresis in the first 24 hours were not significantly different. CONCLUSIONS: Aprotinin significantly reduces red blood cell loss and homologous blood transfusion requirements. No complications attributable to aprotinin were observed. PMID- 11412732 TI - [The efficacy and safety of continuous epidural analgesia versus intradural epidural analgesia during labor]. AB - OBJECTIVE: To determine the efficacy and safety of intradural-epidural analgesia in comparison with continuous epidural analgesia during labor and childbirth. PATIENTS AND METHOD: Forty-two women whose labor began spontaneously were enrolled and distributed randomly in two groups. The intradural-epidural analgesia group (IEA, n = 21) received 25 microgram of intradural fentanyl with 2.5 mg of isobaric bupivacaine with adrenalin, after which analgesia was maintained with epidural administration of one 8 mL bolus of 0.125% bupivacaine, followed by perfusion of a balanced concentration at a rate of 8 ml/h. Patients in the continuous epidural analgesia group (CEA, n = 21) were given 8 ml of 0.25% bupivacaine with adrenalin; the epidural perfusion of 0.125% bupivacaine and 1 microgram/ml of fentanyl was started at the same rate as in the IEA group. We recorded pain as assessed on a visual analog scale, extension of sensory and motor block, maternal hemodynamic constants, number of boluses of bupivacaine used, total doses of bupivacaine and oxytocin, instruments needed for childbirth, and side effects (pruritus, nausea and vomiting). RESULTS: Analgesic efficacy during the first 30 minutes was greater in the IEA group. The total dose of bupivacaine, required top-up boluses, and the extension of sensory block at 30 minutes, one hour and two hours were also significantly less in the IEA group. The incidence of pruritus was higher in the IEA group. No significant differences were observed for other variables. CONCLUSIONS: Intradural-epidural analgesia provides effective analgesia for labor, with rapid onset, reduced extension of sensory block, lower total doses of local anesthetics and few side effects. PMID- 11412733 TI - [Guidelines for basic and advanced cardiopulmonary resuscitation of children. Commission for Cardiopulmonary Resuscitation of the Spanish Society for Anesthesiology and Resuscitation]. PMID- 11412734 TI - [Controversies surrounding pain and inhalation anesthesia in nineteenth century Spain]. AB - The introduction of inhaled anesthetics to Spain in 1847 brought appearances by enthusiastic promotors, prudent and reserved admirers and stubborn adversaries - sparking controversy over the use of the new gases. In some cases debate involved the discussion of various concepts of pain, as is shown by heated exchanges among Eusebio Castelo Serra, Manuel Santos Guerra and Zacarias Benito Gonzalez in the pages of the journal Boletin de Medicina, Cirugia y Farmacia, in three articles appearing between 1850 and 1851 on the concept of pain: Sobre el dolor de las enfermedades y principalmente en las operaciones quirurgicas, Modificacion de dos instrumentos and Estudios sobre el dolor. Investigation into the authors' biographies and an analysis of the content of the articles has permitted us to reconstruct some aspects of the concept of pain in Spain in the middle of the nineteenth century. PMID- 11412735 TI - [General anesthesia with remifentanil in two cases of emergency cesarean section]. AB - Remifentanil is a synthetic opiate with evident advantages for various anesthetic techniques, enhancing quality of anesthesia. Indications are increasingly well defined. Remifentanil may be used in obstetric analgesia-anesthesia thanks to advantages demonstrated in patients with heart disease (cardiac and non-cardiac anesthesia) and in those requiring neuroanesthesia. Remifentanil is known to cross the placenta rapidly and to be rapidly metabolized and redistributed to both mother and fetus. Based on this, and on pharmacokinetic and pharmacodynamic studies in children, we judged remifentanil to be indicated for use in two patients undergoing emergency cesarean section, for whom hemodynamic stability and immediate postoperative assessment were basic requirements. The first case involved a woman 40 weeks pregnant with a history of mitral valve prolapse and an episode of acute pulmonary edema in the 28th week, who presented with ruptured membrane and the fetus in sacroposterior breech presentation without subsequent progression of labor. The second involved a woman 40-weeks pregnant with a diagnosis of Hunt-Hess grade II subarachnoid hemorrhage who had gone into labor. Outcome was satisfactory in both cases, with no complications potentially affecting the status of either mother or child. No infant respiratory insufficiency occurred and Apgar scores were favorable. We consider remifentanil to be safe and effective for general anesthesia for emergency cesarean delivery in patients with cardiac and/or neurological risk factors. PMID- 11412736 TI - [Ventricular fibrillation coinciding with the use of an electric scalpel during laparoscopic surgery]. PMID- 11412737 TI - [Hypernatremia in laparoscopic surgery for a hydatid liver cyst in a pediatric patient]. PMID- 11412738 TI - [On the analysis of the cost-effectiveness ratio for autologous transfusion programs]. PMID- 11412739 TI - [On neurophysiologic monitoring during surgery for scoliosis]. PMID- 11412740 TI - [On amniotic fluid embolism]. PMID- 11412742 TI - [Orotracheal tube replacement during surgery using a femoral vein catheter in a patient with a difficult upper airway]. PMID- 11412743 TI - [Generation of new cardiomyocytes in the adult heart: Prospects of myocardial regeneration as an alternative to cardiac transplantation]. AB - The classic dogma, still prevalent in cardiology, that the adult myocardium is a terminally differentiated tissue unable to produce new cardiomyocytes needs to be revised in light of recent results. In human and experimental animals there is now incontrovertible evidence that new myocytes are continuously generated throughout life in response to physiological and pathological stimuli. Moreover, the elucidation of mechanisms responsible for the hypertrophic response indicate similarity and overlap with the mechanisms involved in cell death by apoptosis as well as cell growth. During cardiac development, from birth to adulthood, there is a balance between the stimuli induce cell growth -by hypertrophy and hyperplasia- on one hand and those that induce programmed cell death on the other. In human and experimental animals it has been well documented that pathological conditions, such as diabetes and hypertension, can increase dramatically the rate of cell death. Moreover, high rates of cell death have been measured in normal adult human hearts and those of mice and rats. No surprisingly, these values increase significantly with age and high in senescence. By themselves, these high rates of normal cell death provide a very compelling argument in favor of cardiomyocyte regeneration. Without cell renewal, these rates of cell death would be incompatible with survival because the heart would disappear before early adulthood. As expected, direct measurement of rates of new cell formation in adult hearts demonstrate high rates of cell renewal that compensate for cell death. Thus, the heart is in continuous cellular turnover with new myocardial cells replacing the older ones. Experiments with fetal mouse cardiocytes shows that the retinoblastoma protein is responsible for the cardiocyte withdrawal from the cell cycle during development. The identification in the adult heart of a subpopulation of quiescent cells that have many of the characteristics of stem cells able to rapidly enter the cell cycle and generate new cardiocytes is yet another evidence that the heart continuously produces new cardiocytes to replace those that disappear either by apoptosis or necrosis.Surprisingly, stem cells other that those from the heart are able to produce new cardiocytes and repopulate the myocardium. We have used bone marrow stem cells injected into the border zone of post-coronary occlusion necrosis. Remarkably, these cells have proven to be very effective in generating new myocardium in the necrotic zone that is integrated to the rest of the muscle and irrigated by new vessels. These results demonstrate that stem cells provide a new avenue for the generation of new contractile tissue. This approach could prove useful in the treatment of chronic cardiac failure and post-ischemic necrosis. PMID- 11412744 TI - [Are there clinical implications in the registries of new devices in coronary interventionism?]. PMID- 11412745 TI - [Myocardial revascularization: clinical outcomes and determinants of quality of life]. PMID- 11412746 TI - [In hospital and mid-term outcome of patients with NIR stent implantation: multicenter ESPORT-NIR registry]. AB - BACKGROUND AND OBJECTIVE: Despite improvements in the results and techniques of catheter-based revascularization, few studies have evaluated the clinical results of the application of new stent designs. We describe the in-hospital and mid-term outcome of patients undergoing a stent NIR implantation. PATIENTS AND METHOD: At least 1 Stent NIR was implanted in 1.004 patients (1.136 lesions) recruited from 50 centers in an international, multicenter, prospective, registry (Spain and Portugal NIR stent registry). Inclusion criteria were objective coronary ischemia related to a severe de novo lesion or first restenosis in native vessels with a reference diameter >= 2.75 mm. The primary end-point was the incidence of major adverse cardiac events within the first 7 months of follow-up. RESULTS: The mean age of the patients was 60 years and 82% were male. Angioplasty was indicated due to unstable angina in 61% of the cases. Stent implantation was successfully achieved in 99.6%. Clinical success (angiographic success without in-hospital major events) was achieved in 98.6% of patients. The rate of angiographic restenosis (> 50% stenosis narrowing) was 16% (CI 95%; 11.7-21.2). The accumulated major cardiac adverse event rate at seven months of follow-up was 8.7%: death (0.9%), acute myocardial infarction (1.2%) and target lesion revascularization (6.6%). CONCLUSION: In the wide setting of the population included in the ESPORT-NIR registry, stent NIR implantation was a highly effective therapy with a good mid-term clinical and angiographic outcome. PMID- 11412747 TI - [Initial results and 6 month clinical follow-up after implantation of a silicon carbide coated coronary stent]. AB - INTRODUCTION AND OBJECTIVES: To present the initial Spanish experience with the Tenax coronary stent, a laser sculpted from high-precision 316L stainless steel coated with hydrogen rich amorphous silicon carbide that reduces thrombogenecity and improves biocompatibility. PATIENTS AND METHODS: From July 1998 to July 1999, 206 patients (62 +/- 5 years) underwent implantation of 231 Tenax stents in 9 centers as the only revascularization procedure. The most frequent clinical indication was unstable angina (66%), and most of the lesions were complex (class B2 and C). The target vessels were the left anterior descending (51%) and right coronary arteries (36%). The ejection fraction was < 0.5 in 19% cases. RESULTS: Revascularization was complete in 70%, elective in 80%, and the implantation was direct in 25% of the cases. The procedure was successful in all the lesions, reducing stenosis from 62 +/- 16 to 16 +/- 10% and increasing the minimal luminal diameter from 0.81 +/- 0.40 to 2.61 +/- 0.59 mm. The TIMI flow was reduced in 30%, but normalized after the stent in all but one case. The incidence of cardiac events was minimal: 1 acute thrombosis (0.5%) resolved by a new angioplasty and 1 non-Q myocardial infarction (0.5%). At the 6-month clinical follow-up 10% of the patients presented complaints of angina greater than class II, and a new angioplasty was carried out in 1.9% of these cases. CONCLUSION: Clinical and angiographic data suggest that the hydrogenated silicon carbide coating of the Tenax coronary stent may indeed play a beneficial role in patient outcome, and should therefore be evaluated by prospective clinical trials. PMID- 11412748 TI - [The role of mitral valve balloon valvuloplasty in the treatment of rheumatic mitral valve stenosis during pregnancy]. AB - OBJECTIVE: To analyze the immediate results and the clinical evolution of a group of fertile age women with rheumatic mitral stenosis, in whom percutaneous balloon mitral valvuloplasty was performed before or during pregnancy. PATIENTS AND METHOD: Eighty-one women with mitral stenosis, submitted to balloon mitral vavuloplasty, were studied. They were divided into three groups, according to their desire of no further pregnancies (group A; n = 19), pregnancy during the follow-up (group B; n = 23) or valvuloplasty was performed during pregnancy (group C; n = 39). Patients from group B and C were controlled during pregnancy, childbirth and puerperium, and the newborns of women in group C were followed from birth to the age of 5 years. RESULTS: Mortality in the three groups was null and the incidence of miscarriage was 2 (8.6%) in group B and 3 (9.1%) in group C. Normal delivery was predominant in group B and delivery by caesarean was predominant in group B. Success was immediate in all the cases. The procedure was repeated in 3 women due to restenosis. The media valvar area rase from 0.93 to 2.05 cm2 in group A, from 1.28 to 2.04 cm2 in group B and from 0.84 to 2.14 cm2 in group C (intergroup p = NS). The functional class improved in the three groups of patients. CONCLUSION: Percutaneous balloon mitral valvuloplasty is an effective, efficient method for the treatment of rheumatic mitral stenosis during pregnancy, after organogenesis, or at any time in a woman's life, as long as it is indicated according to clinical and echocardiographic evaluation criteria. PMID- 11412749 TI - [Troponine T as possible myocardial injury marker. Its application in myocardial stunning and silent ischemia]. AB - INTRODUCTION AND OBJECTIVES: The need for more specific, more sensitive and earlier biochemical markers of acute myocardial infarction, has led to the development of alternative methods to CK-MB). The aim of this work is to assess the usefulness of TnT measurement, in comparison with other markers for detecting transitory ischemic processes without necrosis in some experimental models. METHODS: The plasma levels of Troponine T, CK, CK-MB and adenosine were assessed as markers of ischemic myocardial injury. Two protocols were used: in Series I and II very brief (2 min ischemia with 3-min reperfusion) repeated (20 episodes) ischemias were induced, while Series III involved a single 15-min ischemia with a 60-min reperfusion. In Series I the coronary occlusor was placed close to the anterior descending coronary artery (AD); in Series II and III it was placed distally in the AD. Blood samples were taken from the peripheral vein (PVB) and corresponding coronary segment vein; in a basal situation, during ischemia, upon reperfusion, after 24 hours, and after 5 and 10 days. The plasma levels of adenosine, troponine T, CK and CK-MB as well as general and regional function parameters were measured. RESULTS: In Series I we observed hypokinesis that lasted 10 days, reaching its maximum on days 4-5. In Series II and III regional function was restored by 24 hours. CK and CK-MB showed similar behaviour; they rose significantly when the chest was opened (p < 0.05) reaching the highest value at 24 hours in all the series. Adenosine rose significantly only during reperfusion (p < 0.05). Troponine T increased after ischemia but not before, remained high for 5 days in all series (PVB). CONCLUSIONS: Troponine T rises in absence of necrosis, preferably when the ischemia is longer. PMID- 11412750 TI - [Regional diastolic function is more preserved in viable than non-viable myocardium. Demonstration by pulsed-wave Doppler tissue imaging in basal conditions]. AB - INTRODUCTION AND OBJECTIVES: The aim of this study was to analyze the differences in regional diastolic function between viable and non-viable myocardium when assessed by pulsed-wave Doppler tissue imaging performed in basal conditions. PATIENTS AND METHODS: The study population included 21 patients with three-vessel disease and regional systolic dysfunction. These patients underwent transthoracic echocardiographic study and pulsed-wave Doppler tissue imaging in basal conditions and, in addition, stress echocardiography with dobutamine performed by a different investigator. RESULTS: Three-hundred and twenty-two segments were studied, 140 of which (43%) had systolic dysfunction. Of the 140 segments with systolic dysfunction, 52 (37%) were considered hypokinetic by transthoracic echocardiography, 80 (57%) akinetic and 8 (6%) dyskinetic. As assessed by dobutamine echocardiography, 67 segments (48%) were considered viable and 73 (52%) non-viable. Viable segments had a higher peak velocity of the early diastolic wave e (5.5 +/- 1.9 vs. 4.7 +/- 2.0 cm/s; p = 0.03). An e/a ratio < 1 was more frequent in non-viable versus non-viable segments (52 vs. 70%; p < 0.05). There were no differences in relation to regional isovolumetric relaxation time and peak velocity of a wave. Although peak velocity of s wave was lower in non-viable segments, differences were not statistically significant. CONCLUSION: Compared with non-viable segments, viable myocardial segments have less impaired regional diastolic function as assessed by pulsed-wave Doppler tissue imaging. PMID- 11412751 TI - [Clinical outcome and health related quality of life in patients undergoing coronary angioplasty with balloon or stent. A prospective multicenter study]. AB - INTRODUCTION AND OBJECTIVES: Clinical outcome and health related quality of life after coronary angioplasty with stent or balloon are insufficiently studied in routine practice. The aim of the present study was to assess the impact of angioplasty on the clinical results and quality of life in real clinical practice. PATIENTS AND METHODS: All the consecutive patients undergoing angioplasty with stent or balloon attending two Spanish tertiary hospitals from October, 1997 to July, 1998 were evaluated at baseline and one year after discharge from hospital with a structured clinical questionnaire and the generic SF-36 quality of life questionnaire. RESULTS: Three hundred ninety-seven patients with a mean age of 63 years were included in the study. A stent (or stent plus balloon) was implanted in 342 patients and angioplasty with a single balloon was performed in 55 patients. More advanced coronary disease and suboptimal lesions for treatment were found in patients treated with only balloon. At one year of follow up the total mortality was 6% and 54% of the patients were free of angina. Mean baseline scores of the SF-36 questionnaire were remarkably low (35 for physical health and 45 for mental health). At one year the scores achieved levels similar to those of the general Spanish population matched for age and sex (45 for physical health and 52 for mental health). Independent predictors of quality of life at one year were the following: baseline quality of life, age, sex, comorbidity, previous hospitalizations, hospital where the patient was attended, symptoms at admission and late angina. CONCLUSIONS: a) After percutaneous myocardial revascularization the mean quality of life achieved was similar to that of the general population; b) different clinical subgroups did not achieve these levels, although improvement was similar to that of the remaining subgroups PMID- 11412752 TI - [Outcomes of coronary artery surgery: determinants of quality of life related to postoperative health]. AB - BACKGROUND AND OBJECTIVES: Little is known of the clinical and nonclinical determinants of health related quality of life after coronary artery bypass graft in routine clinical practice. The aim of this study was to assess the quality of life and its determinants after a first coronary bypass in a representative population of Catalonia, Spain. PATIENTS AND METHOD: Clinical and quality of life questionnaires were given to all the patients (n = 710) undergoing a first coronary bypass in private and public Catalan hospitals, prior to surgery and at six months and one year of follow-up. Quality of life was assessed with the DASI and the SF-36. RESULTS: The rate of clinical events at one year was 23%. The mean quality of life improved to levels slightly below those in general population; with greater changes reported in physical than in mental condition although the latter was less impaired. In 24%, the quality of life scores at one year were below 1.5 standard deviations of those in the general population. Females, patients with comorbidity and those with public health care insurance showed lower quality of life scores. Independent predictors of one-year quality of life included initial quality of life scores, public insurance, comorbidity, gender, age and chronic disease. Postoperative angina and dyspnoea were also associated with quality of life. CONCLUSION: The mean quality of life improves after coronary bypass, although up to one fourth of the patients may have unsatisfactory one-year clinical or quality of life outcome. Female patients, public insurance and comorbidity predict a worse quality of life. PMID- 11412753 TI - [Guidelines of the Spanish society of cardiology on requirements and equipment of the coronary care unit]. AB - The conceptual change and the dynamic care of patients observed in coronary care units, in recent years are reported. The coronary care unit is increasingly conceived as an intensive care unit for patients with acute or chronic cardiopathies with severe complications. Criteria for rational planification, functional and hierarchic organization and basic medical care arguments are established. The coronary care unit can not be considered as an isolated facility, but should rather be integrated in the cardiology department, under the direction of a cardiologist. The authors point out the basic, physical structure and characteristics of the equipment in the coronary care unit and the intermediate cardiac care unit, as well as the newly created chest pain units. Finally, we comment on the human resources (medical, nursing and administrative) and the criteria for admission and discharge in the coronary care unit. PMID- 11412754 TI - [New pharmacological agents in heart failure]. AB - Heart failure is a common and growing public health problem, with increasing incidence and prevalence over the last 2 decades. Despite improvements in its current management, heart failure is still associated with significant morbidity and mortality. This has motivated the search for newer therapeutic modalities, which are based on a better understanding on the pathophysiologic events that lead to heart failure. This review summarizes the potential role of new pharmacological agents in the treatment of heart failure. These potential new agents can be classified according to their role in the modulation of the main pathophysiologic abnormalities that characterized heart failure, that include: cellular-extracellular abnormalities, endothelial dysfunction, neurohormonal and immunologic activation. PMID- 11412755 TI - [Current topics in vascular biology: an update for the 21st century]. AB - Although taken for granted in the present day, the central role played by the blood vessels themselves in regulating the cardiovascular system under physiologic and pathologic conditions has only been understood in the last several decades. The field of vascular biology grew out of this appreciation of the primacy of the vasculature in cardiovascular pathologies. In this review, we come up to date on several topics of particular interest to the field of vascular biology as it enters the 21st century: namely, the role of oxidative stress in vascular function and dysfunction; angiogenesis and angiogenic gene therapies; and emerging biomarkers of atherosclerosis. Each of these areas is likely to experience significant advances in the coming years that will directly impact clinical decision-making and care of patients with cardiovascular diseases. PMID- 11412757 TI - [Left paracardial thymoma]. PMID- 11412758 TI - [Concurrent long QT and Brugada syndrome in a single patient]. AB - We present a 61-year-old patient with previous cardiac arrest and frequent syncopal spells. ECG showed a typical Brugada pattern and a QTc interval of 425 ms. During programmed ventricular stimulation a self-limited syncopal polymorphic ventricular tachycardia was induced. On diagnosis of the Brugada syndrome an implantable cardioverter defibrillator was implanted. Two days later two episodes of polimorphic ventricular arrhythmia were converted by the device. The ECG at this time showed a prolonged QTc of 500 ms in addition to a typical Brugada pattern. Atenolol was started and after a 36-month follow-up the patient has remained asymptomatic without arrhythmic events. In conclusion, this patient has the Brugada syndrome and also fulfills the clinical and ECG characteristics of the Long QT syndrome. These findings suggest a genetic link between the two syndromes. PMID- 11412759 TI - [Syncope due to parapharyngeal space lesions syncope-syndrome]. AB - The clinic association between tumors in the neck and syncopal attacks has been well documented but uncommon. The pathogenesis of this type of syncope has been attributed to an specific sinus node disfunction and several diseases that affect pacemaker activity, cardiac output and blood supply to the brain are able to produce it. The most common reflex cardiovascular sindromes linked to the IX nerve are carotid sinus syndrome and glossopharyngeal neuralgia-asystole syndrome. Some patients with a pathological involvement of the parapharyngeal space also suffers from syncope but the clinical picture of these patients differs so greatly from the other two syndromes, which suggests specific aetiological, clinical and therapeutical features so the term "parapharyngeal space lesiones syncope-syndrome" has been proposed for this entity. We report a case of a patient with a neck tumor in the parapharyngeal space with syncope as the major symptom. PMID- 11412760 TI - [Recurrent syncope due to late paroxysmal atrioventricular block after Senning repair]. AB - A 14-year old male with d-transposition of the large arteries and Senning repair should have presented four syncopal episodes with spontaneous recovery the year prior to admission is reported. Following conventional diagnostic procedures, including electrophysiologic study, no abnormalities were detected. Therefore, an insertable Holter was implanted, and two months later the patient presented a syncopal episode secondary to 15 second paroxysmal atrioventricular block. A dual chamber pacemaker was implantated and nine months later the patient remains syncope-free. PMID- 11412761 TI - [Endomyocardial fibrosis. Presentation as a right ventricular tumor]. AB - Endomyocardial fibrosis is an uncommon disease in Spain, with the most frequent clinical feature being congestive heart failure syndrome. Complete atrioventricular block is exceptional. Two-dimensional echocardiography is a fundamental investigation achieving the diagnosis. Surgical treatment is recommended in symptomatic patients. We present a patient with heart failure secondary to endomyocardial fibrosis with complete atrioventricular block and an infrequent echocardiographic image of a polycystic mass in the free right ventricular wall. PMID- 11412762 TI - [Adenosine in the preexcitation syndrome]. PMID- 11412763 TI - [Upper gastrointestinal endoscopy and risk of endocarditis]. PMID- 11412770 TI - [New diagnostic criteria for myocardial infarction: order in chaos]. AB - An expert committee of the European Society of Cardiology and the American College of Cardiology recently redefined the criteria for the diagnosis of myocardial infarction. The new nomenclature is based on the use of new, biochemical markers of myocardial necrosis (troponin, CK-MB mass) which are more sensitive and specific than those previously used (CK, CK-MB activity). The new criteria adapts to the real possibilities in clinical practice and presents the inconvenient of differing from the established criteria used as epidemiologic, prognostic and therapeutic references. Nonetheless, since there had been different criteria for diagnosing myocardial infarction in the past, the new nomenclature will represent a common way of referring a diagnosis with important practical implications. PMID- 11412771 TI - [Concomitant surgery for atrial fibrillation in the patient undergoing mitral surgery]. PMID- 11412772 TI - [Coronary bypass surgery in the elderly]. PMID- 11412773 TI - [Unstable angina in octogenarians: is an invasive approach feasible and effective?]. AB - BACKGROUND AND OBJECTIVE: Octogenarian patients with unstable angina are usually managed more conservatively despite having a worse prognosis. Studies of balloon angioplasty in the elderly have demonstrated a higher incidence of adverse events but the new advances (mainly stenting) have improved the results. We evaluated the efficacy and safety of an invasive approach in octogenarians with unstable angina. PATIENTS AND METHOD: From January 1996 to October 1999, 100 patients at least 80 years old with unstable angina were admitted to our unit and among these, 74 (74%) underwent percutaneous revascularization. We evaluated immediate results, in-hospital events and clinical follow-up. RESULTS: A total of 145 lesions were treated in 74 patients. The stent implantation rate was 79%. The success rate was 92%. Two patients died during hospitalization due to cardiac causes and 1 patient had a non-Q infarction. At follow-up, 24 +/- 12 months (range: 4 -50 months) 14 patients died (19.4%). New revascularization was performed in 10 patients (13.5%), 9 with PTCA and 1 with surgery. The survival rate free of death and infarction in the first year was 89.2%. At the end of follow-up 58 patients were alive (78.4%), 45 asymptomatic and 13 had stable angina, class I or II. CONCLUSIONS: The results of stent implantation in octogenarians were good with a 92% procedural success. Ninety-six percent of patients were free of death and infarction during hospitalization and 78.4% of the patients remained alive, most of them asymptomatic at the end of follow-up. PMID- 11412774 TI - [Improvement of endothelial function in patients with hypercholesterolemia and normal coronary arteries with lipid-lowering therapy]. AB - INTRODUCTION AND AIMS: In patients with coronary risk factors the presence of endothelial dysfunction in epicardial arteries has been documented. The purpose of this study was to determine whether endothelial dysfunction, documented hypercholesterolemic patients and angiographically normal coronary arteries, improves by reduction and normalization of lipid levels. PATIENTS AND METHOD: In 10 patients with hypercholesterolemia and normal coronary angiography, the endothelium-dependent coronary vasomotion was studied by intracoronary infusion of acetylcholine into the left anterior descending coronary artery. Vasomotion changes in response to acetylcholine were analyzed by quantitative angiography. Five patients without coronary risk factors and normal coronary arteries formed the control group. Patients with hypercholesterolemia were treated with lipid lowering therapy (diet and lovastatin) and endothelial function was reevaluated after 24 +/- 4 months. RESULTS: In the initial study, hypercholesterolemic patients compared with the control group showed a vasoconstrictor response to serial doses of acetylcholine(10(-6) M, 10(-5) M, 10(-4)M) indicative of endothelial dysfunction (study group: -0.3 +/- 10%, -6 +/- 4%, -18 +/- 10% vs control group: -0.6 +/- 6%, -2 +/- 6%, 3+/-6%; p < 0.01 to 10(-4) M acetylcholine dose. During follow-up hypercholesterolemic patients who a significant reduction in total cholesterol levels and LDL. Compared to first study, at follow-up, there was an improvement in the response to acetylcholine (-0.4 +/- 4%, -3 +/- 6%, -3 +/- 10%; p<0.001 vs basal values at 10(-4) M acetylcholine concentration). Reduction in total cholesterol during follow-up was related to the improvement in the vasoconstrictor response to acetylcholine (r=0.53; p< 0.05). CONCLUSION: In patients with hypercholesterolemia and angiographycally normal coronary arteries with documented endothelial dysfunction, the reduction and normalization of lipid levels during follow-up may improve endothelium-dependent coronary vasomotion. PMID- 11412775 TI - [Macroscopic and microscopic study of the right coronary artery after radiofrequency catheter ablation of cavotricuspid isthmus in an experimental model]. AB - INTRODUCTION AND OBJECTIVES: The right coronary artery (RCA) is found in the AV groove, just below the ablation target of atrial flutter, the cavotricuspid isthmus (ICT). After radiofrequency (RF) ablation with standard catheters, there have been no reports of complications, but it may not be successful in 10% of the cases. However, the use of irrigated tip catheters, which create deeper lesions, might potentially damage the coronary tree. METHODS: We analyzed the effects of ICT RF ablation on the coronary tree, with macroscopic and microscopic anatomopathological study. Ablation on ICT was performed using an irrigated tip catheter in 16 pigs and was randomly compared with a standard ablation catheter. There were no clinical problems, modifications of ST or new arrhythmias during the ablation. The heart was extracted at 1 week. RESULTS: The macroscopic study demonstrated that the right coronary artery was preserved, along the ICT, surrounded by the epicardial fat. The microscopic study showed an inflammatory reaction in the epicardial fat that reached the adventitia of the coronary artery, but never produced necrosis in 14 cases. The muscular wall of the artery had signs of focal inflammation without endothelium involvement in 2 cases. However, we found a subepicardial vein completely damaged by the radiofrequency. CONCLUSIONS: Although the right coronary artery is located just below the isthmus, no necrosis was observed in any case. However, there was an inflammatory reaction that focally reached the muscular wall. The veins may be damaged in RF ablation, perhaps due to the different blood flow of each vessel. PMID- 11412776 TI - [Partial left atriectomy in the treatment of atrial fibrillation associated with mitral valve disease]. AB - BACKGROUND: Atrial fibrillation is frequently observed under conditions associated with atrial dilatation. Atrial size is a factor related to the genesis and maintenance of atrial fibrillation. Predictive parameters of persistence of atrial fibrillation after maze procedure are atrial size and long duration of atrial fibrillation. The aim of this study was to investigate the effects of surgical left atrial reduction in chronic atrial fibrillation by mitral valvulopathy. PATIENTS AND METHOD: nineteen patients with chronic atrial fibrillation and dilated left atrium undergoing mitral valve procedures were included in this prospective study: group I with left atrial reduction (10 patients) and group II including control (9 patients). Both groups were with similar preoperative characteristics. RESULTS: At the mean follow-up of 12 months, all the patients in group II had chronic atrial fibrillation, and 7 patients in group I showed in atrial rhythm (p < 0.003). The patients in whom atrial fibrillation continued after surgery showed left atrial area of 33.8 +/- 12.3 cm2 and a volume of 98.5 +/- 53.9 ml; and the patients with normal rhythm had a left atrial area of 24.5 +/- 5.3 cm2 and a volume 60.3 +/- 21.2 ml. CONCLUSIONS: Preliminary results indicate that surgical left atrial reduction in patients with chronic atrial fibrillation may be a mechanism implicated in the elimination of arrhythmia after surgery. PMID- 11412777 TI - [Coronary surgery in elderly patients. In-hospital and long-term results]. AB - BACKGROUND: Taking into account the steady increase in the number of elderly patients requiring coronary artery bypass grafting, we sought to analyze the in hospital and long-term evolution of a group of elderly patients (>/= 75 years) who underwent coronary artery bypass grafting, and to identify clinical predictors of mortality and long-term symptoms. METHODS: Between April 1996 and February 2000, 207 patients older than 75 years of age who had undergone coronary bypass grafting were prospectively and consecutively analyze. Mean age was 78.4 +/- 2.7. RESULTS: An average of 2.6 grafts/patients was constructed. Left mammary artery was used in 93% of patients. The in-hospital incidence of heart failure, atrial fibrillation, preoperative infarction and stroke was 38%, 29%, 4.8% and 2.8% respectively. The in-hospital mortality rate was 5.8%. Mean follow-up was 18 months (25th an 75th percentiles 9-29). Late mortality rate was 4.1% in eight patients. Excluding the in-hospital deaths, the estimated probability of survival (Kaplan-Meier) at 3 years was 94% and the survival freedom from symptoms was 86%. A multivariate analysis showed that only age was predictor of in-hospital mortality (OR 1.16, p = 0.009). Only peripheral vascular disease was found as a predictor of symptoms during the long-term follow-up (p = 0.001). CONCLUSIONS: In this series of senile patients who underwent coronary surgery, those of an older age (> 80 years) showed a higher risk of in-hospital mortality. The presence of peripheral vascular disease is useful in the prognosis assessment of the group. PMID- 11412778 TI - [Evidence based medical treatment of heart failure]. AB - INTRODUCTION AND OBJECTIVES: Recommendations for the treatment of heart failure were carried out by a systematic review of the available evidence of the different pharmacologic treatments. MATERIAL AND METHODS: The review focused on the treatment of chronic and systolic heart failure. All the studies published in english about the pharmacologic treatment of heart failure where identified. The evidence of every pharmacologic treatment was classified according to: a) efficacy variables (reduction of mortality and hospitalizations, improvement of functional class, ejection fraction and exercise tolerance), and b) the level of quality of the evidence according to an evaluation scale. The evidence was also reviewed for the comparisons and the combinations of the pharmacologic treatments, as well as for the toxicity and costs of treatments. RESULTS: The recommendations were defined according to the NYHA functional class and were classified in the A, B and C categories according to the level of quality of the available evidence. The evidence on mortality was considered the most important. First line drugs, the alternatives and other possible treatments were take into account. CONCLUSIONS: There is enough evidence based on information about some variables such as reduction of mortality or hospitalizations to carry out treatment recommendations in all stages of heart failure. This point out the interest ant the priority of used them in the evaluation and improvement of the results of heart failure. PMID- 11412779 TI - [Hypothyroidism and myocardial damage in cardiac donors]. AB - BACKGROUND AND OBJECTIVES: Brain death may induce myocardial contractile dysfunction and hypothyroidism. This myocardial dysfunction is a limiting factor when considering donor heart for transplantation. Myocardial damage has shown to be the mechanism in which contractile dysfunction is established in donors. On the other hand, hypothyroidism has been evoked as a major cause of cardiovascular disorders in brain dead subjects. It is not known whether any relationship exists between hypothyroidism and myocardial damage that appear in brain death. In this study myocardial damage and thyroid status are quantified in a donor population, and whether a correlation between hypothyroidism and myocardial damage exists is analyzed. PATIENTS AND METHOD: Serum samples from 45 cardiac donors were examined for cardiac troponins T and I as myocardial damage markers, as well as thyroid hormones. Correlations between cardiac troponins and thyroid hormones were assessed. RESULTS: One or both cardiac troponins were elevated in 19 donors (42.2%). Hypothyroidism was a very frequent finding, since it was present in 39 donors (86.7%). No correlation was observed between any of the myocardial damage markers and thriiodothyronine levels. CONCLUSIONS: Hypothyroidism and myocardial damage are common in brain dead donors. No association exists between both disorders. PMID- 11412780 TI - [Guidelines of the Spanish Society of Cardiology on requirements and equipment in hemodynamic and interventional cardiology]. AB - In the last 20 years the work performed in catheterization laboratories has changed greatly, and while also taking the diagnostic aspects also into account, interventional cardiology has acquired an important role. Work in the catheterisation laboratory has evolved from only diagnostic studies of cardiac anatomy and function, and evaluation of potential surgical candidates, to interventional procedures mainly based on catheters techniques. As new diagnostic and interventional procedures are now available, human and technical requirements of the catheterization laboratory have changed. The aim of this report is to make an update of the requirements needed to perform diagnostic and interventional procedures in the cardiac catheterization laboratory. PMID- 11412781 TI - [Diabetes mellitus, inflammation and coronary atherosclerosis: current and future perspectives]. AB - Type 2 diabetes mellitus is a condition associated with an increased risk of coronary artery disease. This condition is currently reaching epidemic proportions in the Western world. Epidemiological studies have shown that insulin resistance and the constellation of metabolic alterations associated with type 2 diabetes mellitus such as dyslipidaemia, systemic hypertension, obesity and hypercoagulability, have an effect on the premature onset and severity of atherosclerosis. Albeit direct, the link between insulin resistance and atherogenesis is rather complex. It is likely that its complexity relates to the interaction between genes that predispose to insulin resistance and genes that independently regulate lipid metabolism, coagulation processes and biological responses of the arterial wall. The rapid development of molecular biology in recent years has resulted in a better understanding of the immune and inflammatory mechanisms that underlie insulin resistance and atherosclerosis. For example, it is known that nuclear transcription factors such as nuclear factor kappa beta and peroxisome proliferator-activated receptor are involved in atherosclerosis. The former modulates gene expression which encodes pro inflammatory proteins vital for the development of the atheromatous plaque. In the presence of insulin resistance there are multiple activating factors that could explain the early onset and severity of atherosclerosis. Glitazones, the new oral antidiabetic drugs and agonists of peroxisome proliferator-activated receptor, have been shown to improve peripheral insulin sensitivity and to also delay atherosclerosis progression in experimental models. Their beneficial effects have been linked to their anti-inflammatory effect. PMID- 11412782 TI - [Genetically modified animal models in cardiovascular research]. AB - It is a basic tenet of molecular and clinical medicine that specific protein complements underlie cell and organ function. Since cellular and ultimately organ function depend upon the polypeptides that are present, it is not surprising that when function is altered changes in the protein pools occur. In the heart, numerous examples of contractile protein changes correlate with functional alterations, both during normal development and during the development of numerous pathologies. Similarly, different congenital heart diseases are characterized by certain shifts in the motor proteins. To understand these relationships, and to establish models in which the pathogenic processes can be studied longitudinally, it is necessary to direct the heart to stably synthesize, in the absence of other peliotropic changes, the candidate protein. Subsequently, one can determine if the protein's presence causes the effects directly or indirectly with the goal being to define potential therapeutic targets. By affecting the heart's protein complement in a defined manner, one has the means to establish both mechanism and the function of the different mutated proteins of protein isoforms. Gene targeting and transgenesis in the mouse provides a means to modify the mammalian genome and the cardiac motor protein complement. By directing expression of an engineered protein to the heart, one is now able to effectively remodel the cardiac protein profile and study the consequences of a single genetic manipulation at the molecular, biochemical, cytological and physiologic levels, both under normal and stress stimuli. PMID- 11412783 TI - [The forest does not let you see the tree]. PMID- 11412784 TI - [Contrast echocardiography in the evaluation of postinfarction intraventricular thrombus]. PMID- 11412785 TI - [Treatment of acute myocardial infarction with the x-sizer coronary thrombectomy device]. AB - The abundant thrombi present in infarct-related arteries may impair the angiographic result of primary angioplasty in acute myocardial infarction. We describe the utilization of the thrombus-removal device X-Sizer before coronary stenting in 4 patients with acute myocardial infarction treated with primary angioplasty. The procedure was successful in all the cases and the patients were discharged without complications. PMID- 11412786 TI - [Cardiac failure due to traumatic aortic valve rupture]. AB - Cardiac failure due to valvular dysfunction is frequent as a complication of acute myocardial infarction, endocarditis or penetrating thoracic trauma affecting the heart and large vessels. Less frequently it is associate with nonpenetrating chest trauma. We presented a 69-year-old male with acute left heart failure after nonpenetrating thoracic trauma and sternum fracture. Transesophageal echocardiogram confirmed severe aortic regurgitation. A bilateral tear in the right coronary and non-coronary cusp was found. The aortic valve was removed and replaced with a number 27 Carpentier Edwards prosthesis. The postoperative course was uneventful. PMID- 11412787 TI - [Angina pectoris in the heart transplant recipient: evidence of reinnervation]. AB - Heart transplantation involves the removal of a sick heart together with its innervation and replacement with a donor heart isolated from the control of the autonomous nervous system of the recipient, therefore being, functionally dennervated. Dennervation conditions several alterations in cardiac physiology, such as the inability to experience pain during myocardial ischemia, so that theoretically these patients cannot present angina pectoris. However, several reports have shown evidence of reinnervation with isolated cases of transplanted patients with angina pectoris having been reported. We describe the case of a transplanted patient who showed typical effort angina, vascular disease of the graft and data of sympathetic reinnervation demonstrated by cardiac gammagraphy with metayodo-benzilguanidina-I-123. PMID- 11412788 TI - [A case of Carney syndrome]. AB - The Carney complex is an autosomal dominant syndrome characterised by multiple neoplasies, including myxomas at various sites (cardiac, cutaneous, mammary), spotty pigmentation, endocrine overactivity (Cushing's syndrome, acromegalia), testicular tumours and schwannomas. This report describes the case of a 43-year old woman with an acute inferior myocardial infarction, probably due to coronary embolization from a large left atrial myxoma and who presented this association. A brief review of the Carney complex is provided after discussion of this rare case that required an screening of family members. PMID- 11412789 TI - [The Cholesterol 2000 document: is it useful for renal patients?]. PMID- 11412795 TI - [XXII National Congress of the Spanish Society of Nuclear Medicine. Ciudad Real, Spain, 13-16 June 2001. Abstracts]. PMID- 11412803 TI - Female genital schistosomiasis: facts and hypotheses. AB - In this paper we summarise the parasitological, clinical and epidemiological characteristics of female genital schistosomiasis (FGS), a frequent manifestation of the infection with Schistosoma haematobium. Means to diagnose and treat lesions in the lower and upper genital tract are discussed. Based on clinical findings and available pathophysiological as well as immunological data it is conceivable that FGS of the cervix and vagina not only facilitates the infection with agents of sexually transmitted diseases, but presumably also alters the natural history of such infections. Two infectious agents are of particular concern: the Human Immunodeficiency Virus and the oncogenic Human Papilloma Viruses. Possible interactions and their consequences are discussed and research areas which should be addressed are outlined. PMID- 11412804 TI - A cDNA encoding Tc-MUC-5, a mucin from Toxocara canis larvae identified by expression screening. AB - Toxocara canis is an ascarid nematode parasite of canids. Larvae infect a wide range of accidental hosts including humans, in whom they are the aetiologic agent of visceral and ocular Larva migrans. The labile surface coat of T. canis larvae consists of a family of mucin glycoproteins termed TES-120, for which the cDNAs have recently been cloned. In this paper, we describe the identification of a novel cDNA (Tc-muc-5) encoding an apomucin by expression screening of a cDNA library with antiserum raised to T. canis excretory/secretory products, and compare the predicted Tc-MUC-5 protein with those of other T. canis mucins (Tc MUC-1-Tc-MUC-4) that include the TES-120 surface coat glycoproteins. Tc-MUC-5 has both a larger open reading frame and a more divergent sequence than the other T. canis mucins. It contains a putative signal peptide followed by two six-cysteine (SXC) domains, an extended threonine-rich central mucin core domain and two C terminal SXC domains. Amino acid composition analysis of secreted TES-120 glycoproteins revealed a distinct lack of lysine residues; while this finding is in agreement with the primary sequences of Tc-MUC-1-Tc-MUC-4, Tc-MUC-5 is conspicuous by its relative abundance of lysines (6.7%), suggesting that this protein is not part of the TES-120 family of surface coat proteins. PMID- 11412805 TI - Epidemiological surveillance of human hydatidosis by means of ultrasonography: its contribution to the evaluation of control programs. AB - Epidemiological surveillance for hydatidosis is based on initial determination and follow-up of the infestation rate in man, sheep and dogs. The use of ultrasonography (US) as a screening method has demonstrated its usefulness in detecting asymptomatic human carriers of abdominal hydatidosis. To evaluate the contribution of US to epidemiological surveillance within the framework of disease control programs, we performed 719 US studies in school children from 7 to 13 years of age, in rural areas of Rio Negro, Argentina, subjected to canine deparasitization during 1984/1986 and 1997/1998. In the first period, 15/268 (5.6%), while in the second, 5/451 (1.1%) carriers were detected (P < 0.0004). The average diameter of the cysts was 4.71 cm in 1984/1986 and 2.14 in 1997/1998. US as a mass screening method allows evaluation of early changes in human prevalence rates, closely related to infestation rates in sheeps and dogs, thus providing a sensitive indicator of the evaluation of control programs. PMID- 11412806 TI - Sensitivity of the polymerase chain reaction for the diagnosis of cutaneous leishmaniasis due to Leishmania (Viannia) guyanensis. AB - The sensitivity of the polymerase chain reaction (PCR) in 35 consecutive outpatients with cutaneous leishmaniasis caused by Leishmania (Viannia) guyanensis was evaluated using, as gold standard, the in vitro isolation of the parasite through culture of aspirates of the cutaneous ulcers. All isolates were identified using electrophoretic enzyme analysis. Patients were mainly young males with recent onset disease without prior specific treatment. PCR was performed using DNA extracted from fresh frozen biopsies of cutaneous ulcers. The reaction used a pair of oligonucleotides that amplify the conserved region of the minicircle molecule. PCR showed 100% sensitivity (95% CI from 90.0 to 100.0). These results were similar to the visualization of amastigotes in imprint preparations of cutaneous biopsy tissue and the inoculation of biopsy material in golden hamsters. Despite the high sensitivity of the PCR, in this particular clinical setting of cutaneous leishmaniasis caused by L. (V.) guyanensis in the Brazilian Amazon, it appears that the method of choice for diagnosis should be the direct visualization of amastigotes using imprint preparations and the PCR reserved for those patients with negative imprint results. PMID- 11412807 TI - Cardiovascular risk factors in Tanzania: a revisit. AB - In this assessment of cardiovascular risk factors, we examined the prevalence of selected risk factors according to the World Health Organisation (WHO) CARDIAC Study protocol and compared them with a similar study conducted more than a decade ago. The survey was carried out in Dar es Salaam (D, urban), Handeni (H, rural) and Monduli (Mo, semi-nomadic area). Subjects aged 47-57 were recruited randomly for blood pressure and anthropometrical measurements, 24 h urine collection and blood sampling. A structured questionnaire was used to obtain dietary information. The 1998 survey studied 446 subjects, while the 1987 survey included 496 men and women. The measured weight, body mass index (BMI) and prevalence of obesity (BMI > or = 30 kg/m(2)) increased significantly among women in the 1998 survey in rural Handeni and urban Dar. The overall prevalence of obesity was higher for women in the most recent survey (22.8%, P < 0.0001). Diastolic blood pressure (DBP) was higher in the most recent survey for women in Handeni. The overall prevalence of hypertension (blood pressure > 160/95 mmHg, or antihypertensive drug use), rose to 41.1% in 1998, (P < 0.001) for men and to 38.7% (P < 0.05) for women. The mean total serum cholesterol and prevalence of hypercholesterolaemia increased significantly in the most recent survey in the three studied areas. The overall prevalence of hypercholestrolaemia (serum cholesterol > 5.2 mmol/l) was higher in the 1998 survey for both men (21.8%, P < 0.0001) and women (54.0%, P < 0.0001). The mean HDL cholesterol increased significantly in the most recent survey, with a significant reduction in the mean atherogenic index, though these were still at higher levels (men 5.8, P < 0.0001; women 5.1, P < 0.0001 vs. 1987). A strong positive correlation was observed between blood pressure (SBP and DBP) and body mass index, total serum cholesterol and sodium to potassium ratio. These data suggest that for the past decade there has been an increase in the mean levels and prevalence of selected cardiovascular risk factors in Tanzania. PMID- 11412808 TI - Bancroftian filariasis in residents of Oman. PMID- 11412809 TI - A rapid and simplified method of DNA extraction for the detection of Brugia malayi infection in mosquitoes by PCR assay. AB - Currently used protocols for the extraction of filarial parasite DNA from mosquito samples are tedious and involve extensive use of expensive and hazardous chemicals. Therefore, in order to arrive at a simple procedure, four different methods (A, B, C and D) were tried for the extraction of DNA from mosquitoes infected with filarial parasite, Brugia malayi. Method D was found to be as efficient as the current procedure for the extraction of DNA from a single microfilaria in pools of 25 mosquitoes and the DNA was suitable for polymerase chain reaction (PCR) amplification, yielding a band of 322 base pairs with primers specific for B. malayi. Method D involved drying and crushing the mosquitoes to a powder, which was homogenized in 100 microl TE buffer, vortexed, boiled for 10 min, centrifuged at 14000 r.p.m. for 10 min, and the supernatant used for the PCR assay. Dot-blot hybridization confirmed the specificity of the PCR amplified fragment. The DNA extracted by this method was stable for about 1 year. When comparing with the standard method, the cost of a single PCR reaction, inclusive of DNA extraction, was reduced by 50% and the hands on time was minimized fivefold. Hence, this simple TE-based method is rapid, safe and also cost-effective in assessing the B. malayi infection in pools of vector mosquitoes. PMID- 11412810 TI - The Albert Dock Hospital, London: the original site (in 1899) of Tropical Medicine as a new discipline. AB - The London School of Tropical Medicine, and indeed the 'new' discipline of tropical medicine, originated in 1899 at the Albert Dock Hospital - situated in London's east-end. The founder of the discipline was Patrick Manson - with a great deal of political assistance from the British Secretary of State for the Colonies (Joseph Chamberlain). The hospital (originally opened in 1890) was rebuilt in 1937-1938 and ultimately demolished in 1993. PMID- 11412811 TI - Pharmacogenomics: will it change the field of medicine? PMID- 11412812 TI - Genetic mechanisms for hypersensitivity and resistance to the anticoagulant Warfarin. AB - Warfarin is the therapeutic of choice for maintenance anticoagualtion therapy. A principle caveat of this medication is that the dosage required to achieve the desired therapeutic effect varies up to 120-fold between individuals. Currently, there are no reliable means of prospectively identifying which patients will require either unusually high or low dosages. This dilemma puts patients at risk of therapeutic failure or potentially life-threatening overdosage during a prolonged trial-and-error period of establishing an individualized medication strategy. Pharmacogenetic research has revealed that extreme differences in the drug dose required to achieve the desired therapeutic response can be attributed to genetic variation in the genes encoding drug metabolizing enzymes, and cellular receptor proteins. The anticoagulant Warfarin represents a model system where there is evidence to suggest that both pharmacokinetic and pharmacodynamic mechanisms contribute to the overall variability in patient response. Here the current understanding concerning the influence of genetic variation in Warfarin pharmacokinetics is reviewed and the potential for similar genetic mechanism impacting on the pharmacodynamic response in man is explored. Diagnostic testing to identify subjects requiring low-dose Warfarin therapy is discussed in light of potential confounding or coexisting resistance to the drug effects. PMID- 11412813 TI - Automated detection of single nucleotide polymorphism in beta-2 adrenergic receptor gene using LCx(R). AB - Beta-2 adrenergic receptor (B2AR) agonists are the most widely prescribed rescue agents used in the treatment of asthma. Recent studies have indicated a relationship between a polymorphism at codon 16 of the B2AR gene, and the response to recurrent beta-agonist therapy. The B2AR polymorphism of interest involves a single nucleotide change from A to G, resulting in an amino acid change from Arginine (Arg) to Glycine (Gly). Clinical efforts to further investigate this relationship require an accurate, reliable and inexpensive method for detecting the polymorphism. In this study, we report an LCx(R) assay for the detection of a single nucleotide polymorphism at codon 16 of the beta-2 adrenergic receptor. This assay is capable of detecting patients harboring any of the three possible genotypes at this locus, namely, homozygous wild type, homozygous variant or heterozygous individuals with a single genomic DNA sample of 25-500 ng. It requires minimum hands-on time with automated detection. The assay would be suitable for use in research labs for screening of a large number of samples. We believe that this type of assay will facilitate research and clinical investigations in elucidating the association of SNPs with disease states, diagnosis, prognosis and treatment. PMID- 11412814 TI - Genotyping of cytochrome P450 2D6*3 and *4 mutations using conventional PCR. AB - Cytochrome P450 (CYP450) mixed-function mono-oxygenases, consisting of more than 30 enzymes, are responsible for the metabolism of a large number of drugs and metabolites. With the rapid advances in the human genome project, the role of genetic polymorphism in drug metabolism may become an important adjunct for rational drug therapy, and for the explanation of drug toxicity and interactions. This preliminary study modified a previously described procedure for genotyping CYP2D6*3 and *4. An additional step included uracil-DNA glycosylase for the prevention of "carry-over" contamination. DNA was extracted from peripheral blood using PureGene DNA Isolation kit. CYP2D6*3 and *4 sequences were amplified by PCR, followed by digestion with restriction endonuclease Msp1 and Mva1, respectively. Resulting fragments were analyzed by electrophoresis and visualized by ethidium bromide staining. Poor metabolizers of *3 mutation showed 168-, 82- and 20-bp bands, while those of *4 showed a single 355-bp band. Using these protocols, 22 individuals were genotyped, showing the following prevalence for *3 and *4: 0 and 3, respectively-comparable to those of the general population. This method provides a reliable means of genotyping CYP2D6*3 and *4. PMID- 11412815 TI - Pharmacogenetics: a new diagnostic tool in the management of antidepressive drug therapy. AB - Pharmacogenetics studies the genetic basis of an individual's ability to respond to pharmacotherapy. Variability of this response is a major problem. Fatal adverse drug reactions have been reported to be the fourth leading cause of death in the US. In depression, 30-40% of all patients do not respond sufficiently to the initial treatment and it can take up to 6 weeks for them to be identified. Much knowledge has been gathered throughout the last 3 decades about the genetic basis of pharmacokinetic variability. Genetic tests suitable for the routine laboratory are now available for some important metabolizing enzymes (e.g., CYP2D6, CYP2C19) identifying those individuals who are slow or fast metabolizers of certain drugs, many of which are widely used in the treatment of depression (e.g., tricyclic antidepressants). The possible use of these tests in the clinical practice of monitoring antidepressant therapy is discussed in relation to older phenotyping methods and therapeutic drug monitoring.Less well studied than the genetics of pharmacokinetics is the genetic basis of pharmacodynamic variability. As selective serotonin reuptake inhibitors (SSRI) have a wide therapeutic index, pharmacokinetic variability usually does not explain insufficient response to therapy. Recently, some excitement was caused by reports on serotonin transporter gene polymorphisms and their influence on the response to antidepressive therapy with SSRIs as this could provide an interesting diagnostic tool in assessing the chances of response to the most popular group of antidepressants at present. Current knowledge in this young field of research is summarized. PMID- 11412816 TI - Pharmacogenomics: implications for laboratory medicine. AB - Pharmacogenomics deals with the interactions of individual genetic constitution with drug therapy. It has potentially far reaching consequences for drug development and future treatment strategies, but also for clinical in vitro diagnostics. With increasing knowledge about interactions between genes and drug treatment, there will be an equally increasing demand for rapid and reliable diagnostic tests prior to the institution of therapy. In fact, it is very likely that pharmacogenetic tests will make up a significant proportion of total molecular biology testing in the coming years. Therefore, this review focuses on the implications of pharmacogenomics on the clinical laboratory. PMID- 11412817 TI - Development of an amperometric biosensor based on glutathione peroxidase immobilized in a carbodiimide matrix for the analysis of reduced glutathione from serum. AB - The development of an amperometric biosensor for the reduced glutathione determination in serum is described. The biosensor is based on glutathione peroxidase (GSH-Px, EC 1.11.1.9) immobilized onto a pyrolytic graphite-working electrode using carbodiimide as enzymatic condensing reagent. This resulted in an amperometric biosensor with good sensitivity and stability. The reduced glutathione (GSH) was enzymatically converted to glutathione disulfide (GSSG) in the presence of hydroperoxide, which was monitored amperometrically by its electrooxidation at +0.65 V vs. SCE (saturated calomel electrode). Glutathione measurement was carried out by maintaining the ratio between GSH and hydrogen peroxide at 2:1 (25 degrees C). The amperometric response of the biosensor was linearly proportional to the GSH concentration between 1.9x10(-5) and 1.4x10(-4) mol/l, in 0.1 mol/l phosphate buffer (pH=7.8), containing 0.1 mol/l KCl and 0.5 mmol/l Na(2)H(2)EDTA, as the supporting electrolyte. In presence of interfering compounds, the recoveries ranged between 97.2% and 110.7%. The biosensor useful lifetime was at least 2 months when it was evaluated after continuous use. Serum samples analyzed by this biosensor showed a good correlation with the results from the spectrophotometric method (Ellman's reagent) used as reference, presenting relative deviations lower than 7.0%. The low apparent Michaelis-Menten constant value, K(M)(app)=1.6 mmol/l, demonstrated that GSH-Px immobilized on pyrolytic graphite exhibited a high affinity to GSH, without loss of enzymatic activity. PMID- 11412818 TI - Continuous monitoring of arylesterase in human serum. AB - Arylesterase (EC 3.1.1.2) activity in serum was specifically measured using thiophenyl acetate in a mechanized assay at 37 degrees C with 4 bromophenylboronic acid as inhibitor of cholinesterase and hexacyanoferrate-III as indicator. The systematic development of a routine method, apparent limitations of thiophenyl compounds as arylesterase substrates, some kinetic constants of the enzyme, analytical variables such as precision (within-run <2% and between-run <2.5% relative standard deviation) and a preliminary reference interval (19.5-52.4 kU/l) for adults are described. PMID- 11412819 TI - Disease pattern recognition testing for rheumatoid arthritis using infrared spectra of human serum. AB - BACKGROUND: In view of the importance of the diagnosis of rheumatoid arthritis, a novel diagnostic method based on spectroscopic pattern recognition in combination with laboratory parameters such as the rheumatoid factor is described in the paper. Results of a diagnostic study of rheumatoid arthritis employing this method are presented. METHOD: The method uses classification of infrared (IR) spectra of serum samples by means of discriminant analysis. The spectroscopic pattern yielding the highest discriminatory power is found through a complex optimization procedure. In the study, IR spectra of 384 serum samples have been analyzed in this fashion with the objective of differentiating between rheumatoid arthritis and healthy subjects. In addition, the method integrates results from the classification with levels of the rheumatoid factor in the sample by optimized classifier weighting, in order to enhance classification accuracy, i.e. sensitivity and specificity. RESULTS: In independent validation, sensitivity and specificity of 84% and 88%, respectively, have been obtained purely on the basis of spectra classification employing a classifier designed specifically to provide robustness. Sensitivity and specificity are improved by 1% and 6%, respectively, upon inclusion of rheumatoid factor levels. Results for less robust methods are also presented and compared to the above numbers. CONCLUSION: The discrimination between RA and healthy by means of the pattern recognition approach presented here is feasible for IR spectra of serum samples. The method is sufficiently robust to be used in a clinical setting. A particular advantage of the method is its potential use in RA diagnosis at early stages of the disease. PMID- 11412820 TI - Comparison of thyroid function in pregnant and non-pregnant Asian and western Caucasian women. AB - BACKGROUND: Gestational thyrotoxicosis may be more prevalent in Asian women. METHODS: We have measured thyroid function, ferritin and bone specific alkaline phosphatase (BALP) as peripheral markers of thyroid function and hCG in Asian and western Caucasian women in non-pregnant and early pregnancy. RESULTS: TSH was lower in Asian women in non-pregnancy but not during normal pregnancy and this may reflect increased sensitivity of the thyroid gland to thyroid stimulation in the Asian population. No ethnic difference was found in FT3, FT4 or hCG but ferritin was lower and BALP higher in Asian women whether pregnant or not and this may be a reflection of iron balance and vitamin D status. CONCLUSIONS: We found during normal pregnancy that dynamic patterns of change for thyroid hormones and hCG are not different in Asian and western Caucasian women. We have developed gestation related reference intervals, which are a pre-requisite to the study of ethnic differences in gestation thyrotoxicosis in pregnancy. PMID- 11412821 TI - A new homogeneous enzyme immunoassay for thyroxine using glycogen phosphorylase b thyroxine conjugates. AB - BACKGROUND: [corrected] Measurement of serum thyroxine (T(4)) concentration is important for diagnosis of thyroid gland diseases. We developed a practical homogeneous enzyme immunoassay for thyroxine analysis in unextracted sera. METHODS: A thyroxine derivative conjugated to a reactive sulfhydryl group of glycogen phosphorylase b (GPb). Conjugation caused inhibition of enzyme activity and the enzyme conjugate was re-activated upon the binding of a polyclonal anti T(4) antibody. Antibody-activation was blocked by the presence of free T(4). RESULTS: Conjugation affected the allosteric character of the enzyme and the K(m) for the allosteric activator AMP was increased 28 times, while anti-T(4) antibody partially reversed this effect. The optimum concentration ratio of enzyme conjugate to anti-T(4) antibody was determined, and T(4) was measured with desired sensitivity and accuracy in the range between 10 and 240 microg/l. Furosemide was used to inhibit the interaction of thyroxine with serum T(4) binding sites. Human serum T(4) values obtained by this method correlated well with those obtained by a radioimmunoassay (y=1.9+1.0x, r=0.97, N=72). CONCLUSIONS: Chemical modification of glycogen phosphorylase b with a T(4) derivative led to the development of a simple homogenous enzyme immunoassay for T(4) analysis with the desired sensitivity and accuracy. PMID- 11412822 TI - Cartilage and bone biological markers in the synovial fluid of osteoarthritic patients after hyaluronan injections in the knee. AB - OBJECTIVE: To evaluate synovial fluid levels of cartilage and bone biological markers after repetitive intra-articular injections of sodium hyaluronate (HA) in knee osteoarthritis (OA) patients. METHODS: Twenty patients with knee OA were evaluated before and after 5 weekly injections of HA. To study cartilage and bone biological markers, synovial fluid and urine samples were collected simultaneously with the first (FI=week 0) and before the last injection (LI=week 4) of HA. Not commercially available markers (cartilage oligomeric matrix protein (COMP), proteoglycan monomers and cyanogen bromide peptide 11 of the type II collagen chains (alpha (II) CA11B)) were determined by an indirect inhibition ELISA developed and standardized in our laboratory. RESULTS: We found a significant reduction in levels of proteoglycan monomers (30+/-16 vs. 22+/-10 microg/ml, p<0.05), an increase in COMP concentration (2.9+/-0.9 vs. 3.6+/-0.9 microg/ml, p<0.05) and osteocalcin (BGP) levels (8.7+/-8 vs. 11.9+/-9 ng/ml, p<0.05). No significant changes were observed in the levels of alpha (II)CB11B), metalloproteinase-1 (MMP-1) or pyridinium cross-link/creatinine (Pyr/Cr). CONCLUSIONS: HA could elicit an indirect response on the cartilage and bone metabolism due to the increased overuse of the joint caused by the analgesic effect of HA. However, a direct HA action on the metabolism of chondrocytes must not be ruled out. PMID- 11412823 TI - Matrix interference in serum total thyroxin (T4) time-resolved fluorescence immunoassay (TRFIA) and its elimination with the use of streptavidin-biotin separation technique. AB - In our development of total serum thyroxin TRFIA using an immobilized second antibody (S-Ab) as the separation agent, we observed a significant measurement bias caused by a matrix interference when the immobilized S-Ab had a relatively low binding capacity for the primary anti-T4 monoclonal antibody (McAb). Therefore, we employed a new separation system based on the highly active surface streptavidin and biotinylated anti-T4 McAb. Our results indicate that the matrix interference was removed and the assay performance was improved with the use of streptavidin-biotin separation technique. In our method, microwells were first coated with biotinylated BSA and then a streptavidin solution in the presence of 1% BSA was added to allow streptavidin to be immobilized via the pre-coated BSA biotin. Surface streptavidin prepared in this protocol expressed a significantly increased binding capacity for the biotinylated anti-T4 McAb, compared to the passively adsorbed S-Ab for binding the original anti-T4 McAb. The immunoreactions between the biotinylated anti-T4 McAb and the T4 in the standard or sample or the europium-labeled T4-BSA conjugate mainly occurred in liquid solution, and then the immune complex was specifically trapped by the surface streptavidin and isolated from the free trace by washing. Serum TT4 TRFIA based on surface streptavidin was accurate, precise and economic, maintained all the merits of the directly immobilized surface antibodies. PMID- 11412824 TI - Chromosomal instability detected by fluorescence in situ hybridization in Japanese breast cancer patients. AB - The relationship between chromosomal instability (CIN) and prognostic factors was investigated in 31 breast cancers and 5 benign breast lesions (three fibroadenomas and two papillomas). Using fluorescence in situ hybridization (FISH) with chromosome-specific DNA probes of chromosomes 1, 2, 6, 7, 10, 11, 17 and 18, CIN for each case was determined. CIN varied from 8.1% to 59.3% among the breast cancer patients tested, and was significantly higher than that observed in the benign breast lesions (p<0.01). Moreover, CIN showed a significant correlation with lymph node metastases (p<0.05) and estrogen receptor negativity (p<0.01). These findings suggest that CIN might be useful in the prediction of the biological aggressiveness of breast cancers. PMID- 11412825 TI - Pravastatin therapy increases procollagen I N-terminal propeptide (PINP), a marker of bone formation in post-menopausal women. AB - BACKGROUND: The aim of our study was to evaluate whether pravastatin treatment affected biochemical markers of bone turnover. METHODS: Thirty-six hypercholesterolemic post-menopausal women, not on hormonal replacement therapy, were selected from a population study evaluating factors affecting cholesterol response to pravastatin. After a 6-week period on a 30% fat diet, participants received treatment with 20 mg/day of pravastatin during a 16-week follow-up period. Pre- and post-treatment samples were analyzed for procollagen I aminoterminal peptide (PINP) and bone alkaline phosphatase (bAP) as markers of bone formation, carboxyterminal telopeptide of collagen I (CTX) as a marker of bone resorption, and procollagen III aminoterminal propeptide (PIIINP) as a marker of fibrogenesis. RESULTS: Total cholesterol decreased from 7.26+/-0.83 to 6.1+/-0.77 mmol/l with pravastatin treatment. PINP levels significantly increased (from 33.6+/-13 to 37.4+/-16, p=0.03) without changes in bAP or CTX. Individual changes in PINP correlated with individual reduction in cholesterol levels (r=0.337, p=0.04). There was no significant change in PIIINP concentration. CONCLUSIONS: Pravastatin treatment increased PINP levels, a marker of bone formation, in hypercholesterolemic, post-menopausal women, without affecting bone resorption. PIIINP concentration, a marker of liver fibrogenesis, was not affected by the treatment. PMID- 11412826 TI - Severe hypertriglyceridemia with plasma inhibitory factor(s) on lipoprotein lipase activity in a patient with a common Ser(447)-Ter LPL mutation. AB - Severe hypertriglyceridemia is a major risk for acute pancreatitis. So far, several mutations on the lipoprotein lipase (LPL) gene causing type I hyperlipidemia have been identified. However, the common mutation Ser(447)-Ter has been recently proposed to have a lowering effect on serum triglyceride concentrations in the general population. In this study, we analyzed blood from a patient suffering from severe hypertriglyceridemia and pancreatitis with the mutation on the lipoprotein lipase gene, Ser(447)-Ter. The patient's plasma showed inhibitory effects on the LPL activities from normal subjects. The bottom fraction separated by ultracentrifugation revealed enhanced effects as an inhibitory factor. The inhibitory effect observed in the bottom fraction was dose dependent, stable at treatment of 65 degrees C for 30 min, and decreased significantly after being dialyzed using membranes with a cut-off molecular weight of 3500 or 6000 Da. The inhibitory effect was significantly higher when the post-heparin plasma was used from the patient or a subject with the same LPL mutation as an LPL source, compared to that from normal subjects. These results suggest that the patient has inhibitory factors in his plasma. Such inhibitory factors might cause severe hypertriglyceridemia in a case with the common mutation, which has been proposed to show the lowing effect on serum triglyceride concentrations in the general population. PMID- 11412827 TI - CBQCA assay of primary amine losses during hemodialysis. AB - Nitrogen loss during hemodialysis is an important issue since all steps taken to improve clearance of urea will also increase loss of amino acids and other normally recyclable sources of nitrogen. Most dialyzers exclude albumin and other proteins, which react well with dyes commonly employed to measure protein, but do allow passage of mono- and oligopeptides that go undetected in protein assays. The CBQCA reagent provides a highly sensitive assay of primary amines that will detect amino acids, low molecular weight polypeptides, and high molecular weight proteins. We collected dialysate using a split stream technique from 28 chronic dialysis patients. With bovine albumin as the standard, the CBQCA assay reported 24.3+/-9.8 g albumin-equivalents (mean+/-S.D.) per dialysate whereas a Coomassie blue assay measured 1.19+/-0.78 g. The CBQCA assay values were substantially higher than previously reported amino acid losses. The CBQCA fluorescent assay for amines provides a simple assay for quantifying primary amine losses in dialysate fluid. PMID- 11412828 TI - Lipid peroxidation, osmotic fragility and antioxidant status in children with acute post-streptococcal glomerulonephritis. AB - Plasma and erythrocyte samples from acute post-streptococcal glomerulonephritis (APSGN) children and control children were enrolled in this study. Lipid peroxidation (LPO), measured in terms of thiobarbituric acid-reactive substances (TBARS) was found to be significantly increased in plasma and RBCs of APSGN children (P<0.05) than in control children. Osmotic fragility of erythrocytes was examined. RBCs of APSGN patients were found to be osmotically more sensitive towards hypotonic saline (50% hemolysis at 7 g/l saline) when compared to control RBCs (50% hemolysis at 4 g/l saline). The activities of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione S-transferase (GST) were significantly lowered (P<0.05) in APSGN RBCs when compared to control RBCs. Plasma ascorbic acid, reduced glutathione (GSH), RBC ascorbic acid, GSH and RBC total sulphydryl content (TSH) were significantly depleted in APSGN children relative to controls. The susceptibility of RBCs of APSGN children to lipid peroxidation was confirmed in this study. PMID- 11412829 TI - Highly sensitive and rapid method for determination of fluoride ion concentrations in serum and urine using flow injection analysis with a fluoride ion-selective electrode. AB - An apparatus for flow injection analysis (FIA) was developed to measure very low fluoride ion concentrations (<1 micromol/l). The analytical conditions of the apparatus were investigated, and the instrument was used to determine fluoride ion concentrations in serum and urine. All interferences caused by serum and urine matrices were eliminated using the proposed method. The recovery was almost 100.0% for serum and urine samples. The precision was within 4%. The results of determination of fluoride ion concentrations in the NIST Standard Reference Material of urine, SRM 2671a, agreed with the certified values. The detection limits in serum and urine were 0.016 and 0.16 micromol/l, respectively. The assay throughput was 15 samples/h in serum and 24 samples/h in urine. The mean fluoride ion concentrations in serum and urine samples from 53 young Japanese women were 0.383+/-0.158 micromol/l and 0.207+/-0.103 mg/g Cr, respectively. There was a significant correlation (r=0.39, p<0.01) between serum and urine fluoride ion concentrations. PMID- 11412830 TI - Analysis of guanidinoacetate and creatine by isotope dilution electrospray tandem mass spectrometry. AB - Guanidinoacetate methyltransferase (GAMT) deficiency is a disorder of creatine metabolism characterized by low plasma creatine concentrations in combination with elevated guanidinoacetate (GAA) concentrations. Although rare, GAMT deficiency has been identified in children with seizures, extrapyramidal movements, developmental delay, myopathies and behavioral abnormalities. Treatment with creatine monohydrate has been proven to be effective. We describe an isotope dilution electrospray tandem mass spectrometry (ES-MS/MS) assay for the simultaneous determination of plasma GAA and creatine using multiple reaction monitoring (MRM), d(3)-creatine as the internal standard and derivatization of GAA and creatine as butyl-esters. We analysed plasma of 16 healthy adults and 20 healthy children as well as three affected children. Plasma GAA concentrations were 5.02+/-1.84 micromol/l (mean+/-S.D.) in adults, 3.91+/-0.76 micromol/l in children age 5-10 years and 11.57, 15.16, 14.36 micromol/l in children with GAMT deficiency. Plasma creatine concentrations were 34.7+/-15.25 micromol/l in adults, 58.96+/-22.30 micromol/l in children and 5.37, 8.15, 403.5 micromol/l in two untreated children and one treated child with GAMT deficiency, respectively. GAA can also be reliably measured from filter cards, which is sufficient to make the correct diagnosis while creatine is consistently falsely elevated probably secondary to liberation of red cell creatine. In nine healthy newborn infants, GAA concentrations from filter cards were 4.83+/-1.43 and 5.04+/-1.84 micromol/l in 16 healthy adults. We conclude that isotope dilution ES-MS/MS is ideal for rapid high-throughput diagnosis of GAMT deficiency both from plasma and filter paper cards. Using this technique neonatal screening is feasible for this treatable inborn error of creatine metabolism. PMID- 11412831 TI - Simultaneous amplification of DNA and RNA virus using multiplex PCR system. AB - A large number of disease-causing bacteria and viruses are being sequenced and PCR is increasingly used for the diagnosis of the diseases. We have designed a multiplex PCR system for hepatitis B virus (HBV), a DNA virus, and hepatitis E virus (HEV), an RNA virus. A modified technique has been standardized for simultaneous extraction of DNA and RNA, followed by a one-step RT-PCR/PCR. PMID- 11412833 TI - Potent and selective inhibition of the human Na+/H+ exchanger isoform NHE1 by a novel aminoguanidine derivative T-162559. AB - We isolated Na+/H+ exchanger (NHE)-deficient Chinese hamster ovary (CHO-K1) cells stably expressing human NHE isoforms (hNHE1, hNHE2 and hNHE3) and established an assay system for measuring their Na+/H+ exchange activity by monitoring intracellular pH alterations. Using this assay system, we demonstrated that the acylguanidine derivatives, cariporide and eniporide, cause selective inhibition of hNHE1 (IC50 value of 30 nM for cariporide, IC50 value of 4.5 nM for eniporide). Furthermore, we found that a novel synthetic aminoguanidine derivative, T-162559 ((5E,7S)-[7-(5-fluoro-2-methylphenyl)-4-methyl-7,8-dihydro 5(6H)-quinolinylideneamino] guanidine dimethanesulfonate), causes a selective inhibition of hNHE1 with more potent activity than cariporide and eniporide (IC50 value of 0.96 nM). This compound did not affect Na+/HCO3- cotransport and Na+/Ca2+ exchange. PMID- 11412834 TI - Intracellular angiotensin II elicits Ca2+ increases in A7r5 vascular smooth muscle cells. AB - Recent studies show that angiotensin II can act within the cell, possibly via intracellular receptors pharmacologically different from typical plasma membrane angiotensin II receptors. The signal transduction of intracellular angiotensin II is unclear. Therefore, we investigated the effects of intracellular angiotensin II in cells devoid of physiological responses to extracellular angiotensin II (A7r5 vascular smooth muscle cells). Intracellular delivery of angiotensin II was obtained by using liposomes or cell permeabilisation. Intracellular angiotensin II stimulated Ca2+ influx, as measured by 45Ca2+ uptake and single-cell fluorimetry. This effect was insensitive to extracellular or intracellular addition of losartan (angiotensin AT(1) receptor antagonist) or PD123319 ((s)-1 (4-[dimethylamino]-3-methylphenyl)methyl-5-(diphenylacetyl)-4,5,6,7-tetrahydro-1H imidazo[4,5-c]pyridine-6-carboxylate) (angiotensin AT2 receptor antagonist). Intracellular angiotensin II stimulated inositol-1,4,5-trisphosphate (Ins(1,4,5,)P3) production and increased the size of the Ins(1,4,5,)P3 releasable 45Ca2+ pool in permeabilised cells, independent of losartan and PD123319. Small G proteins did not participate in this process, as assessed by using GDPbetaS. Intracellular delivery of angiotensin I was unable to elicit any of the effects elicited by intracellular angiotensin II. We conclude from our intracellular angiotensin application experiments that angiotensin II modulates Ca2+ homeostasis even in the absence of extracellular actions. Pharmacological properties suggest the involvement of putative angiotensin non-AT1-/non-AT2 receptors. PMID- 11412835 TI - Regulation of p42/p44 mitogen-activated protein kinase by the human adenosine A3 receptor in transfected CHO cells. AB - In this study we have investigated whether the human adenosine A3 receptor activates p42/p44 mitogen-activated protein kinase (MAPK) in transfected Chinese hamster ovary (CHO) cells (designated CHO-A3). The high affinity adenosine A3 receptor agonist IB-MECA (1-deoxy-1-[6-[[(3-iodophenyl)methyl]amino]-9H-purin-9 yl]-N-methyl-beta-D-ribofuranuronamide) stimulated time (peak activation occurring after 5 min) and concentration-dependent (pEC50=9.0+/-0.2) increases in p42/p44 MAPK in CHO-A3 cells. Adenosine A3 receptor-mediated increases in p42/p44 MAPK were sensitive to pertussis toxin and the MAPK kinase 1 inhibitor PD 98059 (2'-amino-3'-methoxyflavone). The broad range protein tyrosine kinase inhibitor genistein and the phosphatidylinositol 3-kinase inhibitors wortmannin and LY 294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one) also blocked adenosine A3 receptor stimulation of p42/p44 MAPK. In contrast, inhibition of protein kinase C had no significant effect on adenosine A3 receptor-induced p42/p44 MAPK activation. IB-MECA (pEC50=10.1+/-0.2) also increased the expression of luciferase in CHO-A3 cells transiently transfected with a luciferase reporter gene containing the c-fos promoter. Furthermore, IB-MECA-induced increases in luciferase gene expression were sensitive to pertussis toxin, PD 98059, genistein, wortmannin and LY 294002. In conclusion, we have shown that the human adenosine A3 receptor stimulates p42/p44 MAPK and c-fos-mediated luciferase gene expression in transfected CHO cells. PMID- 11412836 TI - BIA 3-202, a novel catechol-O-methyltransferase inhibitor, enhances the availability of L-DOPA to the brain and reduces its O-methylation. AB - 1-[3,4-Dihydroxy-5-nitrophenyl]-2-phenyl-ethanone (BIA 3-202) is a new long acting catechol-O-methyltransferase (COMT) inhibitor with limited access to the brain. The present study evaluated the interference of BIA 3-202 upon levels of L 3,4-dihydroxyphenylalanine (L-DOPA) and metabolites in plasma (3-O-methyl-L-DOPA) and brain [3-O-methyl-L-DOPA, dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA)] in rats orally treated with L-DOPA (20 mg/kg) plus benserazide (30 mg/kg). At different time points (1, 3 and 6 h) after the administration of BIA 3-202 (0, 3, 10 and 30 mg/kg) or L-DOPA plus benserazide, rats were sacrificed and the right striatum was quickly dissected out and stored for the assay of L-DOPA, 3-O-methyl-L-DOPA, dopamine and amine metabolites. Levels of L-DOPA, 3-O-methyl-L-DOPA, dopamine, DOPAC and HVA in the striatum in L DOPA plus benserazide-treated rats were higher than in vehicle-treated rats. However, this increase in striatal L-DOPA, dopamine, DOPAC and HVA was, in a dose and time-dependent manner, even higher (P<0.05) in rats given BIA 3-202 (3, 10 and 30 mg/kg). This effect was accompanied by a marked decrease in 3-O-methyl-L DOPA levels in the striatum of L-DOPA plus benserazide-treated rats. Increases in levels of L-DOPA and decreases in 3-O-methyl-L-DOPA levels in plasma also accompanied the administration of BIA 3-202. BIA 3-202 did not significantly affect levels of DOPAC and HVA in the striatum in vehicle-treated rats. It is concluded that administration of BIA 3-202 enhances the availability of L-DOPA to the brain by reducing its O-methylation in the periphery, which may prove beneficial in parkinsonian patients treated with L-DOPA plus an aromatic amino acid decarboxylase inhibitor. PMID- 11412837 TI - Anxiolytic effects of aniracetam in three different mouse models of anxiety and the underlying mechanism. AB - The anxiolytic effects of aniracetam have not been proven in animals despite its clinical usefulness for post-stroke anxiety. This study, therefore, aimed to characterize the anxiolytic effects of aniracetam in different anxiety models using mice and to examine the mode of action. In a social interaction test in which all classes (serotonergic, cholinergic and dopaminergic) of compounds were effective, aniracetam (10-100 mg/kg) increased total social interaction scores (time and frequency), and the increase in the total social interaction time mainly reflected an increase in trunk sniffing and following. The anxiolytic effects were completely blocked by haloperidol and nearly completely by mecamylamine or ketanserin, suggesting an involvement of nicotinic acetylcholine, 5-HT2A and dopamine D2 receptors in the anxiolytic mechanism. Aniracetam also showed anti-anxiety effects in two other anxiety models (elevated plus-maze and conditioned fear stress tests), whereas diazepam as a positive control was anxiolytic only in the elevated plus-maze and social interaction tests. The anxiolytic effects of aniracetam in each model were mimicked by different metabolites (i.e., p-anisic acid in the elevated plus-maze test) or specific combinations of metabolites. These results indicate that aniracetam possesses a wide range of anxiolytic properties, which may be mediated by an interaction between cholinergic, dopaminergic and serotonergic systems. Thus, our findings suggest the potential usefulness of aniracetam against various types of anxiety related disorders and social failure/impairments. PMID- 11412838 TI - Effects of 5-HT1A and 5-HT2 receptor agonists on the behavioral and neurochemical consequences of repeated nicotine treatment. AB - This study investigated the effects of repeated daily (15 days) treatment with nicotine, alone or in combination with the 5-HT1A/7 receptor agonist (+/-)-8 hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) or the 5-HT2 receptor agonist (+/-)-2,5-dimethoxy-4-iodoamphetamine (DOI) on locomotor sensitization, mesolimbic dopamine neurochemistry and on behavioral inhibition in the rat. Acute nicotine elevated the extracellular dopamine levels in the nucleus accumbens and stimulated locomotor activity, effects that were sensitized after repeated nicotine treatment. Repeated nicotine administration also produced nicotine induced behavioral disinhibition in the elevated plus-maze. Treatment with DOI counteracted the expression of the nicotine-induced locomotor and neurochemical sensitization, but had no effect on nicotine-induced behavioral disinhibition. Treatment with 8-OH-DPAT decreased the expression of nicotine-induced behavioral disinhibition, but had no effect on locomotor or neurochemical sensitization. Taken together, these findings suggest that the 5-HT1A and the 5-HT2 receptor subtypes are differentially involved in the effects of repeated nicotine on locomotor sensitization, behavioral inhibition and mesolimbic dopamine neurochemistry. PMID- 11412839 TI - The role of cGMP hydrolysing phosphodiesterases 1 and 5 in cerebral artery dilatation. AB - The aim was to investigate the presence and activity of cGMP hydrolysing phosphodiesterases in guinea pig basilar arteries and the effect of selective and non-selective phosphodiesterase inhibitors on cerebral artery dilatation involving the nitric oxide (NO)-guanosine cyclic 3'5-monophosphate (cGMP) pathway. Immunoreactivity to phosphodiesterases 1A, 1B and 5, but not phosphodiesterase 1C was found in fractions of homogenised cerebral arteries eluted by high-pressure liquid chromatography (HPLC). Both the phosphodiesterase 1 inhibitor 8-methoxymethyl-1-methyl-3-(2methylpropyl)-xanthine (8-MM-IBMX) and the phosphodiesterase 5 inhibitors zaprinast and dipyridamole induced dilatation of cerebral arteries. The dilatory response to 8-MM-IBMX was reduced by 1H [1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) (10 microM) and endothelial removal and restored by sodium nitroprusside (0.1 microM) pretreatment, indicating a close relation to the nitric oxide-cGMP pathway. The responses to zaprinast and dipyridamole, however, were not only moderately affected, but also restored by sodium nitroprusside (0.1 microM) pretreatment. At high concentrations, the dilatory effects of zaprinast and dipyridamole were partly caused by cGMP-independent mechanisms. Targeting the phosphodiesterases present in cerebral arteries, with selective inhibitors or activators of phosphodiesterase, may be a possible new way of treating cerebrovascular disease. PMID- 11412840 TI - Modifications by sumatriptan and acetylcholine of nitric oxide-mediated neurogenic dilatation in dog cerebral arteries. AB - Canine cerebral arterial strips denuded of endothelium responded to nicotine and transmural electrical stimulation with relaxations, which were abolished by NG nitro-L-arginine and methylene blue. Magnitudes of relaxation did not differ in the arteries contracted with prostaglandin F2alpha and sumatriptan, an effective therapeutic of migraine. Sumatriptan concentration-dependently contracted the arteries responding to 2 Hz stimulation with persistent relaxations, and the concentration of this 5-HT1B/1D/1F receptor agonist to overcome the relaxation averaged 1.06 x 10(-7) M. Acetylcholine inhibited the response to nerve stimulation due possibly to its action on prejunctional nitroxidergic nerves; the inhibition did not differ in the arteries contracted with prostaglandin F2alpha and K+. It appears that sumatriptan does not interfere with the release of nitric oxide from nerves but counteracts the neurogenic relaxation by functional antagonistic action on smooth muscle. Prejunctional inhibition by muscarinic receptor activation is unlikely associated with opening of neuronal K+ channels. PMID- 11412841 TI - E-BioSci. PMID- 11412842 TI - The 14th Datta Lecture. TFIIH: from transcription to clinic. AB - Once a large proportion of the genes responsible for genetic disorders are identified in the post-genome era, the fundamental challenge is to establish a genotype/phenotype relationship. Our aim is to explain how mutations in a given gene affect its enzymatic function and, in consequence, disturb the life of the cell. Genome integrity is continuously threatened by the occurrence of DNA damage arising from cellular exposure to irradiation and genotoxic chemicals. This mutagenic or potentially lethal DNA damage induces various cellular responses including cell cycle arrest, transcription alteration and processing by DNA repair mechanisms, such as the nucleotide excision repair (NER) pathway. Disruption of NER in response to genotoxic injuries results in autosomal recessive hereditary diseases such as Xeroderma pigmentosum (XP), Cockayne syndrome (CS) and trichothiodystrophy (TTD). One of the most immediate consequences of the induction of strand-distorting lesions is the arrest of transcription in which TFIIH plays a role in addition to its role in DNA repair. The observations made by clinicians close to XP, TTD and CS patients, suggested that transcription defects responsible for brittle hair and nails for TTD, or developmental abnormalities for CS, resulted from TFIIH mutations. Here a story will be related which could be called 'a multi-faceted factor named TFIIH'. As biochemists, we have characterized each component of TFIIH, three of which are XPB and XPD helicases and cdk7, a cyclin-dependent kinase. With the help of structural biologists, we have characterized most of the specific three dimensional structures of TFIIH subunits and obtained its electron microscopy image. Together these approaches help us to propose a number of structure function relationships for TFIIH. Through transfection and microinjection assays, cell biology allows us to determine the role of TFIIH in transcription and NER. We are thus in a position to explain, at least in part, transcription initiation mechanisms and their coupling to DNA repair. We now know how the XPB helicase opens the promoter region for RNA synthesis and that one of the roles of XPD helicase is to anchor the cdk7 kinase to the core-TFIIH. In XP and CS associated patients, we have demonstrated that some XPD mutations prevent an optimal phosphorylation of nuclear receptors by cdk7 with, as a consequence, a drop in the expression of genes sensitive to hormone action. We have thus shown that hormonal responses operate through TFIIH. Careful analysis of each TFIIH subunit also shows how the p44 Ring finger participates in certain promoter escape reactions. We are also able to localize the action of TFIIH in the sequence of events that lead to the elimination of DNA lesions. Thanks to the combination of these different approaches we are obtaining a much clearer picture of the TFIIH complex and its integration into the life of the cell. PMID- 11412843 TI - PABMB Lecture. Protein dynamics, folding and misfolding: from basic physical chemistry to human conformational diseases. AB - Proteins exhibit a variety of motions ranging from amino acid side-chain rotations to the motions of large domains. Recognition of their conformational flexibility has led to the view that protein molecules undergo fast dynamic interconversion between different conformational substates. This proposal has received support from a wide variety of experimental techniques and from computer simulations of protein dynamics. More recently, studies of the subunit dissociation of oligomeric proteins induced by hydrostatic pressure have shown that the characteristic times for subunit exchange between oligomers and for interconversion between different conformations may be rather slow (hours or days). In such cases, proteins cannot be treated as an ensemble of rapidly interconverting conformational substates, but rather as a persistently heterogeneous population of different long-lived conformers. This is reminiscent of the deterministic behavior exhibited by macroscopic bodies, and may have important implications for our understanding of protein folding and biological functions. Here, we propose that the deterministic behavior of proteins may be closely related to the genesis of conformational diseases, a class of pathological conditions that includes transmissible spongiform encephalopathies, Alzheimer's disease and other amyloidosis. PMID- 11412844 TI - Mutualists and parasites: how to paint yourself into a (metabolic) corner. AB - Eukaryotes have developed an elaborate series of interactions with bacteria that enter their bodies and/or cells. Genome evolution of symbiotic and parasitic bacteria multiplying inside eukaryotic cells results in both convergent and divergent changes. The genome sequences of the symbiotic bacteria of aphids, Buchnera aphidicola, and the parasitic bacteria of body louse and humans, Rickettsia prowazekii, provide insights into these processes. Convergent genome characteristics include reduction in genome sizes and lowered G+C content values. Divergent evolution was recorded for amino acid and cell wall biosynthetic genes. The presence of pseudogenes in both genomes provides examples of recent gene inactivation events and offers clues to the process of genome deterioration and host-cell adaptation. PMID- 11412845 TI - Transcriptomes, transcription activators and microarrays. AB - Gene-specific transcription activators are among the main factors which specifically shape the transcriptome profiles. It is tempting to take advantage of their properties to decipher the genome expression circuitry. The advent of microarray technology has offered fantastic opportunities to quickly analyze the expression profiles dictated by specific transcription factors. This review will first focus on the strategies which have been devised to control the activity of transcription factors and in the second part on the microarray experiments which addressed the role of these transcription factors in the genome-wide expression profile. This last part will mainly consider the case of the yeast Saccharomyces cerevisiae genome. All the collected data are available through the on-line database yTAFNET (http://transcriptome.ens.fr/ytafnet/). yTAFNET is designed to help the characterization of connections between the different yeast regulatory networks. PMID- 11412846 TI - Molecular mechanism of translocation through nuclear pore complexes during nuclear protein import. AB - The trafficking of macromolecules between cytoplasm and nucleus through nuclear pore complexes is mediated by specific carrier molecules such as members of the importin-beta family. Nuclear pore proteins (nucleoporins) frequently contain sequence repeats based on FG cores and carriers appear to move their cargo through the pores by hopping between successive FG cores. A major question is why some macromolecules are transported while others are not. This selectivity may be generated by the ability to bind FG repeats, a local concentration of carrier cargo complexes near the entrance to the pore channel, and steric hindrance produced by high concentrations of nucleoporins in the channel. PMID- 11412847 TI - Nuclear export of mRNA. AB - Export of mRNA through nuclear pore complexes (NPC) is preceded by multiple and well coordinated processing steps, resulting in the formation of an export competent ribonucleoprotein complex (mRNP). Numerous factors involved in the translocation of the mRNP through the NPC and its release into the cytoplasm have been isolated mainly through genetic approaches in yeast, and putative functional homologues have been identified in metazoan systems. Understanding the mechanism of mRNA export relies, in part, on the functional characterization of these factors and the establishment of a complete network of molecular interactions. Here we summarize recent progress in the characterization of yeast and mammalian components implicated in the export of an mRNA from the nucleus to the cytoplasm. PMID- 11412848 TI - The rules and roles of nucleocytoplasmic shuttling proteins. AB - The spatial separation of mRNA synthesis from translation, while providing eukaryotes with the possibility to achieve higher complexity through a more elaborate regulation of gene expression, has set the need for transport mechanisms through the nuclear envelope. In a simplistic view of nucleocytoplasmic transport, nuclear proteins are imported into the nucleus while RNAs are exported to the cytoplasm. The reality is, however, that transport of either proteins or RNAs across the nuclear envelope can be bi-directional. During the past years, an increasing number of proteins have been identified that shuttle continuously back and forth between the nucleus and the cytoplasm. The emerging picture is that shuttling proteins are key factors in conveying information on nuclear and cytoplasmic activities within the cell. PMID- 11412849 TI - A role for Cajal bodies in assembly of the nuclear transcription machinery. AB - Cajal bodies (CBs) are small nuclear organelles that contain the three eukaryotic RNA polymerases and a variety of factors involved in transcription and processing of all types of RNA. A number of these factors, as well as subunits of polymerase (pol) II itself, are rapidly and specifically targeted to CBs when injected into the cell. It is suggested that pol I, pol II, and pol III transcription and processing complexes are preassembled in the CBs before transport to the sites of transcription on the chromosomes and in the nucleoli. PMID- 11412850 TI - Rho proteins, PI 3-kinases, and monocyte/macrophage motility. AB - Rho proteins and phosphatidylinositide 3-kinases (PI 3-kinases) have been widely implicated in regulating cell motility both in cultured cells and in animal models. Monocytes are recruited from the bloodstream in response to inflammatory signals, and migrate across the endothelial barrier into the tissues, where they differentiate into macrophages and phagocytose bacteria and cells. Studies of monocytes and macrophages have revealed that different Rho family members and PI 3-kinases are not functionally redundant but play unique and distinct roles in motile responses. PMID- 11412851 TI - Osmotic stress activates distinct lipid and MAPK signalling pathways in plants. AB - Plants are continuously exposed to all kinds of water stress such as drought and salinity. In order to survive and adapt, they have developed survival strategies that have been well studied, but little is known about the early mechanisms by which the osmotic stress is perceived and transduced into these responses. During the last few years, however, a variety of reports suggest that specific lipid and MAPK pathways are involved. This review briefly summarises them and presents a model showing that osmotic stress is transmitted by multiple signalling pathways. PMID- 11412852 TI - Coordination between transcription and pre-mRNA processing. AB - A large body of work has proved that transcription by RNA polymerase II and pre mRNA processing are coordinated events within the cell nucleus. Capping, splicing and polyadenylation occur while transcription proceeds, suggesting that RNA polymerase II plays a role in the regulation of these events. The presence and degree of phosphorylation of the carboxy-terminal domain of RNA polymerase II large subunit is important for functioning of the capping enzymes, the assembly of spliceosomes and the binding of the cleavage/polyadenylation complex. Nuclear architecture and gene promoter structure have also been shown to play key roles in coupling between transcription and splicing. PMID- 11412853 TI - Oxidative stress, aging and longevity in Drosophila melanogaster. AB - Free radicals produced during normal metabolism cause damage to macromolecules. The free radical theory of aging proposes that the organism is unable to repair all of them and that, with time, unrepaired damages accumulate and put the organism at risk: in other words, free radicals provoke aging and death. This article reviews both the results of adding antioxidants to food on longevity in Drosophila melanogaster, as well as the studies on antioxidant enzymes (inactivation in vivo, null mutants, overexpression). It is concluded that antioxidant enzymes are probably poorly connected to the normal aging process, but they allow the organism to cope with stressful conditions. PMID- 11412854 TI - Role of AP2/EREBP transcription factors in gene regulation during abiotic stress. AB - Crop plants are exposed to many types of abiotic stress during their life cycle. Water deficit derived from drought, low temperature or high salt concentration in the soil, is one of the most common environmental stresses that affects growth and development of plants through alterations in metabolism and gene expression. Adaptation to these conditions may involve passive tolerance or active homeostatic mechanisms for maintaining water balance. Active responses occur at different levels in the plant and may represent a concomitant protection against other types of stress such as pathogen attack. Many morphological and physiological adaptations to water stress are under the control of the plant hormone abscisic acid and involve specific activation of target genes that in one way or another protect cells against water deficit or participate in the regulation of the drought response. Here, we discuss recent advances in our understanding of drought adaptation mediated by specific changes in gene expression and the role of AP2/EREBP nuclear factors in these processes. PMID- 11412855 TI - Endocytosis and signaling cascades: a close encounter. AB - Internalization of receptors and other cell surface components is well known to occur via clathrin-mediated endocytosis, although other less well characterized pathways are also involved. Internalized receptors are then delivered to early endosomes, where they are sorted to be recycled back to the plasma membrane for reutilization or transported to late endosomes/lysosomes for degradation. Endocytosis has long been considered as a constitutive, housekeeping function of animal cells that occurs independently of the cellular environment in contrast to regulated secretion. Here, we will discuss recent studies that are uncovering the existence of cross-talk between signaling molecules and components of the transport machinery, indicating that endocytosis can be modulated by signaling pathways. PMID- 11412856 TI - Prenylation of Rab GTPases: molecular mechanisms and involvement in genetic disease. AB - Small GTPases of the Rab family regulate membrane transport pathways. More than 50 mammalian Rab proteins are known, many with transport step-specific localisation. Rabs must associate with cellular membranes for activity and membrane attachment is mediated by prenyl (geranylgeranyl) post-translational modification. Mutations in genes encoding proteins essential for the geranylgeranylation reaction, Rab escort protein and Rab geranylgeranyl transferase, underlie genetic diseases. Choroideremia patients have loss of function mutations in REP1 and the murine Hermansky-Pudlak syndrome model gunmetal possesses a splice-site mutation in the alpha-subunit of RGGT. Here we discuss recent insights into Rab prenylation and advances towards our understanding of both diseases. PMID- 11412857 TI - Transthyretin amyloidosis: a tale of weak interactions. AB - Over 70 transthyretin (TTR) mutations have been associated with hereditary amyloidoses, which are all autosomal dominant disorders with adult age of onset. TTR is the main constituent of amyloid that deposits preferentially in peripheral nerve giving rise to familial amyloid polyneuropathy (FAP), or in the heart leading to familial amyloid cardiomyopathy. Since the beginning of this decade the central question of these types of amyloidoses has been why TTR is an amyloidogenic protein with clinically heterogeneous pathogenic consequences. As a result of amino acid substitutions, conformational changes occur in the molecule, leading to weaker subunit interactions of the tetrameric structure as revealed by X-ray studies of some amyloidogenic mutants. Modified soluble tetramers exposing cryptic epitopes seem to circulate in FAP patients as evidenced by antibody probes recognizing specifically TTR amyloid fibrils, but what triggers dissociation into monomeric and oligomeric intermediates of amyloid fibrils is largely unknown. Avoiding tetramer dissociation and disrupting amyloid fibrils are possible avenues of therapeutic intervention based on current molecular knowledge of TTR amyloidogenesis and fibril structure. PMID- 11412858 TI - Protein misfolding and disease; protein refolding and therapy. AB - Diverse human disorders, including several neurodegenerative diseases and systemic amyloidosis, are thought to arise from the misfolding and aggregation of an underlying protein. Recent findings strongly support this hypothesis and have increased our understanding of the molecular mechanism of protein conformational disorders. Many questions are still pending, but the data overall suggest that correction of protein misfolding constitutes a viable therapeutic strategy for conformational diseases. PMID- 11412859 TI - The catalytic mechanism of protein tyrosine phosphatases revisited. AB - Experimental and theoretical studies of the catalytic mechanism in protein tyrosine phosphatases and dual specific phosphatases are reviewed. The structural properties of these enzymes contributing to the efficient rate enhancement of phosphate monoester hydrolysis have been established during the last decade. There are, however, uncertainties in the interpretation of available experimental data that make the commonly assumed reaction mechanism somewhat doubtful. Theoretical calculations as well as analysis of crystal structures point towards an alternative interpretation of the ionisation state in the reactive complex. PMID- 11412860 TI - A genomic perspective on human proteases. AB - Over 400 human proteases documented in secondary databases can already be delineated in genomic sequence. A Genome Ontology annotation of 30585 sequences in the provisional human proteome set recognises 498 proteases, i.e. 1.6%. Homology searches against finished sequence and comparisons between mouse and zebrafish are likely to increase this total. However, the data already indicate that the mechanistic class, sequence family and domain distribution of the genomic complement of proteases is unlikely to shift significantly from that already observed in the transcript data. Genomically derived novel sequences will require bioinformatic analysis and biochemical verification. The increasing availability of annotated genomic data will enable studies of splice variants, transcriptional control, polymorphisms, pseudogenes, inactive homologues and evolution. Comparative work on complete human protease families should produce a more integrated picture of their biochemistry and physiology. Genomic data will also lead to the identification of new protease involvement in disease processes and their evaluation as drug targets. PMID- 11412861 TI - Antibiotic activities of peptides, hydrogen peroxide and peroxynitrite in plant defence. AB - Genes encoding plant antibiotic peptides show expression patterns that are consistent with a defence role. Transgenic over-expression of defence peptide genes is potentially useful to engineer resistance of plants to relevant pathogens. Pathogen mutants that are sensitive to plant peptides in vitro have been obtained and a decrease of their virulence in planta has been observed, which is consistent with their hypothetical defence role. A similar approach has been followed to elucidate the potential direct anti-microbial role of hydrogen peroxide. Additionally, a scavenger of peroxynitrite has been used to investigate its involvement in plant defence. PMID- 11412862 TI - What can yeast tell us about N-linked glycosylation in the Golgi apparatus? AB - The N-glycans found on eukaryotic glycoproteins occur in a vast range of different structures. A universal N-glycan core is attached to proteins during synthesis in the endoplasmic reticulum, and then diversity is generated as the proteins pass through the Golgi apparatus. Many of the Golgi-localised glycosyltransferases have now been identified in both yeast and mammalian cells, but it is still unclear how these enzymes are integrated into the Golgi and the rest of the cell so as to ensure efficient and specific processing of passing substrates. This review discusses the potential of the yeast system to address these issues. PMID- 11412864 TI - Helicobacter pylori infection as an independent risk factor for cerebral ischemia of atherothrombotic origin. AB - Chronic infection may increase the risk for ischemic stroke. Presently, it is insufficiently established whether Helicobacter pylori infection represents a risk factor for ischemic stroke. We analyzed IgG antibodies against H. pylori in 109 patients with acute cerebral ischemia and 82 age- and sex-matched control patients with non-vascular and non-inflammatory neurological diseases. Antibody titers were significantly higher in patients than in control subjects (p=0.007). H. pylori seropositivity tended to be more common in patients (odds ratio (OR) 1.55, 95% confidence interval (ci) 0.87-2.76), but this trend was further attenuated in multivariate analysis (OR 1.42; 95% 0.75-2.67) with hypertension, diabetes mellitus, current or previous smoking, previous cerebral ischemia and low socioeconomic status. H. pylori seropositivity increased the odds for cerebral ischemia of atherothrombotic origin in univariate (OR 3.63; 95% ci 1.37 9.65) and multivariate analysis (OR 3.53; 95% ci 1.09-11.4). H. pylori seropositivity may be an independent risk factor for stroke of atherothrombotic origin. PMID- 11412865 TI - REM sleep behavior disorder in sleep-disordered patients with versus without Parkinson's disease: is there a need for polysomnography? AB - We reviewed the polysomnograms (PSGs) of 292 consecutive patients with sleep disorders (Parkinson's disease (PD), n=19, other sleep disorders, n=273) to investigate the sensitivity and specificity of the clinical diagnosis of rapid eye movement behavior disorder (RBD) compared with polysomnographic diagnosis. Patients with dementia, multiple system atrophy, or any other neurodegenerative disease were excluded. RBD was diagnosed clinically if the minimal criteria, according to the guidelines given in the International Sleep Disorders Classification, were fulfilled. The following PSG criteria were required for diagnosis of RBD: REM sleep without muscle atonia seen in PSG associated with motor behavior visible in the PSG-synchronized videotape. Nine of nineteen PD patients (47%) had RBD. RBD occurred in only four patients without PD (1.8%). The sensitivity of specialized interviews for identifying RBD clinically was good in non-PD patients (sensitivity: 100%, specificity: 99.6%). However, the sensitivity was poor (33%) with a specificity of 90%, in patients with PD. We conclude that the diagnosis of RBD in patients with PD requires PSG, whereas interviews are sufficient for diagnosing RBD in non-PD patients. PMID- 11412866 TI - A combined study of tumor-related brain lesions using MEG and proton MR spectroscopic imaging. AB - The purpose of this study is to localize, in cases of brain tumors, pathological magnetic brain activities and to analyze metabolic alterations in functionally abnormal lesions using magnetoencephalography (MEG) and proton magnetic resonance spectroscopic imaging (1H MRSI). The study focused on 10 healthy volunteers and seven patients with common brain tumors, namely astrocytic tumor and meningioma. In spontaneous MEG, the pathological brain activities (slow, fast waves and spikes) were localized using a single equivalent dipole model. After the results of MEG and 1H MRSI were superimposed onto the corresponding MR images, the signal intensities of spectroscopically visible metabolites were analyzed in the regions where the dipoles of the pathological activities were concentrated. Increased slow wave activity was observed in four cases and fast wave or spike activity was significantly increased in one case. These pathological activities were localized in surrounding regions of the bulk of tumors, where mild reduction of N-acetyl aspartate (NAA) and slight accumulation of lactate (Lac) consistently existed. Preserved cortical areas, which are indicated by residual NAA, might be able to generate pathological magnetic activities under lactic acidosis. Such areas could be understood as a border zone between normal and seriously damaged brain tissue by tumors or associated brain edema. This combined technique with the different modalities gives insight into functional as well as metabolic aspects of pathological brain conditions. PMID- 11412867 TI - Granulocyte integrins before and after activation in acute ischaemic stroke. AB - We examined in 19 subjects with acute ischaemic stroke (AIS) the PMN integrin pattern (CD11a, CD11b, CD11c, CD18), using indirect immunofluorescence and adopting a flow cytometer, at baseline and during activation, prolonged for 5 and 15 min, with 4-phorbol 12-myristate 13-acetate (PMA). At baseline, an increase in the expression of CD11c and CD18 and a decrease in the CD11b were evident in AIS subjects compared to normals. After activation, we found in normals a constant and significant increase of all PMN adhesive molecules, while in AIS subjects, we found an increase in CD11b and CD18, a decrease in CD11a and no variation in CD11c. While the basal upregulation of CD11c and CD18 may depend on the PMN spontaneous activation or on the increase of cytokines, the decrease of CD11b may be due to its self-consumption. After activation, the decrease in CD11a noted in AIS may be related to its cleavage or to an altered integrin phosphorylation/dephosphorylation balance. PMID- 11412868 TI - Significance of serotonin transporter gene polymorphism in migraine. AB - OBJECTIVE: To elucidate significance of the serotonin transporter gene (STG) polymorphism in migraine, and to address the polymorphic patterns of STG, both in the migraineurs and healthy people in this country. STUDY DESIGN: A PCR study of STG in 52 migraineurs and 80 healthy controls. METHODS: Using the PCR technique, STG polymorphism was studied in the DNA obtained from leukocytes of the patients and healthy controls. Polymorphism of the two regions (VNTR and 5-HTTLPR) of STG was assessed. RESULTS: VNTR STin 2.10 and STin 2.12 alleles were detected in migraineurs and healthy controls. Both homozygous and heterozygous STin 2.10 allele predominated in the migraine group (p=0.01), while STin 2.12 allele was more frequent in the healthy controls (p=0.02). There was no relationship between the migraine type, family history of migraine and STG polymorphism. CONCLUSION: STin 2.10 and STin 2.12 alleles of VNTR are frequent in this country. While the presence of STin 2.10 allele increases the risk of migraine, 5-HTTLPR polymorphism is not associated with this risk. PMID- 11412869 TI - Apoptosis in neurones exposed to cerebrospinal fluid from patients with multiple sclerosis or acute polyradiculoneuropathy. AB - Primary cultures of murine cerebellar granule neurones were exposed to cerebrospinal fluid from patients with subtypes of multiple sclerosis or acute polyradiculoneuropathy (Guillain-Barre syndrome) for 2 days. Cells were then stained with Hoechst 33342 or terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) to detect apoptotic bodies. The results were compared with control cultures exposed to cerebrospinal fluid from patients with no known neurological disease or deficit. There was no significant difference in the level of apoptosis induced between these controls and cultures not exposed to cerebrospinal fluid at all. Cultures exposed to cerebrospinal fluid samples from patients with relapsing-remitting multiple sclerosis did not have higher levels of apoptosis than cells exposed to controls, regardless of whether the sample was taken during relapse or remission. However, a significant increase in apoptosis was observed in cultures exposed to cerebrospinal fluid from patients with primary progressive multiple sclerosis, and apoptosis correlated with disease severity. This supports the existence of biochemical differences between subgroups of multiple sclerosis. A significant increase in apoptosis was also induced by cerebrospinal fluid samples from patients with acute polyradiculoneuropathy, suggesting the presence of neurotoxic factor(s) here also. The relevance to disease pathology is unclear. PMID- 11412870 TI - MRI-FLAIR images of the head show corticospinal tract alterations in ALS patients more frequently than T2-, T1- and proton-density-weighted images. AB - In some patients with amyotrophic lateral sclerosis (ALS), T2-weighted and proton density-weighted magnetic resonance imaging (MRI) shows hyperintense or hypointense signals at the corticospinal tract. Fluid-attenuated inversion recovery (FLAIR) sequences increase the sensitivity of MRI to detect cortical and subcortical tissue changes. In 31 ALS patients and 33 controls, we studied the frequency and the extent of signal abnormalities in FLAIR images compared to T2-, T1- and proton-density-weighted images. Hyperintense signals at the corticospinal tract were significantly more frequent in FLAIR images than in all other tested sequences. In FLAIR images of ALS patients only, distinct hyperintense signals at the subcortical precentral gyrus (five patients), the centrum semiovale (eight patients), the crus cerebri (nine patients) and the pons (four patients) as well as mild hyperintense signals in the medulla oblongata (three patients) were seen. More frequently, but not exclusively in ALS patients, FLAIR images showed mild hyperintense signals at the subcortical precentral gyrus (15 patients vs. 1 control). Quantitative analysis confirmed the significant difference between ALS patients and controls at the subcortical precentral gyrus in FLAIR images. In T1 weighted images, the corticospinal tract at the capsula interna was hypointense in significantly more controls than ALS patients. Also this difference was confirmed in the quantitative analysis. Similar to previous results, MR image alterations did correlate poorly to clinical data of upper motor neuron affliction.MR images of the head, including FLAIR images, provide additional information regarding corticospinal tract involvement in ALS patients. Because of an overlap with physiological findings, they have to be interpreted cautiously, with the exception of hyperintense signals at the subcortical precentral gyrus. PMID- 11412871 TI - Cardiovascular autonomic control in Becker muscular dystrophy. AB - Although autonomic symptoms are not prominent in dystrophinopathies, a reduced vagal activity and an enhanced sympathetic tone have been found in Duchenne muscular dystrophy. Twenty patients with Becker muscular dystrophy (BMD) were investigated by a battery of six cardiovascular autonomic tests (beat-to-beat variability during quiet breathing and deep breathing, heart rate responses to Valsalva maneuver and standing, blood pressure responses to standing and sustained handgrip) and power spectral analysis (PSA) of heart rate variability. Although 11 patients revealed abnormal findings at some cardiovascular tests, none of them had a definite autonomic damage, as indicated by two or more abnormal tests. The mean results of the single tests did not differ from normal controls, except for the beat-to-beat variability during quiet breathing, which was significantly higher in BMD (p<0.05). Such finding was confirmed by a significantly higher total variance (p<0.05), indicating an increased parasympathetic activity. Spectral components were not significantly different from normal controls. PSA values were not influenced by age, functional ability score or presence of heart abnormalities. Our data suggest that autonomic involvement does not represent a major finding in BMD. PMID- 11412872 TI - Transcriptional activation of the non-muscle, full-length dystrophin isoforms in Duchenne muscular dystrophy skeletal muscle. AB - Despite promoter tissue specificity, up-regulation of the brain and Purkinje cell type dystrophin isoforms was described in skeletal muscle of X-linked dilated cardiomyopathy (XLDCM) and BMD affected individuals. An extended population of 11 Duchenne muscular dystrophy (DMD) and 11 Becker muscular dystrophy (BMD) patients was investigated to determine whether ectopic muscle expression of the two full length non-muscular isoforms is a common event in dystrophinopathies and if it has functional significance. Up-regulation of the two non-muscle-specific isoforms was detected in four DMD patients but in none of the BMD affected individuals or non-dystrophic controls. This is the first report of an expression of these two isoforms in DMD skeletal muscle. Ectopic expression is not confined to regenerating or revertant fibers and does not correlate with age at biopsy, clinical phenotype, cardiac involvement, deletion size or location. We consider that muscle ectopic expression of the brain and Purkinje cell-type isoforms has no favorable prognostic significance in DMD and BMD patients. PMID- 11412873 TI - Sensory neuropathy of the trigeminal, glossopharyngeal, and vagal nerves in Sjogren's syndrome. AB - Isolated cranial nerve involvement in primary Sjogren's syndrome (primary SS) has rarely been described. We report the case of a patient with sensory neuropathy of the trigeminal and also the glossopharyngeal and vagal nerves, which has not been identified previously. The electrophysiological findings in our patient with primary SS confirmed trigeminal sensory neuropathy with abnormal blink reflexes and abnormal cutaneous masseter inhibitory reflexes. PMID- 11412874 TI - LFA-1 expression on CD4(+)CD45RO(+) peripheral blood T-lymphocytes in RR MS: effects induced by rIFNbeta-1a. AB - We investigate the in vivo and in vitro effects of short-term treatment with recombinant Interferon beta-1a (rIFNbeta-1a) on CD4(+)CD45RO(+) activated/memory peripheral blood T-lymphocytes (PBTLs) expressing Leukocyte Function Antigen-1 (LFA-1; CD11a/CD18) in relapsing-remitting (RR) Multiple Sclerosis (MS) patients. Blood samples were obtained from 10 RR MS patients before and after 2, 4 and 6 months of rIFNbeta-1a (Avonex) treatment. For each sample, the percentage of CD4(+)CD45RO(+)CD11a(+) (CD11a(dim) and CD11a(bright)) T-cells was evaluated in in vivo PBTLs and in untreated or rIFNbeta-1a (1000 U/ml) or recombinant soluble Intercellular Adhesion Molecule-1 (ICAM-1, the ligand for LFA-1) (400 ng/ml) treated cultured PBTLs by triple fluorescence flow-cytometry (FACS analysis). Soluble ICAM-1 (sICAM-1) serum levels were evaluated by ELISA. In vivo, the percentage of CD4(+)CD45RO(+), CD4(+)CD45RO(+)CD11a(+), CD4(+)CD45RO(+)CD11a(dim) PBTLs increased after 4 and 6 months of rIFNbeta-1a treatment compared to pretreatment and 2 months of treatment (p<0.05). The CD11a expression per se did not change during the time course. Soluble ICAM-1 (sICAM-1) serum levels also increased (p<0.05) after 4 and 6 months of treatment. When T-cells, obtained from the blood of the same patients before and during in vivo treatment, were cultured either untreated or treated with rIFNbeta-1a, they showed an increase in the percentage of CD4(+)CD45RO(+) T-cells expressing CD11a(bright) (p<0.05). The addition of recombinant sICAM-1 to untreated cultures decreased the percentage of CD4(+)CD45RO(+) T-cells expressing CD11a. This last finding seems to support an indirect effect in vivo of rIFNbeta-1a via sICAM-1 on this T-cell subset, since the ICAM-1 soluble form, induced in vivo in serum by rIFNbeta-1a but lacking in in vitro conditions, keeps the percentage of CD11a(+) unchanged within CD4(+)CD45RO(+) T-cells and induces their expression of CD11a(dim), probably preventing T-cells from transmigrating. PMID- 11412875 TI - Expression of vascular endothelial growth factor in tuberculous meningitis. AB - The pathogenesis of tuberculous meningitis is still unclear. Recently, vascular endothelial growth factor (VEGF) was found to be associated with inflammatory diseases and we found the increased serum level of VEGF in pulmonary tuberculosis. We hypothesized that VEGF might be associated with the pathogenesis of tuberculous meningitis and measured serum and cerebrospinal fluid (CSF) levels of VEGF in 28 patients with tuberculous meningitis and 31 non-tuberculous infectious meningitis patients (13 bacterial meningitis patients, eight fungal meningitis patients and 10 patients with viral meningitis) before therapy. We examined the CSF VEGF levels 3 months after in 12 tuberculous meningitis patients. The serum and CSF levels of VEGF were significantly higher in tuberculous meningitis than in other meningitis. The decrease in titer of CSF VEGF paralleled the clinical improvement of tuberculous meningitis. Immunohistochemical staining of autopsied brains demonstrated the presence of VEGF in the inflammatory mononuclear cells of the dense fibroconnective tissue both in the subarachnoid space and surrounding the vasculitis lesion. We found the expression of VEGF in tuberculous meningitis and think that VEGF reflects its activity. PMID- 11412876 TI - Inflammatory polyradiculoneuropathy with spinal cord involvement and lethal [correction of letal] outcome after hepatitis B vaccination. AB - We report on a 36-year-old man who developed an inflammatory polyradiculoneuropathy similar to Guillain-Barre syndrome 9 days after hepatitis B vaccination. Extensive immunotherapy including immunoglobulins, steroids, plasmapheresis, cyclophosphamide and methotrexate did not stop the progressive course of the disease and the patient died 4 months later due to multiorgan failure with septic shock symptoms and adult respiratory distress syndrome (ARDS).The neuropathological investigation showed severe axonal loss with mild demyelination of peripheral nerves and mononuclear cell infiltrates, predominantly T-lymphocytes, in nerve roots and spinal ganglia. In addition, there were unusual, perivascular and parenchymal lymphocytic cell infiltrates in the grey matter, especially the anterior horns of the spinal cord. The temporal relationship to hepatitis B vaccination, the strong increase of HBs-antibodies within 3 weeks after vaccination, and the presumptive immune mediated pathology of this disorder suggest a possible etiologic link with hepatitis B vaccine. PMID- 11412877 TI - Differential cellular expression of tumor necrosis factor-alpha and Type I tumor necrosis factor receptor after transient global forebrain ischemia. AB - We examined the expression of tumor necrosis factor-alpha (TNF-alpha) and the Type I tumor necrosis factor receptor, (TNFR1), in relation to c-fos, a known regulator gene of immediate cellular responses, after an extended period of global ischemia. The number of TNF-alpha mRNA expressing cells peaked in most brain areas after 8 h of reperfusion. Significant increases in TNFR1 mRNA expression were evident in the cortex at 2 and 8 h of reperfusion and after 8 h of reperfusion in the CA3/CA4 region of the hippocampus. Transient neuronal c-fos mRNA expression preceded these responses. TNF-alpha immunoreactivity was seen in neurons>>>oligodendrocytes=perivascular cells=ependymal cells=vessel wall structures. After ischemia/reperfusion, increased TNF-alpha immunoreactivity was evident only in oligodendrocytes. TNFR1 immunoreactivity in sham brains manifested in bundles of cellular fibers of variable length and thickness. In post-ischemic brains, immunoreactivity in these cellular processes representing mainly astroglial extensions was suppressed at 2 h but recovered partially by 8 and 24 h of reperfusion. In contradiction, transient ischemia-induced TNFR1 immunoreactivity was observed in somas of large cortical neurons, in activated microglia/macrophages, perivascular and endothelial cells.Taken together, the increase in neuronal TNF-alpha mRNA appeared not to be followed by substantial translation to protein in the cerebral tissue after an extended period of global ischemia. However, there was increased neuronal TNFR1 immunostaining in conjunction with increased immunostaining for TNF-alpha in oligoglial elements, which suggests signaling to neurons by enhanced oligoglial TNF-alpha. PMID- 11412878 TI - H reflex behavior in Parkinson's disease patients and patients with extrapyramidal and pyramidal signs combined. AB - The H reflex was elicited in 21 normal subjects, 48 patients with Parkinson's disease (PD) and 22 patients with pyramidal and extrapyramidal signs combined (PESC). In most normal subjects (90.5%), in 29.2% of PD patients and 54.5% of patient with PESC the threshold for sensory fibers was lower than for motor fibers, and the H reflex was obtained before the M response for all duration stimuli in both legs. In 9.5% of normal subjects, 39.6% of PD patients with mild and moderate rigidity (according to the motor part of UPDRS) and 31.8% of patients with PESC, the threshold for the H reflex and M response was the same or the M response threshold was lower in at least one of the legs for short stimulus duration (0.1-0.2 ms). In 31.2% of PD patients (most of them with severe rigidity) and 13.7% of patients with PESC, the threshold for M response was lower for all stimulus duration in at least one of the legs, and it was obtained before H reflex. These very significant differences in behavior of the H reflex in PD patients (Fisher exact test, p<0.0001) that almost disappear in patients with PESC, could be possibly explained by changes in agonist-antagonist inhibition. PMID- 11412879 TI - Color vision deficiencies in Gilles de la Tourette syndrome. AB - Color perception was tested using the Farnsworth-Munsell 100-Hue Test in a sample of persons with Gilles de la Tourette syndrome (GTS), and compared to norms from three age cohorts in the early second, fourth and sixth decades. Red-green color errors on the Farnsworth-Munsell did not appear to change appreciably as a function of age or GTS. Blue-yellow error scores did, however, increase with age and were exaggerated in the GTS group. It is concluded that sensory and perceptual disturbances are present in GTS as in other basal cell ganglia disorders. The results are discussed in terms of converging retinal dopaminergic mechanisms also associated with Parkinson's and Huntington's diseases and even with normal aging. Suggestions are offered that daily activities and behavior may be affected by spatial and chromatic deficiencies. PMID- 11412880 TI - Contrast gain mechanism or transient channel? Why the effects of a background pattern alter over time. AB - If a brief test pulse is presented on a prolonged background pedestal, it is strongly masked if presented at the start of the pedestal, and is only weakly masked if presented 200 ms after the start. This has been suggested to occur due to contrast gain mechanisms that reduce the representation of the pedestal and therefore reduce its masking effects. We show here that the effect is due to the large transient in contrast that accompanies the onset of the pedestal. We find similar masking at pedestal offset, when the pedestal is continually flickered, or when pedestal and test have a high spatial frequency. These results were all predicted on the basis of sustained and transient channels. PMID- 11412881 TI - Refractive state and accommodation in the eyes of free-swimming versus restrained juvenile lemon sharks (Negaprion brevirostris). AB - Optical measurements of the refractive state of the eyes of various shark species typically have depicted sharks as hyperopic (far-sighted) with little evidence of accommodation (i.e. the ability to change focus for visualizing objects at different distances from the eye). In this study, we used infrared video retinoscopy to measure the refractive state in juvenile lemon sharks (Negaprion brevirostris). This technique allows dynamic measurement of refractive state in free-swimming animals as they pass by an aquarium window. We found that unrestrained lemon sharks are focused emmetropically relative to a 1-m distant photorefractor for the lateral visual field. However, when restrained either right side up or upside down (the latter inducing tonic immobility), the sharks become increasingly hyperopic, an artifact also reported in some other vertebrates. In addition, unrestrained lemon sharks display small amplitude accommodative excursions. Thus, refractive state measurements on restrained sharks in general may not reflect the natural, resting state of the shark eye, but rather, an induced hyperopia and lack of accommodative function. Such an artifact may be present in other vertebrate species, underscoring the need to obtain measurements of refractive state in unrestrained animals. PMID- 11412882 TI - Temporal integration of optic flow, measured by contrast and coherence thresholds. AB - We measured, as a function of exposure duration, contrast sensitivity and coherence sensitivity for discerning the direction of motion of random dot patterns moving in circular, radial or translational directions. Contrast sensitivity for these patterns increased linearly with exposure duration, up to about 200-300 ms, consistent with previous estimates of temporal summation of early motion units. Coherence sensitivity, however, showed much longer summation periods, about 3 s. When the stimulus was embedded within 10 s of noise, sensitivity increased with duration up to 2-3 s, approximately linearly, as expected from an ideal integrator. When presented without the noise period, sensitivity also increased, but in a different way. For radial and circular motion the increase tended towards the theoretically predicted square root relationship for the same duration as that found with the embedded noise (about 3 s). For translation, however, the curve was steeper than the theoretical prediction (nearly linear), and the summation estimates of around 1000 ms. When the duration of the target was constant at 200 ms, but that of the flanking noise varied, sensitivity decreased with total duration over a similar interval. We interpret our results to reflect at least two stages of analysis, a threshold limited early stage of local-motion analysis, with a time constant of 200-300 ms, and a later global-motion integration stage with a much longer time constant, around 3000 ms. There may also exist an intermediate stage, with an integration time of around 1000 ms. PMID- 11412883 TI - Broad direction bandwidths for complex motion mechanisms. AB - Growing evidence from psychophysics and single-unit recordings suggests specialised mechanisms in the primate visual system for the detection of complex motion patterns such as expansion and rotation. Here we used a subthreshold summation technique to determine the direction tuning functions of the detecting mechanisms. We measured thresholds for discriminating noise and signal+noise for pairs of superimposed complex motion patterns (signal A and B) carried by random dot stimuli in a circular 5 degrees field. For expansion, rotation, deformation and translation we found broad tuning functions approximated by cos(d), where d is the difference in dot directions for signal A and B. These data were well described by models in which either: (a) cardinal mechanisms had direction bandwidths (half-widths) of around 60 degrees; or (b) the number of mechanisms was increased and their half-width was reduced to about 40 degrees. When d=180 degrees we found summation to be greater than probability summation for expansion, rotation and translation, consistent with the idea that mechanisms for these stimuli are constructed from subunits responsive to relative motion. For deformation, however, we found sensitivity declined when d=180 degrees, suggesting antagonistic input from directional subunits in the deformation mechanism. This is a necessary property for a mechanism whose job is to extract the deformation component from the optic flow field. PMID- 11412884 TI - Contextual influences on orientation discrimination: binding local and global cues. AB - We sought to determine how local and global features within an image interact by examining whether orientation discrimination thresholds could be modified by contextual information. In particular, we investigated how local orientation signals within an image are pooled together, and whether this pooling process is dependent on the global orientation content present in the image. We find that observers' orientation judgments depend on surround contextual information, with performance being optimal when the center and surround stimuli are clearly distinct. In cases where the center and surround were not clearly segregated, we report two sets of results. If there was an ambiguity regarding the perception of a global structure (i.e. a small mismatch between local cues), observers' performance was impaired. If there was no mismatch and local and global cues were consistent with the perception of a single surface, observers performed as well as in the distinct surfaces case. Although some of our results can be largely accounted for by interactions between differently oriented filters, other aspects are more difficult to reconcile with this explanation. We suggest that low level filtering constrains observers' performance, and that influences arising from image segmentation modify how local orientation signals are pooled together. PMID- 11412885 TI - Size tuning in the absence of spatial frequency tuning in object recognition. AB - How do we attend to objects at a variety of sizes as we view our visual world? Because of an advantage in identification of lowpass over highpass filtered patterns, as well as large over small images, a number of theorists have assumed that size-independent recognition is achieved by spatial frequency (SF) based coarse-to-fine tuning. We found that the advantage of large sizes or low SFs was lost when participants attempted to identify a target object (specified verbally) somewhere in the middle of a sequence of 40 images of objects, each shown for only 72 ms, as long as the target and distractors were the same size or spatial frequency (unfiltered or low or high bandpassed). When targets were of a different size or scale than the distractors, a marked advantage (pop out) was observed for large (unfiltered) and low SF targets against small (unfiltered) and high SF distractors, respectively, and a marked decrement for the complementary conditions. Importantly, this pattern of results for large and small images was unaffected by holding absolute or relative SF content constant over the different sizes and it could not be explained by simple luminance- or contrast-based pattern masking. These results suggest that size/scale tuning in object recognition was accomplished over the first several images (<576 ms) in the sequence and that the size tuning was implemented by a mechanism sensitive to spatial extent rather than to variations in spatial frequency. PMID- 11412886 TI - Countermanding saccades with auditory stop signals: testing the race model. AB - In a stop signal paradigm to investigate the control of human saccades subjects were instructed to make a saccade to a visual target appearing suddenly l5 degrees to the left or to the right of the fixation point. In 25% of the trials an auditory stop signal was presented after a variable delay that required the subject to inhibit the saccade. The stop signal was presented randomly at the target position, at the opposite side, or at fixation. Using different estimation techniques the average time needed to inhibit a saccade (stop signal processing time, or SSPT) was estimated on the basis of the race model. The SSPT estimates ranging from 50 to 100 ms (depending on subject) are shorter than those from previous studies with visual stop signals. Position of the auditory stop signal did not show an effect on countermanding effectiveness. We found saccadic response times consistent with the race model predictions for two subjects, while a third subject showed small but consistent violations. Moreover, all subjects showed a tendency towards hypometric saccades for responses that could not be inhibited. These findings are discussed with respect to recent neurophysiological results. PMID- 11412887 TI - The relative sensitivities of sensory and motor fusion to small binocular disparities. AB - Horizontal binocular disparity is the fundamental stimulus for both fusional vergence and stereopsis, but whether common disparity-sensitive mechanisms are involved in both responses is unknown. To determine whether the sensitivities of motor and sensory fusion are interdependent, we studied vergence eye movements and depth discrimination, using stimuli with haplopic binocular disparities, in subjects with normal stereopsis and in subjects with mild to severe stereoanomalies. Our results showed that the subjects' disparity discrimination functions varied from nearly perfect discrimination to chance performance for all of the experimental stimuli. Their sensory functions did not necessarily predict the shape of their motor fusion functions, but in most cases were correlated with the subject's fixation disparities. The results support the conclusion that the stereoanomalies and vergence anomalies that previously have been described for coarse binocular disparities also extend to the small, haplopic binocular disparities. The independence of the response properties of sensory and motor fusion suggests that neural pathways for sensory and motor fusion separate after the initial disparity-selective mechanisms in primary visual cortex. PMID- 11412888 TI - What defines a view? AB - At a given instant we see only visible surfaces, not an object's complete 3D appearance. Thus, objects may be represented as discrete 'views' showing only those features visible from a limited range of viewpoints. We address how to define a view using Koenderink's (Koenderink & Van Doorn, Biol. Cybernet. 32 (1979) 211.) geometric method for enumerating complete sets of stable views as aspect graphs. Using objects with known aspect graphs, five experiments examined whether the perception of orientation is sensitive to the qualitative features that define aspect graphs. Highest sensitivity to viewpoint changes was observed at locations where the theory predicts qualitative transitions, although some transitions did not affect performance. Hypotheses about why humans ignore some transitions offer insights into mechanisms for object representation. PMID- 11412889 TI - Visual evoked potentials elicited by chromatic motion onset. AB - Visually Evoked Potentials (VEPs) were recorded in response to the onset of chromatic and luminance motion gratings of 1 cpd and luminance 40 cd m(-2) subtending a 7 degrees field. At slow speeds (< or =2 cycles s(-1)) the motion onset response exhibits a clear amplitude minimum at isoluminance. Over the Michelson contrast range tested (0.05-0.75) the chromatic response at 2 cycles s( 1) possesses a linear response function compared to the saturating function of the luminance response and the contrast dependency of the former is a factor of 5 6 times greater than for the latter. These differences are suggestive of different neural substrates for the chromatic and luminance motion VEPs at slow speeds. At 10 cycles s(-1) the chromatic motion onset VEP exhibits no amplitude minimum at isoluminance and becomes more like its luminance counterpart in terms of its saturating contrast response function. Furthermore, the contrast dependency of the chromatic and luminance responses differs by only a factor of 1.6 at this faster rate. These findings are consistent with the idea of separate motion mechanisms that operate at fast and slow speeds, the latter having separate channels for colour and luminance motion. PMID- 11412891 TI - The biochemistry of hypo- and hyperlipidemic fatty acid derivatives: metabolism and metabolic effects. AB - A selection of amphipatic hyper- and hypolipidemic fatty acid derivatives (fibrates, thia- and branched chain fatty acids) are reviewed. They are probably all ligands for the peroxisome proliferation activation receptor (PPARalpha) which has a low selectivity for its ligands. These compounds give hyper- or hypolipidemic responses depending on their ability to inhibit or stimulate mitochondrial fatty acid oxidation in the liver. The hypolipidemic response is explained by the following metabolic effects: Lipoprotein lipase is induced in liver where it is normally not expressed. Apolipoprotein CIII is downregulated. These two effects in liver lead to a facilitated (re)uptake of chylomicrons and VLDL, thus creating a direct transport of fatty acids from the gut to the liver. Fatty acid metabolizing enzymes in the liver (CPT-I and II, peroxisomal and mitochondrial beta-oxidation enzymes, enzymes of ketogenesis, and omega-oxidation enzymes) are induced and create an increased capacity for fatty acid oxidation. The increased oxidation of fatty acids "drains" fatty acids from the body, reduces VLDL formation, and ultimately explains the antiadiposity and improved insulin sensitivity observed after administration of peroxisome proliferators. PMID- 11412892 TI - The roles of PPARs in adipocyte differentiation. AB - Adipose tissue development takes place primarily around birth but adipose cell number can increase throughout life in response to nutritional changes. At the molecular level, adipogenesis is the result of transcriptional remodeling that leads to activation of a considerable number of genes. Several transcription factors act cooperatively and sequentially in this process. This article attempts to review the roles of peroxisome proliferator-activated receptors gamma and delta in the control of preadipocyte proliferation and differentiation during adipose tissue development or during the adaptive response of adipose tissue mass to high-fat feeding. PMID- 11412893 TI - The biologically active isomers of conjugated linoleic acid. AB - Numerous physiological effects are attributed to conjugated linoleic acid (CLA). The purpose of this presentation is to consider these effects with respect to the cis-9,trans-11 and trans-10,cis-12 CLA isomers. We review previously published data and present new findings that relate to underlying biochemical mechanisms of action. Both isomers are natural products. The cis-9,trans-11 isomer is the principal dietary form of CLA, but the concentrations of this isomer and the trans-10,cis-12 isomer in dairy products or beef vary depending on the diet fed to cows or steers, respectively. The trans-10,cis-12 CLA isomer exerts specific effects on adipocytes, in particular reducing the uptake of lipid by inhibiting the activities of lipoprotein lipase and stearoyl-CoA desaturase. The trans 10,cis-12 CLA isomer also affects lipid metabolism in cultured Hep-G2 human liver cells, whereas both the cis-9,trans-11 and trans-10,cis-12 CLA isomers appear to be active in inhibiting carcinogenesis in animal models. We present new findings indicating that the cis-9,trans-11 CLA isomer enhances growth and probably feed efficiency in young rodents. Accordingly, the effects of CLA on body composition (induced by trans-10,cis-12 CLA) and growth/feed efficiency (induced by cis 9,trans-11 CLA) appear to be due to separate biochemical mechanisms. We also show that a 19-carbon CLA cognate (conjugated nonadecadienoic acid, CNA) inhibits lipoprotein lipase activity as effectively as CLA in cultured 3T3-L1 adipocytes. Presumably, CNA is metabolized differently than the 18-carbon CLA isomers, so this finding indicates direct activity of the administered compound as opposed to acting via a metabolite. PMID- 11412894 TI - Do sterols reduce proton and sodium leaks through lipid bilayers? AB - Proton and/or sodium electrochemical gradients are critical to energy handling at the plasma membranes of all living cells. Sodium gradients are used for animal plasma membranes, all other living organisms use proton gradients. These chemical and electrical gradients are either created by a cation pumping ATPase or are created by photons or redox, used to make ATP. It has been established that both hydrogen and sodium ions leak through lipid bilayers at approximately the same rate at the concentration they occur in living organisms. Although the gradients are achieved by pumping the cations out of the cell, the plasma membrane potential enhances the leakage rate of these cations into the cell because of the orientation of the potential. This review proposes that cells use certain lipids to inhibit cation leakage through the membrane bilayers. It assumes that Na(+) leaks through the bilayer by a defect mechanism. For Na(+) leakage in animal plasma membranes, the evidence suggests that cholesterol is a key inhibitor of Na(+) leakage. Here I put forth a novel mechanism for proton leakage through lipid bilayers. The mechanism assumes water forms protonated and deprotonated clusters in the lipid bilayer. The model suggests how two features of lipid structures may inhibit H(+) leakage. One feature is the fused ring structure of sterols, hopanoids and tetrahymenol which extrude water and therefore clusters from the bilayer. The second feature is lipid structures that crowd the center of the bilayer with hydrocarbon. This can be accomplished either by separating the two monolayers with hydrocarbons such as isoprenes or isopranes in the bilayer's cleavage plane or by branching the lipid chains in the center of the bilayers with hydrocarbon. The natural distribution of lipids that contain these features are examined. Data in the literature shows that plasma membranes exposed to extreme concentrations of cations are particularly rich in the lipids containing the predicted qualities. Prokaryote plasma membranes that reside in extreme acids (acidophiles) contain both hopanoids and iso/anteiso- terminal lipid branching. Plasma membranes that reside in extreme base (alkaliphiles) contain both squalene and iso/anteiso- lipids. The mole fraction of squalene in alkaliphile bilayers increases, as they are cultured at higher pH. In eukaryotes, cation leak inhibition is here attributed to sterols and certain isoprenes, dolichol for lysosomes and peroxysomes, ubiquinone for these in addition to mitochondrion, and plastoquinone for the chloroplast. Phytosterols differ from cholesterol because they contain methyl and ethyl branches on the side chain. The proposal provides a structure-function rationale for distinguishing the structures of the phytosterols as inhibitors of proton leaks from that of cholesterol which is proposed to inhibit leaks of Na(+). The most extensively studied of sterols, cholesterol, occurs only in animal cells where there is a sodium gradient across the plasma membrane. In mammals, nearly 100 proteins participate in cholesterol's biosynthetic and degradation pathway, its regulatory mechanisms and cell-delivery system. Although a fat, cholesterol yields no energy on degradation. Experiments have shown that it reduces Na(+) and K(+) leakage through lipid bilayers to approximately one third of bilayers that lack the sterol. If sterols significantly inhibit cation leakage through the lipids of the plasma membrane, then the general role of all sterols is to save metabolic ATP energy, which is the penalty for cation leaks into the cytosol. The regulation of cholesterol's appearance in the plasma membrane and the evolution of sterols is discussed in light of this proposed role. PMID- 11412895 TI - Perinatal expression of heat shock proteins HSC 70 and HSP 70 in neural and non neural tissues of the piglet. AB - Stress of different kinds during early perinatal life can result in severe consequences for further development. To determine possible involvement of heat shock proteins in brain development, the expression of HSC 70 and HSP 70 was determined in brain regions (cerebellum, cortex, hippocampus, hypothalamus and striatum) and non neural tissues (liver, lungs and kidneys) at birth and during early development of the piglet. In brain regions, HSC 70 expression was decreased during the few hours following birth. With the exception of cortex, hippocampus and kidney where a decrease of expression was observed, HSP 70 did not show significant changes during early development. These results are discussed in terms of using the piglet model of development to study the effect of different kinds of stress like hypoxia or temperature changes on brain development. PMID- 11412896 TI - Developmental expression of prolactin releasing peptide in the rat brain: localization of messenger ribonucleic acid and immunoreactive neurons. AB - Prolactin releasing peptide (PrRP) was recently identified as the stimulator of prolactin release from the anterior pituitary. PrRP mRNA is expressed in the medulla oblongata and the hypothalamus in the rat brain. The fibers containing PrRP are widely distributed in the brain, therefore, it was postulated that PrRP may act as a neurotransmitter or neuromodulator as well as an endocrine substance. To clarify the developmental changes in the expression of PrRP during brain development, we examined PrRP in rat fetuses and neonates using in situ hybridization and immunohistochemistry. The PrRP mRNA was expressed in the nucleus of the solitary tract (NTS) at embryonic day 18 (E18) and in the ventral and lateral reticular nucleus (VLRN) of the caudal medulla oblongata at E20. The PrRP mRNA in the hypothalamus was first expressed at postnatal day 13 (P13). Reverse transcription-polymerase chain reaction analysis (RT-PCR) for PrRP revealed that PCR product, a 268 bp band, was detected from either E18 in the medulla or P13 in the hypothalamus. Immunodetection with monoclonal antibody against prepro-PrRP revealed intensive staining of cells in the NTS at E18, in the VLRN at E20 or in the dorsomedial hypothalamus at P13. Immunohistochemistry using monoclonal antibody against mature PrRP at P6 showed PrRP fibers to be distributed in the paraventricular hypothalamic nucleus, periventricular hypothalamic nucleus, medial preoptic area, basolateral amygdaloid nucleus, dorsomedial hypothalamus, ventromedial hypothalamus, periventricular nucleus of the thalamus and bed nucleus of the stria terminalis as previously shown in the adult rat. PrRP fibers were also found in the optic chiasm, dorsal endopiriform nucleus, cingulum, intermediate reticular nucleus, and caudal ventrolateral reticular nucleus at P6 and P9. However, PrRP fibers were never found in the above regions in the adult animal. These findings suggest that PrRP fibers originating in the medulla oblongata have been widely distributed in the rat brain during the early postnatal day and PrRP may play various roles in the brain development. PMID- 11412897 TI - Developmental characteristics of epileptiform activity in immature rat neocortex: a comparison of four in vitro seizure models. AB - New-onset seizures and epilepsy have a relatively high incidence in infants and children. A leading hypothesis to explain an increased seizure susceptibility of the immature nervous system involves ontogenetic changes in different neurotransmitter systems, such as specific glutamate and GABA receptors. However, few studies have directly tested this hypothesis in a systematic fashion, especially in neocortical structures, where seizures in pediatric patients frequently arise. The present study investigated developmental changes in epileptiform activity in rat neocortical slices from four age groups (postnatal days P4--7, P13--16, P23--26, P41--47) due to four pharmacological conditions (4 aminopyridine, low magnesium, picrotoxin, CGP-35348) that differentially modulate glutamate and GABA systems. A characteristic age-dependence of the incidence of epileptiform activity was observed. In all pharmacological conditions, no epileptiform activity occurred in neocortical slices from P4--7 rats. Interictal discharges, ictal events, and spreading depression had a maximal incidence at P13 -16 and decreased progressively in later age groups. 4-Aminopyridine, low magnesium, and picrotoxin induced all types of epileptiform activity with a similar age-dependent pattern, despite minor differences in quantitative characteristics of epileptiform activity between these three conditions. The GABA(B) antagonist, CGP-35348, did not elicit epileptiform activity in any age group, but could potentiate synaptic potentials. These findings establish that isolated neocortical tissue intrinsically displays ontogenetic changes in seizure susceptibility independent of systemic factors. The similar age-dependent patterns of epileptiform activity with multiple drugs support a concept of global developmental changes in excitability not specifically linked to any particular neurotransmitter system. PMID- 11412898 TI - Ontogenesis of neurons immunoreactive for nitric oxide synthase in rat forebrain and midbrain. AB - We studied the immunohistochemical localization of neuronal nitric oxide synthase (nNOS) in the developing rat brain on embryonic days 13 (E13), 15 (E15), 17 (E17) and 19 (E19) and postnatal days 0 (P0), 7 (P7) and 14 (P14). A few neurons positive for nNOS were first detected at E15 in the hypothalamus and pons. At E17, many positive cells became detectable in the thalamus. At E19, the positive cells in these three regions were rapidly increased in number, and a few positive neurons were also observed in such regions as the cerebral cortex and striatum. Positive cells in the hypothalamus tended to locate ventrolaterally. Positive neurons, stained very intensely as in adult rats, were seen in the pedunculopontine tegmental nucleus, laterodorsal tegmental nucleus and parafascicular nucleus. Two weeks after birth, positive neurons of larger somata with many processes were distributed widely in the cerebral cortex and hippocampus. The present study indicates that, in the forebrain and midbrain, the distribution pattern of nNOS-containing neurons is fundamentally completed by E19. PMID- 11412899 TI - MK801 decreases glutamate release and oxidative metabolism during hypoglycemic coma in piglets. AB - Hypoglycemic coma increases extracellular excitatory amino acids, which mediate hypoglycemic neuronal degeneration. Cerebral oxygen consumption increases during hypoglycemic coma in piglets. We tested the hypothesis that the NMDA-receptor antagonist dizocilpine (MK801) attenuates the increase in cerebral oxygen consumption during hypoglycemia. We measured EEG, cerebral blood flow (CBF), cerebral oxygen consumption (CMRO(2)) and cortical microdialysate levels of glutamate, aspartate and glycine in pentobarbital-anesthetized piglets during 60 min of insulin-induced hypoglycemic coma. NMDA-receptor distribution was measured by autoradiography. MK801 (0.75 mg/kg i.v.) was given within 5 min after onset of isoelectric EEG. Saline- and MK801-treated normoglycemic control animals were also studied. Brain temperature was maintained at 38.5+/-0.5 degrees C. MK801 prevented the 5--10-fold increase in glutamate and aspartate occurring in saline treated hypoglycemic animals, and attenuated the increase in CMRO(2). Increases in CBF of 200--400% during hypoglycemic coma were not affected by MK801. MK801 did not alter CBF, CMRO(2) or microdialysate amino acid levels in normoglycemic control animals. Parietal cortex corresponding to microdialysis sites was highly enriched in NMDA receptors, and the density and distribution overall of NMDA receptor binding sites were comparable to that reported in other species. We conclude that NMDA receptor activation plays a central role in hypoglycemia induced glutamate release, and contributes to increased cerebral oxygen consumption. Neuroprotective effects of MK801 during hypoglycemia in piglets may involve inhibitory effects on glutamate release and oxidative metabolism. PMID- 11412900 TI - Development of retino-recipient projection neurons in the optic tectum of the chicken. AB - The dendritic development of a well-characterized retino-recipient neuronal type in the chicken optic tectum has been traced with intracellular labeling. Normal dendritic development can be divided into three phases: extension, differentiation and pruning. During the first phase, cells extend their dendrites, generate large dendritic fields and position their distal endings in a certain retino-recipient tectal layer. In the second phase, these dendritic endings arborize into characteristic bottlebrush-like structures, while the overall morphology of the neurons remains unaltered. After hatching, the number and width of the bottlebrush endings are reduced. The findings are discussed with respect to the innervation of the optic tectum by retinal afferents and possible guidance mechanisms for synapse formation in this system. PMID- 11412901 TI - Potential involvement of S100B in the protective effects of a serotonin-1a agonist on ethanol-treated astrocytes. AB - Previously, this laboratory found that the offspring of rats that consumed ethanol on a chronic basis prior to parturition exhibited a significant reduction in serotonin (5-HT) neurons and in astrocytes proximal to these neurons. This laboratory also showed that maternal treatment with a 5-HT(1A) agonist during the latter part of gestation prevented the reduction of 5-HT neurons and most of the astrocyte abnormalities. The present in vitro studies extended our prior in vivo work by examining the potential involvement of S100B with the protective effects of a 5-HT(1A) agonist, i.e., buspirone, on astrocytes. Astrocyte cultures were either maintained in chemically defined media in the presence and absence of ethanol and buspirone or in conditioned media that was generated by ethanol- and buspirone-treated astrocytes. A mouse monoclonal antibody to S100B was used to assess the potential involvement of S100B with the protective effects of buspirone. Additional in vitro studies measured the direct effects of S100B and ethanol on astrocyte proliferation. These investigations demonstrate that in vitro ethanol exposure reduces the number of astrocytes, and that treatment with the 5-HT(1A) agonist buspirone prevents the ethanol-associated reduction in astrocyte number. The protective effects of buspirone appear to be mediated by factors that are secreted by astrocytes; such factors likely include S100B. In addition, added S100B prevents an ethanol-associated reduction in [(3)H] thymidine incorporation into proliferating astrocytes. PMID- 11412902 TI - Localization of estrogen receptor alpha (ER alpha) in pyramidal neurons of the developing rat hippocampus. AB - During development, estrogen has a variety of effects on morphological, biochemical and electrophysiological properties of hippocampal neurons. Correspondingly, estrogen receptor (ER) binding and mRNA increase transiently in the developing hippocampus. In this study, we used immunocytochemistry to determine the localization of the ER alpha subtype in the developing rat hippocampus. Nuclear staining was present in pyramidal cells and some interneurons of the CA1 and CA3 regions of the developing rat hippocampus. Little or no immunoreactivity was observed in postnatal day (P)0 animals (day of birth=P0), however, beginning on P4, ER alpha-immunoreactivity (ER alpha-ir) was visible and reached maximal levels by P10. These levels subsequently declined to low levels so that by P15, levels approximated those of adult females. Western blot analysis confirmed that this antibody recognized a 67 kDa protein, characteristic of the full-length ER alpha protein, in the hippocampus and pituitary. Furthermore, most of the ER alpha-immunopositive cells in the hippocampus were located in the pyramidal cell layer, and did not co-localize appreciably with gamma-aminobutyric acid (GABA) at any age examined. We conclude, based on the immunocytochemical localization of ER alpha, that the effects of estrogen on biochemistry and morphology of the developing hippocampus may be direct through the ER alpha subtype in hippocampal pyramidal cells. PMID- 11412903 TI - Infusion of the metabotropic receptor agonist, DCG-IV, into the main olfactory bulb induces olfactory preference learning in rat pups. AB - DCG-IV, a type 2 metabotropic glutamate receptor (mGluR2) agonist, was infused into the main olfactory bulb of 1-week-old pups exposed to peppermint odor. A preference for peppermint was demonstrated 24 h later. The data support the proposal that disinhibition at dendrodendritic synapses between granule cells and mitral cells is a critical component of olfactory memory formation in the rat pup olfactory bulb as well as in the accessory olfactory bulb of adult rodents. PMID- 11412904 TI - In vivo PC3 overexpression by retroviral vector affects cell differentiation of rat cortical precursors. AB - The PC3 gene is a marker of dividing neuroepithelial (NE) cells. We transduced single cortical precursors of the ventricular zone (VZ) with a PC3-carrying retroviral vector at E16 stage, and analysed the effects of transgene expression on their progeny in 3-week-old animals. Unlike control-transduced cells, all viable PC3-transduced cells remained close to the ventricle and displayed a round shaped, undifferentiated morphology. PMID- 11412905 TI - Bid regulation of neuronal apoptosis. AB - Bid is a BH3 domain only pro-apoptotic member of the Bcl-2 family which interacts with Bax to regulate apoptosis. Bax-deficient embryos show decreased neuronal programmed cell death in vivo and resistance to cytosine arabinoside (AraC) induced neuronal apoptosis in vitro. In this report, we demonstrate that Bid deficient embryos show no neurodevelopmental abnormalities, and Bid-deficiency has no effect on the in vitro apoptotic response of either telencephalic neural precursor cells or neurons to AraC-induced death. We conclude that bid does not play an essential role in either naturally occurring or genotoxin-induced neuronal cell death. PMID- 11412906 TI - The development of vesicular acetylcholine transporter immunoreactivity in the hindlimbs of the opossum Monodelphis domestica. AB - Vesicular acetylcholine transporter (VAChT) was revealed immunohistochemically in light microscopy on hindlimb sections of developing opossums, Monodelphis domestica. In the immobile hindlimbs of the newborn, which comprise cartilaginous bones and loose, unstriated myofibers, scant immunolabeled nerve segments and small spherical terminals, presumably growth cones or immature neuromuscular junctions, are found in the muscle tissue of the thigh, leg and proximal foot, decreasing in number and size proximodistally. When the hindlimbs start moving at 1 week, terminals are more numerous and larger, still decreasing proximodistally, and occur in the newly formed interosseous foot muscles. At 4 weeks, when the hindlimbs start supporting weight and quadrupedal locomotion appears, terminals are more numerous, flattened and in comparable size and density in thigh, leg and foot muscles. By 7 weeks, large and completely flat terminals are observed in groups of 3 to 4 at regular intervals along muscle fibers. VAChT expression develops largely postnatally in the opossum hindlimbs, along a proximodistal gradient that parallels somatic and reflex development. PMID- 11412908 TI - Raising antibodies against OprD, an outer membrane protein of Pseudomonas aeruginosa using translational fusions to MalE. AB - OprD is an outer membrane porin of Pseudomonas aeruginosa that mediates uptake of basic amino acids, peptides as well as carbapenem antibiotics. Polyclonal antibodies were raised against the OprD porin by creating protein fusions between the Escherichia coli maltose binding protein and four OprD fragments. These were expressed in E. coli and shown to be exported to the periplasm. The fusion proteins were purified by amylose affinity chromatography and used to immunize rabbits intramuscularly. We established that MalE fusions to OprD fragments retain maltose and amylose binding activities in vivo and in vitro, confirming proper folding of the MalE domain of hybrid proteins. Furthermore, we demonstrate that this strategy can be used to obtain specific antibodies against bacterial outer membrane proteins (OMPs). PMID- 11412909 TI - Uniaxial compression measurement device for investigation of the mechanical stability of biofilms. AB - The mechanical stability of biofilms is important for biotechnology, as sloughing of the biomass due to mechanical failure of the biofilm matrix can lead to severe interferences with biofilm processes. In cases of biofouling, biofilms have to be removed, in which case their mechanical stability must be overcome. The apparent modulus of elasticity and the yield strength as obtained from uniaxial compression experiments can be taken as parameters indicative for the mechanical stability of a biofilm. A film rheometer is presented which allows for the determination of these quantities, using model biofilms of Pseudomonas aeruginosa grown on membrane filters. The compressive stress-strain behaviour up to the point of failure is recorded at a compression speed of 1 microm s(-1). In accordance with the stress-strain curve, the investigated biofilm can be described as viscoelastic material, which demonstrates plastic flow properties. The extracellular polymeric substances (EPS), which keep biofilms together, form a temporary network of fluctuating junction points. Above the yield point, the gel structure fails and the system behaves as a highly viscous fluid. The apparent modulus of elasticity and the yield point are considered to be useful parameters for characterizing the mechanical properties of biofilms. PMID- 11412910 TI - Detection of respiratory enzyme activity in Giardia cysts and Cryptosporidium oocysts using redox dyes and immunofluorescence techniques. AB - The fluorescent redox dye 5-cyano-2,3-ditolyl tetrazolium chloride (CTC), combined with fluorescein-labeled antibodies, was tested for the simultaneous detection of the respiratory electron transport system (ETS) activity and enumeration of Giardia cysts and Cryptosporidium oocysts by spectral microfluorometry and epifluorescence microscopy. The reduction of CTC and p iodonitrotetrazolium violet (INT), a non-fluorescent redox dye, was compared with propidium iodide (PI) and fluorescein diacetate (FDA) for the measurements of Giardia cyst viability over time. According to the PI and FDA staining techniques, nearly 60% of the cysts tested viable at the beginning of the observations; after 21 days their viability decreased to 5%. The redox dyes indicated that approximately 4-10% of the cysts were metabolically active 48 h after they were shed, followed by a decline in enzyme activity to near undetectable levels after 4 days. Spectral analysis on individual cysts indicated that the fluorescence emission of the reduced CTC and the fluorescein-labeled antibodies is distinctive for each compound and suitable for their simultaneous determination by microphotometry, flow cytometry and epifluorescence microscopy. The fluorescence signal remained without alteration when the cysts were transferred onto microscope slides coated with an optical embedding medium and stored at -20 degrees C. The fluorescence intensity of the reduced CTC, when properly standardized, can provide quantitative measurements of ETS activity of the cysts. This is the first report of a method to determine enzyme redox activity on intact cysts applicable to water, laboratory and animal samples. PMID- 11412911 TI - Improved detection and differentiation of mycobacteria with combination of Mycobacterium Growth Indicator Tube and Roche COBAS AMPLICOR System in conjunction with Duplex PCR. AB - In this study, a combination of liquid and solid media (current "gold standard" for culture) with combinations of liquid media (Mycobacteria Growth Indicator Tube (MGIT)) plus a commercial amplification system (Roche COBAS AMPLICOR System (CAS)), and solid media (Ogawa) plus CAS for detection of Mycobacterium tuberculosis were compared. In addition, the ability of the MGIT to recover mycobacteria from various clinical samples was compared with the abilities of egg based Ogawa medium using equal volume of samples and a high concentration (6%) of NaOH for decontamination. A total of 705 specimens (395 respiratory and 310 extrapulmonary) that were collected from 554 patients were tested in parallel with three assays. The results of MGIT and Ogawa were evaluated with the "gold standard" (combination of culture and clinical data) and those of CAS were evaluated with extended gold standard including treated tuberculosis. A total of 130 mycobacterial infections (M. tuberculosis, n=122; mycobacterium other than tuberculosis (MOTT), n=8) were detected. The differentiation of M. tuberculosis and MOTT was successfully accomplished using duplex PCR. The overall sensitivity of the MGIT, Ogawa, and CAS for M. tuberculosis was 89.9%, 73.9%, and 79.9%, respectively. For the MOTT, the corresponding values for the MGIT and Ogawa medium were 100% and 12.5%, respectively. The mean detection time for M. tuberculosis was 22 days using MGIT and 32 days when using the Ogawa medium. The specificity of CAS was 98.4%, with an inhibition rate of 1.4%. A combination of MGIT plus CAS detected 97.5% of all M. tuberculosis infections (compared with MGIT plus Ogawa, 91.8%, P<0.05; compared with Ogawa plus CAS, 87.7%. P<0.01). Our results indicate that a combination of MGIT plus a Roche CAS in conjunction with duplex PCR, would be quite useful in clinical laboratories for both rapid detection and differentiation of M. tuberculosis and MOTT. PMID- 11412912 TI - Microbial community assessment in oil-impacted salt marsh sediment microcosms by traditional and nucleic acid-based indices. AB - The effect of oil amendment in salt marsh sediment microcosms was examined by most probable number (MPN), DNA-hybridization with domain-specific oligonucleotide probes and whole community 16S rDNA-hybridizations. Gas chromatography (GC/MS) analysis of oil residues in sediments from microcosms after 3 months of operation showed that the quantity of petroleum hydrocarbons was lower in microcosms amended with oil compared to microcosms amended with oil+plant detritus. Bacterial numbers (total-MPN) increased in all experimental microcosms (amended with plant detritus, oil, and oil+plant detritus). In comparison to the intact sediment, the proportions of oil-degrading bacteria increased >100-fold in the oil amended microcosm and >10-fold in the plant detritus and the oil+plant detritus amended microcosms. DNA-hybridizations with Bacteria, Archaea and Eukarya oligonucleotide probes indicated few changes in the petroleum contaminated sediment community profile. In contrast, rDNA hybridizations indicated that the bacterial community profile of the oil-impacted sediments, after 1 month of exposure, was significantly different from the control sediment. PMID- 11412913 TI - Bacteria in gel probes: comparison of the activity of immobilized sulfate reducing bacteria with in situ sulfate reduction in a wetland sediment. AB - A novel method was used to examine the microbial ecology of iron-rich wetland sediments receiving neutral-pH coal mine drainage. Gel probes inserted into the sediments allowed analysis of the distribution and activity of bacterial sulfate reduction (BSR). A mixed population of sulfate-reducing bacteria enriched from anoxic wetland sediments was immobilized in low temperature-gelling agarose held in grooved rods or probes. The probes were inserted vertically into sediments and were allowed to incubate in situ for 48 h. After their retrieval, the gels were sectioned and analyzed for residual BSR activity and were compared to in situ BSR rates and chemical porewater profiles. The depth distribution of residual BSR activity in the immobilized cell gel probes differed significantly from the BSR measured in situ. Approximately 51% of the total integrated residual sulfate reduction activity measured in the gel probes occurred between 0 and 7 cm of the upper 20 cm of sediment. In contrast, ca. 99% of the integrated in situ BSR occurred between 7- and 20-cm depth, and only 1% of the total integrated rate occurred between 0- and 7-cm depth. Lactate-enriched bacteria immobilized in the gel may have been atypical of the majority of sulfate-reducing bacteria in the sediment. Agarose-immobilized sulfate-reducing bacteria might also be able to proliferate in the otherwise inhospitable zone of iron reduction, where sulfate and labile carbon compounds for which they are usually outcompeted can diffuse freely into the gel matrix. Gel probes containing particulate iron monosulfide (FeS) indicated that FeS remained stable in sediments at depths greater than 2 to 3 cm below the sediment-water interface, consistent with the shallow penetration of oxygen into surface sediments. PMID- 11412914 TI - Separation of Listeria from cheese and enrichment media using antibody-coated microbeads and centrifugation. AB - An immunoseparation system for the separation of Listeria from enriched cheese samples was developed. The system utilizes polystyrene microbeads (3.8 microm in diameter) coated with covalently bound anti-Listeria genus-specific antibody. The beads were incubated with cheese enriched in half-Fraser broth and the bead bacterial complex was separated by centrifugation at 110 g then spread on selective agar plates. Although cross-reactivity with certain Gram-positive bacteria (Staphylococcus saprophyticus and Arthrobacter sp.) was determined, this had no negative effect on capture effectiveness of the beads to Listeria spp. The minimum density of Listeria cells positively detected by immunoseparation with subsequent plating was 10(0) cfu/ml. The application of the separation method facilitates a reduction in the time of Listeria detection in cheese by 2 days without affecting the sensitivity. PMID- 11412915 TI - Combination of ATP-bioluminescence and PNA probes allows rapid total counts and identification of specific microorganisms in mixed populations. AB - We have combined ATP-dependent bioluminescence with a novel chemiluminescent in situ hybridization (CISH) method using peroxidase-labeled peptide nucleic acid (PNA) probes targeting species-specific rRNA sequences to provide total counts and subsequent identification of specific microorganisms. Both methods are applied to the same membrane filter following a short incubation time and both methods provide results in the form of spots of light that are captured by the MicroStar detection system. Each spot of light represents individual micro colonies detected by either ATP bioluminescence or PNA CISH. This new concept is particularly intended for in process and quality control of non-sterile products to rapidly provide total counts as well as presence/absence of specific indicators and/or pathogens in non-sterile, filterable samples. PMID- 11412916 TI - Reliable use of green fluorescent protein in fluorescent pseudomonads. AB - When fluorescent pseudomonads are cultured on standard solid media under iron limiting conditions, they produce fluorescent, pigmented iron collating agents (siderophores). Siderophores can be readily identified by strong fluorescence seen under UV/blue light. The application of the eukaryotic green fluorescent protein (GFP) as a bacterial marker in microbial ecology is increasingly being used, particularly as it is a powerful method for non-destructive monitoring in situ. As gfp expressing bacteria have to be detected under UV/blue light, the fluorescence of siderophore-producing Pseudomonas spp. masks normal levels of GFP fluorescence when colonies are viewed on standard bacterial agar. Here, we describe a simple but effective way of identifying gfp-expressing Pseudomonas fluorescens using media supplemented with 0.45 mM FeSO(4).7H(2)O. This is of relevance for the screening of insertion libraries and in the application of GFP transposons as promoter probes. PMID- 11412917 TI - Use of Percoll for the infection of cells in vitro with Cryptosporidium parvum oocysts. AB - A method for the infection of non-adherent THP-1 cells and adherent MDBK cells with Cryptosporidium parvum oocysts using isotonic Percoll solutions was developed. Excystation was maximal after 2 h, but toxicity increased with the oocyst/cell ratio and the incubation time. The infection rates did not increase with the oocyst/cell ratio and both cell types were equally parasitized. PMID- 11412918 TI - Formula thinking, formula science and kits. PMID- 11412919 TI - Use of surface-enhanced laser desorption ionization protein chip system to analyze streptococcal exotoxin B activity secreted by Streptococcus pyogenes. AB - Ciphergen surface-enhanced laser desorption ionization (SELDI) protein chip technology was used to analyze the secretion and autoactivation of the Streptococcus pyogenes cysteine protease SpeB. This method allowed rapid identification of both the zymogen form of the protein Mr approximately 41,000 and the fully active enzyme Mr approximately 28,500. SpeB production in culture supernatants was demonstrated to be growth-phase regulated and SpeB positive and negative variants of a blood passaged S. pyogenes isolate could readily be distinguished. In kinetic studies of the autoactivation of the zymogen form of SpeB, the sequential generation of four intermediates was detected before the accumulation of the fully active enzyme. The methods described enabled enhanced speed, use of lower sample volumes and concentrations, and a more complete molecular characterization of SpeB than allowed by existing methods of analysis using SDS-PAGE and Western immunoblotting. PMID- 11412920 TI - Identification of mutans streptococcal species by the PCR products of the dex genes. AB - A pair of polymerase chain reaction (PCR) primers was designed on the basis of the nucleotide sequence homology of dextranase genes (dex) of Streptococcus mutans, S. sobrinus and S. downei. The primer pair amplified a 530-bp DNA fragment on the dex genes of mutans streptococcal species: S. mutans, S. sobrinus, S. downei, S. rattus and S. cricetus. HaeIII digestion of the 530-bp fragments generated species-specific subfragments, which were easily distinguishable from each other by agarose gel electrophoresis. These results suggest that the PCR-amplification of the dex gene followed by the HaeIII digestion is useful for rapid identification of the five species of mutans streptococci. PMID- 11412921 TI - Isolation of human plasma-inducible, growth phase- and temperature-regulated gene fusions in Streptococcus pyogenes using a Tn917-lacZ transposon. AB - Streptococcus pyogenes is capable of causing a variety of human diseases ranging from superficial or deep tissue infections to non-infectious post-streptococcal infection sequelae. In this paper, we report the use of a Tn917-lacZ transposon to isolate random lacZ transcription fusions in the S. pyogenes chromosome. Libraries of random Tn917-lacZ mutants were generated in a representative opacity factor positive strain CS101 (M49) and an opacity factor negative strain 1881 (M1). Several different mutant phenotypes were isolated. These included: temperature-regulated promoters, growth phase/cell density-regulated promoters and a human plasma-inducible promoter. Expression of the temperature-regulated fusions was 5-10-fold higher when grown at 30 degrees C compared to growth at 37 degrees C. The growth phase-regulated fusions were induced 30-fold at late exponential phase and were repressed by a diffusible S. pyogenes factor(s). Expression of the human plasma-inducible fusion was induced 10-15-fold by human plasma or sera, 4-fold by rabbit sera and was repressed by horse and mouse sera. In addition, hemolysin negative and capsule over expression mutants were isolated. These results demonstrate the utility of Tn917-lacZ mutagenesis for the identification of S. pyogenes promoters. PMID- 11412922 TI - Measurements of net fluxes and extracellular changes of H+, Ca2+, K+, and NH4+ in Escherichia coli using ion-selective microelectrodes. AB - This study introduced the use of a non-invasive ion-selective microelectrode (MIFE) technique to study membrane-transport processes in bacteria. Net ion fluxes and changes in the extracellular concentrations of H+, Ca2+, K+ and NH4+ in adherent bacteria, isolated from cultures at different growth stages (exponential, late exponential, and stationary phases), were monitored. With the exception of Ca2+, a significant (P=0.05) difference was found in the magnitude of net fluxes of the ions measured from bacterial cells at different stages of the population growth curve. The magnitude of the H+ response was glucose dependent with maximum changes occurring at the highest concentration. There was a progressive increase in H+ extrusion followed by a gradual return to zero at late stationary phase. Measurements of net ion fluxes crossing the bacterial cytoplasmic membrane, demonstrated here for the first time, may offer insight into underlying mechanisms of ion transport kinetics. Applications of the non invasive ion-selective microelectrode technique in microbiology are discussed. PMID- 11412923 TI - Detection of Escherichia coli O157:H7 by multiplex PCR and their characterization by plasmid profiling, antimicrobial resistance, RAPD and PFGE analyses. AB - Twenty-five and three strains of Escherichia coli O157:H7 were identified from 25 tenderloin beef and three chicken meat burger samples, respectively. The bacteria were recovered using the immunomagnetic separation procedure followed by selective plating on sorbitol MacConkey agar and were identified as E. coli serotype O157:H7 with three primer pairs that amplified fragments of the SLT-I, SLT-II and H7 genes in PCR assays. Susceptibility testing to 14 antibiotics showed that all were resistant to two or more antibiotics tested. Although all 28 strains contained plasmid, there was very little variation in the plasmid sizes observed. The most common plasmid of 60 MDa was detected in all strains. We used DNA fingerprinting by randomly amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) to compare the 28 E. coli O157:H7 strains. At a similarity level of 90%, the results of PFGE after restriction with XbaI separated the E. coli O157:H7 strains into 28 single isolates, whereas RAPD using a single 10-mer oligonucleotides separated the E. coli O157:H7 strains into two clusters and 22 single isolates. These typing methods should aid in the epidemiological clarification of the E. coli O157:H7 in the study area. PMID- 11412924 TI - A novel method for the isolation of motile bacteria using gradient culture systems. AB - Isolation of motile bacteria from stream water samples was achieved by using Lutrol F127 (poloxamer 407) as a gelling agent in culture media. This block copolymer has the property of repeatedly liquefying and solidifying at low and high temperatures, respectively. The ability of motile bacteria to move through liquid-state Lutrol F127 towards a higher nutrient concentration was exploited. After establishment of the nutrient gradient and inoculation, the system was cooled to liquefy the medium and kept liquid to allow motile bacteria to move. Raising the temperature allowed solidification and prevented further movement. Colonies could be easily removed. The proportion of motile isolates (determined by microscopic observation) increased from 42% in the indigenous population to 100% after isolation using the gradient system. PMID- 11412925 TI - Recovery of Rhodococcus biosurfactants using methyl tertiary-butyl ether extraction. AB - In the present study, we proposed methyl tertiary-butyl ether (MTBE) as a solvent for extraction of biosurfactants from Rhodococcus bacterial cultures. After comparison with other well known solvent systems used for biosurfactant extraction, it was found that MTBE was able to extract crude surfactant material with high product recovery (10 g/l), efficiency (critical micelle concentration (CMC), 130-170 mg/l) and good functional surfactant characteristics (surface and interfacial tensions, 29 and 0.9 mN/m, respectively). The isolated surfactant complex contained 10% polar lipids, mostly glycolipids possessing maximal surface activity. Ultrasonic treatment of the extraction mixture increased the proportion of polar lipids in crude extract, resulting in increasing surfactant efficiency. Due to certain characteristics of MTBE, such as relatively low toxicity, biodegradability, ease of downstream recovery, low flammability and explosion safety, the use of this solvent as an extraction agent in industrial scale biosurfactant production is feasible. PMID- 11412926 TI - Evaluation of enhanced haemolysis agar for detection of Listeria spp. and L. monocytogenes from production lines of fresh to cold-smoked fish. AB - Enhanced haemolysis agar (EHA) was compared to the two conventional Listeria isolation agars Oxford and PALCAM for its ability to detect Listeria spp. from production lines of fresh to cold-smoked fish. The ability of EHA for distinguishing L. monocytogenes colonies from other Listeria spp. was also evaluated.A total of 243 fish and environmental samples were analysed. Overall, 42 samples were found to contain Listeria spp. Only 34 samples were positive simultaneously by the three plating media. Two samples considered to be negative by the two conventional agars were found to be positive after isolation on EHA. All three selective agars were shown to be less effective in recovering Listeria spp. after primary enrichment in half-Fraser broth, compared to secondary enrichment in Fraser broth after 24 and 48 h. From 79 Listeria but presumptive negative L. monocytogenes colonies, EHA identified correctly 76 Listeria spp. and presented three false-negative results_three colonies further identified as L. monocytogenes but showing no noticeable haemolysis on EHA. Twenty-three of the thirty-three L. monocytogenes presumptive positive colonies, were confirmed positive and ten were identified as L. seeligeri. Despite its ability of distinguishing L. monocytogenes from the other Listeria spp., unless it is produced as a commercial medium, EHA cannot be an alternative to time-consuming classical identification because the preparation of this medium is both time and labour intensive. PMID- 11412947 TI - Long-term data storage in DNA. AB - This article discusses how DNA might be used to store data. It is argued that, at present, DNA would be best employed as a long-term repository (thousands or millions of years). How data-containing DNA might be packaged and how the data might be encrypted, with particular attention to the encryption of written information, is also discussed. Various encryption issues are touched on, such as how data-containing DNA might be differentiated from genetic material, error detection, data compression and reading frame location. Finally, this article broaches the difficulty of constructing very large pieces of DNA in the laboratory and highlights some complications that might arise when attempting to transmit DNA-encrypted data to recipients who are a long period of time in the future. PMID- 11412927 TI - Electrophoretic mobility anomalies associated with PCR amplification of the intergenic spacer region between 16S and 23S ribosomal RNA genes of Fusobacterium necrophorum. AB - PCR amplification of the intergenic spacer region (ISR) between 16S and 23S rRNA genes among subspecies of the anaerobic bacterium Fusobacterium necrophorum gave identical patterns, with two forms of ISR identified. However, extra bands resulting from anomalous electrophoretic mobility of amplified DNA fragments with certain primer combinations were encountered. Therefore, PCR assays relying solely on banding patterns may be unreliable, and supporting sequence analysis is essential for correct culture identification. PMID- 11412948 TI - Electronic control of helical chirality. AB - Chiral phenomena are common in living systems. Despite the fact that development of materials has often been inspired by chemistry from the biological world, materials that take advantage of inherent chirality have found relatively few applications. It is therefore probable that much remains to be gained from novel applications of molecular, macromolecular and supramolecular chirality. Among the most intriguing recent advances in studies of chiral materials is the development of mechanisms to control the shape and properties of chiral molecules. Photo induced helical chirality inversions have been studied for several years and significant achievements have been reported. Recently, electronically triggered systems have drawn significant attention. These technologies offer the potential for development of novel materials that take advantage of photonic or electronic modulation of molecular recognition, optical or mechanical properties. PMID- 11412949 TI - Delivering on the promise of bone morphogenetic proteins. AB - The advent of bone growth factors has been widely anticipated since their successful production using recombinant DNA technology. Bone morphogenetic proteins (BMPs) are an important class of bone growth factors and will be the focus of this article. In the near future these therapeutics might revolutionize how clinicians treat such diverse orthopedic applications as the healing of broken bones, increasing bone density lost through aging, and strengthening the spine. These potent proteins require application directly at the site of repair via a delivery system. The choice of delivery system has a profound effect on the clinical outcome. In the past decade, researchers have focused on developing efficient delivery systems and advancing these factors from the bench to the clinic. PMID- 11412950 TI - Recent developments in computational proteomics. AB - The mapping of the human genome was completed earlier this year and efforts are underway to understand the role of gene products (i.e. proteins) in biological pathways and human disease and to exploit their functional roles to derive protein therapeutics and protein-based drugs. A key component to the next revolution in the 'post-genomic' era will be the increasingly widespread use of protein structure in rational experimental design. Improvements in quality, availability and utility of large-scale 3D and 4D protein structural information are enabling a revolution in rational design, having particular impact on drug discovery and optimization. New computational methodologies now yield modeled structures that are, in many cases, quantitatively comparable with crystal structures, at a fraction of the cost. PMID- 11412951 TI - The genetic structures of a range of Douglas-fir provenance collections after planting in different European countries, assessed with two biochemical marker systems. AB - Genetic diversities of Douglas-fir provenance trials planted on several European sites were compared using both isozyme and terpene markers. A principal coordinate analysis based on similarity coefficients calculated from isozyme data indicated that, with the exception of two populations, differences between populations were small. There were no consistent trends in the variation in allele frequencies between populations. Most populations contained an excess of homozygotes, perhaps due to high selection pressure. Terpene composition was analysed in two resin systems, and showed a decreasing level of population diversity by planting site in the order: Spain-France-England-Scotland-Italy. Thus isozymes, which are generally considered to be neutral markers, indicated a modest degree of genetic drift due to sampling effects, while terpenes showed that some reduction in genetic diversity had occurred due to local selective pressures. PMID- 11412952 TI - Staphylococcus aureus MDR efflux pump inhibitors from a Berberis and a Mahonia (sensu strictu) species. AB - Bioactive fractionation, based on multi-drug resistance (MDR) pump inhibition in Staphylococcus aureus, resulted in the isolation of the active inhibitors 5' methoxyhydnocarpin-D from leaves of Berberis (formerly Mahonia) trifoliolata and pheophorbide a from Berberis fendleri. The hydnocarpin derivative was not found in the latter species. Pheophytin a (the phytol derivative of pheophorbide a) was identified from both species, but it proved to have no MDR pump inhibitory activity. The somewhat uncommon, and inactive, flavonoid tricin was identified from B. trifoliolata. The occurrence of a flavonolignan in Mahonia-tpe species and its absence in Berberis sensu strictu may provide a chemical differentiation between the two groups which are now recombined on the basis of DNA studies. The strong bacterial efflux pump inhibition of pheophorbide a could be of importance as a plant defense against natural pathogens. PMID- 11412953 TI - Allozyme characterisation of four Mediterranean evergreen oak species. AB - Polymorphism at 11 enzyme loci was used to compare the four evergreen oak species of the Mediterranean Basin and to establish their taxonomical relationships. As a comparison, two evergreen oak species from Tibet, located in the primary distribution area of evergreen oaks, were analysed at the same loci. Cladistic analyses led to the separation of a cluster of four species, namely the three Mediterranean Q. ilex, Q. coccifera and Q. alnifolia and the Tibetan Q. aquifolioides. The other Tibetan species, Q. semicarpifolia, and Q. suber, from the Western Mediterranean Basin, were very distinct genetically from the four other species. These results were not consistent with previous taxonomical treatment, based on morphology alone, which classified Q. ilex and Q. coccifera in "ilex group" (=subgenus Sclerophyllodrys O. Schwartz), and the other four species in "cerris group" (subgenus Cerris, according to Schwartz). Allozyme variation has thus provided useful information to clarify the taxonomy of evergreen oaks. PMID- 11412954 TI - Alkaloids of Erythroxylum (Erythroxylaceae) species from Southern Brazil. AB - A new alkaloid identified as 3beta,6beta-ditigloyloxynortropane as 3beta,7beta ditigloyloxynortropane, 4-hydroxyhygrinic acid, methylecgonidine and tropacocaine have been isolated from the leaves of Erythroxylum argentinum. The new structure was established by means of spectroscopic techniques. Four other species E. deciduum, E. microphyllum, E. pelleterianum and E. cuneifolium collected in the state of Rio Grande do Sul were screened for methylecgonidine and tropacocaine by CG/MS. Two of these plants contain tropacocaine and two contain methylecgonidine. GC/MS analysis of all 5 species for cocaine proved fruitless. The chemotaxonomic significance of these results is discussed. PMID- 11412955 TI - Chemosystematics of taxa from the Leontodon section Oporinia. AB - A chemosystematic study of the subgenus Oporinia of the genus Leontodon (Asteraceae) was performed, using flavonoids and phenolic acids in the flowerheads as diagnostic characters. A total of 44 samples from nine different Oporinia taxa were analyzed. Five luteolin-derivatives (luteolin, luteolin 7-O beta-D-gentiobioside, luteolin 7-O-beta-D-glucoside, luteolin 7-O-beta-D glucuronide, and luteolin 4'-O-beta-D-glucoside) and four caffeic acid derivatives (caffeoyl tartaric acid, chlorogenic acid, cichoric acid, and 3,5 dicaffeoylquinic acid) were identified in crude extracts by means of HPLC retention times, on-line UV spectra and on-line MS spectra. Quantification of these compounds was performed by HPLC, using quercetin as internal standard. The data obtained were processed by Principal Component Analysis, resulting in the formation of five different clusters. These clusters were taxonomically interpretable and are in good agreement with the morphologically based system of the genus Leontodon. PMID- 11412956 TI - Chemotaxonomy on the leaf constituents of Thujopsis dolabrata Sieb. et Zucc. Analysis of neutral extracts (diterpene hydrocarbon). AB - The leaf terpenes in Thujopsis dolabrata from all regions in Japan were analyzed using GC and GC-MS. The results show that the major constituents of the diterpene hydrocarbon fraction in this conifer are dolabradiene, hibaene, rimuene, 13-epi dolabradiene and abietatriene. There were wide variations in the contents of the constituents among individuals and habitats. The results also show that T. dolabrata trees from 34 habitats can be classified into three groups based on the composition of the leaf diterpene hydrocarbon. PMID- 11412957 TI - Flavonoids from Hyparrhenia hirta Stapf (Poaceae) growing in Tunisia. PMID- 11412958 TI - Cyclopropyl amino acids of Amanita. PMID- 11412960 TI - Tagetes tenuifolia Cav. (Asteraceae): some chemosystematic implications of their flavonoids. PMID- 11412959 TI - Secondary metabolites of Scilla plumbea, Ledebouria cooperi and Ledebouria ovatifolia (Hyacinthaceae). PMID- 11412961 TI - Comparative chemical composition of the resinous exudates from Senecio adenotrichius and S. viscosissimus. PMID- 11412962 TI - Sesquiterpene lactones from Lugoa revoluta. PMID- 11412963 TI - The identification of fusidic acid, a steroidal antibiotic from marine isolate of the fungus Stilbella aciculosa. PMID- 11412964 TI - Erythromycin biosynthesis. The 4-pro-S hydride of NADPH is utilized for ketoreduction by both module 5 and module 6 of the 6-deoxyerythronolide B synthase. AB - Incubation of chirally deuterated NADPH with 6-deoxyerythronolide B synthase (DEBS) modules 5 and module 6 and analysis of the derived triketide lactones established that the two ketoreductase domains, KR5 and KR6, are both specific for the 4-pro-S hydride of the nicotinamide cofactor. PMID- 11412965 TI - Synthesis and absolute configuration of Stellettadine A: a marine alkaloid that induces larval metamorphosis in ascidians. AB - Stellettadine A, a bisguanidinium alkaloid isolated from a marine sponge Stelletta sp. as an inducer of larval metamorphosis in ascidians, and its unnatural enantiomer were synthesized from the enantiomers of citronellal. The absolute configuration of stellettadine A was established as R. PMID- 11412966 TI - Active site of an aminoacyl-tRNA synthetase dissected by energy-transfer dependent fluorescence. AB - Aminoacyl-tRNA synthetases establish the rules of the genetic code by catalyzing attachment of amino acids to specific transfer RNAs (tRNAs) that bear the anticodon triplets of the code. Each of the 20 amino acids has its own distinct aminoacyl-tRNA synthetase. Here we use energy-transfer-dependent fluorescence from the nucleotide probe N-methylanthraniloyl dATP (mdATP) to investigate the active site of a specific aminoacyl-tRNA synthetase. Interaction of the enzyme with the cognate amino acid and formation of the aminoacyl adenylate intermediate were detected. In addition to providing a convenient tool to characterize enzymatic parameters, the probe allowed investigation of the role of conserved residues within the active site. Specifically, a residue that is critical for binding could be distinguished from one that is important for the transition state of adenylate formation. Amino acid binding and adenylate synthesis by two other aminoacyl-tRNA synthetases was also investigated with mdATP. Thus, a key step in the synthesis of aminoacyl-tRNA can in general be dissected with this probe. PMID- 11412967 TI - Aromatic inhibitors of dehydroquinate synthase: synthesis, evaluation and implications for gallic acid biosynthesis. AB - The role of the active site metal in determining binding to 3-dehydroquinate synthase has been examined. Protocatechuic acid, catechol, and derivatives of these aromatics were synthesized that shared the common element of an ortho dihydroxylated benzene ring. Inhibition constants were determined for each aromatic as well as the variation of this inhibition as a function of whether Co(+2) or Zn(+2) was the active site metal ion. PMID- 11412968 TI - Synthesis and applications of a deep, asymmetric cavitand on a solid support. AB - The synthesis of an optically active cavitand and its attachment to a polystyrene support are described. The cavitand is used for selective recognition based on size and shape of guest molecules in solution. PMID- 11412969 TI - Methodology for optimizing functional miniature proteins based on avian pancreatic polypeptide using phage display. AB - Synthetic genes for avian pancreatic polypeptide (aPP) and for the miniature DNA binding protein PPBR4 were cloned and expressed on the surface of M13 bacteriophage. We anticipate that these constructs will have utility optimizing the properties of miniature proteins based on aPP that result from our previously described protein grafting procedure. PMID- 11412970 TI - Regio-reactive resin: a platform for orthogonal loading using the polymer backbone and cross-linker. AB - A new support for solid-phase combinatorial organic synthesis has been developed, which we term a regio-reactive resin (R(3)-resin). The resin is based on a unique hydroxyl-functionalized cross-linker readily synthesized in two steps starting from 4-hydroxybenzaldehyde. The cross-linker's ease of synthesis and high purity enables the preparation of gel-type resins with regio-reactive orthogonal loading sites. The resin's swelling properties were investigated, and its potential utility was demonstrated via orthogonal reactivity of the pendant and cross linker sites. PMID- 11412971 TI - DNA alkylation by leinamycin can be triggered by cyanide and phosphines. AB - Previous work has shown that alkylation of DNA by the antitumor agent leinamycin (1) is potentiated by reaction of the antibiotic with thiols. Here, it is shown that other soft nucleophiles such as cyanide and phosphines can also trigger DNA alkylation by leinamycin. Overall, the results suggest that reactions of cyanide and phosphines with leinamycin produce the oxathiolanone intermediate (2), which is known to undergo rearrangement to the DNA-alkylating episulfonium ion 4. PMID- 11412972 TI - alpha-Keto heterocycle inhibitors of fatty acid amide hydrolase: carbonyl group modification and alpha-substitution. AB - Two sets of novel analogues of the recently disclosed alpha-keto heterocycle inhibitors of fatty acid amide hydrolase (FAAH), the enzyme responsible for regulation of endogenous oleamide and anandamide, were synthesized and evaluated in order to clarify a role of the electrophilic carbonyl group and structural features important for their activity. Both the electrophilic carbonyl and the degree of alpha-substitution markedly affect inhibitor potency. PMID- 11412973 TI - Bio-inspired synthetic receptor molecules towards mimicry of vancomycin. AB - A 512-member library of bio-inspired synthetic receptor molecules was prepared featuring a triazacyclophane scaffold. The purpose of this scaffold was to orient three (identical) peptide 'binding arms' in order to mimic an antibiotic binding cavity as is present in the vancomycin antibiotics. The library was screened with D-Ala-D-Ala and D-Ala-D-Lac containing ligands, which are present in the cell wall precursors of pathogenic bacteria. Screening and validation led to identification of a synthetic receptor capable of binding these ligands. PMID- 11412974 TI - Conversion of cyclic nonaketides to lovastatin and compactin by a lovC deficient mutant of Aspergillus terreus. AB - Investigation of the post-PKS biosynthetic steps to the cholesterol-lowering agent lovastatin (1) using an Aspergillus terreus strain with a disrupted lovC gene, which is essential for formation of 4a,5-dihydromonacolin L (3), shows that 7 and 3 are precursors to 1, and demonstrates that lovastatin diketide synthase (lovF protein) does not require lovC. PMID- 11412975 TI - Fluorescent pigments of the retinal pigment epithelium and age-related macular degeneration. AB - The major hydrophobic fluorophore of the retinal pigment epithelium (RPE) is A2E, a pyridinium bis-retinoid derived from all-trans-retinal and phosphatidyl ethanolamine. The accumulation of fluorophores such as A2E is implicated in the pathogenesis of age-related macular degeneration (AMD), a disease associated with the deterioration of central vision and a leading cause of blindness in the elderly. Recent chemical and biological studies have provided insight into the synthesis and biosynthesis of A2E, the spectroscopic properties of this pigment, and the role of A2E and RPE cell death. PMID- 11412976 TI - Rationale for the observed COX-2/COX-1 selectivity of celecoxib from Monte Carlo simulations. AB - Computational studies have yielded an analysis of the contributions to the free energy difference between the binding of celecoxib to COX-1 and to COX-2. The energetic and structural results point to the Ile to Val mutation at residue 523 as the key contributor to COX-2 selectivity; unfavorable steric contact between a sulfonamide oxygen and the delta methyl group of Ile523 destabilizes the complex with COX-1. The His to Arg change at residue 513 is less significant. PMID- 11412977 TI - Amidino benzimidazole inhibitors of bacterial two-component systems. AB - Amidino benzimidazoles have been identified as inhibitors of the bacterial KinA/Spo0F two-component system (TCS). Many of these inhibitors exhibit good in vitro antibacterial activity against a variety of susceptible and resistant Gram positive organisms. The moiety at the 2-position of the benzimidazole was extensively modified. In addition, the regioisomeric benzoxazoles, heterocyclic replacements for the benzimidazole, have been synthesized and their activity against the TCS evaluated. PMID- 11412978 TI - Cell permeability as a parameter for lead generation in the protein Tyrosine kinase inhibition field. AB - Based on the inverse relationship between polar surface area and cell permeability and capitalizing on the properties of pyrrolopyrimidines 1 as protein tyrosine kinase inhibitors, pyrrolopyridones 2 were designed and synthesized as potential leads for the development of novel inhibitors with improved cell permeability properties. PMID- 11412979 TI - Biaryl ether retrohydroxamates as potent, long-lived, orally bioavailable MMP inhibitors. AB - A novel series of biaryl ether reverse hydroxamate MMP inhibitors has been developed. These compounds are potent MMP-2 inhibitors with limited activity against MMP-1. Select members of this series exhibit excellent pharmacokinetic properties with long elimination half-lives (7 h) and high oral bioavailability (100%). PMID- 11412980 TI - Discovery and characterization of the potent, selective and orally bioavailable MMP inhibitor ABT-770. AB - Modification of the biphenyl portion of MMP inhibitor 2a gave analogue 2i which is greater than 1000-fold selective against MMP-2 versus MMP-1. The stereospecific synthesis of both enantiomers of 2i was achieved beginning with (S)- or (R)-benzyl glycidyl ether. The (S)-enantiomer, 11 (ABT-770), is orally bioavailable and efficacious in an in vivo model of tumor growth. PMID- 11412981 TI - N-Me-pAB-Glu-gamma-Glu-gamma-Tyr(3-NO(2)): an internally quenched fluorogenic gamma-glutamyl hydrolase substrate. AB - A gamma-glutamyl tripeptide containing an internally quenched fluorophore has been synthesized and shown to be a substrate for recombinant rat gamma-glutamyl hydrolase. HPLC, LC-MS, and fluorescence spectra support the conclusion that selective hydrolysis occurs at the penultimate peptide bond. Preliminary data indicate that hydrolysis of this substrate can be monitored continuously to yield steady-state kinetic data. PMID- 11412982 TI - Cyclooxygenase-inhibitory and antioxidant constituents of the aerial parts of Antirhea acutata. AB - Two new compounds, (6S)-hydroxy-29-nor-3,4-seco-cycloart-4(30),24-dien-3-oic acid (1) and 8-[1-(3,4-dihydroxyphenyl)-3-methoxy-3-oxopropyl]epicatechin (3), were isolated by bioassay-guided fractionation from the aerial parts of Antirhea acutata (DC.) Urb. (Rubiaceae). Compound 1 showed moderate inhibitory activities in cyclooxygenase-1 and -2 assays (IC(50) 43.7 and 4.7 microM, respectively), while compound 3 was active in 1,1-diphenyl-2-picrylhydrazyl free-radical and cytochrome c reduction antioxidant assays (IC(50) 29.1 and 16.3 microM, respectively). Additionally, one further new compound was isolated, (3S,24S)-25 trihydroxy-9,19-cycloartane-29-oic acid (2), but this was inactive in the bioassay systems used. Compound 1 is based on the unprecedented 29-nor-3,4-seco cycloartane skeleton. PMID- 11412983 TI - New probes of the agonist binding site of metabotropic glutamate receptors. AB - The (2S,4R)- and (2S,4S)-4-hydroxyglutamates activate cloned mGlu(1a), mGlu(2), and mGlu(8a) receptors with different potencies. Best results were obtained with the (2S,4S) isomer being almost as potent as glutamate on mGlu(1a)R and mGlu(8a)R. Data are interpreted on the basis of the binding site model and X-ray structure. PMID- 11412984 TI - Bioisosteric determinants for subtype selectivity of ligands for heteromeric GABA(A) receptors. AB - The potency and efficacy of a series of bioisosterically modified GABA analogues were determined electrophysiologically using heteromeric GABA(A) receptors expressed in Xenopus oocytes. These agonist parameters were shown to be strongly dependent on the receptor subunit combination. On the other hand, the antagonist potencies of the classical GABA(A) antagonists SR 95531 (7) and BMC (8) and also of 5g and the phosphinic acid bioisosteres of 5a, compounds 5f and 6, were essentially independent of the receptor subunit combinations. PMID- 11412985 TI - Phenylethanolamine N-methyltransferase kinetics: bovine versus recombinant human enzyme. AB - Epinephrine (Epi) acts as a neurotransmitter in the brain, but its function therein is not well understood. Phenylethanolamine N-methyltransferase (PNMT) catalyzes the final step in the biosynthesis of Epi and is thus a pharmacological target to investigate the function of Epi in the central nervous system. The kinetic differences between bovine adrenal PNMT and human brain PNMT for a number of substrates and inhibitors are examined and the results reported. PMID- 11412986 TI - An unhydrolyzable analogue of N-(4-hydroxyphenyl)retinamide. synthesis and preliminary biological studies. AB - The synthesis of a nonhydrolyzable, carbon-linked analogue (4-HBR) of the retinoid N-(4-hydroxyphenyl)retinamide (4-HPR) using Umpolung methods is described. Preliminary studies of biological activity show 4-HBR is similar to 4 HPR in its actions although a potentially relevant and desirable difference is its reduced suppression of plasma vitamin A levels. These results show that 4-HPR does not have to be hydrolyzed to retinoic acid to produce its chemotherapeutic effects. PMID- 11412987 TI - Synthesis of novel sialylmimetics as biological probes. AB - Glycomimetics are increasingly being recognised as powerful tools in the search for novel compounds that possess useful biological properties. This paper describes our preliminary efforts towards the development of novel mimetics of sialic acid thioglycosides. These sialylmimetics are readily prepared and have been shown, in some instances, to have biological properties similar to sialic acid thioglycosides. PMID- 11412988 TI - The cyclohexene ring as bioisostere of a furanose ring: synthesis and antiviral activity of cyclohexenyl nucleosides. AB - The application of the bioisosteric concept between a furanose ring and a cyclohexene ring in the nucleoside field has led to the discovery of new potent antiviral agents. PMID- 11412989 TI - The binding of arylguanidines at 5-HT(3) serotonin receptors: a structure affinity investigation. AB - The 5-HT(3) receptor binding affinities of nine pairs of aryl-substituted arylguanidines and arylbiguanides were examined and the results suggest the likelihood that both classes of agents utilize common receptor binding features. The effects of structural modification were also examined using CoMFA. 1-(3,4,5 Trichlorophenyl)guanidine (5-HT(3) K(i)=0.7 nM) was identified as a very high affinity arylguanidine. The structures of the high-affinity arylguanidines are inconsistent with current 5-HT(3) pharmacophore models. PMID- 11412990 TI - Synthesis and evaluation of GGPP geometric isomers: divergent substrate specificities of FTase and GGTase I. AB - A stereocontrolled synthetic route has been used to prepare two of the geometric isomers of all-trans-GGPP. Neither of these isomers is effective substrates for mammalian GGTase I, but 3 is a potent inhibitor of this enzyme (IC(50)=100 nM). Surprisingly, both compounds are effective substrates for mammalian FTase. PMID- 11412991 TI - Synthesis of derivatives of the novel cyclophilin-binding immunosuppressant sanglifehrin A with reduced numbers of polar functions. AB - The syntheses and the biological activities of 53-deoxo sanglifehrin A (2) and 61 deoxy octahydrosanglifehrin A (3) are described. Compound 2 shows intracellular cyclophilin (CyP)-binding and immunosuppressive activity in the mixed-lymphocyte reaction (MLR) similar to that of sanglifehrin A (1). Compound 3 is much less active in the MLR despite unchanged intracellular CyP-binding. This indicates that the 53-keto group is not necessary for immunosuppressive activity, while the 61 hydroxy group is required. PMID- 11412992 TI - Novel alkylpolyamine analogues that possess both antitrypanosomal and antimicrosporidial activity. AB - A novel series of alkyl- or aralkyl-substituted polyamine analogues was synthesized containing a 3-7-3 polyamine backbone. These analogues were evaluated in vitro, and in one case in vivo, for activity as antitrypanosomal agents, and for activity against opportunistic infection caused by Microsporidia. Compound 21 inhibits trypanosomal growth with an IC(50) as low as 31nM, while compound 24 shows promising activity in vitro against trypanosomes, and against Microsporidia in vitro and in vivo. PMID- 11412995 TI - Yeast histone deposition protein Asf1p requires Hir proteins and PCNA for heterochromatic silencing. AB - BACKGROUND: Position-dependent gene silencing in yeast involves many factors, including the four HIR genes and nucleosome assembly proteins Asf1p and chromatin assembly factor I (CAF-I, encoded by the CAC1-3 genes). Both cac Delta asfl Delta and cac Delta hir Delta double mutants display synergistic reductions in heterochromatic gene silencing. However, the relationship between the contributions of HIR genes and ASF1 to silencing has not previously been explored. RESULTS: Our biochemical and genetic studies of yeast Asf1p revealed links to Hir protein function. In vitro, an active histone deposition complex was formed from recombinant yeast Asf1p and histones H3 and H4 that lack a newly synthesized acetylation pattern. This Asf1p/H3/H4 complex generated micrococcal nuclease--resistant DNA in the absence of DNA replication and stimulated nucleosome assembly activity by recombinant yeast CAF-I during DNA synthesis. Also, Asf1p bound to the Hir1p and Hir2p proteins in vitro and in cell extracts. In vivo, the HIR1 and ASF1 genes contributed to silencing the heterochromatic HML locus via the same genetic pathway. Deletion of either HIR1 or ASF1 eliminated telomeric gene silencing in combination with pol30--8, encoding an altered form of the DNA polymerase processivity factor PCNA that prevents CAF-I from contributing to silencing. Conversely, other pol30 alleles prevented Asf1/Hir proteins from contributing to silencing. CONCLUSIONS: Yeast CAF-I and Asf1p cooperate to form nucleosomes in vitro. In vivo, Asf1p and Hir proteins physically interact and together promote heterochromatic gene silencing in a manner requiring PCNA. This Asf1/Hir silencing pathway functionally overlaps with CAF-I activity. PMID- 11412996 TI - CDC-42 regulates PAR protein localization and function to control cellular and embryonic polarity in C. elegans. AB - BACKGROUND: The polarization of the anterior-posterior axis (A-P) of the Caenorhabditis elegans zygote depends on the activity of the par genes and the presence of intact microfilaments. Functional links between the PAR proteins and the cytoskeleton, however, have not been fully explored. It has recently been shown that in mammalian cells, some PAR homologs form a complex with activated Cdc42, a Rho GTPase that is implicated in the control of actin organization and cellular polarity. A role for Cdc42 in the establishment of embryonic polarity in C. elegans has not been described. RESULTS: To investigate the function of Cdc42 in the control of cellular and embryonic polarity in C. elegans, we used RNA mediated interference (RNAi) to inhibit cdc-42 activity in the early embryo. Here, we demonstrate that RNAi of cdc-42 disrupts manifestations of polarity in the early embryo, that these phenotypes depend on par-2 and par-3 gene function, and that cdc-42 is required for the localization of the PAR proteins. CONCLUSIONS: Our genetic analysis of the regulatory relationships between cdc-42 and the par genes demonstrates that Cdc42 organizes embryonic polarity by controlling the localization and activity of the PAR proteins. Combined with the recent biochemical analysis of their mammalian homologs, these results simultaneously identify both a regulator of the PAR proteins, activated Cdc42, and effectors for Cdc42, the PAR complex. PMID- 11412997 TI - CDC-42 controls early cell polarity and spindle orientation in C. elegans. AB - BACKGROUND: Generation of asymmetry in the one-cell embryo of C. elegans establishes the anterior--posterior axis (A-P), and is necessary for the proper identity of early blastomeres. Conserved PAR proteins are asymmetrically distributed and are required for the generation of this early asymmetry. The small G protein Cdc42 is a key regulator of polarity in other systems, and recently it has been shown to interact with the mammalian homolog of PAR-6. The function of Cdc42 in C. elegans had not yet been investigated, however. RESULTS: Here, we show that C. elegans cdc-42 plays an essential role in the polarity of the one-cell embryo and the proper localization of PAR proteins. Inhibition of cdc-42 using RNA interference results in embryos with a phenotype that is nearly identical to par-3, par-6, and pkc-3 mutants, and asymmetric localization of these and other PAR proteins is lost. We further show that C. elegans CDC-42 physically interacts with PAR-6 in a yeast two-hybrid system, consistent with data on the interaction of human homologs. CONCLUSIONS: Our results show that CDC 42 acts in concert with the PAR proteins to control the polarity of the C. elegans embryo, and provide evidence that the interaction of CDC-42 and the PAR 3/PAR-6/PKC-3 complex has been evolutionarily conserved as a functional unit. PMID- 11412998 TI - Sexual conflict selects for male and female reproductive characters. AB - BACKGROUND: Strict genetic monogamy leads to sexual harmony because any trait that decreases the fitness of one sex also decreases the fitness of the other. Any deviation from monogamy increases the potential for sexual conflict. Conflict is further enhanced by sperm competition, and given the ubiquity of this phenomenon, sexual conflict is rife. In support of theory, experimentally enforced monogamy leads to the evolution of sexual benevolence. In contrast, with multiple mating, males evolve traits causing massive female fitness reductions when female evolution is restrained. Theory also predicts increased investment in spermatogenesis when sperm competition risk is high. While this supposition has correlational support, cause and effect has yet to be firmly established. RESULTS: By enforcing monogamy or polyandry in yellow-dung-fly lines, we have shown experimentally that males from polyandrous treatments evolved larger testes. Furthermore, females from this treatment evolved larger accessory sex glands. These glands produce a spermicidal secretion, so larger glands could increase female ability to influence paternity. Using molecular techniques, we have shown that, consistent with this idea, males' success as second mates is reduced in females from the polyandrous treatment. Nevertheless, males from polyandrous lines achieve higher paternity during sperm competition, and this finding further supports the testis evolution patterns. CONCLUSIONS: These results provide direct experimental support for macroevolutionary patterns of testis size evolution. Furthermore, we have shown that sperm competition selects for traits likely to be important in sexual conflicts over paternity, a result only previously demonstrated in Drosophila melanogaster. PMID- 11412999 TI - Multiple roles of mouse Numb in tuning developmental cell fates. AB - BACKGROUND: Notch signaling regulates multiple differentiation processes and cell fate decisions during both invertebrate and vertebrate development. Numb encodes an intracellular protein that was shown in Drosophila to antagonize Notch signaling at binary cell fate decisions of certain cell lineages. Although overexpression experiments suggested that Numb might also antagonize some Notch activity in vertebrates, the developmental processes in which Numb is involved remained elusive. RESULTS: We generated mice with a homozygous inactivation of Numb. These mice died before embryonic day E11.5, probably because of defects in angiogenic remodeling and placental dysfunction. Mutant embryos had an open anterior neural tube and impaired neuronal differentiation within the developing cranial central nervous system (CNS). In the developing spinal cord, the number of differentiated motoneurons was reduced. Within the peripheral nervous system (PNS), ganglia of cranial sensory neurons were formed. Trunk neural crest cells migrated and differentiated into sympathetic neurons. In contrast, a selective differentiation anomaly was observed in dorsal root ganglia, where neural crest- derived progenitor cells had migrated normally to form ganglionic structures, but failed to differentiate into sensory neurons. CONCLUSIONS: Mouse Numb is involved in multiple developmental processes and required for cell fate tuning in a variety of lineages. In the nervous system, Numb is required for the generation of a large subset of neuronal lineages. The restricted requirement of Numb during neural development in the mouse suggests that in some neuronal lineages, Notch signaling may be regulated independently of Numb. PMID- 11413000 TI - Zebrafish pea3 and erm are general targets of FGF8 signaling. AB - Phenotypic analysis of both zebrafish and mouse has shown that fibroblast growth factor 8 (FGF8) is required for many developmental decisions. To further our understanding of the FGF8 signaling process, we sought to identify new transcriptional targets of the pathway. Here, we propose that two zebrafish ETS genes, pea3 and erm, are general targets of FGF8 signaling, based upon the following observations: both genes are expressed around all early FGF8 signaling sources, both genes are downregulated in fgf8 mutant embryos in all tissues known to require fgf8 function, a pharmacological inhibitor of the FGF pathway completely abolishes expression of both genes, and ectopic expression of fgf8 is sufficient to induce both genes. The finding that pea3 and erm are common transcriptional targets of FGF8 signaling suggests that they are general mediators of FGF8 signaling during development. In addition, we observed that pea3 is often expressed close to an FGF8 source, and erm is expressed in a broader domain. To test whether this differential expression is established by FGF8, we have induced FGF8 ectopically and show that it is sufficient to recapitulate the endogenous nested expression pattern of pea3 and erm. PMID- 11413001 TI - Activation of the anaphase-promoting complex and degradation of cyclin B is not required for progression from Meiosis I to II in Xenopus oocytes. AB - Sister chromatid separation and cyclin degradation in mitosis depend on the association of the anaphase-promoting complex (APC) with the Fizzy protein (Cdc20), leading to the metaphase/anaphase transition and exit from mitosis [1- 3]. In Xenopus, after metaphase of the first meiotic division, only partial cyclin degradation occurs, and chromosome segregation during anaphase I proceeds without sister chromatid separation [4--7]. We investigated the role of xFizzy during meiosis using an antisense depletion approach. xFizzy accumulates to high levels in Meiosis I, and injection of antisense oligonucleotides to xFizzy blocks nearly all APC-mediated cyclin B degradation and Cdc2/cyclin B (MPF) inactivation between Meiosis I and II. However, even without APC activation, xFizzy-ablated oocytes progress to Meiosis II as shown by cyclin E synthesis, further accumulation of cyclin B, and evolution of the metaphase I spindle to a metaphase II spindle via a disc-shaped aggregate of microtubules known to follow anaphase I [8]. Inhibition of the MAPK pathway by U0126 in antisense-injected oocytes prevents cyclin B accumulation beyond the level that is present at metaphase I. Full synthesis and accumulation can be restored in the presence of U0126 by the expression of a constitutively active form of the MAPK target, p90(Rsk). Thus, p90(Rsk) is sufficient not only to partially inhibit APC activity [7], but also to stimulate cyclin B synthesis in Meiosis II. PMID- 11413002 TI - Establishment of human embryonic stem cell-transfected clones carrying a marker for undifferentiated cells. AB - Human embryonic stem (ES) cells are pluripotent cell lines that have been derived from the inner cell mass (ICM) of blastocyst stage embryos [1--3]. They are characterized by their ability to be propagated indefinitely in culture as undifferentiated cells with a normal karyotype and can be induced to differentiate in vitro into various cell types [1, 2, 4-- 6]. Thus, human ES cells promise to serve as an unlimited cell source for transplantation. However, these unique cell lines tend to spontaneously differentiate in culture and therefore are difficult to maintain. Furthermore, colonies may contain several cell types and may be composed of cells other than pluripotent cells [1, 2, 6]. In order to overcome these difficulties and establish lines of cells with an undifferentiated phenotype, we have introduced a reporter gene that is regulated by a promoter of an ES cell-enriched gene into the cells. For the introduction of DNA into human ES cells, we have established a specific transfection protocol that is different from the one used for murine ES cells. Human ES cells were transfected with enhanced green fluorescence protein (EGFP), under the control of murine Rex1 promoter. The transfected cells show high levels of GFP expression when in an undifferentiated state. As the cells differentiate, this expression is dramatically reduced in monolayer cultures as well as in the primitive endoderm of early stage (simple) embryoid bodies (EBs) and in mature EBs. The undifferentiated cells expressing GFP can be analyzed and sorted by using a Fluorescence Activated Cell Sorter (FACS). Thus, we have established lines of human ES cells in which only undifferentiated cells are fluorescent, and these cells can be followed and selected for in culture. We also propose that the pluripotent nature of the culture is made evident by the ability of the homogeneous cell population to form EBs. The ability to efficiently transfect human ES cells will provide the means to study and manipulate these cells for the purpose of basic and applied research. PMID- 11413003 TI - Ablation of the metal ion-induced endocytosis of the prion protein by disease associated mutation of the octarepeat region. AB - The neurodegenerative spongiform encephalopathies, or prion diseases, are characterized by the conversion of the normal cellular form of the prion protein PrP(C) to a pathogenic form, PrP(Sc) [1]. There are four copies of an octarepeat PHGG(G/S)WGQ that specifically bind Cu(2+) ions within the N-terminal half of PrP(C) [2--4]. This has led to proposals that prion diseases may, in part, be due to abrogation of the normal cellular role of PrP(C) in copper homeostasis [5]. Here, we show that murine PrP(C) is rapidly endocytosed upon exposure of neuronal cells to physiologically relevant concentrations of Cu(2+) or Zn(2+), but not Mn(2+). Deletion of the four octarepeats or mutation of the histidine residues (H68/76 dyad) in the central two repeats abolished endocytosis, indicating that the internalization of PrP(C) is governed by metal binding to the octarepeats. Furthermore, a mutant form of PrP that contains nine additional octarepeats and is associated with familial prion disease [6] failed to undergo Cu(2+)-mediated endocytosis. For the first time, these results provide evidence that metal ions can promote the endocytosis of a mammalian prion protein in neuronal cells and that neurodegeneration associated with some prion diseases may arise from the ablation of this function due to mutation of the octarepeat region. PMID- 11413004 TI - Prism adaptation to rightward optical deviation improves postural imbalance in left-hemiparetic patients. AB - Left-hemiparetic patients show predominant postural imbalance as compared to right-hemiparetic patients. The right hemisphere is crucial for generating internal maps used for perceptual and premotor processing of spatial information. Predominant postural imbalance with right-brain damage could thus result from a distortion of an internal postural map. Well-known manifestations of distorted internal maps due to right-hemisphere lesions, such as hemineglect, may show improvement following prism adaptation shifting the visual field to the right. We therefore investigated the effect of prism adaptation on postural imbalance in left-hemiparetic patients. Three groups of five patients were either adapted to prisms deviating the visual field to the right or left or exposed to neutral prisms while performing reaching movements of the right arm. Postural imbalance was reduced only following prism adaptation to the right. Thus, brief adaptation (i.e., 3 min) to rightward-shifting prisms can dramatically improve postural imbalance. This result shows that the effect of exposure to prisms that horizontally shift the visual field to the right in a reaching task generalizes to the postural system, and it suggests an interaction between horizontal and vertical reference frames. This also supports the theory that predominant postural imbalance in patients with right-brain damage may be partly related to a distortion of an internal postural map. PMID- 11413005 TI - Axoneme-specific beta-tubulin specialization: a conserved C-terminal motif specifies the central pair. AB - Axonemes are ancient organelles that mediate motility of cilia and flagella in animals, plants, and protists. The long evolutionary conservation of axoneme architecture, a cylinder of nine doublet microtubules surrounding a central pair of singlet microtubules, suggests all motile axonemes may share common assembly mechanisms. Consistent with this, alpha- and beta-tubulins utilized in motile axonemes fall among the most conserved tubulin sequences [1, 2], and the beta tubulins contain a sequence motif at the same position in the carboxyl terminus [3]. Axoneme doublet microtubules are initiated from the corresponding triplet microtubules of the basal body [4], but the large macromolecular "central apparatus" that includes the central pair microtubules and associated structures [5] is a specialization unique to motile axonemes. In Drosophila spermatogenesis, basal bodies and axonemes utilize the same alpha-tubulin but different beta tubulins [6--13]. beta 1 is utilized for the centriole/basal body, and beta 2 is utilized for the motile sperm tail axoneme. beta 2 contains the motile axoneme specific sequence motif, but beta 1 does not [3]. Here, we show that the "axoneme motif" specifies the central pair. beta 1 can provide partial function for axoneme assembly but cannot make the central microtubules [14]. Introducing the axoneme motif into the beta 1 carboxyl terminus, a two amino acid change, conferred upon beta 1 the ability to assemble 9 + 2 axonemes. This finding explains the conservation of the axoneme-specific sequence motif through 1.5 billion years of evolution. PMID- 11413007 TI - Can cloning save endangered species? AB - Efforts to clone some rare, endangered and even extinct species have created controversy amongst conservation bodies who think resources could be much better directed. Kenneth Lee reports. PMID- 11413008 TI - Race quickens for the first human clone. AB - The dazzling creation of Dolly, the cloned sheep, led many states to legislate against the possibility of using similar technology to create human clones. But for some, this prize is proving too tempting to ignore. Michael Gross reports. PMID- 11413009 TI - Ancient chemistry fuels new biology. AB - An enormous new greenhouse project in southern Britain aims to heighten awareness of the human relationship with plants and the growing potential of plant-derived compounds to find new uses, reports Nigel Williams PMID- 11413010 TI - The Sac phosphatase domain. PMID- 11413011 TI - Cooperativity in transcriptional control. PMID- 11413012 TI - Meiotic recombination: breaking the genome to save it. AB - Recombination ensures the correct segregation of chromosomes to gametes during meiosis. Recent studies point to a universal mechanism for initiating meiotic recombination: the formation of double-strand DNA breaks by Spo11p. PMID- 11413013 TI - Ran GTPase cycle: oOne mechanism -- two functions. AB - The Ras-related GTPase Ran functions in nucleocytoplasmic transport by regulating interactions of transport receptors with transport cargo. Three recent studies suggest that Ran uses exactly the same mechanism to regulate spindle assembly during mitosis. PMID- 11413014 TI - Cognitive neuroscience: who to play at poker. AB - Every neuroscientist knows that emotions are as much to do with the head as the heart, but as a number of new studies show, the heart - or rather the body - and the brain are by no means independent purveyors of feeling and emotion. PMID- 11413015 TI - Bacterial pathogenesis: the answer to virulence is in the pore. AB - A wide variety of Gram-negative bacterial pathogens use a 'type III' protein secretion system to deliver bacterial virulence factors into host cells. Recent results suggest that Gram-positive pathogens may employ similar methods to deliver virulence factors into host cells. PMID- 11413016 TI - Circadian rhythms: mop up the clock! AB - All circadian clock genes discovered in Drosophila have mammalian counterparts with extensive sequence homology. Similarities and differences have been identified between insect and mammalian oscillators. Recent studies have shed new light on two mammalian clock components: Mop3 and Per2. PMID- 11413017 TI - Cytokinin receptor: just another histidine kinase. AB - The cytokinin family of plant hormones is involved in diverse aspects of plant growth and development in vivo and in culture. Two groups have recently shown that a two-component histidine kinase functions as a cytokinin receptor specifically required for vascular development. PMID- 11413018 TI - Synaptic plasticity: rush hour traffic in the AMPA lanes. AB - Recent experiments indicate that modification of synaptic strength may involve rapid regulation of vesicular traffic on the postsynaptic side of the synapse. The specific vesicular trafficking route taken by postsynaptic receptors appears to depend on the stimulus. PMID- 11413019 TI - Recombination: homologous recombination branches out. AB - Homologous recombination can be divided into three key steps: strand exchange, branch migration and resolution. The identification of a protein complex that catalyses branch migration and Holliday junction resolution argues that the mechanism of homologous recombination is conserved from bacteria to man. PMID- 11413020 TI - Mammalian development: axes in the egg? AB - An enduring but erroneous belief is that the post-fertilisation period is irrelevant for axis development in mammals. Two recent studies further undermine this belief. Is information for axial developmental encoded in the egg cortex? PMID- 11413021 TI - Mitogens match cell numbers to local demand. AB - Although the control of cell proliferation has been studied intensively at the level of the single cell, less is known about how cell numbers are controlled in developing populations and organs. Often, proliferation provides a pool of cells for organ construction, but the rate of this proliferation must be coordinated with patterning to avoid imbalances in cell numbers. Recent research on the development of the Drosophila eye and the proliferation signals (mitogens) can act to coordinate cell numbers. PMID- 11413022 TI - All's well that ends well. AB - A recent Cold Spring Harbour meeting(*) reviewed the latest progress on telomeres (the specialized structures that form the ends of chromosomes) and telomerase (the enzyme primarily responsible for their replication). Among the many aspects of telomere biology covered were strong sessions elaborating telomere replication and length regulation, telomerase structure and function, end-binding and telomere-associated proteins, DNA-damage-response proteins and telomerase independent telomere maintenance. PMID- 11413039 TI - How is actin polymerization nucleated in vivo? AB - Actin polymerization in vivo is dependent on free barbed ends that act as nuclei. Free barbed ends can arise in vivo by nucleation from the Arp2/3 complex, uncapping of barbed ends on pre-existing filaments or severing of filaments by cofilin. There is evidence that each mechanism operates in cells. However, different cell types use different combinations of these processes to generate barbed ends during stimulated cell motility. Here, I describe recent attempts to define the relative contributions of these three mechanisms to actin nucleation in vivo. The rapid increase in the number of barbed ends during stimulation is not due to any single mechanism. Cooperation between capping proteins, cofilin and the Arp2/3 complex is necessary for the development of protrusive force at the leading edge of the cell: uncapping and cofilin severing contributing barbed ends, whereas activity of the Arp2/3 complex is necessary, but not sufficient, for lamellipod extension. These results highlight the need for new methods that enable the direct observation of actin nucleation and so define precisely the relative contributions of the three processes to stimulated cell motility. PMID- 11413040 TI - Not such a dismal science: the economics of protein synthesis, folding, degradation and antigen processing. AB - There is a pronounced tendency among cell biologists to focus on qualitative aspects of cell physiology. The remarkable accomplishments of evolution in creating cells can only be fully appreciated, however, by combining this qualitative analysis with a quantitative assessment of cellular constituents and processes. Here, I consider the overall protein economy of cells as it relates to recent advances in understanding protein folding, ubiquitin-targeted proteasome mediated degradation of proteins and the generation of peptide ligands for major histocompatibility complex (MHC) class I molecules. PMID- 11413041 TI - Lighting up the cell surface with evanescent wave microscopy. AB - Evanescent wave microscopy, also termed total internal reflection fluorescence microscopy (TIR-FM), has shed new light on important cellular processes taking place near the plasma membrane. For example, this technique can enable the direct observation of membrane fusion of synaptic vesicles and the movement of single molecules during signal transduction. There has been a recent surge in the popularity of this technique with the advent of green-fluorescent protein (GFP) as a fluorescent marker and new technical developments. These technical developments and some of the latest applications of TIR-FM are the subject of this review. PMID- 11413042 TI - Toll receptors in innate immunity. AB - Innate immunity is the first-line host defense of multicellular organisms that rapidly operates to limit infection upon exposure to infectious agents. In addition, the cells and molecules operating during this early stage of the immune response in vertebrates have a decisive impact on the shaping of the subsequent adaptive response. Genetic studies initially performed in the fruitfly Drosophila and later in mice have revealed the importance of proteins of the Toll family in the innate immune response. We present here our current understanding of the role of this evolutionary ancient family of proteins that are thought to function as cytokine receptors (Toll in Drosophila) or pattern-recognition receptors (TLRs in mammals) and activate similar, albeit non-identical, signal-transduction pathways in flies and mammals. PMID- 11413044 TI - Profile: Pamela Silver [interview by Mari N Jensen]. PMID- 11413045 TI - Antioxidant properties of macrophages toward low-density lipoprotein. AB - Oxidative modification of low-density lipoprotein (LDL) has been implicated in atherosclerosis. Intensive scientific efforts over the last two decades have focused on the elucidation of the mechanisms by which LDL is oxidized in vivo. A wealth of in vitro studies has demonstrated that the cell types present in atherosclerotic lesions, including monocyte/macrophages, quantitatively one of the most important cell types in plaque development, promote LDL oxidation. The mechanisms of cellular prooxidant activities have been extensively investigated. Fewer studies have addressed possible protective properties of the cells in LDL oxidation. This review summarizes recent observations of antioxidant, and potentially antiatherogenic, activities of macrophages toward LDL, including macrophage-mediated detoxification of lipid and protein hydroperoxides, metal sequestration and the generation of compounds with antioxidant properties. These activities could contribute to the net effect of macrophages on deleterious LDL oxidation and to the complex role of these cells in lesion development. PMID- 11413046 TI - The urokinase receptor associated protein (uPARAP/endo180): a novel internalization receptor connected to the plasminogen activation system. AB - The urokinase-mediated plasminogen activation system plays a central role in the extracellular proteolytic degradation reactions in cancer invasion. In this review article we discuss a number of recent findings identifying a new cellular receptor protein, uPARAP, that interacts with components of this proteolytic system. uPARAP is a high molecular weight type-1 membrane protein, belonging to the macrophage mannose receptor protein family. On the surface of certain cells, uPARAP forms a ternary complex with the pro-form of the urokinase-type plasminogen activator (uPA) and its primary receptor (uPAR). While the biological consequences of this reaction have not yet been verified experimentally, a likely event is ligand internalization because uPARAP is a constitutively recycling internalization receptor. uPARAP also binds at least one component, collagen type V, in the extracellular matrix meshwork, pointing to a potential role in proteolytic substrate presentation. Additional ligands have been proposed, including collagenase-3 and glycoproteins capable of interacting with one of the multiple carbohydrate recognition-type domains of uPARAP. In various adult tissues uPARAP is present on fibroblasts, macrophages and a subset of endothelial cells. In fetal tissues the protein has also been demonstrated in certain bone forming regions. Hypotheses on the physiological function of uPARAP include regulatory roles in extracellular proteolysis. This type of function would be likely to direct the local turnover of proteases and their substrate degradation products and thus may add to the complicated interplay between several cell types in governing restricted tissue degradation. PMID- 11413047 TI - Factor VIIIa-induced gene expression: potential implications in pathophysiology. AB - Binding of factor VIIa (VIIa) to tissue factor (TF) triggers the coagulation cascade. Several studies indicate a possible role for TF-VIIa in other cellular functions, in addition to coagulation, under pathophysiological conditions. Recent studies suggest that TF-VIIa mediates cell signaling by two distinct mechanisms, TF cytoplasmic domain-dependent and TF-VIIa protease activity dependent. In this review, we focus upon TF-VIIa protease-induced gene expression and how the altered gene expression could potentially affect various cellular processes. PMID- 11413048 TI - Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) in atherogenesis. AB - Lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) is a type-II membrane protein belonging to the C-type lectin family molecules, which can act as a cell surface endocytosis receptor for atherogenic oxidized LDL (Ox-LDL). LOX 1 is synthesized as a 40 kDa precursor protein with N-linked high mannose-type carbohydrate, which is further glycosylated and processed into a 50 kDa mature form. LOX-1 expression is not constitutive but can be induced by proinflammatory, oxidative, and mechanical stimuli. In addition to endothelial cells, macrophages and activated vascular smooth muscle cells express LOX-1. In vivo, endothelial cells covering early atherosclerotic lesions and macrophages and smooth muscle cells accumulated in the intima of advanced atherosclerotic plaques express LOX 1. LOX-1 is cleaved at membrane proximal extracellular domain by some protease activities and released from the cell surface. Measurement of soluble LOX-1 in vivo may provide novel diagnostic strategy for the evaluation and prediction of atherosclerosis and vascular diseases. PMID- 11413049 TI - Cubilin and megalin: partners in lipoprotein and vitamin metabolism. AB - Members of the low-density lipoprotein receptor (LDLR) family are unrivalled for their ability to endocytose and target ligands to lysosomes for degradation. Their endocytic and catabolic functions make them essential to homeostatic regulation of the level and activity of their ligands in biological fluids and interstitial spaces. Over the last few years it has become evident that the endocytic function of members of the LDLR family is employed by other kinds of cell surface receptors. Recently, the low-density lipoprotein receptor related protein-2 (megalin) was shown to act in concert with cubilin, a receptor for high density lipoproteins (HDL)/apolipoprotein A-I (apoA-I), intrinsic factor-vitamin B12 and albumin to mediate ligand endocytosis. In this article, we review the state of knowledge pertaining to cubilin and megalin, emphasizing their joint roles in both lipoprotein and vitamin metabolism. PMID- 11413050 TI - SPARC and the kidney glomerulus: matricellular proteins exhibit diverse functions under normal and pathological conditions. AB - In the last decade, numerous studies have emphasized the important functions that matricellular proteins subserve during angiogenesis, wound healing, and the maintenance of organ and tissue integrity. Matricellular proteins are defined as a group of secreted regulatory macromolecules that are not structural components of the extracellular matrix (ECM) but rather mediate interactions between the ECM and cells. One of these matricellular proteins, termed SPARC (secreted protein acidic and rich in cysteine), is produced during the process of wound healing and is prominent in several types of injury. An excessive deposition of glomerular matrix and an elevated proliferation of certain glomerular cells characterize a variety of kidney diseases. The proliferation of these cells is associated typically with the remodeling process that occurs after kidney injury, and is, at least in part, modulated by the altered expression of ECM, various growth factors, and the elevated production of matricellular proteins (e.g., SPARC). The secretion of one or more of the matricellular proteins can lead to expansion of the glomerular basement membrane, infiltration of immunocompetent cells, and, in some cases, to a reversal of the pathological condition. However, these proteins can also contribute collectively to renal fibrosis, glomerulosclerosis, glomerulonephritis, and the eventual loss of renal function. The purpose of this review is to evaluate the multiple functions of SPARC in the kidney glomerulus under normal and pathological conditions. PMID- 11413051 TI - Dual regulation of the cerebral microvasculature by epoxyeicosatrienoic acids. AB - Epoxyeicosatrienoic acids (EETs) are lipid metabolites that are synthesized in vascular endothelial cells. They are released by stimulation of their muscarinic receptors, and induce vaso-relaxation of cerebral blood vessels. In addition, cytochrome P450 epoxygenase enzymes, which catalyze the formation of epoxyeicosatrienoic acids, especially after stimulation by the excitatory neurotransmitter glutamate, are present in astrocytes, an abundant cell type in the brain that extends foot processes onto the cerebral microvessels. Using a modification of an efficient, recently developed, fluorescent assay, we have detected the presence of EETs in endothelial cells cultured from the cortex of rat brains as well as in neonatal astrocytes. We propose that both these cell types provide a dual supply of EETs to increase cerebral blood flow in order to meet systemic as well as localized nutrient demands of cells in the brain. PMID- 11413052 TI - Apoptosis of smooth muscle cells is not silent: Fas/FADD initiates a program of inflammatory gene expression. AB - Unlike necrosis, apoptosis is classically considered to be "silent," (i.e., self contained and non-inflammatory). In this review, we describe the system that we developed to regulate apoptosis of smooth muscle cells (SMC) in vivo. We have used this system to demonstrate that SMC apoptosis initiated by FADD or by Fas ligation includes a specific program of expression of pro-inflammatory genes. We discuss how this conclusion can be reconciled with reports that Fas plays an anti inflammatory role in vascular lesions. PMID- 11413053 TI - Introduction: mapping and ablation of rhythm abnormalities in pediatric patients. PMID- 11413054 TI - Atrioventricular node reentry tachycardia in pediatric patients. AB - Atrioventricular node reentry tachycardia (AVNRT) is a significant cause of paroxysmal supraventricular tachycardia (SVT) in the pediatric population. Symptoms can include palpitations, chest pain, fatigue, light-headedness and syncope. AVNRT is a reentry tachycardia that is comprised of dual conduction pathways through the AV node. On electrocardiogram, AVNRT usually manifests as a regular tachycardia with a narrow QRS complex and P waves that are either absent or distort the terminal portion of the QRS complex. Electrophysiology study will reveal dual AV node pathways: a fast pathway with a short AH interval and a long effective refractory period (ERP); and a slow pathway with a longer AH interval and a shorter ERP. During tachycardia, electrophysiologic signals will reveal conduction up the midline. Introduction of premature ventricular contractions and measurement of the HA interval during SVT can help distinguish AVNRT from a SVT utilizing an accessory pathway. Radiofrequency catheter ablation (RFA) has been used increasingly in children as treatment for AVNRT. The initial approach to RFA of AVNRT was modification of AV fast pathway conduction by lesions placed near the anterosuperior aspect of the triangle of Koch, known as the anterior approach method. However, this technique was associated with a significant risk of complete AV block. Now, the posterior approach slow pathway modification is used more commonly, which positions the ablation catheter along the tricuspid annulus immediately anterior to the coronary sinus ostium. This has been associated with a lower risk of complete AV block. Using this technique, RFA should be considered the method of choice for curative therapy of AVNRT in pediatric patients. PMID- 11413055 TI - Radiofrequency catheter ablation of left-sided accessory pathways in pediatric patients. AB - In many cases, radiofrequency catheter ablation has replaced the long-term use of antiarrhythmic medication for symptomatic tachycardia, and has all but eliminated arrhythmia surgery. The most common substrate for radiofrequency catheter ablation in pediatric patients is atrioventricular (AV) reentry tachycardia due to a concealed or manifest accessory pathway. Accessory pathways are distributed unevenly along the right and left atrioventricular valve annuli, and left-sided accessory pathways are most common. Although some centers advocate an abbreviated diagnostic and mapping approach to both concealed and manifest left-sided accessory pathways, most still use a complete electrophysiological evaluation and complex catheter manipulation for mapping, followed by the application of radiofrequency energy. Left-sided accessory pathways may be approached from the transatrial approach, the retrograde aortic approach, or less commonly from within the coronary sinus. Each approach has proven to be associated with success, but also with a distinct set of risks. Possibly because left-sided accessory pathways are most common, catheter ablation of this substrate has proven highly successful and has the lowest risk of recurrence. However, recent data also suggest that this substrate is associated with greater risk of complications than of right-sided accessory pathways or pathways located in the posteroseptal region. The following report reviews some of the recently described diagnostic and mapping techniques, success rates, risks and follow-up data in pediatric patients undergoing radiofrequency catheter ablation of left-sided accessory pathways. PMID- 11413056 TI - Radiofrequency ablation of right-sided accessory pathways in pediatric patients. AB - Right free-wall and septal accessory pathways encompass the full spectrum of accessory pathway electrophysiology and are situated in complex anatomical arrangements. Understanding this diversity of physiology is necessary for the successful and safe elimination of these connections with transcatheter radiofrequency ablation. When radiofrequency catheter ablation of these pathways is attempted in children, anatomical relationships often become more complex, and spatial constraints require more adaptive techniques than in adults. It is clear that considerable progress has been made with radiofrequency catheter ablation, such that it is now first-line therapy for most children who have been diagnosed with one of the broad spectrum of clinical manifestations that result from the presence of these accessory connections. This review will discuss how accessory pathway electrophysiology and anatomy impact the clinical syndromes observed in children, and how these factors, as well as others particular to children, determine the approach, results and potential long-term consequences of radiofrequency catheter ablation of right-sided accessory pathways in the pediatric population. PMID- 11413057 TI - Intra-atrial reentry tachycardia in pediatric patients. AB - Intra-atrial reentry tachycardia (IART), also known as atrial flutter, is a major problem in pediatric cardiology and adult congenital cardiology. Patients have significant morbidity and even mortality associated with this arrhythmia. The use of antiarrhythmic medications has been disappointing in this population. Ablation techniques are being developed which offer some advantages over the use of medication. These techniques include: sophisticated mapping using entrainment, electro-anatomic and non-contact methods for assessment of the anatomy and the reentrant circuit; radiofrequency ablation methods which allow for the creation of linear and transmural lesions; and new methods for assessment of the effects of ablation which focus on the documentation of the creation of a new line of block. These new techniques provide hope for more effective ablation procedures and the possibility of definitive cure of atrial flutter in many patients in this population. PMID- 11413058 TI - Mapping and ablation of ventricular tachycardia in children and adolescents. AB - Ventricular tachycardia is a rare arrhythmia in the pediatric population that occurs in structurally normal and abnormal hearts. The treatment of ventricular tachycardia is aimed at reducing symptoms and the risk of sudden death. The intracardiac electrophysiology study and radiofrequency ablation have come to the forefront of diagnostic and therapeutic options for patients with ventricular arrhythmias. The indications for radiofrequency ablation are debated, but it has been shown to be an effective and safe means of eliminating ventricular tachycardia. The purpose of this review is to outline the mechanisms and electrophysiologic characteristics of ventricular tachycardia, discuss the forms common to the pediatric population and summarize the indications, success and complications of radiofrequency ablation for ventricular tachycardia. PMID- 11413059 TI - Computational tools in rehabilitation of erectile dysfunction. AB - Erectile dysfunction (ED) is defined as the inability to achieve and maintain an erection adequate for satisfactory intercourse. It is a common problem among approximately 50% of men between the ages of 40 and 70. Erectile dysfunction is not only stressful to both the affected individual and his partner, but it can also negatively affect self-esteem. Biomechanical models have recently been developed to study both the structural and hemodynamic factors involved in normal and pathological erectile conditions. These computational models, which are reviewed in the present paper, allow for better understanding of the mechanisms acting in ED and provide a suitable basis for development of state-of-the-art interdisciplinary treatment approaches aimed to improve the quality of life for these men. PMID- 11413060 TI - Fuzzy clustering of gait patterns of patients after ankle arthrodesis based on kinematic parameters. AB - Kinematic parameters for 10 normal subjects and 10 patients with ankle arthrodesis are grouped using the fuzzy cluster paradigm. The features chosen for clustering are Euler angles of the sagittal plane in the hindfoot, the forefoot and combined hindfoot and forefoot joints. Gait patterns are identified using information provided by cluster validity techniques, giving three, three and two clusters for the hindfoot, forefoot and combined hindfoot and forefoot joints, respectively. The cluster centers represent distinct walking strategies adopted by normal subjects and patients after ankle arthrodesis. Utilizing angle values normalized by gait cycle, it is possible to classify any subject and to generate an individual's membership value for each of the clusters. The clinical utility of the fuzzy clustering approach is demonstrated with data for subjects with ankle arthrodesis, where changes in membership of the clusters provide an objective technique for measuring changes of gait pattern after ankle arthrodesis. This approach can be adopted to study other clinical entities where different cluster centers would be established using the algorithm provided in this study. PMID- 11413061 TI - Adaptive independent component analysis of multichannel electrogastrograms. AB - The electrogastrogram (EGG), a cutaneous measurement of gastric electrical activity, can be severely contaminated by endogenous biological noise sources such as respiratory signal. Therefore it is important to establish effective artifact removal methods. In this paper, a novel blind signal separation method with a flexible non-linearity is introduced and applied to extract the gastric slow wave from multichannel EGGs. Simulation results show that our algorithm is able to separate a wide range of source signals, including mixtures of Gaussian sources. On real data, we demonstrate the successful applications of our procedure to extract the gastric slow wave from multichannel EGGs. As a result, the extracted clean gastric slow wave can be used to facilitate further analysis, e.g. as a reference signal for multichannel adaptive enhancement of the EGG. PMID- 11413062 TI - Flow mixing and fluid residence times in a model of a ventricular assist device. AB - To provide a reliable tool for predicting the flow in a clinical sac-type ventricular assist device (VAD), a simplified model VAD was developed prior to this study for the purpose of experimental validation of computer simulations of the time-varying flow on a moving grid. At selected locations in the flow field, flow patterns, vortex core trajectories and velocities were compared between the numerical and physical models during one full pumping cycle. The computer simulations showed qualitatively very good agreement with the experiment. Quantitatively, the agreement was less favourable. In order to investigate flow mixing and to evaluate fluid residence times in the model VAD a new tool has been developed: the numerical simulations were extended by injecting a scalar into the flow domain. These simulations allow investigation of the flow mixing qualitatively by visualisation and quantification of fluid residence times. Experimental assessment of the numerical results using dye injection proved to be favourable. The numerical results have been extended to include some prediction of pressure which have been compared with experimental measurements. PMID- 11413063 TI - Investigation of the nonlinearity in the heart rate dynamics. AB - This paper presents the application of a neural network to predict human heart rate. Electrocardiograms were measured from 5 healthy adult human subjects and 5 data sets were constructed calculating instantaneous heart rate from the measured signal. The nonlinear radial basis function neural network was applied to have a one step ahead prediction of the 1000 point heart rate. The results of the prediction are compared to that obtained by a linear autoregressive model. The results show that the neural network performs better than the autoregressive model in predicting heart rate for 2 data sets while for the other 3 data sets the performance of the two models is statistically similar. This indicates that the heart rate may be controlled nonlinearly by the autonomic nervous system. PMID- 11413064 TI - A new hybrid algorithm for ECG signal compression based on the wavelet transformation of the linearly predicted error. AB - This paper describes a hybrid technique based on the combination of wavelet transform and linear prediction to achieve very effective electrocardiogram (ECG) data compression. First, the ECG signal is wavelet transformed using four different discrete wavelet transforms (Daubechies, Coiflet, Biorthogonal and Symmlet). All the wavelet transforms are based on dyadic scales and decompose the ECG signals into five detailed levels and one approximation. Then, the wavelet coefficients are linearly predicted, where the error corresponding to the difference between these coefficients and the predicted ones is minimized in order to get the best predictor. In particular, the residuals of the wavelet coefficients are uncorrelated and hence can be represented with fewer bits compared to the original signal. To further increase the compression rate, the residual sequence obtained after linear prediction is coded using a newly developed coding technique. As a result, a compression ratio (Cr) of 20 to 1 is achieved with percentage root-mean square difference (PRD) less than 4%. The algorithm is compared to an alternative compression algorithm based on the direct use of wavelet transforms. Experiments on selected records from the MIT-BIH arrhythmia database reveal that the proposed method is significantly more efficient in compression. The proposed compression scheme may find applications in digital Holter recording, in ECG signal archiving and in ECG data transmission through communication channels. PMID- 11413065 TI - Compression of ECG signals by optimized quantization of discrete cosine transform coefficients. AB - This paper presents an ECG compressor based on optimized quantization of Discrete Cosine Transform (DCT) coefficients. The ECG to be compressed is partitioned in blocks of fixed size, and each DCT block is quantized using a quantization vector and a threshold vector that are specifically defined for each signal. These vectors are defined, via Lagrange multipliers, so that the estimated entropy is minimized for a given distortion in the reconstructed signal. The optimization method presented in this paper is an adaptation for ECG of a technique previously used for image compression. In the last step of the compressor here proposed, the quantized coefficients are coded by an arithmetic coder. The Percent Root-Mean Square Difference (PRD) was adopted as a measure of the distortion introduced by the compressor. To assess the performance of the proposed compressor, 2-minute sections of all 96 records of the MIT-BIH Arrhythmia Database were compressed at different PRD values, and the corresponding compression ratios were computed. We also present traces of test signals before and after the compression/decompression process. The results show that the proposed method achieves good compression ratios (CR) with excellent reconstruction quality. An average CR of 9.3:1 is achieved for PRD equal to 2.5%. Experiments with ECG records used in other results from the literature revealed that the proposed method compares favorably with various classical and state-of-the-art ECG compressors. PMID- 11413066 TI - Slippage of a spinous process hook during flexion in a flexible fixation system for the lumbar spine. AB - This study was undertaken to measure the amount of slippage of a spinous process hook (that forms part of a flexible fixation system) during flexion. Human cadaveric lumbar spines (10) were fitted with the device. A rig was designed to apply flexural displacements to a spine using a materials testing machine. Spherical markers were attached to the spine and hook. As a spine was flexed a digital video camera was used to record the positions of the markers. The movements of the markers were measured using interactive computer software to assess any slippage of the spinous process hook. During flexion the overall mean hook slippage was measured to be 0.10mm (standard deviation 0.04mm). The mean hook slippage, for each of the 10 specimens, was in the range of 0.05-0.14mm. The results imply that slippage of a spinous process hook during flexion is small. PMID- 11413067 TI - A force platform for large human displacements. AB - This paper provides details of a force platform assembly suitable for measuring the magnitude, position and direction of ground reaction forces for manual handling tasks involving several footsteps in any horizontal direction. An approach to increase the natural frequency with the use of a vibration absorber is used; this frequency was found to be 41+/-2 Hz. The results show a maximum error of 22 N (2% of the full scale output) when a person performs repetitive movements from a squat position to a full extension and back to the squat position. Static tests show the maximal errors to be, vertically, 1.5% of the full scale output (1818+/-33 N), and horizontally, 4.9% of the full scale output (1177+/-6 N). PMID- 11413068 TI - Heart failure 2001 meeting, Barcelona, Spain, 9-12 June 2001. Abstracts. PMID- 11413070 TI - Partnerships for the health of the public. PMID- 11413071 TI - Looks aren't everything. FFR B4 U PTCA. PMID- 11413072 TI - Nitroglycerin and delayed preconditioning in humans: yet another new mechanism for an old drug? PMID- 11413073 TI - Cell cycle and cell migration: new pieces to the puzzle. PMID- 11413074 TI - Rapid three-dimensional echocardiography : clinically feasible alternative for precise and accurate measurement of left ventricular volumes. AB - BACKGROUND: Clinical applicability of conventional ultrasonographic systems using mechanical adapters for 3D echocardiographic imaging has been limited by long acquisition and processing times. We developed a rapid (6-s) acquisition technique that collects apical tomograms using a continuously internally rotating transthoracic transducer. This study was performed to examine the clinical feasibility of rapid-acquisition 3D echocardiography to estimate left ventricular end-diastolic and end-systolic volumes using electron-beam computed tomography as the reference standard. Methods and Results-We collected a series of 6 to 11 apical echocardiographic tomograms, depending on heart rate, in 11 patients. There was good correlation, low variability, and low bias between rapid 3D echocardiography and electron-beam computed tomography for measuring left ventricular end-diastolic volume (r=0.96; standard error of the estimate, 21.34 mL; bias, -4.93 mL) and left ventricular end-systolic volume (r=0.96; standard error of the estimate, 14.78 mL; bias, -6.97 mL). CONCLUSIONS: The rapid acquisition 3D echocardiography extends the use of a multiplane, internally rotating handheld transducer so that it becomes a precise and clinically feasible tool for assessing left ventricular volumes and function. A rapid-image acquisition time of 6 s would allow repeated image collection during the course of a clinical echocardiographic examination. Additional work must address rapid and automated data processing. PMID- 11413075 TI - Increase in circulating endothelial progenitor cells by statin therapy in patients with stable coronary artery disease. AB - BACKGROUND: Therapeutic neovascularization may constitute an important strategy to salvage tissue from critical ischemia. Circulating bone marrow-derived endothelial progenitor cells (EPCs) were shown to augment the neovascularization of ischemic tissue. In addition to lipid-lowering activity, hydroxymethyl glutaryl coenzyme A reductase inhibitors (statins) reportedly promote the neovascularization of ischemic tissue in normocholesterolemic animals. Methods and Results-Fifteen patients with angiographically documented stable coronary artery disease (CAD) were prospectively treated with 40 mg of atorvastatin per day for 4 weeks. Before and weekly after the initiation of statin therapy, EPCs were isolated from peripheral blood and counted. In addition, the number of hematopoietic precursor cells positive for CD34, CD133, and CD34/kinase insert domain receptor was analyzed. Statin treatment of patients with stable CAD was associated with an approximately 1.5-fold increase in the number of circulating EPCs by 1 week after initiation of treatment; this was followed by sustained increased levels to approximately 3-fold throughout the 4-week study period. Moreover, the number of CD34/kinase insert domain receptor-positive hematopoietic progenitor cells was significantly augmented after 4 weeks of therapy. Atorvastatin treatment increased the further functional activity of EPCs, as assessed by their migratory capacity. CONCLUSION: The results of the present study define a novel mechanism of action of statin treatment in patients with stable CAD: the augmentation of circulating EPCs with enhanced functional activity. Given the well-established role of EPCs of participating in repair after ischemic injury, stimulation of EPCs by statins may contribute to the clinical benefit of statin therapy in patients with CAD. PMID- 11413076 TI - Benefit of glycoprotein IIb/IIIa inhibition in patients with acute coronary syndromes and troponin t-positive status: the paragon-B troponin T substudy. AB - BACKGROUND: Troponin T (TnT) is valuable for short- and long-term risk stratification of patients with acute coronary syndromes (ACS). It also may predict which ACS patients will benefit from glycoprotein (GP) IIb/IIIa blockade. METHODS AND RESULTS: We prospectively studied 1160 patients with non-ST-segment elevation ACS randomized in PARAGON-B to receive lamifiban, an intravenous GP IIb/IIIa antagonist, or placebo. TnT levels were obtained before study treatment began and 24 to 72 hours later; assays were performed by a blinded core laboratory. At baseline, 40.2% of patients were TnT-positive (>/=0.1 ng/mL); these patients were older and more often male or smokers. Patients positive at baseline had a significantly higher rate of the primary end point (composite of death, myocardial [re]infarction, or severe recurrent ischemia at 30 days; odds ratio, 1.5; 95% CI, 1.1 to 2.1) than those who were TnT-negative. Lamifiban was associated with significant reduction in the primary end point (from 19.4% to 11.0%, P=0.01) among TnT-positive patients but not among TnT-negative patients (11.2% for placebo versus 10.8% for lamifiban, P=0.86; P=0.08 for test of interaction between TnT status and treatment assignment). This pattern held for the end points of death alone and death or myocardial (re)infarction at 30 days. Peak TnT level at 48 hours did not differ with lamifiban treatment. CONCLUSIONS: TnT predicts poor short-term outcomes in non-ST-segment elevation ACS. Treatment benefit with lamifiban is limited almost exclusively to TnT-positive patients, reducing 30-day adverse outcomes to a rate nearly identical to that of negative patients. PMID- 11413077 TI - Speed of intracranial clot lysis with intravenous tissue plasminogen activator therapy: sonographic classification and short-term improvement. AB - BACKGROUND: Arterial recanalization precedes clinical improvement or may lead to hemorrhage or reperfusion injury. Speed of clot lysis was not previously measured in human stroke. METHODS AND RESULTS: Transcranial Doppler (TCD) and the National Institutes of Health Stroke Scale (NIHSS) were used to monitor consecutive patients receiving intravenous tissue plasminogen activator (tPA), before tPA bolus and at 24 hours. Patients with complete or partial recanalization of the middle cerebral or basilar artery on TCD were studied. Recanalization was classified a priori as sudden (abrupt appearance of a normal or stenotic low resistance signal), stepwise (flow improvement over 1 to 29 minutes), or slow (>/=30 minutes). Recanalization was documented in 43 tPA-treated patients (age 68+/-17 years; NIHSS score 16.8+/-6, median 15 points). tPA bolus was given at a mean of 135+/-61 minutes after symptom onset. Recanalization began at a median of 17 minutes and was completed at 35 minutes after tPA bolus, with mean duration of recanalization of 23+/-16 minutes. Recanalization was sudden in 5, stepwise in 23, and slow in 15 patients. Faster recanalization predicted better short-term improvement (P=0.03). At 24 hours, 80%, 30%, and 13% of patients in these respective recanalization groups had NIHSS scores of 0 to 3. Symptomatic hemorrhage occurred in only 1 patient, who had stepwise recanalization 5.5 hours after stroke onset. Slow or partial recanalization with dampened flow signal was found in 53% of patients with total NIHSS scores >10 points at 24 hours (P=0.01). Complete recanalization (n=25) occurred faster (median 10 minutes) than partial recanalization (n=18; median 30 minutes; P=0.0001). CONCLUSIONS: Rapid arterial recanalization is associated with better short-term improvement, mostly likely because of faster and more complete clot breakup with low resistance of the distal circulatory bed. Slow (>/=30 minutes) flow improvement and dampened flow signal are less favorable prognostic signs. These findings may be evaluated to assist with selection of patients for additional pharmacological or interventional treatment. PMID- 11413078 TI - Transdermal estrogen replacement therapy decreases sympathetic activity in postmenopausal women. AB - BACKGROUND: Menopause heralds a dramatic increase in incident hypertension, suggesting a protective effect of estrogen on blood pressure (BP). In female rats, estrogen has been shown to decrease sympathetic nerve discharge (SND) and BP. SND, however, has not been recorded during estrogen replacement therapy (ERT) in humans. Methods and Results-In 12 normotensive postmenopausal women, we conducted a randomized crossover placebo-controlled study to test whether chronic ERT caused a sustained decrease in SND and BP. Twenty-four-hour ambulatory BP, SND, and arterial baroreflex sensitivity were measured before and after 8 weeks of transdermal estradiol (200 microgram/d), oral conjugated estrogens (0.625 mg/d), or placebo. To test the acute effects of estrogen on SND, additional studies were performed in the same women receiving intravenous conjugated estrogens or sublingual estradiol. After 8 weeks of transdermal ERT, the basal rate of SND decreased by 30% (from 40+/-4 to 27+/-4 bursts per minute, P=0.0001) and ambulatory diastolic BP fell by 5+/-2 mm Hg (P=0.0003). In contrast, SND and BP were unaffected either by 8 weeks of oral ERT or by acute estrogen administration. Neither transdermal nor oral ERT had any effects on baroreflex sensitivity. CONCLUSIONS: In normotensive postmenopausal women, chronic transdermal ERT decreases SND without augmenting arterial baroreflexes and causes a small but statistically significant decrease in ambulatory BP. Sympathetic inhibition is evident only with chronic rather than acute estrogen administration, implying a genomic mechanism of action. Because the effects of transdermal ERT are larger than those of oral ERT, the route of administration may be an important consideration in optimizing the beneficial effects of ERT on BP and overall cardiovascular health. PMID- 11413079 TI - Estrogen replacement therapy improves baroreflex regulation of vascular sympathetic outflow in postmenopausal women. AB - BACKGROUND: Menopausal estrogen loss has been associated with increased cardiovascular disease in postmenopausal women. However, the link between estrogen and cardiovascular disease remains unclear. Some data suggest estrogen mediates its effect through changes in arterial pressure and its regulation. However, the data available in older women are equivocal regarding estrogen's ability to reduce resting arterial pressure or to improve its regulation. METHODS AND RESULTS: We studied 11 healthy, postmenopausal women before and after 6 months of estrogen administration. Arterial pressure was measured by brachial auscultation and finger photoplethysmography. Vascular sympathetic nerve activity was measured in the peroneal nerve by microneurography, and the slope of the relations between changes in heart period, sympathetic activity, and arterial pressure caused by bolus infusions of nitroprusside and phenylephrine were used as an index of baroreflex gain. Estrogen therapy did not change systolic pressure (128+/-2 versus 123+/-2 mm Hg) or cardiac-vagal baroreflex gain (6.6+/-0.9 versus 6.7+/-0.7 ms/mm Hg). However, vascular sympathetic baroreflex gain was increased (-4.6+/-0.6 versus -7.4+/-1.0 arbitrary integrated units/mm Hg; P=0.02). CONCLUSION: These findings suggest long-term estrogen replacement therapy has effects on cardiovascular regulation that may not be reflected in resting arterial pressures. PMID- 11413080 TI - Cytomegalovirus infection with interleukin-6 response predicts cardiac mortality in patients with coronary artery disease. AB - BACKGROUND: Prospective data relating previous exposure to cytomegalovirus (CMV) to the risk of cardiac mortality are controversial. We investigated the effect of previous exposure to CMV infection on the risk of future cardiac disease-related death in relation to an underlying inflammatory response. METHODS AND RESULTS: coronary angiography was performed in 1134 subjects, and 989 patients with documented coronary artery disease were studied prospectively. CMV-IgG titers and interleukin (IL)-6 levels were measured before angiography. Increasing titers of CMV correlated with the elevation of IL-6 levels (P<0.001) after adjustment for possible confounders. All patients were followed up for a median of 3.1 years (maximum 4.3 years). During follow-up, 96 patients died, 70 of cardiac disease. Overall, CMV seropositivity was not related to cardiac mortality after adjustment for confounding variables (P=0.19). In contrast, in patients with elevated IL-6 levels (>/=11.9 pg/mL, median level), CMV seropositivity was independently associated with a 3.2-fold (95% CI 1.4 to 7.3, P=0.007) increase in risk of future cardiac death, whereas in individuals without IL-6 elevation, previous CMV infection had no effect on cardiac mortality. CONCLUSIONS: MV seropositivity in patients with an inflammatory response is independently associated with future cardiac mortality, whereas this association is lost in patients who do not demonstrate an inflammatory response. These data support the hypothesis that the atherosclerotic effects of CMV are mediated through an underlying inflammatory response. PMID- 11413081 TI - Oxygenated carotenoid lutein and progression of early atherosclerosis: the Los Angeles atherosclerosis study. AB - BACKGROUND: Carotenoids are hypothesized to explain some of the protective effects of fruit and vegetable intake on risk of cardiovascular disease. The present study assessed the protective effects of the oxygenated carotenoid lutein against early atherosclerosis. METHODS AND RESULTS: EPIDEMIOLOGY: Progression of intima-media thickness (IMT) of the common carotid arteries over 18 months was determined ultrasonographically and was related to plasma lutein among a randomly sampled cohort of utility employees age 40 to 60 years (n=480). Coculture: The impact of lutein on monocyte response to artery wall cell modification of LDL was assessed in vitro by quantification of monocyte migration in a coculture model of human intima. Mouse models: The impact of lutein supplementation on atherosclerotic lesion formation was assessed in vivo by assigning apoE-null mice to chow or chow plus lutein (0.2% by weight) and LDL receptor-null mice to Western diet or Western diet plus lutein. IMT progression declined with increasing quintile of plasma lutein (P for trend=0.007, age-adjusted; P=0.0007, multivariate). Covariate-adjusted IMT progression (mean+/-SEM) was 0.021+/-0.005 mm in the lowest quintile of plasma lutein, whereas progression was blocked in the highest quintile (0.004+/-0.005 mm; P=0.01). In the coculture, pretreatment of cells with lutein inhibited LDL-induced migration in a dose-dependent manner (P<0.05). Finally, in the mouse models, lutein supplementation reduced lesion size 44% in apoE-null mice (P=0.009) and 43% in LDL receptor-null mice (P=0.02). CONCLUSIONS: These epidemiological, in vitro, and mouse model findings support the hypothesis that increased dietary intake of lutein is protective against the development of early atherosclerosis. PMID- 11413082 TI - Fractional flow reserve to determine the appropriateness of angioplasty in moderate coronary stenosis: a randomized trial. AB - BACKGROUND: PTCA of a coronary stenosis without documented ischemia at noninvasive stress testing is often performed, but its benefit is unproven. Coronary pressure-derived fractional flow reserve (FFR) is an invasive index of stenosis severity that is a reliable substitute for noninvasive stress testing. A value of 0.75 identifies stenoses with hemodynamic significance. METHODS AND RESULTS: In 325 patients for whom PTCA was planned and who did not have documented ischemia, FFR of the stenosis was measured. If FFR was >0.75, patients were randomly assigned to deferral (deferral group; n=91) or performance (performance group; n=90) of PTCA. If FFR was <0.75, PTCA was performed as planned (reference group; n=144). Clinical follow-up was obtained at 1, 3, 6, 12, and 24 months. Event-free survival was similar between the deferral and performance groups (92% versus 89% at 12 months and 89% versus 83% at 24 months) but was significantly lower in the reference group (80% at 12 months and 78% at 24 months). In addition, the percentage of patients free from angina was similar between the deferral and performance groups (49% versus 50% at 12 months and 70% versus 51% at 24 months) but was significantly higher in the reference group (67% at 12 and 80% at 24 months). CONCLUSIONS: In patients with a coronary stenosis without evidence of ischemia, coronary pressure-derived FFR identifies those who will benefit from PTCA. PMID- 11413083 TI - Delayed preconditioning-mimetic action of nitroglycerin in patients undergoing coronary angioplasty. AB - BACKGROUND: Experimental studies suggest that the cardioprotective effects of the late phase of ischemic preconditioning (PC) can be mimicked pharmacologically. However, to date, no drug has been tested with respect to its ability to elicit a late PC effect in humans. As a consequence, clinical exploitation of the powerful anti-stunning and anti-infarct actions of late PC has been elusive thus far. METHODS AND RESULTS: A total of 66 patients were randomized to receive a 4-hour intravenous infusion of nitroglycerin (NTG) or normal saline; on the following day, they underwent percutaneous transluminal coronary angioplasty (three 2 minute balloon inflations 5 minutes apart). Measurements of ST-segment shifts (intracoronary and surface ECGs), regional wall motion (quantitative 2D echocardiography), and chest pain score indicated that the infusion of NTG 24 hours before angioplasty rendered the myocardium relatively resistant to ischemia and that the degree of this cardioprotective effect was comparable to that afforded by the ischemia associated with the first balloon inflation in control subjects (early phase of ischemic PC). Collateral flow (estimated from a pressure derived index) did not differ between control and NTG-pretreated patients, indicating that the enhanced tolerance to ischemia in NTG-pretreated patients cannot be accounted for by baseline differences in collateral function. CONCLUSIONS: NTG protects human myocardium against ischemia 24 hours after its administration. To the best of our knowledge, this is the first report that a late PC effect can be recruited pharmacologically in humans. The results suggest that prophylactic administration of nitrates could be a novel approach to the protection of the ischemic myocardium in patients. PMID- 11413084 TI - Atrioventricular node modification in patients with chronic atrial fibrillation: role of morphology of RR interval variation. AB - BACKGROUND: This study evaluates the role of RR interval distribution pattern as an outcome predictor of radiofrequency (RF) modification of atrioventricular (AV) node in chronic atrial fibrillation (AF) and attempts to elucidate the likely mechanism of rate control. METHODS AND RESULTS: Sixty-five patients with chronic AF underwent AV node modification. The RR interval distribution pattern was derived from 24-hour ECG recordings obtained before and after the procedure. The preablation pattern was bimodal (B) in 36 patients (55%) and unimodal (U) in 29 patients (45%). After the modification procedure, the B pattern shifted to U (78%) or became modified B (22%). The mean number of RF pulses delivered and the fluoroscopy time were n=8+/-5 and 24+/-11 minutes, respectively, in patients with B pattern versus n=18+/-7 and 45+/-17 minutes in patients with U pattern (P<0.001 for both). The location of successful ablation was posteroseptal and lower midseptal in 26 patients (81%) with B pattern versus 2 (13%) with U pattern (P<0.001). Mean and maximal ventricular rates and heart rate at peak exercise were reduced after the procedure in both groups (P<0.001 for all). Long-term success rate, AV block incidence, and pacemaker implantation rate were 89%, 0%, and 8%, respectively, in patients with B pattern versus 52% (P<0.001), 21% (P=0.006), and 48% (P<0.001) in patients with U pattern. CONCLUSIONS: RF modification of the AV node is expected to be more effective, safe, and expeditious in patients with chronic AF and B RR interval distribution pattern. Posterior atrionodal input ablation may be the prevailing mechanism of rate control in these patients, whereas U-pattern patients may benefit from partial injury to the AV node. PMID- 11413085 TI - Differential effects of nifedipine and co-amilozide on the progression of early carotid wall changes. AB - BACKGROUND: Common carotid artery intima-media thickness (IMT) progression was compared between 4 years of treatment with nifedipine and diuretic. METHODS AND RESULTS: This study, ancillary to the International Nifedipine GITS Study: Intervention as a Goal in Hypertension Treatment (INSIGHT), involved nifedipine 30 mg or co-amilozide (hydrochlorothiazide 25 mg and amiloride 2.5 mg) with optional subsequent titration. Among 439 randomized hypertensive patients, 324 had >/=1 year of follow-up (intent-to-treat group), and 242 completed follow-up (until-end-of-study group). Ultrasonography was performed at baseline, 4 months later, and then every year. Central computerized reading provided far-wall IMT, diameter, and cross-sectional area IMT (CSA-IMT). The primary outcome was IMT progression rate (slope of IMT-time regression). Secondary outcomes were changes from baseline (Delta) in IMT, diameter, and CSA-IMT. In the until-end-of-study population, between-treatment differences existed in IMT progression rate (P=0.002), Delta IMT (P=0.001), and Delta CSA-IMT (P=0.006), because IMT progressed on co-amilozide but not on nifedipine. In the intent-to-treat population, treatment differences existed in Delta IMT (P=0.004) and Delta CSA IMT (P=0.04) but not in IMT progression rate (P=0.09). Patients with >/=2, 3, or 4 years of follow-up showed treatment differences in IMT progression rate (P=0.04, 0.004, 0.007, respectively), Delta IMT (P=0.005, 0.001, 0.005), and Delta CSA-IMT (P=0.025, 0.013, 0.015). Diameter decreased more on co-amilozide than on nifedipine in the intent-to-treat population (P<0.05), whereas blood pressure decreased similarly on both treatments. CONCLUSIONS: A difference in early carotid wall changes is shown between 2 equally effective antihypertensive treatments. PMID- 11413086 TI - Functional blockade of platelet-derived growth factor receptor-beta but not of receptor-alpha prevents vascular smooth muscle cell accumulation in fibrous cap lesions in apolipoprotein E-deficient mice. AB - BACKGROUND: The vascular smooth muscle cell (VSMC) is the central cell component involved in the fibroproliferative response in atherogenesis. As the lesion advances, VSMCs migrate from the media into the subendothelial space, thereby forming fibrous plaque lesions. Platelet-derived growth factor (PDGF) has been known to be a potent chemoattractant and mitogen for SMCs, but the pathophysiological role of the 2 PDGF receptors, receptor-alpha (PDGFR-alpha) and receptor-beta (PDGFR-beta) in atherogenesis is poorly understood. To clarify this problem, we prepared antagonistic rat monoclonal antibodies, APA5 and APB5, against murine PDGFR-alpha and PDGFR-beta, respectively. METHODS AND RESULTS: Apolipoprotein E-deficient mice were fed a high-fat diet containing 0.3% cholesterol from 6 weeks of age and subjected to injection with 1 mg/d IP of either antibody from 12 to 18 weeks every other day. In the mice injected with APB5, the aortic atherosclerotic lesion size and the number of intimal VSMCs were reduced by 67% and 80%, respectively, compared with the control mice injected with irrelevant rat IgG. In contrast, the mice that received APA5 showed only minimal reduction of lesion size, and a large number of VSMCs were observed in the intima. In the intima of advanced lesions, APB5 immunolabeled VSMCs, whereas APA5 could detect VSMCs mainly in the media. CONCLUSIONS: These results indicate that PDGFR-beta plays a significant role in formation of fibrous atherosclerotic lesions and that regulation of the signal transduction through PDGFR-beta could affect atherogenesis in mice. PMID- 11413087 TI - Responses of GLUT4-deficient hearts to ischemia underscore the importance of glycolysis. AB - BACKGROUND: The ischemic heart is dependent on glycolysis for ATP generation, and therapies that increase glucose utilization during ischemia improve survival. Myocardial ischemia results in the translocation of the glucose transporter proteins GLUT1 and GLUT4 to the sarcolemma. The increased glucose entry via these transporters contributes to enhanced glycolysis during ischemia. METHODS AND RESULTS: To determine the role of GLUT4 in mediating increased glycolytic flux during ischemia, hearts from mice with cardiac-selective GLUT4 deficiency (G4H-/ ) were subjected to global low-flow ischemia. During normal perfusion, hearts from fed G4H-/- mice showed increased GLUT1-mediated glucose uptake, higher concentrations of glycogen and phosphocreatine, but delayed recovery after ischemia. When these compensatory changes were eliminated by a 20-hour fast, G4H /- hearts exhibited depressed glucose utilization during ischemia and developed profound and irreversible systolic and diastolic dysfunction associated with accelerated ATP depletion during ischemia and diminished regeneration of high energy phosphate compounds on reperfusion. CONCLUSIONS: GLUT4 is an important mediator of enhanced glycolysis during ischemia and represents an important protective mechanism against ischemic injury. PMID- 11413088 TI - Role for p27(Kip1) in Vascular Smooth Muscle Cell Migration. AB - BACKGROUND: Rapamycin is a potent inhibitor of smooth muscle cell (SMC) proliferation and migration. Rapamycin-mediated inhibition of SMC proliferation is associated with upregulation of the cyclin-dependent kinase inhibitor p27(Kip1). Previously, we showed that mixed embryonic fibroblasts obtained from p27(Kip1)(-/-) mice were relatively rapamycin-resistant, suggesting that p27(Kip1) plays an integral role in modulating the antiproliferative effects of rapamycin. We hypothesized that the antimigratory effect of rapamycin may also be mediated by p27(Kip1). METHODS AND RESULTS: Rapamycin (1 to 10 nmol/L) inhibited basic fibroblast growth factor-induced migration of wild-type (WT) but not p27(Kip1)(-/-) SMCs in a dose-dependent manner (P<0.05) in a modified Boyden chamber. The effects of rapamycin on aortic SMC explant migration were also studied with WT, p27(+/-), and p27(-/-) mice. Rapamycin 4 mg. kg(-1). d(-1) IP for 5 days inhibited SMC migration by 90% in the WT and p27(Kip1)(+/-) (P<0.05) but not p27(Kip1)(-/-) animals. CONCLUSIONS: Lack of p27(Kip1) reduces rapamycin mediated inhibition of SMC migration. These novel findings suggest a role for p27(Kip1) in the signaling pathway(s) that regulates SMC migration. PMID- 11413089 TI - Ablation of serotonin 5-HT(2B) receptors in mice leads to abnormal cardiac structure and function. AB - BACKGROUND: Identification of factors regulating myocardial structure and function is important to understand the pathogenesis of heart disease. Because little is known about the molecular mechanism of cardiac functions triggered by serotonin, the link between downstream signaling circuitry of its receptors and the heart physiology is of widespread interest. None of the serotonin receptor (5 HT(1A), 5-HT(1B), or 5-HT(2C)) disruptions in mice have resulted in cardiovascular defects. In this study, we examined 5-HT(2B) receptor-mutant mice to assess the putative role of serotonin in heart structure and function. METHODS AND RESULTS: We have generated G(q)-coupled 5-HT(2B) receptor-null mice by homologous recombination. Surviving 5-HT(2B) receptor-mutant mice exhibit cardiomyopathy with a loss of ventricular mass due to a reduction in number and size of cardiomyocytes. This phenotype is intrinsic to cardiac myocytes. 5-HT(2B) receptor-mutant ventricles exhibit dilation and abnormal organization of contractile elements, including Z-stripe enlargement and N-cadherin downregulation. Echocardiography and ECG both confirm the presence of left ventricular dilatation and decreased systolic function in the adult 5-HT(2B) receptor-mutant mice. CONCLUSIONS: Mutation of 5-HT(2B) receptor leads to a cardiomyopathy without hypertrophy and a disruption of intercalated disks. 5 HT(2B) receptor is required for cytoskeleton assembly to membrane structures by its regulation of N-cadherin expression. These results constitute, for the first time, strong genetic evidence that serotonin, via the 5-HT(2B) receptor, regulates cardiac structure and function. PMID- 11413090 TI - Effects of balloon injury on neointimal hyperplasia in streptozotocin-induced diabetes and in hyperinsulinemic nondiabetic pancreatic islet-transplanted rats. AB - BACKGROUND: The mechanisms of increased neointimal hyperplasia after coronary interventions in diabetic patients are still unknown. METHODS AND RESULTS: Glucose and insulin effects on in vitro vascular smooth muscle cell (VSMC) proliferation and migration were assessed. The effect of balloon injury on neointimal hyperplasia was studied in streptozotocin-induced diabetic rats with or without adjunct insulin therapy. To study the effect of balloon injury in nondiabetic rats with hyperinsulinemia, pancreatic islets were transplanted under the kidney capsule in normal rats. Glucose did not increase VSMC proliferation and migration in vitro. In contrast, insulin induced a significant increase in VSMC proliferation and migration in cell cultures. Furthermore, in VSMC culture, insulin increased MAPK activation. A reduction in neointimal hyperplasia was consistently documented after vascular injury in hyperglycemic streptozotocin induced diabetic rats. Insulin therapy significantly increased neointimal hyperplasia in these rats. This effect of hyperinsulinemia was totally abolished by transfection on the arterial wall of the N17H-ras-negative mutant gene. Finally, after experimental balloon angioplasty in hyperinsulinemic nondiabetic islet-transplanted rats, a significant increase in neointimal hyperplasia was observed. CONCLUSIONS: In rats with streptozotocin-induced diabetes, balloon injury was not associated with an increase in neointimal formation. Exogenous insulin administration in diabetic rats and islet transplantation in nondiabetic rats increased both blood insulin levels and neointimal hyperplasia after balloon injury. Hyperinsulinemia through activation of the ras/MAPK pathway, rather than hyperglycemia per se, seems to be of crucial importance in determining the exaggerated neointimal hyperplasia after balloon angioplasty in diabetic animals. PMID- 11413091 TI - Fibrinolysis for acute myocardial infarction: current status and new horizons for pharmacological reperfusion, part 2. PMID- 11413092 TI - Three-dimensional visualization of pulmonary thromboemboli in chronic thromboembolic pulmonary hypertension with multiple detector-row spiral computed tomography. PMID- 11413093 TI - Guide to anticoagulant therapy: Heparin : a statement for healthcare professionals from the American Heart Association. PMID- 11413094 TI - ACC/AHA guidelines for percutaneous coronary intervention (revision of the 1993 PTCA guidelines)-executive summary: a report of the American College of Cardiology/American Heart Association task force on practice guidelines (Committee to revise the 1993 guidelines for percutaneous transluminal coronary angioplasty) endorsed by the Society for Cardiac Angiography and Interventions. PMID- 11413095 TI - Primary oxalosis with cardiac involvement: echocardiographic features of an unusual form of cardiomyopathy. PMID- 11413096 TI - Bush Administration and the new senate converge on health care. PMID- 11413097 TI - Using skeletal muscle cells to repair ailing hearts. PMID- 11413099 TI - Prospects for intervention in gastric carcinogenesis: reversibility of gastric atrophy and intestinal metaplasia. PMID- 11413100 TI - GRP and Pavlov's dogs. PMID- 11413101 TI - Surviving too long in Crohn's disease. PMID- 11413102 TI - The subtleties of intestinal metaplasia. PMID- 11413103 TI - Methotrexate: a useful alternative in Crohn's disease? PMID- 11413104 TI - Conservation of the cag pathogenicity island is associated with vacA alleles and gastroduodenal disease in South African Helicobacter pylori isolates. AB - BACKGROUND: The development of clinically significant disease in South Africa is associated with the vacuolating cytotoxin gene (vacA) s1 genotype but not with the presence of the cytotoxin associated gene cagA. cagA occurs in >95% of South African isolates and is a variable marker for the entire cag pathogenicity island (PAI). AIM: To characterise the cagPAI in South African isolates and to investigate if structural variants of this multigene locus were associated with variations in vacA status and clinical outcome. PATIENTS AND METHODS: We studied 109 Helicobacter pylori strains (36 from patients with peptic ulceration, 26 with gastric adenocarcinoma, and 47 with no pathology other than gastritis) for differences in selected genes of the cagPAI and alleles of vacA by polymerase chain reaction. RESULTS: All strains were cagA(+). Sixty five (60%) strains had an intact contiguous cagPAI; 78% of peptic ulcer isolates, 73% of gastric adenocarcinoma isolates, but only 40% of gastritis alone isolates (p< 0.01). The entire cagII region was undetectable in 23% of gastritis alone isolates but in only 8% of peptic ulceration isolates (p<0.05). The vacA signal sequence and mid region demonstrated a strong relationship between the virulence associated vacA s1 (p<0.005) and vacA m1 (p=0.05) alleles and an intact cagPAI. CONCLUSION: Although a complete cagPAI was a feature of most infected individuals, deletions in the 5' region of this genetic locus were associated with gastritis alone and with the non-cytotoxic s2/m2 vacA genotype. PMID- 11413105 TI - Helicobacter pylori induced transactivation of SRE and AP-1 through the ERK signalling pathway in gastric cancer cells. AB - BACKGROUND AND AIMS: Helicobacter pylori infection induces expression of proinflammatory cytokines such as interleukin (IL)-8 and tumour necrosis factor alpha (TNF-alpha) in gastric mucosa, and their genes have AP-1 binding sites in the promoter region. c-Fos is important for transactivation of AP-1 which has SRE in the promoter region. We conducted this study to confirm H pylori induced transactivation of these binding sites. METHODS: Transactivation of SRE and AP-1 was evaluated in human gastric cancer cells TMK1 and MKN45 by luciferase reporter assay in transient transfection. We compared the effects of coculture with four H pylori strains, a cag pathogenicity island (PAI) positive strain TN2, its isogenic vacA negative (TN2-DeltavacA) or cagE negative (TN2-DeltacagE) mutants, and a cag PAI negative clinical isolate T68. Phosphorylation of ERK1/2, JNK, and c-Jun was measured by immunoblot, induction of IL-8 secretion by ELISA, and the effects of MEK by inhibitor U0126. RESULTS: Both SRE and AP-1 were transactivated by coculture with TN2. Although TN2-DeltavacA induced comparable transactivation, TN2-DeltacagE and T68 showed decreased transactivation of SRE (65% and 51%) and AP-1 (71% and 54%, respectively, of TN2). Heat killed TN2 or indirect contact using a permeable membrane inhibited transactivation. Levels of phosphorylated ERK1/2, JNK, and c-Jun were increased by coculture with TN2. MEK inhibitor U0126 reduced TN2 induced transactivation of SRE and AP1, as well as secretion of IL-8, by 83%, 87%, and 53%, respectively, of TN2. CONCLUSIONS: Transactivation of SRE and AP-1, through ERK/MAPK and JNK/SAPK cascades, respectively, was found in gastric cancer cells cocultured with H pylori. Direct contact with viable bacteria possessing intact cag PAI is a prerequisite for the onset of intracellular signalling leading to AP-1 transactivation. PMID- 11413106 TI - Regulation of gastric function by endogenous gastrin releasing peptide in humans: studies with a specific gastrin releasing peptide receptor antagonist. AB - BACKGROUND AND AIMS: The main goal of our study was to characterise the activity of BIM26226 as a peripheral gastrin releasing peptide (GRP) receptor antagonist in healthy human subjects and to determine if endogenous GRP is a physiological regulator of gastric acid secretion and gastrin release. METHODS: Our study consisted of three parts. In part I, subjects received saline or BIM26226 followed by graded doses of intravenous human GRP in a four period crossover design. In part II, subjects received BIM26226 or saline during oral meal ingestion or modified sham feeding. In part III, subjects received an acidified meal in the presence and absence of BIM26226 in a two period crossover design. In addition, gastrin and somatostatin mRNA were measured in biopsy specimens during saline and BIM26226 infusion. RESULTS: BIM26226 dose dependently inhibited GRP induced acid output. Acid secretion after oral liquid meal intake and sham feeding was significantly inhibited by BIM26226 (p<0.01) whereas plasma gastrin release remained unchanged. Gastrin and somatostatin mRNAs were not significantly different after saline or BIM26226. CONCLUSIONS: BIM26226 is a potent GRP antagonist in humans. Endogenous GRP may be a physiological regulator of gastric acid secretion. Gastrin release does not seem to be under the control of GRP. PMID- 11413107 TI - Age and alarm symptoms do not predict endoscopic findings among patients with dyspepsia: a multicentre database study. AB - INTRODUCTION: Symptoms of dyspepsia are common but most patients do not have major upper gastrointestinal pathology. Endoscopy is recommended for dyspeptic patients over the age of 45, or those with certain "alarm" symptoms. We have evaluated the effectiveness of age and "alarm" symptoms for predicting major endoscopic findings in six practising endoscopy centres. METHODS: Clinical variables of consecutive patients with dyspepsia symptoms undergoing upper endoscopy examinations were recorded using a common endoscopy database. Patients who had no previous upper endoscopy or barium radiography were included. Stepwise multivariate logistic regression was used to identify predictors of endoscopic findings. The accuracy of these for predicting endoscopic findings was evaluated with receiver operating characteristic analysis. The sensitivity and specificity of age thresholds from 30 to 70 years were evaluated. RESULTS: Major pathology (tumour, ulcer, or stricture) was found at endoscopy in 787/3815 (21%) patients with dyspepsia. Age, male sex, bleeding, and anaemia were found to be significant but weak independent predictors of endoscopic findings. A multivariate prediction rule based on these factors had poor predictive accuracy (c statistic=0.62). Using a simplified prediction rule of age > or =45 years or the presence of any "alarm" symptom, sensitivity was 87% and specificity was 26%. Increasing or decreasing the age cut off did not significantly improve the predictive accuracy. CONCLUSIONS: Age and the presence of "alarm" symptoms are not effective predictors of endoscopic findings among patients with dyspepsia. Better clinical prediction strategies are needed to identify patients with significant upper gastrointestinal pathology. PMID- 11413108 TI - Decreased Bax expression by mucosal T cells favours resistance to apoptosis in Crohn's disease. AB - BACKGROUND: Activated T cells are more susceptible to apoptosis than resting T cells. As intestinal T cells normally exhibit a higher state of activation, increased apoptosis may be necessary to maintain immune homeostasis in the specialised microenvironment of the mucosa. On the other hand, in Crohn's disease (CD) mucosal T cells are resistant to apoptosis, suggesting abnormal regulation of cell death mechanisms. AIMS: To investigate differences in expression of anti- and proapoptotic Bcl-2 family proteins, key regulators of apoptosis, between circulating and mucosal T cells, and possible alterations in CD. PATIENTS AND METHODS: Lamina propria T cells (LPT) were isolated from 10 control, seven CD, and eight ulcerative colitis (UC) patients, and peripheral blood T cells (PBT) from healthy volunteers. Purified T cells were stained intracellularly for Bcl-2, Bcl-x(L), and Bax, and mean fluorescence intensity measured by flow cytometry. RESULTS: Compared with PBT, the expression level of Bcl-2 and Bax, but not Bcl x(L), was significantly greater in LPT, resulting in lower Bcl-x(L)/Bax ratios. In PBT, Bax expression was highly and significantly correlated with both Bcl-2 and Bcl-x(L), but correlation with Bcl-2 was absent in LPT. Bax expression in CD, but not UC, LPT was significantly lower than in control LPT, resulting in a significantly higher Bcl-x(L)/Bax ratio. The significant correlation of Bcl-x(L) to Bax was preserved in CD, but not UC, LPT. CONCLUSIONS: Regulation of Bcl-2 family protein expression differs between circulating and mucosal T cells, probably underlying diverse survival potentials. In CD LPT, a low Bax expression and a high Bcl-x(L)/Bax ratio favour resistance to apoptosis and may contribute to the chronicity of inflammation. PMID- 11413109 TI - Interleukin 10 (Tenovil) in the prevention of postoperative recurrence of Crohn's disease. AB - BACKGROUND AND AIMS: New lesions of Crohn's disease occur early after ileal or ileocolonic resection and ileocolonic anastomosis. We performed a double blind controlled trial to evaluate the safety and tolerance of recombinant human interleukin 10 (IL-10; Tenovil) in subjects operated on for Crohn's disease. We also assessed the effect of Tenovil in preventing endoscopic recurrence 12 weeks after surgery. METHODS: Patients with Crohn's disease who underwent curative ileal or ileocolonic resection and primary anastomosis were randomised within two weeks after surgery to receive subcutaneous Tenovil 4 microg/kg once daily (QD) (n=22) or 8 microg/kg twice weekly (TIW) (n=21), or placebo (QD or TIW) (n=22). An ileocolonoscopy was performed after 12 weeks of treatment. RESULTS: Compliance was excellent. The most frequently observed adverse events were mild and moderate in severity and equally distributed across treatment groups. Thirty seven patients in the pooled Tenovil group and 21 patients in the pooled placebo group were evaluable by endoscopy. At 12 weeks, 11 of 21 patients (52%) in the placebo group had recurrent lesions compared with 17 of 37 patients (46%) in the Tenovil group (ns). The incidence of severe endoscopic recurrence was similar in both groups (9%). CONCLUSION: Tenovil treatment for 12 consecutive weeks in patients with Crohn's disease after intestinal resection was safe and well tolerated. No evidence of prevention of endoscopic recurrence of Crohn's disease by Tenovil was observed. PMID- 11413110 TI - Escherichia coli strains colonising the gastrointestinal tract protect germfree mice against Salmonella typhimurium infection. AB - BACKGROUND: Escherichia coli is part of the normal gastrointestinal microflora which exerts a barrier effect against enteropathogens. Several E coli strains develop a protective effect against other Enterobacteriaceae. AIMS: Two E coli strains, EM0, a human faecal strain, and JM105 K-12 were tested for their ability to prevent in vivo and in vitro infection by Salmonella typhimurium C5. METHODS: Inhibition of C5 cell invasion by E coli was investigated in vitro using Caco 2/TC7 cells. The protective effect of E coli was examined in vivo in germfree or conventional C3H/He/Oujco mice orally infected by the lethal strain C5. RESULTS: EMO expresses haemolysin and cytotoxic necrotising factor in vitro. In vitro, the two strains did not prevent the growth of C5 by secreted microcins or modified cell invasion of C5. In vivo, establishment of EM0 or JM105 in the gut of germfree mice resulted in a significant increase in the number of surviving mice: 11/12 and 9/12, respectively, at 58 days after infection (2x10(6)/mouse) versus 0/12 in control germfree group at 13 days after infection. Colonisation level and translocation rate of C5 were significantly reduced during the three days after infection. In contrast, no reduction in faecal C5 excretion was observed in C5 infected conventional mice (1x10(8)/mouse) receiving the EM0 or JM105 cultures daily. CONCLUSIONS: Establishment of E coli strains, which do not display antimicrobial activity, protects germfree mice against infection and delays the establishment of C5 in the gut. Possible mechanisms of defence are discussed. PMID- 11413111 TI - Involvement of nerves and calcium channels in the intestinal response to Clostridium difficile toxin A: an experimental study in rats in vivo. AB - BACKGROUND: The involvement of nerves and calcium channels in the intestinal response to Clostridium difficile toxin A (luminal concentration 1 or 15 microg/ml) was studied in the small intestine of rats in vivo. METHODS: Inflammation was quantified by estimating myeloperoxidase (MPO) activity in the intestinal lumen, extravascular accumulation of Evan's blue (EB) in the intestine, and number of red blood cells (RBCs) in veins in histological sections. Intestinal damage was estimated using a histological grading system. In some experiments net fluid transport was recorded using a gravimetric technique. RESULTS: In acutely denervated intestines, toxin A caused marked destruction of the villi, increased luminal release of MPO activity, and augmentation of intestinal content of EB and venous RBCs. Denervating the intestine 3-4 weeks prior to the actual experiment prevented the development of villus damage and significantly decreased the number of RBCs in intestinal veins in experiments with a low toxin concentration, whereas no effect was demonstrated on luminal MPO activity. Using a high toxin concentration, chronic denervation decreased only the number of RBCs. Pretreatment with hexamethonium (low toxin concentration; acute denervation) attenuated the effect of toxin A on morphology, luminal MPO activity, and number of RBCs. Pretreatment with nifedipine (low toxin concentration; acute denervation) significantly decreased intestinal MPO activity and number of RBCs. Tissue accumulation of EB was not influenced by experimental manipulation. Net fluid transport was measured in experiments exposing the intestinal mucosa to a high toxin concentration. Fluid secretion caused by the toxin was significantly attenuated by intravenous hexamethonium whereas no effect was observed after administration of nifedipine or granisetron. CONCLUSIONS: At a low toxin concentration, intramural reflexes are involved in the inflammatory response whereas axon reflexes contribute to tissue damage. At a high toxin concentration no nervous involvement in the toxin A response was demonstrated except for fluid secretion evoked by the toxin. PMID- 11413112 TI - Low socioeconomic class is a risk factor for upper and lower gastrointestinal symptoms: a population based study in 15 000 Australian adults. AB - BACKGROUND: The association of social class with health has been extensively studied, yet relationships between social class and gastrointestinal symptoms remain almost unexplored. AIMS: To examine relationships between social class and gastrointestinal symptoms in a population sample. METHODS: The prevalence of 16 troublesome gastrointestinal symptoms was determined by a postal questionnaire sent to 15 000 subjects (response rate 60%) and compared with a validated composite measure of socioeconomic status (index of relative socioeconomic disadvantage). Comparisons across social class were explored for five symptom categories (oesophageal symptoms; upper dysmotility symptoms; bowel symptoms; diarrhoea; and constipation). Results are reported as age standardised rate ratios with the most advantaged social class as the reference category. RESULTS: There were clear trends for the prevalence rates of all gastrointestinal symptoms to increase with decreasing social class. These trends were particularly strong for the five symptom categories. Lower social class was associated with a significantly (p<0.0001) higher number of symptoms reported overall and with a higher proportion of individuals reporting 1-2 symptoms and more than five symptoms. In both sexes, the most pronounced effects for subjects in the lowest social class were found for constipation (males: rate ratio 1.83 (95% confidence intervals (CI) 1.16-2.51); females: rate ratio 1.68 (95% CI 1.31-2.04)) and upper dysmotility symptoms (males: rate ratio 1.45 (95% CI 1.02-1.88); females: rate ratio 1.35 (95% CI 1.07-1.63)). Oesophageal symptoms and diarrhoea were not associated with social class. CONCLUSIONS: Troublesome gastrointestinal symptoms are linked to socioeconomic status with more symptoms reported by subjects in low socioeconomic classes. Low socioeconomic class should be considered a risk factor for both upper and lower gastrointestinal symptoms. PMID- 11413113 TI - LI-cadherin: a marker of gastric metaplasia and neoplasia. AB - BACKGROUND: Intestinal metaplasia is considered a risk factor for the development of gastric adenocarcinomas of the intestinal type and is found in approximately 20% of gastric biopsies. Conventional histology only detects advanced stages of intestinal metaplasia. AIMS: To study expression of the enterocyte specific adhesion molecule liver-intestinal (LI)-cadherin in intestinal metaplasia as well as in gastric cancer, and to evaluate its use as a diagnostic marker molecule. PATIENTS: Gastric biopsies (n=77) from 30 consecutive patients (n=30; aged 28-90 years) as well as surgically resected tissue samples (n=24) of all types of gastric carcinomas were analysed. METHODS: Single and double label immunofluorescence detection on cryosections of gastric biopsies; alkaline phosphatase antialkaline phosphatase method on paraffin embedded carcinoma tissue sections. RESULTS: Of 77 biopsies (from 30 patients), 12 (from 10 patients) stained positive for LI-cadherin. LI-cadherin staining correlated with the presence of intestinal metaplasia. Conventional histological diagnosis however failed to detect subtle gastric intestinal metaplasia (three of 10 patients). In contrast, only LI-cadherin and villin were positive in these cases whereas sucrase-isomaltase also failed to detect intestinal metaplasia in four of 10 patients. Well differentiated gastric carcinomas showed intense staining for LI cadherin while undifferentiated carcinomas showed only weak diffuse cytoplasmic staining. CONCLUSIONS: To detect early metaplastic changes in the gastric mucosa, LI-cadherin has a sensitivity superior to sucrase-isomaltase and conventional histology and comparable with that of villin. Its specificity exceeds that of villin. Thus LI-cadherin represents a new, reliable, and powerful marker molecule for early detection of gastric intestinal metaplasia and well differentiated adenocarcinomas. PMID- 11413114 TI - Genomic homogeneity in fibrolamellar carcinomas. AB - BACKGROUND: Fibrolamellar carcinoma (FLC) is a variant of hepatocellular carcinoma (HCC) with distinctive clinical and histological features. To date there have been few studies on the genotypic aspects of FLC and no previous attempts have been made to use the arbitrarily primed-polymerase chain reaction (AP-PCR) technique to detect genetic alterations in this disease. AIM: The aim of this study was to assess the degree of genomic heterogeneity of FLC using the AP PCR technique. METHODS: A total of 50 tissue samples of primary and metastatic FLCs from seven patients were microdissected. AP-PCR amplification of each genomic DNA sample was carried out using two arbitrary primers. RESULTS: DNA fingerprints of the primary FLCs and all their metastatic lesions (both synchronous and metachronous disease) were identical in an individual patient. The fingerprints were different between tumours of different patients. No evidence of intratumour heterogeneity was observed. CONCLUSIONS: Such genomic homogeneity in FLCs may explain their indolent growth. The absence of clonal evolution, which is present in other tumours (particularly HCCs), may explain the distinct behaviour in this tumour. The tumorigenic pathway and degree of somatic genomic changes in this disease may be less complex than in HCC. PMID- 11413115 TI - Coffee consumption and risk of colorectal cancer in a population based prospective cohort of Swedish women. AB - BACKGROUND: The presumed protective effect of coffee consumption on colorectal cancer, which is supported by case control studies, has not been confirmed in prospective cohort studies. Cohort studies are few in number however and often suffer from a small number of cases, limited attention to confounding variables, and a low percentage of heavy coffee drinkers. METHODS: We examined data from a large population based cohort of Swedish women who were free from cancer at the start of follow up, with a wide range of coffee consumption, information on many potentially confounding variables, and a larger number of cases than any previous cohort study of coffee consumption and colorectal cancer. RESULTS: During an average of 9.6 years of follow up of 61 463 women aged 40-74 years, we observed 460 incident cases of colorectal cancer (291 with colon cancer, 159 with rectal cancer, 10 with cancer at both sites). We found no association between coffee consumption and colorectal cancer risk. The risk ratio for drinking four or more cups per day compared with none was 1.04 (95% confidence interval 0.63-1.69; p for trend 0.84). The findings were similar for cancers of the distal and proximal colon and rectum. CONCLUSIONS: The recently published affirmative conclusions regarding the protective effect of coffee consumption may be premature. For patients seeking advice about coffee consumption, the evidence suggests that moderate or even high consumption will probably not influence the risk of colorectal cancer. PMID- 11413116 TI - Risks, costs, and compliance limit colorectal adenoma surveillance: lessons from a randomised trial. AB - BACKGROUND AND AIMS: In the USA and many other countries, endoscopic surveillance of colorectal adenoma patients is now widely practised. However, the optimal frequency and mode of such surveillance are not yet established. The aim of this trial was to compare surveillance at one, two, or five year intervals using either flexible sigmoidoscopy or colonoscopy. METHODS: Analysis of a randomised trial of flexible sigmoidoscopy and colonoscopy over one, two, or five years after stratification for "high" or "low" risk of recurrent adenomas. The trial started in 1984. RESULTS: A total of 776 patients were stratified into "high" (n=307) and "low" (n=469) recurrence risk groups and randomised to flexible sigmoidoscopy or colonoscopy at varying intervals. Only 81 recurrent adenomas (30/81 were >1 cm in diameter) were detected in the 2307 person years of follow up within the surveillance study. Adenoma recurrence was significantly higher in the high risk group (relative rate 1.82; 95% confidence interval 1.2-2.9) but recurrence rates per 1000 person years were low and not significantly different in those surveyed by colonoscopy or flexible sigmoidoscopy. Loss to follow up was greatest in those having an annual examination compared with two or five yearly surveillance examinations. Despite surveillance, invasive cancer developed in four patients compared with an expected value of 9.12 for the general population in England (p=0.10); of these four patients who developed cancers, only one was detected by surveillance examination. CONCLUSIONS: Adenoma recurrence rates were much lower than expected in both high and low risk groups. This suggests that endoscopic surveillance should be targeted at high risk groups. A surveillance interval of five years was as effective as shorter intervals in terms of cancer prevention, and was associated with similar compliance to two yearly examinations. PMID- 11413117 TI - Effects of cellular redox balance on induction of apoptosis by eicosapentaenoic acid in HT29 colorectal adenocarcinoma cells and rat colon in vivo. AB - BACKGROUND AND AIMS: Epidemiological evidence suggests n-3 polyunsaturated lipids may protect against colorectal neoplasia. Consumption of fish oil modulates crypt cytokinetics in humans, and crypt apoptosis in animal models. To explore these effects, we investigated involvement of caspase enzymes and cellular redox balance in the induction of apoptosis by eicosapentaenoic acid (EPA) in HT29 cells, and in rat colon in vivo. METHODS: Survival of HT29 cells grown with EPA in the presence of caspase inhibitors, antioxidants, or buthionine sulphoximine, an inhibitor of glutathione neosynthesis, was determined. The effects of EPA enriched fish oil and glutathione depletion on apoptosis in rat colon were assessed using microdissected crypts. RESULTS: Treatment of HT29 cells with EPA reduced viable cell number and activated caspase 3, prior to cell detachment. Antioxidants and caspase inhibitors blocked HT29 cell death whereas glutathione depletion increased it. Rats fed fish oil had higher crypt cell apoptosis than those fed corn oil, and glutathione depletion enhanced this effect. CONCLUSIONS: Incorporation of EPA into colonic epithelial cell lipids increases apoptosis. The results of this study, using both an animal and cell line model, support the hypothesis that this effect is mediated via cellular redox tone, and is sensitive to glutathione metabolism. The data suggest a mechanism whereby polyunsaturated fatty acids may influence the susceptibility of colorectal crypt cells to induction or progression of neoplasia. PMID- 11413118 TI - Gene expression of interleukin 18 in unstimulated peripheral blood mononuclear cells of patients with alcoholic cirrhosis. AB - BACKGROUND: Most patients with alcohol induced cirrhosis (AC) and chronic endotoxinaemia are not suffering from clinically evident systemic inflammatory reactions. This may be due to altered gene expression of cytokines, possibly related to endotoxin (for example, tolerance and sensitisation). Interleukin 18 (IL-18; interleukin gamma inducing factor) modulates local cytokine production in response to endotoxin (lipopolysaccharide (LPS)). AIM: To investigate the systemic immune response of patients with AC and to see if unstimulated peripheral blood mononuclear cells (PBMC) from patients with AC are activated and contribute to gene expression of IL-18. METHODS: Plasma levels of endotoxin (LPS) and serum levels of IL-18 were measured by enzyme linked immunoassay and the amoebocyte lysate test in 74 abstinent patients with different stages of AC (Child-Pugh stage A, n=18; B, n=22; C, n=34) and compared with healthy controls (n=43). Gene expression of IL-18 was assessed by semiquantitative reverse transcription-polymerase chain reaction in freshly isolated unstimulated PBMC of a subgroup of 14 patients with AC compared with five healthy controls. RESULTS: Gene expression of IL-18 specific mRNA in unstimulated PBMC was significantly enhanced in patients with advanced AC (Child-Pugh stage C) and correlated with plasma LPS and serum CD14 levels (Spearman rank correlation factors r=0.76 and r=0.72). Serum concentrations of IL-18 were also elevated compared with healthy controls (p<0.001) but correlation with serum levels of CD14 and plasma levels of LPS was much weaker compared with mRNA data (Spearman rank correlation factors r=0.47 and r=0.26). CONCLUSIONS: Our in vivo data suggest a presensitisation of "unstimulated" PBMC in the circulation of patients with AC by endotoxin. The term "unstimulated" may be inadequate in patients with AC. Further investigations are needed to define the exact mechanisms and localisation of sensitisation of PBMC in vivo. PMID- 11413119 TI - High arterial compliance in cirrhosis is related to low adrenaline and elevated circulating calcitonin gene related peptide but not to activated vasoconstrictor systems. AB - BACKGROUND AND AIMS: Static and dynamic functions of the wall of large arteries are largely unknown in cirrhosis in vivo. The present study was undertaken to determine arterial compliance (COMP(art)) in relation to vasodilator and vasoconstrictor systems in patients with cirrhosis. In addition, vasoactivity was manipulated by inhalation of oxygen. STUDY POPULATION AND METHODS: In 20 patients with alcoholic cirrhosis and 12 controls we determined COMP(art) (stroke volume relative to pulse pressure), cardiac output, plasma volume, systemic vascular resistance, central circulation time, plasma catecholamines, renin activity, endothelin-1, and calcitonin gene related peptide (CGRP) at baseline and during oxygen inhalation. RESULTS: COMP(art) was significantly increased in cirrhotic patients compared with controls (1.32 v 1.06 ml/mm Hg; p< 0.05) and inversely related to plasma adrenaline levels (r=-0.53; p<0.02) but positively related to circulating levels of CGRP (r=0.58; p<0.01). No significant relation was found for plasma noradrenaline, renin activity, or endothelin-1. COMP(art) was positively related to plasma volume (r=0.50; p<0.02) and inversely to systemic vascular resistance (r=-0.69; p<0.001) and central circulation time (r=-0.49; p<0.02). During oxygen inhalation, COMP(art) decreased (-13%; p<0.005) and systemic vascular resistance increased (+10%; p<0.001) towards normal values without significant changes in mean arterial pressure. Plasma adrenaline (-16%; p<0.01) decreased and the relation to COMP(art) disappeared. The relation of COMP(art) to CGRP and circulatory variables remained unchanged. CONCLUSION: Elevated arterial compliance in cirrhosis is related to low adrenaline, high CGRP, and systemic hyperdynamics but not to indicators of the activated vasoconstrictor systems (noradrenaline, renin, endothelin-1). Thus the altered static and dynamic characteristics of the wall of large arteries are intimately associated with circulatory and vasodilatory derangement in cirrhosis but biomanipulation indicates that the changes are, at least in part, reversible during isobaric conditions. PMID- 11413120 TI - After portal branch ligation in the rat, cellular proliferation is associated with selective induction of c-Ha-ras, p53, cyclin E, and Cdk2. AB - BACKGROUND: In liver regeneration after portal branch ligation we previously showed that early cellular changes are observed in both the proliferating and atrophying liver lobes. They are therefore not indicative of future proliferative response. In this study we attempted to define precisely, in the same model, the time at which the cellular processes diverge between the lobes by measuring various parameters associated with cellular proliferation. We also investigated the possible role of inhibitors of cell proliferation in the absence of progression towards the S phase in the atrophying lobes. AIMS: Expression of p53, c-Ha-ras, cyclin E, cyclin dependent kinase (Cdk2), transforming growth factor (TGF)-beta, and interleukin (IL)-1alpha and IL-1beta were assessed in relation to their potential role in proliferating and atrophying cellular phenomenons. METHODS: Immunohistochemistry, northern blotting, western blotting, and reverse transcription-polymerase chain reaction were performed, mainly at time points corresponding to mid-G1/S phase progression (8-24 hours after surgery). RESULTS: The common and thus most likely non-specific response was still evident 5-8 hours after surgery and included an increase in IL-1 mRNA as well as p53 and cyclin E proteins. From 12 hours onwards, p53, c-Ha-ras, cyclin E, and Cdk2 were selectively induced in proliferating lobes whereas IL-1beta was predominantly activated in atrophying lobes. No changes in TGF-beta or IL-1alpha expression were observed at the same time points in any of the liver lobes. CONCLUSIONS: The initial response to portal branch ligation and thus probably to partial hepatectomy seems to be non-specific for at least eight hours. Thereafter, p53, c Ha-ras, cyclin E, and Cdk2 seem to drive cellular proliferation while IL-1beta is associated with cellular atrophy. In contrast, TGF-beta and IL-1alpha do not seem to play a role in determining the commitment of cells towards atrophy or proliferation. PMID- 11413121 TI - Tumour necrosis factor alpha impairs function of liver derived T lymphocytes and natural killer cells in patients with primary sclerosing cholangitis. AB - BACKGROUND: Primary sclerosing cholangitis (PSC) is considered to be a chronic autoimmune disease where infiltrating T lymphocytes have been implicated in the destruction of bile ducts. Altered function of these T cells may reflect abnormalities in the immune response leading to tissue damage. AIM: We investigated the proliferative and functional capacity of freshly isolated liver derived T lymphocytes (LDLs) and natural killer (NK) cells from PSC patients. METHODS: The proliferative responses to common mitogens such as phytohaemagglutinin (PHA), concanavalin A (Con A), and lipopolysaccharide (LPS) were studied, and the cytotoxic function of T lymphocytes was measured using allogeneic target cells. NK (CD56(+)/16(+)) cytotoxic function was measured using the two cell lines K562 (NK sensitive) and Raji lymphoma cells (NK resistant). RESULTS: Compared with patients with primary biliary cirrhosis (PBC), autoimmune hepatitis (AIH), and normal controls (without liver disease), in PSC: (1) LDLs contained a low percentage of T cells; (2) there was significantly decreased expression of interleukin (IL)-2 receptor (p<0.001) on activated T cells (HLA DR(+)); (3) LDLs but not peripheral blood lymphocytes had significantly impaired proliferative responses to mitogens such as PHA, Con A, and LPS (p< 0.001); (4) no cytotoxic activity of PSC liver T and NK cells was recorded; (5) significantly higher levels of tumour necrosis factor alpha (TNF-alpha) and IL-1beta but lower levels of IL-2, IL-10, and interferon gamma were found in the supernatants of mitogen stimulated LDL cultures (p<0.001); (6) higher percentages of freshly isolated PSC LDLs contained intracytoplasmic TNF-alpha and IL-1beta; and (7) pretreatment of PSC LDLs in vitro with neutralising TNF antibodies significantly enhanced proliferative responses and allowed IL-2 receptor expression following stimulation. In addition, the impaired cytolytic activity of both NK and T cells was partially restored. Impaired proliferative or functional capacity of liver derived T cells was not observed in either PBC or AIH patients. CONCLUSIONS: We suggest that reduced T cell reactivity in liver infiltrating cells obtained from patients with PSC is due to high local production of TNF-alpha. Our findings indicate that the use of anti-TNF antibodies as an alternative treatment for PSC patients should be evaluated. PMID- 11413122 TI - Pathological documentation of complete elimination of Barrett's metaplasia following endoscopic multipolar electrocoagulation therapy. AB - The previous paradigm that Barrett's is an irreversible premalignant lesion has recently been challenged by a proliferation of reports documenting elimination of Barrett's by a variety of endoscopic techniques. Whether Barrett's is entirely eliminated is unknown as endoscopic biopsy samples the surface of the epithelium only. Numerous reports document underlying specialised columnar epithelium in many of these trials. Until now there have been no reports of pathological examination of the entire oesophagus as a specimen. This case documents complete elimination of intestinal metaplasia from the oesophagus and supports the biological plausibility of these research techniques. PMID- 11413123 TI - Classification of oesophageal motility abnormalities. AB - Manometric examination of the oesophagus frequently reveals abnormalities whose cause is unknown and whose physiological importance is not clear. A large body of literature dealing with oesophageal motility abnormalities has evolved over the past few decades but comparisons among studies have been compromised by the lack of a widely accepted system for classifying the abnormal motility patterns, and by the lack of uniform diagnostic criteria for the putative disorders. Based on an extensive review and analysis of the literature, this report suggests an operational scheme to be used for the general classification of oesophageal motility abnormalities, and proposes standardised manometric criteria for the putative oesophageal motility disorders. By applying the guidelines proposed in this report, clinicians and researchers can determine if their patients fulfil the manometric criteria for a putative motility disorder. This should facilitate and improve comparisons among patients and studies. However, it is important to emphasise that fulfilment of the proposed criteria does not establish the clinical importance of the motility abnormalities. PMID- 11413124 TI - Recurrent Clostridium difficile diarrhoea. PMID- 11413126 TI - Identification of novel bifunctional calmodulin-binding and microtubule stabilizing motifs in STOP proteins. AB - Although microtubules are intrinsically labile tubulin assemblies, many cell types contain stable polymers, resisting depolymerizing conditions such as exposure to the cold or the drug nocodazole. This microtubule stabilization is largely due to polymer association with STOP proteins. There are several STOP variants, some with capacity to induce microtubule resistance to both the cold and nocodazole, others with microtubule cold stabilizing activity only. These microtubule-stabilizing effects of STOP proteins are inhibited by calmodulin and we now demonstrate that they are determined by two distinct kinds of repeated modular sequences (Mn and Mc), both containing a calmodulin-binding peptide, but displaying different microtubule stabilizing activities. Mn modules induce microtubule resistance to both the cold and nocodazole when expressed in cells. Mc modules, which correspond to the STOP central repeats, have microtubule cold stabilizing activity only. Mouse neuronal STOPs, which induce both cold and drug resistance in cellular microtubules, contain three Mn modules and four Mc modules. Compared with neuronal STOPs, the non-neuronal F-STOP lacks multiple Mn modules and this corresponds with an inability to induce nocodazole resistance. STOP modules represent novel bifunctional calmodulin-binding and microtubule stabilizing sequences that may be essential for the generation of the different patterns of microtubule stabilization observed in cells. PMID- 11413125 TI - Clinical guidelines on the management of hepatitis C. PMID- 11413127 TI - Effect of osmolytes and chaperone-like action of P-protein on folding of nucleocapsid protein of Chandipura virus. AB - Amino acid sequences of nucleocapsid proteins are mostly conserved among different rhabdoviruses. The protein plays a common functional role in different RNA viruses by enwrapping the viral genomic RNA in an RNase-resistant form. Upon expression of the nucleocapsid protein alone in COS cells and in bacteria, it forms large insoluble aggregates. In this work, we have reported for the first time the full-length cloning of the N gene of Chandipura virus and its expression in Escherichia coli in a soluble monomeric form and purification using nonionic detergents. The biological activity of the soluble recombinant protein has been tested, and it was found to possess efficient RNA-binding ability. The state of aggregation of the recombinant protein was monitored using light scattering. In the absence of nonionic detergents, it formed large aggregates. Aggregation was significantly reduced in the presence of osmolytes such as d-sorbitol. Aggregate formation was suppressed in the presence of another viral product, phosphoprotein P, in a chaperone-like manner. Both the osmolyte and phosphoprotein P also suppressed aggregation to a great extent during refolding from a guanidine hydrochloride-denatured form. The function of the phosphoprotein and osmolyte appears to be synergistic to keep the N-protein in a soluble biologically competent form in virus-infected cells. PMID- 11413128 TI - Synergistic movements of Ca(2+) and Bax in cells undergoing apoptosis. AB - Apoptosis is a physiological counterbalance to mitosis and plays important roles in tissue development and homeostasis. Cytosolic Ca(2+) has been implicated as a proapoptotic second messenger involved in both triggering apoptosis and regulating cell death-specific enzymes. A critical early event in apoptosis is associated with the redistribution of Bax from cytosol to mitochondria and endoplasmic reticulum (ER) membranes; however, the molecular mechanism of Bax translocation and its relationship to Ca(2+) is largely unknown. Here we provide functional evidence for a synergistic interaction between the movements of intracellular Ca(2+) and cytosolic Bax in the induction of apoptosis. Overexpression of Bax in cultured cells causes a loss of ER Ca(2+) content. Depletion of ER Ca(2+) through activation of the ryanodine receptor enhances the participation of Bax into the mitochondrial membrane. Neither Bax translocation nor Bax-induced apoptosis is affected by buffering of cytosolic Ca(2+) with 1,2 bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, suggesting that depletion of ER Ca(2+) rather than elevation of cytosolic Ca(2+) is the signal for cell apoptosis. This dynamic interplay of Ca(2+) and Bax movements may serve as an amplifying factor in the initial signaling steps of apoptosis. PMID- 11413129 TI - Proteinase-activated receptor-2-mediated activation of stress-activated protein kinases and inhibitory kappa B kinases in NCTC 2544 keratinocytes. AB - In this study we examined the regulation of the stress-activated protein (SAP) kinases and inhibitory kappa B kinases (IKKs) through stimulation of the novel G protein-coupled receptor proteinase-activated receptor-2 in the human keratinocyte cell line NCTC2544. Trypsin and the peptide SLIGKV stimulated a time dependent increase in both c-Jun N-terminal kinase and p38 mitogen-activated protein kinase activity. Trypsin also stimulated NF kappa B-DNA binding and the activation of the upstream kinases IKK alpha and -beta. Phorbol 12-myristate 13 acetate also strongly activated both SAP kinases and IKK isoforms, suggesting the potential for a protein kinase C-mediated regulatory mechanism underlying the effects of trypsin. Pre-incubation with selective protein kinase C (PKC) inhibitors GF109203X and Go6983, or transfection of dominant negative (DN)-PKC alpha, abolished phorbol 12-myristate 13-acetate-mediated c-Jun N-terminal kinase activity, although it only partially inhibited the response to trypsin. In contrast, Go6983 reduced trypsin-stimulated p38 mitogen-activated protein kinase activity to a greater extent than GF109203X, although DN-PKC alpha or PKC zeta had no substantial effect. Additionally, inhibitors of PKC partially reduced trypsin-stimulated IKK alpha activity but abolished that of IKK beta, whereas DN PKC alpha but not DN-PKC zeta substantially reduced trypsin-stimulated Flag-IKK beta activity. This study shows for the first time proteinase-activated receptor 2-mediated stimulation of both SAP kinase and IKK signaling and differing roles for PKC isoforms in the regulation of each pathway. PMID- 11413130 TI - Cytoskeletal changes regulated by the PAK4 serine/threonine kinase are mediated by LIM kinase 1 and cofilin. AB - PAK4 is the most recently identified member of the PAK family of serine/threonine kinases. PAK4 differs from other members of the PAK family in sequence and in many of its functions. Previously, we have shown that an important function of this kinase is to mediate the induction of filopodia in response to the Rho GTPase Cdc42. Here we show that PAK4 also regulates the activity of the protein kinase LIM kinase 1 (LIMK1). PAK4 was shown to interact specifically with LIMK1 in binding assays. Immune complex kinase assays revealed that both wild-type and constitutively active PAK4 phosphorylated LIMK1 even more strongly than PAK1, and activated PAK4 stimulated LIMK1's ability to phosphorylate cofilin. Immunofluorescence experiments revealed that PAK4 and LIMK1 cooperate to induce cytoskeletal changes in C2C12 cells. Furthermore, dominant negative LIMK1 and a mutant cofilin inhibited the specific cytoskeletal and cell shape changes that were induced in response to a recently characterized constitutively activated PAK4 mutant. PMID- 11413131 TI - Recognition of E-cadherin by integrin alpha(E)beta(7): requirement for cadherin dimerization and implications for cadherin and integrin function. AB - We have investigated the importance of dimerization of E-cadherin in the heterophilic adhesive interaction between E-cadherin and integrin alpha(E)beta(7). Dimerization of cadherin molecules in parallel alignment is known to be essential for homophilic adhesion and has been attributed to Ca(2+) dependent interactions in the domain 1-2 junction or to cross-intercalation of Trp2 from one molecule to the other. We have disrupted either or both of these proposed mechanisms by point mutations in E-cadherin-Fc and have tested the modified proteins for alpha(E)beta(7)-mediated cell adhesion. Prevention of Trp2 intercalation had no adverse effect on integrin-mediated adhesion, whereas disruption of Ca(2+) binding permitted adhesion but with reduced efficiency. Both modifications in combination abolished recognition by alpha(E)beta(7). In EGTA, alpha(E)beta(7) adhered to wild type E-cadherin but not to the Trp2 deletion mutant. Independent evidence that the mutations prevented either or both mechanisms for dimerization is presented. The data show that dimerization is required for recognition by alpha(E)beta(7) and that it can take place by either of two mechanisms. Implications for the roles of the alpha(E) and beta(7) integrin subunits in ligand binding and for Trp2 and Ca(2+) in the assembly of cadherin complexes are discussed. PMID- 11413132 TI - A new insight into metallothionein (MT) classification and evolution. The in vivo and in vitro metal binding features of Homarus americanus recombinant MT. AB - We report the synthesis and characterization of a Homarus americanus MT-cDNA (MTH) through retrotranscription of MTH-mRNA from metal-injected lobsters. Heterologous Escherichia coli expression in zinc- and copper-supplemented medium was achieved for MTH, the two domains betabetaMTH and betaalphaMTH and three site directed mutants, betabetaC9H, betaalphaC37H, and betaalphaE31C/T34C. The in vivo conformed metal complexes and the in vitro substituted cadmium aggregates were characterized. Major stoichiometries of M(II)6-MTH for the entire MTH and M(II)3 betabetaMTH and M(II)3-betaalphaMTH for the independent domains fully validated our expression system. A low affinity binding site for a seventh Zn(II) in the in vivo synthesized MTH was located in the betaalpha domain. Additionally, minor M(II)4 species were found for each domain. Both single Cys to His mutations exhibited a similar reduction of their in vivo zinc binding ability but differed in their cadmium binding behavior when compared with the wild-type forms. Conversely, the double mutant showed an enhanced zinc and cadmium binding capacity. In vivo synthesis of MTH and of its independent domains in the presence of copper only afforded heterometallic copper-zinc species. These findings allow consideration of MTH as a zinc thionein and question the view of all crustacea MT structures as copper thioneins. Furthermore, a new approach for the evolutionary and functional classification of MT is proposed, based on the stoichiometry of metal-MT species and molecular phylogenetic analysis. PMID- 11413133 TI - Purinergic-independent calcium signaling mediates recovery from hepatocellular swelling: implications for volume regulation. AB - Swelling of hepatocytes and other epithelia activates volume-sensitive ion channels that facilitate fluid and electrolyte efflux to restore cell volume, but the responsible signaling pathways are incompletely defined. Previous work in model HTC rat hepatoma cells has indicated that swelling elicits ATP release, which stimulates P2 receptors and activates Cl(-) channels, and that this mechanism is essential for hepatocellular volume recovery. Since P2 receptors are generally coupled to Ca(2+) signaling pathways, we determined whether hepatocellular swelling affected cytosolic [Ca(2+)], and if this involved a purinergic mechanism. Exposure of HTC cells to hypotonic media evoked an increase in cytosolic [Ca(2+)], which was followed by activation of K(+) and Cl(-) currents. Maneuvers that interfered with swelling-induced increases in cytosolic [Ca(2+)], including extracellular Ca(2+) removal and intracellular Ca(2+) store depletion with thapsigargin, inhibited activation of membrane currents and volume recovery. However, the swelling-induced increases in cytosolic [Ca(2+)] were unaffected by either extracellular ATP depletion with apyrase or blockade of P2 receptors with suramin. These findings indicate that swelling elicits an increase in hepatocellular Ca(2+), which is essential for ion channel activation and volume recovery, but that this increase does not stem from activation of volume sensitive P2 receptors. Collectively, these observations imply that regulatory responses to hepatocellular swelling involve a dual requirement for a purinergic independent Ca(2+) signaling cascade and a Ca(2+)-independent purinergic signaling pathway. PMID- 11413134 TI - Sarcolipin, the shorter homologue of phospholamban, forms oligomeric structures in detergent micelles and in liposomes. AB - The human 31-amino acid integral membrane protein sarcolipin (SLN), which regulates the sarcoplasmic reticulum Ca-ATPase in fast-twitch skeletal muscle, was chemically synthesized. Appropriate synthesis and purification strategies were used to achieve high purity and satisfactory yields of this hydrophobic and poorly soluble protein. Structural and functional properties of SLN were analyzed and compared with the homologous region of human phospholamban (PLB) comprising residues Ala(24)-Leu(52) (PLB-(24-52)), the regulatory protein of the cardiac sarcoplasmic reticulum Ca-ATPase. Circular dichroism spectroscopy showed that SLN is a predominantly alpha-helical protein and that the secondary structure is highly resistant to SDS and thermal denaturation. In this respect SLN is remarkably similar to PLB-(24-52). However, SLN is monomeric in SDS gels, whereas PLB-(24-52) shows a monomer-pentamer equilibrium typical for native PLB. Analytical ultracentrifugation experiments revealed that SLN oligomerizes in the presence of the nonionic detergents octylpolyoxyethylene and octyl glucoside in a concentration-dependent manner. No plateau was observed, and a pentameric state was only reached at much higher protein concentrations compared with PLB-(24-52). Chemical cross-linking showed that also in liposomes SLN has the ability to self associate to oligomers. PLB-(24-52) specifically oligomerized to pentamers in the presence of octylpolyoxyethylene as well as in liposomes at low protein concentrations. In the presence of octylpolyoxyethylene pentamers were the main oligomeric species, whereas in liposomes monomers and dimers were predominant. Increasing the protein concentration led to self-association of PLB-(24-52) pentamers in the presence of octylpolyoxyethylene. Functional reconstitution of Ca-ATPase with PLB-(24-52) and SLN in liposomes showed that both proteins regulate the Ca-ATPase in a similar manner. PMID- 11413135 TI - Regulation of L-type calcium channels in pituitary GH(4)C(1) cells by depolarization. AB - The neurosecretory anterior pituitary GH(4)C(1) cells exhibit the high voltage activated dihydropyridine-sensitive L-type and the low voltage-activated T-type calcium currents. The activity of L-type calcium channels is tightly coupled to secretion of prolactin and other hormones in these cells. Depolarization induced by elevated extracellular K(+) reduces the dihydropyridine (+)-[(3)H]PN200-110 binding site density and (45)Ca(2+) uptake in these cells (). This study presents a functional analysis by electrophysiological techniques of short term regulation of L-type Ca(2+) channels in GH(4)C(1) cells by membrane depolarization. Depolarization of GH(4)C(1) cells by 50 mm K(+) rapidly reduced the barium currents through L-type calcium channels by approximately 70% and shifted the voltage dependence of activation by 10 mV to more depolarized potentials. Down regulation depended on the strength of the depolarizing stimuli and was reversible. The currents recovered to near control levels on repolarization. Down regulation of the calcium channel currents was calcium-dependent but may not have been due to excessive accumulation of intracellular calcium. Membrane depolarization by voltage clamping and by veratridine also produced a down regulation of calcium channel currents. The down-regulation of the currents had an autocrine component. This study reveals a calcium-dependent down-regulation of the L-type calcium channel currents by depolarization. PMID- 11413136 TI - Self-association of class II transactivator correlates with its intracellular localization and transactivation. AB - Class II transactivator (CIITA) is the master regulator of major histocompatibility complex class II genes that regulates both B lymphocyte specific and interferon gamma-inducible expression. Here we identify protein regions and examine mechanisms that determine the intracellular distribution of CIITA. We show that two separate regions of CIITA mediate nuclear export: amino acids 1-114 and 408-550. Both regions interact with the export receptor CRM-1. The CIITA region spanning amino acids 408-550 of CIITA also determines its ability for homotypic self-association as well as heterotypic interactions with other regions residing at the amino and carboxyl termini of the protein. These observations are in line with data demonstrating that co-expression of amino- and carboxyl-terminal parts of CIITA promote subcellular relocalization and, remarkably, rescue transcriptional activation by individually inert molecules. CIITA point mutations that impair nuclear import and abolish its activation function show reduced self-association. We propose that the concerted action of homo- and heterotypic interactions of CIITA determine proper protein configuration that in turn controls its nucleocytoplasmic trafficking. PMID- 11413137 TI - Characterization of selected strains of pneumococcal surface protein A. AB - Several proteins, in addition to the polysaccharide capsule, have recently been implicated in the full virulence of the Streptococcus pneumoniae bacterial pathogen. One of these novel virulence factors of S. pneumoniae is pneumococcal surface protein A (PspA). The N-terminal, cell surface exposed, and functional part of PspA is essential for full pneumococcal virulence, as evidenced by the fact that antibodies raised against this part of the protein are protective against pneumococcal infections. PspA has recently been implicated in anti complementary function as it reduces complement-mediated clearance and phagocytosis of pneumococci. Several recombinant N-terminal fragments of PspA from different strains of pneumococci, Rx1, BG9739, BG6380, EF3296, and EF5668, were analyzed using circular dichroism, analytical ultracentrifugation sedimentation velocity and equilibrium methods, and sequence homology. Uniformly, all strains of PspA molecules studied have a high alpha-helical secondary structure content and they adopt predominantly a coiled-coil structure with an elongated, likely rod-like shape. No beta-sheet structures were detected for any of the PspA molecules analyzed. All PspAs were found to be monomeric in solution with the exception of the BG9739 strain which had the propensity to partially aggregate but only into a tetrameric form. These structural properties were correlated with the functional, anti-complementary properties of PspA molecules based on the polar distribution of highly charged termini of its coiled-coil domain. The recombinant Rx1 PspA is currently under consideration for pneumococcal vaccine development. PMID- 11413138 TI - Heme-ligating histidines in flavocytochrome b(558): identification of specific histidines in gp91(phox). AB - The phagocyte NADPH-dependent oxidase generates superoxide (O(2)) by reducing molecular oxygen through flavocytochrome b(558) (flavocytochrome b), a heterodimeric oxidoreductase composed of gp91(phox) and p22(phox) subunits. Although each flavocytochrome b molecule contains two heme groups, their precise distribution within the heterodimer is unknown. Among functionally and/or structurally related oxidoreductases, histidines at codons 101, 111, 115, 119, 209, 210, and 222 of gp91(phox) are conserved and potential candidates to ligate heme. We compared biochemical and functional features of normal flavocytochrome b with those in cells expressing gp91(phox) harboring amino acid substitutions at each of these histidines. Surface expression of flavocytochrome b and heterodimer formation were relatively unaffected in cells expressing gp91(phox) H111L, H119L, or H210L. These mutations also had no effect on the flavocytochrome b heme spectrum, although NADPH oxidase activity was decreased in cells expressing gp91(phox) H119L or H210L. In contrast, gp65 was not processed to gp91(phox), heterodimers did not form, and flavocytochrome b was not expressed on the surface of cells expressing gp91(phox) H101L, H115L, H115D, H209C, H209Y, H222L, H222C, or H222R. Similarly, this subset of mutants lacked detectable O(2)-generating activity, and flavocytochrome b purified from these cells contained little or no heme. These findings demonstrate that His(101), His(115), His(209), and His(222) of gp91(phox) are critical for heme binding and biosynthetic maturation of flavocytochrome b. PMID- 11413139 TI - hPop5, a protein subunit of the human RNase MRP and RNase P endoribonucleases. AB - The RNase MRP and RNase P particles both function as endoribonucleases. RNase MRP has been implicated in the processing of precursor-rRNA, whereas RNase P has been shown to function in the processing of pre-tRNA. Both ribonucleoprotein particles have an RNA component that can be folded into a similar secondary structure and share several protein components. We have identified human, rat, mouse, cow, and Drosophila homologues of the Pop5p protein subunit of the yeast RNase MRP and RNase P complexes. The human Pop5 cDNA encodes a protein of 163 amino acids with a predicted molecular mass of 18.8 kDa. Polyclonal antibodies raised against recombinant hPop5 identified a 19-kDa polypeptide in HeLa cells and showed that hPop5 is associated with both RNase MRP and RNase P. Using affinity-purified anti hPop5 antibodies, we demonstrated that the endogenous hPop5 protein is localized in the nucleus and accumulates in the nucleolus, which is consistent with its association with RNase MRP and RNase P. Catalytically active RNase P was partially purified from HeLa cells, and hPop5 was shown to be associated with it. Finally, the evolutionarily conserved acidic C-terminal tail of hPop5 appeared to be required neither for complex formation nor for RNase P activity. PMID- 11413140 TI - Formation of highly reactive gamma-ketoaldehydes (neuroketals) as products of the neuroprostane pathway. AB - Neuroprostanes are prostaglandin-like compounds produced by free radical-induced peroxidation of docosahexaenoic acid, which is highly enriched in the brain. We previously described the formation of highly reactive gamma-ketoaldehydes (isoketals) as products of the isoprostane pathway of free radical-induced peroxidation of arachidonic acid. We therefore explored whether isoketal-like compounds (neuroketals) are also formed via the neuroprostane pathway. Utilizing mass spectrometric analyses, neuroketals were found to be formed in abundance in vitro during oxidation of docosahexaenoic acid and were formed in greater abundance than isoketals during co-oxidation of docosahexaenoic and arachidonic acid. Neuroketals were shown to rapidly adduct to lysine, forming lactam and Schiff base adducts. Neuroketal lysyl-lactam protein adducts were detected in nonoxidized rat brain synaptosomes at a level of 0.09 ng/mg of protein, which increased 19-fold following oxidation in vitro. Neuroketal lysyl-lactam protein adducts were also detected in vivo in normal human brain at a level of 9.9 +/- 3.7 ng/g of brain tissue. These studies identify a new class of highly reactive molecules that may participate in the formation of protein adducts and protein protein cross-links in neurodegenerative diseases and contribute to the injurious effects of other oxidative pathologies in the brain. PMID- 11413141 TI - Shigella invasion of macrophage requires the insertion of IpaC into the host plasma membrane. Functional analysis of IpaC. AB - Shigella infects residential macrophages via the M cell entry, after which the pathogen induces macrophage cell death. The bacterial strategy of macrophage infection, however, remains largely speculative. Wild type Shigella flexneri (YSH6000) invaded macrophages more efficiently than the noninvasive mutants, where YSH6000 induced large scale lamellipodial extension including ruffle formation around the bacteria. When macrophages were infected with the noninvasive ipaC mutant, the invasiveness and induction of membrane extension were dramatically reduced as compared with that of YSH6000. J774 macrophages infected with YSH6000 showed tyrosine phosphorylation of several proteins including paxillin and c-Cbl, and this pattern was distinctive from those stimulated by Salmonella typhimurium or phorbol ester. Upon addition of IpaC into the external medium of macrophages, membrane extensions were rapidly induced, and this promoted uptake of Escherichia coli. The exogenously added IpaC was found to be integrated into the host cell membrane as detected by immunostaining. The IpaC domain required for the induction of membrane extension from J774 was narrowed down within the region of residues 117-169, which contains a putative membrane spanning sequence. Our data indicate that Shigella directs its own entry into macrophages, and the IpaC domain which is required for the association with its host membrane is crucial. PMID- 11413142 TI - Identification and characterization of Harc, a novel Hsp90-associating relative of Cdc37. AB - Although little is known about the precise mechanisms by which the molecular chaperone Hsp90 recognizes its client proteins, Cdc37 has been shown to play a critical role in the targeting of Hsp90 to client protein kinases. Described here is the identification and characterization of a novel 35-kDa human protein that is 31% identical to Cdc37. We have named this novel protein Harc (Hsp90 associating relative of Cdc37). Northern blot analysis revealed the presence of Harc mRNA in several human tissues, including liver, skeletal muscle, and kidney. Biochemical fractionation and immunofluorescent localization of epitope-tagged Harc (i.e. FLAG-Harc) indicated that it is present in the cytoplasm of cells. FLAG-Harc binds Hsp90 but unlike Cdc37 does not bind Src family kinases or Raf-1. Mapping experiments indicate that the central 120 amino acids of both Harc and Cdc37 constitute a Hsp90-binding domain not described previously. FLAG-Harc is basally serine-phosphorylated and hyperphosphorylated when co-expressed with an activated mutant of the Src family kinase Hck. Notably, FLAG-Harc forms complexes with Hsp90, Hsp70, p60Hop, immunophilins, and an unidentified p22 protein but not with the Hsp90 co-chaperone p23. Thus Harc likely represents a novel participant in Hsp90-mediated protein folding, potentially targeting Hsp90 to Hsp70-client protein heterocomplexes. PMID- 11413143 TI - Overexpression of the myristoylated alanine-rich C-kinase substrate inhibits cell adhesion to extracellular matrix components. AB - Mice lacking the myristoylated alanine-rich C-kinase substrate, or MARCKS protein, exhibit abnormalities consistent with a defect in the ability of neurons to migrate appropriately during forebrain development. To investigate the possibility that this phenotype could be due to disruption of normal cellular adhesion to extracellular matrix, an assay was developed in which 293 cells co expressing MARCKS and green fluorescent protein were tested for their adhesion competence on various substrates. Fluorescence-activated cell sorting of adherent and non-adherent green fluorescent protein-expressing cells demonstrated that wild-type MARCKS inhibited adhesion of cells to fibronectin, whereas a non myristoylated mutant did not inhibit adhesion of cells to a variety of substrates. The fibronectin competitive inhibitor RGD peptide inhibited adhesion of cells expressing all MARCKS variants equally. Cytochalasin D inhibited the adhesion of cells expressing non-myristoylated MARCKS, but did not further decrease the adhesion of cells expressing adhesion-inhibitory proteins. Confocal microscopy demonstrated the presence of inhibitory, myristoylated MARCKS at the plasma membrane, suggesting that localization at this region might be important for MARCKS to inhibit cellular adhesion. These data suggest a possible myristoylation-dependent function of MARCKS to inhibit cellular adhesion to extracellular matrix proteins, indicating a potential mechanism for the cell migration defects seen in the MARCKS-deficient mice. PMID- 11413144 TI - Induction of secondary structure in a COOH-terminal peptide of histone H1 by interaction with the DNA: an infrared spectroscopy study. AB - We have studied the conformation of the peptide Ac-EPKRSVAFKKTKKEVKKVATPKK (CH 1), free in solution and bound to the DNA, by Fourier-transform infrared spectroscopy. The peptide belongs to the COOH-terminal domain of histone H1(0) (residues 99-121) and is adjacent to the central globular domain of the protein. In aqueous (D(2)O) solution the amide I' is dominated by component bands at 1643 cm(-1) and 1662 cm(-1), which have been assigned to random coil conformations and turns, respectively. In accordance with previous NMR results, the latter component has been interpreted as arising in turn-like conformations in rapid equilibrium with unfolded states. The peptide becomes fully structured either in 90% trifluoroethanol (TFE) solution or upon interaction with the DNA. In these conditions, the contributions of turn (1662 cm(-1)) and random coil components virtually disappear. In TFE, the spectrum is dominated by the alpha-helical component (1654 cm(-1)). The band at 1662 cm(-1) shifts to 1670 cm(-1), and has been assigned to the COOH-terminal TPKK motif in a more stable turn conformation. A band at 1637 cm(-1), also present in TFE, has been assigned to 3(10) helical structure. The amide I' band of the complexes with the DNA retains the components that were attributed to 3(10) helix and the TPKK turn. In the complexes with the DNA, the alpha-helical component observed in TFE splits into two components at 1657 cm(-1) and 1647 cm(-1). Both components are inside the spectral region of alpha-helical structures. Our results support the presence of inducible helical and turn elements, both sharing the character of DNA-binding motifs. PMID- 11413145 TI - Thrombin receptor signaling to cytoskeleton requires Hsp90. AB - Thrombin is a serine protease that evokes various cellular responses involved in injury and repair of the nervous system through the activation of protease activated receptor-1 (PAR-1). Signals that modulate cell morphology precede most PAR-1 effects, but the initial signal transduction molecules are not known. Using the yeast two-hybrid system, we identified Hsp90, a chaperone with known signaling properties, as a binding partner of PAR-1. The interaction was confirmed by glutathione S-transferase pull-down, overlay, and co immunoprecipitation assays. Morphological assays in mouse astrocytes were carried out to evaluate the importance of Hsp90 during cytoskeletal signaling. Reducing Hsp90 levels by antisense treatment or disruption of the Hsp90.PAR-1 complex by the Hsp90-specific drug geldanamycin attenuated thrombin-mediated astrocyte shape changes. Furthermore, overexpression of the PAR-1 cytoplasmic tail abrogated thrombin-induced cytoskeletal changes in neuronal cells. Treatment with geldanamycin specifically inhibited activation of RhoA without affecting thrombin mediated intracellular calcium release, revealing the regulation of a distinct signaling pathway by Hsp90. Taken together, these studies demonstrate that Hsp90 may be essential for PAR-1-mediated signaling to the cytoskeleton. PMID- 11413146 TI - The N-terminal and C-terminal domains of RAP1 are dispensable for chromatin opening and GCN4-mediated HIS4 activation in budding yeast. AB - Repressor activator protein 1 (RAP1) assists GCN4-mediated HIS4 activation by overcoming some repressive aspect of chromatin structure to facilitate GCN4 binding. RAP1 also participates in other nuclear processes, and discrete domains of RAP1 have been shown to have specific properties including DNA binding, DNA bending, transcriptional activation, and silencing and telomere functions. To investigate whether specific domains of RAP1 are required to "open" chromatin and help GCN4 to activate the HIS4 gene, we examined the abilities of different truncated RAP1 proteins to perturb positioned nucleosomes via a nucleosomal RAP1 site in a yeast episome in vivo, and we tested HIS4 activation in yeast strains harboring truncated RAP1 mutants. We found that neither the DNA bending domain nor the putative activation domain of RAP1 is required for its ability to perturb the chromatin structure of a plasmid containing a RAP1 site. Similarly, neither the putative activation domain nor the N-terminal DNA-bending domain was required for GCN4-mediated activation of HIS4. We also used a rap1(ts) mutant to show that continuous occupancy of the HIS4 promoter by RAP1 is required for GCN4-mediated gene activation. PMID- 11413147 TI - Involvement of the nucleotide excision repair protein UvrA in instability of CAG*CTG repeat sequences in Escherichia coli. AB - Several human genetic diseases have been associated with the genetic instability, specifically expansion, of trinucleotide repeat sequences such as (CTG)(n).(CAG)(n). Molecular models of repeat instability imply replication slippage and the formation of loops and imperfect hairpins in single strands. Subsequently, these loops or hairpins may be recognized and processed by DNA repair systems. To evaluate the potential role of nucleotide excision repair in repeat instability, we measured the rates of repeat deletion in wild type and excision repair-deficient Escherichia coli strains (using a genetic assay for deletions). The rate of triplet repeat deletion decreased in an E. coli strain deficient in the damage recognition protein UvrA. Moreover, loops containing 23 CTG repeats were less efficiently excised from heteroduplex plasmids after their transformation into the uvrA(-) strain. As a result, an increased proportion of plasmids containing the full-length repeat were recovered after the replication of heteroduplex plasmids containing unrepaired loops. In biochemical experiments, UvrA bound to heteroduplex substrates containing repeat loops of 1, 2, or 17 CAG repeats with a K(d) of about 10-20 nm, which is an affinity about 2 orders of magnitude higher than that of UvrA bound to the control substrates containing (CTG)(n).(CAG)(n) in the linear form. These results suggest that UvrA is involved in triplet repeat instability in cells. Specifically, UvrA may bind to loops formed during replication slippage or in slipped strand DNA and initiate DNA repair events that result in repeat deletion. These results imply a more comprehensive role for UvrA, in addition to the recognition of DNA damage, in maintaining the integrity of the genome. PMID- 11413148 TI - Transcription factor STAT5A is a substrate of Bruton's tyrosine kinase in B cells. AB - STAT5A is a molecular regulator of proliferation, differentiation, and apoptosis in lymphohematopoietic cells. Here we show that STAT5A can serve as a functional substrate of Bruton's tyrosine kinase (BTK). Purified recombinant BTK was capable of directly binding purified recombinant STAT5A with high affinity (K(d) = 44 nm), as determined by surface plasmon resonance using a BIAcore biosensor system. BTK was also capable of tyrosine-phosphorylating ectopically expressed recombinant STAT5A on Tyr(694) both in vitro and in vivo in a Janus kinase 3 independent fashion. BTK phosphorylated the Y665F, Y668F, and Y682F,Y683F mutants but not the Y694F mutant of STAT5A. STAT5A mutations in the Src homology 2 (SH2) and SH3 domains did not alter the BTK-mediated tyrosine phosphorylation. Recombinant BTK proteins with mutant pleckstrin homology, SH2, or SH3 domains were capable of phosphorylating STAT5A, whereas recombinant BTK proteins with SH1/kinase domain mutations were not. In pull-down experiments, only full-length BTK and its SH1/kinase domain (but not the pleckstrin homology, SH2, or SH3 domains) were capable of binding STAT5A. Ectopically expressed BTK kinase domain was capable of tyrosine-phosphorylating STAT5A both in vitro and in vivo. BTK mediated tyrosine phosphorylation of ectopically expressed wild type (but not Tyr(694) mutant) STAT5A enhanced its DNA binding activity. In BTK-competent chicken B cells, anti-IgM-stimulated tyrosine phosphorylation of STAT5 protein was prevented by pretreatment with the BTK inhibitor LFM-A13 but not by pretreatment with the JAK3 inhibitor HI-P131. B cell antigen receptor ligation resulted in enhanced tyrosine phosphorylation of STAT5 in BTK-deficient chicken B cells reconstituted with wild type human BTK but not in BTK-deficient chicken B cells reconstituted with kinase-inactive mutant BTK. Similarly, anti-IgM stimulation resulted in enhanced tyrosine phosphorylation of STAT5A in BTK competent B cells from wild type mice but not in BTK-deficient B cells from XID mice. In contrast to B cells from XID mice, B cells from JAK3 knockout mice showed a normal STAT5A phosphorylation response to anti-IgM stimulation. These findings provide unprecedented experimental evidence that BTK plays a nonredundant and pivotal role in B cell antigen receptor-mediated STAT5A activation in B cells. PMID- 11413149 TI - Stimulation of cytochrome P450 reactions by apo-cytochrome b5: evidence against transfer of heme from cytochrome P450 3A4 to apo-cytochrome b5 or heme oxygenase. AB - Many cytochrome P450 (P450)-dependent reactions have been shown to be stimulated by another microsomal protein, cytochrome b(5) (b(5)). Two major explanations are (i) direct electron transfer from b(5) and (ii) a conformational effect in the absence of electron transfer. Some P450s (e.g. 3A4, 2C9, 17A, and 4A7) are stimulated by either b(5) or b(5) devoid of heme (apo-b(5)), indicating a lack of electron transfer, whereas other P450s (e.g. 2E1) are stimulated by b(5) but not by apo-b(5). Recently, a proposal has been made by Guryev et al. (Biochemistry 40, 5018-5031, 2001) that the stimulation by apo-b(5) can be explained only by transfer of heme from P450 preparations to apo-b(5), enabling electron transfer. We have repeated earlier findings of stimulation of catalytic activity of testosterone 6beta-hydroxylation activities with four P450 preparations, in which nearly all of the heme was accounted for as P450. Spectral analysis of mixtures indicated that only approximately 5% of the heme can be transferred to apo-b(5), which cannot account for the observed stimulation. The presence of the heme scavenger apomyoglobin did not inhibit the stimulation of P450 3A4-dependent testosterone or nifedipine oxidation activity. Further evidence against the presence of loosely bound P450 3A4 heme was provided in experiments with apo-heme oxygenase, in which only 3% of the P450 heme was converted to biliverdin. Finally, b(5) supported NADH-b(5) reductase/P450 3A4-dependent testosterone 6beta hydroxylation, but apo-b(5) did not. Thus, apo-b(5) can stimulate P450 3A4 reactions as well as b(5) in the absence of electron transfer, and heme transfer from P450 3A4 to apo-b(5) cannot be used to explain the catalytic stimulation. PMID- 11413150 TI - PRMT5 (Janus kinase-binding protein 1) catalyzes the formation of symmetric dimethylarginine residues in proteins. AB - We have identified a new mammalian protein arginine N-methyltransferase, PRMT5, formerly designated Janus kinase-binding protein 1, that can catalyze the formation of omega-N(G)-monomethylarginine and symmetric omega-N(G),N(G') dimethylarginine in a variety of proteins. A hemagglutinin peptide-tagged PRMT5 complex purified from human HeLa cells catalyzes the S-adenosyl-l-[methyl (3)H]methionine-dependent in vitro methylation of myelin basic protein. When the radiolabeled myelin basic protein was acid-hydrolyzed to free amino acids, and the products were separated by high-resolution cation exchange chromatography, we were able to detect two tritiated species. One species co-migrated with a omega N(G)-monomethylarginine standard, and the other co-chromatographed with a symmetric omega-N(G),N(G')-dimethylarginine standard. Upon base treatment, this second species formed methylamine, a breakdown product characteristic of symmetric omega-N(G),N(G')-dimethylarginine. Further analysis of these two species by thin layer chromatography confirmed their identification as omega-N(G) monomethylarginine and symmetric omega-N(G),N(G')-dimethylarginine. The hemagglutinin-PRMT5 complex was also able to monomethylate and symmetrically dimethylate bovine histone H2A and a glutathione S-transferase-fibrillarin (amino acids 1-148) fusion protein (glutathione S-transferase-GAR). A mutation introduced into the S-adenosyl-l-methionine-binding motif I of a myc-tagged PRMT5 construct in COS-1 cells led to a near complete loss of observed enzymatic activity. PRMT5 is the first example of a catalytic chain for a type II protein arginine N-methyltransferase that can result in the formation of symmetric dimethylarginine residues as observed previously in myelin basic protein, Sm small nuclear ribonucleoproteins, and other polypeptides. PMID- 11413151 TI - Unorthodox routes to prostanoid formation: new twists in cyclooxygenase-initiated pathways. PMID- 11413152 TI - Why there are two cyclooxygenase isozymes. PMID- 11413153 TI - Acute graft-versus-host disease: inflammation run amok? PMID- 11413154 TI - Integrin-associated protein (CD47): an unusual activator of G protein signaling. PMID- 11413155 TI - Pulmonary fibrosis: a cellular overreaction or a failure of communication? PMID- 11413156 TI - ADP and platelets: the end of the beginning. PMID- 11413157 TI - TNF and receptors in organ-specific autoimmune disease: multi-layered functioning mirrored in animal models. PMID- 11413158 TI - FGF-8 in the ventral pharynx alters development of myocardial calcium transients after neural crest ablation. AB - Cardiac neural crest ablation results in depressed myocardial calcium transients and elevated proliferation in myocardium at a stage when cardiac neural crest cells are not in contact with the myocardium. To test the hypothesis that cardiac neural crest-derived cells, which migrate into the caudal, ventral pharynx at stage 14, block a signal from the ventral pharynx, we cultured stage 12 chick heart tube or myocardial strips in the presence or absence of ventral pharynx. We found that myocardium cultured with ventral pharynx that had not yet contacted neural crest cells had significantly reduced calcium transients and an increased rate of proliferation. Ventral pharynx from intact embryos at a stage when neural crest-derived cells had reached the pharynx had no effect on myocardial calcium transients. Ventral pharynx from neural crest-ablated embryos continued to suppress myocardial calcium transients at this later stage. Myocardium cultured with FGF-2 also showed a significant reduction in calcium transients. An FGF-2 neutralizing Ab reversed the deleterious effect of the ventral pharynx on myocardial calcium transients and proliferation. We therefore examined the expression of FGF-2 and similar FGFs in the ventral pharynx. Only FGF-8 was expressed in a temporospatial pattern that made it a viable candidate for altering the myocardial calcium transient during stages 14-18. In explant cultures, neutralizing Ab for FGF-8 rescued development of the myocardial calcium transient in neural crest-ablated chick embryos. PMID- 11413159 TI - Severe inflammatory arthritis and lymphadenopathy in the absence of TNF. AB - It has been postulated that TNF has a pivotal role in a cytokine cascade that results in joint inflammation and destruction in rheumatoid arthritis (RA). To evaluate this, we examined the response of TNF-deficient (Tnf(-/-)) mice in two models of RA. Collagen-induced arthritis (CIA) was induced by injection of chick type II collagen (CII) in CFA. Tnf(-/-) mice had some reduction in the clinical parameters of CIA and, on histology, significantly more normal joints. However, severe disease was evident in 54% of arthritic Tnf(-/-) joints. Tnf(-/-) mice had impaired Ig class switching, but preserved T cell proliferative responses to CII and enhanced IFN-gamma production. Interestingly, CII-immunized Tnf(-/-) mice developed lymphadenopathy and splenomegaly associated with increased memory CD4(+) T cells and activated lymph node B cells. Acute inflammatory arthritis was also reduced in Tnf(-/-) mice, although again some mice exhibited severe disease. We conclude that TNF is important but not essential for inflammatory arthritis; in each model, severe arthritis could proceed even in the complete absence of TNF. These results call into doubt the concept that TNF is obligatory for chronic autoimmune and acute inflammatory arthritis and provide a rationale for further studies into TNF-independent cytokine pathways in arthritis. PMID- 11413160 TI - Transient expression of IL-1beta induces acute lung injury and chronic repair leading to pulmonary fibrosis. AB - IL-1beta is one of a family of proinflammatory cytokines thought to be involved in many acute and chronic diseases. Although it is considered to participate in wound repair, no major role has been attributed to IL-1beta in tissue fibrosis. We used adenoviral gene transfer to transiently overexpress IL-1beta in rat lungs after intratracheal administration. The high expression of IL-1beta in the first week after injection was accompanied by local increase of the proinflammatory cytokines IL-6 and TNF-alpha and a vigorous acute inflammatory tissue response with evidence of tissue injury. The profibrotic cytokines PDGF and TGF-beta1 were increased in lung fluid samples 1 week after peak expression of IL-1beta. Although PDGF returned to baseline in the third week, TGF-beta1 showed increased concentrations in bronchoalveolar lavage fluid for up to 60 days. This was associated with severe progressive tissue fibrosis in the lung, as shown by the presence of myofibroblasts, fibroblast foci, and significant extracellular accumulations of collagen and fibronectin. These data directly demonstrate how acute tissue injury in the lung, initiated by a highly proinflammatory cytokine, IL-1beta, converts to progressive fibrotic changes. IL-1beta should be considered a valid target for therapeutic intervention in diseases associated with fibrosis and tissue remodeling. PMID- 11413161 TI - TGF-beta is a critical mediator of acute lung injury. AB - We have shown that the integrin alphavbeta6 activates latent TGF-beta in the lungs and skin. We show here that mice lacking this integrin are completely protected from pulmonary edema in a model of bleomycin-induced acute lung injury (ALI). Pharmacologic inhibition of TGF-beta also protected wild-type mice from pulmonary edema induced by bleomycin or Escherichia coli endotoxin. TGF-beta directly increased alveolar epithelial permeability in vitro by a mechanism that involved depletion of intracellular glutathione. These data suggest that integrin mediated local activation of TGF-beta is critical to the development of pulmonary edema in ALI and that blocking TGF-beta or its activation could be effective treatments for this currently untreatable disorder. PMID- 11413162 TI - Iron overload diminishes atherosclerosis in apoE-deficient mice. AB - It has been proposed that elevated levels of tissue iron increase the risk for atherosclerosis, perhaps by favoring the formation of pro-atherogenic oxidized LDL. Working with apoE-deficient (apoE(-/-)) mice, which do not require a high fat diet to develop atherosclerosis, we compared the effects of standard diet (0.02% iron) or a 2% carbonyl iron diet. After 24 weeks, mice fed the 2% carbonyl iron diet had twice as much iron in their plasma, a ninefold increase in bleomycin-detectable free iron in their plasma, and ten times as much iron in their livers as control mice. Dietary iron overload caused a modest (30%) rise in plasma triglyceride and cholesterol. Nevertheless, this regimen did not exacerbate, but rather reduced the severity of atherosclerosis by 50%, and it failed to elevate hepatic levels of heme oxygenase mRNA, which is induced by many different oxidative insults in vitro. Moreover, hepatic levels of protein-bound dityrosine and ortho-tyrosine, two markers of metal-catalyzed oxidative damage in vitro, failed to rise in iron-overloaded animals. Our observations suggest that elevated serum and tissue levels of iron are not atherogenic in apoE(-/-) mice. Moreover, they call into question the hypothesis that elevated levels of tissue iron promote LDL oxidation and oxidative stress in vivo. PMID- 11413163 TI - Activation of sickle red blood cell adhesion via integrin-associated protein/CD47 induced signal transduction. AB - Peripheral human red blood cells (RBCs) are not generally known to become activated and adhesive in response to cell signaling. We show, however, that soluble thrombospondin via integrin-associated protein (IAP; CD47) increases the adhesiveness of sickle RBCs (SS RBCs) by activating signal transduction in the SS RBC. This stimulated adhesion requires occupancy of IAP and shear stress and is mediated by the activation of large G proteins and tyrosine kinases. Reticulocyte enriched RBCs derived from sickle-cell disease (SCD) patients are most responsive to IAP-induced activation. These studies therefore establish peripheral SS RBCs as signaling cells that respond to a novel synergy between IAP-induced signal transduction and shear stress, suggesting new therapeutic targets in SCD. PMID- 11413164 TI - Endothelin-1/endothelin-B receptor-mediated increases in NHE3 activity in chronic metabolic acidosis. AB - Decreases in blood pH activate NHE3, the proximal tubular apical membrane Na/H antiporter. In cultured renal epithelial cells, activation of the endothelin-B (ET(B)) receptor increases NHE3 activity. To examine the role of the ET(B) receptor in the response to acidosis in vivo, the present studies examined ET(B) receptor-deficient mice, rescued from neonatal lethality by expression of a dopamine beta-hydroxylase promoter/ET(B) receptor transgene (Tg/Tg:ET(B)(-/-) mice). In proximal tubule suspensions from Tg/Tg:ET(B)(+/-) mice, 10(-8) M endothelin-1 (ET-1) increased NHE3 activity, but this treatment had no effect on tubules from Tg/Tg:ET(B)(-/-) mice. Acid ingestion for 7 days caused a greater decrease in blood HCO(3)(-) concentration in Tg/Tg:ET(B)(-/-) mice compared with Tg/Tg:ET(B)(+/+) and Tg/Tg:ET(B)(+/-) mice. Whereas acid ingestion increased apical membrane NHE3 by 42-46% in Tg/Tg:ET(B)(+/+) and Tg/Tg:ET(B)(+/-) mice, it had no effect on NHE3 in Tg/Tg:ET(B)(-/-) mice. In C57BL/6 mice, excess acid ingestion increased renal cortical preproET-1 mRNA expression 2.4-fold and decreased preproET-3 mRNA expression by 37%. On a control diet, Tg/Tg:ET(B)(-/-) mice had low rates of ammonium excretion, which could not be attributed to an inability to acidify the urine, as well as hypercitraturia, with increased titratable acid excretion. Acid ingestion increased ammonium excretion, citrate absorption, and titratable acid excretion to the same levels in Tg/Tg:ET(B)(-/-) and Tg/Tg:ET(B)(+/+) mice. In conclusion, metabolic acidosis increases ET-1 expression, which increases NHE3 activity via the ET(B) receptor. PMID- 11413165 TI - Somatostatin is required for masculinization of growth hormone-regulated hepatic gene expression but not of somatic growth. AB - Pulsatile growth hormone (GH) secretion differs between males and females and regulates the sex-specific expression of cytochrome P450s in liver. Sex steroids influence the secretory dynamics of GH, but the neuroendocrine mechanisms have not been conclusively established. Because periventricular hypothalamic somatostatin (SST) expression is greater in males than in females, we generated knockout (Smst(-/-)) mice to investigate whether SST peptides are necessary for sexually differentiated GH secretion and action. Despite marked increases in nadir and median plasma GH levels in both sexes of Smst(-/-) compared with Smst(+/+) mice, the mutant mice had growth curves identical to their sibling controls and retained a normal sexual dimorphism in weight and length. In contrast, the liver of male Smst(-/-) mice was feminized, resulting in an identical profile of GH-regulated hepatic mRNAs between male and female mutants. Male Smst(-/-) mice show higher expression of two SST receptors in the hypothalamus and pituitary than do females. These data indicate that SST is required to masculinize the ultradian GH rhythm by suppressing interpulse GH levels. In the absence of SST, male and female mice exhibit similarly altered plasma GH profiles that eliminate sexually dimorphic liver function but do not affect dimorphic growth. PMID- 11413166 TI - LPS antagonism reduces graft-versus-host disease and preserves graft-versus leukemia activity after experimental bone marrow transplantation. AB - Acute graft-versus-host disease (GVHD) and leukemic relapse remain the two major obstacles to successful outcomes after allogeneic bone marrow transplantation (BMT). Recent studies have demonstrated that the loss of gastrointestinal tract integrity, and specifically the translocation of LPS into the systemic circulation, is critical to the induction of cytokine dysregulation that contributes to GVHD. Using a mouse BMT model, we studied the effects of direct LPS antagonism on GVHD severity and graft-versus-leukemia (GVL) activity. Administration of B975, a synthetic lipid-A analogue from day 0 to day +6, reduced serum TNF-alpha levels, decreased intestinal histopathology, and resulted in significantly improved survival and a reduction in clinical GVHD, compared with control-treated animals. Importantly, B975 had no effect on donor T cell responses to host antigens in vivo or in vitro. When mice received lethal doses of P815 tumor cells at the time of BMT, administration of B975 did not impair GVL activity and resulted in significantly improved leukemia-free survival. These findings reveal a critical role for LPS in the early inflammatory events contributing to GVHD and suggest that a new class of pharmacologic agents, LPS antagonists, may help to prevent GVHD while preserving T cell responses to host antigens and GVL activity. PMID- 11413167 TI - Molecular identification and characterization of the platelet ADP receptor targeted by thienopyridine antithrombotic drugs. AB - ADP plays a critical role in modulating thrombosis and hemostasis. ADP initiates platelet aggregation by simultaneous activation of two G protein-coupled receptors, P2Y1 and P2Y12. Activation of P2Y1 activates phospholipase C and triggers shape change, while P2Y12 couples to Gi to reduce adenylyl cyclase activity. P2Y12 has been shown to be the target of the thienopyridine drugs, ticlopidine and clopidogrel. Recently, we cloned a human orphan receptor, SP1999, highly expressed in brain and platelets, which responded to ADP and had a pharmacological profile similar to that of P2Y12. To determine whether SP1999 is P2Y12, we generated SP1999-null mice. These mice appear normal, but they exhibit highly prolonged bleeding times, and their platelets aggregate poorly in responses to ADP and display a reduced sensitivity to thrombin and collagen. These platelets retain normal shape change and calcium flux in response to ADP but fail to inhibit adenylyl cyclase. In addition, oral clopidogrel does not inhibit aggregation responses to ADP in these mice. These results demonstrate that SP1999 is indeed the elusive receptor, P2Y12. Identification of the target receptor of the thienopyridine drugs affords us a better understanding of platelet function and provides tools that may lead to the discovery of more effective antithrombotic therapies. PMID- 11413168 TI - A viral protein that selectively downregulates ICAM-1 and B7-2 and modulates T cell costimulation. AB - Kaposi's sarcoma-associated (KS-associated) herpesvirus (KSHV) is a B lymphotropic agent linked to AIDS-related lymphoproliferative disorders and KS. We and others have earlier identified two viral genes, K3 and K5, that encode endoplasmic reticulum proteins that downregulate surface MHC-I chains by enhancing their endocytosis. Here we have examined the ability of these proteins to influence the disposition of other host surface proteins implicated in immune recognition and activation. We report that K5, but not K3, expression in BJAB cells dramatically reduces ICAM-1 and B7-2 surface expression; B7-1 expression is unaffected. This K5-induced reduction can be reversed by coexpression of a dominant negative mutant of dynamin, indicating that the loss of ICAM and B7-2 surface expression is due to their enhanced endocytosis. This downregulation is functionally significant, because K5-transfected B cells show substantial impairment in their ability to induce T cell activation. K5 is thus the first example of a viral modulator of immunological synapse formation and T cell costimulation. We propose that its expression reduces T cell responses to KSHV infected B cells early in infection, thereby diminishing antiviral cytokine release and the production of stimulatory signals for CTL generation. PMID- 11413171 TI - Addressing health inequalities in health impact assessment. PMID- 11413173 TI - Basic concepts in population health and health care. PMID- 11413174 TI - Formal education and back pain: a review. AB - OBJECTIVES: To summarise the scientific evidence on the relation between educational status and measures of the frequency and the consequences of back pain and of the outcomes of interventions among back pain patients, and to outline possible mechanisms that could explain such an association if found. DESIGN: Sixty four articles published between 1966 and 2000 that documented the association of formal education with back pain were reviewed. MAIN RESULTS: Overall, the current available evidence points indirectly to a stronger association of low education with longer duration and/or higher recurrence of back pain than to an association with onset. The many reports of an association of low education with adverse consequences of back pain also suggest that the course of a back pain episode is less favourable among persons with low educational attainment. Mechanisms that could explain these associations include variations in behavioural and environmental risk factors by educational status, differences in occupational factors, compromised "health stock" among people with low education, differences in access to and utilisation of health services, and adaptation to stress. Although lower education was not associated with the outcomes of interventions in major studies, it is difficult, in light of the current limited available evidence, to draw firm conclusions on this association. CONCLUSION: Scientific evidence supports the hypothesis that less well educated people are more likely to be affected by disabling back pain. Further study of this association may help advance our understanding of back pain as well as understanding of the relation between socioeconomic status and disease as a general phenomenon. PMID- 11413175 TI - The incidence of childhood leukaemia around the La Hague nuclear waste reprocessing plant (France): a survey for the years 1978-1998. AB - BACKGROUND: A previous study has suggested an increased incidence rate of leukaemia from 1978 to 1992 in people aged 0 to 24 years and living in the vicinity of the La Hague nuclear waste reprocessing plant without considering age and cytological type. SETTING: The Nord Cotentin region (France) and the island of Alderney (United Kingdom). STUDY OBJECTIVE: To describe the occurrence of leukaemia for each age group and cytological type from 1978 to 1998 in the same area, using accurate reference incidence rates and adequate estimation of the at risk population. DESIGN: A geographical study of incidence using three zones defined according to their distance from the site (0 to 10 km: Beaumont-Hague electoral ward, 10 to 20 km and 20 to 35 km) has been conducted. The risk of leukaemia was estimated from the standardised incidence ratio (SIR) of the number of cases observed to the number expected. Exact 95% confidence intervals (CI) have been computed. PARTICIPANTS: All people under the age of 25 years living in the study region between 1978 and 1998. MAIN RESULTS: The observed number of cases of leukaemia in the study region as a whole was consistent with the expected value (SIR=1.03; 95%CI: 0.73, 1.41). No cases were observed on Alderney. The SIR in the Beaumont-Hague electoral ward was 2.17 (95%CI: 0.71, 5.07). The highest SIR was observed in the 5 to 9 years age group (SIR=6.38; 95%CI: 1.32, 18.65). This consists in acute lymphoblastic leukaemia cases. CONCLUSION: This study indicates an increased incidence of leukaemia in the area situated at less than 10 km from the plant. Monitoring and further investigations should be targeted at acute lymphoblastic leukaemia occurring during the childhood incidence peak (before 10 years) in children living near the La Hague site and may be other nuclear reprocessing plants. PMID- 11413176 TI - Relation of socioeconomic position to the case fatality, prognosis and treatment of myocardial infarction events; the FINMONICA MI Register Study. AB - OBJECTIVE: To examine socioeconomic differences in case fatality and prognosis of myocardial infarction (MI) events, and to estimate the contributions of incidence and case fatality to socioeconomic differences in coronary heart disease (CHD) mortality. DESIGN: A population-based MI register study. METHODS: The FINMONICA MI Register recorded all MI events among persons aged 35-64 years in three areas of Finland during 1983-1992. A record linkage of the MI Register data with the files of Statistics Finland was performed to obtain information on socioeconomic indicators for each individual registered. First MI events (n=8427) were included in the analyses. MAIN RESULTS: The adjusted risk ratio of prehospital coronary death was 2.11 (95% CI 1.82, 2.46) among men and 1.68 (1.14, 2.48) among women with low income compared with those with high income. Even among persons hospitalised alive the risk of death during the next 12 months was markedly higher in the low income group than in the high income group. Case fatality explained 51% of the CHD mortality difference between the low and the high income groups among men and 38% among women. Incidence contributed 49% and 62%, respectively. CONCLUSIONS: Considerable socioeconomic differences were observed in the case fatality of first coronary events both before hospitalisation and among patients hospitalised alive. Case fatality explained a half of the CHD mortality difference between the low and the high income groups among men and more than a third among women. PMID- 11413177 TI - Trends in the hospital management of unstable angina. AB - STUDY OBJECTIVE: To examine the hospital management of unstable angina (UAP) in 1996 and 1998, according to patient demographic variables and disease severity. DESIGN: Medical record review. SETTING: 37 hospitals across New South Wales, Australia, representative of the secondary and tertiary care hospitals in the State. PARTICIPANTS: All patients (or a random sample of patients) with UAP admitted to these hospitals during five months in 1996 and six months in 1998 (1872 and 1368 patients respectively). MAIN RESULTS: In the two years between 1996 and 1998, there was an increase in the use of beta blockers and a corresponding decrease in the use of calcium channel blockers, as well as a decrease in the use of intravenous nitrates. Those aged 75 or more were roughly half as likely as those aged less than 65 to be prescribed heparin, aspirin and heparin, beta blockers, intravenous nitrates, and only one third as likely to be offered coronary angiography in hospital. They were one and a half times as likely to be prescribed calcium channel blockers compared with the youngest age group. A similar pattern was seen for gender, where men were more likely than women to be given aspirin, aspirin and heparin, and coronary angiography, and less likely to be given calcium channel blockers. Those with a past history were less likely, and those with more severe disease were more likely than others to be given most interventions. CONCLUSIONS: In view of the low use of evidence based management of UAP among women and the elderly, it would seem appropriate for disease management guidelines to target these groups. PMID- 11413178 TI - Role of age and sex in short-term and long term mortality after a first Q wave myocardial infarction. AB - STUDY OBJECTIVE: The objective of this study was to analyse whether the risk of death within 28 days and three years after a first Q wave myocardial infarction was higher in hospitalised women than in men. DESIGN: Follow up study. PATIENTS AND SETTING: All consecutive first Q wave myocardial infarction patients aged 25 to 74 years (447 women and 2322 men) admitted to a tertiary hospital in Gerona, Spain, from 1978 to 1997 were registered and followed up for three years. MAIN RESULTS: Women were older, presented more comorbidity and developed more severe myocardial infarctions than men. A significant interaction was found between sex and age. Women aged 65-74 had higher early mortality risk than men of the same age (OR 1.62; 95% CI 1.01, 2.66) after adjusting for age, comorbidity and acute complications including heart failure. Women under 65 tended to be at lower risk of early mortality than men (0.45 (95% CI 0.19, 1.04). Three year mortality of 28 day survivors did not differ between sexes. CONCLUSIONS: These data support the idea that the higher 28 day mortality in hospitalised women with a first Q wave myocardial infarction is mainly attributable to the large number of patients aged 65 to 74 years in whom the risk is higher than that in men. Women under 65 with myocardial infarction do not seem to be a special group of risk. PMID- 11413179 TI - Change in male and female life expectancy by social class: decomposition by age and cause of death in Finland 1971-95. AB - STUDY OBJECTIVE: To quantify the contribution of different causes of death and age groups for trends in life expectancy for two major social classes. DESIGN AND SETTING: Prospective study of mortality in Finland among all over 35 year old men and women. Baseline social class (manual/non-manual) was from the 1970, 1975, 1980, 1985 and 1990 census records, and follow up was by computerised record linkage to death certificates for 1971-1995. MAIN RESULTS: From the early 1970s to the early 1990s life expectancy at age 35 increased by about five and four years among Finnish men and women respectively, with largest gains among 55-74 year old men and 65-84 year old women. Life expectancy increase was 5.1 years among non-manual and 3.8 years among manual men; corresponding figures for women were 3.6 and 3.0 years. In the 1980s, when differences in life expectancy increased most rapidly, decline in cardiovascular disease mortality was more rapid in the non-manual than the manual class. Furthermore, increasing mortality for alcohol associated causes, "other diseases", and accidents and violence were most prominent in the manual class. CONCLUSIONS: Explanations of increasing social inequalities in mortality that are based on one underlying factor are difficult to reconcile with the variability in the cause specific trends in social inequalities in mortality. The contribution of older ages to social inequalities in mortality should be more widely recognised. PMID- 11413180 TI - Results from a family and DNA based active identification programme for familial hypercholesterolaemia. PMID- 11413181 TI - Individual employment histories and subsequent cause specific hospital admissions and mortality: a prospective study of a cohort of male and female workers with 21 years follow up. PMID- 11413182 TI - Height and health: predicting longevity from bone length in archaeological remains. PMID- 11413183 TI - A heuristic approach to the formulas for population attributable fraction. AB - BACKGROUND: As the definitional formula for population attributable fraction is not usually directly usable in applications, separate estimation formulas are required. However, most epidemiology textbooks limit their coverage to Levin's formula, based on the (dichotomous) distribution of the exposure of interest in the population. Few present or explain Miettinen's formula, based on the distribution of the exposure in the cases; and even fewer present the corresponding formulas for situations with more than two levels of exposure. Thus, many health researchers and public health practitioners are unaware of, or are not confident in their use of, these formulas, particularly when they involve several exposure levels, or confounding factors. METHODS/RESULTS: A heuristic approach, coupled with pictorial representations, is offered to help understand and interconnect the structures behind the Levin and Miettinen formulas. The pictorial representation shows how to deal correctly with several exposure levels, and why a commonly used approach is incorrect. Correct and incorrect approaches are also presented for situations where estimates must be aggregated over strata of a confounding factor. PMID- 11413184 TI - The co-occurrence of AIDS and homelessness: results from the integration of administrative databases for AIDS surveillance and public shelter utilisation in Philadelphia. AB - STUDY OBJECTIVE: Administrative databases from the City of Philadelphia that track public shelter utilisation (n=44 337) and AIDS case reporting (n=7749) were merged to identify rates and risk factors for co-occurring homelessness and AIDS. DESIGN: Multiple decrement life tables analyses were conducted, and logistic regression analyses used to identify risk factors associated with AIDS among the homeless, and homelessness among people with AIDS. SETTING: City of Philadelphia, Pennsylvania, USA. MAIN RESULTS: People admitted to public shelters had a three year rate of subsequent AIDS diagnosis of 1.8 per 100 person years; nine times the rate for the general population of Philadelphia. Logistic regression results show that substance abuse history (OR = 3.14), male gender (OR = 2.05), and a history of serious mental disorder (OR = 1.62) were significantly related to the risk for AIDS diagnosis among shelter users. Among people with AIDS, results show a three year rate of subsequent shelter admission of 6.9 per 100 person years, and a three year rate of prior shelter admission of 9%, three times the three year rate of shelter admission for the general population. Logistic regression results show that intravenous drug user history (OR = 3.14); no private insurance (OR = 2.93); black race (OR = 2.82); pulmonary or extra-pulmonary TB (OR = 1.43); and pneumocystis pneumonia (OR = 0.56) were all related to the risk for shelter admission. CONCLUSIONS: Homelessness prevention programmes should target people with HIV risk factors, and HIV prevention programmes should be targeted to homeless persons, as these populations have significant intersection. Reasons and implications for this intersection are discussed. PMID- 11413185 TI - Is it feasible to plan secondary care services for coronary heart disease rationally? A quantified modelling approach for a UK Health Authority. AB - BACKGROUND: Coronary heart disease (CHD) is the major cause of mortality in the UK. This paper explores the difficulties facing health authorities in applying a rational and needs based approach to the planning of hospital based services and describes a simple model used to bring available information to bear on this problem. METHOD: Published estimates of CHD incidence were identified and methodologies were critically appraised. Estimates were extrapolated to a district population. A three month cohort study of patients with suspected CHD was undertaken within a district general hospital and a model of these clinical pathways was used to examine the volumes of patients and services required to meet the estimated levels of need. RESULTS: From published studies, estimates of CHD incidence ranged from 83 to 3600 per 100 000. From the cohort study, of patients referred with possible CHD 62% received a definitive diagnosis of CHD, 56% underwent an exercise ECG, 16% received an angiogram, 4% received a CABG and 2% a PTCA. Using these figures together with the cohort study, estimated activity ranges from 247 to 6475 surgical interventions per million population compared with the National Service Framework for Coronary Heart Disease recommendations of 1500 procedures per million. CONCLUSIONS: Current research on CHD incidence gives a very wide variation in estimated need. This makes its value for service planning questionable and the model highlights a need for further high quality research. The model provides a link between epidemiological research and secondary care service planning and supports the implementation of recommendations within the National Service Framework for Coronary Heart Disease. PMID- 11413189 TI - Control of von Willebrand factor multimer size by thrombospondin-1. AB - Plasma von Willebrand factor (vWF) is a multimeric protein that mediates adhesion of platelets to sites of vascular injury. Only the very large vWF multimers are effective in promoting platelet adhesion in flowing blood. A protein disulfide bond reductase in plasma reduces the average multimer size of vWF secreted by endothelial cells. This activity has been isolated from human endothelial cell conditioned medium and shown to be the trimeric glycoprotein, thrombospondin-1 (TSP-1). Incubation of purified TSP-1 with vWF resulted in formation of thiol dependent complexes of TSP-1 and vWF, generation of new thiols in vWF, and reduction in the average multimer size of vWF. The ratio of the concentrations of TSP-1 and vWF in plasma reflected with average multimer size of vWF. The higher the plasma TSP-1/vWF molar ratio, the smaller the average vWF multimer size. In addition, administration of TSP-1 to mice resulted in reduction in the average multimer size of plasma vWF. Interaction of TSP-1 with vWF is mediated by TSP-1 type 1 properdin domains and the vWF A3 domain. These results indicate that TSP-1 regulates the multimeric size and therefore hemostatic activity of vWF. PMID- 11413190 TI - Two waves of nuclear factor kappaB recruitment to target promoters. AB - Proinflammatory stimuli induce the rapid and transient translocation of nuclear factor (NF)-kappaB to the nucleus, where it activates transcription from several genes, including those encoding inflammatory cytokines and chemokines, adhesion molecules, and cytoprotective proteins. Using chromatin immunoprecipitation, we show that after an acute stimulation two distinct waves of NF-kappaB recruitment to target promoters occur: a fast recruitment to constitutively and immediately accessible (CIA) promoters and a late recruitment to promoters requiring stimulus dependent modifications in chromatin structure to make NF-kappaB sites accessible (promoters with regulated and late accessibility [RLA]). Our results suggest that a mechanism of specificity in NF-kappaB-dependent transcriptional responses relies on the ability of individual stimuli to make RLA promoters accessible to NF-kappaB before its rapid extrusion from the nucleus. PMID- 11413191 TI - Role of promyelocytic leukemia (PML) sumolation in nuclear body formation, 11S proteasome recruitment, and As2O3-induced PML or PML/retinoic acid receptor alpha degradation. AB - Promyelocytic leukemia (PML) is the organizer of nuclear matrix domains, PML nuclear bodies (NBs), with a proposed role in apoptosis control. In acute promyelocytic leukemia, PML/retinoic acid receptor (RAR) alpha expression disrupts NBs, but therapies such as retinoic acid or arsenic trioxide (As2O3) restore them. PML is conjugated by the ubiquitin-related peptide SUMO-1, a process enhanced by As2O3 and proposed to target PML to the nuclear matrix. We demonstrate that As2O3 triggers the proteasome-dependent degradation of PML and PML/RARalpha and that this process requires a specific sumolation site in PML, K160. PML sumolation is dispensable for its As2O3-induced matrix targeting and formation of primary nuclear aggregates, but is required for the formation of secondary shell-like NBs. Interestingly, only these mature NBs harbor 11S proteasome components, which are further recruited upon As2O3 exposure. Proteasome recruitment by sumolated PML only likely accounts for the failure of PML-K160R to be degraded. Therefore, studying the basis of As2O3-induced PML/RARalpha degradation we show that PML sumolation directly or indirectly promotes its catabolism, suggesting that mature NBs could be sites of intranuclear proteolysis and opening new insights into NB alterations found in viral infections or transformation. PMID- 11413192 TI - Subspecialization of CXCR5+ T cells: B helper activity is focused in a germinal center-localized subset of CXCR5+ T cells. AB - The T helper (Th) cell pool is composed of specialized cells with heterogeneous effector functions. Apart from Th1 and 2 cells, CXCR5+ T cells have been suggested to be another type of effector T cell specialized for B cell help. We show here that CXCR5+ T cells are heterogeneous, and we identify subsets of CXCR5+ CD4 T cells that differ in function and microenvironmental localization in secondary lymphoid tissues. CD57+CXCR5 T cells, hereafter termed germinal center Th (GC-Th) cells, are localized only in GCs, lack CCR7, and are highly responsive to the follicular chemokine B lymphocyte chemoattractant but not to the T cell zone EBI1-ligand chemokine. Importantly, GC-Th cells are much more efficient than CD57-CXCR5+ T cells or CXCR5- T cells in inducing antibody production from B cells. Consistent with their function, GC-Th cells produce elevated levels of interleukin 10 upon stimulation which, with other cytokines and costimulatory molecules, may help confer their B cell helper activity. Our results demonstrate that CXCR5+ T cells are functionally heterogeneous and that the GC-Th cells, a small subset of CXCR5+ T cells, are the key helpers for B cell differentiation and antibody production in lymphoid tissues. PMID- 11413193 TI - c-abl is required for the development of hyperoxia-induced retinopathy. AB - The requirement for the nonreceptor tyrosine kinase c-abl in the pathogenesis of retinopathy of prematurity (ROP) was examined using the mouse model for ROP and c abl-deficient mice. Hyperoxia-induced retinal neovascularization was observed in wild-type and heterozygous mice but animals that were homozygous null for c-abl did not develop a vasoproliferative retinopathy in response to hyperoxia. Two gene products, endothelin-1 (ET-1) and vascular endothelial growth factor (VEGF), have been implicated in the pathogenesis of ROP. The mRNA expression of ET-1 and VEGF was assessed in mice maintained in normoxia and in hyperoxia-exposed mice. ET-1 mRNA levels were unchanged in wild-type mice throughout the hyperoxia treatment, suggesting that ET-1 mRNA expression is not regulated by the increase in inspired oxygen. In wild-type mice maintained in room air, VEGF mRNA levels rose threefold from postnatal day 6 (P6) to P17. When wild-type mice were treated with the hyperoxia regimen, a fivefold decrease in VEGF mRNA expression was observed from P7 to P16. However, retinal VEGF expression in hyperoxia-treated homozygous null mice did not decrease and remained at control levels. These data suggest that c-abl is required for the hyperoxia-induced retinal neovascularization and hyperoxia-induced decrease in VEGF mRNA levels. PMID- 11413194 TI - Aberrant high expression of B lymphocyte chemokine (BLC/CXCL13) by C11b+CD11c+ dendritic cells in murine lupus and preferential chemotaxis of B1 cells towards BLC. AB - We observed here that the expression of B lymphocyte chemokine (BLC/CXCL13) was markedly enhanced in the thymus and kidney in aged (NZB x NZW)F1 (BWF1) mice developing lupus nephritis, but not in similarly aged NZB and NZW mice. BLC positive cells were present in the cellular infiltrates in the target organs with a reticular pattern of staining. CD11b+CD11c+ dendritic cells were increased in the thymus and spleen in aged BWF1 mice and identified as the major cell source for BLC. CD4+ T cells as well as B cells were dramatically increased in the thymus in aged BWF1 mice, whereas no increase was observed in aged NZB and NZW mice. B1/B2 ratio in the thymus was significantly higher than those in the spleen and peripheral blood in aged BWF1 mice. Interestingly, BLC showed preferential chemotactic activity for B1 cells derived from several mouse strains, including nonautoimmune mice. Cell surface CXCR5 expression on B1 cells was significantly higher than that on B2 cells. Thus, aberrant high expression of BLC by myeloid dendritic cells in the target organs in aged BWF1 mice may play a pivotal role in breaking immune tolerance in the thymus and in recruiting autoantibody-producing B cells in the development of murine lupus. PMID- 11413195 TI - Antibodies against merozoite surface protein (MSP)-1(19) are a major component of the invasion-inhibitory response in individuals immune to malaria. AB - Antibodies that bind to antigens expressed on the merozoite form of the malaria parasite can inhibit parasite growth by preventing merozoite invasion of red blood cells. Inhibitory antibodies are found in the sera of malaria-immune individuals, however, the specificity of those that are important to this process is not known. In this paper, we have used allelic replacement to construct a Plasmodium falciparum parasite line that expresses the complete COOH-terminal fragment of merozoite surface protein (MSP)-1(19) from the divergent rodent malaria P. chabaudi. By comparing this transfected line with parental parasites that differ only in MSP-1(19), we show that antibodies specific for this domain are a major component of the inhibitory response in P. falciparum-immune humans and P. chabaudi-immune mice. In some individual human sera, MSP-1(19) antibodies dominated the inhibitory activity. The finding that antibodies to a small region of a single protein play a major role in this process has important implications for malaria immunity and is strongly supportive of further understanding and development of MSP-1(19)-based vaccines. PMID- 11413196 TI - Functional dichotomy in natural killer cell signaling: Vav1-dependent and independent mechanisms. AB - The product of the protooncogene Vav1 participates in multiple signaling pathways and is a critical regulator of antigen-receptor signaling in B and T lymphocytes, but its role during in vivo natural killer (NK) cell differentiation is not known. Here we have studied NK cell development in Vav1-/- mice and found that, in contrast to T and NK-T cells, the absolute numbers of phenotypically mature NK cells were not reduced. Vav1-/- mice produced normal amounts of interferon (IFN) gamma in response to Listeria monocytogenes and controlled early infection but showed reduced tumor clearance in vivo. In vitro stimulation of surface receptors in Vav1-/- NK cells resulted in normal IFN-gamma production but reduced tumor cell lysis. Vav1 was found to control activation of extracellular signal regulated kinases and exocytosis of cytotoxic granules. In contrast, conjugate formation appeared to be only mildly affected, and calcium mobilization was normal in Vav1-/- NK cells. These results highlight fundamental differences between proximal signaling events in T and NK cells and suggest a functional dichotomy for Vav1 in NK cells: a role in cytotoxicity but not for IFN-gamma production. PMID- 11413197 TI - A transcription function for the T cell-specific adapter (TSAd) protein in T cells: critical role of the TSAd Src homology 2 domain. AB - T cell-specific adapter (TSAd) protein is an Src homology 2 (SH2) domain containing adapter molecule implicated in T cell receptor for antigen (TCR) mediated interleukin 2 (IL-2) secretion in T cells. Here, we demonstrate that a substantial fraction of TSAd is found in the T cell nucleus. Nuclear import of TSAd is an active process that depends on TSAd SH2 domain recognition of a phosphotyrosine-containing ligand. Importantly, we show that TSAd can act as a potent transcriptional activator in T cells. Furthermore, the TSAd SH2 domain appears to be essential for this transcription-activating function independent of its role in nuclear import. Biochemical analyses suggest that a single TSAd SH2 domain ligand of 95-100 kD may be involved in these processes. Consistent with a role as a transcription activator, cotransfection of TSAd with an IL-2 promoter reporter gene construct results in a considerable upregulation of IL-2 promoter activity. Further, we show that this augmentation requires a functional TSAd SH2 domain. However, TSAd does not appear to modulate the activity of the major recognized IL-2 gene transcription factors, nuclear factor kappaB (NF-kappaB), nuclear factor of activated T cells (NFAT), or activator protein 1 (AP-1). These findings point to the function of TSAd as a novel transcription-regulatory protein in T cells and illustrate the importance of the TSAd SH2 domain in this role. PMID- 11413198 TI - Essential requirement for c-kit and common gamma chain in thymocyte development cannot be overruled by enforced expression of Bcl-2. AB - The thymus in mice lacking both the receptor tyrosine kinase c-kit and the common cytokine receptor gamma chain (gamma(c)) is alymphoid because these receptors provide essential signals at the earliest stages of thymocyte development. The signals transduced by these receptors potentially regulate proliferation, survival, or differentiation, but the contribution of each receptor to distinct intracellular signaling cascades is only poorly defined. Here, we have examined whether enforced expression of Bcl-2 can rescue thymocyte development in c-kit and gamma(c) single or double mutant mice. A bcl-2 transgene (E(mu)-bcl-2-25; expressed in the T cell lineage) was introduced into (a) c-kit and gamma(c) wild type (c-kit+gamma(c)+bcl+), (b) c-kit-deficient (c-kit(-)gamma(c)+bcl+), (c) gamma(c)-deficient (c-kit+gamma(c)-bcl+), or (d) c-kit and gamma(c) double deficient mice (c-kit-gamma(c)-bcl+). The bcl-2 transgene was functionally active in wild-type and c-kit or gamma(c) single mutants, as it promoted survival of ex vivo isolated thymocytes, including pro-T cells. In vivo, however, transgenic Bcl 2 did not release T cell precursors from their phenotypic block and failed to increase progenitor or total thymocyte cellularity in c-kit or gamma(c) single or double mutants. These data argue strongly against a role for Bcl-2 as a key mediator in signaling pathways linked to cytokine and growth factor receptors driving early thymocyte development. PMID- 11413199 TI - On the role of the innate immunity in autoimmune disease. PMID- 11413200 TI - A robust neutralization test for Plasmodium falciparum malaria. PMID- 11413201 TI - Improving quantitative flowering models through a better understanding of the phases of photoperiod sensitivity. AB - A quantitative understanding of the phases of sensitivity to photo-thermal environment is important if the accuracy of flowering models is to be improved and if the timing of long and short day treatments in protected cropping is to be optimized. A simple method of quantifying the duration of the phases of sensitivity to photoperiod is through the use of reciprocal transfer experiments where plants are transferred between long and short days at regular intervals throughout development. The advantages and disadvantages of different analytical approaches used to analyse such data sets are examined. Inconsistencies between the approaches are highlighted, as are differences in the way authors have interpreted data. The problem of confounding the effects of photoperiod and light integral is considered, as is the need to separate the number of inductive cycles needed for flower commitment from the length of the juvenile phase. The effects of photo-thermal environment on the duration of these phases of photoperiod sensitivity are discussed, together with topics requiring further development. PMID- 11413202 TI - Down-regulation of a ripening-related beta-galactosidase gene (TBG1) in transgenic tomato fruits. AB - Exo-galactanase/beta-galactosidase (EC 3.2.1.23) activity is thought to be responsible for the loss of galactosyl residues from the cell walls of ripening tomatoes. Transgenic tomato plants (Lycopersicon esculentum Mill cv. Ailsa Craig) with reduced exo-galactanase/beta-galactosidase mRNA were generated to test this hypothesis and to investigate the role of the enzyme in fruit softening. A previously identified tomato beta-galactosidase cDNA clone, TBG1, was used in the experiments. Heterologous expression of the clone in yeast demonstrated that TBG1 could release galactosyl residues from tomato cell wall galactans. Transgenic plants showed a reduction in TBG1 mRNA to 10% of normal levels in the ripening fruits. However, despite the reduction in message, total beta-galactosidase and exo-galactanase activities were unaffected. Furthermore, there was no apparent effect on levels of cell wall galactosyl residues when compared with the control. It was concluded that during the ripening of tomato fruits a family of beta galactosidases capable of degrading cell wall galactans are active and down regulation of TBG1 message to 10% was insufficient to alter the degree of galactan degradation. PMID- 11413203 TI - Characterization of plasma membrane domains enriched in lipid metabolites. AB - A subpopulation of plasma membrane vesicles enriched in membrane lipid metabolites has been isolated from petals of carnation flowers and leaves of canola seedlings. This was achieved by immunopurification from a microsomal membrane preparation using region-specific antibodies raised against a recombinant polypeptide of the plasma membrane H(+)-ATPase. The properties of this subpopulation of vesicles were compared with those of purified plasma membrane isolated by partitioning in an aqueous dextran-polyethylene glycol two phase system. The lipid composition of the immunopurified vesicles proved to be clearly distinguishable from that of phase-purified plasma membrane, indicating that they represent a unique subpopulation of plasma membrane vesicles. Specifically, the immunopurified vesicles are highly enriched in lipid metabolites, including free fatty acids, diacylglycerol, triacylglycerol and steryl and wax esters, by comparison with the phase-purified plasma membrane. These findings can be interpreted as indicating that lipid metabolites generated within the plasma membrane effectively phase-separate by moving laterally through the plane of the membrane to form discrete domains within the bilayer. It is also apparent that these domains, once formed, are released as vesicles into the cytosol, presumably by microvesiculation from the surface of the plasmalemma. Such removal may be part of normal membrane turnover. PMID- 11413204 TI - Elicitor-induced changes in isoflavonoid metabolism in red clover roots. AB - When roots of 5-d-old red clover (Trifolium pratense L.) seedlings were treated with chitohexaose and CuCl(2), constitutive glucosidic conjugates of formononetin (F) and (-)-maackiain (Ma) promptly disappeared. Free F and Ma, which were not detected in the control tissues, rapidly appeared to reach the maximum levels 24 h after the initiation of treatment and then declined. The pattern of appearance and disappearance was the same between the tissues treated with chitohexaose and CuCl(2). The enzyme activities related to isoflavonoid metabolism were investigated using crude extracts from elicitor-treated roots. The conjugate forming glucosyltransferase and malonyltransferase activities were lost or markedly reduced after elicitor treatment. On the other hand, malonylesterase and glucosidase activities remained unchanged or showed only slight increase. Phenylalanine ammonia-lyase activity disappeared following elicitor treatment. These results indicated that free aglycones were produced from the conjugate pool by hydrolysis under conditions in which the biosynthetic pathway was extinguished. The amount of Ma produced did not explain that of MaGM lost (about 45%). Since Ma, but not its conjugates, served as a substrate for peroxidase from the elicitor-treated roots, Ma was considered to be converted to insoluble materials. PMID- 11413205 TI - Zea mays CCaMK: autophosphorylation-dependent substrate phosphorylation and down regulation by red light. AB - The role of protein kinases has been extensively studied in various signal transduction pathways and they are one of the most important components that link the signal perception to the final response. However, not many studies have been reported, especially from the plant systems, that show the regulation pattern of the kinase itself under different conditions. A calcium/calmodulin-dependent protein kinase has already been purified and characterized from etiolated maize coleoptiles. In this paper a detailed study of how the kinase itself is regulated at the autophosphorylation level is provided. Evidence is also given that the autophosphorylation of kinase effects its activity towards substrate phosphorylation. It is further shown that the kinase is an important component of the light signalling pathway as the level of the kinase itself decreases by red light irradiation. PMID- 11413206 TI - Changes in oligosaccharide content and antioxidant enzyme activities in developing bean seeds as related to acquisition of drying tolerance and seed quality. AB - Seeds of bean (Phaseolus vulgaris cv. Vernel) were collected throughout their development on the plant and dried at 15 degrees C and 75% relative humidity to a final moisture content of about 16% (fresh weight basis) to determine whether the onset of tolerance to this drying condition was related to changes in soluble sugars or the activities of the main antioxidant enzymes, namely superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR). Measurements of soluble sugars and enzyme activities were made after drying the seeds, and drying tolerance was evaluated by the ability of dried seeds to germinate and to produce normal seedlings. Seeds became tolerant to drying at 45 d after anthesis, a time marking physiological maturity. At physiological maturity, the moisture content of seeds was about 50-55% (fresh weight basis) and seed dry matter reached about 190 mg per seed. Seed vigour, evaluated by controlled deterioration and conductivity measurements, continued to increase after seed mass maturity, but decreased when seeds remained thereafter for more than 7 d on the plant. Acquisition of drying tolerance was coincident with an accumulation of raffinose and stachyose. Dried-tolerant seeds were also characterized by a high amount of sucrose, the most abundant sugar, and by a low content of monosaccharides. The (raffinose+stachyose)/sucrose ratio increased during seed filling, reaching a value close to 1 when all the seeds became tolerant to drying, and maintaining this proportion during the final stages of maturation. Acquisition of drying tolerance was also related to a reorientation of the enzymatic antioxidant defence system. Drying-tolerant dried seeds displayed high CAT and GR activities and low SOD and APX activities, while the opposite condition was observed in immature dried seeds. The shift in antioxidant enzymes corresponded to the beginning of the maturation-drying phase. These results suggest that oligosaccharide metabolism and enzymatic antioxidant defences may be involved in acquisition of drying tolerance during bean seed development, but are not related to seed vigour. PMID- 11413207 TI - Bundle sheath cells of small veins in maize leaves are the location of uptake from the xylem. AB - Rb(+) as a tracer for K(+) was used to test the hypothesis that uptake of K(+) from xylem vessels of small veins into the symplast of maize leaves occurs at the xylem/bundle sheath cell interface. 22.5 min after immersing cut leaves into 20 mM RbCl+1 mM KCl, Rb(+) appeared in the cells of the leaves. Sections of these leaves were freeze-dried. In cryo-thin sections (5 microm), (85)Rb(+) and (41)K(+) content was determined by laser microprobe mass analysis with a large resolution of about 1 microm. Determining the ratio of (85)Rb(+) to (41)K(+) in the cell walls and cytosols of bundle sheath cells, mesophyll cells, and in the cells between the xylem elements resulted in the following picture: In small veins, Rb(+) entered the symplast directly at the xylem/bundle sheath cell interface. PMID- 11413208 TI - In vitro control of floral transition in tomato (Lycopersicon esculentum Mill.), the model for autonomously flowering plants, using the late flowering uniflora mutant. AB - In vitro control of floral transition in tomato (Lycopersicon esculentum Mill.), the model plant for autonomously flowering species has been investigated using the late flowering mutant uniflora (uf). Apices collected from truly vegetative plants were cultivated on solid media supplemented with different combinations of growth regulators and chemicals. Several chemical factors implicated in the promotion of floral transition of the uf mutant have been identified: sucrose, cytokinins and nitrogenous nutrients have all to be supplied at optimal concentrations. In contrast, gibberellic acid was found to be inhibitory. These results are discussed in relation to knowledge accumulated on the nature of the flowering signals circulating, at floral transition, in other plants, especially in photoperiodic species. This study suggests that tomato could constitute an adequate model to investigate the genetic and physiological control of floral transition and contribute in unravelling pathways which are constitutively regulating this important step of plant life cycle. PMID- 11413209 TI - Reduced de-etiolation of hypocotyl growth in a tomato mutant is associated with hypersensitivity to, and high endogenous levels of, abscisic acid. AB - A recessive single gene mutant, 7B-1, in tomato was originally selected for its photoperiod-dependent male sterility. The 7B-1 mutant also has some pleiotropic effects including reduced light-induced inhibition, i.e. de-etiolation, of the hypocotyl in long days (LD), increased seed size and weight, and reduced transpiration rate. These traits led us to investigate the sensitivity of 7B-1 to exogenous hormones and the interaction of these responses with daylength. In LD, but not in short days (SD), 7B-1 was more sensitive than wild-type (WT) to exogenous abscisic acid (ABA) for inhibition of seed germination, root elongation and transpiration rate. 7B-1 mutant also exhibited reduced responses to exogenous gibberellin (GA(3)) for hypocotyl elongation, and to inhibitors of GA biosynthesis for seed germination and root and hypocotyl elongation. 7B-1 hypocotyls contained a higher level of endogenous ABA than WT in both photoperiods, although ABA levels were higher in LD than in SD. In contrast, growth-active GAs, i.e. GA(1), GA(3) and GA(4), and IAA were low in the mutant hypocotyls. The 7B-1 mutant appears to be an ABA-overproducer, and the photoperiod-regulated ABA levels may be responsible for the hypersensitivity of the mutant to exogenous ABA. PMID- 11413210 TI - Metabolic inhibition of root water flow in red-osier dogwood (Cornus stolonifera) seedlings. AB - The short-term effects of sodium azide (NaN(3)) on water flow in red-osier dogwood (Cornus stolonifera Michx.) seedlings were examined in excised roots at a constant pressure of 0.3 MPa. NaN(3) significantly decreased root water flow rates (Q(v)). It also induced a significant reduction in root respiration and reduced stomatal conductance to a greater extent in intact seedlings than in excised shoots. Apoplastic flow of water increased with the NaN(3)-induced decreases in Q(v). Mercuric chloride (HgCl(2)) was also used to characterize the water flow responses and respiration of dogwood roots. Similarly to NaN(3), 0.1 and 0.3 mM HgCl(2) decreased root respiration rates and Q(v). The lower, 0.05 mM HgCl(2) treatment, reduced Q(v), but had no significant effect on root oxygen uptake. The reduction of Q(v) in HgCl(2)-treated plants was only partly reversed by 50 mM mercaptoethanol. The mercurial inhibition of Q(v) suggested the presence of Hg-sensitive water channels in dogwood roots. The results indicate that root absorbed NaN(3) metabolically inhibited water channel activities in roots and in shoots and resulted in stomatal closure. It is suggested that the inhibition of respiration that occurs in plants stressed with environmental factors such as flooding, cold soils, and drought may be responsible for the closure of water channels in root cells and inhibition of root water flow. PMID- 11413211 TI - Further observations on the interaction between sugar cane and Gluconacetobacter diazotrophicus under laboratory and greenhouse conditions. AB - Sugar cane (Saccharum spp.) variety SP 70-1143 was inoculated with Gluconacetobacter diazotrophicus strain PAL5 (ATCC 49037) in two experiments. In experiment 1 the bacteria were inoculated into a modified, low sucrose MS medium within which micropropagated plantlets were rooted. After 10 d there was extensive anatomical evidence of endophytic colonization by G. diazotrophicus, particularly in lower stems, where high numbers of bacteria were visible within some of the xylem vessels. The identity of the bacteria was confirmed by immunogold labelling with an antibody raised against G. diazotrophicus. On the lower stems there were breaks caused by the separation of the plantlets into individuals, and at these 'wounds' bacteria were seen colonizing the xylem and intercellular spaces. Bacteria were also occasionally seen entering leaves via damaged stomata, and subsequently colonizing sub-stomatal cavities and intercellular spaces. A localized host defence response in the form of fibrillar material surrounding the bacteria was associated with both the stem and leaf invasion. In experiment 2, stems of 5-week-old greenhouse-grown plants were inoculated by injection with a suspension of G. diazotrophicus containing 10(8) bacteria ml(-1). No hypersensitive response (HR) was observed, and no symptoms were visible on the leaves and stems for the duration of the experiment (7 d). Close to the point of inoculation, G. diazotrophicus cells were observed within the protoxylem and the xylem parenchyma, where they were surrounded by fibrillar material that stained light-green with toluidine blue. In leaf samples taken up to 4 cm from the inoculation points, G. diazotrophicus cells were mainly found within the metaxylem, where they were surrounded by a light green-staining material. The bacteria were growing in relatively low numbers adjacent to the xylem cell walls, and they were separated from the host-derived material by electron-transparent 'haloes' that contained material that reacted with the G. diazotrophicus antibody. PMID- 11413212 TI - Involvement of ethylene in the maturation of black spruce embryogenic cell lines with different maturation capacities. AB - To examine the possible relationship between ethylene and the capacity of embryogenic cell lines to produce mature somatic embryos of black spruce (Picea mariana (Mill.) B.S.P.), two embryogenic cell lines which exhibit different maturation capacities were used to analyse ethylene biosynthesis and that of its immediate precursor, 1-aminocyclopropane-1- carboxylic acid (ACC). Several compounds known to alter ethylene metabolism were also evaluated for their effect on the number of mature somatic embryos produced. The results showed that in the high capacity cell line, ethylene production and endogenous ACC pools were less than in the low capacity cell line. It was also demonstrated that limiting ethylene biosynthesis by adding inhibitors of ethylene biosynthesis or its physiological action to the maturation medium promoted somatic embryo production for the low capacity cell line. Conversely, lowering ethylene biosynthesis reduced the number of somatic embryos in the high capacity cell line. These results were further substantiated by the finding that the effects of amino oxyacetic acid (AOA), an inhibitor of ethylene biosynthesis, were partially reversed by adding ethylene to both embryogenic cell lines. It is concluded that ethylene is implicated in somatic embryogenesis of black spruce and that the low capacity cell line had excess, i.e. supraoptimal, ethylene production, whereas the high capacity cell line had nearly optimal ethylene production. The relationship between ethylene and other phytohormones, and the possible effects of the interaction between ethylene and polyamines on the maturation of the somatic embryos are discussed. PMID- 11413213 TI - Quantification of stomatal uptake of ionic solutes using a new model system. AB - Evidence for stomatal uptake of solutes by leaves without the application of surfactants or pressure has recently been provided (Eichert et al., 1998). In the present study, experimental conditions were refined in that the water potential was held at <0 on the physiologically inner side of the epidermal strips (ES) by use of a ceramic plate or the proper mesophyll of the plant. The penetrated substances were immobilized on the inner side by ion exchange membranes. The influence of humidity, light, stomatal density, and re-wetting on the uptake of anions (fluorescein) and cations (Fe(3+)) was investigated, using leaves of Allium porrum, Commelina communis and Sedum telephium. Uptake increased with humidity, stomatal aperture and stomatal density. It was restricted to stomatal areas, and was especially high below the rims of drying droplets. Again, penetration of stomatal pores was observed. Uptake was strongly correlated with the number of penetrated stomata, although usually less than 10% of the stomata contributed to uptake. The number of stomata that had been penetrated was highly variable, increasing extremely significantly with the number of repeated drying/ wetting cycles. These results indicate that stomatal uptake can be a major pathway for the foliar uptake of ionic solutes. It is a dynamic process, depending on environmental conditions and history of the residues on the leaf, aspects that had been neglected in previous concepts. PMID- 11413214 TI - Intra-specific variation in xylem cavitation in interior live oak (Quercus wislizenii A. DC.). AB - Xylem cavitation induced by water stress reduces plant hydraulic conductance and can indicate the habitat a species evolved in and its phylogenetic background. Species differ widely in cavitation resistance, but less is known about intra specific variation. Cavitation resistance was assessed for field-collected adult and sapling size classes from three populations of interior live oak (Quercus wislizenii A. DC.) in California, USA. Root and stem cavitation resistance of two year old seedlings from a greenhouse experiment was also measured. Cavitation resistance curves were determined by injecting air into the vascular system to induce cavitation and measuring the subsequent decline in hydraulic conductance. Based on the air-seeding hypothesis, the absolute value of the air pressures should be equivalent to the tensions that cause cavitation under dehydrating conditions. Conductance declined exponentially with applied pressure for both roots and stems. Comparisons between populations did not reveal significant differences despite good statistical power. The 50% loss in conductance point occurred between 1.0-1.6 MPa; conductance declined more slowly thereafter. Conductance was 21-30% of maximum at 4.0 MPa and 7-14% at 8.0 MPa. Saplings exhibited a nearly identical pattern compared with adults except at 4.0 MPa, where saplings exhibited slightly less cavitation (7%). Greenhouse seedling stems were more resistant compared with both field-collected adults and with seedling roots. The 50% loss in conductance point occurred at 0.83 and 2.6 MPa for seedling roots and stems, respectively. Seedling stems maintained conductance of 20.9% at 8.0 MPa while most roots were fully cavitated between 5.0-8.0 MPa. PMID- 11413215 TI - Why plants grow poorly on very acid soils: are ecologists missing the obvious? AB - Factors associated with soil acidity are considered to be limiting for plants in many parts of the world. This work was undertaken to investigate the role of the toxicity of hydrogen (H(+)) which seems to have been underconsidered by ecologists as an explanation of the reduced plant growth observed in very acid soils. Racial differences are reported in plant growth response to increasing acidity in the grass Holcus lanatus L. (Yorkshire-fog) and the tree Betula pendula Roth (Silver Birch). Soils and seeds were collected from four Scottish sites which covered a range of soils from acid (organic and mineral) to more base rich. The sites and their pH (1:2.5 fresh soil:0.01 M CaCl(2)) were: Flanders Moss (FM), pH 3.2+/-0.03; Kippenrait Glen (KP), pH 4.8+/- 0.05; Kinloch Rannoch (KR), pH 6.1+/-0.16; and Sheriffmuir (SMM), pH 4.3+/-0.11. The growth rates of two races of H. lanatus, FM and KP, and three races of B. pendula (SMM, KP and KR) were measured in nutrient solution cultures at pH 2.0 (H. lanatus only), 3.0, 4.0, 5.0, and 5.6. Results showed races from acid organic soils (FM) were H(+) tolerant while those from acid mineral soils (SMM) were Al(3+)-tolerant but not necessarily H(+)-tolerant. These results confirmed that populations were separately adapted to H(+) or Al(3+) toxicity and this was dependent upon the soil characteristics at their site of collection. The fact of plant adaptation to H(+) toxicity supports the view that this is an important factor in very acid soils. PMID- 11413216 TI - The high oxygen atmosphere toward the end-Cretaceous; a possible contributing factor to the K/T boundary extinctions and to the emergence of C(4) species. AB - Angiosperm plants were grown under either the present day 21 kPa O(2) atmosphere or 28 kPa, as estimated for the end-Cretaceous (100-65 MyBP). CO(2) was held at different levels, within the 24-60 Pa range, as also estimated for the same period. In C(3) Xanthium strumarium and Atriplex prostrata, leaf area and net photosynthesis per unit leaf area, were reduced by the high O(2), while the whole plant respiration/photosynthesis ratio increased. The high O(2) effects were strongest under 24 Pa, but still significant under 60 Pa CO(2). Growth was reduced by high O(2) in these C(3) species, but not in Flaveria sp., whether C(3), C(4), or intermediary grown under light intensities <350 micromol m(-2) s( 1) PPF. Photosynthesis of C(3) Flaveria sp. was reduced by high O(2), but only at light intensities >350 micromol m(-2) s(-1) PPF. It is concluded that the high O(2) atmosphere at the end-Cretaceous would have reduced growth of at least some of the vegetation, thus adversely affecting dependent fauna. The weakened biota would have been predisposed to the consequences of volcanism and the K/T boundary bolide impact. Conversely, photosynthesis and growth of C(4) Zea mays and Atriplex halimus were little affected by high, 28 kPa, O(2). This suggests an environmental driver for the evolution of C(4) physiology. PMID- 11413217 TI - Responses to bleaching herbicides by leaf chloroplasts of maize plants grown at different temperatures. AB - The effects of growth temperature on chloroplast responses to norflurazon and amitrole, two herbicides inhibiting carotenogenesis, at phytoene desaturation and lycopene cyclization, respectively, were studied in leaves of maize plants grown at 20 degrees C and 30 degrees C in light. At the lower temperature both chemicals caused severe photo-oxidative damage to chloroplasts. In organelles of norflurazon-treated leaves neither carotenoids nor chlorophylls were detectable and the thylakoid system was dismantled. In organelles of amitrole-treated leaves lycopene was accumulated, but small quantities of beta-carotene and xanthophylls were also produced. Moreover, some chlorophyll and a few inner membranes still persisted, although these latter were disarranged, lacking essential protein components and devoid of photosynthetic function. The increase in plant growth temperature to 30 degrees C did not change the norflurazon effects on carotenoid synthesis and the photo-oxidative damage suffered by chloroplasts. By contrast, in organelles of amitrole-treated leaves a large increase in photoprotective carotenoid biosynthesis occurred, with a consequent recovery of chlorophyll content, ultrastructural organization and thylakoid composition and functionality. This suggests that thermo-modulated steps could exist in the carotenogenic pathway, between the points inhibited by the two herbicides. Moreover it shows that, unlike C(3) species, C(4) species, such as maize, can express a strong tolerance to herbicides like amitrole, when supplied to plants growing at their optimum temperature conditions. PMID- 11413218 TI - Towards optimization of growth via nutrient supply phasing: nitrogen supply phasing increases broccoli (Brassica oleracea var. italica) growth and yield. AB - A greenhouse experiment on broccoli (Brassica oleracea var. italica, cvs Windsor and Arcadia) was carried out in order to demonstrate that supplying nitrogen (N) to meet the nitrogen demands of plant growth stages, through N phasing, improves plant growth and yield, as compared to fertilizing at the conventional, optimal, constant N rate. Two broccoli cultivars and two rates of starter nitrogen fertilizer (optimum, 250 mg l(-1) and sub-optimum, 150 mg l(-1)), were combined with three timings of fertigation change. Shifting N rate, at 60% and 75% of the market plant growth cycle significantly increased shoot dry weight and head fresh weight, compared to the constant-N rates treatments (controls). The highest yield and shoot dry weight were obtained when the N-rate was switched from the optimum level (250 mg l(-1)) to the sub-optimum level (150 mg l(-1)) at inflorescence initiation. The nitrogen-to-growth-stage-fitness effect was determined and partitioned into rate effect and phasing effect. The phasing effect was greatest, on both shoot dry weight and head fresh weight, at inflorescence initiation, and subsequently decreased until harvest time. None of the interactions was significant. The results demonstrated the superiority of nitrogen supply phasing over the conventional fixed-rate-supply method. PMID- 11413219 TI - The response of the high altitude C(4) grass Muhlenbergia montana (Nutt.) A.S. Hitchc. to long- and short-term chilling. AB - The acclimation of C(4) photosynthesis to low temperature was studied in the montane grass Muhlenbergia montana in order to evaluate inherent limitations in the C(4) photosynthetic pathway following chilling. Plants were grown in growth cabinets at 26 degrees C days, but at night temperatures of either 16 degrees C (the control treatment), 4 degrees C for at least 28 nights (the cold-acclimated treatment), or 1 night (the cold-stress treatment). Below a measurement temperature of 25 degrees C, little difference in the thermal response of the net CO(2) assimilation rate (A) was observed between the control and cold-acclimated treatment. By contrast, above 30 degrees C, A in the cold-acclimated treatment was 10% greater than in the control treatment. The temperature responses of Rubisco activity and net CO(2) assimilation rate were similar below 22 degrees C, indicating high metabolic control of Rubisco over the rate of photosynthesis at cool temperatures. Analysis of the response of A to intercellular CO(2) level further supported a major limiting role for Rubisco below 20 degrees C. As temperature declined, the CO(2) saturated plateau of A exhibited large reductions, while the initial slope of the CO(2) response was little affected. This type of response is consistent with a Rubisco limitation, rather than limitations in PEP carboxylase capacity. Stomatal limitations at low temperature were not apparent because photosynthesis was CO(2) saturated below 23 degrees C at air levels of CO(2). In contrast to the response of photosynthesis to temperature and CO(2) in plants acclimated for 4 weeks to low night temperature, plants exposed to 4 degrees C for one night showed substantial reduction in photosynthetic capacity at temperatures above 20 degrees C. Because these reductions were at both high and low CO(2), enzymes associated with the C(4) carbon cycle were implicated as the major mechanisms for the chilling inhibition. These results demonstrate that C(4) plants from climates with low temperature during the growing season can fully acclimate to cold stress given sufficient time. This acclimation appears to involve reversal of injury to the C(4) cycle following initial exposure to low temperature. By contrast, carbon gain at low temperatures generally appears to be constrained by the carboxylation capacity of Rubisco, regardless of acclimation time. The inability to overcome the Rubisco limitation at low temperature may be an inherent limitation restricting C(4) photosynthetic performance in cooler climates. PMID- 11413220 TI - Influx and accumulation of Cs(+) by the akt1 mutant of Arabidopsis thaliana (L.) Heynh. lacking a dominant K(+) transport system. AB - An extensive literature reports that Cs(+), an environmental contaminant, enters plant cells through K(+) transport systems. Several recently identified plant K(+) transport systems are permeable to Cs(+). Permeation models indicate that most Cs(+) uptake into plant roots under typical soil ionic conditions will be mediated by voltage-insensitive cation (VIC) channels in the plasma membrane and not by the inward rectifying K(+) (KIR) channels implicated in plant K nutrition. Cation fluxes through KIR channels are blocked by Cs(+). This paper tests directly the hypothesis that the dominant KIR channel in plant roots (AKT1) does not contribute significantly to Cs(+) uptake by comparing Cs(+) uptake into wild type and the akt1 knockout mutant of Arabidopsis thaliana (L.) Heynh. Wild-type and akt1 plants were grown to comparable size and K(+) content on agar containing 10 mM K(+). Both Cs(+) influx to roots of intact plants and Cs(+) accumulation in roots and shoots were identical in wild-type and akt1 plants. These data indicate that AKT1 is unlikely to contribute significantly to Cs(+) uptake by wild-type Arabidopsis from 'single-salt' solutions. The influx of Cs(+) to roots of intact wild-type and akt1 plants was inhibited by 1 mM Ba(2+), Ca(2+) and La(3+), but not by 10 microM Br-cAMP. This pharmacology resembles that of VIC channels and is consistent with the hypothesis that VIC channels mediate most Cs(+) influx under 'single-salt' conditions. PMID- 11413221 TI - A simple protocol for transient gene expression in ripe fleshy fruit mediated by Agrobacterium. AB - Fleshy fruits represent a very important economic resource and, therefore, they are an ideal target for biotechnological ameliorations. However, because of their physiological and anatomical characteristics, ripe fleshy fruits represent an extremely difficult material for transient gene expression assays aimed at the study of gene promoters in a short time. To this purpose, a fast and efficient Agrobacterium-mediated transient gene expression system was developed for ripe fleshy fruits. A beta-glucuronidase reporter gene interrupted by an intron was used in order to prevent the possible expression of GUS activity by the Agrobacterium cells. The contemporary use of another reporter gene was used to check the transformation efficiency. This method is based on the injection of an Agrobacterium suspension into the fruits, and allows both qualitative and quantitative assays in a wide range of fruits to be carried out. PMID- 11413222 TI - Development of an improved medium for germination of Cajanus cajan (L.) Millsp. pollen in vitro. AB - A simple, reliable medium for pollen germination of Cajanus cajan was developed by modifying Brewbaker and Kwack (BK) medium. Past attempts of C. cajan pollen germination in artificial media were not successful. A medium containing polyethylene glycol 4000 (PEG) showed more than 90% germination for C. cajan var. Pusa 33 only when the young buds (36 h before anthesis) were kept in pollen germination medium (PGM) for 36 h before pollen extraction. Supplementation of PGM with epsilon-amino caproic acid (EACA), an amino acid, showed improved pollen germination in Pusa 33 and also helped to avoid preconditioning of young buds before pollen extraction. It was also observed that there is a genotypic difference in the level of EACA required for in vitro pollen germination. Thus a complete medium for C. cajan genotypes consists of 37.5% sucrose+ 15% PEG 4000+250 mg l(-1) boric acid+300 mg l(-1) calcium nitrate+100 mg l(-1) potassium nitrate+ 200 mg l(-1) magnesium sulphate+1% agar+EACA (0, 100, 250, 500, 750 or 1000 mg l(-1)). PMID- 11413223 TI - Age-dependent transformation frequency in elite wheat varieties. AB - Wheat is a major world crop and as such is a primary target for improvement of agronomic characteristics via genetic engineering. Optimization of transformation is essential in order to overcome the relatively low transformation frequencies encountered with wheat. Transformation of elite wheat varieties is not always successful due to variability in regeneration and transformation frequencies between varieties. In this work, two elite wheat varieties with a relatively high embryogenic capacity were transformed by particle bombardment. A strong correlation between transformation frequency and the age of wheat donor plants was observed in both varieties. The mean transformation frequency rose from 0.7% to 5% when using immature embryos from old and young donor plants, respectively. This was observed in both varieties, the best bombardments achieving up to 7.3% frequency. Using explants at an optimal developmental stage from donor plants grown under environmentally-controlled conditions has improved the reproducibility of transformation efficiency of elite wheat varieties and leads to the production of apparently phenotypically normal, fertile, transgenic plants. PMID- 11413224 TI - Procedures allowing the transformation of a range of European elite wheat (Triticum aestivum L.) varieties via particle bombardment. AB - Ten current European wheat varieties were transformed at efficiencies ranging from 1-17% (mean 4% across varieties) following modifications in particle bombardment and tissue culture procedures. All plants surviving phosphinothricin selection were screened for uidA and bar gene activity, and for the presence of marker gene sequences by PCR analysis. A minimum of 35% plant 'escape' frequency was achieved with selection on 4 mg l(-1) gluphosinate ammonium after shoot initiation. Mean co-transformation frequency with various genes-of-interest was 66%. The estimated number of insertions of the uidA gene in 25 lines were; 1-2 in 32%, 3-5 in 52%, and 6-8 in 16% of lines. In T(1) progenies, marker genes segregated in a Mendelian fashion in 50% of 39 lines analysed, as determined by transgene activity assays. Based on PCR analysis, it appeared that in some lines the occurrence of distorted segregation was due to poor transmission of the transgenes. PMID- 11413225 TI - Isolation of a VP1 homologue from wheat and analysis of its expression in embryos of dormant and non-dormant cultivars. AB - A VP (Viviparous) 1 homologous gene has been cloned from wheat (Triticum aestivumL.). Its expression level was examined in the mature embryos of dormant and non-dormant cultivars. The level of expression was positively correlated with the level of seed dormancy and embryo sensitivity to abscisic acid (ABA). PMID- 11413226 TI - A novel Coptis japonica multidrug-resistant protein preferentially expressed in the alkaloid-accumulating rhizome. AB - A full-length cDNA, Cjmdr1, which belongs to the multidrug-resistant (mdr) gene family, was isolated by nested RT-PCR from alkaloid-producing cultured cells of Coptis japonica. The cDNA is 4192 nucleotides long and has an ORF of 1289 amino acids. Northern analysis of the intact plant showed a clear preference in its expression in the rhizome, where alkaloids are highly accumulated compared to other organs. PMID- 11413227 TI - The relationship between the aggregational state of the amyloid-beta peptides and free radical generation by the peptides. AB - In the present study, we investigated whether or not the amyloid-beta protein (Abeta) peptide itself spontaneously generates free radicals using electron spin resonance (ESR) spectroscopy while also monitoring the aggregational state of Abeta and Abeta-induced cytotoxicity. The present results demonstrated a four line spectrum in the presence of both Abeta40 and Abeta42 with Ntert-butyl-alpha phenylnitrone (PBN), but not in the presence of PBN alone in phosphate-buffered saline (PBS). The fact that the four-line spectrum obtained for the Abeta/PBN in PBS was completely abolished in the presence of the iron-chelating agent Desferal demonstrated the observed four-line spectrum to be iron-dependent. The present study also revealed that either Abeta40 or Abeta42 with PBN in phosphate buffer (PB) did not produce any definite four-line spectrum. Both a thioflavine-T (Th-T) fluorometric assay and circular dichroism (CD) spectroscopy showed the amyloid fibril formation of Abeta in PBS to be much higher than that of Abeta in PB. Moreover, Abeta-induced cytotoxicity assays showed Abeta incubated in PBS to be more cytotoxic than that incubated in PB. These results thus suggest that Abeta associated free radical generation is strongly influenced by the aggregational state of the peptides. PMID- 11413228 TI - Differential role of superoxide and glutathione in S-nitrosoglutathione-mediated apoptosis: a rationale for mild forms of familial amyotrophic lateral sclerosis associated with less active Cu,Zn superoxide dismutase mutants. AB - SH-SY5Y cells transfected with the enzymatically inactive Cu,Zn superoxide dismutase mutant H46R were more resistant to S-nitrosoglutathione (GSNO)-induced apoptosis. Cytochrome c release from mitochondria, caspase 3 activation, p53 up regulation, p21 cleavage and Bcl-2 modulation, all involved in the apoptotic process, were significantly less altered with respect to untransfected cells. The H46R resistance to NO was associated with a higher content of reduced glutathione (GSH) and was abolished by blockage of glutathione synthesis. On the other hand, H46R cells were as sensitive as SH-SY5Y cells to puromycin-induced apoptosis; furthermore, they were more susceptible to apoptosis elicited by the superoxide generating drug paraquat and to cell necrosis provoked by t-butyl hydroperoxide. These results confirm that the level of superoxide dismutase activity is fundamental for protecting cells against oxygen free radical challenge. Its impairment is not detrimental to cells exposed to NO, as long as the overall reducing power represented by GSH is assured. These results are relevant to explain a milder progression of the familial amyotrophic lateral sclerosis disease when associated with the H46R mutation. PMID- 11413229 TI - Schedule of NMDA receptor subunit expression and functional channel formation in the course of in vitro-induced neurogenesis. AB - NE-7C2 neuroectodermal cells derived from forebrain vesicles of p53-deficient mouse embryos (E9) produce neurons and astrocytes in vitro if induced by all trans retinoic acid. The reproducible morphological stages of neurogenesis were correlated with the expression of various NMDA receptor subunits. RT-PCR studies revealed that GluRepsilon1 and GluRepsilon4 subunit mRNAs were transcribed by both non-induced and neuronally differentiated cells. GluRepsilon3 subunit mRNAs were not synthesized by NE-7C2 cells and increased numbers of messages from the GluRepsilon2 gene were detected only after neural network formation. The presence of the GluRzeta1 protein was detected throughout neural induction, whereas retinoic acid-induced neuron formation elevated the amount of exon 21 (C1)- and exon 22 (C2)-containing GluRzeta1 mRNAs and resulted in the appearance of exon 5 (N1)-containing transcripts. NMDA-elicited Ca(2+)-signals were detected only in cells displaying neuronal morphology, but preceding the appearance of synapsin-I immunoreactivity. Our findings demonstrated that, in spite of the presence of subunits necessary for channel formation, functional channels were formed by NE 7C2 cells no sooner than the time of neurite maturation. The data show that the cell line provides a suitable model to analyse the mechanisms involved in NMDA receptor gene expression before the appearance of synaptic communication. PMID- 11413230 TI - Glutamate receptor subunit expression in primary neuronal and secondary glial cultures. AB - We report on the expression of ionotropic glutamate receptor subunits in primary neuronal cultures from rat cortex, hippocampus and cerebellum and of metabotropic glutamate (mGlu) receptor subtypes in these neuronal cultures as well as in cortical astroglial cultures. We found that the NMDA receptor (NR) subunits NR1, NR2A and NR2B were expressed in all three cultures. Each of the three cultures showed also expression of the four AMPA receptor subunits. Although RT-PCR detected mRNA of all kainate (KA) subunits in the three cultures, western blot showed only expression of Glu6 and KA2 receptor subunits. The expression analysis of mGlu receptors indicated the presence of all mGlu receptor subtype mRNAs in the three neuronal cultures, except for mGlu2 receptor mRNA, which was not detected in the cortical and cerebellar culture. mGlu1a/alpha, -2/3 and -5 receptor proteins were present in all three cultures, whereas mGlu4a and mGlu8a receptor proteins were not detected. Astroglial cultures were grown in either serum-containing or chemically defined medium. Only mGlu5 receptor protein was found in astroglial cultures grown in serum-containing medium. When astrocytes were cultured in chemically defined medium, mGlu3, -5 and -8 receptor mRNAs were detected, but at the protein level, still only mGlu5 receptor was found. PMID- 11413231 TI - Selective inhibition of beta(2)-adrenergic receptor-mediated cAMP generation by activation of the P2Y(2) receptor in mouse pineal gland tumor cells. AB - Rhythmic noradrenergic signaling from the hypothalamic clock in the suprachiasmatic nucleus to the pineal gland causes an increase in intracellular cAMP which regulates the circadian fluctuation of melatonin synthesis. The activation of phospholipase C (PLC)-coupled P2Y(2) receptors upon treatment with ATP and UTP exclusively inhibited the isoproterenol-stimulated cAMP production in mouse pineal gland tumor cells. However, the activation of other PLC-coupled receptors including P2Y(1) and bombesin receptors had little or no effect on the isoproterenol-stimulated cAMP production. Also, ATP did not inhibit cAMP production caused by forskolin, prostaglandin E(2), or the adenosine analog NECA. These results suggest a selective coupling between signalings of P2Y(2) and beta(2)-adrenergic receptors. The binding of [(3)H]CGP12177 to beta(2)-adrenergic receptors was not effected by the presence of ATP or UTP. Ionomycin decreased the isoproterenol-stimulated cAMP production, whereas phorbol 12-myristate 13-acetate slightly potentiated the isoproterenol response. Chelation of intracellular Ca(2+), however, had little effect on the ATP-induced inhibition of cAMP production, while it completely reversed the ionomycin-induced inhibition. Treatment of cells with pertussis toxin almost completely blocked the inhibitory effect of nucleotides. Pertussis toxin also inhibited the nucleotide-induced increase in intracellular Ca(2+) and inositol 1,4,5-trisphosphate production by 30-40%, suggesting that the ATP-mediated inhibition of the cAMP generation and the partial activation of PLC are mediated by pertussis toxin-sensitive G(i) protein. We conclude that one of the functions of P2Y(2) receptors on the pineal gland is the selective inhibition of beta-adrenergic receptor-mediated signaling pathways via the inhibitory G-proteins. PMID- 11413232 TI - Cytokine-induced cell death in immortalized Schwann cells: roles of nitric oxide and cyclic AMP. AB - Tumor necrosis factor-alpha and interferon-gamma are pleiotropic cytokines that regulate Schwann cell responses during injury and inflammatory demyelination. We have previously shown that cyclic AMP (cAMP)-elevating agents decrease the demyelination and Wallerian degeneration in experimental allergic neuritis. In this study, we examined the role of cAMP in cytokine-mediated signaling in a spontaneously immortal Schwann cell clone (iSC). We found that tumor necrosis factor-alpha and interferon-gamma exert synergistic inhibitory action on Schwann cell viability via the production of nitric oxide (NO) and ceramide (cer). Furthermore, we found that: (i) NO synthase inhibitors attenuate the cytokine induced cer accumulation and cell death indicating that NO acts upstream of cer; and (ii) cytokine-induced cell death is decreased in iSCs pretreated continuously for 48-72 h with forskolin, an activator of adenylate cyclase. Although forskolin modulates the phosphorylation of ERKs and Akt, it decreases the susceptibility of iSC to cytokines via a separate mechanism operating after NO induction and before cer accumulation. We propose that the protective effect of cAMP-elevating agents in experimental allergic neuritis may be mediated in part via modulation of Schwann cell responses to cytokines. PMID- 11413233 TI - Hydrogen peroxide removal and glutathione mixed disulfide formation during metabolic inhibition in mesencephalic cultures. AB - Compromised mitochondrial energy metabolism and oxidative stress have been associated with the pathophysiology of Parkinson's disease. Our previous experiments exemplified the importance of GSH in the protection of neurons exposed to malonate, a reversible inhibitor of mitochondrial succinate dehydrogenase/complex II. This study further defines the role of oxidative stress during energy inhibition and begins to unravel the mechanisms by which GSH and other antioxidants may contribute to cell survival. Treatment of mesencephalic cultures with 10 microM buthionine sulfoximine for 24 h depleted total GSH by 60%, whereas 3 h exposure to 5 mM 3-amino-1,2,4-triazole irreversibly inactivated catalase activity by 90%. Treatment of GSH-depleted cells with malonate (40 mM) for 6, 12 or 24 h both potentiated and accelerated the time course of malonate toxicity, however, inhibition of catalase had no effect. In contrast, concomitant treatment with buthionine sulfoximine plus 3-amino-1,2,4-triazole in the presence of malonate significantly potentiated toxicity over that observed with malonate plus either inhibitor alone. Consistent with these findings, GSH depletion enhanced malonate-induced reactive oxygen species generation prior to the onset of toxicity. These findings demonstrate that early generation of reactive oxygen species during mitochondrial inhibition contributes to cell damage and that GSH serves as a first line of defense in its removal. Pre-treatment of cultures with 400 microM ascorbate protected completely against malonate toxicity (50 mM, 12 h), whereas treatment with 1 mM Trolox provided partial protection. Protein-GSH mixed disulfide formation during oxidative stress has been suggested to either protect vulnerable protein thiols or conversely to contribute to toxicity. Malonate exposure (50 mM) for 12 h resulted in a modest increase in mixed disulfide formation. However, exposure to the protective combination of ascorbate plus malonate increased membrane bound protein-GSH mixed disulfides three-fold. Mixed disulfide levels returned to baseline by 72 h of recovery indicating the reversible nature of this formation. These results demonstrate an early role for oxidative events during mitochondrial impairment and stress the importance of the glutathione system for removal of reactive oxygen species. Catalase may serve as a secondary defense as the glutathione system becomes limiting. These findings also suggest that protein-GSH mixed disulfide formation under these circumstances may play a protective role. PMID- 11413234 TI - Bax kappa, a novel Bax splice variant from ischemic rat brain lacking an ART domain, promotes neuronal cell death. AB - Bax is a pro-apoptotic Bcl-2 family protein that regulates programmed cell death through homodimerization and through heterodimerization with Bcl-2. Bax alpha is encoded by six exons and undergoes alternative splicing. Bax kappa, a splice variant of Bax with conserved BH1, BH2 and BH3 binding domains and a C-terminal transmembrane domain (TM), but with an extra 446-bp insert between exons 1 and 2 leading to loss of an N-terminal ART domain, was identified from an ischemic rat brain cDNA library. Expression of Bax kappa mRNA and protein was up-regulated in hippocampus after cerebral ischemic injury. The increased Bax kappa mRNA was distributed mainly in selectively vulnerable hippocampal CA1 neurons that are destined to die after global ischemia. Overexpression of Bax kappa protein in HN33 mouse hippocampal neuronal cells induced cell death, which was partially abrogated by co-overexpression of Bcl-2. Moreover, co-overexpression of Bax kappa and Bax alpha increased HN33 cell death. The results suggest that the Bax kappa may have a role in ischemic neuronal death. PMID- 11413235 TI - Dual coupling of opioid receptor-like (ORL1) receptors to adenylyl cyclase in the different layers of the rat main olfactory bulb. AB - The coupling of opioid receptor-like (ORL1) receptors to adenylyl cyclase has been investigated in specific layers of the rat main olfactory bulb. Membranes prepared from the olfactory nerve-glomerular layer (ON-G layer), external plexiform layer (EP layer) and granule cell layer (GR layer) displayed specific binding sites for [(3)H]-nociceptin/orphanin FQ ([(3)H]Noc/OFQ). In each layer, the presence of high-and low-affinity binding sites, with K(D) values in the picomolar and nanomolar range, respectively, was detected. The binding of [(3)H]Noc/OFQ was displaced by unlabelled Noc/OFQ, but not by opioid antagonists. In each layer, Noc/OFQ significantly stimulated [(35)S]GTPgammaS binding with nanomolar potencies. In ON-G layer, Noc/OFQ inhibited basal adenylyl cyclase activity and the enzyme stimulations by corticotropin releasing hormone (CRH), Ca(2+)/calmodulin (Ca(2+)/CaM) and forskolin (FSK). In EP layer, Noc/OFQ inhibited Ca(2+)/CaM-and FSK-stimulated enzyme activities. Conversely, in GR layer the peptide stimulated basal cyclase activity and potentiated the enzyme activation by CRH. The Noc/OFQ stimulation was counteracted by the GDP-bound form of the alpha subunit of transducin and was mimicked by transducin betagamma subunits. In the same tissue layer, Ca(2+)/CaM-and FSK-stimulated enzyme activities were inhibited. Naloxone failed to antagonize all the actions of Noc/OFQ. Western blot and RT-PCR analysis revealed the expression of Ca(2+) insensitive and -sensitive adenylyl cyclases in the three layers. These results demonstrate that in rat main olfactory bulb ORL1 receptors can differentially affect distinct forms of adenylyl cyclase in a layer specific manner. PMID- 11413236 TI - Cleavage of Bax is mediated by caspase-dependent or -independent calpain activation in dopaminergic neuronal cells: protective role of Bcl-2. AB - Two cysteine protease families, caspase and calpain, are known to participate in cell death. We investigated whether a stress-specific protease activation pathway exists, and to what extent Bcl-2 plays a role in preventing drug-induced protease activity and cell death in a dopaminergic neuronal cell line, MN9D. Staurosporine (STS) induced caspase-dependent apoptosis while a dopaminergic neurotoxin, MPP(+) largely induced caspase-independent necrotic cell death as determined by morphological and biochemical criteria including cytochrome c release and fluorogenic caspase cleavage assay. At the late stage of both STS- and MPP(+) induced cell death, Bax was cleaved into an 18-kDa fragment. This 18-kDa fragment appeared only in the mitochondria-enriched heavy membrane fraction of STS-treated cells, whereas it was detected exclusively in the cytosolic fraction of MPP(+) treated cells. This proteolytic cleavage of Bax appeared to be mediated by calpain as determined by incubation with [(35)S]methionine-labelled Bax. Thus, cotreatment of cells with calpain inhibitor blocked both MPP(+)- and STS-induced Bax cleavage. Intriguingly, overexpression of baculovirus-derived inhibiting protein of caspase, p35 or cotreatment of cells with caspase inhibitor blocked STS- but not MPP(+)-induced Bax cleavage. This appears to indicate that calpain activation may be either dependent or independent of caspase activation within the same cells. However, cotreatment with calpain inhibitor rescued cells from MPP(+)-induced but not from STS-induced neuronal cell death. In these paradigms of dopaminergic cell death, overexpression of Bcl-2 prevented both STS- and MPP(+)-induced cell death and its associated cleavage of Bax. Thus, our results suggest that Bcl-2 may play a protective role by primarily blocking drug-induced caspase or calpain activity in dopaminergic neuronal cells. PMID- 11413237 TI - Endogenous neurotensin down-regulates dopamine efflux in the nucleus accumbens as revealed by SR-142948A, a selective neurotensin receptor antagonist. AB - SR-142948A belongs to the second generation of potent, selective, non-peptide antagonists of neurotensin receptors. It was used to investigate the role of endogenous neurotensin in the regulation of dopamine efflux in the nucleus accumbens and striatum of anaesthetized and pargyline-treated rats. All the data were obtained using in vivo electrochemistry. Electrically evoked (20 Hz, 10 s) dopamine efflux was monitored by differential pulse amperometry, whereas variations in basal (tonic) dopamine efflux were monitored by differential normal pulse voltammetry. Like the first-generation compound SR-48692, SR-142948A did not affect the tonic and evoked dopamine efflux, but dose-dependently enhanced haloperidol (50 microg/kg, i.p.) induced facilitation of the electrically evoked dopamine release in the nucleus accumbens. In contrast to SR-48692, SR-142948A dose-dependently potentiated haloperidol (50 microg/kg, i.p.) induced increase in the basal dopamine level in the nucleus accumbens. This potentiating effect did not appear in the striatum. When dopaminergic and/or neurotensinergic transmissions were modified by a higher dose of haloperidol (0.5 mg/kg, i.p.), apomorphine, amphetamine or nomifensine, SR-142948A pre-treatment affected only the effect of apomorphine on the basal dopamine level in the nucleus accumbens. These results strengthen the hypothesis that endogenous neurotensin could exert a negative control on mesolimbic dopamine efflux. PMID- 11413238 TI - Proteolysis of glutamate receptor-interacting protein by calpain in rat brain: implications for synaptic plasticity. AB - Activation of the calcium-dependent protease calpain has been proposed to be a key step in synaptic plasticity in the hippocampus. However, the exact pathway through which calpain mediates or modulates changes in synaptic function remains to be clarified. Here we report that glutamate receptor-interacting protein (GRIP) is a substrate of calpain, as calpain-mediated GRIP degradation was demonstrated using three different approaches: (i) purified calpain I digestion of synaptic membranes, (ii) calcium treatment of frozen-thawed brain sections, and (iii) NMDA-stimulated organotypic hippocampal slice cultures. More importantly, calpain activation resulted in the disruption of GRIP binding to the GluR2 subunit of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors. Because GRIP has been proposed to function as an AMPA receptor targeting and synaptic-stabilizing protein, as well as a synaptic-organizing molecule, calpain-mediated degradation of GRIP and disruption of AMPA receptor anchoring are likely to play important roles in the structural and functional reorganization accompanying synaptic modifications in long-term potentiation and long-term depression. PMID- 11413239 TI - Developmental changes in the expression of parkin and UbcR7, a parkin-interacting and ubiquitin-conjugating enzyme, in rat brain. AB - Parkin is a product of the Park2 gene the mutation of which causes autosomal recessive juvenile parkinsonism (AR-JP) characterized by selective dopaminergic neuronal death and absence of Lewy bodies. Recently we found that parkin is directly linked to the ubiquitin (Ub)-proteasome pathway as a Ub-protein ligase (E3) collaborating with a Ub-conjugating enzyme (E2) UbcH7. Here we analysed by in situ hybridization the expression of mRNAs for parkin and UbcR7 (rat orthologue of human UbcH7) in the developing rat brain. Parkin mRNA increased in parallel with neuronal maturation, but was unevenly distributed in various brain regions after four postnatal days. The expression pattern of the UbcR7 mRNA was almost identical to that of the parkin mRNA in all cases examined. Both parkin and UbcR7 mRNAs were distributed in neurones but not glial cells. Our findings indicate that parkin is expressed not only in the substantia nigra, but also uniformly in various brain regions in a development-dependent manner. Co expression of UbcR7 with parkin suggests that UbcR7 may interact with parkin in vivo for ubiquitination of yet unidentified target protein(s). PMID- 11413240 TI - Cerebrin prohormone processing, distribution and action in Aplysia californica. AB - The isolation, characterization, and bioactivity in the feeding circuitry of a novel neuropeptide in the Aplysia californica central nervous system are reported. The 17-residue amidated peptide, NGGTADALYNLPDLEKIamide, has been termed cerebrin due to its primary location in the cerebral ganglion. Liquid chromatographic purification guided by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry allowed the isolation of the peptide with purity adequate for Edman sequencing. The cerebrin cDNA has been characterized and encodes an 86 amino acid prohormone that predicts cerebrin and one additional peptide. Mapping using in situ hybridization and immunocytochemistry showed that cerebrin containing neuronal somata are localized almost exclusively in the cerebral ganglion, mostly in the F- and C-clusters. Both immunostaining and mass spectrometry demonstrated the presence of cerebrin in the neurohemal region of the upper labial nerve. In addition, immunoreactive processes were detected in the neuropil of all of the ganglia, including the buccal ganglia, and in some interganglionic connectives, including the cerebral buccal connective. This suggests that cerebrin may also function as a local signaling molecule. Cerebrin has a profound effect on the feeding motor pattern elicited by the command-like neuron CBI-2, dramatically shortening the duration of the radula protraction in a concentration-dependent manner, mimicking the motor-pattern alterations observed in food induced arousal states. These findings suggest that cerebrin may contribute to food-induced arousal in the animal. Cerebrin-like immunoreactivity is also present in Lymnaea stagnalis suggesting that cerebrin-like peptides may be widespread throughout gastropoda. PMID- 11413241 TI - Nicotine-evoked transmitter release from synaptosomes: functional association of specific presynaptic acetylcholine receptors and voltage-gated calcium channels. AB - It has previously been shown that nicotine-evoked dopamine release from rat striatal synaptosomes and nicotine-evoked norepinephrine release from hippocampal synaptosomes are mediated by distinct nicotinic acetylcholine receptor (nAChR) subtypes. In the present study, the functional association of these nicotinic receptors with specific subtypes of voltage-gated calcium channels was examined. Cd(2+) (200 microM), as well as omega-conotoxin MVIIC (5 microM), blocks approximately 85% of nicotine-evoked dopamine release from striatal synaptosomes, indicating a major involvement of calcium channels. Furthermore, the toxin susceptibility suggests that these calcium channels contain alpha(1A) and/or alpha(1B) subunits. Inhibition of nicotine-evoked dopamine release by conotoxins alpha-MII and omega-GVIA is additive and indicates that presynaptic alpha3beta2 nAChRs are functionally coupled to alpha(1A), but not alpha(1B), calcium channel subtypes. Conversely, insensitivity to alpha-AuIB and sensitivity to omega-MVIIC indicate that non-alpha3beta2/alpha3beta4-containing nAChRs are functionally coupled to alpha(1B)-containing calcium channels. In contrast, Cd(2+) blocks only 65% of nicotine-evoked norepinephrine release from hippocampal synaptosomes, indicating that a substantial fraction of this release occurs through mechanisms not involving calcium channels. This Cd(2+)-insensitive component of release is blocked by alpha-AuIB and therefore appears to be triggered by Ca(2+) flowing directly through the channels of presynaptic alpha3beta4 nAChRs. Thus, these data indicate that different presynaptic termini can have distinctive functional associations of specific nAChRs and voltage-gated calcium channels. PMID- 11413242 TI - Inverse agonists and neutral antagonists at mu opioid receptor (MOR): possible role of basal receptor signaling in narcotic dependence. AB - The mu opioid receptor, MOR, displays spontaneous agonist-independent (basal) G protein coupling in vitro. To determine whether basal MOR signaling contributes to narcotic dependence, antagonists were tested for intrinsic effects on basal MOR signaling in vitro and in vivo, before and after morphine pretreatment. Intrinsic effects of MOR ligands were tested by measuring GTPgammaS binding to cell membranes and cAMP levels in intact cells. beta-CNA, C-CAM, BNTX, and nalmefene were identified as inverse agonists (suppressing basal MOR signaling). Naloxone and naltrexone were neutral antagonists (not affecting basal signaling) in untreated cells, whereas inverse agonistic effects became apparent only after morphine pretreatment. In contrast, 6alpha- and 6beta-naltrexol and -naloxol, and 6beta-naltrexamine were neutral antagonists regardless of morphine pretreatment. In an acute and chronic mouse model of morphine-induced dependence, 6beta naltrexol caused significantly reduced withdrawal jumping compared to naloxone and naltrexone, at doses effective in blocking morphine antinociception. This supports the hypothesis that naloxone-induced withdrawal symptoms result at least in part from suppression of basal signaling activity of MOR in morphine-dependent animals. Neutral antagonists have promise in treatment of narcotic addiction. PMID- 11413243 TI - Astrocytes protect neurons from nitric oxide toxicity by a glutathione-dependent mechanism. AB - Nitric oxide (NO) contributes to neuronal death in cerebral ischemia and other conditions. Astrocytes are anatomically well positioned to shield neurons from NO because astrocyte processes surround most neurons. In this study, the capacity of astrocytes to limit NO neurotoxicity was examined using a cortical co-culture system. Astrocyte-coated dialysis membranes were placed directly on top of neuronal cultures to provide a removable astrocyte layer between the neurons and the culture medium. The utility of this system was tested by comparing neuronal death produced by glutamate, which is rapidly cleared by astrocytes, and N-methyl D-aspartate (NMDA), which is not. The presence of an astrocyte layer increased the LD(50) for glutamate by approximately four-fold, but had no effect on NMDA toxicity. Astrocyte effects on neuronal death produced by the NO donors S-nitroso N-acetyl penicillamine and spermine NONOate were examined by placing these compounds into the medium of co-cultures containing either a control astrocyte layer or an astrocyte layer depleted of glutathione by prior exposure to buthionine sulfoximine. Neurons in culture with the glutathione-depleted astrocytes exhibited a two-fold increase in cell death over a range of NO donor concentrations. These findings suggest that astrocytes protect neurons from NO toxicity by a glutathione-dependent mechanism. PMID- 11413244 TI - Expression of cyclin-dependent kinase 5 and its activator p35 in models of induced apoptotic death in neurons of the substantia nigra in vivo. AB - Cyclin-dependent kinase 5 is predominantly expressed in postmitotic neurons and plays a role in neurite elongation during development. It has also been postulated to play a role in apoptosis in a variety of cells, including neurons, but little is known about the generality and functional significance of cdk5 expression in neuronal apoptosis in living brain. We have therefore examined its expression and that of its known activators, p35, p39 and p67, in models of induced apoptosis in neurons of the substantia nigra. We find that cdk5 is expressed in apoptotic profiles following intrastriatal injection of 6 hydroxydopamine and axotomy. It is expressed exclusively in profiles which are in late morphologic stages of apoptosis. In these late stages, derivation of the profiles from neurons, and localization of expression to the nucleus, can be demonstrated by co-labeling with a neuron-specific nuclear marker, NeuN. In another model of induced apoptotic death in nigra, produced by developmental striatal lesion, kinase activity increases in parallel with cell death. While mRNAs for all three cdk5 activators are expressed in nigra during development, only p35 protein is expressed in apoptotic profiles. We conclude that cdk5/p35 expression is a general feature of apoptotic neuron death in substantia nigra neurons in vivo. PMID- 11413245 TI - Coupling of agonist-induced AMPA receptor internalization with receptor recycling. AB - Excitatory post-synaptic currents in the CNS are primarily mediated by alpha amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) receptors in response to glutamate. Internalization of cell-surface receptors has been shown to be one mechanism by which to control receptor function. To test for agonist control of AMPA receptor plasma membrane expression we used biochemical assays to study AMPA receptor internalization and insertion processes. In heterologous cells, we observed a slow constitutive internalization and a rapid agonist-induced internalization of AMPA receptors. To our surprise, however, agonist treatment had no effect on the steady-state levels of AMPA receptors on the cell surface. To examine whether this could be explained by an agonist-induced increase in the insertion rate of AMPA receptors into the plasma membrane we developed an assay to independently measure receptor insertion. Remarkably, agonist treatment of cells also dramatically increased AMPA receptor plasma membrane insertion rates. In addition, using an assay to measure recycling of internalized pools we found that internalized receptors are rapidly recycled to the cell surface. These results suggest that agonist-induced receptor internalization is coupled to increases in receptor recycling. This increase in receptor flux through intracellular pools may allow for rapid changes in receptor surface expression by independent regulatory control of internalization and insertion. PMID- 11413246 TI - Hypo-osmotic swelling-activated release of organic osmolytes in brain slices: implications for brain oedema in vivo. AB - A decrease in the intracellular levels of osmotically active species has invariably been seen after swelling of mammalian brain tissue preparations. The exact identity of the species, and the manner of their decrease, remain to be described. We investigated the swelling-activated decrease of organic osmolytes in rat cortical brain slices using (1)H- and (31)P-magnetic resonance spectroscopy. We found that acute hypo-osmotic shock causes decreases in the levels of a range of intracellular amino acids and amino acid derivatives, N acetyl-aspartate, creatine, GABA, glutamate, hypotaurine, and also in the levels of the methylamines glycerol-phosphorylcholine, phosphorylcholine and choline. Incubation of cortical slices with the anion channel blockers niflumic acid and tamoxifen caused inhibition of organic osmolyte efflux, suggesting that such osmolyte efflux occurs through anion channels. Intracellular phosphocreatine was also seen to decrease during acute hypo-osmotic superfusion, although intracellular ATP remained constant. In addition, the acidification of an intracellular compartment was observed during hypo-osmotic superfusion. Our results suggest a link between brain energy reserve and brain osmoregulation. PMID- 11413247 TI - Basic fibroblast growth factor: a potential inhibitor of glutamine synthetase expression in injured neural tissue. AB - Basic fibroblast growth factor (bFGF) was recently shown to promote the survival of neural cells and tissues, raising hopes for its therapeutic potential in degenerative disorders of the CNS. Here we examine the effect of bFGF on the expression of glutamine synthetase, a key enzyme in the detoxification of the neurotransmitter glutamate. Expression of this enzyme is regulated by systemic glucocorticoids and, in chick neural retina tissue, is restricted to Muller glial cells. We report that exogenous supply of bFGF to retinal explants inhibits hormonal induction of glutamine synthetase expression. This inhibition appears to be mediated by the c-Jun protein which accumulated, in response to bFGF, exclusively in Muller glial cells. Ischemic conditions, which reportedly stimulate the release of endogenous bFGF, also led to an increase in c-Jun protein and a decline in glutamine synthetase expression. This decline could be competitively prevented by a soluble fibroblast growth factor receptor but not by a soluble epidermal growth factor receptor. The finding that endogenous release of bFGF or its exogenous supply down-regulates glutamine synthetase expression suggests that in addition to its reported neuroprotective effect, bFGF may exacerbate glutamate mediated neurotoxicity through direct down-regulation of glutamine synthetase. PMID- 11413248 TI - Absolute quantification of AMPA receptor subunit mRNAs in single hippocampal neurons. AB - alpha-Amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptor subunit (GluR1-4) mRNAs expressed by single neurons in rat hippocampal cultures were quantified by single-cell RT-PCR using an internal standard RNA after whole-cell patch-clamp recording. The internal standard RNA, derived from GluR2 with a single nucleotide substitution, was reverse-transcribed and PCR-amplified with the same efficiency as GluR1-4 mRNAs. The mean mRNA numbers harvested in vitro from pyramidal-like neurons on day 9 were 1150 +/- 324 molecules of GluR1, 1080 +/- 273 molecules of GluR2, 100 +/- 20 molecules of GluR3, and 50 +/- 10 molecules of GluR4 (mean +/- SEM, n = 12). In a non-pyramidal neuronal population that expresses AMPA receptors characterized by high Ca(2+) permeability, the numbers of GluR1, GluR3 and GluR4 mRNA molecules harvested per cell were 354 +/- 64, 25 +/- 17 and 168 +/- 36, respectively (n = 8). The GluR2 mRNA was not detected in this cell type. The calculated ratio of AMPAR mRNA molecules per total mRNA molecules was 1/240 in pyramidal-like neurons (1/500 for GluR2), being in the range obtained with total RNA from rat forebrain and cerebellum (1/170 and 1/380, respectively). Finally, our results indicated that the proportion of GluR1 4 mRNA located in neurites reached approximately 60% in pyramidal-like neurons. However, we found no evidence of preferential subcellular distribution of a given subunit. PMID- 11413249 TI - The vanilloid receptor (VR1)-mediated effects of anandamide are potently enhanced by the cAMP-dependent protein kinase. AB - The endogenous cannabinoid receptor ligand, anandamide (AEA), is a full agonist of the vanilloid receptor type 1 (VR1) for capsaicin. Here, we demonstrate that the potency and efficacy of AEA at VR1 receptors can be significantly increased by the concomitant activation of protein kinase A (PKA). In human embryonic kidney (HEK) cells over-expressing human VR1, AEA induces a rise in cytosolic Ca(2+) concentration that is mediated by this receptor. The EC(50) for this effect was decreased five-fold in the presence of forskolin (FRSK, 1-5 microM) or the cAMP analogue, 8-Br-cAMP (10-100 microM). The effects of 8-Br-cAMP and FRSK were blocked by a selective PKA inhibitor. The FRSK (10 nM) also potently enhanced the sensory neurone- and VR1-mediated constriction by AEA of isolated guinea-pig bronchi, and this effect was abolished by a PKA inhibitor. In rat dorsal root ganglia slices, AEA-induced release of substance P, an effect mediated by VR1 activation, was enhanced three-fold by FRSK (10 nM). Thus, the ability of AEA to stimulate sensory VR1, with subsequent neuropeptide release, appears to be regulated by the state of activation of PKA. This observation supports the hypothesis that endogenous AEA might stimulate VR1 under certain pathophysiological conditions. PMID- 11413250 TI - Enhanced inducible mGlu1alpha receptor expression in Chinese hamster ovary cells. AB - Inducible expression of the group-I metabotropic glutamate receptor (mGlu1alpha) in Chinese hamster ovary cells allows for the study of receptor density dependent effects. However, expression levels attainable with this system are lower than those reported for various brain regions and achieved by conventional (constitutive) transfection. Thus, direct comparison of mGlu1alpha receptor mediated responses in this inducible expression system with those for receptors expressed heterologously or in vivo is compounded. We show here that inducible expression can be selectively augmented by butyrate pretreatment to levels approaching those reported for cerebral tissue. Enhanced mGlu1alpha receptor protein levels, agonist-induced inositol phosphate accumulation, as well as single-cell inositol 1,4,5-trisphosphate production and intracellular Ca(2+) mobilization occurred following co-induction with butyrate. In contrast, endogenous purinoceptor function was unaffected. Importantly, the ability to titrate receptor expression by varying isopropyl beta-thiogalactoside concentration was retained. Sodium butyrate thus offers a simple and convenient method to enhance inducible gene expression to levels found in vivo. PMID- 11413251 TI - Idiopathic intracranial hypertension: any light on the mechanism of the raised pressure? PMID- 11413252 TI - The volumes of memory. PMID- 11413253 TI - Long term effects of locomotor training in spinal humans. PMID- 11413254 TI - Cognitive function in the oldest old: Women perform better than men. PMID- 11413255 TI - Cervical vertigo. PMID- 11413256 TI - Structural MRI volumetric analysis in patients with organic amnesia, 1: methods and comparative findings across diagnostic groups. AB - BACKGROUND: If they are to be replicable, MRI volume measurements require explicit definitions of structures and of criteria for delineating these structures on MRI. Previously published volumes in healthy subjects show considerable differences in measurements across different studies, including a fourfold variation in estimates of hippocampal volume. Previous neuroimaging reports in patients with Korsakoff syndrome have generally found widespread or non-specific change, whereas in patients with herpes encephalitis the extent of pathological involvement reported beyond the temporal lobes has varied. METHOD: In the present study, a clear set of anatomical criteria and detailed MRI segmentation procedures were applied to measure whole brain, frontal and temporal lobe, and anterolateral and medial temporal volumes, as well as thalamic areas in patients with organic amnesia (from Korsakoff's syndrome, herpes encephalitis, and focal frontal lesions) as well as healthy controls. RESULTS: Patients with Korsakoff's syndrome showed decreased thalamic measurements but no significant changes in the medial temporal lobes, whereas patients with herpes encephalitis showed severe medial temporal but not thalamic atrophy. In the patients with known frontal lobe lesions, quantitative analysis on MRI showed reduced frontal lobe volume but no significant temporal lobe or thalamic atrophy. CONCLUSION: Quantified MRI can be a useful technique with which to examine brain-cognitive relations, provided that detailed techniques are explicitly described. In particular, specific patterns of volume change can be found in vivo in patients with Korsakoff's syndrome and those with herpes encephalitis. PMID- 11413257 TI - Structural MRI volumetric analysis in patients with organic amnesia, 2: correlations with anterograde memory and executive tests in 40 patients. AB - BACKGROUND: Cognitive-MRI correlations have often been studied in disorders in which there are multiple cognitive deficits and widespread cortical atrophy, such as Alzheimer's dementia. In such circumstances, the interpretation of any single cognitive-structural correlation is equivocal. Only by measuring differing cognitive functions and a wide range of brain structures in patients with a varying distribution of lesions or atrophy can specific brain-cognitive relations be determined in neurological disorder. METHOD: In the present study, a clear set of anatomical criteria and detailed MRI segmentation procedures were applied to measure whole brain, and left and right frontal, temporal lobe, anterolateral and medial temporal volumes, as well as thalamic cross sectional areas in 40 patients with organic amnesia (from various diseases) and 10 healthy controls. RESULTS: Within the total patient group, anterograde memory measures correlated significantly with medial temporal, hippocampal, and thalamic measurements. A spatial memory measure correlated significantly with hippocampal volume, and temporal context memory with frontal volume. After a factor analysis of the cognitive measures, the association between anterograde memory and hippocampal volume was corroborated. Forgetting rates and subjective memory evaluations did not show any significant MR correlations and, of executive tests employed, only card sorting categories correlated significantly with frontal volume. CONCLUSION: Loss of volume in key brain structures (for example, hippocampus, thalamus) is detectable on quantitative MRI, and this loss of volume correlates significantly with impaired performance on measures of anterograde memory function. Correlations with hippocampal volume did not indicate a specific role in either recall or verbal memory, as opposed to recognition or visual memory. PMID- 11413258 TI - Cognitive function in the oldest old: women perform better than men. AB - OBJECTIVE: Limited formal education is associated with poor cognitive function. This could explain sex differences in cognitive function in the oldest old. Whether limited formal education explains differences in cognitive function between elderly women and men was explored. METHODS: The Leiden 85-plus Study is a population based study investigating all 85 year old inhabitants of Leiden with an overall response rate of 87%. A sample of 599 participants were visited at their place of residence. The mini mental state examination was completed by all participants. Cognitive speed and memory were determined with four neuropsychological tests in participants with a mini mental state examination score higher than 18 points. RESULTS: The proportion of women with limited formal education was significantly higher than that of men (70% v 53%, p=0.001), but women had better scores for cognitive speed and memory than men (p<0.05). After adjustment for differences in limited formal education and the presence of depressive symptoms, the odds ratio for women to have a higher cognitive speed than men was 1.7 (95% CI; 1.0 to 2.6), and for them to have a better memory the odds ratio was 1.8 (95%CI; 1.2 to 2.7). CONCLUSION: Women have a better cognitive function than men, despite their lower level of formal education. Limited formal education alone, therefore, cannot explain the differences in cognitive function in women and men. These findings support the alternative hypothesis that biological differences, such as atherosclerosis, between women and men account for the sex differences in cognitive decline. PMID- 11413259 TI - Patients with Alzheimer's disease and dementia with Lewy bodies mistaken for Creutzfeldt-Jakob disease. AB - OBJECTIVES: To describe the clinical presentation of patients with Alzheimer's disease (AD) or dementia with Lewy bodies (DLB) who were suspected of having Creutzfeldt-Jakob disease (CJD) and to investigate whether current clinical diagnostic criteria cover these atypical forms of AD and DLB. METHODS: Brains from necropsy were examined for the diagnosis of CJD at the German reference centre for spongiform encephalopathies. Symptoms and signs in patients with suspected CJD in whom necropsy showed AD (n=19) or DLB (n=12) were analysed. Their data were compared with a group of patients with CJD (n=25) to determine overlapping and discriminating clinical features. All patients were classified according to clinical diagnostic criteria for CJD, AD, and DLB. RESULTS: Demented patients were suspected of having CJD if disease was rapidly progressing and/or focal neurological signs appeared and/or an EEG showed sharp wave complexes. Myoclonus and limb rigidity were the most common neurological signs in all three dementias. DLB was not suspected in any patient, although patients with DLB showed parkinsonism (58%) and fluctuations (58%). Periodic sharp wave complexes (PSWCs) in EEG typical of CJD were found in five patients with AD and one patient with DLB. 14-3-3 Protein in CSF was detected in 20 patients with CJD, in two patients with AD, but not in any patient with DLB. Although most patients with DLB or AD met the clinical criteria for their respective diagnosis (74% and 90%), they also fulfilled criteria for CJD (42% and 58%). CONCLUSIONS: In patients with rapidly progressive dementia and focal neurological signs, CJD should be the first line diagnosis. Facing the triad dementia, myoclonus, and rigidity, AD should be considered if the disease course is longer and DLB is the differential diagnosis if parkinsonism or fluctuations are present. Findings on EEG or CSF typical of CJD do not exclude AD or DLB. PMID- 11413260 TI - Memory lost, memory regained: neuropsychological findings and neuroimaging in two cases of paraneoplastic limbic encephalitis with radically different outcomes. AB - OBJECTIVE: To report two cases of paraneoplastic limbic encephalitis (PNLE) with similar clinical presentation, but dramatically different outcome and to highlight the role of neuropsychological and radiological evaluation in PNLE. METHODS: Both patients underwent an extensive battery of neuropsychological tests designed to document general intellectual function, anterograde verbal and visual memory, naming, knowledge and executive ability. In addition, structural (CT and MRI) and functional (HMPAO-SPECT) brain scans were performed. RESULTS: Both patients presented with fairly sudden onset of profound and persistent memory loss in the absence of other neurological symptoms. Their subsequently diagnosed small cell lung cancer was treated with a combination of radiotherapy and chemotherapy, leading to remission of the tumour. The memory of patient 1 recovered fully and he died from an unrelated cause 1 year later; neuropsychological testing showed a severe, but isolated, anterograde amnesia, brain MRI was normal and HMPAO-SPECT showed left medial temporal hypoperfusion. Patient 2 remained densely amnesic despite regression of her lung tumour; neuropsychological testing disclosed both anterograde and extensive retrograde amnesia together with more generalised cognitive deficits including anomia and executive impairments, MRI showed gross atrophy of the hippocampus and amygdala bilaterally, and HMPAO-SPECT showed pronounced frontal and temporal hypoperfusion. CONCLUSION: Complete remission from PNLE may occur and seems to be associated with pure anterograde amnesia without evidence of structural hippocampal damage in MRI. By contrast, cognitive deficits beyond severe anterograde amnesia and evidence of destructive medial temporal lobe pathology on MRI seem to be poor prognostic features. PMID- 11413261 TI - Interference between postural control and mental task performance in patients with vestibular disorder and healthy controls. AB - OBJECTIVES: To determine whether interference between postural control and mental task performance in patients with balance system impairment and healthy subjects is due to general capacity limitations, motor control interference, competition for spatial processing resources, or a combination of these. METHOD: Postural stability was assessed in 48 patients with vestibular disorder and 24 healthy controls while they were standing with eyes closed on (a) a stable and (b) a moving platform. Mental task performance was measured by accuracy and reaction time on mental tasks, comprising high and low load, spatial and non-spatial tasks. Interference between balancing and performing mental tasks was assessed by comparing baseline (single task) levels of sway and mental task performance with levels while concurrently balancing and carrying out mental tasks. RESULTS: As the balancing task increased in difficulty, reaction times on both low load mental tasks grew progressively longer and accuracy on both high load tasks declined in patients and controls. Postural sway was essentially unaffected by mental activity in patients and controls. CONCLUSIONS: It is unlikely that dual task interference between balancing and mental activity is due to competition for spatial processing resources, as levels of interference were similar in patients with vestibular disorder and healthy controls, and were also similar for spatial and non-spatial tasks. Moreover, the finding that accuracy declined on the high load tasks when balancing cannot be attributed to motor control interference, as no motor control processing is involved in maintaining accuracy of responses. Therefore, interference between mental activity and postural control can be attributed principally to general capacity limitations, and is hence proportional to the attentional demands of both tasks. PMID- 11413262 TI - Vestibular function in severe bilateral vestibulopathy. AB - OBJECTIVES: To assess residual vestibular function in patients with severe bilateral vestibulopathy comparing low frequency sinusoidal rotation with the novel technique of random, high acceleration rotation of the whole body. METHODS: Eye movements were recorded by electro-oculography in darkness during passive, whole body sinusoidal yaw rotations at frequencies between 0.05 and 1.6 Hz in four patients who had absent caloric vestibular responses. These were compared with recordings using magnetic search coils during the first 100 ms after onset of whole body yaw rotation at peak accelerations of 2800 degrees /s(2). Off centre rotations added novel information about otolithic function. RESULTS: Sinusoidal yaw rotations at 0.05 Hz, peak velocity 240 degrees/s yielded minimal responses, with gain (eye velocity/head velocity)<0.02, but gain increased and phase decreased at frequencies between 0.2 and 1.6 Hz in a manner resembling the vestibulo-ocular reflex. By contrast, the patients had profoundly attenuated responses to both centred and eccentric high acceleration transients, representing virtually absent responses to this powerful vestibular stimulus. CONCLUSION: The analysis of the early ocular response to random, high acceleration rotation of the whole body disclosed a profound deficit of semicircular canal and otolith function in patients for whom higher frequency sinusoidal testing was only modestly abnormal. This suggests that the high frequency responses during sinusoidal rotation were of extravestibular origin. Contributions from the somatosensory or central predictor mechanisms, might account for the generation of these responses. Random, transient rotation is better suited than steady state rotation for quantifying vestibular function in vestibulopathic patients. PMID- 11413263 TI - Magnetic resonance markers of ischaemia: their correlation with vasodilatory reserve in patients with carotid artery stenosis and occlusion. AB - OBJECTIVES: Better methods of identifying patients with asymptomatic carotid artery stenosis who are at high risk of stroke are required. It has been suggested that proton magnetic resonance spectroscopy (MRS) may allow the identification of ongoing ischaemia in this patient group by the detection of a potentially reversible reduction of N-acetyl aspartate (NAA), a presumed marker of neuronal integrity, and the presence of lactate, a marker of anaerobic metabolism. Previous studies have reported metabolite ratios rather than absolute concentrations. This study was performed to determine if NAA was reduced ipsilateral to carotid stenosis or occlusion, and if its concentration was related to carbon dioxide reactivity, a marker of cerebrovascular reserve. METHODS: Twenty one patients with unilateral carotid stenosis (>70%) or occlusion were studied. Single voxel proton MRS was performed in the ipsilateral and contralateral hemispheres, with the voxel positioned in the arterial borderzone region between the middle and anterior cerebral artery territories. Absolute quantification of metabolite concentrations was performed. Cerebrovascular reactivity to 6% carbon dioxide was determined in both middle cerebral artery territories using transcranial Doppler ultrasonography. RESULTS: Mean (SD) cerebrovascular reactivity was significantly lower in the stenosed compared with the contralateral hemisphere (13.3 (7.7) v 19.2 (8.2)%/kPa, p=0.002). There were no significant differences in the absolute concentrations of NAA, choline, or creatine between the ipsilateral and contralateral hemispheres (for example, NAA 10.1 (1.1) v 10.5 (1.1) mmol/l, p=0.1). No lactate peak was seen in any spectra. For each metabolite measured, there was no correlation between the absolute concentration and cerebrovascular reactivity for either hemisphere. CONCLUSIONS: In patients with carotid stenosis and occlusion we found no evidence that chronic hypoperfusion is associated with a reduction in NAA or the presence of lactate. Magnetic resonance spectroscopy is unlikely to help in the selection of patients with asymptomatic carotid stenosis for endarterectomy. PMID- 11413264 TI - Emil Adolph von Behring (1854-1917) and Shibasaburo Kitasato (1852-1931). PMID- 11413265 TI - Randomised controlled trial of telemedicine for new neurological outpatient referrals. AB - OBJECTIVE: To test the hypothesis that telemedicine for new patient referrals to neurological outpatients is as efficient and acceptable as conventional face to face consultation. METHODS: A randomised controlled trial between two groups: face to face (FF) and telemedicine (TM). This study was carried out between a neurological centre and outlying clinics at two distant hospitals linked by identical medium cost commercial interactive video conferencing equipment with ISDN lines transmitting information at 384 kbits/s. The same two neurologists carried out both arms of the study. Of the 168 patients who were suitable for the study, 86 were randomised into the telemedicine group and 82 into the face to face group. Outcome measures were (1) consultation process: (a) number of investigations; (b) number of drugs prescribed; (c) number of patient reviews and (2) patient satisfaction: (a) confidence in consultation; (b) technical aspects of consultation; (c) aspects surrounding confidentiality. Diagnostic categories were also measured to check equivalence between the groups: these were structural neurological, structural non-neurological, non-structural, and uncertain. RESULTS: Diagnostic categories were similar (p>0.5) between the two groups. Patients in the telemedicine group had significantly more investigations (p=0.001). There was no difference in the number of drugs prescribed (p>0.5). Patients were generally satisfied with both types of consultation process except for concerns about confidentiality and embarrassment in the telemedicine group (p=0.017 and p=0.005 respectively). CONCLUSION: Telemedicine for new neurological outpatients is possible and feasible but generates more investigations and is less well accepted than face to face examination. PMID- 11413266 TI - Prospective study of swallowing function in patients with cervical dystonia undergoing selective peripheral denervation. AB - OBJECTIVE: To characterise swallowing function in patients with cervical dystonia with botulinum toxin treatment failure, before and after selective peripheral denervation surgery. METHODS: Twelve patients with cervical dystonia had a thorough examination including standardised assessment for cervical dystonia, scoring of subjective dysphagia, and videofluoroscopic swallow. Videofluoroscopy was scored by consensus opinion between a speech and language therapist and an independent blinded radiologist using a validated scoring system. RESULTS: Seven patients with cervical dystonia experienced no subjective dysphagia either before or after surgery, although in all these patients there was objective videofluoroscopic evidence of underlying mild to moderate oropharyngeal dysphagia preoperatively and postoperatively. The most common finding was delayed initiation of swallow. Three other patients, also without subjective dysphagia before surgery, developed postoperative dysphagia. In these patients, videofluoroscopy showed a delayed swallow reflex before surgery, which was worse postoperatively in two. The remaining two patients had mild subjective dysphagia before surgery that improved postoperatively in one and deteriorated in the other. In the first, videofluoroscopy was normal preoperatively and postoperatively, and in the second, oral bolus preparation was moderately abnormal preoperatively and swallow initiation was delayed postoperatively. Mean subjective dysphagia scores did not change significantly. Apart from a significant improvement of tongue base retraction, videofluoroscopic scores were not significantly different after surgery. Postoperatively there was significant improvement of overall cervical dystonia severity and abnormal head rotation in the group as a whole. There was no correlation between age, duration of symptoms of cervical dystonia, preoperative or postoperative cervical dystonia severity, subjective dysphagia scores, or videofluoroscopic scores. However, in the five patients with persisting anterior sagittal head shift as part of the torticollis, tongue base retraction was less likely to improve after surgery compared with those without head shift. CONCLUSION: Surgical denervation of dystonic neck muscles, leading to improved neck posture, can also improve tongue base retraction, which is a key component of normal bolus propagation. However, delayed swallow initiation, a common feature in patients with cervical dystonia, can be further compromised by surgery, leading to subjective dysphagia. In general, selective peripheral denervation seems to be a safe procedure with no major compromise of swallowing function. PMID- 11413267 TI - Prospective study of omental transposition in patients with chronic spinal injury. AB - OBJECTIVES: This prospective study was designed to assess the effects of omental transposition in patients with a chronic spinal injury. METHODS: Neurological status was established to be stable and multiple baseline across patient studies were done preoperatively and repeated postoperatively. Assessments included activities of daily living (ADL), functional ability, degree of spasticity, motor power, sensation, pain perception, urodynamic studies, electromyography, sensory evoked potentials (SEPs), and infrared thermography to measure peripheral and general skin vascular responses. Each patient had MRI. Assessments were done at 3, 6, and 12 months after omental transposition in 17 patients. RESULTS: The detailed assessments failed to show significant improvement, although some patients showed minor objective and subjective change in some categories. Neurological deterioration occurred in one patient. There were 20 surgical complications including urinary tract infection, deep vein thrombosis, wound infection, and incisional hernia. CONCLUSIONS: Omental transposition has not been shown to improve neurological function in 17 patients with chronic spinal cord injury, and continued use of this operation in this situation is not supported by this study. Further advances in spinal cord repair may utilise the pedicled omental graft to provide an alternative vascular supply, but its current use should be limited to experimental models. PMID- 11413268 TI - Ataxia associated with Hashimoto's disease: progressive non-familial adult onset cerebellar degeneration with autoimmune thyroiditis. AB - Acquired cerebellar ataxia has been described with hypothyroidism, and is typically reversible by thyroid hormone replacement therapy. The cerebellar dysfunction has been attributed to metabolic and physiological effects of the endocrine disorder. In a few patients, however, ataxia has persisted despite thyroid replacement therapy. Other mechanisms may be involved in ataxia associated with thyroid disorders. OBJECTIVE: To document progressive non familial adult onset cerebellar degeneration (PNACD) occurring in six patients with raised antithyroid antibodies (Hashimoto's/autoimmune thyroiditis), and other autoimmune manifestations, in the absence of hypothyroidism; and to document the independence of the cerebellar disorder from the endocrine dysfunction. METHODS: A case study of six patients with PNACD reviewing the clinical course and relation to endocrine and autoimmune status. RESULTS: All six patients were euthyroid when they developed their symptoms; had raised antithyroid antibodies consistent with Hashimoto's autoimmune thyroiditis; and had strong personal or family histories of organ specific autoimmune diatheses. Brain MRI disclosed atrophy of the cerebellar vermis in four patients and olivopontocerebellar atrophy in two. Other possible causes of cerebellar degeneration were excluded. De novo treatment (two patients) or continued treatment (three patients) with L-thyroxine did not modify the progression of the ataxia. CONCLUSIONS: Cerebellar degeneration in these patients with raised antithyroid antibodies may be immune mediated. The presence of antithyroid antibodies may signal or cause the autoimmune process producing cerebellar degeneration. "Hashimoto's associated ataxia" seems to represent a recognisable and not uncommon condition; a trial of immunomodulating therapy should be considered in these patients. PMID- 11413269 TI - Polyamine metabolism in brain tumours: diagnostic relevance of quantitative biochemistry. AB - OBJECTIVE: Activation of polyamine metabolism is closely associated with cellular proliferation. The purpose was to investigate whether the content of the polyamines putrescine, spermidine, and spermine, and the activity of the first metabolic key enzyme of polyamine metabolism, ornithine decarboxylase (ODC), represent biochemical markers of malignancy in brain tumours. METHODS: The concentration of putrescine, spermidine, and spermine, and the activity of ODC were biochemically quantified in tissue samples obtained during open microsurgery of 670 patients with brain tumours. Biochemical analysis and histopathological classification were carried out in serial tumour samples. RESULTS: The activity of ODC was very low in peritumoral non-neoplastic brain tissue (0.9 (SD 0.6) nmol/g/h). It was significantly higher in gliomas and it significantly increased with a higher grade of malignancy (grade I 2.7 (2.8) nmol/g/h, grade II 3.1 (4.0) nmol/g/h, grade III 5.7 (5.6) nmol/g/h, grade IV 10.6 (11.7) nmol/g/h). High enzyme activity was also found in medulloblastomas (25.5 (15.1) nmol/g/h), malignant lymphomas (52.1 (42.1) nmol/g/h), and metastases from carcinoma (14.9 (22.1) nmol/g/h). Lowest values were measured in epidermoid cysts (0.5 (0.2) nmol/g/h), craniopharyngiomas (1.2 (0.9) nmol/g/h), angioblastomas (1.6 (1.7) nmol/g/h), and neurinomas (2.0 (1.8) nmol/g/h). By contrast with ODC activity, polyamine concentrations did not correlate with the grade of malignancy. Correlation of regional biochemical and histomorphological data in rapidly growing neoplasms showed high enzyme activity in solid tumour parts and low activity in necrotic areas. CONCLUSIONS: Novel data relating ODC activation and polyamine concentrations to neuropathology is presented indicating that high ODC activity represents a biochemical marker of malignancy in brain tumours. This information is important for clinical and therapeutic investigations. PMID- 11413270 TI - Long term effects of locomotor training in spinal humans. AB - The long term effects of locomotor training in patients with spinal cord injury (SCI) were studied. In patients with complete or incomplete SCI coordinated stepping movements were induced and trained by bodyweight support and standing on a moving treadmill. The leg extensor muscle EMG activity in both groups of patients increased significantly over the training period, associated with improved locomotor ability in those with incomplete SCI. During a period of more than 3 years after training, the level of leg extensor EMG remained about constant in incomplete SCI in those who regularly maintained locomotor activity. By contrast the EMG significantly fell in those with complete SCI. The results suggest a training induced plasticity of neuronal centres in the isolated spinal cord which may be of relevance for future interventional therapies. PMID- 11413271 TI - No association between multiple sclerosis and the Notch3 gene responsible for cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). AB - The clinical and radiological overlap between multiple sclerosis and cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL; MIM 125310) raises the possibility of diagnostic confusion and suggests that pleiotropic effects of the Notch3 gene might include influencing susceptibility to multiple sclerosis. To investigate these possibilities three microsatellites markers closely flanking the Notch 3 gene in 745 simplex families with multiple sclerosis were genotyped and exon 3 and exon 4 of the gene were directly sequenced in a subset of the index members from these families (n=93). No evidence for association was found in any of the three markers and none of the commoner mutations causing CADASIL were found in the sequenced patients. PMID- 11413272 TI - Antibodies against glutamic acid decarboxylase: prevalence in neurological diseases. AB - High prevalence of autoantibodies against glutamic acid decarboxylase (GAD-Ab) in stiff man syndrome (SMS) not only helps diagnosis, but also suggests immune mediated impairment of GABAergic functions. However, the presence of GAD-Ab has also been reported in other neurological syndromes. Therefore the prevalence of GAD-Ab was investigated in SMS, progressive encephalomyelitis with rigidity and myoclonus (PERM), and in other neurological diseases (OND). Serum antibodies against the GAD isoforms, GAD65 and GAD67, were investigated with radioimmunoassays in 13 patients with SMS, nine with PERM, 279 consecutive patients with OND, and in 100 normal controls. RESULTS: Prevalence of GAD65Ab was around 80% in patients with SMS/PERM compared with 5% in patients with OND and 1% in normal controls. Prevalence of GAD67Ab was 60% in SMS/PERM, 2% in patients with OND, and 1% in normal controls. Raised GAD-Ab clustered in an OND subgroup with sporadic progressive ataxia, but not in OND subgroups with recognised neuroimmunological diseases. In conclusion, increased GAD-Ab is neither a non specific epiphenomenon of neuronal damage nor a common feature of recognised neuroimmunological disorders. In neurological diseases, GAD-Ab may be a pathogenetic agent or a marker for an ongoing autoimmune process, or both. PMID- 11413273 TI - Alcohol consumption and frontal lobe shrinkage: study of 1432 non-alcoholic subjects. AB - OBJECTIVES: To evaluate the influences of chronic alcohol consumption on brain volume among social drinkers, as it is well known that alcohol misusers have a high risk of brain shrinkage. METHODS: Frontal lobe volumes on MRI were compared with the current alcohol habits of consecutive 1432 non-alcoholic subjects. RESULTS: After adjusting for other variables, age was found to be the most powerful promoting factor for the shrinkage with a odds ratio of 2.8 (95% confidence interval (95% CI) 1.23-3.06) for each 10 years of age. Regarding alcohol habit, 667 of the subjects were abstainers, and 157, 362, and 246 of the subjects were light (average 88.2 g ethanol/week), moderate (181.2 g/week), and heavy (418.1 g/week) drinkers, respectively. Moderate alcohol consumption did not increase the incidence of frontal lobe shrinkage (odds ratio 0.98; 95% CI 0.73 1.33), whereas heavy drinkers were at a higher risk compared with abstainers (1.80; 1.32-2.46). The contributory rate of alcohol consumption for frontal lobe shrinkage was 11.3%. CONCLUSION: The brain tends to shrink physiologically with age. Heavy alcohol consumption seems to exaggerate this shrinkage in social drinkers. Moderate alcohol consumption does not seem to affect brain volume. PMID- 11413274 TI - Gabapentin but not vigabatrin is effective in the treatment of acquired nystagmus in multiple sclerosis: How valid is the GABAergic hypothesis? AB - Acquired nystagmus occurs frequently in patients with multiple sclerosis and is often the cause of illusory motion of the environment (oscillopsia), and blurring of vision. Based primarily on the beneficial effect of gabapentin on acquired pendular nystagmus (APN), a GABAergic mechanism in controlling nystagmus has been hypothesised. If increasing GABA concentrations in the CNS are critical for the treatment of nystagmus, then a selective GABAergic drug should be highly successful. However, as gabapentin is not a selective GABAergic agent, vigabatrin, a "pure" GABAergic medication, and gabapentin, were compared in a single blind cross over trial in eight patients with definite multiple sclerosis. Patients were randomly assigned to begin with gabapentin (1200 mg daily) or vigabatrin (2000 mg daily). Neuro-ophthalmological and electro-oculographic (EOG) evaluations were performed four and three times, respectively. Treatment efficacy was based on improving visual acuity and EOG indices (amplitude or frequency of nystagmus, or both) by at least 50% of pretreatment values. Three out of eight patients dropped out due to adverse effects. In the remaining five patients gabapentin improved symptomatic pendular or gaze evoked jerk nystagmus in four. Three patients decided to continue gabapentin therapy. Importantly, vigabatrin proved useful in only one out of five patients, suggesting that gabapentin effectiveness may be related to additional non-GABAergic mechanisms of action. Interaction with cerebral glutamate transmission by inhibition of NMDA receptor might be an alternative hypothesis for the therapeutic action of gabapentin. PMID- 11413275 TI - Biochemical alterations during medication withdrawal in Parkinson's disease with and without neuroleptic malignant-like syndrome. AB - The object was to assess alterations in CSF concentrations of monoamine metabolites during withdrawal of medication in patients with Parkinson's disease in relation to the presence or absence of episodes resembling neuroleptic malignant syndrome (NMS). This syndrome is a fatal condition developing after neuroleptic therapy, and a neuroleptic malignant-like syndrome (NMLS) may also occur after withdrawal of antiparkinsonian drugs in patients with Parkinson's disease. Previous biochemical assays showed that the CSF concentration of the dopamine metabolite homovanillic acid (HVA) is an independent prognostic factor for development of NMLS in patients with Parkinson's disease. In the present study, CSF concentrations of HVA, the noradrenaline (norepinephrine) metabolite 3 methoxy-4-hydroxyphenylethylene glycol, and the serotonin metabolite 5 hydroxyindole acetic acid were assayed using high performance liquid chromatography with electrochemical detection. The study population consisted of nine patients with Parkinson's disease with NMLS and 12 without NMLS, in whom metabolites were assayed during both withdrawal and remedicated periods. Concentrations of HVA in the CSF were significantly lower during the withdrawal period than the medicated period regardless of whether patients developed NMLS, and HVA concentrations were comparably increased after remedication in both groups. However, HVA concentrations were significantly lower in patients with NMLS than in those without NMLS during both withdrawal and medicated periods. Other metabolites showed no significant differences. The present data provide further biochemical evidence for extremely suppressed central dopaminergic activity during NMLS, which may indicate a narrow safety margin for medication withdrawal in patients with Parkinson's disease. PMID- 11413276 TI - Right side neglect in right cerebellar lesion. AB - A patient is described who developed right side hemineglect after a right cerebellar lesion. This spatial disorder was interpreted as a secondary effect of a deficit of the motor organisation in the right hemispace due to left frontal diaschisis. The pathological base may be the interruption of a highly integrated system which includes the lateral cerebellum and the contralateral frontal lobe. PMID- 11413277 TI - Combined systemic and intraventricular chemotherapy in primary CNS lymphoma: a pilot study. AB - The objective was to evaluate response rate, response duration, and toxicity after systemic and intraventricular chemotherapy in primary CNS lymphoma (PCNSL). From September 1995 to September 1998, 20 consecutive patients with PCNSL (median age 64, range 27 to 71 years) were enrolled in a pilot study evaluating chemotherapy without radiotherapy. A high dose methotrexate (MTX) (cycles 1, 2, 4, 5) and cytarabine (ara-C) (cycles 3, 6) based systemic therapy (including dexamethasone, vinca alkaloids, ifosfamide, and cyclophosphamide) was combined with intraventricular MTX, prednisolone, and ara-C. Complete response was achieved in 11 and partial remission in two patients; in one response could not be determined. Four patients showed progressive disease and two (70, 71 years) died from treatment related complications. Observation time was 2 to 59 months (median 31.5 months). Kaplan-Meier estimate for median time to treatment failure (TTF) was 20.5 months, and for median survival 54 months. Systemic toxicity was mainly hematological. Ommaya reservoir infection occurred in four patients and acute transient MTX induced encephalopathy in two (subacute in another). Cognitive dysfunction possibly due to treatment was seen in only one patient after relapse and after a total of 12 cycles (six at relapse). In conclusion, primary chemotherapy based on high dose MTX and ara-C is highly efficient in PCNSL. Toxicity is manageable in patients younger than 70 years. PMID- 11413278 TI - Range of cross reactivity of anti-GM1 IgG antibody in Guillain-Barre syndrome. AB - The cross reactivity of anti-GM1 IgG antibody with various gangliosides and asialo-GM1 in serum samples from 27 patients with Guillain-Barre syndrome was investigated. An enzyme linked immunosorbent assay (ELISA) absorption study showed that anti-GM1 IgG antibody cross reacted with asialo-GM1 in 52% of the patients, GM1b in 41%, GD1b in 22%, and GalNAc-GD1a in 19%, and that it did not cross react with GM2, GT1b, or GQ1b. The antibody that cross reacted with GD1b was associated with a high frequency of cranial nerve involvement and negative Campylobacter jejuni serology. Anti-GM1 IgG antibody has a broad range of cross reactivity which may contribute to various clinical variations of Guillain-Barre syndrome. PMID- 11413279 TI - Brain stem compression by a giant vertebrobasilar aneurysm mimicking seronegative myasthenia. AB - A patient is described with a vertebrobasilar aneurysm who was erroneously thought to have myasthenia gravis on the basis of the clinical presentation and investigations, which were interpreted as supportive of a disorder of the neuromuscular junction. Despite the correct diagnosis being made at a late stage the patient made a full recovery after radiological intervention. PMID- 11413280 TI - Spontaneous occipital subdural haematoma. PMID- 11413281 TI - Peripheral axonal motor degeneration after spinal cord infarct. PMID- 11413291 TI - RNA recombination between persisting pestivirus and a vaccine strain: generation of cytopathogenic virus and induction of lethal disease. AB - Molecular analysis of a cytopathogenic (cp) bovine viral diarrhea virus (BVDV) isolate (1741) obtained from a case of mucosal disease (MD) led to the identification of five different viral subgenomic RNAs in addition to a noncytopathogenic (noncp) strain (NCP 1741). For each of the subgenomes, a large internal deletion was found together with an inserted sequence encoding part of ribosomal protein S27a fused to an N-terminally truncated ubiquitin monomer. Surprisingly, the two cellular insertions together with flanking viral sequences encoding parts of NS3 and NS4B are >99% identical to the previously described sequence of BVDV vaccine strain RIT (P. Becher, M. Orlich, and H.-J. Thiel, J. Virol. 72:8697-8704, 1998), while the remainder of the subgenomes is derived from the genome of NCP 1741. Further analyses including molecular cloning and nucleotide sequencing of the recombination partners revealed that both homologous and nonhomologous RNA recombination contributed to the generation of the viral subgenomes. Interestingly, for another cp BVDV isolate (CP 4584) from an independent case of MD, again an insertion of a RIT-derived sequence element was detected. In contrast to CP 1741, for CP 4584 a duplication of the genomic region encoding NS3 and parts of NS4A and NS4B was found. Transfection of bovine cells with RNA transcribed from a chimeric cDNA construct showed that the RIT-derived insertion together with the CP 4584-specific duplication of viral sequences represents the genetic basis of cytopathogenicity of CP 4584. Remarkably, passages of the recovered cp virus in cell culture led to emergence of noncp BVDV and a number of viral subgenomes whose genome organization was similar to that in BVDV 1741. PMID- 11413290 TI - Virus-specific mRNA capping enzyme encoded by hepatitis E virus. AB - Hepatitis E virus (HEV), a positive-strand RNA virus, is an important causative agent of waterborne hepatitis. Expression of cDNA (encoding amino acids 1 to 979 of HEV nonstructural open reading frame 1) in insect cells resulted in synthesis of a 110-kDa protein (P110), a fraction of which was proteolytically processed to an 80-kDa protein. P110 was tightly bound to cytoplasmic membranes, from which it could be released by detergents. Immunopurified P110 catalyzed transfer of a methyl group from S-adenosylmethionine (AdoMet) to GTP and GDP to yield m(7)GTP or m(7)GDP. GMP, GpppG, and GpppA were poor substrates for the P110 methyltransferase. There was no evidence for further methylation of m(7)GTP when it was used as a substrate for the methyltransferase. P110 was also a guanylyltransferase, which formed a covalent complex, P110-m(7)GMP, in the presence of AdoMet and GTP, because radioactivity from both [alpha-(32)P]GTP and [(3)H-methyl]AdoMet was found in the covalent guanylate complex. Since both methyltransferase and guanylyltransferase reactions are strictly virus specific, they should offer optimal targets for development of antiviral drugs. Cap analogs such as m(7)GTP, m(7)GDP, et(2)m(7)GMP, and m(2)et(7)GMP inhibited the methyltransferase reaction. HEV P110 capping enzyme has similar properties to the methyltransferase and guanylyltransferase of alphavirus nsP1, tobacco mosaic virus P126, brome mosaic virus replicase protein 1a, and bamboo mosaic virus (a potexvirus) nonstructural protein, indicating there is a common evolutionary origin of these distantly related plant and animal virus families. PMID- 11413292 TI - Fidelity of leader and trailer sequence usage by the respiratory syncytial virus and avian pneumovirus replication complexes. AB - The specificity of usage of promoters for replication and transcription by the pneumoviruses human respiratory syncytial virus (HRSV) and avian pneumovirus (APV) was studied using minigenomes containing a reporter gene. When infectious HRSV or APV was used as helper virus, replication could occur only if both the leader and trailer regions (containing the replicative and transcriptional promoters) were derived from the helper virus. In contrast, when the HRSV replication complex was supplied from cDNA plasmids, a minigenome containing either the APV leader or trailer was recognized and substantial levels of replication and transcription occurred. These data suggest that in pneumovirus infected cells, helper virus functions can discriminate between genomes on the basis of the terminal sequences and that there is an association between the leader and trailer required for productive replication. This association is required only in virus-infected cells, not when replication and transcription are mediated by plasmid-directed expression of the component proteins required for replication and transcription. The possible implications of this are discussed. PMID- 11413293 TI - Common antiviral cytotoxic t-lymphocyte epitope for diverse arenaviruses. AB - Members of the Arenaviridae family have been isolated from mammalian hosts in disparate geographic locations, leading to their grouping as Old World types (i.e., lymphocytic choriomeningitis virus [LCMV], Lassa fever virus [LFV], Mopeia virus, and Mobala virus) and New World types (i.e., Junin, Machupo, Tacaribe, and Sabia viruses) (C. J. Peters, M. J. Buchmeier, P. E. Rollin, and T. G. Ksiazek, p. 1521-1551, in B. N. Fields, D. M. Knipe, and P. M. Howley [ed.], Fields virology, 3rd ed., 1996; P. J. Southern, p. 1505-1519, in B. N. Fields, D. M. Knipe, and P. M. Howley [ed.], Fields virology, 3rd ed., 1996). Several types in both groups-LFV, Junin, Machupo, and Sabia viruses-cause severe and often lethal human diseases. By sequence comparison, we noted that eight Old World and New World arenaviruses share several amino acids with the nucleoprotein (NP) that consists of amino acids (aa) 118 to 126 (NP 118-126) (RPQASGVYM) of LCMV that comprise the immunodominant cytotoxic T-lymphocyte (CTL) epitope for H-2(d) mice (32). This L(d)-restricted epitope constituted >97% of the total bulk CTLs produced in the specific antiviral or clonal responses of H-2(d) BALB mice. NP 118-126 of the Old World arenaviruses LFV, Mopeia virus, and LCMV and the New World arenavirus Sabia virus bound at high affinity to L(d). The primary H-2(d) CTL anti-LCMV response as well as that of a CTL clone responsive to LCMV NP 118 126 recognized target cells coated with NP 118-126 peptides derived from LCMV, LFV, and Mopeia virus but not Sabia virus, indicating that a common functional NP epitope exists among Old World arenaviruses. Use of site-specific amino acid exchanges in the NP CTL epitope among these arenaviruses identified amino acids involved in major histocompatibility complex binding and CTL recognition. PMID- 11413294 TI - Relative dominance of epitope-specific cytotoxic T-lymphocyte responses in human immunodeficiency virus type 1-infected persons with shared HLA alleles. AB - Cytotoxic T lymphocytes (CTL) target multiple epitopes in human immunodeficiency virus (HIV)-infected persons, and are thought to influence the viral set point. The extent to which HLA class I allele expression predicts the epitopes targeted has not been determined, nor have the relative contributions of responses restricted by different class I alleles within a given individual. In this study, we performed a detailed analysis of the CTL response to optimally defined CTL epitopes restricted by HLA class I A and B alleles in individuals who coexpressed HLA A2, A3, and B7. The eight HIV-1-infected subjects studied included two subjects with acute HIV infection, five subjects with chronic HIV infection, and one long-term nonprogressor. Responses were heterogeneous with respect to breadth and magnitude of CTL responses in individuals of the same HLA type. Of the 27 tested epitopes that are presented by A2, A3, and B7, 25 were targeted by at least one person. However, there was wide variation in the number of epitopes targeted, ranging from 2 to 17. The A2-restricted CTL response, which has been most extensively studied in infected persons, was found to be narrowly directed in most individuals, and in no cases was it the dominant contributor to the total HIV-1-specific CTL response. These results indicate that HLA type alone does not predict CTL responses and that numerous potential epitopes may not be targeted by CTL in a given individual. These data also provide a rationale for boosting both the breadth and the magnitude of HIV-1-specific CTL responses by immunotherapy in persons with chronic HIV-1 infection. PMID- 11413295 TI - Products of US22 genes M140 and M141 confer efficient replication of murine cytomegalovirus in macrophages and spleen. AB - Efficient replication of murine cytomegalovirus (MCMV) in macrophages is a prerequisite for optimal growth and spread of the virus in its natural host. Simultaneous deletion of US22 gene family members M139, M140, and M141 results in impaired replication of MCMV in macrophages and mice. In this study, we characterized the proteins derived from these three genes and examined the impact of individual gene deletions on viral pathogenesis. The M139, M140, and M141 gene products were identified as early proteins that localize to both the nucleus and cytoplasm in infected cells. Gene M139 encodes two proteins, of 72 and 61 kDa, while M140 and M141 each encode a single protein of 56 (pM140) and 52 (pM141) kDa, respectively. No role for the M139 proteins in MCMV replication in macrophages or mice was determined in these studies. In contrast, deletion of either M140 or M141 resulted in impaired MCMV replication in macrophages and spleen tissue. Replication of the M140 deletion mutant was significantly more impaired than that of the virus lacking M141. Further analyses revealed that the absence of the pM140 adversely affected pM141 levels by rendering the latter protein unstable. Since the replication defect due to deletion of M140 was more profound than could be explained by the reduced half-life of pM141, pM140 must exert an additional, independent function in mediating efficient replication of MCMV in macrophages and spleen tissue. These data indicate that the US22 genes M140 and M141 function both cooperatively and independently to regulate MCMV replication in a cell type-specific manner and, thus, to influence viral pathogenesis. PMID- 11413296 TI - An amino acid substitution in the coding region of the E2 glycoprotein adapts Ross River virus to utilize heparan sulfate as an attachment moiety. AB - Passage of Ross River virus strain NB5092 in avian cells has been previously shown to select for virus variants that have enhanced replication in these cells. Sequencing of these variants identified two independent sites that might be responsible for the phenotype. We now demonstrate, using a molecular cDNA clone of the wild-type T48 strain, that an amino acid substitution at residue 218 in the E2 glycoprotein can account for the phenotype. Substitutions that replaced the wild-type asparagine with basic residues had enhanced replication in avian cells while acidic or neutral residues had little or no observable effect. Ross River virus mutants that had increased replication in avian cells also grew better in BHK cells than the wild-type virus, whereas the remaining mutants were unaffected in growth. Replication in both BHK and avian cells of Ross River virus mutants N218K and N218R was inhibited by the presence of heparin or by the pretreatment of the cells with heparinase. Binding of the mutants, but not of the wild type, to a heparin-Sepharose column produced binding comparable to that of Sindbis virus, which has previously been shown to bind heparin. Replication of these mutants was also adversely affected when they were grown in a CHO cell line that was deficient in heparan sulfate production. These results demonstrate that amino acid 218 of the E2 glycoprotein can be modified to create an heparan sulfate binding site and this modification expands the host range of Ross River virus in cultured cells to cells of avian origin. PMID- 11413297 TI - A single amino acid alteration in the human parainfluenza virus type 3 hemagglutinin-neuraminidase glycoprotein confers resistance to the inhibitory effects of zanamivir on receptor binding and neuraminidase activity. AB - Entry and fusion of human parainfluenza virus type 3 (HPF3) requires interaction of the viral hemagglutinin-neuraminidase (HN) glycoprotein with its sialic acid receptor. 4-Guanidino-2,4-dideoxy-2,3-dehydro-N-acetylneuraminic acid (4-GU-DANA; zanamivir), a sialic acid transition-state analog designed to fit the influenza virus neuraminidase catalytic site, possesses antiviral activity at nanomolar concentrations in vitro. We have shown previously that 4-GU-DANA also inhibits both HN-mediated binding of HPF3 to host cell receptors and HN's neuraminidase activity. In the present study, a 4-GU-DANA-resistant HPF3 virus variant (ZM1) was generated by serial passage in the presence of 4-GU-DANA. ZM1 exhibited a markedly fusogenic plaque morphology and harbored two HN gene mutations resulting in two amino acid alterations, T193I and I567V. Another HPF3 variant studied in parallel, C-0, shared an alteration at T193 and exhibited similar plaque morphology but was not resistant to 4-GU-DANA. Neuraminidase assays revealed a 15 fold reduction in 4-GU-DANA sensitivity for ZM1 relative to the wild type (WT) and C-0. The ability of ZM1 to bind sialic acid receptors was inhibited 10-fold less than for both WT and C-0 in the presence of 1 mM 4-GU-DANA. ZM1 also retained infectivity at 15-fold-higher concentrations of 4-GU-DANA than WT and C 0. A single amino acid alteration at HN residue 567 confers these 4-GU-DANA resistant properties. An understanding of ZM1 and other escape variants provides insight into the effects of this small molecule on HN function as well as the role of the HN glycoprotein in HPF3 pathogenesis. PMID- 11413298 TI - YADD mutants of human immunodeficiency virus type 1 and Moloney murine leukemia virus reverse transcriptase are resistant to lamivudine triphosphate (3TCTP) in vitro. AB - When human immunodeficiency virus type 1 (HIV-1) is selected for resistance to 3TC, the methionine normally present at position 184 is replaced by valine or isoleucine. Position 184 is the X of the conserved YXDD motif; positions 185 and 186 form part of the triad of aspartic acids at the polymerase active site. Structural and biochemical analysis of 3TC-resistant HIV-1 reverse transcriptase (RT) led to a model in which a beta-branched amino acid at position 184 would act as a steric gate. Normal deoxynucleoside triphosphates (dNTPs) could still be incorporated; the oxathiolane ring of 3TCTP would clash with the beta branch of the amino acid at position 184. This model can also explain 3TC resistance in feline immunodeficiency virus and human hepatitis B virus. However, it has been reported (14) that murine leukemia viruses (MLVs) with valine (the amino acid present in the wild type), isoleucine, alanine, serine, or methionine at the X position of the YXDD motif are all resistant to 3TC. We prepared purified wild type MLV RT and mutant MLV RTs with methionine, isoleucine, and alanine at the X position. The behavior of these RTs was compared to those of wild-type HIV-1 RT and of HIV-1 RT with alanine at the X position. If alanine is present at the X position, both MLV RT and HIV-1 RT are relatively resistant to 3TCTP in vitro. However, the mutant enzymes were impaired relative to their wild-type counterparts; there appears to be steric hindrance for both 3TCTP and normal dNTPs. PMID- 11413299 TI - A nonviral peptide can replace the entire N terminus of zucchini yellow mosaic potyvirus coat protein and permits viral systemic infection. AB - Systematic deletion and peptide tagging of the amino-terminal domain (NT, ~43 amino acids) of an attenuated zucchini yellow mosaic potyvirus (ZYMV-AGII) coat protein (CP) were used to elucidate its role in viral systemic infection. Deletion mutants truncated by 8, 13, and 33 amino acid residues from the CP-NT 5' end were systemically infectious and produced symptoms similar to those of the AGII virus. Tagging these deletion mutants with either human c-Myc (Myc) or hexahistidine peptides maintained viral infectivity. Similarly, addition of these peptides to the intact AGII CP-NT did not affect viral life cycle. To determine which parts, if any, of the CP-NT are essential for viral systemic infection, a series of Myc-tagged mutants with 8 to 43 amino acids removed from the CP-NT were constructed. All Myc-tagged CP-NT deletion mutants, including those from which virtually all the viral CP-NT had been eliminated, were able to encapsidate and cause systemic infection. Furthermore, chimeric viruses with deletions of up to 33 amino acids from CP-NT produced symptoms indistinguishable from those caused by the parental AGII virus. In contrast to CP-NT Myc fusion, addition of the foot and-mouth disease virus (FMDV) immunogenic epitope to AGII CP-NT did not permit systemic infection. However, fusion of the Myc peptide to the N terminus of the FMDV peptide restored the capability of the virus to spread systemically. We have demonstrated that all CP-NT fused peptides were exposed on the virion surface, masking natural CP immunogenic determinants. Our findings demonstrate that CP-NT is not essential for ZYMV spread and that it can be replaced by an appropriate foreign peptide while maintaining systemic infectivity. PMID- 11413300 TI - A Ty1 reverse transcriptase active-site aspartate mutation blocks transposition but not polymerization. AB - Reverse transcriptases (RTs) are found in a wide variety of mobile genetic elements including viruses, retrotransposons, and infectious organellar introns. An invariant triad of aspartates is thought to be required for the catalytic function of RTs. We generated RT mutants in the yeast retrotransposon Ty1, changing each of these active-site aspartates to asparagine or glutamate. All but one of the mutants lacked detectable polymerase activity. The novel exception, D(211)N, retained near wild-type in vitro polymerase activity within virus-like particles but failed to carry out in vivo transposition. For this mutant, minus strand synthesis is impaired and formation of the plus-strand strong-stop intermediate is eliminated. Intragenic second-site suppressor mutations of the transposition defect map to the RNase H domain of the enzyme. Our results demonstrate that one of the three active-site aspartates in a retrotransposon RT is not catalytically critical. This implies a basic difference in the polymerase active-site geometry of Ty1 and human immunodeficiency virus RT and shows that subtle mutations in one domain can cause dramatic functional effects on a distant domain of the same enzyme. PMID- 11413301 TI - Evidence for retroviral intramolecular recombinations. AB - As a consequence of being diploid, retroviruses have a high recombination rate. Naturally occurring retroviruses contain two repeat sequences (R regions) flanking either end of their RNA genomes, and recombination between these two R regions occurs at a high rate. We deduced that recombination may occur between two sequences within the same RNA molecule (intramolecular) as well as between sequences present within two separate RNA molecules (intermolecular). Intramolecular recombination would usually result in a deletion within the progeny provirus. In this report, we demonstrate that intramolecular recombination between two identical sequences occurred within a chimeric RNA vector. In addition, high rates of recombination between two identical sequences within the same RNA molecule resulted mostly from intramolecular recombination. PMID- 11413302 TI - Hepatitis B virus core antigen binds and activates naive human B cells in vivo: studies with a human PBL-NOD/SCID mouse model. AB - The hepatitis B virus (HBV) core (HBc) antigen (HBcAg) is a highly immunogenic subviral particle. Studies with mice have shown that HBcAg can bind and activate B cells in a T-cell-independent fashion. By using a human peripheral blood leukocyte (hu-PBL)-Nod/LtSz-Prkdc(scid)/Prkdc(scid) (NOD/SCID) mouse model, we show here that HBcAg also activates human B cells in vivo in a T-cell-independent way. HBcAg was capable of inducing the secretion of HBcAg-binding human immunoglobulin M (IgM) in naive human B cells derived from adult human and neonatal (cord blood) donors when these hu-PBL were transferred directly into the spleens of optimally conditioned NOD/SCID mice. No such responses were found in chimeric mice that were given hu-PBL plus HBV e antigen or hu-PBL plus phosphate buffered saline. In addition, HBcAg activated purified human B cells to produce anti-HBc IgM in the chimeric mice, thus providing evidence that HBcAg behaves as a T-cell-independent antigen in humans. However, HBcAg-activated hu-PBL from naive donors were unable to switch from IgM to IgG production, even after a booster dose of HBcAg. Production of HBcAg-specific IgG could only be induced when hu-PBL from subjects who had recovered from or had an ongoing chronic HBV infection were transferred into NOD/SCID mice. Our data suggest that humans also have a population of naive B cells that can bind HBcAg and is subsequently activated to produce HBcAg-binding IgM. PMID- 11413303 TI - Molecular basis for the interaction of the hepatitis B virus core antigen with the surface immunoglobulin receptor on naive B cells. AB - The nucleocapsid of the hepatitis B virus (HBV) is composed of 180 to 240 copies of the HBV core (HBc) protein. HBc antigen (HBcAg) capsids are extremely immunogenic and can activate naive B cells by cross-linking their surface receptors. The molecular basis for the interaction between HBcAg and naive B cells is not known. The functionality of this activation was evidenced in that low concentrations of HBcAg, but not the nonparticulate homologue HBV envelope antigen (HBeAg), could prime naive B cells to produce anti-HBc in vitro with splenocytes from HBcAg- and HBeAg-specific T-cell receptor transgenic mice. The frequency of these HBcAg-binding B cells was estimated by both hybridoma techniques and flow cytometry (B7-2 induction and direct HBcAg binding) to be approximately 4 to 8% of the B cells in a naive spleen. Cloning and sequence analysis of the immunoglobulin heavy- and light-chain variable (VH and VL) domains of seven primary HBcAg-binding hybridomas revealed that six (86%) were related to the murine and human VH1 germ line gene families and one was related to the murine VH3 family. By using synthetic peptides spanning three VH1 sequences, one VH3 sequence, and one VLkappaV sequence, a linear motif in the framework region 1 (FR1)complementarity-determining region 1 (CDR1) junction of the VH1 sequence was identified that bound HBcAg. Interestingly, the HBcAg binding motif was present in the VL domain of the HBcAg-binding VH3-encoded antibody. Finally, two monoclonal antibodies containing linear HBcAg-binding motifs blocked HBcAg presentation by purified naive B cells to purified HBcAg primed CD4(+) T cells. Thus, the ability of HBcAg to bind and activate a high frequency of naive B cells seems to be mediated through a linear motif present in the FR1-CDR1 junction of the heavy or light chain of the B-cell surface receptor. PMID- 11413304 TI - Many nonmammalian cells exhibit postentry blocks to transduction by gammaretroviruses pseudotyped with various viral envelopes, including vesicular stomatitis virus G glycoprotein. AB - Previous studies have suggested that Moloney murine leukemia virus (MoMLV)-based vectors pseudotyped with the vesicular stomatitis virus G glycoprotein (VSV-G) have extensive ability to transduce nonmammalian cells. However, we have identified multiple cell lines from fish (FHM), mosquitoes (Mos-55), moths (Sf9 and High-5), flies (S2), and frogs (XPK2) that are not efficiently transduced by MoMLV-based vectors pseudotyped with many different viral envelope proteins, including VSV-G, while the same vectors are functional in these cells following transfection. A comparison of MoMLV-based vector transduction in mammalian and nonmammalian cells shows that the nonmammalian cells exhibit blocks at either entry, reverse transcription, or integration. Additionally, VSV-G-pseudotyped MoMLV-based vector transduction is attenuated in the zebrafish cell line ZF4 at entry and/or reverse transcription, whereas other transduction processes are unaffected. We show that the variation of transduction by MoMLV-based vectors in mammalian and nonmammalian cells is not due to differences in culture conditions or cell division rate but is likely the result of divergence in cellular factors required for retroviral transduction. PMID- 11413305 TI - Target cell populations of human immunodeficiency virus type 1 in peripheral blood lymphocytes with different chemokine receptors at various stages of disease progression. AB - We studied the distribution of human immunodeficiency virus type 1 (HIV-1) DNA in CCR5-positive and -negative peripheral blood lymphocyte populations in HIV-1 infected individuals. While HIV-1 DNA in the CCR5-positive population showed no correlation with CD4 count, the increase of total HIV-1 DNA with lower CD4 count was mainly contributed by the increase of HIV-1 DNA in the CCR5-negative population. This might indicate the change in coreceptor usage from CCR5 to CXCR4 in later stages of disease progression. However, some of the samples with a high viral DNA load in the CCR5-negative population did not have any characteristic of the V3 loop sequence that is compatible with CXCR4 usage or the syncytium inducing (SI) phenotype. We also did not find any known characteristic change predictive of the SI phenotype in V1 and V2 sequences. Our findings showed that there might be a shift in target cell populations during disease progression, and this shift was not necessarily associated with the genetic changes characteristic of CXCR4 usage. PMID- 11413306 TI - In the simian virus 40 in vitro replication system, start site selection by the polymerase alpha-primase complex is not significantly altered by changes in the concentration of ribonucleotides. AB - The simian virus 40 (SV40) in vitro replication system was previously used to demonstrate that the human polymerase (Pol) alpha-primase complex preferentially initiates DNA synthesis at pyrimidine-rich trinucleotide sequences. However, it has been reported that under certain conditions, nucleoside triphosphate (NTP) concentrations play a critical role in determining where eukaryotic primase initiates synthesis. Therefore, we have examined whether increased NTP concentrations alter the template locations at which SV40 replication is initiated. Our studies demonstrate that elevated ribonucleotide concentrations do not significantly alter which template sequences serve as initiation sites. Of considerable interest, the sequences that serve as initiation sites in the SV40 system are similar to those that serve as initiation sites for prokaryotic primases. It is also demonstrated that regardless of the concentration of ribonucleotides present in the reactions, DNA synthesis initiated outside of the core origin. These studies provide additional evidence that the Pol alpha-primase complex can initiate DNA synthesis only after a considerable amount of single stranded DNA is generated. PMID- 11413307 TI - Induction of caspase-dependent apoptosis in cultured cells by the avian coronavirus infectious bronchitis virus. AB - Avian coronavirus infectious bronchitis virus (IBV) is the causative agent of chicken infectious bronchitis, an acute, highly contagious viral respiratory disease. Replication of IBV in Vero cells causes extensive cytopathic effects (CPE), leading to destruction of the entire monolayer and the death of infected cells. In this study, we investigated the cell death processes during acute IBV infection and the underlying mechanisms. The results show that both necrosis and apoptosis may contribute to the death of infected cells in lytic IBV infection. Caspase-dependent apoptosis, as characterized by chromosomal condensation, DNA fragmentation, caspase-3 activation, and poly(ADP-ribose) polymerase degradation, was detected in IBV-infected Vero cells. Addition of the general caspase inhibitor z-VAD-FMK to the culture media showed inhibition of the hallmarks of apoptosis and increase of the release of virus to the culture media at 16 h postinfection. However, neither the necrotic process nor the productive replication of IBV in Vero cells was severely affected by the inhibition of apoptosis. Screening of 11 IBV-encoded proteins suggested that a 58-kDa mature cleavage product could induce apoptotic changes in cells transiently expressing the protein. This study adds one more example to the growing list of animal viruses that induce apoptosis during their replication cycles. PMID- 11413308 TI - Changes in human immunodeficiency virus type 1 populations after treatment interruption in patients failing antiretroviral therapy. AB - Mutations in human immunodeficiency virus type 1 (HIV-1) reverse transcriptase and protease that confer resistance to antiretroviral agents are usually accompanied by a reduction in the viral replicative capacity under drug-free conditions. Consequently, when antiretroviral treatment is interrupted in HIV-1 infected patients harboring drug-resistant virus, resistant quasi-species appear to be most often replaced within several weeks by wild-type virus. Using a real time PCR-based technique for the selective quantification of resistant viral sequences in plasma, we have studied the kinetics of the switch from mutant to wild-type virus and evaluated the extent to which minority populations of resistant viruses not detected by genotyping persist in these individuals. Among 12 patients with viruses expressing the V82A or L90M resistance mutation who had undergone a 3-month interruption of therapy and for whom conventional genotyping had revealed an apparent total reconversion to wild-type virus, minority populations expressing these mutations, representing 0.1 to 21% of total virus, were still detectable in 9 cases. Kinetic studies demonstrated that viruses expressing resistance mutations could be detected for >5 months after the discontinuation of treatment in some patients. Most of the minority resistant genomes detected more than 3 months after the interruption of therapy carried only part of the mutations present in the resistant viruses prior to treatment interruption and appeared to result from the emergence of existing strains selected at earlier stages in the development of drug resistance. Thus, following the interruption of treatment, viral populations containing resistance mutations can persist for several months after the time when conventional genotyping techniques detect only wild-type virus. These populations include viral strains with only some of the resistance mutations initially present, strains that presumably express better fitness under drug-free conditions. PMID- 11413309 TI - The hemagglutinin of canine distemper virus determines tropism and cytopathogenicity. AB - Canine distemper virus (CDV) and measles virus (MV) cause severe illnesses in their respective hosts. The viruses display a characteristic cytopathic effect by forming syncytia in susceptible cells. For CDV, the proficiency of syncytium formation varies among different strains and correlates with the degree of viral attenuation. In this study, we examined the determinants for the differential fusogenicity of the wild-type CDV isolate 5804Han89 (CDV(5804)), the small- and large-plaque-forming variants of the CDV vaccine strain Onderstepoort (CDV(OS) and CDV(OL), respectively), and the MV vaccine strain Edmonston B (MV(Edm)). The cotransfection of different combinations of fusion (F) and hemagglutinin (H) genes in Vero cells indicated that the H protein is the main determinant of fusion efficiency. To verify the significance of this observation in the viral context, a reverse genetic system to generate recombinant CDVs was established. This system is based on a plasmid containing the full-length antigenomic sequence of CDV(OS). The coding regions of the H proteins of all CDV strains and MV(Edm) were introduced into the CDV and MV genetic backgrounds, and recombinant viruses rCDV-H(5804), rCDV-H(OL), rCDV-H(Edm), rMV-H(5804), rMV-H(OL), and rMV-H(OS) were recovered. Thus, the H proteins of the two morbilliviruses are interchangeable and fully functional in a heterologous complex. This is in contrast with the glycoproteins of other members of the family Paramyxoviridae, which do not function efficiently with heterologous partners. The fusogenicity, growth characteristics, and tropism of the recombinant viruses were examined and compared with those of the parental strains. All these characteristics were found to be predominantly mediated by the H protein regardless of the viral backbone used. PMID- 11413310 TI - Multiple cis regulatory elements control RANTES promoter activity in alveolar epithelial cells infected with respiratory syncytial virus. AB - Respiratory syncytial virus (RSV) produces intense pulmonary inflammation, in part through its ability to induce chemokine synthesis in infected airway epithelial cells. RANTES (regulated upon activation, normally T-cell expressed and presumably secreted) is a CC chemokine which recruits and activates monocytes, lymphocytes, and eosinophils, all cell types present in the lung inflammatory infiltrate induced by RSV infection. In this study, we analyzed the mechanism of RSV-induced RANTES promoter activation in human type II alveolar epithelial cells (A549 cells). Promoter deletion and mutagenesis experiments indicate that RSV requires the presence of five different cis regulatory elements, located in the promoter fragment spanning from -220 to +55 nucleotides, corresponding to NF-kappaB, C/EBP, Jun/CREB/ATF, and interferon regulatory factor (IRF) binding sites. Although site mutations of the NF-kappaB, C/EBP, and CREB/AP 1 like sites reduce RSV-induced RANTES gene transcription to 50% or less, only mutations affecting IRF binding completely abolish RANTES inducibility. Supershift and microaffinity isolation assays were used to identify the different transcription factor family members whose DNA binding activity was RSV inducible. Expression of dominant negative mutants of these transcription factors further established their central role in virus-induced RANTES promoter activation. Our finding that the presence of multiple cis regulatory elements is required for full activation of the RANTES promoter in RSV-infected alveolar epithelial cells supports the enhanceosome model for RANTES gene transcription, which is absolutely dependent on binding of IRF transcription factors. The identification of regulatory mechanisms of RANTES gene expression is fundamental for rational design of inhibitors of RSV-induced lung inflammation. PMID- 11413311 TI - Role of the 3'-untranslated regions of alfalfa mosaic virus RNAs in the formation of a transiently expressed replicase in plants and in the assembly of virions. AB - Alfalfa mosaic virus (AMV) RNAs 1 and 2 encode the replicase proteins P1 and P2, respectively, whereas RNA 3 encodes the movement protein and the coat protein (CP). When RNAs 1 and 2 were transiently expressed from a T-DNA vector (R12 construct) by agroinfiltration of Nicotiana benthamiana, the infiltrated leaves accumulated minus-strand RNAs 1 and 2 and relatively small amounts of plus-strand RNAs. In addition, RNA-dependent RNA polymerase (RdRp) activity could be detected in extracts of the infiltrated leaves. After transient expression of RNAs 1 and 2 with the 3'-untranslated regions (UTRs) of both RNAs deleted (R1Delta/2Delta construct), no replication of RNAs 1 and 2 was observed, while the infiltrated leaves supported replication of RNA 3 after inoculation of the leaves with RNA 3 or expression of RNA 3 from a T-DNA vector (R3 construct). No RdRp activity could be isolated from leaves infiltrated with the R1Delta/2Delta construct, although P1 and P2 sedimented in a region of a glycerol gradient where active RdRp was found in plants infiltrated with R12. RdRp activity could be isolated from leaves infiltrated with constructs R1Delta/2 (3'-UTR of RNA 1 deleted), R1/2Delta (3' UTR of RNA 2 deleted), or R1Delta/2Delta plus R3. This demonstrates that the 3' UTR of AMV RNAs is required for the formation of a complex with in vitro enzyme activity. RNAs 1 and 2 with the 3'-UTRs deleted were encapsidated into virions by CP expressed from RNA 3. This shows that the high-affinity binding site for CP at the 3'-termini of AMV RNAs is not required for assembly of virus particles. PMID- 11413312 TI - Adenovirus type 7 induces interleukin-8 production via activation of extracellular regulated kinase 1/2. AB - Infection with adenovirus serotype 7 (Ad7) frequently causes lower respiratory pneumonia and is associated with severe lung inflammation and neutrophil infiltration. Earlier studies indicated release of proinflammatory cytokines, specifically interleukin-8 (IL-8), by pulmonary epithelial cells following infection by Ad7. However, the mechanism of IL-8 induction by Ad7 is unclear. We have explored the role of the Ras/Raf/MEK/Erk pathway in the Ad7-associated induction of IL-8 using a model system of A549 epithelial cells. We found that Ad7 infection induced a rapid activation of epithelial cell-derived Erk. The MEK specific inhibitors PD98059 and U0126 blocked Erk activation and release of IL-8 following infection with Ad7. Treatment with PD98059 is cytostatic and not cytotoxic, as treated cells regain the ability to phosphorylate Erk and secrete IL-8 after removal of the drug. The expression of a mutated form of Ras in A549 epithelial cells blocked the induction of IL-8 promoter activity, and MEK inhibitor blocked induction of IL-8 mRNA. These results suggest that the Ras/Raf/MEK/Erk pathway is necessary for the Ad7 induction of IL-8 and that induction occurs at the level of transcription. Further, the kinetics of Erk activation and IL-8 induction suggest that an early viral event, such as receptor binding, may be responsible for the observed inflammatory response. PMID- 11413313 TI - Repression of vaccinia virus Holliday junction resolvase inhibits processing of viral DNA into unit-length genomes. AB - The vaccinia virus A22R gene encodes a protein that is homologous to the bacterial enzyme RuvC and specifically cleaves and resolves four-way DNA Holliday junctions into linear duplex products. To investigate the role of the vaccinia virus Holliday junction resolvase during an infection, we constructed two recombinant viruses: vA22-HA, which has a short C-terminal epitope tag appended to the A22R open reading frame, and vA22i, in which the original A22R gene is deleted and replaced by an inducible copy. Polyacrylamide gel electrophoresis and Western blot analysis of extracts and purified virions from cells infected with vA22-HA revealed that the resolvase was expressed after the onset of DNA replication and incorporated into virion cores. vA22i exhibited a conditional replication defect. In the absence of an inducer, (i) viral protein synthesis was unaffected, (ii) late-stage viral DNA replication was reduced, (iii) most of the newly synthesized viral DNA remained in a branched or concatemeric form that caused it to be trapped at the application site during pulsed-field gel electrophoresis, (iv) cleavage of concatemer junctions was inhibited, and (v) virion morphogenesis was arrested at an immature stage. These data indicated multiple roles for the vaccinia virus Holliday junction resolvase in the replication and processing of viral DNA into unit-length genomes. PMID- 11413314 TI - A predicted secondary structural domain within the internal ribosome entry site of echovirus 12 mediates a cell-type-specific block to viral replication. AB - The enterovirus 5' nontranslated region (NTR) contains an internal ribosome entry site (IRES), which facilitates translation initiation of the viral open reading frame in a 5' (m(7)GpppN) cap-independent manner, and cis-acting signals for positive-strand RNA replication. For several enteroviruses, the 5' NTR has been shown to determine the virulence phenotype. We have constructed a chimera consisting of the putative IRES element from the Travis strain of echovirus 12 (ECV12), a wild-type, relatively nonvirulent human enterovirus, exchanged with the homologous region of a full-length infectious clone of coxsackievirus B3 (CBV3). The resulting chimera, known as ECV12(5'NTR)CBV3, replicates similarly to CBV3 in human and simian cell lines yet, unlike CBV3, is completely restricted for growth on two primary murine cell lines at 37 degrees C. By utilizing a reverse-genetics approach, the growth restriction phenotype was localized to the predicted stem-loop II within the IRES of ECV12. In addition, a revertant of ECV12(5'NTR)CBV3 was isolated which possessed three transition mutations and had restored capability for replication in the utilized murine cell lines. Assays for cardiovirulence indicated that the ECV12 IRES is responsible for a noncardiovirulent phenotype in a murine model for acute myocarditis. The results indicate that the 5' NTRs of ECV12 and CBV3 exhibit variable intracellular requirements for function and serve as secondary determinants of tissue or species tropism. PMID- 11413315 TI - CpG oligodeoxynucleotides with hepatitis B surface antigen (HBsAg) for vaccination in HBsAg-transgenic mice. AB - DNA motifs containing unmethylated CpG dinucleotides within the context of certain flanking sequences enhance both innate and antigen-specific immune responses, due in part to the enhanced production of Th1-type cytokines. Here we explored the ability of CpG-containing oligodeoxynucleotides combined with recombinant hepatitis B surface antigen (HBsAg) to induce Th1 responses in mice that are transgenic for this antigen and that represent a model for asymptomatic hepatitis B virus chronic carriers. This was compared to hepatitis B virus specific DNA-mediated immunization, which we have previously shown to induce the clearance of the transgene expression product and the down-regulation of hepatitis B virus mRNA in this transgenic mouse lineage. In control nontransgenic C57BL/6 mice, three immunizations with HBsAg and CpG triggered the production of anti-HBs antibodies and of HBs-specific T cells that secrete gamma interferon but do not display any HBsAg-specific cytotoxic activity. In the HBsAg-transgenic mice, immunization with HBsAg and CpG oligodeoxynucleotides, but not with CpG alone, induced the clearance of HBsAg circulating in the sera, with a concomitant appearance of specific antibodies, and was able to regulate the hepatitis B virus mRNA constitutively expressed in the liver. Finally, adoptive transfer experiments with CD8(+) T cells primed in C57BL/6 mice with HBsAg and CpG oligodeoxynucleotide-based immunization show that these cells were able to partially control transgene expression in the liver and to clear the HBsAg from the sera of recipient transgenic mice without an antibody requirement. CpG oligodeoxynucleotides motifs combined with HBsAg could therefore represent a potential therapeutic approach with which to treat chronically infected patients. PMID- 11413316 TI - Mutations that increase in situ priming also decrease circularization for duck hepatitis B virus. AB - The process of hepadnavirus reverse transcription involves two template switches during the synthesis of plus-strand DNA. The first involves translocation of the plus-strand primer from its site of generation, the 3' end of minus-strand DNA, to the complementary sequence DR2, located near the 5' end of the minus-strand DNA. Plus strands initiated from DR2 are extended to the 5' end of the minus strand DNA. At this point, the 3' end of the minus strand becomes the template via the second template switch, a process called circularization. Elongation of circularized plus-strand DNA generates relaxed circular DNA. Although most virions contain relaxed circular DNA, some contain duplex linear DNA. Duplex linear genomes are synthesized when the plus-strand primer is used at the site of its generation, the 3' end of the minus-strand template. This type of synthesis is called in situ priming. Although in situ priming is normally low, in some duck hepatitis B virus mutants this type of priming is elevated. For example, mutations within the 3' end of the minus-strand DNA can lead to increased levels of in situ priming. We report here that these same mutations result in a second defect, a less efficient template switch that circularizes the genome. Although it is not clear how these mutations affect both steps in DNA replication, our findings suggest a commonality in the mechanism of initiation of plus-strand synthesis and the template switch that circularizes the genome. PMID- 11413317 TI - Transactivation of murine cyclin A by polyomavirus large and small T antigens. AB - Polyomavirus large and small T antigens cooperate in the induction of S phase in serum-deprived Swiss 3T3 cells. While the large T antigen is able to induce S phase-specific enzymes, we have recently shown that both T antigens contribute to the production of the cyclins E and A and that the small T antigen is essential for the induction of cyclin A-dependent cdk2 activity (S. Schuchner and E. Wintersberger, J. Virol. 73:9266-9273, 1999). Here we present our attempts to elucidate the mechanisms by which the large and the small T antigens transactivate the murine cyclin A gene. Using Swiss 3T3 cells carrying the T antigens and various mutants thereof under the hormone-inducible mouse mammary tumor virus promoter, as well as transient-cotransfection experiments with the T antigens and cyclin A promoter-luciferase reporter constructs, we found the following. The large T antigen activates the cyclin A promoter via two transcription factor binding sites, a cyclic AMP responsive element (CRE), and the major negative regulatory site called CDE-CHR. While an intact binding site for pocket proteins is required for the function of this T antigen at the CDE CHR, its activity at the CRE is largely independent thereof. In contrast, an intact J domain and an intact zinc finger are required at both sites. The small T antigen also appears to have an influence on the cyclin A promoter through the CRE as well as the CDE-CHR. For this an interaction with protein phosphatase 2A is essential; mutation of the J domain does not totally eliminate but greatly reduces the transactivating ability. PMID- 11413318 TI - Decay kinetics of human immunodeficiency virus-specific CD8+ T cells in peripheral blood after initiation of highly active antiretroviral therapy. AB - We measured the longitudinal responses to 95 HLA class I-restricted human immunodeficiency virus (HIV) epitopes and an immunodominant HLA A2-restricted cytomegalovirus (CMV) epitope in eight treatment-naive HIV-infected individuals, using intracellular cytokine staining. Patients were treated with highly active antiretroviral therapy (HAART) for a median of 78 weeks (range, 34 to 121 weeks). Seven of eight patients maintained an undetectable viral load for the duration of therapy. A rapid decline in HIV-specific CD8(+) T-cell response was observed at initiation of therapy. After an undetectable viral load was achieved, a slower decrease in HIV-specific CD8(+) T-cell response was observed that was well described by first-order kinetics. The median half-life for the rate of decay was 38.8 (20.3 to 68.0) weeks when data were expressed as percentage of peripheral CD8(+) T cells. In most cases, data were similar when expressed as the number of responding CD8(+) T cells per microliter of blood. In subjects who responded to more than one HIV epitope, rates of decline in response to the different epitopes were similar and varied by a factor of 2.2 or less. Discontinuation of treatment resulted in a rapid increase in HIV-specific CD8(+) T cells. Responses to CMV increased 1.6- and 2.8-fold within 16 weeks of initiation of HAART in two of three patients with a measurable CMV response. These data suggest that HAART quickly starts to restore CD8(+) T-cell responses to other chronic viral infections and leads to a slow decrease in HIV-specific CD8(+) T-cell response in HIV-infected patients. The slow decrease in the rate of CD8(+) T-cell response and rapid increase in response to recurrent viral replication suggest that the decrease in CD8(+) T-cell response observed represents a normal memory response to withdrawal of antigen. PMID- 11413319 TI - The reovirus S4 gene 3' nontranslated region contains a translational operator sequence. AB - Reovirus mRNAs are efficiently translated within host cells despite the absence of 3' polyadenylated tails. The 3' nontranslated regions (3'NTRs) of reovirus mRNAs contain sequences that exhibit a high degree of gene-segment-specific conservation. To determine whether the 3'NTRs of reovirus mRNAs serve to facilitate efficient translation of viral transcripts, we used T7 RNA polymerase to express constructs engineered with full-length S4 gene cDNA or truncation mutants lacking sequences in the 3'NTR. Full-length and truncated s4 mRNAs were translated using rabbit reticulocyte lysates, and translation product sigma3 was quantitated by phosphorimager analysis. In comparison to full-length s4 mRNA, translation of the s4 mRNA lacking the 3'NTR resulted in a 20 to 50% decrease in sigma3 produced. Addition to translation reactions of an RNA oligonucleotide corresponding to the S4 3'NTR significantly enhanced translation of full-length s4 mRNA but had no effect on s4 mRNA lacking 3'NTR sequences. Translation of s4 mRNAs with smaller deletions within the 3'NTR identified a discrete region capable of translational enhancement and a second region capable of translational repression. Differences in translational efficiency of full-length and deletion mutant mRNAs were independent of RNA stability. Protein complexes in reticulocyte lysates that specifically interact with the S4 3'NTR were identified by RNA mobility shift assays. RNA oligonucleotides lacking either enhancer or repressor sequences did not efficiently compete the binding of these complexes to full length 3'NTR. These results indicate that the reovirus S4 gene 3'NTR contains a translational operator sequence that serves to regulate translational efficiency of the s4 mRNA. Moreover, these findings suggest that cellular proteins interact with reovirus 3'NTR sequences to regulate translation of the nonpolyadenylated reovirus mRNAs. PMID- 11413320 TI - Insertion of capsid proteins from nonenveloped viruses into the retroviral budding pathway. AB - Retroviral Gag proteins direct the assembly and release of virus particles from the plasma membrane. The budding machinery consists of three small domains, the M (membrane-binding), I (interaction), and L (late or "pinching-off") domains. In addition, Gag proteins contain sequences that control particle size. For Rous sarcoma virus (RSV), the size determinant maps to the capsid (CA)-spacer peptide (SP) sequence, but it functions only when I domains are present to enable particles of normal density to be produced. Small deletions throughout the CA-SP sequence result in the release of particles that are very large and heterogeneous, even when I domains are present. In this report, we show that particles of relatively uniform size and normal density are released by budding when the size determinant and I domains in RSV Gag are replaced with capsid proteins from two unrelated, nonenveloped viruses: simian virus 40 and satellite tobacco mosaic virus. These results indicate that capsid proteins of nonenveloped viruses can interact among themselves within the context of Gag and be inserted into the retroviral budding pathway merely by attaching the M and L domains to their amino termini. Thus, the differences in the assembly pathways of enveloped and nonenveloped viruses may be far simpler than previously thought. PMID- 11413321 TI - Replication of phenotypically mixed human immunodeficiency virus type 1 virions containing catalytically active and catalytically inactive reverse transcriptase. AB - The amount of excess polymerase and RNase H activity in human immunodeficiency virus type 1 virions was measured by using vectors that undergo a single round of replication. Vectors containing wild-type reverse transcriptase (RT), vectors encoding the D110E mutation to inactivate polymerase, and vectors encoding mutations D443A and E478Q to inactivate RNase H were constructed. 293 cells were cotransfected with different proportions of plasmids encoding these vectors to generate phenotypically mixed virions. The resulting viruses were used to infect human osteosarcoma cells, and the relative infectivity of the viruses was determined by measuring transduction of the murine cell surface marker CD24, which is encoded by the vectors. The results indicated that there is an excess of both polymerase and RNase H activities in virions. Viral replication was reduced to 42% of wild-type levels in virions with where half of the RT molecules were predicted to be catalytically active but dropped to 3% of wild-type levels when 25% of the RT molecules were active. However, reducing RNase H activity had a lesser effect on viral replication. As expected, based on previous work with murine leukemia virus, there was relatively inefficient virus replication when the RNase H and polymerase activities were encoded on separate vectors (D110E plus E478Q and D110E plus D443A). To determine how virus replication failed when polymerase and RNase H activities were reduced, reverse transcription intermediates were measured in vector-infected cells by using quantitative real time PCR. The results indicated that using the D11OE mutation to reduce the amount of active polymerase reduced the number of reverse transcripts that were initiated and also reduced the amounts of products from the late stages of reverse transcription. If the E478Q mutation was used to reduce RNase H activity, the number of reverse transcripts that were initiated was reduced; there was also a strong effect on minus-strand transfer. PMID- 11413322 TI - Cell-type-specific regulation of the two foamy virus promoters. AB - The foamy virus (FV) genome contains two promoters, the canonical long terminal repeat (LTR) promoter, containing three consensus AP-1 binding sites, and an internal promoter (IP) within the env gene. We investigated the regulation of the two promoters in lytic and persistent infections and found that in the presence of a constitutive source of the viral transactivator protein Tas, transactivation of the LTR promoter and that of the IP differ. In lytic infections, both the LTR promoter and the IP are efficiently transactivated by Tas, while in persistent infections, the IP is efficiently transactivated by Tas, but the LTR promoter is not. Analysis of proteins expressed from the LTR promoter and the IP during infection indicated that IP transcription is more robust than that of the LTR promoter in persistently infected cells, while the opposite is true for lytically infected cells. Coculture experiments also showed that LTR promoter transcription is greatest in cells which support lytic replication. Replacement of much of the LTR promoter with the IP leads to increased viral replication in persistent but not lytic infections. We also found that the induction of persistently infected cells with phorbol 12-myristate 13-acetate (PMA) greatly enhanced viral replication and transcription from the SFVcpz(hu) (new name for human FV) LTR promoter. However, mutation of three consensus AP-1 binding sites in the FV LTR promoter did not affect viral replication in lytically or persistently infected cells, nor did the same mutations affect LTR promoter transactivation by Tas in PMA-treated cells. Our data indicate that differential regulation of transcription is important in the outcome of FV infection but is unlikely to depend on AP-1. PMID- 11413324 TI - Genetic diversity in RNA virus quasispecies is controlled by host-virus interactions. AB - Many RNA viruses have genetically diverse populations known as quasispecies. Important biological characteristics may be related to the levels of diversity in the quasispecies (quasispecies cloud size), including adaptability and host range. Previous work using Tobacco mosaic virus and Cucumber mosaic virus indicated that evolutionarily related viruses have very different levels of diversity in a common host. The quasispecies cloud size for these viruses remained constant throughout serial passages. Inoculation of these viruses on a number of hosts demonstrated that quasispecies cloud size is not constant for these viruses but appears to be dependent on the host. The quasispecies cloud size remained constant as long as the viruses were maintained on a given host. Shifting the virus between hosts resulted in a change in cloud size to levels associated with the new host. Quasispecies cloud size for these viruses is related to host-virus interactions, and understanding these interactions may facilitate the prediction and prevention of emerging viral diseases. PMID- 11413323 TI - Immunoglobulin G3 from polyclonal human immunodeficiency virus (HIV) immune globulin is more potent than other subclasses in neutralizing HIV type 1. AB - Passive antibody prophylaxis against human immunodeficiency virus type 1 (HIV-1) has been accomplished in primates, suggesting that this strategy may prove useful in humans. While antibody specificity is crucial for neutralization, other antibody characteristics, such as subclass, have not been explored. Our objective was to compare the efficiencies of immunoglobulin G (IgG) subclasses from polyclonal human HIV immune globulin (HIVIG) in the neutralization of HIV-1 strains differing in coreceptor tropism. IgG1, IgG2, and IgG3 were enriched from HIVIG by using protein A-Sepharose. All three subclasses bound major HIV-1 proteins, as shown by Western blot assay and enzyme-linked immunosorbent assay. In HIV-1 fusion assays using X4, R5, or X4R5 envelope-expressing effector cells, IgG3 more efficiently blocked fusion. In neutralization assays with cell-free viruses using X4 (LAI, IIIB), R5 (BaL), and X4R5 (DH123), a similar hierarchy of neutralization was found: IgG3 > IgG1 > IgG2. IgG3 has a longer, more flexible hinge region than the other subclasses. To test whether this is important, IgG1 and IgG3 were digested with pepsin to generate F(ab')(2) fragments or with papain to generate Fab fragments. IgG3 F(ab')(2) fragments were still more efficient in neutralization than F(ab')(2) of IgG1. However, Fab fragments of IgG3 and IgG1 demonstrated equivalent neutralization capacities and the IgG3 advantage was lost. These results suggest that the IgG3 hinge region confers enhanced HIV neutralizing ability. Enrichment and stabilization of IgG3 may therefore lead to improved HIVIG preparations. The results of this study have implications for the improvement of passive immunization with polyclonal or monoclonal antibodies and suggest that HIV-1 vaccines which induce high-titer IgG3 responses could be advantageous. PMID- 11413326 TI - Experimental preemptive immunotherapy of murine cytomegalovirus disease with CD8 T-cell lines specific for ppM83 and pM84, the two homologs of human cytomegalovirus tegument protein ppUL83 (pp65). AB - CD8 T cells are the principal antiviral effectors controlling cytomegalovirus (CMV) infection. For human CMV, the virion tegument protein ppUL83 (pp65) has been identified as a source of immunodominant peptides and is regarded as a candidate for cytoimmunotherapy and vaccination. Two sequence homologs of ppUL83 are known for murine CMV, namely the virion protein ppM83 (pp105) expressed late in the viral replication cycle and the nonstructural protein pM84 (p65) expressed in the early phase. Here we show that ppM83, unlike ppUL83, is not delivered into the antigen presentation pathway after virus penetration before or in absence of viral gene expression, while other virion proteins of murine CMV are processed along this route. In cytokine secretion-based assays, ppM83 and pM84 appeared to barely contribute to the acute immune response and to immunological memory. Specifically, the frequencies of M83 and M84 peptide-specific CD8 T cells were low and undetectable, respectively. Nonetheless, in a murine model of cytoimmunotherapy of lethal CMV disease, M83 and M84 peptide-specific cytolytic T cell lines proved to be highly efficient in resolving productive infection in multiple organs of cell transfer recipients. These findings demonstrate that proteins which fail to prime a quantitatively dominant immune response can nevertheless represent relevant antigens in the effector phase. We conclude that quantitative and qualitative immunodominance are not necessarily correlated. As a consequence of these findings, there is no longer a rationale for considering T cell abundance as the key criterion for choosing specificities to be included in immunotherapy and immunoprophylaxis of CMV disease and of viral infections in general. PMID- 11413325 TI - Mononuclear phagocyte differentiation, activation, and viral infection regulate matrix metalloproteinase expression: implications for human immunodeficiency virus type 1-associated dementia. AB - The pathogenesis of human immunodeficiency virus type 1 (HIV-1)-associated dementia (HAD) is mediated mainly by mononuclear phagocyte (MP) secretory products and their interactions with neural cells. Viral infection and MP immune activation may affect leukocyte entry into the brain. One factor that influences central nervous system (CNS) monocyte migration is matrix metalloproteinases (MMPs). In the CNS, MMPs are synthesized by resident glial cells and affect the integrity of the neuropil extracellular matrix (ECM). To ascertain how MMPs influence HAD pathogenesis, we studied their secretion following MP differentiation, viral infection, and cellular activation. HIV-1-infected and/or immune-activated monocyte-derived macrophages (MDM) and human fetal microglia were examined for production of MMP-1, -2, -3, and -9. MMP expression increased significantly with MP differentiation. Microglia secreted high levels of MMPs de novo that were further elevated following CD40 ligand-mediated cell activation. Surprisingly, HIV-1 infection of MDM led to the down-regulation of MMP-9. In encephalitic brain tissue, MMPs were expressed within perivascular and parenchymal MP, multinucleated giant cells, and microglial nodules. These data suggest that MMP production in MP is dependent on cell type, differentiation, activation, and/or viral infection. Regulation of MMP expression by these factors may contribute to neuropil ECM degradation and leukocyte migration during HAD. PMID- 11413327 TI - Nef from human immunodeficiency virus type 1(F12) inhibits viral production and infectivity. AB - Human immunodeficiency virus type 1(F12) (HIV-1(F12)) interferes with the replication of other strains of HIV. Its accessory protein, Nef, is sufficient for this phenotype, where the production and infectivity of HIV are impaired significantly. The analysis of three rare mutations in this Nef protein revealed that these effects could be separated genetically. Moreover, the defect in virus production correlated with the lack of processing of the p55(Gag) precursor in the presence of Nef from HIV-1(F12). Importantly, the introduction of one of these mutations (E177G) into Nef from HIV-1(NL4-3) also created a dominant negative Nef protein. Effects of Nef from HIV-1(F12) on virus production and Gag processing correlated with its altered subcellular distribution. Moreover, the association with two new cellular proteins with molecular masses of 74 and 75 kDa, which do not interact with other Nef proteins, correlated with the decreased virion infectivity. The identification of a dominant-negative protein for the production and infectivity of HIV suggests that Nef plays an active role at this stage of the viral replicative cycle. PMID- 11413328 TI - Recessive resistance in Pisum sativum and potyvirus pathotype resolved in a gene for-cistron correspondence between host and virus. AB - Pea seed-borne mosaic potyvirus (PSbMV) isolates are divided into pathotypes P-1, P-2, and P-4 according to their infection profile on a panel of Pisum sativum lines. P. sativum PI 269818 is resistant to P-1 and P-2 isolates and is susceptible to P-4 isolates. Resistance to P-1 is inherited as a single recessive gene, denoted sbm-1, and the pathogenicity determinant has previously been mapped to the virus-coded protein VPg. In the cultivar Bonneville, a second recessive gene, sbm-2, confers specific resistance to P-2. By exchanging cistrons between a P-2 and a P-4 isolate, the P3-6k1 cistron was identified as the PSbMV host specific pathogenicity determinant on Bonneville. Exchange of P3-6k1 did not affect infection on PI 269818, and infection of Bonneville was not altered by substitution of the VPg cistron, indicating that P3-6k1 and VPg are independent determinants of pathotype-specific infectivity. On PI 269818 the pathogenicity determinant of both P-1 and P-2 mapped to the N terminus of VPg. This suggests that VPg from the P-1 and P-2 isolates are functionally similar on this host and that resistance to P-1 and P-2 in PI 269818 may operate by the same mechanism. Identification of VPg-sbm-1 and P3-6k1-sbm-2 as independent pairs of genetic interactors between PSbMV and P. sativum provides a simple explanation of the three known pathotypes of PSbMV. Furthermore, analysis of beta-glucuronidase tagged P-2 virus indicated that sbm-2 resistance affected an early step in infection, implying that the P3-6k1 region plays a critical role in potyvirus replication or cell-to-cell movement. PMID- 11413329 TI - Adeno-associated virus type 6 (AAV6) vectors mediate efficient transduction of airway epithelial cells in mouse lungs compared to that of AAV2 vectors. AB - Although vectors derived from adeno-associated virus type 2 (AAV2) promote gene transfer and expression in many somatic tissues, studies with animal models and cultured cells show that the apical surface of airway epithelia is resistant to transduction by AAV2 vectors. Approaches to increase transduction rates include increasing the amount of vector and perturbing the integrity of the epithelia. In this study, we explored the use of vectors based on AAV6 to increase transduction rates in airways. AAV vectors were made using combinations of rep, cap, and packaged genomes from AAV2 or AAV6. The packaged genomes encoded human placental alkaline phosphatase and contained terminal repeat sequences from AAV2 or AAV6. We found that transduction efficiency was primarily dependent on the source of Cap protein, defined here as the vector pseudotype. The AAV6 and AAV2 pseudotype vectors exhibited different tropisms in tissue-cultured cells, and cell transduction by AAV6 vectors was not inhibited by heparin, nor did they compete for entry in a transduction assay, indicating that AAV6 and AAV2 capsid bind different receptors. In vivo analysis of vectors showed that AAV2 pseudotype vectors gave high transduction rates in alveolar cells but much lower rates in the airway epithelium. In contrast, the AAV6 pseudotype vectors exhibited much more efficient transduction of epithelial cells in large and small airways, showing up to 80% transduction in some airways. These results, combined with our previous results showing lower immunogenicity of AAV6 than of AAV2 vectors, indicate that AAV6 vectors may provide significant advantages over AAV2 for gene therapy of lung diseases like cystic fibrosis. PMID- 11413330 TI - A monoclonal antibody specific for reovirus outer-capsid protein sigma3 inhibits sigma1-mediated hemagglutination by steric hindrance. AB - Reovirus virions are nonenveloped icosahedral particles consisting of two concentric protein shells, termed outer capsid and core. Outer-capsid protein sigma1 is the viral attachment protein and binds carbohydrate molecules on the surface of host cells. Monoclonal antibody (MAb) 4F2, which is specific for outer capsid protein sigma3, blocks the binding of sigma1 protein to sialic acid and inhibits reovirus-induced hemagglutination (HA). To determine whether MAb 4F2 inhibits HA by altering sigma1-sigma3 interactions or by steric hindrance, we analyzed the effect of 4F2 immunoglobulin G (IgG) and Fab fragments (Fabs) on HA induced by reovirus strain type 3 Dearing (T3D). The concentration of 4F2 IgG sufficient to inhibit T3D-induced HA was 12.5 microg per ml, whereas that of Fabs was >200 microg per ml. Dynamic light scattering analysis showed that at the concentration of IgG sufficient to inhibit HA, virion-antibody complexes were monodispersed and not aggregated. The affinity of 4F2 Fabs for T3D virions was only threefold less than that of intact IgG, which suggests that differences in HA inhibition titer exhibited by 4F2 IgG and Fabs are not attributable to differences in the affinity of these molecules for T3D virions. We used cryoelectron microscopy and three-dimensional image analysis to visualize T3D virions alone and in complex with either IgG or Fabs of MAb 4F2. IgG and Fabs bind the same site at the distal portion of sigma3, and binding of IgG and Fabs induces identical conformational changes in outer-capsid proteins sigma3 and mu1. These results suggest that MAb 4F2 inhibits reovirus binding to sialic acid by steric hindrance and provide insight into the conformational flexibility of reovirus outer-capsid proteins. PMID- 11413331 TI - Functional analysis of the disulfide-bonded loop/chain reversal region of human immunodeficiency virus type 1 gp41 reveals a critical role in gp120-gp41 association. AB - Human immunodeficiency virus type 1 (HIV-1) entry into cells is mediated by the surface-exposed envelope protein (SU) gp120, which binds to cellular CD4 and chemokine receptors, triggering the membrane fusion activity of the transmembrane (TM) protein gp41. The core of gp41 comprises an N-terminal triple-stranded coiled coil and an antiparallel C-terminal helical segment which is packed against the exterior of the coiled coil and is thought to correspond to a fusion activated conformation. The available gp41 crystal structures lack the conserved disulfide-bonded loop region which, in human T-lymphotropic virus type 1 (HTLV-1) and murine leukemia virus TM proteins, mediates a chain reversal, connecting the antiparallel N- and C-terminal regions. Mutations in the HTLV-1 TM protein gp21 disulfide-bonded loop/chain reversal region adversely affected fusion activity without abolishing SU-TM association (A. L. Maerz, R. J. Center, B. E. Kemp, B. Kobe, and P. Poumbourios, J. Virol. 74:6614-6621, 2000). We now report that in contrast to our findings with HTLV-1, conservative substitutions in the HIV-1 gp41 disulfide-bonded loop/chain reversal region abolished association with gp120. While the mutations affecting gp120-gp41 association also affected cell cell fusion activity, HIV-1 glycoprotein maturation appeared normal. The mutant glycoproteins were processed, expressed at the cell surface, and efficiently immunoprecipitated by conformation-dependent monoclonal antibodies. The gp120 association site includes aromatic and hydrophobic residues on either side of the gp41 disulfide-bonded loop and a basic residue within the loop. The HIV-1 gp41 disulfide-bonded loop/chain reversal region is a critical gp120 contact site; therefore, it is also likely to play a central role in fusion activation by linking CD4 plus chemokine receptor-induced conformational changes in gp120 to gp41 fusogenicity. These gp120 contact residues are present in diverse primate lentiviruses, suggesting conservation of function. PMID- 11413332 TI - Biologic studies of chimeras of highly and moderately virulent molecular clones of simian immunodeficiency virus SIVsmPBj suggest a critical role for envelope in acute AIDS virus pathogenesis. AB - Previous studies identified three molecular clones of the acutely pathogenic SIVsmPBj strain that varied in terms of relative in vivo pathogenicity. One clone, SIVsmPBj6.6, reproducibly induced a rapidly fatal disease in pigtailed macaques. In contrast, a highly related clone (SIVsmPBj6.9) was only minimally pathogenic in macaques. PBj6.6 and PBj6.9 shared a tyrosine substitution at position 17 in the Nef protein that is a major determinant of virulence but differed at one residue in Vpx (C89R), three residues within the envelope (D119G, R871G, G872R), and a single residue in Nef (F252L). SIVsmPBj6.9 was less efficient in inducing proliferation of resting macaque peripheral blood mononuclear cells in vitro than SIVsmPBj6.6 and exhibited a marked reduction in infectivity relative to SIVsmPBj6.6. Chimeric viruses for each of these variable residues were constructed, and their biologic properties were compared to those of the parental strains. Differences in Vpx and Nef did not alter the basic biologic phenotype of the chimeras. However, the D119G substitution in the envelope of SIVsmPBj6.9 was associated with a marked reduction in the infectivity of this virus relative to SIVsmPBj6.6. An associated processing defect in gp160 of SIVsmPBj6.9 and chimeras expressing the D119G substitution suggests that a reduction in virion envelope incorporation is the mechanistic basis for reduced virion infectivity. In vivo studies revealed that substitution of the PBj6.9 amino acid into PBj6.6 (D119) abrogated the pathogenicity of this previously pathogenic virus. Introduction of the PBj6.9 G119, however, did not confer full virulence to the parental PBj6.9 virus, implicating one or all of the other four substitutions in the virulence of SIVsmPBj6.6. PMID- 11413333 TI - Herpes simplex virus type 1 latency-associated transcript gene promotes neuronal survival. AB - A complex interaction has evolved between the host's peripheral nervous system (PNS) and herpes simplex virus type 1 (HSV-1). Sensory neurons are permissive for viral replication, yet the virus can also enter a latent state in these cells. The interplay of viral and neuronal signals that regulate the switch between the viral lytic and latent states is not understood. The latency-associated transcript (LAT) regulates the establishment of the latent state and is required for >65% of the latent infections established by HSV-1 (R. L. Thompson and N. M. Sawtell, J. Virol. 71:5432-5440, 1997). To further investigate how LAT functions, a 1.9-kb deletion that includes the entire LAT promoter and 827 bp of the 5' end of the primary LAT mRNA was introduced into strain 17syn+. The wild-type parent, three independently derived deletion mutants, and two independently derived genomically rescued variants of the mutants were analyzed in a mouse ocular model. The number of latent sites established in trigeminal ganglion (TG) neurons was determined using a single-cell quantitative PCR assay for the viral genome on purified TG neurons. It was found that the LAT null mutants established ~75% fewer latent infections than the number established by the parental strain or rescued variant. The reduced establishment phenotype of LAT null mutants was due at least in part to a dramatic increase in the loss of TG neurons in animals infected with the LAT mutants. Over half of the neurons in the TG were destroyed following infection with the LAT mutants, and this was significantly more than were lost following infection with wild type. This is the first demonstration that the HSV LAT locus prevents the destruction of sensory neurons. The death of these neurons did not appear to be the result of increased apoptosis as measured by a terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling assay. Animals latently infected with the LAT null mutants reactivated less frequently in vivo and this was consistent with the reduction in the number of neurons in which latency was established. Thus, one function of the LAT gene is to protect sensory neurons and enhance the establishment of latency in the PNS. PMID- 11413334 TI - Viral replicase gene products suffice for coronavirus discontinuous transcription. AB - We have used vaccinia virus as a vector to clone a 22.5-kbp cDNA that represents the 5' and 3' ends of the human coronavirus 229E (HCoV 229E) genome, the HCoV 229E replicase gene, and a single reporter gene (coding for green fluorescent protein [GFP]) located downstream of a regulatory element for coronavirus mRNA transcription. When RNA transcribed from this cDNA was transfected into BHK-21 cells, a small percentage of cells displayed strong fluorescence. A region of the mRNA encoding GFP was amplified by PCR and shown to have the unique mRNA leader body junction indicative of coronavirus-mediated transcription. These data show that the coronavirus replicase gene products suffice for discontinuous subgenomic mRNA transcription. PMID- 11413335 TI - Structural flexibility and functional valence of CD4-IgG2 (PRO 542): potential for cross-linking human immunodeficiency virus type 1 envelope spikes. AB - CD4-immunoglobulin G2 (CD4-IgG2) incorporates four copies of the D1D2 domains of CD4 into an antibody-like molecule that potently neutralizes primary human immunodeficiency virus type 1. Here electron microscopy was used to explore the structure and functional valence of CD4-IgG2 in complex with gp120. CD4-gamma2, a divalent CD4-immunoglobulin fusion protein, was evaluated in parallel. Whereas CD4-gamma2-gp120 complexes adopted a simple Y-shaped structure, CD4-IgG2-gp120 complexes consisted of four gp120s arrayed about a central CD4-IgG2 molecule, a structure more reminiscent of complement C1q. Molecular modeling corroborated the electron microscopy data and further indicated that CD4-IgG2 but not CD4-gamma2 has significant potential to cross-link gp120-gp41 trimers on the virion surface, suggesting a mechanism for the heightened antiviral activity of CD4-IgG2. PMID- 11413336 TI - Pathogenesis of influenza A (H5N1) virus infection in a primate model. AB - Cynomolgus macaques (Macaca fascicularis) infected with influenza virus A/Hong Kong/156/97 (H5N1) developed acute respiratory distress syndrome and fever associated with a necrotizing interstitial pneumonia. Reverse transcription PCR, virus isolation, and immunohistochemistry showed that the respiratory tract is the major target of the virus. PMID- 11413337 TI - Identification and characterization of a peptide that specifically binds the human, broadly neutralizing anti-human immunodeficiency virus type 1 antibody b12. AB - Human monoclonal antibody (MAb) b12 recognizes a conformational epitope that overlaps the CD-4-binding site of the human immunodeficiency virus type 1 (HIV-1) envelope. MAb b12 neutralizes a broad range of HIV-1 primary isolates and protects against primary virus challenge in animal models. We report here the discovery and characterization of B2.1, a peptide that binds specifically to MAb b12. B2.1 was selected from a phage-displayed peptide library by using immunoglobulin G1 b12 as the selecting agent. The peptide is a homodimer whose activity depends on an intact disulfide bridge joining its polypeptide chains. Competition studies with gp120 indicate that B2.1 occupies the b12 antigen binding site. The affinity of b12 for B2.1 depends on the form in which the peptide is presented; b12 binds best to the homodimer as a recombinant polypeptide fused to the phage coat. Originally, b12 was isolated from a phage displayed Fab library constructed from the bone marrow of an HIV-1-infected donor. The B2.1 peptide is highly specific for b12 since it selected only phage bearing b12 Fab from this large and diverse antibody library. PMID- 11413338 TI - Induction and characterization of neutralizing antibodies against a human immunodeficiency virus type 1 primary isolate. AB - Chimpanzees infected with the primary isolate DH012 mount potent neutralizing antibodies. This DH012 neutralizing activity is highly strain specific. Immune sera from guinea pigs immunized with recombinant DH012 gp120 could also neutralize this primary isolate. The neutralizing activity in chimpanzee and guinea pig sera against wild-type DH012 appears to be independent of a linear epitope in the V3 region of gp120. Interestingly, the neutralization escape mutant derived from growing DH012 in the presence of the potent neutralizing chimpanzee serum is at least 50-fold more sensitive than wild-type DH012 to neutralization by guinea pig immune sera. The unusually potent neutralizing activity against the DH012 neutralization-resistant virus is due to the presence of anti-V3 antibodies in guinea pig sera. These results suggested that recombinant gp120 could induce neutralizing antibodies against primary isolate DH012. The V3 of wild-type DH012 is poorly immunogenic in infected chimpanzees and is not accessible to neutralizing V3 antibodies. It is likely that this cryptic V3 region became exposed when the virus escaped the neutralizing activity of the chimpanzee serum. PMID- 11413339 TI - Interleukin-12 (IL-12) and IL-18 are important in innate defense against genital herpes simplex virus type 2 infection in mice but are not required for the development of acquired gamma interferon-mediated protective immunity. AB - Using a combination of gene-targeted mice and neutralizing antibodies, we showed that interleukin-12 (IL-12) and IL-18 are important in the innate control of genital herpes simplex virus type 2 infection but were not found to be critical, either singly or in combination, for the development of a protective gamma interferon-mediated immune response. PMID- 11413340 TI - Plasmacytoid dendritic cells are highly susceptible to human immunodeficiency virus type 1 infection and release infectious virus. AB - Plasmacytoid dendritic cells (pcDC) and myeloid dendritic cells (myDC) are shown to express CD4 and low levels of CCR5 and CXCR4, but only myDC express DC SIGN, a C-type lectin that binds human immunodeficiency virus but does not mediate virus entry. Both DC types were more susceptible to infection with a macrophage than a lymphotropic strain of human immunodeficiency virus type 1, but pcDC were more readily infected than myDC. PMID- 11413341 TI - Viral protein U (Vpu)-mediated enhancement of human immunodeficiency virus type 1 particle release depends on the rate of cellular proliferation. AB - Viral protein U (Vpu) is a 17-kDa phosphoprotein that enhances the release of viral particles from human immunodeficiency virus type 1-infected cells. This study shows that the effect of Vpu on efficient particle release depends on the rate of cell proliferation. Cells arrested by contact inhibition, chemical arresting agents, or terminal differentiation (i.e., macrophages) all exhibited a striking dependence on Vpu for efficient particle release, as shown by examination of particle production from transfections with full-length clones, infections, and the vaccinia virus expression system. In contrast, actively proliferating cells did not exhibit enhanced particle release with Vpu expression. This study demonstrates the necessity of Vpu for efficient viral particle release from quiescent cells. PMID- 11413342 TI - Essential role of cyclization sequences in flavivirus RNA replication. AB - A possible role in RNA replication for interactions between conserved complementary (cyclization) sequences in the 5'- and 3'-terminal regions of Flavivirus RNA was previously suggested but never tested in vivo. Using the M fold program for RNA secondary-structure predictions, we examined for the first time the base-pairing interactions between the covalently linked 5' genomic region (first ~160 nucleotides) and the 3' untranslated region (last ~115 nucleotides) for a range of mosquito-borne Flavivirus species. Base-pairing occurred as predicted for the previously proposed conserved cyclization sequences. In order to obtain experimental evidence of the predicted interactions, the putative cyclization sequences (5' or 3') in the replicon RNA of the mosquito-borne Kunjin virus were mutated either separately, to destroy base-pairing, or simultaneously, to restore the complementarity. None of the RNAs with separate mutations in only the 5' or only the 3' cyclization sequences was able to replicate after transfection into BHK cells, while replicon RNA with simultaneous compensatory mutations in both cyclization sequences was replication competent. This was detected by immunofluorescence for expression of the major nonstructural protein NS3 and by Northern blot analysis for amplification and accumulation of replicon RNA. We then used the M-fold program to analyze RNA secondary structure of the covalently linked 5'- and 3'-terminal regions of three tick-borne virus species and identified a previously undescribed additional pair of conserved complementary sequences in locations similar to those of the mosquito-borne species. They base-paired with DeltaG values of approximately -20 kcal, equivalent or greater in stability than those calculated for the originally proposed cyclization sequences. The results show that the base-pairing between 5' and 3' complementary sequences, rather than the nucleotide sequence per se, is essential for the replication of mosquito-borne Kunjin virus RNA and that more than one pair of cyclization sequences might be involved in the replication of the tick-borne Flavivirus species. PMID- 11413343 TI - Population genetic analysis of the protease locus of human immunodeficiency virus type 1 quasispecies undergoing drug selection, using a denaturing gradient heteroduplex tracking assay. AB - Monitoring the evolution of human immunodeficiency virus type 1 (HIV-1) drug resistance requires measuring the frequency of closely related genetic variants making up the complex viral quasispecies found in vivo. In order to resolve both major and minor (>/=2%) protease gene variants differing by one or more nucleotide substitutions, we analyzed PCR products derived from plasma viral quasispecies by using a combination of denaturing gradient gel electrophoresis and DNA heteroduplex tracking assays. Correct population sampling of the high level of genetic diversity present within viral quasispecies could be documented by parallel analysis of duplicate, independently generated PCR products. The composition of genetically complex protease gene quasispecies remained constant over short periods of time in the absence of treatment and while plasma viremia fell >100-fold following the initiation of protease inhibitor ritonavir monotherapy. Within a month of initiating therapy, a strong reduction in the genetic diversity of plasma viral populations at the selected protease locus was associated with rising plasma viremia and the emergence of drug resistance. The high levels of protease genetic diversity seen before treatment reemerged only months later. In one patient, reduction in genetic diversity at the protease gene was observed concomitantly with an increase in diversity at the envelope gene (E. L. Delwart, P. Heng, A. Neumann, and M. Markowitz, J. Virol. 72:2416-2421, 1998), indicating that opposite population genetic changes can take place in different HIV-1 loci. The rapid emergence of drug-resistant HIV-1 was therefore associated with a strong, although only transient, reduction in genetic diversity at the selected locus. The denaturing gradient-heteroduplex tracking assay is a simple method for the separation and quantitation of very closely related, low frequency, genetic variants within complex viral populations. PMID- 11413345 TI - Serious motor vehicle crashes: the cost of untreated sleep apnoea. PMID- 11413346 TI - Revisiting interactions between hypoxaemia and beta2 agonists in asthma. PMID- 11413347 TI - Reduction in motor vehicle collisions following treatment of sleep apnoea with nasal CPAP. AB - BACKGROUND: Patients with untreated obstructive sleep apnoea (OSA) have increased motor vehicle collisions (MVCs). When successfully treated, they report improved driving and fewer mishaps, but there are few objective data to confirm this. A study was therefore undertaken to examine actual MVC data in a large group of patients with OSA before and after treatment with continuous positive airway pressure (CPAP) compared with a control group matched for age, sex, and type of driver's licence (commercial or non-commercial). METHODS: Two hundred and ten patients of mean (SD) age 52 (11) years, body mass index (BMI) 35.5 (10) kg/m(2), apnoea/hypopnoea index (AHI) 54 (29) events/h were treated with CPAP for at least 3 years. MVC records were obtained from the Ontario Ministry of Transportation (MTO) database for patients and an equal number of randomly selected control drivers. MVC rates were compared for 3 years before and after CPAP therapy for patients and for the corresponding time frames for controls. RESULTS: Untreated patients with OSA had more MVCs than controls (mean (SD) MVCs/driver/year 0.18 (0.29) v 0.06 (0.17), p<0.001). Following CPAP treatment the number of MVCs/driver/year fell to normal (0.06 (0.17)) while, in controls, the MVC rate was unchanged over time (0.06 (0.17) v 0.07 (0.18), p=NS). Thus, the change in MVCs over time between the groups was very significant (change = -0.12 (95% CI 0.17 to -0.06), p<0.001)). The MVC rate in untreated patients (n=27) remained high over time. Driving exposure was not different following CPAP. CONCLUSIONS: The risk of MVCs due to OSA is removed when patients are treated with CPAP. As such, any restrictions on driving because of OSA could be safely removed after treatment. PMID- 11413348 TI - Partners of patients with sleep apnoea/hypopnoea syndrome: effect of CPAP treatment on sleep quality and quality of life. AB - BACKGROUND: The sleep apnoea/hypopnoea syndrome (SAHS) causes snoring, apnoeas, and restlessness during sleep which partners frequently complain about. A study was undertaken to determine the impact on partners of SAHS and of treatment of the patient with continuous positive airway pressure (CPAP). METHODS: Forty nine partners and patients with SAHS booked for CPAP treatment completed in house and validated questionnaires (Pittsburgh sleep quality index, Short Form 36 self reported health status) before the patient started treatment. Twenty three couples in whom the index SAHS patient had no driving problems were recruited to a randomised crossover trial with 1 month limbs of CPAP and placebo capsule. At the end of each limb the partners' sleep was monitored by home polysomnography (PSG) and questionnaires were completed. RESULTS: Before treatment partners frequently reported moderate to severe disturbance from patient snoring/apnoeas/restlessness and had poor sleep quality and self-reported health status. In the crossover study (22 completed) the partners' objective sleep quality did not differ between CPAP and placebo, but they reported benefit from treatment of the patients with CPAP in subjective sleep quality (p=0.05) and disturbance to sleep (p=0.03). The reported change in partners' sleep quality between pre-study and following CPAP treatment correlated positively with CPAP use (r=0.5, p=0.01). CONCLUSIONS: Partners of patients with SAHS have poor sleep quality and self-reported health status but only subjective sleep quality benefits from treatment of the patient with CPAP. PMID- 11413349 TI - Effect of pulmonary rehabilitation on exhaled nitric oxide in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: In patients with mild to moderate chronic obstructive pulmonary disease (COPD) the exercise induced increase in exhaled nitric oxide (eNO) parallels that observed in normal untrained subjects. There is no information on the effects of the level of exercise tolerance on eNO in these patients. The aim of this study was to evaluate the effect of a pulmonary rehabilitation programme including exercise training on eNO in patients with COPD. METHODS: In 14 consecutive male patients with stable COPD of mean (SD) age 64 (9) years and forced expiratory volume in one second (FEV1) 55 (14)% predicted, fractional eNO concentration (FeNO), peak work rate (Wpeak) and oxygen uptake (VO2peak) were assessed at baseline (T-1), at the end of a 1 month run in period (T0), and after an 8 week outpatient multidisciplinary pulmonary rehabilitation programme (T1) including cycloergometer training. RESULTS: FeNO did not significantly differ at T-1 and T0 (mean (SE) 4.3 (0.6) and 4.4 (0.6) ppb, respectively), whereas it rose significantly at T1 to 6.4 (0.7) ppb (p<0.02). Compared with T0, both Wpeak and VO2 were significantly (p<0.05) increased at T1 (mean (SE) Wpeak from 89 (5.6) W to 109 (6.9) W); VO2peak from 1.27 (0.1) l/min to 1.48 (0.1) l/min). A significant correlation was found between baseline FEV1 and the change in FeNO following the rehabilitation programme (r=-0.71; p<0.05) and between changes in FeNO and Wpeak from T0 to T1(r=0.60; p<0.05). CONCLUSIONS: Pulmonary rehabilitation in patients with mild to moderate COPD is associated with an increase in exhaled nitric oxide. PMID- 11413350 TI - Long term effects of non-invasive mechanical ventilation on pulmonary haemodynamics in patients with chronic respiratory failure. AB - BACKGROUND: It is not known whether long term nocturnal mechanical ventilation (NMV) reduces pulmonary hypertension in patients with chronic respiratory failure (CRF). METHODS: Pulmonary haemodynamics, spirometric values, and gas exchange were studied in 33 patients requiring NMV due to CRF (20 with thoracic restriction, 13 with chronic obstructive pulmonary disease (COPD)) at baseline and after 1 year of NMV given in the volume cycled mode. Patients with COPD also received supplemental oxygen. RESULTS: Long term NMV improved gas exchange while lung function remained unchanged. Mean pulmonary artery pressure at rest before NMV was higher in patients with thoracic restriction than in those with COPD (33 (10) mm Hg v 25 (6) mm Hg). After 1 year of NMV mean pulmonary artery pressure decreased in patients with thoracic restriction to 25 (6) mm Hg (mean change -8.5 mm Hg (95% CI -12.6 to -4.3), p<0.01) but did not change significantly in patients with COPD (mean change 2.2 mm Hg (95% CI -0.3 to 4.8)). CONCLUSIONS: Long term NMV in CRF improves pulmonary haemodynamics in patients with thoracic restriction but not in patients with COPD. PMID- 11413351 TI - Decreased bronchodilating effect of salbutamol in relieving methacholine induced moderate to severe bronchoconstriction during high dose treatment with long acting beta2 agonists. AB - BACKGROUND: In vitro the long acting beta2 agonist salmeterol can, in contrast to formoterol, behave as a partial agonist and become a partial antagonist to other beta2 agonists. To study this in vivo, the bronchodilating effect of salbutamol was measured during methacholine induced moderate to severe bronchoconstriction in patients receiving maintenance treatment with high dose long acting beta2 agonists. METHODS: A randomised double blind crossover study was performed in 19 asthmatic patients with mean forced expiratory volume in one second (FEV1) of 88.4% predicted and median concentration of methacholine provoking a fall in FEV1 of 20% or more (PC(20)) of 0.62 mg/ml at entry. One hour after the last dose of 2 weeks of treatment with formoterol (24 microg twice daily by Turbuhaler), salmeterol (100 microg twice daily by Diskhaler), or placebo a methacholine provocation test was performed and continued until there was at least a 30% decrease in FEV1. Salbutamol (50 microg) was administered immediately thereafter, followed by ipratropium bromide (40 microg) after a further 30 minutes. Lung function was monitored for 1 hour after provocation. RESULTS: There was a significant bronchodilating and bronchoprotective effect after 2 weeks of active treatment. The dose of methacholine needed to provoke a fall in FEV1 of > or = 30% was higher after pretreatment with formoterol (2.48 mg) than with salmeterol (1.58 mg) or placebo (0.74 mg). The difference between formoterol and salmeterol was statistically significant: 0.7 doubling dose steps (95% CI 0.1 to 1.2, p=0.016). The immediate bronchodilating effect of subsequently administered salbutamol was significantly impaired after pretreatment with both drugs (p<0.0003 for both). Three minutes after inhaling salbutamol the increase in FEV1 relative to the pre-methacholine baseline was 15.8%, 7.3%, and 5.5% for placebo, formoterol and salmeterol, respectively (equivalent to increases of 26%, 14%, and 12%, respectively, from the lowest FEV1 after methacholine). At 30 minutes significant differences remained, but 1 hour after completing the methacholine challenge FEV1 had returned to baseline values in all three treatment groups. CONCLUSION: Formoterol has a greater intrinsic activity than salmeterol as a bronchoprotective agent, indicating that salmeterol is a partial agonist compared with formoterol in contracted human airways in vivo. Irrespective of this, prior long term treatment with both long acting beta2 agonists reduced the bronchodilating effect of an additional single dose of salbutamol equally, indicating that the development of tolerance or high receptor occupancy overshadowed any possible partial antagonistic activity of salmeterol. Patients on regular treatment with long acting beta2 agonists should be made aware that an additional single dose of a short acting beta2 agonist may become less effective. PMID- 11413352 TI - Effect of gas cooking on lung function in adolescents: modifying role of sex and immunoglobulin E. AB - BACKGROUND: A study was undertaken to investigate the effect of gas cooking on the lung function of adolescents while considering serum IgE level as a possible effect modifier. METHOD: The cross sectional study was performed in 702 subjects aged 11-13 years from primary and secondary schools in Civitavecchia and Viterbo ( Latium region in Central Italy), categorised according to how often they were in the kitchen while the mother cooked (never, sometimes, often). Data were collected by questionnaire and lung function was measured by spirometric tests. Bronchial hyperresponsiveness was evaluated by the methacholine test, atopic status by a skin prick test, and a blood sample was collected to determine serum IgE levels. The results were analysed separately for boys and girls. Multiple regression analysis was performed, taking functional parameters (FEV(1), FEV(1)/FVC, FEF(25-75), FEF(50), FEF(75)) as the dependent variables and age, height, parental smoking, and father's education as independent variables. RESULTS: There was no association between time spent in the kitchen and lung function level in boys, but a reduction in lung function was detected in girls which was statistically significant for FEF(75) (sometimes -10.3%, often -11.1%). After stratifying boys and girls into four groups on the basis of the IgE serum level (below and above the median value of IgE), the reduction in lung function was significant in girls with a high IgE value whereas no significant deleterious effects were evident in girls with a low IgE value or in boys with either a low or high IgE. The results remained substantially unchanged after excluding girls with a response to methacholine below the concentration of 4 mg/ml, asthmatic patients, and those with positive skin prick tests. CONCLUSION: Gas cooking has a harmful effect on the lung function of girls with a high serum level of IgE. We do not know whether serum IgE, a marker of allergic susceptibility, is a simple indicator that an inflammatory process is in progress or whether it is involved in the pathogenesis of injury leading to bronchial obstruction. PMID- 11413353 TI - The usefulness of competitive PCR: airway gene expression of IL-5, IL-4, IL 4delta2, IL-2, and IFNgamma in asthma. AB - BACKGROUND: Asthma has been described as an eosinophilic bronchitis driven by interleukin(IL)-4 and IL-5. The quantification of cytokine mRNA levels in airway samples has been confounded by housekeeping gene expression which differs between and within asthmatics and controls. METHODS: The usefulness of competitive reverse transcriptase-polymerase chain reaction (RT-PCR) that is independent of housekeeping gene expression for quantitating the mRNA for interferon (IFN)gamma, IL-2, IL-5, IL-4 and its receptor antagonist encoding splicing variant IL-4delta2 was determined in a cross sectional study of 45 normal control subjects and 111 with asthma. RESULTS: Atopic controls and atopic asthmatic subjects expressed more IL-5 than non-atopic controls (p<0.02) in bronchoalveolar lavage (BAL) cells, but not in biopsy specimens. IL-5 mRNA expression in BAL cells from asthmatic subjects using inhaled corticosteroids (ICS) was significantly lower than those not receiving ICS (p=0.04). IL-2 mRNA levels differed with steroid use in biopsy specimens but not in BAL cells. IFNgamma, IL-4, and IL-4delta2 mRNA levels did not differ between any groups and were not affected by steroid use. IL 4 and IL-4delta2 mRNA levels were positively correlated (p<0.0001), suggesting coordinated transcription. CONCLUSIONS: While the signal differentiation of competitive PCR in asthma may rival that of in situ hybridisation and immunohistochemistry, the method is expensive and wasteful of material. PMID- 11413354 TI - Localisation of transforming growth factor beta1 and beta3 mRNA transcripts in normal and fibrotic human lung. AB - BACKGROUND: Transforming growth factor beta1 is implicated in the pathogenesis of lung fibrosis. It promotes extracellular matrix accumulation by increasing procollagen synthesis and reducing degradation. TGFbeta1 gene and protein expression increase in experimental lung fibrosis, and TGFbeta1 antibodies attenuate fibrosis in mice. The role of other TGFbeta isoforms is unclear. This study aimed to localise TGFbeta1 and TGFbeta3 gene expression in fibrotic human lung and compare it with that in normal human lung. METHODS: Lung tissue from patients with cryptogenic fibrosing alveolitis and fibrosis associated with systemic sclerosis was examined by in situ hybridisation. Macroscopically normal lung from carcinoma resections was used as control tissue. Digoxigenin labelled riboprobes were synthesised from TGFbeta isoform specific cDNA templates. RESULTS: The digoxigenin labelled riboprobes were sensitive and permitted precise cellular localisation of mRNA transcripts. TGFbeta1 and TGFbeta3 mRNA transcripts were widespread in normal lung and localised to alveolar macrophages and bronchiolar epithelium. TGFbeta1 but not TGFbeta3 mRNA was detected in mesenchymal and endothelial cells. In fibrotic lung tissue mRNA transcripts for both isoforms were also detected in metaplastic type II cells. TGFbeta1 gene expression was enhanced in some patients. TGFbeta3 was expressed in fibrotic lung but was not consistently altered compared with controls. CONCLUSION: TGFbeta1 mRNA transcripts were localised in normal and fibrotic human lung and TGFbeta3 gene expression in human lung fibrosis was shown for the first time. The results suggest that TGFbeta1 may play the predominant role in pathogenesis. It is suggested that TGFbeta1 should be the primary target of anticytokine treatments for pulmonary fibrosis. PMID- 11413355 TI - Induced sputum cell profiles in lung transplant recipients with or without chronic rejection: correlation with lung function. AB - BACKGROUND: Sputum induction is a non-invasive procedure for measuring inflammatory processes of the lower respiratory tract. The aim of this study was to establish sputum cell counts and differentials in patients after lung transplantation (LTx), with or without chronic transplant rejection. METHODS: Sputum induction was performed in 41 LTx patients (25 single LTx (sLTx), 16 double LTx (dLTx) and 15 healthy non-smoking volunteers. Sputum was processed according to standard protocols. Total cell count was calculated as mean (SE) cells x 10(6)/ml sputum and cell differential (%) was evaluated after staining. Cellular profiles were correlated with lung function. RESULTS: Total sputum cell counts were increased in sLTx (9 (1.9) cells x 10(6)/ml, p=0.01) and dLTx patients (7.2 (1.5) x 10(6)/ml, p=0.01) compared with healthy controls (2.6 (0.6) x 10(6)/ml). There was also a marked sputum neutrophilia in both patient groups (59 (6)% and 62 (6)%, respectively, p<0.001 v controls). Moreover, in both sLTx and dLTx patients with chronic transplant rejection there was an increased number of sputum neutrophils compared with patients with normal graft function (p<0.05 both comparisons), and neutrophils were inversely correlated with lung function (forced expiratory volume in one second (FEV(1)) % predicted): sLTx, r=-0.61, p=0.001; dLTx, r=-0.75, p=0.001, respectively). Sputum lymphocytes and eosinophils were similar in both groups. No relevant side effects occurred during sputum induction. CONCLUSIONS: Sputum induction is a safe and non-invasive tool for monitoring lower respiratory tract inflammation in LTx patients. Both sLTx and dLTx patients with chronic rejection had increased sputum neutrophils compared with patients with normal transplant function. These data support findings of other authors highlighting a possible role for neutrophils in the pathogenesis of chronic transplant rejection. PMID- 11413356 TI - A biological staging model for operable non-small cell lung cancer. AB - BACKGROUND: Currently the best prognostic index for operable non-small cell lung cancer (NSCLC) is the TNM staging system. Molecular biology holds the promise of predicting outcome for the individual patient and identifying novel therapeutic targets. Angiogenesis, matrix metalloproteinases (MMP)-2 and -9, and the erb/HER type I tyrosine kinase receptors are all implicated in the pathogenesis of NSCLC. METHODS: A retrospective analysis of 167 patients with resected stage I-IIIa NSCLC and >60 days postoperative survival with a minimum follow up of 2 years was undertaken. Immunohistochemical analysis was performed on paraffin embedded sections for the microvessel marker CD34, MMP-2 and MMP-9, EGFR, and c-erbB-2 to evaluate the relationships between and impact on survival of these molecular markers. RESULTS: Tumour cell MMP-9 (HR 1.91 (1.23-2.97)), a high microvessel count (HR 1.97 (1.28-3.03)), and stage (stage II HR 1.44 (0.87-2.40), stage IIIa HR 2.21 (1.31-3.74)) were independent prognostic factors. Patients with a high microvessel count and tumour cell MMP-9 expression had a worse outcome than cases with only one (HR 1.68 (1.04-2.73)) or neither (HR 4.43 (2.29-8.57)) of these markers. EGFR expression correlated with tumour cell MMP-9 expression (p<0.001). Immunoreactivity for both of these factors within the same tumour was associated with a poor prognosis (HR 2.22 (1.45-3.41)). CONCLUSION: Angiogenesis, EGFR, and MMP-9 expression provide prognostic information independent of TNM stage, allowing a more accurate outcome prediction for the individual patient. The development of novel anti-angiogenic agents, EGFR targeted therapies, and MMP inhibitors suggests that target specific adjuvant treatments may become a therapeutic option in patients with resected NSCLC. PMID- 11413357 TI - Cardiovascular side effects of inhaled salbutamol in hypoxic asthmatic patients. AB - BACKGROUND: Beta-2 adrenoceptor agonists have been associated with sudden death in asthma patients but the cause and underlying mechanism are unclear. Animal experiments indicate that the combination of hypoxia and beta2 agonists may result in detrimental cardiovascular effects. A study was undertaken to investigate the effect of hypoxia on the systemic vascular effects of salbutamol in patients with asthma who are hypoxic by assessing forearm blood flow (FBF) as a measure of peripheral vasodilatation. METHODS: Eight men with mild asthma underwent the following treatments: normoxia + placebo (NP), normoxia + salbutamol (NS), hypoxia + placebo (HP), and hypoxia + salbutamol (HS). The period of mask breathing started at t=0 minutes, lasted for 60 minutes, and at 30 minutes 800 microg salbutamol was inhaled. The experiment was completed 30 minutes after the inhalation (t=60 minutes). For the hypoxia treatment the SpO2 level was 82%. Differences between treatments were sought using factorial ANOVA on percentage change from the pretreatment value. RESULTS: There were no significant differences in blood pressure and potassium levels between the treatments. After 60 minutes the increase in FBF was 13% (95% CI -12 to 39) more for HP treatment than for NP, 21% (95% CI -5 to 46) more for NS than for NP, and 32% (95% CI 7 to 58) more for HS than for HP (p=0.016). The inhalation of salbutamol during hypoxia resulted in a significant increase in FBF of 45% (95% CI 20 to 71) compared with NP (p=0.001). CONCLUSION: Patients with asthma who are hypoxic and inhale beta2 agonists have serious systemic vascular side effects which may be an additional explanation for the association between asthma treatment and sudden death. PMID- 11413358 TI - Prevalence of asthma among schoolchildren in Patras, Greece: three surveys over 20 years. AB - BACKGROUND: The aim of the present study was to compare the prevalence of asthma among schoolchildren in 1978, 1991, and 1998 in Patras, Greece. METHODS: The study populations of the three comparable cross sectional surveys comprised third and fourth grade public school children in Patras, Greece. Sample sizes in 1978, 1991, and 1998 were 3735, 2952 and 3397 children and response rates were 80.4%, 81.9%, and 90.6%, respectively. Prevalence of current, non-current, and lifetime asthma or recurrent wheezing was determined by parental questionnaire. Personal communication with the parents of asthmatic children in 1991 and 1998 provided data on lost schooldays. RESULTS: Prevalence rates of current asthma or wheezing in 1978, 1991, and 1998 were 1.5%, 4.6%, and 6.0%, respectively (1978-91: p=0.01, 1991-98: p=0.02, 1978-98: p=0.03). Lifetime prevalences of asthma or wheezing in 1991 and 1998 were 8.0% and 9.6%, respectively (p=0.03). Current diagnosed asthma increased proportionally to diagnosed wheezing during 1991-98. The number of schooldays lost in the previous 2 years because of asthma did not change (p>0.1) between 1991 (0.31 per child) and 1998 (0.34 per child). CONCLUSIONS: Our results support a true increase in the prevalence of current and lifetime asthma in the last 20 years among pre-adolescent children in Patras, Greece. PMID- 11413359 TI - Pulmonary manifestations of Behcet's disease. PMID- 11413360 TI - Smoking cessation: integrating recent advances into clinical practice. AB - Smoking remains a prevalent habit with serious consequences for public health. There are now effective treatments for nicotine addiction and, in the UK, specialist services for the treatment of smoking cessation are becoming available in all areas. This paper reviews the role of treatments for nicotine addiction in the management of smoking cessation. Recommendations are made for the judicious use of these therapies and also for the rational use of the new UK smoking cessation services. PMID- 11413361 TI - Intramural neurofibroma of the trachea treated by multiple stents. AB - The case history is presented of a patient in whom an intramural tracheal neurofibroma developed, causing severe airway stenosis. The patient was treated with multiple stents over a period of 5 years because of progression of the disease and associated airflow limitation. Clinicians should be aware of this rare complication of neurofibromatosis. PMID- 11413363 TI - Evidence for structural differences in the S domain of L in comparison with S protein of hepatitis B virus. AB - The structures of the large (L), middle (M) and small (S) versions of the envelope proteins of hepatitis B virus remain poorly characterized due to the complex nature of their conformations. Several groups have proposed transmembrane topological models depicting the lumenally and cytosolically disposed regions of these proteins. Recently, post-translational topological changes in L have been described. However, no overall differences in the topology of the S domains of the L or M, to the S protein are predicted. In this report, we investigated a previously uncharacterized anti-S monoclonal antibody (MAb), 6B1, which recognizes a conformation-sensitive epitope in S. Unlike other anti-S MAbs tested, this MAb did not recognize its epitope in the S domain of L protein. Interestingly, however, the M protein was efficiently recognized. This unique characteristic of MAb 6B1 has allowed us to study the intracellular distribution of L and S proteins. In cells expressing both L and S, L re-localized from the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) to the membrane associated distribution of S protein indicating that L and S interact with each other. This was confirmed by immunoprecipitation assays, which also showed that the interaction between L and S results in the secretion of L protein from cells. Overall, the ability of MAb 6B1 to selectively recognize S and M, but not L, strongly points to the existence of significant topological differences in the S domain of L. The availability of this important reagent should help further our understanding of the structure of HBV surface antigens. PMID- 11413364 TI - Coordinate effects of human immunodeficiency virus type 1 protein Tat and cellular protein Puralpha on DNA replication initiated at the JC virus origin. AB - JC virus (JCV) causes progressive multifocal leukoencephalopathy, a demyelinating disease in brains of individuals with AIDS. Previous work has shown that the Tat protein, encoded by human immunodeficiency virus type 1 (HIV-1), can interact with cellular protein Puralpha to enhance both TAR-dependent HIV-1 transcription and JCV late gene transcription. Tat has been shown to activate JCV transcription through interaction with Puralpha, which binds to promoter sequence elements near the JCV origin of replication. DNA footprinting has shown that Puralpha and large T-antigen cooperatively interact at several binding sites in the origin and transcriptional control region. Overexpression of Puralpha inhibits replication initiated at the JCV origin by T-antigen. In transfected glial cells Tat reversed this inhibition and enhanced DNA replication. In an in vitro replication system maximal activation by Tat, more than sixfold the levels achieved with T-antigen alone, was achieved in the presence of Puralpha. Effects of mutant Tat proteins on both activation of replication and binding to Puralpha have revealed that Cys22 exerts a conformational effect that affects both activities. The origin of an archetypal strain of JCV was less susceptible to activation of replication by Tat relative to the rearranged Mad-1 strain. These results have revealed a previously undocumented role for Tat in DNA replication and have indicated a regulatory role for JCV origin auxiliary sequences in replication and activation by Tat. PMID- 11413365 TI - Analysis of canine parvovirus sequences from wolves and dogs isolated in Italy. AB - The VP2 genes of Italian canine parvovirus (CPV) type 2 strains isolated from dogs and wolves were sequenced and a three-dimensional model of the VP2 capsid protein was constructed. Two mutations were detected in the VP2 sequences of the Italian strains: one at residue 297 and one at residue 265. Variant 297 is the predominant CPV isolate in Europe, whereas variant 265 has never been detected before. The mutation at residue 265 causes a disruption in a G strand of the beta barrel in the VP2 protein. Data on strains isolated from wolves demonstrated that the same strain of CPV can circulate among domestic and wild canids; therefore, this result leads us to exclude the possibility that a separate parvovirus pool exists in wild populations. PMID- 11413366 TI - Evaluation of the primary effect of brefeldin A treatment upon herpes simplex virus assembly. AB - Addition of the drug brefeldin A (BFA) to cells infected by herpes simplex virus (HSV) type 1 is known to result in a complex pattern of defects in particle assembly. BFA-treated, infected cells accumulate perinuclear enveloped virions and non-enveloped ("naked") cytoplasmic capsids, and it has been difficult to interpret these data in terms of the assembly pathway of HSV and the known effects of BFA on the secretory apparatus. Since BFA is a cytotoxic drug, and earlier studies commonly examined the effects of long-term BFA incubations on infected cells, it was hypothesized that the drug could have pleiotropic and indirect effects on HSV assembly. To test this, use was made of an HSV synchronized assembly assay, in which cells are infected with the virus mutant tsProt.A and maintained at 39 degrees C to induce reversible accumulation of a population of procapsids. By first adding BFA and then shifting these cells to 31 degrees C for 3 h to allow the accumulated procapsids to mature, it was possible to test the effect of short-term BFA treatment on only those HSV assembly events that are downstream of procapsid maturation. Under these conditions, it was found that procapsids matured and packaged the viral genome normally, but remained non enveloped and failed to exit the nucleus. It is concluded that the primary effect of BFA on HSV replication is to inhibit budding at the inner nuclear membrane. PMID- 11413367 TI - The carboxyl terminus of the human cytomegalovirus UL37 immediate-early glycoprotein is conserved in primary strains and is important for transactivation. AB - The human cytomegalovirus (HCMV) UL37 exon 3 (UL37x3) open reading frame (ORF) encodes the carboxyl termini of two immediate-early glycoproteins (gpUL37 and gpUL37(M)). UL37x3 homologous sequences are not required for mouse cytomegalovirus (MCMV) growth in vitro; yet, they are important for MCMV growth and pathogenesis in vivo. Similarly, UL37x3 sequences are dispensable for HCMV growth in culture, but their requirement for HCMV growth in vivo is not known. To determine this requirement, we directly sequenced the complete UL37x3 gene in multiple HCMV primary strains. A total of 63 of the 310 amino acids in the UL37x3 ORF differ non-conservatively in one or more HCMV primary strains. The HCMV UL37x3 genetic diversity is non-random: the N-glycosylation (46/186 aa) and basic (9/15 aa) domains have the highest proportion of non-conservative variant amino acids. Nonetheless, most (15/17 signals) of the N-glycosylation signals are retained in all HCMV primary strains. Moreover, new N-glycosylation signals are encoded by 5/20 primary strains. In sharp contrast, the UL37x3 transmembrane (TM) ORF completely lacks diversity in all 20 HCMV sequenced primary strains, and only 1 of 28 cytosolic tail residues differs non-conservatively. To test the functional significance of the conserved carboxyl terminus, gpUL37 mutants lacking the TM and/or cytosolic tail were tested for transactivating activity. The gpUL37 carboxyl-terminal mutants are partially defective in hsp70 promoter transactivation even though they trafficked similarly to the wild-type protein into the endoplasmic reticulum and to mitochondria. From these results, we conclude that N-glycosylated gpUL37, particularly its TM and cytosolic domains, is important for HCMV growth in humans. PMID- 11413368 TI - Epstein-Barr virus nuclear antigen 5 interacts with HAX-1, a possible component of the B-cell receptor signalling pathway. AB - Using a yeast two-hybrid screen of a B-cell cDNA library with an Epstein-Barr nuclear antigen 5 (EBNA5) molecule containing seven repeats of the W(1)W(2) domain as bait, we have isolated the EBNA5-interacting protein HAX-1. HAX-1 has previously been shown to associate with HS1, a protein specifically expressed in cells of the haematopoietic lineage, and is thought to be involved in signal transduction in B-cells. Immunofluorescence experiments showed that HAX-1 co localized with the hsp60 protein that is associated with the mitochondria in the cell cytoplasm. Pull down experiments with a fusion protein between glutathione S transferase and the seven copy repeat EBNA5 synthesized in bacteria and in yeast cells confirmed that HAX-1 can interact with EBNA5 in vitro. Conventionally, EBNA5 is regarded as a nuclear protein. However, we show here that the smallest EBNA5 species, composed of the unique Y domain and only one copy of the W(1)W(2) repeat domain, like HAX-1, co-localizes with the mitochondrial hsp60 protein in the B-cell cytoplasm. Furthermore, immunoprecipitation experiments demonstrate that the single repeat EBNA5 associates with HAX-1 in transfected B lymphoblastoid cells. PMID- 11413369 TI - Feline leukaemia provirus load during the course of experimental infection and in naturally infected cats. AB - Feline leukaemia virus (FeLV) infection in domestic cats can vary in its outcome (persistent, transient, no infection) for reasons that are not entirely known. It was hypothesized that the initial virus and provirus load could significantly influence the course of retrovirus infection. To determine the role of provirus loads, two methods of PCR, a nested PCR and a fluorogenic probe-based (TaqMan) real-time quantitative PCR, which were specific to the U3 region of FeLV-A were established. FeLV provirus in naturally and experimentally infected cats was then measured. Only 3 weeks after experimental FeLV-A infection, persistently infected cats demonstrated higher provirus loads and lower humoral immune responses than cats that had overcome antigenaemia. Lower initial provirus loads were associated with successful humoral immune responses. Unexpectedly, provirus in the buffy coat cells of two cats that tested negative for the p27 antigen (a marker for viraemia) was also detected. In 597 Swiss cats, comparison of p27 antigen levels with PCR results revealed broad agreement. However, similar to the experimental situation, a significant number of animals (10%) was negative for the p27 antigen and FeLV-positive by PCR. These cats had a mean provirus load 300-fold lower than that of animals testing positive for the p27 antigen. In conclusion, an association between the provirus load and the outcome of FeLV infection was found. Detection of provirus carriers should contribute to further the control of FeLV. In addition, quantification of provirus loads will lead to a better understanding of FeLV pathogenesis and anti-retrovirus protective mechanisms. PMID- 11413370 TI - The envelope of Mason-Pfizer monkey virus has immunosuppressive properties. AB - We have demonstrated previously that the envelope protein of a murine retrovirus, Moloney murine leukaemia virus, has immunosuppressive properties in vivo. This property was manifested by the ability of the protein, when expressed by tumour cells normally rejected by engrafted mice, to allow the env-expressing cells to escape immune rejection and to proliferate. Here, it is shown that this property is not restricted to the envelope of a murine retrovirus, but is also shared by the envelope encoded by a primate retrovirus, Mason-Pfizer monkey virus. PMID- 11413371 TI - Cross-protection against mucosal simian immunodeficiency virus (SIVsm) challenge in human immunodeficiency virus type 2-vaccinated cynomolgus monkeys. AB - In this study we compared the efficacy of live attenuated human immunodeficiency virus type 2 (HIV-2) vaccine alone versus boosting with live non-pathogenic HIV-2 following priming with ALVAC HIV-2 (recombinant canarypox virus expressing HIV-2 env, gag and pol). Six monkeys were first inoculated intravenously with live HIV 2(SBL-6669) and 7 to 10 months later were challenged intrarectally with 10 MID(50) of cell-free simian immunodeficiency virus (SIV) strain SIVsm. One monkey was completely protected against SIV infection and all five monkeys that became SIV-infected showed a lower virus replication and an initial lower virus load as compared with a parallel group of six control animals. In another experiment five monkeys were immunized either three times with ALVAC HIV-2 alone or twice with ALVAC HIV-2 and once with purified native HIV-2 gp125. The monkeys were then challenged with HIV-2 given intravenously and finally with pathogenic SIVsm given intrarectally. After challenge with SIVsm, three of five monkeys were completely protected against SIVsm infection whereas the remaining two macaques became SIV infected but with limited virus replication. In conclusion, vaccination with an ALVAC HIV-2 vaccine followed by exposure to live HIV-2 could induce cross protection against mucosal infection with SIVsm and seemed to be more efficient than immunization with a live HIV-2 vaccine only. PMID- 11413372 TI - Insertion/deletion frequencies match those of point mutations in the hypervariable regions of the simian immunodeficiency virus surface envelope gene. AB - A method for encoding insertions and deletions (indels) has been developed and adapted to the SplitsTree program. Following phylogenetic reconstruction, the relative frequencies of indels were estimated for a large number of in vivo sequence sets corresponding to the env V1 hypervariable region of the simian immunodeficiency virus SIVmac251. The method allowed recovery of many point mutations hitherto lost due to gap stripping. Deletions were as frequent as transversions and were 4- to 8-fold more frequent than insertions, invariably duplications. The high proportion of deletions among mutation events suggests that lentivirus vectors may readily delete parts of their cargo. PMID- 11413373 TI - Phylogenetic reconstruction of intrapatient evolution of human immunodeficiency virus type 1: predominance of drift and purifying selection. AB - The intra-host evolution of 73 human immunodeficiency virus type 1 quasispecies was analysed by split decomposition analysis. Non-synonymous and synonymous nucleotide substitutions were counted along the shortest path connecting all sequences and compared with the numbers expected under the assumption of a random model of mutation. For the majority of substitutions, drift and negative selection seemed to prevail. PMID- 11413374 TI - Coexpression of hepatitis C virus envelope proteins E1 and E2 in cis improves the stability of membrane insertion of E2. AB - The hepatitis C virus (HCV) genome encodes two envelope glycoproteins, E1 and E2. These proteins contain a large N-terminal ectodomain, and are anchored into membranes by their C-terminal transmembrane domain (TMD). The TMDs of HCV envelope proteins are multifunctional. In addition to their role as membrane anchors, they possess a signal sequence function in their C-terminal half, and play a major role in subcellular localization and assembly of these envelope proteins. In this work, the expression of full-length E2 led to secretion of a proportion of this protein, which is likely to be due to inefficient membrane insertion of a fraction of E2 expressed alone. However, when E1 and E2 were coexpressed from the same polyprotein, E2 was not secreted and remained tightly associated with membranes, suggesting that an early interaction between the TMDs of HCV envelope proteins improves the stability of membrane insertion of E2. These results reinforce the hypothesis that the TMDs of E1 and E2 are major factors in the assembly of the HCV envelope glycoprotein complex. PMID- 11413375 TI - The inhibition of cAMP-dependent protein kinase by full-length hepatitis C virus NS3/4A complex is due to ATP hydrolysis. AB - Hepatitis C virus (HCV) is an important cause of chronic liver disease, but the molecular mechanisms of viral pathogenesis remain to be established. The HCV non structural protein NS3 complexes with NS4A and has three enzymatic activities: a proteinase and a helicase/NTPase. Recently, catalytically inactive NS3 fragments containing an arginine-rich motif have been reported to interact with, and inhibit, the catalytic subunit of cAMP-dependent protein kinase (PKA C-subunit). Here we demonstrate that full-length, catalytically active NS3/4A, purified from recombinant baculovirus-infected insect cells, is also able to inhibit PKA C subunit in vitro. This inhibition was abrogated by mutation of either the arginine-rich motif or the conserved helicase motif II, both of which also abolished NTPase activity. As PKA C-subunit inhibition was also enhanced by poly(U) (an activator of NS3 NTPase activity), we hypothesized that PKA C-subunit inhibition could be due to NS3/4A-mediated ATP hydrolysis. This was confirmed by experiments in which a constant ATP concentration was maintained by addition of an ATP regeneration system--under these conditions PKA C-subunit inhibition was not observed. Interestingly, the mutations also abrogated the ability of wild type NS3/4A to inhibit the PKA-regulated transcription factor CREB in transiently transfected hepatoma cells. Our data are thus not consistent with the previously proposed model in which the arginine-rich motif of NS3 was suggested to act as a pseudosubstrate inhibitor of PKA C-subunit. However, in vivo effects of NS3/4A suggest that ATPase activity may play a role in viral pathology in the infected liver. PMID- 11413376 TI - Mutagenesis of the dengue virus type 2 NS3 proteinase and the production of growth-restricted virus. AB - The N-terminal one-third of the NS3 protein of Dengue virus type 2 (DEN-2) complexes with co-factor NS2B to form an active serine proteinase which cleaves the viral polyprotein. To identify sites within NS3 that may interact with NS2B, seven regions within the NS3 proteinase outside the conserved flavivirus enzyme motifs were mutated by alanine replacement. Five sites contained clusters of charged residues and were hydrophilic. Two sites were hydrophobic and highly conserved among flaviviruses. The effects of five mutations on NS2B/3 processing were examined using a COS cell expression system. Four retained significant proteinase activity. Three of these mutations and two more were introduced into genomic-length cDNA and tested for their effects on virus replication. The five mutant viruses showed reduced plaque size and two of the five showed significantly reduced titres. All seven mutations were mapped on the X-ray crystal structure of the DEN-2 NS3 proteinase: three were located at the N terminus and two at the C terminus of the NS2B-binding cleft. Two mutations were at the C terminus of the proteinase domain and one was solvent-exposed. The study demonstrated that charged-to-alanine mutagenesis in the viral proteinase can be used to produce growth-restricted flaviviruses that may be useful in the production of attenuated vaccine strains. PMID- 11413377 TI - Phylogeny of North American Powassan virus. AB - To determine whether Powassan virus (POW) and deer tick virus (DTV) constitute distinct flaviviral populations transmitted by ixodid ticks in North America, we analysed diverse nucleotide sequences from 16 strains of these viruses. Two distinct genetic lineages are evident, which may be defined by geographical and host associations. The nucleotide and amino acid sequences of lineage one (comprising New York and Canadian POW isolates) are highly conserved across time and space, but those of lineage two (comprising isolates from deer ticks and a fox) are more variable. The divergence between lineages is much greater than the variation within either lineage, and lineage two appears to be more diverse genetically than is lineage one. Application of McDonald-Kreitman tests to the sequences of these strains indicates that adaptive evolution of the envelope protein separates lineage one from lineage two. The two POW lineages circulating in North America possess a pattern of genetic diversity suggesting that they comprise distinct subtypes that may perpetuate in separate enzootic cycles. PMID- 11413378 TI - The degree of attenuation of tick-borne encephalitis virus depends on the cumulative effects of point mutations. AB - An infectious clone (pGGVs) of the tick-borne encephalitis complex virus Vasilchenko (Vs) was constructed previously. Virus recovered from pGGVs produced slightly smaller plaques than the Vs parental virus. Sequence analysis demonstrated five nucleotide differences between the original Vs virus and pGGVs; four of these mutations resulted in amino acid substitutions, while the fifth mutation was located in the 3' untranslated region (3'UTR). Two mutations were located in conserved regions and three mutations were located in variable regions of the virus genome. Reverse substitutions from the conserved regions of the genome, R(496)-->H in the envelope (E) gene and C(10884)-->T in the 3'UTR, were introduced both separately and together into the infectious clone and their biological effect on virus phenotype was evaluated. The engineered viruses with R(496) in the E protein produced plaques of smaller size than viruses with H(496) at this position. This mutation also affected the growth and neuroinvasiveness of the virus. In contrast, the consequence of a T(10884)-->C substitution within the 3'UTR was noticeable only in cytotoxicity and neuroinvasiveness tests. However, all virus mutants engineered by modification of the infectious clone, including one with two wild-type mutations, H(496) and T(10884), showed reduced neuroinvasiveness in comparison with the Vs parental virus. Therefore, although the H(496)-->R and T(10884)-->C substitutions clearly reduce virus virulence, the other mutations within the variable regions of the capsid (I(45)-->F) and the NS5 (T(2688)-->A and M(3385)-->I) genes also contribute to the process of attenuation. In terms of developing flavivirus vaccines, the impact of accumulating apparently minor mutations should be assessed in detail. PMID- 11413379 TI - Antigen-presenting cells from calves persistently infected with bovine viral diarrhoea virus, a member of the Flaviviridae, are not compromised in their ability to present viral antigen. AB - The aim of this study was to assess whether the infection of antigen-presenting cells (APC) in vivo, evident in calves persistently infected (PI) with bovine viral diarrhoea virus (BVDV), compromised their ability to stimulate virus specific T cell responses. Major histocompatibility complex (MHC) molecule identical cattle were identified from the inbred family at the Institute for Animal Health. One was PI and immunotolerant to BVDV. Virus was not isolated from the remaining calves, which were classified as BVDV-immune or BVDV-naive depending on the presence or absence of BVDV-specific antibodies in sera. Two colour flow-cytometric analysis of PBMC from the PI calf showed that 40% of CD14(+) monocytes were infected in vivo. Monocytes from the PI calf (PI monocytes) were used as naturally infected ex vivo APC with CD4(+) or CD8(+) T cells isolated from the BVDV-naive or BVDV-immune animals. PI monocytes stimulated proliferative responses with CD4(+) and CD8(+) T cells from BVDV immune animals, but not from BVDV-naive calves. This provided evidence for the presence of virus-specific CD4(+) and CD8(+) memory T cells after acute infection and indicated that ex vivo monocytes from PI, immunotolerant calves stimulated both MHC class I- and MHC class II-restricted T cell responses to BVDV. Additionally, naturally infected ex vivo monocytes cultured in vitro for 3 days stimulated effective T cell responses to the virus with which they were infected. PMID- 11413380 TI - Acute hepatitis caused by a novel strain of hepatitis E virus most closely related to United States strains. AB - A unique hepatitis E virus (HEV) strain was identified as the aetiological agent of acute hepatitis in a United States (US) patient who had recently returned from vacation in Thailand, a country in which HEV is endemic. Sequence comparison showed that this HEV strain was most similar, but not identical, to the swine and human HEV strains recovered in the US. Phylogenetic analysis revealed that this new HEV isolate was closer to genotype 3 strains than to the genotype 1 strains common in Asia. The fact that this HEV was closely related to strains recovered in countries where HEV is not endemic and was highly divergent from Asian HEV strains raises the questions of where the patient's infection was acquired and of whether strains are geographically as localized as once thought. PMID- 11413381 TI - Identification of conformational neutralizing epitopes on the capsid protein of canine calicivirus. AB - Two neutralizing monoclonal antibodies (MAbs) against canine calicivirus (CaCV), which has a distinct antigenicity from feline calicivirus (FCV), were obtained. Both MAbs recognized conformational epitopes on the capsid protein of CaCV and were used to identify these epitopes. Neutralization-resistant variants of CaCV were selected in the presence of individual MAbs in a cell culture. Cross neutralization tests using the variants indicated that the MAbs recognized functionally independent epitopes on the capsid protein. Recombinantly expressed ORF2 products (capsid precursors) of the variants showed no reactivity to the MAbs used for the selection, suggesting that the resistance was induced by a failing in binding of the MAbs to the variant capsid proteins. Several nucleotide changes resulting in amino acid substitutions in the capsid protein were found by sequence analysis. Reactivities of the MAbs to the revertant ORF2 products produced from each variant ORF2 by site-directed mutagenesis identified a single amino acid substitution in each variant capsid protein responsible for the failure of MAb binding. The amino acid residues related to forming the conformational neutralizing epitopes were located in regions equivalent to the 5' and 3' hypervariable regions of the FCV capsid protein, where antigenic sites were demonstrated in previous studies. The recombinant ORF2 products expressed in bacteria failed to induce neutralizing antibody, suggesting that neutralizing antibodies were only generated when properly folded capsid protein was used as an antigen. In CaCV, the conformational epitopes may play a more important role in neutralization than do linear epitopes. PMID- 11413382 TI - Foot-and-mouth disease virus can utilize the C-terminal extension of coxsackievirus A9 VP1 for cell infection. AB - Foot-and-mouth disease virus (FMDV) is known to employ the conserved Arg-Gly-Asp (RGD) tripeptide located on the variable betaG-betaH loop of the VP1 capsid protein for binding to cells. Coxsackievirus A9 (CAV9) also carries an RGD sequence, but on a short C-terminal extension of its VP1 and in a different amino acid context. This apparent relationship raised the question of whether insertion of the heterologous CAV9 sequence into FMDV would influence infection by the genetically modified FMDV. Four VP1 mutants were generated by PCR mutagenesis of a full-length FMDV cDNA plasmid. After transfection of BHK-21 cells, viral protein synthesis and virus particle formation could be detected. Two of the four mutants, mV9b and mV9d, could be propagated in BHK-21 cells, but not in CV-1 cells. Both of these mutants contained 17 amino acids of the C terminus of CAV9 VP1. Infection of BHK cells could be specifically inhibited by rabbit immune serum raised against a synthetic peptide representing the amino acid sequence of the C-terminal extension of CAV9 VP1. This demonstrated the direct involvement of the inserted sequence in cell infection. In fact, genetically modified FMDV O(1)K was capable of employing the VP1 C-terminal RGD region of CAV9 for infection of BHK cells. In addition, these results show that, even in cell culture-adapted viruses, the RGD-containing betaG-betaH loop plays an important role in virus infectivity. PMID- 11413383 TI - Induction of a protective response in swine vaccinated with DNA encoding foot-and mouth disease virus empty capsid proteins and the 3D RNA polymerase. AB - This work focuses on the development of a potential recombinant DNA vaccine against foot-and-mouth disease virus (FMDV). Such a vaccine would have significant advantages over the conventional inactivated virus vaccine, in particular having none of the risks associated with the high security requirements for working with live virus. The principal aim of this strategy was to stimulate an antibody response to native, neutralizing epitopes of empty FMDV capsids generated in vivo. Thus, a plasmid (pcDNA3.1/P1-2A3C3D) was constructed containing FMDV cDNA sequences encoding the viral structural protein precursor P1 2A and the non-structural proteins 3C and 3D. The 3C protein was included to ensure cleavage of the P1-2A precursor to VP0, VP1 and VP3, the components of self-assembling empty capsids. The non-structural protein 3D was also included in the construct in order to provide additional stimulation of CD4(+) T cells. When swine were immunized with this plasmid, antibodies to FMDV and the 3D polymerase were synthesized. Furthermore, neutralizing antibodies were detected and, after three sequential vaccinations with DNA, some of the animals were protected against challenge with live virus. Additional experiments suggested that the antibody response to FMDV proteins was improved by the co-administration of a plasmid encoding porcine granulocyte-macrophage colony-stimulating factor. Although still not as effective as the conventional virus vaccine, the results encourage further work towards the development of a DNA vaccine against FMDV. PMID- 11413384 TI - Equine rhinitis A virus: structural proteins and immune response. AB - Equine rhinitis A virus (ERAV) is a picornavirus that has been reclassified as a member of the Aphthovirus genus because of its resemblance to foot-and-mouth disease virus at the level of nucleotide sequence and overall genomic structure. The N-terminal amino acid sequence of three of the four capsid proteins of ERAV was determined and showed that the proteolytic cleavage sites within the precursor P1 polypeptide occur exactly as those predicted for an aphthovirus-like 3C protease, which generates the capsid proteins VP1 and VP3. However, the autocatalytic cleavage site between VP4 and VP2, which is independent of 3C protease cleavage, was different from that predicted previously. ERAV.393/76 antisera from horses and rabbits showed different reactivity to the viral structural proteins in both serum neutralization assays and Western blots. High neutralizing antibody titres appeared to correlate with strong reactivity to VP1 in Western blots. PMID- 11413385 TI - High-level expression of a foreign gene from the most 3'-proximal locus of a recombinant Newcastle disease virus. AB - A previous report showed that insertion of a foreign gene encoding chloramphenicol acetyltransferase (CAT) between the HN and L genes of the full length cDNA of a virulent Newcastle disease virus (NDV) yielded virus with growth retardation and attenuation. The NDV vector used in that study was pathogenic to chickens; it is therefore not suitable for use as a vaccine vector. In the present study, an avirulent NDV vector was generated and its potential to express CAT protein was evaluated. The CAT gene was under the control of NDV transcriptional start and stop signals and was inserted immediately before the open reading frame of the viral 3'-proximal nucleocapsid protein gene. A recombinant NDV expressing CAT activity at a high level was recovered. The replication and pathogenesis of the CAT-expressing recombinant NDV were not modified significantly. These results indicate the potential utility of an avirulent NDV as a vaccine vector. PMID- 11413386 TI - Naked RNA immunization with replicons derived from poliovirus and Semliki Forest virus genomes for the generation of a cytotoxic T cell response against the influenza A virus nucleoprotein. AB - The potential of RNA-based vaccines was evaluated for the generation of a protective immune response in the mouse model of influenza type A virus infection using the internal nucleoprotein (NP) as antigen. This antigen is of particular interest, since it has the potential to elicit protective cytotoxic T lymphocytes (CTL) against heterologous strains of influenza A virus. In view of the short half-life of RNA, self-replicating RNAs or replicons of the positive-stranded genomes of Semliki Forest virus (SFV) and poliovirus were engineered to synthesize the influenza A virus NP in place of their structural proteins. NP expression was demonstrated by immunoprecipitation after transfection of cells with RNA from the SFV (rSFV-NP) and poliovirus (rDeltaP1-E-NP) genome-derived replicons transcribed in vitro. C57BL/6 mice were injected intramuscularly with these synthetic RNAs in naked form. Both replicons, rSFV-NP and rDeltaP1-E-NP, induced antibodies against the influenza virus NP, but only mice immunized with the rSFV-NP replicon developed a CTL response against the immunodominant H-2D(b) epitope NP366. Finally, the protective potential of the CTL response induced by immunization of mice with rSFV-NP RNA was demonstrated by the reduction of virus load in the lungs after challenge infection with mouse-adapted influenza A/PR/8/34 virus and was comparable to the protective potential of the response induced by plasmid DNA immunization. These results demonstrate that naked RNA immunization with self-replicating molecules can effectively induce both humoral and cellular immune responses and constitutes an alternative strategy to DNA immunization. PMID- 11413387 TI - Characterization of a new H-2D(k)-restricted epitope prominent in primary influenza A virus infection. AB - Influenza A virus infection of mice has been used extensively as a model to investigate the mechanisms of antigen presentation to cytotoxic T lymphocytes (CTL) and the phenomenon of immunodominance in antiviral CTL responses. The different virus-encoded epitopes that are recognized in H-2(b) and H-2(d) mice have been characterized and their relative immunodominance has been well-studied. In H-2(k) mice, four different K(k)-restricted influenza virus epitopes have been described, but the dominance hierarchy of these epitopes is unknown and there is also an uncharacterized D(k)-restricted response against the virus. In this study, a D(k)-restricted epitope derived from the influenza virus A/PR/8/34 polymerase protein PB1, corresponding to amino acid residues 349-357 (ARLGKGYMF), was identified. This peptide is the major epitope within the PB1 polymerase and is at least as dominant as any of the four K(k)-restricted epitopes that are recognized in CBA mice following primary influenza virus infection. The PB1 epitope is only the fourth D(k)-presented peptide to be reported and the sequence of this epitope confirms a D(k)-restricted peptide motif, consisting of arginine at position two, arginine or lysine at position five and a hydrophobic residue at the carboxy terminus. PMID- 11413388 TI - Cloning and identification of the infectious salmon anaemia virus haemagglutinin. AB - Infectious salmon anaemia virus (ISAV) is an orthomyxo-like virus that causes serious disease in Atlantic salmon (Salmo salar). Like the orthomyxoviruses, ISAV has been shown to possess haemagglutinin (HA) activity. This study presents the cloning, expression and identification of the ISAV HA gene, which was isolated from a cDNA library by immunoscreening. The HA gene contained an ISAV-specific conserved nucleotide motif in the 5' region and a 1167 bp open reading frame encoding a protein with a predicted molecular mass of 42.4 kDa. The HA gene was expressed in a baculovirus system. A monoclonal antibody (MAb) shown previously to be directed against the ISAV HA reacted with insect cells infected with recombinant baculovirus. Salmon erythrocytes also adsorbed to these cells and adsorption was inhibited by the addition of either the ISAV-specific MAb or a polyclonal rabbit serum prepared against purified virus, confirming the virus specificity of the reaction. Immunoblot analyses indicated that ISAV HA, in contrast to influenza virus HA, is not posttranslationally cleaved. Sequence comparisons of the HA gene from five Norwegian, one Scottish and one Canadian isolate revealed a highly polymorphic region that may be useful in epidemiological studies. PMID- 11413389 TI - Expression, purification and characterization of the Spodoptera littoralis nucleopolyhedrovirus (SpliNPV) DNA polymerase and interaction with the SpliNPV non-hr origin of DNA replication. AB - The DNA polymerase from Spodoptera littoralis nucleopolyhedrovirus (SpliNPV) was expressed in, and purified from, prokaryotic and eukaryotic expression systems. While less protein was obtained from the E. coli expression system, SpliNPV DNAPOL purified from E. coli displayed similar biochemical characteristics to DNAPOL expressed in, and subsequently purified from, insect cells (Sf9) using a baculovirus expression system. Biochemical analyses suggested that the DNA polymerase and the 3'-5' exonuclease activities are intrinsic to the protein. Deletion of the first 80 amino acid residues from the N terminus of the DNAPOL affected neither the DNA polymerase nor the exonuclease activities of the enzyme. Replication products from single-stranded M13 DNA demonstrated that the DNA synthesis activity of SpliNPV DNAPOL is highly processive. Transient expression assays with a set of deletion clones containing the putative SpliNPV non-hr origin of DNA replication permitted functional characterization of sequence elements within the origin fragment. Purified SpliNPV DNAPOL stimulated origin dependent DNA replication in a cell-free replication assay. PMID- 11413390 TI - Comparative pathogenesis of Helicoverpa zea S nucleopolyhedrovirus in noctuid larvae. AB - We used a recombinant of Helicoverpa zea S nucleopolyhedrovirus containing the hsp70/lacZ reporter cassette (HzSNPV-hsp70/lacZ) to quantify mortality relationships and to elucidate early pathogenesis in two permissive hosts, Heliothis virescens and Helicoverpa zea, and one semi-permissive host, Trichoplusia ni. Fourth instar T. ni were highly resistant to fatal infection both by oral injection of occlusions and by intrahaemocoelic injection of budded virus, indicating the presence of both midgut and systemic mechanisms of resistance. In bioassays, newly moulted (4(0)) H. zea were significantly more susceptible than 4(0) H. virescens to fatal infection, but mortality levels were the same for larval cohorts inoculated 16 h after the moult (4(16)). Developmental resistance was stronger in H. zea and in both hosts, partially reversed by administration of the optical brightener M2R. In both species, developmental resistance was correlated with a reduced ability of HzSNPV to establish and/or maintain primary midgut infections. In time-course experiments using a dosage of 15 occlusions ( approximately LD(90)), lacZ expression marking the onset of primary and secondary infection was first observed in midgut columnar and tracheal cells at 4 and 12 h, respectively. Inoculation of 4(0) larvae resulted in approximately twofold more foci in H. zea larvae than in H. virescens, but H. zea larvae sloughed infected midgut cells at a faster rate. For both heliothines, interaction of occlusion-derived virus with primary cellular targets within the midgut epithelium was critical to the outcome of infection and a key process underlying acquisition of developmental resistance. PMID- 11413391 TI - Genomic organization of RNA2 of Tomato ringspot virus: processing at a third cleavage site in the N-terminal region of the polyprotein in vitro. AB - The proteinase of Tomato ringspot virus (genus Nepovirus) is responsible for proteolytic cleavage of the RNA2-encoded polyprotein (P2) at two cleavage sites, allowing definition of the domains for the movement protein (MP) and coat protein. In this study, we have characterized a third cleavage site in the N terminal region of P2 using an in vitro processing assay and partial cDNA clones. Results from site-directed mutagenesis of putative cleavage sites suggest that cleavage occurs at dipeptide Q(301)/G. Cleavage at this site is predicted to result in the release of two proteins from the N-terminal region of P2: a 34 kDa protein located at the N terminus of P2 (assuming translation initiation at the first AUG codon) and a 71 kDa protein located immediately upstream of the MP domain. In contrast, only one protein domain is present in the equivalent region of the P2 polyprotein of other characterized nepoviruses. PMID- 11413392 TI - Quasispecies nature of the genome of Grapevine fanleaf virus. AB - Genetic diversity was characterized in 14 isolates of Grapevine fanleaf virus (GFLV) recovered from grapevine (Vitis vinifera). Virions were collected by immunocapture, and a 1557 bp fragment containing part of the viral coat protein gene and part of the untranslated region to its 3' side was amplified by RT-PCR. Sequence variation among isolates was characterized by restriction fragment length polymorphism (RFLP) analysis and by sequencing. The AvaII-generated RFLP patterns from the various isolates were highly variable. The isolates were passaged in Chenopodium quinoa. The RFLP patterns altered with passage through the alternate host, but the variation stabilized after a number of passages. Individual genomes were recovered by cloning. The subcloned sequences were found to vary from each other by as much as 13%, and the encoded amino acid sequences by as much as 9%. The data suggest that the GFLV genome consists of quasispecies populations. PMID- 11413393 TI - Peptide display on live MS2 phage: restrictions at the RNA genome level. AB - The potential of the RNA phage MS2 to accommodate extra amino acids in its major coat protein has been examined. Accordingly, a pentapeptide was encoded in the genome as an N-terminal extension. In the MS2 crystal structure, this part of the coat protein forms a loop that extends from the outer surface of the icosahedral virion. At the RNA level, the insert forms a large loop at the top of an existing hairpin. This study shows that it is possible to maintain inserts in the coat protein of live phages. However, not all inserts were genetically stable. Some suffer deletions, while others underwent adaptation by base substitutions. Whether or not an insert is stable appears to be determined by the choice of the nucleic acid sequence used to encode the extra peptide. This effect was not caused by differential translation, because coat-protein synthesis was equal in wild-type and mutants. We conclude that the stability of the insert depends on the structure of the large RNA hairpin loop, as demonstrated by the fact that a single substitution can convert an unstable loop into a stable one. PMID- 11413395 TI - Computerized corneal topography and its importance to wavefront technology. AB - PURPOSE: To review the development and application of corneal topography in refractive surgery. METHODS: Review of the literature and discussion of recent developments in corneal topography and wavefront technology. RESULTS: Analysis of corneal topography provides critical information for the preoperative examination of patients before refractive surgery and for the evaluation and treatment of patients with complications after surgery. CONCLUSIONS: Corneal topography will continue to be a critical diagnostic modality for refractive surgery. Even with the advent of wavefront analysis designed to detect refractive error and aberrations of the eye, it will be necessary to have detailed corneal topographic information to understand the contribution the cornea makes to vision so that custom alteration of that surface can be used to optimize vision. This will be true of the normal eye, but it will be of special importance in eyes with abnormalities that were induced by corneal surgery. PMID- 11413396 TI - Ipsilateral rotational autokeratoplasty: an alternative to penetrating keratoplasty in nonprogressive central corneal scars. AB - PURPOSE: To report the outcome of ipsilateral rotational autokeratoplasty performed for nonprogressive central corneal opacities. METHODS: Medical records of 27 patients who underwent ipsilateral rotational autokeratoplasty between June 1995 and December 1998 were retrospectively reviewed. The etiology of corneal opacity, preoperative visual acuity, and size of the trephine used was noted. Any intraoperative and early and late postoperative complications were noted in all patients. The final outcome, visual acuity, and astigmatism at final correction were analyzed. RESULTS: The main etiology of corneal opacity was trauma (44.4%), followed by resolved keratitis (29.6%). The size of the graft ranged from 6.5 to 9 mm. Fifteen patients underwent autokeratoplasty alone; additional procedures were combined in the remaining 12. Wound leak was the most common postoperative complication, which was seen in seven patients. The mean follow-up was 12.11 months (SD = 11.95 months). Five patients (18.5%) were lost to follow-up. Of the remaining 22, 19 (86.36%) had clear grafts and 3 (13.6%) had failed grafts. The mean astigmatism at final correction was 4.25 diopters cylinder (SD = 2.72). Final visual acuity of 20/80 or better was obtained in 13 cases (59.09%). CONCLUSIONS: Contrary to common belief, ipsilateral rotational autokeratoplasty is a safe and effective surgical procedure. It can be considered as an alternative procedure to penetrating keratoplasty in a scenario in which tissue scarcity exists, as well as in cases that have a high risk of developing immunologic allograft rejection. Further prospective studies with preoperative specular microscopy and postoperative corneal topography will provide greater insight into the usefulness of this procedure. PMID- 11413397 TI - Treatment and outcome of nocardia keratitis. AB - OBJECTIVE: To report our experience in treatment and outcome ofNocardia keratitis. METHODS: Medical and microbiology records of seven cases of culture provenNocardia keratitis seen between January 1997 and March 1999 were reviewed retrospectively. In all patients, corneal scrapings were obtained for direct microscopic evaluation and culture. Drug sensitivity was determined by the Kirby Bauer disk-diffusion method. The minimum inhibitory concentration of ciprofloxacin and amikacin for these isolates was determined by agar dilution method. Response to medical therapy and the end result were analyzed. RESULTS: By the in vitro Kirby-Bauer disk-diffusion techniques, all isolates were sensitive to amikacin; six of these isolates were sensitive to gentamicin and four were sensitive to ciprofloxacin. The minimum inhibitory concentration (MIC) of amikacin for all isolates by the agar-dilution technique was well below the MIC breakpoint forNocardia resistance, whereas the MIC of ciprofloxacin was above the MIC breakpoint forNocardia resistance. All patients responded to medical therapy. The corneal infection resolved in three patients after treatment with ciprofloxacin, in one patient after fortified gentamicin, and in three patients after fortified amikacin. Outcome details were available for six patients. There was good visual recovery in four patients, with visual acuity of 20/25 or better in three. The cornea of two patients developed nonvascularized scars, and in four patients in whom the infiltrates were peripheral, vascularization was seen. CONCLUSIONS: Although patients ofNocardia keratitis may respond to other antibiotics, amikacin appears to be a drug of choice. In this small series, when appropriate therapy was initiated,Nocardia keratitis resolved promptly with good visual recovery. PMID- 11413398 TI - Association of acinetobacter species with contact lens-induced adverse responses. AB - PURPOSE: To determine the levels of Acinetobacter species associated with normal soft contact lens wear and to determine whether Acinetobacter species are involved in adverse reactions that occur during contact lens wear. METHODS: Patients wore soft contact lenses on an extended-wear basis. The bacteria on lenses and ocular swabs during asymptomatic and symptomatic lens wear were identified using standard microbiologic methods. RESULTS: Acinetobacter species were isolated and identified from 16 (13%) of 126 patient samples. Greater numbers of Acinetobacter species were isolated from lenses of patients experiencing adverse responses than from asymptomatic patients. Acinetobacter species were isolated from patients experiencing symptomatic adverse responses in 4 (13%) of 32 cases. CONCLUSION: It appeared that Acinetobacter species colonized the eye of extended contact lens wearers at a time when the normal functioning of the eye was compromised by contact lens wear. When Acinetobacter species were in high numbers on a contact lens, an adverse response occurred. This may implicate Acinetobacter species as a contributing factor to adverse responses associated with contact lens wear. PMID- 11413399 TI - Microwave treatment of contact lens cases contaminated with acanthamoeba. AB - PURPOSE: Microbially contaminated contact lens cases are a predisposing risk factor for Acanthamoeba keratitis. Several findings have shown that microwave irradiation kills the six Food and Drug Administration test challenge microorganisms. We aimed to determine what effect microwave irradiation has on Acanthamoeba trophozoites and cysts. METHODS: Different types of contact lens cases were contaminated with trophozoites and cysts of three different Acanthamoeba species (A. comandoni, A. castellanii, A. hatchetti) and were exposed to microwave irradiation for 3, 5, and 8 minutes, respectively. RESULTS: Trophozoites, as well as cysts of the different Acanthamoeba strains, were effectively killed, even by only 3 minutes of microwave irradiation, and there were no negative effects of irradiation on the contact lens cases themselves. CONCLUSION: We demonstrate that microwave treatment is a very effective, easy, and cheap method to keep contact lens cases free of Acanthamoeba, thus considerably reducing the risk of an Acanthamoeba keratitis. PMID- 11413400 TI - Binocular vision recovery in bilateral keratoplasty. AB - PURPOSE: To determine the benefit of bilateral penetrating keratoplasty with regard to binocular vision. METHODS: We compared patients who underwent corneal transplantation bilaterally with patients who had successful corneal transplantation in one eye and corneal disease in the other eye. Specifically, changes in fusion, stereopsis, and binocular vision function were analyzed. A questionnaire regarding performance of daily tasks was given. RESULTS: Patients with bilateral keratoplasty performed better in all the analyzed functions. Fusion was achieved by 81.25% in the unilateral group versus 100% in the bilateral group (p = 0.15). Stereopsis was present more in the bilateral group (100% vs. 62.5%, p = 0.008) and the quantity of stereopsis was significantly better in the bilateral group (121 seconds of arc vs. 1,284 seconds of arc, p = 0.014). 88.8% of the patients subjectively improved in daily activities after second eye surgery. CONCLUSION: There are objective and subjective improvements after bilateral penetrating keratoplasty. PMID- 11413401 TI - The effect of state legislation on eye donation. AB - PURPOSE: Pennsylvania Act 102 implemented in March 1995 required all acute care hospitals in Pennsylvania to routinely refer all deaths to the Organ Procurement Organization for determination of suitability for organ/tissue donation. This study analyzed the effect of the law on eye donation. METHODS: Retrospective analysis of the total number of referrals and the actual number of eye donations from 62 hospitals in Pennsylvania to the Lions Eye Bank of Delaware Valley was performed for the years 1993 to 1998. Information gathered included donor's age, gender, race, cause of death, referring institution, and result of referral. RESULTS: From 1993 to 1998, the total numbers of referrals were 988, 1,647, 8,101, 21,123, 21,783, and 22,987, and the numbers of donors were 570, 574, 660, 644, 594, and 568, respectively. The increase in the number of donors after implementation of the law was not commensurate with the number of referrals. This was caused by a disproportionate increase in the number of referrals older than 70 years of age (from a mean of 33% to 52%), which exceeded the donor age limit of 69 years, and also to a lower family consent rate (from a mean of 48% to 24%). CONCLUSIONS: Well-designed state legislation with proper implementation greatly increased hospital referrals for eye donation. However, there was only a small increase in the number of eye donors because many of the referrals were beyond the acceptable upper age limit for eye donation. A small increase in the donor age limit would increase the number of eye donations without having to expand the potential donor pool. Education of the public may help to improve the family consent rate. PMID- 11413402 TI - Diurnal variation of corneal and corneal epithelial thickness measured using optical coherence tomography. AB - PURPOSE: To determine the feasibility of optical coherence tomography (OCT) measurement of overnight corneal swelling and de-swelling and, specifically, to examine overnight changes in the corneal epithelium and recovery during the day. METHODS: Ten healthy volunteers were recruited. Corneal OCT imaging was performed at 10:00 PM (baseline), 8:30 AM, and every 2 hours thereafter until 4:30 PM. Either the right or left eye was taped shut (using a gauze pad and surgical tape) until the 8:30 AM session of the next day. Measurements were made along a 1-mm strip centered on the visual axis. RESULTS: At baseline, there were no differences in corneal or epithelial thickness between control and experimental eyes (all p > 0.05). Immediately after patch removal, corneal and epithelial thicknesses of patched eyes were higher than the baseline (all p < 0.05). In patched eyes, the cornea and epithelium swelled 5.5% and 8.1% overnight, respectively (p > 0.05). Two hours later, the epithelial thickness of the experimental eyes recovered to baseline level (p > 0.05), but corneal thickness did not reach to baseline level until 4 hours after patch removal. For control eyes, there were no differences compared with baseline (all p > 0.05). CONCLUSION: Both cornea and corneal epithelium experience proportionately similar amounts of overnight swelling. Recovery of overnight swelling may be slower for the cornea than for the epithelium. OCT provides valuable information about anterior segment morphometry. PMID- 11413403 TI - Spreading of the tears after a blink: velocity and stabilization time in healthy eyes. AB - PURPOSE: To quantify the tear spreading velocity and stabilization time after blinks in healthy patients. METHODS: Video recordings (magnification, x130) were made of the movements of naturally occurring particles in the tear film while patients blinked naturally. After each blink, tear spreading velocity was computed every 40 milliseconds from particle displacements determined from a frame-by-frame analysis of the records. RESULTS: After a blink, tears moved rapidly upward over the cornea. Forty milliseconds after passage of the lid, the velocity was 7.34 +/- 2.73 mm/s (mean +/- standard deviation, n = 20). The time to tear stabilization (i.e., zero velocity) was 1.05 +/- 0.30 seconds. The decay of spreading velocity with time was well described by a logarithmic function for all individual patient data (R2 range, 0.88-0.99; n = 20). We have shown that initial velocity and stabilization time are independent descriptors of tear spreading. Meibomian gland expression markedly reduced initial velocity leaving a normal stabilization time, whereas inhalation of an irritant reduced stabilization time leaving a normal initial velocity. In a patient with Sjogren syndrome, punctal plugs rapidly restored initial velocity and stabilization time from near zero values to normal values. CONCLUSION: We provide normal values for two descriptors of tear spreading, namely initial velocity and time to stabilization. These values can be modified by manipulations that alter tear constituents. Consequently, these descriptors may provide a basis for quantitative, noninvasive tear assessment. PMID- 11413404 TI - The potential for eye bank limbal rings to generate cultured corneal epithelial allografts. AB - PURPOSE: Patients with severe limbal deficiencies are unable to maintain a stable corneal surface. If sheets of cultured allogeneic corneal epithelium could be prepared from eye banked corneal limbal rings, which are normally discarded after keratoplasty, the sheets may be beneficial for grafting onto patients with limbal stem cell deficiencies. METHOD: Biopsies of limbal tissue (2-3 mm2) removed from organ-cultured corneal limbal rings or from fresh whole globes were either trypsinized or set up as explants to assess their potential for corneal epithelial cell production. RESULTS: Several biopsies were taken from each of 21 organ-cultured limbal rings and 10 fresh cadaveric globes. Cultures were generated from every cadaveric eye (10/10), although not all biopsies from the same eye gave rise to cultures. Confluent sheets of cultured cells were also produced successfully from limbal rings that had been in organ culture for up to 25 days, but the success rate from limbal ring material was variable (14/21). An analysis of parameters associated with each limbal ring was carried out in an attempt to identify the reasons for the different efficiencies of epithelial production. No obvious single parameter correlation was detected, although there was a trend to poorer efficiency with increased donor age. CONCLUSIONS: Confluent sheets of cultured corneal epithelial cells, suitable for grafting, can be produced from limbal tissue taken from eye bank organ-cultured corneas, although it takes longer, on average, to reach confluence (17-21 days) than an equivalent sample from a fresh eye (9-12 days). PMID- 11413405 TI - Efficacy of polyclonal antibodies for treatment of ocular herpes simplex infection. AB - PURPOSE: Herpes simplex virus (HSV) can cause corneal infections in humans and lead to permanent scarring, loss of vision, and blindness. Current treatment of epithelial HSV keratitis consists of using antiviral DNA analogs. In this study, we used in vitro and in vivo models to evaluate the efficacy of six polyclonal antibodies to HSV recombinant surface glycoprotein D in treating ocular epithelial HSV. METHODS: Confluent cultures of African Green monkey kidney fibroblasts (Vero cells) and normal 3-to 5-lb female New Zealand White rabbits were infected with HSV type 1, strain RE. In vitro virucidal and antiviral assays were performed, and the best of the compounds was chosen for the in vivo stage. Animals were carefully monitored until day 5 after HSV-1 inoculation, then arbitrarily divided into groups receiving, for 14 days, varying doses of: polyclonal antibodies four times a day, polyclonal antibodies three times a day, trifluorothymidine (current treatment of choice and the positive control) nine times a day, or 0.9% physiologic saline nine times a day. The animals were followed up in a masked fashion and carefully monitored for severity and resolution of the HSV infection by biomicroscopy (slit lamp) examination and viral cultures using standardized plaque assays. RESULTS: All six of the compounds tested were effective in vitro, but one compound in particular, SP-510 50, was superior. It was used for the in vivo testing and showed antiviral efficacy in a dose-dependent manner, and at dosing four times a day, it was of comparable efficacy to trifluorothymidine (nine times a day). CONCLUSIONS: We conclude that polyclonal antibodies to glycoprotein D appear to be effective antiviral agents in vitro and in vivo in a rabbit model of HSV-1 keratitis and show promise as a new antiviral treatment for ophthalmic use. PMID- 11413406 TI - Corneal temperature reversal after storage in Chen medium compared with Optisol GS. AB - PURPOSE: To compare corneal endothelial cell function by measuring corneal thickness during temperature reversal between corneas stored in two different storage media, Optisol GS and Chen Medium (CM). METHODS: Twenty paired corneas from 10 human donors were randomly assigned for storage at 4 degrees C in Optisol GS (10 corneas) or CM (10 corneas). The storage media were masked, and measurements were done in a masked fashion. After storage for 48 hours, corneal thickness was measured by ultrasonic pachymetry at 2-hour intervals for 12 hours, during which time the corneas were perfused with BSS (balanced salt solution) Plus at 37 degrees C. Scanning electron microscopy of two pairs of corneas from two donors was performed to assess ultrastructural change after 12 hours of warming. RESULTS: Corneal thickness decreased during the first 4 hours of the warming period and then increased during the 6-to 12-hour warming period. These changes in corneal thickness over time were similar for the two storage media (p = 0.212). Scanning electron microscopy showed greater amounts of endothelial cell disruption in Optisol GS-stored corneas than those stored in CM after 12 hours of warming and perfusion. CONCLUSIONS: The endothelial pump of corneas stored in CM appear to be as well-preserved as those stored in Optisol GS, although greater endothelial disruption may be present with Optisol GS by scanning electron microscopy. Further studies are required to compare the clinical effectiveness of these two media. PMID- 11413407 TI - Effects of three penetration enhancers on transcorneal permeation of cyclosporine. AB - PURPOSE: To investigate the permeation of cyclosporin A (CsA) through fresh and frozen human corneas in the presence and absence of three penetration enhancers: benzalkonium chloride (BZCl), dimethylsulfoxide (DMSO), and Cremophor-EL. METHODS: Human corneas, unsuitable for transplantation, were either freshly used for permeability experiments or snap-frozen in liquid nitrogen and stored at -85 degrees C. CsA permeation through either fresh or thawed frozen corneal tissue was determined using a flow-through diffusion apparatus (20 degrees C for 24 hours). Flux rates for CsA were determined in the presence and absence of the penetration enhancers 0.01% BZCl, 20% DMSO, and Cremophor-EL (10% and 20%). Analysis of variance and Duncan's multiple-range test were used to test for steady state, and an unpaired Student t test with Welch's correction was used to test for differences between the mean flux values at each time point. A significance level of 5% was used for all of the statistical tests. RESULTS: No statistically significant differences in flux values of CsA could be detected between fresh and frozen corneas. In the presence of Cremophor-EL (10% and 20%) and 0.01% BZCl, statistically significant increases in flux values of CsA before 16 hours and after 16 hours, respectively, could be observed. In the presence of 20% DMSO, no statistically significant increases in flux values could be detected. CONCLUSIONS: The permeation of CsA through human corneas appeared to be enhanced by the presence of BZCl and Cremophor-EL. Additionally, it was shown that the flux rate of CsA across fresh and frozen corneas was not significantly different. PMID- 11413408 TI - Thermal consequences of photorefractive keratectomy. AB - PURPOSE: Photorefractive keratectomy (PRK) is still a widely used method for the correction of refractive error despite the advent of laser in situ keratomileusis (LASIK). However, both procedures are associated with significant side effects such as corneal haze and regression. Several factors have been implicated in the etiology of haze, one of which is thermal loading of the cornea. The purpose of this study was to investigate the temperature changes occurring during PRK when performed at different ablation depths. METHODS: Noncontact, color-coded ocular thermography was performed with an infrared detector apparatus during PRK ablation on 19 ovine corneas. Five different refractive corrections were carried out ranging from -2.00 diopters (D) (ablation depth, 31.2 microm) to -10.00 D (ablation depth, 137.9 microm). RESULTS: A temperature rise at the corneal surface was demonstrated in all 19 corneas. The mean rise in temperature was 7.35 +/- 1.13 degrees C with a maximum rise in temperature of 8.97 degrees C. A positive correlation was found between the refractive correction and the peak rise in temperature (r2 = 0.57, p< 0.0001). The rate of temperature change was greater for smaller treatments than for larger treatments (r2 = 0.79, p < 0.0001). Corneas undergoing larger treatments were subject to greater rises in temperature for longer periods of time. CONCLUSIONS: This study suggests that the cornea undergoes a significant rise in temperature as a result of the PRK process. Further investigation is required to determine what effect this thermal loading has on the corneal wound healing response after PRK. PMID- 11413409 TI - The eicosanoid, 15-(S)-HETE, stimulates secretion of mucin-like glycoprotein by the corneal epithelium. AB - PURPOSE: The eicosanoid, 15-(S)-hydroxyeicosa-5Z, 8Z-11Z, 13E-tetraenoic acid (15 (S)-HETE), is known to stimulate production of mucin glycoprotein by airway epithelium. This study investigated the effect of 15-(S)-HETE on the mucin glycoprotein secretion by the corneal epithelium. METHODS: To determine the effect of dose, corneas of anesthetized New Zealand White rabbits were treated with 50, 500, or 5,000 nM 15-(S)-HETE in artificial tears for 120 minutes. To determine the time to onset of the response, corneas were treated with 500 or 1,000 nM 15-(S)-HETE in balanced salt solution for periods ranging from 5 to 120 minutes. Corneas were fixed for electron microscopy in fixative containing 0.5% cetylpyridinium chloride (CPC) to stabilize the layer of mucin-like glycoprotein on the corneal surface. The mucin layer thickness was measured by image analysis of electron micrographs. RESULTS: The layer of CPC-fixed mucin-like glycoprotein on the surface of control corneas was 0.46 +/- 0.04 microm thick. After treatment with 15-(S)-HETE, the thickness of the mucin layer increased to 0.64 +/- 0.1 microm at 50 or 5,000 nM HETE and as much as 1.02 +/- 0.2 microm in response to 500 nM HETE. Mucin thickness reached a statistical maximum of 0.59 +/- 0.1 microm after only 5 minutes of exposure to 500 or 1,000 nM HETE. CONCLUSIONS: Exposure of the cornea to 15-(S)-HETE causes a rapid-onset increase in the thickness of a layer of mucin-like glycoprotein on the surface of the corneal epithelium. This supports previous reports that corneal epithelial cells produce mucin and suggests that treatment with topical 15-(S)-HETE may be effective in treating ocular surface mucin deficiency in dry eye syndrome. PMID- 11413410 TI - Peripheral nonulcerative keratitis as a presenting sign of microscopic polyangiitis. AB - PURPOSE: To report a 16-year-old woman who had peripheral keratitis as a presenting sign of microscopic polyangiitis (MPA), which rapidly progressed to acute renal failure. METHOD: Case report. RESULTS: The patient's vasculitis was diagnosed by renal biopsy, which was evaluated with histologic, immunofluorescence, and electron microscopy. The diagnosis was confirmed by laboratory study, which showed a positive antimyeloperoxidase antibody. CONCLUSION: MPA may rarely present with ocular findings and should be considered in the differential diagnosis when a patient has peripheral keratitis. PMID- 11413411 TI - Late-onset form of lattice corneal dystrophy caused by leu527Arg mutation of the TGFBI gene. AB - PURPOSE: To report two Japanese patients who were clinically diagnosed with late onset and sporadic lattice corneal dystrophy (LCD) in whom a Leu527Arg mutation in the TGFBI gene was found. METHODS: Molecular genetic analysis was performed on DNA extracted from peripheral leukocytes from the patients. Exons 4, 11, and 12 of the TGFBI gene were amplified by polymerase chain reaction and directly sequenced. Histopathologic study was performed on the corneal tissue obtained during deep lamellar keratoplasty (DLK) from one of the patients. RESULTS: Patient 1 was a 74-year-old man who noticed a visual disturbance at the age of 72 years. Deep stromal opacities with nodular deposits and thick lattice lines were observed only in the right cornea, and DLK was performed. Patient 2 was an 82 year-old man who had LCD (similar in appearance to that in patient 1) in both eyes without visual disturbance. Neither of the patients had a family history of corneal problems and had no episode of corneal erosion. A heterozygous single base-pair transition (CTG to CGG, leucine to arginin) was detected in codon 527 of the TGFBI gene in both patients. No mutation was found in codons 124, 501, 518, 546, or 555. Histopathologically, relatively large amyloid deposits in the deep corneal stroma and ribbons of amyloid deposits just beneath the Bowman's layer were observed in the corneal tissue of patient 1. CONCLUSIONS: Clinical features and pathologic findings of the late-onset form of LCD with an L527R mutation in the TGFBI gene were made clear. PMID- 11413412 TI - Triple anterior chamber after full-thickness lamellar keratoplasty for lattice corneal dystrophy. AB - PURPOSE: To report a patient with lattice corneal dystrophy type I (LCDI) who developed a triple anterior chamber after full-thickness lamellar keratoplasty (LKP). METHODS: A 46-year-old woman underwent a full-thickness LKP in her right eye for visual disturbances caused by LCDI. Her visual acuity was 20/200 OD before surgery. A complete ophthalmic examination, including slit lamp biomicroscopy and optical coherence tomography (OCT), was performed before and after surgery. Molecular genetic analysis was performed on DNA extracted from the peripheral leukocytes. RESULTS: The surgery was performed uneventfully; however, extra spaces posterior to the graft, along with the severe graft edema, were observed to form a triple anterior chamber a few days after surgery. The extra spaces resolved in 3 weeks with no surgical treatment, and her visual acuity improved to 20/20 OD without correction 3 months after surgery. The triple anterior chamber was clearly demonstrated by OCT, but not by slit lamp biomicroscopy. A heterozygous single base-pair transition (CGC to TGC, arginin to cysteine) was detected in codon 124 of the TGFBIgene in the patient. CONCLUSION: The separation of the graft and the host's deep corneal tissue and a Descemet's membrane detachment in the host's cornea caused the triple anterior chamber. The Descemet's membrane detachment demonstrated the weak adhesion of the stroma and the Descemet's membrane, probably resulting from a dysfunction of the TGFBI protein caused by the mutation of the TGFBIgene. OCT is useful for the objective documentation of the posterior corneal region even with severe corneal edema. PMID- 11413413 TI - Aspergillus fumigatus keratitis with wreath pattern infiltrates. AB - PURPOSE: To report a case of Aspergillus fumigatus keratitis with clinical features simulating Nocardia keratitis and to highlight the utility of microbiologic investigation in the successful management of infectious keratitis. METHOD: Case report. RESULTS: A 62-year-old man presented with complaints of pain, redness, and watering of 10 days' duration in his right eye. Direct microscopic observation of smears of corneal scrapings revealed a fungal etiology. The patient was treated with 5% natamycin eye drops and 1% atropine sulphate eye drops and was advised to visit the hospital for observation. During his visit to the hospital on day 10 after medication, the eye demonstrated a wreath pattern corneal infiltrate that simulated Nocardia keratitis. The fungus grown from culture of corneal scraping was identified as A. fumigatus. CONCLUSION: This report highlights the significance of subjecting corneal scrapings from suspected cases of infectious keratitis to microbiologic evaluation and emphasizes the fact that a complete microbiologic work-up helps in establishing a definitive etiologic diagnosis and initiating specific antimicrobial therapy. PMID- 11413414 TI - Mucolipidosis IV in an African American patient with new findings on electron microscopy. AB - PURPOSE: We report an unusual case of mucolipidosis IV in a patient of African ancestry, with intracytoplasmic inclusions of the corneal endothelium found on electron microscopy. METHOD: Clinical description with light and electron microscopy. RESULTS: We describe a case of mucolipidosis IV diagnosed in a patient of African ancestry after penetrating keratoplasty. Electron microscopic evaluation revealed intracytoplasmic inclusions in both the corneal epithelium and endothelium. CONCLUSION: The diagnosis of mucolipidosis in a patient of African ancestry is unusual, as this genetic disorder is found predominantly in individuals of Jewish descent. Corneal endothelial involvement in mucolipidosis IV has not previously been reported. PMID- 11413415 TI - A case of atypical Cogan's syndrome with uncommon corneal findings. AB - PURPOSE: We report a case of atypical bilateral interstitial keratitis associated with Cogan's syndrome. METHODS: A 28-year-old man presented with a 2-year history of recurrent bilateral keratitis. Bilateral hearing loss preceded the ocular symptoms by 2 years. The patient also complained of skin nodules, headache, back pain, and arthritis. Corneal finding were consistent with superior stromal keratitis with stromal neovascularization and lipid deposition in the stroma. The patient's audiogram revealed cochlear pathology compatible with Cogan's syndrome (sensorineural deafness). RESULTS: The patient was treated with topical steroids but eventually required corneal transplantation in the right eye as a consequence of progressive loss of vision secondary to progressive lipid keratopathy. Visual acuity at the patient's most recent follow-up evaluation was 20/40. CONCLUSION: This case represents an unusual type of interstitial keratitis associated with Cogan's disease. The absence of ocular symptoms at the time of initial ear involvement and the atypical presentation of the keratitis were responsible for the delay in diagnosis in this patient, resulting in hearing impairment. PMID- 11413416 TI - Conjunctival necrobiotic granuloma in xeroderma pigmentosum. AB - PURPOSE: We report clinical and histopathologic findings of a conjunctival lesion associated with xeroderma pigmentosum. METHODS: A Saudi girl with known xeroderma pigmentosum presented with pain and photophobia of the right eye and an elevated temporally located perilimbal mass. RESULTS: The mass was resected successfully and has not recurred during 1-year follow-up. Histopathologic examination of the tissue showed a necrobiotic granuloma with associated histiocytic infiltration. The patient had no systemic disease, lipid was not detected in the histiocytic lesion, no Touton giant cells were present, and there was no evidence of elastolysis. CONCLUSION: Ocular malignancies occur in association with xeroderma pigmentosum, but benign lesions that mimic a malignancy may occur. PMID- 11413417 TI - Congenital hereditary endothelial dystrophy and band keratopathy in an infant with corpus callosum agenesis. AB - PURPOSE: To report the features of a syndrome of endothelial failure and band shaped keratopathy in an infant with corpus callosum agenesis. METHODS: The clinical and histopathologic features of an infant presenting with bilateral corneal clouding and corpus callosum agenesis are reported. The patient underwent bilateral penetrating keratoplasty at ages 28 months and 4 years. Light and electron microscopy were used to characterize the structural changes. RESULTS: The epithelium was thin and degenerate. Bowman's membrane contained spherical aggregates that were present also within a connective tissue pannus. The midstroma was normal, but there were floral and rope-like aggregations of collagen in the pre-Descemet's membrane region. Under electron microscopy, the spherules formed target-shaped lesions with a central focus of alternating electron-dense and lucent material. Numerous microfilaments in the pannus and anterior stroma labeled with fibrillin-1 antibody. Microfilaments within fibroblasts were stained with vimentin antibody. Posteriorly, the endothelium was mainly absent and Descemet's membrane showed a fetal layer and a posterior collagenous layer. CONCLUSION: Corneal appearances in this patient were in keeping with those of congenital hereditary endothelial corneal dystrophy. However, there was no family history and neither parent showed a clinical endothelial abnormality. The presence of fetal, banded material in Descemet's membrane suggested that endothelial loss began at or near the time of birth. The band keratopathy was regarded as a secondary change. The association with corpus callosum agenesis does not appear to have been described previously. PMID- 11413418 TI - Multifocal corneal argyrosis after an explosion injury. AB - PURPOSE: To document the clinical and histopathologic corneal features of a patient who developed multifocal corneal argyrosis after a chemical explosion injury with unusual involvement of the corneal stroma and keratocytes. METHODS: The corneal button was investigated by light and transmission electron microscopy and scanning electron microscopy combined with energy-dispersive x-ray microanalysis. RESULTS: Clinically, the patient showed dark discoloration of the lids, periocular skin, episclera, and conjunctiva and had multiple brown dots in the superficial layers of the cornea. Microscopic examination of the cornea showed diffuse deposition of silver particles in the epithelial basement membrane, Bowman's layer, and Descemet's membrane. In the corneal stroma, silver granules accumulated intracellularly within lysosomal structures of degenerative keratocytes and extracellularly in association with collagen fibers and cellular debris. Energy-dispersive x-ray analysis showed peaks of silver and sulfur. CONCLUSION: The toxic influence of intracellular accumulation of silver in stromal keratocytes may lead to cell damage and necrosis and result in visual impairment. PMID- 11413420 TI - Primary sensory neurons with dichotomizing axons projecting to the facet joint and the sciatic nerve in rats. AB - STUDY DESIGN: Dorsal root ganglion (DRG) neurons that have dichotomizing axons to the lumbar facet joint and to the sciatic nerve were investigated in rats using a double fluorescent labeling technique. OBJECTIVES: To clarify the existence of DRG neurons with dichotomizing axons projecting to the lumbar facet joint and to the sciatic nerve in rats. SUMMARY OF BACKGROUND DATA: DRG neurons having dichotomizing axons have been reported in several species and are considered to be related to referred pain. However, such DRG neurons have not been investigated in the lumbar spine. Clinically, pain from the lumbar facet joint is sometimes referred to the lower extremities innervated by the sciatic nerve. METHODS: Two kinds of neurotracers (DiI and FG) were used in the present double-labeling study. DiI crystals were placed in the left L5-L6 facet joint, and FG was applied to the ipsilateral sciatic nerve or along the midline of the L5 dermatome. Bilateral DRGs T13-S1 were observed by fluorescence microscope. RESULTS: DRG neurons double labeled with DiI and FG were recognized only in the ipsilateral DRGs from L3 to L6 levels. Approximately 3% of DRG neurons innervating the L5-L6 facet joint had other axons to the sciatic nerve. By contrast, no double-labeled neurons were observed after FG was applied to the L5 dermatome. CONCLUSIONS: In rats approximately 3% of DRG neurons innervating the lumbar facet joints have dichotomized axons projecting to the sciatic nerve. PMID- 11413421 TI - Herniation of cervical intervertebral disc: immunohistochemical examination and measurement of nitric oxide production. AB - STUDY DESIGN: Surgically obtained cervical herniated intervertebral discs were examined histologically and immunohistochemically. The production of nitric oxide (NO) in the local tissue was examined using the electron spin resonance (ESR) method. OBJECTIVES: To investigate the local histologic and immunohistochemical changes in cervical disc herniation, including NO production, and to compare such changes with those in autopsy cases. SUMMARY OF BACKGROUND DATA: Very little is known about the histopathologic processes of cervical disc herniation. In addition, no information is available on the level of in vivo NO production in cervical disc herniation. METHODS: Thirty-six herniated cervical discs obtained from 31 patients were immunohistochemically examined for localization of blood vessels, matrix metalloproteinase (MMP)-3, and inducible NO synthetase (iNOS). We also compared the production of NO, measured by the ESR method, in eight specimens with that of five control discs obtained from fresh cadavers. RESULTS: The presence of herniated discs correlated with the degeneration of cartilaginous endplate and torn anulus fibrosus. Formation of new blood vessels around the herniated discs was detected, using von Willebrand factor antibody, in seven uncontained hernias and 20 contained hernias. Immunohistochemical studies showed the presence of cells positive for MMP-3 (chondrocytes), iNOS (chondrocytes and granulation tissue) in cervical disc hernias. ESR analysis showed a significantly higher NO production in herniated cervical discs than in disc samples of fresh cadavers. CONCLUSIONS: Herniated cervical intervertebral disc is characterized by the presence of an inflammatory process associated with neovascularization and increased expression of MMP-3. Production of NO was markedly high in both contained- and uncontained-type hernias. PMID- 11413422 TI - Electromyographic reflex responses to mechanical force, manually assisted spinal manipulative therapy. AB - STUDY DESIGN: Surface electromyographic reflex responses associated with mechanical force, manually assisted (MFMA) spinal manipulative therapy were analyzed in this prospective clinical investigation of 20 consecutive patients with low back pain. OBJECTIVES: To characterize and determine the magnitude of electromyographic reflex responses in human paraspinal muscles during high loading rate mechanical force, manually assisted spinal manipulative therapy of the thoracolumbar spine and sacroiliac joints. SUMMARY OF BACKGROUND DATA: Spinal manipulative therapy has been investigated for its effectiveness in the treatment of patients with low back pain, but its physiologic mechanisms are not well understood. Noteworthy is the fact that spinal manipulative therapy has been demonstrated to produce consistent reflex responses in the back musculature; however, no study has examined the extent of reflex responses in patients with low back pain. METHODS: Twenty patients (10 male and 10 female, mean age 43.0 years) underwent standard physical examination on presentation to an outpatient chiropractic clinic. After repeated isometric trunk extension strength tests, short duration (<5 msec), localized posteroanterior manipulative thrusts were delivered to the sacroiliac joints, and L5, L4, L2, T12, and T8 spinous processes and transverse processes. Surface, linear-enveloped electromyographic (sEMG) recordings were obtained from electrodes located bilaterally over the L5 and L3 erector spinae musculature. Force-time and sEMG time histories were recorded simultaneously to quantify the association between spinal manipulative therapy mechanical and electromyographic response. A total of 1600 sEMG recordings were analyzed from 20 spinal manipulative therapy treatments, and comparisons were made between segmental level, segmental contact point (spinous vs. transverse processes), and magnitude of the reflex response (peak-peak [p-p] ratio and relative mean sEMG). Positive sEMG responses were defined as >2.5 p-p baseline sEMG output (>3.5% relative mean sEMG output). SEMG threshold was further assessed for correlation of patient self-reported pain and disability. RESULTS: Consistent, but relatively localized, reflex responses occurred in response to the localized, brief duration MFMA thrusts delivered to the thoracolumbar spine and SI joints. The time to peak tension (sEMG magnitude) ranged from 50 to 200 msec, and the reflex response times ranged from 2 to 4 msec, the latter consistent with intraspinal conduction times. Overall, the 20 treatments produced systematic and significantly different L5 and L3 sEMG responses, particularly for thrusts delivered to the lumbosacral spine. Thrusts applied over the transverse processes produced more positive sEMG responses (25.4%) in comparison with thrusts applied over the spinous processes (20.6%). Left side thrusts and right side thrusts over the transverse processes elicited positive contralateral L5 and L3 sEMG responses. When the data were examined across both treatment level and electrode site (L5 or L3, L or R), 95% of patients showed positive sEMG response to MFMA thrusts. Patients with frequent to constant low back pain symptoms tended to have a more marked sEMG response in comparison with patients with occasional to intermittent low back pain. CONCLUSIONS: This is the first study demonstrating neuromuscular reflex responses associated with MFMA spinal manipulative therapy in patients with low back pain. Noteworthy was the finding that such mechanical stimulation of both the paraspinal musculature (transverse processes) and spinous processes produced consistent, generally localized sEMG responses. Identification of neuromuscular characteristics, together with a comprehensive assessment of patient clinical status, may provide for clarification of the significance of spinal manipulative therapy in eliciting putative conservative therapeutic benefits in patients with pain of musculoskeletal origin. PMID- 11413423 TI - Flexibility analysis of posterolateral fusions in a New Zealand white rabbit model. AB - STUDY DESIGN: Biomechanics of posterolateral spinal fusion were studied in an in vivo rabbit model. OBJECTIVES: To determine the extent of stabilization produced by posterolateral lumbar fusion and to test the hypothesis that motions are not completely eliminated after successful fusion. SUMMARY OF BACKGROUND DATA: Previous human cadaveric studies, clinical studies, and animal studies have attempted to characterize the biomechanics of posterolateral fusion. Such studies have been limited by either methods of fusion modeling or methods of stability testing. No previous study has examined biologic fusion with a physiologic biomechanical testing technique. METHODS: Ten adult New Zealand white rabbits underwent L5-L6 intertransverse process fusion using autogenous iliac crest bone graft. Rabbits were killed 5 weeks after surgery. Only one time point was studied. This time point was chosen because previous pull-apart studies have shown plateauing of rabbit fusion mass strength and stiffness around this time. Spines were then harvested and evaluated with manual palpation and an established flexibility testing protocol. Resulting data were compared with previously acquired, nonoperative spine flexibility data. RESULTS: Two animals were excluded because of complications. Of those that were fused (n = 5), biomechanical testing revealed significant decreases in flexion (81%), extension (61%), and right and left lateral bending (67% and 83%, respectively) (P < 0.01). CONCLUSIONS: These findings define the amount of motion reduction that can be expected with posterolateral fusions in the rabbit model at 5 weeks. These results suggest that motion was significantly decreased but was not eliminated. PMID- 11413424 TI - Return to play after cervical spine injury. AB - STUDY DESIGN: A questionnaire survey was mailed to members of the Cervical Spine Research Society, the Herodiuus Sports Medicine Society, and to members of the authors' Department of Orthopaedics. OBJECTIVES: The purpose of our study was to evaluate what influence, if any, factors such as published guidelines, type of sport of the patient, number of years in practice, subspecialty interest, and sports participation of the respondent held in the "return to play" decision making process after a cervical spine injury. SUMMARY OF BACKGROUND DATA: The consequences of cervical spine injury are potentially catastrophic, and return to play decisions in athletes with a history of neck injury can be agonizing. Although recent publications have addressed some of the concerns regarding cervical spine injuries in the athletic population, many questions remain unanswered. Factors such as published guidelines, type of sport of the patient, number of years in practice, subspecialty interest, and sports participation of the respondent have all been suggested as having a possible role in return to play decisions. METHODS: Representative radiographs and case histories of 10 athletes who had sustained neck injury were mailed to 346 physicians. For each case physicians selected every type of play (of six categories) that they felt comfortable recommending. Type of play was divided into six categories: Type 1, collision sports; Type 2, contact sports; Type 3, noncontact, high velocity sports; Type 4, noncontact, repetitive load sports (e.g., running); Type 5, noncontact, low impact sports; Type 6, no sports. In addition, demographic data regarding board certification, subspecialty interest, number of years in practice, use of guidelines in return to play decisions, and personal participation in sports were queried from all respondents. Statistical analysis was completed with Statview (Berkeley, CA). Basic descriptive statistics, chi2, and ANOVA were used where appropriate. RESULTS: Three hundred forty-six questionnaires were mailed and 113 were returned (response rate 32.7%). One hundred ten (97%) of the respondents who completed the questionnaire were board certified. Seventy-five were subspecialists in spine, 22 were subspecialists in sportsmedicine, and 13 reported interests in both sports medicine and spine. Use of Published Guidelines. Although 49% of respondents reported using guidelines in decision-making, the use of guidelines was statistically significant in only one case (P = 0.04). Hierarchy of Risk. In general, those physicians who participated in the study followed the hierarchy of risk that we established in this study (Type 1 [collision sports; highest level of risk] through Type 6 [no sports; lowest level of risk]). Twelve (10.6%) respondents, however, deviated from it in one or more cases. Years in Practice. In three cases there was a statistically significant association between the number of years a physician was in practice and the type of play selected (P < 0.05). In each case a lower level of play tended to be recommended by more senior physicians. Subspecialty Interest. In three cases those respondents with a spine subspecialty interest recommended returned to a higher level of play (P < 0.05). CONCLUSIONS: There is no consensus on the postinjury management of many cervical spine-injured patients. Further research, education, and discussion on this topic are needed. PMID- 11413426 TI - Lumbar sagittal alignment after fusion with a threaded interbody cage. AB - STUDY DESIGN: Records of 111 patients randomly selected from a population who received an interbody fusion cage during a clinical Investigation Device Exemption trial (BAK/L) yielded 126 operative levels and were retrospectively assessed. OBJECTIVES: This study examined lumbar spine sagittal alignment and clinical outcomes before and 2 years after fusion surgery. SUMMARY OF BACKGROUND DATA: Lumbar lordosis is important in spinal sagittal alignment and balance, especially the L4-S1 area. Perceived consensus is that anatomically correct lumbar lordosis is desired and that a loss of lumbar lordosis may lead to spine pathology. There is little information on lumbar lordosis after interbody fusion. METHODS: A random sample of 111 patients who received a cylindrical cage implant (total pool of 947 patients) yielded 126 operative lumbar segments. There were 52 posterior approaches and 59 anterior approaches, and all cages were placed in the L4-L5 or L5-S1 levels. Preoperative and 2-year follow-up lateral radiographs were measured for segmental lordosis. Cage position was measured relative to the posterior longitudinal ligament. Segmental lordotic change was correlated to clinical outcome at the 2-year follow-up. RESULTS: Preoperative lordosis was different as a function of surgical approach. There was a significant 2-year decrease in lordosis with the posterior approach group; however, all intervertebral angles were within typical ranges. Clinical outcomes were significantly improved 2 years postsurgery. There was no correlation between changes in lordosis and clinical outcomes. CONCLUSIONS: Interbody lumbar fusion with a threaded cylindrical cage does not appear to have any clinically relevant effects on segmental lordosis, which is maintained within anatomically normal levels. Clinical outcome measures show significant postsurgery improvement, and changes in lordotic angles are not predictive of clinical outcome. PMID- 11413427 TI - Does transverse apex coincide with coronal apex levels (regional or global) in adolescent idiopathic scoliosis? AB - STUDY DESIGN: Cross-sectional. OBJECTIVES: To identify the regional and global apexes of curves in adolescent idiopathic scoliosis and to compare the levels of those with the most rotated vertebral levels on computed tomography scans. SUMMARY OF BACKGROUND DATA: The terminology regarding the terms and definitions had been arbitrary until being refined and standardized by the Scoliosis Research Society Working Group on Three-Dimensional Terminology of Spinal Deformity. Apical vertebra or disc is defined as the most laterally deviated vertebra or disc in a scoliosis curve, but the most rotated vertebra (or disc) has not been included in this terminology. One study suggested that the most rotated vertebral level was always located at the apex. METHODS: Thirty-three structural curves of 25 consecutive patients scheduled for surgery for thoracic or thoracolumbar scoliosis were analyzed with standing anteroposterior radiographs and computed tomography scans covering the curve apexes and pelvis. Thoracic and lumbar curves were evaluated separately for all Type II curves. Vertebral rotations were normalized by the rotation of the pelvis. The most rotated vertebral (or disc) levels (transverse apex) were compared with the regional and global apex levels (vertebra or disc) (coronal apexes) of the corresponding curves separately. RESULTS: Regional and global apexes were at the same level in 18 (54.5%) curves, and within half a level in another 15 (45.4%), and the regional apex was one level higher in two curves (95% confidence levels: -0.82, +0.88). Comparison of the most rotated levels with regional and global apex levels revealed a higher variability, extending up to two levels for the global apex (95% confidence levels: -1.19, +1.54 levels for the global and -1.0, +1.41 levels for the regional apexes). CONCLUSION: This study demonstrated that the regional or global apex of a given curve is the most rotated level in only a minority of the curves. The most rotated level may be as far as two levels from the global apex and one level from the regional apex. PMID- 11413428 TI - The Effect of epsilon-aminocaproic acid on perioperative blood loss in patients with idiopathic scoliosis undergoing posterior spinal fusion: a preliminary prospective study. AB - STUDY DESIGN: A prospective study evaluating the efficacy of epsilon-aminocaproic acid (EACA) in decreasing perioperative blood loss in idiopathic scoliosis. OBJECTIVES: To compare the perioperative blood loss and need for blood replacement in two groups of patients undergoing surgery for idiopathic scoliosis. One group received intraoperative EACA; the other did not and served as controls. SUMMARY OF BACKGROUND DATA: Excessive blood loss increases the operative time, risk for blood product disease transmission, postoperative complications, and costs associated with posterior spinal fusion and instrumentation. EACA is an antifibrinolytic agent that has been shown to be effective in reducing perioperative blood loss during pediatric cardiothoracic surgical procedures. We hypothesized that it would also be effective in lowering blood loss during posterior spinal fusion for idiopathic scoliosis. METHODS: We compared the perioperative blood loss of 28 consecutive pediatric patients with idiopathic scoliosis who underwent posterior spinal fusion and received intraoperative EACA with 31 consecutive patients who did not receive this medication and served as a control group. RESULTS: The patients in both groups were similar. Patients in the EACA group demonstrated statistically significant decreases in total estimated perioperative blood loss and the need for autologous blood transfusion. The patients in the EACA group had no intraoperative or postoperative complications related to the use of this medication. CONCLUSIONS: Based on these preliminary findings, we believe that EACA is helpful in decreasing blood loss in patients undergoing posterior spinal fusion and instrumentation, and may decrease the number of autologous units needed to maintain safe perioperative hemoglobin levels, thereby improving safety and lowering cost associated with scoliosis surgery. PMID- 11413430 TI - The efficacy of antifibrinolytics in the reduction of blood loss during complex adult reconstructive spine surgery. AB - STUDY DESIGN: Controlled study to assess the efficacy of aprotinin and Amicar in reducing blood loss during complex spinal fusions. OBJECTIVES: To compare blood loss and the clotting profile with a thromboelastogram in patients with spinal deformities undergoing sequential anterior and posterior spinal fusions treated intraoperatively with either aprotinin or Amicar. SUMMARY OF BACKGROUND DATA: Spinal fusion for correction of adult spinal deformities is associated with large blood losses despite the implementation of multiple factors to reduce this blood loss. The antifibrinolytics aprotinin and Amicar have both been shown to reduce blood loss in other surgical procedures with the potential for large blood loss. Hence, we compared their efficacy for reducing blood loss in complex spinal fusions. METHODS: Sixty patients for elective sequential anteroposterior thoracolumbosacral fusions were randomly assigned to three groups: control, aprotinin, and Amicar. Patients were assessed for blood loss, transfusion requirements, postoperative complications, and coagulation profile using a thromboelastogram. RESULTS: The study demonstrated a significant reduction in total blood loss (aprotinin 3628 mL, Amicar 4056 mL, control 5181 mL) and transfusion requirements using the half-dose aprotinin regimen compared with Amicar or control. Aprotinin also preserved the thromboelastogram mean clot formation time, clot strength, and clotting index compared with Amicar or control. CONCLUSIONS: For complex spinal operations with large blood losses, the half-dose aprotinin regimen will reduce blood loss and the need for blood components and may have a role in reducing postoperative lung injury. PMID- 11413431 TI - The Longitudinal Assessment of Imaging and Disability of the Back (LAIDBack) Study: baseline data. AB - STUDY DESIGN: Prospective cohort study of randomly selected Veterans Affairs (VA) outpatients. OBJECTIVE: To determine the prevalence of magnetic resonance imaging (MRI) findings in the lumbar spine among persons without current low back pain or sciatica and to examine which findings are related to age or previous back symptoms. SUMMARY OF BACKGROUND INFORMATION: Previous studies of patients without low back pain have not explored the possible association of various MRI findings to past symptoms. METHODS: We randomly selected an age-stratified sample of subjects without low back pain in the past 4 months from clinics at a VA hospital. We collected information on demographics, comorbidity, functional status, and quality of life. MR images were obtained using a standardized protocol through each of the five lumbar disc levels. RESULTS: Of 148 subjects, 69 (46%) had never experienced low back pain. There were 123 subjects (83%) with moderate to severe desiccation of one or more discs, 95 (64%) with one or more bulging discs, and 83 (56%) with loss of disc height. Forty-eight subjects (32%) had at least one disc protrusion and 9 (6%) had one or more disc extrusions. CONCLUSION: Many MR imaging findings have a high prevalence in subjects without low back pain. These findings are therefore of limited diagnostic use. The less common findings of moderate or severe central stenosis, root compression, and extrusions are likely to be diagnostically and clinically relevant. PMID- 11413432 TI - Reliability and validity of the active straight leg raise test in posterior pelvic pain since pregnancy. AB - STUDY DESIGN: A cross-sectional analysis was performed in a group of women meeting strict criteria for posterior pelvic pain since pregnancy (PPPP). The scores on the Active Straight Leg Raise Test (ASLR test) were compared with the scores of healthy controls. OBJECTIVES: To develop a new diagnostic instrument for use in patients with PPPP. The objectives of the present study were to assess the validity and reliability of the ASLR test. SUMMARY OF BACKGROUND DATA: Various diagnostic tools are used to diagnose PPPP, but there is still a need for simple tests with high reliability, sensitivity, and specificity. METHODS: Reliability of the ASLR test was assessed in a group of 50 women with lumbopelvic pain of various etiologies and various degrees of severity. Sensitivity was assessed in 200 patients with PPPP and specificity in 50 healthy women. Sensitivity and specificity of the ASLR test were compared with the posterior pelvic pain provocation test (PPPP test). RESULTS: The test-retest reliability measured with Pearson's correlation coefficient between the two ASLR scores 1 week apart was 0.87. The intraclass correlation coefficient (ICC) was 0.83. Pearson's correlation coefficient between the scores of the patient and the scores of a blinded assessor was 0.78; the ICC was 0.77. In the patient group, the ASLR score ranged from 0-10; in the control group it ranged from 0-2. The best balance between specificity and sensitivity was found when scores 1-10 are designated as positive and zero as negative. With this cut-off point sensitivity of the test was 0.87 and specificity was 0.94. The sensitivity of the ASLR test is higher than the sensitivity of the PPPP test; an advantage of the ASLR test is the simplicity of measuring the score. CONCLUSION: The ASLR test is a suitable diagnostic instrument to discriminate between patients who are disabled by PPPP and healthy subjects. The test is easy to perform; reliability, sensitivity, and specificity are high. It seems that the integrity of the function to transfer loads between the lumbosacral spine and legs is tested by the ASLR test. PMID- 11413433 TI - Dynamic physiologic changes in lumbar CSF volume quantitatively measured by three dimensional fast spin-echo MRI. AB - STUDY DESIGN: Lumbar MRI of normal adults. Image analysis to measure lumbar CSF volumes at rest and during physiologic maneuvers. OBJECTIVES: 1) Validate an MRI technique to measure CSF volumes, 2) use this technique to measure the resting volume of lumbar CSF, 3) measure changes in CSF volume with physiologic maneuvers, and 4) demonstrate the anatomic basis for these volume changes. SUMMARY OF BACKGROUND DATA: Studies using radiograph and radionuclide myelography in dogs and humans in the 1960s-1980s qualitatively showed decreases in spinal CSF volume with physiologic maneuvers. Theories were proposed to explain these changes, but they could not be confirmed because only the contrast-laden CSF was visualized using these techniques. METHODS: Four adult volunteers had lumbar MRI using a fat-saturated T2-weighted three-dimensional fast spin-echo sequence. Quantitative analysis of images was used to measure lumbar CSF volume; the technique was validated using a water phantom. Lumbar CSF volume was measured 1) at rest, 2) with hyperventilation, 3) with abdominal compression, and 4) with both hyperventilation and abdominal compression. RESULTS: Resting lumbar CSF volume ranged from 28 to 42 mL. Reversible changes in lumbar CSF volume resulting from physiologic maneuvers are visualized by MR myelography and measured. The volume change (10% reduction in volume with hyperventilation, 28% with compression, and 41% with combined hyperventilation and abdominal compression) is directly visualized to be caused by engorgement of the epidural venous plexus, compressing the thecal sac. CONCLUSIONS: MRI provides a noninvasive means to measure spinal CSF volume and demonstrates the anatomic basis of physiologic volume changes. This has important implications for spinal anesthesia. PMID- 11413434 TI - Surgical and nonsurgical management of sciatica secondary to a lumbar disc herniation: five-year outcomes from the Maine Lumbar Spine Study. AB - STUDY DESIGN: A prospective cohort study. OBJECTIVE: To assess 5-year outcomes for patients with sciatica caused by a lumbar disc herniation treated surgically or nonsurgically. SUMMARY OF BACKGROUND DATA: There is limited knowledge about long-term treatment outcomes of sciatica caused by a lumbar disc herniation, particularly the relative benefits of surgical and conservative therapy in contemporary clinical practice. METHODS: Eligible, consenting patients recruited from the practices of orthopedic surgeons, neurosurgeons, and occupational medicine physicians throughout Maine had baseline interviews with mailed follow up questionnaires at 3, 6, and 12 months and annually thereafter. Clinical data were obtained at baseline from a physician questionnaire. Outcomes included patient-reported symptoms of leg and back pain, functional status, satisfaction, and employment and compensation status. RESULTS: Of 507 patients initially enrolled, 5-year outcomes were available for 402 (79.3%) patients: 220 (80%) treated surgically and 182 (78.4%) treated nonsurgically. Surgically treated patients had worse baseline symptoms and functional status than those initially treated nonsurgically. By 5 years 19% of surgical patients had undergone at least one additional lumbar spine operation, and 16% of nonsurgical patients had opted for at least one lumbar spine operation. Overall, patients treated initially with surgery reported better outcomes. At the 5-year follow-up, 70% of patients initially treated surgically reported improvement in their predominant symptom (back or leg pain) versus 56% of those initially treated nonsurgically (P < 0.001). Similarly, a larger proportion of surgical patients reported satisfaction with their current status (63% vs. 46%, P < 0.001). These differences persisted after adjustment for other determinants of outcome. The relative advantage of surgery was greatest early in follow-up and narrowed over 5 years. There was no difference in the proportion of patients receiving disability compensation at the 5-year follow-up. The least symptomatic patients at baseline did well regardless of initial treatment, although function improved more in the surgical group. CONCLUSIONS: For patients with moderate or severe sciatica, surgical treatment was associated with greater improvement than nonsurgical treatment at 5 years. However, patients treated surgically were as likely to be receiving disability compensation, and the relative benefit of surgery decreased over time. PMID- 11413436 TI - A new anterior cervical instrumentation system combining an intradiscal cage with an integrated plate: an early technical report. AB - STUDY DESIGN: To provide a technical report on a new device. Early outcome assessment of patients undergoing anterior cervical decompression and stabilization with the described technique. OBJECTIVES: To detail the operative and technical aspects of the new anterior cervical instrumentation and to identify its suitability for an extensive clinical study. SUMMARY OF BACKGROUND DATA: The controversy over the need for fusion in the surgery of cervical disc disease is in part fueled by the absence of an ideal technique. However, the design of the integrated anterior cervical plate and cage device (PCB) appears to provide immediate stability and to restore disc height and cervical lordosis in addition to reducing graft recipient and donor site-related complications. METHODS: After a standard anterior cervical discectomy and preparation of the disc space, the correct size of the cage with integrated plate is inserted. Self tapping screws are inserted without image intensifier. Cancellous bone chips are used. The technique was used in 29 patients that were followed prospectively from 5 to 24 months (median 8 months, interquartile range 6-15 months). RESULTS: The technique was easy to learn and implement. The authors' experience has been free of intraoperative complications. After surgery no screw backout or device failure was identified. Twenty-seven patients improved clinically. Donor site morbidity was trivial. No collars were applied after surgery. CONCLUSIONS: The design of the system appears to prevent bone-graft recipient site and donor site complications, provides immediate biomechanical stability, prevents screw backout or breakage, and restores posterior interbody height and lordosis. The device is of sufficient promise to warrant further close evaluation focusing on long-term outcome. The new radiolucent version has potential advantages for the assessment of fusion. PMID- 11413437 TI - Anterior cervical corpectomy and fusion using miniplate and screws in a 7-year old child with eosinophilic granuloma of the cervical spine. AB - STUDY DESIGN: This is a case report of a 7-year-old child with eosinophilic granuloma in the cervical spine, which underwent anterior cervical corpectomy and fusion by using Miniplate and screws. OBJECTIVES: To describe the use of Miniplate and screws for pediatric cervical anterior fusion. SUMMARY OF BACKGROUND DATA: Eosinophilic granuloma is a rare disease causing destructive bony lesions of the cervical spine in children. A complete resection and fusion were considered to be the preferable treatment in our case. However, cervical spinal fusion and instrumentation in children may be technically difficult because of the size of the vertebral body and the iliac bone. In addition, a proper device for an internal fixation in pediatric patients is not yet available. METHODS: A case of eosinophilic granuloma in pediatric spine was presented. RESULTS: We confirmed successful bony fusion and the restoration of the normal cervical curvature without recurrence of the tumor 2 years after the procedure. CONCLUSIONS: For proper internal fixation and prevention of dislodgement of the grafted bone, we used the Miniplate and screws as internal fixator after intralesional resection of the tumor mass. PMID- 11413438 TI - Image-guided anterior thoracolumbar corpectomy: a report of three cases. AB - STUDY DESIGN: A report of three cases of thoracolumbar vertebral collapse treated with image-guided anterior corpectomy. OBJECTIVES: To document the surgical technique and the usefulness of image-guided anterior corpectomy for thoracolumbar vertebral collapse. SUMMARY OF BACKGROUND DATA: Computer-assisted navigation systems can provide accurate three-dimensional surgical information intraoperatively. However, there is no clinical report regarding the application and usefulness of the computer-assisted navigation system for anterior thoracolumbar corpectomy. METHODS: After exposure of anterior and lateral aspects of the vertebral bodies through the transpleural approach, a reference frame was fixed to the thoracolumbar spinous process. Then thoracolumbar corpectomy and decompression were carried out under computer assistance. RESULTS: The tip of the standard probe and the angled rongeur could be monitored on three-dimensional images during surgery, and the retropulsed fragments within the spinal canal could be safely and completely removed under computer assistance. CONCLUSION: This image-guided procedure would aid surgeons in the complete and safe decompression of thoracolumbar injury. PMID- 11413439 TI - Elevated synthetic activity in the convex side of scoliotic intervertebral discs and endplates compared with normal tissues. AB - STUDY DESIGN: We measured concentrations of specific molecules reflecting matrix synthesis and degradation in normal and scoliotic intervertebral discs and endplates. OBJECTIVES: The aim of this work was to quantitate markers of matrix turnover in normal versus adolescent idiopathic scoliotic intervertebral discs and cartilaginous endplates. SUMMARY OF BACKGROUND DATA: Changes in the intervertebral disc and endplate composition have been implicated as possible etiologic factors in the pathogenesis of adolescent idiopathic scoliosis. To better understand this process, it is important to compare the turnover of matrix components in scoliotic and normal intervertebral disc and endplate tissues. This comparison may help to improve our understanding of the role that disc and endplate tissues may play in the induction and/or progression of idiopathic scoliosis. METHODS: Fifteen scoliotic and 17 normal intervertebral discs and endplates were analyzed for their water, collagen, proteoglycan, and protein content. In addition, newly synthesized aggrecan and collagen Types I and II were measured. Percent total denatured collagen was also determined. RESULTS: The total collagen content was significantly lower in the scoliotic anulus and endplate regions, whereas glycosaminoglycan (GAG) content was significantly lower in the scoliotic endplates and nucleus regions. Conversely, total protein content was significantly higher in scoliotic endplates and elevated in scoliotic nucleus regions. Water content was significantly lower in the scoliotic anulus and endplate regions. When comparing the concave and convex regions of scoliotic endplates, there was no significant difference in concentration of any matrix component. The major difference in the synthetic marker levels relates to the synthesis of Type II collagen, which was higher in the nucleus, anulus, and endplate regions of scoliotic discs than in the corresponding regions of normal tissues. By contrast, the percent total denatured collagen was significantly elevated in the nucleus of normal tissues compared with the scoliotic ones. CONCLUSIONS: The higher collagen Type II synthetic levels and increased total protein content with no matrix turnover suggest that scoliotic changes are due to an altered and ineffective synthetic response to a pathologic mechanical environment. PMID- 11413440 TI - A prospective randomized comparison of 270 degrees fusions to 360 degrees fusions (circumferential fusions). AB - STUDY DESIGN: Prospective randomized comparison of anterior lumbar interbody fusion (ALIF) plus transpedicular instrumentation plus posterolateral fusion (PLF) (360 degrees fusion) to ALIF plus transpedicular instrumentation without PLF (270 degrees fusion). OBJECTIVES: To compare the clinical outcomes, costs, and utilization of health resources of 360 degrees versus 270 degrees fusions. BACKGROUND: The 360 degrees fusion is effective, but its costs and utilization of health resources are high. The PLF often resorbs and may not be necessary. METHODS: Before and after surgery pain was measured by the Numerical Rating Scale (NRS), and function was measured by the Oswestry Low Back Disability Index (OSI). Costs were calculated by billing records. Operating times, blood loss, and hospital stays were measured at the time of hospital discharge. RESULTS: There were 48 patients: 21 women and 27 men. Mean age was 42 years. Follow-up averaged 35 months (range 24-45 months). In both 360 degrees and 270 degrees fusions, there were significant improvements in NRS and OSI, and the percentage of solid ALIF was high. Only 14% of PLF appeared solid bilaterally and 18% appeared solid on one side only. There were no significant differences in changes in NRS, changes in OSI, or percentage solid ALIF between the 360 degrees and 270 degrees fusions. However, the 270 degrees fusion group had significantly less blood loss, shorter operative times, shorter hospital stays, and lower professional fees, and although hospital charges were lower, this difference was not significant. CONCLUSION: Both the 360 degrees and 270 degrees fusions significantly reduce pain and improve function, and there are no significant clinical differences between them. However, there were shorter operating times, less blood loss, lower costs, and less utilization of health care resources associated with the 270 degrees fusions. PMID- 11413441 TI - Pigmented villonodular synovitis of lumbar facet joint: a rare cause of nerve root entrapment. AB - STUDY DESIGN: A case report of nerve root entrapment due to pigmented villonodular synovitis of lumbar facet joint is reported for which excision was performed. OBJECTIVE: To report a rare cause for nerve root claudication. SUMMARY OF BACKGROUND DATA: Pigmented villonodular synovitis is an uncommon synovial abnormality with an estimated incidence of 1.8 cases per million. Large joints, such as knee and hip, are commonly affected. Involvement of the facet joint is very rare. In our case a high index of suspicion from CT and MRI helped us in the appropriate management. METHODS: A 71-year-old man presented with severe back pain and right-sided L5 sciatica. CT and MRI scans showed a cystic lesion arising from the L5-S1 facet joint. Excision and adequate decompression in the form of undercutting facetectomy were done. RESULTS: The patient had symptomatic improvement with surgery, and at the 3-year follow-up he showed no signs of recurrence. CONCLUSION: Unless pigmented villonodular synovitis is considered in the differential diagnosis of tumors of the vertebral column causing nerve root claudication, it may be overlooked. PMID- 11413442 TI - Giant intrasacral cellular schwannoma treated with high sacral amputation. AB - STUDY DESIGN: A case report of a man with a gigantic cellular schwannoma in the sacrum treated with high sacral amputation accompanied by careful nerve root sparing dissection. OBJECTIVES: To describe the atypical clinical course of an intrasacral cellular schwannoma and the surgical procedure of high sacral amputation performed in a way to prevent needless sacrifice of functionally essential nerve roots. SUMMARY OF BACKGROUND DATA: Fundamentally, a cellular schwannoma is a benign tumor, but the clinical course is atypical. The symptoms are mild and the clinicopathologic features often mislead us to make a diagnosis of malignancy. The occurrence rate of intraosseous cellular schwannoma was reported to be 0.2% of all bony tumors, and the main location was the retroperitoneal space in the pelvis. Forty-one cases of giant intrasacral schwannomas have been reported so far. Among them, large sacral schwannoma with anterior cortex erosion and associated intrapelvic extension was extremely rare. METHODS: The patient presented with a 5-year history of right leg and buttock pain, which did not disturb his daily activities. After a histopathologic diagnosis and a complete set of image studies, high sacral amputation with preservation of uninvolved nerve roots was performed at S1-S2 through a combined anterior and posterior approach. Both S1 nerve roots and the right S2-S3 nerve roots were saved using a threaded saw. The lumbar spine was stabilized to the pelvic girdle using spinal instrumentation with posterolateral fusion. RESULTS: Eighteen months after the tumor was resected the patient had a very good clinical outcome, and there were no radiologic signs of instability or recurrence of the tumor. Locomotor function of both lower extremities and bowel and urinary functions were well maintained. The patient returned to his previous work. CONCLUSIONS: High sacral amputation following a combined anteroposterior approach provided good results without causing any disability. A detailed preoperative planning and careful dissection of uninvolved nerve roots prevented unnecessary neurologic impairment in locomotion and the detrusor and anorectal function. PMID- 11413443 TI - Cervical spondylotic amyotrophy with intramedullary cavity formation. AB - STUDY DESIGN: A case report. OBJECTIVE: To show that an intramedullary lesion was the cause of cervical spondylotic amyotrophy. SUMMARY OF BACKGROUND DATA: Cervical spondylotic amyotrophy is the clinical syndrome characterized by muscle wasting and weakness in the upper extremities without a remarkable sensory loss or spastic tetraparesis. It is still unclear whether the ventral roots or the anterior horn are selectively damaged. METHODS: Magnetic resonance imaging and delayed computed tomographic myelography were performed on a patient who showed severe wasting of the left triceps muscle without any sensory disturbance or long tract sign. RESULTS: On sagittal magnetic resonance images, a linear area was noted within the spinal cord at C6 and C7 spinal segments, which showed low signal intensity on T1-weighted image and high signal intensity on T2-weighted image. On axial T1-weighted image intramedullary low signal intensity area was observed, which was located in the left anterior horn. On axial T2-weighted image the area showed high signal intensity. A delayed (6 hours) computed tomographic scan after intrathecal injection of metrizamide revealed intramedullary enhancement in the area corresponding to the left anterior horn, which would represent cavitation or cystic necrosis. CONCLUSIONS: This is the first case report of cervical spondylotic amyotrophy, in which intramedullary lesion was confirmed only at the affected side of the spinal cord. PMID- 11413446 TI - Cell division: Not all are born equal. PMID- 11413448 TI - Plant development: Stomatal waxing and waning. PMID- 11413449 TI - Protein degradation: Parkin finds a partner and a victim. PMID- 11413450 TI - Meiosis: Synapsis spoilt. PMID- 11413451 TI - Protein-protein interactions: Phosphothreonine lego. PMID- 11413452 TI - Cell cycle: Two's company. PMID- 11413453 TI - Apoptosis: Bax to Bak. PMID- 11413454 TI - Cytoskeleton: Molecular chauffeurs. PMID- 11413457 TI - Cell division: Dodging death at division? PMID- 11413458 TI - Technique: Fluorescent timer. PMID- 11413459 TI - Membrane dynamics: Variation on a theme. PMID- 11413461 TI - Asymmetric cell division during animal development. AB - Although most cells produce two equal daughters during mitosis, some can divide asymmetrically by segregating protein determinants into one of their two daughter cells. Interesting parallels exist between such asymmetric divisions and the polarity established in epithelial cells, and heterotrimeric G proteins might connect these aspects of cell polarity. The discovery of asymmetrically segregating proteins in vertebrates indicates that the results obtained in invertebrate model organisms might also apply to mammalian stem cells. PMID- 11413462 TI - Mitotic kinases as regulators of cell division and its checkpoints. AB - Mitosis and cytokinesis are undoubtedly the most spectacular parts of the cell cycle. Errors in the choreography of these processes can lead to aneuploidy or genetic instability, fostering cell death or disease. Here, I give an overview of the many mitotic kinases that regulate cell division and the fidelity of chromosome transmission. PMID- 11413463 TI - Plant cell division: building walls in the right places. AB - Plant cells are surrounded by walls that define their shapes and fix their positions with tissues. Consequently, establishment of a plant's cellular framework during development depends largely on the positions in which new walls are formed during cytokinesis. Experiments using various approaches are now building on classical studies to shed light on the mechanisms underlying the spatial control of cytokinesis. PMID- 11413464 TI - Microtubule-associated proteins in plants--why we need a MAP. AB - Plants have four main microtubule assemblies. Three are involved in arranging when and where the cell wall is laid down and have no direct homologues in animals. Microtubule-associated proteins are important components of these assemblies, and we are now starting to uncover what these proteins are and how they might work. PMID- 11413465 TI - Not being the wrong size. AB - Size regulation is a never-ending problem. Many of us worry that parts of ourselves are too big whereas other parts are too small. How organisms--and their tissues--are programmed to be a specific size, how this size is maintained, and what might cause something to become the wrong size, are key problems in developmental biology. But what are the mechanisms that regulate the size of multicellular structures? PMID- 11413466 TI - Protein interaction maps for model organisms. PMID- 11413467 TI - Breaking the mitochondrial barrier. AB - Pro- and anti-apoptotic members of the Bcl-2 family control the permeability of the outer mitochondrial membrane. They could do this either by forming autonomous pores in the membrane or by collaborating with components of the permeability transition pore. Here we discuss why we favour the first of these possibilities. PMID- 11413468 TI - The mitochondrion in apoptosis: how Pandora's box opens. AB - There is widespread agreement that mitochondria have a function in apoptosis, but the mechanisms behind their involvement remain controversial. Here we suggest that opening of a multiprotein complex called the mitochondrial permeability transition pore complex is sufficient (and, usually, necessary) for triggering apoptosis. PMID- 11413469 TI - Walther Flemming: pioneer of mitosis research. AB - The German anatomist Walther Flemming began his pioneering studies of mitosis almost 150 years ago. What were his achievements, and where have his discoveries led? PMID- 11413472 TI - A cell cycle controller rewrites its CV. PMID- 11413475 TI - SNARE hypothesis 2000. PMID- 11413476 TI - Another star on drugs. PMID- 11413478 TI - From duckweed to DNA... PMID- 11413479 TI - . . . and from repair to remodelling. PMID- 11413482 TI - DIABLO is double trouble. PMID- 11413483 TI - Caps and cancer. PMID- 11413484 TI - Pocket the difference. PMID- 11413485 TI - The versatility and universality of calcium signalling. AB - The universality of calcium as an intracellular messenger depends on its enormous versatility. Cells have a calcium signalling toolkit with many components that can be mixed and matched to create a wide range of spatial and temporal signals. This versatility is exploited to control processes as diverse as fertilization, proliferation, development, learning and memory, contraction and secretion, and must be accomplished within the context of calcium being highly toxic. Exceeding its normal spatial and temporal boundaries can result in cell death through both necrosis and apoptosis. PMID- 11413486 TI - A tale of toroids in DNA metabolism. AB - A strikingly large number of the proteins involved in DNA metabolism adopt a toroidal -- or ring-shaped -- quaternary structure, even though they have completely unrelated functions. Given that these proteins all use DNA as a substrate, their convergence to one shape is probably not a coincidence. Ring forming proteins may have been selected during evolution for advantages conferred by the toroidal shape on their interactions with DNA. PMID- 11413487 TI - Lipid rafts and signal transduction. AB - Signal transduction is initiated by complex protein-protein interactions between ligands, receptors and kinases, to name only a few. It is now becoming clear that lipid micro-environments on the cell surface -- known as lipid rafts -- also take part in this process. Lipid rafts containing a given set of proteins can change their size and composition in response to intra- or extracellular stimuli. This favours specific protein-protein interactions, resulting in the activation of signalling cascades. PMID- 11413488 TI - New targets for inhibitors of HIV-1 replication. AB - Despite the success of protease and reverse transcriptase inhibitors, new drugs to suppress HIV-1 replication are still needed. Several other early events in the viral life cycle (stages before the viral genome is inserted into host cell DNA) are susceptible to drugs, including virus attachment to target cells, membrane fusion and post-entry events such as integration, accessory-gene function and assembly of viral particles. Among these, inhibitors of virus-cell fusion and integration are the most promising candidates. PMID- 11413489 TI - Walking on two heads: the many talents of kinesin. AB - The gallop of a race horse and the minute excursions of a cellular vesicle have one thing in common: they are based on the directional movement of proteins termed molecular motors -- many trillions in the case of the horse, just a few in the case of the cell vesicle. These tiny machines take nanometre steps on a millisecond timescale to drive all biological movements. Over the past 15 years new biochemical and biophysical approaches have allowed us to take a giant step forward in understanding the molecular basis of motor mechanics. PMID- 11413490 TI - Multilevel regulation of the circadian clock. AB - Living organisms adapt to light-dark rhythmicity using a complex programme based on internal clocks. These circadian clocks, which are regulated by the environment, direct various physiological functions. As the molecular mechanisms that govern clock function are unravelled, we are starting to appreciate simple patterns as well as exquisite layers of regulation. PMID- 11413491 TI - The past, present and future of molecular computing. AB - Ever since scientists discovered that conventional silicon-based computers have an upper limit in terms of speed, they have been searching for alternative media with which to solve computational problems. That search has led them, among other places, to DNA. PMID- 11413492 TI - Hayflick, his limit, and cellular ageing. AB - Almost 40 years ago, Leonard Hayflick discovered that cultured normal human cells have limited capacity to divide, after which they become senescent -- a phenomenon now known as the 'Hayflick limit'. Hayflick's findings were strongly challenged at the time, and continue to be questioned in a few circles, but his achievements have enabled others to make considerable progress towards understanding and manipulating the molecular mechanisms of ageing. PMID- 11413493 TI - Cancer: looking outside the genome. PMID- 11413495 TI - Direct evidence for bioconversion of vitamin E acetate into vitamin E: an ex vivo study in viable human skin. AB - For better stability, vitamin E is commonly used as the non-active esterified pro drug. Such esters are postulated to be hydrolyzed to the free active form by skin related esterases. So far, successful conversion of esterified vitamin E to free vitamin E (tocopherol) has been mainly delineated from observed biological effects. Quantitative evidence in human skin is poor. In vitro and in vivo studies on human and animal skin have proved ambiguous. Formulation-based effects may have added to this controversy. In the present study, comparable amounts of vitamin E acetate (i) in oil (Mygliol-812N), (ii) surfactant-solubilized in water, (iii) encapsulated in liposomes, or (iv) encapsulated in Nanotopes were applied to human skin mounted in modified Franz-perfusion chambers that permit emulation of both open or occlusive conditions. The distribution of vitamin E(total) (vitamin E acetate + vitamin E) was assessed on the skin surface, in the horny layers, and in the underlying skin by high-pressure liquid chromatography (HPLC), with a recovery higher than 90%. Vitamin E acetate in Mygliol deposited exclusively on the surface and in the stratum corneum. In contrast, solubilized or encapsulated vitamin E acetate deposited also in the underlying skin. Nanotopes performed best, followed by liposomes and solubilized vitamin E acetate. Non-occlusive application favored deposition in the skin relative to occlusive application. Conversion of vitamin E acetate to vitamin E was not observed on the skin surface or in the horny layers, while in the underlying skin up to 50% of the vitamin E(total) was deacetylated. PMID- 11413496 TI - Abrasiveness evaluation of silica and calcium carbonate used in the production of dentifrices. AB - Our purpose was to apply a radiometric method to an abrasiveness evaluation in samples of silica and calcium carbonate used as an abrasive in a dentifrice, to help in a prudent selection of materials by dentifrice producers. The results of RDA (radioactive dentin abrasion) abrasiveness indices obtained for these compounds varied from 136 to 19. The relative standard deviations of these RDA results varied from 5.9% to 11.8%, showing a good precision in the method. Also, the results obtained indicated that the abrasiveness indices increase with the particle size of the material. A comparison between different abrasives with similar particle sizes showed that silica presents higher RDA values than calcium carbonate. PMID- 11413497 TI - Secondary ion mass spectrometric investigation of penetration of coconut and mineral oils into human hair fibers: relevance to hair damage. AB - An attempt has been made to show the difference in the penetrability of coconut oil and mineral oil in human hair. We have used secondary ion mass spectrometry (SIMS) in combination with a time-of-flight (TOF) mass spectrometer. Characteristic ions formed by the pure components when bombarded with gallium ions have been identified with their m/z values. The distribution of the ion, characteristic of the particular treatment, has been established in the cross sections of hair treated with coconut and mineral oils. The results show that coconut oil penetrates the hair shaft while mineral oil does not. The difference may be due to the polarity of the coconut oil compared to the nonpolar nature of the mineral oil. The affinity of the penetrant to the protein seems to be the cause for this difference in their behavior. This study also indicates that the swelling of hair is limited by the presence oil. Since the process of swelling and deswelling of hair is one of the causes of hair damage by hygral fatigue, coconut oil, which is a better penetrant than mineral oil, may provide better protection from damage by hygral fatigue. PMID- 11413498 TI - Mechanical extension of human hair and the movement of the cuticle. AB - Goniophotometry has enabled measurement of the angle that the surface of the cuticle (the scales) of a human hair fiber makes with the axis of the hair shaft. This measuring technique has been used to obtain the change of this scale angle with extension of a hair fiber under fixed conditions of temperature and relative humidity. Based on a simple model of overlapping scales of the hair cuticle, analysis shows that, for hair fibers extended to strains above 10% at 35% relative humidity and at 35 degrees C, overlapping scales become progressively detached from each other. This scale detachment has been suggested to result from the mechanical failure of the endocuticle layer in the scale structure. This endocuticle layer is low in disulfide cross-linking, which would result in a lowered stiffness and greater extensibility, as indicated by the higher swelling of the layer in water as against the highly cross-linked exocuticle layers. The greater extensibility of the endocuticle would also explain the greater distortion of this layer under stress, but it would not follow that endocuticular failure under stress should result. An alternative mechanism of failure of adhesion between overlapping scales in the cuticle is suggested, based on the involvement of the hydrophobic upper-beta-layer with its surface of 18 methyleicosanoic acid (18-MEA), which may provide mobility and a reduction in adhesion between scales. This potential failure of the cementing of the overlapping scale structure due to the 18-MEA is discussed, with particular reference to the standard permanent setting procedure of human hair. PMID- 11413500 TI - New requirements for marriage license? PMID- 11413501 TI - Self-harm behavior and no-suicide contracting in psychiatric inpatient settings. AB - Although negotiation of no-suicide contracts is common practice, research regarding the outcomes of contracting is inadequate. The purpose of this retrospective review of medical records was to examine how no-suicide contracting affected the likelihood of self-harm behavior in psychiatric inpatient settings. Thirty-one patients (4.8%) engaged in self-harm behaviors representing 2.64% of all patients admitted to both study settings during the 6 (1/2)-month period from which data were collected. Approximately half of those patients expressed suicidal intent. Logistic regression analysis suggested that patients with no suicide contracts and with higher levels of restriction had a significantly higher likelihood of self-harm behavior (OR = 7.43 and 2.47, respectively, p = .005). Consistency of nursing assignment is likely associated with a lower probability of self-harm (OR = .07) but p.068 when this variable was included in the model. Prevention of self-harm behaviors by the use of no-suicide contracting is not shown. Negotiation of a contract is likely a reflection of staff assessment that the patient was at high risk for suicide. These findings confirm the need for thorough, ongoing assessment of suicidal risk, whether or not a patient has agreed to a no-suicide contract. PMID- 11413502 TI - The role of advanced practice psychiatric nurses in treating tobacco use and dependence. AB - Tobacco use is treatable and the benefits of smoking cessation are impressive, yet like other health care providers, psychiatric nurses have failed to consistently assess and treat tobacco use effectively. Tobacco use continues to cause illness, disability, and death at unprecedented rates. Nicotine addiction is drug abuse and it is a chronic disease and needs to be treated as such. There are clear health benefits to smoking cessation regardless of the age of smoking initiation or the age of smoking cessation. Advanced practice psychiatric nurses (APPNs) are in a unique position to make an impact on a smoker's risk of suffering from tobacco-related diseases: the treatment of choice is the combination of pharmacotherapy and psychosocial interventions, both within the realm of APPN practice; psychiatric and substance abusing patients consistently demonstrate increased rates of cigarette smoking compared with healthy controls; and tobacco use is drug abuse, a treatable chronic disease. The ability of APPNs to deliver psychotherapeutic and pharmacologic care secures a position in the forefront of treating tobacco use. In addition to intervention, psychiatric nurses need to step up and take an active role in initiating and supporting tobacco control policy and legislation. PMID- 11413503 TI - Smooth a heart with water: Thai mothers care for a child with schizophrenia. AB - Little is known about caregiving of persons with schizophrenia in Asian families. This report explores culturally specific beliefs and patterns of response among Thai families. In-depth interviews were conducted with 12 Thai mothers of adult children with schizophrenia. Thai mothers responded to children's symptoms by practicing Thum-jai and creating a calm family environment. Maternal response was constituted by concerns about social expectations, escalation of the symptoms, and violating cultural values. Assessment of and attunement to Buddhist beliefs in the care of Thai families and immigrants are recommended. PMID- 11413504 TI - A phenomenological exploration of loneliness in the older adult. AB - Loneliness is an important phenomenon of concern for psychiatric mental health nursing. Research findings have related loneliness in older adults to mental health problems especially depression. However, little is known about loneliness from the perspective of older adults living in the community. A phenomenological inquiry using Giorgi's research methodology was undertaken to explore and describe the meaning of loneliness for older adults. Twenty participants, most who are over the age of 75, were interviewed and 1385 naive meaning units were extrapolated from tape-recorded verbatim transcripts. These units were clustered into five major themes describing the loneliness experience. The findings are described and discussed relative to the literature on loneliness. Implications for mental health nursing are presented. PMID- 11413505 TI - Empathy from a nursing perspective: Moving beyond borrowed theory. AB - Empathy is a concept deeply rooted in and central to professional nursing. Although viewed as an important concept, little consensus exists in the professional literature about either the definition or the application of the concept to nursing practice. This article will compare two theories of empathy, one borrowed from Kohut's self-psychology model and one derived from King's Interacting Systems nursing framework. The two theories are examined to clarify issues pertaining to the concept of empathy and to identify the contributions and limitations of borrowed theory as the basis for nursing practice. The article explores similarities and difference in the two views of empathy and highlights the necessity of developing nursing science from theory based in nursing which reflects the very unique nature of nursing practice. Nursing as a profession is distinct and unique, and borrowed theory must be questioned for its fit and applicability to the profession. The article concludes that empathy is a nursing phenomenon needing to be studied from a nursing perspective. The results of continued use of borrowed theory pertaining to empathy is discussed including the delay and misdirection of ongoing activity to develop the concept of empathy. PMID- 11413506 TI - New directions for feminist therapy based on social constructionism. AB - Feminist therapy has made significant contributions in the area of women's mental health care. Of late, however, critics have argued that feminist therapists are neglecting the needs of many women. The unique perspectives of women of color, lower and upper class women, lesbians, and other persons have been ignored. As such, it is proposed that social constructionism offers a metaframework for reinterpreting feminist therapy tenets to better address the needs of a broad range of individuals. Clinical implications are offered along with future directions for research and education. PMID- 11413507 TI - Rapid engraftment after allogeneic transplantation of density-enriched peripheral blood CD34+ cells in patients with advanced hematologic malignancies. AB - BACKGROUND: Acute graft versus host disease (GVHD) remains a major cause of morbidity and mortality after allogeneic hematopoietic cell transplantation. Preclinical studies have suggested that a T-cell subset with a CD4-/CD8- double negative (DN) T-cell phenotype is capable of suppressing GVHD. Double-negative T cells can be mobilized into the peripheral blood with granulocyte colony stimulating factor (G-CSF) and enriched by density centrifugation. The current study was performed to study the feasibility and safety of applying a density gradient separation technique for enrichment of CD34+ and DN T cells, while depleting CD4+ and CD8+ single-positive (SP) T cells from peripheral blood progenitor cells (PBPCs) for the purpose of allogeneic transplantation. METHODS: Twenty-five patients with advanced hematologic malignancies were treated with a myeloablative preparative regimen consisting of fractionated total body irradiation, etoposide, and cyclophosphamide. Human leukocyte antigen identical donors were mobilized with G-CSF PBPC collected by apheresis. The apheresis product was applied to a single-step density gradient, and the low-density cell population was collected. The low-density cell population was infused as the sole source of allogeneic cells after myeloablative therapy. Graft versus host disease prophylaxis consisted of cyclosporine with or without prednisone. RESULTS: CD34 cell recovery was efficient with a median 72% yield, providing for a median CD34+ cell dose of 6.5 x 10(6)/kg (range,1.0- 13.9 x 10(6)/kg). CD3+CD4+ or CD3+CD8+ SP T cells were depleted by a median of 94.4% (range, 58.8- 99.2%), and the ratio of CD34+:SP T cells increased 10-fold. Double-negative T cells were depleted by 92% (range, 18.8- 99.4%), thus the ratio of DN:SP T cells increased less than 2-fold in 71% of apheresis samples tested. Hematopoietic engraftment was rapid, and there was no occurrence of graft failure in examinable patients. Median time to absolute neutrophil count greater than 0.5 x 10(9)/L and platelet count greater than 20 x 10(9)/L was 10.5 and 12 days, respectively. The incidence of Grade 2-4 acute GVHD was 26% (95% confidence interval [CI], 6-45%), although not all patients were examinable due to an unexpectedly high nonrecurrence mortality that at Day 180 was 62% (95% CI, 40-83%). CONCLUSIONS: These data suggest that T-cell subset manipulation via density gradient separation is a safe procedure and allowed rapid hematopoietic recovery. Selective enrichment of a donor DN T-cell subset was observed in only a few and was not associated with a reduced incidence of GVHD. However, the low-density selected cells still resulted in GVHD, and there was a high treatment-related mortality. PMID- 11413508 TI - Anemia as an independent prognostic factor for survival in patients with cancer: a systemic, quantitative review. AB - BACKGROUND: Anemia is common in cancer patients, although the prevalence is influenced both by the type of malignancy and the choice of treatment. Individual studies have compared the survival of patients with and without anemia and have shown reduced survival times in patients with various malignancies, including carcinoma of the lung, cervix, head and neck, prostate, lymphoma, and multiple myeloma. The objective of this study was to systematically review, to summarize, and to obtain an overall estimate of the effect of anemia on survival in patients with malignant disease. METHODS: A comprehensive literature review was carried out using the MEDLINE data base and reviewing the reference lists from published studies. Two hundred papers were identified. Of these, 60 papers that reported the survival of cancer patients according to either hemoglobin levels or the presence of anemia were included. Among these papers, 25% related to patients with lung carcinoma, 17% related to patients with head and neck carcinoma, 12% related to patients with multiple myeloma, 10% related to patients with prostate carcinoma, 8% related to patients with cervicouterine carcinoma, 7% related to patients with leukemia, 5% related to patients with lymphoma, and 16% related to patients with other types of malignancies. RESULTS: The relative risk of death increased by 19% (95% confidence interval, 10-29%) in anemic patients with lung carcinoma, by 75% (37-123%) in anemic patients with head and neck carcinoma, by 47% (21-78%) in anemic patients with prostate carcinoma, and by 67% (30-113%) in anemic patients with lymphoma. The overall estimate increase in risk was 65% (54 77%). CONCLUSIONS: Anemia is associated with shorter survival times for patients with lung carcinoma, cervicouterine carcinoma, head and neck carcinoma, prostate carcinoma, lymphoma, and multiple myeloma. PMID- 11413509 TI - Methylation of p15 and p16 genes in adult acute leukemia: lack of prognostic significance. AB - BACKGROUND: Gene-promoter methylation is an epigenetic mechanism of transcription inactivation. In this study, the authors investigated the frequency and prognostic significance of p15 and p16 gene methylation in adult acute leukemia. METHOD: The methylation-specific polymerase chain reaction (MS-PCR) was used to analyze p15 and p16 gene methylation in 49 cases of acute lymphoblastic leukemia (ALL) and 29 cases of acute myelogenous leukemia (AML). RESULTS: At presentation, 93 % of cases of AML (8 of 8 M1, 10 of 11 M2, 2 of 2 M4, 5 of 6 M5, and 2 of 2 M6; French-American-British classification system) showed p15 methylation, but none showed p16 methylation. In ALL, 57% (5 of 8 T-ALL, 16 of 30 common-ALL, 6 of 7 pre-B ALL, and 1 of 4 early B-precursor ALL) showed p15 methylation. Only 6% showed p16 methylation, all of whom had concomitant p15 methylation. One patient acquired p16 methylation during relapse. In 23 ALL karyotyped cases, p15 methylation was found in 6 of 9 cases with normal karyotype, 3 of 7 cases with the Philadelphia chromosome, 3 of 5 cases with complex, 1 with hyperdiploidy, and 1 with trisomy 21. Three more cases with unsuccessful karyotyping but bcr/abl fusion showed p15 methylation as well. Five ALL patients were tested serially for minimal residual disease (MRD) with MS-PCR that has a sensitivity of 10(-4) to 10(-5). All showed continuous positive MS-PCR that heralded hematologic relapse. The prognostic significance of p15 methylation was tested in ALL patients, showing no impact on complete remission, 5-year overall survival, or 5-year disease-free survival. CONCLUSION: Gene methylation of p15, but not p16, is frequent in adult acute leukemias. Methylation of p15 at diagnosis was of no prognostic significance in ALL but may be useful for monitoring MRD. PMID- 11413510 TI - Dynamic magnetic resonance imaging of regional contrast access as an additional prognostic factor in pediatric osteosarcoma. AB - BACKGROUND: The purpose of this article was to evaluate the utility of a pharmacokinetically modeled measure of regional contrast access, based on dynamic contrast-enhanced magnetic resonance imaging (MRI) studies after preoperative chemotherapy, as a predictor of disease free survival in osteosarcoma. METHODS: The kinetic parameters of a two-compartment pharmacokinetic model of MRI contrast agent accumulation were analyzed in relation to disease free survival in 31 patients who received protocol-based therapy for nonmetastatic osteosarcoma of the extremities. The modeled exchange rate of contrast between the plasma and the tumor extravascular extracellular fluid space served as a measure of regional contrast access. The prognostic impact of both the clinically accepted standard of histologic evaluation of tumor necrosis and the regional contrast access were analyzed with tumor size as an influential factor. RESULTS: Although the histologic grade of response was not a statistically significant prognostic factor in these patients (P = 0.884), regional contrast access after preoperative chemotherapy was significantly predictive of disease free survival (P = 0.035) in the Cox proportional hazards model. Lower regional access before surgery and smaller tumor size were associated with a better treatment outcome. Log-rank analyses of Kaplan-Meier curves indicated that the impact of regional access was most pronounced in patients with larger tumors (P = 0.052). Higher regional access at presentation also was associated significantly with greater decreases during therapy. CONCLUSIONS: Dynamic MRI estimates of regional contrast access after preoperative chemotherapy, when combined with tumor size, holds promise for the early identification of patients at risk of recurrence. The availability of such response predictors could facilitate the development of risk-adapted treatment approaches. PMID- 11413511 TI - Low bone mineral density in hormone-naive men with prostate carcinoma. AB - BACKGROUND: The objective of this study was to determine the prevalence of low bone mineral density in men with prostate carcinoma and no history of androgen deprivation therapy. METHODS: The authors conducted a cross-sectional study in 41 hormone-naive men with locally advanced, lymph node positive, or recurrent prostate carcinoma and no radiographic evidence of bone metastases. Bone mineral density of the total hip, posterior-anterior (PA) lumbar spine, and lateral lumbar spine was determined by dual-energy X-ray absorptiometry (DXA) using a densitometer. Trabecular bone mineral density of the lumbar spine was determined by quantitative computed tomography (QCT). Bone mineral density results were expressed in standard deviation units relative to young adult men (T score) and relative to age-matched men (Z score). RESULTS: Fourteen of 41 men (34%; 95% confidence interval [95% CI], 20-51%) had T scores < -1.0 at one or more skeletal sites by DXA, 12 of 41 men (29%; 95% CI, 16-42%) had T scores between -1.0 and 2.5, and 2 of 41 men (5%; 95% CI, 1-17%) had T scores < -2.5. Thirty-nine of 41 men (95%; 95% CI, 83-99%) had T scores < -1.0 by QCT, 13 of 41 men (31%; 95% CI 18-48%) had T scores between -1.0 and -2.5, and 26 of 41 men (63%; 95% CI, 47 78%) had T scores < -2.5. T scores for trabecular bone mineral density of the lumbar spine were significantly lower than T scores for either the total hip (P < 0.001) or the PA lumbar spine (P < 0.001). The mean Z score for trabecular bone mineral density of the lumbar spine was -0.7 +/- 0.9. Hypogonadism, hypovitaminosis D, and dietary calcium intakes below the Recommended Daily Allowance were observed in 20%, and 17%, and 59% of study participants, respectively. CONCLUSIONS: Many hormone-naive men with prostate carcinoma have low bone mineral density. QCT is a more sensitive method than DXA for diagnosing low bone mineral density in this patient population. Trabecular bone mineral density is lower than expected for age and risk factors for osteoporosis are common. PMID- 11413512 TI - Analysis of the prognostic effects of inclusion in a clinical trial and of myelosuppression on survival after adjuvant chemotherapy for breast carcinoma. AB - BACKGROUND: Treatment-related factors that might influence survival after adjuvant treatment for breast carcinoma include treatment as part of a clinical trial, toxicity of treatment, the regimen and schedule used, and dose intensity. METHODS: The authors reviewed the records of 680 patients with breast carcinoma who received adjuvant treatment with cyclophosphamide, methotrexate, and 5 fluorouracil (CMF) at Princess Margaret Hospital between 1980-1990. They analyzed the effects on survival of inclusion in a clinical trial (n = 160) and of experiencing Grade 3/4 myelosuppression (according to National Cancer Institute Common Toxicity Criteria) (n = 227, with available data for 584 patients). In an exploratory analysis, the authors examined the effects of the CMF regimen ("classic" CMF [n = 417] vs. intravenous CMF [n = 243]) and of relative dose intensity. Each of these factors was tested in a Cox proportional hazards model with the known prognostic factors of N classification, T classification, estrogen receptor (ER) and progesterone receptor (PR) status, and use of adjuvant hormonal therapy to determine whether they provided additional prognostic information. RESULTS: In univariate analysis, inclusion in a clinical trial was associated with better survival (P = 0.02) with a nonsignificant trend when corrected for other prognostic factors in a multivariate analysis (hazard ratio [HR] = 0.77; 95% confidence interval [CI], 0.56-1.04). There was a similar trend for patients experiencing myelosuppression (HR = 0.77; 95% CI, 0.59-1.00). In exploratory analysis the use of classic CMF, with higher absolute dose intensity, also was associated with a trend toward improved survival (HR = 0.79; 95% CI, 0.63-1.00). CONCLUSIONS: The results of the current study suffer from the inherent problems of retrospective analysis, but, similar to findings for other disease sites, they suggest that patients included in clinical trials have better outcome. Classic CMF should be used when this regimen is selected for adjuvant treatment, and dose adjustment resulting in moderate myelosuppression should be explored in future clinical trials. PMID- 11413513 TI - Prognostic significance of the number of lymph nodes examined in patients with lymph node-negative breast carcinoma. AB - BACKGROUND: A recent report suggested that the number of lymph nodes examined was a strong predictor of survival in patients with lymph node-negative breast carcinoma. Among women who had >or= 20 lymph nodes examined, the risk of dying from breast carcinoma within 5 years was increased nearly 4-fold compared with women who had fewer lymph nodes examined. Because these findings were based on a relatively small cohort of patients, corroborative studies with larger patient populations were needed. METHODS: The authors studied the relation between the number of lymph nodes examined and breast carcinoma survival among 911 women with lymph node-negative breast carcinoma with a median length of follow-up of 84 months. The association between other prognostic indicators and survival and the number of lymph nodes examined also was investigated. RESULTS: The number of lymph nodes examined was not found to be associated with either 5-year or long term survival. The proportion of women dying from breast carcinoma was the same (8%) in both groups (those patients with >or= 20 lymph nodes examined vs. those in whom < 20 lymph nodes were examined) and the hazard ratio was 0.98 (95% confidence interval, 0.58-1.64). CONCLUSIONS: In this larger study population, the authors failed to confirm the findings of an earlier investigation in which having a larger number of lymph nodes examined was associated with poorer survival. This finding suggests that it is unlikely the number of lymph nodes examined is an important prognostic indicator in patients with lymph node negative breast carcinoma. PMID- 11413514 TI - Computer-assisted complete three-dimensional reconstruction of the mammary ductal/lobular systems: implications of ductal anastomoses for breast-conserving surgery. AB - BACKGROUND: The intraductal spread of breast carcinoma can occur along the mammary ductal/lobular systems (MDLS) with no invasion of tissues. Because ductal anastomoses in the MDLS are considered to be a possible risk factor for extensive intraductal spread of breast carcinoma, the architecture of the MDLS has important therapeutic implications for patients treated with breast-conserving surgery. METHODS: An entire breast resected by subcutaneous mastectomy from a 69 year-old woman with ductal carcinoma in situ (DCIS) was examined in submacroscopic sections by stereomicroscopic and histologic techniques. Serial 2 mm sections underwent computer-assisted complete three-dimensional reconstruction of all MDLS. RESULTS: The entire breast that was studied contained 16 MDLS that were arranged radially, with the nipple at the center. Of these 16 MDLS, 4 (25.0%) had ductal anastomoses whereas the remaining 12 MDLS had no ductal anastomoses and completely independent regional anatomy. Ductal anastomoses were observed at 11 sites in the 4 MDLS. The 2 of 11 ductal anastomoses that connected different MDLS (18.2%) were situated > 4 cm from the nipple. The remaining nine ductal anastomoses connected ducts within the same MDLS; their location varied from near the nipple to the peripheral region. In the specimen examined, DCIS extended only within a single MDLS and did not spread between different MDLS via ductal anastomoses. CONCLUSIONS: To the authors' knowledge, the current study is the first time the complete architecture of all MDLS in an entire breast has been studied three-dimensionally. The risk of promoting the intraductal spread of disease during surgery may be greater when intraductal lesions extend more peripherally than centrally. The features of ductal anastomoses may provide a significant anatomic clue regarding negative surgical margins in breast conserving surgery. PMID- 11413515 TI - Form or function? Part 1. Subjective cosmetic and functional correlates of quality of life in women treated with breast-conserving surgical procedures and radiotherapy. AB - BACKGROUND: Quality of life (QOL) after a diagnosis of cancer varies considerably across individuals. Treatment-related factors that predict QOL for women who are diagnosed with breast carcinoma require further specification. This study was designed to develop a measure of perceived aesthetic (e.g., breast shape) and functional status (e.g., pain, mobility) after breast-conserving surgical treatment (BCT) and radiotherapy, to examine the relations of these indicators with patient QOL, and to determine whether these relations varied as a function of diagnosis duration. METHODS: Women (n = 185 patients) who underwent BCT and radiotherapy for Stage 0-II disease for whom the diagnosis duration ranged from 3 months to 18 years completed assessments of background information, perceived cosmetic and functional treatment outcomes, and QOL. Medical data also were obtained from medical charts. RESULTS: The Breast Cancer Treatment Outcome Scale (BCTOS) produced a coherent factor structure and three internally consistent subscales (i.e., cosmetic status, functional status, and breast specific pain) that demonstrated predictive validity. With patient age, diagnosis duration, and other BCTOS subscales controlled, greater breast specific pain predicted greater depressive symptoms (P < 0.01) and lower QOL related to mental health (P < 0.05) and physical health (P < 0.05). Cosmetic status predicted QOL related to physical health (P < 0.05). The relations of breast specific pain with QOL indicators varied somewhat as a function of diagnosis duration. CONCLUSIONS: Although considerable research on treatment-relevant outcomes has addressed appearance related concerns, functional parameters have not been explored fully. Findings suggest that functional consequences of treatment, and particularly breast specific pain, also are significant influences on patient QOL. PMID- 11413516 TI - Form or function? Part 2. Objective cosmetic and functional correlates of quality of life in women treated with breast-conserving surgical procedures and radiotherapy. AB - BACKGROUND: Treatment-related factors that influence quality of life (QOL) for women who are diagnosed with breast carcinoma require study. This study was designed to examine the convergent validity of objective and subjective indices of cosmetic and functional status after patients undergo breast-conserving treatment (BCT) and to test the relations of the objective indicators with QOL. METHODS: Women (n = 54 patients) who had received BCT and radiotherapy for Stage 0-II disease for whom the diagnosis duration ranged from 9 months to 18 years completed assessments of background information, perceived cosmetic and functional outcomes, and QOL. They also were administered a clinical breast examination, including objective ratings of arm edema and breast cosmesis. The patients were a subsample from the study reported by the authors in an accompanying article that is presented in this issue. RESULTS: The findings revealed positive cosmetic and functional treatment outcomes, such that 82% of patients had no or minimal arm edema, and 70% of patients evidenced good or excellent cosmetic results. Convergent validity was demonstrated for the objective and subjective assessments of cosmetic and functional treatment outcomes. In addition, women who had more arm edema reported poorer QOL with regard to depressive symptoms (P < 0.05) and fear of disease recurrence (P < 0.05). CONCLUSIONS: The findings from this study and those reported in the accompanying article reveal that functional treatment outcomes, such as arm edema and breast specific pain, are important correlates of QOL even many years after patients undergo BCT and radiotherapy. Both subjective and objective indicators of treatment outcomes are useful predictors of QOL. PMID- 11413517 TI - Phase II trial of epirubicin, cisplatin, oral uracil and tegafur, and leucovorin in patients with advanced gastric carcinoma. AB - BACKGROUND: The results of chemotherapy for patients with gastric carcinoma generally have been modest, although regimens developed more recently have produced higher response rates. One such regimen is epirubicin, cisplatin, and protracted infusion of 5-fluorouracil (ECF). The advantage of a long-term oral administration of uracil and tegafur (UFT) is that this treatment may be used to mimic the protracted infusion of 5-fluorouracil (5-FU). In addition, UFT treatment combined with leucovorin had a favorable activity and tolerable toxicity in patients with advanced gastric carcinoma. Instead of the inconvenience of an infusion pump and intravenous catheter for the protracted infusion of 5-FU, the authors administered UFT plus leucovorin in an ECF regimen for the treatment of patients with advanced gastric carcinoma. METHODS: Fifty-two patients with advanced gastric carcinoma received epirubicin, cisplatin, and oral UFT plus leucovorin. Epirubicin 50 mg/m(2) and cisplatin 60 mg/m(2) were administered on Day 1 by intravenous injection. Tegafur and uracil 360 mg/m(2)/day orally was administered in conjunction with leucovorin administered at a fixed dose of 45 mg/day orally in divided daily doses for 21 days followed by a 7-day rest period. These courses were repeated every 4 weeks. The median age of the patients was 59 years with a median World Health Organization performance status of 1. Patients received a median of five courses of treatment (range, 1 10). RESULTS: Among the 47 patients evaluated, three patients achieved complete response, and 24 patients had partial responses, for an overall response rate of 57.5% (95% confidence interval, 71.5-43.3%). Stable disease was reported in 11 patients (23.4%), and another 9 patients (19.1%) showed disease progression. The median duration of survival was 15 months (range, 2-33+). The main toxicity was nausea/vomiting and neutropenia. Significant toxicity (modified National Cancer Institute common toxicity Grade 3 or 4) included neutropenia in 22 patients (42%), nausea in 14(27%), vomiting in 9 (18%), oral mucositis in 3 (6%), and diarrhea in 3 (6%) patients. CONCLUSIONS: The authors conclude that epirubicin, cisplatin, and oral UFT plus leucovorin, a convenient regimen, has a significant activity and tolerable toxicities in patients with gastric carcinoma. PMID- 11413518 TI - Concurrent hypermethylation of multiple tumor-related genes in gastric carcinoma and adjacent normal tissues. AB - BACKGROUND: Transcriptional silencing by CpG-island hypermethylation now is believed to be an important mechanism of tumorigenesis. To date, studies on CpG island hypermethylation in gastric carcinoma and adjacent normal tissues are few. METHODS: The authors examined 5 gastric carcinoma cell lines, 26 frozen gastric carcinoma tissues and their adjacent nontumor area for concurrent CpG-island hypermethylation in 6 tumor-related genes (p15, p16, E-cadherin, GST-pi, hMLH1, and VHL) by methylation-specific polymerase chain reaction. Nontumorous gastric tissues from 10 gastritis patients were used as controls. RESULTS: Hypermethylation was not detected in any tissue taken from gastritis patients but was identified in all 5 cell lines and in 24 (92.3%) gastric carcinoma patients. CpG-island methylation in tumor-related genes also was detected in 7 out of the 25 adjacent normal tissues from cancer patients. Hypermethylation of E-cadherin, p15, and p16 were detected more frequently than GST-pi and hMLH1, whereas aberrant methylation of VHL was not detected. Concurrent hypermethylation in 2 or more tumor-related genes was detected in 3 out of the 5 gastric carcinoma cell lines, 22 (84.6%) tumor samples, and 5 (20%) adjacent gastric tissues. Eighteen (69.2%) tumor samples showed hypermethylation in >or= 3 genes. CONCLUSIONS: The current study showed that concurrent hypermethylation of multiple tumor-related genes is detected frequently in gastric carcinoma and adjacent normal tissues. Study findings suggested that a mechanism that leads to dysregulation in CpG island methylation is likely to be involved in the early gastric carcinogenesis process. PMID- 11413519 TI - Outcomes for men with clinically nonmetastatic prostate carcinoma managed with radical prostactectomy, external beam radiotherapy, or expectant management: a retrospective analysis. AB - BACKGROUND: With a lack of data from randomized trials, the optimal management of men with nonmetastatic prostate carcinoma is controversial. The authors sought to define the outcomes of three common strategies for managing patients with nonmetastatic prostate carcinoma: expectant management, radiotherapy, and radical prostatectomy. METHODS: The authors conducted a retrospective cohort study with standardized collection of key prognostic data, including centralized assignment of Gleason grades from original biopsy specimens. Participants included all Connecticut hospitals (the expectant management cohort) and three academic medical centers in other states (the radiotherapy and surgery cohorts). Two thousand three hundred eleven consecutive men ages 55-74 years who were diagnosed during 1971-1984 with nonmetastatic prostate carcinoma and were treated at the participating sites were included. RESULTS: Kaplan-Meier estimates with 95% confidence intervals (95% CI) of overall survival at 10 years for each cohort were as follows: expectant management cohort, 42% of patients (95% CI, 38-46%); radiotherapy cohort, 52% of patients (95% CI, 46-58%); and radical prostatectomy cohort, 69% of patients (95% CI, 67-71%); for disease specific mortality, the estimates were as follows: expectant management cohort, 75% of patients (95% CI, 71-79%); radiotherapy cohort, 67% of patients (95% CI, 61-73%); and radical prostatectomy cohort, 86% of patients (95% CI, 84-88%). There were large differences in distributions of important prognostic factors among men in the different treatment groups. CONCLUSIONS: These data provide precise estimates of the outcomes of patients who have been treated with different modalities for nonmetastatic prostate carcinoma in the recent past. Direct comparisons of outcomes between treatment groups are inadvisable because of the different characteristics of patients who select these alternative management strategies. PMID- 11413520 TI - Human papillomavirus as a prognostic factor in carcinoma of the penis: analysis of 82 patients treated with amputation and bilateral lymphadenectomy. AB - BACKGROUND: Many studies have tried to identify prognostic factors to guide the selection of patients at high risk for metastases in penile carcinoma. The authors evaluated human papillomavirus (HPV) status with respect to clinical features, incidence of metastases, and prognosis of penile carcinoma. METHODS: Human papillomavirus DNA was detected using the polymerase chain reaction on paraffin embedded material from 82 patients with penile carcinoma. The following variables were recorded: age, marital status, clinical staging, tumor surgery extension, histologic grade, tumor thickness, lymphatic and venous embolization, corpora cavernosa and corpus spongiosum infiltration, urethral infiltration, mononuclear and eosinophilic infiltrate, and lymph node spread. Follow-up ranged from 0.1 to 453 months. RESULTS: Human papillomavirus DNA was detected in 30.5% (25 of 82) of samples. HPV-16 was the most frequent type detected (13 of 25, 52%). Human papillomavirus DNA positive tumors had less lymphatic embolization by neoplastic cells than HPV negative ones (P = 0.007). The logistic regression revealed that only lymphatic embolization was related to HPV status. There was no difference, however, between HPV DNA negative and HPV DNA positive patients according to presence of lymph node metastases (P = 0.386). No difference was found in the 10-year survival rate (68.4% vs. 69.1%; P = 0.830) between the two groups. CONCLUSIONS: These data suggest that HPV status does not influence prognosis in invasive penile carcinoma. PMID- 11413521 TI - Prediction of prostate carcinoma stage by quantitative biopsy pathology. AB - BACKGROUND: Considerable evidence has shown that the use of computational algorithms to combine pretreatment clinical and pathology results can enhance predictions of patient outcome. The aim of this study was to prove that the application of such methods to predict patient-specific likelihoods of organ confined (OC) prostate carcinoma (PCA) may be helpful to patients and physicians when they are choosing an optimal treatment for carcinoma of the prostate. METHODS: The authors used clinical and quantitative pathology results from the biopsy specimens of 817 PCA patients who had been evaluated at a large national pathology reference laboratory. The pathology parameters that were measured included the number of positive cores, Gleason grades and score, percentage of tumor involvement, and the tumor location. The pathologic stage of these cases, as determined by results from radical prostatectomy, lymphadenectomy, or bone scan, categorized the PCA as either OC, non-OC due to capsular penetration only (NOC-CP) or advanced disease with metastasis (NOC-Mets), i.e., seminal vesicle and/or lymph-node positive or bone-scan positive. There were a total of 481 OC cases, 185 NOC-CP cases, and 151 NOC-Mets cases. Patient-specific prediction models were trained by ordinal logistic regression (OLOGIT) and genetically engineered neural networks (GENNs), and the resulting trained models were validated by biopsy information from an independent set of 116 PCA patients. RESULTS: When the authors applied a cutoff of >or= 35% for the n = 817 training set of OC, NOC-CP, and NOC-Mets predictive probabilities, the OLOGIT model predicted OC PCA with an accuracy of 91%, whereas the GENN model predicted the same with an accuracy of 95%. When the authors employed the n = 116 validation set (76 OCs, 31 NOC-CPs, and 9 NOC-Mets), the OLOGIT and GENN models correctly identified OC PCA with 91% and 97% accuracy, respectively. CONCLUSIONS: The value of combining patient pretreatment diagnostic pathology parameters to make predictions concerning the postoperative extent of pathology was illustrated clearly in this study. This finding further confirms the need to pursue such approaches for PCA disease management in the future, especially with the increasing prevalence of clinical T1c (American Joint Committee on Cancer, 1977) disease. PMID- 11413522 TI - Neoadjuvant chemotherapy for unresectable ovarian carcinoma: a French multicenter study. AB - BACKGROUND: Initial debulking surgery followed by chemotherapy is the current treatment for International Federation of Gynecology and Obstetrics Stage IIIC/IV ovarian carcinoma but has a limited efficacy when optimal cytoreduction is not achieved at the end of the surgical procedure. An alternative treatment for these patients could be neoadjuvant chemotherapy. The purpose of this retrospective study was to report the results of neoadjuvant chemotherapy in operable patients (no medical contraindication to surgery) presenting with primary unresectable tumors. METHODS: Between January 1996 and March 1999, operable patients presenting with Stage IIIC or IV ovarian carcinoma underwent, in six French gynecologic oncology departments, surgical staging to evaluate tumor resectability. When the tumor was deemed unresectable by standard surgery, the patient received three to six cycles of platinum-based neoadjuvant chemotherapy according to the response and the center's usual protocol. Patients were surgically explored after completion of neoadjuvant chemotherapy when the tumor did not progress during treatment. Debulking was performed during this secondary surgery when a response to chemotherapy was observed. RESULTS: Fifty-four patients were treated by neoadjuvant chemotherapy. The first surgical staging procedure was laparoscopy in 33 patients (61%) and laparotomy in 21 patients (39%). The median number of neoadjuvant chemotherapy cycles was 4 (range, 0-6). Forty-three patients (80%) responded to neoadjuvant chemotherapy and then tumors were debulked. Optimal cytoreduction was obtained in 39 patients (91% of the patients who underwent debulking) and with standard surgery in 32 patients (82%). For patients whose tumors were optimally debulked, blood transfusions were administered to 17 patients (43%), median intensive care unit stay was 0 days (range, 0-7 days), and median postoperative hospital stay was 10 days (range, 4 62 days). Median overall survival for the total series was 22 months. Survival was better for patients debulked after neoadjuvant chemotherapy compared with patients with nondebulked tumors (P < 0.001). CONCLUSIONS: Neoadjuvant chemotherapy for primary unresectable ovarian carcinoma leads to the selection of a subset of patients sensitive to chemotherapy in whom optimal cytoreduction can be achieved after chemotherapy by standard surgery in a high proportion of cases. Conversely, aggressive surgery can be avoided in patients with initial chemoresistance, in whom the prognosis is known to be poor regardless of treatment. PMID- 11413523 TI - Completely resected anaplastic thyroid carcinoma combined with adjuvant chemotherapy and irradiation is associated with prolonged survival. AB - BACKGROUND: The prognosis of anaplastic thyroid carcinoma (ATC) has been dismal. The objective of this study was to identify prognostic factors in patients who had prolonged survival. METHODS: Patients with ATC were identified from a computer database at a tertiary referral center. Univariate and multivariate analyses for survival differences were performed using the Kaplan-Meier log-rank statistic and the Cox proportional hazards model, respectively. RESULTS: Of the 33 evaluable patients, median survival was 3.8 months. Median age was 69 years. Prior goiter was present in 6 patients (18%), and 6 (18%) had prior thyroid carcinoma. Median tumor size was 6 cm, and 12 (36%) had adjacent well differentiated carcinoma. Of the 26 patients who underwent neck exploration, 8 patients were potentially cured and received postoperative chemotherapy and irradiation; 4 (50%) were surgically macroscopically free of disease, and 4 (50%) patients had minimal residual disease after total thyroidectomy and resection of tumor adherent to adjacent structures. Four of these 8 patients survived longer than 2 years; their 5-year survival estimate was 50%. Eighteen patients underwent palliative resection of neck disease, leaving macroscopic residual disease or distant metastases; postoperative adjuvant chemotherapy and irradiation were administered in 16 of these 18 patients. Seven patients were treated with only chemotherapy and irradiation. In patients treated with potentially curative resection, median survival was 43 months compared with 3 months with palliative resection (P =0.002); the median survival of 3.3 months with only chemotherapy and irradiation was no different than palliative resection (P =0.63). No association was found between survival and age, prior goiter, prior thyroid carcinoma, adjacent differentiated carcinoma, or tumor size. CONCLUSIONS: Although the prognosis of most patients with ATC continues to be poor, complete resection of ATC combined with postoperative adjuvant chemotherapy and irradiation resulted in long-term survival, even with persistent minimal disease that remained on vital structures. An aggressive attempt at maximal tumor debulking followed by adjuvant therapy was found to be warranted in patients with localized ATC. PMID- 11413524 TI - The significance of granzyme B expression in patients with angiocentric lymphoma of the head and neck. AB - BACKGROUND: The objectives of this study were to establish a correlation between granzyme B expression and the clinicopathologic characteristics of patients with angiocentric lymphomas of the head and neck and to determine whether the expression of granzyme B had any influence on the treatment outcomes of such patients. METHODS: Fifty-seven patients with angiocentric lymphoma of the head and neck who were treated between 1987 and 1996 were divided into two groups according to whether their tumors were immunoreactive for granzyme B: the granzyme B negative group (n = 22 patients) and the granzyme B positive group (n = 35 patients). The clinicopathologic features, immunohistochemical findings, patterns of disease failure, and survival data for the granzyme B positive group were compared with those for the granzyme B negative group. RESULTS: Greater than 60% of patients with angiocentric lymphoma of the head and neck were shown to have granzyme B positive tumors. All tumors that expressed granzyme B also consistently coexpressed CD56, indicating that they probably are the neoplastic equivalent of either natural killer (NK) cells or activated cytotoxic T cells. Although there were no significant differences in histopathologic features or expression of CD45RO and polyclonal CD3-epsilon between the groups, the Epstein Barr virus genomes were detected more frequently in the granzyme B positive group compared with the granzyme B negative group. Despite a similar rate of complete remission after initial treatment, the locoregional recurrence rate of patients in the granzyme B positive group was much higher compared with patients in the granzyme B negative group. In addition, compared with patients in the granzyme B negative group, patients in the granzyme B positive group also had an increased risk of systemic disease recurrence and a decreased overall survival rate. CONCLUSIONS: The data indicate that the cytotoxic granule-associated protein, granzyme B, may be used as an additional marker for identifying NK/T-cell lymphoma and as a prognostic indicator for risk assessment in patients with angiocentric lymphoma of the head and neck. PMID- 11413525 TI - Advanced head and neck carcinoma in women: treatment outcomes may not improve with accelerated hyperfractionated radiotherapy. AB - BACKGROUND: The authors undertook a retrospective study on local tumor control, survival, and complications of conventional irradiation compared with accelerated hyperfractionated irradiation in women with selected head and neck tumor sites. METHODS: One hundred eight consecutive women who were treated with radiation alone for cure during 1974-1998 were analyzed. Patients were excluded who had T1 tumors of the vocal cord and those who were treated with brachytherapy implants. Fifty-nine patients were treated with conventional fractionation once daily (QD) during 1974-1998 with a median dose of 2.1 grays (Gy) per fraction up to a total median dose of 69 Gy in a median overall time of 54 days. Forty-nine patients were treated with accelerated hyperfractionation twice daily (BID) during 1987 1998 at a median dose of 1.6 Gy per fraction BID, with an interfraction interval of 4-6 hours, for a total median dose of 66 Gy in 35 days. Patients were not randomized into the QD group or the BID group. RESULTS: The 7-year actuarial local control (LC) rates for T1-T2 tumors in QD-treated and BID-treated patients were 79% and 87%, respectively (P = not significant [NS]). For T3-T4 tumors, the LC rates at 7 years were 59% and 56% for the QD and BID groups, respectively (P = NS). A Cox regression analysis for LC showed that the significant variables were T classification and overall time. Schedule (QD or BID), total dose, dose per fraction, and patient age were not significant variables. For the QD and BID groups, the 7-year actuarial cause specific survival rates for patients with Stage I-II disease were 100% and 65%, respectively (P = 0.004), and, for patients with Stages III-IVA,IVB disease, the rates were 39% and 56%, respectively (P = NS), respectively. Acute morbidity was higher with the BID schedule: In the BID group, 8% of patients required tube or parenteral feeding, and 0% of patients in the QD group required such feeding (P = 0.04). The 5-year actuarial probability of Grade 3-5 late effects (according to the Radiation Therapy Oncology Group/European Organization for Research and Treatment of Cancer grading system) was 4% for the BID group and 0% for the QD group (P = NS). CONCLUSIONS: This study suggests that accelerated hyperfractionated irradiation for women with advanced carcinoma of the head and neck does not provide significantly better local tumor control or cause specific disease free survival compared with conventional fractionation. Women with these malignancies appear to have a better prognosis compared with men. PMID- 11413526 TI - Short-term effects of radiotherapy on attention and memory performances in patients with brain tumors. AB - BACKGROUND: The objective of the current study was to evaluate the short-term effects of radiotherapy on attention and memory performance in patients with brain tumors. METHODS: Thirteen patients underwent a neuropsychologic examination on three occasions: at baseline before radiotherapy, which lasted for 6 weeks; within 2 weeks after the completion of radiotherapy; and 3 months after the completion of radiotherapy. Thirty healthy individuals were studied at baseline as a control group. Verbal, visuomotor, and memory tests were administered. Attentional performances were examined using CogniSpeed reaction time software. RESULTS: At baseline, the patients scored significantly lower than the controls in tests of semantic memory (Similarities; P = 0.002), visuoconstructive skill (Block Design; P = 0.028), and visual and verbal memory (recall of 20 objects: immediate, P = 0.017; delayed, P = 0.002; recall of 30 paired word associates: immediate, P = 0.002; delayed, P = 0.005). The patients performed significantly more poorly than the control group on the Two-Choice Reaction Time test (P = 0.047), the Subtraction test (P = 0.042), and the Vigilance test (P = 0.023). After radiotherapy, the performance of the patients did not worsen on any measure, whereas their conscious cognitive processing was improved (P = 0.002). CONCLUSIONS: Treatment with radiotherapy did not cause negative short-term effects on attentional and memory functions in patients with brain tumors who received a radiation dose of 54.5 grays. The cognitive deficits found in these patients were evident at baseline before radiotherapy. Thus, it seems likely that the deficits were associated with the tumor itself and/or the surgical procedures. PMID- 11413527 TI - Pharmacokinetic study of intralesional cisplatin for the treatment of hepatocellular carcinoma. AB - BACKGROUND: In the current study the authors examined the pharmacokinetics of direct intralesional injection of cisplatin/epinephrine/bovine collagen gel in patients with hepatocellular carcinoma and cirrhosis. METHODS: Six patients with cirrhosis and unresectable hepatocellular carcinoma received a direct intralesional injection (range, 6.7-26.7 mg) into their tumors under ultrasonographic guidance. The authors determined the total cisplatin (Pt) concentration in the plasma and urine and nonprotein-bound free Pt in plasma ultrafiltrate using flameless atomic absorption spectrometry. Data from individual patients were analyzed to calculate the pharmacokinetic parameters via a noncompartmental method for constant infusion. To demonstrate that the changes in pharmacokinetics are not related to the underlying cirrhosis, a similar methodology was applied to measure the pharmacokinetic parameters of four similar patients who were treated with cisplatin, 75 mg/m(2), as a 1-hour intravenous infusion. RESULTS: The time to attain maximum concentration of total Pt after intralesional injection was dose-dependent and ranged from 2-13 hours. The concentration-time curve was biphasic in nature. The initial half-life of total Pt in patients who received an intralesional injection varied with the cisplatin dose. The initial half-life for cisplatin doses < 15 mg was approximately 9 hours and the initial half-life at higher cisplatin doses (> 15 mg) was approximately 25 hours. The area under the curve (AUC) was dose-dependent with values ranging from 38-150 microm/mL x hour. Pharmacokinetic parameters for free Pt (ultrafiltrate) were significantly different. The time to attain maximum concentration (t-max) and terminal half-life were shorter and the average AUC was approximately 100-fold lower than total Pt. After the intravenous infusion of cisplatin, the t-max for total and free Pt was 1.3 hours and 1.1 hours, respectively. The terminal half-life and average AUC for total Pt was 194 hours and 247 microg/mL per hour, respectively, and its corresponding parameters for free Pt after intravenous infusion were much lower, similar to the findings for the intralesional injection. CONCLUSIONS: The prolonged t-max and initial half life noted with the intralesional injection of cisplatin/epinephrine/collagen gel are consistent with its proclaimed ability to retain cisplatin at the tumor and delay its release in systemic circulation. The kinetics of intralesional cisplatin injection also suggest local sequestration of the drug in the injected site. Parameters of intravenous cisplatin infusion in cirrhotic patients are similar to those of patients from the historic control group. PMID- 11413528 TI - Adjuvant chemotherapy after resection of hepatocellular carcinoma causes deterioration of long-term prognosis in cirrhotic patients: metaanalysis of three randomized controlled trials. AB - BACKGROUND: The goal of the current study was use metaanalysis to evaluate the effect of postoperative adjuvant chemotherapy after resection of hepatocellular carcinoma (HCC), especially in cirrhotic patients. METHODS: One hundred-eight patients with radical resection of HCC were included in 1 of 3 prospective randomized control studies that used different postoperative chemotherapy protocols. Fifty-one patients underwent resection alone, and 57 patients received postoperative adjuvant chemotherapy. The first protocol was intraarterial epirubicin (40 mg/m(2)) at 1 month after surgery followed by oral tegafur (300 mg/day) for 1 year. The second protocol was intraarterial epirubicin (40 mg/m(2)) at 1 month after resection followed by intravenous epirubicin (40 mg/m(2)) every 3 months for 2 years. Additionally, carmofur 300 mg/day was administered for 2 years. The third protocol was intravenous epirubicin (40 mg/m(2)) every 2 months starting 1 month after surgery for 1 year. RESULTS: There were no significant differences in clinicopathologic background in each of the three protocols between the groups with and without chemotherapy. Postoperative chemotherapy using the current protocols failed to improve patient outcome in all patients and failed to improve disease-free and overall survival in any patients who were included in individual protocols. In patients with cirrhosis, postoperative chemotherapy was associated with significantly worse disease-free (P = 0.0376) and overall survival rates (P = 0.0077). CONCLUSIONS: The current study indicated that cancer recurrence in the remnant liver is enhanced and the long-term outcome is deteriorated by postoperative chemotherapy after resection of HCC in cirrhotic patients. PMID- 11413529 TI - Association between insulin-like growth factor-2 and metastases after transcatheter arterial chemoembolization in patients with hepatocellular carcinoma: a prospective study. AB - BACKGROUND: Hypoxia up-regulates insulin-like growth factor-2 (IGF-2) and thus stimulates the growth of hepatocellular carcinoma (HCC) cells. In the current study, the authors prospectively evaluated changes in plasma IGF-2 levels in HCC patients after transcatheter arterial chemoembolization (TACE), which usually results in hypoxic insult to liver tissue. The authors also examined the association between changes in plasma IGF-2 levels after TACE and HCC progression, especially in relation to metastasis. METHODS: Plasma IGF-2 levels were measured before and 4 weeks after TACE in 46 patients with HCC. Three months after TACE, the patients were evaluated for the occurrence of metastatic HCC. RESULTS: In 13 of the 46 patients, post-TACE plasma IGF-2 levels decreased by > 20% (Group 1) compared with their basal levels; in 18 patients, the IGF-2 levels changed within 20% (Group 2) and in 15 patients the IGF-2 levels increased by > 20% (Group 3). Plasma IGF-2 levels had a tendency to increase in patients with large-sized tumors, high serum alpha-fetoprotein (AFP) levels, and the heterogeneous uptake of iodized oil. Metastatic foci were found in 9 patients in Group 3 (60%), in contrast to only 3 patients in Group 2 (17%) and in none of the patients in Group 1 (P = 0.001). On multivariate analysis, higher Child-Pugh scores and increased plasma IGF-2 levels (Group 3) were found to be independent risk factors for metastasis. CONCLUSIONS: Increased plasma IGF-2 levels after TACE, which are common in patients with large-sized tumors and high serum AFP levels, appear to be associated with the occurrence of metastatic HCC after TACE. PMID- 11413530 TI - Patterns of relapse of N2 nonsmall-cell lung carcinoma patients treated with preoperative chemotherapy: should prophylactic cranial irradiation be reconsidered? AB - BACKGROUND: Although it induces a relevant reduction in the risk of both visceral metastases and locoregional recurrences, the combination of chemotherapy and surgery only marginally improves the survival of patients with Stage IIIA(N2) (International Union Against Cancer staging and classification system) nonsmall cell lung carcinoma (NSCLC). The purpose of the current study was to analyze the patterns of relapse in these patients. METHODS: In this study, the authors compared the patterns of relapse in 81 patients with clinically detectable N2 NSCLC who had been treated with preoperative chemotherapy with the relapse patterns of 186 comparable patients who had been treated with primary surgery. Clinically detectable N2 (cN2) denotes mediastinal lymph node enlargement on computed tomography scan, which then is confirmed by mediastinoscopy. RESULTS: Overall 20% of patients developed a locoregional recurrence. Chemotherapy decreased the risk of visceral metastasis as 28% of the patients preoperatively treated and 38%of those not treated with preoperative chemotherapy presented a visceral metastasis (P < 0.05). Preoperative chemotherapy and adenocarcinoma subtypes were associated with a higher rate of brain metastasis (P < 0.05). Thirty-two percent of the patients treated preoperatively and 18% of those not treated with preoperative chemotherapy presented a brain metastasis (P < 0.05), which was isolated in 22% and 11% of the patients, respectively (P < 0.05). CONCLUSION: The current study found that preoperative chemotherapy for cN2 decreases the risk of visceral metastasis but is associated with a high rate of isolated brain metastases. Prophylactic cranial irradiation may need to be reinvestigated in clinical trials, especially in patients who present with an adenocarcinoma. PMID- 11413531 TI - The sentinel lymph node status is an important factor for predicting clinical outcome in patients with Stage I or II cutaneous melanoma. AB - BACKGROUND: In a cohort of patients, the authors investigated whether and to what extent the sentinel lymph node (SLN) status contributes to predicting the probability of remaining disease free for at least 3 years. In addition, several traditional prognostic factors were analyzed: Breslow thickness, Clark invasion level, ulceration, lymphatic invasion, location, type of the melanoma, and age and gender of the patient. METHODS: In 263 consecutive patients with proven American Joint Committee on Cancer Stages I and II cutaneous melanoma, the triple technique SLN procedure was used, i.e., preoperative visualization of the lymph channels from the initial site of the melanoma toward the SLN by (dynamic) lymphoscintigraphy, intraoperative visualization of those particular lymph channels and lymph nodes with blue dye, and a gamma probe to measure accumulated radioactivity in radiolabeled lymph nodes. Median follow-up time was 48 months (range, 36-84 months). Multivariate logistic regression analysis was performed to examine the influence of the SLN status and several other prognostic factors on a minimum 3-year disease free survival. RESULTS: In 20% of patients, the SLN proved to be tumor positive. For SLN negative patients, the 5-year disease free survival rate was 91% (+/- 2.4%), and for SLN positive patients it was 49% (+/- 9%). Five variables showed a strong and statistically significant independent prognostic association with outcome, i.e., SLN status (P = 0.0007), thickness of primary melanoma (1.01-2.0 mm; P = 0.04), ulceration (P = 0.05), and lymphatic invasion (P = 0.01) of primary melanoma, and age (40-50 years; P = 0.01). CONCLUSIONS: The SLN status-along with Breslow thickness, ulceration, lymphatic invasion, and age- seems to have strong additional value in predicting a minimum 3-year disease free period after the SLN procedure. Patients with a positive SLN have a poorer prognosis than those with a negative SLN. PMID- 11413532 TI - Efficacy of ultrasound B-scan compared with physical examination in follow-up of melanoma patients. AB - BACKGROUND: The value of ultrasound B-scan for routine follow-up of melanoma patients still is not generally accepted. Therefore, the authors compared the efficacy of physical examination (PE) with ultrasound B-scan (UBS) for detection of regional tumor recurrence in melanoma patients. The aim of the current study was to evaluate whether early detection of metastases improves relapse-free and overall survival. METHODS: For a period of 4 years, 829 consecutive melanoma patients were followed prospectively. Physical examination of 3011 patients and concomitant UBS of in-transit routes and regional lymph node basins were performed. Suspicious lesions were diagnosed by fine-needle aspiration cytology and pathology. RESULTS: During the study period, 242 (90.6%) of 267 patients with melanoma recurrences were first recognized by PE or UBS within the routine follow up program. The sensitivities of both methods differed significantly (P = 0.001). Metastases were detected by PE in only 61 of 242 recurrences (25.2%, 95% confidence interval [CI]: 19.9-31.2%), whereas UBS revealed 240 recurrences (99.2%, 95% CI: 97.3-99.6%). The specificity was 98.4% (95% CI: 97.8-98.8%) and 98.3% (95% CI: 97.7-98.7%), respectively. Survival of 103 patients who presented with a first nodal melanoma recurrence was followed and analyzed by multiple Cox regression. Overall survival was affected by the diameter of the largest metastasis (P = 0.001) and the number of metastatic lesions (P = 0.012). CONCLUSION: The study found that ultrasound B-scan was highly effective in the early detection of regional melanoma metastases compared with physical examination. Earlier detection of such metastases seemed to result in improved overall survival. PMID- 11413533 TI - Tumor-infiltrating lymphocytes are a marker for microsatellite instability in colorectal carcinoma. AB - BACKGROUND: Cells with deficient DNA mismatch repair develop microsatellite instability. Extensive microsatellite instability (MSI-high) is characteristic of colorectal carcinomas in hereditary nonpolyposis colorectal carcinoma (HNPCC) and in 10-% 15% of sporadic colorectal carcinomas. Microsatellite instability-high colorectal carcinomas differ from others in important clinical and pathologic features. However, MSI typing is expensive and not widely available. Microsatellite instability type may be predicted by tumor-infiltrating lymphocytes (TILs), which can be evaluated with ordinary light microscopy. METHODS: The authors evaluated TILs as a pathology screen for MSI-high status in 138 colorectal carcinomas that had been evaluated for MSI in a variety of studies. This case series was systematically enriched with HNPCC and other MSI high cases to allow accurate sensitivity and specificity estimation. Tumor infiltrating lymphocytes were quantitated as TILs per 10 high-power microscopic fields by an observer blinded to MSI status. RESULTS: Of the 138 carcinomas studied, 67 (48.6%) were MSI-high, 22 (15.9%) were MSI-low, and 49 (35.5%) were MSI-stable. All 25 HNPCC colorectal carcinomas were MSI-high. Tumor-infiltrating lymphocytes counts ranged from 0 to 300, with a markedly skewed distribution (median, 11; mean, 36). Sensitivity and specificity for selected cut points of TIL count were computed. Using a TIL count of 5 as a cut point yields a sensitivity of 93% and specificity of 62%. In a population in which 12% were MSI high, consideration of TIL could reduce the number of colorectal carcinomas referred for MSI testing by greater than one-half, and still 93% of the MSI-high carcinomas would be identified. CONCLUSIONS: The presence of MSI defines a subset of colorectal carcinomas with special molecular etiology and characteristic clinical, pathologic features, inclusive of increased survival. The authors conclude that quantification of TILs may provide a simple, single criterion for choosing which colorectal carcinomas are candidates for MSI testing. PMID- 11413534 TI - The role of surgery and postoperative chemoradiation therapy in patients with lymph node positive esophageal carcinoma. AB - BACKGROUND: Patients who have undergone resection for lymph node positive esophageal carcinoma are at high risk of disease recurrence and early death. The role of postoperative adjuvant therapy in this population needs to be determined. METHODS: A retrospective review of all patients with resected esophageal carcinoma between 1991 and 1997 was performed. Lymph node positive (N1) patients who received concurrent or sequential postoperative radiotherapy (50 grays) and chemotherapy (cisplatin, 5-fluorouracil with or without epirubicin) were compared with N1 patients who underwent surgery alone. The disease free and overall survival rates were calculated using the Kaplan-Meier method, and groups were compared with the log-rank test. Prognostic variables were entered into a Cox regression model controlling for age, weight loss, T status, Eastern Cooperative Oncology Group (ECOG) score, and treatment received. RESULTS: A total of 165 patients were reviewed: Twenty-eight N1 patients underwent surgery alone (S group), and 38 N1 patients underwent surgery and received postoperative chemoradiation therapy (CRT group). Preoperative risk factors, tumor characteristics, ECOG scores, and lengths of hospital stay were similar. The disease free survival rates were similar (S group, 10.6 months; CRT group, 10.2 months), although the S group had more local disease recurrences (S group, 35%; CRT group, 13%; P = 0.09). The overall survival rate according to the Kaplan Meier analysis showed a significant survival advantage with postoperative CRT radiation (log-rank test; P = 0.001). The median overall survival for the CRT group was 47.5 months, which was significantly longer than that of the S group (14.1 months). The ECOG score, T status, and treatment received all were found to influence survival significantly on univariate analysis. In the multivariate model, postoperative CRT was a predictor of survival (P = 0.007; risk ratio for mortality, 0.35; 95% confidence interval, 0.16-0.76) and was correlated with a significantly decreased risk of death in patients with lymph node positive, resected esophageal carcinoma. CONCLUSIONS: Postoperative CRT appears to prolong survival in patients with lymph node positive, resected esophageal carcinoma. PMID- 11413535 TI - A Phase I trial of pulse calcitriol in patients with refractory malignancies: pulse dosing permits substantial dose escalation. AB - BACKGROUND: Calcitriol is the principal biologically active metabolite of vitamin D. Calcitriol's activity against many neoplasms is well documented, but calcitriol's therapeutic application has been hampered by predictable hypercalcemia when it is given daily. Because laboratory data has suggested that intermittent exposure to high levels of calcitriol may be sufficient to produce antiproliferative effects, the authors developed a Phase I trial to determine the maximal tolerated dose, dose-limiting toxicity, and the pharmacokinetic profile of calcitriol given weekly by mouth. METHODS: Patients with refractory malignancies were enrolled for 4 weeks of treatment followed by 4 weeks of observation. Reenrollment at a higher dose level was permitted for patients who had evidence of response or stable disease and no Grade 3 or greater toxicity. The starting dose was 0.06 microg/kg. RESULTS: Fifteen patients received 20 cycles of therapy. Doses up to 2.8 microg/kg of calcitriol weekly produced no dose-limiting toxicity. While peak levels and the area under the serum concentration-time curve of calcitriol increased in a linear fashion at lower doses, saturable absorption was observed at doses above 0.48 microg/kg. Doses of 0.48 microg/kg and above produced mean peak calcitriol levels of 1625 pg/mL, approximately 25-fold greater than top normal levels and well within the therapeutic range suggested by in vitro experiments. Eight patients experienced self-limiting Grade 1 hypercalcemia. CONCLUSIONS: Weekly dosing of oral calcitriol permitted substantial dose escalation with minimal toxicity. Peak serum calcitriol levels were in the predicted therapeutic range. A dose of 0.5 microg/kg was selected for evaluation in Phase II studies. PMID- 11413536 TI - Bulky disease is an adverse prognostic factor in patients treated with chemotherapy comprised of cyclophosphamide, doxorubicin, vincristine, and prednisone with or without radiotherapy for aggressive lymphoma. AB - BACKGROUND: In the current study, the authors analyzed prognostic factors in patients with aggressive lymphoma treated with a chemotherapy regimen comprised of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) with or without radiotherapy. METHODS: Between September 1988 and December 1996, 294 patients with newly diagnosed, clinical Ann Arbor Stage I-IV, aggressive lymphoma were enrolled on 2 protocols at The University of Texas M. D. Anderson Cancer Center. Patients on these studies had a relatively favorable prognosis; 100% had M. D. Anderson tumor scores 1 extranodal site of disease (P < 0.001), bulky disease (>or= 7 cm) (P = 0.005), and an at least 10% elevation in the serum lactate dehydrogenase (LDH) level (P = 0.007). Patient age > 60 years (P = 0.001), bulky disease (P = 0.016), and an at least 10% elevation in the serum LDH level (P = 0.040) also were found to be independent prognostic factors for overall survival. CONCLUSIONS: The independent prognostic factors in the current study suggest that either the M. D. Anderson tumor score system or the IPI can be used to select which aggressive lymphoma patients are at high risk for disease recurrence based on their having more than two adverse factors and who consequently are candidates for more intensive frontline therapy. Involved-field radiotherapy should be considered in those patients with bulky lymphomas. PMID- 11413537 TI - Effect of myocutaneous inflammatory changes caused by intra-arterial chemotherapy on the outcome of patients who undergo limb-saving surgery. AB - BACKGROUND: This study examined the effect of myocutaneous inflammatory changes caused by intra-arterial chemotherapy on the outcome of patients who undergo limb saving surgery. METHODS: One hundred seven patients with malignant bone and soft tissue tumors were administered intra-arterial cisplatin and caffeine preoperatively with or without doxorubicin. Nine patients (8.4%) who had had myocutaneous inflammatory reactions were reviewed to determine the effects of this complication on limb-saving surgery. RESULTS: The patients complained of unbearable and continuous pain while undergoing intra-arterial infusion. Gait disturbances and muscle weakness were caused by their severe regional pain, resulting in the use of crutches even before surgery. Extended areas of necrotic skin and muscle were resected simultaneously with limb-saving surgery on four patients. Myocutaneous necrosis in one patient, indurations in two patients, and flares in two patients were not treated surgically. Pain relief was provided to eight patients at some point. Four patients with extensive myocutaneous necrosis around the knee joint had restricted range of motion and poor functional results. Radionuclide angiography using 99m-Technetium-macroaggregated albumins was performed to evaluate the blood flow to the affected muscle and tumor. It showed marked increase of the radioisotope perfusion in the affected muscles but little uptake in the tumor. These results may suggest that the affected muscles diminish the effects of anticancer drugs on the tumors. CONCLUSIONS: Myocutaneous inflammatory reactions should be prevented if possible to obtain good limb function and chemotherapeutic effects in patients who undergo intra-arterial chemotherapy. Resection of necrotic tissue is mandatory to relieve pain. PMID- 11413538 TI - What constitutes optimal therapy for patients with rhabdomyosarcoma of the female genital tract? AB - BACKGROUND: Factors affecting outcome for rhabdomyosarcoma (RMS) of the female genital tract in patients treated on Intergroup Rhabdomyosarcoma Study Group (IRSG) protocols I-IV were evaluated to define optimal therapy. METHODS: Records of 151 patients with tumors of the female genital tract who were treated on IRSG protocols I-IV were reviewed for details regarding chemotherapy, surgery, radiotherapy (RT), and outcome. RESULTS: The overall 5-year survival was 82%, (87% for patients with locoregional tumors). Chemotherapy was primarily vincristine, actinomycin-D, and cyclophosphamide (VAC) based. Local therapy was surgery alone in 42% of patients, surgery plus RT in 19% of patients, biopsy plus RT in 12% of patients, and biopsy without RT in 21% of patients. The rate of hysterectomy decreased from 48% in IRS-I/II to 22% in IRS-III/IV with an increase in the use of RT from 23% in IRS-II to 45% in IRS-IV and continued excellent survival. Many patients with vaginal primary tumors received delayed RT or had it omitted on later studies with excellent outcome. For patients with localized embryonal/botryoid tumors, there were no significant differences in 5-year survival among patients with tumors at different sites or among patients treated on IRS-I-IV. In patients with Group I-III tumors, 43% of deaths were from toxicity. Analysis of prognostic factors, with toxic deaths censored, revealed that an age of 1-9 years at the time of diagnosis, noninvasive tumors, and the use of IRS-II or IRS-IV treatments were associated significantly with better outcome. Patients ages 1-9 years fared best (5-year survival of 98%) and patients outside of this age range especially benefited from the intensified therapy used in IRS-III or IRS-IV (5-year survival of 67% on the IRS-I/II vs. 90% in IRS III/IV). CONCLUSIONS: Localized female genital RMS usually is curable with combination chemotherapy, a conservative surgical approach, and the use of RT for selected patients. PMID- 11413539 TI - Survival patterns of cancer patients. AB - BACKGROUND: Optimal means of modeling death rates of large populations with a specific disease have not been described in the literature. METHODS: Statistical modeling was used on archival data. RESULTS: In the authors' prior publications describing the survival of untreated cancer patients, data that were adequately fitted by an exponential curve were found to be much better fitted by an inverse Gompertz curve (R(2) = 99.7% for untreated breast carcinoma, 99.9% for untreated cervical carcinoma). However, when data from treated patients are examined, fits show that successive stages begin at successive positions on the inverse Gompertz curve. Breast carcinoma data showed that treatment begun at an early stage raised survival to a linear decline; at an intermediate stage led to a modified inverse Gompertz, the earlier the stage at which therapy was begun, the greater the survival rate; and at a late stage exhibited an exponential decline showing a negligible effect of late treatment. Confidence in this approach was enhanced by its applicability using published data from the National Cancer Database for breast, pancreatic, bone, and skin cancers. CONCLUSIONS: The use of data modeling allowed us to assess realistically the value of intervention in cancer populations and to optimize staging schemas. It strongly reinforced the concept that early detection provides a far greater impact on a population's subsequent survival than therapy of advanced disease. PMID- 11413540 TI - Lung carcinoma in African Americans: a review of the current literature. PMID- 11413541 TI - Calcium-dependent neurotransmitter release: synaptotagmin to the rescue. PMID- 11413542 TI - Mutations in the second C2 domain of synaptotagmin disrupt synaptic transmission at Drosophila neuromuscular junctions. AB - The synaptic vesicle protein, synaptotagmin, has been hypothesized to mediate several functions in neurotransmitter release including calcium sensing, vesicle recycling, and synaptic vesicle docking. These hypotheses are based on evidence from in vitro binding assays, peptide and antibody injection experiments, and genetic knockout studies. Synaptotagmin contains two domains that are homologous to the calcium ion (Ca(2+))-binding C2 domain of protein kinase C. The two C2 domains of synaptotagmin have broadly differing ligand-binding properties. We have focused on the second C2 domain (C2B) of synaptotagmin I, in particular, on a series of conserved lysine residues on beta-strand 4 of C2B. This polylysine motif binds clathrin-adapter protein AP-2, neuronal calcium channels, and inositol high polyphosphates. It also mediates Ca(2+)-dependent oligomerization. To investigate the importance of these lysine residues in synaptic transmission, we have introduced synaptotagmin I (syt) transgenes harboring specific polylysine motif mutations into flies otherwise lacking the synaptotagmin I protein (syt(null)). Electrophysiological analyses of these mutants revealed that evoked transmitter release is decreased by approximately 36% and that spontaneous release is increased approximately twofold relative to syt(null) flies that express a wild type syt transgene. Synaptotagmin expression in both the mutant and the wild type transgenic lines was equivalent, as measured by semiquantitative Western blot analysis. Thus, the alteration in synaptic transmission was due to the mutation and not to the level of synaptotagmin expression. We conclude that synaptotagmin interactions mediated by the C2 B polylysine motif are required to attain full synaptotagmin function in vivo. PMID- 11413543 TI - Postnatal growth of intrinsic connections in mouse barrel cortex. AB - Surprisingly little is known about the development of connections within a functional area of the cerebral cortex. We examined the postnatal growth of connections in mouse barrel cortex during the second and third weeks after birth, coinciding with the period of rapid synaptogenesis that occurs just after the barrels first form. A barrel is a group of neurons in layer 4 of somatosensory cortex that is part of a cortical column. Each whisker/barrel column is linked anatomically and functionally to a homotopic whisker on the contralateral face. Radial groups of cortical neurons were labeled with the neuronal tracer biotinylated dextran amine in mice ranging in age from postnatal day 8 (P8; P0 is the date of birth) to adulthood. The spatial distributions of retrogradely labeled neurons in different laminae were analyzed. The barrel map in layer 4 was used as a template to compare quantitative data from different animals and to account for substantial changes in barrel and barrel field size during development. Intrinsic projections 1) innervate increasingly more distant targets within barrel cortex up to 3 weeks of age; 2) continue to form in targets after 3 weeks, effectively strengthening existing connections; 3) follow a timetable for growth that is layer-specific; 4) link more distant barrel columns in layer 4 from neurons that are found preferentially in the barrel side and the septa between barrels; and 5) form over the shortest distances between the barrel columns. These data indicate that intrinsic connections in mouse barrel cortex develop by the progressive addition of neuronal connections rather than by sculpting preliminary connections. We describe statistically significant changes in connectivity during development that may be applied to model and assess the development of connections after a variety of experimental perturbations, such as to the environment and/or the genome. PMID- 11413544 TI - Cell death in the development of the posterior cortex in male and female rats. AB - Work from our laboratory has shown that adult male rats have 19% more neurons than female rats in the binocular region and 18% more in the monocular region of the primary visual cortex (Reid and Juraska [1992] J Comp Neurol 321:448-455; Nunez et al., [1999] Soc Neurosci Abstr 25:229). In the current experiment, we investigated whether cell death in male and female rats (postnatal days 2-35) contributes to the formation of these differences. Using stereological techniques, we investigated neuron density along with pyknotic and apoptotic (TdT mediated deoxyuridine triphosphate nick end-labeled) cell density in the developing posterior cortex (future primary visual cortex). Although no sex differences in neuronal density were found in early development, we observed a differential time course of cell death between the sexes. Consistent with earlier reports, males displayed a rapid rise in cell death, with a peak on day 7 followed by a sharp decline to negligible levels by day 15. Females, however, displayed moderate peaks of cell death on days 7 and 11, with the persistence of low-to-modest levels until day 25. Similar patterns were obtained from both pyknotic and apoptotic cell quantification. Also, a formula was developed to estimate the percentage of cells that die during development and the amount of time a dying cell is visible. This study demonstrates that there is a prolonged period of cell death in the posterior cortex of developing female rats that appears to result in more cell death in females than males. This may be an important mechanism by which the sex difference in adult neuron number is created. PMID- 11413545 TI - Abnormal dispersion of a purkinje cell subset in the mouse mutant cerebellar deficient folia (cdf). AB - Purkinje cells of different molecular phenotypes subdivide the cortex of the cerebellum both rostrocaudally into parasagittal bands and mediolaterally into transverse zones. Superimposed on the Purkinje cell compartmentation, the cerebellar cortex is pleated into a reproducible array of lobes and lobules. During cerebellar development, Purkinje cell bands are formed through the rostrocaudal dispersal of embryonic clusters, triggered primarily by a Reelin dependent signaling pathway. In the naturally occurring mouse mutant cerebellar deficient folia (cdf), there is a failure of Purkinje cell dispersion that results in widespread Purkinje cell ectopia in the adult. The ectopia is restricted primarily to that subset of Purkinje cells that does not express zebrin II/aldolase C and that forms ectopic clusters in among the cerebellar nuclei. Most Purkinje cells that express zebrin II are located normally in a monolayer. Thus, the cerebellum of cdf mutants has a failure of Purkinje cell dispersion that is confined primarily to a zebrin II-negative (zebrin II(-)) subpopulation. Despite the Purkinje cell ectopia, the parasagittal band organization of the cerebellum is still clear. The shortening of the cortex is distributed evenly over all lobules, with the result that transverse expression boundaries are relocated with respect to the lobules and fissures. The number of Purkinje cells in the cdf/cdf cerebellum is similar to the number in littermate controls. Therefore, it appears that the lesion in cdf results in the failure of a zebrin II(-) Purkinje cell subset to disperse either due to a cell intrinsic defect or due to an abnormal interaction between the Purkinje cells and either granule cells or afferent inputs. PMID- 11413546 TI - Zona incerta: Substrate for contralateral interconnectivity in the thalamus of rats. AB - We have shown previously that the zona incerta (ZI), a small nucleus deriving from the ventral thalamus, has extensive ipsilateral connections with the higher order and intralaminar nuclei of the dorsal thalamus and that there are many ipsilateral interconnections between the different cytoarchitectonic sectors of the ZI. In this study, we explore the contralateral connections that the ZI has with its opposing nucleus as well as with the other nuclei of the thalamus. Injections of biotinylated dextran or cholera toxin subunit B were made into each of the different ZI sectors (rostral, dorsal, ventral, and caudal) and into intralaminar and higher order dorsal thalamic nuclei of Sprague-Dawley rats by using stereotaxic coordinates. Brains were fixed in aldehyde and processed using standard methods. Our results show that, after injections limited to a given ZI sector, labelled terminal-like elements and cells were seen across the other sectors of the ZI of the contralateral side. Furthermore, after each of these ZI injections, labelling was seen in the intralaminar (e.g., parafascicular, central lateral, and central medial) and higher order (e.g., posterior thalamic, lateral posterior, and lateral dorsal) nuclei of the contralateral side. These patterns of labelling were confirmed after tracer injections into intralaminar and higher order nuclei; after such injections, labelling was seen in the contralateral ZI. In all cases, there was labelling on the ipsilateral side as well, and this was generally heavier than on the contralateral side. Overall, our results indicate that there is a network of interconnections between the ZI of both sides of the thalamus and that the ZI has contralateral connections with the intralaminar and higher order nuclei. Hence, the ZI furnishes a substrate that spreads activity to both sides of the brain. PMID- 11413547 TI - Distribution of estrogen receptor beta immunoreactivity in the rat central nervous system. AB - The discovery of estrogen receptor beta (ER beta) and subsequent localization of its mRNA in the rat central nervous system (CNS) has provided new insights about estrogen action in brain. A critical step in understanding the role of ER beta is demonstrating that the mRNA is translated into functional protein. The present study used a new ER beta-specific polyclonal antiserum (Z8P) and immunocytochemistry (ICC) to investigate the distribution of ER beta in the rat CNS. Ovariectomized female rats were perfusion fixed, and free-floating sections were incubated with Z8P. After visualization with a standard ABC method, nuclear immunoreactivity was seen in neurons throughout the brain, including the olfactory nuclei, laminae IV-VI of the cerebral cortex, medial septum, preoptic area, bed nucleus of the stria terminalis, supraoptic nucleus, paraventricular nucleus, zona incerta, medial and cortical amygdaloid nuclei, cerebellum, nucleus of the solitary tract, ventral tegmental area, and spinal trigeminal nucleus. Moreover, the results of a double-label ICC/ in situ hybridization study revealed that ER beta mRNA and immunoreactivity were colocalized in neurons of the brain, thus confirming the specificity of the antiserum. Through the use of Western blot analysis, Z8P was shown to recognize in vitro translated ER beta, but not ER alpha, as well as a 60-kDa protein from rat granulosa cells and ovary extracts. The results of these studies have demonstrated that (1) ER beta mRNA is translated into immunoreactive protein throughout the rat brain, and (2) ER beta resides in the cell nucleus. Together, these data provide an anatomic foundation for future studies and advance our understanding of estrogen action in hypothalamic and extrahypothalamic brain regions. PMID- 11413548 TI - Elimination of Bax expression in mice increases cerebellar purkinje cell numbers but not the number of granule cells. AB - Cerebellar Purkinje cells and granule cells have been studied extensively as models for investigating neuron-target interactions and the regulation of cell numbers in the developing central nervous system. Recent studies of transgenic mice that overexpress a human Bcl-2 transgene in Purkinje cells suggest both that programmed cell death plays an unexpected role in regulating Purkinje cell number and that Purkinje cells influence the number of granule cells. The role of cell death-related proteins and Purkinje-granule cell interactions in cerebellar development was investigated further in this study by counting the number of Purkinje and granule cells in knockout mutants with a deletion in the proapoptotic gene, Bax. The total number of Purkinje cells was estimated using stereological counting principles in six adult wild type mice, four hemizygous Bax +/- controls, and six Bax -/- knockout mutants. The total number of granule cells per cerebellum was estimated in three adult wild type mice, three hemizygous Bax +/- controls, and three Bax -/- knockout mutants. The number of Purkinje cells increased significantly by over 30% in the Bax -/- knockout mutants compared with wild type and hemizygote controls, whereas the number of granule cells was unchanged in the Bax -/- mutants. There was no change in the volume of the cerebellar cortex or in the size of Purkinje cell bodies in the Bax -/- mutants, implying that Purkinje cell density was increased in the Bax -/- mutants. The increase in Purkinje cell numbers in the Bax -/- knockout mice supports previous evidence that Purkinje cells undergo a period of naturally occurring cell death that is mediated at least in part by the cell death proteins Bcl-2 and Bax. The lack of an effect of Bax gene expression on granule cell numbers indicates that Bax is not an obligate participant in naturally occurring cell death in granule cells. PMID- 11413549 TI - Ventricular proliferation zones in the brain of an adult teleost fish and their relation to neuromeres and migration (secondary matrix) zones. AB - Zones containing actively dividing cells (proliferation zones: PZs), in the brain of adult three-spined sticklebacks, were identified by autoradiographic detection of (3)H-thymidine and immunocytochemical detection of the thymidine analogue 5' bromodeoxyuridine (BrdU), singly or in combination, and by immunocytochemical detection of proliferating cell nuclear antigen (PCNA) by monoclonal antibodies. The PZs are associated with boundaries between adult brain regions, as well as with defined morphofunctional subdivisions. PZs are located at the border between the telencephalon and diencephalon, and at the border between the mesencephalon and the rhombencephalon. In the midbrain, the PZ follows the dorsomedial, caudal, and ventrolateral aspects of each tectal hemisphere, extending over the caudal aspect of the torus semicircularis to the nucleus lateralis valvulae. In the hindbrain, the major PZ apparently represents the persisting embryonic secondary matrix layer of the developing cerebellum. In the forebrain, the PZs are associated with the ventricular zones of the olfactory bulbs and ventral telencephalic area ("subpallium"), dorsal telencephalic area ("pallium"), preoptic region, ventral thalamus, dorsal thalamus, epithalamus, pretectum, posterior tuberculum, and the hypothalamus. The diencephalic PZs are parcellated according to a neuromeric organisation (a synencephalic, a posterior, and an anterior parencephalic neuromere: p1, p2, and p3). The PZs of the secondary prosencephalon (telencephalon and hypothalamus) thus would belong to neuromeres p4-6, but do not form an immediately recognised serial pattern. The prosencephalic PZs correlate well with parts of embryonic migration areas as defined by Bergquist and Kallen ([1954] J. Comp. Neurol. 100:627-659), morphogenetic fields from which postmitotic neurones migrate to their final destination. PMID- 11413550 TI - Ventral mesopontine projections of the caudomedial shell of the nucleus accumbens and extended amygdala in the rat: double dissociation by organization and development. AB - The shell of the nucleus accumbens and central division of the extended amygdala are telencephalic structures that influence motor activity and lately have been regarded by some as components of a single functional-anatomic continuum. Each has a highly differentiated internal organization and output system and distinct pharmacologic responses however, and it is thus likely that each subserves distinct contributions to behavior. In this investigation, nucleus accumbens and extended amygdala outputs were compared by using retrograde tracing in adult and postnatal rats. Fluoro-Gold, when injected into the ventral tegmental area, produced substantial retrograde labeling in the adult nucleus accumbens shell, but only trivial amounts in the central division of the extended amygdala. Injection sites in the lateral mesopontine tegmentum produced robust labeling in the central extended amygdala but little in the nucleus accumbens. The projections of extended amygdala were substantially developed by postnatal day 1, whereas those of the caudomedial shell of the nucleus accumbens only reached the ventral tegmental area by approximately postnatal day 6. Few neurons projecting from the caudomedial shell of the accumbens to the ventral tegmental area were observed even at postnatal day 21. In consideration of the reported importance of the nucleus accumbens, particularly the caudomedial shell, in neural processing related to reward and motivation and the central nervous system response to antipsychotic drugs, it may be important to determine whether processes occurring during the protracted postnatal development of the caudomedial shell are vulnerable to destructive circumstances, such as drug intoxication, maternal separation, or social isolation. PMID- 11413551 TI - Acute effects of combined photodynamic therapy and hyperbaric oxygenation in lung cancer--a clinical pilot study. AB - BACKGROUND AND OBJECTIVE: Photodynamic tumor therapy (PDT) is based upon a photochemical reaction that is limited by the availability of molecular oxygen in the target tissue. The use of hyperbaric oxygenation (HBO) increases the amount of oxygen available for the process may thereby enhance the efficacy of PDT. We investigated the acute effects on tumor stenosis after combined PDT/HBO. PATIENTS AND METHODS: Thirty patients (22 males, 8 females, mean age: 68.8 years; range: 44-78 years) with inoperable non-small cell bronchogenic carcinoma and endobronchial stenosis were studied prospectively. Photosensitization was carried out using a hematoporphyrin-derivative 2 mg/kg BW 48 hours prior to PDT. The light dose was calculated as 300 J/cm fiber tip. The assessment of outcome 1 and 4 weeks after PDT/HBO was performed by endoscopy, chest X-ray, spirometry, laboratory parameters, subjective report of dyspnea, and Karnofsky performance status. RESULTS: At one and four weeks after the treatment, the patients felt a significant improvement of dyspnea and hemoptysis along with an objective subsiding of poststenotic pneumonia, though spirometric parameters revealed no significant difference. A significant reduction of tumor stenosis (P < 0.05) and an improvement of the Karnofsky performance status (P < 0.05) were documented 1 and 4 weeks after PDT/HBO. No therapy related complications were observed. CONCLUSIONS: Although the small number of patients does not allow to draw definitive conclusions to be drawn, the results suggests that combined PDT/HBO represents a new, safe, and technically feasible approach. It enables efficient and rapid reduction of the endoluminal tumor load and helps conditioning the patient for further treatment procedures. PMID- 11413552 TI - Laser treatment of hyperplastic inferior nasal turbinates: a review. AB - BACKGROUND AND OBJECTIVE: Since the early 80s various types of lasers have been used for the reduction of hyperplastic inferior nasal turbinates. Up to now many studies have revealed a variety of important information. To summarize these findings and to determine the value of laser treatment of hyperplastic inferior nasal turbinates, a comparative review of the literature was performed. STUDY DESIGN/MATERIALS AND METHODS: The study of the literature revealed that hyperplastic inferior turbinates of more than 2,000 patients have been treated and followed up. Treatment was performed with the CO2 (10,600 nm), diode (805/810/940 nm), Argon-ion (488/514 nm), KTP (532 nm), Nd:YAG (1,064 nm), and Ho:YAG (2,080 nm) laser in more than 20 studies so far. Generally, the authors of the trials used different laser parameters (power, energy) and application modalities (contact, non-contact, interstitial, superficial). To determine the long-term results objective (active anterior rhinomanometry, acoustic rhinometry, mucociliary function tests, allergy tests) as well as subjective parameters (questionnaire) were recorded and evaluated. In some cases morphological changes of the turbinate tissue were studied by light and scanning electron microscopy (SEM). RESULTS: Laser surgery of inferior turbinates can be performed as an outpatient procedure under local anesthesia. Due to a minimally invasive and controllable coagulation and ablation of soft tissue, almost no complications or bleedings were observed during the operation or postoperatively. Depending on the chosen parameters (power, energy) and the application modalities (contact, non contact, superficial, interstitial) laser treatment of hyperplastic inferior nasal turbinates achieved comparable or better results than most of the conventional techniques for turbinate surgery like conchotomy, electrocautery, cryotherapy, chemical cauterization, and vidian neurectomy. More invasive (radical) operative methods, such as inferior turbinoplasty, submucous turbinectomy, lateral outfracture, partial and total turbinectomy, seemed to be more effective than laser surgery in the long-term. CONCLUSIONS: Laser treatment of hyperplastic inferior nasal turbinates can be considered as a useful, cost effective, and time-saving procedure for the reduction of hyperplastic inferior nasal turbinates. Short operation time, good results, and minor side effects compared to other surgical methods provide an excellent clinical response of the patients. PMID- 11413553 TI - Fluorescence spectroscopic analysis of circulating platelet activation during coronary angioplasty. AB - BACKGROUND AND OBJECTIVE: Platelet activation during percutaneous transluminal coronary angioplasty (PTCA) initiates thrombus formation and plaque regrowth at sites of arterial injury, limiting procedure efficacy. We have developed a simple assay for circulating platelet activation based on fluorescence analysis of membrane fluidity and intracellular calcium concentration and light scattering analysis of platelet aggregation. STUDY DESIGN/MATERIALS AND METHODS: Platelet activation state was measured in 45 patients undergoing angioplasty, before and after treatment with platelet inhibitors. RESULTS: PTCA alone produced a decrease in pyrene dimer formation (P0.0083) and an increase in light scattering at 650 nm (P0.0128). Treatment with ADP and GPIIb/IIIa receptor antagonists reduced PTCA induced changes in pyrene dimer formation. An unexpected decrease in pyrene dimer formation (P0.05) was detected when the GPIIb/IIIa receptor antagonist was given together with an ADP receptor antagonist. CONCLUSIONS: 1) Analysis of membrane fluidity provides a sensitive marker for platelet activation state. 2) Reduced membrane fluidity after combined platelet inhibitor treatments suggests reduced antiplatelet efficacy. PMID- 11413554 TI - Laser solder welding of articular cartilage: tensile strength and chondrocyte viability. AB - BACKGROUND AND OBJECTIVE: The surgical treatment of full-thickness cartilage defects in the knee joint remains a therapeutic challenge. Recently, new techniques for articular cartilage transplantation, such as mosaicplasty, have become available for cartilage repair. The long-term success of these techniques, however, depends not only on the chondrocyte viability but also on a lateral integration of the implant. The goal of this study was to evaluate the feasibility of cartilage welding by using albumin solder that was dye-enhanced to allow coagulation with 808-nm laser diode irradiation. STUDY DESIGN/MATERIALS AND METHODS: Conventional histology of light microscopy was compared with a viability staining to precisely determine the extent of thermal damage after laser welding. Indocyanine green (ICG) enhanced albumin solder (25% albumin, 0.5% HA, 0.1% ICG) was used for articular cartilage welding. For coagulation, the solder was irradiated through the cartilage implant by 808-nm laser light and the tensile strength of the weld was measured. RESULTS: Viability staining revealed a thermal damage of typically 500 m in depth at an irradiance of approximately 10 W/cm(2) for 8 seconds, whereas conventional histologies showed only half of the extent found by the viability test. Heat-bath investigations revealed a threshold temperature of minimum 54 degrees C for thermal damage of chondrocytes. Efficient cartilage bonding was obtained by using bovine albumin solder as adhesive. Maximum tensile strength of more than 10 N/cm(2) was achieved. CONCLUSIONS: Viability tests revealed that the thermal damage is much greater (up to twice) than expected after light microscopic characterization. This study shows the feasibility to strongly laser weld cartilage on cartilage by use of a dye enhanced albumin solder. Possibilities to reduce the range of damage are suggested. PMID- 11413555 TI - A laser-abrasive method for the cutting of enamel and dentin. AB - BACKGROUND AND OBJECTIVE: This paper introduced a new method for the removal of hard dental tissue based upon the use of particles accelerated by laser irradiation, which the authors have called the laser-abrasive method. STUDY DESIGN/MATERIALS AND METHODS: The particles used were sapphire as powder or an aqueous suspension. The effect of the products of enamel ablation was also investigated. The particles were accelerated using submillisecond pulses of Er:YAG and Nd:YAG lasers. A strobing CCD camera was used to measure the speed of the ejected particles. The additional contribution of these particles to the efficiency of laser ablation of enamel and dentin was also investigated. RESULTS: The results showed that the enamel particles produced by the beam-tissue interaction were also accelerated by this process of ablation and were effective in the removal of enamel and dentin. The use of an aqueous suspension of sapphire particles increased the efficiency of enamel removal threefold when compared with the use of an Er:YAG laser with water spray. CONCLUSIONS: The laser-abrasive method allowed for the removal of enamel and dentin at speeds approaching those of the high-speed turbine. PMID- 11413556 TI - Effect of diode laser irradiation on the survival rate of gingival fibroblast cell cultures. AB - BACKGROUND AND OBJECTIVE: The present study is part of a basic research program investigating the cellular effects of the GaAlAs-diode laser with a wavelength of 810 nm on human periodontal tissues. The aim of the investigation was to evaluate the effects of the laser treatment on the survival rate of human gingival fibroblasts (HGF) in monolayer cell culture at different power settings and durations. STUDY DESIGN/MATERIALS AND METHODS: HGF were obtained from a human gingival connective tissue explant. Cells were cultured on 24-well tissue culture plates. One hundred and thirty multi well cell cultures were laser treated. The variable parameters were power output (0.5-2.5 W) and exposure time (60-240 seconds per well). The cultures were analyzed by means of trypan blue staining, and the cell numbers counted under a light microscope. Photographs before and after irradiation were taken and evaluated. RESULTS: Depending on different settings, the laser irradiation caused significant (P < 0.05, t-test) reductions of cell numbers. Exposure time was more relevant to this phenomenon than the power output. Linear regression analysis revealed no unambiguous correlation between power output and cell death, when exposure time was kept constant. CONCLUSIONS: When used for periodontal pocket decontamination, the laser beam may cause damage to collateral periodontal tissues, if the power setting and the duration of the treatment are not adequate. Further investigation, especially with regard to the effects on hard tissue and microorganisms, are needed to give a secure recommendation for irradiation parameters at pocket decontamination. PMID- 11413557 TI - Treatment of nevus of Ota with the Q-switched alexandrite laser. AB - BACKGROUND AND OBJECTIVE: Q-switched laser systems have been shown to be useful for removal of nevus of Ota, a pigmented lesion on the face. The purpose of this study was to evaluate the efficacy of Q-switched alexandrite laser in the treatment of nevus of Ota in 13 patients. STUDY DESIGN/MATERIALS AND METHODS: A spot test was made at 6.0, 7.0, and 8.0 J/cm(2) energy fluence, and the best energy fluence was selected after a 2-month postoperative evaluation. Q-switched alexandrite laser with a mean fluence of 7.290.46 J/cm(2) was used at 8-week intervals. Total treatment ranged from 1 to 15 sessions (mean 7.1 +/- 5.1 sessions). The single shot technique was used. RESULTS: More than 75% lightening was achieved in seven patients, between 51% and 75% in three, less than 50% in one, and less than 25% in one. In two patients with excellent clearance, a very light gray macula in the lower eyelid persisted after 8 and 14 treatment sessions, respectively. Mild transient hypopigmentation that subsided spontaneously after 2 months was observed in one patient. No repigmentation was seen. CONCLUSIONS: The Q-switched alexandrite laser seems to be an effective and safe modality for the treatment of nevus of Ota. PMID- 11413558 TI - Circumscripted pigmentations after iron injections-treatment with Q-switched laser systems. AB - BACKGROUND AND OBJECTIVE: Cutaneous brown discolorations in the area of injection can be a cosmetically bothersome side effect from a parenteral iron treatment, and have not responded to an effective therapy until now. STUDY DESIGN/PATIENTS AND METHODS: We report retrospectively on five patients who were treated with the Q-switched ruby and/or Nd:YAG laser for circumscripted siderosis after intramuscular or intravenous iron injection. RESULTS: The number of required laser treatments was between 3 and 16. The clinical improvement after the first 2 4 sessions was the most impressive, especially after the application of the Nd:YAG laser. Hypopigmentation and scars did not develop. In one case hyperpigmentation appeared. CONCLUSIONS: This study shows that there is an effective method of clearing persistent brown discolorations with few side effects after parenteral iron therapy, using Q-switched laser technology. PMID- 11413559 TI - Laser pulse impact on rat mesenteric blood vessels in relation to laser treatment of port wine stain. AB - BACKGROUND AND OBJECTIVE: To study the impact of laser pulses on animal microvasculature as a model for laser treatment of port wine stains. STUDY DESIGN/MATERIALS AND METHODS: Rat mesenteric blood vessels were irradiated with a laser pulse (585 nm, 0.2-0.6 ms pulse duration, 0.5-30 J/cm(2) radiant exposure). Video microscopy was used to assess vessel dilation, formation of intravascular thrombi, bubble formation, and vessel rupture. Changes in reflection during a laser pulse were measured by simultaneously recording the temporal behavior of the incident and reflected signals. RESULTS: A threshold radiant exposure of approximately 3 J/cm(2) was found for changes in optical properties of blood in vivo, confirming previous in vitro results. Often, laser exposure induced a significant increase in vessel diameter, up to three times the initial diameter for venules and four times for arterioles, within 200 ms after laser exposure. Arterioles were more likely to dilate than venules. Sometimes, immediately after the pulse, round structures, interpreted as being gas bubbles, were seen within the vessel lumen. CONCLUSIONS: A variety of phenomena can occur when blood vessels of sizes comparable to those in port wine stains are irradiated with laser pulses as used in port wine stain treatment. Thrombus formation and vessel rupture have been described before from histological sections of laser-irradiated port wine stains. However, vessel dilation and formation of non-transient gas bubbles as found in this study have not been described before. PMID- 11413560 TI - Cryogen spray cooling in laser dermatology: effects of ambient humidity and frost formation. AB - BACKGROUND AND OBJECTIVE: Dynamics of cryogen spray deposition, water condensation and frost formation is studied in relationship to cooling rate and efficiency of cryogen spray cooling (CSC) in combination with laser dermatologic surgery. STUDY DESIGN/MATERIALS AND METHODS: A high-speed video camera was used to image the surface of human skin during and after CSC using a commercial device. The influence of ambient humidity on heat extraction dynamics was measured in an atmosphere-controlled chamber using an epoxy block with embedded thermocouples. RESULTS: A layer of liquid cryogen may remain on the skin after the spurt termination and prolong the cooling time well beyond that selected by the user. A layer of frost starts forming only after the liquid cryogen retracts. Condensation of ambient water vapor and subsequent frost formation deposit latent heat to the target site and may significantly impair the CSC cooling rate. CONCLUSIONS: Frost formation following CSC does not usually affect laser dosage delivered for therapy of subsurface targets. Moreover, frost formation may reduce the risk of cryo-injury associated with prolonged cooling. The epidermal protection during CSC assisted laser dermatologic surgery can be further improved by eliminating the adverse influence of ambient humidity. PMID- 11413561 TI - An analysis of heat removal during cryogen spray cooling and effects of simultaneous airflow application. AB - BACKGROUND AND OBJECTIVE: Cryogen spray cooling (CSC) is a method used to protect the epidermis from non-specific thermal injury that may occur as a result of various dermatological laser procedures. However, better understanding of cryogen deposition and skin thermal response to CSC is needed to optimize the technique. STUDY DESIGN/MATERIALS AND METHODS: Temperature measurements and video imaging were carried out on an epoxy phantom as well as human skin during CSC with and without simultaneous application of airflow which was intended to accelerate cryogen evaporation from the substrate surface. An inverse thermal conduction model was used to estimate heat flux and total heat removed. RESULTS: Lifetime of the cryogen film deposited on the surface of skin and epoxy phantom lasted several hundred milliseconds beyond the spurt, but could be reduced to the spurt duration by application of airflow. Values over 100 J/cm(3) were estimated for volumetric heat removed from the epidermis using CSC. CONCLUSIONS: "Film cooling" instead of "evaporative cooling" appears to be the dominant mode of CSC on skin. Estimated values of heat removed from the epidermis suggest that a cryogen spurt as long as 200 milliseconds is required to counteract heat generated by high laser fluences (e.g., in treatment of port wine stains) in patients with high concentration of epidermal melanin. Additional cooling beyond spurt termination can be avoided by simultaneous application of airflow, although it is unclear at the moment if avoiding the additional cooling would be beneficial in the actual clinical situation. PMID- 11413562 TI - Group behavioral therapy of obsessive-compulsive disorder: seven vs. twelve-week outcomes. AB - Previous research has demonstrated that individualized behavioral exposure and response prevention therapy is an effective treatment for Obsessive-Compulsive Disorder. In our prior preliminary report, 7-week group exposure and response prevention therapy was also found effective in reducing obsessions and compulsions. The present report describes a larger sample (N=113) of treatment seeking obsessive-compulsives who received group behavioral therapy. As before, group exposure and response prevention significantly improved ratings of obsessions, compulsions, and depression. These improvements were maintained at 3 month and long-term follow-up. A sub-sample of patients who received 12 weeks of treatment had outcomes at the end of the group and at follow-up that did not significantly differ from those who received 7 weeks of treatment. These results confirm the efficacy of a 7-week behavioral treatment program administered in a group format. PMID- 11413563 TI - Reliability and validity of a structured interview guide for the Hamilton Anxiety Rating Scale (SIGH-A). AB - The Hamilton Anxiety Rating Scale, a widely used clinical interview assessment tool, lacks instructions for administration and clear anchor points for the assignment of severity ratings. We developed a Structured Interview Guide for the Hamilton Anxiety Scale (SIGH-A) and report on a study comparing this version to the traditional form of this scale. Experienced interviewers from three Anxiety Disorders research sites conducted videotaped interviews using both traditional and structured instruments in 89 participants. A subset of the tapes was co-rated by all raters. Participants completed self-report symptom questionnaires. We observed high inter-rater and test-retest reliability using both formats. The structured format produced similar but consistently higher (+ 4.2) scores. Correlation with a self-report measure of overall anxiety was also high and virtually identical for the two versions. We conclude that in settings where extensive training is not practical, the structured scale is an acceptable alternative to the traditional Hamilton Anxiety instrument. PMID- 11413564 TI - Factorial validity of the Childhood Trauma Questionnaire in men and women. AB - In an effort to confirm the factorial validity of the Childhood Trauma Questionnaire (CTQ) across sex, the items from the CTQ for 916 university students were subjected to confirmatory factor analysis. Results indicated that the factor structure for the CTQ was significantly different for men and women. For women, the items from the Physical Abuse subscale did not create a stable factor and thus appear not to be conceptually valid. Conversely, for men, the five-factor model provided a relatively good fit to the data. This investigation provides important information regarding sex differences in the factorial validity of the CTQ. Implications and future research directions are discussed. PMID- 11413565 TI - Anxiety across the life span: epidemiological evidence and treatment data. PMID- 11413566 TI - Prevalence of childhood ADHD among patients with Generalized Anxiety Disorder and a comparison condition, Social Phobia. PMID- 11413567 TI - Quality of life following cognitive behavioral treatment for social anxiety disorder: preliminary findings. PMID- 11413568 TI - Community correlates of outcomes in subjects with panic attacks. AB - The purpose of this study was to document the association between community factors and mental health outcomes in subjects with panic attacks. Randomly selected adults from 18 census tracts were screened for the presence of panic attacks. A structured interview was used to assess health care utilization, psychiatric morbidity, quality of life, and sense of control over panic. Community measures were obtained from census data. Regression analyses found that each community measure was associated with at least one outcome even when adjusted for individual socioeconomic status and barriers to access. Research concerning mental health outcomes in subjects with panic attacks should include community-level data. PMID- 11413570 TI - How did we ever get here? PMID- 11413571 TI - Hey, "sleeping giant," wake up and smell the money you're losing! PMID- 11413572 TI - The gift you were given. PMID- 11413573 TI - Maxillofacial prosthetics. PMID- 11413574 TI - Treatment of fremitus. PMID- 11413575 TI - The University of Minnesota Oral Health Clinical Research Center. PMID- 11413576 TI - Setting a fair fee: a strategic process. PMID- 11413577 TI - Recognizing the best in everyone. PMID- 11413578 TI - The Minnesota Board of Dentistry: real people, real issues. PMID- 11413579 TI - The sun and skin cancer. What you need to know about diagnosis and treatment. AB - Skin cancer is the most common malignancy in man, with an incidence of 1,000,000 new cases diagnosed in the United States in 1999. There are three recognized types of skin cancer: basal cell carcinoma, squamous carcinoma, and melanoma. These cancers frequently occur on the sun-exposed areas (face, ears, scalp, etc.) of the head and neck. These areas are amenable to examination by general dentists during routine oral examinations. Early diagnosis of skin cancers and subsequent treatment result in high cure rates. Dentists are in a unique position to help in the diagnosis of skin cancer in their patients due to their frequent contact with patients compared to other health care practitioners. PMID- 11413580 TI - Amalgam waste management: issues and answers. AB - In reviewing U.S. dental amalgam waste management activities, one finds a wide variety of approaches, processes, and results, within which the authors have observed an inconsistency of definitions, measurements, standards, and even of clearly applied goals. As a result, outcomes have varied greatly from community to community. This manuscript attempts to deliver logical options which might prevail even within an atmosphere of differing perspectives. The authors share the belief that successful outcomes are achievable within a collaborative environment of shared learning and cooperative planning. By example, the authors attempt to demonstrate how the joint acceptance of certain definitions and goals has served as a pathway to success in one community's dental waste management project. The goal of this paper is to share tools that may contain some measure of applicability to other similar collaborations. PMID- 11413581 TI - Matters of choice. The Presidents' Forum 2000. PMID- 11413582 TI - Antibiotic abuse; let's stop it now. PMID- 11413583 TI - Clinical pathology conference. Mucous membrane pemphigoid. PMID- 11413584 TI - Functioning adrenal tumours in children and adolescents: an institutional experience. AB - BACKGROUND: The purpose of the present paper was to carry out an audit of clinicopathological profile and treatment outcome in 13 children with functioning adrenal tumours. METHODS: The medical records of 13 children with functioning adrenal tumours who were managed between June 1990 and January 1999 were reviewed. Demographic data, clinical features, biochemical and localization studies, operative details and follow-up records were studied. Children with neuroblastoma were excluded. RESULTS: The mean age was 7.4 +/- 5.3 years. Seven patients had Cushing's syndrome (CS), two patients had virilizing tumours, three patients had phaeochromocytoma (PCC) and one patient had Conn's syndrome. All patients (except one child with CS) were treated surgically. Two children with adrenocortical carcinoma (ACCa) died during the perioperative period. Histopathological diagnosis was adrenal cortical adenoma (ACAd) in four patients, ACCa in five patients and PCC in three patients. Two ACCa patients died of metastases at 12 and 14 months, respectively, while the third is alive and well at 30 months. Children with ACAd are alive and well at 91, 56, 32 and 27 months postoperatively. Children with PCC are free of disease (normal urinary metanephrines) at 63, 18 and 8 months after surgery but require antihypertensive drugs in low doses. CONCLUSION: The outcome of surgery is good in cases of ACAd and PCC. Although outcome is poor in ACCa, surgery remains the mainstay of treatment and offers good palliation. PMID- 11413585 TI - The development of undergraduate curricula in surgery: II. Generic surgery. AB - The present paper focuses upon the issues in curricular reform that have specific relevance for surgeons. A central theme is that, taking into account the dual diminution of general surgery and large central teaching hospitals, there is a need to have a clear vision of what should be included in surgical curricula and how we can adjust to new methods of teaching and learning. PMID- 11413586 TI - 'Philanthropy': address to council members of The Royal Australasian College of Surgeons. PMID- 11413587 TI - Thyroid surgery and voice-related outcomes. PMID- 11413588 TI - Ulcers in diabetic feet: an orthopaedic not a vascular problem. PMID- 11413589 TI - Ulcers in diabetic feet: an orthopaedic not a vascular problem. PMID- 11413590 TI - Audit of outpatient laparoscopic cholecystectomy. PMID- 11413591 TI - Neonatal acute haemorrhagic gastritis and antenatal exposure to indomethacin for tocolysis. PMID- 11413592 TI - Low anterior resection and colonic J-pouch in the treatment of adult Hirschsprung's disease. PMID- 11413593 TI - Impalement injury of the thorax. PMID- 11413594 TI - Persistent synovial fistula after arthroscopy in carbon synovitis. PMID- 11413595 TI - Recurrent basal cell carcinoma causing spinal cord compression. PMID- 11413596 TI - Jack Smyth: a major contribution to surgery. AB - This personal comment records the contributions of the late Dr John Smyth and Royal Newcastle Hospital to surgical audit and surgical training. Smyth introduced surgical audit to this country against considerable professional opposition. The first publication on the subject appeared in The Medical Journal of Australia in 1959. PMID- 11413597 TI - Successful ex vivo renal artery reconstruction and renal autotransplantation. AB - BACKGROUND: The increasing experience with renal allotransplantation has led to continuing development in vascular surgical techniques. These improvements have enabled complex ex vivo renal artery surgery and renal autotransplantation to be performed. The aims of the present study were to describe the results achieved with renal autotransplantation and ex vivo renal artery reconstruction (RAR) at the Newcastle Transplant Unit, John Hunter Hospital, and to review the current indications for such surgery. METHODS: A retrospective review was performed of patients who required renal autotransplantation with or without RAR at John Hunter Hospital, between 1991 and 1999. Data were obtained from the Newcastle Transplant Unit and the Medical Record Department of John Hunter Hospital. RESULTS: Two patients required ex vivo RAR and renal autotransplantation for severe fibromuscular dysplasia (FMD) complicated by stenoses and renal artery branch aneurysms. The third patient required autotransplantation for bilateral retroperitoneal fibrosis. There was one postoperative complication of pelviureteric junction obstruction that was treated successfully with a temporary ureteric stent. All patients demonstrated normal graft function and were normotensive on follow up, which ranged from 2.5 to 5 years. CONCLUSION: The present review confirms the long-term benefits of ex vivo RAR and renal autotransplantation that have been demonstrated by previous studies. In transplant units experienced with this surgery it has been shown to be a successful and durable technique for the treatment of a variety of vascular, urologic and other diseases. PMID- 11413598 TI - Trauma training in Australia and New Zealand: results of a survey of advanced surgical trainees. AB - BACKGROUND: The surgical management of trauma within Australia and New Zealand has recently been undergoing major organizational changes. The aim of the present paper was to evaluate the attitudes and experience of Australian and New Zealand advanced surgical trainees in this changing climate and to identify problems with trauma training. METHODS: A survey assessing important areas of trauma management and training was sent to all advanced surgical trainees of the Royal Australasian College of Surgeons. RESULTS: Two hundred and seventy-two of 587 trainees responded (46%). Overall 85% of trainees believed they would be involved in trauma management in the future. The majority of trainees reported low rates of involvement and consultant supervision in trauma resuscitations. Only 32% of general surgical trainees believed that their exposure to major trauma operations was very adequate despite an average of 12.3 trauma operations per year. Seventy per cent of general surgical trainees reported a very adequate level of consultant supervision at trauma operations. In contrast 86% of orthopaedic trainees reported a very adequate exposure to trauma operations with an average of 221 orthopaedic trauma operations per year. Only 46% of orthopaedic trainees reported a very adequate level of consultant supervision at trauma operations. CONCLUSIONS: Regional rotations may need to be developed to even out trainees' experience in trauma management. The low level of supervision in trauma resuscitations and orthopaedic surgical training requires attention. This survey warrants repeating in a prospective manner. PMID- 11413599 TI - Surgical treatment of cerebral ischaemia caused by cervical arterial lesions due to Takayasu's arteritis: preliminary results of 49 cases. AB - BACKGROUND: The present paper reports the preliminary experience with surgical treatment of 49 cases of cerebral ischaemia caused by cervical arterial lesions due to Takayasu's arteritis (TA). METHODS: Six men and 43 women were treated between June 1992 and October 1997. Their ages ranged from 15 to 42 years, with a mean of 26.5 years. Duration of symptoms was from 10 days to 8 years with a mean of 32 months. Bypasses from the ascending aorta to the axillary or subclavian artery and to the carotid artery were performed in 31 cases. Subclavian-to carotid bypass was performed in six cases. Percutaneous transluminal angioplasty (PTA) was used in four cases. RESULTS: An excellent result was achieved in 28.6% of cases, a fair result was achieved in 36.7% of cases, improvement was achieved in 22.5% of cases, 2.0% of cases were unchanged, and in 10.2% of cases death occurred before discharge. During a mean follow up of 38 months these values were 26.2%, 42.8%, 16.7%, 4.8%, and 2.4%, respectively; recurrence occurred in 7.1% of cases. CONCLUSION: When cerebral perfusion is jeopardized, a definitive corrective procedure is advised at a time when the patient is in a relatively stable condition. Percutaneous transluminal angioplasty is the first choice although the recurrence rate is very high, and complications such as arterial rupture and thrombosis formation are serious. Patients with occlusive lesions of all four cervical arteries usually have severe cerebral ischaemia and their distal runoff is usually unable to be visualized on angiography. But it was found that in 95% of cases the internal carotid artery is still patent (confirmed by exploration), therefore an ascending aorta-to-carotid bypass is feasible in most instances. Unfortunately, cerebral re-perfusion syndrome is a serious unresolved problem. PMID- 11413600 TI - Feasibility of day-care open cholecystectomy: evaluation in an inpatient model. AB - BACKGROUND: Open cholecystectomy is still a fairly frequently performed operation worldwide, and is used where laparoscopic surgery has failed or is contraindicated, and where some surgeons do not operate laparoscopically for technical reasons. In developing countries laparoscopic cholecystectomy is costly and is available only in a few centres. The present study was conducted to assess the feasibility of day-care open cholecystectomy using an inpatient model. METHODS: Thirty patients were subjected to open cholecystectomy. Intraoperative nasogastric decompression and local wound infiltration with 0.25% bupivacaine was carried out. Postoperatively patients were encouraged to be ambulant, pass urine and start oral fluids. Intravenous fluids and parenteral medication were stopped at 8 p.m. when patients were assessed for feasibility for discharge. All patients were reassessed the next morning for any adverse effects that could have occurred had the patients been discharged on the evening of surgery. RESULTS: Prospectively, 73.3% of patients were considered to be dischargeable on the evening of surgery; but on reassessment the next morning, retrospectively, 93.3% of patients were actually dischargeable on the evening of surgery. A total of 76.6% of patients was actually discharged within 24 h of surgery. No patient required readmission. On follow up there were no complications that could be attributable to early discharge. A total of 83.3% of patients approved of day care open cholecystectomy. CONCLUSIONS: Day-care open cholecystectomy is safe and feasible. In developing countries, where the use of laparoscopic surgery is limited due to resource constraints, day-care open cholecystectomy can lead to substantial savings in health-care resources. PMID- 11413601 TI - Erythromycin enhances oesophageal motility in patients with gastro-oesophageal reflux. AB - BACKGROUND: Intravenous (i.v.) erythromycin enhances gastric emptying and oesophageal motility in both healthy and disease situations, acting either as a motilin or acetylcholine agonist. The purpose of the present paper was to investigate any possible effect of i.v. erythromycin on oesophageal motility in patients with gastro-oesophageal reflux (GOR). METHODS: In 15 patients with GOR (proven on 24-h ambulatory oesophageal pH measurement), standard oesophageal manometry was performed after i.v. injection of placebo and 200 mg erythromycin, in a random blind fashion. RESULTS: Erythromycin significantly increased lower oesophageal sphincter (LOS) pressure from 17 +/- 5 to 41 +/- 10 mmHg (P < 0.001), without affecting the postdeglutition relaxation of LOS. Erythromycin also increased the amplitude (from 79 +/- 34 to 97 +/- 40 mmHg; P < 0.001), duration (from 3.4 +/- 0.6 to 3.8 +/- 0.6 s; P = 0.005), velocity (from 3.1 +/- 0.8 to 3.5 +/- 1.15 cm/s; P = 0.0047) and strength (from 149 +/- 84 to 201 +/- 103 mmHg.s; P < 0.001) of peristalsis at 5 cm proximal to the LOS. Similarly, the drug increased the amplitude of peristalsis at 10 and 15 cm proximal to the LOS (from 70 +/- 39 to 77.4 +/- 37 mmHg; P = 0.049 and from 36 +/- 20 to 49 +/- 36 mmHg; P = 0.004, respectively) and the duration of peristalsis at the same levels (from 3.1 +/- 0.6 to 3.3 +/- 0.5 s; P = 0.011, and from 2.7 +/- 0.6 to 3 +/- 0.5 s; P = 0.003, respectively). CONCLUSION: Intravenously administered erythromycin improves impaired oesophageal motility in patients with GOR. This observation might be of clinical use. PMID- 11413602 TI - Old dog learns new trick. PMID- 11413603 TI - Access to dental care--the ongoing discussion. PMID- 11413604 TI - Interdisciplinary care for persons with cleft lip and palate in the year 2001. PMID- 11413605 TI - Pop! Goes your big smile! PMID- 11413607 TI - The journey back. Part one. PMID- 11413606 TI - A crisis in the community. PMID- 11413608 TI - Guided tissue regeneration in periodontal therapy. AB - In teeth where continued function requires additional periodontal support, optimal treatment requires not only controlling periodontal infection, but also regeneration of periodontal support lost to periodontal disease. Today, guided tissue regeneration (GTR) is a technique with significant clinical and histologic documentation of periodontal regeneration. This paper will address GTR barriers and clinical indications for using these materials to successfully regenerate the periodontium. PMID- 11413609 TI - Development of a classification system for periodontal diseases and conditions. AB - Classification systems are necessary in order to provide a framework in which to scientifically study the etiology, pathogenesis, and treatment of diseases in an orderly fashion. In addition, such systems give clinicians a way to organize the health care needs of their patients. The last time scientists and clinicians in the field of periodontology and related areas agreed upon a classification system for periodontal diseases was in 1989 at the World Workshop in Clinical Periodontics. Subsequently, a simpler classification was agreed upon at the 1st European Workshop in Periodontology. These classification systems have been widely used by clinicians and research scientists throughout the world. Unfortunately, the 1989 classification had many shortcomings, including: (1) considerable overlap in disease categories, (2) absence of a gingival disease component, (3) inappropriate emphasis on age of onset of disease and rates of progression, and (4) inadequate or unclear classification criteria. The 1993 European classification lacked the detail necessary for adequate characterization of the broad spectrum of periodontal diseases encountered in clinical practice. The need for a revised classification system for periodontal diseases was emphasized during the 1996 World Workshop in Periodontics. In 1997 the American Academy of Periodontology responded to this need and formed a committee to plan and organize an international workshop to revise the classification system for periodontal diseases. The proceedings in this volume are the result of this reclassification effort. The process involved development by the Organizing Committee of an outline for a new classification and identification of individuals to write state-of-the-science reviews for each of the items on the outline. The reviewers were encouraged to depart from the preliminary outline if there were data to support any modifications. On October 30-November 2, 1999, the International Workshop for a Classification of Periodontal Diseases and Conditions was held and a new classification was agreed upon (Figure 1). This paper summarizes how the new classification for periodontal diseases and conditions presented in this volume differs from the classification system developed at the 1989 World Workshop in Clinical Periodontics. In addition, an analysis of the rationale is provided for each of the modifications and changes. PMID- 11413610 TI - The grand design. PMID- 11413611 TI - Chronic orofacial pain disorders: developments in diagnosis and treatment include proponents of specialization. PMID- 11413612 TI - Treading water: Minnesota's dental workforce in the year 2000. PMID- 11413613 TI - The graying of America: challenges and opportunities for the dental industry. PMID- 11413614 TI - Non-odontogenic toothache. AB - Toothache is a common complaint in the dental office. Most toothaches have their origin in the pulpal tissues of periodontal structures. These odontogenic pains are managed well and predictably by dental therapies. Non-odontogenic toothaches are often difficult to identify and can challenge the diagnostic ability of the clinician. The most important step toward proper management of toothache is to be suspicious that the pain may not be of dental origin. The cardinal warning symptoms of non-odontogenic toothache are as follow: A. Spontaneous multiple toothaches. B. Inadequate local dental cause for the pain. C. Stimulating, burning, non-pulsatile toothaches. D. Constant, unremitting, non-variable toothaches. E. Persistent, recurrent toothaches. F. Local anesthetic blocking of the offending tooth does not eliminate the pain. G. Failure of the toothache to respond to reasonable dental therapy. PMID- 11413615 TI - School of Dentistry's new dean charts cutting edge course. PMID- 11413616 TI - How can something so good be such a pain? PMID- 11413617 TI - Burning mouth syndrome: diagnosis and treatment. PMID- 11413618 TI - Achieving clinical excellence. AB - Clinical excellence can be divided into three defining categories: procedures, systems, and communications. Six steps have been introduced to guide our journey. New systems will produce less stressful, more profitable dentistry, and with this commitment to clinical excellence, patient care will improve automatically. Evaluating our practices and making the necessary changes will bring significant rewards. In Designing the Future, Visions and Strategies, I state, "Maximum efficiency, profitability, and security, the rewards of business momentum, come only to those who start paddling early. To avoid dead water, we must view business strategies in the context of social, technological, and economic currents. It's the difference between actively engaging trends and being destroyed by them." PMID- 11413619 TI - Winkleman's conundrum. PMID- 11413620 TI - The State of an Art. PMID- 11413621 TI - Reason why quitters are winners. PMID- 11413622 TI - New concepts in dental adhesion. PMID- 11413623 TI - RM matters: understanding cultural differences. PMID- 11413624 TI - Incipient speciation in sympatric Nicaraguan crater lake cichlid fishes: sexual selection versus ecological diversification. AB - The growing body of empirical evidence for sympatric speciation has been complemented by recent theoretical treatments that have identified evolutionary conditions conducive to speciation in sympatry. The Neotropical Midas cichlid (Amphilophus citrinellum) fits both of the key characteristics of these models, with strong assortative mating on the basis of a colour polymorphism coupled with trophic and ecological differentiation derived from a polymorphism in their pharyngeal jaws. We used microsatellite markers and a 480 bp fragment of the mitochondrial DNA control region to study four polymorphic populations of the Midas cichlid from three crater lakes and one large lake in Nicaragua in an investigation of incipient sympatric speciation. All populations were strongly genetically differentiated on the basis of geography. We identified strong genetic separation based on colour polymorphism for populations from Lake Nicaragua and one crater lake (Lake Apoyo), but failed to find significant genetic structuring based on trophic differences and ecological niche separation in any of the four populations studied. These data support the idea that sexual selection through assortative mating contributes more strongly or earlier during speciation in sympatry than ecological separation in these cichlids. The long term persistence of divergent cichlid ecotypes (as measured by the percentage sequence divergence between populations) in Central American crater lakes, despite a lack of fixed genetic differentiation, differs strikingly from the patterns of extremely rapid speciation in the cichlids in Africa, including its crater lakes. It is unclear whether extrinsic environmental factors or intrinsic biological differences, e.g. in the degree of phenotypic plasticity, promote different mechanisms and thereby rates of speciation of cichlid fishes from the Old and New Worlds. PMID- 11413625 TI - Ornamental plumage does not signal male quality in red-billed queleas. AB - Sexually selected ornaments often function as condition-dependent signals of quality (or 'indicators'). When ornamentation is costly, only high-quality individuals can afford to produce the most elaborate signals. The plumage ornamentation of male red-billed queleas, Quelea quelea, is an ideal candidate for an indicator because it is continuously variable, conspicuous, sexually dimorphic, is displayed only during breeding and is partially based on carotenoid pigmentation. However, I show here that quelea plumage is not an indicator because first, plumage colour is not correlated with physical condition or age; second, plumage colour is a genetically determined phenotype that is unresponsive to environmental variation; third, different plumage characters have bimodal distributions; fourth, plumage characters vary independently of one another; and finally, plumage colour is not correlated with reproductive success. To my knowledge, this is the first demonstration of non-condition dependence in colourful and sexually dimorphic breeding ornamentation. Instead, plumage variation may function as a sexually selected signal of individual identity among territorial males that nest in huge, densely packed and highly synchronized colonies. PMID- 11413626 TI - Male and female mate choice affects offspring quality in a sex-role-reversed pipefish. AB - Where both sexes invest substantially in offspring, both females and males should discriminate between potential partners when choosing mates. The degree of choosiness should relate to the costs of choice and to the potential benefits to be gained. We measured offspring quality from experimentally staged matings with preferred and non-preferred partners in a sex-role-reversed pipefish, Syngnathus typhle L. Here, a substantial male investment in offspring results in a lower potential reproductive rate in males than in females, and access to males limits female reproductive success rather than vice versa. Thus, males are choosier than females and females compete more intensely over mates than do males. Broods from preferred matings were superior at escaping predation, when either males or females were allowed to choose a partner. However, only 'choosing' females benefited in terms of faster-growing offspring. Our results have important implications for mate-choice research: here we show that even the more competitive and less choosy sex may contribute significantly to sexual selection through mate choice. PMID- 11413627 TI - Size-dependent response to conspecific mating calls by male crickets. AB - Male sexual displays provide females with information that is crucial to their reproductive decisions. That same information is available to eavesdroppers, with potential consequences for both signaller and receiver. We present empirical evidence for size-dependent responses to intersexual communication by conspecific rivals. Acoustic features of a male house cricket's (Acheta domesticus) mating call are positively associated with its size, with females preferring the calls of larger males. In order to investigate whether conspecific males make use of the information content of mating calls, we examined their phonotactic responses to call recordings that differ in attractiveness to females. Males of all sizes exhibited positive phonotaxis, with smaller males showing a clear preference for female-preferred calls. Smaller males were also less likely to seek contact with the speaker playing their chosen call. We discuss possible explanations for this size-dependent male behaviour. PMID- 11413628 TI - Clutch size evolution under sexual conflict enhances the stability of mating systems. AB - Models of optimal clutch size often implicitly assume a situation with uniparental care. However, the evolutionary conflict between males and females over the division of parental care will have a major influence on the evolution of clutch size. Since clutch size is a female trait, a male has little possibility of directly influencing it. However, the optimal clutch size from a female's perspective will depend on the amount of paternal care her mate is expected to provide. The sexual conflict over parental care will in its turn be affected by clutch size, since a larger clutch makes male care more valuable. Hence, there will be joint evolution of mating system and clutch size. In this paper, we demonstrate that this joint evolution will tend to stabilize the mating system. In a situation with conventional sex roles, this joint evolution might result in either increased clutch size and biparental care or reduced clutch size and uniparental female care. Under some circumstances the initial conditions might determine which will be the outcome. These results demonstrate that it may be difficult to deduce whether biparental care evolved because of few opportunities for breeding males increasing their fitness by attracting additional mates or because of the importance of male care for offspring fitness by studying prevailing mating systems using, for example, male removals or manipulation of males' opportunities for finding additional mates. In general terms, we demonstrate that models of life-history evolution have to consider the social context in which they evolve. PMID- 11413629 TI - Brood desertion by female shorebirds: a test of the differential parental capacity hypothesis on Kentish plovers. AB - The aim of this study was to examine whether the energetic costs of reproduction explain offspring desertion by female shorebirds, as is suggested by the differential parental capacity hypothesis. A prediction of the hypothesis is that, in species with biparental incubation in which females desert from brood care after hatching, the body condition of females should decline after laying to a point at which their body reserves are too low for continuing parental care. We tested this prediction on Kentish plovers (Charadrius alexandrinus) in which both sexes incubate but the females desert from brood care before the chicks fledge. We found no changes in either the body masses or body compositions of both individual male and female plovers from early incubation and throughout early chick rearing. Furthermore, the timing of brood desertion by females was not affected by their body condition. Neither did we find gender differences in the energetic costs of incubation. There were no differences in the timing of brood desertion between experimental and control females in an experiment in which we lengthened or shortened the duration of incubation by one week. These results indicate that energetic costs do not explain offspring desertion by female Kentish plovers and that the needs of chicks for parental care rather than cumulative investment by females is what determines the timing of brood desertion. PMID- 11413630 TI - The spatial ultimatum game. AB - In the ultimatum game, two players are asked to split a certain sum of money. The proposer has to make an offer. If the responder accepts the offer, the money will be shared accordingly. If the responder rejects the offer, both players receive nothing. The rational solution is for the proposer to offer the smallest possible share, and for the responder to accept it. Human players, in contrast, usually prefer fair splits. In this paper, we use evolutionary game theory to analyse the ultimatum game. We first show that in a non-spatial setting, natural selection chooses the unfair, rational solution. In a spatial setting, however, much fairer outcomes evolve. PMID- 11413631 TI - Host-parasite coevolution in a multilocus gene-for-gene system. AB - This paper examines a mathematical model for the coevolution of parasite virulence and host resistance under a multilocus gene-for-gene interaction. The degrees of parasite virulence and host resistance show coevolutionary cycles for sufficiently small costs of virulence and resistance. Besides these coevolutionary cycles of a longer period, multilocus genotype frequencies show complex fluctuations over shorter periods. All multilocus genotypes are maintained within host and parasite classes having the same number of resistant/virulent alleles and their frequencies fluctuate with approximately equally displaced phases. If either the cost of virulence or the number of resistance loci is larger then a threshold, the host maintains the static polymorphism of singly (or doubly or more, depending on the cost of resistance) resistant genotypes and the parasite remains universally avirulent. In other words, host polymorphism can prevent the invasion of any virulent strain in the parasite. Thus, although assuming an empirically common type of asymmetrical gene for-gene interaction, both host and parasite populations can maintain polymorphism in each locus and retain complex fluctuations. Implications for the red queen hypothesis of the evolution of sex and the control of multiple drug resistance are discussed. PMID- 11413632 TI - Phylogeography and regional endemism of a passively dispersing zooplankter: mitochondrial DNA variation in rotifer resting egg banks. AB - We investigated the phylogeography of the salt water rotifer Brachionus plicatilis, a cyclical parthenogen with passive dispersal mechanisms, using resting eggs recovered from saline lake sediments. Individual resting eggs were obtained from a large selection of lakes which were representative of five endorheic basins and the chain of coastal ponds in the Iberian Peninsula. The novel use of resting eggs allows the integration of seasonal and annual variations as well as the impact of stochastic effects such as drift and local extinction. A 653 bp fragment of the mitochondrial cytochrome oxidase subunit I (COI) gene was sequenced from 98 eggs. Our results revealed a deep phylogeographical structure in this species, with a division into two main lineages with distinct geographical distributions, which probably diverged at the beginning of the Pleistocene period. Most of the mitochondrial DNA haplotypes were restricted to single lakes. Nested clade analysis supported Early Pleistocene fragmentation of populations, low gene flow and some long-distance colonization. These conclusions contrast strongly with previous ideas on rotifer biogeography and this pattern is consistent with a recolonization of the Iberian Peninsula from two glacial refugia. The results provide new insights into the processes responsible for the genetic diversification of passive dispersers, a life-history trait typical of zooplanktonic biotas. PMID- 11413633 TI - Tracking colonization and diversification of insect lineages on islands: mitochondrial DNA phylogeography of Tarphius canariensis (Coleoptera: Colydiidae) on the Canary Islands. AB - The genus Tarphius Erichson (Coleoptera: Colydiidae) is represented by 29 species on the Canary Islands. The majority are rare, single-island endemics intimately associated with the monteverde (laurel forest and fayal-brezal). The Tarphius canariensis complex is by far the most abundant and geographically wide-spread, occurring on Gran Canaria, Tenerife and La Palma. Eighty-seven individuals from the T. canariensis complex were sequenced for 444 bp of the mitochondrial DNA cytochrome oxidase I gene (COI), 597 bp of the COII gene and the intervening tRNA(leu) gene. A neighbour-joining analysis of maximum-likelihood distances put La Palma as a single monophyletic clade of haplotypes occurring within a larger clade comprising all Tenerife haplotypes. Gran Canarian haplotypes were also monophyletic occurring on a separate lineage. Using a combination of the phylogeographic pattern for T. canariensis, geological data, biogeography of the remaining species and estimated divergence times, we proposed a Tenerifean origin in the old Teno massif and independent colonizations from here to north-eastern Tenerife (Anaga), Gran Canaria and La Palma. New methods of estimating diversification rates using branching times were applied to each island fauna. All islands exhibited a gradually decreasing rate of genetic diversification similar to that seen for Brachyderes rugatus (Coleoptera: Curculionidae) from the Canary Islands. PMID- 11413634 TI - The effects of metapopulation structure on indirect interactions in host parasitoid assemblages. AB - The interaction between two species that do not compete for resources but share a common natural enemy is known as apparent competition. In the absence of other limiting factors, such three-species interactions are impermanent, with one species being excluded from the assemblage by the natural enemy. Here, the effects of metapopulation structure are explored in a system of two hosts that experience apparent competition through a shared parasitoid. A coupled-map lattice model is developed and used to explore species coexistence and patterns of patch occupancy at the metapopulation scale. Linking local and regional dynamics favours coexistence by uncoupling the dynamics of the three species in space. Coexistence is promoted by the inferior species being either a fugitive or a sedentary species. The occurrence of these two mutually exclusive mechanisms of coexistence is influenced by the relative dispersal of the inferior apparent competitor. PMID- 11413635 TI - Activity synchrony and social cohesion: a fission-fusion model. AB - A social group can only be spatially coherent if its members synchronize activities such as foraging and resting. However, activity synchronization is costly to individuals if it requires them to postpone an activity that would be personally more profitable in order to do what the rest of the group is doing. Such costs will be particularly high in groups whose members belong to different age, size or sex classes since the optimal allocation of time to various activities is likely to differ between such classes. Thus, differences in the costs of activity synchronization between and within classes could cause non homogenous groups to be less stable than homogenous groups, with the result that homogenous groups predominate in the population: that is, they could cause 'social segregation' of animals of different sex, size or age. We develop a model that predicts the degree of social segregation attributable to differences in activity synchronization between homogenous and non-homogenous groups and use this model in determining whether activity synchronization can explain intersexual social segregation in red deer (Cervus elaphus). Differences in activity synchronization between mixed-sex and unisex groups of red deer explained 35% of the observed degree of intersexual social segregation, showing that activity synchronization is an important cause of social segregation in this species. PMID- 11413636 TI - Bubonic plague: a metapopulation model of a zoonosis. AB - Bubonic plague (Yersinia pestis) is generally thought of as a historical disease; however, it is still responsible for around 1000-3000 deaths each year worldwide. This paper expands the analysis of a model for bubonic plague that encompasses the disease dynamics in rat, flea and human populations. Some key variables of the deterministic model, including the force of infection to humans, are shown to be robust to changes in the basic parameters, although variation in the flea searching efficiency, and the movement rates of rats and fleas will be considered throughout the paper. The stochastic behaviour of the corresponding metapopulation model is discussed, with attention focused on the dynamics of rats and the force of infection at the local spatial scale. Short-lived local epidemics in rats govern the invasion of the disease and produce an irregular pattern of human cases similar to those observed. However, the endemic behaviour in a few rat subpopulations allows the disease to persist for many years. This spatial stochastic model is also used to identify the criteria for the spread to human populations in terms of the rat density. Finally, the full stochastic model is reduced to the form of a probabilistic cellular automaton, which allows the analysis of a large number of replicated epidemics in large populations. This simplified model enables us to analyse the spatial properties of rat epidemics and the effects of movement rates, and also to test whether the emergent metapopulation behaviour is a property of the local dynamics rather than the precise details of the model. PMID- 11413638 TI - The Noh mask effect: vertical viewpoint dependence of facial expression perception. AB - Full-face masks, worn by skilled actors in the Noh tradition, can induce a variety of perceived expressions with changes in head orientation. Out-of-plane rotation of the head changes the two-dimensional image characteristics of the face which viewers may misinterpret as non-rigid changes due to muscle action. Three experiments with Japanese and British viewers explored this effect. Experiment 1 confirmed a systematic relationship between vertical angle of view of a Noh mask and judged affect. A forward tilted mask was more often judged happy, and one backward tilted more often judged sad. This effect was moderated by culture. Japanese viewers ascribed happiness to the mask at greater degrees of backward tilt with a reversal towards sadness at extreme forward angles. Cropping the facial image of chin and upper head contour reduced the forward-tilt reversal. Finally, the relationship between head tilt and affect was replicated with a laser-scanned human face image, but with no cultural effect. Vertical orientation of the head changes the apparent disposition of facial features and viewers respond systematically to these changes. Culture moderates this effect, and we discuss how perceptual strategies for ascribing expression to familiar and unfamiliar images may account for the differences. PMID- 11413637 TI - Blind subjects construct conscious mental images of visual scenes encoded in musical form. AB - Blind (previously sighted) subjects are able to analyse, describe and graphically represent a number of high-contrast visual images translated into musical form de novo. We presented musical transforms of a random assortment of photographic images of objects and urban scenes to such subjects, a few of which depicted architectural and other landmarks that may be useful in navigating a route to a particular destination. Our blind subjects were able to use the sound representation to construct a conscious mental image that was revealed by their ability to depict a visual target by drawing it. We noted the similarity between the way the visual system integrates information from successive fixations to form a representation that is stable across eye movements and the way a succession of image frames (encoded in sound) which depict different portions of the image are integrated to form a seamless mental image. Finally, we discuss the profound resemblance between the way a professional musician carries out a structural analysis of a musical composition in order to relate its structure to the perception of musical form and the strategies used by our blind subjects in isolating structural features that collectively reveal the identity of visual form. PMID- 11413639 TI - The importance of leaf- and litter-feeding invertebrates as sources of animal protein for the Amazonian Amerindians. AB - At least 32 Amerindian groups in the Amazon basin use terrestrial invertebrates as food. Leaf- and litter-consuming invertebrates provide the more important, underestimated food sources for many Amerindian groups. Further, litter-consuming earthworms are also an important food resource for the Ye'Kuana (also known as Makiritare) in the Alto Orinoco (Amazonas, Venezuela). By selecting these small invertebrates the Amerindians are choosing their animal food from those food webs in the rainforest which have the highest energy flow and which constitute the greatest renewable stock of readily available nutrients. Here we show that the consumption of leaf- and litter-feeding invertebrates as a means of recovering protein, fat and vitamins by the forest-living peoples offers a new perspective for the development of sustainable animal food production within the paradigm of biodiversity maintenance. PMID- 11413640 TI - Experimental evidence for innate predator recognition in the Seychelles warbler. AB - Nest predation is a major determinant of fitness in birds and costly nest defence behaviours have evolved in order to reduce nest predation. Some avian studies have suggested that predator recognition is innate whereas others have stressed the importance of learning. However, none of these studies controlled for the genetic origin of the populations investigated and the effect of unfamiliarity with the predator. Here we determined whether experience with a nest predator is a prerequisite for nest defence by comparing predator recognition responses between two isolated but genetically similar Seychelles warbler (Acrocephalus sechellensis) populations, only one of which had experience of the egg predating Seychelles fody (Foudia sechellarum). Individuals in the predator-free population significantly reduced nest guarding compared to individuals in the population with the predator, which indicates that this behaviour was adjusted to the presence of nest predators. However, recognition responses (measured as both alarm call and attack rates) towards a mounted model of the fody were equally strong in both populations and significantly higher than the responses towards either a mounted familiar non-predator and a mounted, novel, non-predator bird species. Responses did not differ with a warbler's age and experience with the egg predator, indicating that predator recognition is innate. PMID- 11413641 TI - Detection of macro-ecological patterns in South American hummingbirds is affected by spatial scale. AB - Scale is widely recognized as a fundamental conceptual problem in biology, but the question of whether species-richness patterns vary with scale is often ignored in macro-ecological analyses, despite the increasing application of such data in international conservation programmes. We tested for scaling effects in species-richness gradients with spatially scaled data for 241 species of South American hummingbirds (Trochilidae). Analyses revealed that scale matters above and beyond the effect of quadrat area. Species richness was positively correlated with latitude and topographical relief at ten different spatial scales spanning two orders of magnitude (ca. 12,300 to ca. 1,225,000 km2). Surprisingly, when the influence of topography was removed, the conditional variation in species richness explained by latitude fell precipitously to insignificance at coarser spatial scales. The perception of macro-ecological pattern thus depends directly upon the scale of analysis. Although our results suggest there is no single correct scale for macro-ecological analyses, the averaging effect of quadrat sampling at coarser geographical scales obscures the fine structure of species richness gradients and localized richness peaks, decreasing the power of statistical tests to discriminate the causal agents of regional richness gradients. Ideally, the scale of analysis should be varied systematically to provide the optimal resolution of macro-ecological pattern. PMID- 11413642 TI - Testing macro-evolutionary models using incomplete molecular phylogenies. AB - Phylogenies reconstructed from gene sequences can be used to investigate the tempo and mode of species diversification. Here we develop and use new statistical methods to infer past patterns of speciation and extinction from molecular phylogenies. Specifically, we test the null hypothesis that per-lineage speciation and extinction rates have remained constant through time. Rejection of this hypothesis may provide evidence for evolutionary events such as adaptive radiations or key adaptations. In contrast to previous approaches, our methods are robust to incomplete taxon sampling and are conservative with respect to extinction. Using simulation we investigate, first, the adverse effects of failing to take incomplete sampling into account and, second, the power and reliability of our tests. When applied to published phylogenies our tests suggest that, in some cases, speciation rates have decreased through time. PMID- 11413643 TI - Genetic population structure indicates sympatric speciation of Lake Malawi pelagic cichlids. AB - Allopatric processes of speciation have routinely been presented to explain the extraordinary radiation of the East African Great Lakes cichlid fish species flocks. The 21 or more species of pelagic cichlids within the Lake Malawi flock appear to have lake-wide distributions that challenge such a concept. Data from six microsatellite DNA loci indicate single, panmictic populations across the lake of three Diplotaxodon species. Levels of variability at these loci suggest that populations have been large and stable. Mitochondrial DNA sequence data (872 bp of control region + 981 bp of the NADH-2) from 90 species, representing all major clades within the Lake Malawi flock, indicate reciprocal monophyly of the pelagic clade. We suggest that these data support a hypothesis that speciation in sympatry is more plausible (and widespread) within the cichlid species flocks than previously thought. PMID- 11413644 TI - Robustness as an evolutionary principle. AB - We suggest simulating evolution of complex organisms using a model constrained solely by the requirement of robustness in its expression patterns. This scenario is illustrated by evolving discrete logical networks with epigenetic properties. Evidence for dynamical features in the evolved networks is found that can be related to biological observables. PMID- 11413645 TI - Nest predation increases with parental activity: separating nest site and parental activity effects. AB - Alexander Skutch hypothesized that increased parental activity can increase the risk of nest predation. We tested this hypothesis using ten open-nesting bird species in Arizona, USA. Parental activity was greater during the nestling than incubation stage because parents visited the nest frequently to feed their young during the nestling stage. However, nest predation did not generally increase with parental activity between nesting stages across the ten study species. Previous investigators have found similar results. We tested whether nest site effects might yield higher predation during incubation because the most obvious sites are depredated most rapidly. We conducted experiments using nest sites from the previous year to remove parental activity. Our results showed that nest sites have highly repeatable effects on nest predation risk; poor nest sites incurred rapid predation and caused predation rates to be greater during the incubation than nestling stage. This pattern also was exhibited in a bird species with similar (i.e. controlled) parental activity between nesting stages. Once nest site effects are taken into account, nest predation shows a strong proximate increase with parental activity during the nestling stage within and across species. Parental activity and nest sites exert antagonistic influences on current estimates of nest predation between nesting stages and both must be considered in order to understand current patterns of nest predation, which is an important source of natural selection. PMID- 11413646 TI - Day-flying butterflies remain day-flying in a Polynesian, bat-free habitat. AB - To test the theory that insectivorous bats have selected for diurnality in earless butterflies I compared the nocturnal flight patterns of three species of nymphalid butterflies on the bat-free Pacific island of Moorea with those of three nymphalids in the bat-inhabited habitat of Queensland, Australia. Nocturnal flight, measured as the ratio of deep night (1 h following sunset to 1 h preceding sunrise) to twilight night (1 h before sunset to 30 min after sunrise) activity did not differ significantly between the two locations, nor did the percentage of individuals active and I conclude that living in a bat-released habitat has not produced nocturnal flight in these insects. This result is surprising considering the potential advantages of escaping diurnally active predators and suggests that physiological adaptations (e.g. thermoregulation and/or vision) currently constrain these insects to diurnal flight. Since taxonomic records suggest that gene flow does not exist with bat-exposed conspecifics, I suggest that insufficient time has elapsed since these species migrated to Moorea to have resulted in major phenotypic changes such as diel flight preferences. PMID- 11413647 TI - Accurate route demonstration by experienced homing pigeons does not improve subsequent homing performance in naive conspecifics. AB - We describe an experiment that uses the grouping tendencies and navigational abilities of the homing pigeon (Columba livia) to investigate the possibility of socially mediated information transfer in a field setting. By varying the composition of paired-release types, we allowed some naive birds to receive an accurate demonstration of the home route whilst others were paired with similarly naive conspecifics. After this 'paired phase', we predicted that if any learning of spatial information occurred then naive members of the former pairs would outperform their untutored conspecifics when re-released individually during the subsequent 'single phase' of the experiment. This prediction was not confirmed. Neither homing speed nor initial orientation was superior in individually released tutored versus untutored birds, despite the fact that both performance measures were better in the earlier 'paired phase' with experienced demonstrators. Our results suggest that although naive homing pigeons clearly interact with their experienced partners, they are unable to transfer any individually useful spatial information to subsequent homing flights. PMID- 11413648 TI - The developmental emergence of coupled activity as cooperative aggregation in rat pups. AB - Rat pups (Rattus norvegicus) are born blind and deaf yet manage to wriggle about in a huddle, dynamically adjusting their positions and thereby displaying thermoregulation and energy conservation at the level of the group. As pups develop, their activity and mobility outpace the development of their visual and auditory systems making it increasingly difficult to aggregate and maintain aggregation while still blind and deaf. The developmental emergence of coupled activity may be one mechanism that facilitates aggregation. Our previous research has shown that the activity of a seven-day-old pup is independent of the activity of the litter mates it contacts. However, we hypothesized that, by day 10, more active and mobile pups will exhibit coupled activity, becoming increasingly quiescent when in contact with other behaviourally quiescent pups. In order to test this hypothesis, we used individual-based modelling. Because the structure of the model was complex, we used a Darwinian algorithm for evolving a model that behaved like ten-day-old pups aggregating in an arena. Sensitivity to quiescent individuals was manifested in some litters by the transitory spreading of quiescence across aggregates of both real and virtual pups (a contagion effect). As pups develop, individual behaviour becomes increasingly contingent on the behaviour of others revealing what may be a basic component in the development of cooperative behaviour. PMID- 11413649 TI - Development of infant baboons' responses to graded bark variants. AB - We studied the development of infant baboons' (Papio cynocephalus ursinus) responses to conspecific 'barks' in a free-ranging population in the Okavango Delta, Botswana. These barks grade from tonal, harmonically rich calls into calls with a more noisy, harsh structure. Typically, tonal variants are given when the signaller is at risk of losing contact with the group or a particular individual ('contact barks'), whereas harsh variants are given in response to predators ('alarm barks'). We conducted focal observations and playback experiments in which we presented variants of barks recorded from resident adult females. By six months of age, infants reliably discriminated between typical alarm and contact barks and they responded more strongly to intermediate alarm calls than to typical contact barks. Infants of six months and older also recognized their mothers by voice. The ability to discriminate between different call variants developed with increasing age. At two and a half months of age, infants failed to respond at all, whereas at four months they responded irrespective of the call type that was presented. At six months, infants showed adult-like responses by responding strongly to alarm barks but ignoring contact barks. We concluded that infants gradually learn to attach the appropriate meaning to alarm and contact barks. PMID- 11413650 TI - Life in the extreme environment at a hydrothermal vent: haemoglobin in a deep-sea copepod. AB - This is the first study, to my knowledge, quantifying the respiratory pigment haemoglobin discovered in a deep-sea copepod. Haemoglobin in copepods has previously been documented in only one other species from the deep water of an Italian lake. Specimens of the siphonostomatoid Scotoecetes introrsus Humes were collected during submersible dives at 2500 m depth near a hydrothermal vent at the East Pacific Rise (9 degrees N). The haemoglobin content in the copepods' haemolymph was 4.3 +/- 0.6 micrograms per individual female (n = 6) and 1.8 +/- 0.1 micrograms per individual male (n = 6). Weight-specific concentrations of haemoglobin were identical for females and males (0.25 +/- 0.04 and 0.26 +/- 0.02 microgram per microgram dry weight, respectively). These haemoglobin concentrations are higher than those found in other small crustaceans. Activity of the electron transport system indicated that the respiration rates in S. introrsus (13.7 +/- 7.7 microliters O2 per milligram dry weight per hour) were similar to those in the shallow-water copepod Acartia tonsa (9.1 +/- 1.3 microliters O2 per milligram dry weight per hour). It was concluded that the possession of highly concentrated haemoglobin allows S. introrsus to colonize a geologically young, thermally active site such as the vicinity of a hydrothermal vent, despite the prevailing oxygen depletion. PMID- 11413651 TI - Induced pigmentation in zooplankton: a trade-off between threats from predation and ultraviolet radiation. AB - Ultraviolet (UV) radiation is harmful to all life, and the ongoing depletion of the ozone layer is likely to affect interactions among both terrestrial and aquatic organisms. Some organisms have evolved adaptations to reduce radiation damage, such as the various types of protective pigmentation of freshwater zooplankton. However, strong pigmentation also increases vulnerability to visually hunting predators. Hence, where both UV radiation and predation are intense, zooplankton may be sandwiched between conflicting selective pressures: to be pigmented and to be transparent at the same time. Here, I show that the level of pigmentation in copepods is up to ten times higher in lakes without predatory fishes than where fishes are present. Moreover, animals from the same population exposed to either UV light or predator scent showed a 10% difference in pigmentation after only four days, suggesting that pigmentation is an inducible trait. Hence, individual copepods are not passive victims of selective predation or radiation damage, but adjust the level of pigmentation according to the prevailing threat. The ability to adjust pigmentation level rapidly may be especially useful in situations where risk assessment is difficult due to strong seasonal and spatial variation in risk variables, such as in Arctic regions. With progressive thinning of the ozone layer, the ability of some but not other organisms to adjust protection against UV radiation may lead to counter intuitive, large-scale alterations in freshwater food webs. PMID- 11413652 TI - Reduced sperm counts in guppies (Poecilia reticulata) following exposure to low levels of tributyltin and bisphenol A. AB - There is increasing evidence that normal male reproductive function can be disrupted by exposure to pollutants in the environment that can exogenously mimic, antagonize or block sex-hormone function. One possible consequence of exposure to these xenobiotics is disruption to spermatogenesis, but results thus far provide only indirect and inconsistent evidence. In this study we exposed adult male guppies (Poeciliidae: Teleostei) to environmentally relevant levels of the common xenobiotics tributyltin (11.2-22.3 ngl-1) and bisphenol A (274-549 micrograms l-1) in experimental aquaria. After 21 days of exposure, we found significant declines (by 40-75%) in total sperm counts for male fishes exposed to tributyltin and bisphenol A compared with controls. This short-term decline in sperm count is unlikely to be the result of endocrine-mediated alteration of the germ line, and we found no change in testis size or sperm lengths between treatments. However, Sertoli cells, which facilitate the transport of maturing sperm into the testicular deferent duct (where they are stored prior to ejaculation), are directly sensitive to xenobiotic action and it is therefore possible that spermatogenesis was inhibited through in vivo interference with normal Sertoli-cell function. PMID- 11413653 TI - Testosterone, ticks and travels: a test of the immunocompetence-handicap hypothesis in free-ranging male sand lizards. AB - The immunocompetence-handicap hypothesis suggests that androgen-dependent male characters constitute honest signals of mate and/or rival quality because of the imposed costs through immune suppression associated with elevated testosterone levels. We demonstrate in a field experiment that male sand lizards (Lacerta agilis) exposed to elevated testosterone suffered from increased mass loss and tick load compared to control males. Although the first of these two results could be due to an elevated basal metabolic rate from increased plasma testosterone levels, the increased parasite load was statistically independent of the loss in body condition and is likely to be due to compromised immune function. Testosterone-treated males showed greater mobility than control males, and greater mobility resulted in higher mating success. Our experiment thus lends support to the immunocompetence-handicap hypothesis, suggesting that male testosterone levels have been moderated by balancing selection for reproductive success and sustained immune function. PMID- 11413655 TI - You don't know until you need one. PMID- 11413654 TI - Species concepts and malaria parasites: detecting a cryptic species of Plasmodium. AB - Species of malaria parasite (phylum Apicomplexa: genus Plasmodium) have traditionally been described using the similarity species concept (based primarily on differences in morphological or life-history characteristics). The biological species concept (reproductive isolation) and phylogenetic species concept (based on monophyly) have not been used before in defining species of Plasmodium. Plasmodium azurophilum, described from Anolis lizards in the eastern Caribbean, is actually a two-species cryptic complex. The parasites were studied from eight islands, from Puerto Rico in the north to Grenada in the south. Morphology of the two species is very similar (differences are indistinguishable to the eye), but one infects only erythrocytes and the other only white blood cells. Molecular data for the cytochrome b gene reveal that the two forms are reproductively isolated; distinct haplotypes are present on each island and are never shared between the erythrocyte-infecting and leucocyte-infecting species. Each forms a monophyletic lineage indicating that they diverged before becoming established in the anoles of the eastern Caribbean. This comparison of the similarity, biological and phylogenetic species concepts for malaria parasites reveals the limited value of using only similarity measures in defining protozoan species. PMID- 11413656 TI - Why neuroscience nursing. PMID- 11413657 TI - Description and use of the neuroscience nursing self-efficacy scale. AB - An instrument for assessing nurses' perceived self-efficacy in implementing a variety of neuroscience nursing tasks was developed. Self-efficacy theory served as the guiding framework. From 1988 to 1998, the instrument was used to assess changes in the perceived self-efficacy of 54 nurses who attended a neuroscience nurse-internship program. Self-efficacy was assessed during clinical orientation, prior to the beginning of the program, and at the end of the program. The results showed that the nurses' confidence in performing a variety of neuroscience nursing skills increased during the 6- to 10-month program. The instrument was also useful in helping program directors identify areas in which nurses could benefit most from the program and refine the program to meet the educational needs of the nurses. PMID- 11413658 TI - Remotivation therapy and Huntington's disease. AB - Huntington's disease (HD) is a genetic, autosomal dominant, neurodegenerative disorder for which there is no known cure. Because remotivation therapy (RmT) has been effective in other neurological conditions and because of the lack of current efficacious treatments for HD, RmT may be a beneficial therapy for persons suffering from the disease. By cultivating a more fertile environment, RmT leads to increased self-awareness, self-esteem, and an improved quality of life, even in late-stage HD. Two recent studies using animal models suggest that exposure of transgenic HD mice to a stimulating, enriched environment helped to prevent the loss of cerebral volume and delay the onset of motor disorders. Six case studies are presented that demonstrate improved physical, mental, and social functioning in persons with HD when a more stimulating environment is provided. PMID- 11413659 TI - Being a brain tumor patient: a descriptive study of patients' experiences. AB - The purpose of this study was to describe the experience of being a patient with a brain tumor in the neurosurgery clinic of a university hospital. Eight brain tumor patients volunteered to participate. The data were collected by interviewing the participants on the day preceding brain surgery and 3-7 days postoperatively. The interviews were analyzed by inductive content analysis. Preoperatively, some patients had a fearless and calm attitude towards their illness, while others were fearful and depressed. Postoperatively, the patients' body images changed, and they were concerned about the future. They perceived their care as matter-of-fact and friendly both before and after the surgery, and they thought their basic needs were met. Some patients would have wanted more psychological support, especially after the surgery, while others found the psychological support adequate. The patients were willing to participate in decision making about their care, and they trusted the professional skill and competence of the nurses. Suggested improvements in care were to minimize the atmosphere of urgency and hurry, appoint a primary nurse for each patient, and give more attention to after-care. PMID- 11413660 TI - Gastric versus duodenal feeding in patients with neurological disease: a pilot study. AB - Both gastric and duodenal feeding tubes are used to provide enteral nutrition. Most studies comparing the two methods have focused primarily on rates of complications, rather than on nutritional outcomes, and show no difference in complications between the two methods. It is not clear which feeding route provides the best nutritional outcomes. The primary purpose of this randomized clinical pilot study was to compare the percentage of recommended calories and protein received by patients with neurological disease being fed enterally via gastric or duodenal feeding tubes. Secondary aims were to compare the following between groups: physiological effects of feeding, reasons for delay in feeding, volume of feeding residual, number of feeding tubes replaced, cost of feeding, and number and types of complications. A convenience sample of 25 neuro intensive care unit patients was randomly assigned to gastric or duodenal feeding. Enteral feeding was ordered by using a standardized prescription formula and provided by the nursing staff. Serum albumin and prealbumin levels were measured at baseline, day 3, and day 10. Nitrogen balance was measured on day 10. Enteral feeding data were collected daily. No significant differences were found between gastric and duodenal groups in nutritional outcomes, including percentage of recommended calories and protein received, physiological effects of feeding, reasons for delay in feeding, feeding residual, number of feeding tubes replaced, cost of feeding, and number and types of complications. Neither group achieved mean recommended caloric or protein intake during the 10 days of the study. Further research is needed to address how recommended nutrients can be provided enterally in a more timely and complete manner in critically ill NICU patients. PMID- 11413661 TI - Barriers that further disablement: a study of survivors of polio. AB - This exploratory study of polio survivors was undertaken to clarify current level of function, comorbidity, and secondary conditions and to discover the relationships between perceived barriers to health, functional limitation, comorbidity, secondary conditions, and disability in a small convenience sample. Given the underrepresentation of research in the literature explaining the health status, perceived function, and barriers to health in this disabled population, a descriptive study with correlation was conducted. Thirty-one individuals living with the effects of polio were surveyed. Disability was correlated with perceived barriers to health but not with functional limitations, comorbidity, or secondary conditions. Secondary conditions were correlated with the number of years living with the illness. The findings support a model of disablement and allostatic load. Further research is needed to test these models and to provide direction for nurses caring for those aging with the effects of polio. PMID- 11413662 TI - Problems reported by elderly patients with multiple sclerosis. AB - Multiple sclerosis (MS) is one of the most common organic neurological diseases of the central nervous system. Because of improved therapies, nurses are confronted with elderly MS patients, but little is known about the specific problems of these patients. This survey analyzed problems in elderly MS patients. Fifty-three MS patients (44 female, 9 male; average age 73 years, average course of MS 25.3 years) from the Berlin Section of the German Multiple Sclerosis Association were evaluated by using a standardized questionnaire, considering social situation, daily problems, disease course, and disabilities, and by using the expanded disability status scale (EDSS). Elderly MS patients reported impaired mobility and inability to use public transportation; about 96% presented EDSS scores above 6.0. Nearly 50% complained about spasticity and pain due to spasticity. More than 70% suffered from bladder dysfunction. Problems with sleep and fatigue were present in less than 20%, but interrupted sleep was common. Selfcare impairments were reported by 50%-75% of the patients, and most of them required professional help. Depressive moods and thoughts about committing suicide were mentioned by more than 30% of the patients. Elderly MS patients experience physical and psychosocial impairments. Healthcare professionals should consider increasing independence and avoiding nursing home admissions in the management of elderly MS patients. PMID- 11413674 TI - Bibliography. Current world literature. Respiratory system. PMID- 11413675 TI - [Effects of new antineoplastic agents (dimethoxybenflurone and Oracin) on the cardiovascular system and other parameters in the rabbit in vivo]. AB - Anthracycline derivatives belong among the most effective antineoplastic drugs but their therapeutic use is limited by their side effects--a dose-related cardiotoxicity. The influence of repeated i.v. administration (once weekly, max. 10 weeks) of new antineoplastic agents--dimethoxybenfluron (DMB) (3,9 dimethoxybenfluron hydrochloride, C23H24O4NCl, M.w. 413.9, Institute of Experimental Biopharmaceutics, Czech Academy of Sciences, Hradec Kralove, Czech Republic; 12 or 24 mg base/kg) and Oracin (6-[2-(2-hydroxyethyl)aminoethyl]-5,11 dioxo-5,6-dihydro-11H- indeno[1,2c]isoquinoline hydrochloride), C20H19N2O3Cl, M.w. 370.84, Research Institute for Pharmacy and Biochemistry, Prague, Czech Republic; 5 or 10 mg/kg) on cardiovascular, biochemical, haematological and histological parameters were studied in rabbits in vivo. Data obtained in these groups were compared with the group with experimentally induced cardiomyopathy (daunorubicin 50 mg/m2 i.v.) and with the control group (saline 1 ml/kg). Only mild and mostly no significant changes of the cardiovascular parameters (DMB 12 group: PEP:LVET ratio--0.408-0.502, LV dP/dtmax.--1337.0 kPa/s; DMB 24 group: PEP:LVET ratio--0.407-0.433, LV dP/dtmax.--1438.2 kPa/s), biochemical parameters (decrease in natrium, ALP and increase in glucose, GPX and GSH levels) and haematological parameters (increase in erythrocytes and decrease in leukocytes after the larger dose of the drug) were found in the dimethoxybenfluron groups. Repeated administration of the lower dose of Oracin induced only mild and mostly no significant changes of parameters (PEP:LVET ratio--0.393-0.475, LV dP/dtmax.- 1092.4 kPa/s) in comparison with the control group. Though significant in some intervals, only a mild oscillation of the PEP:LVET ratio (0.368-0.446), decrease in LV dP/dtmax. (991.2 kPa/s) and--in comparison with control group- significantly higher blood pressure and lower heart rate were found after the higher dose of Oracin. In the most of haematological and biochemical parameters (with the exception of chlorides, protein and albumin levels) no significant changes were present. Histological examination of the heart revealed normal structure of the myocardium including minute changes of myocardium following administration of antineoplastic agents in all groups. Administration of new antineoplastic agents induced mostly mild changes of the followed-up parameters (PEP:LVET ratio, LV dP/dtmax., heart rate, levels of cardiac troponin T, survival of animals, haematological and biochemical parameters); the values of parameters were mostly significantly different from those in rabbits with daunorubicin induced cardiomyopathy. On the basis of our results it is possible to conclude that the administration of dimethoxybenflurone and Oracin did not induce signs of cardiotoxicity in rabbits in vivo. This observation is considered to be important from the viewpoint of possible further clinical use of these new antineoplastic agents. PMID- 11413676 TI - [Renal excretion of methotrexate in an in vivo model in minipigs]. AB - The goal of the study was to evaluate a minipig (i.e. porcine) model as the human like model for preclinical evaluation of mechanisms involved in the renal excretion of high-dose methotrexate (HDMTX). Methotrexate is in man renaly excreted in combination of glomerular filtration and active tubular drug transport (in a proximal tubule). After intravenous MTX administration, more than 95% of the amount of delivered dose was detected in urine in the form of intact MTX. To compare glomerular filtration and other ways of renal MTX excretion, the ratio between MTX clearance and clearance of inuline (which evaluate the rate of glomerular filtration only) was calculated and analyzed. Renal clearance of MTX was higher than that of inuline (Cl/Cl = 1.50 (0.095 ml/min.kg). The results showed a significant correlation between Cl and pH of urine (r = 0.525, r = 0.7243, p < 0.001, figure 1). Similar correlations were found when comparing the results of Cl and glomerular filtration (r = 0.8589, r = 0.8939, p < 0.00001) (figure 2). Significant relationship was also evident between Cl and urine pH and GF together (simultaneously) (r = 0.8677). The renal clearance of MTX varied from 1.36 ml/min.kg (measured at pH 6.0) to 3.2 ml/min.kg (measured at pH 7.0). Finally, the results indicate a significant relationship between the renal and extrarenal clearance MTX (r = 0.7227, p < 0.0001). PMID- 11413677 TI - [Basic principles of physics in optical imaging of the human eye]. AB - Creation of a real picture of the observed object on the retina by optical system of the eye is the primary physical condition of the visual perception. Absorption of the light energy initiates the sequence of further physicochemical, biochemical and psychophysiological events partially taking place already in the retina but mostly in the higher cerebral and spinal centres. So the visual perception and its final evaluation is the result of a complicated complex of processes where the optical imaging itself plays the role of the starting factor. In the publication particularly the physical substance of the optical imaging on refraction surfaces between media with different refraction index has been accentuated. The laws of reflection and refraction and their origin are derived in the terms of the physical (Maxwell equations) and geometrical (Fermat principle, principle of the mutual independence of light beams) optics. In the adequate extent the development and structure of the eye and the whole visual complex, the bases of the quantification of the visual perception and contemporary opinions on mechanisms of the colour vision (trichromatic theory) have been described. The publication is completed by a mathematical appendix explaining some relations usually absent in the biomedical literature. PMID- 11413678 TI - Trends in pregnancy rates for the United States, 1976-97: an update. AB - OBJECTIVES: This report presents detailed pregnancy rates for 1996 and 1997 to update a recently published comprehensive report on pregnancies and pregnancy rates for U.S. women. METHODS: Tabular and graphic data on pregnancy rates by age, race, and Hispanic origin, and by marital status are presented and described. RESULTS: In 1997 an estimated 6.19 million pregnancies resulted in 3.88 million live births, 1.33 million induced abortions, and 0.98 million fetal losses. The 1997 pregnancy rate of 103.7 pregnancies per 1,000 women aged 15-44 years is the lowest recorded since 1976 (102.7), the first year for which a consistent series of national pregnancy rates is available. The 1997 rate was 10 percent lower than the peak rate in 1990 (115.6). The teenage pregnancy rate dropped steadily through 1997, falling to a record low of 94.3 pregnancies per 1,000 teenagers 15-19 years, 19 percent below the 1990 level (116.3). Rates for younger teenagers declined more than for older teenagers. PMID- 11413679 TI - [Nontargeted mutagenesis of plasmid pSP189 induced by nickel acetate and its relation with DNA damage in Vero cells]. AB - OBJECTIVE: To detect the nontargeted mutagenesis and DNA damage in Vero cells induced by nickel acetate and to analyze their association, to provide clues to carcinogenic mechanism of nickel. METHOD: Vero cells were treated with nickel acetate for 2.5 hours and cultivated in routine condition for 24 hours, and transfected with wild plasmid pSP189. Plasmid replicated in mammalian cells was collected and was transferred into E. coli MBM7070. The mutant was selected with special culture media. DNA damage in Vero cells was detected with modified single cell gel electrophoresis. RESULTS: Nontargeted mutagenesis occurred in the cells pretreated with 250 mumol and 1,000 mumol nickel acetate, with mutagenesis frequencies of 9.46 x 10(-4) and 15.01 x 10(-4), 4.41 and 6.98 folds more as compared with the control group, respectively. Frequencies of DNA damage, DNA chain breaks, DNA-protein cross-links and DNA-DNA cross-links in all pretreated cells were significantly higher than those in the control group, but without dose effect relationship. There was dose leap for mutagenesis frequencies and DNA-DNA cross-links. CONCLUSION: Nickel acetate could induce nontargeted mutagenesis in the system of shuttle plasmid vectors, which could be related to DNA-DNA cross links. PMID- 11413680 TI - [Effects of tobacco smoke on the expression of ICAM-1 in human umbilical vein endothelial cells]. AB - OBJECTIVE: To study the effect of tobacco smoke on the expression of intercellular adhesion molecule (ICAM)-1 in endothelial cell. METHODS: Macrophages were challenged in vitro with cigarette smog, and the supernatant was collected at 3 h, 6 h, 12 h and 24 h after tobacco treatment. The tumor necrosis factor-alpha (TNF-alpha) was measured by the ELISA method. Then the human umbilical vein endothelial cells (HUVEC) were incubated with the supernatant from the macrophages stimulated by cigarette smoke allergen. ICAM-1 in the supernatant was detected by the ELISA method. RESULTS: TNF-alpha was increased with the longer incubation time. A significant difference of TNF-alpha(t = -4.301, P < 0.01) was observed between the tobacco treated group (4.967 +/- 0.802 pg/ml) and the control group (3.181 +/- 0.214 pg/ml). Soluble ICAM-1 in HUVEC in the tobacco treated group (mean 0.068 4 ng/ml) increased 49% than the controls (mean 0.045 9 ng/ml). CONCLUSION: The results suggested that cigarette smog enhances the expression of TNF-alpha in the antigen-presenting cell(APC)-macrophage, and the latter up-regulates the secretion of the membrane glycoprotein sICAM-1 in HUVEC which can assist leukocyte immigration. PMID- 11413681 TI - [Inhibition of voltage-activated outward delayed rectifier potassium channel currents in dorsal root ganglion neurons of rats by lead]. AB - OBJECTIVE: To study the effect of lead on voltage-activated outward delayed rectifier potassium channel currents in acute isolated dorsal root ganglion (DRG) neurons of adult rats. METHODS: Patch clamp technique was employed to record the whole cell of DRG in adult rats. RESULTS: In the experiments, delayed rectifier (IK) potassium currents channel was recorded and activation was voltage dependent. The threshold concentration of lead (Pb2+) for IK was 1 mumol/L, which reduced potassium current by (8.6 +/- 0.8)%, 4 mumol/L of Pb2+ reduced the current by (38.6 +/- 6.2)%, and 8 mumol/L of Pb2+ reduced the current by (63.0 +/ 5.1)%. The action of lead was rapid and its effect would be reversed rapidly to various extent by washing. CONCLUSION: Higher outward delayed rectifier potassium channel currents, which could be reversed. PMID- 11413682 TI - [Detection of gangliosides in human milk with a high performance thin layer chromatography (HPTLC)]. AB - OBJECTIVES: To determine the ganglioside composition in human milk during lactation, and to evaluate the biological significance. METHODS: The gangliosides extracted from 68 milk samples were examined with a high performance thin layer chromatography (HPTLC). RESULTS: The concentration of total lipid bound sialic acid was 7.8-10.4 microliters/ml. Four gangliosides were found and designated as GM3, GD3, GX1 and GX2, respectively. GX1 and GX2, which had not been described previously, were tentatively assumed to be gangliosides of the c-pathway because they did not react to the GA1 antibody after sialidase treatment. GD3 and GM3 were the major gangliosides in human milk (accounted for 50%-65%). CM3 decreased sharply at eight days postpartum. Both GM3 and GD3 changed significantly during lactation (GM3, r = 0.65, P < 0.01; GD3, r = -0.58, P < 0.01). CONCLUSIONS: Four gangliosides, GM3, GD3, GX1 and GX2 were found in human milk. Both GM3 and GD3 changed significantly during lactation. PMID- 11413683 TI - [Effects of taurine on brain development and learning ability in filial mice fed with high iodine]. AB - OBJECTIVE: To observe the effects of taurine on brain development and learning ability in filial mice fed with high iodine. METHODS: Mice were divided into three groups, the first one fed with adequate dose of iodine, the second with high-dose iodine, and the third with high-dose iodine plus 0.6% taurine. All the male and female mice were mated three months after treatment. Relevant biochemical indicators in the brain tissues and their memory and learning ability were measured for their offsprings with 30 days old in the three groups. RESULTS: Brain contents of protein and DNA reduced in the high-dose iodine group, and there was no significant difference in them between groups with taurine and adequate-dose iodine 30 days after their birth. Activity of acetylcholine esterase (ACHE) increased and content of acetylcholine decreased in the hippocampus of filial mice at age of 30 days with high-dose iodine. Content of acetylcholine increased in the filial mice with high-dose iodine plus taurine than that with adequate-dose iodine, but there was no statistically significant difference in ACHE between them. CONCLUSION: Taurine could antagonize disturbance of brain development in filial mice caused by high-dose iodine. PMID- 11413684 TI - [Analysis of HIV-1 seropositive individual infected with different subtypes]. AB - OBJECTIVE: To investigate whether there existed any individuals infected with different subtypes of human immunodeficiency virus type 1 (HIV-1) and to analyze their source of infection. METHODS: Membrane protein gene (env) of HIV-1 in uncultured single peripheral blood monocytes (PBMCs) from HIV-1 seropositive individuals found in Shenzhen was amplified by nested polymerase chain reaction (PCR). The C2-V3 region and its adjacent region (360 bp) of them were sequenced and analyzed, and their divergence rate and systematic phylogenetic tree (cladogram) were calculated. RESULTS: There existed sequences of B and C or B and E strains of HIV-1 subtype in three infected individuals in Shenzhen. Gene divergence rates between different subtypes and corresponding international subtype reference strains Bcon, Ccon and Econ were 5.07%-7.26%, 3.23% and 5.38% 5.99%, respectively, and those between different subtypes strains in the same individual were 23.52%, 20.69% and 28.44%, respectively. GPGR sequence of typical Europe and America subtype B was found in strain sz46-1 from prostitute with intravenous drug abuse, based on their tetra-peptide sequence characteristics in the crown of gp 120 V3 loop, strain sz8-2 was GPGH and other strains were GPGQ sequence characterized by Thai B (B') and E subtypes. Furthermore, phylogenetic tree analysis showed that strains sz1-2, sz8-1 and sz46-1 were clustered with international reference sequence Bcon, strain sz1-1 clustered with Econ, and strains sz1-1 and sz8-2 clustered with Ccon. CONCLUSION: These findings suggested that an individual could be simultaneously infected with different HIV-1 subtypes, which could probably be caused by his or her own multiple high-risk behavior. PMID- 11413685 TI - [Effects of comprehensive intervention on prognosis of isolated systolic hypertension in the elderly]. AB - OBJECTIVE: To study the effects of comprehensive intervention on prognosis of isolated systolic hypertension in the elderly. METHODS: A 12-years prospective and controlled study was conducted with 207 elderly patients of isolated systolic hypertension enrolled. According to the study project and procedure, 99 patients were allocated to the intervention group and 108 patients in the control group. All the events of cardiovascular diseases, cerebrovascular disorders, nephrovascular diseases and death were recorded and analyzed. RESULTS: A 12-year consecutive follow-up showed that the incidence and mortality of cardiovascular diseases, cerebrovascular disorders, nephrovascular diseases and their ages at death were 41.49%, 26.60% and 75.6 years, respectively, in the intervention group, and 77.45%, 53.92% and 70.2 years, respectively, in the control group, with statistically significant difference. Survival rate in the intervention group was higher than that in the control group. CONCLUSION: Comprehensive intervention for a long time could remarkably improve their clinical prognosis of the elderly patients with isolated systolic hypertension. PMID- 11413686 TI - [Study on the effects of DNA and natural killer cell damage induced by microcystins LR]. AB - OBJECTIVE: Microcystin is a group of algae toxin which causes liver cancer. In this paper we tried to study the carcinogenic and tumor promotion mechanism of Microcystin LR (MCLR). METHODS: The single cell microgel electrophoresis assay (MGE) and 3H-TdR releasing assay were used to study the effects of on DNA damage and natural killer cell (NK cell) activity. RESULTS: The different doses of MCLR caused DNA damage in rat lymphocytes compared to 0.85% NaCl. However, no time dependent relationship and dose dependent relationship between MCLR and DNA migration were observed, and the DNA damage was recovered rapidly after MCLR treatment. At the same time MCLR reduced NK cells activity in mice. CONCLUSION: The above results provide some evidence on the mechanisms of carcinogenicity and promotional effects of MCLR at cellular and molecular levels. PMID- 11413687 TI - [Protective effects of organic acids on human vascular endothelial cells]. AB - OBJECTIVES: In order to examine if organic acids (OA), a kind of natural food components, will prevent from atherosclerosis formation, the effects of four OA, namely chlorogenic acid (CHA), ascorbic acid (AA), citric acid (CA), and malic acid (MA) on human vascular endothelial cells were studied. METHODS: Primary human umbilical vein endothelial cells (EC) were cultured, and the changes of cell morphology, cell growth, lactate dehydrogenase (LDH) released from cells were measured when EC was incubated with oxidized low density lipoprotein (oxLDL) in the presence or not of four OA added at three different conditions, i.e. before 4 hours oxLDL added, at the same time as oxLDL added, and after 3 hours oxLDL added, as well as at the indicated doses (10, 20 or 40 mg/L). RESULTS: The EC survival rate of the oxLDL control group was lower (P < 0.01) and LDH release rate was higher (P < 0.01) than those of normal control group. CHA, AA, CA and MA protected EC from damage caused by oxLDL when they were added before and/or at the same time of oxLDL, but the same beneficial effects of these four OA added after oxLDL were not observed. The EC survival rates of CHA + oxLDL and AA + oxLDL groups in three doses were higher (P < 0.05) and the LDH release rates were lower (P < 0.05) in a dose-effect manner when compared with those of oxLDL control group, respectively. Similar results were found in the CA + oxLDL and MA + oxLDL of the high dose groups. CONCLUSION: Four tested OA could effectively prevent EC from damage induced by oxLDL. CHA and AA had the strongest effects, the next was CA, followed by MA. PMID- 11413688 TI - [Effects of exposure to low-level benzene and its analogues on reproductive hormone secretion in female workers]. AB - OBJECTIVE: To study the effects of exposure to low level of mixed benzene on reproductive hormone secretion during menstrual cycle in female workers. METHODS: Concentrations of benzene, toluene and xylene in the expiratory air of 50 exposed female workers from their breathing zone were determined with gas chromatography. Their menstrual cycles of 50 exposed workers and 35 internal control and 35 external control workers were studied prospectively. Urine pregnandiol-3 glucuronide (PdG), follicle stimulating hormone (FSH) and estrone conjugate (E1C) were measured by enzyme immunoassay. RESULTS: Mixed benzene existed mainly in the form of low-level benzene in the air of workplaces, with a detection rate of 29.10% at an average level of 8.88 (0.90-876.47) mg/m3, and 21% of measurements exceeded the national maximum allowable concentration with the highest one as 20.91 folds high as that of the national hygienic standard. Length of luteal phase in the exposed group (13.7 +/- 1.5) days was significantly shorter than that in the internal and external control groups (14.5 +/- 1.2) days and (15.2 +/ 1.1) days (P < 0.05 or P < 0.01). Urine level of E1C before ovulation, that of FSH at early follicular phase and that of PdG in luteal phase after ovulation in the exposed group were significantly lower than those in the internal control group (P < 0.05 or P < 0.01). CONCLUSION: Exposure to low level of mixed benzene in workspaces could interrupt the function of hypothalamic-pituitary-ovarian axis and affect their normal levels of FSH, PdG and E1C. PMID- 11413689 TI - [Study on frequencies of aneuploidy in mouse oocyte and female pronucleus of one cell zygote induced by benzene]. AB - OBJECTIVE: To study the effect of benzene on aneupoidy of oocytes and female pronucleus of one cell zygotes. METHOD: The NIH adult female mice were treated with varies doses of benzene (942, 1,881 and 3,762 mg/kg respectively) via single gavage and (706, 1,922 and 4,864 mg/m3, respectively) multiple inhalation. After gavage, the mice were pair-matched (1:1) with males overnight. The ovulated oocytes and the one cell zygotes were collected for cytogenetic analysis and the frequencies of aneuploidy were detected. RESULTS: The frequencies of aneuploid in M II oocytes significantly increased in the three groups treated with inhalation (7.06%, 7.50% and 7.76%, respectively, control group 0.00%, P < 0.05) of benzene, as compared with those in the control group (1.30%), with a dose-dependent response. Meiotic delay of M I oocytes was observed, with the frequency of M I oocytes of 1.16%, 3.61% and 5.75%, respectively, P < 0.05, in a dose-dependent response, too. In gavage treatment group, increase in frequency of M II oocyte aneuploidy was only observed in the high-dose group, and aneupliody frequency of female pronucleus in one cell zygotes was not induced. CONCLUSION: Exposure to high-dose benzene via gavage or inhalation was able to induce increase in aneuploidy frequency of M II oocytes in mice. PMID- 11413690 TI - [Effects of chemical agents on the prevention of fungal contamination during household wheat storage in Cixian county]. AB - OBJECTIVE: To evaluate the effectiveness of several commercial chemical agents (DMF, AP and Chumeijing) on the prevention of fungal contamination in household storage of wheat in Cixian county. METHODS: The antifungal agents tested include dimethyl fumarate, DMF, aluminium phosphide, AP and Chumeijing (a Chinese product comprised of propionic acid, formic acid, sorbic acid and fumaric acid). The chemicals were applied in the wheat containers in the rural households. The fungal contamination and the inhibitory effects on fungal growth in wheat were measured by fungal isolation, culture, identification and count. RESULTS: The fungal contamination in the household wheat before using fungicides were quite common, with an overall contamination rate of 67.5% to 100%. The application of the three chemical fungicides effectively reduced fungal contamination in household stored wheat, and the effective rate for DMF, AP, AMF + AP and Chumeijing was 91.2%, 85.2%, 100% and 100% respectively, while the rate of fungi growth inhibition was (61.9 +/- 21.6)%, (64.0 +/- 21.6)%, (72.5 +/- 18.0)% and (63.1 +/- 24.3)% respectively. CONCLUSION: DMF, AP and Chumeijing are all effective in the prevention of fungal contamination for household stored wheat in Cixian county. AP is not recommended owing to its higher toxicity, compared with the other chemicals tested. PMID- 11413691 TI - [The event-related potential characteristics of reading disabilities in children under condition of continuous performance test]. AB - OBJECTIVE: To investigate the characteristics of event-related potential (ERP) of reading disabilities (RD) in children under the condition of continuous performance test (CPT) and their possible neurological basis. METHODS: Using ERP technique with CPT conditions, such as semantics, direction and pitch identification, 16 RD boys and their matched normal boys were tested. A comparative study was conducted to analyze these indicators of accuracy, reaction time, false alarm, wave amplitude and latency between the two groups. RESULTS: Among the three kinds of identification tests, RD children had lower accuracy in pitch identification (65.4 +/- 15.9) to (78.5 +/- 12.6) and lower reaction speed (557.0 +/- 97.8) ms to (493.0 +/- 47.8) ms, as compared to the control group (P < 0.05). The false alarm rate was much higher in RD children than that in the control group with unattended-high simulation (1.1 +/- 0.7)% to (0.6 +/- 0.3)%, P < 0.05. Moreover, the P300 amplitude decreased with direction of (20.8 +/- 7.3) to (27.7 +/- 8.3) microV and pitch of (9.1 +/- 4.3) to (14.6 +/- 8.3) microV, P < 0.05. And, the latency delayed in pith of (571 +/- 78) ms to (512 +/- 62) ms, semantic cognition processing negativity Nd of (398 +/- 76) ms to (342 +/- 67) ms, pitch Nd of (373 +/- 56) ms to (327 +/- 53) ms, P < 0.05. CONCLUSIONS: There was defect in selective attention of RD children, which suggested relationship between dysfunction of frontal-basal ganglia circle and RD. The ERP combined with CPT was much better than that with single target stimulation to indicate children's cognitive characteristics. PMID- 11413692 TI - [Relationship between delta-aminolevulinic acid dehydratase gene polymorphism and blood levels of lead and zinc protoporphyrin in children exposed to lead]. AB - OBJECTIVE: To explore the associations of polymorphism of delta-aminolevulinic acid dehydratase (ALAD) gene with blood levels of lead and zinc protoporphyrin (ZPP). METHODS: Blood samples were collected from 370 children exposed to a higher level environmental lead and their blood levels of lead and ZPP, and ALAD genotype were examined. Exposure to lead condition, such as parents' lead exposure and sites of lead source around their residence, and age and sex were matched in ALAD1-1 and ALAD1-2/2-2 subgroups. RESULTS: Blood lead level was higher in the ALAD1-2/2-2 subgroup than that in the ALAD1-1 children [(2.621 +/- 0.561) mumol/L vs (2.360 +/- 0.596) mumol/L, P = 0.04)]. Blood ZPP level was also higher in with ALAD1-2/2-2 subgroup than that in the ALAD1-1 genotype [(13.07 +/- 9.38) mumol/L vs (9.90 +/- 6.30) mumol/L), P = 0.02)]. CONCLUSION: The results suggest that the ALAD1-2/2-2 genotype may influence the body burden and blood toxicity of lead in high lead exposed children. PMID- 11413693 TI - [Trend of changes in mortality of cardiovascular diseases in some areas of Beijing during 1984 to 1998]. AB - OBJECTIVE: To understand the main causes of deaths affecting population health and the trend of mortality of cardiovascular disease (CVD) in Beijing area during 1984 to 1998. METHODS: Monitoring data for cardiovascular diseases in some areas of Beijing during 1984 to 1998 were collected from MONICA project and used to analyze the distribution of disease, mortality caused by cardiovascular diseases and its age-adjusted mortality. Yearly average change in cardiovascular disease specific mortality was calculated with regression analysis. RESULTS: Cardiovascular disease-specific mortality ranked the first place of the causes of deaths in Beijing population during 1984 to 1998, accounting for 46.0% of the total deaths. Cancer ranked the second, accounting for 26.5% of the total. Overall mortality, cardiovascular-specific mortality and cerebro-specific mortality all lowered significantly during the past 15 years, with yearly average reductions of 2.9%, 2.7% and 3.5%, respectively. Mortality of coronary heart disease significantly rose. CONCLUSIONS: Cardiovascular disease was the top cause of death in Beijing population during 1984 to 1998. Mortality of coronary heart disease significantly rose in the past 15 years. It is a pressing matter of the moment to prevent cardiovascular diseases in the population. PMID- 11413694 TI - [Current anesthesia techniques--an attempt at classification]. PMID- 11413695 TI - [Sympathetic control mechanisms during general anesthesia]. AB - For more than 100 years scientists have studied the sympathetic nervous system and its cardiovascular control mechanisms. Muscle sympathetic activity is the most important direct and rapidly responding variable for evaluation of sympathetic neural outflow. Therefore, anesthesiologists have been interested in the effects of anesthetics on muscle sympathetic activity at rest and during cardiovascular challenges. This review summarizes effects of positive pressure ventilation, intravenous and inhalational anesthetics as well as nitrous oxide on sympathetic muscle outflow. The least depression of both resting sympathetic outflow and its response to challenges is observed following administration of etomidate, ketamine, or a combination of low dose isoflurane/nitrous oxide and opioids. Thus, these anesthetics can be recommended for anesthesia in patients with already activated sympathetic outflow to maintain arterial blood pressure. PMID- 11413696 TI - [The significance of nitric oxide in nociception and spinal pain processing]. AB - Nitric oxide (NO) has been shown to be involved in the generation and processing of pain signals. Most experimental studies on animals and also the few observations in humans point to an involvement of the NO-system in inflammatory pain, whereas acute pain and chronic pain without inflammatory component seem to be independent of NO. It is yet unknown whether specific inhibition of the NO pathway is useful for treatment or prevention of inflammatory pain in humans. PMID- 11413697 TI - [How do we ethically justify non-resusitation? Arguments within the social system on an ongoing problem in modern medicine]. AB - The question whether to resuscitate or not belongs to the most controversial issues of medical ethics. This paper wants to give an insight into the German way of discussion on this topic. First it gives an overview of the most important codifications concerning resuscitation. Then it treats the question who is to be involved in do-not-resuscitate-decisions. The central point of the paper is the question of futile treatment and the question of the normative structures which are present in German discussion. PMID- 11413698 TI - [Droperidol and dimenhydrinate alone or in combination for prevention of postoperative nausea and vomiting]. AB - BACKGROUND: Droperidol and dimenhydrinate are inexpensive antiemetic drugs. Droperidol, especially, has been studied extensively in the past, but there are no studies that used the combination of both drugs for prevention of postoperative nausea and vomiting (PONV). Thus, the aim of this randomised controlled and double-blinded study was to evaluate the antiemetic efficacy and the side effects of such a combination therapy. METHODS: 240 inpatients undergoing ENT surgery under general anaesthesia were randomised to receive one of four antiemetic regimes: placebo, dimenhydrinate (1 mg x kg-1), droperidol (15 micrograms x kg-1), or the combination of both drugs (droperidol 15 micrograms x kg-1 + dimenhydrinate 1 mg x kg-1) was administered after induction of anaesthesia and repeated 6 hours after the first administration. For general anaesthesia a standardised technique including benzodiazepine premedication, propofol, desflurane in N2O/O2, vecuronium, and a continuous infusion of remifentanil was used. Postoperative analgesia and antiemetic rescue medication were standardised. Episodes of vomiting, retching, nausea, and the need for additional antiemetics were recorded for 24 hours. The main goal of the study was to increase the number of patients who were completely free from PONV (chi 2-test with Fisher-Yates' correction). Furthermore, the severity of PONV was analysed using a standardised scoring algorithm. RESULTS: Data of 227 patients could be analysed. The incidence of patients who suffered from PONV was 41.3% (95% confidence interval: 29-55%) in the placebo-group. Dimenhydrinate alone reduced PONV to 34.5% (95%-CI: 22-48%). This marginal effect and the effect of droperidol (PONV: 26.4% (95%-CI: 15-40%)) could not be proven statistically, since the power of the study was too small. The combination of both drugs decreased PONV to 19.6% (95%-CI: 10-32%) and also reduced the severity of the symptoms to a clinically acceptable level. CONCLUSION: Dimenhydrinate failed to reduce the incidence and severity of PONV. The efficiency of droperidol given alone was within the ranges previously known from metaanalytic data. The combination of both drugs showed a moderate synergistic effect. PMID- 11413699 TI - [The effects of high altitude on respiratory physiology]. PMID- 11413700 TI - [The effects of high altitude on the cardiovascular system]. PMID- 11413701 TI - [High altitude pulmonary edema (HAPE)]. PMID- 11413702 TI - [High altitude tolerance in patients with cardiac or pulmonary diseases]. PMID- 11413703 TI - [Successful cardiopulmonary resuscitation with a high-dosage bolus injection of rt-PA after fulminant pulmonary embolism]. AB - The fulminant pulmonary embolism is one of the most feared complications of hospitalized patients. A high percentage of those patients need resuscitation within a short period of time. In such cases, the thrombolysis is a quickly attained therapeutical alternative to pulmonary embolectomy. A 77-year-old man with a surgically managed femoral fracture who suffered a massive pulmonary embolism, had to be resuscitated on the 11th postoperative day. Because of the fast confirmation of the diagnosis with the ECG and transthoracical echocardiography, it was possible to initiate thrombolysis with a bolus of 100 mg rt-PA immediately after the beginning of cardiac arrest and resuscitation. After 20 min of cardiopulmonary resuscitation (CPR), the patient could be stabilized under high dosage of catecholamines. The patient survived after prolonged intensive care treatment without severe bleeding complications, neither did he present any neurological deficit. We conclude that in the case of massive pulmonary embolism with small chance of resuscitation, the high-dose bolus injection of rt-PA could enrich the therapeutical possibilities. PMID- 11413704 TI - [Chronic pain syndrome of the penis following cerebrovascular insult]. AB - Complex regional pain syndromes (formerly sympathetically maintained pain syndromes or reflex sympathetic dystrophy) encompass symptoms of pain, dysfunction and sympathetic disorder. They occur spontaneously or after peripheral or internal lesions (e.g. stroke or myocardial infarction) and predominantly affect the limbs, rarely the face or the trunk. This case report describes a 64-year old man who after a stroke suffered from heavy burning pain in the penis and perineum, which did not ameliorate after established conservative therapy. Sympathetic blockade, however, provided pain relief. The diagnosis of a complex regional pain syndrome, type I, was proposed according to the clinical symptoms in this patient, e.g. causalgia-like burning pain, allodynia, and the temporal association of an internal lesion to the onset of the pain. Other diagnoses such as neuropathic pain of unknown or diabetic etiology or a central post-stroke pain were considered. PMID- 11413705 TI - Application of an enzyme-immunoassay (EIA) for rapid screening of 5 alpha pregnane-3,20-dione (DHP) in blood plasma of the Asian elephant, Elephas maximus. AB - Populations of African (Loxodonta Africana) and Asian Elephants (Elephas maximus) in zoos and safari parks are at risk due to their low reproductive success. To extend the limited knowledge of their reproductive physiology, the development of easy and practical methods for the analysis of the relevant reproductive hormones is essential to support e.g. assisted reproduction. For the measurement of 5 alpha-pregnane-3,20-dione (DHP), the predominant ovarian gestagen in both species, an enzyme-immunoassay (EIA) based on commercial reagents was applied. Advantages of this EIA are the small volume of plasma needed for evaluation (5 microliters) and the possibility of direct processing without an extraction stage. The lower limit of detection was 0.16 ng/ml, the mean recovery was 101% and the mean coefficients of variation were 7.3% (intra-assay) and 9.9% (inter assay). In Asian elephants, DHP levels reached 15 ng/ml during the luteal phase and up to 21 ng/ml during pregnancy. Estrous cycle lengths based on the lowest DHP concentrations varied from 12 to 20 weeks (mean 15.4 +/- 2.3). In two Asian elephant cows a calf was still-borne. Thereafter, the animals reassumed ovarian activity after approximately 8 and 13 weeks, respectively. In one animal estradiol implants for hormonal contraception caused a down regulation of the ovarian function as demonstrated by an irregular pattern of DHP secretion over a period of 48 weeks. We propose the direct DHP-EIA as a suitable method for reproductive monitoring in elephants, as it can be easily established in laboratories. PMID- 11413706 TI - Histopathological studies on neoplasms in the genital tract of female donkeys in Egypt. AB - The genital tracts of 165 female donkeys were collected and examined with special concern to neoplasms. Ovarian Neoplasms were found in 12 cases (7.72%). A granulosa cell tumor was found in one case (0.61%). A cavernous haemangioma was found in 11 cases (6.67%). Uterine neoplasms were found in 14 cases (8.84%). Endometrial polyps and fibroleiomyoma were found in one case (0.61%) each. Leiomyoma was found in 2 cases (1.21%). Cavernous haemangioma was found in 10 cases (6.06%), and cervical polyps in 5 cases (3.03%). PMID- 11413707 TI - Mastitis in lactating camels (Camelus dromedarius) in Afar Region, north-eastern Ethiopia. AB - Quarter milk samples (n = 543) from 152 traditionally managed lactating camels (Camelus dromedarius) in Afar Region, north-eastern Ethiopia were examined to determine the prevalence of camel mastitis and identify its bacterial causes. Out of 152 camels examined, 19 (12.5%) were diagnosed as clinical mastitis cases based on clinical signs and bacteriological examinations. Of the 257 California Mastitis Test (CMT) positive quarter milk samples 162 (63.0%) yielded pathogenic bacteria. A positive correlation was observed between CMT positive results and presence of major pathogens in camel milk samples. The main mastitis pathogens isolated were Staphylococcus aureus, coagulase-negative staphylococci, Streptococcus agalactiae, S. dysgalactiae, and other species of streptococci, Pasteurella haemolytica and E. coli. Results of the present study suggest that mastitis in Afar camels is prevalent, Gram-positive cocci are the major isolates from camel milk samples and the CMT can be used as a screening test for the detection of mastitis in camels. PMID- 11413708 TI - Case report: entropion and periorbital dermatitis in an Assaf ram. AB - Entropion and periorbital dermatitis in an Assaf ram are reported. The severe clinical, pathological and histopathological findings are described. Parent animals of concern must be excluded from further breedings as the condition is inherited. The consequences of a such congenital condition in a ram and the recommendations are discussed. PMID- 11413709 TI - [Assessment of the Salmonella burden in slaughter pigs through the results of meat-juice-ELISA]. AB - During a study on the salmonella infection rate in slaughter pigs, bacteriological results were collected from faecal samples and intestinal lymphnodes, and serological results were acquired from meat-juice-ELISA. Serological findings and resulting serological parameters were analysed for their capacity to provide an indication of the salmonella burden in a given lot of slaughter pigs or in the farm of origin. The arithmetic mean antibody concentration of a lot of slaughter pigs or a piggery, or the number of animals reacting above 30% according to the Danish standard, or the number of animals reacting above 40% according to the Danish standard are all three equally suited to differentiate between a low and a high level of infection. The mean antibody concentration was found to be the best parameter for epidemiological studies that compare the serological status between herds or investigate the development of antibodies within a given piggery. PMID- 11413710 TI - [Increasing number of Salmonella paratyphi B isolates from slaughtered poultry sent in to the national Salmonella reference laboratory]. AB - In the last years the number of isolations of Salmonella enterica subspecies enterica serovar paratyphi B (S. paratyphi B) sent to the national salmonella reference laboratory of Germany has increased steadily. Most of the isolates originated from fowl or poultry products. The bacteriological, serological and biochemical properties of the isolates were investigated. Special emphasis was given to the utilization of d-tartrate which subgroups the serovar. All of them belonged to the d-tartrate positive variant, which is generally considered less virulent for humans and was formerly called S. java. The performance of various tests is compared and in addition the possibility of the spread within the production line is discussed. PMID- 11413711 TI - [Development of of non-immunising, paraspecific vaccine from attenuated pox viruses: a new type of vaccine]. AB - The various research periods leading to the development of paraspecific vaccines are described. Paraspecific vaccines are new, pyrogen-free, non-toxic preparations that contain non-immunising antigens and can be used to generate endogenic protective, non-antigen specific mechanisms in the sense of paramunization in humans and animals. They consist of highly attenuated and inactivated (0.05% beta-propiolactone) virus strains of various poxvirus genera. They activate the T helper cells and cellular elements of the paraspecific (innate) immune system and initiate the associated production and release of cytokines (cytokine cascade) with the goal of eliminating dysfunctions of the immune systems, rapidly enhancing the individual's non-pathogen- and non-antigen specific defences and exerting a regulatory effect on the interplay between the immune, hormone, nervous and vascular systems (signal-transduction mediators). They can be used systemically (intramuscularly) and locally (mucous membranes, skin). Immunization with paraspecific vaccines does not lead to postvaccinal complications and can be carried out as often as necessary, even for a number of years. They are compatible with conventional medicines and conventional specific vaccines. Closely linked protein complexes in the envelopes of the virus particles are responsible for their efficacy some of those envelope protein complexes possess the properties of weak super antigens. Paraspecific vaccines have proved effective in combating viral infections, in particular herpes and hepatitis B and C infections, and chronic inflammatory diseases, and also as adjuvant therapy for tumours, for curing stress-related disturbances and dysfunctions of the immune system. PMID- 11413712 TI - The effect of different quantities and compositions of pelleted diets on immune response of mares during the production of anti-tetanus sera. AB - Research was carried out into the effect that different quantities and compositions of concentrated portions of meal had on certain haematological properties and on the immune response of mares in the course of hyper-immune antitetanus sera production. The experiment involved 24 Nonius and Lipizzaner cross-bred mares divided into two groups of 12 animals each, a control group and a trial group. The experiment lasted 12 months, with haematological and immunological tests being carried out every 30 days. During the course of the experiment each mare was subjected to 11 immunisation cycles, and in that period no mare fell ill. Leukocyte and haematocrit counts revealed no differences between the control and the test groups throughout the entire duration of the experiment. Following the second immunisation cycle the leukocyte count increased in both groups of mares, remaining significantly higher than the normal physiological value in horses throughout the experiment. The concentrated portion of meal, differing both in quality and quantity, had no influence on leucocyte count and haematocrit level in the mares throughout the experiment. However, the immunodiffusion titre in the test group did show a slightly higher value of ID titre than in the control group, indicating that the quantitatively and qualitatively differing portions of meal had a more positive effect on immune response and anti-tetanus anti-body titre in the test group than it did in the control group. PMID- 11413713 TI - [Liver fluke infection and sarcoptic mange in red foxes in Berlin]. AB - 396 red foxes originating from the city of Berlin were examined for opisthorchiid liver flukes and clinical sarcoptic mange between January 1997 and March 1998. Out of 232 (= 58.6%) foxes positive for opisthorchiid flukes 221 animals harboured Metorchis bilis and 70 were infected with Opisthorchis felineus. Pseudamphistomum truncatum was found only in 8 foxes. M. bilis occurred as mono infection in 154 animals. M. bilis in combination with O. felineus was found in 61 cases. Pure Opisthorchis infection as well as other fluke combinations were found in a small number of animals only. 85 (= 21.5%) foxes showed clinical sarcoptic mange. Liver fluke positive foxes showed a higher mange prevalence than uninfected animals. However, significant associations between flukes and manage were only found when comparing uninfected foxes with those having the highest worm burden. The association of liver flukes and mange could be established for adult female foxes by a significant Odds Ratio of 4.3. PMID- 11413714 TI - [The diagnostic value of venous blood gas parameters and pH value in newborn foals with pulmonary diseases]. AB - Analysis of blood gases in equine neonatology is regarded as a diagnostic tool to study the neonatal adaptation period. Aim of this study therefore was to compare the diagnostic value of venous blood gas parameters to arterial parameters in newborn foals with pulmonary disorders. Venous as well as arterial blood samples were taken from 24 foals (1 to 6 days old) and the partial pressure of oxygen (pO2), partial pressure of carbon dioxide (pCO2), pH, and oxygen parturition (S O2) of these samples were investigated. In addition, the alveolar (A) to arterial (a) gradients (A-aDO2) were calculated. Due to changes in blood gas parameters during the first week postnatal the age was taken into consideration by using covariance analysis. All arterial parameters except paCO2 showed a significant difference among healthy foals (n = 15) and foals with respiratory disorders (n = 11) with A-aDO2 and paO2 being the most reliable arterial parameters. In venous blood there was a significant difference between healthy and sick foals only in S O2 and pH. PMID- 11413715 TI - [Sonography of the teat of cattle, sheep and goats in comparison with other methods of diagnostic imaging--a review]. AB - Beside the clinical examination of udder/teat imaging methods e.g. sonography, radiography and endoscopy allow diagnosis. This publication will give a summary of currently used and potentially usable methods of diagnostic imaging applied to the teat. Advantages and disadvantages of the several methods were compared--as quoted in literature and completed by own experiences. PMID- 11413716 TI - [Detection of levamisole and ivermectin in organ samples from a dead collie]. AB - A collie, known for its breed-dependent adverse reaction to ivermectin, was without any clinical signs. The dog was prophylactically treated with 3 mg/kg KG (s.c.) of levamisole. Within 15 minutes, the dog showed convulsions, vomitus, and dyspnea, and perished 2.5 hours after injection of the drugs. The pathological findings were not informative as to the cause of death, and with regard to the adverse reactions, additional application of ivermectin was not excluded. Therefore, organ samples were submitted for toxicological analysis of both levamisole and ivermectin. For detection of levamisole and ivermectin, modified GC/MS and HPLC procedures were developed. Concentrations up to 535 micrograms levamisole and up to 26 ng ivermectin were found per g tissue. Both analytical methods are sensitive enough to detect these drugs after application of low doses. This study elucidates that combination of low-dosed ivermectin and levamisole is no recommendable means against adverse effects of ivermectin, with respect to collies. Moreover, the synergistic effects of ivermectin and levamisole suggests the same drug incompatibility in other dog breeds and animal species. PMID- 11413717 TI - Lead poisoning in a dog--a case report. AB - Lead poisoning in a bitch is described. Clinical signs, hematological and biochemical changes, X-ray, as well as presence of lead in stomach and blood confirmed the diagnosis. PMID- 11413718 TI - [Recent plant poisoning in ruminants of northern and eastern Germany. Communication from the practice for the practice]. AB - It is reported of botanical intoxications in 1379 ruminants (dairy cattle, beef cattle and ewes) in north-eastern Germany. The intoxications of these animals were caused by consumption of the following plants: meadow saffron (Colchicum autumnale L.), cowbane (Cicuta virosa L.), sweet clover (Melilotus alba MED.), sheep's sorrel (Rumex acetosella L.), bracken (Pteridium aquilinum KUHN) and St. John's wort (Hypericum perforatum L.). In all the five described cases animals died as a result of plant intoxications. The authors come to the conclusion that periodical inspections of the meadows are necessary, checking the plants in cooperation with veterinary surgeons. PMID- 11413719 TI - Occupational stress and psychiatric illness in the military: investigation of the relationship between occupational stress and mental illness among military mental health patients. AB - OBJECTIVE: This was a pilot study aimed at gathering preliminary data on the relationship between occupational stress and mental illness among military personnel. The primary goal of this study was to determine to what extent military mental health patients report suffering from significant occupational stress. METHODS: Eighty-five active duty military mental health outpatients at the Wilford Hall U.S. Air Force Medical Center mental health clinic answered a 65 item survey that included items on the perception of occupational stress and reported life changes. Participation in this cross-sectional study was anonymous, voluntary, and random. The 85 participants represent 83% of those surveyed and approximately 10% of the clinic's total population of military mental health outpatients. The survey incorporated the 43-item Schedule of Recent Experiences (SRE). By adding the weighted values assigned to the 43 items, each respondent was given an SRE score, which is a measure of overall stress that has been shown to be predictive of future illnesses. RESULTS: A majority (60%) reported suffering from significant work stress. A majority (52%) reported that work stress was causing them significant emotional distress. Almost half (42.5%) reported that work stress was a significant contributor to the onset of their mental illness. The average SRE score for all respondents was 266, reflecting increased risk for future illnesses. Generic work stressors were endorsed more frequently than military-specific stressors. CONCLUSIONS: The results reveal that this population of military mental health clinic outpatients perceived that work stress had a strong negative effect on their emotional health. These results raise the possibility that work stress could be a significant occupational health hazard in the U.S. military, a possibility that warrants further investigation. By gathering additional data on the relationship between work stress and emotional health in the military, interventions can be planned to mitigate the effect of stress caused by the military work environment on the mental health of military personnel. PMID- 11413720 TI - Analysis of environmental issues for nursing aboard the USNS Mercy (T-AH 19). AB - The USNS Mercy (T-AH 19) is a unique environment for its staff and patients. Several environmental influences distinctively affect health, medical, and nursing care. Six main areas of environmental concern for operational nursing were examined. The strengths, weaknesses, opportunities, and threats to the internal environment of a hospital ship were explored. Findings indicate that the USNS Mercy meets or exceeds many environmental standards. However, there is significant room for improvement and nursing involvement in issues related to the management of infectious diseases, patient transport, care of patients with nuclear, biological, or chemical agents, and management of wastes aboard the ship. Nursing implications for research and practice are proposed. PMID- 11413721 TI - Health assessment of U.S. military personnel deployed to Bosnia-Herzegovina for operation joint endeavor. AB - In anticipation of U.S. forces deploying to Bosnia-Herzegovina, plans were established to conduct medical surveillance of all military service members. This surveillance would provide the Department of Defense with an overview of the hospitalization and outpatient morbidity experience of U.S. forces. Standardized collection of medical data from all U.S. camps using 14 diagnostic categories based on International Classification of Diseases, 9th Revision, codes began in March 1996. Special assessments for hantavirus and tick-borne encephalitis (TBE) infection risk were also conducted. The average disease and nonbattle injury rate for U.S. forces was 7.1 per 100 soldiers per week. Injuries accounted for 28% of medical visits, whereas undefined/other visits accounted for 33%. The majority of remaining visits were for respiratory (14%), dermatologic (10%), and gastrointestinal (6%) complaints. There was one confirmed and one suspected case of hemorrhagic fever with renal syndrome; only 0.1% of individuals (2 of 1,913) tested seroconverted to hantavirus during deployment. No cases of TBE were reported, and the overall low seroconversion rate (0.42%, 4 seroconversions among 959 unimmunized personnel) reflected a very low risk of infection with TBE related viruses. Operation Joint Endeavor and follow-on Operations Joint Guard and Joint Forge have been extremely healthy deployments. PMID- 11413722 TI - Quality of deployment surveillance data in southwest Asia. AB - The Global Expeditionary Medical System (GEMS), formerly known as Desert Care II, provides clinical data on every medical encounter that occurs at U.S. Air Force medical treatment facilities in theater. After 22 months of surveillance from March 1997 to January 1999, 59,026 records were generated from 27,305 active duty members. A random sample of 273 individuals were reviewed for four key fields: chief complaint, diagnosis, International Classification of Diseases, 9th Revision, code, and disease and nonbattle injury category. The vast majority (> 99%) of records were consistently classified and reported. An unrelated subset of all cases categorized as "Medical/Other" seen at Prince Sultan Air Base (5,640 records) also were analyzed. Reassignment was made in 19% of cases into other categories, suggesting that further refinement of disease and nonbattle injury reporting is needed for effective deployment medical surveillance. Overall, this electronic data-gathering system has high internal validity, consistency, and reliability for service members in the deployed U.S. Air Force setting. PMID- 11413723 TI - Etiological factors of mental disorders among members of the Polish military forces. AB - OBJECTIVE: This project attempts to establish possible links between psychosocial factors and mental disorders among Polish professional soldiers. MATERIALS AND METHODS: The examined group consisted of 1,785 professional soldiers hospitalized in the psychiatric department of the 107th Military Hospital in Walcz in the years 1982 to 1984, 1986 to 1989, and 1993 to 1996. The study was based on analysis of accessible medical files. Special attention was paid to diagnosis, age, year of service, garrison size, position, and motivation to remain in military service. The International Classification of Diseases, Tenth Revision, diagnostic scheme was used. RESULTS: Neurotic disorders were the most often diagnosed psychic disturbances in all groups of professional soldiers. More than half of the soldiers hospitalized from 1986 to 1989 became unfit for further military duty because of their mental disturbances. This study confirmed statistical links between dismissals of professional soldiers and motivation, age, or job seniority. Also, we found a link between military positions and increased psychiatric morbidity among soldiers. Personality disturbances, maladaptive syndromes, and substance-related disorders dominated at command positions in all years investigated. CONCLUSION: The results showed the necessity of further elaborations of preventive and new treatment rules, leading to a diminution of mental disturbances among professional soldiers in Poland. PMID- 11413724 TI - Soldier and family wellness across the life course: a developmental model of successful aging, spirituality, and health promotion. Part I. AB - The primary purposes of this article are to (1) highlight current challenges facing health promotion advocates within the military and civilian culture; (2) present the strengths and weaknesses of the current Army approach to health promotion and preventive medicine; and (3) present several unifying themes that contribute to enhanced progress within the field of health promotion. A conceptual model that links common goals across the fields of successful aging, health promotion, spirituality and health, and life course is advocated to maximize efficacious interventions and to transform the current Army approach to health promotion. A companion article will describe an integrative model of health promotion and wellness that responds to the challenges and incorporates the unifying themes described in this article. PMID- 11413725 TI - Military, civilian, and rural application of the damage control philosophy. AB - Damage control surgery is a useful salvage strategy for the most critically injured patients. Conceptually, this approach to individual patients can be extrapolated to situations such as military field surgery, civilian mass casualty events, and long-range transfers from rural areas. The logistic realities of Army forward surgery teams are addressed with regard to typical damage control maneuvers and evacuation. Specific areas requiring improvement through directed research are identified. Initial civilian mass casualty strategies are discussed, and a plan to prevent transfer delays of rural trauma patients is presented. By transferring the lessons learned from individual damage control patients to military, civilian mass casualty, and rural casualty events, resource utilization is optimized. The concept of minimal acceptable care rather than optimal trauma care can be applied to these three seemingly different situations. PMID- 11413726 TI - Assessing myocardial viability using the coronary flow response to intravenous dobutamine infusion in recent myocardial infarction. AB - Coronary flow response to low-dose (5 and 10 micrograms/kg/min) dobutamine infusion was used to assess myocardial viability at the time of cardiac catheterization in 13 patients (age, 60 +/- 11 years) with recent myocardial infarction. Echocardiographic improvement in regional wall motion performed 4 to 6 weeks after discharge was used as the marker for viability. Viable patients demonstrated a 2-fold increase in flow from baseline (p < 0.001) during intravenous infusion. In contrast, patients without viability demonstrated no increase in flow. The coronary flow response to dobutamine measured at the time of catheterization shows promise in identifying viable myocardium in postinfarction patients. PMID- 11413727 TI - Five-year experience with implantation and follow-up of transvenous implantable cardioverter defibrillators: placing postimplant defibrillation threshold testing in perspective. AB - BACKGROUND: The rapid technological advancement in transvenous implantable cardioverter defibrillators (ICDs) has resulted in heterogeneous and often controversial approaches to follow-up procedures. The efficacy of postimplantation defibrillation threshold (DFT) testing with new-generation biphasic ICDs is unknown. OBJECTIVE: In this retrospective study, predischarge and postdischarge DFT protocols were compared to evaluate their safety and effect on adverse clinical events. METHODS: The study population consisted of 89 patients with 92 ICDs and 103 endovascular lead systems. Forty-four patients had DFT tests during implant and the predischarge period. Thirty patients had DFT tests during implant and the postdischarge period. Sixteen patients had only implant DFT data available. The follow-up period ranged from 3 days to 5.6 years. RESULTS: Ninety-nine percent of patients had successful implants. Postimplant DFT tests detected potential problems in only 1% of asymptomatic patients. Thirty-six percent of patients with normal predischarge DFT tests had adverse clinical events compared with 18% in the postdischarge group. Patients with postimplant DFTs > 25 joules (J) and safety margins > or = 10 J had a lower incidence of adverse clinical events (p = 0.03) compared with those with safety margins < 10 J. An 11% malfunction rate was observed in ICD leads during the follow-up period. CONCLUSIONS: DFT testing after implant is safe; however, routine postimplant DFT testing has limited value in assessing abnormalities in patients with the current generation of biphasic transvenous ICD devices. A 10-J safety margin was associated with a lower incidence of adverse clinical events in patients with DFTs > 25 J. Endovascular lead failure remains a significant problem with ICD systems requiring vigilant follow-up. PMID- 11413728 TI - "Tip of the spear" physical therapy during a 6-month deployment to the Persian Gulf: a preliminary report. AB - When a U.S. Navy aircraft carrier battle group deploys overseas, the ship's medical department is responsible for more than 10,000 personnel and their numerous musculoskeletal injuries. This paper reviews the effectiveness of having a U.S. Navy physical therapist and physical therapy technician onboard the USS Carl Vinson during its most recent deployment to the Persian Gulf. Physical therapy had 3,373 patient visits during the ship's 1998-1999 Western Pacific deployment. Having physical therapy personnel onboard resulted in fewer patient visits to sick call for musculoskeletal problems and fewer evacuations compared with other similar carrier deployments. Providing physical therapy at the "tip of the spear" is an effective, beneficial, and cost-saving landmark improvement in providing quality medical care to the fleet. The lessons learned from this experience will assist in clarifying the role of physical therapy in future military support operations and sustained deployments. PMID- 11413729 TI - Health-related quality of life in Persian Gulf War Veterans. AB - OBJECTIVE: The objective of this investigation is to describe the health-related quality of life of Persian Gulf War (GW) veterans and to examine the effects of current chronic medical conditions and psychiatric status on physical functioning. METHODS: To measure health-related quality of life, the Medical Outcomes Short Form Survey (SF36) was administered approximately 4 years after the GW to a stratified, random sample of New England-area GW-deployed veterans and a group of military personnel deployed to Germany during the GW. The SF36 scores for the GW-deployed study population (N = 141) were compared with those for the Germany-deployed group (N = 46) and with published U.S. population norms. Multiple linear regression analyses were performed to identify risk factors associated with lower physical health functioning in the GW-deployed study group. RESULTS: Functional health status was significantly lower in the GW-deployed group compared with the Germany-deployed group for each of the SF36 subscales and the two summary scores (Physical Component Summary [PCS] and Mental Component Summary). Compared with the general U.S. population, the GW-deployed group median was between the 25th and 50th percentile for the Physical Functioning subscale and the PCS score. Within the GW-deployed group, lower education, psychological symptomatology, and a higher number of chronic self-reported medical conditions were significant predictors of the PCS score. CONCLUSION: GW-deployed veterans report lower functional health status compared with a group of Germany-deployed veterans and published general U.S. population norms. Within the group of GW deployed veterans, several current medical and psychological conditions predictive of lower physical functioning levels were identified. PMID- 11413730 TI - Correlates of physical fitness improvement among a Navy remedial exercise sample. AB - This study investigated influences on physical fitness improvement among Navy remedial exercise program participants. Subjects were 27 Navy personnel who had failed to pass the body composition, aerobic fitness, or curl-up portion of the semiannual physical readiness test. The remedial exercise program consisted of three 1-hour exercise sessions per week for 16 weeks, with primary emphasis on running and walking. To determine influences on physical fitness improvement, physical readiness test and self-report survey data were collected and analyzed through bivariate Pearson correlation. No correlations were found between any of the study variables and weight change. Initial body fat was positively correlated (r = 0.470, p < 0.05), whereas initial body weight (r = -0.490, p < 0.05) and perceptions of exercise leadership (r = -0.690, p < 0.05) were negatively correlated with reductions in body fat. Positive correlates of improvements in 1.5-mile run time were initial run failure (r = 0.637, p < 0.01), perceived exercise benefits (r = 0.574, p < 0.01), and perceptions of social support (r = 0.543, p < 0.05). Exercise mode (walking group) was negatively correlated (r = 0.445, p < 0.05) with improvements in run time. In conclusion, this study demonstrated that various personal, program, and environmental factors influence improvements in physical fitness status. PMID- 11413731 TI - A survey of Navy physicians' attitudes toward the use of selective serotonin reuptake inhibitors in active duty military personnel. AB - A series of recent pharmacological discoveries have produced remarkable advances in psychotherapeutic medications. With the advent of newer antidepressants, there is a high degree of efficacy and a low risk profile. Potential benefits of these medications far outweigh the possible side effects, especially in contrast to older drugs. Navy Medicine now has the ability to treat active duty personnel during a deployment with safe antidepressant medications. The attitudes toward the use of selective serotonin reuptake inhibitors (SSRIs) on active duty service members has been surveyed. Profiles of prescribing patterns and attitudes toward the use of these medications and safety within the context of the operational environment were also surveyed. Group comparisons across various medical specialties and command organizations were made. The survey results suggest a very favorable attitude toward prescribing SSRIs in the active duty population. PMID- 11413732 TI - Depression and torture. AB - Torture represents an exceptionally traumatic experience in which horror, helplessness, and hopelessness are extreme. Therefore, it can be expected that depression, along with other trauma-related disorders is present in torture victims at higher rates than in other psychotraumatized individuals. To demonstrate this, we examined two groups of refugees, all suffering the post traumatic stress disorder. The first group (N = 50) had combat experience but were imprisoned and tortured as well. Members of the second group (N = 29) had combat experience. A third group (N = 30) consisted of local people with no traumatic experience. Using the Hamilton scale, the Beck Depression Inventory and structured dedicated interviews, we tried to determine whether those groups differed in level of depression based on their different levels of traumatic experience. The results of our study indicate that torture victims showed a significantly higher level of depression that is clinically relevant. PMID- 11413733 TI - Accidents and injuries among U.S. Navy crewmembers during extended submarine patrols, 1997 to 1999. AB - Accidents and injuries, the most common cause of morbidity in military populations, result in a significant number of work days lost each year and account for 75% of all active duty deaths. Rates of accidents and injuries during U.S. Navy submarine deployments have not been evaluated previously. A database designed to monitor the health of submarine crew-members was used to examine the rates and causes of accidents among deployed crewmembers aboard 196 submarine patrols between 1997 and mid 1999. The most common category of injuries was open wounds, followed by sprains and strains, contusions, superficial injuries, burns, and others. Rates of accidents and injuries decreased with increasing age and duration of military service. Among submariners working in supply departments, the rates were more than two times those of crewmembers working in other departments. Based on these data, among a submarine crew of 100 men at sea for 100 days, approximately four to five accidents or injuries might be expected and would result in an average of about 2 days of light or no duty per injury. Rates of accidents and injuries were very low; however, focused safety training could reduce rates among younger and less experienced crewmembers as well as among those working in particular areas of the submarine. PMID- 11413734 TI - Predicting dental treatment workload of U.S. military personnel. AB - As the demographic composition of the U.S. armed forces shifts, so does dental treatment workload (DTW). To date, the effects of such shifts on DTW have been speculative. This study seeks to build regression models to predict the DTW of recruit and active duty military personnel as service demographic factors shift. The data come from a 1994-1995 random sample of 13,050 active duty and 2,711 recruit personnel. Dental treatment needs were charted for all participants. Patient demographic variables--age, sex, race, education, branch of service, and annual dental utilization (for all); home region and marital status (recruits only); and rank (active duty only)--were noted. Treatment needs were converted to composite time values. Respondent demographic factors were regressed on composite time values to determine which factors influence DTW using backward, stepwise, linear regression. Results show that DTW varies across demographic categories. The magnitude and direction of change in DTW as service demographic variables shift can be predicted with linear regression models. PMID- 11413735 TI - Infectious disease rates in the U.S. Navy, 1980 to 1995. AB - The effect of increasing numbers of women in the U.S. Navy, particularly those aboard Navy ships, on infectious disease risk is unknown. This study examines gender and other demographic differences among all U.S. Navy enlisted personnel in first hospitalizations for infectious diseases from 1980 through 1989 and identifies trends in incidence rates during the extended period from 1980 to 1995. All data were obtained from official personnel and medical records. First hospitalization rates were computed using the Epidemiological Interactive System. Varicella and other viruses and chlamydiae accounted for more than 20,000 hospitalizations among Navy enlisted personnel in the 1980s. In 7 of the 12 categories of common infectious diseases, women's rates were higher than those for men, particularly for viral meningitis, herpes simplex, syphilis, gonococcal disease, and candidiasis. An excess of certain common infectious diseases among women and nonwhite ethnic groups emphasizes the need for continuing education and surveillance in these populations. PMID- 11413736 TI - Contraceptive use and attitudes toward family planning in Navy enlisted women and men. AB - Little is known about the contraceptive behavior and beliefs of Navy personnel. Nevertheless, the Navy, in its role as primary medical provider for its personnel, needs to know whether sailors have access to effective birth control and are sufficiently informed about contraception to make wise choices. As part of the Women Aboard Navy Ships Comprehensive Health and Readiness Project conducted at the Naval Health Research Center in San Diego, California, contraceptive use and attitudes toward family planning were assessed through a survey administered to 714 enlisted women and 665 enlisted men on 15 ships. Contraceptive use was not related to gender, age, marital status, pay grade, race, or education. More favorable family planning attitudes were related to contraceptive use. Women and men differed in their attitudes toward family planning, with women's responses more positive than men's. The results highlight the need for research focusing on the effect of attitudes on contraceptive behavior. PMID- 11413737 TI - An accidental death caused by an unexploded 40-mm grenade. AB - Unexploded grenades are potentially dangerous materials. Numerous grenades have been found to be unexploded after wars have ended. We present an interesting case of an unexploded grenade that was found accidentally in a soldier's skull. On physical examination, the entrance wound of an unusual penetrating object was noted. This unusual physical finding led us to undertake radiographic examination. A 40-mm unexploded grenade was noticed on the radiograph of the soldier's skull. PMID- 11413738 TI - Relative importance of calcium-activated potassium channels in nipradilol-induced aortic relaxation in rats. AB - Nipradilol (CAS 81486-22-8), a vasodilatory beta-blocker, has been shown to dilate smaller vessels than nitroglycerin does, and the vasodilative effects of nipradilol have been reported to be less mediated by cyclic GMP (guanosine monophosphate) than those of nitroglycerin. To test the hypothesis that cyclic GMP-independent potassium channels have a larger role in nipradilol-induced aortic relaxation than cyclic GMP-dependent mechanisms, the effects of a potassium channel blocker, tetraethylammonium (TEA, CAS 56-34-8), and of a guanylate cyclase inhibitor, methylene blue (MB, CAS 61-73-4), on nipradilol induced aortic relaxation were investigated and compared with those on nitroglycerin-induced aortic relaxation in isolated rat aortic rings. Relaxation response was expressed as percent relaxation, which is a percentage of the tension developed by 10(-7) mol/l norepinephrine. Nitroglycerin and nipradilol similarly relaxed the aortic ring in a concentration-dependent manner (10(-9)-10( 4) mol/l). In contrast, desnitronipradilol, a nipradilol analogue which has no nitroxy group, induced almost no aortic relaxation. TEA at 10(-3) mol/l, which is selective for calcium-activated potassium channels, inhibited the aortic relaxation induced by nipradilol (10(-5) mol/l) to a significantly greater extent than that induced by nitroglycerin (10(-5) mol/l) (% relaxation: 30.0 +/- 6.8 vs. 51.1 +/- 6.1%, p < 0.05). MB (10(-5) mol/l) suppressed the relaxation by nitroglycerin slightly but not significantly more than that by nipradilol. (% relaxation: 54.7 +/- 9.9 vs. 64.6 +/- 5.7%). The combination of TEA and MB almost completely eliminated the relaxation induced by nipradilol as well as by nitroglycerin. Thus, cyclic GMP-independent calcium activated potassium channels may be more involved in the aortic relaxation by nipradilol than that by nitroglycerin in rats. PMID- 11413739 TI - Comparative efficacy and safety of azelastine and levocabastine nasal sprays in patients with seasonal allergic rhinitis. AB - The aim of the present investigation was to compare the efficacy and tolerability of azelastine (CAS 58581-89-8) (1.12 mg/day) and levocabastine (CAS 79547-78-7) (0.4 mg/day) nasal spray administered twice daily to patients with seasonal allergic rhinitis. A total of 180 patients participated in a 4-week, double blind, parallel group (n = 90 each) study. Symptom severity of nasal, ocular and other symptoms were recorded, out of which a total symptom score (TSS) was calculated. Physicians assessed symptoms at baseline and at days 7, 14, and 28, patients and physicians evaluated the efficacy and tolerability. After 4 weeks of treatment with azelastine the mean overall TSS was reduced from a baseline score of 18.7 to 4.2, after levocabastine from 17.8 to 5.9. Patients morning scores for treatment days 1 to 28 gave a mean total score of 212.4 for the azelastine group and 230.6 for the levocabastine group; the equivalent evening scores yielded a mean total score of 115.5 and 175.6 respectively. Global efficacy was judged by physicians as either 'very good' or 'good' for 90% of azelastine patients and for 74% of the levocabastine group; 92% of azelastine patients and 76% of levocabastine patients judged treatment to be either 'very good' or 'good'. No serious adverse events were reported, all adverse events were related to nasal symptoms. Both azelastine and levocabastine administered twice daily as a nasal spray provide effective and well tolerated symptomatic treatment of seasonal allergic rhinitis. Azelastine, however, was statistically superior in efficacy as well as in safety (PWei-Lachin < 0.0001, combined results). PMID- 11413740 TI - Pharmacokinetics of NS-49, a phenethylamine class alpha 1A-adrenoceptor agonist. 4th communication: tissue distribution, placental transfer and milk secretion of radioactivity in rats after a single oral administration of 14C-NS-49, and effects of repeated administration on its pharmacokinetics. AB - The tissue distribution, placental transfer and milk secretion of 14C-NS-49 ((R) (-)-3'-(2-amino-1-hydroxyethyl)-4'-fluoro-methanesulfonanilide hydrochloride, CAS 137431-04-0), a phenethylamine class alpha 1A-adrenoceptor agonist, have been studied after a single oral administration (1 mg/kg) of a suspension formulation to rats. Radioactivity concentrations in tissues were generally highest 1 or 4 h, and for most tissues, exceeded those in the corresponding plasma. Concentrations were generally similar in male and female rats and persisted for at least 24 h. Radioactivity concentrations in most tissues declined in parallel with those in plasma. Placental transfer of radioactivity was low accounting for < 0.1% of the maternal dose. In milk, concentrations were of a similar order to those in the plasma but reached a peak later: the data implied that 14C-NS-49 readily diffused from the plasma into the milk. The absorption, distribution and excretion of 14C NS-49 have been studied after the repeated administration (1 mg/kg) of a suspension formulation to rats for up to 21 days. At 21 days, radioactivity concentrations in plasma reached a peak 1 h and declined with a terminal half life of 67 h. Steady state concentrations were reached during 14 days. Peak concentrations in tissues occurred 1 h and, in most tissues exceeded the plasma value. Radioactivity concentrations in tissues appeared to reach steady state during the 21-day dosing period. Tissue and blood cell concentrations declined more slowly than those in the plasma. Radioactivity excretion was relatively constant during the repeated administration and similar in urine (mean 45.8% total dose) and feces (mean 48.2% total dose). At 7 days after the last of 21 daily oral doses, only 0.2% of the total dose remained in the body, indicating that there is no marked accumulation of radioactivity in the tissues. The results obtained in these studies indicated that rats receiving NS-49 at 24 h intervals during chronic and reproductive toxicity studies would be continually exposed to the parent compound and/or its metabolites. PMID- 11413741 TI - Tissue distribution of NS-49, a phenethylamine class alpha 1A-adrenoceptor agonist, in pigmented rats. AB - After a single oral administration of 1 mg/kg of 14C-NS-49 ((R)-(-)-3'-(2-amino-1 hydroxyethyl)-4'-fluoromethanesulfonanilide hydrochloride, CAS 137431-04-0), the radioactivity distribution in tissues of male pigmented rats was studied and compared with that in male albino rats. One eye of each pigmented rat was divided into melanin-containing structures (uvea, pigmented epithelium and sclera) and others without melanin (cornea and lens), and the radioactivity concentration in each ocular tissue was measured. In all the pigmented rat body tissues tested, maximum radioactivity concentrations (Cmax) were reached within 4 h after administration. At 1 h, the kidney showed the highest concentration (11 times the plasma concentration), followed by the urethra, liver, urinary bladder and lung. Concentrations in the other tissues were similar to or less than the plasma concentration. The radioactivity concentrations in most tissues decreased rapidly, being less than 10% of the Cmax 24 h after administration. These characteristics of the 14C-NS-49 tissue distribution in the pigmented and albino rats did not differ. In the eye, however, the radioactivity concentration decreased more slowly in the pigmented than in the albino rats. Most of the radioactivity in the eyes of the pigmented rats was present in melanin-containing structures indicating that NS-49 binds to ocular melanin. The radioactivity concentration in the melanin-containing structures reached a maximum 4 h after administration, then decreased as did that for the whole eye with a t1/2, beta of 66.8 h. PMID- 11413742 TI - Transdermal delivery of nalbuphine and nalbuphine pivalate from hydrogels by passive diffusion and iontophoresis. AB - The objective of this study was to evaluate the in vitro transdermal permeation of nalbuphine hydrochloride (CAS 23277-43-2) (NA) and nalbuphine pivalate (NAP), a novel prodrug of NA, from different hydrogel formulations under passive diffusion as well as iontophoresis. Various concentrations of polymers, including polyvinylpyrrolidone (PVP) and hydroxypropyl cellulose (HPC) were used in the hydrogel formulations. The passive permeation rate of NA was affected by the polymer concentrations, which can be attributed to different viscosities of the hydrated formulations; whereas the passive permeation rate of NAP was not influenced by the various polymer concentrations. Iontophoresis significantly increased the permeation rates of NA and NAP from various hydrogel formulations through skin; the enhancement ratios were higher for NA in all the formulations studied. The iontophoretic permeation rates of NA were slightly decreased by the incorporation of polymers; however, the transdermal flux and membrane potential were independent of polymer concentrations for both NA and NAP, demonstrating that the polymer concentrations in the hydrogel formulations did not have significant effects on the iontophoretic permeation of NA and NAP. PMID- 11413743 TI - Immunological safety evaluation of a horse collagen haemostatic pad. AB - The safety of a haemostatic and wound dressing collagen pad (Antema) obtained from horse Achilles tendon treated with pepsin, has been studied from the immunological point of view. A sensitizing activity test and maximization test in guinea pig has been performed and its capacity of inducing antibodies has been tested in rabbit and man. In guinea pig the collagen pad tested did not show any symptom of allergic reaction, in rabbit it induced a weak immunological response, without any clinical symptoms, detectable with the ELISA method only at very high serum concentration levels (1:20). In a clinical trial, 20 patients submitted to different types of surgery in which the product had been used as haemostatic pad and left in the surgery area until absorption, did not develop any immunological clinical reaction. One year after the operation no traces of anti-collagen antibodies have been found with the ELISA method in the sera of those patients. PMID- 11413744 TI - Synthesis of some benzimidazole derivatives and their antibacterial and antifungal activities. AB - A number of new benzimidazole derivatives were synthesized by the reaction of benzimidazole with appropriate alkyl halides. The compounds synthesized were intensified by 1H-NMR, Fourier Transformation Infrared (FT-IR) and micro analysis. All new and related compounds studied in this work were screened for their in vitro antimicrobial activities against the standard strains: Enterococcus faecalis (ATCC 29212), Staphylococcus aureus (ATCC 29213), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853) and the yeasts Candida albicans and Candida tropicalis. Eleven of the compounds were found effective to inhibit the growth of Gram-positive bacteria (E. faecalis and S. aureus) at MIC values between 12.5-400 micrograms/ml. None of the compounds exhibited antimicrobial activity against Gram-negative bacteria (E. coli and P. aeruginosa) at the concentrations studied (6.25-800 micrograms/ml). All compounds (except compound 3) were significantly effective against C. tropicalis with MIC values of 6.25-400 micrograms/ml. Eight of the tested compounds showed an antifungal activity against C. albicans with a range of the MICs between 50 and 400 micrograms/ml. PMID- 11413745 TI - Four-week oral toxicity studies of the new quinolone antibacterial agent cetefloxacin tosylate in rats and marmoset monkeys. AB - Four-week oral toxicity studies with cetefloxacin tosylate ((-)-7[3-(R)-amino-2 (S)-methyl-1-azetidinyl]-1-(2,4- difluorophenyl)-1,4-dihydro-6-fluoro-4-oxo-3 quinolinecarboxylic acid tosylate, CAS 141725-88-4 (base), E-4868.Ts) a new quinolone antibacterial agent, were performed in Sprague-Dawley rats and marmoset monkeys at doses of 100, 450, 2000 mg/kg/d and 25, 50, 125, 300 mg/kg/d, respectively. In rats, due to its toxicity the high dose was decreased to 1000 mg/kg/d after 3 days of treatment. Mortality was recorded among high dose rats receiving 2000 or 1000 mg/kg/d. Rats receiving dosages of 450 or 2000/1000 mg/kg/d showed less activated mandibular lymph nodes, cortical lymphocyte depletion of mandibular and/or mesenteric lymph nodes, atrophy of the white pulp of the spleen, cortical atrophy of thymus and thymic apoptosis. Enlarged caeca, increased water consumption and variations in plasma electrolyte levels were observed in animals receiving these dosages and in male rats receiving 100 mg/kg/d. Low neutrophil counts were observed in rats receiving dosages of 100 or 450 mg/kg/d, and increased alkaline phosphatase and alanine transaminase plasma levels and slightly decreased plasma protein levels in females receiving 450 or 2000/1000 mg/kg/d. Marmosets receiving dosages of 50 mg/kg/d and above displayed several clinical signs which included emesis, diarrhoea, ptosis, occasional episodes of under- and overactivity, and excessive scratching activity. Skin reddening was observed during the first week of treatment in marmosets receiving 300 mg/kg/d. On the basis of the results obtained it can be concluded that the non-toxic doses of E-4868. Ts after 4-week oral administration in rats and marmoset monkeys were 100 and 25 mg/kg/d, respectively. PMID- 11413746 TI - Value of the dorsal cutaneous guinea pig model in selecting topical antiviral formulations for the treatment of recurrent herpes simplex type 1 disease. AB - Recurrent herpes simplex labialis represents a disease still difficult to treat, despite the availability of many established antiviral drugs used in clinical research since 30 years ago. Although differences between the human disease and that obtained in experimental animal suggest caution in predicting an effective clinical response from the experimental results, some of the animal models seem to be useful in optimising the topical formulation of single antiviral drugs. In the present work the dorsal cutaneous guinea pig model was used to compare 5 different topical antiviral formulations with clinical promise (active molecule: 5% w/w micronized aciclovir, CAS 59277-89-3), using both roll-on and lipstick application systems. The aim being to evaluate which vehicle (water, oil, low melting and high melting fatty base) and application system (roll-on, lipstick) enhances the skin penetration and the antiviral activity of the drug, after an experimental intradermal infection with Herpes simplex virus type 1 (HSV-1). As reference, a commercial formulation (5% aciclovir ointment) was used. The cumulative results of this study showed that the formulation A, containing 5% aciclovir in an aqueous base in a roll-on application system, has the better antiviral efficacy in reducing the severity of cutaneous lesions and the viral titer; among the lipsticks preparations, the formulation D, containing 5% aciclovir in a low melting fatty base, demonstrates a very strong antiviral activity, though slightly less than formulation A. This experimental work confirms the validity of the dorsal cutaneous guinea pig model as a rapid and efficient method to compare the antiviral efficacy of new formulations, with clinical promise, to optimise the topical formulation of the active antiviral drugs. PMID- 11413747 TI - [Anxiety and depression in the general population over 60 years old]. AB - BACKGROUND: Anxiety and depression lead to extensive utilization of the health service. Frequency and extent of symptoms of anxiety and depression were assessed in the elderly general population. PARTICIPANTS AND METHOD: In a random community sample of 622 elderly persons aged above 60 years (mean age 69.55 years; 56.1% women), the frequency of anxiety and depression was assessed by means of the Hospital Anxiety and Depression Scale. RESULTS: Probable anxiety was found in 7.6% of the elderly, probable depression in 27.5%. Negative affectivity (based on the total scale) was found in 31.7% of the elderly. Self-reported anxiety and depression symptoms correlated to a considerable extent (r = 0.69). There were higher rates of probable anxiety and depression in women than in men. However, a significant effect of the sex on anxiety and depression scores was not found. There was a significant effect of age, with higher depression rates with increasing age. Anxiety and depression correlate significantly positively with the fatigue syndrome and with subjective bodily complaints and significantly negatively with different aspects of quality of life. CONCLUSIONS: Physical complaints in the elderly cannot be directly traced back to age-correlated multimorbidity. Rather, elderly persons with nonspecific bodily complaints and fatigue should also be screened for anxiety and depression. PMID- 11413748 TI - [Endovascular stent graft support exclusion of an infrarenal aortic aneurysm using gadolinium as contrast medium in kidney insufficiency]. AB - HISTORY: A 64-year-old obese man with radiocontrast-induced nephropathy, which required temporary hemodialysis, was referred for treatment of an infrarenal abdominal aortic aneurysm detected incidentally. Comorbidities were obesity (BMI 44.1), non-insulin dependent diabetes mellitus, arterial hypertension, COPD, and renal failure with decompensated retention. INVESTIGATIONS: MRI demonstrated a 6.1 cm diameter infrarenal partially thrombosed aortic aneurysm which extended to the aortic bifurcation. Its morphology was favourable for endovascular treatment. TREATMENT AND COURSE: An aortobiiliac stent graft was inserted after bilateral arterial femoral cut-down. As a contrast agent we administered 0.5 molar gadoteridol solution only. The total volume used was 90 ml (0.375 mmol/kg body weight). The aneurysm was completely excluded. No further signs of renal deterioration occurred. CONCLUSION: Non-nephrotoxic gadolinium-containing contrast media allow minimal invasive exclusion of infrarenal aortic aneurysms with use of stent grafts in patients in whom conventional iodinated x-ray contrast agents are contraindicated. PMID- 11413749 TI - [Reversible blindness caused by an invasive prolactinoma]. AB - HISTORY: A 40-year-old man presented with severe headache, nausea and acute blindness. A CT-scan of the skull revealed a huge tumor along the basis of the skull. The patient was referred to our clinic for diagnostic and therapeutic evaluation. INVESTIGATIONS: MRT showed a large right-sided paramedian tumour displacing the brain stem with signs of increased intracranial pressure. Routine laboratory tests were normal except a normochromic anaemia. Endocrine tests demonstrated partial hypopituitarism with alteration of the somatotropic, gonadotropic and corticotropic axis and moderate hyperprolactinaemia. TREATMENT AND COURSE: On the day of admission a transnasal biopsy was taken. The preliminary histopathological diagnosis was a low differentiated carcinoma. Because of this diagnosis and because of the infiltrative tumour growth an operation was not performed but emergency irradiation was begun and dopamine agonist therapy was started because of hyperprolactinaemia. Several days later the final microscopic diagnosis of the transnasal biopsy specimen was reported to be an invasive prolactinoma. Under dopamine agonist therapy prolactin levels rose to a maximum of 6460 ng/ml to decline thereafter to normal values, and the visual disturbances recovered. After 5 weeks of therapy considerable shrinkage of the tumor was demonstrated by MRT. CONCLUSION: The differential diagnosis of acute visual deterioration caused by a large tumour along the basis of the skull includes an invasive prolactinoma. The diagnosis is made by demonstrating grossly elevated prolactin levels. To avoid falsely low prolactin measurements, caused by a hook-effect in the prolactin assay, serum dilution is mandatory in the diagnostic work-up. In the case of a prolactinoma medical treatment with dopamine receptor agonists is the therapy of choice because it causes rapid tumour shrinkage and symptomatic improvement in most patients, so that irradition of the tumour is not indicated. As dopamine agonist therapy is rapidly effective and well tolerated, it should be started even in case of doubt to lose no time until final diagnosis. PMID- 11413750 TI - [Amiodarone]. PMID- 11413751 TI - [Intracoronary ultrasound imaging]. PMID- 11413752 TI - [Therapy of severe pulmonary hypertension with prostacyclin and iloprost]. PMID- 11413753 TI - [Diagnosis of sprue]. PMID- 11413754 TI - The thymus in infancy and childhood. Embryologic, anatomic, and pathologic considerations. AB - Developmental anatomy and developmental and acquired disorders of the thymus are presented with an emphasis on those disorders that are relevant to infancy and childhood. Presented also is an attempt to correlate functional anatomy and pathologic and clinical findings. PMID- 11413755 TI - The thymus. Pediatric surgical aspects. AB - Since the original description of thymic death in an infant 400 years ago, the thymus has been recognized as an important structure to practitioners caring for infants and children. The source of many cysts, masses, and tumors in the neck and mediastinum, the thymus gland merits the pediatric surgeon's attention. The thymus is clearly an important lymphoid organ, the removal of which may be therapeutic in MG, but congenital absence leads to profound cell-mediated immunodeficiency. The immunologic sequelae of its neonatal extirpation remains obscure. It is apparent that further research is needed to clarify the functional role of the thymus gland in the developing immune system. Until better elucidated, a conservative approach to neonatal thymectomy may be justified. PMID- 11413756 TI - Imaging of the thymus. AB - Technologic advances in imaging, particularly CT and MR imaging, have revolutionized the ability to noninvasively image the thymus. Increasing knowledge of the normal and pathologic states of the thymus has been accompanied by an increasing ability to depict morphologic alterations in this gland with fine spatial resolution and can even provide functional information. This article discusses the role of various imaging modalities in identifying suspected thymic disease, which historically required surgery to visualize. The role of imaging is best understood with an appreciation of the embryology, anatomy, and pathology of the thymus. PMID- 11413757 TI - The thymus. Finally getting some respect. AB - The thymus gland is no longer a structure in the anterior mediastinum of questionable use in the body economy. It is the sole source of cells that control the immune response and provide protection from life-threatening pathogens. Its primary function is to seed the periphery with mature T cells. Because these cells can maintain their numbers through self-replication, thymic function essentially is completed within the first few years of life. In response to a lack of need for its services, the thymus thereupon undergoes involution. A demand for renewed thymopoiesis has become necessary as the T-cell compartment of the immune system must be replenished following massive depletion of T cells by disease (e.g., AIDS) or the demands of cytoreduction necessary to achieve successful bone marrow transplantation. In older patients with AIDS (> 20-30 years), re-expansion of the post-thymic population may be adequate to restore the patient to reasonable or even normal T-cell immunity, but when the cytoablation is as massive as seen with bone marrow transplant conditioning regimens, the thymus must contribute to the repopulation. If involution has proceeded too far, thymic output must be augmented. This augmentation can be accomplished by thymus transplantation. The techniques for successful thymus transplantation in adults, however, have yet to be perfected. The thymus can play a major role in determining the response to MHC products of others (i.e., nonself). Novel manipulations of the thymus are being used to achieve transplant tolerance. PMID- 11413758 TI - The thymus and myasthenia gravis. AB - Self acetylcholine receptor (AchR) is targeted by a wayward immune response in myasthenia gravis (MG). The current understanding of the pathogenesis of the AChR directed immune response is reviewed. Furthermore, the thymus is suspected of initiating and perpetuating the disease process in the majority of patients; its role as a central and peripheral lymphoid organ in MG is discussed. MG seems to result from a failure of (1) establishing tolerance to the AChR and (2) regulating the immune response. PMID- 11413759 TI - Medical therapies in myasthenia gravis. AB - Forty years ago, a patient with MG probably had a fifty-fifty chance of surviving a myasthenic crisis, defined as the need for mechanical ventilatory support. Approximately 16% of all patients experience a crisis, a figure that has not changed appreciably since then. Progressive weakness, oropharyngeal symptoms, refractoriness to anticholinesterase medication, intercurrent infection, and invasive procedures including needle biopsies of thymic gland masses, and reactions to contrast agents used in the performance of CT of the chest have been implicated in the development of crisis. It is now standard practice to treat severe crisis in an intensive care unit. The ready availability of intensive care in most hospitals belies the fall in the mortality of myasthenic crisis to 6% over the past several decades. Crisis is a temporary exacerbation, regardless of the proximate cause, and the goal is to keep the patient alive until it subsides, usually in 2 weeks. In the past, edrophonium was used to differentiate myasthenic crisis from cholinergic crisis, but that is now moot because withdrawal of cholinesterase medication is necessary for improvement in both situations. The underlying immunologic derangements in myasthenic crisis are not well understood, but there is a rapidly fatal antibody-mediated syndrome that bears resemblance to crisis and is associated with inflammation and necrosis of the end-plate region. PMID- 11413760 TI - Anesthesia and critical care of thymectomy for myasthenia gravis. AB - Myasthenia gravis is an autoimmune disease resulting from the production of antibodies against the ACh receptors of the neuromuscular synapse. The thymus gland is involved in the autosensitization process, and the disease frequently is associated with thymic morphologic abnormalities. There is a consensus that all adults with generalized MG should have a thymectomy. This recommendation has been propagated by the safety of the procedure and excellent outcome. Removal of as much thymic tissue as possible (anterior mediastinal exenteration) by transsternal approach is the logical goal of thymectomy in the treatment of MG. Transcervical approach and VATS, however, are less invasive and have been used in patients who have MG unaccompanied by thymoma. Optimization of the condition of the myasthenic patients can markedly decrease the risk of surgery and improve the outcome. Two techniques have been recommended for general anesthesia in the myasthenic patient. Because of the unpredictable response to succinylcholine and the marked sensitivity to nondepolarizing muscle relaxants, some anesthesiologists avoid muscle relaxants and depend on deep inhalational anesthesia, such as halothane, isoflurane, or sevoflurane, for tracheal intubation and maintenance of anesthesia. Others, however, use a balanced technique of anesthesia that includes the use of carefully titrated muscle relaxants. The most important preoperative factor predicting the need for postoperative mechanical ventilation is the severity of bulbar involvement (Ossermann group 3 and 4), usually indicated by significant dysphagia and dysarthria associated with borderline respiratory dysfunction. Thymectomy benefits nearly 96% of patients: 46% develop complete remission, 50% are asymptomatic or improve on therapy, and 4% remain the same. The time from diagnosis to surgery is shorter than 8 months, and mild or moderate myasthenic symptoms are the main prognostic factors that predict the best outcome after thymectomy. PMID- 11413761 TI - Transcervical thymectomy for myasthenia gravis. AB - The authors of this article contend that the transcervical approach for thymectomy allows the safe and complete removal of the thymus and [table: see text] provides equivalent benefit to the patients with regard to opportunity for clinical remission or freedom from progression of the symptoms of MG. The low morbidity and short hospitalization after transcervical thymectomy represent minimal barriers and allow increased willingness of the neurologist to refer a patient for surgical therapy and increased acceptance of the patient towards the recommendations. The authors believe that an early, safe, and complete thymectomy offers all the benefits of surgical removal of the thymus to a patient with MG with minimal risk for morbidity and postoperative pain. PMID- 11413762 TI - Extended trans-sternal thymectomy for myasthenia gravis. AB - Otto reported thymectomy under transverse sternotomy. It currently is not a widely used procedure, however. Jaretzki graded the completeness of removal of thymic tissue as follows: maximal thymectomy 98% to 100%, extended thymectomy 85% to 95%, modified transcervical thymectomy (Cooper) 75% to 80%, VATS thymectomy 80% to 85%, simple trans-sternal thymectomy 70% to 75%, and simple transcervical thymectomy 40% to 50%. The results of each procedure do not always reflect the completeness of removal. The tables show that the most widely accepted procedure is the extended thymectomy, and the results of it are prominent and stable. Why do the results of the maximal thymectomy, however, not exceed those of the extended thymectomy? It is supposed that quantitative increase of removed thymic tissues in the maximal thymectomy might be minimal. PMID- 11413763 TI - Video-assisted thoracoscopy thymectomy for myasthenia gravis. AB - Over the past 8 years, the technique of video-assisted thoracoscopic surgery (VATS) thymectomy has continued to evolve. Although the procedure has become better defined and greater experience has been gained, numerous issues still exist. Whether it is best performed through a left sided, right sided, or as a bilateral approach with or without a cervical incision, is not clear. Equivalence of outcomes compared with more standard approaches to thymectomy is still an issue. Experience still is relatively limited to a few centers, and follow-up still is relatively short. In the author's own experience, it seems that availability of the less invasive approach has allowed thymectomy to be performed earlier in the course of the disease especially in young female patients in whom cosmesis is an important issue. PMID- 11413764 TI - Histologic typing of thymoma according to the new World Health Organization classification. AB - The new WHO classification scheme provides a set of simple and easily reproducible morphologic criteria for an accurate and consistent categorization of thymic epithelial tumors. This scheme should facilitate interobserver reproducibility and lead to a more precise assessment of prognosis when used in combination with the staging system. PMID- 11413765 TI - Surgery for thymoma. AB - Although thymoma is an uncommon tumor, it represents the most frequently encountered tumor of the anterior mediastinum. These tumors represent an interesting and even peculiar group of lesions by virtue of their association with paraneoplastic disorders, their relatively indolent course, and their predisposition for local recurrence. The initial treatment of choice for patients with thymoma that do not present with unresectable local or diffuse metastatic disease is complete surgical resection. The goals of surgery are complete excision of the lesion with total thymectomy and complete exploration to rule out the presence of noncontiguous disease that may be resectable. Often, complete resection may require the resection of surrounding involved structures including pericardium, pleura, lung, and even major vascular structures. Some authors have suggested VATS or VATS-assisted techniques for small thymomas. Capsular invasion, however, often can be subtle, and the completeness of resection is of prime importance in countless studies. With recurrences appearing up to 5 and even 10 years postoperatively, time will tell if these minimally invasive techniques are comparable with current standard approaches. Multiple studies have failed to determine conclusively the role of induction chemotherapy and adjuvant radiation. Prospective multi-institutional trials are required to elucidate further the role of such therapies in these rare tumors. In the interim, the authors continue to recommend postoperative radiation for all patients undergoing resection with the exception of stage I patients. Some promising reports on response to chemotherapy have led them to develop an induction chemotherapy protocol for patients with clinically advanced disease. PMID- 11413766 TI - Reoperation on the thymus. A critique. AB - Reoperation on the thymus, including even multiple re-resection, is a useful modality in carefully selected patients for recurrent thymoma or for incompletely removed thymus. In the case of MG, reoperation is used for completion of an otherwise incomplete thymectomy, usually by the transcervical method, although it also has been used after primary sternotomy. Interestingly enough, no instances of reoperation after primary VATS thymectomy have been reported yet. Indeed, VATS has been used successfully for reoperation after previous transcervical and trans sternal thymectomies. In the case of thymoma, there have been an increasing number of reports of reoperation, including multiple procedures, in many cases. It is important to interpret the results of reoperation in the context of the biologic behavior of the thymus tumor. Thus far, no formal guidelines have been established for the follow-up of these patients. It is believed that all cases of thymectomy with or without thymoma and with or without MG should be followed for the lifetime of the patient with periodic examinations and imaging. PMID- 11413767 TI - Chemotherapy of thymomas and thymic carcinomas. AB - Thymomas are chemosensitive tumors with overall response rates of about 70% to various chemotherapy regimens. A fraction of patients (up to 25%-30%) will obtain a CR after chemotherapy. These results justify the use of chemotherapy in a multimodality fashion for the treatment of patients with advanced tumors. With radiotherapy added to chemotherapy, if necessary and if technically feasible, inoperable tumors may become resectable, leading to excellent long-term survival. On the other hand, thymic carcinomas are more refractory to chemotherapy, and their prognoses remain poor. PMID- 11413768 TI - The radiotherapeutic management of invasive thymomas. AB - The role of radiation therapy has been well established for selected patients with thymoma. Among patients with stage I tumors that have undergone a complete surgical excision, there is no indication for postoperative RT. There is also no need for postoperative RT for patients with completely excised stage II tumors when there is no transgression of the tumor through the thymic capsule. If there is transgression of the tumor through the capsule, postoperative RT is recommended even in the presence of complete surgical resection. For totally or partially resected stage III tumors, postoperative RT, to a total dose in excess of 50.4 Gy, is recommended. CT scan-based treatment planning is recommended for all of these indications. The role of preoperative RT is currently less certain. For bulky, unresectable tumors, preoperative chemotherapy and postoperative RT seem to be a promising trimodality approach. There also will be a need for preoperative RT among partial responders. PMID- 11413769 TI - Tuberculosis: the beginning of the end? PMID- 11413770 TI - Effect of albumin-furosemide mixtures on response to furosemide in cirrhotic patients with ascites. PMID- 11413771 TI - From ETOH to FAB: the medicalization of therapy for pit viper envenomation. PMID- 11413772 TI - Dementia in the elderly: the "silent epidemic" no more. PMID- 11413773 TI - Farming for spare body parts: silk purse or sow's ear. PMID- 11413774 TI - Silicone breast implants: lessons from a saga. AB - In the following eleven lessons learned from the silicone breast implant saga in the United States are listed. Some Lessons From The Saga Of Silicone Breast Implants In The U.S. 1. The intrinsic differences between science and the law mean that there will continue to be tension at the interface between the two. 2. Weak scientific evidence makes for weak scientific, clinical and legal judgments. 3. Health policy can influence the weighing of evidence as did the FDA ban on gel filled silicone breast implants in 1992. 4. As the probative value of scientific evidence decreases (that is, the quality and relevance of the evidence) the courts have increasing difficulty in evaluation. 5. Weak scientific evidence along with poor legal judgments can distort clinical understanding and result in harm to patients. 6. Widely publicized speculation and litigation can obscure less dramatic but frequent and serious complications. 7. Clinicans, toxicologists, industry and regulators should maintain surveillance of medical devices, in representative groups if not in all recipients of the devices. 8. The precise identity and composition of each medical device implanted into a person should be recorded both by the manufacturer and in the health care record. 9. The factual basis for informed consent should be vetted by a disinterested party. 10. Patients, clinicians and industry all have a strong interest in assuring the safety of medical devices through regulation. 11. The criteria and process for evaluating the admissibility of scientific evidence in legal proceedings have been improved but universal adoption of the improvements has not been accomplished and further changes may be necessary. PMID- 11413775 TI - The Jeremiah Metzger Lecture. Hypercoagulable states: challenges and opportunities. AB - Thromboregulatory physiology is essentially a function of the blood vessel wall. Constitutive endothelial cell activities maintain blood fluidity by down regulating the initiation as well as, the propagation of blood coagulation. The major systems involved include: the Protein C, TFPI, plasmin generating and antithrombin pathways, all of which are focused on the cell membrane. Altered regulation of these endothelial functions forms the basis of the pathophysiologic events associated with the inherited primary hypercoagulable states. Secondary hypercoagulable syndromes occurring in various clinical states with conversion to a vascular thrombogenic phenotype reflect non constitutive activated endothelial cell functions with concomitant down regulation of the constitutive anticoagulant surface activity. So called idiopathic clinical thrombosis in most circumstances represents multi hit events in which specific genetic abnormalities or polymorphisms together with specific acquired alterations in geographically distinct endothelial cell beds culminate in a recognizable coagulation phenotype. PMID- 11413776 TI - Theodore E. Woodward Award. Do ANCA cause small vessel vasculitis? AB - ANCA may be a pathogenetic force, but to date, support for this contention remains indirect. Active immunization with antigen or passive transfer of ANCA has not reproduced small vessel vasculitis (SVV). It is more than likely that if ANCA are pathogenetic, they are involved as one of many simultaneously occurring mechanisms acting in concert with other synergistic inflammatory mediators of disease. These include not only environmental factors such as infection or environmental toxins such as silica, but also genetic factors that are only now being described. The paradigm for this autoimmune process must include several events that occur simultaneously or sequentially, including ANCA, leukocyte activation and injured endothelium. PMID- 11413777 TI - The protean nature of Whipple's disease includes multiorgan arteriopathy. AB - Knowledge about the arterial abnormalities in Whipple's disease can be useful for our better understanding of both Whipple's disease and the more general question of pathogenesis of atherosclerosis. There are several notable morphological features of Whipple's arteriopathy. First, it appears to involve primarily arteries one millimeter or less in diameter. Second, there is very little evidence of inflammation accompanying invasion of any or all three layers of the walls of affected arteries, and there is almost no evidence of local attraction of platelets to these sites of arterial injury. Third, the nature of arterial injury appears to be one of slow progression. The few sites of actual arteritis are most likely attributable to some other coinciding microbial organism not yet identified. Although the arteriopathy in Whipple's disease is seen mainly in small arteries (the aorta is a notable exception), their significance can be illustrated by consideration of this fact as it applies to the coronary circulation (and probably the arteries of all other organs). In the heart these small arteries comprise almost the entire collateral circulation, the principal blood supply to each component of the conduction system, and most pragmatically, these small arteries represent the terminal distribution of every larger epicardial artery. Small arteries are important. The "cardiomyopathy" so often a feature of Whipple's disease (very much including his original case) is most logically attributable to recurring bouts of focal ischemia and subsequent focal fibrosis ending in myocardial incompetence. However, direct bacillary invasion of cardiac myocytes (22) also occurs. In lamina propria of jejunum, there is also arteriopathy, as there is in brain, lung, kidney, spleen, liver, gall bladder, rectum, stomach, lymph nodes and testis. It is likely that no organ in the body is spared. There is growing evidence that a wide variety of chronic infections (occurring concomitantly or sequentially) may participate in the early pathogenesis of human arterial disease, including atherosclerosis. Given that the coronary plaque represents the cumulative end result of countless earlier injuries and responses, the plaque is not the site to seek evidence of initial pathogenesis although understanding the behavior of coronary plaques is eventually of considerable clinical importance. In the context of original events in pathogenesis, the Whipple bacillus now deserves inclusion in the "total pathogen burden" concept, as it relates not only to coronary disease but to all aspects of atherosclerosis and even other forms of arteriopathy. PMID- 11413778 TI - Microbes, inflammation and atherosclerosis: will old pathology lessons guide new therapies? AB - Although attractive, the microbial pathogenesis theory for atherosclerosis remains unproven. Over the last century, microbiologists have invoked fulfillment of Koch's postulates to determine pathogen causality. Certainly a multifactorial disease process such as atherosclerosis unlikely will be due to a single microbial agent, an agent when transferred to another host, will always induce atherosclerosis. Conflicting epidemiological data also do not support a single causative agent. However, as presented here, considerable in vitro, animal, and human epidemiological data support the plausibility that infectious agents can promote a proinflammatory, procoagulant and proatherogenic environment in the vessel wall. Microbial genes and molecules can catalyze these processes and foil normal cellular events. But, must intact microbes enter the vessel wall or can microbial molecules incite immune responses from afar? A new focus on pathogen induced auto-immunity toward vasculature has been presented. For example, microbes contain molecules that mimic host cellular components (55). An immune response to a pathogen may cross react with vessel wall cellular structures. This immune response enhanced by infection may lead to high levels of cross reacting auto-antibodies or auto-aggressive T-cells. Epstein has championed the concept of pathogen burden in support of this auto-immune theory (56). Individuals infected with multiple pathogens such as HSV-1, HSV-2, CMV, Helicobacter pylori, and Hepatitis A, have high C-reative protein levels (markers of inflammation) and the greatest relative risk for coronary artery disease (57). Thus, pathogens might contribute to the atherosclerotic process by promoting inflammatory responses. It is this author's view that microbes and inflammation do play a role in the pathogenesis of atherosclerosis (58). Infection may contribute to the process promoting vessel wall injury initiated by oxidized lipids, smoking derived oxidants, hypertensive shear or diabetes glyoxidized molecules. Inflammation and immune reactions in response to infection can exacerbate and act synergistically with all of the aforementioned vasculotoxic moieties. Continued investigations in the 21st century will determine if vaccines, antibiotics, anti-inflammatory agents or immunosuppressants will alter the picture the early 19th century pathologists observed under their monocular microscopes. PMID- 11413779 TI - Cutaneous manifestations of infectious diseases: approach to the patient with fever and rash. PMID- 11413780 TI - TGF-beta: a crucial component of the pathogenesis of diabetic nephropathy. AB - In summary, metabolic, hemodynamic, and genetic factors are all important in the development and progression of diabetic nephropathy (33). Recent studies using cell culture techniques and experimental animal models have provided important insight into the role of hyperglycemia in this disease. The nature of the factors directly arising as a consequence of hyperglycemia and the steps involved in diabetic complications are not completely understood. The characteristic lesions of diabetic nephropathy may be intimately related to the effects of high ambient glucose on intracellular signaling events, various growth factors/cytokines, and nonenzymatic glycation of proteins (36). The growth factor TGF-beta has emerged as a key participant in the cascade of events which leads to kidney sclerosis. Increased TGF-beta expression in the kidney in diabetes mellitus mediates the renal actions of high ambient glucose to promote cellular hypertrophy and stimulate extracellular matrix biosynthesis. Neutralizing the actions of TGF-beta with highly specific monoclonal antibodies or with application of antisense technology can effectively prevent the induction of the kidney lesions of diabetes in mice independent of any changes in blood glucose levels. Such maneuvers are crucial for establishing proof-of-concept and Koch's postulates (17), but they are still far removed from clinical applicability. Nevertheless, it is hoped that further studies to elucidate the efficacy of novel interventions to intercept the activity of the renal TGF-beta system will prove useful for effectively halting the progression of diabetic nephropathy. PMID- 11413781 TI - Blueprint for discovery in academic medicine: plans, process and outcomes. AB - By the end of the decade, we had fully implemented most of the recommendations of the Molinoff Report. Our programmatic analysis is summarized in Table 11. While the space needs identified in the Molinoff Report were met by BRB I, II, and III (289,000 nsf as compared [table: see text] to 276,000 nsf as planned), it was possible to provide additional, somewhat unanticipated, research space (111,000 nsf) prior to the end of the decade. The faculty has now developed a research plan for the next decade. It is also important to emphasize that the total faculty grew by 41% [table: see text] over the decade and most of that growth occurred with faculty spending a substantial part of their time in clinical practice. Hence, the dramatic improvement in research funding of over 200% was due largely to the enhanced productivity of our faculty. By taking an organized planning approach deeply seated in the faculty, consistent with Trustee directives and with measurable outcomes, we were successful in growing the research programs within the School of Medicine of the University of Pennsylvania. We believe this particular approach, taken with a focus on multidisciplinary research, [table: see text] was the right one for the 1990s. In the final analysis, it is abundantly clear that outstanding faculty, working in an exciting supportive environment, was the most important factor for success. We are not certain what the right approach will be for the future. Clearly, with the important advances in genomics and information technology, the importance of the team, even if a virtual one world-wide, cannot be overstated. While research is only one mission of the School of Medicine, clearly, our visible success in research played an important role in the overall improvement in the School of Medicine as measured by others. For example, the ranking of the School of Medicine by U.S. News & World Report, perhaps the most widely used ranking by the lay press, went from 10th to 3rd behind only Harvard and Johns Hopkins during the period of the 90s (Table 12). PMID- 11413782 TI - William Osler Abbott: his double lumen tube. AB - Miller as head of the GI Section, Abbott's Chief and the individual who actually constructed the original double lumen tube was logically lead author of their first paper describing a technique for intubation of the human intestine. Nonetheless, originally most people associated only Abbott's name with the tube since he arranged for its manufacture, perfected the technique by which it was used, described its use in the diagnosis and treatment of intestinal obstruction and personally carried out over 500 intubations. Three months after his death Shields Warren wrote "Not a day goes by here in the Hospital that a life is not saved by the Abbott tube" (16). Were it not for Abbott's correction of the Pilling company's first brochure the Miller Abbott tube might have been called the Abbott tube or at the very least the Abbott Miller Tube. Such a designation would not have been inappropriate when one also considers that Pete Abbott's use of the double lumen tube in studies of the gastrointestinal tract and the diagnosis and treatment of intestinal obstruction was undoubtedly a major factor in his untimely death. PMID- 11413783 TI - Feedback control of pancreatic exocrine secretion: its relationship to the management of the abdominal pain associated with chronic pancreatitis. PMID- 11413784 TI - Use of recombinant angiostatin to prevent retinal neovascularization. PMID- 11413785 TI - The Gordon Wilson Lecture: viruses and human disease. AB - In many ways, Ebola virus infection provides a model for understanding the toxicity of viruses and their causal role in human disease. The highly aggressive course of Ebola virus infection provides a model for understanding the molecular mechanisms of viral cytotoxicity. In addition, the use of animal models and definition of immune correlates, which lead to protection, may provide lessons that are applicable to other viral infections. Perhaps the greatest challenge facing biomedical science today is the containment of the human immunodeficiency virus, the causative agent of AIDS. In many ways the critical obstacles to the development of a vaccine for HIV are similar to those observed with Ebola virus infection. Because the reservoir of infection is not known and human-to-human spread has been documented, vaccines may provide the best opportunity to contain and limit the spread of infection worldwide. Similar to Ebola virus, there are few convincing examples of immune resistance of HIV infection. In addition, it has been difficult to identify broadly neutralizing antibodies that can prevent infection in vitro or in vivo. In defining immune correlates, relevant animal models, and mechanisms of cytotoxicity, it is hoped that similar efforts may lead to effective vaccines for other infectious diseases. In this way, Ebola virus infection provides a useful paradigm for understanding the genetic determinants of viral disease and in facilitating the development of treatments and prevention of viral infections. PMID- 11413786 TI - Primary and secondary prevention of heart disease: can we curb the global epidemic? PMID- 11413787 TI - Pathogenesis of cystic fibrosis airways disease. PMID- 11413788 TI - Memorial. Sven Martin Gundersen 1904-1998. PMID- 11413789 TI - Memorial. Thomas William Mattingly 1907-1999. PMID- 11413790 TI - Memorial. Sir John W. H. Butterfield 1920-2000. PMID- 11413793 TI - Memorial. Jay P. Sanford 1928-1996. PMID- 11413795 TI - [Obesity: a weighty problem for cardiovascular medicine]. PMID- 11413796 TI - [Obesity and cardiovascular risk]. AB - PROBANDS AND METHOD: In the Prospective Cardiovascular Munster (PROCAM) Study, 16,288 men aged 40.5 +/- 11.3 years (mean +/- SD) and 7,328 women aged 35.8 +/- 12.3 years were enrolled between 1979 and 1991. RESULTS: Mean body mass indices (BMI) were 25.6 +/- 3.3 kg/m2 and 23.8 +/- 4.1 kg/m2 in men and women, respectively. There was a graded positive interaction in both men and women between BMI and age-standardized levels of serum total cholesterol, low density lipoprotein cholesterol, triglycerides, fasting blood glucose, uric acid, and blood pressure (both systolic and diastolic). High density lipoprotein cholesterol tended to decrease with increases in BMI in both sexes. In the subgroup of male participants aged 40-65 years without a prior history of myocardial infarction or stroke in an 8-year follow-up 258 major coronary events were observed. Although incidences of major coronary events increased with increase in relative body weight in a multiple logistic function analysis BMI did not independently contribute to cardiovascular risk, indicating that the effect of overweight on coronary heart disease is mediated via other risk factors. PMID- 11413797 TI - [Molecular principles of obesity]. AB - BACKGROUND: The deposition of excess amounts of energy in adipose tissue is enhanced by high-fat diets and lack of physical activity. Furthermore, the existence of a specific genetic predisposition towards the development of obesity becomes evident by marked interindividual differences in the response to caloric oversupply. GENETIC DEFECTS AND HORMONES: In recent years, numerous genes and genetic defects with importance for human obesity were identified, especially through studies in animal models. The adipocyte-derived hormone leptin and its hypothalamic receptor play a premier role, as they interact with a network of proteins and neuropeptides within the regulation of food intake and energy expenditure. CONCLUSION: The search for the key molecular mechanisms in the pathogenesis of obesity will not only improve our understanding of energy metabolism, but may ultimately also lead to the development of new treatment strategies for obese patients. PMID- 11413798 TI - [Psychological factors in development and treatment of obesity]. AB - BACKGROUND: The prevalence of obesity has distinctly increased in almost all industrial countries in the past decades and is considered today to be one of the most important health problems. In Germany, between 12 and 18% of the adult population is obese. Besides the known risk factors such as arterial hypertension, lipometabolism disorders and type-2 diabetes mellitus, obesity is connected to severe psychosocial burdens. The genesis of obesity is multifactorial, besides genetic factors environmental ones such as an increase in hypercaloric food and at the same time a reduction of physical activity play a decisive role. PSYCHOGENESIS: Whereas earlier mainly psychic factors were discussed in the genesis of obesity, and here especially the question as to which personality variables are most frequently associated with obesity, the discussion in the past decades has increasingly shifted towards psychosocial consequences. It does, however, seem justified to adhere to a "psychogenesis" of obesity in a subgroup of obese people. Psychosomatic/psychiatric disorders lead to a change in eating behavior and physical exercise, the result being a sustained positive energy balance with overweight and obesity. PSYCHOSOCIAL PROBLEMS AND TREATMENT CONCEPTS: This paper describes the psychosocial problems of obese people with an emphasis on the question of psychiatric co-morbidity in general and eating disorders in particular. The results of mainly behavioral therapeutically oriented treatment concepts are presented and school-overlapping concepts discussed. PMID- 11413799 TI - [Cardiovascular changes in obesity]. AB - BACKGROUND: Obesity is associated with a number of adaptations in the cardiovascular system like arterial hypertension, a hypervolemic circulation or left ventricular hypertrophy. PATHOBIOCHEMICAL MECHANISMS: The specific mechanisms underlying these sequelae of obesity are not fully understood. Metabolic factors like insulin-resistance or the elevated activation of the sympathetic and the renin-angiotensin-aldosteron system have effects on a number of vascular and cardiac parameters. In addition, elevated serum levels of proinflammatoric parameters like C-reactive protein, interleukin-6 and TNF-alpha are found in patients with obesity. The consequence is a drastic increase of cardiovascular mortality. CONCLUSION: Depending on the fat distribution and the relation between the increase of body fat mass and fat-free mass different effects on heart and vessels are reported. The most important obesity-associated implications on the cardiovascular system are discussed below. PMID- 11413800 TI - [Non-drug therapy of obesity]. AB - BACKGROUND: The current epidemic of obesity is primarily a consequence of the modern lifestyle characterized by a hypercaloric, high-fat diet and lack of physical activity. INTEGRATED TREATMENT CONCEPT: In order to establish a treatment strategy oriented at the pathogenesis of the disorder it is necessary to strive for a long-term change in lifestyle. The modern best-practice treatment of obesity includes a healthy, low-fat diet, an increase in physical activity and behavior modification. This integrated treatment concept aims at decreasing body weight by 5-19% and at obtaining long-term weight management. PMID- 11413801 TI - [Treatment of hypertension in obesity]. AB - BACKGROUND: Hypertension and obesity are common medical conditions independently associated with increased cardiovascular risk. Many large epidemiological studies have demonstrated associations between body mass index and blood pressure, and there is evidence to suggest, that obesity is a causal factor in the development of hypertension in obese subjects. Weight Reduction and maintenance is an essential first step in the treatment of obesity-associated hypertension. Weight reduction may be achieved by behavior modification, diet, and exercise or by the use of anti-obesity medication. However, the long-term outcomes of weight management programs for obesity are generally poor, and most hypertensive patients will require antihypertensive drug therapy. PATHOPHYSIOLOGY: Obese hypertensive patients often have metabolic abnormalities known to be exacerbated by commonly used antihypertensive agents but also obesity per se is often associated with endorgan damage including left ventricular hypertrophy, glomerular hyperfiltration and microalbuminuria, congestive heart failure or sudden cardiac death. Furthermore they have revealed volume expansion, increased cardiac output, and lower total peripheral resistance than lean patients. Hypertension in obese patients appears to be related to both increased sympathetic nervous system activity and activation of the renin-angiotensin system. Where antihypertensive therapy is necessary, the aim should be to use agents based on the hemodynamic and metabolic background and that have benefits beyond blood pressure lowering and improve the conditions most commonly linked with obesity-associated hypertension, such as hyperlipidaemia, Type II diabetes, left ventricular hypertrophy, coronary artery disease, or congestive heart failure. PHARMACOTHERAPY: Based on their favorable metabolic profiles, it would appear that ACE inhibitors, angiotensin receptor blockers, calcium channel blockers, moxonidine and alpha-blockers can lower blood pressure without worsening the metabolic abnormalities, that is just one aspect of the problem. Yet, most guidelines fail to provide specific advice on the pharmacological management of hypertension in obese patients. This may be due to the fact that there are currently no studies that have addressed the efficacy of specific antihypertensive agents in reducing mortality in obese-hypertensive patients. This paper reviews the theoretical reasons for the differential use of the major classes of antihypertensive agents in the pharmacological management of obesity related hypertension and also considers the potential role of anti-obesity agents. PMID- 11413802 TI - [Anesthesia in extreme obesity]. AB - BACKGROUND: Approximately 1-2% of all anesthetized patients are morbidly obese (body mass index > 35 kg/m2). The perioperative mortality is significantly elevated (up to 20%) compared with lean patients. Morbidly obese patients are at high risk for cardiopulmonary dysfunction. Difficult airway management is reported in 13-20% of obese patients. Hypoxia is often observed due to faster desaturation during induction of anesthesia. After surgery, patients are endangered by a high incidence of obstructive sleep apnea syndrome (50%), pulmonary atelectasis (5%) and acute pulmonary embolism (5-12%). ANESTHESIA: Individualized perioperative management is required based on preoperative history and physical examination. Modern anesthetic drugs (desfluran, sevoflurane or propofol, and remifentanil, respectively) allow rapid recovery and early postoperative mobilization. Adequate monitoring, e.g. by an intraarterial blood pressure monitoring and repetitive blood gas analyses, improves patient safety prior the onset of complications. POSTOPERATIVE MANAGEMENT: Postoperative admission on an intensive care unit of morbidly obese patients is based upon concomitant diseases and surgical requirements. The main reason for admission is an inadequate pulmonary gas exchange. This interdisciplinary approach will reduce the risk of anesthesia and avoid complications in morbidly obese patients. PMID- 11413803 TI - [Syncope of unknown origin in heart failure: which diagnostic possibilities should be considered today?]. AB - In patients with congestive heart failure, unexplained syncope is often due to ventricular arrhythmias and associated with a poor prognosis. Electrophysiological studies should be considered early in the work-up of syncope. Implantation of a defibrillator might become necessary in many patients with syncope and heart failure. PMID- 11413804 TI - [Deficiencies in health care. Health services research--bridge between medical quality assurance and financing crisis]. PMID- 11413805 TI - [The French Association for Endocrine Surgery]. PMID- 11413806 TI - [Endocrine tumors of the duodenum and pancreas: "prerequisites", imaging, and intraoperative detection]. AB - Non-secreting pancreatic neuroendocrine tumors and diarrheogenic tumors are usually bulky and easy to recognize thanks to somatostatin-receptor scintigraphy and hormonal dosages. The real difficulty concerns insulinomas and particularly gastrinomas, which are found at the level of the duodenum and can be very small. Diagnosis is based on biological examinations, and the search for MEN I must be done systematically. Their localization is based on two essential and noninvasive exams: echoendoscopy and somatostatin-receptor imaging, though the surgeon's skill in the operative field, aided by an intraoperioperative echography, is indispensable. Surgical treatment of insulinomas gives excellent results. However, for sporadic gastrinomas, there was only a 34% recovery rate, and still less for MEN I gastrinomas. PMID- 11413807 TI - [Risk factors for Crohn's disease relapse after treatment of intestinal stenosis]. AB - BACKGROUND: Postoperative recurrences of Crohn's disease (CD) has been widely investigated in previous studies. Nevertheless, the risk factors for CD recurrence in patients presenting with intestinal stenosis are not clearly identified. PATIENTS AND METHOD: Thirty consecutive patients out of a cohort of 134 patients with CD presented with an intestinal stenosis diagnosed between 1995 and 1999. Epidemiological, clinical, and therapeutic data were carefully recorded. A univariate analysis followed by stepwise descending discriminant analysis was performed. RESULTS: Sixteen patients (53%) underwent surgery, six received steroid therapy, eight were given antibiotics, and seven took immunosuppressive drugs. The mean follow-up after medical or surgical management was 2.8 years. Thirteen patients (43%) had CD relapse during the follow-up. Considering the univariate analysis, the existence of a previous appendectomy and the persistence of tobacco consumption were significantly associated with the risk of CD relapse during the follow-up. The stepwise descending discriminant analysis identified three independant factors: tobacco consumption (p = 0.007), previous appendicectomy (p = 0.04) and duration of the follow-up (p = 0.02). CONCLUSION: The CD relapse after the management of small bowel stenosis occurred in 43% of the patients within a mean follow-up of 2.8 years. The significant factors associated with the risk of CD relapse were tobacco consumption, previous appendectomy and duration of the follow-up. PMID- 11413808 TI - [Para-aortic lymph node involvement in gastric adenocarcinoma]. AB - AIMS: To determine the significance of superextended lymphadenectomy (D4) in patients with gastric cancer. The incidence of para-aortic lymph node metastases (N4) was analysed as well as its relationship to the site of the tumour. PATIENTS AND METHODS: The frequency of para-aortic lymph node metastases was assessed in 110 patients who underwent gastrectomy with D4 lymphadenectomy during the period from June 1988 to October 1999; five patients with plastic linitis and three with carcinoma of the gastric stump were excluded from the study. RESULTS: The postoperative mortality rate was 2.7% (n = 3) and the postoperative morbidity rate was 29.1% (n = 32). In our experience the most frequent postoperative complications were pancreatic fistulas (7.3%) and respiratory complications (6.4%). Among the 110 patients, the total number of dissected nodes was 5245 and the mean number of dissected nodes per case was 47.7. The total number of retrieved lymph nodes from the para-aortic station level was 639, with a mean number of 5.8 per patient. N4 nodal involvement was found in 20 (18.2%) out of 110 patients: 12 (33%) patients with a carcinoma located in the proximal third, two (6%) with a tumour located in the middle third and six (15%) with a carcinoma of the distal third of the stomach. CONCLUSION: The presence of para-aortic lymph node involvement in 18.2% of the patients suggests that D4 lymphadenectomy should be considered in the curative surgical treatment of advanced gastric cancer, especially if located in the proximal third of the stomach (N4 + in 33% of the patients). PMID- 11413809 TI - [Free-flap reconstruction of locally advanced soft tissue sarcomas]. AB - STUDY AIM: The aim of this retrospective study was to assess the advantages of free-lap reconstruction in locally advanced soft tissue sarcomas. PATIENTS AND METHOD: From October 1997 to October 2000, among 256 procedures for sarcomas, 25 were associated with free-lap reconstruction. There were 16 women and nine men (mean age: 45 years). The tumor was located on inferior limbs (n = 15), superior limbs (n = 3) and trunk (n = 7), had a mean size of 12, 10 and 12 cm respectively, and was multifocal (n = 6), or recurrent (n = 10). Ten patients received a neoadjuvant chemotherapy. Tumoral excision was associated with a complementary procedure in 10 patients. Reconstruction was performed with free laps of the latissimus dorsi (n = 21) and transverse rectus abdominis myocutaneous (TRAM) (n = 4). Early postoperative radiotherapy was associated in 15 patients. RESULTS: Median duration of the procedure was 6.5 hours. There were no postoperative deaths. Two free-laps necrotized and another free-lap was constructed. Tumoral excision was Ro (n = 20), R1 (n = 4) and R2 (n = 1). With a median follow-up of 20 months, there were no local recurrences in patients Ro. Ten patients developed pulmonary metastases and five of them died. CONCLUSION: With free-lap reconstruction, tumoral excision was larger, amputation of the limb was avoided in 14 patients, and early postoperative radiotherapy was possible in 15 patients. Multidisciplinary management and collaboration of the oncologist and plastic surgeon are associated with a better prognosis of locally advanced soft tissue sarcomas. PMID- 11413810 TI - [Videoendoscopic approach for parathyroid adenomas: results of a prospective study of 100 patients]. AB - STUDY AIM: The aim of this prospective study was to report the results of 100 consecutive video-endoscopic parathyroidectomies in patients suspected of having a single adenoma. PATIENTS AND METHOD: From March 1997 to September 2000, 80 females and 20 males (mean age: 49.5 years) were operated on. They were selected on the basis of the following criteria: preoperative imaging in favour of a single adenoma, absence of goiter and no prior neck dissection. The technique required three trocars; one 5 mm trocar inserted through the middle line of the neck for the 0-degree 5 mm endoscope, and two 3 mm trocars inserted laterally in order to perform a bilateral exploration. The neck was inflated to 10 mm Hg pressure with a low flow (3 L/min). RESULTS: Exploration was unilateral, bilateral and interrupted respectively in 52, 45 and 3% of the cases. The reasons for stopping were: an incidentally discovered thyroid carcinoma; moderate bleeding occurring from an anterior jugular vein after introduction of a lateral trocar; and a too-short neck. Parathyroid abnormalities were found in 86% of the patients (84 single adenomas, one double adenoma, one hyperplasia of the four glands). In 14% of the cases, the exploration was unsuccessful. A horizontal cervicotomy was required in 15% of the cases (14 negative explorations and one hyperplasia of the four glands). No intraoperative or postoperative complications occurred. The mean hospital stay was 24 hours. After 3-month follow-up, the serum calcium level was normal in 96% of the cases and cosmetic results were excellent. CONCLUSION: A video-endoscopic approach for parathyroidectomy is feasible and safe. With sufficient experience, a bilateral and complete exploration of the neck is possible. Therefore this technique represents a good alternative to the traditional cervicotomy in patients with uniglandular disease. PMID- 11413811 TI - [Ultrasonography of postoperatively recurrent varicose veins in the area of the short saphenous vein]. AB - STUDY AIM: The aim of this retrospective study was to classify postoperative recurrent varicose veins in the area of the short saphenous vein. PATIENTS AND METHOD: This retrospective ultrasound Doppler exploration was performed in 60 patients (77 limbs) who had been operated with crossectomy, isolated or associated with a stripping of the short saphenous vein, after a mean 9.2-year interval. RESULTS: Recurrences were classified in five categories: 14.8% of the patients had a recurrence in relation to a venous stump at the level of the crossectomy; 32.1% had a saphenous vein in its anatomical location, 21% had reflux due to incompetence; in 28.4%, recurrence was not correlated with the short saphenous vein; and in only 3.8%, there was a pseudo-angiomatosis appearance. In half of these patients, recurrence was related to an incomplete stripping of the short saphenous vein. CONCLUSION: In order to avoid incomplete and inefficient treatments leading to recurrent varicose veins, an ultrasound Doppler exploration is necessary before and after crossectomy and stripping of the short saphenous vein. PMID- 11413812 TI - [Experience with preperitoneal hernioplasty using Stoppa's procedures in the Ivory Coast]. AB - OBJECTIVE: The treatment of groin hernias with a high risk of recurrence often provides therapeutic difficulties in Africa for fear of using mesh due to the potential risk of sepsis. The aim of this retrospective study was to emphasize the simplicity and efficacy of Stoppa's technique. PATIENTS AND METHODS: From 1994 to 1999, 128 patients with 239 groin hernias underwent hernioplasty with Stoppa's technique. They were all men with a mean age of 54 +/- 8 years. They were classified in stages III and IV (46.8%, n = 60) of Nyhus. One hundred and eleven patients (86.7%) had bilateral hernias and 64 had a factor of recurrence (50%). RESULTS: Mean operating time was 150 +/- 42 min (50-240 min). Follow-up ranged from 6 months to 6 years, with a mean time of 26 months. Eighteen patients were lost to follow-up. The mortality rate was 0.8% (n = 1). The morbidity rate was 18.7% (n = 24) with five hematomas (3.9%), 11 seromas (8.6%), one parietal infection (0.8%) and one recurrence (0.8%). CONCLUSION: These results suggest that Stoppa's technique is efficient and technically feasible in our conditions and has to be performed more frequently. PMID- 11413813 TI - [Experimental study of monopolar electrical and ultrasonic dissection]. AB - The aim of this experimental study was to compare the disadvantages of a monopolar electric scalpel and an ultrasonically-activated scalpel in a porcine model. Using thermal gradients, the ultrasonically-activated scalpel significantly decreased thermal energy dispersion to surrounding structures and thus decreased the risk of injuries to adjacent tissues and nerves, as opposed to the monopolar electric scalpel. Histological examination of nerves showed that necrosis was significantly less important with the use of the ultrasonically activated scalpel. This experimental study demonstrated that the ultrasonically activated scalpel caused less thermal and histological damage than the monopolar electric scalpel. PMID- 11413814 TI - [Acute pancreatitis after bilateral laparoscopic adrenalectomy in patients with ectopic ACTH syndrome]. AB - Two cases of acute necrotizing pancreatitis after bilateral laparoscopic adrenalectomy were observed in patients with an ectopic ACTH syndrome. Two reasons may be suspected: the difficulty of dissection in such patients and the specific morbidity in relation to hypercorticism. PMID- 11413815 TI - [Blunt force rupture of the thoracic aorta and diaphragm: an unusual association]. AB - The aim of this study was to report two cases of rupture of the right hemidiaphragm with intra-thoracic liver hernia, associated with a traumatic aortic disruption. These two lesions followed traffic accidents, and were both treated separately. Both patients had a long-term favourable progression. Association of these two lesions is particularly rare: fewer than 50 cases have been previously described in an extensive review of the literature. CT-scan seems to be particularly efficient in diagnosis, even if less efficient than other more specific explorations. The order of surgical management is guided by the physical examination, and especially by abdominal emergency. PMID- 11413816 TI - [Surgery responsibilities and costs]. AB - This study on surgeons' medical-legal responsibility was based on a retrospective analysis of closed files involving surgeons insured by Le Sou Medical. A total of 122 files of bodily injury settled during 1998 were studied; 32% of the files in this series which had been settled in or out of court were found to be at fault; the total cost was over 7 million francs. The analysis of these files has shown that the majority of accidents resulting in legal proceedings could have been avoided. PMID- 11413817 TI - [The Hotel-Dieu de Lyon, the most beautiful hospital in the kingdom during the reign of Louis XV]. AB - The Hotel-Dieu was built between 1741 and 1761 on the banks of the Rhone, composed of a magnificent two-story facade, and the whole crowned by a dome which added to the glory of the city. For this grandiose and costly project, Soufflot was called in. The Hotel-Dieu became the best hospital in the realm. The healthcare personnel consisted of 130 hospitaller sisters and 50 brothers under the guidance of the chaplains. Eight to ten journeymen surgeons, forefathers of the interns, were spread throughout the wards. The principal surgeon chosen by the rectors became, after 6 years of practice, a fully qualified surgeon, or a 'surgeon gaining mastery', and could then practice freely in the city. In 1755, 1300 patients were hospitalized in the old and new buildings. PMID- 11413818 TI - Antifungal actinomycete metabolites discovered in a differential cell-based screening using a recombinant TOPO1 deletion mutant strain. AB - In the course of a natural product screening for inhibitors of fungal topoisomerase 1 (TOPO 1), extracts from the actinomycete strains WS 1410 and BS 1465 exhibited promising activities. Bioguided fractionation of the culture broth by preparative HPLC methods yielded the collismycins A (1) and B (2) as active principles of strain WS 1410. Out of the mycelial extracts of strain BS 1465 the bioactive new natural products, cyclo-homononactic acid (3) and cyclo-nonactic acid (5) and the structurally related but inactive homononactic acid (4), were isolated. Both collismycin isomers inhibited the recombinant yeast strains ScAL 141 and ScAL 143 (TOPO 1 deletion mutant) in a non-specific manner with an MIC in the range of 2 micrograms/ml. The novel cyclo-homononactic acid (3) and cyclo nonactic acid (5) showed higher selectivity towards the wild type strain (MIC = 2 micrograms/ml as compared to 10 micrograms/ml for the deletion mutant). All compounds obviously address a target other than TOPO 1 since they do not exhibit activities in a concurrent TOPO 1 enzyme assay. PMID- 11413819 TI - Synthesis and antimicrobial activity of some new 2-phenyl-N-substituted carboxamido-1H-benzimidazole derivatives. AB - Some 1H-benzimidazole-carboxamide derivatives were prepared and their antimicrobial activities against Staphyloccus aureus, Escherichia coli and Candida albicans evaluated. Compounds 18, 22, and 25 exhibited the best activity against Candida albicans. PMID- 11413820 TI - Phenylamides of 1-phenyl (or methyl)-5-benzamidopyrazole-4-carboxylic acid as vratizolin analogs with analgesic and antiinflammatory activities. AB - A number of phenylamides of 5-benzamidopyrazole-4-carboxylic acid were prepared in 50-80% yields from 1-phenyl (or methyl)-6-phenylpyrazolo[3,4-d]1,3-oxazin 4(1H)-ones and aniline derivatives. All the compounds were tested for their analgesic and antiinflammatory activities, as well as for their ulcerogenic potential and acute toxicity. Some derivatives, when compared to phenylbutazone, proved more active in the tests for analgesic and antiexudative activities, but less active in the carrageenin paw oedema test. The compounds proved to posses marginal or no ulcerogenic effect, as well as low systemic toxicity. PMID- 11413821 TI - Synthesis, physicochemical characterization, and cytotoxic screening of new zirconium complexes with coumarin derivatives. AB - Zirconium complexes of mendiaxon, warfarin, coumachlor, and niffcoumar have been synthesized by reaction of the ligands with zirconium chloride in stoichiometric ratio 1:2. The formation of the complexes has been proved on the basis of elemental analysis, IR-spectroscopy, 1H-NMR spectroscopy, and thermal studies. Differential thermal analyses and thermogravimetric analyses have been applied to study the compositions of the new complexes. It is concluded that the lactone- and the keto-carbonyl groups of warfarin, coumachlor, and niffcoumar are bonded to the metal ion as bidentate ligands, but mendiaxon is bonded as monodentate ligand. Cytotoxic screening by MTT-assay was carried out. Among these compounds the zirconium complex of mendiaxon showed highest cytotoxic activity against human promyelocytic leukemic HL-60 cells. The inorganic salt was found to be active against this cell line. PMID- 11413822 TI - Aryl rings are part of the darpone pharmacophore. AB - In order to investigate structure-activity relationships in the antiproliferative darpone series, the derivatives 8a and 8b lacking the aryl substituents in either 2- or 4-position were synthesized and evaluated for cancer cell growth inhibition. Since both 8a and 8b were devoid of noteworthy antiproliferative activity, both aryl substituents can be considered as part of the darpone pharmacophore. PMID- 11413823 TI - beta-lactam derivatives as enzyme inhibitors: carboxy peptidyl derivatives of (S) 4-oxoazetidine-2-carboxylate as peptidomimetics. AB - 4-Oxoazetidine-2-carboxylic acid, protected at the nitrogen by silyl groups, was coupled with amino acid and oligopeptide esters. Desilylation and deprotection of the amino acid residues yielded the free beta-lactam peptides. Structure and properties were elucidated by spectroscopic methods and discussed. Some selected compounds were tested as fibrinogen inhibitors and for thrombocyte aggregation. None of the compounds showed any activity up to a concentration of 10(-5) Mol/l. Some other compounds exhibited a weak inhibitory activity against elastase (PPE). PMID- 11413824 TI - Synthesis and biological activities of diquaternary dipiperazinium salts containing dithiocarboxyl groups. AB - A series of diquaternary dipiperazinium salts containing dithiocarboxyl groups 6a f and 9 were synthesized and evaluated for their analgesic and sedative activities. The result showed that the presence of two quaternary ammonium cations and the distance between them are very important for the activities of the salts. Compound 6b exhibited the best activities (at dose 2 mg/kg, analgesic, 57%; sedative, 59%) among compounds 6a-f. Compound 9 not only showed the most potent analgesic (85.4%, dose 1 mg/kg) and sedative (93.1%, dose 1 mg/kg) activities, but also exhibited anticancer activity against KB (68.7%, dose 10 microM). PMID- 11413825 TI - Synthesis of imidazo[4,5-d]oxazolo[3,4-a]pyridines. New heterocyclic analogues of lignans. AB - The preparation of new analogues of lignans carrying an imidazole ring has been achieved. Starting from L-histidinol, cis and trans stereoisomers have been obtained. The synthesized products lack the cytotoxicity displayed by related podophyllotoxins and azatoxin. PMID- 11413826 TI - Chemical and microbial characterization of household graywater. AB - In arid areas, the search for efficient methods to conserve water is of paramount importance. One of the methods of water conservation available today is graywater recycling--the reuse of water from the sinks, showers, washing machine, and dishwasher in a home. The purpose of this project was to characterize the chemical and microbial quality of graywater from a single-family home with two adults. Water samples from a graywater holding tank were analyzed over a seven month period for total coliforms, fecal coliforms, fecal streptococci, Staphylococcus aureus (S. aureus), Pseudomonas aeruginosa (P. aeruginosa), and coliphages. The pH, turbidity, biological oxygen demand (BOD), suspended solids (SS), electrical conductivity (EC), sulfates (SO4), and chlorides (Cl) were also measured. The mean numbers of total coliforms, fecal coliforms, fecal streptococci, and P. aeruginosa were 8.03 x 10(7), 5.63 x 10(5), 2.38 x 10(2), and 1.99 x 10(4) CFU/100 mL, respectively. S. aureus and coliphages were not detected. In the chemical analysis, mean values of 7.47 for pH, 43 nephelometric turbidity units (NTU) for turbidity, 64.85 mg/L for BOD, 35.09 mg/L for SS, 0.43 mS/cm for EC, 59.59 mg/L for SO4, and 20.54 mg/L for Cl were measured. These data were compared to data taken in 1986 and 1987, when two adults and one child lived in the household. Analysis showed no statistically significant difference in levels of total coliforms and suspended solids between the two data sets. There were statistically significant differences in levels of fecal coliforms, pH, turbidity, chlorides, sulfates, and BOD between the two households. Fecal coliforms, turbidity, and BOD were higher in the household with two adults and one child. Levels of Cl, SO4, and pH were higher in the household with two adults. PMID- 11413827 TI - Spray scrubbing of the nitrogen oxides into NaClO2 solution under acidic conditions. AB - The operating conditions of this study were closed to the typical operating conditions of flue gas desulfurization system in the coal-fired power plant. The objective of this study was to investigate the absorption performance of lean NO in an aqueous solution of acidic sodium chlorite using a bench-scale spraying column. The NO conversion and NOx removal efficiency were increasing with the increasing NO concentration, retention time, sodium chlorite concentration, operating temperature, and decreasing initial pH of solution. As the sodium chlorite concentration were higher than 0.4 M, the NO conversion and NOx removal efficiency were 100% and 80%, respectively. The NO conversion and NOx removal efficiency under initial pH 4-7 were higher than that of initial pH > 7. It meant that this process might be suitable to combining with traditional wet flue gas desulfurization system. As the NO2/NOx ratio in the effluent gas was closed to 0.5, it might be suitable to be absorbed in the second scrubbing column operated under alkaline condition. PMID- 11413828 TI - Composting of solid and sludge residues from agricultural and food industries. Bioindicators of monitoring and compost maturity. AB - A study to monitor the composting process, to evaluate the effectiveness of bioindicators for the quality and maturity of cured compost obtained by a mixture of winery residues, sludges from dairies and solid residues from food processing (grape-stalks, grape-dregs, rice husks), was conducted. Composting process lasting five months was monitored by chemico-physical, spectroscopic (FTIR, DTG and DSC), microbiological and enzymatic analyses. Biological activities (ATP, DHA contents and several enzymatic activities), impedance variations (DT) of mixed cultures during growth and potential pathogens (E. coli and Salmonella sp.), were determined. The phytotoxicity tests gave a germination index higher than 90% and no significant genotoxic differences between controls and the compost samples were evidenced. Pathogens were not found on the cured compost that can therefore be satisfactorily used as amendment for agricultural crops. However, no single measurement of a composting process factor, biological, chemical or physical, gave a comprehensive view of the quality of a specific composting. We proposed a tool of bioindicators of potential activity and markers in combination for integrated evaluation of monitoring of composting process and compost quality. The responses of several enzymatic activities were positive and indicative of their favorable use capable to reveal even very small changes within microbial population and activity in test and monitoring of compost programmes. PMID- 11413829 TI - A column study of soil contamination by lead: influence of pH and carbonate content. I. Experimental results. AB - The influence of soil carbonate content on the fate of lead in soil was studied in a lab-scale column under different pH values of the contaminant solution. Results indicated that retention of this toxic heavy metal (up to 38% weight at pH = 5) occurred which was proportional to the total carbonate content. A decrease in the pH of the aqueous solution entering the column resulted in a decrease of the retention of lead in the soil. Furthermore, the concentration of lead in the effluent was increased dramatically. Concentrations of lead about 2.5 times higher than in the contaminant solution were measured at pH = 3. Knowledge of these phenomena is important for risk assessment and remediation feasibility studies. PMID- 11413830 TI - Toxicity and biodegradation of products from polyester hydrolysis. AB - Toxicity of products from polyester hydrolysis such as succinic acid (SA), adipic acid (AA), mandelic acid (MA), terephthalic acid (TA), 1,4-butanediol (1,4-B), ethylene glycol (EG), styrene glycol (SG) and 1,4-cyclohexane dimethanol (1,4-C) was evaluated by phytotoxicity test on germination of young radish seeds and by cytotoxicity test on HeLa cells. The phytotoxicity test revealed SG > MA > 1,4-C > AA approximately SA > TA approximately EG > 1,4-B in order of decreasing toxicity taking into consideration the growth behavior after germination as well as the percentage of germination. Toxicity on HeLa cells decreased in slightly different order compared to that on young radish seeds, i.e. SG > 1,4-C > MA > TA > SA > AA > EG > 1,4-B. Tests for the phytotoxicity and for cytotoxicity indicated that the aromatic compounds were more harmful than the aliphatic ones. Each group of 4 strains which grew most rapidly on each agar plate containing SA, AA, MA, TA, 1,4-B, EG, SG and 1,4-C respectively as a sole carbon source was identified by the fatty acid methyl esters analysis. The modified Sturm test was carried out using the single isolated strain, an activated sludge or a mixed soil to measure the rate of mineralization of the compounds into carbon dioxide. The aliphatic compounds were mineralized more easily than the aromatic compounds. 1,4 C showed the most exceptionally slow degradation. A scrutiny of residual 1,4-C after degradation is required before polyesters containing 1,4-C could be classified into compostable because 1,4-C has detrimental effects on young radish seeds and HeLa cells and has a tendency to accumulate in the environment due to its slow degradability. PMID- 11413831 TI - Polonium 210Po in cigarettes produced in Poland. AB - This paper discusses the results of 210Po determinations in the fourteen most frequently smoked brands of cigarettes, which constitute over 80% of total cigarette consumption in Poland. The 210Po activity in the cigarette samples analysed (tobacco, ash, filter before and after smoking) were measured using alpha spectrometry (Canberra-Packard). The data indicates that there is considerable variation in the polonium content of these brands. The highest 210Po content per sample was found in the cheap "Popularne" brand (24.12 mBq), the lowest in "Caro" (4.23 mBq). There was also a large difference between the polonium remaining in the ash in comparison with its total content in the tobacco in all the brands (from 4.3% for "Golden American" to 71.0% for "Sobieski King Size"). The analysis has demonstrated that filters absorbed only a small amount of the polonium contained in the tobacco. "Caro" cigarettes have the most efficient filter, retaining 25.1% of the polonium contained in the tobacco, but most filters absorbed only 0.1-7.2% of polonium. The daily inhalation of 210Po by Polish smokers who get through one pack per day ranges from 20 to 215 mBq, but people smoking two or more packs of "Popularne" brand will inhale over 430 mBq of polonium per day. PMID- 11413832 TI - Formation and evolution of haloacetic acids in drinking water of Beijing City. AB - The formation and evolution of haloacetic acids (HAAs) in drinking water were examined in five water treatment plants in Beijing City, China. It was found that HAAs in water increased due to prechlorination and postchlorination. The conventional treatment processes removed the HAAs in less than 20%. By comparison, activated carbon bed removed 49-86% of HAAs and was considered to be an effective treatment unit for HAAs removal. The HAAs concentration in distribution system was associated with water sources, especially the total organic carbon in raw water, while its fluctuation appeared small. PMID- 11413833 TI - Disinfection by-products and volatile organic compounds in the water supply system in Athens, Greece. AB - On a monthly basis, on a one year period, disinfection by-products (DBPs) and volatile organic compounds (VOCs) were measured in raw water and in treated water of four water treatment plants (WTP) as well as from eight representative points of the distribution network of Athens. Liquid-liquid extraction (LLE) method followed by gas chromatography (GC) with electron capture detection (ECD) and mass spectrometry (MS) was used for the determination of the DBPs, while the purge-and-trap (PAT) technique followed by GC-MS was used for the determination of the VOCs. The DBPs that have been studied included trihalomethanes (THMs), haloacetonitriles (HANs), haloketones (HKs), chlorate hydrate (CH) and chloropicrin (CP), while forty-one VOCs were also studied including THMs. Chloroform (CM), dichlorobromomethane (DCBM), chlorodibromomethane (CDBM) were the major organic compounds found in all treated water samples. HANs, HKs, CH and CP were detected in treated samples but at lower concentrations. PMID- 11413834 TI - The accumulation effect of lead on DNA damage in mice blood cells of three generations and the protection of selenium. AB - The accumulation effect of lead on DNA damage and the protection of selenium against lead were studied in mice blood cells of three generations. The test animals were divided into four groups: controls, Pb group, Se group, Pb + Se group. Lead and Selenium were administered to mice in the drinking water. The concentrations of lead (in form of lead acetate) and selenium (in form of Kappa selenocarrageenan) used in this paper were 1 microgram/ml and 0.25 microgram/ml respectively. The percentage of damaged cells (DNA comets) was examined and detected conveniently by the single-cell gel electrophoresis (comet) assay. Results showed that 1 microgram/ml lead did not damage the female mice blood cells of the first generation seriously. However, there was a significant damage effect in both sexes of the second and third generations, suggesting the accumulation effect of lead was very significant starting from the second generation. In addition, under normal physiological conditions, Kappa-Se did not appear to enhance the protective ability on DNA damage significantly, whereas under lead exposure conditions, administration of Kappa-Se displayed the occurrence of a good protection against lead intoxication which was started from the female mice of the second generation. PMID- 11413835 TI - Feasibility of fluidized-bed bioreactor for remediating waste gas containing H2S or NH3. AB - Pseudomonas putida for H2S and Arthrobacter oxydans for NH3 were immobilized with Ca-alginate and packed inside glass columns to form fluidized-bed bioreactors. The feasibility of the lab-scale bioreactor for the treatment of H2S or NH3 was examined. Phosphate salt, being added to the nutrient solution as buffer solution, may chelate with Ca2+ in the Ca-alginate beads, resulting in the disintegration of gel structure. When the buffer capacity of the phosphate solution was over the critical point of 33.5 mM/pH, all calcium ions in the bead were released and beads were broken. Increasing liquid flowrate and inlet gas concentration favored to H2S and NH3 removal. Carbon source addition was essential and facilitated malodorous removal for this system. Removal capacity increased with inlet concentration. However, increasing pattern was dependent of H2S or NH3. The result clearly indicated that bioreactor was suitable to be applied for the industry of livestock farm for removing wastegas containing H2S or NH3. PMID- 11413836 TI - Estimation of atmospheric mixing heights using data from airport meteorological stations. AB - In this study, ground and sounding meteorological data at the Beijing Airport during 1991 to 1995 were used for estimating the local atmospheric mixing heights. Three methods were compared for this purpose, including the dry adiabatic method, the Nozaki model, and a modified Nozaki model. The modification of the Nozaki model included joint frequencies for wind-velocity and stability based on the complexities of local meteorological conditions. The estimated values from the three methods were verified through the data measured by the Beijing Meteorological Center. The results indicated that the dry adiabatic method has the best performance. The modified Nozaki model was better than the commonly used. This study is a new attempt in utilizing airport meteorological data to estimate atmospheric mixing heights. PMID- 11413837 TI - Effect of relative humidity on mixed aerosols in atmosphere. AB - In this study, the effects of relative humidity on the deliquescent point and size of internally mixed aerosols diameter, NH4NO3 and (NH4)2SO4 were investigated using a Tandem Differential Mobility Analyzer (TDMA) with a relative humidity conditioner. The growth of mixed aerosols appears to have two deliquescent steps. The first one was at about 61.2-61.3%, but the second one was at around 77-78%. At the first deliquescence point, growth ratio at phase change was 7.5%, which agrees with the growth ratio of ammonium nitrate aerosol. Growth ratio of phase change at the second deliquescence point was about 20%, lower than the growth ratio of ammonium sulfate aerosol. In the relative humidity range of 80-85%, the growth ratio of the mixed aerosols reached 60%. In other words, it appears that growth ratio increases with the size of aerosol. Furthermore, a theoretical growth model of mixed aerosols was developed and applied to estimate the amount of composition of the mixed aerosols dissolved at each deliquescence point. The results also show that some of ammonium sulfate already dissolved at the first deliquescence point according to the theoretical growth model. PMID- 11413838 TI - Application of various bod test methods in landfill leachates. AB - This study examined the determination of BOD in landfill leachates by dilution (D Method) and manometric methods (M-Method). The discrepancies between these methods were discussed and statistical tests were carried out. The effects of sample dilution and seeding on BOD results were examined. In comparison to the D Method the M-Method was found to be more sensitive to increases in chloride and Total Kjeldahl Nitrogen (TKN) concentrations in leachate samples. However, in the M-Method the positive interference of nitrogenous BOD (NBOD) to carbonaceous BOD (CBOD) was more successfully prevented. The BOD rate constant k and the ultimate BOD (BODu) were estimated by non-linear regression. With the M-Method these parameters could be more reliably estimated than the D-Method. Based on these results, suggestions were made for BOD analyses in landfill leachates in future studies. PMID- 11413839 TI - The use of Mg/Al layered double hydroxide for color removal of textile wastewater. AB - Layered double hydroxide of Mg-Al-carbonate system (MACH) was prepared and its heat-treated product (MACHT) was obtained by calcination at 500 degrees C. The resulting materials were used as an adsorbent for removal of color from synthetic textile wastewater (STW) and textile wastewater (TWW). Batch kinetic study showed that these materials are an efficient adsorbent for textile dye. The maximum adsorption capacities between 16 to 32 mg of dyes per g of adsorbent was obtained by fitting the adsorption data to the Langmuir adsorption Isotherm. It was found that the adsorption capacity of MACHT is higher than MACH. PMID- 11413840 TI - Effects of hydraulic backwash load on effluent quality of upflow BAF. AB - Using an upflow biological aerated filter (BAF) with treatment capacity of 100 ton/day, various hydraulic loads of backwashing water and influent were investigated to determine how they affect effluent quality of BAF during normal operation. The BAF packed with expanded polypropylene media of 3-5 mm diameter was operated over a five-month period for paper wastewater treatment. The average removal efficiencies of 78% for chemical oxygen demand (COD) and 88% for suspended solids (SS) were maintained, but the effluent CODs varied between 5-60 mg/l depending on the biodegradable fraction of influent wastewater. During normal BAF operation with cyclic backwash, the effluent SS concentrations showed initial peaks after backwash and gradually decreased to a steady state, while the soluble CODs of effluent did not significantly changed. The effluent SS concentrations and the required time intervals to reach steady state after backwash were related with hydraulic loads of backwashing water and influent wastewater applied to the BAF. A higher load of backwashing water gave larger deviation of initial SS peaks from the steady state, the extent of which was more significant under the higher load of influent wastewater. PMID- 11413841 TI - Step feeding strategy for enhancing nitrogen and phosphorus removal in AOAO process. AB - This study describes a biological process containing anaerobic/oxide/anoxic/oxide (AOAO) stages, which incorporates a stepwise feeding strategy to enhance nitrogen and phosphorus removal efficiencies. In the experimental reactors, synthetic wastewater was fed into the anaerobic stage and anoxic stage separately, while the substrates and nutrients were successfully consumed without recycling either nitrified effluent or external carbon sources. Experimental results demonstrated that the anoxic stage was provided an intrinsic sufficient carbon source and caused NOx (NO2-N + NO3-N) concentration to reduce from 11.85 mg/l to 5.65 mg/l. The total Kjeldahl nitrogen (TKN) removal rate was between 81.81% and 93.96%, while the PO4-P removal ratio was between 93% and 100%. The substrate fed into the anaerobic at flow rate Q1 and into the anoxic at flow rate Q2. In addition, three different ratios were performed with Q1/Q2 varied from 7/3, 8/2 to 9/1. The AOAO process clearly saves nearly one-third of the energy compared to the typical Biological nutrient removal (BNR) system A2O process. PMID- 11413842 TI - [Otorhinolaryngologic pain in the adult. A comparison of the analgesic efficacy of lysine acetylsalicylate and paracetamol]. AB - OBJECTIVE: Unlike paracetamol, aspinin has anti-inflammatory properties which may be helpful in relieving pain associated with inflammation. To confirm this hypothesis, we performed a randomized, double-blind, double-dummy study to compare the relative analgesic efficacy of lysine acetylsalicylate (LAS) and paracetamol (PAR) in ENT pain in adults. PATIENTS AND METHODS: After initiation of the treatment by a placebo (PLA), both drugs were given at the same dose (1 g twice daily on D1 and 1 g three times daily on D2) for two days. From the third day to the seventh day, the patients could freely take the same drug if necessary. The analgesic effect was assessed by measuring the area under the curve (AUC) of pain severity as scored by patients on a visual analogue scale. The secondary efficacy criteria were the difference relative to the basal pain score of the scores at each evaluation time (PID), the sum of these differences (SPID), and the proportion of patients responding to the treatment (i.e. experiencing a decrease of at least 50% in the pain score). RESULTS: A total of 312 patients, included by 30 French centres, were treated by LAS (n = 156) or PAR (n = 156). Pain was related to an infection in 98% of the cases. Nine percent of patients in each treatment group were responders to the PLA and were consequently not included in the per protocol (PP) analysis but in the intent-to-treat (ITT) analysis. During days 1 and 2, patients in the LAS group had less pain than those of the PAR group, the difference becoming significant at Day 2 whatever the population analysed, PP (p = 0.01) or ITT (p = 0.01). LAS also showed better analgesic efficacy than PAR as assessed by measures of PID and SPID, the difference being significant at several times of evaluation for PID, and on Day 2 for SPID evaluation in the PP (p = 0.01) and ITT analysis (p = 0.007). The proportion of responders was significantly higher in the LAS group than in the PAR group (70% versus 56%, p = 0.01). Safety was comparable between the treatment groups. The overall efficacy judgement of investigators and patients was significantly better for LAS than for PAR (p = 0.01 and p = 0.006 respectively, at the Day 2 evaluation). CONCLUSION: In this study, LAS proved to be more effective than PAR in the treatment of ENT-associated pain in adults. This is probably related to the anti-inflammatory activity of LAS. PMID- 11413843 TI - [Anticancer chemotherapy, risk factor for infection at the operated site in cervicofacial surgery]. AB - BACKGROUND: In order to evaluate occurrence and risk factors for wound infection (WI) in head and neck uncontaminated surgery, we carried out a prospective study. METHODS: From january 1997 through january 1999, we prospectively evaluated 212 wounds of all patients having uncontaminated head and neck surgery at the Oscar Lambret Cancer Center (neck dissections, parotidectomies, thyroidectomies, explorative cervicotomies, cutaneous resections). No antibiotic prophylaxis was given. WI was defined as a wound with pus. Statistical evaluation was performed using the chi 2 test. In univariate analysis, differences were considered significant p < 0.05. RESULTS: The overall WI rate was 6.6% (14/212). In univariate analysis, previous chemotherapy is the only risk factor for WI were: (p < 0.00001). Multivariate analysis was not performed. CONCLUSION: Like other cancer locations, chemotherapy was a major risk factor for WI. In these cases, a phase ill trial could confirm efficacy of standard antibiotic. PMID- 11413844 TI - [Infection of a pneumonectomy site 16 years after intervention]. AB - BACKGROUND: Infection of the residual cavity after pneumonectomy generally occurs early after surgery. CASE REPORT: A 67-year old patient was hospitalized with fever 16 years after pneumonectomy for lung cancer. Investigations led to the diagnosis of infection of the pneumonectomy cavity. DISCUSSION: Late infection several years after pneumonectomy is exceptional and usually occurs in a setting of bacteriemia. Diagnosis is difficult due to modifications of the thoracic signs but should be entertained whenever unexplained fever or an inflammatory syndrome occurs in a pneumonectomized patient. The thoracic CT scan shows an abnormally enlarged cavity. Bacteriological examination of evacuated fluid provides the key to diagnosis. Drainage-lavage is indicated. Surgery may be needed exceptionally. PMID- 11413845 TI - [Horton disease with with right orbital edema]. PMID- 11413846 TI - [Lithium poisoning: severe neurologic complications]. PMID- 11413847 TI - [Secondary hypertrophic osteoarthropathy in tumor of the pancreas]]. PMID- 11413848 TI - [Bordetella bronchiseptica infection in a Senegalese HIV positive patient: carrier state or illness?]. PMID- 11413849 TI - [Exercise which saves or exercise which kills?]. PMID- 11413850 TI - [Celiac disease, magnesium deficiency and venous thrombosis]. PMID- 11413851 TI - [Has hepatitis C become curable by a new pegylated interferon?]. PMID- 11413852 TI - [Intra-familial graft. Respecting public health regulation and professional confidentiality may be incompatible]. AB - Under specific conditions, French law authorizes organ donation despite the donor's seropositivity for certain infectious conditions. The recipient must however be informed of the potential risk of graft-related infection. Anonymous donation being the rule, the rights of the donor are respected since his/her serological status remains a medical secret. However, an exception to the rule of anonymous donation is allowed for organ donation between family members. In such a situation--often justified by the urgent nature of the transplantation--the donor's serological status would have to be revealed to the recipient, breaking the rule of medical secrecy. The physician who breaks the rule is simply implementing legal regulations (with the subsequent protection against any penal or disciplinary measures) but nevertheless performs an ethically questionable act. The recompense for donation would be an incongruent violation of personal rights. At the present time, there does not appear to be a satisfactory solution to this dilemma. The only solution that could be put forward would be to ask the donor to voluntarily inform the recipient of his/her seropositivity. PMID- 11413853 TI - [Blood lipid abnormalities during treatment with protease inhibitors]. AB - HYPERLIPIDEMIA: HIV-1 protease-inhibitors therapy is associated with increased levels of triglycerides, LDL-cholesterol and Lp(a). But the understanding of hyperlipidaemia occurring in patients treated with HIV-1 protease-inhibitors is not easy since HIV infection itself is associated with lipid abnormalities and since HIV-1 protease-inhibitors therapy is also responsible for the development of a lipodystrophy syndrome (insulin resistance) which may influence lipid metabolism. However, many data indicate that HIV-1 protease-inhibitors therapy itself modifies significantly lipid metabolism. UNDERLYING MECHANISMS: The mechanisms involved in HIV-1 protease-inhibitors induced hyperlipidaemia are still unclear. HIV-1 protease-inhibitors could bind to LRP (Low density lipoprotein receptor Related Protein), impairing hepatic chylomicron-remnants and VLDL uptake. They could interact with the retinoid X receptor (RXR), which functions as a heterodimer with PPAR (Peroxisome Proliferator Activator Receptor). HIV-1 protease-inhibitors could also modify lipoprotein metabolism through cytokines. PMID- 11413854 TI - [Reversible posterior leukoencephalopathy. A new syndrome or a new name for an old syndrome?]. AB - DEFINITION: Reversible posterior leucoencephalopathy syndrome is a recently proposed cliniconeuroradiologic category characterized by seizures, disorders of consciousness, visual abnormalities and headaches associated with predominantly posterior white matter abnormalities on CT and MRI examinations. MECHANISMS AND CAUSES: Since the initial description, reversible posterior leucoencephalopathy has been recognized in an increasing number of medical conditions, including hypertensive encephalopathy, eclampsia, and immunosuppressive treatment. The rapid resolution of clinical and neuroradiologic abnormalities suggests cerebral edema which is thought to result from impaired cerebrovascular autoregulation and endothelial injury. PROGNOSIS: Whereas reversibility of the posterior leucoencephalopathy is a characteristic of the disorder, this reversibility is contingent on prompt controlling blood pressure and discontinuing the offending immunosuppressive drug. PMID- 11413855 TI - [Chronic pancreatitis: diagnostic and therapeutic management]. AB - NATURAL HISTORY: Alcoholism is a common cause of chronic calcifying pancreatitis. Abdominal pain, weight loss and acute complications are the early manifestations. Besides chronic pain, about 80% of the patients develop acute pancreatitis, pseudocysts, compression of the main bile duct, serous effusion or digestive bleeding during the first five years of the disease course. Painful episodes are more frequent after 5 to 10 years, with a risk of jaundice and persistent pseudocyst formation. CHRONIC COURSE: Beyond 10 years, calcified stones and fibrosis progressively replace the pancreas parenchyma. Painful events become more parse and after 15 years, diabetes and exocrine insufficiency predominate. TREATMENT: Abstention from alcoholic beverages, analgesics administration and balanced diet are essential therapeutic elements. Insulin is required for the diabetes and pancreas extracts in gastroprotected microcapsules are needed to compensate for the chronic exocrine insufficiency. This medical treatment may be able to control pain but interventional radiology (pseudocyst drainage, antalgesic infiltration of the celiac plexus), interventional endoscopy (pseudocyst drainage, biliary stent, pancreatic endoprosthesis), or surgery (deviation of the bile ducts, wirsungojejunal derivation, exceptionally resection) may also be required. PMID- 11413856 TI - [Association of ampulloma and pancreas divisum, ultrasound endoscopic aspects]. PMID- 11413857 TI - [Intraesophageal foreign bodies, an emergency accessible to endoscopic treatment]. PMID- 11413858 TI - Nonadrenergic, noncholinergic responses of the human colon smooth muscle and the role of K+ channels in these responses. AB - The aim of the present study was to investigate the possible role of nitric oxide (NO) as a nonadrenergic, noncholinergic (NANC) mediator in human colon smooth muscle in vitro and to examine its possible interactions with K+ channels. In the presence of atropine (10(-6) M) and guanethidine (10(-5) M), electrical field stimulation (EFS, 1-10 Hz, 0.3 msec, 50 V) for 10 sec induced relaxations which were inhibited by tetrodotoxin (10(-6) M). In the presence of NG-nitro-L-arginine methyl ester (L-NAME, 10(-4) M), relaxations induced by EFS at 1, 2, 4, 8 and 10 Hz were reduced by 38.7 +/- 4.3, 31.5 +/- 3.8, 54.3 +/- 5.4, 59.8 +/- 4.5 and 68.6 +/- 5.3%, respectively. The relaxations inhibited by L-NAME were restored by the preincubation of L-arginine (L-ARG, 10(-3) M) at all frequencies tested. D Arginine (D-ARG, 10(-3) M) had no effect. Tetraethylammonium (TEA, 10(-4) M) or glibenclamide (10(-6) M) significantly decreased the relaxations induced by EFS. Exogenously applied sodium nitroprusside caused concentration-dependent relaxation with maximum relaxation observed with 10(-3) M. TEA (10(-4) M) and glibenclamide (10(-6) M) significantly depressed the maximum response to sodium nitroprusside. In conclusion, our data indicate that NO is involved in NANC nerve mediated relaxation in the human colon smooth muscle and the relaxant responses to endogenously released or exogenously applied NO are mediated, in part, by activation of calcium-dependent and ATP-sensitive K+ channels. PMID- 11413859 TI - Alpha 2-adrenoceptor activity induces the antidepressant-like glycolipid in mouse forced swimming. AB - Antidepressants show an antidepressive-like behavioral effect on mouse forced swimming via induction of GalNAc alpha 1-3GalNAc-lipid. In order to detect the mechanisms of the neuron receptor activities on the effect, the glycolipid reactivities in the serum induced by tandospirone, a specific 5-HT1A receptor agonist, and clonidine, a specific alpha 2-adrenoceptor agonist, were investigated. Both of these agonists showed dose-dependent behavioral effects which were decreased by pretreatment with yohimbine, a specific alpha 2 adrenoceptor antagonist. The glycolipid reactivities corresponded with the effects. These present findings strongly suggest that alpha 2-adrenoceptor activity in the brain mediates the induction of the antidepressant glycolipid in the forced swimming test. PMID- 11413860 TI - A characterization of myocardial infarction induced by thrombotic occlusion and a comparison with mechanical ligation of the rat coronary artery. AB - In the present study we examined two different methods for inducing myocardial infarction in rats. We previously developed an animal model of coronary artery thrombotic occlusion induced by a photochemical reaction, which occurs when rose bengal (a photosensitizer dye) is injected into the animal and is irradiated with green light. Arterial occlusion is thereby achieved nonmechanically. Using this method, we investigated the effect of thrombolytic intervention on myocardial infarct size. Infarct size was determined 24 h after the induction of myocardial infarction. When tisokinase (3 mg/kg), a native tissue-type plasminogen activator, was administered 3 min after the ST-segment elevation on a lead II electrocardiogram, the infarct size was 20.6 +/- 5.1%, which was significantly smaller than that of control rats (37.3 +/- 4.6%). When tisokinase was administered 10 min after the ST-segment elevation, the infarct size was 27.1 +/- 2.1%, which was not significantly smaller than that of controls. The rat coronary artery thrombosis model incorporates many aspects of coronary thrombosis. It differs from the coronary ligation model in that it lends itself to the study of thrombolytic agents on animal models of myocardial infarction. PMID- 11413861 TI - Estradiol inhibits astrocytic GFAP expression in an animal model of neuroinflammation. AB - There are many animal models for studying different aspects of neurodegeneration. Lipopolysaccharide (LPS) injected in rats intracerebroventricularly induces neuroinflammation quite similar to the inflammatory component of chronic neurodegenerative conditions such as Alzheimer's disease. We used this model to examine the effect of estradiol on neuroinflammation. LPS or pyrogen-free saline were injected intracerebroventricularly (i.c.v.) into the lateral ventricle of male Wistar rats and estradiol was administered (200 micrograms/kg s.c.) 48 h before or 24 h after LPS injection. LPS-induced body weight loss was partially postponed by the treatment, especially in the rats pretreated with estradiol. When analyzing GFAP glial cell morphology in the CA3c area of the hippocampus and corpus callosum, as well as the number of astroglial cells in CA3c and CA1, GFAP expression was found to be reduced. This was true especially in the animals pretreated with estradiol and to a much lesser extent in the posttreated ones. The data support the possible existence of a neuroimmunomodulatory effect of estradiol administration in neurodegenerative conditions, which influences the inflammatory component. PMID- 11413862 TI - Effect of omeprazole on the pharmacokinetics of sustained-release carbamazepine in healthy male volunteers. AB - The present study shows the effect of omeprazole administration on the pharmacokinetics of a sustained-release preparation of carbamazepine in healthy male volunteers. Multiple dose administration of omeprazole increased the Cmax, AUC0-infinity and elimination half life (t1/2e) of carbamazepine. The results suggest that drug monitoring should be carried out when carbamazepine is coadministered with omeprazole. PMID- 11413863 TI - Bioavailability of a crushed pantoprazole tablet after buffering with sodium hydrogencarbonate or magaldrate relative to the intact enteric coated pantoprazole tablet. AB - The aim of this study was to determine the bioavailability of orally administered, crushed pantoprazole tablets after buffering with either 1.4% sodium hydrogencarbonate or 1600 mg magaldrate relative to the intact enteric coated pantoprazole tablet. A single dose, three-period crossover study was performed with 18 healthy male volunteers. The crushed pantoprazole suspension, together with either sodium hydrogencarbonate or magaldrate, was administered via a nasogastral tube. Tmax of crushed pantoprazole was earlier as compared to the intact tablet (0.5 h vs. 3 h) and Cmax was approximately 10% higher due to faster absorption. The bioavailability of the crushed pantoprazole tablet relative to the intact pantoprazole tablet was 93% when using sodium hydrogencarbonate as the buffering agent, and 88% when using magaldrate. Based on the internationally approved confidence intervals for AUC (0.80-1.20) and Cmax (0.70-1.43), the crushed tablet buffered with sodium hydrogencarbonate was shown to be bioequivalent with the intact tablet (point estimates for AUC or Cmax: 0.93 or 1.10). For the crushed tablet buffered with magaldrate (point estimates for AUC or Cmax: 0.88 or 1.12) bioequivalence with the intact tablet could not be formally demonstrated, although the lower confidence limit for AUC of 0.78 was only slightly below the lower limit of the equivalence range of 0.80. PMID- 11413864 TI - Antioxidant status of the hypertrophic heart of Dahl hypertensive rat as a model for evaluation of antioxidants. AB - Development of hypertension, myocardial hypertrophy and the cardiac antioxidant status of male Dahl salt-sensitive (DSS) genetically hypertensive rats was evaluated and compared to that of normotensive Dahl salt-resistant (DSR) controls. In order to obtain exaggerated and more severe hypertension, half of the animals (10 per group) were Na loaded (8% NaCl diet) for 6 weeks. The myocardial antioxidant status was estimated in tissue homogenates on the basis of tissue glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). The results showed that 6 weeks of hypertension resulted in left ventricle myocardial hypertrophy, documented by weight, morphometry and morphological changes. The compromised myocardial antioxidant status of the DSS rats was defined by significantly decreased GSH-Px and glutathione activity (13% and 41%, respectively) as compared to DSR rats. SOD in DSS myocardium was increased by 47% compared to that in DSR myocardium, an effect that is considered a compensatory mechanism to the oxidative stress. All of the above changes were exaggerated by NaCl loading. It was concluded that DSS rats, on either a normal or high NaCl diet, displayed decreased antioxidant capacity, which is most likely genetically determined. Before the Dahl rat can be considered as a suitable model for testing new cardiac antioxidants, a full characterization of the level of cardiac oxygen free radicals is required. PMID- 11413865 TI - Adolescent sexual risk behavior: a multi-system perspective. AB - Adolescents are at high risk for a number of negative health consequences associated with early and unsafe sexual activity, including infection with human immunodeficiency virus, other sexually transmitted diseases, and unintended pregnancy. As a result, researchers have attempted to identify those factors that influence adolescent sexual risk behavior so that meaningful prevention and intervention programs may be developed. We propose that research efforts so far have been hampered by the adoption of models and perspectives that are narrow and do not adequately capture the complexity associated with the adolescent sexual experience. In this article, we review the recent literature (i.e., 1990-1999) pertaining to the correlates of adolescent sexual risk-taking, and organize the findings into a multisystemic perspective. Factors from the self, family, and extrafamilial systems of influence are discussed. We also consider several methodological problems that limit the literature's current scope, and consider implications of the adoption of a multisystemic framework for future research endeavors. We conclude with a discussion of the implications of the available research for practitioners working to reduce sexual risk behavior among adolescents. PMID- 11413866 TI - The roles of ethnicity and culture in the development of eating disturbance and body dissatisfaction: a meta-analytic review. AB - This meta-analysis involved 35 studies examining eating disturbance and body dissatisfaction in white and non-white populations and the role of acculturation in the development of eating-related psychopathology. While the role of acculturation in predisposing non-whites to eating disorders remains to be determined, mean effect sizes indicate that whites report more eating disturbance than non-whites. Differences are greatest when studies compare black and white college samples on measures of subclinical eating pathology, like dietary restraint, ideal body shape, and body dissatisfaction. They are weakest when non clinic populations and clinical forms of eating disturbance, like bulimia nervosa, are examined. These findings suggest that the current literature may be incorrect in its view that subclinical and clinical forms of eating disturbance represent the poles of a single continuum. In addition, they call into question the belief that SES influences the development of eating pathology. PMID- 11413867 TI - Interpersonal functioning among women reporting a history of childhood sexual abuse: empirical findings and methodological issues. AB - A subset of research exploring the long-term impact of child sexual abuse (CSA) has examined the adult interpersonal functioning of female survivors. The present review discusses empirical findings and critical methodological issues related to this important but often overlooked aspect of adult adjustment. Though characterized by several methodological limitations, this literature, as a whole, suggests that early sexual abuse represents a risk factor for a range of interpersonal dysfunction among female survivors, including problems with intimate partner relations, disturbed sexual functioning, and difficulties in the parental role. Suggested methodological improvements for future research include new approaches to the measurement of CSA and interpersonal variables, the need for comprehensive assessment of significant third variables, and the use of more representative sampling strategies. PMID- 11413868 TI - An examination of the diagnostic validity of dissociative identity disorder. AB - We review the empirical evidence for the validity of the Dissociative Identity Disorder (DID) diagnosis, the vast majority of which has come from research conducted within the last 10 years. After reviewing three different guidelines to establish diagnostic validity, we conclude that considerable converging evidence supports the inclusion of DID in the current Diagnostic and Statistical Manual for Mental Disorders. For instance, DID appears to meet all of the guidelines for inclusion and none of the exclusion guidelines; proposed by Blashfield et al. [Comprehensive Psychiatry 31 (1990) 15-19], and it is one of the few disorders currently supported by taxometric research. However, we also discuss possible problems with the current diagnostic criteria and offer recommendations, based on recent research, for possible revisions to these criteria. PMID- 11413869 TI - Psychophysiological arousal in older adults: a critical review. AB - In spite of the relatively high prevalence rates of anxiety disorders (AD) and related symptoms, very little is known about the experience, presentation, and assessment of anxiety in later life. Because the physiology of the autonomic nervous system changes with age, an enhanced understanding of how these developmental changes affect the somatic-physiological response patterns to anxiety-evoking stimuli among older adults may help explain whether we can generalize current assessment and treatment practices and procedures for AD from younger to older adults. In this paper, we describe and critically evaluate studies that have employed psychophysiological recording of autonomic arousal to anxiety-arousing or stressful stimuli among samples of younger and older adults. The conclusions one can draw from the review are quite limited by both the paucity of relevant literature and the methodological limitations of the published studies. PMID- 11413870 TI - Clinical and psychological features distinguishing obsessive-compulsive and chronic tic disorders. AB - Medical and biogenetic research has suggested that obsessive-compulsive disorder and chronic (multiple) tic disorder may share a common etiology. This article reviews corresponding evidence for psychological similarities and differences between the two disorders. There are similarities in self-management strategies, psychological traits (both report high scores on different aspects of perfectionism) and in the ego-syntonic-ego-dystonic cycle of the impulsive compulsive behavior. Situational cues likely to elicit or worsen the problem differ between the disorders as do associated emotions, comorbidity and background styles of action. In both disorders, cognitive factors, such as anticipations and appraisals of the problem, can play a role in onset and maintenance of the problem, and this raises the question as to whether cognitive or behavioral factors are best addressed in treatment. Psychological characteristics, such as lack of confidence, may contribute to apparent performance deficit. Psychological evaluation, particularly functional analysis, may aid in differential diagnosis between the two disorders, lead to improvement in treatment matching, and in understanding of the multidetermined etiology. PMID- 11413871 TI - Media violence and the American public. Scientific facts versus media misinformation. AB - Fifty years of news coverage on the link between media violence and aggression have left the U.S. public confused. Typical news articles pit researchers and child advocates against entertainment industry representatives, frequently giving equal weight to the arguments of both sides. A comparison of news reports and scientific knowledge about media effects reveals a disturbing discontinuity: Over the past 50 years, the average news report has changed from claims of a weak link to a moderate link and then back to a weak link between media violence and aggression. However, since 1975, the scientific confidence and statistical magnitude of this link have been clearly positive and have consistently increased over time. Reasons for this discontinuity between news reports and the actual state of scientific knowledge include the vested interests of the news, a misapplied fairness doctrine in news reporting, and the failure of the research community to effectively argue the scientific case. PMID- 11413872 TI - The family dynamics of intellectual development. AB - Birth order effects on intellectual performance show both positive and negative results. The confluence model reconciles these conflicting data by proving that these effects interact with the age of participants at testing, such that young children should show negative or no effects, whereas older individuals (past age 11 +/- 2 years) should show positive effects. Birth order studies strongly support this prediction. Some writers have claimed the apparent relation between birth order and intelligence is an artifact created by applying a cross-sectional analysis to data that should have been analyzed by comparing siblings within families. However, if siblings within the same family are compared all at the same time, their ages are necessarily different. As a result, birth order effects are confounded with age effects. Moreover, within-family data conceal patterns of aggregate effects that cross-sectional data reveal. PMID- 11413873 TI - Number of siblings and intellectual development. The resource dilution explanation. AB - The resource dilution model posits that parental resources are finite and that as the number of children in the family increases, the resources accrued by any one child necessarily decline. Siblings are competitors for parents' time, energy, and financial resources and so the fewer the better. Even one sibling is too many. The author describes the general elements of the dilution position and assesses its merits for explaining the effect of siblings on one component of the educational process--tests of intellectual development. The author identifies critical flaws in recent critiques of the dilution position and concludes that dilution continues to provide the most promising explanation for why children with few siblings score higher on tests of cognitive skills than children with many siblings. PMID- 11413874 TI - What causes birth order-intelligence patterns? The admixture hypothesis, revived. AB - Recent evidence shows that the relation between birth order and intelligence is not the same in cross-sectional and within-family data. This simple empirical observation invalidates the conclusions from hundreds of previous birth order studies that relied on cross-sectional data. Simultaneously, the empirical foundation disappears from underneath theories like dilution and the confluence model that use explanatory processes occurring within the family. A theory proposed almost 25 years ago--the admixture hypothesis--effectively accounts for these empirical patterns. In this article, the author describes why birth order is of such intense interest to both parents and researchers (the birth order trap), discusses past birth order-intelligence patterns, shows that the admixture hypothesis accounts for those patterns, and reframes the original argument to support future productive research efforts. PMID- 11413875 TI - Herbert A. Simon (1916-2001). PMID- 11413876 TI - Methodology, birth order, intelligence, and personality. PMID- 11413877 TI - On family size and intelligence. PMID- 11413878 TI - Birth order and intelligence: together again for the last time? PMID- 11413879 TI - Biliary atresia at the dawn of a new century. AB - Biliary atresia is a unique disorder in pediatric surgery. The incidence is 1 in 3,500 to 20,000 live births, but the disorder accounts for more than half of the patients requiring liver transplantation. After Kasai operation, the 5- and 10 year survival rate without the need of transplantation are 37-48% and 18-28%, respectively. An infectious pathogen, particularly the virus, has long been implicated as the causative agent of biliary atresia, but none has ever been identified. Ductal plate malformation is likely to be important in the pathogenesis of biliary atresia. Earlier diagnosis is associated with a better prognosis and biliary atresia should be considered in the differential diagnosis of prolonged jaundice in any infants older than 2 weeks of age. The experience of the surgeon and surgical decade are determinant in achieving success of the Kasai operation. Postoperative cholangitis affects more than 45% of the patients receiving Kasai operation and the afflicted patients are at risk of cessation of bile flow and exacerbation of preexisting liver cirrhosis. Hepatic fibrosis starts at the moment when biliary atresia develops and the ongoing inflammation will end up with liver cirrhosis and hepatic failure in most patients, including some with adequate bile drainage. The inhibition of liver cirrhosis in mice or rats by gene therapy, such as telomere gene delivery, transduction with the hepatocyte growth factor gene or blockade of TGF-beta signaling, may provide a new strategy to rejuvenate the ailing liver associated with biliary atresia in the future. PMID- 11413880 TI - Craniofacial surgical simulation: application of three-dimensional medical imaging and rapid prototyping models. AB - BACKGROUND: Adequate assessment of the deformity, formulation of good treatment planning, and sufficient rehearsal of procedures before actually performing surgery ensure successful craniofacial surgery. Three-dimensional computed tomography (CT) imaging and facsimile models were used in combination to evaluate their function in craniofacial surgery. METHODS: Three-dimensional imaging and facsimile models were used for reconstruction of craniofacial deformity. CT data were acquired, processed, and reconstructed to display 3-dimensional images. The images were used for evaluation of the deformity. The images were then manipulated to create multiple osseous objects. Surgical simulation was performed by moving the computer images. The image processing and manipulation were achieved using the AnalyzePC program. The raw CT data were transformed into a readable format and transferred to produce facsimile models using rapid prototyping technology. The skull models were used for evaluation and surgical simulation. Both methods were compared and used to assist in surgery, which was performed according to the simulations. RESULTS: Three-dimensional CT imaging and facsimile models were helpful for simulation of craniofacial surgery. The actual surgery results were satisfactory without complications. Particular advantages were the unlimited trials with the imaging method, and the feeling of reality with the model method. CONCLUSIONS: Craniofacial surgery is facilitated by preoperative simulation of procedures. Both 3-dimensional CT imaging and facsimile models are helpful for craniofacial surgical simulation. PMID- 11413881 TI - Study of plasma endothelin-1 concentrations in Taiwanese neonates with respiratory distress. AB - BACKGROUND: Endothelin-1 (ET-1) is a novel and potent endothelium-derived vasoconstriction peptide present in human plasma. In this study, plasma ET-1 concentrations were determined and their physiological significance was evaluated in Taiwanese neonates with respiratory distress. METHODS: Sixty newborn infants consisting of 22 with respiratory distress syndrome (RDS), 13 with transient tachypnea of newborn (TTNB), 4 with meconium aspiration syndrome, 10 healthy preterm and 11 healthy full-term infants were included for plasma ET-1 determination. Plasma ET-1 levels were measured by enzyme immunoassay at the of age of 1 day. For those who were diagnosed with RDS, plasma ET-1 concentrations were scheduled for evaluation at the ages of 1, 2, 3, 7, 14, 28, and 35 days as long as oxygen was being used. RESULTS: On the first day of life, there was no significant difference in plasma ET-1 concentrations between healthy preterm and term infants (3.92 +/- 0.88 vs. 3.56 +/- 1.98 pg/mL, p = 0.606). However, plasma ET-1 concentrations of infants with RDS were significantly higher than those with TTNB (6.46 +/- 0.58 vs. 3.77 +/- 1.29 pg/mL, p < 0.001). In RDS infants, plasma ET-1 concentrations showed no significant difference between those who developed bronchopulmonary dysplasia (BPD, N = 4) and those who recovered (non-BPD, n = 18) (7.84 +/- 1.85 vs. 5.81 +/- 2.76 pg/mL, p = 0.242). CONCLUSION: Plasma ET-1 concentrations were similar in preterm and term infants. ET-1 concentrations were higher in infants with RDS than in infants with TTNB, which suggests that plasma ET-1 levels can be useful in the differential diagnosis. However, the plasma ET-1 concentrations can not be a predictor for BPD. PMID- 11413882 TI - Acute appendicitis during pregnancy: a clinical assessment. AB - BACKGROUND: Acute appendicitis is the most common non-obstetric reason for laparotomy during pregnancy. The purpose of this study was to analyze the characteristics of the clinical presentations and postoperative outcomes of these patients and their fetuses. METHODS: Patients who underwent appendectomies during pregnancy from July 1991 to June 1997 were retrospectively identified. Their ages, clinical presentations, the severity of the inflammatory change in the appendices, and the postoperative complications of these patients and fetuses were recorded and analyzed. Long-term outcomes were confirmed by telephone contact, when possible. RESULTS: Forty-five pregnant women who underwent appendectomies for suspected acute appendicitis were retrospectively reviewed. The histopathological inflammatory change in the appendix was proven in 35 patients (78%). Sixty-three percent of the patients were multiparous, and 86% were in the first 2 trimesters. In the clinical settings, pain and tenderness in the right lower abdominal quadrant were the most common symptoms and signs in presentation. Perioperative administration of ritodrine had no obvious advantage in the prevention of fetal loss. Appendectomy was performed beyond 36 hours of onset of symptoms in 28% of the patients, among whom one-half had gangrenous or perforated appendices. Only one fetus was spontaneously lost in 32 patients with diseased appendices (3%), excluding 3 patients choosing artificial abortion. There was no maternal death in our series. CONCLUSION: The rate of fetal loss due to surgery for acute appendicitis during pregnancy was low. Delay of operation was pertinent to the more-inflammatory changes of the appendix and to the higher maternal complication rate. Early surgical intervention is essential. PMID- 11413883 TI - Effect of methylglyoxal on mouse embryogenesis. AB - BACKGROUND: Methylglyoxal (MG), a highly active and mutagenetic compound, has been found widely in a variety of foods and beverages. We investigated the effect of MG on mouse embryo development in-vitro. METHODS: Two-cell mouse embryos were divided into six groups according to the MG concentration in the culture medium: Group 1 (control group). 0 mM; Group 2, 10(-4) mM; Group 3, 10(-3) mM; Group 4, 10(-2) mM; Group 5, 10(-1) mM; Group 6, 1 mM. Embryo development and cleavage were compared every day for 5 days. RESULTS: The percentages of embryos reaching blastocyst/hatching stages were as follows: Group 1, 66.8%/34.2%; Group 2, 67.9%/38.7%; Group 3, 56.2%/31.5%; Group 4, 39.4%/14.1%; Group 5, 11.4%/10.2%; Group 6, 0%/0%. Higher MG concentrations (> or = 10(-2) mM) were associated with morphological aberrations and blocked development of embryos. CONCLUSION: The cutoff value of MG concentration on the mouse embryo development in-vitro is 10( 2) mM. An increased risk of embryotoxicity occurs with MG concentrations > or = 10(-2) mM in vitro. There were no significant effects on the growth rate at MG concentrations of 10(-3) and 10(-4) mM. PMID- 11413884 TI - Phenotype II triploid pregnancy and study of the parental origin of the extra set of chromosomes with fluorescence microsatellite analysis: case report. AB - Triploidy can reflect diandry (dispermy or diploid sperm) or digyny(diploid ovum). The former is likely to result in the type I phenotype with a partial mole with an appropriate-for-gestational age fetus rarely surviving beyond 20 weeks and a large, cystic placenta. The latter, however, is characterized by a type II phenotype with severe intrauterine growth retardation (IUGR) with longer in utero survival and a small, non-molar placenta. We report on a 22-year-old woman, gravida 2, para1, in the 31st week of gestation who was referred to our prenatal clinic for evaluation of severe IUGR and oligohydramnios. Late cytogenetic karyotyping from cordocentesis revealed a triploidy of 69, XXX. In the 33rd week of gestation, the mother went through spontaneous labor delivering an IUGR 1180 gm fetus and a small, non-molar placenta. The fetus died immediately and was sent for autopsy. In addition to cleft lip and palate, the infant had pulmonary lobation abnormalities. Fluorescence microsatellite analysis of fetal and parental samples confirmed that the extra set of chromosomes present in the proband was a result of a maternal meiosis I nondisjunction error. This may help study of genomic imprinting on human development. PMID- 11413885 TI - Idiopathic polypoidal choroidal vasculopathy: case report. AB - In 1990, Yannuzzi et al. introduced a new clinical entity, "idiopathic polypoidal choroidal vasculopathy" (IPCV). There is now a more-widespread worldwide recognition of a higher prevalence and importance of this disorder. We report on the first case of IPCV in Taiwan to our knowledge. A 63-year-old Taiwanese man complained of blurred vision in his right eye for 6 months. The anterior segment and vitreous were normal except for mild nuclear cataract changes. Fundus examination of the right eye showed hard exudates and multiple reddish-orange subretinal lesions in the macula. Fundus examination of the left eye was normal. Fluorescein angiography of the right eye showed multiple hyperfluorescent granular lesions in the macula. Subretinal leakage and pooling of fluorescein were present in the temporal macula. Indocyanine green angiography disclosed multiple aneurysmal dilations at the terminals of branching vascular networks. A hyperfluorescent aneurysmal lesion corresponding to the reddish-orange nodule was observed ophthalmoscopically, and a granular lesion was observed in fluorescein angiography. Focal retinal photocoagulation was applied to the leakage area identified with fluorescein angiography. Two months after photocoagulation was performed, the subretinal fluid had gradually resolved, but a new leakage in the macula was demonstrated with fluorescein angiography. His best corrected vision in the right eye was stable. PMID- 11413886 TI - Osteoblastoma of the patella: case report. AB - A 34-year-old man came to our clinic because of left knee pain and mild swelling. Local tenderness over the peripatellar area was observed during physical examination. The range of motion of the knee was full. The radiographic presentation of the patella revealed an osteolytic lesion with a thin sclerotic rim without evidence of extra-articular involvement. The computed tomography revealed an upper pole intraosseous lesion. Intralesional curettage and allogeneic bone grafting were carried out. The pathologic examination demonstrated primitive osteoblasts, with osteoid features and many giant cells. A diagnosis of the osteoblastoma was made. Following surgery, the patient regained full range of motion and was pain-free. Complete healing of the lesion without evidence of recurrence was noted in follow-up radiographs 2 years postoperatively. PMID- 11413887 TI - Bilateral diffuse uveal melanocytic proliferation in a woman with uterine leiomyoma: case report. AB - Bilateral diffuse uveal melanocytic proliferation (BDUMP) associated with systemic carcinoma is a rare paraneoplastic syndrome. It was reported to be associated with primary carcinomas of the lung, ovary, colon, gall bladder, esophagus, pancreas, uterus, and cervix. We report a case of BDUMP that occurred in a woman with cervical leiomyoma. The 79-year-old woman experienced bilateral progressive visual loss to hand motion within 2 weeks. Fundus examination showed bilateral exudative retinal detachment and multiple oval red patches. Echographic study showed diffuse thickening of the uveal tracts. Fluorescein angiography showed multiple patches of window defect without late leakage, which was correlated with the oval red patches. After screening for tumor markers and meticulous examinations, an elevated serum level of CA-125 was noted. The only neoplasm found in this patient was a cervical leiomyoma, which has never been reported to be associated with BDUMP. One month after the onset of symptoms, accidental head injury due to poor vision led to right parietal intracerebral hemorrhage and left hemiplegia. Prompt diagnosis of this paraneoplastic syndrome is important to discover the underlying malignancy in patients with no history of cancer. For those cases without malignant neoplasm, attention should be directed to the disability caused by the severe visual loss, which led to accidental head injury and intracerebral hemorrhage in this case. PMID- 11413888 TI - [Music therapy in Alzheimer's disease: is an evidence-based approach possible?]. AB - The application of music therapy to Alzheimer's patients is relatively recent. The studies available in literature show that music therapy has a positive effect either on mood or cognition. However, the generalization of data is difficult because these researches have lacked methodological design rigour. Based on the application of this rehabilitative technique in our Alzheimer Unit in Brescia and on the recent researches of music neuroanatomy, this work tries to identify which processes are involved in the therapeutic effect of music therapy. Despite the perceived effect on mood and socialisation abilities, cultural and methodological problems hinder to demonstrate the efficacy of music therapy. Information about neurophysiological and neurochemical correlates of music therapy are so poor that the use of this technique is often based on the assumption that the supposed positive effect of music is enough to justify its application. The methodological problem is related to the evaluation of outcomes. The fact that those studies which investigated the effects of music therapy were characterized by less specific indicators (cognition, behavior) and by less standardized instruments made difficult to generalize and quantify the results. The aim of the study is to organize the present knowledge with a systematic approach so that further researches lead to base the application of music therapy on evidence instead of on singular clinical finding. PMID- 11413889 TI - [Hematopoietic stem cell transplantation after nonmyeloablative conditioning]. AB - In these last years it has become evident that nonmyeloablative stem cell transplantation (NST) is a feasible procedure at least in the first 100 days. Generally NST has less severe adverse effects than conventional allografting. Unfortunately, short observation period cannot permit us to give a definitive reply to the question if this procedure will substitute the conventional myeloablative transplantation. NST is a promising new option for the treatment of hematologic and non-hematologic neoplasia, which role will be better defined in the near future. PMID- 11413890 TI - [Assessment of cognitive and functional status in hospitalized elderly]. AB - In 1745 elderly patients (971 men and 774 women), consecutively admitted to University Geriatric Ward in San Giovanni Battista Hospital of Torino, cognitive status (SPMSQ) was evaluated. Moderate-severe and mild cognitive impairment were respectively present in 26.8% and 15.8% of the sample. Subjects affected by moderate-severe cognitive impairment were older (80.7 +/- 8.0 yrs) than patients with mild deterioration (79.5 +/- 7.8 yrs) and patients without cognitive problems (75.3 +/- 8.3 yrs). The number of pathologies and the length of hospital stay were not related to severe cognitive impairment. Lower schooling was associated with cognitive alteration. Among subjects affected by moderate-severe cognitive deterioration 15.6% was living with assistance before hospitalization. The prevalence of functional impairment (ADL) was higher (70.9%) among patients with moderate-severe cognitive impairment. In-hospital mortality was higher (21.2%) among patients with moderate-severe cognitive deterioration. PMID- 11413891 TI - [Diagnosis of Helicobacter pylori infection: comparison of techniques]. AB - Several diagnostic assays are available for evaluating Helicobacter pylori infection: histological examination, culture of gastric biopsies, urea breath test and serology. Recently a new enzyme immunoassay has been introduced for the detection of H. pylori antigens in stool samples (HpSA). The aim of our study was to evaluate and compare diagnostic efficacy of HpSA with histological examination, culture, urea breath test and serology in a group of 95 patients. Patients were classified H. pylori positive (43) or negative (52) on the basis of histology, culture and urea breath test. HpSA optical densities were significantly higher in infected patients compared to those obtained in H. pylori negative patients (t = 5.47, p < 0.001). Overall, with a fixed cut-off of 0.1 unit of optical density, the sensitivity was 79% and the specificity 100%. In the H. pylori positive patients, HpSA optical density correlated with bacterial load histologically evaluated in the gastric antrum (r = 0.405, p < 0.05) and was inverse correlated with levels of serum IgG elicited against H. pylori (r = 0.315, p < 0.05). Considering patients with a positive HpSA finding and/or levels of anti-H. pylori antibodies upper than 30 U/mL, sensitivity in detecting infected patients was 98%. IN CONCLUSION: (1) immunodetection of H. pylori antigens in stools is a good alternative of breath test; (2) a reduction in H. pylori density grade might be accompanied by low HpSA optical density, leading to a false negative result and (3) combining the HpSA determination with the serum detection of anti-H. pylori antibodies a better clinical sensitivity is obtained. PMID- 11413892 TI - [Interstitial pulmonary fibrosis in chronic HCV hepatitis treated with interferon]. AB - The study describes the clinical, virologic and immunological characteristics of a patient with interstitial pulmonary fibrosis during interferon treatment for HCV chronic hepatitis. After the interruption of the interferon and the beginning of immunosuppressive treatment, an improvement of pulmonary pathology was observed. At the reintroduction of interferon, the patient presented a rapid worsening of pulmonary fibrosis with a normalization of biochemical and virologic parameters of hepatitis. The correlation among interstitial pulmonary fibrosis, HCV infection and interferon treatment is discussed; however in the described case, the pulmonary pathology was correlated to interferon treatment. PMID- 11413893 TI - [Pharmacologic therapy in the elderly]. AB - The pharmacological treatment in the older people should be carefully considered: older patients are at the same time the greatest consumers of drugs and the population with the greatest risk of adverse drug reactions (ADR). The ADR are in the old patient more frequent and serious for the greater number of concurrent drugs. The incorrect drug's use consists in prescribing too much or too little, for an excessive period, without a defined diagnosis and an appropriate selection of the better compound. Moreover, beneficial drugs may be frequent underused, some chronic conditions do not receive adequate pharmacologic treatment, and an ADR may be misinterpreted as a new pathological condition that requires a new prescription. The unusual presentation of many diseases in elderly play a role to complicate the clinical process necessary to optimising the drug treatment. About the risks and the benefits of pharmacological treatment, adequate data in older patients are in general lacking for the poor inclusion in clinical trials of the elderly subjects, especially frail persons. PMID- 11413894 TI - [Arterial hypertension in very old people. Problems of treatment]. AB - Several epidemiological tests showed that, in the very old, low blood pressure was associated with higher mortality and hypertension with a longer life. But more recent studies, some from our school, and the data from Framingham have clarified that the higher mortality in the very old with low blood pressure is to be attributed to pre-existing pathologies which caused the low blood pressure in the first place. Though specific intervention trials on the very old are not available, the analyses of the results of antihypertensive therapy in the sub groups of the very old in some trials carried-out with elderly hypertensives suggest the use of pharmacological treatment also in the very old (> 80 yrs), at least for isolated systolic hypertension. The therapy must be based on a careful choice of drugs and carried-out with particular caution to avoid disability from hypertension as well as to avoid side effects which worsen the quality of life of your very old patients. PMID- 11413895 TI - [Acute respiratory failure in organ transplantation and immunodepressed patients]. AB - Recently, noninvasive ventilation (NIV) has been introduced as a new and alternative ventilatory method. The efficacy of NIV has been reported in immunosuppressed patients, with the decrease in drawbacks related to mechanical ventilation by endotracheal intubation. The authors propose an up-date on the use of NIV in transplanted patients. PMID- 11413896 TI - [Periodic clinical monitoring after liver transplantation]. AB - During the last two decades, owing to advances in immunosuppressive pharmacotherapy, liver transplantation has been increasingly accepted by the medical community as an effective treatment for patients with end-stage liver disease. Successful transplantation of the liver, however, requires frequent monitoring. Most of the serious infectious complications and allograft dysfunction occur during the early post-transplantation period (i.e., first six months). Blood levels of cyclosporine or tacrolimus, the two major calcineurin inhibitors currently in use, need to be frequently checked. Drug dosage is adjusted in order to maintain target serum concentrations and the patients free of side-effects. In the time, the risk of acute allograft rejection decreases considerably, whereas the proportion of patients with fibrosis or cirrhosis increases, particularly among hepatitis C virus carriers. Graft loss may occur, secondary to recurrent disease or chronic rejection. Patients with well functioning grafts may still be affected by significant comorbidities, such as hypertension, diabetes, obesity, hyperlipidemia and osteoporosis, which appear to be related to long-term immunosuppression. The incidence of lymphoma, skin and colorectal cancers in liver transplantation recipients exceeds those found in the general population and requires early detection. The principles of the management of medical problems after liver transplantation are a careful clinical assessment of the patient and a judicious use of laboratory tests, radiological evaluation and liver biopsy. PMID- 11413897 TI - [Steroids, cognitive processes and aging]. AB - Adrenal steroids, besides acting on the pituitary and the hypothalamus to exert classical feedback activity, can also have endocrine and extra-endocrine actions at the level of sub-cortical areas, as the hippocampus and the amygdala, involved in cognition and effectiveness. Acting on the hippocampus, an area particularly equipped with specific receptor sites, adrenal steroids exert either a feedback inhibition on their own secretion or a morphological and functional age-related deterioration of this target area. Loss of hippocampal neurons and corticosteroid receptors with ageing endangers the feedback inhibitory action of the steroids, and induces an over-exposition of the hippocampus to their detrimental action, enhancing the vulnerability of the neuronal cells to metabolic stimuli (hypoxia, hypoglycemia). Hence, either in the physiological ageing of the brain as well as in age-related neurological diseases or psychiatric diseases, harboring a primary neuro-anatomical-functional alteration of the hippocampus, or when the hippocampus is over-exposed to adrenal steroids, a cohort of cognitive and behavioral alterations may be partly due to adrenal gland hyperfunction. Gonadal steroids exert effects on the CNS which go far beyond regulation of gonadotropin secretion and sexual function, though their action is opposite to that of adrenal steroids. Estrogens stimulate hippocampal synaptogenesis, enhance cholinergic neurotransmission, possess anti-oxidative and anti-amiloidogenic properties, dilate cerebral vessels and have platelet anti-aggregating activity. One is led to postulate that the dramatic decrease of estrogen secretion at menopause should increase the vulnerability of the CNS by the many factors contributing to neurodegeneration associated to brain ageing or Alzheimer disease. PMID- 11413899 TI - Introduction. Approach to the hospitalized asthma patient. PMID- 11413900 TI - Status asthmaticus. From the emergency department to the intensive care unit. PMID- 11413901 TI - Pharmacologic management of the hospitalized pediatric asthma patient. PMID- 11413902 TI - Asthma in the hospitalized obstetrical patient. PMID- 11413903 TI - Discharge of the asthmatic patient. AB - Asthma continues to be a challenging disease to treat in both the inpatient and outpatient settings. The growing database on therapeutic interventions at the time of transition from the acute to chronic phase of this disease is encouraging. Glucocorticoids and inhaled beta-agonists clearly reduce readmission and relapse. Other medications and educational interventions also appear effective. Still, no true discharge guidelines have been established. Multiple statements by consensus panels have recommended using FEV1 or PEFR as indicators of readiness for discharge, but this has not been prospectively validated from either the emergency department or inpatient setting. In contrast, some studies argue that pulmonary functions do not accurately predict relapse and readmission, so the usefulness of these discharge recommendations is debatable. Large studies, especially in the adult asthmatic population, are needed to validate these recommendation. PMID- 11413904 TI - Pharmacologic treatment of the adult hospitalized asthma patient. AB - Acute severe asthma calls for aggressive and early therapy of a multifaceted and all-inclusive approach (Fig. 2). Therapy merely begins in the ED and manifold distinct issues need to find consideration during ongoing hospital care. Currently, beta-agonists, anti-cholinergic agents, and corticosteroids remain the mainstay of therapy. Methylxanthines and magnesium may find consideration in carefully selected patients. Multiple new therapeutic avenues, such as the anti leukotriene drugs, seem promising and future studies will hopefully extend our armamentarium against life threatening complication of a common disease. Asthma education begun in the hospital may provide the platform for preventing severe acute exacerbations and hospitalization. PMID- 11413905 TI - Mechanical ventilation in severe asthma. PMID- 11413906 TI - When conventional asthma therapies fail. PMID- 11413907 TI - [Difficulties in noninvasive cardiac stroke volume determination in "small" blood circulation systems]. AB - Pulsewave velocity analysis is an option for the noninvasive determination of cardiac output and the evaluation of additional haemodynamic parameters. An algorithm we developed ourselves has been established as a method for the measurement of cardiac output in rabbits. The effectiveness of this program was investigated by monitoring controlled hypoxia-induced alterations in the circulatory system. Calculated values were compared with direct measurements of cardiac output with a Doppler flow probe placed in the ascending aorta. Within the physiological framework of blood gas analysis, a good correlation was found between the two methods. In the case of hypoxia-induced depression, however, the two methods showed diverging results, presumably due to arrhythmia-induced wave reflections within the arterial vascular tree. This makes an extension of the algorithm necessary to take account for these case. PMID- 11413908 TI - [Hemodynamics of the lower extremity with pneumatic foot compression. Effect on leg position]. AB - External pneumatic compression of the foot is being used more and more to increase the venous blood flow in the lower limbs and thus reduce the risk of postoperative deep venous thrombosis. We have investigated the efficacy of the foot pump pneumatic compression device (A-V Impulse, Novamedix, Andover, England) in 10 healthy subjects and in 10 patients undergoing total hip arthroplasty. The velocity of venous blood flow in the common femoral artery was measured in the horizontal, Trendelenburg (head-down, foot-up) and reverse Trendelenburg (head-up and foot-down) positions using a duplex ultrasound unit (Sonoline Elegra, Siemens, Erlangen, Germany) with a 5 MHz linear array probe. Application of the foot pump produced an increase in venous blood flow velocity in all healthy subjects and in all patients. In healthy subjects the mean increase in the horizontal position was 31.18% (SD = 15.86%), and in the Trendelenburg position 20.72 (SD = 15.69%) (right limb). In the reverse Trendelenburg position, the foot pump produced a mean increase of 94.08% (SD = 55.00%). The difference is statistically significant (p < 0.005). In patients with a total hip arthroplasty, the mean increase in the horizontal position was 28.67% (SD = 9.95%), and in the Trendelenburg position 20.34% (SD = 17.85%) (operated limb). In the reverse Trendelenburg, the foot pump produced a mean increase of 91.55% (SD = 42.38%). The difference is statistically significant (p < 0.005). There was no noteworthy difference in results between the controls and patients. Pneumatic compression devices designed to reduce venous stasis are effective in decreasing the rate of deep venous thrombosis after surgery on the lower limb. The results of the present study show that the efficiency of the foot pump in increasing venous return is improved by adopting the reverse Trendelenburg position. This may increase its thromboprophylactic effect. PMID- 11413909 TI - [Artificial intelligence in sleep analysis (ARTISANA)--modelling visual processes in sleep classification]. AB - We describe a novel approach to the problem of automated sleep stage recognition. The ARTISANA algorithm mimics the behaviour of a human expert visually scoring sleep stages (Rechtschaffen and Kales classification). It comprises a number of interacting components that imitate the stepwise approach of the human expert, and artificial intelligence components. On the basis of parameters extracted at 1 s intervals from the signal curves, artificial neural networks recognize the incidence of typical patterns, e.g. delta activity or K complexes. This is followed by a rule interpretation stage that identifies the sleep stage with the aid of a neuro-fuzzy system while taking account of the context. Validation studies based on the records of 8 patients with obstructive sleep apnoea have confirmed the potential of this approach. Further features of the system include the transparency of the decision-taking process, and the flexibility of the option for expanding the system to cover new patterns and criteria. PMID- 11413910 TI - [Development of an incubation system for an inverted microscopy for long-term observation of cell cultures using chamber slides]. AB - Trifunctional bispecific antibodies open up new immunological possibilities in tumour treatment. Prior to clinical application, comprehensive investigations using animal models and in vitro examinations need to be done. To investigate long-term interactions between various immunologically active blood cells and individual tumour cells in the presence of antibodies, we developed an incubation system for experimental cell cultures on an inverted microscope. The system consists of a perspex box with a central moisture chamber with integrated water reservoir, external air circulation heating, and a CO2 supply. The sterile cell cultures are located in the wells of a slide positioned within a depression in the water reservoir. The newly developed incubation system enables continuous observation over the long term of experiments under optimal cell cultures conditions in combination with modern video techniques. PMID- 11413911 TI - Pruritometer 2: portable recording system for the quantification of scratching as objective criterion for the pruritus. AB - Studies to evaluate therapies for itching (pruritus) related diseases often require the quantification of the itch sensation. Like all subjective symptoms the evaluation of itching is difficult and can only be done indirectly. With the Pruritometer 2 a measuring system is introduced that evaluates itching by detecting scratching movements. Based on the Pruritometer 1, that processes the signals of a piezoelectric vibration sensor, fixed on the midfinger of the patients dominant hand, and triggers a simple counter, the Pruritometer 2 allows to store the scratch activity during a 24 hours period. For each adjustable time slice of this time period, the amount of scratches and the scratch intensity are recorded. All data can be transferred to a PC via infrared link for further processing with a standard software package. An additional PC-software allows to set various parameters for optimal scratch detection and to test the patient attached system, also via the infrared link. All electronic components are shockproof encapsulated in a milled housing and are attached to a textile watchstrap that is worn by the patient like a wristwatch. PMID- 11413912 TI - [Results of bicondylar sledge prostheses with special reference to the gait pattern]. AB - AIM: To establish whether the clinical results obtained with bicondylar sledge prostheses correlate with clinical findings, and whether clinical results influence gait pattern. MATERIAL AND METHODS: 15 patients with bicondylar sledge prostheses were compared with healthy controls. The clinical examination was documented using the HSS, Knee Society, Tegner, and patella scores, and the Visual Analogue Scale. Gait analysis was done with a three-dimensional motion analysis system and force platforms. RESULTS: A comparison of patients and controls showed significant differences in the clinical results, whereas the parameters gait function, muscle force and pain did not. No significant differences were found between the operated and non-operated legs. In comparison with controls, patients showed significant differences in the ground reaction forces. DISCUSSION: The abnormal gait findings showed no correlation with clinical results, nor did such parameters as age, weight and duration of follow up appear to have any influence. The non-operated leg adapted to the gait pattern of the operated leg, to restore gait symmetry. CLINICAL RELEVANCE: Subjective complaints or clinical results show little relationship to gait parameters in patients receiving a bicondylar sledge prosthesis. PMID- 11413913 TI - Long-term management and prognosis of autoimmune hepatitis (AIH): a single center experience. AB - BACKGROUND: Controlled trials have firmly established the need for immunosuppressive therapy in autoimmune hepatitis. However, reports about long term management and prognosis of the disease are scarce. PATIENTS AND METHODS: We reviewed the charts of 103 consecutive patients with a well-documented long-term course of autoimmune hepatitis who had been carefully managed over a mean observation period of 95 months (12-405 months). RESULTS: Under immunosuppressive therapy 94 patients (91.2%) reached complete remission after a mean treatment duration of 3 +/- 3 months. 28 of the 103 patients (27.2%) were eligible for a trial of treatment withdrawal after a mean treatment duration of 32.2 months (range: 12-81 months). 21 of these patients (75%) had a relapse following treatment withdrawal. 13.6% of patients had intolerance of or severe side effects to azathioprine. There was no increase in tumor risk during a cumulative observation period of 423 patient-years of azathioprine therapy. Corticosteroid side effects occurred mostly during induction therapy, but were usually minor and resolved upon dose reduction. During a cumulative observation period of 842 patient-years no liver related deaths occurred and no patient had to be referred to liver transplantation, even though 30 patients (29.1%) had histological evidence of cirrhosis at presentation. The overall 5- and 10-year survival of patients with autoimmune hepatitis was identical to an age- and sex-matched control population. CONCLUSION: Our study shows that the majority of patients with AIH do achieve a complete remission within 3 months, but require long-term or permanent immunosuppressive therapy that is usually well tolerated. Long-term survival in well-managed patients is excellent. PMID- 11413914 TI - Improvement in the quality of the endoscopic/bioptic diagnosis of gastric ulcers between 1990 and 1997--an analysis of 1,658 patients. AB - BACKGROUND: Helicobacter pylori research has greatly improved the diagnosis and treatment of gastric ulcer. The aim of the present study was to investigate the question whether the new perceptions had been incorporated into the daily diagnostic routine of the nineteen-nineties. PATIENTS AND METHODS: Analysis of the biopsy material at the institute of Pathology of the Klinikum Bayreuth obtained from 1,658 patients with gastric ulcers in the months of January and February 1990 (n = 506), 1992 (n = 525), 1994 (n = 532), 1995 (n = 432) and 1997 (n = 400) with the aim of determining the quality of the endoscopic assessment of biological significance, and the number of biopsy specimens obtained from the ulcer, as well as the number of additional biopsies taken from the antrum and corpus. RESULTS: 1,436 (88.9%) of the endoscopically diagnosed ulcers proved to be histologically benign and 222 (11.1%) malignant, of which 192 (86.5%) were carcinomas, 24 (10.8%) MALT lymphomas, 2 (0.9%) sarcomas, and 4 (1.8%) ulcerated metastases. The percentage of false-negative endoscopic assessments of biological significance was 1.8%, that of the false-positive assessments 4.1%, that is, both very low figures. In the case of endoscopically suspicious findings, the histological examination revealed an ulcerated malignant tumor in almost 30% of the cases. The average number of biopsies harvested from the ulcer increased from 5.9 in 1990 to 7.1 in 1997. The frequency of biopsies obtained additionally from antrum and corpus rose statistically significantly from 14.1% of the cases in 1990 to 43.0% in 1997. CONCLUSION: During the course of the nineteen-nineties, the quality of the endoscopic/bioptic diagnosis improved noticeably. In terms of the number of biopsy specimens obtained from the ulcer to identify its biological significance, and the number of additional biopsies from antrum and corpus to identify underlying diseases, there is, however, still room for improvement. PMID- 11413915 TI - [Diagnosis related groups (DRG)--current significance in gastroenterology]. AB - Prospective payment systems using the diagnosis related groups (DRG) payment mechanism have been introduced in many countries all over the world. In June 2000 the Australian Refined Diagnosis Related Groups System (AR-DRG) was chosen as the model which will be introduced in 2003 as the control instrument for the hospital dependent public health system in Germany. Background and possible implications for gastroenterologists and hepatologists are discussed. PMID- 11413916 TI - [Colorectal carcinoma in Cronkhite-Canada syndrome]. AB - We report a 63-year-old lady with Cronkhite-Canada syndrome, who developed colorectal cancer. A hemicolectomy was performed, and the tumor specimen was prepared for DNA-analysis and immunohistochemical screening. We found a mutation of p53 gene without APC- and ras-gene alteration and expression of erbB2 protooncogen. The polyps in non-hereditary Cronkhite-Canada-syndrom are neither adenomatous nor hyperplastic, but patients often develop colorectal cancers. The steps of mutation do not follow the adenoma-carcinoma sequence, first described by Vogelstein 1988. This and previous observations suggest that carcinogenesis in Cronkhite-Canada syndrome follows another independent sequence. PMID- 11413917 TI - [Differential mucin expression in the gastrointestinal tract]. AB - Under physiological conditions mucins display a tissue specific expression. MUC2, MUC3, MUC5AC and MUC6 are the major mucins in the gastrointestinal tract. In the intestinal tissue MUC2 and MUC3 are the predominant mucins, whereas MUC5AC and MUC6 are the most important mucins in the stomach. Pathophysiological conditions are characterized by an aberrant gene expression. This includes the expression of non-tissue specific mucins as well as a reduced expression of the specific mucins. During inflammation a co-expression of non-tissue specific mucins was observed. Neoplastic transformations are also associated with an aberrant mucin expression. Obviously, an increased expression of non-specific mucins is related with a more favorable prognosis, while a decreased mucin expression is indicative for an increased cellular dedifferentiation and a poor prognosis. This modified mucin expression may result from a modified gene expression as well as from modifications in the posttranscriptional processing. A modified mucin expression reflects pathophysiological conditions and may support the pathohistological diagnosis. PMID- 11413918 TI - [Is induction of oral tolerance to to insulin a suitable treatment concept in therapy of type I diabetes mellitus?]. PMID- 11413919 TI - [Pathogenesis of non-alcoholic fatty liver]. PMID- 11413920 TI - [The selective Cox-2 inhibition by rofecoxib reduces risk of severe gastrointestinal complications of anti-inflammatory therapy by more than 50%]. PMID- 11413921 TI - Antifungal activities of pelargonic and capric acid on Microsporum gypseum. AB - The inhibitory effects of pelargonic and capric acid on Microsporum gypseum were examined. Solid and liquid Sabouraud glucose media containing different concentrations of pelargonic and capric acid were separately prepared and inoculated with the suspension of mycelium and spores of M. gypseum and incubated at 25 degrees C for 1 month. The culture media were examined periodically for fungal growth and the minimum inhibitory concentration (MIC) of each fatty acid was determined. The MIC for capric acid was 0.02 mg ml-1 and for pelargonic acid 0.04 mg ml-1 on solid media and 0.075 mg ml-1 for capric acid and 0.05 mg ml-1 for pelargonic acid in the liquid media. PMID- 11413922 TI - Case report. Disseminated cutaneous cryptococcosis in an immunocompetent host. AB - A 50-year-old Indian housewife presented with multiple cutaneous lesions over the upper back, thigh and both the extremities, which first appeared 5 months previously. The diagnosis of disseminated cutaneous cryptococcosis was made on the basis of the demonstration and isolation of Cryptococcus neoformans from the aspirated pus and skin biopsy. A serological test for HIV antibodies was negative. The patient responded to itraconazole treatment. PMID- 11413923 TI - Case report. Cryptococcal cellulitis showing necrotizing vasculitis. AB - A 65-year-old woman with refractory anaemia who had been treated with systemic corticosteroids for several months developed cryptococcal cellulitis of the right cubital fossa. She was treated empirically for a presumed bacterial cellulitis with little response. Histological examination of debrided tissue revealed Cryptococcus as the causative organism. The tissue reaction involved suppurative inflammation with abscess formation in the dermis and subcutaneous adipose tissue. Necrotizing vasculitis, which has rarely been described in cryptococcosis, was seen in this case. Although the cellulitis was cured by local treatment in this patient, most previous reports recommended systemic antifungal therapy to treat cryptococcal cellulitis. PMID- 11413924 TI - Case report. Onychomycosis due to Microsporum canis. AB - A case of distal subungual onychomycosis of the big toe due to Microsporum canis is reported in a 69-year-old male asthma patient who had been treated with systemic corticosteroids for the last 3 years. The nail infection was contracted from a cat who was a healthy carrier. The patient was treated successfully with intermittent itraconazole therapy. PMID- 11413925 TI - Phenotypic and genotypic characterization of reference strains of the genus Aspergillus. AB - Twenty-five culture collection strains from four Aspergillus species (A. fumigatus n = 8, A. flavus n = 8, A. niger n = 4, A. nidulans n = 5) were characterized by four methods: (i) determination of patterns in an assimilation assay; (ii) protein pattern of whole mycelial cell lysates in the sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis (PAGE); (iii) reactivity of a pool serum obtained from cystic fibrosis patients with mycelial lysates in the immunoblot; and (iv) random amplification of polymorphic DNA (RAPD) with eight primers having arbitrary or repetitive sequences. In the assimilation assay the A. fumigatus strains showed identical patterns in contrast to the strains of the species A. flavus, A. niger, and A. nidulans, which each showed four patterns. In the SDS-PAGE no differences in the band patterns in the A. fumigatus strains were found, in contrast to the A. flavus (three patterns), A. nidulans (five patterns) and A. niger strains (two patterns). The immunoblot patterns were characteristic for each species with bands at 62 and 17/18 kDa in the A. fumigatus strains, at 51 and 18 kDa in the A. flavus strains, at 51 kDa in the A. niger strains, and at 51, 40 and 17/18 kDa in the A. nidulans strains allowing, however, no intraspecies typing. In the RAPD assay four out of eight primers gave interpretable patterns with 3-20 bands. None of the primers showed sufficient discriminatory power when used alone. However, when combining the results of two of the primers (5'-GTA TTG CCC T-3' and 5'-GAT AGA TAG ATA GAT A-3') all strains except two A. fumigatus strains could be clearly separated from each other. It is concluded that the the RAPD assay showed the most discriminatory power in all Aspergillus species investigated. In contrast to the phenotypically similar A. fumigatus strains, the strains of the species A. flavus, A. nidulans and A. niger differed in their phenotypic characteristics. The presented data of strains from international culture collections may serve as basis for interlaboratory standardization of typing methods. PMID- 11413926 TI - Onychomycosis caused by Alternaria spp. in Tuscany, Italy from 1985 to 1999. AB - Cutaneous phaeohyphomycosis due to Alternaria spp. is reported with increasing frequency, especially in patients with immune deficiency. Onychomycosis caused by this mould is still rarely observed. Here we report nine cases observed in Tuscany in the period 1985-99; the agent was Alternaria alternata in eight cases and Alternaria chlamidospora in one. Diagnosis was made on the basis of repeated direct microscopic mycological examination and culture, confirmed by scanning electron microscope observation of fragments of colonies. In most cases, the clinical manifestations were dystrophy and distal subungual hyperkeratosis of one or two nails of the feet or hands. Seven cases were treated with oral itraconazole, successfully in six cases, as clinical and mycological recovery was confirmed at follow-up 1 year later. PMID- 11413927 TI - Incidence of Candida in psoriasis--a study on the fungal flora of psoriatic patients. AB - The presence of Candida albicans and other Candida species in saliva and faeces of 50 psoriatic patients compared with a control group of 50 healthy donors was examined quantitatively. The quantity of Candida in saliva and faeces of the psoriatics proved to be significantly higher than in the controls. Candida was detected in 78% of the saliva samples of the psoriatics but in only 50% of the controls, and in the faeces samples in 72% of the psoriatics, but in only 46% of the controls. Qualitative analysis revealed a predominance of Candida albicans (saliva, 77%; faeces, 64%) and Candida rugosa (saliva, 28%; faeces, 28%). We did not find a correlation between the severity of the psoriasis according to the Psoriasis Area and Severity Index and the amount of Candida in the saliva or in the faeces. Our results reinforce the hypothesis that C. albicans is one of the triggers to both exacerbation and persistence of psoriasis. We propose that in psoriatics with a significant quantity of Candida in faeces, an antifungal treatment should be considered as an adjuvant treatment of psoriasis. PMID- 11413928 TI - Prevalence of dermatophytic skin and nail infections in diabetic patients. AB - To determine whether there is a higher frequency of mycotic infections due to dermatophytes in diabetics, 171 diabetic outpatients and 276 controls were recruited in the period 1997-98. Patients with suspicious lesions underwent mycological examination which was positive in seven diabetics and 17 controls. In diabetics the most frequent infection was tinea pedis, followed by distal subungual onychomycosis; the most frequently isolated fungus was Trichophyton mentagrophytes. The results of the study did not show a prevalence of dermatophyte infections in diabetics. No correlation was found between dermatophytosis and duration or type of diabetes and its complications, blood sugar levels or levels of glycosylated haemoglobin. None of the diabetic patients with dermatophytosis had complications related to diabetes and basal blood sugar and glycosylated haemoglobin levels indicated good metabolic control. PMID- 11413929 TI - One week pulse therapy with itraconazole (200 mg day-1) for onychomycosis. Evaluation of treatment results according to patient background. AB - We investigated background factors of patients that affect the response of onycomycosis to pulse therapy with itraconazole. The regimen used in this study involved administering 200 mg of itraconazole orally on a daily basis for 1 week as one pulse, which is half of the normally recommended dose. The number of pulses was fixed at one per month, and altered in accordance with improvements in the infected nails and/or the patients' request. Patient background (n = 63) including sex, age, occupation, duration of the disease, site of lesion (fingernail, toenail), number of affected nails, clinical types, severity of thickening, presence or absence of pathogens, the presence or absence of underlying diseases such as adult onset diseases, turbidity of the target nails before therapy and the number and duration of pulses was investigated. The efficacy of the therapy was evaluated after 12 months. A complete cure was judged to have taken place if an entirely healthy nail plate re-grew in place of the diseased nail, whereas lack of success was defined as the persistence of lesions beyond the designated time period. The average number and duration of pulses were 4.7 +/- 3.2 times over 5.6 +/- 4.3 months, respectively. The rate of complete cure was 62%. Factors in the patients' background that were judged to have an influence on treatment results were the relative duration of the disease, the number of affected nails, and severity of thickening. With regard to the efficacy of the therapy in terms of the number and duration of pulses, 78% of 23 patients who were given pulse therapy two to six times during a course of 2 to 6 months achieved total cure. PMID- 11413930 TI - Effectiveness of terbinafine in the eradication of Microsporum canis from laboratory cats. AB - In this study, the coats of 20 cats with no dermatophytosis lesions were sampled for colonization by Microsporum canis, which was isolated in 45% of cases. All cats were treated with oral terbinafine for 3 weeks. The drug was administered orally at a daily dosage 8.25 mg kg-1. Mycologic status was assessed at the end of the treatment; and after follow-up at 2 days, and 4 and 9 weeks. The final evaluation revealed a complete mycologic cure of all cats by obtaining negative cultures, thereby demonstrating that terbinafine is an effective method of eradication of dermatophyte colonization. PMID- 11413931 TI - In-vitro and in-vivo anti-Trichophyton activity of essential oils by vapour contact. AB - The minimum inhibitory doses (MIDs) of essential oils by vapour contact to inhibit the growth of Trichophyton mentagrophytes and Trichophyton rubrum on agar medium were determined using airtight boxes. Among seven essential oils examined, cinnamon bark oil showed the least MID, followed by lemongrass, thyme and perilla oils. Lavender and tea tree oils showed moderate MID, and citron oil showed the highest MID, being 320 times higher than that of cinnamon bark oil. The MID values were less than the minimum inhibitory concentration (MIC) values determined by agar dilution assay. Furthermore, the minimum agar concentration (MAC) of essential oils absorbed from vapour was determined at the time of MID determination as the second antifungal measure. The MAC value by vapour contact was 1.4 to 4.7 times less than the MAC remaining in the agar at the time of MIC determination by agar dilution assay. Using selected essential oils, the anti Trichophyton activity by vapour contact was examined in more detail. Lemongrass, thyme and perilla oils killed the conidia, inhibited germination and hyphal elongation at 1-4 micrograms ml-1 air, whereas lavender oil was effective at 40 160 micrograms ml-1 air. The in-vivo efficacy of thyme and perilla oils by vapour contact was shown against an experimental tinea pedis in guinea pigs infected with T. mentagrophytes. These results indicated potent anti-Trichophyton action of essential oils by vapour contact. PMID- 11413932 TI - Clinical and echocardiographic features of hypertrophic cardiomyopathy in the elderly. AB - Hypertrophic cardiomyopathy is a familial cardiac disease with exceptionally diverse clinical and morphologic presentations. The influence of age on the disease manifestation has become increasingly clear over the last decade. Most initial reports concentrated on characterization and treatment of the disease in younger individuals, but a better appreciation of hypertrophic cardiomyopathy in elderly patients has yielded important information regarding clinical presentation, morphologic appearance on echocardiography, prognosis, and management. This paper reviews the literature focusing on the age-related differences in hypertrophic cardiomyopathy. PMID- 11413933 TI - Left ventricular diastolic function in the elderly. AB - Previous studies using pulsed Doppler echocardiography have demonstrated a pattern of abnormal left ventricular relaxation associated with increasing age. Specifically, aging is associated with decreased peak velocity of early diastolic mitral inflow, increased peak velocity of late diastolic inflow, increased isovolumic relaxation time, and early diastolic deceleration time. Abnormal relaxation can progress to significantly elevated left atrial pressure- characterized by increased early peak velocity and shortened isovolumic relaxation time and deceleration time--as part of the disease processes. Left ventricular diastolic dysfunction is highly prevalent, occurring in one half to two thirds of elderly patients with congestive heart failure, in association with normal systolic function. Left ventricular hypertrophy, which is commonly related to systemic arterial hypertension, and ischemic heart disease are the two major causes of abnormal left ventricular diastolic function in the elderly. Recently, newer echocardiographic techniques have been described that allow more accurate evaluation of left ventricular diastolic function. Treatments for left ventricular diastolic dysfunction should focus on the underlying disease etiology as well as on the derangement in left ventricular diastolic function. Although calcium channel blockers and angiotensin-converting enzyme inhibitors have been used clinically to treat diastolic dysfunction, their effects on prognosis remain unproven. PMID- 11413934 TI - Transesophageal echocardiographic evaluation of coronary arteries for stenosis in the elderly patient. AB - This article documents the ability of transesophageal echocardiography to provide adequate images and clinically relevant information about the coronary anatomy of the elderly patient. Transesophageal echocardiography is commonly used to assess elderly patients who suffer cerebral vascular accidents. It is important to evaluate not only for the usually suspected causes of a cardiac source of emboli but also for direct and indirect evidence of coronary artery disease--the leading cause of death in the elderly stroke patient. Because atherosclerotic cardiovascular diseases identified in one vascular bed are prone to universally involve the other vascular territories to some degree, it is not surprising that the coronary arteries are often stenotic. As a sudden event with catastrophic symptoms, a stroke is commonly the first vascular event the elderly patient experiences. Depending on the degree of recovery, physical limitations may contribute to the lack of symptoms from coexistent peripheral or coronary artery disease. Transesophageal echocardiography may be the first, or only, coronary evaluation for high-risk elderly patients. PMID- 11413935 TI - Efficacy and safety of angiotensin II receptor blockers in elderly patients with mild to moderate hypertension. AB - The role of the renin-angiotensin-aldosterone system in the pathogenesis of hypertensive disease has long been recognized, and the interruption of this cascade with angiotensin-converting enzyme-I has been beneficial in the management of hypertension. Recently, a new class of drugs, the angiotensin receptor blockers, emerged, enlarging the antihypertensive armamentarium. Since elderly patients are more prone to adverse drug reactions, in this paper we review several trials, most of which were of short duration, on the efficacy and safety of angiotensin receptor blockers in the geriatric population with mild to moderate hypertension. These studies established that the drugs are well tolerated, safe, and in most instances as efficacious as other classes of antihypertensive medications. Combination therapy with angiotensin receptor blockers and hydrochlorothiazide was additive, without any significant effect on the safety profile. PMID- 11413936 TI - Effects of age and gender on the pharmacodynamics of omapatrilat in healthy volunteers. AB - Omapatrilat is the most clinically advanced member of a new class of cardiovascular drugs, vasopeptidase inhibitors. Omapatrilat is a single molecule that simultaneously inhibits neutral endopeptidase and angiotensin-converting enzyme, thus preserving vasodilator peptides and inhibiting production of the vasoconstrictor angiotensin II. In healthy male volunteers, omapatrilat decreased blood pressure while being generally well tolerated, with no serious adverse events. This study was undertaken to determine the effect of age and gender on the pharmacodynamics of omapatrilat. Healthy male or female volunteers between the ages of 18 and 80 were given a single oral dose of omapatrilat 40 mg. Neither age nor gender affected the vasopeptidase inhibition by omapatrilat. There were no differences between subject groups in the effect of omapatrilat on supine systolic, diastolic, or mean arterial blood pressure. Based on this study of healthy subjects, it can be concluded that it is not necessary to adjust the initial dose of omapatrilat for the treatment of hypertension based solely on age or gender. PMID- 11413937 TI - Tricuspid valve fibroelastoma in an elderly patient: transesophageal echocardiographic diagnosis and differentiation from a myxoma. PMID- 11413938 TI - Well concealed atrial tachycardia with Wenckebach (Mobitz I) atrioventricular block: digitalis toxicity. PMID- 11413939 TI - Angiotensin II receptor (AT1) antagonists in heart failure after Val-HeFT--Quo Vadis? PMID- 11413940 TI - The devil is in the details. PMID- 11413941 TI - Ethical issues in the management of geriatric cardiac patients. PMID- 11413942 TI - The college health opportunity. PMID- 11413943 TI - Men's reproductive and sexual health. AB - A broad definition of men's reproductive and sexual health (MRSH) includes medical (pathophysiological) matters such as sexually transmitted infections (STIs), developmental anomalies, malignancy, trauma, and infertility. It also includes psychosocial concerns: sexuality, contraception, disease prophylaxis, developmental and lifecycle issues, tobacco and drug use, sexual identity and orientation, and partnership issues. College men, of whom a large majority are sexually active, have a range of MRSH needs, including some that are particular to their age and social environment. To reach men effectively requires approaches that are somewhat different from those used with women. Clinicians in college health services are in an excellent position to help young men recognize the importance of reproductive health and sexual responsibility. College health services therefore should offer men screening; clinical diagnosis and treatment for MRSH conditions; and information, education, and counseling services, in a manner designed to meet their unique needs. PMID- 11413944 TI - Ukrainian medical students' experiences, attitudes, and knowledge about reproductive health. AB - In a 1999 classroom survey of sexual behavior among 689 1st-year medical students at Donetsk State Medical University, Ukraine, 59% of the women and 83% of the men revealed they had "ever" had sexual intercourse. The mean age of first intercourse was 15.7 years for the men and 16.6 years for the women. Thirty-two percent of the students reported they did not use contraceptives at their first intercourse, and 19% said they used no contraceptives at their most recent coitus. Condoms were the most frequent means of contraception, followed by coitus interruptus. Less than 5% used oral contraceptives, and 73% of students reported being afraid to use them. About 6% of the students reported they had had sexually transmitted infections (STI) and 10% had had an abortion. Improving knowledge concerning unwanted pregnancies and STI among Ukraine's future health providers may contribute to improving reproductive health issues in general in Ukraine. PMID- 11413945 TI - Medical complications of female genital mutilation. AB - More than 130 million women are subjected to genital mutilation. Despite increasing efforts to reduce the practice, there are many obstacles to eliminating this 2,000-year-old practice, which is based on strong cultural traditions. As college health clinicians provide care to more international students from countries where female genital mutilation is performed, increased awareness and knowledge of the procedure will enable clinicians to understand and manage its complications. We report a case of obstructive uropathy resulting in hydronephrosis secondary to female genital mutilation and review the medical literature regarding this and other complications of genital mutilation "surgery." PMID- 11413946 TI - Masculinity and men's mental health. PMID- 11413947 TI - Preventing depression in high-risk college women: a report of an 18-month follow up. AB - The authors tested the long-term effectiveness of a cognitive-behavioral group intervention in reducing depressive symptoms, decreasing negative thinking, and enhancing self-esteem in 92 college women aged 18 to 24 years who ere at risk for clinical depression. The women were randomly assigned to either an experimental or a no-treatment control group. The experimental group participated in a 6-week cognitive-behavioral intervention that targeted identification and reduction of negative thinking, using such techniques as thought stopping and affirmations. Data on depressive symptoms, self-esteem, and negative thinking were collected before the intervention and at intervals of 1, 6, and 18 months postintervention. The women in the intervention group experienced a greater decrease in depressive symptoms and negative thinking and a greater increase in self-esteem than those in the control group. The beneficial effects continued over an 18-month follow-up period. These findings support the importance of thought stopping and affirmations as prevention interventions with at-risk college women. PMID- 11413948 TI - Ethnic differences in eating disorder symptoms among college students: the confounding role of body mass index. AB - Eating disorders are among the most common psychopathologies on college campuses. Research on ethnic differences in eating disorder symptoms and prevalence has resulted in conflicting conclusions. Some studies find that particular ethnic groups have a higher prevalence of a symptom; others find that members of that ethnic group have a lower prevalence of the same symptom. The authors explored the role of body mass index (BMI), one potential confound. They used a reliable measure of eating disorder symptoms to assess differences between Hispanic, Asian, and non-Hispanic White college women from two separate samples. After controlling for BMI, ethnic differences in eating disorder symptoms of concern about weight and shape disappeared, but differences in restrained eating remained. Inconsistent findings in the ethnic-difference literature on eating disorders may result from systematic group differences in BMI. Implications for college health programs, counseling, and case finding are discussed. PMID- 11413949 TI - Campaigns for conformity. PMID- 11413950 TI - [DRG--an end or a beginning for German hospitals?]. PMID- 11413951 TI - [Introduction of the new DRG-based reimbursement system in German hospitals--a difficult operation? Experiences and possible solutions from the viewpoint of trauma surgery]. AB - Until recently the costs of patients in German hospitals have mainly been calculated according to the length of hospital stay. However, in December 1999, a dramatic change was announced in the social laws in the "Gesundheitsreform 2000." Beginning in 2003 a prospective payment system based on the classification of the "Australian Refined DRGs (AR-DRG)" will be introduced. DRGs are already used in quite a lot of industrialized countries and basically are "per case" payment systems that group patients with homogeneous average costs based on the diagnoses and procedures performed in the hospital. When preparing for this new system, the clinician has a lot of additional tasks. Besides correctly documenting all clinical findings with the ICD-10 Diagnoses and the German OPS301 procedure codes, a knowledge of the economical impact of the clinical decision is absolutely crucial. The most important task is the optimizing of all clinical treatment processes (e.g. by the introduction of clinical pathways), because only hospitals that can do highly efficient treatment will be able to survive in the upcoming competitive situation. In the Krankenhaus Munchen--Schwabing DRGs have been used as a benchmarking tool since 1997. Based on valuable experience, many direct measures to optimize efficiency have been taken. Especially in patients with multiple trauma, it became evident that the use of efficient and standardized treatment can bring economic gain without loss of quality. PMID- 11413952 TI - [DRG--DRG--DRG without end]. PMID- 11413953 TI - [Acute compartment syndrome. Results of a clinico-experimental study of pressure and time limits for emergency fasciotomy]. AB - BACKGROUND: Acute compartment syndrome of the leg is to be regarded as a traumatological emergency. Most specialists already agree that only a timely operative decompression of the afflicted compartment can prevent serious tissue damage. What still remains subject to discussion, however, is the precise tissue pressure above which the operation becomes imperative. Experimental human studies focusing on tissue pressure and muscle oxygenation have not yet been carried out. It was thus the aim of the present study to analyze oxygen partial pressure of the anterior tibial muscle and peroneal action potential in a model compartment syndrome in man. METHODS: In 22 healthy, normotensive volunteers, constant pressure values from 0 to 100 mmHg were induced in the anterior tibial muscle with antishock trousers. Over a period of up to 6 h measurements were made of (1) tissue pressure, (2) intramuscular oxygen partial pressure (pO2), and (3) muscle response potential (MRP) of the n. peroneus profundus by electroneurography. RESULTS: We achieved a 97.7% (Q25%/Q75%: 89.2/99.8) transfer of the pneumatic pressure to the lower leg. Already at intramuscular tissue pressures of 30-40 mmHg, hypoxia and reduction of MRP appeared. A reduction of the MRP to zero and pO2 < 1 mmHg was observed from a pressure of 50 mmHg. Tissue pressure values of over 75 mmHg resulted almost without exception in anoxia of the muscle. CONCLUSIONS: Even under normal perfusion conditions, already slight increases in pressure of above 30 mmHg lead to reduced tissue oxygenation and neural function. We have to consider that with additionally traumatized muscle the ischemic tolerance is markedly reduced and due to unknown influences such as local vasoreactivity and capacity of autoregulation the nutritive perfusion cannot be determined. In the case of a severely injured muscle, to be on the safe side decompressive fasciotomy should therefore be carried out if pressure values remain above 30 mmHg. PMID- 11413954 TI - [Effect of various suture strength factors on behavior of meniscus sutures in cyclic loading conditions]. AB - The aim of this study was to analyze meniscal sutures under cyclic loading conditions for different suture types (vertical and horizontal mattress sutures) and suture materials (absorbable monofilament sutures: PDS 2-0; PDS-0, and PDS-1 USP). Testing was performed on medial porcine menisci, using a well-established biomechanical testing model with a complete longitudinal tear 3 mm from the periphery of the meniscus. Sixty specimens were used. One suture was tested at a time. During cyclic testing 100 load cycles were applied with a crosshead speed of 50 mm/min. Three different maximum loads (10 N, 20 N, and 40 N) were used. The preload was set at 5 N. After cyclic loading, the specimens were loaded until failure. During cyclic loading, a gap appeared between the two parts of the meniscus, and partial tissue failures were observed at the surface of the meniscus. Gapping was more marked with higher loads and with the weaker suture material (p < 0.001). Using PDS 0 and PDS 1 sutures, less partial tissue failures were observed compared to PDS 2-0 (p < 0.001). The ultimate failure loads after cyclic loading were higher with PDS-0 and PDS-1 sutures. With these suture materials vertical sutures were stronger than horizontal sutures (p < 0.05). Using PDS 2-0 this difference could not be found. These results show that the primary strength of meniscal sutures depends on the suture material. The frequency and the amount of gapping and partial tissue failures, which can be observed under cyclic loading, are less distinct with PDS-0 and PDS-1 compared to PDS 2-0. From a biomechanical point of view, PDS 0 and PDS 1 sutures are recommended for meniscal sutures to guarantee a high primary stability, a small amount of gapping, and few partial tissue failures. PMID- 11413955 TI - [Abdominal impalement injuries]. AB - Impalement is an uncommon and spectacular injury, which combines aspects of both blunt and penetrating trauma. With reference to our own seven patients we discuss the initial management and the operative treatment of this rare injury. Further we demonstrate the imminent problems after impalement injuries such as removal of the impaled object, treatment of colon- and other abdominal organ injuries, and management of vascular injuries. PMID- 11413956 TI - [Pronation and supination of the forearm. With special reference to the humero ulnar articulation]. AB - The development of a kinematic model of the pro- and supination, that can be used to predict the influence of angulations of ulna and radius on the pronation and supination is based on the precise knowledge of the pronation and supination movement. We performed two parallel studies for examining the pronation and supination motion of the human forearm. The first experiment dealt with MRI studies on 18 probands (36 examined forearms). As a result we observed an evasive movement of the ulna during the rotation of 7, 14 degrees medial. In order to prove whether the evasive movement was caused by a rotation of the humerus or by an evasion in the articulatio humeroulnaris, we carried out a second experiment, using 30 preparations. The measurement of the pro- and supination motion with a fixed humerus was expedited using a special experimental setup which guaranteed that the ulna could move freely. In all cases we found the same magnitude of the evasive motion of the ulna. Therefore we demonstrated, that the ulna performs an evasive motion during the pro- and supination motion of the forearm that influences the kinematic behavior of the pro- and supination motion significantly. PMID- 11413957 TI - [Supracondylar femoral fractures in knee endoprostheses. Stabilizing with retrograde interlocking nail]. AB - The treatment of supracondylar fractures of the femur in total knee arthroplasty is still challenging, and a variety of methods has been recommended. In a retrospective analysis, we reviewed six patients (average age: 70.5 years) with this type of fracture that had been stabilized with a retrograde intramedullary locking nail [Green-Seligson-Henry (GSH) nail]. The fracture had occurred 34.5 months after implantation of total knee arthroplasty. The average time of the operation was 97.16 min. There were no intra- or postoperative complications. All patients could be followed up at 17.3 months on average. Fracture healing was uneventful in all six cases. The postoperative range of motion was similar to the prefracture level in five patients. One patient demonstrated a loss of extension (10 degrees) associated with a valgus malalignment of 10 degrees. According to our experience, retrograde intramedullary nailing of supracondylar fractures in total knee arthroplasty is a promising alternative, which allows closed reduction and preservation of the soft tissue envelope. Immediate mobilization with partial weight bearing is possible, and the rate of complications is low. PMID- 11413958 TI - [Knee joint prosthesis implantation after fractures of the head of the tibia. Intermediate term results of a cohort analysis]. AB - A significant number of patients with operated tibial plateau fractures develop osteoarthritis and require total knee arthroplasty. In cases of primary osteoarthritis the results are generally good. However, it is not known whether patients with post-traumatic osteoarthritis obtain comparably favorable results. In a retrospective study we analyzed 72 patients who had undergone a self aligning (SAL) total knee arthroplasty. Ten patients received arthroplasty due to sequelae of a tibial head fracture (group I). The median time to follow-up in this group was 30 months. Clinical and radiological evaluation was based on the Knee Society Clinical Rating System. The score comprises pain, range of motion, stability, and function. We defined the axis and possible loosening by radiological examination. For comparative descriptive statistics, a cohort of patients was chosen who had received an arthroplasty because of primary gonarthrosis (group II, 76 arthroplasties in 62 patients). The median time to follow-up in this group was 46.5 months. Three patients in group I underwent revision surgery, four patients displayed severe functional deficits and pain, and one patient had a varus deformity with good clinical function. This corresponded to an early complication rate of 27% and a late complication rate of 36%; a relevant instability or loosening of the components did not occur in this group. In group II the incidence of early complications was 10%. The patients in group I reached a mean value of 153 points using the rating system vs 167 points in group II. Analyzing the clinical parameters of the score, we found that patients in group I experienced significantly more pain, thereby affecting functions of daily living, such as walking and climbing stairs. Because of the small number of patients in group I, conclusions can only be drawn to a limited extent. However, we saw that these patients displayed a higher incidence of complications and performed less well. This has to be taken into consideration and discussed prior to surgery. PMID- 11413959 TI - [A new concept for integration of image databanks into a comprehensive patient documentation]. AB - Image processing and archiving are of increasing importance in the practice of modern medicine. Particularly due to the introduction of computer-based investigation methods, physicians are dealing with a wide variety of analogue and digital picture archives. On the other hand, clinical information is stored in various text-based information systems without integration of image components. The link between such traditional medical databases and picture archives is a prerequisite for efficient data management as well as for continuous quality control and medical education. At the Department of Orthopedic Surgery, University of Berne, a software program was developed to create a complete multimedia electronic patient record. The client-server system contains all patients' data, questionnaire-based quality control, and a digital picture archive. Different interfaces guarantee the integration into the hospital's data network. This article describes our experiences in the development and introduction of a comprehensive image archiving system at a large orthopedic center. PMID- 11413960 TI - [Intra-articular and plantar pressure distribution of the ankle joint complex in relation to foot position]. AB - Ankle injuries are often followed by degenerative changes in the hindfoot joints. Knowledge about the pressure distribution of the intact ankle joint may help to understand the mechanisms leading to cartilage damage. Therefore, we determined the intraarticular and plantar pressure distribution of the ankle joint complex and the Chopart joints with varying foot positions. 12 human lower leg specimens were axially loaded in a foot-loading simulator with full body weight (600 N). A capacitive pressure distribution platform was used to determine plantar pressure patterns. The intraarticular loading situation was measured with Fuji Prescale film. 3 different foot positions (neutral, 10 degrees dorsiflexion, 10 degrees plantarflexion) were investigated. Dorsiflexion led to an increase of the intraarticular contact area, force and mean pressure in the hindfoot. Plantarflexion instead increased loading in the Chopart joints. In the plantar pressure distribution force and peak pressure under the hindfoot increased with dorsiflexion. With plantarflexion area, force and peak pressure under mid- and forefoot increased. With our study we could demonstrate that the loading situation of the ankle joint complex is significantly influenced by the foot position. These findings may help to understand the development and localisation of arthritic changes due to posttraumatic changes of the joint loading characteristics. PMID- 11413961 TI - [Fractures of the distal humerus in the child]. PMID- 11413962 TI - [Multiple pathological fractures within the scope of DeToni-Debre-Fanconi syndrome after fumarate therapy in psoriasis]. AB - We report about a rare case of a pathological fracture of the shank following earlier pathological fractures at other locations in a comparatively young female patient with no history of trauma. There were no known diseases other than psoriasis. The shank fracture was treated surgically by osteosynthesis. Osteoporosis, myeloma, or malignancy as causative factors of this fracture could be excluded. Scintigraphy showed an enhancement, especially at the extremities. Other than reactive bone growth, histological examination revealed no further aspects. Laboratory analysis indicated a massive lack of vitamin D3. After transferring the patient to the internal department of our hospital, long-term medication with fumaric acid was determined to be the reason for the osteomalacia of a Fanconi's syndrome. Three months after cessation of these medicaments and treatment with active vitamin D3 metabolites, the patient was free of complaints. The radiographs showed an essential improvement of the demineralization. PMID- 11413963 TI - [Parosteal lipoma]. AB - Paraosteal lipoma is a rare benign tumour of the musculoskeletal system. It's features consist of a fatty component seated on a hypersotosis i.e. an exostose like bone prominence. This tumor has a high tendency of metaplasia. Therefore chondroid elements may be present in addition to the fatty and bony components. Therapy of choice is complete surgical removal including the hyperostosis. Presenting this case the clinical and morphological characteristic of this rare tumour are discussed. PMID- 11413964 TI - [Pathological fracture in osteofibrous dysplasia]. AB - Osteofibrous dysplasia Campanacci is a rare tumor-like lesion mostly presented in the meta-epiphyseal region of the tibia. Different options exist about the treatment modalities. The rare case of a 4 1/2-year-old girl suffering from a pathologic fracture of the tibia shaft due to a osteofibrous dysplasia is presented. The fracture was stabilized by intramedullary nailing using elastic titanium rods. During this procedure a biopsy of the lesion confirmed the radiological suspected diagnosis of an osteofibrous dysplasia. The implant was removed after ten months when fracture and lesion were healed. The treatment options and the differential diagnoses are discussed by review of the literature. PMID- 11413965 TI - [Comment on the contribution by H. Diste/AOK-Rhineland, "Quality-assisted resource control from the viewpoint of public health insurance"]. PMID- 11413966 TI - [Comment on the contribution by Th. Bein et al.: "Ventilation in prone position of a 5-year-old child after polytrauma. Effective therapy of persistent atelectasis"]. PMID- 11413967 TI - [The Galilei process from the viewpoint of Emil Wohlwill]. PMID- 11413968 TI - [Did John Paul II keep his promise?]. PMID- 11413969 TI - [The experiment in Galilei's manuscripts--science in action and myths]. PMID- 11413970 TI - [Galileo Galilei and the Accademico Incognito]. PMID- 11413971 TI - [Galilei, the bone of the giant and the wrong Ariost]. PMID- 11413972 TI - [Proportion science and speed of movement]. PMID- 11413973 TI - [Galilei, Leibniz and the scrupulositas of mathematics]. PMID- 11413974 TI - [Once again on the relationship between Galilei and Kepler]. PMID- 11413975 TI - [Dialogo sopra i due massimi sistemi as a classic]. PMID- 11413976 TI - [Falling bodies and the riddle of Domigo de Soto]. PMID- 11413977 TI - [The modern Galilei]. PMID- 11413978 TI - Investigation of antinuclear antibodies in canine atopic dermatitis. AB - Serum samples from 40 atopic dogs and 20 healthy dogs were assayed for antinuclear antibodies (ANA) using a human epithelial cell line (HEp-2) and standard indirect immunofluorescent methods. Samples from the atopic dogs were grouped according to the presence (n = 28) or absence (n = 12) of facial lesions at any moment during the follow-up period. Positive ANA titres were found in 10 of the 40 atopic dogs analysed (25%) whereas samples from the control group were negative. Eight atopic dogs with facial lesions had a positive titre (28.57%) in contrast with atopic dogs without facial lesions where two positive samples (16.67%) were found; however, the differences were statistically not significant. Endpoint titres were low (1/40), only two samples yielded a 1/80 positive titre and one sample had a 1/320 titre, all of them from the facial lesions group. Response to immunotherapy was classified as positive only in four of the 10 ANA positive patients (40%) compared with a response rate of 73% (22 of 30) in the ANA-negative atopic dogs. However, a Fisher's exact test showed a two-sided P value of 0.122 which was considered statistically not significant. The overall response rate to immunotherapy for all atopic dogs was 65% (26 of 40). In conclusion, the prevalence of ANA is higher in atopic than in healthy dogs, especially if facial lesions are present. Although a clinically significant pathogenic contribution is not probable, this higher prevalence should be taken into account in the differential diagnosis of canine autoimmune dermatitis. PMID- 11413979 TI - Possibilities of flow cytometric analysis for immunophenotypic characterization of canine lymphoma. AB - The purpose of the study presented here is to show the possibilities of flow cytometric analysis for immunophenotypic characterization of canine malignant lymphoma. Aspiration biopsies from lymph nodes of 30 dogs with multicentric lymphoma were stained by four dog-specific antibodies and three human-specific antibodies cross-reacting with canine lymphocytes, and analysed by flow cytometry. Twenty of the 30 lymphoma examined were characterized as B-cell lymphoma and 10 as T-cell-lymphoma. Flow cytometry is a suitable method for the immunophenotypic analysis of canine lymphoma. Contrary to immunohistochemistry, no surgically produced lymph node biopsy is required and the results are available within a few hours. PMID- 11413980 TI - Protein peroxidation processes in bovine retained and not-retained placenta. AB - Proteins as well as lipids and other biologically active substances may undergo peroxidative damage caused by reactive oxygen species (ROS). The aim of this study was the determination of this damage in cases of retained (RFM) and not retained bovine foetal membranes. The levels of thiol groups were measured spectrophotometrically. Tryptophan, formylokinurenine and bityrosine bridges were determined using spectrofluorimetric methods. All parameters were determined in placental maternal and foetal homogenates collected from cows divided into six groups as follows: A, pre-term caesarian section without RFM; B, pre-term caesarian section with RFM; C, term caesarian section without RFM; D, term caesarian section with RFM; E, spontaneous delivery without RFM; F, spontaneous delivery with RFM. All parameters except tryptophan showed statistically significant differences between the maternal and foetal parts of the placenta. The concentrations of the thiol groups were lower in RFM than in not-RFM cases, whereas formylokinurenine and bityrosine concentrations showed the opposite relationship. Tryptophan levels were lower in cases of RFM rather than not-RFM samples but statistically significant differences occurred in groups C and D and the foetal part of groups E and F. Further experiments are necessary to determine the oxidative status of other biologically active substances in cases of retained bovine placenta. PMID- 11413981 TI - The application of an indenter system to measure structural properties of articular cartilage in the horse. Suitability of the instrument and correlation with biochemical data. AB - A recently developed indenter system that aims at determination of local structural properties of the cartilage surface was evaluated for suitability in the horse. To this aim, maximum indenter force was measured of the articular surface and related to biochemical characteristics of the cartilage at different sites of the distal metacarpal bone (MC). Significant topographical variation exists in structural properties of the articular surface of the MC. The dorsal margin showed a significantly lower maximum indenter force than more centrally located areas, indicating an increased compliance under load. A high maximum indenter force correlated positively with high glycosaminoglycan levels. No correlation was found between measured indenter force and collagen content of the cartilage. It was concluded that the instrument is able to quantify differences in structural properties of the cartilage surface which reflect glycosaminoglycan content and hence is capable of giving some indication of cartilage quality. The instrument is a potentially useful tool, but use is limited due to geometrical constraints. PMID- 11413982 TI - Comparison of sevoflurane with isoflurane for rapid mask induction in midazolam and butorphanol-sedated dogs. AB - Rapid mask induction can be a useful induction technique for veterinary patients, although it is often accompanied by exaggerated excitement responses in unpremedicated animals (Mutoh et al.: Jpn. J. Vet. Anesth. Surg. 26, 109-116; J. Vet. Med. Sci. 57, 1007-1013; J. Vet. Med. Sci. 57, 1121-1124; 1995). The aim of this study was to compare sevoflurane with isoflurane for rapid mask induction in six dogs sedated by a combination of midazolam (0.1 mg/kg) and butorphanol (0.2 mg/kg). Induction with sevoflurane (5%, 2.4 minimum alveolar concentration [MAC]) in O2 resulted in shorter time to loss of the palpebral reflex, negative tail clamp response, and successful intubation than with isoflurane (3%, 2.4 MAC) in O2. There were no changes in heart rate or mean arterial blood pressure during induction with sevoflurane, whereas an increase in heart rate was observed in dogs induced with isoflurane. A decrease in respiratory rate compared with the pre-induction rate was observed during induction, and associated mild respiratory acidosis, characterized by an increase in arterial PCO2, was measured at the end of the induction period in both induction groups. None of the animals had episodes of induction-related complications. These results suggest that both sevoflurane and isoflurane produce a smooth onset of induction in midazolam and butorphanol-sedated dogs. Sevoflurane is a more suitable for rapid mask induction than isoflurane since it provides faster induction associated with a lower blood/gas partition coefficient. PMID- 11413983 TI - Comparison of medetomidine and dexmedetomidine as premedication in isoflurane anaesthesia for orthopaedic surgery in domestic sheep. AB - The objective of the present study was to determine the potency of dexmedetomidine in relation to medetomidine in sheep undergoing orthopaedic surgery by comparing the anaesthetic requirements and cardiovascular changes at a dose relationship that represented equipotency in vitro. Twenty-four non pregnant, female sheep were used. The study was carried out as a blind, randomized, experimental trial. Group 1 received 5 micrograms/kg bodyweight (BW) dexmedetomidine and group 2 received 10 micrograms/kg BW medetomidine intravenously 5 min prior to induction of anaesthesia. Anaesthesia was induced with ketamine (2.0 mg/kg BW intravenously) and maintained with isoflurane in 100% oxygen. End expired anaesthetic concentration (FEIso), end expired carbon dioxide concentration (FECO2), respiratory frequency (fR), direct arterial blood pressures, heart rates (HR) and arterial blood gases were monitored. Data were averaged over time and tested for differences between groups by independent t tests, and analysis of variance for repeated measures. Average FEIso concentrations required to maintain a surgical plane of anaesthesia were not different between groups (1: 1.02 +/- 0.04%; 2: 0.99 +/- 0.07%). There was no difference in HR, arterial blood pressures, fR, FECO2 and arterial blood gases between groups. Average mean PaO2 were 279.54 +/- 113.37 mmHg and 220.21 +/- 102.15 mmHg with individual minimum values of 27.2 mmHg and 58.5 mmHg in groups 1 and 2, respectively. In conclusion, intravenous dexmedetomidine at 5 micrograms/kg BW and medetomidine at 10 micrograms/kg BW have the same effects on isoflurane requirements and cardiopulmonary parameters in sheep, indicating an equipotent dose relationship. Both preparations induced moderate to severe hypoxaemia in individual sheep. PMID- 11413984 TI - Liver and biliary system pathology in equine dysautonomia (grass sickness). AB - Hepatocellular and hepatobiliary damage was assessed in equine acute, subacute and chronic grass sickness cases (AGS, SAGS, CGS). Histopathological analysis showed that even in some early AGS cases enlarged hepatocytes, hepatocyte vacuolation indicative of lipid accumulation (steatosis), intrahepatocyte, canalicular and periportal deposition of pigments, frequent leucocyte infiltration and cholangitis occurred. Analysis of serum indicated significantly increased levels of unconjugated bilirubin in all groups and conjugated bilirubin in AGS and SAGS groups, increased levels of bile acids in some individuals from each group and significantly increased levels of glutamate dehydrogenase (GLDH) in AGS and SAGS cases. Conjugated bilirubin was significantly elevated in urine of AGS and SAGS cases. The evidence suggests that abnormal liver function involving moderate hepatocellular pathology in conjunction with steatosis and cholestasis may contribute to the pathogenesis of GS. PMID- 11413985 TI - [The references in articles published in biomedical journals]. AB - Bibliographic citations or "references" are an important component of all scientific manuscripts. The authors are responsible for their accuracy and they should follow the format and style requested by the journal where they are submitting their paper. Revista Medica de Chile adheres to the "Uniform Requirements" established by the Vancouver Group of Medical Editors. Equally important is a correct choice of references, including those original articles strictly connected to the content of the manuscript. The number of citations usually vary according to the specific character of the study: research article, or case reports, review article, etc. Common mistakes are due to an excessive number of repetitive or irrelevant citations, or the omission of important articles sometimes not found in bibliographic indexes, or an erroneous claim of priority in reporting an observation. Finally, in developing countries the authors should include previous reports appeared in their own local journals, therefore improving their opportunities of achieving international visibility. PMID- 11413986 TI - [Study of cytokines kinetics in severe sepsis and its relationship with mortality and score of organic dysfunction]. AB - BACKGROUND: The Infectious Systemic Inflammatory Response syndrome and multiple organic dysfunction have common physiopathological mechanisms. Multiple organic dysfunction can be assessed using severity scores. AIM: To relate cytokine kinetics with a multiple organic dysfunction score during sepsis. MATERIAL AND METHODS: Tumor necrosis factor alpha (TNF alpha) and interleukin 6 (IL6) kinetics were studied in 25 patients with severe sepsis with less than 48 h of evolution and interleukin 1 beta (IL beta) kinetics was studied in 13 patients. Measurements were made at 0, 12, 24 and 48 hours after admission to the study, using an ELISA technique. These parameters were correlated with the Marshall multiple organic dysfunction score and survival. RESULTS: Mean age of study subjects was 70 years, the APACHE II score was 16.9 +/- 6 and the Marshall score was 6.8 +/- 3.6. Sepsis was of pulmonary origin in 56% of patients and intra abdominal in 32%. Mortality was 36%. TNF alpha increased during the study period (24.1 pg/ml initially and 37.8 pg/ml at 24 hours, with a slight posterior reduction, p < 0.02). These levels had no association with mortality or organic dysfunction. IL6 remained elevated during the first hours and had a tendency to decrease thereafter. Decreased patients had higher values than survivors (306 pg/ml and 55.4 pg/ml respectively, p = 0.011). Its values were tightly correlated with Marshall score, with the number of failing organs, with the presence of shock and with probability of dying during hospitalization. IL1 beta remained low and was not associated with clinical parameters. CONCLUSIONS: There is a tight correlation between the elevation of IL6 and the severity of the Systemic Inflammatory Response and mortality in these patients with sepsis. PMID- 11413987 TI - [Quality of life in patients with chronic obstructive pulmonary disease and the impact of physical training]. AB - BACKGROUND: Health related quality of life (QoL) is severely impaired in COPD patients as a consequence of dyspnea and limited exercise tolerance, which lead to physical deconditioning and muscle atrophy resulting in weakness and fatigue. Psychosocial factors such as depression and anxiety also contribute to this impairment. AIM: To evaluate: a) the impact of COPD on quality of life, and b) the effect of 10 weeks of exercise training on exercise performance and on QoL. PATIENTS AND METHODS: The Spanish version of the Chronic Respiratory Questionnaire (CRQ) was applied to 55 COPD patients (FEV1 37 +/- 13% pred) for the assessment of QoL and in 30 of them submitted to exercise training for 10 weeks. Exercise performance was evaluated by measuring: six-minute walking distance, maximal workload (Wmax), maximal O2 consumption (VO2max) as well as endurance time, blood lactic acid, dyspnea and leg fatigue during a submaximal exercise. Trained patients were evaluated before and after training. RESULTS: COPD patients showed a reduction (mean +/- SD) in the four domains of the CRQ: dyspnea (3.1 +/- 0.9); fatigue (4.3 +/- 1.3); mastery (4.65 +/- 1.3), emotional function (4.1 +/- 0.97), and in Wmax and VO2max (52 +/- 16 Watt and 970 +/- 301 ml/min). No significant relationship between the impairment in exercise tolerance and in QoL was observed. Exercise training significantly improved the four domains of QoL (p < 0.0001), Wmax (p < 0.05), VO2max (p < 0.02) and endurance time (p < 0.001). Isotime exercise measurements of dyspnea, leg fatigue and lactic acid decreased after training (p < 0.001, each). No significant relation between changes in QoL and changes in exercise performance were observed. CONCLUSIONS: Our results demonstrate that QoL is seriously impaired in patients with COPD and confirm: (a) the lack of relationship of QoL to the usually measured physiological parameters, and (b) the beneficial effect of exercise training on QoL through the reduction of symptoms. These findings stresses the need of measuring quality of life in our patients if we want to evaluate the impact of therapeutic procedures on well-being from the patients' perspective. PMID- 11413988 TI - [Molecular diagnosis of Prader-Willi and Angelman syndromes: methylation, cytogenetics and FISH analysis]. AB - BACKGROUND: The diagnosis of Prader-Willi and Angelman syndromes is difficult, since their phenotypic manifestations are variable and unspecific. The study of the methylation state of DNA in 15(q11-q13) using polymerase chain reaction, called methylation test, allows the diagnosis of most patients with Prader-Willi and Angelman syndromes, irrespective if the underlying molecular alteration is a deletion, uniparental disomy or a punctual imprinting mutation. AIM: To assess the effectiveness of methylation test in the diagnosis of Prader-Willi and Angelman syndromes. PATIENTS AND METHODS: Thirty seven cases with a presumptive diagnosis of Prader-Willi syndrome and 25 with the presumptive diagnosis of Angelman syndrome were studied. Methylation test was done in genomic DNA obtained from peripheral lymphocytes. RESULTS: Methylation test confirmed the clinical diagnosis in 11 of 37 patients with Prader Willi (30%) and 6 of 25 patients with Angelman syndrome (24%). CONCLUSIONS: Clinical criteria overestimate the diagnosis of Prader-Willi and Angelman syndromes. The initial diagnosis should be confirmed with the methylation test and, if necessary, with FISH that will detect most deletions in the region. PMID- 11413989 TI - [Carbohydrate deficient transferrin, gammaglutamyl transferase and mean corpuscular volume in the evaluation of recent alcohol intake in excessive drinkers]. AB - BACKGROUND: There are no reliable markers to detect heavy drinking or as a tool to control abstinence compliance in alcoholic treatments. The Mean Corpuscular Volume (MCV), and the gammaglutamyl transpeptidase (GGT), are widely used although their predictive value is somewhat limited due to their low specificity. On the other hand, the Carbohydrate-deficient transferrin (CDT) described in the eighties is highly specific and would be of value in early detection of problem drinking. AIM: To compare the sensitivity and specificity of CDT, GGT, and MCV in order to evaluate their single and combined use as markers for detection of heavy drinking behaviour. PATIENTS AND METHODS: CDT, GGT, and MCV values were determined in blood samples from (a) alcoholics (drinking more than 100 9 alcohol/day; n = 47) and (b) healthy volunteers, teetotalers from the Church of Saints of Later Days (n = 34). At the time of sampling alcoholics were presently drinking or had been abstinent for no more than six weeks. ROC curves were used to determine the best cut-off point for each marker. RESULTS: Sensitivity was found to be similar for all three markers. Specificity was found higher for GGT (90.9%) and CDT (91.0%). The combined use of MCV, GGT and CDT, that is, when at least one of the markers is altered, was shown to detect 83% of the patients. No correlation was observed between the markers and the level of alcohol intake. CONCLUSIONS: CDT could be of value as a marker to detect heavy drinking when used with GGT and MCV values combined. CDT is particularly higher in drinking alcoholics and remains significantly high for at least six weeks after they stop drinking. PMID- 11413990 TI - [Reference values of growth hormone binding protein (GHBP) for a normal pediatric population]. AB - Circulating concentrations of the high affinity growth hormone binding protein (GHBP) may be a marker of GH receptor density as well as GH sensitivity. GOAL: To determine values of GHBP for a normal Chilean pediatric population. METHODS: We determined GHBP levels in 73 males and 73 females between 4 to 15.5 years and 4 to 16.8 years respectively, divided in 7 groups according to age and puberal status. RESULTS: The population was normally distributed in weight, height and body mass index (BMI). GHBP activity increased up to Tanner IV in males and Tanner III in females, and decreased slightly thereafter in Tanner V and IV respectively. There was a significant difference between GHBP levels of preschool children and those found in Tanner II to V in both sexes (p < 0.05). In addition, we found a positive correlation between GHBP vs weight, height and BMI (p < 0.001) in males and females. CONCLUSION: The availability of this methodology allows us to establish the normative value of GHBP in our population and provides useful information to interpret GH circulating levels in children with growth disorders. PMID- 11413991 TI - [Digestive refeeding in acute pancreatitis. When and how?]. AB - BACKGROUND: Digestive refeeding in acute pancreatitis represent a difficult issue. It requires the resolution of intestinal ileus and carries a risk of reactivation. AIM: To evaluate criteria that may guide in early refeeding avoiding unnecessary prolonged fasting. PATIENTS AND METHODS: Thirty patients with acute pancreatitis were evaluated in a prospective trial. The severity of the pancreatitis was evaluated according to APACHE II score and Balthazar CT scan alterations. The criteria proposed to start early refeeding were absence of nausea and vomiting, decreased abdominal pain, presence of bowel sounds and lowering of serum amylase levels. Balthazar CT scan classification, was used to decide between oral or enteral refeeding. RESULTS: Eighty percent of patients had alterations in pancreatic density, necrosis or pancreatic or peripancreatic liquid collections in the CT scan (corresponding to Balthazar stages C, D, or E). Ten patients fullfilled the criteria for enteral refeeding at 8.1 +/- 3.5 days (range 3 to 15 days), and 21 patients fulfilled criteria fo enteral refeeding at 8.7 +/- 4.5 (range 4-19). No patient had a reactivation of his pancreatitis. CONCLUSIONS: Digestive refeeding can be done safely by using the criteria proposed in this study. Pancreatic necrosis or peripancreatic fluid collections do not contraindicate refeeding. Oral feeding may be employed (as the first option) in selected patients, without increasing the risk of complication, regardless of CT scan alterations of the pancreas. PMID- 11413992 TI - [Complications in children with varicella in 4 hospitals in Santiago, Chile: clinical spectrum and estimation of direct costs]. AB - BACKGROUND: The knowledge of varicella complications and their associated cost may help for a better evaluation of varicella immunization benefits. AIM: To determine frequency, type, outcome and affected population of varicella complications in children requiring hospitalization, and to estimate their direct costs. MATERIAL AND METHODS: Retrospective analysis of medical records of children admitted to four hospitals in Santiago, Chile, due to varicella complications between January 1997 and February 1999. Calculation of direct costs of hospitalizations in a sample of 30 patients. RESULTS: One hundred fifty four patients were identified, 74% were younger than 5 years old, only one was immunocompromised. Complications identified were skin and soft tissue infections in 63%, invasive infections in 25.3%, neurological in 7.1% and miscellaneous in 4.5%. Staphylococcus aureus and Group A beta-haemolytic Streptococcus (GABS) were predominantly isolated. S. aureus was the main agent identified in superficial infections and GABS in invasive infections (sterile sites). Two patients died due to invasive infections (streptococcal toxic shock and S. aureus septicaemia) and 11 required surgical procedures. The average cost per hospitalization was US$ 600 in public hospitals and US$ 1,800 in the private hospital. CONCLUSIONS: Varicella complications requiring hospitalization are due mainly to bacterial infections and they affect immunocompetent toddlers. These complications can be severe and even fatal. PMID- 11413993 TI - [Infant mortality differentials from congenital malformations with paired data. Chile (1993-1995)]. AB - BACKGROUND: The analysis of infant mortality from congenital malformations, which at present is the main group of causes of this mortality in Chile, suggests that it could be decreased with a good knowledge of its conditioning factors. AIM: To study infant mortality differentials from congenital malformations with linked records, in the 1993 to 1995 Chilean birth cohorts. MATERIAL AND METHODS: Analysis of mortality differentials in 1993, 1994 and 1995 birth cohorts. Multivariate logistic regression of mortality from congenital diseases. RESULTS: Univariate analysis showed that mortality is highest in the Southern regions of the country (VII to XII) and in rural areas. It is also higher in children from older and from very young mothers, it increases along with the birth order of the child and decreases with increasing educational level of the mother. Multiple logistic regression analysis, confirmed the higher mortality in the Southern regions, aged mothers, high birth order of the child and low educational level of the mother. However no significant influence of rurality nor greater mortality in children of very young mothers was found. CONCLUSIONS: These results can be attributed to the fact that this type of analysis permits the control with other variables. Although the mortality data showed interesting relationships with the independent variables, a registry of all live births and stillbirths with congenital anomalies, that would provide greater numbers and data on non fatal anomalies, would be desirable to better study their causal factors. PMID- 11413994 TI - [Assessment of techniques and errors in the use of metered dose inhalers in the adult patient]. AB - BACKGROUND: Not all the techniques for the correct use of metered dose inhalers are used by patients and health care professionals. AIM: To assess the techniques and errors in the use of metered dose inhalers among patients and health care professionals. MATERIAL AND METHODS: Evaluation of the inhaling technique, using a validated questionnaire, used by 68 patients, 30 physicians and 30 nurses working in a hospital at Concepcion, Chile. RESULTS: The "closed mouth" technique is used by 84% of patients, 40% of physicians and 73% of nurses. The rest uses the "open mouth" technique. The aerochamber is used by 12% of patients, 37% of physicians and 27% of nurses. The most frequent inhaling errors detected were not shaking the inhaler (26% of patients, 30% of physicians and 7% of nurses), not applying the puff at the start of inspiration (28% of patients, 7% of physicians and 13% of nurses), and not maintaining an apnea after the inhalation (41% of patients, 7% of physicians and 10% of nurses). Sixty percent of patients, 67% of physicians and 40% of nurses have not received instructions about the inhaler use. CONCLUSIONS: There is a high frequency of errors in the use of inhalers and most users have not been trained in its use. PMID- 11413995 TI - [Endovascular therapy of superior vena cava syndrome: clinical case]. AB - The treatment of superior vena cava syndrome is usually oriented to the underlying cause, that can be too slow in emergency cases. We report a 49 years old woman with a multiple myeloma that was admitted due to a superior vena cava syndrome caused by a central venous catheter used for chemotherapy for 20 weeks. She was successfully treated with thrombolysis, angioplasty and stent placement. The patient died 7 months later due to the underlying disease. Long term catheters are the responsible for 20 to 30% of superior vena cava syndromes. Endovascular treatment of the syndrome is successful in 60 to 100% of cases with a symptomatic relapse in 4 to 45% of patients. PMID- 11413996 TI - [Central pontine myelinolysis and hyponatremia. Clinical case]. AB - There is a controversy whether central pontine myelinolysis can complicate either hyponatremia or its rapid correction. We report a 69 years old woman with a history of one month of vertigo, nausea, vomiting and diarrhea. She was admitted dehydrated ad stuporous, and initial laboratory values showed a serum sodium of 96 mEq/L. She was treated with dextrose 5% and 3% NaCl. Serum sodium raised to 120 mEq/L at the next day and the level of consciousness improved. At the 4th day of admission, the patient was again stuporous and with spastic quadriplegia. A magnetic resonance imaging showed a central and symmetrical pontine lesion on T1 and T2 weighed images. Thereafter, the patient experienced a progressive improvement of her neurological condition and was discharged three months later, moving her lower limbs. Nine month later she was able to walk. PMID- 11413997 TI - [Are transdermic estrogens cardioprotective?]. AB - Transdermic estrogens share many of the oral estrogens cardiovascular effects, but so far there are no studies proving that they have a cardioprotective effect neither in animals nor in human beings. The doubt is outlined moreover, when most of the investigations performed with oral estrogens in animals show an antiatherogenic effect, while the few experimental studies that hare been carried out with estrogen patches show contradictory results. We will have to wait for more extensive clinical trials to be able to know if the transdermic estrogens are really cardioprotective, however if we want to achieve some cardiovascular risk improvement with the current knowledge we will probably have to support the use of oral estrogens. PMID- 11413998 TI - [Ontogenetic origin of the human individual from his genome]. AB - Diverse propositions about the ontogenetic origin of a live organism, specially human beings, are examined. Unambiguous and objective propositions about this origin are that a live organism is an ontogenetically programmed and integrated organisation, that the origin condition has the greater influence on other processes, that in pluricellular organisms, no organ or tissue can be considered critical to establish origins and that the origin must be established by endogenous elements. Several hypotheses about the origin of life are discarded. The integration between oocyte cytoplasm and the genetic material that it receives, that culminates in the first genome replication, is proposed as the process that gives origin to the individual. This process occurs in all living organisms. PMID- 11413999 TI - [History of the scientific work of Eduardo Cruz-Coke Lassabe]. AB - BACKGROUND: Eduardo Cruz Coke M.D., (1899-1974) was one of the precursors and pioneers of biomedical research in Chile, as professor of Physiological and Pathological Chemistry at the University of Chile, from 1925 to 1962. He was a disciple of Dr. Juan Noe and studied in Europe with the Nobel Prize winners Otto Warburg, Jean Perrin, Louis de Broglie and Frederic G. Hopkins. In Chile, he founded a scientific academy with disciples that later obtained the National Sciences Award, such as Hector Croxatto, Jorge Mardones, Hermann Niemeyer, Luis Vargas and Jorge Allende. He carried out pioneering research in metabolism, nutrition, endocrinology, oncology and nephrogenic hypertension. He published more than 50 scientific papers in French, English and Spanish. He founded scientific societies, edited journals and created the National Commission of Nuclear Energy. His books were "The ionic acidity in the clinic", "Preventive and directed medicine", "The adrenal cortex". He was Ministry of Health between 1937 and 1938 and passed important socio-medical bills. He obtained the distinguished international awards in Europe, the U.S.A. and Latin America. The Biomedical Sciences Institute of the University of Chile carries his name. PMID- 11414000 TI - [Professor Dr. Abraham Horwitz Barak (1910-2000)]. AB - Abraham Horwitz, MD, was a prominent leader in Public Health in the Americans. His career started as a clinician and researcher in infectious diseases, then as a teacher in two Chilean Universities, one of the organizers of the Public Health Service in Chile and, subsequently, a prominent staff member in the Pan American Health Organization, becoming its Director during four consecutive terms. His outstanding career was recognized worldwide with several honorary awards and distinctions. Dr. Horwitz is recognized as one of the most distinguished and influential individuals in Medicine, in the American Continent, during the 20th Century. PMID- 11414001 TI - [Cumulative ionizing radiation in the medical radiodiagnosis]. PMID- 11414002 TI - [Methodological errors in mainstream journals]. PMID- 11414003 TI - Focused attention reduces the effect of lateral interference in multi-element arrays. AB - Lateral interference of nearby irrelevant flankers may be reduced when attention is already focused at a relevant position. This hypothesis was tested in three experiments with multi-element arrays consisting of one target and several neutral flankers with reaction time (RT) and proportion of errors (PE) as dependent variables. The arrays were preceded by peripheral cues that varied in size to induce different levels of focused attention. In a first experiment, the eccentricity effect was shown to be affected by attention, but no support was found for a reduction of lateral interference in the case of focused attention. However, these results may be due to an attentional bias for central elements. Two additional experiments were performed in which target-flanker distance was manipulated. RTs and PEs showed that the effect of target-flanker distance was smaller when the precise target position was indicated by a peripheral precue. Thus, focused attention seems to reduce the effect of lateral interference. These results can be explained most easily by a model in which attention already affects early perceptual processing. PMID- 11414004 TI - Frames of reference in perceptual-motor learning: evidence from a blind manual positioning task. AB - Participants moved a joystick to bring a computer-displayed cursor to each of six on-screen target locations arrayed around the center of the screen. At the start of each trial, the stick rested vertically, with a cursor occupying the center of the screen. A target appeared at another location and as soon as the stick was moved away from its rest position the cursor disappeared until the participant pressed a trigger on the stick to indicate when s/he thought the stick-controlled cursor was at the target site. With training, participants improved on the blind positioning task, but when conditions changed their performance suffered. Changing the hand used in the task or the location of the stick caused approximately equal disruptions, but changing both hand and location was significantly more disruptive than changing just one feature. The results support the hypothesis that perceptual-motor learning entails coding of extrinsic (spatial coordinates) as well as intrinsic (postural or body movement) information. PMID- 11414005 TI - Misinterpretation of conditional statements in Wason's selection task. AB - Errors may be made on Wason's selection task because either (a) the rule to be tested is misunderstood, or (b) reasoning from that rule is inaccurate, or both. We report two experiments using the experimental paradigm introduced by Gebauer and Laming in which subjects are given six problems in succession. We use the subset of cards selected by each subject as (a) an indication of how the rule is understood and, when that selection is consistent throughout all six problems (so that we can infer a consistent understanding of the rule), as (b) a basis for evaluating the accuracy of the subject's reasoning according to three independent criteria. Experiment 1 adds an exactly parallel contextual version of the task to permit comparison between performances (by the same subjects) on the two versions. Experiment 2 repeats Exp. 1, but with negatives inserted in the conditional rule. Most subjects make a consistent selection of cards throughout all six problems, but typically appear to misunderstand the rule. This is so in both abstract and contextual tasks and replicates the finding by Gebauer and Laming. Most misunderstandings consisted of either (a) reading the simple conditional rule as a bi-conditional or (b) substituting "top/underneath" for "one side/other side". In Exp. 1 subjects seldom misevaluated the rule they appeared to be testing, but such "errors" of evaluation were common in Exp. 2. Negatives confuse the subjects and should not be used in any conditional application that matters. In Exp. 2 (but not 1) there was a significant correlation between interpretations of the two tasks. We provide an explanation of "matching bias" (it results from the confluence of the two common misunderstandings above) and comment on "mental models" which are, at present, unable to accommodate the variety of results we present here. We also relate our experimental paradigm to the conditional inference task and to truth tables. PMID- 11414006 TI - Simultaneous learning of different regularities in sequence learning tasks: limits and characteristics. AB - Two experiments are reported which were designed to investigate the generality and the power of the mechanisms underlying sequence learning. In both experiments, participants reacted to systematic sequences of tones. They were informed that there was a tone systematicity. Participants were not told that the interval between a response to a tone and the onset of the subsequent tone (response-signal interval, RSI) also varied according to a fixed regularity. Experiment 1 showed that the unattended RSIs were learned when they were uniquely related to the tone sequence, but not when the relation was ambiguous. Experiment 2 showed that, on the basis of the traditional reaction time performance measure, participants who learned the RSIs by attending to their systematicity could not be distinguished from those in an incidental learning condition in which the RSI systematicity was unattended. However, a model-based analysis of the processes contributing to judgements about the event sequences suggested that the two groups had acquired qualitatively different knowledge. PMID- 11414007 TI - Global/local processing and negative priming: the influence of selection difficulty and stimulus exposure. AB - Negative priming is a decrement in performance observed when a previously ignored stimulus is re-presented as a target. The present study examined the relation between selection difficulty and negative priming in five experiments that used hierarchical stimuli (large letters made up by small letters). The results show that negative priming is greater when subjects direct attention to the local level (more difficult selection) than when they direct attention to the global level (less difficult selection). However, that occurs only when exposure of prime and probe is sufficiently long. With shorter presentations, negative priming is still observed but is no longer modulated by selection difficulty. These results suggest that both anticipatory and reactive mechanisms are responsible for the occurrence of negative priming and that instantiation of the reactive mechanism depends on the time available for prime and probe selection. PMID- 11414008 TI - Focusing attention on overlapping and non-overlapping figures with subjective contours. AB - In a series of five experiments we investigated whether observers could focus attention on a restricted visual area that was demarcated by Kanizsa-like subjective contours, and whether this effect also occurred in the case of overlapping figures. The task was a simple reaction time to a luminance increment and the basic finding was that reaction time was faster when the imperative stimulus fell inside the focus of attention than when it fell outside. The first two experiments showed that the extent of the attentional focus could be adapted to a region that was demarcated by subjective contours, thus extending the results of previous studies that used regions demarcated by real contours. The last three experiments showed that, regardless of the type of margins, focusing was more efficient for the figure that was perceived as lying in front in a pair of overlapping figures. PMID- 11414009 TI - Estimated effects of temperature on secondary organic aerosol concentrations. AB - The temperature-dependence of secondary organic aerosol (SOA) concentrations is explored using an absorptive-partitioning model under a variety of simplified atmospheric conditions. Experimentally determined partitioning parameters for high yield aromatics are used. Variation of vapor pressures with temperature is assumed to be the main source of temperature effects. Known semivolatile products are used to define a modeling range of vaporization enthalpy of 10-25 kcal/mol-1. The effect of diurnal temperature variations on model predictions for various assumed vaporization enthalpies, precursor emission rates, and primary organic concentrations is explored. Results show that temperature is likely to have a significant influence on SOA partitioning and resulting SOA concentrations. A 10 degrees C decrease in temperature is estimated to increase SOA yields by 20-150%, depending on the assumed vaporization enthalpy. In model simulations, high daytime temperatures tend to reduce SOA concentrations by 16-24%, while cooler nighttime temperatures lead to a 22-34% increase, compared to constant temperature conditions. Results suggest that currently available constant temperature partitioning coefficients do not adequately represent atmospheric SOA partitioning behavior. Air quality models neglecting the temperature dependence of partitioning are expected to underpredict peak SOA concentrations as well as mistime their occurrence. PMID- 11414010 TI - Indoor dose conversion coefficients for radon progeny for different ambient environments. AB - Inhaled progeny of 222Rn (radon progeny) are the most important source of irradiation of the human respiratory tract. Their attachment to atmospheric aerosols follows a well-established relationship between the activity size distribution (ASD) and the number size distribution. Recent studies have shown that indoor aerosols are derived primarily from outdoor sources, so it is pertinent to study the effects of different ambient environments on the indoor radon dose (in terms of the dose conversion coefficient or DCC, in units of mSv WLM-1). Commonly encountered ambient aerosols were studied here, which included the traffic-, urban-, and marine-influenced aerosols. The ASDs of attached radon progeny for all three studied ambient environments were well-represented by normal distributions. From these ASDs, the DCCs were calculated using the ICRP66 model and the scaled Yeh-Schum model. All other employed parameters were adopted from original references or authoritative reports. The DCCs for a nominal home calculated using the James model and the Yeh-Schum model were 12 and 8 mSv WLM-1, respectively. The DCCs were largest for urban-influenced ambient environments and smallest for marine-influenced ambient environments, and those for traffic influenced ambient environments were close to that for a nominal home. If we adopt the stochastic model, the probability of contracting radon-induced lung cancer by a person living with a marine-influenced ambient environment will be half that of a person living with an urban-influenced ambient environment. PMID- 11414011 TI - Behavior of sewage sludge-derived PAHs on pasture. AB - A field experiment was set up to investigate the retention of PAHs by pasture grass following sludge application. In addition to information on compound specific loss rates, the influence of meteorological variables and sludge-pasture contact time on the weathering process were investigated. The rates of loss were compound specific with half-lives ranging from < 1 to 10 d. The lighter PAHs were rapidly lost from the grass surface primarily by volatilization. For the intermediate and heavier PAHs, loss was slower with rain playing an important role in the weathering process. Growth dilution was also important in reducing the contaminant levels in the grass. The effect of sludge-pasture contact time prior to weathering by rain on the residual levels remains unclear. In terms of risk of exposure, there is the potential for persistent organic contaminants to be introduced into the grazing animal food chain if sewage sludge is applied to pasture land. PMID- 11414012 TI - Major ion and electrical potential distribution in soil under electrokinetic remediation. AB - To assess the effect of pore water chemistry on the contaminant removal rate, we monitored major ion concentrations in the pore water and the electrical potential distribution of a soil during electrokinetic remediation treatment. On application of the voltage, the soil near the anode became acidic due to the electrolytic generation of H+, and the acid region gradually spread toward the cathode. The major cation in the acid region was, however, not H+ but Al3+ that arose from the acid-induced dissolution of soil minerals, and it migrated very slowly toward the cathode. The measured pH and accompanying ion concentrations indicated that the anomalously slow migration of Al3+ was due to its precipitation-dissolution reaction at the acid front. The stagnancy of Al3+ increased the ionic concentration, flattened the electrical potential profile, and in turn, diminished electromigration in the acid region. This seems to be one of the causes of the relatively low removal rate of cationic and anionic contaminants in electrokinetic treatments. PMID- 11414013 TI - Seasonal and species differences in the air--pasture transfer of PAHs. AB - A field plot was established at a semirural site in the U.K. to investigate the atmospheric transfer of PAHs to different pasture species over the whole growing season. The PAHs displayed a range of partitioning behaviors in the atmosphere from exclusively gas phase to exclusively particle bound, resulting in different modes of deposition to the plant surface. The different pasture species had different plant and sward characteristics, e.g., leaf morphologies, yields, etc. For the majority of PAHs, the plant species displayed a seasonality in concentrations, with concentrations being higher in the winter than in the summer. For the lighter PAHs, this seasonality was absent with soil outgassing and/or summer sources of PAHs being implicated. Air-plant transfer factors (scavenging coefficients, with units m3/g dw) typically ranged between 4 and 52 during the summer, increasing to 8-88 during winter. Despite different plant and sward characteristics, the mixtures and concentrations of PAHs were similar for all the plant species. This indicates that there was little difference in the interception and retention behavior of the gas- and particle-phase PAHs. The implications of this for food chain transfer and air-vegetation modeling are discussed. PMID- 11414014 TI - Measurement of regulated and unregulated exhaust emissions from a lawn mower with and without an oxidizing catalyst: a comparison of two different fuels. AB - Relatively few emission characterization studies have been made on small engines used in garden equipment. The present investigation focuses on exhaust characterization from a lawn mower engine fueled with two different fuels in combination with and without an oxidizing catalyst. The compounds measured in the exhaust are carbon monoxide, hydrocarbons, nitrogen oxides, particulates, polycyclic aromatic hydrocarbons, methane, ethane, ethene, ethanol, and nitrous oxide. A significant reduction can be achieved by the use of a catalyst. By selection of the fuel, a significant reduction of certain carcinogenic compounds ("probably carcinogenic to humans" according to the IARC; benzo[a]pyrene and benzo[a]anthracene) may be achieved. The highest reduction improvement is achieved through the combination of an environmentally improved fuel, i.e., alkylate fuel, and a catalyst system. The data presented show that emissions from lawn mower engines are still relatively large although there is the potential for further improvements. PMID- 11414015 TI - The uptake of iron-55 by marine sediment, macroalgae, and biota following discharge from a nuclear power station. AB - Significant quantities of 55Fe, an activation product of stable iron, have been released into the environment following the atmospheric testing of nuclear weapons (mainly in the 1950s and 1960s) as well as through authorized discharges of radioactivity from nuclear power and reprocessing sites. Although some studies have been performed on the behavior of weapons' fallout-derived 55Fe in the environment and subsequent impact on humans, little has been published on the behavior of 55Fe released as a point source discharge from nuclear sites. This study presents data on the concentration and temporal variation of 55Fe in fucoid seaweeds, shellfish, crab, and lobster collected from Weymouth Bay and adjacent coastal areas, southern England. These areas have received authorized discharges of radionuclides originating from the operation of a now-decommissioned steam generating, heavy water-type reactor at AEE Winfrith. The highest activities of 55Fe are found associated with marine sediments collected near the discharge pipeline and a rapid decline occurs away from the pipeline. This is consistent with rapid sorption of 55Fe by the sediment, and the data show there is only limited reworking and remobilization. Activities of 55Fe in biota generally decreased over time, due to a reduction in the amount of 55Fe discharged. The variation of 55Fe activity, revealed from the monthly sampling of seaweed, does not reflect the short-term fluctuations seen in the patterns of discharged 55Fe activity. Although discharges of 55Fe from AEE Winfrith exceeded other radionuclides, the radiological impact on local seafood consumers is considerably less than for other key radionuclides such as 60Co and 65Zn but of comparable magnitude to the global average population dose arising from fallout-derived 55Fe. PMID- 11414016 TI - Fate of atmospherically deposited polycyclic aromatic hydrocarbons (PAHs) in Chesapeake Bay. AB - Factors controlling polycyclic aromatic hydrocarbon (PAH) distributions in southern Chesapeake Bay surface sediments were investigated with samples collected seasonally from five sites. Principal components analysis (PCA) suggests unique sources (combustion vs petroleum) and partitioning differences (volatile vs particle reactive) among PAHs, but a common mode of entry for these contaminants to Chesapeake Bay sediments. The fractional concentrations of all PAHs in Bay sediments, with the exception of perylene, were positively correlated with their atmospheric dry deposition fluxes (Fdry) to the Bay indicating that atmospheric deposition of aerosol-bound PAHs to the watershed controls their influx to Bay sediments. Overall, PAH concentrations in Chesapeake Bay sediments were well explained by a multiple-regression model (r2 = 0.88, p = 0.0001) with Fdry and sediment total organic carbon (TOC) content explaining most of the variance (57% and 43%, respectively). However, for many PAHs gas exchange across the air-water interface is of similar or greater magnitude even when Fdry is scaled to the watershed surface area. The fate of PAHs input to the Bay from gas deposition was determined to be uptake and metabolism within the aquatic food web rather than deposition to sediments. PMID- 11414017 TI - Detection of alkaline ultrafine atmospheric particles at Bakersfield, California. AB - Two collected micro-orifice uniform deposit impactors (MOUDIs) and a filter-based sampler were used to measure the size distribution and chemical composition of atmospheric particulate matter at Bakersfield, CA, between January 14 and 23, 1999. The peak number concentration of airborne ultrafine particles measured was 1.45 x 10(11) m-3, which is a factor of approximately 3 higher than the peak airborne ultrafine-particle number concentration measured previously in Pasadena, CA. Chemical analysis revealed that airborne ultrafine particles (Dp < 0.1 micron) at Bakersfield contained significant amounts of water-soluble species, including calcium, sodium, ammonium ion, nitrate, and sulfate. Other chemical species detected in the ultrafine size range included potassium, iron, copper, zinc, and strontium. A balance of aqueous ions showed that ultrafine particles were alkaline in nature with calcium acting as the dominant cation. Bulk samples of airborne particles with diameter less than 2.0 microns (PM2.0) were essentially neutral, but particle acidity was found to be a strong function of particle size. The results of this experiment suggest that areas deep in the human lung that preferentially collect particles in the ultrafine size range could be exposed to locally acidic or alkaline conditions even if the integrated airborne particle complex is essentially neutral. PMID- 11414018 TI - Heterogeneous reaction of NO2 on diesel soot particles. AB - Soot particles were collected from a diesel engine using a procedure that realistically mimics exhaust gas conditions in tailpipes and during dilution at room temperature. After being sampled, the particles were exposed to NO2 concentrations and relative humidity in ranges relevant for the troposphere using 13N as tracer. Gas-phase nitrous acid(HONO) and irreversibly bound (i.e., chemisorbed) species were the main reaction products with initial yields of 80 90% and about 10%, respectively. Neither NO nor HNO3 were detectable. The HONO formation increased with increasing engine load (i.e., with a decreasing air to fuel ratio, lambda). The reaction rates of HONO and chemisorbed NO2 increased with increasing NO2 concentration and did not depend on relative humidity. At the beginning of reaction, the uptake coefficient averaged over 3 min ranged from 5 x 10(-6) to 10(-5) for NO2 concentrations between 2 and 40 ppb. The HONO formation rates decreased with time, indicating consumption of reactive surface species, while the chemisorption rates remained almost constant. The total HONO formation potential of the particles was estimated to about 1.3 x 10(17) molecules/mg of diesel soot or to about 4.7 mg/kg of diesel fuel, indicating that the reaction between NO2 and diesel soot particles does not provide a significant secondary HONO source in the atmosphere. A Langmuir-Hinshelwood type reaction mechanism was proposed that adequately describes the observed results and also allows discussing important general features of reactions on soot. PMID- 11414019 TI - Effects of surface type and relative humidity on the production and concentration of nitrous acid in a model indoor environment. AB - A nested chamber design was constructed for the purpose of studying parameters that affect indoor air chemistry. Experiments were conducted in this system to investigate the effects of three surface types (Teflon, wallpaper, and carpet) and two levels of relative humidity (50% and 70% RH) on the formation of gas phase nitrous acid (HONO) through the heterogeneous reaction of nitrogen dioxide (NO2) with sorbed water vapor. The results of this investigation show that, compared with Teflon surfaces, carpet made of synthetic fibers increased the NO2 surface removal rate by nearly an order of magnitude and resulted in higher peak HONO concentrations. The results also suggest that the capacity of a surface to sorb water will determine if HONO is released from that surface after the NO2 source has been turned off and the heterogeneous reaction between NO2 and sorbed water is no longer significant. Vinyl-coated wallpaper was found to release HONO for prolonged periods of time after the NO2 source was turned off at both 50% and 70% RH whereas Teflon was found to do so only at 70% RH. The results of this investigation also demonstrate the utility of the nested chamber design in investigating indoor air chemistry. PMID- 11414020 TI - Predicting the wettability of quartz surfaces exposed to dense nonaqueous phase liquids. AB - A model incorporating electrostatic and van der Waals forces was used to predict the wetting behavior of several systems containing a quartz surface, water, and a dense nonaqueous phase liquid (DNAPL). The disjoining pressure between the quartz/water and a water/DNAPL interface as a function of their separation was calculated from a full solution of the nonlinear Poisson-Boltzmann equation allowing for charge regulation. For the pH regime associated with a positive pressure barrier, the quartz surface was predicted to be water wetting. The model takes into account the charging behavior of an isolated DNAPL/water interface as inferred from electrophoretic mobility data. It provides excellent predictions for four coal tar and creosote samples tested, where the quartz was oil-wetting below pH 5 and water-wetting at higher pH values. In a fifth sample, the transition occurred above pH 7, which indicates that mechanisms other than electrostatic interaction control the wetting behavior in this sample. PMID- 11414021 TI - Thiosulfate enhances silver uptake by a green alga: role of anion transporters in metal uptake. AB - Short-term (< 1 h) silver uptake by the green alga Chlamydomonas reinhardtii was measured in the laboratory in defined inorganic media in the presence or absence of ligands (chloride and thiosulfate). In contradiction to the free-ion model of metal uptake, silver accumulation by the alga proved to be sensitive to the choice of ligand used to buffer the free silver concentration. For a low fixed free Ag+ concentration of 10 nM, silver uptake in the presence of thiosulfate (0.11 microM) was 2x greater than in the presence of chloride (4 mM). When sulfate was removed from the exposure medium (i.e., 81 microM-->0 microM), silver uptake in the presence of thiosulfate was even more markedly enhanced (more than 4x greater than in the presence of chloride). Varying the sulfate concentration in the exposure medium only affected silver uptake if thiosulfate was present. We conclude that silver-thiosulfate complexes are transported across the plasma membrane via sulfate/thiosulfate transport systems and that sulfate acts as a competitive inhibitor of this uptake mechanism. PMID- 11414022 TI - Chromium(VI) reduction by hydrogen sulfide in aqueous media: stoichiometry and kinetics. AB - The objective of this work was to investigate the reaction stoichiometry, kinetics, and mechanism for Cr(VI) reduction by hydrogen sulfide in the aqueous phase. Batch experiments with excess [Cr(VI)] over [H2S]T indicated that the molar amount of sulfide required for the reduction of 1 M Cr(VI) was 1.5, suggesting the following stoichiometry: 2CrO4(2-) + 3H2S + 4H+-->2Cr(OH)3(s) + 3S(s) + 2H2O. Further study with transmission electron microscopy (TEM) and energy-dispersive X-ray spectroscopy (EDS) confirmed that chromium hydroxide and elemental sulfur were the stable products. The kinetics of Cr(VI) reduction by hydrogen sulfide was measured under various initial concentrations of Cr(VI) and sulfide as well as pH values controlled by HEPES, phosphate, and borate buffers. Results showed that the overall reaction was second-order, i.e., first-order with respect to Cr(VI) and first-order to sulfide. The reaction rate increased as pH was decreased, and the pH dependence correlated well with the fraction of fully protonated sulfide (H2S) in the pH range of 6.5-10. The nature of buffers did not influence the reaction rate significantly in the homogeneous system. The reaction kinetics could be interpreted by a three-step mechanism: formation of an inner sphere chromate-sulfide intermediate complex ((H2O4CrVIS)2-), intramolecular electron transfer to form Cr(IV) species, and subsequent fast reactions leading to Cr(III). PMID- 11414023 TI - Influence of clay minerals on the hydrolysis of carbamate pesticides. AB - Using batch experiments, we investigated the influence of clay minerals (montmorillonite, beidellite, illite, and vermiculite) on the hydrolysis of five carbamate pesticides: carbosulfan, carbofuran, aldicarb, pirimicarb, and chlorpropham. Compared to the other minerals, montmorillonite had the strongest influence on the hydrolysis of these carbamates. Montmorillonite enhanced the hydrolysis of carbosulfan and aldicarb. In contrast, the hydrolysis of chlorpropham was inhibited by montmorillonite, probably because of its strong adsorption on montmorillonite. The hydrolysis of pirimicarb was not affected by montmorillonite. The presence of organic substances, phosphate, and fluoride in suspensions decreased the catalytic activity of montmorillonite. Surface acidity of montmorillonite and/or formation of surface chelates are probably the key factors of surface catalysis in the case of the hydrolysis of carbosulfan. PMID- 11414024 TI - Chemical analysis of diesel engine nanoparticles using a nano-DMA/thermal desorption particle beam mass spectrometer. AB - Diesel engines are known to emit high number concentrations of nanoparticles (diameter < 50 nm), but the physical and chemical mechanisms by which they form are not understood. Information on chemical composition is lacking because the small size, low mass concentration, and potential for contamination of samples obtained by standard techniques make nanoparticles difficult to analyze. A nano differential mobility analyzer was used to size-select nanoparticles (mass median diameter approximately 25-60 nm) from diesel engine exhaust for subsequent chemical analysis by thermal desorption particle beam mass spectrometry. Mass spectra were used to identify and quantify nanoparticle components, and compound molecular weights and vapor pressures were estimated from calibrated desorption temperatures. Branched alkanes and alkyl-substituted cycloalkanes from unburned fuel and/or lubricating oil appear to contribute most of the diesel nanoparticle mass. The volatility of the organic fraction of the aerosol increases as the engine load decreases and as particle size increases. Sulfuric acid was also detected at estimated concentrations of a few percent of the total nanoparticle mass. The results are consistent with a mechanism of nanoparticle formation involving nucleation of sulfuric acid and water, followed by particle growth by condensation of organic species. PMID- 11414025 TI - Abiotic transformation of perchloroethylene in homogeneous dithionite solution and in suspensions of dithionite-treated clay minerals. AB - The reductive dechlorination of perchloroethylene (PCE) in homogeneous solutions of dithionite and at the surfaces of dithionite-citrate-bicarbonate (DCB) treated ferruginous smectite and Na-montmorillonite was studied. Transformation products of PCE identified in dosed dithionite-treated samples included TCE, DCE, 1,1,2 trichloroethane (TCA), 1,1-dichloroethane (DCA), chloroacetylene, acetylene, ethene, and ethane. The decomposition of dithionite to sulfate yielded both protons and electrons necessary for hydrodechlorination (hydrogenolysis) of PCE. Dithionite treatment of the Fe-poor Na-montmorillonite enhanced reductive dechlorination of PCE relative to dithionite-treated Fe-rich ferruginous smectite, within the range of 11.5-137.8 mM dithionite. For the same dithionite concentration, the kinetics of the heterogeneous reactions of PCE was generally faster than that of the homogeneous reaction, and higher concentrations of TCE were measured in the heterogeneous reactions. Interestingly, increases in the mass of the clay minerals used, the Fe2+ content in the clay mineral structure, or the dithionite concentration used did not necessarily enhance the abiotic transformation of PCE, as would otherwise be predicted. The most efficient reductive dechlorination of PCE was observed with 0.5% clay (m/v) treated with 34.5 mM dithionite buffered at pH 8.5. The solid-state transfer of electrons to surfaces and edges, rather than the redox capacity, limited the dechlorination of PCE by reduced ferruginous smectite and/or suspensions containing a higher clay mass. The greater reactivity of dithionite-reduced montmorillonite than similarly treated ferruginous smectite is attributed to (i) the well-documented layer collapse and aggregation of chemically reduced clays that increases with the clay's iron content, (ii) the location of solid-phase Fe2+ in the reduced clay mineral and whether it is accessible or inaccessible for reaction with PCE at the mineral edges and surfaces where the reactions are thought to occur, and (iii) the greater swellability of montmorillonite versus ferruginous smectite. The faster dechlorination rate of PCE observed with dithionite-reduced Fe-poor montmorillonite than similarly reduced iron-rich ferruginous smectite suggests that the use of dithionite barriers for in-situ treatment of chlorinated solvent plumes should not be limited to aquifers with Fe-rich sediments. PMID- 11414026 TI - Aqueous geochemical and surface science investigation of the effect of phosphate on pyrite oxidation. AB - Aqueous geochemical and surface science techniques were used to investigate the effect of phosphate on the oxidation of crushed and (100) pyrite. Studies showed that the presence of phosphate in solution significantly impeded the oxidation rate of crushed pyrite at pH values equal to or greater than 4. At pH 3, the presence of phosphate had almost no experimentally discernible effect on pyrite oxidation. X-ray photoelectron spectroscopy (XPS) studies showed that phosphate, at pH values greater than 4, became irreversibly bound to an Fe(3+)-bearing product on the pyrite surface during the oxidation process. Once bound to this region on the pyrite surface, the adsorbed phosphate inhibited further oxidation of the pyrite (based on XPS determinations of sulfur and iron oxide product concentrations) under our experimental conditions. These results suggested that the rate of pyrite oxidation in the absence of phosphate was facilitated at or near Fe(3+)-bearing oxidation phases on the surface. Phosphate bound on iron(III) oxide product either prevents O2 adsorption on this phase or electronically modifies these surface regions, but in either case it inhibits electron transfer from pyrite-Fe2+ sites to molecular O2. PMID- 11414027 TI - Reduction of haloacetic acids by Fe0: implications for treatment and fate. AB - To predict the fate of haloacetic acids (HAAs) in natural or engineered systems, information is needed concerning the types of reactions that these compounds undergo, the rates of those reactions, and the products that are formed. Given that many drinking water distribution systems consist of unlined cast iron pipe, reactions of HAAs with elemental iron (Fe0) may play a role in determining the fate of HAAs in these systems. In addition, zerovalent iron may prove to be an effective treatment technology for the removal of HAAs from chlorinated drinking water and wastewater. Thus, batch experiments were used to investigate reactions of four trihaloacetic acids, trichloroacetic acid (TCAA), tribromoacetic acid (TBAA), chlorodibromoacetic acid (CDBAA), and bromodichloroacetic acid (BDCAA), with Fe0. All compounds readily reacted with Fe0, and investigation of product formation and subsequent disappearance revealed that the reactions proceeded via sequential hydrogenolysis. Bromine was preferentially removed over chlorine, and TBAA was the only compound completely dehalogenated to acetic acid. In compounds containing chlorine, the final product of reactions with Fe0 was monochloroacetic acid. Halogen mass balances were 95-112%, and carbon mass balances were 62.6 112%. The pseudo-first-order rate constants for trihaloacetic acid degradation were as follows: BDCAA (10.6 +/- 3.1 h-1) > CDBAA (1.43 +/- 0.32 h-1) approximately TBAA (1.41 +/- 0.28 h-1) >> TCAA (0.08 +/- 0.02 h-1). PMID- 11414028 TI - What is environmental research? PMID- 11414029 TI - Temporal trends, temperature dependence, and relative reactivity of atmospheric polycyclic aromatic hydrocarbons. AB - Atmospheric polycyclic aromatic hydrocarbon (PAH) concentrations have declined appreciably between 1992 and 1997 at an urban background site in Birmingham, U.K. In contrast, no decline is evident in the city center between 1994 and 1997. Although most PAHs display statistically significant negative correlation with temperature, so does NOx, for which traffic (a nonseasonal activity) is the major source, and for which the negative correlation with temperature reflects seasonal boundary layer depth variations. When concentrations of PAHs divided by NOx were plotted against temperature, no significant relationship was detected for any PAH, except fluorene, phenanthrene, and fluoranthene. For these PAHs, the relationship was positive, suggesting volatilization from surfaces may be appreciable. For samples collected simultaneously at the city center and urban background sites, greater negative temperature-dependence was observed at the latter location. Although this may be partly due to the fact that the enhanced reactivity of PAHs at higher temperatures exerts a greater influence at sites more distant from emissions; the dichotomy in temperature-dependent behavior and temporal trends may also be due to city center concentrations being "buffered" by volatilization from surfaces to a greater extent than those at the urban background site. PMID- 11414030 TI - Alkyl bromides as mechanistic probes of reductive dehalogenation: reactions of vicinal dibromide stereoisomers with zerovalent metals. AB - Whether reductive dehalogenation proceeds via a one- or a two-electron mechanism has been suggested to affect product distributions, hence potentially influencing the success of engineered treatment systems. In this work, we explore vicinal dibromide stereoisomers as "probes" of the concertedness of electron transfer in reduction by aqueous suspensions of iron and zinc metal. Dibromides consisted of 2,3-dibromopentane (diBP) stereoisomers and (+/-)-1,2-dibromo-1,2-diphenylethane. All dibromides reacted with metals to give the same E:Z ratio of olefins observed during dehalogenation by iodide (a two-electron reductant). Reduction by Cr(II) (a one-electron reductant) yielded distinctly different proportions of E and Z olefins. Although this might be construed as evidence that metals function as two electron reductants, high stereo-specificity was also obtained for reduction of diBPs by Fe(II) adsorbed to goethite, a presumed one-electron reductant; this can be explained by two single-electron transfers in rapid succession, facilitated by the locally elevated concentration of reducing equivalents at the oxide-water interface. The results suggest that reduction of alkyl halides by metals is not likely to produce free radicals that persist long enough to undergo radical radical coupling or hydrogen-atom abstraction from minor dissolved constituents. Apparent free-radical coupling products are more likely to result from (possibly surface-bound) organometallic intermediates. PMID- 11414031 TI - Bush's federal budget goes light on R&D. PMID- 11414032 TI - Incipient erosion of biostabilized sediments examined using particle-field optical holography. AB - Laser holography allows images of three-dimensional space at ultra-high resolution to be recorded onto photographic plates. Recorded scenes can be "replayed" with a second laser beam into free space and optically "interrogated" using either a microscope or a camera by sequentially focusing on increasing distances from the hologram in the field of view (optical sectioning). From these sections, information on the relative locations and orientation in space of suspended particles as well as the morphology of particles can be obtained. This paper examines the utility of "in-line" laser holography to discriminate the size and the morphology of sand particles eroded under turbulent shear flow during benthic sediment transport. The influence of a commercially available adhesive polymer (xanthan gum, derived from the bacterium Xanthomonas campestris) on sediment stability and resuspended particle morphology is described. The major implications for carbon and sediment cycling within estuaries are highlighted. PMID- 11414033 TI - A molecular topology approach to predicting pesticide pollution of groundwater. AB - Various models have proposed methods for the discrimination of polluting and nonpolluting compounds on the basis of simple parameters, typically adsorption and degradation constants. However, such attempts are prone to site variability and measurement error to the extent that compounds cannot be reliably classified nor the chemistry of pollution extrapolated from them. Using observations of pesticide occurrence in U.S. groundwater it is possible to show that polluting from nonpolluting compounds can be distinguished purely on the basis of molecular topology. Topological parameters can be derived without measurement error or site specific variability. A logistic regression model has been developed which explains 97% of the variation in the data, with 86% of the variation being explained by the rule that a compound will be found in groundwater if 6 chi Pv < 0.55. Where 6 chi p is the sixth-order molecular path connectivity. One group of compounds cannot be classified by this rule and prediction requires reference to higher order connectivity parameters. The use of molecular approaches for understanding pollution at the molecular level and their application to agrochemical development and risk assessment is discussed. PMID- 11414034 TI - Source apportionment of urban particulate aliphatic and polynuclear aromatic hydrocarbons (PAHs) using multivariate methods. AB - Samples of organic aerosol were collected in Santiago de Chile. An activated charcoal diffusion denuder was used to strip out organic vapors prior to particle collection. Both polynuclear aromatic hydrocarbons (PAHs) and aliphatic hydrocarbons were determined using gas chromatography/mass spectrometry (GC/MS). Organic particle sources were resolved using both concentration diagnostic ratios and multivariate methods such as hierarchical cluster analysis (HCA) and factor analysis (FA). Four factors were identified based on the loadings of PAHs and n alkanes and were attributed to the following sources: (1) high-temperature combustion of fuels; (2) fugitive emissions from oil residues; (3) biogenic sources; and (4) unburned fuels. Multilinear regression (MLR) analysis was used to determine emission profiles and contributions of the sources. The reconstructed concentrations of particle phase aliphatic and polynuclear aromatic hydrocarbons were in good agreement (R2 > 0.70) with those measured in Santiago de Chile. PMID- 11414035 TI - Plutonium partitioning to colloidal and particulate matter in an acidic, sandy sediment: implications for remediation alternatives and plutonium migration. AB - Plutonium partitioning within a bulk, freshwater sediment and to specific size fractions of the sediment was determined. Fission track analysis was used to observe the spatial heterogeneity of fissile isotopes of Pu and U in the sediment. For the bulk sediment, a six-step sequential extraction scheme was used to elucidate Pu partitioning. Although no direct Pu speciation is obtained from our sequential extraction scheme, this study demonstrates that the partitioning information obtained is more useful for evaluating ex-situ remediation treatments than information obtained from complete digestion and analysis of the bulk sediment. The majority of 238Pu and 239 + 240Pu appear to be partitioned in the oxidizable fraction, suggesting that Pu is primarily associated with organic matter in the bulk sediment or may exist as a discrete, oxidizable phase. By varying filter pore size used to separate the sequential extraction leachate solution from the remaining solid phase, a fraction of 238Pu associated with colloidal material was observed, and chemical evidence suggests that this colloidal material is relatively refractory. Pu partitioning to various size fractions of the bulk sediment was also compared to the percent organic carbon present in those size fractions. Interestingly, little correlation was observed between the percentage of organic carbon and concentrations of Pu isotopes in the various size fractions, although differences were observed in the distributions of 238Pu versus 239 + 240Pu in the size fractions. These results suggest that other sediment phases may also be important for Pu partitioning. Our observations are described in the context of feasibility of various remediation options. PMID- 11414036 TI - Evaluation of media and derivatization chemistry for six aldehydes in a passive sampler. AB - We evaluated the GMD passive sampler for its suitability to measure six aldehydes over a 7-d period in population exposure studies. The six target aldehydes were formaldehyde, acetaldehyde, acrolein, crotonaldehyde, glyoxal, and methylglyoxal. The GMD sampler contains a silica gel-impregnated cellulose pad coated with 2,4 dinitrophenylhydrazine (DNPH) hydrochloride. This agent reacts with formaldehyde to form a hydrazone that is quantified with a high-performance liquid chromatograph. The GMD sampler was tested for background contamination and aldehyde recoveries after 0, 1, and 7 d of storage. Results indicated that the GMD monitor, as currently manufactured, is suitable for shorter-term sampling (up to 24 h) of formaldehyde and acetaldehyde. It is however not acceptable for sampling of acetaldehyde, acrolein, crotonaldehyde, glyoxal, and methylglyoxal over a 7-d exposure period due to the chemical reactions on the silica gel impregnated cellulose pad. Glyoxal- and methylglyoxal-DNPH derivatives formed on the cellulose and Teflon-coated glass fiber pads that had been prepared with glycerol under acidic and oxidative conditions. Acrolein- and crotonaldehyde-DNPH derivatives diminish through the reverse reaction of the DNPH derivatives to form free aldehydes under acidic conditions. We showed that the unknown reaction products of acrolein and crotonaldehyde derivatives were not pyrazolines but probably resulted from E/Zisomerization. These conversion reactions are favored in acidic conditions present in either the derivatization solution or the collection medium. The most consistent recovery was obtained on glass fiber pads. In particular, recoveries of crotonaldehyde- and acrolein-DNPH derivatives were increased through the use of a pH 4 buffered derivatization solution. These chemical instability problems were overcome by using a pH 4 buffer (citric acid/sodium citrate) and an alternative hygroscopic agent (1,3-butanediol) in the DNPH derivatization solution. Results with DNPH derivatives from these spiking experiments were further confirmed with gas-phase spiking experiments. We determined the optimal acidity, buffer solution, and concentrations of the buffer solution and 1,3-butanediol for the DNPH derivatization solution. This new formulation of the DNPH derivatization solution can be used for collection of the six target aldehydes over a 7-d sampling period. PMID- 11414037 TI - Field evaluation of a method for estimating gaseous fluxes from area sources using open-path Fourier transform infrared. AB - This paper describes results from the first field experiment designed to evaluate a new approach for quantifying gaseous fugitive emissions of area air pollution sources. The approach combines path-integrated concentration data acquired with any path-integrated optical remote sensing (PI-ORS) technique and computed tomography (CT) technique. In this study, an open-path Fourier transform infrared (OP-FTIR) instrument sampled path-integrated concentrations along five radial beam paths in a vertical plane downwind from the source. A meteorological station collected measurements of wind direction and wind speed. Nitrous oxide (N2O) was released from a controlled area source simulator. The innovative CT technique, which applies the smooth basis function minimization method to the beam data in conjunction with measured wind data, was used to estimate the total flux from the simulated area source. The new approach estimates consistently underestimated the true emission rates in unstable atmospheric conditions and agreed with the true emission rate in neutral atmospheric conditions. This approach is applicable to many types of industrial areas or volume sources, given the use of an adequate PI ORS system. PMID- 11414038 TI - Resolution of GC-MS data of complex PAC mixtures and regression modeling of mutagenicity by PLS. AB - The present work describes a strategy to predict the mutagenicity of very complex mixtures of polycyclic aromatic compounds (PAC) from gas chromatography-mass spectrometry (GC-MS) patterns of the mixtures, each containing 260 compounds on average. The mixtures, 13 organic extracts of exhaust particles, were characterized by full scan GC-MS. The data were resolved into peaks and spectra for individual compounds by an automated curve resolution procedure. Similarity between spectra was evaluated for peaks that appeared within a time interval of 4 min, using a similarity index of 0.8 to ascertain that the same compound was represented by the same variable name (retention time) in all samples. The resolved chromatograms were integrated, resulting in a predictor matrix of size 13 x 721, which was used as input to a multivariate regression model. Partial least-squares projections to latent structures (PLS) were used to correlate the GC-MS chromatograms to mutagenicity as measured in the Ames Salmonella assay. The best model (high r2 and Q2) was obtained with 52 variables. These variables covary with the observed mutagenicity, and may subsequently be identified chemically. Furthermore, the regression model can be used to predict mutagenicity from GC-MS chromatograms of other organic extracts. PMID- 11414039 TI - Predicting soil-water partitioning of polycyclic aromatic hydrocarbons and polychlorinated biphenyls by desorption with methanol-water mixtures at different temperatures. AB - We evaluated a method to determine organic carbon-normalized soil-water partition coefficients (Koc) of 20 PAHs and 12 PCBs by desorption in the presence of a cosolvent (methanol fractions of 0.1-0.9) and at different temperatures (20-80 degrees C). The Koc values, the deviation factor from ideal sorption alpha, and the desorption enthalpies delta Hdes were estimated by nonlinear regression of log Koc on the methanol fractions and on T. The Koc values of individual compounds varied up to a factor of 100 among the studied 11 urban soils. The calculated alpha and delta Hdes of individual compounds varied considerably among the soils (coefficients of variation 5-20% and 20-30%, respectively), alpha increased with increasing hydrophobicity of the compounds. A sequential extraction with four temperature/methanol fraction combinations followed by a nonlinear regression allowed for the direct determination of the Koc, alpha, and delta Hdes. The use of less temperature/methanol fraction combinations requires a suitable estimation of alpha and delta Hdes, as their choice may change the obtained Koc values by up to a factor of 10. The proposed method is suitable for a routine determination of Koc values of PAHs and PCBs for small soil samples (2 6 g) and low concentrations (down to 0.3 mg kg-1 of sigma 20 PAHs and 1.2 micrograms kg-1 of sigma 12 PCBs). PMID- 11414040 TI - Analysis of semivolatile organic compounds in atmospheric aerosols by direct sample introduction thermal desorption GC/MS. AB - A technique for identifying trace amounts of semivolatile organic compounds in atmospheric aerosols and in the NIST Urban Dust standard (SRM1649a) is presented. The technique is based on direct sample introduction (DSI) of small samples followed by thermal desorption in a conventional GC injector. The method enables injection of both solid and liquid samples. Validation of the method, including quantitative determination of EPA-targeted polycyclic aromatic hydrocarbons (PAHs), as well as the reproducibility and recovery efficiency tests are presented. The advantages of using aluminum foil as sampling filter are also discussed. Determination of different classes of compounds such as quinolines, methylquinoline isomers, PAHs, and n-monocarboxylic acids in the ambient size segregated aerosol sample is also performed. The method was directly applied to the determination of C6-C16 n-monocarboxylic acids, eliminating the need for a complex sample preparation procedure. The small quantities needed for the analysis as well as the lack of complicated sample preparation steps enable fast characterization of semivolatile organic species present in time-resolved or size segregated aerosol samples. Thus, this method can potentially be employed for air quality monitoring and field measurements as well as for fast screening of the organic content of ambient particles. PMID- 11414041 TI - Capping efficiency for metal-contaminated marine sediment under conditions of submarine groundwater discharge. AB - Theoretical estimations and laboratory studies suggest that capping can effectively retard contaminant transport from sediments under undisturbed conditions. However, contaminated near-shore areas, commonly selected as capping sites, are frequently subjected to submarine groundwater discharge (SGD). Column experiments were set up in the laboratory to simulate metal transport through sediment and capping material in the presence and absence of SGD. In the absence of SGD, capping enhanced Mo flux and initial Mn flux while having no effect in retarding Fe flux, presumably due to altered redox conditions. This effect was more pronounced in the presence of SGD (4.7 x 10(-4) m/h specific discharge). Capping enhanced Cd flux and initial fluxes of Ni, Cu, and Zn under conditions of simulated SGD, which may be caused by co-transport with Mn and Fe and oxidation of sulfide. Capping retarded Cr and Pb fluxes and steady-state Ni, Cu, Zn, and Fe fluxes in the presence of simulated SGD. However, capping efficiency decreased relative to that with no SGD. Elevated Mn concentration was detected at the capping surface with simulated SGD. Results indicate that advective flow may lead to significantly higher metal fluxes than those under undisturbed conditions. PMID- 11414042 TI - Removal of phenols from aqueous streams by the cloud point extraction technique with oxyethylated methyl dodecanoates as surfactants. AB - New oxyethylated methyl dodecanoates with average degrees of oxyethylation (n) ranging between 5 and 14 (denoted OMD-n) were considered as nonionic surfactants for the removal of phenol, 4-methylphenol, and 4-nitrophenol from aqueous streams by the cloud point extraction technique. The cloud point (denoted CP) strongly increases with the average degree of oxyethylation n but is independent of OMD-n concentration, except at low concentration (i.e., < 10 g.L-1). CP can be related to the surfactant hydrophilicity by the following equation: CP(degree C) = 20.27 x HLBG - 245.8 (above 10 g.L-1 and in the absence of electrolyte), where HLBG refers to the OMD-n hydrophile-lipophile balance in Griffin's scale. The presence of electrolyte such as NaCl strongly decreases the cloud point [delta CP/delta [NaCl] approximately -(9 +/- 1) degrees C-M-1]. Many parameters influence the extraction of phenols in OMD-n-rich phases, including the characteristics of phenols (hydrophobicity, hydrogen-bond acidity, hydrogen-bond basicity, etc.), the average degree of oxyethylation of OMD-n (i.e., n), the concentration of electrolyte, the settling temperature (Tset), and overheating (delta T = Tset - CP). The distribution coefficients typically range between 20 and 100. Finally, the kinetics of the separation process is limited by the slow coalescence of the fine droplets of surfactant-rich phase formed above CP. PMID- 11414043 TI - Nitrous oxide emissions from aerated composting of organic waste. AB - The composting of high organic content wastes has been shown to produce nitrous oxide (N2O). This study was initiated to investigate the mechanisms of N2O emissions from aerated composting and to determine the optimal operational conditions that minimize N2O emissions. The results of our experiment in laboratory-scale composters showed that more than 95% of N2O was produced during the later period of composting when readily available carbon sources had been depleted. Significant increases in N2O emission after nitrite (NO2-) addition, and good NO(2-)-N2O correlation, indicates that N2O was transformed from NO2-. Extremely high N2O generation was observed after NO2- addition in the presence and absence of composted cattle manure. This suggests an identical mechanism for N2O production in both treatments. However, the addition of composted cattle manure resulted in an earlier initiation of the main N2O generation period. Intermittent feeding of fresh food waste postponed the main N2O generation period, and reduced the mass-based N2O emissions by 20%. PMID- 11414044 TI - Kinetics of removal of carbonyl sulfide by aqueous monoethanolamine. AB - The kinetics of the reaction between carbonyl sulfide and aqueous monoethanolamine (MEA) were studied over a range in temperature (298-348 K) and amine concentrations (5-20 wt%) using a wetted-sphere absorber. The experimental data were interpreted using a zwitterion mechanism. The key physicochemical properties needed to interpret the data are the solubility and diffusivity of COS in the aqueous amine solution. These properties were estimated using the N2O analogy method. Experimental values of N2O solubility were correlated using an extended scaled-particle model, and the measured N2O diffusion coefficients were correlated using a modified Stokes-Einstein equation. Solution densities and viscosities were also measured and correlated in this work. On the basis of the zwitterion mechanism whose rate-limiting step was the deprotonation of a zwitterion, the Arrhenius relationship between the third-order rate constant and the temperature was well correlated with an absolute mean deviation of 0.3%. It could be thus concluded that the overall reaction rate was first-order in the COS concentration and second-order in the MEA concentration. PMID- 11414045 TI - Sonolytic, photolytic, and photocatalytic decomposition of atrazine in the presence of polyoxometalates. AB - Aqueous solutions of atrazine [2-chloro-4-(isopropylamino)-6-(ethylamino)-s triazine] (CIET) decompose upon illumination with a low-pressure Hg-arc lamp (254 nm). However, no decomposition takes place with lambda > 300 nm. On the other hand, addition of polyoxometalates (POM), PW12O40(3-) or SiW12O40(4-), into a solution of atrazine photodecomposes the substrate within a few minutes (cutoff fiter 320 nm). Ultrasound (US) treatment also decomposes aqueous solutions of atrazine within a few minutes. Both methods, sonolysis and photolysis with POM, give common intermediates, namely, 2-hydroxy-4-(isopropylamino)-6-amino-s triazine (OIET), 2-chloro-4-(isopropylamino)-6-amino-s-triazine (CIAT), 2-chloro 4-amino-6-(ethylamino)-s-triazine (CAET), 2-hydroxy-4,6-diamino-s-triazine (OAAT), and 2-hydroxy-4-hydroxy-6-amino-s-triazine (OOAT) among others. The final products for both methods, US and photolysis with POM, were cyanuric acid (OOOT), NO3-, Cl-, CO2, and H2O. OOOT showed no signs of decomposition by sonication and/or photolysis with POM. It also resisted degradation upon photolysis with plain UV light (254 nm). However, it has been reported to decompose upon photolysis with lambda > 200 nm. Combination of US and photolysis with POM produces only a cumulative effect. PMID- 11414046 TI - Degradation of bisphenol A in water by TiO2 photocatalyst. AB - The photocatalytic degradation of bisphenol A (BPA), a representative endocrine disruptor, was carried out in TiO2 aqueous suspension. The main purposes were to confirm the total mineralization of BPA and to evaluate the estrogenic activity in the treated water during the photocatalytic reaction. An initial BPA concentration of 175 microM in water was totally degraded to carbon dioxide by TiO2-photocatalyzed reactions under UV irradiation of 10 mW cm-2 for 20 h. Four HPLC peaks indicating intermediate products appeared in chromatograms monitored at 275 nm, but the heights relative to that of the initial BPA were very low, at most 0.04 in the time period 5-10 h after the start of UV irradiation. All of the peaks finally disappeared after 20 h. For the treated water, the transcriptional estrogenic activity in response to human estrogen receptor in a yeast hybrid assay decreased drastically to less than 1% of the initial BPA's activity within 4 h. On the basis of these results, we conclude that TiO2 photocatalysis could be a useful technology for the purification of water containing BPA without generating any serious secondary pollution. PMID- 11414047 TI - Catalytic reduction of NO with NH3 over carbons impregnated with Cu and Fe. AB - Selective catalytic reduction (SCR) with NH3 remains as one of the most effective technologies controlling NOx emission from stationary sources. At low temperatures carbon exhibits higher catalytic activity in NOx reduction than conventional catalysts made of metal oxides or zeolites. In the present work, the rate of catalytic NO reduction conducted at 110-200 degrees C over mineral matter free carbon particles was found to be low, contrary to the findings reported in the literature. The activity of the carbon was significantly enhanced from impregnating it with nitrates of Cu and Fe. The carbon impregnated with Cu shows the highest activity. The affinity of these carbon catalysts toward NH3 and NO, as well as the tendency of the impregnated metal oxides toward reduction by carbon and H2, has an order identical to that of their activity in NO reduction. The high activity of the metal-loaded carbons was related to the combined catalytic effects of the carbon and the metal. It has been suggested on the basis of this study that the metal-carbon complexes may serve as the catalytic centers for oxygen transference from NO to NH3, which is essentially slow in the absence of the metal. PMID- 11414048 TI - Chlorothalonil and its 4-hydroxy derivative in simple quartz sand soils: a comparison of sorption processes. AB - Quartz sandy soils from Simcoe, Ontario, Canada and North Carolina had sorption properties for chlorothalonil that were nearly the same. For labile surface sorption kinetics, the Simcoe soil gave a pseudo first-order rate constant of kS1 = (7.4 +/- 0.7) x 10(-2) days-1. At equilibrium, the labile surface sorption capacity theta c of Simcoe soil for chlorothalonil was 23.8 x 10(-6) (mol/g). The sorption properties of the 4-hydroxy derivative of chlorothalonil were different in two important respects. They were larger by an order of magnitude, and they were substantially different for the two soils. Sorption by the Simcoe soil was too fast for kinetics measurements by the on-line HPLC micro extraction method, but for the North Carolina soil kS1 = (1.15 +/- 0.01) days-1 was recorded. For the Simcoe and North Carolina soils, respectively, theta c > 200 (mumol/g) and theta c approximately 113 (mumol/g). Two conclusions can be drawn. First, the replacement of the Cl by OH on the 4 position of chlorothalonil makes the sorption effects much greater. Second, the stronger interactions are associated with a greater sensitivity to small differences in the chemical compositions of the soils. Subtle soil properties causing significant effects might include small amounts and physical structures of organic matter and metal oxides. This implies that, for predictive computer models, mechanism parameters will have to be correlated in two dimensions: chemical structure, and the composition and amounts of chemical materials in soils. PMID- 11414049 TI - Study of Au/Au(3+)-TiO2 photocatalysts toward visible photooxidation for water and wastewater treatment. AB - With an attempt to extend light absorption of TiO2-based photocatalyst toward the visible light range and eliminate the rapid recombination of excited electrons/holes during photoreaction, a new type of photocatalysts (Au/Au(3+) TiO2) powder was prepared by a photoreduction/sol-gel process. The crystal phase composition, surface structure, and light absorption of the new photocatalysts were comprehensively examined by X-ray differential detection (XRD), UV-visible absorption spectra, X-ray photoelectron emission spectroscopy (XPS), and photoluminescence (PL) spectra. The photooxidation efficiencies of the photocatalysts were also evaluated in the photodegradation of methylene blue (MB) in aqueous solutions under visible light irradiation from a high-pressure sodium lamp (lambda > 400 nm). The results of PL analyses in this study indicated that the gold/gold ion-doping on the surface of TiO2 could eliminate the electron/holes recombination and also increase the light absorption in the visible range. The analytical results of UV-visible diffuse reflection spectra (DRS) and optical absorption spectra indicated that a new energy level below 3.2 eV generated in the Au/Au(3+)-TiO2 promoted the optical absorption in the visible region and made it possible to be excited by visible light (E < 3.2 eV). The experiment demonstrated that the photooxidation efficiency of MB using the Au/Au(3+)-TiO2 powder were significantly higher than that using conventional TiO2 powder and an optimum molar content of gold doping/deposition in the TiO2 was 0.5%. The development of such photocatalysts may be considered a breakthrough in large-scale utilization of solar energy to address environmental needs. PMID- 11414050 TI - Hybrid organic/inorganic reverse osmosis (RO) membrane for bactericidal anti fouling. 1. Preparation and characterization of TiO2 nanoparticle self-assembled aromatic polyamide thin-film-composite (TFC) membrane. AB - Hybrid organic/inorganic reverse osmosis (RO) membranes composed of aromatic polyamide thin films underneath titanium dioxide (TiO2) nanosized particles have been fabricated by a self-assembly process, aiming at breakthrough of biofouling problems. First, positively charged particles of the colloidal TiO2 were synthesized by a sol-gel process, and the diameter of the resulting particles in acidic aqueous solution was estimated to be approximately 2 nm by analyzing the UV-visible absorption characteristics with a quantum mechanical model developed by Brus. Transmission electron microscopy (TEM) further confirmed the formation of the quantum-sized TiO2 particles (approximately 10 nm or less). The TiO2 particles appeared to exist in the crystallographic form of anatase as observed with the X-ray diffraction (XRD) pattern in comparison with those of commercial 100% rutile and commercial 70:30% anatase-to-rutile mixture. The hybrid thin-film composite (TFC) aromatic polyamide membranes were prepared by self-assembly of the TiO2 nanoparticles on the polymer chains with COOH groups along the surface. They showed improved RO performance in which the water flux even increased, though slightly. Field-emission scanning electron microscopy (FESEM) exhibited the TiO2 nanoparticles well adsorbed onto the surface. X-ray photoelectron spectroscopy (XPS) demonstrated quantitatively that a considerable amount of the adsorbed particles were tightly self-assembled at the expense of the initial loss of those that were loosely bound, and became stabilized even after exposure to the various washing and harsh RO operating conditions. The antibacterial fouling potential of the TiO2 hybrid membrane was examined and verified by measuring the viable numbers and determining the survival ratios of the Escherichia coli (E. coli) as a model bacterium, both with and without UV light illumination. The photocatalytic bactericidal efficiency was remarkably higher for the TiO2 hybrid membrane under UV illumination, compared to that of the same membrane in darkness, as well as those for the neat membranes under either light condition. PMID- 11414051 TI - Removal of naphthalene, phenanthrene, and pyrene by sorbents from hot gas. AB - It is the first time that the removal of polycyclic aromatic hydrocarbons (PAH) containing different aromatic rings number [naphthalene (Np), phenanthrene (Phe), and pyrene (Py)] from combustion hot gas has been carried out. The aim was to relate the sorbents textural characteristics with the adsorption capacity of these 2-4-ring PAH at the conditions emitted at energy generation. The sorbents textural parameters [total micropore volume (VN2), narrow micropore volume (VCO2), mesopore volume (VBJH), and the free active sites] were analyzed trying to correlate them with their Np, Phe, and Py adsorption capacities. To get this aim, single and multiple linear regressions (MLR) were applied to the three PAH. A principal component analysis was performed to generate new and uncorrelated variables. It enabled us to show that the relations between the textural parameters were analyzed using a principal components regression (PCR). The PCR analysis had a good statistical quality, but neither did it allow differentiating free active site types nor did VN2 and VCO2. The correlations were thus set up applying a MLR to the original variables. The regression statistical quality was similar to the PCR analysis, and it could give an easier explanation of the parameters that affected the adsorption. In Np adsorption, the 87% data variance was explained, and the adsorption was positively correlated to VCO2 and the micropore mean diameter (I.). In the Phe regression there was 98% variance explained, and its adsorption was positively correlated to the VN2 and the micropore distribution, n. Finally, in the Py adsorption, the 96% data variance was explained, and this adsorption was positively correlated to VN2 and VBJH. These dependencies were according to the molecular parameters of these compounds (molecular diameter and volatility) because the higher the number of aromatic rings of the PAH, the more favored the adsorbate-adsorbate interactions. Besides, the higher the mean diameter micropores, the lower the diffusional problems showed by Np, Phe, and Py. PMID- 11414052 TI - Will drilling for oil disrupt the Arctic National Wildlife Refuge? PMID- 11414053 TI - Making TMDLs work. PMID- 11414054 TI - TMDL at the crossroads. PMID- 11414055 TI - [Assessment of pure isolated mitral valve insufficiency by Doppler echocardiography]. AB - Taking 33 patients having pure MI as a material, the authors find a correlation between regurgitation fraction obtained by calculation of outputs estimated by Touch's method and angiographic values. There is a statistically significant differences (P < 0.001) between regurgitation fraction of grade I to II and grade II to III MI. The ratio mitral integral time velocity (ITV) to Aortic (ITV) is an important semi-quantitative assessment of pure MI. In fact, a ratio > 1.3 identify important degree of MI with 82% sensitivity and 93% specificity. The authors estimate that there is a correlation between the ratio of regurgitant jet surface to left atrial surface found in TEE and their degree of MI in angiography with a significant difference (P < 0.001) between the ratio of grade I to II and grade II to III MI in angiography. A ratio higher than 40% allow to identify grade III MI at minimum. PMID- 11414056 TI - [Submandibular lithiasis. Report of 48 cases]. AB - This retrospective study concerned 48 patients treated surgically for sub mandibular lithiasis. Results of medical imaging (plain films, sialography, ultrasonography) are confronted with operative findings. The surgical indications are analysed according to lithiasis localisation. PMID- 11414057 TI - [Medical termination of pregnancy. Indications, techniques and complications. Report of 55 cases]. AB - We tried through this study to reassemble the cases of medical termination of pregnancy for foetal or maternal anomalies and analyse the epidemiological characteristics of our population and the procedures of induction of labour. We report a retrospective study about 55 cases of medical termination of pregnancy during 20 months indexed in service "C" of the centre of maternity and neonalogy in collaboration with the service of fetopathology. 12.73% of our patients were older than 38 years. The incidence of medical termination of pregnancy during the same period was about 1.90 cases/100 deliveries. Maternal indications was reported in 34.55% of cases and neurological malformations dominate the foetal anomalies (47.22%). Extra-amniotic saline infusion was used in 52.72% of cases and was responsible of all the complications observed (16.36%). PMID- 11414058 TI - [Pancreatic pseudocysts. Report of 13 cases at the Central University Hospital at Lome-Tokoin]. AB - A prospective study done over a period of ten years in which a 13 dossiers of patients with pseudopancreatic cysts due to complication of acute pancreatitis, chronic pancreatitis and one pancreatic traumatism were collected. They were 12 male, one female with age incidence between 14 years and 60 years with an average age of 37 years. Twelve cases were confirmed by abdominal echography one case was diagnosed by indirect signs of oesophago-gastroduodenal transit. There were 4 cases of pseudocysts, 8 cases of retention cysts and one case post traumatic pseudocyst. Eleven patients were operated, 7 cases by cystojejunostomies on Y shape technique, one case with external drainage, and one case by pancreatectomy of the tail. PMID- 11414059 TI - [Severe chronic hypoxemia in cirrhosis]. AB - In the course of cirrhosis, severe chronic hypoxemia (< 60 mmHg) is exceptional, it is the most often in contact with intrapulmonary vascular anomalies and necessitates complex investigations. Authors reported a case of 16-years old patient, breach of cirrhosis with underdevelopment and that presents a severe chronic respiratory insufficiency not improved by the addition of oxygen. The different practiced explorations are in favor of an hepatopulmonary syndrome secondary probably to intrapulmonary shunting. PMID- 11414060 TI - [Cardiac sarcoidosis revealed by ventricular tachycardia]. AB - Cardiac sarcoidosis is a multisystem granulomatous disease of unknown etiology, that is more seen in younger adults. The prognosis of myocardial sarcoidosis is poor and depends on arrhythmia, conduction disorders. We report the case of 35 years old woman with cardiac sarcoidosis and pulmonary involvement who presented acutely with sustained ventricular tachycardia well tolerated. She made a good recovery after steroid therapy and administration of antiarrhythmic agent. PMID- 11414061 TI - [Right paraduodenal hernia: a case report]. AB - Paraduodenal hernia is a rare etiology of intestinal obstruction. Its preoperative diagnosis can only be made by a well-performed upper gastrointestinal series. We report a case of a 19 years old man with abdominal pains and recurrent Koenig's syndromes. The upper gastrointestinal series and CT scan showed a right paraduodenal hernia. we reviewed the anatomy. physiopathology, symptoms and radiographic criteria for the diagnosis of the paraduodenal hernia. PMID- 11414062 TI - Mycobacterium recombinant vaccines. AB - Intracellular diseases are difficult to treat and constitute a major problem for modern medicine. In this type of diseases, a TH-1 immune response favors protection, while a TH-2 response is detrimental to the host. Current vaccines are using antigens to initiate an immune response regardless of its nature and its mechanism. New vaccines are designed to combine selected antigens with potent adjuvants to stimulate the appropriate pathway of the immune system and deliver a lasting protective immunity. The Mycobacterium recombinant vaccine system for treatment of intracellular diseases utilizes antigen delivery systems in the form of non pathogenic Mycobacterium strains, genetic transfer systems in the form of cloning and expression vectors, and related technologies to provide products containing non toxic immuno-regulating Mycobacterium adjuvants, non toxic immuno stimulating exogenous antigens specific for a variety of diseases, and non toxic amounts of cytokines that boost the TH-1 pathway. The cloning and expression Mycobacterium vectors include both pAL5000-based extra-chromosomal and D29-based integrative vectors. PMID- 11414063 TI - [Non-pharmacologic treatment of atrial fibrillation]. AB - Curative non pharmacological treatment of AF includes MAZA anti arrhythmic surgery and ist different variants which are reserved to AF associated with mitral valve disease. Radiofrequency (RF) ablation of AF creating lesions mimicking the MAZE procedure is another curative treatment but its numerous complications lead to put it in stand-by. Focal AF radiofrequency ablation is the only curative method that has encouraging results. The development of the internal cardioversion led to the development of the implantable atrial defibrillator. In spite of the low energy used, the internal shocks are often painful and not well tolerated conducting to stop the use of these devices. Prophylactic atrial pacing among patients with interatrial conduction delay is well established by dual site pacing pioneers. After several attempts to restore sinus rhythm and the fail of drug therapy to reduce heart rate, the latest solution to prevent tachycardiomyopathy is the atrio-ventricular (AV) node modulation or ablation using RF energy. PMID- 11414064 TI - [Detection and significance of cryoglobulins]. AB - Cryoglobulins are immunoglobulins which reversibly precipitate at cold temperatures. They are usually accompanied with polymorphic clinical manifestations and biological abnormalities. Cryoglobulins are associated with various clinical disorders which include autoimmune diseases, lymphoproliferative and infectious disorders. Because of their low concentration, detection and quantitation of cryoglobulins need very rigorous conditions when collecting blood from patients. PMID- 11414065 TI - [Health indicators in Tunisia, trends in regional disparities over the last thirty years]. AB - The measure of health indicators on a regional-scale seems more interesting then the measure on a national-scale. It allows us to study differences between regions and their evolution. In order to know the time trend's disparities between regions, we have studied seven indicators, concerning different types of the World Health Organisation's classification, using data from yearly statistics and results of decennial census. Regional disparities are evaluated by a ratio between the highest regional value and the lowest regional value. This study showed us a global trend to the decline of regional disparities about indicators that we have studied. The improvement of health care resources indicators and socioeconomic's indicators, in the process of time, is associated with the improvement of health status indicators. The study of regional disparities makes possible to target actions for health in order to promote health's equity between regions. PMID- 11414066 TI - [Study of the effect of converting enzyme inhibitors on microalbuminuria in diabetics]. AB - The authors report the results of a randomised trial using a converting enzyme inhibitor in 40 microalbuminuric diabetic subjects during 18 months. In the treated group, we observed a reduction of albuminuria from 57.4 mg/24 hours to 35.4 mg/24 hours at the end of the follow up, in contrast with a non significant progression in the group who didn't receive this medication. No significant modification in the clinical and biological parameters was observed during the follow up. PMID- 11414067 TI - Reproductive technologies and human experimentation: a Jewish perspective. PMID- 11414068 TI - Privacy legislation and health care. PMID- 11414069 TI - Internet pharmacy: options for Canadian regulation. PMID- 11414070 TI - Total hip and knee replacement surgeries in Alberta utilization and associated outcomes. AB - OBJECTIVE: Between 1994/95 and 1996/7, the healthcare system in Alberta underwent a radical transformation that saw the regionalization of service delivery and a reduction in overall healthcare funding. The aim of this study is to examine trends in utilization and associated post-operative complications and readmissions for two procedures, total hip arthroplasty (THA) and total knee arthroplasty (TKA) during this period of restructuring. METHODS: This is a six year retrospective analysis using hospital separation and healthcare insurance plan registry databases. During the study period, we looked at trends in inpatient procedure volumes, average lengths of stay, post-operative complications during the index hospital stay and readmissions within one year. RESULTS: Over a six-year period, the age-sex standardized provincial rate of THAs increased by 18% while the rate of TKAs increased by 73%. Average lengths of stay decreased for both procedures by over 50%. The rate of procedures with at least one post-operative complication within the index episode of care decreased over the study period by 19% for THA and 21% for TKA. Readmissions due to complications of surgery increased overall by 32% for THA and 2% for TKA. Regional and sex differences were observed throughout the period. CONCLUSIONS: While the overall rate of hospitalizations has decreased in Alberta, the rates of THA and TKA procedures have increased. The lengths of stay following these procedures have, at the same time, reduced dramatically. Adverse outcomes as measured by complication rates associated with these procedures have decreased, though readmissions due to complications after surgery have increased. PMID- 11414071 TI - Hypertensive patients and their general practitioners. AB - This study examined the relative contribution of hypertensive patients and their physicians in selecting total annual physician visit frequencies and made regional comparisons between two Canadian cities. We found that the frequency of physician visits was primarily influenced by physician referrals and physician practice patterns, which accounted for about 80 percent of the total explainable variance in physician visits. The relative contribution of other available patient and physician characteristics in determining visit frequency was rather small. PMID- 11414072 TI - Healthcare quality in an international setting. AB - This report discusses the process used by the Healthcare Quality Certification Board to internationalize its certification program. With some modifications this methodology can be utilized to define the role of any healthcare professional and to legitimize, create or restructure a certification program that assesses competency in a particular healthcare field. This process and the importance of quality management are related to today's cataclysmic healthcare environment which challenges efforts to confront cost and access. PMID- 11414073 TI - Strides in care management: the development of a framework to facilitate quality outcomes. AB - Care Management has been in existence for years. The thrust of the role is to monitor, evaluate and enhance resource utilization while maintaining quality of care. The pivotal assumption being through enhanced patient flow, and coordinated services with appropriate resource utilization, better patient outcomes are achieved. During times of fiscal restraint, quality may become a secondary concern. This paper presents a framework to assist Care Managers, and administrators, in achieving desired outcomes that embrace fiscal accountability and quality patient care. Donabedian's model and Systems theory are the foundations for this framework. PMID- 11414074 TI - Tuberculosis in the eastern European countries and the former USSR. AB - This article provides a situational analysis of the increasing trends in the tuberculosis case notification in Eastern Europe and the former USSR. In the Russian Federation, one main reason for the spread of tuberculosis is the insufficient funding of TB control measures which were very costly in the past and which cannot be sustained due to the current economic problems. The author also describes his own experience as technical advisor to a TB control program in western Siberia. PMID- 11414075 TI - Exploring a chronic care model in a regional healthcare context. AB - The episodic acute care model of service delivery that consumes the majority of healthcare funding is inadequate to meet the needs of a society experiencing an increasing burden of chronic illness. This article describes, in a regional Canadian healthcare context, the application and preliminary evaluation of a model of chronic care delivery developed in a managed care environment in the United States. The strategic goals of this model are to empower clients by providing a redesigned service delivery environment that supports self management; support care providers by developing clinical information systems and decision supports; and align community resources, policies and organization of care to support an informed, activated client and a proactive practice team. PMID- 11414076 TI - Changing characteristics of carotid endarterectomy. AB - We have noted significant differences in terms of our preoperative work-up, length of stay, morbidity, and mortality of patients undergoing carotid endarterectomy (CEA) from findings reported in large published randomized clinical trials. To further investigate these differences, we have reviewed our recent experience. CEA has proved to be the most effective approach to avert stokes caused by significant atherosclerotic disease of the carotid bifurcation. Between January 1, 1996 and December 31, 1998, 552 patients underwent CEA at our institution. Forty percent were performed in symptomatic patients with stenotic lesions > 60% in diameter by duplex ultrasonography. The remainder were performed for asymptomatic lesions > 60% in diameter. No patient underwent contrast angiography. Fifty-two percent of the patients were males. The mean age was 74 +/ 8 years old. General anesthesia was used in 97% of the cases and regional block, in 3%. All patients underwent routine postoperative measurement of serum creatinine phosphokinase (CPK) isoenzymes. Patients were discharged when deemed clinically stable. The patients' follow-up visits at 1 week and at 3-5 months after the procedure (mean, 3.4 months) included a neurological exam and duplex exam. Patient results suggest that CEAs can be performed in the modern era without contrast arteriography. Most patients can be discharged on the first postoperative day. In addition, previously acceptable rates of postoperative morbidity and mortality should perhaps be revised to meet current standards. Contrary to the previous concept that most postoperative strokes are due to embolic phenomena, hyperperfusion syndrome played an increasingly important role in this review. PMID- 11414077 TI - Long-term follow-up of saphenous vein, internal jugular vein, and knitted Dacron patches for carotid artery endarterectomy. AB - To determine whether choice of material used for patch closure following carotid artery endarterectomy (CAE) influences rates of early or late restenosis, stroke, and death, 274 consecutive CAEs were retrospectively reviewed. Saphenous vein (SV) was used in 159 (58.0%) procedures; everted, double-thickness jugular vein (JV) was used in 25 (9.1%); and knitted Dacron (KD) was used in 90 (32.9%). Primary closure was not used in this series. There were four perioperative strokes: two (1.3%) in SV, one (4%) in JV, and one (1.1%) in KD (NS). Follow-up was obtained on 263 (96%) operated arteries (mean 41.5 months). Duplex scan results were available for 236 (89.7%) of these arteries (mean follow-up time 33.7 months). There were three (2%) late strokes in SV and two (2.2%) in KD (NS). In long-term follow-up, one patient (0.7%) in SV and two (2.4%) in KD developed > 80% stenosis (NS). One patient (0.7%) in SV, one (5.3%) in JV, and one (1.2%) in KD had total occlusion of the operated vessel (NS). Three procedures (2.2%) in SV, 1 (5.3%) in JV, and 7 (8.5%) in KD demonstrated moderate stenosis (50-79%) (NS). Three-year follow-up shows that choice of patch material does not affect early or late stroke rate, stroke-related death rate, rate of high-grade (> 80%) restenosis, or rate of total occlusion. There is a higher incidence of moderate stenosis in KD. Although our results and a review of the literature do not indicate that these patients are at increased risk for symptoms or progression of stenosis, they should be followed by duplex scanning to ensure that this is the case. PMID- 11414078 TI - A strategy to improve quality of outpatient education for residents and medical students on a vascular surgery service. AB - In the current era of same-day admissions and decreased length of hospitalization for major vascular operations, education of residents and medical students in the outpatient setting has become an increasingly challenging problem. We developed a strategy to provide improved outpatient experience for advanced vascular surgery residents (VSRs), general surgery residents (GSRs), and medical students (MSs) on a vascular surgery service. A survey of program directors and VSRs at accredited vascular surgery fellowships in the United States and Canada was undertaken to determine the manner in which outpatient education was accomplished and the amount of time devoted to pre- and postoperative patient evaluation. The survey revealed that VSRs, GSRs, and MSs spent 1 or more days in clinic at approximately half of accredited vascular surgery programs. There were no appreciable differences in time spent in clinic for different levels of trainees. VSRs preoperatively evaluated at least half of their patients at approximately two thirds of the programs, while at the other third of the programs they preoperatively evaluated one-quarter or less of their patients before surgery. GSRs preoperatively evaluated at least half of their patients at 50% of programs, according to program directors, and at 69% of programs, according to VSRs. Because of recent changes in health care delivery and the effect of these changes on resident training, outpatient education of VSRs, GSRs, and MSs will require increased attention on the part of vascular surgeons involved with their education. increasing time demands and decreasing reimbursements for vascular surgeons as a result of managed health care may make this goal increasingly difficult to accomplish. Improved strategies and extra efforts are necessary to ensure that trainees obtain sufficient experience to evaluate vascular patients pre- and postoperatively and maintain high quality of care for vascular surgery patients. PMID- 11414079 TI - Current training and practice of endovascular surgery: a survey. AB - Endovascular surgery (EVS) has become of great interest to the vascular community. However, little data exist about the current status of training and utilization of EVS, thus prompting this survey. During a national vascular meeting, all participants (n = 132) filled out a 2-page questionnaire. Respondents (mean age 39 years, 87% male) were 68 vascular surgeons in practice (52%), and 64 fellows (48%), representing a significant fraction of trainees in North America. Practice location included university hospital (39%), private hospital/clinic (52%), and government hospital (8%). According to respondents, in their respective communities, most of the EVS was performed by radiologists (66%), followed by vascular surgeons (19%), cardiologists (13%), and vascular medicine physicians (2%). A majority of respondents (75%) currently perform EVS; surgeons in practice < 3 years had the highest rate (90%). Utilization rates among the nine interventions surveyed ranged from angiography (72%) and angioplasty (65%) to intravascular ultrasound (IVUS) (21%) and atherectomy (12%). Procedure totals showed that approximately 20% of fellows performed > 100 angiograms and 10% performed > 25 endovascular grafts/year, whereas nonfellows performed few of the latter. Most of the EVS (72%) was performed in the operating room with portable imaging equipment and EVS accounted for 14% of all vascular procedures. Most respondents (86%) believed that EVS would become a major component of vascular surgery and comprise 30% of their future practice. Seventy six percent thought their training was insufficient and 85% said they would devote 3 months or less for further endovascular training. In conclusion, a vast majority of vascular trainees and surgeons are performing EVS, however, individual caseloads vary greatly. The belief that endovascular surgery will play an increasing role in vascular surgery practice is strong and interest in further training of short duration is widespread. Broader-based data collection and longitudinal studies on this issue are warranted. PMID- 11414080 TI - Operative results and death predictors for nonruptured abdominal aortic aneurysms in the elderly. AB - Elective repair of abdominal aortic aneurysms (AAA) in the elderly has conventionally been associated with higher mortality rates than in younger patients, although some authors do not agree with this view. The aim of this study is to look into current results of surgical treatment in elderly patients with nonruptured AAA and to define those factors associated with an increased mortality. Over a period of 2 years (1995-1996), 183 patients aged 75 or older were operated on for nonruptured AAA in 21 hospitals. Their medical records were reviewed and those factors that could be related to death were analyzed. Those variables with statistical significance through univariate analysis were entered into a multivariate logistic regression model to predict mortality. The results showed that surgery for nonruptured AAA in the elderly has a greater mortality rate than that in younger patients, but it is not so great as to deny surgical correction to prevent rupture in selected cases, especially if they are unsuitable for endovascular repair. These patients must first undergo thorough cardiac and renal function evaluation. Selective screening must be considered, so earlier detection and treatment in younger patients can be performed. PMID- 11414081 TI - Embolization of hypogastric artery aneurysm: 17 cases. AB - Surgical management of hypogastric artery aneurysm is associated with high morbidity due to hemorrhage and ischemia. Occlusion by embolization is an attractive alternative treatment. Between 1991 and 1995, we used Gianturco coils to embolize 17 hypogastric aneurysms in 14 patients. All patients were men with a mean age of 77 years. Ten patients had previously undergone aortic repair. Complete occlusion of the aneurysm was achieved in 16 cases but placement of an iliac stent was required in 1 case. Embolization failed in one case involving rupture of a large aneurysm. No complications were observed. Moderate buttock claudication was noted after bilateral embolization in three cases. Embolization of hypogastric artery aneurysm using coils resolves the long-term problems associated with surgical ligation. Extensive aneurysm of the origin to the bifurcation is the main indication for nonresective treatment but embolization can also be a useful alternative to open surgery for high-risk patients. Availability of stent grafts may extend the indication for endovascular treatment. PMID- 11414082 TI - Aneurysms of the internal iliac artery: management strategy. AB - With the widespread use of CT scans, detection and treatment of internal iliac artery aneurysms (IIA) have become more frequent. In the last few years, endovascular repair has been added to the therapeutic arsenal. We reviewed the records of 38 patients treated for 44 IIA between 1987 and 1997 to assess immediate and long-term outcome using various therapeutic methods. Aneurysms were divided into three groups according to the circumstances of treatment. Group I included 25 IIA treated at the same time as abdominal aortic aneurysm (AAA). The morbidity/mortality rate in this group was comparable to that in patients who underwent isolated AAA repair. Group II included 14 IIA treated during follow-up of AAA repair. Most complications in this group were intraoperative. Group III included five isolated IIA not associated with AAA repair. Complications were similar to those in group I. On the basis of this retrospective analysis, we propose a management strategy in which open surgery, endovascular repair, or both are used, depending on the circumstances of treatment. PMID- 11414083 TI - Gut mucosal injury is attenuated by recombinant bactericidal/permeability increasing protein in hind limb ischemia-reperfusion injury. AB - Lower limb ischemia-reperfusion injury (IRI) is associated with increased gut permeability to endotoxin, which not only directly damages enterocytes but also stimulates a systemic inflammatory response syndrome (SIRS), compounding gut injury. Recombinant bactericidal/permeability-increasing protein (rBPI21) is a novel anti-endotoxin therapy with proven benefit in sepsis. Its potential role in modulating remote gut injury in hind limb IRI was studied. Male Wistar rats were chosen for a prospective randomized control trial (n = 10 per group). The control group and two groups undergoing 3 hr bilateral hind limb ischemia with 2 hr reperfusion (I/R) were randomized to receive intravenously either control protein thaumatin at 2 mg/kg or rBPI21 at 2 mg/kg, respectively. Quantitative morphometric assessment of the small bowel was used as a measure of gut injury and, using an ex vivo everted gut sac model, translocation of 14C-labeled polyethylene glycol (PEG) was used as a measure of gut permeability. Our results indicate that hind limb IRI is associated with remote gut mucosal injury and increased permeability to macromolecules. rBPI21 anti-endotoxin therapy modulates remote gut injury associated with lower limb IRI in this model. PMID- 11414084 TI - Alcohol withdrawal after open aortic surgery. AB - This study was designed to test the hypothesis that unexpected alcohol withdrawal like syndrome (AWLS) is more common following aortic, but not other, vascular or nonvascular procedures. All patients undergoing open aortic surgery at our institution in 1997 who survived at least 48 hr were identified, as were those undergoing carotid endarterectomy, infrainguinal bypass, and total colectomy. AWLS was defined as prolonged confusion or agitation and response to conventional treatment for withdrawal, providing that all other sources had been ruled out or a significant history was present. Our results show that, for unknown reasons, AWLS is more common after aortic surgery than after other vascular and high stress, nonaortic intraabdominal procedures at our institution, and is associated with increased length of stay and morbidity. Because prophylaxis may improve outcome, better efforts to identify patients at risk are required. PMID- 11414085 TI - Arcuate-legged nonpenetrating vascular closure staples (VCS): early experience. AB - Vascular closure staples (VCS) provide a novel technique for fashioning vascular anastomoses, allowing a single operator to perform suture-less anastomoses. They may be used primarily or in an adjuvant role. When VCS are compared to a running suture, advantages include the avoidance of intimal damage, platelet aggregation and intimal hyperplasia at the anastomotic suture line, and a shorter time taken to complete the anastomosis. We report our early experience using VCS in an array of vascular anastomoses and conclude that VCS are a useful addition to the vascular surgeon's armamentarium. They help to decrease the time taken to construct an anastomosis, and are particularly useful in an adjuvant setting, complementing conventionally placed sutures. PMID- 11414086 TI - Mechanical determinants of graft kinking. AB - Vascular grafts can be twisted inadvertently during implantation. If twisted excessively, they may kink and obstruct flow. In this study, in vitro experiments were performed to identify the mechanical factors that determine graft kinking. These included graft material, graft length, graft diameter, graft wall thickness, perfusion pressure, and flow rate. Six-millimeter-diameter saphenous veins were obtained from humans at autopsy. Six-millimeter standard-wall and 6-mm thin-wall polytetrafluoroethylene (PTFE) grafts also were obtained. Both fixed length and stretchable PTFE grafts were examined. Grafts 15, 30, 50, and 70 cm in length were evaluated. Finally, PTFE grafts 4, 6, 8, 10, 12, and 14 mm in diameter were studied to determine the effect of diameter. The vessels were mounted horizontally in vitro and were perfused with saline at 50, 100, or 150 mmHg pressure at low (49 mL/min), medium (105 mL/min), and high (239 mL/min) flow rates. Each graft was twisted 90 degrees, then subjected to perfusion for 15 sec. Pressure and flow were interrupted, and an additional 90 degrees twist was imposed for another 15 sec. This sequence was repeated until a visible kink developed. We conclude from our results that, when constructing a bypass, particular care should be taken with vein, short grafts, thin-wall grafts, and large-diameter grafts, as these are especially susceptible to kinking. PMID- 11414087 TI - Effectiveness of endothelial cell seeding on patency of damaged vascular surfaces in a canine model. AB - The effects of endothelial cell seeding, which is assumed to be an effective technique to improve patency rates of denuded vascular surfaces, were investigated in an experimental model. In this study, after anesthetic induction, jugular veins of 16 dogs were harvested bilaterally. Endothelial cells were extracted enzymatically by collagenase from these veins and were passaged into a culture medium until they grew to a reasonable number. After 3 weeks, dogs were anesthetized again in a similar fashion and bilateral femoral veins were exposed and experimental intimal denudation was performed. Subsequently, one femoral artery was injected with cell solution and the other with saline solution as a control. Two weeks after the injections, arteriographic studies of femoral arteries were performed and arterial specimens were taken for histological evaluation. Our results suggest that endothelial seeding might improve the patency rate in elective but urgent cases in which endarterectomy, percutaneous transluminal angioplasty, or similar vascular procedures are considered. PMID- 11414088 TI - Wall stress studies of abdominal aortic aneurysm in a clinical model. AB - To estimate when an abdominal aortic aneurysm (AAA) may rupture, it is necessary to understand the forces responsible for this event. We investigated the wall stresses in an AAA in a clinical model. Using CT scans of the AAA, the diameter and wall thickness were measured and the model of the aneurysm was created. The wall stresses were determined using a finite element analysis in which the aorta was considered isotropic with linear material properties and was loaded with a pressure of 120 mmHg. The AAA was eccentric with a length of 10.5 cm, a diameter of 2.5 to 5.9 cm, and a wall thickness of 1.0 to 2.0 mm. The aneurysm had specific areas of high stress. On the inner surface the highest stress was 0.4 N/mm2 and occurred along two circumferentially oriented belts--one at the bulb and the other just below. The stress was longitudinal at the anterior region of the bulb and circumferential elsewhere, suggesting that a rupture caused by this stress will result in a circumferential tear at the anterior portion of the bulb and a longitudinal tear elsewhere. In the mid-surface the highest stress was 0.37 N/mm2 and occurred at two locations: the posterior region of the bulb and anteriorly just below. The stress was circumferential, suggesting that the rupture caused by this stress will produce a longitudinal tear. The location and orientation of the maximum stress were influenced more by the tethering force than by the wall thickness, luminal pressure, or wall stiffness. In conclusion, the rupture of an AAA is most likely to occur on the inner surface at the bulb. Such analytical approaches could lead to a better understanding of the aneurysm rupture and may be instrumental in planning surgical interventions. PMID- 11414089 TI - Adenoviral-mediated gene transfer in human and animal vein grafts using clinically relevant exposure times, pressures, and viral concentrations. AB - This study examined the efficiency of adenoviral-mediated gene transfer in experimental vein grafts and cultured human saphenous vein under physiologic conditions using clinically relevant exposure times, pressures, and viral concentrations. The external jugular veins of 25 male New Zealand White rabbits were exposed to 0.5 mL of replication-deficient adenovirus vectors encoding beta galactosidase (AdlacZ), control adenovirus (AdBg/II), or vehicle at pressures ranging from 0 to 120 mmHg for 10 min. Veins were excised and grafted into the carotid circulation. After 5 days, the vessels were reexposed, excised, and stained with X-gal chromagen for beta-galactosidase (beta-gal) activity. Gene transfer was also performed in 13 segments of human saphenous vein discarded at the time of bypass grafting. The veins were cultured for 0-21 days and assayed for beta-gal activity as above. Rabbit vein grafts exposed to high-pressure AdlacZ transfection showed significant transgene expression in 100% of grafts (39 +/- 2% positive cells/hpf) while only 60% of those transfected at low pressure expressed beta-gal (9 +/- 3% positive cells/hpf). All human veins exposed to AdlacZ expressed beta-gal to a variable degree (range 10-50% positive cells/hpf). No control grafts or veins expressed the transgene. Efficient adenoviral-mediated gene transfer in experimental vein grafts and human saphenous vein segments can be achieved using clinically feasible parameters of exposure time, pressure, and viral concentration. PMID- 11414090 TI - Contemporary analysis of outcomes following lower extremity bypass in patients with end-stage renal disease. AB - This study was designed to compare outcomes following infrainguinal bypass between patients with end-stage renal disease (ESRD) and patients with normal renal function (NRF). Sixty-three patients with ESRD undergoing 78 infrainguinal bypasses from 1990 to 1999 were compared with a concurrent group of 132 age-, race-, and gender-matched patients with NRF undergoing 148 bypasses. Limb salvage and survival were calculated using Kaplan-Meier analysis. Markov decision analysis was used to calculate expected quality-adjusted life years (QALY) with intervention. Mean follow-up was 25 months (range 1-116). The results of our study show that infrainguinal bypass in patients with ESRD and tissue necrosis appears to provide a measurable, but marginal, degree of improvement in quality of life. PMID- 11414091 TI - Is the preferential use of ePTFE grafts in femorofemoral bypass justified? AB - The choice of prosthetic graft material for cross-femoral bypass has been evolving in the past two decades. Expanded polytetrafluoroethylene (ePTFE) has become our preferred graft material since 1995. However, few studies have looked into the optimal graft material in this procedure. Justification for the preferential use of ePTFE graft in lower limb revascularization remains unknown. The aim of the present study was to compare the long-term outcomes of Dacron and ePTFE grafts in femorofemoral bypass. The records of 61 consecutive patients who underwent femorofemoral bypass at the University of Hong Kong Medical Center from 1981 to 1998 were retrospectively reviewed. Dacron grafts were used in 27 patients and 34 patients had ePTFE grafts. The demographic features, patency, and limb salvage rates of the two groups of patients were compared. The 3-year primary patency rates of Dacron and ePTFE grafts were 85% (SE = 9.5%) and 66% (SE = 14.5%), respectively. The difference was not statistically significant. The limb salvage rates of Dacron and ePTFE grafts were 91% and 83% at 3 years, respectively (p = 0.27). The long-term outcomes of Dacron and ePTFE grafts in femorofemoral bypass were equivalent. The preferential use of ePTFE graft in femorofemoral bypass is not evidence based. Selection of an appropriate prosthetic graft for femorofemoral bypass should be based on the cost and its handling characteristics. PMID- 11414092 TI - Regulation of VEGF in diabetic patients with critical limb ischemia. AB - Diabetic patients are at a 10- to 20-fold increased risk for the development of critical limb ischemia. Vascular endothelial growth factor (VEGF) is critical for the development of collateral blood vessels, which can effectively bypass peripheral arterial occlusions. We therefore set out to determine if the regulation of VEGF in patients with peripheral vascular disease differs in diabetic and nondiabetic patients. Diabetic and nondiabetic patients with peripheral vascular disease were divided into those with or without critical limb ischemia as defined by clinical criteria (rest pain, nonhealing ulcer). Monocytes from peripheral blood were isolated from all patients and the hypoxic induction of VEGF was determined in vitro. In patients without diabetes, we found that there was no significant difference in the hypoxic induction of VEGF between patients with or without critical limb ischemia. However, in diabetic patients we found that patients with critical limb ischemia produced significantly more VEGF than patients without critical limb ischemia (6.3 +/- 1.3 vs. 2.1 +/- 0.3, p < 0.015). We conclude that diabetic patients with critical limb ischemia do not have an impairment in the ability to produce VEGF with hypoxia. Contrary to current dogma, treatment paradigms directed at increasing VEGF production in the diabetic patient with critical limb ischemia might not be beneficial. PMID- 11414093 TI - Arterial mycotic aneurysm rupture following kidney-pancreatic transplantation with exocrine pancreatic drainage into the bladder: an unusual observation. AB - In this case report, we describe two patients who received a kidney-pancreatic transplant through technique of exocrine pancreatic drainage into the bladder, and who subsequently developed arterial mycotic aneurysms at the site of the arterial anastomosis of the homograft. PMID- 11414094 TI - Angioplasty for the treatment of buttock claudication caused by internal iliac artery stenoses. AB - A case of buttock claudication due to isolated internal iliac artery stenoses is presented. Although ankle systolic pressure at rest was within normal limits and distal pulses were palpable, an angiogram demonstrated severe stenoses. A computed tomography scan showed no spinal stenosis. The patient was successfully treated with angioplasty. This diagnosis may be elusive if ankle pressure or distal pulses are normal, thereby directing the clinician's suspicion away from vascular pathology. PMID- 11414095 TI - Inferior mesenteric artery aneurysm associated with occlusion of the superior mesenteric and celiac arteries. AB - We report a case of a large aneurysm of the inferior mesenteric artery that extended from its origin to bifurcation in the left colic and sigmoidal arteries, and was associated with occlusion of the celiac and superior mesenteric arteries in a 64-year-old patient. The diagnosis was made by arteriography. The patient underwent angioplasty and stenting of the superior mesenteric artery before the operation, which consisted of resection of the aneurysm and reimplantation of the left colic and the sigmoidal arteries separately in the aorta. We believe that this is the first reported case managed by angioplasty in combination with surgery, as well as reimplantation of the branches of the inferior mesenteric artery. PMID- 11414096 TI - Aortic thrombosis and ulcerative colitis. AB - Arterial thrombotic accidents in the course of inflammatory bowel disease are rare. They generally affect young adults whose disease is active. We observed a case of aortic and renal arterial thrombosis in a 40-year-old woman who was suffering from ulcerative colitis. Surgical thrombectomy ensured good postoperative results, without any ischemic or renal sequelae. Six days later the patient presented with distal thrombosis of the splenic artery, which receded under anticoagulant treatment. The physiopathology of thromboembolic events in the course of inflammatory bowel disease is uncertain. Such events result from a state of hypercoagulability of various mechanisms, which can be observed in active inflammatory bowel disease. This possibility of serious arterial thrombosis argues in favor of long-term anticoagulant treatment when inflammatory bowel disease is active. PMID- 11414097 TI - Chronic rupture of abdominal aortic aneurysm manifesting as crural neuropathy. AB - Chronic rupture of abdominal aortic aneurysm (AAA) resulting in unusual clinical manifestations can occur if the resistance of structures surrounding the aorta is sufficient to contain hemorrhage. In this report, we describe five cases of chronic ruptured AAA in which the presenting feature was crural neuropathy. All patients were male with a mean age of 74 +/- 1.8 years. At the time of presentation, crural neuropathy had been ongoing for 3 to 9 weeks. In three cases, AAA was not initially suspected because an inadequate clinical examination was performed (not in the vascular surgery department) and because of the small diameter of the aorta in relation to the patient's morphology. Two patients had one episode of hypotension that was wrongly attributed to vagal attack. Abdominal CT scanning was always diagnostic of chronic rupture. In two cases, rupture was associated with erosion of the body of one or more vertebrae and laboratory evidence of inflammation, i.e., increase in sedimentation rate and fibrinogen level. The mean diameter of the AAA was 7.1 +/- 0.9 cm (range 5-10 cm). All patients underwent midline laparotomy, which was performed under emergency conditions in two cases, under semi-emergency conditions in one case, and electively in two cases. Perforation was consistently located on the posterolateral wall of the aorta and varied from 1 to 3 cm in length. Repair was performed using an aortobifemoral prosthesis in four cases, and a straight tube in one case. The patient who underwent emergency surgery died 4 days after the procedure. The remaining four patients recovered uneventfully and were discharged after 10 days. In the elderly, ruptured AAA should be included in the differential diagnosis of crural neuropathy. An episode of hypotension, regardless of its duration, in an elderly patient should be given serious consideration as a possible sign of ruptured AAA with ongoing retroperitoneal hemorrhage. PMID- 11414098 TI - Surgical repair of an aneurysm of the innominate artery with fistulization into the trachea. AB - This report describes a 43-year-old patient presenting with an aneurysm located at the junction between the innominate artery and aorta, with spontaneous fistulization into the trachea. Emergency treatment of this unusually located lesion was undertaken to prevent intratracheal rupture. After ligation of the innominate artery, right common carotid artery, and right subclavian artery due to the risk of infection, the tracheal fistula was treated by direct closure and exclusion using an autologous pericardial flap. The procedure was performed under extracorporeal circulation and circulatory arrest with profound hypothermia. PMID- 11414099 TI - [Molecular approach in the diagnosis of human tuberculosis]. PMID- 11414100 TI - [Survey on giardiasis propagation in Pescara]. PMID- 11414101 TI - Appropriateness of admission and hospitalization days in a specialist hospital. AB - The assessment of hospital utilization is an important tool in the management of Hospital Health Care. The Authors examined the extent and the reasons of inappropriate hospital admission and stay in patients admitted to the Monaldi Hospital, Naples, Italy. Five hundred and thirty three medical records, were analysed. The survey was carried out in the period May-October 1999, using the italian version of the Appropriateness Evaluation Protocol (AEP) method. Of the admissions days, 15.8% were considered inappropriate; the most frequent cause of inappropriateness was waiting period for diagnostic test. The highest percentage of inappropriateness was found for females, for less serious pathologies and in the summer months. Compared to the admissions, the proportion of inappropriateness of the index days was higher (35.5%); the most frequent cause of an inappropriate day of stay was the persistence of mild symptoms which according to the attending physician justified prolonging the stay, while the same variables found for inappropriate admission (female sex, less serious pathologies and summer months) were related to inappropriateness of stay. This survey has allowed us to offer some useful suggestions with a view to making some of the activities of the "Monaldi Hospital" more efficient. PMID- 11414102 TI - [Teratogenic effects of environmental lead exposure: a population case-control study]. PMID- 11414103 TI - [Epidemiology study on the prevalence of psychiatric disorders among Sardinian immigrants in Paris compared with Parisians and Sardinians in Sardinia]. PMID- 11414104 TI - [Smoking among students from several schools in Lazio]. PMID- 11414105 TI - [Interregional Italy-Greece Project II: planning of the coastal monitoring by the Operative Unit No. 14 from Lecce]. PMID- 11414106 TI - [Assessment of the hygienic-sanitary conditions of the Lake Faro (ME) in the light of the D.L. vo 131/92]. PMID- 11414107 TI - [The risk of Escherichia coli O157:H7 in Sicily, through imported bovines for slaughter]. PMID- 11414108 TI - [Seroepidemiologic survey on varicella-zoster infection in the Province of Lecce]. PMID- 11414109 TI - [Nosocomial infections: from the hospital to integrated care. Structural, technologic, organizational, and economic features]. PMID- 11414110 TI - [Disinfection and sterilization: methodologies and indications]. PMID- 11414111 TI - [Nosocomial infections: epidemiologic models and prevention strategies]. PMID- 11414112 TI - [Infections and their control in dentistry]. PMID- 11414113 TI - [Role of infection control practitioners]. PMID- 11414114 TI - [Quality certification in the prevention and control of nosocomial infections]. PMID- 11414115 TI - [Extent of burn lesions--an analysis of 68 burned cadavers]. AB - The burned bodies (n = 68) autopsied at the Freiburg Institute of Forensic Medicine in the years 1996-2001 (until the end of February) were examined with regard to the extent of consumption by the fire using the classifications suggested by Eckert et al., Maxeiner, Glassman and Crow, and Gerling et al. Further classification parameters were exposure of the body cavities and amputation of extremities as a result of the fire. Most cases were accidental deaths (approximately 75%). The fires occurred mainly in buildings (57%) or vehicles (34%); in 6 cases the burning took place in the open air. In most cases the degree of consumption by the fire was slight to moderate. Only 15% of the burned bodies showed extensive destruction with severe loss of soft tissue and shrinking or amputation of the extremities. In one case the soft tissue was completely destroyed with fragmentation of the calcined skeletal remains. In bodies recovered from house fired the extent of consumption by the fire was usually less pronounced than in bodies retrieved from burned-out vehicles. PMID- 11414116 TI - [Was the German forensic psychiatrist Hans Szewczyk the first modern profiler?]. AB - The term "profiler" is usually associated with the FBI. For more than two decades a multitude of publications has appeared on this subject in the Anglo-American language area describing the work of psychiatrists and psychologists as profilers in spectacular crimes. This article deals with the forensic psychiatrist Prof. Dr. Dr. Hans Szewczyk, who probably worked out the first perpetrator profile on a scientific basis in 1969-71 already when three boys were murdered in Eberswalde (state of Brandenburg); his profile made an essential contribution to the solution of the case. PMID- 11414117 TI - [Early post-mortem animal bites after suicidal gunshot wound to the head of a police dog handler]. AB - A 41-year-old police officer committed suicide in his office by firing a contact shot to the left temple. When the body was found 12 hours later, the German shepherd dog the officer had been in charge of had caused considerable damage on the lower half of the face and the adjacent parts of the neck. The clothing of the suicide was undamaged. Blood traces in the shape of paws were found at the place of death. The wound margins of the postmortem animal injury were nicked and jagged by the canine teeth. The entrance and exit wounds were not affected by the soft tissue defect. The findings gathered are discussed in reference to the relevant literature. PMID- 11414118 TI - [Detection of cocaine in blood stains]. AB - Cocaine is rapidly degraded in blood samples, and its degradation was found to be highly dynamic in nature. The analysis of blood spots dried on filter paper may provide a method to minimize the break-down of cocaine and to largely preserve the analytical profile of the parent drug and its hydrolysis products at the time of sampling. The short term stability of cocaine in 100 microL blood spots prepared from unpreserved and preserved (sodium fluoride, 0.25%) blood samples was compared to the stability of the particular whole blood specimens stored in tubes at ambient temperature and at -20 degrees C. Due to dehydration, both the chemical and the enzymatic hydrolysis of cocaine and its products could be stopped in dried blood spots. More than 75% of the initial cocaine concentration could be detected in the blood spots, and the analytical profile was ensured for 17 days. Provided its practical suitability, the spot technology should offer a simple approach to detect actual impairment of motorists taken in police custody in the view of section 24a of the German traffic act as well as in cocaine associated criminal cases. PMID- 11414119 TI - [A case of fatal amniotic fluid embolism]. AB - A 31-year-old pregnant woman (multipara), who was near the expected date of birth, was found dead in the joint flat by her partner. She was lying beside the bed and showed burns on the front of the body caused postmortem by contact with a radiator. Autopsy revealed a rupture of the fetal membranes, the inner layers of the uterus and a large vein. After centrifugation, the supernatant of the blood from the right half of the heart was turbid and contained small corpuscular amniotic fluid components. The diagnosis of amniotic fluid embolism could be confirmed histologically. In the pulmonary vessels of the mother meconium, lanugo hairs and epidermic scales could be demonstrated; in addition the fetal horny lamellae were confirmed immunohistochemically (using monoclonal antibody against cytokeratin). PMID- 11414120 TI - [Suicide with a walking cane gun: weapon, injury picture and psychopathology]. AB - In Germany firearms camouflaged as items of daily use are forbidden. While reports on ball-point pen-guns have repeatedly been published in literature, the use of a walking-stick gun is a rarity. The report deals with the suicide of a 31 year-old schizophrenic man who inflicted a shotgun injury to his head with typical morphological characteristics. The weapon used was a normal-looking walking stick with a barrel, cal. .410. Under forensic and criminological aspects the significance of a walking-stick gun is comparable to that of ball-point pen guns. Both weapons can be carried and used inconspicuously, but from a walking stick gun also ammunition of larger calibres with a higher potential of injury can be fired. PMID- 11414121 TI - Studies of the molecular genetics of impaired insulin secretion and obesity. Two intermediary phenotypes predisposing to type 2 diabetes mellitus. PMID- 11414122 TI - Bordetella pertussis in the aetiology of chronic cough in adults. Diagnostic methods and clinic. AB - Cough is one of the most common complaints causing patients to seek medical attention, and chronic cough, defined as a cough period of at least three weeks, accounts for more than a third of referrals to a chest physician. Cough is an important factor in the spread and survival of microorganisms, but until recently little attention has been given to Bordetella pertussis (B. pertussis) in patients with chronic cough. This review summarizes the B. pertussis diagnostic methods--culture--polymerase chain reaction (PCR), and serology--and surveys the literature on B. pertussis and chronic cough in adults. There is growing evidence that B. pertussis is an important cause of persistent cough in adults; thus prevalence of pertusssis of 12.4-26% has been reported in studies from US, Australia and Germany. Recently we found evidence of pertussis as the cause of chronic cough in 16% of otherwise healthy adults in Denmark. Therefore, patients with chronic cough should be examined for B. pertussis infection. The demonstration of B. pertussis in an adult patient with chronic cough has two advantages: 1) the patient can be reassured that symptoms will disappear spontaneously, why more or less invasive examinations and empirical therapeutic trials can be omitted, 2) the source of infection can be eradicated, and contact persons, particularly non vaccinated infants in whom pertussis might be very severe, can be treated in order to avoid or attenuate clinical symptoms. PMID- 11414123 TI - Myocardial perfusion imaging and coronary angiography in patients with known or suspected stable angina pectoris. AB - INTRODUCTION: The patho-physiological cause of angina pectoris is myocardial ischaemia, which can be objectified by myocardial perfusion imaging (MPI). METHODOLOGY: MPI was undertaken prior to coronary angiography (CAG) in 86 randomly selected patients with known or suspected stable angina pectoris. RESULTS: Among 78 adequately stressed patients, MPI was normal in 28 (36%) and showed reversible and irreversible perfusion abnormalities in 30 (38%) and 20 patients (26%), respectively. Coronary angiograms were normal in 28 (36%) and revealed at least one > or = 50% stenosis in 50 patients (64%) (16 with single and 34 with multi vessel disease). Using angiography as a reference, the sensitivity and specificity of MPI in detecting coronary artery disease was 88% and 93%, respectively. DISCUSSION: MPI demonstrates regional hypoperfusion whereas CAG depicts anatomical stenosis in epicardial arteries. Both modalities are potentially relevant in patients with stable angina pectoris. The functional significance of coronary artery lesions is, however, variable and MPI can demonstrate normal myocardial perfusion in the presence of moderate lesions. MPI exhibited a high sensitivity and specificity regarding significant lesions. More than one third of the subjects had a normal MPI and a normal CAG. Patients with stable angina pectoris and a normal MPI have a very low risk of cardiac events and do usually not require further invasive investigation or therapy. Reversible ischaemia and irreversible ischaemia with demonstration of viable tissue call for coronary revascularisation. PMID- 11414124 TI - Penicillin as empirical therapy for patients hospitalised with community acquired pneumonia at a Danish hospital. AB - INTRODUCTION: We report on the outcome of a study of patients hospitalised with community acquired pneumonia (HCAP) at a Danish university hospital. METHODOLOGY: In a retrospective study of 243 consecutive patients with radiographically verified HCAP, data on clinical and laboratory findings and outcome parameters were collected. Three groups were established according to the initial choice of antibiotic(s): penicillin only (n = 160); non-allergic patients starting broader spectrum therapy (n = 54); and patients with suspected penicillin allergy (n = 29). RESULTS: The overall mortality within three months was 12% and the readmission rate within three months was 20%. The three treatment groups were comparable with respect to most demographic and clinical criteria at baseline. No significant differences in outcome between the groups were found: the mortality was 12.5%, 13.0%, and 10.3%, respectively, p = 0.94, and the readmission rate 20.3%, 24.0%, and 14.8%, respectively; p = 0.63. CONCLUSION: Patients treated for community-acquired pneumonia at a Danish university hospital had clinical outcomes fully at height with findings from other countries, and half of the patients were successfully treated with penicillin monotherapy. No differences in clinical outcomes were documented between patients treated empirically with broad spectrum therapy and penicillin monotherapy. Therefore, penicillin seems to be a reasonable first choice for initial therapy of HCAP in Denmark as in other regions with similar patterns of microbial pathogens and resistance. PMID- 11414125 TI - Paracetamol poisoning among immigrants in a department of hepatology. AB - INTRODUCTION: An increased incidence of suicides and suicidal behaviour among immigrants has been described in other countries. In Denmark, misuse of paracetamol is suspected in some foreign-born minority groups, although no data have been produced to substantiate this suspicion. METHODOLOGY: A retrospective study of the incidence of paracetamol poisoning in patients admitted to a specialised department of hepatology from 1994 to 1999 was carried out. RESULTS: Of a total of 580 patients, 56 (9.7%; 95%-confidence interval 7.2-12.1%) were immigrants, among whom a significant overrepresentation was found of immigrants from Turkey, Iran, Pakistan, and Lebanon (Observed/Expected-ratios of 1.95, 4.14, 2.67, and 2.45 respectively; p < 0.05). The immigrants differed from the Danish born patients being younger (21 vs 35 years of age; p < 0.05), having a lower level of alcohol consumption (3% vs 30% with regular alcohol abuse; p < 0.05), and in general being less severely intoxicated (3% vs 22% developing hepatic encephalopathy; p < 0.05). Compared to the Danish-born patients, the immigrants more frequently stated socio-economic problems as the reason for their self poisoning (29% vs 10%; p < 0.05). CONCLUSIONS: The study demonstrates an overrepresentation of immigrants among patients admitted with paracetamol poisoning in Denmark. PMID- 11414126 TI - The Danish Multiple Sclerosis Registry. History, data collection and validity. AB - The Danish Multiple Sclerosis Registry was formally established in 1956 but started operating in 1949 with a nationwide prevalence survey. Since then, the Registry has continued collecting data on new and old cases of multiple sclerosis (MS) or suspected MS from multiple sources. The Registry reclassifies cases according to standardized diagnostic criteria (currently those of Poser et al). A total of 14,441 cases fulfilling the diagnostic criteria had been registered at the most recently completed follow-up by 1 January 1997; 10,851 had onset from 1948 to 1996 and 3560 before 1948. The completeness has formerly been estimated at about 90%, higher for cohorts with older onset and lower for cohorts with onset close to follow-up. The estimated validity of the diagnosis for autopsy cases classified as definite MS in the Registry is 94%. A long-term nationwide Registry has proved to be a valuable instrument for monitoring incidence and prevalence, analysing survival, performing genetic analysis, providing unselected patient samples for clinical analyses, performing case-control studies and prospective studies and estimating the need for treatment and care. PMID- 11414127 TI - Asymmetries in visual search: an introduction. AB - In visual search tasks, observers look for a target stimulus among distractor stimuli. A visual search asymmetry is said to occur when a search for stimulus A among stimulus B produces different results from a search for B among A. Anne Treisman made search asymmetries into an important tool in the study of visual attention. She argued that it was easier to find a target that was defined by the presence of a preattentive basic feature than to find a target defined by the absence of that feature. Four of the eight papers in this symposium in Perception & Psychophysics deal with the use of search asymmetries to identify stimulus attributes that behave as basic features in this context. Another two papers deal with the long-standing question of whether a novelty can be considered to be a basic feature. Asymmetries can also arise when one type of stimulus is easier to identify or classify than another. Levin and Angelone's paper on visual search for faces of different races is an examination of an asymmetry of this variety. Finally, Previc and Naegele investigate an asymmetry based on the spatial location of the target. Taken as a whole, these papers illustrate the continuing value of the search asymmetry paradigm. PMID- 11414128 TI - Curvature discontinuities are cues for rapid shape analysis. AB - Using the visual search method, we show that stimuli that contain curvature discontinuities (i.e., points where the second derivative along an image contour is not defined) are easily found among stimuli containing only smooth changes in curvature. Curved stimuli that lack curvature discontinuities, however, are difficult to find among distractors that have them. These results suggest that the visual system detects and analyzes abrupt changes in curvature in the image quickly to extract vital information about the 3-D structure of the visual environment. PMID- 11414129 TI - Detection of shape orientation depends on salient axes of symmetry and elongation: evidence from visual search. AB - Three experiments investigated the role of the global spatial structure of two dimensional (2-D) shapes in terms of symmetry and elongation on visual search for shape orientation. Experiment 1 demonstrated the often reported orientation search asymmetry (i.e., a faster detection of a tilted target among vertical distractors than the reverse) for the global orientation of 2-D polygons that possess a salient, "principal" axis of symmetry or elongation. Moreover, the search asymmetry depended on the orientation of the principal axis, rather than on the orientation of local contours. Further exploration of this effect with polygons (Experiment 2) showed that the search asymmetry for global orientation occurred for shapes containing an axis of symmetry; elongation, on the other hand, did not seem to be crucial. Finally, Experiment 3 demonstrated orientation search asymmetries with shapes composed of curved rather than straight contours: Here, the search asymmetry occurred as a function of the orientation of both axes of symmetry and elongation. Overall, search for global orientation was less efficient than search for local orientation. The results suggest that the perception of the global orientation of shapes is mediated by axis-based descriptions in terms of perceptually salient axes of symmetry and elongation. PMID- 11414130 TI - Visual search for a socially defined feature: what causes the search asymmetry favoring cross-race faces? AB - Levin (1996, 2000) reported that white subjects search for black targets more quickly than they search for white targets, suggesting that black faces are perceived as having a feature that is lacking in white faces. Here we test one of the implications of this asymmetry by having subjects search for same-race (SR) and cross-race (CR) faces that are distorted to look less like each other (producing caricatures that enhance race-specifying features), or are distorted to look more like each other (a prototypical distortion expected to reduce the salience of race-specifying features). Experiments 1 and 2 show that caricaturing the feature-positive CR distractors speeds search for the SR face and that prototypical distortion slows this search. The same distortions in SR faces did not affect the search slopes. However, these distortions also eliminated the overall advantage for CR faces. Experiment 3 shows that trial-to-trial variation in the specific distractors in each display can eliminate the asymmetry and suggests that this asymmetry depends on the subjects' ability to set a consistent a priori perceptual criterion when searching for a CR target, while the distortion effects emphasize the importance of distractor-rejection processes in determining the form of a serial search asymmetry. PMID- 11414131 TI - Visual search asymmetries in motion and optic flow fields. AB - In visual search, items defined by a unique feature are found easily and efficiently. Search for a moving target among stationary distractors is one such efficient search. Search for a stationary target among moving distractors is markedly more difficult. In the experiments reported here, we confirm this finding and further show that searches for a stationary target within a structured flow field are more efficient than searches for stationary targets among distractors moving in random directions. The structured motion fields tested included uniform direction of motion, a radial flow field simulating observer forward motion, and a deformation flow field inconsistent with observer motion. The results using optic flow stimuli were not significantly different from the results obtained with other structured fields of distractors. The results suggest that the local properties of the flow fields rather than global optic flow properties are important for determining the efficiency of search for a stationary target. PMID- 11414132 TI - Target-tilt and vertical-hemifield asymmetries in free-scan search for 3-D targets. AB - In this study, asymmetries in finding pictorial 3-D targets defined by their tilt and rotation in space were investigated by means of a free-scan search task. In Experiment 1, feature search for cube tilt and rotation, as assessed by a spatial forced-choice task, was slow but still exhibited a characteristic "flat" slope; it was also much faster to upward-tilted cubes and to targets located in the upper half of the search field. Faster search times for cubes and rectangular solids in the upper field, an advantage for upward-tilted cubes, and a strong interaction between target tilt and direction of lighting (upward or downward) for the rectangular solids were all demonstrated in Experiment 2. Finally, an advantage in searching for tilted cubes located in the upper half of the display was shown in Experiment 3, which used a present-absent search task. The results of this study confirm that the upper-field bias in visual search is due mainly to a biased search mechanism and not to the features of the target stimulus or to specific ecological factors. PMID- 11414133 TI - The effect of familiarity on visual-search performance: evidence for learned basic features. AB - In this study, the effects of familiarity on visual search were investigated. To avoid any confounding between familiarity and visual-feature differences between item pairs, Ns and mirror-Ns were presented as target and as distractors to a group of German participants and to a group of Slavic participants. For the Germans only N was familiar, whereas for the Slavs both N and mirror-N were familiar. The results show that search was difficult only when the Germans had to find an N among mirror-Ns. In any other case, search was efficient. Therefore, our results demonstrate that, contrary to earlier suggestions, search for a familiar item among familiar distractors can be easy. This supports the hypothesis that familiarity improves distractor grouping. However, the data are also compatible with the idea that letters are standard or basic features, which implies that basic features can be learned. PMID- 11414134 TI - Visual search asymmetry: the influence of stimulus familiarity and low-level features. AB - Wang, Cavanagh, and Green (1994) demonstrated a pop-out effect in searching for an unfamiliar target among familiar distractors (U-F search) and argued for the importance of a familiarity difference between the target and the distractors in determining search efficiency. In four experiments, we explored the generality of that finding. Experiment 1 compared search efficiency across a variety of target distractor pairs. In Experiments 2, 3, and 4, we used Chinese characters and their transforms as targets and distractors and compared search performance between Chinese and non-Chinese participants. We demonstrated that search asymmetry and search efficiency in the U-F condition are influenced by the presence of low-level feature differences between the familiar and the unfamiliar stimuli. Our findings suggest that the familiarity of the distractors, rather than the familiarity difference between the target and the distractors, determines search efficiency. We also documented a counterintuitive familiarity inferiority effect, suggesting that knowledge of search stimuli may, sometimes, be detrimental to search performance. PMID- 11414135 TI - Search asymmetries? What search asymmetries? AB - In order to establish a search asymmetry, one must run an experiment with a symmetric design and get asymmetric results. Given an asymmetric design, one expects asymmetric results, and such results do not imply an asymmetry in the search mechanisms. In this paper, I argue that a number of experiments purporting to show search asymmetries contain built-in design asymmetries. A saliency model of visual search predicts the results of these experiments, using only a simple measure of target-distractor similarity, without reliance on asymmetric search mechanisms. These results have implications for search mechanisms and for other experiments purporting to show search asymmetries. PMID- 11414136 TI - The influence of color on transient system activity: implications for dyslexia research. AB - Metacontrast and apparent motion experiments designed to utilize transient system resources were adopted to investigate the proposal that transient system activity is differentially influenced by different colored stimuli. The results generally showed no effect of color on transient system activity in either adults or children. However, the predicted pattern of results was demonstrated when contrast rather than color was manipulated in a final metacontrast experiment. We discuss the tenuousness of the proposal that color differentially influences transient activity, exploring its physiological implications and its durability as a theory of transient activity regarding reading-disability research. PMID- 11414137 TI - Strength of German accent under altered auditory feedback. AB - Borden's (1979, 1980) hypothesis that speakers with vulnerable speech systems rely more heavily on feedback monitoring than do speakers with less vulnerable systems was investigated. The second language (L2) of a speaker is vulnerable, in comparison with the native language, so alteration to feedback should have a detrimental effect on it, according to this hypothesis. Here, we specifically examined whether altered auditory feedback has an effect on accent strength when speakers speak L2. There were three stages in the experiment. First, 6 German speakers who were fluent in English (their L2) were recorded under six conditions -normal listening, amplified voice level, voice shifted in frequency, delayed auditory feedback, and slowed and accelerated speech rate conditions. Second, judges were trained to rate accent strength. Training was assessed by whether it was successful in separating German speakers speaking English from native English speakers, also speaking English. In the final stage, the judges ranked recordings of each speaker from the first stage as to increasing strength of German accent. The results show that accents were more pronounced under frequency-shifted and delayed auditory feedback conditions than under normal or amplified feedback conditions. Control tests were done to ensure that listeners were judging accent, rather than fluency changes caused by altered auditory feedback. The findings are discussed in terms of Borden's hypothesis and other accounts about why altered auditory feedback disrupts speech control. PMID- 11414138 TI - Accenting and detection of timing variations in tone sequences: different kinds of accents have different effects. AB - The effect of intensity and pitch accents on the perception of timing was examined in two experiments using a signal detection procedure. Analyses of sensitivity and response bias revealed opposite effects of intensity and pitch accents under similar conditions. Time intervals preceding intensity accents were perceived as longer, but time intervals preceding pitch accents were perceived as shorter. These results showed that listeners found it easier to detect timing variations that were contrary to expectations, as compared with variations that were consistent with expectations. In the present case, listeners should have expected shorter time intervals before intensity accents and longer intervals before pitch accents. The fact that the effects were observed with stimuli that had minimal musical structure demonstrated the contribution of psychoacoustic factors to such phenomena. PMID- 11414139 TI - Temporal bisection with trial referents. AB - In temporal bisection, the subject is required to compare the similarity of a probe duration with two referent durations. Two bisection experiments with human subjects are reported in which performance under three referent conditions was compared. In two of the conditions, no-referent and fixed-referent, the referent pair was constant throughout the session and was varied between sessions. In the no-referent condition, samples of the referents were presented at the beginning of a block of trials, but not on the probe trials. In contrast, in the fixed referent condition, the referent pair was presented on each probe trial. In the third referent condition, roving-referent, the referent pair was also presented on each probe trial, but the values of the pair varied during a session rather than between sessions. The pseudologistic Model (PLM; Killeen, Fetterman, & Bizo, 1997) provided an excellent account of the data from all referent conditions and indicated that (1) the scalar variability was located in the perception of the probe and was independent of the structure of the bisection task, (2) the role of the referents was to set the criterion, (3) the probe was compared with the criterion rather than with the referents, and (4) the comparison of the probe with the criterion occurred even when trial referents were available. PMID- 11414140 TI - Reference frames and haptic perception of orientation: body and head tilt effects on the oblique effect. AB - The aim of this study was to examine the effect of body and head tilts on the haptic oblique effect. This effect reflects the more accurate processing of vertical and horizontal orientations, relative to oblique orientations. Body or head tilts lead to a mismatch between egocentric and gravitational axes and indicate whether the haptic oblique effect is defined in an egocentric or a gravitational reference frame. The ability to reproduce principal (vertical and horizontal) and oblique orientations was studied in upright and tilted postures. Moreover, by controlling the deviation of the haptic subjective vertical provoked by postural tilt, the possible role of a subjective gravitational reference frame was tested. Results showed that the haptic reproduction of orientations was strongly affected by both the position of the body (Experiment 1) and the position of the head (Experiment 2). In particular, the classical haptic oblique effect observed in the upright posture disappeared in tilted conditions, mainly because of a decrease in the accuracy of the vertical and horizontal settings. The subjective vertical appeared to be the orientation reproduced the most accurately. These results suggest that the haptic oblique effect is not purely gravitationally or egocentrically defined but, rather, depends on a subjective gravitational reference frame that is tilted in a direction opposite to that of the head in tilted postures (Experiment 3). PMID- 11414141 TI - Locus of the redundant-signals effect in bimodal divided attention: a neurophysiological analysis. AB - We reanalyzed the data from the study of Lamarre, Busby, and Spidalieri (1983). In that study, the activity of single neurons in area 4 of the motor cortex was recorded during a bimodal detection task in which a monkey (Macaca mulatta) had to respond as quickly as possible to a visual or an auditory signal or to both (redundant trials). Manual responses on redundant trials were speeded by the presence of both signals, as is typically found. The times between signal onsets and the first changes in neuronal activity were also speeded by redundant signals, but there was no difference between redundant-signals and single-signal trials in the time between the change in neuronal activity and movement onset. These results suggest that late motor processes are not speeded by redundant signals in bimodal detection tasks. PMID- 11414142 TI - Individual differences and the use of nonspecifying variables in learning to perceive distance and size: comments on McConnell, Muchisky, and Bingham (1998). AB - McConnell, Muchisky, and Bingham (1998) showed that observers are able to judge the distance and size of falling, rolling, and swinging balls and that performance improves after practice with feedback. They concluded that observers use information that specifies the spatial scales of the different event types- namely, event duration in combination with event-specific constants. The improvement was interpreted as the calibration of the event-specific constants. We argue that their analyses should have considered the use of optical variables that do not specify the to-be-perceived metrics and individual differences in variable use. Furthermore, we propose convergence on the more useful variables as an alternative explanation for the observed improvement. The viability of these arguments is demonstrated with an experiment in which participants are trained with feedback to judge the distance and size of freely falling balls. PMID- 11414143 TI - Commentary on Jacobs and Michaels (2001): calibration and perceptual learning in event perception. AB - Jacobs and Michaels (2001) have argued that increased precision in judgments of the viewing distance to a perceived event should be attributed in part to perceptual learning. They found that observers used feedback to attune to the appropriate information variables gradually. McConnell, Muchisky, and Bingham (1998) had found that observers used feedback to calibrate event-specific scaling coefficients, that the calibration of one type of event generalized to other types, and that calibration occurred suddenly. We argue that Jacobs and Michaels must be partially correct and that, in our experiments, both calibration and perceptual attunement were required for accurate and precise judgments. PMID- 11414144 TI - [Interdisciplinary vascular center: a method for consequential process optimization in the hospital]. PMID- 11414145 TI - [Molecular radiology. II: Molecular imaging]. AB - The most important strategies for molecular imaging are presented. Processes can now be imaged in vitro and in vivo at the molecular level with the help of modern procedures such as SPECT, PET, MRI, and highly developed optical methods. The ability to view the expression of genes, the pharmacokinetics of gene therapy vectors, of therapeutic DNA, and of classical pharmacons in vivo opens up completely new perspectives for the research on, diagnosis of, and therapy for diseases. The current status of these developments are described, potential fields of use and possibilities for further development are outlined. PMID- 11414146 TI - Cardiac metabolism and function in patients with multiple sclerosis: a combined 31P-MR-spectroscopy and MRI study. AB - OBJECTIVE: Involvement of the autonomic system in multiple sclerosis (MS) may concur with dysfunction of the cardiovascular system. The introduction of potentially cardiotoxic immunosuppressive drugs like Mitoxantrone into the treatment of MS warrants proper assessment of preexisting heart disease. However, systematic analyses of functional and metabolic derangements in MS are missing. Using quantitative 31P-MR-spectroscopy (MRS) and MR-imaging (MRI) metabolic and functional parameters were analyzed in patients with MS in comparison to healthy volunteers. SUBJECTS/METHODS: 14/15 patients with MS could be included in the study, as the MRS examination of one patient had to be excluded from analysis due to movement during the examination. Using chemical shift imaging (CSI) and AMARES, phosphocreatine (PCr) to adenosine triphosphate (ATP) ratios, characterizing myocardial high-energy phosphate metabolism, were determined. Additionally, absolute concentrations of PCr and ATP were calculated by SLOOP (Spatial Localization with Optimal Pointspread Function). Analysis of functional changes was performed by Cine-MRI. 14 healthy volunteers matched for age and gender served as control. RESULTS: A significant decrease of absolute PCr concentration was observed in patients with MS compared to matched volunteers (p < 0.05), whereas ATP concentrations showed no significant changes (p = 0.27). Metabolite ratios calculated by SLOOP or AMARES showed a tendency to be reduced in patients, however, did not reach significance (p = 0.08, SLOOP; p = 0.47, AMARES). Using volunteers' mean values +/- 2 x SD as cut off value revealed PCr changes in 5 of 14 patients, whereas only 2 also had pathologic PCr/ATP ratios. Functional analysis by MRI depicted depressed left ventricular ejection fraction in 4 patients. CONCLUSIONS: The reduction in cardiac high-energy phosphates in some patients with MS points to a subclinical involvement of the heart. This may be important for treatment with potentially cardiotoxic drugs. Longitudinal studies are need to understand the clinical relevance of our findings. PMID- 11414147 TI - [MR angiography of pelvic and leg vessels with automatic table movement technique ("Mobi-Trak")--clinical experience with 450 studies]. AB - PURPOSE: By a retrospective clinical evaluation of 450 MR angiograms of the lower extremity arteries the question was posed of whether MR angiography can replace i.a. DSA. MATERIAL AND METHODS: Image quality of 450 MR angiograms was graded by 4 radiologists; in 102 patients who underwent radiological intervention a comparison between MR angiogram and i.a. DSA was done. Additionally, MR angiography was compared with the results of surgical intervention in 106 cases. RESULTS: 6% of 450 examinations showed a significant limitation of diagnostic information. 5 examinations (1%) had to be redone by i.a. DSA. In all cases the diagnosis from MRA was proven by the results of the following i.a. DSA or surgical procedure. CONCLUSION: The retrospective clinical evaluation of 450 examinations demonstrated that MR angiography of the lower extremity arteries can replace i.a. DSA in routine use. PMID- 11414148 TI - [Dynamic MR defecography for diagnosis of combined functional disorders of the pelvic floor in proctology]. AB - PURPOSE: Evaluation of magnetic resonance defecography in the diagnosis of pelvic floor disorders were examined prospectively. MRI was performed on a 1.5 T scanner. The rectum was opacified with 200 ml of ultrasound transmission gel. A sagittal single section T2-weighted gradient echo sequence with a temporal resolution of 1.1 second was performed. Changes of the anorectal angle and the position of the pelvic organs in relation to the pubococcygeal line were registered at rest, during straining, and during evacuation of the rectum. RESULTS: Patients with obstructed defecation (n = 15) showed prolapse of the rectal mucosa (n = 5), anterior rectocele (n = 8), pelvic floor descent (n = 5), enterocele (n = 2), and anorectal dyscoordination (n = 3). Individuals with stool incontinence (n = 15) had an anterior rectocele (n = 10), pelvic floor descent (n = 11), enterocele (n = 2), prolapse of rectal mucosa (n = 1), and a puborectal insufficiency (n = 1). Urine incontinence was associated with cystocele (n = 10) or normal findings (n = 4). In patients with unspecific symptoms (n = 6), anorectal dyscoordination (n = 4) and prolapse of the rectal mucosa (n = 2) were found. MRI was superior for the detection of enteroceles, cystoceles and pelvic floor descent compared with clinical investigation. CONCLUSION: Dynamic MR imaging supplies complex information in anorectal disease and thus improves proctoscopy. PMID- 11414149 TI - [Value of selective MIP reconstructions in respiratory triggered 3D TSE MR cholangiography on a workstation in comparison with MIP standard projections and single-shot MRCP]. AB - PURPOSE: Comparison of anatomical visualisation and diagnostic value of selective MIP reconstructions of respiratory triggered 3D-TSE-MRCP versus standard MIP reconstructions and single-shot MRCP. MATERIAL AND METHODS: 50 patients with pancreaticobiliary disease were examined at 1.5 Tesla (ACS NT II, Philips Medical Systems) using a breath-hold single-shot (SS) and a respiratory triggered 3D-TSE MRCP technique in 12 standard MIP projections. Additional selective MIP reconstructions with different slice thickness (2, 4, 10 cm) and projections were performed on a workstation. Visualization of the pancreaticobiliary system and the diagnostic value of the examinations were analysed. RESULTS: Single-shot and 3D-TSE in standard projections showed comparable anatomical visualisation. On selective MIP reconstructions the biliary system (SS p < 0.002; 3D-TSE p < 0.000) and the periampullary region (SS p < 0.000; 3D-TSE p < 0.003) were more clearly seen than on SS and standard MIP reconstructions. Furthermore, superior visualisation of the pancreatic duct could be achieved with additional selective MIP reconstructions in contrast to standard MIP (p < 0.003). Sensitivity and diagnostic accuracy showed superior results for selective and standard MIP reconstructions, but no significant differences between the three techniques were found. CONCLUSION: SS and standard MIP reconstructions showed comparable anatomical visualisation. Selective MIP postprocessing on a workstation offers a better visualisation of the pancreaticobiliary system and is useful for detecting pathological alterations. PMID- 11414150 TI - [Incidence and significance of small focal liver lesions in MRI]. AB - PURPOSE: Analysis of the frequency and significance of small focal liver lesions (< or = 2 cm) detected on MRI in the presence or absence of a history of malignancy. METHODS: 628 MRI examinations of the liver performed during 1994-1996 were evaluated. The inclusion criterion into the study was the detection of a focal liver lesion with a size < or = 2 cm. The frequency, the size, the diagnostic proof, and the differential diagnosis of the focal liver lesions were analysed with regard to the patients history of a known malignant tumor. RESULTS: Overall, 179 of the 628 patients (28.5%) had focal liver lesions < or = 2 cm (n = 338). 58.9% of the lesions could be classified based upon follow-up studies by ultrasound, CT or MRI, or by biopsy. The remaining 41.1% of the lesions could not be classified due to the absence of follow-up examinations. 57.3% of all proven lesions were benign and 42.7% were malignant. A history of a malignant tumor was present in 76.7% of all patients with small liver lesions; however, lesions were benign in these patients in 50.6% of the cases. In patients with no known history of a malignancy, 75% of the lesions were benign and 25% were malignant. However, these malignant lesions were in 10/11 cases hepatocellular carcinomas in patients with liver cirrhosis. CONCLUSION: Even in the presence of a history of a malignant tumor, about 50% of the detected small liver lesions on MRI are benign. In the absence of a tumor history the probability of a small malignant liver lesion is very low, if the patients does not have liver cirrhosis where small hepatocellular carcinomas can be present. PMID- 11414151 TI - [Value of magnetic resonance tomography for interventions in the ENT specialty]. AB - PURPOSE: Presentation of new concepts and applications of MR-guided head and neck surgery are presented. Examples of diagnostic and therapeutic procedures such as evaluation of transseptal tumor biopsies, placement of afterloading catheters for brachytherapy, and microscopic surgery of paranasal sinuses in the open MRI are discussed. MATERIAL AND METHODS: 24 MRI-guided ENT-procedures (14 transsphenoidal biopsies, one transnasal biopsy, 6 placements of brachytherapy catheters, and 3 operations of the paranasal sinuses) were performed in an open 0.5 T MR system. RESULTS: Localisation and/or extension of all lesions as well as the placement of biopsy needles or catheters were determined with great precision during the interventions. CONCLUSIONS: Surgical risk and postoperative morbidity are significantly reduced in MR-guided surgery of the petroclival region and the region of head and neck compared to other, conventional methods. Thus, interventional MRI-guidance optimizes minimal invasive surgery and catheter placement in difficult anatomical regions like the petroclival region. PMID- 11414152 TI - [Obstruction of surgical portosystemic shunts in the early postoperative phase: interventional treatment by angioplasty with stent implantation]. AB - PURPOSE: Recurrent variceal bleeding in patients treated with surgical porto systemic shunting is most often due to shunt stenoses or occlusion. Radiological interventional procedures are a possible method of therapy and our experience herein is described in this report. PATIENTS AND METHODS: From 1997 to 1999 54 patients with recurrent variceal bleeding were treated with a surgical porto systemic shunt procedure. Of these early shunt occlusion occurred in 5 patients, which was treated with percutaneous transcatheter techniques. RESULTS: In only one patient was PTA alone sufficient to reestablish shunt patency, in four patient stent placement was necessary in addition. In the follow-up period 1 patient died 26 month after intervention with (autopsy-proven) patent shunt, in one patient shunt reocclusion occurred after 11 months and in 3 patients the shunt is still patent. CONCLUSIONS: PTA, if necessary in combination with stent placement, is an attractive alternative method of therapy in case of an early surgical porto-systemic shunt occlusion. PMID- 11414153 TI - [Percutaneous transfemoral implantation of a new, flexible thoracic aorta endoprosthesis using a percutaneous suture system for vascular suture--initial experiences]. AB - PURPOSE: To evaluate the transfemoral placement of a new, flexible stent-graft into the thoracic aorta and the suture-mediated closure of the femoral access. PATIENTS AND METHODS: Five patients were treated endovascularly with a stent graft for an aneurysm (n = 3) or acute dissection (n = 2) of the thoracic aorta via a femoral 24 F sheath. The femoral access site was closed with two suture mediated closure devices after placement of the stent-graft. RESULTS: The aneurysm or the false lumen was excluded from perfusion by the placement of the stent-graft in all patients. Hemostasis at the femoral access site was successful in all patients with the percutaneous suture device. A minor stenosis of the femoral artery was found angiographically in four patients after suture-mediated closure. Besides a reversible renal failure due to the medically induced hypotension for the treatment of an acutely ruptured aneurysm, no complications resulted from the stent-graft placement or the percutaneous suture. CONCLUSION: The percutaneous transfemoral placement of stent-grafts in the thoracic aorta using a suture-mediated closure of the access site is technically feasible. Long term results of the technique have to be awaited. PMID- 11414154 TI - [Endovascular therapy of renal artery stenosis: technical results with the Palmaz Corinthian stent]. AB - OBJECTIVE: To evaluate the technical performance and delivery characteristics of the Palmaz-Corinthian stent for endovascular therapy of atherosclerotic renovascular disease. METHODS: 61 patients underwent implantation of 76 Palmaz Corinthian (PC) stents in 72 arteries. 50 original PC and 26 PC stents with the modified IQ-design were employed. The indications comprised primary stenting of ostial (n = 49) or truncal (n = 1) stenosis or occlusion (n = 3), and selective stenting following complicated (dissection, n = 4) or unsuccessful (n = 8) angioplasty. The remaining stents were placed in patients with recurrent stenosis (n = 5) or acute aortic dissection (n = 2) involving the renal artery. Mean severity and length of stenosis were 81.3% and 9.8 mm, respectively. 39 lesions were rated eccentric or calcified. Data on technical success, complication rate, delivery characteristics and ease of placement compared to standard renal stents were retrieved from a prospective multicenter registry. RESULTS: Stent delivery was successful in all patients, major complications were not reported. Stent placement was suboptimal in 7 of 72 cases: 4 stents were located too distally in the renal artery, necessitating proximal coaxial overstenting in 2 cases. The distal part of the stenosis was incompletely covered and the orifice of a segmental branch inappropriately overstented in one case each. One stent was dislodged from the balloon, resulting in stent protrusion in the aortic lumen. Significant residual stenosis after stenting was not observed. Overall stent deliverability, trackability and potential repositioning inside the stenosis were rated positive, radio-opacity was rated superior for the IQ design. CONCLUSION: Technical performance and delivery characteristics of the Palmaz-Corinthian stent have been significantly improved compared to the Palmaz design, allowing mostly correct placement in renal artery stenoses with a low complication rate. PMID- 11414155 TI - [MRI in chronic epicondylitis humeri radialis using 1.0 T equipment--contrast medium administration necessary?]. AB - PURPOSE: Evaluation of the diagnostic value and confidence of contrast-enhanced MR imaging in patients with lateral epicondylitis in comparison to clinical diagnosis. MATERIAL AND METHODS: 42 consecutive patients with clinically proven chronic lateral epicondylitis and 10 elbow joints of healthy controls have been examined on a 1.0 T MR-unit. Criteria for inclusion in the prospective study were: persistent pain and a failed conservative therapy. The MR protocol included STIR sequence, a native, T2-weighted, fat-suppressed TSE sequence, and a Flash-2 D sequence. Also, fat-suppressed, T1-weighted SE sequences before and after administration of Gd-DTPA contrast media have been recorded. RESULTS: In 39/42 patients the STIR sequence showed an increased SI of the common extensor tendon. Increased MR signal of the lateral collateral ligament combined with a thickening and a partial rupture or a full thickness tear have been observed in 15/42 cases. A bone marrow edema at the lateral epicondilus was noticed in 6 of the studied patients and a joint effusion in 18/42 patients. After administration of contrast media we noticed an average increase of SI by about 150%. However, enhanced MR imaging did not provide additional information. CONCLUSION: In MR imaging of chronic epicondylitis administration of gadolinium-DTPA does not provide additional information. PMID- 11414156 TI - [Artefact reducing in diagnosis of lung embolism using spiral CT with saline bolus]. AB - PURPOSE: To improve the diagnostic efficacy of bolus-enhanced spiral CT (SCT) in the detection of pulmonary embolism using a saline push immediately after bolus injection of the contrast medium. PATIENTS AND METHODS: The study included 90 patients with suspected acute or chronic pulmonary embolism. The CT scan was performed in a caudocephaled direction. In Group I (n = 60) we applied a bolus contrast injection (120 ml, 3 ml/s, 300 mg J/ml), after a median delay of 25 s. Group II (n = 30) had the same contrast injection which was immediately followed by an additional saline push (60 ml, 2 ml/s). Streak artifacts originating from high contrast concentrations in the superior vena cava were rated on a 4-point scale for different locations: right pulmonary artery, pars basalis, truncus anterior, and the segmental upper lobe arteries. RESULTS: The incidence of artifacts in group I was nearly twice as high as in group II. The difference was significant (p < 0.05) for the upper and anterior superior lobe artery, the right pulmonary artery and the pars basalis. CONCLUSION: The presented protocol significantly reduces artifacts mainly by a washout of contrast medium in the superior vena cava. PMID- 11414157 TI - [Dose reduction in spiral-CT: detection of pulmonary coin lesions with and without anatomically adjusted modulation of tube current]. AB - PURPOSE: To investigate the potential of projection angle-dependent anatomical adaptation of tube current to reduce dose in spiral CT examinations of the thorax without loss in detectability of pulmonary nodules and image quality. PATIENTS AND METHODS: Dose can be reduced for non-circular patient cross-sections without an increase in noise if the tube current is reduced at those angular positions where the patient diameter and, consequently, attenuation are small. The examinations were dose with SOMATOM Plus 4 (Siemens AG). CT projection data were analysed to determine the optimum tube current for each projection angle in real time. We compared image quality, done, and detection rates of pulmonary nodules for thoracic spiral CT examinations with and without online anatomically adapted tube current control in a group of 38 patients. Three radiologists counted all intrapulmonary nodules in consensus separated in three different groups (< 5 mm, 5-10 mm, > 10 mm). Image quality was evaluated in a scale from 1-3 (1 = very good, 2 = good, 3 = worse). RESULTS: On average, the dose was reduced by 21% (15 34%). With a constant tube current, 704 pulmonary nodules were detected, with tube current modulation 707 pulmonary nodules. For three patients we saw more pulmonary nodules (five) with tube current modulation, for two patients we saw less pulmonary nodules (two). In a direct comparison, the missed intrapulmonary nodules were also detected in the respective method. Thus, the detection rate of intrapulmonary nodules was uninfluenced. In general, no deterioration of image quality was observed. CONCLUSION: On average, 21% dose reduction was achieved by an anatomically adapted tube current modulation in spiral CT examinations of the thorax without a loss in detectability of pulmonary nodules and image quality. This method is an important contribution to dose reduction in spiral CT. PMID- 11414158 TI - [Local therapy of liver metastases of colorectal carcinomas. Laser therapy vs. direct current treatment in the rat model]. AB - PURPOSE: Evaluation of the efficacy of direct current and laser therapy in the treatment of colorectal liver metastases in a rat model. MATERIALS AND METHODS: Liver metastases in 49 rats were induced by subcapsular injection of cells of a colorectal tumor cell line. 3 weeks after inoculation 41 tumors were treated by laser and 56 tumors by direct current therapy. 17 tumors served as a control in the direct current group, 12 in the laser therapy group. The inoculation rate and the tumor size 3 and 5 weeks after therapy were determined by MRI. RESULTS: The MR-tomographically determined inoculation rate was 133 tumors after 151 tumor cell injections (88%). Three and five weeks after laser therapy, tumor growth rates of 15% and 18% were documented (maximum tumor diameter). This represented a significant reduction in growth (p < or = 0.01 and p < or = 0.001) compared to the control group (growth rate 230% and 280%). In the direct current therapy group, growth rates were 40% after 3 and 60% after 5 weeks. Compared to the control group (210% and 290%), this also represented a significant reduction (p < or = 0.001). CONCLUSIONS: This animal study is the first to demonstrate an antitumor effect of direct-current therapy on liver metastases of colorectal cancer in rats. Nevertheless, the laser therapy proved to be more effective. PMID- 11414159 TI - [Proximal inflow obstruction as a rare initial manifestation of mediastinal melanoma metastasis]. PMID- 11414160 TI - [Local thoracic meningocele in scoliosis and neurofibromatosis type I]. PMID- 11414161 TI - [Ingestion of a chicken bone with secondary aspiration]. PMID- 11414162 TI - [Tamoxifen: does a wider range of indications require more careful monitoring?]. AB - Two patients, premenopausal women aged 48 years and 37 years, who were treated with tamoxifen following a mammary carcinoma operation, experienced abdominal complaints, hot flushes, vaginal discharge, an irregular menstrual cycle and/or concern about the increased risk of ovarian carcinoma. Transvaginal ultrasonography of the ovaries and laboratory tests indicated ovarian overstimulation. Both patients temporarily stopped using tamoxifen; in one of the patients both ovaries were ablated. The range of indications for treatment with tamoxifen has in recent years been extended to premenopausal patient groups and the length of treatment has been increased from two to five years. We recommend more extensive checks when prescribing tamoxifen to premenopausal patients in view of the possible adverse effects of ovarian stimulation associated with this treatment. PMID- 11414163 TI - [Safety of specific cyclo-oxygenase 2 inhibitors]. AB - Previous studies have shown that the selective cyclo-oxygenase 2 inhibitors (COXIBs), rofecoxib and celecoxib, were as effective as conventional NSAIDs in relieving both pain and inflammation. In two prospective studies, each including more than 8000 patients, the use of COXIBs was associated with a clinically significant decrease in symptomatic ulcers and their complications (PUBs). In the VIGOR study, the annual incidence of PUBs was 2.1% in the rofecoxib group and 4.5% in the group treated with conventional NSAIDs (relative risk (RR): 0.5; 95% CI: 0.3-0.6). In the CLASS study, comparable results were observed: annual incidence of PUBs was 2.1% in the celecoxib group and 3.5% in the NSAID group (RR: 0.6; 95% CI: 0.4-0.9). In patients using low dose aspirin for cardio protection, there was no difference between celecoxib and NSAIDs with respect to the incidence of PUBs. The use of rofecoxib was associated with a reduction in endoscopies, additional proton pump inhibitors and hospital admissions. In conclusion, COXIBs are a major advance in the prevention of ulcers and their complications for patients who require treatment with NSAIDs. PMID- 11414164 TI - [No indication for thrombophilia screening in patients with idiopathic venous thromboembolism and their relatives]. AB - In 70% to 80% of patients with venous thromboembolism, a thrombophilic defect can be identified. The most important defects are: antibodies to phospholipids or lupus anticoagulants, mutation of factor V Leiden, prothrombin mutation, mild hyperhomocysteinaemia, and increased factor VIII levels. Deficiencies of antithrombin, protein C or protein S are rare. Whether or not screening for thrombophilia in patients with idiopathic venous thromboembolism is justified, depends on the potential benefits for the patients, or their relatives. At present, patients with a thrombophilic defect do not appear to have a much higher risk for recurrent venous thromboembolism, than patients with thrombosis but without a defect. The absolute risks of venous thromboembolism in asymptomatic relatives with a thrombophilic defect are too low to justify initiating a general policy of family screening. In conclusion, a conservative approach towards thrombophilia screening in idiopathic venous thromboembolism is warranted. PMID- 11414165 TI - [Roaming through methodology. XXXIII. Ethics of sample size estimation: less subjects needed for a one-sided than for a two-sided statistical investigation]. AB - In sample size calculations for clinical trials, two-sided statistical testing is the usual starting point. Compared with one-sided testing, this option requires the inclusion of a larger number of study subjects, and a larger number of study events will be needed before a conclusion can be reached. It is therefore ethically relevant to consider in which situations one-sided testing should be the preferred option. One-sided statistical testing is preferable when the research hypothesis is one-sided (is intervention A better and intervention B?), or when only a clearly better result for A compared to B will have clinical consequences (e.g. if A is more cumbersome or more invasive to the patient than B). For the estimation of the number of study subjects needed, one-sided testing should then be assumed. If a new intervention is to be tested by comparison with placebo treatment or with absence of treatment, one-sided testing is an adequate starting point for sample size estimation. PMID- 11414166 TI - [Diagnostic image (40). Scurvy]. AB - In a 17-year-old woman with anorexia nervosa, scurvy was diagnosed as the cause of fatigue, abdominal pain, vaginal bleeding and several distinctive skin lesions. PMID- 11414167 TI - [From gene to disease; achondroplasia and other skeletal dysplasias due to an activating mutation in the fibroblast growth factor]. AB - Achondroplasia, the most common and best known skeletal dysplasia, is inherited in an autosomal dominant fashion. Like a number of other skeletal dysplasias, among which hypochondroplasia and thanatophoric dysplasia, achondroplasia is caused by mutations in the fibroblast growth factor receptor 3 (FGFR3) gene. FGFR3 is a negative regulator of bone growth. Binding of fibroblast growth factors to the FGFR3 receptor stimulates its tyrosine kinase activity in the cell. This activates a signal transduction pathway that regulates enchondral ossification by inhibition of cell division and stimulation of cell maturation and differentiation. Mutations in the FGFR3 gene give rise to activation of the receptor in the absence of growth factors, thus causing abnormal long bone development. Position and type of mutation in the FGFR3 gene determine the extent of overactivation and thus the severity of the skeletal abnormality. PMID- 11414168 TI - [Breast complaints during the Dutch nationwide breast cancer screening program: increased risk of referral and of breast cancer]. AB - OBJECTIVE: To determine the prevalence of breast complaints reported on a questionnaire among participants in the first round of the national breast cancer screening programme in the Netherlands, and to assess the relationship between these complaints and referral and diagnosis of breast cancer. DESIGN: Retrospective controlled study. METHOD: Women in the South screening region of the Dutch nation-wide breast cancer screening programme (Stichting Bevolkingsonderzoek Borstkander Zuid (BoBZ)) received a medical questionnaire together with the invitation for participation in the screening programme. This questionnaire was only reviewed by the radiologist if there were doubts concerning referral. The study assessed whether participants with these complaints had higher risks of referral and screen-detected breast cancer than those without these complaints. In this study two random and independent samples were drawn from the screening files: 1116 women who attended the first screening round and 1113 women who were referred for medical checkup in the same screening round. RESULTS: The prevalence of breast complaints reported in the first screening round was 5.1% (95% CI = 3.9-6.6). More than half of the complaints included pain not related to menses; nipple secretion; changes in skin (retraction or discolouration) or tumours. The referral risk for women with these complaints was more than twice that for women without complaints (adjusted odds ratio (OR) = 2.3; 95% CI = 1.4-3.8) and their risk of screen-detected breast cancer was more than three times as high (adjusted OR = 3.3; 95% CI = 1.4-3.8). Up to 5.8% of the screen-detected breast cancers could be attributed to the breast complaints. CONCLUSION: Breast complaints at the time of breast cancer screening increased the risk of breast cancer being detected, but it is not clear as yet whether the screening will benefit from the explicit incorporation of questions regarding complaints. PMID- 11414169 TI - [High levels of carcinoembryonic antigen in a woman with hypothyroidism]. AB - A 53-year-old woman was referred to the outpatient clinic for Internal Medicine, due to an elevated serum concentration of carcinoembryonic antigen (CEA). Hypothyroidism was diagnosed. Following replacement therapy with levothyroxin, the serum concentrations of thyroid stimulating hormone (TSH) and CEA decreased to near-normal levels. No malignancy was found during the remainder of the treatment period. A decreased hepatic clearance of CEA seemed to be the most likely explanation for the observed high CEA concentration in this patient with hypothyroidism. An increased CEA concentration may occur in association with certain malignancies, as well as with non-neoplastic disorders, such as hypothyroidism. As CEA has a low tumour specificity, routine assessment of CEA is not advocated for diagnostic evaluation. PMID- 11414170 TI - [Doctors are increasingly held liable for missed diagnosis of cancer in mass screening]. AB - Radiologists and pathologists are increasingly subjected to litigation for alleged faults in diagnosing cancer. An important cause of this appears to be the introduction of mass screening for cancer. According to liability law, the judge must assess whether the person making the diagnosis effected his/her duty with due care. The distinction between error and incompetence cannot be judged solely on the basis of the individual case, but requires an estimation of the accuracy of the diagnosis on a population level. The traditional role of the expert witness, who is asked to judge the individual cases, needs to be reconsidered. A re-examination of slides or radiograms with prior knowledge of the outcome obtained does no justice to the original situation and an alternative procedure is therefore needed. Since the government initiates and controls population screening and the professional organisations involved in screening have committed themselves to monitoring the process and the results obtained, the general public must be better informed about the limitations of screening. Unrealistic expectations as to the benefits of screening appear to be an important motive for legal actions. It is questionable whether screening quality can be enhanced by means of the preventive effect of professional liability. Doctors and lawyers should analyse and solve this problem together. PMID- 11414171 TI - [Evaluation report on the Dutch Medical Treatment Act (WGBO)]. PMID- 11414172 TI - [Evaluation report on the Dutch Medical Treatment Act(WGBO)]. PMID- 11414173 TI - [Evaluation of the Dutch Medical Treatment Act(WGBO)]. PMID- 11414174 TI - [Evaluation of the Dutch Medical Treatment Act (WGBO)]. PMID- 11414175 TI - [Responsibility circumvented with a set time for medical record keeping]. PMID- 11414176 TI - [Responsibility circumvented with a set time for medical record keeping]. PMID- 11414177 TI - [Tubercular granulomas of the liver in a Somalian patient with chronic hepatitis C]. PMID- 11414178 TI - Keeping human rights on the bioethics agenda. PMID- 11414179 TI - The integration of health and human rights: an appreciation of Jonathan M. Mann. PMID- 11414180 TI - Bioethics and human rights: a historical perspective. PMID- 11414181 TI - The lingua franca of human rights and the rise of a global bioethic. PMID- 11414182 TI - Beyond moral claims: a human rights approach in mental health. PMID- 11414183 TI - Caretakers and collaborators. PMID- 11414184 TI - A human right to healthcare access: returning to the origins of the patients' rights movement. PMID- 11414185 TI - Proposing a new agenda: bioethics and international human rights. PMID- 11414186 TI - Future animal: environmental and animal welfare perspectives on the genetic engineering of animals. PMID- 11414187 TI - Reconsidering the pseudo-patient study. PMID- 11414189 TI - All that remains: identifying the victims of the Srebrenica Massacre. PMID- 11414190 TI - Why are there no clinical ethicists in France? PMID- 11414191 TI - Toward a historical ethics. PMID- 11414192 TI - Cancer prevention and the contribution of cancer registries. AB - "Because they know what they do" should be the contribution of cancer registration to prevention. The public should be informed about the successes and failures of prevention. In addition, each doctor and each hospital should know the long-term results for its patients despite the complex interdisciplinary health care provision. At the same time, the regional results should be available and contrasted with clinical studies and international standards. An important criterion is also the quality of life of the patients, whose cooperation is more than overdue. According to the possibilities of prevention, six important levels can be differentiated. On each level the outcome should be evaluated on the basis of slightly differing criteria. Primary prevention has the largest incidence-, and thus, mortality-reduction potential, essentially caused by the giving up of smoking. However, the primary prevention of the tumor depends upon what kind of tumor it is, as primary prevention is possible to different extents. Often, secondary prevention is the earliest intervention possible, that is, the early detection of cancer in prognostically favorable stages. However, early detection only offers a chance of cure, albeit a smaller one, for some kinds of tumor. The third prevention level comprises primary care according to the state-of-the-art standards, followed by posttreatment care (disease-free phase) even when the disease is running a fatal course (palliative phase). In the terminal phase, adequate tumor-pain therapy and symptom-oriented measures stand for the sixth prevention level. Even the quality of life of the dying can be optimized. At the beginning of treatment at the latest, the quality of life should also be added to the outcome criteria. In this paper, the individual levels of prevention and possible evaluation criteria for successful prevention, which a modern cancer registration should submit, are discussed and examples given. If the evaluation of the quality of treatment and its significance for health care provision becomes the central tasks of the cancer registers, they will need to be transformed into service centers for hospitals, doctors, and patients. PMID- 11414193 TI - Treatment of chemotherapy-induced nausea and vomiting. PMID- 11414194 TI - Recombinant human erythropoietin increases the radiosensitivity of xenografted human tumours in anaemic nude mice. AB - PURPOSE: The effect of recombinant human erythropoietin (Epo) on the radiosensitivity of human tumour xenografts growing in anaemic nude mice was studied. METHODS AND MATERIALS: Anaemia was induced by total body irradiation (TBI) of mice prior to tumour transplantation. The development of anaemia was prevented by Epo (1000 U/kg s.c.) given 3 times weekly starting 2 weeks prior to TBI (5 Gy). Epo treatment did not influence the growth rate of the tumours, which were transplanted into the subcutis of the hind leg of mice. Thirteen days after TBI (tumour volume of approx. 40 mm3), a single-dose irradiation (12 Gy) of the tumour was performed resulting in a growth delay with subsequent regrowth of the tumours. RESULTS: In Epo-treated animals the tumour growth delay was significantly longer compared to anaemic mice. However, the radiosensitivity of tumours in non-anaemic animals' (non-Epo-treated) tumours could not fully be restored. CONCLUSION: These data give evidence for restored radiosensitivity after correction of anaemia with Epo. PMID- 11414195 TI - Mutations of the p53 gene in malignant rhabdoid tumors of soft tissue and the kidney: immunohistochemical and DNA direct sequencing analysis. AB - Malignant rhabdoid tumor (MRT) is characterized by the presence of intracytoplasmic eosinophilic inclusions composed of whorls of intermediate filaments. This tumor was originally described as an entity of the abortive type of Wilms' tumor in childhood. Recently, it has been proved that these rhabdoid cells can be observed in various types of malignant tumors, including soft tissue sarcoma or carcinoma. To investigate the oncogenesis of this tumor, we examined the p53 gene alteration by means of immunohistochemical analysis and DNA direct sequencing in three cases of malignant rhabdoid tumor (MRT) of the soft tissue and three cases of MRT of the kidney. All the cases of MRT of the soft tissue and two of the cases of MRT of the kidney showed immunopositivity for p53 protein. Among them, one of the cases of MRT of the soft tissue and two of the cases of MRT of the kidney showed missense mutations of the p53 gene. These results strongly suggest that p53 gene alterations may have an important role to play in the aggressive biological behavior and poor prognosis of this tumor. PMID- 11414196 TI - The exogenous wild-type p14ARF gene induces growth arrest and promotes radiosensitivity in human lung cancer cell lines. AB - The cyclin-dependent kinase inhibitor p16INK4a encoded by the INK4A/CDKN2A/MTS1 gene is a frequent target of 9p21 inactivation in human lung cancers. The p14ARF transcript, which is an alternative spliced form of this locus, is also altered or deleted in a proportion of human lung cancers and has been shown to inhibit cell cycle progression as an endogenous cellular regulator of the p53 protein, raising the possibility that it might constitute an additional lung tumor suppressor gene at the 9p21 locus. To test the candidacy of p14ARF as a lung cancer suppressor and assess the role it plays in radiosensitivity, we transfected the wild-type p14ARF gene into four cell lines which had various endogenous gene backgrounds of INK4A-/p53+/RB+ (A549 and H460), INK4A+/p53+/RB- (H446) as well as p14ARF+/p53-/RB+ (Calu-1). We found that transfection of p14ARF is related to an obvious growth inhibition in all wtp53 cell lines, regardless of INK4A/ARF and RB status. Although it has been shown that p53-induced G1 checkpoint in response to DNA damage by ionizing radiation is p14ARF-independent, we found the radiosensitivity of two p14ARF-deficient cell lines was increased after p14ARF gene transfer. The results indicated that cell cycle redistribution after acquiring the exogenous gene might be the main explanation for the enhanced sensitization. An increased radiation-induced apoptotic proportion in one cell line also suggested a fortified p53 function that might be triggered by the restored p14ARF protein. PMID- 11414197 TI - The glutathione-related detoxification system is increased in human breast cancer in correlation with clinical and histopathological features. AB - PURPOSE: The glutathione detoxification pathway includes glutathione S transferase (GST) and peroxidase (GPX) isoenzymes as well as glutathione reductase (GSSR). Though well established from cultured cancer cell lines, its involvement in resistance is still unclear in the tumours. This study aimed to describe the parameters that influence the glutathione contents and associated activities in breast cancer. METHODS: The components of the glutathione pathway were measured in the tumours from 41 women with primary breast cancer in comparison with those in the matched tumour-free samples. Appropriate statistical studies (regression analysis, Wilcoxon signed rank test) explored the influence of clinical and prognostic factors. RESULTS: Reduced and total glutathione contents were largely increased (P < 0.0001) and all related activities were significantly enhanced in the tumours. Interindividual variations were described, probably due to various parameters (age, menopause, axillary lymph node status, S and G2 + M cell fractions, ER, cathepsin-D and c-ErbB-2 expressions) that influence particular components of the glutathione pathway, especially the glutathione levels. CONCLUSIONS: The breast tumours improved their redox status and detoxification capacities depending on various parameters of significance for cell proliferation and aggressiveness, which supports the involvement of the glutathione pathway in malignant cell resistance to oxidative stress and apoptosis. PMID- 11414198 TI - Comprehensive galectin fingerprinting in a panel of 61 human tumor cell lines by RT-PCR and its implications for diagnostic and therapeutic procedures. AB - PURPOSE: Knowledge about galectin expression by human tumor cells is mainly restricted to galectins-1 and -3. This study was conducted to define the gene expression pattern of all presently known human galectins in tumor cell lines of various histogenetic origin (galectinomics). METHODS: The presence of mRNAs for human galectins-1, -2, -3, -4, -7, -8, and -9 was monitored by RT-PCR analyses in a panel of 61 human tumor cell lines of different origin (breast, colon, lung, brain, skin, kidney, urogenital system, hematopoietic system). RESULTS: The validity of the technique was first confirmed by comparison of RT-PCR data with those obtained by Western blotting and cytofluorometry for galectins-1 and -3 in 18 cell lines. The following detection of a complex pattern of gene expression beyond commonly studied galectins-1 and -3 underscored the need for this fingerprinting. The most abundantly expressed message for a member of this lectin family was galectin-8 with 59 positive cell lines. With the exception of the tested lung tumors, galectin-1 and -3 transcripts were frequently expressed in the cell line panel with differences between individual cases. Positivity for galectins-2 and -4 was confined to a significant fraction of colorectal and neural tumors. Signals for galectin-9, the third known human tandem-repeat-type galectin besides -4 and -8, appeared in colorectal carcinoma cell lines with a frequency similar to that of galectin-4 but with inter-line differences. Its expression was restricted to lines of this tumor type, of the tested ovarian carcinoma, and hematopoietic malignancies. CONCLUSIONS: The results clearly demonstrate that human tumor cells express more mRNA species for galectins than those for galectins-1 and -3. To derive unequivocal diagnostic and prognostic information by immunohistochemistry on galectins with antagonistic impact on growth control and significant influence on cell adhesion, additional monitoring of these so far insufficiently studied family members is essential. PMID- 11414199 TI - Salvage chemotherapy with mitoxantrone, fludarabine, cytarabine, and cisplatin (MIFAP) in relapsing and refractory lymphoma. AB - PURPOSE: The aim of the study was to evaluate the feasibility and efficacy of the combination of mitoxantrone, fludarabine, cytarabine, and cisplatin (MIFAP) in patients with prognostically unfavorable recurrent and refractory Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL). METHODS: Forty-six patients (median age 43 years, range 18-63) with relapsed (n = 15) or refractory (n = 31) malignant lymphoma were enrolled (HD, n = 13; low-grade/transformed NHL, n = 4; high-grade NHL, n = 29). A total of 39 patients (85%) showed multiply relapsed diseases with a duration of prior remission of < 12 months (n = 8) or had lymphoma being resistant to prior chemotherapy (n = 31). The MIFAP therapy consisted of fludarabine (15 mg/m2, q. 12 h, day 1-4), cytarabine (50 mg/m2 by continuous infusion (CI) over 22 h, day 1-4), cisplatin (25 or 30 mg/m2 by CI over 24 h, day 1-4), and mitoxantrone (4 mg/m2, day 2-5). RESULTS: Thirteen patients (28%) achieved complete remission (CR) and 15 patients (33%) partial remission (PR), for an overall response (OR) rate of 61%. Twenty-two patients responding to MIFAP (10 CR, 12 PR) have been consolidated by high-dose therapy (HDT) with hematopoietic stem cell transplantation (SCT). After a median follow up of 12 months, 16 patients are in continuous CR (CCR) (n = 14) or CCRu (unconfirmed) (n = 2). The median duration of event-free survival (EFS) and overall survival (OS) were 6.5 and 19.3 months, respectively. Probabilities of EFS and OS after 3 years were 19% and 40%. Responders consolidated by subsequent HDT showed rates for 3-year EFS and OS of 40% and 66%, respectively. Unfavorable prognostic factors for EFS by univariate analysis were refractory lymphoma and the presence of B-symptoms. Significant prognostic factors for OS were NHL, refractory lymphoma, B-symptoms, and bone marrow involvement. The major toxicities were leukocytopenia and thrombocytopenia of the World Health Organization (WHO) grade IV in nearly all courses (median duration 10 and 11 days). In contrast, non-hematological side effects were moderate, predominantly of WHO grades I and II. Treatment-related mortality with MIFAP was 4% (two patients with septicemia by Aspergillus fumigatus). CONCLUSIONS: MIFAP is an effective salvage protocol for patients with poor-risk recurrent or refractory HD and NHL. The observed toxicity seems to be acceptable considering the unfavorable prognosis and intensive pretreatment. The results in patients responding to MIFAP and afterwards undergoing HDT with autologous stem cell support are even comparable to those published in patients with prognostically more favorable diseases. PMID- 11414200 TI - Angiosarcoma of the scalp: treatment with liposomal doxorubicin and radiotherapy. AB - PURPOSE: Angiosarcoma of the scalp and face is a rare malignant endothelial tumor arising mainly in elderly people. The prognosis is poor. Effective and safe treatments are warranted. METHODS: A 79-year-old woman with an angiosarcoma of the scalp larger than 5 cm in diameter was treated with intravenous liposomal doxorubicin, 20 mg per square meter body surface (i.e., 30 mg) once per month followed by radiotherapy. RESULTS: After 12 infusions of liposomal doxorubicin, we observed a partial remission with a > 50% decrease of affected area and disappearance of ulceration. After 21 infusions, however, there was no further improvement. We decided to discontinue chemotherapy but move on with radiotherapy with an electron beam using fractionated doses of 2 Gy five times per week for up to a total of 40 Gy. To ensure a maximum dose in the upper layer of the dermis a bolus technique was used. Radiotherapy was terminated due to a temporary circumscribed epidermolysis. At the end of treatment a remarkable regression of the cutaneous lesion was noted. During the subsequent 24 months she has not developed any metastatic spread. CONCLUSION: Sequential therapy of bad prognosis angiosarcoma with liposomal doxorubicin followed by radiotherapy showed a marked clinical improvement and prolonged relapse-free survival in this patient. PMID- 11414201 TI - The importance of reliable exposure estimates in deciding whether trichloroethylene can cause kidney cancer. PMID- 11414203 TI - MBP fusion protein with a viral protease cleavage site: one-step cleavage/purification of insoluble proteins. PMID- 11414204 TI - Sensitive ribonuclease protection assay employing glycogen as a carrier and a single inactivation/precipitation step. PMID- 11414205 TI - Modified version of pACT-2 that simplifies cloning with NdeI. PMID- 11414206 TI - Vector for high-throughput sequencing: construction and preparation with cyclic cut-ligation. PMID- 11414207 TI - Generation of baculovirus recombinants using PCR-amplified fragments. PMID- 11414208 TI - Hybridization to high-density filter arrays of a Brugia malayi BAC library with biotinylated oligonucleotides and PCR products. PMID- 11414209 TI - Enhanced capture of small histidine-containing polypeptides on membranes in the presence of ZnCl2. PMID- 11414210 TI - Sending plasmid DNA by mail. PMID- 11414211 TI - Preparation of mitochondrial DNA from yeast and filamentous fungi. PMID- 11414212 TI - Multicolor imaging: the important question of co-localization. PMID- 11414213 TI - Macro for analysis of CpG and CpNpG methylation in plants. PMID- 11414214 TI - Cloning full-length, cap-trapper-selected cDNAs by using the single-strand linker ligation method. AB - We have developed the single-strand linker ligation method (SSLLM), which uses DNA ligase to add a dsDNA linker to single-stranded (ss) full-length cDNA. The linkers have random 6-bp (dN6 or dGN5) 3' overhangs that can ligate to any cDNA sequence, thereby facilitating the production of cDNA libraries with titers exceeding 1 x 10(6) independent clones. We confirmed that the 5' ends of cDNA inserts cloned by using SSLLM are full-length and include the 5' untranslated regions. The great advantage of our method is that the elimination of the GC tail simplifies the sequencing and protein translation of the full-length clones. Further, our method tags ss cDNAs more efficiently than does the traditional RNA ligase reaction. PMID- 11414215 TI - RT-PCR method to quantify vascular endothelial growth factor expression. AB - The vascular endothelial growth factor (VEGF) is implicated in the progression of cancers. Its expression is well correlated with tumor growth and metastases. The availability of a rapid and sensitive method to detect the amounts of VEGF mRNA in biological samples of limited size, very small biopsies, or samples containing relatively few cells could provide an interesting prognostic tool for clinicians. We have developed an RT-PCR method that allows us to detect the VEGF mRNA from as little as 3 micrograms total mRNA. We have also shown that this protocol can be generalized to all cell lines tested. This method constitutes a very potent tool for the analysis of VEGF mRNA expression in different contexts. PMID- 11414216 TI - Use of multiple primers in RAPD analysis of clonal organisms provides limited improvement in discrimination. AB - Randomly amplified polymorphic DNA (RAPD) analysis using two or more primers has been reported to provide additional discriminatory ability over one primer used individually. This may be of particular application in epidemiological typing of clonal organisms, such as Shiga toxin-producing E. coli O157, where strain differentiation can be difficult. Using four arbitrary primers individually, and in all possible permutations, E. coli O157 isolates and other arbitrarily chosen E. coli strains were typed using RAPD analysis. For most nonclonal strains, the use of two primers resulted in increased differentiation between isolates; however, more than two primers did not increase further the discriminatory capacity. E. coli O157 isolates that produced virtually identical profiles using one primer did not show increased differentiation when using two or more primers, demonstrating that in some cases, where strains of an organism are highly related, there is limited advantage to using more than one primer in RAPD analysis. PMID- 11414217 TI - Novel signal amplification technology with applications in DNA and protein detection systems. AB - A non-enzymatic approach to signal amplification has practical advantages over conventional target amplification methods. We have designed a simple, cost efficient signal amplification system that can be used to enhance the detection of nucleic acids or protein. The signal amplification process requires initial capture of analyte by a specific probe, which, depending on the analyte, can be an oligomer or an antibody. Once the analyte is captured, amplification moieties are applied to significantly enhance the sensitivity of analyte detection. Nucleic acid amplification is typically greater than 1000-fold, increasing the sensitivity of target detection to less than 1 amol/100 microL. This amplification strategy presents a very flexible system with components that are easily altered to accommodate diverse assay requirements. PMID- 11414218 TI - Fluorescent staining of glycoproteins on polyvinylidene difluoride membrane with 8-aminonaphthalene-1,3,6-trisulfonate. AB - Here, we describe a simple and sensitive method that allows fluorescent detection of glycoproteins on polyvinylidene difluoride (PVDF) membrane. We used periodic acid oxidation of carbohydrate chains of glycoproteins and fluorescent labeling with 8-aminonaphthalene-1,3,6-trisulfonate (AN-TS) by reductive amination. We developed an additional method to enhance the ability of PVDF to absorb glycoproteins by using non-glycoprotein lectin, such as wheat germ agglutinin (WGA), as a link between the PVDF membrane and glycoproteins, resulting in considerably increased detection sensitivity to glycoproteins. PMID- 11414219 TI - Improved technique for detection of enhanced green fluorescent protein in transgenic mice. AB - One of the most exciting recent advances in cell biology is the possibility to use the green fluorescent protein and its various mutated forms as reporter proteins in studies carried out in vitro and in vivo. In the present study, several detection techniques for the enhanced green fluorescent protein (EGFP) were compared in transgenic mice, using fluorescence and confocal microscopy. In addition, different tissue preparation techniques (squash preparations, vibratome sections, frozen sections) were evaluated. As a model we used transgenic mice expressing EGFP under the control of a 5.0-kb fragment of the glutathione peroxidase isoenzyme 5 protein promoter (GPX5-EGFP) or under a 3.8-kb fragment of the cysteine rich protein-1 promoter (CRISP1-EGFP). In the GPX5-EGFP mice, expression of EGFP was observed in the distal part of the caput epididymis, while the CRISP1 promoter directed EGFP expression in the tubular compartment of the testis. Among the various tissue preparation procedures tested, the best morphological and histological preservation, and reproducibility in EGFP detection, were obtained using frozen sections after a slow tissue-freezing protocol developed in the present study. After slow tissue freezing, specimens of testis and epididymis could be stored at -70 degrees C for at least six weeks without any affect on EGFP fluorescence. Hence, the method developed offers the possibility to analyze EGFP fluorescence in tissues several weeks after specimen collection. The sensitivity achieved was equal to that found in immunohistochemistry, applying biotin-streptavidin-FITC detection. Confocal microscopy is known to have the advantage that fluorescence can be detected from cells in different layers. This was found to be important as regards detecting EGFP fluorescence because the fluorescence was destroyed at the cut surfaces of sections produced by either vibratome or cryomicrotome. PMID- 11414220 TI - ROCK: a spreadsheet-based program for the generation and analysis of random oligonucleotide primers used in PCR. AB - Oligonucleotide primers used to amplify target DNA regions via PCR should meet certain design criteria to maximize the potential for efficient priming. The Random Oligonucleotide Construction Kit (ROCK), a spreadsheet-based program that runs under Microsoft Excel 97 or later version for Microsoft Windows, was developed to facilitate the design of efficient random oligonucleotide primers. Primer sequences are generated that meet user-defined criteria with regard to G + C content, size of a 3' GC clamp, maximum intramolecular/intermolecular complementation potential, and maximum intersequence similarity. The user can analyze the intramolecular/intermolecular complementation potential of program generated primer sequences or of sequences entered manually. The latter may contain any of the standard nucleotide symbols, including ambiguous bases. Primer sequence length, GC%, individual base composition, molecular weight, approximate melting temperature, and mass/volume/concentration relationships can be determined for any sequence generated by ROCK or entered manually. PMID- 11414221 TI - RepGene: a spreadsheet template for the management of reporter gene assays. AB - One of the most powerful techniques in molecular biology is the controlled expression of specific proteins by transfection of eukaryotic cells. This method has become feasible and highly sensitive and, thus, suitable for high-throughput reporter gene assays in basic and applied research. Moreover, the limiting factors are neither the transfection efficiency nor the functional analysis, but rather the ability to manage complex experimental protocols when multiple genes are co-transfected and/or when the effects of several chemical compounds are investigated within the same experiment. Here, we describe an easy-to-use and highly flexible spreadsheet template intended to rationalize and expedite the organization and data management of multi-step reporter gene assays. The objectives of this spreadsheet template are the design of the transfection protocol, the coordination of the administration of test compounds, and the graphical presentation and statistical analysis of the results. PMID- 11414222 TI - Software scripts for quality checking of high-throughput nucleic acid sequencers. AB - We have developed a graphical interface to allow the researcher to view and assess the quality of sequencing results using a series of program scripts developed to process data generated by automated sequencers. The scripts are written in Perl programming language and are executable under the cgibin directory of a Web server environment. The scripts direct nucleic acid sequencing trace file data output from automated sequencers to be analyzed by the phred molecular biology program and are displayed as graphical hypertext mark-up language (HTML) pages. The scripts are mainly designed to handle 96-well microtiter dish samples, but the scripts are also able to read data from 384-well microtiter dishes 96 samples at a time. The scripts may be customized for different laboratory environments and computer configurations. Web links to the sources and discussion page are provided. PMID- 11414223 TI - Replacement of an old HPLC integrator with an inexpensive computer and custom software. PMID- 11414224 TI - LabVelocity: online tools for life science products, protocols, technical information, MEDLINE searches, and laboratory calculations. AB - As the pace of life science discovery increases, so do the demands on researchers. To remain competitive in the life science industry, researchers must use every tool at their disposal to keep up with new products, protocols, news, and literature in their field. While there are now myriad Web sites that assist researchers with this problem, many suffer from confusing user interfaces, poorly designed search engines, and a narrow information focus. Here, we present LabVelocity, a user-friendly Web site that provides a free multidisciplinary information-gathering service for the life science research community. Using LabVelocity, a researcher can quickly find the products, protocols, technical references, news, MEDLINE abstracts, and interactive software tools necessary for an experiment. This aggregation of information can streamline experimental planning and is especially useful when researchers want to set up a new laboratory or to venture outside their field of expertise. PMID- 11414225 TI - Electrophoresis gel image processing and analysis using the KODAK 1D software. AB - The present article reports on the performance of the KODAK 1D Image Analysis Software for the acquisition of information from electrophoresis experiments and highlights the utility of several mathematical functions for subsequent image processing, analysis, and presentation. Digital images of Coomassie-stained polyacrylamide protein gels containing molecular weight standards and ethidium bromide stained agarose gels containing DNA mass standards are acquired using the KODAK Electrophoresis Documentation and Analysis System 290 (EDAS 290). The KODAK 1D software is used to optimize lane and band identification using features such as isomolecular weight lines. Mathematical functions for mass standard representation are presented, and two methods for estimation of unknown band mass are compared. Given the progressive transition of electrophoresis data acquisition and daily reporting in peer-reviewed journals to digital formats ranging from 8-bit systems such as EDAS 290 to more expensive 16-bit systems, the utility of algorithms such as Gaussian modeling, which can correct geometric aberrations such as clipping due to signal saturation common at lower bit depth levels, is discussed. Finally, image-processing tools that can facilitate image preparation for presentation are demonstrated. PMID- 11414226 TI - Biological methods for cell-cycle synchronization of mammalian cells. AB - Understanding the molecular and biochemical basis of cellular growth and division involves the investigation of regulatory events that most often occur in a cell cycle phase-dependent fashion. Studies examining cell-cycle regulatory mechanisms and progression invariably require cell-cycle synchronization of cell populations. Thus, many methods have been established to synchronize cells at specific phases of the cell cycle. Several of the common methods involve pharmacological agents, which act at various points throughout the cell cycle. Because of adverse cellular perturbations resulting from many of the synchronizing drugs used, other synchrony methods that involve less perturbation of biological systems, such as serum deprivation, contact inhibition, and centrifugal elutriation have a significant advantage. The advantages and disadvantages of these cell synchronization methods are discussed in this review. PMID- 11414227 TI - Multiplex capillary denaturing high-performance liquid chromatography with laser induced fluorescence detection. AB - Denaturing high-performance liquid chromatography (DHPLC) is a sensitive, robust, and operationally inexpensive method for the detection of single-base substitutions and small deletions and insertions. To increase sample throughout, we have developed a multiplexing strategy using fluorophores to distinguish different PCR products. The system combines recent advances in the synthesis of monolithic poly(styrene-divinylbenzene) capillary columns with four-color confocal argon ion laser-induced fluorescence detection. Depending on the change in retention caused by the fluorophores, adjustments in the analysis temperature may be required to ensure the maximum mutation detection sensitivity. PMID- 11414228 TI - Silver ion microplates for immunoassays. AB - Microplate wells can be coated with silver ions using glutaraldehyde as a spacer molecule and thiourea as a complexing ligand. Microwells containing surface silver ions are shown to immobilize biotin-labeled horseradish peroxidase (HRP) in active form, while showing very little affinity for the unlabeled enzyme. These plates can also immobilize biotin-labeled antibodies that exhibit bioactivity after immobilization. Silver ions are needed for the complexation of the biotinylated enzyme or antibody because microwells modified to contain surface amine or thiourea molecules do not immobilize appreciable amounts of the labeled proteins. A maximum surface coverage for biotin-labeled HRP of 40 ng/cm2 and an immobilization binding constant of Km = 8 x 10(9)/M are determined from serial dilutions in a microplate. Detection of as little as 6.7 fmol HRP is achieved using antibodies immobilized on the silver ion-modified microplates. Active antibody surface densities were estimated to be between 130 and 260 nm2/antibody molecule. Background binding of HRP to the modified silver ion microplates was very low, allowing for reasonably accurate detection between 10( 14) and 10(-11) mol HRP. PMID- 11414229 TI - Reliability of mRNA profiling: verification for samples with different complexities. AB - Normalization of mRNA profiling data remains an open issue, which turns critical when comparing divergent samples or mRNA populations with different complexities. To address this question, we generated samples with different RNA amounts and complexities by subcellular fractionation of cytoplasmic RNA into the mutually exclusive ribosome-free and polysome-bound RNA pools. For each of the 563 mRNAs analyzed, the hybridization signal corresponding to the cytoplasmic sample equals the sum of signals from the ribosome-free plus the polysome-bound targets (cytoplasmic mRNA = ribosome-free mRNA + polysome-bound mRNA). This intuitive equation was fulfilled only after data normalization following "spiking" of the samples with an exogenous RNA. This is the first demonstration that spiking allows one to correct not only for differences in reaction efficiencies but also to reflect the variations in amount and complexity between the initial mRNA populations. PMID- 11414230 TI - Small-sample total RNA purification: laser capture microdissection and cultured cell applications. AB - Gene expression studies require analysis of RNA, but isolation of total RNA from very small samples by traditional methods can be difficult and inefficient. The Absolutely RNA microprep kit provides a convenient method for isolating total RNA from small numbers of cells such as those harvested by laser capture microdissection (LCM). The protocol includes binding of RNA to a solid support, thus eliminating the need for organic extraction and alcohol precipitation. DNase digestion on the solid support reduces or eliminates DNA contamination and minimizes RNA handling. Efficient washing removes contaminants, and elution in a small volume of buffer results in high-purity RNA at a concentration appropriate for demanding applications such as RT-PCR. RNA isolated from as few as 200 laser capture microdissected brain tumor cells resulted in detection of low, medium, and highly expressed genes by conventional and real-time RT-PCR. PMID- 11414231 TI - JMM, past and present. PMID- 11414232 TI - Belgian Society of Fundamental and Clinical Physiology and Pharmacology autumn meeting. 18 November 2000, Leuven, Belgium. Abstracts. PMID- 11414233 TI - Proceedings of the 1st International Conference on Chemistry and Biology of Vitamin D Analogs. Providence, Rhodes Island, USA. September 26-28, 1999. PMID- 11414234 TI - Lipid-lowering therapy in diabetes mellitus. AB - Dyslipidemia emerges as an important modifiable risk factor for cardiovascular disease in diabetes mellitus, especially as part of the metabolic syndrome in type 2 diabetes. In type 1 diabetes mellitus, tight glucose regulation usually will correct dyslipidemia. Both total cholesterol and triglyceride levels predict cardiovascular disease in diabetes, and HDL-cholesterol may prove to be an even better predictor. In type 2 diabetes, increased triglyceride and reduced HDL cholesterol levels are the key characteristics of dyslipidemia. Increased hepatic VLDL production and impaired catabolism of triglyceride-rich particles contribute to hypertriglyceridemia. Subsequent formation of small dense LDL particles leads to increased atherogenicity. Small dense LDL particles have a longer circulation time, are susceptible to glycoxidation, and are taken up by macrophages and the vessel wall. Post-hoc analysis of diabetic subgroups in primary and secondary prevention trials suggest that individuals with diabetes may enjoy substantial cardiovascular risk reduction from lipid-lowering therapy. Trials prospectively addressing the benefit of lipid lowering therapy in diabetes are under way. Target levels for lipid lowering therapy in diabetes at present stem from pathophysiological plausibility rather than from clinical proof. Intensive lipid lowering with a statin in adequate dosage or a combination of a statin and a fibrate may be used to lower LDL-cholesterol levels to values < 2.6 mmol/l and triglyceride levels to < 1.7 mmol/l, a value at which few small dense LDL particles remain in circulation. Effective medication to raise HDL-cholesterol levels adequately are not yet available for clinical use. Treatment of diabetic dyslipidemia should be as simple as possible, given the polypharmacy that is often necessary for the patient with diabetes. Therefore, single treatment with a statin in adequate dosage is the first choice. PMID- 11414235 TI - Chylothorax complicating chronic lymphocytic leukemia. PMID- 11414236 TI - Photo quiz. Congenital cutaneous candidiasis. PMID- 11414238 TI - Anaerobic Digestion of Solid Waste II. Symposium proceedings. Barcelona, Spain, June 1999. PMID- 11414237 TI - [Therapeutic approach based on evidence]. PMID- 11414239 TI - Risk of advanced proximal neoplasms in asymptomatic adults according to the distal colorectal findings. PMID- 11414240 TI - Preoperative versus postoperative endoscopic retrograde cholangiopancreatography in mild to moderate pancreatitis: a prospective randomized trial. PMID- 11414241 TI - Endosonographic features predictive of benign and malignant gastrointestinal stromal cell tumors. PMID- 11414242 TI - LightCycler-based quantitative PCR for detection of cytomegalovirus in blood, urine, and respiratory samples. PMID- 11414243 TI - Likelihood ratios for microbiology. PMID- 11414244 TI - Improved primer design for multiplex PCR analysis of vancomycin-resistant Enterococcus spp. PMID- 11414245 TI - Clarifications regarding the 3' repeat region of the cagA gene in Helicobacter pylori and clinical outcome. PMID- 11414246 TI - Molecular diagnosis of an Enterocytozoon bieneusi human genotype C infection in a moderately immunosuppressed human immunodeficiency virus seronegative liver transplant recipient with severe chronic diarrhea. PMID- 11414247 TI - Dual emergence in food and humans of a novel multiresistant serotype of Salmonella in Senegal: Salmonella enterica subsp. enterica serotype 35:c:1,2. PMID- 11414248 TI - Pooling cervical swabs for detection of Chlamydia trachomatis by PCR: sensitivity, dilution, inhibition, and cost-saving aspects. PMID- 11414249 TI - The quantitative risks of mesothelioma and lung cancer in relation to asbestos exposure. PMID- 11414250 TI - Asbestos and cancer. PMID- 11414251 TI - Asbestos and cancer risk. PMID- 11414252 TI - N-terminal truncated amyloid beta-peptides and Alzheimer's disease. PMID- 11414253 TI - Advances in emulsion science for personal care applications. PMID- 11414254 TI - Beyond rheology modifications: hydrophilically modified silicone elastomers provide new benefits. PMID- 11414255 TI - The importance of mesomorphic (lamellar) phases in emulsion stability. PMID- 11414256 TI - Self-emulsification of surfactant-oil mixtures produced by diffusion and chemical reaction. AB - Drops having diameters of order 100 microm and containing various combinations of oils, surfactants, and in some cases alcohols were injected into water or aqueous salt or buffer solutions. The resulting dynamic behavior was observed by videomicroscopy. Spontaneous emulsification yielding oil droplets a few microns in diameter was seen in a variety of systems when diffusion and/or chemical reaction caused inversion of the drop from an oil-continuous to a water continuous phase, leading to local supersaturation in oil. Surfactants used included nonionic (C12E6), anionic (Aerosol-OT), and zwitterionic (tetradecyldimethylamine oxide). In some experiments inversion occurred because a lipophilic surfactant was converted to a hydrophilic surfactant, e.g., a double chain phospholipid to two single-chain surfactants. PMID- 11414257 TI - Calciotropic hormones and the skin: a millennium perspective. PMID- 11414258 TI - Meadowfoam derivatives: technologically advanced cosmetic ingredients. PMID- 11414259 TI - Structure/activity relationship in cosmetic raw material: tailor-made natural blends. PMID- 11414261 TI - Proceedings of the 9th International Symposium on Subtypes of Muscarinic Receptors. October 31-November 4, 2000. Houston, Texas, USA. PMID- 11414260 TI - Surfactin sodium salt: an excellent bio-surfactant for cosmetics. PMID- 11414262 TI - Antihypertensive medication use in Hispanic adults: a comparison with black adults and white adults. AB - BACKGROUND: Variations in awareness, treatment, and control of hypertension among different racial/ethnic groups have been widely reported. It is unclear whether these differences are explained fully by differences in socioeconomic status, insurance coverage, health status, and health behaviors, or whether these differences indicate that racial/ethnic subgroups have unique barriers to hypertension control. OBJECTIVES: Determine whether there are significant differences between racial/ethnic groups in medication use for hypertension after adjusting for potentially confounding variables. RESEARCH DESIGN: Cross-sectional analysis of the 1992 Health and Retirement Study. SUBJECTS: 2450 non-Hispanic white, 939 non-Hispanic black, and 345 Hispanic participants, ages 51 to 61, reporting a history of hypertension. MEASURES: Self-reported current antihypertensive medication use. We used logistic regression to adjust for demographics, socioeconomic status, health status, insurance, and health risk behaviors. RESULTS: 63.6% of white adults, 72.6% of black adults, and 52.5% of Hispanic adults reported current medication use to control hypertension (P <0.001 across all three groups). In stratified analysis, the lower rate of use for Hispanic adults was consistent regardless of gender, insurance coverage, or health status. After controlling for all variables, the adjusted prevalence for Hispanic adults was 50.8% and 73.3% for black adults. CONCLUSIONS: The differences in antihypertensive medication use between white adults, black adults, and Hispanic adults, particularly the markedly lower rates among Hispanic adults, are not explained by differences in demographics, socioeconomic status, health insurance coverage, health status, or health risk behaviors. Alternative explanations for these results and areas for future research and intervention are explored. PMID- 11414263 TI - Administration gives "OK" to privacy regulations. PMID- 11414264 TI - Variable effects of social support by race, economic status, and disease activity in systemic lupus erythematosus. AB - OBJECTIVE: Health status and disease activity in patients with systemic lupus erythematosus (SLE) and other chronic diseases are strongly associated with social support, which suggests that enhanced social support in medical care might improve SLE outcome. There has been little or no study on identifying patients for whom social support would be most beneficial. It would allow practitioners to enable social support more effectively as a complement to disease management. METHODS: A retrospective cohort (200 patients with SLE from 5 centers), balanced by race and insurance status, was studied in a cross sectional design. Demographic, clinical, socioeconomic, and psychosocial factors and disease outcomes [Systemic Lupus Activity Measure (SLAM), Systemic Lupus International Collaborating Clinics/American College of Rheumatology (SLICC/ACR), SF-36] were measured. Using multivariate linear regression and ANOVA (outcome = SF-36 physical function, mental health), we examined the interaction between social support and patients' characteristics, including race, age, income, occupation, insurance, employment, education, and social network, and characteristics of the disease itself such as disease activity (SLAM) at diagnosis, damage (SLICC/ACR) at diagnosis, and comorbidity. RESULTS: In multivariate models, higher social support was significantly associated (p < 0.05) with better physical function when respondents were white, had income above poverty level, had Medicare or private insurance (vs Medicaid or no insurance), and had low disease activity at diagnosis. Social support was associated with better mental health, although there was no significant interaction between social support and other predictors of mental health. CONCLUSION: The data suggest that social support is beneficial for mental health for all groups, but has the greatest opportunity for influence among those already possessing social, economic, and health advantages. PMID- 11414265 TI - Nomenclature for factors of the HLA system, update March 2001. PMID- 11414266 TI - Amycolatopsis rubida sp. nov., a new Amycolatopsis species from soil. AB - The taxonomic position of a soil isolate, strain 13.4T, was established using a polyphasic approach. The organism was found to have chemical and morphological properties consistent with its classification in the genus Amycolatopsis. Phylogenetic analysis of the strain based on its 16S rDNA sequence showed that it forms a distinct phyletic line within members of the genus Amycolatopsis. The organism was also readily distinguished from the type strains of all validly described Amycolatopsis species by its phenotypic features. The name Amycolatopsis rubida sp. nov. is proposed for this new species. The type strain is strain 13.4T (= AS 4.1541T = JCM 10871T). PMID- 11414267 TI - Phylogenetic relationships of Anaplasma marginale and 'Ehrlichia platys' to other Ehrlichia species determined by GroEL amino acid sequences. AB - The heat-shock protein (GroEL) genes of Anaplasma marginale, Ehrlichia muris and 'Ehrlichia platys' were sequenced and compared with the GroEL of other species of Ehrlichia. The GroEL amino acid sequences of A. marginale and 'E. platys' were most similar to the GroEL sequence of Ehrlichia phagocytophila, with which they formed one group with 6-10% divergence. The E. muris GroEL was most closely related to the GroEL of two unclassified strains (HF-565 and Anan), then to Ehrlichia chaffeensis, Ehrlichia canis, Ehrlichia ewingii and Cowdria ruminantium, forming a second distinct group (0.3-8.6% divergence). The GroELs of Ehrlichia risticii and Ehrlichia sennetsu were very similar to one another (only 2% divergence), forming the third group. The first two groups were relatively closely related (17-20% divergence), while the third group was only distantly related to the first two groups (62-73% divergence). PMID- 11414268 TI - Alkalibacterium olivoapovliticus gen. nov., sp. nov., a new obligately alkaliphilic bacterium isolated from edible-olive wash-waters. AB - A novel Gram-positive, obligately alkaliphilic, non-sporulating, rod-shaped, flagellated bacterium is described. Three different strains of the bacterium were isolated from the wash-waters of edible-olive production. The strains are motile, psychrotolerant, halotolerant, facultatively anaerobic bacteria with a pH optimum of 9.0-9.4 for two strains and 9.8-10.2 for the third. They are catalase- and oxidase-negative. A range of hexoses and some disaccharides composed of hexoses, but not pentoses are metabolized by the bacterial strains: D(+)-glucose, D(+) glucose 6-phosphate, D(+)-cellobiose, starch or sucrose are the carbohydrates best utilized. No common amino acids are utilized by the three alkaliphilic strains, but yeast extract can serve as sole carbon and energy source. The major membrane phospholipids are diphosphatidylglycerol, phosphatidylglycerol and an unknown phospholipid, all containing saturated and unsaturated, even-carbon numbered fatty acyl chains with hexadecanoic and hexadecen(7)oic as the predominant components. The G+C content of the DNA in all three strains is 39.7+/ 1.0 mol% and the DNA relatedness by hybridization is >88% for all pairings of the three strains. The results of 16S rRNA sequence comparisons revealed that the strains represent a new alkaliphilic linkage in the order Bacillales, belonging to the Carnobacterium/Aerococcus-like spectrum. It is proposed that the strains should be assigned to a new genus and species, Alkalibacterium olivoapovliticus. The three strains, designated WW2-SN4aT, WW2-SN4c and WW2-SN5, have been deposited with Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) as DSM 13175T, DSM 12937 and DSM 12938 respectively, and in the National Collection of Industrial and Marine Bacteria as NCIMB 13710T, NCIMB 13711 and NCIMB 13712, respectively. The type species of this genus is Alkalibacterium olivoapovliticus and the type strain is WW2-SN4aT. PMID- 11414269 TI - Circadian differences among 4,096 emergency department patients with acute asthma. AB - INTRODUCTION: In patients with asthma, airways narrow during the night. The clinical implications of a nocturnal presentation of patients with acute asthma to the emergency department (ED) are uncertain. OBJECTIVE: Our objective was to determine whether patients with asthma who had ED visits during the night (midnight to 7:59 am) vs. other times were more severe, responded less well to ED therapy, and had worse clinical outcomes. DESIGN AND SETTING: We performed a cohort study, as part of the Multicenter Airway Research Collaboration (n = 77 sites). ED patients with acute asthma, ages 2-54 yrs, underwent a structured interview in the ED. Chart review of missed/refusal patients created a truly consecutive case series. MEASUREMENTS AND MAIN RESULTS: Among 1,602 children, 19% presented at night Nighttime patients were more likely to be younger, male, and have a shorter duration of symptoms; there were no other clinical differences noted. Among 2,494 adults, 20% presented at night, and they were more likely to be female and to have a history of steroid use for asthma. Nighttime adults also had a shorter duration of symptoms and slightly lower peak flows (mean, 45% vs. 49% of predicted; p = .006) and were more likely to receive steroids. They were more likely to be intubated (2.0% vs. 0.2%; p < .001), but, overall, they were equally likely to be admitted or relapse after ED discharge. In contrast to objective measures of acute asthma severity, both nighttime children and adults were significantly less likely to report their asthma symptoms as severe. CONCLUSION: Except for endotracheal intubation (in adults only), circadian differences minimally affect ED presentation, therapy, or the outcomes of acute asthma. Nighttime asthmatics may be relatively insensitive to the symptoms of severe asthma. PMID- 11414270 TI - Succinylcholine-induced hyperkalemia in a patient with mucositis secondary to chemotherapy. AB - OBJECTIVE: To report a case of fatal hyperkalemia owing to succinylcholine administration in a patient with mucositis secondary to chemotherapy. DESIGN: Case report. SETTING: Adult intensive care unit (ICU) at Dartmouth-Hitchcock Medical Center. PATIENTS: One patient with mucositis secondary to chemotherapy. MEASUREMENTS AND MAIN RESULTS: A 37-yr-old female with recently diagnosed acute myelogenous leukemia was admitted to the ICU with mental status changes and progressive dyspnea requiring intubation and mechanical ventilation. Before ICU admission, the patient had suffered from painful mucositis causing severe dysphagia and bleeding, which was thought to be the result of chemotherapy. By the 10th ICU day, the patient's respiratory and mental status improved and the patient was successfully extubated. However, 8 hrs after extubation, she gradually developed severe respiratory distress, requiring reintubation and mechanical ventilation. The patient was given 14 mg of etomidate and 100 mg of succinylcholine intravenously. Immediately after the intubation, she suffered cardiac arrest. Her serum potassium level was 13.1 mEq/L and HCO3- was 16 mEq/L. The resuscitation attempt was unsuccessful, and the patient was pronounced dead. CONCLUSION: Oral mucositis is a frequent and potentially severe complication of cancer chemotherapy. We believe that mucositis was a contributing factor to this case of fatal hyperkalemia after administration of succinylcholine, with a mechanism similar to that reported with thermal injury. Only nondepolarizing muscle relaxants should be used in patients who are at risk for mucositis. Mucositis should be added to the list of conditions in which succinylcholine is contraindicated. PMID- 11414271 TI - Pitfalls in estimating the effect of interventions in the critically ill using observational study designs. PMID- 11414272 TI - Comment on "the significance of supraspinal control of reflex actions". PMID- 11414273 TI - Dendritic morphogenesis: building an arbor. AB - Neurons are polarized cells with an axon and a dendritic arbor extending from the soma. Although the molecular mechanisms underlying axon guidance are rapidly being elucidated, those that regulate the orientation, morphology, and elaboration of dendritic processes are largely unknown. Several recent papers address these issues, and propose a set of molecular strategies that control dendrite development. This review discusses these papers and what they reveal to us about how cell signaling orchestrates neuronal form and connectivity during development. PMID- 11414274 TI - Amygdala-hippocampus dynamic interaction in relation to memory. AB - Typically the term "memory" refers to the ability to consciously remember past experiences or previously learned information. This kind of memory is considered to be dependent upon the hippocampal system. However, our emotional state seems to considerably affect the way in which we retain information and the accuracy with which the retention occurs. The amygdala is the most notably involved brain structure in emotional responses and the formation of emotional memories. In this review we describe a system, composed of the amygdala and the hippocampus, that acts synergistically to form long-term memories of significantly emotional events. These brain structures are activated following an emotional event and cross-talk with each other in the process of consolidation. This dual activation of the amygdala and the hippocampus and the dynamics between them may be what gives emotionally based memories their uniqueness. PMID- 11414275 TI - Synaptic vesicle endocytosis: calcium works overtime in the nerve terminal. AB - The functions of Ca2+ are many and varied within cells, but in the nerve terminals of neurons it has had a very defined role. That is, the influx of extracellular Ca2+ through voltage-dependent Ca2+ channels stimulates neurotransmitter release by exocytosis. For years this was assumed to be the main role for Ca2+ in this specialized subcellular region. However recent studies have shown that Ca2+ also has multiple roles in synaptic-vesicle endocytosis. This review will present evidence for three Ca2+-dependent and -independent steps; a high-affinity Ca2+-dependent triggering step, a Ca2+-independent maintenance phase, and a low-affinity Ca2+-dependent inhibition step. How the control of endocytosis by Ca2+ might impact on different neuronal functions such as synaptic transmission, the nucleation of SV endocytosis, and the repair of damaged membrane is then discussed. PMID- 11414276 TI - Control of Na+ spike backpropagation by intracellular signaling in the pyramidal neuron dendrites. AB - The integrative function of neurons depends on the somato-dendritic distribution and properties of voltage-gated ion channels. Sodium, potassium, calcium, and hyperpolarization-activated cyclic nucleotide-gated K+ (HCN) channels expressed in the dendrites can be modulated by a number of neurotransmitters and second messenger systems. For example, activation of protein kinases leads to an increase in dendritic excitability by removing a slow inactivation of Na+ channels and decreasing the activity of transient K+ channels in the apical dendrites of hippocampal pyramidal neurons. Consequently, action potentials propagating along the dendrites can be modified significantly by a variety of neuromodulatory synaptic inputs. PMID- 11414278 TI - Components of astrocytic intercellular calcium signaling. AB - It has become evident that astrocytes play major roles in central nervous system (CNS) function. Because they are endowed with ion channels, transport pathways, and enzymatic intermediates optimized for ionic uptake, degradation of metabolic products, and inactivation of numerous substances, they are able to sense and correct for changes in neural microenvironment. Besides this housekeeping role, astrocytes modulate neuronal activity either by direct communication through gap junctions or through the release of neurotransmitters and/or nucleotides affecting nearby receptors. One prominent mode by which astrocytes regulate their own activity and influence neuronal behavior is via Ca2+ signals, which may be restricted within one cell or be transmitted throughout the interconnected syncytium through the propagation of intercellular calcium waves. This review aims to outline the most recent advances regarding the active communication of astrocytes that is encoded by intracellular calcium variation. PMID- 11414277 TI - Regulation of feeding-associated peptides and receptors by nicotine. AB - Although numerous epidemiological studies have provided convincing evidence for the inverse association between tobacco smoking and body weight, the molecular mechanisms underlying this relationship are not well-understood. Nicotine, as a potent secretagogue, could be expected to influence the levels and expression of many classes of neurotransmitters, as well as of cell-membrane constituents linked to neurotransmission, including signal transducers and related effectors. A potentially major group of candidate molecules that could be involved in feeding-related actions of nicotine are the numerous neuropeptides and peptide hormones shown in the past two decades to regulate food intake and energy expenditure. These could include neuropeptide Y (NPY), orexins, leptins, and uncoupling proteins (UCPs). Some of these peptides were already shown to respond to nicotine treatment in terms of regulation of levels and of activity at the level of cell-membrane receptors. The primary objective of this review is to summarize our current understanding of the regulatory effects of nicotine on the food intake and energy expenditure as related to the expression levels of leptin, NPY, orexin, uncoupling proteins, and of NPY and orexin receptors. PMID- 11414279 TI - Carboxylation and anaplerosis in neurons and glia. AB - Anaplerosis, or de novo formation of intermediates of the tricarboxylic acid (TCA) cycle, compensates for losses of TCA cycle intermediates, especially alpha ketoglutarate, from brain cells. Loss of alpha-ketoglutarate occurs through release of glutamate and GABA from neurons and through export of glutamine from glia, because these amino acids are alpha-ketoglutarate derivatives. Anaplerosis in the brain may involve four different carboxylating enzymes: malic enzyme, phosphoenopyruvate carboxykinase (PEPCK), propionyl-CoA carboxylase, and pyruvate carboxylase. Anaplerotic carboxylation was for many years thought to occur only in glia through pyruvate carboxylase; therefore, loss of transmitter glutamate and GABA from neurons was thought to be compensated by uptake of glutamine from glia. Recently, however, anaplerotic pyruvate carboxylation was demonstrated in glutamatergic neurons, meaning that these neurons to some extent can maintain transmitter synthesis independently of glutamine. Malic enzyme, which may carboxylate pyruvate, was recently detected in neurons. The available data suggest that neuronal and glial pyruvate carboxylation could operate at as much as 30% and 40-60% of the TCA cycle rate, respectively. Cerebral carboxylation reactions are probably balanced by decarboxylation reactions,, because cerebral CO2 formation equals O2 consumption. The finding of pyruvate carboxylation in neurons entails a major revision of the concept of the glutamine cycle. PMID- 11414280 TI - Neural activity and survival in the developing nervous system. AB - Recent evidence suggests that blockade of normal excitation in the immature nervous system may have profound effects on neuronal survival during the period of natural cell death. Cell loss following depression of electrical activity in the central nervous system (CNS) may explain the neuropsychiatric deficits in humans exposed to alcohol or other CNS depressants during development. Thus, understanding the role of electrical activity in the survival of young neurons is an important goal of modern basic and clinical neuroscience. Here we review the evidence from in vivo and in vitro model systems that electrical activity participates in promoting neuronal survival. We discuss the potential role of moderate elevations of intracellular calcium in promoting survival, and we address the possible ways in which activity and conventional trophic factors may interact. PMID- 11414281 TI - Spinal-Cord plasticity: independent and interactive effects of neuromodulator and activity-dependent plasticity. AB - Plasticity is one of the most extensively studied aspects in neuroscience. Interest in it has primarily been related to its proposed role in learning and memory and its relevance to adaptive changes following injury. Plasticity can be evoked by changes in molecular, cellular, and synaptic properties, either as a result of activity-dependent effects, or by relatively slow-acting neuromodulatory transmitters. In addition, it is increasingly recognized that the plasticity evoked by these individual effects can be altered by previous inputs and is thus itself plastic. Here, I will review studies in the lamprey spinal cord that have examined individual and interactive activity-dependent and neuromodulator-mediated plasticity. The results show that activity-dependent and neuromodulator-mediated plasticity evoke neuron-and synapse-specific effects at different levels in the spinal cord, and that interactions within and between these effects can evoke dynamic changes in cellular, synaptic, and network plasticity. PMID- 11414284 TI - Trisomy 15 as a single autosomal abnormality in a patient with unclassifiable myelodysplastic syndromes. PMID- 11414282 TI - Closing in on the amyloid cascade: recent insights into the cell biology of Alzheimer's disease. AB - Accumulation of the amyloid-beta (A beta) peptide in the central nervous system (CNS) is considered by many to be the crucial pathological insult that ultimately leads to the development of Alzheimer's disease (AD). Regulating the production and/or aggregation of A beta could therefore be of considerable benefit to patients afflicted with AD. It has long been known that A beta is derived from the proteolytic processing of the amyloid precursor protein (APP) by two enzymatic activities, beta-secretase and gamma-secretase. Recent breakthroughs have led to the identification of the aspartyl protease BACE (beta-site APP cleaving enzyme) as beta-secretase and the probable identification of the presenilin proteins as gamma-secretases. This review discusses what is know about BACE and the presenilins, focusing on their capacity as secretases, as well as the options for therapeutic advancement the careful characterization of these proteins will provide. These findings are presented in the context of the "amyloid cascade hypothesis" and its physiological relevance in AD pathogenesis. PMID- 11414283 TI - Small proteins that modulate calmodulin-dependent signal transduction: effects of PEP-19, neuromodulin, and neurogranin on enzyme activation and cellular homeostasis. AB - Neuromodulin (GAP-43), neurogranin (RC3), and PEP-19 are small acid-stable proteins that bind calcium-poor calmodulin through a loosely conserved IQ-motif. Even though these proteins have been known for many years, much about their function in cells is not understood. It has recently become appreciated that calmodulin activity in cells is tightly controlled and that pools of otherwise free calmodulin are sequestered so as to restrict its availability for activating calcium/calmodulin-dependent enzymes. Neuromodulin, neurogranin, and PEP-19 appear to be major participants in this type of regulation. One way in which they do this is by providing localized increases in the concentration of calmodulin in cells so that the maximal level of target activation is increased. Additionally, they can function as calmodulin antagonists by directly inhibiting the association of calcium/calmodulin with enzymes and other proteins. Although neuromodulin, neurogranin, and PEP-19 were early representatives of the small IQ motif-containing protein family, newer examples have come to light that expand the number of cellular systems through which the IQ-peptide/calmodulin interaction could regulate biological processes including gene transcription. It is the purpose of this review to examine the behavior of neuromodulin, neurogranin, and PEP-19 in paradigms that include both in vitro and in situ systems in order to summarize possible biological consequences that are linked to the expression of this type of protein. The use of protein:protein interaction chromatography is also examined in the recovery of a new calmodulin-binding peptide, CAP-19 (ratMBF1). Consistent with earlier predictions, at least one function of small IQ-motif proteins appears to be that they lessen the extent to which calcium-calmodulin-dependent enzymes become or stay activated. It also appears that these polypeptides can function to selectively inhibit activation of intracellular targets by some agonists while simultaneously permitting activation of these same targets by other agonists. Much of the mechanism for how this occurs is unknown, and possible explanations are examined. One of the biological consequences for a cell that expresses a calmodulin-regulatory protein could be an increased resistance to calcium-mediated toxicity. This possibility is examined for cells expressing PEP-19 and both anatomical and cell-biological data is described. The study of IQ-motif-containing small proteins has stimulated considerable thought as to how calcium signaling is refined in neurons. Current evidence suggests that signaling through calmodulin is not a fulminating and homogenous process but a spatially limited and highly regulated one. Data from studies on neuromodulin, neurogranin, and PEP-19 suggest that they play an important role in establishing some of the processes by which this regulation is accomplished. PMID- 11414285 TI - Carcinoma of the gallbladder: a retrospective review of 99 cases. AB - Since the first description of the carcinoma of the gallbladder, about two centuries ago, this disease has evaded all attempts at early detection and a potential cure. There are only a few studies involving the Indian population, which has a high incidence of gallbladder cancer. Indians are ethnically and culturally different from their Western counterparts, for whom the incidence of this disease is comparatively low. The present study was conducted prospectively on 99 consecutive cases of carcinoma of the gallbladder of 736 patients with biliary diseases admitted to one surgical unit at the University Hospital. The staging, histological type, and grade were correlated with the clinical outcome. Abdominal pain (82.8%) and abdominal mass (73.7%) were the main presenting features. The diagnosis was obtained by ultrasonography (USG) in 93 of 99 cases and confirmed by fine-needle aspiration cytology (FNAC) in 70 patients. Ultrasonography, however, was found to be inadequate for accurate staging of the disease. Based on the TNM classification, 3 (3.0%), 12 (12.1%), 14 (14.1%), 12 (12.1%), and 58 (58.6%) patients had stage 0, I, II, III, and IV disease, with a corresponding mean survival of 28.3, 13.8, 7.5, 5.2, and 3.7 months, respectively. Carcinoma of the gallbladder is difficult to diagnose early due to its vague symptomatology. A high index of suspicion and health education seem to be the only answers available for early detection and improvement of survival. PMID- 11414286 TI - Endoscopic treatment of colorectal polyps and early cancer. AB - To analyze the efficacy and outcome of colonoscopic resection for colorectal neoplastic lesions, we retrospectively reviewed 338 colorectal lesions from 232 patients regarding the clinical profiles, colonoscopic findings, histological findings, complications, and outcome. Morphologically, these lesions were classified into three categories: pedunculated (n = 140), sessile (n = 176); and flat (n = 22). Histological findings of lesions included adenoma (n = 248), carcinoma in situ (n = 17), submucosal carcinoma (n = 2), hyperplastic polyp (n = 57), and inflammatory polyp (n = 14). Neoplastic lesions are generally larger than nonneoplastic lesions (chi2 test, P < 0.05). The incidence of carcinoma was 5.6% of 338 resected lesions. The rate of cancer or high-grade dysplasia in flat polyps was greater than in pedunculated and sessile polyps (13.6 vs 4.54 vs 5.71%; P < 0.05). There were no perforations or deaths after colonoscopic treatment, and only mild bleeding occured in two patients. To date, 19 patients with early colorectal cancer were treated successfully by endoscopy with no recurrence or metastasis. To reduce the incidence and mortality of colorectal cancer, colonoscopic resection is a simple and safe procedure for removing neoplastic lesions. Detailed histological examinations are essential to decide the indications of surgery. PMID- 11414287 TI - Transforming growth factor-alpha precursors in human colon carcinoma cells. AB - Among the proteins of the epidermal growth factor family, transforming growth factor-alpha (TGF-alpha) may be an especially reliable indicator of metastasis or prognosis in human colorectal carcinomas. Moreover, anomalous forms of TGF-alpha have been detected in several tissues of cancer origin, suggesting a role of these forms in the development of the disease. This study was designed to identify the presence of TGF-alpha precursors in different colon cancer cell lines by mean of immunocytochemistry and western blotting techniques. Pro-TGF alpha was detected in all cell lines tested. Staining for pro-TGF-alpha was observed in cytoplasm. Monoclonal antibody to TGF-alpha detected two bands of 20 and 21 kDa. Polyclonal antibody to pro-TGF-alpha revealed five bands ranging from 15 to 24 kDa. All these proteins were also detected in nonmalignant cells expressing a transfected rat pro-TGF-alpha gene. In conclusions, transformation in these human colon carcinoma cells is not due to the presence of anomalous forms of TGF-alpha precursors. PMID- 11414288 TI - Solitary cerebellar metastasis as a presentation for colon cancer: a case report and review of the literature. PMID- 11414289 TI - Metastatic gastric maltoma: a rare cause of obscure gastrointestinal bleeding. PMID- 11414290 TI - Intraduodenal and intrajejunal administration of the herbal medicine, dai-kenchu tou, stimulates small intestinal motility via cholinergic receptors in conscious dogs. AB - The aim of the present study was to study the effect and mechanism of action of intraduodenal and intrajejunal dai-kenchu-to, an herbal medicine clinically effective for uncomplicated postoperative adhesive intestinal obstruction, on upper gastrointestinal motility. Five mongrel dogs were equipped with four strain gauge force transducers on the antrum, duodenum, and proximal and distal jejunum to measure contractile activity. Dai-kenchu-to (0.5, 1.5, and 3.0 g) was administered into the duodenal or proximal jejunal lumen. The effect of atropine, hexamethonium, phentolamine, propranolol, and ondansetron on intraduodenal and intrajejunal dai-kenchu-to-induced contractions was studied. Plasma motilin was measured by specific radioimmunoassay. Intraduodenal and intrajejunal dai-kenchu to induced phasic contractions in the duodenum and proximal jejunum, respectively, and those contractions migrated distally. Phasic contractions induced by intraduodenal and intrajejunal dai-kenchu-to were inhibited by atropine and hexamethonium at all sites. Plasma motilin was not affected by dai kenchu-to. Intraduodenal and intrajejunal dai-kenchu-to stimulates upper gastrointestinal motility at and distal to the administration sites through cholinergic receptors. PMID- 11414291 TI - Intestinointestinal inhibitory reflexes: effect of distension on intestinal slow waves. AB - The motor response induced by intraluminal distension of the small intestine has been well investigated. However, little is known of the myoelectrical response to intraluminal distension. The aim of this study was to investigate the effects of oral- and anal-side distension on jejunal slow waves in dogs. The study was performed in 10 healthy female hound dogs implanted with three pairs of electrodes on the jejunum and an intestinal fistula. The first study session was designed to investigate the effects of anal-side distension on jejunal myoelectrical activity in fasting state. The protocol consisted of a 30-min baseline recording, a 30-min recording during anal-side balloon distension located 5 cm distal to the most distal pair of electrodes, and another 30-min recording after distension. The second session was designed to investigate the effect of oral-side distension with the balloon 5 cm proximal to the most proximal pair of electrodes. Jejunal slow waves were severely impaired by both anal- and oral-side distension. The dominant power was significantly reduced from -2.96 +/- 0.90 dB at baseline to -6.00 +/- 0.64 dB during anal-side distension (P < 0.0005) and from -3.90 +/- 0.85 dB at baseline to -7.17 +/- 0.90 dB during oral side distension (P < 0.001). The percentage of normal 17 to 22-cpm slow waves was significantly decreased from 97.39 +/- 0.88% to 83.48 +/- 3.12% during anal-side distension (P < 0.0005) and from 92.49 +/- 2.42% to 68.80 +/- 7.24% during oral side distension (P < 0.002). The percentage of slow wave coupling was decreased from 95.08 +/- 2.27% to 52.48 +/- 7.73% during anal-side distension (P < 0.0005) and from 84.82 +/- 6.75% to 49.21 +/- 8.91% during oral-side distension (P < 0.001). The instability coefficient of the dominant frequency was significantly increased during anal-side distension. In conclusion, intraluminal distension of the jejunum severely impairs jejunal slow waves. The slow waves on both sides of distension become less coupled, less regular, and are of lower amplitude. PMID- 11414292 TI - How long should a long-term esophageal motility study be? AB - It was the aim of this study to analyze whether a shorter measuring period would render the same diagnostic information on esophageal motility as a circadian measuring period in ambulatory esophageal manometry. In an investigation on normal volunteers (n = 10), patients with gastroesophageal reflux disease without esophageal motility disorders (n = 13), and patients with esophageal motility disorders (n = 14), a comparison was performed between a 5-hr and a 24-hr motility study. An analysis was performed on inter- and intraindividual reproducibility of time periods, prandial phases, and motility sequences (Wilcoxon and Spearman test). There was no significant difference between the two analyzed measuring periods in all three groups with regard to the diagnostic information on esophageal motility in 44 of 45 comparisons for intraindividual variability. A measuring period restricted to 5 hr offers the same diagnostic information on esophageal peristaltic activity as a 24-hr motility study. PMID- 11414293 TI - Role of substance P and calcitonin gene-related peptide in acid-induced augmentation of opossum esophageal blood flow. AB - Increased esophageal blood flow may protect against damaging refluxed gastric juices. We have shown that mast cells, histamine, and nitric oxide increase esophageal blood flow in the opossum during acid perfusion. This study examined the roles of substance P and calcitonin gene-related peptide on acid-induced hyperemia and whether the effects of substance P are mediated by mast cells. The opossum distal esophagus was perfused with saline, acid, or capsaicin while blood flow and histamine release were determined. Neuropeptides and neurokinin antagonists were administered parenterally. Only acid or calcitonin gene-related peptide (not substance P or capsaicin) significantly increased blood flow, which was prevented by neurokinin or calcitonin-gene-related peptide antagonists. Acid, substance P, and capsaicin all increased histamine release. Pretreatment with neurokinin antagonists did not affect acid-induced histamine release. We conclude that calcitonin-gene-related peptide is an important mediator of acid-induced esophageal hyperemia, while substance P plays an indirect role. PMID- 11414294 TI - Esophageal motility of patients with Chagas' disease and idiopathic achalasia. AB - Chagas' disease and idiopathic achalasia have the same neuropathic lesion--the loss of ganglion cells within the esophageal myenteric plexus--with similar clinical, radiologic, and manometric features. However, it is suggested that there are some differences between them. We studied the esophageal motility of 45 patients with Chagas' disease (seven with esophageal dilation), 27 patients with idiopathic achalasia (13 with esophageal dilation), and 40 asymptomatic volunteers. We used the manometric method with continuous perfusion. The lower esophageal sphincter (LES) pressure was measured by the rapid pull-through method. Esophageal contractions was evaluated at 5, 10, and 15 cm above the LES, after 10 swallows of a 5-ml bolus of water alternated with 10 dry swallows. LES pressure was higher in achalasia than in Chagas' disease patients and controls (P < 0.05). Amplitude of contraction was lower in all patient groups compared with controls (P < 0.01) and lower in patients with dilation compared with patients without dilation (P < 0.05). The contraction duration was longer in patients with achalasia than in patients with Chagas' disease and controls (P < 0.05). The percentage of failed contractions was higher in Chagas' disease than in achalasia and controls (P < 0.05), and the percentage of simultaneous contractions was higher in patients with idiopathic achalasia than in patients with Chagas' disease and controls (P < 0.05). The results suggest the possibility that the extent of impairment of esophageal innervation differs between Chagas' disease and idiopathic achalasia. PMID- 11414295 TI - Barriers to gastroesophageal reflux in rats. PMID- 11414296 TI - Role of endogenous nitric oxide and prostaglandin in duodenal bicarbonate response induced by mucosal acidification in rats. AB - Duodenal HCO3- secretion increases in response to mucosal acidification by luminal acid. Although this process is known to be mediated by endogenous prostaglandins (PGs), the role of nitric oxide (NO) in this response has been little studied. We examined the effects of indomethacin and N(G)-nitro-L-arginine methyl ester (L-NAME) on the acid-induced HCO3- secretion in the rat duodenum, together with those on PGE2 generation as well as luminal release of NO metabolites (NOx). A proximal duodenal loop was perfused with saline, and the HCO3- secretion was measured at pH 7.0 using a pH-stat method and by adding 10 mM HCl. Mucosal acidification was performed by exposing the loop to 10 or 100 mM HCl for 10 min. Acidification of the duodenal mucosa stimulated the HCO3- secretion, with concomitant increase of mucosal PGE2 contents and luminal release of NOx, the response being much greater in case of 100 mM HCl. Indomethacin significantly inhibited the acid-induced HCO3- secretion as well as the PGE2 biosynthetic response, without influence on the NOx release. Pretreatment of the animals with L-NAME attenuated both the increase of mucosal PGE2 contents and luminal release of NOx following the acidification, resulting in a marked inhibition of the acid induced HCO3- response, and these effects were significantly antagonized by coadministration of L-arginine. Duodenal HCO3- secretion was also increased by mucosal exposure to NOR-3 (a NO donor), with concomitant increase of PGE2 generation, but these effects were mitigated in the presence of indomethacin. In addition, the duodenal damage caused by mucosal perfusion with 100 mM HCl for 4 hr was markedly aggravated by pretreatment with L-NAME as well as indomethacin. These results suggest that both endogenous NO and PGs are involved in the mechanism for the acid-induced duodenal HCO3- secretion, and that NO may increase the HCO3- secretion by stimulating PG generation. PMID- 11414297 TI - Stimulation of intestinal mucosal afferent nerves increases superior mesenteric artery and decreases mesenteric adipose tissue blood flow. AB - We tested the hypothesis that stimulation of intestinal mucosal afferent nerves produces an increase in superior mesenteric artery (SMA) but a decrease in mesenteric adipose tissue (MAT) blood flow. In anesthetized rats, blood flow in the SMA (pulsed Doppler flowmetry) and MAT (hydrogen gas clearance) was measured simultaneously before and after administration of 0.9% saline, 640 microM capsaicin, or 5% dextrose into the intestinal lumen. The changes in the SMA were 3.8 +/- 3.0, 15.9 +/- 4.0, and 18.8 +/- 7.6%; and those in the MAT, 4.7 +/- 4.0, 11.5 +/- 3.4, and -0.07 +/- 3.4% of baseline, respectively. The data indicate that exposure of the intestinal lumen to an afferent nerve stimulant or nutrient induced a dichotomous pattern of blood flow changes, an increase in the SMA and a reduction in MAT. The capsaicin-sensitive afferent nerves may be instrumental in mediating these energy responses. PMID- 11414298 TI - Effect of bile salts on colonic mucus secretion in isolated vascularly perfused rat colon. AB - Mainly composed of mucins, mucus secreted by goblet cells in the intestinal epithelium is critically involved in the protection of the gastrointestinal mucosa. The hypothesis that bile and some bile salts can induce mucus secretion was tested in the isolated perfused rat colon. Mucus release was evaluated using enzyme-linked immunosorbent assays and supported by histological analysis. Luminal administration of bile extract (1%) provoked mucus secretion in the rat colon. Deoxycholate (0.5-10 mM) induced a dose-dependent increase in rat colonic mucus release. Chenodeoxycholate (10 mM) and hyodeoxycholate (10 mM) also evoked mucus discharge, whereas 10 mM cholate, 10 mM ursodeoxycholate, or Tween-20 did not release mucus. Taurine-conjugated bile salts (deoxycholate, hyodeoxycholate, and chenodeoxycholate) were less potent mucus secretagogues than the corresponding unconjugated forms. The deoxycholate-induced mucus discharge was not altered by pharmacological blockers (tetrodotoxin, atropine), indomethacin, mast cell stabilizers (ketotifen, doxantrazole), H1 histamine receptor antagonist (pyrilamine), or 5-HT receptor antagonists (ketanserin, ondansetron, SDZ 205 557). Our findings suggest that some bile salts, especially in the unconjugated form, may provoke colonic mucus secretion, probably through a direct action on mucus-secreting cells. PMID- 11414299 TI - Pancreaticoduodenal artery aneurysm associated with celiac axis stenosis: combined angiographic and surgical treatment. PMID- 11414300 TI - Changes in cell proliferation and differentiation of adult rat small intestine epithelium after adrenalectomy: kinetic, biochemical, and morphological studies. AB - The effects of 10-day bilateral adrenalectomy on morphometry, proliferation, and apoptosis were determined in the small intestine of 3-month-old Sprague-Dawley rats. The activities of sucrase, lactase, and its respective mRNA, aminopeptidase N, and intestinal alkaline phosphatase were also evaluated. Adrenalectomy lead to partial atrophy and disorganization of the epithelium, with an increased number of goblet and Paneth cells and a reduction of crypt cell proliferation paralleled by a marked increase in villus apoptosis. Biochemical assays revealed that aminopeptidase N and intestinal alkaline phosphatase activities were significantly decreased, whereas disaccharidases were increased by adrenalectomy. The corresponding induction of lactase mRNA suggests an active response of the epithelium. In conclusion, adrenalectomy modified maturation and the differentiation processes of the small intestinal mucosa, especially in the proximal part of the small intestine. This result points to an important role of adrenals and glucocorticoids in the trophic status of the adult small intestinal mucosa. PMID- 11414301 TI - Changes in intraluminal pressure in rat large intestines with aging and effects of dietary fiber. AB - Changes in intraluminal pressure in rat colon with aging and with the effects of dietary fiber were measured. A pressure sensor was inserted into the rat large intestine under endoscopic guidance. The intraluminal pressure curve in the colon was recorded, and the motility index was calculated by this curve. The rats were divided into three groups with a fiber-free diet, a cellulose diet (10% w/w), or a pectin diet (10% w/w). Intraluminal pressure was measured in the proximal, middle, and distal colon at 2, 4, 8, 12, and 16 months after birth. Intraluminal pressure in three sites increased with age and decreased in the latter half of the study. The motility index was lower during the course in the fiber groups, especially the pectin group more than the nonfiber group. This result suggests that long-term ingestion of dietary fiber might have a prophylactic effect on the development of diverticula. PMID- 11414302 TI - Proglucagon-derived peptides in intestinal epithelial proliferation: glucagon like peptide-2 is a major mediator of intestinal epithelial proliferation in rats. AB - Proglucagon-derived peptides have been implicated in the control of intestinal mucosal cell division. To investigate the actions of these peptides on intestinal cell proliferation, different doses of enteroglucagon, oxyntomodulin, glucagon like peptide-1 (GLP-1) and glucagon-like peptide-2 (GLP-2) were tested in male Wistar rats maintained on total parenteral nutrition. Crypt cell proliferation was assessed by the analysis of arrested metaphases in microdissected crypts. Enteroglucagon and oxyntomodulin had no effect on intestinal weight or cell proliferation. GLP-1 had a slight effect on stomach and small intestinal weights and on epithelial cell proliferation in the small and large intestines. GLP-2 infusion dose-dependently increased the weights of the stomach, small intestine, colon, and cecum and increased crypt cell proliferation in the small and large intestines of parenterally fed rats. In orally fed animals, GLP-2 increased intestinal weight but had little effect on proliferation. Therefore, of the proglucagon-derived peptides, GLP-2 appears to be a major mediator of intestinal epithelial proliferation. PMID- 11414303 TI - Recurrent episodes of necrotizing enterocolitis complicating congenital microvillous atrophy. PMID- 11414304 TI - Effectiveness of acupuncture for treatment of childhood constipation. AB - The aim of the present study was to examine the influence of acupuncture on chronic constipation in children and to investigate their basal plasma panopioid level and the changes of this treatment. Seventeen children constipated for at least six months were treated by five weekly placebo acupuncture sessions, followed by 10 weekly true acupuncture sessions. Their parents filled a bowel habit questionnaire. Panopioid activity was measured at time 0 and after 5, 10, and 15 acupuncture sessions. The frequency of bowel movements in males increased more gradually compared to females and reached a maximal improvement only after 10 true acupuncture sessions, from 1.4 +/- 0.6/week to 4.4 +/- 0.6/week and females from 1.4 +/- 0.3/week up to 5.6 +/- 1.2/week. The basal panopioid activity was lower in constipated children as compared to the control population and increased gradually up to control level after 10 true acupuncture sessions. This study is the first to describe a successful treatment by acupuncture of constipated children. PMID- 11414305 TI - Autonomic nervous system function in women with irritable bowel syndrome. AB - Autonomic nervous system (ANS) balance was assessed in women with and without irritable bowel syndrome (IBS) using laboratory tests of function (ie, expiratory/inspiratory ratio, Valsalva, posture changes, and cold pressor) and spectral and nonspectral measures of heart rate variability (HRV). Women with (N = 103) and without IBS (N = 49) were recruited, interviewed, then completed a laboratory assessment and wore a 24-hr Holter monitor Analysis using the entire sample showed little difference between IBS and control women and between subgroups with IBS on either laboratory measures or 24-hr HRV measures. However, analysis restricted to those women with severe IBS symptoms showed quite pronounced differences between two IBS subgroups on 24-hr HRV measures. Parasympathetic tone was significantly lower and ANS balance was significantly higher in the constipation-predominant compared to the diarrhea-predominant group. Subgroups of women with IBS do differ in ANS function as measured by 24-hr HRV; however, these differences are only apparent among women with severe symptoms. These findings point out the importance of considering symptom severity when interpreting studies of IBS. PMID- 11414306 TI - Dose-dependent conjugation of sulfobromophthalein and hepatic transit time in bile fistula rats: role of the microtubule-dependent vesicle pathway. AB - Sulfobromophthalein (BSP) is selectively taken up by the liver and secreted into the bile as unconjugated and conjugated forms. Our previous study demonstrated that unconjugated BSP, but not conjugated BSP, caused the dissociation of biliary lipid secretion from that of bile acids, suggesting that the hepatic BSP conjugation rate partly regulated biliary lipid secretion. To evaluate the mechanisms through which biliary lipid secretion is regulated by exogenous organic anions, we intravenously administered BSP to male Sprague-Dawley rats at various doses either continuously or as a bolus. Then the relationship of the dose of BSP to its conjugation rate, hepatic transit time, and biliary lipid secretion was determined. BSP decreased biliary secretion of cholesterol and phospholipids in a dose-dependent manner without affecting bile acid secretion. In contrast, the proportion of conjugated BSP in bile was associated with the dose. Although the serum clearance of BSP after bolus infusion was constant regardless of the dose administered (50 or 200 nmol/100 g), BSP secretion was delayed with increasing doses: unconjugated BSP was secreted predominantly in the early phase (0-15 min after bolus injection), and conjugated BSP was the predominant form in the late phase (15-30 min). Pretreatment with colchicine reduced the conjugation rate and hepatic transit time of BSP, suggesting that the microtubule-dependent vesicle pathway plays a role in biliary excretion and conjugation of BSP. We conclude that biliary lipid secretion is influenced by organic anions with an affinity for bile acids such as BSP and that this effect is dependent upon the hepatic metabolic rate, i.e., conjugation rate. The hepatic transit time also plays a key role in this process by influencing metabolism. PMID- 11414307 TI - Modifying hepatic phospholipid synthesis associates with biliary phospholipid secretion rate in a transporter-independent manner in rats: relation to canalicular membrane fluidity. AB - Biliary phospholipid secretion is mediated by a multidrug resistance gene product, and its molecular subselection occurs at the site of secretion to modulates bile metastability. The aim of this study was to determine the effect of modifying hepatic phospholipid synthesis on canalicular phospholipid transporter expression and membrane fluidity. Bile-duct cannulation was performed in male Sprague-Dawley rats pretreated with or without intravenous infusion of dimethylethanolamine, an intermediate phospholipid metabolite along the pathway of phosphatidylcholine synthesis of phosphatidylethanolamine N-methylation (0.01 mg/min/100 g body wt) for 15 hr, followed by sodium taurocholate infusion (50 nmol/min/100 g body wt) with or without sulfobromophthalein (50 nmol/min/100 g body wt). Dimethylethanolamine enhanced biliary phospholipid secretion in association with a decrease in biliary phospholipid hydrophobicity. Dimethylethanolamine also increased canalicular membrane fluidity defined by 1,6 diphenyl-1,3,5-hexatriene fluorescence depolarization, whereas the expression of multidrug resistance gene product and multidrug resistance associated protein was unchanged. In contrast, a disproportionate reduction of biliary phospholipid secretion caused by sulfobromophthalein (uncoupling) was enhanced by under the treatment with dimethylethanolamine. In conclusion, the increase in biliary phospholipid secretion and canalicular membrane fluidity without a drastic change of its canalicular transporter by dimethylethanolamine suggests that such a canalicular membrane fluidity facilitates the transporter activity and/or phospholipid molecular movement from the canalicular outer membrane into the bile. A more drastic reduction in phospholipid secretion under sulfobromophthalein-caused uncoupling indicates the possibility of a preferential distribution of relatively hydrophilic phosphatidylcholine molecules to bile salt micelles since sulfobromophthalein is known to reduce the micellar capacity to extract membrane lipids for biliary secretion. PMID- 11414308 TI - Role of PAF in acute liver injury after extended hepatectomy: overexpression of PAF receptor mRNA in Kupffer cells. AB - To clarify the pathophysiology of acute liver injury after extended hepatectomy, we investigated the relationship between the site and levels of expression of platelet activating factor (PAF) receptor mRNA, and the blood levels of endotoxin, cytokines, and prostaglandin E2 (PGE2), using a rat model of 70% hepatectomy. After hepatectomy, a significant rise in serum tumor necrosis factor alpha (TNF-alpha) was observed at 1 hr, the interleukin-6 (IL-6) levels had increased at 3 hr, and PGE2 levels had increased at 6 hr, but there were no significant increases in endotoxin levels in portal vein blood. Overexpression of PAF receptor mRNA in liver tissue was observed as early as 1 hr after hepatectomy, with peak expression occurring at 6 hr, and PAF receptor mRNA was significantly overexpressed in Kupffer cells. The PAF produced in the vicinity of Kupffer cells appears to play a very important role as one of the initial mediators of the acute liver injury that occurs after major hepatectomy. PMID- 11414309 TI - Gender affects reperfusion injury in rat liver. AB - Sex mismatch is a well-known risk factor for chronic rejection of liver allografts, but the mechanisms involved remain unclear. Since experimental observations suggest that female liver is more sensitive to reperfusion injury than male liver, we assessed the influence of gender on oxidative stress. Livers from male and female rats were exposed to warm ischemia and reperfused by an oxygenated buffer. Chemiluminescence was continuously recorded. Reduced and oxidized glutathione and malondialdehyde lactic dehydrogenase values were also determined. Chemiluminescence increased during reperfusion in both groups, but was significantly greater in livers from female rats. Malondyaldehyde and lactic dehydrogenase progressively increased in all animals, reaching significantly greater values in female rats. Livers from female rats showed an increase in all the parameters of oxidative stress compared to male animals. A greater susceptibility to reperfusion injury may be evoked as an alternative mechanism to explain the poor outcome of female organ after liver transplantation. PMID- 11414310 TI - Disturbed synthesis of insulinlike growth factor I and its binding proteins may influence renal function changes in liver cirrhosis. AB - Insulinlike growth factor-1 (IGF-1) is an anabolic hormone synthesized by the liver upon stimulation by growth hormone (GH). IGF-1 exerts important effects on renal hemodynamics and renal sodium handling. The bioactivity of this hormone is influenced by its binding proteins (BP) of which IGF-BP3 favors retention in the capillary lumen while IGF-BP1 facilitates the transport to the target tissues. IGF-BP1 modulates the actions of IGF-1 on target cells including renal tubules. Although a number of reports have dealt with disturbances of the IGF-1/IGF-BP system in cirrhosis, no studies have yet addressed the relationship between alterations in this system and renal function changes in cirrhosis. In the present study we have included 20 patients with cirrhosis and 10 healthy subjects (control group). As compared with the controls, patients showed lower circulating levels of IGF-1 and IGF-BP3, higher IGF-BP1 levels, and a tendency to higher insulinemia and GH values. The index IGF-1 x IGF-BP1/IGF-BP3 (IGF-1-IGF-BP index, reflecting the accessibility of circulating IGF-1 to target cells) was higher in patients with ascites. IGF-1 directly correlated with renal blood flow (P < 0.05), with IGF-BP3 (P < 0.001) and inversely with the Pugh's score (P < 0.02). A negative correlation was found between IGF-1-IGF-BP index and fractional sodium excretion (P < 0.01) and between IGF-BP1 and urinary sodium excretion (P < 0.02). Our findings support the hypothesis that the disturbance of the IGF-1/IGF-BP axis may be related to the degree of renal vasodilation and renal sodium retention in cirrhotic patients. PMID- 11414311 TI - Cyclosporin A is a promising alternative to corticosteroids in autoimmune hepatitis. AB - Autoimmune hepatitis (AIH), a chronic T-cell-mediated liver injury, is treated with corticosteroids with or without Azathioprine. Corticosteroids are not universally effective and have serious side effects. Cyclosporin A was effective in refractory cases. To assess efficacy and safety of Cyclosporin A (Neoral) in induction of remission in AIH patients this study was performed. Nineteen consenting AIH patients (nine treatment-naive) were treated with cyclosporin A in an open label trial and followed for 26 weeks. Liver biopsy was done and hepatitis activity index (HAI) determined at the beginning and end of treatment. Four patients did not complete the study for various reasons. Mean AST and ALT levels decreased from 948.7 +/- 103.5 and 454.8 +/- 354 to 100.6 +/- 111.8 and 78.5 +/- 40.3 (P < 0.03, P < 0.001) respectively. HAI decreased from 15.2 +/- 3.16 to 7.14 +/- 4.01 (P < 0.005). Serum creatinine did not change significantly. In conclusion, low-dose cyclosporin A appears to be safe and effective even in treatment-naive autoimmune hepatitis patients. Randomized controlled trials are warranted. PMID- 11414312 TI - Interdigestive gallbladder emptying, antroduodenal motility, and motilin release patterns are altered in cholesterol gallstone patients. AB - The role of interdigestive gallbladder emptying in gallstone formation is unknown. In fasting healthy subjects, gallbladder emptying is associated with antral phase III of the migrating motor complex (MMC) and high plasma motilin. Therefore, gallbladder volumes and motilin levels were measured during 13 MMC cycles in 10 cholesterol gallstone patients and compared with 20 MMC cycles in 10 healthy subjects. MMC cycle length was longer in gallstone patients than in healthy subjects (158.2 +/- 17.0 vs 105.5 +/- 10.4 min, respectively; P < 0.05), due to longer phase I (39.8 +/- 5.7 vs 17.2 +/- 3.7 min, respectively; P < 0.05). In contrast to healthy subjects, gallstone patients had no significant fluctuations of gallbladder volume during the MMC cycle, and motilin concentrations were not different in MMC cycles with phase III originating in antrum or duodenum. During MMC cycles with phase III originating in the duodenum, motilin levels were twice as high in gallstone patients as in healthy subjects (P < 0.002). In conclusion, cholesterol gallstone patients have an abnormal MMC and motilin release pattern. Their interdigestive gallbladder emptying is reduced and dissociated from the MMC. These disturbances may contribute to gallstone formation. PMID- 11414313 TI - Diagnostic accuracy of fecal elastase 1 assay in patients with pancreatic maldigestion or intestinal malabsorption: a collaborative study of the Italian Society of Pediatric Gastroenterology and Hepatology. AB - Several reports have indicated that fecal elastase-1 (EL-1) determination is a new, sensitive, and specific noninvasive pancreatic function test; however, very few patients with malabsorption due to small intestine diseases have been included in the previous studies. The aim of the study was to compare the diagnostic accuracy of fecal EL-1 and fecal chymotrypsin (FCT) in distinguishing between pancreatic maldigestion and intestinal malabsorption. Three groups of subjects were studied: group A included 49 patients with known cystic fibrosis (25 males, median age 5 years); group B included 43 subjects with various small intestine diseases (17 males, median age 6 years); and group C included 45 children without any history of gastrointestinal disease (22 males, median age 5 years). In all patients, stools were collected for 72 h on a standard diet and fecal EL-1, FCT, and steatocrit tests were performed. Both EL-1 and FCT were below normal limits in all CF patients with pancreatic maldigestion not treated with pancreatic enzyme (100% sensitivity for both assays); El-1, but not FCT, was also below normal in all the CF patients with pancreatic maldigestion treated with pancreatic extracts. Both EL-1 and FCT values in the CF group were significantly lower than in subjects with various small intestinal diseases and in children without any history of gastrointestinal disease (P < 0.0001). FCT, but not EL-1, values showed an inverse statistically significant correlation with steatocrit values in the whole CF group (P < 0.001); FCT was below normal in three of four CF patients with steatorrhea on pancreatic enzyme therapy. Both EL 1 and FCT had 100% specificity when calculated in children without any history of gastrointestinal disease; in contrast, specificity was 86% for EL-1 and 76% for FCT if we considered the control group with small intestinal diseases: low EL-1 was observed in two cases of intestinal giardiasis, two cases of short bowel syndrome, one case of celiac disease, and one case of intestinal pseudobstruction; FCT was abnormal in four cases of intestinal giardiasis, three cases of celiac disease, one case of short bowel syndrome, one case of Crohn's disease, and one case of intestinal pseudobstruction. Diagnostic accuracy was 92% for fecal EL-1 and 82% for FCT. Steatocrit values were over the normal limit in 11 patients with small intestine diseases; in 7/11 of these patients at least one of the pancreatic test results was below the normal limit. In conclusions, in patients with CF, fecal EL-1 determination is not more sensitive than FCT in identifying pancreatic maldigestion; however, fecal EL-1 assay is more specific than FCT determination in distinguishing pancreatic maldigestion from intestinal malabsorption. PMID- 11414314 TI - Differential effects of ET-1, ET-2, and ET-3 on pancreatic microcirculation, tissue integrity, and inflammation. AB - The differential effects of endothelin-1, -2, and -3 (ET-1, ET-2, and ET-3) on pancreatic microcirculation, pancreatic tissue integrity, and an initial inflammatory response, which are three distinct characteristics of acute necrotizing pancreatitis, were investigated in a dose-dependent manner in rats using in vivo microscopy. Red blood cell (RBC) velocity and functional capillary density (FCD) were estimated after topical superfusion of the pancreas with ET-1, ET-2, and ET-3 (100, 10, 1 pmol), revealing that ET-1 (100, 10, 1 pmol) or high ET-2 (100 pmol) and ET-3 (100 pmol) cause a dose-related deterioration of exocrine nutritive pancreatic blood flow. Analysis of pancreatic exocrine tissue damage employing the Spormann score displayed that the ET-mediated microcirculatory impairment was paralleled by dose-dependent tissue damage, which was significant compared to the control group (topical superfusion with 1 ml, saline solution 0.9%). Estimation of pancreatic postcapillary leukocyte accumulation by histomorphologically counting choracetate esterase (CAE) stained leukocytes in 50 high-power fields per animal demonstrated a significant increase in postcapillary accumulation of white blood cells, after topical administration of ET-1, ET-2, and ET-3 compared to controls. In contrast to ET-caused effects on microcirculation and tissue impairment, quantitative analysis of postcapillary leukocyte accumulation revealed the most pronounced effect after ET-2 administration but not after ET-1 administration. This demonstrates that ET-1, ET 2, and ET-3 are all able to mediate microcirculatory impairment, tissue damage, and inflammation. However, ET-3-induced damaging effects are less pronounced, while ET-1 most severely alters microcirculation and ET-2 preferentially induces leukocyte-dependent inflammation. PMID- 11414315 TI - Directed evolution and the creation of enantioselective biocatalysts. AB - Directed evolution has emerged as a key technology to generate enzymes with new or improved properties that are of major importance to the biotechnology industry. A directed evolution approach starts with the identification of a target enzyme to be optimized and the cloning of the corresponding gene. An efficient expression system is needed before the target gene is subjected to random mutagenesis and/or in vitro recombination, thereby creating molecular diversity. Subsequently, improved enzyme variants are identified, preferably after being secreted into culture medium, by screening or selection for the desired property. The genes encoding the improved enzymes are then used to parent the next round of directed evolution. Enantioselectivity is a biocatalyst property of major biotechnological importance that is, however, difficult to deal with. We discuss recent examples of creating enantioselective biocatalysts by directed evolution. PMID- 11414316 TI - Infrared-thermographic screening of the activity and enantioselectivity of enzymes. AB - The infrared radiation caused by the heat of reaction of an enantioselective enzyme-catalyzed transformation can be detected by modern photovoltaic infrared (IR)-thermographic cameras equipped with focal-plane array detectors. Specifically, in the lipase-catalyzed enantioselective acylation of racemic 1 phenylethanol, the (R)- and (S)-substrates are allowed to react separately in the wells of microtiter plates, the (R)-alcohol showing hot spots in the IR thermographic images. Thus, highly enantioselective enzymes can be identified at kinetic resolution. PMID- 11414317 TI - Two-step process for ketocarotenoid production by a green alga, Chlorococcum sp. strain MA-1. AB - The production of ketocarotenoids (KCs) from Chlorococcum sp. strain MA-1 was investigated by a two-step process. In the first step, 18 g biomass l(-1) was achieved by feeding glucose to the heterotrophic cultures; in the second step, the high-density cultures were treated with light illumination or chemical stress in dark, respectively, to induce KC synthesis. Light-treated cultures could produce 103 mg total KCs l(-1) and 32 mg astaxanthin l(-1), three times higher than those from chemical-treated cultures, in the 10 days of induction. The percentages of individual KCs (hydroxyechinenone, canthaxanthin, adonirubin and astaxanthin) in the total KCs were not markedly influenced by the different stress conditions. The developed two-step process provides a feasible strategy for commercial production of ketocarotenoids by the green microalga, Chlorococcum sp. strain MA-1. PMID- 11414318 TI - Biotransformation of limonene by Pseudomonas putida. AB - From a study of three fungal and 15 bacterial strains, it was observed that Pseudomonas putida MTCC 1072 oxidized limonene with the highest efficiency of. Fermentation of limonene by P. putida MTCC 1072 was conducted for 120 h at 30 degrees C at a fixed pH of 5.0. Major bioconversion products were isolated and characterized by Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR) spectroscopy, and by elemental analysis. The bioconversion products were identified as perillyl alcohol and p-menth-1-ene-6,8-diol, and under optimum conditions the yields were 36% and 44%, respectively (a rate kinetic model indicated corresponding limiting yields of 44% and 56%). No further degradation of the products was observed using this bacteria. PMID- 11414319 TI - High cell density cultivation of Rhodococcus opacus for lipid production at a pilot-plant scale. AB - The triacylglycerol (TAG)-accumulating bacterium Rhodococcus opacus strain PD630 was investigated with respect to the fermentative production of TAGs consisting of an unusually high fraction of fatty acids with an odd-number of carbon atoms and unsaturated monoenic fatty acids from sugar beet molasses and sucrose. Fed batch fermentations were optimized at the 30-1 scale in a stirred tank bioreactor at 30 degrees C using a mineral salts medium, which contained sugar beet molasses and sucrose as sole carbon sources. Approximately 37.5 g cell dry matter (CDM) per liter was the highest cell density that was obtained at that scale with a TAG content in the cells of 52%. This fermentative process was also applied to a 500 1 pilot-plant scale. Cell densities as high as 18.4 g CDM per liter were obtained, and 42% of the sucrose present in the medium was converted into cell mass which consisted of 38.4% TAGs. PMID- 11414320 TI - Growth and differentiation of permanent and secondary mouse myogenic cell lines on microcarriers. AB - Myogenesis involves the determination of progenitor cells to myoblasts, their fusion to yield multinuclear myotubes, and the maturation of myotubes to muscle fibres. This development is reflected in a time pattern of gene expression, e.g. of genes coding for desmin, the myogenic factors myogenin and myoD, the acetylcholine receptor alpha-subunit and the muscular chloride channel CIC-1. We attempted to improve yields and myogenic differentiation in culture by using three-dimensional microcarrier systems. Out of a variety of carriers tested in stationary cultures, collagen-coated dextran Cytodex3 beads proved optimal for the proliferation and differentiation of the murine myogenic cell line C2C12. With C2C12 myoblasts in stationary and stirred systems (Spinner- and SuperSpinner flasks), surface adherence, differentiation into myotubes and expression of muscle-specific mRNAs on Cytodex3 beads were the same as in conventional cultures. Other carriers tested (DEAE cellulose, glass, plastic, cellulose, polyester) did not support growth and differentiation of C2C12 cells. The secondary mouse myogenic stem cells M12 and M2.7-MDX proliferated and differentiated well in stationary Cytodex3 cultures, but no differentiation occurred in Spinner flasks. As indicated by light and scanning electron microscopy, C2C12 myotubes formed not only on but also in between Cytodex beads. The secondary cell lines may succumb to shear forces under these conditions. PMID- 11414321 TI - Efficient transformation of filamentous fungus Pleurotus ostreatus using single strand carrier DNA. AB - The effects of carrier DNAs on the transformation of the basidiomycete Pleurotus ostreatus were analyzed. When lambda phage DNA was added to a transformation mixture containing protoplasts and CbxR vector plasmid, an increased number of drug-resistant transformants was observed on a screening plate containing 2 microg carboxin/ml. The highest efficiency (about 200 transformants/microg vector plasmid) was obtained by the addition of heat-denatured lambda DNA, which gave yields approximately 50-fold higher than the control experiment without a carrier DNA. To our knowledge, this is the first report on enhancement in transformation efficiency of fungal protoplasts by single strand carrier DNA. PMID- 11414322 TI - Homologous expression of recombinant manganese peroxidase genes in ligninolytic fungus Pleurotus ostreatus. AB - Pleurotus ostreatus is a white-rot fungus known as an efficient degrader of lignin and also various organo-pollutants. Using a DNA-mediated transformation system, a molecular breeding approach to isolate over-producers of a manganese peroxidase isozyme, MnP3, was carried out. Recombinant mnp3 constructs under the control of P. ostreatus sdi1 expression signals were introduced into the wild type P. ostreatus strain by co-transformation with a carboxin-resistant vector plasmid, pTM1. One of the recombinants obtained by a mating between two monokaryotic transformants, TMG9-C1, showed a several times higher level of MnP activity than the wild-type control in the early stage of liquid culture. Predominant transcription of the recombinant mnp3 in the strain was demonstrated by RT-PCR experiments. This is the first report of a genetically modified P. ostreatus strain with an expression system for recombinant genes. PMID- 11414323 TI - High-rate 3-methylcatechol production in Pseudomonas putida strains by means of a novel expression system. AB - The bioconversion of toluene into 3-methylcatechol was studied as a model system for the production of valuable 3-substituted catechols in general. For this purpose, an improved microbial system for the production of 3-methylcatechol was obtained. Pseudomonas putida strains containing the todC1C2BAD genes involved in the conversion of toluene into 3-methylcatechol were used as hosts for introducing extra copies of these genes by means of a novel integrative expression system. A construct was made containing an expression cassette with the todC1C2BAD genes cloned under the control of the inducible regulatory control region for naphthalene and phenanthrene degradation, nagR. Introducing this construct into wild-type P. putida F1, which degrades toluene via 3 methylcatechol, or into mutant P. putida F107, which accumulates 3 methylcatechol, yielded biocatalysts carrying multiple copies of the expression cassette. As a result, up to 14 mM (1.74 g l(-1)) of 3-methylcatechol was accumulated and the specific production rate reached a level of 105 micromol min( 1) g(-1) cell dry weight, which is four times higher than other catechol production systems. It was shown that these properties were kept stable in the biocatalysts without the need for antibiotics in the production process. This is an important step for obtaining designer biocatalysts. PMID- 11414324 TI - Deletion of a cytotoxic, N-terminal putatitive signal peptide results in a significant increase in production yields in Escherichia coli and improved specific activity of Cel12A from Rhodothermus marinus. AB - The thermostable cellulase Cel12A from Rhodothermus marinus was produced at extremely low levels when expressed in Escherichia coli and was cytotoxic to the cells. In addition, severe aggregation occurred when moderately high concentrations of the enzyme were heat-treated at 65 degrees C, the growth optimum of R. marinus. Sequence analysis revealed that the catalytic module of this enzyme is preceded by a typical linker sequence and a highly hydrophobic putative signal peptide. Two deletion mutants lacking this hydrophobic region were cloned and successfully expressed in E. coli. These results indicated that the N-terminal putative signal peptide was responsible for the toxicity of the full-length enzyme in the host organism. This was further corroborated by cloning and expressing the hydrophobic N-terminal domain in E. coli, which resulted in extensive cell lysis. The deletion mutants, made up of either the catalytic module of Cel12A or the catalytic module and the putative linker sequence, were characterised and their properties compared to those of the full-length enzyme. The specific activity of the mutants was approximately three-fold higher than that of the full-length enzyme. Both mutant proteins were highly thermostable, with half-lives exceeding 2 h at 90 degrees C and unfolding temperatures up to 103 degrees C. PMID- 11414325 TI - Pre-termination in aflR of Aspergillus sojae inhibits aflatoxin biosynthesis. AB - The aflR gene product is the main transcriptional regulator of aflatoxin biosynthesis in Aspergillus parasiticus and Aspergillus flavus. Although A. sojae strains do not produce aflatoxins, they do have an aflR homologue. When compared with the aflR of A. parasiticus, the A. sojae gene contains two mutations: an HAHA motif and a premature stop codon. To investigate the functionality of the A. sojae aflR gene product, we used a GAL4 one-hybrid system in yeast. The transcription-activating activity of AflR from A. sojae was 15% of that from A. parasiticus. The introduction of an additional aflR from A. sojae into an A. parasiticus strain did not affect aflatoxin productivity. A hybrid aflR comprising the amino-terminal region of A. sojae aflR and the carboxy-terminal region of A. parasiticus aflR suppressed the effect associated with pre termination of the A. sojae AflR. We conclude that the premature stop codon of the A. sojae aflR is the key to its functionality and leads to prevention of aflatoxin biosynthesis through loss of the transcription of aflatoxin biosynthesis-related genes. PMID- 11414326 TI - Synthesis of optically pure ethyl (S)-4-chloro-3-hydroxybutanoate by Escherichia coli transformant cells coexpressing the carbonyl reductase and glucose dehydrogenase genes. AB - The asymmetric reduction of ethyl 4-chloro-3-oxobutanoate (COBE) to ethyl (S)-4 chloro-3-hydroxybutanoate ((S)-CHBE) was investigated. Escherichia coli cells expressing both the carbonyl reductase (S1) gene from Candida magnoliae and the glucose dehydrogenase (GDH) gene from Bacillus megaterium were used as the catalyst. In an organic-solvent-water two-phase system, (S)-CHBE formed in the organic phase amounted to 2.58 M (430 g/l), the molar yield being 85%. E. coli transformant cells coproducing S1 and GDH accumulated 1.25 M (208 g/l) (S)-CHBE in an aqueous monophase system by continuously feeding on COBE, which is unstable in an aqueous solution. In this case, the calculated turnover of NADP+ (the oxidized form of nicotinamide adenine dinucleotide phosphate) to CHBE was 21,600 mol/mol. The optical purity of the (S)-CHBE formed was 100% enantiomeric excess in both systems. The aqueous system used for the reduction reaction involving E. coli HB101 cells carrying a plasmid containing the S1 and GDH genes as a catalyst is simple. Furthermore, the system does not require the addition of commercially available GDH or an organic solvent. Therefore this system is highly advantageous for the practical synthesis of optically pure (S)-CHBE. PMID- 11414327 TI - Molecular cloning of a putative divalent-cation transporter gene as a new genetic marker for the identification of Lactobacillus brevis strains capable of growing in beer. AB - Random amplified polymorphic DNA (RAPD) PCR analysis of Lactobacillus brevis isolates from breweries revealed that one of the random primers could distinguish beer-spoilage strains of L. brevis from nonspoilage strains. The 1.1-kb DNA fragment amplified from all beer-spoilers included one open reading frame, termed hitA (hop-inducible cation transporter), which encodes an integral membrane protein with 11 putative trans-membrane domains and a binding protein-dependent transport signature of a non-ATP binding membrane transporter common to several prokaryotic and eukaryotic transporters. The hitA polypeptide is homologous to the natural resistance-associated macrophage protein (Nramp) family characterized as divalent-cation transport proteins in many prokaryotic and eukaryotic organisms. Northern blot analysis indicated that the hitA transcripts are expressed in cells cultivated in MRS broth supplemented with hop bitter compounds, which act as mobile-carrier ionophores, dissipating the trans-membrane pH gradient in bacteria sensitive to the hop bitter compounds by exchanging H+ for cellular divalent cations such as Mn2+. This suggests that the hitA gene products may play an important role in making the bacteria resistant to hop bitter compounds in beer by transporting metal ions such as Mn2+ into cells that no longer maintain the proton gradient. PMID- 11414328 TI - Improved antifungal activity of a mutant of Trichoderma harzianum CECT 2413 which produces more extracellular proteins. AB - Trichoderma harzianum is a well-known biological control agent against fungal plant diseases. In order to select improved biocontrol strains from Trichoderma harzianum CECT 2413, a mutant has been isolated for its ability to produce wider haloes than the wild type, when hydrolysing pustulan, a polymer of beta-1,6 glucan. The mutant possesses between two and four times more chitinase, beta-1,3- and beta-1,6-glucanase activities than the wild type, produces about three times more extracellular proteins and secretes higher amounts of a yellow pigment (alpha-pyrone). This mutant performed better than the wild type during in vitro experiments, overgrowing and sporulating on Rhizoctonia solani earlier, killing this pathogen faster and exerting better protection on grapes against Botrytis cinerea. PMID- 11414329 TI - Insights into the genetic diversity of initial dioxygenases from PAH-degrading bacteria. AB - Alpha subunit genes of initial polyaromatic hydrocarbon (PAH) dioxygenases were used as targets for the PCR detection of PAH-degrading strains of the genera Pseudomonas, Comamonas and Rhodococcus which were obtained from activated sludge or soil samples. Sequence analysis of PCR products from several Pseudomonas strains showed that alpha subunits (nahAc allele) of this genus are highly conserved. PCR primers for the specific detection of alpha subunit genes of initial PAH dioxygenases from Pseudomonas strains were not suitable for detecting the corresponding genes from the genera Comamonas and Rhodococcus. Southern analysis using a heterologous gene probe derived from the P. putida OUS82 PAH dioxygenase alpha subunit identified segments of the PAH-degradation gene cluster from C. testosteroni strain H. Parts of this gene cluster containing three subunits of the initial PAH dioxygenase were isolated. These three subunits [ferredoxin (pahAb), alpha (pahAc) and beta (pahAd) subunit] were amplified by PCR as one fragment and expressed in Escherichia coli DH5alpha, resulting in an active initial dioxygenase with the ability to transform indole and phenanthrene. The DNA sequence alignment of alpha subunits from C. testosteroni H and various PAH-degrading bacteria permitted the design of new primers and oligonucleotide probes which are useful for the detection of the initial PAH dioxygenases from strains of Pseudomonas, Comamonas and Rhodococcus. PMID- 11414330 TI - Biodegradation of synthetic and naturally occuring mixtures of mono-cyclic aromatic compounds present in olive mill wastewaters by two aerobic bacteria. AB - Two bacterial strains, Ralstonia sp. LD35 and Pseudomonas putida DSM 1868, were assayed for their ability to degrade the monocyclic aromatic compounds commonly found in olive mill wastewaters (OMWs). The goal was to study the possibility of employing the two strains in the removal of these recalcitrant and toxic compounds from the effluents of anaerobic treatment plants fed with OMWs. At first, the two strains were separately assayed for their ability to degrade a synthetic mixture of nine aromatic acids present in OMWs, both in growing- and resting-cell conditions. Then, due to the complementary activity exhibited by the two strains, a co-culture of the two bacteria was tested under growing-cell conditions for degradation of the same synthetic mixture. Finally, the degradation activity of the co-culture on two fractions was studied. Both fractions one deriving from natural OMWs through reverse osmosis treatment and containing low-molecular weight organic molecules, and the other obtained from an anaerobic lab-scale treatment plant fed with OMWs, were rich in monocyclic aromatic compounds. The co-culture of the two strains was able to biodegrade seven of the nine components of the tested synthetic mix (2, 6-dihydroxybenzoic acid and 3, 4, 5-trimethoxybenzoic acid were the two undegraded compounds). In addition, an efficient biodegrading activity towards several aromatic molecules present in the two natural fractions was demonstrated. PMID- 11414331 TI - Efficiency of naphthalene and salicylate degradation by a recombinant Pseudomonas putida mutant strain defective in glucose metabolism. AB - Metabolically engineered microorganisms may have tremendous potential in removing toxic compounds from nature. In general, microorganisms prefer to utilize simpler carbon sources over toxic compounds when both are present in an environment and, therefore, the presence of simpler carbon sources may greatly reduce the efficiency of a microorganism towards toxic compounds. If a microorganism is prevented from utilizing simpler carbon sources, thereby making it totally dependent upon the toxic compounds, it should increase the specificity for and efficiency of degradation of the toxic compounds in the presence of other, simpler carbon sources. To test this hypothesis, the efficiency of naphthalene and salicylate degradation in the presence of glucose by a recombinant Pseudomonas putida strain mutated in glucose metabolism was determined and compared to the non-mutated strain. Results obtained indicate that the impairment of glucose metabolism leads to better degradation of naphthalene and salicylate in the presence of glucose. PMID- 11414332 TI - Screening for microorganisms that produce only endo-inulinase. AB - Sixteen fungal strains reported in the literature as endo-inulinase producers and three bacterial strains, isolated from the dahlia rhizosphere, were analysed for endo-inulinase production. From four isolated strains (one fungus and three bacteria) the results were evaluated in terms of substrate consumption, cell growth and production of endo-inulinases. All three bacterial strains were sole endo-inulinase producers and, among these, strain Paenibacillus sp. CDB 003 was the most suitable for endo-inulinase production, as this enzyme produced inulobiose as the principal substrate as well as inulo-oligosaccharides with polymerisation degrees of 3-5. PMID- 11414333 TI - Copper removal from aqueous solution using Aspergillus niger mycelia in free and polyurethane-bound form. AB - This study assesses the ability of mycelia of Aspergillus niger B-77 (both free and immobilized on polyurethane foam) to remove copper from single-ion solution. All experiments were conducted using 0.5 mM solutions of CuSO4.5H2O. Mycelia immobilized on polyurethane foam cells showed a three-fold increase in uptake, compared with that of free cells. The efficiency of copper removal (mg Cu2+ removed/mg Cu2+ added) in a column system reached more than 99% before the break through point was attained. PMID- 11414334 TI - Sugar composition of biofilms produced by paper mill bacteria. AB - Biofilms of paper mill bacteria were cultivated in paper mill white water simulating conditions on glass slides or stainless steel coupons in a laboratory culture system. The sugar content and composition of the biofilms were analysed and compared with the sugar composition of paper mill slimes. Acid methanolysis followed by gas chromatography revealed that Burkholderia was the major biofilm producer in pure culture, producing up to 50 microg of biofilm sugar cm(-2) in 5 days in rich medium and 10 microg in paper mill simulating medium. A mixture of simulated paper mill water with a culture medium yielded more biofilm (100 microg cm(-2)) than either of the media alone, so the biofilm accumulation was not proportional to the available substrate. More biofilm accumulated on stainless steel coupons than on glass slides, and the steel-coupon biofilms contained slightly more uronic acids. The biofilm sugars contained mainly galactose, glucose, mannose, and rhamnose. In paper mill medium, the Burkholderia biofilm contained more galactose and glucose, and less rhamnose, than in rich laboratory medium. The sugar composition of paper mill slimes was quite similar to those of steel-cultured Burkholderia cepacia biofilms. This suggests that Burkholderia cepacia is responsible for much of the slime in the paper mill. PMID- 11414335 TI - Enhancing the start-up of a UASB reactor treating domestic wastewater by adding a water extract of Moringa oleifera seeds. AB - Water extract of Moringa oleifera seeds (WEMOS) was used to enhance the start-up of a self-inoculated upflow anaerobic sludge blanket (UASB) reactor treating raw domestic wastewater. Two reactors labelled control (RC) and WEMOS addition (RM) were started without special inoculum. Both reactors were fed continuously for 22 weeks with domestic wastewater containing an average total chemical oxygen demand (COD) of 320 mg O2/l and suspended solid (SS) of 165 mg/l. The reactors operated during the entire experimental period at 29 degrees C and at a hydraulic retention time (HRT) of 4 h. The RM reactor received 2 ml WEMOS per litre of influent. WEMOS solution was prepared on the basis of 2.5% (w/v) ground M. oleifera seeds in water. The results of 22 weeks' operation showed an improvement in the performance of the RM compared to that of the RC. The dosage of WEMOS in the feed (1) shortened the biological start-up period by 20%, (2) increased acidogenic and methanogenic activity by a factor of 2.4 and 2.2 respectively, (3) increased the specific biogas production by a factor of 1.6, (4) favoured fast growth of the sludge bed, and (5) allowed the aggregation of coccoid bacteria and growth of microbial nuclei, which are precursors of anaerobic granulation. PMID- 11414336 TI - Selection of screening items for alcohol abuse and alcohol dependence among Mexicans and Mexican Americans in the emergency department. AB - OBJECTIVE: This article reports on the selection of screening items to detect Mexican or Mexican-American patients in the emergency department (ED) who have alcohol problems and could benefit from an intervention or a referral for treatment. Items are tested against the Rapid Alcohol Problems Screen (RAPS), which has been optimized from standard screening instruments and has outperformed these instruments. METHOD: The performance of individual items from standard screening instruments (CAGE, TWEAK, AUDIT, TRAUMA and BMAST) against International Classification of Diseases, Tenth Revision and Diagnostic and Statistical Manual, Fourth Revision criteria for alcohol abuse and dependence was evaluated in a merged probability sample (N = 869; 72% men) of 537 ED patients from three hospitals in Pachuca, Mexico, and 332 Mexican-American ED patients in Santa Clara County, CA. Logistic regression and tree-classification models were used for item selection. RESULTS: We found a prevalence of 15% for alcohol dependence and a prevalence of 28% for alcohol abuse or dependence in the merged sample. The RAPS items did not perform as well in terms of sensitivity (93%) as the optimal five items identified in these analyses (sensitivity = 98%) for alcohol dependence, but did demonstrate better specificity (79%) than the optimal five items (65%), which is an important consideration in a time of cost containment. Both sets of items showed better sensitivity and positive predictive value but similar Receiver Operating Characteristic values for respondents in the high acculturation group compared to those at other levels of acculturation. Differences in positive predictive value across all subgroups tended to increase at increased cutpoints, especially for the RAPS. CONCLUSIONS: These analyses suggest that the RAPS performs favorably compared to those items optimized, in this sample of Hispanic ED patients. Based on comparative item performance in these analyses, the RAPS may hold promise as a useful tool for screening for alcohol dependence, but requires further evaluation as a stand-alone instrument in comparison with other standard screening instruments. PMID- 11414337 TI - Gender and reported health problems in treated alcohol dependent Alaska natives. AB - OBJECTIVE: An ongoing study of phenotypes of alcohol dependence among Alaska Natives provides the opportunity to investigate gender differences in reported health-related problems among alcohol dependent clients in three residential programs in Anchorage, Alaska. METHOD: Clinical assessment information was obtained on 469 (263 male) subjects from consecutive admissions to each of three treatment programs. The average (SD) age of the sample was 33.7 (8.4) years. Patterns of substance use, comorbid psychopathology, overall health status, alcohol and other drug withdrawal symptoms, and psychological and physical consequences of alcohol and other drug use were examined. RESULTS: Male and female subjects reported similar experiences with alcohol-related health problems, including symptoms of withdrawal and the psychological and physical consequences of chronic alcohol abuse. However, women were significantly more likely to have lifetime diagnoses of major depression and cocaine dependence, whereas men were more likely to have lifetime diagnoses of antisocial personality disorder and marijuana dependence. Women reported a lower overall health status, more medication use and pain complaints, and more negative consequences of cocaine abuse and withdrawal than did men. CONCLUSIONS: Both men and women within this sample of inpatient alcohol-dependent Alaska Natives were found to have a similar early onset and rapid progression to DSM-III-R alcohol dependence, and to report a similar prevalence of alcohol-related psychological and physical problems. Reports by women of more pain symptoms, more medication use and more negative health consequences related to their cocaine abuse, compared with men in this alcohol dependent sample, suggests additional considerations for treatment planning and intervention. PMID- 11414338 TI - Validity of the CAGE questionnaire in an American Indian population. AB - OBJECTIVE: This study evaluated the performance of the CAGE questionnaire (a set of four questions about alcoholism) in an American Indian population. METHOD: We analyzed data from a cross-sectional study of 275 individuals (179 women) aged 21 years or older. Alcohol dependence was diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders, Third Edition-Revised (DSM-III-R), based on a detailed psychiatric interview using the Schedule for Affective Disorders and Schizophrenia-Lifetime Version. Accuracy of the CAGE questionnaire was quantified as sensitivity, specificity, likelihood ratios and the area under receiver operating characteristics (ROC) curves, using the DSM-III-R diagnosis as the reference. RESULTS: Of participants interviewed, 85% of men and 53% of women had a diagnosis of alcohol dependence by DSM-III-R. A CAGE score of > or = 2 had a sensitivity and specificity of 68% and 93%, respectively, in men and 62% and 79% in women, for the diagnosis of alcohol dependence. CAGE scores of 0, 1 and > or = 2 were associated with likelihood ratios of 0.3, 0.3 and 9.5, respectively, in men and 0.4, 0.7 and 1.5 in women. The area under the ROC curve was 81% for men and 75% for women. CONCLUSIONS: These findings suggest that the CAGE questionnaire is a valid screening method, in this population, for identifying people likely to have alcohol dependence. PMID- 11414339 TI - Differences in drinking patterns among Ashkenazic and Sephardic Israeli adults. AB - OBJECTIVE: Since its foundation in 1948, Israel has received large waves of immigrants, mainly from Europe (Ashkenazic Jews, or Ashkenazim) and from North Africa and other Middle Eastern countries (Sephardic Jews. or Sephardim). In Israeli society, Ashkenazic Jews are an advantaged ethnic group, whereas Sephardic Jews are relatively disadvantaged. Little is known about the differences in drinking patterns between these two groups. The relationship between ethnicity and alcohol consumption is investigated in a 1995 data set from a sample of 4,984 subjects (60% women). METHOD: The data were collected as part of a national Israeli survey. Standardized questions covered drinking and becoming drunk in the last 12 months and drinking in the last 30 days. Unadjusted odds ratios (ORs) indicated the association of group status with the drinking variables. ORs adjusted for potential confounders were created with logistic regression. RESULTS: Unadjusted ORs indicated a negative association between all alcohol measures and Sephardic group status. ORs adjusted for such factors as socioeconomic status and religiosity produced similar results. CONCLUSIONS: This study indicates that Sephardim were less likely to drink or become drunk than were Ashkenazim. Further work is required to determine if these differences are stable or changing over time and whether such differences can be attributed to cultural or genetic factors. Similarities to U.S. patterns are discussed. PMID- 11414340 TI - Alcohol use among college students: the effects of prior problem behaviors and change of residence. AB - OBJECTIVE: This article examines the relationship between prior problem behaviors and change in residence on alcohol use patterns among college students. METHOD: Measures of alcohol consumption (e.g., drinking frequency, average consumption and frequency of heavy episodic drinking were related to residence patterns and prior problem behaviors (e.g., conduct problems. illicit substance involvement and early age at onset of alcohol consumption). Subjects (N = 2,465; 51% women) were a subsample drawn from the National Longitudinal Study of Labor Market Experience in Youth. The analysis was conducted using a linear growth model for continuous outcomes with time-invariant and time-varying covariates for each of the drinking measures. RESULTS: The results of the structural equation analysis yielded significant and direct effects related to residence patterns and prior problem behaviors. Problem behaviors were related to drinking measures; however, there was no evidence for a mediational hypothesis. Neither was there systematic evidence that the relationship between prior problem behavior and alcohol use was mediated by residence patterns. The analysis of change in residence was related to both time-specific and longer-term influences on alcohol use. CONCLUSIONS: The presence of direct and independent effects for both dispositional and high-risk environmental factors in collegiate drinking practices support targeted and diverse strategies for prevention activities. PMID- 11414341 TI - Disinhibition and expectancy in risk for alcohol use: comparing black and white college samples. AB - OBJECTIVE: This study tested several predictions of the "acquired preparedness" model in both black and white samples of college students. The acquired preparedness model holds that trait disinhibition affects alcohol-related learning and, ultimately, alcohol use. This model maintains that the reward focus typical of disinhibited individuals increases the likelihood of forming overly positive expectancies about the effects of alcohol. Alcohol expectancy, then, acts as a mediator of the relationship of disinhibition and drinking behavior. METHOD: Participants (N = 479, 341 women) were 279 white and 200 black college students. Self-reported alcohol expectancy, disinhibition and drinking behavior were assessed. Covariance structure analysis was used to test hypotheses separately for each sample, controlling for socioeconomic status. RESULTS: Black participants scored significantly lower on disinhibition, expectancy and drinking. However, invariance testing indicated that the relationships between these variables were not different across groups. Results were consistent with the stated hypotheses in both samples--alcohol expectancy functioned as a mediator of the disinhibition-drinking relationship. Results did not differ across expectancy content. CONCLUSIONS: These results provide support for the validity of the acquired preparedness model. Despite mean differences in risk and drinking levels between black and white samples, psychosocial learning appears to mediate the influence of disinhibition on drinking for both groups. PMID- 11414342 TI - Successful remission of late-life drinking problems: a 10-year follow-up. AB - OBJECTIVE: This study sought to determine (1) the rate and predictors of long term remission among a sample of untreated late-life problem drinkers and (2) whether successfully remitted older problem drinkers attain levels of functioning and life contexts comparable to those of lifetime nonproblem drinkers at a 10 year follow-up. METHOD: We compared 140 older baseline problem drinkers who were successful in achieving long-term remission to 184 baseline problem drinkers whose drinking problems did not remit over the course of 10 years and to 339 lifetime nonproblem drinkers, on functioning and life contexts at baseline and at 4- and 10-year follow-ups. RESULTS: Being female, having more recent onset of drinking problems, fewer and less severe drinking problems, friends who approved less of drinking, and drinking less and drinking less frequently at baseline predicted long-term remission. In many regards, long-term remitted problem drinkers attained levels of functioning and life context similar to those of lifetime nonproblem drinkers. However, remitted problem drinkers continued to report more incipient drinking problems, depressive symptoms, health and financial stressors, psychoactive medication use, reliance on avoidance coping strategies and less social support from friends than did lifetime nonproblem drinkers at the 10-year follow-up. CONCLUSIONS: About a third (30%) of an untreated sample of late-life problem drinkers succeeded in attaining stable, long-term remission. The functioning and life contexts of untreated remitted problem drinkers improved significantly over time; however, some deficits persisted at follow-up. PMID- 11414343 TI - Effects of age and alcoholism on sleep: a controlled study. AB - OBJECTIVE: Sleep problems are common, costly and potentially fatal in older adults. Sleep problems are also commonly associated with alcoholism. Yet few studies have examined the combined effects of alcoholism and aging on sleep. The purpose of this study was to investigate the main and interactive effects on sleep of diagnostic group and age group. METHOD: Alcohol dependent patients (n = 139; 118 men) and nonalcoholic controls (n = 87; 57 men) completed full-montage polysomnography, structured psychiatric diagnostic interviews, validated rating scales and alcohol histories at the University of Michigan between 1989 and 1996. The sample was divided for analytic purposes into older (aged 55+ years) and younger (aged <55 years) subgroups. RESULTS: After controlling for gender, race, body mass index and psychiatric-related sleep symptoms, alcoholics and older adults had significantly decreased total sleep time, and increased Stage 1 sleep percentage, respiratory distress and periodic limb movements. Older adults also had decreased delta sleep percentage and shorter rapid eye movement sleep latencies. Significant interactions were found between alcoholism and age group for Stage 1 sleep percentage, sleep-disordered breathing and periodic limb movements, with older alcoholics having the most disturbances. Older alcoholics had the highest mean values for sleep latency and the lowest mean values for sleep efficiency and delta sleep percentage when compared with the other three groups. CONCLUSIONS: Older alcoholics have increased sleep disturbances when compared with younger alcoholics and with nonalcoholics of both age groups. Care providers should screen for sleep problems among older adults with alcohol problems. PMID- 11414344 TI - MADD victim impact panels and stages-of-change in drunk-driving prevention. AB - OBJECTIVE: To assess the additional effects of Mothers Against Drunk Driving (MADD) Victim Impact Panels (VIPs) over the effects of a DWI (driving while intoxicated) school, on (1) moving individuals through the stages-of-change toward not drinking while driving and (2) drunk-driving recidivism. METHOD: A randomized experiment with 813 (75% male) DWI offenders in New Mexico measured progress through the stages-of-change at pretest, posttest, 1-year follow-up and 2-year follow-up. In addition, drunk-driving recidivism over 2 years was measured from state driving records. Individuals were randomly assigned to a DWI school or a DWI school plus a MADD VIP. RESULTS: No significant difference in movement through the stages-of-change, or in recidivism, occurred between respondents in the DWI-school-only treatment, and those in the DWI school plus VIP treatment. CONCLUSIONS: There was no additional effect of the MADD VIP, a relatively emotional intervention, over that of the DWI school, a relatively informational approach, on DWI behavior (whether measured by stages-of-change or by DWI rearrest data) over the 2-year period following the two interventions. PMID- 11414345 TI - The effect of an enhanced employee assistance program (EAP) intervention on EAP utilization. AB - OBJECTIVE: An enhanced employee assistance program (EAP) intervention was developed that delivers comprehensive EAP outreach services to all employees who may have alcohol-related and other workplace problems; standard EAP materials traditionally targeted at white men were enhanced to include women and minorities. This study evaluates whether the enhanced EAP intervention increased EAP utilization. METHOD: The enhanced EAP intervention was developed at a large community-based not-for-profit EAP located in Rockford, Illinois. Two primary worksites and 16 other newly contracted worksites received the enhanced EAP intervention and served as intervention sites; the 107 other worksites serviced by the EAP were used as comparison sites. We used time series data from 1991 to 1998 and included repeated measures on each firm's quarterly EAP utilization. RESULTS: The enhanced EAP intervention increased the mean number of women and minority cases per worksite by 58%, white male cases by 45% and total EAP cases by 53%. CONCLUSIONS: This study shows that, for a modest cost, the enhanced EAP intervention successfully increased utilization of EAP by all employees, especially utilization by women and minority employees. It also shows that traditional EAP services and outreach materials can be made more appealing to women and minorities without adversely affecting their utilization by white men. PMID- 11414346 TI - Prospective matching of alcoholic clients to cognitive-behavioral or interactional group therapy. AB - OBJECTIVE: In earlier work, client sociopathy and global psychopathology were effective variables for treatment matching: clients low on both sociopathy and severity of psychopathology were likely to benefit from interactional group therapy, whereas those scoring high on either of these dimensions benefited more from a coping skills intervention. The present study assessed whether outcomes improve further when clients are assigned to group treatments prospectively based on a matching strategy derived from the previous findings. METHOD: All participants (N = 250, 66% men) met criteria for alcohol dependence or abuse. About half were prospectively assigned to either cognitive-behavioral (CB) coping skills training or interactional therapy, those with higher levels of psychiatric severity or sociopathy were given CB and those who were low on both dimensions were given interactional therapy. The other half were randomly assigned to those treatments, replicating the procedure of the earlier study. Outcome data were collected at the conclusion of treatment and at 3-month intervals for 1 year following. RESULTS: Prospective matching of clients to treatment did not produce superior drinking outcomes compared to random treatment assignment. Randomly assigned clients were more likely to be abstinent at the end of treatment, but this effect disappeared at later follow-ups. Prospectively matched clients had fewer negative consequences of drinking than did those assigned randomly (unmatched). Neither sociopathy nor psychiatric severity was particularly effective for matching. CONCLUSIONS: The matching effects from our previous study were not replicated. Nevertheless, prospective matching did reduce the negative consequences of drinking, consistent with our previous results. PMID- 11414347 TI - Evaluating a brief alcohol intervention with fraternities. AB - OBJECTIVE: The current study tested the efficacy of a brief intervention designed to reduce drinking and drinking-related consequences among first-year fraternity members. METHOD: Twelve fraternities were randomly assigned to receive either a motivational enhancement intervention with individual and housewide feedback components (n = 6 houses) or a treatment-as-usual control condition (n = 6 houses). Individual feedback was delivered either by peer interviewers or professional research staff. Participants were assessed during their pledge (first) year of house membership and during a follow-up period 1 year later. RESULTS: Of the participants who completed follow-up (N = 120), fraternity members who received the brief intervention reported significant reductions in alcohol use (total average consumption) and typical peak blood alcohol concentrations when compared with fraternity members in the control condition. No differences in drinking-related consequences were observed. Fraternity members who received their individualized feedback from peer interviewers and professional members of the research staff reported similar outcomes. CONCLUSIONS: Results provide support for the efficacy of a brief motivational enhancement intervention in reducing drinking within this high-risk population. The cost-effective use of peer interviewers appears to be a promising strategy for delivering individualized prevention programming in college populations. PMID- 11414348 TI - Four-year outcomes from adolescent alcohol and drug treatment. AB - OBJECTIVE: Knowledge of treatment response for alcohol and drug problems among adults is mounting; less is known about long-term outcome for adolescents who receive treatment for alcohol and drug problems. The current study examined youth substance involvement over 4 years (using five waves of data collection) following treatment for alcohol and drug abuse. METHOD: A cohort of youth (N = 162, 60% male) treated during adolescence (mean age = 16 years) was followed into young adulthood, a period associated with stabilization of alcohol use patterns and elevated risk for life problems secondary to both alcohol and drug use. Participants (14-18 years old) were consecutive admissions to inpatient adolescent alcohol and drug treatment centers in San Diego that were abstinence focused and based on the 12-step approach. RESULTS: Alcohol and other drug use were reduced during the 4 years posttreatment, with the exception of nicotine. The greatest prevalence reduction occurred for stimulants; modest changes were evident in alcohol and marijuana use. Nicotine was the most commonly used substance throughout the 4 years after treatment. Several distinct substance involvement trajectories were evident during the 4 years following treatment. CONCLUSIONS: Alcohol and drug use patterns during the 4 years following treatment highlight both changes and diversity in substance involvement as youth make the transitions from middle to late adolescence and into young adulthood. Findings demonstrate the importance of identifying transitional periods and the need for alternative intervention strategies that may help the progression of this population into young adulthood. PMID- 11414349 TI - A randomized controlled trial of motivational enhancement therapy (MET) for mild to moderate alcohol dependence. AB - OBJECTIVE: This study was designed to conduct a randomized controlled trial of motivational enhancement therapy (MET) with two control conditions: nondirective reflective listening (NDRL) and no further counseling (NFC); and to conduct this study in a sample of patients with a primary diagnosis of mild to moderate alcohol dependence, in a "real-life" clinical setting. METHOD: Patients with mild to moderate alcohol dependence were recruited, assessed and treated at the Community Alcohol and Drug Service of Christchurch, New Zealand. All patients received a feedback/education session before randomization to either four sessions of MET, four sessions of NDRL, or NFC. Outcome data on 122 subjects (57.4% men) were obtained 6 months following the end of treatment, by an interviewer who was blind to the treatment condition. The primary drinking outcome was unequivocal heavy drinking, defined as drinking 10 or more standard drinks six or more times in the follow-up period. Global assessment scale (GAS) measured general personal/social functioning. RESULTS: Of patients treated with MET, 42.9% showed unequivocal heavy drinking compared with 62.5% of the NDRL and 65.0% of the NFC groups (p = .04). No significant differences were found for GAS score according to treatment condition. CONCLUSIONS: In patients with mild to moderate alcohol dependence, MET is more effective for reducing unequivocal heavy drinking than either a feedback/education session alone or four sessions of NDRL. MET can be considered an effective "value added" counseling intervention in a real-life clinical setting. In patients with mild to moderate alcohol dependence, nondirective reflective listening provides no additional advantage over a feedback/education session alone. PMID- 11414351 TI - Deliberations of the Task Force on Statistical Inference. PMID- 11414350 TI - Alcohol and eye diseases: a review of epidemiologic studies. AB - OBJECTIVE: Epidemiologic evidence for the associations of alcohol consumption with eye diseases was gathered. METHOD: A literature review was conducted. RESULTS: In addition to alcohol-induced ocular anomalies among children with fetal alcohol syndrome, epidemiologic studies in the past two decades have demonstrated that chronic alcoholism is associated with a significantly increased risk of cataract, keratitis, color vision deficiencies and corneal arcus. Moderate alcohol use, however, has been reported to be possibly protective against age-related macular degeneration, cataract and diabetic retinopathy. CONCLUSIONS: It is evident that through its biologic effects and effects on nutrient deficits alcohol plays a role in a variety of eye diseases. Information on the patient's drinking history can be clinically valuable and should be collected on a routine basis. PMID- 11414352 TI - CR1 and CR1-like: the primate immune adherence receptors. AB - Immune adherence describes the phenomenon in which complement-opsonized substrates, such as immune complexes (IC), viruses, or bacteria, are bound by primate erythrocytes via erythrocyte complement receptors. In vivo studies have shown that this binding allows the erythrocyte to act as an inert shuttle, targeting IC to the monocyte phagocytic system and away from vulnerable tissue. Thus, immune adherence appears to play an integral role in the primate in promoting the safe clearance of circulating IC and preventing IC-mediated pathologies. The complement receptors that mediate immune adherence comprise two unique but closely related gene products, either the type one complement receptor (CRI) in humans or CRI-like in non-human primates. This review focuses on the structure, function, and physiological role of the primate immune adherence receptors. PMID- 11414353 TI - Structure-function relationships of complement receptor type 1. AB - Human complement receptor type 1 (CR1) is a large, multifunctional glycoprotein which is a member of the regulators of complement activation family. Like other members of this family, it is composed mainly of tandemly arranged modules, each about 60-70 amino acids long, known as complement control protein repeats (CCPs). Each domain folds independently and contains a hydrophobic core wrapped in beta sheets. These domains mediate interactions with C3/C4-derived fragments. CR1 is the most versatile inhibitor of both classical and alternative pathway C3 and C5 convertases due to its decay-accelerating activity and co-factor activity for C3b/C4b cleavage. Moreover, CR1 plays a major role in immune complex clearance due to its high affinity for C3b and C4b. CR1 is an excellent model to study structure-function relationships because its functions are mediated by two distinct but highly homologous sites, each composed of three CCPs. CR1 derivatives carrying just one active site were used to define critical sequences/amino acids. This was achieved by testing functional profiles of the proteins carrying a mutated active site produced by substituting peptides/amino acids with their counterparts from the other site. These mutated proteins, of which we analyzed over 100, permitted the fine mapping of the functional sites. CR1 on primate erythrocytes varies in size. In most cases it is smaller and has fewer active sites than does human CR1. This variation was used to determine that increased copy number (3,000 to 20,000 versus 300 for human CR1) compensates for a smaller size. Moreover, studies of primate CR1 led to the finding that subtle differences in the critical areas, as compared to human sites, produce active sites with a broader functional repertoire. These alterations ensure that short CR1 forms possess similar biologic activities to the large CR1 forms. There is much interest in producing therapeutic agents to inhibit unwanted complement activation. Based on these structure-function analyses, smaller and more potent complement inhibitors derived from CR1 can be produced. PMID- 11414354 TI - Structural biology of the alternative pathway convertase. AB - Complement convertases are bimolecular complexes expressing protease activity only against C3 and C5. Their action is necessary for production of the biological activities of the complement system. Formation of these complexes proceeds through sequential protein-protein interactions and proteolytic cleavages of high specificity. Recent structural, mutational and functional data on factors D and B have significantly enhanced our understanding of the assembly, action, and regulation of the alternative pathway convertase. These processes were shown to depend critically on conformational changes, only some of which are reversible. The need for such changes is dictated by the zymogen-like configurations of the active centers of these unique serine proteases. The structural determinants of some of these changes have been defined from structural and mutational analyses of the two enzymes. Transition of factor D from the zymogen-like to the catalytically active conformation is completely reversible, while the active conformation of the catalytic center of the Bb fragment of factor B is irreversibly attenuated to a great extent on dissociation of the convertase complex. Both mechanisms contribute to the regulation of the proteolytic activity of these enzymes. Additional studies are necessary for a complete description of the elegant mechanisms mediating these processes. PMID- 11414355 TI - Structural biology of C1: dissection of a complex molecular machinery. AB - The classical pathway of complement is initiated by the C1 complex, a multimolecular protease comprising a recognition subunit (C1q) and two modular serine proteases (C1r and C1s) associated as a Ca2+-dependent tetramer (C1s-C1r C1r-C1s). Early studies have allowed identification of specialized functional domains in these proteins and have led to low-resolution models of the C1 complex. The objective of current studies is to gain deeper insights into the structure of C1, and the strategy used for this purpose mainly consists of dissecting the C1 components into modular fragments, in order to solve their three-dimensional structure and establish the structural correlates of their function. The aim of this article is to provide an overview of the structural and functional information generated by this approach, with particular emphasis on the domains involved in the assembly, the recognition function, and the highly specific proteolytic properties of C1. PMID- 11414356 TI - Structure and flexibility of the multiple domain proteins that regulate complement activation. AB - In this review we summarise more than 10 years of biophysical exploration into the structural biology of the regulators of complement activation (RCA). The five human proteins responsible for regulation of the early events of complement are homologous and are composed largely from building blocks called "complement control protein (CCP) modules". Unlike most multiple domain proteins they do not contain any of the other widely occurring module types. This apparent simplicity of RCA structure, however, is belied by their sophistication of function. In fact, the structures of the individual CCP modules exhibit wide variations on a common theme while the extent and nature of intermodular connections is diverse. Some neighbouring modules within a protein stabilise each other and some co operate to form specific binding surfaces. The degree of true "modularity" of CCPs is open to debate. The study of RCA proteins clearly illustrates the value of combining complementary structural biology techniques. The results could have implications for folding, evolution, flexibility and structure-function relationships of other molecules in the large, diverse and little understood category of multiple domain proteins. PMID- 11414357 TI - The ancestral complement system in sea urchins. AB - The origin of adaptive immunity in the vertebrates can be traced to the appearance of the ancestral RAG genes in the ancestral jawed vertebrate; however, the innate immune system is more ancient. A central subsystem within innate immunity is the complement system, which has been identified throughout and seems to be restricted to the deuterostomes. The evolutionary history of complement can be traced from the sea urchins (members of the echinoderm phylum), which have a simplified system homologous to the alternative pathway, through the agnathans (hagfish and lamprey) and the elasmobranchs (sharks and rays) to the teleosts (bony fish) and tetrapods, with increases in the numbers of complement components and duplications in complement pathways. Increasing complexity in the complement system parallels increasing complexity in the deuterostome animals. This review focuses on the simplest of the complement systems that is present in the sea urchin. Two components have been identified that show significant homology to vertebrate C3 and factor B (Bf), called SpC3 and SpBf, respectively. Sequence analysis from both molecules reveals their ancestral characteristics. Immune challenge of sea urchins indicates that SpC3 is inducible and is present in coelomic fluid (the body fluids) in relatively high concentrations, while SpBf expression is constitutive and is present in much lower concentrations. Opsonization of foreign cells and particles followed by augmented uptake by phagocytic coelomocytes appears to be a central function for this simpler complement system and important for host defense in the sea urchin. These activities are similar to some of the functions of the homologous proteins in the vertebrate complement system. The selective advantage for the ancestral deuterostome may have been the amplification feedback loop that is still of central importance in the alternative pathway of complement in higher vertebrates. Feedback loop functions would quickly coat pathogens with complement leading to phagocytosis and removal of foreign cells, a system that would be significantly more effective than an opsonin that binds upon contact as a result of simple diffusion. An understanding of the immune response of the sea urchin, an animal that is a good estimator of what the ancestral deuterostome immune system was like, will aid us in understanding how adaptive immunity might have been selected for during the early evolution of the vertebrates and how it might have been integrated into the pre-existing innate immune system that was already in place in those animals. PMID- 11414358 TI - Carboxypeptidase R is an inactivator of complement-derived inflammatory peptides and an inhibitor of fibrinolysis. AB - Carboxypeptidase R (CPR) exists in precursor form (proCPR) in plasma in contrast to carboxypeptidase N (CPN), which is present in the active state. CPR plays two important roles, one of which appears to be the control of the inflammatory response by inactivation of anaphylatoxins such as complement-derived C3a and C5a. Therefore, an increase in CPR activity may facilitate rapid inactivation of these inflammatory mediators generated at the site of bacterial infection. Upregulation of proCPR expression during the inflammatory response initiated for instance by endotoxin (lipopolysaccharide) should play a role in suppressing hyper-reactivity as seen in septic shock. CPR also functions as an inhibitor of fibrinolysis, where its ability to prevent binding of plasminogen to lysine residues on fibrin clots significantly lengthens tissue plasminogen activator (tPA)-induced fibrinolysis time. Therefore, upregulation of proCPR production during the inflammatory response may exacerbate thrombosis contributing to the development of disseminated intravascular coagulation as well as other conditions involving thrombosis. Co-administration of tPA and a specific inhibitor of CPR, such as potato carboxypeptidase inhibitor, which does not affect CPN, may be useful in thrombolytic therapy. PMID- 11414359 TI - The supportive role of complement in HIV pathogenesis. AB - This review focuses on interactions of HIV with the first-line defence of native immunity, the complement system. In all body compartments tested so far, HIV meets complement. Activation of the complement system results in deposition of C3 fragments on the viral surface, but in contrast to other pathogens, most of HIV is not or is only poorly lysed by membrane attack complexes. To survive complement-mediated lysis, HIV has not only developed resistance mechanisms, but uses opsonisation with complement fragments for its own advantage. Opsonised virions interact with complement receptor-expressing cells, which are either subsequently infected with high efficiency or retain viral particles on their surface, which promotes transmission of virus to other permissive cells. Our knowledge of these mechanisms has increased enormously over the past few years. A complete understanding of these complex interactions of HIV with the complement system opens new perspectives for development of alternative therapeutic strategies. PMID- 11414360 TI - Targeting complement in therapy. AB - With increasing evidence that complement activation significantly contributes to the pathogenesis of a large number of inflammatory diseases, strategies that interfere with its deleterious action have become a major focus in pharmacological research. Endogenous soluble complement inhibitors (C1 inhibitor, recombinant soluble complement receptor 1, antibodies) blocking key proteins of the cascade reaction, neutralizing the action of the complement-derived anaphylatoxin C5a, or interfering with complement receptor 3 (CR3, CD18/11b) mediated adhesion of inflammatory cells to the vascular endothelium have successfully been tested in various animal models over the past years. Promising results consequently led to clinical trials. Furthermore, incorporation of membrane-bound complement regulators (decay-accelerating factor (CD55), membrane co-factor protein (CD46), CD59) in transgenic animals has provided a major step forward in protecting xenografts from hyperacute rejection. At the same time, the poor contribution of complement to the antitumor response, which is caused by multiple resistance mechanisms that hamper the efficacy of antibody-based tumor therapy, is increasingly recognized and requires pharmacologic intervention. First attempts have now been made to interfere with the resistance mechanisms, thereby improving complement-mediated tumor cell destruction. PMID- 11414361 TI - Structure and biology of complement protein C3, a connecting link between innate and acquired immunity. AB - Complement protein C3 is a central molecule in the complement system whose activation is essential for all the important functions performed by this system. After four decades of research it is now well established that C3 functions like a double-edged sword: on the one hand it promotes phagocytosis, supports local inflammatory responses against pathogens, and instructs the adaptive immune response to select the appropriate antigens for a humoral response; on the other hand its unregulated activation leads to host cell damage. In addition, its interactions with the proteins of foreign pathogens may provide a mechanism by which these microorganisms evade complement attack. Therefore, a clear knowledge of the molecule and its interactions at the molecular level not only may allow the rational design of molecular adjuvants but may also lead to the development of complement inhibitors and new therapeutic agents against infectious diseases. PMID- 11414362 TI - Role of the C5b-9 complement complex in cell cycle and apoptosis. AB - Assembly of C5b-9 on cell membranes results in transmembrane channels and causes cell death. When the number of C5b-9 molecules is limited, nucleated cells are able to escape cell death by endocytosis and by shedding of membranes bearing C5b 9. Sublytic CSb-9 induces proto-oncogenes, activates the cell cycle, and enhances cell survival. In addition, C5b-9 reverses the differentiated phenotype of postmitotic cells, such as oligodendrocytes and skeletal muscle cells. The signal transduction pathways responsible for cell cycle activation by CSb-9 include Gi mediated activation of extracellular signal-regulated kinase 1 and phosphatidylinositol 3-kinase (PI3-K). Cell survival enhanced by C5b-9 is mediated by the PI3-K/Akt pathway, which inhibits apoptosis through regulation of BAD. These findings indicate that complement activation and membrane assembly of sublytic C5b-9 play an important role in inflammation by promoting cell proliferation and by rescuing apoptotic cells. PMID- 11414363 TI - The role of complement in inflammation and adaptive immunity. AB - Major advances in our understanding of the immunobiology of complement were made within the past 5 years primarily due to the development of gene-targeting technology. New strains of mice bearing specific deficiencies in serum complement proteins or their receptors were developed using this approach. Characterization of these mice has provided new and exciting insights into the biology of the complement system. In this review, we discuss recent results on two important aspects of the complement system, i) host protection and inflammation, and ii) regulation of B lymphocytes of adaptive immunity. While these two roles appear distinct, they are linked. We discuss how natural antibody and classical pathway complement work together in host protection against bacterial infection on the one hand but, on the other, they co-operate to induce inflammation as observed in reperfusion injury. Significantly, the lymphocytes that produce natural antibody, the B-1 lymphocytes, are regulated in part by the complement system. PMID- 11414364 TI - Human blood platelet gC1qR/p33. AB - Platelets are involved in the development of many types of vascular lesions. In addition to their role in primary hemostasis, they participate in inflammatory processes that may contribute to the development of thrombosis, atherosclerosis and vasculitis. In this regard, we have been interested in platelet interactions with the complement subcomponent C1q. C1q has been shown to modulate platelet interactions with collagen and immune complexes, and has been identified at sites of vascular injury and inflammation, as well as in atherosclerotic lesions. Platelets express a variety of C1q binding sites, including gC1qR/p33 (gC1qR), a multifunctional, multicompartment cellular protein. Here we focus on the structure and function of platelet gC1qR and its emerging role in modulating platelet function at sites of vascular injury and inflammation. PMID- 11414365 TI - gC1q-R/p33, a member of a new class of multifunctional and multicompartmental cellular proteins, is involved in inflammation and infection. AB - Human gC1q-R (p33, p32, C1qBP, TAP) is a ubiquitously expressed, multiligand binding, multicompartmental cellular protein involved in various ligand-mediated cellular responses. Although expressed on the surface of cells, an intriguing feature of the membrane-associated form of gC1q-R is that its translated amino acid sequence does not predict the presence of either a sequence motif compatible with a transmembrane segment or a consensus site for a glycosylphosphatidylinositol anchor. Moreover, the N-terminal sequence of the pre pro-protein gC1q-R contains a motif that targets the molecule to the mitochondria and as such was deemed unlikely to be expressed on the surface. However, several lines of experimental evidence clearly show that gC1q-R is present in all compartments of the cell, including the extracellular cell surface. First, surface labeling of B lymphocytes with the membrane-impermeable reagent sulfosuccinimidyl 6-(biotinamido)hexanoate shows specific biotin incorporation into the surface-expressed but not the intracellular form of gC1q-R. Second, FACS and confocal laser scanning microscopic analyses using anti-gC1q-R IgG mAb 60.11 or 74.5.2, and the fluorophore Alexa 488-conjugated F(ab')2 goat anti-mouse IgG as a probe, demonstrated specific staining of Raji cells (>95% viable). Three dimensional analyses of the same cells by confocal microscopy showed staining distribution that was consistent with surface expression. Third, endothelial gC1q R, which is associated with the urokinase plasminogen activator receptor, and cytokeratin 1 bind 125I-high molecular weight kininogen in a specific manner, and the binding is inhibited dose-dependently by mAb 74.5.2 recognizing gC1q-R residues 204-218. Fourth, native gC1q-R purified from Raji cell membranes but not intracellular gC1q-R is glycosylated, as evidenced by a positive periodic acid Schiff stain as well as sensitivity to digestion with endoglycosidase H and F. Finally, cross-linking experiments using C1q as a ligand indicate that both cC1q R and gC1q-R are co-immunoprecipitated with anti-C1q. Taken together, the evidence accumulated to date supports the concept that in addition to its intracellular localization, gC1q-R is expressed on the cell surface and can serve as a binding site for plasma and microbial proteins, but also challenges the existing paradigm that mitochondrial proteins never leave their designated compartment. It is therefore proposed that gC1q-R belongs to a growing list of a class of proteins initially targeted to the mitochondria but then exported to different compartments of the cell through specific mechanisms which have yet to be identified. The designation 'multifunctional and multicompartmental cellular proteins' is proposed for this class of proteins. PMID- 11414366 TI - Ficolins and the lectin complement pathway. AB - Ficolins, found in various tissues, are a group of proteins containing both a collagen-like and a fibrinogen-like domain. Recently, it was shown that ficolins present in serum are lectins with a common binding specificity for N acetylglucosamine (GlcNAc). The fibrinogen-like domain is responsible for the carbohydrate binding. Mannose-binding lectin (MBL) is also a collagenous lectin in serum that is specific for GlcNAc and mannose binding. Its domain organization is similar to that of ficolins, except that MBL has a carbohydrate-recognition domain instead of a fibrinogen-like domain. MBL plays a role in innate immunity by acting as an opsonin and activating complement in association with MBL associated serine protease (MASP) via the lectin pathway. Investigations of two types of human serum ficolins, ficolin/P35 and Hakata antigen, revealed that they are associated with MASPs and sMAP, a truncated protein of MASP-2, and that they activate complement. These findings indicate that serum ficolins are structurally and functionally similar to MBL and have the capacity to activate the lectin pathway and thus have a role in innate immunity. PMID- 11414367 TI - Mannose-binding lectin: targeting the microbial world for complement attack and opsonophagocytosis. AB - Mannose-binding lectin (MBL) is an important constituent of the innate immune system. This protein binds through multiple lectin domains to the repeating sugar arrays that decorate many microbial surfaces, and is then able to activate the complement system through a specific protease called MBL-associated protease-2. We have used flow cytometry to study both the binding of MBL to microorganisms and the subsequent activation of complement. For selected Gram-negative organisms, such as Salmonella and Neisseria, we have examined the relative roles of lipopolysaccharide (LPS) structure and capsule in determining binding and conclude that the LPS is of major importance. Our results from studies with several clinically relevant organisms also show that MBL binding detected by flow cytometry leads to measurable activation of purified C4, suggesting that the bound lectin is capable of initiating opsonophagocytosis and/or bacterial lysis. There is an increasing literature suggesting that MBL deficiency, which mainly results from three relatively common single point mutations in exon 1 of the gene, predisposes both to infection by extracellular pathogens and to autoimmune disease. In addition, the protein also modulates disease severity, at least in part through a complex, dose-dependent influence on cytokine production. The mechanisms and signalling pathways involved in such processes remain to be elucidated. PMID- 11414368 TI - Toward a structure- and process-integrated view of personality: traits as density distribution of states. AB - Three experience-sampling studies explored the distributions of Big-Five-relevant states (behavior) across 2 to 3 weeks of everyday life. Within-person variability was high, such that the typical individual regularly and routinely manifested nearly all levels of all traits in his or her everyday behavior. Second, individual differences in central tendencies of behavioral distributions were almost perfectly stable. Third, amount of behavioral variability (and skew and kurtosis) were revealed as stable individual differences. Finally, amount of within-person variability in extraversion was shown to reflect individual differences in reactivity to extraversion-relevant situational cues. Thus, decontextualized and noncontingent Big-Five content is highly useful for descriptions of individuals' density distributions as wholes. Simultaneously, contextualized and contingent personality units (e.g., conditional traits, goals) are needed for describing the considerable within-person variation. PMID- 11414369 TI - The heritability of attitudes: a study of twins. AB - The genetic basis of individual differences in attitudes was examined in a survey of 195 pairs of monozygotic twins and 141 pairs of same-sex dizygotic twins. A principal components analysis of the 30 attitude items in the survey identified 9 attitude factors, of which 6 yielded significant heritability coefficients. Nonshared environmental factors accounted for the most variance in the attitude factors. Possible mediators of attitude heritability were also assessed, including personality traits, physical characteristics, and academic achievement. Analyses showed that several of these possible mediators correlated at a genetic level with the heritable attitude factors, suggesting that the heritability of the mediator variables might account for part of the heritable components of some attitudes. There was also some evidence that highly heritable attitudes were psychologically "stronger" than less heritable attitudes. PMID- 11414370 TI - To do or not to do: desirability and consistency mediate judgements of regret. AB - In 4 studies, the authors demonstrated that when errors associated with action were inconsistent with decision nakers' orientation, they were undesirable and produced more regret than did errors associated with inaction. Conversely, when errors associated with action were consistent with decision makers' orientation. they were desirable and produced less regret than did errors associated with inaction. Desirability and consistency mediated this relationship, independent of mutability. These results were obtained when judgments and decisions to act or not act were made in close temporal proximity to one another as well as when participants reflected on their past decisions. The authors provide an analysis of when counterfactuals would and would not be expected to mediate judgments of normality and regret. PMID- 11414371 TI - That "vision thing": the state of theory in social and personality psychology at the edge of the new millennium. AB - Social psychololgy's status as a theoretical discipline is assessed. Whereas it has excelled as an experimental science, the field has generally eschewed broad theorizing and tended to limit its conceptualizations to relatively narrow, "mid range" notions closely linked to the operational level of analysis. Such "theory shyness" may have spawned several negative consequences, including the tendency to invent new names for old concepts, fragmentation of the field, and isolation from the general cultural dialogue. Recently, steps have been taken to encourage greater theoretical activity by social psychologists, and there are now several major outlets for theoretical contributions. Further initiatives are needed, however, to instigate theoretical creativity, including ways of overcoming disciplinary risk aversion and the training of young social psychologists in ways and means of theory construction. PMID- 11414372 TI - Person theories and attention allocation: preferences for stereotypic versus counterstereotypic information. AB - How do people respond to information that counters a stereotype? Do they approach it or avoid it? Four experiments showed that attention to stereotype-consistent vs. -inconsistent information depends on people's implicit theories about human traits. Those holding an entity theory (the belief that traits are fixed) consistently displayed greater attention to (Experiments 1 and 4) and recognition of (Experiments 2 and 3) consistent information. whereas those holding an incremental (dynamic) theory tended to display greater attention to (Experiment 1) and recognition of (Experiment 3) inconsistent information. This was true whether implicit theories were measured as chronic structures (Experiments 1, 2, and 4) or were experimentally manipulated (Experiment 3). Thus, different a priori assumptions about human traits and behavior lead to processing that supports versus limits stereotype maintenance. PMID- 11414373 TI - Human mate poaching: tactics and tempations for infiltrating existing mateships. AB - The authors explored the psychology of romantically attracting someone who is already in a relationship--what can be called the process of human mate poaching. In Study 1 (N = 236), they found that attempts at poaching were relatively common and were linked with distinctive personality dispositions. Study 2 (N = 220) documented that the perceived costs and benefits of poaching differed somewhat for men and women and depended on whether short-term or long-term poaching outcomes were targeted. Study 3 (N = 453) found support for 5 evolution-based hypotheses about the perceived effectiveness of poaching tactics. Study 4 (N = 333) found that poaching effectiveness was influenced by the type of relationship being encroached on-marital, dating, long distance, highly committed, just beginning, or about to end. Discussion focuses on the importance of placing mate poaching within the broader context of human sexual strategies. PMID- 11414374 TI - Quality of decision making and group norms. AB - Two studies investigated the impact of group norms for maintaining consensus versus norms for critical thought on group decisions in a modification of the biased sampling paradigm (G. Stasser & W. Titus, 1985). Both studies showed that critical norms improved the quality of decisions, whereas consensus norms did not. This effect appeared to be mediated by the perceived value of shared and unshared information: Consensus norm groups valued shared information more highly than critical groups did, and valence was a good predictor of decision outcome. In addition, the 2nd study showed that the group norm manipulation has no impact on individual decisions, consistent with the assumption that this is a group effect. Results suggest that the content of group norms is an important factor influencing the quality of group decision-making processes and that the content of group norms may be related to the group's proneness for groupthink. PMID- 11414375 TI - Uncertainty management: the influence of uncertainty salience on reactions to perceived procedural fairness. AB - On the basis of fairness heuristic theory, it is argued in this article that people especially need fairness when they are reminded about aspects of their lives that make them uncertain. It is therefore proposed that thinking about uncertainty should make fairness a more important issue to people. The findings of 3 experiments support this line of reasoning: Asking (vs. not asking) participants 2 questions that solicited their thoughts and feelings of being uncertain led to stronger effects of perceived procedural fairness on participants' affective reactions toward the way they were treated. It is argued that these findings suggest that fairness matters to people especially when they are trying to deal with things that make them uncertain. An implication of the current findings therefore may be that fairness is important to people because it gives them an opportunity to manage uncertain aspects of their lives. PMID- 11414376 TI - Battle of the sexes: gender stereotype confirmation and reactance in negotiations. AB - The authors examined how gender stereotypes affect negotiation performance. Men outperformed women when the negotiation was perceived as diagnostic of ability (Experiment 1) or the negotiation was linked to gender-specific traits (Experiment 2), suggesting the threat of negative stereotype confirmation hurt women's performance relative to men. The authors hypothesized that men and women confirm gender stereotypes when they are activated implicitly, but when stereotypes are explicitly activated, people exhibit stereotype reactance, or the tendency to behave in a manner inconsistent with a stereotype. Experiment 3 confirmed this hypothesis. In Experiment 4, the authors examined the cognitive processes involved in stereotype reactance and the conditions under which cooperative behaviors between men and women can be promoted at the bargaining table (by activating a shared identity that transcends gender). PMID- 11414377 TI - Money and subjective well-being: it's not the money, it's the motives. AB - Past researchers have argued that the relative importance a person attaches to money is negatively related to subjective well-being (SWB). However, the past research suffers from the theoretical problem of not including the diferent motives for making money. With a sample of 240 business students, the authors developed a set of scales to measure motives for making money. They used a sample of 492 business students to confirm the factor structure of these motives. With another sample of 266 business students, the authors found that the negative relationship between money importance and SWB was due to the two variables being the result of a common cause; namely, the motives of social comparison. seeking power, showing off, and overcoming self-doubt. This finding was replicated with a sample of 145 entrepreneurs. PMID- 11414378 TI - Predictors of caregiving in adult intimate relationships: an attachment theoretical perspective. AB - Romantic couples (N = 194) participated in an investigation of caregiving processes in adulthood. In Phase 1, couple members completed questionnaires designed to identify attachment style differences in caregiving behavior and to explore the underlying (personal and relationship) mechanisms that lead people with different attachment styles to be effective or ineffective caregivers. Results revealed that social support knowledge, prosocial orientation. interdependence, trust, and egoistic motivation mediated the link between attachment style and caregiving. In Phase 2, responsive caregiving was assessed behaviorally by exposing one member of the couple to a stressful laboratory situation and experimentally manipulating his or her need for support. Results revealed that attachment style and mediating mechanisms identified in Phase 1 also predicted observable support behavior in a specific episode in which a partner had a clear need for support. PMID- 11414379 TI - Three paths of adult development: conservers, seekers, and achievers. AB - This study examined the development of individuals whose motivations and skills led them to develop in different but equally positive ways. C. D. Ryff's (1989) scales for Environmental Mastery (EM) and Personal Growth (PG) were used to identify three configurations of positive mental health in 111 women of the Mills Longitudinal Study: Achievers, high on both scales; Conservers, high on EM, low on PG; and Seekers, high on PG, low on EM. Each pattern showed a distinctive profile of strengths on four criteria of maturity--competence, generativity, ego development, and wisdom--and each was predicted by distinctive features of positive and negative emotionality, identity processes, and change in self control across 31 years of adulthood. Identity at age 43 mediated the influence of personality at age 21 in predicting positive mental health pattern at age 60. PMID- 11414380 TI - Resistance to ciprofloxacin and the new fluoroquinolones. AB - Resistance to ciprofloxacin has generally remained low in the UK. A low rate of resistance development in vitro has been suggested for the newer fluoroquinolones such as grepafloxacin and trovafloxacin. Levofloxacin is likely to have a pattern of development of resistance similar to that of ofloxacin. Resistance problems with ofloxacin are being reported in the USA, but it is unclear whether similar problems will arise with levofloxacin. Restrained quinolone use, surveillance of pathogen susceptibility and control of cross-infection with resistant clones remain important strategies for managing fluoroquinolone resistance. PMID- 11414381 TI - Cost and dosing issues. AB - Twice-daily dosing is recommended for ciprofloxacin for most indications. Trovafloxacin and grepafloxacin can be administered once-daily. Levofloxacin should probably be administered twice daily in serious, life-threatening infections. The acquisition cost of ciprofloxacin is currently higher than the newer agents, but a lower acquisition cost does not necessarily mean a lower overall cost of treatment. A cheaper agent which is less effective or safe just shifts costs to other areas by creating further clinical problems with their attendant costs. The total costs associated with treatment with newer agents are as yet undefined. Restricting the prescription of a particular antibiotic may result in the increased use of an alternative regimen which may cost the same or even more than the original ('cost shifting'). This interesting observation needs to be substantiated with future research. Ciprofloxacin is available in both intravenous and oral formulations and suitable for sequential intravenous-oral therapy with its attendant benefits. It is currently the gold-standard quinolone in the hospital setting for a broad range of infections, particularly Pseudomonas aeruginosa infections. Initially, the newer quinolones are most likely to be used to treat severe, community-acquired pneumonia. PMID- 11414382 TI - Ciprofloxacin: efficacy and indications. AB - Ciprofloxacin's strength is in 'below-the-diaphragm' indications. Information about the efficacy of the new fluoroquinolones in this indication is currently limited. Ciprofloxacin is effective in both uncomplicated and complicated urinary tract infections, but is probably best reserved for complicated, hospital acquired or recurrent infections. Ciprofloxacin is effective in complicated and severe lower respiratory tract infections, including those in patients with infective exacerbations of chronic bronchitis and cystic fibrosis, and pseudomonal infections. Ciprofloxacin is effective in the treatment of serious, non-self-limiting intra-abdominal infections, peritonitis in CAPD, pelvic inflammatory disease, endometritis and gall-bladder infections. Ciprofloxacin is effective in the treatment of a range of serious, non-self-limiting gastrointestinal infections (e.g. Salmonella and typhoid fever). Ciprofloxacin is effective in the empirical treatment of febrile neutropenic episodes, and prophylaxis of gram-negative bacteraemia in neutropenia and bone marrow transplantation. Ciprofloxacin is also effective in a range of other indications (e.g. eye infections, skin and soft tissue infections, bone and joint infections, and gonorrhoea). PMID- 11414383 TI - Safety of the new fluoroquinolones compared with ciprofloxacin. AB - Significant safety issues that have arisen with fluoroquinolones include phototoxicity, cardiotoxicity, tendinitis, CNS effects and drug interactions. Ciprofloxacin is well tolerated; the incidence of adverse events is low and serious adverse events are rare. Levofloxacin has a reduced CNS adverse event rate compared with ofloxacin. Sparfloxacin has significant phototoxicity and potential cardiac toxicity. Grepafloxacin has significantly increased adverse event rates, particularly gastrointestinal intolerance. Taste perversion and nausea are common. Trovafloxacin has an increased potential for CNS adverse reactions, notably dizziness. Post-marketing surveillance data indicate the possibility of serious hepatic reactions and pancreatitis. Interactions between fluoroquinolones and drugs metabolised by the hepatic cytochrome P450 system affect the clearance of theophylline and caffeine. Quinolone absorption is significantly reduced by co-administration of antacids. Hospitalised patients are likely to be receiving multiple-drug therapy, but drug interactions are avoidable. The interactions of specific fluoroquinolones should be checked prior to prescription. PMID- 11414384 TI - Anthropological perspectives on alcohol and drugs at the turn of the new millennium. AB - This introduction to the collection provides our thoughts on where alcohol and drug studies in anthropology are going as we enter the new millennium. After commenting briefly on each of the papers that comprise the rest of the volume, we discuss what we see as the most important and exciting issues in the future and give our views on what alcohol and drug studies can offer to medical anthropology, anthropology writ large, interdisciplinary and multidisciplinary research, and the realm of public policy and practical affairs. We call for a continued study by anthropologists of the whole array of pharmacologically active substances used by humans in different parts of the world, whether or not such studies are situated within medical anthropology. We note that many of these substances have received little attention from anthropologists to date, quite strikingly so in the cases of substances such as marijuana and methamphetamines. We emphasize that most scholars working in the anthropology of alcohol and drugs are concerned with the application of their findings to social problems, and we note that this has been especially true of research on alcoholic beverages and injection drugs. This leads us to a discussion of anthropology's involvement in public health intervention and policy work in a variety of settings. Such involvement is shown to have informed anthropological theory (notably political economic approaches) and to have enriched the methodological toolkits and forms of data analysis anthropologists use. Perhaps more importantly, we argue that such multidisciplinary involvement in applied work is most likely to eventuate in theoretical progress in alcohol and drug studies, since theory in the social sciences is not bound to singular disciplinary approaches. Thus we advocate for a "hybrid vigor" in this specialty area in the years ahead. PMID- 11414385 TI - Socio-cultural anthropology and alcohol and drug research: towards a unified theory. AB - The arrival of the millennium has understandably created a growing tendency for social commentators to evaluate the current state of the world, assess prior developments and suggest new and enlightened ways forward. In this time of re appraisals, we assess the current state of the anthropology of alcohol and drug research, consider its early history, examine the range and theoretical underpinnings of work done today, and propose the elements of a possible future model. In formulating the model, we have borrowed from a number of different theoretical approaches and insights not only from anthropological discussions of alcohol and drug issues but also from anthropological research outside these arenas. In examining these issues, and as a way of attempting to answer the question--where has the field come from and where is it heading--we situate anthropological contributions to this area not merely within the wider context of anthropological thought and research, but more importantly within a wider social and political context that takes account of the organizational, funding and conceptual influences, constraints and pressures that operate on anthropologists who wish to conduct research on alcohol and drug issues. PMID- 11414386 TI - Intoxication and bad behaviour: understanding cultural differences in the link. AB - Research developments since the appearance of MacAndrew and Edgerton's landmark volume, Drunken Comportment (1969), are summarized. The challenge of moving beyond the book is to understand what lies behind cultural variations in drunken comportment. Four specific factors in variations in drunken comportment are discussed. (1) A common contrast is between "wet" societies, where drinking is banalized everyday, and "dry" societies, where alcohol is set apart as a special commodity. Problems with this contrast are discussed, and the need for cross cultural studies comparing expectancies from intoxication. (2) There is a need to study variations in the definition of intoxication as a "time out" state. In some societies, intoxication is likened to possession by spirits; a rationalistic version of this can be found in Canadian court decisions viewing extreme intoxication as potentially "akin to automatism". (3) If bad behaviour is a foreseeable consequence of drinking, why do some societies nevertheless not hold the drinker responsible'? In Anglo-American and similar societies, drunkenness has some excuse value, but it is not a very good excuse. Compromises like this seem to be found also in other cultures. (4) Pseudointoxication is fairly widespread, and seems to mark social situations where alcohol has enhanced excuse value. It appears to be a stratagem of the weaker side across cultural boundaries, and of the young where age-grading favours older groups. Concerning the possibility of cultural changes in drunken comportment, it is argued that there are historical examples, but such a shift requires a substantial social change. PMID- 11414387 TI - Toward a bio-cultural and political economic integration of alcohol, tobacco and drug studies in the coming century. AB - The 18th and early 20th centuries witnessed the disintegration of a unified approach to understanding the human condition. Political economy, the broad study of human society, fragmented into an array of university-based disciplines, each reductionistically focused on its narrow arena of specialized research. Medicine, which had been concerned with health in social and historic contexts, narrowed its focus to the microscopic level and to encapsulated understandings of the immediate effects of pathogens and of the structures of disintegrated organ systems. Similarly, anthropology, which continued to wave a banner of holism, retreated for much of the 20th century into fine-grained cultural studies of seemingly isolated human communities on the one hand, and highly specialized biological and biobehavioral analyses of only tangential concern to cultural concerns on the other. Consequently, it has appeared at times as if anthropology would fragment into two or more disciplines and the opportunity for an integrated understanding of the human condition would be lost in the process. As we approach ever closer to the 21st century, however, the felt need for interdisciplinary and intradisciplinary reintegration has grown stronger. This trend is manifest increasingly in the field of alcohol, tobacco, and drug studies, and suggests one of the places anthropology may be going in the future. In this light, this paper examines the use of a critical biocultural model employed in the anthropological assessment of the Hartford Syringe Exchange Program. This model integrates the political economy of risk behavior, the ethnographic examination of insider understandings, meaning systems and behaviors, and the biological analysis of health-related issues. Methodologically, the assessment combined methods and concepts from all of the major subfields of anthropology. PMID- 11414388 TI - Assessing alcohol consumption: developments from qualitative research methods. AB - This paper examines some shortcomings of self-report questionnaires used to assess alcohol use quantity and frequency and demonstrates the advantages of qualitative interviews to more accurately capture drinking patterns among adolescents. The paper considers alcohol use among two ethnic groups of Black adolescents and discusses variations in rates of alcohol consumption. Qualitative interview data collected from African-American and Haitian adolescents suggest higher rates of alcohol consumption than indicated in previous studies of Black adolescents. Furthermore, qualitative interview data demonstrate greater differences across groups in drinking patterns, including amount and type of alcohol consumed in different cultural contexts than have quantitative-based studies. Strategies are suggested for improving the methodology used to assess drinking patterns among adolescents. PMID- 11414389 TI - Culture and the restoration of self among former American Indian drinkers. AB - This paper explores the social and cultural context of remission from alcohol problems in an urban American Indian community. Using the discourse of interviews conducted with 48 self-defined problem drinkers, 13 of whom had abstained from alcohol for at least a year prior to the interview, it explores the ways in which alcohol problems have been understood and dealt with by these Indian men and women. Drawing on the ethnographic literature on AA and culturally specific healing practices, the analysis centers on how new understandings of the self are articulated in sobriety and the ways in which this discourse draws on the themes of cultural restoration that are widely articulated in Indian communities. The paper closes with a consideration of how this inquiry with a community sample forces us to broaden models of self transformation that are derived from work in more circumscribed institutional contexts and, in turn, how the testimony of these men and women forces us to take quite seriously the idea that cultural restoration can be crucially involved as Indian people and communities grapple with problems with alcohol. PMID- 11414390 TI - "These young chaps think they are just men, too": redistributing masculinity in Kgatleng bars. AB - In the 19th century the BaKgatla polity was a chiefdom with a redistributional economy based on mixed agriculture. Sorghum beer was symbolic not only of the patrilineal core of their descent system and of the ideologies of reciprocity and redistribution, but also of masculinity and patriarchal control. With the establishment of a market economy, an industrial brewery and individual access to income, both beer and the act of drinking have been symbolically reconstructed. The ideology of redistribution was well suited to the support of the BaKgatla gerontocracy via alcohol production and consumption. The limits on production and consumption of beer inherent in the agricultural cycle and the control of young men's access by elders made alcohol an effective symbol of managerial competence from the limited context of household authority to that of the chiefdom as a whole. Today, young men's greater control of cash income has given them access to beer beyond the control of elders. As a result, the contrasting ideology of market exchange and competitive distribution of beer has contributed to the degradation of the power of seniors. After reviewing the historical background, this paper explores those changes. It argues that while the observed infrastructural changes have had a predictable impact on drinking behaviors and the symbolic structure of "seniority/masculinity", constructions of the "masculine community" in BaKgatla bars demonstrate continuity in key areas of mens' identities. If as anthropologists we see obvious discontinuities in behavior and ideology, the BaKgatla build selective bridges to "tradition" which seemingly ground the experience of change in relatively seamless continuity. PMID- 11414391 TI - "Take my water": liberation through prohibition in San Pedro Chenalho, Chiapas, Mexico. AB - This article explores changing views of alcohol in San Pedro Chenalho, a township in highland Chiapas, Mexico that has been profoundly transformed by the Zapatista democracy movement. The first part of the article provides an overview of drinking in Chenalho in the 1970s and 1980s when alcohol was still a strong symbol of community solidarity and an important part of healing ceremonies. The second part describes the period since the Zapatista uprising, a period in which indigenous women have begun to intensify their involvement in the political affairs of their communities. In their search for autonomy and to recover pride and dignity, both women and men supporters of the Zapatista movement reject alcohol as a symbol of political and economic domination. A commentary examines the relationship between the critiques of alcohol that have developed in Chenalho since the 1970s and political economy perspectives in alcohol studies. Both explanatory frameworks focus on the power structures in which alcohol sales and use are embedded. PMID- 11414392 TI - The relationship between within-interview contradictions and eliciting interviewer utterances. AB - OBJECTIVE: To determine whether interview practices associated with inaccurate reporting in laboratory analog contexts were also associated with inaccurate information in actual forensic contexts. METHOD: The forensic interview of a 5 year-old girl, an alleged victim of sexual abuse, was analyzed to explore interview practices associated with the retrieval of contradictory information. Content analyses of the child's responses focused on: (1) new informative details about the reported incidents; (2) contradictory details; (3) "central" and "peripheral" details; and (4) the types of utterances used to elicit each detail. RESULTS: The results illustrate how risky option-posing and suggestive utterances can be, as most (90%) contradicting details were elicited using option-posing and suggestive utterances and almost all (98%) of the contradicted and contradicting details were central, containing crucial information concerning the investigated allegation. No contradictory details were elicited in response to open-ended invitations. CONCLUSION: The findings demonstrate that poor interviewing practices can be associated with high levels of internal contradiction and should be avoided by forensic interviewers. To avoid contaminating children's reports and increase the likely accuracy of the information retrieved, moreover, interviewers should elicit as much information as possible using open-ended utterances, which tap free-recall memory. PMID- 11414393 TI - Association between early sexual abuse and adult HIV-risky sexual behaviors among community-recruited women. AB - OBJECTIVE: The first purpose was to determine whether sexual abuse involving penetration that occurred in childhood only, adolescence only, or both childhood and adolescence differently impacted whether community-recruited women had ever traded sex for money or drugs, their number of recent sex partners, and the number of times they had engaged in recent unprotected sex. The second purpose was to assess the mediating effects of adulthood rape, recent drug use, and recent sex with an injection drug user on these three HIV-risky sexual behaviors. METHOD: Women (n = 1,490) recruited from three US sites were questioned about their childhood and/or adolescent sexual abuse histories, adulthood rape experiences, recent drug use, and adult HIV-risky sexual behaviors via structured interviews. RESULTS: One-third of the women reported having experienced sexual abuse involving penetration in childhood and/or adolescence. Overall, regression analyses indicated a significant relationship between early sexual abuse and adult risky behaviors; rape in adulthood mediated this relationship for all three HIV-risky behaviors. Abuse that occurred in childhood only and abuse that occurred in both childhood and adolescence had a stronger impact on later risky behaviors than did abuse that occurred in adolescence only. CONCLUSIONS: Because childhood constitutes a critical period in individuals' sexual, social, and personal development, sexual abuse precipitated during this time may distort women's constructions of sex and sexuality. Women abused in childhood may therefore engage in HIV-risky sexual behaviors to a greater extent than women abused in adolescence as a result of these disruptions to their development. Rape in adulthood appears to intensify the effects of early sexual abuse, PMID- 11414394 TI - Guilt, isolation and hopelessness among female survivors of childhood sexual abuse: effectiveness of group work intervention. AB - OBJECTIVE: This study explores the effects of group work intervention on female survivors' senses of guiltlessness, affiliation and hopefulness. METHOD: Secondary comparative analyses of a large quasi-experiment-based clinical data base were accomplished (Richter, Snider, & Gorey): group work intervention (N = 78) and a waiting-list condition (N = 80). RESULTS: Group work was found to have beneficial effects on adult female survivors' appropriate sense of guiltlessness for their childhood sexual abuse, as well as on their sense of affiliation and hopefulness. Consistent across the three outcome measures of guilt/guiltlessness, isolation/affiliation and hopelessness/hopefulness, 16 to 18 of every 20 such women who participated in group work did better than the average woman in the waiting-list comparison group. Moreover, these apparent clinical benefits were maintained for 6 months (all p < .01). CONCLUSION: Such effects may be characterized as very large, and are generally larger than those previously observed in this field of practice that have typically been based on more general measures of depression, self-esteem or global symptoms. PMID- 11414395 TI - Clinical features of survivors of sexual abuse with major depression. AB - OBJECTIVE: This study examined differences in rates of trauma-related disorders between patients with histories of childhood sexual abuse and those without such histories in a sample of depressed outpatients. Another aim of this study was to determine whether childhood sexual abuse is associated with recent suicidal attempts, affect dysregulation and duration of index depressive episode, independent of posttraumatic stress disorder and borderline personality disorder. METHOD: Subjects were 235 treatment-seeking outpatients with major depression. Structured interviews were administered to assess for Axis I and Axis II disorders, childhood sexual abuse, and various clinical features. RESULTS: Patients with sexual abuse compared to those without sexual abuse histories had higher rates of comorbidity, primarily borderline personality disorder, posttraumatic stress disorder and multiple Axis I diagnoses. Childhood sexual abuse status was linked to a longer duration of the index depressive episode, independent of borderline personality disorder and/or posttraumatic stress disorder. However, childhood sexual abuse status was not independently related to affect dysregulation and suicidal attempts. CONCLUSION: The findings suggest that patients with histories of sexual abuse represent a subgroup of depressed patients who are at especially high risk for psychiatric morbidity and a prolonged episode of depression. PMID- 11414396 TI - Predicting the child-rearing practices of mothers sexually abused in childhood. AB - OBJECTIVE: Although empirical investigations have established a relationship between childhood sexual abuse and numerous long-term consequences, surprisingly little research has addressed the possible effects of childhood victimization on the later child-rearing practices of adult survivors. The present study examined hypothesized predictors of three parenting styles among adult survivors of sexual abuse as compared with adult children of alcoholic parents. METHOD: Forty-five clinical outpatients completed a questionnaire battery assessing experiences of childhood abuse, current economic and social resources, and parenting attitudes and practices. The child-rearing practices of participants were compared with those reported by a community sample of 717 mothers. Additional analyses examined the extent to which sexual abuse and its adult sequelae predicted the parenting behaviors reported by the present sample. RESULTS: Both sexual abuse survivors and children of alcoholics reported significantly higher rates of permissive parenting practices than mothers in the community sample. Multiple regression analyses further revealed unique relationships between sexual abuse and parenting, over and above the variance explained by physical abuse, current socioeconomic status, and the experience of growing up in an alcoholic home. Mothers' sexual abuse severity, social support satisfaction, and dysfunctional parenting attitudes moderated several of these relationships. CONCLUSIONS: The present findings suggest that sexual abuse and its adult sequelae may have negative consequences for the parenting practices of survivors, particularly for survivors' ability to provide their children with appropriate structure, consistent discipline, and clear behavioral expectations. Implications for the psychosocial development of survivors' children are discussed. PMID- 11414397 TI - The relationship between attitudes toward corporal punishment and the perception and reporting of child maltreatment. AB - OBJECTIVE: This study examined the effect of workers' attitudes toward corporal punishment on the workers' perception and reporting of child maltreatment. Three hundred twenty-five (325) potential entry level workers participated in this study. METHOD: Responding to vignettes in multi-item scales, study participants 1) rated their approval of parental discipline involving corporal punishment; 2) rated the seriousness of incidents of probable maltreatment; and 3) indicated whether or not they would report the incidents of maltreatment to child protective services. Data were analyzed using correlation analyses and multiple regression procedures. RESULTS: 1) Respondents with higher scores for approval of corporal punishment were less likely to perceive maltreatment; 2) respondents with higher scores for approval of corporal punishment were less likely to report maltreatment; 3) the likelihood that a respondent would report maltreatment was a joint function of the respondent's perception of the seriousness of an incident and approval of corporal punishment. CONCLUSION: Attitudes about corporal punishment are important predictors of reporting behavior. Social service agencies face a challenge to provide workers with training which will enable them to detect and report maltreatment despite workers' individual beliefs about discipline and punishment. PMID- 11414398 TI - Psychometric evaluation of the children's impact of traumatic events scale revised. AB - OBJECTIVE: To measure the internal consistency and construct validity of the Children's Impact of Traumatic Events Scale-Revised (CITES-R). METHOD: 158 sexually abused children, ages 7-12, and a nonabusive parent or primary caretaker completed a series of measures, including the CITES-R. A subsample of 17 repeated the assessment after 1 to 2 months with intervening treatment. RESULTS: Internal consistency for the CITES-R scales averaged .69. Associations with other measures were in predicted directions, although most were modest in size. Parent-report measures were virtually unrelated to any child-report measure, including the CITES-R. Change over time during treatment was noted on most symptom scales. CONCLUSIONS: The psychometric adequacy of the CITES-R was moderately supported. The main symptom scales of the instrument performed well. The performance of other scales was more variable. PMID- 11414400 TI - Sustainability. PMID- 11414399 TI - Relationships among parental beliefs in corporal punishment, reported stress, and physical child abuse potential. AB - OBJECTIVE: The present study examined the extent to which parental belief in the value of corporal punishment moderates the association between level of parenting stress and physical child abuse potential. Based on existing theory, it was expected that levels of parenting stress would be positively associated with physical child abuse potential among parents who reported high levels of belief in the value of corporal punishment. METHOD: Forty-one parents (25 general population and 16 at-risk parents) were assessed for belief in the value of corporal punishment, level of parenting stress, and physical child abuse potential using self-report measures. After removal of respondents due to response distortion or missing data, the final sample consisted of 31 parents with valid and complete protocols. Based on their responses on the study measures, respondents were categorized as either high or low on belief in corporal punishment and parenting stress. RESULTS: Level of parenting stress was positively associated with physical child abuse potential. As expected, the interaction of parenting stress and belief in the value of corporal punishment was significant. Level of parenting stress was positively associated with physical child abuse potential among parents who reported high levels of belief in the value of corporal punishment. In contrast, level of parenting stress was not associated with physical child abuse potential among parents who reported low belief in the value of corporal punishment. CONCLUSIONS: The present findings are consistent social information processing and stress and coping models of the etiology of physical child abuse, and underscore the importance of considering both parental cognitions and levels of parenting stress in assessing potential for physical child abuse. PMID- 11414401 TI - Humoral immune response following hepatitis B vaccine booster dose in children with and without prior immunization. AB - One hundred and twenty-three children who had received no, incomplete and complete primary hepatitis B vaccination but had negative or very low anti-HBs titer were immunized with a single dose of recombinant hepatitis B vaccine. Blood tests for anti-HBs were obtained at 30 +/- 5 days after the booster immunization. Twelve of 18 (66.7%) children without prior immunization (group 1) seroconverted following the single dose Seroconversion rates in children who had undetectable anti-HBs with incomplete and complete primary immunization (group 2 and 3) were 83.34% and 94.5%, respectively. All children with complete 3- dose vaccination but who had low anti-HBs titer (group 4) also seroconverted. This study confirmed that immunological memory, allowing a protective anamnestic response, lasted at least 8 years in children who had received primary HB immunization with undetectable anti-HBs. Therefore, we conclude that the booster dose after complete vaccination is not necessary in healthy children. PMID- 11414402 TI - Core promoter and precore mutants of hepatitis B virus: prevalence and clinical relevance in chronic hepatitis patients. AB - The present study was conducted to determine prevalence and exact type, as well as nucleotide position of the precore/core mutations of hepatitis B virus found in Thai patients diagnosed with chronic hepatitis and/or cirrhosis in relation to the clinical parameters established with the respective patients. To that end, 24 HBeAg-positive and 56 HBeAg-negative individuals were selected at random from a cohort of altogether 256 chronic liver disease patients. DNA was extracted from their blood sera, amplified by polymerase chain reaction using semi-nested primers and subjected to direct sequencing. Clinically, the HBeAg-positive chronic hepatitis patients displayed significantly higher transaminase levels than those negative for HBeAg. Our results showed 2 of the 7 (28.6%) PCR-positive HBeAg-positive sera displaying double mutations in the core promoter region at position 1762/64. The nucleotide sequences obtained from the 24 PCR-positive HBeAg-negative sera revealed 18 (75%) mutations in the core promoter region (1762/64), and/or 7 (29.2%) mutations at position 1753, and/or 6 (25%) mutations of the start codon (1814), and/or 8 of (33.3%) nucleotide 1896 turning codon 28 into a stop codon and one sample (4.2%) displaying a deletion between nucleotides 1758-1772. It is suggested that the mutations observed have an impact on the DNA secondary structure in such a way that successful transcription of the HBeAg gene is rendered impossible. To what extent this mutation influences the severity of chronic liver disease remains to be elucidated. PMID- 11414403 TI - Assessment of dengue hemorrhagic fever in Myanmar. AB - This study assesses the burden of dengue hemorrhagic fever (DHF) and effectiveness of an intervention package in Myanmar. Disability adjusted life years (DALYs) lost for fatal and non-fatal DHF cases and DALYs averted due to intervention for DHF from 1970-1997 were estimated. The data are based on reported cases and deaths from DHF extracted from annual reports of the Myanmar vector borne disease control program. Sensitivity analyses were performed for robustness of conclusions. DALYs lost from both fatal and non-fatal DHF cases in Myanmar were estimated as 83.83 DALYs per year per million population (range = 83.33-86.32) for the period 1970-1997. DALYs averted from DHF due to intervention were estimated as 134 DALYs per year per million population (range = 47-159). The burden of DHF in Myanmar for the selected year 1990 was 91.3 DALYs per year per million population (range = 90.1-96.5). A comparison was made with China, India and other Asian countries based on information provided by a World Bank study. PMID- 11414404 TI - Morbitz type I second degree AV block during recovery from dengue hemorrhagic fever. AB - Two patients with serologically-proven dengue virus infection and Morbitz type I second degree atrioventricular (AV) block are described. A 7 years old boy (patient 1) with grade 2 and a 7 years old girl (patient 2) with grade 3 illness were admitted to the hospital on the 3rd and the 5th day of the illness, respectively. Both had typical resentation for dengue hemorrhagic fever including fever, hepatomegaly, thrombocytopenia and signs of extravascular leakage. The 7 year old girl also had epistaxis and anemia (Hct 24%). Morbitz type I second degree and 2:1 AV block developed on day 7 (patient 1) and day 8 (patient 2) of the illness, both during recovery periods. Patient 1 also had occasional monomorphic premature ventricular contraction (PVC). There was no other abnormality in the 12-lead EKGS and echocardiograms showed normal ventricular systolic function in both. Other than mild hypokalemia (3.3 and 3.4 mgq/l), serum electrolytes were normal. Neither patients had elevation of serum creatine phosphokinase (CPK). In patient 1, exercise (on day 10) normalized AV conduction and abolished the PVC. Follow up EKG and physical examination at 10 months after the illness was normal. The rhythm in patient 2 resolved to 1st degree AV block (with occasional morbitz type I second degree at night) on day 12. In this patient, exercise resulted in shortening of the PR interval and Valsalva maneuver resulted in further PR prolongation. The patient was well at 1-month follow up with a mormal EKG. Morbitz type I second degree AV block during recovery from dengue hemorrhagic fever may be a transient functional impairment of the AV node, in which altered autonomic tone may play a role. PMID- 11414405 TI - Seroepidemiology of varicella-zoster in Pakistan. AB - The availability of safe and effective vaccines has renewed interest in the epidemiology of varicella worldwide. To date published data on the epidemiology of varicella in Pakistan is very scarce. Therefore, we conducted a study to determine the age-specific seroprevalence rate of varicella-zoster virus (VZV) antibodies in Pakistan. Between December 1997 and March 1998, 1,509 healthy volunteers aged between 1 month and 30 years were recruited from the Islamabad, Karachi, Lahore and Rawalpindi areas. Demographic information, socioeconomic status and past medical history were obtained by questionnaire. Serum samples were assayed for IgG antibodies against VZV by enzyme-linked immunosorbent assay. Overall 41.8% (600/1,435) of those tested were found to be seropositive for VZV antibodies. No difference was found in results obtained from the different cities. A higher seroprevalence was observed among women (45.2%) compared to men (39.6%). Seroprevalence rates increased with age and were 28.4% in those aged 0-5 years, 41.5% in the 6-10 year age group, 42.5% in the 11-15 year age group, 46.7% in the 16-20 year age group and 53.6% in those aged 21-30 years. Socioeconomic status was not a significant risk factor for VZV seropositivity. This is the first report of the seroepidemiology of VZV in Pakistan. The results indicate that seroprevalence of VZV increases with age in the Pakistani population studied. As in other tropical countries, there is greater susceptibility to varicella among the adolescent and young adult population. The results of this study suggest that these at-risk groups should be included in vaccination programs aimed at reducing the public health impact of varicella. PMID- 11414406 TI - A review of Hib epidemiology in Asia. AB - Meningitis due to an invasive Haemophilus influenzae type b (Hib) infection, has been previously perceived to be relatively uncommon in Asia. However, the incidence of disease and its impact may have been underestimated. In addition to a lack of microbiological facilities in some hospitals, difficulties in culturing the organism and the widespread use of antibiotics may have hidden the true incidence of the disease in some countries. Furthermore, the reported disease burden probably underestimates the incidence of Hib pneumonia. The epidemiology of invasive Hib disease for various Asian nations is reviewed in this paper. Hospital-based studies show that Hib is a major cause of bacterial meningitis and/or pneumonia in the Philippines, India, Thailand, Malaysia, Indonesia and Vietnam. Singapore and Hong Kong have a low incidence of infection compared with Western and other Asian nations. This low incidence is not due to a higher level of natural protective antibodies, but may be related to an interaction between environmental and genetic factors. Therefore the widespread belief that Hib infection is unimportant in Asia does not refer to Asia as a whole and possibly to Chinese patients only, and failure to recognize this has serious implications. The inclusion of Hib vaccine in the routine infant immunization schedule in many industrialized nations has significantly reduced the incidence of invasive disease. Recent studies have shown Hib vaccination is also effective in preventing invasive disease in children in developing countries. While population based data may be required to confirm the need for public-funded infant Hib immunization in Asia, its introduction in countries with a high incidence of Hib meningitis and/or pneumonia has the potential to significantly improve pediatric health and survival. PMID- 11414407 TI - Immune responses to measles immunization and the impacts on HIV-infected children. AB - This prospective cohort study was conducted to determine the seroconversion rate and the pattern of antibody response to measles vaccine administered at age 9 months in HIV infected and non-infected children born to HIV-1 seropositive mothers. Thirty children born to HIV-1 seropositive mothers and 3 born to HIV-1 seronegative mothers were recruited. One single dose of Schwarz strain of measles virus vaccine (Rouvax) was given to every child at 9 months of age. Clinical status and measles antibody levels were evaluated at the time just before vaccination, 2 and 12 weeks post-vaccination. Antibody was measured by an enzyme immunoassay commercial kit (Enzygnost, Dade Behring Manufacturer, Germany). Children were classified into 3 groups, groups 1 and 2 were children with and without HIV infection respectively. Group 3 children were those born to HIV-1 seronegative mothers. Of the 33 enrolled children, 16, 14 and 3 were classified as groups 1, 2 and 3 respectively. Four children, 2 of each, in groups 1 and 3 did not complete the study. Group 3 was excluded due to the small number of children recruited. There was no short term complication and no measles infection noted during the course of study. None of the children had pre-existing antibodies. The median (range) of CD4 count and CD4/CD8 ratio measured at the time of vaccination were statistically different between groups 1 and 2 children. Group 2 children had better antibody response than group 1 in terms of seroconversion rate and median of antibody levels at 12 weeks post-vaccination. Only 7 of 29 children (24.1%) had detectable measles antibodies at 2 weeks post vaccination. A decrease in antibody was noted in 2 symptomatic HIV infected children as their disease had progressed. Various potential predictors of measles vaccine responses in HIV infected children including CD4 count and CD4/CD8 ratio were not statistically different between the responders and non-responders. All 4 asymptomatic HIV infected children were responders. This study demonstrated that all of the children had already lost their maternal acquired antibodies at age 9 months. HIV infected children had a poorerantibody response to measles vaccine than the non-infected children. PMID- 11414408 TI - Opportunistic infections in the liver of HIV-infected patients in Thailand: a necropsy study. AB - Liver necropsy from patients infected with human immunodeficiency virus was analyzed in 117 cases. Wide ranges of opportunistic infections were recorded in 47%. Cryptococcosis (21.4%) was the most outstanding infection, followed by tuberculosis (16.2%), cytomegalovirus (5.1%) and penicillosis (3.4%). Non specific alterations of the liver tissues included fatty steatosis (49.6%), fibrosis (55.6%), portal inflammation and reactive hepatitis. Cases of chronic active and chronic passive hepatitis and one case of hepatocellular carcinoma were reported. In the infected liver, predominant pathological changes included granuloma and spotty necrosis, which were attributed to tuberculous hepatitis. Infection with Cryptococcus usually showed no associated pathological change. The sensitivity for the clinical diagnosis of Cryptococcus was 88.8% and specificity was 91.7%. For tuberculosis, sensitivity was 20% and specificity was 67.9%. PMID- 11414409 TI - Detection and molecular characterization of Vibrio vulnificus from coastal waters of Malaysia. AB - A total of 57 Vibrio vulnificus isolates from coastal water were characterized for their antimicrobial resistance, plasmid profiles and were typed by the PCR based techniques: a random amplification of polymorphic DNA (RAPD) method and the enterobacterial repetitive intergenic consensus sequence (ERIC) method. All isolates were susceptible to chloramphenicol, nalidixic acid, tetracycline and trimethoprim-sulfamethoxazole. Fifty-one isolates were resistant to one or more of the other antibiotics tested. Plasmid analysis indicated that only 18 isolates carried small plasmids of 1.6 to 16 megadaltons. Analysis of the RAPD and ERIC DNA fingerprints of the V. vulnificus isolates with Gel Compare and cluster analysis software revealed significant genetic heterogeneity among these isolates. The combination of RAPD and ERIC analysis allowed us to distinguish all isolates. Thus, the combination of the two techniques is recommended for epidemiological investigation. PMID- 11414410 TI - Development of ARI case management at primary and secondary level in southern Vietnam. AB - In southern Vietnam it is not uncommon that children under 5 years of age die from pneumonia. Reduction of severity and mortality has to rely on proper case management by mothers and health workers on both grass root level and referral level. The responsibility of training of clinical skills of ARI case management in the southern provinces of Vietnam has been delegated to Pediatric Hospital N1 (PHN1) Ho Chi Minh City (HCMC) by Ministry of Health. A pilot project was carried out by the Danish-Vietnamese Study Group. The immediate objects were: to provide basic epidemiological information about ARI in southern Vietnam, to develop training modules and case management intervention modules at primary and secondary level in order to enable mothers, village workers, health post staff and district hospital emergency department staff to treat moderate and severe pneumonia and acute bronchitis in accordance with the WHO management guide for ARI and to evaluate the effect of those modules after implementation in a limited number of communes. The modules were developed at PHN1. Ten commune health stations were carefully selected. The purpose of the project and the conditions for taking part had been explained to the health workers. The doctors and other commune health workers from the 10 commune health stations and doctors from the connected district hospitals attended the training courses at PHN1, HCMC and also at the belonging provincial hospitals. Essential equipment was provided and a pharmacy with essential drugs established. The registered health statistics was collected yearly during on site visits. The local doctors and commune health workers gave seminars for mothers in the villages of the 10 project communes. The mothers' knowledge, attitude and practice (KAP) was tested in interviews before and two months after the seminars had taken place. The spread of KAP was measured by random interviews of mothers six month later. In the interviews information on social conditions was obtained. The mothers' KAP had risen by 25% two months after attending the seminars. A further increase of KAP by 5-10% within the untrained group appeared in a survey 4-6 months later. It was not possible to obtain reliable statistics on morbidity or mortality of ARI in the project area. PMID- 11414411 TI - Risk factors for penicillin-resistant Streptococcus pneumoniae acquisition in patients in Bangkok. AB - To identify risk factors for acquisition of penicillin-resistant Streptococcus pneumoniae (PRSP) in patients in Bangkok, using a case-control study, the study included patients with clinical specimens which grew S. pneumoniae during January to December 1997, treated at a teaching hospital in Bangkok. Penicillin susceptibility was determined by E-test and strains with MIC of > 0.1 microg/ml were considered resistant. Cases were the patients who had PRSP, and patients who had penicillin-susceptible S. pneumoniae (PSSP) were controls. The study variables included age 15 years or younger, immunocompromised status, ventilatory support, and antibiotic use or hospitalization within the previous 3 months. There were 73 cases and 51 controls. Their ages were 0 to 87 years, with median age of cases 4 and controls 49 years. Pneumonia was the most common type of infection, being 47% in cases and 45% in controls. Univariate analysis revealed significant association of PRSP acquisition with previous antibiotic use (p<0.0001), age < or = 15 years (p=0.001) and previous hospitalization (p=0.002). Logistic regression analysis in order to adjust for confounding effects showed that the only significant risk factor was previous antibiotic use (OR 18.4; 95% CI 6.2-54.6). The major risk factor for acquisition of PRSP in this study population is recent antibiotic use. Decreased antibiotic use would reduce risk of acquisition of PRSP. PMID- 11414412 TI - Detection of Mycoplasma pneumoniae from cerebrospinal fluid samples of pediatric patients. AB - Central nervous system manifestations are probably the most frequent extrapulmonary complications of infections due to Mycoplasma pneumoniae, occur mostly in children. In this study, we attempted to isolate M. pneumoniae and to detect the organism by polymerase chain reaction (PCR) from cerebrospinal fluid samples (CSF) of pediatric patients. Of the 244 CSF samples, no M. pneumoniae was isolated. Six (2.5%) of the CSF samples were positive by PCR amplification. More effort are necessary to isolate the organism from CSF samples in order to ascertain the role of M. pneumoniae in causing neurological complications. PMID- 11414413 TI - Isolation and polymerase chain reaction detection of Mycoplasma pneumoniae in Malaysian patients with respiratory tract infections. AB - Isolation and polymerase chain reaction (PCR) were performed for detection of Mycoplasma pneumoniae from respiratory tract specimens obtained from 200 adult and 200 pediatric patients. M. pneumoniae was isolated from bronchoalveolar lavage fluid of 1(0.5%) adult patient and 4(2.0%) tracheal aspirates of pediatric patients. PCR was positive for only one (0.5%) broncoalveolar lavage fluid of an adult patient and fifteen (7.5%) tracheal aspirates of pediatric patients. This study suggested that M. pneumoniae was more frequently detected in pediatric patients and PCR appears to have advantages over isolation, in terms of rapidity and sensitivity. PMID- 11414414 TI - Enzyme biotypes of Helicobacter pylori isolated from Penang, Peninsular Malaysia. AB - A total of 52 clinical strains of Helicobacter pylori were characterized on the basis of preformed enzyme production with API ZYM kits. Using the biotyping schemes as defined by Reina and Alomar (1989), Kung et al (1989) and Matsumoto et al (1996), 15.3% (8/52), 13.5% (7/52) and 11.5% (6/52) of the isolates were not biotypable, respectively. Two enzymes, valine arylamidase and cystine arylamidase could be additionally used to differentiate between isolates. Our isolates were either negative or positive for both the enzymes or positive only for cystine arylamidase. We propose the incorporation of these two enzymes into the Matsumoto et al (1996) biotyping scheme to biotype strains into additional enzyme biotypes. PMID- 11414415 TI - Application of ribosomal RNA gene restriction patterns analysis and pulsed-field gel electrophoresis in distinguishing Salmonella weltevreden isolates in Malaysia. AB - A representative sample of 20 isolates of Salmonella weltevreden strains from stool cultures of patients admitted at the University Hospital, Kuala Lumpur, Malaysia were analyzed. All the strains were susceptible to ampicillin, ceftriaxone, ciprofloxacin, chloramphenicol, tetracycline, trimethoprim, gentamicin and co-trimoxazole. Ribosomal RNA gene restriction pattern analysis of PstI-digested DNA gave three ribotypes while pulsed-field gel electrophoresis (PFGE) analysis of XbaI-digested DNA gave ten distinct profiles. PFGE was more discriminative than ribotyping in distinguishing the strains. The majority of the strains analyzed were very closely related with similarity coefficient values ranging from 0.8 to 1.0. Both PFGE and ribotyping could distinguish one of the strains which was obtained from a patient following a bone marrow transplant for beta-thalassemia major, indicating that this particular strain was unrelated to the rest of the strains from patients with acute gastroenteritis. PMID- 11414416 TI - The diagnostic value of the ELISA-Ty test for the detection of typhoid fever in children. AB - A study on the reliability of an enzyme linked immunosorbent assay (ELISA) for the detection of typhoid fever, the ELISA-Ty test, was conducted, comprising 44 children suffering from bacteriologically confirmed typhoid fever based on the finding of a positive blood culture for Salmonella typhi, 44 children with fever caused by diseases other than typhoid fever based on the finding of negative culture of blood, urine and stool for S. typhi, and 120 healthy children as controls. This ELISA-Ty test measures the concentration of IgM and of IgG against S. typhi in serum. This test is an indirect ELISA test, based on a method that makes use of a mixture of OMPs (outer membrane proteins) in equal proportion serving as antigen, obtained from different strains of S. typhi which are prevalent locally, peroxidase goat antihuman IgG or IgM (Sigma) as conjugate and orthophenylenediamine (Sigma) as chromogen of the substrate. The result of the test was obtained through the assessment of the end product, using a micro ELISA reader (Behring) at wave length of 490 nm. The data revealed that the mean absorbent values found in children with typhoid fever, for IgM and IgG, were significantly higher (p < 0.05) when compared to those in children with non typhoid fever as well as to those found in children of the control group. The results of this study confirm that the ELISA-Ty test has a high reliability for the detection of typhoid fever in children, based on the finding of a degree of diagnostic sensitivity as high as 95.45% and 90.91% for respectively IgM and IgG, a diagnostic specificity as high as 93.33% for IgM as well as for IgG, a high diagnostic efficiency (94.32% for IgM and 92.05% for IgG), a high diagnostic positive predictive value of 93.33% for IgM and 93.02% for IgG, high negative predictive values of 95.35% for IgM and 91.11% for IgG for use under clinical as well as under field conditions. PMID- 11414417 TI - Epidemiology of invasive meningococcal disease in 13 government hospitals in Thailand, 1994-1999. AB - This study was conducted to elucidate the magnitude of problem and the clinical course of invasive meningococcal infection from 13 government hospitals in Thailand between 1994 and 1999. Thirty-six strains of Neisseria meningitidis were isolated from 16 blood and 24 cerebrospinal fluid specimens; 4 patients had positive culture in both blood and CSF. Of the 16 strains, 9 (56.3%) were serogroup B. Seventy-one and eighty-four percent of the isolates were susceptible to penicillin and cefotaxime/ceftriaxone respectively. Five out of six penicillin nonsusceptible strains were found to be relatively resistant to penicillin with the MIC of 0.125 microg/ml. Of 33 patients whose medical records were available, 21 were males and 12 were females, with a mean age of 11.2 years. Fifteen patients (45.5%) presented with meningococcemia and 18 patients (54.5%) presented with meningococcal meningitis. Hypotension and purpura were found in 24.2% and 33.3% of patients respectively. The overall mortality rate was 9.1%. In conclusion, meningococcal disease is not common in Thailand, meningococcemia is a life-threatening condition whereas meningococcal meningitis is much less severe. The prevalence of meningococci relatively resistant to penicillin seems to be increasing. PMID- 11414418 TI - House dust mite fauna in the Klang Valley, Malaysia. AB - Allergy to house dust mites (HDM) is an important cause of asthma and rhinitis in Malaysia. This study was carried out to evaluate the dust mite fauna in the Klang Valley. Dust samples were collected from 20 houses from March 1994 to February 1995. Thirty-three dust samples from mattresses were examined monthly for the occurrence of HDM. A total of 22 species in 9 families of HDM was identified. The most common and densely populated species was Blomia tropicalis with an average density of 8,934 mites/g of dust. Dermatophagoides pteronyssinus was the next in abundance, followed by Malayoglyphus intermedius. All houses surveyed were found to be infested with HDM and every house had at least 6 species of HDM. B. tropicalis and D. pteronyssinus were found in all mattresses. HDM in the Klang Valley were found to be highly prevalent and present in high densities. In this study, counts of D. pteronyssinus was found to exceed the proposed exposure threshold of 500 mites/g dust, for triggering acute asthma. Although counts of B. tropicalis exceeded D. pteronyssinus, no conclusion could be made because there is currently no exposure threshold for triggering acute asthma, for this species. Monthly distribution of B. tropicalis and D. pteronyssinus showed 2 peaks and 4 peaks, respectively. The major peak for D. pteronysinus was in January 1995 whereas for B. tropicalis, the major peak was more variable and occurred between November 1994 to January 1995. Both the species showed minor peak in April 1994. PMID- 11414419 TI - First report on sensitization to allergens of a house dust mite, Suidasia pontifica (Acari: Saproglyphidae). AB - A species of house dust mite, Suidasia pontifica, was recently shown to produce allergens affecting man. The species may be as important as other allergen producing mite in sensitization and causing allergic symptoms in Malaysians. Surveys conducted demonstrated that 80% of the houses surveyed were positive for this mite with densities ranged from 2 to 50 mites per gram of dust. Colonies of the species has been successfully established and materials from those colonies have been used to produce extracts for studies on sensitization to the mites. A total of 85 suspected allergic rhinitis patients were tested and 74.1% demonstrated positive reactions. Extract of this mite should be considered for routine diagnostic testing and possible immunotherapy. PMID- 11414420 TI - Artemether, an effective new agent for chemoprophylaxis against shistosomiasis in China: its in vivo effect on the biochemical metabolism of the Asian schistosome. AB - Conventional drug chemotherapy against human schistosomiasis currently relies on treatment with praziquantel to eliminate adult schistosome worm pairs. The use of praziquantel for control purposes is limited, however, by high rates of post treatment re-infection with subsequent parasite egg deposition and host end-organ damage. Artemether, a methyl ether derivative of the anti-malarial drug quinghaosu, was discovered recently to also have anti-schistosomal properties. Because artemether selectively targets the larval migratory stages of the parasite, known as schistosomulae, it blocks the development of ovipositing adult schistosome worm pairs in the vasculature. On this basis, we have since shown in clinical trials conducted in China that artemether has proven benefit as an agent for chemoprophylaxis. In vivo studies using laboratory animals suggest that artemether causes damage to the tegument and musculature of schistosomulae. Artemether may exert its helminthotoxic effect through synergy with hemin or related heme-containing compounds. Schistosomes recovered from artemether treated laboratory animals have increased glycogen phosphorylase activity, but decreased glucose uptake. These findings may account for their decreased glycogen content, relative to schistosomes recovered from untreated laboratory animals. The artemether-damaged schistosomes also have decreased activities of a number of enzymes and enzyme systems, including glycolysis. This might suggest common pathways by which artemether may target human parasites that live in the bloodstream. PMID- 11414421 TI - Rickettsial infection in five remote Orang Ulu villages in upper Rejang River, Sarawak, Malaysia. AB - People in 5 Orang Ulu villages in Sarawak, Malaysia were tested for rickettsial infection by Weil-Felix reaction and by indirect immunoperoxidase reaction. Of those surveyed 9.6% were positive for typhus. Of the positives, 3.8% were positive for tick typhus (7/11), scrub typhus (4/11) or endemic typhus (1/11). The incidence of typhus was higher among semi-nomadic Penans compared with the settled Kayans. PMID- 11414422 TI - Epidemiology of human ancylostomiasis among rural villagers in Nanlin County (Zhongzhou Village), Anhui Province, China: II. Seroepidemiological studies of the age relationships of serum antibody levels and infection status. AB - Anti-hookworm antibody serologic responses were measured in residents of an Anhui provincial Chinese village where Ancylostoma duodenale is the predominant hookworm. Antibody responses were measured against either soluble infective third stage larval (L3) or adult antigens. Immunoglobulins of the IgG class, especially IgG4 correlated with both the prevalence and intensity of A. duodenale hookworm infections. In contrast, there was an inverse correlation with IgM, but no correlation with IgA or IgE. Circulating IgG4 antibody responses might serve as a surrogate marker for active A. duodenale hookworm infection. PMID- 11414423 TI - Prevalence of antibodies to Toxoplasma gondii among urban and rural residents in the Philippines. AB - A survey of antibodies to Toxoplasma gondii was carried out among residents in urban (Metro Manila) and rural (Mindoro and Leyte) areas in the Philippines. A total of 1,173 serum samples were examined for Toxoplasma antibody by an ELISA method. The overall seropositivity was 11.1% (n=904, 12.4% in males, 10.0% in females) in Metro Manila, 61.2% (n=152, 63.3% in males, 53.1% in females) in Mindoro, and 30.1% (n=113, 34.3% in males, 22.5% in females) in Leyte, indicating significantly higher (p<0.001) seropositivities in rural than urban settings. No significant differences in seropositivities were observed between males and females. In each group, seropositivity tended to increase with age of the subjects. PMID- 11414424 TI - Surface ultrastructure of excysted metacercariae of Haplorchis taichui (Trematoda: Heterophyidae). AB - The morphology of newly excysted juvenile Haplorchis taichui was studied using a light and a scanning electron microscope (SEM). The reproductive organs were well developed. The whole body surface was covered with numerous transverse rows of scale-like spines, which had 3-11 points at the tip. The spines on the dorsal were similar with those on the ventral surfaces in shape, size and number of points. The spines in anterior were digitated into 10-11 points, then 8-9 points and 7 points. The presence of 3 points in each spine was observed in the area adjacent to the excretory pore. Two types of sensory papillae existed throughout the body: type I, ciliated knob-like swellings and type II, round swellings of the tegument. The rapid maturation of H. taichui results from the development of both internal and external organs during the newly excysted stage. PMID- 11414425 TI - Effect of intestinal helminthiasis on nutritional status of schoolchildren. AB - This study was conducted in a rural agricultural area in Siniloan, Republic of the Philippines. The subjects were the school children. The nutritional status of 58 children infected with helminthiasis (Ascaris lumbricoides and Trichuris trichiura) was compared with that of 19 uninfected controls. Prevalence of Ascaris and Trichuris was 40.3% and 71.4% respectively, and 36.4% of infected children had both Ascaris and Trichuris infections. Statistically significant evidence of an adverse effect of helminthiasis on serum albumin levels was found, but no child had inadequate levels of other nutrients. Although helminthic infections increase the level of immunoglobulin E (IgE) in children endemically exposed to these parasites, there was no significant difference in the serum IgE among Ascaris or Trichuris infected groups in this study. PMID- 11414426 TI - Economic evaluation of water iodization program in Thailand. AB - In Thailand, iodine deficiency disorders (IDD) are endemic in 57 out of 75 provinces with an estimated 15 million people at risk of IDD. A three pronged control program with iodized salt, iodized water and iodized oil capsules is being implemented. The water iodization program is both school based and household based. In the household, the residents are given iodine solution, two drops of which is to be added to 10 l of drinking water. In the schools, in addition to this method, an iodinator is used. This releases a fixed amount of iodine into the drinking water. This study examines the cost of the water iodization program in Thailand for the year 1996 in terms of cost per beneficiary, cost per microg iodine consumed daily and cost per goiter person years averted. We used a discount rate of 5%. Field visit and interviews of health personnel from Ministry to village level were conducted to gather primary data. Review of existing papers and reports of the Department of Health, Government of Thailand was done for secondary data. The costs included the capital cost of equipments, initial training and the recurrent costs of potassium iodate, proportional salaries of personnel involved, monitoring and communication activities. The cost per beneficiary of school based iodinator method (US$ 0.72) and school based drop method (US$ 0.64) were similar and much higher than the household based approach (US$ 0.12). The cost per microg of iodine consumed daily was ten times higher in the school based approach (US$ 0.01) compared to the household approach (US$ 0.001). The cost per goiter case averted for the whole strategy of water iodization was US$ 194.50. Water iodization appears to be a low cost intervention. However, the need for behavioral modification raises the issue of long term sustainability. PMID- 11414427 TI - Follow up of water use in a tin mining area affected with arsenic poisoning. AB - Ron Phibun district in southern Thailand has been known as an endemic area for arsenic contamination. The government has been trying to improve the situation by encouraging the use of rainwater and piped water. This study aimed to document the change of water use and to identify factors associated with safe water use in 1997 compared to that in 1994. Home visits and face-to-face questionnaire interviews were undertaken. Information on water use for drinking, cooking, washing food and washing utensils in 1994 and 1997 was obtained. Among 3,849 households from which data could be obtained (estimated 79% of total households), the percentages of using safe water (including water from bottled rain water, piped and artesian well water) for drinking and cooking rose from 72.5 and 57.9 in 1994 to 93.6 and 80.9 in 1997, respectively. The percentages for washing foods and for washing utensils rose from 28.6 and 20.5 to 59.1 and 53.8, respectively. In 1997, percentage of households using piped water for drinking and cooking was still low (3.6 and 12.3) compared to those using piped water for washing food and utensils (39.1 and 43.6). Multivariate analysis shows that independent factors of the household predicting safe water use are: high arsenic area, near main road and having piped water installed. The influence of these factors (as judged by the level of odds ratio) operates more or less equally on water use for all purposes, except that installation of piped water has more influence on washing water than drinking and cooking water. We conclude that safe water supply in the area is still inadequate. Even if piped water is installed, it is often not used for drinking and cooking. The reasons for not using piped water for drinking and cooking need to be identified. PMID- 11414428 TI - The prevalence of periodontal disease and oral hygiene care in Savannakhet Province, Lao People's Democratic Republic. AB - The objective of this study was to evaluate the prevalence of periodontal diseases in Savannakhet Province, Lao PDR. This survey was conducted 14 years after the country reformed its economic structure towards the free market. Thirty villages were picked by simple random sampling from a total of 1,560 villages. Two thousand four hundred and fifty-three individuals, aged twelve and over, were interviewed about oral hygiene practice, and were examined for periodontal health status using the community periodontal index of treatment needs (CPITN) index. Forty percent of the sample had regular practices, 56.5% occasional practices and 3.5% were ignorant about oral hygiene. Oral hygiene practices were performed in the morning by using a toothbrush (61.1%) and the index finger (24.8%). The cleaning agent was water alone for 68.6% and salt powder for 28%. The prevalence of healthy gingiva was nearly zero in all age groups. Gingival bleeding was very low. The most common periodontal problem was calculus deposits, found in 3-5 sextants of all age groups. The pocket depth was 4-5 mm on average; however a depth of more than 6 mm was not observed. PMID- 11414429 TI - Validation of the Short Michigan Alcoholism Screening Test Thai version in northeastern Thailand. AB - This study aimed to validate the Short Michigan Alcoholism Screening Test-Thai version-12 items (SMAST-T) compared to the psychiatrist's diagnosis based on DSM III-R criteria, as a gold standard, among psychiatric patients residing in the northeast of Thailand. Sixty-one pairs of male cases with alcohol use disorders (AUD) and controls were collected through routine examination of psychiatric outpatients, 18-65 years old, who visited the Khon Kaen Psychiatric Hospital or the Northeast Drug Dependence Treatment Center, located in Khon Kaen Province, northeast Thailand, between November, 1996 and February, 1997. Controls were matched for each case in terms of age (+/-5 years), province and urbanization of residence. They were interviewed using the SMAST-T and a structured questionnaire. Receiver operating characteristic analysis revealed that the optimum cutoff point of 4 or over on the SMAST-T yielded the best sensitivity and specificity (96.7% and 95.1%, respectively), along with the area under the curve of 0.994, indicating that it was sensitive and specific in discriminating AUD from the non-AUD patients. Validation of the SMAST-T suggests its applicability as a screening interview questionnaire to detect AUD among northeast Thai males, with a specific cutoff point. PMID- 11414430 TI - Leptin concentration in relation to body mass index (BMI) and hematological measurements in Thai obese and overweight subjects. AB - The weight, height and body mass index (BMI), including waist/hip ratio, serum leptin and hematological parameters of 48 male and 166 female overweight (BMI > or = 25.00) Thai volunteers who came for a physical check-up at the Out-patient Department, General Practice Section, Rajvithi Hospital, Bangkok during the period March-October 1998, were investigated. There were statistically significantly higher levels of serum leptin, mean corpuscular mean corpuscular hemoglobin concentration (MCHC) and mean corpuscular volume (MCV) in the overweight than in the control subjects. The median serum leptin concentration in overweight subjects was 19.6 (2.0-60.0 ng/ml) compared with 9.0 (range 1.0-30.0 ng/ml) in the control subjects (p < 0.001). The medians of leptin in overweight and obese males were significantly higher than those of overweight and obese females. 66.7% (32 out of 48) of overweight and obese males were found to have elevated leptin levels, while 87.3% (145 out of 166) were found in overweight and obese females. Anemia was found in 18.7% of female overweight and obese subjects, using hemoglobin as an indicator. Significant associations were found between weight, height, BMI, waist, hip, waist/hip ratio, hemoglobin, hematocrit, and serum leptin in both male and female overweight subjects. A negative correlation was found between serum leptin and hemoglobin, and hematocrit in both overweight and obese subjects. PMID- 11414431 TI - Successful prophylaxis of intracranial hemorrhage in infants with severe congenital factor VII deficiency. AB - During the period 1984-1992, 2 severe cases (1 male, 1 female) of congenital F VII deficiency with intracranial hemorrhage (ICH) were referred to the Department of Pediatrics, Siriraj Hospital Bangkok, Thailand at the ages of 1 and 3 months old. They both responded very well to fresh frozen plasma (FFP) transfusion therapy. Subsequently, both had repeated episodes of ICH (repeated ICH) 5 and 6 times, despite the 10-14 days of replacement therapy for each episode and eventually died at the ages of 11 and 13 months. Since September 1996, another 2 severe cases (2 females) of congenital F VII deficiency who had ICH within their first month of life were referred to us. In order to prevent repeated ICH, we started a prophylactic regime after the second episode of ICH, by giving FFP 10 ml/kg twice a week. The average duration of follow up was 21 months (at 8 and 34 months). All of them (aged 14, and 38 months old) are doing well at this time and free from repeated ICH. From this observation, if there is FFP available, this regime is an effective way to prevent repeated ICH in infants with severe congenital Factor VII deficiency. PMID- 11414432 TI - A randomized clinical trial of combinations of artesunate and azithromycin for treatment of uncomplicated Plasmodium falciparum malaria in Thailand. AB - Recently, a combination of artesunate and mefloquine has proved effective, although is contraindicated in early pregnancy and young children. Azithromycin, a widely used antibiotic and has antimalarial effects, replace mefloquine as a new alternative antimalarial regimen. Two hundred and two uncomplicated falciparum malaria patients were randomly assigned to 1 of 3 regimens. Patients in group I (n = 68) received artesunate 200 mg once daily for 3 days, group II (n = 67) received artesunate 200 mg together with mefloquine 10 mg/kg on the first 2 days and artesunate 200 mg together with mefloquine 5 mg/kg on the third day, and group III (n = 67) received artesunate 200 mg together with azithromycin 50 mg once daily for 3 days. The 28 day cure rates were 44, 98 and 56%, respectively. The median time to recrudescence was significantly longer in group III. In conclusion, a combination of artesunate and azithromycin might be useful in treating children in whom bacterial and malarial infections may be concomitant. However, further work is required in order to enhance its clinical efficacy. PMID- 11414433 TI - Sequence diversity of serine repeat antigen gene exon II of Plasmodium falciparum in worldwide collected wild isolates. AB - Field isolates of Plasmodium falciparum collected from endemic areas of Southeast Asia, Solomon Islands, tropical African countries and Brazil were analyzed for the genetic diversity of the exon II of serine repeat antigen gene (SERA) by sequencing of genomic DNA. Of sixty-nine isolates, as compared to the reported FCR3, K1 and Honduras-1 types of exon II sequences, 5, 9 and 20 new allelic forms were found in 23 isolates of the FCR3 type, 36 of the K1 type and 10 of the Honduras-1 type. A group of novel non-synonymous substitutions, 4 new insertions and 3 new deletions of octamer units were found in the octamer repeat region (OR) of the exon II, and most of them clustered within a 40-residues domain. An octamer "SNPVSSEP" revealed in the OR was confirmed as a new repeat unit. Based on the sequences of the serine repeat region (SR) of the exon II, the allelic forms of the Honduras-1 type were conjectured to be the recombinant forms between the K1 type and FCR3 type. The allelic forms of K1 type with less or more repeat serine residues in the serine stretch of the SR than the reported 21 serine residues had most of the variations in the OR. Moreover, a biased geographical distribution of allelic forms was observed. Isolates from African and Southeast Asian countries accounted for most of the new allelic forms (29/33). All of the three types were detected in Southeast Asia but none of the FCR3 type in Africa. One of two groups of FCR3 new allelic forms was found solely in Brazil while another was mainly in Solomon Islands. PMID- 11414434 TI - Investigation of malaria prevalence at National Thermal Power Corporation, Shaktinagar, Sonbhadra District (Uttar Pradesh), India. AB - Malaria in industrial complexes is promoted by extensive mosquitogenic potential generated by excavations and importation of parasite through migratory labor. The National Thermal Power Corporation (NTPC), Shaktinagar, Sonbhadra district was surveyed for malariogenic conditions from 1994 to 1996. The major mosquito breeding sites were drains, storm-water drains, lakes, outside tanks, overhead tanks, sluice-valve chambers, ornamental tanks, wells, pit wells and water reservoirs, etc. Anopheles culicifacies was the major vector of malaria in this area. Sibling species identification of An. culicifacies revealed that species C predominated during the transmission season and responsible to transmit malaria. Insecticide susceptibility tests against An. culicifacies sl showed that An.culicifacies population was 100% susceptible to malathion, fenitrothorn and deltamethrin while it was found 44% resistant to DDT. The malaria cases recorded in 1994, 1995 and 1996 were 847, 590 and 409 respectively. In vitro study on P. falciparum cases showed that 41, 70, 50% of the isolates tested were resistant to chloroquine in 1994, 1995 and 1996 respectively while an in vivo follow-up study showed 20-30% P. falciparum cases resistant to chloroquine. An integrated approach involving alternate vector control measures along with judicious use of insecticides has been suggested to bring down malaria in industrial complexes. PMID- 11414435 TI - Is there any artemisinin resistance in falciparum malaria? AB - We reported two cases of complicated falciparum malaria who had poor response to artesunate with delayed parasite clearance times. They were splenectomized patients who were treated with high doses of artemisinin derivatives. Our cases showed the importance of the spleen in the clearance of malaria parasites and had different clinical outcome, one fatal and one recovery. The host factors, the parasitemia count, the quality of antimalarial chemotherapy and blood level of the antimalarial drugs must be considered in relation to the causes of the delayed clearance of parasitemia. PMID- 11414436 TI - Absence of knobs on parasitized red blood cells in a splenectomized patient in fatal falciparum malaria. AB - We present a case report of fatal falciparum malaria of a splenectomized adult Thai patient. The patient developed high peripheral parasitemia and showed signs of severe malaria with multiorgans involvement. Ultrastructure of Plasmodium falciparum-infected red blood cells in a fatal splenectomized patient and pathological features are reported for the first time with special emphasis on the role of the spleen as a modulating cytoadherence phenotype of parasitized red blood cells (PRBC). In this patient, adherence of the PRBC to the vascular endothelium of brain, kidney and lung including blood circulating cells, was noted, despite the absence of knob on the surface of the PRBC. PMID- 11414437 TI - Parasitological, pathological and functional studies on the effects of IFN alpha 2b in murine hepatic schistosomiasis. AB - The effects of different doses of Interferon alpha 2b (IFN alpha 2b), alone and in combination with praziquantel (PZQ), on hepatic schistosomiasis were tested. An experimental murine model of hepatic schistosomiasis was used. Four parameters were assessed; hepatic fibrosis by estimation of OH-proline content/g dry weight liver, hepatocyte proliferative activity by the PCNA/LI, schistosomal egg load by digesting parts of the liver by KOH and hepatocyte function by measuring parenchymal liver enzyme levels. IFN alpha 2b was found to increase hepatic fibrosis in a dose dependent manner both alone and in combination with PZQ. An augmentation of the regenerative activity of the liver was observed. A reduction in the number of the granulomas and egg counts was observed only when PZQ was added. However, no effect on the size of the granulomas was observed apart from the normal process of modulation. Caution should be exercised when treating patients with concomitant hepatic schistosomiasis and hepatitis with IFN alpha 2b as it increases both hepatocyte regenerative activity and hepatic fibrosis; two main components of cirrhosis. PMID- 11414438 TI - Study on bionomics of principal malaria vectors in Kratie province, Cambodia. AB - A cross-sectional study on bionomics of malaria was carried out in Kamlek village, Snuol rubber plantation, Kratie Province where the population had a high prevalence of malaria. Mosquitos were collected monthly between April to September 1998 from 18.00 hr to 6.00 hr by human bait and animal bait. Survey for larva breeding places was also performed. A total of 13 species of Anopheline mosquitos was collected, manly Anopheles philipinensis, An. aconitus, An. annularis, Ain. barbirostris, An. hyrcanus gr, An. jamesii, An. kawari, An. tesselatus, An. umbrosus and An. vagus. They were in various densities with the latter species being the most abundance. Among the vector species. An. maculatus was the most commonly collected with a density of 0.55 per man per night and peak biting was between 20.00 hr and 21.00 hr. The density of An. minimus and An. dirus were 0.11 and 0.74 per man per night with the peak biting time 24.00-01.00 hr and 20.00-21.00 hr respectively. Observation of host preference and feeding habit revealed An. maculatus to be exophagic, anthropophilic and 20 endophilic while An. minimus was both endophilic and exophagic and anthropophilic. An. dirus was endophilic and anthropophilic, these three species were not found positive for malaria infection. However, due to the limited time of survey and an exceptionally low rainfall in the rainy season, an inadequate sample was obtained for analysis. Therefore further investigation is required for at least a year for more information. PMID- 11414439 TI - Correlation of glutathione S-transferase and DDT dehydrochlorinase activities with DDT susceptibility in Anopheles and Culex mosquitos from northern Thailand. AB - Comparative DDT-susceptibility status as well as glutathione S-transferase activity and DDTase activity of Anopheles minimus (A). An. annularis and Culex quinquefasciatus were investigated to ascertain the role of these enzymes in DDT resistance. The standard WHO susceptibility test kits was used to discriminate between resistant and susceptible populations. GST activity was measured in microtiter plates whereas DDTase activity was determined by HPLC quantitation of DDT metabolites. This is the first report of DDT-resistance in the Thai malaria vector, An. minimus species A. A positive correlation of DDT-resistance and DDTase activity was observed in this species as well as in the suspected vector, An. annularis. However, GST activity was not correlated to DDT-resistance in either species. Statistical analysis and scatter plots demonstrated the non correlation between DDTase and GST activity in An. annularis. Studies in Culex quinquefisciatus revealed difference in GST/ DDTase and the relationship to DDT resistance compared to the anopheline species. The Culex GST activity is correlated to DDTase activity. These results suggested that a positive correlation of GST and DDTase activity might be species dependent. PMID- 11414440 TI - Highly conserved nucleotide sequence and its deduced amino acids of the 5' noncoding region and the capsid protein of a Bangkok isolate dengue-3 virus. AB - The dengue-3 virus genome encodes an uninterrupted open reading frame (ORF) flanked by 5' and 3' non-coding regions. The order of proteins encoded in dengue 3 virus ORF, as with other flaviviruses, is: Cap 5'-C-prM-E-NS1-NS2A-NS2B-NS3 NS4A-NS4B-NS5-3'. The nucleotide sequence of the 5'-noncoding region and the capsid protein of dengue-3virus (a Bangkok isolate: CH53489 isolated by USAMC AFRIMS in 1973) has been analyzed in both forward and reverse directions. The PCR based cycle sequencing technique by the enzymatic method of Sanger et al (1977) using a sequencing primer 5'-end labeled with gamma32P-ATP is the method of our choice for sequencing analysis. One cDNA template was prepared by RT-PCR technique starting from the 5'-end nucleotide 1-465 of the dengue-3 genome. In our cycle sequencing experiments, the substitution of 7-deaza-dG was used for dG in DNA eliminated much of the secondary structures that produced gel artifacts. The final sequence result of this cDNA template was established from its sequence data determined on both strands in opposite directions. Alignment between the newly established nucleotide sequence as well as its deduced amino acid sequence of the Bangkok dengue-3 virus and the published sequence data of the dengue-3 prototype (H87) was manipulated by the PC-DOS-GIBIO-DNASIS TM 06-00 (Hitachi Software). According to the deduced amino acid sequence of the Bangkok dengue-3 virus, its C protein was found to be highly positively charged because of large numbers of lysine and arginine. The homology of the nucleotide sequence between the two dengue-3 virus revealed 97%. The deduced amino acid sequences from the nucleotides 95-465 of the two viruses showed the same indicating highly conserved capsid proteins. Multiple alignment of the nucleotide sequences as well as the deduced amino acid sequences among the Bangkok dengue-3 virus and other dengue 3 viruses also confirmed the highly conserved 5'-noncoding regions and the capsid proteins. PMID- 11414441 TI - A novel method for the preparation of large cDNA fragments from dengue-3 RNA genome by long RT-PCR amplification. AB - For many years, dengue viruses were among the most difficult flaviviruses to isolate and to identify, but technical advances in the past 20 years have facilitated this process. Dengue viruses are usually recovered from specimens by the infection of mosquito-cell cultures. The virus may be passaged several times in cell cultures until a sufficient infectivity titer is attained. The viral nucleocapsid consists of capsid protein and an RNA genome. The dengue genome is a single stranded messenger (positive) sense RNA of approximately 11 kb in length. The isolation of dengue genomic RNA from various sources requires precautions to avoid RNases. RNases are released during cell disruption, and their activity must be inhibited as quickly as possible by using guanidinium thiocyanate in the presence of 2-mercaptoethanol. There has recently been a revolution in molecular biology with the development of the powerful reverse transcriptase (RT) and polymerase chain reaction (PCR) technology. Advanced studies on RT technique lead to much further improvement of the reverse transcriptase enzyme by genetic engineering. The Superscript II RNase H- RT (GIBCO BRL, USA) is genetically engineered DNA polymerase that synthesizes a complementary DNA strand from single stranded RNA. DNA or an RNA-DNA hybrid. This enzyme is produced from a cloned M MLV RT gene constructed by the introduction of point mutation in the RNase H active center. The selective mutations within the RNase H domain maintain full polymerase activity. This structural modification eliminates degradation of RNA molecules during the first strand cDNA synthesis. The combination of thermostable DNA polymerase with and without proofreading activity (3'-exonuclease activity), improved buffer conditions and thermal cycling profiles overcome the length limitation of PCR. On the basis of these findings, we have developed a long RT PCR system for preparing large cDNA fragments of dengue 3 virus (H-87) by using the Superscript II RNase H- RT for reverse transcription and a mixture of Taq and Pwo DNA polymerases for PCR. Three large cDNA fragments covered the full genomic RNA from the 5'-end to the 3'-end of dengue-3 virus (H-87; 10,696 bps) could be successfully prepared as the lengths of 2.437 bps, 3,980 bps and 4,337 bps respectively. The ability of our developed long RT-PCR will bring speed and simplicity to genomic mapping and sequencing and facilitate studies in molecular genetics of dengue viruses. PMID- 11414442 TI - The risk of dengue hemorrhagic fever (DHF) outbreak based on vector density in Kurau, Riau province, Indonesia. AB - It is known that in Padang, Rantau, Rangsang, Merbau and Bengkalis islands, Riau Province, the deposit of oil was found in a huge quantity. The drilling concession belongs to Kondur Petroleum Company. To operate an exploitation, hundreds of workers not only Indonesian but also the workers from foreign countries come and go to that area. It was recorded that the workers from foreign countries come from Singapore, Malaysia, Thailand, The Philippines, Taiwan, Japan, Korea. United States of America and from France, Britain, Australia and Germany. These workers have a close interaction with about 50,00 local population distributed with a high concentration in some places. The high risk of DHF was determined since the significant density of Aedes aregypti larvae, the main vector of DHF, were found in Lukit, Mengkikip and Melibur, three locations of survey. Of 104 (53.3%) of 195 houses in these three survey locations were found positive for the larvae. Even though there were no positive of larvae in 65 houses in Lukit, in Melibur and Mengkikip, the House Index (HI) was 61.7% and 95.7%. Outside the houses 521 containers were examined in three locations of survey and 329 (63.1%) were positive for Ae. aegypti larvae. The highest number of containers positive for Ae. aegypti larvae were 213 (94.7%) out of 225 and found in Mengkikip. In Melibur, 114 (68.3%) out of 167 of containers were positive and in Lukit only 2 (1.56%) out of 129 containers were positive of Ae. aegypti larvae. These larvae density constitute a high risk of DHF outbreak, and unfortunately is supported by the rainfall situation recorded in Kurau and BZ Climatology Stations. It was recorded that all along the year, at least one day in a month there was the rain which the rainfall volume was 30 ml. PMID- 11414443 TI - Clinical presentations and outcomes of TB among HIV-positive patients. AB - A retrospective cohort study reviewed the hospital records of 200 consecutive HIV patients and followed for 6 months period of TB treatment. Sociodemographic details, symptoms, and signs, results of investigations were recorded at the time of presentation, while diagnosis, and clinical outcomes were done at the end of the follow up period. The aim of the study was to determine the clinical manifestations and outcomes in tuberculosis among HIV-positive patients in Bamrasnaradura Hospital, Nonthaburi. Also, predictive factors for loss of follow up in these patients were determined. The results showed that extrapulmonary TB (58%) was more common, and lymphadenopathy (52%) was the most common sign on examination. Chest x-rays were positive in only 55% cases, while AFB examinations were positive in 48.5%. After 6 months, 30% patients were alive, 12% patients died, and 50% patients were lost to follow up. Factors such as low socioeconomic status (p<0.001), past history of TB (p<0.003) etc were statistically significant in predicting the likely loss of follow up and thus, nonadherence in TB management among HIV- positive patients. In conclusion, tuberculosis in HIV patients presents as extrapulmonary involvement more frequently. The diagnosis is more difficult in cases of HIV. Patients' noncompliance is the biggest problem in management of TB in these patients, but there are predictors which can help us identify the likely patients, and thus, improve their management and clinical outcomes. PMID- 11414444 TI - Human T-lymphotropic retrovirus type-1 in Thailand. AB - Seroprevalence of antibodies to human T-lymphotropic virus type-1 (HTLV-1) was surveyed among the Thai population by the particle agglutination test and Western blotting test. None of a total of 727 individuals from seven ethnic groups were positive for the specific antibody to HTLV-1. Among hospital based 3,427 subjects in Southern Thailand, one patient with a brain tumor showed positivity in the Western blotting test, however, HTLV-1 proviral genome was not identified by PCR. The present data suggest that HTLV-1 is not endemic in the Thai population and that HTLV-1 is not a major public health problem in Thailand because HTLV-1 rarely causes its associated diseases. PMID- 11414445 TI - Antigenic heterogeneity of lipopolysaccharide among Burkholderia pseudomallei clinical isolates. AB - Burkholderia pseudomallei (BP) causes melioidosis, a potentially fatal human infection in the tropics. Clinical isolates from different geographical locations have similar morphological and biochemical characteristics. Although BP has been reported to possess 2 types of lipopolysaccharide (LPS) differing in the chemical structure of their O-polysaccharide (O-PS) component, earlier report demonstrated that the clinical strains exhibited identical LPS moieties. Recently, we reported antigenic similarity between the pathogenic (Ara-) and nonpathogenic (Ara+) biotypes. However, a few clinical isolates showed atypical SDS-PAGE profiles. In this study, LPS from 739 BP isolated from patients and animals in different geographical areas were extracted by proteinase K digestion method. Their SDS PAGE profiles and their immunoreactivities with patients' sera and monoclonal antibody (MAb) to LPS were analyzed. The isolates showed 3 LPS patterns differing in the number and electrical mobility of bands in silver-stained gel. A majority of BP (711) isolates exhibited identical typical ladder pattern, 21 isolates showed atypical ladder pattern and 7 isolates did not exhibit ladder appearance. However, all LPS preparations exhibited similar endotoxic activity as determined by Limulus amebocyte lysate assay. On the other hand, there were no immunological cross reactivity between typical and atypical LPS, as judged from Western blot against homologous and heterologous sera from melioidosis patients from whom the typical and atypical LPS were isolated. Nevertheless, a Western blot profile of the typical LPS showed some variations when probed with MAb against BP LPS (9D5). Heat-killed bacteria from all LPS groups could similarly activate mouse macrophage cell line to produce nitric oxide (NO) and inducible NO synthase (iNOS). PMID- 11414446 TI - Prevalence of enterotoxigenic motile aeromonads in children, fish, milk and ice cream and their public health significance. AB - Motile aeromonads, which have been implicated as causative agents of human gastroenteritis, meningitis. endocarditis and osteomyelitis, were isolated from fish, pasteurized and flavored milk and ice creams. Forty-five isolates of Aeromonas from diarrheic children (100) and 285 food samples comprising of fish (100), milk (85) and ice creams (100) were identified as A. hydrophila (21), A. sobria (16) and A. caviae (8). Five of these 45 isolates of Aeromonoas (3 A. sobria, 2 A. hydrophila) were from fecal samples of diarrheic children under five years of age. Twenty-six of these 45 isolates were found to be enterotoxin producing by ligated rabbit ileum loop technique. Isolation of enterotoxigenic motile aeromonads from food as well as diarrheic children have great public health significance which will be further discussed. PMID- 11414447 TI - Mechanical carrier of bacterial enteric pathogens by Chrysomya megacephala (Diptera: Calliphoridae) in Chiang Mai, Thailand. AB - Chrysomya megacephala was studied regarding its mechanically bacterial carrier in urban areas of Chiang Mai, northern Thailand. Fifty-six adult flies were randomly collected using sweep insect net during April-May, 1999 from 3 fresh food markets and examined for bacteriological isolation. Among them, 49 flies (87.5%) were bacterial carriers. The total 22 bacterial species and 8 groups were isolated. Three species previously reported as the bacterial enteric pathogens causing diarrheal disease were isolated from 5 flies, ie Aeromonas hydrophila, Edwardsiella tarda and Vibrio cholerae non-01, with their prevalence rates in flies being 3.579, 1.79% and 3.57%, respectively. Five possible bacteria enteric pathogens, ie Aermononas sobria, Citrobacter freundii, Escherichia coli, Providencia alcalifaciens and Pseudomonas aeruginosa, were isolated from 21 flies with the prevalence rates in flies being 5.34%, 3.57%, 26.79%, 7.14% and 1.79%, respectively. The bacterial load isolated from all 3 pathogenic species was entirely found more than 10 colony per fly, indicating the high chance for disease transmission via this fly species. C. megacephala may play the possible and/or important role of bacterial enteric pathogens transmission, thereby promoting the public health personnel for sanitation improvement in fresh food markets and fly control management in these particular areas. PMID- 11414448 TI - Neurocysticercosis and echinococcosis in Asia: recent advances in the establishment of highly reliable differential serodiagnosis for international collaboration. AB - Neurocysticercosis (NCC) and echinococcosis, caused by the larval stage of taeniid cestodes, are recognized as major parasitic zoonoses threatening human life worldwide. Cystic echinococcosis (CE), caused by Echinococcus granulosus, has well been known to be more widely distributing in Europe and Asia (Eurasia) than alveolar echinococcosis (AE) caused by E. multilocularis. However, it has recently been found that AE is more widely distributing or spreading in Eurasia. Furthermore, NCC caused by Taenia solium is also spreading in Eurasia. Due to the lack of reliable methodology for diagnosing these zoonotic cestodiases worldwide, prevalence rates of these diseases are extremely underestimated. Our group has been working for the establishment of differential serodiagnosis and molecular diagnosis of AE. CE and NCC as international collaboration projects sponsored by the Ministry of Education, Japan from 1994 until 2000 at least. In this paper, we introduce (1) the most recent original work on the establishment of differential serodiagnoses of NCC, AE and CE, (2) international collaboration work on epidemiology of these diseases in several countries, and discuss (3) what we can and should do for the control of such global parasitic diseases. It is stressed that international collaboration or cooperation work on the control of parasitic diseases is only successful based on the original scientific contribution of high standard. PMID- 11414449 TI - in children. Its results were drawn with limited sample size obtained after a short phase of data collection. A further study should be conducted to get more precise and valid results that may be useful for preventing UTI among children. AB - This hospital-based case-control study was conducted from September 1998 to January 1999 in Metro Manila, Philippines. General objective of the study is to determine the association between selected hygiene behavior and urinary tract infection (UTI) among children aged 6-12 years. Specifically, the study is designed to examine the relationship between UTI and urination, defecation, washing and bathing habits. Twenty-three cases of children with UTI and an equal number of controls were recruited in four tertiary hospitals. The study association was determined by using odds ratio, the chi-square test and the Fisher exact test, where appropriate, in simple analysis. Furthermore, exact logistic regression analysis was applied to overcome the problem of small sample size. The data suggested that bathing habit less than daily, holding of urination during daytime, and washing habit after defecation might have risk effects on UTI. There was not enough evidence of significant association between UTI and other study exposures. Among extraneous variables, age group or school enrollment of children had a borderline significant association with UTI after adjusted simultaneously for selected variables. This study served as a pilot of the Preventive Nephrology Project (Department of Health, Philippines) in determining selected risk factors of PMID- 11414450 TI - TNF-alpha induces caspase 3 (CPP 32) dependent apoptosis in human cholangiocarcinoma cell line. AB - Cholangiocarcinoma (CCA), a malignant tumor derived from bile duct epithelium, occurs with a higher incidence in tropical countires especially in some areas of Southeast Asian countries such as Thailand. This tumor is relatively resistant to chemotherapy. In this study, molecular mechanism of killing of this tumor by TNF alpha was investigated. Human cholangiocarcinoma cell line (HuCCA-1) was developed and used as a model for treatment. Activation of HuCCA-1 with TNF-alpha in the present of actinomycin D (1 microg/ml) caused death of the tumor cells. Western blotting analysis of the cells extracted demonstrated the cleavage of poly (ADP-ribose) polymerase (PARP) within 6-8 hours following TNF-alpha treatment indicating apoptotic death. The cleavage of PARP was inhibited when the cell line was pretreated with peptide inhibitor, Ac-DEVD-CHO, suggesting that apoptosis induced by TNF-alpha of this cell line involves activation of caspase II subfamily. The procaspase 3 (proCPP-32), one of the caspase group II subfamily was degraded after the HuCCA- I cell line was treated with TNF-alpha. Furthermore, Gelsolin, an 83 kDa protein which is identified as caspase 3 substrate, was cleaved to 43 kDa fragments after the cells were treated with TNF alpha. These results indicate that apoptosis of human cholangiocarcinoma cell line as induced by TNF-alpha treatment is mediated through caspase 3. PMID- 11414451 TI - Dominant lethal test in rats treated with some plant extracts. AB - The present study was undertaken to investigate the toxic effect of aqueous extracts of Aegle marmelos (AM), Stevia rebaudiana (SR), Pouteria cambodiana (PC) and Clausena excavata (CE) on rats by dominant lethal test. The data of 8-week treatment suggested that none of the extracts adversely affected male body and testicular weights as well as cauda epididymal sperm counts. No notable changes in sperm morphology and motility were observed. On the other hand, sperm count in the CE group was significantly higher as compared to both control and other treatment groups. There were no abnormal changes in the number of implantation sites, number of viable fetuses and number of dead fetuses in females mated with plant extract-treated males relative to controls. Based on these results, it could be concluded that all the investigated plant extracts have no toxic effect on male rat reproduction and progeny outcome. PMID- 11414452 TI - Anti-headlice activity of a preparation of Annona squamosa seed extract. AB - Various parts of Annona squamosa Linn. (custard apple) have long been used in Thai folk medicine. The effectiveness of organic solvent extracts of custard apple seeds and leaves against headlice has been reported. The present study is focused on the efficacy and stability of seed extract prepared as a cream. A petroleum ether extract of seeds was prepared as a 20% w/w oil in water cream and tested for anti-headlice activity in school girls. After a 3 hour-hair exposure to 20 g of freshly prepared custard apple cream, 95.34+/-1.96% of headlice were killed. This killing rate was higher than that obtaned from the control (cream base) or the standard drug (25% benzyl benzoate emulsion). The anti-headlice efficacy of the 6 and 12 month-storage preparations kept in a refrigerator and 12 month-storage at room temperature were 99.19+/-0.60%, 97.55+/-2.06% and 89.47+/ 3.64%, respectively, which were not significantly different from that of the freshly prepared cream. This cream did not cause any irritant effect on the scalp or neck skin while benzyl benzoate emulsion produced burning sensation or redness. The results demonstrate that the petroleum ether extract of Annona squamosa seeds prepared as a cream is stable for at least 12 months. The custard apple cream may be, therefore, suitable for use as an alternative therapy against headlice. PMID- 11414453 TI - Antifungal activity of Curcuma longa grown in Thailand. AB - Curcuma longa Linn. or turmeric (Zingiberaceae) is a medicinal plant widely used and cultivated in tropical regions. According to Thai traditional texts, fresh and dried rhizomes are used as peptic ulcer treatment, carminatives, wound treatment and anti-inflammatory agent. Using hydro distillation, 1.88% and 7.02% (v/w) volatile oils were extracted from fresh and dried rhizomes, respectively, and 6.95% (w/w)crude curcuminoids were extracted from dried rhizomes. Dried powder was extracted with 95% ethanol and yielded 29.52% (w/w) crude ethanol extract composed of curcumin (11.6%), demethoxycurcumin (10.32%) and bisdemethoxycurcumin (10.77%). These extracts were tested for antifungal activity by agar disc diffusion method against 29 clinical strains of dermatophytes. It was found that crude ethanol extract exhibited an inhibition zone range of 6.1 to 26.0 mm. There was no inhibition activity from crude curcuminoids while curcumin, demethoxycurcumin and bisdemethoxycutcumin gave different inhibition zone diameters ranging from 6.1 to 16.0 mm. Although antifungal activity of undiluted freshly distilled oil and 18-month-old oil revealed some differences, the inhibition zone diameters for both extracts varied within 26.1 to 46.0 mm. With 200 mg/ml ketoconazole, the activities of the standard agent were similar to the oil, both freshly distilled and 18-month-old, but were significantly different from those of curcuminoid compounds and crude ethanol extracts (p < 0.01). Turmeric oil was also tested for its minimum inhibitory concentration (MIC) by broth dilution method. The MICs of freshly distilled and 18-month-old oils were 7.8 and 7.2 mg/ml respectively. PMID- 11414454 TI - Neurocysticercosis: utilizing the cystic fluid antigen from Taenia solium metacestodes for diagnosis by IgG-ELISA. AB - Cystic fluid, which has antigenic properties of whole Taenia solium cysticerci, was used to discriminate neurocysticercosis cases and other parasitic infections, especially helminthiases. Twenty-one neurocysticercosis and several kinds of 22 different parasitic infections, including HIV cases (n=234) evaluated a 90.48% sensitivity and 86.32% specificity of indirect ELISA as follows: a low antigen concentration of 5 microg/ml. serum dilution of 1:400, conjugate dilution of 1:2,000 and a cut-off value of 0.349. Eight different helminthic infections (n = 25); echinococcosis (8/10), gnathostomiasis (6/8), strongyloidiasis (5/14), hookworm infection (1/18), angiostrongyliasis (2/25), opisthorchiasis (1/18), onchocercosis (1/3) and toxocariasis (1/6) were cross-reactive with this antigen. No serum antibody from other brain infections in the study gave a reaction with the antigen. In this study, the cystic fluid antigen gave high sensitivity of the test. However, the antigen contains various antigenic molecules able to bind with antibodies from several of the above helminthic sera, especially echinococcosis and gnathostomiasis. In Thailand, gnathostomiasis is one of the more famous tropical diseases but echinococcosis is quite rare. Cystic fluid antigen should be further investigated for its specific finding in diagnosis. PMID- 11414455 TI - Epidemiology, symptoms and treatment of paragonimiasis in Sin Ho district, Lai Chau province, Vietnam. AB - Sin Ho is a district of the northern mountainous province of Lai Chau, Vietnam, where the people have the habit of eating undercooked crabs. A study on paragoniamiasis carried out from 1994 to 1995 with 1,642 persons in this endemic area, showed that the rate of eating raw-crab was 72.5%. Crab examination (Ranguna kimboiensis) showed an infection rate of Paragonimus metacercaria of 98.1%. With 624 stool samples examined by Kato technique, the infection rate of Paragonimus in humans was 6.4%. With 338 sputum samples examined by direct and centrifuge methods, the infection rate of Paragonimus was 7.4%. Most of the patients were children (63.2%). The infection rate of Paragonimus in dogs was 18.2 - 33.3%. Adult worms, collected from the dogs in the field and from the cats in laboratory, were identified as Paragonimus heterotremus. The main symptoms of Paragonimus patients were cough and hemoptysis (92%), discontinuously developed (96%), without fever (94%), chest pain (70%), pleural effusion (26%), neurogical symptoms (8%), eosinophilia (88.9%), nodular ring shadows in the lungs, as shown by chest X-ray examination and more in lower lobe, (76.2%). Paragonimiasis patients were treated by (a) Praziquantel 25 mg/kg/day x 3 days; the cure rate was 68.8%. (b) Praziquantel 50 mg/kg/day x 3 days; the cure rate was 75%. PMID- 11414456 TI - The karyotype of the lung fluke, Paragonimus siamensis. AB - The karyotype of chromosomes obtained from the germ cells of Paragonimus siamensis was analyzed by air-drying technique. The result revealed the diploid number of chromosomes to be 2n=22. The entire chromosomes consisted of one large sized metacentric, four medium-sized subtelocentrics, three small-sized metacentrics or submetacentrics and three small-sized submeta-centrics or subtelecentrics. The relative arm lengths of the chromosomes were 20.13+/-1.21, 13.16+/-0.42, 11.07+/-0.48, 10.37+/-0.84, 9.29+/-0.52, 6.50+/-0.39, 6.42+/-0.34, 6.27+/-0.28, 6.04+/-0.58, 5.65+/-0.39 and 5.03+/-0.40% respectively. These data were comparable with those of other related species. PMID- 11414457 TI - Comparative studies on protein, isoenzyme and DNA restriction endonuclease profiles of Thai Paragonimus metacercariae. AB - General proteins and 14 enzymes from metacercariae of Paragonimus heterotremus, P. siamensis and P. westermani were determined by vertical polyacrylamide gel electrophoresis. The isoenzyme profiles showed considerable interspecific polymorphism for general protein (PT), malate dehydrogenase (MDH), malic enzyme (ME) and tetrazolium oxidase (TO) while those of glucose phosphate isomerase (GPI) and phosphoglucomutase (PGM) showed similarity. The Pt-6 and To-I loci can be used as identification markers for these three species. The preliminary study of the molecular biology of Paragonimus heterotremus P. siamensis and P. westermani was based on analysis of metacercarial genomic DNA with restriction endonuclease Pst I. Agarose gel electrophoresis revealed restriction fragment length differences among the three species studied. The DNA restriction fragments were approximately 4-6 fragments, ranging from 5.35 to 14.67 kb. Among these. P westermani shared two homologous fragments with P. siamensis, ie, 5.35 and 7.22 kh, none with P. heterotremus, while P. heterotremus shared only one with P. siamensis, ie, 8.16 kb. Thus, the DNA restriction fragment length differences can be used to differentiate among these three species. PMID- 11414458 TI - Sarcocystis spp infection in Philippine water buffaloes (Bubalus bubalis). AB - In a survey of sarcocysts in muscle tissues obtained from 142 water buffaloes, 65% of the carcasses had sarcocysts. Macroscopic and two forms of microscopic sarcocysts, the spindle-shaped or fusiform sarcocysts commonly occurring in the muscles of the esophagus, throat and limbs, and the globular to oval-shaped sarcocysts which were the dominant form in the diaphragm and cervical muscle tissues were noted. Ultrastructural analysis of macroscopic and microscopic sarcocysts and their cyst wall revealed two distinct species of Sarcocystis: the macroscopic species, Sarcocystis fusiformis which has been previously reported in Philippine carabaos possessing highly dendritic cauliflower-like projections emanating from the primary cyst wall, with annulated microfilaments and numerous electron dense granules: and the Sarcocystis levinei (Dissanaike and Kan, 1978) Huong, Dubey and Uggla. 1997 exhibiting a cyst wall with undulating and hair-like villar protrusions with expanded or dome-shaped base, intermediate finger-like, and distal tapering segments which at some points join to form conical tufts. Our findings represent the first report of S. levinei in the country supported with ultrastructural analysis of the sarcocysts and cyst wall, and likewise refute earlier published reports that all microscopic sarcocysts in Philippine carabaos are developing forms of the macroscopic species, S. fusiformis. Histopathological changes such as displacement and necrosis of the surrounding host muscle tissue were observed with macroscopic sarcocysts and histologically processed tissue samples containing microscopic fusiform sarcocysts. Necrotic myofibrils and mitochondria were evident in ultrathin sections. PMID- 11414459 TI - Angiostrongyliasis: analysis of antigens of Angiostrongylus costaricensis adult worms versus IgG from infected patients with Angiostrongylus cantonensis. AB - The possibility of cross-reactivity was previously investigated by indirect ELISA with sera from Angiostrongylus cantonensis infections, normal controls and A. costaricensis antigen. 5 microg/ml of crude antigen from both sexes of each species reacted with diluted serum samples (1:800) of each of 20 cases of angiostrongyliasis and normal controls, and further with anti-human IgG conjugate at 1:1,000. The mean absorbance values were evaluated as follows; normal controls showed a value of 0.033 using A. costaricensis antigen lower than (0.085) A. costaricensis antigen. Both mean values of angiostrongyliasis cases were rather close (0.491) using A. costaricensis antigen and the other antigen (0.518). The present study continued with a crude antigen of 13 A. costaricensis females and males. Serum samples were analyzed; 27 sera of angiostrongyliasis, 30 negative controls and 193 cases of other parasitic infections (91 cases of nematodiasis; 45 cases of cestodiasis; 47 cases of trematodiasis and 10 cases of HIV) and 7 cases of other brain infections. This antigen was evaluated for ELISA with a concentration of 5 microg/ml, serum dilution 1:400 and anti-human IgG conjugate at 1:2,000. The test gave sensitivity and specificity at cut-off value 0.261; 92.59% and 73% respectively. The antigen was cross-reactive with 30 cases from 9 out of 10 different kinds of nematodiasis (gnathostomiasis, strongyloidiasis, ascariasis, hookworm infections, trichinosis, toxocariasis, trichuriasis, onchocercosis and Wuchereria bancrofti infections. Five cases from 3 of 6 kinds of cestodiasis (neurocysticercosis, echinococcosis and Hymenolepis nana infections) and 18 cases of 4 out of 5 kinds of trematodiasis (Paragonimus heterotremus infections, opisthorchiasis, schistosomiasis and fascioliasis). One case of other brain infections was observed. The crude antigen of A. costaricensis showed a high percentage sensitivity with serum antibodies of angiostrongyliasis cases. Low specificity of the test was observed by reactions of those serum antibodies with various kinds of antigenic molecules. This study provides baseline data for further immunodiagnosis of human angiostrongyliasis. PMID- 11414460 TI - Metacercaria from freshwater fishes of Mae Sa stream, Chiang Mai, Thailand. AB - Metacercaria from freshwater fishes of Mae Sa stream, Doi Suthep-Pui National Park. Chiang Mai Province were investigated between January 1997-December 1998, 3,133 fishes of 32 species were collected. The prevalence of infection of fishes was 19.087% (598/3, 133). Six genera of metacercaria were recovered as follows: Acanthostomum sp, Centrocestus caninus, Haplorchis taichui, Haplorchoides sp, Posthodiplostomum sp and Stellantchasmus falcatus. The prevalence of each species of metacercaria was observed from total number of fishes as follow: 1.149% (36/3,133), 3.753% (96/3,133), 1.883% (59/3,133), 9.735% (305/3,133), 0.096% (3/ 3,133) and 2.553% (80/3,133) respectively. The highest prevalence of parasites were observed: Acanthostomum sp was 11.111% from Trichopsis vittatus; Centrocestus caninus was 100% from Xenentodon cancila; Haplorchis taichui was 12.108% from Mystacoleucus marginatus; Haplorchoides sp was 55.532% from Mystacoleucus marginatus; Posthodiplostomum sp was 1.176% from Dermogenus pusillus; Stellanchasmus falcatus was 100% from Xenentodon cancila. Centrocestus caninus, Haplorchis taichui and Stellantchasmus falcatus have been reported from human and mammals. The rest were reported from birds, fishes and reptiles. PMID- 11414461 TI - IgG- and IgG4-detected antigens of Dirofilaria immitis adult worms for bancroftian filariasis by enzyme-linked immunoelectrotransfer blot. AB - In Thailand, Wuchereria bancrofti filariasis has persisted along the border between Thailand and Myanmar, its dynamic distribution caused by the infected transmigrants between neighboring countries, and the availability of susceptible mosquito vectors. Dirofilaria immitis adult worm was used as a source of antigens, excretory-secretory (ES) and partial surface extracts, to detect human filariasis. ES products showed several stained bands with Coomassie brilliant blue ranging from 14.5-93 kDa and mostly being glycoproteins as shown by concurrent reaction with Concanavalin A, except those at 18, 16 and 14.5 kDa which stained only with Coomassie brilliant blue. Surface proteins of 33.5-91.5 kDa were stained with Coomassie brilliant blue and showed smear bands with Concanavalin A. By enzyme-linked immunoelectrotransfer blot, Bancroftian filariasis sera gave specific reactions with glycoprotein ES antigens at MW 20.5 kDa against anti-human IgG. A prominent band of 18 kDa appeared consistently with the IgG4-ES antigen system. Surface extracts reacting with IgG and IgG4 were considered to be unsuitable as antibodies from all cases of filariasis could not detect any bands. PMID- 11414462 TI - Reinvestigated life history of Moniliformis dubius Meyer, 1933 in Chiang Mai, Thailand. AB - The lite cycle of Moniliformis dubius Meyer, 1933 was investigated by using rats (Rattus norvegicus) as the definitive host and American cockroach (Periplaneta americana) as an intermediate host. At room temperature (22-26 degrees C), the shelled acanthor hatched in mid-gut of P. americana 3 days postinfection. released to be the acanthor and then penetrated the gut wall into the hemocoel 5 days postinfection. It further developed into acanthella with proboscis primordium 16 days postinfection. The cystacanth stage recovered 45-52 days postinfection. Rats were fed with the cystacanth, their intestine and feces were examined daily. Shelled acanthor was found in feces on the 31st day of the infection. From the above results, the life cycle of M. dubius was completed within 76-83 days. Observation of the mature adult by scanning electron microscope, both sexes were similar in morphology. PMID- 11414463 TI - Review and recent progress: the mathematical modeling of mixed species Plasmodium infections. AB - Mathematical modeling serves numerous uses in biology, notably in the exploration of phenomena that are difficult to observe empirically. Here, I review recent progress in modeling the blood-stage dynamics of mixed-species Plasmodium infections, namely P. malariae-P. falciparum and P. vivax- P. falciparum mixed infections. Modeling reproduces features of such infections found in nature including the asymmetry of parasite blood-stage densities, inter-specific suppression, and parasite asexual-form recrudescence following long-standing sub patency. Several findings which merit clinical attention are presented: the ability of P. malariae and P. vivax to reduce the peak parasitemia of co infecting P. falciparum, and the potential recrudescence of a low-level P. falciparum infection following a P. malariae infection or P. vivax infection or relapse. The action of antimalarial drugs is discussed, highlighting some potential complications in treating mixed-species malaria infections. Most notably, if a mixed-species infection is misdiagnosed as a single-species P. vivax infection, treatment can lead to the dangerous appearance of "hidden" P. falciparum. PMID- 11414464 TI - Factors influencing blood viscosity: adult and newborn blood analysis. AB - Our finding of a decrease in blood viscosity in newborn infants compared with adults leads to analysis of this change by measuring blood viscosity, plasma fibrinogen concentration, plasma viscosity and mean corpuscular volume (MCV) of 24 adults and 16 newborn infants. Plasma proteins in newborn infants are synthesized to a lesser extent than in adults, therefore plasma fibrinogen concentration of newborn infants is less than that of adults. This causes decreased blood viscosity in newborn infants compared with adults. Secondly plasma viscosity of newborn infants is also less than that of adults. In the same situation this causes a decrease in blood viscosity in newborn. Thirdly. MCV of newborn infants is greater than that of adults because of young red cells. There is a negative correlation between blood viscosity and MCV. High MCV in newborns leads to low blood viscosity in newborn infants, compared with adults. PMID- 11414465 TI - Application of yeast enolase as antigen for immunodiagnosis of malaria. AB - In 1998, we reported that Plasmodium falciparum (Pf) enolase was useful as the capture antigen for the immunodiagnosis of malaria. In the present study, we modified a fluorescence-ELISA for the diagnosis of malaria by applying yeast enolase or rabbit muscle enolase as antigen. Sera from 67 falciparum malaria patients and 15 vivax malaria patients were tested by the method. Positivity rates of the former was 82.1% against yeast enolase antigen and 90.5% against rabbit muscle enolase antigen, and those of latter was 93.3% against both enolase antigens. Mean antibody level (RFU values) of sera from falciparum and vivax malaria patients were significantly higher than those from healthy individuals. There was a significant correlation between anti-yeast and anti-rabbit muscle enolase antibody level (RFU values) in the group of falciparum subjects (r = 0.401, p<0.001). A significant correlation between RFU values against yeast enolase antigen and indirect fluorescent antibody titers against crude Pf antigen in the same subjects was recognized (r = 0.518, p<0.001). Longitudinal changes of RFU values against yeast enolase for the following 4 weeks after admission were also examined for sera from falciparum malaria patients. Patients with more severe malaria showed increasing RFU values as the clinical courses progressed. However, in the mild cases, each RFU value stayed unchanged during the course. We concluded that yeast and rabbit muscle enolase could be appropriately used as antigen for the immunodiagnosis of malaria. PMID- 11414466 TI - Autoclaved cercarial vaccine against schistosomiasis: ultrastructural and biochemical studies. AB - Autoclaved cercarial vaccine (ACV) was found to be highly effective in eliciting protective immunity against experimental Schistosomal mansoni. So, the aim of this study was to analyse ACV biochemically and to study ultrastructural changes inflicted on the cercariae as a result of autoclaving, thus rendering it highly protective. Results of this study showed that approximately 100 microg protein and 44 microg carbohydrate were obtained from 10(3) cercariae. The predominant sugar was fucose. Galactose, glucose, manose, galactosamine and glucosamine were also detected. Threonine, glycine, serine and glutamic acid comprised approximately 53.7% of the amino acid residues of the protein. Ultrastructural study revealed preserved architecture of the cercariae. The tails were still attached to the posterior ends of the bodies. However, in others the tails were separated from the bodies and appear schistosomula like. There were also some morphological changes such as thinning of the pericortical envelop with appearance of surface pores. PMID- 11414467 TI - Clinical presentation of childhood malaria in Savannakhet province, Lao PDR. AB - A descriptive study on the clinical presentation of childhood malaria was conducted in Savannakhet Province, Lao People's Democratic Republic. It is aimed to describe the clinical features and to determine the association between the severity of malaria and the initiation or delay of treatment. A total number of 92 children 1-14 years of age with confirmed malaria diseases were enrolled in this study. Fifty-six cases (60.9%) had illness for less than 3 days before hospitalized and 36 cases (39.1%) for more than 3 days. Twenty-nine cases (31.5%) had self antimalarial medication before admission (9 cases of chloroquine, 16 cases of quinine and 4 cases of artesunate). Ten cases (10.9%) had abnormal consciousness of which 7 cases (7.6%) had confusion but responded to verbal command and 3 cases (3.3%) were in coma not respond to painful stimuli but had reflex. Two cases 2.2%) had convsulsions, 11 cases (12.0%) had dehydration, 47 cases (51.1%) had vomiting, 18 cases (19.6%) had hepatomegaly and 19 cases (20.7%) had splenomegaly. There was a statistically significant association between consciousness levels and the duration of illness before admission < or = 3 days and > 3 days (p = 0.01) while there is no significant difference between parasitemia density and the duration of illness before admission (p > 0.05). PMID- 11414468 TI - A study of anemia in pregnant women with Plasmodium falciparum at district hospitals in Vientiane, Lao PDR. AB - A descriptive cross-sectional study was carried out among pregnant women attending antenatal care at the district hospital, with suspected clinical manifestation of malaria in order to determine the prevalence of anemia and malaria among pregnant women and to determine any correlation between degree of anemia and degree of malaria parasitemia in pregnancy with malaria infection. This is a quantitative research method using face-to-face questionnaire. This study was undertaken at the district hospitals of Vientiane Prefecture and Vientiane Province. Sixty-eight pregnant women with suspected malarial clinical manifestations attending the antenatal care at these hospitals were recruited during June - October, 1998. The subjects were asked about their sociodemographic, socio-economic characteristics, gravida and parity, gestational age, last pregnancy and past history of hematology diseases. Blood samples (dry smear for thick and thin blood films) were examined at the same time for Plasmodium falciparum. The study showed that the prevalence of anemia (Hb < 11 g/dl) and severe anemia (Hb 4-6.9 g/dl) in the total sample population was 48.5% and 8.8% respectively. However, the prevalence of anemia among pregnant women with malaria was 68.75% compared to those without malaria infection (42.31%), but the difference was not statistically significant (p=0.117). A plausible explanation could be small sample size. The prevalence of severe anemia in pregnancy with malaria parasitemia was 18.8% compared to those without parasitemia (5.8%). The difference was not statistically significant (p=0.102). The difference of the mean hemoglobin level in falciparum positive cases and falciparum negative cases was clinically and statistically significant (RR = 1.63 and p=0.00679). There was some evidence of a negative correlation between the degree of anemia and parasitemia count (r= -0.19 and r2= -0.04). In conclusion this population had high prevalence of anemia in pregnant women and P. falciparum may be the main factor associated with anemia. There is a need to investigate other causes of anemia among pregnant women. Our results suggest that frequent and regular antenatal monitoring is necessary for the pregnant women. They should be encouraged to attend antenatal clinics through health education, increased health personnel awareness of proper management for the pregnant women with fevers from malarial endemic areas. There is a need for further research in this area in order to obtain adequate sample size. PMID- 11414469 TI - The situation of malaria along the Vietnam-Lao PDR border and some related factors. AB - This was a descriptive cross sectional study. It was done in 4 communes along the Vietnam-Lao PDR border of two mountainous provinces: Sonla and Nghean. The cluster multistage sampling technique was applied to choose the study sites. The results of the study show: Among the 2,441 persons given blood tests to find malaria parasites, 0.7% of them carry malaria parasite, of whom 0.6% carry P. falciparum and 0.1% carry P. vivax. The malaria morbidity in the year was 6.9%. The mortality due to malaria is 1.59 per 100,000 population per year. Among the 106 hamlet motivators being interviewed, only 75.5% knew that malaria is transmitted by mosquitos, 71.7% knew that malaria patients are a source of transmission, over 50% of the motivators have mistaken understanding about the living environment of malaria mosquitos. Most of them have had mistakes in diagnosis, treatment of malaria, mosquito-killing spraying. Among the 729 adults being interviewed, 59.0% did not know about the causes of malaria, 30.7% did not take part in malaria control activities. Only 69.3% of the adults regularly sleep inside mosquito nets, 68% of adults buy medicine to cure malaria, 39.9% referred patients to health facilities for cure, and 25% use forest herbs to cure malaria. The factors that increased the malaria morbidity in communes along Vietnam-Lao PDR border have been identified. PMID- 11414470 TI - Expression of cell death-associated proteins in neuronal apoptosis associated with pontosubicular neuron necrosis. AB - Expression of apoptosis-associated proteins p53, bcl-2, bax, and caspase-3/CPP32, activation of caspase-3, and modification of proteins via poly(ADP-ribosyl)ation was studied in pontosubicular neuron necrosis (PSN), a form of perinatal brain damage revealing the morphological hallmarks of neuronal apoptosis. Immunoreactivity for p53 was completely absent. The majority of cells stained with the bax and procaspase-3 antibodies did not show morphological signs of apoptosis. In contrast, an antibody against activated caspase-3 almost exclusively stained cells with apoptotic morphology. Poly(ADP-ribosyl)ated proteins were only rarely detected in cells with apoptotic morphology. The expression patterns of bax, procaspase-3, bcl-2, and p53 in PSN were similar to that found in age-matched control brains. However, activated caspase-3 and poly ADP-ribosylated proteins were exclusively found in apoptotic cells. These data indicate that detection of active caspase-3 is a reliable marker for apoptosis in formalin-fixed human tissue, and that neuronal apoptosis in pontosubicular neuron necrosis is accompanied by a pronounced activation of caspase-3. PMID- 11414471 TI - Transcriptional regulation of caspases in experimental pneumococcal meningitis. AB - Apoptosis and necrosis in brain account for neurological sequelae in survivors of bacterial meningitis. In meningitis, several mechanisms may trigger death pathways leading to activation of transcription factors regulating caspases mRNA synthesis. Therefore, we used a multiprobe RNA protection assay (RPA) to examine the expression of 9 caspase-mRNA in the course of experimental Streptococcus pneumoniae meningitis in mouse brain. Caspase-6, -7 and -11 mRNA were elevated 6 hours after infection. 12 hours after infection caspases-1, -2, -8 and -12 mRNA rose. Caspase-14 mRNA was elevated 18 h and caspase-3 mRNA 24 h after infection. In situ hybridization detected caspases-3, -8, -11 and -12 mRNA in neurons of the hippocampal formation and neocortex. Development of sepsis was paralleled by increased transcription of caspases mRNA in the spleen. In TNFalpha-deficient mice all caspases examined were less upregulated, in TNF-receptor 1/2 knockout mice caspases-1, -2, -7, -11 and -14 mRNA were increased compared to infected control animals. In caspase-1 deficient mice, caspases-11, and -12 mRNA levels did not rise in meningitis indicating the necessity of caspase-1 activating these caspases. Hippocampal formations of newborn mice incubated with heat-inactivated S. pneumoniae R6 showed upregulation of caspase-1, -3, -11 and -12 mRNA. These observations suggest a tightly regulated caspases network at the transcriptional level in addition to the known cascade at the protein level. PMID- 11414472 TI - p73 is not mutated in meningiomas as determined with a functional yeast assay but p73 expression increases with tumor grade. AB - The p53 gene is normally wild type in meningiomas. Since all three members of the p53 gene family recognize the same DNA sequence, tumors containing wild type p53 could decrease transactivation of p53 target genes by mutating either p63 or p73. In meningiomas the most likely target is p73, because loss of heterozygosity of the chromosomal band containing p73 is the commonest genetic lesion in these tumors. To screen p73 for mutations we have developed a functional assay which tests the ability of p73 to activate transcription from a p53-responsive promoter in yeast. The assay correctly identified p73 mutants with mutations equivalent to hotspot mutations in p53, demonstrating that the assay can detect transcriptionally inactive p73. No mutations in p73 were identified in meningiomas. p73 RNA level was higher in more advanced tumors, but there was no correlation between the expression level of p73 and p21, a known p53 target gene. The yeast assay was also used to measure the intrinsic sensitivity of the p73 protein to mutagenesis. Like p53, p73 is exceptionally easy to inactivate as a transcription factor by point mutation. Taken together, these results indicate that p53 and p73 serve very different functions in tumors. PMID- 11414473 TI - Crosstalk between components of the blood brain barrier and cells of the CNS in microglial activation in AIDS. AB - During the progression of AIDS, a majority of patients develop cognitive disorders such as HIV encephalitis (HIVE) and AIDS dementia complex (ADC), which correlate closely with macrophage infiltration into the brain and microglial activation. Microglial activation occurs in response to infection, inflammation and neurological disorders including HIVE, Alzheimer's disease, Parkinson's disease and multiple sclerosis. Microglia can be activated by immunoreactive cells independent of, but enhanced by HIV infection, from at least two routes. Activation may occur from signals originating from activated monocytes and lymphocytes in the blood stream, which initiate a cascade of stimuli that ultimately reach microglia in the brain or from activated macrophages/microglia/astrocytes within the brain. Effects of microglial activation stemming from both systemic and CNS HIV infection act together to commence signaling feedback, leading to HIVE and increased neurodegeneration. Most recent data indicate that in AIDS patients, microglial activation in the brain with subsequent release of excitotoxins, cytokines and chemokines leads to neurodegeneration and cognitive impairment. Since the presence of HIV in the brain results from migration of infected monocytes and lymphocytes across the vascular boundary, the development of novel therapies aimed at protecting the integrity of the blood brain barrier (BBB) upon systemic HIV infection is critical for controlling CNS infection. PMID- 11414474 TI - Clonality of pituitary tumours: more complicated than initially envisaged? AB - The application of allelotype microsatellite polymorphisms and X chromosome inactivation analysis in samples from women allow assessment of clonality. Early studies showed that sporadic human pituitary tumors are benign adenomas of monoclonal origin. This implies that they arise from de novo somatic mutation(s) within a single pituitary cell. However, the evidence obtained from a number of studies indicate that morphology cannot predict clonality, clonality within a given tumour may be multiple or single, multiple tumours arising on the background of hyperplasia may be of identical or differing clonality, and multiple "sporadic" tumours within a gland may be of differing clonal origin. Thus, while the early available evidence indicated that pituitary tumours appear largely monoclonal, it is simplistic to assume that this is inevitable and that these cannot be multiclonal in origin. These observations would be entirely compatible with an initiating stimulus resulting in hyperplasia of specific cell types in the pituitary, which itself gives rise to several distinct clones with variable potential to develop into tumours. Such stimuli might include hypothalamic trophic factors, intrapituitary growth factors, or pituitary specific oncogenes. PMID- 11414475 TI - Oncogene activation in pituitary tumors. AB - Pituitary tumors constitute 10% of intracranial neoplasms and are mostly benign, monoclonal adenomas derived from single mutant cells. Pituitary oncogenes have been intensively studied and three of them, gsp, ccnd1, and PTTG are abundant in significant numbers of cases. gsp is present in approximately 40% of Caucasian patients with GH-secreting tumors and results from a mutated, constitutively active alpha subunit of Gs protein. Persistent activation of the cAMP-PKA-CREB pathway may lead to uncontrolled cell proliferation and GH secretion. ccnd1 is overexpressed cyclin D1, and cyclin D1 gene is amplified in some pituitary tumors. PTTG is expressed in most pituitary tumors. PTTG is localized to both the nucleus and cytoplasm and interacts with several protein partners. At least three tumorigenesis mechanisms are proposed for human PTTG. 1) PTTG and FGF form a positive feedback loop and stimulate tumor vascularity. 2) PTTG transactivates c myc or other pro-proliferation genes. 3) PTTG overexpression causes aneuploidy. PTTG expression activates p53 and causes p53-dependent and -independent apoptosis. Due to lack of functional human pituitary cell cultures and appropriate animal models for pituitary tumors, many of the results reviewed here are obtained from heterologous systems. PMID- 11414476 TI - Tumor suppressor loss in pituitary tumors. AB - The current model of human neoplasia invokes a number of potential genomic alterations that impact cellular phenotype and proliferative rates. In the majority of human tumor models, the transformation from normal cells to neoplastic lesion is a multistep process. This review offers a specific overview of the involvement of tumor suppressor genes (TSGs) in the pathogenesis of human pituitary adenomas. TSG genetic lesions, such as BRCA1 in breast cancer and p53 in Li-Fraumeni Syndrome, have been identified in both sporadic and heritable human endocrine tumors. Familial neoplastic syndromes like multiple endocrine neoplasia type 1 (MEN1) that include pituitary tumor formation as part of a broad clinical spectrum of disease represent a unique opportunity to investigate the general mechanisms of tumorigenesis, and well as genes responsible for sporadic endocrine tumors. Similarly, homologous recombination knockout mice with selectively ablated candidate TSGs have also shed light on the molecular mechanisms of pituitary cell proliferation and tumor suppression. However, despite insights into pituitary tumorigenesis generated by heritable neoplasia syndromes and mouse knockout of critical TSGs that display a pituitary tumor phenotype, the molecular pathogenesis of human pituitary adenomas remains largely an enigma. Thus, the role of TSGs, if any, in sporadic pituitary adenoma formation has yet to be determined, despite our greater understanding of the molecular mechanisms underlying pituitary cell function and phenotype. PMID- 11414477 TI - The role of hormones, growth factors and their receptors in pituitary tumorigenesis. AB - Numerous factors have been shown to govern adenohypophysial cell proliferation. Human and animal models have documented that the hypothalamic trophic hormone growth hormone-releasing hormone stimulates cell proliferation, and prolonged stimulation leads to tumor formation. Similarly, lack of dopaminergic inhibition of lactotrophs and lack of feedback suppression by adrenal, gonadal or thyroid hormones are implicated, perhaps through hypothalamic stimulatory mechanisms, in pituitary adenoma formation superimposed on hyperplasia. However, most pituitary tumors are not associated with underlying hyperplasia. Overexpression of growth factors and their receptors, such as EGF, TGFalpha, EGF-R and VEGF has been identified in pituitary adenomas, and reduction of follistatin expression has been implicated in gonadotroph adenomas. Aberrant expression of members of the FGF family, an FGF antisense gene and FGF receptors have all been described in pituitary adenomas. The clonal composition of pituitary adenomas attests to the molecular basis of pituitary tumorigenesis, however, the evidence suggests that these various hypophysiotropic hormones and growth factors likely play a role as promoters of tumor cell growth in genetically transformed cells. PMID- 11414478 TI - Transgenic and knockout mouse models clarify pituitary development, function and disease. AB - Mouse models have been used to study various aspects of pituitary development, function and disease. Transgenic or knockout technology has been applied to examine the regulation of hormone gene expression and the pathophysiology of its alterations, to ascertain the factors that determine cell differentiation, and to manipulate oncogenesis. Transgenic mice have elucidated the necessary elements required for the tissue- and cell-specific expression of pituitary hormones. Transgenic and knockout technologies have derived mice with hormone overexpression or abrogation of hormone action, and have identified novel hormones. The role of precursor cells in cell differentiation has been confirmed by genetic ablation of cell lineages. Inactivation of transcription factors implicated in pituitary organogenesis and cytogenesis has proven their critical roles in pituitary development. Pituitary oncogenesis has been studied by promoter-directed oncogene expression or tumor suppressor gene ablation, by adenohypophysiotropic hormone overexpression, or by growth factor or receptor overexpression. The tumors have provided a number of cell lines for use in the continuing study of pituitary physiology and pathology. These models may also be used in the future to examine novel therapeutic strategies for the management of patients with pituitary disorders. PMID- 11414479 TI - January 2001: A 37 year old man with a history of Hodgkin's disease. AB - The January Cases of the Month (COM): A case of intracranial metastatic nodular sclerosing Hodgkin's disease without dural attachment in a 37-year-old previously stage III male is presented with a brief review of the literature. Both the primary tumor in the lymph node biopsy and the metastatic brain tumor showed similar histopathology and a immunohistochemical profile typical for Hodgkin's Disease. After chemotherapy, there are no signs of recurrence or systemic disease on follow-up for five months. PMID- 11414480 TI - February 2001: A 74 year old man with a history over 3 months of increasing dyspnea and malaise. AB - The February COM: A man of 78 years with idiopathic late-onset cerebellar ataxia developed renal failure in association with a high ESR and positive pANCA. This was complicated by a subclinical spinal subarachnoid hemorrhage which was related to necrotizing inflammation of small leptomeningeal vessels. Renal cortical infarcts were due to similar inflammation in arcuate and interlobular arteries. Spinal subarachnoid hemorrhage is rare and usually due to rupture of an arteriovenous malformation. However, an immunogenic connective tissue disorder should be considered in the differential diagnosis. In this case, the histology and results of an autoantibody screen support a diagnosis of microscopic polyangiitis. PMID- 11414481 TI - March 2000: A 16 year old female with a cerebellar mass. AB - The March COM: A 16 year old female presented with headaches and cerebellar dysfunction. MR images showed a mass lesion of the right cerebellar hemisphere with mass effect on the medulla. The mass exhibited a striated pattern of alternating isointense and hypointense zones on T1-weighted images that did not contrast enhance. The lesion was hyperintense on T2-weighted images, and also showed a striated appearance. A suboccipital craniotomy and resection of the lesion was performed. Microscopically, the specimen consisted of widened folia and a disorganized cerebellar architectonic pattern in which the internal granular cell layer was occupied by a population of large dysmorphic nerve cell bodies. Patient's diagnosed with Lhermitte-Duclos disease must be adequately evaluated for Cowden's syndrome. PMID- 11414482 TI - Induction of luteolysis in mares by ultrasound-guided intraluteal treatment with PGF2alpha. AB - To evaluate the technique of ultrasound-guided luteal injection in mares, PGF2alpha was administered under ultrasound guidance to horse mares (n = 7 to 9 per group) on Day 9 postovulation via either a systemic (i.m.; zero, 0.01, 0.1, or 5 mg/dose) route or a local intraluteal (i.l.; zero, 0.01 or 0.1 mg/dose) route. The luteolytic efficacy of each treatment was determined based on post treatment decreases in progesterone concentration, interval to uterine edema (IE) and interovulatory interval (IOI). Local administration of PGF2alpha directly into the CL consistently induced luteolysis, at doses up to 50-fold lower than the lowest effective systemic dose. Significant decreases in IOI and IE occurred in mares treated with 5 mg PGF2alpha i.m. or 0.1 mg PGF2alpha i.l., but did not occur in mares treated with 0.1 or 0.01 mg PGF2alpha i.m., 0.01 mg PGF i.l., vehicle i.l. or vehicle i.m.. Progesterone concentrations were reduced to less than 10% of pretreatment values by two days post treatment in mares treated with 5 mg PGF2alpha i.m. or 0.1 mg PGF2alpha i.l.. PGF2alpha doses of 0.1 mg i.m. and 0.01 mg i.l. were associated with smaller but significant progesterone decreases (to 66% and 46% of pre-treatment values, respectively) by two days post treatment. Progesterone values after administration of i.l. vehicle did not differ from pre-treatment values by two days post treatment, but were significantly lower (53% of pre-treatment values) by four days post treatment. Intramuscular treatment with vehicle or 0.01 mg of PGF2alpha did not significantly reduce progesterone concentrations below pretreatment values. Overall, the minimum effective luteolytic dose of PGF2alpha given intraluteally was between 0.01 and 0.1 mg. Based on the results of this study, ultrasound guided i.l. injection appears to be a repeatable method for studying the direct effect of other chemicals on luteal function. However, the current procedure carries some risk, since three i.l. injections were associated with ovarian abscesses. PMID- 11414483 TI - Exogenous growth hormone improves the number of transferable embryos in superovulated ewes. AB - The application of pGH (porcine Growth Hormone) to superovulated ewes was studied with the aim of improving the embryo yield. Thirty-seven ewes were superovulated with pFSH for 3 d and 18 of them were cotreated the third day with 0.50 mg of pGH. Embryos were surgically recovered on Day 7 after sponge withdrawal. Then, 102 morphologically healthy embryos were immediately transferred in pairs to 51 synchronized recipient ewes. The GH treatment did not significantly affect the percentage of ewes in estrus, the time of estrus onset or the ovulation rate. However, it improved synchronization by grouping estrus in a narrower range (12 h) in comparison to the control group (24 h); (16 to 28 h after sponge withdrawal vs 12 to 36 h; P < 0.05). The total amount of LH released during the preovulatory surge was lower in the GH than in the control group (P < 0.05). No differences were found between groups for other LH-related parameters such as basal levels, peak values or peak time from sponge removal. The proportions of unfertilized oocytes and degenerate embryos recovered were lower in the GH cotreated group (P < 0.05 and P < 0.01, respectively). This resulted in higher rates of transferable embryos and lambs born per donor ewe in the GH than in the untreated group (3.9 vs 1.7 and 2.28 vs 0.84, respectively; both, P < 0.05). These beneficial effects of GH would likely be due either to a direct action on oocyte maturation or to an indirect action on the oviductal environment. PMID- 11414484 TI - Estrus and pregnancy after synchrony with lutalyse in conjunction with Syncro Mate-B. AB - Estrous response and pregnancy rates are decreased for cows given Syncro-Mate-B (SMB) during metestrus (Day 1 to 5 of an estrous cycle). Data indicate these decreases are due, in part, to retention of a functional corpus luteum (CL). Our objective was to determine whether PGF2alpha administered in conjunction with SMB would improve estrous response and pregnancy rates in metestrous cows with no detrimental effects to cows in other stages of the estrous cycle. Three hundred seventy-three suckled beef cows were observed for estrus for 21 d before SMB administration to determine stage of an estrous cycle. Blood samples were collected 14 and 7 d before treatment and at SMB administration. Serum was assayed for concentration of progesterone to verify stage of estrous cycle or noncyclicity. All cows received the standard SMB regime and were allotted by age and stage of cycle to one of two groups. Cows denoted SMB + L received 25 mg of PGF2alpha 8 d after implantation, whereas cows denoted SMB served as controls. On Day 10, SMB implants were removed and females were observed for subsequent estrus. At this time, calves were removed from their dams for 48 h. Artificial insemination was performed 12 hr after observation of a standing estrus. Timed insemination was performed at 48 hr after implant removal for cows not inseminated at 24 or 36 hr after implant removal. Interval to synchronized estrus (within 5 d of implant removal) was lengthened for metestrous cows compared to cows in other stages of the cycle irrespective of treatment (P < 0.001). Cows receiving PGF2alpha had a greater pregnancy rate at 5 d compared to controls (P = .0672). Interval to estrus, estrous response, and pregnancy rate to A1 at d 28 or end of breeding season were not affected by administration of PGF2alpha in conjunction with SMB when compared to the standard SMB protocol. PMID- 11414485 TI - Ram-induced ovulation to improve artificial insemination efficiency with frozen semen in sheep. AB - Ram effect, defined as shortening of seasonal anestrus in ewes by exposure to the ram, is now well recognized but the underlying mechanisms are still unclear. Little information also exists whether the ram is able to influence the estrus cycle and ovulation. Three experiments were conducted to investigate endocrine response, time of ovulation and pregnancy rate of ewes in proestrus, exposed to the ram (treated) or an adult ewe (control). In the first experiment, ewes (n = 20) were treated with fluorgestone acetate pessaries for 12 days and were given eCG and cloprostenol one day before withdrawal of pessaries. On the day after removal of the pessaries ewes in the treated group (n = 10) were exposed to the ram and those in the control group (n = 10) were exposed to an adult ewe. Blood samples were taken for LH assay every 20 min from 2 h before to 24 h after ram exposure. In the second experiment, ewes (n = 120) were induced into proestrus and on the day after removal of the pessaries were exposed to either a ram (n = 60) or a ewe (n = 60) as described above and were laparoscoped 50, 60 or 70 h after pessary withdrawal (n = 20 at each time interval). In the third experiment ewes (n = 90) were induced and exposed to the ram (n = 45) or an adult ewe (n = 45) and inseminated via a laparoscope whit frozen-thawed semen at 50 or 60 h after pessary removal, respectively. Exposure to the ram was followed in 2 h by a marked rise in LH, equivalent to a preovulatory surge in duration and amplitude. It was also followed by concentrated ovulation within 25 to 30 h and by an increased pregnancy rate in exposed ewes (73.3 vs. 53.3%). PMID- 11414486 TI - The effect of estradiol benzoate on synchrony of estrus and fertility in cattle after removal of a progesterone-releasing intravaginal device. AB - The aim was to determine the effect of estradiol benzoate (EDB) given after removal of a progesterone-releasing intravaginal device (PRID) at either emergence or dominance of a follicle wave, on the interval to estrus, variation in its onset and pregnancy rate in heifers. Heifers (n=186) were assigned randomly to four treatments in a 2 x 2 factorial design; emergence or dominance of a follicle wave at PRID removal, with or without 0.5 mg EDB 24 h after PRID removal. Ovarian ultrasonography was performed to confirm follicular status; data from heifers of undeterminable follicular status were excluded (n=36). Mean size of the largest follicle of the new wave at PRID removal was smaller (P < 0.01) in heifers given EDB at emergence (6.3 +/- 0.09 mm) compared with those given it at dominance (10.9 +/- 0.30 mm). The onset of estrus was earlier (P < 0.01) in heifers given EDB at dominance (median 42 h, range 13 h) compared with those not given EDB at dominance (median 43 h, range 42 h). The median interval to estrus was decreased (P < 0.01) in heifers given EDB at emergence (median 48 h, range 73 h) compared with those not given EDB at emergence (median 66 h, range 45 h). Variation in onset of estrus was reduced (P < 0.05) in heifers given EDB compared with those not given EDB. The pregnancy rate was not affected when EDB was given at dominance, however, it was decreased (P < 0.05) when given at emergence (23 of 40 vs 26 of 32, respectively). To determine the effect of EDB on follicular dynamics in heifers treated with EDB at emergence, heifers (n=37) were assigned to two treatments: at emergence with or without EDB and their ovaries were examined daily using ultrasonography. Follicular dynamics were not different (P > 0.05) in EDB-heifers compared with untreated controls. Mean serum estradiol was greater (P < 0.01) in EDB-treated heifers compared with controls. In conclusion, 0.5 mg EDB given 24 h after PRID removal to heifers decreased the interval to estrous onset at emergence or dominance, decreased variation in onset of estrus and decreased pregnancy rates when given at emergence of a follicle wave. PMID- 11414487 TI - Effects of semen filtration and dilution rate on morphology and fertility of frozen gander spermatozoa. AB - Feces, urates or dirt originating from feathers often contaminate gander semen during collection, threatening its fertilizing ability. Seminal plasma used as a diluent has a similar effect, particularly on spermatozoa subjected to cryopreservation or short-term storage under refrigeration. The aim of the experiments was to evaluate the effects on spermatozoa motility, morphology and fertilizing ability after minimizing the influence of the contaminants by semen filtration or dilution prior to freezing. Pooled semen, collected twice a week from 9 White Italian ganders by dorso-abdominal massage, was divided into two parts. One sample was filtered and both were diluted in 1:1 or 1:0.5 (v/v) with EK diluent, equilibrated for 15 min at +4 degrees C, mixed with dimethyl acetamide (DMA) in the final concentration 6% (v/v) and frozen to -140 degrees C in a computerized freezer, at a rate of 60 degrees C/min. In fresh and processed (filtered, freeze-thawed) semen were examined the spermatozoa motility and morphology, and fertilizing ability for freeze-thawed semen, both for unfiltered and filtered. In freeze-thawed semen no tangible differences due to experimental factors were observed in motility and percent of live spermatozoa in total. On average 35 to 42% of the spermatozoa survived the freezing process, but only 10 to 15% were normal, without any damage visible under the light microscope. The fertility of unfiltered freeze-thawed semen inseminated twice a week in a 0.2 mL dose (about 3 to 5 x 10(6) of live normal spermatozoa each) averaged 66.1% and hatchability of the set eggs 57.1 and 86.5% of the fertile eggs. The fertility obtained after the insemination with semen filtered prior to freezing was lower (64.3%), but hatchability was slightly higher (58.6 and 91.1% of set and fertile eggs, respectively). The duration of fertility for filtered semen was longer than that for unfiltered, 10 days after the last insemination the eggs were still fertile. The fertility results of freeze-thawed gander semen were very promising taking into consideration the small amount of inseminated live normal spermatozoa and it is possible to improve this result by increasing the number of spermatozoa in the insemination dose. PMID- 11414488 TI - Effect of short-term treatment with bovine somatotropin at estrus on conception rate and luteal function of repeat-breeding dairy cows. AB - We studied the effect of recombinant bovine somatotropin (rbST) at the time of estrus on progesterone concentrations and conception rates of repeat-breeding Holstein cows. We used repeat-breeding cows of varied parity (n = 510). All the animals were clinically healthy and had had at least three unsuccessful services before entering the study. After detection of estrus, the cows were randomly assigned to either a treated (n = 201) or a control (n = 309) group. The animals in the treated group were given rbST (500 mg s.c.) at the time of estrus and again 10 d later. Artificial insemination was performed 12 h after the first detection of estrus. In order to evaluate the effect of rbST on luteal function, blood samples were taken from 10 cows in each group every 3 d for 18 d, starting on the day of insemination (Day 0) to determine progesterone concentrations. Conception rates were significantly higher (P < 0.05) in the cows treated with rbST (29.3%) than in the control cows (16.9%). The effects of rbST were maximal in cows with 8 or more previous unsuccessful services and in cows with 2 to 4 calvings. Progesterone concentrations tended to be higher in nonpregnant cows that were treated with rbST than in those that were not treated. The difference between groups was significant (p < 0.05) on Day 18 after insemination. In pregnant cows there were no significant differences in progesterone concentrations between treated and nontreated animals at any time. Treatment with rbST at estrus improved the conception rate of repeat-breeding Holstein cows. This effect was associated with an increase in circulating progesterone concentrations on Day 18. PMID- 11414489 TI - Rapid sexing of preimplantation bovine embryo using consecutive and multiplex polymerase chain reaction (PCR) with biopsied single blastomere. AB - The objective of this study was to establish a rapid and reliable PCR method for the sexing of 8- to 16-cell stage bovine embryos. The BOV97M and bovine 1.715 satellite DNA sequences were selected for amplification of male- and bovine specific DNA, respectively. But the unequal number of copies of these two repetitive sequences required some modification of the multiplex PCR method. In consecutive and multiplex PCR, the first 10 PCR cycles were done with male specific primer followed by an additional 23 cycles with bovine-specific primer. In this PCR method, the appearance of male- and bovine-specific bands was independent of the DNA concentration. This PCR method was applied successfully using groups of 8, 4, 2, and 1 blastomeres dissociated from the embryos, and the sexing efficiency was 100.0, 96.3, 94.3 and 92.1%, respectively. The coincident rate of sex determination between biopsied single blastomere and matched blastocyst was 90.0%. Therefore the developmental potential from 8- to 16-cell stage embryos to the blastocyst stage was not significantly different (P>0.2) for intact embryo (42.3%) than for demi-embryos (53.8%), suggesting that trauma to the demi-embryo caused by single-blastomere aspiration using a bevelled micropipette was very small. In conclusion, we developed a rapid (within 2 hours) and effective PCR method for the sexing of 8- to 16-cell stage bovine embryos using a single blastomere. PMID- 11414490 TI - Fertilization and embryo recovery rates in superovulated chios ewes after laparoscopic intrauterine insemination. AB - Forty superovulated dairy ewes of the Greek Chios breed were used in an experiment to evaluate the efficiency of laparoscopic intrauterine insemination on fertilization and embryo recovery rates as well as embryo quality. Estrus was synchronized by intravaginal progestagen impregnated sponges and superovulation was induced by administration of 8.8 mg o-FSH i.m. following a standard 8 dose protocol. A small volume (0.3 mL) of diluted fresh ram semen was deposited in each uterine horn 24 to 28 h after onset of the estrus by a laparoscopic technique. The animals were allocated randomly into two groups (Group A and B) of 20 animals each. In Group A, embryos were recovered 18 to 24 h after the intrauterine insemination and in Group B on Day 6. The average number of corpora lutea was 12.8 +/- 1.2 and 11.5 +/- 1.1 (+/- SEM); the overall embryo recovery was 66.4% and 57% and the percentage of recovered fertilized ova was 81% and 82.8% in Groups A and B, respectively. More fertilized ova were collected per ewe from Group A (P < or = 0.1). Results indicated that in Chios breed, superovulation using homologous FSH combined with laparoscopic AI leads to good ovarian response with satisfactory results in fertilization, embryo recovery and quality of embryos. This could lead to improved and more efficient methods for obtaining large numbers of high quality oocytes and embryos for embryo transfer programs which could contribute to genetic improvement and increase of the population size. PMID- 11414491 TI - Isolation and purification of the ovulation-inducing factor from seminal plasma in the bactrian camel (Camelus bactrianus). AB - The purpose of this study was to extract, identify and partially characterize a newly found ovulation-inducing factor, and thus gain our understanding of induced ovulation in biology. In our preliminary research, an ovulation-inducing factor (OIF) was isolated and purified from seminal plasma of the bactrian camel by ion exchange chromatography on DEAE-cellulose, HPLC and reverse-phase HPLC. The OIF is a peptide with 74 residues and GnRH-like bioactivity, which is heat-stable in camel seminal plasma because the OIF is wrapped up in several protein layers with different properties. However, purified OIF is degraded when it is exposed to oxygen or is heated in water. According to the analysis of amino acid components and partial amino acid sequence aminated N-terminus, and its molecular weight, the OIF is completely different from the native-LHRH, LH, HCG, PMSG and PGF 2alpha. The OIF is a novel ovulation hormone in the bactrian camel, but is similar to that reported in the bull. PMID- 11414492 TI - Intrabursal transfer of spermatozoa (ITS): a new route for artificial insemination of mice. AB - Artificial insemination (AI) by direct injection of epididymal spermatozoa into the reproductive tract of females is simpler and more convenient than in vitro fertilization (IVF) and subsequent transfer of fertilized eggs to recipient oviducts for simultaneous acquisition of a large number of pups. Introduction of epididymal spermatozoa into oviducts via the oviductal wall or via vaginal and intrauterine routes is currently the most commonly used method for AI in mice. In this study, we explored another route for AI of the mouse and found that transfer of spermatozoa into a space near the infundibulum between the ovary and ovarian bursa enables in vivo fertilization of ovulated oocytes at the ampulla. When 1 microL of a sperm suspension containing 1 x 10(4) spermatozoa freshly isolated from B6C3F1 males was intrabursally injected into superovulated B6C3F1 females on E (embryonic day) 0.4 (10:00 AM), 5 of 7 females yielded 2-cell embryos with rates of efficiency ranging from 4 to 21% (11% on average), which were much lower than those (91% on average) for embryos obtained by natural mating. All the 2 cell embryos derived from injection of sperm developed in vitro to hatched blastocysts. Similar results were obtained from injection of 1 microL of sperm suspension containing 1 x 10(3) spermatozoa, although in vivo fertilizing ability was slightly improved (28% on average). When 1 microL of sperm suspension containing 1 x 10(4) spermatozoa was injected intrabursally into superovulated females that had been mated with vasectomized males, 6 of 10 mice (60%) yielded 19 normal mid-gestational fetuses with an average litter size of 3.2, which was much lower than that (14.5) for embryos obtained by natural mating. Although the present findings appear to be preliminary, this technique, based on the intrabursal transfer of spermatozoa, will be of practical use for AI in mice, particularly for transgenic and mutant mice that are often difficult to breed. PMID- 11414493 TI - Effect of periovulatory prostaglandin F2alpha on pregnancy rates and luteal function in the mare. AB - The objective of this study was to determine whether periovulatory treatments with PGF2alpha affects the development of the CL, and whether the treatment was detrimental to the establishment of pregnancy. Reproductively sound mares were assigned randomly to one of the following treatment groups during consecutive estrus cycles: 1. 3,000 IU hCG within 24 hours before artificial insemination and 500 microg cloprostenol (PGF2alpha analogue) on Days 0, 1, and 2 after ovulation (n=8), 2. 2 mL sterile water injection within 24 hours before artificial insemination and 500 microg cloprostenol on Days 0, 1, and 2 after ovulation (n=8); 3. 3,000 IU hCG within 24 hours before artificial insemination and 500 microg cloprostenol on Day 2 after ovulation (n=8); or 4. 3,000 IU hCG within 24 hours before artificial insemination and 2 mL of sterile water on Days 0, 1, and 2 after ovulation (controls; n=8). Blood samples were collected from the jugular vein on Days 0, 1, 2, 5, 8, 11, and 14 after ovulation. Plasma progesterone concentrations were determined by the use of a solid phase 125I radioimmunoassay. All mares were examined for pregnancy by the use of transrectal ultrasonography at 14 days after ovulation. Mares in Group 1 and 2 had lower plasma progesterone concentrations at Day 2 and 5, compared to mares in the control group (P < 0.001). No difference was detected between group 1 and 2. Plasma progesterone concentrations in group 3 were similar to the control group until the day of treatment, but decreased after treatment and were significantly lower than the control group at Day 5 (P < 0.001). Plasma progesterone concentrations increased in all treatment groups after Day 5, and were comparable among all groups at Day 14 after ovulation. Cloprostenol treatment had a significant effect on pregnancy rates (P < 0.01). The pregnancy rate was 12.5% in Group 1, 25% in Group 2, 38% in Group 3, and 62.5% in Group 4. It was concluded that periovulatory treatment with PGF2alpha has a detrimental effect on early luteal function and pregnancy. PMID- 11414494 TI - Prostaglandin F2alpha added to extended boar semen at processing elicits in vitro myometrial contractility after 72 hours of storage. AB - Improved fertility will maximize productivity of the swine industry. Myometrial contractility is an essential component in the fertilization process because it is the mechanism by which spermatozoa are transported to the site of fertilization. In the present study, we evaluated the potential use of PGF2alpha supplementation to the extended pig semen in regard to inducing myometrial contractility of sows. Extended boar semen (80 mL) was supplemented with PGF2alpha (5 mg) for 72 h at 17 +/- 1 degrees C. Cumulative doses of 0.1, 1, 10 and 100 microL of the mixture were tested on uterine strips obtained from diestrus sows. An increase in myometrial contractility was recorded with PGF2alpha supplementation when compared to extended semen or extender treatment alone after 72 h of incubation. Addition of PGF2alpha to the extended boar semen at the time of the experiment did not differ from the 72 h treated group. The results from this study support that PGF2alpha preparations can be added to extended doses of boar semen at processing to enhance myometrial contractility at the time of insemination for up to 72 h. PMID- 11414495 TI - Amino acid concentrations in fluids from the bovine oviduct and uterus and in KSOM-based culture media. AB - Amino acids in bovine oviductal and uterine fluids were measured and compared with those in modified simplex optimized medium (KSOM) supplemented with either fetal calf serum or Minimum Essential Medium amino acids in addition to bovine serum albumin, fetal calf serum or polyvinyl alcohol. Concentrations of cysteine, threonine, tryptophan, alanine, aspartate, glycine, glutamate, proline, beta alanine, and citrulline were higher in oviductal fluids than in KSOM-based culture media. Nonessential and essential amino acids were present in ratios of 5:1 and 2:1 in oviductal and uterine fluids, respectively. Concentrations of alanine (3.7 mM), glycine (14.1 mM) and glutamate (5.5 mM) were high in oviductal fluids, comprising 73% of the free amino acid pool. Of the amino acids measured in uterine fluids, alanine (3.1 mM), glycine (12.0 mM), glutamate (4.2 mM), and serine (2.7 mM) were highest in concentration, and the first three comprised 43% of the free amino acid pool. In conclusion, amino acid concentrations in the bovine reproductive tract were substantially higher than those in embryo culture media. Certain amino acids, particularly alanine, glutamate, glycine and taurine, are present in strikingly high concentrations in both oviductal and uterine fluids, suggesting that they might play important roles in early embryo development. The particular pattern of amino acid concentrations may be an important factor to be considered for the improvement of embryo culture media. PMID- 11414496 TI - Restoring ovulation in beef donor cows with ovarian cysts by progesterone releasing intravaginal silastic devices. AB - The objective of this study was to determine the efficacy of a progesterone releasing intravaginal silastic device (Controlled Internal Drug Release: CIDR) for inducing ovulation in beef cows with persistent ovarian cysts. Fifteen cows with cysts and abnormal cycles for over 40 days were randomly assigned to receive either a single CIDR (CIDR group, n=9), or a CIDR containing no progesterone (blank CIDR) (BLANK group, n=6) for about 14 days. Determination of plasma progesterone levels at the beginning of CIDR treatment indicated 4 of 6 BLANK cows with non-luteinized cysts and 5 of 9 CIDR cows with non-luteinized cysts. In 5 of 6 BLANK cows, one follicular wave appeared and newly emerged dominant follicles increased in size up to 20 mm in diameter and persisted during the experiment, while one cow experienced estrus with spontaneous ovulation. In contrast, during CIDR treatment, 2 or 3 waves, in which dominant follicles were from 7 to 15 mm in diameter, appeared approximately at 7-day intervals. Within 3 days after CIDR removal, estrous behavior was detected followed by ovulation of the dominant follicle in the last wave. All CIDR cows resumed normal cyclicity with 2 follicular waves for over 2 months. Insertion of a CIDR caused a rapid increase of about 2 ng/mL in plasma progesterone. The levels were greater than 1.3 ng/mL until removal of a CIDR, then dropped under 0.3 ng/mL. Concentrations of plasma estradiol in BLANK cows increased during growth of the cystic follicles, with high levels greater than 10 pg/mL for over 10 days. In 4 of 5 cows with non-luteinized cysts, with high plasma estradiol on the day of CIDR insertion, CIDR treatment resulted in rapid decline of estradiol levels. During placement of the CIDR, estradiol levels showed no increase in the growth phase of a newly appeared dominant follicle. After CIDR removal, however, estradiol significantly increased associated with the growth of ovulatory follicles in all 9 cows. A transient increase in plasma FSH levels preceded detection of each follicular or cyst wave in both BLANK and CIDR cows. Pulse frequency and mean concentration of LH in cows with non-luteinized cysts showed values corresponding to those in normal follicular phase. However, throughout CIDR treatment, these parameters reduced to levels found in the normal luteal phase. In cows with luteinized cysts, parameters of LH secretion were as low as in the normal luteal phase before and during CIDR treatment, then increased significantly after CIDR removal. Present results indicate that treatment with CIDR proved effective in restoring ovulation and reestablishing normal cyclicity in beef donor cows with cysts persistent for a long period. The CIDR reduced and maintained LH secretion at normal luteal levels, thereby, inducing atresia of estrogen-active cysts and preventing formation of cysts from the newly emerged follicles. PMID- 11414497 TI - Development of goat embryos after in vitro fertilization and parthenogenetic activation by different methods. AB - Effective activation protocols that can be used during nuclear transfer investigations in goats need to be developed. We compared the development of IVF goat embryos with those of nonfertilized parthogenetically developing oocytes activated by treatment with either ionomycin or ethanol, both followed by immediate exposure to 6-diethylaminopurine (6-DMAP). Cumulus oocyte complexes (COCs) recovered from abattoir goat ovaries were either matured in a conventional laboratory incubator or placed in pre-equilibrated maturation medium and shipped overnight in a battery-operated dry incubator to another laboratory. Mature COCs were allocated randomly to one of three treatment groups. Group 1 oocytes (n=169 shipped, n=253 not shipped) were fertilized in vitro at 24 h postmaturation (hpm). The remaining COCs were activated at 28 hpm in either ionomycin (Group 2: n=362 shipped, n=202 not shipped), or ethanol (Group 3: n=263 shipped, n=249 not shipped). Activated oocytes were immediately incubated in 6-DMAP for 4 h. Blastocyst development was evaluated on Day 8 post-insemination/activation. Percent cleavage was comparable in shipped and nonshipped oocytes and in all treatment groups. In both shipped and nonshipped oocytes, parthenotes developing from ionomycin- and ethanol-activated oocytes had significantly greater blastocyst development (P<0.01) compared to IVF embryos (28.5 +/- 3.0, 27.4 +/- 2.8, 10.3 +/- 3.0, respectively for the nonshipped oocytes and 9.9 +/- 2.1, 10.3 +/- 2.4, 3.7 +/- 4.7 respectively for the shipped oocytes). Shipped oocytes had lower blastocyst development compared to nonshipped oocytes in the three treatment groups. The mean blastocyst cell number was not statistically different between shipped and nonshipped oocytes or among treatment groups, suggesting that all were equally viable. PMID- 11414498 TI - Effect of culture medium pH on bacterial gellan production. AB - The effect of the initial pH of the culture medium used in the production of the exopolysaccharide gellan by the bacterium Pseudomonas species ATCC 31461, when glucose or corn syrup served as the carbon source, was investigated. With glucose as the carbon source, exopolysaccharide formation was highest after 72 h of growth when the initial pH of the culture medium was 6.8 to 7.4. Polysaccharide production by the bacterial cells grown on corn syrup for 72 h was maximal when the initial pH of the medium was 7.0 or 7.2. Cell weights of the strain after 72 h tended to be higher for the glucose-grown cells than for the corn syrup-grown cells. PMID- 11414499 TI - Identification of Thiobacillus ferrooxidans strains based on restriction fragment length polymorphism analysis of 16S rDNA. AB - The 16S rDNA sequences from ten strains of Thiobacillus ferrooxidans were amplified by PCR. The products were compared by performing restriction fragment length polymorphism (RFLP) analysis with restriction endonucleases Alu I, Hap II, Hha I, and Hae III. The RFLP patterns revealed that T. ferrooxidans could be distinguished from other iron- or sulphur-oxidizing bacteria such as T. thiooxidans NB1-3, T. caldus GO-1, Leptospirillum ferrooxidans and the marine iron-oxidizing bacterium strain KU2-11. The RFLP patterns obtained with Alu I, Hap II, and Hae III were the same for nine strains of T. ferrooxidans except for strain ATCC 13661. The RFLP patterns for strains NASF-1 and ATCC 13661 with Hha I were distinct from those for other T. ferrooxidans strains. The 16S rDNA sequence of T. ferrooxidans NASF-1 possessed an additional restriction site for Hha I. These results show that iron-oxidizing bacteria isolated from natural environments were rapidly identified as T. ferrooxidans by the method combining RFLP analysis with physiological analysis. PMID- 11414500 TI - A new extraction method for Acinetobacter species ODB-L2 rough form lipopolysaccharide from culture broth. AB - The Gram-negative bacterium Acinetobacter species ODB-L2 produces lipopolysaccharide (LPS) in culture broth. The LPS could not be purified by conventional extraction methods using 90% phenol/water or 90% phenol/chloroform/petroleum ether mixed solvent. Extraction was achieved employing an admixture of chloroform, ethanol, and 4 M HCI solution. The LPS was purified from dissolving the crude extracts in 90% phenol and LPS sediment formed by addition of methanol. The LPS was characterized by chemical, biochemical, and physicochemical methods as rough form 3-hydroxydodecanoic acid rich LPS. PMID- 11414501 TI - The effects of subminimal inhibitory concentrations of beta-lactam antibiotics against Clostridium perfringens. AB - The effects of subminimal inhibitory concentrations (sub-MIC) of four beta-lactam antibiotics [penicillin-G (PCG), ampicillin (AMP), cephaloridine (CER), cephalothin (CET)] were tested against Clostridium perfringens type A PB6K, after determining the minimum inhibitory concentrations (MIC) of 29 different Clostridium strains. The majority of the strains were sensitive to all beta lactam antibiotics. Morphological changes, such as filamentous development and lysis, occurred at concentrations considerably lower than the MIC of CER and CET in C. perfringens. Clear cooperation of AMP and CER with rabbit polymorphonuclear leucocytes (PMNL) against C. perfringens was observed. The filamentous bacteria produced as a result of exposure to sub-MIC of each antibiotic, were phagocytosed easily. The ratios between the drug concentrations (microg/ml) at which the morphological changes began to occur, the minimum antibiotic concentrations (MAC), and the MIC values (microg/ml), were calculated. A large ratio indicated a wide range of effective concentrations below the MIC value for the antibiotics. PMID- 11414502 TI - Contamination and survival of Pseudomonas aeruginosa in hospital used sponges. AB - The microbial contamination of in-use sponges was investigated. Of the sixty sponges examined, 51 (85%) were contaminated with 10(3)-10(9) colony forming units (CFU) per sponge. Pseudomonas aeruginosa was isolated from sixteen (26.7%) of the sixty sponges. P. aeruginosa survived for 2 months in contaminated sponges which were left at room temperature and became dry to the touch. The susceptibility of sponges to P. aeruginosa contamination should be recognized. Once sponges are contaminated with P. aeruginosa, eradication of this organism is difficult even if the sponges are dried. PMID- 11414503 TI - Antibacterial activity of Karanj (Pongamia pinnata) and Neem (Azadirachta indica) seed oil: a preliminary report. AB - The antibacterial activity of Karanj (Pongamia pinnata) and Neem (Azadirachta indica) seed oil in vitro against fourteen strains of pathogenic bacteria was assessed. Using the tube dilution technique, it was observed that 57.14 and 21.42% of the pathogens were inhibited at 500 microl/ml; 14.28 and 71.42% at 125 microl/ml; and 28.57 and 7.14% at 250 microl/ml of Karanj and Neem oils, respectively. The activity with both the oils was bactericidal and independent of temperature and energy. Most of the pathogens were killed more rapidly at 4 degrees C than 37 degrees C. The activity was mainly due to the inhibition of cell-membrane synthesis in the bacteria. PMID- 11414504 TI - Prostaglandylinositol cyclic phosphate synthase activity in the liver of insulin resistant rhesus monkeys before and after a euglycemic hyperinsulinemic clamp. AB - Prostaglandylinositol cyclic phosphate (cPIP), functionally a cAMP antagonist, is a novel, low-molecular weight mediator of insulin action. Both essential hypertension and type 2 diabetes may be associated with a reduction of cPIP synthesis. In intact cells and in plasma membranes, cPIP synthesis is stimulated by insulin, which activates cPIP synthase by tyrosine phosphorylation. We measured the activities of cPIP synthase in the homogenates of freeze-clamped and then lyophilized liver samples from five insulin-resistant, adult rhesus monkeys, obtained under basal fasting conditions and again under maximal insulin stimulation during a euglycemic hyperinsulinemic clamp. The mean cPIP synthase activity in basal samples (0.33 +/- 0.09 pmol/min/mg protein) was not significantly different at the end of the clamp (0.24 +/- 0.11 pmol/min/mg protein). Basal cPIP synthase activityVoL 12, No. 1, 2001 was directly related to both basal cAMP content and basal fractional activity of cAMP-dependent protein kinase (PKA): r=0.85, p<0.05 and r=0.86, p<0.05, respectively. In turn, insulin stimulated cPIP synthase activity was inversely related to both the insulin stimulated fractional activity of PKA (r=0.89, p<0.02) and the insulin-stimulated total PKA activity: r=0.94, p<0.005. The findings suggest that in the liver of insulin-resistant rhesus monkeys, cPIP synthase activity, which leads to the synthesis of the low-molecular weight mediator cPIP, may oppose cAMP synthesis and PKA activity. PMID- 11414505 TI - Changes in heart rate variability following acclimation to heat. AB - We studied the sympatho-vagal balance during acclimation to heat in eight healthy individuals. The subjects, dressed in shorts and tennis shoes, underwent a 10 d procedure of acclimation. Daily exposure lasted 115 min--5 min rest followed by 2 bouts of 50 min exercise (walking on a treadmill at a work load of approximately 350 watt) separated by 10 min rest-at 40 degrees C and 40% relative humidity. We analyzed the time and frequency domains of 256 R-R intervals, toward the end of the second bout of exercise, on the first and tenth days of acclimation. Typical changes for heat acclimation (a reduction in rectal temperature and heart rate, an increase in sweat rate) were observed. Concomitant to a decrease in the final heart rate on the tenth day, sympathetic tone increased. We suggest that, except for the chronotropic response, changes in the sympatho-vagal balance may reflect several adaptive mechanisms of the cardiovascular system. Peripheral factors rather than intrinsic cardiac factors are of relatively greater importance in the direct control of cardiac function. PMID- 11414506 TI - Increased cardiac alpha-1-adrenoceptor density in rats following treatment with amiodarone. AB - This study was undertaken to investigate the interaction between amiodarone and alpha-1-adrenoceptors in rat cardiac cells. The level (Bmax) and affinity (Kd) of alpha-1-adrenoceptors in heart cells were determined by [3H]prazosin radioligand binding following amiodarone treatment. In cultured intact cardiocytes treated for 48 h with 10 microM amiodarone, [3H]prazosin binding increased by 31% compared with the control cells (p<0.05). The increase was both dose and time dependent and was found to be specific because no significant change occurred in creatine kinase activity. Additionally, under the same conditions, an increase in [3H]prazosin binding to cultured cardiocyte cell membranes was also obtained. Oral gavage of amiodarone to rats for 8 d resulted in a 25% increase in [3H]prazosin binding to isolated ventricle membranes compared with control rats (p<0.05). We conclude that amiodarone treatment can increase the response to alpha-1-adrenoceptors agonist in the heart due to an increase in the density of alpha-1-adrenoceptors. PMID- 11414507 TI - Responses to animated contours of neurons in visual cortex area 18 of the cat. AB - Given that cells in visual cortical area 18 in cats encode information of complex shapes, in addition to conventional stimuli like bars and gratings, we studied the capability of single cells in this area to 'identify' animated contours. The recorded cells were not selective to specific animated contours, whereas they were highly sensitive to the contour's degradation and to the orientation of its elements. The results indicate that cells in area 18 can encode information about many characteristics of complex animated patterns. The findings suggest the existence of a contour abstracting mechanism, which needs further study. PMID- 11414508 TI - Lipid peroxidation and glutathione peroxidase activity in chronic obstructive pulmonary disease exacerbation: prognostic value of malondialdehyde. AB - Results of recent studies have indicated that during exacerbation of chronic obstructive pulmonary disease (COPD), antioxidant capacity is lower and the levels of lipid peroxidation products are higher than those in age-matched healthy subjects. The aim of this study was to assess the time course of changes in oxidant stress during the treatment of exacerbation of COPD. For this purpose, we measured erythrocyte glutathione peroxidase (GPx) activity and serum levels of the lipid peroxidation product malondialdehyde (MDA) in 18 male patients with acute exacerbation of COPD. Fifteen healthy non-smokers having no history of lung disease served as control subjects. Mean erythrocyte GPx values of patients were 45.54 +/- 9.04 u/gHb on admission and had increased to 72.77 +/- 9.68 by the tenth day of treatment, but still remained lower than those of healthy subjects (83.13 +/- 10.91) (p=0.007). Serum MDA values in patients were Vol. 12, No. 1, 2001 significantly higher (2.68 +/- 1.28 nmol/ml) than those in control subjects (1.04 +/- 0.36 nmol/ml) (p=0.000) and returned to normal values by the tenth day of treatment (1.08 +/- 0.36 nmol/ml) (p=0.766). Erythrocyte GPx values in patients who were current smokers (39.87 +/- 3.82 u/gHb) were lower than those in ex-smokers (49.15 +/- 9.67 u/gHb) (p=0.021). Moreover, serum MDA values in patients who were current smokers (3.32 +/- 1.18 nmol/ml) were higher than those in ex-smokers (1.66 +/- 0.60 nmol/ml) (p=0.007). The results show that oxidative stress in patients with acute exacerbation of COPD is related to higher MDA levels that return to normal conditions during the course of treatment. In conclusion, the results suggest that MDA levels can serve as a marker of prognosis and of the success of treatment of the exacerbation of COPD. PMID- 11414509 TI - Sperm parameters changes induced by Azadirachta indica in albino rats. AB - Azadirachta indica treatment for 48 days In albino rats resulted in a decrease in the total sperm count, sperm motility, and forward velocity. The percentage of abnormal sperm increased and the fructose content of caudal semen of the epididymis decreased. The observations suggest that these effects are probably due to an androgen deficiency, caused by the anti-androgenic property of the leaves of A. indica, thereby affecting the physiological maturation of sperm. PMID- 11414510 TI - L-arginine-induced changes in the characteristics of endothelial relaxation. AB - The importance of the functional integrity of endothelium in maintaining vessel tone and the reversibility of its dysfunction by L-arginine (Arg) treatment has made this amino acid very unique and a topic of many experiments. As the behavior of normal vessels against L-Arg was not known properly, we studied the effect of 10(-2) M L-Arg on the dilating effect of Ach and on its characteristics in 10(-6) M phenylephrine-contracted rings prepared from normal rat thoracic aortae, where nitric oxide (NO) mediates most acetylcholine (Ach)-induced relaxation. L arginine attenuated PE-induced contraction, and the pD2 value for phenylephrine shifted to the right from -log M 7.10 +/- 0.1 to -6.67 +/- 0.16. L-arginine supplementation also caused a tremendous change in the characteristics of Ach induced dilation. The share of the indomethacin-sensitive part in the 10(-5) M Ach-induced dilation of L-Arg pretreated rings increased from 26.0 +/- 0.1 to 42.0 +/- 4.6%, whereas the L-NAME-sensitive component decreased from 74.0 +/- 1.6 to 29.0 +/- 3.8% (p<0.001). L-arginine led to the occurrence of a non-NO, non PGI2 component (29.0 +/- 1.1%) in Ach dilation. When regarding the NO-based relaxing response of thoracic aortic rings to Ach in physiological situations, L Arg-induced changes in the components of endothelial derived relaxation seems an important issue to be considered during L-Arg supplementation in the clinic. PMID- 11414511 TI - Exposures of workers at a magazine printing company. PMID- 11414512 TI - Computer resources for planning and responding to chemical emergencies. PMID- 11414513 TI - Injury associated with working near or operating wood chippers. National Institute for Occupational Safety and Health. PMID- 11414514 TI - North American Industry Classification System--a system for the future. PMID- 11414515 TI - Personal exposure of workers to atmospheric PAHs on gasworks sites--The SOLEX study. AB - The aim of the SOLEX study was to estimate the personal exposure of workers to atmospheric polycyclic aromatic hydrocarbons (PAHs) on former gasworks sites in the Paris metropolitan area. Devices to sample gas and particulate phases for 9 PAHs were carried during one working day of a study week in November 1997 by 24 workers and in June 1998 by 19 workers with contrasted job profiles involving different opportunities for contact with the soil; among these volunteers, some were active in the process of contaminated soil remediation during the November study period. PAH concentrations were measured by high-performance liquid chromatography. Subjects working on the site under remediation were clearly more exposed (684.1 ng/m3 for total PAHs in November 1997) than workers, without close contact with soil, whose PAH exposure was similar to that measured by fixed monitors in the ambient air in Paris. The contrasts were weaker in June 1998, when soil remediation had nearly been completed. Only subjects involved in activities with close contact with the ground were found with exposures exceeding background levels. Further investigations are needed to improve our understanding of the influence of the pollutants present in the soil. PMID- 11414516 TI - Exposure to protein aeroallergens in egg processing facilities. AB - Proteinaceous materials in the air can be highly allergenic and result in a range of immunologically mediated respiratory effects, including asthma. We report on the largest evaluation of exposure to date of airborne egg protein concentrations in an egg breaking and processing plant that had cases of occupational asthma. Personal air samples for egg protein were analyzed in duplicate on each PTFE filter using two analytical methods: (1) a commercial assay for non-specific total protein, and (2) indirect competitive inhibition assay using an ELISA method to quantify specific egg protein components. The highest concentrations were found in the egg washing room (mean exposure 644 microg/m3) and breaking room (255 microg/m3), which were also the areas where the risk of being sensitized was the greatest. There was excellent quantitative agreement between the airborne concentrations of total protein and sum of the specific protein antigens (ovalbumin, ovomucoid, and lysozyme). The correlation coefficient of the log-transformed data from the two methods was 0.88 (p < 0.0001). Size-selective sampling also indicated that most of the aerosol was capable of reaching the small airways. The methods described can be utilized to evaluate employee exposure to egg proteins. Exposure documentation, coupled with recommended exposure reduction strategies, could facilitate prevention of future employee sensitization and allergic respiratory responses by identifying high-exposure jobs and evaluating control measures. PMID- 11414517 TI - Task-specific lead exposure during residential lead hazard reduction projects. AB - This article describes a study of worker airborne lead exposure during residential lead hazard reduction projects. The study was performed to characterize short-term, task-specific exposures and to test an airborne lead exposure prediction equation that could assist residential lead abatement contractors in selecting engineering controls and personal protection prior to bidding on a project. A total of 224 personal air samples were collected from 16 workers employed by three contractors at 11 San Francisco Bay-area homes. Exposures ranged from <1.0 microg/m3 to 146.0 microg/m3. Although exposures were low overall (geometric mean = 4.2 microg/m3), there was considerable variability (geometric standard deviation = 4.3 microg/m3). The task-specific approach to exposure assessment used in this study was effective in revealing short-term, high exposures that would be missed during full shift sampling. The highest exposures occurred during paint scraping, demolition, and chemical stripping. Task-specific sampling was also effective in identifying tasks with low exposure potential, such as wet sanding, component removal, and clean-up. Since exposures were not significantly correlated with paint lead content or task area size, two of the variables of the prediction equation, the equation does not appear to be an effective means of predicting airborne lead exposures. Additionally, other factors such as water usage and contractor type had a significant effect on exposure levels. By controlling for these factors, exposures were significantly correlated with the third variable of the prediction equation, the proportion of paint removed from task areas. This study also demonstrates the importance of contractor specific exposure assessments, as there were significant differences in exposures between contractors, even with similar work methods and water use frequency. PMID- 11414518 TI - Survey of the Asp f 1 allergen in office environments. AB - Sick Building Syndrome remains a prevalent problem with patient complaints similar to typical allergy symptoms. Unlike household allergens typically found in domestic reservoirs, the allergen from a common fungus like Aspergillus fumigatus (i.e., Asp f 1) is conceivably widespread in the work environment. This project surveyed airborne levels of the Asp f 1 allergen in office and non industrial occupational environments, as well as the dust reservoirs of A. fumigatus believed to be responsible for those levels. Airborne and bulk dust samples were collected, extracted, and assayed for Asp f 1. Concurrently, bulk dusts collected from the same locations were selectively cultured for A. fumigatus, and mesophilic fungi and bacteria. Samples were collected during both wet and dry climatological conditions from paired wet and dry building locations to examine the possibility of Asp f 1 increases due to fungal growth blooms. Very low levels of Asp f 1 were detected but only in the airborne samples (2/120 positive samples, with 3.6 ng/m3 and 1.8 ng/m3; LOD < 1.2 ng/m3). No dust samples showed even detectable traces of the allergen (LOD = 5 ng/g dust). Although A. fumigatus counts from dusts fluctuated significantly with exterior moisture events, analysis of wet versus dry period samples showed no differences in Asp f 1 levels. These results indicate that even in the presence of measurable fungal concentrations, background levels of Asp f 1 are low. Nonindustrial office buildings devoid of indoor air quality issues were not observed to have significant levels of the Asp f 1 allergen in the geographical region studied. PMID- 11414519 TI - Acute respiratory effects and endotoxin exposure during wheat harvest in Northeastern Colorado. AB - Acute cross-shift respiratory changes were evaluated for workers at 25 farms in northeastern Colorado during the summer of 1994 wheat harvest. Information on workers' respiratory health, past occupational exposures, and smoking status was obtained. Each worker was asked to rank eight acute symptoms before he or she began harvest work for the day. Spirometry was also performed before work began. Each participant wore a high-flow personal air sampling pump for the full shift. At the end of the workshift, spirometry and ranking of the eight acute symptoms were conducted again. Total dust exposure was determined gravimetrically. Total endotoxin was measured by the Limulus amoebocyte lysate (LAL) assay. The 98 harvest workers included in the study ranged in age from 18 to 80. Ten percent of the workers had moderate airway obstruction, as indicated by the pre-shift spirometry test results. Fifty percent of the workers were current or ex-smokers. Despite an unusually poor harvest, total dust exposures ranged from 0.09 to 15.33 mg/m3 (geometric mean 0.83 mg/m3), with 8 percent of workers exposed above the American Conference of Government Industrial Hygienists (ACGIH) threshold limit value (TLV) of 4 mg/m3. Total endotoxin exposures ranged from 4.4 to 744.4 EU/m3 (geometric mean 54.2 EU/m3), with 33 percent of workers exposed above 90 EU/m3, the level suggested as a threshold for acute mucous membrane irritation and pulmonary change among cotton workers. Sixty percent of workers experienced a cross-shift change in at least one respiratory symptom. The respiratory index (sum of cross-shift changes in the eight acute respiratory symptoms) was significantly correlated with both total dust and endotoxin exposure. Cross-shift changes in the spirometric variables were associated with smoking status, age, presence of airway obstruction, and history of chronic respiratory symptoms, but not with dust or endotoxin exposure. Peak expiratory flow rate was found to decrease over the workshift in a manner similar to that experienced by cotton workers. PMID- 11414520 TI - Development of sampling and analytical methods for concerted determination of commonly used chloroacetanilide, chlorotriazine, and 2,4-D herbicides in hand wash, dermal-patch, and air samples. AB - Sampling and analytical methods were developed for commonly used chloroacetanilide, chlorotriazine, and 2,4-D herbicides in hand washes, on dermal patches, and in air. Eight herbicides selected for study were alachlor, atrazine, cyanazine, 2,4-dichlorophenoxyacetic acid (2,4-D), metolachlor, simazine, and two esters of 2,4-D, the 2-butoxyethyl ester (2,4-D, BE) and the 2-ethylhexyl ester (2,4-D, EH). The hand-wash method consisted of shaking the worker's hand in 150 mL of isopropanol in a polyethylene bag for 30 seconds. The dermal-patch method entailed attaching a 10-cm x 10-cm x 0.6-cm polyurethane foam (PUF) patch to the worker for exposure; recovery of the herbicides was achieved by extraction with 40 mL of isopropanol. The air method involved sampling with an OVS-2 tube (which contained an 11-mm quartz fiber filter and two beds of XAD-2 resin) and recovery with 2 mL of 10:90 methanol:methyl t-butyl ether. Analysis of each of the three sample types was performed by gas chromatography with an electron-capture detector. Diazomethane in solution was employed to convert 2,4-D as the free acid to the methyl ester in each of the three methods for ease of gas chromatography. Silicic acid was added to sample solutions to quench excess diazomethane. Limits of detection for all eight herbicides were matrix-dependent and, generally, less than 1 microgram per sample for each matrix. Sampling and analytical methods met NIOSH evaluation criteria for all herbicides in hand-wash samples, for seven herbicides in air samples (all herbicides except cyanazine), and for six herbicides in dermal-patch samples (all herbicides except cyanazine and 2,4-D). Speciation of 2,4-D esters and simultaneous determination of 2,4-D acid were possible without losses of the esters or of other herbicides (acetanilides and triazines) being determined. PMID- 11414521 TI - Limits of viability: dilemmas, decisions, and decision makers. AB - Decision-making about treatments for neonates at the threshold of viability is a complex process that must involve physicians, other health-care professionals, and families. Parents and families bring personal, ideological, cultural, and religious beliefs into their relationship with health-care professionals that have the potential to conflict with professional perceptions of good medical care and the interests of the patient. Neonatologists often find themselves criticized for overtreatment of these extremely premature infants. Yet, from the perspective of the health-care provider, perceived obligations in the face of an uncertain outcome, parental wishes as well as perceptions about legal mandates are often cited as the reasons for the provision of such extraordinary care. Recent reductions in perinatal mortality for premature infants born at the cusp of viability, in conjunction with emerging data on the substantial short- and long term morbidities experienced by infants born between 23-25 weeks' gestation, have engendered a serious debate about professional and parental obligations in the face of extreme uncertainty. The fundamental questions are who ought to be permitted, under the present circumstances of rapidly evolving technologies and innovative therapies, to decide the best interests of the child, and how to achieve consensus regarding treatment goals when the outcome is uncertain and there are divergent views with regard to the infant's best interests. As survival for these infants increases and morbidity remains a significant likelihood, physicians must be cognizant of the power of their technology to impose undesired burdens on these infants. A reasonable, and reasoned, approach for these vulnerable infants requires collaborative decision making incorporating professional recommendations, with an openness, trust and willingness to work with parents to ascertain the best interests of an individual infant. Understanding of and respect for the differing views of the moral obligations of perinatal specialists and families can aid neonatal professionals in resolving interdisciplinary and physician-family conflicts as well as facilitating resolution of neonatal ethical dilemmas. PMID- 11414522 TI - Lack of relationship between variable decelerations during reactive nonstress tests and oligohydramnios. AB - We sought to prospectively determine the value of variable and lambda decelerations noted during nonstress tests in predicting oligohydramnios and perinatal outcome. On the same day as having had reactive nonstress tests, 651 women underwent obstetrical ultrasound examinations for amniotic fluid index (AFI). Lambda and variable fetal heart rate decelerations were identified on sets of otherwise-reactive nonstress tests. Those with variable decelerations composed group 1; those without variable decelerations composed group 2. Demographic, perinatal outcome, and AFI data were compared, and group 1 inclusion criteria were varied. There were 200 nonstress tests in group 1 and 490 in group 2. There was a small difference between groups in maternal age (1.2 years, p = 0.02) and gestational age (0.8 weeks, p = 0.01), but no differences in gravidity, parity, race, or indications for testing (p >0.05). Oligohydramnios was present in 14.5% in group 1 and in 13.3% of group 2 (p = 0.68). In the cases of amniotic fluid index <8 cm, depth and duration of the variable decelerations did not correlate with AFI (R2 = 0.11, p = 0.09). Variable decelerations were associated with intrapartum "fetal distress," but lambda decelerations did not correlate with any outcome. Variable decelerations on an otherwise-reactive nonstress test did not correlate with amniotic fluid volume and were not cause for intervention in this study. PMID- 11414523 TI - Correlation of external and internal monitoring of uterine activity in a cohort of term patients. AB - External tocodynamometry (ET) and intrauterine pressure monitoring (IUPC) are commonly used to assess uterine contractile activity. It is believed that ET does not accurately document contraction amplitude. However, no direct comparison of ET and IUPC has been published in term patients. Our objective was to determine the correlation between frequency, duration, and amplitude as measured by ET and IUPC. Twenty patients between 36 and 41 weeks' gestation were enrolled during active spontaneous labor. Uterine contractions were recorded simultaneously by ET and IUPC for 2 hours. ET and IUPC data were compared using the Spearman correlation coefficient and the Wilcoxon signed-rank test. The median body mass index (BMI) of the patient population was 31.8 kg/m2 (range 22.2 to 42.3). There was good correlation of contraction frequency measured by ET and IUPC (r = 0.75, p = 0.001), but poor correlation of both contraction amplitude (r = 0.26) and duration (r = 0.27). ET and IUPC assessment of uterine contractile activity, specifically contraction amplitude and duration, do not appear to correlate in a population of moderately obese patients. This suggests that ET should not exclusively be relied upon in this population to assess uterine contractile activity during labor. PMID- 11414524 TI - Antifungal susceptibility testing and the correlation with clinical outcome in neonatal candidemia. AB - The objective of this article is to assess the distribution of minimal inhibition concentrations (MIC) for candidal isolates from bloodstreams in neonates and to assess the correlation of clinical outcome with antifungal susceptibility testing. Of the 62 episodes of neonatal candidemia in a Children's Hospital between January 1994 and July 1998, 38 stocked isolates from 38 infants' bloodstreams were available and underwent antifungal susceptibility test according to National Committee for Clinical Laboratory Standards M27-A document. Correlation of clinical response with in vitro results was assessed in 37 patient episode-isolate events. No less than 90% of these isolates tested were susceptible to amphotericin B, flucytosin, and fluconazole. The ranges of amphotericin B MICs and flucytosin MICs were narrow, ranging from 0.25 to 2 microg/mL, respectively. The range of fluconazole MICs was broad, ranging from 0.25 to >64 microg/mL. Successful therapy was achieved in 18 (62%) of 29 amphotericin B-treated patient-episode-susceptible isolate (MIC < or =1 microg/mL) events and 9 (64%) of 14 fluconazole-treated patient-episode susceptible isolate events, respectively. Most isolates from the bloodstreams of neonates with candidemia were susceptible to antifungal agents tested but a low MIC of the antifungal agent did not predict successful therapy in this study. Correlating MICs with clinical outcome in neonatal candidemia requires complex evaluation of other factors. PMID- 11414525 TI - Spontaneous rupture of the liver presenting as scrotal bruising. AB - We describe a case of spontaneous rupture of the liver in a neonate, presenting with features consistent with testicular torsion. Ruptured liver has been well described in neonates following birth trauma, coagulation defects, and Group B Streptococcal infection. The etiology in our case remains undefined. PMID- 11414526 TI - Correlation between prenatal and neonatal birth order in twin pregnancy. AB - Little data is available correlating the in utero order of presentation and the birth order of twins. Our objective was to determine whether birth order in twin pregnancies corresponds to the order of presentation early in pregnancy. All twin pregnancies in which amniocentesis was performed from 1996 to 1998 were identified. Those with discordant genders that delivered at our hospital were included. Order of presentation was documented by ultrasound at the time of amniocentesis. Delivery data were obtained from review of medical records. Statistical comparison was done using two-tailed Fisher's exact test, Student's t test, and Mann-Whitney U test. Sixty patients met inclusion criteria. Birth order corresponded to the order at the time of amniocentesis in 55 of 60 cases (91.7%). There was no difference in the rate of concordance of prenatal and neonatal birth order in twins delivered vaginally compared with those delivered abdominally (90.9 vs. 91.8%, p = 1.0). Cases with discordant prenatal and neonatal birth order had similar maternal ages, gestational ages at amniocentesis and delivery, and fetal presentation at delivery as cases with concordant birth orders. In dichorionic twin pregnancies, birth order is established early in gestation in >90% of cases regardless of route of delivery. PMID- 11414527 TI - Mucinous appendicular cystadenocarcinoma and pregnancy. AB - Primary appendicular adenocarcinoma is rare. We report an unusual mucinous appendicular adenocarcinoma during pregnancy. To our knowledge, this is the second reported case in the literature. A mucinous appendicular cystoadenocarcinoma was discovered unexpectedly and coincidentally during a cesarean section performed in term pregnancy. An appendectomy was simultaneously performed, followed by a right hemicolectomy. Given the exceptionally low incidence tumors co-existing with pregnancy, we recommend a careful and complete appendix examination during cesarean and postpartum sterilization, and subsequent removal of tumors should they exist. PMID- 11414528 TI - DNA diagnosis and management of Hermansky-Pudlak syndrome in pregnancy. AB - Hermansky-Pudlak syndrome (HPS) is an autosomal recessive disorder characterized by oculocutaneous albinism (OCA), platelet storage pool deficiency, and ceroid lipofuscin deposition. Sequelae including pulmonary fibrosis, colitis, and hemorrhagic diathesis can impact obstetric management. An 18-year-old primigravida with OCA was diagnosed during pregnancy with Hermansky-Pudlak syndrome by DNA analysis. Uneventful vaginal delivery occurred at term following prophylactic platelet transfusion. Women of northwestern Puerto Rican descent with OCA should be offered testing for HPS. Identification of affected individuals may permit optimal obstetric management. PMID- 11414529 TI - Cardiomyopathy in pregnancy: a retrospective study. AB - Women with dilated cardiomyopathies (DCM) have traditionally been advised not to attempt to carry pregnancies. This is largely based on data derived from studies of the course of peripartum cardiomyopathy (PPCM) and it is not clear that this extrapolation is appropriate. Our objective was to compare maternal and obstetric outcomes of pregnancies in women with preexisting DCM to women with PPCM. A 10 year retrospective comparative cohort study was undertaken of women between the ages of 15 and 40 discharged from two university medical centers with the diagnosis of cardiomyopathy (CM). Patients were included in the study if CM was diagnosed prior to pregnancy (DCM group) or if CM developed during pregnancy or within 5 months postpartum (PPCM group), and follow-up data was available. Thirty one subjects were included in the study. Twenty-three women with the diagnosis of PPCM were compared with 8 women with DCM of other etiologies. There were no significant differences in maternal age, race, parity, tobacco or other substance use, or other risk factors between the two groups. Maternal outcomes in the PPCM group were significantly worse than in the DCM group, with three maternal deaths and four women undergoing heart transplants (p = 0.05). In the DCM group, one woman with a prepregnancy ejection fraction of 16% underwent transplantation after termination of pregnancy for genetic indications. None of the other women in the DCM group had a significant decline in cardiac status. Infant outcomes in both groups were uniformly good. PPCM represents an acute, evolving insult to the pregnant or postpartum woman. The prognosis of this condition should not be used for counseling women with DCM considering pregnancy. Women with stable DCM may do well during pregnancy without significant deterioration in their cardiac status. PMID- 11414530 TI - A prospective controlled trial of albuterol aerosol delivered via metered dose inhaler-spacer device (MDI) versus jet nebulizer in ventilated preterm neonates. AB - The objective of this study was to identify the most efficient and cost-effective nebulizer device for delivery of albuterol aerosol as a bronchodilator in ventilated preterm infants. Bronchodilators are frequently used as part of the therapeutic regimen of ventilated preterm infants. This can be delivered by different types of nebulizers like the Jet or metered dose inhaler (MDI) spacer device. Fifty-three premature infants being ventilated for RDS (24 to 34 weeks of gestation) were studied just prior to extubation. Twenty-four of them received standard doses of albuterol aerosol via Jet nebulizer and 29 via MDI-spacer. Heart rate, respiratory rate, oxygen saturation, lung compliance, and airway resistance were monitored prior and 15 minutes after albuterol delivery. There were significant changes in the parameters studied between pre- and postnebulizer treatment. In both groups, there was a significant improvement in lung function as evidenced by 13-24% decreased airway resistance (RAWE) and 3-7% increased lung compliance (CDYN). There was also a beneficial clinical response as demonstrated by increased oxygen saturations. These findings suggest that both MDI-spacer and Jet nebulizer are equally effective in delivering the albuterol aerosol to the lower respiratory tract. Since a small dose of albuterol delivered via the MDI spacer improved lung function as effectively as a higher dose via the Jet nebulizer, the MDI-spacer would be the preferred mode of aerosol administration, especially because it takes only 2 minutes to deliver it. Furthermore, it was also cost-effective as one MDI-spacer treatment costs 2 cents, while a Jet treatment costs 10 cents in our neonatal intensive care unit (NICU). PMID- 11414531 TI - Multi-stage model estimates of lung cancer risk from exposure to diesel exhaust, based on a U.S. railroad worker cohort. Response to commentary. PMID- 11414532 TI - Risk analysis, systems analysis, and Covey's seven habits. AB - This article relates problem solving to the common approaches of the gestalt holistic philosophies of systems analysis, risk analysis, and Stephen Covey's Seven Habits of Highly Effective People. Guiding principles developed on the basis of these philosophies provide the foundations for methodological frameworks that build on a plethora of theory, methods, tools, and techniques. Although systems analysis and risk analysis differ in their historical evolution and technical maturity, both study and solve problems using methodological frameworks that share a holistic vision. PMID- 11414533 TI - Robustness of conditional moments: an application to premium calculation of reinsurance treaties. AB - In this study, the tail probability of a class of distributions commonly used in assessing the severity of insurance losses was examined. Without specifying any particular distribution, the use of an algebraic functional form Cx(-alpha) to approximate the tail behavior of the distributions in the class was demonstrated. Norwegian fire insurance data were examined, and the algebraic functional form was applied to derive the expected loss of a reinsurance treaty that covers all losses exceeding a retention limit. It was shown that (1) the expected loss is insensitive to the parameter alpha for a high retention limit (e.g., a catastrophe treaty), and (2) with a low retention limit (e.g., a largest claim treaty), a reliable estimate of the parameter alpha and a sound judgment on the maximum potential loss of the treaty could provide useful and defensible summary statistics for pricing the treaty. Thus, when dealing with the losses of certain reinsurance treaties, it was concluded that knowledge of a specific probability distribution is not critical, and the summary statistics derived from the model are robust with respect to a large class of loss distributions. PMID- 11414534 TI - Property loss estimation for wind and earthquake perils. AB - This article describes the development of a generic loss assessment methodology, which is applicable to earthquake and windstorm perils worldwide. The latest information regarding hazard estimation is first integrated with the parameters that best describe the intensity of the action of both windstorms and earthquakes on building structures, for events with defined average return periods or recurrence intervals. The subsequent evaluation of building vulnerability (damageability) under the action of both earthquake and windstorm loadings utilizes information on damage and loss from past events, along with an assessment of the key building properties (including age and quality of design and construction), to assess information about the ability of buildings to withstand such loadings and hence to assign a building type to the particular risk or portfolio of risks. This predicted damage information is then translated into risk-specific mathematical vulnerability functions, which enable numerical evaluation of the probability of building damage arising at various defined levels. By assigning cost factors to the defined damage levels, the associated computation of total loss at a given level of hazard may be achieved. This developed methodology is universal in the sense that it may be applied successfully to buildings situated in a variety of earthquake and windstorm environments, ranging from very low to extreme levels of hazard. As a loss prediction tool, it enables accurate estimation of losses from potential scenario events linked to defined return periods and, hence, can greatly assist risk assessment and planning. PMID- 11414535 TI - Ecological risk assessment framework for low-altitude aircraft overflights: I. Planning the analysis and estimating exposure. AB - An ecological risk assessment framework for low-altitude aircraft overflights was developed, with special emphasis on military applications. The problem formulation and exposure analysis phases are presented in this article; an analysis of effects and risk characterization is presented in a companion article. The intent of this article is threefold: (1) to illustrate the development of a generic framework for the ecological risk assessment of an activity, (2) to show how the U.S. Environmental Protection Agency's ecological risk assessment paradigm can be applied to an activity other than the release of a chemical, and (3) to provide guidance for the assessment of ecological risks from low-altitude aircraft overflights. The key stressor for low-altitude aircraft overflights is usually sound, although visual and physical (collision) stressors may also be significant. Susceptible and regulated wildlife populations are the major assessment endpoint entities, although plant communities may be impacted by takeoffs and landings. The exposure analysis utilizes measurements of wildlife locations, measurements of sound levels at the wildlife locations, measurements of slant distances from aircraft to wildlife, models that extrapolate sound from the source aircraft to the ground, and bird-strike probability models. Some of the challenges to conducting a risk assessment for aircraft overflights include prioritizing potential stressors and endpoints, choosing exposure metrics that relate to wildlife responses, obtaining good estimates of sound or distance, and estimating wildlife locations. PMID- 11414536 TI - Ecological risk assessment framework for low-altitude aircraft overflights: II. Estimating effects on wildlife. AB - An ecological risk assessment framework for aircraft overflights has been developed, with special emphasis on military applications. This article presents the analysis of effects and risk characterization phases; the problem formulation and exposure analysis phases are presented in a companion article. The framework addresses the effects of sound, visual stressors, and collision on the abundance and production of wildlife populations. Profiles of effects, including thresholds, are highlighted for two groups of endpoint species: ungulates (hoofed mammals) and pinnipeds (seals, sea lions, walruses). Several factors complicate the analysis of effects for aircraft overflights. Studies of the effects of aircraft overflights previously have not been associated with a quantitative assessment framework; therefore no consistent relations between exposure and population-level response have been developed. Information on behavioral effects of overflights by military aircraft (or component stressors) on most wildlife species is sparse. Moreover, models that relate behavioral changes to abundance or reproduction, and those that relate behavioral or hearing effects thresholds from one population to another are generally not available. The aggregation of sound frequencies, durations, and the view of the aircraft into the single exposure metric of slant distance is not always the best predictor of effects, but effects associated with more specific exposure metrics (e.g., narrow sound spectra) may not be easily determined or added. The weight of evidence and uncertainty analyses of the risk characterization for overflights are also discussed in this article. PMID- 11414537 TI - Multimedia analysis of PAHs and nitro-PAH daughter products in the Los Angeles Basin. AB - Polycyclic aromatic hydrocarbons (PAHs) that are released into the atmosphere may have health consequences that can be compounded by their nitro-PAH atmospheric transformation products. The available literature suggests that some of the atmospheric nitro-PAH daughter products may increase the overall environmental health risk associated with PAHs. Therefore, an important issue is whether there is merit in considering atmospheric transformation products of air toxins when conducting environmental health-risk analyses. To illustrate the above issue, a comparative analysis of the potential risk that may be imposed by PAHs and their daughter products was carried out for the Los Angeles Basin. The analysis consisted of first assessing the multimedia environmental concentration of selected PAHs and nitro-PAHs using a spatial-compartmental modeling approach coupled with available monitoring data. Multimedia concentrations were then used to estimate chemical media-specific mutagenic densities as well as average daily intake from multiple pathways, followed by cancer risk for the known carcinogens among the study chemicals. The analysis revealed that mutagenic densities of the nitro-PAH daughter products can significantly exceed those of the parent PAHs. The results of this study suggest that there is merit in further investigation of the potential contribution of nitro-PAHs to the overall environmental health risk associated with airborne PAHs. PMID- 11414538 TI - A weighted composite dose-response model for human salmonellosis. AB - This article describes the development of a weighted composite dose-response model for human salmonellosis. Data from previously reported human challenge studies were categorized into two different groups representing low and moderately virulent/pathogenic Salmonella strains based on a disease end point. Because epidemiological data indicate that some Salmonella strains are particularly pathogenic, and in the absence of human feeding study data for such strains, Shigella dysenteriae was used as a proxy for highly virulent strains. Three single-hit dose-response models were applied to the human feeding study data and evaluated for best fit using maximum likelihood estimation: (1) the exponential (E-1pop), (2) the two-subpopulation exponential (E-2pop), and (3) the Beta-Poisson (BP). Based on the goodness-of-fit test, the E-1pop and BP were the best-fit models for low and moderately virulent/pathogenic Salmonella strains, and the E-2pop and BP models were better for highly virulent/pathogenic strains. Epistemic analysis was conducted by determining the degree of confidence associated with the selected models, which was found to be 50%/50% (E-1pop/BP) for low and moderately pathogenic Salmonella strains, and 9.8%/90.2% (E-2pop/BP) for highly virulent strains. The degree of confidence for each component model and variations in the proportion of strains within each virulence/pathogenicity category were incorporated into the overall composite model. This study describes the influence of variation in strain virulence and host susceptibility on the shape of the population dose-response relationship. PMID- 11414539 TI - Socioeconomic determinants of health- and food safety-related risk perceptions. AB - Individual and societal perceptions of food-related health risks are multidimensional and complex. Social, political, psychological, and economic factors interact with technological factors and affect perceptions in complex ways. Previous research found that the significant determinants of risk perceptions include socioeconomic and behavioral variables. Most of these past results are based on two-way comparisons and factor analysis. The objective of this study was to analyze the significance of socioeconomic determinants of risk perceptions concerning health and food safety. A multivariate approach was used and the results were compared with earlier bivariate results to determine which socioeconomic predictors were robust across methods. There were two major findings in this study. The first was that the results in the multivariate models were generally consistent with earlier bivariate analysis. That is, variables such as household income, number of children, gender, age, and voting preferences were strong predictors of an individual's risk perceptions. The second result was that the gender of the respondent was the only variable found to be robust across all three classes of health and food safety issues across two time periods. PMID- 11414540 TI - Perceived risks of conventional and organic produce: pesticides, pathogens, and natural toxins. AB - Public risk perceptions and demand for safer food are important factors shaping agricultural production practices in the United States. Despite documented food safety concerns, little attempt has been made to elicit consumers' subjective risk judgments for a range of food safety hazards or to identify factors most predictive of perceived food safety risks. In this study, over 700 conventional and organic fresh produce buyers in the Boston area were surveyed for their perceived food safety risks. Survey results showed that consumers perceived relatively high risks associated with the consumption and production of conventionally grown produce compared with other public health hazards. For example, conventional and organic food buyers estimated the median annual fatality rate due to pesticide residues on conventionally grown food to be about 50 per million and 200 per million, respectively, which is similar in magnitude to the annual mortality risk from motor vehicle accidents in the United States. Over 90% of survey respondents also perceived a reduction in pesticide residue risk associated with substituting organically grown produce for conventionally grown produce, and nearly 50% perceived a risk reduction due to natural toxins and microbial pathogens. Multiple regression analyses indicate that only a few factors are consistently predictive of higher risk perceptions, including feelings of distrust toward regulatory agencies and the safety of the food supply. A variety of factors were found to be significant predictors of specific categories of food hazards, suggesting that consumers may view food safety risks as dissimilar from one another. Based on study findings, it is recommended that future agricultural policies and risk communication efforts utilize a comparative risk approach that targets a range of food safety hazards. PMID- 11414541 TI - People's preferences for safety control: why does baseline risk matter? AB - Previous tests of how people's valuations of safety vary with the level of baseline risk have left policymakers with a rather mixed message. Some data support the conventional hypothesis derived from economic theory that marginal valuations of risk changes increase with the baseline level, but other data reject it. With these indeterminate findings in mind, the present study adopted an in-depth quantitative-qualitative methodology to explore the preferences of a general population sample (N = 147) for safety programs targeted at hazards with different age groups at risk and numbers of deaths per year. The data clearly showed that self- or household interest mattered to people (e.g., programs in which the respondent or their household members were in the at-risk age group were evaluated more positively). The number of deaths also mattered, although not always for reasons strictly consistent with the conventional hypothesis. Rather, the qualitative findings suggested that evaluations may be driven by auxiliary assumptions about aspects of the programs induced by this information (e.g., people believe that a higher number of deaths is an indicator that more people are likely to be exposed to that hazard). These findings bring into question the extent to which quantitative responses from stated preference surveys can be taken at face value to form a reliable basis for public policy. PMID- 11414542 TI - Differences in expert and lay judgments of risk: myth or reality? AB - This article evaluates the nine empirical studies that have been conducted on expert versus lay judgments of risk. Contrary to received wisdom, this study finds that there is little empirical evidence for the propositions (1) that experts judge risk differently from members of the public or (2) that experts are more veridical in their risk assessments. Methodological weaknesses in the early research are documented, and it is shown that the results of more recent studies are confounded by social and demographic factors that have been found to correlate with judgments of risk. Using a task-analysis taxonomy, a template is provided for the documentation of future studies of expert-lay differences/similarities that will facilitate analytic comparison. PMID- 11414543 TI - Evaluating the effectiveness of risk-reduction strategies for consumer chemical products. AB - Communication about risks offers a voluntary approach to reducing exposure to pollutants. Its adequacy depends on its impact on behavior. Estimating those impacts first requires characterizing current activities and their associated risk levels, and then predicting the effectiveness of risk-reduction strategies. Characterizing the risks from chemical consumer products requires knowledge of both the physical and the behavioral processes that influence exposures. This article presents an integrated approach that combines consumer interviews, users' beliefs and behaviors, and quantitative exposure modeling. This model was demonstrated in the context of consumer exposure to a methylene chloride-based paint stripper, showing how it could be used to evaluate current levels of risk and predict the effectiveness of proposed voluntary risk-reduction strategies. PMID- 11414544 TI - Mitigation and benefits measures as policy tools for siting potentially hazardous facilities: determinants of effectiveness and appropriateness. AB - How do mitigation and benefits measures affect public acceptance for siting different kinds of potentially hazardous facilities? What kinds of benefits measures are seen as most (or least) appropriate for different kinds of facilities? This study used a nationwide telephone survey consisting of 1,234 interviews with randomly selected respondents to test for the effects of packages of safety and benefits measures for siting a landfill, prison, incinerator and nuclear waste repository. The experimental design used in the survey permits analysis of the fractions of respondents who are willing to change their initial levels of acceptance (or opposition) when presented with a sequence of the safety and benefit measures. The measures vary significantly in their impact on levels of acceptance for the facilities, and some measures that would at face value appear to reassure residents of facility safety turn out to lack credibility and therefore diminish facility acceptance. Ordering of the benefits versus safety measures significantly affects changes in acceptance in surprising ways. The perceived appropriateness of different kinds of benefits measures varies systematically by the type of facility under consideration. It appears that successful benefits packages will directly address the underlying dimensions of concern caused by the facility. These findings point to the importance of further research on "commensurable" benefits measures. PMID- 11414545 TI - Temporal variability and ignorance in Monte Carlo contaminant bioaccumulation models: a case study with selenium in Mytilus edulis. AB - Although the parameters for contaminant bioaccumulation models most likely vary over time, lack of data makes it impossible to quantify this variability. As a consequence, Monte Carlo models of contaminant bioaccumulation often treat all parameters as having fixed true values that are unknown. This can lead to biased distributions of predicted contaminant concentrations. This article demonstrates this phenomenon with a case study of selenium accumulation in the mussel Mytilus edulis in San Francisco Bay. "Ignorance-only" simulations (in which phytoplankton and bioavailable selenium concentrations are constant over time, but sampled from distributions of field measurements taken at different times), which an analyst might be forced to use due to lack of data, were compared with "variability and ignorance" simulations (sampling phytoplankton and bioavailable selenium concentrations each month). It was found that ignorance-only simulations may underestimate or overestimate the median predicted contaminant concentration at any time, relative to variability and ignorance simulations. However, over a long enough time period (such as the complete seasonal cycle in a seasonal model), treating temporal variability as if it were ignorance at least gave a range of predicted concentrations that enclosed the range predicted by explicit treatment of temporal variability. Comparing the temporal variability in field data with that predicted by simulations may indicate whether the right amount of temporal variability is being included in input variables. Sensitivity analysis combined with biological knowledge suggests which parameters might make important contributions to temporal variability. Temporal variability is potentially more complicated to deal with than other types of stochastic variability, because of the range of time scales over which parameters may vary. PMID- 11414546 TI - Generalized anxiety disorder: nature and course. AB - Generalized anxiety disorder (GAD) is a chronic and highly prevalent disorder in the adult population, yet it remains a relatively poorly understood condition. Clinicians may be familiar with the symptoms of enduring excessive worrying, anxiety, and hypervigilance that are characteristic of GAD, but may not necessarily recognize that these are usually symptoms of a distinct psychiatric disorder. Despite changes in diagnostic criteria, estimates of prevalence for GAD are remarkably consistent across epidemiologic studies. Lifetime prevalence in the general population is estimated at 5% (DSM-III and/or DSM-III-R criteria), with rates as high as 10% among women aged 40 years and above, and cross sectional rates among primary care attenders are about 8%, making GAD the most prevalent anxiety disorder in primary care. The age at onset of GAD differs from that of other anxiety disorders: prevalence rates are low in adolescents and young adults but increase substantially with age. Females are at greater risk than males, and the disorder is correlated with being unemployed or a housewife or having a chronic medical illness. GAD is frequently associated with comorbid depression and other anxiety and somatoform disorders. Significant GAD-specific disability occurs even when comorbidity is not present. PMID- 11414547 TI - Neurobiological mechanisms in generalized anxiety disorder. AB - Generalized anxiety disorder (GAD) is a common and serious disorder. Despite this fact, there is no clear understanding of the exact neurobiological changes underlying the condition. To date, there are few studies of neurobiological function in patients with GAD, and only limited comparative data with depression are available. Advances in neuroanatomical imaging techniques are beginning to allow detailed study of regional blood flow and metabolism and may offer insights into the specific regions of the brain involved in GAD. Investigations into neurotransmitter dysfunction have implicated the gamma-aminobutyric acid/benzodiazepine, serotonergic, and noradrenergic systems in this disorder. Variations in sleep patterns have also been assessed and indicate a biological separation from depression. PMID- 11414548 TI - Comorbidity in generalized anxiety disorder: impact and implications. AB - Studying comorbidity may be particularly useful in shedding light on the nature, course, and management of generalized anxiety disorder (GAD). This article reviews the common comorbidities in GAD, their impact on functioning and on medical utilization, and their implications for treatment. Comorbidity in this area is complex in that GAD may be primary (predating other illnesses) or secondary (following other conditions). Nevertheless, studies demonstrate the high prevalence of patients with a matrix of anxious-somatic-depressive symptoms and show that GAD comorbidity is associated with both increased disability and increased medical utilization. Clinical implications of this work include the need for rigorous assessment of anxiety, somatic, and depressive symptoms in patients who present with any one of these sets of complaints and the possibility that early treatment of GAD may be critical in preventing subsequent morbidity. PMID- 11414549 TI - Psychotherapy for generalized anxiety disorder. AB - The present article describes the basic therapeutic techniques used in the cognitive-behavioral therapy (CBT) of generalized anxiety disorders and reviews the methodological characteristics and outcomes of 13 controlled clinical trials. The studies in general display rigorous methodology, and their outcomes are quite consistent. CBT has been shown to yield clinical improvements in both anxiety and depression that are superior to no treatment and nonspecific control conditions (and at times to either cognitive therapy alone or behavioral therapy alone) at both posttherapy and follow-up. CBT is also associated with low dropout rates, maintained long-term improvements, and the largest within-group and between-group effect sizes relative to all other comparison conditions. PMID- 11414550 TI - What is generalized anxiety disorder? AB - Generalized, persistent, and free-floating anxiety was first described by Freud in 1894, although the diagnostic term generalized anxiety disorder (GAD) was not included in classification systems until 1980 (Diagnostic and Statistical Manual for Mental Disorders, Third Edition [DSM-III]). Initially considered a residual category to be used when no other diagnosis could be made, it is now widely accepted that GAD represents a distinct diagnostic category. Since 1980, revisions to the diagnostic criteria for GAD in the DSM-III-R and DSM-IV classifications have markedly redefined this disorder, increasing the duration criterion to 6 months and increasing the emphasis on worry and psychic symptoms. This article reviews the development of the diagnostic criteria for defining GAD from Freud to DSM-IV and compares the DSM-IV criteria with the criteria set forth in the tenth revision of the International Classification of Diseases. The impact of the changes in diagnostic criteria on research into GAD, and on diagnosis, differential diagnosis, and treatment of GAD, will be discussed. PMID- 11414551 TI - Pharmacotherapy of generalized anxiety disorder. AB - Benzodiazepines have traditionally been used to treat acute anxiety disorders, but they are not ideal in the treatment of chronic generalized anxiety disorder (GAD). Following long-term therapy, benzodiazepines have the potential to produce dependency and withdrawal symptoms. In addition, although agents such as the benzodiazepines and buspirone alleviate anxiety, they have little effect on depression, which is a common comorbidity of GAD. Antidepressants have long been viewed as promising alternatives to benzodiazepines for the treatment of some types of anxiety. Although they have been shown to be useful in the treatment of panic disorder, social anxiety disorder/social phobia, and obsessive-compulsive disorder, they have not until recently been regarded as potential therapies for GAD. Treatment with antidepressants has opened up a new area of investigation into the pharmacotherapy of GAD, with a growing body of evidence supporting the role of therapies such as paroxetine and venlafaxine extended release. PMID- 11414552 TI - Consensus statement on generalized anxiety disorder from the International Consensus Group on Depression and Anxiety. AB - OBJECTIVE: To provide primary care clinicians with a better understanding of management issues in generalized anxiety disorder (GAD) and guide clinical practice with recommendations on the appropriate treatment strategy. PARTICIPANTS: The 4 members of the International Consensus Group on Depression and Anxiety were James C. Ballenger (chair), Jonathan R.T. Davidson, Yves Lecrubier, and David J. Nutt. Four additional faculty members invited by the chair were Karl Rickels, Hans-Ulrich Wittchen, Dan J. Stein, and Thomas D. Borkovec. EVIDENCE: The consensus statement is based on the 6 review articles that are published in this supplement and the scientific literature relevant to the issues reviewed in these articles. CONSENSUS PROCESS: Group meetings were held over a 2-day period. On day 1, the group discussed the review articles and the chair identified key issues for further debate. On day 2, the group discussed these issues to arrive at a consensus view. After the group meetings, the consensus statement was drafted by the chair and approved by all attendees. CONCLUSIONS: GAD is the most common anxiety disorder in primary care and is highly debilitating. Furthermore, it is frequently comorbid with depression and other anxiety disorders, which exacerbates functional impairment. Antidepressants (serotonin reuptake inhibitors, serotonin-norepinephrine reuptake inhibitors, and nonsedating tricyclic antidepressants) are generally the most appropriate first line pharmacotherapy for GAD, since they are also effective against comorbid psychiatric disorders and are suitable for long-term use. Cognitive-behavioral therapy is the preferred form of psychotherapy for GAD, although when GAD is comorbid with depression, pharmacotherapy is increasingly indicated. PMID- 11414553 TI - Addressing an idiosyncrasy in estimating survival curves using double sampling in the presence of self-selected right censoring. AB - We investigate the use of follow-up samples of individuals to estimate survival curves from studies that are subject to right censoring from two sources: (i) early termination of the study, namely, administrative censoring, or (ii) censoring due to lost data prior to administrative censoring, so-called dropout. We assume that, for the full cohort of individuals, administrative censoring times are independent of the subjects' inherent characteristics, including survival time. To address the loss to censoring due to dropout, which we allow to be possibly selective, we consider an intensive second phase of the study where a representative sample of the originally lost subjects is subsequently followed and their data recorded. As with double-sampling designs in survey methodology, the objective is to provide data on a representative subset of the dropouts. Despite assumed full response from the follow-up sample, we show that, in general in our setting, administrative censoring times are not independent of survival times within the two subgroups, nondropouts and sampled dropouts. As a result, the stratified Kaplan-Meier estimator is not appropriate for the cohort survival curve. Moreover, using the concept of potential outcomes, as opposed to observed outcomes, and thereby explicitly formulating the problem as a missing data problem, reveals and addresses these complications. We present an estimation method based on the likelihood of an easily observed subset of the data and study its properties analytically for large samples. We evaluate our method in a realistic situation by simulating data that match published margins on survival and dropout from an actual hip-replacement study. Limitations and extensions of our design and analytic method are discussed. PMID- 11414554 TI - Discussion of the Frangakis and Rubin article. PMID- 11414555 TI - Discussion of double sampling for survival analysis. PMID- 11414556 TI - A model for a proportional treatment effect on disease progression. AB - Treatments intended to slow the progression of chronic diseases are often hypothesized to reduce the rate of further injury to a biological system without improving the current level of functioning. In this situation, the treatment effect may be negligible for patients whose disease would have been stable without the treatment but would be expected to be an increasing function of the progression rate in patients with worsening disease. This article considers a variation of the Laird Ware mixed effects model in which the effect of the treatment on the slope of a longitudinal outcome is assumed to be proportional to the progression rate for patients with progressive disease. Inference based on maximum likelihood and a generalized estimating equations procedure is considered. Under the proportional effect assumption, the precision of the estimated treatment effect can be increased by incorporating the functional relationship between the model parameters and the variance of the outcome variable, particularly when the magnitude of the mean slope of the outcome is small compared with the standard deviation of the slopes. An example from a study of chronic renal disease is used to illustrate insights provided by the proportional effect model that may be overlooked with models assuming additive treatment effects. PMID- 11414557 TI - Using weighted Kaplan-Meier statistics in nonparametric comparisons of paired censored survival outcomes. AB - This research introduces methods for nonparametric testing of weighted integrated survival differences in the context of paired censored survival designs. The current work extends work done by Pepe and Fleming (1989, Biometrics 45, 497 507), which considered similar test statistics directed toward independent treatment group comparisons. An asymptotic closed-form distribution of the proposed family of tests is presented, along with variance estimates constructed under null and alternative hypotheses using nonparametric maximum likelihood estimates of the closed-form quantities. The described method allows for additional information from individuals with no corresponding matched pair member to be incorporated into the test statistic in sampling scenarios where singletons are not prone to selection bias. Simulations presented over a range of potential dependence in the paired censored survival data demonstrate substantial power gains associated with taking into account the dependence structure. Consequences of ignoring the paired nature of the data include overly conservative tests in terms of power and size. In fact, simulation results using tests for independent samples in the presence of positive correlation consistently undershot both size and power targets that would have been attained in the absence of correlation. This additional worrisome effect on operating characteristics highlights the need for accounting for dependence in this popular family of tests. PMID- 11414558 TI - Nonparametric tests for the gap time distributions of serial events based on censored data. AB - This article deals with the problem of comparing two populations with respect to the distribution of the gap time between two successive events when each subject can experience a series of events and when the event times are potentially right censored. Several families of nonparametric tests are developed, all of which allow arbitrary distributions and dependence structures for the serial events. The asymptotic and small-sample properties of the proposed tests are investigated. An illustration with data taken from a colon cancer study is provided. The related problem of testing the independence of two successive gap times is also studied. PMID- 11414559 TI - Joint regression analysis of survival and quality-adjusted survival. AB - In this paper, a semiparametric bivariate linear regression model for survival and quality-adjusted survival is investigated. Even with a parametric specification for the joint, distribution, maximum likelihood is not applicable because of induced informative censoring. We propose inference procedures based on estimating functions. The estimators are consistent and asymptotically normal. Hypothesis tests and confidence intervals may be constructed with easy-to implement resampling techniques. Simultaneous regression modeling of survival and quality-adjusted survival has not been studied formally. Our methodology gives parameter estimates that are highly interpretable in the context of a cost effectiveness analysis. The usefulness of the proposal is illustrated with a breast cancer dataset. PMID- 11414560 TI - Bayesian semiparametric models for survival data with a cure fraction. AB - We propose methods for Bayesian inference for a new class of semiparametric survival models with a cure fraction. Specifically, we propose a semiparametric cure rate model with a smoothing parameter that controls the degree of parametricity in the right tail of the survival distribution. We show that such a parameter is crucial for these kinds of models and can have an impact on the posterior estimates. Several novel properties of the proposed model are derived. In addition, we propose a class of improper noninformative priors based on this model and examine the properties of the implied posterior. Also, a class of informative priors based on historical data is proposed and its theoretical properties are investigated. A case study involving a melanoma clinical trial is discussed in detail to demonstrate the proposed methodology. PMID- 11414561 TI - Estimation and prediction for cancer screening models using deconvolution and smoothing. AB - The model that specifies that cancer incidence, I, is the convolution of the preclinical incidence, g, and the density of time in the preclinical phase, f, has frequently been utilized to model data from cancer screening trials and to estimate such quantities as sojourn time, lead time, and sensitivity. When this model is fit to the above data, the parameters of f as well as the parameter(s) governing screening sensitivity must be estimated. Previously, g was either assumed to be equal to clinical incidence or assumed to be a constant or exponential function that also had to be estimated. Here we assume that the underlying incidence, I, in the study population (in the absence of screening) is known. With I known, g then becomes a function of f, which can be solved for using (numerical) deconvolution, thus eliminating the need to estimate g or make assumptions about it. Since numerical deconvolution procedures may be highly unstable, however, we incorporate a smoothing procedure that produces a realistic g function while still closely reproducing the original incidence function I upon convolution with f. We have also added the concept of competing mortality to the convolution model. This, along with the realistic preclinical incidence function described above, results in more accurate estimates of sojourn time and lead time and allows for estimation of quantities related to overdiagnosis, which we define here. PMID- 11414562 TI - A flexible parametric model for combining current status and age at first diagnosis data. AB - In some cross-sectional studies of chronic disease, data consist of the age at examination, whether the disease was present at the exam, and recall of the age at first diagnosis. This article describes a flexible parametric approach for combining current status and age at first diagnosis data. We assume that the log odds of onset by a given age and of detection by a given age conditional on onset by that age are nondecreasing functions of time plus linear combinations of covariates. Piecewise linear models are used to characterize changes across time in the baseline odds. Methods are described for accommodating informatively missing current status data and inferences based on the age-specific incidence of disease prior to a landmark event (e.g., puberty, menopause). Our formulation enables straightforward maximum likelihood estimation without requiring restrictive parametric or Markov assumptions. The methods are applied to data from a study of uterine fibroids. PMID- 11414563 TI - Inference in randomized studies with informative censoring and discrete time-to event endpoints. AB - In this article, we present a method for estimating and comparing the treatment specific distributions of a discrete time-to-event variable from right-censored data. Our method allows for (1) adjustment for informative censoring due to measured prognostic factors for time to event and censoring and (2) quantification of the sensitivity of the inference to residual dependence between time to event and censoring due to unmeasured factors. We develop our approach in the context of a randomized trial for the treatment of chronic schizophrenia. We perform a simulation study to assess the practical performance of our methodology. PMID- 11414564 TI - Augmented inverse probability weighted estimator for Cox missing covariate regression. AB - This article investigates an augmented inverse selection probability weighted estimator for Cox regression parameter estimation when covariate variables are incomplete. This estimator extends the Horvitz and Thompson (1952, Journal of the American Statistical Association 47, 663-685) weighted estimator. This estimator is doubly robust because it is consistent as long as either the selection probability model or the joint distribution of covariates is correctly specified. The augmentation term of the estimating equation depends on the baseline cumulative hazard and on a conditional distribution that can be implemented by using an EM-type algorithm. This method is compared with some previously proposed estimators via simulation studies. The method is applied to a real example. PMID- 11414565 TI - Nonoverlapping clusters: approximate distribution and application to molecular biology. AB - An approach is developed for the screening of genomic sequence data to identify gene regulatory regions. This approach is based on deciding if putative transcription factor binding sites are clustered together to a greater extent than one would expect by chance. Given n events occurring on an interval of width L (L base pairs), an r:w cluster is defined as r + 1 consecutive events all contained within a window of length wL. Accurate and easily computable approximations are derived for the distribution of the number of nonoverlapping r:w clusters under the model that the positions of the n events have a uniform distribution. Simulations demonstrate that these approximations have greater accuracy than existing methods. The approximation is applied to detect erythroid specific regulatory regions in genomic DNA sequences, first in an artificial case where r is specified a priori and then as part of an exploratory approach. PMID- 11414566 TI - Estimating the frequency distribution of crossovers during meiosis from recombination data. AB - Estimation of tetrad crossover frequency distributions from genetic recombination data is a classic problem dating back to Weinstein (1936, Genetics 21, 155-199). But a number of important issues, such as how to specify the maximum number of crossovers, how to construct confidence intervals for crossover probabilities, and how to obtain correct p-values for hypothesis tests, have never been adequately addressed. In this article, we obtain some properties of the maximum likelihood estimate (MLE) for crossover probabilities that imply guidelines for choosing the maximum number of crossovers. We give these results for both normal meiosis and meiosis with nondisjunction. We also develop an accelerated EM algorithm to find the MLE more efficiently. We propose bootstrap-based methods to find confidence intervals and p-values and conduct simulation studies to check the validity of the bootstrap approach. PMID- 11414567 TI - Identifying chromosomal fragile sites from a hierarchical-clustering point of view. AB - Identification of fragile sites is a way to investigate the genetic abnormalities that are the hallmark of cancer and play an important role in carcinogenesis. Manifestation of nonrandom breakage at a chromosome band is an essential criterion for determination of the fragility of the band. In this article, a new detection procedure is proposed. This new procedure takes the relationship of one site with the others into consideration and can be applied to tests of the randomness of breakpoints under either the proportional probability model (PPM) or the equiprobability model (EPM). The procedure can form a grouping structure that classifies all sites into several clusters. It is applied to identification of fragile sites for a real data set for Chinese patients with colorectal carcinoma for illustration of the proposed method. PMID- 11414568 TI - Statistical measures of DNA sequence dissimilarity under Markov chain models of base composition. AB - In molecular biology, the issue of quantifying the similarity between two biological sequences is very important. Past research has shown that word-based search tools are computationally efficient and can find some new functional similarities or dissimilarities invisible to other algorithms like FASTA. Recently, under the independent model of base composition, Wu, Burke, and Davison (1997, Biometrics 53, 1431 1439) characterized a family of word-based dissimilarity measures that defined distance between two sequences by simultaneously comparing the frequencies of all subsequences of n adjacent letters (i.e., n-words) in the two sequences. Specifically, they introduced the use of Mahalanobis distance and standardized Euclidean distance into the study of DNA sequence dissimilarity. They showed that both distances had better sensitivity and selectivity than the commonly used Euclidean distance. The purpose of this article is to extend Mahalanobis and standardized Euclidean distances to Markov chain models of base composition. In addition, a new dissimilarity measure based on Kullback-Leibler discrepancy between frequencies of all n-words in the two sequences is introduced. Applications to real data demonstrate that Kullback-Leibler discrepancy gives a better performance than Euclidean distance. Moreover, under a Markov chain model of order kQ for base composition, where kQ is the estimated order based on the query sequence, standardized Euclidean distance performs very well. Under such a model, it performs as well as Mahalanobis distance and better than Kullback-Leibler discrepancy and Euclidean distance. Since standardized Euclidean distance is drastically faster to compute than Mahalanobis distance, in a usual workstation/PC computing environment, the use of standardized Euclidean distance under the Markov chain model of order kQ of base composition is generally recommended. However, if the user is very concerned with computational efficiency, then the use of Kullback-Leibler discrepancy, which can be computed as fast as Euclidean distance, is recommended. This can significantly enhance the current technology in comparing large datasets of DNA sequences. PMID- 11414569 TI - A regression modeling approach for describing patterns of HIV genetic variation. AB - We introduce a novel approach for describing patterns of HIV genetic variation using regression modeling techniques. Parameters are defined for describing genetic variation within and between viral populations by generalizing Simpson's index of diversity. Regression models are specified for these variation parameters and the generalized estimating equation framework is used for estimating both the regression parameters and their corresponding variances. Conditions are described under which the usual asymptotic approximations to the distribution of the estimators are met. This approach provides a formal statistical framework for testing hypotheses regarding the changing patterns of HIV genetic variation over time within an infected patient. The application of these methods for testing biologically relevant hypotheses concerning HIV genetic variation is demonstrated in an example using sequence data from a subset of patients from the Multicenter AIDS Cohort Study. PMID- 11414570 TI - Catch estimation with restricted randomization in the effort survey. AB - One common method for estimating total catch is to multiply an estimate for CPUE, the catch per unit effort, by an estimate of total effort obtained from an independent second survey. In general, estimating total effort requires that sample times are chosen at random over the full fishing period; however, in practice, this may not always be possible and the usual estimator may be severely biased. Such a restriction in randomization is likely when aircraft are used to make instantaneous counts of fishing activity. This article proposes alternate estimators for use with both access and roving designs in conjunction with effort surveys for which sample times are not random. Ratio type estimators based on activity counts are developed under various scenarios and their performance examined under simulation. In addition, optimizing strategies for use with multiple activity counts are explored. Finally, data from an in-river gill net fishery on the Fraser River is used to illustrate these results. PMID- 11414571 TI - Abalone I: analyzing mark-recapture-recovery data incorporating growth and delayed recovery. AB - Abalone are semimobile marine gastropods that form the basis of Australia's second most valuable fishery. A site off the coast of Port Arthur, Tasmania, was visited on six occasions. On each occasion, any unmarked live abalone found were marked with a unique identification number and were recorded. Any previously marked abalone found had its identification number and whether or not it was still alive recorded. This results in integrated mark-recapture-recovery data, as in Catchpole et al. (1998, Biometrics 54, 33-46). During the study period, abalone grew in size, and we model the survival of individuals as a function of their size, estimated from a fitted growth curve. The shells of dead animals are long lasting, and we extend existing methodology to allow for the possibility that an animal found dead may have been dead but overlooked for several visits. PMID- 11414572 TI - Unconditional exact tests for equivalence or noninferiority for paired binary endpoints. AB - Problems of establishing equivalence or noninferiority between two medical diagnostic procedures involve comparisons of the response rates between correlated proportions. When the sample size is small, the asymptotic tests may not be reliable. This article proposes an unconditional exact test procedure to assess equivalence or noninferiority. Two statistics, a sample-based test statistic and a restricted maximum likelihood estimation (RMLE)-based test statistic, to define the rejection region of the exact test are considered. We show the p-value of the proposed unconditional exact tests can be attained at the boundary point of the null hypothesis. Assessment of equivalence is often based on a comparison of the confidence limits with the equivalence limits. We also derive the unconditional exact confidence intervals on the difference of the two proportion means for the two test statistics. A typical data set of comparing two diagnostic procedures is analyzed using the proposed unconditional exact and asymptotic methods. The p-value from the unconditional exact tests is generally larger than the p-value from the asymptotic tests. In other words, an exact confidence interval is generally wider than the confidence interval obtained from an asymptotic test. PMID- 11414573 TI - Unbiased confidence intervals for the odds ratio of two independent binomial samples with application to case-control data. AB - The problem of confidence interval construction for the odds ratio of two independent binomial samples is considered. Two methods of eliminating the nuisance parameter from the exact likelihood, conditioning and maximization, are described. A conditionally exact tail method exists by putting together upper and lower bounds. A shorter interval can be obtained by simultaneous consideration of both tails. We present here new methods that extend the tail and simultaneous approaches to the maximized likelihood. The methods are unbiased and applicable to case-control data, for which the odds ratio is important. The confidence interval procedures are compared unconditionally for small sample sizes in terms of their expected length and coverage probability. A Bayesian confidence interval method and a large-sample chi2 procedure are included in the comparisons. PMID- 11414574 TI - Exploring extra-binomial variation in teratology data using continuous mixtures. AB - Discrete data from animal teratology experiments are known to exhibit extra binomial variation. For example, we discuss a dominant lethal assay experiment in which male mice are exposed to various levels of radiation and are then mated to females. The response of interest is the number of resorptions out of the number of implantations. Most statistical work on analyzing such data has focused on modeling response rates as a function of dose of a suspected teratogen (radiation in this case) while accounting for the extra-binomial variability when calculating standard errors of the regression coefficients. Sometimes, however, when an unobserved genetic or exposure variable is suspected, the shape of the mixing distribution is of interest. We propose a mixture of beta-binomials (MBB) family of distributions that includes the non-parametric mixture of binomials model of Laird (1978) as a special case. The MBB family can accommodate a mixing distribution with one or more modes, and we develop a bootstrap test for multimodality. We apply the method to data from a dominant lethal teratology experiment. PMID- 11414575 TI - Similarity indices for spatial ecological data. AB - We present a method for assessing similarity between species maps of presence and absence or abundance that emphasizes global features while ignoring minor local dissimilarities. The method arranges sites into small groups, or cliques, and allows controlled changes to be made within cliques to reduce the influence of local discrepancies. Resulting measures of similarity are visually more satisfactory than traditional indices. We show that the similarity indices are useful for model selection by comparing observed spatial patterns with those predicted by different fitted models. Examples are provided for spatial distributions of oribatid mites (Acari, Oribatei), woodlarks (Lullula arborea), and red deer (Cervus elaphus). PMID- 11414576 TI - Bayesian inference for a random tessellation process. AB - This article describes an inhomogeneous Poisson point process in the plane with an intensity function based on a Dirichlet tessellation process and a method for using observations on the point process to make fully Bayesian inferences about the underlying tessellation. The method is implemented using a Markov chain Monte Carlo approach. An application to modeling the territories of clans of badgers, Meles meles, is described. PMID- 11414577 TI - Testing local independence between two point processes. AB - Dependencies between two types of points in a spatial point process can be due either to a real dependence between the two types or to the dependence on common underlying variables. We propose a global test for dependence between two point processes that is valid for a wide range of models. In contrast with previously proposed methods, it is based on a number of local test statistics, which makes it possible to map the local association between the two processes. The behavior of the test is evaluated by a simulation study. It is then applied to a vegetation pattern data set from Burkina Faso. PMID- 11414578 TI - Bayesian nonparametric modeling using mixtures of triangular distributions. AB - Nonparametric modeling is an indispensable tool in many applications and its formulation in an hierarchical Bayesian context, using the entire posterior distribution rather than particular expectations, increases its flexibility. In this article, the focus is on nonparametric estimation through a mixture of triangular distributions. The optimality of this methodology is addressed and bounds on the accuracy of this approximation are derived. Although our approach is more widely applicable, we focus for simplicity on estimation of a monotone nondecreasing regression on [0, 1] with additive error, effectively approximating the function of interest by a function having a piecewise linear derivative. Computationally accessible methods of estimation are described through an amalgamation of existing Markov chain Monte Carlo algorithms. Simulations and examples illustrate the approach. PMID- 11414579 TI - Model selection in estimating equations. AB - Model selection is a necessary step in many practical regression analyses. But for methods based on estimating equations, such as the quasi-likelihood and generalized estimating equation (GEE) approaches, there seem to be few well studied model selection techniques. In this article, we propose a new model selection criterion that minimizes the expected predictive bias (EPB) of estimating equations. A bootstrap smoothed cross-validation (BCV) estimate of EPB is presented and its performance is assessed via simulation for overdispersed generalized linear models. For illustration, the method is applied to a real data set taken from a study of the development of ewe embryos. PMID- 11414580 TI - UMPU and alternative tests for association in 2 x 2 tables. AB - The use of the uniformly most powerful among the unbiased (UMPU) test was recently suggested for the study of gametic association between two polymorphic loci as an alternative to the Fisher's exact test (Zapata and Alvarez, 1997, Annals of Human Genetics 61, 71-77). However, the proposed test is not UMPU for two-sided alternatives. In this study, we present the UMPU test, discuss criticisms against the use of randomized tests, and compare the power of several tests. We show that, in many practical cases, the use of the UMPU test is less than desirable and propose the alternative adjusted-more extreme tables (A-MET) and the equal-tails (ET) tests. We suggest that some of the general arguments against the use of randomized tests can be alleviated by a newly proposed extended p-value definition. PMID- 11414581 TI - Simple incorporation of interactions into additive models. AB - Often, the functional form of covariate effects in an additive model varies across groups defined by levels of a categorical variable. This structure represents a factor-by-curve interaction. This article presents penalized spline models that incorporate factor-by-curve interactions into additive models. A mixed model formulation for penalized splines allows for straightforward model fitting and smoothing parameter selection. We illustrate the proposed model by applying it to pollen ragweed data in which seasonal trends vary by year. PMID- 11414582 TI - Statistical inference in a two-compartment model for hematopoiesis. AB - We present a method for parameter estimation in a two-compartment hidden Markov model of the first two stages of hematopoiesis. Hematopoiesis is the specialization of stem cells into mature blood cells. As stem cells are not distinguishable in bone marrow, little is known about their behavior, although it is known that they have the ability to self-renew or to differentiate to more specialized (progenitor) cells. We observe progenitor cells in samples of bone marrow taken from hybrid cats whose cells contain a natural binary marker. With data consisting of the changing proportions of this binary marker over time from several cats, estimates for stem cell self-renewal and differentiation parameters are obtained using an estimating equations approach. PMID- 11414583 TI - Bayesian spatiotemporal inference in functional magnetic resonance imaging. AB - Mapping of the human brain by means of functional magnetic resonance imaging (fMRI) is an emerging field in cognitive and clinical neuroscience. Current techniques to detect activated areas of the brain mostly proceed in two steps. First, conventional methods of correlation, regression, and time series analysis are used to assess activation by a separate, pixelwise comparison of the fMRI signal time courses to the reference function of a presented stimulus. Spatial aspects caused by correlations between neighboring pixels are considered in a separate second step, if at all. The aim of this article is to present hierarchical Bayesian approaches that allow one to simultaneously incorporate temporal and spatial dependencies between pixels directly in the model formulation. For reasons of computational feasibility, models have to be comparatively parsimonious, without oversimplifying. We introduce parametric and semiparametric spatial and spatiotemporal models that proved appropriate and illustrate their performance applied to visual fMRI data. PMID- 11414584 TI - Efficiency gain from auxiliary data requiring additional nuisance parameters. AB - In a mark-recapture study of an animal population, live-recapture information may be supplemented by resightings from marked animals obtained throughout the period of the study and the two types of data analyzed simultaneously. The resighting data can only contribute to estimates of survival probability if they are jointly modeled with the live-recapture data and require the inclusion of additional nuisance parameters. We show that, under quite general conditions, estimates of the original parameters are estimated with the same, or improved, precision despite the inclusion of the nuisance parameters. Banding data from female goldeneye ducks (Bucephala clangula) are used as an illustration. PMID- 11414585 TI - Drawbacks to integer scoring for ordered categorical data. AB - Linear rank tests are widely used when testing for independence against stochastic order in a 2 x J contingency table with two treatments and J ordered outcome levels. For this purpose, numerical scores are assigned, possibly by default, to the J outcome levels. When the choice of scores is not apparent, integer (equally spaced) scores are often assigned. We show that this practice generally leads to unnecessarily conservative tests. The use of slightly perturbed scores will result in a less conservative and uniformly more powerful test. PMID- 11414586 TI - Analyte identification in multivariate calibration. AB - In calibration experiments, an estimated relationship between covariate information for a sample and an observed response is used to infer the covariate information for unknown samples from their responses. In some situations, this covariate information comprises a nominal variable (e.g., identity of a chemical, sex of an animal) and a real-valued variable (e.g., concentration of the chemical, age of animal). If the calibrating relationship can be estimated separately for each candidate identity, the first step in analyzing unknown samples is to correctly determine their identity. A discrimination statistic is suggested for use in this situation and its asymptotic distribution is derived. The investigation is motivated by the possibility of using multiple immunoassays in environmental monitoring to identify and quantitate contaminated samples in situations where there are several candidate pollutants that cross-react significantly to single assays. An example is given of the use of a four-antibody assay for the simultaneous monitoring of the levels in water samples of several of the commonly used triazine herbicides and their derivatives. PMID- 11414587 TI - Multiple temporal cluster detection. AB - This article proposes a simple method to determine single or multiple temporal clustering on a variable size population. By a transformation of the data set, the method based on a regression model allows consideration of a variable population size during the time of study. A model selection procedure and a resampling method are used to select the number of clusters. The results have applications in epidemiological studies of rare diseases. PMID- 11414588 TI - Inference procedures for assessing interobserver agreement among multiple raters. AB - We propose a new procedure for constructing inferences about a measure of interobserver agreement in studies involving a binary outcome and multiple raters. The proposed procedure, based on a chi-square goodness-of-fit test as applied to the correlated binomial model (Bahadur, 1961, in Studies in Item Analysis and Prediction, 158-176), is an extension of the goodness-of-fit procedure developed by Donner and Eliasziw (1992, Statistics in Medicine 11, 1511 1519) for the case of two raters. The new procedure is shown to provide confidence-interval coverage levels that are close to nominal over a wide range of parameter combinations. The procedure also provides a sample-size formula that may be used to determine the required number of subjects and raters for such studies. PMID- 11414589 TI - Estimation of a secondary parameter in a group sequential clinical trial. AB - In this article, we investigate estimation of a secondary parameter in group sequential tests. We study the model in which the secondary parameter is the mean of the normal distribution in a subgroup of the subjects. The bias of the naive secondary parameter estimator is studied. It is shown that the sampling proportions of the subgroup have a crucial effect on the bias: As the sampling proportion of the subgroup at or just before the stopping time increases, the bias of the naive subgroup parameter estimator increases as well. An unbiased estimator for the subgroup parameter and an unbiased estimator for its variance are derived. Using simulations, we compare the mean squared error of the unbiased estimator to that of the naive estimator, and we show that the differences are negligible. As an example, the methods of estimation are applied to an actual group sequential clinical trial, The Beta-Blocker Heart Attack Trial. PMID- 11414590 TI - Case-control analysis with partial knowledge of exposure misclassification probabilities. AB - Consider case control analysis with a dichotomous exposure variable that is subject to misclassification. If the classification probabilities are known, then methods are available to adjust odds-ratio estimates in light of the misclassification. We study the realistic scenario where reasonable guesses, but not exact values, are available for the classification probabilities. If the analysis proceeds by simply treating the guesses as exact, then even small discrepancies between the guesses and the actual probabilities can seriously degrade odds-ratio estimates. We show that this problem is mitigated by a Bayes analysis that incorporates uncertainty about the classification probabilities as prior information. PMID- 11414591 TI - Latent class modeling approaches for assessing diagnostic error without a gold standard: with applications to p53 immunohistochemical assays in bladder tumors. AB - Improved characterization of tumors for purposes of guiding treatment decisions for cancer patients will require that accurate and reproducible assays be developed for a variety of tumor markers. No gold standards exist for most tumor marker assays. Therefore, estimates of assay sensitivity and specificity cannot be obtained unless a latent class model-based approach is used. Our goal in this article is to estimate sensitivity and specificity for p53 immunohistochemical assays of bladder tumors using data from a reproducibility study conducted by the National Cancer Institute Bladder Tumor Marker Network. We review latent class modeling approaches proposed by previous authors, and we find that many of these approaches impose assumptions about specimen heterogeneity that are not consistent with the biology of bladder tumors. We present flexible mixture model alternatives that are biologically plausible for our example, and we use them to estimate sensitivity and specificity for our p53 assay example. These mixture models are shown to offer an improvement over other methods in a variety of settings, but we caution that, in general, care must be taken in applying latent class models. PMID- 11414592 TI - Likelihood-based modeling and analysis of data underdispersed relative to the Poisson distribution. AB - By using a generalization of the Poisson process, distributions can be constructed that show appropriate amounts of underdispersion relative to the Poisson distribution that may be apparent from observed data. These are then used to examine the differences between the distributions of numbers of fetal implants in mice corresponding to different doses of the herbicide 2,4,5-T. PMID- 11414593 TI - Controlling test size while gaining the benefits of an internal pilot design. AB - To compensate for a power analysis based on a poor estimate of variance, internal pilot designs use some fraction of the planned observations to reestimate error variance and modify the final sample size. Ignoring the randomness of the final sample size may bias the final variance estimate and inflate test size. We propose and evaluate three different tests that control test size for an internal pilot in a general linear univariate model with fixed predictors and Gaussian errors. Test 1 uses the first sample plus those observations guaranteed to be collected in the second sample for the final variance estimate. Test 2 depends mostly on the second sample for the final variance estimate. Test 3 uses the unadjusted variance estimate and modifies the critical value to bound test size. We also examine three sample-size modification rules. Only test 2 can control conditional test size, align with a modification rule, and provide simple power calculations. We recommend it if the minimum second (incremental) sample is at least moderate (perhaps 20). Otherwise, the bounding test appears to have the highest power in small samples. Reanalyzing published data highlights some advantages and disadvantages of the various tests. PMID- 11414594 TI - Relating amino acid sequence to phenotype: analysis of peptide-binding data. AB - We illustrate data analytic concerns that arise in the context of relating genotype, as represented by amino acid sequence, to phenotypes (outcomes). The present application examines whether peptides that bind to a particular major histocompatibility complex (MHC) class I molecule have characteristic amino acid sequences. However, the concerns identified and addressed are considerably more general. It is recognized that simple rules for predicting binding based solely on preferences for specific amino acids in certain (anchor) positions of the peptide's amino acid sequence are generally inadequate and that binding is potentially influenced by all sequence positions as well as between-position interactions. The desire to elucidate these more complex prediction rules has spawned various modeling attempts, the shortcomings of which provide motivation for the methods adopted here. Because of (i) this need to model between-position interactions, (ii) amino acids constituting a highly (20) multilevel unordered categorical covariate, and (iii) there frequently being numerous such covariates (i.e., positions) comprising the sequence, standard regression/classification techniques are problematic due to the proliferation of indicator variables required for encoding the sequence position covariates and attendant interactions. These difficulties have led to analyses based on (continuous) properties (e.g., molecular weights) of the amino acids. However, there is potential information loss in such an approach if the properties used are incomplete and/or do not capture the mechanism underlying association with the phenotype. Here we demonstrate that handling unordered categorical covariates with numerous levels and accompanying interactions can be done effectively using classification trees and recently devised bump-hunting methods. We further tackle the question of whether observed associations are attributable to amino acid properties as well as addressing the assessment and implications of between position covariation. PMID- 11414595 TI - Regret at the use of the word "biometrics". PMID- 11414596 TI - Fatal pediatric lead poisoning--New Hamphshire, 2000. AB - Fatal pediatric lead poisoning is rare in the United States because of multiple public health measures that have reduced blood lead levels (BLLs) in the population. However, the risk for elevated BLLs among children remains high in some neighborhoods and populations, including children living in older housing with deteriorated leaded paint. This report describes the investigation of the first reported death of a child from lead poisoning since 1990 (1). The investigation implicated leaded paint and dust in a home environment as the most likely source of the poisoning. Lead poisoning can be prevented by correcting lead hazards, especially in older housing, and by screening children at risk according to established guidelines (2). PMID- 11414597 TI - Evaluation of sexually transmitted disease control practices for male patients with urethritis at a large group practice affiliated with a managed care organization--Massachusetts, 1995-1997. AB - Effective management for sexually transmitted diseases (STDs) depends on appropriate testing, treatment, partner management, and complete and timely reporting of positive STD tests (1). Testing can ensure appropriate treatment of initial or recurrent infections and identification of drug-resistant pathogens, appropriate treatment can reduce risk for complications and development of drug resistance, and complete and timely reporting of positive test results by laboratories and STD cases by health-care providers to health departments can facilitate rapid sex partner notification and outbreak detection. By 1998, private providers, including those affiliated with commercial or Medicaid managed care organizations (MCOs) (2,3) were caring for approximately 70% of persons with chlamydia and 55% of persons with gonorrhea. To assess the quality of STD care at a MCO-affiliated multisite facility, the testing, treatment, and reporting practices of gonorrhea- and chlamydia-associated urethritis in male patients were evaluated. This report summarizes the evaluation, which indicated that the providers tested most men with urethritis symptoms, prescribed CDC-recommended therapy to all patients, and reported most laboratory-confirmed chlamydia and gonorrhea cases of urethritis to the state health department. Several interventions introduced at this large group practice may have encouraged these favorable STD practices. PMID- 11414598 TI - Racial disparities in median age at death of persons with Down syndrome--United States, 1968-1997. AB - Down syndrome (DS) is the most common identified cause of mental retardation in the United States (1). The prevalence (approximately one in 800 live-born infants) is similar among all racial groups (2). Survival for the first year of life for infants with DS has improved dramatically during the last 50 years, from <50% in a 1942-1952 birth cohort (3) to 91% in a 1980-1996 cohort (4). Most studies of survival in persons with DS have focused on white populations, and little information is available about possible disparities among racial groups. To investigate changes in the age at death among persons with DS by race, CDC analyzed data from multiple-cause mortality files (MCMF) for 1968-1997. This report summarizes the results of the analysis, which indicate that the median age at death of persons with DS increased substantially during this period, but this increase was much greater for whites than for blacks or other races. Identification of the causes for this racial disparity may permit development of strategies to improve the survival of persons with DS, especially those who are black or of other racial groups. PMID- 11414599 TI - Update: influenza activity--United States and worldwide, 2000-01 season, and composition of the 2001-02 influenza vaccine. AB - The 2000-01 influenza season was mild in the United States and was the first season since 1995-96 that was not predominated by A (H3N2) viruses. Influenza A (H1N1) viruses predominated in the United States. In some regions, however, influenza B viruses were reported more frequently than influenza A viruses. Worldwide, influenza A (H1N1) and B viruses also predominated. This report summarizes U.S.* and worldwide influenza activity during the 2000-01 influenza season and describes the composition of the 2001-02 influenza vaccine. PMID- 11414600 TI - The elephant trunk technique for type A dissection. AB - A modified elephant trunk technique that was used to treat acute type A dissection is described. This technique prevents leaks at the distal anastomosis and facilitates surgery on the aneurysmatic downstream aorta following total arch replacement. In addition, it allows closure of the primary intimal tear in patients with DeBakey type III retrograde dissection. PMID- 11414601 TI - Anesthesia for heart or single or double lung transplantation in the adult patient. AB - Providing an anesthetic for patients undergoing heart or a single or double lung transplantation may represent a challenge even to the most experienced anesthesiologist. Patients with end-stage cardiac dysfunction have an impaired response to beta-agonist due to receptor downregulation. These patient will have isolated left ventricular dysfunction secondary to ischemic heart disease or present with biventricular failure with or without significant pulmonary hypertension. Increasingly, more patients have undergone prior major cardiac procedures and are at risk for significant perioperative bleeding. Patients undergoing single or double lung are particularly challenging because most of these procedures are performed without the aid of cardiopulmonary bypass. The anesthesiologist must be proficient at the management of one-lung ventilation techniques and have a rational physiologic approach to the management of intraoperative hypoxemia and auto-PEEP. PMID- 11414602 TI - PEEP therapy for patients with pleurotomy during coronary artery bypass grafting. AB - Severe pulmonary oxygenation impairment resulting from peripheral lung atelectasis occurred in some patients with pleurotomy during the harvest of the internal mammary artery graft followed by coronary artery bypass grafting (CABG). We studied the efficacy of intraoperative positive end-expiratory airway pressure (PEEP) therapy for the prevention of postoperative pulmonary oxygenation impairment. A total of 66 patients with solitary CABG procedure were included in this study. The pleural cavity was intraoperatively opened in 44 patients and not opened in 22. PEEP therapy was not used in any patient before May 1996 (referred to herein as the former period) and was used more recently in eight patients with pleurotopmy (referred to herein as the latter period). PEEP was initiated immediately after pleurotomy during the harvest of the internal mammary artery graft. Without PEEP therapy, values of PaO2, A-aDO2, and respiratory index (RI) were worse in patients with pleurotomy than in those without pleurotomy. Meanwhile, there were no major differences in these values between patients with or without pleurotomy after the induction of PEEP therapy. Respiratory insufficiency (A-aDO2 > 400 mmHg and RI > 1.5) was detected in six patients with pleurotomy in the former period. Three of these six patients required over 1 week of long-term mechanical respiratory support. No respiratory insufficiency occurred in patients of the latter period. In conclusion, PEEP therapy, which is initiated just after pleurotomy, may prevent oxygen impairment and pulmonary atelectasis after extracorporeal circulation (ECC) and is recommended for patients with pleurotomy, especially for patients with preoperative low respiratory function. PMID- 11414603 TI - Partial left ventriculectomy: overall and late results in 44 class IV patients with 4-year follow-up. AB - BACKGROUND: This study reports long-term results of partial left ventriculectomy (PLV). METHODS: Forty-four patients with dilated cardiomyopathy were operated on in a 4-year study. Echocardiograms, catheterization, and stress tests with oxygen consumption (VO2) were performed. RESULTS: The survivors' preoperative ejection fractions of 22.1% +/- 4.9% improved to 30.9% +/- 9.4%, left ventricular (LV) end diastolic diameter decreased from 79.4 +/- 9.3 mm to 61.9 +/- 8.2 mm, and maximum VO2 consumption improved from 8.8 +/- 3.9 mL/kg per minute to 15.8 +/- 6.1 mL/kg per minute at 22.6 months. These data also showed improvements in nonsurviving patients, according to the last evaluation before death. Seven of 12 survivors (58.3%) were in New York Heart Association (NYHA) I and II in December 1998. Twelve patients had elevated pulmonary vascular resistance (PVR) contraindicating heart transplant. In five patients the PVR returned to normal and one high-PVR patient was transplanted at the 16th postoperative month. Survival was 56.8%, 47.7%, 38.4%, and 35.9%, respectively, at 3, 6, 12, and 18 months, with a tendency to stabilize at 32.7% thereafter. Arrhythmias and heart failure were the main causes of death. CONCLUSIONS: In spite of improvement of ventricular function and quality of life of the survivors, high mortality is a limiting factor. PLV can be indicated as a bridge to heart transplantation in high-PVR patients or if ventricular assist devices or donor hearts are not available. PMID- 11414604 TI - Determinants of early mortality and neurological morbidity in aortic operations performed under circulatory arrest. AB - OBJECTIVE: Aneurysms and dissections of the thoracic aorta continue to present a surgical challenge and their incidence is increasing in recent years. The mortality rate of surgical treatment is still higher than those of other cardiovascular operations. Neurological injury is the most feared complication resulting from repair of these lesions. This study aims to determine the factors that influence the neurological outcome and mortality after thoracic aortic operations. METHODS: During the period from November 1993 through May 1999, 144 patients were operated on for conditions involving the ascending aorta and/or aortic arch. Ninety-five (66.0%) were operated for aortic dissection and 49 (34.0%) were for aortic aneurysms. Sixty-two patients (43.1%) had replacement of ascending aorta with distal open technique; 82 patients (56.9%) had hemiarch or total arch replacement or repair of the distal arch. RESULTS: Twenty-seven (18.7%) early deaths occurred. New stroke occurred in two patients (1.4%) and temporary neurological dysfunction in nine patients (6.3%). Deep hypothermic circulatory arrest with retrograde cerebral perfusion was used in all patients. On multivariate logistic regression analysis, risk factors for mortality were chronic renal failure, preoperative organ malperfusion, rupture, total circulatory arrest time > 60 minutes, postoperative acute renal failure, postoperative low cardiac output, sepsis, and multiple organ failure. Risk factors for neurological morbidity were preoperative chronic renal failure, preoperative hemodynamic instability, postoperative low cardiac output, and pulmonary complications. CONCLUSIONS: Hypothermic circulatory arrest with retrograde cerebral perfusion was not an independent predictor of neurological morbidity on multivariate analysis, even if the arrest period was more than 60 minutes. Lengths of circulatory arrest periods and clinical presentations of the patients are important determinants of mortality. PMID- 11414605 TI - Heparin-coated cardiopulmonary bypass circuit: clinical effects in pediatric cardiac surgery. AB - OBJECTIVES: Heparin-coated cardiopulmonary bypass (CPB) circuits have been reported to reduce complement activation and the inflammatory response associated with CPB. We retrospectively compared patients utilizing heparin-coated perfusion circuits with those using noncoated circuits to determine the clinical effects of the different circuits in pediatric cardiac surgery. METHODS: Between July 1995 and July 1997, 203 patients weighing < 10 kg underwent cardiac surgery, 153 patients using heparin-coated bypass circuits and 50 patients using noncoated circuits. The 50 patients operated on with the noncoated circuit (Group N) were matched to 100 patients operated on with coated circuits (Group H) in age, weight, and type of procedure. Urine output during bypass, blood products used after bypass, postoperative ventilation days, hospital stay, morbidity, and mortality were compared between these groups. RESULTS: Body weight, perfusion time, and procedure time were not different between the two groups. Urine output during bypass was notably greater in Group H than in Group N (11.3 +/- 10.5 mL/kg per hour vs 4.8 +/- 3.1 mL/kg per hour, respectively, p < 0.0001). Postoperative mechanical ventilation markedly decreased in Group H (Group H vs N = 2.8 +/- 2.7 days vs 5.1 +/- 7.5 days, respectively, p < 0.05). Red blood cell usage, hospital stay, morbidity, and mortality were not statistically different, although there was a tendency toward decreased transfusion of red cell and platelets in Group H (Group H vs N = 61.2 +/- 121.1 mL/kg vs 102.0 +/- 176.7 mL/kg, respectively, in red cell, p = 0.15; and Group H vs N = 7.9 +/- 13.7 mL/kg vs 13.2 +/- 24.5 mL/kg, respectively, in platelets, p = 0.16). CONCLUSIONS: Patients operated on with the use of heparin-coated circuits had increased urine output during bypass and required less time postoperatively on the ventilator. These results suggest a reduction in the acute inflammatory response, capillary leakage, and overall systemic edema. We now routinely use coated circuits on all pediatric pump cases. PMID- 11414606 TI - Multistep approach to electrical stimulation after cardiomyoplasty. AB - Still a controversial procedure, cardiomyoplasty (CMP) improves the failing heart's ability to contract by using a latissimus dorsi muscle (LDM), but to date, hemodynamic results correlate poorly with clinical improvement. The following two causes for apprehension bar attempting to change the conventional electrical stimulation (ES) protocol to improve CMP results: (1) fear of beginning ES for LDM-assisted contraction immediately postmobilization and CMP and (2) fear of stopping or slowing ES during sleep periods. METHODS: In ten different experimental series, I used animal models of CMP to determine how to apply ES to newly mobilized LDM, how to begin partial cardiac assist immediately post-CMP, and how to suspend ES for 12 hours daily. RESULTS: From my experimental series I noted the following three results. (1) Different ES regimens applied 1 hour postmobilization changed the contractile force (CF). After a 30-minute fatigue test, CF decreased by 34% +/- 3% with continuous ES at 30 contractions per minute (cpm), by 23% +/- 2% with continuous ES at 15 cpm, by 25% +/- 5% with ES in a work-rest regimen at 30 cpm, and by 8% +/- 3% with ES in a work-rest regimen at 15 cpm. (2) Imitation of partial cardiac assist immediately postmobilization slightly decreased CF. Sixteen days postmobilization, during a 30-minute fatigue test in animals in which ES had been started immediately after mobilization, CF decreased by only 6% +/- 3% and did not change when ES was combined with imitation of cardiac assist for 30 minutes twice daily (work-rest regimen). (3) ES cessation for 24 hours daily or 12 hours daily in CMP model created no difference in ejection fraction (EF) with ES (54% +/- 4% vs 53% +/- 5%, respectively (or in left ventricular end-diastolic volume (LVEDV, 234.3 ml +/ 1.0 ml vs 24.8 mL +/- 0.6 mL, respectively) or in LV end-systolic volume (LVESV; 12.1 mL +/- 0.7 mL vs 12.8 mL +/- 0.7 mL, respectively). CONCLUSION: For improving angiographic potential in the LDM, ES can be started safely immediately post-CMP at 15 cpm (a 1:4 or 1:5 regimen) and single impulses per burst. For partial cardiac assist and for improving LDM performance, cardiac assist can be used for 30 minutes twice daily immediately post-CMP. To rest the muscle and save it from overuse, muscle contraction can be either stopped or slowed down during hours of sleep. PMID- 11414607 TI - Ventricular aneurysm repair: a new approach. AB - Repair of chronic left ventricular aneurysm or acute rupture of the heart after myocardial infarction is associated with technical difficulties and major morbidity and mortality. We describe a new endoventricular repair for both conditions. The repair consists of externally covering a Duran ring with Dacron and internally lining it with autologous pericardium. The modified ring is then sewn into the neck of the lesion. The technique is rapid, simple, and hemostatic. After repair of the chronic aneurysm, ventricular hemodynamics are improved during both diastole and systole. We have performed this technique in two patients with chronic aneurysm and one with subacute rupture. PMID- 11414608 TI - Is left main coronary artery stenosis a risk factor for early mortality in coronary artery surgery? AB - It is accepted universally that the treatment of critical left main coronary artery (LMCA) stenosis is surgical revascularization. This study was designed to evaluate critical LMCA stenosis as a risk factor in coronary artery bypass surgery. We compared the surgical results of 760 patients with critical LMCA disease, including 58 cases who were operated under emergency conditions (LMCA em) and 702 patients who were operated electively (LMCA-el), with randomly chosen 707 coronary bypass patients (CONT-el) without LMCA disease, but who had double- or triple-vessel disease. Another group of patients (n = 99) who were operated on under emergency conditions (CONT-em) but without LMCA disease were also compared with the corresponding groups. The mortality of LMCA-em group and CONT-em group was markedly higher from the other two groups. Univariate analysis revealed that female gender, older age, presence of diabetes mellitus, poor left ventricular function, and the presence of unstable angina were major risk factors for fatal outcome in LMCA-el and CONT-el groups. Age was also a risk factor in LMCA-em group, as well as unstable angina pectoris. The coexistence of critical right coronary artery disease did not affect the early outcome in both groups with LMCA lesions. In the multivariate analysis, critical LMCA disease was not a risk factor for mortality. Logistic regression analysis revealed diabetes [odds ratio (OR): 3.66], poor left ventricular function (higher left ventricle end-diastolic pressure, OR: 1.08), and emergent operations (OR: 5.09) were risk factors for early mortality. Patients with LMCA disease should have surgery promptly for favorable results, because emergency conditions have higher mortality rates. PMID- 11414609 TI - Unusual recurrence of hydatid cysts of the heart: report of two cases and review of the clinical and surgical aspects of the disease. AB - In cardiac echinococcosis, a hydatid cyst most frequently forms either solely in the heart or in the pericardium, but there are several reports of cysts forming in the liver or lung or in both. In two cases reported here, both patients developed cysts in new sites after one or more previous surgeries for hydatid cyst removal. In Case 1, the patient first underwent spleenectomy and resection of multiple cysts with no evidence of a cyst in the heart; 3 years later, there was no sign of Echinococcus in the liver, but a large inframyocardial cyst had damaged the left ventricle. In Case 2, the patient first underwent surgery to remove cysts from the pericardium, 2 years later from the anterior wall of the left ventricle, and, finally, 8 months after this second operation, from the left atrium also with no evidence of cyst formation anywhere else in the heart at the time of surgery. These cases emphasize the need for thorough and frequent reevaluation to detect new hydatid cyst formation in the heart and elsewhere caused by the Echinococcus organism. PMID- 11414610 TI - Downstream DNA sequences are required to modulate Pvlea-18 gene expression in response to dehydration. AB - We have previously shown that mRNA and protein encoded by the Pvlea-18 gene from Phaseolus vulgaris L., a member of a new family of late embryogenesis-abundant (LEA) proteins, accumulate in dark-grown bean seedlings not only in response to water deficit but also during optimal irrigation. In this work, we studied Pvlea 18 gene transcriptional regulation by using transgenic Arabidopsis thaliana plants containing a chimeric gene consisting of the Pvlea-18 promoter region and the 3'-nos terminator fused to the GUS gene-coding region. We demonstrate that the chimeric gene is active during Arabidopsis normal development under well irrigated conditions, and that it is further induced in response to ABA and dehydration treatments. Replacing the 3'-nos terminator with the Pvlea-18 3' region led to an additional increase in expression during development and in response to dehydration, but not in response to exogenous ABA. These results reveal an enhancer effect of the Pvlea-18 3' region, which showed to be higher specifically under dehydration. The small decrease in Pvlea-18 promoter expression observed when transgenic plants treated with fluridone (an ABA biosynthesis inhibitor) were subjected to dehydration suggests that the Pvlea-18 gene dehydration response is predominantly ABA-independent. Finally, we present evidence indicating that Pvlea-18 gene expression is negatively regulated during etiolated growth, particularly in roots, in contrast to the expression pattern observed during normal development. PMID- 11414611 TI - Molecular identification and characterization of the Arabidopsis AtADF1, AtADFS and AtADF6 genes. AB - Actin depolymerizing factor (ADF) is a key regulator of the organization of the actin cytoskeleton during various cellular activities. We found that ADF genes in Arabidopsis form a large family consisting of at least nine members, four of which were cloned and sequenced in this study. Comparison of genomic and cDNA sequences showed that the AtADF1, AtADF5, and AtADF6 genes all contain two introns at conserved positions. Analysis of transgenic Arabidopsis plants carrying promoter-GUS fusion constructs revealed that AtADF1 and AtADF6 are expressed in the vascular tissues of all organs, whereas expression of AtADF5 is restricted to the root tip meristem. GFP-AtADFI, GFP-AtADF5, and GFP-AtADF6 fusion proteins were found to bind to actin filaments in vivo, and to reorganize the actin cytoskeleton when transiently expressed in plant cells. PMID- 11414612 TI - Molecular cloning and characterization of cDNA encoding cardosin B, an aspartic proteinase accumulating extracellularly in the transmitting tissue of Cynara cardunculus L. AB - Cardosins A and B are related aspartic proteinases from the pistils of Cynara cardunculus L., whose milk-clotting activity has been exploited for the manufacture of cheese. Here we report the cloning of cardosin B cDNA and its organ, tissue and cytological localization. The cDNA-derived amino acid sequence has 73% similarity with that of cardosin A and displays several distinguishing features. Cardosin B mRNA was detected in young inflorescences but not in pistils of fully opened inflorescences, indicating that its expression is developmentally regulated. The proteinase, however, accumulates in the pistil until the later stages of floral development. Immunocytochemistry with a monospecific antibody localized cardosin B to the cell wall and extracellular matrix of the floral transmitting tissue. The location of cardosin B in the pistil is therefore clearly different from that of cardosin A, which was found at protein storage vacuoles of the stigmatic papillae and has been suggested to be involved in RGD mediated proteolytic mechanisms. In view of these results the possible functions of cardosin B in the transmitting tissue are discussed. PMID- 11414613 TI - A cDNA from grapevine (Vitis vinifera L.), which shows homology to AGAMOUS and SHATTERPROOF, is not only expressed in flowers but also throughout berry development. AB - An AGAMOUS/SHATTERPROOF homologue (Vvmads1) was isolated from grapevine by differential display between berry and leaf mRNA. The predicted protein sequence of the full-length clone shows a high degree of homology to PLENA (77% identity) and to SHP1 and SHP2 (75% and 74% identity respectively), and is grouped with AGAMOUS/PLENA homologues when the conserved MADS and K domains are compared. Vvmads1 is expressed only in the later stages of flower development and throughout berry development, although expression is reduced after ripening commenced. When Vvmads1 was over-expressed in tobacco, the resulting plants display altered morphologies in the outer two floral whorls. In the most extreme cases, the inner whorls were surrounded by a carpelloid structure created by the modified sepals. Within these sepals were petals which had been split into sections and which were attached at the base of the flower by structures with the appearance of filaments. The results of this study suggest that Vvmads1 has a regulatory role in flower development before fertilisation and a role in fruit development after fertilisation. PMID- 11414614 TI - Regulation of loblolly pine (Pinus taeda L.) arginase in developing seedling tissue during germination and post-germinative growth. AB - After seed germination, hydrolysis of storage proteins provides a nitrogen source for the developing seedling. In conifers the majority of these reserves are located in the living haploid megagametophyte tissue. In the developing loblolly pine (Pinus taeda L.) seedling an influx of free amino acids from the megagametophyte accompanies germination and early seedling growth. The major component of this amino acid pool is arginine, which is transported rapidly and efficiently to the seedling without prior conversion. This arginine accounts for nearly half of the total nitrogen entering the cotyledons and is likely a defining factor in early seedling nitrogen metabolism. In the seedling, the enzyme arginase is responsible for liberating nitrogen, in the form of ornithine and urea, from free arginine supplied by the megagametophyte. In this report we investigate how the seedling uses arginase to cope with the large arginine influx. As part of this work we have cloned an arginase cDNA from a loblolly pine expression library. Analysis of enzyme activity data, accumulation of arginase protein and mRNA abundance indicates that increased arginase activity after seed germination is due to de novo synthesis of the enzyme. Our results suggest that arginase is primarily regulated at the RNA level during loblolly pine seed germination and post-germinative growth. PMID- 11414615 TI - Expression and activity of cell-wall-degrading enzymes in the latex of opium poppy, Papaver somniferum L. AB - The alkaloid-rich latex of the opium poppy, Papaver somniferum L., is valued as a source of pharmaceuticals including thebaine, codeine, and morphine, but is also harvested for heroin production. The poppy laticifer system develops through the gradual disappearance of the common walls between differentiating laticifer elements throughout the plant. Gene homologues for cell-wall-degrading enzymes were found during random sequencing of an opium poppy latex cDNA library. RNA gel blot analysis of cellulase, polygalacturonase beta-subunit, 1,3-beta-glucanase, and xyloglucan endotransglycosylase homologues showed their expression was not limited to laticifers. In contrast, poppy gene homologues to pectin methylesterase (PME), pectin acetylesterase (PAE) and pectate lyase (PL) where all highly expressed and latex-specific. Enzyme assays confirmed the presence of PME, PAE, and PL activities in latex serum. The abundance of transcripts encoding pectin-degrading enzymes in latex suggests that these enzymes may play an important role in laticifer development. PMID- 11414616 TI - Functional analysis of cis-regulatory elements within the promoter of the tobacco late pollen gene g10. AB - The tobacco gene g10 is preferentially and maximally expressed in mature pollen, shows homology to pectate lyases, and is the putative homologue of the tomato gene lat56. Analysis of regulatory elements within the g10 promoter was carried out to verify the importance of putative regulatory sequence motifs. Analysis of transgenic plants showed that 1190 bp of g10 5' sequence directed preferential expression of GUS in pollen, with bimodal peaks of expression just before and during pollen mitosis I, and in mature anthers. This was confirmed by northern analysis of native g10 transcripts in isolated spores. Transient expression analysis defined the minimal g10 promoter region capable of directing expression in pollen as -86 to +217. Three upstream regions within -427 bp modulate the expression from g10. Gain-of-function analyses showed that the region from -106 to -53 could enhance pollen-specific expression of a minimal CaMV 35S promoter. These analyses further showed that sequences upstream of -86 modulate expression in pollen, but are not essential for preferential pollen expression. The function of a conserved GTGA motif shared between the tobacco g10 and tomato lat56 promoters was demonstrated in g10. Thus, further functional evidence is provided for the conservation of mechanisms for the regulation of late pollen genes across species. PMID- 11414617 TI - Lectin and lectin-related proteins in lima bean (Phaseolus lunatus L.) seeds: biochemical and evolutionary studies. AB - Lectin-related polypeptides are a class of defence proteins found in seeds of Phaseolus species. In Lima bean (P. lunatus), these proteins and their genes have been well characterized in the Andean morphotype, which represents one of the two gene pools of this species. To study the molecular evolution of the lectin family in Lima bean we characterized the polypeptides belonging to this multigene family and cloned the genes belonging to the Mesoamerican gene pool. The latter gene pool contains components similar to those of the Andean pool, namely: an amylase inhibitor-like (AIL), an arcelin-like (ARL) lectin and the less abundant Lima bean lectin (LBL). These proteins originate from an ancestor gene of the lectin type which duplicated to yield the lectin gene and the progenitor of ARL and AIL. In this species. ARL represents an evolutionary intermediate form that precedes AIL. Phylogenetic analysis supports an Andean origin for Lima bean. The molecular evolutionary studies were extended to the genes of common bean and demonstrated that true lectin genes and the ancestor of lectin-related genes are the result of a duplication event that occurred before speciation. Lima and common bean followed different evolutionary pathways and in the latter species a second duplication event occurred that gave rise, in Mesoamerican wild genotypes, to arcelin genes. PMID- 11414618 TI - A constitutive cystatin-encoding gene from barley (Icy) responds differentially to abiotic stimuli. AB - A barley cDNA clone encoding a cysteine proteinase inhibitor was characterized. The deduced amino acid sequence of this barley cystatin (Hv-CPI) contains the motif QXVXG conserved among members of the cystatin superfamily. The gene (Icy), located on chromosome 2, was expressed in embryos, developing endosperms, leaves and roots as assessed by northern blot analysis. Western blot analysis detected a slightly retarded band in leaves that was not present in roots or seeds. In these two organs a more precise location of Hv-CPI was done by immuno-histochemical analysis, with polyclonal antibodies raised against the recombinant CPI protein expressed in Escherichia coli. This protein efficiently inhibited papain (Ki 2.0 x 10(-8) M) and ficin (Ki 2.2 x 10(-8) M) and, to a lesser extent, chymopapain (Ki 1.6 x 10(-7) M) and was inactive against bromelain. The Icy mRNA expression in vegetative tissues increased in response to anaerobiosis, dark and cold shock (6 degrees C). PMID- 11414619 TI - Substrate specificity and antifungal activity of recombinant tobacco class I chitinases. AB - Endochitinases contribute to the defence response of plants against chitin containing pathogens. The vacuolar class I chitinases consist of an N-terminal cysteine-rich domain (CRD) linked by a glycine-threonine-rich spacer with 4 hydroxylated prolyl residues to the catalytic domain. We examined the functional role of the CRD and spacer region in class I chitinases by comparing wild-type chitinase A (CHN A) of Nicotiana tabacum with informative recombinant forms. The chitinases were expressed in transgenic N. sylvestris plants, purified to near homogeneity, and their structures confirmed by mass spectrometry and partial sequencing. The enzymes were tested for their substrate preference towards chitin, lipo-chitooligosaccharide Nod factors of Rhizobium, and bacterial peptidoglycans (lysozyme activity) as well as for their capacity to inhibit hyphal growth of Trichoderma viride. Deletion of the CRD and spacer alone or in combination resulted in a modest <50% reduction of hydrolytic activity relative to CHN A using colloidal chitin or M. lysodeikticus walls as substrates; whereas, antifungal activity was reduced by up to 80%. Relative to CHN A, a variant with two spacers in tandem, which binds chitin, showed very low hydrolytic activity towards chitin and Nod factors, but comparable lysozyme activity and enhanced antifungal activity. Neither hydrolytic activity, substrate specificity nor antifungal activity were strictly correlated with the CRD-mediated capacity to bind chitin. This suggests that the presence of the chitin-binding domain does not have a major influence on the functions of CHN A examined. Moreover, the results with the tandem-spacer variant raise the possibility that substantial chitinolytic activity is not essential for inhibition of T. viride growth by CHN A. PMID- 11414620 TI - An alternative method for the release criteria and calculation of the total dose equivalent to another individual from a patient treated with a therapeutic dose of 131I. AB - The Nuclear Regulatory Commission's Regulatory Guide 8.39, "Release of Patients Administered Radioactive Materials," permits licensees to authorize the release of patients treated with 131I [(T 1/2)p = 8.04 d] if the total dose equivalent to another individual from the released individual is not likely to exceed 5 millisieverts (0.5 rem). This paper guides the reader through the derivation of an easy-to-use formula that demonstrates how to calculate the total dose equivalent, using patient-specific parameters that an individual may receive from a patient who has been administered a therapeutic dose of 131I, as compared to the NRC's Reg. Guide 8.39 (see Appendix A). Patient-specific parameters include the individual thyroidal uptake, the patient's physical dimensions, attenuation of radiation by body tissues, buildup of secondary photons, whole body and thyroid retention functions as well as the distance and time the patient is around others. This paper also demonstrates how to calculate an inpatient's discharge time, so that the patient will not expose other individuals above the desired total dose equivalent set by the Radiation Safety Officer (RSO). PMID- 11414621 TI - Evaluation of eight decision rules for low-level radioactivity counting. AB - In low-level radioactivity measurements, it is often important to decide whether a measurement differs from background. A traditional formula for decision level (DL) is given in numerous sources, including the recent ANSI/HPS N13.30-1996, Performance Criteria for Radiobioassay and the Multi-Agency Radiation Survey and Site Investigation Manual (MARSSIM). This formula, which we dub the N13.30 rule, does not adequately account for the discrete nature of the Poisson distribution for paired blank (equal count times for background and sample) measurements, especially at low numbers of counts. We calculate the actual false positive rates that occur using the N13.30 DL formula as a function of a priori false positive rate a and background Poisson mean mu = rhot, where rho is the underlying Poisson rate and t is the counting time. False positive rates exceed a by significant amounts for alpha < or = 0.2 and mu < 100 counts, peaking at 25% at mu approximately equal to 0.71, nearly independent of alpha. Monte Carlo simulations verified calculations. Currie's derivation of the N13.30 DL was based on knowing a good estimate of the mean and standard deviation of background, a case that does not hold for paired blanks and low background rates. We propose one new decision rule (simply add 1 to the number of background counts), and we present six additional decision rules from various sources. We evaluate the actual false positive rate for all eight decision rules as a function of a priori false positive rate and background mean. All of the seven alternative rules perform better than the N13.30 rule. Each has advantages and drawbacks. Given these results, we believe that many regulations, national standards, guidance documents, and texts should be corrected or modified to use a better decision rule. PMID- 11414622 TI - A dose reconstruction of 60Co-contaminated window frames in a Taiwanese school. AB - Since 1992, hundreds of buildings in Taiwan were discovered to have 60Co contamination in the structural rebar. The contamination resulted from improper handling of 60Co-contaminated scrap metal in 1982 and 1983, which subsequently was recycled and used throughout Taiwan. Hsin-hsin Kindergarten school enrolled about 600 students over the 10-y period before the contamination was discovered. Hsin-hsin Kindergarten had three 60Co-contaminated steel window frames with measured dose rates on contact up to 150 microSv h(-1). In this study, a range of potential doses received by the Hsin-hsin Kindergarten students were estimated using ISOSHLD dose modeling software. ISOSHLD is a rapid, inexpensive screening tool to reconstruct dose ranges. To assess the potential risks to habitants of the school for the first year after construction, calculated dose rate ranges of 0.08 microSv h(-1) to 75.38 microSv h(-1) were then applied to the International Commission [corrected] on Radiation Protection 60 nominal detriment coefficients for stochastic effects. Risk estimates ranged from 1.46 x 10(-4) to 7.42 x 10(-4) excess fatal cancers per lifetime. PMID- 11414623 TI - Scab diameters on pig skin resulting from hot particle exposures under varying conditions. AB - The National Council on Radiation Protection and Measurements (NCRP) in NCRP Report Number 106 recommended a limit of 10(10) beta particles emitted from radioactive particles with sizes less than 1 mm (hot particles) to prevent acute deep ulceration. This recommendation was determined, in part, by regressing the diameter of the scabs induced by fissioned 235UC2 hot particles as a function of the logarithm of the number of beta particles emitted from the sources for one study. To validate this recommendation and the approach used by the NCRP, external irradiations of pig skin using radioactive sources of less than 600 microm in the largest dimension were carried out. The hot particles used included fissioned 235UC2 and activated 170Tm, 175Yb, and 46Sc. Results indicated a strong correlation between scab diameter and dose for scabs induced using fissioned 235UC2, activated 170Tm, and 46Sc, but not for 175Yb. The correlation value decreased with decreasing beta particle energy, with the exception of 46Sc, which had scabs with diameters greater than twice the maximum beta particle range. The larger scabs from 46Sc are thought to be due to dose contributions from the gamma rays. The results also give an ulceration threshold less than that given by NCRP to prevent acute deep ulceration. It was concluded that regression analysis of scab diameter as a function of either number of beta particles emitted from the hot particles or dose did not yield either precise or accurate thresholds but was useful in determining probable ranges of doses which lead to scab induction. PMID- 11414624 TI - Scab incidence on pig skin resulting from hot particle exposures under varying conditions. AB - External irradiations of pig skin using radioactive sources of less than 600 microm in the largest dimension (hot particles) were carried out. The objective of the study was to determine a threshold for scab induction. Hot particles used included fissioned 235U and activated 170Tm, 17SYb, and 46Sc with maximum beta particle energies of approximately 1.8 (average), 0.97, 0.47, and 0.35 MeV, respectively. The photon emissions from the fissioned 235U were about 1 MeV per disintegration. The photon emissions from 46Sc were 0.89 and 1.12 MeV, with 100% abundance. Photon emissions from 170Tm and 175Yb were negligible. Responses followed cumulative normal probability distributions; thus, no true thresholds could be determined. Hence, 10% and 50% scab incidence rates (ED10 and ED50, respectively) were determined using probit analysis. For dose averaged over 1 cm2 at a depth of 70 microm, the ED10 (and 95% confidence limits) were 5.1 (2.7-7.2) Gy for 46Sc, 1.3 (0.89-1.8) Gy for 175Yb, 2.8 (1.9-3.5) Gy for 170Tm, 8.5 (5.7 9.8) Gy for on-skin fissioned UC2, and 4.5 (0.9-7.2) Gy for off-skin fissioned UC2. The ED50 values were 12 (8.8-17) Gy for 46Sc, 6.0 (4.3-9.7) Gy for 175Yb, 5.9 (5.1-6.7) Gy for 170Tm, 11 (9.9-19) Gy for on-skin fissioned 235UC2, and 11 (6.2-14) Gy for off-skin fissioned 235UC2. PMID- 11414625 TI - Investigation of the air pressure characteristics influencing the variability of radon gas and radon progeny in domestic vernacular buildings. AB - Analysis of time-series data sets collected in vernacular buildings linked with radium source bedrock has identified a number of internal and external pressure characteristics linking meteorological parameters with the variability of radon gas and its progeny. The buildings' cellars built into the bedrock associated with the radium source have relatively high levels of radon concentration. These cellars have essentially stable microclimatic conditions, unlike the ground and upper levels of the buildings. Comparative radon concentration data collected from various comparable buildings suggest the need to distinguish between short and longer-term influences on radon concentrations. Water vapor pressure is inferred to be a principal determinant of the short-term variability of radon gas concentrations. Barometric pressure is suggested as determining the trend or general longer-term level of radon. Both of these pressure components are related to temperature. Wind speed appears to have a dual influence on radon variability: directly, through wind pressure relative to the ground and building structure particularly associated with low-pressure weather regimes; and indirectly, through changes to the water vapor pressure component. PMID- 11414626 TI - Reconstruction of doses from radionuclide inhalation for nuclear power plant workers using air concentration measurements and associated uncertainties. AB - This paper presents a case study to illustrate the influence of parameter uncertainties on calculating an internal radiation dose of one actual nuclear plant worker, alias Mr. X, as well as the utility of air sampling for internal dose reconstruction. Input probability distributions for air concentrations of radionuclides were derived from empirical air measurements taken by fixed area air samplers. The total internal dose was calculated by multiplying radionuclide intake by dose conversion factors in Monte-Carlo simulations. There is significant variability in dose conversion factors and uncertainty in the estimated concentrations of radionuclides in the air to which Mr. X was exposed. The high variability and uncertainty of the model parameters contributed to large ranges of predicted internal doses for Mr. X. Two-dimensional Monte-Carlo simulations were conducted to separate contributions of the uncertainty and variability. Sensitivity analysis was conducted to determine which of the input parameters contributed most to uncertainty in internal dose estimates. Our analysis suggests that the uncertainty resulting from use of general air surveys contributes more to the internal dose ranges than the variability from DCFs and other population-derived parameters. Reduction of the uncertainty in reconstructed internal dose can be achieved by using personal air sampling and/or individual bioassays and regular whole-body counting. PMID- 11414627 TI - Determination of 232Th in urine by ICP-MS for individual monitoring purposes. AB - Thorium is naturally occurring in various ores used for industrial purposes and has numerous applications. This paper sets out to investigate urine analysis as a suitable monitoring approach for workers potentially exposed to thorium. Due to its biokinetic behavior and its low solubility, urinary concentrations are generally very low, requiring therefore high sensitivity analytical methods. An analytical procedure has been developed for detecting 232Th concentrations of below 1 mBq L(-1) quickly and easily. Due to the long half-life (1.41 x 10(10) y) of 232Th, the potential of a procedure based on urine sample dilution and ICP-MS (inductively coupled plasma-mass spectrometry) measurement was investigated first. Two dilution factors were chosen: 100, which is more suitable for long term measurement trials, and 20, which increases sensitivity. It has been shown that a 100-fold dilution can be used to measure concentrations of below 1 mBq L( 1), whereas a 20-fold one can be used to reach concentrations of below 0.06 mBq L(-1). Then, on the basis of the limitation of the procedure based on urine dilution, the suitable field of application for the different procedures (100 fold and 20-fold dilution and also a chemical purification followed by an ICP-MS measurement) was determined in relation to monitoring objectives. PMID- 11414628 TI - Retinal damage induced by red diode laser. AB - Widespread use of compact, low-cost diode lasers (pointers and illuminators) has ushered in an era where large numbers of the general public are accidentally or deliberately exposed to low-power laser radiation. The objectives of this study are both to determine the primate retinal lesion threshold for exposure to 650 nm diode laser radiation and to examine the risks of retinal damage from low-level (sub-threshold) ocular exposures. To this end, the ED50 and ED10 damage thresholds, their fiducial limits, and the slopes of the probability vs. dose curves have been examined in detail. In addition to conventional fundoscopy, exposed eyes were examined by confocal scanning laser ophthalmoscopy, fluorescein angiography, and histopathology at both the light and electron microscopic levels in attempts to discern tissue disruption following exposures below the ophthalmoscopic ED50 threshold dose. These alternative observation techniques did not identify detectable tissue disruption following exposures below the ophthalmoscopic lesion threshold dose. PMID- 11414629 TI - Rapid D-dimer testing and pre-test clinical probability in the exclusion of deep venous thrombosis in symptomatic outpatients. AB - We assessed the performance of three rapid D-dimer tests (Auto Dimertest, VIDAS and Tinaquant) in combination with a pretest clinical probability model for deep venous thrombosis (DVT) in 106 consecutive outpatients with suspected DVT. Contrast venography or colour-coded duplex ultrasonography demonstrated the presence of DVT in 47 patients (14 distal DVT and 33 proximal DVT). First, we assessed the accuracy indices for different cut-off levels of the rapid D-dimer tests. Sensitivity was found to be 97.9-100%, negative predictive value (NPV) was 96.3-100%, and the exclusion rate was 24.5-31.1%. Next, the patients were grouped according to the pre-test clinical probability model in categories with low, moderate or high probability. In patients with a low pre-test probability, DVT would have been directly ruled out and the patients would not have undergone further investigations. In patients with a moderate probability, D-dimer testing and, in the case of a positive result, objective testing would have been performed and, in the case of a negative result, they would have been ruled out of having DVT. Patients with high probability would directly have undergone objective tests for DVT. The combination with the pre-test clinical probability model improved the exclusion rate (43.5-44.6%), whereas sensitivity (97.5-100%) and NPV (97.6-100%) remained roughly unchanged. The combination of rapid D-dimer tests with a pre-test clinical probability model may help to reduce unnecessary work-up in patients with suspected DVT. PMID- 11414630 TI - Hemoglobin enhances tissue factor expression on human malignant cells. AB - Tissue Factor (TF) is a transmembrane glycoprotein that complexes with factor VII/activated factor VII to initiate blood coagulation. TF may be expressed on the surface of various cells including monocytes and endothelial cells. Over expression of TF in human tumor cell lines promotes metastasis. We recently showed that hemoglobin (Hb) forms a specific complex with TF purified from human malignant melanoma cells and enhances its procoagulant activity (PCA). To further study this interaction, we examined the effect of Hb on the expression of TF on human malignant (TF+) cells and KG1 myeloid leukemia (TF-) cells. Human melanoma A375 and J82 bladder carcinoma cells, which express TF at moderate and relatively high levels, respectively, were incubated with varying concentrations (0-1.5 mg/ml) of Hb. After washing, cells were analyzed for Hb binding and TF expression using flow cytometry and confocal microscopy. Hb bound to the cells in a concentration-dependent manner, and increased both TF expression and PCA. The human A375 malignant melanoma cells incubated with Hb (1 mg/ml) expressed up to six times more TF antigen than cells without Hb. This increase in TF expression and PCA of intact cells incubated with Hb was significantly inhibited by cycloheximide at a concentration of 10 microg/ml (P < 0.01). An increase in total cellular TF antigen content was demonstrated by specific immunoassay. In contrast, Hb (5 mg/ml) did not induce TF expression and PCA on KG1 cells as determined by flow cytometry and TF (FXAA) activity. We conclude that Hb specifically binds to TF-bearing malignant cells and increases their PCA. This effect seems to be at least partly due to de novo synthesis of TF and increased surface expression. However, the exact mechanism by which Hb binds and upregulates TF expression remains to be determined. PMID- 11414631 TI - The impact of oral anticoagulant therapy, factor VIII level and quality of factor V-deficient plasma on three commercial methods for activated protein C resistance. AB - Several methods are now available for the laboratory assessment of activated protein C resistance (APCR). In this study, we evaluated two activated partial thromboplastin time-based assays [Coatest activated protein C (APC) and Diagen protein C activator (PCA)], with and without predilution of test plasma in factor V-deficient plasma (FVdp) and an amidolytic assay (Immuno Ltd, Vienna, Austria). Testing plasmas from normal volunteers who had received 1-deamino-8-D-arginine vasopressin (DDAVP) also assessed the effect of elevated factor VIII on APCR. In the unmodified clotting tests, the Coatest kit gave overlapping results for normal and heterozygous FV:Q506 samples; some FV:Q506 samples on oral anticoagulant therapy (OAT) were misclassified as normal, and some normal samples with high factor VIII levels would be classified as APC resistant. The unmodified Diagen kit correctly classified these three types of sample, but had the disadvantage that prolonged PCA clotting times gave serious problems with instrument end-point detection. Both kits modified by diluting the samples in FVdp correctly classified all the samples, as well as samples from patients with lupus anticoagulant (LA) and patients receiving heparin. The Immunochrom kit correctly classified the normal and FV:Q506 samples, but would have misclassified most normal persons on OAT as well as some patients with LA or receiving heparin therapy as APC resistant. PMID- 11414632 TI - Plasma levels of factor XIIa and factor VIIa are increased but not related in primary hyperlipidaemia. AB - The lipolysis of triglyceride-rich lipoproteins may provide a surface that supports the activation of factor XII (FXII) with subsequent activation of factor VII (FVII). Plasma levels of activated FVII (FVIIa) but not activated FXII (FXIIa) are increased in the post-prandial state when there is a transient increase in triglyceride levels. We compared plasma levels of FXIIa antigen in control subjects (n = 33) and in patients with chronically elevated lipids (primary hyperlipidaemia, n = 49), with FVIIa and markers of thrombin generation. Results are given as median (first and third quartiles). Plasma levels of FXIIa [2.34 (1.68-3.32) ng/ml versus 1.53 (0.93-1.86) ng/ml, P = 0.0002], FVIIa [3.02 (2.15-4.64) ng/ml versus 2.20 (1.66-2.56) ng/ml, P = 0.0004], thrombin antithrombin complexes [3.08 (2.16-5.54) microg/I versus 2.13 (1.46-2.84) microg/l, P = 0.005] and prothrombin fragment 1 + 2 (Pro F1 + 2) [1.28 (1.08 1.50) nmol/l versus 0.92 (0.65-1.08) nmol/l, P = 0.0001] were increased compared with controls irrespective of the type of hyperlipidaemia. In hyperlipdaemic subjects, levels of Pro F1 + 2 were correlated with FVIIa (r = 0.56, P = 0.0002) and FXIIa (r = 0.31, P = 0.03). These results suggest increased activation of both FVII and FXII in hyperlipidaemic subjects, which correlates with increased thrombin generation. Given the lack of correlation between levels of FXIIa and FVIIa, it remains to be established whether the increase in FXIIa is responsible for increased FVIIa activity in this subject group. PMID- 11414633 TI - Expression of factor VII in the liver of patients with liver disease: correlations with the disease severity and impairment in the hemostasis. AB - Factor VII (FVII) plasma levels in patients with liver disease may be below the normal range. However, no data are available on FVII expression in liver biopsies from patients with liver diseases other than cirrhosis. We have analyzed the expression of FVII by in situ hybridization in liver biopsies from 50 patients in comparison with the procoagulant activity of FVII, and with the plasma levels as activated FVII (FVIIa) and FVII antigen. The level of FVIIa was significantly lower in stage 4 liver fibrosis patients than in the remaining ones (P < 0.05). The percentage of hepatocytes expressing FVII was significantly lower in stage 4 liver fibrosis patients (4.1+/-1.3%) than in stage 3 (22.7+/-6.1%), stage 2 (31.5+/-6.1%), stage 1 (43.7+/-8.2%) and stage 0 patients (63.8+/-4.4%) (P < 0.001). These percentages correlated inversely in a statistically significant way with the histological activity index and the liver function tests. We have demonstrated that the FVIIa plasma levels in patients with chronic liver disease other than cirrhosis may be below the normal range in the absence of blood coagulation impairment. The percentage of hepatocytes expressing FVII decreases as the severity of liver damage increases. PMID- 11414634 TI - The factor V HR2 haplotype: prevalence and association of the A4070G and A6755G polymorphisms. AB - Recently, a polymorphism was identified in exon 25 of the factor V gene that is possibly a functional candidate for the HR2 haplotype. This haplotype is characterized by a single base substitution named R2 (A4070G) in the B domain of the protein. A mutation (A6755G; 2194Asp-->Gly) located near the C terminus has been hypothesized to influence protein folding and glycosylation, and might be responsible for the shift in factor V isoform (FV1 / FV2) ratio. This study investigated the prevalence of these two factor V HR2 haplotype polymorphisms in a cohort of normal blood donors, patients with osteoarthritis and women with complications during pregnancy, and in families of factor V Leiden individuals. A high allele frequency for the two polymorphisms was found in the blood donor group (6.2% R2, 5.6% A6755G). No significant difference in allele frequency was observed in the clinical groups (obstetric complications and osteoarthritis, 4.1 4.9% for the two polymorphisms) when compared with that of healthy blood donors. We confirm that the factor V A6755G polymorphism shows strong linkage to the R2 allele, although it is not exclusively inherited with the exon 13 A4070G variant and can occur independently. PMID- 11414635 TI - Recurrent haemoperitoneum in a mild von Willebrand's disease combined with a storage pool deficit. AB - Haemoperitoneum secondary to haemorrhagic corpus luteum has been described in severe bleeding disorders such as afibrinogenaemia, type 3 von Willebrand's disease and patients under oral anticoagulation. We have studied one patient who presented three episodes of severe bleeding at ovulation, requiring surgery twice, with the diagnosis of mild von Willebrand's disease and mild storage pool deficiency. Mild von Willebrand's disease (associated with other thrombopathies or coagulopathies) should be considered in this pathology, although physicians would prefer to find a severe haemorrhagic disorder as the underlying condition in these cases. PMID- 11414636 TI - Recurrent severe bleeding from gastrointestinal angiodysplasia in a patient with von Willebrand's disease, controlled with recombinant factor VIIa. AB - A patient with von Willebrand's disease had recurrent gastrointestinal bleeding from angiodysplasia, with inadequate response to von Willebrand factor substitution, medical and endoscopic treatment, and resection of affected bowel. Frequent blood transfusions were required. She started home treatment with recombinant activated factor VII (rFVIIa) at the onset of bleeding, in addition to her standard therapy. From then on, bleeds could be controlled rapidly and no more blood transfusions were needed. We conclude that rFVIIa is effective in this case of angiodysplasia and might be a therapeutic option in similar patients. PMID- 11414637 TI - Successful use of recombinant factor VIIa in a haemophiliac with inhibitor undergoing cataract surgery. AB - A 40-year-old patient with severe haemophilia A and an inhibitor against factor VIII underwent a cataract extraction under local anaesthesia. Recombinant activated factor VII was use to achieve haemostasis. The procedure was successful. Neither bleeding complications nor side effects occurred. PMID- 11414638 TI - Performances of a new, automated latex assay for the exclusion of venous thromboembolism. AB - The performance of a new latex-enhanced turbidimetric assay, D-Dimer PLUS, has been evaluated with two analyzers performing various coagulation assays: the BCS Analyzer and the BCT Analyzer. A precision study showed total coefficients of variation ranging from 2.7 to 11.1% with the BCS Analyzer and from 2.5 to 6.6% with the BCT Analyzer. We investigated the ability of D-Dimer PLUS to exclude venous thromboembolism in 312 outpatients suspected of either pulmonary embolism or deep venous thrombosis. Three months follow-up was available for all patients. With the BCS Analyzer, we determined a cut-off value of 190 ng/ml, which gave a sensitivity of 97.9% [95% confidence interval (CI), 92.6-99.7%], a specificity of 37.9% (95% CI, 30.9-43.8%) and a negative predictive value of 97.6% (95% CI, 91.7 99.7%). With the BCT Analyzer, at a cut-off value of 130 ng/ml, sensitivity was 96.8% (95% CI, 91.0-99.3%), specificity was 45.2% (95% CI, 38.5-51.2%) and the negative predictive value was 97% (95% CI, 91.6-99.4). This new assay is fast and fully automated, and its performance is suitable to exclude venous thromboembolism. Management studies should be performed to assess its utility. PMID- 11414639 TI - Effect of lupus anticoagulants on INR with different thromboplastins. PMID- 11414640 TI - Late detection of neuroblastoma in a patient with prolonged cerebellar ataxia without opsoclonus. AB - A 19-month-old boy presented with acute-onset cerebellar ataxia following immunisation. Ataxia was prolonged with a fluctuating course over a period of two years. Opsoclonus was never observed. Extensive diagnostic studies were negative during this time. Finally, a neuroblastoma was discovered. Ataxia disappeared completely during polychemotherapy. This report suggests that occult neuroblastoma must be considered in any child with unexplained prolonged cerebellar ataxia. Specific investigations are recommended for such cases. PMID- 11414641 TI - Mitochondrial respiratory chain deficiency revealed by hypothermia. AB - We observed a 17-month-old girl with profound and initially isolated episodes of hypothermia. Thereafter, she developed growth delay, repetitive corneal and bone lesions. Persistent hyperlactataemia in plasma and in CSF prompted us to investigate respiratory chain enzymes. A deficit in respiratory chain complexes III and IV was demonstrated in isolated skeletal muscle mitochondria, circulating lymphocytes and fibroblasts by spectrophotometric and polarographic studies. Moreover, UCP3 mRNA expression in muscle was decreased. PMID- 11414642 TI - Myopathy in a patient with chromosome 22q11 deletion. PMID- 11414643 TI - Characteristic clinical features of idiopathic neuralgic amyotrophy in childhood. PMID- 11414644 TI - Oppenheim's myatonia congenita. AB - In 1900 the famous German neurologist Hermann Oppenheim published a two-page article on "Myatonia congenita" which led to a long-lasting and confusing discussion in neurology and neuropediatrics, and which was extensively quoted throughout the 20th century. As this article is only available in German, an English translation is presented here. Further publications of Oppenheim on the same subject are mentioned and some comments are made on the impact of his seminal article. It had the merit of drawing the attention to the existence of congenital muscle diseases in children but its impact would be better understood by the influence Oppenheim had on his time, as he was one of the leading neurologists at the end of the 19th and beginning of the 20th century. PMID- 11414645 TI - Near-total absence of the cerebellum. AB - We report five cases of near-total absence of the cerebellum with accompanying pontine hypoplasia. The cerebellar remnant in each case comprised only antero superior masses, the posterior fossa being otherwise fluid filled. Three of these patients, two teenagers and an infant, presented a fairly consistent clinical and neuroradiological phenotype, and a few similar cases are recorded in the literature. The cerebellar remnant was irregular and asymmetrical, and no ventral pontine prominence was discernible. In at least the older two, cerebellar motor functions were not greatly compromised, and intellectual handicap was of a mild degree. We propose that these cases represent a distinct entity of "near-total absence of the cerebellum with flat ventral pons, and relatively mild clinical affection". All cases have been sporadic, implying that the risk of recurrence within a family may be low. Quite different clinical pictures, of considerably greater severity, are demonstrated in the remaining two cases. One had pontocerebellar hypoplasia type 2, while the other had a complex cerebellar and cerebral malformation. PMID- 11414646 TI - Brain development (sulci and gyri) as assessed by early postnatal MR imaging in preterm and term newborn infants. AB - OBJECTIVE: To investigate fetal brain development in vivo using early postnatal cranial MRI in term and preterm newborn infants. STUDY DESIGN: 51 infants, 1.5-T whole-body system, extremity coil, spin-echo images obtained in all three planes (T1- and T2-weighted). Independent review by two neuroradiologists (blinded for gestational age and medical history) regarding the development of 12 sulci and 10 gyri using a modified scoring system. Gestational age was obtained either by ultrasound or from the first day of the last menstrual period. EXCLUSION CRITERIA: head circumference <10th or >90th percentile, cerebral malformation, chromosomal/metabolic disorder. RESULTS AND CONCLUSION: Age at study ranged from 23 5/7-43 0/7 postmenstrual weeks. Brain maturation starts in the central area and proceeds towards the parieto-occipital cortex. The frontal cortex develops last. Transition to degrees 1, degrees 2 and, degrees 3 starts at week 25, 32 and 35 in the central cortex and is completed at week 34, 36 and 39, respectively. Our data compare favourably with the two previously published reports about brain maturation. Early MR imaging seems therefore to be suited to study maturation of the fetal brain. This may be of pathophysiological relevance in neonatal intensive care and neurological follow-up studies. PMID- 11414647 TI - High microvessel density in primitive neuroectodermal brain tumors of childhood. AB - Microvessel density (MVD), a measure of tumor angiogenesis, has been shown to correlate significantly with overall and progression-free survival outcomes in various cancers including astrocytic brain tumors. To assess if the MVD is an independent prognostic factor in primitive neuroectodermal tumors (PNET) of the central nervous system, formalin-fixed paraffin-embedded tumor sections of 78 children with PNET were studied by CD34 immunohistochemistry to highlight endothelial cells. Microvessel density was determined in the most active area of neovascularization according to well-established methods. While it was shown that MVD showed considerable inter-tumor variability (median 75; range 20-345 microvessels per 0.7 mm2 field), no significant associations were found between MVD and metastasis or survival outcomes. We conclude that many PNETs are highly vascular CNS tumors, indicating potent angiogenic activity. Therefore, these tumors would be good candidates for antiangiogenic strategies. However, MVD determined in the most active area of neovascularization is not a predictor of metastatic potential or survival outcomes in childhood PNET. PMID- 11414648 TI - Parenchymal brain injury in the preterm infant: comparison of cranial ultrasound, MRI and neurodevelopmental outcome. AB - AIM: Magnetic resonance imaging (MRI) is increasingly being used in high-risk preterm neonates. Cranial ultrasound (US) was compared with MRI in preterm patients with parenchymal injury and related to neurodevelopmental outcome. PATIENTS AND METHODS: Studies were performed in 61 patients. Twelve infants with normal US (Group 1) had an MRI within the first 4 weeks of life (early MRI), and 10 also at term age (late MRI). Eight out of 20 infants with intraventricular haemorrhage with parenchymal involvement (IVH + PI) (Group 2) had an early as well as a late MRI and 12 a late MRI. Of the 20 patients with cystic periventricular leukomalacia (c-PVL) (Group 3), 7 had an early MRI, 1 had an MRI on both occasions and 12 had a late MRI. All 9 children with focal infarction (FI) (Group 4) had a late MRI. RESULTS: MRI was conform with cranial US in Group 1. Early MRI in Group 2 showed contralateral c-PVL in one infant and an additional contralateral occipital parenchymal haemorrhage and blood in the posterior fossa in another infant. Late MRI showed an asymmetrical posterior limb of the internal capsule (PLIC) (n=6), which predicted later hemiplegia. Early MRI in Group 3 showed more cysts (n = 5), punctate white matter lesions (n = 6), lesions in the basal ganglia (n = 1) and once involvement of the cerebellum. Late MRI showed involvement of the centrum semiovale (n = 2) lesions in the basal ganglia (n = 2) and bilateral abnormal signal intensity of the PLIC in 7 infants who all went on to develop cerebral palsy. In Group 4 MRI showed signal intensity changes suggestive of cystic lesions compared to persisting echogenicity on US (n = 3) and an asymmetrical PLIC (n = 5), which predicted hemiplegia in 4. CONCLUSION: Early MRI especially provided additional information in those with c PVL. MRI at term age could assess the PLIC, which was useful in children with unilateral parenchymal involvement, for prediction of subsequent hemiplegia and, to a lesser degree, in bilateral c-PVL for prediction of diplegia or quadriplegia. PMID- 11414649 TI - Permanent brain damage following acute clonidine poisoning in Munchausen by proxy. AB - A child presented with recurrent episodes of lethargia for which he underwent several hospital admissions and investigations. A further episode culminated in respiratory arrest and hypoxic ischemic encephalopathy with permanent mental regression. Eighteen months later the mother was discovered while providing clonidine pills to the child; the mother appears to feature a Munchausen syndrome by proxy. PMID- 11414650 TI - Scheie syndrome presenting as myopathy. AB - We investigated two brothers with Scheie syndrome whose only complaint was exercise intolerance. In the quadriceps muscle biopsy of both patients, between the normal muscle fibres an increased number of markedly swollen periodic acid Schiff-positive fibroblasts were seen. Ultrastructurally, these cells showed an accumulation of enlarged lysosomes, partly filled with electro-dense material. We hypothesize that the accumulation of pathological fibroblasts may interfere with intramuscular force transmission resulting in exercise intolerance. PMID- 11414651 TI - Proton MR spectroscopy reveals lactate in infantile neuroaxonal dystrophy (INAD). AB - Changes of cerebral metabolites detected by proton MR spectroscopy in two cases of infantile neuroaxonal dystrophy are described. A 6 11/12-year-old boy and a girl (aged 4 1/12 years at the first and 5 2/12 years at the second examination) with infantile neuroaxonal dystrophy were investigated by magnetic resonance imaging and spectroscopy of the basal ganglia. The signal intensity of the cerebellar cortex was increased on T2-weighted, proton density, and fluid attenuated inversion recovery images. The long echo time (135 ms) spectra revealed the presence of lactate in the basal ganglia of both cases in all investigations. The N-acetylaspartate/creatine ratio was reduced in Case 1 and in the second investigation of Case 2. The choline/creatine ratio was always increased. As the diagnosis of infantile neuroaxonal dystrophy is made by a synopsis of various clinical, neuropathological, neurophysiological, and neuroradiological data, the presence of lactate in the basal ganglia spectra may help to narrow down the diagnosis and can support the decision to perform more invasive diagnostic procedures (such as biopsies of skin, conjunctiva or even of the brain). PMID- 11414652 TI - Differential antagonism by naloxone benzoylhydrazone of the activation of inward rectifying K+ channels by nociceptin and a mu-opioid in rat periaqueductal grey slices. AB - A novel receptor, the opioid receptor-like orphan receptor (ORL1), is homologous to, but distinct from, classical opioid receptors. Although initially developed as an opioid receptor ligand, naloxone benzoylhydrazone (NalBzOH) is one of the few antagonists at ORL1. The present electrophysiological study of the effects of NalBzOH on the activation of ORL1 and mu-opioid receptors was performed in brain slices of the ventrolateral periaqueductal grey (PAG), a crucial site for opioid induced supraspinal analgesia. Both orphanin FQ/nociceptin (OFQ/N), an ORL1 agonist, and [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin (DAMGO), a mu-opioid receptor agonist, activated inwardly rectifying K+ (Kir) channels in the ventrolateral PAG. Of the neurons tested, 96% responded to OFQ/N, but only 65% to DAMGO. NalBzOH (3-30 microM) antagonized the effect of OFQ/N competitively with a pA2 of 5.67. NalBzOH also antagonized, but more potently and non-competitively, the effect of DAMGO. In contrast, NalBzOH did not affect baclofen-induced activation of Kir channels. NalBzOH alone, at concentrations up to 30 microM, had little effect on this inwardly rectifying channel. It is concluded that NalBzOH antagonizes the activation of Kir channels mediated by both ORL1 and mu-opioid receptors in the ventrolateral PAG. It acts not only as a competitive antagonist at ORL1, but also as a more potent and non-competitive antagonist at mu-opioid receptors. PMID- 11414653 TI - Chromanol 293B, a blocker of the slow delayed rectifier K+ current (IKs), inhibits the CFTR Cl- current. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) and the sulphonylurea receptor subunit (SUR) of the KATP channel are both members of the ATP-binding cassette (ABC) protein superfamily. Many compounds that open or block the KATP channel by binding to SUR also inhibit the CFTR Cl- current (ICFTR); an example in point is the chromanol-type KATP channel opener, cromakalim. The structurally related chromanol 293B (trans-6-cyano-4-(N-ethylsulfonyl-N methylamino)-3-hydroxy-2,2-dimethyl-chromane), a blocker of the slow component of the delayed rectifier K+ current (IKs) in the heart, is also a weak inhibitor of KATP. This suggests that 293B may affect also ICFTR- We have addressed this question with human CFTR expressed in Xenopus oocytes. In two-electrode voltage clamp experiments, 293B inhibited ICFTR with an IC50-value of 19 microM and Hill coefficient of 1.0; the inhibition was weakened by increasing concentrations of isobutyl-methylxanthine (IBMX). Patch-clamp recordings gave an IC50-value of 30 microM but showed a unusual variability in the sensitivity to 293B. The data show that 293B inhibits ICFTR and suggest that the mechanism of inhibition may depend on the phosphorylation state of the CFTR protein. The concentrations required for inhibition of ICFTR are three- to fivefold higher than those reported for inhibition of KvLQT1 + minK expressed in Xenopus oocytes. Since CFTR is expressed also in cardiac myocytes, the effects of 293B in these cells must be analysed with caution. PMID- 11414654 TI - Inhibitory effect of SM-20550, an Na+/H+ exchange inhibitor, on accumulation of leukocytes in ischemia and reperfusion. AB - The effect of an Na+/H+ exchange inhibitor, SM-20550, on the adhesion, emigration and accumulation of leukocytes was studied in ischemia and reperfusion injury models using rat mesenteric venules or rabbit heart. Anesthetized rats underwent occlusion of the superior mesenteric artery (20 min) followed by reperfusion (60 min). After ischemia and reperfusion, the numbers of adherent and emigrated leukocytes increased significantly. Bolus intravenous administration of SM-20550 reduced the numbers of adherent and emigrated leukocytes in a dose-dependent manner, with statistical significance at dosages >0.1 mg/kg. Anesthetized rabbits underwent occlusion of the coronary artery (30 min) followed by reperfusion (5 h). Intravenous administration of SM-20550 before ischemia significantly reduced the neutrophil accumulation in the area-at-risk by 46% and reduced the infarct size by 38%. These results suggest that the inhibitory effect of SM-20550 on the neutrophil accumulation could be one of the mechanisms by which this drug limits the severity of infarctions. PMID- 11414655 TI - Effects of the antiarrhythmic agent EGIS-7229 (S 21407) on calcium and potassium currents in canine ventricular cardiomyocytes. AB - Based on earlier pharmacological studies performed using conventional microelectrodes EGIS-7229 (S 21407), the novel antiarrhythmic candidate, was suggested to have a combined mode of action in cardiac tissues isolated from various mammalian species. In order to characterize the electrophysiological effects of the compound, its effects on calcium and potassium currents of isolated canine ventricular cardiomyocytes were studied in the present work using the whole cell configuration of the patch clamp technique. L-type Ca current (ICa) was significantly depressed by EGIS-7229 at concentrations of 3 microM or higher with no concomitant changes in the voltage-dependence of activation and time course of inactivation of ICa. The drug reversibly suppressed the rapid component of the delayed rectifier K current (IKr) in a concentration-dependent manner, having a K0.5 value of 1.1+/-0.1 microM and a slope factor of close to unity (1.23+/-0.16), indicating that probably one single binding site of high affinity may be involved in binding of EGIS-7229 to the IKr channel. In contrast, no changes in the slow component of the delayed rectifier K current (IKs) was observed with the compound up to the concentration of 100 microM, even if the current was fully activated by 8-bromo-cAMP. At a concentration of 10 microM or higher, EGIS-7229 caused also a moderate but significant reduction in the inward rectifier K current (IK1) and the transient outward K current (Ito) with no change in the voltage-dependence of activation and steady-state inactivation of Ito. Present results indicate that EGIS-7229 can be considered as a selective IKr blocker at low (1 microM) concentration; however, its combined (class III + IV) mechanism of action is evident at concentrations of 3 microM or higher. Suppression of ICa may explain the lack of development of early afterdepolarizations in the presence of EGIS-7229, predicting a relatively safe clinical application in contrast to pure class III compounds. PMID- 11414656 TI - Role of cyclic nucleotide phosphodiesterase isoenzymes in contractile responses of denuded rat aorta related to various Ca2+ sources. AB - We have examined the cyclic nucleotide phosphodiesterase isoforms (PDE) involved in the contractile response of rat aorta to different agonists and different experimental procedures for use in functional studies. The inhibitory effect of AAL 05 on the different PDEs isolated from bovine aortic smooth muscle was examined. Compound AAL 05 appeared to be a selective PDE3 inhibitor. We analyzed the ability of the non-selective inhibitor IBMX (3-isobutyl-1-methylxanthine) and the isoenzyme selective inhibitors nimodipine (type 1), AAL 05 (6-(N-methyl-N cyclohexyl butyl carboxamide) quinolin-2-one) and SK&F 94120 (5-(4 acetamidophenyl) pyrazin-2(1H)-one; type3), rolipram (type4) and zaprinast (type5) to affect the contractile responses of denuded rat aortic rings to KCl (80 mM) and noradrenaline (NA, 1 microM) in the presence or absence of extracellular Ca2+. Rolipram (10-100 microM) and zaprinast (1-100 microM) failed to relax the aortic strips, but IBMX (0.1-30 microM), nimodipine (1 fM10 microM), AAL 05 (0.01-100 microM) and SK&F 94120 (0.1-100 microM) produced a concentration dependent relaxation or inhibition of contractile responses to the different agonists, but the pIC50 obtained for each inhibitor was different depending on the experimental procedure. Except for nimodipine (a Ca2+ channel blocker), all the PDE inhibitors showed the following rank of potency: pIC50 on NA-induced contractions in Ca2+-free medium > pIC50 on NA-induced contractions in Ca2+ containing solution > pIC50 on depolarizing solution-induced contraction. This ranking apparently depends on the differences in the Ca2+ sources. We obtained a good correlation between the pKi of PDE3 inhibitors in biochemical studies and the pIC50 on NA-induced contraction in Ca2+-free medium. In conclusion, PDE1 and PDE3 isoenzymes play an important role as modulators of rat aortic smooth muscle contractility regardless of the experimental procedure used. Since intracellular mechanisms are more dependent on PDE activity, experimental procedures performed in absence of extracellular calcium are the most suitable for analyzing the modulatory role of PDE inhibitors. PMID- 11414657 TI - The cloned human 5-HT7 receptor splice variants: a comparative characterization of their pharmacology, function and distribution. AB - Serotonin (5-hydroxytryptamine, 5-HT) receptor pre-mRNA is alternatively spliced in human tissue to produce three splice variants, h5-HT7(a), h5-HT7(b) and h5 HT7(d), which differ only in their carboxyl terminal tails. Using membranes from transiently and stably transfected HEK293 cells expressing the three recombinant h5-HT7 splice variants we compared their pharmacological profiles and ability to activate adenylyl cyclase. Using PCR on cDNA derived from various human tissues, the 5-HT7(a) and 5-HT7(b) splice variants were detected in every tissue examined. The h5-HT7(d) splice variant was detected in 13 of 16 tissues examined, with predominant expression in the heart, small intestine, colon, ovary and testis. All three h5-HT7 splice variants displayed high affinity binding for [3H]5-HT (pKd=8.8-8.9) in the presence and absence of 100 microM GTP and had similar binding affinities for all 17 ligands evaluated. In HEK293 cells expressing similar, high levels of receptor (approximately 10,000 fmol/mg protein), 5-CT (5 carboxamidotryptamine), 5-MeOT (5-methoxytryptamine) and 5-HT were full agonists while 8-OH-DPAT ((2R)-(+)-8-hydroxy-2-(di-n-propylamino)tetralin) was a partial agonist with relative efficacy of approximately 0.8. Even at this high receptor level, EC50 values for stimulation of adenylyl cyclase were 10- to 50-fold higher than the Kd values, indicating a lack of spare receptors. No significant differences in coupling to adenylyl cyclase were observed between the three splice variants over a wide range of receptor expression levels. For antagonists, binding affinities determined by displacement of [3H]5-HT binding and by competitive inhibition of 5-HT-stimulated adenylyl cyclase activity were essentially identical amongst the splice variants. These studies indicate that the three human splice variants are pharmacologically indistinguishable and that modifications of the carboxyl tail do not influence coupling to adenylyl cyclase. PMID- 11414658 TI - Activation of store-operated Ca2+ entry in RBL cells without the contribution of protein kinases. AB - The role of protein kinases on store-operated Ca2+ entry in rat basophilic leukaemia cells (RBL) has been studied using the whole-cell configuration of the patch-clamp technique and Ca2+ imaging with fura-2. Specific inhibitors of tyrosine kinase (lavendustin A), mitogen-activated protein (MAP) kinase (SB 203580, PD 98059), Ca2+/calmodulin-dependent kinase (CaMK, KN-62, KN-93) and protein kinase C (PKC, bisindolylmaleimide I) had no significant effect on peak current amplitude and time constant of activation. Likewise, the broad spectrum kinase blockers H-7 and staurosporine did not alter Ca2+ entry compared to control recordings. Store-mediated Ca2+ entry was unaffected if intracellular ATP was substituted by either adenosine 5'-O-(2-thiodiphosphate) (ADPbetaS) or adenylyl-imidodiphosphate (AMP-PNP). Similarly, buffering intracellular Mg2+, an essential cofactor for protein kinases, had no effect on Ca2+ influx. These results indicate that protein phosphorylation by various kinases is not required for the activation of the store-operated Ca2+ current in RBL cells. PMID- 11414659 TI - Species- and agonist-dependent differences in the deactivation-kinetics of P2X7 receptors. AB - In this study we have expressed recombinant P2X7 receptors in HEK293 cells and examined the reasons for the species- and agonist-dependent differences in the time taken for the closure of the P2X7 receptor ion-channels after agonist removal. Channel closure times, measured in electrophysiological studies or by measuring cellular permeability to ethidium cations, were slower at rat than at human or mouse P2X7 channels following washout of the P2X7 agonist 2'- and 3'-O (4-benzoylbenzoyl)-ATP (BzATP). In contrast, there were no species differences in channel closure times when ATP was the agonist. BzATP was more potent than ATP at the three species homologues and exhibited highest potency for rat P2X7 receptors suggesting that channel closure time was related to agonist potency. Furthermore, BzATP potency for the P2X7 receptor could be modified by changing extracellular ionic concentrations or by mutating the receptor and modifications which increased agonist potency also increased the time taken for channel closure. The dependence of channel closure time on agonist potency suggests it reflects agonist dissociation from the P2X7 receptor rather being an intrinsic property of the ion-channel. Consistent with this, our previous studies have shown that agonist potency increases after repeated agonist applications and in this study channel closure time at rat P2X7 receptors increased after repeated agonist applications. Overall these results suggest that the species differences in channel closure times reflect differences in agonist dissociation rates which arise as a consequence of the marked species differences in agonist potency. PMID- 11414660 TI - In vitro and in vivo uroselectivity of B8805-033, an antagonist with high affinity at prostatic alpha1A- vs. alpha1B- and alpha1D-adrenoceptors. AB - We have investigated the pharmacological properties of B8805-033 [(+/-)- 1,3,5 trimethyl-6-[[3-[4-((2,3-dihydro-2-hydroxymethyl)-1,4-benzodioxin-5-yl)-1 piperazinyl]propyl] amino]-2,4(1H,3H)-pyrimidinedione], a new alpha1A adrenoceptor (AR) selective antagonist. In radioligand binding studies, B8805-033 was 150- to 1200-fold selective for alpha1A-ARs (pKi rat cerebral cortex 8.70, cloned human receptor 7.71) relative to alpha1B-ARs (pKi rat cerebral cortex 5.60, rat liver 5.39, cloned human receptor 5.16) and alpha1D-ARs (pKi cloned human receptor 5.49). B8805-033 inhibited noradrenaline (NA) induced contractions mediated by alpha1A-ARs in rat vas deferens and rabbit and human prostate (pA2 7.62-8.40) much more potently than those mediated by alpha1B-ARs in guinea pig and mouse spleen or by alpha1D-ARs in rat aorta and pulmonary artery (pA2 5.21 5.52). With the exception of a high agonist affinity at 5-HT1A receptors (pKi 9.74 in pig cortex, pD2 6.82 for contraction of rabbit basilar artery) and a moderate to low affinity at histamine H1-receptors (pA2 6.74) and beta1-ARs (pA2 5.75), B8805-033 did not interact with a number of other neurotransmitter receptors (pKi or pA2<5.0). From the i.v. doses of B8805-033 to either inhibit the urethral pressure response to NA by 50% (29 nmol/kg) or to evoke a fall in diastolic blood pressure by 25% (1.54 micromol/kg) in anaesthetized dogs, an urethral/ vascular selectivity ratio of 52 was obtained, far exceeding that found for the nearly unselective prazosin (ratio 1.8). We conclude that B8805-033 is a highly alpha1A-AR selective antagonist, which may potentially be useful as pharmacological tool to investigate alpha1-AR heterogeneity and in the treatment of benign prostatic hyperplasia. PMID- 11414661 TI - Involvement of adenosine in the effect of antidepressants on glutamate and aspartate release in the rat prefrontal cortex. AB - The effect of local administration of amitriptyline (AMI), desipramine (DMI) and citalopram (CIT) on veratridine-evoked glutamate (Glu) and aspartate (Asp) release in the prefrontal cortex of the conscious rat was examined using in vivo reverse microdialysis. The antidepressants (each at 100 microM) significantly reduced Glu and Asp release. The effect of AMI and CIT was attenuated by i.p. administration of the adenosine A1/A2A receptor antagonist caffeine (10 mg/kg), or by local infusion of the adenosine A1 receptor antagonist 8 cyclopentyltheophylline (CPT, 75 microM). Neither caffeine nor CPT influenced the effect of DMI (100 microM). The inhibitory action of DMI at a lower concentration (50 microM) was diminished significantly by CPT, but not caffeine. Perfusion of 5 hydroxytryptamine (5-HT; 100 microM) and the selective agonist of adenosine A1 receptors N6-cyclopentyladenosine (CPA; 50 and 100 microM) also suppressed Glu and Asp release. It is suggested that the blockade of the cellular uptake of adenosine, or indirect enhancement of its release, and subsequent activation of adenosine A1 receptors may be responsible for the inhibitory effect of antidepressants on Glu and Asp release. PMID- 11414662 TI - Agmatine and putrescine uptake in the human glioma cell line SK-MG-1. AB - The pharmacological properties of a specific agmatine uptake mechanism were investigated in the human glioma cell line SK-MG-1 and compared with those of the putrescine transporter expressed by the same cells and with those of several other organic cation transport systems or ion channels reported in the literature. The specific accumulation of [14C]agmatine at 37 degrees C above nonspecific accumulation at 4 degrees C was energy-dependent and saturable with a Vmax of 64.3+/-3.5 nmol/min per mg protein and a Km of 8.6+/-1.4 microM. Specific accumulation was attenuated by replacement of extracellular Na+ by choline by 65%, not affected by lithium and enhanced by replacement by sucrose. Phentolamine, clonidine, 1,3-di(2-tolyl)guanidine, histamine, putrescine, spermine and spermidine were inhibitors of specific [14C]agmatine accumulation. In contrast, corticosterone, desipramine, O-methylisoprenaline, cirazoline, moxonidine, L-arginine, L-lysine, verapamil, nifedipine and CdCl2 at concentrations up to 10 mM failed to inhibit specific [14C]agmatine accumulation, thus excluding that the latter is mediated by amino acid or monoamine carriers, by Ca2+ channels or by the organic cation transporters OCT1, OCT2, OCT3, OCTN1 or OCTN2. The pattern of activity of inhibitory compounds was also different from that determined for specific putrescine accumulation found in the same cells (Km 1.3+/-0.1 microM, Vmax 26.1+/-0.4 nmol/min per mg protein) ruling out an identity of the specific [14C]agmatine and [14C]putrescine accumulation mechanisms. It is concluded that specific accumulation of agmatine in human glioma cells is mediated by a specific transporter whose pharmacological properties are not identical to those of the agmatine transporter previously identified in rat brain synaptosomes and to other so far known carrier mechanisms for organic cations and ion channels. The agmatine uptake system may be important for the regulation of the extracellular concentration of agmatine in man. PMID- 11414663 TI - Modulation of potassium current and calcium influx by somatostatin in rod bipolar cells isolated from the rabbit retina via sst2 receptors. AB - Somatostatin (somatotropin release-inhibiting factor, SRIF) receptor subtypes are expressed by several retinal neurons, suggesting that SRIF acts at multiple levels of the retinal circuitry, although functional data on this issue are scarce. Of the SRIF receptors, the sst2A isoform is expressed by rod bipolar cells (RBCs) of the rabbit retina, and in isolated RBCs we studied the role of sst2 receptors in modulating both K+ current (IK) and the intracellular free [Ca2+] ([Ca2+]i) using both voltage-clamp and Ca2+-imaging techniques. SRIF and octreotide (a SRIF agonist that binds to sst2 receptors) inhibited that component of IK corresponding to the activation of large-conductance, Ca2+- and voltage dependent K+ channels (IBK) and reduced the K+-induced [Ca2+]i accumulation, suggesting that SRIF effects on IBK may have been secondary to inhibition of Ca2+ channels. Octreotide effects on IBK or on [Ca2+]i accumulation were prevented by RBC treatment with L-Tyr8-Cyanamid 154806, a novel sst2 receptor antagonist, indicating that SRIF effects were mediated by sst2 receptor activation. The present data indicate that SRIF may modulate the information flow through second order retinal neurons via an action predominantly at sst2 receptors, contribute to the proposition that SRIF be added to the growing list of retinal neuromodulators, and suggest that one of its possible roles in the retina is to regulate transmitter release from RBCs. PMID- 11414664 TI - Reproducibility of ultrasound blood flow measurement of the superior mesenteric artery before and after exercise. AB - This study examines the reproducibility of gastro-intestinal blood flow measurements in the superior mesenteric artery (SMA) both before and immediately after exercise with Doppler ultrasound measurements. Twelve well-trained males (mean +/- SD: age 25.9 +/- 3.8 yr; VO2max 4.8 +/- 0.91 x min(-1)) were measured twice (trial 1 and 2) with a 1 week interval before and immediately after 1 hr cycling at 70% VO2max. Duplex scanning was performed with the athletes in supine position immediately after transition from a chair (before exercise) or bicycle (after exercise). The variability of three measurements before exercise was studied within both trials (short-term reproducibility) and the mean pre-exercise values were compared between the trials (long-term reproducibility). In addition, post-exercise measurements were compared in the same way. Reproducibility was tested using the coefficient of variation and Cronbach's alpha. Mean pre-exercise blood flow was 424 +/- 66 ml/min (n = 12) in trial 1 and 375 +/- 38 ml/min (n = 11) in trial 2. Immediately after exercise blood flow had decreased by 49% to 214 +/- 36 ml/min (p <0.01) in trial 1 and by 38% to 234 +/- 36 ml/min (p < 0.01) in trial 2. Blood flow before and after exercise was not significantly different between trials (paired t-test) and therefore reproducible at the group level. Before exercise a good to fair reproducibility was observed both at the short term (Cronbach's alpha: 0.88 in trial 1, 0.73 in trial 2, n = 11), and at the long-term (alpha = 0.80, n= 11). In contrast, long-term reproducibility immediately after exercise was poor (alpha = -0.99, n = 8 and alpha = 0.36, n = 7 after the first and second cycling period, respectively). In conclusion, duplex scanning of SMA after a sitting-supine transition in well-trained subjects is not a reproducible method at the individual level for intestinal blood flow measurements immediately after exercise. PMID- 11414665 TI - Genes and olympic performance: a co-twin study. AB - An Olympic gold medalist in a 20 km competitive walking race and his identical twin brother, also an Olympic athlete in the same event but with inferior performance, were tested in order to obtain some further insight into the relative importance of genetic factors in modulating athletic excellence. Both twins had undergone the same strenuous, long-term training for 19 years since the age of 15 under the guidance of the same coach. An assessment of their bio behavioural profiles at 40 years of age, i. e. 7 years after they ceased training, revealed that intrapair differences were negligible in physiological attributes but divergent in personality traits measured. Respective values for the Olympic winner and his identical counterpart were as follows: Body mass index 23.2 and 22.7, cardiac mass index 85.4 and 84.4 g x m2, squatjumping 25.3 and 27.3 cm, VO2 at running speed 9 km x h(-1) 33.1 and 33.6 ml x kg(-1) x min(-1), VO2 max 57.1 and 58.6 ml x kg(-1) x min(-1) (72.5ml x kg(-1) x min(-1) for the Olympic winner at age 22 yrs), reaction to anger 97 and 9 and anger expression 2 and 76 in percentile of the State-Trait Anger Expression Inventory. Findings suggest that although genetic constitution and years of physical training are prerequisites for making an Olympic athlete, success may be largely influenced by personality traits. PMID- 11414666 TI - N-acetylcysteine attenuates oxidative burst by neutrophils in response to ergometer rowing with no effect on pulmonary gas exchange. AB - This study evaluated whether the reduction of the neutrophil oxidative burst by N acetylcysteine improves pulmonary gas exchange during a six minute maximal ergometer row. Healthy trained oarsmen were double-blinded randomized to either N acetylcysteine (6 g daily for three days) or placebo groups. As determined by the relative changes of the zymosan-stimulated luminol-enhanced chemiluminescence response, N-acetylcysteine suppressed the exercise-induced enhanced neutrophil oxidative burst response to rowing (-7 +/- 6% vs. 17 +/- 8%; P < 0.05). This was the case although the concentration of neutrophils remained similarly elevated above the pre-exercise level in both trials (to 5.4+/-0.5 vs. 5.9+/-0.6 x 10(9) cells x l(-1), respectively, P>0.05). In the placebo and N-acetylcysteine groups, pulmonary ventilation increased and the arterial CO2 partial pressure decreased to the same extent during exercise. Also, at the end of exercise the arterial O2 partial pressure (77 1 vs. 78+/-1 mmHg), haemoglobin O2 saturation (92 +/- 1% vs. 93 +/- 1%) and O2 uptake (5.0 +/- 0.2 vs. 4.9 +/- 0.21 x min(-1)) were not significantly affected by N-acetylcysteine. Equally, two hours after exercise, the pulmonary diffusion capacity was reduced by 7 +/- 2% below the pre-exercise with no significant influence of N-acetylcysteine. We conclude that the neutrophil oxidative burst to exercise does not influence pulmonary gas exchange during and after maximal rowing. PMID- 11414667 TI - Tissue expression and plasma concentrations of TNFalpha, IL-1beta, and IL-6 following treadmill exercise in mice. AB - Exercise can increase plasma inflammatory cytokine concentrations in humans, but tissue responses are not well studied. We examined plasma concentrations and tissue expression of TNFalpha, IL-1beta, and IL-6 following treadmill running in mice. C57B1/6 mice were randomly assigned to: non-exercise control (CON), sacrifice at 0 or 1.5 h after 60 min running (MOD0, MOD 1.5), sacrifice at 0, 1.5, or 3 h after fatiguing running (approximately 3 h) (EX0, EX1.5, EX3), or lipopolysaccharide (25 microg) with no exercise (LPS). Lung, liver, muscle, and brain mRNA expression was analyzed (n = 4-6/group) using reverse transcriptase rapid polymerase chain reaction (RT-RPCR). Plasma cytokine concentrations were determined (n =4-10/group) by ELISA. Plasma IL-6 was higher in EX1.5, and lung TNFalpha mRNA was higher in EX1.5 and EX3 compared to CON (P < 0.05). No significant increases in plasma cytokine concentrations or tissue cytokine expression were found in other EX groups. LPS significantly increased these cytokine measures in tissues and plasma, with the exception of plasma IL-1beta which was undetectable. The source of the plasma IL-6 following exercise does not appear to be lung, liver, muscle, or brain tissue, and remains to be determined. These data also suggest that tissue level cytokine expression may not necessarily lead to increased plasma cytokine concentrations. PMID- 11414668 TI - Respiratory arrest in a male athlete after running through a wheat field. AB - A male athlete suffered complete respiratory arrest after collapsing at the entrance to an Accident and Emergency Department of a hospital. He was resuscitated and recovered fully after several hours. He had multiple wheals on both legs and had complained of severe breathlessness before collapsing. The athlete had run through a wheat field, upon finding his usual pathway overgrown, which had caused wheat pollen to be released. He had been a mild asthmatic for 30 years and had recalled some mild urticaria after exercise but had never experienced exercise-induced asthma, or hay fever. Subsequent allergy tests showed negative for wheat but positive for house dust mite and grass mix. It is possible that the symptoms were triggered either by the running itself, inhalation of allergens other than wheat pollen, skin abrasions caused by contact with wheat stalks, or a combination of these factors. PMID- 11414669 TI - Reliability of mean power recorded during indoor and outdoor self-paced 40 km cycling time-trials. AB - The purpose of this study was to assess reliability of both indoor and outdoor 40 km time-trial cycling performance. Eight trained cyclists completed three indoor 40 km time-trials on an air-braked ergometer (Kingcycle) and three outdoor 40 km time-trials on a local course. Power output was measured for all trials using the SRM powermeter. Mean performance time across three indoor trials was 54.21 +/- 2.59 (min:sec) and was significantly different (P<0.05) to mean time across three outdoor trials (57.29 +/- 3.22 min:sec). However, there was no significant difference (P = 0.34) for mean power across three indoor trials (303+/-35W) when compared to outdoor performances (312 +/- 23 W). Within-subject variation for mean power output expressed as a coefficient of variation (CV) improved in both indoors and outdoors for trials 2 and 3 (CV = 1.9%, 95% CI 1.0 - 3.4 and CV = 2.1 %, 95 % CI 1.1 - 3.8) when compared to trials 1 and 2 (CV=2.1%, 95% CI 1.2-3.8 and CV=2.4%, 95% CI 1.3-4.3). These findings indicate that power output measured using the SRM powermeter is highly reproducible for both laboratory-based and actual 40 km time-trial cycling performance. PMID- 11414670 TI - Long term changes of the throwing arm of former elite javelin throwers. AB - The aim of this study was to determine long term changes in shoulder and elbow joints of former elite javelin throwers. Twenty-one elite javelin throwers were examined at an average of 19 years after the end of their high performance phase. Mean age at examination was 50 years. Functional assessment of both shoulders was determined by the Constant-score. The shoulder of the throwing arm was examined by magnetic resonance imaging. Both elbow joints were examined clinically and radiographically. Five athletes complained about transient shoulder pain in their throwing arm affecting activities of daily living, fourteen athletes had a deficit of internal rotation of at least ten degrees. Constant-scores of throwing arms were six points lower than those of non-throwing arms (P < 0.05). Complete ruptures and partial tears of the rotator cuff were frequent. Three athletes complained about transient elbow pain in their throwing arm affecting activities of daily living; ten athletes had a deficit of extension of more than five degrees. All dominant elbows had advanced arthrotic alterations (osteophytes, sclerosis) compared to the non-dominant side. Athletes who trained with weights of more than 3 kg had a significantly higher risk of degenerative changes than athletes who did not (P < 0.01). We therefore recommend to avoid throwing training with weights of more than 3 kg. PMID- 11414671 TI - Validation of the COSMED K4 b2 portable metabolic system. AB - The purpose of this investigation was to assess the accuracy of the COSMED K4 b2 portable metabolic measurement system against the criterion Douglas bag (DB) method. During cycle ergometry on consecutive days, oxygen consumption (VO2), carbon dioxide production (VCO2), minute ventilation (VE), and respiratory exchange ratio (R) were measured at rest and during power outputs of 50, 100, 150, 200, and 250W. No significant differences (P > 0.05) were observed in VO2 between the K4 b2 and DB at rest and at 250W. Though the K4 b2 values were significantly higher (P<0.05) than DB values at 50, 100, 150, and 200 W, the magnitude of these differences was small (0.088, 0.092, 0.096, and 0.088 L x min( 1), respectively). VCO2 and VE values from the K4 b2 were significantly lower than the DB at 200 and 250 W, while no significant differences were observed from rest through 150W. The slight overestimation of VO2 (50-200 W) combined with the underestimation of VCO2 (200 and 250W) by the K4 b2 resulted in significantly lower R values at every stage. These findings suggest the COSMED K4 b2 portable metabolic measurement system is acceptable for measuring oxygen uptake over a fairly wide range of exercise intensities. PMID- 11414672 TI - Gain in strength and muscular balance after balance training. AB - The isolated effect of balance training on muscle strength of the flexors and extensors of the knee, without accompanying strength training, has not been addressed in the past. Effects of a balance training program alone were compared to a strength training program. Balance and strength training were performed by 15 persons each for 6 weeks including 12 training units of 25 min. Balance training was performed on instability training devices such as rolling board, mini trampoline and large rubber ball. The 15 persons of the strength training group trained on machines for leg curls and on leg presses for 25 min per unit. Measurements for balance were performed with one-leg balance on a narrow edge and a tilting stabilometer for 30 s; maximum isometric strength was measured using an isokinetic device for each leg separately. The muscular balance between dominant and non-dominant leg was calculated. Strength gain was similar for the flexors and extensors in both groups. One-leg balance improved after balance training (P< 0.01) with a 100% increase over the strength training group (P < 0.05) and the stabilometer test for each person in the balance (P < 0.01), but not in the strength training group. In the balance group the initial difference between right and left diminished. The results indicate balance training to be effective for gain in muscular strength, and secondly, in contrast to strength training, equalisation of muscular imbalances may be achieved after balance training. PMID- 11414673 TI - Validation of a computerized metabolic measurement system (Oxycon-Pro) during low and high intensity exercise. AB - The aim of the study was to validate a new, fast metabolic measurement system (the Oxycon-Pro) during low and high exercise intensities against the Douglas bag method. For this purpose twelve highly trained subjects performed an incremental cycle ergometer test. In the third minute of each exercise step simultaneous measurements of the Douglas bag and the Oxycon-Pro were performed. No significant differences between the Oxycon-Pro and the Douglas bag measurements for minute ventilation (VE), oxygen uptake (VO2) and carbon dioxide expiration (VCO2) were found. Bland and Altman analysis of validity demonstrated minimal bias and low standard deviations. In conclusion, the results show that the Oxycon-Pro is a valid apparatus for determination of minute ventilation (VE), oxygen uptake (VO2) and carbon dioxide expiration (VCO2) during low as well as during maximal exercise intensities. Unlike with the Douglas bag method, this fast metabolic measurement system can be used for accurate and quick determination of ventilatory variables during exercise. PMID- 11414674 TI - High-intensity aerobic training during a 10 week one-hour physical education cycle: effects on physical fitness of adolescents aged 11 to 16. AB - The aim of this study was to analyse the effects of a high-intensity aerobic training program on different components of physical fitness in adolescents aged 11 to 16 years. The subjects were divided into a high intensity (HI) group (243 girls and 260 boys) and a control (C) group (21 girls and 27 boys). HI and C completed a weekly 3 hour physical education (PE) session. Before and after a 10 week period, the two groups performed the European physical fitness test battery (EUROFIT). During these 10 weeks HI spent one hour out of three at a specific PE session. These specific sessions consisted of short intermittent exercises (10 seconds) at 100 to 120% of maximal aerobic speed. They showed a significant influence on standing broad jump (2.9 %, P<0.05, F=4.85), 20 meter shuttle run (3.8%, p0.001, F=23.21) and on the maximal distance covered over 7 min (7.6 %, P< 0.001, F= 14.48). For C there was no improvement in EUROFIT performances. It was concluded that training at high intensity improves not only children's aerobic fitness but also performance of standing broad jump. Well-monitored, adequate intensive training is necessary for a more desirable functional development. PMID- 11414675 TI - Change in strength and dexterity after open carpal tunnel release. AB - Few studies have documented the long-term recovery of hand function after open carpal tunnel release surgery. Grip strength and manual dexterity were measured before and every five weeks up to 25 weeks after open carpal tunnel release in a sample of 11 affected wrists. Dexterity scores prior to surgery were substantially (nearly 3sd) below normative levels. There was a significant (t10=5.45, P<0.001) 37% decrease in grip strength immediately following surgery. Trend analysis of the data indicated a significant second order pattern for the decrease and recovery of grip strength, and a significant linear pattern of recovery of dexterity following surgery. The strength and dexterity tests were effective in documenting changes in function in recovery from open carpal tunnel release surgery, but grip strength and dexterity in these subjects only recovered to pre-operative levels at 25 weeks. PMID- 11414676 TI - Long-term outcome of operative or nonoperative treatment of anterior cruciate ligament rupture--is sports activity a determining variable? AB - The purpose of this study was to evaluate the long-term clinical outcome of operative versus nonoperative treatment of anterior cruciate ligament (ACL) deficiency and to define its relationship with sports activity. Forty-six patients (37 male, 9 female, mean age, 33.6 +/- 8.0 years) who underwent open ACL reconstruction using patellar tendon autograft and 25 patients (18 male, 7 female, mean age, 32.3 +/- 9.9 years) who were treated nonoperatively were evaluated by the same two examiners at 5 - 7 and 10 - 13 years following injury. The evaluations included objective and subjective scoring (Lysholm, OAK, IKDC), instrumented testing (Cybex, KT 1000), radiographic evaluation, and assessments of sports activity, with respect to type, frequency and associated symptoms. According to Lysholm, OAK and IKDC scores, the operative group performed significantly (p < 0.05) better and was able to maintain increased involvement in sports, although both groups participated less over time. However, risks for degenerative joint changes were similar for both the operative and the nonoperative group. A significant (p < 0.05) correlation between participation in high-risk pivoting sports, such as soccer or basketball and osteoarthritic changes could be found for the nonoperative group, only. Sports activity represents an important variable not only affecting the outcome, but also influencing treatment decisions following ACL injuries. PMID- 11414677 TI - Short-term dietary energy restriction reduces lean body mass but not performance in physically active men and women. AB - We studied the effect of moderate, short-term energy restriction on physical performance in physically fit men (n = 13) and women (n = 11) in a controlled clinical research setting with a metabolic kitchen, exercise testing laboratory and training facility. The experiment consisted of a 10 d baseline period followed by either 2 wk of dietary energy restriction (750 kcal/d; n = 16) or energy balance (control; n = 8). During this 24 day study, exercise energy expenditure averaged 465 +/- 5.7 kcal/d in all subjects and was accomplished through treadmill running at a self-selected pace. Body weight was maintained in the control group (-0.36 +/- 0.24kg), but energy restriction resulted in weight loss of -1.29 +/- 0.16 kg (p < 0.001). There was a trend for lean body mass to decline more in the energy restriction group (p = 0.093), accounting for 61% of the weight loss, and urinary nitrogen excretion also tended to be higher in the energy restriction vs. control group (i.e., 13.2 +/- 1.1 vs. 11.2 +/- 1.0g/d; p = 0.089). Muscle strength (leg & shoulder press; 1 repetition maximum) was maintained or increased during the energy restriction period. Muscle endurance, assessed by leg squats to fatigue, and 5 mile run time improved following two weeks of energy restriction or balance. Anaerobic capacity (Wingate Test) increased slightly in the restriction (+ 368 +/- 219 joules) but declined in the control group 649 +/- 288 joules; p<0.05). We conclude that short-term (2 weeks) moderate energy restriction (approximately 750 kcal/d) results in weight loss but does not impair performance in physically fit young men and women. PMID- 11414678 TI - Induction of nitric oxide synthase during Japanese encephalitis virus infection: evidence of protective role. AB - The ability of Japanese encephalitis virus (JEV) and JEV-induced macrophage derived factor (MDF) to modulate nitric oxide synthase (NOS) activity in brain and tumor necrosis factor-alpha (TNF-alpha) and the possible antiviral role of NOS during JEV infection were investigated. NOS activity and particularly that of the inducible form of NOS (iNOS) was significantly enhanced in JEV or JEV-induced MDF-treated mice. Following JEV infection, total NOS activity in brain was gradually increased from Day 3 and reached a peak on Day 6. MDF-induced NOS activity and iNOS activity were dose dependent and maximum activity was observed at 1 h after treatment. The response was sensitive to anti-MDF antibody treatment and N(G)-monomethyl-L-arginine (L-NMMA), an inhibitor of NOS. Pretreatment of JEV infected mice with L-NMMA increased the mortality as evident from reduced mean survival time (MST, 11.8 days) compared to placebo treated JEV-infected mice (MST, 17 days). The enhanced level of TNF-alpha observed in the early phase of JEV infection correlated well with the enhanced activity of iNOS. These observations thus provide evidence of the protective role of iNOS during JEV infection and indicate that iNOS may be a key mediator in host innate immune response to infection. PMID- 11414679 TI - Fatty acid delta(5)-desaturase mRNA is regulated by dietary vitamin A and exogenous retinoic acid in liver of adult rats. AB - Delta(5)-Desaturase (D5D) catalyzes the Delta(5,6) desaturation of dietary essential fatty acids of the n-6 and n-3 series. By subtraction hybridization of vitamin A (VA)-deficient and control rat liver cDNA libraries, we isolated a 106 bp cDNA fragment that proved to be homologous to human liver D5D cDNA and used it as a probe to analyze rat D5D mRNA and clone the rat full-length cDNA. Delta(5) Desaturase mRNA was threefold more abundant in liver from VA-deficient rats than in liver from VA-sufficient rats and was expressed dose dependently when dietary VA was varied (VA marginal > control > VA supplemented). Treatment of VA deficient rats with all-trans-retinoic acid lowered the level of expression of D5D mRNA toward that of VA-sufficient rats. The 3413-bp full-length D5D cDNA cloned from rat liver contains an open reading frame of 447 amino acid residues sharing 92% similarity with its human counterpart. Expression of this cDNA in HEK293T cells incubated with dihomo-gamma-linolenic acid (20:3, n-6) resulted in a significantly increased ratio of the product, arachidonic acid (20:4, n-6), to substrate in cell lipid extracts. Delta(5)-Desaturase mRNA is expressed in relatively high abundance in rat adrenal gland and mammary tissue and moderately in liver, kidney, lung, spleen, thymus, brain, and eye. The regulation of D5D by VA could be important for growth and development, and reproduction, as well as in the control of inflammation. PMID- 11414680 TI - A Mg-dependent ecto-ATPase in Leishmania amazonensis and its possible role in adenosine acquisition and virulence. AB - The plasma membrane of cells contains enzymes whose active sites face the external medium rather than the cytoplasm. The activities of these enzymes, referred to as ectoenzymes, can be measured using living cells. In this work we describe the ability of living promastigotes of Leishmania amazonensis to hydrolyze extracellular ATP. In these intact parasites whose viability was assessed before and after the reactions by motility and by trypan blue dye exclusion, there was a low level of ATP hydrolysis in the absence of any divalent metal (5.39 +/- 0.71 nmol P(i)/h x 10(7) cells). The ATP hydrolysis was stimulated by MgCl(2) and the Mg-dependent ecto-ATPase activity was 30.75 +/- 2.64 nmol P(i)/h x 10(7) cells. The Mg-dependent ecto-ATPase activity was linear with cell density and with time for at least 60 min. The addition of MgCl(2) to extracellular medium increased the ecto-ATPase activity in a dose-dependent manner. At 5 mM ATP, half-maximal stimulation of ATP hydrolysis was obtained with 1.21 mM MgCl(2). This stimulatory activity was also observed when MgCl(2) was replaced by MnCl(2), but not by CaCl(2) or SrCl(2). The apparent K(m) for Mg ATP(2-) was 0.98 mM and free Mg(2+) did not increase the ecto-ATPase activity. In the pH range from 6.8 to 8.4, in which the cells were viable, the acid phosphatase activity decreased, while the Mg(2+)-dependent ATPase activity increased. This ecto-ATPase activity was insensitive to inhibitors of other ATPase and phosphatase activities, such as oligomycin, sodium azide, bafilomycin A(1), ouabain, furosemide, vanadate, molybdate, sodium fluoride, tartrate, and levamizole. To confirm that this Mg-dependent ATPase was an ecto-ATPase, we used an impermeant inhibitor, 4,4'-diisothiocyanostylbene 2',2'-disulfonic acid as well as suramin, an antagonist of P(2) purinoreceptors and inhibitor of some ecto ATPases. These two reagents inhibited the Mg(2+)-dependent ATPase activity in a dose-dependent manner. A comparison between the Mg(2+)-dependent ATPase activity of virulent and avirulent promastigotes showed that avirulent promastigotes were less efficient than the virulent promastigotes in hydrolyzing ATP. PMID- 11414681 TI - Topology of the C-terminus of sodium hydrogen exchanger isoform-1: presence of an extracellular epitope. AB - In this study, the topology of the C-terminus of Na(+)/H(+) exchanger-1 (NHE-1) was examined using red blood cell (RBC) membrane vesicles. Specific polyclonal antibodies were raised against the C-terminus of the rat NHE-1 using a glutathione S-transferase fusion protein antigen. Enzyme-linked immunosorbent assay (ELISA) and flow cytometer analyses were employed. NHE-1 antibodies recognized a 110-kDa protein in the rats and mice, but not in the humans. RBC vesicles were resealed using 1 mM MgCl(2), and the right side out (RSOVs) and the inside out vesicles (ISOVs) were selected by concanavalin A. NHE-1 antibodies reacted strongly with the RSOVs and poorly with the ISOVs in an ELISA. The reaction with the unsealed vesicles was stronger than the resealed RSOVs. In contrast, the monoclonal alpha-actin antibodies reacted poorly with the RSOVs, but extremely strongly with the ISOVs. Flow cytometer analysis also showed a strong reaction of NHE-1 antibodies with the intact and permeabilized rat RBCs. The antibodies raised in this study also contain a population which reacts with the internal epitope in the NHE-1 C-terminus. These findings suggest that the NHE 1 C-terminus contains epitope(s) exposed extracellularly. PMID- 11414682 TI - Cloning, sequencing, tissue distribution, and heterologous expression of rat flavin-containing monooxygenase 3. AB - The sequence of rat FMO3 was obtained by RT-PCR and 5'/3' terminal extension. Complete cDNA was amplified, cloned, and sequenced. The cDNA encodes a protein of 531 amino acids which contains the NADPH- and FAD-binding sites and a hydrophobic carboxyl terminus characteristic of FMOs. This sequence is 81, 81, and 91% identical to sequences of human, rabbit, and mouse FMO3, respectively, and 60% identical to rat FMO1. Rat FMO3 was expressed in Escherichia coli. The recombinant protein and the native protein purified from rat liver microsomes migrated with the same mobility (56 kDa) as determined in sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting. Recombinant rat FMO3 showed activities of methimazole S-oxidation, and NADPH oxidation associated with the N- or S-oxidation of trimethylamine and thioacetamide, in good concordance with those reported for human FMO3. When probed with rat FMO3 cDNA (bases 201 to 768), a strong signal corresponding to the 2.3-kb FMO3 transcript was detected in RNA samples from rat liver and kidney while a weak signal was observed with lung RNA samples. In contrast, the probe did not hybridize with any RNA from brain, adipose tissue, or muscle. PMID- 11414683 TI - Effect of STAT5b on rat liver alcohol dehydrogenase. AB - Growth hormone (GH) enhances rat liver alcohol dehydrogenase (ADH) due to an increase in enzyme synthesis, which is mediated at the level of transcription. Previous studies have shown that the effect of GH in enhancing activation of the ADH promoter is mediated by C/EBP beta binding to region -22 to -11 relative to the start of transcription. In this study, STAT5b and C/EBP beta were found to bind to adjacent nucleotide sequences on a region between -226 and -194. Expression vectors for both STAT5b and C/EBP beta independently activated the promoter. Furthermore, the expression vector for the GH receptor also activated the ADH promoter, and this effect was abrogated by mutations of the adjacent STAT5b and C/EBP beta binding sites. These observations indicate that the enhancing effect of GH is mediated by both STAT5b and C/EBP beta. PMID- 11414684 TI - Covalent alteration of the CYP3A4 active site: evidence for multiple substrate binding domains. AB - The inhibition of CYP3A4-mediated oxidation of triazolam and testosterone was assessed in the presence of a selection of known CYP3A4 substrates and inhibitors. Under experimental conditions where the Michaelis-Menten model predicts substrate-independent inhibition ([S] = K(m)), results yielded substrate dependent inhibition. Moreover, when the same experimental design was extended to a group of structurally similar flavonoids it was observed that flavanone, flavone, 3-hydroxyflavone, and 6-hydroxyflavone (10 microM) activated triazolam metabolism, but inhibited testosterone hydroxylation. In additional studies, residual CYP3A4 activity toward testosterone and triazolam hydroxylation was measured after pretreatment with the CYP3A4 mechanism based inhibitor, midazolam. After midazolam preincubation, CYP3A4 6 beta-hydroxylase activity was reduced by 47% while, in contrast, triazolam hydroxylation was reduced by 75%. These results provide physical evidence, which supports the hypothesis that the active site of CYP3A4 contains spatially distinct substrate-binding domains within the enzyme active site. PMID- 11414685 TI - Modulation of the susceptibility of human erythrocytes to snake venom myotoxic phospholipases A(2): role of negatively charged phospholipids as potential membrane binding sites. AB - Cerrophidion (Bothrops) godmani myotoxins I (CGMT-I) and II (CGMT-II), Asp-49 and Lys-49 phospholipases A(2) (PLA2s), which drastically differ in enzymatic activity, were devoid of direct hemolytic effects on erythrocytes (RBC) from different species despite the fact that enzymatically active CGMT-I was able to hydrolyze RBC membrane phospholipids and disrupt liposomes prepared from RBC lipids. Human RBC did not become susceptible to the toxins after treatment with neuraminidase or after altering membrane fluidity with cholesterol or sublytic concentrations of detergent. Unlike normal RBC, significant hemolysis was induced by CGMT-II and another similar Lys-49 isoform, B. asper MT-II (BAMT-II), in RBC enriched with phosphatidylserine (PS). Hemolysis was greater in RBC preincubated with pyridyldithioethylamine (PDA), a potent inhibitor of aminophospholipid transport. RBC enriched with phosphatidic acid (PA) also became susceptible to the myotoxins but was unaffected by PDA. Cells enriched with phosphatidylcholine (PC) remained resistant to the action of the toxins. BAMT-II also induced damage in black lipid membranes prepared with PS but not PC alone. When RBC binding of BAMT-II was measured by enzyme-linked immunosorbent assay, it was observed that PS- and PA-enriched erythrocytes were always able to capture more toxin than normal and PC-enriched RBC. This effect was significantly improved by PDA (in the case of PS) and it was observed either in the presence or in the absence of calcium in the medium. These data suggest that negatively charged lipids in the outer leaflet of cell membranes constitute myotoxic PLA2 binding sites. The scarcity of anionic phospholipids in the outer leaflet of RBC could explain their resistance to the action of these PLA2s. PMID- 11414686 TI - The glucuronyl C5-epimerase activity is the limiting factor in the dermatan sulfate biosynthesis. AB - An early step in the biosynthesis of dermatan sulfate is polymerization to chondroitin, which then is modified by the D-glucuronyl C5-epimerase and mainly 4 O-sulfotransferase. The final structure of the dermatan sulfate side chains varies and our aim was to identify, which of the two enzymes that are crucial to generate dermatan sulfate copolymeric structures in tissues. Dermatan sulfate side chains of biglycan and decorin were prepared from fibroblasts and nasal and articular chondrocytes and characterized regarding detailed structure. Microsomes were prepared from these cells and the activities of D-glucuronyl C5-epimerase and 4-O-sulfotransferase were determined. Chondrocytes from nasal cartilage synthesized biglycan and decorin containing 10%, articular chondrocytes 20--30%, and fibroblast 80% of the uronosyl residues in the l-iduronyl configuration. All three tissues contained high amount of 4-O-sulfotransferase activity. The activity of d-glucuronyl C5-epimerase showed different relationships. Fibroblasts contained a high level of the epimerase activity, articular chondrocytes intermediary activity, and in nasal cartilage it was barely detectable. The data indicate that the activity of the d-glucuronyl C5-epimerase is the main factor for formation of dermatan sulfate in tissues. PMID- 11414687 TI - Quercetin inhibits matrix metalloproteinase-1 expression in human vascular endothelial cells through extracellular signal-regulated kinase. AB - Studies have shown that intake of quercetin was inversely associated with mortality from coronary heart disease. Since recent studies documented that disruption of atherosclerotic plaques is the key event triggering acute myocardial infarction, and vascular endothelium-derived matrix metalloproteinase 1 (MMP-1) contributes to plaque destabilization, we examined the effect of quercetin on MMP-1 expression in human vascular endothelial cells. Our results showed that quercetin significantly inhibited basal and oxidized LDL (oxLDL) stimulated MMP-1 expression. Our data also indicated that extracellular signal regulated kinase (ERK) mediated the basal and oxLDL-stimulated expression of MMP 1, and quercetin is a potent inhibitor of ERK, suggesting that quercetin may inhibit MMP-1 expression by blocking the ERK pathway. Finally, we showed that quercetin stimulated tissue inhibitor of metalloproteinase-1 expression in oxLDL- and PMA-treated cells. In conclusion, the present study demonstrated for the first time that quercetin inhibited MMP-1 expression in vascular endothelial cells, suggesting that quercetin might contribute to plaque stabilization. PMID- 11414688 TI - Efficiency and mechanism of the antioxidant action of trans-resveratrol and its analogues in the radical liposome oxidation. AB - trans-Resveratrol (trans-3,5,4'-trihydroxystilbene) is a nonflavonoid polyphenol reported to exert different biological activities, among them inhibition of the lipid peroxidation, scavenging of the free radicals, inhibition of the platelet aggregation, and anticancer activity as the most important. In order to enlighten the radical-scavenging mechanism of trans-resveratrol, stationary gamma radiolytic experiments in liposomes and pulse radiolytic experiments in aqueous solutions were performed. Applying the stationary gamma-radiolysis together with the subsequent product analysis, reactions of lipid peroxyl radicals, LOO*, with trans-resveratrol and other natural antioxidants were investigated. It was found that trans-resveratrol was a better radical scavenger than vitamins E and C but similar to the flavonoids epicatechin and quercetin. The comparison of the radical-scavenging effects of trans-resveratrol and its analogues trans-4 hydroxystilbene and trans-3,5-dihydroxystilbene revealed that trans-resveratrol and trans-4-hydroxystilbene showed almost the same effect and were more efficient than trans-3,5-dihydroxystilbene. These findings indicate greater radical scavenging activity of trans-resveratrols para-hydroxyl group than its meta hydroxyl groups. Using the pulse radiolysis, reactions of trans-resveratrol and its analogues with trichloromethylperoxyl radicals, CCl(3)OO*, were studied. Spectral and kinetic properties of the observed transients showed great similarity between trans-resveratrol and trans-4-hydroxystilbene which seems to confirm that para-hydroxyl group of trans-resveratrol scavenges free radicals more effectively than its meta-hydroxyl groups. PMID- 11414689 TI - Uptake of the neutral amino acids glutamine, leucine, and serine by Pneumocystis carinii. AB - Experiments to elucidate the mechanism by which Pneumocystis carinii transports glutamine, leucine, and serine were performed in this study. Uptake of all three radiolabeled amino acids exhibited first-order, saturation kinetics as extracellular substrate concentrations were increased, thus ruling out simple diffusion and indicating carrier-mediated transport. Kinetic analyses of amino acid uptake and the results of competitive inhibition experiments suggested that leucine, serine, and glutamine were taken up via a common transporter system. The uptake of serine was examined in greater detail to characterize the nature of the carrier. Serine uptake was not affected by N, N'-dicyclohexylcarbodiimide, carbonyl cyanide m-chlorophenyl hydrazone, ouabain, gramicidin, valinomycin, sodium azide, salicylhydroxamine acid (SHAM), iodoacetate, iodoacetate plus SHAM, KCN, and azide. Thus serine uptake did not require sodium or energy from ATP, an electrochemical proton gradient or a membrane potential across the cell surface (i.e., proton-motive force). Serine uptake was dependent on glucose in the extracellular compartment. In the presence of glucose, serine uptake was inhibited by chloramphenicol but not cycloheximide. The results from these experiments are most consistent with facilitated diffusion as the mechanism. After 30 min of incubation, most of the radioactivity was in the cellular soluble fraction. In most cases, incorporation into the extractable total lipids and the remaining particulate cellular components were detectable after this incubation period. PMID- 11414690 TI - Differential effects of naturally occurring isothiocyanates on the activities of cytochrome P450 2E1 and the mutant P450 2E1 T303A. AB - The effects of benzyl (BITC) and phenethyl isothiocyanate (PEITC) on the activity of a P450 2E1 mutant where the conserved threonine at position 303 was replaced with an alanine residue (P450 2E1 T303A) were examined. PEITC inactivated the mutant enzyme with a K(I) of 1.6 microM. PEITC also inactivated the wild-type P450 2E1 as efficiently with a K(I) of 2.7 microM. The inactivation was entirely dependent on NADPH and followed pseudo-first-order kinetics. Previously we reported the mechanism-based inactivation of wild-type P450 2E1 by BITC with a K(I) of 13 microM. In contrast to the wild-type enzyme, the P450 2E1 T303A mutant was not inactivated by BITC but it was inhibited in a competitive manner with a K(i) of 3 microM. The binding constants determined by spectral binding studies were similar for both enzymes. The binding of BITC produced characteristic Type I spectral changes in the wild-type and mutant enzyme. A radiolabeled BITC metabolite bound to P450 2E1 and to P450 2E1 T303A when both enzymes were incubated with [(14)C]BITC and NADPH. Whole protein electrospray ion trap mass spectrometry indicated that a mass consistent with one molecule of benzylisocyanate and oxygen was adducted to the wild-type enzyme. The mass adducted to the T303A mutant was consistent with the addition of one hydroxylated BITC or of one benzylisocyanate moiety and one sulfur molecule. Analysis of the metabolites of BITC indicated that each enzyme produced similar metabolites but that the mutant enzyme generated significantly higher amounts of benzaldehyde and benzoic acid when compared to the wild-type enzyme. PMID- 11414691 TI - Dynamic features of the subunit interface of Cu,Zn superoxide dismutase as probed by tryptophan phosphorescence. AB - As part of the more general inquiry on the molecular basis of specific recognition between macromolecules, the subunit-subunit interface structure of dimeric superoxide dismutase from Photobacterium leiognathi has been probed selectively by the phosphorescence emission of Trp-73, located at the subunit contact region. Copper at the catalytic site was found to quench completely the delayed emission and therefore all studies were conducted with the copper-free or Cd(2+)-substituted protein. The spectrum at 140 K is diagnostic for an indole ring located in a hydrophobic environment whereas a degree of spectral broadening indicates that the local structure is not unique. Environmental heterogeneity is confirmed by the nonuniform phosphorescence decay in buffer, at 274 K, with lifetime components of 44 and 20 ms of practically equal amplitude. Information on the flexibility of the interface region was gathered from both the intrinsic lifetime and the accessibility of acrylamide to the site of the chromophore. The magnitude of the intrinsic lifetime, its temperature dependence, and the accessibility to solutes like acrylamide describe a tight dimeric structure in which hydrophobic interactions seem to play an important role. In particular the acrylamide bimolecular rate constant is 1.4 x 10(4) M(-1) s(-1) and indicates highly hindered diffusion of the solute through the interface region. Cd(2+) complexation to the apoprotein caused no detectable changes in protein conformation although the metal was able to influence the flexibility of the Trp 73 environment, indicating the occurrence of a long-range communication between the intersubunit surface and the active site, which is more than 16 A away. PMID- 11414692 TI - alpha(2)-Macroglobulin from rheumatoid arthritis synovial fluid: functional analysis defines a role for oxidation in inflammation. AB - A hallmark of inflammation is the release of oxidants, proteinases, and cytokines, all important mediators of the inflammatory cascade. alpha(2) Macroglobulin (alpha(2)M) is a high-affinity, broad-specificity proteinase inhibitor that also binds and regulates the biological activities of a number of cytokines. We demonstrated recently that hypochlorite-oxidized alpha(2)M has decreased ability to inhibit proteinases and regulate cytokines in vitro. The role of oxidation in regulating alpha(2)M functions in vivo is largely unknown. To determine the extent and biological consequence of in vivo alpha(2)M oxidation, we measured the degree of oxidative alpha(2)M modification from rheumatoid arthritis (RA) synovial fluid and compared this with osteoarthritis (OA) as noninflammatory controls. We found that RA synovial fluid alpha(2)M is significantly more oxidized than that from OA. RA synovial fluid also contains a twofold higher median alpha(2)M level than OA, while having only half the alpha(2)M-proteinase inhibitory activity. Detailed biochemical analysis demonstrates proteolytically degraded alpha(2)M in RA greater than in OA synovial fluid. Additionally, the hypochlorite-mediated oxidation product, chlorotyrosine, is present in RA more than in OA or plasma alpha(2)M samples. Taken together, these findings confirm a role for oxidative regulation of inflammation by altering the functions of extracellular mediators such as alpha(2)M. PMID- 11414693 TI - Mouse cyp2g1 gene: promoter structure and tissue-specific expression of a cyp2g1 lacz fusion gene in transgenic mice. AB - The structure of the mouse Cyp2g1 gene was determined to identify regulatory regions important for its olfactory mucosa-specific expression. Two Cyp2g1 genomic clones were isolated and characterized. A 3.6-kilobase 5'-flanking sequence was used to prepare a Cyp2g1--LacZ fusion gene for transgenic mice production. Transgene expression, as determined by beta-galactosidase activity in tissue extracts, was detected in the olfactory mucosa, but not in any other tissues examined, in five different transgenic lines. Thus, the 3.6-kilobase fragment contained regulatory elements sufficient for olfactory mucosa-specific and proper developmental expression of the reporter gene. However, histological and immunohistochemical studies indicated that the expression of the transgene in the olfactory mucosa was patchy and the cellular expression patterns of the transgene did not exactly match that of the endogenous gene. These results implicate the presence of additional regulatory sequences that are necessary for the correct cell type-selectivity within the olfactory mucosa. PMID- 11414694 TI - Structural organization, regulation, and expression of the chloroplastic superoxide dismutase Sod1 gene in maize. AB - A cDNA and genomic clone encoding maize chloroplastic Cu/Zn superoxide dismutase Sod1 were isolated. Southern blot analysis indicated little homology between the chloroplastic (Sod1) and the cytosolic (Sod2, Sod4, Sod4A) cDNAs. Sequence analysis of the genomic clone revealed a promoter, transit peptide, and partial coding sequence. The promoter contained several response elements (e.g., for light, cold temperature, xenobiotics) that may be involved in the regulation of the Sod1 gene. Sod1 expression during development and in response to physiological and chemical stressors such as temperature, xenobiotics (paraquat), and light were examined. PMID- 11414695 TI - Cell differentiation under the influence of rh-BMP-2. AB - Bioactive bone growth factors will likely play an important role in the regeneration of bone. BMP-2 is known to promote osteoblastic cell differentiation and osteogenesis. Whether the BMPs act on human osteoblastic cells by increasing immature cell growth and/or differentiation is unknown. The goal of this study was to analyse possible effects of rhBMP-2 on cell differentiation using a human bone marrow cell culture. rhBMP-2 was added to the culture medium once. Fourteen days after addition of rhBMP-2 the cells were incubated with monoclonal antibodies. The cells were counted and analysed in a fluorescence-activating cell sorter (FACS). Compared to the controls there was an increasing effect on granulocytes, B cells and stem cells. The T-cells and monocytes show no increase or decrease after rhBMP-2 treatment. PMID- 11414696 TI - A role of N-cadherin in neuronal differentiation of embryonic carcinoma P19 cells. AB - N-cadherin is one of the important molecules for cell to cell interaction in the development of the central nervous system (CNS). In this report, we have shown that N-cadherin mRNA and protein were increased rapidly in retinoic acid (RA) induced neuronal differentiation of embryonic carcinoma P19 cells. To explore possible roles for N-cadherin during this process, N-cadherin-overexpressing P19 cell lines were established. These transfected cells could differentiate into neurofilament-expressing neurons in the absence of RA. RT-PCR revealed that the expression patterns of development-related genes, such as Oct-3/4, nestin, Notch 1, and Mash-1 were similar between the transfected P19 cells and the RA-induced wild-type P19 cells during their neuronal differentiation. On the contrary, the Wnt-1 gene was up-regulated in the N-cadherin-overexpressing P19 cells, but could not be detected in the wild-type P19 cells. These results suggest N-cadherin may play a role in neuronal differentiation of P19 cells, possibly through the Wnt-1 signaling pathway. PMID- 11414697 TI - Identification of human ST2 protein in the sera of patients with autoimmune diseases. AB - Soluble human ST2 protein (IL1RL1-a) in the sera of patients with various autoimmune diseases was identified by a newly developed procedure using specific monoclonal antibodies. After immunoprecipitation and subsequent immunoblotting, a glycosylated protein of about 60 kDa was detected in the sera of SLE patients, but not in the sera of healthy controls. The experiments using gel filtration and SDS-PAGE under a nonreducing condition indicated the existence of the ST2 multimer in serum. The mobility of the natural protein was slower than that of the recombinant human ST2 protein produced by COS7 cells in SDS-PAGE, suggesting a difference of glycosylation between humans and monkeys. The identification of the natural human ST2 protein should be important both to fundamental researches and the further clarification of the clinical implications of the ST2 protein. PMID- 11414698 TI - Conservation of death receptor-6 in avian and piscine vertebrates. AB - One of the most recently identified members of the tumor necrosis factor receptor family, death receptor-6 (DR6), has been shown to mediate apoptosis following overexpression in HeLa cells. The avian and piscine orthologs of DR6 have now been identified, and the deduced amino acid sequence for each demonstrates a high level of conservation compared to the mammalian sequence. Expression of dr6 mRNA occurs widely across tissues of both the mature chicken and brook trout. It is now well-established that ovarian follicular atresia occurs via apoptosis originating within the granulosa cell layer. Accordingly, DR6 expression within the ovary was examined to assess the relationship between stage of follicle development and relative levels of this death receptor. Of particular interest was the finding that elevated levels of dr6 mRNA, as well as the translated protein, are expressed in atretic compared to healthy follicles of the hen ovary, thus providing the first association between DR6 expression and apoptosis, in vivo. We conclude that DR6 is a highly conserved and widely expressed death domain-containing receptor and may be implicated in regulating follicle atresia within the vertebrate ovary. PMID- 11414699 TI - Tumor autocrine motility factor is an angiogenic factor hat stimulates endothelial cell motility. AB - Autocrine motility factor (AMF) is a type of tumor-secreted cytokine that primarily stimulates tumor cell motility via receptor-mediated signaling pathways and is thought to be connected to tumor progression and metastasis. Using in vivo models, we showed that critical neovascularization responded to a biological amount of AMF. This angiogenic activity was fixed by specific inhibitors against AMF. AMF stimulated in vitro motility of human umbilical vein endothelial cells (HUVECs), inducing the expression of cell surface AMF receptor localizing a single predominant perinuclear pattern closely correlated with its motile ability. AMF also elicited the formation of tube-like structures mimicking angiogenesis when HUVECs were grown in three-dimensional type I collagen gels. We further immunohistochemically detected AMF receptors on the surrounding sites of newborn microvessels. These findings suggest that AMF is a possible tumor progressive angiogenic factor which may act in a paracrine manner for the endothelial cells in the clinical neoplasm, and it will be a new target for anti angiogenic treatment. PMID- 11414700 TI - Quantitative trait loci for body weight, blood pressure, blood glucose, and serum lipids: linkage analysis with wild rats (Rattus norvegicus). AB - To study polygenetically inherited human diseases like hypertension, inbred rat strains are usually the preferred models. Because many inbred generations under optimized environmental conditions may have led to the survival of "silent" disease genes, we used a cross between one wild rat and genetically hypertensive SHR rats to analyze quantitative trait loci (QTLs) of blood pressure and related traits. The (Wild x SHR)F1 hybrids were transferred into a pathogen-free environment by wet-hysterectomy and were backcrossed onto SHR to generate first backcross hybrids (BC1). Progeny from one F1 female (n = 72) were phenotypically and genetically characterized to map QTLs. Significant, subsignificant, and suggestive evidence was found for more sex-specific than common linkage of blood pressure and most blood-pressure-related traits. Male- and female-specific regions were determined on different chromosomes for blood pressures (Chrs. 2 and 7 vs 5 and 11), body weight (Chrs. 10 vs 18), and blood glucose (Chr. 17 vs 20). A linkage in both males and females was shown for serum triglycerides on chromosomes 6 and 17, respectively, and blood glucose on chromosome 15. For serum total cholesterol, a significant linkage was found on chromosome 14 only in males. Our findings not only indicate the complex character of quantitative traits per se but also show impressively their dependence on sex, age, and strains in cosegregation analysis. PMID- 11414701 TI - c-jun cooperates with SV40 T-antigen to sustain MMP-2 expression in immortalized cells. AB - The c-jun gene is a major regulator of proliferative and stress responses of both normal and transformed cells. In general, during immortalization/transformation c jun cooperates with oncogenic signals rather than acting as an oncogene itself. Here we report a novel example of this cooperation, the requirement for c-jun to sustain expression of the matrix metalloproteinase-2 (MMP-2) gene in cells immortalized by SV40 large T-antigen (TAg). MMP-2 encodes a type IV collagenase that is secreted by cells within normal and tumor microenvironments. We used wild type and c-jun null primary and TAg-immortalized mouse embryonic fibroblasts (mEFs) to investigate the importance of c-jun for the regulation of this activity, and observed that c-jun is essential for MMP-2 expression in immortalized but not primary mEFs. This finding directly demonstrates a cooperative interaction of c-jun with an oncogene, and suggests that TAg dependent immortalization/transformation may require other c-Jun/AP-1-dependent genes. PMID- 11414702 TI - Establishment of an expression cloning system for CD4+ T cell epitopes. AB - We previously reported an epitope presenting vector, pCI, a derivative of a human invariant chain (Ii) expression vector, in which the class II associated invariant chain peptide (CLIP, Ii p89-101) could be substituted with antigenic peptides. In the current study, we used this vector to develop a new expression cloning system to identify CD4+ T cell epitopes. We inserted double-stranded oligo DNAs of randomized sequences into this vector and prepared an epitope presenting library which loads randomized 13-mer peptides onto HLA class II molecules coexpressed in COS-7 cells. Utilizing this library, we isolated a cross reactive epitope recognized by a glutamic acid decarboxylase (GAD) 65 autoreactive T cell clone established from a patient with insulin-dependent diabetes mellitus. Although the newly identified epitope (PVQLSNQWHVVGATF) was far different from the original epitope, GAD65 p116-128 (NILLQYVVKSFDR), it did have the capacity to stimulate the T cell clone comparable to that of the original GAD epitope. Our system may be applicable not only for identifying of cross-reactive epitopes for CD4+ T cells of known specificity, but also for detection of epitopes stimulatory for CD4+ T cells the epitopes of which are unknown. PMID- 11414703 TI - The fission yeast ortholog of the coregulator SKIP interacts with the small subunit of U2AF. AB - The mode of action of transcriptional coregulators may involve the recruitment of spliceosome components. Using the two-hybrid screen, we examined the interaction partners of spSNW1, the S. pombe ortholog of the human coregulator SNW1/SKIP/NCoA 62, and found it to interact with the small subunit of the splicing factor U2AF (spU2AF23). The interaction involves the C-terminal parts of spU2AF23 and spSNW1. Tagged variants of both proteins were expressed in S. pombe and the interaction was proved by coprecipitation in nuclear extracts. This interaction would explain the finding of SKIP in nuclear speckles (Mintz, P. J., et al., EMBO J. 18, 4308 4320, 1999) and in reconstituted spliceosomes (Neubauer, G., et al., Nat. Genet. 20, 46-50, 1998). We deleted the spSNW1 gene in the diploid strain and demonstrated that spSNW1 is an essential gene in S. pombe. PMID- 11414704 TI - Cloning, characterization, and expression of human LIG1. AB - Growth factor receptors are frequently amplified and over-expressed in various human cancers. Recently, a Drosophila cell surface protein, Kekkon-1, was found to participate in an epidermal growth factor (EGF) driven negative feedback loop. Kekkon-1 is induced by EGF, binds to the EGF-receptor, and inhibits receptor mediated signaling. Here, we have searched for human genes with homologies to Kekkon-1 and identified human LIG1. The gene is the human homologue of mouse Lig 1 and is located on chromosome band 3p14, a region frequently deleted in various human cancers. It is predicted to encode a transmembrane cell-surface protein with extracellular leucine-rich repeats and immunoglobulin-like domains. LIG1 mRNA was detected in all tissues analyzed. The highest and lowest relative expression levels were found in brain and spleen, respectively, and differed by more than 200-fold. Taken together, our data are compatible with a role for LIG1 as a growth and tumor suppressor in human tissues. PMID- 11414705 TI - A protective role of PKCepsilon against TNF-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in glioma cells. AB - To elucidate the molecular mechanism(s) involved in the TRAIL-induced apoptosis sensitivity, we conducted the following experiments utilizing TRAIL-sensitive and -resistant glioma cells. We examined the expression of TRAIL receptors mRNA, but no significant differences were detected in those cells. TRAIL-resistant cells were sensitized to TRAIL-induced apoptosis by staurosporine pretreatment and preferentially expressed PKCepsilon. Since several lines of evidence suggest that PKC may play a protective role for apoptosis, we analyzed the involvement of PKCepsilon in TRAIL-induced apoptosis by an adenovirus vector expression system. We found that TRAIL susceptibility was augmented by the expression of a dominant negative PKCepsilon in TRAIL-resistant cells. Conversely, PKCepsilon introduction in TRAIL-sensitive cells resulted in the reduction of TRAIL-induced apoptosis. Taken together, these data suggest that PKCepsilon may be a regulator of susceptibility to TRAIL-induced apoptosis in gliomas and probably other malignancies. PMID- 11414706 TI - Molecular cloning and characterization of WNT3A and WNT14 clustered in human chromosome 1q42 region. AB - Human WNT3A and WNT14 cDNAs were cloned and characterized. WNT3A and WNT14 encoded WNT family protein of 352 and 365 amino acids, respectively. The 3.0-kb WNT3A mRNA was moderately expressed in placenta, and the 4.4-kb WNT14 mRNA was moderately expressed in skeletal muscle and heart. Although WNT3A mRNA was not detected in 35 human cancer cell lines, WNT14 mRNA was expressed in gastric cancer cell lines TMK1, MKN7, MKN45 and KATO-III. WNT3A and WNT14 genes, clustered in the head to head manner with an interval of about 58.0 kb, were mapped to human chromosome 1q42 region by fluorescence in situ hybridization. WNT3 and WNT15, clustered in human chromosome 17q21 region, are related genes of WNT3A and WNT14, respectively. WNT3A-WNT14 gene cluster and WNT3-WNT15 gene cluster might be generated due to duplication of ancestral gene cluster, just like WNT10A-WNT6 gene cluster and WNT10B-WNT1 gene cluster. Integration sites of mouse mammary tumor virus (MMTV) are located in the mouse chromosomal regions corresponding to these human WNT gene clusters. These results strongly suggest that unidentified nucleotide motif responsible for susceptibility to recombination might exist within the intergenic regions of these WNT gene clusters. PMID- 11414707 TI - TNFalpha induces and insulin inhibits caspase 3-dependent adipocyte apoptosis. AB - Regulation of fat cell number by apoptosis is proposed to be part of a normal physiological cycle in adipose growth and development. To investigate this process, cultured rat adipocytes were treated with various concentrations of tumor necrosis factor alpha (TNFalpha) and/or insulin to determine the roles of these factors in adipocyte apoptosis. The cells were analyzed by flow cytometry using a TUNEL assay. TNFalpha increased adipocyte apoptosis in a dose-dependent fashion. TNFalpha-mediated apoptosis was detectable within 6 h of treatment and continued to increase with time. Decreasing media insulin concentration from 8.5 to 0.85 nM resulted in increased adipocyte apoptosis, whereas high doses of insulin protected adipocytes from TNFalpha-induced apoptosis. TNFalpha-activated apoptosis was accompanied by an increase in caspase 3 activity and could be inhibited by a caspase 3-specific inhibitor. These data suggest that adipose tissue cell number is regulated, in part, by an apoptotic signaling pathway that involves TNFalpha, insulin, and caspase 3. PMID- 11414708 TI - Loss of calcitonin binding in rat is not related to calcitonin receptor gene abnormality. AB - C-cell tumors occur frequently (50%) in old WAG/Rij rats. Interestingly, genetically transmitted loss of CT binding sites in the kidney has also been demonstrated in WAG/Rij rats. To determine if these issues are resulted from mutation of calcitonin receptor (CTR), we analyzed the CTR genomic abnormality in WAG/Rij rat. We demonstrated that both Wistar and WAG/Rij rats expressed type-C1a CTR by RT-PCR analysis and their mRNA expressions were approximately equal by Northern blotting analysis. Direct sequence of RT-PCR products for CTR showed no different nucleotide sequences between the two strains. There were three polymorphisms at the first transmembrane domain and the fourth intracellular membranes, which are different from Sprague-Dawley rat. We concluded that the loss of CT binding in WAG/Rij rat is not related to CTR gene abnormality. Abnormal system of CTR amino acid modification may be occurred in WAG/Rij rat. PMID- 11414709 TI - FMRFamide-related neuropeptides are agonists of the orphan G-protein-coupled receptor GPR54. AB - We have isolated and determined the coding sequences of human and mouse orthologs of the rat orphan G-protein-coupled receptor GPR54. Mouse and rat GPR54 are nearly 95% identical to each other, and both are approximately 85% identical to human GPR54 at the amino acid level. Screening of agonists for GPR54 identified several invertebrate neuropeptides of the RFamide and RWamide family that were able to activate GPR54 at microM range through the G(alpha)q pathway. Substitution analysis showed that the C-terminal optimal sequence of GPR54 activating peptides is Gly-Leu-Arg-Trp-NH2. Northern analysis of human GPR54 detected expression in several peripheral tissues and many regions of the central nervous system. PMID- 11414710 TI - Cloning and characterization of the human stearoyl-CoA desaturase gene promoter: transcriptional activation by sterol regulatory element binding protein and repression by polyunsaturated fatty acids and cholesterol. AB - Stearoyl-CoA desaturase (SCD) is a microsomal enzyme required for the biosynthesis of oleate (C18:1) and palmitoleate (C16:1) which are the major monounsaturated fatty acids of membrane phospholipids, triglycerides and cholesterol esters. Previously the full-length human skin cDNA was sequenced and the exon and intron structure of the single functional SCD gene determined. Here we report on the cloning and characterization of the promoter region of the human SCD gene. The human promoter structure is very similar to that of the mouse SCD1 isoform and contains conserved regulatory sequences for the binding of several transcription factors including the sterol regulatory element binding protein (SREBP), CCAAT enhancer binding protein-alpha (C/EBPalpha) and nuclear factor-1 (NF-1) that have been shown to transactivate the transcription of the mouse SCD1 gene. Polyunsaturated fatty acids and cholesterol decreased the SCD promoter luciferase activity when transiently transfected into HepG2 cells. The decrease in promoter activity correlated with decreases in endogenous SCD mRNA and protein levels. Cotransfection experiment in HepG2 cells showed transactivation of the SCD promoter-luciferase activity by an expression vector containing SREBP-1a and 1c. Our studies indicate that the transcription of the human SCD gene is repressed by polyunsaturated fatty acids and cholesterol and that SREBP plays a role in the transcriptional activation of this gene. PMID- 11414711 TI - Beta2-adrenergic receptor/cyclic adenosine monophosphate (cAMP) leads to JNK activation through Rho family small GTPases. AB - Gi- and Gq-coupled G protein-coupled receptors (GPCRs) have been shown to activate c-Jun N-terminal kinase (JNK), a subfamily of mitogen-activated protein kinases (MAPKs), through Rho family small GTPases in mammalian cells. We investigated the signaling pathway linking the Gs-coupled beta2-adrenergic receptor with JNK, using smooth muscle DDT1 MF-2 cells, which natively express the beta2-adrenergic receptor. Stimulation of the beta2-adrenergic receptor activated JNK in a time-dependent manner, and a cell-permeable cyclic adenosine monophosphate analogue (8-Br-cAMP) activated JNK. The beta2-adrenergic receptor- or 8-Br-cAMP-induced activation of JNK required Rho family small GTPases. Also, the beta2-adrenergic receptor or 8-Br-cAMP induced activation of Rho family small GTPases. These results demonstrate that the beta2-adrenergic receptor/cAMP leads to JNK activation through Rho family small GTPases in DDT1 MF-2 cells. Activation of Rho family small GTPases may provide a common step in GPCR-mediated JNK activation. PMID- 11414712 TI - The brain basis of a "consciousness monitor": scientific and medical significance. AB - Surgical patients under anesthesia can wake up unpredictably and be exposed to intense, traumatic pain. Current medical techniques cannot maintain depth of anesthesia at a perfectly stable and safe level; the depth of unconsciousness may change from moment to moment. Without an effective consciousness monitor anesthesiologists may not be able to adjust dosages in time to protect patients from pain. An estimated 40,000 to 200,000 midoperative awakenings may occur in the United States annually. E. R. John and coauthors present the scientific basis of a practical "consciousness monitor" in two articles. One article is empirical and shows widespread and consistent electrical field changes across subjects and anesthetic agents as soon as consciousness is lost; these changes reverse when consciousness is regained afterward. These findings form the basis of a surgical consciousness monitor that recently received approval from the U.S. Food and Drug Administration. This may be the first practical application of research on the brain basis of consciousness. The other John article suggests theoretical explanations at three levels, a neurophysiological account of anesthesia, a neural dynamic account of conscious and unconscious states, and an integrative field theory. Of these, the neurophysiology is the best understood. Neural dynamics is evolving rapidly, with several alternative points of view. The field theory sketched here is the most novel and controversial. PMID- 11414713 TI - Invariant reversible QEEG effects of anesthetics. AB - Continuous recordings of brain electrical activity were obtained from a group of 176 patients throughout surgical procedures using general anesthesia. Artifact free data from the 19 electrodes of the International 10/20 System were subjected to quantitative analysis of the electroencephalogram (QEEG). Induction was variously accomplished with etomidate, propofol or thiopental. Anesthesia was maintained throughout the procedures by isoflurane, desflurane or sevoflurane (N = 68), total intravenous anesthesia using propofol (N = 49), or nitrous oxide plus narcotics (N = 59). A set of QEEG measures were found which reversibly displayed high heterogeneity of variance between four states as follows: (1) during induction; (2) just after loss of consciousness (LOC); (3) just before return of consciousness (ROC); (4) just after ROC. Homogeneity of variance across all agents within states was found. Topographic statistical probability images were compared between states. At LOC, power increased in all frequency bands in the power spectrum with the exception of a decrease in gamma activity, and there was a marked anteriorization of power. Additionally, a significant change occurred in hemispheric relationships, with prefrontal and frontal regions of each hemisphere becoming more closely coupled, and anterior and posterior regions on each hemisphere, as well as homologous regions between the two hemispheres, uncoupling. All of these changes reversed upon ROC. Variable resolution electromagnetic tomography (VARETA) was performed to localize salient features of power anteriorization in three dimensions. A common set of neuroanatomical regions appeared to be the locus of the most probable generators of the observed EEG changes. PMID- 11414714 TI - A field theory of consciousness. AB - This article summarizes a variety of current as well as previous research in support of a new theory of consciousness. Evidence has been steadily accumulating that information about a stimulus complex is distributed to many neuronal populations dispersed throughout the brain and is represented by the departure from randomness of the temporal pattern of neural discharges within these large ensembles. Zero phase lag synchronization occurs between discharges of neurons in different brain regions and is enhanced by presentation of stimuli. This evidence further suggests that spatiotemporal patterns of coherence, which have been identified by spatial principal component analysis, may encode a multidimensional representation of a present or past event. How such distributed information is integrated into a holistic precept constitutes the binding problem. How a precept defined by a spatial distribution of nonrandomness can be subjectively experienced constitutes the problem of consciousness. Explanations based on a discrete connectionistic network cannot be reconciled with the relevant facts. Evidence is presented herein of invariant features of brain electrical activity found to change reversibly with loss and return of consciousness in a study of 176 patients anesthetized during surgical procedures. A review of relevant research areas, as well as the anesthesia data, leads to a postulation that consciousness is a property of quantum-like processes, within a brain field resonating within a core of structures, which may be the neural substrate of consciousness. This core includes regions of the prefrontal cortex, the frontal cortex, the pre- and paracentral cortex, thalamus, limbic system, and basal ganglia. PMID- 11414715 TI - Commentary on E. R. John et al. "Invariant reversible QEEG effects of anesthetics" and E. R. John "A field theory of consciousness" . PMID- 11414716 TI - Anesthesia: the "other side" of consciousness. PMID- 11414717 TI - Anesthesia--a descent or a jump into the depths? PMID- 11414718 TI - The power of observation. PMID- 11414719 TI - Identifying the neural correlates of consciousness: strategies with general anesthetics. PMID- 11414720 TI - Commentary on E. R. John et al. "Invariant reversible QEEG effects of anesthetics" and E. R. John "A field theory of consciousness". PMID- 11414722 TI - Testing Stereotype Threat: Does Anxiety Explain Race and Sex Differences in Achievement? AB - Steele's (1992, 1997) stereotype-threat theory attempts to explain underperformance of minority students in academic domains and of women in mathematics. Steele argues that situational self-relevance of negative group stereotypes in testing situations increases the anxiety these students experience and that these differential anxiety levels explain performance differences. Research shows that manipulation of stereotype threat can affect academic performance. However, there has been little research testing whether anxiety does at least partially explain the relationship between race and achievement. The goal of this study was to examine whether anxiety will explain racial differences in academic performance and gender differences in math performance in the context of a nationally representative sample of high school seniors. Partial mediation was observed, with anxiety explaining significant portions of the racial differences in academic performance. Anxiety also partially explained sex differences in math achievement, although the effect sizes were very small. These results provide general support for Steele's stereotype-threat hypothesis. Copyright 2001 Academic Press. PMID- 11414723 TI - Student Goal Orientation and Interest: Effects on Students' Use of Self-Regulated Learning Strategies. AB - This study examined how goal orientation and interest together affected students' use of learning strategies. Ninety-three Grade 11 students participated in this study. This study was a 2 x 2 between-groups factorial using a post-test-only control group design. There were two independent variables, Goal orientation and Interest. There were three dependent variables: Total Main Ideas, Use of Other Cognitive Strategies, and Metacognition. There were significant main effects for Interest and Goal Orientation on the main-idea selection measure: high-interest students selected more main ideas than low-interest students, and rewarded students outperformed learning goal-oriented students. There was a main effect for Interest on Metacognition: high-interest students reported using more metacognitive strategies than low-interest students. Interpretation of these results is discussed from a theoretical perspective and practical applications are offered as well. Copyright 2001 Academic Press. PMID- 11414724 TI - Gender Differences in Arithmetic Strategy Use: A Function of Skill and Preference. AB - This study was designed to examine whether first-grade boys' use of retrieval and first-grade girls' use of manipulatives reflected gender differences in their abilities to use these strategies or gender differences in preferences for strategy use. Eighty-four first-grade students, 42 boys and 42 girls, from two suburban elementary schools participated in this study. The children solved basic arithmetic problems under two conditions: a free-choice condition in which they were allowed to solve the problems any way they preferred and a game condition in which the children's strategy use was constrained so that all children used the same strategies on the same arithmetic problems. Strategy use during the free choice session replicated the findings of earlier research indicating that girls tend to use strategies utilizing manipulatives and boys tend to use retrieval. During the game condition, when we controlled the types of strategies children used on different problems we found that boys were as able as girls to calculate solutions using manipulatives. Girls, however, were not as capable as boys in their retrieval of answers to arithmetic problems from memory. No differences were found in error rates or speed of retrieval. Gender differences were found in the variability of correct retrieval, with boys being significantly more variable than girls. Copyright 2001 Academic Press. PMID- 11414725 TI - Conceptions of Ability and Achievement Goals in Physical Education: Comparisons of American and Chinese Students. AB - This study focused on comparisons of American and Chinese students in conceptions of ability and achievement goals in physical education. Three hundred eight American students and 371 Chinese students in 4th, 8th, and 11th grades completed questionnaires assessing their conceptions of ability and achievement goals. Results indicated that for both American and Chinese students, as they progressed through the grades they tended to develop a differentiated conception of ability and to become more ego-oriented. Achievement goals were related to conceptions of ability. Cultural differences were found in the two variables. The findings offer additional empirical support to the view that goal theory is relevant to physical education settings and that achievement-related cognitions vary as a function of children's age and cultural background. Copyright 2001 Academic Press. PMID- 11414726 TI - Gender Differences in Writing Motivation and Achievement of Middle School Students: A Function of Gender Orientation? AB - The aim of this study was to determine whether gender differences in the writing motivation and achievement of middle school students (N = 497) are a function of gender-stereotypic beliefs rather than of gender. Girls reported stronger writing self-efficacy, writing self-concept, self-efficacy for self-regulation, value of writing, and task goals, and they received higher grades in language arts. Boys reported stronger performance-approach goals. All gender differences favoring girls in writing motivation and achievement were rendered nonsignificant when feminine orientation beliefs were controlled. Findings suggest that a feminine orientation is adaptive in the area of writing, whereas a masculine orientation is beneficial when escorted by a feminine orientation. Results are interpreted from the perspective of A. Bandura's (1986) social cognitive theory. Copyright 2001 Academic Press. PMID- 11414727 TI - Student Interest, Grading Leniency, and Teacher Ratings: A Conceptual Analysis. AB - This study explored the extent to which student interest and grading leniency were predictive of teacher ratings. Unlike other teacher rating research, however, this study measured student interest at the beginning and end of seven sections of two courses; further, grading leniency was explicitly defined and measured, not inferred from expected grade and workload. Data indicate that precourse interest was positively associated with expected grade, but was not predictive of ratings, nor did it moderate the expected grade-rating association. Rather, interest change was positively associated with expected grade and predictive of ratings. Further, interest change and grading leniency provided incremental variance in ratings, beyond that provided by expected grade. Copyright 2001 Academic Press. PMID- 11414728 TI - Semantic Domains of Rational Numbers and the Acquisition of Fraction Equivalence. AB - This study examined how the semantic meanings of rational numbers embodied in graphical representations pose a constraint on children's construction of the concept of fraction equivalence. A test consisting of graphical representations representing the part-whole and measuring semantic meaning was given to 205 fifth graders and 208 sixth-graders from China. Results showed that the fifth-graders' overall performance on equivalent fraction items was poor across the semantic domains of rational numbers. The sixth-graders showed significantly improved performance on the equivalent fraction items representing the part-whole relation but not on those representing the measure aspect of rational number. The results provide evidence that semantic meanings of fractional numbers in different relevant contexts also constitute a source of difficulty in children's constructing the concept of fraction equivalence, in addition to the multiplicative nature of the concept. Copyright 2001 Academic Press. PMID- 11414729 TI - Effects of Headings on Text Processing Strategies. AB - This experiment addressed the question of how headings influence readers' memories for text content. College students read and recalled a 12-topic expository text. Half of the participants were trained to construct a mental outline of the text's topic structure as they read and then use their mental outlines to guide their recall attempts. The remaining participants did not receive such training. Half of the participants read a text containing headings before every subsection; the other half read the same text without headings. The results were that participants who received training and/or read the text with headings remembered text topics and their organization better than participants who received no training and read the text without headings. The results support the hypothesis that signals induce a change in readers' strategies for encoding and recalling text. Copyright 2001 Academic Press. PMID- 11414731 TI - Mice disrupted for the KvLQT1 potassium channel regulator IsK gene accumulate mature T cells. AB - The IsK protein associates with KvLQT1 potassium channels to generate the slow component of the outward rectifying K(+) current involved in human cardiac repolarization. Mutations in either KCNE1 (encoding IsK) or KCNQ1 (encoding KvLQT1) genes have been associated with the long QT syndrome, a genetic disorder leading to prolonged cardiac repolarization and sudden death. We now report that the IsK protein is also involved in mature T cell homeostasis. In KCNE1 gene knockout mice, we observed a significant increase in the T cell compartment. Thymus and peripheral lymphoid organs of KCNE1-/- mice displayed a significant increase in mature T cells. The immunological phenotype of KCNE1-/- is age dependent and only expressed in adult mice. Both IsK and KvLQT1 mRNA are expressed in murine thymus. Our data suggest that, in addition to its role in myocardial repolarization, the IsK-KvLQT1 tandem also plays a crucial role in T cell homeostasis. PMID- 11414732 TI - Idiotypic-anti-idiotypic B cell interactions generated against a protective antigen of a morbillivirus in mice. AB - The idiotypic network theory (N. K. Jerne, Ann. Immunol. 125, 373-389, 1974) predicts that any antibody that can be made by an individual would have its preexisting specific complementary B cells in its germline repertoire. We transplanted syngeneic BALB/c mice with live hybridoma cells and demonstrated the simultaneous presence of interacting idiotypic and anti-idiotypic B cells in an individual animal by immuno-cytoadherence assays. Furthermore, we demonstrate that interacting B cells displaying idiotypic and anti-idiotypic antibodies are subjected to lysis by complement. It is therefore tempting to speculate that this complement-sensitive interaction between idiotypic and complementary anti idiotypic B cells in vivo may provide a mechanism for the regulation of B cell populations. PMID- 11414733 TI - Dendritic cells issued in vitro from bone marrow produce PGE(2) that contributes to the immunomodulation induced by antigen-presenting cells. AB - Given that preliminary work has indicated that prostaglandins can play a role in modulating dendritic cell (DC) functions, we addressed the prostaglandin E(2) (PGE(2)) biosynthetic capacity of mouse DC produced in vitro from bone marrow cells. We observed production of significant amounts of PGE(2), which was reduced by at least 80% when cells were incubated in the presence of indomethacin, a COX 1 preferential inhibitor. Indeed, when tested by Western blot analysis with specific COX-1 and COX-2 antibodies, only COX-1 expression could be detected in the bone marrow (BM)-DC. For lipopolysaccharide (LPS)-treated BM-DC, inhibition of PGE(2) production by indomethacin or by NS-398 (a COX-2-selective inhibitor) used alone was less potent. After LPS treatment of BM-DC, COX-1 and COX-2 expression was potent, and inhibition of PGE(2) synthesis needed the presence of both indomethacin and NS-398. We also observed that exogenous PGE(2) diminished the expression of MHC class II molecules by BM-DC and that prostaglandin and indomethacin had antagonistic effects on cell proliferation during the mixed lymphocyte reaction using BM-DC as stimulatory cells. This assessment of PGE(2) suggests that endogenous PGE(2) produced by DC might play a role as an immunomodulating factor during the immune response. This hypothesis is sustained by the fact that IL-12 production by BM-DC is modulated by exogenous PGE(2) as well as endogenous prostaglandin, since either the addition of exogenous PGE(2) or the presence of LPS (which increases endogenous PGE(2) synthesis) decreases IL 12 production, while NS-398 (which decreases LPS-induced PGE(2) synthesis) increases IL-12 synthesis. PMID- 11414734 TI - A "chimeric" C57l-derived Ly49 inhibitory receptor resembling the Ly49D activation receptor. AB - Ly49D is a natural killer (NK) cell activation receptor that is responsible for differential mouse inbred strain-determined lysis of Chinese hamster ovary (CHO) cells. Whereas C57BL/6 NK cells kill CHO, BALB/c-derived NK cells cannot kill because they lack expression of Ly49D. Furthermore, the expression of Ly49D, as detected by monoclonal antibody 4E4, correlates well with CHO lysis by NK cells from different inbred strains. However, one discordant mouse strain was identified; C57L NK cells express the mAb 4E4 epitope but fail to lyse CHO cells. Herein we describe a Ly49 molecule isolated from C57L mice that is recognized by mAb 4E4 (anti-Ly49D). Interestingly, this molecule shares extensive similarity to Ly49D(B6) in its extracellular domain, but its cytoplasmic and transmembrane domains are identical to the inhibitory receptor Ly49A(B6), including a cytoplasmic ITIM. This molecule bears substantial overall homology to the previously cloned Ly49O molecule from 129 mice the serologic reactivity and function of which were undefined. Cytotoxicity experiments revealed that 4E4(+) LAK cells from C57L mice failed to lyse CHO cells and inhibited NK cell function in redirected inhibition assays. MHC class I tetramer staining revealed that the Ly49O(C57L)-bound H-2D(d) and lysis by 4E4(+) C57L LAK cells is inhibited by target H-2D(d). The structural basis for ligand binding was also examined in the context of the recent crystallization of a Ly49A-H-2D(d) complex. Therefore, this apparently "chimeric" Ly49 molecule serologically resembles an NK cell activation receptor but functions as an inhibitory receptor. PMID- 11414735 TI - Molecular cloning and expression pattern of porcine myeloid DAP12-associating lectin-1. AB - DAP12 is an ITAM-bearing membrane protein that is associated with activating receptors in natural killer cells, granulocytes, macrophages, and monocytes. Myeloid DAP12-associating lectin-1 (MDL-1) is a type II membrane protein that associates with DAP12. In this study, we report the molecular cloning of two isoforms of porcine MDL-1 cDNA from pulmonary alveolar macrophages. The porcine MDL-1 short form has 165 amino acids and 70% sequence identity with the mouse MDL 1 short form. The long form has 20 more amino acids in the stalk region and 71% sequence identity with human MDL-1 and 67% with the mouse MDL-1 long form. Porcine MDL-1 contains a conserved lysine in the transmembrane domain. There are six putative N-linked glycosylation sites in the MDL-1 long form. MDL-1 transcripts were detected exclusively in macrophages and monocytes by RT-PCR. When transfected into 293 cells, porcine MDL-1 is expressed on the cell surface associated with DAP12. PMID- 11414736 TI - Positive and negative consequences of soluble Fas ligand produced by an antigen specific CD4(+) T cell response in human carcinoma immune interactions. AB - The influence of a human CD4(+) T cell response in anti-carcinoma immune reactions remains largely uncharacterized. Here, we made use of a major histocompatibility complex (MHC) class-II-restricted, anti-ras oncogene-specific CD4(+) T cell line produced previously in vivo from a patient with metastatic carcinoma in a peptide-based phase I trial. Using this patient-derived T cell line as a potentially relevant cell type, we examined the consequences of the anti-carcinoma CD4(+) T cell response, with emphasis on specific lymphokines potentially important for the regulation of Fas/Fas ligand (FasL) interactions. Antigen (Ag)-specific CD4(+) T cells produced substantial amounts of IFN-gamma following recognition of MHC class-II-matched Ag-presenting cells expressing the cognate peptide. The IFN-gamma promoted significant upregulation of Fas on the surface of colon carcinoma cells and sensitized these targets to Fas-mediated apoptosis and Ag-specific CD8(+) cytotoxic T lymphocyte (CTL)-mediated lysis involving a Fas-based effector mechanism. Moreover, Ag-stimulated CD4(+) T cells secreted soluble FasL (sFasL), which induced the death of TNF resistant/refractory colon, breast, and ovarian carcinoma cells. Interestingly, although CD4(+)-derived sFasL expressed cytotoxic activity, the recovery of carcinoma cells which resisted Fas-mediated lysis displayed enhanced metastatic ability in vivo, compared with the unselected parental population, in an athymic mouse model. Thus, a tumor-specific CD4(+) T cell response may have both positive and negative consequences in human carcinoma via the production of proinflammatory cytokines such as IFN-gamma and/or sFasL that may (1) improve or facilitate CTL-target engagement, contact-independent effector mechanisms, and the overall lytic outcome and (2) potentially select for Fas-resistant tumor cells that escape immune destruction, which may thus impact the metastatic process. PMID- 11414737 TI - OX40 ligation enhances cell cycle turnover of Ag-activated CD4 T cells in vivo. AB - OX40 costimulates T cells, increases activated T cell longevity, and promotes memory acquisition. T cells activated in vivo with agonist anti-OX40 and ovalbumin have a unique pattern of survival and cell division compared to control cells, but are able to respond to recall Ag equally well. BrdU incorporation shows that early cellular division rates of the anti-OX40-treated and the control groups are similar. Nevertheless, more BrdU(+) Ag-specific T cells accumulate in lymphoid tissue upon anti-OX40 administration. Thus, OX40 ligation does not necessarily lead to increased cell cycle entry, but promotes the accumulation of dividing cells. However, CFSE staining shows that OX40 ligation allows cells to progress through more cellular division cycles, while control cells stall or die. Moreover, OX40 ligation leads to a proportional decrease in apoptotic Ag-specific T cells. Thus, OX40 ligation boosts immunity by promoting an increase in the number cell cycles completed, thereby increasing the life span of Ag-activated CD4 T cells. PMID- 11414738 TI - Adoptive transfer of a superantigen-induced hole in the repertoire of natural IgM secreting cells. AB - We have recently evaluated the host response to the bacterial toxin, protein A from Staphylococcus aureus (SpA), which has the capacity to interact with B-cell antigen receptors encoded by V(H) clan III genes via a conserved variable region framework surface. In these studies, intraperitoneal instillation of SpA induced a persistent T-cell independent loss of a large supraclonal set of susceptible lymphocytes, which includes clan III/V(H) S107 family-expressing B-1 cells and their antibody products. To determine whether these long-term effects could represent the influence of residual in vivo deposited superantigen, we have now performed adoptive transfer of peritoneal B cells from superantigen- and control treated donors. These studies demonstrated that mice that received cells from SpA treated donors also exhibited the same induced supraclonal hole in the expressed repertoire of natural IgM-secreting cells due to supraclonal deletion. These studies clarify the cellular mechanisms responsible for B-cell superantigen induced modification of the repertoires of in vivo polyclonal B-cell populations. PMID- 11414739 TI - XLP: one gene, many players. PMID- 11414740 TI - Chronic beryllium disease: T cell recognition of a metal presented by HLA-DP. PMID- 11414741 TI - Distinct interactions of the X-linked lymphoproliferative syndrome gene product SAP with cytoplasmic domains of members of the CD2 receptor family. AB - X-linked lymphoproliferative syndrome (XLP; Duncan's disease) is a primary immunodeficiency disease that manifests as an inability to regulate the immune response to Epstein-Barr virus (EBV) infection. Here we examine the ability of the product of the gene defective in XLP, SAP (DSHP/SH2D1A), to associate with the cytoplasmic domains of several members of the CD2 subfamily of cell surface receptors, including SLAM, 2B4, and CD84. While recruitment of SAP to SLAM occurred in a phosphorylation-independent manner, SAP was found to bind preferentially to tyrosine-phosphorylated cytoplasmic domains within 2B4 and CD84. Missense or nonsense mutations in the SAP open reading frame were identified in five of seven clinically diagnosed XLP patients from different kindreds. Four of these variants retained the ability to bind to the cytoplasmic tails of SLAM and CD84. While ectopic expression of wild-type SAP was observed to block the binding of SHP-2 to SLAM, mutant SAP derivatives that retained the ability to bind SLAM did not inhibit recruitment of SHP-2 to SLAM. In contrast, SAP binding to CD84 had no effect on the ability of CD84 to recruit SHP-2, but instead displaced SHP-1 from the cytoplasmic tail of CD84. These results suggest that mutations in the gene encoding the XLP protein SAP lead to functional defects in the protein that include receptor binding and SHP-1 and SHP-2 displacement and that SAP utilizes different mechanisms to regulate signaling through the CD2 family of receptors. PMID- 11414742 TI - Azodicarbonamide as a new T cell immunosuppressant: synergy with cyclosporin A. AB - We recently demonstrated that azodicarbonamide is an immunosuppressive compound that inhibits calcium mobilization in T lymphocytes. In this study, we show that azodicarbonamide prevents the progression of human CD4+ T lymphocytes into the G1 phase of the cell cycle, inhibits their blastogenesis, down-regulates their membrane expression of CD25 and CD69, and decreases their transcription of cytokine genes. Addition of the calcium ionophore A23187 completely restores T cell proliferation in the presence of azodicarbonamide. Furthermore, azodicarbonamide synergizes with cyclosporin A to inhibit CD4+ T cell proliferation. In conclusion, the immunosuppressive action of azodicarbonamide is mainly related to its effect on the calcium mobilization machinery and is synergistic with that of calcineurin inhibitors. PMID- 11414743 TI - T cell receptor Vbeta expression in patients with allergic asthma before and after repeated low-dose allergen inhalation. AB - The objective of this study was to identify disease-associated T cell subsets by characterizing the lung and blood T cell receptor (TCR) repertoires in allergic asthmatics before and after repeated low-dose allergen challenge. Peripheral blood lymphocyte (PBL) and bronchoalveolar lavage (BAL) samples were obtained from eight patients with allergic asthma before and after a period of repeated low-dose allergen inhalations. RT-PCR followed by Southern blot allowed the quantification of relative Vbeta gene segment usage. Thirteen healthy individuals served as controls at PBL level. PBL as well as BAL T cells of asthmatics displayed a higher usage of Vbeta3, Vbeta5.2, and Vbeta6.1-3 and a lower usage of Vbeta16, Vbeta18, and Vbeta19 compared to PBL of healthy controls. Interestingly, TCR Vbeta7 and Vbeta9 usage was significantly higher in BAL than in PBL in asthmatics before as well as after challenge. TCR repertoire alterations after allergen challenge differed between individuals, with relatively mild changes. PMID- 11414744 TI - Radioimmunotherapy with iodine-131 tositumomab in patients with low-grade non Hodgkin's B-cell lymphoma does not induce loss of acquired humoral immunity against common antigens. AB - Thirty-one previously untreated patients with follicular low-grade B-cell non Hodgkin's lymphoma expressing the CD20 antigen were treated with iodine-131 tositumomab therapy between 1996 and 1998. The therapy led to a temporary depletion of peripheral blood B-lymphocytes. Recovery of B-cells occurred in most cases by 3 to 6 months and in all patients by 12 months posttherapy. A temporary decline in T-cell subpopulations, but no reduction in serum immunoglobulin levels, could be observed. ELISA techniques were used to detect specific antibodies against rubella, mumps, varicella zoster, measles, and tetanus. Almost all patients remained seropositive against the different antigens during the 1- to 2-year follow-up. No significant reduction in antibody concentrations to tetanus or measles could be detected. The data show that acquired humoral immunity against common antigens appears to be preserved despite a temporary loss of B-lymphocytes. PMID- 11414745 TI - Cytokine and chemokine dysregulation in hyper-IgE syndrome. AB - Hyper-IgE syndrome is characterized by severe recurrent staphylococcal infections, eczema, bone abnormalities, and markedly elevated levels of immunoglobulin E (IgE). The genetic basis is not known and the central immunologic defect is largely undefined. Reduced neutrophil chemotaxis is often described, and variable T cell defects have been demonstrated in some patients. It has been hypothesized that hyper-IgE is associated with a Th1/Th2 imbalance. We wished to characterize cytokine and chemokine imbalances that might reflect the underlying disease process or reflect ongoing pathologic processes. Nine patients with hyper-IgE syndrome and six controls were studied. Radioimmunoassays, flow cytometry, and gene array analyses were performed to characterize cytokine and chemokine production. Hyper-IgE patients express more IL-12, while ENA-78, MCP-3, and eotaxin are markedly underexpressed. Underexpression of a set of chemokines could explain a number of features of hyper-IgE syndrome and may offer a new paradigm for the understanding of this disorder. PMID- 11414746 TI - Autoimmune-mediated vasculopathy. AB - The use of the immunosuppressive drug cyclosporine A (CsA) in solid organ transplantation can be associated with the development of vasculopathy as part of the complex immune response involved in chronic rejection, including autoimmune recognition. Although CsA can directly affect endothelial cells, this drug alters the T cell repertoire promoting autoimmune recognition. The present studies evaluated the ability of CsA-induced autoreactive T cells to mediate vascular lesions in syngeneic heart grafts. Graft vasculopathy developed in syngeneic heart grafts following either the primary induction of autoimmunity with CsA or the adoptive transfer of CsA-induced autoreactive T cells. Initially, an inflammatory response occurred in the medial wall of the small arterial vessels, accompanied by a perivascular lymphocytic infiltrate (including a lymphocytic infiltrate into the myocardium), followed by progression of vascular disease with endothelial cell proliferation. The development and progression of vascular disease correlated with the cytokine profile of the infiltrating lymphocytes with type 1 cytokines detected early and type 2 cytokines detected as the disease progressed. Initiation of this response correlated with upregulation of the target antigen recognized by the CsA-induced autoreactive T cells, the MHC class II-invariant chain peptide complex. This antigen complex, when upregulated on endothelial cells by interferon, allowed effective targeting by the autoreactive T lymphocytes. Strategies to inhibit the upregulation of MHC class II antigens by treatment of the recipients with chloroquine truncated the disease process. The results of these studies suggest that CsA-induced autoreactive mechanisms can contribute to the development of graft vasculopathy. PMID- 11414747 TI - The role of CD40-CD40 ligand (CD154) interactions in immunoglobulin light chain repertoire generation and somatic mutation. AB - To determine whether CD40 ligation influences the molecular and selective mechanisms that govern the development of the human Ig light chain repertoire, analysis of the Vkappa and Vlambda repertoires of CD19+ B cells obtained from a patient with X-linked hyper IgM syndrome (XHIM) and a nonfunctional CD154 was carried out. The nonproductive Vkappa and Vlambda repertoires were largely comparable to that of the normals with respect to V gene and J segment distribution as well as CDR3 length and VLJL joint complexity. Comparison of the nonproductive and productive repertoires indicated that a limited number of VL genes were positively and negatively selected in the XHIM patient. Although mutations were observed in the XHIM VL repertoires, the frequency of mutations was significantly lower than in normals. Typical targeting of these mutations into RGYW/WRCY motifs was significantly reduced and subsequent selection of RGYW/WRCY mutations, which is normally observed, was not found. These results indicate that CD40 ligation is not required for generation of the light chain repertoire, positive selection of some Vk rearrangements, negative selection of specific VL genes, and some degree of somatic mutation. Importantly, however, targeting of mutations to RGYW/WRCY motifs and subsequent selection of these mutated motifs does not occur in the absence of CD40 ligation. PMID- 11414748 TI - Restored antioxidant capacity parallels the immunologic and virologic improvement in children with perinatal human immunodeficiency virus infection receiving highly active antiretroviral therapy. AB - CD3+CD4+ T-lymphocyte numbers, viral load, and serum antioxidant capacity were evaluated in 20 children with perinatal human immunodeficiency virus (HIV) infection one month (T = -1) and one day (T = 0) before and one month (T = 1) and two months (T = 2) after a treatment switch to highly active antiretroviral therapy (HAART). Antioxidant capacity micromol/L) was evaluated by measuring the cuprous ion deriving from a known amount of cupric ion. Compared to control values (998 +/- 113 micromol/L), values in HIV-infected children were lower before HAART (T = -1, 848 +/- 211 micromol/L, P = 0.008; T = 0, 732 +/- 131 micromol/L, P < 0.0001), but similar during HAART (T = 1, 914 +/- 121 micromol/L, P = 0.089; T = 2; 957 +/- 155 micromol/L, P = 0.528; T = 1 and T = 2 vs T = 0, P < 0.0001). Immunologic and virologic improvement paralleled the restored antioxidant capacity. HAART may restore antioxidant capacity suppressing HIV, which inhibits antioxidant capacity. A positive feedback may be triggered since restored antioxidant capacity counterbalances the oxidative stress, which enhances lymphocyte apoptosis and HIV replication. PMID- 11414749 TI - Activation of peripheral blood neutrophils from patients with active advanced tuberculosis. AB - Activation of peripheral blood neutrophils (PMN) was investigated in order to determine whether they might contribute to the inflammatory process during active advanced tuberculosis. Receptors for the Fc portion of IgG (FcgammaR) (FcgammaRI, FcgammaRII, and FcgammaRIIIB), CD66 (degranulation marker), and receptors for tumor necrosis factor-alpha (TNF-R55 and TNF-R75) were analyzed on PMN obtained from normal controls and tuberculosis patients (TB-PMN). Functional parameters such as cytotoxicity, superoxide anion generation triggered by N-formyl-methionyl leucyl-phenyl-alanine (FMLP), and TNF-alpha and IL-1beta production were evaluated. A high expression of TNF-R55, CD66, and FcgammaRIIIB and the appearance of FcgammaRI were detected in TB-PMN. In addition, cytotoxicity, superoxide anion release, and TNF-alpha and IL-1beta production were enhanced in TB-PMN. Thus, in tuberculosis, the activation of PMN outside the focus of infection strongly suggests the possibility of a systemic inflammation that could modulate the inflammatory response. PMID- 11414750 TI - Mechanisms for induction of immunosuppression during experimental cryptococcosis: role of glucuronoxylomannan. AB - In previous work we have demonstrated that spleen mononuclear (Spm) cells from rats obtained 14 days after infection with Cryptococcus neoformans showed a diminution in proliferative response to Concanavalin A (Con A). In this study we further investigate some characteristics of the Spm cell population involved in the immunosuppressor phenomenon induced by C. neoformans. We observed that unstimulated Spm cells expressing T-cell receptor (TCR+) from infected rats were reduced in number after 96 h of culture. When the Spm cells from infected rats were stimulated with Con A, increased production of IL-10, reduced levels of IL 2, and decreased CD11a surface expression were shown. These immunosuppressor phenomena were also observed when the capsular polysaccharide, glucuronoxylomannan (GXM), was added to cultures of Spm cells from normal rats. However, GXM had a more pronounced effect in reducing the number of cells surviving in culture than that observed during infection and produced an increase in IL-4 production by Con-A-stimulated Spm cells. Addition of anti-IL-10 monoclonal antibody to cultures restored the lymphoproliferation of Spm cells from infected animals, indicating that IL-10 production is a suppressor mechanism of cell-mediated immunity during experimental infection. The results presented here indicate that at least two mechanisms mediate the nonspecific suppression in this model of cryptococcosis: IL-10 production and diminution of the number of T cells. GXM could be involved, since it has a pronounced effect in the reduction of Spm cells in vitro. PMID- 11414751 TI - Increased CCR5 and CXCR4 expression in Ethiopians living in Israel: environmental and constitutive factors. AB - HIV coreceptors play a major role in determining susceptibility and HIV cell tropism. The present work studied whether the high expression of these coreceptors found on lymphocytes and monocytes of Ethiopian immigrants to Israel (ETH) is the result of environmental and/or constitutive genetic factors. The study of 26 ETH shortly after their arrival to Israel (new ETH), 22 ETH in Israel over 7 years (old ETH), and 20 Caucasian Israelis (non-ETH) enabled us to address this issue. The new ETH had elevated levels of activated HLA-DR+CD4+ and CD38+CD8+ cells in comparison with both old ETH and non-ETH groups (P < 0.01), most probably related to chronic helminthic infections. Surface CCR5 expression, i.e., the percentage of CCR5+ cells and the number of CCR5 molecules/cell, was higher (2- to 3- and 8- to 31-fold, respectively) in activated than in nonactivated CD4+ cells, in all groups. However, CCR5 expression, in both activated and nonactivated CD4+ cells, was higher in both ETH groups than in the non-ETH group. CXCR4 expression was higher in nonactivated CD4+ cells in all groups and was also higher in both ETH groups, in both activated and nonactivated CD4+ cells, than in the non-ETH group. These findings suggest that constitutive factors, in addition to immune activation caused by environmental factors, account for the elevated expression of CCR5 and CXCR4 on CD4+ cells of ETH. This increased HIV coreceptor expression may make ETH more susceptible to HIV infection and may account in part for the rapid spread of AIDS in Ethiopia and the rest of Africa as well. PMID- 11414752 TI - Complement-regulator factor H and related proteins in otitis media with effusion. AB - Otitis media with effusion (OME) is a common disease in childhood. It is characterized by chronic inflammation in which the proinflammatory activity of the complement (C) system is one of the underlying factors. The C system becomes strongly activated in the middle ear effusion (MEE) fluid, but the reasons for this are not known. Here we demonstrate by using complement Bb fragment ELISA that MEE specimens strongly activate the alternative C pathway (AP) in normal human serum (NHS). Some of the MEEs were also found to promote lysis of rabbit erythrocytes by NHS. These findings indicated a disturbance in the fluid-phase regulation of the AP in MEE. The main regulator of the AP, factor H (FH), and proteins structurally related to it (FHL-1, FHR-1, -2, -3, and -4) were present in the MEE fluids of OME patients. Relative to serum, the FHR proteins were more abundant in the MEEs. In addition, we detected the recently discovered 65-kDa FH related protein FHR-5 in the MEE. The FHR proteins share binding sites with FH in the C3d region of C3b. Thus they may compete with FH in binding to C3b and interfere with the regulatory activity of FH. Consequently, a disturbance in AP control in the MEE may lead to an ongoing excessive C activation and inflammation in OME. PMID- 11414753 TI - Three loci modify growth of a transgene-induced mammary tumor: suppression of proliferation associated with decreased microvessel density. AB - In earlier studies it was observed that the genetic background significantly affected the phenotype of a transgene-induced mammary tumor. Tumors arising in an (I/LnJ x PyMT) F1 hybrid background appeared earlier than in the FVB/N-TgN(MMTV PyVT)(634Mul) parent, but accumulated less tumor mass, indicating a net decrease in tumor growth. Quantitative genetic mapping in a backcross identified three loci that were associated with the decreased proliferative capacity of the I/LnJ F1 tumors. Molecular analysis of the tumors suggests that these loci may act by restricting the tumor's ability to recruit microvessels. The three loci, designated Mmtg1-3, are unlinked to the angiogenic genes Fgf2, Flt1, Flk4, Flk1, Vegf, and Vegfc, as well as the precursors of the endogenous antiangiogenic molecules angiostatin and endostatin. The Mmtg loci may therefore provide novel targets for antiangiogenic therapeutic strategies. PMID- 11414754 TI - A novel human amino acid transporter, hNAT3: cDNA cloning, chromosomal mapping, genomic structure, expression, and functional characterization. AB - Amino acid transporters are proteins that transport amino acids across the membrane. We report here the isolation and characterization of a novel human cDNA clone encoding a protein of 547 amino acids. This protein shares approximately 50% amino acid sequence homology with the amino acid transporters mouse mNAT and its orthologs, rat SN1 and human g17, and mouse GlnT/ATA1 and ATA2. Expression of this cRNA in Xenopus oocytes revealed that the strongest transport activities were specific for l-alanine. In addition, hNAT3 is a Na(+)- and pH-dependent, low affinity transporter and partially tolerates substitution of Na(+) by Li(+). Since this protein has sequence and functional similarities to the previously identified system N amino acid transporters, we named this protein hNAT3. The genomic DNA sequence encoding the transcript of hNAT3 spans over 14 kb with 16 exons and 15 introns. Using fluorescence in situ hybridization, we mapped the hNAT3 gene to human chromosome 12q12-q13. By RT-PCR of embryonic and adult human tissues, hNAT3 was detected to be predominantly expressed in the liver and to a much lesser extent in the muscle, kidney, and pancreas. The data obtained in this study are likely to offer critical clues for identification of amino acid transporter-associated diseases. PMID- 11414755 TI - Genetic analysis of a new mouse model for non-insulin-dependent diabetes. AB - The TallyHo (TH) mouse strain is a newly established model for non-insulin dependent diabetes mellitus (NIDDM). TH mice show obesity, hyperinsulinemia, hyperlipidemia, and male-limited hyperglycemia. A genetic dissection of the diabetes syndrome has been carried out using male backcross 1 progeny obtained from crosses between (C57BL/6J x TH)F1 and TH mice or (CAST/Ei x TH)F1 and TH mice. A genome-wide scan reveals three quantitative trait loci (QTLs), Tanidd1-3 (TH-associated NIDDM) linked to hyperglycemia. The major QTL (common in both crosses), Tanidd1, maps to chromosome (Chr) 19. Additionally, gene-gene interactions contributing to hyperglycemia have been observed between Tanidd1 and a locus on Chr 18 as well as between Tanidd2 and a locus on Chr 16. The overt hyperglycemia in TH mice is, therefore, likely due to a mutation in a major diabetes susceptibility locus on Chr 19, which interacts with additional genes to lead to an observable phenotype. PMID- 11414756 TI - Rat-mouse and rat-human comparative maps based on gene homology and high resolution zoo-FISH. AB - The laboratory rat, Rattus norvegicus, and the laboratory mouse, Mus musculus, are key animal models in biomedical research. A deeper understanding of the genetic interrelationsships between Homo sapiens and these two rodent species is desirable for extending the usefulness of the animal models. We present comprehensive rat-human and rat-mouse comparative maps, based on 1090 gene homology assignments available for rat genes. Radiation hybrid, FISH, and zoo FISH mapping data have been integrated to produce comparative maps that are estimated to comprise 83-100% of the conserved regions between rat and mouse and 66-82% of the conserved regions between rat and human. The rat-mouse zoo-FISH analysis, supported by data for individual genes, revealed nine previously undetected conserved regions compared to earlier reports. Since there is almost complete genome coverage in the rat-mouse comparative map, we conclude that it is feasible to make accurate predictions of gene positions in the rat based on gene locations in the mouse. PMID- 11414757 TI - Molecular basis of mucopolysaccharidosis type IIIB in emu (Dromaius novaehollandiae): an avian model of Sanfilippo syndrome type B. AB - Sanfilippo syndrome type B, or mucopolysaccharidosis (MPS) IIIB, is an autosomal recessive disease caused by a deficiency of lysosomal alpha-N acetylglucosaminidase (NAGLU). In Dromaius novaehollandiae (emu), a progressive neurologic disease was recently discovered, which was characterized by NAGLU deficiency and heparan sulfate accumulation. To define the molecular basis, the sequences of the normal emu NAGLU cDNA and gene were determined by PCR-based approaches using primers for highly conserved regions of evolutionarily distant NAGLU homologues. It was observed that the emu NAGLU gene is structurally similar to that of human and mouse, but the introns are considerably shorter. The cDNA had an open reading frame (ORF) of 2259 bp. The deduced amino acid sequence is estimated to share 64% identity with human, 63% with mouse, 41% with Drosophila, 39% with tobacco, and 35% with the Caenorhabditis elegans enzyme. Three normal and two affected emus were studied for nucleotide sequence covering the entire coding region and exon-intron boundaries. Unlike the human gene, emu NAGLU appeared to be highly polymorphic: 19 variations were found in the coding region alone. The two affected emus were found to be homozygous for a 2-bp deletion, 1098-1099delGG, in exon 6. The resulting frameshift predicts a longer ORF of 2370 bp encoding a polypeptide with 37 additional amino acids and 387 altered amino acids. The availability of mutation screening in emus now permits early detection of MPS IIIB in breeding stocks and is an important step in characterizing this unique, naturally occurring avian model for the development of gene transfer studies. PMID- 11414758 TI - Electroconvulsive thresholds of inbred mouse strains. AB - The electroconvulsive threshold (ECT) test is used commonly in the screening of anti-epileptic drugs in rodent models, but little is known about its genetic or mechanistic basis. Thresholds for minimal clonic, maximal tonic, or psychomotor (partial) seizures were determined in 16 different inbred mouse strains in two different laboratories. A wide range of thresholds was observed, suggesting that a variety of neuroexcitability alleles exist in inbred strains. Although there was generally good cross-strain correlation between the three seizure types, several outlier strains were detected, showing that genetically encoded differences can affect the ability of a particular seizure type to spread through the brain. Furthermore, the relative seizure susceptibility of a strain was comparable between the two laboratories, suggesting that despite different test sites, instrumentation, and personnel, the ECT assay is portable and that common inbred strains can often be relied upon as calibration standards. Last, the ECT paradigm was also sensitive enough to detect single locus differences, laying the groundwork for mutation screens for new neuroexcitability models. PMID- 11414759 TI - A 76-bp deletion in the Mip gene causes autosomal dominant cataract in Hfi mice. AB - Hfi is a dominant cataract mutation where heterozygotes show hydropic lens fibers and homozygotes show total lens opacity. The Hfi locus was mapped to the distal part of mouse chromosome 10 close to the major intrinsic protein (Mip), which is expressed only in cell membranes of lens fibers. Molecular analysis of Mip revealed a 76-bp deletion that resulted in exon 2 skipping in Mip mRNA. In Hfi/Hfi this deletion resulted in a complete absence of the wildtype Mip. In contrast, Hfi/+ animals had the same amount of wildtype Mip as +/+. Results from pulse-chase expression studies excluded hetero-oligomerization of wildtype and mutant Mip as a possible mechanism for cataract formation in the Hfi/+. We propose that the cataract phenotype in the Hfi heterozygote mutant is due to a detrimental gain of function by the mutant Mip resulting in either cytotoxicity or disruption in processing of other proteins important for the lens. Cataract formation in the Hfi/Hfi mouse is probably a combined result of both the complete loss of wildtype Mip and a gain of function of the mutant Mip. PMID- 11414760 TI - Isolation and characterization of a novel gene from the DiGeorge chromosomal region that encodes for a mediator subunit. AB - Hemizygous deletions on chromosome 22q11.2 result in developmental disorders referred to as DiGeorge syndrome (DGS)/velocardiofacial syndrome (VCFS). We report the isolation of a novel gene, PCQAP (PC2 glutamine/Q-rich-associated protein), that maps to the DiGeorge typically deleted region and encodes a protein identified as a subunit of the large multiprotein complex PC2. PC2 belongs to the family of the human Mediator complexes, which exhibit coactivator function in RNA polymerase II transcription. Furthermore, we cloned the homologous mouse Pcqap cDNA. There is 83% amino acid identity between the human and the mouse predicted protein sequences, with 96% similarity at the amino- and carboxy-terminal ends. To assess the potential involvement of PCQAP in DGS/VCFS, its developmental expression pattern was analyzed. In situ hybridization of mouse embryos at different developmental stages revealed that Pcqap is ubiquitously expressed. However, higher expression was detected in the frontonasal region, pharyngeal arches, and limb buds. Moreover, analysis of subjects carrying a typical 22q11 deletion revealed that the human PCQAP gene was deleted in all patients. Many of the structures affected in DGS/VCFS evolve from Pcqap expressing cells. Together with the observed haploinsufficiency of PCQAP in DGS/VCFS patients, this finding is consistent with a possible role for this novel Mediator subunit in the development of some of the structures affected in DGS/VCFS. PMID- 11414761 TI - Homologues to the first gene for autosomal dominant polycystic kidney disease are pseudogenes. AB - PKD1 is the first gene identified to be causative for the condition of autosomal dominant polycystic kidney disease. There are several genes homologous to PKD1 that are located proximal to the master gene on the same chromosome. Two of these genes have been recently covered in a large sequencing work on chromosome 16, and their structure has been broadly analyzed. However, the major question whether homologous genes (HG) code for functionally active polypeptides has not been resolved so far. The current study identifies and partially characterizes four more homologues of PKD1, different from the previously published sequence, two of which were found by screening of a BAC library and the other two contained in available databases. Analysis of HG transcripts shows that they are not translated in the model cell line T98G. Taken together, these findings suggest that homologues to PKD1 form a family of pseudogenes. PMID- 11414762 TI - Mepe, the gene encoding a tumor-secreted protein in oncogenic hypophosphatemic osteomalacia, is expressed in bone. AB - The MEPE (matrix extracellular phosphoglycoprotein) gene is a strong candidate for the tumor-derived phosphaturic factor in oncogenic hypophosphatemic osteomalacia (OHO). X-linked hypophosphatemia (XLH) is phenotypically similar to OHO and results from mutations in PHEX, a putative metallopeptidase believed to process a factor(s) regulating bone mineralization and renal phosphate reabsorption. Here we report the isolation of the murine homologue of MEPE, from a bone cDNA library, that encodes a protein of 433 amino acids, 92 amino acids shorter than human MEPE. Mepe, like Phex, is expressed by fully differentiated osteoblasts and down-regulated by 1,25-(OH)2D3. In contrast to Phex, Mepe expression is markedly increased during osteoblast-mediated matrix mineralization. Greater than normal Mepe mRNA levels were observed in bone and osteoblasts derived from Hyp mice, the murine homologue of human XLH. Our data provide the first evidence that MEPE/Mepe is expressed by osteoblasts in association with mineralization. PMID- 11414763 TI - Mutation analyses of 268 candidate genes in human tumor cell lines. AB - We have performed a homozygous deletion screen on 268 candidate genes in 90 human tumor cell lines derived from multiple types of cancers. Most of the candidate genes investigated have been proposed to be involved in cellular processes that are germane to cancer progression, such as cell cycle control, genome maintenance, chromatin remodeling, cell adhesion, and apoptosis. We have detected novel homozygous deletions affecting four independent loci: Brahma-related gene (SMARCA4) on chromosome 19p in the TSU-Pr1 prostate and A427 lung carcinoma lines, Map Kinase Kinase 3 (MAP2K3) on 17q in the NCI-H774 lung tumor cell line, TMPRSS2 on 21q in the Bx PC-3 pancreatic carcinoma line, and Cadherin 6 (CDH6) on 5p in the SK-LU-1 lung carcinoma line. Subsequent analyses of the coding sequences of these four genes using cDNAs from a panel of tumor cell lines revealed multiple sequence variants. The results of this mutation study serve to demonstrate the feasibility of performing high-throughput screens of candidate genes in tumor cell lines to identify genes that may be targeted for mutation during the development of cancer. PMID- 11414764 TI - Quantitative trait locus analysis for chronic pancreatitis and diabetes mellitus in the WBN/Kob rat. AB - A causative gene mutation is still undefined in approximately half of patients with hereditary pancreatitis, and no genetic factor has been identified in most patients with sporadic chronic pancreatitis. To identify a pancreatitis associated gene, we performed a quantitative trait locus (QTL) analysis for the traits of chronic pancreatitis and diabetes mellitus in WBN/Kob rats. We identified two highly significant QTLs for chronic pancreatitis and/or hyperinsulinemia on chromosomes 7 and X. These QTLs were located on completely different chromosomal regions from those of causative genes that have been reported for human chronic pancreatitis: PRSS1, CFTR, and SPINK1. For these QTLs, prevalences of the WBN/Kob allele significantly increased in the rats with chronic pancreatitis. These findings indicate that chronic pancreatitis in WBN/Kob rats is controlled by multiple genes, and a genetic analysis in WBN/Kob rats might be useful for gene targeting for human chronic pancreatitis. PMID- 11414765 TI - Imprinting status of 11p15 genes in Beckwith-Wiedemann syndrome patients with CDKN1C mutations. AB - Beckwith-Wiedemann syndrome (BWS) is an imprinting disorder characterized by somatic overgrowth, congenital malformations, and predisposition to childhood tumors. Aberrant expression of multiple imprinted genes, including H19, IGF2, KCNQ1OT1, and CDKN1C, has been observed in BWS patients. It has been estimated that mutations in CDKN1C occur in 12-17% of BWS patients. We have screened 10 autosomal dominant pedigrees and 65 sporadic BWS cases by PCR/heteroduplex analysis and DNA sequencing and have identified four mutations, two of which were associated with biallelic IGF2 expression and normal H19 and KCNQ1OT1 imprinting. One patient demonstrated phenotypic expression of paternally transmitted mutation in this maternally expressed gene, a second proband is the child of one of a pair of monozygotic twin females who carry the mutation de novo, and a third patient exhibited unusual skeletal changes more commonly found in other overgrowth syndromes. When considered with other studies published to date, this work reveals the frequency of CDKN1C mutations in BWS to be only 4.9%. This is the first report of an analysis of the imprinting status of genes in the 11p15 region where CDKN1C mutations were associated with loss of IGF2 imprinting and maintenance of H19 and KCNQ1OT1 imprinting. PMID- 11414766 TI - Cloning and characterization of the human retina-specific gene MPP4, a novel member of the p55 subfamily of MAGUK proteins. AB - To identify novel retina-specific genes systematically, we are performing expression profiling of retina ESTs that have been assembled in the human UniGene clusters. In this study, we report the 2619-bp full-length cDNA cloning and genomic organization of a gene corresponding to an EST cluster that was demonstrated to be exclusively present in retinal tissue. Alignment of the deduced amino acid sequence to sequence from protein databases revealed this gene, termed MPP4, to be a member of the membrane-associated guanylate kinase (MAGUK) protein family. It consists of 637 amino acids and contains the characteristic MAGUK motifs: an N-terminal PDZ domain, a central src homology 3 region (SH3), and a C-terminal guanylate kinase-like (GUK) domain. Due to the presence of only one PDZ motif, MPP4 is part of the p55 subfamily, named after the major palmitoylated erythrocyte membrane protein p55/MPP1. MAGUK proteins serve as molecular scaffolds to coordinate the membrane-associated cytoskeleton, ion channel and receptor clustering, signaling pathways, and the formation of cellular junctions. The abundant expression of MPP4 in the human retina suggests an important but so far unknown function in this tissue. Colocalization of MPP4 and autosomal recessive retinitis pigmentosa 26 (RP26) on chromosome 2q31-q33 makes this transcript an attractive candidate for the disease gene. PMID- 11414767 TI - Molecular cloning of a novel human acid phosphatase gene (ACPT) that is highly expressed in the testis. AB - Acid phosphatases are enzymes capable of hydrolyzing orthophosphoric acid esters in an acid medium. Prostatic acid phosphatase has served as a tumor marker for metastatic prostate cancer for many years. We have cloned a new human acid phosphatase gene (named testicular acid phosphatase, ACPT), which is expressed mainly in testis and to a lower extent in the prostate, trachea, and other tissues. This gene maps to chromosome 19q13.4, in an area that harbors many cancer-related genes. The testicular acid phosphatase gene is composed of 11 exons, and the protein is predicted to have a luminal domain, a transmembrane domain, and a cytoplasmic domain. The N-terminal end of the protein encodes a signal peptide. The protein has approximately 50% homology with both the prostatic and the lysosomal acid phosphatases, and the position of the cysteine residues, the N-glycosylation sites, and the histidine catalytic site are conserved among the three proteins. The testicular acid phosphatase gene is up regulated by androgens and is down-regulated by estrogens in the prostate cancer cell line LNCaP. Our preliminary results indicate that this gene exhibits a lower level of expression in testicular cancer tissues than in their normal counterparts. PMID- 11414769 TI - Simultaneous identification of mutations by dual-parameter multiplex hybridization in peptide nucleic acid-containing virtual arrays. AB - The physical entrapment of peptide nucleic acids (PNA) in electrophoresis media provides a system for performing real-time hybridization. DNA strands fully complementary to the target PNA are retarded compared to single-nucleotide mismatched strands. A second parameter, that of amplicon length, has been introduced to perform multiplex analyses on several mutations simultaneously. Size fractionation creates a virtual array of PCR products that can hybridize to one of a set of mutation-specific PNAs present within the matrix. Each targeted mutation can be identified by the size of its corresponding amplicon. Its genotype is characterized by its interaction with a specific PNA that gives a visually resolved distinction between wildtype and mutant allele. In contrast to conventional hybridization, heterozygotes are readily distinguished from homozygotes. Using a capillary electrophoresis-based DNA sequencer, this approach has been used to automate the identification of the H63D, S65C, and C282Y mutations in the hereditary hemochromatosis gene. PMID- 11414768 TI - Genomic organization and isoform-specific tissue expression of human NAPOR (CUGBP2) as a candidate gene for familial arrhythmogenic right ventricular dysplasia. AB - Neuroblastoma apoptosis-related RNA-binding protein (NAPOR; HGMW-approved symbol CUGBP2) is a newly discovered gene prominently induced during apoptosis, suggesting that it plays a role during apoptosis. We have found that it is encoded by a gene located on chromosome 10p13-p14 between Genethon markers D10S547 and D10S223, a region to which we have recently localized a gene responsible for arrhythmogenic right ventricular dysplasia (ARVD). To examine its possible role in the pathogenesis of ARVD, we determined the genomic organization of the human NAPOR gene including its exon-intron boundaries and the putative promoter sequence, which provide a plausible mechanism for its alternative mRNA splicing. We also demonstrated that three isoforms of the NAPOR transcript were differently expressed, with NAPOR-3 being nearly neuron specific while the other two forms were ubiquitously expressed. The expression of NAPOR is differentially regulated during development. Finally, we screened the members of the ARVD family for mutations and identified two DNA sequence variants in the protein-coding exons of NAPOR, neither of which was responsible for ARVD. While the function of NAPOR remains to be elucidated, our current characterization of the NAPOR gene will be valuable for further clinical and functional study. PMID- 11414770 TI - Cloning, expression patterns, and chromosome localization of three human and two mouse homologues of GABA(A) receptor-associated protein. AB - Type A receptors of gamma-aminobutyric acid (GABA), an inhibitory neurotransmitter, contain alpha, beta, delta, gamma, and rho subunits. The gamma subunit has four subtypes: gamma1, gamma2, gamma3, andgamma4. GABA(A) receptor associated protein (GABARAP) was previously demonstrated to act as a linker protein between microtubules and the gamma2 subunit of GABA(A) receptors. However, no other linker proteins have been identified as mediating the linkage of microtubules and the remaining subunits of GABA(A) receptors. In this study we identified three human paralogues (GABARAPL1, GABARAPL2, and GABARAPL3) and two mouse orthologues (Gabarapl1 and Gabarapl2) of human GABARAP, all of which encoded 117 amino acids, as does Gabarapl. The expression patterns of GABARAPL1, GABARAPL2, and GABARAP in 16 adult tissues showed that they were expressed ubiquitously. The expression levels of GABARAPL1 as a 2.3-kb transcript were very high in brain, heart, peripheral blood leukocytes, liver, kidney, placenta, and skeletal muscle, very low in thymus and small intestine, and moderate in other tissues tested. The unique 1.35-kb transcript of GABARAPL2 was expressed at high levels in heart, brain, testis, prostate, ovary, spleen, and skeletal muscle, at very low levels in lung, thymus, and small intestine, and moderately in other tissues tested. For GABARAP, a 1.3-kb transcript was abundantly expressed in all tested tissues with small variation. The expression patterns of Gabarapl1 and Gabarapl2 were similar to those of their counterparts in human. In addition, GABARAPL1 was localized to human chromosome 12p12.3 and GABARAPL2 to 16q22.3 q24.1 by RH mapping, while GABARAP and GABARAPL3 were found to be localized at chromosomes 17p13.2 and 15q25.1, respectively, by searching the related databases. Sequence comparison of the cDNAs and their corresponding genomic sequences shows that GABARAP, GABARAPL1, and GABARAPL2 are composed of four exons each, while GABARAPL3 is distributed only at one exon. PMID- 11414771 TI - Comparative context of Plio-Pleistocene hominin brain evolution. AB - One of the distinguishing features of Homo sapiens is its absolutely and relatively large brain. This feature is also seen in less extreme form in some fossil Homo species. However, are increases in brain size during the Plio Pleistocene only seen in Homo, and is brain enlargement among Plio-Pleistocene primates confined to hominins? This study examines evidence for changes in brain size for species and lineage samples of three synchronic East African fossil primate groups, the two hominin genera Homo and Paranthropus, and the cercopithecoid genus Theropithecus. Hominin endocranial capacity data were taken from the literature, but it was necessary to develop an indirect method for estimating the endocranial volume of Theropithecus. Bivariate and multivariate regression equations relating measured endocranial volume to three external cranial dimensions were developed from a large (ca. 340) sample of modern African cercopithecoids. These equations were used to estimate the endocranial volumes of 20 Theropithecus specimens from the African Plio-Pleistocene. Spearman's rho and the Hubert nonparametric test were used to search for evidence of temporal trends in both the hominin and Theropithecus data. Endocranial volume apparently increased over time in both Homo and Paranthropus boisei, but there was no evidence for temporal trends in the endocranial volume of Theropithecus. Thus, hypotheses which suggest a mix of environmental, social, dietary, or other factors as catalysts for increasing brain in Plio-Pleistocene primates must accommodate evidence of brain enlargement in both Homo and Paranthropus, and explain why this phenomenon appears to be restricted to hominins. PMID- 11414772 TI - New hominids from the Lake Turkana Basin, Kenya. AB - New hominid fossils from the Lake Turkana Basin range in age from ca. 3.35 to ca. 1.0 Ma. Those recovered from sediments stratigraphically just above the Tulu Bor Tuff in the Lomekwi Member of the Nachukui Formation are best attributed to Australopithecus afarensis. This species is rare in Kenya, probably because of the scarcity of sediments deposited during its time span. Younger specimens are referable either to the megadont A. boisei or early Homo. Collectively the new fossils promote further understanding of morphological variation in East African Plio-Pleistocene hominids. PMID- 11414773 TI - Phylogenetic relationships among the macaques: evidence from the nuclear locus NRAMP1. AB - The macaques, genus Macaca, represent one of the most successful radiations within the Order Primates, with a geographical distribution that ranks second in size only to that of humans among extant primates. Although the number of macaque species recognized depends on the classification scheme used, most authors currently follow the classifications of either Fooden or Delson, both of whom recognize 19 extant macaque species. These two classifications differ in their placement of macaque species into more inclusive taxa (i.e., species groups). While researchers have attempted to use mitochondrial DNA (mtDNA) to resolve these phylogenetic relationships, different studies have generated conflicting conclusions. Consequently, we screened nuclear DNA sequences of a large number of macaques to determine if such data provide greater insight into the phylogenetic relationships among macaques. The data generated from the comparison of two (noncoding) introns within the natural resistance-associated macrophage protein 1 (NRAMP1) gene generally agree with the classification scheme of Delson. However, the data also support several novel observations. Specifically, the NRAMP1 data demonstrate that M. silenus and M. nemestrina lack nuclear genetic variation, while M. assamensis and M. radiata exhibit the greatest amount of genetic variation. In addition, these data suggest that M. fascicularis may not be as "primitive" (with respect to other members of the fascicularis group) as the mtDNA based data suggest. PMID- 11414774 TI - Delivery of musculoskeletal therapy on line? PMID- 11414775 TI - Chiropractic manipulation for the foot: Diversified chiropractic techniques. AB - There has been increasing acceptance and development of manual methods in providing for the needs of patients with musculoskeletal dysfunction. Several professions have helped fuel this growth, including the chiropractic profession. To date, there has been only a small amount of collaboration between chiropractors and physical therapists. This paper provides a base foundation for one small part of general chiropractic practice, i.e. procedures used for manipulating the foot. Information is provided about the specific diagnostic procedures used by the chiropractic profession in assessing the joints and soft tissues of the foot, followed by descriptions of a number of chiropractic manipulative techniques drawn from the form of chiropractic in widest usage, Diversified technique. For each technique, information is provided on indications for use, patient position, therapist position, hand placements and procedure. In addition, a short discussion on the genesis of Diversified technique is provided. PMID- 11414776 TI - Cervical mobilisation: concurrent effects on pain, sympathetic nervous system activity and motor activity. AB - Recent findings that spinal manual therapy (SMT) produces concurrent hypoalgesic and sympathoexcitatory effects have led to the proposal that SMT may exert its initial effects by activating descending inhibitory pathways from the dorsal periaqueductal gray area of the midbrain (dPAG). In addition to hypoalgesic and sympathoexcitatory effects, stimulation of the dPAG in animals has been shown to have a facilitatory effect on motor activity. This study sought to further investigate the proposal regarding SMT and the PAG by including a test of motor function in addition to the variables previously investigated. Using a condition randomised, placebo-controlled, double blind, repeated measures design, 30 subjects with mid to lower cervical spine pain of insidious onset participated in the study. The results indicated that the cervical mobilisation technique produced a hypoalgesic effect as revealed by increased pressure pain thresholds on the side of treatment (P=0.0001) and decreased resting visual analogue scale scores (P=0.049). The treatment technique also produced a sympathoexcitatory effect with an increase in skin conductance (P<0.002) and a decrease in skin temperature (P=<0.02). There was a decrease in superficial neck flexor muscle activity (P<0.0002) at the lower levels of a staged cranio-cervical flexion test. This could imply facilitation of the deep neck flexor muscles with a decreased need for co-activation of the superficial neck flexors. The combination of all findings would support the proposal that SMT may, at least initially, exert part of its influence via activation of the PAG. PMID- 11414777 TI - Abdominal muscle activity during the abdominal hollowing manoeuvre in the four point kneeling and prone positions. AB - This study investigated the pattern of recruitment of the abdominal muscles during the abdominal hollowing manoeuvre (AHM), in four point kneeling and prone. Muscle activity was detected and recorded by surface electromyography (EMG), in 20 healthy subjects, 8 male and 12 female, aged between 19 and 30 during both maximal activity, and the AHM. The degree of activation during hollowing was expressed as a percentage of maximal. There was activity in the internal abdominal oblique (IO) during all hollowing manoeuvres, in both starting positions, whilst only two subjects showed any activity in rectus abdominis (RA). Simultaneous activity in IO and EO occurred both in four point kneeling (45% of the subjects) and especially in prone (75% of the subjects). Across both positions IO activity was significantly greater than EO. There was no significant difference in EO or IO activity comparing across positions. These results suggest that the AHM when performed in four point kneeling and prone always activates IO and rarely activates RA, whilst elimination of activity in EO is more difficult to achieve, particularly in prone. Isolated activity in the deep abdominal muscles may not always be achievable in these starting positions. PMID- 11414778 TI - Shoulder girdle elevation during neurodynamic testing: an assessable sign? AB - One of the signs advocated for monitoring during neurodynamic testing in the assessment of patients with upper quadrant disorders, is the response of the shoulder girdle. It is stated that a protective rising of the shoulder girdle is present when patients with neurogenic disorders are assessed and that the elevation is absent in asymptomatic subjects. As sensory responses are elicited in the majority of asymptomatic subjects and as the range of motion (ROM) is often limited during neurodynamic testing, it is questionable whether the elevation of the shoulder girdle would be absent in asymptomatic subjects. The aim of this study was to measure the shoulder girdle elevation force during five variants of the neural tissue provocation test for the median nerve. Thirty-five asymptomatic male subjects were assessed. A load cell was used to measure the amount of shoulder girdle elevation force and two electrogoniometers were used to measure the ROM at the elbow and wrist. When the ROM at the end of the test was restricted, a gradual increase in shoulder girdle elevation force could be observed throughout the test. Compared to the initial force at the start of the test, all variants resulted in a significant increase in force. It is concluded that a gradual increase in shoulder girdle elevation force should not be regarded as an abnormal sign in the interpretation of neurodynamic tests. PMID- 11414779 TI - Intermittent isometric fatigue study of the lumbar multifidus muscle in four point kneeling: an intra-rater reliability investigation. AB - Back muscle endurance is considered important in low back pain (LBP) rehabilitation. Specific training of multifidus may also be necessary to restore normal low back function. The reliability of surface electromyogram (EMG) to assess endurance of the multifidus muscle during intermittent isometric exercise was evaluated. Multifidus endurance was monitored in the four-point kneeling exercise position using the power spectral analysis method. Twenty healthy volunteers were tested on three separate occasions. Subjects performed repeat 10 s high-intensity voluntary contractions of multifidus for 3 min. The median frequency (MF) and the integrated-rectified (I-R) EMG signal displayed the fatigue pattern of multifidus. Intraclass correlation coefficients indicated fair good reproducibility for MF (0.48-0.67) but poor reliability for IR-EMG. In conclusion, problems concerning functional testing protocols for the back muscles remain and careful development is necessary for more realistic rehabilitation monitoring. PMID- 11414780 TI - Acute low cervical nerve root conditions: symptom presentations and pathobiological reasoning. AB - Acute low cervical nerve root conditions may be easily misdiagnosed. The perspective presented is that their symptom presentation is not as straightforward as the classic descriptions of brachialgia would have us believe. This clinical commentary presents a series of observations and reasoning models that are relevant to patient symptom presentations believed to be of cervical nerve root origin. Clinicians are urged to consider low cervical nerve root assessment in the light of our current understanding of neural sensitivity, pain science, nerve root biomechanics and the presence and effect of degenerative changes. This particularly relates to thoughts about cervical movements and postures being able to bring forces to bear on nerve roots via compressive as well as elongation forces. PMID- 11414781 TI - The locked lumbar facet joint: intervention using mobilizations with movement. PMID- 11414782 TI - Bibliography. PMID- 11414784 TI - Chick ciliary neurotrophic factor is secreted via a nonclassical pathway. AB - In contrast to mammalian ciliary neurotrophic factors (CNTFs), chick CNTF is secreted, although it lacks an N-terminal signal. We determined that a 52 aa region of chick CNTF containing an internal hydrophobic domain could direct secretion of rat CNTF. Using a stable cell line that overexpressed chick CNTF, we found that chick CNTF immunoreactivity was punctate throughout the cytosol. Cellular fractionation confirmed chick CNTF to be protected by vesicles. Chick CNTF did not colocalize with fibronectin, calreticulin, wheat germ agglutinin binding sites, or with transferrin receptor. The distribution of chick CNTF was altered neither by brefeldin A nor by chloroquine treatment. Although the punctate pattern of chick CNTF immunoreactivity was not due to reuptake, chick CNTF could be found in a cellular compartment labeled after a brief incubation with dextran microbeads. When synthesized in vitro, chick CNTF did not translocate into microsomes. We conclude that chick CNTF is secreted via a nonclassical pathway. PMID- 11414785 TI - MMP-related gelatinase activity is strongly induced in scar tissue of injured adult spinal cord and forms pathways for ingrowing neurites. AB - Scar formation following adult spinal cord (SC) hemisection is accompanied by important remodeling of the surrounding extracellular matrix (ECM). Since ECM molecules provide the substrate for axon growth, these changes in ECM composition are likely to influence the process of axonal regeneration. Here we investigated whether scar formation could be associated with the activation of matrix metalloproteinases (MMPs), a class of proteins implicated in ECM remodeling thought to favor axonal regeneration in the peripheral nervous system. Two members of the MMP family, MMP-2 and MMP-9, were found to be transiently upregulated in the SC wound. In situ fluorescent zymography revealed a MMP related gelatinase activity (GA) in the wound, which was spatially and temporally correlated with scar formation. The GA formed a striking pattern of interwoven pathways along which neurites were seen to grow. These pathways corresponded to the distribution of other ECM molecules, which are known to have antagonistic effects on axonal regrowth. Our results suggest that neurite ingrowth into the wound may transiently benefit from this ECM remodeling and, in particular, from the upregulation of MMPs. PMID- 11414786 TI - Bidirectional regulation of neurite elaboration by alternatively spliced metabotropic glutamate receptor 5 (mGluR5) isoforms. AB - Alternative splicing in the mGluR5 gene generates two different receptor isoforms, of which expression is developmentally regulated. However, little is known about the functional significance of mGluR5 splice variants. We have examined the functional coupling, subcellular targeting, and effect on neuronal differentiation of epitope-tagged mGluR5 isoforms by expression in neuroblastoma NG108-15 cells. We found that both mGluR5 splice variants give rise to comparable [Ca2+]i transients and have similar pharmacological profile. Tagged receptors were shown by immunofluorescence to be inserted in the plasma membrane. In undifferentiated cells the subcellular localization of the two mGluR5 isoforms was partially segregated, whereas in differentiated cells the labeling largely redistributed to the newly formed neurites. Interestingly, we demonstrate that mGluR5 splice variants dramatically influence the formation and maturation of neurites; mGluR5a hinders the acquisition of mature neuronal traits and mGluR5b fosters the elaboration and extension of neurites. These effects are partly inhibited by MPEP. PMID- 11414787 TI - Gephyrin-independent clustering of postsynaptic GABA(A) receptor subtypes. AB - Gephyrin has been shown to be essential for the synaptic localization of the inhibitory glycine receptor and major GABA(A) receptor (GABA(A)R) subtypes. However, in retina certain GABA(A)R subunits are found at synaptic sites in the absence of gephyrin. Here, we quantitatively analyzed GABA(A)R alpha1, alpha2, alpha3, alpha5, beta2/3, and gamma2 subunit immunoreactivities in spinal cord sections derived from wild-type and gephyrin-deficient (geph -/-) mice. The punctate staining of GABA(A)R alpha1 and alpha5 subunits was unaltered in geph -/ mice, whereas the numbers of alpha2-, alpha3-, beta2/3-, and gamma2-subunit immunoreactive synaptic sites were significantly or even strikingly reduced in the mutant animals. Immunostaining with an antibody specific for the vesicular inhibitory amino acid transporter revealed that the number of inhibitory presynaptic terminals is unaltered upon gephyrin deficiency. These data show that in addition to gephyrin other clustering proteins must exist that mediate the synaptic localization of selected GABA(A)R subtypes. PMID- 11414788 TI - Migration and multipotentiality of PSA-NCAM+ neural precursors transplanted in the developing brain. AB - By optimizing the previously described strategy for obtention of spheres enriched in PSA-NCAM+ precursors, we prepared PSA-NCAM-immunoselected cell populations from cerebral hemispheres of neonatal MBP-LacZ transgenic mice. These cells expressed Nestin, exhibited clonal expansion potential and formed spheres, which were initially enriched in PSA-NCAM+ cells but became enriched in GD3+ oligodendrocyte progenitors after 1 week in B104 contionned medium. One month after their periventricular transplantation into the brain of wild-type and/or shiverer newborn mice, cells from PSA-NCAM+ spheres exhibited a higher rostral migration potential than cells from GD3+ spheres, and clearly contributed to myelination in the olfactory bulb. In shiverer hosts, both sphere populations generated oligodendrocytes with similar myelination potential. In addition PSA NCAM+ sphere cells generated GFAP+ astrocytes and NeuN+ neurons, depending on their site of insertion. These results evidence the high plasticity of newborn PSA-NCAM+ neural precursors and suggest that they are promising tools for cell therapy of CNS diseases, including myelin disorders. PMID- 11414789 TI - Platelet-derived growth factor receptor-alpha in ventricular zone cells and in developing neurons. AB - Cells in the early neuroepithelium differentiate and give rise to all cells in the central nervous system (CNS). The ways from a multipotent CNS stem cell to specialized neurons and glia are not fully understood. Using immunohistochemistry we found that neuroepithelial cells express the platelet-derived growth factor receptor-alpha (PDGFR-alpha) in the neural plate at embryonic day 8.5 and onwards in the neural tube. The protein was polarized to ventricular endfeet. Furthermore, PDGFR-alpha expression was localized to cells undergoing early neuronal development. We also found PDGFR-alpha expression in developing granule cells in the postnatal cerebellum, in Purkinje cells in the adult cerebellum and on processes of developing dorsal root ganglion cells. Previous reports mainly describe PDGFR-alpha expression in oligodendrocyte precursors and glial cells. We believe, in line with a few previous reports, that the PDGFR-alpha in addition marks a pool of undifferentiated cells, which are able to differentiate into neurons. PMID- 11414790 TI - Enhanced hippocampal long-term potentiation in mice lacking heparin-binding growth-associated molecule. AB - Heparin-binding growth-associated molecule (HB-GAM) (pleiotrophin) is a highly conserved extracellular matrix-associated protein implicated in a diverse range of developmental processes, including the formation and plasticity of neuronal connections. Using gene targeting, we have in the present study created HB-GAM deficient mice that are viable and fertile and show no gross anatomical abnormalities. The hippocampal structure as well as basal excitatory synaptic transmission in the area CA1 appear normal in the mice lacking HB-GAM. However, hippocampal slices from HB-GAM-deficient mice display a lowered threshold for induction of long-term potentiation (LTP), which reverts back to the wild-type level by application of HB-GAM. HB-GAM expression in hippocampus is activity dependent and upregulated in several neuropathological conditions. Thus, we suggest that HB-GAM acts as an inducible signal to inhibit LTP in hippocampus. PMID- 11414791 TI - High abundance of GluR1 mRNA and reduced Q/R editing of GluR2 mRNA in individual NADPH-diaphorase neurons. AB - Striatal and cortical neurons containing NADPH-diaphorase [NADPH-d(+)] are highly vulnerable to excitotoxicity that is induced by activation of alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)- or kainate-sensitive glutamate receptors. This has been attributed to Ca2+ entry through AMPA/kainate receptors in NADPH-d(+) neurons. In this study, we applied single cell RT-PCR technique to test the hypothesis that differences in levels and processing of the GluR2 subunit would contribute to the selective vulnerability of NADPH-d(+) neurons to AMPA. The nested PCR specific for GluR1-GluR4 showed that rat striatal NADPH-d(+) neurons expressed twice as much GluR1 mRNA as NADPH-d(-) neurons did. The percentage of RNA editing at the Q/R site of GluR2 was 46% in NADPH-d(+) neurons and 92% in NADPH-d(-) neurons. These results suggest that the unedited expression of GluR2 and the reduced ratio of GluR2/GluR1 render NADPH-d(+) neurons highly sensitive to Ca2+-mediated AMPA neurotoxicity. In support of this, most NADPH d(+) neurons exposed to 100 microM AMPA showed Co2+ uptake and survived AMPA challenge only in the absence of extracellular Ca2+. PMID- 11414792 TI - Expression of Eph receptors in skeletal muscle and their localization at the neuromuscular junction. AB - The participation of ephrins and Eph receptors in guiding motor axons during muscle innervation has been well documented, but little is known about their expression and functional significance in muscle at later developmental stages. Our present study investigates the expression and localization of Eph receptors and ephrins in skeletal muscle. Prominent expression of EphA4, EphA7, and ephrin A ligands was detected in muscle during embryonic development. More importantly, both EphA4 and EphA7, as well as ephrin-A2, were localized at the neuromuscular junction (NMJ) of adult muscle. Despite their relative abundance, they were not localized at the synapses during embryonic stages. The concentration of EphA4, EphA7, and ephrin-A2 at the NMJ was observed at postnatal stages and the synaptic localization became prominent at later developmental stages. In addition, expression of Eph receptors was increased by neuregulin and after nerve injury. Furthermore, we demonstrated that overexpression of EphA4 led to tyrosine phosphorylation of the actin-binding protein cortactin and that EphA4 was coimmunoprecipitated with cortactin in muscle. Taken together, our findings indicate that EphA4 is associated with the actin cytoskeleton. Since actin cytoskeleton is critical to the formation and stability of NMJ, the present findings raise the intriguing possibility that Eph receptors may have a novel role in NMJ formation and/or maintenance. PMID- 11414793 TI - Sensory axon response to substrate-bound Slit2 is modulated by laminin and cyclic GMP. AB - In vertebrates, Slit2 is a chemorepellent for some developing axons but stimulates axonal elongation and branching of sensory axons. In vivo, Slit2 is cleaved into 140-kDa N-terminal (Slit2-N) and 55- to 60-kDa C-terminal fragments, but the uncleaved/full-length form can also be isolated from brain extracts. As Slit2-N and full-length Slit2 bind tightly to cell membranes, we decided to explore the response of rat dorsal root ganglia (DRG) axons to substrate-bound Slit2 fragments in the stripe assay. Slit2 fragments were avoided by DRG axons when expressed on membranes or coated as stripes on laminin. However, when the Slit2 stripes were coated on fibronectin, DRG axons still avoided full-length Slit2 but grew preferentially on Slit2-N. DRG axon response to Slit2 fragments could be modulated by cGMP and by a laminin-1 peptide. These results strongly support the idea that extracellular matrix proteins modulate the response of growth cones to chemotropic molecules by modulating cyclic nucleotide levels. PMID- 11414794 TI - Epileptogenesis and enhanced prepulse inhibition in GABA(B1)-deficient mice. AB - The recent cloning of two GABA(B) receptor subunits, GABA(B1) and GABA(B2), has raised the possibility that differences in GABA(B) receptor subunit composition may give rise to pharmacologically or functionally distinct receptors. If present, such molecular diversity could permit the selective targeting of GABA(B) receptor subtypes specifically involved in pathologies such as drug addiction, spasticity, pain, and epilepsy. To address these issues we have developed a GABA(B1) subunit knockout mouse using gene targeting techniques. In the brains of GABA(B1) null mice, all pre- and postsynaptic GABA(B) receptor function was absent demonstrating that the GABA(B1) subunit is essential for all GABA(B) receptor-mediated mechanisms. Despite this, GABA(B1) null mice appeared normal at birth, although by postnatal week four their growth was retarded and they developed a generalized epilepsy that resulted in premature death. In addition, GABA(B1) heterozygote animals showed enhanced prepulse inhibition responses compared to littermate controls, suggesting that GABA(B1) deficient mice exhibit increased sensorimotor gating mechanisms. These data suggest that GABA(B) receptor antagonists may be of benefit in the treatment of psychiatric and neurological disorders in which attentional processing is impaired. PMID- 11414795 TI - Selective blockade of type-1 metabotropic glutamate receptors induces neuroprotection by enhancing gabaergic transmission. AB - Selective antagonists of mGlu1 (LY367385 and CPCCOEt) and mGlu5 (MPEP) metabotropic glutamate receptors were neuroprotective against NMDA toxicity when either applied to mixed cortical cultures or locally infused into the caudate nucleus. Neuroprotection produced by LY367385 or CPCCOEt was occluded by GABA and was abolished by a cocktail of GABA(A) and GABA(B) receptor antagonists. In contrast, GABAergic drugs did not influence the action of MPEP. In microdialysis studies, LY367385 and CPCCOEt substantially enhanced GABA release in the corpus striatum of freely moving animals, whereas MPEP had no effect on GABA but abolished the stimulation of glutamate release induced by NMDA. A role for mGlu1 receptors in modulating GABAergic transmission was supported by electrophysiological studies carried out in cortico-striatal slices. In this particular model, the mixed mGlu1/5 receptor agonist, DHPG, reduced bicuculline sensitive inhibitory postsynaptic currents presumably via a presynaptic mechanism. The action of DHPG was antagonized by LY367385, but not by MPEP. Taken together, these results indicate that selective blockade of mGlu1 receptors produces neuroprotection by enhancing GABAergic transmission. PMID- 11414796 TI - N-cadherin is involved in axon-oligodendrocyte contact and myelination. AB - We have analyzed the influence of the calcium-dependent cell adhesion molecule, N cadherin, on events leading to CNS myelination. Interactions between axons and oligodendrocyte progenitor (OP) cells and the CG4 OP cell line were examined by video-microscopy. OPs cocultured with dorsal root ganglia explants migrated around the culture and formed numerous contacts with axons. The duration of these contacts depended on the morphology of the OP, with OPs containing four or more processes forming long-lasting contacts and OPs with three or fewer processes forming short-termed contacts. Treatment with N-cadherin function blocking peptides approximately halved the duration of contacts made by cells with four or more processes but contact times for cells with three or less processes were unaffected. The L7 cadherin-blocking antibody and calcium withdrawal had similar effects. Contacts with axons regenerating from explants of adult retina, which do not have N-cadherin on their surface was examined. The contact duration of OPs to adult retina axons was short and similar in length to those formed between OPs and dorsal root ganglion axons in the presence of cadherin blocking reagents. Oligodendrocyte myelination was examined in organotypic rat cerebellar slice cultures, taken before myelination at postnatal day 10 and then allowed to myelinate in vitro over 7 days. When incubated with an N-cadherin function blocking peptide, myelination of Purkinje cell axons was reduced to about half of control levels, while control peptides were without effect. Cadherin-blockade did not prevent maturation of OPs, since oligodendrocytes showing myelin basic protein immunostaining were still found in these cultures. However, many of the cell processes did not colocalize with calbindin-positive axons. From these data we conclude that N-cadherin is important for the initial contact between a myelinating oligodendrocyte and axons and significantly contributes to the success of myelination. PMID- 11414797 TI - Bone marrow transfer from wild-type mice reverts the beneficial effect of genetically mediated immune deficiency in myelin mutants. AB - Inherited demyelinating neuropathies are chronically disabling human disorders caused by various genetic defects, including deletions, single site mutations, and duplications in the respective myelin genes. We have shown in a mouse model of one distinct hereditary demyelinating neuropathy (heterozygous P0-deficiency, P0+-) that an additional null mutation in the recombination activating gene-1 (RAG-1--) leads to a substantially milder disorder, indicating a disease modifying role of T-lymphocytes. In the present study, we addressed the role of lymphocytes in the mouse model by reconstituting bone marrow of P0+-/RAG-1-- mice with bone marrow from immunocompetent wild-type mice. We compared the pathology and nerve conduction in double mutant mice (P0+-/RAG-1-- on a C57BL/6 background) with that in double mutants after receiving a bone marrow transplant. We found that the milder demyelination seen in the lymphocyte-deficient P0+-/RAG-1-- mutants was reverted to the more severe pathology by reestablishing a competent immune system by bone marrow transfer. These data corroborate the concept that the immune system contributes substantially to the pathologic process in this mouse model and may open new avenues to ameliorate human hereditary neuropathies by exploiting immunosuppressive treatments. PMID- 11414798 TI - Intrahippocampal administration of an antibody against the HNK-1 carbohydrate impairs memory consolidation in an inhibitory learning task in mice. AB - Many cell adhesion molecules express the HNK-1 carbohydrate involved in formation and functioning of synapses. To assess its role in learning, we injected the monoclonal HNK-1 antibody or nonimmune IgG into the hippocampus of C57BL/6J mice 1 h after training in a step-down avoidance task. In animals treated with the HNK 1 antibody, latencies of step down in a recall session 48 h after injection did not change compared to training values and were significantly shorter versus IgG treated controls, which acquired the task normally. Similar differences between the two treatments were also observed after a stronger training protocol in a step-down avoidance paradigm. The HNK-1 antibody was effective only when injected 1 h, but not 48 h after training, thus affecting memory consolidation but not memory recall itself. The HNK-1 antibody impaired memory also in tenascin-R knock out mice, indicating that extracellular matrix molecule tenascin-R, one of the carriers of the HNK-1epitope in the hippocampus, does not mediate the function of the HNK-1 carbohydrate in this task. Our observations show that the HNK-1 carbohydrate is critically involved in memory consolidation in hippocampus dependent learning in mammals. PMID- 11414799 TI - Immunization for long-term protection against AIDS using the macaque model. PMID- 11414800 TI - A novel mechanism to ensure terminal initiation by hepatitis C virus NS5B polymerase. AB - Hepatitis C virus (HCV) nonstructural protein 5B (NS5B) RNA-dependent RNA polymerase (RdRp) has acquired a unique beta-hairpin in the thumb subdomain which protrudes toward the active site. We report here that this beta-hairpin plays an important role in positioning the 3' terminus of the viral RNA genome for correct initiation of replication. The presence of this beta-hairpin interferes with polymerase binding to preannealed double-stranded RNA (dsRNA) molecules and allows only the single-stranded 3' terminus of an RNA template to bind productively to the active site. We propose that this beta-hairpin may serve as a "gate" which prevents the 3' terminus of the template RNA from slipping through the active site and ensures initiation of replication from the terminus of the genome. This hypothesis is supported by the ability of a beta-hairpin deletion mutant that utilizes dsRNA substrates and initiates RNA synthesis internally. The proposed terminal initiation mechanism may represent a novel replication strategy adopted by HCV and related viruses. PMID- 11414801 TI - Induction of adult-like antibody, Th1, and CTL responses to measles hemagglutinin by early life murine immunization with an attenuated vaccinia-derived NYVAC(K1L) viral vector. AB - Although initially developed in adult animals, novel viral vectors expressing recombinant measles antigens must eventually prove their success in the early life setting, where the efficacy of the currently used live-attenuated measles virus vaccine is limited. The immunological requirements for vaccine candidates include the generation of protective antibody responses as well as the induction of Th1 and cytotoxic T lymphocytes (CTL) responses, which is challenging in the neonatal setting. Here, we report that young BALB/c mice immunized with a single dose of a vaccinia-based NYVAC(K1L) vector generate adult-like antihemagglutinin (HA) antibody responses as well as adult-like Th1 and CTL responses. Despite this strong immunogenicity in early life, antibody responses (but not T-cell responses) to a single dose of NYVAC(K1L)-HA remained susceptible to inhibition by preexisting measles antibodies, calling for use of prime-boost strategies. NYVAC(K1L)-HA is the first attenuated live viral vector demonstrated as capable of inducing adult-like antibody, Th1, and CTL responses against measles in an early life murine immunization model, a capacity previously only reported for measles DNA vaccines. PMID- 11414802 TI - Mouse hepatitis virus replicase protein complexes are translocated to sites of M protein accumulation in the ERGIC at late times of infection. AB - The coronavirus mouse hepatitis virus (MHV) directs the synthesis of viral RNA on discrete membranous complexes that are distributed throughout the cell cytoplasm. These putative replication complexes are composed of intimately associated but biochemically distinct membrane populations, each of which contains proteins processed from the replicase (gene 1) polyprotein. Specifically, one membrane population contains the gene 1 proteins p65 and p1a-22, while the other contains the gene 1 proteins p28 and helicase, as well as the structural nucleocapsid (N) protein and newly synthesized viral RNA. In this study, immunofluorescence confocal microscopy was used to define the relationship of the membrane populations comprising the putative replication complexes at different times of infection in MHV-A59-infected delayed brain tumor cells. At 5.5 h postinfection (p.i.) the membranes containing N and helicase colocalized with the membranes containing p1a-22/p65 at foci distinct from sites of M accumulation. By 8 to 12 h p.i., however, the membranes containing helicase and N had a predominantly perinuclear distribution and colocalized with M. In contrast, the p1a-22/p65 containing membranes retained a peripheral, punctate distribution at all times of infection and did not colocalize with M. By late times of infection, helicase, N, and M each also colocalized with ERGIC p53, a specific marker for the endoplasmic reticulum-Golgi-intermediate compartment. These data demonstrated that the putative replication complexes separated into component membranes that relocalized during the course of infection. These results suggest that the membrane populations within the MHV replication complex serve distinct functions both in RNA synthesis and in delivery of replication products to sites of virus assembly. PMID- 11414803 TI - Binding of PKA-RIIalpha to the Adenovirus E1A12S oncoprotein correlates with its nuclear translocation and an increase in PKA-dependent promoter activity. AB - The adenovirus type 12 (Ad12) E1A12S oncoprotein utilizes the cAMP/protein kinase A (PKA) signal transduction pathway to activate expression of the viral E2 gene, the products of which are essential for viral replication. A central unsolved question is, however, whether E1A12S interacts directly with PKA in the process of promoter activation. We show here that E1A12S binds to the regulatory subunits (R) of PKA in vitro and in vivo. Interaction depends on the N-terminus and the conserved region 1 (CR1) of E1A12S. Both domains are also essential for the activation of viral E2 gene expression. Infection of cells with Ad12 leads to the cellular redistribution of RIIalpha from the cytoplasm into the nucleus. Furthermore, RIIalpha is also located in the nucleus of cells transformed by E1 of Ad12 and transient expression of E1A12S leads to the redistribution of RIIalpha into the nucleus in a N-terminus- and CR1-dependent manner. Cotransfection of E1A12S with RIIalpha results in strong activation of the E2 promoter. Based on these results we conclude that E1A12S functions as a viral A kinase anchoring protein redistributing RIIalpha from the cytoplasm into the nucleus where it is involved in E1A12S-mediated activation of the E2 promoter. PMID- 11414804 TI - VZV gB endocytosis and Golgi localization are mediated by YXXphi motifs in its cytoplasmic domain. AB - The cytoplasmic domains of many membrane proteins contain sorting signals that mediate their endocytosis from the plasma membrane. VZV gB contains three consensus internalization motifs within its cytoplasmic domain: YMTL (aa 818 821), YSRV (aa 857-860), and LL (aa 841-842). To determine whether VZV gB is internalized from the plasma membrane, and whether these motifs are required for its endocytosis, we compared the internalization of native gB to that of gB containing mutations in each of the predicted internalization motifs. VZV gB present on the surface of transfected cells associated with clathrin and was efficiently internalized to the Golgi apparatus within 60 min at 37 degrees C. VZV gB containing the mutation Y857 failed to be internalized, while gB-Y818A was internalized but did not accumulate in the Golgi. These data indicate that the internalization of VZV gB, and its subsequent localization to the Golgi, is mediated by two tyrosine-based sequence motifs in its cytoplasmic domain. PMID- 11414805 TI - Induction of apoptosis in vitro by the 17-kDa nonstructural protein of infectious bursal disease virus: possible role in viral pathogenesis. AB - Infectious bursal disease virus (IBDV) causes severe immunodeficiency in young chickens by destroying the precursors of antibody-producing B cells in the bursa of Fabricius. It has been shown that IBDV infection induces apoptosis in chicken embryo and tissue culture cells. We previously reported that an IBDV mutant lacking the expression of 17-kDa nonstructural (NS) protein exhibited decreased apoptotic effects in cell culture as compared to the parental IBDV, suggesting that the NS protein may be involved in induction of apoptosis. Here, we report that the NS protein of IBDV alone is capable of inducing apoptosis in cell culture. Transfection of chicken B-lymphocyte cell line (RP9) and chicken embryo fibroblast cells with a plasmid DNA, containing the NS protein gene under the control of the immediate-early promoter-enhancer region of human cytomegalovirus, induced programmed cell death in both cell lines. Apoptosis changes, such as chromatin condensation, DNA fragmentation, and the appearance of apoptotic nuclear bodies, were observed in cell cultures 48-h posttransfection. As reported earlier, the mutant IBDV grew to lower titers with slower replication kinetics and lower cytopathogenicity when compared to that of the parental virus. Here, we demonstrate that the mutant virus is closely associated with cells and its yield from the supernatant was approximately 30-fold lower than the wild-type due to increased cell association, indicating a deficiency in lysis of virus-infected cells. Taken together, our results indicate that the NS protein of IBDV is highly cytotoxic, which brings about the release of the viral progeny from cells, and thus play an important role in viral pathogenesis. PMID- 11414806 TI - Plants expressing tomato golden mosaic virus AL2 or beet curly top virus L2 transgenes show enhanced susceptibility to infection by DNA and RNA viruses. AB - The AL2 gene of the geminivirus tomato golden mosaic virus (TGMV) encodes a transcriptional activator protein (TrAP) that is required for efficient expression of the viral coat protein (CP) and BR1 gene promoters. In contrast, L2, the positional homolog of AL2 in the related beet curly top virus (BCTV), is not required for CP expression, raising questions about the functional relationship between the AL2 and L2 gene products. In this study, transgenic Nicotiana benthamiana and N. tabacum var. Samsun plants expressing a truncated AL2 gene (AL2(1-100), lacking the activation domain) or full-length L2 were prepared. These transgenic plants showed a novel enhanced susceptibility (ES) phenotype following inoculation with TGMV, BCTV, or tobacco mosaic virus (TMV), an unrelated RNA virus. ES is characterized by a reduction in the mean latent period (from 1 to 9 days) and by a decrease in the inoculum concentration required to infect transgenic plants (ID50 reduced 6- to 60-fold). However, ES does not result in an enhancement of disease symptoms, and viral nucleic acids do not accumulate to substantially greater levels in infected transgenic plants. That both viral transgenes condition ES suggests that their products share the ability to suppress a host stress or defense response that acts against DNA and RNA viruses. The data further indicate that the transcriptional activation activity of AL2 protein is not required for suppression. The nature of the response targeted by the AL2 and L2 gene products is discussed. PMID- 11414807 TI - Long-distance movement and replication maintenance functions correlate with silencing suppression activity of potyviral HC-Pro. AB - The tobacco etch potyviral protein, HC-Pro, is a multifunctional proteinase required for long-distance movement in plants and maintenance of genome replication at the single-cell level. It also functions in a counterdefensive capacity as a suppressor of posttranscriptional gene silencing (PTGS). To determine whether the requirements for HC-Pro during long distance movement and replication maintenance are due to the silencing suppressor function of the protein, a series of HC-Pro alanine scanning and other site-directed mutants were analyzed. Using a transient silencing suppression assay in Agrobacterium-injected leaf tissue, several suppression-defective mutants were identified. Each of six HC-Pro mutations, which were shown previously to confer long-distance movement and replication maintenance defects, conferred PTGS suppression defects. Interestingly, the genes encoding these defective HC-Pro derivatives were themselves susceptible targets of PTGS, resulting in low levels of mRNA and protein accumulation. Mutations that inactivated the proteinase domain active site had no effect on PTGS suppression function. The results are consistent with the hypothesis that the role of HC-Pro in long-distance movement and genome replication depends on PTGS suppression function and that this function is independent of HC-Pro proteolytic activity. PMID- 11414808 TI - Differential effects of parainfluenza virus type 3 on human monocytes and dendritic cells. AB - To understand the lack of protective immunity observed after infection with parainfluenza virus type 3 (PIV3), we tested the effect of the virus on human monocytes and monocyte-derived immature dendritic cells (DCs). Expression of viral antigens on the cell surfaces correlated with replication of the virus, which was marginal in monocytes but extremely efficient in DCs. The virus increased monocyte survival at least in part through the production of granulocyte-macrophage colony-stimulating factor but, in contrast, accelerated DC apoptosis. In addition, PIV3 infection failed to activate monocytes but induced maturation of DCs with increased expression of CD54, HLA-DR, CD86, and CD83 and production of bioactive IL-12. However, PIV3-infected DCs demonstrated low stimulatory properties in DC-T cell cocultures, a finding that could not be attributed to the production of infectious virus or IL-10. These results demonstrate for the first time that PIV3 dramatically modifies the survival and/or the function of antigen-presenting cells and might therefore prevent the development of efficient antiviral immune responses. PMID- 11414809 TI - Nuclear localization of the ORF2 protein encoded by porcine circovirus type 2. AB - Infectious porcine circovirus type 2 (PCV2) was generated following transfection of a porcine retina cell line (VIDO R1) with cloned circovirus DNA. Expression of open reading frame 2 (ORF2) was detected at 24 h postinfection and onwards increasingly throughout the infection by Western blot analysis using ORF2 specific polyclonal antibody. Moreover, the ORF2 protein was also detected in purified PCV2 virus, indicating that ORF2 is a structural component of PCV2 viral capsid. Nuclear localization of PCV2 ORF2 was demonstrated by immunofluorescence assay in PCV2-infected cells. An analysis of the subcellular localization of a series of truncation mutants of ORF2 fused with the green fluorescent protein indicated that the nuclear localization signal of ORF2 was conferred by the N terminal 41 amino acids. This domain was further analyzed through site-directed mutagenesis, suggesting that the presence of basic amino acid residues at positions 12 to 18 and 34 to 41 are important for the strict nuclear targeting of PCV2 ORF2. PMID- 11414810 TI - Mutations in the measles virus C protein that up regulate viral RNA synthesis. AB - The measles virus RNA-dependent RNA polymerase consists of two virus-encoded subunits, the phosphoprotein (P) and the large (L) protein. The P mRNA also codes for a C protein in the +1 reading frame relative to P. The activities of the measles P and C proteins from the vaccine strain, EdB, a wild-type CM strain, and an SSPE P4 strain were investigated using a CAT reporter minigenome assay. CAT is synthesized following replication and transcription of a DI-CAT minigenome supported by individual P, L, and N plasmids expressed in a mammalian expression system. As measured by CAT activity, CMP1 and P4P1 stimulate transcription and replication four- to six- and six- to eightfold, respectively, better than EdP. There are 10 and 16 amino acid changes in the P protein and three and four changes in C in CMP1 and P4P1, respectively, relative to EdP. By constructing chimeric P genes we showed that mutations throughout P4P1 were required for enhanced polymerase activity, while only mutations in the 5'-terminal portion, encompassing the C ORF, of the CMP1 gene mediated stimulation. Abrogation of C expression from the Ed and CM P genes resulted in an increase in RNA synthesis of twofold for CMP1S and four- to fivefold for EdPS. With the addition of C protein expressed from a separate plasmid that contains only the C ORF, EdC reduces viral RNA synthesis more strongly than CMC. These data suggest that EdC and CMC proteins give a differential inhibition that accounts for most of the differences in RNA synthesis by EdP and CMP1. PMID- 11414811 TI - Extreme genetic diversity among Pirital virus (Arenaviridae) isolates from western Venezuela. AB - Pirital-like virus isolates from rodents collected in a variety of habitats within a six-state area of central Venezuela were analyzed genetically by amplifying a portion of the nucleocapsid protein gene using RT-PCR. Comparisons of the sequences from 30 selected Pirital-like virus isolates demonstrated up to 26% divergence in nucleotide sequences and up to 16% divergence in deduced amino acid sequences. Within the Pirital monophyletic group, 14 distinct lineages or genotypes, differing by at least 6% in nucleotide sequences, were identified. Although sample sizes were small for some lineages, many of the different genotypes were sampled in only one region or locality, suggesting allopatric divergence. Complement fixation tests with representatives of the most divergent Pirital virus lineages failed to delineate multiple species or subtypes within the Pirital clade. These results indicate that the previously proposed 12% nucleocapsid protein amino acid sequence divergence cutoff value for delineating arenavirus species is not appropriate for the entire family. When individual clones were examined from PCR amplicons, a mean of 0.17% sequence diversity vs the consensus sequences was detected, suggesting diverse quasispecies populations within infected rodent hosts. Possible explanations for the extreme genetic diversity within and among Pirital virus populations in infected rodents are discussed. PMID- 11414812 TI - TATA box occupancy in the SV40 transcription elongation complex. AB - In order to gain insight into requirements for template activation and commitment in mammalian transcription, TATA site occupancy was measured in native SV40 viral transcription complexes that were in the process of transcription elongation at the time of cell lysis. This was accomplished by quantifying resistance to restriction enzyme digestion of transcription complexes in nuclear lysate. The rate of cleavage at the TATA site of the late gene in the native complex was slower than that of a bare DNA control, both for wild-type virus and for a virus containing a TATA consensus sequence. These results suggest that the TATA site in the transcription elongation complex in vivo is occupied with transcription factor TBP/TFIID. When considered in light of previous work, these findings support a model in which transcription activation involves reinitiation from a promoter that contains both activator and TFIID bound in a stable complex. PMID- 11414813 TI - HIV envelope proteins differentially utilize CXCR4 and CCR5 coreceptors for induction of apoptosis. AB - The involvement of CXCR4 and CCR5 coreceptors in apoptosis induced by the HIV envelope (Env) proteins has not been well defined. We found that simian human immunodeficiency virus (SHIV) virus-like particles (VLPs) containing HIV Env proteins preferentially induce apoptosis of cells corresponding to their coreceptor usage in a CD4+ T cell line. We also demonstrated that induction of apoptosis by SHIV VLPs is correlated with coreceptor usage in a non-T cell line. We examined the effects of SHIV VLPs containing Env proteins derived from either a T-cell-tropic HIV (BH10) strain or a dual-tropic HIV (89.6) strain on induction of apoptosis in recombinant CD4+ human osteosarcoma (HOS) cells expressing either CXCR4 (HOS-CD4.CXCR4) or CCR5 coreceptors (HOS-CD4.CCR5). HOS-CD4.CXCR4 or HOS CD4.CCR5 cells were activated with concanavalin A and cocultured with VLPs. By TUNEL (TdT-mediated dUTP-X nick end labeling) fluorescence staining and flow cytometry assays, SHIV BH10 VLPs were found to preferentially induce apoptosis in HOS-CD4.CXCR4 cells but not in HOS-CD4 or HOS-CD4.CCR5 cells. On the other hand, SHIV 89.6 VLPs induced an elevated level of apoptosis in both HOS-CD4.CXCR4 and HOS-CD4.CCR5 cells in a dose-dependent fashion. These data demonstrate that T cell-tropic BH10 Env preferentially utilizes CXCR4, but not CCR5, for induction of apoptosis, whereas dual-tropic 89.6 Env induces apoptosis in both CXCR4- and CCR5-containing cell lines. PMID- 11414814 TI - The HDV large-delta antigen fused with GFP remains functional and provides for studying its dynamic distribution. AB - Hepatitis D virus (HDV) requires the isoprenylated large delta antigen (LDAg) for interaction with hepatitis B surface antigen (HBsAg) to allow packaging and secretion out of the host cell. Phosphorylated LDAg has been found but, as yet, neither localization of LDAg within the nucleus nor any other function has been correlated with modification. In this study, we transfected HuH-7 or HeLa cells with plasmids encoding various lengths of LDAg [designated GFP-LD and GFP-LD(31 214) for full length and a deletion, respectively] or non-isoprenylated mutants of these [designated GFP-LDM and GFP-LD(31-214)M] fused to the green fluorescent protein (GFP). These fusion proteins were then characterized and it was found that: (i) the addition of the GFP did not interfere with the functioning of the full-length or N-terminally deleted LDAgs when interacting with HBsAg for secretion; (ii) the HDV small antigen (SDAg) together with the GFP-LD, but not the GFP-LD(31-214), could be cosecreted by HBsAg; and (iii) the GFP-LD, but not the GFP-LD(31-214), exerted a dominant-negative role on HDV genome replication. Analyses of transiently transfected cells and postmitotic permanent cells revealed the sequential appearance of GFP-LD in the nucleoplasm, then in the nucleolus, and finally in nuclear speckles (NS). Isoprenylation of LDAg seems to be important for targeting to and accumulating in the NS, which was evident from the dynamic and static localization of the non-isoprenylation mutant (GFP-LDM) and the distribution of wild-type (GFP-LD) when treated with an isoprenylation inhibitor, lovastatin, for more than 48 h. Permanently expressing GFP-LD cells allowed us to show the dynamic redistribution of dephosphorylated GFP-LD from the nucleolus to the SC-35 containing NS in the presence of dichlororibofuranosyl benzimidazole (DRB) and then the translocation back of the GFP-LD to the nucleolus within 2 h after removal of DRB. Our studies thus suggest that the various versions of the GFP-LD fusion protein, having the same function as their nonfusion counterparts, can be a powerful tool for the study of the dynamic localization of LDAg when correlated with the functional modification of this protein. PMID- 11414815 TI - Role of individual oligosaccharide chains in antigenic properties, intracellular transport, and biological activities of influenza C virus hemagglutinin-esterase protein. AB - The hemagglutinin-esterase (HE) glycoprotein of influenza C virus is composed of three domains: a stem domain active in membrane fusion (F), an acetylesterase domain (E), and a receptor-binding domain (R). The protein contains eight N linked glycosylation sites, four (positions 26, 395, 552, and 603) in the F domain, three (positions 61, 131, and 144) in the E domain, and one (position 189) in the R domain. Here, we investigated the role of the individual oligosaccharide chains in antigenic properties, intracellular transport, and biological activities of the HE protein by eliminating each of the glycosylation sites by site-specific mutagenesis. Comparison of electrophoretic mobility between the wild-type and the mutant proteins showed that while seven of the glycosylation sites are used, one (position 131) is not. Analysis of reactivity of the mutants with anti-HE monoclonal antibodies demonstrated that glycosylation at position 144 is essential for the formation of conformation-dependent epitopes. It was also evident that glycosylation at the two sites in the F domain (positions 26 and 603), in addition to that in the E domain (position 144), is required for the HE molecule to be transported from the endoplasmic reticulum and that mutant HEs lacking one of these three sites failed to undergo the trimer assembly. Removal of an oligosaccharide chain at position 144 or 189 resulted in a decrease in the esterase activity. By contrast, two mutants lacking an oligosaccharide chain at position 26 or 603, which were defective not only in cell surface expression but in trimerization, possessed full-enzyme activity, suggesting that the HE monomers present within the cell have acetylesterase activity. Fusion activity of cells expressing each of mutant HEs was found to be comparable with the ability of the protein to be transported to the cell surface, suggesting that there is no specific oligosaccharide chain that plays a critical role in promoting membrane fusion. PMID- 11414816 TI - Specific packaging of nodaviral RNA2 requires the N-terminus of the capsid protein. AB - Flock house virus (FHV), a member of the family Nodaviridae, is a nonenveloped, icosahedral insect virus whose capsids are assembled from 180 copies of a single type of coat protein. The viral genome is split between two segments of single stranded positive-sense RNA, RNA1 and RNA2, which are packaged into a single virion. We previously demonstrated that synthesis of FHV coat protein in the baculovirus expression system results in assembly of virus-like particles whose capsids are indistinguishable from those of native virions, although the encapsidated RNA represents primarily cellular RNA. In contrast, expression of a deletion mutant lacking N-terminal residues 2-31 results in formation of multiple types of particles which differ in size, shape, and RNA contents. We postulated that the polymorphism was imposed by the type of RNA that the coat protein selected for packaging. In the current study we tested this hypothesis by analyzing the assembly of the mutant coat protein in Drosophila cells in the presence of replicating FHV RNAs. As anticipated, the resulting particles had the same shape and dimensions as wt virions. Surprisingly, however, they contained little RNA2 while packaging of RNA1 was not affected. Small amounts of defective interfering RNAs, which emerged rapidly in the presence of the mutant coat protein, were also detected. Taken together, these observations confirm our earlier hypothesis that selection of nonviral RNAs for packaging can significantly alter the assembly process. In addition, they demonstrate that the N-terminus of the FHV coat protein contains important determinants for recognition and packaging of RNA2. Our results provide the first evidence that encapsidation of the two genomic RNAs occurs independently and that the coat protein uses different regions for the recognition of RNA1 and RNA2. PMID- 11414817 TI - Cryoreduction of methyl-coenzyme M reductase: EPR characterization of forms, MCR(ox1) and MCR (red1). AB - Methyl-coenzyme M reductase (MCR) catalyzes the formation of methyl-coenzyme M (CH(3)S-CH(2)CH(2)SO(3)) from methane. The active site is a nickel tetrahydrocorphinoid cofactor, factor 430, which in inactive form contains EPR silent Ni(II). Two such forms, denoted MCR(silent) and MCR(ox1)(-)(silent), were previously structurally characterized by X-ray crystallography. We describe here the cryoreduction of both of these MCR forms by gamma-irradiation at 77 K, which yields reduced protein maintaining the structure of the oxidized starting material. Cryoreduction of MCR(silent) yields an EPR signal that strongly resembles that of MCR(red1), the active form of MCR; and stepwise annealing to 260-270 K leads to formation of MCR(red1). Cryoreduction of MCR(ox1)(-)(silent) solutions shows that our preparative method for this state yields enzyme that contains two major forms. One behaves similarly to MCR(silent), as shown by the observation that both of these forms give essentially the same redlike EPR signals upon cryoreduction, both of which give MCR(red1) upon annealing. The other form is assigned to the crystallographically characterized MCR(ox1)( )(silent) and directly gives MCR(ox1) upon cryoreduction. X-band spectra of these cryoreduced samples, and of conventionally prepared MCR(red1) and MCR(ox1), all show resolved hyperfine splitting from four equivalent nitrogen ligands with coupling constants in agreement with those determined in previous EPR studies and from (14)N ENDOR of MCR(red1) and MCR(ox1). These experiments have confirmed that all EPR-visible forms of MCR contain Ni(I) and for the first time generated in vitro the EPR-visible, enzymatically active MCR(red1) and the activate-able "ready" MCR(ox1) from "silent" precursors. Because the solution Ni(II) species we assign as MCR(ox1)(-)(silent) gives as its primary cryoreduction product the Ni(I) state MCR(ox1), previous crystallographic data on MCR(ox1)(-)(silent) allow us to identify the exogenous axial ligand in MCR(ox1) as the thiolate from CoM; the cryoreduction experiments further allow us to propose possible axial ligands in MCR(red1). The availability of model compounds for MCR(red1) and MCR(ox1) also is discussed. PMID- 11414818 TI - Bicarbonate as a proton donor in catalysis by Zn(II)- and Co(II)-containing carbonic anhydrases. AB - Catalysis of (18)O exchange between CO(2) and water catalyzed by a Co(II) substituted mutant of human carbonic anhydrase II is analyzed to show the rate of release of H(2)(18)O from the active site. This rate, measured by mass spectrometry, is dependent on proton transfer to the metal-bound (18)O-labeled hydroxide, and was observed in a site-specific mutant of carbonic anhydrase II in which a prominent proton shuttle residue His64 was replaced by alanine, which does not support proton transport. Upon increasing the concentration of bicarbonate, the rate of release of H(2)(18)O increased in a saturable manner to a maximum of 4 x 10(5) s(-)(1), consistent with proton transfer from bicarbonate to the Co(II)-bound hydroxide. The same mutant of carbonic anhydrase containing Zn(II) had the rate of release of H(2)(18)O smaller by 10-fold, but rate of interconversion of CO(2) and HCO(3)(-) about the same as the Co(II)-containing enzyme. These data as well as solvent hydrogen isotope effects suggest that the bicarbonate transferring the proton is bound to the cobalt in the enzyme. The enhancement of (18)O exchange caused by increasing bicarbonate concentration during catalysis by the Zn(II)-containing carbonic anhydrase from the archaeon Methanosarcina thermophila suggests that a very similar mechanism for proton donation by bicarbonate occurs with this wild-type enzyme. PMID- 11414819 TI - Guanine oxidation: NMR characterization of a dehydro-guanidinohydantoin residue generated by a 2e-oxidation of d(GpT). AB - The Mn-TMPyP/KHSO(5) system was used to oxidize guanine contained within a dinucleoside monophosphate d(GpT). To identify the guanine oxidation product having a mass with 4 amu above the mass of guanine itself, this relatively unstable compound was reduced to a more stable one. The ESI/MS and NMR data allowed us to propose a dehydro-guanidinohydantoin structure for the (G+4) guanine oxidation product. PMID- 11414820 TI - A simple, high-resolution method for establishing DNA binding affinity and sequence selectivity. AB - Full details of the development of a simple, nondestructive, and high-throughput method for establishing DNA binding affinity and sequence selectivity are described. The method is based on the loss of fluorescence derived from the displacement of ethidium bromide or thiazole orange from the DNA of interest or, in selected instances, the change in intrinsic fluorescence of a DNA binding agent itself and is applicable for assessing relative or absolute DNA binding affinities. Enlisting a library of hairpin deoxyoligonucleotides containing all five base pair (512 hairpins) or four base pair (136 hairpins) sequences displayed in a 96-well format, a compound's rank order binding to all possible sequences is generated, resulting in a high-resolution definition of its sequence selectivity using this fluorescent intercalator displacement (FID) assay. As such, the technique complements the use of footprinting or affinity cleavage for the establishment of DNA binding selectivity and provides the information at a higher resolution. The merged bar graphs generated by this rank order binding provide a qualitative way to compare, or profile, DNA binding affinity and selectivity. The 96-well format assay (512 hairpins) can be conducted at a minimal cost (presently ca. $100 for hairpin deoxyoligonucleotides/assay with ethiduim bromide or less with thiazole orange), with a rapid readout using a fluorescent plate reader (15 min), and is adaptable to automation (Tecan Genesis Workstation 100 robotic system). Its use in generating a profile of DNA binding selectivity for several agents including distamycin A, netropsin, DAPI, Hoechst 33258, and berenil is described. Techniques for establishing binding constants from quantitative titrations are compared, and recommendations are made for use of a Scatchard or curve fitting analysis of the titration binding curves as a reliable means to quantitate the binding affinity. PMID- 11414821 TI - Synthesis of ent-alantrypinone. AB - This paper presents a synthesis of ent-alantrypinone (ent-6), the enantiomer of a natural product produced by the fungus Penicillium thymicola. The synthesis revolves around the Li[Me(3)AlSPh]-promoted isomerization of iminobenzoxazine 33 to quinazolinone 34, an N-acyliminium ion cyclization that converts enamide 9 to bridged indole 35, and rearrangement of 35 to oxindole ent-6. Ancillary chemistry that involves thermal fragmentation of an iminobenzoxazine to a nitrile ylide and Me(2)AlSPh-mediated cyclization of oxime ether-ester 22 to pyrrolidinone 23 is also described. PMID- 11414822 TI - Proton transfer reactions of methylanthracene radical cations with pyridine bases under non-steady-state conditions. Real kinetic isotope effect evidence for extensive tunneling. AB - The kinetics of the proton transfer reactions between the 9-methyl-10 phenylanthracene radical cation (MPA(+)(.)) with 2,6-lutidine were studied in acetonitrile-Bu(4)NBF(4) (0.1 M) using derivative cyclic voltammetry. Comparisons of extent of reaction-time profiles with theoretical data for both the simple single-step proton transfer and a mechanism involving the formation of a donor acceptor complex prior to unimolecular proton transfer were made. The experimental extent of reaction-time profiles deviated significantly from those simulated for the single-step mechanism, while excellent fits of experimental to theoretical data, in the pre-steady-state period, for the complex mechanism were observed. In this time period, the apparent deuterium kinetic isotope effects (KIE(app)) were observed to vary significantly with the extent of reaction as predicted by the complex mechanism. Resolution of the apparent rate constants into the microscopic rate constants for the complex mechanism resulted in a real kinetic isotope effect (KIE(real)) equal to 82 at 291 K. Arrhenius activation parameters (252-312 K) for the reactions of MPA(+)(*) with 2,6-lutidine in acetonitrile-Bu(4)NBF(4) (0.1 M) revealed E(a)(D) - E(a)(H) equal to 2.89 kcal/mol and A(D)/A(H) equal to 2.09. In this temperature range, KIE(real) varied from 46 at the highest temperature to 134 at the lowest. The large KIE(real), along with the Arrhenius parameters, are indicative of extensive tunneling for the proton transfer steps. PMID- 11414823 TI - Fast and convenient divergent synthesis of aliphatic ester dendrimers by anhydride coupling. AB - A novel divergent approach was developed for the synthesis of dendritic aliphatic polyester structures using an acetal-protected anhydride derivative of 2,2 bis(hydroxymethyl)propionic acid as the acylating agent. This divergent synthesis is remarkable, because unlike all others, it only requires a small excess of reagent to achieve quantitative growth, and it requires no means of purification other than a simple solvent extraction or precipitation. A monodisperse sixth generation dendrimer with molecular weight of 30 711 Dalton and 192 masked hydroxyl groups was prepared in high yield and purity using 1,1,1 tris(hydroxyphenyl)ethane as the core molecule. Linear and star-shaped poly(ethylene glycol) (PEG) derivatives of narrow polydispersity were also used as core molecules in the divergent synthesis of dendritic-linear copolymer hybrids up to the fourth generation without requiring any chromatographic purification. PMID- 11414824 TI - Applications of vinylogous Mannich reactions. Total syntheses of the Ergot alkaloids rugulovasines A and B and setoclavine. AB - Concise syntheses of the Ergot alkaloids rugulovasine A (3a), rugulovasine B (3b), and setoclavine (2) have been completed by strategies that feature inter- and intramolecular vinylogous Mannich reactions as the key steps. Thus, the first synthesis of 3a,b commenced with the conversion of the known indole 17 into 24 via the addition of the furan 22 to the iminium ion 21, which was generated in situ from the aldehyde 19. Cyclization of 24 by a novel S(RN)1 reaction followed by removal of the N-benzyl group furnished a mixture (1:2) of 3a and 3b. In an alternative approach to these alkaloids, the biaryl 35 was reduced with DIBAL-H to give an intermediate imine that underwent spontaneous cyclization via an intramolecular vinylogous Mannich addition to provide 36a,b. N-Methylation of the derived benzyl carbamates 37a,b followed by global deprotection gave a mixture (2:1) of rugulovasines A and B (3a,b). Setoclavine (2) was then prepared from the biaryl 41 using a closely related intramolecular vinylogous Mannich reaction to furnish the spirocyclic lactones 42a,b. These lactones were subsequently transformed by hydride reduction and reductive methylation into the ergoline derivatives 43a,b, which were in turn converted into 2 by deprotection and solvolytic 1,3-rearrangement of the allylic hydroxyl group. PMID- 11414825 TI - Total synthesis of (-)-cylindrocyclophanes A and F exploiting the reversible nature of the olefin cross metathesis reaction. AB - Efficient total syntheses of the C(2)-symmetric (-)-cylindrocyclophanes A and F (1a and 1f) have been achieved. The initial strategy featured the use of a common advanced intermediate to assemble in stepwise fashion the required macrocycle of 1f, exploiting in turn a Myers reductive coupling followed by ring-closing metathesis. In a second-generation strategy, a remarkable cross olefin metathesis dimerization cascade was discovered and exploited to assemble the requisite [7,7] paracyclophane macrocycles of both 1a and 1f from dienyl monomers. The successful syntheses also featured the effective use of the Danheiser annulation to construct substrates for both the Myers reductive coupling and the metathesis dimerizations strategies. Finally, the Kowalski two-step chain homologation of esters to siloxyalkynes proved superior over the original one-step protocol. PMID- 11414826 TI - Hydrogen bond breaking dynamics of the water trimer in the translational and librational band region of liquid water. AB - The effect of exciting each of the three classes of intermolecular vibrations on the hydrogen bond lifetime (tau(H)) of the isolated water trimer is investigated by far-infrared laser spectroscopy. Single excitation of a librational vibration decreases tau(H) by 3 orders of magnitude to tau(H) = 1-6 ps, comparable to the time scale of a number of important bulk water dynamical relaxation processes. In contrast, excitation of translational or torsional vibrations has no significant effect (tau(H) = 1-2 ns). Although such a dependence of tau(H) on intermolecular motions has also been proposed for liquid water via computer simulations, these are the first experiments that provide a detailed molecular picture of the respective motions without extensive interpretation. PMID- 11414827 TI - A nanometer-sized high-spin polyradical: poly(4-phenoxyl-1,2-phenylenevinylene) planarily extended in a non-Kekule fashion and its magnetic force microscopic images. AB - A pi-conjugated, but non-Kekule- and nondisjoint-type poly(1,2-phenylenevinylene) network bearing 4-substituted di-tert-butylphenoxyls was synthesized through a one-pot polycondensation of the star-shaped subpart and the subsequent oxidation, which was persistent even at room temperature. The polyphenoxyl radical with a spin concentration of 0.4 displayed an average S of 10/2. The polyradical with the molecular weight of 3.2 x 10(4) gave a disklike image of ca. 35 x 0.6 nm with both an atomic and a magnetic force microscopy: the molecular image was examined as a nanoscale and single-molecular-based magnetic dot. PMID- 11414828 TI - Internucleobase-interaction-directed self-assembly of nanofibers from homo- and heteroditopic 1,omega-nucleobase bolaamphiphiles. AB - The complementary 1,omega-thymine, 1,omega-adenine, and 1,omega-(thymine, adenine) bolaamphiphiles, [N,N'-bis[3-(2,4-dihydroxy-5-methylpyrimidine-1 yl)propionyl]1,n-diaminoalkane [T-n-T (n = 10, 11, 12)], N, N'-bis[3-(6 aminopurine-9-yl)propionyl]1,n-diaminoalkane [A-n-A (n = 10, 11, 12)], and N-[3 (2,4-dihydroxy-5-methylpyrimidine-1-yl)propionyl], N'-[3-(6-aminopurine-9 yl)propionyl]1,n-diaminoalkane [T-n-A (n = 10, 11, 12)], respectively] have been synthesized. The spontaneous homo- and heteroassembly of these nucleobase-based bolaamphiphiles has been studied by light microscopy, energy-filtering transmission electron microscopy, FT-IR, and powder X-ray diffraction analyses. The achiral T-10-T bolaamphiphile produced in 10% ethanolic/aqueous solutions unprecedented double-helical ropes of 1-2 microm in widths and several hundred micrometers in length, whereas the complementary homologue A-10-A gave only microcrystalline solids of 1-10 microm in size. In contrast, an equimolar mixture of T-10-T and A-10-A yielded supramolecular fibers of 15-30 nm in width. (1)H NMR, CD, and UV studies of solution photoreactions of T-10-T suggested that under natural light the chiral rope formation is triggered by photodimerization of trace amounts of the thymine moieties in the T-10-T assemblies. Complementary hydrogen bond formation between the thymine-adenine heterobase pairs was found to prevent such a photoreaction and resulted in no chiral rope formation. The heteroditopic T-12-A bolaamphiphile self-assembled to form supramolecular fibers. Multilamellar organization was proposed for the homo- and heteroassemblies made of T-n-T and A-n-A. PMID- 11414829 TI - Reactivity at the interface of chiral amphiphilic dendrimers. High asymmetric reduction by NaBH(4) of various prochiral ketones. AB - New amphiphilic dendrimers derived from PAMAM and D-gluconolactone were found to induce chirality in the reduction of prochiral ketones by NaBH(4), in heterogeneous (THF) and homogeneous (water) conditions. The third generation of these amphiphilic dendrimers, G(3)G, was found to be a good chiral ligand for the reduction of various prochiral ketones in heterogeneous conditions. Even with substrates well-known to give poor results (especially linear ketones), good enantioselectivities were obtained. It is also important to notice that under heterogeneous conditions (THF) the dendrimer could be recovered by filtration, regenerated, and recycled (up to 10 times), leading to reproducible results in asymmetric reduction of ketones. We have also discussed the reduction of acetophenone in water. Evidence is presented that the selectivity is dominated by the architecture of the dendrimer and some supramolecular ordering in the position of the ketone at the chiral solvating interface. The results obtained showed a correlation between stereoselectivity of the reduction and the compact character of the dendritic particles. PMID- 11414830 TI - Chiral recognition by CD-sensitive dimeric zinc porphyrin host. 1. Chiroptical protocol for absolute configurational assignments of monoalcohols and primary monoamines. AB - A general microscale protocol for the determination of absolute configurations of primary amino groups or secondary hydroxyl groups linked to a single stereogenic center is described. The chiral substrates are linked to the achiral trifunctional bidentate carrier molecule (3-aminopropylamino)acetic acid (1, H(2)NCH(2)CH(2)CH(2)NHCH(2)COOH) and the resultant conjugates are then complexed with dimeric zinc porphyrin host 2 giving rise to 1:1 host/guest sandwiched complexes. These complexes exhibit exciton-coupled bisignate CD spectra due to stereodifferentiation leading to preferred porphyrin helicity. Since the chiral sense of twist between the two porphyrins in the complex is dictated by the stereogenic center of the substrate, the sign of the couplet determines the absolute configuration at this center. The twist of the porphyrin tweezer in the complex can be predicted from the relative steric sizes of the groups flanking the stereogenic center, such that the bulkier group protrudes from the complex sandwich. In certain alpha-hydroxy esters and alpha-amino esters, electronic factors and hydrogen bonding govern the preferred conformation of the complex, and hence the CD spectra. PMID- 11414831 TI - Chiral recognition by CD-sensitive dimeric zinc porphyrin host. 2. Structural studies of host-guest complexes with chiral alcohol and monoamine conjugates. AB - A structural study of complexes formed between a dimeric zinc porphyrin tweezer (host) and chiral monoalcohols and monoamines derivatized by a bidentate carrier molecule (guest) confirmed that their CD couplets arise from the preferred porphyrin helicity of 1:1 host-guest complexes. NMR experiments and molecular modeling of selected tweezer complexes revealed that the preferred conformation is the one in which the L (larger) group protrudes from the porphyrin sandwich; this preferred helicity of the complex determines the CD of the complexes. It was found that the porphyrin ring-current induced (1)H chemical shifts and molecular modeling studies of the complex lead to the assignments of relative steric size of the L (large)/M (medium) substituents attached to the stereogenic center. The assignments, in turn, are correlated with the sign of the CD exciton couplet that establishes the absolute configuration at the stereogenic center. Variable temperature NMR experiments proved that the observed increase in CD amplitude at lower temperatures derives from conformational changes in the preferred offset geometry between two porphyrin rings. PMID- 11414832 TI - Stiff, and sticky in the right places: the dramatic influence of preorganizing guest binding sites on the hydrogen bond-directed assembly of rotaxanes. AB - Structural rigidity and the preorganization of thread binding sites are shown to have a major influence on template efficiency in the synthesis of hydrogen bond assembled rotaxanes. Preorganization is so effective, in fact, that with good hydrogen bond acceptors (amides) a "world record" yield of 97% for a [2]rotaxane is obtained. The truly remarkable feature of this efficient template, however, is that it allows even poor hydrogen bond acceptors (esters) to be used to prepare hydrogen bond-assembled rotaxanes, despite the presence of competing hydrogen bonding groups (anions) which bind the key intermediates at least 10000x more strongly than single, unorganized, ester groups! The structures of the rotaxanes are established unambiguously in solution by (1)H NMR spectroscopy and in the solid state by X-ray crystallography. As a series they provide unique experimental information regarding the nature of amide-ester hydrogen bonding interactions; in particular they suggest that in CDCl(3), amide-ester NH...O=C hydrogen bonds are approximately 1 kcal mol(-)(1) weaker than the corresponding amide-amide interactions. PMID- 11414833 TI - Heck reaction catalyzed by PD-modified zeolites. AB - [Pd]-exchanged NaY zeolites have been prepared, characterized, and applied for the first time for catalytic carbon-carbon coupling reactions. The catalysts exhibit a high activity and selectivity toward the Heck reaction of aryl bromides with olefins for small palladium concentrations (< or =0.1 mol % of Pd). The catalysts can easily be separated from the reaction mixture and reused after washing without loss in activity. No limitation to the diffusion of adducts in the zeolite cages was observed (for linear alkenes). The electronic nature of the aryl bromides and the olefins has a dominating effect on the reaction yield and selectivity. The heterogeneous catalysts quantitatively convert all types of all aryl bromide (complete conversion of bromobenzene within 30 min) and activated aryl chlorides under standard reaction conditions. Product form selectivity is observed in the Heck reaction with cyclic olefins. PMID- 11414834 TI - Equilibria between alpha- and beta-agostic stabilized rotamers of secondary alkyl niobium complexes. AB - The isopropyl chloro complex Tp(Me2)NbCl(i-Pr)(PhC&tbd1;CMe) (2) [Tp(Me2) = hydrotris(3,5-dimethylpyrazolyl)borate] exhibits a beta-agostic structure in the crystal. The conformation of the alkyl group is such that the agostic methyl group lies in the Calpha-Nb-Cl plane and the nonagostic one, in a wedge formed by two pyrazole rings. As observed by solution NMR spectroscopy, restricted rotation about the Nb-C bond allows the observation of an equilibrium between this species, 2beta, and a minor alpha-agostic rotamer 2alpha. A putative third rotamer which would have the secondary hydrogen in the wedge is not observed. Similar behavior is observed for related Tp'NbCl(i-Pr)(R(2)C=CMe) [Tp' = Tp(Me2), R(2) = Me (3); Tp' = Tp(Me2,4Cl), R(2) = Ph (4)]. The two diastereomers of the sec-butyl complex Tp(Me2)NbCl(sec-Bu)(MeC=CMe) (5) have been separated. In the crystal, 5CR-AS has a beta-agostic methyl group with the ethyl group located in the wedge formed by two pyrazole rings. The same single beta-agostic species is observed in solution. The other diastereomer, 5AR-CS has a beta-agostic methylene group in the solid state, and the methyl group sits in the wedge. In solution, an equilibrium between this beta-agostic methylene complex 5AR-CSbeta and a minor alpha-agostic species 5AR-CSalpha, where the ethyl substituent of the sec-Bu group is located in the wedge between two pyrazole rings, is observed. NMR techniques have provided thermodynamic parameters for these equilibria (K = 2beta/2alpha = 4.0 +/- 0.1 at 193 K, DeltaG(o)(193) = -2.2 +/- 0.1, DeltaH(o) = 7.4 +/- 0.1 kJ mol(-)(1), and DeltaS(o) = -27 +/- 1 J K(-)(1) mol(-)(1)), as well as kinetic parameters for the rotation about the Nb-C bond (at 193 K, DeltaG(2)= 47.5 +/- 2.5, DeltaH= 58.8 +/- 2.5 kJ mol(-)(1), and DeltaS = 59.0 +/- 10 J K( )(1) mol(-)(1)). Upon selective deuteration of the beta-methyl protons in Tp(Me2)NbCl[CH(CD(3))(2)](PhC=CMe) (2-d(6)), an expected isotope effect that displaces the equilibrium toward the alpha-agostic rotamer is observed (K = 2 d(6)beta/2-d(6)alpha = 3.1 +/- 0.1 at 193 K, DeltaG(o)(193) = -1.8 +/- 0.1, DeltaH(o) = -8.3 +/- 0.4 kJ mol(-)(1) and DeltaS(o)= -34 +/- 2 J K(-)(1) mol( )(1)). The anomalous values for DeltaH(o) and DeltaS(o) are discussed. Hybrid quantum mechanics/molecular mechanics calculations (IMOMM (B3LYP:MM3)) on the realistic model Tp(Me2)NbCl(i-Pr)(HC=CMe) have reproduced the energy differences between the alpha- and beta-agostic species with remarkable accuracy. Similar calculations show that Tp(Me2)NbCl(CH(2)Me)(HC=CMe) is alpha-agostic only and that Tp(5)(-)(Me)NbCl(CH(2)Me)(HC=CMe), which has no methyl groups at the 3 positions of the pyrazole rings, is beta-agostic only. Analysis and discussion of the computational and experimental data indicate that the unique behavior observed for the secondary alkyl complexes stems from competition between electronic effects favoring a beta-agostic structure and steric effects directing a bulky substituent in the wedge between two pyrazole rings of Tp(Me2). All of the secondary alkyl complexes thermally rearrange to the corresponding linear alkyl complexes via a first-order reaction. PMID- 11414835 TI - Issues of microscopic reversibility and an isomeric intermediate in ligand substitution reactions of five-coordinate oxorhenium(V) dithiolate complexes. AB - Ligand substitution reactions between five-coordinate oxorhenium(V) dithiolates, [CH(3)ReO(SCH(2)C(6)H(4)S)X], or MeReO(mtp)X, and entering ligands Y have been studied; Y is a phosphine and X is a phosphine (usually) or a pyridine. Many of them occur in two distinct stages, and other two-stage reactions merge to a single kinetic term when the successive rate constants are quite different in value. An intermediate can be detected directly by electronic and NMR spectroscopy. Just for phosphines, the range of rate constants is remarkably large; in the first stage, k spans the range 10(-)(4)-10(1) L mol(-)(1) s(-)(1) at 25 degrees C in benzene; in the second, which also shows a first-order dependence on the concentration of the entering ligand, the range is 10(-)(4) 10(3) L mol(-)(1) s(-)(1). Spectroscopic evidence shows that the intermediate has the same composition as the product; the metastable form is designated as MeReO(mtp)Y. The structures of all the isolated products MeReO(mtp)Y have a single stereochemistry: Me and -SCH(2) lie in trans positions, as do Y and -SAr. This structure is believed to be reversed in the transient, Y and -SCH(2) occupying trans positions. Further support for this assignment comes from the (31)P splitting of the (1)H NMR spectrum, where additional coupling indicates unusual four-bond coupling from a W-pattern of the hydrogen and phosphorus atoms. The intermediate does not undergo an intramolecular rearrangement to the final product; instead, it reacts with a ligand of the same type in an intermolecular reaction leading to rearrangement. The activation parameters were determined for selected reactions, and the results support a mechanism with considerable associative character; DeltaS() values are ca. -125 J K(-)(1) mol(-)(1). Because ligand Y must enter the coordination sphere from the vacant coordination position trans to the Re=O group, a means must be devised for the leaving group X to gain that position. To account for the intervention of the isomer while honoring the principle of microscopic reversibility, two mechanisms are proposed. One involves a C(3) ("turnstile") rotation of a specific group of three ligands in the six coordinate transition state. Turnstile rotation of the groups X, Me, and Y can accomplish the needed transposition; the transition state passes through an approximate trigonal prismatic configuration, giving rise to a different and less stable isomer. The alternative mechanism, which may more easily accommodate data for Y = Me(2)bpy, involves rearrangement of the common octahedral intermediate to a pentagonal pyramid. The arrangement of ligands in the intermediate, governed by their sizes, determines that isomerization accompanies product formation. Following either rearrangement, a second reaction, between MeReO(SCH(2)C(6)H(4)S)Y and Y, then ensues by the same mechanism. The second rearrangement process then generates the more stable isomer of the product. Results are also presented from a study of monomerization of the dimeric rhenium species, [MeReO(mtp)](2), with phosphines(X) of various size and basicity. The results support a mechanism with two intermediates on the pathway to MeReO(mtp)X. PMID- 11414836 TI - Phenylthiyl radical complexes of gallium(III), iron(III), and cobalt(III) and comparison with their phenoxyl analogues. AB - Three hexadentate, asymmetric pendent arm macrocycles containing a 1,4,7 triazacyclononane-1,4-diacetate backbone and a third, N-bound phenolate or thiophenolate arm have been synthesized. In [L(1)](3)(-) the third arm is 3,5-di tert-butyl-2-hydroxybenzyl, in [L(2)](3)(-) it is 2-mercaptobenzyl, and in [L(3)](3)(-) it is 3,5-di-tert-butyl-2-mercaptobenzyl. With trivalent metal ions these ligands form very stable neutral mononuclear complexes [M(III)L(1)] (M = Ga, Fe, Co), [M(III)L(2)] (M = Ga, Fe, Co), and [M(III)L(3)] (M = Ga, Co) where the gallium and cobalt complexes possess an S = 0 and the iron complexes an S = (5)/(2) ground state. Complexes [CoL(1)].CH(3)OH.1.5H(2)O, [CoL(3)].1.17H(2)O, [FeL(1)].H(2)O, and [FeL(2)] have been characterized by X-ray crystallography. Cyclic voltammetry shows that all three [M(III)L(1)] complexes undergo a reversible, ligand-based, one-electron oxidation generating the monocations [M(III)L(1)(*)](+) which contain a coordinated phenoxyl radical as was unambiguously established by their electronic absorption, EPR, and Mossbauer spectra. In contrast, [M(III)L(2)] complexes in CH(3)CN solution undergo an irreversible one-electron oxidation where the putative thiyl radical monocationic intermediates dimerize with S-S bond formation yielding dinuclear disulfide species [M(III)L(2)-L(2)M(III)](2+). [GaL(3)] behaves similarly despite the steric bulk of two tertiary butyl groups at the 3,5-positions of the thiophenolate, but [Co(III)L(3)] in CH(2)Cl(2) at -20 to -61 degrees C displays a reversible one-electron oxidation yielding a relatively stable monocation [Co(III)L(3)(*)](+). Its electronic spectrum displays intense transitions in the visible at 509 nm (epsilon = 2.6 x 10(3) M(-)(1) cm(-)(1)) and 670sh, 784 (1.03 x 10(3)) typical of a phenylthiyl radical. The EPR spectrum of this species at 90 K proves the thiyl radical to be coordinated to a diamagnetic cobalt(III) ion (g(iso) = 2.0226; A(iso)((59)Co) = 10.7 G). PMID- 11414837 TI - Artificial cytochrome b: computer modeling and evaluation of redox potentials. AB - We generated atomic coordinates of an artificial protein that was recently synthesized to model the central part of the native cytochrome b (Cb) subunit consisting of a four-helix bundle with two hemes. Since no X-ray structure is available, the structural elements of the artificial Cb were assembled from scratch using all known chemical and structural information available and avoiding strain as much as possible. Molecular dynamics (MD) simulations applied to this model protein exhibited root-mean-square deviations as small as those obtained from MD simulations starting with the crystal structure of the native Cb subunit. This demonstrates that the modeled structure of the artificial Cb is relatively rigid and strain-free. The model structure of the artificial Cb was used to determine the redox potentials of the two hemes by calculating the electrostatic energies from the solution of the linearized Poisson-Boltzmann equation (LPBE). The calculated redox potentials agree within 20 meV with the experimentally measured values. The dependence of the redox potentials of the hemes on the protein environment was analyzed. Accordingly, the total shift in the redox potentials is mainly due to the low dielectric medium of the protein, the protein backbone charges, and the salt bridges formed between the arginines and the propionic acid groups of the hemes. The difference in the shift of the redox potentials is due to the interactions with the hydrophilic side chains and the salt bridges formed with the propionic acids of the hemes. For comparison and to test the computational procedure, the redox potentials of the two hemes in the native Cb from the cytochrome bc(1) (Cbc(1)) complex were also calculated. Also in this case the computed redox potentials agree well with experiments. PMID- 11414838 TI - alpha- and 3(10)-helix interconversion: a quantum-chemical study on polyalanine systems in the gas phase and in aqueous solvent. AB - Helices are among the predominant secondary structures in globular proteins. About 90% of the residues in them are found to be in the alpha-helical conformation, and another 10% in the 3(10) conformation. There is a standing controversy between experimental and some theoretical results, and controversy among theoretical results concerning the predominance of each conformation, in particular, helices. We address this controversy by ab initio Hartree-Fock and density functional theory studies of helices with different lengths in a vacuum and in the aqueous phase. Our results show that (1) in a vacuum, all oligo(Ala) helices of 4-10 residues adopt the 3(10) - conformation; (2) in aqueous solution, the 6-10 residue peptides adopt the alpha-helical conformation; (3) there might be two intermediates between these helical conformers allowing for their interconversion. The relevance of these results to the structure and folding of proteins is discussed. PMID- 11414839 TI - Carbomethoxychlorocarbene: spectroscopy, theory, chemistry and kinetics. AB - Photolysis (254 nm) of methyl 8-chloro-3a,7a-methanoindan-8-carboxylate (5) in argon at 14 K produces carbomethoxychlorocarbene (6) as a persistent species. The IR and UV-vis spectra of the carbene were recorded and analyzed with the aid of density functional calculations (B3-LYP/6-31G). The IR spectrum of 6 is consistent with the carbene having a nonplanar singlet ground state, in agreement with the G3(MP2)//B3-LYP calculations of Scott and Radom (accompanying paper). Irradiation (300 nm) of 5 in solution produces indane in 97% yield. In cyclohexane, carbene 6 is trapped by insertion into a CH bond, whereas in 2,3 dimethylbutene it adds to the double bond to form a cyclopropane. Laser flash photolysis of 5 (308 nm, 17 ns, XeCl excimer) produces carbene 6 which reacts with pyridine to form an ylide (lambda(max) = 440 nm). It was possible to resolve the growth of the ylide in Freon-113 (CF(2)ClCFCl(2)) to measure the lifetime (tau = 114 ns, ambient temperature) of the carbene and the absolute rate constant of its reaction with pyridine (k(pyr) = 2 x 10(9) M(-)(1) s(-)(1)). A plot of log(1/tau) versus 1/T in CF(2)ClCFCl(2) is linear with Arrhenius parameters E(a) = 10.9 +/- 0.8 kJ/mol and A = 10(9.1)(+/-)(0.2) s(-)(1). In perfluorohexane, a less reactive solvent than Freon-113, E(a) = 23.4 +/- 1.7 kJ/mol, A = 10(10.6)(+/ )(0.) s(-)(1), and tau = 354 ns at 293 K. It is argued that the activation barrier to carbene disappearance in perfluorohexane represents the lower limit to the barrier to Wolff rearrangement of the carbene. PMID- 11414840 TI - Singlet-triplet splittings and barriers to Wolff rearrangement for carbonyl carbenes. AB - High-level ab initio calculations at the G3(MP2)//B3-LYP level have been used to study carbomethoxychlorocarbene and related halogenocarbenes and carbonyl carbenes. Initial calculations at the more accurate W1' level on the subset CH(2), HCCl, HCF, CCl(2), and CF(2) provide support for the reliability of G3(MP2)//B3-LYP for this type of problem. The W1' calculations also suggest that the experimental S-T splitting is slightly underestimated for HCCl and CF(2) and substantially underestimated for CCl(2), in keeping with other recent high-level studies. Whereas the parent carbonyl carbenes, namely formylcarbene, carbohydroxycarbene, and carbomethoxycarbene, are all predicted to have triplet ground states, their chloro and fluoro derivatives are predicted to have singlet ground states. In particular, carbomethoxychlorocarbene is predicted to have a singlet ground state, with the singlet-triplet splitting estimated as -16.0 kJ mol(-)(1). The barriers to Wolff rearrangement of the singlet carbonyl carbenes generally (but not always) correlate with the exothermicity accompanying the production of ketenes. In the case of the parent carbonyl carbenes, for which the rearrangement reaction is most exothermic, the barriers lie between about 10 and 30 kJ mol(-)(1), whereas for the less exothermic rearrangements of the chloro- and fluoro-substituted carbonyl carbenes, the Wolff rearrangement barriers increase significantly to between 58 and 75 kJ mol(-)(1). The calculated barrier for carbomethoxychlorocarbene is 58.2 kJ mol(-)(1). PMID- 11414841 TI - Troposelective substitutions in microsolvated systems. AB - The mechanism and the stereochemistry of the intracomplex "solvolysis" of the proton-bound complexes I(X)() between CH(3)(18)OH and (R)-(+)-1-aryl-ethanol (1(R)()(X)(); aryl = phenyl (X = H); pentafluorophenyl (X = F)) have been investigated in the gas phase in the 25-100 degrees C temperature range. The results point to intracomplex "solvolysis" as proceeding through the intermediacy of the relevant benzyl cation III(X)() (a pure S(N)1 mechanism). "Solvolysis" of I(H)() leads to complete racemization at T > 50 degrees C, whereas at T < 50 degrees C the reaction displays a preferential retention of configuration. Predominant retention of configuration is also observed in the intracomplex "solvolysis" of I(F)(). This picture is rationalized in terms of different intracomplex interactions between the benzylic ion III(X)() and the nucleophile/leaving group pair, which govern the timing of their reorientation within the electrostatic complex. The obtained gas-phase picture is discussed in the light of related gas-phase and solution data. It is concluded that the solvolytic reactions are mostly governed by the lifetime and the dynamics of the species involved and, if occurring in solution, by the nature of the solvent cage. Their rigid subdivision into the S(N)1 and S(N)2 mechanistic categories appears inadequate, and the use of their stereochemistry as a mechanistic probe can be highly misleading. PMID- 11414842 TI - Polyelectrolyte multilayers prepared from water-soluble poly(alkoxythiophene) derivatives. AB - Electronically conducting polyanion and polycation based on poly(alkoxythiophene) derivatives, poly-3-(3'-thienyloxy)propanesulfonate (P3TOPS) and poly-3-(3' thienyloxy)propyltriethylammonium (P3TOPA) have been synthesized. Both polymers are water-soluble and exhibit high conjugation length in solution and in the solid state. These polyelectrolytes were used to prepare conducting and electroactive polyelectrolyte multilayers by the sequential layer-by-layer adsorption technique. In aqueous solutions multilayers of P3TOPS with inactive polyelectrolytes (e.g., poly(diallyldimethylammonium chloride), PDADMA) displayed electrochemical and optical behavior similar to polythiophene films prepared in organic media. Their in-plane conductivity was low (ca. 1.6 x 10(-)(5) S cm( )(1)). The conductivity could, however, be increased by a factor of ca. 40 in "all-thiophene" films, in which P3TOPA was substituted for the inactive polycation (PDADMA). The interpenetration of layers is of prime importance in films containing conducting components. The interpenetration of P3TOPS was studied by measuring the charge-transfer rate across an insulating polyelectrolyte multilayer between the substrate and the P3TOPS layer with modulated electroreflectance. The extent of interpenetration was 8-9 polyelectrolyte layers, the length scale (7-15 nm) depending on the nature of the insulating layer and, especially, on the ionic strength of the solution used for the adsorption of P3TOPS. PMID- 11414843 TI - Solvent effects on methyl transfer reactions. 2. The reaction of amines with trimethylsulfonium salts. AB - The reaction of ammonia and pyridine with trimethylsulfonium ion has been studied in gas phase and solution. Density functional theory at the B3LYP/6-31+G level was used to describe the energy changes along the reaction coordinate in the gas phase, and the self-consistent isodensity polarizable continuum model (SCI-PCM) was used to calculate the effect of cyclohexane and dimethyl sulfoxide as the solvent on the energy changes. The effect of water as the solvent was studied using the Monte Carlo free energy perturbation method. The reaction with both ammonia and pyridine follows a similar rather convoluted path in gas phase, with the formation of several reaction complexes before and after the formation of the transition state. All the species found in gas phase persist in cyclohexane, yielding a reaction path very similar to that in gas phase but with significant differences in the relative energy of the critical points. In DMSO, the energy profile is greatly simplified by the disappearance of several of the species found in gas phase and in cyclohexane. The activation free energy increases with the polarity of the solvent in both reactions. Increasing the polarity of the solvent also increases the exothermicity of the reaction of trimethylsulfonium ion with ammonia and reduces it in the reaction with pyridine. In water, the free energy profile follows the same trend as found for DMSO, and free energy of activation is calculated to be larger by about 2-3 kcal/mol. This is in good agreement with an experimental measurement of the effect of solvent on the rate of reaction. PMID- 11414844 TI - Model-free approach to the dynamic interpretation of residual dipolar couplings in globular proteins. AB - The effects of internal motions on residual dipolar NMR couplings of proteins partially aligned in a liquid-crystalline environment are analyzed using a 10 ns molecular dynamics (MD) computer simulation of ubiquitin. For a set of alignment tensors with different orientations and rhombicities, MD-averaged dipolar couplings are determined and subsequently interpreted for different scenarios in terms of effective alignment tensors, average orientations of dipolar vectors, and intramolecular reorientational vector distributions. Analytical relationships are derived that reflect similarities and differences between motional scaling of dipolar couplings and scaling of dipolar relaxation data (NMR order parameters). Application of the self-consistent procedure presented here to dipolar coupling measurements of biomolecules aligned in different liquid-crystalline media should allow one to extract in a "model-free" way average orientations of dipolar vectors and specific aspects of their motions. PMID- 11414845 TI - Group epitope mapping by saturation transfer difference NMR to identify segments of a ligand in direct contact with a protein receptor. AB - A protocol based on saturation transfer difference (STD) NMR spectra was developed to characterize the binding interactions at an atom level, termed group epitope mapping (GEM). As an example we chose the well-studied system of galactose binding to the 120-kDa lectin Ricinus communis agglutinin I (RCA(120)). As ligands we used methyl beta-D-galactoside and a biantennary decasaccharide. Analysis of the saturation transfer effects of methyl beta-D-galactoside showed that the H2, H3, and H4 protons are saturated to the highest degree, giving evidence of their close proximity to protons of the RCA(120) lectin. The direct interaction of the lectin with this region of the galactose is in excellent agreement with results obtained from the analysis of the binding specificities of many chemically modified galactose derivatives (Bhattacharyya, L.; Brewer, C. F. Eur. J. Biochem. 1988, 176, 207-212). This new NMR technique can identify the binding epitope of even complex ligands very quickly, which is a great improvement over time-consuming chemical modifications. Efficient GEM benefits from a relatively high off rate of the ligand and a large excess of the ligand over the receptor. Even for a ligand like the biantennary decasaccharide with micromolar binding affinity, the binding epitopes could easily be mapped to the terminal beta-D-Gal-(1-4)-beta-D-GlcNAc (beta-D-GlcNAc = N-acetyl-D-glucosamine) residues located at the nonreducing end of the two carbohydrate chains. The binding contribution of the terminal galactose residue is stronger than those of the penultimate GlcNAc residues. We could show that the GlcNAc residues bind "edge-on" with the region from H2 to H4, making contact with the protein. Analysis of STD NMR experiments performed under competitive conditions proved that the two saccharides studied bind at the same receptor site, thereby ruling out unspecific binding. PMID- 11414846 TI - Solid-state (13)C NMR chemical shift anisotropy tensors of polypeptides. AB - Carbon-13 chemical shift anisotropy (CSA) tensors for various carbon sites of polypeptides, and for carbon sites in alpha-helical and beta-sheet conformations of poly-L-alanine, and polyglycine, are presented. The carbonyl (13)C CSA tensors were determined from one-dimensional CPMAS spectra obtained at a slow spinning speed, whereas the CSA tensors of C(alpha) and other carbons in side chains of peptides were determined using 2D PASS experiments on powder samples. The results suggest that the spans of (13)Carbonyl CSA tensors of alanine and glycine residues in various peptides are similar, even though the magnitude of individual components of the CSA tensor and the isotropic chemical shift are different. In addition, the delta(22) element is the only component of the (13)Carbonyl CSA tensor that significantly depends on the CO.HN hydrogen-bond length. Solid-state NMR experimental results also suggest that (13)Carbonyl and (13)C(alpha) CSA tensors are similar for alpha-helical and beta-sheet conformations of poly-L alanine, which is in agreement with the reported quantum chemical calculation studies and previous solid-state NMR experimental studies on other systems. On the other hand, the (13)C(alpha) CSA tensor of the first alanine residue is entirely different from that of the second or later alanine residues of the peptide. While no clear trends in terms of the span and the anisotropic parameter were predicted for (13)C(beta) CSA tensors of alanine, they mainly depend on the conformation and dynamics of the side chain as well as on the packing interactions in the solid state of peptides. PMID- 11414847 TI - The ozonolysis of acetylene--a quantum chemical investigation. AB - The ozonolysis of acetylene was investigated using CCSD(T), CASPT2, and B3LYP-DFT in connection with a 6-311+G(2d,2p) basis set. The reaction is initiated by the formation of a van der Waals complex followed by a [4pi + 2pi] cycloaddition between ozone and acetylene (activation enthalpy DeltaH(a)(298) = 9.6 kcal/mol; experiment, 10.2 kcal/mol), yielding 1,2,3-trioxolene, which rapidly opens to alpha-ketocarbonyl oxide 5. Alternatively, an O atom can be transferred from ozone to acetylene (DeltaH(a)(298) = 15.6 kcal/mol), thus leading to formyl carbene, which can rearrange to oxirene or ketene. The key compound in the ozonolysis of acetylene is 5 because it is the starting point for the isomerization to the corresponding dioxirane 19 (DeltaH(a)(298) = 16.9 kcal/mol), for the cyclization to trioxabicyclo[2.1.0]pentane 10 (DeltaH(a)(298) = 19.5 kcal/mol), for the formation of hydroperoxy ketene 15 (DeltaH(a)(298) = 20.6 kcal/mol), and for the rearrangement to dioxetanone 9 (DeltaH(a)(298) = 23.6 kcal/mol). Compounds 19, 10, 15, and 9 rearrange or decompose with barriers between 13 and 16 kcal/mol to yield as major products formanhydride, glyoxal, formaldehyde, formic acid, and (to a minor extent) glyoxylic acid. Hence, the ozonolysis of acetylene possesses a very complicated reaction mechanism that deserves intensive experimental studies. PMID- 11414848 TI - The anomalous infrared amide I intensity distribution in (13)C isotopically labeled peptide beta-sheets comes from extended, multiple-stranded structures: an ab initio study. AB - Ab initio based calculations of force fields and atomic polar tensors are used to simulate amide I infrared absorption spectra for a series of isotopically substituted (Ac-A(12)-NH-CH(3))(n)() peptides clustered in an antiparallel beta sheet conformation having a varying number of strands, n = 2-5. The results demonstrate that the anomalous intensity previously reported for the isotopically shifted amide I in (13)C labeled peptides is due to formation of multistranded beta-sheet structures in this conformation. Computations show that the characteristic widely split amide I mode for beta-sheet polypeptides as well as this anomalous intensity enhancement in isotopically substituted beta-sheet peptides grows with increasing sheet size. For sheets of five strands, qualitative and near quantitative agreement with experimental amide I intensity patterns is obtained for both labeled and unlabeled peptides. The strongest transitions primarily represent in-phase coupled modes of the (13)C labeled, next nearest neighbor amides on the inner strands of the multistranded beta-sheet. Long-range transition dipole coupling interactions do not promote the (13)C amide I intensity enhancement. Understanding of the IR intensity mechanisms with this level of detail for the isotopically labeled peptides permits design of site specific probes of beta-sheet folding and unfolding dynamics. PMID- 11414849 TI - Hypoiodous acid as guest molecule in protonated water clusters: a combined FT ICR/DFT study of I(H2O)n+. AB - Cationic water clusters containing iodine, of the composition I(H2O)n+, n = 0-25, are generated in a laser vaporization source and investigated by FT-ICR mass spectrometry. An investigation of blackbody radiation-induced fragmentation of size-selected clusters I(H2O)n+, n = 3-15, under collision-free conditions revealed an overall linear increase of the unimolecular rate constant with cluster size, similar to what has been observed previously for other hydrated ions. Above a certain critical size, I(H2O)n+, n greater than or approx. 13, reacts with HCl by formation of the interhalide ICl and a protonated water cluster, which is the reverse of a known solution-phase reaction. Accompanying density functional calculations illustrate the conceptual differences between cationic and anionic iodine-water clusters I(H2O)n+/-. While I-(H2O)n is genuinely a hydrated iodide ion, the cationic closed-shell species I(H2O)n+ may be best viewed as a protonated water cluster, in which one water molecule is replaced by hypoiodous acid. In the strongly acidic environment, HOI is protonated because of its high proton affinity. However, similar to the well known H3O+/H5O2+ controversy in protonated water clusters, a smooth transition between H2IO+ and H4IO2+ as core ions is observed for different cluster sizes. PMID- 11414850 TI - Mechanism of olefin cyclopropanation by diazomethane catalyzed by palladium dicarboxylates: a density functional study. AB - The reaction of diazomethane with ethylene in the presence of palladium diformate has been studied through density functional calculations. Several mechanistic paths leading to the formation of cyclopropane have been studied. The results obtained show that the reaction of palladium diformate with diazomethane is more favorable than the reaction with ethylene. The reaction with diazomethane may lead to two different isomeric complexes: a methylene-inserted complex and a palladium-carbene complex. Insertion of methylene is the most favorable process, but the resulting complex is not suitable for cyclopropanation. The reaction with two additional diazomethane molecules makes the formation of the bismethylene inserted palladium-carbene complex favorable. Attack of ethylene on this palladium-carbene complex leads to the formation of cyclopropane. PMID- 11414851 TI - Comparison of methyl rotation axis order parameters derived from model-free analyses of (2)H and (13)C longitudinal and transverse relaxation rates measured in the same protein sample. AB - Recombinant HIV-1 protease was obtained from bacteria grown on a 98% D(2)O medium containing 3-(13)C pyruvic acid as the sole source of (13)C and (1)H. The purified protein is highly deuterated at non-methyl carbons, but contains significant populations of (13)CHD(2) and (13)CH(2)D methyl isotopomers. This pattern of isotope labeling permitted measurements of (1)H and (13)C relaxation rates of (13)CHD(2) isotopomers and (2)H (D) relaxation rates of (13)CH(2)D isotopomers using a single sample. The order parameters S(axis)(2), which characterize the motions of the methyl rotation axes, were derived from model free analyses of R(1) and R(2) data sets measured for (13)C and (2)H spins. Our primary goal was to compare the S(axis)(2) values derived from the two independent types of data sets to test our understanding of the relaxation mechanisms involved. However, S(axis)(2) values derived from the analyses depend strongly on the geometry of the methyl group, the sizes of the quadrupolar and dipolar couplings, and the effects of bond vibrations and librations on these couplings. Therefore uncertainties in these basic physical parameters complicate comparison of the order parameters. This problem was circumvented by using an experimental relationship, between the methyl quadrupolar, (13)C-(13)C and (13)C (1)H dipolar couplings, derived from independent measurements of residual static couplings of weakly aligned proteins by Ottiger and Bax (J. Am. Chem. Soc. 1999, 121, 4690-4695) and Mittermaier and Kay (J. Am. Chem. Soc. 1999, 121, 10608 10613). This approach placed a tight experimental restraint on the values of the (2)H quadrupolar and (13)C-(1)H dipolar interactions and greatly facilitated the accurate comparison of the relative values of the order parameters. When applied to our data this approach yielded satisfactory agreement between the S(axis)(2) values derived from the (13)C and (2)H data sets. PMID- 11414852 TI - Identification of defect sites on MgO(100) thin films by decoration with Pd atoms and studying CO adsorption properties. AB - CO adsorption on Pd atoms deposited on MgO(100) thin films has been studied by means of thermal desorption (TDS) and Fourier transform infrared (FTIR) spectroscopies. CO desorbs from the adsorbed Pd atoms at a temperature of about 250 K, which corresponds to a binding energy, E(b), of about 0.7 +/- 0.1 eV. FTIR spectra suggest that at saturation two different sites for CO adsorption exist on a single Pd atom. The vibrational frequency of the most stable, singly adsorbed CO molecule is 2055 cm(-)(1). Density functional cluster model calculations have been used to model possible defect sites at the MgO surface where the Pd atoms are likely to be adsorbed. CO/Pd complexes located at regular or low-coordinated O anions of the surface exhibit considerably stronger binding energies, E(b) = 2 2.5 eV, and larger vibrational shifts than were observed in the experiment. CO/Pd complexes located at oxygen vacancies (F or F(+) centers) are characterized by much smaller binding energies, E(b) = 0.5 +/- 0.2 or 0.7 +/- 0.2 eV, which are in agreement with the experimental value. CO/Pd complexes located at the paramagnetic F(+) centers show vibrational frequencies in closest agreement with the experimental data. These comparisons therefore suggest that the Pd atoms are mainly adsorbed at oxygen vacancies. PMID- 11414853 TI - First oxadiazole-functionalized terbium(III) beta-diketonate for organic electroluminescence. PMID- 11414854 TI - Generation of homogeneous (sp(3)-C(1))-bridged Cp/amido and Cp/phosphido group 4 metal Ziegler-Natta catalyst systems. PMID- 11414855 TI - Dioxygen reactivity of mononuclear heme and copper components yielding a high spin heme-peroxo-cu complex. PMID- 11414856 TI - Chiral scaffolds for enantiocontrolled synthesis: enantio- and regiocontrolled [4+2] cycloaddition to 3-alkenyl-n(3)-pyranylmolybdenum complexes. PMID- 11414857 TI - Probing the effect of the outer saccharide residues of N-linked glycans on peptide conformation. PMID- 11414858 TI - Significant chemo- and regioselectivies in the Ru(II)-catalyzed [2+2+2] cycloaddition of 1,6-diynes with dicyanides. PMID- 11414859 TI - Paterno-Buchi reactions of allylic alcohols and acetates with aldehydes: hydrogen bond interaction in the excited singlet and triplet states? PMID- 11414860 TI - High-pressure synthesis of amorphous MgNi(1.02)H(2.2). PMID- 11414861 TI - A highly enantioselective chiral Lewis base-catalyzed asymmetric cyanation of ketones. PMID- 11414862 TI - Dramatic enhancement of activities for living Ziegler-Natta polymerizations mediated by "exposed" zirconium acetamidinate initiators: the isospecific living polymerization of vinylcyclohexane. PMID- 11414863 TI - Lewis base activation of Lewis acids: catalytic enantioselective allylation and propargylation of aldehydes. PMID- 11414864 TI - Cyclodextrin-mediated soft cutting of single-walled carbon nanotubes. PMID- 11414865 TI - C-H bond activation of external substrates with a bis(mu-oxo)dicopper(III) complex. PMID- 11414866 TI - A simple bifunctional fluoroionophore signaling different metal ions either independently or cooperatively. PMID- 11414867 TI - NH tautomerism of N-confused porphyrin. PMID- 11414868 TI - Electron-accepting system of Si-Si bond in linear framework by combination with strong donor. PMID- 11414869 TI - Chiral 15-metallacrown-5 complexes differentially bind carboxylate anions. PMID- 11414870 TI - Controlled multi-stage recognition of guests using orthogonal electro- and photochemical inputs. PMID- 11414871 TI - Role of religion in the lives of Latino families of young children with developmental delays. AB - We interviewed 250 parents of Mexican and Puerto Rican origin living in the United States who had young children with developmental delays to determine the role of religion in their lives. Quantitative results indicate that parents largely viewed themselves as religious, were affiliated with a formal religion, and participated in religious activities. Most parents viewed both church and faith as supportive, but faith was shown to provide more support. Repeated measures a analysis of variance found some intragroup variations in religious support and changes in support after learning of the child's condition. Thematic analysis revealed specific religious beliefs and practices parents viewed as supportive, and content and cultural models analyses indicated the religious frameworks by which parents interpreted their child's disability. PMID- 11414872 TI - Enhancing free-recall rates of individuals with mental retardation. AB - Generative encoding contexts promote activation of multiple retrieval routes and have been shown to enhance free-recall rates of individuals without mental retardation. The present extension to individuals with mental retardation involved a comparison of two encoding conditions: (a) fade-in, initially presenting pictures out of focus then slowly fading them into focus, and (b) fade out, presenting pictures clearly then slowly blurring them. Results indicated that free-recall rates were greater for the fade-in items for the individuals with mental retardation and CA-matched comparisons, but not for the MA-matched group. These findings demonstrate the utility of a generative encoding context that does not involve verbal instruction for individuals with and without mental retardation. PMID- 11414873 TI - Intentional communication elicits language-facilitating maternal responses in dyads with children who have developmental disabilities. AB - An experimental test of whether intentional communication elicits maternal responses purported to facilitate language development is presented. Fifty-eight prelinguistic children with disabilities and their mothers participated. Children were randomly assigned to two staff-implemented treatments that were designed to experimentally increase intentional communication ability. We predicted that treatment effects on maternal responses would vary by maternal education level and maternal responsive interaction style at the pretreatment period. We predicted any treatment effect on maternal responses would be mediated by posttreatment intentional communication. Results indicated that treatment effects on maternal responses varied by pretreatment maternal education level. A pattern of results also supports the conclusion that posttreatment intentional communication to mother was, at least in part, responsible for the effect on posttreatment maternal responses. PMID- 11414874 TI - Preliminary study of altered skin temperature at body sites associated with self injurious behavior in adults who have developmental disabilities. AB - In this study, the sensory status of 4 nonverbal adults with mental retardation and severe self-injury was examined using skin temperature measures prior to opiate antagonist treatment. Double-blind, placebo-controlled, experimental ABAB designs were used to evaluate the effects of naltrexone hydrochloride (1.5 mg/kg/day). For each participant, the body site targeted most frequently for self injury was associated with altered skin temperature and reduced by naltrexone. In all cases, neither infrequent self-injury body sites nor non-self-injury body sites were associated with altered skin temperature. Further controlled studies are warranted to examine the value of assessing pain status and skin temperature in nonverbal patients with mental retardation and related developmental disabilities who present with tissue-damaging SIB. PMID- 11414875 TI - Evaluation of the dissemination of active support training in staffed community residences. AB - Our aim was to replicate Active Support, a staff training intervention designed to increase the assistance given to adults with severe mental retardation living in community residences in order to increase their participation in activities. Training was conducted in 38 residences, involving 303 staff members and 106 persons with mental retardation. Active Support resulted in significant increases in assistance and engagement in activity. Significant correlations between participant abilities, receipt of assistance, and levels of engagement were found. Active Support was found to be of greater benefit to people with lower adaptive behavior and to have a diminishing value for people with higher adaptive behavior, implying that its implementation should be matched to the support needs of residents. PMID- 11414876 TI - Intellectual classification of Black and White children in special education programs using the WISC-III and the Cognitive Assessment System. AB - The Wechsler Intelligence Scale for Children--Third Edition and the Cognitive Assessment System were compared for a sample of 78 White and Black students in special education programs for children with mental retardation. Results showed that the WISC-III identified more children as having mental retardation than did the Cognitive Assessment System. More important, however, the WISC-III classified disproportionately more Blacks than Whites as having mental retardation as compared to the Cognitive Assessment System. Results imply that the problem of disproportionate representation of Black children in special education classes for children with mental retardation may be addressed if the Cognitive Assessment System were used instead of the WISC-III. PMID- 11414877 TI - Measuring physical inclusion of people with developmental disabilities: evaluation of the Macomb-Oakland Regional Center. AB - The Macomb-Oakland Regional Center has been facilitating the movement of persons with developmental disabilities into the community for 27 years. To measure randomness of their home locations, we used a geographic-based spatial analytic approach. Involvement of allies in planning futures, number of people with disabilities residing in each home, proportion of earned income, level of disability, and level of mobility were used as explanatory variables. Monte Carlo procedures using Moran's I, a measure of spatial autocorrelation, did not reveal significant clustering of residences. Regression procedures, however, did reveal that earned income was the independent variable that most explained the degree to which people were physically included. People living in smaller homes were more randomly dispersed. PMID- 11414878 TI - Density functional theory for colloidal rod-sphere mixtures. AB - We present a density functional theory for a model colloidal mixture of hard spheres and infinitely thin hard rods. For these freely rotating particles, we use a fundamental measures approach to obtain a functional that features the correct dimensional crossover and the exact low density limit. For isotropic bulk fluid mixtures, the free energy, and hence the demixing phase diagram, are identical to that obtained from free volume theory. Results for the partial pair correlation functions of the bulk mixture are in good agreement with those of our simulations. PMID- 11414879 TI - Dynamics of electrostatically driven granular media: effects of humidity. AB - We performed experimental studies of the effect of humidity on the dynamics of electrostatically driven granular materials. Both conducting and dielectric particles undergo a phase transition from an immobile state (granular solid) to a fluidized state (granular gas) with increasing applied field. Spontaneous precipitation of solid clusters from the gas phase occurs as the external driving is decreased. The clustering dynamics in conducting particles is primarily controlled by screening of the electric field but is aided by cohesion due to humidity. It is shown that humidity effects dominate the clustering process with dielectric particles. PMID- 11414880 TI - Field-driven helix unwinding in antiferroelectric liquid crystal cells. AB - The interaction between the antiferroelectric helix and an electric field applied perpendicular to its axis is investigated, both experimentally and theoretically. A two-stage switching process is observed: (i) in the pretransitional regime the helix distorts and then unwinds to form a nonhelical antiferroelectric state, with the plane of the molecules parallel to the applied field; (ii) at higher fields switching to the ferroelectric state occurs. The mechanism for unwinding is the interaction of the applied field with a polarization that is induced by a change to the anticlinic ordering. PMID- 11414881 TI - Casimir problem of spherical dielectrics: quantum statistical and field theoretical approaches. AB - The Casimir free energy for a system of two dielectric concentric nonmagnetic spherical bodies is calculated with use of a quantum statistical mechanical method, at arbitrary temperature. By means of this rather novel method, which turns out to be quite powerful (we have shown this to be true in other situations also), we consider first an explicit evaluation of the free energy for the static case, corresponding to zero Matsubara frequency (n=0). Thereafter, the time dependent case is examined. For comparison we consider the calculation of the free energy with use of the more commonly known field theoretical method, assuming for simplicity metallic boundary surfaces. PMID- 11414882 TI - Behavior of the reaction front between initially segregated species in a two stage reaction. AB - The large-time asymptotic behavior of a two-stage reaction (A+B-->R, B+R-->S) with initially segregated reactants is described. The concentration of the reactants is found to be significantly less than the initial concentrations in a depletion zone of width proportional to t(1/2), where t is time; the reaction takes place in a thinner zone of width proportional to t(1/6). Similarity solutions for the chemical concentration profiles in the reaction zone are calculated, and are compared with numerical simulations of the full partial differential reaction-diffusion equations. The large-time asymptotic scalings reported here are the same as in the absence of the secondary reaction, but we find that the location of the reaction zone is significantly shifted due to the secondary reaction. The reaction zone may behave in an exotic fashion at large time, moving first one way, then reversing its direction. PMID- 11414883 TI - Reunion of random walkers with a long range interaction: applications to polymers and quantum mechanics. AB - We use a renormalization group to calculate the reunion and survival exponents of a set of random walkers interacting with a long range 1/r2 and a short range interaction. These exponents are used to study the binding-unbinding transition of polymers and the behavior of several quantum problems. PMID- 11414884 TI - Monte Carlo simulations for a Lotka-type model with reactant surface diffusion and interactions. AB - The standard Lotka-type model, which was introduced for the first time by Mai et al. [J. Phys. A 30, 4171 (1997)] for a simplified description of autocatalytic surface reactions, is generalized here for a case of mobile and energetically interacting reactants. The mathematical formalism is proposed for determining the dependence of transition rates on the interaction energy (and temperature) for the general mathematical model, and the Lotka-type model, in particular. By means of Monte Carlo computer simulations, we have studied the impact of diffusion (with and without energetic interactions between reactants) on oscillatory properties of the A+B-->2B reaction. The diffusion leads to a desynchronization of oscillations and a subsequent decrease of oscillation amplitude. The energetic interaction between reactants has a dual effect depending on the type of mobile reactants. In the limiting case of mobile reactants B the repulsion results in a decrease of amplitudes. However, these amplitudes increase if reactants A are mobile and repulse each other. A simplified interpretation of the obtained results is given. PMID- 11414885 TI - Fluctuation theorem for Hamiltonian systems: Le Chatelier's principle. AB - For thermostated dissipative systems, the fluctuation theorem gives an analytical expression for the ratio of probabilities that the time-averaged entropy production in a finite system observed for a finite time takes on a specified value compared to the negative of that value. In the past, it has been generally thought that the presence of some thermostating mechanism was an essential component of any system that satisfies a fluctuation theorem. In the present paper, we point out that a fluctuation theorem can be derived for purely Hamiltonian systems, with or without applied dissipative fields. PMID- 11414886 TI - Diffusion in stationary flow from mesoscopic nonequilibrium thermodynamics. AB - We analyze the diffusion of a Brownian particle in a fluid under stationary flow. By using the scheme of nonequilibrium thermodynamics in phase space, we obtain the Fokker-Planck equation that is compared with others derived from the kinetic theory and projector operator techniques. This equation exhibits violation of the fluctuation-dissipation theorem. By implementing the hydrodynamic regime described by the first moments of the nonequilibrium distribution, we find relaxation equations for the diffusion current and pressure tensor, allowing us to arrive at a complete description of the system in the inertial and diffusion regimes. The simplicity and generality of the method we propose makes it applicable to more complex situations, often encountered in problems of soft condensed matter, in which not only one but more degrees of freedom are coupled to a nonequilibrium bath. PMID- 11414887 TI - Monostable array-enhanced stochastic resonance. AB - We present a simple nonlinear system that exhibits multiple distinct stochastic resonances. By adjusting the noise and coupling of an array of underdamped, monostable oscillators, we modify the array's natural frequencies so that the spectral response of a typical oscillator in an array of N oscillators exhibits N 1 different stochastic resonances. Such families of resonances may elucidate and facilitate a variety of noise-mediated cooperative phenomena, such as noise enhanced propagation, in a broad class of similar nonlinear systems. PMID- 11414888 TI - Percolation and jamming in random sequential adsorption of linear segments on a square lattice. AB - We present the results of a study of random sequential adsorption of linear segments (needles) on sites of a square lattice. We show that the percolation threshold is a nonmonotonic function of the length of the adsorbed needle, showing a minimum for a certain length of the needles, while the jamming threshold decreases to a constant with a power law. The ratio of the two thresholds is also nonmonotonic and it remains constant only in a restricted range of the needles length. We determine the values of the correlation length exponent for percolation, jamming, and their ratio. PMID- 11414889 TI - Escape time in anomalous diffusive media. AB - We investigate the escape behavior of systems governed by the one-dimensional nonlinear diffusion equation theta(t)rho=theta(x)[theta(x)Urho]+Dtheta(x)2rho(nu), where the potential of the drift, U(x), presents a double well and D,nu are real parameters. For systems close to the steady state, we obtain an analytical expression of the mean first passage time, yielding a generalization of Arrhenius law. Analytical predictions are in very good agreement with numerical experiments performed through integration of the associated Ito-Langevin equation. For nu not equal to 1, important anomalies are detected in comparison to the standard Brownian case. These results are compared to those obtained numerically for initial conditions far from the steady state. PMID- 11414890 TI - Noise-assisted transmission of binary information: theory and experiment. AB - We study the response of a bistable vertical cavity surface emitting laser to an aperiodic binary signal, by adding a variable amount of noise. The resulting behavior is an example of aperiodic stochastic resonance, and in this work we give a detailed comparison between analytical and numerical results and accurate experimental measurements. We characterize the phenomenon by using different appropriate indicators, which also allow us to quantify the binary information transmission. We show that the quality of the transmission is enhanced by a suitable amount of noise, and we give a physical picture of the phenomenon. PMID- 11414891 TI - Strong enhancement of noise-induced escape by nonadiabatic periodic driving due to transient chaos. AB - We have found a mechanism by which a moderately weak nonadiabatic periodic driving may significantly facilitate noise-induced interwell transitions in an underdamped multiwell system. The mechanism is associated with the onset of a homoclinic tangle in the noise-free system: if the ratio of the driving amplitude A to the damping gamma exceeds a critical value approximately 1, then the basins of attraction of the linear responses related to different wells are mixed in a complex manner in some layer associated with the separatrix of the undriven nondissipative system, and the minimal energy in such layer is lower than the top of the barrier. Thus the energy to which the system needs to be activated by the noise, to be able to make a transition, is lower than the top of the barrier. PMID- 11414892 TI - Preferential growth: exact solution of the time-dependent distributions. AB - We consider a preferential growth model where particles are added one by one to the system consisting of clusters of particles. A new particle can either form a new cluster (with probability q) or join an already existing cluster with a probability proportional to the size thereof. We calculate exactly the probability Pi(k,t) that the size of the ith cluster at time t is k. We analyze the asymptotics, the scaling properties of the size distribution and of the mean size, as well as the relation of our system to recent network models. PMID- 11414893 TI - Generalized fundamental solutions for unsteady viscous flows. AB - A number of closed-form fundamental solutions for generalized unsteady Oseen and Stokes flows associated with arbitrary time-dependent translational and rotational motions have been developed. These solutions are decomposed into two parts corresponding to a longitudinal wave and a transversal wave. As examples of application, the hydrodynamic forces acting on a sphere and on a circular cylinder translating in an unsteady rotating flow field at low Reynolds numbers are calculated using the generalized fundamental solutions. PMID- 11414894 TI - Collective dynamics in liquid lead: generalized propagating excitations. AB - A microscopic approach to the investigation of generalized collective excitations, developed recently for pure liquids, is applied to the study of the spectrum of collective excitations in a liquid metal. The calculations are performed for liquid lead at two temperatures (above the melting point and in high-temperature region) and the results are compared. From the analysis of spectra, obtained for different basis sets of dynamical variables, we conclude that there exist three branches of propagating collective excitations, which correspond to sound and heat (high- and low-frequency) waves in the liquid. It is shown that the branch of low-frequency heat waves contains a propagation gap in the hydrodynamic region. An analytical expression for the width of the propagation gap for low-frequency heat waves is derived. PMID- 11414895 TI - Transformation from Rogers-Young approximation to the density functional approach for nonuniform fluids: numerical recipe. AB - A numerical recipe is devised to extend the methodology [J. Chem. Phys. 112, 8079 (2000)] to nonhard-sphere nonuniform fluids where analytical expressions for the functional relationship of the bridge function as a function of indirect correlation function do not exist, the numerical recipe is also based on the universality of the free-energy density functional. As an example, the recipe is employed to calculate the density profile of a colloidal suspension near a single charged hard wall and the hard-sphere Yukawa fluid near a single hard wall and a single hard wall with an attractive tail, the agreement of the predictions of the theory with the simulation data is good. The difference of the present methodology from that of the weighted density approximation is discussed. PMID- 11414896 TI - Edwards' measures: a thermodynamic construction for dense granular media and glasses. AB - We present numerical support for the hypothesis that macroscopic observables of dense granular media and glasses can be evaluated from averages over typical blocked configurations: we construct the corresponding measure for a class of finite-dimensional systems and compare its predictions for various observables with the outcome of the out of equilibrium dynamics at large times. We discuss in detail the connection with the effective temperatures that appear in out of equilibrium glass theories, as well as the relation between our computation and those based on "inherent structure" arguments. A short version of this work has appeared in Phys. Rev. Lett. 85, 5034 (2000). PMID- 11414897 TI - Glassy systems under time-dependent driving forces: application to slow granular rheology. AB - We study the dynamics of a glassy model with infinite range interactions externally driven by an oscillatory force. We find a well-defined transition in the (temperature-amplitude-frequency) phase diagram between (i) a "glassy" state characterized by the slow relaxation of one-time quantities, aging in two-time quantities and a modification of the equilibrium fluctuation-dissipation relation; and (ii) a "liquid" state with a finite relaxation time. In the glassy phase, the degrees of freedom governing the slow relaxation are thermalized to an effective temperature. Using Monte Carlo simulations, we investigate the effect of trapping regions in phase space on the driven dynamics. We find that it alternates between periods of rapid motion and periods of trapping. These results confirm the strong analogies between the slow granular rheology and the dynamics of glasses. They also provide a theoretical underpinning to earlier attempts to present a thermodynamic description of moderately driven granular materials. PMID- 11414898 TI - Large-scale synchrony in weakly interacting automata. AB - We study the behavior of two spatially distributed (sandpile) models which are weakly linked with one another. Using a Monte Carlo implementation of the renormalization-group and algebraic methods, we describe how large-scale correlations emerge between the two systems, leading to synchronized behavior. PMID- 11414899 TI - Numerical model for granular compaction under vertical tapping. AB - A simple numerical model is used to simulate the effect of vertical taps on a packing of monodisperse hard spheres. Our results are in good agreement with an experimental work done in Chicago and with other previous models, especially concerning the dynamics of the compaction, the influence of the excitation strength on the compaction efficiency, and some aging effects. The principal asset of the model is that it allows a local analysis of the packings. Vertical and transverse density profiles are used, as well as size and volume distributions of the pores. An interesting result concerns the appearance of a vertical gradient in the density profiles during compaction. Furthermore, the volume distribution of the pores suggests that the smallest pores, ranging in size between tetrahedral and octahedral sites, are not strongly affected by the tapping process, in contrast to the largest pores which are more sensitive to the compaction of the packing. PMID- 11414900 TI - Colloids confined to a flexible container. AB - A model of hard spheres trapped inside a container of fluctuating shape is proposed to describe colloidal particles in a vesicle or in an emulsion droplet. The container is assumed to be the convex hull of the particles and is described by an integral geometric approach including volume and surface terms. In the limit of large volume coupling, the model reduces to the well-known geometric problem of natural bin packing. Using computer simulations and cell theory, we calculate equilibrium properties for various finite numbers of confined particles in conformations ranging from clusters to planar and linear structures and identify transitions between these different conformations. PMID- 11414901 TI - Dynamic structure of thermoreversible colloidal gels of adhesive spheres. AB - The dynamic structure factor f(q,t) of suspensions of adhesive colloidal spheres has been characterized as a function of temperature over the volume fraction range 0.0100, reaching at least 40% of the measured intensity. Both the phase diagram and the low-k structural data of dense Kr turn out to be closely reproduced by the hierarchical reference theory if additional many-body forces are taken into account by an augmented strength of the Axilrod-Teller triple-dipole potential. The experimental density derivative of S(k) is also used for a very sensitive test of the theories and interaction models considered here. PMID- 11414943 TI - Nonlinear ac response of an electrorheological fluid. AB - The applied electric field used in most electrorheological (ER) experiments is usually quite high, and nonlinear ER effects have been measured recently. When a nonlinear ER fluid is subjected to a sinusoidal (ac) field, the electrical response will in general consist of ac fields at frequencies of the higher-order harmonics. In this paper, a self-consistent formalism has been employed to compute the induced dipole moment for ER fluids in which the suspended particles have nonlinear characteristics, in an attempt to investigate the ac response of a nonlinear ER fluid. PMID- 11414944 TI - Quasicritical behavior of the low-frequency dielectric permittivity in the isotropic phase of liquid crystalline materials. AB - Results presented give evidence of the existence of quasicritical, fluidlike behavior in the isotropic phase of 4-cyano-4-pentyl-biphenyl (5CB) for frequencies ranging from the static to the ionic-dominated [low-frequency (LF)] region. Despite the boost of dielectric permittivity on lowering the frequency below 1 kHz, values of the isotropic-nematic transition discontinuity (approximately 1.1 K) and the critical exponent alpha (approximately 0.5) remain constant. It is shown that the contribution from residual ionic impurities is a linear function of temperature in the critical, prenematic fluctuation-dominated region. The validity of the fluidlike and critical behavior for LF dielectric permittivity confirmed results of a derivative analysis of the experimental data: d(epsilon)/dT proportional to (T-T*)(-alpha), originally proposed for critical mixtures. Results of a preliminary test in the isotropic phase of 4-decyl-4' isothiocyanatobiphenyl (10BT), on approaching the smectic-E phase, may indicate a general validity of results obtained. PMID- 11414945 TI - Influence of topological constraints on the statics and dynamics of ring polymers. AB - We report a computer simulation study of the influence of topological constraints on the statics and dynamics of single ring polymers and ring polymers in the melt. We show that single rings have identical static and dynamic scaling behavior regardless of the presence of topological constraints. For rings in the melt we find that the scaling behavior is significantly influenced by the presence of topological constraints. PMID- 11414946 TI - Small numerators canceling small denominators: is Dyson's hierarchical model solvable? AB - We present an analytical method to solve Dyson's hierarchical model, involving the scaling variables near the high-temperature fixed point. The procedure seems plagued by small denominators as in perturbative expansions near integrable systems in Hamiltonian mechanics. However, in 36 cases considered, a zero denominator always comes with a zero numerator. We conjecture that these cancellations occur in general, allowing the application of the analytical method and suggesting that the model has remarkable features reminiscent of the integrable systems. PMID- 11414947 TI - Coupled length scales in eroding landscapes. AB - We report results from an empirical study of the anisotropic structure of eroding landscapes. By constructing a novel correlation function, we show quantitatively that small-scale channel-like features of landscapes are coupled to the large scale structure of drainage basins. We show additionally that this two-scale interaction is scale-dependent. The latter observation suggests that a commonly applied effective equation for erosive transport may itself depend on scale. PMID- 11414948 TI - Boundary lubrication: dynamics of squeeze-out. AB - The dynamics of the expulsion of the last liquid monolayer of molecules confined between two surfaces (measured recently for the first time) has been analyzed by solving the two-dimensional Navier-Stokes equation combined with kinetic Monte Carlo simulations. Instabilities in the boundary line of the expelled film were observed. We show that the instabilities produce a rough boundary for all length scales above a critical value and a smooth boundary for shorter lengths. The squeezing out of all but a few trapped islands of liquid is shown to be the result of the pressure gradient in the contact area. PMID- 11414949 TI - Crossover behavior in the isothermal susceptibility near the 3He critical point. AB - We present high-resolution measurements of the isothermal susceptibility of pure 3He near the liquid-gas critical point. PVT measurements were performed in the single-phase region over the reduced temperature range 3 x 10(-5)2omega+omega) in which the multistep process 2omega=omega+omega breaks the phase invariance of the down-conversion process. A parametrically-forced Ginzburg-Landau equation is derived to explain the existence of phase-armed spiral waves. PMID- 11414952 TI - Loss of synchronization in coupled oscillators with ubiquitous local stability. AB - The issue of using instantaneous eigenvalues as indicators of synchronization quality in coupled chaotic systems is examined. Previously, it has been assumed that, if the eigenvalues of the linearized synchronization dynamics have negative real parts everywhere on the attractor, the synchronized state is stable. In this Rapid Communication, two counterexamples are presented that show this assumption is invalid. PMID- 11414953 TI - Interface imprinting by a rippled shock using an intense laser. AB - Perturbation imprinting at a flat interface by a rippled shock has been observed in a laser hydrodynamics experiment. A strong shock was driven through a three layer target, with the first interface rippled, and the second flat. The chosen thickness of the second layer gave instability growth with opposite phases at the two interfaces, consistent with two-dimensional simulations and rippled shock theory. PMID- 11414954 TI - Stable localized vortex solitons. AB - We demonstrate that parametric interaction of a fundamental beam with its second harmonic in bulk media, in the presence of self-defocusing third-order nonlinearity, gives rise to the first ever examples of completely stable localized ring-shaped solitons with intrinsic vorticity n=1 and n=2. The stability is demonstrated both in direct simulations and by computing eigenvalues of the corresponding linearized equations. A potential application of the (2+1) dimensional ring solitons in optics is a possibility to design a reconfigurable multichannel system guiding signal beams. PMID- 11414955 TI - Statistical mechanics for unstable states in Gel'fand triplets and investigations of parabolic potential barriers. AB - Free energies and other thermodynamical quantities are investigated in canonical and grand canonical ensembles of statistical mechanics involving unstable states which are described by the generalized eigenstates with complex energy eigenvalues in the conjugate space of Gel'fand triplet. The theory is applied to the systems containing parabolic potential barriers (PPB's). The entropy and energy productions from PPB systems are studied. An equilibrium for a chemical process described by reactions A+CB <==>AC+B is also discussed. PMID- 11414956 TI - Surface critical behavior of random systems: ordinary transition. AB - We calculate the surface critical exponents of the ordinary transition occurring in semi-infinite, quenched dilute Ising-like systems. This is done by applying the field theoretic approach directly in d=3 dimensions up to the two-loop approximation as well as in 4-epsilon dimensions. At d=4-epsilon we extend, up to the next-to-leading order, the previous first-order results of the square root of epsilon expansion by Ohno and Okabe [Phys. Rev. B 46, 5917 (1992)]. In both cases numerical estimates for surface exponents are computed using Pade approximants extrapolating the perturbation theory expansions. The obtained results indicate that the critical behavior of semi-infinite systems with quenched bulk disorder is characterized by the new set of surface critical exponents. PMID- 11414957 TI - Fluctuation effects in an epidemic model. AB - We study a discrete epidemic model A+B-->2A in one and two dimensions (1D and 2D). In 1D for low concentration theta, we find that a depletion zone exists ahead of the front and the average velocity of the front approaches v=theta/2. In the 1D high concentration limit, we find that the velocity approaches v=1-e( theta/2). In 2D, for low concentration we also find a depletion zone, and the velocity scales as v approximately theta(0.6), which is different from the scaling expected from the mean field approximation, v approximately theta(0.5). Analysis of the interface width scaling properties demonstrated that the front dynamics of this reaction are not governed by the Kardar-Parisi-Zhang equation. PMID- 11414958 TI - Tracer dispersion in two-dimensional rough fractures. AB - Tracer diffusion and hydrodynamic dispersion in two-dimensional fractures with self-affine roughness are studied by analytic and numerical methods. Numerical simulations were performed via the lattice-Boltzmann approach, using a boundary condition for tracer particles that improves the accuracy of the method. The reduction in the diffusive transport, due to the fractal geometry of the fracture surfaces, is analyzed for different fracture apertures. In the limit of small aperture fluctuations we derive the correction to the diffusive coefficient in terms of the tortuosity, which accounts for the irregular geometry of the fractures. Dispersion is studied when the two fracture surfaces are simply displaced normally to the mean fracture plane and when there is a lateral shift as well. Numerical results are analyzed using the Lambda parameter, related to convective transport within the fracture, and simple arguments based on lubrication approximation. At very low Peclet number, in the case where fracture surfaces are laterally shifted, we show using several different methods that convective transport reduces dispersion. PMID- 11414959 TI - Dynamic instability of dislocations due to nucleation of a new phase. AB - Dislocation lines in a crystal close to a bulk phase transition can be coated by nuclei of a new phase which for a moving dislocation gives rise to a viscous friction force. In some range of the material parameters this force leads to a dynamic instability of the defect line, resulting from an intricate interplay between the shape fluctuations of the defect and the amplitude fluctuations of the nucleus. The instability shows up in the linear response of the dislocation to a periodic change of the driving force and in the structure factor of the order-parameter fluctuations of the nucleus. PMID- 11414960 TI - Standing localized cluster in a continuum traffic model. AB - We study the emergence of standing localized cluster in a continuum traffic model. The local-density profile, local velocity profile, and the phase boundary are obtained. The effect of the on ramp is discussed. The indication of the boundary induced phase transition is also discussed. PMID- 11414961 TI - Fractures in heterogeneous two-dimensional systems. AB - A two-dimensional triangular lattice with bond disorder is used as a testing ground for fracture behavior in heterogeneous materials in strain-controlled conditions. Simulations are performed with two interaction potentials (harmonic and Lennard-Jones types) and different breaking thresholds. We study the strain range where the fracture progressively develops from the first to the last breakdown. Scaling properties with the lattice size are investigated: no qualitative difference is found between the two interaction potentials. Clustering properties of the broken bonds are also studied by grouping them into disjoint sets of connected bonds. Finally, the role of kinetic energy is analyzed by comparing overdamped with dissipationless dynamics. PMID- 11414962 TI - Universal behavior of one-dimensional multispecies branching and annihilating random walks with exclusion. AB - A directed percolation process with two symmetric particle species exhibiting exclusion in one dimension is investigated numerically. It is shown that if the species are coupled by branching (A-->AB, B-->BA), a continuous phase transition will appear at the zero-branching-rate limit belonging to the same universality class as that of the two component branching and annihilating random-walk model with two symmetric offsprings. This class persists even if the branching is biased towards one of the species. If the two systems are not coupled by branching but a hard-core interaction is allowed only the transition will occur at finite branching rate belonging to the usual (1+1)-dimensional directed percolation class. PMID- 11414963 TI - Macroscopically frustrated Ising model. AB - A disordered spin-glass model in which both static and dynamical properties depend on macroscopic magnetizations is presented. These magnetizations interact via random couplings and, therefore, the typical quenched realization of the system exhibits a macroscopic frustration. The model is solved by using a revisited replica approach, and the broken symmetry solution turns out to coincide with the symmetric solution. Some dynamical aspects of the model are also discussed, showing how it could be a useful tool for describing some properties of real systems such as, for example, natural ecosystems or human social systems. PMID- 11414964 TI - Minimal current phase and universal boundary layers in driven diffusive systems. AB - We investigate boundary-driven phase transitions in open driven diffusive systems. The generic phase diagram for systems with short-ranged interactions is governed by a simple extremal principle for the macroscopic current, which results from an interplay of density fluctuations with the motion of shocks. In systems with more than one extremum in the current-density relation, one finds a minimal current phase even though the boundaries support a higher current. The boundary layers of the critical minimal current and maximal current phases are argued to be of a universal form. The predictions of the theory are confirmed by Monte Carlo simulations of the two-parameter family of stochastic particle hopping models of Katz, Lebowitz, and Spohn and by analytical results for a related cellular automaton with deterministic bulk dynamics. The effect of disorder in the particle jump rates on the boundary layer profile is also discussed. PMID- 11414965 TI - Thermodynamics and fractional Fokker-Planck equations. AB - The relaxation to equilibrium in many systems that show strange kinetics is described by fractional Fokker-Planck equations (FFPEs). These can be considered as phenomenological equations of linear nonequilibrium theory. We show that the FFPEs describe a system whose noise in equilibrium fulfills the Nyquist theorem. Moreover, we show that for subdiffusive dynamics, the solutions of the corresponding FFPEs are probability densities for all cases in which the solutions of the normal Fokker-Planck equation (with the same Fokker-Planck operator and with the same initial and boundary conditions) exist. The solutions of the FFPEs for superdiffusive dynamics are not always probability densities. This fact means only that the corresponding kinetic coefficients are incompatible with each other and with the initial conditions. PMID- 11414966 TI - Exact solution of a class of one-dimensional nonequilibrium stochastic models. AB - We consider various one-dimensional nonequilibrium models, namely, the diffusion limited pair-annihilation and creation model (DPAC) and its unbiased version (the Lushnikov model), the DPAC model with particle injection, as well as (biased) diffusion-limited coagulation model (DC). We study the DPAC model using an approach based on a duality transformation and the generating function of the dual model. We are able to compute exactly the density and correlation functions in the general case with arbitrary initial states. Further, we assume that a source injects particles in the system. Solving, via the duality transformation, the equations of motion of the density, and the noninstantaneous two-point correlation functions, we see how the source affects the dynamics. Finally we extend the previous results to the DC model with help of a similarity transformation. PMID- 11414967 TI - Three-loop critical exponents, amplitude functions, and amplitude ratios from variational perturbation theory. AB - We use variational perturbation theory to calculate various universal amplitude ratios above and below Tc in minimally subtracted straight phi4 theory with N components in three dimensions. In order to best exhibit the method as a powerful alternative to Borel resummation techniques, we consider only two- and three loops expressions where our results are analytic expressions. For the critical exponents, we also extend existing analytic expressions for two loops to three loops. PMID- 11414968 TI - Dynamic scaling in diluted systems: deactivation through thermal dilution. AB - Activated scaling is confirmed to hold in transverse field-induced phase transitions of randomly diluted Ising systems. Quantum Monte Carlo calculations have been made not just at the percolation threshold (pc) but also well below and above it. We follow the evolution of the activated scaling at zero temperature in the phase transition from ferromagnetic to quantum Griffiths phase (p>pc) at the phase boundary (p=pc) and for transitions inside the nonferromagnetic quantum Griffiths phase (pinfinity we calculate the location alphac of the phase transition (signaling the onset of anomalous response), and solve the statics for alpha>alphac exactly. The temporal correlations in global market fluctuations turn out not to decay to zero for infinitely widely separated times. For alpha0 we analyze our equations in leading order in alpha, and find asymptotic solutions with diverging volatility sigma=O(alpha(-1/2)) (as regularly observed in simulations), but also asymptotic solutions with vanishing volatility sigma=O(alpha(1/2)). The former, however, are shown to emerge only if the agents' initial strategy valuations are below a specific critical value. PMID- 11414976 TI - Highly optimized tolerance in epidemic models incorporating local optimization and regrowth. AB - In the context of a coupled map model of population dynamics, which includes the rapid spread of fatal epidemics, we investigate the consequences of two new features in highly optimized tolerance (HOT), a mechanism which describes how complexity arises in systems which are optimized for robust performance in the presence of a harsh external environment. Specifically, we (1) contrast global and local optimization criteria and (2) investigate the effects of time dependent regrowth. We find that both local and global optimization lead to HOT states, which may differ in their specific layouts, but share many qualitative features. Time dependent regrowth leads to HOT states which deviate from the optimal configurations in the corresponding static models in order to protect the system from slow (or impossible) regrowth which follows the largest losses and extinctions. While the associated map can exhibit complex, chaotic solutions, HOT states are confined to relatively simple dynamical regimes. PMID- 11414977 TI - Heterogeneous condensation in dense media. AB - A theoretical description of heterogeneous nucleation kinetics is presented. This description takes into account the perturbation of the vapor phase initiated by the growing droplets. The form of the density profile around the growing droplet is analyzed and some special approximations are given. Then the process of nucleation in the whole system is described. As a result all the main characteristics of the process are determined analytically. PMID- 11414978 TI - Turing pattern formation induced by spatially correlated noise. AB - The effect of spatially correlated noise on Turing structures is analyzed both experimentally and numerically. Using the photosensitive character of the chlorine dioxide-iodine-malonic acid reaction-diffusion system, spatial randomness is introduced in the system. In the presence of noise, Turing patterns appear and are stable at levels of average illumination that would be more than sufficient to suppress pattern formation in the case of homogeneous illumination. PMID- 11414979 TI - Scaling properties of scale-free evolving networks: continuous approach. AB - The scaling behavior of scale-free evolving networks, arising in areas such as communications, scientific citations, collaborations, etc., is studied. We derive universal scaling relations describing properties of such networks, and indicate the limits of their validity. We show that the main properties of scale-free evolving networks may be described in the framework of a simple continuous approach. The simplest models of networks, growing according to a mechanism of preferential attachment of links to nodes, are used. We consider different forms of this preference, and demonstrate that the range of preferential attachments producing scale-free networks is wide. We also obtain scaling relations for networks with nonlinear, accelerating growth, and describe the temporal evolution of the arising distributions. Size effects-the cutoffs of these distributions introduce restrictions for the observation of power-law dependences. Mainly we discuss the so-called degree distribution, i.e., the distribution of the number of connections of nodes. A scaling form of the distribution of links between pairs of individual nodes for a growing network of citations is also studied. We describe the effects of differences between nodes. The "aging" of nodes changes the exponents of the distributions. The appearance of a single node with high fitness changes the degree distribution of a network dramatically. If its fitness exceeds some threshold value, this node captures a finite part of all links of the network. We show that permanent random damage to a growing scale-free network a permanent deletion of some links-radically changes the values of the scaling exponents. Results of other kinds of permanent damage are described. PMID- 11414980 TI - Generation of unpredictable time series by a neural network. AB - A perceptron that "learns" the opposite of its own output is used to generate a time series. We analyze properties of the weight vector and the generated sequence, such as the cycle length and the probability distribution of generated sequences. A remarkable suppression of the autocorrelation function is explained, and connections to the Bernasconi model are discussed. If a continuous transfer function is used, the system displays chaotic and intermittent behavior, with the product of the learning rate and amplification as a control parameter. PMID- 11414981 TI - Minimal vertex covers on finite-connectivity random graphs: a hard-sphere lattice gas picture. AB - The minimal vertex-cover (or maximal independent-set) problem is studied on random graphs of finite connectivity. Analytical results are obtained by a mapping to a lattice gas of hard spheres of (chemical) radius 1, and they are found to be in excellent agreement with numerical simulations. We give a detailed description of the replica-symmetric phase, including the size and entropy of the minimal vertex covers, and the structure of the unfrozen component which is found to percolate at a connectivity c approximately 1.43. The replica-symmetric solution breaks down at c=e approximately 2.72. We give a simple one-step replica symmetry-broken solution, and discuss the problems in the interpretation and generalization of this solution. PMID- 11414982 TI - Effects of surfaces on resistor percolation. AB - We study the effects of surfaces on resistor percolation at the instance of a semi-infinite geometry. Particularly we are interested in the average resistance between two connected ports located on the surface. Based on general grounds as symmetries and relevance we introduce a field theoretic Hamiltonian for semi infinite random resistor networks. We show that the surface contributes to the average resistance only in terms of corrections to scaling. These corrections are governed by surface resistance exponents. We carry out renormalization-group improved perturbation calculations for the special and the ordinary transition. We calculate the surface resistance exponents phiS and phiS(infinity) for the special and the ordinary transition, respectively, to one-loop order. PMID- 11414983 TI - Theory of structure formation in snowfields motivated by penitentes, suncups, and dirt cones. AB - Penitentes and suncups are structures formed as snow melts, typically high in the mountains. When the snow is dirty, dirt cones and other structures can form instead. Building on previous field observations and experiments, this paper presents a theory of ablation morphologies, and the role of surface dirt in determining the structures formed. The glaciological literature indicates that sunlight, heating from air, and dirt all play a role in the formation of structure on an ablating snow surface. The present paper formulates a minimal model for the formation of ablation morphologies as a function of measurable parameters and considers the linear stability of this model. The dependence of ablation morphologies on weather conditions and initial dirt thickness is studied, focusing on the initial growth of perturbations away from a flat surface. We derive a single-parameter expression for the melting rate as a function of dirt thickness, which agrees well with a set of measurements by Driedger. An interesting result is the prediction of a dirt-induced traveling instability for a range of parameters. PMID- 11414984 TI - Linear and nonlinear information flow in spatially extended systems. AB - Infinitesimal and finite amplitude error propagation in spatially extended systems are numerically and theoretically investigated. The information transport in these systems can be characterized in terms of the propagation velocity of perturbations Vp. A linear stability analysis is sufficient to capture all the relevant aspects associated to propagation of infinitesimal disturbances. In particular, this analysis gives the propagation velocity VL of infinitesimal errors. If linear mechanisms prevail on the nonlinear ones Vp=VL. On the contrary, if nonlinear effects are predominant finite amplitude disturbances can eventually propagate faster than infinitesimal ones (i.e., Vp>VL). The finite size Lyapunov exponent can be successfully employed to discriminate the linear or nonlinear origin of information flow. A generalization of the finite size Lyapunov exponent to a comoving reference frame allows us to state a marginal stability criterion able to provide Vp both in the linear and in the nonlinear case. Strong analogies are found between information spreading and propagation of fronts connecting steady states in reaction-diffusion systems. The analysis of the common characteristics of these two phenomena leads to a better understanding of the role played by linear and nonlinear mechanisms for the flow of information in spatially extended systems. PMID- 11414985 TI - Integrability and action operators in quantum Hamiltonian systems. AB - For a (classically) integrable quantum-mechanical system with two degrees of freedom, the functional dependence H=HQ(J1,J2) of the Hamiltonian operator on the action operators is analyzed and compared with the corresponding functional relationship H(p1,q1;p2,q2)=HC(J1,J2) in the classical limit of that system. The former converges toward the latter in some asymptotic regime associated with the classical limit, but the convergence is, in general, nonuniform. The existence of the function H=HQ(J1,J2) in the integrable regime of a parametric quantum system explains empirical results for the dimensionality of manifolds in parameter space on which at least two levels are degenerate. The analysis is carried out for an integrable one-parameter two-spin model. Additional results presented for the (integrable) circular billiard model illuminate the same conclusions from a different angle. PMID- 11414986 TI - Quantum-classical correspondences of the Berry-Robnik parameter through bifurcations in lemon billiard systems. AB - The quantum level statistics affected by bifurcations in classical dynamics is studied by using a one-parameter family of lemon billiard systems. The classical phase space of our system consists of regular and irregular regions. We determine an analytic solution of the phase volume for these regions as a function of the system parameter and show that the function reveals a cusp singularity at the bifurcation point. The function is compared with its quantum mechanical counterpart, the Berry-Robnik parameter. By estimating the semiclassical regime from the effective Planck constant that validates the quantum-classical correspondence of the Berry-Robnik parameter, we determine a region of the system parameter where the cusp can be reproduced by the statistical properties of the eigenenergy levels. PMID- 11414987 TI - Is semiquantum chaos real? AB - A semiquantum system, composed of a quantum part coupled to a classical part, can exhibit dynamical chaos in the motion of the quantum state vector. However, there has been disagreement as to whether this mathematical chaos is physically real or merely an artifact of the semiquantum approximation. It is shown, for a model of a quantum spin coupled to an approximately classical nonlinear oscillator, that the semiquantum chaos disappears rapidly as the mass of the oscillator is increased to make it more classical. The time interval during which the semiquantum approximation remains accurate increases with the mass, but is not closely related to the Lyapunov time. PMID- 11414988 TI - Spatiotemporal nonlinear dynamics of a magnetoelastic ribbon. AB - Magnetoelastic materials have a strong coupling between strain and magnetization, so applying a magnetic field to a magnetoelastic material can change its shape. This coupling leads to interesting dynamics. We have studied the dynamics of a wide ribbon of Metglass 2605sc which was driven by a magnetic field. The ribbon was suspended as a pendulum in a set of Helmholtz coils, which provided both dc and ac magnetic fields. Laser light was reflected off the ribbon to measure its angular displacement. Two points on the ribbon could be simultaneously illuminated, and one of the laser beams could be scanned over the ribbon. We observed quasiperiodic bifurcations in the motion of the ribbon, and characterized the spatial aspect of the motion with some recently developed statistics. PMID- 11414989 TI - Regular dynamics of low-frequency fluctuations in external cavity semiconductor lasers. AB - It is commonly believed that the dynamics responsible for low-frequency fluctuations (LFF's) in external cavity semiconductor lasers is stochastic or chaotic. A common approach to address the origin of LFF's is to investigate the dynamical behavior of, and the interaction among, various external cavity modes in the Lang-Kobayashi (LK) paradigm. In this paper, we propose a framework for understanding of the LFFs based on a different set of fundamental solutions of the LK equations, which are periodic or quasiperiodic, and which are characterized by a sequence of time-locked pulses with slowly varying magnitude. We present numerical evidence and heuristic arguments, indicating that the dynamics of LFF's emerges as a result of quasiperiodic bifurcations from these solutions as the pumping current increases. Regular periodic solutions can actually be observed when (1) the feedback level is moderate, (2) pumping current is below solitary threshold, and (3) the linewidth enhancement factor is relatively large. PMID- 11414990 TI - Topological aspects of chaotic scattering in higher dimensions. AB - We investigate the topological properties of the chaotic invariant set associated with the dynamics of scattering systems with three or more degrees of freedom. We show that the separation of one degree of freedom from the rest in the asymptotic regime, a common property in a large class of scattering models, defines a gate which is a dynamical object with phase space separating invariant manifolds. The manifolds form an invariant set causing singularities in the scattering process. The codimension one property of the manifolds ensures that the fractal structure of the invariant set can be studied by scattering functions defined over simple one-dimensional families of initial conditions as usually done in two-degree-of freedom scattering problems. It is found that the fractal dimension of the invariant set is not due to the gates but to interior hyperbolic periodic orbits. PMID- 11414991 TI - Ergodicity and scars of the quantum cat map in the semiclassical regime. AB - We study the quantum localization effect of the cat map manifested in the motions of statistical ensembles. Specifically, the coarse-grained entropy and time averaged phase space distributions are investigated. For this purpose, an amended version of the Wigner function on the discretized phase torus is presented. We find that the time average of the coarse-grained Wigner function is scarred (antiscarred) along some short periodic orbits, and the heights (depths) of these scars (antiscars) decrease in a linear way with the Planck constant when the semiclassical limit is approached. The relationship between the scars observed here and those exhibited in the quasienergy eigenstates is discussed. PMID- 11414992 TI - Controlling neuronal spikes. AB - We propose two control strategies for achieving desired firing patterns in a physiologically realistic model neuron. The techniques are powerful, efficient, and robust, and we have applied them successfully to obtain a range of targeted spiking behaviors. The methods complement each other: one involves the manipulation of only a parameter, the applied soma current, and the other involves the manipulation of only a state variable, the membrane potential. Both techniques have the advantage that they are not measurement-intensive nor do they involve much run-time computation, as knowledge of only the interspike interval is necessary to implement control. PMID- 11414993 TI - Characterization and stabilization of the unstable fixed points of a frequency doubled Nd:YAG laser. AB - We demonstrate the successful stabilization of type II chaos of a multimode, intracavity frequency doubled, diode-pumped Nd:YAG (neodymium-doped yttrium aluminum garnet) laser in experiment using an adaptive proportional feedback control. The two orthogonal polarized infrared intensities are fed back to the injection current of the pump diode. The stability properties of the stabilized unstable fixed points are investigated and exploited to explain the performance of our control scheme and to determine suitable measurement signals for the feedback control. PMID- 11414994 TI - Taming chaos by impurities in two-dimensional oscillator arrays. AB - The effect of impurities in a two-dimensional lattice of coupled nonlinear chaotic oscillators and their ability to control the dynamical behavior of the system are studied. We show that a single impurity can produce synchronized spatiotemporal patterns, even though all oscillators and the impurity are chaotic when uncoupled. When a small number of impurities is arranged in a way, that the lattice is divided into two disjoint parts, synchronization is enforced even for small coupling. The synchronization is not affected as the size of the lattice increases, although the impurity concentration tends to zero. PMID- 11414995 TI - Multistability formation and synchronization loss in coupled Henon maps: two sides of the single bifurcational mechanism. AB - We investigate phenomena of multistability and complete chaos synchronization in coupled Henon maps, which is an invertible system. Multiparametric analysis of a selected family of periodic orbits for coupled Henon maps shows that a single bifurcational mechanism describes both a loss of chaos synchronization and multistability formation. The process of bubbling transition and riddle basins, and the multistability formation in invertible systems are described in detail. PMID- 11414996 TI - Pattern dynamics in bidimensional oscillatory media with bistable inhomogeneities. AB - By means of numerical simulations, we study pattern dynamics in selected examples of inhomogeneous active media described by a reaction diffusion model of the activator-inhibitor type. We consider inhomogeneities corresponding to a variation in space of the (nonlinear) reaction characteristics of the system or the diffusion constants. Three different bidimensional systems are analyzed: an oscillatory medium in a square reactor with a circular central bistable domain, and cases of a bistable stripe immersed in an oscillatory medium in a trapezoidal reactor and in a rectangular reactor with inhomogeneous diffusion. The different types of complex behavior that arise in these systems are analyzed. PMID- 11414997 TI - Temporal structures in shell models. AB - The intermittent dynamics of the turbulent Gledzer, Ohkitani, and Yamada shell model is completely characterized by a single type of burstlike structure, which moves through the shells like a front. This temporal structure is described by the dynamics of the instantaneous configuration of the shell amplitudes, revealing an approximate chaotic attractor of the dynamics. PMID- 11414998 TI - Reconstruction of the parameter spaces of dynamical systems. AB - Parameter variations in the equations of motion of dynamical systems are identified by time series analysis. The information contained in time series data is transformed and compressed to feature vectors. The space of feature vectors is an embedding for the unobserved parameters of the system. We show that the smooth variation of d system parameters can lead to paths of feature vectors on smooth d dimensional manifolds in feature space, provided the latter is high-dimensional enough. The number of varying parameters and the nature of their variation can thus be identified. The method is illustrated using numerically generated data and experimental data from electromotors. Complications arising from bifurcations in deterministic dynamical systems are shown to disappear for slightly noisy systems. PMID- 11414999 TI - Chaotic motion of space charge wave fronts in semiconductors under time independent voltage bias. AB - A standard drift-diffusion model of space charge wave propagation in semiconductors has been studied numerically and analytically under dc voltage bias. For sufficiently long samples, appropriate contact resistivity, and applied voltage-such that the sample is biased in a regime of negative differential resistance-we find chaos in the propagation of nonlinear fronts (charge monopoles of alternating sign) of electric field. The chaos is always low dimensional, but has a complex spatial structure; this behavior can be interpreted using a finite dimensional asymptotic model in which the front (charge monopole) positions and the electrical current are the only dynamical variables. PMID- 11415000 TI - Influence of inlet and bulk noise on Rayleigh-Benard convection with lateral flow. AB - Spatiotemporal properties of convective fluctuations and of their correlations are investigated theoretically in the vicinity of the threshold for onset of convection in the presence of a lateral through-flow using the full linearized equations of fluctuating hydrodynamics. The effect of external forcing by inlet boundary conditions on the downstream evolution of convective fields is separated from the effect of internal bulk thermal forcing with the use of spatial Laplace transformations. They show how the spatial variation of fluctuations and of their correlations are governed by the six spatial characteristic exponents of the field equations. PMID- 11415001 TI - Spectrum of anisotropic exponents in hydrodynamic systems with pressure. AB - We discuss the scaling exponents characterizing the power-law behavior of the anisotropic components of correlation functions in turbulent systems with pressure. The anisotropic components are conveniently labeled by the angular momentum index l of the irreducible representation of the SO(3) symmetry group. Such exponents govern the rate of decay of anisotropy with decreasing scales. It is a fundamental question whether they ever increase as l increases, or they are bounded from above. The equations of motion in systems with pressure contain nonlocal integrals over all space. One could argue that the requirement of convergence of these integrals bounds the exponents from above. It is shown here on the basis of a solvable model (the "linear pressure model") that this is not necessarily the case. The model introduced here is of a passive vector advection by a rapidly varying velocity field. The advected vector field is divergent free and the equation contains a pressure term that maintains this condition. The zero modes of the second-order correlation function are found in all the sectors of the symmetry group. We show that the spectrum of scaling exponents can increase with l without bounds while preserving finite integrals. The conclusion is that contributions from higher and higher anisotropic sectors can disappear faster and faster upon decreasing the scales also in systems with pressure. PMID- 11415002 TI - Growth rate of the linear Richtmyer-Meshkov instability when a shock is reflected. AB - An analytic model is presented to calculate the growth rate of the linear Richtmyer-Meshkov instability in the shock-reflected case. The model allows us to calculate the asymptotic contact surface perturbation velocity for any value of the incident shock intensity, arbitrary fluids compressibilities, and for any density ratio at the interface. The growth rate comes out as the solution of a system of two coupled functional equations and is expressed formally as an infinite series. The distinguishing feature of the procedure shown here is the high speed of convergence of the intermediate calculations. There is excellent agreement with previous linear simulations and experiments done in shock tubes. PMID- 11415003 TI - Effect of viscosity in the dynamics of two point vortices: exact results. AB - An exact, unstationary, two-dimensional solution of the Navier-Stokes equations for the flow generated by two point vortices is obtained. The viscosity nu is introduced as a Brownian motion in the Hamiltonian dynamics of point vortices. The point vortices execute a stochastic motion whose probability density can be computed from a Fokker-Planck equation, equivalent to the original Navier-Stokes equation. The derived solution describes, in particular, the merging process of two Lamb vortices, and the development of the characteristic spiral structure in the topology of the vorticity. The viscous effects are thoroughly investigated by an asymptotic analysis of the solution. In particular, the selection mechanism of a specific pattern among the infinity satisfying the nu=0 (Euler) equation is discussed. PMID- 11415004 TI - Reynolds stress under a change of frame of reference. AB - In this paper, we study the characteristics of the Reynolds stress under a change of frame, as defined by the Euclidean group of transformation. We show that being subject to the dynamical processes induced from the mean Navier-Stokes equations, the invariance group of the fluctuating velocity and the Reynolds stress is no longer the Euclidean group of transformation, which is merely a kinematical aspect, but reduces to the extended Galilean group of transformation. As a consequence, in contrast to developing the constitutive equations for the Cauchy stress in continuum mechanics, wherein the principle of material frame indifference is a guiding principle, the frame-dependent kinematical quantities, e.g., the mean spin tensor, may be allowed to play an effective role as the constitutive variable in turbulence modeling. PMID- 11415005 TI - Finite time singularities in a class of hydrodynamic models. AB - Models of inviscid incompressible fluid are considered, with the kinetic energy (i.e., the Lagrangian functional) taking the form L approximately integral k(alpha)/vk/2dk in 3D Fourier representation, where alpha is a constant, 0 approximately /x1-x2/(-alpha3), is calculated self-consistently in the nonuniversal region from which we obtain alpha3=3. Also we derive an equation governing the evolution of the probability density function (PDF) of longitudinal velocity increments in length scale, from which a possible mechanism for the dependence of the inertial PDF to one-point u(rms) is developed. PMID- 11415008 TI - Geometrical analysis of chaotic mixing in a low Reynolds number magnetohydrodynamic quadripolar flow. AB - A mixing device for highly viscous fluids with finite electrical conductivity is investigated theoretically. Stirring is performed by means of electromagnetic forces provided by inductor wires located outside the flow domain. The flow shows hyperbolic and elliptic singular points. Inductors are displaced in a periodic manner, leading to an efficient stretching and folding mechanism. The goodness of mixing is quantified by means of a geometrical analysis based on box-counting techniques. This analysis gives valuable information about advection of a spot of dye injected in the flow, in the limit of infinite Peclet numbers. A spatiotemporal criterion for mixing efficiency is derived, and characteristic scales are analyzed. The influence of various parameters on mixing efficiency is discussed by making use of both the geometrical analysis and Poincare sections. PMID- 11415009 TI - Heat transfer and convection onset in a compressible fluid: 3He near the critical point. AB - Heat transport in 3He above its critical temperature Tc was studied along the critical isochore in a flat Rayleigh-Benard cell (height h=1 mm, diameter D=57 mm). The range of the reduced temperature epsilon was 5 x 10(-4)< or = epsilon < or =2 x 10(-1). The temperature difference deltaT(t) across the fluid layer as a function of the time t was measured for different values of the heat current q until steady state was reached. The crossover was observed from the regime dominated by the Rayleigh criterion for the convection onset to that controlled by the adiabatic temperature gradient (ATG), or "Schwarzschild criterion," in good quantitative agreement with predictions. The slope of the convective heat current versus the reduced Rayleigh number was found to be independent of compressibility and the same as for still less compressible fluids. Plots of Nu versus Ra, both corrected for the ATG effect, are presented for early-stage convective turbulence (1 x 10(5) phi are obtained in one dimension via Monte Carlo simulation. The long-time behavior of these correlation functions clearly deviates from that of a recent analytical prediction of Bares and Mobilia [Phys. Rev. Lett. 83, 5214 (1999)]. An alternative expression for the asymptotic behavior is conjectured from numerical data. PMID- 11415047 TI - Stretched exponential relaxation in the mode-coupling theory for the Kardar Parisi-Zhang equation. AB - We study the mode-coupling theory for the Kardar-Parisi-Zhang equation in the strong-coupling regime, focusing on the long time properties. By a saddle point analysis of the mode-coupling equations, we derive exact results for the correlation function in the long-time limit-a limit that is hard to study using simulations. The correlation function at wave vector k in dimension d is found to behave asymptotically at time t as C(k,t) approximately A/k(d+4 2z)(Btk(z))(gamma/z)exp[-(Btk(z))(1/z)], with gamma=(d-1)/2, A a determined constant, and B a scale factor. PMID- 11415048 TI - Exact solution for the lattice gas model in one dimension. AB - A simple method to obtain a canonical partition function for a one-dimensional lattice gas model is presented. The simplification is based upon rewriting a sum over all possible configurations as a sum over all possible numbers of clusters in the system. PMID- 11415049 TI - Analysis of chaotic motion and its shape dependence in a generalized piecewise linear map. AB - We analyze the chaotic motion and its shape dependence in a piecewise linear map using Fujisaka's characteristic function method. The map is a generalization of the one introduced by Artuso. Exact expressions for diffusion coefficients are obtained with previously obtained results used as special cases. A fluctuation spectrum relating to the probability density function is obtained in a parametric form. We also give limiting forms of the above quantities. The dependence of the diffusion coefficient and probability density function on the shape of the map is examined. PMID- 11415050 TI - Topological defects after a quench in a Benard-Marangoni convection system. AB - We report experimental evidence of the fact that, in a Benard-Marangoni conduction-convection transition, the density of defects in the emerging structure scales as a power law in the quench time needed for the control parameter to ramp through the threshold. The obtained scaling exponents differ from the ones predicted and observed in the case in which the defects correspond to zeros in the amplitude of the global two-dimensional field. PMID- 11415051 TI - Role of higher multipoles in field-induced continuum lowering in plasmas. AB - The latest development in calculations of a continuum lowering (CL) in plasmas is based on the employment of dicenter models of the plasma state. One such theory- a percolation theory--calculated the CL defined as an absolute value of energy at which an electron becomes bound to a macroscopic portion of plasma ions (a quasi ionization). We derived analytically the value of the CL in the ionization channel, which was disregarded in the percolation theory: a quasimolecule, consisting of the two ion centers plus an electron, can become ionized in the true sense of the word before the electron would be shared by more than two ions. We derived the CL in this channel for an arbitrary ratio of charges of the two Coulomb centers, while the specific values of the CL in the percolation theory were obtained only for dicenters consisting of two identical ions. We produced our results within a purely classical approach, but proceeding from first principles. We also showed that whether the electron is bound primarily by the smaller or by the larger out of two positive charges makes a dramatic qualitative and quantitative difference for this ionization channel. This difference is revealed due to our allowance for all higher multipoles. PMID- 11415052 TI - Comment on "Phase synchronization in discrete chaotic systems". AB - Chen et al. [Phys. Rev. E 61, 2559 (2000)] recently proposed an extension of the concept of phase for discrete chaotic systems. Using the newly introduced definition of phase they studied the dynamics of coupled map lattices and compared these dynamics with phase synchronization of coupled continuous-time chaotic systems. In this paper we illustrate by two simple counterexamples that the angle variable introduced by Chen et al. fails to satisfy the basic requirements to the proper phase. Furthermore, we argue that an extension of the notion of phase synchronization to generic discrete maps is doubtful. PMID- 11415053 TI - Electrodynamic force law controversy. AB - Cavalleri et al. [Phys. Rev. E 52, 2505 (1998); Eur. J. Phys. 17, 205 (1996)] have attempted to resolve the electrodynamic force law controversy. This attempt to prove the validity of either the Ampere or Lorentz force law by theory and experiment has revealed only that the two are equivalent when predicting the force on part of a circuit due to the current in the complete circuit. However, in our analysis of internal stresses, only Ampere's force law agrees with experiment. PMID- 11415055 TI - Comment on "Algorithm for normal random numbers". AB - We have performed some simple statistical tests on the recently proposed algorithm for generating normally distributed random numbers and we find that it has some serious flaws with regard to practical application. In particular, we find that there is a statistically significant difference between the sample mean and that expected when using the original authors' recommended parameters of the algorithm. Only by significantly increasing these parameters, at the cost of considerable extra computational effort, do we get satisfactory statistics. PMID- 11415057 TI - Thermal fluctuation for the time-dependent Ginzburg-Landau simulation. AB - Thermal fluctuation for the time-dependent Ginzburg-Landau simulations is considered on the basis of the ensemble theory and the fluctuation-dissipation relation. The treatment for evaluating the thermal fluctuation for a nonconserved system is proposed and the formula for the fluctuation is presented. The magnitude of the fluctuation depends on the coarse-grained volume and the curvature of the free-energy function at thermal equilibrium. The validity of the formula is verified by numerical simulation. PMID- 11415058 TI - Dynamic structure factor of a dilute Lennard-Jones gas. AB - We calculate the leading correction term in the density expansion of the dynamic structure factor for a Lennard-Jones fluid using the Boltzmann equation. The qualitative behavior is found to be very similar to the hard-sphere result reported by Kamgar-Parsi et al. [Phys. Rev. A 35, 4781 (1987)]. A comparison was made with the results from the neutron scattering experiment by Verkerk et al. [Phys. Rev. Lett. 67, 1262 (1991)] for dilute argon gases. Several possibilities to explain the discrepancies of the present results from the experiment are proposed. PMID- 11415059 TI - Isentropes and Hugoniot curves for dense hydrogen and deuterium. AB - Multiple-shock experiments with fluid hydrogen have shown that a transition from semiconducting behavior to metal-like conductivity occurs at pressures (p) of about 140 GPa and temperatures (T) near 3000 K. We model the p-T pathway by Hugoniot curves (initial shock) and isentropes (subsequent shocks). For the calculation of these curves we apply an expression for the free energy developed recently for dense hydrogen and deuterium plasma in the regions of partial dissociation and partial ionization. Furthermore, we discuss the relations between Hugoniot curves, isentropes and the coexistence line of the plasma phase transition. PMID- 11415060 TI - Effective temperature of an aging powder. AB - The aging dynamics and the fluctuation-dissipation relation between the spontaneous diffusion induced by a random noise and the drift motion induced by a small stirring force are numerically investigated in a 3D schematic model of compacting powder: a gravity-driven lattice-gas with purely kinetic constraints. The compaction dynamics is characterized by a super-aging behavior and, in analogy with glasses, exhibits a purely dynamical time-scale-dependent effective temperature. A simple experiment to measure this quantity is suggested. PMID- 11415061 TI - First-order Freedericksz transition in the presence of light-driven feedback in nematic liquid crystals. AB - We show that feedback, which introduces a dependence of the electric field on the liquid-crystal director, renders the Freedericksz transition first order. Experimentally, the feedback is introduced as a light loop in a liquid-crystal light valve. Theoretically, we include the feedback term into the Frank free energy and we derive an amplitude equation that is valid close to the transition. The theoretical description is in qualitative agreement with the experimental observations. Depending on the values of the feedback parameters, both theory and experiment exhibit bistability, propagation of fronts, and a Maxwell point. PMID- 11415062 TI - Data clustering and noise undressing of correlation matrices. AB - We discuss an approach to data clustering. We find that maximum likelihood leads naturally to an Hamiltonian of Potts variables that depends on the correlation matrix and whose low temperature behavior describes the correlation structure of the data. For random, uncorrelated data sets no correlation structure emerges. On the other hand, for data sets with a built-in cluster structure, the method is able to detect and recover efficiently that structure. Finally we apply the method to financial time series, where the low-temperature behavior reveals a nontrivial clustering. PMID- 11415063 TI - Laplacian growth with separately controlled noise and anisotropy. AB - Conformal mapping models are used to study the competition of noise and anisotropy in Laplacian growth. For this purpose, a family of models is introduced with the noise level and directional anisotropy controlled independently. Fractalization is observed in both anisotropic growth and growth with varying noise. The fractal dimension is determined from the cluster size scaling with cluster area. For isotropic growth d=1.7, at both high and low noise. For anisotropic growth with reduced noise the dimension can be as low as d=1.5 and apparently is not universal. Also, we study the fluctuations of particle areas and observe, in agreement with previous studies, that exceptionally large particles may appear during growth, leading to pathologically irregular clusters. This difficulty is circumvented by using an acceptance window for particle areas. PMID- 11415064 TI - Markov models of non-Gaussian exponentially correlated processes and their applications. AB - We consider three different methods of generating non-Gaussian Markov processes with given probability density functions and exponential correlation functions. All models are based on stochastic differential equations. A number of analytically treatable examples are considered. The results obtained can be used in different areas such as telecommunications and neurobiology. PMID- 11415065 TI - Quenched disorder effects on deterministic inertia ratchets. AB - The effect of quenched disorder on the underdamped motion of a periodically driven particle on a ratchet potential is studied. As a consequence of disorder, current reversal and chaotic diffusion may take place on regular trajectories. On the other hand, on some chaotic trajectories disorder induces regular motion. A localization effect similar to the Golosov phenomenon sets in whenever a disorder threshold that depends on the mass of the particle is reached. Possible applications of the localization phenomenon are discussed. PMID- 11415066 TI - General pseudoadditivity of composable entropy prescribed by the existence of equilibrium. AB - The concept of composability states that entropy of the total system composed of independent subsystems is a function of entropies of the subsystems. Here, the most general pseudoadditivity rule for composable entropy is derived based only on the existence of equilibrium. PMID- 11415067 TI - Isobaric molecular dynamics simulations of hard sphere systems. AB - We describe an implementation of the Andersen algorithm for simulating the molecular dynamics in the isobaric isoenthalpic (NPH) ensemble for the hard sphere potential. The work is based on the adaptation of the Andersen algorithm to hard spheres by de Smedt et al. For a hard sphere system in the NPH ensemble, the particle velocities are not constant between collisions and we describe an efficient method for handling this part of the dynamics. The method is extended to give an NPT ensemble simulation of hard sphere systems by applying an ad hoc rescaling of the velocities. The accuracy of the algorithms is tested by comparison with traditional NVE simulation results for the structural, thermodynamic, and transport properties. PMID- 11415068 TI - Global oscillation mechanism in the stochastic Lotka model. AB - The microscopic one-parameter kinetic model of the oscillatory A+B-->2 B reaction (Lotka model) is studied using direct Monte Carlo simulations and analytical methods. Percolation is proposed as the mechanism of global oscillations that are not limited to any finite size of a system. An analytical estimate of the oscillation frequency is derived and compared to computer simulations. We also observe the transition from synchronized oscillations to specific f(-2) noise in two dimensions which was previously reported for self-organized critical models. PMID- 11415069 TI - Measurement of the percolation threshold for fully penetrable disks of different radii. AB - We perform simulations of gradient percolation to study the percolation threshold for systems of homogeneous fully penetrable disks of variable radii. We find that, if the radii follow a uniform distribution, the percolation threshold is 0.686 610+/-0.000 007. We also investigate binary dispersions, studying the influence of constitutive parameters on the percolation threshold and suggesting an empirical formula for the threshold. We find that, with the appropriate parameters, a percolation threshold of approximately 0.76 can be achieved. The minimal threshold of 0.676 339+/-0.000 004 is achieved by disks of equal radius. PMID- 11415070 TI - Kinetics of nucleation controlled formation and condensational growth of disperse particles. AB - The kinetics of nucleation controlled formation and condensational growth of disperse particles is considered under the assumptions that: (i) only a small amount of condensable substance nucleates and forms the particles that grow by condensing the rest of the substance. (ii) The condensation efficiency is a power function of the particle mass. A nontrivial perturbation theory with respect to the smallness parameter mu= (the mass of nucleated matter)/(the total mass of condensable matter) is developed allowing one to describe the source-enhanced and free (no source) condensation processes in terms of universal functions: the particle-mass spectrum and the concentration of condensable matter. The theory relies upon a scaling transformation that removes at all the smallness parameter from the evolution equations (if the nucleation rate is a power function of the concentration of condensable matter) or leaves it in the expression for the nucleation rate where this parameter defines only a concentration scale of the nucleation process (for the nucleation rates of general form). The theory is illustrated by the exact analytical solutions of the nucleation-condensation kinetic equations for three practically important cases: (i) gas-to-particle conversion in the free-molecular regime, (ii) formation and diffusion controlled condensational growth of islands on surfaces, and (iii) formation and diffusion controlled growth of disperse particles in the continuum regime. The analytical expressions for the mass spectra of growing particles are found in the case of free condensing particles. The final mass spectra in free condensing systems display rather unusual behavior: they are either singular at small particle masses or not, depending on the value of the power exponent in the mass dependence of the condensation rate. PMID- 11415071 TI - Pair distribution functions of a binary Yukawa mixture and their asymptotic behavior. AB - Based on an analytic solution of the mean spherical model for a binary hard sphere Yukawa mixture, we have examined the pair distribution functions g(ij)(r), focusing, in particular, on two aspects: (i) We present two complementary methods to compute the g(ij)(r) accurately and efficiently over the entire r range. (ii) The poles of the Laplace transforms of the pair distribution functions in the left half of the complex plane close to the origin determine the universal asymptotic behavior of the g(ij)(r). Although the meaning of the role of the subsequent poles-which typically are arranged in two branches-is not yet completely clear, there are strong indications that the distribution pattern of the poles is related to the thermodynamic state of the system. PMID- 11415072 TI - Analytical and numerical investigation of escape rate for a noise driven bath. AB - We consider a system-reservoir model where the reservoir is modulated by an external noise. Both the internal noise of the reservoir and the external noise are stationary, Gaussian, and are characterized by arbitrary decaying correlation functions. Based on a relation between the dissipation of the system and the response function of the reservoir driven by external noise, we numerically examine the model using a full bistable potential to show that one can recover the turn-over features of the usual Kramers' dynamics when the external noise modulates the reservoir rather than the system directly. We derive the generalized Kramers' rate for this nonequilibrium open system. The theoretical results are verified by numerical simulation. PMID- 11415073 TI - Multidimensional and memory effects on diffusion of a particle. AB - The diffusion of an overdamped Brownian particle in the two-dimensional (2D) channel bounded periodically by a parabola is studied, where the particle is subject to an additive white or colored noise. The diffusion rate constant D* of the particle is evaluated by the quasi-2D approximation and the effective potential approach, and the theoretical result is compared with the Langevin simulation. The properties of the diffusion rate constant are stressed for weak and strong noise cases. It is shown that, in an entropy channel, the value of D* in units of Q decreases with increasing intensity of the colored noise. In the presence of energetic barriers, a nonmonotonic behavior of the reduced diffusion rate constant D*Q-1 as a function of the noise intensity is shown. PMID- 11415074 TI - Finite-size dependence of the bridge function extracted from molecular dynamics simulations. AB - The bridge function for liquid sodium at T=373 K is obtained by using the mean spherical approximation to extrapolate the pair distribution function (PDF), calculated in molecular dynamics (MD) simulations, beyond the half simulation box length for two sizes of the MD system. The bridge function is found to strongly depend on the total number of particles used in the simulation cell. This dependency leads to a spurious maximum of the static structure factor at long wavelengths, obtained from the reference hypernetted-chain approximation (RHNC) with the MD system used as a reference system (RHNC-MD). A simple self-consistent procedure, proposed to account for the finite-size effects in the bridge function, allows one to efficiently correct the RHNC-MD static structure factor for all unphysical manifestations. PMID- 11415075 TI - Thermodynamic pressure in nonlinear nonequilibrium thermodynamics of dilute nonviscous gases. AB - In this paper, using extended thermodynamics, we build up a nonlinear theory for a dilute nonviscous gas under heat flux. The fundamental fields are the density, the velocity, the internal energy density, and the heat flux. The constitutive theory is builtup without approximations. We single out the nonlinear complete expressions of the Gibbs equation and of the nonequilibrium pressure. In particular, we determine the complete expressions furnished by the theory for the nonequilibrium pressure tensor and thermodynamic pressure, i.e., the derivative of the nonequilibrium internal specific entropy with respect to the specific volume, times the nonequilibrium temperature. In a second-order approximation these expressions are identical with those obtained in Phys. Rev. E 51, 158 (1995), using information theory. PMID- 11415076 TI - Application of hard sphere perturbation theory for thermodynamics of model liquid metals. AB - Hard sphere perturbation theory (HSPT) has contributed toward the fundamental understanding of dense fluids for over 30 years. In recent decades, other techniques have been more popular. In this paper, we argue for the revival of hard sphere perturbation theory for the study of thermodynamics of dense liquid in general, and in liquid metal in particular. The weakness of HSPT is now well understood, and can be easily overcome by using a simple convenient Monte Carlo method to calculate the intrinsic error of HSPT free energy density. To demonstrate this approach, we consider models of liquid aluminum and sodium. We obtain the intrinsic error of HSPT with the Monte Carlo method. HSPT is shown to provide a lower free energy upper bound than one-component plasma (OCP) for alkali metals and polyvalent metals. We are thus able to provide insight into the long standing observation that a OCP is a better reference system than a HS for alkali metals. PMID- 11415077 TI - Mean-field treatment of polarons in strong electrolytes. AB - Using variational estimates for the grand partition function, we have developed a microscopic theory of an excess electron in an ionic liquid at high ion concentrations. We have derived the free-energy functional for the electron and have calculated electron energies for the ground and the first excited states as well as electron-ion correlation functions versus thermodynamic parameters of liquid and parameters of electron-ion potentials. We have found that the energetic characteristics of solvated electron are mainly determined by the Coulomb interaction which gives birth to polaronlike states, while ion cores have a pronounced quantitative effect on these characteristics. The local solvent structure around the excess electron is determined by the mean field induced by ions. Using the method developed we have calculated polaron characteristics in molten salts, such as the maximum of the absorption spectrum and its variations caused by changes in temperature, density, and composition of the electrolyte. The data obtained are in agreement with experiments and computer simulations. PMID- 11415078 TI - Effect of helicity on the effective diffusivity for incompressible random flows. AB - The advection of a passive scalar by a quenched (frozen) incompressible velocity field is studied by extensive high precision numerical simulation and various approximation schemes. We show that second-order self-consistent perturbation theory, in the absence of helicity, perfectly predicts the effective diffusivity of a tracer particle in such a field. In the presence of helicity in the flow, simulations reveal an unexpectedly strong enhancement of the effective diffusivity which is highly nonperturbative and most visible when the bare molecular diffusivity of the particle is small. We develop and analyze a series of approximation schemes which indicate that this enhancement of the diffusivity is due to a second order effect, whereby the helical component of the field, which does not directly renormalize the effective diffusivity, enhances the strength of the nonhelical part of the flow, which in turn renormalizes the molecular diffusivity. We show that this renormalization is most important at a low bare molecular diffusivity, in agreement with numerical simulations. PMID- 11415079 TI - Reformulation of density functional theory for generation of the nonuniform density distribution. AB - The concept of universality of the free energy density functional and the weighted density approximation are combined to provide the density distribution profile of nonuniform fluids from the predictions of integral equation theory for the corresponding uniform fluids. To obtain the expression for the free energy as a function of the density distribution, the present formalism expresses the difference of the first order direct correlation function of a nonuniform fluid with respect to its uniform fluid counterpart as a function of the weighted density, which is also a function of the space position. The input parameters used in the present approach are the radial distribution function and the second order direct correlation function of the corresponding uniform fluid. All of these parameters can be easily obtained from numerical solution of Ornstein Zernike integral equation theory. The present approach is based on the formalism of classical density functional theory (DFT) and its application to two kinds of fluid under different external potentials is presented. The agreement of the theoretical predictions with the corresponding computer simulation data is good. The present formulation of DFT can treat fluids of different interaction potential under nonzero external fields in a unified way. PMID- 11415080 TI - Dynamic scaling theory for a tethered membrane in solution. AB - We present the dynamic scaling behavior for the specific viscosity and diffusion coefficient of a single membrane and membranes with nonzero concentration in solution. Starting from the membrane free energies, we derive their Langevin equations. The corresponding Kirkwood diffusion equation, describing the time evolution in configuration space, contains two kinds of time scales that are separated by the external dimension 4/(2-D) where D is the dimension of the internal space. These time scale separation behaviors depend strongly on the hydrodynamic screening effect. For a single membrane solution, we resolve the dynamic scaling exponents for the diffusion coefficient and intrinsic viscosity by the dimension reduction method. For a concentrated membrane solution, the effective excluded volume strength and draining parameter are introduced. The effective medium argument is applied to obtain a concentration dependent power law form for the specific viscosity and diffusion coefficient, whose results contribute to a fundamental understanding of membrane in solution and of hydrodynamic screening and excluded volume effects in many different solvents. PMID- 11415081 TI - Forced two-dimensional turbulence in spectral and physical space. AB - Two-dimensional (2D) turbulence in the energy range exhibits nonuniversal features, manifested in the departure (at low k) from the k(-5/3) energy spectrum law, variable energy flux, and irregular, nonlocal transfers. To unravel the underlying mechanism we conducted a detailed study of the 2D turbulence in spectral and physical space. It revealed complex multiscale organization of vorticity field and dynamic processes, ranging from large-scale meandering jets to strong localized vortices. The latter bear prime responsibility for the nonuniversal behavior of 2D turbulence, and we examined their statistical features and the growth mechanism. Our results are based on the numeric simulation of 2D turbulence on the 512 grid under different forcing-dissipation conditions. PMID- 11415082 TI - Light-scattering spectra of supercooled molecular liquids. AB - The light-scattering spectra of molecular liquids are derived within a generalized hydrodynamics. The wave-vector and scattering-angle dependencies are given in the most general case and the change of the spectral features from liquid to solidlike is discussed without phenomenological model assumptions for (general) dielectric systems without long-ranged order. Exact microscopic expressions are derived for the frequency dependent transport kernels, generalized thermodynamic derivatives, and the background spectra. PMID- 11415083 TI - Dynamics in a supercooled molecular liquid: theory and simulations. AB - We report extensive simulations of liquid supercooled states for a simple three site molecular model, introduced by Lewis and Wahnstrom [Phys. Rev. E 50, 3865 (1994)] to mimic the behavior of orthoterphenyl. The large system size and the long simulation length allow us to calculate very precisely (in a large q-vector range) self-correlation and collective correlation functions, providing a clean and simple reference model for theoretical descriptions of molecular liquids in supercooled states. The time and wave-vector dependence of the site-site correlation functions are compared (neglecting the molecular constraints) with detailed ideal mode-coupling theory predictions. Except for the wave-vector region where the dynamics are controlled by the center of mass (around 9 nm(-1)), the theoretical predictions compare very well with the simulation data. PMID- 11415084 TI - Model for the atomic-scale structure of the homogeneous cooling state of granular fluids. AB - It is proposed that the equilibrium generalized mean spherical model of fluid structure may be extended to nonequilibrium states with equation of state information used in equilibrium replaced by an exact condition on the two-body distribution function. The model is applied to the homogeneous cooling state of granular fluids, and upon comparison to molecular-dynamics simulations, is found to provide an accurate picture of the pair distribution function. PMID- 11415085 TI - Modified lattice Boltzmann method for compressible fluid simulations. AB - A modified lattice Boltzmann algorithm is shown to have much better stability to growing temperature perturbations, when compared with the standard lattice Boltzmann algorithm. The damping rates of long-wavelength waves, which determine stability, are derived using a collisional equilibrium distribution function which has the property that the Euler equations are obtained exactly in the limit of zero time step. Using this equilibrium distribution function, we show that our algorithm has inherent positive hyperviscosity and hyperdiffusivity, for very small values of viscosity and thermal diffusivity, which are lacking in the standard algorithm. Short-wavelength modes are shown to be stable for temperatures greater than a lower limit. Results from a computer code are used to compare these algorithms, and to confirm the damping rate predictions made analytically. Finite amplitude sound waves in the simulated fluid steepen, as expected from gas dynamic theory. PMID- 11415086 TI - Linear response of vibrated granular systems to sudden changes in the vibration intensity. AB - The short-term memory effects recently observed in vibration-induced compaction of granular materials are studied. It is shown that they can be explained by means of quite plausible hypothesis about the mesoscopic description of the evolution of the system. The existence of a critical time separating regimes of "anomalous" and "normal" responses is predicted. A simple model fitting into the general framework is analyzed in the detail. The relationship between this paper and previous studies is discussed. PMID- 11415087 TI - Convection in a fully immersed granular slurry. AB - Using vertical vibrations, we have induced convection in a granular system completely immersed in fluid inside a cylindrical container. Convection is established in a single convection roll, which carries the material upward in the center and downward along the side walls of the container. We measure the rise times of tracer particles embedded in the medium at various depths along the central axis of the system. By fitting this data to an appropriate functional form, we obtain information on the depth dependence of convection velocity. Significant differences are found in the frequency and acceleration amplitude dependence of convection between wet and otherwise identical dry systems. In addition, we find that a power-law form provides a better fit to our data than the logarithmic form used in dry systems. PMID- 11415088 TI - Influence of shape on ordering of granular systems in two dimensions. AB - We investigate ordering properties of two-dimensional granular materials using several shapes created by welding ball bearings together. Ordered domains form much more easily in two than in three dimensions, even when configurations lack long-range order. The onset of ordered domains occurs near a packing density of 0.8, a phenomenon observed previously for disks. One of our shapes, the trapezoid, has packings that remain disordered and near the transition density even after annealing by shaking. Although random packings are unstable for disks and many other shapes in two dimensions, trapezoid packings provide an approach to studying two-dimensional randomness. We also find that the rotational symmetry of a shape is an excellent predictor of how easily it orders, and a potential guide to identifying two-dimensional shapes that remain random after annealing. PMID- 11415089 TI - Bifurcation diagram for compartmentalized granular gases. AB - The bifurcation diagram for a vibrofluidized granular gas in N connected compartments is constructed and discussed. At vigorous driving, the uniform distribution (in which the gas is equi-partitioned over the compartments) is stable. But when the driving intensity is decreased this uniform distribution becomes unstable and gives way to a clustered state. For the simplest case, N=2, this transition takes place via a pitchfork bifurcation but for all N>2 the transition involves saddle-node bifurcations. The associated hysteresis becomes more and more pronounced for growing N. In the bifurcation diagram, apart from the uniform and the one-peaked distributions, also a number of multipeaked solutions occur. These are transient states. Their physical relevance is discussed in the context of a stability analysis. PMID- 11415090 TI - Hydrodynamics of an open vibrated granular system. AB - Using the hydrodynamic description and molecular dynamics simulations, the steady state of a fluidized granular system in the presence of gravity is studied. For an open system, the density profile exhibits a maximum, while the temperature profile goes through a minimum at high altitude, beyond that the temperature increases with the height. The existence of the minimum is explained by the hydrodynamic equations if the presence of a collisionless boundary layer is taken into account. The energy dissipated by interparticle collisions is also computed. A good agreement is found between theory and simulation. The relationship with previous works is discussed. PMID- 11415091 TI - Lyapunov spectrum of granular gases. AB - We calculate and study the Lyapunov spectrum of a granular gas maintained in a steady state by an isokinetic thermostat. Considering restitution coefficients greater than unity allows us to show that the spectra change smoothly and continuously at equilibrium. The shearing instability of the granular gas, however, provokes an abrupt change in the structure of the spectrum. The relationship between various physically relevant quantities and the energy dissipation rate differs from previously studied nonequilibrium steady states. PMID- 11415092 TI - Grain reconstruction of porous media: application to a low-porosity Fontainebleau sandstone. AB - The fundamental issue of reconstructing a porous medium is examined anew in this paper, thanks to a sample of low-porosity Fontainebleau sandstone that has been analyzed by computed microtomography. Various geometric properties are determined on the experimental sample. A statistical property, namely, the probability density of the covering radius, is determined. This is used in order to reconstruct a porous medium by means of a Poissonian generation of polydisperse spheres. In a second part, the properties of the real experimental sample and of the reconstructed one are compared. The most important success of the present reconstruction technique is the fact that the numerical sample percolates despite its low porosity. Moreover, other geometrical features and conductivity are found to be in good agreement. PMID- 11415093 TI - Inelastic collapse in simple shear flow of a granular medium. AB - Previous investigations have shown that inelastic collapse is a common feature of inelastic, hard-sphere simulations of nondriven (or unforced) flows, provided that the coefficient of restitution is small enough. The focus of the current effort is on a driven system, namely, simple shear flow. Two-dimensional, hard sphere simulations have been carried out over a considerable range of restitution coefficients (r), solids fractions (nu), and numbers of particles (N). The results indicate that inelastic collapse is an integral feature of the sheared system. Similar to nondriven systems, this phenomenon is characterized by a string of particles engaging in numerous, repeated collisions just prior to collapse. The collapsed string is typically oriented along a 135 degrees angle from the streamwise direction. Inelastic collapse is also found to be more likely in systems with lower r, higher nu, and higher N, as is true for unforced systems. Nonetheless, an examination of the boundary between the collapsed and noncollapsed states reveals that the sheared system is generally more "resistant" to inelastic collapse than its nondriven counterpart. Furthermore, a dimensionless number V* is identified that represents the magnitude of the initial fluctuating velocities relative to that of a characteristic steady-state velocity (i.e., the product of shear rate and particle diameter). For values of V*>>O(1), the transient portion of the simulation is found to be more reminiscent of a nondriven system (i.e., isotropic particle bunching is observed instead of diagonal particle bands). PMID- 11415094 TI - Evidence for universality within the classes of deterministic and stochastic sandpile models. AB - Recent numerical studies have provided evidence that within the family of conservative, undirected sandpile models with short range dynamic rules, deterministic models such as the Bak-Tang-Wiesenfeld model [P. Bak, C. Tang, and K. Wiesenfeld, Phys. Rev. Lett. 59, 381 (1987)] and stochastic models such as the Manna model [S. S. Manna, J. Phys. A 24, L363 (1991)] belong to different universality classes. In this paper we examine the universality within each of the two classes in two dimensions by numerical simulations. To this end we consider additional deterministic and stochastic models and use an extended set of critical exponents, scaling functions, and geometrical features. Universal behavior is found within the class of deterministic Abelian models, as well as within the class of stochastic models (which includes both Abelian and non Abelian models). In addition, it is observed that deterministic but non-Abelian models exhibit critical exponents that depend on a parameter, namely they are nonuniversal. PMID- 11415095 TI - Effects of gravity and nonlinearity on the waves in the granular chain. AB - The solitary signal observed in a horizontal granular chain changes its speed and form due to gravity in a vertical chain. We find that all the propagating signals in a vertical chain follow power laws in depth for propagating speed, grain velocity, amplitude, and width. This stems from the power-law type changing of elastic properties in a medium under gravity. The propagation may be separated into two types according to the behavior of the power-law exponents, depending on the strength of the nonlinearity. We show that the power-law exponents are constants in the strength of the impulse in the weakly nonlinear regime, while they depend on the strength of the impulse in the strongly nonlinear regime. We derive power-law exponents for the weakly nonlinear regime analytically and try to understand the behaviors of the strongly nonlinear regime through analytical treatment. PMID- 11415096 TI - Granular temperature profiles in three-dimensional vibrofluidized granular beds. AB - The motion of grains in a three-dimensional vibrofluidized granular bed has been measured using the technique of positron emission particle tracking, to provide three-dimensional packing fraction and granular temperature distributions. The mean square fluctuation velocity about the mean was calculated through analysis of the short time mean squared displacement behavior, allowing measurement of the granular temperature at packing fractions of up to eta approximately 0.15. The scaling relationship between the granular temperature, the number of layers of grains, and the base velocity was determined. Deviations between the observed scaling exponents and those predicted by recent theories are attributed to the influence of dissipative grain-sidewall collisions. PMID- 11415097 TI - Self-organized criticality in a sheared granular stick-slip system. AB - We present an analysis of results obtained from a mechanical apparatus consisting of an annular plate shearing over a granular bed. The size, energy dissipation, and duration of slips in the system exhibit power-law distributions and a 1/f(2) power spectrum, in accordance with self-organized criticality. We draw similarities with earthquakes. PMID- 11415098 TI - Flow dichroism in critical colloidal fluids. AB - Due to long-range correlations and slow dynamics of concentration fluctuations in the vicinity of the gas-liquid critical point, shear flow is very effective in distorting the microstructure of near-critical fluids. The anisotropic nature of the shear-field renders the microstructure highly anisotropic, leading to dichroism. Experiments on the dichroic behavior can thus be used to test theoretical predictions on microstructural order under shear flow conditions. We performed both static and dynamic dichroism and turbidity measurements on a colloid-polymer mixture, existing of silica spheres (radius 51 nm) and polydimethylsiloxane polymer (molar weight 204 kg/mol). Sufficiently far away from the critical point, in the mean-field region, the experimental data are in good agreement with theory. Very close to the critical point, beyond mean field, for which no theory exists yet, an unexpected decrease of dichroism on approach of the critical point is observed. Moreover, we do not observe critical slowing down of shear-induced dichroism, right up to the critical point, in contrast to the turbidity. PMID- 11415099 TI - Topology of slightly polydisperse real foams. AB - The topology of slightly polydisperse, (meta-)stable, real foams was investigated by means of optical tomography associated with a numerical reconstruction procedure. The values of the mean numbers of faces per bubble and edges per face were very close to Matzke's data (1946). The real foams were essentially disordered and possessed a noncentered symmetry, and ideal structures also could not be observed. The disorder was quantified by the second moment of the edge per face and face per bubble distributions, and also by a statistical correlation coefficient between the numbers of edges of adjacent faces. It was found that the edge distributions of the internal bubbles, and not of the external ones, were significantly anticorrelated even during foam aging, which provided a measure of the disorder in the foam. No obvious relationship could be deduced between the isoperimetric quotient and the face combination in an individual bubble. Eventually, it was shown that the physical boundaries of the foam sample had no influence on the foam topology beyond a single bubble layer. PMID- 11415100 TI - Shear viscosity of phase-separating polymer blends with viscous asymmetry. AB - Rheo-optical measurements of phase separating polymer mixtures under simple shear flow have been used to investigate the influence of domain morphology on the viscosity of emulsionlike polymer blends, in which the morphology under weak shear is droplets of one coexisting phase dispersed in a matrix of the second. The structure and viscosity of low-molecular-weight polybutadiene and polyisoprene mixtures, phase separated by quenching to a temperature inside the coexistence region of the phase diagram, were measured as a function of shear rate and composition. In the weak shear regime, the data are in qualitative agreement with an effective medium model for non-dilute suspensions of slightly deformed interacting droplets. In the strong shear regime, where a stringlike pattern appears en route to a shear-homogenized state, the data are in qualitative agreement with a simple model that accounts for viscous asymmetry in the components. PMID- 11415101 TI - Diffusing-wave spectroscopy of nonergodic media. AB - We introduce an elegant method that allows the application of diffusing-wave spectroscopy (DWS) to nonergodic, solidlike samples. The method is based on the idea that light transmitted through a sandwich of two turbid cells can be considered ergodic even though only the second cell is ergodic. If absorption and/or leakage of light take place at the interface between the cells, we establish a so-called "multiplication rule," which relates the intensity autocorrelation function of light transmitted through the double-cell sandwich to the autocorrelation functions of individual cells by a simple multiplication. To test the proposed method, we perform a series of DWS experiments using colloidal gels as model nonergodic media. Our experimental data are consistent with the theoretical predictions, allowing quantitative characterization of nonergodic media and demonstrating the validity of the proposed technique. PMID- 11415102 TI - Critical behavior of gelation probed by the dynamics of latex spheres. AB - We report a quasielastic light scattering study of the dynamics of large latex probe particles (R=225 nm) in gelatin solution undergoing gelation. We show that by focusing on the short-time and long-time behavior of the autocorrelation function, it is possible to simply interpret out data in terms of the divergence of the viscosity and emergence of the shear elastic modulus near the gel point. Our crude analysis allows us to grasp the critical behavior of gelation and to obtain the two critical exponents of the transport properties. PMID- 11415103 TI - Droplet polydispersity and shape fluctuations in AOT [bis(2 ethylhexyl)sulfosuccinate sodium salt] microemulsions studied by contrast variation small-angle neutron scattering. AB - Microemulsions consisting of AOT water, and decane or iso-octane are studied in the region of the phase diagram where surfactant covered water droplets are formed. The polydispersity and shape fluctuations of the microemulsion droplets are determined and compared in the two different alkane types. Conductivity measurements show that there is a pronounced dependence of the temperature behavior of the microemulsion on the type of alkane used. In both cases the microemulsion droplets start to form larger aggregates when the temperature increases. But in the system with decane this aggregation temperature occurs at a temperature about 10 degrees C lower than in a similar system with iso-octane. Aggregation phenomena are avoided and the two systems are at approximately the same reduced temperature with respect to the aggregation temperature when the temperature of the AOT/D(2)O/decane microemulsion is 10 degrees C and the temperature of the AOT/D(2)O/iso-octane microemulsion is 20 degrees C. Contrast variation small-angle neutron scattering measurements are performed at these temperatures on systems with volume fractions of 5% D2O+AOT by varying the scattering length density of the alkane. The small-angle scattering for 11 different contrasts evenly distributed around the match points are studied for each sample. The scattering data for the different contrasts are analyzed using a molecular constrained model for ellipsoidal droplets of water covered by AOT, interacting as polydisperse hard spheres. All contrasts are fitted simultaneously by taking the different contrast factors into account. The analysis show that at the same reduced temperature with respect to the aggregation temperature the droplet size, polydispersity index, the size of the shape fluctuations are similar in the two systems. A polydispersity index (sigma/R of the Gaussian size distribution) of 16% and an average axis ratio of the droplets of 1.56 is found in the AOT/D(2)O/decane microemulsion. In the AOT/D(2)O/iso-octane system the polydispersity index is also 16% while the axis ratio is 1.72. The bending elastic constant kappa and the Gaussian bending elastic constant kappa; can be estimated from these numbers. For AOT/D(2)O/decane we find kappa=3.4 k(B)T and kappa=-5.9 k(B)T and for AOT/D(2)O/iso-octane we find kappa=2.35k(B)T and kappa= 3.8k(B)T, where k(B) is the Boltzmann constant and T is the absolute temperature. PMID- 11415104 TI - Liquid-liquid equilibrium for monodisperse spherical particles. AB - A system of identical particles interacting through an isotropic potential that allows for two preferred interparticle distances is numerically studied. When the parameters of the interaction potential are adequately chosen, the system exhibits coexistence between two different liquid phases (in addition to the usual liquid-gas coexistence). It is shown that this coexistence can occur at equilibrium, namely, in the region in which the liquid is thermodynamically stable. PMID- 11415105 TI - Glass transition and density fluctuations in the fragile glass former orthoterphenyl. AB - High-resolution Brillouin light scattering is used to measure the dynamic structure factor of the fragile glass former orthoterphenyl (OTP) in a wide temperature range around the glass transition region and up to the boiling point. The whole set of spectra is described in terms of a phenomenological generalized hydrodynamic model. In the supercooled phase, we show the contemporary existence of the structural process, whose main features come out to be consistent with the results obtained with other spectroscopies, and of a secondary, activated process, which occurs on the 10(-11) s time scale and has a low activation energy (E(f)(a)=0.28 kcal/mol). This latter process, which is also present in the glassy phase and seems to be insensitive to the glass transition, is attributed to the coupling between the density modes and intramolecular degrees of freedom. In the normal liquid phase, the two processes merge together, and the resulting characteristic time is no longer consistent with those derived with other spectroscopies. The analysis points to the conclusion that, for what concerns the long-wavelength density fluctuations in fragile glass formers such as OTP, the universal dynamical features related to the glass transition come out clearly only in the supercooled phase and at frequencies lower than approximately 10(6) Hz. PMID- 11415106 TI - Unusual features of long-range density fluctuations in glass-forming organic liquids: a Rayleigh and Rayleigh-Brillouin light scattering study. AB - A new feature of glass-forming liquids, i.e., long-range density fluctuations of the order of 100 nm, has been extensively characterized by means of static light scattering, photon correlation spectroscopy and Rayleigh-Brillouin spectroscopy in orthoterphenyl (OTP) and 1,1-di(4(')-methoxy-5(')methyl-phenyl)-cyclohexane (BMMPC). These long-range density fluctuations result in the following unusual features observed in a light scattering experiment, which are not described by the existing theories: (i) strong q-dependent isotropic excess Rayleigh intensity, (ii) additional slow component in the polarized photon correlation function, and (iii) high Landau-Placzek ratio. These unusual features are equilibrium properties of the glass-forming liquids and depend only on temperature, provided that the sample has been equilibrated long enough. The temperature-dependent equilibration times were measured for BMMPC and are about 11 orders of magnitude longer than the alpha process. It was found that the glass forming liquid OTP may occur in two states: with and without long-range density fluctuations ("clusters"). We have characterized the two states by static and dynamic light scattering in the temperature range from T(g) to T(g)+200 K. The relaxation times of the alpha process as well as the parameters of the Brillouin line are identical in both OTP with and without clusters. The alpha process (density fluctuations) in OTP was characterized by measuring either the polarized (VV) or depolarized (VH) correlation function, which are practically identical and q-independent. This feature, which is commonly observed in glass-forming liquids, is not fully explained by the existing theories. PMID- 11415107 TI - Molecular dynamics studies of strongly coupled charged particle bilayers at finite temperatures. AB - The structure of strongly coupled charged particle bilayers was investigated using molecular dynamics simulation, in a wide range of the plasma coupling parameter Gamma=20-100. The simulations showed the existence of a series of structural transformations, controlled by the separation of the layers. At high values of Gamma a pronounced long-range order was found to develop at intermediate layer separations, with staggered square lattice configuration. The results show a fair agreement with those obtained by hypernetted chain calculations, in terms of intralayer and interlayer pair correlation functions and structure functions. PMID- 11415108 TI - Velocity correlations and diffusion during sedimentation. AB - We study the dynamics of sedimenting non-Brownian particles under steady-state conditions in two-dimensional geometry. We concentrate on the autocorrelation functions of the velocity fluctuations and the corresponding memory functions and diffusion coefficients as functions of Phi(V) for small but finite Reynolds numbers. For the numerical simulations we have chosen the model of Schwarzer [Phys. Rev. E 52, 6461 (1995)] where a continuum liquid phase is coupled through Stokesian friction to a discrete particle phase with volume fraction Phi(V). We find that the steady-state velocity fluctuations are spatially highly anisotropic and the correlation functions parallel to gravity have nonexponential time dependence similar to that of purely dissipative systems with strong interactions. The corresponding memory functions also show nontrivial behavior. Diffusion along the direction of gravity is much faster than perpendicular to it, with the anisotropy decreasing as either the Reynolds number or the volume fraction increases. PMID- 11415109 TI - Interface fluctuations, bulk fluctuations, and dimensionality in the off equilibrium response of coarsening systems. AB - The relationship between statics and dynamics proposed by Franz, Mezard, Parisi, and Peliti (FMPP) for slowly relaxing systems [Phys. Rev. Lett. 81, 1758 (1998)] is investigated in the framework of nondisordered coarsening systems. Separating the bulk from interface response we find that for statics to be retrievable from dynamics the interface contribution must be asymptotically negligible. How fast this happens depends on dimensionality. There exists a critical dimensionality above that the interface response vanishes like the interface density and below that it vanishes more slowly. At d=1 the interface response does not vanish leading to the violation of the FMPP scheme. This behavior is explained in terms of the competition between curvature-driven and field-driven interface motion. PMID- 11415110 TI - Dynamical susceptibility of solid ferrocolloids. AB - A theoretical analysis of the dynamical magnetic susceptibility of a frozen nondilute ferrocolloid is presented. The steric and dipole-dipole interactions between ferroparticles are taken into account. Two systems are studied. The first is the homogeneous solid ferrocolloid with separate individual particles. The second is a ferrocolloid with heterogeneous chainlike aggregates. The effect of the magnetodipole interparticle interaction on the real and imaginary parts of the dynamical susceptibilities of these systems is estimated. PMID- 11415111 TI - Shape oscillations of a viscoelastic drop. AB - Small-amplitude axisymmetric shape deformations of a viscoelastic liquid drop in microgravity are theoretically analyzed. Using the Jeffreys constitutive equation for linear viscoelasticity, the characteristic equation for the frequency and decay factor of the shape oscillations is derived. Asymptotic analysis of this equation is performed in the low- and high-viscosity limits and for the cases of small, moderate, and large elasticities. Elastic effects are shown to give rise to a type of shape oscillation that does not depend on the surface tension. The existence of such oscillations is confirmed by numerical solution of the characteristic equation in various regimes. A method for determining the viscoelastic properties of highly viscous liquids based upon experimental measurements of the frequency and damping rate of such shape oscillations is suggested. PMID- 11415112 TI - Low-temperature behavior of core-softened models: water and silica behavior. AB - A core-softened model of a glass forming fluid is numerically studied in the limit of very low temperatures. The model shows two qualitatively different behaviors depending on the strength of the attraction between particles. For no or low attraction, the changes of density as a function of pressure are smooth, although hysteretic due to mechanical metastabilities. For larger attraction, sudden changes of density upon compressing and decompressing occur. This global mechanical instability is correlated to the existence of a thermodynamic first order amorphous-amorphous transition. The two different behaviors obtained correspond qualitatively to the different phenomenology observed in silica and water. PMID- 11415113 TI - Phase-field simulations of dendritic crystal growth in a forced flow. AB - Convective effects on free dendritic crystal growth into a supercooled melt in two dimensions are investigated using the phase-field method. The phase-field model incorporates both melt convection and thermal noise. A multigrid method is used to solve the conservation equations for flow. To fully resolve the diffuse interface region and the interactions of dendritic growth with flow, both the phase-field and flow equations are solved on a highly refined grid where up to 2.1 million control volumes are employed. A multiple time-step algorithm is developed that uses a large time step for the flow-field calculations while reserving a fine time step for the phase-field evolution. The operating state (velocity and shape) of a dendrite tip in a uniform axial flow is found to be in quantitative agreement with the prediction of the Oseen-Ivantsov transport theory if a tip radius based on a parabolic fit is used. Furthermore, using this parabolic tip radius, the ratio of the selection parameters without and with flow is shown to be close to unity, which is in agreement with linearized solvability theory for the ranges of the parameters considered. Dendritic sidebranching in a forced flow is also quantitatively studied. Compared to a dendrite growing at the same supercooling in a diffusive environment, convection is found to increase the amplitude and frequency of the sidebranches. The phase-field results for the scaled sidebranch amplitude and wavelength variations with distance from the tip are compared to linear Wentzel-Kramers-Brillouin theory. It is also shown that the asymmetric sidebranch growth on the upstream and downstream sides of a dendrite arm growing at an angle with respect to the flow can be explained by the differences in the mean shapes of the two sides of the arm. PMID- 11415114 TI - Topography and instability of monolayers near domain boundaries. AB - We theoretically study the topography of a biphasic surfactant monolayer in the vicinity of domain boundaries. The differing elastic properties of the two phases generally lead to a nonflat topography of "mesas," where domains of one phase are elevated with respect to the other phase. The mesas are steep but low, having heights of up to 10 nm. As the monolayer is laterally compressed, the mesas develop overhangs and eventually become unstable at a surface tension of about K(deltac(0))(2) (deltac(0) being the difference in spontaneous curvature and K a bending modulus). In addition, the boundary is found to undergo a topography induced rippling instability upon compression, if its line tension is smaller than about Kdeltac(0). The effect of diffuse boundaries on these features and the topographic behavior near a critical point are also examined. We discuss the relevance of our findings to several experimental observations related to surfactant monolayers: (i) small topographic features recently found near domain boundaries; (ii) folding behavior observed in mixed phospholipid monolayers and model lung surfactants; (iii) roughening of domain boundaries seen under lateral compression; (iv) the absence of biphasic structures in tensionless surfactant films. PMID- 11415115 TI - Temporal evolution of relaxation in rubbed polystyrene thin films. AB - Temporal relaxation of rubbed polystyrene (PS) thin films (50 nm thick) was studied by probing the decay of the optical anisotropy in the sample. Our results show that the relaxation process is characterized by two single-exponential decays plus a temperature-dependent constant. Both relaxations are orders of magnitude faster than the main chain (alpha) relaxation at temperatures well below (>10 degrees C)T(g), with activation energies 3.0 kcal/mol and 5.1 kcal/mol, respectively, either of which are much smaller than that of the alpha relaxation ( approximately 50 kcal/mol). The decay time constants are found to be independent of the sample molecular weights M(w) (=13.7 K to 550 K Daltons, M(w)/M(n)-1/2, a crossover regime leads to an systematic overestimate of the Hurst exponent. PMID- 11415148 TI - Drift velocity of C(60)(+) in gases follows rarefied gas dynamics. AB - The drift velocities of C(60)(+) in He, Ne, Ar, and Kr were found to be estimated with high accuracy using the drag coefficient of a solid body in free molecule flow. That is, massive C(60)(+) behaves in gases as if it were a large classical body. PMID- 11415149 TI - Test of the fractional Debye-Stokes-Einstein equation in low-molecular-weight glass-forming liquids under condition of high compression. AB - From temperature studies at ambient pressure, it was pointed out for several glass-forming liquids that the alpha-relaxation time (tau) can be related to the dc-ionic conductivity (sigma) through the phenomenological fractional Debye Stokes-Einstein (DSE) equation. In the present paper we test the validity of fractional DSE equation for relaxation data obtained from pressure variable experiments. To this end we carried out broadband dielectric measurements (10 mHz 10 MHz) in a wide range of pressures (0.1-300 MPa). The material under study were N,N-diglycidyl-4-glycidyloxyaniline and N,N-diglycidylaniline. As a result we found that the fractional DSE equation is also obeyed for pressure pathways. PMID- 11415150 TI - Effect of the anisotropy of the cells on the topological properties of two- and three-dimensional froths. AB - We study the effect of the anisotropy of the cells on the topological properties of monodisperse two- and three-dimensional (3D) froths. These froths are built by Voronoi tessellation of actual assemblies of monosize disks (2D) and of many numerical packings of monosize disks (2D) and spheres (3D). We show that some topological properties of these froths can be simply related to the anisotropy of the cells. PMID- 11415151 TI - (2+1)-dimensional stochastic growth model and its application to some experimental observations. AB - A (2+1)-dimensional stochastic growth model is proposed in order to understand various experimental observations at low temperature growth. The model includes step attached adatom hopping in the upward direction to provide a source of additional particle current. The flexibility obtained due to additional current control allows one to express most of the experimental observations within the framework of this model. It is argued that the time evolution exponent beta for surface roughness is >0.5 only if upward hops are included. Simulations using this model are shown to be qualitatively consistent with the observations for homoepitaxial growth on Cu(100) and Ge(100) at different temperatures. PMID- 11415152 TI - Observation of a pretransitional effect near a virtual smectic-A--smectic-C* transition. AB - Unusual softening of the layer compression modulus B has been observed near the phase boundary where the smectic-C* phase vanishes in a naphtalene-based liquid crystal mixture. From the systematic study of x-ray and layer compression measurements, this unusual effect is attributed to the pretransitional softening near a virtual smectic-A-smectic-C* phase transition in the smectic-A phase, which no longer appears on the thermoequilibrium phase diagram. This phenomenon is similar but not equivalent to supercritical behavior. PMID- 11415153 TI - Cluster Monte Carlo simulations of the nematic-isotropic transition. AB - We report the results of simulations of the three-dimensional Lebwohl-Lasher model of the nematic-isotropic transition using a single cluster Monte Carlo algorithm. The algorithm, first introduced by Kunz and Zumbach to study two dimensional nematics, is a modification of the Wolff algorithm for spin systems, and greatly reduces critical slowing down. We calculate the free energy in the neighborhood of the transition for systems up to linear size 70. We find a double well structure with a barrier that grows with increasing system size. We thus obtain an upper estimate of the value of the transition temperature in the thermodynamic limit. PMID- 11415154 TI - Velocity of an electric-field-induced synclinic solitary wave invading the anticlinic liquid crystal phase. AB - The electric-field dependence of the velocity of synclinic fingers invading the anticlinic phase is determined by a time-of-flight technique. The time delay for a rapid increase in the transmitted optical intensity through the sample is measured between two points as a function of their separation along the trajectory of the solitary wave. The data are quantitatively consistent with the rapid velocities deduced from a previous measurement [Liq. Cryst. 27, 249 (2000)], demonstrating that the previous data were not affected by multiple nucleation sites occurring at higher fields. PMID- 11415155 TI - Comment on "1/f noise in the Bak-Sneppen model". AB - Contrary to the recently published results by Daerden and Vanderzande [Phys. Rev. E 53, 4723 (1996)], we show that the time correlation function in the random neighbor version of the Bak-Sneppen model can be well approximated by an exponential giving rise to a 1/f(2) power spectrum. PMID- 11415156 TI - Mode-coupling theory of the fifth-order Raman spectrum of an atomic liquid. AB - A fully microscopic molecular hydrodynamic theory for the two-dimensional (fifth order) Raman spectrum of an atomic liquid (Xe) is presented. The spectrum is obtained from a simple mode-coupling theory by projecting the dynamics onto bilinear pairs of fluctuating density variables. Good agreement is obtained in comparison with recently reported molecular dynamics simulation results. The microscopic theory provides an understanding of the timescales and molecular motions that govern the two-dimensional signal. Predictions are made for the behavior of the spectrum as a function of temperature and density. The theory shows that novel signatures in the two-dimensional Raman spectrum of supercritical and supercooled liquids are expected. PMID- 11415157 TI - Local-functional theory of critical adsorption. AB - Local-functional methods are applied to critical adsorption in three-dimensional Ising-like systems. The universal order-parameter-profile scaling functions, P(+/ )(x), along the critical isochore (+) and phase boundary (-), are calculated along with their associated universal amplitudes. Good agreement is found with the results of Monte Carlo simulations. General properties of P(+/-)(x) for small and large scaled distance, x, are elucidated. An order-parameter scaling function, P(c)(x), for critical adsorption along the critical isotherm is introduced and calculated. PMID- 11415158 TI - Binary spreading process with parity conservation. AB - Recently there has been a debate concerning the universal properties of the phase transition in the pair contact process with diffusion (PCPD) 2A-->3A, 2A-->0. Although some of the critical exponents seem to coincide with those of the so called parity-conserving universality class, it was suggested that the PCPD might represent an independent class of phase transitions. This point of view is motivated by the argument that the PCPD does not conserve parity of the particle number. In the present work we question what happens if the parity conservation law is restored. To this end, we consider the reaction-diffusion process 2A-->4A, 2A-->0. Surprisingly, this process displays the same type of critical behavior, leading to the conclusion that the most important characteristics of the PCPD is the use of binary reactions for spreading, regardless of whether parity is conserved or not. PMID- 11415159 TI - Periodic-orbit bifurcations and superdeformed shell structure. AB - We have derived a semiclassical trace formula for the level density of the three dimensional spheroidal cavity. To overcome the divergences occurring at bifurcations and in the spherical limit, the trace integrals over the action angle variables were performed using an improved stationary phase method. The resulting semiclassical level density oscillations and shell-correction energies are in good agreement with quantum-mechanical results. We find that the bifurcations of some dominant short periodic orbits lead to an enhancement of the shell structure for "superdeformed" shapes related to those known from atomic nuclei. PMID- 11415160 TI - Synchronization induced by Langevin dynamics. AB - Equilibrium Langevin dynamics of one-dimensional Lennard-Jones chains is studied. It is shown that depending on the noise strength, the friction constant and the number of particles, chains can synchronize, break, or remain desynchronized. Generally the synchronization time and the maximal Lyapunov exponent are found to depend on the number of particles and the ratio of noise strength to friction constant. PMID- 11415161 TI - Multiscale analysis of complex systems. AB - We calculate block information versus size profiles for two-symbol strings generated by several dynamical processes: random, periodic, regular language, and substitutive. The profiles provide a good diagnostic of the complexity of the strings. PMID- 11415162 TI - Subharmonic destruction of generalized chaos synchronization. AB - A bifurcation of transition that destroys generalized chaos synchronization is considered. This transition frequently occurs in regimes of subharmonic chaos entrainment where synchronization can be abruptly terminated due only to an almost unnoticeable change in the shape of the driving attractor. We explore the main cause of this sensitivity and ascertain the mechanism behind this transition. PMID- 11415163 TI - Scaling laws and vortex profiles in two-dimensional decaying turbulence. AB - We use high resolution numerical simulations over several hundred of turnover times to study the influence of small scale dissipation onto vortex statistics in 2D decaying turbulence. A scaling regime is detected when the scaling laws are expressed in units of mean vorticity and integral scale, like predicted in Carnevale et al., Phys. Rev. Lett. 66, 2735 (1991), and it is observed that viscous effects spoil this scaling regime. The exponent controlling the decay of the number of vortices shows some trends toward xi=1, in agreement with a recent theory based on the Kirchhoff model [C. Sire and P. H. Chavanis, Phys. Rev. E 61, 6644 (2000)]. In terms of scaled variables, the vortices have a similar profile with a functional form related to the Fermi-Dirac distribution. PMID- 11415164 TI - Method for measuring local hydraulic permeability using magnetic resonance imaging. AB - The hydraulic permeability of a porous medium characterizes the ease with which a fluid may be driven through it, and is defined via the classical Darcy law. A method for noninvasive, nondestructive measurement of one projection of the local permeability tensor of a porous medium, using gas phase magnetic resonance imaging, is presented. Results are shown for one-dimensional experiments on dry porous rocks. The limitations of the method are explored, and the extension to three-dimensional imaging of the permeability is discussed. PMID- 11415165 TI - Turbulence in a free surface. AB - We report an experimental and numerical study of turbulent fluid motion in a free surface. The flow is realized experimentally on the surface of a tank filled with water stirred by a vertically oscillating grid positioned well below the surface. The effect of surface waves appears to be negligible so that the flow can numerically be realized with a flat surface and stress-free boundary conditions. The surface flow is unconventional in that it is not incompressible and neither energy nor enstrophy are conserved. Nevertheless, according to both experiment and numerical simulation, the second order structure function S2(R) scales essentially as for a three-dimensional system. However, the surface flow seems to be more intermittent. PMID- 11415166 TI - Dynamics of Raman instabilities using chirped laser pulses. AB - Time resolved measurements of the growth of Raman instabilities were performed using a picosecond chirped laser pulse. It was observed experimentally that for a short laser pulse (<10 ps), forward and 30 degrees Raman scattering occur at the back of the pulse. The growth of the instabilities was found to be independent of the sign of the chirp. In addition, a simple temporal model was developed and shows good agreement with the experimental results. This model also indicates that the plasma wave driven by forward Raman scattering is severely damped in the case of pulses longer than a few picoseconds. Damping by the modulational instability is compatible with the experimental results. PMID- 11415167 TI - Area-preserving dynamics of a long slender finger by curvature: a test case for globally conserved phase ordering. AB - A long and slender finger can serve as a simple "test bed" for different phase ordering models. In this work, the globally conserved, interface-controlled dynamics of a long finger is investigated, analytically and numerically, in two dimensions. An important limit is considered when the finger dynamics is reducible to area-preserving motion by curvature. A free boundary problem for the finger shape is formulated. An asymptotic perturbation theory is developed that uses the finger aspect ratio as a small parameter. The leading-order approximation is a modification of the Mullins finger (a well-known analytic solution) whose width is allowed to slowly vary with time. This time dependence is described, in the leading order, by an exponential law with the characteristic time proportional to the (constant) finger area. The subleading terms of the asymptotic theory are also calculated. Finally, the finger dynamics is investigated numerically, employing the Ginzburg-Landau equation with a global conservation law. The theory is in very good agreement with the numerical solution. PMID- 11415168 TI - Pattern formation in a surface chemical reaction with global delayed feedback. AB - We consider effects of global delayed feedback on anharmonic oscillations in the reaction-diffusion model of the CO oxidation reaction on a Pt(110) single-crystal surface. Depending on the feedback intensity and the delay time, we find that various spatiotemporal patterns can be induced. These patterns are characterized using a transformation to phase and amplitude variables designed for anharmonic oscillations. Typical feedback-induced patterns represent traveling phase flips, asynchronous oscillations, and dynamical clustering. Three different types of cluster patterns are identified: amplitude clusters, phase clusters, and cluster turbulence. For phase clusters, two different front instabilities are possible. A pitchfork bifurcation leads to propagation of cluster fronts. An instability of the state of phase balance results in spatial front oscillations. PMID- 11415169 TI - Multichoice minority game. AB - The generalization of the problem of adaptive competition, known as the minority game, to the case of K possible choices for each player, is addressed, and applied to a system of interacting perceptrons with input and output units of a type of K-state Potts spins. An optimal solution of this minority game, as well as the dynamic evolution of the adaptive strategies of the players, are solved analytically for a general K and compared with numerical simulations. PMID- 11415170 TI - Criticality in confined ionic fluids. AB - A theory of a confined two-dimensional electrolyte is presented. The positive and negative ions, interacting by a 1/r potential, are constrained to move on an interface separating two solvents with dielectric constants epsilon(1) and epsilon(2). It is shown that the Debye-Huckel type of theory predicts that this two-dimensional Coulomb fluid should undergo a phase separation into a coexisting liquid (high-density) and gas (low-density) phases. We argue, however, that the formation of polymerlike chains of alternating positive and negative ions can prevent this phase transition from taking place. PMID- 11415171 TI - Homogeneous nucleation of a noncritical phase near a continuous phase transition. AB - Homogeneous nucleation of a new phase near a second, continuous, transition, is considered. The continuous transition is in the metastable region associated with the first-order phase transition, one of whose coexisting phases is nucleating. Mean-field calculations show that as the continuous transition is approached, the size of the nucleus varies as the response function of the order parameter of the continuous transition. This response function diverges at the continuous transition, as does the temperature derivative of the free-energy barrier to nucleation. This rapid drop of the barrier as the continuous transition is approached means that the continuous transition acts to reduce the barrier to nucleation at the first-order transition. This may be useful in the crystallization of globular proteins. PMID- 11415172 TI - Time dependence of breakdown in a global fiber-bundle model with continuous damage. AB - A time-dependent global fiber-bundle model of fracture with continuous damage is formulated in terms of a set of coupled nonlinear differential equations. A first integral of this set is analytically obtained. The time evolution of the system is studied by applying a discrete probabilistic method. Several results are discussed emphasizing their differences with the standard time-dependent model. The results obtained show that with this simple model a variety of experimental observations can be qualitatively reproduced. PMID- 11415173 TI - Density of states, Potts zeros, and Fisher zeros of the Q-state Potts model for continuous Q. AB - The Q-state Potts model can be extended to noninteger and even complex Q by expressing the partition function in the Fortuin-Kasteleyn (F-K) representation. In the F-K representation the partition function Z(Q,a) is a polynomial in Q and v=a-1 (a=e(betaJ)) and the coefficients of this polynomial, Phi(b,c), are the number of graphs on the lattice consisting of b bonds and c connected clusters. We introduce the random-cluster transfer matrix to compute Phi(b,c) exactly on finite square lattices with several types of boundary conditions. Given the F-K representation of the partition function we begin by studying the critical Potts model Z(CP)=Z(Q,a(c)(Q)), where a(c)(Q)=1+square root[Q]. We find a set of zeros in the complex w=square root[Q] plane that map to (or close to) the Beraha numbers for real positive Q. We also identify Q(c)(L), the value of Q for a lattice of width L above which the locus of zeros in the complex p=v/square root[Q] plane lies on the unit circle. By finite-size scaling we find that 1/Q(c)(L)-->0 as L-->infinity. We then study zeros of the antiferromagnetic (AF) Potts model in the complex Q plane and determine Q(c)(a), the largest value of Q for a fixed value of a below which there is AF order. We find excellent agreement with Baxter's conjecture Q(AF)(c)(a)=(1-a)(a+3). We also investigate the locus of zeros of the ferromagnetic Potts model in the complex Q plane and confirm that Q(FM)(c)(a)=(a-1)(2). We show that the edge singularity in the complex Q plane approaches Q(c) as Q(c)(L) approximately Q(c)+AL(-y(q)), and determine the scaling exponent y(q) for several values of Q. Finally, by finite-size scaling of the Fisher zeros near the antiferromagnetic critical point we determine the thermal exponent y(t) as a function of Q in the range 20. AB - We introduce a method of intervals for the analysis of diffusion-limited annihilation, A+A-->0, on the line. The method leads to manageable diffusion equations whose interpretation is intuitively clear. As an example, we treat the following cases: (a) annihilation in the infinite line and in infinite (discrete) chains; (b) annihilation with input of single particles, adjacent particle pairs, and particle pairs separated by a given distance; (c) annihilation, A+A-->0, along with the birth reaction A-->3A, on finite rings, with and without diffusion. PMID- 11415175 TI - Susceptibility and percolation in two-dimensional random field Ising magnets. AB - The ground-state structure of the two-dimensional random field Ising magnet is studied using exact numerical calculations. First we show that the ferromagnetism, which exists for small system sizes, vanishes with a large excitation at a random field strength-dependent length scale. This breakup length scale L(b) scales exponentially with the squared random field, exp(A/delta(2)). By adding an external field H, we then study the susceptibility in the ground state. If L>L(b), domains melt continuously and the magnetization has a smooth behavior, independent of system size, and the susceptibility decays as L-2. We define a random field strength-dependent critical external field value +/ H(c)(delta) for the up and down spins to form a percolation type of spanning cluster. The percolation transition is in the standard short-range correlated percolation universality class. The mass of the spanning cluster increases with decreasing Delta and the critical external field approaches zero for vanishing random field strength, implying the critical field scaling (for Gaussian disorder) H(c) approximately (delta-delta(c))(delta), where delta(c)=1.65+/-0.05 and delta=2.05+/-0.10. Below Delta(c) the systems should percolate even when H=0. This implies that even for H=0 above L(b) the domains can be fractal at low random fields, such that the largest domain spans the system at low random field strength values and its mass has the fractal dimension of standard percolation D(f)=91/48. The structure of the spanning clusters is studied by defining red clusters, in analogy to the "red sites" of ordinary site percolation. The sizes of red clusters define an extra length scale, independent of L. PMID- 11415176 TI - Extremal statistics in the energetics of domain walls. AB - We study at T=0 the minimum energy of a domain wall and its gap to the first excited state, concentrating on two-dimensional random-bond Ising magnets. The average gap scales as deltaE1 approximately L(straight theta)f(N(z)), where f(y) approximately [ln y](-1/2), straight theta is the energy fluctuation exponent, L is the length scale, and N(z) is the number of energy valleys. The logarithmic scaling is due to extremal statistics, which is illustrated by mapping the problem into the Kardar-Parisi-Zhang roughening process. It follows that the susceptibility of domain walls also has a logarithmic dependence on the system size. PMID- 11415177 TI - Time decay of the remanent magnetization in the +/-J spin glass model at T=0. AB - Using the zero-temperature Metropolis dynamics, the time decay of the remanent magnetization in the +/-J Edward-Anderson spin glass model with a uniform random distribution of ferromagnetic and antiferromagnetic interactions has been investigated. Starting from the saturation, the magnetization per spin m reveals a slow decrease with time, which can be approximated by a power law: m(t)=m(infinity)+(t/a(0))(a(1)), a(1)<0. Moreover, its relaxation does not lead it into one of the ground states, and therefore the system is trapped in metastable isoenergetic microstates remaining magnetized. Such behavior is discussed in terms of a random walk that the system performs on its available configuration space. PMID- 11415178 TI - Harris criterion on hierarchical lattices: rigorous inequalities and counterexamples in Ising systems. AB - Random bond Ising systems on a general hierarchical lattice are considered. The inequality between the specific heat exponent of the pure system, alpha(p), and the crossover exponent straight phi, alpha(p)D(B). In this work we perform a finite size scaling analysis of order parameter data at the vicinity of the critical point in dimension d=1. Our results show no signature of a first-order transition in the case of D(A)>D(B). A finite size scaling typical of second-order phase transitions fits well the data from all three regimes. We found that the correlation exponent nu=2 as predicted by field theoretical arguments. Estimates for beta/nu are given for all relevant regimes. PMID- 11415185 TI - Hysteresis and the dynamic phase transition in thin ferromagnetic films. AB - Hysteresis and the nonequilibrium dynamic phase transition in thin magnetic films subject to an oscillatory external field have been studied by Monte Carlo simulation. The model under investigation is a classical Heisenberg spin system with a bilinear exchange anisotropy Lambda in a planar thin film geometry with competing surface fields. The film exhibits a nonequilibrium phase transition between dynamically ordered and dynamically disordered phases characterized by a critical temperature T(cd), whose location is determined by the amplitude H0 and frequency omega of the applied oscillatory field. In the presence of competing surface fields the critical temperature of the ferromagnetic-paramagnetic transition for the film is suppressed from the bulk system value T(c) to the interface localization-delocalization temperature T(ci). The simulations show that in general T(cd)/=2, the rupture dynamics is not defined without the introduction of a regularizing scheme. We investigate three regularization schemes involving, respectively, a saturation of damage, a minimum distance of approach to the crack tip, and a fixed stress maximum. In the first and third schemes, the finite-time singularity is replaced by a crack dynamics defined for all times but which is controlled by either the existence of a microscopic scale at which the stress is regularized or by the maximum sustainable stress. In the second scheme, a finite time singularity is again found. In the first two schemes within this regime m>/=2, the theory has no continuous limit. PMID- 11415196 TI - Monte Carlo simulations of short-time critical dynamics with a conserved quantity. AB - With Monte Carlo simulations, we investigate short-time critical dynamics of the three-dimensional antiferromagnetic Ising model with a globally conserved magnetization m(s) (not the order parameter). From the power law behavior of the staggered magnetization (the order parameter), its second moment and the autocorrelation, we determine all static and dynamic critical exponents as well as the critical temperature. The universality class of m(s)=0 is the same as that without a conserved quantity, but the universality class of nonzero m(s) is different. PMID- 11415197 TI - Rotation-induced phase transition in a spherical gravitating system. AB - Due to the infinite range and singularity of the gravitational force, it is difficult to directly apply the standard methods of statistical physics to self gravitating systems, e.g., interstellar grains, globular clusters, galaxies, etc. Unusual phenomena can occur, such as a negative heat capacity, unbounded mass, or the gravothermal catastrophe where the equilibrium state is fully collapsed and the entropy is unbounded. Using mean field theory, we investigate the influence of rotation on a purely spherical gravitational system. Although spherical symmetry nullifies the total angular momentum, its square is finite and conserved. Here we study the case where each particle has specific angular momentum of the same magnitude l. We rigorously prove the existence of an upper bound on the entropy and a lower bound for the energy. We demonstrate that, in the microcanonical and canonical ensembles, a phase transition occurs when l falls below a critical value. We characterize the properties of each phase and construct the coexistence curve for each ensemble. Possible applications to astrophysics are considered. PMID- 11415198 TI - Competitive growth model involving random deposition and random deposition with surface relaxation. AB - A deposition model that considers a mixture of random deposition with surface relaxation and a pure random deposition is proposed and studied. As the system evolves, random deposition with surface relaxation (pure random deposition) take place with probability p and (1-p), respectively. The discrete (microscopic) approach to the model is studied by means of extensive numerical simulations, while continuous equations are used in order to investigate the mesoscopic properties of the model. A dynamic scaling ansatz for the interface width W(L,t,p) as a function of the lattice side L, the time t and p is formulated and tested. Three exponents, which can be linked to the standard growth exponent of random deposition with surface relaxation by means of a scaling relation, are identified. In the continuous limit, the model can be well described by means of a phenomenological stochastic growth equation with a p-dependent effective surface tension. PMID- 11415199 TI - Roughness exponent in two-dimensional percolation, Potts model, and clock model. AB - We present a numerical study of the self-affine profiles obtained from configurations of the q-state Potts (with q=2,3, and 7) and p=10 clock models as well as from the occupation states for site percolation on the square lattice. The first and second order static phase transitions of the Potts model are located by a sharp change in the value of the roughness exponent alpha characterizing those profiles. The low temperature phase of the Potts model corresponds to flat (alpha approximately 1) profiles, whereas its high temperature phase is associated with rough (alpha approximately 0.5) ones. For the p=10 clock model, in addition to the flat (ferromagnetic) and rough (paramagnetic) profiles, an intermediate rough (0.5 or = l(y). Here, l(parallel) (l(perpendicular)) is the correlation length along (perpendicular to) the average field B(0)=B(0)e(z). We have anomalous, non-Gaussian transport for R less, similar 0.1, in which case the mean square deviation scales nonlinearly with time. For R greater, similar 1 we have several Gaussian regimes: an almost quasilinear regime for 0.1 less, similar R less, similar 1, an intermediate, transition regime for 1 less, similar R less, similar 10, and a percolative regime for R greater, similar 10. An analytical form of the diffusion coefficient is proposed, D(i)=D(deltaBl(z)/B(0)l(x))(mu)(l(i)/l(x))(nu)l(2)(x)/l(z), which well describes the numerical simulation results in the quasilinear, intermediate, and percolative regimes. PMID- 11415234 TI - Kinetic approach to low-frequency waves in dusty self-gravitating plasmas. AB - A kinetic model is derived for the propagation of low-frequency waves in a dusty plasma containing very heavy dust particles, when the self-gravitational interaction due to these grains is included in the analysis. Analytical expressions for the dispersion function are used to examine the instability and damping of the modes. The stability regions of low-frequency waves are compared in the kinetic and the analogous hydrodynamic models, showing that there are only slight differences. However, the kinetic analysis modifies the growth rates of the Jeans instability and can considerably alter the conditions for the propagation of stable dust modes. PMID- 11415235 TI - Traveling-wave tubes and backward-wave oscillators with weak external magnetic fields. AB - Recent development of plasma-assisted slow-wave oscillators [Goebel et al. IEEE Trans. Plasma Sci. 22, 547 (1994)], microwave sources that operate without guiding magnetic fields, has stimulated interest in the theoretical analysis of such tubes. In principle, in the absence of guiding magnetic fields, due to the space charge forces and the radial electric field of the wave, the electrons may propagate radially outward which increases electron coupling to the slow wave whose field is localized near the slow-wave structure (SWS). This increases the wave growth rate and efficiency, and hence allows one to shorten the interaction region. So the radial electron motion can be beneficial for operation if it does not lead to interception of electrons by the SWS. To avoid this interception a weak external magnetic field can be applied. The theory developed describes the effect of weak magnetic fields on the operation of traveling-wave tubes and backward-wave oscillators with electrons moving not only axially but also transversely. This theory allows one to estimate the magnetic field required for protecting the SWS from electron bombardment at different power levels. Theoretical predictions of the efficiency enhancement due to the weak magnetic field are confirmed in experiments. PMID- 11415236 TI - Polarization state of quadratic spatial optical solitons. AB - We derive governing equations that determine a full polarization state of transversely two-dimensional spatial solitons in a bulk anisotropic medium with the second-order nonlinearity. Based on nonlinear vectorial Maxwell's equations and approximation of slowly varying envelopes, our approach describes also lowest order nonparaxial effects, however the most important factor governing radiation polarization is the medium anisotropy. This factor results in mixing of orthogonal components of electric field of quadratic soliton that consists of coupled beams at the fundamental frequency and its second harmonics. For the case of weak anisotropy we determine the soliton polarization state by a perturbation method; it turns out that it is elliptical and changing over the soliton transverse section. The approach allows generalization to the case of optical parametric oscillators. PMID- 11415237 TI - Universal criterion and amplitude equation for a nonequilibrium Ising-Bloch transition. AB - We identify a universal criterion for the onset of a nonequilibrium Ising-Bloch (NIB) transition, and describe the behavior near the bifurcation by a generic amplitude equation. We found that a NIB transition is caused by an antisymmetric eigenvector passing the translational mode of the system at a critical point. In this context we discuss Hamiltonian and dissipative systems. We report on a NIB in nonlinear optics, manifesting itself in a transition from static to moving polarization fronts. PMID- 11415238 TI - Discrete breathers in dissipative lattices. AB - We study the properties of discrete breathers, also known as intrinsic localized modes, in the one-dimensional Frenkel-Kontorova lattice of oscillators subject to damping and external force. The system is studied in the whole range of values of the coupling parameter, from C=0 (uncoupled limit) up to values close to the continuum limit (forced and damped sine-Gordon model). As this parameter is varied, the existence of different bifurcations is investigated numerically. Using Floquet spectral analysis, we give a complete characterization of the most relevant bifurcations, and we find (spatial) symmetry-breaking bifurcations that are linked to breather mobility, just as it was found in Hamiltonian systems by other authors. In this way moving breathers are shown to exist even at remarkably high levels of discreteness. We study mobile breathers and characterize them in terms of the phonon radiation they emit, which explains successfully the way in which they interact. For instance, it is possible to form "bound states" of moving breathers, through the interaction of their phonon tails. Over all, both stationary and moving breathers are found to be generic localized states over large values of C, and they are shown to be robust against low temperature fluctuations. PMID- 11415239 TI - Stability of stationary states in the cubic nonlinear Schrodinger equation: applications to the Bose-Einstein condensate. AB - The cubic nonlinear Schrodinger equation is the quasi-one-dimensional limit of the mean-field theory which models dilute gas Bose-Einstein condensates. Stationary solutions of this equation can be characterized as soliton trains. It is demonstrated that for repulsive nonlinearity a soliton train is stable to initial stochastic perturbation, while for attractive nonlinearity its behavior depends on the spacing between individual solitons in the train. Toroidal and harmonic confinement, both of experimental interest for Bose-Einstein condensates, are considered. PMID- 11415240 TI - Stopping of acoustic waves by sonic polymer-fluid composites. AB - A two-dimensional periodic array of air cylinders in water is known to have giant acoustic stop bands [M.S. Kushwaha and B. Djafari-Rouhani, J. Appl. Phys. 84 (1998) 4677]. It is shown in the present paper that hollow cylinders made of an elastically-soft polymer containing air inside and arranged on a square lattice in water can still give rise to large acoustic band gaps. Similar properties can also be obtained with a close-packed array of tubes containing water when arranged on a honeycomb lattice in air. The transmission coefficient of films made of such polymer-fluid composites has been calculated by finite difference time domain method. With film thickness not exceeding 75 mm, a deep sonic attenuation band was found with, in the best cases, a lower limit below 1 kHz and an upper limit above 10 kHz. PMID- 11415241 TI - Quantization of weakly nonlinear lattices: envelope solitons. AB - A method of quantizing weakly nonlinear lattices is proposed. It is based on introducing "pseudofield" operators. In this formalism quantum envelope solitons together with phonons are regarded as elementary quasiparticles making up a boson gas. In the classical limit the excitations corresponding to frequencies above a linear cutoff frequency are reduced to conventional envelope solitons. The approach allows one to identify a quantum soliton that is localized in space and to understand the existence of a narrow soliton frequency band. PMID- 11415242 TI - Nonadiabatic interaction of a propagating wave packet with localized parametric pumping. AB - The interaction of a propagating wave packet (carrier frequency omega, wave number k) with nonadiabatic parametric pumping, localized in a region of size the order of the carrier wavelength L approximately lambda=2pi/k, is studied experimentally in a system of dipolar spin waves in a ferrite film. It is shown that the three-wave parametric interaction omega+omega(')=omega(p) leads to the formation of both contrapropagating (k(')=-k) and copropagating (k(')=k) idle wave packets of carrier frequency omega('). A system of equations derived for the packet envelopes gives a quantitative description of the observed random modulation of the output signal caused by the interference of the input and copropagating idle wave packets. PMID- 11415243 TI - Interactions between two-dimensional composite vector solitons carrying topological charges. AB - We present a comprehensive study of interactions (collisions) between two dimensional composite vector solitons carrying topological charges in isotropic saturable nonlinear media. We numerically study interactions between such composite solitons for different regimes of collision angle and report numerous effects which are caused solely by the "spin" (topological charge) carried by the second excited mode. The most intriguing phenomenon we find is the delayed-action interaction between interacting composite solitons carrying opposite spins. In this case, two colliding solitons undergo a fusion process and form a metastable bound state that decays after long propagation distances into two or three new solitons. Another noticeable effect is spin-orbit coupling in which angular momentum is being transferred from "spin" to orbital angular momentum. This phenomenon occurs at angles below the critical angle, including the case when the initial soliton trajectories are in parallel to one another and lie in the same plane. Finally, we report on shape transformation of vortex component into a rotating dipole-mode solitons that occurs at large collision angles, i.e., at angles for which scalar solitons of all types simply go through one another unaffected. PMID- 11415244 TI - Nonreciprocal magnetic photonic crystals. AB - We study band dispersion relations omega(k-->) of a photonic crystal with at least one of the constitutive components being a magnetically ordered material. It is shown that by proper spatial arrangement of magnetic and dielectric components one can construct a magnetic photonic crystal with strong spectral asymmetry (nonreciprocity) omega(k-->) not equal omega(-k-->). The spectral asymmetry, in turn, results in a number of interesting phenomena, in particular, one-way transparency when the magnetic photonic crystal, being perfectly transparent for a Bloch wave of frequency omega, "freezes" the radiation of the same frequency omega propagating in the opposite direction. The frozen radiation corresponds to a Bloch wave with zero group velocity partial differential omega(k)/ partial differential k=0 and, in addition, with partial differential(2)omega(k)/ partial differential k(2)=0. PMID- 11415245 TI - Walking cavity solitons. AB - A family of walking solitons is obtained for the degenerate optical parametric oscillator below threshold. The loss-driven mechanism of velocity selection for these structures is described analytically and numerically. Our approach is based on understanding the role played by the field momentum and generic symmetry properties and, therefore, it can be easily generalized to other dissipative multicomponent models with walk off. PMID- 11415246 TI - Acoustic wave propagation in a one-dimensional layered system. AB - Propagation of acoustic waves in a one-dimensional water duct containing many air filled blocks is studied by the transfer matrix formalism. Energy distribution and interface vibration of the air blocks are computed. For periodic arrangement, band structure is calculated analytically, whereas the Lyapunov exponent and its variance are computed numerically for random situations. A distinct collective behavior for localized waves is found. The results are also compared with optical situations. PMID- 11415247 TI - Energy eigenstates of magnetostatic waves and oscillations. AB - Effect of excitation of magnetostatic oscillations in a ferrite resonator by the microwave magnetic field was a subject of many publications of more than the last 40 years. The most interesting multiresonance spectrum of absorption peaks one can observe experimentally is a case of disk-form small ferrite resonators. It is shown in this paper that such small ferrite resonators can be considered as "artificial molecular structures" with properties characterized by energy eigenstates of magnetostatic oscillations. A special interest in these properties may be found in the field of microwave artificial composite materials. PMID- 11415248 TI - Dynamics of kinks in one- and two-dimensional hyperbolic models with quasidiscrete nonlinearities. AB - We study the evolution of fronts in the Klein-Gordon equation when the nonlinear term is inhomogeneous. Extending previous works on homogeneous nonlinear terms, we describe the derivation of an equation governing the front motion, which is strongly nonlinear, and, for the two-dimensional case, generalizes the damped Born-Infeld equation. We study the motion of one- and two-dimensional fronts finding a much richer dynamics than in the homogeneous system case, leading, in most cases, to the stabilization of one phase inside the other. For a one dimensional front, the function describing the inhomogeneity of the nonlinear term acts as a "potential function" for the motion of the front, i.e., a front initially placed between two of its local maxima asymptotically approaches the intervening minimum. Two-dimensional fronts, with radial symmetry and without dissipation can either shrink to a point in finite time, grow unboundedly, or their radius can oscillate, depending on the initial conditions. When dissipation effects are present, the oscillations either decay spirally or not depending on the value of the damping dissipation parameter. For fronts with a more general shape, we present numerical simulations showing the same behavior. PMID- 11415249 TI - One-dimensional toy model of globular clusters. AB - We introduce a one-dimensional toy model of globular clusters. The model is a version of the well-known gravitational sheets system, where we also take into account mass and energy loss by evaporation of stars at the boundaries. Numerical integration by the "exact" event-driven dynamics is performed, for initial uniform density and Gaussian random velocities. Two distinct quasistationary asymptotic regimes are attained, depending on the initial energy of the system. We guess the forms of the density and velocity profiles that fit numerical data extremely well and allow us to perform an independent calculation of the self consistent gravitational potential. Some power laws for the asymptotic number of stars and for the collision times are suggested. PMID- 11415250 TI - Green's tensor technique for scattering in two-dimensional stratified media. AB - We present an accurate and self-consistent technique for computing the electromagnetic field in scattering structures formed by bodies embedded in a stratified background and extending infinitely in one direction (two-dimensional geometry). With this fully vectorial approach based on the Green's tensor associated with the background, only the embedded scatterers must be discretized, the entire stratified background being accounted for by the Green's tensor. We first derive the formulas for the computation of this dyadic and discuss in detail its physical substance. The utilization of this technique for the solution of scattering problems in complex structures is then illustrated with examples from photonic integrated circuits (waveguide grating couplers with varying periodicity). PMID- 11415251 TI - Role of polarization mode dispersion on modulational instability in optical fibers. AB - We introduce the theory of modulational instability (MI) of electromagnetic waves in fibers with random polarization mode dispersion. Applying a linear stability analysis and stochastic calculus, we show that the MI gain spectrum reads as the maximal eigenvalue of a constant effective matrix. In the limiting cases of small or large fluctuations, we give explicit expressions for the MI gain spectra. In the general configurations, we give the explicit form of the effective matrix and numerically compute the maximal eigenvalue. In the anomalous dispersion regime, polarization dispersion widens the unstable bandwidth. Depending on the type of variations of the birefringence parameters, polarization dispersion reduces or enhances the MI gain peak. In the normal dispersion regime, random effects may extend the instability domain to the whole spectrum of modulations. The linear stability analysis is confirmed by numerical simulation of the full stochastic coupled nonlinear Schrodinger equations. PMID- 11415252 TI - Efficient reconstruction of multiphase morphologies from correlation functions. AB - A highly efficient algorithm for the reconstruction of microstructures of heterogeneous media from spatial correlation functions is presented. Since many experimental techniques yield two-point correlation functions, the restoration of heterogeneous structures, such as composites, porous materials, microemulsions, ceramics, or polymer blends, is an inverse problem of fundamental importance. Similar to previously proposed algorithms, the new method relies on Monte Carlo optimization, representing the microstructure on a discrete grid. An efficient way to update the correlation functions after local changes to the structure is introduced. In addition, the rate of convergence is substantially enhanced by selective Monte Carlo moves at interfaces. Speedups over prior methods of more than two orders of magnitude are thus achieved. Moreover, an improved minimization protocol leads to additional gains. The algorithm is ideally suited for implementation on parallel computers. The increase in efficiency brings new classes of problems within the realm of the tractable, notably those involving several different structural length scales and/or components. PMID- 11415253 TI - Thirteen-velocity three-dimensional lattice Boltzmann model. AB - A thirteen-velocity three-dimensional lattice Boltzmann model on a cubic grid is presented. The transport coefficients derived from the standard Chapman-Enskog expansion are given together with the conditions for isotropy and Galilean invariance. The different invariants of the model are discussed. The results of measurements of drag and torque on a free falling sphere in a cylinder are in good agreement with solutions of the Navier-Stokes equation. Comparison of the time evolution of a freely decaying Taylor-Green vortex computed by fast Fourier transform and by the present model is presented. PMID- 11415254 TI - Nystrom plus correction method for solving bound-state equations in momentum space. AB - A method is presented for solving the momentum-space Schrodinger equation with a linear potential. The Lande-subtracted momentum-space integral equation can be transformed into a matrix equation by the Nystrom method. The method produces only approximate eigenvalues in the cases of singular potentials such as the linear potential. The eigenvalues generated by the Nystrom method can be improved by calculating the numerical errors and adding the appropriate corrections. The end results are more accurate eigenvalues than those generated by the basis function method. The method is also shown to work for a relativistic equation such as the Thompson equation. PMID- 11415255 TI - Directed self-avoiding walks in random media. AB - Two types of directed self-avoiding walks (SAW's), namely, three-choice directed SAW and outwardly directed SAW, have been studied on infinite percolation clusters on the square lattice in two dimensions. The walks on the percolation clusters are generated via a Monte Carlo technique. The longitudinal extension R(N) and the transverse fluctuation W(N) have been measured as a function of the number of steps N. Slight swelling is observed in the longitudinal direction on the random lattices. A crossover from shrinking to swelling of the transverse fluctuations is found at a certain length N(c) of the walks. The exponents related to the transverse fluctuations are seen to be unchanged in the random media even as the percolation threshold is reached. The scaling function form of the extensions are verified. PMID- 11415256 TI - Anomalous relaxation and self-organization in nonequilibrium processes. AB - We study thermal relaxation in ordered arrays of coupled nonlinear elements with external driving. We find that our model exhibits dynamic self-organization manifested in a universal stretched-exponential form of relaxation. We identify two types of self-organization, cooperative and anticooperative, which lead to fast and slow relaxation, respectively. We give a qualitative explanation for the behavior of the stretched exponent in different parameter ranges. We emphasize that this is a system exhibiting stretched-exponential relaxation without explicit disorder or frustration. PMID- 11415257 TI - Extremal principle for the steady-state selection in driven lattice gases with open boundaries. AB - This paper investigates the steady states of one-dimensional driven lattice gases with open boundary conditions. It shows how the extremal principle proposed recently by Popkov and Schutz can be modified to apply to more general cases. Monte Carlo simulations are presented for a one-dimensional totally asymmetric simple exclusion process with nearest neighbor repulsion under parallel update as an example. The simulations enable one to guess the exact phase diagram for this particular lattice gas with deterministic bulk dynamics, by fitting the data to analytic formulas, which appear to be exact in the thermodynamic limit. PMID- 11415258 TI - Phase transition of the one-dimensional coagulation-production process. AB - Recently an exact solution has been found by M. Henkel and H. Hinrichsen [J. Phys. A 34, 1561 (2001)] for the one-dimensional coagulation-production process: 2A-->A, AOA-->3A with equal diffusion and coagulation rates. This model evolves into the inactive phase independently of the production rate with t(-1/2) density decay law. This paper shows that cluster mean-field approximations and Monte Carlo simulations predict a continuous phase transition for higher diffusion/coagulation rates as considered by the exact solution. Numerical evidence is given that the phase transition universality agrees with that of the annihilation-fission model with low diffusions. PMID- 11415259 TI - Controlling spatiotemporal chaos in coupled map lattices. AB - A simple method is presented for controlling spatiotemporal chaos in coupled map lattices to a homogeneous state. This method can be applied to many kinds of models such as coupled map lattices (CML), one-way open CML (the open-flow model), and globally coupled map. We offer the stability analysis of the homogeneous state. Simple and sufficient conditions are obtained for controlling the above mentioned models. Our theoretical results agree well with numerical simulations. PMID- 11415260 TI - Mechanical model for the plasma maser effect. AB - A Fermi-like pinball model is proposed for the nonlinear plasma maser effect, or turbulent bremsstrahlung, in the nonlinear interaction of plasma particles and waves. The model consists of a system of many noninteracting particles bouncing elastically between two oscillating walls. The walls act as energy and momentum sources and sinks for the particles, analogous to the wave fields in a weakly turbulent plasma. The oscillation amplitudes and frequencies of the walls determine the dynamics and distribution of the particles. The resulting asymptotic velocity distributions agree qualitatively with existing weak turbulence theories. It is also found that the second wall, which simulates the effect of the nonresonant wave-particle interaction, can destroy correlations in the particle dynamics and lead to the formation of a high-energy tail in the velocity distribution. PMID- 11415261 TI - Dislocation-mediated melting: the one-component plasma limit. AB - The melting parameter Gamma(m) of a classical one-component plasma is estimated using a relation between the melting temperature, density, shear modulus, and a crystal coordination number that follows from our model of dislocation-mediated melting. We obtain gamma(m)=172+/-35, in good agreement with the results of numerous Monte Carlo calculations. PMID- 11415262 TI - Comment on "Models of intermediate spectral statistics". AB - In this Comment we point out that the semi-Poisson is well suited only as a reference point for the so-called "intermediate statistics," which cannot be interpreted as a universal ensemble, like the Gaussian orthogonal ensemble or the Poissonian statistics. In Ref. 2 it was proposed that the nearest-neighbor distribution P(s) of the spectrum of a Poissonian distributed matrix perturbed by a rank one matrix is similar to the semi-Poisson distribution. We show, however, that the P(s) of this model differs considerably in many aspects from the semi Poisson. In addition, we give an asymptotic formula for P(s) as s-->0, which gives P'(0)=pisqrt[3]/2 for the slope at s=0. This is different not only from the GOE case, but also from the semi-Poisson prediction. PMID- 11415263 TI - Pauli blocking of collisions in a quantum degenerate atomic Fermi gas. AB - We have produced an interacting quantum degenerate Fermi gas of atoms composed of two spin states of magnetically trapped 40K. The relative Fermi energies are adjusted by controlling the population in each spin state. Thermodynamic measurements reveal a resulting imbalance in the mean energy per particle between the two species, which is a factor of 1.4 at our lowest temperature. This imbalance of energy comes from a suppression of collisions between atoms in the gas due to the Pauli exclusion principle. Through measurements of the thermal relaxation rate we have directly observed this Pauli blocking as a factor of 2 reduction in the effective collision cross section in the quantum degenerate regime. PMID- 11415264 TI - Bosons in cigar-shaped traps: Thomas-Fermi regime, Tonks-Girardeau regime, and in between. AB - We present a quantitative analysis of the experimental accessibility of the Tonks Girardeau gas in present-day experiments with cigar-trapped alkalis. For this purpose we derive, using a Bethe ansatz generated local equation of state, a set of hydrostatic equations describing one-dimensional, delta-interacting Bose gases trapped in a harmonic potential. The resulting solutions cover the entire range of atomic densities. PMID- 11415265 TI - Coupled normal heat and matter transport in a simple model system. AB - We introduce the first simple mechanical system that shows fully realistic transport behavior while still being exactly solvable at the level of equilibrium statistical mechanics. The system is a Lorentz gas with fixed freely rotating circular scatterers which scatter point particles via perfectly rough collisions. Upon imposing either a temperature gradient and/or a chemical potential gradient, a stationary state is attained for which local thermal equilibrium holds. Transport in this system is normal in the sense that the transport coefficients which characterize the flow of heat and matter are finite in the thermodynamic limit. Moreover, the two flows are nontrivially coupled, satisfying Onsager's reciprocity relations. PMID- 11415266 TI - Magnetic tension and the geometry of the universe. AB - The vector nature of magnetic fields and the geometrical interpretation of gravity introduced by general relativity guarantee a special coupling between magnetism and spacetime curvature. This magnetogeometrical interaction effectively transfers the tension properties of the field into the spacetime fabric, triggering a variety of effects with profound implications. Given the ubiquity of magnetic fields in the universe, these effects could prove critical. We discuss the nature of the magnetic-field--curvature coupling and illustrate some of its potential implications for cosmology. PMID- 11415267 TI - Measurements of the rare decay K(L) --> e(+)e(-)e(+)e(-). AB - We observe 441 K(L)-->e(+)e(-)e(+)e(-) candidate events with a background of 4.2 events and measure B(K(L)-->e(+)e(-)e(+)e(-)) = [3.72+/-0.18(stat)+/ 0.23(syst)]x10(-8) in the KTeV/E799II experiment at Fermilab. Using the distribution of the angle between the planes of the e(+)e(-) pairs, we measure the CP parameters beta(CP) = -0.23+/-0.09(stat)+/-0.02(syst) and gamma(CP) = 0.09+/-0.09(stat)+/-0.02(syst). We also present the first detailed study of the e(+)e(-) invariant mass spectrum in this decay mode. PMID- 11415268 TI - First measurement of the low- x, low- Q(2) structure function F(2) in neutrino scattering. AB - A new structure function analysis of CCFR deep inelastic nu-N and nu-N scattering data is presented for previously unexplored kinematic regions down to Bjorken x = 0.0045 and Q(2) = 0.3 GeV(2). Comparisons to charged lepton scattering data from NMC and E665 experiments are made and the behavior of the structure function F(2)(nu)2 is studied in the limit Q(2)-->0. PMID- 11415269 TI - Does the D(-)/D(+) production asymmetry decrease at large x(F)? AB - We have applied the meson cloud model to calculate the asymmetries in D and D(s) meson production in high-energy Sigma(-)-nucleus and pi(-)-nucleus collisions. We find a good agreement with recent data. Our results suggest that the asymmetries may decrease at large x(F). PMID- 11415270 TI - Kinetic equation with exact charge conservation. AB - A kinetic master equation for multiplicity distributions is formulated for charged particles which are created or destroyed only in pairs due to the conservation of their Abelian charge. It allows one to study time evolution of the multiplicity distributions in a relativistic many-body system with arbitrary average particle multiplicities. It is shown to reproduce the equilibrium results for both canonical (rare particles) and grand canonical (abundant particles) systems. For canonical systems, the equilibrium multiplicity is much lower and the relaxation time is much shorter than the naive extrapolation from grand canonical results. Implications for chemical equilibration in heavy-ion collisions are also discussed. PMID- 11415271 TI - Photoneutron cross sections for unstable neutron-rich oxygen isotopes. AB - The dipole response of stable and unstable neutron-rich oxygen nuclei of masses A = 17 to A = 22 has been investigated experimentally utilizing electromagnetic excitation in heavy-ion collisions at beam energies about 600 MeV/nucleon. A kinematically complete measurement of the neutron decay channel in inelastic scattering of the secondary beam projectiles from a Pb target was performed. Differential electromagnetic excitation cross sections d sigma/dE were derived up to 30 MeV excitation energy. In contrast to stable nuclei, the deduced dipole strength distribution appears to be strongly fragmented and systematically exhibits a considerable fraction of low-lying strength. PMID- 11415272 TI - Measurement of the elastic magnetic form factor of (3)He at high momentum transfer. AB - New electron scattering measurements have been made that extend data on the (3)He elastic magnetic form factor up to Q(2) = 42.6 fm(-2). These new data test theoretical conjectures regarding non-nucleonic effects in the three-body system. The very small cross sections, as low as 10(-40) cm(2)/sr, required the use of a high-pressure cryogenic gas target and a detector system with excellent background rejection capability. No existing theoretical calculation satisfactorily accounts for all the available data. PMID- 11415273 TI - Stability of negatively charged ions moving in a magnetic field. AB - In a magnetic field the center of mass (c.m.) motion of an atom or molecule couples to the electronic motion. It is demonstrated that this coupling dramatically influences the properties of negative ions. Neglecting c.m. effects the external field gives rise to a series of infinitely many bound states of the ion. Center of mass effects terminate this series and turn bound states into short-lived resonance states. Whether bound states exist at all, their number and properties as well as the lifetimes of the resonance states depend on the neutral system to which an electron is attached, and on the magnetic field. PMID- 11415274 TI - Ab initio investigation of the phase lag in coherent control of H(2). AB - An ab initio investigation of the excitation of an H(2) resonance, coupled to ionization and dissociation continua, was performed. The excitation occurs via a one- and a three-photon path where both fields have a variable but well-defined phase relation. While both continuum yields show an oscillatory behavior as the phase is varied, a pronounced phase lag between the two oscillations is observed, if the excitation energy is detuned from the resonant transition. The results are discussed in connection to a recent series of experiments performed on HI and the subsequent discussion on the possible origin of the observed phase lag. PMID- 11415275 TI - Attosecond time-scale intra-atomic phase matching of high harmonic generation. AB - Using a model of high-harmonic generation that couples a fully quantum calculation with a semiclassical electron trajectory picture, we show that a new type of phase matching is possible when an atom is driven by an optimal optical waveform. For an optimized laser pulse shape, strong constructive interference is obtained in the frequency domain between emissions from different electron trajectories, thereby selectively enhancing a particular harmonic order. This work demonstrates that coherent control in the strong-field regime is possible by adjusting the peaks of a laser field on an attosecond time scale. PMID- 11415276 TI - Nonlinear optics with phase-controlled pulses in the sub-two-cycle regime. AB - Nonlinear optical effects due to the phase between carrier and envelope are observed with 5 fs pulses from a Kerr-lens mode-locked Ti:sapphire laser. These sub-two-cycle pulses with octave spanning spectra are the shortest pulses ever generated directly from a laser oscillator. Detection of the carrier-envelope phase slip is made possible by simply focusing the short pulses directly from the oscillator into a BBO crystal. As a further example of nonlinear optics with such short pulses, the interference between second- and third-harmonic components is also demonstrated. PMID- 11415277 TI - Electromagnetic wave chaos in gradient refractive index optical cavities. AB - Electromagnetic wave chaos is investigated using two-dimensional optical cavities formed in a cylindrical gradient refractive index lens with reflective surfaces. When the planar ends of the lens are cut at an angle to its axis, the geometrical ray paths are chaotic. In this regime, the electromagnetic mode spectrum of the cavity is modulated by both real and ghost periodic ray paths, which also "scar" the electric field intensity distributions of many modes. When the cavity is coupled to waveguides, the eigenmodes generate complex series of resonant peaks in the electromagnetic transmission spectrum. PMID- 11415278 TI - Breakup and fusion of self-guided femtosecond light pulses in air. AB - We report experiments showing the breakup and the merging of filaments formed by the modulational instability of femtosecond optical pulses in air. For input powers as high as 25 times the self-focusing threshold, the beams are shown to split into two spots, which coalesce into a self-guided beam. This effect occurs in an optically Kerr regime and plays an important role in the guiding process. Numerical simulations and theoretical estimates both support the comparison with the experimental data. PMID- 11415279 TI - Self-trapping and stable localized modes in nonlinear photonic crystals. AB - We predict the existence of stable nonlinear localized modes near the band edge of a two-dimensional reduced-symmetry photonic crystal with a Kerr nonlinearity. Employing the technique based on the Green function, we reveal a physical mechanism of the mode stabilization associated with the effective nonlinear dispersion and long-range interaction in the photonic crystals. PMID- 11415280 TI - Impenetrable barriers in phase-space. AB - Dynamical systems theory is used to construct a general phase-space version of transition state theory. Special multidimensional separatrices are found which act as impenetrable barriers in phase-space between reacting and nonreacting trajectories. The elusive momentum-dependent transition state between reactants and products is thereby characterized. A practical algorithm is presented and applied to a strongly coupled Hamiltonian. PMID- 11415281 TI - Breakdown of universality in transitions to spatiotemporal chaos. AB - We show that the transition from laminar to active behavior in extended chaotic systems can vary from a continuous transition in the universality class of directed percolation with infinitely many absorbing states to what appears as a first-order transition. The latter occurs when finite lifetime nonchaotic structures, called "solitons," dominate the dynamics. We illustrate this scenario in an extension of the deterministic Chate-Manneville coupled map lattice model and in a soliton including variant of the stochastic Domany-Kinzel cellular automaton. PMID- 11415282 TI - Simple one-dimensional model of heat conduction which obeys Fourier's law. AB - We present the computer simulation results of a chain of hard-point particles with alternating masses interacting on its extremes with two thermal baths at different temperatures. We found that the system obeys Fourier's law at the thermodynamic limit. This result is against the actual belief that one dimensional systems with momentum conservative dynamics and nonzero pressure have infinite thermal conductivity. It seems that thermal resistivity occurs in our system due to a cooperative behavior in which light particles tend to absorb much more energy than the heavier ones. PMID- 11415283 TI - Avalanches in one-dimensional piles with different types of bases. AB - We perform a systematic experimental study of the influence of the type of base on the avalanche dynamics of slowly driven 1D ball piles. The control of base details allows us to explore a wide spectrum of pile structures and dynamics. The scaling properties of the observed avalanche distributions suggest that self organized critical behavior is approached as the "base-induced" disorder at the pile profile increases. PMID- 11415284 TI - Translation drag coefficient of a self-similar assembly of spheres immersed in an incompressible fluid. AB - On the basis of deterministic fractals and the Rotne-Prager hydrodynamic interaction tensor, we confirm the asymptotic as well as the finite size scaling of the friction coefficient lambda of a self-similar structure. The fractal assembly is made of N spheres with its dimension varying from D < 1 to D = 3. The number of spheres can be as high as N approximately O(10(4)). The asymptotic scaling behavior of the friction coefficient per sphere is lambda approximately N(1/D-1) for D > 1, lambda approximately (lnN)(-1) for D = 1, and lambda approximately N(0) for D < 1. The crossover behavior indicates that while in the regime of D > 1 the hydrodynamic screening effect grows with the size, for D<1 it is limited in a finite range, which decays with decreasing D. PMID- 11415285 TI - Experimental characterization of the electron heat transport in low-density ASDEX upgrade plasmas. AB - The electron heat transport is investigated in ASDEX Upgrade conventional L-mode plasmas with pure electron heating provided by electron-cyclotron heating (ECH) at low density. Under these conditions, steady-state and ECH modulation experiments indicate without ambiguity that electron heat transport exhibits a clear threshold in inverted Delta T(e)/T(e) and also suggest that it has a gyro Bohm character. PMID- 11415286 TI - Nonlocal power deposition in inductively coupled plasmas. AB - Radiofrequency (rf) plasmas exhibit field penetration well beyond the classical skin depth. Two physical explanations are proposed. First, by tracing orbits of electrons through many rf cycles in a cylindrical system, it is shown that numerous ionizing electrons can reach the interior. Second, current-carrying electrons can form a long-lived torus that drifts toward the axis, causing frequently observed interference phenomena. The pressure dependence of this effect does not agree with collisionless theories of anomalous skin effect, but is consistent with the proposed mechanism. PMID- 11415287 TI - Acoustic crystallization and heterogeneous nucleation. AB - By focusing a high-intensity acoustic wave in liquid helium, we have observed the nucleation of solid helium inside the wave above a certain threshold in amplitude. The nucleation is a stochastic phenomenon. Its probability increases continuously from 0 to 1 in a narrow pressure interval around P(m) + 4.7 bars ( P(m) = 25.3 bars is the melting pressure where liquid and solid helium are in equilibrium). This overpressure is larger by 2 to 3 orders of magnitude than what had been previously observed. Our result strongly supports the recent suggestion by Balibar, Mizusaki, and Sasaki that, in all previous experiments, solid helium nucleated on impurities. PMID- 11415288 TI - Atomic string holography. AB - A new diffraction-channeling effect has been discovered, in which Kikuchi or channeling line patterns formed by high energy electrons, neutrons, and positrons are shown to break up into a series of annular disks if the crystal thickness traversed by the beam is small. The disks may be interpreted as Gabor in-line holograms of strings of atoms projected along the beam path. For electrons or positrons the patterns may be detected with little background by detecting characteristic x-ray emission from a thin film as a function of the diffraction conditions of a collimated, ionizing, high energy beam. Uses of the effect for structure determination and atomic-resolution lensless imaging are suggested. PMID- 11415289 TI - Absence of an abrupt phase change from polycrystalline to amorphous in silicon with deposition temperature. AB - Using fluctuation electron microscopy, we have observed an increase in the mesoscopic spatial fluctuations in the diffracted intensity from vapor-deposited silicon thin films as a function of substrate temperature from the amorphous to polycrystalline regimes. We interpret this increase as an increase in paracrystalline medium-range order in the sample. A paracrystal consists of topologically crystalline grains in a disordered matrix; in this model the increase in ordering is caused by an increase in the grain size or density. Our observations are counter to the previous belief that the amorphous to polycrystalline transition is a discontinuous disorder-order phase transition. PMID- 11415290 TI - Entropically favored ordering: the metallurgy of Al(2)Cu revisited. AB - The famous sequence of precipitates which form upon heat treating Al-Cu is part of nearly every metallurgical textbook. Numerous precipitation (and other) experiments have led to a long-standing belief that the energetic ground state of Al(2)Cu is the theta phase. Modern first-principles calculations at T = 0 K surprisingly predict the energy of the observed Al(2)Cu-theta phase to be higher than that of its metastable counterpart, theta('). We show that vibrational entropy reverses this energetic preference at T approximately 150--200 degrees C, resolves the apparent discrepancy between theory and experiment, and hence plays a critical (but previously unsuspected) role in the precipitation sequence. PMID- 11415291 TI - E' centers in amorphous SiO(2) revisited: a new look at an old problem. AB - We present theoretical evidence that the paramagnetic E' defect centers in amorphous silicon dioxide (a-SiO(2)) do not have the same microscopic structures as those well-defined in the corresponding crystalline counterparts such as alpha quartz. We then present alternative models of some paramagnetic defects that account for the underlying experimental features of the E'-center variants in a SiO(2). We suggest that our new model should take the place of the conventional defect model of a-SiO(2). PMID- 11415292 TI - Microscopic theory of heterogeneity and nonexponential relaxations in supercooled liquids. AB - Recent experiments show that supercooled liquids around the glass transition temperature are "dynamically heterogeneous" [H. Sillescu, J. Non-Cryst. Solids 243, 81 (1999)]. Such heterogeneity is expected from the random first order transition theory of the glass transition. Using a microscopic approach based on this theory, we derive a relation between the departure from Debye relaxation as characterized by the beta value of a stretched exponential response function, phi(t) = e(-(t/tau(KWW))beta), and the fragility of the liquid. The beta value is also predicted to depend on temperature and to vanish as the ideal glass transition is approached at the Kauzmann temperature. PMID- 11415293 TI - Method for computing the anisotropy of the solid-liquid interfacial free energy. AB - We present a method to compute accurately the weak anisotropy of the solid-liquid interfacial free energy, a parameter which influences dendritic evolution in materials with atomically rough interfaces. The method is based on monitoring interfacial fluctuations during molecular dynamics simulation and extracting the interfacial stiffness which is an order of magnitude more anisotropic than the interfacial free energy. We present results for pure Ni with interatomic potentials derived from the embedded atom method. PMID- 11415294 TI - Dewetting patterns and molecular forces: a reconciliation. AB - We studied the dewetting of thin liquid polymer films from solid surfaces. Our experimental results lead to a consistent picture demonstrating the interplay between short- and long-range interfacial forces. Observations comprise nucleation and spinodal dewetting, as well as thermal nucleation of holes. The effective interface potential of the system, as reconstructed from the morphology of the dewetting patterns, agrees quantitatively with what is computed from the optical properties of the system. This shows that the assumption of additivity of dispersion potentials in multilayer systems yields good results. PMID- 11415295 TI - Unstable step meandering with elastic interactions. AB - We report on theoretical investigations of the influence of step interactions due to elasticity on unstable step meandering during molecular beam homoepitaxy. It is shown that elasticity causes coarsening of the cellular structure of the meander found in a previous work. The time dependence of step roughness is found to be robust, behaving as t(1/2). The lateral length scale coarsening is shown to be sensitive to the underlying physical mechanisms. In particular, the typical length follows the law t(alpha), with alpha = 1/6 or 1/4 depending on whether line diffusion is negligibly small or not. PMID- 11415296 TI - Simulations of atomic level stresses in systems of buried Ge /Si islands. AB - Stress distribution in laterally ordered arrays of coherent Ge islands on Si(001) buried in Si cap layers is examined using atomistic simulations. The obtained hydrostatic stress dependence on the spacer layer thickness shows a nearly linear inverse dependence, unlike the commonly used inverse cubic dependence derived in the framework of an isolated embedded force dipole source model. Additionally, the hydrostatic stress on the spacer surface is found to scale more closely with the area of the island rather than its volume as implicit in the use of the force dipole model. PMID- 11415297 TI - Nonuniqueness of the potentials of spin-density-functional theory. AB - It is shown that, contrary to widely held beliefs, the potentials of spin-density functional theory (SDFT) are not unique functionals of the spin densities. Explicit examples of distinct sets of potentials with the same ground-state densities are constructed. These findings imply that the zero-temperature exchange-correlation energy is not always a differentiable functional of the spin density. As a consequence, various types of applications of SDFT must be critically reexamined. PMID- 11415298 TI - Bilayer splitting in the electronic structure of heavily overdoped Bi(2)Sr(2)CaCu(2)O(8+delta). AB - The electronic structure of heavily overdoped Bi(2)Sr(2)CaCu(2)O(8+delta) is investigated by angle-resolved photoemission spectroscopy. The long-sought bilayer band splitting in this two-plane system is observed in both normal and superconducting states, which qualitatively agrees with the bilayer Hubbard model calculations. The maximum bilayer energy splitting is about 88 meV for the normal state feature, while it is only about 20 meV for the superconducting peak. PMID- 11415299 TI - Spin fluctuations in a magnetically frustrated metal LiV(2)O(4). AB - Inelastic neutron scattering is used to characterize spin fluctuations in the d electron heavy fermion spinel LiV(2)O(4). The spin-relaxation rate, gamma(Q), for Q = 0.6 A(-1) is 1.4(2) meV at low temperatures and increases linearly with temperature at a rate of 0.46(8)k(B). There is antiferromagnetic short-range order at low temperatures with a characteristic wave vector Q(c) = 0.64(2) A(-1) and a correlation length of 6(1) A. While warming shifts intensity towards lower Q, the staggered susceptibility peaks at a finite wave vector for T < 80 K. The data are compared with conventional heavy fermion systems, geometrically frustrated insulating magnets, and recent theories for LiV(2)O(4). PMID- 11415300 TI - Mesoscopic Kondo screening effect in a single-electron transistor embedded in a metallic ring. AB - We study the Kondo screening effect generated by a single-electron transistor or quantum dot embedded in a small metallic ring. When the ring circumference L becomes comparable to the fundamental length scale xi(0)(K) = Planck's constant over upsilon(F)/T(0)(K) associated with the bulk Kondo temperature, the Kondo resonance is strongly affected, depending on the total number of electrons (mod4) and magnetic flux threading the ring. The resulting Kondo-assisted persistent currents are also calculated in both Kondo and mixed-valence regimes, and the maximum values are found in the crossover region. PMID- 11415301 TI - Giant Hall effect in nonmagnetic granular metal films. AB - Nearly 3 orders of magnitude enhancement in the Hall coefficient is observed in Cu(x)-(SiO(2))(1--x) granular films. This large enhancement of the Hall coefficient not only is significantly larger than the prediction of the classical percolation theory, but also occurs at a metal concentration identified to be the quantum percolation threshold. Measurements of the electron dephasing length and magnetoresistance, plus the TEM characterization of microstructures, yield a physical picture consistent with the mechanism of the local quantum interference effect. PMID- 11415302 TI - Quantization of the Hall conductivity well beyond the adiabatic limit in pulsed magnetic fields. AB - We measure the Hall conductivity, sigma(xy), on a Corbino geometry sample of a high-mobility AlGaAs/GaAs heterostructure in a pulsed magnetic field. At a bath temperature about 80 mK, we observe well expressed plateaux in sigma(xy) at integer filling factors. In the pulsed magnetic field, the Laughlin condition of the phase coherence of the electron wave functions is strongly violated and, hence, is not crucial for sigma(xy) quantization. PMID- 11415303 TI - Opto-thermionic refrigeration in semiconductor heterostructures. AB - Combining the ideas of laser cooling and thermionic cooling, we have proposed an opto-thermionic cooling process, and investigated its cooling effect caused by the light emission from a quantum well embedded into a semiconductor pn junction. For a GaAs/AlGaAs opto-thermionic refrigerator in which the Auger recombination is the major nonradiative process, cooling can be achieved in a finite range of bias voltage. Using the measured values of the Auger coefficient, our calculated cooling rate is at least several watts/cm(2). PMID- 11415304 TI - From favorable atomic configurations to supershell structures: a new interpretation of conductance histograms. AB - Simulated minimum cross-section histograms of breaking Al nanocontacts are produced using molecular dynamics. The results allow a new interpretation of the controverted conductance histogram peaks based on preferential geometrical arrangements of nanocontact necks. As temperature increases, lower conductance peaks decrease in favor of broader and higher conductance structures. This reveals the existence of shell and supershell structures favored by the increased mobility of Al atoms. PMID- 11415305 TI - Dual nature of the electronic structure of (La(2--x--y)Nd(y)Sr(x))CuO(4) and La(1.85)Sr(0.15)CuO(4). AB - High resolution angle-resolved photoemission measurements have been carried out on (La(1.4--x)-Nd(0.6)Sr(x))CuO(4), a model system with static one-dimensional (1D) charge ordering (stripe), and (La(1.85)-Sr(0.15))CuO(4), a high temperature superconductor (T(c) = 40 K) with possible dynamic stripes. In addition to the straight segments near ( pi,0) and ( 0,pi) antinodal regions, we have identified the existence of spectral weight along the [1,1] nodal direction in the electronic structure of both systems. This observation of nodal state, together with the straight segments near antinodal regions, reveals the dual nature of the electronic structure of stripes due to the competition of order and disorder. PMID- 11415306 TI - Tunneling spectroscopy in small grains of superconducting MgB(2). AB - We report on tunneling spectroscopy experiments in small grains of the new binary intermetallic superconductor MgB(2). Experiments have been performed at 2.5 K using a low temperature scanning tunneling microscope. Good fit to the BCS model is obtained, with a gap value of 2 meV. In the framework of this model, this value should correspond to a surface critical temperature of 13.2 K. No evidence of gap anisotropy has been found. PMID- 11415307 TI - Determination of the spin polarization of half-metallic CrO(2) by point contact Andreev reflection. AB - Andreev reflection at a Pb/CrO(2) point contact has been used to determine the spin polarization of single-crystal CrO(2) films made by chemical vapor deposition. The spin polarization is found to be 0.96 +/- 0.01, which confirms that CrO(2) is a half-metallic ferromagnet, as theoretically predicted. PMID- 11415308 TI - Study of the e(g) orbitals in the bilayer manganite La(2--2x)Sr(1+2x)Mn(2)O(7) by using magnetic Compton-profile measurement. AB - From the magnetic Compton-profile (MCP) measurement, we have directly differentiated for the first time the populations in two e(g)-type orbitals ( x(2) - y(2) and 3z(2) - r(2)) in a manganite. The experimental MCP's along the [001] direction for La(2--2x)Sr(1+2x)Mn(2)O(7) at x = 0.35 and 0.42 are fitted by the theoretical profiles obtained from the (MnO(6))(8-) ab initio calculations. The calculation confirms that the MCP clearly detects the oxygen hybridization in the e(g) orbitals. The e(g) state is dominated by the x(2) - y(2)-type orbital with almost constant population, while the population in the 3z(2) - r(2)-type orbital decreases with increasing the hole concentration x. PMID- 11415309 TI - Ferromagnetism in magnetically doped III-V semiconductors. AB - The origin of ferromagnetism in semimagnetic III-V materials is discussed. The indirect exchange interaction caused by virtual electron excitations from magnetic impurity acceptor levels to the valence band can explain ferromagnetism in GaAs(Mn) in both degenerate and nondegenerate samples. Formation of ferromagnetic clusters and the percolation picture of phase transition describes well all available experimental data and allows us to predict the Mn-composition dependence of transition temperature in wurtzite (Ga,In,Al)N epitaxial layers. PMID- 11415310 TI - Perpendicular spin orientation in ultrasmall Fe islands on W(110). AB - We have studied the magnetism of Ag-coated Fe islands on W(110) by nuclear resonant scattering of synchrotron radiation at the 14.4 keV resonance of (57)Fe. Separated islands with an average diameter of 2.0 nm and monolayer thickness are formed at a Fe coverage of theta = 0.57 bulk monolayers. Time spectra of the nuclear decay were measured in the temperature range from 4.5 to 300 K. We find strong evidence for perpendicular spin orientation, which most likely results from the interplay of shape anisotropy and elastic strain in the islands. PMID- 11415311 TI - Optical resonance in a narrow slit in a thick metallic screen. AB - Interaction of TM-polarized waves with a subwavelength thick metallic slit has been analyzed. A Fabry-Perot-like behavior is reported. The resonance peaks, however, have very low magnitude and a systematic shift towards longer wavelengths is observed. The slit being narrow, this shift can be interpreted as the result of an aperture effect. Spectral transmission from a periodic array of such slits features the same peaks with a high increase in their magnitude, confirming that a grating acts as an amplifier of those resonances. Such a mechanism might explain the enhancement of the transmission observed in recent experiments [T. W. Ebbesen, H. J. Lezec, H. F. Ghaemi, T. Thio, and P. A. Wolff, Nature (London) 391, 667 (1998)]. PMID- 11415312 TI - Control of coherent acoustic phonons in semiconductor quantum wells. AB - Using subpicosecond optical pump-probe techniques, coherent zone-folded longitudinal acoustic phonons (ZFLAPs) were investigated in an InGaN multiple quantum well structure. A two-pump differential transmission technique was used to generate and control coherent ZFLAP oscillations through the relative timing and amplitude of the two pump pulses. Enhancement and suppression of ZFLAP oscillations were demonstrated, including complete cancellation of generated acoustic phonons for the first time in any material system. Coherent control was used to demonstrate that ZFLAPs are generated differently in InGaN multiple quantum wells than in GaAs/AlAs superlattices. PMID- 11415313 TI - Stimulated scattering of indirect excitons in coupled quantum wells: signature of a degenerate Bose-gas of excitons. AB - We observe and analyze strongly nonlinear photoluminescence kinetics of indirect excitons in GaAs/AlGaAs coupled quantum wells at low bath temperatures, > or = 50 mK. The long recombination lifetime of indirect excitons promotes accumulation of these Bose particles in the lowest energy states and allows the photoexcited excitons to cool down to temperatures where the dilute 2D gas of indirect excitons becomes statistically degenerate. Our main result--a strong enhancement of the exciton scattering rate to the low-energy states with increasing concentration of the indirect excitons--reveals bosonic stimulation of exciton scattering, which is a signature of a degenerate Bose-gas of excitons. PMID- 11415314 TI - Doppler broadening of in-flight positron annihilation radiation due to electron momentum. AB - We report the first observation of electron momentum contributions to the Doppler broadening of radiation produced by in-flight two-photon annihilation in solids. In these experiments an approximately 2.5 MeV positron beam impinged on thin polyethylene, aluminum, and gold targets. Since energetic positrons easily penetrate the nuclear Coulomb potential and do not cause a strong charge polarization, the experimental annihilation line shapes agree well with calculations based on a simple independent-particle model. Moreover, annihilations with the deepest core electrons are greatly enhanced. PMID- 11415315 TI - Energy transport induced by an external alternating field in strongly disordered media. AB - The delocalization of excitations in an ensemble of two-level systems with a strong disorder due to an external alternating acoustic or electric field is considered. The propagating modes are shown to appear if a ratio of the field amplitude to the frequency is large enough. Two complementary approaches, the static one similar to that of Anderson and the dynamic one related to Landau Zener, are developed. It is shown that the field-induced relaxation mechanism can have a strong influence on the kinetics. The internal friction is argued to be vastly affected by the relaxation mechanism proposed. PMID- 11415316 TI - New high-efficiency source of photon pairs for engineering quantum entanglement. AB - We constructed an efficient source of photon pairs using a waveguide-type nonlinear device and performed a two-photon interference experiment with an unbalanced Michelson interferometer. As the interferometer has two arms of different lengths, photons from the short arm arrive at the detector earlier than those from the long arm. We find that the arrival time difference (Delta L/c) and the time window of the coincidence counter (Delta T) are important parameters which determine the boundary between the classical and quantum regimes. Fringes of high visibility ( 80% +/- 10%) were observed when Delta T < Delta L/c. This result is explained only by quantum theory and is clear evidence for quantum entanglement of the interferometer's optical paths. PMID- 11415317 TI - Facilitated substrate transport through membrane proteins. AB - Substrate transport through the membrane protein maltoporin is facilitated by an affinity site in the channel. The analysis of the ion current fluctuations induced by penetration of the sugar into the channel yields the kinetic constants. Modification of the affinity site by replacing the aromatic residues suggests that nature has optimized the channel protein for maximum affinity at the extracellular side, as well as for an increased off-rate to eject a sugar trapped in the pore towards the periplasmic side. PMID- 11415318 TI - Photonic-band-gap resonator gyrotron. AB - We report the design and experimental demonstration of a gyrotron oscillator using a photonic-band-gap (PBG) structure to eliminate mode competition in a highly overmoded resonator. The PBG cavity supports a TE(041)-like mode at 140 GHz and is designed to have no competing modes over a minimum frequency range delta omega/omega of 30% about the design mode. Experimental operation of a PBG gyrotron at 68 kV and 5 A produced 25 kW of peak power in the design mode. No other modes were observed over the full predicted operating range about the design mode. PBG cavities show great promise for applications in vacuum electron devices in the millimeter- and submillimeter-wave bands. PMID- 11415319 TI - Bose-Einstein condensation in complex networks. AB - The evolution of many complex systems, including the World Wide Web, business, and citation networks, is encoded in the dynamic web describing the interactions between the system's constituents. Despite their irreversible and nonequilibrium nature these networks follow Bose statistics and can undergo Bose-Einstein condensation. Addressing the dynamical properties of these nonequilibrium systems within the framework of equilibrium quantum gases predicts that the "first-mover advantage," "fit-get-rich," and "winner-takes-all" phenomena observed in competitive systems are thermodynamically distinct phases of the underlying evolving networks. PMID- 11415320 TI - Comment on "Theory of diluted magnetic semiconductor ferromagnetism". PMID- 11415322 TI - Tapping spin glasses and ferromagnets on random graphs. AB - We consider a tapping dynamics, analogous to that in experiments on granular media, on spin glasses and ferromagnets on random thin graphs. Between taps, zero temperature single spin flip dynamics takes the system to a metastable state. Tapping corresponds to flipping simultaneously any spin with probability p. This dynamics leads to a stationary regime with a steady state energy E(p). We analytically solve this dynamics for the one-dimensional ferromagnet and +/-J spin glass. Numerical simulations for spin glasses and ferromagnets of higher connectivity are carried out; in particular, we find a novel first order transition for the ferromagnetic systems. PMID- 11415323 TI - Nonequilibrium model for the contact process in an ensemble of constant particle number. AB - We introduce and analyze numerically a nonequilibrium model with a conserved dynamics which is a realization of the contact process in an ensemble of constant particle number. The model possesses just one process in which particles jump around landing only on empty sites next to an existing particle. Particles are not allowed to land on a vacant site surrounded by empty sites. In contrast with the ordinary contact process, the present model does not have an absorbing state. In spite of lacking an absorbing state, the model displays properties that, in the thermodynamic limit, are identical to those of the ordinary contact process. PMID- 11415324 TI - Neutron star structure and the neutron radius of 208Pb. AB - We study relationships between the neutron-rich skin of a heavy nucleus and the properties of neutron-star crusts. Relativistic effective field theories with a thicker neutron skin in 208Pb have a larger electron fraction and a lower liquid to-solid transition density for neutron-rich matter. These properties are determined by the density dependence of the symmetry energy which we vary by adding nonlinear couplings between isoscalar and isovector mesons. An accurate measurement of the neutron radius in 208Pb-via parity violating electron scattering-may have important implications for the structure of the crust of neutron stars. PMID- 11415325 TI - Solar 8B and hep neutrino measurements from 1258 days of Super-Kamiokande data. AB - Solar neutrino measurements from 1258 days of data from the Super-Kamiokande detector are presented. The measurements are based on recoil electrons in the energy range 5.0-20.0 MeV. The measured solar neutrino flux is 2.32+/ 0.03(stat)+0.08-0.07(syst)x10(6) cm(-2) x s(-1), which is 45.1+/-0.5(stat)+1.6 1.4(syst)% of that predicted by the BP2000 SSM. The day vs night flux asymmetry (Phi(n)-Phi(d))/Phi(average) is 0.033+/-0.022(stat)+0.013-0.012(syst). The recoil electron energy spectrum is consistent with no spectral distortion. For the hep neutrino flux, we set a 90% C.L. upper limit of 40x10(3) cm(-2) x s(-1), which is 4.3 times the BP2000 SSM prediction. PMID- 11415326 TI - Constraints on neutrino oscillations using 1258 days of Super-Kamiokande solar neutrino data. AB - We report the result of a search for neutrino oscillations using precise measurements of the recoil electron energy spectrum and zenith angle variations of the solar neutrino flux from 1258 days of neutrino-electron scattering data in Super-Kamiokande. The absence of significant zenith angle variation and spectrum distortion places strong constraints on neutrino mixing and mass difference in a flux-independent way. Using the Super-Kamiokande flux measurement in addition, two allowed regions at large mixing are found. PMID- 11415328 TI - Sharpening low-energy, standard-model tests via correlation coefficients in neutron beta decay. AB - The correlation coefficients a, A, and B in neutron beta decay are proportional to the ratio of the axial-vector-to-vector weak coupling constants, gA/gV, to leading recoil order. With the advent of the next generation of neutron-decay experiments, the recoil-order corrections to these expressions become experimentally accessible, admitting a plurality of standard model (SM) tests. The measurement of both a and A, e.g., allows one to test the conserved-vector current (CVC) hypothesis and to search for second-class currents (SCC) independently. The anticipated precision of these measurements suggests that the bounds on CVC violation and SCC from studies of nuclear beta decay can be qualitatively bettered. PMID- 11415327 TI - Bounds on the CP asymmetry in b --> sgamma decays. AB - We have measured the CP asymmetry A(CP) identical with[gamma(b-->sgamma)-gammab- >sgamma)]/[gamma(b-->sgamma)+gamma(b-->sgamma)] to be A(CP) = (-0.079+/-0.108+/ 0.022) (1.0+/-0.030), implying that, at 90% confidence level, A(CP) lies between 0.27 and +0.10. These limits rule out some extreme non-standard-model predictions, but are consistent with most, as well as with the standard model. PMID- 11415330 TI - Breit-Pauli and direct perturbation theory calculations of relativistic helium polarizability. AB - Large Gaussian-type geminal wave function expansions and direct perturbation theory (DPT) of relativistic effects have been applied to calculate the relativistic contribution to the static dipole polarizability of the helium atom. It has been demonstrated that DPT is superior for this purpose to traditional Breit-Pauli calculations. The resulting value of the molar polarizability of 4He is 0.517254(1) cm3 x mol(-1), including a literature estimate of QED effects. As a by-product, a very accurate value of the nonrelativistic helium second hyperpolarizability, gamma = 43.104227(1) atomic units (without the mass polarization correction), has been obtained. PMID- 11415329 TI - Dynamics of the 16O(e, e'p) reaction at high missing energies. AB - We measured the cross section and response functions for the quasielastic 16O(e,e'p) reaction for missing energies 25< or =E(m)< or =120 MeV at missing momenta P(m)< or =340 MeV/c. For 25 symmetrical tilt grain boundary (GB) close to the solubility limit in the Cu(Ag) solid solution has been observed by means of Monte Carlo simulations at T = 600 K. More precisely, a finite thickness film almost pure in Ag, separating the two initial Cu(Ag) grains, can be obtained from a critical intergranular germ induced by the strong segregation of Ag in the GB. As this film is actually a single crystal, this implies a complete rearrangement of the GB core structure. Thus the initial GB is replaced by two Cu(Ag)/Ag(Cu) interfaces. Evidence is presented for the increase of the film thickness when approaching the solubility limit, as expected in wetting phenomena. PMID- 11415346 TI - Brownian motion of 2D vacancy islands by adatom terrace diffusion. AB - We have studied the Brownian motion of two-dimensional (2D) vacancy islands on Ag(110) at temperatures between 175 and 215 K. While the detachment of adatoms from the island and their diffusion on the terrace are permitted in this temperature range, the periphery diffusion of single adatoms is prohibited. The present scanning tunneling microscopy results provide the first direct experimental proof that the Brownian motion of the islands follows a simple scaling law with terrace diffusion being the rate limiting process. The activation energy of the vacancy island motion is determined to 0.41 eV. PMID- 11415347 TI - Theory for phase transitions in insulating V2O3. AB - We show that the recently proposed S = 2 bond model with orbital degrees of freedom for insulating V2O3 not only explains the anomalous magnetic ordering but also other mysteries of the magnetic phase transition. The model contains an additional orbital degree of freedom that exhibits a zero temperature quantum phase transition in the Ising universality class. PMID- 11415348 TI - Dispersion and damping of a two-dimensional plasmon in a metallic surface-state band. AB - We have studied, for the first time, the energy and the linewidth dispersion of a plasmon in a dense two-dimensional electron system in a metallic surface-state band on a silicon surface. As expected from the considerably high effective density and long Fermi wavelength of the system, the plasmon energy dispersion exhibited an excellent agreement with the nearly free-electron theory. However, in a small wave number region below the Landau edge, we have observed an anomalous linewidth dispersion which nearly free-electron theories do not predict. PMID- 11415349 TI - Imaging and spectroscopy of artificial-atom states in core/shell nanocrystal quantum dots. AB - Current imaging scanning tunneling microscopy is used to observe the electronic wave functions in InAs/ZnSe core/shell nanocrystals. Images taken at a bias corresponding to the s conduction band state show that it is localized in the central core region, while images at higher bias probing the p state reveal that it extends to the shell. This is supported by optical and tunneling spectroscopy data demonstrating that the s-p gap closes upon shell growth. Shapes of the current images resemble atomlike envelope wave functions of the quantum dot calculated within a particle in a box model. PMID- 11415350 TI - Ground-state phase diagram of 2D electrons in a high Landau level: a density matrix renormalization group study. AB - The ground-state phase diagram of 2D electrons in a high Landau level (index N = 2) is studied by the density-matrix renormalization group method. Pair correlation functions are systematically calculated for various filling factors from nu = 1/8 to 1/2. It is shown that the ground-state phase diagram consists of three different charge density wave states called stripe phase, bubble phase, and Wigner crystal. The boundary between the stripe and the bubble phases is determined to be nu(s-b)c approximately 0.38, and that for the bubble phase and Wigner crystal is nu(b-W)c approximately 0.24. Each transition is of first order. PMID- 11415351 TI - Integrable model for interacting electrons in metallic grains. AB - We find an integrable generalization of the BCS model with nonuniform Coulomb and pairing interaction. The Hamiltonian is integrable by construction since it is a functional of commuting operators; these operators, which therefore are constants of motion of the model, contain the anisotropic Gaudin Hamiltonians. The exact solution is obtained diagonalizing them by means of Bethe ansatz. Uniform pairing and Coulomb interaction are obtained as the "isotropic limit" of the Gaudin Hamiltonians. We discuss possible applications of this model to a single grain and to a system of few interacting grains. PMID- 11415352 TI - Closing the pseudogap by Zeeman splitting in Bi2Sr2CaCu2O8+y at high magnetic fields. AB - Interlayer tunneling resistivity is used to probe the low-energy density-of states (DOS) depletion due to the pseudogap in the normal state of Bi2Sr2CaCu2O8+y. Measurements up to 60 T reveal that a field that restores DOS to its ungapped state shows strikingly different temperature and doping dependencies from the characteristic fields of the superconducting state. The pseudogap closing field and the pseudogap temperature T small star, filled evaluated independently are related through a simple Zeeman energy scaling. These findings indicate a predominant role of spins over the orbital effects in the formation of the pseudogap. PMID- 11415353 TI - Nmr evidence for coexistence of superconductivity and ferromagnetic component in magnetic superconductor RuSr2YCu2O8: 99,101Ru and 63Cu NMR. AB - From Ru- and Cu-NMR studies, we present evidence for coexistence of superconductivity and ferromagnetism in a cuprate superconductor RuSr2YCu2O8 (RuY1212). The observation of a large enhancement of a radio-frequency field for the Ru-NMR signal at zero field reveals the existence of a ferromagnetic (FM) component in the ordered RuO2 plane below a Curie temperature of TM = 150 K. Just below the onset temperature of superconductivity T(onset)c = 45 K, a remarkable decrease of the nuclear spin-lattice relaxation rate 1/T1 was observed within the ordered RuO2 plane as well as the CuO2 plane, revealing that the superconducting gap coexists with the FM component in the RuO2 plane on a microscopic scale. In addition, from the observation of a sharp peak in 101(1/T1) at T(zero)c approximately 23 K where the resistivity becomes zero, we suggest that the motion of self-induced vortices originating from fluctuations of the FM component induces the resistivity between T(onset)c and T(zero)c in RuY1212. PMID- 11415354 TI - Phonon dispersion and electron-phonon coupling in MgB2 and AlB2. AB - We present a first principles investigation of the lattice dynamics and electron phonon coupling of the superconductor MgB2 and the isostructural AlB2 within the framework of density functional perturbation theory using a mixed-basis pseudopotential method. Complete phonon dispersion curves and Eliashberg functions alpha2F are calculated for both systems. The main differences are related to high frequency in-plane boron vibrations, which are strongly softened in MgB2 and exhibit an exceptionally strong electron-phonon coupling. We also report on Raman measurements, which support the theoretical findings. Implications for the superconducting transition temperature are briefly discussed. PMID- 11415355 TI - Unconventional superconductivity in electron-doped layered Li0.48(THF)yHfNCL. AB - We report magnetic susceptibility measurements on a layered superconductor Li0.48(THF)0.3HfNCl having Tc approximately 26 K. The present study revealed that (a) the Fermi level density of states is small, N*(EF) approximately 0.25 states/(eV spin f.u.), (b) mass enhancement is negligible, gamma; approximately 1, (c) electron-phonon coupling is weak, lambda(ep)<<1, (d) exchange enhancement is negligible, 1/(1+F(a)0) approximately 1, and (e) electronic density parameter is large, r(2D)s approximately 10.3 (i.e., low-carrier density). It is difficult to explain the origin of the high Tc in terms of the conventional phonon (BCS) mechanism of superconductivity. PMID- 11415356 TI - Exchange field induced magnetoresistance in colossal magnetoresistance manganites. AB - The effect of an exchange field on the electrical transport in thin films of metallic ferromagnetic manganites has been investigated. The exchange field was induced both by direct exchange coupling in a ferromagnet/antiferromagnet multilayer and by indirect exchange interaction in a ferromagnet/paramagnet metallic superlattice. The electrical resistance of the metallic manganite layers was found to be determined by the magnitude of the vector sum of the effective exchange field and the external magnetic field. PMID- 11415357 TI - Magnetic ordering and phase transition in MnO embedded in a porous glass. AB - We present the results of a neutron diffraction study of the antiferromagnet MnO embedded in a porous glass. The type of magnetic ordering and the structural distortion are similar to those of the bulk, but the ordered magnetic moment of 3.84(4)muB/ion is strongly reduced and the Neel temperature is enhanced. The magnetic transition is second order, in contrast to the first order transition of the bulk. The size of the magnetic region is smaller than the average size of the nanoparticles. The reasons for this behavior are discussed. PMID- 11415358 TI - Experimental evidence for electron channeling in Fe /Aau (100) superlattices. AB - We present transport and structural data from epitaxial (100) and (111) Au/Fe superlattices grown by molecular beam epitaxy. From their analysis, we conclude that an electron channeling mechanism, due to strong specular reflection of the minority spin carrier at the Au/Fe interfaces, is responsible for the high conductivity in the (100) superlattices. PMID- 11415359 TI - Stimulated secondary emission from semiconductor microcavities. AB - We find strong influence of final-state stimulation on the time-resolved light emission dynamics from semiconductor microcavities after pulsed excitation allowing angle-resonant polariton-polariton scattering on the lower-polariton branch. The polariton dynamics can be controlled by injection of final-state polaritons at densities below a polariton saturation density of 5x10(8) cm(-2). A bosonic enhancement factor in the dynamics of up to 700 is evaluated. PMID- 11415360 TI - Electron-hole droplet formation in direct-gap semiconductors observed by mid infrared pump-probe spectroscopy. AB - Mid-infrared pump-probe measurements with subpicosecond time resolution reveal the existence of a metastable condensed phase of the electron-hole ensemble in a direct-gap semiconductor CuCl. High-density electrons and holes are directly created in a low-temperature state by the resonant femtosecond excitation of excitons above the Mott transition density. Strong metallic reflection with a plasma frequency Planck's over 2pi(omega)p approximately 0.5 eV builds up within 0.3 ps. Within a few picoseconds, the mid-infrared reflection spectrum is transformed from metalliclike into colloidlike. The observed resonance feature at Planck's over 2pi(omega)p/sqrt[3] allows us to obtain the carrier density in the metastable electron-hole droplets of 2x10(20) cm(-3). PMID- 11415361 TI - Higher order ferroic switching induced by scanning force microscopy. AB - We present the observation of ferroelastoelectric switching in a ferroelectric material. It is achieved in barium titanate thin film by simultaneously applying electric field and compressive stress with the tip of a scanning force microscope. For low compressive stresses, the presented measurements reveal classical ferroelectric domain reversal, i.e., the spontaneous polarization is aligned parallel to the applied electric field. However, for high compressive stresses the direction of polarization after switching is antiparallel to the poling field, demonstrating ferroelastoelectric switching. PMID- 11415362 TI - Irreversibility in asymptotic manipulations of entanglement. AB - We show that the process of entanglement distillation is irreversible by showing that the entanglement cost of a bound entangled state is finite. Such irreversibility remains even if extra pure entanglement is loaned to assist the distillation process. PMID- 11415363 TI - Hiding bits in bell states. AB - We present a scheme for hiding bits in Bell states that is secure even when the sharers, Alice and Bob, are allowed to carry out local quantum operations and classical communication. We prove that the information that Alice and Bob can gain about a hidden bit is exponentially small in n, the number of qubits in each share, and can be made arbitrarily small for hiding multiple bits. We indicate an alternative efficient low-entanglement method for preparing the shared quantum states. We discuss how our scheme can be implemented using present-day quantum optics. PMID- 11415364 TI - Benchmarking quantum computers: the five-qubit error correcting code. AB - The smallest quantum code that can correct all one-qubit errors is based on five qubits. We experimentally implemented the encoding, decoding, and error correction quantum networks using nuclear magnetic resonance on a five spin subsystem of labeled crotonic acid. The ability to correct each error was verified by tomography of the process. The use of error correction for benchmarking quantum networks is discussed, and we infer that the fidelity achieved in our experiment is sufficient for preserving entanglement. PMID- 11415365 TI - New stopping criteria for segmenting DNA sequences. AB - We propose a solution on the stopping criterion in segmenting inhomogeneous DNA sequences with complex statistical patterns. This new stopping criterion is based on Bayesian information criterion in the model selection framework. When this criterion is applied to telomere of S. cerevisiae and the complete sequence of E. coli, borders of biologically meaningful units were identified, and a more reasonable number of domains was obtained. We also introduce a measure called segmentation strength which can be used to control the delineation of large domains. The relationship between the average domain size and the threshold of segmentation strength is determined for several genome sequences. PMID- 11415366 TI - Error threshold for spatially resolved evolution in the quasispecies model. AB - The error threshold for quasispecies in 1, 2, 3, and infinity dimensions is investigated by stochastic simulation and analytically. The results show a monotonic decrease in the maximal sustainable error probability with decreasing diffusion coefficient, independently of the spatial dimension. It is thereby established that physical interactions between sequences are necessary in order for spatial effects to enhance the stabilization of biological information. The analytically tractable behavior in an infinity-dimensional (simplex) space provides a good guide to the spatial dependence of the error threshold in lower dimensional Euclidean space. PMID- 11415367 TI - Temporal correlations and neural spike train entropy. AB - Sampling considerations limit the experimental conditions under which information theoretic analyses of neurophysiological data yield reliable results. We develop a procedure for computing the full temporal entropy and information of ensembles of neural spike trains, which performs reliably for limited samples of data. This approach also yields insight to the role of correlations between spikes in temporal coding mechanisms. The method, when applied to recordings from complex cells of the monkey primary visual cortex, results in lower rms error information estimates in comparison to a "brute force" approach. PMID- 11415368 TI - Quantitative in-line phase-contrast imaging with multienergy X rays. AB - We present a new method for quantitative nondestructive characterization of objects by x-ray phase-contrast imaging. Spatial distributions of the projected values of the complex refractive index in the sample are reconstructed by processing near-field images collected at a fixed sample-to-detector distance using a polychromatic incident beam and an energy-sensitive area detector, such as a CCD used in the photon-counting spectroscopy mode. The method has the potential advantages of decreased radiation dose and increased accuracy compared to conventional techniques of x-ray imaging. PMID- 11415369 TI - Freak waves in random oceanic sea states. AB - Freak waves are very large, rare events in a random ocean wave train. Here we study their generation in a random sea state characterized by the Joint North Sea Wave Project spectrum. We assume, to cubic order in nonlinearity, that the wave dynamics are governed by the nonlinear Schrodinger (NLS) equation. We show from extensive numerical simulations of the NLS equation how freak waves in a random sea state are more likely to occur for large values of the Phillips parameter alpha and the enhancement coefficient gamma. Comparison with linear simulations is also reported. PMID- 11415370 TI - Weighted evolving networks. AB - Many biological, ecological, and economic systems are best described by weighted networks, as the nodes interact with each other with varying strength. However, most evolving network models studied so far are binary, the link strength being either 0 or 1. In this paper we introduce and investigate the scaling properties of a class of models which assign weights to the links as the network evolves. The combined numerical and analytical approach indicates that asymptotically the total weight distribution converges to the scaling behavior of the connectivity distribution, but this convergence is hampered by strong logarithmic corrections. PMID- 11415371 TI - Comment on "nonstationarity induced by long-time noise correlations in the Langevin equation". PMID- 11415373 TI - Dependence of the fractional quantum Hall edge critical exponent on the range of interaction. PMID- 11415374 TI - Coulomb interaction and instability of CE-type structure in half-doped manganites. PMID- 11415376 TI - NMR analog of the quantum disentanglement eraser. AB - We report the implementation of a three-spin quantum disentanglement eraser on a liquid-state NMR quantum information processor. A key feature of this experiment was its use of pulsed magnetic field gradients to mimic projective measurements. This ability is an important step towards the development of an experimentally controllable system which can simulate any quantum dynamics, both coherent and decoherent. PMID- 11415377 TI - Electric dipole moment cancellations in D-brane models. AB - We analyze the possibility of simultaneous electron, neutron, and mercury electric dipole moment (EDM) cancellations in the mSUGRA and D-brane models. We find that the mercury EDM constraint practically rules out the cancellation scenario in D-brane models, whereas in the context of mSUGRA it is still allowed with some fine tuning. PMID- 11415378 TI - Upper limits on sparticle masses from g -- 2 and the possibility for discovery of supersymmetry at colliders and in dark matter searches. AB - We analyze the implications of the new physics effect seen in the g -- 2 Brookhaven measurement and show that if the effect arises from supersymmetry, then the sign of the Higgs mixing parameter mu is determined to be positive in the standard sign convention. Further, analyses within the minimal supergravity model show that the Brookhaven result leads to upper limits on the universal gaugino and scalar masses of m(1/2) < or = 800 GeV and m(0) < or = 1.5 TeV for tan beta < or = 55. Our analysis strongly suggests that supersymmetry via production of sparticles must be found at the CERN Large Hadron Collider. Further, sgn(mu) positive is favorable for the discovery of supersymmetric cold dark matter. PMID- 11415379 TI - Eta(') mass and chiral symmetry breaking at large N(c) and N(f). AB - We propose a method for implementing the large- N(c), large-N(f) limit of QCD at the effective Lagrangian level. Depending on the value of the ratio N(f)/N(c), different patterns of chiral symmetry breaking can arise, leading in particular to different behaviors of the eta(') mass in the combined large-N limit. PMID- 11415380 TI - Search for three-nucleon force effects in analyzing powers for p-->d elastic scattering. AB - A series of measurements have been performed at KVI to obtain the vector analyzing power A(y) of the (2)H(p-->,pd) reaction as a function of incident beam energy at energies of 120, 135, 150, and 170 MeV. For all these measurements, a range of theta(c.m.) from 30 degrees to 170 degrees has been covered. The purpose of these investigations is to observe possible spin-dependent effects beyond two nucleon forces. When compared to the predictions of Faddeev calculations, based on two-nucleon forces only, significant deviations are observed at all energies and at center-of-mass angles between 70 degrees and 130 degrees. The addition of present-day three-nucleon forces does not improve the description of the data, demonstrating the still insufficient understanding of the properties of three nucleon systems. PMID- 11415381 TI - Evidence for the wobbling mode in nuclei. AB - The nucleus (163)Lu has been populated through the fusion-evaporation reaction (139)La((29)Si,5n)(163)Lu with a beam energy of 152 MeV. The electromagnetic properties of several connecting transitions between two presumably triaxial, strongly deformed (TSD) bands have been studied. Evidence is presented for the assignment of the excited TSD band as a wobbling mode built on the yrast TSD band, based on comparisons to new calculations in which an aligned particle is coupled to a strongly deformed triaxial rotor. The wobbling mode is uniquely related to triaxiality in nuclei. PMID- 11415382 TI - Experimental demonstration of entanglement-enhanced rotation angle estimation using trapped ions. AB - We experimentally investigate three methods, utilizing different atomic observables and entangled states, to increase the sensitivity of rotation angle measurements beyond the "standard quantum limit" for nonentangled states. All methods use a form of quantum mechanical "squeezing." In a system of two entangled trapped (9)Be(+) ions we observe a reduction in uncertainty of rotation angle below the standard quantum limit for all three methods including all sources of noise. As an application, we demonstrate an increase in precision of frequency measurement in a Ramsey spectroscopy experiment. PMID- 11415383 TI - Near-field tomography without phase retrieval. AB - We investigate the near-field inverse scattering problem with evanescent waves. An analytic solution to this problem within the weak-scattering approximation is used to show that the usual Rayleigh limit may be overcome even when measurements are made without phase information. Applications to a novel form of three dimensional microscopy with subwavelength resolution are described. PMID- 11415384 TI - Local low dimensionality of atmospheric dynamics. AB - A statistic, the BV (bred vector) dimension, is introduced to measure the effective local finite-time dimensionality of a spatiotemporally chaotic system. It is shown that the Earth's atmosphere often has low BV dimension, and the implications for improving weather forecasting are discussed. PMID- 11415385 TI - Heat conduction in the disordered harmonic chain revisited. AB - A general formulation is developed to study heat conduction in disordered harmonic chains with arbitrary heat baths, satisfying the fluctuation-dissipation theorem. A simple formal expression for the heat current J is obtained, from which its asymptotic system-size (N) dependence is extracted. It is shown that "thermal conductivity" depends not just on the system itself but also on the spectral properties of the heat baths. As special cases we recover earlier results that gave J approximately 1/N(3/2) for fixed boundaries and J approximately 1/N(1/2) for free boundaries. Other choices give other power laws including the "Fourier behavior" J approximately 1/N. PMID- 11415386 TI - Longitudinal vortices in granular flows. AB - We present a new instability observed in rapid granular flows down rough inclined planes. For high inclinations and flow rates, the free surface of the flow experiences a regular deformation in the transverse direction. Measurements of the surface velocities imply that this instability is associated with the formation of longitudinal vortices in the granular flow. From the experimental observations, we propose a mechanism for the longitudinal vortex formation based on the concept of granular temperature. PMID- 11415387 TI - Critical geometry of two-dimensional passive scalar turbulence. AB - Passive scalars advected by a magnetically driven two-dimensional turbulent flow are analyzed using methods of statistical topography. The passive tracer concentration is interpreted as the height of a random surface, and the scaling properties of its contour loops are analyzed. Various exponents that describe the loop ensemble are measured and compared to a scaling theory. This leads to a geometrical criterion for the intermittency of scalar fluctuations. PMID- 11415388 TI - Spatiotemporal scaling of solar surface flows. AB - The sun provides an excellent natural laboratory for nonlinear phenomena. We use motions of magnetic bright points on the solar surface, at the smallest scales yet observed, to study the small scale dynamics of the photospheric plasma. The paths of the bright points are analyzed within a continuous time random walk framework. Their spatial and temporal scaling suggests that the observed motions are the walks of imperfectly correlated tracers on a turbulent fluid flow in the lanes between granular convection cells. PMID- 11415389 TI - Domain coarsening in electroconvection. AB - We report on experimental measurements of the growth of regular domains evolving from an irregular pattern in electroconvection. The late-time growth of the domains is consistent with the size of the domains scaling as t(n). We use two isotropic measurements of the domain size: the structure factor and the domain wall length. Measurements using the structure factor are consistent with t(1/5) growth. Measurements using the domain wall length are consistent with t(1/4) growth. One source of this discrepancy is the fact that the distribution of local wave numbers is approximately independent of the domain size. In addition, we measure the anisotropy of the growing domains. PMID- 11415390 TI - Characterization of a high-gain harmonic-generation free-electron laser at saturation. AB - We report on an experimental investigation characterizing the output of a high gain harmonic-generation (HGHG) free-electron laser (FEL) at saturation. A seed CO2 laser at a wavelength of 10.6 microm was used to generate amplified FEL output at 5.3 microm. Measurement of the frequency spectrum, pulse duration, and correlation length of the 5.3 microm output verified that the light is longitudinally coherent. Investigation of the electron energy distribution and output harmonic energies provides evidence for saturated HGHG FEL operation. PMID- 11415391 TI - Observation of sub-Poisson fluctuations in the intensity of the seventh coherent spontaneous harmonic emitted by a rf linac free-electron laser. AB - We report the observation of sub-Poisson intensity fluctuations in the coherent spontaneous harmonic radiation generated by an infrared free-electron laser in a photon counting experiment using a well-defined ensemble of electron pulses. These observations constitute the first observation of a nonclassical state of the radiation field generated by a beam of free electrons. The fluctuations observed in the experiment are smaller than those expected from semiclassical radiation theory, and larger than those expected from electron shot noise. PMID- 11415392 TI - Capillary condensation of a nematic liquid crystal observed by force spectroscopy. AB - We have observed capillary condensation in nanometer-thin films of nematic liquid crystals using force spectroscopy. The liquid crystal was confined to a submicron gap between a flat substrate and a microsphere, attached to the cantilever of an atomic force microscope. A long-ranged and strongly attractive force due to capillary condensation of a nematic phase was observed at temperatures close to the bulk isotropic-nematic phase transition. The critical point, terminating the first-order confined isotropic-nematic transition line, was determined for the first time. PMID- 11415393 TI - Liquidlike morphological transformations in monolamellar polymer crystals. AB - We performed annealing experiments on monolamellar polyethyleneoxid crystals. Even well below the melting point, local relaxations of metastable folded states led to pronounced liquidlike morphological changes on a micron scale of shape and perimeter of the crystalline domains, similar to dewetting of liquids. First, a rim of less folded molecules was formed around the lamellae, making relaxations of the interior difficult. On longer time scales, hole formation within these domains was observed. Our experiments showed that, due to such spatial anisotropy in relaxations, polymer crystal morphology depends on the thermal pathway taken, implying that a time-temperature superposition principle does not apply. PMID- 11415394 TI - Phase diagram of carbon dioxide: evidence for a new associated phase. AB - The stability of CO(2) phases has been investigated up to 50 GPa and 750 K by in situ Raman spectroscopy and visual observations using externally heated diamond anvil cells. A new phase (CO(2)-II) exists above 20 GPa and 500 K, which can be quenched to ambient temperature. The vibrational spectrum of this new CO(2) polymorph suggests the dimeric pairing of molecules. Based on the present in situ data and previous laser-heating results, we present new constraints for the phase diagram of carbon dioxide to 50 GPa and 2000 K. We find that carbon dioxide exhibits dramatic changes, both in the molecular configuration and in the nature of intermolecular interaction at high pressures and temperatures. PMID- 11415395 TI - Reentrant behavior of relaxation time with viscosity at varying composition in binary mixtures. AB - In order to understand the long known anomalies in the composition dependence of diffusion and viscosity of binary mixtures, we introduce here two new models and carry out extensive molecular dynamics simulations. In these models, the two molecular species (A and B) have the same diameter and mass. In model I the interspecies interaction is more attractive than that between the pure components, while the reverse is true for model II. Simulations and mode coupling theory calculations reveal that the models can capture a wide variety of behavior observed in experiments, including the reentrant viscosity dependence of relaxation time. PMID- 11415396 TI - Discontinuous bending rigidity and cosurfactant adsorption of amphiphile layers. AB - We study a simple model for an amphiphilic layer that may adsorb cosurfactant molecules from a reservoir. Upon varying the length of the cosurfactant, we obtain a discontinuous change of the adsorption rate and a corresponding discontinuity of the bending rigidity kappa. With realistic values for the chemical potential and the interaction energy of the hydrophobic tails, our model accounts quantitatively for the measured rigidity and the discontinuity observed for the ternary system AOT/water/oil and for SDS/alcohol bilayers. PMID- 11415397 TI - Coupled capillary wave fluctuations in thin aqueous films on an aqueous subphase. AB - X-ray scattering and interfacial tension measurements are used to demonstrate the formation of nanometer-thick aqueous films on aqueous bulk subphases from polymer salt biphase mixtures. X-ray scattering determines a coupling constant that characterizes the coupled capillary wave fluctuations of the liquid-vapor and liquid-liquid interfaces of this thin film. These data also determine an effective Hamakar constant that characterizes the long-range interaction between the interfaces and parameters that characterize the short-range part of the interfacial interaction. PMID- 11415398 TI - How asymmetric islands become symmetric. AB - Scanning tunneling microscopy reveals that sputtering of the Au(110) surface results in the formation of vacancy islands with a broken mirror symmetry. These islands exhibit two types of steps on opposite sides: a lower energy (111) step and a higher energy (331) step. We analyze the thermal fluctuations and especially the kink distribution of such vacancy islands. Despite the broken symmetry, which they adopt internally, these islands show a symmetric average outer contour. Their coarsening proceeds via a variety of pathways, often leading to new, symmetric structures, with exclusively (111) steps. The lowest energy vacancy configuration is a bound pair of two vacancy lines or islands. PMID- 11415399 TI - Coadsorption of CO and NO on the Pd(111) surface: combined ab initio and Monte Carlo study. AB - Adsorption and coadsorption of nitric oxide (NO) and carbon monoxide (CO) on the Pd(111) surface are studied by combining first principles (FP) calculations and Monte Carlo (MC) simulations. From FP adsorption energies and molecule-molecule interactions we construct an interaction model, which is used in MC. We do several simulations with different coverages and CO/NO ratios. The simulations provide 0.75 monolayer (ML) for a saturation coverage, which is in excellent agreement with experiments. The results indicate that at 0.75 ML coverage, NO molecules take over the hollow sites and push CO molecules mainly onto bridge sites. PMID- 11415400 TI - Atomic-scale dynamics of the formation and dissolution of carbon clusters in SiO(2). AB - Oxidation of SiC produces SiO(2) while CO is released. A "reoxidation" step at lower temperatures is, however, necessary to produce high-quality SiO(2). This step is believed to cleanse the oxide of residual C without further oxidation of the SiC substrate. We report first-principles calculations that describe the nucleation and growth of O-deficient C clusters in SiO(2) under oxidation conditions, fed by the production of CO at the advancing interface, and their gradual dissolution by the supply of O under reoxidation conditions. We predict that both CO and CO(2) are released during both steps. PMID- 11415401 TI - COI oxidation on a single Pd atom supported on magnesia. AB - The oxidation of CO on single Pd atoms anchored to MgO(100) surface oxygen vacancies is studied with temperature-programmed-reaction mass spectrometry and infrared spectroscopy. In one-heating-cycle experiments, CO(2), formed from O(2) and CO preadsorbed at 90 K, is detected at 260 and 500 K. Ab-initio simulations suggest two reaction routes, with Pd(CO)(2)O(2) and PdCO(3)CO found as precursors for the low and high temperature channels, respectively. Both reactions result in annealing of the vacancy and induce migration and coalescence of the remaining Pd CO to form larger clusters. PMID- 11415402 TI - Prediction of electronic excited states of adsorbates on metal surfaces from first principles. AB - We present the first ab initio prediction of localized electronic excited states in a periodically infinite condensed phase, a heretofore intractable goal. In particular, we examined local excitations within a CO molecule adsorbed on Pd(111). The calculation allows a configuration interaction treatment of a local region, while its interaction with the extended condensed phase is described via an embedding potential obtained from periodic density functional theory. Our work lays the foundation of a microscopic understanding of photochemistry and spectroscopy on metal surfaces. PMID- 11415403 TI - First principles molecular dynamics of dense plasmas. AB - Ab initio molecular dynamics calculations are performed for the equation of state of aluminum, spanning condensed matter and dense plasma regimes. Electronic exchange and correlation are included with either a zero- or finite-temperature local density approximation potential. Standard methods are extended to above the Fermi temperature by using final state pseudopotentials to describe thermally excited ion cores. The predicted Hugoniot equation of state agrees well with earlier plasma theories and with experiment for temperatures from 0 to 3 x 10(6) K. PMID- 11415404 TI - Excitonic effects on the silicon plasmon resonance. AB - We present an ab initio calculation of the electron energy loss spectrum of silicon including local-field, self-energy, and excitonic effects. When self energy corrections are added to the standard random phase approximation (RPA) the line shape of the plasmon resonance worsens. The electron-hole interaction cancels this correction and improves the result both compared to the RPA and to the self-energy one, yielding very good agreement between theory and experiment provided that the mixing of interband transitions of both positive and negative frequencies is included. PMID- 11415405 TI - Charge ordering and spin dynamics in NaV(2)O(5). AB - We report high-resolution neutron inelastic scattering experiments on the spin excitations of NaV(2)O(5). Below T(c), two branches with distinct energy gaps are identified. From the dispersion and intensity of the spin excitation modes, we deduce the precise zigzag charge distribution on the ladder rungs and the corresponding charge order: Delta(c) approximately 0.6. We argue that the spin gaps observed in the low-T phase of this compound are primarily due to the charge transfer. PMID- 11415406 TI - Electronic structure of carbon nanocones. AB - Topology related changes in the local density of states near the apex of carbon nanocones are investigated using both tight-binding and ab initio calculations. Sharp resonant states are found to dominate the electronic structure in the region close to the Fermi energy. The strength and the position of these states with respect to the Fermi level depend sensitively on the number and the relative positions of the pentagons constituting the conical tip. Carbon nanocones are thus proposed as good candidates for nanoprobes in scanning probe microscopy. PMID- 11415407 TI - Anisotropic superconducting properties of aligned MgB(2) crystallites. AB - Samples of aligned MgB(2) crystallites have been prepared, allowing for the first time the direct identification of an upper critical field anisotropy H(ab)(c2)/H(c)(c2) = xi(ab)/xi(c) approximately 1.7, with xi(o,ab) approximately 70 A, xi(o,c) approximately 40 A, and a mass anisotropy ratio m(ab)/m(c) approximately 0.3. A ferromagnetic background signal was identified, possibly related to the raw materials purity. PMID- 11415408 TI - Resonance peak in Sr(2)RuO(4): signature of spin triplet pairing. AB - We study the dynamical spin susceptibility, chi(q,omega), in the normal and superconducting states of Sr(2)RuO(4). In the normal state, we find a peak in the vicinity of Q(i) approximately (0.72 pi,0.72 pi) in agreement with recent inelastic neutron scattering experiments. We predict that for spin triplet pairing in the superconducting state a resonance peak appears in the out-of-plane component of chi, but is absent in the in-plane component. In contrast, no resonance peak is expected for spin singlet pairing. PMID- 11415409 TI - Nesting symmetries and diffusion in disordered d-wave superconductors. AB - The low-energy density of states (DOS) of disordered 2D d-wave superconductors is extremely sensitive to details of both the disorder model and the electronic band structure. Using diagrammatic methods and numerical solutions of the Bogoliubov- de Gennes equations, we show that the physical origin of this sensitivity is the existence of a novel diffusive mode with momentum close to (pi,pi) which is gapless only in systems with a global nesting symmetry. We find that in generic situations the DOS vanishes at the Fermi level. However, proximity to the highly symmetric case may nevertheless lead to observable nonmonotonic behavior of the DOS in the cuprates. PMID- 11415410 TI - Ultrasound attenuation in Sr(2)RuO(4): an angle-resolved study of the superconducting gap function. AB - We present a study of the electronic ultrasound attenuation alpha in the unconventional superconductor Sr(2)RuO(4). The power law behavior of alpha at temperatures down to T(c)/30 clearly indicates the presence of nodes in the gap. In the normal state, we find an enormous anisotropy of alpha in the basal plane of the tetragonal structure. In the superconducting state, the temperature dependence of alpha also exhibits significant anisotropy. We discuss these results in relation to possible gap functions. PMID- 11415411 TI - Interlayer transport in the highly anisotropic misfit-layer superconductor [(LaSe)(1.14)](NbSe(2)). AB - The interlayer transport in a two-dimensional superconductor can reveal a peak in the temperature as well as the magnetic field dependence of the resistivity near the superconducting transition. The experiment was performed on the highly anisotropic misfit-layer superconductor [(LaSe)(1.14)](NbSe(2)) with T(c) of 1.2 K. The effect is interpreted within the tunneling mechanism of the charge transport across the Josephson-coupled layers via two parallel channels--the quasiparticles and the Cooper pairs. Similar behavior can be found in the high T(c) cuprates but there it is inevitably interfering with the anomalous normal state. The upper critical magnetic field can be obtained from the interlayer tunneling conductance. PMID- 11415412 TI - Coupled magnetic excitations in single crystal PrBa(2)Cu(3)O(6.2). AB - The dispersion of the low-energy magnetic excitations of the Pr sublattice in PrBa(2)Cu(3)O(6.2) is determined by inelastic neutron scattering measurements on a single crystal. The dispersion, which shows the effect of interactions with the Cu spin waves, is well described by a model of the coupled Cu-Pr magnetic system. This model enables values for the principal exchange constants to be determined. The results suggest that both Pr-Pr and Cu-Pr interactions are important in producing the anomalously high ordering temperature of the Pr sublattice. Measurements of the Cu optic spin wave mode show that the interlayer Cu-Cu exchange is significantly lower than in YBa(2)Cu(3)O(6.2). PMID- 11415413 TI - Electroresistance and electronic phase separation in mixed-valent manganites. AB - The sensitivity of transport in colossal magnetoresistance (CMR) manganites to external electric and magnetic fields is examined using field effect configurations with La(0.7)Ca(0.3)MnO(3) (LCMO), Na(0.7)Sr(0.3)MnO(3), La(0.7)Ba(0.3)MnO(3), and La(0.5)Ca(0.5)MnO(3) (0.5-doped LCMO) channels, and ferroelectric PbZr(0.2)Ti(0.8)O(3) (PZT) or dielectric (SrTiO(3)) gates. A large electroresistance (ER) of approximately 76% at 4 x 10(5) V/cm is found in LCMO with PZT-ferroelectric gate, but the magnitude of the effect is much smaller (a few percent) in the other three channels. The ER and CMR effects are remarkably complimentary. The size and systematics of the effect strongly favor a percolative phase separation picture. PMID- 11415414 TI - Nonlinear magneto-optical properties of colossal magnetoresistive manganites. AB - Pr(1--x)CaxMnO(3) and Nd(1--x)SrxMnO(3) were investigated with three-photon difference frequency generation (DFG). This method allows one to determine both the crystalline and the magnetic symmetry. In the highly ordered low-temperature phase of Nd(0.50)Sr(0.50)MnO(3), a DFG contribution coupling simultaneously to antiferromagnetic and charge ordering was observed and used to reveal the formation of domains. Thus, magnetically induced three-photon processes are introduced into the fields of both nonlinear magneto-optics and colossal magnetoresistance as a powerful new method. PMID- 11415415 TI - Luminescent FeSi(2) crystal structures induced by heteroepitaxial stress on Si(111). AB - The crystal structures and the luminescent properties of FeSi(2) in the FeSi(2)/Si heteroepitaxial system have been investigated by first principles calculations. The results indicate that the heteroepitaxial beta-FeSi(2) facing Si(111) by the (110) plane will be deformed from an orthorhombic to a monoclinic P2(1)/c structure with a gamma angle of 95 degrees. The strained crystal has a direct gap band structure and a finite oscillator strength of 0.7 between the band edges at the Y point. Since an indirect type band structure is obtained for other heteroepitaxial relationships, as well as for the bulk beta-FeSi(2), we propose the strained FeSi(2)(110)/Si(111) structure to be the origin of the luminescence observed in the FeSi(2)/Si systems. PMID- 11415416 TI - Photoluminescence of isotopically purified silicon: how sharp are bound exciton transitions? AB - We report the first high resolution photoluminescence studies of isotopically pure Si (99.896% (28)Si). New information is obtained on isotopic effects on the indirect band gap energy, phonon energies, and phonon broadenings, which is in good agreement with calculations and previous results obtained in Ge and diamond. Remarkably, the linewidths of the no-phonon boron and phosphorus bound exciton transitions in the (28)Si sample are much narrower than in natural Si and are not well resolved at our maximum instrumental resolution of approximately 0.014 cm( 1). The removal of the dominant broadening resulting from isotopic randomness in natural Si reveals new fine structure in the boron bound exciton luminescence. PMID- 11415417 TI - Dynamic light scattering at domains and nanoclusters in a relaxor ferroelectric. AB - In the probably first successful photon correlation spectroscopy experiment on a crystalline solid, inelastic light scattering of vibrating random-field-pinned domain walls has been observed at subkilohertz frequencies in the uniaxial relaxor ferroelectric Sr(0.61--x)CexBa(0.39)Nb(2)O(6). In the paraelectric relaxor phase the response of polar nanoclusters becomes overdamped and Rayleigh like. PMID- 11415418 TI - Variable range hopping and electrical conductivity along the DNA double helix. AB - We present a model to describe electrical conductivity along the DNA double helix. In this model, DNA is considered as a one-dimensional disordered system, and electrons are transported via variable range hopping between localized states. Thermal structural fluctuations in DNA further localize electronic wave functions, giving rise to a temperature-dependent localization length. The model quantitatively explains the temperature dependence of the conductivity observed in the lambda phage DNA (lambda-DNA). PMID- 11415419 TI - Dynamics of DNA-protein interaction deduced from in vitro DNA evolution. AB - We report the first experimental study of the dynamics of in vitro evolution of DNA. Starting from a random pool of DNA sequences, we used cycles of selection (binding to the lac repressor protein), amplification, and mutations to evolve to a unique DNA sequence, the lac operator. Statistical analysis of the DNA sequences obtained during the cycles of evolution shows that the DNA bases are selected at different rates. The rates of selection provide a quantitative measure of the interaction of the DNA bases with the protein during the complex formation. A model reproduces the evolution dynamics of the DNA population but cannot give the fine structure of the DNA-protein interaction. PMID- 11415420 TI - Behavioral-independent features of complex heartbeat dynamics. AB - We test whether the complexity of the cardiac interbeat interval time series is simply a consequence of the wide range of scales characterizing human behavior, especially physical activity, by analyzing data taken from healthy adult subjects under three conditions with controls: (i) a "constant routine" protocol where physical activity and postural changes are kept to a minimum, (ii) sympathetic blockade, and (iii) parasympathetic blockade. We find that when fluctuations in physical activity and other behavioral modifiers are minimized, a remarkable level of complexity of heartbeat dynamics remains, while for neuroautonomic blockade the multifractal complexity decreases. PMID- 11415421 TI - Dislocation-controlled perforated layer phase in a PEO- b-PS diblock copolymer. AB - Small angle x-ray analyses show that the shear-induced hexagonal perforated layer phase in a poly(ethylene oxide)- b-polystyrene diblock copolymer consists of trigonal (R3;m) twins and a hexagonal (P6(3)/mmc) structure, with trigonal twins being majority components. Transmission electron microscopy reveals that the hexagonal structure is generated through sequential intrinsic stacking faults on the second layer from a previous edge dislocation line, while the trigonal twins are formed by successive intrinsic stacking faults on neighboring layers due to the plastic deformation under mechanical shear. PMID- 11415422 TI - Wave front depinning transition in discrete one-dimensional reaction-diffusion systems. AB - Pinning and depinning of wave fronts are ubiquitous features of spatially discrete systems describing a host of phenomena in physics, biology, etc. A large class of discrete systems is described by overdamped chains of nonlinear oscillators with nearest-neighbor coupling and controlled by constant external forces. A theory of the depinning transition for these systems, including scaling laws and asymptotics of wave fronts, is presented and confirmed by numerical calculations. PMID- 11415423 TI - Formation of two-dimensional concentration pulses on microdesigned composite catalyst surfaces. AB - We study the effect of microdesigned composite geometries on pattern formation during the catalytic oxidation of CO on Pt-Ti, Pt-Rh, and Pt-Pd composite catalysts. In particular, we find experimentally (and rationalize through modeling) that adsorbate surface transport through the second (active) component drastically affects the shapes and interactions of concentration patterns (traveling pulses) observed on pure Pt. PMID- 11415424 TI - Glasslike kinetic arrest at the colloidal-gelation transition. AB - We show that gelation of weakly attractive colloids is remarkably similar to the colloidal glass transition. Like the glass transition, dynamic light scattering functions near gelation scale with scattering vector, and exhibits a two-step decay with a power-law divergence of the final decay time. Like the glass transition, static light scattering does not change upon gelation. These results suggest that, like the glass transition, gelation results from kinetic arrest due to crowding of clusters, and that both gelation and the glass transition are manifestations of a more general jamming transition. PMID- 11415425 TI - Comment on "Pipe network model for scaling of dynamic interfaces in porous media". PMID- 11415427 TI - Comment on "Upper limit to Landau damping in helicon discharges". PMID- 11415428 TI - Comment on "Charge-orbital stripe structure in La(1--x)Ca(x)MnO(3) (x = 1/2,2/3)". PMID- 11415429 TI - Functional role of polar amino acid residues in Na+/H+ exchangers. AB - Na(+)/H(+) exchangers are a family of ubiquitous membrane proteins. In higher eukaryotes they regulate cytosolic pH by removing an intracellular H(+) in exchange for an extracellular Na(+). In yeast and Escherichia coli, Na(+)/H(+) exchangers function in the opposite direction to remove intracellular Na(+) in exchange for extracellular H(+). Na(+)/H(+) exchangers display an internal pH sensitivity that varies with the different antiporter types. Only recently have investigations examined the amino acids involved in pH-sensitivity and in cation binding and transport. Histidine residues are good candidates for H(+)-sensing amino acids, since they can ionize within the physiological pH range. Histidine residues have been shown to be important in the function of the E. coli Na(+)/H(+) exchanger NhaA and in the yeast Na(+)/H(+) exchanger sod2. In E. coli, His(225) of NhaA may function to interact with, or regulate, the pH-sensory region of NhaA. In sod2, His(367) is also critical to transport and may be a functional analogue of His(225) of NhaA. Histidine residues are not critical for the function of the mammalian Na(+)/H(+) exchanger, although an unusual histidine rich sequence of the C-terminal tail has some influence on activity. Other amino acids involved in cation binding and transport by Na(+)/H(+) exchangers are only beginning to be studied. Amino acids with polar side chains such as aspartate and glutamate have been implicated in transport activity of NhaA and sod2, but have not been studied in the mammalian Na(+)/H(+) exchanger. Further studies are needed to elucidate the mechanisms involved in pH-sensitivity and cation binding and transport by Na(+)/H(+) exchangers. PMID- 11415430 TI - Cholesterol depletion reduces apical transport capacity in epithelial Madin-Darby canine kidney cells. AB - Reduction of the cholesterol level in membranes of epithelial Madin-Darby canine kidney (MDCK) cells reverses the apical-to-basolateral transport ratio of the apical membrane marker protein influenza virus haemagglutinin and the secreted glycoprotein gp80. At the same time, basolateral transport of the vesicular stomatitis virus G protein is unaffected [Keller and Simons (1998) J. Cell Biol. 140, 1357-1367]. To investigate whether cholesterol depletion influences apical sorting mechanisms specifically, or apical transport capacity more generally, we studied the effect of cholesterol depletion on the secretion of three different classes of molecules from the apical and basolateral surfaces of MDCK cell layers: glycoprotein gp80, sulphated proteoglycans and proteins, and non glycosylated rat growth hormone. In each case, cholesterol depletion reduced the fraction secreted to the apical medium and increased the fraction secreted basolaterally. The fact that this was observed for all sulphated proteins and proteoglycans and for the non-glycosylated rat growth hormone, which is randomly secreted in untreated cells, indicates that cholesterol depletion reduces the apical transport capacity, rather than interfering with specific recognition and sorting processes. PMID- 11415431 TI - Intracellular distribution of glycogen synthase and glycogen in primary cultured rat hepatocytes. AB - Changes in the intracellular distribution of liver glycogen synthase (GS) might constitute a new regulatory mechanism for the activity of this enzyme at cellular level. Our previous studies indicated that incubation of isolated hepatocytes with glucose activated GS and resulted in its translocation from a homogeneous cytosolic distribution to the cell periphery. These studies also suggested a relationship with insoluble elements of the cytoskeleton, in particular actin. Here we show the translocation of GS in a different experimental model that allows the analysis of this phenomenon in long-term studies. We describe the reversibility of translocation of GS and its effect on glycogen distribution. Incubation of cultured rat hepatocytes with glucose activated GS and triggered its translocation to the hepatocyte periphery. The relative amount of the enzyme concentrated near the plasma membrane increased with time up to 8 h of incubation with glucose, when the glycogen stores reached their maximal value. The lithium induced covalent activation of GS was not sufficient to cause its translocation to the cell periphery. The intracellular distribution of GS closely resembled that of glycogen. Our results showed an interaction between GS and an insoluble element of the hepatocyte matrix. Although no co-localization between actin filaments and GS was observed in any condition, disruption of actin cytoskeleton resulted in a significantly lower percentage of cells in which the enzyme translocated to the cell periphery in response to glucose. This observation suggests that the microfilament network has a role in the translocation of GS. PMID- 11415432 TI - Characterization of Sp17: a ubiquitous three domain protein that binds heparin. AB - Sp17 is a protein that was originally thought to be expressed exclusively in the testis and whose primary function was binding to the extracellular matrix of the oocyte. Several recent reports have implicated Sp17 as having a role in cell-cell adhesion and/or cell migration in transformed, lymphocytic and haematopoietic cells, possibly through its interaction with extracellular heparan sulphate. In the present study, we report that Sp17's central domain (amino acids 61-117), spanning exon 3, is critical for heparin binding. Sp17 has two additional functional domains, an N-terminal domain similar to the dimer-interaction site in the cAMP-dependent protein kinase IIalpha regulatory subunit and a C-terminal calmodulin-binding domain. The mouse gene for Sp17 is 6.5 kb and contains four exons. Although Sp17 expression is highest in the testis, it is present in all of the mouse somatic tissues examined and is highly conserved throughout all mammalian species. Sp17's central domain, which is necessary for heparin binding, exhibits the greatest sequence divergence of all three domains. The Sp17 gene is induced in metastatic cells and during mucosal immune responses, and the protein appears to play an important role in cell migration and/or adhesion in somatic cells, as well as in male germ cells. PMID- 11415433 TI - The mouse Nudt7 gene encodes a peroxisomal nudix hydrolase specific for coenzyme A and its derivatives. AB - A mouse homologue of the Saccharomyces cerevisiae Pcd1p coenzyme A diphosphatase, NUDT7alpha, has been expressed as a thioredoxin fusion protein in Escherichia coli. NUDT7alpha is also a CoA diphosphatase of the nudix hydrolase family, and hydrolyses CoA, CoA esters and oxidized CoA with similar efficiences, yielding 3',5'-ADP and the corresponding 4'-phosphopantetheine derivative as products. K(m) and k(cat) values with CoA were 240 microM and 3.8 s(-1). Activity was optimal at pH 8.0 with 5 mM Mg(2+) or Mn(2+) ions, while fluoride was inhibitory with an IC(50) value of 20 microM. Expression of the Nudt7 gene was highest in liver, intermediate in lung and kidney, and lowest in brain and heart, producing a 1.5 kb transcript. A similar pattern of expression was found for the human orthologue, NUDT7. An enzymically inactive splice variant, NUDT7beta, which lacks 20 amino acids downstream of the nudix motif, was also found to be expressed in mouse tissues. Transfection of HeLa cells with a vector expressing the Nudt7alpha gene fused to the C-terminus of red fluorescent protein showed that NUDT7alpha, like Pcd1p, was a peroxisomal enzyme. The function of the NUDT7 enzyme may be the elimination of oxidized CoA from peroxisomes, or the regulation of CoA and acyl CoA levels in this organelle in response to metabolic demand. PMID- 11415434 TI - Protein kinase C-delta C2-like domain is a binding site for actin and enables actin redistribution in neutrophils. AB - Neutrophils play a key role in host-defence mechanisms against invading pathogens, using their capacity to migrate, engulf micro-organisms and produce toxic radicals. Protein kinase C (PKC) isotypes are important intracellular regulators of these processes in neutrophils. PKC isotypes themselves are controlled by interactions with lipids, Ca(2+) and proteins. The C2-like domain of PKC-delta (deltaC2) has been identified as a protein-interaction domain in this PKC isotype. In the present paper we have investigated the contribution of protein interactions at this domain to the regulation/function of PKC-delta in neutrophils. Using affinity chromatography we identified actin as a deltaC2 binding partner in these cells. Fluorescein-labelled deltaC2, microinjected into immobilized neutrophils, interacts with filamentous actin (F-actin) inside the cell. PKC-delta co-localizes with F-actin in neutrophils, in lamellipodia at the leading edge of the cell. Stimulation with phorbol ester or IgG-opsonized Staphylococcus aureus results in co-ordinated redistribution of PKC-delta and F actin, and a PKC-delta inhibitor inhibits these changes. Microinjection of deltaC2 also inhibits F-actin redistribution. Thus PKC-delta binds to F-actin through its C2 domain, and these interactions are important in regulating actin redistribution in neutrophils. PMID- 11415435 TI - Production and characterization of recombinant human CLN2 protein for enzyme replacement therapy in late infantile neuronal ceroid lipofuscinosis. AB - Late infantile neuronal ceroid lipofuscinosis (LINCL) is a fatal recessive childhood disease caused by mutations in the CLN2 gene, which encodes the lysosomal enzyme tripeptidyl peptidase I. As a step towards understanding the protein and developing therapeutics for the disease, we have produced and characterized recombinant human CLN2 (ceroid lipofuscinosis, neuronal 2) protein from Chinese-hamster ovary cells engineered to secrete high levels of the enzyme. The protein was secreted as an inactive soluble proenzyme of approximately 65 kDa that appears as a monomer by gel filtration. Upon acidification, the protein is processed to mature form and acquires activity. The enzyme is efficiently delivered to the lysosomes of LINCL fibroblasts by mannose 6-phosphate-receptor mediated endocytosis (EC(50) approximately 2 nM), where it remains active for long periods of time (t(1/2) approximately 12 days). In addition, the enzyme is taken up by rat cerebellar granule neurons by mannose 6-phosphate-dependent and independent mechanisms. Treatment of LINCL fibroblasts with recombinant CLN2 protein restores normal enzyme activity and ameliorates accumulation of the major storage protein, mitochondrial ATP synthase subunit c. PMID- 11415436 TI - Positional importance of Pro53 adjacent to the Arg49-Gly50-Asp51 sequence of rhodostomin in binding to integrin alphaIIbbeta3. AB - Rhodostomin (RHO), a disintegrin isolated from snake venom, has been demonstrated to inhibit platelet aggregation through interaction with integrin alphaIIbbeta3, but there is a lack of direct evidence for RHO-integrin alphaIIbbeta3 binding. In addition, no study on the length of Arg(49)-Gly(50)-Asp(51) (RGD) loop of RHO influencing on its binding to integrin alphaIIbbeta3 has been reported. In the present study we have developed a highly sensitive dot-blot and glutathione S transferase-RHO pull-down assays; the latter was coupled with a biotin-avidin horseradish peroxidase enhanced-chemiluminescence detection system. These were able to demonstrate the direct binding of RHO to integrin alphaIIbbeta3. The pull down assay further showed that four alanine-insertion mutants upstream of the RGD motif and three insertions downstream of the RGD were able to decrease integrin alphaIIbbeta3 binding activity to only a limited extent. By contrast, two insertions immediately next to RGD and one insertion in front of the Cys(57) caused almost complete loss of binding activity to alphaIIbbeta3. The results of the platelet-aggregation-inhibition assay and platelet-adhesion assay for the insertion mutants were consistent with results of the pull-down assay. It is thus concluded that, although an insertion of a single alanine residue in many positions of the RGD loop has only minor effects on RHO binding to integrin alphaIIbbeta3, the specific position of Pro(53) residue adjacent to the RGD sequence is important for RHO binding to platelet integrin alphaIIbbeta3. PMID- 11415437 TI - Glutathione S-transferases as antioxidant defence agents confer pyrethroid resistance in Nilaparvata lugens. AB - Selection of a laboratory colony of the brown planthopper Nilaparvata lugens with the pyrethroids permethrin and lambda-cyhalothrin increased its resistance to both insecticides. Biochemical analysis and synergistic studies with metabolic inhibitors indicated that elevated glutathione S-transferases (GSTs) with a predominant peroxidase activity conferred resistance to both pyrethroids, whereas esterases conferred part of the resistance to permethrin. Purified esterases hydrolysed permethrin at a slow rate, but incubation of either pyrethroid or their primary metabolites with partially purified GSTs had no effect on the metabolic profile. Although GSTs were sensitive to inhibition by both pyrethroids, they did not serve as binding proteins, as previously hypothesized [Grant and Matsumura (1988) Insect Biochem. 18, 615-622]. We demonstrate that pyrethroids, in addition to their neurotoxic effect, induce oxidative stress and lipid peroxidation in insects. Pyrethroid exposure induced lipid peroxides, protein oxidation and depleted reduced glutathione. Elevated GSTs in the resistant strains attenuated the pyrethroid-induced lipid peroxidation and reduced mortality, whereas their in vivo inhibition eliminated their protective role. We therefore hypothesize that the main role of elevated GSTs in conferring resistance in N. lugens is through protecting tissues from oxidative damage. Our study extends the GSTs' range of efficacy to pyrethroid insecticides and possibly explains the role of elevated GSTs in other pyrethroid-resistant insects. PMID- 11415438 TI - Identification of the glycosylation sites utilized on the V1a vasopressin receptor and assessment of their role in receptor signalling and expression. AB - Most of the large family of G-protein-coupled receptors (GPCRs) possess putative N-glycosylation sites within their N-termini. However, for the vast majority of GPCRs, it has not been determined which, if any, of the consensus glycosylation sites are actually utilized or what the functional ramifications are of modification by oligosaccharide. The occurrence and function of glycosylation of the V(1a) vasopressin receptor (V(1a)R) has been investigated in this study. Using a combination of translation systems that are either glycosylation competent or do not support glycosylation, we established that of the four putative N-glycosylation sites at Asn(14), Asn(27), Asn(198) and Asn(333) only the first three sites are actually modified by carbohydrate. This was confirmed by disruption of consensus sites by site-directed mutagenesis, individually and in combination. The V(1a)R is not O-glycosylated. The functionality of a series of glycosylation-defective V(1a)R constructs was characterized after expression in HEK 293T cells. It was found that carbohydrate moieties are not required for the receptor to bind any of the four classes of ligand available, or for intracellular signalling. The glycosylation status of the V(1a)R did, however, regulate the level of total receptor expression and also the abundance of receptor at the cell surface. Furthermore, the nature of this regulation (increased or decreased expression) was dictated by the locus of the oligosaccharide modification. Modification of any one of the consensus sites alone, however, was sufficient for wild-type expression, indicating a redundancy within the glycosylation sites. A role for the carbohydrate in the correct folding or stabilization of the V(1a)R is indicated. Glycosylation is not required, however, for efficient trafficking of the receptor to the cell surface. This study establishes the functional importance of N-glycosylation of the V(1a)R. PMID- 11415439 TI - Functional roles and efficiencies of the thioredoxin boxes of calcium-binding proteins 1 and 2 in protein folding. AB - The rat luminal endoplasmic-recticulum calcium-binding proteins 1 and 2 (CaBP1 and CaBP2 respectively) are members of the protein disulphide-isomerase (PDI) family. They contain two and three thioredoxin boxes (Cys-Gly-His-Cys) respectively and, like PDI, may be involved in the folding of nascent proteins. We demonstrate here that CaBP1, similar to PDI and CaBP2, can complement the lethal phenotype of the disrupted Saccharomyces cerevisiae PDI gene, provided that the natural C-terminal Lys-Asp-Glu-Leu sequence is replaced by His-Asp-Glu Leu. Both the in vitro RNase AIII-re-activation assays and in vivo pro (carboxypeptidase Y) processing assays using CaBP1 and CaBP2 thioredoxin (trx) box mutants revealed that, whereas the three trx boxes in CaBP2 seem to be functionally equivalent, the first trx box of CaBP1 is significantly more active than the second trx box. Furthermore, only about 65% re-activation of denatured reduced RNase AIII could be obtained with CaBP1 or CaBP2 compared with PDI, and the yield of PDI-catalysed reactions was significantly reduced in the presence of either CaBP1 or CaBP2. In contrast with PDI, neither CaBP1 nor CaBP2 could catalyse the renaturation of denatured glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which is a redox-independent process, and neither protein had any effect on the PDI-catalysed refolding of GAPDH. Furthermore, although PDI can bind peptides via its b' domain, a property it shares with PDIp, the pancreas-specific PDI homologue, and although PDI can bind malfolded proteins such as 'scrambled' ribonuclease, no such interactions could be detected for CaBP2. We conclude that: (1) both CaBP2 and CaBP1 lack peptide-binding activity for GAPDH attributed to the C-terminal region of the a' domain of PDI; (2) CaBP2 lacks the general peptide-binding activity attributed to the b' domain of PDI; (3) interaction of CaBP2 with substrate (RNase AIII) is different from that of PDI and substrate; and (4) both CaBP2 and CaBP1 may promote oxidative folding by different kinetic pathways. PMID- 11415440 TI - Transcriptional regulation of the human cystathionine beta-synthase -1b basal promoter: synergistic transactivation by transcription factors NF-Y and Sp1/Sp3. AB - Cystathionine beta-synthase (CBS) catalyses the condensation of serine and homocysteine to form cystathionine, an intermediate step in the synthesis of cysteine. Human CBS encodes five distinct 5' non-coding exons, the most frequent termed CBS -1a and CBS -1b, each transcribed from its own unique GC-rich TATA less promoter. The minimal transcriptional region (-3792 to -3667) of the CBS -1b promoter was defined by 5'- and 3'-deletions, and transient transfections of reporter gene constructs in HepG2 cells, characterized by CBS transcription exclusively from the -1b promoter. Included in this 125 bp region are 3 GC-boxes (termed GC-a, GC-b and GC-c), an inverted CAAT-box and an E-box. By gel-shift and supershift assays, binding of specificity protein (Sp)1 and Sp3 to the GC-box elements, upstream stimulatory factor 1 (USF-1) to the E-box, and both nuclear factor (NF)-Y and an NF-1-like factor to the CAAT box could be demonstrated. By transient trans fections and reporter gene assays in HepG2 and Drosophila SL2 cells, a functional interplay was indicated between NF-Y binding to the CAAT-box, or between USF-1 binding to the E-box, and Sp1/Sp3 binding to the GC-box elements. In SL2 cells, NF-Y and Sp1/Sp3 were synergistic. Furthermore, both Sp1 and the long Sp3 isoform transactivated the CBS -1b minimal promoter; however, the short Sp3 isoforms were potent repressors. These results may explain the cell or tissue-specific regulation of CBS transcription, and clarify the bases for alterations in CBS gene expression in human disease and Down's syndrome. PMID- 11415441 TI - Activation of pro-(matrix metalloproteinase-2) (pro-MMP-2) by thrombin is membrane-type-MMP-dependent in human umbilical vein endothelial cells and generates a distinct 63 kDa active species. AB - Thrombin, a critical enzyme in the coagulation cascade, has also been associated with angiogenesis and activation of the zymogen form of matrix metalloproteinase 2 (MMP-2 or gelatinase-A). We show that thrombin activated pro-MMP-2 in a dose- and time-dependent manner in cultured human umbilical-vein endothelial cells (HUVECs) to generate a catalytically active 63 kDa protein that accumulated as the predominant form in the conditioned medium. This 63 kDa thrombin-activated MMP-2 is distinct from the 62 kDa species found following concanavalin A or PMA stimulated pro-MMP-2 activation. Hirudin and leupeptin blocked thrombin-induced pro-MMP-2 activation, demonstrating that the proteolytic activity of thrombin is essential. However, activation was also dependent upon membrane-type-MMP (MT-MMP) action, since it was blocked by EDTA, o-phenanthroline, hydroxamate metalloproteinase inhibitors, tissue inhibitor of metalloproteinase-2 (TIMP-2) and TIMP-4, but not TIMP-1. Thrombin inefficiently cleaved recombinant 72 kDa pro MMP-2, but efficiently cleaved the 64 kDa MT-MMP-processed intermediate form in the presence of cells. Thrombin also rapidly (within 1 h) increased cellular MT MMP activity, and at longer time points (>6 h) it increased expression of MT1-MMP mRNA and protein. Thus signalling via proteinase-activated receptors (PARs) may play a role in thrombin-induced MMP-2 activation, though this does not appear to involve PAR1, PAR2, or PAR4 in HUVECs. These results indicate that in HUVECs the activation of pro-MMP-2 by thrombin involves increased MT-MMP activity and preferential cleavage of the MT-MMP-processed 64 kDa MMP-2 form in the presence of cells. The integration of these proteinase systems in the vascular endothelium may be important during thrombogenesis and tissue remodelling associated with neovascularization. PMID- 11415442 TI - Roles of triosephosphate isomerase and aerobic metabolism in Trypanosoma brucei. AB - Kinetoplastid protozoa compartmentalize the first seven enzymes of glycolysis and two enzymes of glycerol metabolism in a microbody, the glycosome. While in its mammalian host, Trypanosoma brucei depends entirely on glucose for ATP generation. Under aerobic conditions, most of the glucose is metabolized to pyruvate. Aerobic metabolism depends on the activities of glycosomal triosephosphate isomerase and a mitochondrial glycerophosphate oxidase, and on glycerophosphate<-->dihydroxyacetone phosphate exchange across the glycosomal membrane. Using a combination of genetics and computer modelling, we show that triosephosphate isomerase is probably essential for bloodstream trypanosome survival, but not for the insect-dwelling procyclics, which preferentially use amino acids as an energy source. When the enzyme level decreased to about 15% of that of the wild-type, the growth rate was halved. Below this level, a lethal rise in dihydroxyacetone phosphate was predicted. Expression of cytosolic triosephosphate isomerase inhibited cell growth. Attempts to knockout the trypanosome alternative oxidase genes (which are needed for glycerophosphate oxidase activity) were unsuccessful, but when we lowered the level of the corresponding mRNA by expressing a homologous double-stranded RNA, oxygen consumption was reduced fourfold and the rate of trypanosome growth was halved. PMID- 11415443 TI - Molecular cloning and characterization of chemokine-like factor 1 (CKLF1), a novel human cytokine with unique structure and potential chemotactic activity. AB - Cytokines are small proteins that have an essential role in the immune and inflammatory responses. The repertoire of cytokines is becoming diverse and expanding. Here we report the identification and characterization of a novel cytokine designated as chemokine-like factor 1 (CKLF1). The full-length cDNA of CKLF1 is 530 bp long and a single open reading frame encoding 99 amino acid residues. CKLF1 bears no significant similarity to any other known cytokine in its amino acid sequence. Expression of CKLF1 can be partly inhibited by interleukin 10 in PHA-stimulated U937 cells. Recombinant CKLF1 is a potent chemoattractant for neutrophils, monocytes and lymphocytes; moreover, it can stimulate the proliferation of murine skeletal muscle cells. These results suggest that CKLF1 might have important roles in inflammation and in the regeneration of skeletal muscle. PMID- 11415444 TI - Elevated extracellular [K+] inhibits death-receptor- and chemical-mediated apoptosis prior to caspase activation and cytochrome c release. AB - Efflux of intracellular K(+) and cell shrinkage are features of apoptosis in many experimental systems, and a regulatory role has been proposed for cytoplasmic [K(+)] in initiating apoptosis. We have investigated this in both death-receptor mediated and chemical-induced apoptosis. Using Jurkat T cells pre-loaded with the K(+) ion surrogate (86)Rb(+), we have demonstrated an efflux of intracellular K(+) during apoptosis that was concomitant with, but did not precede, other apoptotic changes, including phosphatidylserine externalization, mitochondrial depolarization and cell shrinkage. To further clarify the role of K(+) ions in apoptosis, cytoprotection by elevated extracellular [K(+)] was studied. Induction of apoptosis by diverse death-receptor and chemical stimuli in two cell lines was inhibited prior to phosphatidylserine externalization, mitochondrial depolarization, cytochrome c release and caspase activation. Using a cell-free system, we have demonstrated a novel mechanism by which increasing [K(+)] inhibited caspase activation. In control dATP-activated lysates, Apaf-1 oligomerized to a biologically active caspase processing approximately 700 kDa complex and an inactive approximately 1.4 MDa complex. Increasing [K(+)] inhibited caspase activation by preventing formation of the approximately 700 kDa complex, but not of the inactive complex. Thus intracellular and extracellular [K(+)] markedly affect caspase activation and the initiation of apoptosis induced by both death-receptor ligation and chemical stress. PMID- 11415445 TI - Inhibition of HIV-1 gene expression by novel macrophage-tropic DNA enzymes targeted to cleave HIV-1 TAT/Rev RNA. AB - Many regions of the HIV-1 genome have been targeted in earlier studies by RNA cleaving DNA enzymes possessing the 10-23 catalytic motif, and efficient inhibition of HIV-1 gene expression was reported. All these studies employed charged synthetic lipids to introduce the catalytic DNA into the mammalian cells, which severely limits its practical application and usefulness in vivo. Taking advantage of the ability of G residues to interact directly with the scavenger receptors on the macrophages, we synthesized a DNA enzyme 5970 that contained 10 G residues at the 3' end. With the aim of improving the intracellular stability of the DNA enzyme 5970, we added two short stretches of stem-loop structures that were 12 bases long on either side of the DNA enzyme 5970. DNA enzyme 5970 without the poly-G tracts cleaved the synthetic RNA of HIV-1 TAT/Rev, two important regulatory proteins of HIV, very efficiently in a sequence-specific manner. Addition of 10 G residues at the 3' end of the DNA enzyme affected the cleavage efficiency only marginally whereas the same DNA enzyme with stem-loop structures on either end was significantly less efficient. The DNA enzyme with the poly-G tract at its 3' end was taken up specifically by a human macrophage-specific cell line directly in the absence of Lipofectin and was also able to inhibit HIV-1 gene expression in a transient-expression system as well as when challenged with the virus. The potential applications of these novel macrophage-tropic DNA enzymes are discussed. PMID- 11415446 TI - Hsp70 regulates the interaction between the peroxisome targeting signal type 1 (PTS1)-receptor Pex5p and PTS1. AB - The peroxisome targeting signal type 1 (PTS1) receptor, Pex5p, of the tetratricopeptide repeat (TPR) motif family is located mostly in the cytosol and mediates the translocation of PTS1 proteins to peroxisomes. As a step towards understanding the mechanisms of protein import into peroxisomes, we investigated the molecular mechanisms involved in PTS1 recognition by Pex5p with regard to requirement of energy and cytosolic factors, using cell-free synthesized acyl-CoA oxidase (AOx) as a PTS1 cargo protein, together with Pex5p and heat-shock protein (Hsp)70 from rat liver. Pex5p was partly associated with peroxisomes of rat liver, was resistant to washing with a high concentration of salt and to alkaline extraction and was inaccessible to protease added externally. Pex5p bound to AOx in an ATP-dependent manner. AOx synthesized in a cell-free translating system from rabbit reticulocyte lysate was imported into peroxisomes without being supplemented with Pex5p and Hsp70, implying that peroxisome-associated Pex5p was released from the membranes and functional in this in vitro import assay. Antibodies against Pex5p and Hsp70 inhibited AOx import. In contrast, AOx synthesized in a wheat-germ lysate required the external addition of Pex5p for import, in which Hsp70 augmented the AOx import. The TPR domain of Pex5p was revealed to bind to the N-terminal part in an Hsp70-independent manner, whereas mutual interaction of the TPR region was noted in the presence of Hsp70. Hsp70 interacted with the TPR domain of Pex5p. Moreover, Hsp70 and ATP synergistically enhanced the binding of Pex5p to the C-terminal PTS1-containing part of AOx, implying that Pex5p recognizes its cargo PTS1 protein by chaperone-assisted as well as energy-dependent mechanisms in vivo. PMID- 11415447 TI - The nematode Caenorhabditis elegans synthesizes unusual O-linked glycans: identification of glucose-substituted mucin-type O-glycans and short chondroitin like oligosaccharides. AB - The free-living nematode Caenorhabditis elegans is a relevant model for studies on the role of glycoconjugates during development of multicellular organisms. Several genes coding for glycosyltransferases involved in the synthesis of N- and O-linked glycans have already been isolated, but, apart from repetitive dimers of glycosaminoglycans, no detailed structure of either type of component has been published so far. This study aimed to establish the structures of the major O glycans synthesized by C. elegans to give an insight into the endogenous glycosyltransferase activities expressed in this organism. By the use of NMR and MS, we have resolved the sequence of seven of these components that present very unusual features. Most of them were characterized by the type-1 core substituted on Gal and/or GalNAc by (beta1-4)Glc and (beta1-6)Glc residues. Another compound exhibited the GalNAc(beta1-4)N-acetylglucosaminitol sequence in the terminal position, to which was attached a tetramer of beta-Gal substituted by both Fuc and 2-O-methyl-fucose residues. Our experimental procedure led also to the isolation of glycosaminoglycan-like components and oligomannosyl-type N-glycans. In particular, the data confirmed that C. elegans synthesizes the ubiquitous linker sequence GlcA(beta1-3)Gal(beta1-3)Gal(beta1-4)Xyl. PMID- 11415448 TI - Isolation and characterization of the human stearoyl-CoA desaturase gene promoter: requirement of a conserved CCAAT cis-element. AB - Stearoyl-CoA desaturase is the rate-limiting enzyme in the production of mono unsaturated fatty acids. We have recently cloned and characterized the human Scd cDNA and SCD (the stearoyl-CoA desaturase structural gene) on chromosome 10, as well as the non-transcribed pseudogene on chromosome 17. In order to further define SCD regulation and function, we have isolated and characterized the promoter of the structural gene. Screening of chromosome-10-specific libraries resulted in the isolation of 4.1 kb of SCD sequence upstream of the translation start site. Binding sites for transcription factors critical for mouse Scd1 and Scd2 promoter activity, such as sterol-regulated-element-binding protein and nuclear factor Y, were present in the human SCD promoter (Scd is the mouse stearoyl-CoA desaturase gene). Deletion analysis in HaCaT keratinocytes identified a critical region for promoter activity between nts 496-609 upstream of the translation start site. Site-directed mutagenesis of binding sites in this region identified the CCAAT box as the critical cis-element for SCD promoter activity. An electrophoretic mobility-shift assay confirmed that this element binds nuclear proteins from HaCaT keratinocytes. The polyunsaturated-fatty-acid (PUFA) response element, previously identified in the promoters of mouse Scd1 and Scd2, was found to be conserved in the human SCD promoter, and contained the critical CCAAT cis-element. A minimal promoter construct including this region was responsive to fatty acids, with oleate and linoleate decreasing transcription and stearate increasing it. These studies indicate that CCAAT-box-binding proteins activate SCD transcription in cultured keratinocytes and that fatty acids modulate transcription, most likely through the conserved PUFA response element. PMID- 11415449 TI - Presteady-state kinetics of Bacillus 1,3-1,4-beta-glucanase: binding and hydrolysis of a 4-methylumbelliferyl trisaccharide substrate. AB - In the present study the first stopped-flow experiments performed on Bacillus 1,3 1,4-beta-glucanases are reported. The presteady-state kinetics of the binding of 4-methylumbelliferyl 3-O-beta-cellobiosyl-beta-D-glucoside to the inactive mutant E134A, and the wild-type-catalysed hydrolysis of the same substrate, were studied by measuring changes in the fluorescence of bound substrate or 4 methylumbelliferone produced. The presteady-state traces all showed an initial lag phase followed by a fast monoexponential phase leading to equilibration (for binding to E134A) or to steady state product formation (for the wild-type reaction). The lag phase, with a rate constant of the order of 100 s(-1), was independent of the substrate concentration; apparently an induced-fit mechanism governs the formation of enzyme-substrate complexes. The concentration dependencies of the observed rate constant of the second presteady-state phase were analysed according to a number of reaction models. For the reaction of the wild-type enzyme, it is shown that the fast product formation observed before steady state is not due to a rate-determining deglycosylation step. A model that can explain the observed results involves, in addition to the induced fit, a conformational change of the productive ES complex into a form that binds a second substrate molecule in a non-productive mode. PMID- 11415450 TI - Role of the cystine-knot motif at the C-terminus of rat mucin protein Muc2 in dimer formation and secretion. AB - DNA constructs based on the 534-amino-acid C-terminus of rat mucin protein Muc2 (RMC), were transfected into COS cells and the resultant (35)S-labelled dimers and monomers were detected by SDS/PAGE of immunoprecipitates. The cystine-knot construct, encoding the C-terminal 115 amino acids, appeared in cell lysates as a 45 kDa dimer, but was not secreted. A construct, devoid of the cystine knot, failed to form dimers. Site-specific mutagenesis within the cystine knot was performed on a conserved unpaired cysteine (designated Cys-X), which has been implicated in some cystine-knot-containing growth factors as being important for intermolecular disulphide-bond formation. Dimerization of RMC was effectively abolished. Each cysteine (Cys-1-Cys-6) comprising the three intramolecular disulphide bonds of the cystine knot was then mutated. Dimer formation was impaired in each case, although much less so for the Cys-3 mutant than the others. Abnormal high-molecular-mass, disulphide-dependent aggregates formed with mutations Cys-1, Cys-2, Cys-4 and Cys-5(,) and were poorly secreted. It is concluded that the intact cystine-knot domain is essential for dimerization of the C-terminal domain of rat Muc2, and that residue Cys-X in the knot plays a key role. The structural integrity of the cystine knot, maintained by intramolecular bonds Cys-1-Cys-4, Cys-2-Cys-5 and Cys-3-Cys-6, also appears to be important for dimerization, probably by allowing correct positioning of the unpaired Cys-X residue for stable intermolecular cystine-bond formation. PMID- 11415451 TI - Characterization of the cyanobacterial ycf37: mutation decreases the photosystem I content. AB - We have constructed and analysed a cyanobacterial mutant that lacks the putative homologue of ycf37, the chloroplast open reading frame 37, which is conserved in different algae, but missing in the plastome of higher plants. In this report we show that Ycf37 of Synechocystis sp. PCC 6803 contains three tetratrico-peptide repeat (TPR) units resembling the structural organization of Ycf3, a protein that has been suggested to function as a chaperone during photosystem (PS) I complex formation. We demonstrate a light-activated transcript accumulation of this gene. Inactivation of ycf37 leads to a lower PSI/PSII ratio and a higher phycocyanin/chlorophyll ratio in Synechocystis cells. The observed alterations in the ycf37 mutants and the structural organization of the gene product suggest a functional role in PSI stability or assembly. PMID- 11415452 TI - Structure-function analysis of purified Enterococcus hirae CopB copper ATPase: effect of Menkes/Wilson disease mutation homologues. AB - The Enterococcus hirae CopB ATPase (EC 3.6.1.3) confers copper resistance to the organism by expelling excess copper. Two related human ATPase genes, ATP7A (EC 3.6.1.36) and ATP7B (EC 3.6.1.36), have been cloned as the loci of mutations causing Menkes and Wilson diseases, diseases of copper metabolism. Many mutations in these genes have been identified in patients. Since it has not yet been possible to purify the human copper ATPases, it has proved difficult to test the impact of mutations on ATPase function. Some mutations occur in highly conserved sequence motifs, suggesting that their effect on function can be tested with a homologous enzyme. Here, we used the E. hirae CopB ATPase to investigate the impact of such mutations on enzyme function in vivo and in vitro. The Menkes disease mutation of Cys-1000-->Arg, changing the conserved Cys-Pro-Cys ('CPC') motif, was mimicked in CopB. The corresponding Cys-396-->Ser CopB ATPase was unable to restore copper resistance in a CopB knock-out mutant in vivo. The purified mutant ATPase still formed an acylphosphate intermediate, but possessed no detectable ATP hydrolytic activity. The most frequent Wilson disease mutation, His-1069-->Gln, was introduced into CopB as His-480-->Gln (H480Q). This mutant CopB also failed to confer copper resistance to a CopB knock-out strain. Purified H480Q CopB formed an acylphosphate intermediate and retained a small, but significant, ATPase activity. Our results reveal that Cys-396 and His-480 of CopB are key residues for ATPase function, and similar roles are suggested for Cys 1000 and His-1069 of Menkes and Wilson ATPases respectively. PMID- 11415453 TI - Active-site mutations impairing the catalytic function of the catalytic subunit of human protein phosphatase 2A permit baculovirus-mediated overexpression in insect cells. AB - Members of the phosphoprotein phosphatase (PPP) family of protein serine/threonine phosphatases, including protein phosphatase (PP)1, PP2A and PP2B, share invariant active-site residues that are critical for catalytic function [Zhuo, Clemens, Stone and Dixon (1994) J. Biol. Chem. 269, 26234-26238]. Mutation of the active-site residues Asp(88) or His(118) within the human PP2A catalytic subunit (PP2Ac)alpha impaired catalytic activity in vitro; the D88N and H118N substitutions caused a 9- and 23-fold reduction in specific activity respectively, when compared with wild-type recombinant PP2Ac, indicating an important role for these residues in catalysis. Consistent with this, the D88N and H118N substituted forms failed to provide PP2A function in vivo, because, unlike wild-type human PP2Acalpha, neither substituted for the endogenous PP2Ac enzyme of budding yeast. Relative to wild-type PP2Ac, the active-site mutants were dramatically overexpressed in High Five insect cells using the baculovirus system. Milligram quantities of PP2Ac were purified from 1x10(9) High Five cells and the kinetic constants for dephosphorylation of the peptide RRA(pT)VA (single letter amino-acid notation) by PP2Ac (K(m)=337.5 microM; k(cat)=170 s(-1)) and D88N (K(m)=58.4 microM; k(cat)=2 s(-1)) were determined. The results show that the substitution impairs catalysis severely without a significant effect on substrate binding, consistent with the PPP catalytic mechanism. Combination of the baculovirus and yeast systems provides a strategy whereby the structure function of PP2Ac may be fully explored, a goal which has previously proven difficult, owing to the stringent auto-regulatory control of PP2Ac protein levels in vivo. PMID- 11415454 TI - Homocysteine stimulates the expression of monocyte chemoattractant protein-1 receptor (CCR2) in human monocytes: possible involvement of oxygen free radicals. AB - Homocysteinaemia is an independent risk factor for atherosclerosis. The development of atherosclerosis involves monocyte chemoattractant protein 1 (MCP 1)-mediated monocyte recruitment to the lesion site. The action of MCP-1 is mostly via its interaction with MCP-1 receptor (CCR2), which is the major receptor for MCP-1 on the surface of monocytes. The objective of the present study was to investigate the effect of homocysteine on CCR2 expression in human THP-1 monocytes. Cells were incubated with various concentrations of homocysteine for 6, 12, 24 and 48 h. The expression of CCR2 mRNA was determined by nuclease protection assay and the CCR2 protein was measured by Western immunoblotting analysis. The binding of MCP-1 to CCR2 as a functional receptor on the monocyte surface was determined by flow cytometry. Homocysteine (0.05-0.2 mM) significantly enhanced the expression of CCR2 mRNA (129-209% of the control) and CCR2 protein (up to 183% of control) in these cells after 24 h of incubation. Stimulation of CCR2 expression was associated with a parallel increase in the binding activity of CCR2 (129-191% of control) as well as an enhanced chemotactic response of homocysteine-treated monocytes. Further investigation revealed that the levels of superoxide were significantly elevated in cells incubated with homocysteine for 12-48 h. The addition of superoxide dismutase, a scavenger of superoxide, to the culture medium abolished the stimulatory effect of homocysteine on CCR2 expression as well as the binding activity of the receptor. The stimulatory effect of homocysteine on the expression of CCR2 mRNA and the levels of CCR2 protein was also observed in human peripheral blood monocytes. In conclusion, the present study has clearly demonstrated that homocysteine stimulates CCR2 expression in monocytes, leading to an enhanced binding activity and chemotatic response. Homocysteine-induced superoxide formation might serve as one of the underlying mechanisms for this effect. PMID- 11415455 TI - Ferritin and the response to oxidative stress. AB - Iron is required for normal cell growth and proliferation. However, excess iron is potentially harmful, as it can catalyse the formation of toxic reactive oxygen species (ROS) via Fenton chemistry. For this reason, cells have evolved highly regulated mechanisms for controlling intracellular iron levels. Chief among these is the sequestration of iron in ferritin. Ferritin is a 24 subunit protein composed of two subunit types, termed H and L. The ferritin H subunit has a potent ferroxidase activity that catalyses the oxidation of ferrous iron, whereas ferritin L plays a role in iron nucleation and protein stability. In the present study we report that increased synthesis of both subunits of ferritin occurs in HeLa cells exposed to oxidative stress. An increase in the activity of iron responsive element binding proteins in response to oxidative stress was also observed. However, this activation was transient, allowing ferritin protein induction to subsequently proceed. To assess whether ferritin induction reduced the accumulation of ROS, and to test the relative contribution of ferritin H and L subunits in this process, we prepared stable transfectants that overexpressed either ferritin H or ferritin L cDNA under control of a tetracycline-responsive promoter. We observed that overexpression of either ferritin H or ferritin L reduced the accumulation of ROS in response to oxidant challenge. PMID- 11415456 TI - Transforming growth factor beta-1 enhances Smad transcriptional activity through activation of p8 gene expression. AB - We report that exposure of mouse embryonic fibroblasts to transforming growth factor beta-1 (TGFbeta-1) (5 ng/ml) results in a strong activation of p8 mRNA expression that precedes the induction of cell growth. Involvement of the p8 promoter in the regulation was demonstrated by using a p8-chloramphenicol acetyltransferase construct. We therefore speculated that p8 might be a mediator of TGFbeta-1 in these cells. The incorporation of [(3)H]thymidine on treatment with TGFbeta-1 was indeed significantly higher in p8(+/+) fibroblasts than in p8( /-) fibroblasts. Smad transcriptional activity was used as marker of the TGFbeta 1 signalling pathway, to probe the lower p8(-/-) response to TGFbeta-1. Two Smad binding elements (SBEs)-luciferase constructs were transfected into p8(-/-) and p8(+/+) embryonic fibroblasts before treatment with TGFbeta-1. A lower level of Smad transactivation was observed in p8(-/-) embryonic fibroblasts, under basal conditions and after stimulation with TGFbeta-1. To test whether Smad underexpression in p8(-/-) cells was actually due to p8 depletion, p8(-/-) embryonic fibroblasts were transfected with a human p8 expression plasmid together with an SBE-luciferase construct. The expression of p8 restored Smad transactivation in unstimulated and TGFbeta-1-treated cells to the level found in p8(+/+) cells. We concluded that TGFbeta-1 activates p8 expression, which in turn enhances the Smad-transactivating function responsible for TGFbeta-1 activity. PMID- 11415457 TI - Glucose regulates protein catabolism in ras-transformed fibroblasts through a lysosomal-dependent proteolytic pathway. AB - Transformed cells are exposed to heterogeneous microenvironments, including low D glucose (Glc) concentrations inside tumours. The regulation of protein turnover is commonly impaired in many types of transformed cells, but the role of Glc in this regulation is unknown. In the present study we demonstrate that Glc controls protein turnover in ras-transformed fibroblasts (KBALB). The regulation by Glc of protein breakdown was correlated with modifications in the levels of lysosomal cathepsins B, L and D, while autophagic sequestration and non-lysosomal proteolytic systems (m- and mu-calpains and the zeta-subunit of the proteasome) remained unaffected. Lactacystin, a selective inhibitor of the proteasome, depressed proteolysis, but did not prevent its regulation by Glc. The sole inhibition of the cysteine endopeptidases (cathepsins B and L, and calpains) by E 64d [(2S,3S)-trans-epoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester] was also not sufficient to alter the effect of Glc on proteolysis. The Glc-dependent increase in proteolysis was, however, prevented after optimal inhibition of lysosomal cysteine and aspartic endopeptidases by methylamine. We conclude that, in transformed cells, Glc plays a critical role in the regulation of protein turnover and that the lysosomal proteolytic capacity is mainly responsible for the control of intracellular proteolysis by Glc. PMID- 11415458 TI - Relationships between inhibition constants, inhibitor concentrations for 50% inhibition and types of inhibition: new ways of analysing data. AB - The concentration of an inhibitor that decreases the rate of an enzyme-catalysed reaction by 50%, symbolized i(0.5), is often used in pharmacological studies to characterize inhibitors. It can be estimated from the common inhibition plots used in biochemistry by means of the fact that the extrapolated inhibitor concentration at which the rate becomes infinite is equal to -i(0.5). This method is, in principle, more accurate than comparing the rates at various different inhibitor concentrations, and inferring the value of i(0.5) by interpolation. Its reciprocal, 1/i(0.5), is linearly dependent on v(0)/V, the uninhibited rate divided by the limiting rate, and the extrapolated value of v(0)/V at which 1/i(0.5) is zero allows the type of inhibition to be characterized: this value is 1 if the inhibition is strictly competitive; greater than 1 if the inhibition is mixed with a predominantly competitive component; infinite (i.e. 1/i(0.5) does not vary with v(0)/V) if the inhibition is pure non-competitive (i.e. mixed with competitive and uncompetitive components equal); negative if the inhibition is mixed with a predominantly uncompetitive component; and zero if it is strictly uncompetitive. The type of analysis proposed has been tested experimentally by examining inhibition of lactate dehydrogenase by oxalate (an uncompetitive inhibitor with respect to pyruvate) and oxamate (a competitive inhibitor with respect to pyruvate), and of cytosolic malate dehydrogenase by hydroxymalonate (a mixed inhibitor with respect to oxaloacetate). In all cases there is excellent agreement between theory and experiment. PMID- 11415459 TI - Conformational stability and warfarin-binding properties of human serum albumin studied by recombinant mutants. AB - Correctly folded recombinant wild-type human serum albumin and the single-residue mutants K199A, W214A, R218H and H242Q were produced with the use of a yeast expression system. The changes in R218H resulted in a pronounced decrease in intrinsic fluorescence. Thermodynamic parameters for thermal denaturation of the present mutants and of five additional mutants have been determined, showing small increases in stability for two mutants (R218H and H242Q) and a larger decrease in stability for one (W214A). In the last of these, denaturation was a heterogeneous process starting at physiological temperature. The high-affinity binding constant for warfarin at pH 7.4 was determined by fluorescence spectroscopy: there was a significant increase in affinity for binding of warfarin to H242Q and K199A and a smaller decrease in affinity for W214A and R218H. The findings show that Trp-214 is not as essential for the high-affinity binding of warfarin as has previously been thought. PMID- 11415460 TI - Fatty acids inhibit growth-factor-induced diacylglycerol kinase alpha activation in vascular smooth-muscle cells. AB - We have previously shown that unsaturated fatty acids amplify platelet-derived growth-factor (PDGF)-induced protein kinase C (PKC) activation in vascular smooth muscle cells (VSMCs). Diacylglycerol-induced PKC activation is normally terminated by diacylglycerol kinases (DGKs). We thus hypothesized that fatty acids act by inhibiting a DGK. Fractionation of VSMC extracts demonstrated that the DGK alpha isoform was the major DGK activity present. PDGF markedly increased the DGK activity of cultured cells. An inhibitor selective for the DGK alpha isoform, R59949 [3-[2-[4-(bis-(4-fluorophenyl)methylene]piperidin-1-yl)ethyl]-2,3 dihydro-2-thioxo-4(1H)-quinazolinone], abolished the growth-factor-induced increase in DGK activity, but had little effect on basal activity. PDGF thus selectively activates DGKalpha. Epidermal growth factor and alpha-thrombin stimulated total DGK activity similarly to PDGF. Activation by epidermal growth factor was sensitive to R59949, again suggesting involvement of DGKalpha. However, the alpha-thrombin-induced activity was unaffected by this agent. Unsaturated fatty acids inhibited growth-factor-induced DGKalpha activation, but had no effect on basal activity. Fatty acids also amplified the PDGF-induced increase in cell diacylglycerol content. These results indicate that inhibition of DGKalpha contributes to fatty-acid-induced amplification of PKC activation. Increased levels of fatty acids in diabetes may thus contribute to chronic PKC activation associated with this disorder. PMID- 11415461 TI - Characterization of the reaction mechanism for the XL-I form of bovine liver xenobiotic/medium-chain fatty acid:CoA ligase. AB - The XL-I form of xenobiotic/medium-chain fatty acid:CoA ligase was purified to apparent homogeneity from bovine liver mitochondria and used to determine the reaction mechanism. A tersubstrate kinetic analysis was conducted by varying the concentrations of ATP, benzoate and CoA in turn. Both ATP and benzoate gave parallel double-reciprocal plots against CoA, which indicates a Ping Pong mechanism, with either pyrophosphate or AMP leaving before the binding of CoA. Addition of pyrophosphate to the assays changed the plots from parallel to intersecting; addition of AMP did not. This indicates that pyrophosphate is the product that leaves before binding of CoA. Based on end-product inhibition studies, it was concluded that the reaction follows a Bi Uni Uni Bi Ping Pong mechanism, with ATP binding first, followed in order by benzoate binding, pyrophosphate release, CoA binding, benzoyl-CoA release and AMP release. A similar mechanism was obtained when the ligase was examined with butyrate as substrate. However, butyrate activation was characterized by a much higher affinity for CoA. This is attributed to steric factors resulting from the bulkier nature of the benzoate molecule. Also, with butyrate there is a bivalent cation activation distinct from that associated with binding to ATP. This activation by excess Mg(2+) results in non-linear plots of 1/v against 1/[ATP] for butyrate unless the concentrations of Mg(2+) and ATP are varied together. PMID- 11415462 TI - Analysis of a two-domain binding site for the urokinase-type plasminogen activator-plasminogen activator inhibitor-1 complex in low-density-lipoprotein receptor-related protein. AB - The low-density-lipoprotein-receptor (LDLR)-related protein (LRP) is composed of several classes of domains, including complement-type repeats (CR), which occur in clusters that contain binding sites for a multitude of different ligands. Each approximately 40-residue CR domain contains three conserved disulphide linkages and an octahedral Ca(2+) cage. LRP is a scavenging receptor for ligands from extracellular fluids, e.g. alpha(2)-macroglobulin (alpha(2)M)-proteinase complexes, lipoprotein-containing particles and serine proteinase-inhibitor complexes, like the complex between urokinase-type plasminogen activator (uPA) and the plasminogen activator inhibitor-1 (PAI-1). In the present study we analysed the interaction of the uPA-PAI-1 complex with an ensemble of fragments representing a complete overlapping set of two-domain fragments accounting for the ligand-binding cluster II (CR3-CR10) of LRP. By ligand blotting, solid-state competition analysis and surface-plasmon-resonance analysis, we demonstrate binding to multiple CR domains, but show a preferential interaction between the uPA-PAI-1 complex and a two-domain fragment comprising CR domains 5 and 6 of LRP. We demonstrate that surface-exposed aspartic acid and tryptophan residues at identical positions in the two homologous domains, CR5 and CR6 (Asp(958,CR5), Asp(999,CR6), Trp(953,CR5) and Trp(994,CR6)), are critical for the binding of the complex as well as for the binding of the receptor-associated protein (RAP) - the folding chaperone/escort protein required for transport of LRP to the cell surface. Accordingly, the present work provides (1) an identification of a preferred binding site within LRP CR cluster II; (2) evidence that the uPA-PAI-1 binding site involves residues from two adjacent protein domains; and (3) direct evidence identifying specific residues as important for the binding of uPA-PAI-1 as well as for the binding of RAP. PMID- 11415463 TI - Mitogen-activated protein kinases control p27/Kip1 expression and growth of human melanoma cells. AB - The mitogen-activated protein kinases (MAPKs) extracellular signal-regulated protein kinase (ERK)1 and ERK2, involved in regulating cell growth and differentiation, are constitutively active in A375 and WM239 human melanoma cells. Using PD098059, an inhibitor of MAPK kinase (MEK), we investigated the role of persistently activated ERK1/2 in cell growth. The inhibition of MAPK activity induced a dose-dependent growth arrest in G(0)/G(1) phase. Correspondingly, we observed the up-regulation of the cyclin-dependent kinase (Cdk) inhibitor p27/Kip1 and hypophosphorylation of the retinoblastoma protein. Further studies showed that PD098059 treatment significantly decreased Cdk2 kinase activity, most probably owing to an augmented level of p27/Kip1 associated with cyclin E-Cdk2 complexes. The accumulation of p27/Kip1 protein in A375 cells was attributed to its increased stability. Our findings suggest that constitutively active ERK1/2 kinases contribute to the growth of melanoma cells by negative regulation of the p27/Kip1 inhibitor. PMID- 11415464 TI - Haem disorder in recombinant- and reticulocyte-derived haemoglobins: evidence for stereoselective haem insertion in eukaryotes. AB - We have used NMR spectroscopy to measure haem disorder in adult human haemoglobin (HbA) obtained from mature erythrocyte cells and from yeast expressing recombinant HbA. Reticulocyte-derived HbA contained much higher levels of haem disorder (11% alpha- and 28% beta-subunit disorder) than observed for HbA from mature erythrocytes (1.5% alpha- and 8% beta-subunit disorder). Thus, unlike in vitro combination of haem and apoHb, biosynthetic haem insertion is not random with respect to orientation, but appears to show stereoselectivity. Recombinant HbA isolated from yeast showed 32% alpha- and 45% beta-subunit haem disorder. These levels relaxed to their equilibrium positions after incubating the Hb in the ferric form. Recombinant embryonic human Hbs showed less haem disorder than recombinant HbA. The levels of haem disorder in embryonic Hbs zeta(2)epsilon(2) and zeta(2)gamma(2) appear to have their equilibrium values. We propose that, in eukaryotes, in vivo haem insertion occurs via both co-translational mechanisms and insertion via semiHb-beta. PMID- 11415465 TI - Glycopeptide N-acetylgalactosaminyltransferase specificities for O-glycosylated sites on MUC5AC mucin motif peptides. AB - The recombinant proteins of the two novel UDP-N-acetylgalactosamine (GalNAc) glycopeptide:N-acetylgalactosaminyltransferases (designated gpGaNTase-T7 and gpGaNTase-T9) were assayed with O-glycosylated products obtained from the prior action of the ubiquitous transferases (GaNTase-T1 and GaNTase-T2) towards MUC5AC mucin motif peptides (GTTPSPVPTTSTTSAP and peptides with single amino acid substitutions, GTTPSAVPTTSTTSVP and GTTPSPVPTTSITSVP, that are a reflection of mucin molecule polymorphism). gpGaNTase-T9 is known to be expressed differentially and more abundantly than gpGaNTase-T7 in some tissues; the results of in vitro glycosylation also indicates a difference in acceptor substrate specificities between the gpGaNTase isoforms. With the use of capillary electrophoresis, MS and Edman degradation, our study suggests that, in the O glycosylation of mucin-type proteins, approach and recognition signalling by gpGaNTase-T7 and gpGaNTase-T9 depend largely on the peptide's primary structure (for example the presence of multiple clusters of hydroxy amino acids and the number of GalNAc residues attached to the peptide backbone). O-glycosylation in terms of sites of attachment seems to be less random than previously described and, if sequential reactions are ordered throughout the Golgi stack, the complete O-glycosylation of the mucin molecules seems to be finely tuned to respond to specific damage to, or attack on, epithelia. PMID- 11415466 TI - Asparagine synthetase expression alone is sufficient to induce l-asparaginase resistance in MOLT-4 human leukaemia cells. AB - Childhood acute lymphoblastic leukaemia (ALL) is treated by combination chemotherapy with a number of drugs, always including the enzyme L-asparaginase (ASNase). Although the initial remission rate is quite high, relapse and associated drug resistance are a significant problem. In vitro studies have demonstrated increased asparagine synthetase (AS) expression in ASNase-resistant cells, which has led to the hypothesis that elevated AS activity permits drug resistant survival. The data presented show that not only is elevated AS expression a property of ASNase-resistant MOLT-4 human leukaemia cells, but that short-term (12 h) treatment of the cells with ASNase causes a relatively rapid induction of AS expression. The results also document that the elevated expression of AS in ASNase-resistant cells is not fully reversible, even 6 weeks after ASNase removal from the culture medium. Furthermore, ASNase resistance, assessed as both drug-insensitive cell growth rates and decreased drug-induced apoptosis, parallels this irreversible AS expression. Mimicking the elevated AS activity in ASNase-resistant cells by overexpression of the human AS protein by stable retroviral transformation of parental MOLT4 cells is sufficient to induce the ASNase-resistance phenotype. These data document that ASNase resistance in ALL cells is a consequence of elevated AS expression and that although other drug induced metabolic changes occur, they are secondary to the increased asparagine biosynthetic rate. PMID- 11415468 TI - Penis size: Survey of female perceptions of sexual satisfaction. AB - BACKGROUND: Does the size of the male penis, in terms of length or width, make a difference in female sexual satisfaction? METHOD: To study the effect of penis width vs. length on female sexual satisfaction, 50 sexually active female undergraduate students were asked which felt better, i. e., was penis width or length more important for their sexual satisfaction. RESULTS: None reported they did not know, or that width and length were equally satisfying. A large majority, 45 of 50, reported width was more important (p <.001). CONCLUSION: Implications are discussed, including the fact that the data seem to contradict Masters and Johnson about penis size having no physiological effect on female sexual satisfaction. PMID- 11415467 TI - Cytosolic acidification as a signal mediating hyperosmotic stress responses in Dictyostelium discoideum. AB - BACKGROUND: Dictyostelium cells exhibit an unusual response to hyperosmolarity that is distinct from the response in other organisms investigated: instead of accumulating compatible osmolytes as it has been described for a wide range of organisms, Dictyostelium cells rearrange their cytoskeleton and thereby build up a rigid network which is believed to constitute the major osmoprotective mechanism in this organism. To gain more insight into the osmoregulation of this amoeba, we investigated physiological processes affected under hyperosmotic conditions in Dictyostelium. RESULTS: We determined pH changes in response to hyperosmotic stress using FACS or 31P-NMR. Hyperosmolarity was found to acidify the cytosol from pH 7.5 to 6.8 within 5 minutes, whereas the pH of the endo lysosomal compartment remained constant. Fluid-phase endocytosis was identified as a possible target of cytosolic acidification, as the inhibition of endocytosis observed under hypertonic conditions can be fully attributed to cytosolic acidification. In addition, a deceleration of vesicle mobility and a decrease in the NTP pool was observed. CONCLUSION: Together, these results indicate that hyperosmotic stress triggers pleiotropic effects, which are partially mediated by a pH signal and which all contribute to the downregulation of cellular activity. The comparison of our results with the effect of hyperosmolarity and intracellular acidification on receptor-mediated endocytosis in mammalian cells reveals striking similarities, suggesting the hypothesis of the same mechanism of inhibition by low internal pH. PMID- 11415469 TI - The changing distribution of two riverine tsetse flies over 15 years in an increasingly cultivated area of Burkina Faso. AB - Changes in the distribution of two riverine tsetse flies, Glossina tachinoides Westwood and Glossina palpalis gambiensis Vanderplank are described in an agro pastoral area of Burkina Faso subject to increasing human population pressure and land use change. Two similar entomological surveys (one trap every 100 m, 120 km of river) were conducted in 1981 and 1996. Changes in tsetse distribution were compared to land use changes through high resolution remote sensing imagery (LANDSAT, SPOT). There was a close relationship between proximity of crops relative to riverine forest and the density of Glossina. Where fields encroached on riverine vegetation, tsetse populations declined. Where the geomorphological structure was not well suited to agricultural activity, riverine vegetation and tsetse fly populations were relatively unaffected, even with intense agricultural activity nearby. In contrast, increased human activity and higher cattle densities in the surrounding savannah areas were associated with increased tsetse numbers. The results demonstrated a wide diversity of tsetse distribution and habitat within a few kilometres in an agro-pastoral landscape in West Africa. PMID- 11415470 TI - Effectiveness of introduced biocontrol insects on the weed Parthenium hysterophorus (Asteraceae) in Australia. AB - Six species of insects and a rust fungus have been successfully established for biocontrol of the weed Parthenium hysterophorus L. in Queensland, Australia. Effectiveness of biocontrol insects was evaluated at two properties in Queensland during 1996-97 based on an exclusion experiment using insecticides. Parthenium infested plots with and without biocontrol insects were sampled at monthly intervals and the impact of biocontrol insects on parthenium at individual plant and whole population levels monitored. Biocontrol insects were more effective at Mt Panorama (central Queensland) than at Plain Creek (north Queensland). At Mt Panorama, the leaf-feeding beetle Zygogramma bicolorata Pallister caused 96% defoliation and the stem-galling moth Epiblema strenuana Walker affected 100% of the plants, resulting in reductions of 90% in weed density, 40% in plant height, and 82% in flower production. Exclusion of biocontrol insects resulted in a 52% increase in seedling emergence and a seven-fold increase in the soil seed bank in the following season. At Plain Creek, E. strenuana was the only prominent agent. It affected 92% of the plants and prevented 32% of plants from producing any flowers, reduced plant height by 40% and flower production by 49%, but did not reduce the plant biomass, weed density or soil seed bank. However, exclusion of biocontrol insects resulted in an eight-fold increase in the soil seed bank in the following season. PMID- 11415471 TI - A density-dependent model with reinvasion for estimating tsetse fly populations (Diptera: Glossinidae) through trapping. AB - A simple density-dependent reinvasion model is described and used to estimate tsetse fly populations on the basis of removal trapping experiments. The model was tested on Glossina fuscipes fuscipes Newstead in the Central African Republic and G. palpalis palpalis (Robineau-Desvoidy) in the Republic of Congo (Brazzaville). The density-dependence is modelled by postulating that the inflow of flies each day is proportional to the deficit relative to the equilibrium population. Non-linear least square techniques were used to estimate the following parameters: the daily capture rate, the strength of the density dependence, and the equilibrium fly population, at the beginning and at the end of the trapping experiment. The model ignores birth and death rates of flies and is applicable only when a rapid decrease in population occurs over a short period (between 10 and 20 days). Over longer periods one could not ignore the natural growth of the populations as well as other more complex density-dependent mechanisms. PMID- 11415472 TI - Importance of nutrition and host availability on oogenesis and oviposition of Cephalonomia stephanoderis (Hymenoptera: Bethylidae). AB - A variety of host and non-host meals were tested for their potential in triggering oogenesis and oviposition in females of Cephalonomia stephanoderis Betrem, a synovigenic parasitoid of the coffee berry borer, Hypothenemus hampei (Ferrari). The consumption of non-host meals or meals composed of honey and crushed host eggs or pupae was insufficient to initiate oogenesis. The presence of suitable hosts for oviposition appeared essential for inducing oogenesis and oviposition in this parasitoid. Whereas females may host-feed on all developmental stages of H. hampei, these were not equally suitable in stimulating oogenesis and oviposition. In no-choice tests, oogenesis and oviposition were observed only in females supplied with fully-developed larvae, prepupae and pupae. Direct contact with hosts apparently stimulated egg maturation and egg laying. Delaying the allocation of energy to egg production until suitable hosts are available for oviposition may be an adaptive attribute of a parasitoid of a concealed and patchily distributed host such as H. hampei. PMID- 11415473 TI - Development and reproductive capacity of Thrips hawaiiensis (Thysanoptera: Thripidae) and its potential as a major pest. AB - The effect of constant temperature on development, reproduction and population growth of Thrips hawaiiensis (Morgan) reared on a diet of pollen and honey solution was studied. Although hatchability and survival rates from egg hatch to adult were higher than 79% between 15 and 25 degrees C, they were lower than 55% at 10 and 30 degrees C. Developmental rates increased linearly as rearing temperature increased. It was estimated that 153.8 degree-days, above a lower threshold temperature of 10.4 degrees C, were required to complete development from egg to adult oviposition. According to these figures, between 11 and 18 generations of T. hawaiiensis could have developed annually under outdoor conditions in western Japan, between 1990 and 1999. Mean adult longevity decreased with increasing temperature, from a maximum of 102.7 days at 15 degrees C to a minimum of 27.8 days at 25 degrees C. The mean fecundity on pollen and honey solution was highest at 20 degrees C with 536.9 eggs per female. The highest intrinsic rate of natural increase (rm) 0.208 was observed at 25 degrees C. PMID- 11415474 TI - A novel mechanism for time-sorting insect catches, and its use to derive the diel flight periodicity of brassica pod midge Dasineura brassicae (Diptera: Cecidomyiidae). PMID- 11415475 TI - Natural enemies of lepidopterous borers on maize and elephant grass in the forest zone of Cameroon. AB - The importance, geographical and temporal distributions of parasitoids of lepidopterous borers on maize and elephant grass, Pennisetum purpureum, were assessed during surveys in farmers' fields in six villages and two on-station trials in the forest zone of Cameroon between 1995 and 1996. The borer species encountered were Busseola fusca (Fuller), Sesamia calamistis Hampson, Eldana saccharina Walker on both host plants, and Mussidia nigrivenella Ragonot on maize only. Busseola fusca was the predominant host accounting for 44-57% and 96% on maize and elephant grass, respectively, followed by E. saccharina on maize with 27-39%. Fifteen hymenopterous, two dipterous and one fungal species were found on these stem and cob-borers. Among those were six pupal, six larval, four egg, one larval-pupal parasitoid and four hyperparasitoids. The scelionid parasitoids Telenomus busseolae Gahan and T. isis Polaszek were found on B. fusca eggs in all locations. During the first season, mean egg parasitism was low and ranged between 3.1% and 27% versus 54-87% during the second season. Species belonging to the Tetrastichus atriclavus Waterston complex were recovered from all four borer species. The majority and most common larval and pupal parasitoid species belonged to the ingress-and-sting guild. Larval and pupal parasitism were very erratic and on more than 50% of the sampling occasions no parasitoids were recovered. Parasitoid diversity was higher on elephant grass than maize. PMID- 11415476 TI - Spatial and temporal distribution of tsetse fly trap catches at Nguruman, southwest Kenya. AB - Spatial and temporal dynamics of rapidly growing populations of tsetse flies at Nguruman, southwest Kenya during 1993-1995, were investigated, following six years of intensive population suppression with traps over a c. 100 km2 area. The two tsetse species present were randomly distributed in the short rainy season, but were aggregated in the dry and long rainy seasons. Maximum temperature was the dominant weather factor associated with the degree of aggregation. Trends in catches at 20 fixed sites along an 18 km north-south axis were weakly correlated between locations, possibly representing population sub-structuring. In particular, trends in population change were poorly correlated between the area with a long history of trapping suppression in the south and the area with a more recent history of suppression in the north. On a micro-geographic scale, correlations among paired trap catches were clearly related to geographical proximity for Glossina pallidipes Austen (r2 = 0.55); whereas this relationship was quite weak for Glossina longipennis Corti (r2 = 0.12). Positive correlations among trap catches were significant for sites separated by less than c. 3.8 km (G. pallidipes) or 4.8 km (G. longipennis). These results suggest the existence of different population substructures in the two species on a relatively small geographic scale. PMID- 11415477 TI - Behaviour of the adult seven spot ladybird, Coccinella septempunctata (Coleoptera: Coccinellidae), in response to dimethoate residue on bean plants in the laboratory. AB - The sub-lethal effects of dimethoate residue on the distribution and locomotor activity of adults of Coccinella septempunctata Linnaeus on Vicia faba plants were investigated in the laboratory. The presence of dimethoate residues on the upper half of the plant caused C. septempunctata to spend significantly less time on the plants overall. When given a choice of treated and untreated surfaces, C. septempunctata spent proportionately more time on the untreated areas of the plant. The locomotor activity of C. septempunctata was also altered following encounter with dimethoate residues resulting in a significant increase in the proportion of time spent walking and a reduction in the proportion of time spent resting. The proportion of time spent on plant parts was also affected, with a reduction in the time spent on the apex and an increase in the proportion of time spent on the stem, indicating a possible avoidance response. The proportion of time spent on the adaxial leaf surface was significantly increased following dimethoate treatment. The implications for integrated pest management are considered. PMID- 11415478 TI - Successful in vitro rearing of Trichogramma australicum (Hymenoptera: Trichogrammatidae) on artificial diet containing cultured insect cells. PMID- 11415480 TI - Weight and body composition in a cohort of HIV-positive men and women. AB - OBJECTIVE: At issue is whether weight loss in HIV infection is a cachectic process, characterised by loss of lean body mass with conservation of fat, or a process of starvation. We present data on body composition from 516 persons at different stages of HIV infection as determined by CD4 counts. DESIGN: Cross sectional analyses of body composition in relation to CD4 count. SETTING: The baseline data from a prospective cohort study of outcomes in HIV/AIDS in relation to nutritional status in Boston, Massachusetts, USA. SUBJECTS: : The first 516 subjects with HIV/AIDS to enroll in the study. RESULTS: Differences in weight in relation to CD4 counts were present only at CD4 counts of 600 or less (slope below : 1.9 kg per 100 CD4 cells, On average, 68% of the difference in weight over CD4 counts was fat (slope: 1.3 kg fat per 100 CD4 cells, CONCLUSIONS: This cross-sectional analysis suggests that weight loss consists principally of fat loss in those persons with adequate fat stores. This observation will need to be confirmed in longitudinal analyses. PMID- 11415481 TI - Anaemia in schoolchildren in eight countries in Africa and Asia. AB - OBJECTIVE: To report on the haemoglobin concentrations and prevalence of anaemia in schoolchildren in eight countries in Africa and Asia. DESIGN: Blood samples were collected during surveys of the health of schoolchildren as a part of programmes to develop school-based health services. SETTING: Rural schools in Ghana, Indonesia, Kenya, Malawi, Mali, Mozambique, Tanzania and Vietnam. SUBJECTS: Nearly 14 000 children enrolled in basic education in three age ranges (7-11 years, 12-14 years and > or =15 years) which reflect the new UNICEF/WHO thresholds to define anaemia. RESULTS: Anaemia was found to be a severe public health problem (defined as >40% anaemic) in five African countries for children aged 7-11 years and in four of the same countries for children aged 12-14 years. Anaemia was not a public health problem in the children studied in the two Asian countries. More boys than girls were anaemic, and children who enrolled late in school were more likely to be anaemic than children who enrolled closer to the correct age. The implications of the four new thresholds defining anaemia for school-age children are examined. CONCLUSIONS: Anaemia is a significant problem in schoolchildren in sub-Saharan Africa. School-based health services which provide treatments for simple conditions that cause blood loss, such as worms, followed by multiple micronutrient supplements including iron, have the potential to provide relief from a large burden of anaemia. PMID- 11415482 TI - Seasonal aspects of weight-for-age in young children in Zimbabwe. AB - OBJECTIVE: To identify the season with the highest prevalence of underweight among young children and to examine geographical variation in seasonality of underweight. DESIGN: This analysis is based on monthly data from a clinic-based growth monitoring programme that forms part of the National Health Information System. A regression-based technique is used to identify seasonal patterns in both underweight prevalence and attendance nationally and in 60 different districts. SETTING: The analysis covers the period 1988-1995 and is based in Zimbabwe. SUBJECTS: The analysis is based on weight-for-age measurements of Zimbabwean children less than 5 years old, who attended health centres as part of a growth monitoring programme. RESULTS: Nationally, a small but significant increase in levels of underweight takes place during January-March. Participation in growth monitoring also varies seasonally and could account for the increase observed. No evidence of seasonal variation in underweight prevalence is found in the majority of districts studied, although 11 of the districts showed a similar pattern to the national data set. This peak in the incidence of poor nutritional status also coincides with the period of food scarcity before harvest, which is also associated with higher prevalence of diarrhoea and malaria. No differences in seasonality of under-nutrition were found between districts with predominantly subsistence agriculture and those with more commercial forms of agriculture. CONCLUSIONS: Seasonal variation in child weight-for-age exists in some parts of Zimbabwe, but its effects on cross-sectional prevalence studies are likely to be small. There are no readily discernible differences between areas that show evidence of seasonality in levels of underweight and those that do not. PMID- 11415483 TI - Habitual diet in four populations of African origin: a descriptive paper on nutrient intakes in rural and urban Cameroon, Jamaica and Caribbean migrants in Britain. AB - BACKGROUND: The prevalence of chronic diseases is increasing in West Africa, the Caribbean and its migrants to Britain. This trend may be due to the transition in the habitual diet, with increasing (saturated) fat and decreasing fruit and vegetable intakes, both within and between countries. OBJECTIVE: We have tested this hypothesis by comparing habitual diet in four African-origin populations with a similar genetic background at different stages in this transition. DESIGN: The study populations included subjects from rural Cameroon urban Cameroon Jamaica and African-Caribbeans in Manchester, UK all aged 25-74 years. Habitual diet was assessed by a food-frequency questionnaire, specifically developed for each country separately. RESULTS: Total energy intake was greatest in rural Cameroon and lowest in Manchester for all age/sex groups. A tendency towards the same pattern was seen for carbohydrates, protein and total fat intake. Saturated and polyunsaturated fat intake and alcohol intake were highest in rural Cameroon, and lowest in Jamaica, with the intakes in the UK lower than those in urban Cameroon. The percentage of energy from total fat was higher in rural and urban Cameroon than in Jamaica and the UK for all age/sex groups. The opposite was seen for percentage of energy from carbohydrate intake, the intake being highest in Jamaica and lowest in rural Cameroon. The percentage of energy from protein increased gradually from rural Cameroon to the UK. CONCLUSIONS: These results do not support our hypothesis that carbohydrate intake increased, while (saturated) fat intake decreased, from rural Cameroon to the UK. PMID- 11415484 TI - Dietary intake and nutritional status of women and pre-school children in the Republic of the Maldives. AB - OBJECTIVE: Malnutrition and poor nutritional status among children are common problems in the Republic of Maldives, a small island nation in the Indian Ocean. The aim of this study was to determine possible macro- and micronutrient deficiencies in the traditional Maldivian diet. DESIGN: In five atolls, 333 women with children aged between 1 and 4 years who were no longer breast-fed were interviewed, using a 24-hour recall. Additionally, the weights and heights of both the women and children were measured, and blood samples from 15 women were collected for measurements of vitamins A and E, beta-carotene, homocysteine, cholesterol and haemoglobin. RESULTS: Of the women, 22% had a body mass index (BMI) below 18.5. Of the children, 41% were stunted, 14% were wasted and 51% were underweight. The women's and children's diets were sufficient in protein (14%) and carbohydrates (67%) but deficient in fat, which contributed only 19% to the total energy intake. Consumption of dietary substances that depend on vegetable and fruit intake (e.g. beta-carotene, vitamin C, dietary fibre and folic acid) was low. The low intake of beta-carotene was underlined by low plasma concentration. The estimated iron intake was low, although blood haemoglobin levels were normal. CONCLUSIONS: Marginal nutritional status and marginal malnutrition are due to low fat intake and selected micronutrient deficiency. Higher intakes of locally available vegetables and fruits and fat (especially for children) on a regular basis might reverse the deficits documented on the atolls. PMID- 11415485 TI - Dietary intake and nutritional status of US adult marijuana users: results from the Third National Health and Nutrition Examination Survey. AB - OBJECTIVES: This study examined dietary intakes and nutritional status of marijuana users and non-current marijuana users in US adults aged 20 to 59 years. METHODS: We used data from the Third National Health and Nutrition Examination Survey (NHANES III), 1988-1994. Information on self-reported drug use, including marijuana, was obtained as part of the NHANES III physical examination component. Nutritional status was assessed using height and weight, and blood biochemistries. Dietary intake was assessed using a 24-hour recall and a food frequency questionnaire. RESULTS: Among adults 20-59 years of age, 45% reported ever having used marijuana in their lifetime. A total of 8.7% reported using marijuana in the past month. Current marijuana users had higher intakes of energy and nutrients than non-current marijuana users; however, body mass index (BMI) was slightly lower. We found higher cigarette-smoking rates and higher consumption of sodas and alcohol, specifically beer, among marijuana users than among non-current marijuana users. Marijuana users also consumed more sodium, fewer fruits, and more pork, cheese, and salty snacks. Nutritional status, using indicators of serum nutrients, serum albumin, haematocrit and haemoglobin, was similar between marijuana users and non-current marijuana users. Serum carotenoid levels were lower among marijuana users, independent of cigarette smoking. CONCLUSION: Dietary intake was different among marijuana users than among non current marijuana users. Although overall nutritional status was similar, serum carotenoid levels were lower in marijuana users. The long-term health effects of these differences in marijuana users deserve careful consideration in future research. PMID- 11415486 TI - Identification of groups who report similar patterns of diet among a representative national sample of British adults aged 65 years of age or more. AB - OBJECTIVES: Using a national representative sample to identify groups within the UK male and female population over 65 years who report similar patterns of diet. DESIGN: National representative dietary survey, using 4-day weighed dietary records of men and women aged over 65 years old and living in private households in Great Britain in 1994-1995. Cluster analysis was used to aggregate individuals into diet groups. SETTING: United Kingdom. SUBJECTS: 558 women and 539 men. MAIN OUTCOME MEASURES: Consumption of predefined food groups, nutrient intakes, socio economic, demographic and behavioural characteristics. RESULTS: Three large clusters comprising 86% of the male population and three large clusters comprising 83% of the female population were identified. Among men, the most prevalent cluster was a "mixed diet" with elements from a traditional diet and some elements from a healthy diet (48% of the male population); the second was a "healthy diet" (21% of the male population); and the third was a "traditional diet high in alcohol" (17% of the male population). Among women, the most prevalent diet was a "sweet traditional diet" (33% of the female population); the second was a "healthy diet" (32% of the female population); and the last was a "mixed diet" with elements of the traditional diet and the healthy diet (18% of the female population). There were important differences in average nutrient intakes, socio-demographic and behavioural characteristics across these diet clusters. CONCLUSIONS: Cluster analysis identified three diet groups among men and three among women. These differed not only in terms of reported dietary intake, but also with respect to their nutrient content, social and behavioural profile. The groups identified could provide a useful basis for health promotion based upon the diet clusters. PMID- 11415487 TI - Does the condition of the mouth and teeth affect the ability to eat certain foods, nutrient and dietary intake and nutritional status amongst older people? AB - OBJECTIVES: To assess how the dental status of older people affected their stated ability to eat common foods, their nutrient intake and some nutrition-related blood analytes. DESIGN: Cross-sectional survey part of nation-wide British National Diet and Nutrition Survey: people aged 65 years and older. Data from a questionnaire were linked to clinical data and data from four-day weighed dietary records. Two separate representative samples: a free-living and an institutional sample. Seven-hundred-and-fifty-three free-living and 196 institution subjects had a dental exam and interview. RESULTS: About one in five dentate (with natural teeth) free-living people had difficulty eating raw carrots, apples, well-done steak or nuts. Foods such as nuts, apples and raw carrots could not be eaten easily by over half edentate (without natural teeth but with dentures) people in institutions. In free-living, intakes of most nutrients and fruit and vegetables were significantly lower in edentate than dentate. Perceived chewing ability increased with increasing number of teeth. Daily intake of non-starch polysaccharides, protein, calcium, non-haem iron, niacin, vitamin C and intrinsic and milk sugars were significantly lower in edentate. Plasma ascorbate and retinol were significantly lower in the edentate than dentate. Plasma ascorbate was significantly related to the number of teeth and posterior contacting pairs of teeth. CONCLUSIONS: The presence, number and distribution of natural teeth are related to the ability to eat certain foods, affecting nutrient intakes and two biochemical measures of nutritional status. PMID- 11415488 TI - Nutrition and health in an adult urban homeless population in Germany. AB - OBJECTIVE: To assess the association between nutrition and health in an adult urban homeless population. DESIGN: Cross-sectional--nutritional state (body mass index (BMI), triceps skinfold (TSF), upper arm circumference), dietary habits (food frequency), socio-demographic data and self-stated diseases were assessed. SETTING: Four sites for homeless people in Kiel and Hamburg, Germany. SUBJECTS: Sample of 75 homeless people (60 males, 15 females) aged 19-62 years. RESULTS: A lack of food was not found in the majority of the homeless. Seventy-six per cent of the study population showed a normal dietary pattern. Critical food groups were fresh fruit and vegetables, rice and noodles. However, 52 or 29% of the homeless were malnourished (i.e. they were below the 25th or 5th percentile of arm muscle area). In addition, 22.7% of the homeless were obese (i.e. BMI>30 kg m 2 and/or TSF>90th percentile). Almost two-thirds of the population suffered from at least one chronic disease (prevalence of nutrition-related disorders 33.3%, gastrointestinal disorders 32.0%, dental diseases 22.7%, psychiatric disorders 18.7%, wasting diseases 6.7%). Smoking (prevalence rate 82%), drinking alcohol (51%) and drug abuse (20%) were frequent among homeless people. Food intake was not related to nutritional state, the prevalence of chronic diseases or addiction habits. By contrast, a poor nutritional state was associated with drug abuse and the prevalence of wasting diseases. CONCLUSION: Prevention of nutritional problems should be directed to health-related problems such as the prevention or treatment of chronic diseases and addiction habits. PMID- 11415489 TI - The consumption of rye bread and white bread as dimensions of health lifestyles in Finland. AB - OBJECTIVE: The aim of this research was to describe the variation in bread consumption within social classes and to link this consumption to health-related lifestyles in Finland from 1978 until 1998. DESIGN: A cross-sectional survey on health-related behaviour and socio-demographic factors has been conducted annually since 1978. SETTING: Mailed questionnaire. SUBJECTS: A random sample of 5000 Finns aged 15-64 years has been drawn annually. The response rate has varied from 84% to 68%. RESULTS: The consumption of rye bread decreased, but among the female population a slight increase appeared in the 1990s. The consumption of rye bread was associated with a low educational level and a rural place of residence. White bread was consumed less than was rye bread. White bread was consumed more frequently by the less educated in urban areas. Contrary to white bread, the consumption of rye bread was not associated with smoking, exercise or alcohol consumption. CONCLUSIONS: The traditional place of rye bread in the Finnish dietary pattern has remained rather constant. White bread consumption has been associated with an unhealthy lifestyle. As a result, those Finns who are concerned about their health avoid white bread but seem not to associate rye bread with a healthier lifestyle. In Finland, rye bread has a different image to the image of whole-grain or dark brown bread in many other western European countries. PMID- 11415490 TI - Vitamin supplement use and breast cancer in a North Carolina population. AB - OBJECTIVE: Laboratory data suggest that several different vitamins may inhibit the growth of mammary cancers, however epidemiologic data on the relationship between vitamin supplement use and breast cancer are inconsistent. We examined the association between self-reported vitamin supplement use and breast cancer among black women and white women. DESIGN AND SETTING: The data came from a population-based, case-control study conducted in North Carolina between 1993 and 1996. Logistic regression models were used to calculate adjusted odds ratios (ORs) for breast cancer associated with the use of multivitamins or individual vitamin supplements. SUBJECTS: Eligible cases were aged 20 to 74, and approximately 40% of the study population were black women. The analyses included 861 cases and 790 controls. RESULTS: Among all women, there was little evidence for an association between any vitamin supplement and breast cancer. Modest inverse associations were observed among white women for use of multivitamins 95% confidence interval (CI): 0.59-1.12), vitamin C 95% CI: 0.54-1.14) and vitamin E 95% CI: 0.49-1.13). There was no evidence that vitamin supplements reduced the risk of breast cancer among black women. CONCLUSIONS: This study provided very limited support for the hypothesis that vitamin supplements may reduce the risk of breast cancer. Although dietary factors are likely an important influence in breast cancer aetiology, reductions in risk are most likely to be achieved through dietary modification rather than through vitamin supplementation. PMID- 11415491 TI - Serum carotenoids as biomarkers of fruit and vegetable consumption in the New York Women's Health Study. AB - OBJECTIVE: To investigate the usefulness of serum carotenoids as biomarkers of fruit and vegetable consumption. DESIGN: : Reproducibility study on three repeat measurements of serum carotenoids. Correlation analysis of carotenoids and dietary food intake, and regression analysis of potential predictive parameters for serum carotenoid levels. SETTING: New York, USA. SUBJECT: : Women participating in the New York Women's Health Study, a prospective study of sex hormones, diet and breast cancer. Forty-eight women with three repeat blood samples and 302 women having a blood sample and a dietary history questionnaire (including 47 subjects from the reproducibility study). RESULTS: Serum carotenoid concentrations were highly reproducible between one- and two-year repeat samples. Estimated fruit and vegetable consumption was positively correlated with serum carotenoid concentrations but correlation coefficients were low. Consumption of fruit was predictive for serum levels of beta-carotene, alpha-carotene and beta cryptoxanthin, while vegetable consumption was predictive for serum beta carotene, lutein, zeaxanthin and lycopene. Serum concentrations of cholesterol and triglycerides were predictive for serum carotenoids but adjustment for their levels had little or no influence on the correlation between fruit and vegetable consumption and serum carotenoid concentrations. CONCLUSIONS: One single serum measurement of alpha-carotene, beta-carotene and lutein can accurately rank subjects according to their usual serum level. Serum concentrations, however, correlate only moderately with estimated dietary intake of fruits or vegetables and should therefore be used with caution as biomarkers of fruit and vegetable intake. PMID- 11415492 TI - Using isothiocyanate excretion as a biological marker of Brassica vegetable consumption in epidemiological studies: evaluating the sources of variability. AB - OBJECTIVE: Brassica vegetable consumption (e.g. broccoli) leads to excretion of isothiocyanates (ITC) in urine. We evaluated the consistency of ITC as a biomarker for dietary Brassica vegetable consumption across the types of vegetables and methods of preparation used in Western societies, and across consumption levels. DESIGN: A single-armed behavioural intervention with duplicate baseline assessment and post-intervention assessment. Urinary ITC excretion and estrogen metabolites were measured from 24-hour urine samples. Dietary intake was measured by a 24-hour recall. SETTING: The behavioural intervention facilitated daily Brassica intake among participants by providing peer support, food preparation instruction, guided practice in a teaching kitchen, and other information. SUBJECTS: Thirty-four healthy free-living postmenopausal women who recently had a negative screening mammogram at the University of Massachusetts Medical Center. RESULTS: Urinary ITC excretion and total Brassica intake followed the same pattern over the intervention. The ITC biomarker significantly predicted Brassica intake when Brassica consumption averaged about 100 g day-1, but not when Brassica consumption averaged about 200 g day-1. Urinary ITC levels were somewhat higher when more raw vegetables were consumed as compared to lightly cooked vegetables, while the types of Brassica consumed appeared to have only a small, non-significant effect on urinary ITC levels. CONCLUSION: Urinary ITC excretion would be a good exposure biomarker among populations regularly consuming a vegetable serving/day, but may be less accurate among populations with greater intake levels or a wide range of cooking practices. PMID- 11415493 TI - Nutritional methods in the European Prospective Investigation of Cancer in Norfolk. AB - OBJECTIVE: To describe methods and dietary habits of a large population cohort. DESIGN: Prospective assessment of diet using diet diaries and food-frequency questionnaires, and biomarkers of diet in 24-h urine collections and blood samples. SETTING: Free living individuals aged 45 to 75 years living in Norfolk, UK. SUBJECTS: Food and nutrient intake from a food-frequency questionnaire on 23 003 men and women, and from a 7-day diet diary from 2117 men and women. Nitrogen, sodium and potassium excretion was obtained from single 24-h urine samples from 300 individuals in the EPIC cohort. Plasma vitamin C was measured for 20 846 men and women. RESULTS: The food-frequency questionnaire (FFQ) and the food diary were able to determine differences in foods and nutrients between the sexes and were reliable as judged by repeated administrations of each method. Plasma vitamin C was significantly higher in women than men. There were significant differences in mean intake of all nutrients measured by the two different methods in women but less so in men. The questionnaire overestimated dairy products and vegetables in both men and women when compared with intakes derived from the diary, but underestimated cereal and meat intake in men. There were some consistent trends with age in food and nutrient intakes assessed by both methods, particularly in men. Correlation coefficients between dietary intake assessed from the diary and excretion of nitrogen and potassium in a single 24-h urine sample ranged from 0.36 to 0.47. Those comparing urine excretion and intake assessed from the FFQ were 0.09 to 0.26. The correlations between plasma vitamin C and dietary intake from the first FFQ, 24-h recall or diary were 0.28, 0.35 and 0.40. CONCLUSIONS: EPIC Norfolk is one of the largest epidemiological studies of nutrition in the UK and the largest on which plasma vitamin C has been obtained. Methods for obtaining food and nutrient intake are described in detail. The results shown here for food and nutrient intakes can be compared with results from other population studies utilising different methods of assessing dietary intake. The utility of different methods used in different settings within the main EPIC cohort is described. The FFQ is to be used particularly in pooled analyses of risk from diet in relation to cancer incidence within the larger European EPIC study, where measurement error is more likely to be overcome by large dietary heterogeneity on an international basis. Findings in the UK, where dietary variation between individuals is smaller and hence the need to use a more accurate individual method greater, will be derived from the 7-day diary information on a nested case-control basis. 24-h recalls can be used in the event that diary information should not be forthcoming from some eventual cases. Combinations of results utilising all dietary methods and biomarkers may also be possible. PMID- 11415494 TI - Functional foods: a simple scheme for establishing the scientific validity for all claims. PMID- 11415496 TI - Comparison of two-dimensional echocardiography and pulsed Doppler tissue imaging during dobutamine-atropine stress testing to detect coronary artery disease. AB - In order to compare the diagnostic accuracy of two-dimensional (2-D) echocardiography and pulsed Doppler tissue imaging (pDTI) during dobutamine atropine stress testing (DAST) to detect significant coronary lesions, 41 patients underwent DAST (up to 40 microg/k/min of dobutamine with additional atropine during submaximal heart rate responses) and coronary angiography. Pulsed Doppler tissue sampling of territories corresponding to the left anterior descending (LAD), left circumflex (LCx), and right coronary arteries (RCAs) were performed in the apical four-chamber plus aorta and two-chamber apical views. The measurements were repeated at rest, at low dose (10 microg/k/min), and at peak stress. Pulsed DTI measurements included peak early systolic (Vs), peak early diastolic (Ve), and peak late diastolic (Va) velocities. Harmonic 2-D echocardiography was recorded at rest, low dose, peak stress, and recovery, and compared with pDTI assessment. Positive 2-D echocardiography was considered as infarction or ischemic response. The results were evaluated for the prediction of significant coronary stenosis (50% luminal narrowing). Feasibility of pDTI was 100%, 95%, and 98% for the LAD, the LCx, and RCA territories, respectively. At rest, Vs in territories supplied by arteries with coronary artery disease (CAD) (6.3 +/- 2.0 cm/sec) was not different from those without (6.6 +/- 2.1 cm/sec). Vs increased less in territories supplied by arteries with than without CAD (75 +/- 107% vs 102 +/- 69%, P = NS). Ve was lower in territories with CAD at rest (6.0 +/- 2.1 cm/sec vs 8.2 +/- 3.4 cm/sec, P < 0.0001) and low dose (7.2 +/- 2.1 cm/sec vs 8.8 +/- 3.6 cm/sec, P < 0.01), but similar at peak stress (7.6 +/- 3.5 cm/sec vs 8.1 +/- 3.3 cm/sec). Ve increase was similar in territories with (36 +/ 74%) than without CAD (15 +/- 6 4%). Va was similar at rest and low dose in territories with and without CAD (9.2 +/- 2.7 cm/sec vs 9.1 +/- 2.3 cm/sec and 10.9 +/- 3.1 vs 10.3 +/- 3.6 cm/sec, respectively), but lower at peak stress in territories with CAD (13.3 +/- 4.6 cm/sec vs 15.3 +/- 4.5 cm/sec, P = 0.05). The Va increase was lower in territories with CAD (43 +/- 37% vs 77 +/- 72%, P < 0.05). In a territory-based analysis, a failure to achieve Vs > or =10.5 cm/sec at peak stress in the LAD and LCx, and > or =10.0 cm/sec in the RCA territory, was found to be the more accurate limit to detect CAD in the corresponding arteries: sensitivity (95% confidence intervals): 63% (55-71), P = NS vs 2-D echocardiography: 59% (51-67); specificity 76% (68-84), P < 0.01 vs. 2-D echocardiography: 95% (89-100); and accuracy 69% (63-75), P = NS vs 2-D echocardiography: 76% (70-82). Thus, pDTI is feasible during DAST but not more accurate than 2-D echocardiography for the detection of significant CAD in a territory-based study. PMID- 11415497 TI - Normal coronary angiogram and dobutamine-induced left ventricular obstruction during stress echocardiography: a higher hemodynamic responsiveness to dobutamine. AB - This study assessed the clinical or echographic factors predisposing to dynamic left ventricular obstruction (LVO) during dobutamine echocardiography (DE) in patients with angina-like chest pain but without coronary artery disease (CAD). DE is an effective technique for the noninvasive diagnosis of underlying CAD. During DE, an LVO is not unusual in ischemic patients. METHODS: DE (5-40 microg/kg/min) was performed in 52 consecutive patients with angina-like chest pain and normal coronary angiogram. Mean (standard deviation) age was 61 +/- 10 years (27 men, 25 women). Dobutamine-induced LVO was defined as a new intracavitary flow acceleration of at least 3 msec in the left ventricle. RESULTS: Dynamic LVO was observed during DE in 20 (38%) of the 52 patients and was not related to clinical or baseline echocardiographic parameters. The chronotropic response and the systolic blood pressure during DE were higher in the group with LVO (P < 0.03 and P < 0.05, respectively). Appearance of chest pain during the test was also more frequent when LVO occurred (P < 0.02). CONCLUSION: Dynamic LVO is common during DE in a population of patients with angina-like chest pain without epicardial CAD and is associated with a higher hemodynamic responsiveness to dobutamine. PMID- 11415498 TI - Left ventricular outflow tract obstruction after myocardial infarction due to a hyperdynamic basal septum. AB - Acquired left ventricular outflow (LVOT) obstruction may occur following mitral valve repair or replacement. We describe a case where following a large myocardial infarction the hyperdynamic basilar septum was the cause of LVOT. PMID- 11415499 TI - Left atrial appendage stenosis. AB - We describe an unusual case with an anatomic narrowing at the opening of the left atrial appendage demonstrated by two-dimensional color Doppler and continuous wave Doppler transesophageal echocardiography. PMID- 11415500 TI - Investigation of cystic left ventricular masses with echocardiography and magnetic resonance imaging. AB - We report on two patients who had cystic masses located in the left ventricle associated with findings of congestive heart failure. Both echocardiography and magnetic resonance imaging (MRI) were helpful in identifying features of these masses. Echocardiography was used to guide percutaneous transarterial catheter drainage of the mass in one patient. PMID- 11415501 TI - Traumatic tricuspid insufficiency. PMID- 11415502 TI - Transesophageal three-dimensional echocardiographic assessment of left main coronary artery fistula. AB - We describe a patient with a left main coronary artery fistula with an opening at the superior vena cava-right atrial junction in whom three-dimensional transesophageal echocardiography permitted more accurate assessment of the shape and size of the communication site than two-dimensional transesophageal echocardiography. PMID- 11415503 TI - High intensity focused ultrasound: a method of hemostasis. AB - Acoustic hemostasis is a new field of ultrasound research in which high intensity focused ultrasound (HIFU) is used to induce hemostasis in actively bleeding, injured solid organs and blood vessels. In animal studies, moderate to profuse bleeding from parenchymal and vascular injuries has been arrested within approximately 1 minute of HIFU application, even when a large dose of heparin was administered. Moreover, acoustic hemostasis has shown promise in cauterizing large regions of liver, providing a method for bloodless resectioning of abnormal tissue. Two distinct physical mechanisms of HIFU appear to contribute to hemostasis: (1) a thermal mechanism in which absorption of sound leads to temperature elevations, and (2) mechanical mechanisms (acoustic cavitation) in which gas and vapor-filled voids oscillate with large displacement amplitudes. While the thermal mechanism results in a temperature increase in excess of 70 degrees C in about 1 second, the mechanical mechanism appears to result in structural disruption of tissue and possible release of coagulation-inducing tissue factors. Of utmost importance in further development of HIFU as a clinical tool is targeting and monitoring of the HIFU treatment. We have obtained initial success in integrating HIFU with ultrasound imaging so as to develop an image guided therapy system. Image-guided acoustic hemostasis may provide a valuable method of hemostasis in surgical and prehospital settings with applications in trauma and elective surgery. PMID- 11415504 TI - High intensity focused ultrasound for the treatment of tumors. AB - High intensity focused ultrasound (HIFU) is a technique that was first investigated in the 1940s as a potential method of destroying selective regions within the brain to aid neurobehavioral studies. A beam of ultrasound can be brought to a tight focus at a distance from its source, and if sufficient energy is concentrated within the focus, the cells lying within this focal volume are killed, whereas those lying elsewhere are spared. This is, therefore, a noninvasive method of producing selective and "trackless" tissue destruction in deep-seated targets in the body without damage to overlying tissues. This technique is being investigated in a number of medical fields, including urology, ophthalmology, and oncology. The mechanism for cell killing is mainly thermal in origin. Renewal of interest in this technique is due to the availability of sophisticated medical imaging, which now allows the focal volume to be accurately targeted and also allows the tissue destruction to be monitored during treatment. The burgeoning field of HIFU focused ultrasound surgery (FUS) are reviewed in this article. PMID- 11415505 TI - The use of ultrasound for drug delivery. AB - Ultrasound has been in use for the last three decades as a modality for diagnostic imaging in medicine. Recent studies have shown that nonthermal ultrasound energy could be applied for targeting or controlling drug release. This new concept of therapeutic ultrasound combined with drugs has induced interest in various medical fields. Enhanced effects of thrombolytic agents such as urokinase and TPA with acoustic energy have been demonstrated. Ultrasound transducer-tipped catheters are being developed for treatment of cardiovascular diseases. Other devices with ultrasound transducers implanted in transdermal drug patches are also being evaluated for possible delivery of insulin through the skin. Echo contrast microbubbles could also be used to carry and release genes to various tissues and lesions. Chemical activation of drugs by ultrasound energy for treatment of cancers is another new field recently termed "sonodynamic therapy." Various examples of ultrasound application are under investigation that could lead to revolutionary drug delivery systems of the future. PMID- 11415506 TI - Delivery of drugs with ultrasound. AB - In this article we discuss the potential role of microbubbles, traditionally used as ultrasound contrast agents, for site-specific drug delivery. To reach this goal, microbubbles capable of carrying a drug payload are being developed. To ensure that these microbubbles reach sufficient local concentration at disease sites, specific targeting for diseased tissues can be accomplished using several strategies. These strategies rely on either the intrinsic properties of microbubble shells or conjugation of monoclonal antibodies or other ligands to these shells that recognize antigens expressed in regions of disease. Site specific delivery of antiinflammatory, antineoplastic, and thrombolytic drugs with microbubbles can be further enhanced by the ability to locally destroy microbubbles within an acoustic field, thereby releasing drugs and improving drug efficacy without systemic adverse effects. In the case of thrombi, ultrasound mediated microbubble destruction also may facilitate the process of clot lysis. This review also will consider current limitations and technological advances required for the development of this field. PMID- 11415507 TI - Ultrasound gene therapy: on the road from concept to reality. AB - The promise of gene therapy lies in the potential to ameliorate or cure conditions that are resistant to conventional therapeutic approaches. Progress in vascular and all other fields of gene therapy has been hampered by concerns over the safety and practicality of recombinant viral vectors and the inefficiency of current nonviral transfection techniques. This review summarizes the increasing evidence that exposure of eukaryotic cells to relatively modest intensity ultrasound, within the range emitted by diagnostic transducers, either alone or in combination with other nonviral techniques, can enhance transgene expression by up to several orders of magnitude over naked DNA alone. In combination with the flexibility and excellent clinical safety profile of therapeutic and diagnostic ultrasound, these data suggest that ultrasound-assisted gene delivery has great promise as a novel approach to improve the efficiency of many forms of nonviral gene delivery. PMID- 11415508 TI - Therapeutic ultrasound for gene delivery. AB - In vitro and in vivo studies using perfluorocarbon-exposed sonicated dextrose albumin (PESDA) microbubbles to enhance gene delivery are reviewed. In vitro studies show PESDA binds to oligonucleotides and that ultrasound can be used to deposit these nucleotides. In addition, in vitro studies show that drug release from microspheres is dependent on ultrasound transmission frequency as well as pulsed or continuous application. Early in vivo studies confirm that ultrasound in combination with microbubbles can be used to facilitate gene deposition. However, the role of ultrasound targeting gene delivery remains to be determined. PMID- 11415509 TI - Gene delivery using ultrasound contrast agents. AB - With the human genome product and continuing advances in molecular biology many therapeutic genes have been discovered. In the cardiovascular system, gene therapy has the potential to improve myocardial vascularization and ameliorate congestive heart failure. For successful development of clinical gene therapy, however, effective gene delivery vectors are needed. Ultrasound contrast agents can be used to develop new, more effective vectors for gene delivery. Ultrasound contrast agents lower the threshold for cavitation by ultrasound energy. Using physical properties of microbubbles and coating materials, genetic drugs have been incorporated into ultrasound contrast agents. Gene-bearing microbubbles can be injected IV and ultrasound energy applied to the target region. As the microbubbles enter the region of insonation, the microbubbles cavitate, locally releasing DNA. Cavitation also likely causes a local shockwave that improves cellular uptake of DNA. With transthoracic ultrasound, using commercially available diagnostic ultrasound system and an IV injection of gene-bearing microbubbles, high levels of transgene expression are observed in the insonated region of the myocardium. This new technology using microbubbles and ultrasound for gene delivery merits further study and development. PMID- 11415512 TI - Effect of microsomal triglyceride transfer protein gene variants (-493G > T, Q95H and H297Q) on plasma lipid levels in healthy middle-aged UK men. AB - Microsomal triglyceride transfer protein (MTP) plays a central role in the synthesis of lipoproteins by shuttling lipids between phospholipid membranes to apoB. We have examined the effect of three MTP gene variants, -493G > T, Q95H and H297Q, in 2831 healthy UK middle-aged men. The rare allele frequencies were: 0.25 (95% CI 0.24-0.26) for -493T, 0.054 (95% CI 0.05-0.06) for 95H and 0.32 (95% CI 0.31-0.33) for 297Q. The three variants were in strong allelic association in all pairwise combinations (p < 0.001). None of the variant sites were associated with significant differences in cholesterol, triglyceride, apoB or apoAI levels. When stratified by tertiles of triglycerides for the H297Q variant alone there was a significant effect on apoB levels in men in the top tertile (p = 0.01). Considering the -493G > T and H297Q genotype in combination on baseline levels, individuals with three or four rare alleles had 6.6% higher mean apoB levels compared to the rest (p = 0.007). Therefore, homozygosity for 297Q at higher triglyceride (Tg) levels, or in combination with -493G > T, is associated with a raising effect on apoB levels, suggesting the importance of modest differences in MTP activity in determining hepatic secretion of lipoproteins in healthy men. PMID- 11415513 TI - A clinical, cytogenetic and molecular study of 47 females with r(X) chromosomes. AB - We studied 47 patients with a 45,X/46,X,r(X) karyotype to identify phenotypic differences between these patients and 45,X patients, and to determine whether these differences could be explained by the status of genes within the ring. Only 2 patients had the 'severe' r(X) phenotype, and both were consistent with this resulting from functional disomy of genes normally subject to X inactivation. A further 7 patients also carried active rings but these patients did not have a more severe phenotype than those whose rings were inactivated, probably because their rings were smaller and did not contain the (as yet unidentified) genes whose functional disomy is particularly damaging. Patients with a r(X) did not show clear physical differences when compared with a 45,X series, except for a possible reduction in the frequency of oedema in those whose r(X) had an Xq breakpoint distal to DXS128E, at Xq13.2. Thus some protection from oedema may be provided by the presence of two copies of Xq13.2. PMID- 11415514 TI - Ring-X chromosomes: their cognitive and behavioural phenotype. AB - We tested the cognitive abilities and educational attainments of 47 patients with a ring X chromosome, to evaluate the extent to which these variables correlated with failure of r(X) inactivation and with mosaicism. We found possession of a r(X) chromosome was associated with an increased risk of significant learning difficulties, and with associated behavioural maladjustment, compared with 45,X Turner females. Nearly a third had been educated outside mainstream schools. The proportion of cells in peripheral blood containing an inactivated r(X) chromosome was negatively correlated with nonverbal IQ. The parental origin of the normal chromosome did not appear to affect adjustment or abilities. In a minority of r(X) cases associated with mental retardation, there had been a failure to inactivate the ring, due to loss of the XIST locus. However, failure of X inactivation was not necessarily associated with a severe phenotype. The degree of impairment in IQ depended on the size of the active ring, and hence was proportionate to the number of (as yet unidentified) genes whose functional disomy affected brain development and functioning. PMID- 11415515 TI - Multiple interacting gene products may influence susceptibility to malignant hyperthermia. AB - Malignant hyperthermia (MH) is a potentially lethal disorder triggered in susceptible individuals on exposure to common anaesthetic agents. Crises reflect the consequences of disturbed skeletal muscle calcium homeostasis. MH is an autosomal dominant, genetically heterogeneous trait. Defects in a single major gene have been assumed to determine susceptibility status in individual families. However, in some pedigrees phenotypic and genotypic data are discordant. One explanation, in contrast to the current genetic model, is that susceptibility is dependent upon the effects of more than one gene. Using the transmission disequilibrium test we assessed the involvement of 8 MH candidate loci (RYR1, CACNA1S, CACNA2D1, MHS4 at 3q13.1, MHS6 at 5p, LIPE, DM1, dystrophin) by analysis of data from 130 MH nuclear families. Results suggested that variations in more than one gene may influence MH susceptibility in single families. PMID- 11415516 TI - Northwest African distribution of the CD4/Alu microsatellite haplotypes. AB - We have analysed a linked microsatellite/Alu polymorphism at the CD4 locus (CD4/Alu) in 666 chromosomes from samples of the Iberian Peninsula, Northwest Africa, and West sub-Saharan Africa. The Iberian Peninsula differs from other European populations by its higher levels of haplotype diversity (0.75), and weaker association between the microsatellite allele 90 and Alu(-) chromosomes. These results are explainable by a substantial gene flow from Northwest Africa. Significant geographic clines for the five major haplotypes suggest a south to north migration from sub-Saharan Africa into Northwest Africa. In spite of this, the consistent presence of haplotype 110(-) in this area is congruent with an ancient and autochthonous human presence in Northwest Africa. PMID- 11415517 TI - Genetic diversity at the FMR1 locus in the Indonesian population. AB - We report an analysis of allelic diversity at short tandem repeat polymorphisms within the fragile XA locus in 1069 male volunteers from twelve Indonesian sub populations. An odd numbered allele of DXS548 was found at high frequency in all Indonesian populations. Greater allelic diversity was identified at the loci under study than has been previously reported for an Asian population. These differences distinguish the Indonesian population from all previously reported Asian, European and African populations. A high frequency of small premutation alleles, 4/120 (3.3%, 95% CI 0.9-8.3%), was identified in the Moluccan population of Hiri Island. PMID- 11415518 TI - Linkage analysis of candidate endothelin pathway genes in nonsyndromic familial orofacial cleft. AB - There is good evidence from linkage analysis and mouse model knockouts that the endothelin-1 gene (EDN1) is a good candidate for non-syndromic orofacial cleft (OFC) disease. EDN1 maps to the chromosomal region of the OFC1 locus in 6p23. Therefore we have examined three other candidate genes in the endothelin pathway (ECE1, EDNRA and EDNRB, which map to chromosomes 1, 4 and 13 respectively) in a linkage study of 9 families with OFC, where the disorder is not linked to chromosome 6p23. The total lod score for these 9 multiplex families never exceeded -2.00 and thus our data suggest that EDN1 and related genes are not involved in non-syndromic familial OFC. PMID- 11415519 TI - How selection shapes variation of the human major histocompatibility complex: a review. AB - The nature of polymorphism and molecular sequence variation in the genes of the human major histocompatibility complex (MHC) provides strong support for the idea that these genes are under selection. With the understanding that selection shapes MHC variation new questions have become the focus of study. What is the mode of selection that accounts for MHC polymorphism? Is variation maintained by pathogen pressure or by reproductive mechanisms? Discerning between these requires drawing on information from studies on association between HLA genes and infectious diseases, reproductive success and mating preferences relative to HLA genotypes, and theoretical studies that compare the outcomes of different selection regimes. The pattern that has emerged suggests that several types of selection are plausible for the maintenance of HLA polymorphism. PMID- 11415520 TI - Haplotype analysis of the human alpha2-HS glycoprotein (fetuin) gene. AB - Alpha2-HS glycoprotein (AHSG), which is equivalent to fetuin in other species, is a protein found in human plasma. AHSG is polymorphic with two common alleles and many variants. To examine the intragenic haplotypes and their diversity at this locus, a contiguous genomic DNA sequence (10.3 kb) was analyzed in 20 samples (40 chromosomes), and haplotypes were determined for 309 subjects. Judging from the aligned nucleotide sequences and the conserved amino acid residues comparing human and chimpanzee AHSG, it was concluded that the type 1 allele is probably older and has evolved into four major suballeles. The type 2 allele was generated from one branch of the type 1 allele. AHSG*3 and *5 variants were each found to have a single nucleotide change in exon 7, resulting in the change of an amino acid residue from Arg299 to Cys and from Asp258 to Asn, respectively. It was noted that the AHSG*3 mutation gives rise to an additional cysteine residue, which possibly affects the conformation of the protein. The AHSG gene was found to have a low mutation rate and no apparent recombination events. Furthermore, the detected substitutions were nonhomogeneously distributed at this locus. In particular, four nonsynonymous substitutions were concentrated in the carboxyl terminal domain. PMID- 11415521 TI - Existence of a genetic risk factor on chromosome 5q in Italian coeliac disease families. AB - Coeliac disease (CD) is a malabsorptive disorder of the small intestine resulting from ingestion of gluten. The HLA risk factors involved in CD are well known but do not explain the whole genetic susceptibility. Several regions of potential linkage on chromosomes 3q, 5q, 10q, 11q, 15q and 19q have already been reported in the literature. These six regions were analyzed with the Maximum Lod Score method on a dense set of markers. A new sample of 89 Italian sibpairs was available for study. There was no evidence for linkage for any of the regions tested, except for chromosome 5q. For this region, our data, as well as a sample of 93 sibpairs from our first genome screen (Greco et al. 1998), are compatible with the presence of a risk factor for CD with a moderate effect. PMID- 11415522 TI - The phylogeography of Y chromosome binary haplotypes and the origins of modern human populations. AB - Although molecular genetic evidence continues to accumulate that is consistent with a recent common African ancestry of modern humans, its ability to illuminate regional histories remains incomplete. A set of unique event polymorphisms associated with the non-recombining portion of the Y-chromosome (NRY) addresses this issue by providing evidence concerning successful migrations originating from Africa, which can be interpreted as subsequent colonizations, differentiations and migrations overlaid upon previous population ranges. A total of 205 markers identified by denaturing high performance liquid chromatography (DHPLC), together with 13 taken from the literature, were used to construct a parsimonious genealogy. Ancestral allelic states were deduced from orthologous great ape sequences. A total of 131 unique haplotypes were defined which trace the microevolutionary trajectory of global modern human genetic diversification. The genealogy provides a detailed phylogeographic portrait of contemporary global population structure that is emblematic of human origins, divergence and population history that is consistent with climatic, paleoanthropological and other genetic knowledge. PMID- 11415523 TI - Mitochondrial DNA variability in Russians and Ukrainians: implication to the origin of the Eastern Slavs. AB - In order to investigate the origin of the Eastern Slavs, mitochondrial DNA (mtDNA) sequence variation was examined in Russians and Ukrainians by hypervariable segment I (HVS I) sequencing and restriction analysis of the haplogroup-specific sites. No significant differences were found for Russians and Ukrainians when compared to other Europeans - in fact, they fall within the range of gene diversity seen throughout Europe and exhibit the unimodal pattern of pairwise sequence differences. Moreover, HVS I sequences in the Russians and Ukrainians are similar or identical to those found in eastern and western European populations. Despite the small genetic distances between Europeans, phylogenetic analysis reveals a considerable heterogeneity of Eastern Slavonic populations - they do not cluster together onto a phylogenetic tree. Analysis of distribution of rare HVS I types shared between populations of Eastern Slavs and other West Eurasians has shown that Russians share rare haplotypes mainly with Germans and Finno-Ugric populations. Of these, subhaplogroup H1 sequence types, which are defined by different combinations of nucleotides 16192T, 16294T, 16304C, 16311C and 16320T, are found predominantly in common between Russians and German-speaking populations. The data obtained allow us to conclude that the Slavonic migrations in early Middle Ages from their putative homeland in central Europe to the east of Europe were accompanied mostly by the same mtDNA types characteristic for the pre-Slavonic populations of eastern Europe. PMID- 11415524 TI - Genetic crossover interference in the human genome. AB - Positive crossover interference refers to the phenomenon that the occurrence of a crossover reduces the probability of another crossover in its vicinity. There have been studies reporting the presence of positive interference in humans. Some studies have also found evidence suggesting within and between chromosomal interference heterogeneity on some of the chromosomes. However, there has been no systematic study of interference and interference heterogeneity in the whole human genome, using pedigree data without first inferring crossovers. In this paper, we studied the Chi-square interference model and other models extensively to compare the relative performance of each of these models for accounting for interference and measuring strength of interference. Our results showed that the Chi-square model consistently fitted the data well and provided easily interpretable estimates of interference strength. The Chi-square model was then used to study interference and interference heterogeneity within and between chromosomes. We found strong evidence of positive interference in the whole human genome. Our results also indicated that the level of interference was fairly constant in most parts of the genome, but there was some evidence suggesting that the levels of interference for two of the chromosomes were different from the rest. We also found evidence of within chromosomal interference heterogeneity for several of the chromosomes. PMID- 11415525 TI - Use of an artificial neural network to detect association between a disease and multiple marker genotypes. AB - Single nucleotide polymorphisms (SNPs) are very common throughout the genome and hence are potentially valuable for mapping disease susceptibility loci by detecting association between SNP markers and disease. However as SNPs are biallelic they may have relatively little power in association studies compared with the information that would be obtainable if marker haplotypes were available and could be used efficiently. Modelling the evolutionary events leading to linkage disequilibrium is very complex and many methods that seek to use information from multiple markers simultaneously need to make simplifying assumptions and may only be applicable when marker haplotypes, rather than genotypes, are available for analysis. We explore the properties of a simple application of a standard artificial neural network to this problem. The pattern recognition properties of the network are used in the hope that marker haplotypes implicit in the genotypes will differ between cases and controls in a way which will lead to the network being able to classify the subjects correctly, according to their marker genotype. This method makes no assumptions at all regarding population history or the marker map, and can be applied to genotypes, as would be available from a simple case-control sample, without any need to determine haplotypes. Through application to data simulated under a very wide range of assumptions we show that such an analysis produces a useful augmentation in power above that which would be achieved by testing each marker individually, in particular when more than one mutation has occurred in a disease gene at different points in evolution. The application of neural networks to such problems shows considerable promise and further work could usefully be directed towards optimising the design and implementation of such networks. PMID- 11415526 TI - Recent advances in lumbar spinal mechanics and their clinical significance. AB - Of the many problems associated with low back pain, those which are most amenable to biomechanical investigation are identified. Recent advances in lumbar spinal mechanics are then reviewed in five sections dealing with mechanical function, mechanisms of failure, movements in vivo, loading in vivo, and the biological consequences of mechanical loading. The discussion suggests that mechanical fatigue damage may frequently be the underlying cause of low back pain, even when degenerative changes are evident in the tissues, and the review ends by suggesting some priority areas for future research. PMID- 11415527 TI - Relationships between one-handed force exertions in all directions and their associated postures. AB - Photographs were taken of subjects exerting in specified directions with one hand on the handle of a triaxial force measurement system. The applied forces were recorded and posture analysis was undertaken to investigate relationships between three-dimensional force exertion and posture. The postural stability diagram, which in previous studies has been applied to fore-and-aft exertions, was applied to the vertical plane containing the manual force vector and to the horizontal plane. The vertical plane analysis provided an insight into postures associated with weak and strong exertion. The horizontal plane analysis emphasized the importance of developing torque as well as thrust at the foot base in order to exert laterally directed forces. Exertions involving a right or left component were associated with a horizontal moment at the feet of the order of 50 Nm. This moment is an important factor in the demands made upon the body during asymmetrical exertion, and the mechanisms for achieving it deserve further investigation. RELEVANCE: Exertion is not normally restricted to the sagittal plane. The approach adopted in this paper gives an insight into how body deployment relates to the direction and magnitude of exertion. Biomechanical models of asymmetric exertion should reflect the principles that have emerged. PMID- 11415528 TI - Biomechanical properties of normal tendons, normal palmar aponeuroses, and tissues from patients with Dupuytren's disease subjected to elastase and chondroitinase treatment. AB - Normal tendons, normal palmar aponeuroses and palmar aponeuroses from patients with Dupuytren's disease were subjected to elastase or chondroitinase treatment. Young's modulus was derived from the linear portion of stress-strain graph. It showed the lowest value for the apparently normal palmar aponeuroses and the highest value for tendon samples. Elastase treatment caused an increase of extensibility and a reduction of Young's modulus of normal palmar aponeuroses and tendons, but not of contracture bands. In normal tendons, normal palmar aponeuroses and apparently normal palmar aponeuroses residual strain and hysteresis loop increased significantly as a linear function of the amount of digested elastin. In contrast these biomechanical parameters were not affected significantly in contracture bands. In normal and apparently normal areas incubation with chondroitinase ABC resulted in a significant increase of residual strain and, as opposed to elastase, a decrease of normalized hysteresis loop. In contracture bands, however, these biomechanical parameters remained unchanged. RELEVANCE: The increasing evidence of a correlation between morphological changes of palmar elastin and ground substance with the progress of Dupuytren's disease emphasizes the need to determine the relative importance of these connective tissue components for the pathogenesis of Dupuytren's disease. PMID- 11415529 TI - Comparison of 2-dimensional and 3-dimensional rearfoot motion during walking. AB - The purpose of this study was to investigate the validity of two-dimensional (2 D) analysis with respect to three-dimensional (3-D) analysis for recording rearfoot motion during walking. The results of this study indicated that rearfoot motion measured with either 2-D or 3-D analysis is essentially the same for the initial 60% of the stance phase. There was no significant difference between the two methods on variables typically measured in studies dealing with rearfoot motion. RELEVANCE: The assessment of rearfoot motion during walking in a clinical situation is frequently desirable yet the high cost of equipment and time required for 3-D analysis makes it unreasonable or impractical. The results of this study show that 2-D analysis can be used to measure rearfoot motion accurately and therefore is feasible for use in the clinic. PMID- 11415530 TI - Correlation between physical activity and the gait characteristics and ankle joint flexibility of the elderly. AB - The correlation between the type, duration and intensity of physical activity on the gait characteristics and the range of active ankle joint motion of the elderly were investigated in this study. Three-dimensional (3D) kinetic and kinematic assessments were performed on the normal walking patterns of 59 healthy elderly male and female subjects (aged 60-79 years). Gait analysis was performed using a high-speed video-based (3D) motion analysis system with synchronized ground-reaction force measurements. The maximal active range of motion (RoM) of the ankle joint complex of each subject was determined using an external six degree-of-freedom flexibility assessment device. Physical activity levels were classified based on energy expenditure requirements. In general the results suggest that habitual level of physical activity did not have a significant effect on the kinetic or kinematic variables during walking, or the maximal ankle joint range of motion. Differences in ankle joint RoM and gait variables were found based on gender. No distinct benefit with respect to ankle joint range of motion or gait characteristics was provided with participation in higher-energy intensive physical activities in comparison to physical activities requiring low to moderate energy expenditures. It is speculated that the benefits of physical activity may be more pronounced in activities which demand rapid muscular strength and control movements such as recovering from a fall or obstacle avoidance. RELEVANCE: Participation in low-energy intensity activities provides the same benefits to ankle joint flexibility and locomotion, without the added risk of injury associated with high-energy intensity sports. This finding has important implications with respect to physical activity programmes for the elderly with an objective to maintain or improve independent mobility. PMID- 11415531 TI - On the mechanical function of tendon. AB - A mesoscopic approach is followed for mathematical modelling of the specific deformation properties of tendon. The approach starts from our general concept of modelling mechanical behaviour of heterogeneous media and assumes that the structure of tendon is optimized in such a way that it enables its adjacent muscle to work with a constant performance in the course of increasing loading (acting like a gearbox in a car). The model based on this assumption gives results that are in a very good accordance with observed properties of tendons. Clinical experience reveals that if this function of tendon is violated pathological changes appear in the respective muscle. RELEVANCE: Clarification and mathematical modelling of the mechanical function of tendon is of intellectual interest in its own right, but it is important also for cautioning surgeons against unnecessary violation of this function, and for tissue engineering aspects if tendon must be replaced. PMID- 11415532 TI - Omnidirectional assessment of one-handed manual strength at three handle heights. AB - Maximal right-handed omnidirectional strengths of eleven males were measured at three handle heights of 1.5 m, 1.0 m and 0.5 m using the Tri-Axial Force measurement System. The postures of the subjects were constrained by preventing rotation of the footbase in order to ensure that the forward-backward and leftward-rightward axes of the subject remained constant relative to the measurement system. A two-way repeated measures analysis of variance revealed highly significant differences in maximal strengths between the three heights and the 614 directions of exertion measured. A highly significant interaction was found between height and direction. At all three hand heights a restricted area of great strength was found in the direction of upward and almost directly forward from the subject. A second, less powerful, but more widespread area of high strength was also found at each height. At the 1.5-m height the peak was in the backward and almost vertically downward area. At the 1.0-m height the peak was in the area of slightly downward and largely backward. At the 0.5-m height the peak was to the right of upward and backward. RELEVANCE: Very few real-life manual exertions involve forces which are purely in the sagittal, forward backward plane, with lateral components of force usually being involved as well. The data presented extend the knowledge base about male one-handed strengths. PMID- 11415533 TI - Measurement of creep strain of flexor tendons during low-force high-frequency activities such as computer keyboard use. AB - The aim of this study was to measure tendon strain during low-force, high frequency activities such as computer keyboard use. Prior to creep strain testing an estimate of flexor tendon force during keyboard use was made. Tendon force was measured indirectly by comparing electromyographic activity of the flexor and extensor digitorum muscles in five human volunteers for various hand activities. Results of the electromyographic study showed that flexor tendon forces during keyboard use may be as high as 60 N. Sixty eight flexor digitorum tendons from 17 fresh-frozen cadaver hands were used for the creep strain tests. Three loading conditions (static, 1 Hz cyclic, (1/4) Hz cyclic) and four load levels (10, 20, 50, 100 N) were used. Results of the creep study showed that for a flexor tendon force of 60 N the total strain of the tendon would be approximately 1.8%. This does not appear to be enough strain to cause permanent damage to the tendon according to current cumulative strain models. RELEVANCE: Recent studies have shown an increase in hand and wrist tendinitis among computer users. At the present time the aetiology is unknown, but risk factors commonly associated with tendinitis are hand forces, wrist postures, and frequency of finger movements. This research looks at tendon creep due to repeated force as a possible aetiology. PMID- 11415534 TI - Oblique abdominal muscle activity in standing and in sitting on hard and soft seats. AB - The activity of the oblique abdominal muscles was investigated with the trunk in unconstrained, symmetrical and static postures. Electromyographic recordings in six healthy subjects revealed that in all subjects the activity of both the internal and the external obliques is significantly higher in unconstrained standing than in supine posture. Activity of the internal oblique was higher than that of the external oblique abdominal. The sacrospinal, gluteus maximus and biceps femoris muscles showed practically no activity in unconstrained erect posture. During unconstrained sitting both oblique abdominals are active. In most subjects the activity of the oblique abdominals was significantly smaller when sitting on a soft car seat than when sitting on an office chair with a hard seat. The possibility is discussed that contraction of the oblique abdominals in unconstrained standing and sitting may help in stabilizing the basis of the spine and particularly the sacroiliac joints. During standing and sitting the oblique abdominal muscles apparently have a significant role in sustaining gravity loads. RELEVANCE: Back pain and pelvic pain are often experienced in prolonged standing and sitting postures. In these postures the oblique abdominals are shown to be active. The present study gains clinical significance by the studies showing relatively small oblique abdominal muscle strength in patients with low back pain. A soft seat may be helpful in treatment and prevention, because it substitutes oblique abdominal muscle activity. PMID- 11415535 TI - Kinematic and kinetic comparison of downhill and level walking. AB - Kinematic and kinetic data were collected from 12 healthy subjects whilst they performed both downhill and level walking at a controlled cadence. A ramp of 6 m length and a gradient of -19% was used for downhill walking and this incorporated the same force platform that was used for level walking. Planar net joint moments and mechanical power at the ankle, knee, and hip joints were calculated for the sagittal view using force platform and video records based on standard inverse dynamics procedures. On the basis of differences in ankle, knee, and hip joint kinematics the ankle joint was seen to compensate for the gradient at push off and during the swing, the knee joint from early stance through until early swing phase, and the hip joint from early swing through until the early stance phase. The major differences in joint moments and muscle mechanical power were seen in the knee and ankle joint. Whereas peak moments and muscle power were much higher for downhill walking in the knee joint, these measures were significantly smaller at the ankle joint. Hip joint moments and muscle power estimates were only slightly larger for downhill walking. These data explain well the problems that patients with patellofemoral pathology and anterior cruciate ligament (ACL) deficiency encounter with downhill walking, and the muscle soreness experienced by mountain trekkers. RELEVANCE: Biomechanical estimates of musculoskeletal loadings in gait are invariably derived from laboratory studies of walking over a level surface. In this study comparisons were made between downhill and level walking in order to appreciate more fully the increased loadings on the lower extremity under more stressful but not atypical conditions. The data so derived provide the necessary basis for the prediction of loadings on specific muscle/joint structures and can serve as a foundation for exercise prescription with patients recovering from injury or orthopaedic surgery. PMID- 11415536 TI - A videofluoroscopy method for optical distortion correction and measurement of knee-joint kinematics. AB - Image distortion in video and image intensifier X-ray systems requires appropriate distortion correction methods to obtain accurate biomechanical quantitative measurements for joint kinematics applications. This paper presents an algorithm for coordinate reconstruction and distortion correction using a modified polynomial method. This algorithm was used for the measurement of patellar tendon moment arm, tibial plateau-tibial axis angle and patellar tendon tibial axis angle during knee extension using videofluoroscopy in vivo. These parameters allow the determination of a two-dimensional biomechanical model of the knee for the measurement of muscle and joint forces during dynamic activities. Five males without knee joint injury history participated in the study. The mean measurement error obtained using an image intensifier-video system was 0.246 +/- 0.111 mm over a 180-mm x 180-mm field of view. The mean maximum patellar tendon moment arm was 39.87 mm at 44.9 degrees of knee flexion. The patellar tendon-tibial plateau angle was 112.9 degrees at full extension and decreased linearly to 87.6 degrees at 90 degrees of knee flexion. The mean angle between the tibial plateau and the tibial long axis was 84.8 degrees. Applications of the method include motion analysis using video and X-ray fluoroscopy systems with non-linear distortion problems. RELEVANCE: Accurate measurement of anatomical parameters from videofluoroscopy systems is important for the determination of joint biomechanical models and measurement of muscular and joint forces. PMID- 11415537 TI - Quantitative analysis of anterior cruciate ligament instability. AB - The purpose of this study was to quantify end-feel by evaluating instantaneous stiffness and change in stiffness of the tibiofemoral joint. Normal (n = 26), anterior cruciate ligament (ACL) deficient (n = 6), and posterior cruciate ligament (PCL) deficient (n = 2) subjects were tested using the KT 2000 knee ligament arthrometer. Analogue force and displacement data were digitized, filtered, and calibrated. Force-displacement curves in the region of anterior tibial displacement were approximated using 5th degree polynomial functions. Stiffness and change in stiffness were determined analytically by the first and second derivatives of these functions. Subjects with confirmed ACL deficiency had lower values for stiffness and change in stiffness in the region of the curves believed to be indicative of ACL function, compared to contralateral knees and control subjects. RELEVANCE: First and second derivatives of tibiofemoral force displacement curves give the stiffness and the change in stiffness respectively for anterior displacement of the tibia relative to the femur. This information concerning knee stability in vivo may be useful for diagnosis and follow-up. PMID- 11415538 TI - Exercise, posture, and back pain during pregnancy. AB - One of the expected benefits of exercise programmes for pregnant women is to reduce or prevent back pain by improving posture. The aim of this project was to evaluate the effect of fitness classes for pregnant women on posture and back pain. In this article, postural aspects are reported. Sixty five pregnant volunteers were included in the study, of whom 27 were enrolled in exercise classes designed according to Canadian guidelines and 38 acted as sedentary controls. Posture was assessed every 4 weeks during pregnancy and 4 months postpartum by measuring curvatures of the lumbar and thoracic spines in a standard relaxed standing position from lateral photographs. Laxity of knee ligaments was also monitored using a clinical arthrometer. Weight gain could explain part of lordosis increase during pregnancy but the effect was not very strong. No effect of exercise on posture was detected. RELEVANCE: This study showed that fitness classes for pregnant women designed according to Canadian guidelines had no detectable effect on posture during pregnancy. PMID- 11415539 TI - Exercise, posture, and back pain during pregnancy. AB - In this article the effect of exercise on pregnancy-related back pain is reported. Sixty five pregnant volunteers were included in the study, of whom 27 were enrolled in exercise classes designed according to Canadian guidelines, and 38 acted as sedentary controls. Back pain and functional limitations were assessed every 4 weeks during pregnancy and 4 months postpartum by questionnaires. No significant differences in back pain or functional limitations reported by the two groups were found. The biomechanical factors measured in this study, weight, back posture, and laxity, were unable to explain most of the variation in pain reported. RELEVANCE: This study showed that fitness classes for pregnant women designed according to Canadian guidelines had no detectable effect on back pain during pregnancy. If prevention of pregnancy-related back pain is sought through exercise, design of fitness programmes for pregnant women should be reconsidered. PMID- 11415540 TI - Isokinetic pull strengths in the vertical plane: effects of speed and arm angle. AB - The significance of knowing dynamic (isokinetic) strengths of individuals and the impact of selected individual and work-related factors on these strengths have been addressed in previous works. This follow-up study investigated the effects of pulling speed and angle of the preferred arm on isokinetic strengths of males and females in a vertical plane. The results indicated that speed of pulling has a major influence on strength exertion capabilities of people. Isokinetic strength exertions are maximum in the vertical plane but are smaller than isometric strength exertions in the vertical plane. As expected, men exerted greater force than females. Force exertions in the sitting and standing postures were not appreciably different from each other. RELEVANCE: Knowledge of strength exertion capabilities of individuals is necessary for designing work controls, and in assessing rehabilitative effects of physical therapy programmes. How the speed of motions and orientations of the arm influence strength can assist in designing work which is within the capability of the working population. PMID- 11415541 TI - Surgery simulation analysis of anterior advancement of the tibial tuberosity. AB - The objective of this study is to evaluate the surgical outcome of anterior displacement of the tibial tuberosity (Maquet procedure) for reducing patellofemoral joint contact force. Thein-vivo experimental knee joint geometric data with a biomechanical model was used to do the simulation of the Maquet procedure. Six healthy young adults performed weight-bearing knee flexion extension by ascending a one-step stair. Dynamic X-ray images of the knee were continuously recorded by a video-fluoroscopic system. These X-ray images were analysed on a computerized digitizing system to get the knee joint geometric data. Based on the continuous in-vivo geometric data, computer surgery simulation was studied on six right knees with advancement of 3, 5, 10, 15, and 20 degrees of the patellar tendon insertion. Evaluation of the simulation consequences from a biomechanical view point showed that the Maquet procedure reduced the patellofemoral joint reaction force only up to 20% at 90 degrees of knee flexion angle. The patellofemoral joint reaction force had 50% reduction only when the knee flexion angle less than 20 degrees, and only when the patellar tendon was moved out by 15 or 20 degrees. This represented nearly 1 in. of the anterior displacement of the tibial tuberosity. The results also showed that the Maquet procedure would decrease up to 20% of the force transmission efficiency of the patellofemoral mechanism, which would cause the mechanical consequences of the operation to be only minor at larger knee flexion angles more than 20 degrees. These findings suggest that the Maquet procedure is only favourable to less active or older patients having small knee flexion angle activities. RELEVANCE: Surgical procedure of anterior displacement of the tibial tuberosity has been used to reduce the patellofemoral joint contact force for treatment of symptomatic osteoarthrosis of the patellofemoral joint. In this study the patellofemoral joint reaction force had 50% reduction only when the knee flexion angle was less than 20 degrees, and only when patellar tendon was moved out nearly 1 in. Based on this result, the Maquet procedure is suggested only favourable to less active or older patients having small knee flexion angle activities. PMID- 11415542 TI - Influence of weight and frequency on thigh and lower-trunk motion during repetitive lifting employing stoop and squat techniques. AB - Changes in kinematics as a function of lifting weight and frequency was investigated in sagittal symmetric repetitive lifting. For every lift cycle (lowering and lifting) the motion range between the upright position (0 degrees ) and the maximum angular displacement of the thigh and lower-trunk body segments was recorded. Ten subjects performed five repetitive lifting bouts with different weight/frequency combinations, using both stoop and squat lifting techniques. In total, 6384 lifts were analysed. The lifting weight or frequency did not influence the motion ranges in stoop lifting. In squat lifting the weight lifted did not appear to have any influence on the motion ranges, while the thigh motion range was significantly smaller at lifting frequency of 20 lifts min(-1) than at a frequency of 10. A significant gradual decrease in the thigh motion range and corresponding increase in the lower-trunk motion range were seen for a majority of the subjects during squat lifting at frequency 20. These changes suggest that quadriceps muscle strength is the limiting factor in repetitive squat lifting. Also the variation in motion ranges was greater in squat lifting than in stoop lifting. RELEVANCE: Forestry work involves frequent lifting. However, compliance in using squat lifting technique, which is recommended for safe lifting, is sometimes poor. Fatigue may be one of the determinants for changes in kinematics and choice of technique in lifting tasks. PMID- 11415543 TI - Moments at the L(5)/S(1) joint during asymmetrical lifting: effects of different load trajectories and initial load positions. AB - The purpose of this study was to examine the reaction moments at L(5)/S(1) about the three orthogonal orthopaedic axes on the trunk when performing asymmetrical lifting tasks under six experimental conditions. The first three conditions were three lifting modes referring to different load trajectories (diagonal, extension then twisting, twisting then extension). The next three conditions were three lifts referring to different initial load positions (90 degrees, 45 degrees, 0 degrees ). Ten male volunteers (mean age 23.4 years) participated in the study. The mass of the load was 11.6 kg and two shelves were used: a low shelf of 22 cm from which the load was removed and a high shelf of 80 cm onto which the load was laid down following the manoeuvre. A dynamic 3D multisegment model was used to compute spinal loadings. The maximum reaction moments generated at the joint for the three load trajectories and for the three initial load positions were approximately of the same magnitude. On the other hand, significant differences were observed in the trunk orientation when the maximal moment was generated. Symmetrical trunk positions appeared to be more advantageous because of the reduction in the capability of maximal extension force in asymmetric trunk posture. A possible alternative for avoiding excessive stress on the back in asymmetric trunk posture may be to keep the shoulders parallel to the ground. RELEVANCE: Three-dimensional studies of the resulting moments at L(5)/S(1) when performing lifting activities are not common, particularly when the tasks include muscular exertions in trunk flexion combined with either twisting or lateral bending. This type of analysis is particularly important since asymmetries have been identified as risk factors for low back disorders. The knowledge of reaction moments in relation to postures are essential to understand the mechanism of low back pain. PMID- 11415544 TI - Kinematics of shoulder abduction in the scapular plane. On the influence of abduction velocity and external load. AB - In this paper the kinematics of arm abduction in the scapular plane of 38 healthy experimental subjects is reported. The ratio of the glenohumoral and the scapulothoracic components of the motion were determined and the influence of the abduction speed and of the external load on it were investigated. The investigation of the effect of abduction speed involved all 38 subjects. Each of them performed one slow and one fast abduction. Statistical analysis showed that there were large differences between individuals, but that for any one individual the abduction process is essentially reproducible. For the one individual there is a strong, linear relationship between glenohumeral and scapulothoracic rotation. The slope of the regression of the glenohumeral component on total arm abduction varied from 0.75 to 0.5 with a sample mean of 0.66. It means that only two-thirds of arm abduction occurs in the glenohumeral joint, the remaining third taking place via scapular rotation. In slowly performed abductions the slope of the regression was significantly greater than in the high-speed movements, but differences were very small. In the investigation of the effect of external load, statistical analysis indicated that the slope of the intraindividual regression is largely independent of the load. The possibility of observer bias was analysed, too. It was found that the standard deviation of the abduction parameters determined by different observers was between 6 and 10% of that between different experimental subjects. RELEVANCE: A knowledge of the kinematics of the shoulder joint is necessary to understand subacromial pathology and in particular impingement problems. The scapulohumeral rhythm depends on the balanced and coordinated function of the muscles involved. The activation pattern of the abductor muscles, as reflected in shoulder kinematics, seems to be individual and stored as an engram. PMID- 11415545 TI - Effects of local muscle fatigue on three-dimensional scapulohumeral rhythm. AB - The relationship of scapulothoracic motion to glenohumeral motion, commonly referred to as the scapulohumeral rhythm, has been the subject of numerous investigations. The purpose of this study was to assess the effects of localized muscular fatigue on three-dimensional scapulothoracic motion and the resulting scapulohumeral rhythm during elevation of the humerus in the plane of the scapula. A six-degree-of-freedom digitizing system (Metrecom(TM)) was used to define scapular and trunk reference frames, and three-dimensional Eulerian angles were determined for scapular motion for 0-135 degrees of humeral elevation before and after fatigue of the upper and lower trapezius muscles. Local muscle fatigue was determined using spectral analysis of electromyographic signals for the upper and lower trapezius muscles. The results demonstrated an average of 22% decrease in the median frequencies of the muscles sampled following resistive exercise, suggesting a state of local muscle fatigue. Concomitant with the fatigue was a selective decrease in scapulothoracic motion about two of the three scapular axes. RELEVANCE: Occupational and recreational conditions often require repetitive overhead elevations of the arms, resulting in muscular fatigue and various pathologies. During repetitive arm elevations the scapulohumeral rhythm or synchronization between the humerus and the scapula is balanced to allow the most efficient elevation of the arm. A better understanding of this much discussed scapulohumeral rhythm and the changes induced by muscular fatigue may provide insight into the pathomechanics associated with shoulder dysfunction. PMID- 11415546 TI - Architecture-based force-velocity models of load-moving skeletal muscles. AB - A predictive model of muscle force-velocity relationships is presented based on functional architectural variables. The parameters of Hill's equation describing muscle force-velocity relationship of nine muscles were estimated by their relationships with variables extracted from the whole-muscle length-force relationship and the percentage of slow-twitch fibres. Specifically, the maximal unloaded velocity (Vo) was estimated through multiple linear regression, from each muscle's fibre composition and the shortening range through which each muscle could produce active force. The maximal isometric force (Po) was also extracted from each muscle's length-force relationship. The ratio of Hill's dynamic constanta to Po and b to Vo, which determines the degree of curvature of the relation, was determined solely by the percent of slow-twitch fibres. This model was verified by fitting it to experimental force-velocity curves of nine different muscles in the cat's hindlimb. It was found that reasonable fits of force-velocity curves would be obtained with correlation coefficient in the range of 0.61 to 0.92, with an average of 0.82. The model predicted that muscles with relatively long shortening ranges would achieve higher maximal velocity, and that muscles with higher percentage of slow-twitch fibres had less pronounced curvature and lower maximal velocity in their force-velocity relationships. RELEVANCE: The results have direct implications in the design of neuroprosthetic limb control systems, which use electrical stimulation to restore function to muscles paralysed from spinal cord injury. The designer is enabled to optimally calibrate the controller according to the predicted individual force-velocity curves of different muscles by using the length-tension curves and fibre composition data available in the literature. PMID- 11415547 TI - A biomechanical investigation of unconstrained shear failure of the osteochondral region under impact loading. AB - This paper presents an investigation into the biomechanics of failure of the osteochondral region under conditions in which the cartilage/bone system was subjected to impact loading in shear through the cartilage layer only. The relationships between the nature of structural integration of the osteochondral region, its mechanism of failure in shear, and its dynamic fracture toughness were examined. The exact mechanism of osteochondral failure was found to be dependent on skeletal maturity. Shear fracture in the immature tissue always occurred subchondrally as a discontinuous propagation of a relatively large, fast moving crack. By contrast, failure in the skeletally mature tissue occurred by the rapid advance of a smaller crack which propagated within the well-defined tidemark region. RELEVANCE: The junction between the compliant articular cartilage and the rigid bone, because it represents an abrupt change in mechanical properties, is a region of potential weakness in the joint system. Clinical findings confirm that failure can occur in the osteochondral region when there is a substantial component of shear loading, and that the immature junction is more susceptible to mechanically induced failure than the mature. This study provides a quantitative assessment of the failure characteristic of the immature and mature osteochondral region under high rates of loading. PMID- 11415548 TI - Effect of maintained stretch on the range of motion of the human ankle joint. AB - The effectiveness of maintained stretch in expanding the range of motion of the human ankle joint was assessed in a population of normal adults. Controlled movements were imposed upon the ankle, and triceps surae and tibialis anterior electromyograms were monitored to ensure that only passive joint properties generated ankle torque. We found that a majority of subjects (7 of 12) showed evidence of muscle activity sufficient to distort a subjective assessment of changes in range of motion. For the remaining five subjects, a 60-s maintained stretch produced a small decrease in the torque subsequently generated by an imposed dorsiflexing movement, but this effect was transient and largely disappeared following 300 s of rest at a neutral position. This short-term effect is consistent with the viscoelastic properties of collagenous material stretched during such treatment and is unlikely to lead to long-term increases in range of motion. RELEVANCE: The results of these experiments indicate that subjective assessments of changes in joint range of motion may be distorted by voluntary and reflexive muscle activation. Moreover the presumed increases in range of motion produced by maintained stretch in our normal subjects were small and transient. These results suggest that future assessments of the long-term efficacy of this treatment must monitor muscle activity and take into account known viscoelastic properties of collagenous materials. PMID- 11415549 TI - Position and orientation in space of bones during movement: anatomical frame definition and determination. AB - This paper deals with methodological problems related to the reconstruction of the position and orientation of the human pelvis and the lower limb bones in space during the execution of locomotion and physical exercises using a stereophotogrammetric system. The intention is to produce a means of quantitative description of joint kinematics and dynamics for both research and application. Anatomical landmarks and bone-embedded anatomical reference systems are defined. A contribution is given to definition of variables and relevant terminology. The concept of anatomical landmark calibration is introduced and relevant experimental approaches presented. The problem of data sharing is also addressed. This material is submitted to the scientific community for consideration as a basis for standardization. RELEVANCE: In order to make movement analysis effective in the solution of clinical problems, a structured conceptual background is needed in addition to standardized definitions and methods. Technical solutions which make data sharing and relevant data banks possible are also of primary importance. This paper makes suggestions in this context. PMID- 11415550 TI - The use of neural networks to recognize patterns of human movement: gait patterns. AB - Artificial neural networks and a statistical method, linear discriminant analysis, were both applied to the recognition of temporal gait parameters associated with altered gait patterns. The duration of the double support and right and left single support phases were measured at seven speeds and three walking conditions. Data from 10 subjects were used to train neural networks, which were then tested using data from 10 other subjects. The overall performance of the networks was at least as high as that of linear discriminant analysis. The relative ease with which neural networks can be set up in a computer, and their discriminatory power, suggests that the technique has a useful role to play in gait analysis. RELEVANCE: The capacity of neural networks to recognize alteration of gait patterns suggests that they might provide an alternative approach for gait assessment. They might be proved to be a useful diagnostic tool. PMID- 11415551 TI - The mechanics of torso flexion: situps and standing dynamic flexion manoeuvres. AB - The mechanics of torso flexion has not been thoroughly investigated due to the lack of knowledge of tissue force-time histories during the performance of torso flexion exercises. An anatomically detailed, three-dimensional model that was sensitive to lumbar curvature and muscle activation patterns was used to assess tissue load distribution during the performance of both isometric and dynamic situps and standing flexion manoeuvres in 12 young men. Situps were performed starting with a flexed torso which was lowered to horizontal and then raised again (instrumentation restrictions did not allow subjects to rest their torsos when in the lowered position). Specific variables measured were air flow during inhalation-exhalation, intra-abdominal pressure, myoelectric activity of the torso muscles, intercostals, and rectus femoris, three-dimensional dynamic curvature of the lumbar spine and body segment displacements. Lumbar compressive loads greater than 3000 N were predicted for both straight leg and bent knee situps. No biologically significant differences were found between bent knee and straight leg situp techniques. RELEVANCE: The safety of situps as both a therapeutic modality and training exercise has been questioned for years but evaluation has suffered from a lack of knowledge of tissue loading. The load predictions in this study suggest that performing situps (both dynamic and quasi static) imposes high levels of compressive loading (> 3000 N) on the low back. The issue of using straight legs or bent knees is probably not as important as the issue of whether or not to prescribe situps at all. PMID- 11415552 TI - A cineradiographic study on the lumbar disc deformation during flexion and extension of the trunk. AB - To reveal deformation behaviour of normal lumbar discs (L((3/4)), L((4/5)) andL(5)/S(1), the lumbar spines of eight asymptomatic volunteers were examined cineradiographically during flexion and extension of the trunk. Disc deformation could be evaluated by displacement of the superior corners of the disc, which were measured with respect to the upper surface of the adjacent lower vertebra. Furthermore the in-vivo strain distribution of each lumbar disc was analysed by the finite element method and in-vivo measurement of the disc deformation. During flexion, deformation of the lumbar disc increased rapidly after a certain delay from the start of trunk motion and reached maximum value before the finish of trunk motion. It was also confirmed that time lags were present between the onsets of disc deformation. Namely, each disc deformed not simultaneously but stepwise from the upper to the lower level with time lags during flexion. When the lower disc deformation started, the strain at the adjacent upper discs had already reached more than half of the value at full flexion. RELEVANCE: The present study revealed one of the phenomena of lumbar spinal kinematics, which will provide helpful information to clinical problems such as evaluation of spinal instability. PMID- 11415553 TI - In-vitro correlation between tension and length change in an anterior cruciate ligament substitute. AB - The length change and tension patterns from multiple insertion locations of an anterior cruciate ligament substitute were studied in 10 cadaver knees. Length change was measured with a spring-loaded isometer of low stiffness, and tension was measured with a piezoelectric load cell. In both instances a thin Kevlar test ligament was positioned in five different femoral and two different tibial ligament insertion locations, that were all located within the normal attachments of the anterior cruciate ligament. Differences were found regarding length changes and tension patterns from a simulated active extension between the central, posterior, and anterior femoral locations. All locations showed larger length change and tension values in extension than in flexion. The anterior femoral ligament insertion location showed length change and tension patterns with increasing values in flexion compared to the other femoral locations. The anterior tibial ligament insertion location showed smaller excursions of both length and tension, than did the central one, but the patterns of the curves were similar. A statistically significant correlation was found between length change and tension patterns throughout a 130-0 degrees range of motion. A statistically significant correlation was also found between the maximum length and tension values. No fixed relationship was found between the magnitude of the length and tension values, when different intervals of the range of motion were studied. RELEVANCE: The intraoperative employment of length change measurements of a test ligament in anterior cruciate ligament reconstruction gives information on where high tension can be expected in the range of motion of the knee, and how this can differ depending on the angle of graft fixation. The information gained can also be used to improve drill channel location. However, no predictions on the magnitude of tension can be made, mainly due to large biological variability. PMID- 11415555 TI - Omnidirectional assessment of one-handed manual strength at three handle heights. PMID- 11415554 TI - Muscular and tibiofemoral joint forces during isokinetic concentric knee extension. AB - The purpose of this study was the examination of muscular and tibiofemoral forces during concentric isokinetic knee extension at angular velocities ranging from 30 to 210 deg s(-1), using a two-dimensional biomechanical model. Five males without knee joint injury history participated in the study. The maximum moment ranged from 226.2 Nm at 30 deg s(-1) to 166.4 Nm at 210 deg s(-1). The maximum muscular force ranged from 7.5 times body weight at 30 deg s(-1) to 5.7 at 210 deg s(-1). The compressive tibiofemoral force ranged from 7.5 times body weight at 30 deg s( 1) to 5.7 at 210 deg s(-1) and the shear tibiofemoral force ranged from 0.9 to 0.8 times body weight respectively. These results indicate that the forces developed during maximal isokinetic knee extension are significantly reduced relative to other powerful dynamic activities but are higher than joint forces during simple walking and cycling activities. Appropriate precautions and adjustment of the isokinetic protocol are required in the final phases of joint injury rehabilitation. RELEVANCE: Isokinetic dynamometry has widespread applications in rehabilitation and training of muscle function. It is therefore important to examine the joint and muscular forces under different conditions in order to alter rehabilitation and training programmes and prevent exercise induced injuries. PMID- 11415556 TI - Surgical correction of leg alignment in unilateral knee osteoarthrosis reduces the load on the hip and knee joint bilaterally. AB - Seventeen patients with strictly unilateral medial knee osteoarthrosis that justified surgical treatment constituted the study group. No patient had any symptoms from the hip or ankle joints or from the other knee. Eight patients were operated with high tibial osteotomy and nine were operated with unicompartmental prosthetic replacement (Brigham model). The forces and the external moment about the hip, knee, and ankle joints of the involved and uninvolved leg were calculated before and 1 year after corrective surgery and compared to the external moment in 10 normal controls. Clinical examinations was performed and the hip-knee-ankle angle in the involved leg was determined from whole-leg weightbearing radiographs before and 1 year after surgery. The external moments about the hip, knee, and ankle joints and the knee moment arms at mid-stance in the frontal plane during gait were determined with a Kistler force plate and a videorecording system. All patients improved clinically after surgery. The mean hip-knee-ankle angle changed from 11 degrees of varus to 0 degrees. In the uninvolved leg the mid-stance adduction moments about the hip and knee joints were significantly increased compared to normal subjects in the control group before surgery, but reduced to a normal level 1 year after surgery. This reduction was mainly due to a significant reduction of the medially directed force. The moments about the ankle joints were not found to be changed by the knee deformity or by the corrective surgery. RELEVANCE: This study demonstrates that a varus malalignment in the osteoarthrotic knee is accompanied by increased load about the uninvolved hip and knee joint. One might expect a more rapid progression of degenerative changes in joint cartilage as a consequence of such increased load. The reduced load after surgical correction of leg alignment might be beneficial to patients with osteoarthrosis of their hip and knee joints. PMID- 11415557 TI - Reliability and validity of the Metrecom Skeletal Analysis System in the assessment of sagittal plane lumbar angles. AB - Seventeen asymptomatic subjects were examined by the Metrecom Skeletal Analysis System and by lateral lumbar radiography. Measurements of sacral base angle, lumbar curve angle, and lumbosacral disc angle were compared with the values derived from the lumbar radiographs. Repeated readings using the Metrecom showed high intraexaminer reliability in measuring these angles. However, the Metrecom values for the lumbar curve and lumbosacral disc angles showed virtually no agreement with those obtained radiographically, while only a weak agreement existed for the sacral base angle. RELEVANCE: Our results suggest that the Metrecom Skeletal Analysis System (version 2.2) is not a valid technique for the determination of either sacral base angle, lumbar curve angle, or lumbosacral angle. It would appear that the values that it generates are more a reflection of back shape than of osseous positioning. PMID- 11415558 TI - Effect of dynamization on gap healing of diaphyseal fractures under external fixation. AB - We asked whether dynamization of externally fixed diaphyseal fractures could improve bone healing in comparison to rigid fixation of fractures having similar remaining gap sizes. To answer this question we evaluated metatarsal osteotomies in 12 sheep. The osteotomy with a 0.6-mm gap was stabilized with a specially designed high bending and torsional stiffness external ring fixator. Osteotomies in six sheep were stabilized rigidly (axial movement < 0.06 mm) or dynamically (axial movement 0.15-0.34 mm). The cyclical axial interfragmentary movement was caused by the load-bearing of the operated limb. With increasing healing time, the initially allowed movement was decreased by callus formation around the osteotomy. The reduction in interfragmentary movement was measured and monitored by a linear variable displacement transducer at the external fixator and a telemetry system. After 9 weeks the sheep were sacrificed and the healed bones were investigated biomechanically and histomorphologically. Compared to the rigidly fixed osteotomies, the dynamized osteotomies showed significantly (P < 0.05) greater (+41%) callus formation and 45% greater tensile strength of the newly formed bone in the cortical osteotomy gap. Histological analysis indicated that the effect of dynamization occurred mainly after the 5th week. RELEVANCE: From these results we conclude that dynamic fixation of diaphyseal gaps is advantageous in comparison to stable external fixation. PMID- 11415559 TI - Peripheral nerve tension due to joint motion. A comparison between embalmed and unembalmed human bodies. AB - Various joint positions of the upper extremity were used to study the tensile forces on the median nerve. To analyse the effect of embalmment, tensile forces were measured in situ in unembalmed and embalmed human bodies. A positive correlation was found between tensile force data from unembalmed and embalmed nerves. This finding justifies, for comparative studies, the use of embalmed human bodies, although the absolute tensile forces are higher. RELEVANCE: In daily activities or during certain operations positions of the upper extremity vary, hence creating tensile forces of different magnitude on peripheral nerves. Tensile forces are also applied in the diagnosis of nerve (root) lesions of the upper extremity. To analyse these tensile forces, in-situ experiments on unembalmed human bodies, though problematic, are supposed to be the most realistic approach. In this study it has been shown that in comparative studies on peripheral nerve tension, data obtained from embalmed human bodies can be used. PMID- 11415560 TI - Mechanical tension in the median nerve. The effects of joint positions. AB - Stretch tests are attractive in the diagnosis of nerve root or peripheral nerve lesion. Interpretation of the test results is often difficult since the distribution of tensile forces along the nerve caused by the test manoeuvre is not known. In this study the effect on median nerve tension of 22 positions of the arm was measured with 'buckle' force transducers. With the elbow in full extension and the hand in neutral position, altering the position of the shoulder significantly influenced tension in the proximal part of the median nerve; tension in the distal part was not influenced. With the shoulder in 90 degrees abduction, dorsiflexion of the hand combined with an extended elbow resulted in an increased tension in both distal and proximal parts of the median nerve. Dorsiflexion of the hand combined with flexion of the elbow caused an increase in tension only in the distal part. At all sites of the median nerve the median nerve upper limb tension test caused a significantly higher tension than the radial and ulnar nerve upper limb tension tests. This study provides insight in the normal distribution of tensile forces along the median nerve and can have clinical consequences. For differentiating nerve root from peripheral nerve lesions a specific provocative tension test for the median nerve is advocated. The results of this study provide a theoretical basis for differentiating between lesions in the proximal and distal parts of the median nerve. RELEVANCE: In the diagnosis of nerve(root) lesions, tests in which stretching the nerve provokes the symptoms are widely used, but no data are available on the distribution of tensile forces along nerves and nerve roots during such tests. In this study a human cadaver model is presented to analyse tensile force distribution on the median nerve; it can be an aid to evaluate the specificity of a clinical provocation test for the median nerve. PMID- 11415561 TI - In vitro method for quantifying the effectiveness of the longitudinal arch support mechanism of a foot orthosis. AB - The purpose of this investigation was to develop a technique to quantify the effectiveness of the longitudinal arch support mechanism of a foot orthosis. The experimental model was based on the following principle of foot biomechanics: as the foot is subjected to a load, a proportion of the load is experienced as tension by the plantar aponeurosis. A differential variable reluctance transducer was implanted into the plantar aponeuroses of cadaveric lower limb feet through which the strain was calculated in three conditions, specimen barefoot, specimen with shoe, and specimen with shoe and orthosis. Each donor limb was mounted in an electromechanical test machine that applied a load of 900 N to the tibia. Time, load, and strain data were collected and analysed at four load levels (225, 450, 675, 900 N). In addition the measurements and test design were evaluated for reliability. Strain in the plantar aponeurosis decreased significantly in the specimen with shoe and orthosis compared to the specimen with shoe only. There was a significant increase in the time to load data in the specimen with shoe and orthosis condition in contrast to the barefoot measurements. There were no significant differences in strain between the barefoot tests and those of the shoe, indicating that the shoe tested provided minimal support to the foot's longitudinal arch. RELEVANCE: One of the most common foot pathologies that patients seek medical attention for is plantar fasciitis. The primary cause of this condition is excessive tension in the plantar aponeurosis. A foot orthosis is often prescribed for treatment, relying on its longitudinal arch support mechanism to relieve the strain in the plantar aponeurosis. Quantifying the amount of strain experienced by the plantar aponeurosis is needed to identify how effective foot orthoses are in providing support to the foot's longitudinal arches. Such information is of importance to the medical practitioner who is involved with orthotic clinical recommendations. The described method will also be useful to bioengineers concerned with the arch support component of running shoes. PMID- 11415562 TI - Influences of inversion/eversion of the foot upon impact loading during locomotion. AB - Pronation of the foot is believed to be one of the mechanisms used during locomotion to attenuate the loading experienced by the body at ground contact. The purpose of this study was to quantify the changes in loading induced by modifications to the normal pronation of the foot during walking and running. Impact loading in 10 subjects was determined using ground reaction force and tibial acceleration. The results indicated that impact loading was increased when normal pronation was prevented during running. However, there was no reduction in impact loading when normal pronation was exaggerated. RELEVANCE: Orthotic corrections are commonly prescribed to control excessive foot pronation in patients who experience knee pain. The findings of the present paper demonstrate that pronation modifications can affect the magnitude of the impact experienced by the body during locomotion. It was also found that the loading responses to pronation modifications were not consistent between walking and running. Thus care should be exercised when orthotics that will be used for both walking and running are prescribed. Furthermore, patients should be informed that their orthotics may be activity specific. PMID- 11415563 TI - Viscoplasticity of rabbit skeletal muscle under dynamic cyclic loading. AB - To elucidate the strain effects of dynamic cyclic loading on the skeletal muscle tendon unit, the muscle-tendon unit of New Zealand white rabbits were tested with different strains and strain rates under ketamine general anaesthesia. The results indicated that the dynamic responses of muscular tissues were both strain and strain-rate dependent with appreciably non-linear characteristic. The muscle tendon unit behaved viscoelastically below 12% strain; but when stretching more than 16% strain it began to behave plastically. The loading energies and unloading energy are more dependent on strain than on strainrate. RELEVANCE: This study offers firm evidence that plasticity may be one of the important biomechanical properties of the skeletal muscle. The findings in this report imply that the expression of skeletal muscle mechanical behaviour may not be limited to phenomena represented by a composition of elastic and viscous elements. The elongation of skeletal muscle under sustained stretching might be explained by the plastic property. Elucidation of the existence of viscoplasticity in skeletal muscle gives a good reference point to determine the functional capabilities and limitation of muscular tissue. PMID- 11415564 TI - Biomechanical properties of the triceps surae muscle after limb lengthening. AB - The hindlimb of New Zealand white rabbit was osteotomized and then slowly lengthened at a rate of 1 mm/day until a 2.0-cm gain in length was reached. The triceps surae muscle-tendon unit was then tested either with or without 2 months fixation for bony consolidation. After limb lengthening, the strain at peak load was decreased and the axial rigidity was increased significantly; but other biomechanical parameters such as peak load, maximal deformation at peak load, stiffness, and energy absorption before peak load did not show any significant difference. We conclude that healthy triceps surae muscle has a great potential to be lengthened despite the changes in intrinsic structural properties of the muscle. The change in the biomechanical properties occurred during the time of distraction and was not affected by the time of bony consolidation. RELEVANCE: As noted in this study, many biomechanical parameters of the triceps surae muscle were not affected after prolonged distraction. This fact suggested that the healthy muscle-tendon unit is not a main causative factor of joint contracture after limb lengthening. However, there were intrinsic changes in the structural properties of the triceps surae muscle. Equinus contracture after limb lengthening may be caused by the aggravated intrinsic structural changes of the injured or denervated scarred muscular tissue. PMID- 11415565 TI - Formation of impacted fractures of the femoral neck: an experimental study. AB - Nine femoral-neck specimens were exposed to a fracturing force applied to the femoral head, perpendicular to the axis of the femur, simulating a fall onto the greater trochanter. The mechanical strength and fracture type were investigated in a universal testing machine. In all nine specimens, the superior (cranial) cortex of the femoral neck fractured first, followed by some compression and fracture of the inferior cortex, whereby impacted fractures had been formed. After application of additional force, dislocated fractures occurred. This method may be of value in experimental studies of impacted fractures of the femoral neck. RELEVANCE: This study describes an experimental procedure for formation of impacted femoral neck fractures. Hopefully this technique may be employed in further examining the nature and biomechanics of impacted hip fractures. The ultimate aim would seem to be a classification with regard to which fractures require surgery and which may be treated conservatively. PMID- 11415566 TI - Influence of sensory input on plantar pressure distribution. AB - The sensory feedback control system plays a central role in human locomotion. However, few studies have been published discussing the influence of sensory input at the plantar surface of the foot on the kinematics and kinetics of locomotion. The purpose of this study was to examine the effect of changes in sensory input at the plantar surface of the foot on the pressure distribution measured at the plantar surface of the foot. Three pairs of specially made socks with sand at the plantar surface of the foot and one pair of normal socks were used to simulate different sensory inputs. Pressure distribution at the plantar surface of the foot was measured using an emed system for 10 subjects for each of the four sensory input conditions during walking and running on a treadmill. The results showed significant and substantial changes in pressure in selected areas for changes in sensory input. The pressure increased in the midfoot area and decreased in the toe area with increasing sensory inputs. These results are similar to results of a previous study where similar changes were found in a comparison between comfortable and uncomfortable shoes. RELEVANCE: Pressure distribution is increasingly used in clinical assessments. However, the factors influencing the pressure distribution pattern are only partially understood. One factor may be the sensory input into the foot. It is important to understand this influence in order to provide appropriate interpretation of the pressure distribution data. PMID- 11415568 TI - Ankle laxity testing device for in vivo use-- PMID- 11415567 TI - Flexible electrogoniometers: kinesiological advantages with respect to potentiometric goniometers. AB - Unlike conventional potentiometric goniometers, flexible electrogoniometers do not suffer from any alignment problems with respect to the joint axis. We hypothesized that flexible goniometers provide more valid measurements in that they avoid some biomechanical pitfalls. With both devices simultaneously we measured the movements of dorsal flexion of the ankle and flexion of the knee in three healthy subjects. In the various subjects, this comparison showed that the flexible goniometers signalled ankle excursion greater by 19-40% with the foot remaining in neutral position or being simultaneously pronated, and lower by 10 21% with the foot being simultaneously supinated. At the knee the flexible goniometers signalled a flexion greater by 24-32%, with respect to potentiometric goniometers. Biomechanical considerations support the validity of records taken with flexible goniometers, potentiometric measures being biased by (a) multijoint, multiplane motions underlying foot dorsal flexion, and (b) multiaxis motion underlying knee flexion RELEVANCE: In measurements of angles between adjacent body segments, flexible goniometers should be preferred to potentiometric goniometers despite their higher cost. Flexible goniometers not only are more practical: more importantly, they provide a valid measure of relative orientations in one plane, regardless of the number and different concurrent motions of the underlying joints. PMID- 11415569 TI - Measurement, nature, and implications of skeletal muscle strength in patients with neurological disorders. AB - Muscle strength is frequently impaired in patients with neurological disorders. Numerous instrumented and non-instrumented options exist for measuring the strength of such patients. Such measurements are useful for clarifying patient status and documenting changes over time. Moreover the measurements are often informative of present or future function among a variety of diagnostic groups. Measurements of muscle strength are an essential component of the neurological evaluation and provide information of substantial importance to clinical care. PMID- 11415570 TI - Clinical evaluation of a system for precision enhancement in spine surgery. AB - Most techniques in segmental spinal fixation surgery rely on the identification of predefined targets with the help of anatomical landmarks and on intraoperative use of image intensifiers. However, because there is no direct link between the image information, the accessible spinal anatomy, and the action of surgical instruments several potential problems and possible complications are still involved. A novel system for spinal surgery has been designed allowing for the real-time, intraoperative localization of surgical instruments in medical images. In practice this was achieved by combining image-guided stereotaxis with advanced optoelectronic position sensing techniques. Modules were developed for image data processing, surgical planning and simulation, and various intraoperative procedures. A detailed validation of the system was performed indicating an overall accuracy to be better than the slice distance of the spinal image used. In an in-vitro setting 20 pilot holes for pedicle screws were prepared in human cadaveric lumbar spines. An analysis in 77 histological cuts showed an ideal location in 70 and only minor cortex engagement in seven sections. In vivo the system has been successfully applied in three posterior low lumbar stabilizations with overall 15 transpedicular screws. RELEVANCE--:This article focuses on the clinical evaluation of a computer-assisted surgery system and its application to the operating theatre for transpedicular fixation of the spine. The given approach effectively keeps the surgeon 'in the loop' and requires only minor modifications of the established surgical techniques and associated instruments. The results of this study indicate that advanced computer-assisted techniques may significantly improve the accuracy and safety of surgical interventions of the spine. The proposed technique may in future be adapted to other applications in orthopaedic surgery. PMID- 11415571 TI - Biomechanical study on femoral neck fracture fixation in relation to bone mineral density. AB - Twenty pairs of fresh-frozen cadaveric femurs were used in the study. The left femurs were used as control for the mechanical testings and bone mineral content scans, the right femurs were divided into two experimental groups, i.e. 'fractured' group and 'healed' group. In the 'fractured' group, twelve right femurs were osteotomized at the plane perpendicular to the femoral neck shaft axis, through the mid-cervical area of the femoral neck. The artificially created fractures were fixed with the AO dynamic hip screw system using a 4-hole plate and a compression screw. In the 'healed' group, the dynamic hip screws were applied to eight intact right femurs to simulate healed fractures. Bone mineral density scans and mechanical testings were performed on all the femurs. Good correlation was observed between bone mineral density and femoral neck strength in the control group. There was a decrease of 43.5% in strength in the 'fractured' group when compared to the control group. However, in the 'healed' group the failure load was found to be 15.2% lower than the control group. The femoral fixation strength in the 'fractured' and 'healed' groups had good correlation with the bone mineral density. RELEVANCE--:Results from this study indicated that bone mineral density is an important predictive factor in fracture fixation failure. Therefore it may be appropriate to consider the bone mineral density of a patient with proximal femoral fracture treated with fixation devices, as a criterion in prescribing a more protective postoperative management, with respect to weight bearing protocol. PMID- 11415572 TI - Muscular synergy in the shoulder during a fatiguing static contraction. AB - The synergic operation of shoulder muscles during a fatiguing submaximal arm flexion task was studied using both a biomechanical 3-dimensional shoulder model and electromyographic recordings. A new optimization scheme aiming to maximize the task endurance time by constantly regulating the force output of each muscle is utilized in the model. The method can be used to simulate the muscle rotation phenomenon, which has been proposed to occur during an endurance type contraction. The model predictions on the fatigue order of the shoulder muscles were compared to results derived using the median frequencies of the electromyographic signals from nine muscles or muscle parts. In the test performed 10 men held until exhaustion a weight (4 kg) suspended on the wrist with the arm in horizontal flexion. The deltoid, infraspinatus, and supraspinatus muscles were the first to show electromyographic signs of fatigue. The times for detecting electromyographic changes in the trapezius muscle were longer than those in the muscles first showing electromyographic signs of fatigue. The biomechanical model used predicted the upper and lower infraspinatus and the anterior part of the deltoid to be the first to show signs of fatigue during the flexion task. The predictions of the biomechanical model on the order of fatigue of the nine shoulder muscles monitored also using electromyographic recordings corresponded to the electromyographic results. However, the accuracy of this comparison is limited by the fact that the electromyographic recordings did not cover all the muscles used in the model. No clear order was found for the development of electromyographic signs of muscle fatigue. RELEVANCE--:The biomechanical model and the developed optimization methods provide new tools for studying the synergic operation of shoulder muscles during fatiguing contractions. Our results will help in analysing and optimizing shoulder load in problematic functions both of work and leisure activities. PMID- 11415573 TI - Postural responses to platform perturbation: kinematics and electromyography. AB - Dynamic postural control may be studied using feedback paradigms which disturb standing balance. In this study the kinematics and electromyography of the reaction to an unexpected slip were examined simultaneously in 16 young, healthy volunteers to characterize the response to postural perturbation in the context of the perturbation characteristics. Kinematically, results showed ankle plantarflexion, knee flexion, hip extension, and shoulder flexion. Electromyographically, sequences of activation of the postural muscles of the leg were primarily anterior distal. Data is presented that supports a new model of the reaction to perturbation which incorporates the timing and type of muscular activity, the type of motion (active or passive), and the perturbation characteristics. RELEVANCE--:Understanding the interplay of the neurophysiological and biomechanical aspects of dynamic postural control gives insight into balance reactions. Simultaneous consideration of the kinematics, muscle activation, and perturbation characteristics allows a more comprehensive view of recovery of balance. PMID- 11415574 TI - Static back endurance and the risk of low-back pain. AB - Spinal physical capacity and in particular static back endurance were studied for their prediction of first-time experience of low-back pain. Of a total of 126 persons who were free from back complaints at entry, 33 developed low-back pain during a follow-up of 1 year. The static back endurance test was found to be the only physical capacity measurement that indicated an increased risk of low-back pain. Adjusted for age, sex, and occupation, the odds ratio of a new low-back pain in those with poor performance was 3.4 (95% confidence interval, 1.2-10.0) compared to those with medium or good performance. RELEVANCE--:The simple static back endurance test might have value in health examinations because of its strong predictive value of future low-back pain. PMID- 11415575 TI - Correlations of isokinetic and psychophysical back lift and static back extensor endurance tests in men. AB - Isokinetic lift, psychophysical lift, and static back endurance tests are among the most widely used measurements of muscle function for determining risk for, and recovery from, back problems. After determining test repeatability we examined the degree of association between these test measures in 100 men selected to represent a variety of occupations and lifestyles. Isokinetic lifting had low correlations with test results for psychophysical lifting (r = 0.28) and static back endurance (r = 0.24). Static back endurance and psychophysical lift test results were not at all correlated (r = 0.03). RELEVANCE--:Muscle function measurements for back problems are sometimes arbitrarily selected due to health practioners' beliefs and preferences or practical reasons such as the availability of equipment. Yet little is known about the associations of these tests with one another or with anthropometric factors. The data presented in this study document that isokinetic lift, psychophysical lift, and static back endurance tests appear to measure very different aspects of muscle function, and comparing studies using these different test methods should be approached with these low inter-test correlations in mind. PMID- 11415576 TI - Response of immature diabetic rat bone-ligament junctions to insulin and exercise. AB - The mechanical and morphological characteristics of femur-medial collateral ligament-tibia units and the histomorphometry of medial collateral ligament tibial insertion were examined in female Sprague-Dawley rats with diabetes mellitus (type I, insulin-dependent diabetes). Diabetes was induced with the streptozotocin, a drug with toxic effects on insulin-producing islet cells in the pancreas. The groups studied included rats with untreated streptozotocin-induced diabetes mellitus (Diabetes), insulin-treated streptozotocin-induced diabetes (Diabetes-Insulin), exercise with streptozotocin-induced diabetes (Diabetes Exercise), exercise with insulin-treated streptozotocin-induced diabetes (Diabetes-Insulin-Exercise), and age-matched sedentary-control rats (Control). Diabetes and Diabetes-Exercise groups had significantly lower body mass and higher blood glucose than Diabetes-Insulin, Diabetes-Insulin-Exercise, and Control groups, indicating that exercise alone did not prevent growth retardation or improve blood glucose control in the streptozotocin-induced diabetes. The strength of Diabetes femur-medial collateral ligament-tibia units was significantly less than Control, but exercise (with or without insulin treatment) maintained the bone-ligament-bone unit strength at a normal level. The load (per unit body mass) of Diabetes-Exercise femur-medial collateral ligament-tibia unit was significantly greater than Control, Diabetes, and Diabetes-Insulin groups. The tensile stiffness (per unit body mass) of the femur-medial collateral ligament-tibia unit for the Diabetes-Exercise group was also significantly greater than Control and Diabetes-Insulin groups. The fibroblast-like cell density in medial collateral ligament at its tibial-insertion site and medial collateral ligament insertion area was significantly less in Diabetes rats. RELEVANCE--:Diabetes mellitus can have deleterious effects on connective tissues, such as bone and ligament. Thus the bone-ligament junction becomes vulnerable. The results of this experiment suggest, however, that exercise, with or without insulin treatment, can have a positive effect on the bone-ligament interface in the rat with type I diabetes mellitus. Furthermore exercise (with insulin treatments) can mitigate growth retardation and many of the adverse effects of streptozotocin-induced diabetes on the structure and mechanical properties of immature bone-ligament junctions. PMID- 11415577 TI - Mechanical properties of the human anterior cruciate ligament. AB - The aim was to measure the stiffness and strength of the femur-anterior cruciate ligament-tibia complex tested in a physiological manner with a force exerted anteriorly on the tibia, at knee joint flexion angles of 0 degrees, 10 degrees and 30 degrees and at speeds of 50 and 500 mm/min. Ligaments were preconditioned by cycling five times, with data from the fifth cycle used to determine the stiffness of the ligament in a low-load range. The ligaments were then tested to failure with the knee at 30 degrees flexion. The specimens were divided into two groups, middle-aged (40-60) and old (>60). For each group no statistical difference was observed between stiffness of the ligament at different joint flexion angles or speeds. Seven of the 21 specimens in the older age group failed by avulsion at the bone-ligament interface. All the other specimens failed by tears in the substance of the ligament. Ultimate failure load was found to have a significant correlation with bodyweight. It was 1.6 and 1.3 times bodyweight for the middle-aged and older age groups respectively. This study has highlighted the importance of identifying different modes of failure, of making corrections for bodyweight and testing in a physiological manner. The results allow a better understanding of the mechanical behaviour of the anterior cruciate ligament and provide design data for anterior cruciate ligament grafts and prostheses. RELEVANCE:--Our clinical experience indicates that the anterior cruciate ligament is frequently ruptured during uncoordinated contraction of the quadriceps mechanism. The results of this study, in which the mechanical properties of the anterior cruciate ligament have been measured with force exerted anteriorly on the tibia, allow a more complete understanding of the mechanical behaviour of the anterior cruciate ligament and provide design data for anterior cruciate ligament grafts and prostheses. PMID- 11415578 TI - Long-term effects of hindfoot fractures evaluated by means of plantar pressure analyses. AB - The present study investigated the long-term effects of calcaneal fractures on the plantar pressure distribution during human gait. Fourteen patients participated in a follow-up study and were assigned to a group with good or poor calceneus restoration as indicated by normal or reduced Bohler's angle. Ten normal subjects with similar anthropometric characteristics were used for comparison. Gait analysis was performed by means of a capacitive pressure distribution platform. A comparison of the injured with the healthy foot of all patients revealed a statistically significant peak pressure decrease under medial areas of the foot (first metatarsal -24%) and an increase in lateral areas (midfoot +135%, fifth metatarsal +122%). This load shift towards a lateral loading indicates a persisting deviation of the gait pattern long after healing. Furthermore we investigated whether the success of the restoration of hindfoot geometry as reflected in Bohler's angle had an effect on the foot loading characteristics. Therefore we compared the gait patterns of the two patient groups with the normal subjects. However, the results did not reveal more abnormal pressure patterns in the poorly reduced group. RELEVANCE:--The restoration of the shape of the calcaneus with joint surface congruity and reconstruction of Bohler's angle appears generally accepted as a major aim in the treatment of calcaneal fractures. While this goal can usually be achieved by contemporary surgical techniques, the restoration of foot function under dynamic loading conditions is not guaranteed and should be assessed objectively. The investigation of the foot loading characteristics by means of plantar pressure measurements can be used to support the evaluation of the patient's recovery after trauma. PMID- 11415579 TI - The mobility of the sacroiliac joints in healthy volunteers between 20 and 50 years of age. AB - The nature and amplitude of movement in the sacroiliac joint (SIJ) is still open to controversy. Whereas some authors using modern measuring techniques have reported on the range of motion in the SIJ of patients and in embalmed elderly humans, the following is a presentation of our observations related to healthy individuals between 20 and 50 years of age. Using a three-dimensional stereophotogrammetric method, the motion in the joints of 15 males and nine females was investigated with change in posture from the upright standing position. The general description of spatial motion, as obtained through the helical axis concept, has been used. For comparison of the results obtained, the motion is also specified as components of rotation about vertical, anteroposterior and transverse axes, or in horizontal, frontal and sagittal planes respectively. The average values for total rotation and translation were low, being 1.7 degrees and 0.7 mm respectively. One of the test subjects who was known to have occasional trouble with his sacroiliac joints exhibited more than 6 degrees rotation. No statistically significant differences could be demonstrated with respect to sex, age, or parturition. RELEVANCE:--No data are available in the literature on the motion of the sacroiliac joints of healthy men and women in the age group 20-50 years. Measurements were carried out with the aid of percutaneously introduced external markers, using conventional light photography. This is a definite advantage over the use of X-rays and radio-opaque markers that would probably remain implanted in the bone indefinitely. Therefore this method might also be considered for future use in clinical research involving the mobility of the SIJ in patients. PMID- 11415580 TI - Extension creep in the lumbar spine. AB - This in vitro study was designed to examine extension creep of the lumbar spine under conditions of sustained loading. By means of a weight and pulley circuit, loads simulating lumbar extension and flexion were applied to 25 male lumbar spines removed at postmortem. Sagittal ranges of movement and the amount of creep occurring in a 20-min period at the limit of extension were recorded. The data were classified into three age group categories representing young adults, the middle-aged and the elderly. The results indicated that extension creep behaviour was similar to that of flexion creep previously reported. The elderly lumbar spines appeared to have a greater range of extension creep than those of the young and middle-aged groups, but analysis of variance showed that the difference in creep between age groups was not significant. The viscoelastic behaviour of the spinal ligaments, zygapophyseal joint capsules and intervertebral discs during extension are sufficient to produce a characteristic creep-recovery curve, but perhaps the important role that the zygapophyseal joints play in limiting extension is sufficient to prevent significant change in creep behaviour with increasing age. RELEVANCE:--This study provides information on the biomechanical responses of the lumbar spine to sustained end range extension loading. Results suggest that in the living, even brief periods of sustained extension loading may cause significant extension creep. PMID- 11415581 TI - Isokinetic strength relationships in shoulder muscles. AB - The purpose of this study was to extend and refine former findings which related to strength characteristics of the major muscle groups operating on the glenohumeral joint. Based on preliminary isokinetic evaluation of five women and five men, which indicated that overall dominance was not associated with significantly stronger muscles, the dominant shoulders of 30 normal non-athletic subjects, 15 women and 15 men, were tested concentrically and eccentrically, at 60, 120 and 180 degrees /s. The descending order of muscle strength at both contraction modes and for both genders was: extensors, adductors, flexors, abductors, internal rotators and external rotators. Inter-muscle groups' strength correlation coefficients have indicated significant, moderate-fairly strong, relationships in men but not in women. The implications of these findings in terms of testing and shoulder muscle conditioning are discussed. RELEVANCE:- Analysis and interpretation of shoulder muscle dysfunction, and implementation of conditioning protocols, require the availability of normative values pertaining to the common modes of contraction and relative to an acceptable and velocity setup. This study provides a representative data base of normal subjects which may serve for these purposes. PMID- 11415582 TI - In vivo effect of shock-waves on the healing of fractured bone. AB - In a controlled animal experiment we attempted to clarify the question of whether there is a stimulating effect of extracorporeal shock-waves on the repair process of fractured long bones. As a fracture model we used an osteotomy in the diaphysis of the ovine tibia and an external fixation device. Shock-wave treatment at two levels of intensity and with four different numbers of applied shocks was performed with an electromagnetic acoustic source. Healing of the osteotomized bone was evaluated by biomechanical and radiological investigations on the whole bone as well as on bone sections from areas of the fracture gap and the periosteal fracture callus. We found a non-significant tendency to deterioration of the fracture healing with increasing shock-wave intensities. The study of treatment parameters led neither to significantly different biomechanical outcomes nor to altered radiological results in comparison to the untreated control group. RELEVANCE:--While we cannot comment upon the effectiveness of extracorporeal shock-waves in the delayed treatment of fractures or pseudarthrosis, our results suggest that shock-waves have no beneficial effect in acute fracture repair. PMID- 11415583 TI - Effects of various isoresistive training programmes on trunk muscle performance. AB - Isoresistive movements provide a functional method of testing muscle performance and of providing exercise; the resistance applied is usually a percentage of the maximal isometric torque which can be developed by individual subjects. The aim of the current study was to compare the effectiveness of dynamic isoresistive exercise programmes using 25, 50 or 75% of the maximum isometric flexion torque and static isometric exercises for improving the performance of the trunk flexors and extensors. Subjects comprised 42 asymptomatic females aged 18-25 years who were randomly assigned to control or exercise groups. The exercise groups undertook 2 min of exercise 3 times per week. The control subjects showed significant improvement between tests indicating a strong learning effect. The study showed that the subjects who exercised at 50% of maximum flexion isometric torque achieved the greatest increases in muscle performance on most measures. maximal isometric flexion and extension torques did not significantly change in any of the groups. Flexion and extension velocity and power were the muscle performance characteristics which improved the most in response to the training programmes. RELEVANCE:--This paper attempts to determine which among several options is the most effective isoresistive protocol for conditioning trunk muscle performance, as improved strength and power of these muscles may contribute in preventing episodes of LBP. PMID- 11415584 TI - Development of a connectionist expert system to identify foot problems based on under-foot pressure patterns. AB - The first phase of developing a connectionist expert system is described in this study. Dynamic pressure patterns below the foot obtained by the emed f system of 18 subjects were classified into one of the three categories: normals, pes cavus and hallux valgus. As opposed to the traditional approach for the analysis of the pressure patterns by quantifying isolated segments, a neural network based expert system was trained to perform objective classification of the patterns while keeping their integrity. The results of the backpropagation paradigm were unexpectedly good considering the limiting factors which hindered the performance of the neural networks. The successes indicate the potential of artificial intelligence in automated analysis of foot pathologies based on theemed f system. RELEVANCE:--This paper describes the first steps towards a novel automated screening system, which classifies the patient with foot abnormalities into the pathological category he or she belongs to, based on the pressure pattern below the foot. The decision making is objective, and shows a practical example of the use of artificial intelligence. PMID- 11415585 TI - Fracture stiffness measurement in tibial shaft fractures: a non-invasive method. AB - This paper presents a non-invasive method of assessing healing by measurement of fracture stiffness. The method works on the principle that if the load (F) applied at a certain known distance (Y) from the fracture is measured, then the moment (M = FY) at the fracture site can be calculated. By measuring the angle/deflection (straight theta) occurring at the fracture site using a suitable instrument (electrogoniometer), the necessary data to calculate fracture stiffness (FY/straight theta) would be available. The method was employed to assess the stiffness in a series of tibial shaft fractures treated conservatively, all of which healed uneventfully. This paper concentrates on a group of tibial shaft fractures in which the radiological criteria for fracture union were not satisfied even after a mean duration of 20 weeks treatment. The non-invasive method of measuring fracture stiffness supported the clinical impression of union in most cases at the first test, but was repeated on two more occasions to confirm the trend of progressive healing. The objective evaluation of fracture healing led to avoidance of surgical intervention in these patients, who went on to sound union. PMID- 11415586 TI - Comparison of the mechanical performance of three types of external fixators: linear, circular and hybrid. AB - Different configurations of the Monticelli-Spinelli and Ilizarov external fixation systems were tested to define their mechanical properties. In five configurations the external fixator consisted of rings with tensioned wires (circular), while in one configuration two pairs of the tensioned wires and their correspondent ring were replaced by threaded pins (hybrid). Testing was performed in axial compression, bending and torsion. The results were compared to the characteristics of a selected linear fixator group. Both the circular and the hybrid configurations were non-linear in compression. In bending, circular fixators had a similar pattern in both anteroposterior and oblique loading directions. The bending load-displacement pattern for the hybrid fixators was similar to the linear fixators, higher stiffness in the plane of the pins. Torsion was linear for both circular and hybrid fixators, as for the linear fixators. By combination of wires and pins (hybrid configuration), the mechanical behaviour had characteristics from both linear and circular fixators. It is concluded that the three studied groups own different mechanical performance and can be considered as different types of fixators. While it has been demonstrated that osteogenesis can be achieved independently of the mechanical behaviour of the fixator, this study supports the suggestion that some complications can be related to the mechanical behaviour of the fixator. PMID- 11415587 TI - Dynamic, semidynamic and static analyses of heavy lifting: three approaches applied to sack handling. AB - The load on the lower back was calculated by means of biomechanical models of different degrees of complexity for twelve subjects lifting 30-kg sacks from three different heights. Motions and ground force reactions were registered by means of a video camera, an advanced optoelectronic measuring system, and a force plate. The results showed that static analyses underestimated the load on the back. However, fully dynamic analyses were not required but could be replaced by simpler 'semidynamic' analyses. The experiments were carried out during two periods with an interval of 5 months. The calculated loads on the back were larger during the first period. This could possibly be explained by differences in the experimental setup or by the fact that the subjects were very young, 16-17 years, and may have increased in growth and strength during the interval. PMID- 11415588 TI - Predictive model of intersegmental mobility of lumbar spine in the sagittal plane from skin markers. AB - This article describes a sagittal motion model that transforms skin marker coordinates into corresponding vertebral body coordinates. Coordinate data were obtained from 45 subjects with radiopaque skin markers over the vertebral spinous processes. Data of 30 subjects were used for model development while data of the other Fifteen subjects were used for model validation. The subjects were subjected to lateral spinal radiographs in positions of neutral, flexion, and extension. The model was developed by taking the non-linear difference between lumbar spinal profile and skin profile, the L(4) skin-fold, and the difference of L(1)-S(1) skin distraction into consideration. Regression analysis shows high coefficients of determination (range, 62.43-99.86%; mean, 86.41%) in comparing the estimated position data with the actual data in all vertebral body positions. Based on the estimated coordinates, both the intersegmental joint angles and intersegmental joint mobilities were calculated. The result of this study using the non-invasive protocol and the transformation matrices demonstrated that the intersegmental sagittal motion of lumbar spine can be obtained with error ranged from 0.05 to 0.56 degrees. PMID- 11415589 TI - Effect of torso position on arthrometric assessment of anterior knee laxity. AB - The purpose of the study was to examine the effect of changes in torso position on arthrometric assessment of knee laxity. Anterior tibial translation (ATT) of 12 control and 12 ACL deficient subjects was assessed using a Dynamic Cruciate Tester during five maximal isometric knee extensions and five passive tests (240 N load) on the right and left knee in each of three torso positions: vertical, reclined, and supine. Electromyographic activity of rectus femoris, vastus lateralis, biceps femoris, and semimembranosus were sampled during arthrometric assessment. Results indicated there was no significant difference in mean ATT as a function of torso position. However, subjects demonstrated significantly greater hamstring activity when seated vertically compared to reclined or supine and significantly greater hamstring activity when reclined compared to supine. PMID- 11415590 TI - Effect of elongation rate on the failure properties of the rabbit anterior cruciate ligament. AB - The effect of elongation rate on the failure properties of the rabbit femur anterior crucicate ligament-tibia were investigated. Paired limbs were elongated to failure at rates of 0.0001 m s(-1) and 0.92 m s(-1). Two distinct types of tibial avulsion injury reflecting rate-dependent areas of weakness were noted. 'Bony avulsions' formed by the junction between cortical and trabecular bone comprised the predominant injury observed at the faster elongation rate. 'Fibrous avulsions' between the zones of mineralized fibrocartilage and bone occurred predominantly at the slower elongation rate. The faster rate significantly increased ultimate load (74%) and stiffness (615%) on average, relative to the slow rate. In contrast to what has been previously described, there was a significant decrease in failure deformation (79%) at the faster rate. PMID- 11415591 TI - The potential of magnetic resonance imaging (MRI) for quantifying articular cartilage thickness -- a methodological study. AB - The thickness of patellar articular cartilage was assessed in a cadaveric human knee joint by magnetic resonance imaging. Imaging was conducted at 1.0 T, using three-dimensional gradient-echo sequences. From each of the sequences the total cartilage volume, the size of the articular surface, the mean cartilage thickness and the regional distribution of cartilage thickness were determined by image analysis. These values were then compared with those obtained from anatomical sections. The fat-suppressed FLASH sequence was found to allow the most accurate evaluation of the total volume and the regional distribution of the articular cartilage. Slight underestimation of the cartilage thickness by about 5% may be due to the fact that the calcified layer is not made visible by magnetic resonance imaging. There is, however, a very high degree of similarity between the distribution patterns obtained from the MR images and the anatomical sections. The contrast-to-noise ratios and reproducibility were also highest with the fat-suppressedFLASH sequence. This pulse sequence can therefore be recommended for experimental and clinical use. PMID- 11415592 TI - Effect of directional symmetry on the torsion testing of paired long bones. AB - Paired long bones (canine humeri) demonstrated no significant biomechanical difference between left and right limbs torsioned tested symmetrically (a) both internal: (b) both external or asymmetrically, (c) right internal/left external. PMID- 11415593 TI - Mechanical stability of the in vivo lumbar spine: implications for injury and chronic low back pain. AB - One important mechanical function of the lumbar spine is to support the upper body by transmitting compressive and shearing forces to the lower body during the performance of everyday activities. To enable the successful transmission of these forces, mechanical stability of the spinal system must be assured. The purpose of this study was to develop a method and to quantify the mechanical stability of the lumbar spine in vivo during various three-dimensional dynamic tasks. A lumbar spine model, one that is sensitive to the various ways that individuals utilize their muscles and ligaments, was used to estimate the lumbar spine stability index three times per second throughout the duration of each trial. Anatomically, this model included a rigid pelvis, ribcage, five vertebrae, 90 muscle fascicles and lumped parameter discs, ligaments and facets. The method consisted of three sub-models: a cross-bridge bond distribution-moment muscle model for estimating muscle force and stiffness from the electromyogram, a rigid link segment body model for estimating external forces and moments acting on the lumbar vertebrae, and an 18 degrees of freedom lumbar spine model for estimating moments produced by 90 muscle fascicles and lumped passive tissues. Individual muscle forces and their associated stiffness estimated from the EMG-assisted optimization algorithm, along with external forces were used for calculating the relative stability index of the lumbar spine for three subjects. It appears that there is an ample stability safety margin during tasks that demand a high muscular effort. However, lighter tasks present a potential hazard of spine buckling, especially if some reduction in passive joint stiffness is present. Several hypotheses on the mechanism of injury associated with low loads and aetiology of chronic back pain are presented in the context of lumbar spine stability. PMID- 11415594 TI - Mechanical behaviour of hamstring muscles in low-back pain patients and control subjects. AB - The purpose of this study was to measure and compare the passive elastic moment, the stiffness and the damping coefficient of the hip joint, as functions of the hip and knee joint angles in men with and without low-back pain. Two conventional tests, the straight-leg-raising test and the trunk forward flexion, were also performed and compared between these subjects. The passive elastic moment was measured using an isokinetic device in the passive mode. This device raised the lower limb from the horizontal position to the straight-leg-raising angle at a slow and constant angular velocity. A custom-made splint connected with the lever arm of the isokinetic device maintained the knee in extension and the ankle in the neutral position. The damping coefficient of the hip joint was measured for 0, 15, 45, 60, 75 and 90% of straight leg raising angle of each subject, using the suspension method based on small oscillation theory. To ensure that muscles were inactive during the passive hip moment tests, muscle activity was monitored with surface EMG. The stiffness was computed as the ratio of the change in passive elastic moment to the change in the hip angle. The passive elastic moment, the stiffness and the normalized trunk flexion were significantly different between the two groups respectively. There was, however, no difference between the two groups in the results of straight-leg-raise and damping coefficient of the hip. The passive elastic moment was a nonlinear function of the hip flexion angle and showed large intersubject differences, especially as the joint limit was approached. The damping coefficient was a polynomial function of the hip flexion angle. The measured variables were analysed using a discriminant function and it was shown that the two groups were clearly discriminable in a meaningful manner. PMID- 11415595 TI - Development and evaluation of a scalable and deformable geometric model of the human torso. AB - A model is developed to provide a geometric representation of the human spine including thoracic and lumbar motion segments, lumbar muscles, ribs, sternum, sacrum, and pelvis. An existing model was modified in order to allow for scaling using standard anthropometric measures, deformation to specific 3-dimensional postures using surface markers, and incorporation of muscle length-tension and motion segment passive bending properties. Experiments were performed to evaluate the accuracy of model postural predictions. Analysis of surface marker displacements demonstrated that the thoracic spine deforms only minimally over a range of flexion, extension, and lateral bending torso postures, suggesting that it can be treated as essentially rigid during low-weight lifting over the range of passive flexibility. Locations of bony landmarks were accurately reproduced (mean errors 2.9-6.8 mm) as were several body dimensions (mean differences 2.6 15.4 mm). It is concluded that linear scaling to subject-specific anthropometry and the use of specific surface markers provides an accurate and direct technique for describing spinal geometry. Predicted passive spinal moments were found to be comparable to those required to support body weight in different extreme postures. It is recommended that data obtained from this type of model be incorporated in future investigations of spinal loading. PMID- 11415596 TI - Properties of musculus gluteus maximus in above-knee amputees. AB - The present study aimed at evaluating possible changes of the gluteus maximus muscle bulk size and in the contracting properties of gluteus maximus in the amputated above-knee limb. Seven male above-knee amputees, with a mean age of 47.4 years volunteered to participate in the present study. Twenty-one healthy subjects with a mean age of 34.0 years served as the control group. Muscle belly displacement was measured by means of a precision electromagnetic sensor dc-dc displacement transducer while performing maximal voluntary isometric contractions. The muscle twitch contraction has been quantified by the latency period (T1), time to peak tension (T2) and the slope of the record. A statistically significant decrease in muscle belly displacement on the amputated side has been observed in comparison with controls and non-amputated side. The latency period on the amputated side was significantly longer and the slope less steep than in the control group. These results indicated atrophied and slower gluteus maximus on the amputated side of above-knee amputees. PMID- 11415597 TI - Three-dimensional kinematics of the rearfoot during the stance phase of walking in normal young adult males. AB - Three-dimensional video analysis was used to quantify three-dimensional motion of the rearfoot in 14 normal young adult males during the stance phase of walking. A stringent calibration procedure enabled standardization of the neutral position of the rearfoot. Surface markers were placed on the calcaneus and lower tibia, and a joint coordinate system was used to determine the relative displacement between the calcaneus and lower leg as representative of rearfoot motion. Confirmation was made of the assumption of a mechanical coupling between the rearfoot displacements of abduction/adduction and eversion/inversion. However, there were several findings which contrasted with commonly held assumptions. For example, the rearfoot was found to evert gradually, rather than rapidly, from heel contact to just prior to heel rise. Also the conventional descriptions of the composite movements of pronation and supination were not applicable to rearfoot movement in this study, and it was therefore concluded that pronation and supination are not valid descriptors of three-dimensional rearfoot motion during the stance phase of walking. PMID- 11415598 TI - The deleterious effect of tetanic contraction on rabbit's triceps surae muscle during cyclic loading. AB - To elucidate the effect of nerve stimulation on the mechanical property of muscle tendon unit, the triceps surae muscle of New Zealand White rabbit was tested with material testing machine. After anaesthesia, the triceps surae muscle-tendon unit of rabbit was tested with cyclic loading during nerve stimulation. The differences between initial and residual force disappeared in the presence of low amplitude and low frequency nerve stimulation. Loading energy applied to the muscle-tendon unit did not change, but the unloading energy increased and energy loss decreased significantly. However, the residual force appeared again at the tetanic stimulation; the loading energy, unloading energy and energy loss all increased significantly. PMID- 11415599 TI - Pedal forces produced during neuromuscular electrical stimulation cycling in paraplegics. AB - This study investigated forces applied to the pedal of a cycle ergometer by paraplegic subjects undergoing neuromuscular electrical-stimulation-induced leg exercise. The patterns of force application were compared with those of able bodied subjects cycling under voluntary muscle control in order to investigate the effectiveness of the muscle stimulation parameters. Results show that paraplegic subjects applied significantly larger peak forces than the able-bodied subjects because of the short duration of neuromuscular stimulation. Able-bodied subjects were able to achieve the same average workload by applying smaller forces over a greater percentage of each crank revolution. It is suggested that the large forces produced by paraplegic subjects contribute to the low efficiency reported by previous studies, and that increasing the range of angles over which muscles are stimulated may provide a means to increase the efficiency of cycling for paraplegic individuals. PMID- 11415600 TI - A computertomographic study on the congruence of the hip joint. AB - This study describes investigations into the appearance of intra-articular spaces of hip joints. In CT scans of cadaver hip joints in the supine position an increased intra-articular space was found dorsally, while in the prone position the same hip joint showed an increased intra-articular space ventrally. It is assumed generally that the gap between the bone ends seen on plane X-rays represents the cartilage, but this study indicates that there may in fact be a space between the cartilage surfaces in the unloaded joint. PMID- 11415601 TI - Epidemiological and aetiological aspects of low back pain in vibration environments - an update. AB - The article reviews the substantial body of epidemiological evidence linking vibration exposure and low back pain. Drivers appear to be at particular risk if exposures exceed those recommended by the ISO exposure limit. Various aetiological factors associated with vehicular vibration, flattening of the lumbar lordosis, increased motion segment flexibility, disc pressure and mechanical softening are discussed. Vibration studies of functional spinal units are also discussed, as are in vivo whole-body vibration experiments. Animal models have shown that vibration leads to compromised nutrition, higher disc pressures, release of neuropeptides, increased creep and histological changes. PMID- 11415602 TI - Non-uniformity of contact stress on polyethylene inserts in total knee arthroplasty. AB - This study examined tibiofemoral contact stresses in 15 commercially available TKR designs, using digitally imaged pressure-sensitive film. The objectives were (1) to determine the correlation between minimization of spatial mean contact stress and minimization of the amount of overloaded (>10 MPa) polyethylene contact area, and (2) to ascertain the difference in contact stresses for machined versus moulded polyethylene inserts. The data showed that there was no statistically significant (design rank-order) correlation of spatial mean contact stress with overloaded polyethylene area. The data also showed that machining cutter preparation of the polyethylene insert caused substantial local contact stress non-uniformities that corresponded to the pattern of grossly visible machining marks. PMID- 11415603 TI - Biarticulating two-dimensional computer model of the human patellofemoral joint. AB - A biarticulating two-dimensional model of the patellofemoral joint has been developed using geometric and force equilibrium constraints to calculate the orientation of the patella about its mediolateral axis. The equations developed from these constraints were solved using an iterative numerical procedure. The model predicts and explains the proximal rolling of the patella on the femur during flexion; it predicts transfer of contact from the trochlea to the femoral condyles at high knee flexion angles. Variation with flexion angle of the patellar mechanism angle and the patellofemoral joint reaction force magnitude agree well with published experimental results. PMID- 11415604 TI - Position and orientation in space of bones during movement: experimental artefacts. AB - This paper deals with the experimental problems related to the reconstruction of the position and orientation of the lower limb bones in space during the execution of locomotion and physical exercises. The inaccuracies associated with the relative movement between markers and underlying bone are analysed. Quantitative information regarding this movement was collected by making experiments on subjects who had suffered fractures and were wearing either femoral or tibial external fixators. These latter devices provided frames that were reliably rigid with the bone involved, and hence the possibility of assessing the relative movement between markers mounted on the skin and this bone. Anatomical frames associated with thigh and shank were reconstructed using technical frames based on different clusters of skin markers and their rotation with respect to the relevant bone evaluated. Marker movement was also assessed in subjects with intact musculoskeletal structures using digital videofluoroscopy. PMID- 11415605 TI - Validation of two 3-D segment models to calculate the net reaction forces and moments at the L(5)/S(1) joint in lifting. AB - The purpose of this study was to examine the validity and sensitivity of two three-dimensional segment models to estimate the net reaction forces and moments at the L(5)/S(1) joint. The two dynamic three-dimensional multisegment models, applied to lifting activities, were a lower body model and an upper body model. Three healthy male subjects participated in this study. The asymmetrical task consisted of lifting a load of 9.6 kg in a normal speed condition and in a fast condition. Results from the two models in terms of joint reaction forces and joint reaction moments at L(5)/S(1) were compared and were then used to validate them. The correlation between the reaction moments from both models were generally above 0.95 and the root mean square (RMS) differences were generally below 10 Nm but could reach 38 Nm. Similar trends were observed in the sensitivity analysis. A proportion of the error was attributable to errors in the segment accelerations because of an increase in the RMS differences between the models with an increase in lifting speed. The use of the lower body model seemed to present some advantages over the upper body model because of the nature of the task analysed which did not require large accelerations from the lower part of the body. PMID- 11415606 TI - Effect of posture on hip joint moment during pregnancy, while performing a standing task. AB - In this paper the combined effect on reactive hip joint moment due to changes in (1) segment mass, (2) trunk centre of mass, and (3) working posture during pregnancy was estimated, and the relative contributions of these three changes to the change in the moment were assessed. The situation studied concerned standing work at a table. Sixteen women were studied monthly during pregnancy. The working posture was assessed by two-dimensional photographic posture recording and description. Body dimensions were measured to assess segment characteristics. A two-dimensional static model was used to estimate the reactive hip joint moment at 10, 20, 30 and 40 weeks of pregnancy. Between 10 and 40 weeks of pregnancy the moment increase was 52 Nm, i.e. the load at 40 weeks of pregnancy is 2.8 times the load at 10 weeks of pregnancy. On average half of this increase was due to postural changes. Changes in segment mass, trunk centre of mass, and the interaction between the three changes accounted for rougly 10, 20 and 15% of the load increase respectively. The increase in reactive hip joint moment may be minimized by preventing adverse postural changes, and optimizing the posture to reduce the contribution of changes in body weight (distribution). PMID- 11415607 TI - Influences of configuration changes of the patella on the knee extensor mechanism. AB - Biomechanical analysis of the configuration changes of the patella was studied. Normal patellar tracking in the sagittal plane was obtained by recording and digitizing knee extension of six healthy adults with fluoroscopy. Based on the digitized data, the changes of patellar thickness were simulated by translating its axis and its forward and backward tilting by rotation in the sagittal plane. We assumed that the extensor moment remained constant before and after the patellar configuration was changed. Therefore, using a balance beam, model equilibrium was reached, after each simulated change of patellar configuration, of the forces involved in the extensor mechanism: the quadriceps muscle force, tension of patella tendon and patellofemoral joint reaction force. The results revealed that when the patellar thickness decreased, quadriceps force increased but patellofemoral joint reaction force decreased and the reverse was seen when the patella thickened. Backward tilting of the patella decreased quadriceps force and patellofemoral joint reaction force and increased the patella tendon/quadriceps force ratio, which produced a better mechanical advantage of force transmission of patella. The reverse was seen in forward tilting. These results suggest that the patella could be tilted backward for the older patient undergoing total knee replacement. For younger patients with relatively strong quadriceps musculature, we suggest that the patellar thickness could be decreased to reduce the patellofemoral joint reaction force so as to reduce the chance of failure of the patella prosthesis. PMID- 11415608 TI - Use of robotics technology to measure friction in animal joints. AB - A new system to measure friction in animal joints has been designed. The system utilizes a robotic arm. Force control of the robotic arm was used to survey the geometry of a sliding surface before the friction of the joint is measured. This enables the precise measurement of friction in joints of complex shape. To test the reliability of the new system we took friction measurements from 10 rabbit stifles. The experimental conditions were: (i) intact joints, (ii) joints after the surfaces were washed. The mean values of the frictional coefficients were 0.008 in the intact joints and 0.015 in the washed joints. These mean values were similar to some previous results obtained from canine hip joints under the same experimental conditions. Furthermore, the standard deviations in this study were smaller than in the previous study. These facts suggest that the new system is reliable. RELEVANCE:--The pathology of osteoarthritis is related to the lubrication mechanisms of joints. In any study of the lubrication mechanisms, an accurate friction measurement is important. In this study we used robotics technology to measure friction in joints. PMID- 11415609 TI - Relationship between alignment and kinematic and kinetic measures of the knee of osteoarthritic elderly subjects in level walking. AB - Eleven osteoarthritic elderly subjects were tested to analyse the relationships between static radiographic alignment and dynamic kinematic and kinetic measures at the knee in gait. A standard precision radiograph procedure (QPR) was used to measure the static lower limb alignment in two planes and a 3D optoelectronic system was used to measure the biomechanical performance of the knee in level walking. Relationships were calculated between the static radiographic angular measures and the dynamic knee angular measures, dynamic knee joint forces, and dynamic knee moments of force during gait. The results indicated that there was not a close relationship between lower limb alignment and the force estimates in gait. However, a closer relationship was observed between QPR parameters, dynamic knee joint angles, and joint moments in gait, implying that complex interrelationships may exist between static alignment, dynamic alignment, and dynamic kinetic measures. These findings suggest that the static alignment measures associated with both the kinematic and kinetic measures should be combined to better explain the biomechanical changes at the knee joint. RELEVANCE:--Values for radiograph alignment and dynamic variables obtained during walking for a group of elderly subjects with moderate osteoarthritis are provided. Joint forces obtained during walking were not related to static radiographic alignment. The findings indicate that both the coronal and sagittal plane measures were associated with the magnitude of joint moments, joint angles, and timing of the events. PMID- 11415610 TI - An isokinetic study of combined activity of the hip and knee extensors. AB - The purpose of this study was twofold: to derive isokinetic concentric and eccentric normal strength values obtained during combined activity of the knee and hip extensors and to compare these values with those based on testing of knee extensors only. Fifteen men took part in the study. Combined extension was tested in the supine position with the knee and hip at an initial position of 95 degrees. Knee extension was tested in the seated position with the knee at the same initial position. The range of motion and angular velocities were equal in both configurations: 15 degrees and 8 degrees s(-1) and 15 degrees s(-1) respectively. The average peak force in the CKH mode was close to 10 Nm/kg bw and over 15 Nm/kg bw in the concentric and eccentric contractions respectively, whereas for the knee extensor mode the respective figures were approximately 8 Nm/kg bw and 10 Nm/kg bw. With respect to the same performance parameters, there were low and insignificant correlations between the two modes. RELEVANCE:- Isokinetic testing of knee extensors performance in what is commonly known as single-joint configuration is somewhat limited in its functional scope and, if applied wrongly, is potentially harmful in various dysfunctional states of the knee. On the other hand, multi-joint testing configuration, involving muscles of the hip as well as the knee, is possibly more functional and safe. To assess the muscle performance status of knee patients using the latter mode, data regarding what should be expected from normal individuals is essential. This paper supplements existing material concerning knee and hip extensors and looks into the relationship between the two dynamic contraction modes as well as the association between combined activity and knee extensors only. This association which was found to be very limited indicates that the two configurations reflect different muscle action paradigm and therefore may not be used interchangeably. PMID- 11415611 TI - Trunk muscle parameters determined from MRI for lean to obese males. AB - Transverse magnetic resonance images at the L(4)-L(5) level of 26 males ranging in body mass index from 19.7 to 39.5 were digitized using a computer video monitor with mouse-mediated digitization software. The digitized musculature included the right and left psoas, rectus, abdominis, quadratus lumborum, oblique, and paraspinal muscle groups. From the digitized profiles, cross sectional areas, and anteroposterior and mediolateral moments from the vertebral centroid were subsequently calculated. Multiple regression formulae were derived to predict the muscle parameters from the most strongly related anthropometrics. Cross-sectional area and moment arms for lean and obese groups were also compared. Significant regression formulae were derived for the cross-sectional area of the rectus abdominis and obliques, for mediolateral moment arms of the paraspinals, psoas, and obliques, and for anteroposterior moments of all four muscles. Comparison of muscle parameters for lean and obese subjects revealed minimal differences, though normalization of muscle cross-sectional areas by trunk area produced values which were statistically smaller for all muscles of the obese relative to lean subjects. RELEVANCE:--Identifying variations in trunk muscular geometry across specific population groups will assist in further refining the existing morphometric data base used to estimate the forces experienced about the lower back. By identifying these variations, it may be possible to observe factors which may predispose a specific population to low back pain. PMID- 11415612 TI - Trunk and hip muscle recruitment in response to external anterior lumbosacral shear and moment loads. AB - The response of select lumbar torso and pelvic muscles to loads applied at the lumbar spine and across the pelvis was investigated to help identify the sources of antagonistic lower torso muscle recruitment. Five subjects in the neutral upright posture were loaded with L(4-5) moment and anterior shear forces under constrained and free pelvic rotation conditions. The activity states of the erector spinae, latissimus dorsi, external oblique, rectus abdominis, gluteus maximus, and rectus femoris muscles were monitored using surface electromyography. Regression analysis was used to test if (1) lumbar shear loads influence the activity of the torso muscles, and (2) moments about the pelvis significantly influence the torso muscle activities. The pelvic muscles (gluteus maximus and rectus femoris) were monitored for their role in stabilizing pelvic rotation. The statistical analysis did not support the hypothesis that torso muscles are recruited in response to shear loading. Also, the data did not provide conclusive evidence that moments about the pelvis relative to the lumbar spine significantly influence torso muscle recruitment. For the subjects that exhibited elevated antagonistic muscle activity, no explanation of this recruitment strategy could be statistically related to lumbar shear or pelvic moments. RELEVANCE:--Omission of pelvic moments in biomechanical modelling of the lumbar torso does not seem to be introducing significant error in the prediction of torso muscle activity. The results of this study indicate that the source of lumbar torso muscle antagonistic activity under sagittal plane flexion loading seems to be a result of yet unidentified factors. PMID- 11415613 TI - Sit to stand from progressively lower seat heights -- alterations in angular velocity. AB - This study investigates the influence of chair height on the dynamics of sit-to stand for two age groups. Eleven young (25-36 years) and 10 older (61-79 years) adults participated. Subjects rose from chairs set at four heights relative to knee height. Motion was quantified using a bilateral active-marker-based motion analysis system. Subjects appeared to increase trunk flexion angular velocity to overcome mechanical difficulties of decreasing chair heights. This variable showed a main effect for chair height (P = 0.0001). Time at which knee, hip, and trunk extension angular velocity were attained each demonstrated a chair by age interaction effect (P<0.05). Synchrony of body segment maximum extension angular velocities was altered for the older subjects at the lowest chair heights, suggesting that older individuals begin to change their performance as the task becomes more demanding. RELEVANCE:--Sitting to standing is one of the essential physical tasks used frequently throughout the day. Clinicians are frequently called upon to improve chair rise performance for those with functional limitations. Efforts are likely to be most successful if clinicians understand how healthy individuals accommodate to changing conditions (such as changing chair height) and use that information to interpret the performance of those with impairments and functional limitations. PMID- 11415614 TI - Generalizability of in-shoe peak pressure measures using the F-scan system. AB - In-shoe pressure analysis can be useful in the management of a variety of foot and ankle problems, but guidelines are needed to determine the practical limitations of the measures. The primary purpose of this study was to determine the reliability of peak plantar pressures taken with the F-Scan system over multiple steps, sensors, and days, and using a force platform for additional calibration. Data were collected on 10 healthy subjects as they walked across a force platform for a minimum of three trials on four separate sessions that were 1 week apart. Using a mean of three steps with a single sensor on 1 day, generalizability coefficients were 0.75 for manufacturer calibration and 0.82 with force platform calibration; reliability coefficients for absolute decisions (index of dependability) were 0.60 for manufacturer calibration and 0.76 with force platform calibration. Force measures from the F-Scan and force platform were highly correlated (r = 0.93), but the absolute difference between the measures varied between sensors and over time. RELEVANCE:--Peak pressure measures taken from a mean of three steps with procedures outlined by the F-Scan manufacturer provide pressure values that demonstrate adequate reliability for clinical and research purposes when rank ordering of measures is indicated. If measures are to be used for making absolute decisions (e.g., identifying a threshold for injury), calibration from a stable source such as a force platform is recommended. PMID- 11415615 TI - Technique for the measurement of hindfoot inversion and eversion and its use to study a normal population. AB - Following earlier work identifying the difficulties of defining the subtalar neutral position, the authors have developed new techniques to measure the ranges of inversion and eversion in normal subjects. A test rig has been designed and constructed to apply a known torque about an anterior posterior axis and an electrogoniometer has been used to measure the resulting movements. The neutral position was defined as that position corresponding to the footplate being horizontal and the leg vertical. In a study of 100 normal subjects (50 female/50 male), it has been shown that the average overall range of motion decreased from 53.2 degrees in 20-year-olds to 44.3 degrees in 60-year-olds. It was also demonstrated that females had a greater range of motion than males, but that this difference reduced with age. This fact was largely accounted for by a decrease in the range of eversion in female subjects. RELEVANCE:--Hindfoot motion is essential for human walking, but accurate measurement is difficult because of the structure of the joints involved. This paper describes the development of a technique using electrogoniometry, for carrying out measurement of passive inversion and eversion of the hindfoot in a sample of healthy adults. It further highlights the difficulties in carrying out an accurate evaluation of hindfoot motion on which to base clinical decisions. PMID- 11415616 TI - Quantitative intramuscular myoelectric activity of quadratus lumborum during a wide variety of tasks. AB - Intramuscular fine-wire electrodes monitored the electromyographic activity of quadratus lumborum in four young adults. A wide variety of tasks were performed including flexion tasks, lateral bending, twisting, extension, and lifting tasks. Heavy lifts of barbell weights up to 70 kg activated the quadratus lumborum 74% of their maximum on average while surface recording of erector spinae (L(3)) were only 62% of their maximum activation. The quadratus lumborum was more active (54%) than other muscles during isometric side support postures where the body is held horizontally almost parallel to the floor as the subjects supported themselves on one elbow on the floor together with both feet. Furthermore, it increased activation in response to increasing compression in static upright standing postures. RELEVANCE:--Electromyographic evidence, together with architectural features make the quadratus lumborum a better stabilizer of the spine than psoas. Use of horizontal 'side support' exercise to train this muscle would appear to be a wise choice. PMID- 11415617 TI - An ultrasonic-operated kinematic measurement system for assessment of stance balance in the clinic. AB - This study describes the application of a kinematic measurement system based on ultrasonic technology to enable the assessment of stance balance in the clinic. The system is composed of 1-8 ultrasonic transducer markers which transmit ultrasonic waves, and three ultrasonic receivers. The spatial position of each marker is monitored by calculation of the time interval between transmission and reception of the wave. Sway data from the waist of 44 healthy subjects and 15 hemiparetic patients were collected during quiet stance. The results distinguished between data collected with and without vision, between patients and controls, and between the two sides of the body. The findings were found to be valid and reliable in repeated measurements. RELEVANCE:--Assessment of stance balance in the clinic is a requisite for quantifying disability and measuring improvement in patients with postural control deficits. Due to high costs and complexity of operation, the use of prevailing equipment for these purposes is mainly confined to research facilities. This paper describes a unique kinematic measurement system whose low price and ease of operation make it feasible for clinical use. PMID- 11415618 TI - Postural stability measures: what to measure and for how long. AB - This paper assesses the repeatability of force plate postural stability measurements. The common parameters were investigated for five test durations, two vision conditions, and two stance conditions. All parameters were found to be reliable measures. Optimum test retest reliability was obtained at 20-s and 30-s trial periods. Centre of pressure measures increased with increased test duration while ground reaction forces and velocity decrease slightly. RELEVANCE:--The results indicate that measures of centre of pressure excursion and ground reaction forces can differentiate between various mechanical and sensory conditions and thus are clinically useful for detecting and quantifying both mechanical and sensorimotor postural deficiencies. It is also demonstrated that the test duration is important in the reliability and validity of stability measures. PMID- 11415619 TI - The clinical biomechanics award paper 1995 Lower extremity joint loading during impact in running. AB - OBJECTIVE: The main purpose of this study was to estimate lower extremity joint impact loading in running and the influences of muscles on this loading. DESIGN: A 2D simulation model that included skeletal motion, muscles, and soft-tissue movement was developed in this study. BACKGROUND: Our understanding of joint impact loading has been mainly based on measurable external loading variables. Furthermore, changes in muscular forces have often been neglected, assuming that muscular activation cannot react during the period of impact, which ignores passive muscle properties. METHODS: Kinematics and ground reaction forces were collected for each of five subjects performing heel-toe running. Kinematic and inverse dynamics analyses provided the initial conditions for the simulation models. The motion of each subject was simulated for 50 ms following heelstrike. RESULTS: Motion of body segments following heelstrike reduced the rate of joint impact loading, sometimes substantially, in comparison to the ground reaction force. In addition, substantial changes in muscular forces were sometimes observed that further reduced the rates of joint loading. CONCLUSIONS: The rates of joint impact loading are reduced in comparison to the ground reaction force during normal heel-toe running. Changes in muscular forces may have a relevant effect on joint impact loading and should not be neglected in future studies. RELEVANCE: Repetitive external impact loading has resulted in degeneration of articular cartilage in animal models. However, runners, as a group, do not have a high incidence of osteoarthritis. The present study suggests that there are mechanisms available to the runner that can substantially reduce the rate of joint impact loading relative to the rate of external loading. These mechanisms may be sufficient to prevent overloading and degeneration of the joints. PMID- 11415620 TI - Time-dependent changes in the lumbar spine's resistance to bending. AB - OBJECTIVE: To show how time-related factors might affect the risk of back injury. DESIGN: Mechanical testing of cadaveric lumbar motion segments. BACKGROUND: High bending stresses acting on the lumbar spine are associated with injuries to the intervertebral discs and ligaments. Since these soft tissues are viscoelastic, the bending stress ('bending moment') must depend on the speed of movement and the duration of loading, but this has not previously been quantified. METHODS: Forty-five cadaveric lumbar segments, consisting of two vertebrae and the intervening disc and ligaments, were loaded in combined bending and compression in order to simulate movements and postures in living people. The relationship between flexion angle and bending moment was determined at different loading rates, and after sustained loading in bending and in compression. RESULTS: Rapid flexion movements increased the peak bending moment by 10-15% compared to slow movements. On average, repeated flexion over a period of 5 min reduced the peak bending moment by 17%, and 5 min of sustained flexion reduced it by 42%. Two hours of compressive creep loading reduced the height of the intervertebral discs by 1.1 mm, increased the range of flexion by 12%, and reduced peak bending moment by 41%. CONCLUSIONS: The scale of these changes suggests that, in life, the risk of bending injury to the lumbar discs and ligaments will depend not only on the loads applied to the spine, but also on loading rate and loading history. RELEVANCE: The results show how time-dependent factors can increase the risk of bending injury to the osteoligamentous lumbar spine. PMID- 11415621 TI - Three-dimensional analysis of active cervical motion: the effect of age and gender. AB - OBJECTIVE: To describe the effects of age and gender on three-dimensional (3D) active cervical spine motion. DESIGN: This was a descriptive study. BACKGROUND: This study expanded on previous investigations of age and gender effects on single plane motion of the cervical spine. METHODS: Sixty female and 60 male asymptomatic, normal volunteers, aged between 20 and 59 years, were examined in a standardized seated position. The 3 SPACE Isotrak system was used to measure simultaneous 3D motion of the cervical spine. RESULTS: The mean range of all the primary movements decreased significantly with age. For flexion/extension the greatest decrease occurred between the 20- and 30-year-olds, whereas for both lateral flexion and rotation, significant differences were demonstrated in subjects aged two decades apart. The coupling of motion associated with rotation was significantly related to age. CONCLUSION: Age had a significant effect on all of the primary movements. Age had less effect on the range of the coupled movements, in that the only movements to be affected were lateral flexion and extension occurring during cervical rotation. Gender had no marked effect on the primary or the coupled movements. RELEVANCE: This study provides normative data for the effects of age and gender on three-dimensional analysis of active cervical spine motion, which can be used for comparison with specific patient populations. The high level of intra-subject test-retest reliability renders the3 SPACE system of value for clinical measurement of movement pre- and post treatment intervention for cervical spine disorders. PMID- 11415622 TI - Biomechanical and histological changes in the patellar tendon after in situ freezing An experimental study in rabbits. AB - OBJECTIVE: The effects of freezing on the remodelling process of the patellar tendon were examined. DESIGN: An experimental study in rabbits. BACKGROUND: Patellar tendon weakens when grafted as a subsstitute for the anterior cruciate ligament. Fibroblast necrosis is considered to be one of the many factors contributing to this change. Therefore, the effect of freeze-induced necrosis on the patellar tendon has been studied. METHODS: Using a technique for freezing the patellar tendon in situ with liquid nitrogen to kill fibroblasts, we studied the biomechanical and histological changes in the patellar tendon up to 24 weeks after freezing. RESULTS: The cross-sectional area started to increase by week 3, reaching a plateau by week 12. The elastic modulus and tensile strength began decreasing by week 3. Although the maximum load decreased at weeks 12 and 24, the stiffness did not change. Histologically, cells were absent until week 2. Athough cells were apparently normal at week 24, there were none of the dense collagen bundles that are normally seen. CONCLUSIONS: The once-frozen patellar tendon weakens as tissue remodelling occurs. RELEVANCE: The study was designed to ascertain whether the remodelling process of the once-frozen patellar tendon coincides with its mechanical properties in different phases. The weakening of the patellar tendon occurred as new cells proliferated into the tendon and remodelled the tissue. PMID- 11415623 TI - An experimental study of damage accumulation in cemented hip prostheses. AB - OBJECTIVE: To develop a methodology to characterize the pattern of crack initiation and damage accumulation in intramedullary fixated cemented prostheses. DESIGN: An experimental physical model of intramedullary fixation was developed which both represents the implant structure and permits monitoring of fatigue crack growth. BACKGROUND: Many joint replacement prostheses are fixed into the medullary cavity of bones using a poly(methylmethacrylate) 'bone cement', which forms a mantle around the prosthesis and locks it to the bone. The endurance of the replacement is, to a great extent, determined by the mechanical durability of the cement and the implant interfaces under cyclic stresses generated by dynamic loading. The cement mantle is subjected to complex multiaxial stresses which vary in particular distribution depending on the prosthesis design. METHODS: Damage accumulation is reported in terms of the number of cracks, the location of cracks, and the rate of crack growth. RESULTS: The results clearly show the nature of damage accumulation in the cement mantle, and that many of the cracks which propagate within the cement mantle are related to cement porosity. CONCLUSION: This study gives experimental evidence to support the hypothesis of a damage accumulation failure scenario in cemented hip reconstructions. RELEVANCE: Cementing is the most popular technique for the fixation of joint replacement prosthesis. However, the sequence of events leading to the failure of cemented fixation is not fully understood. In this paper it is shown that damage accumulation can be directly monitored in an experimental model of cemented intramedullary fixation. PMID- 11415624 TI - Scapho-lunate dissociation and arthrodesis. An experimental study with lesions of the interosseous ligament and fusions with K-wires. AB - The carpal mechanism is studied during a sectioning of the ligament between the lunate and the scaphoid. A tear in this ligament is frequently observed in the clinic. After experimental lesions of this ligament, some authors say there is little effect while others report significant changes. We have conducted repetitions of these sectioning experiments using roentgen stereophotogrammetry to assess the kinematics of the carpal bones. We have found very little effect. We have also fused the scaphoid to the lunate to find out what happens to the carpal mechanism when the ligament between these bones is made stiffer. Fusion is carried out with K-wires and the effect upon the carpal mechanism is studied again. Our K-wiring of the scaphoid and lunate shows that the carpus starts to behave unpredictablely. The positions of the wires influence the outcome significantly. RELEVANCE--:The ligament between the scaphoid and the lunate has been assigned a main function in the carpal mechanism. It is still in debate, however, whether a lesion of this ligament by itself leads to the typical scapho lunate dissociation pattern of the carpus. We have shown that a disruption of this ligament hardly influences the carpal mechanism. In time though, other ligaments may become overstretched so that finally the typical pattern may show up. K-wiring of the lunate and scaphoid to restore the connection between these bones results in a primarily unstable joint. PMID- 11415625 TI - Rotational laxity and stiffness of the radiocarpal joint. AB - OBJECTIVE: To investigate the constraint and potential mechanism of torque transmission across the wrist joint. DESIGN: In vitro experiment using human cadaveric specimens. BACKGROUND: Transmission of torque from the forearm to the hand requires rotational stability at the wrist. Better appriciation of the constraints would have applicability to several clinical problems where the stability is compromised. METHODS: Thirteen fresh-frozen cadaveric specimens were used in this experiment to investigate the rotational laxity and stiffness of the radiocarpal joint in unloaded and axially loaded (100 N) conditions, and three forearm orientations in a neutral, pronation (60 degrees ), or supination (60 degrees ) position. RESULTS: In pronation or supination, there was no difference between loaded and unloaded conditions in primary or total laxity at a maximum torque of 2.3 Nm. Unloaded specimens showed a mean total rotational laxity of 42.1 degrees. Supination or pronation of the forearm caused a decrease in laxity with respect to neutral forearm rotation (35 degrees and 41.6 degrees versus 49.6 degrees respectively). The primary rotational laxity accounted for half of the total laxity. With axial compression, total rotational laxity did not change, but primary laxity dropped to 50% of its unloaded value. The primary stiffness was very low -- approximately 11% of the secondary stiffness. CONCLUSION: The ligamentous structures and the joint articulation restricted excessive axial rotation of the wrist. However, a laxity of approximately 20 degrees was identified for normal wrists. RELEVANCE: This study demonstrated that the primary axial rotational laxity of the radiocarpal joint was approximately 20 degrees. In the mathematical model and implant design, muscular balance of the joint within such laxity should be considered. PMID- 11415626 TI - Isokinetic and isometric strength of the thigh muscles in below-knee amputees. AB - OBJECTIVE: The strength of the quadriceps and hamstring muscles of the amputated limb in below-knee amputees was evaluated. DESIGN: A descriptive study based on clinical measurements of muscle strength. BACKGROUND: The residual limb of below knee amputees is less active in daily functions. As a result, atrophy of the thigh muscles in the affected limb is a common finding. Quality of standing and gait performances depend on the strength of these muscles, which activate the knee of the amputated limb. METHODS: Isokinetic concentric and eccentric, as well as isometric, strength of thigh muscles was measured by means of a dynamometer. Parameters of peak torque and maximal average torque of the quadriceps and hamstring muscles were considered. RESULTS: Significant decreases (P < 0.01) in strength of the quadriceps and hamstrings were observed in the amputated limb. Thigh muscle strength in amputees over 7 years was not significantly weaker when compared with amputees where amputation had been performed more recently. CONCLUSIONS: In below-knee amputees, the thigh muscle strength of the amputated limb decreases significantly during the first years after amputation. RELEVANCE: In so far as the ultimate goal in rehabilitation of amputees is to return the patient to an acceptable level of function, it is recommended that the amputee should be trained and encouraged in self-strengthening exercises for the thigh muscles of the amputated limb. PMID- 11415627 TI - Body height changes with hyperextension. AB - OBJECTIVE: To automatize the lumbar physical examination with an acceptable rate of error. DESIGN: An external skin marker method for automatizing the physical examination was developed and its ability to discriminate between normal and abnormal subjects tested in a blind clinical trial. BACKGROUND: The low reproducibility of clinical findings, even among experienced doctors, has been well documented. This is of particular concern and may explain why there is such a wide variation in surgical rates across the USA (tenfold for disc herniation). Inconsistencies among physicians in the evaluation of benign low back conditions make standardization desirable. METHODS: A computerized physical examination was used to evaluate patients with low back pain and compare their results with a normative database obtained from a selection of healthy subjects. A high resolution motion analysis system tracked the movement of skin markers placed on the midline and pelvis. Surface EMG electrodes placed above L(5) collected data from multifidus. From the kinematics of skin markers during flexion--extension with lifts up to 32 kg, and lateral bending with lifts up to 10 kg, the following parameters were estimated: lumbosacral angle and elongation, contribution of each lumbar segment to the lordosis reduction, relative pelvic/spine motion, and trunk velocity. First the average normal value for each estimated parameter was determined using 40 normal subjects. For each subject the difference between his parameter and the normal was processed by an expert system generating a normality index varying from zero (perfect abnormal) to one (perfect normal). To develop the expert system's rules, a preliminary group of 20 very abnormal subjects was used, such that the normality index separated them from the normals. For validation, a set of 29 back-sprain patients and another set of 42 discogram positive patients were selected. Each subject was tested and his computerized normality index calculated without any clinician's input, then compared with the clinician's evaluation, which was taken to be the gold standard. The receiver operating characteristic technique was used to quantify the discrepancies. RESULTS: The expert system could detect clinically abnormal subjects with accuracy (sensitivity 83-91% and specificity > 90%) whele providing quantitative information on workers' functional capacities. CONCLUSIONS: Once a reference normative database is agreed upon, each patient can be compared with that reference according to the same rules, with the resulting machine classification being independent of the clinician. This eliminates the inter- and intra clinician variability in patient follow-up. Because of the severity of the selection criteria, this study is based upon a relatively restricted number of subjects, as well as a limited normative database of 40 subjects. RELEVANCE: It is possible to automate the lumbar physical examination with an acceptable error rate. This technique permits the objective consistent assessment of lumbar function, and thus allows the comparison of different treatment regimes for lumbar dysfunction. PMID- 11415628 TI - Biomechanics of longitudinal arch support mechanisms in foot orthoses and their effect on plantar aponeurosis strain. AB - OBJECTIVE: The purpose of this investigation was to quantify the longitudinal arch support properties of several types of foot orthosis. DESIGN: An in vitro method that simulated 'static stance' was used to determine arch support capabilities, with plantar aponeurosis strain implemented as the performance measure. BACKGROUND: A longitudinal arch support mechanism of an orthosis resists depression of the foot's arches by transferring a portion of the load to the medial structures of the foot. Since the plantar aponeurosis is in tension when the foot is loaded, a quantifiable decrease in strain should occur with an adequate orthotic arch control mechanism. METHODS: A differential variable reluctance transducer was surgically implanted in the plantar aponeurosis of cadaveric donor limb feet (n = 7). Each specimen was mounted in an electromechanical test machine which applied a load of up to 900 N axially to the tibia. The test schedule was divided into seven test conditions: specimen barefoot; specimen with shoe and specimen with shoe and five different orthoses. RESULTS: The University of California Biomechanics Laboratory Shoe Insert and two other foot orthoses significantly decreased the strain in the plantar aponeurosis compared to the barefoot control and were considered effective arch supports (P < 0.05). The functional foot orthosis, stock orthosis, and test shoe did not effectively reduce plantar aponeurosis strain. Significant variations of time required to achieve the specified load levels were recorded among the test conditions, indicating the relative cushioning properties of the shoe/orthosis systems. CONCLUSIONS: The patterns of plantar aponeurosis strain observed in cadaveric tests suggest that certain types of orthoses are more effective than others in the support of the foot's longitudinal arches. It is suggested that to support the longitudinal arches of the foot effectively the medial surface contours of the orthosis must stabilize the apical bony structure of the foot's arch. RELEVANCE: Reducing tension in the plantar aponeurosis is an important treatment objective for orthotic management of plantar fasciitis. Therefore it is of great clinical interest to know whether the longitudinal arch support mechanism of specific foot orthoses have benefits with respect to the loading of the plantar aponeurosis. PMID- 11415629 TI - Relationship between vertical ground reaction force and speed during walking, slow jogging, and running. AB - OBJECTIVE: To obtain descriptive information between vertical ground reaction force (GRF)-time histories and gait speed, running style, and gender. DESIGN: GRF time history measurements were obtained from male and female subjects during walking, slow jogging, jogging and running on an indoor platform. BACKGROUND: Previous studies have established GRF descriptor variables for male subjects running at speeds from 3 to 6 m s(-1), but very little descriptive data exists for slower or faster running, nor have previous studies reported GRF descriptors separately for female subjects. METHODS: GRF-time histories were recorded for 13 male and 10 female recreational athletes during walking and slow jogging at speeds between 1.5 and 3.0 m s(-1), and running at speeds between 3.5 and 6.0 m s(-1). Vertical GRF-time data for trials with speeds within 0.2 m s(-1) of the prescribed speed were analysed to determine thrust maximum GRF (F(z)) and loading rate (G(z)). RESULTS: In both male and female subjects, F(z) increased linearly during walking and running from 1.2 BW to approximately 2.5 BW at 6.0 m s(-1), remaining constant during forward lean sprinting at higher speeds. F(z) was linearly correlated to G(z), the latter ranging from 8 to 30 BW s(-1) over this speed range. Slow jogging was associated with a > 50% higher F(z) and G(z) in comparison to walking or fast running. CONCLUSIONS: Similar GRF descriptor data and velocity relationships were obtained for male and female subjects. Impact forces were greatest when the subjects adopted a higher, less fixed centre of gravity during slow jogging. RELEVANCE: These results suggest that vertical GRF norms can be established for male and female subjects alike, and that slow or fast running with a lower, fixed centre of gravity decreases impact forces. PMID- 11415630 TI - Computer-aided three dimensional assessment of knee-joint cartilage with magnetic resonance imaging. AB - OBJECTIVE: An MRI-based technique for non-invasive assessment of the quantitative distribution of articular cartilage in the knee-joint was to be developed, and its accuracy and reproducibility tested. DESIGN: Three cadaveric specimens and one patient were studied and MRI measurements compared with anatomical sections or arthroscopy. BACKGROUND: Data on articular cartilage thickness is needed for the design of computer models, determination of cartilage material properties from arthroscopy and staging of osteoarthrosis. METHODS: The knees were imaged using strongly T2-weighted spin-echo and FISP-3D sequences. After digital subtraction and automatic segmentation, three-dimensional reconstruction of the cartilages was performed. Surface areas, volumes and the mean cartilage thickness were calculated, and the regional distribution displayed after trigonometric correction. RESULTS: The difference between MRI volumes and those obtained from the sections ranged from 4 to 21% with a reproducibility of +/-4 to +/-12% after repositioning. The thickness maps obtained with MRI were very similar to those from the sections. In the patient, a full-thickness defect demonstrated with MRI was verified by arthroscopy. CONCLUSIONS: Using the technique presented, the quantitative distribution of knee-joint cartilage may be analysed non-invasively, accurately, and in a very time-effective manner, in cadavers and in living subjects. RELEVANCE: To date there exists no accepted method for the accurate, fast and non-invasive assessment of articular cartilage thickness. Such a technique is, however, very helpful for generating computer models of diarthrodial joints, determination of cartilage material properties during arthroscopy, staging of joint disease, and objective control of chondroprotective treatment. PMID- 11415631 TI - Intra-abdominal pressure during trunk extension motions. AB - OBJECTIVE: This study was designed to help interpret the biomechanical role of intra-abdominal pressure during lifting type motions of the trunk. DESIGN: An in vivo study was performed in which intra-abdominal pressure was observed as subject trunks were subjected to different dynamic trunk loading conditions common during industrial lifting. BACKGROUND: There is a little consensus as to the biomechanical role of intra-abdominal pressure during lifting. Previous studies have suggested that: it may assist in load relief when lifting, may be involved in trunk stability, and/or may be used as a measure fo spine loading. Thus, in general, our understanding of intra-abdominal pressure is rather poor. METHODS: In this study intra-abdominal pressure was monitored using a radio pill in 114 subjects over a series of four experiments. Subject's trunks were subjected to different dynamic trunk symmetric and asymmetric trunk loading conditions that are common during industrial lifting tasks. RESULTS: The results indicated that (1) intra-abdominal pressure increased to significant levels (above 10 mmHg) only when more than 54 Nm of trunk torque were supported; (2) intra-abdominal pressure increases monotonically (up to 150 mmHg) as a function of trunk velocity; and (3) under concentric conditions intra-abdominal pressure increases as a function of greater asymmetry, whereas, under eccentric conditions the response changes to a much lesser extent as asymmetry changes. CONCLUSIONS: These findings suggest that intra-abdominal pressure appears to be more a by product of trunk muscle coactivation. Any mechanical advantage gained from intra abdominal pressure might be in the form of a preparatory action resulting from muscle coactivation that stiffens the trunk just prior to a rapid trunk extension exertion. This function may reinforce previous hypotheses regarding the stability role of intra-abdominal pressure. RELEVANCE: Intra-abdominal pressure has been observed during lifting for several decades, yet the biomechanical role of intra abdominal pressure is poorly understood. This study has attempted to describe how intra-abdominal pressure behaves during lifting motions as the components of lifting are changed. The findings place in doubt biomechanical significance of intra-abdominal pressure. Thus, based upon this study, clinicians need not worry about interpreting intra-abdominal pressure, since it appears to be a by-product of muscle contraction and cocontraction. PMID- 11415632 TI - The use of artificial neural networks to identify patients with chronic low-back pain conditions from patterns of sit-to-stand manoeuvres. AB - OBJECTIVE: To investigate whether artificial neural networks (ANNs) can categorize healthy subjects, chronic low-back-pain (LBP) patients, and subjects pretending to have low-back pain problems, based upon patterns of stand-sit-stand manoeuvres. DESIGN: A non-invasive laboratory study of human subjects. BACKGROUND: Normal strategies for sit-stand manoeuvres are modified in cases of chronic LBP. Subtle changes and many parameters are unsuitable for conventional statistics. METHODS: Fourteen healthy subjects, 10 chronic LBP patients, and 12 subjects pretending to have LBP participated. Forces and centres of pressure at the feet and knees, plus hip and lumbar movements provided inputs into a three layer feed-forward ANN with sigmoidal transfer functions. The ANN was trained with data from 35 of the 36 subjects, and its ability to classify the left-out subject was tested. This was repeated with each subject omitted from training in turn. RESULTS: The ANN correctly classified 31 of 36 subjects. The subjects were also classified by nine physiotherapists from videos of the manoeuvres. Their success rate was significantly lower that that of the ANN, which is not surprising for an unusual procedure without training. CONCLUSIONS: ANNs should be considered as additional tools in assessment and possible diagnosis of pathological movements. RELEVANCE: The capacity of ANNs to discriminate patterns of human movement offers a technique that may prove to be a useful tool for assessment and diagnosis. PMID- 11415633 TI - Spinal compression at peak isometric and isokinetic exertions in simulated lifting in symmetric and asymmetric planes. AB - OBJECTIVE: To determine if the spinal compression can be used as a criterion for the safety of the back. DESIGN: Various lifting strength activities were analysed using a 3-D biomechanical model to determine spinal compression forces. BACKGROUND: Despite many standards being set and the use of varied strategies, low-back injuries continue to be common in our society. Since the injuries occur to the tissues in the body, it may be more appropriate to investigate the effects of external loads on the tissues. Therefore it was decided to determine the spinal compression in standardized lifting tasks. METHODS: Twenty young adults (12 males and 8 females) performed maximal stoop and squat lifts in sagittal, 30 degrees lateral, and 60 degrees lateral planes at half, three-quarters, and full horizontal reach distances. The stoop lifts were performed in isokinetic and isometric modes; the isometric mode was performed with the hip at 60 degrees and 90 degrees. The squat lifts were also performed in isokinetic and isometric modes; the isometric mode consisted of postures with knee at 90 degrees and 135 degrees of flexion. In addition, the subjects also performed isometric lifts in stoop and squat postures at a self-selected optimum posture. During these activities the strength was measured using a static dynamic strength tester employing a load cell and force monitor. Three-dimensional postural recording was made using a 3-D Peak Performance Technologies imaging system. Using the postural and force data as input to a 3-D biomechanical model, the lumbosacral spinal compression was calculated. The values of strength and spinal compression were analysed and compared. RESULTS: The strength was significantly affected by the gender, the type of lift, plane of lift and reach of lift (P < 0.01); whereas the spinal compression was not affected by the reach at all. Two-way and three-way interactions were significant for strength (P < 0.02) but not for spinal compression. CONCLUSION: It was concluded that the spinal compression was the ultimate cost function in the safety of the back and as such preventative strategies should be based on this. RELEVANCE: The use of an appropriate cost function in a manner in which it can maximize the safety of the back will be of considerable value. Since injuries are biomechanical perturbation of the tissues, it is argued that the cost function be used in a manner where tissue load be considered as the primary factor. PMID- 11415634 TI - Biomechanical aspects of plating osteosynthesis of transverse clavicular fracture with and without inferior cortical defect. AB - OBJECTIVE: To find out whether superior or anterior plating osteosynthesis of clavicular fractures with and without inferior cortical defect provides more stability. DESIGN: The bearing of maximal bending moment of the osteosyntheses was determined by applying a load from a universal testing machine. BACKGROUND: A plate is recommended for internal fixation of the clavicular fracture. Stability of the osteosynthesis depends on placing of the plate at anterior or superior aspect of the clavicle. Whereas, the pattern of the fracture also influences stability. METHODS: Pairs of fresh cadaveric clavicles were used. The fractures were created at middle clavicle. Both superior and anterior plating fixation of both patterns of the fractures were performed. A compression load was applied at lateral end of the clavicle and maximal bending moment was calculated. RESULTS: For fractures without inferior cortical defect, the superior plating could withstand 12.05 (SD, 1.74) Nm and 8.69 (SD, 1.56) Nm for the anterior plating. For the fractures with inferior cortical defect, the superior plating could withstand 7.87 (SD, 2.58) Nm, but 10.26 (SD, 2.19) Nm for the anterior plating. CONCLUSIONS: The superior plating of the fracture without inferior cortical defect provides more stability against the bending moment than the anterior plating (P = 0.008). Whereas, the anterior plating of the fracture with inferior cortical defect provides more stability (P = 0.025). RELEVANCE: When plating a clavicular fracture without inferior cortical defect is performed, the plate should be placed at superior aspect of the clavicle for achieving more stability of the osteosynthesis against the bending moment. On the other hand, when the fracture has an inferior cortical defect, the plate should be placed at the anterior aspect of the clavicle. PMID- 11415635 TI - Direct measurement of hoop strains in the intact and torn human medial meniscus. AB - OBJECTIVE: To measure the circumferential or hoop strains generated in the medial meniscus during loading of the knee joint and to examine the effect of longitudinal and radial tears in the meniscus on these strain values. DESIGN: An in vitro investigation measuring the circumferential strains in the medial menisci of cadaveric human knees as they were loaded in a materials testing machine. BACKGROUND: The menisci transmit approximately 50% of the load through the knee, the rest being transmitted by direct contact of the articular cartilage. Damage to the menisci will alter the pattern of load transmission as will meniscectomy. This study examined the changes in the mechanics of the meniscus in situ as a result of simulated tears to assess the effect of its load carrying capacity and the implications of surgery to remove part or all of a damaged meniscus. METHODS: Nineteen human cadaveric knees were tested. Windows were made in the joint capsule and strain gauges inserted into the anterior, middle and posterior sections of the medial meniscus. The knees were then loaded to three times body weight at speeds of 50 and 500 mm/min, with the knee joint at 0 degrees and 30 degrees of flexion. The tests were repeated following the creation of a longitudinal or a radial tear in the meniscus. RESULTS: The intact menisci showed significantly less strain in the posterior section compared to the anterior and middle sections (P < 0.003, with strains of 1.54%, 2.86% and 2.65% respectively). With a longitudinal tear this pattern changed with strains decreasing anteriorly and increasing posteriorly. There were also significant differences at different angles of knee joint flexion not seen in the intact meniscus. 50% radial tears reduced the strains anteriorly whilst a complete radial tear completely defunctioned the meniscus. CONCLUSIONS: This study has shown that there are significantly different hoop strains produced in different sections of the medial meniscus under load and the patterns of strain distribution are disturbed by meniscal tears. RELEVANCE: These results provide important data for mathematical models which must include non-uniform behaviour. They also have implications for the surgical management of torn menisci. Undamaged portions should be preserved and the integrity of the circumferential fibres maintained to ensure the menisci retain a load bearing capability. PMID- 11415636 TI - Observations on the F-Scan in-shoe pressure measuring system. AB - OBJECTIVE: To evaluate the physical characteristics (durability, calibration, creep and hysteresis) and the accuracy and repeatability of the F-Scan in-shoe sensor. DESIGN: Observations of the system in clinical use combined with material bench-test experiments. BACKGROUND: The F-Scan system is a commercially produced in-shoe force/pressure measurement system. Whilst useful clinical studies have been reported there is little information on the performance characteristics of the system. METHODS: Output mass error following calibration was directly measured from the F-Scan system. A jig-mounted force meter tested groups of individual sensing units (4 @ 1 cm(-2)) for within and between sensor accuracy in full size and adjusted sensors. An Instron servohydraulic materials testing unit was used to evaluate creep, hysteresis and the repeatability of output forces over repeated loading cycles for full size and cut sensors. RESULTS: The F-Scan system lacks durability and suffers significant calibration error. Creep (19%) and hysteresis (21%) properties were poor. Within and between sensor variability in output was demonstrated and overall repeatability was poor. Adjusting the sensor size adversely affects output. CONCLUSIONS: Our findings suggest the F Scan system has a limited capability for absolute accuracy but could still be used for quantitative studies provided its limitations are noted. RELEVANCE: These findings may help F-Scan users evaluate the relevance of clinical data and refine measurement protocols in the context of their own field of work. PMID- 11415637 TI - Thoracolumbar disc degeneration in young fast bowlers in cricket: a follow-up study. AB - OBJECTIVE: To determine the progression of thoracolumbar disc degeneration in young fast bowlers in cricket. DESIGN: Prospective fast bowling technique and MRI follow-up study. BACKGROUND: Previous studies on high-performance young fast bowlers have found that lumbar spine pathology was related to the mixed bowling technique. METHODS: Nineteen young male fast bowlers (mean age 13.6 years) underwent MRI scans to detect the presence of intervertebral disc abnormalities. Subjects were also filmed laterally (200 Hz) and from directly above (100 Hz) whilst bowling two maximum velocity deliveries (session 1). Subjects were tested using an identical methodology 2.7 years later (session 2). RESULTS: At session 1, the incidence of thoracolumbar disc degeneration was 21%; however, at session 2, the incidence significantly (P = 0.008) increased to 58%. Furthermore the increase in the incidence of back pain between session 1 and session 2 was also significant (P = 0.002). The progression of disc degeneration was found to be significantly (P = 0.015) related to the group of fast bowlers who utilized the mixed technique during both session 1 and 2 when compared to those who used this technique during one session only. CONCLUSIONS: Thoracolumbar disc degeneration and back pain increase significantly during the time period examined in this study. Further, bowlers who utilize the mixed bowling technique stand a greater chance of developing degenerative changes of the spine. PMID- 11415638 TI - An in vivo study of the primary and coupled rotations of the thoracic spine. AB - OBJECTIVE: To provide preliminary data on three-dimensional thoracic spine kinematics measured in vivo. DESIGN: This study measured the three planes of thoracic spine motion in normal subjects using an external measuring device. BACKGROUND: Few studies have investigated the primary and associated coupled rotations in the thoracic spine in vivo. Most knowledge of motion characteristics comes from in vitro studies which have limitations. There is a lack of agreement on the patterns of thoracic coupled motion especially that between lateral flexion and axial rotation. METHODS: Thoracic motion was examined in 60 normal subjects (30 males, 30 females) aged 18-24 years. The primary and coupled rotations of the thoracic regions T(1-4), T(4-8), T(8-12) were measured using a 3 SPACE Fastrak system. RESULTS: The three thoracic regions displayed the characteristic variations in range and distribution of primary rotations previously described. The pattern of coupled motion varied between subjects but an ipsilateral pattern predominated between lateral flexion and axial rotation in the middle and lower thoracic regions while the upper thoracic region was found to exhibit either a contralateral or ipsilateral pattern. Gender did not influence results. CONCLUSIONS: The pattern of coupled motion in the thoracic spine demonstrated some variability between subjects in vivo. Lateral flexion and axial rotation were strongly coupled with overall, their relationship being predominantly ipsilateral. PMID- 11415639 TI - Reproducibility of lumbar paraspinal surface electromyogram power spectra. AB - OBJECTIVE: To establish the reproducibility of surface recordings of electromyograms from lumbar erector spinae muscles. DESIGN: Repeated measurements in 10 male volunteers under controlled conditions. METHODS: While isometric load was held constant at (2/3) and (1/3) of maximum voluntary contraction, surface electromyograms were recorded from the 4th/5th lumbar interspace on three occasions separated by at least a day. Fresh surface electrodes were applied on each test occasion. The raw electromyographic signal was filtered and digitized and the signal processed by fast Fourier transformation to give median frequency and total amplitude spectra. Linear regression lines of median frequency and amplitude against time were calculated. The 'spectral halfwidth" was defined as the full width at half maximum of the composite amplitude spectrum. Reproducibility was assessed by calculation of the intraclass correlation coefficient. RESULTS: Reproducibility of initial median frequency and spectral halfwidth were found to be excellent at both (2/3) (ICCC 0.91 and 0.88) and (1/3) (ICCC 0.84 and 0.83) maximum voluntary contraction. Initial power and regression slopes of power and median frequency were not reproducible. CONCLUSIONS: The reproducibility of two electromyographic variables, the initial median frequency and the spectral halfwidth, were found to be satisfactory. PMID- 11415640 TI - Clinical and biomechanical aspects of external fixation of the pelvis. AB - OBJECTIVE: The aim was to evaluate the mechanical stability of several traditional and modern external fixators in unstable pelvic ring disruption. DESIGN: In a laboratory study external and internal fixation techniques were tested in seven fresh and five embalmed human pelves with a disruption of the pubic symphysis and one sacroiliac joint (type C1.2 injury according to the Tile AO classification). BACKGROUND: Stability provided by external fixation depends upon many factors, with the residual pelvic stability being the most important. METHODS: Simulating a single-leg stance, the load was applied quasi-statically to the acetabulum of the unstable hemipelvis. Device failure was defined as displacement >10 mm either at the symphysis pubis or the sacroiliac joint. RESULTS: The frame with the highest failure load (fresh versus embalmed specimens) was the Egbers configuration with the AO fixator (analysis of variance; P < 0.05). Failure was noted at 114.9 N versus 129.5 N. Augmentation of the Mono-Tube by additional internal posterior osteosynthesis gave the following results: sacral bars 325.4 N versus 217.8 N, plate fixation 294.3 N versus 215.8 N, lag screws 338.4 N versus 215.8 N. Failure loads of hybrid fixation of the Orthofix were as follows: sacral bars 257.9 N versus 213.9 N, plate fixation 333.5 N versus 245.3 N, lag screws 397.3 N versus 280.6 N. The differences between the two fixators were not statistically significant. CONCLUSIONS: No single external frame provided sufficient stability. The addition of a posterior internal fixation significantly increased failure loads and controlled the weight bearing pelvic elements. PMID- 11415641 TI - In vivo validation of in vitro testing of hallucal flexor mechanics. AB - OBJECTIVE: The purpose of this study was to validate an in vitro method of testing first metatarsophalangeal joint mechanics. DESIGN: We compared the orientation of the resultant moment of hallucal plantarflexion in cadaveric specimens undergoing loading of the flexor hallucis longus to the plane of the active great toe plantarflexion moment in normal volunteers. BACKGROUND: Frontal plane deviation of hallucal plantarflexion is a fundamental component of the altered biomechanics in hallux valgus. METHODS: The cadaveric model utilized dynamic loading of the flexor hallucis longus and static loading of the remaining muscles of the great toe. The in vivo method involved measurement of force output with isometric hallucal plantarflexion in the standing position. The medial lateral and superior-inferior forces as well as the angle of the resultant moment in the clinically relevant frontal plane for both groups were quantified. RESULTS: We found no significant difference between the frontal plane forces of each group. The directional orientations in the frontal plane also displayed a marked similarity, with an average lateral deviation of 0.63 degrees (P = 0.99) for each group. CONCLUSIONS: The lack of a significant difference between the cadaveric and clinical methods of finding the resultant hallucal moment indicates that the cadaveric method accurately simulates in vivo internal flexor muscular loading of the great toe. PMID- 11415642 TI - Maximum voluntary weight-bearing by the affected and unaffected legs in standing following stroke. AB - OBJECTIVE: To compare stroke patients to control subjects for ability to transfer body weight onto the affected and unaffected leg in standing; to investigate intra-session reliability. DESIGN: Comparative clinical study conducted within a single session. BACKGROUND: There is a paucity of quantitative data about maximum voluntary weight-bearing in patients during rehabilitation following stroke. METHODS: A Kistler force platform was used to quantify maximum amount of body weight transferred to a single limb in the lateral and forward directions during weight-shifting. Twelve control subjects matched by gender and age (median 64 years) were compared to 12 inpatient stroke patients after a median of 37 days post-onset. RESULTS: The median score for control subjects was approximately 95% of body weight to each leg in both directions. In contrast, stroke patients transferred less body weight (P<0.01) to the affected leg (65.5% lateral; 54.9% forward) and also to the unaffected leg (85.0% lateral; 80.1% forward). For the stroke patients, transfer of body weight was more challenging in the forward direction than the lateral direction on the affected leg (P<0.05). Relative to individual differences in the stroke group, error due to the repeated measurement process was low. CONCLUSION: The testing procedure was found to discriminate between stroke patients and control patients and had high retest reliability within a single session. PMID- 11415643 TI - Failure of stimulated skeletal muscle mainly contributed by passive force: an in vivo rabbit model. AB - OBJECTIVE: To evaluate the role of active and passive muscle forces in the failure mechanism of stimulated muscle. DESIGN: An in vivo rabbit model. BACKGROUND: Eccentric contractions can result in a greater incidence of muscle injury. However, the relative role of the active and passive muscle force in the failure mechanism of the activated muscle is not well elucidated. METHODS: After anaesthesia, New Zealand white rabbits were fixed in a frame on a materials testing machine. The triceps surae muscle-tendon units were passively stretched to rupture with our without continuous nerve stimulation. The force and muscle length were simultaneously recorded. Active muscle force, passive muscle force, and ratio of the active to passive muscle were calculated and depicted against strain. RESULTS: The results showed that the mean maximal passive force of triceps surae muscle was 293.1 N at a strain of 38%. The mean peak active muscle force was 21.5 N at a strain of 21%. The ratio of active to passive muscle force reached its peak first, followed by the active muscle force, and then the passive muscle force. The ratio of active to passive muscle force at the peak total force was only 3.3%. CONCLUSIONS: The stimulated muscle can exert its maximal response at extreme physiological extension. Injury of the stimulated muscle is caused mainly by passive muscle force. PMID- 11415644 TI - Combined models of ultrasound velocity and attenuation for predicting trabecular bone strength and mineral density. AB - Three quantitative ultrasound parameters and their combinations were examined as possible predictors of the bone mineral density and compressive strength of human cancellous bone. Seventy-three trabecular bone cubes obtained from female cadaveric tibiae were measured by quantitative computed tomography. Measurements of the ultrasound velocity, ultrasound attenuation at 0.5 MHz and broad-band ultrasound attenuation were made in three orthogonal anatomical directions. The specimens were then tested to failure in compression, 38 in the anterior/posterior direction and 35 in the superior/inferior direction. The apparent density of all specimens was also found. Linear regressions showed that bone mineral density and strength were significantly correlated with each ultrasonic parameter in each direction (P < 0.05). Linear combinatinations of two ultrasound parameters were significantly better predictors than single parameters, although the optimum pair of parameters differed with direction. Three parameter models did not further improve predictability. PMID- 11415645 TI - Bilateral femoral rotations measured during walking: a new parameter to summarize and describe individual gait. AB - The aim of this study was to develop and assess a method of analysis of femoral rotations to describe and summarize an individual's gait. Twelve normal subjects underwent walking trials at both natural and slow speeds, controlled by a metronome. The three-dimensional rotations of each femur were measured, simultaneously, during walking using a 3SPACE Tracker. The phase lags between the three rotations for each leg, obtained by cross-correlations, provided the primary data. The device was found to be reliable and gave significant test- retest repeatability. The analyses provided summary parameters describing mathematically the patterns of rotations and showed there was repeatability within and between sessions. A strong systematic subject effect in all analyses indicated that these parameters provide a highly specific description of how an individual walks. PMID- 11415646 TI - Appropriateness of plane pressure-sensitive film calibration for contact stress measurements in articular joints. AB - OBJECTIVE: To examine how accurately calibration stains from pressure-sensitive film samples compressed between perfectly flat surfaces represent contact pressures measured with the film inserted between curved joint surfaces. DESIGN: Experimental study. BACKGROUND: Contact pressure distributions and areas in joints are often quantified using pressure-sensitive film inserted between the articular surfaces. Absolute pressure values are recovered from the film through a calibration procedure. Since the calibration is typically performed in a different environment from the experimental measurements, a systematic error in the pressure magnitudes may be introduced. METHODS: Calibration stains were produced with an Instron machine using a flat and a slightly curved indenter. Calibration curves were fitted to the raw data of both indenters using a model representing the functional characteristics of the pressure-sensitive film. RESULTS: No systematic differences between the calibration curves from the flat and the curved indenter were observed. CONCLUSIONS: It appears that calibration with the film in a flat configuration is adequate for contact pressure measurements between moderately curved articular surfaces. PMID- 11415647 TI - The reliability of postural sway measures using the 3SPACE Tracker. AB - OBJECTIVE: To investigate the reliability of three-dimensional postural control sway measures, in a normal population, using an electromagnetic device. DESIGN: A repeated measures design was used within and between sessions. BACKGROUND: Electromagnetic measurement has been proposed as a method of measuring postural sway; however, reliability within a normal population has not been shown. METHOD: An electromagnetic device, the 3SPACE Tracker, measured postural sway at the pelvis using sway path length and displacements in the anterior-posterior, medial lateral and vertical direction. Ten subjects performed a postural control task involving two trials for six different sensory conditions, on two separate occasions, two weeks apart. RESULTS: The reliability of the technique was demonstrated by the repeatability of results for individual subjects within and between sessions. Individual subjects and sensory conditions were discriminated. Sway path length was the most consistent of the measures used. CONCLUSIONS: Three dimensional measures of postural sway are reliable and discriminatory in a normal population. PMID- 11415648 TI - The accumulation of fatigue microdamage in human cortical bone of two different ages in vitro. AB - OBJECTIVE: To analyse the development of damage during fatigue cycling of human bone. DESIGN: Changes in compliance and the cycles to failure were monitored in cortical bone samples subjected to oscillating stress in vitro. BACKGROUND: Previous studies produced mainly the relationship between the applied stress and the final cycles to failure (sigma-N(f)plots). However, cyclic stressing increases the compliance of the bone continuously, and causes a progressive mechanical/structural degradation. Recording this accumulation of damage allows one to know how close bone is to the point of failure; more importantly, it allows a more comprehensive modelling of fatigue processes in cortical bone. METHODS: The occurrence of material damage was continuously monitored during the tests. The 20 specimens came from two female subjects, 27 and 56 years old. The range of the cyclic stresses was 58-130 MPa. RESULTS: The damage was quantified with a graphical and an empirical/numerical method, and we have also microscopically observed the generation of internal microcracks. The range of cycles to failure was from 1 to 210,000. CONCLUSIONS: It was observed that (i) the older tissue showed a lower fatigue strength than the younger one, (ii) both tissues sustained similar damage levels prior to failure, and (iii) they both showed a continuous accumulation of damage during the tests, the course of which depended on the level of stress. PMID- 11415649 TI - Assessment of scoliotic deformity from back shape asymmetry using an improved mathematical model. AB - OBJECTIVE: The objective of the study was to improve with respect to accuracy and smoothness the three-dimensional model of the spinal midline which has been calculated from rasterstereographic back surface data. DESIGN: A new mathematical model using frequency-modulated sine curves has been applied to existing rasterstereographic and radiographic data in an in vivo study. BACKGROUND: Analysis of back shape (measured by rasterstereography) enables a three dimensional model of the spinal midline to be calculated. The model is based on the so-called symmetry line which coincides approximately with the line of the spinous processes. In addition the surface rotation on the symmetry line is taken as an estimate for vertebral axial rotation in scoliosis. METHODS: Four hundred and seventy-eight pairs of radiographs and rasterstereographs of scoliotic patients have been evaluated and compared using the new model. The radiographs were digitized manually, delivering curves of lateral deviation and vertebral rotation. Analysis of back shape from the rasterstereographs delivers a three dimensional model of the spinal midline, the frontal projection of which is to be compared with the radiographic curve. Likewise, vertebral rotation and surface rotation are compared. RESULTS: The use of frequency-modulated sine curves improved the results as compared to the existing procedure with respect to prediction accuracy of lateral deviation (typically from sigma(x) = 4.6 mm to sigma(x) = 4.0 mm, P < 10(-10)). The prediction of axial rotation was not significantly improved (sigma(varrho) thick approximate 3.8 degrees ). In addition, the smoothness of the curves is considerably improved. The prediction accuracy proved to be independent of the scoliosis type as represented by apex height (r = -0.09 ellipsis + 0.03). CONCLUSION: The use of frequency-modulated sine curves improves significantly the accuracy and smoothness of the spinal model curves. PMID- 11415650 TI - Quantitative assessment of the control capability of the trunk muscles during oscillatory bending motion under a new experimental protocol. AB - OBJECTIVE: A new quantitative technique for measuring the trunk control capability/coordination was to be developed in this study. DESIGN: Fitts' experimental paradigm was employed to quantify the information processing capacity (bits/s) of the trunk as well as dynamic motor performance such as velocity and acceleration during flexion and extension. BACKGROUND: The quantification of functional capability of the trunk such as range of motion, strength, and endurance have been used to evaluate low back pain patient. Especially, dynamic trunk motion during flexion and extension has been studied not only to quantify the severity of the low-back impairment but to classify patients. METHOD: A lumbar motion monitor was used to record the time series of range of motion (RoM) and compute the velocity and acceleration of the trunk motion. Twenty male subjects without any back pain in the past 6 months and previous history of back injury participated. Each subject performed 22 controlled flexion/extension at predetermined RoMs as well as one ballistic trunk flexion/extension at a self-selected RoM. RESULTS: The information processing capacity of the trunk among healthy subjects was found to have a mean of 4.23 (SD 1.43) bits/s based on Fitts' law. Also, the velocity of dynamic trunk motion was measured with a considerable reduction in intersubject variability when the RoMs were controlled. A short but still accurate experimental protocol was suggested via a series of statistical analyses to provide an objective and easy-to-use method to evaluate the functional capacity of low-back pain patients. PMID- 11415651 TI - EMG activities of the quadratus lumborum and erector spinae muscles during flexion-relaxation and other motor tasks. AB - OBJECTIVE: The aim of this study was to provide new information on the myoelectrical activation of the quadratus lumborum, the deep lateral and the superficial medial lumbar erector spinae, the psoas, and the iliacus muscles in various motor tasks. DESIGN: An intramuscular electromyographic study was performed. BACKGROUND: The contribution of individual deep trunk muscles to the stability of the lumbar spine is relatively unknown in different tasks, including the flexion-relaxation phenomenon. METHODS: Seven healthy subjects participated. Fine-wire electrodes were inserted with a needle guided by ultrasound. RESULTS: The highest activity observed for quadratus lumborum and deep lateral erector spinae occurred in ipsilateral trunk flexion in a side-lying position and for superficial medial erector spinae during bilateral leg lift in a prone position. Quadratus lumborum and deep lateral erector spinae were activated when the flexion-relaxation phenomenon was present for superficial medial erector spinae, i.e. when its activity ceased in the latter part of full forward flexion of the trunk, held relaxed and kyphotic. CONCLUSIONS: In general, the activation of the investigated muscles showed a high degree of task specificity, where activation of a certain muscle was not always predictable from its anatomical arrangement and mechanical advantage. PMID- 11415652 TI - Passive properties of the forearm musculature with reference to hand and finger postures. AB - OBJECTIVE: To quantify individual forearm muscle passive forces and evaluate their impact on hand function. DESIGN: The passive force-length properties of the 24 extrinsic hand and wrist muscles were determined in five fresh frozen cadaver arms. BACKGROUND: Muscle force production is a summation of the active and passive force components. The passive properties of the extrinsic finger musculature and wrist musculature appear to strongly affect both hand posture and hand movement. METHODS: The passive force-length properties of extrinsic hand and wrist muscles were determined by applying a slow, continuous extension to each muscle and recording the resulting tension. Each force-length curve was fit using exponential regression and were related to specific joint rotations and seven hand postures by calculating the muscle excursions for those postures. RESULTS: The exponential passive force-length relationship explained over 97% of the experimental variance. The largest passive forces were elicited in the digital extensors in grips involving large flexion angles such as tip pinch, key pinch, and a briefcase grip. CONCLUSIONS: The passive properties of the extrinsic finger musculature and wrist musculature affect both hand posture and movement especially in postures with flexed wrist and fingers. PMID- 11415653 TI - Microcirculation in the footsole as a function of mechanical pressure. AB - OBJECTIVE: In this study an experimental set-up for measuring skin microvascular responses of the footsole to changes in externally applied pressure was analysed. DESIGN: A clinical study. Skin microvascular blood flow was measured in healthy volunteers, during and after external mechanical pressure of different magnitudes. BACKGROUND: During standing and walking the footsole is commonly exposed to high static and dynamic mechanical pressure, resulting in changes in the microcirculation of the footsole. In diabetic patients a disturbed interaction between externally applied pressure and skin microvascular response seems to be involved in the development of a foot ulcer. METHODS: Eleven volunteers participated in the study. Static loads were applied to the heel part of the footsole with the person in a supine position. Contact pressure and skin blood flux, based on the laser Doppler technique, were simultaneously monitored. The pressure used was varied in five discrete steps between 10 and 160 kPa and applied during a period of 5 min each. The microcirculation was measured during as well as after pressure loading. RESULTS: Pressures of 40 kPa and higher do stop the blood flow in the skin microcirculation. Releasing the applied pressure resulted in a hyperaemic response. This response appears to increase in amplitude at increasing pressures up to 800% of the baseline laser Doppler fluxmetry level. Beyond a pressure level of 80 kPa the hyperaemic response seems not to be influenced by the pressure level. The time needed to achieve the maximal laser Doppler fluxmetry level decreased when the pressure was raised from 10 to 80 kPa, but increased again when higher pressures were applied (P = 0.051). An intraindividual variation of 11-50% was observed for the parameters describing the blood flux before, during, and after pressure application. CONCLUSION: Simultaneously measuring changes in contact pressure and laser Doppler flux of the footsole is a useful method to study the interaction of external mechanical pressure and skin microvascular reactions. Pressures above 40 kPa stop skin microvascular blood flow. Releasing the applied pressure results in a hyperaemic response, which increases when the applied pressure increases from 40 to 80 kPa. Higher pressures do not influence the amplitude in skin microvascular response, but result in a longer delay to maximal hyperaemia. PMID- 11415654 TI - Intratester and intertester reproducibility of the lumbar motion monitor as a measure of range, velocity and acceleration of the thoracolumbar spine. AB - OBJECTIVE: The purpose of this study was to examine the intra- and intertester reproducibility of the Lumbar Motion Monitor (LMM) as a measure of thoracolumbar range of motion (RoM), velocity and acceleration. DESIGN: The study was a clinical trial using asymptomatic subjects. BACKGROUND: The LMM is a promising device from a clinical and research perspective, though the reproducibility of it's measurements has yet to be fully determined on subjects under free motion conditions. METHODS: For intratester reproducibility, 15 subjects were required to move as far and as fast as they could in 8 s through flexion, side flexion, and rotation. They were tested on three occasions with 48 h separating tests. For intertester reproducibility, 10 subjects were tested by two examiners, and were required to move as far and as fast as they could in 8 s. RESULTS: Intratester reproducibility coefficients ranged between 0.82 and 0.87 for RoM, 0.61 and 0.87 for velocity, and 0.46 and 0.72 for acceleration. Rotation had the poorest reproducibility in each instance. Intertester reproducibility ranged between 0.93 and 0.98. CONCLUSIONS: The reproducibility of the LMM is suitably high for RoM and velocity for the device to be used for evaluation in a clinical and research setting. PMID- 11415655 TI - Artificial intelligence analysis of paraspinal power spectra. AB - OBJECTIVE: As an aid to discrimination of sufferers with back pain an artificial intelligence neural network was constructed to differentiate paraspinal power spectra. DESIGN: Clinical investigation using surface electromyography. METHOD: The surface electromyogram power spectra from 60 subjects, 33 non-back-pain sufferers and 27 chronic back pain sufferers were used to construct a back propagation neural network that was then tested. Subjects were placed on a test frame in 30 degrees of lumbar forward flexion. An isometric load of two-thirds maximum voluntary contraction was held constant for 30 s whilst surface electromyograms were recorded at the level of the L(4-5). Paraspinal power spectra were calculated and loaded into the input layer of a three-layer back propagation network. The neural network classified the spectra into normal or back pain type. RESULTS: The back propagation neural was shown to have satisfactory convergence with a specificity of 79% and a sensitivity of 80%. CONCLUSIONS: Artificial intelligence neural networks appear to be a useful method of differentiating paraspinal power spectra in back-pain sufferers. PMID- 11415656 TI - The static and dynamic behaviour of tibial fractures due to unlocking external fixators. AB - OBJECTIVE: To determine how the mechanical environment of a tibial fracture is influenced by unlocking an external fixator frame. DESIGN: A clinical study examined 10 consecutive patients fixed with the Orthofix DAF. BACKGROUND: It has been claimed that the healing of diaphyseal tibial fractures is assisted by unlocking external fixators to allow free axial movement, but the influence on the mechanical environment at the fracture has not been established. METHODS: A transducer attached to bone screws measured dynamic interfragmentary displacement during walking both before and immediately after unlocking the fixator at 6 weeks in 10 subjects. Four subjects were monitored over the first hour after unlocking to measure interfragmentary gap shortening. RESULTS: Mean peak amplitudes of cyclical axial and angular displacement before unlocking were 0.46 mm (SD 0.27) and 0.37 degrees (SD 0.30), and after were 0.42 mm (SD 0.19) and 0.34 degrees (SD 0.28). Mean peak torsional and transverse shear displacements were 0.21 degrees (SD 0.11) and 0.30 mm (SD 0.17) before unlocking, and after were 0.42 degrees (SD 0.39) and 0.51 mm (SD 0.60). Gaps shortened permanently by unrecoverable axial translations of between 0.2 and 1.4 mm; the mean was 0.69 mm. CONCLUSIONS: Unlocking was found more often to reduce both axial and angular motion, but to increase shear. Overall, this may reduce maximum longitudinal strains in the external callus. The reduced motion may arise from gap shortening. PMID- 11415657 TI - Load sharing in augmented anterior cruciate ligament repair: a mathematical analysis based on in vitro measurements. AB - OBJECTIVE: To quantify the stress on a reinserted anterior cruciate ligament (ACL) we studied the load sharing between the ACL and a 3.7 mm polyethylene terephthalate (PET) band in 10 knees of fresh human cadavers. DESIGN: The load sharing between the Marshall sutures-ACL complex and the PET band and between the ACL and the PET band was calculated by means of a mathematical model. BACKGROUND: Augmentation of a reinserted ACL with a synthetic band is an accepted treatment for a ruptured ACL. METHODS: After transsecting the ACL at its femoral origin and reinserting it with four sutures using the Marshall technique we augmented it with the PET band. The augmentation device was inserted by the through-the condyle (TTC) procedure and attached without preload to the femoral and the tibial condyle with 4 mm staples. Then the length of the ACL, the lengths of the Marshall sutures, the partial lengths and the angles between the different directions, and the adherence-friction force of the PET band were measured. RESULTS: In the beginning the Marshall sutures-ACL complex takes over 40% of an externally applied load and the PET band 60%. After the hypothetical ACL healing phase the PET band takes over 27% and the ACL 73% of the load. CONCLUSIONS: An external force of 40 N acting on the knee will therefore lead to an elongation of the Marshall sutures-ACL complex and result in movement of the proximal end of the ACL away from the femoral condyle of at least 0.5 mm, which is highly undesirable in the early postoperative healing phase. PMID- 11415658 TI - Passive knee muscle moment arms measured in vivo with MRI. AB - OBJECTIVE: To determine moment arm lengths from seven knee muscles and the patellar tendon. The knee muscles were the biceps femoris, semitendinosus, semimembranosus, gracilis, sartorius, and the lateral and medial gastrocnemius muscles. DESIGN: The moment arms were calculated based on MRI measurements. BACKGROUND: Moment arm lengths of different muscles with respect to the joint centre of rotation (CR) or the centre of the contact point between joint surfaces are necessary basic data for biomechanical models predicting joint load. METHODS: Ten male and seven female subjects participated. Using a 1.5 Tesla magnetic resonance imaging system, 3-dimensional coordinates of relevant points were recorded from a 3-D volume reconstruction of the right knee at knee flexion angles of 0, 30 and 60 degrees. Muscular moment arms were calculated in both the sagittal and frontal planes. The recordings were all made during passive mode, which means that no muscular contraction was performed. RESULTS: All muscles except the lateral gastrocnemius showed statistically significant differences (P<0.05) of moment arm lengths between gender in the frontal plane. All muscles except biceps femoris and sartorius showed significant differences (P<0.05) of moment arm lengths between gender in the sagittal plane. Most muscles also showed a linear or quadratic trend of changing moment arms with varying knee angle. CONCLUSIONS: Our results indicate that for most biomechanical analyses involving knee muscles, gender- and angle-specific moment arms should be used. PMID- 11415659 TI - Diagnosis of knee instability using sonography. AB - OBJECTIVE: A sonographic method has been developed to diagnose knee joint instabilities and to gain exactly quantifiable and reproducible measurements. DESIGN: Based on biomechanical considerations, a new positioning device to analyse knee instabilities sonographically has been developed and tested in in vivo studies. BACKGROUND: Common instrumental testing techniques are difficult to reproduce and show a lack of reliability. METHOD: Using sonography the cortical line of the femur and tibia are observed. At the same time a translation of these two partners at the knee joint is provoked in a defined position using the special holding device and under a defined amount of stress. Using this method 142 healthy people, placed in different age groups, were tested to establish standard values for the physiological knee joint translation. A prearthroscopic stability analysis was performed on 101 patients. RESULTS: This study proves that the special device developed, together with the technique used, provides reliable and exact measurements for knee instability in different translation directions. In healthy subjects the lateral compartment always showed a larger translation than the medial compartment. A gradual increase in translation was determined in the age groups over 30 years. A significant difference of 0.0001 was shown between knee stability and instabilities. Moreover, a further differentiation between partial and total ACL ruptures and chronic instabilities was determined. This provides an important advantage not offered by usual instrumental testing techniques. CONCLUSIONS: The ability to diagnose even a very low extent of instability and to differentiate between different kinds of instabilities enables exact controls to be carried out for both conservative and operative treatment. PMID- 11415660 TI - Precision measurement of segmental motion from flexion-extension radiographs of the lumbar spine. AB - OBJECTIVE: To measure sagittal plane motion of lumbar vertebrae from lateral radiographic views. Previously identified factors of imprecision such as distortion in central projection, off-centre position, axial rotation, and lateral tilt of the spine were compensated. STUDY DESIGN: This study presents a new protocol to measure sagittal plane rotational and translational motion from lateral flexion-extension radiographs of the lumbar spine. BACKGROUND: Conventional methods to determine sagittal plane rotation and translation are prone to error from the distortional effects of the divergence of the radiographic beam and the measurement error inherent in constructing tangents to the contours of the vertebral body. High precision is attained by roentgen stereophotogrammetric methods, but because of their invasive nature they can be applied only in exceptional cases. Agreement has been reached only in that measurement of sagittal plane motion from lumbar spine flexion-extension radiographs is inaccurate. Normal patterns of sagittal plane motion and the definition of what is an abnormal flexion-extension radiograph have not been settled. METHOD: Starting from an analysis of vertebral contours in the lateral view, geometric measures are identified which are virtually independent of distortion, axial rotation or lateral tilt. Applying a new protocol based on those geometric measures, the pattern of translational and rotational motion was determined from flexion-extension radiographs of 61 symptom-free, adult subjects. Measurement errors were quantified in a specimen experiment; a reproducibility study quantified inter- and intraobserver errors. RESULTS: Magnitude and sign of 'translation per degree of rotation' determined from a cohort of 61 adult subjects were very uniform for all levels of the lumbar spine. An auxiliary study evaluating a cohort of 10 healthy subjects where flexion-extension radiographs had been taken standing and side-lying showed no dependence of the rotation/translation pattern on posture. The error study demonstrated errors in angle ranging between 0.7 and 1.6 degrees and errors in displacement ranging between 1.2% and 2.4% of vertebral depth (the largest errors occurring at the L(5)/S(1) segment). Intra- and interobserver tests showed no or only negligibly small bias and an SD virtually equal to the measurement error multiplied by radical2. The relation of displacement to angle observed in the normal cohort can be used in individual cases to predict translational motion depending on the rotation actually performed. A comparison of the predicted translation (determined from normal controls) and the value actually measured allows translational hypo-, normal, or hypermobility to be quantified. Examples illustrate application of the new method in cases of normal, hypo-, and hypermobility and in the case of an instrumented spine. CONCLUSIONS: The results of this study show that precision of the measurement of rotational and translational motion can be considerably enhanced by making allowance for radiographic distortional effects and by minimizing subjective influence in the measurement procedure. PMID- 11415661 TI - Can the computerized physical examination differentiate normal subjects from abnormal subjects with benign mechanical low back pain? AB - Inconsistencies among physicians in the evaluation of benign low back conditions make standardization desirable. A computerized physical examination device was used to evaluate low back pain patients and compare their results with a normative database obtained from a selection of healthy subjects. A high resolution motion analysis system tracked the movement of skin markers placed on the midline and pelvis. Surface electromyography electrodes placed above L(5) collected data from multifidus. From the kinematics of skin markers during flexion extension with lifts up to 32 kg, and lateral bending with lifts up to 4.6 kg, the following parameters were estimated: lumbosacral angle and elongation, contribution of each lumbar segment to the lordosis reduction, relative pelvic/spine motion and trunk velocity. First, the average normal value for each estimated parameter was determined using 40 normal subjects. For each subject, the difference between his parameter and the normal was processed by an expert system generating a normality index varying from zero (perfect abnormal) to one (perfect normal). To develop the expert system's rules, a preliminary group of 20 very abnormal subjects were used, such that the normality index separated them from the normals. For validation, a set of 29 WCB sprain patients and another set of 42 discogram positive were selected. Each subject was tested and his computerized normality index calculated without any clinicians' input. The computerized normality index was compared with the clinicians' evaluation which was taken to be the gold standard. The Receiver Operating Characteristic technique was used to quantify the discrepancies. Results show that the expert system can detect clinically abnormal subjects with accuracy (sensitivity 83-91% and specificity >/=90%) while providing quantitative information on workers' functional capacities. PMID- 11415662 TI - A non-invasive protocol for the determination of lumbar spine mobility. AB - OBJECTIVE: This article presents a non-invasive protocol for the determination of lumbar spine mobility and compares intersegmental lumbar spinal mobility data from 45 subjects using both radiographic (invasive) and videographic (non invasive) techniques. DESIGN: Non-linear mathematical models were developed to transfer skin marker coordinates into corresponding vertebral body positions. BACKGROUND: To relate skin surface motion with that of the underlying vertebrae and to enhance the reliability of using the non-invasive technique relied on a model of transformation. METHODS: This set of models was developed by taking the L(4) skinfold, the L(1)-S(1) skin distraction, and the non-linear difference between the lumbar spinal profile and the skin profile into consideration. Based on the digitized and model transformed data, the intersegmental joint angles (ISA) and intersegmental joint mobilities (ISM) were calculated and compared. RESULTS: In this study the non-invasive and invasive coordinate differences ranged from 0.485 to 1.099 cm; the mean ISA differences ranged from 0.29 to 2.08 degrees the mean ISM differences ranged from 0.62 to 1.88 degrees. CONCLUSIONS: With these small differences, the quality of the non-invasive protocol and the application of the transformation model were justified sufficient. PMID- 11415663 TI - Validation of a 3D optoelectronic motion analysis system for the wrist joint. AB - OBJECTIVE: A study was undertaken to determine the experimental accuracy of a non invasive optoelectonic 3-dimensional tracking system in assessing wrist joint motion. DESIGN: This was an in vivo experimental study involving volunteer subjects performing prescribed wrist motions. BACKGROUND: Current clinical practice does not include routine kinematic analysis for evaluating arthritic disease state, although motion disorders are common. METHODS: Surface markers were applied to 24 subjects assigned two hand postures in a test-retest factorial design for the expected range of motion. The marker positions were measured optoelectronically and using calibrated stereoradiography, to determine the positions of the surface markers and of key bone landmarks. Alignment and motion were compared for the three measurement techniques. Standard kinematic analyses were performed to extract Euler angles and equivalent screw displacement axes for paired postures. RESULTS: The three measurement techniques were highly correlated for wrist flexion-extension. Uncertainties were less than 6 degrees, similar to uncertainties from bone landmark identification errors when implanted markers cannot be used. Measures of motion exhibited higher correlations than those for alignment. Equivalent screw displacement axis orientations had poor intraclass correlations, reflecting sensitivity to coordinate system definitions. CONCLUSIONS: For motion analysis in the wrist in vivo, a non-invasive optoelectronic measurement system is as accurate as stereoradiographic analysis of bone segments. PMID- 11415664 TI - Angular movements of the lumbar spine and pelvis can be reliably measured after 4 minutes of treadmill walking. AB - OBJECTIVE: To determine the familiarization period required to obtain consistent measurements of the angular movements of the lumbar spine and pelvis during treadmill walking. DESIGN: An in vivo study with repeated measures every 2 min over 10 min. BACKGROUND: Walking on a treadmill can initially be an unfamiliar experience. No data were available to indicate the length of time required for treadmill familiarization prior to taking measurements of the angular movements of the lumbar spine and pelvis. The familiarization period has implications for the use of this technique in clinical testing. METHODS: The angular movements of the lumbar spine and pelvis were examined by thePEAK 3D motion measurement system in 16 untrained, normal subjects walking for 10 min on a treadmill at either self selected or 60% of self-selected speed. RESULTS: The reliability of the angular measurements of the spine and pelvis were all greater than 0.83 (ICC) after 4 min of treadmill walking and did not appear to increase after 4 min. No changes in the amplitudes of the angular movements of the lumbar spine and pelvis could be detected after 4 min of treadmill walking, or of the absolute difference scores beyond 2 min of treadmill walking, in either normal or slow-speed walking groups. CONCLUSIONS: Angular movements of the lumbar spine and pelvis were consistent after 4 min of treadmill walking by normal subjects, both at self-selected and slow walking speeds, indicating that reliable measurements of these parameters could be taken at that time. PMID- 11415665 TI - In vitro patellofemoral joint force determined by a non-invasive technique. AB - OBJECTIVE: To develop a method of measuring the magnitude, direction and point of application of the patellofemoral force (PFF), directly and non-invasively in three dimensions. DESIGN AND METHODS: The compressive PFF is replaced exactly with a tensile force applied to the front of the patella. The magnitude, direction and point of application of the tensile force are then measured. The technique was applied to six normal knees mounted in a 6 degree of freedom rig with quadriceps tendon tension force (QTF) applied to balance a flexing load and to simulate weight bearing. RESULTS: The PFF was greater than in previous more invasive in vitro studies but the results correlated well with recent theoretical analyses. At 20 degrees knee flexion the force was 75% of QTF. It increased to 100% of QTF at 60 degrees knee flexion and remained at this level at higher angles of flexion. The lateral vector of the PFF was small compared to the sagittal plane vector and became negligible beyond 60 degrees of knee flexion. The point of application of the PFF to the patella moved proximally and medially with knee flexion. CONCLUSIONS: A new and reliable method of measuring PFF non invasively and in three dimensions has been developed. RELEVANCE: A new technique is described for measuring the PFF in vitro. The non-invasive nature of the technique makes it useful for studying the effect on the PFF of simulated pathological conditions, surgical procedures and different types of knee replacement. PMID- 11415666 TI - Tibiocalcaneal motion during running, measured with external and bone markers. AB - OBJECTIVE: The purpose of this study was to compare tibiocalcaneal motion during running based on skeletal markers with tibiocalcaneal motion based on external markers. DESIGN. IN VIVO: measurements of external and skeletal tibiocalcaneal kinematics. BACKGROUND: External (shoe, skin) markers are typically used to determine rearfoot kinematics. However, it is not known if such markers are able to provide a good representation of the skeletal (tibiocalcaneal) kinematics. METHODS: Bone pins were inserted into the tibia and calcaneus of five subjects. The 3-D motion of markers attached to bone pins as well as of external markers attached to the shank and shoe were determined during the stance phase of five running trials. Intersegmental motion was expressed in terms of Cardan angles (plantarflexion/dorsiflexion, abduction/adduction, inversion/eversion). RESULTS: It was found that the skeletal inversion/eversion, abduction/adduction, and plantarflexion/dorsiflexion motions were similar across the subjects. The shape of the tibiocalcaneal rotation curves based on external markers were similar to those based on bone markers. However, the rotations were generally overestimated when using external markers, e.g. the average maximal eversion motion calculated from external markers was 16.0 degrees whereas the skeletal maximal eversion motion was only 8.6 degrees. These discrepancies were mainly due to the relative movement between shoe markers and underlying calcaneus. CONCLUSIONS: External markers are only gross indicators of the skeletal tibiocalcaneal motion. The rotations derived from external shoe and shank markers typically overestimate the skeletal tibiocalcaneal kinematics. RELEVANCE: Quantitative results determined from external markers have to be used with caution. For tibiocalcaneal rotations, external markers may be used to show trends, but absolute values cannot be trusted. PMID- 11415667 TI - Measurement of bone lengthening forces; an experimental model in the lamb. AB - OBJECTIVE: To obtain the mechanical behaviour pattern of the lengthening process. DESIGN. IN VIVO: measurement of forces during bone lengthening in lambs. BACKGROUND: A series of parameters of a mechanical and biological nature have a bearing on all lengthening processes, and most of them are not fully understood. METHODS: A strain-gauge-monitored unilateral fixator was designed and used to obtain data about the changes which took place in the forces of elongation at a rate of 1 mm/day in four lambs while a 3 cm progressive lengthening of the left tibia was being performed, analysing how these forces behaved from day to day, and how they changed in the course of a single day. RESULTS: The maximum forces in all the animals each day occur after distraction, and the forces reach their greatest magnitude between days 21 and 25 after surgery, attaining values of slightly over 8 kg (40-50% of the animal's weight). The maximum daily force starts to drop 1 h after distraction, and continues to decrease gradually throughout the day until it reaches a value slightly greater than the initial force on the previous day. CONCLUSION: This pattern is due to the distraction of soft tissues which gradually adapt to their new situation, thereby reducing the level of stress. RELEVANCE: In the daily bone lengthening procedure, the greatest forces are produced in a short period of time immediately after lengthening. they could be reduced to decrease pain in the patient and loads on the device by performing lengthening over a greater number of steps or using dynamic equipment able to absorb these forces. PMID- 11415668 TI - Arm and trunk posture during work in relation to shoulder and neck pain and trapezius activity. AB - OBJECTIVE: To investigate work technique in relation to work-related shoulder and neck pain (SNP) and upper trapezius muscle activity. DESIGN: A matched pair, case control field study of female employees with and without SNP. BACKGROUND: It has proved difficult to distinguish subjects with SNP from those without by vocational electromyographic recordings from the upper trapezius muscle. Other potential risk indicators include psychosocial factors and work technique. This study focuses on the latter. METHODS: Manual (14 pairs) and office workers (24 pairs) were recorded during a 30-min work period. Simultaneous recordings of upper trapezius activity by surface electromyography and arm and upper back postures by inclinometers were analysed. RESULTS: Cases and controls were not differentiated on the basis of arm elevation or of trunk posture in the sagittal plane. No significant correlations were found between variables averaging the muscle activity and the arm elevation over the recording period. Statistically significant correlations were, however, found between these variables when analysing recordings at high time resolution (0.2 s) and adjusting for the delay in arm elevation relative to the upper trapezius muscle activity (r = 0.43, manual group; r = 0.32, office group). CONCLUSIONS: Factors other than arm elevation probably contribute more significantly to the load in the upper trapezius muscle, and to the development of work-related SNP in work situations with moderate arm elevation. RELEVANCE: The study suggests that arm and trunk posture recordings are not a sensitive indicator of risk of shoulder and neck complaints in work with low to moderate biomechanical demands. PMID- 11415669 TI - Roles of deltoid and rotator cuff muscles in shoulder elevation. AB - OBJECTIVE: The objective of this study was to measure abduction moment arms of the supraspinatus, subscapularis, infraspinatus, and deltoid (anterior, middle, and posterior portions) muscles during humeral elevation in the scapular plane (abduction). DESIGN: Moment arms were measured by conducting an in vitro experiment. BACKGROUND: The moment arm of a muscle represents its mechanical advantage, which is an important determinant of muscle function. METHODS: Measurements were made on 10 fresh frozen cadaveric specimens. Tendon excursions were measured as the humerus was elevated in the plane of the scapula. The principle of virtual work was used to estimate the muscle moment arm of each muscle by computing the slope of the tendon excursion versus joint angle relationship. RESULTS: Moment arms were affected by joint angle in a non-linear fashion. The anterior deltoid, middle deltoid, subscapularis, and infraspinatus muscles had abduction moment arms throughout most of the range of motion studied. The posterior deltoid had an adduction moment arm. Internal and external humeral rotation affected the elevation moment arms of all six muscles. CONCLUSIONS: Abduction moment arm magnitudes of the muscles studies vary throughout the arc of elevation. This study was limited by considering broad muscles to have a single line of action. RELEVANCE: The positive elevation moment arms of the infraspinatus and subscapularis muscles indicate that they can elevate the arm in addition to acting as stabilizers. Thus this study suggests a biomechanical explanation for the clinical success of conservative treatment for rotator cuff tears. PMID- 11415670 TI - Forearm extensor and flexor muscle exertion during simulated gripping work -- an electromyographic study. AB - OBJECTIVE: To study forearm muscular exertion during intermittent gripping work at three different regimes resembling vocational work. DESIGN: An electromyographic laboratory study of forearm fatigue during intermittent gripping work at 25% MVC with 10/10, 20/10 and 30/10 s of work/rest was performed. BACKGROUND: Data from the literature indicate that forearm flexors as well as extensors are activated during gripping. However, no systematic quantitative assessment of the muscular exertion has been made hitherto. METHODS: Nine female subjects participated. EMG was detected from five forearm muscles, three extensors and two flexors. Zero crossing rate alterations were used as an index of fatigue. RESULTS: The results show significant (P < 0.05) fatigue signs in at least two of the extensor muscles at all regimes while significant signs of fatigue was seen only at the 30/10 regime for the flexor muscles. CONCLUSIONS: Fatigue effects are generally larger on the extensor side. None of the studied regimes is acceptable from EMG fatigue point of view. RELEVANCE: The muscular fatigue of the forearm extensors in relation to the flexors during gripping work has not been systematically assessed before. Forearm fatigue distribution is probably of great importance for the understanding of forearm disorders related to muscular exertion. PMID- 11415671 TI - Intracarpal canal pressures: the role of finger, hand, wrist and forearm position. AB - OBJECTIVE: The study examined the change in intracarpal canal pressure (ICCP) in relationship to finger, hand, wrist and forearm position. DESIGN: The study was an in vivo measurement of ICCP in seven subjects undergoing a standardized set of manoeuvres that systematically varied finger, hand, wrist, and forearm position. BACKGROUND: It has been known that the ICCP increased with extremes of wrist flexion and extension but the change in pressure in response to radial and ulnar deviation as well as hand and forearm position has not been reported. METHODS: The ICCP was measured using a slit catheter technique; each variation of position was repeated three times with continuous monitoring of ICCP, wrist angulation, and metacarpal-phalangeal joint angulation. RESULTS: The study demonstrated that ICCPs were lowest when the wrist is in a neutral position, the hand relaxed with fingers flexed and the forearm in a semi-pronated position. Wrist extension and flexion resulted in the greatest increase in ICCP followed by forearm pronation and supination. Radial and ulnar deviation also increased the pressure but to a lesser extent. CONCLUSIONS: The findings of this study support the concept that the wrist and forearm should be maintained in a neutral position during vocational and avocational activities in an effort to minimize pressure within the carpal tunnel and thereby reduce the risk of developing carpal-tunnel syndrome. RELEVANCE: It is desirable to know how the ICCP changes in response to change in hand, wrist, and forearm position so that work activities are designed to minimize the pressure within the carpal canal and thus maintain the viability of the median nerve within the carpal canal. PMID- 11415672 TI - Anticipatory postural adjustments during standing in below-the-knee amputees. AB - OBJECTIVE: We studied the role of adaptive changes within the central nervous system in anticipatory postural adjustments seen in unilateral below-the-knee amputees. DESIGN: Changes in electromyographic and mechanical variables were compared during standardized tasks performed by standing subjects. BACKGROUND: Anticipatory postural adjustments represent an important mechanism of postural control which was expected to be changed in amputees because of both mechanical and secondary, neurological reasons. METHODS: Six patients after a below-the-knee amputation and six control subjects stood on a force platform and performed fast bilateral shoulder movements and also dropped or caught a load from (into) extended hands. Anticipatory changes in the background activity of postural muscles were analysed. RESULTS: In amputees, there were cases of marked asymmetry in anticipatory changes of the background muscle activity which were larger on the intact side of the body but were commonly small or absent on the side of the amputation. This asymmetry could lead to larger mediolateral forces and displacements of the centre of pressure. CONCLUSIONS: We suggest that asymmetrical patterns of anticipatory postural adjustments reflect central adaptive changes secondary to the amputation. Rehabilitation approaches would benefit from understanding and taking advantage of the adaptive changes within the central nervous system. RELEVANCE: We demonstrated asymmetries in patterns of anticipatory postural adjustments during voluntary arm movements and load manipulations by standing unilateral amputees. This finding is of potential importance for rehabilitation of amputees and their prosthetic training. PMID- 11415673 TI - Dynamic plantar pressure distribution measurements in hemiparetic patients. AB - OBJECTIVE: A comparison of plantar pressure distribution of hemiparetic patients with a control group was performed to determine quantitative, objective and reproducible criteria for better assessment of hemiparetic gait. DESIGN: This empirical, descriptive study used a clinical sample of 18 hemiparetic patients and compared the data to previously published data from 111 healthy persons. BACKGROUND: Several biomechanical methods have been used in the past to evaluate and classify hemiplegic gait, for example kinetic, cinematographic, or electromyographic systems, but plantar pressure distribution measurement has not been studied. METHODS: Peak pressures were determined under the feet of 18 hemiparetic patients during stance phase using a capacitive pressure distribution platform (EMED-F01 system, Novel GmbH). RESULTS: Hemiparetic patients showed considerably lower peak pressures under all anatomical structures and an unexpected medial load shift in the forefoot. CONCLUSIONS: The grade of spasticity seems to be the most important cause for expression of the medial load shift. Individual pressure distribution gait analysis, as shown in an example, may improve assessment and therapy of hemiparetic patients. RELEVANCE: Plantar pressure distribution data from hemiparetic patients can be used for an analysis of lower extremity dysfunctions. This method may also be employed to assess objectively the success of drug treatment and/or other rehabilitation processes. In particular, individual plantar pressure data from patients will provide additional clinical information for rehabilitation physicians, therapists, and engineers. PMID- 11415674 TI - Preferred placement of the feet during quiet stance: development of a standardized foot placement for balance testing. AB - OBJECTIVE: To establish a standardized stance position for balance testing based on average preferred foot placement, and to compare this to existing standards. DESIGN: Cross-sectional study. BACKGROUND: It has been shown that the orientation of the feet can have a marked influence on the results obtained during balance testing, prompting the need for standardized foot positioning. Unfortunately, current recommendations do not appear to address concerns about the potential effects of 'uncomfortable' or 'unnatural' foot positions on the control of stabilizing reactions. METHODS: The present study identifies the central tendency and variance of the preferred stance width and foot angle, measured from foot tracings in 262 subjects (89 male, 173 female) ranging in age from 19 to 97 years. RESULTS: Results revealed a great degree of variability in preferred stance width and angle across subjects, although mean differences due to gender or age ( 60) were small. The average preferred foot position was 0.17 m between heel centres, with an angle of 14 degrees between the long axes of the feet. Existing standards for stance position lie well outside the range of preferred foot placement. CONCLUSIONS: The wide range of preferred foot placements clearly highlights the need for standardization during balance testing. A standard based on average preference would reduce potential effects of 'uncomfortable' or 'unnatural' foot positions, in comparison to existing standards. RELEVANCE: The development of a standardized stance position for balance testing is necessary since foot placement can influence stabilizing reactions. The present results provide a standardization based on average preferred foot placement, which will minimize between-subject variability and reduce the abnormal constraints that are placed on subjects by existing standards. PMID- 11415675 TI - The torsional strength of bones with residual screw holes from plates with unicortical and bicortical purchase. AB - OBJECTIVE: To evaluate the effect of unicortical and bicortical screw holes on residual bone strength by comparing the in vitro torsional strength of cadaveric sheep tibiae with screw holes from plates with unicortical and bicortical purchase relative to each other and to intact bone. DESIGN: The paired tibiae were grouped randomly and torsion tested to failure as follows: Group I -- unicortical screw holes versus intact bone; Group II -- bicortical screw holes versus intact bone; and Group III -- bicortical versus unicortical screw holes. BACKGROUND: Recently the point contact fixator, or PC-Fix (using screws with unicortical purchase), was designed to minimize bone devascularization beneath the plate compared to the conventional dynamic compression plate, or DCP (bicortical purchase), and possibly reduce refracture rates after plate removal. However, the effects of unicortical versus bicortical residual screw holes on potential refracture are unknown. METHODS: Screw holes were drilled and tapped through either a 7-hole bicortical DCP or a unicortical PC-Fix in 18 paired cadaveric sheep tibiae. The bones were then tested in torsion. RESULTS: The average decreases in the torsional strength of unicortical screw holes versus intact bone, bicortical screw holes versus intact bone, and bicortical versus unicortical screw holes were 21.6, 31.4, and 26.7% respectively. Mean torque values to failure were statistically significant within each of the three groups: P<0.01, 0.001, and 0.001 respectively. CONCLUSIONS: Bones with unicortical screw holes are significantly weaker in torsion than intact bones, but significantly stronger than bones with bicortical holes. RELEVANCE: Refracture after hardware removal is a recognized clinical complication of the traditional bicortical screw fixation. In combination with the demonstrated biological advantages, implants with unicortical purchase may be associated with less risk for early refracture upon removal. PMID- 11415676 TI - Three-dimensional primary stability of cementless femoral stems. AB - OBJECTIVE: This study investigates by means of a new bone-prosthesis interface motion detector whether conceptual design differences of femoral stems are reflected in their primary stability pattern. DESIGN: An in vitro experiment using a biaxial materials testing machine in combination with three-dimensional motion measurement devices was performed. BACKGROUND: Primary stability of uncemented total hip replacements is considered to be a prerequisite for the quality of bony ongrowth to the femoral stem. Dynamic motion as a response to loading as well as total motion of the prosthesis have to be considered under quasi-physiological cyclic loading conditions. METHODS: Seven paired fresh cadaveric femora were used for the testing of two types of uncemented femoral stems with different anchoring concepts: CLS stem (Spotorno) and Cone Prosthesis (Wagner). Under sinusoidal cyclic loading mimicking in vivo hip joint forces a new measurement technique was applied allowing for the analysis of the three dimensional interface motion. RESULTS: Considerable differences between the two prostheses could be detected both in their dynamic motion and total motion behaviour. Whereas the CLS stem, due to the wedge-shaped concept, provides smaller total motions, the longitudinal ribs of the Cone prostheses result in a substantially smaller dynamic motion. CONCLUSIONS: The measuring technique provided reliable and accurate data illustrating the three-dimensional interface motion of uncemented femoral stems. PMID- 11415677 TI - A biomechanical study on five unilateral external fracture fixation devices. AB - OBJECTIVE: This study examines plastic, slip and fatigue failures in external fixator frames. DESIGN: Failure was examined in five commonly used unilateral external fixators using experimental models that simulated both stable and unstable diaphyseal fractures. BACKGROUND: Plastic failure arises from permanent deformation of a frame initiated by bending of frame components, while slip failure arises from slip at clamping interfaces. Such failure may lead to refracture or to malalignment of the bone (transverse or angular). Fatigue failure arises from the loosening or wear of components under long-term cyclic loading; this can lead to variable interfragmentary displacement and impairment of fracture stability. METHODS: Axial loads were measured at which plastic or slip failure occurred, as were changes in interfragmentary motion due to 10 000 load cycles simulating walking activity. RESULTS: In four fixators, plastic or slip failure initiated bone malalignment at only 50% of average adult weight bearing (650 N). Additionally, in three fixators a third screw at the clamp centre was found to reduce retention of the two adjacent screws, causing premature slip failure. Evidence of fatigue was found in all the fixators after only 10 000 load cycles. In one fixator, the amplitude of elastic interfragmentary motion increased progressively by 30% transversely, 15% angularly and 100% torsionally. In another fixator it increased by 10% in all directions while the column was locked (non-dynamizing), and it increased a further 10% while it was unlocked and able to slide telescopically (dynamizing). Clamp slip occurred at the column of a third fixator almost immediately after commencing the cyclic loading. CONCLUSION: Plastic or slip failure of frames may occur prematurely during routine weight-bearing on unstable fractures, and frame fatigue may affect long-term interfragmentary stability. PMID- 11415678 TI - Pattern classification reveals intersubject group differences in lumbar muscle recruitment during static loading. AB - OBJECTIVE: This paper examines interindividual differences in the patterns of torso muscle recruitment during 3-dimensional static moment loading of the lumbar spine. DESIGN: A mathematical model (artificial neural network) was used to differentiate individual patterns of muscle response. BACKGROUND: Traditionally, experimental myoelectric data is averaged over subjects, assuming an ideal mean response to a given loading. However, averaging may overlook important information and implications associated with interindividual variability. METHODS: In this study a simple classification tool in the form of a competitive neural network model is developed and used to evaluate lumbar muscle recruitment patterns. RESULTS: Subjects formed consistent and denumerable clusters, and could be categorized as either 'majority' or 'minority' type responders, based on their individual muscle response patterns as discerned from the output of the competitive network model. The practical significance of these differences is shown by comparison of muscle activity with more established optimization-based force predictions. Those subjects categorized as majority-type responders had muscle activity in better correspondence with optimization-based predicted forces. Subjects in minority categories displayed more variance in their response patterns and larger degrees of antagonistic cocontraction. CONCLUSIONS: The implications for deterministic (e.g. optimization-based) biomechanical modelling are discussed. It is speculated that interindividual muscle recruitment differences may be important for assessing individual musculoskeletal risk. PMID- 11415679 TI - Biomechanical assessment of lifting dynamics, muscle activity and spinal loads while using three different styles of lifting belt. AB - OBJECTIVE: To demonstrate the influence of different types of lifting belts on trunk motion, muscle activity and spine loading during symmetric and asymmetric lifting exertions. DESIGN. IN VIVO: measurements were achieved representing lifting dynamics, applied trunk moments and myoelectric activity. Dynamic spinal loads were determined from a validated biomechanical model of lifting. BACKGROUND: There is a great deal of controversy as to whether lifting belts are a benefit or a liability to manual materials-handling activities. A review of the literature demonstrates that there is a large amount of conflicting evidence and few definitive, well-executed studies upon which to base an opinion regarding these devices. METHODS: Fifteen subjects lifted boxes of 14 kg and 23 kg from sagitally symmetric and asymmetric origins to an upright posture. Dynamic trunk motions, lifting moments, myoelectric activity and modelled spinal loads were examined as a function of three belt styles (elastic, leather, and orthotic) and compared with results from a no-belt condition. RESULTS: Lifting belts reduced peak trunk angles, velocities and accelerations in the sagittal, lateral and transverse planes. However, only the elastic belt successfully reduced trunk motions in all three dimensions. The orthotic belt significantly increased the lifting moment associated with a given weight. A minor redistribution in muscle activity was observed when wearing an elastic belt. A statistically significant reduction in spinal load was associated with the elastic belt. However, a great deal of variability between subjects was noted. Some subjects experienced increased spinal load while wearing the elastic lifting belt. CONCLUSIONS: These results demonstrate that the biomechanical operation of lifting can be influenced by the type of lifting belts used. PMID- 11415680 TI - On the mechanical effects of knee bandages in the therapy of patellar chondropathy. AB - OBJECTIVE: The study aimed to clarify whether the therapeutic success of an infrapatellar bandage placed above the tibial tubercle in the treatment of patellar chondropathy is based on altered knee joint loads during specific movements. DESIGN: In a clinical trial the influence of the bandage on kinetic and EMG variables is investigated. BACKGROUND: Although the used bandage is not a supporting device, the patients report an instant improvement. Thus neurological mechanisms are hypothesized as capable of pain reduction by changing the neuromuscular movement coordination or sensation thresholds. METHODS: Ten patients performed three different movements (running, drop jump, walking downstairs), before and after attaching the bandage, while kinematic, dynamic, and EMG data were acquired. After calculating the intersegment moment of the knee joint, different kinetic and EMG parameters were combined to the dependent MANOVA procedure. RESULTS: The neuromuscular activity during drop jump was reduced significantly (P < 0.01) wearing the bandage while, apart from individual changes, no general alteration of the knee joint loads could be statistically confirmed. CONCLUSIONS: The effect of the examined infrapatellar bandage is not a consequence of a decreased joint load due to a general movement adaptation, but is probably due to a neural influence on the nociception. PMID- 11415681 TI - Dynamic testing of the motor stereotype in prone hip extension from neutral position. AB - OBJECTIVE: The purpose of the study was to identify a sequential activation of lumbar and hip muscles in active prone hip extension from neutral position in subjects without a history of low back pain. DESIGN: Using surface electromyography, the myoelectric activity onsets of agonistic and antagonistic hip muscles were recorded. BACKGROUND: The development of low back pain is ascribed to changes of the muscle firing order in prone hip extension. There appeared to be no normative data on muscle firing order of the lumbar and hip musculature to provide a basis for recognizing variations. METHODS: Fifteen healthy subjects performed standardized right hip extensions. Time-normalized onset of EMG activity in each muscle was measured for each trial. RESULTS: A MANOVA revealed significant (P < 0.01) differences between the activity onsets of synergistic muscles in hip extension. CONCLUSIONS: On the results it was concluded that the consistent muscle firing order of ipsilateral lumbar erector spinae, semitendinosus, contralateral lumbar erector spinae, tensor fasciae latae, and gluteus maximus demonstrate the characteristic pattern in prone hip extension. In future studies it is essential to evaluate also the hip extension pattern in functional upright conditions during gait and their relationship to spinal mechanics. PMID- 11415682 TI - A three-dimensional multivariate model of passive human joint torques and articular boundaries. AB - OBJECTIVE: The development of a novel three-dimensional mathematical model of passive human joint torques that is of practical use, takes into account the complex non-linear interactions that exist between the moments generated by the various passive structures spanning the joint in question, and is applicable to all types of articular joints. DESIGN: Mathematical model validated by practical implementation. BACKGROUND: Previous models of passive human joint torques were predominantly one-dimensional and did not take into account the interactions of the various structural components. METHODS: Mathematical modelling is used in conjunction with repeated passive torque experiments on the right elbow joint of a healthy 29-year-old male. On the basis of the experimentally observed torque angle data, the full set of articular model parameters characterizing the two dimensional passive elastic torque function of the subject's right elbow joint could be determined. RESULTS: In addition to the passive elastic torque functions, the two-dimensional contour of the elbow joint's articular boundaries was also obtained. CONCLUSIONS: The present model is especially useful for assessing certain pathological conditions in any body joint investigated, and in addition is best suited for the inclusion in large-scale dynamic simulation models of the human neuromusculoskeletal system. PMID- 11415683 TI - Effects of contract-relax stretching procedures on active range of motion of the cervical spine in the transverse plane. AB - OBJECTIVE: To determine if a short regime of stretching exercises could affect cervical range of motion (RoM). DESIGN: A single-blind trial design was adopted. BACKGROUND: Stretching has become important with respect to sports as well as health care. Although much is surmised about the effects of stretching exercise on spinal RoM, little has been published, those that have using symptomatic subjects. Therefore it is an appropriate time to study this subject more closely. METHODS: Forty volunteer, asymptomatic male subjects were assigned into control (n = 19) or exercise (n = 21) groups and asked not to vary their daily routines too much. The exercise group performed stretching exercises twice a day, on days 1-7 inclusive. All subjects performed warm-up exercises (active maximal rotation and lateral flexion in the cervical spine) before assessing active cervical RoM on days 1, 3, 7 and 14, using a cervi-corotometer, whose least significant difference for left, right and total cervical rotation were 3.8 degrees, 3 degrees and 5.2 degrees ). RESULTS: The exercise group had a significantly increased active cervical RoM by day 7 (c. 14%; 22 degrees for total rotation). This change reverted towards pre-stretch values 7 days after exercises had been discontinued. CONCLUSIONS: Performing stretching exercises increases cervical RoM in the short term. These effects wear off rapidly if the stretching regime is discontinued. PMID- 11415684 TI - Body height changes with hyperextension. AB - The purpose of this study was to determine if the overall body height, as measured by a stadiometer, could be increased by brief episodes of hyperextension rather like a stretch that people frequently employ when arising. The subjects were loaded with 10 kg and the recovery with quiet sitting was compared to hyperextension 'stretches'. It was found that 15 s of hyperextension caused a significant temporary height increase. PMID- 11415685 TI - Finite element analysis of human knee joint in varus-valgus. AB - OBJECTIVE: The overall response, load transmission, role of ligaments, and state of stress in various components under varus-valgus moments in the intact and collateral-deficient tibiofemoral joint are investigated. DESIGN: A non-linear finite element model consisting of bony structures (tibia and femur), their articular cartilage layers, medial and lateral menisci and four primary ligaments (cruciates and collaterals) is utilized. BACKGROUND: Valgus and varus stresses are among the primary mechanisms of injury to knee ligaments. Several in vitro studies have investigated the role of ligaments in resisting such loads and on the way deficiency in either collateral may affect the response. METHODS: Cartilage layers are isotropic while menisci are non-homogeneous composite. The articulation of cartilage layers with each other and with the intervening menisci and the wrapping of the medial collateral ligament around the tibial edge are treated as large displacement frictionless contact problems. The non-linear elastostatic response of the joint at full extension is computed under varus valgus moments applied to the femur with the tibia fixed. Cases simulating deficiency in collaterals and constraint on femoral axial rotation are also studied. RESULTS: The response is non-linear with large coupled axial rotations, internal in varus and external in valgus. In intact and collateral-deficient states, the joint shows varus or valgus openings so that the articulation occurs at one plateau only, medial in varus and lateral in valgus. Large tensile forces in cruciates in collateral-cut models generate higher compression penalty on the loaded plateau. CONCLUSIONS: Collaterals are the primary load-bearing structures; their absence would substantially increase primary laxities, coupled axial rotations, forces in cruciates, and articular contact forces. Good agreement with measurements is found. RELEVANCE: Detailed knowledge of joint biomechanics is essential in the diagnosis, prevention and treatment of observed disorders. Absence of collateral ligaments increases the loads in cruciates and contact stresses transmitted through cartilage layers and menisci, and thus places the affected components at more risk, especially when varus-valgus is accompanied by other modes of loading as well. PMID- 11415686 TI - Bioabsorbable pin fixation of intercarpal joints: an evaluation of fixation stiffness. AB - OBJECTIVE: Bioabsorbable pins made of poly-p-dioxanone (Johnson and Johnson Orthopaedics, Raynham, Massachusetts) were compared to steel pins in the fixation of the scapholunate and lunotriquetral intercarpal joints of the wrist. DESIGN: An in vitro experiment was performed using fresh-frozen cadaver wrists. BACKGROUND: In the surgical treatment of wrist ligament injuries, temporary intercarpal joint fixation is required. Although steel pins are currently used, there are several clinical problems associated with their use. These complications could potentially be reduced by using bioabsorbable fixation. METHODS: The carpal bones were mounted between sets of plates which were then attached to a materials testing machine. Two parallel pins were used for joint fixation; steel pin fixation was tested first in each joint. Seven scapholunate joints and seven lunotriquetral joints were each tested in three different modes: translation in two orthogonal directions, and rotation. RESULTS: Bioabsorbable pin fixation provided an average of 46% of the stiffness of steel pin fixation. CONCLUSIONS: Although bioabsorbable pin fixation was statistically less stiff than steel pin fixation, relatively high loads were required to induce significant intercarpal joint translation and rotation. RELEVANCE: The strength retention profile of poly-p-dioxanone pins and the quality of fixation demonstrated in this study indicate that these bioabsorbable pins could provide satisfactory fixation of the scapholunate and lunotriquetral joints in the treatment of partial injuries of the intercarpal ligaments. PMID- 11415687 TI - Load dependence in carpal kinematics during wrist flexion in vivo. AB - OBJECTIVE: The hypothesis tested was that generation of torque at the wrist affects joint kinematics. DESIGN: An in vivo study of normal wrist kinematics during plantar flexion motion against a constant load was undertaken, using a custom-designed instrumented apparatus to track the motion of the hand during the task. BACKGROUND: Despite clinical observations of a relationship between motion loading and pain in wrists affected by rheumatoid arthritis, there is little published literature on thein vivo kinematics of the normal human wrist under load. METHODS: Ten volunteers with no wrist pathology were tested while generating torques of zero, 1.1 and 2.2 N m in a planar, unidirectional flexion motion. Hand kinematics were computed using the Planar Rigid Body Method algorithm and an 8 degrees angular step size. The finite radius of motion and the range and standard deviation of the residuals to a fitted second-order curve were used as indices of changes in the kinematics. RESULTS: The magnitude of both the range and standard deviation of the residuals were found to increase significantly with torque at the 95% confidence level. CONCLUSIONS: The wrist does not behave like a smooth mechanism when generating torque. Load affects carpal kinematics. RELEVANCE: We propose that fluctuations in the finite radius of motion are the natural kinematic consequence of intercarpal motion known to occur during wrist flexion. Wrist kinematics may be particularly sensitive to load and joint integrity because orchestrated intercarpal motion depends on the soundness of articular and ligamentous structures, the first to be affected in joint degenerating conditions such as rheumatoid arthritis. Thus, wrist kinematics under load may be a key to characterizing joint integrity. In wrist pathologies, simple planar testing of carpal kinematics under reproducible and controlled joint torque conditions may be a useful way to assess joint involvement before the onset of gross dysfunction, and to evaluate treatment outcomes. PMID- 11415688 TI - Biomechanical study of stress in the fifth metatarsal. AB - OBJECTIVE: The stress throughout the fifth metatarsal was determined under various loading conditions, in order to better understand the causes of fractures to this bone. DESIGN: A mathematical approach was taken, in which the stresses were analysed using the methods of beam theory. BACKGROUND: Finite element analysis has frequently been used to determine the stress in bones. Beam theory provides an easier method for determining the force and moment resultant in any cross-section. The distribution of stress throughout the cross-section can then be found by solving certain partial differential equations defined on the cross sections. METHODS: Cross-sections of the bone were obtained by slicing a mould, into which the bone was placed, at numerous intervals along its length. Analytic expressions describing each cross-section were obtained by fitting a Fourier series to a sequence of points along the boundary. RESULTS: The maximum stress found in the fifth metatarsal resulted from an oblique load, and had a magnitude less than would occur in a subject during normal walking. CONCLUSIONS: Since the magnitude of the stress is submaximal, this study lends theoretical support to the clinical observation that the diaphyseal fracture is indeed a stress fracture. RELEVANCE: Our analysis adds a biomechanical rationale to the pathomechanics of diaphyseal stress fractures of the fifth metatarsal. It suggests that inversion during repetitive activities may predispose the foot to fractures at a predictable location. PMID- 11415689 TI - Surface electromyography in the identification of chronic low back pain patients: the development of the flexion relaxation ratio. AB - OBJECTIVE: To develop a reliable and repeatable way to monitor changes in the flexion relaxation phenomenon of the lumbar paraspinal muscles during forward flexion by the development of a flexion relaxation ratio and observation of the sEMG activity in standing and during forward flexion in patients with chronic low back pain (CLBP) and healthy controls. DESIGN: Two experiments were conducted, the first to assess the test-retest reliability of the measure in a group of CLBP (n = 11) patients; the second compared the results between a group of normal healthy controls (n = 20) and a group of CLBP patients (n = 70). RESULTS: Repeated measurements over 4 weeks demonstrated between session reliability of between 0.81 and 0.98 for the dynamic activity. The levels of sEMG activity in the fully flexed position was significantly greater in the fully flexed position in the CLBP group than the controls. The flexion relaxation ratio (FRR), a comparison of the maximal sEMG activity during 1 s of forward flexion with activity in full flexion, demonstrated significantly lower values in the CLBP than the control group. The combined discriminant validity for the FRR for all four sites resulted in 93% sensitivity and 75% specificity. CONCLUSION: The FRR clearly discriminated the patients from the healthy controls. These results indicate that dynamic sEMG activity of the paraspinal muscles can be reliably measured and is useful in differentiating CLBP patients from normal controls. RELEVANCE: Analysis of the pattern of different levels of muscle activity during a forward flexion can be used in CLBP where normalization of the sEMG signal to the maximum voluntary contraction may be difficult. The FRR may be used in the assessment of change in the flexion relaxation phenomenon following treatment interventions. PMID- 11415690 TI - Selective activation of the external oblique musculature during axial torque production. AB - OBJECTIVE: To investigate whether different geographical regions of the external oblique musculature can activate at different levels and, if they do, to quantify the magnitude of these differences as a function of postural parameters during twisting exertions. DESIGN: Repeated measures design using electromyography on healthy subjects. BACKGROUND: The majority of the models currently used to assess spinal loading have represented the trunk musculature using single force vectors connecting a muscle's point of origin to its point of insertion. However, for muscles with large areas of origin and/or insertion (such as the external obliques), this single vector modelling approach misrepresents the multiple vector reality which, in turn, underestimates the complex loads these muscles can develop. METHODS: Nine subjects performed sub-maximal isometric axial twisting exertions (20, 40 and 60% of maximum voluntary contraction) while assuming six different postures defined by three levels of axial rotation (-20 degrees, 0 degrees and 20 degrees ) and two levels of sagittal flexion (0 degrees and 20 degrees ). As the subjects performed these isometric exertions, the integrated electromyographic activity was sampled using surface electrodes at five different locations over the right and the left external oblique muscles. RESULTS: The results showed significant (p<0.05) regional differences in the activation profiles and these activation profiles changed as a function of trunk posture. CONCLUSIONS: The external oblique musculature is capable of differential activation and the activation profiles of the different regions are affected by the posture of the torso. RELEVANCE: These findings suggest that the external oblique muscle is capable of selective activation of different regions along its cross-section and should, therefore, be modelled using multiple vectors. The result can have a direct bearing on the calculated spine loading, especially lateral and anterior/posterior shear forces. PMID- 11415691 TI - Training does not increase maximal lumbar extension in healthy adolescents. AB - OBJECTIVE: To investigate if there is training reserve in the maximal lumbar extension. DESIGN: Three-year longitudinal study. BACKGROUND: Among adults there is variation in the normal range of sagittal motion of the lumbar spine, but reduced spinal flexibility does not predict future occupational back pain. In various sports and in ballet, maximal extension of lumbar spine is a common manoeuvre, and low-back pain is also common. It is not known whether training increases maximal extension of healthy back. Forceful training of maximal extension may injure the anatomical structures limiting the extension range. METHODS: We compared lumbar sagittal flexibility to hip flexor and hamstring flexibility in a 3-year longitudinal study on female ballet dancers (n = 18), athletes (n = 31), and controls (n = 17) before and after their adolescent growth spurt. RESULTS: Ballet dancers had more flexible hamstrings and hip flexors than controls, but there were no group differences regarding the maximal lumbar flexion or extension. These results persisted throughout follow-up. CONCLUSIONS: The maximal physiological extension of the lumbar spine cannot be increased by training in healthy adolescents. RELEVANCE: An attempt to exceed the physiological maximum extension may only cause overly hard strain on specific anatomical structures of the lumbar spine. This knowledge should be considered when the rules of sports and choreography of dance performances are considered. PMID- 11415692 TI - A non-invasive protocol for the determination of lumbosacral vertebral angle. AB - OBJECTIVE: The purpose of this study was to develop a non-invasive method to predict the lumbosacral and vertebral angles. DESIGN: The candidate predicting variables were the externally measured marker angle, pelvic angle, and lumbosacral angle. BACKGROUND: Several investigators used surface rigid markers to represent the angles of lumbosacral vertebrae. Previous studies show a high level of validity for measuring the lumbar curvature during stance and for obtaining the vertebral angle in the lumbar region. However, our review of literature failed to find a data transformation model illustrating the lumbosacral vertebral angle using external measurements as predictors. METHODS: Sixteen healthy male subjects participated in the radiographic and videographic measurements. Data of 12 of the 16 subjects were used for model development, and data of the other four subjects were used for model validation. RESULTS: The results showed significant differences between the directly measured vertebral angle from the X-ray film and the externally measured angles of the surface markers at the L(5) and S(1) levels (P<0.05). Linear regression models for calculating vertebral angles were developed with R(2) values of 0.97, 0.98, 0.91, and 0.92 for the vertebral levels of L(1), L(3), L(5), and S(1) respectively. The validation result showed that there was no significant difference between the calculated and the X-ray data. CONCLUSIONS: The protocol of using the skin surface rigid markers and the predicting models was justified in the study to provide a simple and valid non-invasive method for the recording of the internal vertebral angle in the sagittal plane. RELEVANCE: Measurements of lumbosacral vertebral angle are important in assessing the stresses acting on the low back in lifting. The results of this study provided a simple and valid non-invasive method by rigid markers for the prediction of internal vertebral angles under various trunk positions. PMID- 11415693 TI - Methodological considerations for using inductive sensors (3SPACE ISOTRAK) to monitor 3-D orthopaedic joint motion. AB - OBJECTIVE: To assess and improve methodological constraints of the 3SPACE ISOTRAK. DESIGN: Several data sets were used to evaluate the following constraints: the performance is adversely affected by close proximity of metallic objects or electromagnetic fields; the3SPACE field contaminates electromyography recordings; and cross-talk from one axis to the other two occurs if the Euler rotation sequence is not matched to the orthopaedic description and function of the joint in question. BACKGROUND: The 3SPACE ISOTRAK is one of the few instruments suitable for measurement of relative three-dimensional joint rotation, however several methodological constraints exist. RESULTS AND CONCLUSIONS: Generally, accuracy can be maintained by restricting offending objects from the sensitive zone between the transmitter and sensor and using long leads to afford the subject distance from computers and other electrical instrumentation; while contamination of EMG amplitude in the time domain can be minimized by using quality instrumentation, and on occasion modifying electrode orientation. Contamination of the EMG frequency content appears unavoidable at low levels of muscle contraction; and cross-talk between axes is minimized by matching the first Euler rotation with the axis of the joint that is characterized by the greatest expected angular displacement. RELEVANCE: The 3SPACE ISOTRAK is one of the few instruments available that is suitable for measuring three axes of relative joint motion (local joint axes that rotate with the joint -- not global). This paper suggests several techniques to increase the accuracy and viability of this instrument. PMID- 11415694 TI - The Contribution of Biomechanics to the Prevention and Treatment of Low Back Pain: 1st International Workshop. PMID- 11415695 TI - In vitro and in vivo radiomorphometric analyses of distal screw hole position of the solid tibial nail following insertion. AB - OBJECTIVE: The objective of this study was to analyse the position of the distal interlocking holes of an unreamed solid tibial nail relative to the adjacent cortices of the tibial bone. DESIGN. IN VITRO: and in vivo radiographic morphometric analysis. BACKGROUND: To date, 'radiation-independent' intramedullary distal interlocking aiming systems have not been successful, primarily because proximally mounted aiming devices cannot account for the deformation of the nail during insertion; therefore the distal interlocking holes relative to the adjacent cortices must be understood. METHODS: A two-dimensional radiographic-morphometric analysis was performed in 40 tibiae after in vitro (n = 20) and in vivo (n = 20) implantation of unslotted stainless steel unreamed 8- and 9-mm diameter intramedullary nails. RESULTS: The combined in vitro and in vivo results showed average distances between the upper distal transverse hole and the anterior aspect of the tibia of 12.3 mm (SD 1.7 mm; variance 2.9 mm(2); range 7.6 mm). CONCLUSIONS: The distances between the upper distal transverse hole and the anterior aspect of the tibia are the most reliable parameters of distal nail hole position. RELEVANCE: For a 'radiation-independent' distal interlocking aiming system for unreamed intramedullary tibial nails to be successful, it could take advantage of the relatively constant distance between the anterior cortex and the most proximal distal screw hole. PMID- 11415696 TI - NOVEL Award 1996: 2nd prize Tenodeses do not fully restore ankle joint loading characteristics: a biomechanical in vitro investigation in the hind foot. AB - OBJECTIVE: In order to understand the biomechanical consequences of ligament injuries and surgical reconstruction procedures, their effects on intra-articular loading in the ankle joint complex and Chopart joint line and on the plantar pressure patterns were investigated in vitro. METHODS: Twelve fresh-frozen lower leg specimens were freed of soft tissue down to the malleoli and prepared for accessing the talocrural, subtalar, talonavicular and calcaneocuboid joints. The specimens were fixed in a loading simulator and axially loaded with 600 N in six experimental conditions: intact; after cutting the anterior talofibular ligament; after additionally cutting the calcaneofibular ligament; after performing three common types of tenodeses, the Evans, Watson-Jones and Chrisman-Snook procedures. The intra-articular loading characteristics were determined with pressure sensitive film. Plantar loading patterns were measured with a capacitive EMED pressure distribution platform. RESULTS: Average intra-articular pressures were increased and were related either to decreased contact areas or to increased contact forces found in all joints after ligament resections and tenodeses. Plantar loading was increased under the medial aspect of the foot and decreased under the midfoot region. CONCLUSIONS: The results indicate that ankle ligament injuries, as well as surgical reconstructions by tenodeses, affect joint loading characteristics and may exacerbate joint degeneration. RELEVANCE: Excessive laxity of the ankle joint is considered a pre-arthrotic condition and is treated with various surgical procedures. Some of these procedures that utilize the tendon of the peroneus brevis have been shown to change joint kinematics. The aim was to evaluate joint loading characteristics and the potential danger of developing arthritis as a consequence of various tenodeses techniques. For this purpose, in vitro investigations are needed to directly determine intra-articular pressure measurements. PMID- 11415697 TI - Effect of six types of footwear on peak plantar pressures in patients with diabetes and transmetatarsal amputation. AB - INTRODUCTION:: Patients with diabetes (DM) and transmetatarsal amputation (TMA) are at high risk for skin breakdown from excessive peak plantar pressures (PPP). The primary purpose of this study was to determine how footwear (full length shoe or short shoe), a total contact insert, a rigid-rocker bottom (RRB) sole, and an ankle-foot-orthosis (AFO) affect PPP on the distal residuum and contralateral extremity of patients with DM and TMA. A secondary purpose was to monitor various functional measures during use of the footwear. METHODS:: Thirty patients with DM and TMA participated (mean age 62+/-4 years). The mean duration of DM was 19.9+/ 10.1 years, and the mean time since TMA was 27.4+/-28.1 months. The following footwear was provided after a check-out from an orthotist and physical therapist (PT); 1) Full length shoe (ie shoe length prior to surgery), with a toe filler, 2) full length shoe, total contact insert, and an AFO, 2) full length shoe, total contact insert, and an AFO, 3) full length shoe, total contact insert, and a RRB sole, 4) full length shoe, total contact insert, RRB sole, and an AFO, 5) short shoe (ie length of residuum), total contact insert, and RRB, 6) short shoe, total contact insert, AFO, and RRB sole. In-shoe PPP during walking at the distal residuum and forefoot of the contralateral extremity were measured using the F Scan System with established reliability under similar conditions (Generilizability coefficient =.75). Each measurement occurred after a one month adjustment period. Data were analyzed using a univariate repeated measuresANOVA. Individual contrasts were used for post-hoc analysis on those variables showing a significant overall F value (p<.05). RESULTS:: Compared to a regular shoe with a toe-filler, all conditions except the short shoe (#5), resulted in lower PPP on the distal residuum (p<.05). Condition 3, the full length shoe, total contact insert, and RRB resulted in lower pressures on the distal residuum and forefoot of the contralateral extremity compared to a regular shoe and toe-filler, and had few functional complaints as identified by the patient, orthotist or PT (3/27). Footwear using an AFO (Conditions 2,4,6) showed reduced PPP on the residuum, but most patients (16/29) had functional complaints. The short shoe (condition 5) had the fewest[Table: see text] functional complaints (2/26), but did not significantly reduce PPP and had the highest cosmetic refusal rate (5/26). DISCUSSION AND CONCLUSIONS:: Although there are individual patient characteristics which warrant other prescriptions, based on the results of this study, we recommend the full length shoe, total contact insert, and RRB sole for most patients with DM and TMA to reduce PPP. A reduction in PPP should help to lower the high risk of skin breakdown in this patient population. PMID- 11415698 TI - The average pressure distribution of the diabetic foot: can it be used as a clinical diagnostic aid? AB - INTRODUCTION:: The aim of this study was to investigate whether averaged data obtained from pressure distribution measurements could be used to distinguish differences between diabetic feet with normal and no sensation. The calculated parameters most sensitive to any differences were established, and the potential of the averaged maximum pressure picture as a quick recognition pattern of a neuropathic diabetic foot was assessed for clinical use. METHODS:: Dynamic pressure distribution data was collected from 56 diabetics using an emed SF2 platform. The first step method was used, and only one step per subject was recorded for this set of data. Each subject completed a questionnaire which included a grading of the sensitivity to vibration on the plantar surface of the foot evaluated by a clinician. The subjects were subsequently divided into four groups depending on this sensitivity, but only data from the two groups with normal (n=16) and no sensation (n=12) were evaluated in this study. Control data were obtained from two separate sources: for adults (n=40) using an emed SF4 platform and for teenagers (n=73) using a mini-emed platform. The groupmask evaluation program from the novel-win software and the average program from the novel-ortho software were used to evaluate the data. Each dynamic pressure distribution was automatically masked using the foot recognition algorithms of the 'novel' mask, which divided the foot into ten areas: the heel, midfoot, first, second, third, fourth and fifth metatarsal heads (MTH), hallux, second toe, and remaining toes. The parameters analysed for each of these masks included: the mean peak pressure, maximum force, contact time, beginning of contact time, pressure-time integral and force-time integral. RESULTS AND DISCUSSION:: The results showed clearly that both pressure and contact time are important parameters in differentiating a neuropathic from a non-neuropathic diabetic foot. Values of the maximum force in each area of the foot showed no difference between the two groups. Owing to the small sample sizes for the two diabetic groups the 95% confidence limits were relatively large, however, clear differences existed between the two sets of data for the mean peak pressure and the mean pressure-time integral for the total foot and the forefoot. Based upon this data, peak pressure values above 56 N/cm(2) for the 1st MTH, 46 N/cm(2) for the 3rd MTH, 35 N/cm(2) for the 4th MTH and 26 N/cm(2) for the 5th MTH, and pressure-time integrals above 31 Ns/cm(2) for the 1st MTH, 21 Ns/cm(2) for the 2nd MTH, 24 Ns/cm(2) for the 3rd MTH, 19 Ns/cm(2) for the 4th MTH and 12 Ns/cm(2) for the 5th MTH would suggest the possibility of a neuropathic diabetic foot. The averaged maximum pressure distribution picture for each group showed some differences between diabetic feet with normal sensation and those with no sensation. However, owing to the small size of the groups, it was felt that the averaged pressure pictures were, at the moment, limited in their use as a quick recognition pattern for the clinician. Although further data has already been collected and will be added to the databank in the near future. CONCLUSIONS:: These data show that averaged maximum pressure pictures alone do not provide sufficient information to differentiate clearly between a non-neuropathic and a neuropathic diabetic foot. Parameters obtained from the dynamic process, in particular, the pressure-time integral, must also be taken into account. It is anticipated that this work will be continued in order to investigate differences between other dynamic processes of potential importance, such as the location and speed of the gait line. ACKNOWLEDGEMENTS:: The authors would like to thank Dr M Morlock and Dr T Tsvetkova for the control data. PMID- 11415699 TI - The effect of foot orthotics on the initiation of plantar surface loading. AB - INTRODUCTION:: Little information exits concerning the effect, if any, foot orthoses have upon how the foot is loaded during normal walking. The purpose of this study was to determine if there was a difference in the initiation of forefoot loading with different densities of foot orthoses. METHODS:: Ten volunteers (5 men, 5 women) between the age of 23 and 43 years (mean=30.3 yrs) participated in this study. None of the subjects had a history of congenital deformity, hallux valgus, malleolar torsion or a history of injury or pain to either lower extremity 12 months prior to the start of the investigation. Two different pair of foot orthoses were provided to each subject in the study. One pair consisted of UCO-BKS medium width foot orthoses with a durometer rating of 55-60 (Shore A gauge). Forefoot and/or rearfoot wedging was applied to these orthoses if indicated from a physical examination. This orthotic condition was labeled RIGID. The second pair consisted of UCO-BF foot orthoses with a durometer rating of 30-35 (Shore A gauge). This orthotic condition was labeled SOFT. In addition to the foot orthoses, each of the 10 subjects were fitted with the same type of footwear (Reebok Boston Road). Following a 'break-in' period for the footwear and orthotics, each subject walked along a 12 m walkway with the EMED PEDAR insole placed inside their shoes. A total of 10 consecutive walking trials were collected at each subject's self-selected speed. Using the NOVELWin MASKS program, the plantar surface of the forefoot was divided into 3 different areas, Lateral, Central and Medial. The time of initiation of loading for each of these masks was then determined. Reliability of the loading initiation times was determined using type (2,1) intraclass correlation coefficients. Differences between the three forefoot areas were determined using a one-way, repeated measuresANOVA and Tukey's post hoc analysis, if indicated. An alpha level of 0.05 was used for all tests of statistical significance. RESULTS:: The ICC values for the initiation of loading in the forefoot were in excess of.936. The result of theANOVA test revealed that the medial forefoot is loaded significantly earlier with a RIGID foot orthotic compared to either the SHOE ONLY or SOFT foot orthotic condition. This earlier loading caused a reversal of the normal loading sequence of plantar pressure in the forefoot during walking. DISCUSSION:: Previous studies have reported that approximately 10 degrees of forefoot movement, independent to hindfoot motion occurs during the loading response of walking. An additional 3 degrees has been shown to occur from the end of the loading response to mid stance. It would appear from the findings of this study that rigid orthoses prevent this normal independent movement between the forefoot and hindfoot, since the rigid orthoses cause earlier loading of the medial forefoot. The soft orthoses permitted the same lateral to medial forefoot loading pattern also observed in barefoot walking. CONCLUSION:: The results of this study indicate that the use of a rigid foot orthosis inside a shoe, alters the timing of forefoot loading during walking. These findings suggest that the use of soft orthoses permit a more normal forefoot to hindfoot movement pattern during the loading phase of walking. PMID- 11415700 TI - Reproducibility test on a children's insole for measuring the dynamic plantar pressure distribution. AB - INTRODUCTION:: This study aimed to establish the reproducibility of a insole measuring system specifically designed for use inside children's shoes. It is important to carry out such an investigation before any measurements are taken in order to ensure that any errors associated with the system are known and quantified. The children's insole had 84 pressure sensors, compared to the adult insoles, with 99 pressure sensors. The sensor technology was essentially the same as used in the adult Pedar system, and incorporated capacitance-based pressure transducers. MATERIALS AND METHOD:: An eight year old boy, height 123 cm, weight 29 kg, was used in the study. Dynamic tests were carried out with the Pedar insole measuring system. The subject walked on a treadmill, at a speed comfortable for him (3 km/h), with the insoles placed between the foot and the shoe. No fixation of the insole to the shoe was used. Seven runs were completed, each with four tests recorded of more than twenty steps. In between each run the insoles were completely removed from the shoe and then replaced. A zero measurement was also taken before the continuation of the next run. Before analysis of the data, twenty steps were selected. The first two steps in each sequence were ignored and the following twenty steps were used in the analysis, ten each for the left and the right foot. Any additional steps recorded were also ignored. The average force for the test data file was initially noted, together with the local pressure for each of the forefoot, midfoot, heel and toe regions, for both the left and the right foot, separately. RESULTS:: The within test data and in-between test data when walking on a treadmill at constant speed was found to be reproducible (F(0.01)) for 6 out of 7 tests for the left and the right foot. The standard deviation of the average force for the left and the right foot was 3%. The 95% confidence intervals for the mean of the peak pressure in the total object was within 6% for the treadmill and free walking data. There was no significant difference between these data. The 95% confidence intervals for the mean of the peak pressure under the big toe was 9% for the treadmill data and 16% for the free walking data. CONCLUSIONS:: These results, together with other published data (McPoil et al. 1995) show that the pedar children's insole system provides accurate and reproducible measurements of the dynamic plantar pressure distribution. The clinician can therefore use this system with confidence as a therapeutic or rehabilitative tool. PMID- 11415701 TI - Perceived comfort and pressure distribution in casual footwear. AB - INTRODUCTION:: Footwear manufacturers view comfort as a basic customer need that is growing in importance and is increasingly competing with fashion as a reason for buying shoes. A number of researchers have suggested that pressures and forces between the foot and the insole may influence perceptions of comfort (Chen et al., 1994). To date, no study has attempted to measure the relationship between pressure variables and perceived comfort in a range of commercially available footwear and, furthermore, little attention had been given to the dorsum of the foot with respect to comfort and pressure distribution. METHODS:: Twenty male subjects without foot pathology walked the length of a 10 m walkway at a self-selected pace. A 'perception of comfort' questionnaire was designed to measure perceived plantar comfort in six regions (rearfoot medial, rearfoot lateral, midfoot medial, midfoot lateral, forefoot medial and forefoot lateral) and perceived dorsal comfort in two regions (flex-line and lacing). Ten commercially available shoes were selected to represent two distinct groups in terms of perceived comfort: comfortable and uncomfortable. Plantar and dorsal pressure data were collected using a Pedar in-shoe system and Mikro-EMED system (Novel GmbH) respectively. Analysis of variance tests were used to determine differences (p<0.01) between the two shoe groups. RESULTS:: Peak Pressure was found to be significantly greater for the uncomfortable group both for the total plantar surface and for the all regions of the foot measured. Maximum force was significantly greater for the uncomfortable group for the rearfoot and forefoot regions but significantly lower for the midfoot regions of the plantar surface. For the dorsal surface maximum force was found to be significantly greater for the uncomfortable group. Contact area was significantly greater for the comfortable group in the midfoot regions of the plantar surface but was significantly lower for the comfortable group at the dorsal surface. DISCUSSION:: This study showed that perceived comfort at the plantar and dorsal surfaces could be related to peak pressure and maximum force in all regions and contact area in the midfoot region of the plantar surface and the total dorsal surface. The relationship between perceived comfort and peak pressure was consistently negative across the entire surface of the foot, whereas the relationship between maximum force and perceived comfort varied depending on foot region. This emphasises the need for researchers to address specific regions of the shoe when measuring comfort. The findings of this study suggest that pressure measurement systems could be used by footwear manufacturers' attempting to improve the comfort of their footwear. PMID- 11415702 TI - The influence of a small insert, in the footbed of a shoe, upon plantar pressure distribution. AB - INTRODUCTION:: A recent development in plantar pressure distribution research, has been the study of the effects of sensory input on pressure distribution. It has been suggested that proprioceptive and exteroceptive information received from the plantar surface of the foot plays an important role in adapting to high pressures in shoes. Robbins and Gouw (1991) suggested that surface irregularities should be added to the insoles of running shoes to gain correct sensory input. Hayda et al. (1994) found that placing a pad proximal to the metatarsal heads produced significant reductions in forefoot plantar pressures around the first and second metatarsal heads. A development by Villeneuve (1993), 'La Posteropodle', utilized a small insert to maintain postural equilibrium, by stimulating the mechanoreceptors in the plantar surface of the foot. The aim of this study was to measure changes in plantar pressure distribution using a small circular insert. METHODS:: Ten non-pathological male subjects were tested whilst walking, after one day of wearing a pair of oxfords (hard) and running shoes (soft), containing an insert of 4 mm in height placed on a 0.8 mm EVA insole. The foot was split into five sections: (1) midfoot, (2) first metatarsal head, (3) 2nd and 3rd metatarsal heads, (4) 4th and 5th metatarsal heads, (5) the phalanges. A PEDAR system (Novel GmbH) was used to collect in-shoe plantar pressure data, with data collections at the beginning and end of a working day. Subjects were tested under two conditions: (1) the insert 5 mm proximal to the metatarsal heads, between the 2nd and 3rd heads, (2) a control, with no insert. RESULTS:: Preliminary results indicate that whilst wearing a hard shoe the insert had the effect of shifting peak pressures from the first metatarsal head, to the area of the second and third metatarsal heads. Peak pressures were found to be lower with the insert present. This has not yet been tested for significance. With the running shoe there appeared to be no significant differences between conditions with and without the insert. There were also no differences between the beginning and end of the day, for both shoe types. DISCUSSION:: From the results it appears that the insert is successful in both shifting peak pressures from the medial to the lateral forefoot, whilst reducing peak pressures simultaneously. This was only evident in the hard shoe condition however, suggesting that the footbed of the running shoe was perhaps too soft to allow the insert to influence sensory input sufficiently. These findings indicate that there may be implications for the use of small orthotics. Further study is required, however, to fully substantiate this hypothesis. PMID- 11415703 TI - Comparison of two insole materials using subjective parameters and pedobarography (pedar-system). AB - INTRODUCTION:: The objective of this trial was to investigate two commonly used insole materials prescribed for shock-absorption and cushioning concerning subjective and pedobarographic parameters. The design was prospective, controlled, randomized and single-blinded. MATERIAL AND METHODS:: A convenient sample of six healthy male adults without any history of leg or foot injury or pain wore -- in random order -- the custom-made insoles for one week. For both insoles the same base material (TEPEFON(R)) was used, insole 1 was covered with PLASTAZOTE(R) I, 3 mm, insole 2 with PPT, 3 mm. Both insoles had a metatarsal pad. After one week of wearing, in-shoe plantar pressures, measured at the same time of day and within the same type of indoor tennis shoes (sockliner removed), were obtained using the PEDAR-System(R) (Novel GmbH, Munich, Germany). Before each measurement the PEDAR-insoles were calibrated and the subject walked around for 5 minutes to get aquainted with the device. Three trials were performed for three different conditions and average values were determined: PEDAR-insole alone, with PLASTAZOTE(R) insole, with PPT(R) insole. Data were collected at self selected speed, gait velocity was determined using two optical switches on a 10 m walkway. Pressures were normalized to body weight. Main outcome parameters were 'Maximal Peak Pressure' (MPP) and 'Pressure Time Integral' (PTI). These parameters were determined for the whole footsole, medial and lateral heel, medial and lateral midfoot, medial, middle and lateral forefoot, hallux and toes II-V. Additionally the subjects filled in a questionnaire including: time wearing the insoles daily (in hours), sweating (visual analogue scale -- VAS), wearing comfort (VAS), perceived discomfort (location). Three months after the trial they were asked via telephone call whether they were still using the insoles. RESULTS:: 1. The overall MPP and PTI values did not differ significantly between the PEDAR-insole alone and the investigated inserts. There was a tendency for both insoles towards lower MPP and PTI values in all regions except for the toes, especially for the PPT(R) insole. 2. The questionaire showed a significantly higher wearing comfort for the PPT(R) insole in contrast to the PLASTAZOTE(R) insole, though all subjects sweated more with the PPT(R) material. Four of the six subjects experienced discomfort due to the metatarsal pad within the PLASTAZOTE(R) insole, only one within the PPT(R) insole. 3. None of the subjects continued to use the PLASTAZOTE(R) insole, but three continued to use the PPT(R) insoles. DISCUSSION:: Despite the significant differences in the subjective parameters, especially wearing comfort, no statistical significant difference for the overall MPP and PTI values between the PEDAR-insole alone and the investigated insoles or in between the insoles tested could be obtained. Yet there was a tendency for both insoles to lower MPP and PTI values in all regions except for the toes. This might be due to the thickness of the insoles and the reduced space within the toe box. The subjectively better tolerated PPT(R) insole tended to lower MPP and PTI more than the PLASTAZOTE(R) insole. CONCLUSION:: Wearing comfort and pedobarographic outcome measurements did not correlate significantly in this trial. Yet there was a tendency for the subjectively better tolerated PPT(R) insole to lower MPP and PTI more. PMID- 11415704 TI - In-shoe force measurements from locomotion in simulated zero gravity during parabolic flight. AB - INTRODUCTION:: No effective countermeasure for space-induced bone loss has yet been identified. It has been hypothesized that an effective exercise regimen would elicit loads on the lower extremity which resemble those encountered on Earth. Although a treadmill has been used on shuttle flights, the loads to which the lower extremity was exposed have not yet been quantified. It is believed that these loads are much less than the loads experienced in 1G. The purpose of this study was to determine the magnitude of lower extremity loading during tethered treadmill exercise in a 0G environment. METHODS:: Data were collected on five subjects (avg. ht. 177.3+/-10.1 cm, avg. mass 78.3+/-18.0 kg) onboard the KC-135, a NASA airplane used to simulate periods of zero gravity through parabolic flight. Subjects ambulated at 4 speeds: a walk (1.56m/sec), fast walk (2.0m/sec) slow jog (2.75m/sec), and jog (3.35m/sec) on the NASA treadmill operated in either a passive or motorized mode. Each subject wore a harness connected to the Subject Load Device (SLD) to tether them to the treadmill. The tension in the SLD was subjectively adjusted for comfort by each subject. Force data were collected at 60 Hz using Pedar insoles. The number of parabolas per subject was variable due to motion sickness and hardware problems. RESULTS:: Analysis of the insole data showed that the average SLD load was only 35.2% BW, although the values ranged from 20.1% to 56.6%. Maximum ground reaction force values increased with increasing speed and were not affected by treadmill mode. The impulse was higher during walking with the treadmill in the passive mode than in the active mode, but this difference diminished with increasing speed. Subjects tended to run on their forefeet, as shown from the extremely small heel impulse values. At higher speeds, heel contact was absent, while forefoot impulse became more pronounced. DISCUSSION:: All force values were lower than those reported from 1G studies, where typical peak ground reaction forces are 1.2xBW and 2.5xBW for walking and running, respectively. At every speed, the ratio of the rearfoot to forefoot impulse was much lower than reported from 1G studies, and this ratio decreased with increasing speed. CONCLUSIONS:: If the exposure to forces similar to those in 1G is a requirement for countermeasures against space-induced osteoporosis, the loads in the SLD must be greatly increased and should be directly measured before exercise. PMID- 11415705 TI - Transformation and average as handy tools for pressure measurement data analysis. AB - INTRODUCTION:: Pressure measurement data underneath[Table: see text] the foot represent usually a large number of data files which is equal to double product of the number of the subjects in study and of the number of the trials for each foot. Besides, 'normal' parameters are necessary to compare the results for the controls and for the subjects in study. Statistical parameters (mean values and standard deviations) are not very convenient when it is required to establish cartographical and geometrical parameters or peak pressures, maximum forces, contact areas, contact time and others for foot typing or as a standard. Sometimes it is reasonable to obtain average pressure data as a base for further data analysis. But taking into account that pressure distribution pattern may differ by its length, width and angle of the inclination of the foot axis to the horizontal it was necessary to solve a problem of transformation of any pressure picture to the given length and width (for vertical location of the foot axis). METHODS:: The transformed coordinates X(*) and Y(*) are defined from the following system of equations: X = Y(*) x width/width(*) x sina + X(*) x length/length(*) x cosa + X(o) Y = Y(*) x width/width(*) x cosa - X(*) x length/length(*) x sina + Y(o) where X(o), Y(o) -- the coordinates of the origin; width, length, a -- parameters of the initial pressure picture; width(*), length(*) -- parameters of the transformed pressure picture. X, Y -- are real values but the values of pressure are defined only in the integer coordinates. One of the main parts of average is a reduction of the time process to any number of frames. If N(max) is a maximum number of frames in one of the data files than all other data files must be transformed with accordance to the following formulas: for j-th frame the value of pressure for sensor with coordinates x, y is: P(*1)(xy) (j) = P(1)(xy) (i) x ki + P(1)(xy) (i+1) x k(i+1), where P(1)(xy) (i) -- pressure for sensor with coordinates x, y of the 1-th initial time process; j = 0, ellipsis, N(max) -1; j-th frame corresponds to i-th frame in the initial time process as i = integer(j) x (N -1)/(N(max) -1), N -- a number of frames in initial time process; coefficients k(i) and k(i+1) are equal to k(i) = j x (N -1)/(N(max) -1) -- I, k(i+1) = 1 - ki. Then the values of pressure are averaged: P(*)(xy) (j) = 1 / L x S P(1)(xy) (j), where L is a number of data files. Corresponding software was developed. RESULTS AND DISCUSSION:: 38 files of pressure measurement data of the teenagers from military school were chosen for transformation and average. Statistical errors of transformation and average were calculated for each data file (for peak pressure, force and area) and for all parameters from two software packages: Novel-win v.95.121 and Novel-ortho v.95.121. The maximum errors are for peak pressures (about 7%-10%) and contact areas (about 6%). CONCLUSION:: Transformation and average programs can be successfully used for data analysis additionally to traditional statistical software: groupmask evaluation program. PMID- 11415706 TI - The adaptation of the foot to heavy loads: plantar foot pressures study. AB - INTRODUCTION:: The foot serves as the main shock absorber during various activities as walking running and jumping. There are static and dynamic mechanisms which play a role in the adaptation of the foot to weight bearing walking with weights. The function of the foot while weight bearing was studied mainly under static conditions. We designed an experiment to explore dynamically the adaptation of the foot to weight bearing walking with weights. METHODS:: Ten healthy subjects participated the study. They walked barefoot over an EMED force plate which based on multiple pressure capacitance sensors. Each subject walked three times. The first walk was without any weight, the second walk with weight of 20 kg in backpack and the third walk with weight of 40 kg. Contact area, peak pressures, peak forces and duration of contact time were measured in seven areas of interest of the foot-heel, midfoot, lateral, central and medial forefoot, lateral toes and hallux. RESULTS:: The pressure time integral and force time integral increased in most areas of the foot except for the midfoot in the 20 kg walk. The largest increase was at the central and medial forefoot. Adding additional 20 kg to the load did not increased the loads on the midfoot but increased further the loads mainly at the central and medial forefoot. CONCLUSIONS:: The human foot adapts itself under loading condition by maintaining the medial longitudinal arch. Increasing the loading further activates compensatory mechanisms which maintain the longitudinal arch and shifts the the loads to the central and medial forefoot. PMID- 11415707 TI - Velocity of the cop as a reliable parameter for early diagnosis of coxarthrosis. AB - INTRODUCTION:: The first symptoms of the degenerative disease of hip joint appear very early when people are about 14 years old. But clinical and X-ray characters manifest themselves insufficiently: temporary slight pains in hip joints on loading, insignificant painfulness along posterior surface of the joint during palpation, slight lameness on the diseased side. The purpose of this study was to try to involve gait analysis in the clinical research of the teenagers with coxarthrosis and to define the parameters which may be used for early diagnosis of coxarthrosis. METHODS:: 38 male military school teen-agers which have no history in the foot disorders and 20 male and 9 female with coxarthrosis were used for study. Plantar pressure distribution measurements were performed with the mini-emed system (Novel, Munich, Germany). Five dynamic records of each foot were made on the pressure platform (28 x 60 sensors, resolution -- 3 sensors/cm(2), sampling frequency -- 16 Hz). Three trials of gait were accomplished by each subject with inserted insoles (99 sensors in each insole) at the distance about 10 m. Novel-win v.95.121 and Novel-ortho v.95.121 packages were used for data analysis. RESULTS:: For the teenagers with coxarthrosis the loading of heel area is significantly less (< = 15 N/cm(2)), maximum force is generally less. Contact time (in percent of roll-over-process) in the forefoot and toes is increased and is equal to 100% very often. The most interesting results are obtained for the curves of the velocity of the centres of pressure versus time. For the controls this curve has one or two maxima (during 50%-70% of roll-over-process when heel, midfoot and sometimes forefoot are loaded), then a short period of insignificant changes of the velocity (up to 88%-94% of roll-over process when only forefoot and toes are loaded) and last a very short period of its increase (toe-off phase). For the most part of the teenagers with coxarthrosis this curve seems to be monotonous, oscillating near its mean value line. DISCUSSION:: These disturbances may be explained by valgus deviation of the neck of femur and insufficient function of the muscles of buttocks as a consequence of its lever web decrease. The muscle forces (particularly abducted muscles) are decreased (about 10%-15%). These data are consistent with previous reported findings. CONCLUSION:: To compensate for the painfulness in the diseased hip joint the teenagers load rearfoot and forefoot less than in the 'normal' foot, increasing contact time in the forefoot. As a consequence of disturbances in hip joint a pathway of the centre of pressure is longer than in 'norm' and its velocity changes oscillating near the mean value when for 'normal' teenagers three definite intervals of its change are viewed. Therefore the first symptoms of displastic coxarthrosis of teenagers may be revealed from plantar pressure measurements by analysing the character of time dependence the velocity of the centres of pressure and loading of different areas of the foot (force and contact time). Additionally, functional state of hip joint muscles, X-ray and clinical symptoms, heat radiation and laser biophotometry data give an opportunity to define the character of locomotor function disorders. PMID- 11415708 TI - Structural and functional aspects of the diabetic foot. AB - INTRODUCTION:: It has been well documented that, on average, patients with diabetes mellitus (DM) have higher plantar pressures than persons without DM, and there are several hypotheses why this may be -- many focused on the role of peripheral neuropathy. The purposes of this study were: (A) To identify and quantify structural differences between age-matched diabetic and non diabetic subjects, and (B) to develop predictive equations for peak pressure in these same patients. METHODS:: Standardized lateral and dorsi-plantar weight bearing plain radiographs of the right foot and ankle of 50 symptom free (NDM), 32 diabetic with no signs of neuropathy (NNP), and 73 diabetic neuropathic (NP) subjects were taken by a single radiographer. Twenty six angular and linear measurements were then made from the films. Plantar pressure distribution from three first step walking trials were collected and peak pressures at 10 discrete sites were determined for each step, and averaged. A one-way ANOVA with Tukey post hoc tests was conducted to test for differences between the means of each of the 26 radiographic measurements at the 0.01 significant level. Regression analysis was also used to predict the ln(pressure) under the heel, midfoot, mth1, mth2, and mth5 regions from the radiographic measurements, range of motion at the talocrural and first metatarsophalangeal joints and weight, for each of the 3 groups independently. RESULTS:: Significant differences were found among the three groups in 5 of the 26 radiographic measurements (Table 1). These included the thickness of the first, second and third metatarsals from the AP view (MT1, MT2 and MT3 -- generally > in NP), and the sesamoid height (NP > others) and fifth metatarsal head height (NDM < others). In addition, non significant trends were found in 4 other radiographic measurements. Regression analysis identified groups of up to five[Table: see text] predictors which accounted for between 27% and 62% of the variance in peak plantar pressure in different subject groups and foot regions. DISCUSSION AND CONCLUSIONS:: We conclude that objective differences in foot structure are present in diabetic neuropathic subjects, and that DM per se is apparently not a factor in these differences. We also concluded that static foot structure accounts for anywhere between approximately 30% to 60% of the variance in regional peak pressure, depending on the region investigated. In most cases predictions were higher in the midfoot and forefoot. PMID- 11415709 TI - The influence of bandage characteristics and inter-individual application variations on underneath bandage pressures. AB - INTRODUCTION:: Cohesive bandages are applied to the legs of racehorses and horses with limb injuries for protection (prevention of abrasion) and support (reduction of movement at the fetlock joint). The support capacity of all commercially available bandages has been questioned. Consequently, the protection aspect of bandaging and the negative side effects, which can be caused by bandaging (eg pressure induced ischemia with subsequent necrosis), were emphasized. High pressures underneath bandages were shown to cause reduced blood flow. Pressures underneath certain types of bandages were shown to be higher than under others. It is unclear if these differences were due to differences in material characteristics between the bandage types or caused by differences in application by the trainers. The purpose of this study was to determine (a) if differences between different types of bandages are observed if these are applied similarly and (b) if earlier observed differences in pressures underneath bandages are reproducible when wrapped by different trainers. METHODS AND MATERIAL:: (a) A wrapping machine for the application of bandages to an artificial joint (simulating the human knee joint) was designed and built. The machine allows to wrap bandages with adjustable, constant tension under well defined wrapping angles in order to simulate a wrapping technique similar to the wrapping by trainers. The artificial joint is equipped with two pressure sensors (Parotec, Germany). After bandage application, the artificial joint was placed in a specially designed testing machine and cycled through 200 cycles from 0 degrees to 90 degrees of flexion at a frequency of 0.5 Hz. Pressure data were collected at a frequency of 20 Hz. The force required to unwind the bandage from its core was measured with a material testing machine. Five samples each of four different 4" wide cohesive bandages were tested: 'E': high modulus latex, 'V': low mod. latex, 'C': low mod. latex, 'F': medium mod. laminate non-latex. All bandages were applied at manufacturer suggested tensions (50% intercept length). (b) Fifth three trainers from the Kentucky Horse Center (Lexington, KY) and the Payson Park Training Facility (Indiantown, FL) volunteered for the study. They applied their favoured brand and a type 'F' bandage to an artificial horse limb. Underneath bandage pressures were collected using a pressure mat (Mikro Emed, Novel GmbH, Germany) placed over the fetlock joint. RESULTS AND DISCUSSION:: (a) Maximum pressures during maximum flexion underneath the type 'E' bandage (5.7+/-0.4 N/cm(2)) were significantly higher than for all other bandages (no difference between other bandages, range 3.5+/-0.6 to 4.0+/-0.4 N/cm). The unwinding force for the type 'F' bandage was significantly lower (10.1+/-5.5N) than for all other bandages (range 22.7+/-11.3 to 42.3+/-15.1N). (b) Trainers applied the type 'E' bandage significantly tighter than the other bandages (18.7+/-3.7 N/cm(2) vs a range of 7.5+/-5.4 to 10.4+/-6.1 N/cm(2)); a tendency was observed that the type 'F' bandage was applied looser than the type 'V' bandage. The results of part (b) of this study are consistent with the earlier study. Based on the results of part (a) of this study it can be speculated that (I) The high pressures underneath the type 'E' bandage are clearly due to its material characteristics, and (II) The differences between the other bandages are not caused by material characteristics but probably by the differences in unwinding force. If the unwinding force is higher than the force required to extend the bandage to 50% intercept length, trainers will probably wrap tighter as suggested and desired. PMID- 11415710 TI - Load weight determination during dynamic working procedures using the pedar foot pressure distribution measuring system. AB - INTRODUCTION:: In 1994 a new BIA research project was launched, which deals with the development of a portable measuring system for recording of external stress parameters (e.g. body postures and handled load weights) at different places of employment. The investigations are carried out against the background of a newly recognized occupational disease (since 1993), which is that of damage of the spine following long-term repetitive lifting and carrying of heavy loads as well as working in an extreme trunk bending posture. As a part of the study it should be clarified if the worker's handled load weight can be determined by using a foot pressure distribution measuring system. METHODS:: In the case of a static working posture the handled load weight can simply be calculated by taking the difference of the measured weight and the known body rest weight of the worker. In dynamic situations the measured total foot reaction force consists of a static and a dynamic component, which is caused by the body movement and which is normally bigger than the load gravitational force. For load weight determination it is important to separate the dynamic forces from the measured total force to get an idea about its gravitational rest part. Therefore the body dynamics are measured synchronously to the foot pressure by using angle and angular velocity sensors. The herewith received data are used as input values for a developed biomechanical link segment model, with which it is possible to predict the foot reaction force. So even in dynamic situations an estimation of the handled load weight can be made by comparison of the predicted with the measured foot reaction force. RESULTS AND DISCUSSION:: In laboratory tests the load weight could be determinated reliably for different working processes over a total measuring time of 40 seconds. During the measurements the Pedar insoles were placed directly under the foot so that the accuracy of the static force measurement was reasonably high (with a maximal deviation of 5% to the real force). When the insoles were put in shoes, additional forces resulting from the bent shoe leather complicate the load weight determination. This effect still has to be analysed to compensate for it in future measurements. CONCLUSION:: The laboratory results encourage us for future measurements at places of employment. Currently our work focuses on an increase of the total measuring time and on a modification of the measuring system to a portable system without any connection to a computer. A field study is in preparation. PMID- 11415711 TI - Impact forces upon landing single, double, and triple revolution jumps in figure skaters. AB - INTRODUCTION:: Competitive success in the sport of figure skating has been largely attributed to the skater's ability to execute multi-revolution jumps, landing backwards, on a single leg, in a prescribed configuration. The extent to which additional revolutions contributes to greater vertical force upon landing was assessed to determine if advanced technical feats places these young athletes at risk of potential impact related injuries. METHODS:: Twenty four competitive figure skaters ranging in technical classification (Juvenile, Novice, and Senior) and age (10 to 26 yrs.) respectively, gave their informed consent to participate in the study. Skaters were asked to execute single, double, and triple revolution jumps to the best of their ability. Landing impact data between the plantar surface of the foot and the insole of the skate was quantified with the use of the Micro EMED Insole System (Novel GmbH, Munich, Germany). The insole was placed inside the skate of the landing foot, tethered to the leg with the data acquisition unit secured to the lower back of the subject. In addition, all trials were video taped by four Panasonic cameras. Three dimensional video analyses were later performed using the Ariel Performance Analysis System and used to co-ordinate the impact data with the technical aspects of the jumps. The EMED Extern -- Multimask Software Package was employed to process the Micro EMED data. The foot was divided into two regions for data analysis: rearfoot (0-50% of the length of the pressure print) and forefoot (50-100% of the length of the pressure print). Impact forces and foot pressure data were then analysed and the results documented for the following variables: Peak impact force (%BW), Peak pressure (N/cm(2)), Pressure time integral (Impulse-N/cm(2)), Force time integral (Impact-%BW) and measures of the Peak force-time differences between the fore and rearfoot (ms). A repeated measures analysis of variance was performed to determine significant differences among the above listed variables, between jumps and between groups as stipulated by their technical classifications (p = 0.05). RESULTS:: ANOVA results indicated that gender differences were not significant for any of the examined variables, thus gender groups were collapsed for further analysis. Statistically significant differences in peak impact force (%BW) and time sequencing of foot contacts (ms) were revealed both between jumps and groups (p < 0.05). An analysis of effect size examined the magnitude of the differences in impact force and force-time differences of triple revolution jumps. A range of moderate (0.5) to large (1.3) differences in relative force and force-time differences between single, double, and triple jumps and technical classifications were detected. Furthermore, an inverse relationship was found between relative force (N/kg) and time differences between the fore and rearfoot peak force contacts (m/s). DISCUSSION:: The relationship between relative impact force and time delay between fore and rearfoot contacts suggests that the time taken to distribute the force of triple revolution jumps is significantly less than the singles and doubles. Although jump height was not quantified as a primary purpose of this investigation, subsequent kinematic analyses revealed that jump height was not significantly different among single, double, and triple revolution jumps. Therefore, increased height is not suggested as possible cause of higher impact force or shorter footfall time. However, it is suggested that the increased time required for rotation places the skater closer to the ice at time of impact. A 'collision type' landing seems to be a result of higher revolution jumps. Decreasing the time to properly dissipate the force may also suggest that the impact propagated through the lower extremity musculo-skeletal system is of greater magnitude and intensity introducing greater potential for injury. PMID- 11415712 TI - Ground reaction force and plantar pressure reduction in an incremental weight bearing system. AB - INTRODUCTION:: With increasing frequency, harness-supported treadmill ambulation has been advocated in patient use in order to provide reduction in weight-bearing to healing tissues and as a method which reduces the energy cost of treadmill ambulation. The purpose of this study was to analyse the ability of one of these devices (Zuni Exercise System) to support a percentage of a subject's body weight during walking and running and to explore the relationship of unloading to pressure reduction in selected plantar surface regions of the foot. METHODS:: Ten healthy male volunteers with no known foot pathology participated in this study. In-shoe plantar pressure (PEDAR) and vertical ground reaction force (GRF) were measured during walking and running at full body weight and at a 20% body[Figure: see text][Figure: see text] weight supported setting. Statistical evaluation used a one way ANOVA and a post-hoc paired t-test with significance set at p < 0.05. RESULTS:: Walking with a setting of 20% body weight supported was achieved with a reduction of the first and second vertical force peaks of 23.8+/-7.3% and 27.2+/ 4.1% respectively, somewhat greater than the selected setting (Figure 1). The total force time integral during walking unloaded was 22.8+/-3.3%, which was only slightly greater than the selected 20% reduction. During running the active vertical force peak and total force time integral were reduced by 19.9+/-6.0% and 20.0+/-3.3% respectively during the unloaded condition (Figure 2). Plantar pressures were reduced from 6.8 - 27.8% during body weight supported conditions. The reduction in plantar pressure was variable across different regions of the foot with the toes in walking and the medial forefoot region in running being the least unloaded. DISCUSSION AND CONCLUSIONS:: The Zuni Exercise System appears to be a valid device to predictably reduce the vertical component of the GRF during walking and running with 20% body weight supported. Plantar pressures were reduced during body weight supported conditions but the reduction varied at different regions of the foot. However, the variability of the reduction across subjects was substantial, implying that a given regional pressure may not be as predictably reduced during a body weight supported condition. This highlights the difficulty a practitioner has in the ability to confidently reduce pressure by a set amount in a selected foot region by using assistive devices without direct in shoe measurements. PMID- 11415713 TI - Chinrest pressure in violin playing: type of music, chin rest, and shoulder pad as possible mediators. AB - INTRODUCTION:: This study was conducted to determine if there are differences in pressure on a violin chin rest during performance due to changes in shoulder pads, chin rests, or pieces of music performed. It was also hypothesized that these differences would correlate with anthropometric measurements made of the subjects' necks and shoulders. METHODS::: A Novel EMED pad was used over the chin rest, containing 120 sensors, each one centimetre squared. The sampling rate was 50 Hz. A total of ten subjects, all professional violinists, performed excerpts from a Bruch violin concerto and a Handel sonata. Anthropometric measurements were made of the subjects' necks and shoulders. Each subject began by using his own violin with his usual shoulder pad and chin rest set-up, and then with the author's violin. Two different shoulder pads and three different chin rests were placed on the violin, in all possible combinations, for both pieces of music. This resulted in a total of fourteen conditions given in randomized order for each subject. Subjects were asked to give verbal feedback regarding the comfort and ease of playing of each shoulder pad and chin rest. Data was collected for three trials of each condition, and a mean for the three trials was obtained for each dependent variable. The five dependent variables were: peak pressure, maximum force, pressure/time integral and force/time integral, and total contact area. Intraclass coefficients of reliability were calculated for the dependent variables. Pearson's correlation coefficients were computed to determine if there were significant relationships between the five dependent variables and violin height, shoulder width, and neck length. RESULTS:: Pressure/time integral achieved the highest reliability with a coefficient of 0.9310, followed by peak pressure at 0.9152, force/time integral at 0.8817, maximum force at 0.8685, and total area at 0.8571. Multivariate analysis of variance detected significant differences in levels of pressure and force for the different conditions. Subsequent one-way ANOVAS andt -tests determined significantly less pressure on the Wolf chinrest as compared to the other chinrests. There were also reliable differences in pressure between the two pieces of music, with the Handel sonata excerpt showing significantly less pressure. There were no differences detected between the two shoulder pads. Significant relationships between neck length and shoulder width, and peak pressure, pressure/time integral, maximum force, force/time integral, and total contact area were found. A discussion of these results, and conclusions and implications for further study, are currently in progress. PMID- 11415714 TI - Coupling force distribution and finite element model for calculating the consequences of distributed force input. AB - INTRODUCTION:: The aim of the study is to repeat an important step in the development of biomechanics in relation to pressure distribution measurements. Just one decade ago (a) one channel force measurement and (b) 'one channel models' were developed to a certain level of sophistication so that the force could be used by the model to calculate movement and load of the body. In contrast, work in the field of pressure distribution was focused to (a) multi channel measurement, but (b) no models were used to calculate the consequences of the forces measured. In the following, a multi-channel Finite Element Model (FEM) is introduced for that purpose. The body studied is a mattress; the results attained are helpful under two aspects. Firstly, the FEM provides information on the stress inside the mattress; this aspect is of interest for nearly all bodies. The second aspect is a special problem of mattresses and the like: as the user covers the surface of the mattress, there is no direct information on the shape of mattress and the human body, and measuring procedures for the shape are quite difficult and coarse. METHODS:: The mattress is modelled as a set of a(*)b(*)c vertical springs where c springs are standing on top of each other. The springs are interconnected in a horizontal direction. On top of the set an extra layer with very strong horizontal coupling is placed, which is the model of the cover fabric. Pressure distribution measurements showed that the fabric is responsible for the so called 'hammock effect' which significantly reduces the quality of the mattress. a(*)b forces are input into the model; each force influences all springs because of the horizontal coupling. The distance of the springs is designed according to the distance of the force sensors, which in turn reflect the importance of the mattress area studied. In the trunk area the springs are spaced 15(*)30 mm, in the remaining mattress area the spacing is 72.5(*)43 mm; two springs are standing on top of each other. Each spring defines a volume element, the corner points of each element are connected to the neighbouring element, transferring stress and strain this way. The constraints are specific for the mattress type. APPLICATION:: Utilizing this model and a software package I-DEAS installed on a work station, stress and displacement to three axes of each volume are calculated, moreover the shape of the mattress surface and thus the shape of the body lying on it are attained. The standard measuring procedure is as follows: 1. A mattress is loaded by boards and balls so that the shape of the surface under pressure is known; from pressure distribution measurements the model calculates the spring data. 2. Various subjects are placed on the mattress and measured; the model calculates the surface shape of their body utilizing the spring data. 3. Data of lying quality (concerning comfort and support) are assessed from the body shape and recommendations concerning improvement of the mattress are derived. PMID- 11415715 TI - Finite element modelling of plantar pressure beneath the second ray with flexor muscle loading. AB - INTRODUCTION:: Little is understood about the effects of flexor loading on plantar pressure distribution. The goal of the current work is to model flexor muscle loading applied to the distal phalanges in order to study the effect of these loads on plantar normal stress (pressure) beneath the metatarsal head. METHODS:: The finite element model is a two-dimensional, plane strain sagittal section incorporating the second metatarsal, proximal phalanx, and plantar and dorsal soft tissue (Figure 1). The metatarsophalangeal joint is simulated by a nodal hinge that transfers loads and produces reasonable kinematic motion between the articular surfaces of the proximal[Figure: see text] phalanx and metatarsal head. Soft tissues are simulated by a uniform continuum. A single flexor tendon passes over the condyle of the metatarsal heads with sliding contact against intervening soft tissue, and is attached to the distal end of the proximal phalanx. A rigid element at the proximal end is fixed by boundary conditions to simulate reactions at the distal cuneiform joint. Material properties of bone are from published values, one tenth the stiffness of bone is used for the flexor tendon, and the soft tissue continuum is hyperelastic using coefficients obtained from compression of the heel plantar fat pad. A 188 N vertical ground reaction force and a flexor tendon load at a 10 degree angle from the X (horizontal) axis are applied to the model. RESULTS:: Figure 2 shows Y direction normal stress distribution along the plantar surface for two load cases: no load and a 250 N load to the flexor tendon. DISCUSSION:: Bending moments at the proximal metatarsal correspond to values obtained by Sharkey et al. Tension in the flexor tendon served to counter the moment in the metatarsal created by the vertical load, and at the same time, to apply an additional axial load. Under flexor loading, focal plantar pressure shifts toward the proximal phalanx and yields a 60% reduction in peak pressure, indicative of the load sharing between the sub metatarsal head and subphalangeal regions. [Figure: see text] CONCLUSIONS:: The model yields verifiable and reasonable reactions and a significant relationship between flexor muscle loading and peak plantar pressure. Refinement of the model, such as adding the middle and distal phalanges, should reveal further insight into the mechanics of plantar loading. PMID- 11415716 TI - Analytical approaches to the determination of pressure distribution under a plantar prominence. AB - INTRODUCTION:: The purpose of the current research was to develop an analytical model for the interface between the metatarsal head, surrounding soft tissue, and the shoe. Results of 'peak plantar pressures' computed from this model were compared to previous results obtained from clinical data and from a finite element (FE) model. METHODS:: An analytical model for determining the pressure distribution at the interface between the foot and shoe insole was developed based on the solution[Figure: see text] for a rigid cylinder and a single elastic layer mounted on a rigid half-space originally developed by Meijers (see Figure 1). The rigid cylinder was used to represent the metatarsal head region (including bone and soft tissue). Material properties and loading conditions were chosen to reflect those used by Lemmon et al. for 'Normal' and 'Reduced' soft tissue thickness cases. RESULTS:: Peak pressures computed from the analytical model are plotted against insole thickness in Figure 1, alongside experimental and FE model data from Lemmon et al. Correlations between analytical, FE, and experimental results for both the Normal and Reduced soft tissue cases were all highly significant (r(2) = 0.86, p < 0.01). DISCUSSION:: The analytical model produced unrealistically high peak pressures compared to the FE and experimental results, with differences of approximately one order of magnitude. This was not surprising, given that the entire metatarsal head region (soft tissue included) was modelled as a rigid cylinder. The soft tissue on the bottom of the foot, and the much greater surface area over which the loads are carried in the real case, contribute greatly to reducing these peak pressure values. However, the downward trend in peak pressure with increased insole thickness was retained. CONCLUSION:: It is anticipated that the use of more accurate and detailed models will greatly improve the initial results. PMID- 11415717 TI - Assessment of pedar and F-Scan revisited. AB - INTRODUCTION:: Previous experimental results have suggested that the PEDAR system (Novel Electronics, Inc.) for plantar pressure measurement was able to measure known static pressures more accurately than the F-Scan system (Tekscan, Inc.). These measurements, however, were made prior to two recent developments of the F Scan system (the release of new resistive ink sensor insoles and software allowing for calibration via an air pressure bladder). We sought to address the following questions: (a) Which system measures known pressure more accurately? (b) Which system is more repeatable? (c) Which system has lower measurement variance across a uniform distribution? (d) Is F-Scan repeatability improved with recalibration? METHODS:: Pressure measurements were made using a pair of PEDAR insoles and a pair of new ink, F-Scan insoles. All measurements were made in a Novel air bladder, which provided generated known, uniform pressure distributions. The PEDAR insoles were calibrated using the standard protocol involving the air bladder. This calibration was used for all PEDAR measurements. F-Scan insoles were calibrated on each of three testing days using the new bladder calibration technique. The second testing day was one day after the first, while the third testing day occurred four days following the first. Each set of measurements involved placing both insole pairs (one on top of the other) in the bladder and loading the bladder, at 100 kPa intervals (100 up to 500 kPa and 500 down to 100 kPa). Data were recorded at each interval and the bladder was emptied between each recording. One set of data was collected on the first testing day. On each of the remaining test days, one data set was collected with the F-Scan system using the calibration from the first day, and another set was collected using a calibration obtained that day. For each recording we computed the average and standard deviation of pressure values across sensors in a 3 cm x 3 cm area in the 'forefoot' region. RESULTS:: Across all data sets and all conditions, the mean absolute differences between the area average pressure and the bladder pressure gauge value were 10 and 86 kPa for the PEDAR and F-Scan systems, respectively. The mean absolute differences for the five sessions were 10.7, 5.5, 11.5, 3.3, and 10.5 kPa for the PEDAR insoles. For the F-Scan insoles the means were 110, 104 and 83 kPa with the first day calibration, and 110, 73 and 60 kPa with same day calibrations. The mean standard deviations across the indicated region were 9 and 19 kPa, respectively. DISCUSSION:: The results obtained appear to suggest that the PEDAR insole system still measures uniform pressure more accurately and with greater repeatability than does the F-Scan system with new resistive ink insoles and with calibration via a bladder. The PEDAR system also demonstrated lower variance across sensors, suggesting that sensor to sensor random errors were lower in the PEDAR system. These findings, generally, indicate that applications requiring the greatest accuracy and/or repeatability would be best performed with a PEDAR system. It was noted, however, that repeatability of the F-Scan system was improved when a calibration from a single day was used across days. This finding has beneficial implications for F Scan users in clinical settings, as it would suggest that trends over time could be better detected if the insoles initially used by a patient are not discarded or recalibrated. CONCLUSIONS:: In summary, we must concur with previous reports that the PEDAR system is likely the system of choice when the greatest accuracy and repeatability are desired; however, for some common clinical applications, F Scan users can improve the utility of their data by using the new insoles, calibrating via bladder, and retaining patients' initial insoles and calibration files. PMID- 11415718 TI - Observations on the F-Scan in-shoe pressure measuring system. AB - BACKGROUND:: F-Scan (Tekscan Inc.) is a commercial in-shoe force/pressure measurement system. The system has been used in a number of useful clinical studies focused mainly on the evaluation of various therapeutic measures including footwear and in-shoe orthoses. More recently, the accuracy and reliability of the system has come under closer scrutiny, highlighting the need for further debate supported by experimental studies. We have critically evaluated the performance of the F-Scan sensor and report here on bench test findings examining calibration error, hysteresis and creep, and the accuracy and repeatability of output force measurements. METHOD:: The F-Scan system was used to measure output mass following calibration using the standard protocol across a range of subject determined input masses. A small jig-mounted force meter was used to test groups of individual sensing units (4 @ cm(2)) for within and between sensor accuracy in full size and adjusted sensors. An Instron servohydraulic materials testing unit was used to evaluate creep, hysteresis and the repeatability of output forces over repeated loading cycles for full size and cut sensors. RESULTS:: Average calibration error: 4% Creep: 19% Hysteresis: 21% [Table: see text] CONCLUSION:: 1. The F-Scan calibration protocol is inaccurate. 2. Significant within and between sensor variability in output forces can be expected. 3. The creep and hysteresis properties of the sensor are poor. 4. The F Scan sensor does not yield repeatable measurements. 5. When the sensor is adjusted to a smaller size output is adversely affected. The F-Scan system has a limited capability for absolute accuracy but may be used for quantitative studies provided its limitations are noted. PMID- 11415719 TI - Gait function after fracture-dislocation of the midtarsal and/or tarsometatarsal joints. AB - INTRODUCTION:: Fracture-dislocations of the Chopart or Lisfranc joint line represent rather rare injuries which usually result from high-energy trauma. During the last decade surgical therapy, including open reduction and internal fixation, had been accepted as the best way to achieve an optimum return to function. Repeatedly, it had been stated that anatomical reduction represented the prerequisite for satisfactory results. But nevertheless, despite benign appearances on postoperative radiographs, even disappointing clinical results may occur after this severe type of complex foot injury. In order to get more insight into foot function after Chopart and/or Lisfranc joint trauma and eventually characterize those parameters with most importance for surgical intervention and a non-disturbed gait function, a post-reconstruction study was performed in 25 patients after surgical therapy and definite healing. METHODS:: Twenty five patients were examined clinically according to a standardized protocol 1-8 years after injury and surgical reconstruction and the Maryland Foot Score (100-point rating scale) was calculated from clinical and anamnestic data. Further, standard radiography of the foot was performed. Five patients had suffered from an injury of the midtarsal joints, 11 patients had an isolated injury of the tarsometatarsal joints and 9 patients had a combined injury of both joint complexes. Surgery generally included open reduction and internal fixation employing AO small fragment screws and/or K-wires, sometimes supplemented by an external fixator. Generally, at the time of examination implant removal had been performed, again. Gait function was studied employing an EMED-SF 4 platform integrated into a walkway of 6 m length. Standard parameters were calculated in 9 specific masks employing the Novel-win software. Essentially, an intraindividual comparison with the data from the non-injured extremity was performed. Further, lateral-medial force indices were calculated using Novel-orthopaedics. RESULTS:: Intraindividual comparison of the local impulse distribution pattern correlated considerably well with the total count of the Maryland Foot Score. The mediolateral force index showed that patients with a former lesion of one of the foot columns (medial or lateral) tended to load the non-injured column, mainly. This could also be recognized in patients with apparently good or excellent gait function. The severity of radiographically visible posttraumatic arthrosis obviously did not influence gait function to a major degree, but a loss of length of one of the foot columns following a comminution injury component or a shift of the foot axis either in the horizontal or in the vertical direction demonstrated a substantial influence on gait quality. Due to the limited number of patients, an influence of technical variations of surgical reconstruction could not be demonstrated. CONCLUSION:: As a practical consequence of the presented study it may be concluded that the correct alignment of the foot axes, including a correct length proportion of the medial and lateral foot columns after an injury of the Chopart and/or Lisfranc joints, should represent a major goal of therapy. PMID- 11415720 TI - Foot pressure distribution: methodology and clinical application for children with ankle rheumatoid arthritis. AB - INTRODUCTION:: Foot pressure measurements furnish information about distribution of pressures, forces, time and contact areas under the foot during standing and walking. Foot pressure measurement has been used in a number of rehabilitation and athletic applications in adults, however, little has been published regarding the clinical usefulness of this technology for children with disabilities. Children with juvenile rheumatoid arthritis (JRA) are reported to have various foot deformities and gait deviations and clinicians report that the children may have foot pain such as metatarsalgia. These may be treated with specially fitted shoes, shoe modifications or ankle foot orthoses. The purpose of this preliminary study was to describe the methodology used to quantify foot pressure distribution patterns and, further, to describe the patterns seen in individual children with JRA compared to aged matched typical children without JRA. METHODS:: Pressure, area and force were measured using the EMED-F system including a platform with 2048 capacitive pressure sensors and a computerized data collection and analysis system. Children were asked to walk comfortably and normally across the platform while time, pressure and area measurements were automatically taken. Other data collected were height and weight, observational gait analysis and lower extremity range of motion measurements. Data are reported for the entire foot, as well as particular areas of the foot that are of interest. For this study, eight discrete areas or masks were identified (medial and lateral heel, medial and lateral midfoot, first metatarsal, lateral four metatarsals, great toes and four lateral toes) for description. Information reported for each area and the total foot included force, peak pressure, total area, pressure time integral and force time integral. Data from three pairs of children were analysed and differences were described. RESULTS:: Several differences in the descriptive data were noted and will be highlighted. Children with JRA had striking asymmetries in several variables, higher peak pressures and in increased total foot pressure time integrals and force time integrals. This information is presented to improve our understanding of the patterns under the foot so that (1) appropriate treatment strategies for foot impairments may be better prescribed for children with JRA and (2) to assist in planning for treatment of gait abnormalities. This preliminary work will also form the basis for determining the clinically meaningful variables to consider in a larger study and statistical analysis. PMID- 11415721 TI - Clinical and gait analysis of 171 unilateral calcaneal fractures. AB - INTRODUCTION:: Reviewing the management of calcaneus fractures, conservative and operative treatment are known without a generally accepted, well proven therapeutical strategy or tactics compared to other types of fractures. The purpose of this study is to define the parameters of the differences between the fractured calcaneus and the intact one, as well as the two types of treatments using the gait analysis method. MATERIALS AND METHODS:: Retrospective clinical investigation and gait analysis have been performed on 217 patients with calcaneus fractures. An analysis of the X-rays was also included. The Novel 101 B pedobarography analyser with platform 102H (4 sensor/cm(2)) was used for pressure distribution analysis. The region of the foot was divided into four masks, Mask 1 (M1) the area of the heel, Mask 2 (M2), the mid-foot, Mask 3 (M3), metatarsal region, and Mask 4 (M4) the region of the toes. The total area of the foot (TOT) included all four masks. Eighteen bilateral fractured calcanei have been omitted from the evaluation, owing to the lack of control foot, and 28 patients were excluded for the other complications of the fractures foot, complications of the control foot or complications extending to both feet. The data on the 171 fractures (68 patients treated conservatively and 103 patients treated operatively) of the calcaneus were compared with their control feet. The average age was 52.0 years (+/- 2.7) and the average follow up time was 49.9 months (+/- 17.5). The statistical analysis was performed using SPSS for Windows. Examination of correlation, repeated measurement analysis of variance, ANOVA, and multiple comparisons were performed by Bonferoni. Linear discriminating analysis was also performed. Significance level was defined as p<0.05 in each case. RESULTS:: All investigated parameters (area, max.F, max.P, absolute and relative contact time, FTI, PTI.) of M2 were significantly higher on the fractured side than on the intact one. The investigation of the whole foot showed significant increase concerning the area, and significant decrease in the max. force, FTI and PTI values. The differences between the conservatively treated and the intact foot values, as well as the differences between operatively treated foot and the intact one, demonstrated a significant decrease in the FTI TOT and the PTI TOT in both groups. A significantly higher difference was also demonstrated when the difference in the value of PTI TOT in the operative group was compared to the conservative one. Therefore, the values of the fractured side of the operative group were significantly lower than the values of the intact one, but significantly higher than the values of the conservative group. CONCLUSIONS:: The gait analysis parameters for the calcaneus fractured and the intact sides allowed for separation of the data according to significant differences. The results of the gait analysis comparing the conservative and operative method of the treatment showed that the surgical method was the better choice. PMID- 11415722 TI - Early diagnosis of coxarthrosis (in-shoe plantar pressure measurements). AB - INTRODUCTION:: Plantar pressure measurement during walking adds dynamic functional data to evaluation of gait disorders of teenagers with coxarthrosis. The duration of the complete gait cycle, its phases (stance and swing phase for right and left feet, double support) and phases of roll-over process are found useful in many clinical applications. METHODS:: In-shoe measurements were completed with Pedar system (99 element capacitive insoles from Novel, Munich, Germany) for 38 teenagers from military school (controls) and 29 teenagers with coxarthrosis. The data from 3 - 5 trials for each subject were collected. The distance was about 10 m, no requirements to the speed and step length were established. Special software was developed. RESULTS:: Time parameters for the complete gait cycle, its phases, phases of roll-over-process were calculated both for the controls and the teenagers with coxarthrosis It was found that stance phase and heel contact phase are decreased for the teenagers with coxarthrosis. DISCUSSION:: Decrease of stance phase, insufficient loading of the heel and decrease of heel contact time are a result of attempts to compensate the painfulness in the hip joint during walking. CONCLUSION:: Time parameters of the complete gait cycle and its phases may be useful in early diagnosis of coxarthrosis and evaluation of efficiency of the treatment. PMID- 11415723 TI - Analysis of plantar loading parameters pre and post surgical intervention for hallux vargus. AB - INTRODUCTION:: The purpose of the study was to compare range of motion (ROM) and plantar loading parameters pre and post surgical intervention for hallux valgus (HV). Fifteen female patients volunteered for the study, providing 27 feet with a diagnosis of moderate or severe HV deformity (greater than 26 degrees HV angle). All participants were screened by the same orthopaedic surgeon to ensure that no other foot deformities or neuromuscular diseases were present that would alter lower extremity biomechanics. Prior to surgery, each patient filled out a five part questionnaire on previous medical history, description and location of symptoms, perceived pain and current functional status. Radiographs were taken from the dorsoplantar and oblique aspects with the patient weight bearing. HV and intermeditarsal angle (IM) were measured from the radiographs. Seated talocrural and first metatarsophalangeal (MP) joint active and passive (ROM) were measured with a hand-held goniometer as described by Norkin and White (1985). Plantar loading patterns were assessed using barefoot for the involved limb using the two step method as described by Meyers-Rice et al. (1992). Data was collected using a capacitive pressure measurement platform (EMED SD Pedography Analyzer, Novel GmbH, Munich) and stored for further analysis. The pressure platform consisted of a 32 x 62 sensor matrix with a resolution of 2 sensor/cm(2). The sampling rate of the platform was fixed at 70 Hz and auto triggered upon first contact. Five acceptable trials were collected for each subject. All of the following were conducted again eight weeks after a Chevron-Aikin Osteotomy surgical correction for HV. Each step was analysed by dividing the foot into seven plantar regions: heel (HL), midfoot (MF), medial forefoot (MFF), central forefoot (CFF), lateral forefoot (LFF), medial toe (MT) and lateral toe (LT). The following variables for each region were generated via the Novel-win software: peak force (%BW), peak pressure (Kpa), length of contact (ms), pressure time integral (Kpa(*)s), force time integral (%BW(*)s), instant of peak pressure (ms) and instant of peak force (ms). A repeated measures multivariate analysis of variance was used to detect differences in each loading and ROM measure before and after surgery. Post surgically, patients indicated that they had less pain and a higher rating of functional status (p<0.05). Differences were found in active dorsiflexion, active plantar flexion, and passive plantar flexion of the MP joint (p<0.05). No differences were found in talocrural ROM or MP neutral position. Radiographic measures indicated that HV and IM angles were changed following surgery (p<0.05). The average correction for HV angle was 14.0 degrees and 4.6 degrees for IM angle. No differences in loading were found for the HL, MF, MFF regions post surgically. Force time integrals increased from 15.7 BW(*)s to 18.8 BW(*)s post surgically in the CFF region (p<0.05). The LFF region exhibited a greater peak force (22.12 %BW vs. 27.12 %BW), force time integral (7.14 %BW(*)s vs. 9.04 %BW(*)s), pressure time integral (9.24 N/cm2(*)s vs. 12.10 N/cm2(*)s), instant to peak force (520.73 ms vs. 592.03 ms) and instant to peak pressure (548.40 ms vs. 607.97 ms) post surgically (p<0.05). Peak force and peak pressure decreased from 15.11 %BW to 6.92 %BW and 423.24 Kpa to 158.00 Kpa, respectively post surgically (p<0.05). Force time integrals decreased from 3.90 %BW(*)s to 1.72 %BW(*)s and pressure time integrals decreased from 10.41 Kpa(*)s to 4.40 Kpa(*)s post surgically (p<0.05). Contact time in the LT region increased from 627.57 ms to 575.33 ms post surgically (p<0.05). All pain scale and functional status measures improved eight weeks post surgically. Dorsiflexion ROM for the MP joint were decreased post surgically but were still adequate for gait (Hetherington et al., 1990). Loading variables indicated that, post surgically, the foot was loaded more laterally with less load on the hallux. Further research is needed to assess if the lateral loading pattern continues long term after Chevron-Aikin Osteotomy surgical correction for HV. PMID- 11415724 TI - Measurement of plantar pressure distribution during gait for diagnosis of functional lateral ankle instability. AB - INTRODUCTION:: Chronic functional instability of the lateral ankle may be difficult to distinguish from mechanical instability when radiological stress tests reveal only small ligamentous defects. For decision making whether to surgically reconstruct the ligaments or not, it can be helpful to use additional information on joint and foot function. Therefore, the aim of a prospective study of patients with longstanding chronic ankle instability was to demonstrate that the dynamic measurement of plantar pressure distribution can identify patients with functional ankle instability. [Table: see text] MATERIALS AND METHODS:: Sixty five patients (mean age 24 (4.6 years)) were included. After clinical examination and radiological stress views, plantar pressure patterns were measured during gait using a capacitive platform, the EMED-SF 2-system. Five trials of each foot were documented and the maximum impulses in eight points of the foot (central heel, lateral and medial heel, midfoot, 1st, 2nd, 5th metatarsal head and hallux) were calculated intraindividually and compared with a group of 100 healthy subjects. The medio-lateral loading factor (MLF) as the quotient of the medial and lateral relative impulses indicated the tendency to walk on the lateral edge of the foot. RESULTS:: Based on clinical criteria alone, two comparable groups of patients were separated, 35 with functional instability and 30 with mechanical instability. After collective analyses of the results, the patients with functional instability showed a significantly increased lateral loading of the unstable foot (p=0 0 1), whereas the mechanically unstable group tended to walk more on the medial side of the unstable foot (Table 1). DISCUSSION:: Dynamic measurement of plantar pressure can identify a group of patients walking on the lateral side of the unstable foot when compared with the stable foot. This finding is explained by a deficit of peroneal strength during stance phase based on a proprioceptive defect after trauma. The new application of the EMED-system provides additional information which helps to make decisions about the individual patient with chronic instability. PMID- 11415725 TI - The variability of the centre of pressure pattern. AB - INTRODUCTION:: The centre of pressure pattern (COPP), or force line, has been used to describe abnormal foot movement during gait and as a method to assess foot orthoses effectiveness. The purpose, therefore, of this study was to measure the between-trial variability of the COPP as well as explore possible differences between shoe-floor and within-shoe measurements. METHODS:: Ten healthy individuals from a pool of 110 volunteers were selected for this study based upon their willingness to participate and the presence of a forefoot and/or rearfoot valgus or varus deformity. An EMED-SF pressure platform was used to collect COPP data at the shoe-floor interface. The EMED PEDAR insole pressure system was used to collect COPP data within-the-shoe. Ten trials of COPP were recorded simultaneously using the two systems as the subjects walked over ground in standardized footwear and two different types of foot orthoses. The footwear used consisted of the Reebok Boston Road running shoe. The two types of orthoses studied were a rigid orthotic with forefoot and/or rearfoot posts and soft orthotic without any posts. Quantification of the COPP was achieved by calculating its absolute integral. RESULTS:: The result of a one-way repeated measuresANOVA and subsequent post-hoc analysis showed that the rigid orthoses condition was significantly different from the soft orthoses or no orthosis conditions. Analysis of the between-trial reliability for COPP integral calculation, however, indicated that there was 43.4 to 48.6% variability. The intraclass correlation coefficients were found to be less than 0.504. DISCUSSION:: While the results showed that the within-shoe shoe only condition and the soft orthoses condition caused the greatest decrease in the COPP integral, the COPP integral was extremely inconsistent, whether measured at the shoe-floor interface or within-the-shoe. CONCLUSION:: The results of this investigation indicate that COPP is highly variable when measured either at the shoe-floor interface or within-the-shoe over multiple trials. Based upon these findings, the COPP, as measured by its absolute integral, would be a poor representation of foot movement and, as such, would not be an acceptable method for the assessment of foot orthoses effectiveness. PMID- 11415726 TI - A literature review of low back disorder surveillance measures and risk factors. AB - OBJECTIVE: The goal of this literature review was to gain insight into low back disorder risk factors via a critical examination of the surveillance measures and analysis techniques employed in existing literature. DESIGN: Fifty-seven original articles were evaluated and categorized as a function of their surveillance measures. BACKGROUND: There have been a plethora of articles concerning the causes of low back disorder, yet no specific risk factors are consistently associated with the development of these disorders. It was hypothesized that different low back surveillance measures and variations in risk factor (dependent variable) measurements have led to the inconsistencies in the literature. METHODS: Five low back disorder surveillance measures and five risk factor categories were defined for this review. Each article was classified on several criteria including: surveillance measures, risk factors, statistical methods, population and type of study. Summary statistics were calculated for the percentage of positive findings as a function of surveillance measure and risk factor category. RESULTS: The most consistently defined surveillance measure was incidence of low back disorder, with 82% of those investigating it as claims from medical records or Occupational Safety and Health Administration records. The combination of surveillance measures and risk factor influenced the outcome of investigations. Ninety-one percent of the direct or video methods of measuring exposure risk factor influenced outcome. Psychosocial measures had positive findings in 70% of the studies examining lost time. Finally, statistical methodology was critical in the outcome of these investigations. CONCLUSIONS: The surveillance measure of incidence had more positive findings, with exposure risk factors and the surveillance measures indicating more advanced stages of low back disorder such as lost time had more positive findings with psychosocial risk factors. Thus, as low back disorders progress to disability, the psychosocial risk factors play a more prominent role. RELEVANCE: In order to prevent low back disorders we must first understand the plethora of epidemiological literature. This literature review provides new insight on the critical issues that have contributed to the results of previous research. PMID- 11415727 TI - Intersegmental dynamics of standing from sitting. AB - OBJECTIVE: To compare the magnitude of muscle moments acting on either end of body segments with other motion-dependent and gravitational moments during the task of standing up. DESIGN: A mathematical model which partitions moment components was developed and applied and descriptive analysis of the resultant data conducted. BACKGROUND: It has been hypothesized that segmental interactions may be exploited during the task of standing up from sitting to lessen demands for the production of large muscle forces. There has been no investigation of the relative sizes of these interactions nor of their impact on the task. METHOD: Kinematic data obtained from six healthy male subjects were used as inputs to a 'top-down' dynamic model. Each total segmental moment was partitioned into a net muscle component as well as gravitational and motion-dependent components. RESULTS: The model successfully predicted both muscle moments determined by the 'ground-up' inverse dynamics approach and also ground reaction forces measured with a force platform. Motion-dependent moments were consistently small compared with muscle and gravitational moments. CONCLUSION: Segmental motion during standing up is controlled almost entirely by gravitational moments and the direct action of muscles which span either end of the segment. Motion-dependent moments do not significantly reduce the muscle moments required to stand up. RELEVANCE: Knowledge and understanding of the mechanical demands on the body during rising from sitting to standing are important in identifying critical elements for successful performance. Such information may also provide a basis for the development or refinement of strategies to retrain the movement following injury or disease. PMID- 11415728 TI - Analysis of the sit-stand-sit movement cycle in normal subjects. AB - OBJECTIVE: This study aimed to establish a basis of descriptive data for the sit stand-sit movement cycle in 50 normal subjects, 25 male and 25 female, aged between 20.1 and 78.3 years (mean age 46.8 years). DESIGN: A descriptive design was employed to establish the characteristics of the activity in normal subjects. BACKGROUND: Research has been carried out into kinetic and kinematic characteristics of the sit-to-stand movement, but few researchers have considered stand-to-sit. Most studies have involved small samples, subjects with pathology, or elderly subjects, so a baseline of data from normal subjects has not yet been established. METHOD: Linear displacement and acceleration of the trunk and angular displacement of the knee were recorded simultaneously within the same temporal framework. The measurement system consisted of a vector stereograph, and triaxial accelerometers located at the level of C(7), and an electrogoniometer located at the lateral aspect of the knee. Subjects rose from and descended to the seated position a total of six times at their own self-selected speed. Numerical data were subjected to descriptive analysis, matched-pairs t tests and Pearson's rho correlations. RESULTS: Mean values for the time to rise was 1.91 s and to descend was 1.97 s. Forward lean velocity was greater during rising than descending (P < 0.001), and recovery velocity was greater during descending than rising (P < 0.001). Temporal contributions of forward lean and vertical displacement and the period of overlap between them were identified, and relationships between acceleration and temporal events and components were established. Differences existed among groups, involving primarily the elderly groups and occurring during the rising phase. CONCLUSIONS: This study has proposed a baseline of descriptive data in normal subjects for the sit-stand-sit movement cycle. RELEVANCE: In practical and clinical applications of information gained from analysis of functional activities, establishment of what is accepted as 'normal' is necessary before abnormalities can be identified and analysed, and intervention implemented and evaluated. This study provides that basis. PMID- 11415729 TI - Increased friction of animal joints by experimental degeneration and recovery by addition of hyaluronic acid. AB - OBJECTIVE: To investigate the lubricating ability in osteoarthritic (OA) stifle joints. DESIGN: An experimental study in rabbit stifles in vitro. BACKGROUND: The lubricating ability in OA joints, and the effects of the application of hyaluronic acid (HA), a drug for OA, to the osteoarthritic joints have not yet been reported. METHODS: The friction in OA stifles induced by two different methods, the intra-articular injection of papain, and transection of the anterior cruciate ligament (ACL) was measured using a robotic arm under force control. RESULTS: The frictional coefficient in the papain-injected model, that caused slight degeneration in the articular cartilage, was 1.3 to 1.8 times as large as that in the contralateral uninjected stifles. The elevated values decreased significantly by addition of HA to the articular surface (P < 0.05). In another group (n = 4), the frictional coefficient in the stifle at 9 months after ACL transection that caused severe degeneration in the articular cartilage was 1.6 to 7.8 times as large as that in the contralateral unoperated stifle. The addition of HA decreased the elevated values in three of the four ACL-transected joints. CONCLUSION: Friction was higher in the OA joints than in the healthy joints. The addition of HA was effective in the early stage of OA represented by papain injected case, while its effect seems not to be evident for advanced OA produced ACL transection which accompanies macroscopic shape changes. RELEVANCE: The present findings are relevant to the current research investigating the case of human osteoarthritis. We measured the frictional coefficients of experimentally degenerated rabbit stifles. The frictional coefficients can be used as an index of joint degeneration. We also assessed the recovery of the lubricating ability by addition of hyaluronic acid, a drug used for human osteoarthritis. PMID- 11415730 TI - Restriction of foot supination by ankle braces in sudden fall situations. AB - OBJECTIVE: To investigate the restriction of foot supination in different shoe orthotic combinations for unexpected ankle turns. DESIGN: A supination platform was used to experimentally induce sudden ankle turns. BACKGROUND: This study expanded on previous investigations of the effectiveness of different ankle braces and the influence of shoe material on foot supination. METHODS: For 21 male subjects pressure distribution data, achilles tendon angle, and supination velocity were collected. A comparison between a shoe with cut upper material and an intact shoe was done to show the influence of upper material on supination. The effectiveness of four different orthotics inside of an intact shoe were tested in comparison to the same shoe without ankle brace. Additionally, subjects rated their perceived supination movement of the foot. RESULTS: Between experimental conditions no large differences for the pressure distribution patterns were found. Three of the ankle braces reduced foot supination as well as supination velocities. More than two times lower supination values were measured for a footwear condition in which the upper material was cut away. Already during the free-fall most ankle braces caused a substantial reduction of foot inversion. CONCLUSION: The experiments demonstrated the influence of the upper material of a shoe and different orthotics on foot supination. Biomechanical measurements are necessary because of limited capabilities of subjects in detecting subtalar angular motions. RELEVANCE: This study investigated the effectiveness of different shoe conditions and ankle braces under experimental conditions that simulated unexpected ankle turns, the most frequent causes of sport injuries. The results of the study provide insights into which factors influence the amount of foot supination during unexpected ankle turns. PMID- 11415731 TI - The hybrid ring tubular external fixator: a biomechanical study. AB - OBJECTIVE: To measure and compare the mechanical properties in bending of the four-ring, and three-ring/one-tube hybrid external fixation frames. DESIGN. IN VITRO: measurements of the mechanical behaviour of ring and ring-tubular external fixation frames. In the latter, one ring of the full circular frame was replaced by one tube and Schanz screws. BACKGROUND: The mechanical properties of the classical Ilizarov four-ring external fixation frames has been compared to those of other external fixation frames by various authors. However, in clinical practice the hybrid fixation frame is being used with increasing frequency. Therefore the mechanical properties of the latter are of immediate interest and clinical value. METHODS: On explanted sheep tibiae with single and double osteotomies, frame stiffness in the four-point bending mode was measured at different K-wire tensions, comparing the values obtained from four-ring frames, to those of three-ring-tubular hybrid frames. These measurements were made under conditions of (a) bone distraction (BD), and (b) segment transport (ST), both at the initial and final stages of this procedure. RESULTS: In circular frames, frame stiffness in bending for increasing K-wire tension showed a Gaussian distribution both in distraction and post-ST with an optimum at 1000 N. In ring tubular hybrid frames, however, frame stiffness showed a more linear relationship to K-wire tension. CONCLUSIONS: In the four-ring Ilizarov external fixation frame, the exchange of one ring with one tube and one Schanz screw both reduced frame stiffness in bending and converted to linear its relationship to K-wire tension. RELEVANCE: Under clinical conditions, the use of a similar ring tubular hybrid external fixator allows the adjustment of frame stiffness in a simple and practical way. This is not the case with the original ring fixation frame. PMID- 11415732 TI - Patella position and biomechanical properties of the patellar tendon 1 year after removal of its central third. AB - OBJECTIVE: The aim of this study was to investigate how the removal of a patellar tendon graft for cruciate ligament reconstruction influences the mechanical properties of the remaining patellar tendon and the position of the patella at the knee joint. DESIGN: The experimental model of this in vivo study was the anterior cruciate ligament reconstruction in sheep. BACKGROUND: While the efficacy of a patellar tendon third as a ligament replacement has been extensively investigated, the consequences for the function of the remaining tendon and patella position remain unclear. METHODS: The central third of the patellar tendon from the right knees of 10 animals was removed for cruciate ligament replacement. One year postoperatively, the position of the patella within the joint and the stiffness and modulus of elasticity of the remaining tendon were compared with the non-operated contralateral controls. RESULTS: The patella position did not change in the operated knee joints in comparison with the controls. The cross-sectional area of the operated patellar tendon increased significantly by scar tissue formation at the defect site. CONCLUSION: Although the inferior mechanical properties of the scar tissue reduced the modulus of, elasticity of the tendon, the increase in cross-sectional area allowed it to develop a structural stiffness similar to the control tendons. RELEVANCE: The central third of the patellar tendon is often used for ligament replacement. Resulting biomechanical and structural changes in the remaining patellar tendon have not been previously reported using an in vivo animal model. The recovery of the biomechanical properties of the partially resected tendons allows for continued function. PMID- 11415733 TI - A Biomechanical Study on Five Unilateral External Fracture Fixation Devices. PMID- 11415734 TI - Biomechanics of the hip joint capsule -- a mathematical model and clinical implications. AB - OBJECTIVE: Our aim was to develop a mathematical model to calculate forces, tension and stretching in the hip joint capsule, under conditions caused by the joint effusion usually accompanying hip disease. DESIGN: A mathematical model was developed, based on experimental data from cadaver studies. BACKGROUND: Intracapsular pressure is important with respect to the degree of painless movement in the hip. Previously, we established the relations between the rotation around the axis of the neck of the femur, joint effusion, intracapsular pressure and joint stability. METHODS: In six cadaver adult hips the joint distraction, the traction force along and the rotation around the axis of the neck of the femur and the intracapsular pressure, were simultaneously monitored as the volume of intracapsularly infused saline was increased. The elasticity constants included in the extracted formula for the fluid pressure were calculated in a designed computer software based on these experimental data. RESULTS: Presented in Figures 3-12. CONCLUSIONS: In the normal joint there is no increase in intracapsular pressure, nor any tension in the hyperboloid shape capsule within the normal range of rotation around the axis of the neck of the femur. This shape is distorted in a hip with effusion, rotation then resulting in an increased intracapsular pressure and tension in the capsule, with potential risk of ruptures. PMID- 11415735 TI - Assessment of bone quantity and 'quality' by ultrasound attenuation and velocity in the heel. AB - OBJECTIVE: To determine if ultrasound measurements in the heel are related to bone quality in addition to quantity. DESIGN: In situ and in vitro experiments on cadaver heels. BACKGROUND: It has been suggested, but not demonstrated, that clinical ultrasound - used to screen for osteoporosis in clinical trials - provides a measure of 'bone quality' as distinct from bone quantity. METHODS: Ultrasound transmission velocity (UTV) and the slope of the linear dependence of broadband ultrasound attenuation on frequency (BUA) were measured in situ in 32 heels of 16 cadavers and in vitro in cores of calcaneal trabecular bone. RESULTS: After adjusting for Young's modulus, in situ UTV explains 33% (P = 0.03) and in situ BUA explains none of the remaining variance in density (r(2) = 0.02, P = 0.60). After adjusting for density, in situ BUA explains 29% (P = 0.04) and in situ UTV explains none of the remaining variance in Young's modulus (r(2) = 0.01, P = 0.79). By comparison, in vitro BUA explains 58% (P = 0.001) of the remaining variance in Young's modulus, after adjusting for density. CONCLUSIONS: In situ BUA reflects 'bone quality' independently of bone quantity, whereas in situ UTV reflects bone quantity independently of 'bone quality'. PMID- 11415736 TI - Valgus-varus motion of the knee in normal level walking and stair climbing. AB - OBJECTIVE: The knee valgus-varus moment and the knee angles were compared between normal level walking and stair climbing. DESIGN: Ten healthy subjects were tested for ascent, descent, and level walking. BACKGROUND: An understanding of the normal valgus-varus motion of the knee during stair climbing is needed to apply biomechanical analysis of stair climbing as a evaluation tool for knee osteoarthritis patients. METHODS: A motion analysis system, three force plates, and a flight of stairs were used to collect kinematic and kinetic data. The knee angles and moments were calculated from the collected kinematic and kinetic data. RESULTS: The knee varus angle for the maximum knee valgus moments in stair climbing was significantly greater than that in level walking. The knee valgus moment was significantly correlated to ground reaction forces and knee valgus varus angle during stair climbing and level walking. CONCLUSIONS: There is a coupling between the knee valgus-varus motion and flexion-extension motion. Ground reaction forces are the major contributors to the within-subject variation in the knee valgus-varus moment during stair climbing and level walking. The knee valgus-varus angle is a major contributor to the between-subject variation in the knee valgus moment during stair climbing and level walking. PMID- 11415737 TI - Perceptual and biomechanical variables for running in identical shoe constructions with varying midsole hardness. AB - OBJECTIVE: Perceptual ratings of mechanical variables were compared with biomechanical variables that are related to running injuries. DESIGN: Eight identical running shoes with a relatively close range of midsole hardness were used. Ground reaction force (GRF), in-shoe pressure distribution and rearfoot motion were measured during running. Perceptual ratings were obtained after the running trials. BACKGROUND: Previous studies reported high correlations between cushioning perception and biomechanical variables for shoes that featured large differences in midsole hardness. METHODS: A 15-point categorical rating scale was used to judge impact severity, pressure magnitude and rearfoot motion in running. Rating scores were compared with biomechanical variables (GRF, pressure distribution and pronation values) using regression analyses. RESULTS: Regression analyses revealed high relations between different biomechanical variables and the perception scores. The best relation to perception was analysed for the median power frequency of the vertical GRF (r(2) = 0.97). A negative correlation (r(2) = 0.54) between the first impact of GRF and the perception of impact severity could be revealed. CONCLUSION: The present study suggests that the body's sensory system seems to differentiate well between impacts of different frequency content. Based on perceptual abilities, subjects adapt their running style to avoid high heel impacts. PMID- 11415738 TI - Dynamic loading affects the mechanical properties and failure site of porcine spines. AB - OBJECTIVE: The purpose of this study was to investigate the effect of load rate on the mechanical characteristics of spinal motion segments under compressive loading. DESIGN: An in vitro experiment using a porcine model which ensured a homogeneous population for age, weight, genetic background and physical activity. BACKGROUND: Spinal motion segments comprise of viscoelastic materials, and as a result the rate of loading will modulate mechanical characteristics and fracture patterns of the segments. METHODS: Twenty-six cervical porcine spines were excised immediately post-mortem with all soft tissue intact. Spines were then separated into two specimens each consisting of three vertebral bodies and the two intervening intervertebral discs (C2-C4 and C5-C7) and loaded to failure under five loading rates (100, 1000, 3000, 10 000 and 16 000 N s(-1)). After the specimens failed, they were dissected to determine the mode of failure. RESULTS: Dynamic loading increases the ultimate load compared with quasi-static loading (100 N s(-1)), whereas the magnitude of dynamic loading (1000-16 000 N s(-1)) appears not to have a significant affect. Stiffness behaved in a similar manner. The displacement to failure of specimens decreased as load rate increased, although there was a diminishing effect at high load rates. Furthermore, failure at low load rates occurred exclusively in the endplate, whereas failure of the vertebral body appeared with greater frequency at higher load rates. CONCLUSIONS: The mechanical characteristics and resulting injuries of porcine spinal motion segments were affected as the loading rates changed from quasi-static to dynamic. The modulating factors of the mechanical characteristics of the spine need to be understood if valid models are to be designed which will increase the understanding of spinal function, and are important for choosing better injury prevention and rehabilitation programmes. PMID- 11415739 TI - Fatigue-related EMG responses of trunk muscles to a prolonged, isometric twist exertion. AB - OBJECTIVE: To determine the responses of the trunk muscles to fatigue during sustained, isometric axial torque exertions. DESIGN: Electromyographic (EMG) amplitude and frequency parameters were used to investigate the effects of prolonged contractions on the recruitment patterns of trunk muscles, with special emphasis on those antagonistic and stabilizing muscles not contributing directly to the required axial torque. BACKGROUND: High levels of muscle coactivity have been observed during axial torque generation in the trunk. It has been suggested that this serves to enhance postural stability of the spine during potentially injurious twist exertions. Muscle fatigue decreases force-generating capacity but the resulting effects on muscle recruitment, and the subsequent consequences for postural stability, have not been explored previously during the production of axial torques in the spine. METHODS: Eleven male and 11 female subjects maintained an isometric axial torque exertion to the left (40% of maximum) until volitional exhaustion in an upright standing posture. Maximum axial torques were measured before and after the trial. The average EMG amplitude (AEMG) and mean power frequency (MPF) of seven bilateral trunk muscles (representing agonists, antagonists and trunk stabilizers) were continuously monitored throughout the trial. RESULTS: The average decrease in maximum torque was 18.5% and the average endurance time was 163 +/- 52 s. Evidence of muscle fatigue was provided by significant MPF decreases in 12 of the 14 muscles monitored (P < 0.05) with a overall average decrease of 20.4%. There was a significant increase in AEMG with time for 11 of the 14 muscles monitored (P < 0.05). These increases in activation were linked to increases in muscle force for most of the antagonistic and stabilizing muscles. Gender effects were rarely observed. CONCLUSIONS: Fatigue results in an change in the recruitment patterns of trunk muscles. Muscles serving as antagonists and trunk stabilizers during prolonged axial twist exertions increased their force levels in response to fatigue. PMID- 11415740 TI - Development and application of predictive equations of maximal static moments generated by the trunk musculature. AB - OBJECTIVE: To develop and apply regression equations that predict the maximal static moments generated by the extensor, axial rotator and lateral flexor muscles of the trunk as a function of forward bending angles and asymmetrical positions of the torso in human subjects. DESIGN: Using experimental data, polynomial predictive equations were developed. The equations were applied in an algorithm to estimate the maximal moment generated by individual subjects. METHODS: Three groups of male subjects participated in the evaluation of the effect of forward bending and trunk asymmetry on the static moment of the extensor, right axial rotator, and right lateral flexor muscles of the trunk, respectively. Two other groups of male subjects were evaluated to test the validity of the algorithm. During the dynamometric tests, the three-dimensional position of the trunk was determined using cinematography techniques. RESULTS: For the extensor muscles, a curvilinear increase of the maximal moment was observed with forward bending, with a slight decrease of the moment on each side of the neutral position. For the right axial rotator muscles, both forward bending and left asymmetrical position of the trunk increased the maximal moments generated by these muscles. The right lateral flexors were stronger in the standing and mid-flexion positions than in other positions. First-order, second order and interaction terms appear in the regression equations to account for the effect of forward bending and asymmetrical positions. The regression equation for the extensors and right axial rotator muscles have R(2) coefficients (range 0.56 0.76) higher than those of the lateral flexor regression equations (range 0.09 0.12). The algorithm results in an average error of 13% in predicting the strength of extensor and axial rotator muscles of individual subjects. CONCLUSION: The results have shown that strength of the extensor and axial rotator muscles is affected by forward bending and the asymmetry of the trunk, while strength of the lateral flexor muscles is only dependent on the forward bending position. The regression equations and the corresponding algorithm are applicable for the range of positions used in the study. PMID- 11415741 TI - Lumbar mobility in former elite male weight-lifters, soccer players, long distance runners and shooters. AB - OBJECTIVE: To investigate the long-term effects of different loading conditions in sports and work on lumbar mobility. DESIGN: Factors associated with lumbar mobility were sought by examining sports experience, occupational history, back pain history, anthropometric characteristics, and lumbar magnetic resonance images in 114 former Finnish male elite athletes: 30 soccer players, 29 weight lifters, 27 long-distance runners, and 28 shooters, aged 45-68 yr. BACKGROUND: Back pain and aging reduce spinal mobility, whereas some sports may increase it. Little is known about the effects of common loading conditions found in work and sports, that do not require extremes of spinal motion on lumbar mobility later in life. METHODS: Sports and occupational histories were obtained from a standardized interview. Lumbar mobility was measured by the flexicurve method. Disc degeneration was evaluated from lumbar magnetic resonance images. The results were analysed with analysis of variance and covariance and multiple regression models. RESULTS: The athlete groups did not differ significantly in lumbar sagittal mobility. Higher body-mass index was associated with less flexion, reduced disc height with less extension, as was a high lifetime number of low back pain episodes. Occupations characterized by varying work postures and light lifting were associated with greater mobility, and heavy work with lesser mobility. CONCLUSIONS: Participation in sports with clearly different loading patterns, that do not emphasize extremes of spinal range of motion, do not appear to lead to significant differences in back mobility in later adulthood, but occupational loading factors and disc height narrowing appear to influence spinal mobility. PMID- 11415742 TI - Distribution and regional strength of trabecular bone in the porcine lumbar spine. AB - OBJECTIVE: The regional strength and distribution of trabecular bone in the porcine lumbar spine were examined to understand the mechanical responses in quadrupedal vertebrae. DESIGN: The cancellous bone columns prepared from different regions of porcine lumbar vertebrae were subjected to axial compression to investigate the regional strength related to disc structure. Some vertebrae were also serially sectioned sagittally and transversely to observe the trabecular patterns. BACKGROUND: Animal spines were often used in biomechanical testing and the results were said to be similar to those of humans. However, none of them are truly like human bipedal locomotion. The understanding of regional variations in compressive properties may help us to explain the similarity and interpret the data of animal experiments. METHODS: The dried, defatted cancellous bone columns were subjected to uniaxial compression of different regions at a deformation rate of 5 mm/min. The mechanical properties in different regions were compared. The main trabecular orientation was also observed using a zoom stereo microscope. RESULTS: The ultimate strength of the posterior column tended to be larger than that of the anterior column. The ratio of bony strength overlying the nucleus pulposus to that overlying the annulus fibrosis was constant and averaged 1.16. The main trabecular struts were noted to be parallel to the spinal canal and cross-bridged by thinner trabeculae. CONCLUSIONS: The trabecular morphology and regional mechanical properties of the porcine spine were comparable to those of other similar studies on the human spine. Its significance needed further study. PMID- 11415743 TI - Measurements of vertebral translations using bone pins, surface markers and accelerometers. AB - OBJECTIVE: To compare invasive and non-invasive techniques for measuring the posterior-to-anterior translations of vertebrae during spinal manipulative therapy. DESIGN: This study represents a small part of a larger experiment. BACKGROUND: Despite the mechanical nature of spinal manipulative therapy, the mechanism by which it alleviates back pain is still unknown. An understanding of the deformation behaviour of the spine during spinal manipulative therapy would aid in the formulation of a hypothesis underlying its efficacy. METHODS: A clinician delivered posterior-to-anterior manipulative thrusts to the right transverse process of either T(10), T(11) or T(12) in two cadavers. Posterior-to anterior translations of the bone pins and surface markers (embedded in and taped over T(10), T(11) and T(12), respectively) were recorded by cine cameras. Posterior-to-anterior accelerations, recorded by the accelerometers, were used to calculate posterior-to-anterior translations. Translation measurements made by using the surface markers and the accelerometers were compared to those made by using the bone pins. RESULTS: There were no significant differences between the posterior-to-anterior translations of vertebrae obtained from the surface markers, as compared to the bone pins. The accelerometers underestimated the absolute, and overestimated the relative, vertebral translations, respectively, compared to the bone pins. CONCLUSIONS: The translations measured by the surface markers were more similar to the translations derived from the bone pins than those calculated from the accelerometers. Three-dimensional surface marker arrays would therefore be more useful to determine all relative movements non invasively. PMID- 11415744 TI - Finite element models in tissue mechanics and orthopaedic implant design. AB - This article attempts to review the literature on finite element modelling in three areas of biomechanics: (i) analysis of the skeleton, (ii) analysis and design of orthopaedic devices and (iii) analysis of tissue growth, remodelling and degeneration. It is shown that the method applied to bone and soft tissue has allowed researchers to predict the deformations of musculoskeletal structures and to explore biophysical stimuli within tissues at the cellular level. Next, the contribution of finite element modelling to the scientific understanding of joint replacement is reviewed. Finally, it is shown that, by incorporating finite element models into iterative computer procedures, adaptive biological processes can be simulated opening an exciting field of research by allowing scientists to test proposed 'rules' or 'algorithms' for tissue growth, adaptation and degeneration. These algorithms have been used to explore the mechanical basis of processes such as bone remodelling, fracture healing and osteoporosis. RELEVANCE: With faster computers and more reliable software, computer simulation is becoming an important tool of orthopaedic research. Future research programmes will use computer simulation to reduce the reliance on animal experimentation, and to complement clinical trials. PMID- 11415745 TI - Two simple methods for improving the reliability of joint center locations. AB - OBJECTIVE: A clinically oriented technique is proposed for evaluating the reliability of methods for estimating joint center locations from surface markers, as is an optimization method for estimating joint center locations during planar movements. DESIGN: Segment length variability is used as a measure of reliability, and three simple methods for locating joint centers are compared via repeated measures analysis. Rigorous evaluation is achieved by applying adjustment parameters to a data set, other than the one from which parameters were derived. BACKGROUND: Although more sophisticated techniques are available, many clinical and experimental studies use visual observation and palpation to locate joint centers. This study offers a simple means to evaluate the reliability of that method, and it offers two simple post-hoc methods to improve reliability. METHODS: Single-joint movements are used to generate adjustment parameters from three-dimensional (3D) measurements of surface markers; these are applied to multi-joint movement trials. Segment length variability is compared before and after adjustment with each of two post-hoc methods. RESULTS: As shown by lowered segment length standard deviations, the proposed optimization technique improved reliability compared to the observational and the two dimensional (2D) post-hoc methods. CONCLUSIONS: The segment length technique offers a simple means to evaluate the reliability with which joint centers are located, and the new optimization method improves reliability for planar multi joint movements. RELEVANCE: These improvements in reliability are easily implemented within settings where sophisticated technical support may be unavailable. PMID- 11415746 TI - Changes in knee joint function over a wide range of walking speeds. AB - OBJECTIVE: Changes in patterns of knee joint flexion-extension rotations, moments and powers were investigated during the stance phase across a wide range of walking speeds that includes the very slow speeds used by many patients referred for gait analysis. DESIGN: Each subject walked at 25, 50, 75, 100, and 125% of a scaled natural speed of 0.785 statures.s(-1). BACKGROUND: The results of earlier studies suggest that control subjects, as they walk at progressively slower speeds, increasingly use a large internal knee flexor moment in the early stance phase. METHODS: Eighteen healthy adult subjects (nine male and nine female) were tested. RESULTS: The shapes of the mean patterns were similar for the three fastest, most natural walking speeds. At the two slowest speeds, however, subjects used little knee flexion and small knee moments through mid-stance, resulting in negligible joint power through the first 80% of the stance phase. At these speeds, most subjects had moment patterns that could not be classified with confidence as 'normal' or as predominantly flexor or extensor. Only four subjects had knee flexor moment patterns at the slowest speeds, and these included much smaller flexor moments in the early stance phase than in some previous reports. RELEVANCE: Many patients walk at very slow speeds for which 'normal' joint kinetic patterns have not been described. The knee joint kinetic data provided here from healthy subjects is useful in helping to distinguish 'abnormal' patterns caused by neuromuscular pathologies or adaptations from patterns reflecting the effects of slow walking speed alone. PMID- 11415747 TI - The relationship between elbow flexor volume and angular stiffness at the elbow. AB - OBJECTIVE: The purpose of this experiment was to determine if a correlation exists between the volume of the elbow flexors and angular stiffness at the elbow, and to determine the contribution of the biceps brachii and the brachialis muscles to angular stiffness. DESIGN: This study is a descriptive, correlational study and presents a graphical model of the passive properties of muscle. BACKGROUND: The correlation between arm volume and angular stiffness has been shown, but the measurement of arm volume was not specific to the structures being strained. Pre-positioning a bi-articular muscle by stretching over one joint decreases the range of motion at the other joint and may affect the stiffness. METHODS: Angular stiffness at the elbow of 14 female and 15 male volunteers was measured, and the volume of the elbow flexors was calculated from compounded ultrasound imaging. Initial biceps length was set by pre-positioning the shoulder in two different positions. RESULTS: A significant linear relationship was observed between the slope of phase 1 of the stiffness curve and volume of the elbow flexors in both horizontal flexion (r = 0.92) and horizontal extension (r = 0.79) of the shoulder. Phase 2 of the stiffness curve showed no linear relationship to muscle volume in either shoulder position (flexion, r = 0.22; extension r = 0.33). The slopes of phases 1 and 2 were significantly greater with the shoulder in horizontal extension than in horizontal flexion. CONCLUSION: The volume of the elbow flexor muscles is a good predictor of angular stiffness in phase 1 of the curve. A model of the additive contribution of the biceps and brachialis muscles is presented to account for the increased stiffness in the shoulder extended position. RELEVANCE: Recognition of the correlation between muscle volume and stiffness, coupled with understanding that pre-positioning a bi articular muscle may affect muscle stiffness may aid the clinician in accurately assessing muscle stiffness in patients with connective tissue disorders, neurological dysfunction and contractures. PMID- 11415748 TI - Variability of forces applied by experienced therapists during spinal mobilization. AB - OBJECTIVE: To determine the variation in forces used by different therapists during mobilization of the lumbar spine and the repeatability and reproducibility of individual therapists. DESIGN: An instrumented mobilization couch was developed to measure the forces applied to the trunk during spinal mobilization. BACKGROUND: Due to limitations in equipment design and data analysis, previous related studies demonstrate equivocal results. METHODS: The system was used to collect data from a sample of 30 experienced therapists to evaluate variation, repeatability and reproducibility during the application of five mobilization procedures. RESULTS: The variation in forces used by different therapists when performing the same technique was substantial, ranging between 63 and 347 N for one technique. During this procedure, 30% of the therapists were found to be relatively consistent, repeating the magnitude of the force applied at the first session within 5%. Others demonstrated considerable variation, exhibiting a difference as great as 34%. CONCLUSIONS: The inconsistency between experienced therapists has considerable implications for clinical practice. Changes in the magnitude and rate of loading are likely to have different effects due to the inherent viscoelastic behaviour of soft tissues. RELEVANCE: Spinal mobilization and manipulation techniques are frequently used by manual therapists in the treatment of musculoskeletal disorders. Despite the reliance on these techniques in clinical practice, there is little scientific evidence to substantiate their use. Before progress in this area can be made, it is necessary to characterize the forces used during typical mobilization procedures. The results can be used to develop teaching strategies and as a basis for comparative research on the efficacy of these techniques. PMID- 11415749 TI - An in vivo study of the intervertebral movements produced by posteroanterior mobilization. AB - OBJECTIVE: To measure the intervertebral movements of the lumbosacral spine when it was subjected to posteroanterior mobilization. DESIGN: An in vivo radiographic study. BACKGROUND: Posteroanterior mobilization is frequently employed in the examination and treatment of low back pain, but its mechanical and therapeutic effects are uncertain. METHODS: Lateral radiographs were taken of 12 healthy male subjects who were in prone lying. This was repeated while a static mobilization force of 150 N was applied vertically to their L4 spinous processes. The radiographic images of the vertebrae were traced and digitized. The sagittal rotation and translations of the motion segments, and the displacements of the spinous processes were then computed. RESULTS: Under the application of the mobilization loads, the lumbar motion segments were found to extend, whereas the L5/S1 segment tended to flex. It was also shown that the upper lumbar segments (L1/2-L3/4) translated posteriorly and the L5/S1 segment anteriorly. The L4/5 segment did not exhibit translation in a consistent direction. In addition, all the spinous processes were found to be displaced anteriorly. CONCLUSIONS: The movement pattern observed in this study strongly suggests that the spine is subjected to three-point bending under the application of mobilization loads. RELEVANCE: The intervertebral movements produced by posteroanterior mobilization were small in magnitude, and it is unlikely that the therapist can assess these movements manually with reasonable reliability. Mobilization should be interpreted as a passive test of the whole lumbar spine in three-point bending. PMID- 11415750 TI - Spinal shrinkage during work in a sitting posture compared to work in a standing posture. AB - OBJECTIVE: The objective was to measure the possible differences in shrinkage of the thoracolumbar spine in subjects working in a sitting and a standing posture for 6.5 h at work, in a realistic work environment. The isolated shrinkage of the thoracic and the lumbar spine was also examined. STUDY DESIGN: This study presents a new protocol to measure shrinkage of the thoracic and lumbar spine separately. BACKGROUND: Controversies still exist with regard to the load on the spine in a sitting compared to a standing position. Some report that shrinkage is greatest in the sitting position while others report the reverse. However, nothing is known about the height reduction of the thoracic and the lumbar spine during loading for 6.5 h in a real work environment. Therefore, the behaviour of the thoracic and the lumbar spine under practical condition has to be investigated. METHOD: A stadiometer with a measurement error of 0.51 mm was used to measure changes in spinal height during work. To exclude first-time behaviour of the spine, a pre-test lasting 50 min was undertaken. The mean of the last three measurements was used as the reference height. During work, height measurements of the spine were performed every 20 min. To separate the behaviour of the thoracic and the lumbar spine, two benchmarks were placed at the vertebrae prominens and at the thoracic-lumbar junction. Shrinkage of the spine was investigated within three different cohorts: (I) work in a sitting posture for 6.5 h; (II) relaxed sitting for 2 h vs work for 2 h in a sitting position and (III) work in a standing position for 6.5 h. RESULTS: Relaxed sitting leads to a gain in stature compared to work in a sitting position for 2 h. The major gain in stature occurred in the lumbar spine. Comparison of cohort (III) working in a standing position with cohort (I) working in a sitting position shows that the shrinkage of the spine is greatest when work is performed in a standing posture. The major differences were found in the shrinkage of the lumbar spine, e.g. shrinkage of the lumbar spine in the standing cohort (III) was 4.16 mm compared to 1.73 mm in the sitting cohort (I). CONCLUSIONS: There is a gain in stature during relaxed sitting compared to work in a sitting posture. The load on the spine is greatest when work in a standing position is performed. The greater shrinkage of the lumbar spine during work in a standing position compared to a sitting posture is probably due to: (i) differences in lumbar lordosis and (ii) the effect of bending and torsion while handling the work materials. PMID- 11415751 TI - Box tilt and knee motions in manual lifting: two differential factors in expert and novice workers. AB - OBJECTIVE: It was the objective of this study to investigate the kinematic and kinetic effects of two specific handling factors that differentiate expert and novice workers, namely the level of knee flexion and box tilt. DESIGN: Seven inexperienced subjects were required to lift a 12-kg box in the sagittal plane using three different strategies: (1) reduced knee flexion and a backward box tilt (more typical of experts); (2) large knee flexion and a backward box tilt; (3) large knee flexion and no box tilt (more typical of novices). BACKGROUND: The lifting techniques of highly skilled workers differ substantially from those of novices but only limited information is available to compare their biomechanical differences. METHODS: The methods included dynamic segmental analyses to calculate the net moments at all body joints and a planar single-muscle equivalent to estimate compression loadings at L5/S1; total work and joint work distribution were calculated using the integration of joint power. The 3-D kinematic data were acquired with three video cameras and force data were obtained with one AMTI force platform which were synchronized with the film data. RESULTS: Box tilt reduced the load trajectory and loadings on the lower back and shoulders; a reduced knee flexion affected body posture and reduced mechanical work and loadings on all body joints. The combination of these two factors, box tilt and reduced knee flexion, showed added effects for almost all variables. CONCLUSIONS: These two factors proper to experts were, in this context, biomechanically more advantageous. PMID- 11415752 TI - Postural sway and stepping response among working population: reproducibility, long-term stability, and associations with symptoms of the low back. AB - OBJECTIVE: To investigate the day-to-day reproducibility and long-term (9 months) stability of variables of postural control, and the associations of these variables with low-back and lower-extremity pain in a working population. DESIGN: Test-retest measurements of 18 healthy subjects. Cross-sectional study of 165 working women and 343 men. BACKGROUND: Sudden loss of postural balance may result in microtraumas of back structures. Therefore workers with decreased postural control may run an increased risk of low-back pain. There are few reports describing the reproducibility of force-plate-based posturography. METHODS: Amplitude and velocity of postural sway were measured with a force-plate in a two feet stance with eyes open and closed, and in a one-foot stance with eyes open. A stepping response test was developed to measure the dynamic components of balance. RESULTS: The mean differences between the repeated measurements were generally 5-10% and the standard deviations of these differences were up to a quarter or one third of the mean measurement values. Sway velocity showed the best overall reproducibility. Men had a larger sway than women independent of age and anthropometry. The group of non-symptomatic subjects showed wide variation and a slightly lower sway than the groups with low-back or lower-extremity pain. CONCLUSIONS: Postural sway has moderate stability, wide variation, and slight associations with low-back symptoms in a working population. PMID- 11415753 TI - Annular bulge contours from an axial photogrammetric method. AB - OBJECTIVE: To determine the characteristic contour of annular bulge in the lumbar spine under compression using a non-contacting technique. DESIGN: Transverse bulge of the annulus in lumbar motion segments was measured from photographs taken through an acrylic load plate. BACKGROUND: Annular bulge is considered a potential factor in nerve root impingement. However, consensus is lacking regarding which annular region bulges most. Previous studies have relied on markers in discrete locations or on contacting techniques, both of which limit the accuracy of bulge measurements. Reporting of continuous contours around the periphery of the disc has been limited to individual cases; characteristic behavior derived from a statistical treatment of the data is unknown. METHODS: Sixteen lumbar discal motion segments were loaded to 2500 N in compression. Axial photographs were taken in the pre-loaded and loaded states and digitized with an effective accuracy of 0.05 mm at 5 degrees intervals. The bulge was averaged over the sample of specimens at each interval to provide a mean-value plot in the transverse plane. Regional variations were evaluated statistically. RESULTS: It was found that the bulge was greatest posterolaterally (0.93 mm, SD 0.52 mm), and smallest anteriorly (0.37 mm, SD 0.26 mm; corrected p < 0.05), with an average of all data points of 0.67 mm, SD 0.42 mm. The restraining influence of the posterior longitudinal ligament, while not statistically significant, was evident in magnified view of the bulge contour. These regional differences were greater in the lower than in the upper lumbar segments. CONCLUSIONS: Herniation posterolaterally may be associated with relatively large annular bulge in that region, although normal bulge is only of the order of 1/100th the diameter of the disc. PMID- 11415754 TI - Differences in mechanical response between fractured and non-fractured spines under high-speed impact. AB - OBJECTIVE: The differences in mechanical response between fractured and non fractured spines were investigated using a porcine spine impact model. DESIGN: Ten three-vertebrae segments (C3-C5) of porcine spine were subjected to a single impact to study the trauma mechanism. Small steel balls glued to the vertebra and a high-speed camera were used to observe the deformation of vertebral body and disc during impact. After trauma, the episodes of fractured specimens were compared with those of non-fractured specimens. BACKGROUND: Experimental trauma models using the spines of mature animals have rarely been evaluated. Finding a well-controlled, reproducible protocol based on an easily accessible specimen was therefore important. These models will be promising if clinical fractures can be produced. METHODS: All of the specimens were subjected to high-speed flexion compression loading. The impact to the load cell and the operation of the high speed camera were synchronized. The force-time sequence and disc deformation curve were recorded. The results from fractured and non-fractured spines were then compared. RESULTS: There were three burst fractures, four pedicle fractures, one facet joint fracture, one compression fracture and one fracture-dislocation. All of these fractures were similar to clinical fractures. Compared to non fractured specimens, the fractured specimens had lower maximal force and longer reaction time. The characteristic steep decline in the middle region of the force time curve was also consistently noted in the fractured spines. CONCLUSIONS: Spinal fractures similar to those found clinically were successfully produced in porcine spines. The characteristics of the mechanical responses observed should be helpful in the interpretation of events which occur during impact. PMID- 11415755 TI - A method to determine joint moments and force distributions in the shoulders during ceiling work -- a study on house painters. AB - OBJECTIVE: The purpose of this study was to propose and apply a method to study the net load and the force distribution in the shoulder during sanding of a ceiling. DESIGN: Sanding of a ceiling was studied in an experimental set-up. BACKGROUND: Among painters there is a high frequency of neck and shoulder complaints and some particularly strenuous tasks are sanding and painting of ceilings with tools attached to extension handles. METHODS: A biomechanical model predicted the load pattern in the shoulder from measured external forces and body postures. Measurements were made on 40 subjects by means of a force plate, an extension handle supplied with two pairs of strain gauges and a load cell, and a 3D optoelectronic measurement system. RESULTS: Work techniques may seem to be similar at visual inspection, but nevertheless produce quite different loading patterns in the shoulder. The observed work techniques varied largely among the subjects but the intra-individual variability was rather small. CONCLUSIONS: The study showed that the proposed method can be used to evaluate loads and force distribution in the shoulder. Since the method involves the use of a biomechanical model that can scarcely be validated, results must be interpreted cautiously. PMID- 11415756 TI - The variability of shoulder motions recorded by means of palpation. AB - OBJECTIVE: The objective of this methodological study is the quantification of the sources of variability in the recorded three-dimensional motions of the shoulder mechanism for comparative purposes. BACKGROUND: The palpation and subsequent digitization of skeletal landmarks of the shoulder mechanism is a non invasive and relatively easy method to quantify shoulder orientations. Comparison of individual motions is subject to the accuracy of the palpation method, the magnitude of kinematic redundancy of the shoulder mechanism and inter-subject differences in morphology and physiology. Quantification of these sources of variance, i.e. the palpation error, motoric noise and inter-subject differences, demonstrated the accuracy of the method and the potential validity of the descriptive motion parameters, e.g. Cardan angles, in intra- and inter-individual studies for clinical, ergonomical and biomechanical studies. METHODS: The orientations of the shoulder bones were recorded five times for each of five subjects by palpation and digitization of 12 skeletal landmarks for 10 equidistant arm elevation postures in the scapular plane. The orientations were described by means of Cardan angles. The palpation error was determined at a standardized initial rest position and expressed by Cardan angles for each recorded posture. Adding motoric noise and inter-subject differences gave the inter-individual variance. RESULTS: The palpation error was approximately 2 degrees. The major recorded variance originated from motoric noise (+/-33%) and inter-subject difference (+/-55%). CONCLUSIONS: The palpation method is an accurate means of recording the three-dimensional orientations of the shoulder mechanism and for intra-individual studies. However, inter-subject variability is large. PMID- 11415757 TI - The measurement of isokinetic fingers flexion strength. AB - OBJECTIVE: To investigate the feasibility of using isokinetic dynamometry for measuring the dynamic strength of the fingers flexors. SUBJECTS: Sixteen healthy women aged 19-52. DESIGN: Flexion strength was measured isometrically using a Jamar dynamometer and isokinetically using a special attachment which was harnessed to a KinCom 125 dynamometer. In the series comprising the dynamic measurements, the elements of contraction mode (concentric vs eccentric), test velocity (low, medium and high) and isometric preactivation bias (low vs high) were incorporated. RESULTS: Dynamic strength parameters manifested expected physiological relationships both within their own frame of reference and with respect to the static measures. CONCLUSION: Based on various facets of validity it is suggested that isokinetic dynamometry offers a valid venue for measuring fingers flexion strength. Furthermore, this study allows a selection of test parameters for the standardization of this measurement. PMID- 11415758 TI - New concepts -- biomechanical studies of a newly designed femoral prosthesis (cervico-trochanter prosthesis). AB - OBJECTIVE: A newly designed stemless (cervico-trochanter) prosthesis was developed for the purpose of reducing the incidence of the stress-shielding effect caused by the traditional stem-type prosthesis. DESIGN: Both mechanical test and three-dimensional finite element analysis were performed for comparing the differences of strain and stress distributions between the intact, C-T and PCA prosthetic femora. BACKGROUND: The stress-shielding effect and polyethylene (PE) wear debris were thought to be the main factors that resulted in local bone loss after the implantation of stem-type prostheses. In this study, we developed the new C-T prosthesis, which aimed to resolve the above-mentioned problems. METHODS: Six pairs of femora taken from human male cadavers were used to compare the strain magnitudes of intact (n = 12), C-T (n= 12) and PCA (n = 4) prosthetic femora in specific positions. Failure load tests of C-T (n = 8) and PCA (n = 12) prosthetic femora were also carried out from the load-displacement curve. The analysis of variance (ANOVA) test was used for statistical analysis. In addition, three-dimensional finite element stress analyses were performed using a commercial package, ANSYS, on a Convex 3810 computer. RESULTS: Both mechanical test and finite element results showed that the C-T prosthetic femora has a lower stress-shielding tendency than the PCA prosthetic femora (P < 0.001). The C-T prosthetic femora also withstood an average bearing load of 6312 N, which is greater than that of the PCA prosthesis at 5358 N (P < 0.01). CONCLUSIONS: The C T prosthetic femur could withstand a higher failure load than the PCA prosthesis, which effectively reduced the incidence of the stress-shielding effect. Moreover, the particular design of the C-T prosthesis also reduced localized osteolysis because of the overall coverage of the neck-trochanteric area. PMID- 11415759 TI - Comparison of hip center estimation using in-vivo and ex-vivo measurements from the same subject. AB - OBJECTIVE: To evaluate the accuracy of locating the hip center (HIPC) by comparing in-vivo estimations to posthumous HIPC measurement in the same subject. BACKGROUND: Numerous techniques have been used to locate the HIPC in living subjects and in cadavers. There are no published reports, however, which have compared measurements of the HIPC obtained in-vivo and ex-vivo for the same subject. DESIGN AND METHODS: Bilateral hip centers (HIPC) were estimated for an elderly male patient seven times over 32 months in a gait laboratory using a technique which combines kinematic data and static pointing. The patient's pelvis was recovered posthumously and analytical photogrammetry used to estimate the HIPC locations. Published methods which scale anatomical measurements to locate the HIPC were compared to our ex-vivo and in-vivo measurements. RESULTS: HIPC precision was 0.94 cm for the in-vivo measurements (N = 7 trial days) and 0.15 cm for the ex-vivo measurements (N = 6 photogrammetry trials). The resultant error between ex-vivo and in-vivo estimates of HIPC position was 1.17 cm. Resultant errors in locating the HIPC using published anatomical scaling factors ranged between 1.48 cm and 5.37 cm. CONCLUSIONS: Although we have been unable to compare in-vivo and ex-vivo HIPC measurements for more than one subject, results of our analysis suggest that using anatomical scaling to locate the HIPC may result in unsatisfactory subject-specific estimates of the HIPC. PMID- 11415760 TI - Contact patterns at the tarsal joints. AB - OBJECTIVE: To determine the contact patterns at the tarsal joints under two levels of loading (150 and 600 N) with the foot in the neutral position, plantarflexion, dorsiflexion, inversion and eversion. DESIGN: In vitro measurements quantifying the size, location and patterns of contact. BACKGROUND: Several methods have been employed to study contact areas and patterns in joints such as the hip and knee; in contrast there is limited data on the joints of the foot. METHODS: Contact areas and patterns were determined by injecting coloured dyes, one for each level of loading, into the joint spaces in 23 pairs of feet: three pairs in the neutral position, five pairs in each of plantarflexion and dorsiflexion, and ten pairs in inversion (right foot) and eversion (left foot). RESULTS: Contact area maps were constructed for the ankle, subtalar, talonavicular, calcaneocuboid, cuneonavicular and the intercuneiform joints. Quantification of the surface areas was performed using a digitizer. The extent of the changes in contact area appeared to be related to the mobility of the joints under consideration, with the greatest changes in contact pattern and area being associated with the most mobile joints. CONCLUSION: The small change in absolute contact area with increased loading reflects the role of the foot, in providing a stable support for the body during gait. PMID- 11415761 TI - Impaired viscoelastic behaviour of spastic plantarflexors during passive stretch at different velocities. AB - OBJECTIVE: The aim of this study was to evaluate the effect of velocity on non reflexly induced resistive torque (RT) responses of the spastic plantarflexors (PFs) of subjects with spinal cord injuries. DESIGN: Descriptive study, transversal. BACKGROUND: In spastic muscles, non-reflex changes such as increased muscle stiffness, contractures and atrophy have been reported. These changes probably alter muscle tensile properties and viscoelastic behaviour. METHODS: Six subjects with chronic (1-3 yr) spinal cord injuries (SCI) and 12 normal controls (CTLs) participated in this study. Passive ankle dorsiflexions (DFs), ranging from -35 degrees to 5 degrees of DF, were randomly imposed at 5 degrees /s, 10 degrees /s, 20 degrees /s, 40 degrees /s, 60 degrees /s, 120 degrees /s and 180 degrees /s using an isokinetic dynamometer (Kin-Com(TM)). Unwanted muscle activity was detected using surface electrodes on the soleus and tibialis anterior muscles. RESULTS: The results show first that RT rose with increasing velocity; the RT increment reached statistical (analysis of variance (ANOVA); Scheffe post-hoc procedure) significance (P < 0.01) at a lower velocity for the CTLs (40 degrees /s) group than for the SCI group (60 degrees /s). Second, significantly (t-test; P < 0.001) larger net increments of RT (RT at each velocity minus RT at 5 degrees /s) were found for the CTLs at 180 degrees /s than for the SCI group whose RT plateaued at 60 degrees /s. Finally, whereas the RT velocity relationship was linear (r = 0.94) in the CTLs, that of the SCI group followed a power regression model (r = 0.85). CONCLUSIONS: These results show that the spastic PFs of the subjects with SCIs have an impaired velocity sensitive behaviour, especially at high velocities of stretch where greater resistance is expected. PMID- 11415762 TI - Alterations in gait resulting from deliberate changes of arm-swing amplitude and phase. AB - OBJECTIVE: The purpose of the study was to reach a better understanding of the influence of arm movement in human gait. DESIGN: The experiment involved the adoption of different arm-swing patterns by normal subjects who walked in a gait laboratory. BACKGROUND: The reciprocal swinging of the arms in walking plays an important role in gait, but the details require further research. More insight into the role of the arm-swing in gait could be obtained by adopting different arm-swing patterns while walking at different speeds. METHOD: Thirteen normal subjects were therefore requested to walk adopting different arm-swing patterns which included, one or both arms strapped to the body, full reciprocal excursion of both arms, pace walking and swinging the arms in parallel. From measurements made at different speeds of walking, regressions were calculated which made it possible to compare basal gait characteristics of the different arm patterns. RESULTS: Maximum velocity decreased in all cases. In the test conditions where the arms were strapped, the subjects increased velocity by increasing stride frequency more than stride length. In the other conditions the reverse was the case. CONCLUSION: The experiments show that the gait pattern is very much influenced by changes in arm-swing but the effects are related to the pattern of arm-swing and the speed of walking. PMID- 11415763 TI - Biomechanical characteristics of human trabecular bone. AB - OBJECTIVE: To evaluate the effect of strain rate (varepsilon), apparent density (rho(a)) and tissue density (rho(t)) on Young's modulus (E), strength (sigma(u)) and ultimate strain (varepsilon(u)) on intervertebral bone from a Chinese population. METHODS: Testing was performed by uniaxial compression at five strain rates on 36 human trabecular bone specimens from three male T(12) thick similar L(4) vertebrae. RESULTS: Apparent density ranged between 0.46 and 0.71 g/cm(3). Tissue density ranged between 1.02 and 1.54 g/cm(3). Non-linear regression analyses using strength, Young's modulus or ultimate strain as dependent variables (Y) and strain rate and apparent density or tissue density as independent variables were performed using the following equation: Y = arho(b)varepsilon(c). The exponent of apparent density and strain rate to Young's modulus were 1.88 and 0.07, separately (P = 0.0007). The variation of strength was explained only by apparent density with an exponent of 1.29 (P = 0.0107). The variation of Young's modulus was explained equally by the quadratic and cubic relationship to apparent density or tissue density (P < 0.01). Ultimate strain varied independently of apparent density or tissue density and strain rate. PMID- 11415764 TI - Biomechanical changes in the low back following reduction mammaplasty surgery. AB - OBJECTIVE: To give evidence of the mechanical consequences of reduction mammaplasty (RM) on the low back. DESIGN: A repeated-measures analysis was implemented to test the effect of RM on the external loads and angular velocity of the back during both static and dynamic lifting tasks. BACKGROUND: Patient follow-up surveys have documented a decrease in the frequency of low back pain following RM, but there is no quantitative data regarding biomechanical changes following surgery. METHODS: Patients were evaluated before and 4-8 weeks following RM. Flexion moment, compression and shear forces at L3-L4 were quantified for isometric flexion angles between 0 and 40 degrees. External loads and angular velocities of the back were studied during rapid dynamic lifting tasks. RESULTS: Isometric external flexion moments at L3-L4 decreased following RM. RM did not effect the applied flexion moment in the lumbar spine, but a trend suggested that RM resulted in increased lifting velocity. CONCLUSIONS: RM does act to reduce the loads on the lumbar spine during simple isometric tasks. During dynamic tasks, subjects may be able to lift faster without generating larger loads. PMID- 11415765 TI - The distribution of cartilage thickness in the knee-joints of old-aged individuals -- measurement by A-mode ultrasound. AB - OBJECTIVE: To provide data on the normal distribution of cartilage thickness in the knee joints of old-aged individuals. DESIGN: The accuracy and reproducibility of cartilage thickness measurements were evaluated with A-mode ultrasound, and the cartilage thickness distribution was examined throughout both knee joints of nine individuals aged between 62 and 94 yr. BACKGROUND: Data on the variation of cartilage thickness in the joint surfaces are relevant for the design of computer models of diathrodial joints and for surgical and arthroscopic procedures, particularly the calculation of cartilage material properties from indentation tests. METHODS: A 12.5 MHz A-mode ultrasound transducer was used, and after determining the accuracy and reproducibility of the system in comparison with CT arthrography and anatomical sections, the cartilage thickness was measured throughout the knee at 256 coordinate points. RESULTS: A-mode ultrasound yields results consistent with established methods. The cartilage thickness is distributed regularly in the knee-joint of older individuals. However, the absolute values are considerably lower than those previously reported for younger people, the mean (and maximal) thickness being 2.0 mm (3.3 mm) in the patella, 1.9 mm (3.3 mm) in the femur, and 1.9 mm (3.5 mm) in the tibia. CONCLUSION: If the results of computer models or surgical strategies are to be applied to the demographically important group of older individuals, the typical distribution of articular cartilage thickness in this group should be taken into consideration. PMID- 11415766 TI - Critical load of the human cervical spine: an in vitro experimental study. AB - OBJECTIVE: To determine the critical load of the osteoligamentous cervical spine in frontal plane. DESIGN: Whole human cervical spine specimens were loaded in axial compression with increasing force until the point of buckling. BACKGROUND: The osteoligamentous cervical spine and the surrounding muscles support the weight of the head and the external loads applied to it. Critical load is the maximum compressive force that the spinal column can sustain before buckling. Critical loads have been obtained for the osteoligamentous thoracolumbar spine (without the rib cage) and the lumbar spine. Critical load of the cervical spine has not yet been determined. METHODS: When a compressive force is applied to the cervical spine, it bends in the sagittal plane producing greater lordosis. The determination of critical load in Euler's sense requires blocking of this sagittal plane bending. A special apparatus was developed that constrained such bending in the sagittal plane, but allowed complete freedom of the spine motion in the frontal plane. Experiments were conducted to determine the axial force lateral bending curves of whole cervical spine specimens. Critical load values were obtained from these curves. As an alternative to this method, bending stiffness in the frontal plane was experimentally determined and the critical load was computed using Euler's theory of columns. RESULTS: Based upon the study of seven spine specimens (CO-T1), the critical load for the human cervical spine was found to be 10.5 (3.8) N obtained by direct experimentation. The average critical load calculated with the Euler theory using bending stiffness data, was 11.9 (2.0), but there were large individual differences when compared with the experimental results. CONCLUSIONS: The critical load of the osteoligamentous human cervical spine is about one-fifth to one-quarter the weight of the average head. PMID- 11415767 TI - Effects of a lumbar support on spine posture and motion assessed by electrogoniometer and continuous recording. AB - OBJECTIVE: To determine whether belt wearing causes changes in lumbar posture and motion during standing and work-related activities. DESIGN: The lumbar spine sagittal kinematics of healthy subjects were assessed with an electrogoniometer during a dynamic test and continuous recording with and without a lumbar support. Correlation between data from electrogoniometer and X-ray was established in preliminary experiments. BACKGROUND: The effects of a lumbar support were previously investigated in few patients using radiological or invasive techniques under laboratory conditions, with special regard to the restriction of global motion in flexion and extension. Whether lumbar posture may also be involved during belt wearing remains unclear. There is also no evidence that these changes affecting lumbar motion and posture persist during prolonged activities. METHODS: The correlation between electrogoniometer and radiographic data was assessed in 12 subjects. Lumbar curve angles were evaluated in 15 healthy subjects, with and without one type of lumbar support, during standing in orthostatic, fully-flexed and fully-extended positions and during work-related activities with a portable computer. Lumbar motion parameters were flexion, extension and total range of motion. Lumbar posture parameters were lumbar curve angle in orthostatic position and mean lumbar curve angle during continuous recording. RESULTS: Electrogoniometric and radiographic data from lumbar curve angles and motion changes during flexion/extension of the spine were slightly different but reasonably well correlated (r = 0.58-0.77). The lumbar support decreased the mean total range of motion of the lumbar spine during a single flexion/extension movement by 17% and during continuous recording by 22%. The lumbar curve angle in the orthostatic position was reduced by 3 degrees and the mean lumbar curve (assessed by continuous recording) was reduced by 4 degrees (mean for 15 subjects). There were major individual changes in these two parameters and the reduction in lumbar curvature was correlated with the initial (r = 0.66-0.72). CONCLUSION: Electrogoniometer data on lumbar motion are reasonably well correlated with X-ray data. The comparative values registered during flexion/extension tests and continuous recording confirm that a support belt limits slightly global lumbar motion. As changes affecting postural parameters (orthostatic or mean values during continuous recording) depend on subject morphometry, individual parameters should be taken in account when evaluating and using a lumbar support. PMID- 11415768 TI - Discrimination between maximal and submaximal effort in lumbar dynamometry. AB - OBJECTIVE: The purpose of this study was to investigate differences in consistency between maximum, preferred and a low level trunk performance. It was hypothesized that if maximum effort yielded consistent performances while submaximal efforts were less consistent, a consistency parameter could be used to detect lack of effort. DESIGN: Survey study. METHODS: Eleven healthy subjects performed lumbar dynamometry measurements (Isostation-B200) with maximum, preferred and low effort. A standard protocol was used and velocity and static torque were measured. Five subjects performed three measurements with low effort and trained the low effort level in between to determine the impact of training on consistency of submaximal performances. Coefficient of variations (CV), percentage differences and onset slopes were used to determine consistency. RESULTS: Results show that maximum and preferred effort are equally consistent. Maximum and low effort and preferred and low effort show small significant differences in CV for some velocity and isometric torque parameters. No significant differences between the three performance levels were indicated for percentage differences and onset slopes. Training did not improve the consistency of the measurements. CONCLUSIONS: Small significant differences in CV were found between maximal and submaximal effort. However the combination with the relatively large standard deviations make it unlikely that this parameter can be used for a reliable discrimination. PMID- 11415769 TI - Comparison between two dynamic methods to estimate triaxial net reaction moments at the L5/S1 joint during lifting. AB - OBJECTIVE: The purpose of the study was to evaluate the validity of two biomechanical dynamic tridimensional (3D) multisegment models. DESIGN: An experimental study was conducted for this purpose. BACKGROUND: The models that were compared are commonly used to analyse lifting tasks. The upward model used the measured external forces at the feet, whereas the downward model used an estimation of the forces at the hands. The downward model approach is more practical to study manual materials handling (MMH) tasks but needs improvement to produce valid estimates. METHODS: One male subject participated in the study. Four cameras and two force platforms were used to get the data from 66 static and 108 dynamic trials. The triaxial net moments at L5/S1 of both models were compared. RESULTS: Good agreement was observed between the two models for the moments of the static tasks but great disparity was observed for the moments of the dynamic tasks (up to 78 Nm in extension). Both models were sensitive to different task parameters (movement or load asymmetry, movement rate, load magnitude). CONCLUSIONS: The direct measurement of the external forces at each hand is recommended. The validation should be performed with more subjects for generalization. PMID- 11415770 TI - The effects of cyclic stretching on tensile properties of the rabbit's skeletal muscle. AB - OBJECTIVE: To define the threshold of muscle injury with cyclic passive stretch. DESIGN: The changes in the load-deformation curve of muscle-tendon unit were monitored until the failure point by an in vivo rabbit model. BACKGROUND: Muscle injuries range in severity from a simple strain to complete rupture. Although strains occur more frequently than complete failures, only a few studies have investigated the phenomena of these sub-failure injuries. Monitoring of the continuum for stretch-induced injury allows us to define the threshold of stretch injury. METHODS: Thirty rabbits' triceps surae muscle-tendon unit preparations were used. One of the pairs (control) was stretched until failure; the other (experimental) was first cyclic stretched to either 12, 20 or 25% of the initial length of the muscle-tendon unit and then stretched to failure. Comparisons were made between the load-deformation curves of the experimental and control specimens. RESULTS: When cyclic stretched to 12 or 20%, there were no significant changes existed in the biomechanical parameters except the deformation at the peak load. In contrast, all the biomechanical parameters except the ration of the energy absorption changed significantly after 25% strain cyclic stretch. CONCLUSIONS: A threshold for stretch-induced injury does exist. This can be reproduced at the 25% strain of the triceps surae muscle-tendon unit. PMID- 11415771 TI - Contribution of capsuloligamentous structures to passive static inferior glenohumeral stability. AB - OBJECTIVE: This study attempted to determine the relative stabilizing effect of the capsuloligamentous structures to inferior humeral displacement. DESIGN: This was an in vitro study, utilizing cadaveric shoulder specimens. BACKGROUND: Inferior glenohumeral instability has come under increasingly close scrutiny in the literature in recent years, yet a description of the precise pathoanatomy is still lacking. METHODS: Thirteen shoulder specimens were cleaned of soft tissue superficial to the vented joint capsule. The position of the humeral head relative to the glenoid was determined. The humeri were subjected to an inferior load. Static position recordings were obtained for each specimen: (1) with the humerus adducted and abducted, (2) before and after sectioning the superior and inferior capsuloligamentous structures. These recordings were analyzed. RESULTS: In adduction, the humeral head migrated inferiorly as the capsuloligamentous structures were sectioned, but no capsuloligamentous structure was seen to be most important. In abduction, when the inferior capsuloligamentous structures were sectioned first, the humeral head migrated inferiorly (P<0.001). When the superior structures were sectioned first, the humeral head position did not significantly change. CONCLUSIONS: In this experimental model, the inferior capsuloligamentous structures are the primary inferior stabilizers of the abducted shoulder. The primary stabilizers of the adducted shoulder remain unclear. PMID- 11415772 TI - Biomechanics of the hindfoot joints in response to degenerative hindfoot arthrodeses. AB - OBJECTIVE: The aim of this study is to understand how the different single and combined hindfoot arthrodeses affect the biomechanics of the hindfoot. DESIGN: The range of rotation of each hindfoot bone in in vitro specimens were measured and compared before and after prescribed arthrodeses. BACKGROUND: Single and combinations of single arthrodeses in the hindfoot are used by surgeons to correct hindfoot deformities. Clinical reports of both successful and less successful procedures are reported in the clinical literature, backed by unfounded statements as to the biomechanics of the hindfoot. METHODS: Eight fresh frozen specimens were mounted in a frame and loaded to induce maximal dorsiflexion, plantarflexion, and rotation about the long axis of the foot, defined as supination and pronation. The three dimensional motion of the talus, calcaneus, navicular, and cuboid was tracked using an ultrasonic positioning system. The resulting range of rotation (RoR) of each bone in dorsiflexion to plantarflexion and supination to pronation about the calculated helical axis was then compared. Arthrodeses were then systematically introduced and the measurements repeated. RESULTS: Significant differences in RoR were found at each joint corresponding to specific combinations of arthrodeses, P < 0.05. At the calcaneocuboid joint, statistically normal rotations were calculated when the subtalar joint was arthrodized. At both the talonavicular and subtalar joints, statistically normal rotations were found when the calcaneocuboid joint was fused. In all of the other arthrodesis conditions, statistically stable joints were found. CONCLUSIONS: The results show that the mobility of each hindfoot joint is dependent on the mobility of the other hindfoot joints, and that the function of the talonavicular joint has the greatest influence on overall hindfoot function. PMID- 11415773 TI - The relative skin movement of the foot: a 2-D roentgen photogrammetry study. AB - OBJECTIVE: This paper describes a validation of the relative motions of skin mounted markers on the foot. DESIGN: The movement of a skin-mounted marker on the foot was evaluated by the use of 2-D roentgen photogrammetry. BACKGROUND: Validations of skin-mounted markers are needed in order to assess whether clinical use of motion analysis systems is accurate. METHODS: Six healthy volunteers participated in the study. Measurements were made with 2-D roentgen photogrammetry from the medial aspect of the foot. Spherical lead markers, with a diameter of 2 mm, were glued on the skin over the following landmarks: the medial malleolus, the navicular bone, the medial part of calcaneus, the base and the head of the first metatarsal bone, and the base of the fifth metatarsal bone. The foot was placed on a platform that was adjustable in three directions: horizontal, 20 degrees upward (towards dorsal flexion), and 30 degrees downward (towards plantar flexion). RESULTS: In relation to the underlying bones, the markers were found to move up to 4.3 mm. The markers with the largest movements were the ones over the medial malleolus, the navicular bone and the calcaneus. For the distally placed markers i.e. at the base and the head of the first metatarsal bone, and the base of the fifth metatarsal bone, the displacements were a maximum 1.8 mm. CONCLUSIONS: Markers mounted on the foot moved between 1.8 and 4.3 mm corresponding to the underlying bones. The most proximal attached markers demonstrated the largest movements. PMID- 11415774 TI - The 3-D motion of the centre of gravity of the human body during level walking. I. Normal subjects at low and intermediate walking speeds. AB - OBJECTIVE: To measure the mechanical energy changes of the centre of gravity (CG) of the body in the forward, lateral and vertical direction during normal level walking at intermediate and low speeds. DESIGN: Eight healthy adults performed successive walks at speeds ranging from 0.25 to 1.75 m s(-1) over a dedicated force platform system. BACKGROUND: In previous studies, it was shown that the motion of the CG during gait can be altered more than the motion of individual segments. However, more detailed normative data are needed for clinical analysis. METHODS: The positive work done during the step to accelerate the body CG in the forward direction, W(f), to lift it, W(v), to accelerate it in the lateral direction, W(I), and the actual work done by the muscles to maintain its motion with respect to the ground ('external' work), W(ext), were measured. This allowed the calculation of the pendulum-like transfer between gravitational potential energy and kinetic energy of the CG, (percentage recovery, R). At the optimal speed of about 1.3 m s(-1), this transfer allows saving of as much as 65% of the muscular work which would have been otherwise needed to keep the body in motion with respect to the ground. The distance covered by the CG at each step either forward (step length, S(I)), or vertically (vertical displacement, S(v)) was also recorded. RESULTS: W(I) was, as a median, only 1.6-5.9% of W(ext). This ratio was higher, the lower the speed. At each step, W(ext) is needed to sustain two distinct increments of the total mechanical energy of the CG, E(tot). The increment a takes place during the double stance phase; the increment b takes place during the single stance phase. Both of these increments increased with speed. Over the speed range analyzed, the power spent to to sustain the a increment was 2.8-3.9 times higher than the power spent to sustain the b increment. PMID- 11415775 TI - The 3-D motion of the centre of gravity of the human body during level walking. II. Lower limb amputees. AB - OBJECTIVE: To analyse the motion of the centre of gravity (CG) of the body during gait in unilateral lower limb amputees with good kinematic patterns. DESIGN: Three transtibial (below-knee, BK) and four transfemoral (above-knee, AK) amputees were required to perform successive walks over a 2.4 m long force plate, at freely chosen cadence and speed. BACKGROUND: In previous studies it has been shown that in unilateral lower limb amputee gait, the motion of the CG can be more asymmetric than might be suspected from kinematic analysis. METHODS: The mechanical energy changes of the CG due to its motion in the vertical, forward and lateral direction were measured. Gait speed ranged 0.75-1.32 m s(-1) in the different subjects. This allowed calculation of (a) the positive work done by muscles to maintain the motion of the CG with respect to the ground ('external' work, W(ext)) and (b) the amount of the pendulum-like, energy-saving transfer between gravitational potential energy and kinetic energy of the CG during each step (percent recovery, R). Step length and vertical displacement of the CG were also measured. RESULTS: The recorded variables were kept within the normal limits, calculated in a previous work, when an average was made of the steps performed on the prosthetic (P) and on the normal (N) limb. Asymmetries were found, however, between the P and the N step. In BK amputees, the P step R was 5% greater and W(ext) was 21% lower than in the N step; in AK amputees, in the P step R was 54% greater and W(ext) was 66% lower than in the N step. Asymmetries were also found in the relative magnitude of the external work provided by each lower limb during the single stance as compared with the double stance: a marked deficit of work occurred at the P to N transition. PMID- 11415776 TI - Age-related changes in the ability to side-step during gait. AB - OBJECTIVE: The purpose was to identify age-related changes in the ability to step sideways during gait. DESIGN: Sixteen young women (mean age 27) and 16 older women (mean age 70) were tested. BACKGROUND: The ability to safely incorporate changes of direction into our walking pattern at short notice is a requirement for full mobility. Side-stepping in particular is frequently used to avoid obstacles. METHODS: The task was to walk repeatedly down the center of a walkway divided into three parallel lanes. A suddenly-appearing visual cue instructed the subject to move to either the right or left lane or to remain in the center lane. The aim was to make the shift accurately but as quickly as possible, without halting forward progression. RESULTS: In only 26% of the trials by the older women was the shift accomplished by taking just one extra step, compared with 58% of the trials by the young women. For one of the two shift directions there was no significant difference between the groups in either speed changes or foot placement accuracy. For the other direction, however, the young women incorporated the shift with no significant change in average speed; the older women's speed decreased by 3.3%. The older women also made more errors in foot placement in the steps following the side-step (a 22% error rate compared to a 3% error rate). CONCLUSIONS: Older women were less able to quickly and accurately incorporate a sideways shift in the plane of progression into their comfortable walking pattern. PMID- 11415777 TI - Theoretical analysis of ligament and extensor-mechanism function in the ACL deficient knee. AB - OBJECTIVE: To study ligament and extensor-mechanism function in the ACL-deficient knee. DESIGN: Mathematical modeling of the muscles, ligaments, and bones at the knee. BACKGROUND: Numerous experiments have documented an increase in anterior tibial translation (ATT) in the ACL-deficient knee, but its effect on the function of the knee-extensor mechanism is not fully understood. The load sharing between the knee ligaments is also unknown since ligament forces are difficult to measure in vivo. METHODS: The geometry of the model bones is adapted from cadaver data. Eleven elastic elements describe the geometric and mechanical properties of the ligaments and joint capsule. The model is actuated by eleven musculotendinous units. Straight, anterior drawer and maximum, isometric extension are simulated by solving the equations for static equilibrium of the model. RESULTS: The moment arm of the extensor mechanism and the torque at the knee are nearly equal in the intact and ACL-deficient model. Knee-ligament forces are lower in the ACL deficient model than in the intact model. Ligament forces are lower because the shear force applied to the tibia decreases when the model ACL is removed. CONCLUSIONS: Function of the knee-extensor mechanism is not altered by loss of the ACL. The MCL is the primary restraint to anterior drawer in the ACL-deficient knee. The deep fibers of the MCL dominate the load sharing between the ligaments when the ACL is absent. PMID- 11415778 TI - The mechanical properties of human alar and transverse ligaments at slow and fast extension rates. AB - OBJECTIVE: To study biomechanics and failure characteristics of the alar and transverse ligaments of the upper cervical spine at two different extension rates. DESIGN. IN VITRO: biomechanical study using human and alar and transverse ligament specimens. BACKGROUND: Previous studies of these ligaments have been at relatively slow extension rates, even though the real life trauma occurs at significantly higher extension rates. METHODS: Fresh human cadaveric alar and transverse ligaments were subjected to slow (0.1 mm/s) and then fast (920 mm/s) extension rates. Force and elongation curves were recorded during the testing, and several biomechanical parameters were computed and compared. First the specimen was stretched at the slow extension rate up to 70 N for the alar ligament and 140 N for the transverse ligament. Then the specimens were stretched until failure at the fast extension rate. RESULTS: Average initial lengths of the alar and transverse ligaments were 11.2 and 18.0 mm, respectively. At 70 N, the average alar ligament strain decreased from 16.4 to 0.3%, the stiffness increased from 80 to 2316 N/mm and the energy absorbed decreased from 47.4 to 1.3 Nmm, as the extension rate increased from the slow to the fast. At failure, the average strain, force and energy absorbed at the fast extension rate were respectively 3.1%, 367 N and 123 Nmm. For the transverse ligament at 140 N, the strain decreased from 12.5 to 0.6%, the stiffness increased from 141 to 1472 N/mm, and the energy absorbed decreased from 96.1 to 7.6 Nmm as the extension rate went from slow to the fast. At failure, strain, force and energy absorbed for the transverse ligament at the fast extension rate were, respectively, 2.3% and 436 N and 101 Nmm respectively. CONCLUSION: Within the physiological limits, the strain and energy absorbed decreased to less than one tenth, while the stiffness increased to greater than ten times as the extension rate increased, for both the alar and transverse ligaments. When failed at the faster rate, the alar ligament, although weaker of the two, absorbed greater energy to failure because of its higher failure strain. PMID- 11415779 TI - Coordination between the lumbar spine lordosis and trunk angle during weight lifting. AB - OBJECTIVE: To analyze the coordination of the lumbo-sacral angle (lumbar spine lordosis) and the trunk inclination during lifting of different loads. STUDY DESIGN: Kinematic data of spine motion were analyzed. The parameters characterizing the relationships between the lordosis and the trunk inclination angle were estimated. BACKGROUND: The shape of the spine has been analyzed mostly for static or quasi-static conditions. The parameters relating the lumbar spine lordosis and trunk inclination in dynamics have not been analyzed. METHODS: Healthy subjects performed unconstrained weight lifts from ground to mid-thigh level. Kinematic data were derived from the tracking of markers (light-emitted diodes) placed over the spine and pelvis using an OPTPTRAK system. The relationship between lordosis and trunk inclination was analyzed. RESULTS: The relationship between lumbar spine curvature (lumbo-sacral angle or lordosis) and trunk inclination during weight lifting was described by an exponential function with three parameters. These were the lordosis extremes associated with the horizontal and vertical positions of the trunk and the trunk inclination when lordosis equals zero. The absolute value of the lordosis angle decreases at the onset of the extension phase of lifting when the load increases, implying active reaction of musculosceletal system to increasing load. CONCLUSIONS: The changes in the lordosis and trunk inclination are strictly correlated implying that the nervous system actively coordinates the degrees of freedom of the spine, providing an inter-joint synergy. PMID- 11415780 TI - A new method for measuring wrist-joint ligament length changes during sagittal and frontal motion. AB - OBJECTIVE: The present study addresses elongation patterns of selected capsular wrist ligaments during two types of planar wrist motion. DESIGN. IN VITRO: continuous measurement of capsular ligament elongation using miniature displacement transducers. BACKGROUND: The function of the capsular wrist ligaments in carpal kinematics is still to be determined unambiguously. Most previous studies on carpal ligament behaviour were realised using X-ray stereophotogrammetric methods. METHODS: Elongation of the palmar radiolunate, radiocapitate and ulnocapitate ligaments, as well as of the dorsal radiotriquetral ligament during frontal and sagittal wrist motion were analysed in 10 anatomical specimens using miniature differential variable reluctance transducers. Wrist motion was tracked using a six-degree-of-freedom electrogoniometer. RESULTS AND CONCLUSIONS: In most instances, the elongation of these ligaments was similar for two motion modalities tested (guided by a rail or free) with, however, a trend to larger individual variations and smaller elongation during free motion for several ligaments. The capsular ligaments considered in this study displayed length changes inferior to 15% of the initial length. The individual elongation patterns and ranges were highly variable, contrasting with excellent intra-individual reproducibility. PMID- 11415781 TI - The measurement of muscle stiffness in anterior cruciate injuries -- an experiment revisited. AB - OBJECTIVE: To investigate the reported changes in angular stiffness of the knee due to the hamstrings muscles in the anterior cruciate ligament (ACL) deficient knee. DESIGN: The flexors of the knee were modelled as springs and the model tested on patients and controls. BACKGROUND: A previous study had used a similar model but showed no difference between an ACL deficient and an uninjured knee. METHODS: A modification of a recently described method for measuring the stiffness of the hamstrings was used. ACL deficient patients were investigated and also normal controls. RESULTS: We found that stiffness not only varied between patients, but also between legs, with the injured leg having significantly stiffer flexor muscles (P < 0.001). We also showed that the stiffness of the knee due to the flexors is the same in normal subjects as that in the uninjured contralateral knee of the ACL injured patient. CONCLUSIONS: The measurement of the stiffness of the hamstrings following ACL injury may be a valid method of assessing patients with such injuries. Further investigation is required. PMID- 11415782 TI - Evaluation of spinal loading during lowering and lifting. AB - OBJECTIVE: To estimate the three-dimensional spinal loads during various lifting and lowering tasks. DESIGN: The in vivo measurements of the trunk dynamics, moments, and myoelectric activity were used as inputs into an electromyographic assisted model used to predict the three-dimensional spinal loads. BACKGROUND: Previous studies of eccentric motions have investigated muscle activity, trunk strength, and trunk moments. A void in the body of knowledge exists in that none of these studies investigated spinal loading. METHODS: Ten subjects lifted (40 degrees of flexion to 0 degrees ) and lowered (0 degrees of flexion to 40 degrees ) boxes while positioned in a structure that restrained the pelvis and hips. The tasks were performed under isokinetic trunk velocities of 5, 10, 20, 40, and 80 deg s(-1) while holding a box with weights of 9.1, 18.2, and 27.3 kg. RESULTS: Lowering strength was found to be 56% greater than lifting strength. The lowering tasks produced significantly higher compression forces but lower anterior posterior shear forces than the lifting tasks. The differences in the spinal loads produced by the two lifting tasks were attributed to differences in coactivity and unequal lifting moments (i.e. holding the box farther away from the body). CONCLUSIONS: The nature of the spinal loads that occur during lowering and lifting were significantly different. The difference in spinal loads may be explained by different lifting styles. PMID- 11415783 TI - Control of knee stability after ACL injury or repair: interaction between hamstrings contraction and tibial translation. AB - OBJECTIVE: The aim of this study was to investigate the interaction between hamstrings contraction and tibial translation during isometric quadriceps contraction (IQC) and their effects on cruciate ligament forces. METHODS: A mathematical model of the knee in the sagittal plane was used with simple geometric representation for the model muscles, ligaments and bones. A mechanical analysis was used to calculate muscle, ligament and contact forces. RESULTS: The results show that isolated IQC requires a posterior force applied to the tibia at the knee over the range 0-85 degrees flexion, usually applied by the anterior cruciate ligament (ACL). The model hamstrings can provide this posterior force, except near extension, resulting in reduced anterior tibial translation (ATT) and, therefore, reduced ACL force. ATT varied inversely with the hamstrings force. The interaction between the two was highly dependent on flexion angle, the hamstrings force being least effective in reducing ATT near extension. CONCLUSIONS: Flexing loads placed distally on the tibia can stretch an injured or repaired ACL. The analysis suggests that modest hamstrings forces during IQC can help protect the ACL. In the complete absence of an ACL, hamstrings contraction as well as ATT may be inevitable while using quadriceps near extension. PMID- 11415784 TI - Elongation patterns of the collateral ligaments of the human knee. AB - OBJECTIVE: To determine the responses of the medial and lateral collateral ligaments (MCL, LCL) of the human knee to externally applied stresses. DESIGN: Differential variable reluctance transducers were used to measure length changes along the long posterior parallel fibers of the MCL and the middle third of the LCL through a flexion range of 15-120 degrees and a variety of external stresses. BACKGROUND: There is a lack of consensus regarding the collateral ligament response to internal and external tibial rotation. In addition, there are very few studies that have investigated the effects of quadriceps and hamstrings muscle group loads on the strain in the collateral ligaments. METHODS: Three series of tests were performed. First, the passive behavior of the ligaments was obtained as well as the ligament response to 3 degrees of varus and valgus rotation. The next series tested the ligaments' response to 0-10 degrees of internal and external tibial axial rotation. Finally, isolated and co-contracted quadriceps and hamstrings muscle group loads were applied. RESULTS: The instrumented portions of both ligaments were more strained in extension than flexion. Varus rotations stretched the LCL, whereas valgus rotations elongated the MCL. The strain in the MCL was shown to increase during external rotation and decrease during internal rotation. The LCL did not exhibit a uniform response across specimens to internal or external tibial axial rotation, but was consistent between left and right knees from a single cadaver. Highly consistent trends of interactions between muscle loads and the strain in both the MCL and LCL were noted. CONCLUSIONS: The responses of the MCL and LCL to applied stresses are dependent upon the flexion angle of the knee, the influence of muscle loading and, to a lesser extent, anatomic variation in the ligaments themselves. Guidelines for rehabilitation of the collateral ligaments following injury are suggested. PMID- 11415785 TI - Rising from chair after total knee arthroplasty. AB - OBJECTIVE: This study was performed to determine the biomechanics of chair rising by patients after successful total knee arthroplasty (TKA). DESIGN: Relative full body joint positions and ground reaction forces were measured by a motion analysis system and two force plates. BACKGROUND: Chair-rise produces increased joint forces and moments compared with level walking, and it is difficult to rise from a chair for most patients with neurological or musculoskeletal abnormalities. Previous motion studies of patients after TKA had focused on gait analysis and stair climbing. METHODS: Twelve patients after successful TKA were studied while performing sit-to-stand transfer from a chair at four chair heights without the use of arm rests. The results of this group were compared with those of 12 healthy elderly subjects and 14 osteoarthritic patients before TKA. RESULTS: Compared with the healthy elderly group during chair rising, the patients after TKA had increased horizontal mass center velocity, increased vertical hip joint forces, the maximum sound-side hip extension moment, but decreased vertical mass center velocity. CONCLUSIONS: Compensatory mechanisms of chair rising were adopted by the arthritic patients before and after TKA. The mechanisms include increased forward body bending and more weight shift on the sound side. PMID- 11415786 TI - Dynamic joint analysis as a method to document coordination disabilities associated with Parkinson's disease. AB - OBJECTIVE: The purpose of the present study was to introduce dynamic joint analysis and subsequent phase partitioning of the movement pattern as a method for investigating the force control deficits associated with Parkinson's disease. DESIGN: Pilot data were collected from four non-impaired individuals and four patients afflicted with Parkinson's disease while performing arm movements of different spatiotemporal features. BACKGROUND: Investigation of motor performance in Parkinson's patients has related the clinically observed symptoms to the ability to control muscular force. METHODS: Experimental movements were filmed using a high speed camera operating at 200 Hz. The mechanical power characteristics of the elbow and shoulder were determined by applying inverse dynamic solutions to the kinematic data. Movement was reflected in a series of goal-directed phases describing the functional role of muscle activity across the joint. RESULTS: The analysis revealed that Parkinson's disease impairs the ability of the muscles to produce the energy required for performing ballistic, segmental movements. Moreover, patients demonstrated greater difficulty in controlling a two-joint task as opposed to a single-joint one; this was reflected by additional phases of activity at the elbow. CONCLUSIONS: Slowness of movement is associated with the inappropriate scaling of the muscle force, as well as with the limited use of motion-dependent forces in accelerating distal segment movement. PMID- 11415787 TI - The windlass mechanism during normal walking and passive first metatarsalphalangeal joint extension. AB - OBJECTIVE: To assess the relationship between passive first metatarsophalangeal extension and rearfoot motion during walking on the windlass mechanism. DESIGN: An in vivo study of the windlass mechanism during passive extension of the first metatarsophalangeal joint and walking. BACKGROUND: Despite theoretical work regarding the windlass mechanism in cadavers, there is little research concerning its in vivo role. METHODS: The amount of first metatarsophalangeal extension and medial longitudinal arch movement of the right foot in 20 subjects between the age of 22 and 55 years was recorded as they sat with their right hip, knee and ankle in 90 degrees and their great toe passively extended. In addition, movement of the rearfoot was recorded while they walked. RESULTS: The results of this study showed that the windlass mechanism is active during passive extension of the great toe. In addition, subjects could be divided into two groups (IMMEDIATE and DELAYED) based upon the timing of when the windlass mechanism began relative to passive extension of their first metatarsophalangeal joint. During walking, the DELAYED group was more inverted at heel strike and had a greater magnitude of rearfoot eversion. CONCLUSIONS: Two distinct subpopulations of individuals can be identified by when the windlass mechanism is initiated relative to passive first metatarsalphalangeal extension. PMID- 11415788 TI - A comparison of three-dimensional lower extremity kinematics during running between excessive pronators and normals. AB - OBJECTIVE: The purpose of this research was to compare the three-dimensional kinematics of runners exhibiting excessive rearfoot pronation with those having normal rearfoot pronation. DESIGN: The study design was a comparative investigation of two types of running patterns. BACKGROUND: Excessive rearfoot pronation is often linked with overuse injuries of the lower extremity. However, the literature is void of papers describing the rearfoot motion of runners presenting with excessive rearfoot pronation. Many knee-related injuries in runners are associated with increased rearfoot pronation; however, knee mechanics in this population of runners have yet to be studied. Finally, three-dimensional studies are needed to describe joint motion fully during running and these are also lacking. METHODS: Eighteen subjects (nine excessive pronators -- PRs; nine normals -- NLs) were studied during treadmill running at 3.35 m/s. Retroreflective markers were placed on the foot, shank and thigh segments and recorded with four 200 Hz video cameras. Three-dimensional kinematics were computed. RESULTS: A downward shift of the eversion curve was seen in the PR group resulting in an everted position of the rearfoot at both footstrike and toe off compared with an inverted posture seen in the NL group. The amount of toe-out was not significantly different between the two groups. At the knee, the PR group demonstrated significantly less adduction and significantly greater flexion than the NL. Mean peak velocities of the PR group were greater in all angular measures except knee adduction. However, only foot dorsiflexion and eversion and knee flexion velocities were significantly different. CONCLUSIONS: Kinematic differences were noted at both the rearfoot and the knee of the runners who exhibit excessive rearfoot pronation. PMID- 11415789 TI - Data management in gait analysis for clinical applications. AB - OBJECTIVE: To study the reliability of gait analysis data obtained using the Calibrated Anatomical System Technique (CAST) protocol and to verify the suitability and repeatability of the extraction of a number of parameters from the waveforms obtained. DESIGN: The experimental protocol and the parametric analysis technique were applied on a population of able-bodied subjects. BACKGROUND: The clinical interpretation process of gait data still needs a more accurate analysis of the reliability and repeatability of the measurements and a suitable procedure for data reduction useful for data comparison. METHODS: Gait analysis was performed in 20 able-bodied subjects using a stereophotogrammetric system and a forceplate. 124 parameters relative to time-distance, kinematic and kinetic variables were calculated by means of an automatic procedure and statistically analysed. RESULTS: Most of the parameters were found to be normally distributed with relatively small range of variation. Few of them showed poor repeatability, mostly due to the experimental inaccuracies introduced. Correlation of several gait parameters with age, sex, and speed of progression was also identified. PMID- 11415790 TI - Design and test of neural networks and statistical classifiers in computer-aided movement analysis: a case study on gait analysis. AB - OBJECTIVE: To describe the methods for designing and testing diagnostic systems in movement analysis and to verify the clinical usefulness of neural networks and statistical classifiers in a case study. DESIGN: Connectionist and statistical models trained and tested with measured data. BACKGROUND: A basic need in rehabilitation and related fields is to efficiently manage the vast information obtained from a movement analysis laboratory. Many studies have dealt with the interpretation of measured variables in order to correlate objective descriptors to the presence and/or severity of specific neuromusculoskeletal disorders or their consequences. This traditional analytical approach has been complemented in the last decade by new non-linear classification tools called neural networks. METHODS: A gait analysis study on 148 lower limb arthrosis patients and 88 age matched control subjects. Pathological and healthy gait patterns obtained from force plates wer discriminated by means of multilayer perceptrons and statistical classifiers. RESULTS: Ten input features were enough to train a multilayer perceptron with six hidden neurons. The discrimination rate of the neural net was 80% after cross-validation, significantly higher (P<0.05) than the performance of a Bayes quadratic classifier (about 75%). A great variance due to a small cross validation set could be demonstrated. CONCLUSIONS: Strict statistical requirements must be observed for designing a neural network. Although these models attain a better performance than conventional statistical approaches, the benefits they bring are sometimes not sufficient to justify their use. Furthermore, clinicians routinely involved in critical decisions may not consider such diagnostic systems reliable enough. PMID- 11415791 TI - Strength of internal fixation for calcaneal fractures. AB - OBJECTIVE: To compare the strength of two types of fixation method for calcaneal fractures. DESIGN: A biomechanical testing examined the stability of 12 fractured calcaneal specimens fixed with two different methods. BACKGROUND: Though anatomic reduction and internal fixation for the treatment of intra-articular fractures of the calcaneus has become popular, biomechanical data on the fixation strength is lacking. METHODS: Twenty fresh frozen specimens of amputated human legs were impacted by a 20 kg weight dropped from a 155 cm height to create calcaneal fractures. Twelve specimens which demonstrated a longitudinal and a transverse primary fracture lines were selected for open reduction and internal fixation. Group 1: a lateral buttress plate and parallel screws placed in the latero-medial direction were used. Group 2: a longitudinal screw was added in addition to the fixation used in group 1. Biomechanical testing was performed by applying a tibial shank load until the internal fixation failed. RESULTS: All mechanical failures of the reconstructed calcaneus occurred through the transverse primary fracture line. The average failure load was 805+/-356 N in group 1 and 2905+/-910 N in group 2 (Wilcoxon p<0.05). CONCLUSIONS: A longitudinal transfixing screw could significantly improve the stability of the transverse primary fracture line in calcaneal fractures. PMID- 11415792 TI - An assessment of wrist splint and glove use on wheeling kinematics. AB - OBJECTIVE: The purpose of this investigation was to assess whether glove and/or splint use could effectively reduce hyperextension at the wrist during wheelchair propulsion, thus potentially reducing the conditions predisposing to median nerve dysfunction, and to evaluate the overall effects of these interventions on wheeling mechanics. DESIGN: This investigation used a randomized experimental design. BACKGROUND: The upper extremities are used for weight bearing and propulsion by individuals who are wheelchair dependent. High intracarpal pressures created by hyper-extension of the wrist and repetitive high force stresses of the hands against the wheel are suggested causes of median nerve dysfunction. METHODS: The wheeling performances of 13 subjects were recorded using two SVHS video-cameras under four different glove/splint conditions. Each subject was analysed on two wheeling cycles under all four conditions. Wrist and elbow angles, joint range of motions and wheeling speed were determined. Data were analysed using a one-way analysis of variance (ANOVA), followed by Scheffe post-hoc comparisons at the 0.05 level of significance. RESULTS: The splint and the glove/splint combination significantly reduced wrist extension during wheeling, but did not alter elbow motion or maximal wheeling speed. CONCLUSIONS: Hand and wrist protection in the form of a splint or glove/splint combination can reduce hyperextension of the wrist, and therefore may be of functional value for wheelchair users. This form of protection may therefore be useful in various orthopedic conditions of the wrist or hand commonly seen in wheelchair-dependent individuals without seriously interfering with wheeling quality. PMID- 11415793 TI - Mechanism of whiplash injury. AB - OBJECTIVE: To propose a different hypothesis of whiplash injury mechanism based on a series of experimental studies summarized in this communication. DESIGN: A series of biomechanical studies simulating whiplash trauma using isolated human cadaveric spine specimens. BACKGROUND: Whiplash injuries are on the rise as reported in several recent studies, due primarily to the increased traffic density. Although the symptoms associated with whiplash have been described, our understanding of the injury mechanism remains poor. The prevailing view of neck hyper-extension causing the injury has not been supported by recent experimental studies. METHODS: Eight fresh human cadaveric cervical spine specimens were prepared and traumatized to varying degrees under controlled conditions using a bench-top model of whiplash trauma. Before and after each trauma, the specimen was studied by functional radiography and flexibility test to document changes in the anatomic alignment and biomechanical properties at each level indicating injuries sustained. At the end of all testing, CT-scans, MRI and cryomicrotome images were obtained. During each trauma, relative motions of all intervertebral joints were recorded with a high speed movie camera. Elongations of the vertebral artery and several capsular ligaments were also monitored during the trauma using specially designed transducers. RESULTS: The hyper-extension hypothesis of injury mechanism was not supported by these studies. We found a distinct bi-phasic kinematic response of the cervical spine to whiplash trauma. In the first phase, the spine formed an S-shaped curve with flexion at the upper levels and hyper extension at the lower levels. In the second phase, all levels of the cervical spine were extended, and the head reached its maximum extension. The occurrence of anterior injuries in the lower levels in the first phase was confirmed by functional radiography, flexibility tests and imaging modalities. The largest dynamic elongation of the capsular ligaments was observed at C6-C7 level during the initial S-shaped phase of whiplash. Similarly, the maximum elongation of the vertebral artery occurred during the S-shape phase of whiplash. CONCLUSION: We propose, based upon our experimental findings, that the lower cervical spine is injured in hyperextension when the spine forms an S-shaped curve. Further, this occurs in the first whiplash phase before the neck is fully extended. At higher trauma accelerations, there is a tendency for the injuries to occur at the upper levels of the cervical spine. Our findings provide truer understanding of whiplash trauma and may help in improving the diagnosis, treatment, and prevention of these injuries. PMID- 11415794 TI - Relationship between the conformity and the lubricating ability of synovial joints. AB - OBJECTIVE: To relate conformity of sliding surfaces with the lubricating ability of synovial joints. DESIGN: Measurement of start-up friction in the stifles of various animals. Assessment of conformity by Hertzian contact area. BACKGROUND: Past studies showed that the start-up friction in synovial joints sharply increased with the loading duration. The reasons why the friction increased and why the increasing rate is different in different joints were, however, not found. METHODS: Nine stifle joints from various animals were used. A robotic arm was used to give the compressive force and the sliding motion to the joint. Start up friction was measured by a universal force sensor. The principal curvatures of the sliding surfaces were directly measured by a radius-gauge. Hertzian contact area was calculated from the principal curvatures of the sliding surfaces. RESULTS: The duration until the frictional coefficient reached 0.1 was related to the Hertzian contact area. CONCLUSION: The conformity of sliding surfaces is related to the lubrication ability in synovial joints. The squeeze-film mechanism plays an important role in joint lubrication. PMID- 11415795 TI - Revised planimetric model of unipennate skeletal muscle: a mechanical approach. AB - OBJECTIVE: Planimetric models which are simple, in the sense that small numerical effort is needed, are used to study functional consequences of skeletal muscle architecture. This paper argues with the approach to derive force of a unipennate muscle based on only equilibrium of the aponeurosis (tendon-sheet). In such an approach intramuscular pressure gradients are neglected and no suitable aponeurosis force can be determined. METHOD: The approach presented in this paper is based on mechanical equilibrium of whole muscle. A volume-related force is introduced to keep muscle volume constant. Mechanical equilibrium of whole muscle yields a different relation between fiber and muscle force as well as length changes as a consequence of pennation, compared with relations derived when only equilibrium of aponeurosis is considered. RESULTS: The newly derived relation improved prediction of the rat gastrocnemius medialis muscle force-length characteristics. CONCLUSION: The prediction of muscle geometry and the prediction of force-length characteristics are very good with a simple model such as a planimetric model. This conclusion suggests that the influence of properties neglected in such a simple model are either small or are internally compensated for in the net effects. PMID- 11415796 TI - A method for developing biomechanical profiles of hand-intensive tasks. AB - OBJECTIVE: To develop a methodology for generic, comprehensive biomechanical profiling of hand-intensive tasks. DESIGN: Based on a multifactorial model of work-related musculoskeletal disorder development, a method was developed for continuous, simultaneous measurement of relevant variables (wrist and finger joint position, force, muscle activity, and carpal tunnel pressure). Joint dynamics and tendon travel were derived. BACKGROUND: Few generic dose-response relationships have been identified for work-related musculoskeletal disorders. This may improve if methodologies are developed that quantify multiple factors along several dimensions (means, cumulative exposure, etc.). This requires continuous, simultaneous measurements, and facilitates examination of interactions. METHODS: Five touch-typists were instrumented to quantify their biomechanical profiles using the methodology, and to evaluate the sensitivity of the method to various work organization/design conditions. RESULTS: The method captured individual and group responses to design conditions and revealed interactions and trade-offs between response variables. Carpal tunnel pressure was found to be sensitive to radial-ulnar wrist posture. CONCLUSIONS: Multi variable biomechanical profiling can provide insight into effects of work design on workers; however, to achieve statistical significance large numbers of subjects are needed. PMID- 11415797 TI - A biomechanical study of occupational loads in the shoulder and elbow in dentistry. AB - OBJECTIVE: The aim was to demonstrate that when electromyographic (EMG) recordings are supplemented by biomechanical calculations more detailed information on shoulder loads are produced. DESIGN: Shoulder and elbow moments were calculated for three work situations of arm abduction (> 30 degrees, < 30 degrees and < 30 degrees with force on the hand) and were related to EMG. BACKGROUND: Prolonged arm elevation may result in shoulder pain and information on shoulder loads is needed to avoid development of musculoskeletal disorders. METHODS: Three-dimensional coordinates from video recordings of eight working dentists were used in a static link segment model based on orthopaedic axes. Moments and EMG (trapezius muscle) were related. Results: Working with the upper arm abducted more than 30 degrees produced 1.7 Nm (440%) higher moments of shoulder abduction, 0.5 Nm (100%) higher moments of outward upper arm twisting and 0.3 Nm (18%) smaller moments of elbow flexion than working with the upper arm abducted less than 30 degrees. Trapezius muscle activity did not increase with increased arm abduction. Force on the hand (20 N) recorded during cavity filling altered moments of the shoulder and elbow but not the activity level of the trapezius muscle. CONCLUSION: Supplementing EMG with biomechanical calculations of moments results in more detailed information on shoulder loads. This is important for over-all risk identification. PMID- 11415798 TI - 3D shoulder position measurements using a six-degree-of-freedom electromagnetic tracking device. AB - OBJECTIVE: To describe a recording and processing methodology for obtaining kinematic data of the shoulder which meets three more criteria besides usual requirements regarding precision and accuracy: sufficient speed, obtaining complete 3D kinematics including joint rotations, and usage of coordinate systems based on reference points. DESIGN: Static recordings of shoulder bone orientations during standardized humerus elevations based on the palpation technique using a six-degree-of-freedom electromagnetic tracking device. BACKGROUND: An easy, fast, well standardized measurement methodology for obtaining complete 3D shoulder kinematic data is urgently needed for fundamental musculoskeletal and clinical research. METHODS: A measurement methodology was designed and developed. Shoulder kinematics were obtained from repeated measurements on 15 healthy subjects performed by two observers. Inter-trial, inter-day, inter-observer and inter-subject variability were established. Results were compared to literature. RESULTS: Complete kinematic descriptions were obtained. A measurement speed of about one position per second could be reached. The measured kinematics and accuracy of the measurements were found to be in concordance with the literature. CONCLUSION: All previously formulated criteria for a clinical useful method for obtaining shoulder kinematics have been met. PMID- 11415799 TI - Mechanical behavior of the female sacroiliac joint and influence of the anterior and posterior sacroiliac ligaments under sagittal loads. AB - OBJECTIVE: The purpose of this study was to examine the mechanical behaviour of the female sacroiliac joint and the effects of its two major ligaments to joint stability. DESIGN: A cadaveric model was used to study the mechanical behaviour of the sacroiliac joints, and sequential dissection was performed to examine the contribution of the anterior and posterior sacroiliac ligaments in joint stability. BACKGROUND: Instability of the sacroiliac joints have been suspected as a possible cause of low back pain. Despite several investigations on joint anatomy and joint mobility, its stabilising mechanism is still not clear. METHODS: Four fresh cadaveric specimens of the female pelvis were tested on an Instron material testing machine. Eccentric compressive force of 60% of the subject's body weight was applied to the pelvis through the sacrum. Relative three-dimensional six-degree-of-freedom movement at the left sacroiliac joints was recorded with a specially designed motion tracking device. The device has an accuracy of 0.01 mm and is compact enough to be mounted across the joint. The test was repeated after sequential selective dissection of the bilateral anterior, and then posterior sacroiliac ligaments. RESULTS: Rotation up to 1.2 degrees and translation up to 0.9 mm were measured from the intact specimens. Lateral rotation, which tended to open the top portion of the joint, and sacral nutation were the primary rotations. On average, the rotation angles increased 10% when either the anterior or posterior ligaments were cut, and 30% when both ligaments were cut. CONCLUSIONS: Lateral rotation and nutation rotation of the sacrum were found to be the predominant motion, though the values were limited to less than 1.2 degrees. Both the anterior and posterior sacroiliac ligaments were found to play an important role in resisting rotations at the joints. PMID- 11415800 TI - Age-related differences in body segmental movement during perturbed stance in humans. AB - OBJECTIVE: The temporal and spatial features of body segmental movements of the head, trunk, thigh, shank and foot in response to a fast speed postural disturbance were compared among four age groups. It was hypothesized that the upper body movement was minimized in the healthy young adults, but was augmented in the older adults. DESIGN: Repeated measures design with 12 experimental conditions. BACKGROUND: It is not well understood yet why the elderly fall more than the young. In studies of locomotion and active balance tasks, the head position has been suggested as a reference frame for controlling dynamic equilibrium. This study was aimed at examining: (1) whether or not this reference frame would exist during involuntary balance tasks, which are most likely related to accidental falls in the elderly; and (2) how would the stability of this reference frame change with age. METHODS: Thirty-six healthy subjects of four age groups (20-70 + yr of age) participated in this study. Postural perturbation was introduced by a moving platform. The sagittal plane kinematics of five body segments was directly measured using Integrated Kinematic Sensors. RESULTS: Compared to the lower limb movement, the upper body movement, especially the head, was minimal (less than 7 degrees ) in the young adults (age group 1), and was largest (about 14 degrees ) in the older subjects (age groups 2, 3 and 4). Moreover, the maximum range of motion of the older adults was found to be positively correlated not only to the height of the segment, but also to the timings of the movements, while the young adults showed no correlation between range of motion and the timings of the movement. CONCLUSIONS: The findings suggested that the young adults indeed stabilized their upper body as a reference frame for maintaining postural balance. In contrast, this reference frame was not stabilized well in the older adults. In compensation, the older adults adopted a more rigid movement strategy, compared to a more flexible movement strategy in the young. All these changes would lead the older subjects to be less tolerable to postural disturbances and more susceptible to falls than the young subjects. PMID- 11415801 TI - Positioning device for optimal active kinematic real-time magnetic resonance imaging of the knee joint: a technical note. AB - OBJECTIVE: A prototype of a positioning device created especially for the diagnostic imaging of the patellofemoral joint was developed in order to achieve reproducible examination conditions. DESIGN: For this purpose a clinical trial on healthy test persons was carried out under real examination conditions. BACKGROUND: A special real-time MRI technique (Local-Look-technique) makes the analysis of active functional motion images possible. A prerequisite for this technique is accurate reproducible positioning of the knee joint in the MRI unit. METHODS: This positioning device was evaluated during a total of 50 examinations of the knee joints of five healthy test persons. The right patellofemoral joint of each test person was examined twice at different time points in order to check the reproducibility of all examination conditions. Comparing two examination series from the same individual, reproducibility of MRI slices was guaranteed by using identical anatomical landmarks. Image quality and test-retest reliability were analyzed on the computer screen. RESULTS: Optimal desired fixation of the leg in the MRI tube using the device was accomplished in all cases. The extent of motion of the knee joints ranged from 38 degrees (n = 2) to 40 degrees (n = 3) of flexion to full extension, which was satisfactory for the evaluation of the patellofemoral joint. Free movement of the patella and the lower leg was observed. The active functional MRI examination using this device was satisfactory and reproducible as assessed by test-retest reliability. CONCLUSIONS: The positioning device is a useful development in achieving active functional real-time MRI evaluation of the knee and patellofemoral joint. Using this diagnostic tool, reproducible quantitative examinations can now be easily performed. The reproducibility and high reliability as well as its simplicity of operation render this diagnostic tool suitable for use in orthopedics and traumatology. PMID- 11415802 TI - Quadriceps inhibition following arthroscopy in patients with anterior knee pain. AB - OBJECTIVES: This study was aimed at investigating muscle strength and quadriceps inhibition in patients with unilateral anterior knee pain syndrome. DESIGN: Functional assessments were made before an arthroscopic knee surgery and 6 weeks and 6 months following the surgical intervention. BACKGROUND: Traumatic knee injuries have been associated with severe muscle inhibition of the knee extensor muscles. Muscle inhibition is a serious hindrance in the rehabilitation process and prevents full functional recovery of the affected joint. METHODS: 30 patients participated in the study. Isometric quadriceps strength was measured using a KinCom dynamometer. Muscle inhibition was assessed using the interpolated twitch technique which requires applying a single electrical twitch to the femoral nerve during a maximal isometric knee extensor contraction. Pain was assessed with a 100 mm visual analogue pain scale. RESULTS: Pre-surgery, substantial muscle inhibition and pain was observed in the affected leg compared with the contralateral leg. Over the 6 month period there was a decrease in pain and muscle inhibition, although the decrease in muscle inhibition failed to reach statistical significance. Muscle strength showed a decrease 6 weeks post-surgery followed by an increase 6 months following surgery compared with pre-surgical values. CONCLUSIONS: The arthroscopic intervention was successful in reducing pain in patients with unilateral anterior knee pain syndrome. However, muscle inhibition was still substantial 6 months following surgery and was significantly higher in the affected and contralateral limb than in normal subjects. PMID- 11415803 TI - A new method for evaluating motor control in gait under real-life environmental conditions. Part 1: The instrument. AB - OBJECTIVE: To devise an instrument to measure linear acceleration of the trunk during gait under real-life environmental conditions. DESIGN: A mathematical algorithm is suggested to transform data to a horizontal-vertical coordinate system. Calibration procedures are described. BACKGROUND: A portable, low mass device to measure linear acceleration of the trunk in a horizontal-vertical coordinate system during prolonged walking is of clinical interest and has not been described previously. METHOD: Linear acceleration is registered by a triaxial, piezoresitant accelerometer, sampled at 512 Hz and collected by exchangeable 20 Mb memory cards of a body-mounted data-logger. Data are transformed to a horizontal-vertical coordinate system, utilising the accelerometer's capacity as an inclinometer. Calibration testing was done in a servohydraulic jig by measuring acceleration of 5 Hz, 16 mm vertical displacements, first along each of the sensing axis, then by a tilted accelerometer. RESULTS: Precision testing of each axis and transformed data from all axes showed low variability for acceleration root mean square of sequences of 1000 samples (CV <0.001). Accuracy testing by double integration of transformed data from the tilted accelerometer demonstrated a mean absolute difference of 0.04 mm (SD = 0.03 mm) from the programmed input. CONCLUSIONS: Data from the arbitrarily tilted accelerometer transformed to a horizontal-vertical coordinate system gave precise and accurate measurements of linear accelerations when tested in a servohydraulic jig. Procedures for applying the device in gait analysis are described in a companion article. RELEVANCE: Control of upper body and body centre of mass are important aspects of balance, but gait analysis has largely focused on the movements of the lower limbs. This device can be used to register acceleration of a reference point on the upper body during walking, and may thus give new information about balance in gait. PMID- 11415804 TI - A new method for evaluating motor control in gait under real-life environmental conditions. Part 2: Gait analysis. AB - OBJECTIVE: The validity of assessing balance in gait by measuring balance in standing is questionable. Better methods for measuring balance during walking are therefore needed. DESIGN: It is suggested that the individual will demonstrate adequate postural control by moving a reference point near the body centre of mass (CoM) smoothly towards an intentional goal, even though movements of the extremities show variability consistent with a changing environment. BACKGROUND: In spite of an increased interest in variability as a prerequisite for motor control, gait analysis methods focus, to a large extent, on symmetry and repeatability of movements in stereotyped settings. METHODS: Acceleration of a reference point over the lumbar spine is registered during walking by a portable, triaxial accelerometry system. RESULTS: A quadratic relation between acceleration root mean square (RMS) and walking speed is demonstrated, and a second degree polynomial can therefore be computed as a curve estimate, if acceleration RMS representing at least three walking speeds are available. CONCLUSIONS: The relation between acceleration over a reference point on the trunk and walking speed can be compared between trials and also when walking speeds are self selected. Calibration procedures and testing of the instrument for precision and accuracy in a mechanical testing jig are described in a companion article. RELEVANCE: This study suggests a new alternative to the traditions of measuring balance in standing and movements of the legs in walking. The method allows balance in gait to be assessed at self-selected speeds in relevant environmental conditions, which may facilitate gait analysis in the clinic and improve the validity of the results. PMID- 11415805 TI - Fatigue during repeated eccentric-concentric and pure concentric muscle actions of the plantar flexors. AB - OBJECTIVE: To compare the fatigue development during pure concentric actions with eccentric-concentric actions in standardized isokinetic, open-chain conditions. DESIGN: The study was performed with measurement of muscular strength and EMG of the plantar flexors on an isokinetic dynamometer. BACKGROUND: Fatigue development on a dynamometer has mostly been studied with repeated pure concentric actions. Eccentric-concentric actions are, however, often used in normal activities. An eccentric action has an immediate effect on a following concentric action and therefore it is of interest to follow the fatigue process in repeated combined actions. METHODS: Ten healthy young women participated. The pure concentric test was performed with the same number of cycles as in the eccentric-concentric test, which was performed until exhaustion. Development of MPF and RMS of the EMG in the triceps surae, as well as work, were evaluated. RESULTS: The total amount of work was significantly higher in the eccentric-concentric test than in the pure concentric test. The work decreased similarly (32-39%) with no significant difference between the two tests. A significant decrease (12-16%) in MPF was only seen in the eccentric-concentric test. No changes in RMS were seen. CONCLUSION: The similar reductions of work seen in both tests should primarily be sought in the contractile process as such. Changes in MPF may then be viewed as a phenomenon dependent on the timing of the duty cycle. RELEVANCE: The eccentric concentric test lead to exhaustion and showed EMG changes and should therefore be recommended when evaluating the development of fatigue in muscular performances in different groups of patients. PMID- 11415806 TI - Torque, work and EMG development in a heel-rise test. AB - OBJECTIVE: To study the fatigue process in the calf muscle during a standing heel rise test. DESIGN: Heel-rises were performed on a force plate measuring development of torque in the ankle joint. BACKGROUND: The heel-rise test is a commonly employed clinical test to evaluate the function of the calf muscle by counting the number of heel-rises before exhaustion. Development of fatigue during other eccentric-concentric actions has earlier been studied as decreases in force, work and EMG. METHODS: Ten healthy males (mean age 25 yr) participated in the study. Torque and work were calculated using measurements from a force plate. Mean power frequency and root-mean-square of the EMG in the triceps surae were evaluated separately in the eccentric and concentric phases. RESULTS: Increases of mean torque during both the eccentric and concentric phases were found. Work performed decreased during the concentric phases due to decreased range of motion. No changes in root-mean-square and decreases in mean power frequency during the concentric phases indicated development of muscular fatigue, whereas decreases in both root-mean-square and mean power frequency during the eccentric phases indicated decreasing muscular activity. CONCLUSIONS: Accordingly, the limiting factor for the heel-rise test was not loss of muscle force at the range of motion used, but rather a failure to maintain the initial range of motion owing to muscle fatigue. RELEVANCE: This method of calculating torque development in the ankle joint provides an opportunity to study the fatigue process in terms of performance. The results show that the heel-rise test reflects muscle endurance rather than strength of the calf muscle. PMID- 11415807 TI - Mechanical properties of the normal human tibial cartilage-bone complex in relation to age. AB - OBJECTIVE: This study investigates the age-related variations in the mechanical properties of the normal human tibial cartilage-bone complex and the relationships between cartilage and bone. DESIGN: A novel technique was applied to assess the mechanical properties of the cartilage and bone by means of testing the cartilage-bone complex. BACKGROUND: Up to now, mechanical testing of cartilage and bone has been reported separately, and little is known about the mechanical behaviour of both tissues when examined as a unit. METHODS: Cylindrical human proximal tibial cartilage-bone complex specimens from 31 normal donors aged 16-83 years were tested in compression. The deformation was measured simultaneously in bone and cartilage to obtain the mechanical properties of both tissues. RESULTS: The stiffnesses and elastic energies of both cartilage and bone showed an initial increase, with maxima at 40 years, followed by a steady decline. The viscoelastic energy was maximal at younger ages (16-29 years), followed by a steady decline. The energy absorption capacity did not vary with age. Stiffnesses and elastic energies were correlated significantly between cartilage and bone. CONCLUSIONS: The present study demonstrates that similar age related trends were seen in cartilage and bone, as if they behaved as a single mechanical unit. RELEVANCE: The basic information presented here on the mechanical properties of cartilage and bone and the correlations between them reveals the unit function of both tissues that are of importance for the understanding of the etiology and pathogenesis of degenerative joint diseases, such as arthrosis. PMID- 11415808 TI - The effects of external mechanical stimulation on the healing of diaphyseal osteotomies fixed by flexible external fixation. AB - OBJECTIVE: The purpose of this study was to investigate the effect of an externally applied mechanical stimulus on fracture healing under flexible fixation. DESIGN: Stimulation of fracture healing under various conditions of interfragmentary movement in an in vivo fracture model on 41 sheep. BACKGROUND: It is generally accepted that small interfragmentary movements (IFMs) yield better bone healing results than larger IFMs (> 1 mm). However, the optimal size of IFM within the 1-mm range remains undetermined. METHODS: Standardized transverse osteotomy of 3 mm gap size in the left ovine tibia was fixed with an unilateral external fixator. The sheep were divided into four IFM groups of 0.0, 0.2, 0.4 and 0.8 mm and stimulated with this amplitude for 1200 cycles per day at 1 Hz. After a healing period of 6 weeks, bone mineral density and biomechanical stability were evaluated to determine the quality of healing. RESULTS: The amount of callus formation increased significantly with increasing IFM (P <0.05). However, highest biomechanical stability of the healed bone and mineral density of the gap tissue was achieved with an IFM of 0.4 mm, although the differences were not significant. CONCLUSIONS: These results suggest that the optimal interfragmentary movement for acceleration of delayed fracture healing is in the range of 0.5 mm. However, the enhancement of the healing of flexibly-fixed fractures by external application of interfragmentary movement is limited. RELEVANCE: In this model the external application of a mechanical stimulus in addition to the stimulation caused by normal loading and the flexibility of the fixation did not enhance the healing process significantly. It appears that the external application of interfragmentary motion is promising perhaps only for patients unable to stimulate their fracture healing by weight-bearing. PMID- 11415809 TI - Measurement of surface contact area of the ankle joint. AB - OBJECTIVE: To determine the distribution of contact area of the ankle joint with axial loading and in positions of maximal dorsiflexion, plantar flexion, supination, and pronation. We also tested the effects of extrinsic tendon loading and arch instability. DESIGN: Nine cadaveric feet were studied in the intact condition and following transection of ligaments to create arch instability. BACKGROUND: Assessment of ankle contact in various joint positions and degrees of instability is difficult to accomplish with conventional methods. METHODS: Displacement of the talus relative to the tibia was measured with a magnetic tracking device and tibiotalar joint contact from proximity calculations of digitized joint surfaces. RESULTS: Contact area did not change significantly from unloaded condition to 667 N load condition in the medial, central, and lateral zones. Central zone contact area decreased in plantar flexion by an average of 324 mm(2) (SD, 165 mm(2)) (P = 0.0004). Medial zone contact area decreased in plantar flexion by a mean of 55 mm(2) (SD, 28 mm(2)) (P = 0.0004), decreased in pronation by an average of 42 mm(2) (SD, 36 mm(2)) (P = 0.0086), and increased in supination by an average of 20 mm(2) (SD, 26 mm(2)) (P = 0.0430). Lateral zone contact decreased in plantar flexion by a mean of 124 mm(2) (SD, 57 mm(2)) (P = 0.0002). CONCLUSIONS: In plantar flexion, there was a decrease in contact area. Loading extrinsic tendons to the foot did not significantly increase ankle contact area, but arch instability caused a decrease in central and lateral zone contact area. RELEVANCE: This technique was used to assess joint contact characteristics in various loading conditions and will be useful for evaluating the extent to which treatment for ankle or hindfoot problems such as bracing or reconstruction operations restores normal joint contact. PMID- 11415810 TI - Quantifying lateral pelvic displacement during walking. AB - OBJECTIVE: The purpose of this investigation was to test a new procedure for quantifying lateral pelvic displacement during walking. DESIGN: A quasi experimental design was used to quantify the gait of 18 unimpaired people and one person with hemiplegia. BACKGROUND: Although previous techniques provided useful information on amplitude of lateral pelvic displacement, they did not consider step-to-step variations in walking direction or enable quantification of symmetry. METHODS: Three-dimensional motion analysis was used to collect the coordinates of light-reflective markers placed on the scarum and heels of each subject. Subjects performed one 10 m overground walk at their preferred speed. Amplitude and symmetry of lateral pelvic displacement were quantified relative to the step-to-step variation in the path of motion (base of support). RESULTS: The mean amplitude of lateral pelvic displacement for the unimpaired group was 40.8 mm, and symmetry was 3.1 mm. The amplitude of lateral pelvic displacement for the hemiplegic person was 88.4 mm. Symmetry was 30.9 mm, with deviation toward the non-paretic side. CONCLUSION: The new procedure provided information on the amplitude and symmetry of lateral pelvic displacement in unimpaired adults and was sensitive to deviations of a person with a walking abnormality. RELEVANCE: Treatment of atypical lateral pelvic displacement is frequently an aim of stroke rehabilitation. Therefore, it is important to have objective, accurate methods of quantification. PMID- 11415811 TI - Reliability of weight-bearing heel pad thickness measurements by ultrasound. AB - OBJECTIVE: To evaluate the intra- and inter-operator reliability of measuring weightbearing heel pad thickness in healthy subjects. BACKGROUND: Heel pad thickness has been reported to be an important factor in determining stresses observed in healthy as well as pathological feet. To date no published data are available on inter- and intra-operator variation in sonographic heel pad thickness measurements in adults. METHOD: bilateral heel pad thickness was measured in 10 adult subjects, with no history of plantar heel pain, by two experienced operators, on two separate occasions. Limits of agreement for three replicate measurements by each ultrasonographer and for replicate measurements were determined. RESULTS: Replicate heel pad thickness measurements in 95% of cases differ by less than 0.39 mm, and the magnitude of variations is similar when measurements are made by either single or different ultrasonographers, and were similar for right and left heel pad thickness measurements. Between scan measurements demonstrated a coefficient of variation of 3.2%. CONCLUSION: Weightbearing heel pad thickness measurements by ultrasound using a standardised technique in normal adult feet are reasonably reliable. RELEVANCE: Heel pad thickness can be measured reliably. Heel pad thickness may be an important predictor in the cause of ulceration in diabetics, and a potential risk factor in plantar pain affecting athletes. Further work assessing the heel pad in pathological conditions and athletes merits further study. PMID- 11415812 TI - The morphology and biomechanics of latissimus dorsi. AB - OBJECTIVE: To determine the morphology of the latissimus dorsi in order to assess its actions on the shoulder, the lumbar spine and the sacroiliac joint. DESIGN: A dissection study accompanied by an analysis of the force vectors of the muscle and its parts. BACKGROUND: Although recognised as a muscle of the shoulder, latissimus dorsi has been accorded a role as an extensor of the lumbar spine, and is said to brace the sacroiliac joint. Consideration of the anatomy of the latissimus dorsi suggests that the magnitude of these actions has been overstated. METHODS: The fascicular anatomy of the latissimus dorsi was determined by dissection in five adult cadavers. The size, attachments, and orientation of each fascicle were determined. By applying a force coefficient the maximum force of each fascicle was estimated from its physiological cross sectional area. By summing the forces and moments of each fascicle the maximum force exerted by latissimus dorsi was calculated for its actions on the shoulder, the lumbar spine, and the sacroiliac joint. RESULTS: The latissimus dorsi was found to consist of a series of fascicles with segmental attachments to the lower six thoracic spinous processes, the L1 and L2 spinous processes, the lateral raphe of the thoracolumbar fascia, the iliac crest and the lower three ribs. These fascicles were uniform in size across a given muscle but varied from specimen to specimen. The maximum total force exerted by the latissimus dorsi on the shoulder was estimated to range between 162 and 529 N, but in view of the attachments of the muscle, only a portion of that force can be exerted on the lumbar spine. The maximum extensor moment exerted on the lumbar spine was calculated to be 6.3 N m. The maximum force exerted across the sacroiliac joint was calculated to be 30 N. CONCLUSIONS: The latissimus dorsi is designed to move the upper limb or to raise the entire trunk in brachiation. Its possible contribution to extension of the lumbar spine is trivial as is its capacity to brace the sacroiliac joint. RELEVANCE: Despite assertions and concerns to the contrary, the latissimus dorsi is of little mechanical importance in the lumbosacral region. PMID- 11415813 TI - Three dimensional moments in the lumbar spine during asymmetric lifting. AB - OBJECTIVE: To better understand the loads placed on the spine during asymmetric lifting. DESIGN: Analysis of variance was used to test the effects of asymmetry (0, 45, 90 degrees ), event (up, down, min and max), and trial (first and second) on applied spine moments and resulting EMG signals in the lumbar spine. BACKGROUND: Loading conditions resulting from symmetric lifting are well documented in the literature, yet free-style asymmetric tasks have not been intensively studied. METHODS: Infrared markers and EMG electrodes were fixed to ten subjects. Subjects were asked to lift a 45 N weight from three different positions in relation to the sagittal plane. External loads on the L3-L4 and L4 L5 disc planes were calculated. Maximum moments and EMG were compared with those at lift off and set down of the burden. All lifts were repeated twice. RESULTS: Introducing asymmetry to the lift increased lateral bending and twisting moments. Flexion moments increased by approximately 5% after lift off of the load. During the second repetition of the lift, flexion and twisting moments decreased and increased, respectively. EMG signals were greater on the contralateral side during asymmetric lifts. CONCLUSIONS: Asymmetric lifting places complex loads across the trunk, which may become increasingly asymmetric with repeated trials. RELEVANCE: Individuals should take precautions to perform lifts in a symmetric manner and to avoid repetitive asymmetric lifting if possible. Past study indicated that twisting and lateral bending loads, when coupled with flexion loads, resulted in much greater disc compression and should be avoided. This work described the degree to which lateral bending and twisting loads were introduced as lifts deviated from the sagittal plane. PMID- 11415814 TI - In vivo determination of contact areas and pressure of the femorotibial joint using non-linear finite element analysis. AB - OBJECTIVES: A three dimensional finite element model of the femorotibial joint was developed from MR images in order to quantify in vivo the articular contact. BACKGROUND: Most of femorotibial joint models were elaborated from in vitro experiments. The stereophotogrammetric technique was used to model the geometry and mechanical testing had been performed to quantify the material properties. METHOD: MR images were performed on a normal adult knee joint, in extension position. An image processing software developed in our laboratory allowed our model geometry to be constructed, and a pre-and post-processing software allowed us to develop a three-dimensional finite element model. Experimental contact area values were obtained using a method developed in our laboratory. Theoretical contact values, areas and hydrostatic pressure were obtained with a non-linear finite element computation using a non-linear software solver. RESULTS: The results show a good agreement between theoretical and experimental contact area values. Hydrostatic pressure was found to be higher at the medial contact than at the lateral contact. CONCLUSION: This study validated the use of contact elements to quantify the contact areas. The model permitted the body weight simulation to understand the role of the menisci. RELEVANCE: The clinical application of the study was to develop a method evaluating the influence of rotational abnormalities of the lower limbs on the knee joint at short- and long-term. This consisted of quantifying the contact area and pressure values and their migration. PMID- 11415815 TI - Determinants of cruciate-ligament loading during rehabilitation exercise. AB - OBJECTIVES: To predict and explain the pattern of cruciate-ligament loading during squatting exercises; to determine the effect of hamstrings co-contraction on anterior cruciate ligament (ACL) load during squatting; and to determine the effect of the weightbearing force on ACL load during squatting. DESIGN: Mathematical modeling of the human musculoskeletal system. BACKGROUND: Squatting is a commonly prescribed exercise for strengthening the muscles of the thigh following ACL reconstruction. Although the forces induced in the ACL are purported to be low, no experimental data are available to corroborate this claim. The reason is that measurements of knee-ligament forces are difficult to obtain in vivo. METHODS: The human body was modeled as a four-segment, six degrees of freedom, planar linkage. The hip, ankle and toes were each modeled as a hinge joint. The relative displacements of the femur, tibia and patella were calculated using a three-degrees of freedom, sagittal-plane model of the knee. Eleven elastic were used to describe the geometric and mechanical properties of the knee ligaments. The model was actuated by 22 musculotendinous units. Optimization theory was used to calculate the forces developed in the muscles and the forces transmitted to the knee ligaments during squatting. RESULTS: The model ACL was loaded from full extension to 10 degrees of knee flexion during squatting; the model PCL was loaded at knee-flexion angles greater than 10 degrees. The pattern of cruciate-ligament loading is determined by the shapes of the articulating surfaces of the bones and by the changing orientation of the hamstrings muscles at the knee. Hamstrings co-contraction is the major determinant of ACL loading during squatting exercises; the weightbearing force has a relatively small effect on the force induced in the ACL. CONCLUSION: The calculations support the contention that squatting is a relatively safe exercise for strengthening the muscles of the thigh following reconstruction of the ACL. RELEVANCE: Knowledge of the forces borne by the knee ligaments is important for designing exercise regimens subsequent to ligament injury and repair. The quadriceps and hamstrings muscles may be strengthened without loading a newly reconstructed ACL by performing squats with the knee flexed to 10 degrees and greater. PMID- 11415816 TI - Kinematics of valgus bracing for medial gonarthrosis: technical report. AB - OBJECTIVE: To determine whether a valgus knee brace altered the alignment of the shank segment with respect to the thigh segment of patients with medial gonarthrosis of the knee. DESIGN: Twelve (subject) x 2 (brace condition) x 2 (speed) repeated measures design. METHODS: Twelve male subjects (mean age 55 yr, SD 5 yr) who were prescribed a valgus knee brace for medial gonarthrosis walked on a treadmill at normal (mean 1.5 m/s, SD 0.3 m/s) and fast walking speed (mean 2.1 m/s, SD 0.2 m/s) while filmed by two gen-locked video cameras. Flexion, coronal and axial rotation angles of the thigh and shank segments in a global co ordinate system were determined from kinematic data of reflective markers on the affected leg. Data were calibrated with respect to the standing 'no-brace' condition and averaged over five step cycles at four specific points in time during the stance phase. RESULTS: The brace had no effect on thigh coronal angle but reduced shank coronal angle at toe-off (statistically significant). The brace also prevented full extension (statistically significant) during mid-stance by resisting valgus forces in the coronal plane transmitted though the brace's helical strap. This same strap was believed to be responsible for axial forces that created statistically significant external rotation of the thigh axial angle throughout the stance phase and internal rotation of the shank axial angle during knee extension at heel strike and mid-stance. The brace hinge may explain the unexpected external rotation of the shank axial angle that accompanied knee flexion. CONCLUSION: This study demonstrated that the brace altered the kinematics in all three planes of motion and transmitted forces from one plane to another. This study also indicates that the subject-brace interaction was more complex in nature than initially anticipated. RELEVANCE: Clinical evidence suggests that there can be a reduction of knee pain by wearing a valgus brace for medial gonarthrosis and that the brace alters the orientation of thigh and shank segments during gait in such a manner to reduce the inter-joint pressure of the afflicted compartment. Therefore a biomechanical analysis of a possible kinematic effect was warranted. PMID- 11415817 TI - A dynamic cadaver model of the stance phase of gait: performance characteristics and kinetic validation. AB - OBJECTIVE: This study was undertaken to evaluate the performance of a new dynamic laboratory model of the stance phase of gait. DESIGN: Five cadaver feet were repetitively tested in the apparatus. BACKGROUND: Typical biomechanical investigations of cadaver feet simply place a static load on the tibia. The present system was designed to better simulate the changing in-vivo loading environment of the foot and ankle during gait. METHODS: The device mimics the behavior of the tibia, foot, and ankle from heel-strike to toe-off by reproducing the physiologic actions of five extrinsic foot muscles and physiologic motion at the proximal tibia. To verify its utility, cadaver gait simulations were conducted while measuring applied muscle forces, ground reaction forces, and plantar pressures. RESULTS: Dynamic muscle forces were consistently delivered to within 10% of pre-programmed values. Dynamic measurements of ground reaction forces and plantar pressure were similar to those measured in healthy human subjects. Peak vertical (y), foreaft (x) and medio-lateral (z) forces were 110, 18, and 4% of body weight respectively. Compressive force in the tibial shaft reached 410% of body weight. RELEVANCE: Cadaver studies have greatly enhanced our understanding of normal and pathologic foot function, but are often limited by over-simplified loading conditions. The apparatus presented here accurately reproduces the in-vivo loading environment and provides a powerful investigational tool for the study of foot and ankle function. With this device, musculoskeletal structures can be examined in detail under biomechanical conditions similar to those they experience in life. PMID- 11415818 TI - A kinematic comparison of overground and treadmill walking. AB - OBJECTIVE: To compare overground and treadmill ambulation for possible differences in gait temporal variables and leg joint kinematics. DESIGN: A human subject trial of walking in two conditions. BACKGROUND: The treadmill is frequently used to simulate overground ambulation; however, the literature shows a wide difference of opinion as to whether the treadmill replicates the overground environment. METHODS: A total of 17 uninjured subjects walked overground at their preferred velocity. The treadmill was then set at the average velocity obtained in overground walking. Gait temporal variables and leg joint kinematics were analysed using the three dimensional (3D) Kinemetrix Motion Analysis System. The data were analysed separately for the two gender groups and for the groups combined. RESULTS: In the females, only the maximum hip flexion angle was significantly different in the two conditions with greater flexion occurring on the treadmill. For males, significant differences were noted between the two conditions for cadence and maximum knee flexion angle with greater values in the treadmill walking. When all subjects were compared, significant increases were seen during treadmill walking in hip range of motion, maximum hip flexion joint angle and cadence, while a significant decrease was observed in stance time. CONCLUSIONS: Statistically significant differences exist between overground and treadmill walking in healthy subjects for some joint kinematic and temporal variables. RELEVANCE: The results of this and other studies demonstrate that the kinematics of human walking differ in treadmill and overground ambulation. The next step in this line of research is to assess whether the kinematic differences in the two walking conditions are exhibited in different patient groups. If differences do exist in patient groups, then the interpretation of treadmill based gait analysis findings should not be extended to that of ground walking, particularly where the patient has an impaired hip function, such as that exemplified after total hip replacement. PMID- 11415819 TI - Stability of external circular fixation: a multi-variable biomechanical analysis. AB - OBJECTIVE: To determine how the manipulation of the parameters of fixation and components of the circular external frame could improve and maintain optimal stability of bone fragments. DESIGN: We performed a multi-parametric biomechanical analysis of the extrinsic parameters effecting bone fragment stabilization. Results of testing are presented as a percent change in stiffness due to the manipulation of frame components and their interaction with other fixation parameters. BACKGROUND: Although there have been investigations of the biomechanical characteristics of circular external fixation, they have been limited to either individual frame components or full frame comparisons. Therefore, these studies did not provide a comprehensive understanding of how the manipulation of circular fixator components influences bone fragment stability. METHODS: Mechanical testing was performed in three phases examining the effect of numerous components including ring diameter, wire angle, ring separation, etc. on axial, torsional and bending stiffness. RESULTS: For phase I (single ring) and phase II (double-ring block), ring diameter was the most significant factor affecting axial and torsional stiffness, while wire angle, ring separation, and their interaction had the most influence on bending stiffness. Phase III (two double-ring blocks) showed that ring positioning with respect to the osteotomy site had the most affect on bending and torsional stiffness while axial stiffness was non-linear and dependent upon the applied load. CONCLUSIONS: The stability of bone fragments within a circular external fixator is affected by manipulation of the parameters of fixation or individual components of the frame. The contribution of each component to overall bone fragment stability is dependent upon the mode of loading. The changes in overall stability of bone fragments are dependent not only on the individual frame components but also upon their interaction with other parameters of fixation. RELEVANCE: Understanding how the manipulation of individual frame components will affect overall bone fragment stabilization will allow the surgeon to better control the stability of bone fragments for each clinical situation. PMID- 11415820 TI - Twist knot cerclage wire: the appropriate wire tension for knot construction and fracture stability. AB - OBJECTIVE: The aim was to find the best wire tension in order to permit a reliable first twist and simultaneously provide the best stability of fracture fixation from the twist knot cerclage wire. DESIGN: Wires at different distal tensions, looped around the fracture, were measured during twist and compared with the yield strength of the wire. Then, the fracture stability of the twist knot cerclage wire was determined from the pull-out strength. METHODS: In order to measure wire tension during twist knot construction, an instrument was designed using the tension load cell of a universal testing machine, a 15 degrees oblique osteotomy femoral shaft and 1.25 mm diameter wire. A wire tensioner and a pair of extraction grips were then used for measuring the pull-out strength of the cerclage wire fixation. RESULT: Three wire tensions (160, 200 and 240 N) were used as looped wire for the first twist knot construction. The 200 N tension cerclage wire provided the best fracture stability. CONCLUSION: It was found that 200 N was the best wire tension for the construction of a twist knot cerclage wire. RELEVANCE: When a cerclage wire is twisted at a femoral shaft using 1.25 mm diameter wire, a wire tension of 200 N should be used to achieve a reliable first twist and the best stability of fracture fixation. PMID- 11415821 TI - Some effects of camera placement on the accuracy of the Kinemetrix three dimensional motion analysis system. AB - OBJECTIVE: To assess the accuracy of the Kinemetrix motion analysis system to measure horizontal movement by a single reflective marker away from two cameras with differing camera placements. DESIGN: Trial of the effects of nine different camera arrangements on precision of non-human movement. BACKGROUND: In many cases the ability to arrange cameras to allow a separation of 60 degrees is not possible. Little is known about the precision of motion analysis systems for small camera angle separations. METHODS: The accuracy of the Kinemetrix system was assessed with camera horizontal separations of 15 degrees, 30 degrees and 45 degrees, and vertical separations of 0 degrees, 15 degrees and 30 degrees rendering nine different camera placements. The distance between the cameras and the object was always maintained at 4 m. During each test the marker was moved a known horizontal distance along a line bisecting the horizontal angular separation of the two cameras. The mean absolute errors of the Kinemetrix measurement were calculated. RESULTS: At the smallest camera separation tested (15 degrees horizontal, 0 degrees vertical), the Kinemetrix was unable to calculate the three-dimensional co-ordinate of the marker. For all other camera positions tested, the errors in measurements were small (mean absolute errors < 2 mm). CONCLUSIONS: Maintaining camera horizontal and vertical separations above a sum of 30 degrees is sufficient for clinical testing. RELEVANCE: Motion analysis systems are becoming more common for clinical evaluation where only confined testing areas are available. These confined areas often make positioning of cameras at greater than 60 degrees impossible; therefore, there is a need to explore the errors involved in placing two cameras at less than 60 degrees. PMID- 11415822 TI - Three-dimensional determination of femoral-tibial contact positions under in vivo conditions using fluoroscopy. AB - OBJECTIVE: A method has been developed to accurately measure three-dimensional (3 D) femoral-tibial contact positions of artificial knee implants in vivo from X ray fluoroscopy images using interactive 3-D computer vision algorithms. DESIGN: A computerized graphical (CAD) model of an implant component is displayed as an overlay on the original X-ray image. An image matching algorithm matches the silhouette of the implant component against a library of images, in order to estimate the position and orientation (pose) of the component. The operator further adjusts the pose of the graphical model to improve the accuracy of the match. BACKGROUND: Previous methods for in vivo measurement of joint kinematics make only indirect measurements of joint kinematics, require invasive procedures such as markers or pins, or make simplifying assumptions about imaging geometry which can reduce the accuracy of the resulting measurements. METHODS: Fluoroscopic videos are taken of implanted knees in subjects performing weight bearing motion. Images from the videos are digitized and stored on a computer workstation. Using computerized model matching, the relative pose of the two knee implant components can be determined in each image. The resulting information can be used to determine where the two components are contacting, the area of the contact region, liftoff angle, and other kinematic data. RESULTS: Accuracy tests done on simulated imagery and in vitro real imagery show that the pose estimation method is accurate to less than 0.5 mm of error (RMS) for translations parallel to the image plane. Orientation error is less than or equal to 0.35 degrees about any axis. Errors are larger for translations perpendicular to the image plane (up to 2.25 mm). In a clinical study, the method was used to measure in vivo contact points, and characterize the kinematic patterns of two different knee implant designs. CONCLUSIONS: The ability to accurately measure knee kinematics in vivo is critical for the understanding of the behavior of knee implant designs and the ultimate development of new, longer lasting implants. RELEVANCE: This work shows that it is possible to accurately measure the three-dimensional position and orientation (pose) of artificial knee implants in vivo from X-ray fluoroscopy images using interactive 3-D computer graphics. The method can be applied to any joint when accurate CAD models are available. The resulting data can be used to characterize the kinematics of current knee implant designs. PMID- 11415823 TI - In vivo determination of homogenised mechanical characteristics of human tibia: application to the study of tibial torsion in vivo. AB - OBJECTIVE: The study presents a method allowing the in vivo homogenised characteristics of the tibiae of children to be assessed. DESIGN: Studies have been performed on two groups of children: six normal children, aged from 5 to 16 yr, and on four children, aged from 8 to 11 yr with tibial deformities. We analysed the tibial transverse sections from CT scans performed on the left tibia of each child. BACKGROUND: Most tibial torsion studies have only been based on geometrical parameters. Our study integrated mechanical and geometrical considerations. METHODS: The finite element models and integration of mechanical properties were performed from CT scans. Then homogenised mechanical characteristics (tensile stiffness, flexural stiffness and torsional stiffness) were calculated. RESULTS: The homogenised mechanical characteristics decrease between 20 to 80% of the tibial length. The values increased with age for both groups of children. Children with abnormalities seem to have values of tibial rigidities comparable with those of normal tibiae. CONCLUSIONS: By considering the mechanical and geometrical properties of the tibia in our study, we showed that the bone stiffness of children is not altered with torsional deformities. RELEVANCE: Torsional tibial abnormalities of children are a frequent phenomenon which may have important consequences on gait and joints. The method developed could be used as an objective assessment of bone rigidities for analysing tibial disorders such as torsional abnormalities of varying severity. PMID- 11415824 TI - Effect of ovariectomy and calcium deficiency on the ultrasound velocity, mineral density and strength in the rat femur. AB - OBJECTIVE: Ultrasound transmission velocity (UTV) and bone mineral density (BMD) were examined as possible predictors of the bending strength of osteoporotic long bones in ovariectomized rats treated with calcium-depleted diet. BACKGROUND: Changes in cancellous bone quality due to osteoporosis have been well documented, but only a few studies described non-invasive assessment of the biomechanical quality of long bones. METHODS: Forty-three rats were divided into three groups: ovariectomized and calcium-depleted diet, ovariectomized and standard diet, and untreated controls. All femora were scanned by quantitative computed tomography (QCT) to measure BMD and cortical dimensions at the mid-diaphysis. Bending strength was determined from the maximum bending moment and the femoral cortical dimensions at the mid-diaphysis. The UTV of cortical bond at the mid-disphysis of the femur was measured in the proximal/distal direction. RESULTS: It was found that bending strength was significantly different among the groups (p<0.0001). Despite the fact that bending strength of the osteoporotic rat femur significantly decreased, the osteoporotic effect did not produce a significant change in cortical thickness. UTV in the ovariectomized and calcium-depleted diet was lower than in the other groups; however, no significant differences were found betwene those on the standard diet and the untreated controls. The BMD in ovariectomized rats was also significantly lower than in the untreated controls (p<0.05); however, the BMD in those fed with the calcium-depleted diet was not significantly different from those with the standard diet. Both cortical UTV and BMD as a single variable may not predict changes in bending strength of the cortical bone at the mid-diaphysis of the rat femur. However, the combined effect of UTV and BMD was significantly better than a single parameter in the assessment of strength. RELEVANCE: Osteoporotic fracture results from the reduced BMD and the microstructural changes involving the entire skeleton. The ovariectomized rats with calcium-depleted diets may simulate a model for the human postmenopausal osteoporosis. Ultrasound measurements of cortical bone may not be better predictors of osteoporotic fracture risk than those of cancellous bone. The strength of osteoporotic long bone may not be properly predicted by either BMD or UTV as a single determinant, measured at the cortical bone of the rat femur. PMID- 11415825 TI - The effect of fracture malunion at the mid-shaft of the metacarpal on the extrinsic muscle forces. AB - OBJECTIVE: To investigate the combined effect of dorsal angulation and shortening in metacarpal mid-shaft fracture malunions on the extrinsic flexion and extension muscle forces. DESIGN: An experimental study on human cadavers. BACKGROUND: Malunion of the metacarpal mid-shaft fractures are common sequelae and can result in angular deformities of the shaft, bone shortening, or as in most cases, a combination of both in several variations. The degree of the loss of function or efficiency depends on the extent of the malunion. This would also have a bearing as to whether surgical intervention is necessary. METHODS: Experiments were performed on the second metacarpal in eight fresh normal cadaver hands. The extrinsic tendon forces to obtain full flexion and extension of the digit were measured at a fixed tendon excursion. The intact metacarpal was used as the experimental control. An oblique osteotomy was done on the mid-shaft of the metacarpal and combinations of bone shortening (0-5 mm) and dorsal angulation at the mid-shaft of the metacarpal (0-60 degrees ) were simulated. The tendon forces were then measured again as a percentage of the control, given the same excursion. RESULTS: The extension force was found to increase with increasing dorsal angulation and decrease with increasing bone shortening. While the flexion force decreased with increasing dorsal angulation and shortening. Dorsal angulation had a greater effect than bone shortening on the extension and flexion forces. CONCLUSION: Simulated malunion metacarpal fractures in human cadavers, has an effect on the efficiency of the measured extensor and flexor forces. In extension, dorsal angulation was found to increase the force required for the same amount of excursion. However, with added bone shortening, the extension force required decreased. This could be a compensatory mechanism in malunion metacarpal fractures. In flexion, a lower force was required with bone shortening and dorsal angulation for the same excursion. PMID- 11415826 TI - Influence of loading duration on the start-up friction in synovial joints: measurements using a robotic system. AB - OBJECTIVE: To determine how much and why static load influences friction in synovial joints. DESIGN: Start-up coefficient of friction in canine stifles was measured after different duration of static load. BACKGROUND: Previous investigators have shown that friction of cartilage on cartilage contact configurations sharply increases with stationary load duration. This phenomenon has not been confirmed in the entire synovial joint. METHODS:: A system to measure joint friction was designed using a robotic arm. Ten canine stifles from six animals were used. Start-up friction of the femoral condyle on the tibial plateau and femoral condyle on glass plate contact configurations was measured. The glass plate was chosen as a rigid surface where ploughing effect cannot occur. RESULTS: The mean value of the start-up frictional coefficient from femoral condyle on tibial plateau was 0.112 (SD 0.005) at 0 s stationary loading, and sharply increased with the stationary loading duration to 0.313 (SD 0.095) at 1800 s. Those from femoral condyles on glass plate were 0.005 (SD 0.003) at 0 s and 0.457 (SD 0.128) at 1800 s. CONCLUSIONS: Friction in synovial joints sharply increases with duration under static load. The ploughing effect on this increase is slight in friction in canine stifles. RELEVANCE: The lubrication mechanism is worth investigating to understand the pathology of joint diseases. Determining friction behaviour is necessary for the investigation of the lubrication mechanism. PMID- 11415827 TI - Lumbar intradiscal pressure measured in the anterior and posterolateral annular regions during asymmetrical loading. AB - OBJECTIVE: To analyze the effect of asymmetrical loading on intradiscal pressure. DESIGN: Human cadaveric lumbar spines were instrumented with multiple pressure sensors and subjected to external mechanical loads. BACKGROUND: Tears and radial fissures in the posterolateral annulus with no evidence of annular protrusion or nuclear extrusion are frequently observed in non-degenerated intervertebral discs. Cadaveric studies have shown that asymmetrical loads lead to posterolateral herniation. Regional overload may be responsible for a progressive structural weakness of the posterolateral annular fibres. METHODS: Three needles each equipped with three independent pressure sensors were inserted in the midplane of the L(3)/L(4) intervertebral disc (n = 16) in the anterior, right posterolateral and left posterolateral regions. Axial rotation was applied in the upright, flexed and extended positions and the pressures recorded. RESULTS: The largest intradiscal pressure increase was observed in the posterolateral inner annular regions, more so in flexion than extension, when combined with axial rotation. Significant centripetal pressure gradients were found only in the posterolateral needles during the upright and flexed positions. CONCLUSIONS: When applying compression and axial rotation, the posterolateral inner annular zones of the intervertebral disc show high stress peaks and centripetal pressure gradients. Asymmetrical loads (rotation) combined with postural changes in the sagittal plane increase these effects, and may be responsible for a chronic mechanical overload of these regions. RELEVANCE: Our findings suggest a predilection for the posterolateral inner annular regions to mechanical failure, especially under asymmetrical loading. PMID- 11415828 TI - Knee flexion and base of support in asymmetrical handling: effects on the worker's dynamic stability and the moments of the L(5)/S(1) and knee joints. AB - OBJECTIVE: To investigate the effects of knee flexion and of the variations of feet lateral spacing on dynamic stability and on the net joint moments distribution between the back and knees. BACKGROUND: The width of the base of support and knee flexion effects on joint moments in asymmetric handling and especially on the worker's stability have rarely been studied. METHODS: Fourteen healthy male subjects performed an asymmetrical lifting task, using four different techniques: two imposed lateral feet spacings (41 and 57 cm) and two knee flexion amplitudes (slightly and deeply flexed knees). A tridimensional dynamic rigid body model was used to estimate the triaxial net reaction moments at L(5)/S(1) and at the knees, using two force platforms. New developments have been undertaken to characterize workers' stability while handling: the horizontal force required to destabilize the worker was calculated as a measure of dynamic stability. RESULTS: The width of the base of support had little effect on L(5)/S(1) and knee moments; however, the subjects were less stable with the narrow base of support. Using the slightly flexed knees technique, trunk maximal resultant moments were slightly smaller (202 vs. 216 Nm), and maximal resultant knee moments were larger (96 vs. 62 Nm). Furthermore, asymmetric moments at the trunk and the asymmetric position of the knees were reduced with this technique, but subjects were less stable. CONCLUSIONS: The use of a slightly flexed knees technique in asymmetrical handling of low-lying loads appears advantageous because it reduces L(5)/S(1) moments while increasing the knees' flexing moments, although this may compromise workers' stability. RELEVANCE: Handling methods used by workers in asymmetrical handling have rarely been studied. Optimizing the safety of a handling method can involve many parameters, such as reducing joint moments and maximizing stability. The evaluation of the worker's stability while handling in conjunction with joint moments is an interesting alternative to study the safety of handling methods. PMID- 11415829 TI - Dynamic loading on the human musculoskeletal system -- effect of fatigue. AB - OBJECTIVE: A study was conducted to investigate the effects of fatigue on the ability of human musculoskeletal system to deal with the onslaught of the heel strike initiated shock waves. DESIGN: Running on a treadmill at the anaerobic threshold level for 30 min was used to acquire the experimental data on the foot strike initiated shock waves. BACKGROUND: Muscles act to lower the bending stress on bone and to attenuate the dynamic load on human musculoskeletal system. Fatigue may diminish their ability to dissipate and attenuate loading on the system. Knowledge of the effects of fatigue on the ability of the human musculoskeletal system to attenuate the shock waves may help in design of the training procedures and exercises. METHODS: Twenty-two young healthy males participated in this study. Each one was running on the treadmill at the speed corresponding to his anaerobic threshold for 30 min. The heel strike induced shock waves were recorded every 5 min on the tibial tuberosity and sacrum. The data obtained were analyzed in both temporal and frequency domains. RESULTS: The results reveal significant increase in the dynamic loading experienced by the human musculoskeletal system with fatigue. This may be attributed to the inability of the fatigued system to provide an efficient way to attenuate shock waves. CONCLUSIONS: The analysis of the recorded signals suggests that fatigue contributes to the reduction of the human musculoskeletal system's capacity to attenuate and dissipate those shock waves. This capacity appears to be a function not only of the fatigue level, but also of the vertical location along the skeleton. RELEVANCE: Fatigue during running may affect the ability of the human musculoskeletal system to attenuate and dissipate the heel strike induced shock waves. The study of the fatigue effect on shock wave attenuation provides information that may benefit the runner. PMID- 11415830 TI - Passive regulation of impact forces in heel-toe running. AB - OBJECTIVE: the purpose of this study was to determine whether passive mechanisms can account for impact force regulation with changing shoe hardness. DESIGN: A three-dimensional musculoskeletal model of the lower extremity was developed to simulate impact in running with two different shoe hardnesses. BACKGROUND: Considerable research has focused on developing shoe cushioning to reduce impact forces. However, only minimal changes in peak external impact force have been observed with changes in shoe hardness. It is hypothesized that passive mechanisms can regulate impact forces with changing shoe hardness, without changing muscle activities. METHODS: Initial kinematic inputs for the simulations were measured from nine male subjects performing heel-toe running. Simulations were performed with initial conditions and muscle stimulation patterns held constant while shoe hardness was varied between a hard and a soft condition. RESULTS: There was no significant difference between the soft and hard shoe peak impact forces. Peak rates of loading were greater for the hard shoe than the soft shoe. Muscle forces changed with shoe conditions. For some muscles (including the tibialis anterior) the forces were greater for the hard shoe, whereas for other muscles (including the peroneus) forces were greater for the soft shoe condition. CONCLUSIONS: Peak impact forces with changing shoe conditions can be regulated by passive mechanical changes without changing muscle activities or kinematics before touchdown. RELEVANCE: Potential injury causing loads on internal structures (e.g. muscles, tendons, etc.) during the impact phase of running can depend upon shoe hardness, but are not reflected in changes in external ground reaction force. PMID- 11415831 TI - Individual muscle contributions to the in vivo achilles tendon force. AB - OBJECTIVE: To ascertain the possibility of non-uniform stress within the achilles tendon due to individual force contributions of the triceps surae. DESIGN: Calculation of non-uniform stress through discrepancies in moments about the ankle joint. BACKGROUND: Non-uniform stress over the cross-sectional area have been implied in the etiology of achilles tendon injury and may influence functional aspects. However, this has not been empirically demonstrated. METHODS: In vivo achilles tendon forces were measured with an optic fibre technique during isometric plantarflexions at systematically varied knee angles and contraction intensities. A comparison to the plantar force measured underneath the metatarsal heads permitted the calculation of the achilles tendon contribution to the resultant plantarflexion moment. The achilles tendon force was further differentiated into gastrocnemius and soleus contributions. Individual muscle activation patterns were described. RESULTS: The average achilles tendon contribution to the resultant moment was 67.4%. Variations were found at different knee angles and contraction intensities. A force discrepancy of 967 N occurred between gastrocnemius and soleus over a gastrocnemius length change of 2.67 cm. This corresponded to a stress discrepancy of 21 N/mm(2) over the tendon cross-sectional area. Separate muscles showed individual activation patterns. CONCLUSIONS: Non-uniform stress in the achilles tendon can occur through modifications of individual muscle contributions. RELEVANCE: Non-uniform stress in the achilles tendon has been implied in its injury etiology. This study demonstrated such loading resulting from discrepancies in individual muscle forces. PMID- 11415832 TI - A method for the measurement of three-dimensional geometry of casts used for the manufacture of ankle foot orthoses. AB - OBJECTIVE: To develop a method to measure the geometry of casts for the manufacture of ankle foot orthosis. DESIGN: A measurement study in which the feasibility of the technique, and its reliability were studied, using statistics based upon the analysis of variance. BACKGROUND: Studies have shown that the effectiveness of ankle foot orthoses is influenced by small changes in geometry and, particularly, by the amount of dorsiflexion. This study is concerned with a technique to identify the variations between nominally identical casts. METHODS: A measuring technique was used to determine the coordinates of grids drawn on the surface of the casts. Repeatability studies involved the measurement of six casts of the same leg produced by one orthotist. The influence of rectification, in which the technician makes modification prior to moulding, was also studied. RESULTS: The measurement technique was reliable, and there were no significant differences in grid dimensions among the casts. There were significant differences in dorsiflexion (up to 11 degrees) which were not influenced by rectification. CONCLUSIONS: This technique has been shown to be suitable to study the geometry of casts for the manufacture of ankle foot orthoses, and has demonstrated that relatively large variations may occur between nominally identical casts. There is a need to apply this procedure to study the influence of orthotic technique on geometry and function. RELEVANCE: AFO's are prescribed for patients with neurological disabilities. Since it is known that a correctly manufactured orthosis can achieve significant improvements in gait, it is necessary to quantify manufacturing accuracy. PMID- 11415833 TI - Biomechanical evaluation of a central rod system in the treatment of scoliosis. AB - OBJECTIVE: To evaluate the efficiency of a central rod system in the treatment of scoliosis. DESIGN: Biomechanical testing and finite element analysis were performed to compare the stiffness and stress distribution of the central rod system with the Cotrel-Dubousset instrumentation. BACKGROUND: During the fixation and derotation processes in the treatment of scoliosis, the Cotrel-Dubousset instrumentation concentrates unilateral stress on the vertebral elements. It may be possible to cause severe injury to the bony tissues especially in the ostaeoporotic patients. A central rod system was developed to reduce the damage to the bony tissue of the vertebrae in the treatment of scoliosis. METHODS: Mechanical non-destructive cyclical tests comparing the compression, flexion, extension and torsional stiffness were carried out on the models of the central rod system and Cotrel-Dubousset instrumentation using a 858 Bionix test system. The stress distribution on the central rod, the Cotrel-Dubousset rod and a vertebral model consisting in four vertebrae each implanted with two screws was evaluated using a general purpose finite element software ANSYS R 5.0a. The SOLID73 element was employed for three-dimensional modelling. RESULTS: Although the central rod system was found to have lower values in the compression, flexion and extension stiffness, its torsional stiffness is comparable to the Cotrel Dubousset instrumentation. Moreover, the stability of the new design is satisfactory. Finite element analysis indicates that the maximum equivalent stress on the vertebral model with the central rod system is relatively constant. CONCLUSIONS: The central rod system consists of a central rod which applies fixation and derotation through a connector between the screws on both sides of the vertebral column. This new design can not only prevent the effect of unilateral stress concentration, but is also simpler and easier to implant. However, it is necessary to strengthen the material of the rod or the connection between the rod and the screws. RELEVANCE: This study employed the methods of biomechanical testing and finite element analysis to evaluate a newly designed central rod system in the treatment of scoliosis. The same methods can be applied to the evaluation of orthopaedic instrumentations in the pre-clinical stage. The efficiency of the central rod system is comparable to the Cotrel-Dubousset instrumentation, and its implantation is simpler and easier. PMID- 11415834 TI - Data movement in gait analysis for clinical applications. PMID- 11415835 TI - A comparison of peak vs cumulative physical work exposure risk factors for the reporting of low back pain in the automotive industry. AB - OBJECTIVE: To determine the relative importance of modelled peak spine loads, hand loads, trunk kinematics and cumulative spine loads as predictors of reported low back pain (LBP). BACKGROUND: The authors have recently shown that both biomechemical and psychosocial variables are important in the reporting of LBP. In previous studies, peak spinal load risk factors have been identified and while there is in vitro evidence for adverse effects of excessive cumulative load on tissue, there is little epidemiological evidence. METHODS: Physical exposures to peak and cumulative lumbar spine moment, compression and shear forces, trunk kinematics, and forces on hands were analyzed on 130 randomly selected controls and 104 cases. Univariable and multivariable odds ratios of the risk of reporting were calculated from a backwards logistic regression analysis. Interrelationships among variables were examined by factor analysis. RESULTS: Cases showed significantly higher loading on all biomechanical variables. Four independent risk factors were identified: integrated lumbar moment (over a shift), 'usual' hand force, peak shear force at the level of L(4)/L(5) and peak trunk velocity. Substituting lumbar compression or moment for shear did not appreciably alter odds ratios because of high correlations among these variables. CONCLUSIONS: Cumulative biomechanical variables are important risk factors in the reporting of LBP. Spinal tissue loading estimates from a biomechanical model provide information not included in the trunk kinematics and hand force inputs to the model alone. Workers in the top 25% of loading exposure on all risk factors are at about six times the risk of reporting LBP when compared with those in the bottom 25%. RELEVANCE: Primary prevention, treatment, and return to work efforts for individuals reporting LBP all require understanding of risk factors. The results suggest that cumulative loading of the low back is important etiologically and highlight the need for better information on the response of spinal tissues to cumulative loading. PMID- 11415836 TI - Three-dimensional measurement of lumbar spine kinematics for fast bowlers in cricket. AB - OBJECTIVE: To determine whether the three-dimensional (3-D) lumbar spine kinematics for the mixed fast bowling technique differed to those of the side-on and front-on fast bowling techniques. BACKGROUND: It has been previously shown that bowlers who utilise a mixed bowling technique are more likely to show lumbar spine pathology than those who bowl with either the side-on or front-on techniques. METHODS: An electromagnetic device (3-Space(R)Fastrak(TM)) operating at 120 Hz captured range of motion and 3-D lumbar spine kinematics during the delivery stride of 20 young high performance subjects. The trajectory of shoulder and pelvic girdle markers were simultaneously captured and these data were used to classify bowlers into either a side-on, front-on or mixed technique group. RESULTS: No significant differences (P<0.004) existed between the side-on/front on and mixed groups for 12 selected variables derived from the lumbar spine kinematic data. However, an examination of effect sizes revealed evidence that the mixed group showed: a greater amount of left lateral bend and an extended lumbar spine at front foot impact; a body position further from a neutral orientation at lease; and a greater range of motion and angular velocity of the trunk in the lateral bending and flexion/extension axes. CONCLUSIONS: Selected lumbar range of motion and velocity measures tended to be higher for mixed bowlers than side-on/front-on bowlers. RELEVANCE: Overuse injuries to fast bowlers in bricket are common. To better understand the mechanics of injury it is necessary to understand the 3-D rotations of the lumbar spine during this activity. PMID- 11415837 TI - The effect of imposed and self-selected computer monitor height on posture and gaze angle. AB - OBJECTIVE:: The objectives were to determine the postural consequences of varying computer monitor height and to describe self-selected monitor heights and postures. DESIGN:: The design involved experimental manipulation of computer monitor height, description of self-selected heights, and measurement of posture and gaze angles. BACKGROUND: Disagreement exists with regard to the appropriate height of computer monitors. It is known that users alter both head orientation and gaze angle in response to changes in monitor height; however the relative contribution of atlanto-occipital and cervical flexion to the change in head rotation is unknown. No information is available with regard to self-selected monitor heights. METHODS: Twelve students performed a tracking task with the monitor placed at three different heights. The subjects then completed eight trials in which monitor height was first self-selected. Sagittal postural and gaze angle data were determined by digitizing markers defining a two-dimensional three-link model of the trunk, cervical spine and head. RESULTS: The 27 degrees change in monitor height imposed was, on average, accommodated by 18 degrees of head inclination and a 9 degrees change in gaze angle relative to the head. The change in head inclination was achieved by a 6 degrees change in trunk inclination, a 4 degrees change in cervical flexion, and a 7 degrees change in atlanto-occipital flexion. The self-selected height varied depending on the initial monitor height and inclination. CONCLUSIONS: Self-selected monitor heights were lower than current 'eye-level' recommendations. Lower monitor heights are likely to reduce both visual and musculoskeletal discomfort. RELEVANCE: Musculoskeletal and visual discomfort may be reduced by placing computer monitors lower than currently recommended. PMID- 11415838 TI - Velocity effects on the scapulo-humeral rhythm. AB - OBJECTIVE: The objective of this study is to verify the assumption that the three dimensional (3-D) shoulder motions can be described by means of an interpolation of statically recorded postures and thus, support the application of non-invasive but static techniques for motion analysis of the shoulder. BACKGROUND: During shoulder motions the scapula moves underneath the skin. Recording of motions is only possible by means of invasive methods. An alternative for the recording is palpation of skeletal landmarks on the scapula and subsequent digitization. The method is non-invasive and relatively easy, but static. Motions are modelled by means of interpolation of the subsequent position recordings. Validity of this method, however, has never been demonstrated. METHODS: Seven subjects performed an alternating abduction-adduction motion of the arm in a plane 30 degrees forward rotated with respect to the frontal plane, at three sub-maximal frequencies: 0.04, 0.25 and 0.50 Hz. The humeral and scapular motions were recorded by means of a two-dimensional (2-D) X-ray video system. The motions of the humerus, the scapular spine and the glenoid ridge were defined by angles, and the sinusoidal motion curves were characterized by means of the offset, the amplitude and the phase of the motions. RESULTS: By means of Repeated Measurements Multi-Variate Analysis of Variance, a significant effect of arm motion on the phase and the amplitude of the scapular motion was found. However, the magnitude of the effects are negligibly small for the present applications at sub-maximal arm motion velocities. CONCLUSIONS: For normal arm motions in the vertical plane, the kinematics of the shoulder skeleton can be derived by the interpolation of statically recorded positions of the bones. RELEVANCE: The 3-D motions of the shoulder are the result of the kinematic constraints of the skeletal system and the coordinated muscle forces, and are only one of the few characteristics that can be quantified. The motions contain relevant information which is essential in the analysis of clinical disorders, e.g. sub-acromial disorders and glenohumeral subluxation, the evaluation of clinical interventions and physiotherapy, and in the analysis of ergonomic and biomechanical problems. PMID- 11415839 TI - Is the stability of a tibial fracture influenced by the type of unilateral external fixator? AB - OBJECTIVE: To examine whether the type of unilateral external fixator significantly influences the stability of a tibial fracture. DESIGN: Inter fragmentary displacements were measured during walking while the fractures were stabilized, first with one type of fixator then with another. BACKGROUND: It is commonly claimed that one type of fixator exerts a different influence on mechanical stability at a fracture in comparison with another. METHODS: This study compares inter-fragmentary displacement, fixator displacement and weight bearing during walking, in four patients stabilized with an Orthofix DAF, which was replaced by a Howmedica International Monotube. RESULTS: The null hypothesis of no difference in fixator performance was unproved (P < 0.05) through insufficient data. Interpatient variability in inter-fragmentary displacement implies that anthropometry, gait and fracture type may influence fracture stability more than the type of fixator. CONCLUSIONS: Since weight-bearing and displacements were not substantially different, no basis was found for the claim that one fixator provides a mechanical environment substantially different to another. Sample size was not large enough to prove that a small but statistically significant difference exists. RELEVANCE: The influence on fracture stability of one type of unilateral fixator in comparison with another appears to be less than the influence of anthropometry, gait and the type of fracture. PMID- 11415840 TI - In vitro testing of a novel limb salvage prosthesis for the distal femur. AB - OBJECTIVE: The aim of this paper was to define strain pattern in the host bone following distal femoral resection and implantation of a massive prosthesis. Two methods of coupling the prosthesis to the bone were compared: the Compliant Pre Stress device, and a standard cemented tumour prosthesis. DESIGN: The composite femur model was selected to minimize variables. Four femurs were tested before and after implantation. Both coaxial and cantilever loading were applied. BACKGROUND: Cemented distal femoral replacement following resection of malignant tumours has a high failure rate at 5 years and is associated with extensive bone resorption thought to be secondary to stress shielding. METHODS: Strain was measured in the medial and lateral sides at four levels with physiologic loads applied, in the intact, Compliant Pre-Stress, and cemented femurs. Repeated measurements were taken. Strains in the implanted femur were calculated as percentage of the intact, and statistically analyzed. RESULTS: The most reproducible results were noted in cantilever bending (variability <5%). The Compliant Pre-Stress device demonstrated a more physiologic strain pattern than the cemented stem. The most significant difference between the two implants was in the area adjacent to the interface. CONCLUSIONS: The Compliant Pre-Stress device shows less stress shielding than a standard cemented implant. The protocol described and the use of composite femurs demonstrated reproducible results. RELEVANCE: Massive prosthesis are commonly used following tumour resection or removal of failed primary joint replacement prostheses. The failure rate for aseptic loosening for cemented implants is 25% at 5 years with significant bone resorption about the implant. Compliant Pre-Stress is an innovative technology that allows coupling of metallic implants to bone with little stress shielding. This paper aims to define the strain patterns about the implant and compare them to a standard cemented device. The reduced stress shielding of the Compliant Pre Stress fixation system should guarantee reduced bone loss around the implant and help to obtain improved clinical results. PMID- 11415841 TI - The effects of axial and multi-plane loading of the extensor mechanism on the patellofemoral joint. AB - OBJECTIVE: To compare the effects of axial loading, and anatomically based multi plane loading of the extensor mechanism on the patellofemoral joint. DESIGN: Repeated measures design using an in-vitro cadaver model. BACKGROUND: Since the extensor mechanism is the primary contributor to the patellofemoral joint reaction force and can affect patellar kinematics, it is essential that the forces produced by this musculature be accurately represented in a simulation model. METHODS: Patellar kinematics (magnetic tracking device), contact pressures and areas (pressure sensitive film) were measured from 6 cadaver knees under two different loading conditions: 1) axial (rectus femoris loaded in the frontal plane), and 2) multiplane (individual components of the quadriceps loaded along their respective fiber directions in both the frontal and sagittal planes). Specimens were mounted in a custom knee jig, with muscle forces being simulated using a pulley system and weight. Data were collected at 0 degrees, 15 degrees, 30 degrees, 45 degrees, 60 degrees, 75 degrees and 90 degrees of knee flexion. RESULTS: Compared to the axial loading condition, multi-plane loading of the vasti resulted in significantly greater contact pressure at 0 degrees and significantly less contact pressure at 90 degrees of knee flexion. Furthermore, the multi-plane loading condition resulted in greater lateral patellar rotation from 0-75 degrees of knee flexion, and greater lateral glide at 30 degrees of knee flexion. Greater patellar flexion was observed with the axial loading condition. CONCLUSIONS: These findings indicate that axial loading of the extensor mechanism underestimates contact pressure at 0 degrees and overestimates contact pressure at 90 degrees of knee flexion when compared to multi-plane loading. Additionally, loading of the individual vasti appears to have an effect on patellar kinematics. RELEVANCE: The results of this study indicate that anatomically based, multi-plane loading of the vasti will yield subtle yet significant differences in patellofemoral joint mechanics when compared to the more traditional axial loading approach. These differences may have implications for the study of both normal and pathological patellofemoral joint mechanics, as well as evaluation of surgical techniques and prosthetic implants. PMID- 11415842 TI - Biomechanical response of the passive human knee joint under anterior-posterior forces. AB - OBJECTIVE: To investigate the detailed biomechanics of the passive tibiofemoral knee joints in full extension under anterior/posterior drawer forces of up to 400 N. DESIGN: A nonlinear three-dimensional finite element model of the entire human tibiofemoral joint consisting of bony structures, their articular cartilage layers, menisci, and four principal ligaments was utilized. BACKGROUND: The mechanics of the knee joint, specially under drawer forces, have extensively been investigated. Despite all these works, the detailed joint biomechanics, specially the role of boundary conditions, load transmission through menisci/articular cartilage layers, and coupling between menisci and cruciate ligaments are not yet quantified. METHODS: Nonlinear elastostatic analyses were carried out considering the tibiofemoral joint at full extension under anterior and posterior loads of up to 400 N applied either to the tibial or the femoral shaft. Cases with various boundary conditions, cruciate ligament deficiency (anterior or posterior), and total unilateral meniscectomy (medial or lateral) were analysed. RESULTS: In addition to the total primary anterior-posterior motion of about 9 mm at +/-400 N, significant coupled external tibial rotations of about 9 degrees and 10 degrees were computed under 400 N femoral posterior and anterior forces, respectively. The response was influenced by the manner of loading and boundary conditions. The anterior cruciate ligament and posterior cruciate ligament were the primary restraints to femoral posterior and anterior drawer forces, respectively. Section of either of these ligaments drastically increased the joint anterior-posterior motion. In the absence of cruciates, the collaterals became the primary restraints in both anterior-posterior forces. In this case, the tibial plateaus, specially the medial one in the anterior cruciate ligament deficient joint, experienced much larger compressive forces. In addition to causing an increase in joint primary anterior-posterior laxity and anterior cruciate ligament forces, medial meniscectomy substantially increased coupled tibial external rotation, forces on the lateral plateau, and stresses in the articular cartilage of the lateral plateau. RELEVANCE: Our results suggest an increased role for the medial meniscus in the anterior cruciate ligament deficient joint. Lateral meniscectomy had much smaller effects on results than the medial one. The success of any anterior cruciate ligament replacement or meniscal transplantation in the restoration of the joint stability and the protection of the articular cartilage against excessive stresses depends on the coupling between the anterior cruciate ligament and medial meniscus. Absence of any of these components would drastically influence the joint response. PMID- 11415843 TI - Morphological changes of the triceps surae muscle-tendon unit during passive extension: an in vivo rabbit model. AB - OBJECTIVE: To elucidate the morphological and biomechanical manifestation of the triceps surae muscle-tendon unit during passive extension. DESIGN: The instantaneous changes within the load-deformation curve of muscle-tendon unit were analyzed by an in vivo rabbit model. BACKGROUND: Although muscle strains occur more frequently than complete failures, the failure mechanism of these sub failure injuries is rarely investigated. Monitoring of the instantaneous changes in the load-deformation curve allows correlation with the morphological changes that occur during passive extension. METHODS: After anesthesia, the triceps surae muscle of rabbit was dissected and then stretched to failure by a MTS Bionix 858 machine. The morphological changes in failure patterns were recorded by photographs. RESULTS: The morphological and biomechanical manifestations of the triceps surae muscle-tendon unit was divided into five different portions: first, the viscoelastic portion with minimal morphologic change; second, the portion of micro-failure with local ecchymosis; third and fourth, the portions of macrofailure with sequential rupture of the muscle fibers; and fifth, the portion of rupture and separation of muscle parenchyma. CONCLUSIONS: A threshold for stretch-induced injury does exist. The threshold of the initiation of micro failure in this model was 16.5% of the strain, which corresponded to 16.6% of the maximal sustainable force. RELEVANCE: Elucidation of the existence of micro failure in skeletal muscle gives a good reference point to determine the functional capabilities and limitation of muscular tissue. With this, the deleterious effect of muscle stretching can be avoided by limiting muscle stretching to below this limit. PMID- 11415844 TI - ISB Clinical Biomechanics Award 1997. PMID- 11415845 TI - Effect of ovariectomy and calcium deficiency on the ultrasound velocity, mineral density and strength in the rat femur. PMID- 11415846 TI - Age, body mass index, race and other determinants of steroid hormone variability: the HERITAGE Family Study. AB - OBJECTIVE AND METHODS: To investigate from the HERITAGE Family Study database, 13 steroid hormones (androstane-3alpha, 17beta-diol glucuronide, androsterone glucuronide, cortisol, dehydroepiandrosterone (DHEA), DHEA ester (DHEAE), DHEA sulfate (DHEAS), dihydrotestosterone (DHT), estradiol, 17-hydroxyprogesterone, progesterone, pregnenolone ester, sex hormone binding globulin (SHBG) and testosterone in each sex for their relationships with age, body mass index (BMI), race and key lifestyle variables. Sample sizes varied from 676 to 750 per hormone. Incremental regression methods were used to examine the contributions of the variables to steroid hormone variability. RESULTS: Age was a major predictor for most steroid hormones. The greatest contribution of age was a negative relationship with DHEAS (R(2)=0.39). BMI was also associated with the variability of several steroid hormones, being the most important predictor of SHBG (R(2)=0.20) and of testosterone (R(2)=0.12) concentrations. When age and BMI were included, race still contributed significantly to the variations in cortisol (R(2)=0.02 for men and 0.04 for women), DHT (R(2)=0.02 for men and 0.03 for women), and progesterone (R(2)=0.03 for women). Nevertheless, race appeared to be less important than age and BMI. In addition, lifestyle indicators (food and nutrient intakes, smoking and physical activity) influenced steroid hormone variability. Their contributions, however, were minor in most cases once age, BMI and race had been taken into account. CONCLUSIONS: We conclude that age was the most important factor, followed by BMI, race and lifestyle factors in explaining steroid hormone variability. PMID- 11415847 TI - Impact of protein energy malnutrition on thyroid size in an iodine deficient population of Gujarat (India): Is it an aetiological factor for goiter? AB - OBJECTIVE: To assess the severity of protein energy malnutrition (PEM) in iodine deficient subjects and to assess the impact of PEM on thyroid size. METHODS: 1002 subjects (530 school-aged children and 472 adults) were assessed for PEM by direct anthropometric measurements of height, weight, triceps skinfold (TSF) thickness, mid upper arm circumference (MUAC) and thigh circumference (TC), and derived indices of body surface area (BSA), body mass index (BMI), and Z-scores for weight-for-age (WAZ), height-for-age (HAZ), and weight-for-height (WHZ). Severity of PEM was based on the World Health Organization (WHO) criteria and the threshold on the Waterlow classification. Thyroid size was measured by ultrasonography to determine the thyroid volume (TV). Linear regression analysis was performed between TV and anthropometric parameters. RESULTS: Children had severe PEM as evident from the WHO percentage prevalence of stunting (HAZ< 2SD)=64% (where <-2SD is the Z-score deficit), wasting (WHZ<-2SD)=43%, underweight (WAZ<-2SD)=82% and BMI<16 kg/m=90%. Waterlow classification showed that children were either stunted or wasted, or stunted and wasted, or stunted and obese. Nearly 100% (529/530) of the children had goiter as evidenced from enlarged TV-for-BSA when compared with the WHO reference. There was a weak but statistically significant (P<0.05) positive correlation between TV and BSA, weight, height, MUAC, TC and HAZ but a negative correlation between TV and WHZ, BMI and TSF (r=-0.1-0.2). Adults had PEM as evident from BMI<18.5 kg/m in 54% subjects. Median MUAC=22.7 cm reveals prolonged severe PEM. Eighty-two percent had enlarged TV (>20 ml). There was a significant (P=0.01) negative correlation between TV and MUAC. CONCLUSIONS: (i) The severity of acute (wasting) and chronic (stunting) PEM is very high in Gujarati children. They are stunted or wasted, or stunted and wasted, or stunted and obese. Gujarati adults are thin with low protein and fat reserves. (ii) Anthropometric parameters showed a significant (P<0.001) correlation (r=0.1-0.2) with thyroid size. (iii) Higher prevalence of goiter may be due to macro-nutrient malnutrition (PEM) in the face of micro nutrient malnutrition (iodine deficiency disorders, IDD). PMID- 11415848 TI - Novel mutations of the thyroid peroxidase gene in patients with permanent congenital hypothyroidism. AB - OBJECTIVE: It is suggested that iodide organification defects account for 10% of all cases with congenital hypothyroidism (CH). One candidate gene for these defects is the thyroid peroxidase (TPO) gene. DESIGN: Exons 2, 8-10 and 14 of the TPO gene were examined in 30 patients with permanent CH without a family history of CH. This group was characterized by the presence of an orthotopic thyroid gland and elevated TSH levels. METHODS: The mutational screening was performed by single-strand conformational polymorphism followed by sequence analysis of fragments with abnormal migration patterns and by restriction enzyme analysis. RESULTS: In four patients we were able to identify mutations on both alleles which have not been described so far. One patient was a carrier of a new homozygous point mutation in exon 9 resulting in an exchange from Leu to Pro at codon 458. Another patient was found to be compound heterozygous for two mutations, a 20 bp duplication in exon 2 and a new mutation in exon 9 (Arg491His). Two brothers of consanguineous parents showed a homozygous T deletion in exon 14 at position 2512. CONCLUSIONS: Our findings confirm the genetic heterogeneity of TPO defects and support the suggested prevalence of organification defects. PMID- 11415849 TI - Neuroactive steroid-serotonergic interaction: responses to an intravenous L tryptophan challenge in women with premenstrual syndrome. AB - OBJECTIVE: To evaluate the circulating concentrations of the neuroactive steroids in response to an i.v. L-tryptophan (L-TP) challenge across the menstrual cycle in women with premenstrual syndrome (PMS) and in controls. METHOD: An i.v. L-TP challenge was administered eight times during 1 month to five women with prospectively documented PMS and five age- and body mass-matched controls. Progesterone, allopregnanolone pregnenolone and 3alpha-5alpha tetrahydrocorticosterone were assessed 15 and 0 min before, and at 30, 60 and 90 min after the challenge, across the menstrual cycle. RESULTS: In response to L-TP challenge, only allopregnanolone concentrations were significantly increased across the cycle and this increase was of a greater magnitude in women with PMS. Pregnenolone and 3alpha-5alpha-tetrahydrocorticosterone concentrations were not affected in women with PMS or controls after L-TP challenge. CONCLUSIONS: The data provide evidence for possible interaction between the serotonergic system and the neuroactive steroid, allopregnanolone. Women with PMS demonstrated a more significant increase in allopregnanolone concentrations in response to L-TP challenge, which could be due to an initial low basal serotonergic tone in the luteal phase in the PMS group. PMID- 11415850 TI - A luteinizing hormone-, alpha-subunit- and prolactin-secreting pituitary adenoma responsive to somatostatin analogs: in vivo and in vitro studies. AB - OBJECTIVE: Evaluation of the efficiency of somatostatin analogues in the treatment of a mixed luteinizing hormone (LH)-, alpha-subunit-, prolactin (PRL) secreting pituitary adenoma. DESIGN: A 30-year-old woman, with amenorrhaea galactorrhaea, presented with a pituitary macroadenoma. The endocrine evaluation showed high plasma levels of PRL, LH, and alpha-subunit inhibited by 65%, 65% and 33% respectively under octreotide test (200 microg, s.c.). Long-term treatment with slow release (SR) lanreotide (30 mg/10 days, i.m.) restored menstrual cycles and normalized PRL values. Due to persisting supranormal levels of LH and alpha subunit, and to the absence of tumoral shrinkage, the adenoma was resected by the transsphenoidal route. METHODS: In vitro characterization of the somatostatin receptor subtypes (SSTR) expression and functionality. Real-time polymerase chain reaction was performed to quantify the expression of SSTR mRNAs and functionality of the SSTRs was assessed in cell culture studies with various concentrations of native somatostatin (SRIF-14) and of analogues preferential for SSTR2 or SSTR5. RESULTS: This adenoma presented with high levels of SSTR2, SSTR3 and SSTR5 mRNAs, as compared with a series of gonadotroph adenomas. In cell culture studies, PRL, LH and alpha-subunit were inhibited by 60%, 47% and 33% respectively by SRIF-14 at a concentration of 10 nmol/l. The SSTR2 (BIM-23197, lanreotide) and SSTR5 (BIM 23268) preferential analogues both produced a partial 21-38% inhibition of PRL, LH, and alpha-subunit release. DISCUSSION: In this plurihormonal-secreting adenoma, the high efficacy of somatostatin analogues to inhibit PRL, LH and alpha subunit secretion in vivo may be explained by the unusually high level of expression and by the functionality of both SSTR2 and SSTR5 receptor subtypes. PMID- 11415851 TI - Leptin levels show diurnal variation throughout puberty in healthy children, and follow a gender-specific pattern. AB - OBJECTIVE: To investigate the levels and diurnal rhythm of serum leptin in healthy children, and to investigate the association between leptin levels and sex steroids. METHODS: Four girls and four boys, all healthy volunteers, were followed longitudinally throughout puberty. Their chronological ages ranged from 8.7 to 19.5 years, and body composition, expressed as weight-for-height standard deviation scores (SDS), ranged between -1.7 and +2.4. Serum leptin, oestradiol and testosterone concentrations were measured by radioimmunoassay at 1000, 1400, 1800, 2200, 0200 and 0600 h. RESULTS: In all girls and boys, both prepubertally and during pubertal development, serum leptin levels increased during the night, with no difference in relative peak amplitude. In boys, the leptin concentrations increased until the initiation of puberty and then declined, whereas in girls, the concentrations increased throughout puberty. The inter-individual variation in mean leptin levels among girls decreased to 11% at the time of menarche. A positive correlation was found for both oestradiol and testosterone versus leptin in girls throughout puberty (r=0.64 and r=0.71 respectively, P<0.001). A negative correlation was found between leptin and testosterone in boys in mid- and late puberty (r=-0.66, P<0.01). No correlation was found between oestradiol and leptin in boys or between testosterone and leptin in pre- and early pubertal boys. CONCLUSION: Serum leptin concentrations show diurnal variation throughout pubertal development in both girls and boys. The changes in leptin levels during puberty follow a gender-specific pattern, probably due to an influence of sex steroids on leptin production. PMID- 11415852 TI - Body mass, plasma leptin, glucose, insulin and C-peptide in offspring of diabetic and non-diabetic mothers. AB - OBJECTIVE: The aim was to investigate the relationship between body mass index (BMI), plasma leptin, glucose, insulin and C-peptide levels in the offspring of diabetic mothers (DM) and non-diabetic healthy mothers (HM). DESIGN: Seventy-two offspring (37 girls and 35 boys, age 4-20 years) of DM were investigated in a prospective study. Those 14-16 years old (Tanner stage II-IV) were compared with age-matched offspring of HM (33 girls and 33 boys). RESULTS: BMI strongly correlated with plasma leptin concentration in the offspring of both DM and HM children. There were higher BMI and plasma leptin and glucose levels in DM than in HM children. There was no difference in plasma insulin or C-peptide levels between HM and age-matched DM children. There was a highly significant positive correlation between plasma leptin and C-peptide in boys of DM. CONCLUSIONS: The higher plasma leptin found in the offspring of DM reflects their higher BMI. A moderately high but still normal glycemia might be a preclinical sign of insulin resistance or other disturbance of glucoregulation. PMID- 11415853 TI - Desethylamiodarone antagonizes the effect of thyroid hormone at the molecular level. AB - OBJECTIVE: To evaluate the molecular mechanisms of the inhibitory effects of amiodarone and its active metabolite, desethylamiodarone (DEA) on thyroid hormone action. MATERIALS AND METHODS: The reporter construct ME-TRE-TK-CAT or TSHbeta TRE-TK-CAT, containing the nucleotide sequence of the thyroid hormone response element (TRE) of either malic enzyme (ME) or TSHbeta genes, thymidine kinase (TK) and chloramphenicol acetyltransferase (CAT) was transiently transfected with RSV TRbeta into NIH3T3 cells. Gel mobility shift assay (EMSA) was performed using labelled synthetic oligonucleotides containing the ME-TRE and in vitro translated thyroid hormone receptor (TR)beta. RESULTS: Addition of 1 micromol/l T4 or T3 to the culture medium increased the basal level of ME-TRE-TK-CAT by 4.5- and 12.5 fold respectively. Amiodarone or DEA (1 micromol/l) increased CAT activity by 1.4 and 3.4-fold respectively. Combination of DEA with T4 or T3 increased CAT activity by 9.4- and 18.9-fold respectively. These data suggested that DEA, but not amiodarone, had a synergistic effect with thyroid hormone on ME-TRE, rather than the postulated inhibitory action; we supposed that this was due to overexpression of the transfected TR into the cells. When the amount of RSV TRbeta was reduced until it was present in a limited amount, allowing competition between thyroid hormone and the drug, addition of 1 micromol/l DEA decreased the T3-dependent expression of the reporter gene by 50%. The inhibitory effect of DEA was partially due to a reduced binding of TR to ME-TRE, as assessed by EMSA. DEA activated the TR-dependent down-regulation by the negative TSH-TRE, although at low level (35% of the down-regulation produced by T3), whereas amiodarone was ineffective. Addition of 1 micromol/l DEA to T3-containing medium reduced the T3 TR-mediated down-regulation of TSH-TRE to 55%. CONCLUSIONS: Our results demonstrate that DEA, but not amiodarone, exerts a direct, although weak, effect on genes that are regulated by thyroid hormone. High concentrations of DEA antagonize the action of T3 at the molecular level, interacting with TR and reducing its binding to TREs. This effect may contribute to the hypothyroid-like effect observed in peripheral tissues of patients receiving amiodarone treatment. PMID- 11415854 TI - Renal effects of recombinant prolactin in anaesthetized rats. AB - OBJECTIVE: To re-examine the controversial possibility that prolactin exerts renal effects, using recombinant mouse prolactin (rmP), in the presence and absence of circulating vasopressin. DESIGN: In experiment 1, the renal effects of rmP were examined in anaesthetized Brattleboro rats with hereditary hypothalamic diabetes insipidus (BDI) lacking circulating vasopressin and normal animals of the parent Long Evans (LE) strain. In experiment 2, salt and water excretion were studied in fluid-loaded normal Sprague-Dawley (SD) rats, some of which received rmP. METHODS: In experiment 1, BDI and LE rats maintained in fluid balance were infused i.v. with each of three concentrations of rmP (10, 20 and 40 microg/ml per h) or maintained on 150 mmol/l NaCl vehicle (controls). In experiment 2, the SD rats were infused with 75 mmol/l NaCl in order to induce a state of diuresis comparable to that of BDI rats, some of them then receiving the rmP i.v. RESULTS: A profound rmP-induced dose-dependent decrease in urine excretion (P<0.005) and a lesser decrease in sodium excretion in the BDI rats was in marked contrast with the small but significant increase in urine excretion in the LE rats compared with controls (P<0.025). The rmP-infused fluid-loaded SD rats also demonstrated a significant (P<0.05) dose-related antidiuresis compared with the control animals, in addition to a decrease in sodium excretion. CONCLUSIONS: These results show that prolactin has a profound antidiuretic effect in the absence of circulating vasopressin. In contrast, when vasopressin is present in the circulation rmP has a small, but opposite, diuretic effect. Thus the use of a recombinant prolactin has provided evidence for renal effects of this hormone which are modified in the presence of the circulating neurohypophysial hormone vasopressin. PMID- 11415855 TI - Sensitivity to exogenous GH and reversibility of the reduced IGF-I gene expression in aging rats. AB - BACKGROUND: IGF-I gene expression and IGF-I plasma concentration decline with age. A decreased sensitivity to GH has been suggested to be a contributory mechanism to this, in addition to attenuated GH secretion. OBJECTIVE: This study focuses on the sensitivity to exogenous GH and the reversibility of the reduced IGF-I gene expression in aging male rats. DESIGN: Three groups of male Wistar rats aged 3 months (young adult), 11 months (middle-aged) and 27 months (old), received recombinant human GH (rhGH) (150 microg/12 h s.c.) for seven consecutive days. RESULTS: This rhGH treatment completely reversed plasma immunoreactive IGF I (IR-IGF-I) and hepatic IGF-I mRNA levels in 11-month-old and 27-month-old animals to the levels of the young group of animals. The sensitivity in the old group (percentage of increment after the same or lower dose of rhGH per body weight) was increased for both parameters; serum IGF-I increment: 15% in 3-month old, 42.6% in 11-month-old and 119.1% in 27-month-old rats; and hepatic IGF-Ib mRNA increase: 45% in 3-month-old, 27.8% in 11-month-old and 64.3% in 27-month old rats. IGF binding protein-3 (IGFBP-3) mRNA level in the liver was significantly decreased in the old group and only a partial reversion occurred in this group after rhGH treatment; the percentage of increment was also higher in the old group of rats. In extrahepatic tissues IGF-I mRNA was not significantly different in the kidney and the testis of the three groups, and the rhGH treatment produced a significant and similar increase of IGF-I mRNA level in the kidney of the three groups of rats and in the testis of the 27-month-old animals. The GHr/GHBP mRNA remained unchanged in the liver and in the kidney or the testis of the three groups, and was not influenced by the rhGH treatment. Exogenous rhGH decreased pituitary GH mRNA accumulation in a more intense manner in the old group versus control of each group: young adult, 25%; middle-aged, 41.2%; and old rats, 55%. The action of rhGH on pituitary immunoreactive GH (IR-GH) content was only significantly evident in the young group. CONCLUSIONS: These results establish that exogenous rhGH recovers the attenuated liver IGF-I gene expression and the diminished plasma IR-IGF-I in old rats to the levels of young adult animals. They also indicate that the hepatic and extrahepatic (kidney and testis) sensitivity to one established dose per weight of exogenous rhGH is not altered in old animals, or could be potentially increased in some tissues. PMID- 11415856 TI - Oestradiol and progesterone change beta3-adrenergic receptor affinity and density in brown adipocytes. AB - OBJECTIVE: To check if the oestradiol- and progesterone-driven reduction in noradrenaline responsiveness of brown adipocytes is due to a reduction in either the density or the affinity of beta3-adrenoceptors (beta3-AR). beta1/beta2-AR were also studied. DESIGN: Four groups of animals were considered. (i) control rats at thermoneutrality, (ii) cold-acclimated rats, to determine beta-AR under continuous sympathetic stimulation, which is known to decrease noradrenaline responsiveness, (iii) oestradiol- and (iv) progesterone-treated cold-acclimated rats to determine hormonal effects on beta-AR populations in thermogenically active brown adipocytes. METHODS: Oestradiol and progesterone were chronically elevated by means of s.c. Silastic implants. Densities and affinities of beta-AR populations were determined by binding studies using [3H]CGP-12177 as radioligand. RESULTS: Two populations of low and high binding affinities (K(d) 1.6 and 27.3 nmol/l) corresponding to beta3- and beta1/beta2-AR respectively were found at thermoneutrality. beta3-AR density was higher than that of beta1/beta2 AR (B(max) 419 and 143 fmol/mg protein respectively). Cold-acclimated rats showed a reduction of beta3-AR binding capacity (B(max) 308 fmol/mg protein). Oestradiol and progesterone reduced the density of beta3-AR to 167 and 185 fmol/mg protein respectively, while increasing their affinity for [3H]CGP-12177 (K(d) 9.5 and 4.0 nmol/l vs 16 nmol/l in cold-acclimated untreated rats). The density of beta1/beta2-AR was also reduced after oestradiol treatment (B(max) 51 fmol/mg protein). CONCLUSIONS: Both oestradiol and progesterone reduce the density of beta3-AR in brown adipose tissue (BAT) while increasing their affinity for [3H]CGP-12177. Oestradiol also reduces the density of beta1/beta2-AR whereas cold acclimation reduces the density of beta3-AR. PMID- 11415857 TI - Adipocyte-specific reduction of phosphodiesterase 3B gene expression and its restoration by JTT-501 in the obese, diabetic KKAy mouse. AB - OBJECTIVE: Phosphodiesterase (PDE) 3B is a key enzyme involved in the anti lipolytic action of insulin in adipocytes. PDE3B activation results in a reduced output of free fatty acids (FFA), whereas elevated serum FFA is known to cause insulin resistance. We have recently reported that reduced PDE3B gene expression is restored by treatment with pioglitazone, in the adipose tissues of obese, insulin-resistant diabetic KKAy mice. To determine whether the altered PDE3B gene expression is specific for adipocytes, the expression of this gene in liver and epididymal fat tissues of KKAy mice was examined. The effect of JTT-501, another peroxisome proliferator-activated receptor (PPAR)gamma ligand, which is different from thiazolidinedione, was also examined. METHODS: PDE3B mRNA and protein were quantified by an RNase protection assay and Western blotting respectively. Membrane-bound PDE activities were also measured. RESULTS: In adipose tissues of KKAy mice, PDE3B mRNA, protein and membrane-bound PDE activity were reduced to 47%, 57% and 51% respectively relative to those in C57BL/6J control mice. JTT-501 increased PDE3B mRNA, protein and membrane-bound PDE activity by 2.2-, 1.6- and 1.7-fold respectively over those of untreated KKAy mice. In the liver, PDE3B gene expression remained unchanged in KKAy mice, and was not affected by JTT-501. JTT 501 reduced the elevated levels of serum insulin, glucose, FFA and triglyceride in KKAy mice. CONCLUSIONS: PDE3B gene expression was specifically reduced in the adipose tissues of KKAy mice. JTT-501 restored this reduced gene expression with an accompanying improvement in elevated serum FFA and insulin resistance. PMID- 11415858 TI - Localized granuloma annulare associated with autoimmune thyroid disease in a patient with a positive family history for autoimmune polyglandular syndrome type II. PMID- 11415859 TI - Increased glucose-dependent insulinotropic polypeptide (GIP) secretion in acromegaly. AB - OBJECTIVE: Acromegaly is often associated with fasting and postprandial hyperinsulinemia, and the mechanisms involved are only partly understood. Hypersecretion of incretins such as glucose-dependent insulinotropic polypeptide (GIP) could play a role in determining hyperinsulinemia in acromegaly, but the available data are inconsistent. The aim of this study was to characterize the fasting and postprandial pattern of plasma GIP and insulin in a group of acromegalic patients. DESIGN AND METHODS: Eleven non-diabetic patients with newly diagnosed acromegaly and 11 sex- and age-matched healthy subjects were studied. Blood samples were taken at regular intervals in fasting conditions and for 3 h after a standard solid-liquid meal for growth hormone (GH), GIP and insulin measurements. RESULTS: Not only insulin, but also fasting and postprandial GIP levels were significantly higher in the patients with acromegaly than the healthy subjects (P<0.01). In the former group fasting GIP levels and the integrated GIP response to the meal correlated significantly with GH basal levels (r=0.83, P<0.01 and r=0.65, P<0.05, respectively). Moreover, multivariate linear regression analysis showed that the presence of acromegalic status was associated with higher fasting and postprandial GIP levels independently of sex, age, fasting and postprandial plasma glucose and insulin levels, and the occurrence of normal or impaired glucose tolerance. CONCLUSION: This study provides evidence that in patients with acromegaly fasting and postprandial GIP levels are abnormally high. GIP hypersecretion in turn might play a role in the pathogenesis of hyperinsulinemia that characterizes acromegaly. PMID- 11415888 TI - Is there a role for diffusion-weighted imaging in patients with brain tumors or is the "bloom off the rose"? PMID- 11415889 TI - Intraarterial signal on fluid-attenuated inversion recovery images: a measure of hemodynamic stress? PMID- 11415890 TI - What role does functional MR imaging play in the diagnosis or prediction of future-onset Alzheimer's disease? PMID- 11415891 TI - CT perfusion flow assessment: "up and coming" or "off and running"? PMID- 11415892 TI - Fluid-attenuated inversion recovery intraarterial signal: an early sign of hyperacute cerebral ischemia. AB - BACKGROUND AND PURPOSE: Early detection of arterial occlusion and perfusion abnormality is necessary for effective therapy of hyperacute cerebral ischemia. We attempted to assess the utility of the fast fluid-attenuated inversion recovery (fast-FLAIR) sequence in detecting occluded arteries as high signal (referred to as intraarterial signal) and to establish the role of fast-FLAIR in detecting ischemic penumbra of hyperacute stroke within 24 hours after ictus. METHODS: We studied 60 patients with hyperacute cerebral ischemia caused by occlusion of intracranial major arteries. We compared intraarterial signal on FLAIR images with time of flight (TOF) on MR angiograms, flow voids on T2 weighted images, hyperintense lesions on diffusion-weighted images, and results of follow-up CT or MR scans. RESULTS: In 58 (96.7%) patients, FLAIR detected intraarterial signals as early as 35 minutes after stroke onset. In 48 (80.0%) patients, intraarterial signal on FLAIR images coincided with lack of TOF on MR angiograms. In 41 (74.5%) of 55 patients, the intraarterial signals of fast T2 weighted imaging depicted occlusion better than did deficient flow void on T2 weighted images. In 25 (41.7%) of 60 patients, the area of intraarterial signal distribution was larger than the hyperintense lesion measured on diffusion weighted images. Areas of final infarction had sizes between those of intraarterial signal distribution on FLAIR images and lesions measured on diffusion-weighted images. In 35 (87.5%) of 40 patients, areas of intraarterial signal distribution were equal to regions of abnormal perfusion. CONCLUSION: Intraarterial signal on FLAIR images is an early sign of occlusion of major arteries. FLAIR combined with diffusion-weighted imaging can be helpful to predict an area at risk for infarction (ischemic penumbra). FLAIR plays an important role for determining whether a patient should undergo perfusion study. PMID- 11415893 TI - Diffusion and perfusion MR imaging in cases of Alzheimer's disease: correlations with cortical atrophy and lesion load. AB - BACKGROUND AND PURPOSE: Perfusion and diffusion-weighted MR imaging are powerful new imaging techniques for evaluating tissue pathophysiology in association with many neurologic disorders, such as neurodegenerative diseases. The purpose of our study was to evaluate the sensitivity and specificity of dynamic susceptibility contrast-enhanced MR imaging and diffusion-weighted MR imaging in cases of Alzheimer's disease and to assess the role of atrophy in the quantification of cortical perfusion. METHODS: Thirty-nine participants were studied: 18 patients with moderate cognitive impairment with probable Alzheimer's disease, 16 patients with mild impairment with possible or probable Alzheimer's disease, and 15 group matched elderly healthy comparison volunteers. Relative values of temporoparietal, sensorimotor, and hippocampal regional cerebral blood volume (rCBV) were measured as a percentage of cerebellar rCBV, and group classification was assessed with logistic regression. Brain atrophy was used as a covariate to assess its role in rCBV quantification. Regions of interest placed on orientation independent apparent diffusion coefficient maps allowed the calculation of apparent diffusion coefficient values and relative anisotropic indices of the head of the caudate nuclei, thalamus, parietal, frontal, and hippocampal cortices bilaterally, genu and splenium of corpus callosum, and anterior and posterior white matter in patients with Alzheimer's disease and in control volunteers. RESULTS: Temporoparietal rCBV ratios were reduced bilaterally in the patients with Alzheimer's disease. Sensitivity was 91% in moderately affected patients with Alzheimer's disease and 90% in patients with mild cases. Specificity was 87% in healthy comparison volunteers. Lower values of sensitivity and specificity were obtained for sensorimotor (73%, 50%, and 67%, respectively) and hippocampal cortices (80%, 80%, and 65%, respectively). Using brain atrophy as a covariate, patients with Alzheimer's disease still showed a statistically significant reduction of rCBV compared with control volunteers. Diffusion-weighted MR imaging analysis only showed a trend, with no statistic significance, of reduction of anisotropy in posterior white matter. CONCLUSION: Dynamic susceptibility contrast enhanced MR imaging of rCBV may be an alternative to nuclear medicine imaging for the evaluation of patients with Alzheimer's disease. When brain atrophy is used as a covariate, differences in rCBV still persist between patients with Alzheimer's disease and control volunteers, suggesting that perfusion impairment is unrelated to atrophy. No significant results for either white or gray matter were obtained using diffusion-weighted MR imaging. PMID- 11415894 TI - Microembolic signals and diffusion-weighted MR imaging abnormalities in acute ischemic stroke. AB - BACKGROUND AND PURPOSE: The clinical significance of microembolic signals (MESs) detected by transcranial Doppler sonography (TCD) in acute ischemic stroke remains unclear. The purpose of the present study was to assess the findings of diffusion-weighted MR imaging (DWI) and other clinical characteristics in patients with acute ischemic stroke and MESs. METHODS: We performed TCD and DWI within 48 hours and 7 days, respectively, after stroke onset in 28 patients with acute brain infarction. The relationship between the number of MESs and DWI findings, risk factors for stroke, National Institutes of Health Stroke Scale (NIHSS) score on admission, and arterial disease was examined. RESULTS: Ten patients had MESs detected by TCD (MES group) and 18 had no MESs (control group). The frequency of hypertension, diabetes mellitus, hyperlipidemia, and smoking; NIHSS score; blood-coagulation parameters; and interval between stroke onset and DWI study did not differ between the two groups. However, arterial disease was more frequent in the MES group than in the control group. Small, multifocal ischemic lesions (<10 mm in diameter) on DWI were more frequent in the MES group than in the control group. Conventional CT and MR imaging often failed to show these lesions. CONCLUSION: Small, often asymptomatic DWI abnormalities were more frequent in patients with MESs detected by TCD and with large-vessel occlusive diseases than in stroke patients without MESs. TCD and DWI may provide early clues to the mechanism of stroke in the acute phase. PMID- 11415895 TI - The role of functional MR imaging in patients with ischemia in the visual cortex. AB - BACKGROUND AND PURPOSE: Functional MR (fMR) imaging is based on changes in regional blood flow. The purpose of this study was to evaluate the role of fMR imaging for detection of a vascular compromised status in the occipital lobe in patients with ischemia in the visual cortex. METHODS: We performed fMR imaging in seven control subjects and seven patients with symptoms and signs of visual cortical transient ischemia and/or infarct. fMR imaging was performed using a gradient-echo sequence with the 2D fast low-angle shot technique. An axial slice including both visual cortices was selected, and stimulation of the visual cortex was performed using a red photostimulator. The number of activated pixels in each primary visual cortex area were counted and an asymmetry ratio [AR (%) = 100 x (R L)/(R+L)/2] was calculated. Patients and control subjects underwent visual field examination, conventional MR imaging, and vascular imaging (MR angiography in all patients and control subjects, conventional catheter angiography in two patients). fMR imaging results were compared with the results of a visual field examination, conventional MR imaging, and vascular imaging. RESULTS: fMR imaging of the patients showed significant activation asymmetry (P <.05) compared with that of control subjects. Vascular abnormalities in the posterior circulation were found in all seven patients. By conventional MR imaging, five patients were found to have infarction in the occipital lobe and the remaining two patients showed no abnormality. In visual field examination, six of the seven patients showed homonymous hemi- or quadrantanopsia suggesting postchiasmic abnormalities, and the remaining patient had normal findings. fMR imaging showed decreased activity in the visual cortices corresponding to vascular abnormalities (seven of seven patients), permanent infarction (five of seven patients), or visual field defect (six of seven patients). Two patients with normal conventional MR imaging had vascular lesions in the posterior circulation, and fMR imaging showed decreased activity in the corresponding visual cortices. One patient with normal visual field examination had multifocal stenosis in the posterior cerebral artery without infarction, and fMR imaging showed decreased activity in the corresponding visual cortex. CONCLUSION: fMR imaging of the visual cortex may be a sensitive method for the detection of vascular-compromised status in the occipital lobe. PMID- 11415896 TI - Automated CT segmentation and analysis for acute middle cerebral artery stroke. AB - BACKGROUND AND PURPOSE: The quantitative nature of CT should make it amenable to semiautomated analysis using modern neuroimaging methods. The purpose of this study was to begin to develop automated methods of analysis of CT scans to identify putative hypodensity within the lentiform nucleus and insula in patients with acute middle cerebral artery stroke. METHODS: Thirty-five CT scans were retrospectively selected from our CT archive (scans of 20 normal control participants and 15 patients presenting with acute middle cerebral artery stroke symptoms). The DICOM data for each participant were interpolated to a single volume, scalp stripped, normalized to a standard atlas, and segmented into anatomic regions. Voxel densities in the lentiform nucleus and insula were compared with the contralateral side at P <.01 using the Wilcoxon two-sample rank sum statistic, corrected for spatial autocorrelation. RESULTS: The quality of the registration for the anatomic regions was excellent. The control group had two false-positive results. The patient group had two false-negative results in the lentiform nucleus, two false-negative results in the insular cortex, and one false-positive finding for the insular cortex. The remainder of the infarcts were correctly identified. The original clinical reading, performed at the time of presentation, produced five false-negative interpretations for the patient group, all of which were correctly identified by the automated algorithm. CONCLUSION: We present an automated method for identifying potential areas of acute ischemia on CT scans. This approach can be extended to other brain regions and vascular territories and may aid in the interpretation of CT scans in cases of hyperacute stroke. PMID- 11415897 TI - Measurement of time-to-peak parameter by use of a new standardization method in patients with stenotic or occlusive disease of the carotid artery. AB - BACKGROUND AND PURPOSE: A quantitative, easily obtained measure of cerebral hemodynamics would be valuable in planning surgical or interventional therapy for patients with stenotic or occlusive disease of the carotid artery. We evaluated the recently introduced standardized time-to-peak variable (stdTTP), obtained with dynamic susceptibility contrast-enhanced MR (DSC-MR) imaging, in different states of stenotic/occlusive carotid artery disease. METHODS: We examined 24 patients with unilateral, high-grade stenosis (85%-95%) of one internal carotid artery (ICA) and 10 patients with stenosis (85%-95%) of one ICA and occlusion of the contralateral ICA. Mean stdTTP was evaluated in the central vascular territories of the anterior, middle, and posterior cerebral arteries and the anterior and posterior border zones and compared with the mean stdTTP values from 36 cerebral hemispheres without hemodynamic impairment. RESULTS: Patients showed no significant prolongation of stdTTP in the central vascular territories compared with the reference group, whereas significant prolongations of stdTTP were measured in the anterior and posterior border zones in patients with ICA disease (ANOVA, P <.05) and were most prominent in higher grades of carotid disease. Hemispheres with hemodynamic impairment always showed a stdTTP > 3.5 s in the border zones. CONCLUSION: The StdTTP quantitatively describes the hemodynamic impairment in cerebral hemispheres supplied by a stenosed or occluded ICA. An stdTTP value of > 3.5 s, as has been postulated, seems to point out hemodynamic impairment. PMID- 11415898 TI - Cerebral hemodynamics in asymptomatic patients with internal carotid artery occlusion: a dynamic susceptibility contrast MR and transcranial Doppler study. AB - BACKGROUND AND PURPOSE: Perfusion imaging with dynamic susceptibility contrast MR imaging (DSC-MRI) has been used to evaluate hemodynamic status in patients with symptomatic occlusive cerebrovascular disease. The aim of the present study was to determine the hemodynamic changes occurring in asymptomatic patients with unilateral internal carotid artery (ICA) occlusion by use of DSC-MRI with transcranial Doppler (TCD) measurement of the breath-holding index (BHI). METHODS: Nine patients with asymptomatic unilateral ICA occlusion underwent DSC MRI and TCD examination. One patient was excluded from final analysis because of severe movement artifacts. On a separate workstation, regional cerebral blood volume (rCBV), regional cerebral blood flow (rCBF), and regional mean time to peak (rMTT) were calculated on the basis of signal decay rate during the passage of gadolinium bolus through the sampled volume in the territory of the middle cerebral artery. TCD-BHI was calculated in all patients. Six healthy subjects underwent the same MR protocol as the patients. RESULTS: Compared with control subjects, patients with unilateral ICA occlusions had hemodynamic changes in the ipsilateral hemisphere: rCBF was significantly lower than in controls (P <.01), and r MTT was significantly increased in both white (WM) and gray matter (GM) in the affected side (WM: P <.01; GM: P <.05). No statistically significant difference in rCBV was found in the group of patients (occluded versus contralateral, P <.1) or between the patient and control groups (occluded side versus controls, P <.1). The correlation of rCBV and BHI showed a strong relation of the two variables, showing a decrease of the latter when the former increased. CONCLUSION: DSC-MRI is a valuable tool for measuring hemodynamic changes in the presence of carotid disease with hemodynamic impairment. In our opinion, hemodynamic changes and efficiency of collateral pathways can be evaluated in occlusive carotid disease by using paired measurement of BHI and DSC-MRI. In the patient group, MR-determined rCBV and TCD-determined BHI showed a significant inverse correlation, suggesting similar significance of the two indices. PMID- 11415899 TI - Diffusion-weighted imaging shows cytotoxic and vasogenic edema in eclampsia. AB - In eclampsia, MR imaging shows reversible T2 hyperintensities in a parietal and occipital distribution. Findings on diffusion-weighted images suggest that these abnormalities are areas of vasogenic edema. We describe the presence of both cytotoxic and vasogenic edema, as detected by diffusion-weighted imaging, in a woman with eclampsia. Follow-up MR imaging showed that the regions of cytotoxic edema progressed to cerebral infarction. This case suggests that diffusion weighted imaging allows the early detection of ischemic infarcts in patients with eclampsia. PMID- 11415900 TI - Cerebral amyloid angiopathy presenting as nonhemorrhagic diffuse encephalopathy: neuropathologic and neuroradiologic manifestations in one case. AB - A case of cerebral amyloid angiopathy is presented with MR imaging findings of high intense signal on T2-weighted sequences at the level of the white and gray matter of both hemispheres in the absence of neuroradiologic signs of cerebral hemorrhage. The biopsy specimen revealed deposition of amyloid in the walls of the intracranial arterial branches and focal ischemic changes and gliosis in the gray and white matter. We consider this presentation to be very unusual in patients affected by cerebral amyloid angiopathy. PMID- 11415901 TI - Multisection dynamic CT perfusion for acute cerebral ischemia: the "toggling table" technique. AB - A new CT perfusion technique providing extended anatomic coverage was evaluated in 12 patients with suspected acute middle cerebral artery ischemia. With a multidetector CT scanner, scans were obtained in an alternating fashion at two distinct "toggling" table positions (two 1-cm sections each) during a 40-mL contrast agent bolus (approximately 5 seconds per image), and perfusion parameter maps were created. The CT perfusion results were compared with follow-up images. Nine patients showed focal perfusion abnormalities in at least one section, most commonly on mean transit time maps. Using a single table location would have underestimated or missed the involved tissue in most cases. In three of 12 patients, perfusion maps failed to delineate any abnormality. In two patients, perfusion and diffusion MR imaging confirmed the absence of perfusion abnormality and tissue injury, respectively. In one case, a small ischemic injury was revealed by diffusion MR imaging. By using the toggling-table approach, perfusion images can be obtained over an extended anatomic area and, thus, reveal the presence and the extent of presumed tissue injury. PMID- 11415902 TI - The role of diffusion-weighted imaging in patients with brain tumors. AB - BACKGROUND AND PURPOSE: Diffusion-weighted images (DWIs) have been used to study various diseases, particularly since echo-planar techniques shorten examination time. Our hypothesis was that DWIs and tumor apparent diffusion coefficients (ADCs) could provide additional useful information in the diagnosis of patients with brain tumors. METHODS: Using a 1.5-T MR unit, we examined 56 patients with histologically verified or clinically diagnosed brain tumors (17 gliomas, 21 metastatic tumors, and 18 meningiomas). We determined ADC values and signal intensities on DWIs both in the solid portion of the tumor and in the peritumoral, hyperintense areas on T2-weighted images. We also evaluated the correlation between ADC values and tumor cellularity in both gliomas and meningiomas. RESULTS: The ADCs of low-grade (grade II) astrocytomas were significantly higher (P =.0004) than those of other tumors. Among astrocytic tumors, ADCs were higher in grade II astrocytomas (1.14 +/- 0.18) than in glioblastomas (0.82 +/- 0.13). ADCs and DWIs were not useful in determining the presence of peritumoral neoplastic cell infiltration. The ADC values correlated with tumor cellularity for both astrocytic tumors (r = -.77) and meningiomas (r = -.67). CONCLUSION: The ADC may predict the degree of malignancy of astrocytic tumors, although there is some overlap between ADCs of grade II astrocytomas and glioblastomas. PMID- 11415903 TI - Quantitative MR evaluation of intracranial epidermoid tumors by fast fluid attenuated inversion recovery imaging and echo-planar diffusion-weighted imaging. AB - BACKGROUND AND PURPOSE: Quantification of MR can provide objective, accurate criteria for evaluation of a given MR sequence. We quantitatively compared conventional MR sequences with fast fluid-attenuated inversion recovery (fast FLAIR) and echo-planar diffusion-weighted (DW) MR imaging in the examination of intracranial epidermoid tumors. METHODS: Eight patients with surgically confirmed intracranial epidermoid tumors were examined with T1-weighted MR sequences, fast T2- and proton density-weighted dual-echo sequences, fast-FLAIR sequences, and DW echo-planar sequences. We measured the MR signal intensity and apparent diffusion coefficient (ADC) of epidermoid tumors, normal brain tissue, and CSF and calculated the tumor-to-brain and tumor-to-CSF contrast ratios and contrast-to noise ratios (CNR). Results were compared among the five MR methods. RESULTS: On fast-FLAIR imaging, the mean signal intensity of epidermoid tumors was significantly higher than that of CSF but significantly lower than that of the brain; the contrast ratio and CNR of tumor-to-CSF were 4.71 and 9.17, respectively, significantly greater than the values with conventional MR imaging. On echo-planar DW imaging, epidermoid tumors showed a remarkably hyperintense signal relative to those of the brain and CSF; the mean contrast ratio and CNR of tumor-to-CSF were 13.25 and 19.34, respectively, significantly greater than those on fast-FLAIR or conventional MR imaging. The mean ADC of epidermoid tumors was 1.197 x 10(-3) mm(2)/s, significantly lower than that of CSF but higher than that of brain tissues. CONCLUSION: Fast-FLAIR imaging is superior to conventional MR imaging in depicting intracranial epidermoid tumors. Echo-planar DW imaging provides the best lesion conspicuity among the five MR methods. The hyperintensity of epidermoid tumors on echo-planar DW imaging is not caused by the diffusion restriction but by the T2 shine-through effect. PMID- 11415904 TI - Pituitary adenomas: early postoperative MR imaging after transsphenoidal resection. AB - BACKGROUND AND PURPOSE: Although there have been several reports on postoperative MR imaging of the sella, immediate postoperative changes (usually within 3 days) have not been extensively analyzed. The purpose of this study was to establish the value of early postoperative MR imaging in differentiating residual tumor from postoperative surgical changes in the sella after transsphenoidal resection of pituitary adenomas. METHODS: Eighty-three patients with surgically proven pituitary adenomas (32 nonfunctioning, 24 prolactin-secreting, 22 growth hormone secreting, and five prolactin- and growth hormone-secreting tumors) were studied prospectively. All patients underwent dynamic MR imaging within 7 days after surgery. We analyzed the postoperative MR images by focusing on changes in the pituitary gland, signal intensity, resorption of implanted material, and visibility of residual tumor. The patients were divided into four groups according to enhancement pattern of the postoperative pituitary mass: no enhancement, nodular enhancement, peripheral rim enhancement, and a combination of nodular and peripheral rim enhancement. RESULTS: Postoperative changes included resorption of implanted material and reexpansion of the pituitary gland. In 22 patients, residual tumors were found, and all patients showed nodular or combined enhancement. The residual tumors were confirmed by immediate reoperation in three patients, by hormonal assay and follow-up MR images in 11 patients with functioning adenomas, and by growth of the tumor on follow-up MR images in eight patients with nonfunctioning adenomas. Forty-eight patients showed no enhancement and 13 patients showed peripheral rim enhancement. CONCLUSION: Early postoperative dynamic MR imaging after transsphenoidal resection in pituitary adenoma is very effective in differentiating residual tumor from postoperative surgical changes. PMID- 11415905 TI - Parasagittal ependymoma resembling falcine meningioma. AB - Presentation, diagnosis, and management of an unusual parasagittal ependymoma, radiographically resembling a falcine meningioma, are described. Despite its radiographic appearance, pathologic evaluation revealed classic features of an ependymoma. The radiographic and pathologic characteristics of this unusual lesion are briefly examined, and the literature is reviewed. Although extraaxial ependymomas are rare, they should be considered in the radiographic differential diagnosis of dural-based lesions, especially for patients within the first 3 decades of life. PMID- 11415906 TI - Dynamic contrast-enhanced T2*-weighted MR imaging of tumefactive demyelinating lesions. AB - PURPOSE: Dynamic contrast-enhanced T2*-weighted MR imaging has been helpful in characterizing intracranial mass lesions by providing information on vascularity. Tumefactive demyelinating lesions (TDLs) can mimic intracranial neoplasms on conventional MR images, can be difficult to diagnose, and often result in surgical biopsy for suspected tumor. The purpose of this study was to determine whether dynamic contrast-enhanced T2*-weighted MR imaging can be used to distinguish between TDLs and intracranial neoplasms that share common features on conventional MR images. METHODS: We retrospectively reviewed the conventional and dynamic contrast-enhanced T2*-weighted MR images and medical records of 10 patients with tumefactive demyelinating disease that was diagnosed by either biopsy or strong clinical suspicion supported by laboratory evaluation that included CSF analysis and evoked potential tests. Twelve TDLs in 10 patients and 11 brain tumors that appeared similar on conventional MR images were studied. Relative cerebral blood volume (rCBV) was calculated from dynamic MR data and was expressed as a ratio to contralateral normal white matter. rCBV values from 11 patients with intracranial neoplasms with very similar conventional MR imaging features were used for comparison. RESULTS: The rCBV values of TDLs ranged from 0.22 to 1.79 (n = 12), with a mean of 0.88 +/- 0.46 (SD). The rCBV values of intracranial neoplasms ranged from 1.55 to 19.20 (n = 11), with a mean of 6.47 +/ 6.52. The difference in rCBV values between the two groups was statistically significant (P =.009). The difference in rCBV values between TDLs and primary cerebral lymphomas (n = 4) was less pronounced but was statistically significant (P =.005). CONCLUSION: Dynamic contrast-enhanced T2*-weighted MR imaging is a useful diagnostic tool in differentiating TDLs from intracranial neoplasms and may therefore obviate unnecessary surgical biopsy. PMID- 11415907 TI - Delayed MR imaging changes in acute disseminated encephalomyelitis. AB - BACKGROUND AND PURPOSE: White matter lesions on MR images obtained from patients with acute disseminated encephalomyelitis (ADEM) have been reported to appear shortly after symptom onset, and their resolution has been claimed to parallel recovery. To elucidate the temporal evolution of these lesions and to associate the changes on MR images to the patients' clinical condition, we performed serial MR imaging on patients with ADEM. METHODS: Several consecutive T2-weighted and fluid-attenuated inversion recovery scans were obtained from four previously healthy adult patients with ADEM within the first days after the onset of symptoms and again during the recovery period. MR imaging was done first on a weekly to biweekly basis and later at 1- to 2-month intervals for up to 8 months. RESULTS: MR scans of three of these patients did not show any specific abnormalities until several weeks after the onset of the disease. As the lesions later appeared, their number increased during the recovery period. CONCLUSION: MR imaging performed during the first days after the onset of the disease may not reveal any pathologic findings. The appearance of the ADEM-associated MR imaging changes may be associated with recovery rather than decline. It remains to be studied whether the new MR imaging techniques reveal the lesions associated with ADEM faster than the conventional T2-weighted imaging. PMID- 11415908 TI - Quantitative proton MR spectroscopic imaging in acute disseminated encephalomyelitis. AB - Serial MR imaging and quantitative proton MR spectroscopic imaging (MRSI) findings of a 4-year-old boy with acute disseminated encephalomyelitis (ADEM) are reported. Over a 2-month period characterized by an initial illness and two relapses, each with full recovery, MR imaging exhibited the appearance and disappearance of multifocal lesions throughout the CNS that correlated only partly with the neurologic impairment. During one relapse, MRSI revealed low levels of N-acetylaspartate (NAA) within the regions of prolonged T2 signal intensity. All other metabolites were normal. At follow-up, the MR imaging and MRSI abnormalities had fully resolved. MRSI might play an important role in the diagnosis of ADEM, as well as in the elucidation of underlying pathophysiologic processes in this poorly defined disorder of children. This case demonstrates that reduced levels of NAA are not always associated with neuronal loss, irreversible tissue damage, or poor neurologic outcome. PMID- 11415909 TI - Coexistent neurocysticercosis and Japanese B encephalitis: MR imaging correlation. AB - BACKGROUND AND PURPOSE: An increased incidence of intestinal helminthic infections has been observed in patients with viral encephalitis in endemic areas. Both Japanese B encephalitis (JE) and neurocysticercosis (NCC) share some common socio-demographic and ecologic factors, and pigs act as the intermediate carrier for both. Our purpose was to show the coexistence of JE and NCC in brain on MR images and highlight the possible role of NCC as an amplifier of JE. METHODS: MR images from 10 cases of coexistent JE and NCC were studied retrospectively. T1-weighted axial and sagittal, proton T2-weighted axial and coronal, and T2-weighted fluid-attenuated inversion recovery axial and coronal sections of the brain were evaluated. NCC was diagnosed on the basis of neuroimaging. Diagnostic serologic testing for JE was conducted using paired blood and CSF samples. RESULTS: The JE changes were bilateral and asymmetrical and were more severe on the side harboring the solitary cyst or the side bearing the greater number of cysts or lodging the degenerating cyst. In each of nine of 10 cases, at least one degenerating cyst was found on the side of predominant JE pathologic abnormality. CONCLUSION: The study suggests that the co-occurrence of JE and NCC is not just a chance coincidence. NCC apparently predisposes a person to JE infection and is a positive modulator of the encephalitic process. The study shows a spectrum of MR imaging findings of coexistent JE and NCC. PMID- 11415910 TI - Magnetization transfer ratio values and proton MR spectroscopy of normal appearing cerebral white matter in patients with liver cirrhosis. AB - BACKGROUND AND PURPOSE: Hepatic encephalopathy in cirrhotic patients may be the clinical manifestation of disturbed cerebral cell volume homeostasis. The aim of this study was to investigate the presence of significant changes in magnetization transfer ratio (MTR) values, which could reflect an increase in free water within the brain of patients with liver cirrhosis, and to correlate these findings with minimal hepatic encephalopathy and proton MR spectroscopy ((1)H-MRS) abnormalities. METHODS: Twenty-four patients with liver cirrhosis and eight healthy control volunteers were included in the study. MR imaging studies included conventional T1- and T2-weighted imaging, (1)H-MRS, and magnetization transfer imaging. MTR and (1)H-MRS values were obtained from normal-appearing white matter and were correlated with each other and with the presence of minimal hepatic encephalopathy. RESULTS: (1)H-MRS showed a decrease in choline and myo inositol and an increase in glutamine + glutamate with respect to creatine + phosphocreatine. MTR values were significantly decreased in cirrhotic patients when compared with healthy control volunteers, although this decrease was not significantly higher in the patients with minimal hepatic encephalopathy. The decreases in MTR values correlated with increases in glutamine + glutamate. CONCLUSION: The MTR decrease in patients with liver cirrhosis may be caused by low-grade astrocytic swelling produced as a response to the osmotic stress occurring in these patients. However, in this cross-sectional study, we did not find a correlation between MTR decrease and the presence of minimal hepatic encephalopathy, probably because of limitations in MTR quantification techniques. PMID- 11415911 TI - MR imaging after aneurysmal subarachnoid hemorrhage and surgery: a long-term follow-up study. AB - BACKGROUND AND PURPOSE: We assumed that patients with surgically treated aneurysmal subarachnoid hemorrhage (SAH) might have more lesions than those revealed by CT that could be visible on MR images. METHODS: We conducted a retrospective study of a series of 147 patients with aneurysmal SAH who were treated surgically within 3 days of the onset of SAH. One hundred four patients (mean age, 48.8 years) underwent MR imaging studies 2.1 to 5.6 years (mean, 3.3 years) postoperatively. RESULTS: Eighty-four (81%) patients presented a total of 152 areas of increased signal intensity on T2-weighted images, consistent with infarction; 48% of the patients had lesions in the frontal lobes. CT performed 3 months postoperatively revealed hypodense areas on the scans of only 57% of the patients and showed lesions in the frontal lobes of only 16% of the patients. CONCLUSION: Patients who undergo early surgery for aneurysmal SAH have more lesions than are revealed by CT. The difference is remarkable, especially in the frontal lobes. PMID- 11415912 TI - Transient MR signal changes in patients with generalized tonicoclonic seizure or status epilepticus: periictal diffusion-weighted imaging. AB - BACKGROUND AND PURPOSE: Our purpose was to investigate transient MR signal changes on periictal MR images of patients with generalized tonicoclonic seizure or status epilepticus and to evaluate the clinical significance of these findings for differential diagnosis and understanding of the pathophysiology of seizure induced brain changes. METHODS: Eight patients with MR images that were obtained within 3 days after the onset of generalized tonicoclonic seizure or status epilepticus and that showed seizure-related MR signal changes had their records retrospectively reviewed. T1- and T2-weighted images were obtained of all eight patients. Additional diffusion-weighted images were obtained of five patients during initial examination. After adequate control of the seizure was achieved, follow-up MR imaging was performed. We evaluated the signal changes, location of the lesions, and degree of contrast enhancement on T1- and T2-weighted images and the signal change and apparent diffusion coefficient (ADC) on diffusion-weighted images. We also compared the signal changes of the initial MR images to those of the follow-up MR images. RESULTS: The initial MR images revealed focally increased T2 signal intensity, swelling, and increased volume of the involved cortical gyrus in all eight patients. The lesions were located in the cortical gray matter or subcortical white matter in seven patients and at the right hippocampus in one. T1-weighted images showed decreased signal intensity at exactly the same location (n = 6) and gyral contrast enhancement (n = 4). Diffusion-weighted images revealed increased signal intensity at the same location and focally reduced ADC. The ADC values were reduced by 6% to 28% compared with either the normal structure opposite the lesion or normal control. Follow-up MR imaging revealed the complete resolution of the abnormal T2 signal change and swelling in five patients, whereas resolution of the swelling with residual increased T2 signal intensity at the ipsilateral hippocampus was observed in the other two patients. For one of the two patients, hippocampal sclerosis was diagnosed. For the remaining one patient, newly developed increased T2 signal intensity was shown. CONCLUSION: The MR signal changes that occur after generalized tonicoclonic seizure or status epilepticus are transient increase of signal intensity and swelling at the cortical gray matter, subcortical white matter, or hippocampus on periictal T2-weighted and diffusion-weighted images. These findings reflect transient cytotoxic and vasogenic edema induced by seizure. The reversibility and typical location of lesions can help exclude the epileptogenic structural lesions. PMID- 11415913 TI - Age and sex differences in the cerebellum and the ventral pons: a prospective MR study of healthy adults. AB - BACKGROUND AND PURPOSE: The human brain exhibits a complex pattern of differential aging. The purpose of this study was to examine whether age differences in the volume of cerebellar regions and the ventral pons are differential or generalized, whether the age-related shrinkage is linear or exponential, and whether there are sex differences in the size of the cerebellum and pons. METHODS: The volumes of the cerebellar hemispheres (excluding the vermis and the peduncles), the vermis, and the ventral pons were estimated from the prospectively acquired MR scans of 190 healthy volunteers (aged 18-81 years). The relation between regional volumes, age, and sex was assessed while taking into account differences in body size (height). RESULTS: We found a moderate age related reduction in the volume of the cerebellar hemispheres and the cerebellar vermis. In contrast to previous findings that suggested differential vulnerability of the posterior vermis, the age-related shrinkage of the vermian lobules was uniform-about 2% per decade. In accord with all reports in the literature, the size of the ventral pons was unrelated to age. The volume of the cerebellar hemispheres, the vermis, and the ventral pons were larger in men, even after adjustment for height. The magnitude of the sex difference was the largest in the hemispheres and the anterior vermis, and the smallest in the lobules VI VII (declive-folium-tuber). CONCLUSION: Moderate age-related shrinkage of the cerebellum and lack of age-related differences in the ventral pons are robust phenomena. However, in all likelihood, the effects of age on the cerebellum are not differential but uniform. The cerebellum and the pons are larger in men than in women and the difference is especially pronounced in the cerebellar hemispheres and the anterior vermis. PMID- 11415914 TI - Recurrent neurovascular hypertension: MR findings before and after surgical treatments. AB - A case of recurrent brain stem edema after surgical vascular decompression in a patient with neurogenic hypertension is presented. The surgical treatment resulted in occlusion of the left vertebral artery, stable blood pressure values, and no recurrence of the brain stem edema. MR imaging and MR angiography are excellent methods with which to assess patients suspected of having neurovascular hypertension, both before and after surgical treatment. PMID- 11415915 TI - Postoperative complications in otospongiosis: usefulness of MR imaging. AB - BACKGROUND AND PURPOSE: Sensorineural hearing loss (SNHL) is a rare complication of stapes surgery that may arise for many reasons. Usually, the pathogenesis of SNHL can be established by clinical and CT examinations. The purpose of this study was to evaluate the utility of MR imaging when CT findings are normal or not contributive. METHODS: Eleven patients with SNHL (in some instances, associated with vertigo) after stapedectomy, in whom CT showed no well-defined cause, were examined by MR imaging. RESULTS: MR studies established the additional findings of reparative intravestibular granuloma (n = 2), intralabyrinthine hemorrhage (n = 1), and bacterial labyrinthitis (n = 1). In five cases, MR findings were similar to CT findings. In two cases, CT and MR results were normal. Revision surgery was performed in five patients and confirmed the MR findings in each case. CONCLUSION: If CT is not contributive as to the origin of SNHL and vertigo occurring after stapes surgery, then MR imaging may be helpful in these patients. PMID- 11415916 TI - MR cisternography of the cerebellopontine angle: comparison of three-dimensional fast asymmetrical spin-echo and three-dimensional constructive interference in the steady-state sequences. AB - BACKGROUND AND PURPOSE: MR cisternography has been used as the noninvasive screening tool of the cerebellopontine angle. The purpose of this study was to directly compare two currently dominant types of sequences for heavily T2 weighted MR cisternography. METHODS: Three-dimensional fast asymmetric spin-echo (3D-FASE) sequences, which are 3D half-Fourier rapid acquisition with relaxation enhancement and 3D constructive interference in the steady-state (3D-CISS) sequences, were compared on a clinical 1.5-T MR unit using the same scan times. In five healthy volunteers, the contrast-to-noise ratio (C/N) between CSF and the cerebellum was measured at three locations. Then, for qualitative analysis, the quality of the labyrinth was scored on the original source multiplanar reformatted images, the virtual endoscopic images, and the maximum intensity projection (MIP) images. In 20 consecutive patients with suspected cerebellopontine angle tumors, visualization of the tumors was evaluated using 3D contrast-enhanced spoiled gradient-echo imaging as the standard of reference. RESULTS: Both sequences showed comparable mean C/N values; however, in qualitative analysis, the scores for 3D-CISS on the source, virtual endoscopic, and MIP images were significantly lower than those on the images obtained with 3D FASE, owing to more prominent flow and magnetic susceptibility artifacts on the 3D-CISS sequences. In all subjects, discontinuity of the semicircular canals was seen on the virtual endoscopic and MIP images obtained with 3D-CISS, owing to susceptibility artifacts, but not on those obtained with 3D-FASE. All 12 tumors were detected by both sequences, but 3D-CISS gave one false-positive result. CONCLUSION: 3D-FASE is considered the method of choice because artifacts are reduced and specificity is increased. PMID- 11415917 TI - The petrosquamosal sinus: CT and MR findings of a rare emissary vein. AB - BACKGROUND AND PURPOSE: Morphologic changes in the dural sinuses and emissary veins of the posterior fossa relate closely to the development of the brain. We report characteristic findings of imaging in six patients with a rare and forgotten emissary vein called the petrosquamosal sinus (PSS). METHODS: From a larger group of patients with ear abnormalities, we selected six patients from three ENT imaging centers, because they had CT features suggestive of a PPS. This was the criterion for inclusion in this retrospective study. They were explored by high-resolution CT (HRCT) of the temporal bone. MR venography was performed in three patients to determine the presence and patency of the emissary vein. RESULTS: The PPS was bilateral in two patients and unilateral in the other four. It affected mainly the left side (left:right ratio, 5:3). Three patients had associated inner ear (n = 2) or middle ear malformations (n = 1). Five of six patients had jugular vein hypoplasia, with development of emissary mastoid veins in three patients. CONCLUSION: Petrosquamosal sinus can be identified on HRCT in a typical location. It is encountered more frequently in patients referred for congenital abnormalities of the skull base. This rare anatomic variant should be assessed before surgical treatment, because proper identification of these large venous channels would be of interest to the surgeon. PMID- 11415918 TI - Radioprotection to the eye during CT scanning. AB - BACKGROUND AND PURPOSE: The lens of the eye is sensitive to radiation. Children undergoing CT of the head and patients undergoing repeated CT scanning of the head are vulnerable to this complication. The purpose of this study was to test the ability of a heavy metal, bismuth, in reducing radiation to the lens of the eye during routine cranial CT. METHODS: Both phantom and human studies were done. Using a standard head-attenuating phantom, scanning was performed with detectors placed over the eye, first without the protectors, and then with shielding by one (1T), two (2T), or three thickness (3T) of bismuth-coated latex. The patient study included 30 patients randomized into one of three groups with eye protection provided by 1T, 2T, or 3T of the bismuth-coated latex. Control measurements were done using thermoluminescent dosimeters over the forehead above each eye. Image artifact from the bismuth shields was assessed. RESULTS: The phantom study demonstrated that the use of bismuth-coated shielding over the eyes decreased radiation dosage by 48.5%, 59.8%, and 65.4% using 1T, 2T, and 3T, respectively. The effect of eye shielding in decreasing radiation dosage to the eye was highly significant for all three thicknesses (P = 2.9 x 10(-81) to 1.9 x 10(-89)). In the patient study, the use of 1T, 2T, and 3T of bismuth-coated latex saved an average radiation dose of 39.6%, 43.5%, and 52.8%, respectively. While the use of shielding was statistically significant in saving radiation for all thicknesses (P = 2.2 x 10(-10) to 1.4 x 10(-21)), there was no statistical difference between 1T, 2T, and 3T of bismuth-coated latex shielding found in patients. However, the trend was for increased radiation savings to the eye with increased thickness of shielding used. A review of all 30 studies showed no significant artifact caused by the eye shielding, regardless of thickness. CONCLUSION: Bismuth-coated latex shielding of the eye during cranial CT is simple to apply, inexpensive, and causes up to a 50% reduction in radiation to the lens of the eye. PMID- 11415919 TI - Imaging of hydrogel episcleral buckle fragmentation as a late complication after retinal reattachment surgery. AB - Hydrogel encircling bands were introduced in the early 1980s as a product that was superior to bands composed of silicone rubber or silicone sponge for the surgical treatment of retinal detachment. Late complications consisting of orbital swelling and diplopia requiring band removal began to be reported in the early 1990s. Pathologic studies of these expanded fragments of hydrogel material after removal showed in vivo hydrolysis with foreign body reaction and dystrophic calcification. We report the imaging findings in five patients in whom this late complication developed. Hydrogel fragmentation has a characteristic imaging appearance consisting of a circumferential orbital mass associated with rim enhancement. This appearance should prompt inquiries regarding previous scleral buckle procedures with hydrogel bands. Familiarity with this appearance will avoid misinterpretation and unwarranted biopsy before band removal. PMID- 11415920 TI - Synovial chondromatosis in the temporomandibular joint complicated by displacement and calcification of the articular disk: report of two cases. AB - Two cases of synovial chondromatosis of the temporomandibular joint (TMJ) are presented, including correlation of CT and MR imaging characteristics with surgical and pathologic findings. The usefulness of CT and MR imaging in the diagnosis of TMJ disorders is discussed. PMID- 11415921 TI - Perineural spread in a case of sinonasal sarcoidosis: case report. AB - We report a case of sinonasal sarcoidosis with perineural spread along the trigeminal and vidian nerves in which primarily MR imaging but also CT allowed excellent visualization of this infiltration. PMID- 11415922 TI - Spontaneous retropharyngeal hematoma: diagnosis by mr imaging. AB - Spontaneous retropharyngeal hematoma is an uncommon entity that is difficult to diagnose and may progress rapidly to airway obstruction. We report a case of a 53 year-old man with acute onset of retropharyngeal pain, dysphonia, and dysphagia after vomiting. On CT, a nonspecific retropharyngeal collection was seen. MR imaging demonstrated blood products, suggesting a diagnosis of retropharyngeal hematoma, and the patient was managed conservatively. MR imaging allowed specific diagnosis of a rare condition that is otherwise difficult to diagnose without surgical intervention. PMID- 11415923 TI - An ex vivo biomechanical evaluation of a hydroxyapatite cement for use with kyphoplasty. AB - BACKGROUND AND PURPOSE: Previous ex vivo biomechanical studies have shown that kyphoplasty with polymethylmethacrylate cement increases vertebral body (VB) strength and restores VB stiffness and height after compression fracture. The purpose of the current study was to determine if a hydroxyapatite cement used as a void filler during kyphoplasty provides mechanical stabilization similar to that of a polymethylmethacrylate cement. METHODS: Simulated compression fractures were experimentally created in 33 osteoporotic VBs harvested from female cadaver spines. VBs were assigned to one of three groups: 1) kyphoplasty with a custom mixture of Simplex P; 2) kyphoplasty with BoneSource; and 3) no treatment. The kyphoplasty treatment consisted of inserting a balloon-like device into the VB via both pedicles, inflating the tamp, and filling the created void with Simplex P bone cement or BoneSource. VBs in the no-treatment group received no interventions. Pre- and posttreatment heights were measured, and the repaired VBs were recompressed to determine posttreatment strength and stiffness values. RESULTS: Kyphoplasty with altered Simplex P restored strength, whereas kyphoplasty with BoneSource and the no-treatment protocol both resulted in significantly weaker VBs relative to initial strength. All treatments resulted in significantly less stiff VBs relative to their initial condition. All VBs lost significant height after initial compression, but a significant amount of lost height was restored by kyphoplasty with either cement. CONCLUSION: Kyphoplasty with either cement significantly restored VB height. Kyphoplasty with altered Simplex P resulted in stronger repairs than did no treatment or kyphoplasty with BoneSource. PMID- 11415924 TI - Dural arteriovenous fistula in children: endovascular treatment and outcomes in seven cases. AB - BACKGROUND AND PURPOSE: Dural AVF is a vascular anomaly that rarely occurs in children and is best treated by endovascular embolization. We report our experience using various endovascular embolization techniques in the treatment of dural AVF in a pediatric population. METHODS: Seven children with angiographically proven dural AVF were treated with endovascular embolization using microcoils, N-butylcyanoacrylate, detachable balloons, and/or silk suture. All imaging studies, embolization procedures, and patient charts were retrospectively reviewed. RESULTS: Seven children had been treated for dural AVF at our institution since 1987. Three newborns presented with congestive heart failure. Four older children (10 months-10 years) presented with signs referable to venous hypertension, including seizures, hydrocephalus, and proptosis. Embolization approaches included transarterial, transvenous, and direct puncture after neurosurgical exposure of a dural sinus. The number of embolizations ranged from 1 to 13 sessions per patient. All patients experienced symptomatic improvement after each embolization session. The three newborns showed marked improvement in cardiac function that allowed discharge to home. Clinical follow up ranged from 3 weeks to 9 years (mean, 4.1 years). Two children with partially embolized dural AVF died, and one was lost to follow-up. Four children are alive after complete embolization of their dural AVF; two are developmentally normal, and two have mild developmental delay. CONCLUSION: Endovascular embolotherapy is the current treatment of choice for dural AVF. Embolization therapy may be life saving in the setting of cardiac failure and curative in cases of small or simple fistulae. Multiple, complex dural AVF are usually not curable, and treatment is aimed at symptomatic relief. Treatment strategies focus on the location and/or complexity of the fistula, the patient's clinical status, and the neurologic prognosis. PMID- 11415927 TI - Stem cell niches in the mouse airway. PMID- 11415928 TI - Eotaxins. Contributing to the diversity of eosinophil recruitment and activation. PMID- 11415929 TI - Novel antiinflammatory targets for asthma. A role for PPARgamma? PMID- 11415930 TI - Evidence for stem-cell niches in the tracheal epithelium. AB - It is generally important to elucidate airway epithelial cell lineages and to identify multipotent progenitors as targets for gene therapy. Stem (S) cells are typically present in specialized compartments spatially proximal to their differentiated progeny, but an equivalent paradigm has not been demonstrated in the airway. We discovered a distinct population of cells displaying high levels of keratin expression in murine tracheal submucosal gland ducts, and tested the hypothesis that bromodeoxyuridine (BrdU) label-retaining cells (LRCs), thought to represent the S-cells, were present in this compartment. Mice received weekly epithelial damage by intratracheal detergent or SO(2) inhalation for 4 wk and received intraperitoneal injections of BrdU every 48 h during the injury and repair period. At 3 and 6 d after injury, BrdU-positive epithelial cells were noted along the entire tracheal length in both basal and lumenal cell positions. At later time points (20 and 95 d) LRCs were localized to gland ducts in the upper trachea and to systematically arrayed foci in the lower trachea, typically near the cartilage-intercartilage junction. LRCs were not pulmonary neuroendocrine cells. Heterotopic tracheal grafts after surface epithelial removal demonstrated reconstitution of a surface-like epithelium from gland remnants. These results suggest that airway epithelial S cells are localized to specific niches. PMID- 11415931 TI - Clara cell secretory protein-expressing cells of the airway neuroepithelial body microenvironment include a label-retaining subset and are critical for epithelial renewal after progenitor cell depletion. AB - Stem cells with potential to contribute to the re-establishment of the normal bronchiolar epithelium have not been definitively demonstrated. We previously established that neuroepithelial bodies (NEBs) sequester regenerative cells that contribute to bronchiolar regeneration after selective chemical depletion of Clara cells, a major progenitor cell population. Two candidate stem cells were identified on the basis of proliferative potential after chemical ablation: a pollutant-resistant subpopulation of Clara cells that retain their expression of Clara cell secretory protein (CCSP) (variant CCSP-expressing [CE] cells or vCE cells) and calcitonin gene-related peptide (CGRP)-expressing pulmonary neuroendocrine cells (PNECs). In the present study, two populations of label retaining cells were identified within the NEB: CGRP-expressing cells and a subpopulation of CE cells. To investigate contributions made by CE and CGRP expressing cells to epithelial renewal, CE cells were ablated through acute administration of ganciclovir to transgenic mice expressing herpes simplex virus thymidine kinase under the regulatory control of the mouse CCSP promoter. CGRP immunoreactive PNECs proliferated after depletion of CE cells, yet were unable to repopulate CE cell-depleted airways. These results support the notion that vCE cells represent either an airway stem cell or are critical for stem cell maintenance, and suggest that PNECs are not sufficient for epithelial renewal. PMID- 11415932 TI - Eotaxin-3 but not eotaxin gene expression is upregulated in asthmatics 24 hours after allergen challenge. AB - Eotaxin is an important mediator of eosinophil recruitment and activation in the airways of asthmatics. Eotaxin-2 and eotaxin-3 are two recently identified chemokines with activity similar to that of eotaxin. Using quantitative polymerase chain reaction analysis, we determined the messenger RNA (mRNA) expression of eotaxin, eotaxin-2, and eotaxin-3 relative to GAPDH mRNA expression in bronchial biopsies and bronchoalveolar lavage fluid (BALF) cells obtained from subjects with mild asthma, asthmatic subjects 24 h after allergen challenge, and normal control subjects. In bronchial biopsies, gene expression was upregulated in asthmatic subjects as compared with control subjects for eotaxin (log median values 3.18 pg/microg, 95% confidence interval [CI]; 2.27 to 3.79 versus 4.37 pg/microg, 95% CI; 3.97 to 4.65, P = 0.003) and for eotaxin-2 (0.82 pg/microg, 95% CI; 0.08 to 1.72 versus 2.97 pg/microg, 95% CI; 1.97 to 3.45, P = 0.006), but no further increase was observed after allergen challenge. In contrast, eotaxin-3 mRNA expression was not increased in asthmatic compared with control subjects, but was dramatically enhanced 24 h after challenge (median log value 1.93 pg/microg, 95% CI; 0.74 to 3.92 versus 4.62 pg/microg, 95% CI; 3.05 to 6.23, P = 0.036). No significant difference between groups was observed in BALF cell gene expression for any of the chemokines examined. These data suggest that eotaxin-3 rather than eotaxin or eotaxin-2 may account for the ongoing eosinophil recruitment to asthmatic airways in the later stages (24 h) following allergen challenge. PMID- 11415933 TI - Peroxisome proliferator-activated receptor-gamma regulates airway epithelial cell activation. AB - The peroxisome proliferator-activated receptors (PPARs) are nuclear hormone transcription factors that regulate genes associated with lipid and glucose metabolism. Recent evidence suggests that PPAR-gamma may also act as a negative immunomodulator. To investigate the potential role of PPAR-gamma in regulating airway inflammation, we characterized the expression and function of PPAR-gamma in airway epithelial cells. Airway epithelial cells constitutively express PPAR gamma-specific messenger RNA and protein. Further, airway epithelial PPAR-gamma is inducible by interleukin (IL)-4 in NIH-A549 cells. Two PPAR-gamma agonists, the prostaglandin D2 metabolite 15-deoxy-(Delta)(12,14) prostaglandin J2 (15d PGJ2) and a thiazolidinedione, ciglitazone, were used to study the effects of PPAR-gamma activation on airway epithelial cytokine expression. Activation of PPAR-gamma stimulated a PPAR-responsive reporter gene in a ligand-specific manner. In NIH-A549 cells, both ligands also blocked the cytokine-induced expression of the inducible form of nitric oxide synthase in a dose-dependent manner. In contrast, ciglitazone alone had a slight effect on cytokine-induced IL 8 secretion, but markedly inhibited IL-8 secretion from cells pretreated with IL 4. The demonstration of PPAR-gamma expression and function in airway epithelial cells expands the immunoregulatory role of PPARs and suggests a critical role for PPAR-gamma in antagonizing proinflammatory pathways in the airways. PMID- 11415934 TI - Paradoxical effects of platelet-derived growth factor-A overexpression in malignant mesothelioma. Antiproliferative effects in vitro and tumorigenic stimulation in vivo. AB - Malignant mesothelioma is associated with asbestos exposure and remains resistant to all therapeutic intervention. Previous studies have suggested an enhancing role for platelet-derived growth factor (PDGF) in mesothelial tumorigenicity, although the mechanism by which PDGF facilitates tumorigenicity is unknown. Here, we evaluate the contribution of PDGF-A expression to mesothelial tumorigenicity using ectopic modulation of PDGF-A expression. We find, in accordance with other reports, that the receptor for PDGF-A, although expressed at high levels in normal human mesothelial cells, is not easily detectable in mesothelioma. Further, we show that PDGF-A overexpression is responsible for autocrine downregulation of its receptor. Our data indicate, surprisingly, that for mesothelioma cells in vitro, high-level activation of a PDGF-A-PDGF receptor loop is antiproliferative whereas abrogation of PDGF-A expression stimulates growth. These data suggest that PDGF-A does not contribute to tumorigenicity by autocrine stimulation of proliferation. In contrast, increased PDGF-A expression in vivo increases tumor incidence and growth rate and decreases the latency period to tumor formation whereas abrogation of PDGF-A expression decreases tumor incidence and increases latency. Thus, the tumorigenic effect of PDGF-A must act through paracrine mechanisms relevant at early stages of tumor initiation. PMID- 11415935 TI - The cyclin-dependent kinase inhibitor p21 protects the lung from oxidative stress. AB - The lung is a major target tissue for oxidative stress, including hyperoxia used to relieve tissue hypoxia. Unfortunately, severe hyperoxia damages DNA, inhibits proliferation, and kills cells, resulting in morbidity and mortality. Although hyperoxia induces the tumor suppressor p53 and its downstream target, the cyclin dependent kinase inhibitor p21(Cip1/WAF1/Sdi1) (p21), their role in pulmonary injury remains unknown. Using p53- and p21-deficient mice we demonstrate that hyperoxia induces p21 in the absence of p53, suggesting that previous conclusions that p53 does not modify hyperoxic lung injury cannot be extrapolated to p21. In fact, mean survival of p21-deficient mice decreased by 40% and was associated with terminal deoxyribonucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick-end labeling staining of alveolar debris, indicative of DNA fragmentation and cell death. Ultrastructural analyses revealed that alveolar endothelial and type I epithelial cells died rapidly by necrosis. Although hyperoxia decreased DNA replication in p21-wild-type lungs, it had no effect on replication in p21-deficient lungs. Our findings suggest that p21 protects the lung from oxidative stress, in part, by inhibiting DNA replication and thereby allowing additional time to repair damaged DNA. Our findings have implications for patients suffering from the toxic effects of supplemental oxygen therapies. PMID- 11415936 TI - Differential regulation of sphingosine-1-phosphate- and VEGF-induced endothelial cell chemotaxis. Involvement of G(ialpha2)-linked Rho kinase activity. AB - We compared stimulus-coupling pathways involved in bovine pulmonary artery (PA) and lung microvascular endothelial cell migration evoked by sphingosine-1 phosphate (S1P), a potent bioactive lipid released from activated platelets, and by vascular endothelial growth factor (VEGF), a well-recognized angiogenic factor. S1P-induced endothelial cell migration was maximum at 1 microM (approximately 8-fold increase with PA endothelium) and surpassed the maximal response evoked by either VEGF (10 ng/ml) (approximately 2.5-fold increase) or hepatocyte growth factor (HGF) (approximately 2.5-fold increase). Migration induced by S1P, but not by VEGF, was significantly inhibited by treatment with antisense oligonucleotides directed to Edg-1 and Edg-3 (endothelial differentiation gene) S1P receptors and by G protein modification. These strategies included pretreatment with pertussis toxin, or transfection with mini genes encoding a betagamma subunit inhibitory peptide of the beta-adrenergic receptor kinase, or an 11-amino-acid peptide that inhibits G(1alpha2) signaling. Various strategies to interrupt Rho family signaling, including C(3) exotoxin, dominant/negative Rho, or the addition of Y27632, a cell-permeable Rho kinase inhibitor, significantly attenuated S1P- but not VEGF-induced migration. Conversely, pharmacologic inhibition of either myosin light chain kinase, src family tyrosine kinases, or phosphatidylinositol-3' kinase reduced basal endothelial cell migration and abolished VEGF-induced endothelial cell migration but did not inhibit the increase in S1P-induced migration. Whereas VEGF and S1P increased both p42/p44 extracellular regulated kinase and p38 mitogen-activated protein (MAP) kinase activities, only p38 MAP kinase inhibition significantly reduced VEGF- and S1P-stimulated migration. These data confirm S1P as a potent endothelial cell chemoattractant through G(1alpha2)-coupled Edg receptors linked to Rho-associated kinase and p38 MAP kinase activation. The divergence in signaling pathways evoked by S1P and VEGF suggests complex and agonist-specific regulation of endothelial cell angiogenic responses. PMID- 11415937 TI - Double-stranded rna dependence of nitric oxide synthase 2 expression in human bronchial epithelial cell lines BET-1A and BEAS-2B. AB - The human airway epithelium expresses abundant nitric oxide synthase 2 (NOS2) in vivo. Although NOS2 is easily induced by cytokines in primary cultured human airway epithelial cells and lung adenocarcinoma cell line A549, the human bronchial epithelial cell lines BEAS-2B and BET-1A do not express NOS2 in response to cytokines. Mechanisms regulating NOS2 expression in human respiratory epithelial cells are complex, but we have recently shown that NOS2 expression in primary human airway epithelial cells occurs in response to double-stranded RNA (dsRNA) through activation of signaling proteins including nuclear factor (NF) kappaB and interferon (IFN) regulatory factor (IRF)-1. In this context, we hypothesized that BEAS-2B and BET-1A cells may express NOS2 in response to dsRNA. Here, we show that although cytokines (IFN-gamma, tumor necrosis factor-alpha and interleukin-1beta) do not induce NOS2 expression in BEAS-2B or BET-1A cells, addition of dsRNA to this cytokine mix enables BEAS-2B cells to express NOS2. IFN gamma and dsRNA induction of NOS2 in BET-1A cells occurs in a serum concentration dependent manner, with a minimum of 3 d of serum treatment necessary for BET-1A cells to acquire the potential to induce NOS2. Importantly, dsRNA strongly activates NF-kappaB and IRF-1 in BEAS-2B cells, transcription factors essential for NOS2 gene expression in other cell lines. On the basis of these results, dsRNA-activated signaling pathways are clearly important for NOS2 expression in human respiratory epithelial cells. With conditions for NOS2 expression characterized, these cell lines are a convenient in vitro system to investigate the mechanisms regulating NOS2 expression in human respiratory epithelial cells. PMID- 11415938 TI - Prevention of polymerization of M and Z alpha1-Antitrypsin (alpha1-AT) with trimethylamine N-oxide. Implications for the treatment of alpha1-at deficiency. AB - alpha1-Antitrypsin (alpha1-AT) is the most abundant circulating proteinase inhibitor. The Z variant results in profound plasma deficiency as the mutant polymerizes within hepatocytes. The retained polymers are associated with cirrhosis, and the lack of circulating protein predisposes to early onset emphysema. We have investigated the role of the naturally occurring solute trimethylamine N-oxide (TMAO) in modulating the polymerization of normal M and disease-associated Z alpha1-AT. TMAO stabilized both M and Z alpha1-AT in an active conformation against heat-induced polymerization. Spectroscopic analysis demonstrated that this was due to inhibition of the conversion of the native state to a polymerogenic intermediate. However, TMAO did not aid the refolding of denatured alpha1-AT to a native conformation; instead, it enhanced polymerization. These data show that TMAO can be used to control the conformational transitions of folded alpha1-AT but that it is ineffective in promoting folding of the polypeptide chain within the secretory pathway. PMID- 11415939 TI - Building Elastin. Incorporation of recombinant human tropoelastin into extracellular matrices using nonelastogenic rat-1 fibroblasts as a source for lysyl oxidase. AB - The purpose of this study was to assess the feasibility of crosslinking exogenously produced tropoelastin, the precursor of insoluble elastin, into existing elastin. Tritiated recombinant human tropoelastin (rhTE) was added to neonatal rat aorta smooth-muscle cell (NNRSMC) cultures. As much as 12% of the added rhTE was incorporated into the NNRSMC-derived insoluble elastin with the formation of the elastin crosslinks desmosine (DES) and isodesmosine (IDES) in a time-dependent fashion. The ratio of radioactivity found in DES and IDES crosslinks to that found in lysyl residues increased from 0.18 immediately after incubation with rhTE to 0.76 after 14 d. Crosslinking of rhTE into elastin and the accompanying formation of tritiated water was inhibited by beta aminoproprionitrile, a potent inhibitor of lysyl oxidase, an enzyme critical for the post-translational processing of elastin and collagen. Acellular NNRSMC matrices were produced and replated with Rat-1 fibroblasts, cells that were found to express lysyl oxidase but not tropoelastin. At 14 d after incubation with rhTE, the ratio of DES and IDES radioactivity to that of lysine in the insoluble elastin was 0.38. We show for the first time that cells expressing lysyl oxidase, but not elastin, as well as elastogenic cells can incorporate rhTE into insoluble elastin with the formation of elastin crosslinks. This novel approach might be used to augment elastin repair in certain pathologic states. PMID- 11415940 TI - Quantitative trait analysis of nickel-induced acute lung injury in mice. AB - The genetic determinants underlying susceptibility to acute lung injury have not been identified. Recently, we found that the strain distribution pattern for mean survival time (MST) to three irritants-ozone, ultrafine Teflon, and nickel sulfate- was shared between inbred mouse strains. For ozone-induced acute lung injury, survival was found to be a complex trait controlled by at least three quantitative trait loci (QTLs), designated Aliq1, Aliq2, and Aliq3. To explore whether similar genes might be involved in survival to acute lung injury induced by nickel sulfate, we took advantage of the 2-fold difference in MSTs between the sensitive A/J and resistant C57BL/6J mice. QTL analysis of 307 backcross mice generated from these strains identified significant linkage to chromosome 6 (proposed as Aliq4) and suggestive linkage on chromosomes 1 and 12. Loci on chromosomes 9 and 16 had lod scores (log of the odds ratio, which equals the log of the "likelihood of linkage divided by the likelihood of no linkage") below significance, but contributed to the overall response. Comparing MSTs of backcross mice with similar haplotypes identified an allelic combination of four QTLs that could account for the survival time difference between the parental strains. Similar QTL intervals on chromosomes 6 and 12 were previously identified with ozone, suggesting that the interplay between different combinations of relatively few genes might be important for irritant-induced acute lung injury survival. PMID- 11415941 TI - Anti-proteinase 3 antibody activation of neutrophils can be inhibited by alpha1 antitrypsin. AB - Wegener's granulomatosis (WG) is classically associated with the presence of cytoplasmic antineutrophil cytoplasmic autoantibodies (c-ANCA). Proteinase 3 (PR3), the target antigen for c-ANCA, is inhibited by the antiprotease alpha1 antitrypsin (A1AT), and recent studies have demonstrated that WG patients who are A1AT-deficient have a worse clinical course, suggesting that a protease antiprotease imbalance may play a role in WG. We evaluated the effect of A1AT on anti-PR3 antibody-induced activation of neutrophils. The neutrophil was chosen because of its central role in the pathogenesis of WG. Isolated neutrophils from healthy controls were incubated with tumor necrosis factor (TNF)-alpha to induce surface expression of PR3. Subsequently, they were stimulated with a monoclonal antibody to PR3, resulting in a significant increase in respiratory burst. Addition of A1AT (1 mg/ml) to the TNF-alpha- primed cells before the addition of the anti-PR3 antibody resulted in a 47% reduction in anti-PR3 antibody-induced activation. A1AT mediated this inhibitory action by preventing anti-PR3 antibody binding to PR3 on the cell, thereby preventing the PR3-FcgammaR11a cross-linkage required for cell activation. Further, anti-PR3 antibody-induced activation of neutrophils from WG patients can be reduced by 56% with A1AT. These data suggest that protease-antiprotease interactions may play a pivotal role in neutrophil activation in WG. PMID- 11415942 TI - Interleukin-13 upregulates eotaxin expression in airway epithelial cells by a STAT6-dependent mechanism. AB - Interleukin (IL)-13 is a T helper 2-derived cytokine that has recently been implicated in allergic airway responses. We hypothesized that IL-13 may regulate expression of eotaxin in airway epithelium. We found that IL-13 upregulated eotaxin messenger RNA and protein synthesis in the airway epithelial cell line BEAS-2B; this effect showed synergy with tumor necrosis factor (TNF)-alpha and also was inhibited by the glucocorticoid budesonide. To establish the mechanisms of eotaxin upregulation by IL-13, cells were transfected with an eotaxin promoter luciferase reporter plasmid and transcription was activated by IL-13 (1.7-fold) and TNF-alpha (2.8-fold). The combination of IL-13 and TNF-alpha additively activated the promoter constructs (4.1-fold). Activation of signal transducer and activator of transcription (STAT) 6 by IL-13 was confirmed by nuclear protein binding to a DNA probe derived from the eotaxin promoter. Activation of eotaxin transcription by IL-13 and the additive effect with TNF-alpha were lost in plasmids mutated at a putative STAT6 binding site. Cotransfection with a wild type STAT6 expression vector significantly enhanced activation of the eotaxin promoter after IL-13 stimulation (6-fold induction). A significant increase of eotaxin protein secretion in the supernatant of STAT6 wild-type-transfected cells was observed after IL-13 stimulation. Cotransfection with a dominant negative STAT6 mutant expression vector inhibited activation of the eotaxin promoter by IL 13. These results indicate that IL-13 stimulates eotaxin expression in airway epithelial cells and that STAT6 plays a pivotal role in this response. PMID- 11415943 TI - Rapid reactive oxygen species production by mitochondria in endothelial cells exposed to tumor necrosis factor-alpha is mediated by ceramide. AB - Tumor necrosis factor (TNF)-alpha increases mitochondrial reactive oxygen species (ROS) production in tumor cells and hepatocytes. However, whether TNF-alpha stimulates mitochondrial ROS production in endothelial cells (EC) has not yet been reported. We studied the effect of TNF-alpha on mitochondrial ROS generation in EC and the signaling pathways involved. Cultured human umbilical vein EC (HUVEC) were studied by fluorescence microscopy, using dichlorodihydrofluorescein diacetate (DCFH-DA) as a marker of ROS production and propidium iodide uptake for cell viability. TNF-alpha increased DCFH oxidation in HUVEC dose-dependently. To determine the source of ROS, the mitochondrial respiratory chain inhibitors rotenone + thenoyltrifluoroacetone (TTFA), which inhibit electron entry to ubiquinone, and antimycin A (AA), a blocker of ubisemiquinone, were used. Rotenone and TTFA inhibited (n = 7, P < 0.05), whereas AA increased (118% in 3 min; n = 4, P < 0.01) ROS generation in HUVEC. In contrast, ROS production was not abolished by the nicotinamide adenine dinucleotide phosphate-dependent oxidase inhibitor diphenylene iodonium, by the xanthine oxidase inhibitor allopurinol, nor by the nitric oxide and cyclooxygenase pathway inhibitors N(omega)-nitro-L-arginine and mefenamic acid. In addition, TNF-alpha-induced ROS production was inhibited by the acidic sphingomyelinase inhibitor desipramine (5 microM; -80%, n = 4, P < 0.01) and totally blocked by the ceramide-activated protein kinase (CAPK) inhibitor dimethylaminopurine (1 mM; n = 6, P < 0.05). Thus, TNF-alpha induces mitochondrial ROS production in HUVEC that primarily occurs at the ubisemiquinone site and is mediated by ceramide-dependent signaling pathways involving CAPK. PMID- 11415944 TI - Hydrogen peroxide has opposing effects on IKK activity and IkappaBalpha breakdown in airway epithelial cells. AB - Recent studies have advanced our knowledge about the signal transduction cascade involved in the activation of nuclear factor (NF) kappaB, including the identification and characterization of IkappaB kinases (IKKs). Although exposure to hydrogen peroxide (H2O2) in vitro can activate NF-kappaB, this response is not universal and depends on the cell type and transformation state. In this study, we examined the effects of H2O2 on IKKs and activation of NF-kappaB in primary normal human bronchial epithelial (NHBE) cells. Our results demonstrate that treatment with H2O2 increased IKK activity, phosphorylation, and ubiquitination of IkappaBalpha in NHBE cells. However, there was no significant proteolytic degradation of IkappaBalpha, nuclear translocation of p65, or NF-kappaB DNA binding activity in cells treated with H2O2. Treatment with H2O2 also inhibited tumor necrosis factor (TNF)-alpha-induced IkappaBalpha breakdown, NF-kappaB DNA binding activity, and NF-kappaB-dependent transcription but had no effect on TNF alpha-induced IkappaBalpha phosphorylation or ubiquitination. Furthermore, treatment with H2O2 alone or in combination with TNF-alpha increased the levels of other ubiquitinated proteins in NHBE cells, suggesting general inhibition of proteasomal activity by H2O2. Taken together, these results demonstrate that in airway epithelial cells treatment with H2O2 has opposing effects on IKK activity and proteasomal degradation of IkappaBalpha, and suggest that H2O2 may suppress TNF-alpha-induced NF-kappaB- dependent gene expression. PMID- 11415945 TI - Molecular epidemiology: focus on infection. AB - Molecular biology techniques have become increasingly integrated into the practice of infectious disease epidemiology. The term "molecular epidemiology" routinely appears in the titles of articles that use molecular strain-typing ("fingerprinting") techniques-regardless of whether there is any epidemiologic application. What distinguishes molecular epidemiology is both the "molecular," the use of the techniques of molecular biology, and the "epidemiology," the study of the distribution and determinants of disease occurrence in human populations. The authors review various definitions of molecular epidemiology. They then comment on the range of molecular techniques available and present some examples of the benefits and challenges of applying these techniques to infectious agents and their affected host using tuberculosis and urinary tract infection as examples. They close with some thoughts about training future epidemiologists to best take advantage of the new opportunities that arise from integrating epidemiologic methods with modern molecular biology. PMID- 11415946 TI - Serum carotenoids and breast cancer. AB - The consumption of vegetables and fruit may protect against many types of cancer, but research evidence is not compelling for breast cancer. Carotenoids are pigments that are present in most plants and have known antioxidant properties. Blood concentrations of carotenoids have been proposed as integrated biochemical markers of vegetable, fruit, and synthetic supplements consumed. In a case control study (270 cases, 270 controls) nested within a cohort in New York during 1985-1994, the carotenoids lutein, zeaxanthin, beta-cryptoxanthin, lycopene, alpha-carotene, and beta-carotene were measured in archived serum samples using liquid chromatography. There was an evident increase in the risk of breast cancer for decreasing beta-carotene, lutein, alpha-carotene, and beta-cryptoxanthin. The risk of breast cancer approximately doubled among subjects with blood levels of beta-carotene at the lowest quartile, as compared with those at the highest quartile (odds ratio = 2.21; 95% confidence interval (CI): 1.29, 3.79). The risk associated with the other carotenoids was similar, varying between 2.08 (95% CI: 1.11, 3.90) for lutein and 1.68 (95% CI: 0.99, 2.86) for beta-cryptoxanthin. The odds ratio for the lower quartile of total carotenoids was 2.31 (95% CI: 1.35, 3.96). These observations offer evidence that a low intake of carotenoids, through poor diet and/or lack of vitamin supplementation, may be associated with increased risk of breast cancer and may have public health relevance for people with markedly low intakes. PMID- 11415947 TI - Invited commentary: serum carotenoids and breast cancer. PMID- 11415949 TI - Sexual factors and the risk of prostate cancer. AB - A population-based case-control study of prostate cancer was performed in King County, Washington, in White men and Black men aged 40-64 years, between 1993 and 1996. Incident prostate cancer cases (n = 753) were identified from the Seattle Puget Sound Surveillance, Epidemiology, and End Results (SEER) cancer registry. Controls (n = 703) were identified through random digit dialing and were frequency matched to cases on age. Sexual behavior, medical history, and other potential prostate cancer risk factors were ascertained through an in-person interview. There was no relation between sexual orientation and prostate cancer, although the number of men who had sex with men was small. Risk estimates increased directly with the lifetime number of female sexual partners (trend p < 0.001) but not with male partners (trend p = 0.62). Risk also increased with decreasing age at first intercourse, but this effect disappeared after adjusting for the number of female partners. Prior infection with gonorrhea was positively associated with risk (odds ratio = 1.50; 95% confidence interval: 1.0, 2.2), but no effect was seen among men with other sexually transmitted diseases. No relation between lifetime frequency of sexual intercourse and risk of prostate cancer was apparent. These findings are consistent with previous studies that support an infectious etiology for prostate cancer. PMID- 11415950 TI - Measures of menopausal status in relation to demographic, reproductive, and behavioral characteristics in a population-based study of women aged 35-49 years. AB - The purpose of this cross-sectional analysis of women aged 35-49 years from the Third National Health and Nutrition Examination Survey, conducted between 1988 and 1994, was to assess associations with menopausal status based either on menstrual cycle patterns or on elevated (>20 IU/liter) follicle-stimulating hormone. Menstrual cycle-based menopausal status was defined for women who had not had surgical menopause by months since the last period (<2, 2-12, and >12 months for pre-, peri-, and postmenopause, respectively). Logistic regression was adjusted for age, smoking, and unilateral oophorectomy. Higher body mass index (> or =30 kg/m(2) compared with < 25.0 kg/m(2)) was associated with a lower likelihood of elevated follicle-stimulating hormone (odds ratio (OR)=0.6, 95% confidence interval (CI): 0.4, 0.9) but this association was not seen with the menstrual measure of menopause. Exercise (three or more times per week) was associated with a lower likelihood of being postmenopausal on the basis of menstrual (OR = 0.3, 95% CI: 0.2, 0.7) and hormonal (OR = 0.6, 95% CI: 0.4, 1.0) measures. Alcohol use also tended to be associated with postmenopausal status by either measure, but not significantly so. There was little evidence of associations with ethnicity, education, age at menarche, number of livebirths, and oral contraceptive use. Menstrual-based definitions of menopause can be misclassified for women with menstrual irregularity. This might explain why obese women were classified menstrually as menopausal while remaining hormonally premenopausal. PMID- 11415951 TI - Influence of parity and lactation on hip fracture risk. AB - Several studies indicate that parity and lactation are associated with modest, short-term bone loss, but the long-term effect on osteoporotic fracture risk is uncertain. The authors therefore analyzed data from a population-based case control study among Swedish postmenopausal women aged 50-81 years between October 1993 and February 1995. Mailed questionnaires and telephone interviews were used to collect data on 1,328 incident cases with hip fracture and 3,312 randomly selected controls. In age-adjusted analyses, the risk of hip fracture among all women was reduced by 10% per child (95% confidence interval (CI): 5, 14). After multivariate adjustment including body mass index as a covariate, the risk reduction was 5% per child (95% CI: 0, 10). Oral contraceptive use modified the association of parity with hip fracture risk. Among never users of oral contraceptives, the risk of hip fracture was reduced by 8% per child (95% CI: 2, 13), whereas among ever users of oral contraceptives, the risk was in the opposite direction, with an increase in risk by 19% per child (95% CI: 0, 41). After parity was considered, there was no association of duration of lactation period with fracture risk. The authors conclude that parity is modestly associated with a reduced hip fracture risk among women who had not used oral contraceptives previously. PMID- 11415952 TI - Variability of angina symptoms and the risk of major ischemic heart disease events. AB - Variability of angina symptoms over a 5-year period was examined in a prospective study, in which 7,109 British middle-aged men completed two chest pain questionnaires, Q1 (1978-1980) and Q5 (1983-1985), and were classified as having no chest pain, nonexertional chest pain, or angina (Q) (exertional chest pain) on each occasion. Within persons, there was considerable variability in response to the chest pain questions at Q1 and Q5. Angina (Q) persistence showed marked associations with previous myocardial infarction, diagnosed angina, electrocardiogram ischemia, and subsequent major ischemic heart disease events from Q5 onward. Compared with men without angina (Q), the age-adjusted hazard ratios were 1.5 (95% confidence interval (CI): 1.1, 2.2) for angina (Q) at Q1 only, 2.6 (95% CI: 2.1, 3.2) for angina (Q) at Q5 only, and 3.4 (95% CI: 2.8, 4.3) for angina (Q) on both occasions. For men without diagnosed ischemic heart disease, for whom apparent remission of angina (Q) was particularly frequent, a similar pattern of association was found between angina (Q) persistence and subsequent major events. In middle-aged men, exertional chest pain is a strong indicator of major coronary risk but frequently appears transient in the longer term. Persistently reported symptoms are associated with severe disease and a poor prognosis. PMID- 11415953 TI - Distribution of serum C-reactive protein and its association with atherosclerotic risk factors in a Japanese population : Jichi Medical School Cohort Study. AB - The distribution of serum C-reactive protein (CRP) levels and their association with age, sex, and atherosclerotic risk factors were studied in a large Japanese population between 1992 and 1995. The subjects consisted of 2,275 males and 3,832 females aged 30 years and over. CRP was measured by nephelometry. The distribution of CRP was highly skewed toward a lower level than that of previous studies and seemed to be a combination of two separate distribution curves. The increase in CRP with age was statistically significant, and males had higher CRP levels than did females. Males who were current smokers had higher CRP levels than did nonsmokers. Age, systolic blood pressure, diastolic blood pressure, triglycerides, fibrinogen, and body mass index were all positively associated with CRP in both sexes, while total cholesterol and blood glucose were positively related in females only. High density lipoprotein cholesterol was inversely related in both sexes. Multiple logistic regression analysis showed that sex, age, systolic pressure, high density lipoprotein cholesterol, triglycerides, fibrinogen, and body mass index were significant independent variables. In conclusion, the distribution of CRP among the Japanese was quite different from that among Westerners, although CRP levels correlated with other atherosclerotic risk factors, similar to those in Westerners. PMID- 11415954 TI - Generational and regional differences in trends of mortality from ischemic heart disease in Japan from 1969 to 1992. AB - The authors compared generational and regional trends of premature mortality from ischemic heart disease (IHD) from 1969 to 1992 for persons aged 30-69 years. They selected Tokyo and Osaka prefectures as the most urbanized and compared them with the rest of Japan. The data were divided into two periods: period I (1969-1978, International Classification of Diseases, Eight Revision) and period II (1979 1992, International Classification of Diseases, Ninth Revision). In both populations, IHD mortality decreased for both sexes, but mortality from nonspecific heart disease remained constant in men and decreased in women. In Tokyo and Osaka prefectures, the percentage decline per year in IHD mortality for both sexes was significantly smaller in period II than in period I. However, in the rest of Japan, it did not decrease for either sex. Age-specific analysis showed that the percentage decline per year in period II was smallest for the group aged 30-49 years (men, 0.05%; women, 0.76%) in Tokyo and Osaka prefectures, while it was similar for all age groups in the rest of Japan. For men, the IHD mortality rate in 1991-1992 for those aged 30-49 years was higher in Tokyo and Osaka prefectures (9.4/100,000) than in the rest of Japan (5.4/100,000). PMID- 11415955 TI - Insulin resistance precedes weight loss in adults without diabetes : the Rancho Bernardo Study. AB - Insulin resistance is closely associated with both aging and overweight; yet in old age, weight loss is common, although insulin resistance increases. To study this paradox, the authors evaluated the role of insulin resistance in weight change among older adults from the Rancho Bernardo Study cohort. Participants were 725 nondiabetic men and women who were aged 50-89 years when weight and insulin were measured at baseline (1984-1987). The participants were evaluated again in 1992-1996, at which time weight was remeasured. Fasting insulin and homeostasis model assessment (HOMA) measurements were evaluated in separate but parallel statistical models as surrogates for insulin resistance. Insulin resistance, when defined as the top quartile of fasting insulin level or HOMA value, was significantly associated with weight loss before and after adjustment for baseline weight and age (fasting insulin: beta = -1.30 kg, p = 0.01; HOMA: beta = -1.18 kg, p = 0.01). Results were the same for men versus women, for the overweight (body mass index (weight (kg)/height (m)(2)) > 26.6) [corrected] versus the normal weight (body mass index < or = 26.6) [corrected], and for younger persons (age <70 years) versus older persons (age > or = 70 years). Insulin-resistant individuals had a threefold increased likelihood of losing 10 or more kg compared with those without insulin resistance. The authors conclude that hyperinsulinemia, independently of age and baseline weight, may have a catabolic effect in the elderly. PMID- 11415956 TI - Tofu consumption and blood lead levels in young Chinese adults. AB - Tofu is a commonly consumed food in China. Tofu may interfere with lead absorption and retention because of its high calcium content. In this observational study, the authors examined whether dietary tofu intake was associated with blood lead levels among young adults in Shenyang, China. The analyses included 605 men and 550 women who completed baseline questionnaires and had blood lead measurements taken in 1996-1998 as part of a prospective cohort study on reproductive health. Mean blood lead levels were 13.2 microg/dl in men and 10.1 microg/dl in women. Blood lead levels were negatively associated with tofu intake in both genders. A linear trend test showed a 3.7% (0.5-microg/dl) decrease in blood lead level with each higher category of tofu intake (p = 0.003). The highest tofu intake group (> or =750 g/week) had blood lead levels 11.3% lower (95% confidence interval: 4.1, 18.0) than those of the lowest tofu intake group (<250 g/week). In all regression models, data were adjusted for gender, age, height, body mass index, district, cigarette smoking, alcohol drinking, education, occupation, use of vitamin supplements, season, and dietary intake of meat, fish, vegetables, eggs, and milk. In conclusion, the authors found a significant inverse dose-response relation between tofu consumption and blood lead levels in this Chinese population. PMID- 11415957 TI - Use of generalized linear mixed models in the spatial analysis of small-area malaria incidence rates in Kwazulu Natal, South Africa. AB - Spatial statistical analysis of 1994-1995 small-area malaria incidence rates in the population of the northernmost districts of KwaZulu Natal, South Africa, was undertaken to identify factors that might explain very strong heterogeneity in the rates. In this paper, the authors describe a method of adjusting the regression analysis results for strong spatial correlation in the rates by using generalized linear mixed models and variograms. The results of the spatially adjusted, multiple regression analysis showed that malaria incidence was significantly positively associated with higher winter rainfall and a higher average maximum temperature and was significantly negatively associated with increasing distance from water bodies. The statistical model was used to produce a map of predicted malaria incidence in the area, taking into account local variation from the model prediction if this variation was supported by the data. The predictor variables showed that even small differences in climate can have very marked effects on the intensity of malaria transmission, even in areas subject to malaria control for many years. The results of this study have important implications for malaria control programs in the area. PMID- 11415958 TI - Commentary: practical advantages of Bayesian analysis of epidemiologic data. AB - In the past decade, there have been enormous advances in the use of Bayesian methodology for analysis of epidemiologic data, and there are now many practical advantages to the Bayesian approach. Bayesian models can easily accommodate unobserved variables such as an individual's true disease status in the presence of diagnostic error. The use of prior probability distributions represents a powerful mechanism for incorporating information from previous studies and for controlling confounding. Posterior probabilities can be used as easily interpretable alternatives to p values. Recent developments in Markov chain Monte Carlo methodology facilitate the implementation of Bayesian analyses of complex data sets containing missing observations and multidimensional outcomes. Tools are now available that allow epidemiologists to take advantage of this powerful approach to assessment of exposure-disease relations. PMID- 11415959 TI - Comparison of assessment methods for pesticide exposure in a case-control interview study. AB - In epidemiologic studies, much of the variation in disease risk estimates associated with occupational pesticide exposure may be due to variation in exposure classification. The authors compared five different methods of using interview information to assess occupational pesticide exposure in a US-Canada case-control study of neuroblastoma (1992-1994). For each method, exposure assignment was compared with that of a reference method, and neuroblastoma effect estimates were calculated. Compared with the reference method, which included a complete review of occupation, industry, job tasks, and exposure-specific activities, the use of occupation-industry groups alone or in combination with general job task information diluted the exposed group by including individuals who were unlikely to have been truly exposed. The effect estimates representing associations between each exposure method and neuroblastoma were different enough to influence the study's conclusions, especially when the exposure was rare (for maternal occupational pesticide exposure, the odds ratio was 0.7 using the reference exposure assessment method and 3.2 using the occupation-industry group exposure assessment method). Exposure-specific questions about work activities can help investigators distinguish truly exposed individuals from those who report exposure but are unlikely to have been exposed above background levels and from those who have not been exposed but are misclassified as exposed because of their employment in an occupation-industry group determined a priori to be exposed. PMID- 11415960 TI - Neuronal responses in area 7a to multiple-stimulus displays: I. neurons encode the location of the salient stimulus. AB - The primate posterior parietal cortex (PPC) plays an important role in representing and recalling spatial relationships and in the ability to orient visual attention. This is evidenced by the parietal activation observed in brain imaging experiments performed during visuo- spatial tasks, and by the contralateral neglect syndrome that often accompanies parietal lesions. Individual neurons in monkey parietal cortex respond vigorously to the appearance of single, behaviorally relevant stimuli, but little is known about how they respond to more complex visual displays. The current experiments addressed this issue by recording activity from single neurons in area 7a of the PPC in monkeys performing a spatial version of a match-to-sample task. The task required them to locate salient stimuli in multiple-stimulus displays and release a lever after a subsequent stimulus appeared at the same location. Neurons responded preferentially to the appearance of salient stimuli inside their receptive fields. The presence of multiple stimuli did not affect appreciably the spatial tuning of responses in the majority of neurons or the population code for the location of the salient stimulus. Responses to salient stimuli could be distinguished from background stimuli approximately 100 ms after the onset of the cue. These results suggest that area 7a neurons represent the location of the stimulus attracting the animal's attention and can provide the spatial information required for directing attention to a salient stimulus in a complex scene. PMID- 11415961 TI - Neuronal responses in area 7a to multiple stimulus displays: II. responses are suppressed at the cued location. AB - Everyday visual scenes contain a variety of stimuli that vary in their significance. The companion paper demonstrates that neurons in the posterior parietal cortex (PPC) are capable of encoding the spatial locations of the salient stimulus in multiple stimulus scenes. The present experiment sought to address how neuronal responses to stimuli appearing in the receptive field are modulated after attention has been drawn to one of multiple stimuli in a visual scene. We recorded from area 7a of the PPC in monkeys trained to do a spatial version of a match-to-sample task. The results show that neuronal responses are greatly suppressed when stimuli appear at previously attended locations. No reduction in responsiveness is observed for locations where stimuli had previously appeared but did not draw attention. These results support the hypothesis that area 7a has a role in redirecting attention to stimuli appearing at novel, unattended locations. PMID- 11415962 TI - A proton magnetic resonance spectroscopy study of age-related changes in frontal lobe metabolite concentrations. AB - Ageing is associated with reduction of grey matter volume and it is reported that the frontal lobes are preferentially affected. We have applied quantitative magnetic resonance spectroscopy (MRS), incorporating measurement of brain tissue water content and metabolite T(2) relaxation times, to determine absolute concentrations of the putative neuronal marker N-acetylaspartate (NAA), creatine (Cr) and choline (Cho) compounds in the frontal lobe of 50 male subjects aged between 20 and 70 years (10 per decade). The fractional brain water content (beta(MR)) did not change significantly as a function of age (r = 0.07, P = 0.65) and had a mean value of 81% (CV = 2%). The concentration (in millimoles per litre brain tissue) of NAA decreased significantly with age (r = -0.42, P = 0.003), with an overall decrease of 12% between the third and seventh decades. The concentrations of Cr and Cho did not change significantly with age. The interpretation of the age-dependent decrease in NAA concentration as reflecting either a reduction in neuronal volume, number or function is discussed. PMID- 11415963 TI - Parieto-frontal interactions in visual-object and visual-spatial working memory: evidence from transcranial magnetic stimulation. AB - This study aimed to investigate whether transcranial magnetic stimulation (TMS) can induce selective working memory (WM) deficits of visual-object versus visual spatial information in normal humans. Thirty-five healthy subjects performed two computerized visual n-back tasks, in which they were required to memorize spatial locations or abstract patterns. In a first series of experiments, unilateral or bilateral TMS was delivered on posterior parietal and middle temporal regions of both hemispheres after various delays during the WM task. Bilateral temporal TMS increased reaction times (RTs) in the visual-object, whereas bilateral parietal TMS selectively increased RTs in the visual-spatial WM task. These effects were evident at a delay of 300 ms. Response accuracy was not affected by bilateral or unilateral TMS of either cortical region. In a second group of experiments, bilateral TMS was applied over the superior frontal gyrus (SFG) or the dorsolateral prefrontal cortex (DLPFC). TMS of the SFG selectively increased RTs in the visual-spatial WM task, whereas TMS of the DLPFC interfered with both WM tasks, in terms of both accuracy and RTs. These effects were evident when TMS was applied after a delay of 600 ms, but not one of 300 ms. These findings confirm the segregation of WM buffers for object and spatial information in the posterior cortical regions. In the frontal cortex, the DLPFC appears to be necessary for WM computations regardless of the stimulus material. PMID- 11415964 TI - Odorants elicit evoked potentials in the human amygdala. AB - Electroencephalographical (EEG) recording studies have shown that odorants produce olfactory evoked potentials (OEPs) on the scalp surface. However, EEGs can only provide limited information about the intracerebral sources from where the OEPs are generated. By contrast, intracerebral EEG recordings enable direct examination of the electrophysiological activity from a given cerebral area. In the present study, neural activity was recorded from the amygdala of seven epileptic patients undergoing intracerebral EEG recordings prior to surgical treatment for relief of intractable seizures. Two olfactory tests were used: a passive-stimulation test consisting of the successive presentation of 12 common odorants and a suprathreshold detection test including both odorant and non odorant stimulations. Recordings from the amygdala revealed that all odorant stimulations induced large and reproducible OEPs, whereas the non-odorant stimulations did not. It was also found that repetition of the same odorant stimulation led to a decrease in the latency of the first OEP component. This modulation, which corresponds to a faster olfactory processing, strongly suggests that the amygdala is involved in early olfactory attentional processes. In conclusion, it appears that the human amygdala discriminates the incoming information from the nasal airflow as being odorant or not and, additionally, that its speed of processing is sensitive to recent experience with an odor. PMID- 11415965 TI - The role of the dorsolateral prefrontal cortex during sequence learning is specific for spatial information. AB - Many studies have implicated the dorsolateral prefrontal cortex in the acquisition of skill, including procedural sequence learning. However, the specific role it performs in sequence learning has remained uncertain. This type of skill has been intensively studied using the serial reaction time task. We used three versions of this task: a standard task where the position of the stimulus cued the response; a non-standard task where the color of the stimulus was related to the correct response; and a combined task where both the color and position simultaneously cued the response. We refer to each of these tasks based upon the cues available for guiding learning as position, color and combined tasks. The combined task usually shows an enhancement of skill acquisition, a result of being driven by two simultaneous and congruent cues. Prior to the performance of each of these tasks the function of the dorsolateral prefrontal cortex was disrupted using repetitive transcranial magnetic stimulation. This completely prevented learning within the position task, while sequence learning occurred to a similar extent in both the color and combined tasks. So, following prefrontal stimulation the expected learning enhancement in the combined task was lost, consistent with only a color cue being available to guide sequence learning in the combined task. Neither of these effects was observed following stimulation at the parietal cortex. Hence the critical role played by the dorsolateral prefrontal cortex in sequence learning is related exclusively to spatial cues. We suggest that the dorsolateral prefrontal cortex operates over the short term to retain and manipulate spatial information to allow cortical and subcortical structures to learn a predictable sequence of actions. Such functions may emerge from the broader role the dorsolateral prefrontal cortex has in spatial working memory. These results argue against the dorsolateral prefrontal cortex constituting part of the neuronal substrate responsible for general aspects of implicit or explicit sequence learning. PMID- 11415966 TI - Calculating direction maps from intrinsic signals revealed by optical imaging. AB - Previous optical imaging studies used the vector-summation (VS) method for calculating direction and orientation preference maps. However, for direction maps it often resulted in direction vectors which showed a steep angle to that of orientation vectors violating the 'aperture rule'. The present report provides a simple procedure for calculating direction preference maps using the 'electro- physiologist's ear' approach. This approach takes into account the strongest directional response component (vector-maximum, VM) in each pixel of the optical image, reminiscent of how electro- physiologists determine direction preference by audio-monitoring of the firing rate of neurons. The major advantage of this method is that the orthogonal relationship between orientation and direction preference vectors is preserved and that for most image pixels direction preference can be faithfully described by a single vector parameter. Here we used the VM method for calculating direction and the VS method for calculating orientation preference maps and quantified their spatial relationship. The results showed that, typically, an iso-orientation domain contained a pair of patches that preferred opposite directions orthogonal to the orientation. Rate-of change maps for direction revealed that virtually all direction discontinuity lines linked orientation centres. Close to orientation centres, direction discontinuity lines ran chiefly parallel with iso-orientation lines, whereas more remotely they had either parallel or perpendicular courses. PMID- 11415967 TI - Real and illusory contour processing in area V1 of the primate: a cortical balancing act. AB - It is known that neurons in area V2 (the second visual area) can signal the orientation of illusory contours in the primate. Whether area V1 (primary visual cortex) can signal illusory contour orientation is more controversial. While some electrophysiology studies have ruled out illusory signaling in V1, other reports suggest that V1 shows some illusory-specific response. Here, using optical imaging and single unit electrophysiology, we report that primate V1 does show an orientation-specific response to the 'abutting line grating' illusory contour. However, this response does not signal an illusory contour in the conventional sense. Rather, we find that illusory contour stimulation leads to an activation map that, after appropriate subtraction of real line signal, is inversely related to the real orientation map. The illusory contour orientation is thus negatively signaled or de-emphasized in V1. This 'activation reversal' is robust, is not due merely to presence of line ends, is not dependent on inducer orientation, and is not due to precise position of line end stimulation of V1 cells. These data suggest a resolution for previous apparently contradictory experimental findings. We propose that the de-emphasis of illusory contour orientation in V1 may be an important signal of contour identity and may, together with illusory signal from V2, provide a unique signature for illusory contour representation. PMID- 11415968 TI - Vertical bias in dendritic trees of non-pyramidal neocortical neurons expressing GAD67-GFP in vitro. AB - The neocortical neuropil has a strong vertical (orthogonal to pia) orientation, constraining the intracortical flow of information and forming the basis for the functional parcellation of the cortex into semi-independent vertical columns or 'modules'. Apical dendrites of excitatory pyramidal neurons are a major component of this vertical neuropil, but the extent to which inhibitory, GABAergic neurons conform to this structural and functional design is less well documented. We used a gene gun to transfect organotypic slice cultures of mouse and rat neocortex with the enhanced green fluorescent protein (eGFP) gene driven by the promoter for glutamic acid decarboxylase 67 (GAD67), an enzyme expressed exclusively in GABAergic cells. Many GAD67-GFP expressing cells were highly fluorescent, and their dendritic morphologies and axonal patterns, revealed in minute detail, were characteristic of GABAergic neurons. We traced 150 GFP-expressing neurons from confocal image stacks, and estimated the degree of vertical bias in their dendritic trees using a novel computational metric. Over 70% of the neurons in our sample had dendritic trees with a highly significant vertical bias. We conclude that GABAergic neurons make an important contribution to the vertical neocortical neuropil, and are likely to integrate synaptic inputs from axons terminating within their own module. PMID- 11415969 TI - Podium for the opposition? PMID- 11415970 TI - Comment on Peter Lachmann's article 'Stem cell research--why is it regarded as a threat?' in EMBO reports, March 2001. PMID- 11415971 TI - Is opposition to GM crops science or politics? An investigation into the arguments that GM crops pose a particular threat to the environment. PMID- 11415972 TI - Foot-and-mouth disease in Europe. FMD is economically the most important disease of farm animals. Its re-emergence in Europe is likely to have consequences that go beyond severe alterations of livestock production and trade. PMID- 11415973 TI - Blooming prospects? Humans have eaten seaweed for millennia; now microalgae are to be served up in a variety of novel health supplements, medicaments and preparations. PMID- 11415974 TI - The future of medicine. Centralised health and genetic databases promise to increase quality of health care while lowering costs. But to get there, many legal and social obstacles will have to be overcome to prevent abuse. PMID- 11415977 TI - Neuroscience from different angles. Student symposium: from genes to thoughts. PMID- 11415978 TI - A feast of membrane protein structures in Madrid. Workshop: Pumps, channels and transporters: structure and function. PMID- 11415979 TI - Disease-related versus polymorphic mutations in human mitochondrial tRNAs. Where is the difference? AB - A number of point mutations in human mitochondrial (mt) tRNA genes are correlated with a variety of neuromuscular and other severe disorders including encephalopathies, myopathies, cardiopathies and diabetes. The complexity of the genotype/phenotype relationships, the diversity of possible molecular impacts of the different mutations at the tRNA structure/function levels, and the exponential discovery of new mutations call for the search for unifying features. Here, the basic features (at the levels of primary and secondary structure) of 68 'pathogenic' mutations are compared with those of 64 'polymorphic' neutral mutations, revealing that these standard parameters for mutant analysis are not sufficient to predict the pathogenicity of mt tRNA mutations. Thus, case by case molecular investigation remains the only means of assessing the growing family of pathogenic mutations in mt tRNAs. New lines of research are suggested. PMID- 11415980 TI - Secured cutting: controlling separase at the metaphase to anaphase transition. AB - The final irreversible step in the duplication and distribution of genomes to daughter cells takes place at the metaphase to anaphase transition. At this point aligned sister chromatid pairs split and separate. During metaphase, cohesion between sister chromatids is maintained by the chromosomal multi-subunit cohesin complex. Here, I review recent findings as to how anaphase is initiated by proteolytic cleavage of the Scc1 subunit of cohesin. Scc1 is cleaved by a site specific protease that is conserved in all eukaryotes, and is now called 'separase'. As a result of this cleavage, the cohesin complex is destroyed, allowing the spindle to pull sister chromatids into opposite halves of the cell. Because of the final and irreversible nature of Scc1 cleavage, this reaction is tightly controlled. Several independent mechanisms seem to impose regulation on Scc1 cleavage, acting on both the activity of separase and the susceptibility of the substrate. PMID- 11415981 TI - An artificial transcription activator mimics the genome-wide properties of the yeast Pdr1 transcription factor. AB - We analysed the genome-wide regulatory properties of an artificial transcription activator in which the DNA-binding domain of the yeast transcription factor, Pdr1, was fused to the activation domain of Gal4 (Pdr1*GAD). This Pdr1*GAD chimera was put under the control of the inducible GAL1 promoter. DNA microarray analyses showed that all the target genes upregulated by the well-studied native gain-of-function Pdr1-3 mutant were similarly activated by the chimerical factor Pdr1*GAD upon galactose induction. Additionally, this kinetic approach led us not only to confirm previously published targets, but also to define a hierarchy among members of the Pdr1 regulon. Our observations prove, for the first time at the complete genome level, that the DNA-binding domain of Pdr1 is sufficient to guide its specificity. We propose that this approach could be useful for the study of new transcription factors identified in silico from sequenced organisms. Complete data are available at www.biologie.ens.fr/yeast-publi.html. PMID- 11415982 TI - Thyroid hormone regulates the obesity gene tub. AB - Thyroid hormone T3/T4 is a major regulator of energy metabolism in vertebrates, and defects in thyroid status are frequently associated with changes in body weight. It is demonstrated here that thyroid hormone regulates in vivo and in vitro the tub gene, which when mutated in tubby mice causes obesity, insulin resistance and sensory deficits. Hypothyroidism in rats altered tub mRNA and protein in discrete brain areas. These changes could be attributed to thyroid hormone deficiency since T3/T4 treatment restored normal tub expression. T3 also upregulated tub mRNA within 4-6 h in neuronal cells in culture, suggesting that T3 is a positive regulator of tub gene expression. Thus, these results establish a novel pathway of T3 action and provide an important molecular link between thyroid status and the tubby-associated syndrome. PMID- 11415983 TI - Phosphorylation disrupts the central helix in Op18/stathmin and suppresses binding to tubulin. AB - Protein phosphorylation represents a ubiquitous control mechanism in living cells. The structural prerequisites and consequences of this important post translational modification, however, are poorly understood. Oncoprotein 18/stathmin (Op18) is a globally disordered phosphoprotein that is involved in the regulation of the microtubule (MT) filament system. Here we document that phosphorylation of Ser63, which is located within a helix initiation site in Op18, disrupts the transiently formed amphipathic helix. The phosphoryl group reduces tubulin binding 10-fold and suppresses the MT polymerization inhibition activity of Op18's C-terminal domain. Op18 represents an example where phosphorylation occurs within a regular secondary structural element. Together, our findings have implications for the prediction of phosphorylation sites and give insights into the molecular behavior of a globally disordered protein. PMID- 11415984 TI - Two cyclophilin A homologs with shared and distinct functions important for growth and virulence of Cryptococcus neoformans. AB - Cyclophilin A is the target of the immunosuppressant cyclosporin A (CsA) and is encoded by a single unique gene conserved from yeast to humans. In the pathogenic fungus Cryptococcus neoformans, two homologous linked genes, CPA1 and CPA2, were found to encode two conserved cyclophilin A proteins. In contrast to Saccharomyces cerevisiae, in which cyclophilin A mutations confer CsA resistance but few other phenotypes, cyclophilin A mutations conferred dramatic phenotypes in C. neoformans. The Cpa1 and Cpa2 cyclophilin A proteins play a shared role in cell growth, mating, virulence and CsA toxicity. The Cpa1 and Cpa2 proteins also have divergent functions. cpa1 mutants are inviable at 39 degrees C and attenuated for virulence, whereas cpa2 mutants are viable at 39 degrees C and fully virulent. cpa1 cpa2 double mutants exhibited synthetic defects in growth and virulence. Cyclophilin A active site mutants restored growth of cpa1 cpa2 mutants at ambient but not at higher temperatures, suggesting that the prolyl isomerase activity of cyclophilin A has an in vivo function. PMID- 11415985 TI - Reconstitution of Sec-dependent membrane protein insertion: nascent FtsQ interacts with YidC in a SecYEG-dependent manner. AB - The inner membrane protein YidC is associated with the preprotein translocase of Escherichia coli and contacts transmembrane segments of nascent inner membrane proteins during membrane insertion. YidC was purified to homogeneity and co reconstituted with the SecYEG complex. YidC had no effect on the SecA/SecYEG mediated translocation of the secretory protein proOmpA; however, using a crosslinking approach, the transmembrane segment of nascent FtsQ was found to gain access to YidC via SecY. These data indicate the functional reconstitution of the initial stages of YidC-dependent membrane protein insertion via the SecYEG complex. PMID- 11415986 TI - Sec-dependent membrane protein insertion: sequential interaction of nascent FtsQ with SecY and YidC. AB - Recent studies identified YidC as a novel membrane factor that may play a key role in membrane insertion of inner membrane proteins (IMPs), both in conjunction with the Sec-translocase and as a separate entity. Here, we show that the type II IMP FtsQ requires both the translocase and, to a lesser extent, YidC in vivo. Using photo-crosslinking we demonstrate that the transmembrane (TM) domain of the nascent IMP FtsQ inserts into the membrane close to SecY and lipids, and moves to a combined YidC/lipid environment upon elongation. These data are consistent with a crucial role for YidC in the lateral transfer of TM domains from the Sec translocase into the lipid bilayer. PMID- 11415987 TI - A putative GDP-GTP exchange factor is required for development of the excretory cell in Caenorhabditis elegans. AB - The Caenorhabditis elegans excretory cell extends tubular processes, called canals, along the basolateral surface of the epidermis. Mutations in the exc-5 gene cause tubulocystic defects in this canal. Ultrastructural analysis suggests that exc-5 is required for the proper placement of cytoskeletal elements at the apical epithelial surface. exc-5 encodes a protein homologous to guanine nucleotide exchange factors and contains motif architecture similar to that of FGD1, which is responsible for faciogenital dysplasia. exc-5 interacts genetically with mig-2, which encodes Rho GTPase. These results suggest that EXC 5 controls the structural organization of the excretory canal by regulating Rho family GTPase activities. PMID- 11415988 TI - Angiostatin effects on endothelial cells mediated by ceramide and RhoA. AB - Angiostatin is a cleavage product of plasminogen that has anti-angiogenic properties. We investigated whether the effects of angiostatin on endothelial cells are mediated by ceramide, a lipid implicated in endothelial cell signaling. Our results demonstrate that angiostatin produces a transient increase in ceramide that correlates with actin stress fiber reorganization, detachment and death. DNA array expression analysis performed on ceramide-treated human endothelial cells demonstrated induction of certain genes involved in cytoskeleton organization. Specifically, we report that treatment with angiostatin or ceramide results in the activation of RhoA, an important effector of cytoskeletal structure. We also show that treatment of endothelial cells with the antioxidant N-acetylcysteine abrogates morphological changes and cytotoxic effects of treatment with angiostatin or ceramide. These findings support a model in which angiostatin induces a transient rise in ceramide, RhoA activation and free radical production. PMID- 11415989 TI - Minireview: transcriptional control of osteoblast differentiation. AB - The last 5 yr have witnessed major progress in skeleton biology. One of these areas of progress has been the partial elucidation of the transcriptional mechanisms of osteoblast differentiation and their conservation between mouse and human. Cbfal, or runx2, a homolog of the Drosophila Runt protein serves both as the earliest transcriptional regulator of osteoblast differentiation and as controller of bone formation by already differentiated osteoblast. Moreover, Cbfal is also a hypertrophic chondrocyte differentiation factor in several skeletal elements. Although other transcription factors are likely to be involved in osteoblast differentiation, Cbfal, by the multiplicity of the role that it plays, can be viewed as the central regulator of intramembranous and endochondral ossification. PMID- 11415990 TI - Vitamin D 1alpha-hydroxylase knockout mice as a hereditary rickets animal model. PMID- 11415991 TI - Differential effects of estradiol on the adrenocorticotropin responses to interleukin-6 and interleukin-1 in the monkey. AB - Endotoxin and the inflammatory cytokines interleukin (IL)-1 and IL-6 are potent activators of the hypothalamic-pituitary-adrenal (HPA) axis. Although estradiol (E(2)) has been shown to enhance the HPA response to certain types of stress, previous studies in the rodent have shown that HPA responses to endotoxin and to IL-1 were enhanced by ovariectomy and attenuated by E(2). The mechanisms underlying these observations are unclear, but there is evidence that E(2) may have direct inhibitory effects on IL-6 synthesis and release. Because endotoxin and IL-1 both stimulate IL-6, it is possible that the E(2)-induced suppression of the HPA response to endotoxin and IL-1 results from decreased IL-6 release. We have therefore examined the ACTH response to IL-6 and IL-1beta in six ovariectomized rhesus monkeys with and without 3 weeks of E(2) replacement. In the first study, plasma ACTH levels peaked at 60 min after iv injection of 6 microg recombinant human IL-6. Both the ACTH response, over time, and the area under the ACTH response curve were significantly higher in the E(2)-treated animals (P < 0.05). The peak ACTH level was 66 +/- 16 pg/ml without E(2) vs. 161 +/- 69 pg/ml with E(2). In the second study, iv infusion of recombinant human IL 1beta (400 ng) produced plasma IL-6 levels comparable with those seen after IL-6 injection in the first study. In the IL-1 study, however, there was a significant attenuation of the ACTH response, over time, in the E(2)-treated animals (P < 0.001); the peak ACTH level was 83 +/- 34 pg/ml vs. 13 +/- 4.4 pg/ml after E(2). The IL-6 response was similarly attenuated (P < 0.001); the peak IL-6 level was 614 +/- 168 pg/ml vs. 277 +/- 53 pg/ml after E(2) treatment. Our results demonstrate that physiological levels of E(2) enhance the ACTH response to IL-6 but attenuate the ACTH response to IL-1. The attenuated ACTH response to IL-1 was accompanied by a blunted IL-6 response. Our results suggest that the blunted HPA response to IL-1 can be explained, at least in part, by E(2)-induced alterations in IL-6 release. It remains to be determined whether E(2) affects other inflammatory mediators that also participate in this process. PMID- 11415992 TI - Analysis of ovarian gene expression in follicle-stimulating hormone beta knockout mice. AB - FSH is a heterodimeric glycoprotein hormone that is produced in the gonadotroph cells of the anterior pituitary. It acts on Sertoli cells of the testis and granulosa cells of the ovary. We previously demonstrated that FSHbeta knockout female mice are infertile due to a block in folliculogenesis preceding antral stage development. To investigate aberrations of ovarian gene regulation in the absence of FSH, we analyzed the expression of several important marker genes using Northern blot and in situ hybridization techniques. Key findings are as follows: 1) Follicles of FSHbeta knockout mice develop a well organized thecal layer, which is positive for P450 17alpha-hydroxylase and LH receptor messenger RNAs (mRNAs). This indicates that theca recruitment is completed autonomously with respect to FSH. 2) Granulosa cells in FSH-deficient mice demonstrate an increase in FSH receptor mRNA, and decreases in P450 aromatase, serum/glucocorticoid-induced kinase, and inhibin/activin subunit mRNAs. These data support studies that implicate FSH signaling cascades in the expression of these genes. 3) In contrast to the thecal layer, granulosa cell populations in FSHbeta knockout mice do not accumulate LH receptor mRNA. This suggests that although the granulosa cells have a block in proliferation at the antral follicle stage in the absence of FSH, they do not initiate programs of terminal differentiation as seen in luteinizing cells of wild-type ovaries. 4) Ovaries of FSH-deficient mice demonstrate a modest decrease in cyclin D2 mRNA, without up regulation of cell cycle inhibitor mRNAs associated with luteinization (i.e. p15, p27, and p21). Although components of the FSH null phenotype may be caused by partial cyclin D2 loss of function, these findings indicate that the mechanisms of granulosa cell cycle arrest in FSHbeta knockout mice are distinct from those of cycle withdrawal at luteinization. Underscoring the usefulness of the FSH deficient mouse model, this study clarifies aspects of gonadotropin-dependent folliculogenesis, thecal layer development, cycle control in granulosa cells, and luteinization. PMID- 11415993 TI - C-terminal parathyroid hormone-related protein (PTHrP) (107-139) stimulates intracellular Ca(2+) through a receptor different from the type 1 PTH/PTHrP receptor in osteoblastic osteosarcoma UMR 106 cells. AB - Studies were undertaken to determine whether PTH-related protein (PTHrP) (107 139) mobilizes [Ca(2+)](i) in osteoblastic osteosarcoma UMR 106 cells. PTHrP (107 139), in a manner similar to PTHrP (107-111), induced a rapid [Ca(2+)](i) response in these cells that was dose dependent (EC(50) of approximately 0.1 pM) and more efficient than that of PTHrP (1-36) (EC(50) of approximately 1 nM). This effect of PTHrP (107-139) was abrogated by micromolar doses of verapamil or nifedipine. However, it was unaffected by 10 microM U73122 (a phospholipase C inhibitor), 100 microg/ml heparin (an inositol 1,4,5-trisphosphate receptor inhibitor), or 400 ng/ml pertussis toxin (a G(i) inhibitor), which inhibited the [Ca(2+)](i) response to PTHrP (1-36), or by either 25 nM bisindolylmaleimide I (BIM), a protein kinase (PK) C inhibitor, or 1 microM phorbol-12-myristate-13 acetate preincubation (22 h). PTHrP (107-139) and PTHrP (1-36), at 100 nM, desensitized the [Ca(2+)](i) response to a second challenge with the same peptide, but not with the other peptide in these cells. PTHrP (7-34), a type 1 PTH/PTHrP receptor (PTH1R) antagonist, decreased the effect of PTHrP (1-36) on [Ca(2+)](i). In contrast, PTHrP (107-111), but neither PTHrP (109-138) nor PTHrP (7-34), abolished this effect of PTHrP (107-139). Both PTHrP (107-139) and PTHrP (1-36), added together at submaximal doses, induced a higher [Ca(2+)](i) response. Moreover, PTHrP (107-139) increased the efficacy of PTHrP (1-36) on [Ca(2+)](i), but decreased its induced increase in PKA activity in these cells. Verapamil or nifedipine (at 50 microM) or 25 nM BIM, but not 25 microM adenosine 3',5'-cyclic monophosphorothioate, Rp-isomer, a PKA inhibitor, abolished the PTHrP (107-139)-induced increase in interleukin 6 messenger RNA (assessed by RT, followed by PCR) in UMR 106 cells. This peptide also increased c-fos messenger RNA in these cells; an effect inhibited by BIM, but unaffected by either verapamil or EGTA. These findings support the existence of high-affinity receptors for PTHrP (107-139), associated with an induced Ca(2+) influx, different from the PTH1R in UMR 106 cells. The present results suggest that PTHrP could affect bone turnover by interacting with the PTH1R and other yet unknown receptors in bone cells through complex mechanisms. PMID- 11415994 TI - Effects of mutations involving the highly conserved S281HCC motif in the extracellular domain of the thyrotropin (TSH) receptor on TSH binding and constitutive activity. AB - A model has been proposed in which, in the absence of TSH, the extracellular domain of the TSH receptor would exert a silencing effect on the serpentine domain involved in activation of the G(alpha)(s) protein. Mutation of S281 in the ectodomain is supposed to release this constraint, thereby causing receptor activation. This defines S281 and its neighbors as a segment important in intramolecular signal transduction. The functional importance of this segment was explored by site-directed mutagenesis experiments involving S281, as well as the two cysteine residues (C283, C284) present immediately downstream. S281 was mutated to N, T, G, and A in this study, and the functional characteristics of the mutants were compared. We found that S281N, S281T, and S281G display stronger constitutive activity than S281A mutant, suggesting that increase in constitutive activity is related to the extent of disruption of the local structure of the ectodomain. C283 and C284, the two consecutive cysteines that are highly conserved in glycoprotein hormone receptors, were mutated to serine, either alone (S281HSC or S281HCS) or in combination (S281HSS) and were studied in two different TSH receptor backgrounds. The mutated cysteine ectodomains were either linked to a glycosylphosphatidylinositol anchor or the serpentine domain of the wild-type holoreceptor. Glycosylphosphatidylinositol-anchored ectodomain receptors showed good cell surface expression in CHO cells, but only S281HCS was able to bind TSH specifically, illustrating the importance of C283, or the putative disulphide bond, in maintaining the conformation of the ligand binding site. In contrast, cysteine mutants on an extracellular domain-holoreceptor background displayed severely impaired membrane targeting and were poorly expressed in COS cells. However, basal cAMP production, normalized to expression at the plasma membrane, indicated significant increase in constitutive activity of all three mutants, compared with the wild-type receptor. Altogether, these findings support a model in which the ectodomain would act as a silencer of the basal activity of the serpentine portion of the receptor. PMID- 11415995 TI - Leptin effects on immune function and energy balance are photoperiod dependent in Siberian hamsters (Phodopus sungorus). AB - Many adaptations have evolved in small mammals to maximize survival during winter. One such coping tactic in many species is an alteration of immune function in advance of the stressful conditions of winter. Leptin is a hormone produced by adipose tissue, and in addition to its central role in energy metabolism, leptin mediates the interactions among energy allocation, immune function, and reproduction. To examine this interaction further, exogenous leptin was administered for 2 weeks via osmotic minipumps to Siberian hamsters (Phodopus sungorus) housed in long or short days for a total of 12 weeks. Short-day hamsters displayed the expected reductions in humoral immune function, body mass, fat mass, and food intake. In Exp 1, exogenous leptin counteracted the reduction in food intake and the suppression of immune function in short days. In Exp 2, when the leptin-induced increase in food intake in short-day hamsters was prevented, leptin did not enhance immune function. In most of the measured fat pads and body mass, leptin had no effect in long days. In sum, leptin administered to short-day animals caused them to respond, in many cases, like long-day animals. Taken together, these data suggest that leptin acts indirectly to mediate energy allocation to humoral immune function. Additionally, leptin appears to act differentially, according to photoperiod, to regulate both immune and energetic parameters. PMID- 11415996 TI - Enhanced epidermal growth factor receptor signaling in MCF7 breast cancer cells after long-term culture in the presence of the pure antiestrogen ICI 182,780 (Faslodex). AB - This paper describes the establishment of an antiestrogen-resistant MCF7 breast cancer cell subline (FASMCF) by continuous culture of the estrogen-responsive parental line in steroid-depleted, ICI 182,780 (Faslodex; 10(-7) M)-supplemented medium. After a 3-month period of growth suppression, cells began to proliferate in ICI 182,780 at rates similar to those of untreated wild-type cells. Immunocytochemistry showed these cells to have reduced estrogen receptor and an absence of progesterone receptor proteins. RT-PCR and transient transfection studies with estrogen response element-reporter constructs confirmed that ICI 182,780-suppressed estrogen response element-mediated signaling. FASMCF cells show increased dependence upon epidermal growth factor receptor (EgfR)/mitogen activated protein kinase (MAPK)-mediated signaling. Thus, EgfR protein and messenger RNA, growth responses to transforming growth factor-alpha, and extracellular signal-regulated kinase 1/2 MAPK activation levels are all increased. Unlike wild-type cells, FASMCF cells are highly sensitive to growth inhibition by an EgfR-specific tyrosine-kinase inhibitor (TKI), ZD1839 (Iressa), and an inhibitor of the activation of MEK1 (MAPKK), PD098059. Short-term ( approximately 3 weeks) withdrawal of cells from antiestrogen had no effect on growth or phenotype, whereas longer withdrawal (>10 weeks) appeared to partially reverse the cellular phenotype with increasing estrogen receptor and decreasing EgfR levels. In subsequent studies FASMCF cells were maintained in TKI, where their growth was again suppressed and secondary TKI resistance failed to develop within the 3-month period in which initial ICI 182,780 resistance arose. Furthermore, wild-type cells similarly maintained in combination ICI 182,780 and TKI treatment conditions remained growth arrested (>6 months), with notable cell loss through both reduced rates of cellular proliferation and increased cell death. PMID- 11415997 TI - Testosterone inhibits spermatogonial differentiation in juvenile spermatogonial depletion mice. AB - The juvenile spermatogonial depletion (jsd) mutation results in spermatogonial arrest after the first wave of spermatogenesis. In homozygous jsd mice in a hybrid background (C3HxB6) that were identified with microsatellite markers, the percentage of tubules showing differentiating germ cells [tubule differentiation index (TDI)] rapidly decreased after 7 weeks of age with a correlative increase in the intratesticular testosterone (ITT) levels. Treatment with a GnRH antagonist, Cetrorelix, suppressed ITT and stimulated spermatogonial differentiation at the end of treatment. When treated mice were killed 5-13.3 weeks after the end of treatment, the ITT progressively increased, and the TDI progressively declined, but there was a transient appearance of tubules with mature spermatids. To delineate the role of testosterone (T) in spermatogonial arrest, we gave 7.6-week-old jsd mice exogenous T and/or the androgen receptor antagonist flutamide with or without GnRH antagonist for 4 weeks. Flutamide alone moderately stimulated spermatogonial differentiation (TDI = 30%). GnRH antagonist increased the TDI to 73%, and the addition of flutamide to the GnRH antagonist treatment further increased it to 95%. When T was combined with GnRH antagonist treatment, ITT was increased, and the TDI was reduced to 7%. Addition of flutamide to this combination reversed the T inhibition of GnRH antagonist stimulation of spermatogonial differentiation to a TDI of 57%. ITT levels showed a good negative correlation to the TDI obtained with various treatments, but no such correlation was observed for FSH or LH levels. The results indicate that T inhibits the ability of spermatogonia to differentiate in jsd mice through an androgen receptor-mediated process. PMID- 11415998 TI - Sucrose ingestion normalizes central expression of corticotropin-releasing-factor messenger ribonucleic acid and energy balance in adrenalectomized rats: a glucocorticoid-metabolic-brain axis? AB - Both CRF and norepinephrine (NE) inhibit food intake and stimulate ACTH secretion and sympathetic outflow. CRF also increases anxiety; NE increases attention and cortical arousal. Adrenalectomy (ADX) changes CRF and NE activity in brain, increases ACTH secretion and sympathetic outflow and reduces food intake and weight gain; all of these effects are corrected by administration of adrenal steroids. Unexpectedly, we recently found that ADX rats drinking sucrose, but not saccharin, also have normal caloric intake, metabolism, and ACTH. Here, we show that ADX (but not sham-ADX) rats prefer to consume significantly more sucrose than saccharin. Voluntary ingestion of sucrose restores CRF and dopamine-beta hydroxylase messenger RNA expression in brain, food intake, and caloric efficiency and fat deposition, circulating triglyceride, leptin, and insulin to normal. Our results suggest that the brains of ADX rats, cued by sucrose energy (but not by nonnutritive saccharin) maintain normal activity in systems that regulate neuroendocrine (hypothalamic-pituitary-adrenal), behavioral (feeding), and metabolic functions (fat deposition). We conclude that because sucrose ingestion, like glucocorticoid replacement, normalizes energetic and neuromodulatory effects of ADX, many of the actions of the steroids on the central nervous system under basal conditions may be indirect and mediated by signals that result from the metabolic effects of adrenal steroids. PMID- 11415999 TI - Prolactin induction of insulin gene transcription: roles of glucose and signal transducer and activator of transcription 5. AB - GH and PRL stimulate insulin production in pancreatic beta-cells through induction of insulin gene transcription. The transcriptional effects of GH are mediated through the binding of signal transducer and activator of transcription 5 (STAT5) to a consensus recognition sequence (TTCnnnGAA) in the rat insulin-1 promoter. In this study we demonstrate that PRL also induces the binding of STAT5 proteins to the rat insulin-1 STAT5 motif. However, the magnitude of binding of STAT5 nuclear proteins, as assessed by electrophoretic mobility shift assays, was only 1/30th that of the binding of the same STAT5 proteins to the beta-casein STAT5 site. The differences in the affinities of the rat insulin-1 and beta casein STAT5 motifs are explained in part by differences in promoter sequences flanking the STAT5 sites. To assess the importance of the STAT motif in PRL induction of insulin gene transcription, we deleted the STAT5 consensus sequence in the rat insulin 1 promoter, cloned the truncated promoter upstream of the luciferase reporter gene, and transfected the construct into rat insulinoma (INS 1) cells. The transcriptional activity of this construct was compared with that of the wild-type promoter. Although deletion of the STAT5 site in the promoter reduced the basal luciferase activity, the response to PRL was unaffected. PRL also induced transcription of constructs containing the wild-type human insulin promoter or the rat insulin-2 promoter, which contain no classic STAT5 sequences. The transcriptional effect of PRL was manifest even when cells were incubated in glucose-free medium, indicating that the action of the hormone is not mediated solely through changes in glucose uptake or glucose metabolism. To identify PRL responsive regions of the rat and human insulin promoters, we constructed a series of promoter truncations and assessed their responsiveness to PRL. A PRL responsive region of the rat insulin-1 promoter was localized between nucleotides -165 and -109. A PRL-responsive region of the human insulin promoter was localized between nucleotides -346 and -250. Additional regions of the human and rat insulin-1 promoters were required for PRL induction of a heterologous, minimal thymidine kinase promoter, suggesting that there are multiple PRL responsive elements in the insulin genes. These observations suggest a glucose- and STAT5-independent pathway by which PRL may induce insulin gene transcription. PMID- 11416000 TI - Involvement of mitogen-activated protein kinase in cyclic adenosine 3',5' monophosphate-induced hormone gene expression in rat pituitary GH(3) cells. AB - We examined whether mitogen-activated protein (MAP) kinase was activated by stimulation of the cAMP pathway and whether MAP kinase activation was involved in synthesis of PRL and GH in GH(3) cells. Treatment of the cells with a cAMP analog, 8-(4-chlorophenylthio)cAMP (CPT-cAMP), activated MAP kinase and increased PRL at both the protein and messenger RNA levels. The protein and messenger RNA of GH were decreased by the treatment. We constructed the luciferase reporter genes after the promoters of PRL and GH and found the activation of both promoters by the CPT-cAMP treatment. We confirmed that overexpression of the catalytic subunit of cAMP-dependent protein kinase had essentially the same effects on MAP kinase activation and synthesis of PRL and GH as the CPT-cAMP treatment. Furthermore, treatment of the cells with pituitary adenylate cyclase activating polypeptide 27 activated MAP kinase. The activation of PRL promoter by CPT-cAMP and pituitary adenylate cyclase-activating polypeptide 27 was abolished by pretreatment with PD098059 and H89. Although the increase in PRL and GH secretion by CPT-cAMP was inhibited by H89, PD098059 had no effect on secretion. These results suggest that cAMP-induced MAP kinase activation is essential for PRL gene expression, but not for secretion of PRL and GH. PMID- 11416001 TI - Identification of G protein-coupled, inward rectifier potassium channel gene products from the rat anterior pituitary gland. AB - Dopamine (DA) is a physiological regulator of PRL secretion, exerting tonic inhibitory control. DA activates an inward rectifier K(+) (IRK) channel in rat lactotropes, causing membrane hyperpolarization and inhibition of Ca(2+) dependent action potentials. Both the activation of this effector K(+) channel and the inhibition of PRL release are mediated by D(2)-type receptor activation and pertussis toxin- sensitive G proteins. To study the molecular basis of this physiologically relevant channel, a homology-based PCR approach was employed to identify members of the IRK channel family expressed in the anterior pituitary gland. Nondegenerate primers corresponding to regions specific for IRK channels known to be G protein activated (GIRKs; gene subfamily Kir 3.0) were synthesized and used in the PCR with reverse transcribed female rat anterior pituitary messenger RNA as the template. PCR products of predicted sizes for Kir 3.1, 3.2, and 3.4 were consistently observed by ethidium bromide staining after 16 amplification cycles. The identities of the products were confirmed by subcloning and sequencing. Expression of each of these gene products in anterior pituitary was confirmed by Northern blot analysis. Functional analysis of the GIRK proteins was performed in the heterologous expression system, Xenopus laevis oocytes. Macroscopic K(+) currents were examined in oocytes injected with different combinations of Kir 3.0 complementary RNA (cRNA) and G protein subunit (beta(1)gamma(2)) cRNA. The current-voltage relationships demonstrated strong inward rectification for each individual and pairwise combination of GIRK channel subunits. Oocytes coinjected with any pair of GIRK subunit cRNA exhibited significantly larger inward K(+) currents than oocytes injected with only one GIRK channel subtype. Ligand-dependent activation of only one of the GIRK combinations (GIRK1 and GIRK4) was observed when channel subunits were coexpressed with the D(2) receptor in Xenopus oocytes. Dose-response data fit to a Michaelis-Menten equation gave an apparent K(d) similar to that for DA binding in anterior pituitary tissue. GIRK1 and GIRK4 proteins were coimmunoprecipitated from anterior pituitary lysates, confirming the presence of native GIRK1/GIRK4 oligomers in this tissue. These data indicate that GIRK1 and GIRK4 are excellent candidate subunits for the D(2)-activated, G protein-gated channel in pituitary lactotropes, where they play a critical role in excitation-secretion coupling. PMID- 11416002 TI - Tyr(612) and Tyr(632) in human insulin receptor substrate-1 are important for full activation of insulin-stimulated phosphatidylinositol 3-kinase activity and translocation of GLUT4 in adipose cells. AB - To examine contributions of specific YXXM motifs in human insulin receptor substrate-1 (IRS-1) to mediating the metabolic actions of insulin, we studied IRS 1 mutants containing various substitutions of Phe for Tyr. In transfected NIH 3T3(IR) cells, insulin stimulation caused a 5-fold increase in phosphatidylinositol 3-kinase (PI3K) activity coimmunoprecipitated with wild-type IRS-1. No PI3K activity was associated with IRS1-F6 (Phe substituted for Tyr at positions 465, 612, 632, 662, 941, and 989). Adding back both Tyr(612) and Tyr(632) fully restored IRS-1-associated PI3K activity, whereas adding back either Tyr(612) or Tyr(632) alone was associated with intermediate PI3K activity. In rat adipose cells transfected with epitope-tagged GLUT4, insulin stimulation caused a 2-fold increase in cell surface GLUT4-HA. Cotransfection of cells with GLUT4-HA and either wild-type IRS-1 or IRS1-Y612/Y632 increased basal cell surface GLUT4-HA (in the absence of insulin) to approximately 80% of the levels seen in insulin-stimulated control cells, whereas overexpression of IRS1-F6 had no effect on the insulin dose-response curve. Overexpression of IRS1-Y612 or IRS1 Y632 caused intermediate effects. Thus, both Tyr(612) and Tyr(632) are important for IRS-1 to fully activate PI3K and mediate translocation of GLUT4 in response to insulin. PMID- 11416003 TI - The maternal diet during pregnancy programs altered expression of the glucocorticoid receptor and type 2 11beta-hydroxysteroid dehydrogenase: potential molecular mechanisms underlying the programming of hypertension in utero. AB - Potential mechanisms underlying prenatal programming of hypertension in adult life were investigated using a rat model in which maternal protein intake was restricted to 9% vs. 18% casein (control) during pregnancy. Maternal low protein (MLP) offspring exhibit glucocorticoid-dependent raised systolic blood pressure throughout life (20-30 mm Hg above the control). To determine the molecular mechanisms underlying the role of alterations in glucocorticoid hormone action in the prenatal programming of hypertension in MLP offspring, tissues were analyzed for expression of the glucocorticoid receptor (GR), mineralocorticoid receptor (MR), 11betaHSD1, 11betaHSD2, and corticosteroid-responsive Na/K-adenosine triphosphatase alpha1 and beta1. GR protein (95 kDa) and messenger RNA (mRNA) expression in kidney, liver, lung, and brain was more than 2-fold greater in MLP vs. control offspring during fetal and neonatal life and was more than 3-fold higher during subsequent juvenile and adult life (P < 0.01). This was associated with increased levels of Na/K-adenosine triphosphatase alpha1- and beta1-subunit mRNA expression. Levels of MR gene expression remained unchanged. Exposure to the MLP diet also resulted in markedly reduced levels of 11betaHSD2 expression in the MLP placenta on days 14 and 20 of gestation (P < 0.001), underpinning similar effects on 11betaHSD2 enzyme activity that we reported previously. Levels were also markedly reduced in the kidney and adrenal of MLP offspring during fetal and postnatal life (P < 0.001). This programmed decline in 11betaHSD2 probably contributes to marked increases in glucocorticoid hormone action in these tissues and potentiates both GR- and MR-mediated induction of raised blood pressure. In contrast, levels of 11betaHSD1 mRNA expression in offspring central and peripheral tissues remained unchanged. In conclusion, we have demonstrated that mild protein restriction during pregnancy programs tissue-specific increases in glucocorticoid hormone action that are mediated by persistently elevated expression of GR and decreased expression of 11betaHSD2 during adult life. As glucocorticoids are potent regulators not only of fetal growth but also of blood pressure, our data suggest important potential molecular mechanisms contributing to the prenatal programming of hypertension by maternal undernutrition in the rat. PMID- 11416004 TI - Maternal undernutrition during early to midgestation programs tissue-specific alterations in the expression of the glucocorticoid receptor, 11beta hydroxysteroid dehydrogenase isoforms, and type 1 angiotensin ii receptor in neonatal sheep. AB - We have investigated the effects of maternal nutrient restriction in the sheep during the period of rapid placental growth (i.e. 28-77 days gestation; term = 147 days) on feto-placental growth and expression of the glucocorticoid receptor (GR), types 1 and 2 11beta-hydroxysteroid dehydrogenase (11betaHSD1, 11betaHSD2), and types 1 and 2 angiotensin II receptor (AT1, AT2) in fetal and neonatal offspring. Ewes (n = 63) of similar age, body weight, and body composition were randomly allocated to a nutrient-restricted (NR) group in which they consumed 3.2 MJ/day metabolizable energy (ME; equivalent to 50% of predicted requirements) or to a control group in which they consumed 6.7 MJ/day ME (equivalent to 110% of predicted requirements). After 77 days gestation, ewes from both dietary groups consumed close to 100% of ME requirements up to term. Newborn offspring of NR ewes were of similar body weight, but had increased crown-rump length, greater placental weight, and increased placental/body weight ratio (P < 0.01) compared with controls. Their kidneys were heavier (P < 0.05), but shorter in length, with increased ratios of transverse width to length (P < 0.001). GR messenger RNA (mRNA) expression in neonatal offspring from NR ewes was increased in adrenal, kidney, liver, lung, and perirenal adipose tissue (P < 0.01). Conversely, 11betaHSD1 mRNA expression was unaffected, except in perirenal adipose tissue, where it was higher in lambs born to NR ewes (P < 0.01). 11betaHSD2 mRNA expression was decreased in adrenals and kidney (P < 0.001). Maternal NR also resulted in significantly increased AT1 expression in those tissues in which expression of GR was increased and/or 11betaHSD2 was decreased, i.e. adrenals, kidney, liver, and lung. AT2 expression was unaffected by maternal NR. Although 11betaHSD2 mRNA was undetectable in term placenta, it was abundant in midgestation placenta and was lower after maternal NR (P < 0.001). There was close agreement between levels of 11betaHSD enzyme (i.e. 11beta-dehydrogenase and 11-oxoreductase) activities and abundance of 11betaHSD1 mRNA and 11betaHSD2 mRNA expression. The persistence of tissue-specific increases in the expression of GR, 11betaHSD1 and AT1 and decreases in the expression of 11betaHSD2 in adrenals and kidney in newborn offspring in response to a defined period of maternal nutrient restriction during early to midgestation suggests that gene expression has been programmed by nutrient availability to the fetus before birth. These data suggest key potential mechanisms by which maternal nutrition prenatally programs physiological pathways, such as the renin-angiotensin system, in the offspring that may lead to raised blood pressure and other cardiovascular disease risk factors in later life. PMID- 11416005 TI - L-type Ca(2+) channel regulation by pituitary adenylate cyclase-activating polypeptide in vascular myocytes from spontaneously hypertensive rats. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP), a vasoactive peptide, modulates the L-type Ca(2+) channel current (L channel current) in vascular smooth muscle cells (VSMC) through activation and integration of two intracellular pathways, protein kinase A and protein kinase C (PKC). In the present study we compared the effects of PACAP on the L channel current in VSMC from the spontaneously hypertensive rats (SHR) and normotensive controls, Wistar Kyoto rats (WKY). We found that compared with WKY, VSMC from SHR had a higher L channel current density. Stimulation by PACAP (10 nM) caused an increase in the amplitude of the whole cell current and prolonged open time in VSMC from SHR and WKY, with the increase greater in SHR. These effects of PACAP on the L channel current was mimicked by an activator of PKC. In contrast, PACAP caused a smaller increase in cAMP accumulation in VSMC from SHR than WKY, and there was no difference in the inhibitory effect of 8-bromo-cAMP on the L channel current from both type of cells. The greater increase in amplitude of the L channel current by PACAP in VSMC from SHR persisted in the presence of adenosine cyclic 3',5' monophosphothioate, Rp-isomer, a cAMP antagonist, but not calphostin C, a PKC inhibitor. Taken together, our results show an increase in L channel current density and an enhanced PACAP effect on the L channel current in VSMC from SHR compared with WKY. This difference in PACAP response appears to be predominately secondary to an increased PKC sensitivity. PMID- 11416006 TI - Pituitary follistatin and activin gene expression, and the testicular regulation of FSH in the adult Rhesus monkey (Macaca mulatta). AB - In rats, FSHbeta gene expression and FSH secretion are increased and decreased, respectively, by pituitary activin and follistatin. Because little information is available on the paracrine control of FSH secretion in the primate, follistatin and activin/inhibin beta(B) messenger RNA (mRNA) levels were measured in pituitaries of adult male rhesus monkeys 6 weeks after castration or sham surgery (n = 5/group). Follistatin mRNA was determined by quantitative RT-PCR assay using oligonucleotide primers designed to span exons 3-5 of the human follistatin gene. Activin/inhibin beta(B) mRNA levels were measured by ribonuclease protection. Orchidectomy resulted in a 100-fold increase in plasma FSH concentrations and a 60-fold rise in those of LH. In castrated monkeys, levels of mRNA encoding FSHbeta, LHbeta, alpha- subunit, and GnRH receptor (GnRH-R) were increased 21-, 2.1-, 1.7-, and 1.7-fold, respectively (P < 0.01). Levels of pituitary follistatin and activin/inhibin beta(B) mRNAs, however, were similar in castrated and intact animals. These data suggest that the paracrine control of FSH secretion in the male differs substantially in primates and rodents. Specifically, the relatively greater postcastration rise in FSHbeta gene expression and FSH secretion in the adult male monkey may result because in this species pituitary follistatin gene expression does not increase after orchidectomy, as it does in the rat. PMID- 11416007 TI - Cloning of the ovine estrogen receptor-alpha promoter and functional regulation by ovine interferon-tau. AB - Interferon-tau (IFNtau), the ruminant pregnancy recognition signal, inhibits transcription of the estrogen receptor alpha (ERalpha) gene in the endometrial lumenal epithelium of the sheep uterus, thereby abrogating production of luteolytic PGF(2alpha) pulses. The effects of IFNtau are mediated in part by IFN stimulated response elements (ISREs) and IFN regulatory factor elements (IRFEs). The promoter/enhancer region of the ovine ERalpha gene was cloned, sequenced, and predicted to contain four IRFEs and one ISRE. Electrophoretic mobility shift assays indicated that the -2110 IRFE bound only IRF-1, whereas the -1877 IRFE and the -1284 ISRE were functional in binding IRF-1 and IRF-2. IFNtau inhibited transcriptional activity of the 2.7-kb ovine ERalpha promoter in transfection assays using ovine lumenal epithelium cells. Analyses of sequential 5'-deletion mutants of the ovine ERalpha promoter indicated that the effects of IFNtau may be mediated by IRFEs as well as other elements. Overexpression of ovine IRF-2, but not IRF-1, inhibited transcriptional activity of several regions of the ovine ERalpha promoter containing an IRFE or an ISRE as well as some, but not all, regions lacking these elements. PMID- 11416008 TI - Leptin administration prevents spontaneous gestational diabetes in heterozygous Lepr(db/+) mice: effects on placental leptin and fetal growth. AB - Gestational diabetes mellitus (GDM) results from an interaction between susceptibility genes and the diabetogenic effects of pregnancy. During pregnancy, mice heterozygous for the lepin receptor (db/+) gain more weight, are glucose intolerant, and produce macrosomic fetuses compared with wild-type (+/+) mothers, suggesting that an alteration in leptin action may play a role in GDM and fetal overgrowth. To investigate whether leptin administration or pair-feeding can reduce adiposity and thereby prevent GDM and neonatal overgrowth, we examined energy balance, glucose and insulin tolerance, and fetal growth in pregnant db/+ and +/+ mice treated with recombinant human leptin-IgG during late pregnancy. Leptin reduced food intake and adiposity in pregnant db/+ mice to levels similar to pregnant +/+ mice and significantly reduced maternal weight gain. Maternal glucose levels were markedly lower during glucose and insulin challenge tests in leptin-treated db/+ mice relative to db/+ and pair-fed controls. Despite reduced energy intake and improved glucose tolerance, leptin administration did not reduce fetal overgrowth in offspring from db/+ mothers. Fetal and placental leptin levels were 1.3- to 1.5-fold higher in offspring from db/+ mothers and remained unchanged with leptin administration, whereas leptin treatment in +/+ mothers or pair-feeding decreased placental leptin concentration and reduced fetal birth weight. Our results provide evidence that leptin administration during late gestation can reduce adiposity and improve glucose tolerance in the db/+ mouse model of spontaneous GDM. However, fetal and placenta leptin levels are higher in db/+ mothers and are subject to reduced negative feedback in response to leptin treatment. These data suggest that alterations in placenta leptin may contribute to the regulation of fetal growth independently of maternal glucose levels. PMID- 11416009 TI - Osteoclasts formed by measles virus-infected osteoclast precursors from hCD46 transgenic mice express characteristics of pagetic osteoclasts. AB - Pagetic osteoclasts (OCLs) are abnormal in size and contain paramyxoviral-like nuclear inclusions that cross-react with antibodies to measles virus (MV). However, the role that MV infection plays in Paget's disease is unknown, because no animal model of Paget's disease is available. Therefore, we targeted a cellular MV receptor, human CD46 (hCD46), to cells in the OCL lineage in transgenic mice using the mouse tartrate-resistant acid phosphatase (TRAP) gene promoter. In vitro infection of OCL precursors from hCD46 transgenic mice with MV significantly increased OCL formation in bone marrow cultures. The numbers of TRAP-positive mononuclear cells and CFU-GM, the earliest identifiable OCL precursor, were also significantly increased. MV-infected OCLs formed from hCD46 marrow were increased in size, contained markedly increased numbers of nuclei, and had increased bone-resorbing capacity per OCL compared with OCLs formed from marrow of nontransgenic littermates. Furthermore, IL-6 and 24-hydroxylase messenger RNA expression levels were increased in MV-infected hCD46 transgenic mouse bone marrow cultures. Treatment of MV-infected hCD46 marrow cultures with a neutralizing antibody to IL-6 blocked the increased OCL formation seen in these cultures. These data demonstrate that MV infection of OCL precursors results in OCLs that have many features of pagetic OCLs, that the enhanced OCL formation is in part mediated by increased IL-6 expression induced by MV infection, and suggest that the hCD46 transgenic mouse may be a useful model for examining the effects of MV infection on OCL formation in vivo. PMID- 11416010 TI - Polymorphic forms of expressed bovine interferon-tau genes: relative transcript abundance during early placental development, promoter sequences of genes and biological activity of protein products. AB - Multiple interferon (IFN)-tau genes exist in cattle, but it has remained unclear how many are expressed, the extent of their variation, and whether different genes exhibit similar patterns of expression and code for proteins with similar biological activities. A total of 118 complementary DNA (cDNA) were bi directionally sequenced from reverse-transcribed bovine (bo) conceptus RNA over the period from blastocyst formation until day 25 of pregnancy. Fourteen different cDNAs, encoding eight different IFN-tau, were confirmed unique. All showed high sequence conservation (>98% nucleotide identity; >96% amino acid identity). The cDNA fell into three, recently evolved, phylogenetic groups (tau1, 2, and 3). Mean concentrations of IFN-tau messenger RNA were greater at day 17 and day 19 than at day 14 and day 25, with different genes showing comparable expression patterns, although there appeared to be a major bias in expression of two genes (for boIFN-tau1c and tau3a) in blastocysts. Genes representing members of the three boIFN-tau groups were cloned. Their promoter regions were conserved over regions considered important for transcriptional activation. Recombinant protein generated in Escherichia coli from representative genes in the three groups had similar but not identical antiviral activities. In summary, many IFN tau genes, which are probably under similar transcriptional control, are expressed in bovine trophoblast during the peri-implantation period of development. PMID- 11416011 TI - Analysis of the testicular phenotype of the follicle-stimulating hormone beta subunit knockout and the activin type II receptor knockout mice by stereological analysis. AB - This study evaluated the role of FSH and activin A on testicular function using quantitative stereological analysis of testicular cell types in mice with targeted disruption of genes encoding the FSH beta-subunit and the activin type IIA receptor (ActRIIA). Using the optical dissector technique, the numbers of Sertoli cells and germ cells per testis were determined. Testis weights in homozygous males lacking the FSHbeta gene or the ActRIIA gene were decreased approximately 60% compared with wild-type or respective heterozygotes. Sertoli cell numbers decreased in both homozygous mice by 30-39%, and there was a comparable decline in germ cell numbers in both models. The degree of germ cell attrition increased in the later stages of spermatogenesis from a 46% reduction of spermatogonia to a 60% decrease in round spermatids. As the FSH levels are decreased in both models, the cellular lesion in both is most likely due to the FSH deficiency. Although the decrease in the Sertoli cell complement represents one cause of lower germ cell numbers, the ability of Sertoli cells to nurture germ cells is compromised by the lower FSH levels, as shown by a decrease in the round spermatid to Sertoli cell ratios in both homozygous models. We conclude that the defects in FSH beta-subunit gene knockout and ActRIIA knockout mice are related to diminished FSH action on both Sertoli cell proliferation and the capacity of Sertoli cells to nurture germ cells. PMID- 11416012 TI - Concerted transcriptional activation of the low density lipoprotein receptor gene by insulin and luteinizing hormone in cultured porcine granulosa-luteal cells: possible convergence of protein kinase a, phosphatidylinositol 3-kinase, and mitogen-activated protein kinase signaling pathways. AB - Insulin and insulin-like growth factor I (IGF-I) can amplify gonadotropin stimulated steroidogenesis by augmenting the expression of key sterol regulatory genes in ovarian cells, viz. low density lipoprotein (LDL) receptor, steroidogenic acute regulatory protein, and P450 cholesterol side-chain cleavage enzyme (CYP11A). The mechanisms underlying the foregoing bihormonal interactions are not known. Accordingly, in relation to the LDL receptor gene, the present study tests the hypothesis that insulin/IGF-I and LH can act via concerted transcriptional control of promoter expression. To this end, we transiently transfected primary monolayer cultures of porcine granulosa-luteal cells with a reporter vector containing the putative 5'-upstream full-length (pLDLR1076/luc) regulatory region (-1076 to +11 bp) of the homologous LDL receptor gene driving firefly luciferase in the presence or absence of insulin (or IGF-I) and/or LH (each 100 ng/ml). Combined exposure to LH and insulin (or IGF-I) stimulated LDL receptor transcriptional activity maximally at 4 h by 8- to 20-fold, as normalized by coexpression of Renilla luciferase. Further analysis of multiple 5' nested deletional constructs of the LDL receptor gene promoter showed that deletion of -139 bp upstream of the transcriptional start site virtually abolished basal expression and promoter responsiveness to LH and insulin/IGF-I. In contrast, full basal activity and 60-80% of maximal monohormonal and bihormonal drive were retained by the -255 to +11 bp fragment. As LDL receptor gene expression in other tissues is negatively regulated by the abundance of intracellular free cholesterol, we assessed the impact of concomitant pretreatment of granulosa-luteal cells with an exogenous soluble sterol (25 hydroxycholesterol, 1 and 10 microM). Excess sterol markedly (50-70%) attenuated bihormonally and, in lesser measure, LH-stimulated and basal LDL receptor promoter expression, thus affirming a feedback-sensitive sterol-repressive region in this gene. Non-LH receptor-dependent agonists of protein kinase A (PKA), 8 bromo-cAMP (1 mM), and forskolin (10 microM) with or without insulin/IGF-I costimulation likewise augmented LDL receptor promoter expression with similar strong dependency on the -255 to -139 bp 5'-upstream region. To assess more specific PKA-dependent mediation of LH's contribution to combined hormonal drive, the LDL receptor (-1076 to +11 bp) reporter plasmid was cotransfected with a full sequence rabbit muscle protein kinase inhibitor (PKI) minigene driven constitutively by a Rous sarcoma virus promoter. Expression of the latter PKA antagonist blocked transcriptional stimulation by LH alone as well as that by LH combined with insulin (or IGF-I) by 70-85% without reducing basal transcriptional activity. Transfection of a mutant inactive (Arg to Gly) Rous sarcoma virus/PKI gene confirmed the specificity of the PKI effect. To investigate the convergent role of the insulin/IGF-I effector pathway mediating bihormonal stimulation of LDL receptor promoter expression, transfected granulosa-luteal cells were pretreated for 30 min with two specific inhibitors of phophatidylinositol 3 kinase, wortmannin (100 nM) and LY 294002 (10 microM), or of mitogen-activated protein kinase kinase, PD 98059 (50 microM), U0126 (10 microM), or the latter's inactive derivative, U0124 (10 microM). Both classes of antagonists impeded the ability of insulin or IGF-I to enhance LH-stimulated LDL receptor promoter expression by 60-80%. In conclusion, the present analyses indicate that LH and insulin (or IGF-I) can up-regulate LDL receptor transcriptional activity supraadditively in porcine granulosa-luteal cells 1) via one or more agonistic cis-acting DNA regions located between -255 and -139 bp 5'- upstream of the transcriptional start site, 2) without abrogating sterol-sensitive repressive of this promoter, and 3) by way of intracellular mechanisms that include the PKA, phophatidylinositol 3-kinase, and mitogen-activated protein kinase signaling pathways. PMID- 11416013 TI - In vivo gonadotropin-releasing hormone secretion in female rats during peripubertal development and on proestrus. AB - Pubertal development in female rats is characterized by increased LH levels and the appearance of estrogen-dependent afternoon LH mini-surges. In these studies we performed the first analysis of GnRH patterns in peripubertal rats to determine whether there are similar changes in pulsatile GnRH release. Microdialysis samples were collected at 5-min intervals throughout a 5-h afternoon period from 22 rats sampled on a single day between 30-47 days of age. Adult female rats were sampled on proestrus for comparison. In 30- to 33-day-old rats, GnRH release was infrequent (2.7 pulses/5 h; n = 3), whereas intermediate pulse frequencies were observed in 34- to 37-day-old rats (6.4 pulses/5 h; n = 9) and 38- to 42-day-old (5.0 pulses/5 h; n = 5) rats. The highest GnRH pulse frequencies were observed in 43- to 47-day-old rats (9.4 pulses/5 h; n = 5). Mean GnRH pulse amplitude did not vary significantly with age. Animals sampled before vaginal opening (VO) exhibited significantly slower GnRH pulse frequencies than those sampled after vaginal opening (1.3 pulses/5 h pre-VO vs. 7.6 pulses/5 h post-VO; P = 0.01). An afternoon increase in GnRH secretion, defined operationally as a greater than 25% increase in mean GnRH levels in the last half of the sampling period and tentatively termed a mini-surge, was observed in 0%, 33%, 40%, and 60% of 30- to 33-, 34- to 37-, 38- to 42-, and 43- to 47-day-old rats, respectively. An overall increase in GnRH pulse frequency was observed in females displaying a mini-surge (9.0 pulses/5 h with mini-surge compared with 4.7 pulses/5 h with no mini-surge). The mini-surge itself, however, was associated with a late afternoon increase in GnRH pulse amplitude and not in pulse frequency. In adult proestrous rats, peak levels during the GnRH surge were an order of magnitude greater than those reached in pubertal animals. Our findings demonstrate that pubertal maturation in the female rat is associated with an acceleration of GnRH pulse generator activity and that later stages of pubertal maturation are characterized by the appearance of afternoon increases in GnRH release that may underlie previously reported mini-surges in LH. PMID- 11416014 TI - Identification, isolation, and cloning of growth hormone (GH)-inducible interscapular brown adipose complementary deoxyribonucleic acid from GH antagonist mice. AB - In a dwarf mouse line that expresses a GH antagonist, we found that the interscapular brown adipose tissue (iBAT) mass is significantly greater than that in nontransgenic littermates. We proposed that gene expression in iBAT may be up- or down-regulated by GH. To identify these genes, we employed the PCR-select subtraction approach to construct subtractive libraries from iBAT total RNAs. We have generated forward and reverse subtractive libraries. Clones were screened by differential hybridization and identified by BLAST similarity to expressed sequence tags and complementary DNA sequences. Four novel expressed sequence tags were isolated from the reverse subtractive library. Of them, clone 42, was further analyzed. It encodes a 2475-bp messenger RNA with an open reading frame of 346 amino acids. Northern blot analysis demonstrated two RNA isoforms (2.5 and 1.3 kb) in various tissues. Differential expression of both isoforms was verified in GH antagonist and nontransgenic mouse iBAT. BLAST searches suggested that clone 42 is highly homologous to a gene found in a human female fetal brain and a related gene found in a human pituitary tumor. PMID- 11416015 TI - Soy isoflavone supplements antagonize reproductive behavior and estrogen receptor alpha- and beta-dependent gene expression in the brain. AB - Epidemiological evidence suggests that isoflavone phytoestrogens may reduce the risk of cancer, osteoporosis, and heart disease, effects at least partially mediated by estrogen receptors alpha and beta (ERalpha and ERbeta). Because isoflavone dietary supplements are becoming increasingly popular and are frequently advertised as natural alternatives to estrogen replacement therapy, we have examined the effects of one of these supplements on estrogen-dependent behavior and ERalpha- and ERbeta-dependent gene expression in the brain. In the adult female rat brain, 17beta-estradiol treatment decreased ERbeta messenger RNA signal in the paraventricular nucleus by 41%, but supplement treatment resulted in a 27% increase. The regulation of ERbeta in the paraventricular nucleus is probably via an ERbeta-dependent mechanism. Similarly, in the ventromedial nucleus of the hypothalamus, supplement treatment diminished the estrogen dependent up-regulation of oxytocin receptor by 10.5%. The regulation of oxytocin receptor expression in the ventromedial nucleus of the hypothalamus is via an ERalpha-dependent mechanism. Supplement treatment also resulted in a significant decrease in receptive behavior in estrogen- and progesterone-primed females. The observed disruption of sexual receptivity by the isoflavone supplement is probably due to antiestrogenic effects observed in the brain. These results suggest that isoflavone phytoestrogens are antiestrogenic on both ERalpha- and ERbeta-dependent gene expression in the brain and estrogen-dependent behavior. PMID- 11416016 TI - Retarded liver growth in interleukin-6-deficient and tumor necrosis factor receptor-1-deficient mice. AB - The liver size in adult mammals is tightly regulated in relation to body weight, but the hormonal control of this is largely unknown. We investigated the roles of interleukin-6 (IL-6) and tumor necrosis factor (TNF) receptor-1 in the regulation of intact liver weight in adult mice. The relative liver wet and dry weights of older adult (5- to 10-month-old) IL-6 knockout (IL-6(-/-)) mice were decreased by 22-28%, and total contents of DNA and protein were decreased compared with those in age-matched wild-type mice. Weights of other visceral organs were unaffected. Older adult (6- to 8-month-old) TNF receptor-1 knockout (TNFR1(-/-)) mice displayed decreased relative liver weight. Treatment with a single injection of IL-6 increased liver wet and dry weights in IL-6(-/-) and wild-type mice, but not TNFR1(-/-) mice. Treatment with TNFalpha enhanced liver weight and DNA synthesis of nonparenchymal liver cells at 24 h in wild-type, but not IL-6(-/-), mice. At 48 h, TNFalpha induced DNA synthesis in nonparenchymal cells and hepatocytes of both wild-type and IL-6(-/-) mice. In conclusion, TNF receptor-1 stimulation and IL-6 production are both necessary for normal liver weight gain in older adult mice. The results of TNFalpha and IL-6 treatment further indicate that the effects of TNF receptor-1 and IL-6 depend on each other for full stimulation of liver growth. PMID- 11416017 TI - Expression and regulation of type II iodothyronine deiodinase in human thyroid gland. AB - We have studied the expression of type II iodothyronine deiodinase (DII) in human thyroid tumors and cultured human thyroid cells to elucidate the mechanisms involved in the regulation of DII expression in human thyroid gland. Three cases with hyperfunctioning thyroid adenoma, including a case that showed an activating mutation of G(s)alpha with a constitutive activation of cAMP production in cultured cells, and six cases with papillary thyroid carcinoma were analyzed in the present study. Free T(3) was increased, whereas free T(4) was within the normal range in all patients with hyperfunctioning thyroid adenoma. Thyroid tumor tissue and surrounding nontumor tissue were obtained at the time of surgery, and DII expression was compared between tumor tissue and nontumor tissue in each case. Northern analysis demonstrated the presence of DII messenger RNA (mRNA) approximately 7.5 kb in size in all of the tumor and nontumor tissues. DII mRNA and DII activity in hyperfunctioning thyroid adenoma were significantly increased compared with those in nontumor tissue in each case. In contrast, DII mRNA and DII activity in papillary thyroid carcinoma were decreased compared with those in nontumor tissue in each case. DII mRNA and DII activity in cultured human thyroid cells were significantly stimulated by TSH in a dose-dependent manner. The promoter activity of the human DII gene including the complete cAMP response element, transfected to cultured human thyroid cells, was stimulated by (Bu)(2)cAMP. In summary, these results suggest that DII expression in human thyroid gland is regulated at the transcriptional level through the TSH receptor G(s)alpha-cAMP regulatory cascade, which may be related to the increase in circulating T(3) level in patients with Graves' disease and hyperfunctioning thyroid adenoma. PMID- 11416018 TI - The proliferative and antiapoptotic actions of growth hormone and insulin-like growth factor-1 are mediated through distinct signaling pathways in the Pro-B Ba/F3 cell line. AB - Biological actions of GH can be direct or mediated through insulin-like growth factor I (IGF-I). In the interleukin-3 (IL-3)-dependent Ba/F3 cell line, IGF-I induces cell cycle entry and proliferation. Ba/F3 cells expressing the rat GH receptor (Ba/F3 GHR cells) have been shown to escape from apoptosis and to proliferate under GH stimulation. Using the Ba/F3 GHR cell model, we sought to dissect the signals elicited specifically by IGF-I or GH. In contrast to IGF-I or IL-3, GH is able to maintain cell cycle entry of Ba/F3 GHR cells cultured for 7 days in the absence of serum. The presence of IGF-I messenger RNA was not detected by RT-PCR, and by RIA, IGF-I was not found in culture medium of Ba/F3 GHR cells, unstimulated or stimulated by GH. Moreover, the addition of an anti IGF-I antibody that blocks IGF-I effects suggests that the actions of GH are not mediated by IGF-I, but appear to be direct. GH or IGF-I stimulation increased expression of cyclins A and D(1) with comparable kinetics, whereas expression of p21(waf1/cip1) seemed delayed in IGF-I-stimulated cells compared with that in GH stimulated cells. Contrary to GH or IL-3, IGF-I did not induce nuclear factor kappaB DNA-binding activity in Ba/F3 cells. Inhibition of nuclear factor-kappaB through expression of the mutant IkappaBalpha (A32/36) abrogated the GH-mediated survival signal, but did not result in alterations of the cell cycle in Ba/F3 GHR cells treated with IGF-I. Phosphatidylinositol 3-kinase was required for both survival and proliferative responses to IGF-I. Transfection of a dominant negative form of AKT (AH-AKT) resulted in suppression of IGF-I-mediated cell survival, but not of the antiapoptotic effect of GH in Ba/F3 GHR cells. Thus, GH and IGF-I are able to promote cell survival and proliferation through independent and different pathways in Ba/F3 cells. PMID- 11416019 TI - Cholesterol and hydroxycholesterol sulfotransferases: identification, distinction from dehydroepiandrosterone sulfotransferase, and differential tissue expression. AB - In humans, the biotransformation of cholesterol and its hydroxylated metabolites (oxysterols) by sulfonation is a fundamental process of great importance. Nevertheless, the sulfotransferase enzyme(s) that carries out this function has never been clearly identified. Cholesterol is a relatively poor substrate for the previously cloned hydroxysteroid sulfotransferase (HST), i.e. dehydroepiandrosterone (DHEA) sulfotransferase (HST1). Recently, cloning of a single human gene that encodes for two proteins related to HST1 was reported. These newly cloned sulfotransferases (HST2a and HST2b), while exhibiting sequence similarity to other members of the soluble sulfotransferase superfamily, also contain unique structural features. This latter aspect prompted an examination of their substrate specificity for comparison with HST1. Thus, HST1, HST2a, and HST2b were overexpressed as fusion proteins and purified. Furthermore, a novel procedure for the isolation of cholesterol and oxysterol sulfonates was developed that was used in association with HPLC to resolve specific sterol sulfonates. HST1 preferentially sulfonated DHEA and, to a lesser extent, oxysterols; whereas cholesterol was a negligible substrate. The reverse, however, was the case for the HST2 isoforms, particularly HST2b, which preferentially sulfonated cholesterol and oxysterols, in contrast to DHEA, which served as a poor substrate for this enzyme. RT-PCR analysis revealed distinct patterns of HST1, HST2a, and HST2b expression. It was particularly notable that both HST2 isoforms, but not HST1, were expressed in skin, a tissue where cholesterol sulfonation plays an important role in normal development of the skin barrier. In conclusion, substrate specificity and tissue distribution studies strongly suggest that HST2a and HST2b, in contrast to HST1, represent normal human cholesterol and oxysterol sulfotransferases. Furthermore, this study represents the first example of the sulfonation of oxysterols by a specific human HST. PMID- 11416020 TI - Induction of UO-44 gene expression by tamoxifen in the rat uterus and ovary. AB - A complementary DNA, uterine-ovarian-specific gene 44 (UO-44), has been isolated from tamoxifen-induced rat uterine complementary DNA library using differential display techniques. UO-44 transcripts are found to be abundant in the uterus and ovary. UO-44 gene expression in the uterus is strictly regulated by estrogens, tamoxifen, and GH, whereas the pure antiestrogen ICI 182780 is inhibitory. Treatment of ovariectomized rats and hypophysectomized rats with tamoxifen and GH, respectively, resulted in up-regulation of UO-44 expression in a dose dependent manner. In situ hybridization revealed that UO-44 gene expression was restricted to the luminal and glandular epithelial cells of the uterus and to granulosa cells of medium-size ovarian follicles. Transfection studies showed that UO-44 was a membrane-associated protein. Because estrogens, tamoxifen, and GH are stimulators of uterine luminal epithelial cell growth in vivo, UO-44 protein may serve as a mediator of the effect of these compounds in inducing epithelial proliferation and differentiation in these tissues. PMID- 11416021 TI - Lack of effect of protein deprivation-induced intrauterine growth retardation on behavior and corticosterone and growth hormone secretion in adult male rats: a long-term follow-up study. AB - To further define the neuroendocrine consequences of intrauterine growth retardation (IUGR), we have used a rat model of maternal protein restriction throughout pregnancy to examine the pattern of corticosterone and GH secretion under basal conditions and in response to psychological stress in male offspring at 4, 9, and 18 months of age. The findings were correlated with studies of behavioral activity. Despite a consistent reduction in birth weight and failure of catch-up growth, there were no significant differences in GH secretory profiles between IUGR and control rats at any age. We were unable to demonstrate a difference in the number, amplitude, length, or area of corticosterone secretory pulses between control and IUGR animals; although again, there was a significant decrease with age. The mean peak plasma concentration of corticosterone in response to a noise stress also declined with age but was unaffected by IUGR. There were no consistent, statistically significant differences in behavioral responses between normal control and IUGR animals or between groups of animals at different ages. These results do not, therefore, support the presence of major functional abnormalities in either GH or corticosterone secretory responses in adult male rats subjected to IUGR. PMID- 11416022 TI - Neurotensin gene expression increases during proestrus in the rostral medial preoptic nucleus: potential for direct communication with gonadotropin-releasing hormone neurons. AB - Neurotensin (NT)-containing neurons in the rostral portion of the medial preoptic nucleus (rMPN) of the brain may play a key role in regulating the pattern of secretion of GnRH, thereby influencing the reproductive cycle in females. The major goals of this study were to determine whether NT messenger RNA (mRNA) levels in the rMPN exhibit a unique pattern of expression in temporal association with the preovulatory LH surge and to assess whether NT neurons may communicate directly with GnRH neurons. We analyzed NT gene expression in rats using in situ hybridization over the day of proestrus and compared this with diestrous day 1. We also determined whether the high-affinity NT receptor (NT1) is expressed in GnRH neurons using dual-label in situ hybridization and whether this expression varies over the estrous cycle. We found that NT mRNA levels in the rMPN increase significantly on the day of proestrus, rising before the LH surge. No such change was detected on diestrous day 1, when the LH surge does not occur. Furthermore, we observed that a significant number of GnRH neurons coexpress NT1 mRNA and that the number of GnRH neurons expressing NT1 mRNA peaks on proestrus. Together with previous findings, our results suggest that increased expression of NT in the rMPN may directly stimulate GnRH neurons on proestrus, contributing to the LH surge. In addition, our results suggest that responsiveness of GnRH neurons to NT stimulation is enhanced on proestrus due to increased expression of NT receptors within GnRH neurons. PMID- 11416023 TI - Pregnancy-specific enhancement of agonist-stimulated ERK-1/2 signaling in uterine artery endothelial cells increases Ca(2+) sensitivity of endothelial nitric oxide synthase as well as cytosolic phospholipase A(2). AB - Uterine artery endothelial cells (UAEC) from pregnant ewes (P-UAEC) demonstrate generally enhanced ability to couple growth factor and G protein-coupled receptors to the ERK-1/2 signaling pathway and stimulate NO production independently of elevated [Ca(2+)]. Herein we investigate the signaling and vasodilator responses to ATP, an agonist that also elevates [Ca(2+)](i) in both NP and P-UAEC, to determine the relative importance of Ca(2+) vs. ERK-1/2 in the activation of eNOS. We observed in both NP-UAEC and P-UAEC that ATP acts through G protein-coupled P(2Y) receptors to activate phospholipase C and dose dependently elevate [Ca(2+)](i) independently of extracellular Ca(2+). The small reduction in the [Ca(2+)](i) response in NP vs. P-UAEC did not, however, account for the difference in NO production by P-UAEC>>NP-UAEC. ATP had no stimulatory effect on Akt phosphorylation but rapidly stimulated ERK-1/2 phosphorylation in P UAEC>>NP-UAEC in a manner that correlated with NO production. In both NP- and P UAEC, both ERK-1/2 and Ca(2+) were absolutely required for eNOS as well as cPLA(2) activation and the Ca(2+) sensitivity of eNOS was enhanced through the cytosolic [Ca(2+)](i) range in P-UAEC>>NP-UAEC. Thus ERK-1/2 may regulate the Ca(2+) sensitivity of eNOS to an even greater extent than is known to occur for cPLA(2). PMID- 11416024 TI - Leptin and tumor necrosis factor-alpha induce the tyrosine phosphorylation of signal transducer and activator of transcription proteins in the hypothalamus of normal rats in vivo. AB - Tumor necrosis factor-alpha (TNFalpha) reduces food intake and participates in the regulation of energy homeostasis. However, TNFalpha signaling in the brain and the potential interaction with leptin have not been investigated to date. Here we studied the tyrosine phosphorylation of STAT (signal transducer and activator of transcription) proteins in the hypothalamus of normal rats after iv injection of recombinant murine leptin or TNFalpha or coinjection of both cytokines. Immunoblot analysis of hypothalamic lysates with a phospho-specific STAT3 antibody showed a 6- to 7-fold stimulation of STAT3 tyrosine phosphorylation in response to both leptin and TNFalpha. Importantly, when coinjecting both cytokines, a remarkable synergistic activation (24-fold increase in STAT3 phosphorylation) could be detected. No other STAT proteins (STAT1, STAT5) were activated by leptin, whereas TNFalpha injection resulted in a dose dependent phosphorylation of hypothalamic STAT5. In contrast to its action in the brain, leptin was unable to produce STAT3 phosphorylation in the liver, either alone or in combination with TNFalpha. These data show that TNFalpha, independently of leptin, activates hypothalamic STAT signaling pathways and enhances leptin action at the level of STAT3. We therefore suggest that TNFalpha may represent a modulator of leptin action in the hypothalamus. PMID- 11416025 TI - Control of mouse cumulus cell-oocyte complex integrity before and after ovulation: plasminogen activator synthesis and matrix degradation. AB - During the preovulatory period, cumulus cells (CCs) form a hyaluronan-protein extracellular matrix (cumulus expansion) that positively influences oocyte fertilization. Degradation of this matrix and CC-oocyte complex (COC) dissociation occurs within a few hours of ovulation and parallels the aging of oocytes. Modulation of CC proteolytic activity by gonadotropins and oocyte soluble factors has been hypothesized to determine such cumulus matrix changes. In the present study, we investigated plasminogen activator (PA) synthesis by COCs during the expansion and disassembly processes. Our results show that the secretion of tissue type PA and urokinase type PA (uPA) by oocytes and CCs, respectively, does not change significantly during expansion but dramatically increases thereafter. Compact COCs were isolated from immature mice, primed 48 h earlier with 5 IU PMSGs, and were induced to expand in vitro with 100 ng/ml FSH in the presence of 1% FCS. Full expansion was achieved at 16 h, when hyaluronan synthesis ceased. Release of hyaluronan and CCs from the COC matrix began between 18 and 20 h of culture, which indicates that matrix degradation started at this time. PA activities in culture media were determined by SDS-PAGE, followed by a zymography at various time intervals between 4 and 32 h of culture. Secreted tissue type PA and uPA activity abruptly increased between 16 and 20 h after FSH stimulation. Slot blot hybridization of CC messenger RNA showed that uPA messenger RNA levels correlated with the increase in uPA activity. Similar temporal patterns of PA synthesis and matrix degradation were found in COCs induced to expand in vivo by injection of 5 IU human CG into PMSG-primed mice. Cultures of CCs, both in the presence and absence of oocytes, revealed that uPA synthesis is repressed in FSH-stimulated CCs by an oocyte-soluble factor for the first 16 h of culture, whereas CC responsiveness to this factor is lost thereafter. In conclusion, the data show that a sophisticated interplay between oocyte and CCs causes the two cell types to simultaneously secrete PA activity after ovulation. The fact that matrix degradation parallels PA production strongly supports the hypothesis that these enzymes may destabilize the expanded COC matrix. PMID- 11416026 TI - Cyclic adenosine 3',5'-monophosphate inhibits insulin-like growth factor I gene expression in rat glioma cell lines: evidence for regulation of transcription and messenger ribonucleic acid stability. AB - cAMP inhibits growth and stimulates differentiation in glioma cells. We examined the effect of cAMP on insulin-like growth factor I (IGF-I) gene expression in the C6 cell line, a rat glioma cell line previously reported to grow in response to autocrine IGF-I. cAMP potently inhibited IGF-I messenger RNA (mRNA) and peptide secretion in C6 cells, associated with an attenuation of DNA synthesis. Exogenous IGF-I peptide at least partially prevented the inhibition of DNA synthesis, suggesting that the reduction in IGF-I biosynthesis may contribute to the inhibitory effect of cAMP on C6 cell growth. cAMP also inhibited IGF-I mRNA in rat RG2 glioma cells, but not in three other nonglioma tumor cell lines. The nuclear IGF-I pre-mRNA level and the half-life of mature IGF-I mRNA were both reduced by cAMP in C6 cells, suggesting effects on gene transcription and mRNA stability. However, cAMP had no effect on the activities of IGF-I exon 1 promoter luciferase constructs. Protein synthesis inhibition partially reduced the inhibition of IGF-I mRNA by cAMP. Inhibition of cAMP-activated protein kinase A activity by H89 did not alter the inhibition of IGF-I gene expression in response to cAMP, suggesting that protein kinase A does not mediate the cAMP inhibitory effect on IGF-I gene expression. PMID- 11416027 TI - Gonadotropin regulation of NGFI-B messenger ribonucleic acid expression during ovarian follicle development in the rat. AB - NGFI-B is an immediate-early gene that encodes an orphan nuclear receptor. The present study was designed to examine the localization and gonadotropin regulation of NGFI-B expression in the rat ovary. Northern blot analysis of ovaries obtained from prepubertal rats revealed the increased expression of NGFI B during prepubertal development. Treatment of immature rats with PMSG, however, decreased ovarian NGFI-B expression. The major cell types expressing NGFI-B messenger RNA were thecal cells of follicles in different sizes. In contrast, treatment of PMSG-primed rats with human (h) CG resulted in the rapid and transient stimulation of ovarian NGFI-B messenger RNA, reaching a peak within 1 h. In situ hybridization analysis revealed that hCG treatment induced the expression of NGFI-B in granulosa cells of preovulatory follicles. Treatment of cultured preovulatory follicles in vitro with LH further confirmed the time- and dose-dependent stimulation of NGFI-B messenger RNA and protein. LH-stimulated NGFI-B expression in preovulatory follicles was abolished by alpha-amanitin, but was superinduced by cycloheximide. Furthermore, treatment of adult cycling rats with pentobarbital abolished NGFI-B expression on proestrus, and exogenous administration of hCG restored it, indicating the role of the preovulatory surge of LH in the stimulation of NGFI-B expression. These results demonstrate the cell type-specific expression and gonadotropin induction of NGFI-B in granulosa cells of preovulatory follicles and suggest a role for NGFI-B in the ovulatory process. PMID- 11416028 TI - Melanotrope cell plasticity: a key mechanism for the physiological adaptation to background color changes. AB - The intermediate lobe of the pituitary secretes the melanotropic hormone alpha MSH, which in amphibians plays a crucial role in skin color adaptation. It has been previously demonstrated that, in the frog Rana ridibunda, the intermediate lobe is composed of two distinct subpopulations of melanotrope cells that can be separated in vitro by using Percoll density gradients. These two melanotrope cell subsets, referred to as high-density (HD) and low-density (LD) cells, differ in their ultrastructural characteristics as well as in their biosynthetic and secretory activity. However, the specific, physiological role of the heterogeneity displayed by melanotrope cells remains elusive. In the present study, we investigated the effects of background color adaptation on melanotrope cell subpopulations. We found that adaptation of frogs to dark or white environment did not modify either the overall number of cells per intermediate lobe or the apoptotic and proliferation rates of melanotrope cells. On the other hand, adaptation of the animals to a white background significantly increased the proportion of hormone-storage HD cells and caused a concomitant decrease in that of LD cells (which exhibit higher levels of alpha-MSH release and POMC messenger RNA than HD cells). Conversely, after black-background adaptation the proportion of LD cells was markedly increased, suggesting that interconversion of HD cells to LD cells occurs during physiological activation of the intermediate lobe. In addition, black-background adaptation also enhanced alpha-MSH release by both cell subpopulations and increased inositol phosphate production in LD cells. These data indicate that, in frog, the proportions of the two melanotrope cell subsets undergo marked modifications during skin color adaptation, likely reflecting the occurrence of a secretory cell cycle whose dynamics are highly correlated to the hormonal demand imposed by the environment. PMID- 11416029 TI - Enhanced activity in parathyroid hormone-(1-14) and -(1-11): novel peptides for probing ligand-receptor interactions. AB - The amino-terminal portion of PTH is critical for PTH-1 receptor (P1Rc) activation. In exploring this component of the ligand receptor interaction, we recently showed that the agonist potency of the weakly active PTH-(1-14)NH(2) peptide can be enhanced by natural amino acid substitutions at several positions, including position 11 (normally leucine). Here we show that the potency of PTH-(1 14)NH(2) can be enhanced by using nonnatural amino acids that increase the length and polarizability of the position 11 side-chain. Thus, in LLC-PK(1) cells stably expressing high levels of the human P1Rc, [homoarginine([Har)(11)]PTH-(1-14)NH(2) was 30-fold more potent for cAMP production than was native PTH-(1-14)NH(2). Combining the homoarginine-11 substitution with other recently identified activity-enhancing substitutions yielded [Ala(3,12),Gln(10),Har(11),Trp(14)]PTH (1-14)NH(2), which was 1500-fold more potent than PTH-(1-14)NH(2) (EC(50) = 0.12 +/- 0.04 and 190 +/- 20 microM, respectively) and only 63-fold less potent than PTH-(1-34) (EC(50) = 1.9 +/- 0.5 nM). The even shorter analog [Ala(3),Gln(10),Har(11)]PTH-(1-11)NH(2) was also a full cAMP agonist (EC(50) = 3.1 +/- 1.5 microM). Receptor mutations at Phe(184) and Leu(187) located near the boundary of the amino-terminal domain and transmembrane domain-1 severely impaired responsiveness to the PTH-(1-11) analog. Overall, these studies demonstrate that PTH analogs of only 11 amino acids are sufficient for activation of the PTH-1 receptor through interaction with its juxtamembrane region. PMID- 11416030 TI - Activation of angiotensin II subtype 2 receptor induces catecholamine release in an extracellular Ca(2+)-dependent manner through a decrease of cyclic guanosine 3',5'-monophosphate production in cultured porcine adrenal medullary chromaffin Cells. AB - We have previously demonstrated that CGP 42112 (AT(2) agonist > or =1 nM) markedly reduces catecholamine biosynthesis through AT(2), which is the major angiotensin II (AngII) receptor subtype in cultured porcine chromaffin cells. Also, we have shown that CGP 42112 (> or =1 nM) induces a reduction in cGMP production in these cells. The present study showed that AngII reduced cGMP production via AT(2) in a manner similar to that found with CGP 42112. AngII (1 nM) significantly increased catecholamine secretion from cultured porcine adrenal medullary chromaffin cells. The stimulation was significantly inhibited by PD 123319 (AT(2) antagonist). The stimulation was moderately, but significantly, attenuated by CV-11974 (AT(1) antagonist, > or =10 nM), suggesting an involvement of AT(1). Moreover, CGP 42112 (> or =10 nM) markedly increased catecholamine release from these cells. The stimulation by CGP 42112 was abolished by PD 123319, whereas CV-11974 had no effect, indicating that this response is also mediated by AT(2). We further examined whether extracellular Ca(2+) is involved in the stimulatory effect of AT(2) on catecholamine secretion. Removal of external Ca(2+) significantly suppressed either AngII plus CV-11974 (100 nM; which simulates specific AT(2) stimulation) or CGP 42112- induced catecholamine secretion. AngII plus CV-11974 or CGP 42112 caused a sustained increase in intracellular Ca(2+) ([Ca(2+)](i)), as determined in fura-2-loaded chromaffin cells in an extracellular Ca(2+)-dependent manner. In the presence of EGTA, the subsequent addition of AngII with CV-11974 and CGP 42112 did not cause any increase in [Ca(2+)](i) levels. Consistent with this finding, CGP 42112 (10 nM to 1 microM) did not alter inositol triphosphate (IP(3)) production, a messenger for mobilization of Ca(2+) from intracellular storage sites. In addition, the intracellular Ca(2+) chelator 1,2-bis(2-amino-phenoxy)ethane-N,N,N',N'- tetraacetic acid acetoxymethylester (BAPTA) did not affect CGP 42112-induced catecholamine release. We tested whether a decrease in cGMP was the cause of the stimulatory effect of AT(2) on catecholamine secretion. Pretreatment with 8-bromo cGMP (1 mM) prevented the stimulatory effect of AngII plus CV-11974 and CGP 42112 on both catecholamine secretion and [Ca(2+)](i). When 8-bromo-cGMP was added after application of AngII plus CV-11974 or CGP 42112, [Ca(2+)](i) induced by these agents was gradually reduced toward the baseline values. Similarly, guanylin completely abolished the AngII- plus CV-11974-induced increase in both NE secretion and [Ca(2+)](i). The Ca(2+) channel blockers, nicardipine and omega conotoxin G VIA, at 1 microM in both cases, were also effective in inhibiting AT(2) stimulation-induced secretion. On the other hand, neither T-type voltage dependent Ca(2+) channel blockers, flunarizine, nor Ni(2+) affected catecholamine release caused by AT(2) stimulation. These findings demonstrate that AT(2) stimulation induces catecholamine secretion by mobilizing Ca(2+) through voltage dependent Ca(2+) channels without affecting intracellular pools and that these effects could be mediated by a decrease in cGMP production. PMID- 11416031 TI - High salt intake increases uroguanylin expression in mouse kidney. AB - The intestinal peptides, guanylin and uroguanylin, may have an important role in the endocrine control of renal function. Both peptides and their receptor, guanylyl cyclase C (GC-C), are also expressed within the kidney, suggesting that they may act locally in an autocrine/paracrine fashion. However, their physiological regulation within the kidney has not been studied. To begin to address this issue, we evaluated the distribution of uroguanylin and guanylin messenger RNA (mRNA) in the mouse nephron and the regulation of renal expression by changes in dietary salt/water intake. Expression was determined in 1) wild type mice, 2) two strains of receptor-guanylyl cyclase-deficient mice (ANP receptor-deficient, GC-A-/-, and GC-C-deficient mice); and 3) cultured renal epithelial (M-1) cells, by RT-PCR, Northern blotting and immunocytochemistry. Renal uroguanylin messenger RNA expression was higher than guanylin and had a different distribution pattern, with highest levels in the proximal tubules, whereas guanylin was mainly expressed in the collecting ducts. Uroguanylin expression was significantly lower in GC-C-/- mice than in GC-A-/- and wild types, suggesting that absence of a receptor was able to down-regulate ligand expression. Salt-loading (1% NaCl in drinking water) increased uroguanylin-mRNA expression by >1.8-fold but had no effect on guanylin expression. Uroguanylin but not guanylin transcripts were detected in M-1 cells and increased in response to hypertonic media (+NaCl or mannitol). Our results indicate that high-salt intake increases uroguanylin but not guanylin expression in the mouse kidney. The synthesis of these peptides by tubular epithelium may contribute to the local control of renal function and its adaptation to dietary salt. PMID- 11416032 TI - Activation of the insulin-like growth factor 1 signaling pathway by the antiapoptotic agents aurintricarboxylic acid and evans blue. AB - Aurintricarboxylic acid (ATA), an endonuclease inhibitor, prevents the death of a variety of cell types in culture. Previously we have shown that ATA, similar to insulin-like growth factor I (IGF-I), protected MCF-7 cells against apoptotic death induced by the protein synthesis inhibitor cycloheximide. Here we show that ATA and a polysulfonated aromatic compound, Evans blue (EB), similar to IGF-I, promote survival and increase proliferation of MCF-7 cells in serum-free culture medium. This may suggest a common signaling pathway shared by the aromatic polyanions and IGF-I. Therefore, the ability of these aromatic compounds to activate the signal transduction pathway of IGF-I was examined. We found that ATA and EB mimicked the IGF-I effect on tyrosine phosphorylation of the IGF-I receptor (IGF-IR) and its major substrates, insulin receptor substrate-1 (IRS-1) and IRS-2; induced the association of these substrates with phosphatidylinositol 3-kinase and Grb2; and activated Akt kinase and p42/p44 mitogen-activated protein kinases. ATA and EB competed for IGF-I binding to the IGF-IR. ATA was found to be selective for the IGF-IR, whereas EB also activated the insulin receptor. Upon fractionation of commercial ATA by size exclusion chromatography, we found that fractions that enhanced the intensity of tyrosyl-phosphorylated IRS-1/IRS-2 also increased the survival of MCF-7 cells in the presence of cycloheximide, whereas fractions devoid of IRS phosphorylation activity had no survival ability. Taken together, these results suggest that the survival/proliferation-promoting effects of ATA and EB in MCF-7 cells are transduced via the IGF-IR signaling pathway. PMID- 11416033 TI - Luteinizing hormone secretion from wild-type and progesterone receptor knockout mouse anterior pituitary cells. AB - The progesterone receptor (PR) has a central role in the hypothalamo-pituitary events culminating in the preovulatory LH surge, and mice with genetically ablated PR provide a model for dissecting cellular pathways subserving this role. The aims of this study were to determine 1) whether the GnRH self-priming response and acute progesterone augmentation of secretagogue-stimulated LH secretion are present in cultured wild-type (WT) mouse pituitary cells, and 2) whether the PR is essential for self-priming by comparing the responses in PR knockout (PRKO) cells. Pituitary cells from ovariectomized WT or PRKO mice cultured +/- 17beta-estradiol (E(2)) for 3 days were challenged with hourly pulses of 1 nM GnRH or 54 mM K(+). A background of E(2) had no effect on the initial LH secretory response for either WT or PRKO cells. However, for subsequent GnRH pulses, E(2) was permissive for the GnRH self-priming response in WT cells. PRKO cells exhibited a blunted GnRH self-priming response. Exposure to progesterone for 90 min before secretagogue stimulation resulted in a modest (1.5 fold) augmentation of the LH response to GnRH but not K(+) pulses in WT cells; progesterone had no effect in PRKO cells. Unlike in the rat, the PR antagonists RU486 or ZK98299 failed to prevent potentiation of LH secretory responses to multiple GnRH pulses in WT cells. Although RU486 blocked progesterone augmentation of the initial GnRH pulse, it was ineffective in blocking progesterone's action after multiple GnRH pulses. In WT cells, 8- bromo-cAMP (8 Br-cAMP) was able to substitute for the GnRH priming pulse; 8-Br-cAMP also augmented GnRH-stimulated secretion in PRKO cells but less effectively. 8-Br-cAMP augmented K(+)-stimulated LH secretion in WT and PRKO cells equally. These results suggest that, although mouse gonadotropes show GnRH self-priming, they have adapted strategies different than rat cells for amplifying the GnRH signal as shown by the residual self-priming in PRKO cells, the modest or absent augmentation by acute progesterone of GnRH- or K(+)-stimulated secretion in WT cells, and the reduced ability of PR antagonists to interfere with GnRH self priming and progesterone augmentation. We speculate that the adaptations could involve, at least in part, differences in the ratio of PR isoforms. PMID- 11416034 TI - Regulation of steroidogenic acute regulatory protein and luteinizing hormone receptor messenger ribonucleic acid in hen granulosa cells. AB - The regulation of steroidogenic acute regulatory protein (StAR) in vitro by gonadotropins was investigated in granulosa cells from prehierarchal and preovulatory hen follicles. Basal levels of StAR messenger RNA (mRNA) in undifferentiated granulosa cells from prehierarchal (6- to 8-mm) follicles were consistently low, but detectable, and were significantly increased by treatment with 8-bromo-cAMP and FSH (but not LH) within 3-6 h of culture. After 20 h of culture, 8-bromo-cAMP, FSH, and LH each increased StAR mRNA levels above those in control cultured cells, and the delayed response to LH treatment was associated with increased levels of LH receptor (LH-R) mRNA. On the other hand, inhibition of mitogen-activated protein (MAP) kinase signaling, using the MAP kinase kinase inhibitors U0126 and PD98059, in the presence of FSH further increased StAR mRNA and protein levels, LH-R mRNA levels, and progesterone synthesis compared with those in cells cultured with FSH alone. The highest basal expression of StAR mRNA during follicle development was found in granulosa from the largest (F1) preovulatory follicle, with comparatively lower levels in granulosa from less mature (F2 plus F3) preovulatory follicles. Treatment with LH rapidly increased StAR mRNA and protein (but not LH-R mRNA) expression in cultures of F1 granulosa and in combined F2 plus F3 granulosa within 3 h, although the magnitude of stimulation was greater in F2 plus F3 granulosa. Compared with results from granulosa cells from prehierarchal follicles cultured for 20 h, inhibition of MAP kinase signaling in the presence of LH for 1 h failed to further enhance levels of StAR or LH-R expression or progesterone production in F2 plus F3 follicle granulosa compared with the effect of LH treatment alone. These results demonstrate that StAR expression in the hen ovary is up-regulated by gonadotropins at least in part via cAMP signaling. The ability of MAP kinase kinase inhibitors to potentiate gonadotropin-induced StAR and LH-R expression plus progesterone synthesis in prehierarchal follicle granulosa cells in vitro suggests that inhibition of paracrine or autocrine factor-mediated MAP kinase signaling in vivo may be a prerequisite for the full potentiation of granulosa cell steroidogenesis that occurs after recruitment into the preovulatory hierarchy. Finally, these results fail to support a role for MAP kinase signaling in acutely modulating LH-mediated StAR expression or progesterone production in hierarchal follicles, such as occurs during the preovulatory surge of progesterone. PMID- 11416035 TI - Transcriptional suppression of type 1 angiotensin II receptor gene expression by peroxisome proliferator-activated receptor-gamma in vascular smooth muscle cells. AB - Angiotensin (A) II plays a critical role in vascular remodeling, and its action is mediated by type 1 AII receptor (AT1R). Recently, 15-deoxy-(Delta)(12,14) prostaglandin J(2) and thiazolidinediones have been shown to be ligands for peroxisome proliferator-activated receptor (PPAR)-gamma and activate PPAR-gamma. In the present work, we have studied the effect of PPAR-gamma on AT1R expression in rat vascular smooth muscle cells (VSMCs). We observed that: 1) endogenous AT1R expression was significantly decreased by PPAR-gamma ligands both at messenger RNA and protein levels, whereas AT1R messenger RNA stability was not affected; 2) AII-induced increase of (3)H-thymidine incorporation into VSMCs was inhibited by PPAR-gamma ligands; 3) rat AT1R gene promoter activity was significantly suppressed by PPAR-gamma ligands, and PPAR-gamma overexpression further suppressed the promoter activity; 4) transcriptional analyses using AT1R gene promoter mutants revealed that a GC-box-related sequence within the -58/-34 region of the AT1R gene promoter was responsible for the suppression; 5) Sp1 overexpression stimulated AT1R gene transcription via the GC-box-related sequence, which was inhibited by additional PPAR-gamma overexpression; 6) electrophoretic mobility shift assay suggested that Sp1 could bind to the GC-box related sequence whereas PPAR-gamma could not; 7) antibody supershift experiments using VSMC nuclear extracts revealed that protein-DNA complexes formed on the GC box-related sequence, which were decreased by PPAR-gamma coincubation, were mostly composed of Sp1; and 8) glutathione S-transferase pull-down assay revealed a direct interaction between PPAR-gamma and Sp1. Taken together, it is suggested that activated PPAR-gamma suppresses AT1R gene at a transcriptional level by inhibiting Sp1 via a protein-protein interaction. PPAR-gamma ligands, thus, may inhibit AII-induced cell growth and hypertrophy in VSMCs by AT1R expression suppression and possibly be beneficial for treatment of diabetic patients with hypertension and atherosclerosis. PMID- 11416036 TI - Targeted inactivation of the 25-hydroxyvitamin D(3)-1(alpha)-hydroxylase gene (CYP27B1) creates an animal model of pseudovitamin D-deficiency rickets. AB - Pseudovitamin D-deficiency rickets is caused by mutations in the cytochrome P450 enzyme, 25-hydroxyvitamin D(3)-1alpha-hydroxylase (1alpha-OHase). Patients with the disease exhibit growth retardation, rickets, and osteomalacia. Serum biochemistry is characterized by hypocalcemia, secondary hyperparathyroidism, and undetectable levels of 1alpha,25-dihydroxyvitamin D(3). We have inactivated the 1alpha-OHase gene in mice after homologous recombination in embryonic stem cells. Serum analysis of homozygous mutant animals confirmed that they were hypocalcemic, hypophosphatemic, hyperparathyroidic, and that they had undetectable 1alpha,25-dihydroxyvitamin D(3). Histological analysis of the bones from 3-week-old mutant animals confirmed the evidence of rickets. At the age of 8 weeks, femurs from 1alpha-OHase-ablated mice present a severe disorganization in the architecture of the growth plate and marked osteomalacia. These results show that we have successfully inactivated the 1alpha-OHase gene in mice and established a valid animal model of pseudovitamin D-deficiency rickets. PMID- 11416037 TI - Interferon-gamma modulates prolactin and tissue factor expression in differentiating human endometrial stromal cells. AB - Cytokines such as interferon-gamma (IFNgamma) released by resident uterine immune cells are thought to influence the expression of differentiated function in the human endometrium. Decidualization of the stromal cell compartment is confined to the superficial endometrial layer in the nonpregnant uterus. To explore the molecular mechanism underlying the spatial expression of the decidual phenotype, the effect of IFNgamma on the induction of two well characterized markers of endometrial stromal (ES) cell differentiation, PRL and tissue factor (TF), has been investigated. IFNgamma antagonizes cAMP-mediated PRL protein and messenger RNA expression in primary ES cell cultures through inhibition of decidual PRL promoter activity. In parallel, IFNgamma stimulates Stat-1 (signal transducer and activator of transcription-1) expression, phosphorylation, and translocation to the nucleus. Exogenously expressed Stat-1 potently represses decidual PRL promoter activation, indicating the potential for the inhibitory effects of IFNgamma to be mediated by Stat-1. We demonstrate that although the coactivator CREB-binding protein/p300 is essential for decidual PRL transcription, this coactivator does not appear to be the target for IFNgamma-mediated repression. By contrast, IFNgamma has little effect on cAMP-mediated TF expression, but induces TF in ES cells not exposed to a decidualizing stimulus. This suggested that in vivo TF expression may not be restricted to decidualizing cells of the superficial layer and was confirmed by immunohistochemical analysis demonstrating intense TF staining in the basal stromal compartment during the regeneration phase of the cycle. The differential sensitivity of decidualization-associated genes to IFNgamma illustrates its potential role as a selective biological response modifier that influences regional function within the endometrium. PMID- 11416038 TI - Dystroglycan is present in rat thyroid and rat thyroid cells and responds to thyrotropin. AB - Dystroglycan is a high affinity laminin-binding glycoprotein originally described as a member of the dystrophin-associated glycoprotein complex in muscle. We have demonstrated the presence of dystroglycan in the thyroid using immunocytochemistry, immunoblots, ligand binding assays, and relative quantitative RT-PCR. In intact rat thyroid glands, antibodies against the alpha (extracellular, laminin-binding subunit) and beta (cytoplasmic/membrane bound) portions of the dystroglycan protein reacted at basolateral membranes where they colocalized with laminin. Western-blotted protein from the Fischer rat thyroid cell line FRTL-5 reacted with both the alpha- and beta-dystroglycan antibodies. The alpha-dystroglycan-reactive band colocalized with laminin-binding activity, and the protein and binding activity were decreased by TSH. In contrast, in the culture medium of these cells, alpha-dystroglycan was increased by TSH. The beta dystroglycan antibody recognized the full-length 43-kDa band and an approximately 30-kDa truncated form. The truncated form was reduced in cells cultured with TSH, whereas the full-length form was not significantly diminished by TSH. Immunofluorescence of FRTL-5 cells in the absence of TSH showed a colocalization of dystroglycan and laminin. This was disrupted by the addition of TSH and was correlated to morphological changes. PCR amplification of complementary DNA with primer pairs from alpha- and beta-dystroglycan produced appropriately sized bands, whose sequence had identical protein-coding sequences and more than 96% nucleotide homology to mouse dystroglycan sequences. Relative quantitative RT-PCR of beta-dystroglycan messenger RNA showed reduced expression in cells cultured with TSH. We conclude that dystroglycan is present in rat thyroid and in FRTL5 rat thyroid cells and that TSH reduces its expression. PMID- 11416039 TI - Microarray analysis of the in vivo effects of hypophysectomy and growth hormone treatment on gene expression in the rat. AB - Complementary DNA microarrays containing 3000 different rat genes were used to study the consequences of severe hormonal deficiency (hypophysectomy) on the gene expression patterns in heart, liver, and kidney. Hybridization signals were seen from a majority of the arrayed complementary DNAs; nonetheless, tissue-specific expression patterns could be delineated. Hypophysectomy affected the expression of genes involved in a variety of cellular functions. Between 16-29% of the detected transcripts from each tissue changed expression level as a reaction to this condition. Chronic treatment of hypophysectomized animals with human GH also caused significant changes in gene expression patterns. The study confirms previous knowledge concerning certain gene expression changes in the above mentioned situations and provides new information regarding hypophysectomy and chronic human GH effects in the rat. Furthermore, we have identified several new genes that respond to GH treatment. Our results represent a first step toward a more global understanding of gene expression changes in states of hormonal deficiency. PMID- 11416040 TI - Quantitative analysis of estrogen receptor proteins in rat mammary gland. AB - Estrogen receptor alpha and beta proteins (ERalpha and ERbeta) at various stages of development of the rat mammary gland were quantified by Western blotting. ERalpha and ERbeta recombinant proteins were used as standards, and their molar concentrations were measured by ligand binding assays. In 3-week-old pregnant, lactating, and postlactating rats the ERalpha content ranged from 0.30-1.55 fmol/microg total protein (mean values). The ERbeta content of the same samples ranged between 1.06-7.50 fmol/microg total protein. At every developmental stage, the ERbeta content of the mammary gland was higher than that of ERalpha. When receptor levels were normalized against beta-actin, it was evident that ER expression changed during development, with maximum expression of both receptors during the lactation period. With an antibody raised against the 18-amino acid insert of the ERbeta variant, originally called ERbeta2 but named ERbetains in this paper, Western blots revealed that ERbetains protein was up-regulated during the lactation period. RT-PCR showed that the levels of messenger RNA of ERbetains paralleled those of the protein. Double immunohistochemical staining with anti ERalpha and anti-ERbetains antibodies revealed that ERbetains protein colocalized with ERalpha in 70-80% of the ERalpha-expressing epithelial cells during lactation and with 30% of these cells during pregnancy. These observations indicate that expression of ERbetains is regulated not only quantitatively, but also with regard to its cellular distribution. As ERbetains acts as the dominant repressor of ERalpha, we suggest that its coexpression with ERalpha quenches ERalpha function and may be one of the factors that contribute to the previously described insensitivity of the mammary gland to estrogens during lactation. PMID- 11416041 TI - Regulation of prostaglandin-endoperoxide synthase 2 messenger ribonucleic acid expression in mouse granulosa cells during ovulation. AB - Normal ovulation in mice requires PG-endoperoxide synthase 2 (cyclooxygenase-2; COX-2) expression. This study examined the role of the oocyte and other factors in regulating steady state levels of COX-2 messenger RNA (mRNA) in granulosa cells. Multiphasic changes in the expression pattern of COX-2 mRNA were found, with peaks of expression 4 and 12 h after hCG treatment. Changes in relative expression levels in cumulus cells and mural granulosa cells occurred over time, with similar mRNA levels at 4 h, but higher levels in cumulus cells compared with mural granulosa cells at 8 and 12 h post-hCG. In cultured mural granulosa cells, LH, FSH, and oocytes promoted COX-2 mRNA expression concurrent with the first expression peak in vivo. At the same time, FSH, but not LH, treatment of cultured cumulus-oocyte complexes (COC) promoted COX-2 mRNA expression in cumulus cells. This response of cumulus cells to FSH treatment was largely dependent on the presence of either fully grown germinal vesicle stage or maturing oocytes, but not growing oocytes. At 8 h, COX-2 mRNA expression in FSH-stimulated COC was lower than at 4 h; however, oocyte coculture promoted COX-2 mRNA expression in cumulus cells. No second peak in expression occurred in cultured COC. However, coculture of COC with follicle walls promoted COX-2 mRNA expression in cumulus cells 12 h post-hCG; an effect augmented by oocytes. Therefore, the oocyte resident within ovulatory follicles produces a factor(s) that promotes expression of COX-2 mRNA by cumulus cells and possibly by mural granulosa cells. Thus, the oocyte probably plays an important role in promoting ovulation. However, the multiphasic changes in the pattern of COX-2 expression appear orchestrated by non oocyte-derived factors. PMID- 11416042 TI - Comparative analysis of pharmacologic and/or genetic disruption of cyclooxygenase 1 and cyclooxygenase-2 function in female reproduction in mice. AB - Cyclooxygenase (COX)-derived prostaglandins are critical in female reproduction. Gene targeting studies show that ovulation, fertilization, implantation, and decidualization are defective in COX-2 deficient mice. We used genetic and pharmacologic approaches to perturb COX function and examine the differential and synergistic effects of inhibition of COX-1, COX-2, or of both isoforms on reproductive outcomes during early pregnancy in mice. The results demonstrate that simultaneous inhibition of COX-1 and COX-2 produces more severe effects on early pregnancy events than inhibition of either isoform alone. The effects of pharmacological inhibition of COX-2 on female reproductive functions were less severe than the null mutation of the COX-2 gene. A combined approach showed that COX-2 inhibition in COX-1(-/-) mice induced complete reproductive failure, suggesting a lack of alternative sources of prostaglandin synthesis. This investigation raises caution regarding the indiscriminate use of COX inhibitors and shows for the first time the distinct and overlapping pathways of the cyclooxygenase systems in female reproduction. PMID- 11416043 TI - A novel potent antagonist of peroxisome proliferator-activated receptor gamma blocks adipocyte differentiation but does not revert the phenotype of terminally differentiated adipocytes. AB - The antidiabetic thiazolidinediones, which include troglitazone and rosiglitazone, are ligands for the nuclear receptor peroxisome proliferator activated receptor gamma (PPARgamma). Their antihyperglycemic effects seem to be linked to the regulation of PPARgamma-responsive genes. Here, we report the characterization of a specific PPARgamma antagonist that blocks several of the biological activities of the PPARgamma agonist rosiglitazone. PD068235 inhibited rosiglitazone-dependent PPARgamma transcriptional activity with an IC(50) of 0.8 microM and rosiglitazone-stimulated in vitro coactivator association. The role of PPARgamma in the initiation of differentiation is well documented. In this study, we used PD068235 as a tool to evaluate the functional role of PPARgamma in the maintenance of the terminally differentiated state. Treatment of confluent, growth-arrested 3T3-L1 preadipocytes with PD068235 blocked adipocyte differentiation induced by the standard adipogenic hormonal mixture (insulin/dexamethasone/isobutylmethylxanthin) and fully antagonized rosiglitazone induced adipogenesis. In contrast, long-term treatment of terminally differentiated 3T3-L1 adipocytes with PD068235 did not induce any obvious morphological changes and had no effect on basal lipolysis rates. In addition, in fully differentiated adipocytes PD068235 did not alter the basal expression of PPARgamma target genes aP2 and CAP, but it effectively blocked rosiglitazone induced expression of both genes. These results suggest that in terminally differentiated adipocytes, the PPARgamma activity is minimal and may not be required for the maintenance of PPARgamma target gene expression. PMID- 11416044 TI - Fibronectin and the alpha(5)beta(1) integrin are under developmental and ovarian steroid regulation in the normal mouse mammary gland. AB - Extracellular matrix (ECM) proteins have been shown to regulate mammary epithelial cell proliferation, differentiation, and apoptosis in vitro. However, little is known about the hormonal regulation and functional role of ECM proteins and integrins during mammary gland development in vivo. We examined the temporal and spatial localization and hormone regulation of collagen I, collagen IV, laminin, and fibronectin. Among these ECM proteins only fibronectin changed appreciably. Fibronectin levels increased 3-fold between the onset of puberty and sexual maturity, remaining high during pregnancy and lactation. This increase occurred specifically in the epithelial basement membrane. Fibronectin was decreased 70% by ovariectomy and increased 1.5- and 2-fold by estrogen or estrogen plus progesterone treatment, respectively. The fibronectin-specific integrin, alpha(5)beta(1), was localized in myoepithelial cells; it increased 2.2 fold between puberty and sexual maturity and decreased in late pregnancy and lactation. The basal localization of alpha(5)beta(1) was notably increased in pubertal and adult virgin mice. alpha(5)beta(1) concentrations decreased 40-50% after ovariectomy in pubertal and adult mice, which was reversed by estrogen plus progesterone treatment in adult mice. The high basal expression of alpha(5)beta(1) during active proliferation and the low expression in nonproliferating and lactating glands indicate that fibronectin signaling may be required for hormone-dependent proliferation in the mammary gland. PMID- 11416045 TI - Hydroxylases involved in vitamin D metabolism are differentially expressed in murine embryonic kidney: application of whole mount in situ hybridization. AB - In this study we examined the expression of 25-hydroxyvitamin D-1alpha hydroxylase (1alpha-hydroxylase) and 25-hydroxyvitamin D-24-hydroxylase (24 hydroxylase) by RT-PCR and whole mount in situ hybridization using organ culture of kidney taken from mouse embryo. First, the kidneys of mouse embryo at 11.5 17.5 days gestation were cultured in the presence or absence of forskolin and 1,25-dihydroxyvitamin D(3) [1alpha,25-(OH)(2)D(3)]. Forskolin and 1alpha,25 (OH)(2)D(3) induced the expression of 1alpha-hydroxylase and 24-hydroxylase, respectively, in a dose- and time-dependent manner. In the absence of stimulants, the expression of 1alpha-hydroxylase and 24-hydroxylase was detected from days 13.5-17.5 gestation. The expression of vitamin D receptor and megalin was detected from days 13.5 and 11.5, respectively. Next, signals for the expression of either 1alpha-hydroxylase or 24-hydroxylase were detected by whole mount in situ hybridization in kidney explants taken from embryo at 15.5 days gestation after the appropriate stimulation. However, the localization of signals differed between the two enzymes; 1alpha-hydroxylase messenger RNA was expressed in the inner area of the kidney explants, whereas 24-hydroxylase messenger RNA was expressed in the surface area. The expression of both hydroxylases was restricted to the epithelium of developing renal tubules. The pattern of megalin expression was similar to that of 1alpha-hydroxylase expression. To confirm the difference in distribution of 1alpha-hydroxylase and 24-hydroxylase transcripts, the explants were hybridized with probes for both 1alpha-hydroxylase and 24 hydroxylase using double labeling techniques after simultaneous stimulation with forskolin and 1alpha,25-(OH)(2)D(3), resulting in the detection at different locations of positive signals for the two enzymes. These results suggest that the expression of 1alpha-hydroxylase is induced in a distinct epithelium of renal tubules from that of 24-hydroxylase even at the early stage of kidney development before glomerulogenesis. PMID- 11416046 TI - Isolation and characterization of a novel member of the relaxin/insulin family from the testis of the frog Rana esculenta. AB - A complementary DNA (cDNA) encoding a frog relaxin/insulin member family (fRLX) from testis cDNA library was isolated and characterized. The fRLX cDNA predicted a 155-amino acid protein with a low homology to mammalian RLF and relaxin. Northern blot analysis revealed a single transcript expressed in the interstitial compartment, RT-PCR, evidenced that fRLX is expressed at low levels in the oviduct and ovary too. The predicted mature fRLX protein, composed of the signal peptide, B, C, and A domains, has conserved amino acid sequences in the characteristic functional domains. A different expression of the transcript was found during the frog reproductive cycle, with a peak in Spring. After administration of ethane dimethane sulfonate, by in situ hybridization, fRLX messenger RNA disappeared from the interstitial compartment and reappeared again at the time of generating of a new population of Leydig cells (LC), strongly indicating that LC are the interstitial cell type expressing fRLX. Preliminary results obtained by in situ hybridization, performed on testis of hypophysectomized frogs evidenced a pituitary control of fRLX expression. This study is the first cloning of a relaxin/insulin family member in a nonmammalian vertebrate. In addition, because fRLX expression changes during the annual cycle suggesting its involvement in spermatogenesis, fRLX may be considered a new marker for the study of spermatogenesis in the Rana esculenta. PMID- 11416047 TI - A novel function of prolactin-releasing peptide in the control of growth hormone via secretion of somatostatin from the hypothalamus. AB - The present study examined a novel function of PRL-releasing peptide (PrRP) on the neuroendocrine. PrRP-immunoreactive nerve fibers and nerve terminals were located in the vicinity of the somatostatin (SOM)-neurons in the hypothalamic periventricular nucleus (PerVN). Immuno-electron microscopy revealed that PrRP immunoreactive nerve terminals made synaptic contacts with nonimmunoreactive neuronal elements in the PerVN. Intracerebroventricular (icv) administration of PrRP induced immediate early gene, NGFI-A, in SOM-neurons in the PerVN. Double labeling in situ hybridization showed that some parts of SOM-neurons in the PerVN expressed PrRP receptor messenger RNA. Therefore, some parts of SOM-neurons in the PerVN are considered to be directly innervated by PrRP via PrRP receptor. In addition to the above morphological characteristics, icv administration of PrRP decreased plasma GH levels. Such inhibitory effects of PrRP on the secretion of GH from the anterior pituitary were diminished by depletion or neutralization of SOM. From these findings it was strongly suggested that SOM-neurons respond to PrRP and secrete SOM into the portal vessels and thus inhibit GH secretion from the anterior pituitary. PMID- 11416048 TI - Angiogenesis during follicular development in the primate and its inhibition by treatment with truncated Flt-1-Fc (vascular endothelial growth factor Trap(A40)). AB - The aims of this study were to 1) quantify changes in angiogenesis during follicular growth in a primate model; 2) investigate the molecular regulation using in situ hybridization of vascular endothelial growth factor (VEGF), its receptor, Flt-1, the angiopoietins (Ang-1 and Ang-2), and their receptor, Tie-2; 3) elucidate the role of VEGF in follicular angiogenesis by blocking its action by treatment with a soluble truncated form of the Flt-1 receptor, (VEGF Trap(A40)). Changes in angiogenesis were quantified using bromodeoxyuridine to obtain a proliferation index, and CD31 immunocytochemistry to visualize endothelial cell area. Percentage of proliferating endothelial cells was calculated by double labeling for bromodeoxyuridine and CD31. Vascularization was first observed in follicles containing four granulosa cell layers. A significant increase in proliferation in the thecal layer was observed from the early to late secondary stage, and dual staining showed that 25% of proliferating cells were of endothelial cell origin. VEGF messenger RNA (mRNA) was expressed in granulosa cells with an increase of grain density from late secondary to tertiary follicles. Ang-1 was weakly expressed in the theca of tertiary follicles. Ang-2 mRNA was not detected in any follicles. The mRNA for the Flt-1 and Tie-2 receptors was localized in endothelial cells of the theca. Unexpectedly, Tie-2 mRNA was also found in granulosa cells of early follicular stages and its translation was confirmed by immunocytochemistry. VEGF trap treatment for 3 days resulted in an 87% decrease of proliferation in the theca of secondary and tertiary follicles, a reduction in endothelial cell area and a marked decline in Flt-1 mRNA expression. Granulosa cell proliferation also decreased. These results show that onset and establishment of the follicle vasculature takes place early during follicular development. The ability of VEGF trap treatment to severely restrict follicular angiogenesis establishes that VEGF is the major regulator of this process in the primate ovary. PMID- 11416049 TI - Persistent synchronized oscillations in prolactin gene promoter activity in living pituitary cells. AB - PRL gene expression in the anterior pituitary gland responds rapidly to different hormonal signals. We have investigated the long-term timing of transcriptional activation from the PRL, GH, and cytomegalovirus promoters in response to different stimulus duration, using real-time imaging of luciferase expression in living stably transfected GH3 cells. Long-term stimulation of serum-starved cells with 50% serum induced a homogeneous rise in PRL promoter activity, with subsequent heterogeneous fluctuations in luciferase activity in individual cells. When cells were subjected to a 2-h pulse of 50% serum, followed by serum-free medium, there were long-term (approximately 50 h) synchronized, homogeneous oscillations in PRL promoter activity. This response was PRL-specific, because in GH3 cells expressing luciferase from the GH or cytomegalovirus promoters, a serum pulse elicited no oscillations in luciferase expression after an initial transient response to serum. The PRL promoter may therefore be a template for an unstable transcription complex subject to stochastic regulation, allowing an oscillatory transcriptional response to physiological signals. This suggests that precise timing and coordination of cell responses to different signal-duration may represent a novel mechanism for coordinating long-term dynamic changes in transcription in cell populations. PMID- 11416050 TI - Fish growth hormone receptor: molecular characterization of two membrane-anchored forms. AB - A RT-PCR approach was used to clone and sequence the full-length growth hormone receptor (GHR) of a teleost fish, the turbot (Scophthalmus maximus). Total liver RNA was amplified by RT-PCR with degenerate primers designed in extracellular and cytoplasmic regions, and a single DNA fragment of 1100 bp was obtained. The entire coding region was obtained by 5' and 3' RACE assays, and comprises an open reading frame of 633 amino acids. This sequence shows the characteristic motifs of the class I cytokine receptor superfamily, and its amino acid identity with mammalian, avian, reptilian and amphibian GHRs is 32-36%. The 3' RACE also revealed the occurrence of an alternate messenger encoding a membrane-anchored truncated receptor, which could facilitate the production of GH-binding protein in fish species. This report represents the first data on fish GHR sequence, and it provides evidence for the conservation of this receptor throughout vertebrate evolution. PMID- 11416051 TI - Estrogen receptor-beta immunoreactivity in luteinizing hormone-releasing hormone neurons of the rat brain. AB - Feedback regulation of luteinizing hormone-releasing hormone (LHRH) neurons by estradiol plays important roles in the neuroendocrine control of reproduction. Recently, we found that the majority of LHRH neurons in the rat contain estrogen receptor-beta (ER-beta) mRNA, whereas, they seemed to lack ER-alpha mRNA expression. In addition, we observed nuclear uptake of (125)I-estrogen by a subset of these cells. These data suggest that ER-beta is the chief receptor isoform mediating direct estrogen effects upon LHRH neurons. To verify the translation of ER-beta protein within LHRH cells, the present studies applied dual-label immunocytochemistry (ICC) to free-floating sections obtained from the preoptic area of rats. The improved ICC method using the silver-gold intensification of nickel-diaminobenzidine chromogen, enabled the observation of nuclear ER-beta-immunoreactivity in the majority of LHRH cells. The incidence of ER-beta expression was similarly high in LHRH neurons of ovariectomized female (87.8 +/- 2.3%, mean +/- SEM), estradiol-primed female (74.9 +/- 3.2%) and intact male (85.0 +/- 4.7%) rats. The presence of ER-beta mRNA, ER-beta immunoreactivity and (125)I-estrogen binding sites in LHRH neurons of the rat provide strong support for the notion that these cells are directly regulated by estradiol, through ER-beta. The gene targets and molecular mechanisms of this regulation remain unknown. PMID- 11416052 TI - Melanin-concentrating hormone (MCH) suppresses thyroid stimulating hormone (TSH) release, in vivo and in vitro, via the hypothalamus and the pituitary. AB - Melanin-concentrating hormone (MCH) is an orexigenic peptide encoded in the pre pro MCH gene. Targeted deletion of MCH causes a phenotype of hypophagia and leanness with an inappropriately high metabolic rate, suggesting a role for MCH in the control of energy balance. In order to further elucidate the mechanism by which MCH controls, energy expenditure, we have investigated the effects of MCH on the hypothalamic pituitary thyroid (HPT) axis. The thyroid axis is important in energy homeostasis and starvation leads to profound suppression of the HPT axis. MCH significantly reduces plasma TSH in vivo at 10 min (0.5 +/- 0.07 ng/ml, p < 0.05, n = 8) and 60 min (0.33 +/- 0.04 ng/ml, p < 0.01, n = 10) compared to saline (0.7 +/- 0.07 ng/ml and 0.69 +/- 0.07 ng/ml respectively) when administered intracerebroventricularly. Release of TRH form hypothalamic explants was significantly reduced in the presence of MCH production (7.1 +/- 0.99 fmol/explant to 2.3 +/- 0.4 fmol/explant p < 0.01, n = 18) and Neuropeptide EI (NEI) (8.47 +/- 1.28 fmol/explant to 4.6 +/- 1.13 p < 0.05, n = 16), a peptide, also encoded in the pre-pro-MCH gene. MCH was also shown to significantly reduce TRH stimulated TSH release from dispersed pituitary cell cultures (basal = 0.5 +/ 0.06 ng/ml, 100 nM TRH = 0.9 +/- 0.2 ng/ml, p < 0.05 0.1 nM MCH = 0.5 +/- 0.1 ng/ml, p < 0.05, 1 nM MCH = 0.3 +/- 0.03 ng/ml, p < 0.01, 10 nM MCH = 0.4 +/- 0.02 ng/ml, p < 0.01, 1000 nM MCH = 0.4 +/- 0.05 ng/ml, P < 0.01, n = 4), although basal release of TSH from these cultures was unaffected. These data suggest a possible role for MCH in the control of energy homeostasis via inhibition of the thyroid axis. PMID- 11416054 TI - A renaissance for measurement error. PMID- 11416055 TI - Hormone replacement therapy (HRT)--risks and benefits. PMID- 11416056 TI - Sick individuals and sick populations. AB - Rose G (Department of Epidemiology, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, UK). Sick individuals and sick populations. International Journal of Epidemiology 1985;14:32--38. Aetiology confronts two distinct issues: the determinants of individual cases, and the determinants of incidence rate. If exposure to a necessary agent is homogeneous within a population, then case/control and cohort methods will fail to detect it: they will only identify markers of susceptibility. The corresponding strategies in control are the 'high-risk' approach, which seeks to protect susceptible individuals, and the population approach, which seeks to control the causes of incidence. The two approaches are not usually in competition, but the prior concern should always be to discover and control the causes of incidence. PMID- 11416058 TI - Commentary: reflections on sick individuals and sick populations. PMID- 11416059 TI - Commentary: causes of incidence and causes of cases--a Durkheimian perspective on Rose. PMID- 11416060 TI - Commentary: a radical future for public health. PMID- 11416061 TI - Commentary: the prevention paradox in lay epidemiology--Rose revisited. PMID- 11416062 TI - War, famine and excess child mortality in Africa: the role of parental education. AB - BACKGROUND: Civilian-targeted warfare and famine constitute two of the greatest public health challenges of our time. Both have devastated many countries in Africa. Social services, and in particular, health services, have been destroyed. Dictatorial and military governments have used the withholding of food as a political weapon to exacerbate human suffering. Under such circumstances, war and famine are expected to have catastrophic impacts on child survival. This study examines the role of parental education in reducing excess child mortality in Africa by considering Tigrai-Ethiopia, which was severely affected by famine and civil war during 1973--1991. METHODS: This study uses data from the 1994 Housing and Population Census of Ethiopia and on communities' vulnerability to food crises. Child mortality levels and trends by various subgroups are estimated using indirect methods of mortality estimation techniques. A Poisson regression model is used to examine the relationship between number of children dead and parental education. RESULTS: Although child mortality is excessively high (about 200 deaths per 1000 births), our results show enormous variations in child mortality by parental education. Child mortality is highest among children born to illiterate mothers and illiterate fathers. Our results also show that the role of parental education in reducing child mortality is great during famine periods. In the communities devastated by war, however, its impact was significant only when the father has above primary education. CONCLUSIONS Our findings suggest that both mother's and father's education are significantly and negatively associated with child mortality, although this effect diminishes over time if the crisis is severe and prolonged. The policy implications of our study include, obviously, reducing armed conflict, addressing food security in a timely manner, and expansion of educational opportunities. PMID- 11416064 TI - Association between vitamin A status and lung function level in children aged 6- 9 years in Wukro wereda, Northern Ethiopia. AB - BACKGROUND: In developing countries, studies using morbidity recalls to evaluate the benefits of vitamin A on respiratory health in children under 6 years of age have been inconclusive. This relationship has not been examined in older children. Spirometric measurements, an objective means of assessing respiratory health, require the subject's collaboration and have been successfully used in children over 6 years of age. This report describes a cross-sectional analysis of the relationship between lung function and vitamin A status in an area endemic to vitamin A deficiency. METHODS: The data on which this report is based were gathered prior to the implementation of a prospective trial of the effect of vitamin A supplementation on lung function level in Northern Ethiopia. Vitamin A status was assessed by the Modified Relative Dose Response (MRDR) method and lung function assessed by spirometry in 702 rural children aged 6--9 years. Demographic, personal health, household, environmental and socioeconomic data were gathered by questionnaire. RESULTS: In children with low vitamin A reserve, the unadjusted forced expiratory volume in one second (FEV(1)) was 48.8 ml (P = 0.006) lower than in those with adequate reserve. This difference was 23.1 ml (P = 0.04) when adjusted for age, gender and height and 14.1 ml (P = 0.20) when adjusted for children's demographic, general health, lung function and household related characteristics. CONCLUSION: Although these findings suggest that vitamin A plays a relatively minor role in determining FEV(1) level, interpretation is limited by the cross-sectional design. Further clarification of its role requires a trial of vitamin A supplementation. PMID- 11416065 TI - Commentary: is vitamin A playing a hidden role in children's lung function? PMID- 11416066 TI - Evidence for a 'healthy pregnant woman effect' in Niakhar, Senegal? AB - BACKGROUND: Although it is generally believed that pregnancy exposes women to a wide variety of excess health risks that go beyond the direct obstetric complications of pregnancy, the epidemiological evidence in support of such excess indirect risks is inconclusive. In this article we attempt to document the contribution of indirect causes of death to maternal mortality in rural Senegal by using an epidemiological approach whereby the time spent during pregnancy and postpartum is considered a transient period of exposure to the health hazards of childbearing. METHODS: We use data from an ongoing demographic surveillance system in Niakhar, Senegal and calculate rate ratios comparing death rates in pregnant or recently pregnant women (exposed) with death rates in other women (unexposed), including and excluding direct obstetric deaths. RESULTS: Between ages 20 and 44, pregnancy does not confer additional risks to women. After excluding direct obstetric deaths, exposed women aged 20--39 have surprisingly lower risks of death than unexposed women of the same age. For the very young (15 19) and the very old (45-49), on the other hand, the excess risks associated with pregnancy are considerable and, among women age 45 or older, persist even after excluding direct obstetric deaths. CONCLUSION: The apparent protective effect of pregnancy on women's health that is observed in this study illustrates the paradoxical nature of the concept of indirect causes of maternal mortality, and the difficulties in measuring the risks of death attributable to the pregnancy. Further studies aimed at separating risks attributable to the pregnancy from those that are incidental to the pregnancy are required. PMID- 11416068 TI - Children's height, health and appetite influence mothers' weaning decisions in rural Senegal. AB - BACKGROUND: In many developing countries, breastfed children have a lower nutritional status than those weaned from 12 months of age. Reverse causality, that is, earlier weaning of healthy and well-nourished children, is a possible explanation. METHODS: Maternal reasons for early and late weaning were investigated in a cohort of 485 rural Senegalese children using structured interviews during two rounds at the ages of 18--28 and 23--33 months, respectively. Length, weight and height were assessed, and dates of weaning were monitored. RESULTS: The mean duration of breastfeeding was 24.1 months (quartiles 21.9 and 26.3). Two-thirds of mothers of breastfed children under 2 stated that they would wean at the age of 2, while for breastfed children aged 2 years, a 'tall and strong' child was the most prevalent criterion. The main reasons for weaning prior to 2 years (N = 244) were that the child ate well from the family plate (60%), that the child was 'tall and strong' (46%) and maternal pregnancy (35%). The main reasons for weaning later than the age of 2 were: a 'little, weak' child (33%), food shortage (25%), illness of the child (24%) and refusal of family food (14%, N = 120). Children breastfed above the age of 2 because they were 'small and weak' had lower mean height-for-age and a greater prevalence of stunting than children breastfed late for other reasons (P < 0.0001). CONCLUSION: The habit of postponing weaning of stunted children very likely explains why breastfed children have lower height-for-age than weaned children in this setting. PMID- 11416069 TI - Commentary: does breastfeeding for longer cause children to be shorter? PMID- 11416070 TI - Clustering of childhood mortality in rural Burkina Faso. AB - BACKGROUND: Childhood mortality is a major public health problem in sub-Saharan Africa. For the implementation of efficient public health systems, knowledge of the spatial distribution of mortality is required. METHODS: Data from a demographic surveillance research project were analysed which comprised information obtained for about 30 000 individuals from 39 villages in northwest Burkina Faso (West Africa) in the period 1993--1998. Total childhood mortality rates were calculated and the geographical distribution of total childhood mortality was investigated. In addition, data from a cohort of 686 children sampled from 16/39 of the villages followed up during a randomized controlled trial in 1999 were also used to validate the results from the surveillance data. A spatial scan statistic was used to test for clusters of total childhood mortality in both space and time. RESULTS: Several statistically significant clusters of higher childhood mortality rates comprising different sets of villages were identified; one specific village was consistently identified in both study populations indicating non-random distribution of childhood mortality. Potential risk factors which were available in the database (ethnicity, religion, distance to nearest health centre) did not explain the spatial pattern. CONCLUSION: The findings indicate non-random clustering of total childhood mortality in the study area. The study may be regarded as a first step in prioritizing areas for follow-up public health efforts. PMID- 11416071 TI - Decreased attendance at routine health activities mediates deterioration in nutritional status of young African children under worsening socioeconomic conditions. AB - BACKGROUND: Economic crisis and sociopolitical instability are generally associated with worsening health and nutrition in developing countries. This study examines the role played by the attendance rate of young children at routine health activities in the deterioration of their nutritional status under adverse social and economic conditions. METHODS: Two nutritional cross-sectional surveys were carried out in two districts of Brazzaville, capital city of The Congo, in 1993 and 1996. They included respectively 2807 and 1695 randomly selected children 4--23 months old. The children's nutritional status was assessed by height-for-age in z-scores. Using embedded general linear regression models, explanatory variables (routine health activities index, socio-demographic context, household economic level, prenatal factors) were tested as potential mediators for the effect of the year of survey on child mean height-for-age. RESULTS: The routine health activities index declined sharply from 1993 to 1996. Its introduction in the regression model including all other explanatory variables led to a sharp decrease in the effect of the year on children's nutritional status, showing the important mediating effect of routine health activities. This result was encountered across all economic categories of households. Other explanatory variables showed more limited mediating effect. CONCLUSIONS: Attendance at preventive health activities should be fostered in African urban communities facing harsh socioeconomic situations to prevent further deterioration in the nutritional status of children. PMID- 11416072 TI - Measuring the local burden of disease. A study of years of life lost in sub Saharan Africa. AB - BACKGROUND: An effective health policy necessitates a reliable characterization of the burden of disease (BOD) by cause. The Global Burden of Disease Study (GBDS) aims to deliver this information. For sub-Saharan Africa (SSA) in particular, the GBDS relies on extrapolations and expert guesses. Its results lack validation by locally measured epidemiological data. METHODS: This study presents locally measured BOD data for a health district in Burkina Faso and compares them to the results of the GBDS for SSA. As BOD indicator, standard years of life lost (age-weighted YLL, discounted with a discount rate of 3%) are used as proposed by the GBDS. To investigate the influence of different age and time preference weights on our results, the BOD pattern is again estimated using, first, YLL with no discounting and no age-weighting, and, second, mortality figures. RESULTS: Our data exhibit the same qualitative BOD pattern as the GBDS results regarding age and gender. We estimated that 53.9% of the BOD is carried by men, whereas the GBDS reported this share to be 53.2%. The ranking of diseases by BOD share, though, differs substantially. Malaria, diarrhoeal diseases and lower respiratory infections occupy the first three ranks in our study and in the GBDS, only differing in their respective order. Protein-energy malnutrition, bacterial meningitis and intestinal nematode infections occupy ranks 5, 6 and 7 in Nouna but ranks 15, 27 and 38 in the GBDS. The results are not sensitive to the different age and time preference weights used. Specifically, the choice of parameters matters less than the choice of indicator. CONCLUSIONS: Local health policy should rather be based on local BOD measurement instead of relying on extrapolations that might not represent the true BOD structure by cause. PMID- 11416073 TI - Effect of misclassification of causes of death in verbal autopsy: can it be adjusted? AB - BACKGROUND: Verbal autopsy (VA) is an indirect method of ascertaining cause of death from information about symptoms and signs obtained from bereaved relatives. This method has been used in several settings to assess cause-specific mortality. However, cause-specific mortality estimates obtained by VA are susceptible to bias due to misclassification of causes of death. One way of overcoming this limitation of VA is to adjust the crude VA estimate of cause-specific mortality fractions (CSMF) using the sensitivity and specificity of the VA tool. This paper explores the application of sensitivity and specificity of VA data obtained from a hospital-based validation study for adjusting the effect of misclassification error in VA data obtained from a demographic surveillance system. METHOD: Data from a multi-centre validation study of 796 adult VA, conducted in Tanzania, Ethiopia and Ghana, were used to explore the effect of distribution of causes of death in the validation study population and the pattern of misclassification on the sensitivity and specificity of VA. VA estimates of CSMF for six causes (acute febrile illness, diarrhoeal diseases, TB/AIDS, cardiovascular disorders, direct maternal causes and injures) were obtained from a demographic surveillance system in Morogoro Rural District in Tanzania. These were adjusted for misclassification error by using sensitivity and specificity values of VA obtained from the validation study in a model proposed for correcting the effect of misclassification error in morbidity prevalence surveys. RESULTS: Sensitivity and specificity of VA differed between the three validation study sites depending on the distribution of causes of death. These differences were explained by variations in the level and pattern of misclassification between sites. When these estimates of sensitivity and specificity were applied to data from the demographic surveillance system with a comparable structure of causes of death the difference between crude and adjusted VA estimates of CSMF ranged from 3 to 83%. CONCLUSION: Estimates of sensitivity and specificity obtained from hospital based validation studies must be used cautiously as a de facto 'gold standard' for adjusting the misclassification error in CSMF derived from VA. It is not possible to use sensitivity and specificity estimates derived from a location specific validation study to adjust for misclassification in VA data from populations with substantially different patterns of cause-specific mortality. PMID- 11416074 TI - A case study of using artificial neural networks for classifying cause of death from verbal autopsy. AB - BACKGROUND: Artificial neural networks (ANN) are gaining prominence as a method of classification in a wide range of disciplines. In this study ANN is applied to data from a verbal autopsy study as a means of classifying cause of death. METHODS: A simulated ANN was trained on a subset of verbal autopsy data, and the performance was tested on the remaining data. The performance of the ANN models were compared to two other classification methods (physician review and logistic regression) which have been tested on the same verbal autopsy data. RESULTS: Artificial neural network models were as accurate as or better than the other techniques in estimating the cause-specific mortality fraction (CSMF). They estimated the CSMF within 10% of true value in 8 out of 16 causes of death. Their sensitivity and specificity compared favourably with that of data-derived algorithms based on logistic regression models. CONCLUSIONS: Cross-validation is crucial in preventing the over-fitting of the ANN models to the training data. Artificial neural network models are a potentially useful technique for classifying causes of death from verbal autopsies. Large training data sets are needed to improve the performance of data-derived algorithms, in particular ANN models. PMID- 11416075 TI - A comparison of cigarette smokers recruited through the Internet or by mail. AB - OBJECTIVES: To compare smokers recruited by mail or through the Internet. METHODS: A questionnaire was mailed to 19,352 inhabitants of Switzerland in 1998, in an effort to enroll them in a smoking cessation trial. The same questionnaire was also available on the Internet. Furthermore, we mailed a survey to a representative sample (n = 1000) of the population of Geneva, Switzerland, in 1996. In this study, we compare three groups: 1027 smokers recruited through the Internet, 2961 volunteer trial participants recruited by mail (response rate 16%), and 211 smokers in the representative sample also recruited by mail (response rate 75%). RESULTS: Smokers self-recruited through the Internet were younger, more educated, more motivated to quit smoking and smoked more cigarettes per day than smokers in the other samples. Compared to trial participants, Internet participants had more negative attitudes towards smoking, higher self efficacy scores, and were more addicted to tobacco. The strength of associations between smoking-related variables was similar in Internet and trial participants. CONCLUSION: As expected, the three groups of smokers differed on several characteristics. However, bias in distributions of variables did not imply bias in associations between variables. Thus, Internet recruitment is a potentially useful method for analytical studies that focus on associations between variables. PMID- 11416076 TI - The methodological quality of randomized controlled trials of homeopathy, herbal medicines and acupuncture. AB - BACKGROUND: To investigate the methodological quality of randomized controlled trials in three areas of complementary medicine. METHODS: The methodological quality of 207 randomized trials collected for five previously published systematic reviews on homeopathy, herbal medicine (Hypericum for depression, Echinacea for common cold), and acupuncture (for asthma and chronic headache) was assessed using a validated scale (the Jadad scale) and single quality items. RESULTS: While the methodological quality of the trials was highly variable, the majority had important shortcomings in reporting and/or methodology. Major problems in most trials were the description of allocation concealment and the reporting of drop-outs and withdrawals. There were relevant differences in single quality components between the different complementary therapies: For example, acupuncture trials reported adequate allocation concealment less often (6% versus 32% of homeopathy and 26% of herb trials), and trials on herbal extracts had better summary scores (mean score 3.12 versus 2.33 for homeopathy and 2.19 for acupuncture trials). Larger trials published more recently in journals listed in Medline and in English language scored significantly higher than trials not meeting these criteria. CONCLUSION: Trials of complementary therapies often have relevant methodological weaknesses. The type of weaknesses varies considerably across interventions. PMID- 11416077 TI - Commentary: science friction--complementary/alternative medicine on the stony road from opinion to evidence. PMID- 11416078 TI - Population mixing and childhood diabetes. AB - BACKGROUND: Exposure to infections, particularly in early life, may modify the risk of developing childhood diabetes. Population mixing, based on the number and diversity of incoming migrants to an area can be used as a proxy measure for exposure to infections. We tested the hypothesis that incidence of childhood Type 1 diabetes is higher in areas of low population mixing. METHODS: Children (<15 years) diagnosed with diabetes between 1986--1994 in Yorkshire, UK (n = 994) were analysed with demographic data and denominator populations from the 1991 UK Census. Population mixing was estimated separately for 'any age' (>1 year) and children (1--15 years) for each area, using the proportion of migrants and an index of diversity based on numbers and origins of migrants. Regression models calculated the effect of 'any age' and childhood population mixing on the incidence of diabetes, controlling for population density, ethnicity and proportion of migrants. RESULTS: Areas with low levels of population mixing of children (bottom decile), were significantly associated with higher incidence of childhood diabetes for 0-14 years (incidence rate ratio [IRR] = 1.46, 95% CI : 1.01--2.11). When stratified by age different effects were observed for childhood population mixing with raised IRR for ages 5-9 (2.23, 95% CI : 1.20--4.11) and 10 14 (1.47, 95% CI : 0.89--2.42), and decreased IRR for 0--4-year-olds (0.56, 95% CI : 0.17--1.82). CONCLUSION: The incidence of childhood diabetes is highest in areas where limited childhood population mixing occurs and the diversity of origins of incoming children is low; those over 4 years are at greatest risk. This is consistent with an infectious hypothesis where absence of stimulation to the developing immune system increases vulnerability to late infectious exposure, which may precipitate diabetes. PMID- 11416080 TI - Cigarette smoking and diabetes mellitus: evidence of a positive association from a large prospective cohort study. AB - OBJECTIVE: Only a few prospective studies have examined the relationship between the frequency of cigarette smoking and the incidence of diabetes mellitus. The purpose of this study was to determine whether greater frequency of cigarette smoking accelerated the development of diabetes mellitus, and whether quitting reversed the effect. METHODS: Data were collected in the Cancer Prevention Study I, a prospective cohort study conducted from 1959 through 1972 by the American Cancer Society where volunteers recruited more than one million acquaintances in 25 US states. From these over one million original participants, 275,190 men and 434,637 women aged > or = 30 years were selected for the primary analysis using predetermined criteria. RESULTS: As smoking increased, the rate of diabetes increased for both men and women. Among those who smoked > or = 2 packs per day at baseline, men had a 45% higher diabetes rate than men who had never smoked; the comparable increase for women was 74%. Quitting smoking reduced the rate of diabetes to that of non-smokers after 5 years in women and after 10 years in men. CONCLUSIONS: A dose-response relationship seems likely between smoking and incidence of diabetes. Smokers who quit may derive substantial benefit from doing so. Confirmation of these observations is needed through additional epidemiological and biological research. PMID- 11416081 TI - Cigarette smoking and glycaemia: the EPIC-Norfolk Study. European Prospective Investigation into Cancer. AB - BACKGROUND: Previous prospective studies have suggested that cigarette smoking may be associated with an increased risk of type 2 diabetes, but the possibility of confounding, particularly by dietary factors has not been fully examined. METHODS: Cross-sectional analysis of the association between cigarette smoking and HbA(1C), a marker of long-term glucose homeostasis in 2704 men and 3385 women, aged 45--74 years who were recruited to a population-based study of diet and chronic disease. RESULTS: Twelve per cent of men and 11% of women reported being current smokers. Mean HbA(1C) was lowest in never smokers, intermediate in former smokers and highest in current smokers. There was a dose-response relationship between HbA(1C) levels and number of cigarettes smoked per day and a positive association with total smoking exposure as measured by pack-years. The unadjusted increase in HbA(1C) for 20 pack-years of smoking was 0.12% (95% CI : 0.09--0.16) in men and 0.12% (95% CI : 0.08--0.17) in women. After adjustment for possible confounders including dietary variables, the values were 0.08% (95% CI : 0.04--0.12) and 0.07% (95% CI : 0.02--0.12) for men and women, respectively. Mean HbA(1C) was inversely related to time since quitting smoking in men. CONCLUSIONS: These results add support to the hypothesis that smoking has long-term effects on glucose homeostasis, an association that cannot be explained by confounding by dietary factors as measured in this study. PMID- 11416082 TI - Commentary: smoking and diabetes--accumulating evidence of a causal link. PMID- 11416083 TI - Prevalence of asthma and other respiratory symptoms in children living near and away from opencast coal mining sites. AB - BACKGROUND: Public concern about respiratory conditions prompted the investigation of asthma and other respiratory diseases in children living near and away from opencast coal mining sites. METHODS: We selected all 4860 children aged 1--11 years from five socioeconomically matched pairs of communities close to (OC) and away from (CC) active opencast sites. A postal questionnaire collected data on health and lifestyle. Outcomes were the cumulative and period prevalence (2 and 12 months) of wheeze, asthma, bronchitis and other respiratory symptoms. RESULTS: The cumulative prevalence of wheeze varied from 30% to 40% across the ten communities, it was 36% in OC and 37% in CC. The cumulative prevalence of asthma was 22% in both OC and CC, varying between 12% and 24%. We found little evidence for associations between living near an opencast site and an increased prevalence of respiratory illnesses, or asthma severity. Some outcomes such as allergies, hayfever, or cough varied little across the study communities. Others, such as the use of asthma medication, the number of severe wheezing attacks in the past year or tonsillitis showed large variation. These similarities and variations were not explained by differences in lifestyle factors or differences in health services delivery and remain unexplained. CONCLUSIONS: There was little evidence of an association between residential proximity to opencast mining sites and cumulative or period prevalence of respiratory illness, or asthma severity. Some variations in health outcomes between communities remained unexplained. PMID- 11416084 TI - Challenges for improving surveillance for pesticide poisoning: policy implications for developing countries. AB - BACKGROUND: Surveillance is a critical public health tool for the control of pesticide poisoning. However, surveillance activities in developing countries are bedevilled by multiple problems, and inferences made from review of flawed data may lead to mistaken policy decisions. METHODS: Results of intensified surveillance from an intervention project in the Western Cape Province of South Africa were compared to the pattern of poisonings reported in routine notifications to the health authorities for a control farming district and in the study district over a 5-year period preceding the study. Intensified surveillance data results were also contrasted with policy approaches based on routine notifications and on Regional Poison Centre reports. RESULTS: Poisoning rates reported in the study area increased almost 10-fold during the intervention period. Compared to intensified surveillance, hospital and health authority sources greatly underestimate the proportion of cases due to occupational poisoning, and overestimate suicide as a proportional cause. In addition, the risks for women appear underestimated from routine notifications. Assumptions that a lack of awareness is responsible for most poisonings are not borne out by the empirical data when reporting is intensified. CONCLUSIONS: Current policy assumptions are faulty, may result in inappropriate blame being attributed to victims and, by relying on information as the main element of education, may shift responsibility onto the individual. Improvements in the surveillance system should aim to restructure the types of data collected, and facilitate intra governmental and inter-sector collaboration. The culture of monitoring based on report writing must change to one of surveillance that leads to intervention. PMID- 11416085 TI - Should programmes for community-level meningococcal vaccination be considered in Australia? An economic evaluation. AB - BACKGROUND: Disease due to serogroup C Neisseria meningitidis is life-threatening and potentially preventable by vaccination. In 1999, the UK instigated mass vaccination after a sustained increase in serogroup C meningococcal disease. In the same year, Victoria, Australia experienced a similar change in disease epidemiology. It is timely to undertake an economic evaluation of options for community vaccination in Australia based on local data. METHODS: Cost effectiveness and cost-benefit analyses of three options for use of polysaccharide vaccine were undertaken for a hypothetical population aged 15--19 years. Baseline analyses assumed 5 years' duration of vaccine protection following a single year of programme implementation. Sensitivity analyses of key variables were performed, including vaccine coverage and effectiveness, case fatality rate and the discount rate. Outcomes included the number of people vaccinated, cases averted, life-years saved and disability-adjusted life-years (DALY) averted. Cost-benefit analysis used lost earnings avoided as a measure of vaccination benefit. RESULTS: Vaccination of people aged 15--19 years in a defined population with a high rate of disease was the most cost-effective option. Compared with no vaccination and assuming 5 years' duration of protection and exclusion of direct cost savings, this resulted in a discounted cost per life year saved of $23,623, a cost per DALY avoided of $21,097 and benefits exceeding costs in discounted terms. The 'break-even' incidence rate for this option with exclusion of direct cost savings was 14.0/100,000. CONCLUSIONS: Community use of polysaccharide vaccination may be cost effective in Australia under certain conditions. Economic evidence favours use of vaccination in well-defined populations with a high rate of disease. Policy decision-making also requires consideration of non-economic factors, including feasibility of implementation and risk perception by the community. PMID- 11416087 TI - Prevalence and association between herpes simplex virus types 1 and 2-specific antibodies in attendees at a sexually transmitted disease clinic. AB - BACKGROUND: Seroprevalence of herpes simplex virus type 1 (HSV-1) and HSV-2 was determined in 1993 and 1998 in a randomly selected study group of 1024 and 654 attendees, respectively, at the sexually transmitted disease (STD) clinic of the University Hospital Rotterdam-Dijkzigt, The Netherlands. Correlations of HSV-1 and HSV-2 seropositivity were investigated. The relationship between HSV-1 and HSV-2 antibodies was also studied. METHODS: Data were collected in a cross sectional study from February 1993 until February 1994 and from January 1998 until December 1998. Glycoprotein G (gG) HSV type specific serum IgG was determined. RESULTS: Seroprevalence of HSV-1 was 68% versus 59% (1993 versus 1998, chi(2)-test P < 0.001), of HSV-2 it was 30% versus 22% (1993 versus 1998, chi(2)-test P < 0.001). Using logistic regression analyses, HSV-1 and HSV-2 seropositivity were significantly associated with age and ethnicity in both groups. In 1993, HSV-1 seropositivity also correlated with lower level of education and female gender, whereas in 1998 it correlated with 'number of sexual partners in the past 6 months' and 'present diagnosis of STD'. In both groups, HSV-2 seropositivity was also more prevalent in females and related to sexual lifestyle variables. In an exposure-disease model, HSV-1 seropositivity was not correlated with HSV-2 seropositivity (odds ratio 1993 = 1.1, 95% CI : 0.8--1.7; odds ratio in 1998 = 1.0, 95% CI : 0.5--1.8). CONCLUSIONS: Seroprevalence of HSV 1 and HSV-2 is falling among STD clinic attendees in Rotterdam. A changing pattern of risk factors for HSV-1 seropositivity indicates increasing sexual transmission of HSV-1. Seropositivity for HSV-2 correlated with known risk factors. A previous HSV-1 infection does not reduce susceptibility to subsequent genital HSV-2 infections. PMID- 11416088 TI - Commentary: developing preventive strategies in Europe. PMID- 11416089 TI - Risk factors for Alzheimer's disease: a population-based, longitudinal study in Manitoba, Canada. AB - BACKGROUND: Current knowledge of risk factors for Alzheimer's disease (AD) is limited. Data from a longitudinal, population-based study of dementia in Manitoba, Canada were used to investigate risk factors for AD. METHODS: Cognitively intact subjects completed a risk factor questionnaire assessing sociodemographic, genetic, environmental, medical and lifestyle exposures. Five years later, 36 subjects had developed AD and 658 remained cognitively intact. RESULTS: Older subjects or those who had fewer years of education were at greater risk of AD. After adjusting for age, education and sex, occupational exposure to fumigants/ defoliants was a significant risk factor for AD (relative risk [RR] = 4.35; 95% CI : 1.05--17.90). A history of migraines increased the risk of AD (RR = 3.49; 95% CI : 1.39--8.77); an even stronger effect was noted among women. Self reported memory loss at baseline was associated with subsequent development of AD (RR = 5.15; 95% CI : 2.36--11.27). Vaccinations and occupational exposure to excessive noise reduced the risk of AD. CONCLUSIONS: Some well-known risk factors for AD were confirmed in this study and potential new risk factors were identified. The association of AD with a history of migraines and occupational exposure to defoliants/fumigants is of particular interest because these are biologically plausible risk factors. PMID- 11416090 TI - Commentary: searching for risks for Alzheimer's disease. PMID- 11416091 TI - Self-rated health and limiting longstanding illness: inter-relationships with morbidity in early adulthood. AB - BACKGROUND: Self-rated health and limiting longstanding illness are both widely used global measures of health, but understanding is poor of their meaning and validity at younger ages. METHODS: We examined the association between self-rated health and limiting longstanding illness and specific health problems at two ages (23 and 33 years), and assessed change over the 10-year period for each health measure relative to another. Longitudinal data were taken from the nationally representative British birth cohort for which health measures were obtained at ages 23 and 33. RESULTS: Self-rated health and limiting longstanding illness were strongly associated with each other as well as with specific health problems, particularly with serious conditions (e.g. epilepsy, cancer, diabetes) and more weakly with less serious conditions (e.g. eczema and hay fever). Rating of overall health and limiting longstanding illness was highly stable during the 10 year period with most, but not all, health change reflecting a deterioration in health status. Deterioration in limiting illness corresponded to an even greater health decline in specific conditions. CONCLUSIONS: Self-rated health and limiting longstanding illness are valid health measures appropriate for use in general health surveys. PMID- 11416092 TI - Japanese and Western diet and risk of idiopathic sudden deafness: a case-control study using pooled controls. AB - BACKGROUND: One of the proposed aetiological mechanisms for idiopathic sudden deafness is vascular disease. However, it is not known whether traditional cardiovascular risk factors, such as particular dietary factors, are associated with this condition. METHODS: A case-control study using pooled controls was conducted in Japan to investigate the relationship between idiopathic sudden deafness and diet. An m:n matched-pairs method was used to obtain age-, gender- and residential district-matched controls from a nationwide database of pooled controls. Food intake was assessed from a self-administered usual food frequency questionnaire that asked about intake of 35 foods (including four drinks). Participants were classified according to the frequency of intake of Western foods and the frequency of intake of traditional Japanese foods. Subgroup analyses were performed using audiometric subtypes of idiopathic sudden deafness. RESULTS: Data were obtained for 164 cases and 20,313 controls. An increased risk of sudden deafness was observed among participants who frequently consumed Western foods (OR = 1.82, 95% CI : 1.14--2.89), and a decreased risk of this condition was observed among participants who frequently consumed Japanese foods (OR = 0.52, 95% CI : 0.33--0.82). A direct association of sudden deafness with Western food intake was evident for flat-type hearing loss. CONCLUSIONS: This study suggests that a largely Western diet might be a risk factor for idiopathic sudden deafness, a traditional Japanese diet might be a preventive factor for this condition, or both. These findings are consistent with the hypothesis that vascular factors are an important cause of idiopathic sudden deafness, although the possibility of residual confounding by unmeasured confounders such as socioeconomic status cannot be ruled out. PMID- 11416093 TI - Impact of smoking habit on medical care use and its costs: a prospective observation of National Health Insurance beneficiaries in Japan. AB - BACKGROUND: To quantify excess medical use associated with smoking, a large prospective cohort study is needed. The authors examined the impact of smoking on medical care use in a large population-based cohort with an accurate data collecting system in Japan. METHOD: The data were derived from a 30-month prospective cohort study of 43,408 National Health Insurance beneficiaries aged 40--79 years living in a rural Japanese community. The smoking habit of beneficiaries was assessed in a baseline survey at the end of 1994. Medical care use and its costs were monitored by linkage with the National Health Insurance claim history files since January 1995. RESULTS: Male smokers incurred 11% more medical costs (after adjustment for age, physical functioning status, alcohol consumption, body mass index and average time spent walking) than 'never smokers' but for female smokers and never smokers the costs were almost the same. This difference was mainly attributable to increased use of inpatient medical care among smokers, especially in males, where per month cost of inpatient care was 33% higher in smokers. Age-group specific analysis in men showed that excess mortality and excess medical cost ratio for smokers peaked in those aged 60--69 years. CONCLUSIONS: Smokers consume excess medical care. Among the population aged 45 years and over, about 4% of total medical costs were attributable to smoking. To pursue both better health and lower medical costs for the nation, a comprehensive programme to reduce tobacco use is needed. PMID- 11416095 TI - Are cardiovascular disease trends driven by gadflies? PMID- 11416096 TI - Social epidemiology: no way back. A response to Zielhuis and Kiemeney. PMID- 11416097 TI - Alcohol consumption and plasma homocysteine: what's brewing? PMID- 11416101 TI - Viruses as therapeutic agents against malignant disease of the central nervous system. PMID- 11416102 TI - Epidermal growth factor receptor signaling and response of cancer cells to ionizing radiation. PMID- 11416103 TI - Cancer fatigue: one drug fails but more are in the pipeline. PMID- 11416104 TI - IARC shifts priorities as research needs change. PMID- 11416105 TI - Studies prompt closer scrutiny of conflict of interest policies. PMID- 11416106 TI - Surgical oncology focusing on minimally invasive surgery, more randomized clinical trials. PMID- 11416109 TI - Obesity, cancer links prompt new recommendations. PMID- 11416111 TI - Reovirus as an oncolytic agent against experimental human malignant gliomas. AB - BACKGROUND: Reovirus is a naturally occurring oncolytic virus that usurps activated Ras-signaling pathways of tumor cells for its replication. Ras pathways are activated in most malignant gliomas via upstream signaling by receptor tyrosine kinases. The purpose of this study was to determine the effectiveness of reovirus as an experimental treatment for malignant gliomas. METHODS: We investigated whether reovirus would infect and lyse human glioma cell lines in vitro. We also tested the effect of injecting live reovirus in vivo on human gliomas grown subcutaneously or orthotopically (i.e., intracerebrally) in mice. Finally, reovirus was tested ex vivo against low-passage cell lines derived from human glioma specimens. All P values were two-sided. RESULTS: Reovirus killed 20 (83%) of 24 established malignant glioma cell lines tested. It caused a dramatic and often complete tumor regression in vivo in two subcutaneous (P =.0002 for both U251N and U87) and in two intracerebral (P =.0004 for U251N and P =.0009 for U87) human malignant glioma mouse models. As expected, serious toxic effects were found in these severely immunocompromised hosts. In a less immunocompromised mouse model, a single intratumoral inoculation of live reovirus led to a dramatic prolongation of survival (compared with control mice treated with dead virus; log rank test, P<.0001 for both U251N and U87 cell lines). The animals treated with live virus also appeared to be healthier and gained body weight (P =.0001). We then tested the ability of reovirus to infect and kill primary cultures of brain tumors removed from patients and found that it killed nine (100%) of nine glioma specimens but none of the cultured meningiomas. CONCLUSIONS: Reovirus has potent activity against human malignant gliomas in vitro, in vivo, and ex vivo. Oncolysis with reovirus may be a potentially useful treatment for a broad range of human cancers. PMID- 11416112 TI - Randomized adjuvant chemotherapy trial in high-risk, lymph node-negative breast cancer patients identified by urokinase-type plasminogen activator and plasminogen activator inhibitor type 1. AB - BACKGROUND: Most patients with lymph node-negative breast cancer are cured by locoregional treatment; however, about 30% relapse. Because traditional histomorphologic and clinical factors fail to identify the high-risk patients who may benefit from adjuvant chemotherapy, other prognostic factors are needed. In a unicenter study, we have found that levels of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor type 1 (PAI-1) in the primary tumor are predictive of disease recurrence. Thus, we designed the Chemo N(0) prospective randomized multicenter therapy trial to investigate further whether uPA and PAI-1 are such prognostic factors and whether high-risk patients identified by these factors benefit from adjuvant chemotherapy. After 4.5 years, we present results of the first interim analysis. METHODS: We studied 556 patients with lymph node-negative breast cancer. The median follow-up was 32 months. All patients with low tumor levels of uPA (< or = 3 ng/mg of protein) and of PAI-1 (< or = 14 ng/mg of protein) were observed. Patients with high tumor levels of uPA (> 3 ng/mg of protein) and/or of PAI-1 (> 14 ng/mg of protein) were randomly assigned to combination chemotherapy or subjected to observation only. All statistical tests were two-sided. RESULTS: A total of 241 patients had low levels of uPA and PAI-1, and 315 had elevated levels of uPA and/or PAI-1. The estimated 3-year recurrence rate for patients with low tumor levels of uPA and PAI-1 (low-risk group) was 6.7% (95% confidence interval [CI] = 2.5% to 10.8%). This rate for patients with high tumor levels of uPA and/or PAI-1 (high-risk group) was 14.7% (95% CI = 8.5% to 20.9%) (P = 0.006). First interim analysis suggests that high-risk patients in the chemotherapy group benefit, with a 43.8% lower estimated probability of disease recurrence at 3 years than high-risk patients in the observation group (intention-to-treat analysis: relative risk = 0.56; 95% CI = 0.25 to 1.28), but further follow-up is needed for confirmation. CONCLUSIONS: Using uPA and PAI-1, we have been able to classify about half of the patients with lymph node-negative breast cancer as low risk, for whom adjuvant chemotherapy may be avoided, and half as high risk, who appear to benefit from adjuvant chemotherapy. PMID- 11416113 TI - Epidermal growth factor receptor as a genetic therapy target for carcinoma cell radiosensitization. AB - BACKGROUND: Exposure of human cancer cells to ionizing radiation activates the epidermal growth factor receptor (EGFR), which, in turn, mediates a cytoprotective response that reduces the cells' sensitivity to ionizing radiation. Overexpression of a dominant-negative EGFR mutant, EGFR-CD533, disrupts the cytoprotective response by preventing radiation-induced activation of the receptor and its downstream effectors. To investigate whether gene therapy with EGFR-CD533 has the potential to increase tumor cell radiosensitivity, we introduced an adenoviral vector containing EGFR-CD533 into xenograft tumors in nude mice and evaluated the tumor response to ionizing radiation. METHODS: Xenograft tumors established from the human mammary carcinoma cell line MDA-MB 231 were transduced via infusion with the adenoviral vector Ad-EGFR-CD533 or a control vector containing the beta-galactosidase gene, Ad-LacZ. The transduced tumors were then exposed to radiation in the therapeutic dose range, and radiation-induced EGFR activation was assessed by examining the tyrosine phosphorylation of immunoprecipitated EGFR. Radiosensitization was determined in vitro by colony-formation assays. All statistical tests were two-sided. RESULTS: The transduction efficiency of MDA-MB-231 tumors by Ad-LacZ was 44%. Expression of EGFR-CD533 in tumors reduced radiation-induced EGFR activation by 2.94-fold (95% confidence interval [CI] = 2.23 to 4.14). The radiosensitivity of Ad-EGFR CD533-transduced tumors was statistically significantly higher (46%; P<.001) than that of Ad-LacZ-transduced tumors, yielding a dose-enhancement ratio of 1.85 (95% CI = 1.54 to 2.51). CONCLUSIONS: Transduction of MDA-MB-231 xenograft tumors with Ad-EGFR-CD533 conferred a dominant-negative EGFR phenotype and induced tumor radiosensitization. Therefore, disruption of EGFR function through overexpression of EGFR-CD533 may hold promise as a gene therapeutic approach to enhance the sensitivity of tumor cells to ionizing radiation. PMID- 11416114 TI - Bone mass and breast cancer risk in older women: differences by stage at diagnosis. AB - BACKGROUND: Older women with low bone mineral density (BMD) have a decreased incidence of breast cancer. It is not known whether this association is confined to early-stage, slow-growing tumors. METHODS: We prospectively studied 8905 women who were 65 years of age or older during the period from 1986 through 1988 and had no history of breast cancer. At study entry, we used single-photon absorptiometry to measure each woman's BMD at three skeletal sites: the wrist, forearm, and heel. The women were followed for a mean of 6.5 years for the occurrence of breast cancer. All statistical tests were two-sided. RESULTS: During 57 516 person-years of follow-up, 315 women developed primary invasive or in situ breast cancer. Multivariate analyses that adjusted for age, obesity, and other covariates revealed that the risk of breast cancer for women in the highest quartile of BMD for all three skeletal sites was 2.7 (95% confidence interval [CI] = 1.4 to 5.3) times greater than that for women in the lowest quartile at all three skeletal sites. The magnitude of increased risk associated with high BMD differed by the stage of disease at diagnosis and was greater for more advanced tumors (relative risk [RR] for TNM [i.e., tumor-lymph node-metastasis] stage II or higher tumors = 5.6; 95% CI = 1.2 to 27.4) than for early-stage disease (RR for in situ/TNM stage I tumors = 2.2; 95% CI = 1.0 to 4.8). CONCLUSIONS: Elderly women with high BMD have an increased risk of breast cancer, especially advanced cancer, compared with women with low BMD. These findings suggest an association between osteoporosis and invasive breast cancer, two of the most prevalent conditions affecting an older woman's health. PMID- 11416115 TI - Helicobacter pylori seropositivity as a risk factor for pancreatic cancer. AB - BACKGROUND: Pancreatic cancer is among the most fatal cancers worldwide and one for which few preventable risk factors have been established. Gastric carriage of Helicobacter pylori, particularly cytotoxin-associated gene-A-positive (CagA+) strains, is known to be a risk factor for peptic ulcer disease and gastric cancer and may have a similar etiologic relationship with pancreatic cancer. METHODS: We investigated the association of H. pylori carriage and exocrine pancreatic cancer in a nested case-control study within the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study cohort of 29 133 male Finnish smokers aged 50-69 years at baseline. Case subjects (n = 121) were matched on date of baseline serum collection, study center, age, trial intervention, and completion of the dietary questionnaire to 226 control subjects who were alive at the time the matching case subject was diagnosed and who remained free of cancer, during up to 10 years of follow-up. Levels of immunoglobulin G antibodies to H. pylori whole-cell and CagA+ antigens from stored baseline serum were measured by enzyme-linked immunosorbent assay. Smoking-adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were estimated by use of conditional logistic regression. Statistical tests were two-sided. RESULTS: Seroprevalence of H. pylori was 82% and 73% among case and control subjects, respectively. Compared with seronegative subjects, those with H. pylori or CagA+ strains were at statistically significantly elevated risk of pancreatic cancer (OR = 1.87 [95% CI = 1.05 to 3.34]; OR = 2.01 [95% CI = 1.09 to 3.70], respectively). CONCLUSIONS: Our findings support a possible role for H. pylori carriage in the development of exocrine pancreatic cancer. PMID- 11416116 TI - Prospective study of recreational physical activity and ovarian cancer. AB - BACKGROUND: It has been hypothesized that physical activity may reduce the risk of ovarian cancer by decreasing estrogen levels, reducing body fat, and reducing the frequency of ovulation. Epidemiologic studies of this relationship have obtained inconsistent results. The only prospective study to date reported a positive association between frequent vigorous exercise and ovarian cancer risk. We further evaluated this relationship in the Nurses' Health Study cohort. METHODS: Participation in recreational physical activity was assessed by questionnaire in 1980, 1982, 1986, 1988, 1992, and 1994, with questions assessing exercise frequency, duration, and intensity. Results were adjusted for age, parity, oral contraceptive use, tubal ligation, and other risk factors for ovarian cancer. All statistical tests were two-sided. RESULTS: During a 16-year follow-up (from 1980 to 1996), 1.2 million person-years were accrued by 92 825 cohort members, and 377 cases of epithelial ovarian cancer were confirmed. The relative risk (RR) of ovarian cancer for women engaging in recreational physical activity for 7 hours or more per week compared with those reporting less than 1 hour per week was 0.80 (95% confidence interval [CI] = 0.49 to 1.32; P(trend) =.59). When both the frequency and intensity of activity were taken into account, activity level was also not associated with a reduced risk of ovarian cancer. Compared with inactive women, participants reporting high activity in terms of metabolic equivalent task hours (MET hours) were at greater risk of ovarian cancer (RR for 20 to <30 MET hours/week = 1.84 [95% CI = 1.12 to 3.02]; RR for >30 MET hours/week = 1.27 [95% CI = 0.75 to 2.14]). CONCLUSIONS: Overall, results did not suggest an inverse association between recreational physical activity and ovarian cancer. The possibility of a modest increase in risk with frequent vigorous activity requires further investigation. PMID- 11416117 TI - Re: Stem cell transplant numbers decline; research continues. PMID- 11416118 TI - Re: Comparison of three management strategies for patients with atypical squamous cells of undetermined significance: baseline results from a randomized trial. PMID- 11416120 TI - Re: Comparison of three management strategies for patients with atypical squamous cells of undetermined significance: baseline results from a randomized trial. PMID- 11416122 TI - Re: Cellular telephones and cancer--a nationwide cohort study in Denmark. PMID- 11416123 TI - Promoter-specific shifts in transcription initiation conferred by yeast TFIIB mutations are determined by the sequence in the immediate vicinity of the start sites. AB - The general transcription factor IIB (TFIIB) is required for transcription of class II genes by RNA polymerase II. Previous studies demonstrated that mutations in the Saccharomyces cerevisiae SUA7 gene, which encodes TFIIB, can alter transcription initiation patterns in vivo. To further delineate the functional domain and residues of TFIIB involved in transcription start site utilization, a genetic selection was used to isolate S. cerevisiae TFIIB mutants exhibiting downstream shifts in transcription initiation in vivo. Both dominant and recessive mutations conferring downstream shifts were identified at multiple positions within a highly conserved homology block in the N-terminal region of the protein. The TFIIB mutations conferred downstream shifts in transcription initiation at the ADH1 and CYC1 promoters, whereas no significant shifts were observed at the HIS3 promoter. Analysis of a series of ADH1-HIS3 hybrid promoters and variant ADH1 and HIS3 promoters containing insertions, deletions, or site directed base substitutions revealed that the feature that renders a promoter sensitive to TFIIB mutations is the sequence in the immediate vicinity of the normal start sites. We discuss these results in light of possible models for the mechanism of start site utilization by S. cerevisiae RNA polymerase II and the role played by TFIIB. PMID- 11416124 TI - Phosphorylation of MafA is essential for its transcriptional and biological properties. AB - We previously described the identification of quail MafA, a novel transcription factor of the Maf bZIP (basic region leucine zipper) family, expressed in the differentiating neuroretina (NR). In the present study, we provide the first evidence that MafA is phosphorylated and that its biological properties strongly rely upon phosphorylation of serines 14 and 65, two residues located in the transcriptional activating domain within a consensus for phosphorylation by mitogen-activated protein kinases and which are conserved among Maf proteins. These residues are phosphorylated by ERK2 but not by p38, JNK, and ERK5 in vitro. However, the contribution of the MEK/ERK pathway to MafA phosphorylation in vivo appears to be moderate, implicating another kinase. The integrity of serine 14 and serine 65 residues is required for transcriptional activity, since their mutation into alanine severely impairs MafA capacity to activate transcription. Furthermore, we show that the MafA S14A/S65A mutant displays reduced capacity to induce expression of QR1, an NR-specific target of Maf proteins. Likewise, the integrity of serines 14 and 65 is essential for the MafA ability to stimulate expression of crystallin genes in NR cells and to induce NR-to-lens transdifferentiation. Thus, the MafA capacity to induce differentiation programs is dependent on its phosphorylation. PMID- 11416125 TI - Characterization of the Crithidia fasciculata mRNA cycling sequence binding proteins. AB - The Crithidia fasciculata cycling sequence binding protein (CSBP) binds with high specificity to sequence elements in several mRNAs that accumulate periodically during the cell cycle. Mutations in these sequence elements abolish both cycling of the mRNA and binding of CSBP. Two genes, CSBPA and CSBPB, encoding putative subunits of CSBP have been cloned and were found to be present in tandem on the same DNA molecule and to be closely related. CSBPA and CSBPB are predicted to encode proteins with sizes of 35.6 and 42.0 kDa, respectively. Both CSBPA and CSBPB proteins have a predicted coiled-coil domain near the N terminus and a novel histidine and cysteine motif near the C terminus. The latter motif is conserved in other trypanosomatid species. Gel sieving chromatography and glycerol gradient sedimentation results indicate that CSBP has a molecular mass in excess of 200 kDa and an extended structure. Recombinant CSBPA and CSBPB also bind specifically to the cycling sequence and together can be reconstituted to give an RNA gel shift similar to that of purified CSBP. Proteins in cell extracts bind to an RNA probe containing six copies of the cycling sequence. The RNA protein complexes contain both CSBPA and CSBPB, and the binding activity cycles in near synchrony with target mRNA levels. CSBPA and CSBPB mRNA and protein levels show little variation throughout the cell cycle, suggesting that additional factors are involved in the cyclic binding to the cycling sequence elements. PMID- 11416126 TI - Dual roles of RNA helicase A in CREB-dependent transcription. AB - RNA helicase A (RHA) is a member of an ATPase/DNA and RNA helicase family and is a homologue of Drosophila maleless protein (MLE), which regulates X-linked gene expression. RHA is also a component of holo-RNA polymerase II (Pol II) complexes and recruits Pol II to the CREB binding protein (CBP). The ATPase and/or helicase activity of RHA is required for CREB-dependent transcription. To further understand the role of RHA on gene expression, we have identified a 50-amino-acid transactivation domain that interacts with Pol II and termed it the minimal transactivation domain (MTAD). The protein sequence of this region contains six hydrophobic residues and is unique to RHA homologues and well conserved. A mutant with this region deleted from full-length RHA decreased transcriptional activity in CREB-dependent transcription. In addition, mutational analyses revealed that several tryptophan residues in MTAD are important for the interaction with Pol II and transactivation. These mutants had ATP binding and ATPase activities comparable to those of wild-type RHA. A mutant lacking ATP binding activity was still able to interact with Pol II. In CREB-dependent transcription, the transcriptional activity of each of these mutants was less than that of wild-type RHA. The activity of the double mutant lacking both functions was significantly lower than that of each mutant alone, and the double mutant had a dominant negative effect. These results suggest that RHA could independently regulate CREB dependent transcription either through recruitment of Pol II or by ATP-dependent mechanisms. PMID- 11416127 TI - PLIP, a novel splice variant of Tip60, interacts with group IV cytosolic phospholipase A(2), induces apoptosis, and potentiates prostaglandin production. AB - The group IV cytosolic phospholipase A(2) (cPLA(2)) has been localized to the nucleus (M. R. Sierra-Honigmann, J. R. Bradley, and J. S. Pober, Lab. Investig. 74:684-695, 1996) and is known to translocate from the cytosolic compartment to the nuclear membrane (S. Glover, M. S. de Carvalho, T. Bayburt, M. Jonas, E. Chi, C. C. Leslie, and M. H. Gelb, J. Biol. Chem. 270:15359-15367, 1995; A. R. Schievella, M. K. Regier, W. L. Smith, and L. L. Lin, J. Biol. Chem. 270:30749 30754, 1995). We hypothesized that nuclear proteins interact with cPLA(2) and participate in the functional effects of this translocation. We have identified a nuclear protein, cPLA(2)-interacting protein (PLIP), a splice variant of human Tip60, which interacts with the amino terminal region of cPLA(2). Like Tip60, PLIP cDNA includes the MYST domain containing a C2HC zinc finger and well conserved similarities to acetyltransferases. Both PLIP and Tip60 coimmunoprecipitate and colocalize with cPLA(2) within the nuclei of transfected COS cells. A polyclonal antibody raised to PLIP recognizes both PLIP and Tip60. Endogenous Tip60 and/or PLIP in rat mesangial cells is localized to the nucleus in response to serum deprivation. Nuclear localization coincides temporally with apoptosis. PLIP expression, mediated by adenoviral gene transfer, potentiates serum deprivation-induced prostaglandin E(2) (PGE(2)) production and apoptosis in mouse mesangial cells from cPLA(2)(+/+) mice but not in mesangial cells derived from cPLA(2)(-/-) mice. Thus PLIP, a splice variant of Tip60, interacts with cPLA(2) and potentiates cPLA(2)-mediated PGE(2) production and apoptosis. PMID- 11416128 TI - Deubiquitination step in the endocytic pathway of yeast plasma membrane proteins: crucial role of Doa4p ubiquitin isopeptidase. AB - The Fur4p uracil permease, like most yeast plasma membrane proteins, undergoes ubiquitin-dependent endocytosis and is then targeted to the vacuole (equivalent to the mammalian lysosome) for degradation. The cell surface ubiquitination of Fur4p is mediated by the essential Rsp5p ubiquitin ligase. Ubiquitination of Fur4p occurs on two target lysines, which receive two ubiquitin moieties linked through ubiquitin Lys63, a type of linkage (termed UbK63) different from that involved in proteasome recognition. We report that pep4 cells deficient for vacuolar protease activities accumulate vacuolar unubiquitinated Fur4p. In contrast, pep4 cells lacking the Doa4p ubiquitin isopeptidase accumulate ubiquitin-conjugated Fur4p. These data suggest that Fur4p undergoes Doa4p dependent deubiquitination prior to vacuolar degradation. Compared to pep4 cells, pep4 doa4 cells have huge amounts of membrane-bound ubiquitin conjugates. This indicates that Doa4p plays a general role in the deubiquitination of membrane bound proteins, as suggested by reports describing the suppression of some doa4 phenotypes in endocytosis and vacuolar protein sorting mutants. Some of the small ubiquitin-linked peptides that are a hallmark of Doa4 deficiency are not present in rsp5 mutant cells or after overproduction of a variant ubiquitin modified at Lys 63 (UbK63R). These data suggest that the corresponding peptides are degradation products of Rsp5p substrates and probably of ubiquitin conjugates carrying UbK63 linkages. Doa4p thus appears to be involved in the deubiquitination of endocytosed plasma membrane proteins, some of them carrying UbK63 linkages. PMID- 11416129 TI - Schizosaccharomyces pombe cells lacking the amino-terminal catalytic domains of DNA polymerase epsilon are viable but require the DNA damage checkpoint control. AB - In Schizosaccharomyces pombe, the catalytic subunit of DNA polymerase epsilon (Pol epsilon) is encoded by cdc20(+) and is essential for chromosomal DNA replication. Here we demonstrate that the N-terminal half of Pol epsilon that includes the highly conserved polymerase and exonuclease domains is dispensable for cell viability, similar to observations made with regard to Saccharomyces cerevisiae. However, unlike budding yeast, we find that fission yeast cells lacking the N terminus of Pol epsilon (cdc20(DeltaN-term)) are hypersensitive to DNA-damaging agents and have a cell cycle delay. Moreover, the viability of cdc20(DeltaN-term) cells is dependent on expression of rad3(+), hus1(+), and chk1(+), three genes essential for the DNA damage checkpoint control. These data suggest that in the absence of the N terminus of Pol epsilon, cells accumulate DNA damage that must be repaired prior to mitosis. Our observation that S phase occurs more slowly for cdc20(DeltaN-term) cells suggests that DNA damage might result from defects in DNA synthesis. We hypothesize that the C-terminal half of Pol epsilon is required for assembly of the replicative complex at the onset of S phase. This unique and essential function of the C terminus is preserved in the absence of the N-terminal catalytic domains, suggesting that the C terminus can interact with and recruit other DNA polymerases to the site of initiation. PMID- 11416130 TI - The Drosophila homolog of mammalian zinc finger factor MTF-1 activates transcription in response to heavy metals. AB - Metallothioneins (MTs) are short, cysteine-rich proteins for heavy metal homeostasis and detoxification; they bind a variety of heavy metals and also act as radical scavengers. Transcription of mammalian MT genes is activated by heavy metal load via the metal-responsive transcription factor 1 (MTF-1), an essential zinc finger protein whose elimination in mice leads to embryonic lethality due to liver decay. Here we characterize the Drosophila homolog of vertebrate MTF-1 (dMTF-1), a 791-amino-acid protein which is most similar to its mammalian counterpart in the DNA-binding zinc finger region. Like mammalian MTF-1, dMTF-1 binds to conserved metal-responsive promoter elements (MREs) and requires zinc for DNA binding, yet some aspects of heavy metal regulation have also been subject to divergent evolution between Drosophila and mammals. dMTF-1, unlike mammalian MTF-1, is resistant to low pH (6 to 6.5). Furthermore, mammalian MT genes are activated best by zinc and cadmium, whereas in Drosophila cells, cadmium and copper are more potent inducers than zinc. The latter species difference is most likely due to aspects of heavy metal metabolism other than MTF 1, since in transfected mammalian cells, dMTF-1 responds to zinc like mammalian MTF-1. Heavy metal induction of both Drosophila MTs is abolished by double stranded RNA interference: small amounts of cotransfected double-stranded RNA of dMTF-1 but not of unrelated control RNA inhibit the response to both the endogenous dMTF-1 and transfected dMTF-1. These data underline an important role for dMTF-1 in MT gene regulation and thus heavy metal homeostasis. PMID- 11416131 TI - Novel CD28-responsive enhancer activated by CREB/ATF and AP-1 families in the human interleukin-2 receptor alpha-chain locus. AB - The interaction of interleukin-2 (IL-2) with its receptor (IL-2R) critically regulates the T-cell immune response, and the alpha chain CD25/IL-2Ralpha is required for the formation of the high-affinity receptor. Tissue-specific, inducible expression of the IL-2Ralpha gene is regulated by at least three positive regulatory regions (PRRI, PRRII, and PRRIII), but none responded to CD28 engagement in gene reporter assays although CD28 costimulation strongly amplifies IL-2Ralpha gene transcription. By DNase I hypersensitivity analysis, we have identified a novel TCR-CD3- and CD28-responsive enhancer (CD28rE) located 8.5 kb 5' of the IL-2Ralpha gene. PRRIV/CD28rE contains a functional CRE/TRE element required for CD28 signaling. The T-cell-specific, CD28-responsive expression of the IL-2Ralpha gene appears controlled through PRRIV/CD28rE by cooperation of CREB/ATF and AP-1 family transcription factors. PMID- 11416132 TI - Site-specific recognition of a 70-base-pair element containing d(GA)(n) repeats mediates bithoraxoid polycomb group response element-dependent silencing. AB - Polycomb group proteins act through Polycomb group response elements (PREs) to maintain silencing at homeotic loci. The minimal 1.5-kb bithoraxoid (bxd) PRE contains a region required for pairing-sensitive repression and flanking regions required for maintenance of embryonic silencing. Little is known about the identity of specific sequences necessary for function of the flanking regions. Using gel mobility shift analysis, we identify DNA binding activities that interact specifically with a multipartite 70-bp fragment (MHS-70) downstream of the pairing-sensitive sequence. Deletion of MHS-70 in the context of a 5.1-kb bxd Polycomb group response element derepresses maintenance of silencing in embryos. A partially purified binding activity requires multiple, nonoverlapping d(GA)(3) repeats for MHS-70 binding in vitro. Mutation of d(GA)(3) repeats within MHS-70 in the context of the 5.1-kb bxd PRE destabilizes maintenance of silencing in a subset of cells in vivo but gives weaker derepression than deletion of MHS-70. These results suggest that d(GA)(3) repeats are important for silencing but that other sequences within MHS-70 also contribute to silencing. Antibody supershift assays and Western analyses show that distinct isoforms of Polyhomeotic and two proteins that recognize d(GA)(3) repeats, the TRL/GAGA factor and Pipsqueak (Psq), are present in the MHS-70 binding activity. Mutations in Trl and psq enhance homeotic phenotypes of ph, indicating that TRL/GAGA factor and Psq are enhancers of Polycomb which have sequence-specific DNA binding activity. These studies demonstrate that site-specific recognition of the bxd PRE by d(GA)(n) repeat binding activities mediates PcG-dependent silencing. PMID- 11416133 TI - Transactivation by the p65 subunit of NF-kappaB in response to interleukin-1 (IL 1) involves MyD88, IL-1 receptor-associated kinase 1, TRAF-6, and Rac1. AB - We have examined the involvement of components of the interleukin-1 (IL-1) signaling pathway in the transactivation of gene expression by the p65 subunit of NF-kappaB. Transient transfection of cells with plasmids encoding wild-type MyD88, IL-1 receptor-associated kinase 1 (IRAK-1), and TRAF-6 drove p65-mediated transactivation. In addition, dominant negative forms of MyD88, IRAK-1, and TRAF 6 inhibited the IL-1-induced response. In cells lacking MyD88 or IRAK-1, no effect of IL-1 was observed. Together, these results indicate that MyD88, IRAK-1, and TRAF-6 are important downstream regulators of IL-1-mediated p65 transactivation. We have previously shown that the low-molecular-weight G protein Rac1 is involved in this response. Constitutively active RacV12-mediated transactivation was not inhibited by dominant negative MyD88, while dominant negative RacN17 inhibited the MyD88-driven response, placing Rac1 downstream of MyD88 on this pathway. Dominant negative RacN17 inhibited wild-type IRAK-1- and TRAF-6-induced transactivation, and in turn, dominant negative IRAK-1 and TRAF-6 inhibited the RacV12-driven response, suggesting a mutual codependence of Rac1, IRAK-1, and TRAF-6 in regulating this pathway. Finally, Rac1 was found to associate with the receptor complex via interactions with both MyD88 and the IL-1 receptor accessory protein. A pathway emanating from MyD88 and involving IRAK-1, TRAF-6, and Rac1 is therefore involved in transactivation of gene expression by the p65 subunit of NF-kappaB in response to IL-1. PMID- 11416134 TI - Redistribution of glycolipid raft domain components induces insulin-mimetic signaling in rat adipocytes. AB - Caveolae and caveolin-containing detergent-insoluble glycolipid-enriched rafts (DIG) have been implicated to function as plasma membrane microcompartments or domains for the preassembly of signaling complexes, keeping them in the basal inactive state. So far, only limited in vivo evidence is available for the regulation of the interaction between caveolae-DIG and signaling components in response to extracellular stimuli. Here, we demonstrate that in isolated rat adipocytes, synthetic intracellular caveolin binding domain (CBD) peptide derived from caveolin-associated pp59(Lyn) (10 to 100 microM) or exogenous phosphoinositolglycan derived from glycosyl-phosphatidylinositol (GPI) membrane protein anchor (PIG; 1 to 10 microM) triggers the concentration-dependent release of caveolar components and the GPI-anchored protein Gce1, as well as the nonreceptor tyrosine kinases pp59(Lyn) and pp125(Fak), from interaction with caveolin (up to 45 to 85%). This dissociation, which parallels redistribution of the components from DIG to non-DIG areas of the adipocyte plasma membrane (up to 30 to 75%), is accompanied by tyrosine phosphorylation and activation of pp59(Lyn) and pp125(Fak) (up to 8- and 11-fold) but not of the insulin receptor. This correlates well to increased tyrosine phosphorylation of caveolin and the insulin receptor substrate protein 1 (up to 6- and 15-fold), as well as elevated phosphatidylinositol-3' kinase activity and glucose transport (to up to 7- and 13 fold). Insulin-mimetic signaling by both CBD peptide and PIG as well as redistribution induced by CBD peptide, but not by PIG, was blocked by synthetic intracellular caveolin scaffolding domain (CSD) peptide. These data suggest that in adipocytes a subset of signaling components is concentrated at caveolae-DIG via the interaction between their CBD and the CSD of caveolin. These inhibitory interactions are relieved by PIG. Thus, caveolae-DIG may operate as signalosomes for insulin-independent positive cross talk to metabolic insulin signaling downstream of the insulin receptor based on redistribution and accompanying activation of nonreceptor tyrosine kinases. PMID- 11416135 TI - GCN5 dependence of chromatin remodeling and transcriptional activation by the GAL4 and VP16 activation domains in budding yeast. AB - Chromatin-modifying enzymes such as the histone acetyltransferase GCN5 can contribute to transcriptional activation at steps subsequent to the initial binding of transcriptional activators. However, few studies have directly examined dependence of chromatin remodeling in vivo on GCN5 or other acetyltransferases, and none have examined remodeling via nucleosomal activator binding sites. In this study, we have monitored chromatin perturbation via nucleosomal binding sites in the yeast episome TALS by GAL4 derivatives in GCN5(+) and gcn5Delta yeast cells. The strong activator GAL4 shows no dependence on GCN5 for remodeling TALS chromatin, whereas GAL4-estrogen receptor-VP16 shows substantial, albeit not complete, GCN5 dependence. Mini-GAL4 derivatives having weakened interactions with TATA-binding protein and TFIIB exhibit a strong dependence on GCN5 for both transcriptional activation and TALS remodeling not seen for native GAL4. These results indicate that GCN5 can contribute to chromatin remodeling at activator binding sites and that dependence on coactivator function for a given activator can vary according to the type and strength of contacts that it makes with other factors. We also found a weaker dependence for chromatin remodeling on SPT7 than on GCN5, indicating that GCN5 can function via pathways independent of the SAGA complex. Finally, we examine dependence on GCN5 and SWI-SNF at two model promoters and find that although these two chromatin-remodeling and/or modification activities may sometimes work together, in other instances they act in complementary fashion. PMID- 11416136 TI - The EphA8 receptor regulates integrin activity through p110gamma phosphatidylinositol-3 kinase in a tyrosine kinase activity-independent manner. AB - Recent genetic studies suggest that ephrins may function in a kinase-independent Eph receptor pathway. Here we report that expression of EphA8 in either NIH 3T3 or HEK293 cells enhanced cell adhesion to fibronectin via alpha(5)beta(1)- or beta(3) integrins. Interestingly, a kinase-inactive EphA8 mutant also markedly promoted cell attachment to fibronectin in these cell lines. Using a panel of EphA8 point mutants, we have demonstrated that EphA8 kinase activity does not correlate with its ability to promote cell attachment to fibronectin. Analysis using EphA8 extracellular and intracellular domain mutants has revealed that enhanced cell adhesion is dependent on ephrin A binding to the extracellular domain and the juxtamembrane segment of the cytoplasmic domain of the receptor. EphA8-promoted adhesion was efficiently inhibited by wortmannin, a phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor. Additionally, we found that EphA8 had associated PI 3-kinase activity and that the p110gamma isoform of PI 3-kinase is associated with EphA8. In vitro binding experiments revealed that the EphA8 juxtamembrane segment was sufficient for the formation of a stable complex with p110gamma. Similar results were obtained in assay using cells stripped of endogenous ephrin A ligands by treatment with preclustered ephrin A5 Fc proteins. In addition, a membrane-targeted lipid kinase-inactive p110gamma mutant was demonstrated to stably associate with EphA8 and suppress EphA8 promoted cell adhesion to fibronectin. Taken together, these results suggest the presence of a novel mechanism by which the EphA8 receptor localizes p110gamma PI 3-kinase to the plasma membrane in a tyrosine kinase-independent fashion, thereby allowing access to lipid substrates to enable the signals required for integrin mediated cell adhesion. PMID- 11416137 TI - Requirement of CDC45 for postimplantation mouse development. AB - CDC45 is required for the initiation of DNA replication in Saccharomyces cerevisiae and functions as a DNA polymerase alpha loading factor in Xenopus, but its role in mammalian DNA replication is unknown. To investigate the genetic and physiological functions of CDC45, we used a gene targeting strategy to generate mice lacking a functional CDC45 gene. Homozygous mutant mice lacking a functional CDC45 gene underwent uterine implantation and induced uterine decidualization but did not develop substantially thereafter. Detailed analysis of CDC45 null embryos cultured in vitro revealed impaired proliferation of the inner cell mass. These findings make CDC45 the only putative replication factor experimentally proven to be essential for mammalian development. The CDC45 gene localizes to human chromosome 22q11.2 in the DiGeorge syndrome critical region (DGCR). Almost 90% of individuals with congenital cardiac and craniofacial defects have a monoallelic deletion in the DGCR that includes CDC45. We report here that heterozygous mutant mice develop into adulthood without any apparent abnormalities, so that it is unlikely that hemizygosity of CDC45 alone is responsible for the cardiac and craniofacial defects in the congenital syndromes. PMID- 11416138 TI - Characterization of mediator complexes from HeLa cell nuclear extract. AB - A number of mammalian multiprotein complexes containing homologs of Saccharomyces cerevisiae Mediator subunits have been described recently. High-molecular-mass complexes (1 to 2 MDa) sharing several subunits but apparently differing in others include the TRAP/SMCC, NAT, DRIP, ARC, and human Mediator complexes. Smaller multiprotein complexes (approximately 500 to 700 kDa), including the murine Mediator, CRSP, and PC2, have also been described that contain subsets of subunits of the larger complexes. To evaluate whether these different multiprotein complexes exist in vivo in a single form or in multiple different forms, HeLa cell nuclear extract was directly resolved over a Superose 6 gel filtration column. Immunoblotting of column fractions using antisera specific for several Mediator subunits revealed one major size class of high-molecular-mass (approximately 2-MDa) complexes containing multiple mammalian Mediator subunits. No peak was apparent at approximately 500 to 700 kDa, indicating that either the smaller complexes reported are much less abundant than the higher-molecular-mass complexes or they are subcomplexes generated by dissociation of larger complexes during purification. Quantitative immunoblotting indicated that there are about 3 x 10(5) to 6 x 10(5) molecules of hSur2 Mediator subunit per HeLa cell, i.e., the same order of magnitude as RNA polymerase II and general transcription factors. Immunoprecipitation of the approximately 2-MDa fraction with anti-Cdk8 antibody indicated that at least two classes of Mediator complexes occur, one containing CDK8 and cyclin C and one lacking this CDK-cyclin pair. The approximately 2-MDa complexes stimulated activated transcription in vitro, whereas a 150-kDa fraction containing a subset of Mediator subunits inhibited activated transcription. PMID- 11416139 TI - Inhibition of androgen receptor-mediated transcription by amino-terminal enhancer of split. AB - A yeast two-hybrid assay has identified an androgen-dependent interaction of androgen receptor (AR) with amino-terminal enhancer of split (AES), a member of the highly conserved Groucho/TLE family of corepressors. Full-length AR, as well as the N-terminal fragment of AR, showed direct interactions with AES in in vitro protein-protein interaction assays. AES specifically inhibited AR-mediated transcription in a well-defined cell-free transcription system and interacted specifically with the basal transcription factor (TFIIE) in HeLa nuclear extract. These observations implicate AES as a selective repressor of ligand-dependent AR mediated transcription that acts by directly interacting with AR and by targeting the basal transcription machinery. PMID- 11416140 TI - Downregulation of CIITA function by protein kinase a (PKA)-mediated phosphorylation: mechanism of prostaglandin E, cyclic AMP, and PKA inhibition of class II major histocompatibility complex expression in monocytic lines. AB - Prostaglandins, pleiotropic immune modulators that induce protein kinase A (PKA), inhibit gamma interferon induction of class II major histocompatibility complex (MHC) genes. We show that phosphorylation of CIITA by PKA accounts for this inhibition. Treatment with prostaglandin E or 8-bromo-cyclic AMP or transfection with PKA inhibits the activity of CIITA in both mouse and human monocytic cell lines. This inhibition is independent of other transcription factors for the class II MHC promoter. These same treatments also greatly reduced the induction of class II MHC mRNA by CIITA. PKA phosphorylation sites were identified using site-directed mutagenesis and phosphoamino acid analysis. Phosphorylation at CIITA serines 834 and 1050 accounts for the inhibitory effects of PKA on CIITA driven class II MHC transcription. This is the first demonstration that the posttranslational modification of CIITA mediates inhibition of class II MHC transcription. PMID- 11416141 TI - CREB is one component of the binding complex of the Ces-2/E2A-HLF binding element and is an integral part of the interleukin-3 survival signal. AB - The Ces-2/E2A-HLF binding element (CBE) is recognized by Caenorhabditis elegans death specification gene product Ces-2 and human acute lymphocytic leukemia oncoprotein E2A-HLF. In an attempt to identify a cellular CBE-binding protein(s) that may be involved in apoptosis regulation in mammals, multiple nuclear binding complexes of CBE were identified in various mammalian cell lines and tissues by electrophoretic mobility shift assay. Cyclic AMP (cAMP)-responsive element (CRE) binding protein (CREB) was present in one major CBE complex of Ba/F3 and TF-1 cells, and both in vitro-translated and Escherichia coli-synthesized CREB bound to CBE. Activation of CREB by cAMP-elevating chemicals or the catalytic subunit of protein kinase A (PKAc) resulted in induction of the CBE-driven reporter gene. Stimulation of Ba/F3 cells with interleukin-3 (IL-3) promptly induced phosphorylation of CREB at serine(133) partially via a PKA-dependent pathway. Consistently, Ba/F3 cell survival in the absence of IL-3 was prolonged by activation of PKA. Conversely, treatment of cells with a PKA inhibitor or expression of the dominant negative forms of the regulatory subunit type I of PKA and CREB overrode the survival activity of IL-3. Last, the bcl-2 gene was demonstrated to be one candidate cellular target of the CREB-containing CBE complex, as mutations in the CRE and CBE sites significantly reduced the IL-3 inducibility of the bcl-2 promoter. Together, our results suggest that CREB is one cellular counterpart of Ces-2/E2A-HLF and is part of IL-3 dependent apoptosis regulation in hematopoietic cells. PMID- 11416142 TI - Bmx tyrosine kinase has a redundant function downstream of angiopoietin and vascular endothelial growth factor receptors in arterial endothelium. AB - The Bmx gene, a member of the Tec tyrosine kinase gene family, is known to be expressed in subsets of hematopoietic and endothelial cells. In this study, mice were generated in which the first coding exon of the Bmx gene was replaced with the lacZ reporter gene by a knock-in strategy. The homozygous mice lacking Bmx activity were fertile and had a normal life span without an obvious phenotype. Staining of their tissues using beta-galactosidase substrate to assess the sites of Bmx expression revealed strong signals in the endothelial cells of large arteries and in the endocardium starting between days 10.5 and 12.5 of embryogenesis and continuing in adult mice, while the venular endothelium showed a weak signal only in the superior and inferior venae cavae. Of the five known endothelial receptor tyrosine kinases tested, activated Tie-2 induced tyrosyl phosphorylation of the Bmx protein and both Tie-2 and vascular endothelial growth factor receptor 1 (VEGFR-1) stimulated Bmx tyrosine kinase activity. Thus, the Bmx tyrosine kinase has a redundant role in arterial endothelial signal transduction downstream of the Tie-2 and VEGFR-1 growth factor receptors. PMID- 11416143 TI - Mechanism of prion loss after Hsp104 inactivation in yeast. AB - In vivo propagation of [PSI(+)], an aggregation-prone prion isoform of the yeast release factor Sup35 (eRF3), has previously been shown to require intermediate levels of the chaperone protein Hsp104. Here we perform a detailed study on the mechanism of prion loss after Hsp104 inactivation. Complete or partial inactivation of Hsp104 was achieved by the following approaches: deleting the HSP104 gene; modifying the HSP104 promoter that results in low level of its expression; and overexpressing the dominant-negative ATPase-inactive mutant HSP104 allele. In contrast to guanidine-HCl, an agent blocking prion proliferation, Hsp104 inactivation induced relatively rapid loss of [PSI(+)] and another candidate yeast prion, [PIN(+)]. Thus, the previously hypothesized mechanism of prion dilution in cell divisions due to the blocking of prion proliferation is not sufficient to explain the effect of Hsp104 inactivation. The [PSI(+)] response to increased levels of another chaperone, Hsp70-Ssa, depends on whether the Hsp104 activity is increased or decreased. A decrease of Hsp104 levels or activity is accompanied by a decrease in the number of Sup35(PSI+) aggregates and an increase in their size. This eventually leads to accumulation of huge agglomerates, apparently possessing reduced prion forming capability and representing dead ends of the prion replication cycle. Thus, our data confirm that the primary function of Hsp104 in prion propagation is to disassemble prion aggregates and generate the small prion seeds that initiate new rounds of prion propagation (possibly assisted by Hsp70-Ssa). PMID- 11416144 TI - ZBP-89 promotes growth arrest through stabilization of p53. AB - Transcription factor p53 can induce growth arrest and/or apoptosis in cells through activation or repression of downstream target genes. Recently, we reported that ZBP-89 cooperates with histone acetyltransferase coactivator p300 in the regulation of p21(waf1), a cyclin-dependent kinase inhibitor whose associated gene is a target gene of p53. Therefore, we examined whether ZBP-89 might also inhibit cell growth by activating p53. In the present study, we demonstrate that elevated levels of ZBP-89 induce growth arrest and apoptosis in human gastrointestinal cell lines. The ZBP-89 protein accumulated within 4 h, and the p53 protein accumulated within 16 h, of serum starvation without changes in p14ARF levels, demonstrating a physiological increase in the cellular levels of these two proteins. Overexpression of ZBP-89 stabilized the p53 protein and enhanced its transcriptional activity through direct protein-protein interactions. The DNA binding and C-terminal domains of p53 and the zinc finger domain of ZBP-89 mediated the interaction. A point mutation in the p53 DNA binding domain, R273H, greatly reduced ZBP-89-mediated stabilization but not their physical interaction. Furthermore, ZBP-89 formed a complex with p53 and MDM2 and therefore did not prevent the MDM2-p53 interaction. However, heterokaryon assays demonstrated that ZBP-89 retained p53 in the nucleus. Collectively, these data indicate that ZBP-89 regulates cell proliferation in part through its ability to directly bind the p53 protein and retard its nuclear export. Our findings further our understanding of how ZBP-89 modulates cell proliferation and reveals a novel mechanism by which the p53 protein is stabilized. PMID- 11416145 TI - Role for E2F in control of both DNA replication and mitotic functions as revealed from DNA microarray analysis. AB - We have used high-density DNA microarrays to provide an analysis of gene regulation during the mammalian cell cycle and the role of E2F in this process. Cell cycle analysis was facilitated by a combined examination of gene control in serum-stimulated fibroblasts and cells synchronized at G(1)/S by hydroxyurea block that were then released to proceed through the cell cycle. The latter approach (G(1)/S synchronization) is critical for rigorously maintaining cell synchrony for unambiguous analysis of gene regulation in later stages of the cell cycle. Analysis of these samples identified seven distinct clusters of genes that exhibit unique patterns of expression. Genes tend to cluster within these groups based on common function and the time during the cell cycle that the activity is required. Placed in this context, the analysis of genes induced by E2F proteins identified genes or expressed sequence tags not previously described as regulated by E2F proteins; surprisingly, many of these encode proteins known to function during mitosis. A comparison of the E2F-induced genes with the patterns of cell growth-regulated gene expression revealed that virtually all of the E2F-induced genes are found in only two of the cell cycle clusters; one group was regulated at G(1)/S, and the second group, which included the mitotic activities, was regulated at G(2). The activation of the G(2) genes suggests a broader role for E2F in the control of both DNA replication and mitotic activities. PMID- 11416146 TI - DNA recognition by the herpes simplex virus transactivator VP16: a novel DNA binding structure. AB - Upon infection, the herpes simplex virus (HSV) transcriptional activator VP16 directs the formation of a multiprotein-DNA complex-the VP16-induced complex-with two cellular proteins, the host cell factor HCF-1 and the POU domain transcription factor Oct-1, on TAATGARAT-containing sequences found in the promoters of HSV immediate-early genes. HSV VP16 contains carboxy-terminal sequences important for transcriptional activation and a central conserved core that is important for VP16-induced complex assembly. On its own, VP16 displays little, if any, sequence-specific DNA-binding activity. We show here that, within the VP16-induced complex, however, the VP16 core has an important role in DNA binding. Mutation of basic residues on the surface of the VP16 core reveals a novel DNA-binding surface with essential residues which are conserved among VP16 orthologs. These results illuminate how, through association with DNA, VP16 is able to interpret cis-regulatory signals in the DNA to direct the assembly of a multiprotein-DNA transcriptional regulatory complex. PMID- 11416147 TI - The MLK family mediates c-Jun N-terminal kinase activation in neuronal apoptosis. AB - Neuronal apoptotic death induced by nerve growth factor (NGF) deprivation is reported to be in part mediated through a pathway that includes Rac1 and Cdc42, mitogen-activated protein kinase kinases 4 and 7 (MKK4 and -7), c-Jun N-terminal kinases (JNKs), and c-Jun. However, additional components of the pathway remain to be defined. We show here that members of the mixed-lineage kinase (MLK) family (including MLK1, MLK2, MLK3, and dual leucine zipper kinase [DLK]) are expressed in neuronal cells and are likely to act between Rac1/Cdc42 and MKK4 and -7 in death signaling. Overexpression of MLKs effectively induces apoptotic death of cultured neuronal PC12 cells and sympathetic neurons, while expression of dominant-negative forms of MLKs suppresses death evoked by NGF deprivation or expression of activated forms of Rac1 and Cdc42. CEP-1347 (KT7515), which blocks neuronal death caused by NGF deprivation and a variety of additional apoptotic stimuli and which selectively inhibits the activities of MLKs, effectively protects neuronal PC12 cells from death induced by overexpression of MLK family members. In addition, NGF deprivation or UV irradiation leads to an increase in both level and phosphorylation of endogenous DLK. These observations support a role for MLKs in the neuronal death mechanism. With respect to ordering the death pathway, dominant-negative forms of MKK4 and -7 and c-Jun are protective against death induced by MLK overexpression, placing MLKs upstream of these kinases. Additional findings place the MLKs upstream of mitochondrial cytochrome c release and caspase activation. PMID- 11416148 TI - Myc potentiates apoptosis by stimulating Bax activity at the mitochondria. AB - The ability of the c-Myc oncoprotein to potentiate apoptosis has been well documented; however, the mechanism of action remains ill defined. We have previously identified spatially distinct apoptotic pathways within the same cell that are differentially inhibited by Bcl-2 targeted to either the mitochondria (Bcl-acta) or the endoplasmic reticulum (Bcl-cb5). We show here that in Rat1 cells expressing an exogenous c-myc allele, distinct apoptotic pathways can be inhibited by Bcl-2 or Bcl-acta yet be distinguished by their sensitivity to Bcl cb5 as either susceptible (serum withdrawal, taxol, and ceramide) or refractory (etoposide and doxorubicin). Myc expression and apoptosis were universally associated with Bcl-acta and not Bcl-cb5, suggesting that Myc acts downstream at a point common to these distinct apoptotic signaling cascades. Analysis of Rat1 c myc null cells shows these same death stimuli induce apoptosis with characteristic features of nuclear condensation, membrane blebbing, poly (ADP ribose) polymerase cleavage, and DNA fragmentation; however, this Myc-independent apoptosis is not inhibited by Bcl-2. During apoptosis, Bax translocation to the mitochondria occurs in the presence or absence of Myc expression. Moreover, Bax mRNA and protein expression remain unchanged in the presence or absence of Myc. However, in the absence of Myc, Bax is not activated and cytochrome c is not released into the cytoplasm. Reintroduction of Myc into the c-myc null cells restores Bax activation, cytochrome c release, and inhibition of apoptosis by Bcl 2. These results demonstrate a role for Myc in the regulation of Bax activation during apoptosis. Moreover, apoptosis that can be triggered in the absence of Myc provides evidence that signaling pathways exist which circumvent Bax activation and cytochrome c release to trigger caspase activation. Thus, Myc increases the cellular competence to die by enhancing disparate apoptotic signals at a common mitochondrial amplification step involving Bax activation and cytochrome c release. PMID- 11416149 TI - Postrepression activation of NF-kappaB requires the amino-terminal nuclear export signal specific to IkappaBalpha. AB - One of the most prominent NF-kappaB target genes in mammalian cells is the gene encoding one of its inhibitor proteins, IkappaBalpha. The increased synthesis of IkappaBalpha leads to postinduction repression of nuclear NF-kappaB activity. However, it is unknown why IkappaBalpha, among multiple IkappaB family members, is involved in this process and what significance this feedback regulation has beyond terminating NF-kappaB activity. Herein, we report an important IkappaBalpha-specific function dictated by its amino-terminal nuclear export sequence (N-NES). The IkappaBalpha N-NES is necessary for the postinduction export of nuclear NF-kappaB, which is a critical event in reestablishing a permissive condition for NF-kappaB to be rapidly reactivated. We show that although IkappaBalpha and another IkappaB member, IkappaBbeta, can enter the nucleus and repress NF-kappaB DNA-binding activity during the postinduction phase, only IkappaBalpha allows the efficient export of nuclear NF-kappaB. Moreover, swapping the N-terminal region of IkappaBbeta for the corresponding IkappaBalpha sequence is sufficient for the IkappaB chimera protein to export NF kappaB similarly to IkappaBalpha during the postinduction state. Our findings provide a mechanistic explanation of why IkappaBalpha but not other IkappaB members is crucial for postrepression activation of NF-kappaB. We propose that this IkappaBalpha-specific function is important for certain physiological and pathological conditions where NF-kappaB needs to be rapidly reactivated. PMID- 11416150 TI - Genetic analysis reveals different functions for the products of the thyroid hormone receptor alpha locus. AB - Thyroid hormone receptors are encoded by the TRalpha (NR1A1) and TRbeta (NR1A2) loci. These genes are transcribed into multiple variants whose functions are unclear. Analysis by gene inactivation in mice has provided new insights into the functional complexity of these products. Different strategies designed to modify the TRalpha locus have led to strikingly different phenotypes. In order to analyze the molecular basis for these alterations, we generated mice devoid of all known isoforms produced from the TRalpha locus (TRalpha(0/0)). These mice are viable and exhibit reduced linear growth, bone maturation delay, moderate hypothermia, and reduced thickness of the intestinal mucosa. Compounding TRalpha(0) and TRbeta(-) mutations produces viable TRalpha(0/0)beta(-/-) mice, which display a more severe linear growth reduction and a more profound hypothermia as well as impaired hearing. A striking phenotypic difference is observed between TRalpha(0/0) and the previously described TRalpha(-/-) mice, which retain truncated TRDeltaalpha isoforms arising from a newly described promoter in intron 7. The lethality and severe impairment of the intestinal maturation in TRalpha(-/-) mice are rescued in TRalpha(0/0) animals. We demonstrate that the TRDeltaalpha protein isoforms, which are natural products of the TRalpha locus, are the key determinants of these phenotypical differences. These data reveal the functional importance of the non-T3-binding variants encoded by the TRalpha locus in vertebrate postnatal development and homeostasis. PMID- 11416151 TI - Functional interference between thyroid hormone receptor alpha (TRalpha) and natural truncated TRDeltaalpha isoforms in the control of intestine development. AB - Thyroid hormone is known to participate in the control of intestine maturation at weaning. Its action is mediated by the thyroid hormone nuclear receptors, encoded by the TRalpha and TRbeta genes. Since previous studies have shown that TRbeta plays a minor role in the gut, we focused here our analysis on the TRalpha gene. The TRalpha locus generates the TRalpha1 receptor together with the splicing variant TRalpha2 and the truncated products TRDeltaalpha1 and TRDeltaalpha2, which all lack an intact ligand binding domain. The TRDeltaalpha isoforms are transcribed from an internal promoter located in intron 7, and their distribution is restricted to a few tissues including those of the intestine. In order to define the functions of the different isoforms encoded by the TRalpha locus in the intestinal mucosa, we produced mice either lacking all known TRalpha products or harboring a mutation which inactivates the intronic promoter. We performed a detailed analysis of the intestinal phenotypes in these mice and compared it to that of the previously described TRalpha(-/-) mice, in which TRalpha isoforms are abolished but the TRDeltaalpha isoforms remain. This comparative analysis leads us to the following conclusions: (i) the TRalpha1 receptor mediates the T3 dependent functions in the intestine at weaning time and (ii) the TRDeltaalpha products negatively control the responsiveness of the epithelial cells to T3. Moreover, we show that TRDeltaalpha proteins can interfere with the transcription of the intestine-specific homeobox genes cdx1 and cdx2 and that their activity is regulated by TRalpha1. Altogether these data demonstrate that cooperation of TRalpha and TRDeltaalpha products is essential to ensure the normal postnatal development of the intestine and that mutations in the TRalpha locus can generate different phenotypes caused by the disruption of the equilibrium between these products. PMID- 11416152 TI - Differential regulation of retinoblastoma tumor suppressor protein by G(1) cyclin dependent kinase complexes in vivo. AB - The retinoblastoma tumor suppressor protein (pRB) negatively regulates early-G(1) cell cycle progression, in part, by sequestering E2F transcription factors and repressing E2F-responsive genes. Although pRB is phosphorylated on up to 16 cyclin-dependent kinase (Cdk) sites by multiple G(1) cyclin-Cdk complexes, the active form(s) of pRB in vivo remains unknown. pRB is present as an unphosphorylated protein in G(0) quiescent cells and becomes hypophosphorylated (approximately 2 mol of PO(4) to 1 mol of pRB) in early G(1) and hyperphosphorylated (approximately 10 mol of PO(4) to 1 mol of pRB) in late G(1) phase. Here, we report that hypophosphorylated pRB, present in early G(1), represents the biologically active form of pRB in vivo that is assembled with E2Fs and E1A but that both unphosphorylated pRB in G(0) and hyperphosphorylated pRB in late G(1) fail to become assembled with E2Fs and E1A. Furthermore, using transducible dominant-negative TAT fusion proteins that differentially target cyclin D-Cdk4 or cyclin D-Cdk6 (cyclin D-Cdk4/6) and cyclin E-Cdk2 complexes, namely, TAT-p16 and TAT-dominant-negative Cdk2, respectively, we found that, in vivo, cyclin D-Cdk4/6 complexes hypophosphorylate pRB in early G(1) and that cyclin E-Cdk2 complexes inactivate pRB by hyperphosphorylation in late G(1). Moreover, we found that cycling human tumor cells expressing deregulated cyclin D Cdk4/6 complexes, due to deletion of the p16(INK4a) gene, contained hypophosphorylated pRB that was bound to E2Fs in early G(1) and that E2F responsive genes, including those for dihydrofolate reductase and cyclin E, were transcriptionally repressed. Thus, we conclude that, physiologically, pRB is differentially regulated by G(1) cyclin-Cdk complexes. PMID- 11416153 TI - Insulin-responsive compartments containing GLUT4 in 3T3-L1 and CHO cells: regulation by amino acid concentrations. AB - In fat and muscle, insulin stimulates glucose uptake by rapidly mobilizing the GLUT4 glucose transporter from a specialized intracellular compartment to the plasma membrane. We describe a method to quantify the relative proportion of GLUT4 at the plasma membrane, using flow cytometry to measure a ratio of fluorescence intensities corresponding to the cell surface and total amounts of a tagged GLUT4 reporter in individual living cells. Using this assay, we demonstrate that both 3T3-L1 and CHO cells contain intracellular compartments from which GLUT4 is rapidly mobilized by insulin and that the initial magnitude and kinetics of redistribution to the plasma membrane are similar in these two cell types when they are cultured identically. Targeting of GLUT4 to a highly insulin-responsive compartment in CHO cells is modulated by culture conditions. In particular, we find that amino acids regulate distribution of GLUT4 to this kinetically defined compartment through a rapamycin-sensitive pathway. Amino acids also modulate the magnitude of insulin-stimulated translocation in 3T3-L1 adipocytes. Our results indicate a novel link between glucose and amino acid metabolism. PMID- 11416154 TI - Insulation of enhancer-promoter communication by a gypsy transposon insert in the Drosophila cut gene: cooperation between suppressor of hairy-wing and modifier of mdg4 proteins. AB - The Drosophila mod(mdg4) gene products counteract heterochromatin-mediated silencing of the white gene and help activate genes of the bithorax complex. They also regulate the insulator activity of the gypsy transposon when gypsy inserts between an enhancer and promoter. The Su(Hw) protein is required for gypsy mediated insulation, and the Mod(mdg4)-67.2 protein binds to Su(Hw). The aim of this study was to determine whether Mod(mdg4)-67.2 is a coinsulator that helps Su(Hw) block enhancers or a facilitator of activation that is inhibited by Su(Hw). Here we provide evidence that Mod(mdg4)-67.2 acts as a coinsulator by showing that some loss-of-function mod(mdg4) mutations decrease enhancer blocking by a gypsy insert in the cut gene. We find that the C terminus of Mod(mdg4)-67.2 binds in vitro to a region of Su(Hw) that is required for insulation, while the N terminus mediates self-association. The N terminus of Mod(mdg4)-67.2 also interacts with the Chip protein, which facilitates activation of cut. Mod(mdg4) 67.2 truncated in the C terminus interferes in a dominant-negative fashion with insulation in cut but does not significantly affect heterochromatin-mediated silencing of white. We infer that multiple contacts between Su(Hw) and a Mod(mdg4)-67.2 multimer are required for insulation. We theorize that Mod(mdg4) 67.2 usually aids gene activation but can also act as a coinsulator by helping Su(Hw) trap facilitators of activation, such as the Chip protein. PMID- 11416155 TI - The cell cycle-regulatory CDC25A phosphatase inhibits apoptosis signal-regulating kinase 1. AB - CDC25A phosphatase promotes cell cycle progression by activating G(1) cyclin dependent kinases and has been postulated to be an oncogene because of its ability to cooperate with RAS to transform rodent fibroblasts. In this study, we have identified apoptosis signal-regulating kinase 1 (ASK1) as a CDC25A interacting protein by yeast two-hybrid screening. ASK1 activates the p38 mitogen activated protein kinase (MAPK) and c-Jun NH(2)-terminal protein kinase-stress activated protein kinase (JNK/SAPK) pathways upon various cellular stresses. Coimmunoprecipitation studies demonstrated that CDC25A physically associates with ASK1 in mammalian cells, and immunocytochemistry with confocal laser-scanning microscopy showed that these two proteins colocalize in the cytoplasm. The carboxyl terminus of CDC25A binds to a domain of ASK1 adjacent to its kinase domain and inhibits the kinase activity of ASK1, independent of and without effect on the phosphatase activity of CDC25A. This inhibitory action of CDC25A on ASK1 activity involves diminished homo-oligomerization of ASK1. Increased cellular expression of wild-type or phosphatase-inactive CDC25A from inducible transgenes suppresses oxidant-dependent activation of ASK1, p38, and JNK1 and reduces specific sensitivity to cell death triggered by oxidative stress, but not other apoptotic stimuli. Thus, increased expression of CDC25A, frequently observed in human cancers, could contribute to reduced cellular responsiveness to oxidative stress under mitogenic or oncogenic conditions, while it promotes cell cycle progression. These observations propose a mechanism of oncogenic transformation by the dual function of CDC25A on cell cycle progression and stress responses. PMID- 11416156 TI - Proteinuria and perinatal lethality in mice lacking NEPH1, a novel protein with homology to NEPHRIN. AB - A high-throughput, retrovirus-mediated mutagenesis method based on gene trapping in embryonic stem cells was used to identify a novel mouse gene. The human ortholog encodes a transmembrane protein containing five extracellular immunoglobulin-like domains that is structurally related to human NEPHRIN, a protein associated with congenital nephrotic syndrome. Northern analysis revealed wide expression in humans and mice, with highest expression in kidney. Based on similarity to NEPHRIN and abundant expression in kidney, this protein was designated NEPH1 and embryonic stem cells containing the retroviral insertion in the Neph1 locus were used to generate mutant mice. Analysis of kidney RNA from Neph1(-/-) mice showed that the retroviral insertion disrupted expression of Neph1 transcripts. Neph1(-/-) pups were represented at the expected normal Mendelian ratios at 1 to 3 days of age but at only 10% of the expected frequency at 10 to 12 days after birth, suggesting an early postnatal lethality. The Neph1( /-) animals that survived beyond the first week of life were sickly and small but without edema, and all died between 3 and 8 weeks of age. Proteinuria ranging from 300 to 2,000 mg/dl was present in all Neph1(-/-) mice. Electron microscopy demonstrated NEPH1 expression in glomerular podocytes and revealed effacement of podocyte foot processes in Neph1(-/-) mice. These findings suggest that NEPH1, like NEPHRIN, may play an important role in maintaining the structure of the filtration barrier that prevents proteins from freely entering the glomerular urinary space. PMID- 11416157 TI - Cell-specific association and shuttling of IkappaBalpha provides a mechanism for nuclear NF-kappaB in B lymphocytes. AB - Mature B lymphocytes are unique in containing nuclear Rel proteins prior to cell stimulation. This activity consists largely of p50-c-Rel heterodimers, and its importance for B-cell function is exemplified by reduced B-cell viability in several genetically altered mouse strains. Here we suggest a mechanism for the cell specificity and the subunit composition of constitutive B-cell NF-kappaB based on the observed properties of Rel homo- and heterodimers and IkappaBalpha. We show that c-Rel lacks a nuclear export sequence, making the removal of c-Rel containing complexes from the nucleus less efficient than removal of p65 containing complexes. Second, the nuclear import potential of p65 and c-Rel homodimers but not p50-associated heterodimers was attenuated when they were complexed to IkappaBalpha, leading to a greater propensity of heterodimers to be nuclear. We propose that subunit composition of B-cell NF-kappaB reflects the inefficient retrieval of p50-c-Rel heterodimers from the nucleus. Cell specificity may be a consequence of c-Rel-IkappaBalpha complexes being present only in mature B cells, which leads to nuclear c-Rel due to IkappaBalpha turnover and shuttling of the complex. PMID- 11416158 TI - Hd6, a rice quantitative trait locus involved in photoperiod sensitivity, encodes the alpha subunit of protein kinase CK2. AB - Hd6 is a quantitative trait locus involved in rice photoperiod sensitivity. It was detected in backcross progeny derived from a cross between the japonica variety Nipponbare and the indica variety Kasalath. To isolate a gene at Hd6, we used a large segregating population for the high-resolution and fine-scale mapping of Hd6 and constructed genomic clone contigs around the Hd6 region. Linkage analysis with P1-derived artificial chromosome clone-derived DNA markers delimited Hd6 to a 26.4-kb genomic region. We identified a gene encoding the alpha subunit of protein kinase CK2 (CK2 alpha) in this region. The Nipponbare allele of CK2 alpha contains a premature stop codon, and the resulting truncated product is undoubtedly nonfunctional. Genetic complementation analysis revealed that the Kasalath allele of CK2 alpha increases days-to-heading. Map-based cloning with advanced backcross progeny enabled us to identify a gene underlying a quantitative trait locus even though it exhibited a relatively small effect on the phenotype. PMID- 11416159 TI - Short tandem repeat profiling provides an international reference standard for human cell lines. AB - Cross-contamination between cell lines is a longstanding and frequent cause of scientific misrepresentation. Estimates from national testing services indicate that up to 36% of cell lines are of a different origin or species to that claimed. To test a standard method of cell line authentication, 253 human cell lines from banks and research institutes worldwide were analyzed by short tandem repeat profiling. The short tandem repeat profile is a simple numerical code that is reproducible between laboratories, is inexpensive, and can provide an international reference standard for every cell line. If DNA profiling of cell lines is accepted and demanded internationally, scientific misrepresentation because of cross-contamination can be largely eliminated. PMID- 11416160 TI - Expression and functional significance of VE-cadherin in aggressive human melanoma cells: role in vasculogenic mimicry. AB - We recently have introduced the term vasculogenic mimicry to describe the unique ability of aggressive melanoma tumor cells to form tubular structures and patterned networks in three-dimensional culture, which "mimics" embryonic vasculogenic networks formed by differentiating endothelial cells. In the current study, we address the biological significance of several endothelial-associated molecules (revealed by microarray analysis) with respect to expression and function in highly aggressive and poorly aggressive human cutaneous melanoma cell lines (established from the same patient). In a comparative analysis, CD31 was not expressed by any of the melanoma cell lines, whereas TIE-1 (tyrosine kinase with Ig and epidermal growth factor homology domains-1) was strongly expressed in the highly aggressive tumor cells with a low level of expression in one of the poorly aggressive cell lines. Vascular endothelial (VE)-cadherin was exclusively expressed by highly aggressive melanoma cells and was undetectable in the poorly aggressive tumor cells, suggesting the possibility of a vasculogenic switch. Down regulation of VE-cadherin expression in the aggressive melanoma cells abrogated their ability to form vasculogenic networks and directly tested the hypothesis that VE-cadherin is critical in melanoma vasculogenic mimicry. These results highlight the plasticity of aggressive melanoma cells and call into question their possible genetic reversion to an embryonic phenotype. This finding could pose a significant clinical challenge in targeting tumor cells that may masquerade as circulating endothelial cells or other embryonic-like stem cells. PMID- 11416161 TI - Adenosine diphosphate sugar pyrophosphatase prevents glycogen biosynthesis in Escherichia coli. AB - An adenosine diphosphate sugar pyrophosphatase (ASPPase, EC ) has been characterized by using Escherichia coli. This enzyme, whose activities in the cell are inversely correlated with the intracellular glycogen content and the glucose concentration in the culture medium, hydrolyzes ADP-glucose, the precursor molecule of glycogen biosynthesis. ASPPase was purified to apparent homogeneity (over 3,000-fold), and sequence analyses revealed that it is a member of the ubiquitously distributed group of nucleotide pyrophosphatases designated as "nudix" hydrolases. Insertional mutagenesis experiments leading to the inactivation of the ASPPase encoding gene, aspP, produced cells with marginally low enzymatic activities and higher glycogen content than wild-type bacteria. aspP was cloned into an expression vector and introduced into E. coli. Transformed cells were shown to contain a dramatically reduced amount of glycogen, as compared with the untransformed bacteria. No pleiotropic changes in the bacterial growth occurred in both the aspP-overexpressing and aspP-deficient strains. The overall results pinpoint the reaction catalyzed by ASPPase as a potential step of regulating glycogen biosynthesis in E. coli. PMID- 11416162 TI - Increased sexual activity reduces male immune function in Drosophila melanogaster. AB - Despite the benefits of resistance, susceptibility to infectious disease is commonplace. Although specific susceptibility may be considered an inevitable consequence of the co-evolutionary arms race between parasite and host, a more general constraint may arise from the cost of an immune response. This "cost" hypothesis predicts a tradeoff between immune defense and other components of fitness. In particular, a tradeoff between immunity and sexually selected male behavior has been proposed. Here we provide experimental support for the direct phenotypic tradeoff between sexual activity and immunity by studying the antibacterial immune response in Drosophila melanogaster. Males exposed to more females showed a reduced ability to clear a bacterial infection, an effect that we experimentally link to changes in sexual activity. Our results suggest immunosuppression is an important cost of reproduction and that immune function and levels of disease susceptibility will be influenced by sexual selection. PMID- 11416163 TI - The type I BMP receptor BmprIB is essential for female reproductive function. AB - Maintenance of female reproductive competence depends on the actions of several hormones and signaling factors. Recent reports suggest roles for bone morphogenetic proteins (BMPs) in early stages of folliculogenesis. A role for the type I BMP receptor BmprIB as a regulator of ovulation rates in sheep has been described recently, but little is known about the roles of BMP signaling pathways in other aspects of reproductive function. We report here that BMPRIB is essential for multiple aspects of female fertility. Mice deficient in BmprIB exhibit irregular estrous cycles and an impaired pseudopregnancy response. BmprIB mutants produce oocytes that can be fertilized in vitro, but defects in cumulus expansion prevent fertilization in vivo. This defect is associated with decreased levels of aromatase production in granulosa cells. Unexpectedly, levels of mRNA for cyclooxygenase 2, an enzyme required for cumulus expansion, are increased. BmprIB mutants also exhibit a failure in endometrial gland formation. The expression of BmprIB in uterine linings suggests that these defects are a direct consequence of loss of BMP signaling in this tissue. In summary, these studies demonstrate the importance of BMP signaling pathways for estrus cyclicity, estradiol biosynthesis, and cumulus cell expansion in vivo and reveal sites of action for BMP signaling pathways in reproductive tissues. PMID- 11416164 TI - GTP-binding protein beta gamma subunits mediate presynaptic calcium current inhibition by GABA(B) receptor. AB - A variety of GTP-binding protein (G protein)-coupled receptors are expressed at the nerve terminals of central synapses and play modulatory roles in transmitter release. At the calyx of Held, a rat auditory brainstem synapse, activation of presynaptic gamma-aminobutyric acid type B receptors (GABA(B) receptors) or metabotropic glutamate receptors inhibits presynaptic P/Q-type Ca(2+) channel currents via activation of G proteins, thereby attenuating transmitter release. To identify the heterotrimeric G protein subunits involved in this presynaptic inhibition, we loaded G protein beta gamma subunits (G beta gamma) directly into the calyceal nerve terminal through whole-cell patch pipettes. G beta gamma slowed the activation of presynaptic Ca(2+) currents (I(pCa)) and attenuated its amplitude in a manner similar to the externally applied baclofen, a GABA(B) receptor agonist. The effects of both G beta gamma and baclofen were relieved after strong depolarization of the nerve terminal. In addition, G beta gamma partially occluded the inhibitory effect of baclofen on I(pCa). In contrast, guanosine 5'-O-(3-thiotriphosphate)-bound G(o)alpha loaded into the calyx had no effect. Immunocytochemical examination revealed that the subtype of G proteins G(o), but not the G(i), subtype, is expressed in the calyceal nerve terminal. These results suggest that presynaptic inhibition mediated by G protein-coupled receptors occurs primarily by means of the direct interaction of G(o) beta gamma subunits with presynaptic Ca(2+) channels. PMID- 11416165 TI - Maternal effects and the evolution of aposematic signals. AB - Aposematic signals that warn predators of the noxious qualities of prey gain their greatest selective advantage when predators have already experienced similar signals. Existing theory explains how such signals can spread through selective advantage after they are present at some critical frequency, but is unclear about how warning signals can be selectively advantageous when the trait is initially rare (i.e., when it first arises through mutation) and predators are naive. When aposematism is controlled by a maternal effect gene, the difficulty of initial rarity may be overcome. Unlike a zygotically expressed gene, a maternally expressed aposematism gene will be hidden from selection because it is not phenotypically expressed in the first individual with the mutation. Furthermore, the first individual carrying the new mutation will produce an entire family of aposematic offspring, thereby providing an immediate fitness advantage to this gene. PMID- 11416166 TI - Hydroperoxide lyase depletion in transgenic potato plants leads to an increase in aphid performance. AB - Hydroperoxide lyases (HPLs) catalyze the cleavage of fatty acid hydroperoxides to aldehydes and oxoacids. These volatile aldehydes play a major role in forming the aroma of many plant fruits and flowers. In addition, they have antimicrobial activity in vitro and thus are thought to be involved in the plant defense response against pest and pathogen attack. An HPL activity present in potato leaves has been characterized and shown to cleave specifically 13-hydroperoxides of both linoleic and linolenic acids to yield hexanal and 3-hexenal, respectively, and 12-oxo-dodecenoic acid. A cDNA encoding this HPL has been isolated and used to monitor gene expression in healthy and mechanically damaged potato plants. HPL gene expression is subject to developmental control, being high in young leaves and attenuated in older ones, and it is induced weakly by wounding. HPL enzymatic activity, nevertheless, remains constant in leaves of different ages and also after wounding, suggesting that posttranscriptional mechanisms may regulate its activity levels. Antisense-mediated HPL depletion in transgenic potato plants has identified this enzyme as a major route of 13-fatty acid hydroperoxide degradation in the leaves. Although these transgenic plants have highly reduced levels of both hexanal and 3-hexenal, they show no phenotypic differences compared with wild-type ones, particularly in regard to the expression of wound-induced genes. However, aphids feeding on the HPL-depleted plants display approximately a two-fold increase in fecundity above those feeding on nontransformed plants, consistent with the hypothesis that HPL-derived products have a negative impact on aphid performance. Thus, HPL-catalyzed production of C6 aldehydes may be a key step of a built-in resistance mechanism of plants against some sucking insect pests. PMID- 11416167 TI - Substrate-specific regulation of the ribosome- translocon junction by N-terminal signal sequences. AB - Amino-terminal signal sequences target nascent secretory and membrane proteins to the endoplasmic reticulum for translocation. Subsequent interactions between the signal sequence and components of the translocation machinery at the endoplasmic reticulum are thought to be important for the productive engagement of the translocon by the ribosome-nascent chain complex. However, it is not clear whether all signal sequences carry out these posttargeting steps identically, or if there are differences in the interactions directed by one signal sequence versus another. In this study, we find substantial differences in the ability of signal sequences from different substrates to mediate closure of the ribosome- translocon junction early in translocation. We also show that these differences in some cases necessitate functional coordination between the signal sequence and mature domain for faithful translocation. Accordingly, the translocation of some proteins is sensitive to replacement of their signal sequences. In a particularly dramatic example, the topology of the prion protein was found to depend highly on the choice of signal sequence used to direct its translocation. Taken together, our results reveal an unanticipated degree of substrate-specific functionality encoded in N-terminal signal sequences. PMID- 11416168 TI - The affective component of pain in rodents: direct evidence for a contribution of the anterior cingulate cortex. AB - Numerous human and animal studies indirectly implicate neurons in the anterior cingulate cortex (ACC) in the encoding of the affective consequences of nociceptor stimulation. No causal evidence, however, has been put forth linking the ACC specifically to this function. Using a rodent pain assay that combines the hind-paw formalin model with the place-conditioning paradigm, we measured a learned behavior that directly reflects the affective component of pain in the rat (formalin-induced conditioned place avoidance) concomitantly with "acute" formalin-induced nociceptive behaviors (paw lifting, licking, and flinching) that reflect the intensity and localization of the nociceptive stimulus. Destruction of neurons originating from the rostral, but not caudal, ACC reduced formalin induced conditioned place avoidance without reducing acute pain-related behaviors. These results provide evidence indicating that neurons in the ACC are necessary for the "aversiveness" of nociceptor stimulation. PMID- 11416169 TI - Nuclear particles containing RNA polymerase III complexes associated with the junctional plaque protein plakophilin 2. AB - Plakophilin 2, a member of the arm-repeat protein family, is a dual location protein that occurs both in the cytoplasmic plaques of desmosomes as an architectural component and in an extractable form in the nucleoplasm. Here we report the existence of two nuclear particles containing plakophilin 2 and the largest subunit of RNA polymerase (pol) III (RPC155), both of which colocalize and are coimmunoselected with other pol III subunits and with the transcription factor TFIIIB. We also show that plakophilin 2 is present in the pol III holoenzyme, but not the core complex, and that it binds specifically to RPC155 in vitro. We propose the existence of diverse nuclear particles in which proteins known as plaque proteins of intercellular junctions are complexed with specific nuclear proteins. PMID- 11416170 TI - Related homing endonucleases I-BmoI and I-TevI use different strategies to cleave homologous recognition sites. AB - A typical homing endonuclease initiates mobility of its group I intron by recognizing DNA both upstream and downstream of the intron insertion site of intronless alleles, preventing the endonuclease from binding and cleaving its own intron-containing allele. Here, we describe a GIY-YIG family homing endonuclease, I-BmoI, that possesses an unusual recognition sequence, encompassing 1 base pair upstream but 38 base pairs downstream of the intron insertion site. I-BmoI binds intron-containing and intronless substrates with equal affinity but can nevertheless discriminate between the two for cleavage. I-BmoI is encoded by a group I intron that interrupts the thymidylate synthase (TS) gene (thyA) of Bacillus mojavensis s87-18. This intron resembles one inserted 21 nucleotides further downstream in a homologous TS gene (td) of Escherichia coli phage T4. I TevI, the T4 td intron-encoded GIY-YIG endonuclease, is very similar to I-BmoI, but each endonuclease gene is inserted within a different position of its respective intron. Remarkably, I-TevI and I-BmoI bind a homologous stretch of TS encoding DNA and cleave their intronless substrates in very similar positions. Our results suggest that each endonuclease has independently evolved the ability to distinguish intron-containing from intronless alleles while maintaining the same conserved recognition sequence centered on DNA-encoding active site residues of TS. PMID- 11416171 TI - Redox-dependent activation of CO dehydrogenase from Rhodospirillum rubrum. AB - Studies of initial activities of carbon monoxide dehydrogenase (CODH) from Rhodospirillum rubrum show that CODH is mostly inactive at redox potentials higher than -300 mV. Initial activities measured at a wide range of redox potentials (0--500 mV) fit a function corresponding to the Nernst equation with a midpoint potential of -316 mV. Previously, extensive EPR studies of CODH have suggested that CODH has three distinct redox states: (i) a spin-coupled state at 60 to -300 mV that gives rise to an EPR signal termed C(red1); (ii) uncoupled states at <-320 mV in the absence of CO(2) referred to as C(unc); and (iii) another spin-coupled state at <-320 mV in the presence of CO(2) that gives rise to an EPR signal termed C(red2B). Because there is no initial CODH activity at potentials that give rise to C(red1), the state (C(red1)) is not involved in the catalytic mechanism of this enzyme. At potentials more positive than -380 mV, CODH recovers its full activity over time when incubated with CO. This reductant dependent conversion of CODH from an inactive to an active form is referred to hereafter as "autocatalysis." Analyses of the autocatalytic activation process of CODH suggest that the autocatalysis is initiated by a small fraction of activated CODH; the small fraction of active CODH catalyzes CO oxidation and consequently lowers the redox potential of the assay system. This process is accelerated with time because of accumulation of the active enzyme. PMID- 11416172 TI - Protection from superoxide damage associated with an increased level of the YggX protein in Salmonella enterica. AB - The deleterious effect of superoxide radicals on cell growth and survival is predominantly caused by rapid oxidation of labile [Fe-S] clusters in proteins. Oxidation of these clusters releases Fe(II) ions, which participate in Fenton chemistry that damages DNA. Here it is shown that elevated levels of the YggX protein increase the resistance of Salmonella enterica to superoxide stress, reverse enzymatic defects attributed to oxidized [Fe-S] clusters, and decrease the spontaneous mutation frequency. The data are consistent with a model in which YggX protects protein [Fe-S] clusters from oxidation. PMID- 11416173 TI - Rolling back to BOULE. PMID- 11416174 TI - DNA structure from A to B. PMID- 11416175 TI - Signaling cascades: escape from kinetic proofreading. PMID- 11416176 TI - Molecular matchmaking: NNRTIs can enhance the dimerization of HIV type 1 reverse transcriptase. PMID- 11416177 TI - Molecular kinesis in cellular function and plasticity. AB - Intracellular transport and localization of cellular components are essential for the functional organization and plasticity of eukaryotic cells. Although the elucidation of protein transport mechanisms has made impressive progress in recent years, intracellular transport of RNA remains less well understood. The National Academy of Sciences Colloquium on Molecular Kinesis in Cellular Function and Plasticity therefore was devised as an interdisciplinary platform for participants to discuss intracellular molecular transport from a variety of different perspectives. Topics covered at the meeting included RNA metabolism and transport, mechanisms of protein synthesis and localization, the formation of complex interactive protein ensembles, and the relevance of such mechanisms for activity-dependent regulation and synaptic plasticity in neurons. It was the overall objective of the colloquium to generate momentum and cohesion for the emerging research field of molecular kinesis. PMID- 11416178 TI - Kinesin molecular motors: transport pathways, receptors, and human disease. AB - Kinesin molecular motor proteins are responsible for many of the major microtubule-dependent transport pathways in neuronal and non-neuronal cells. Elucidating the transport pathways mediated by kinesins, the identity of the cargoes moved, and the nature of the proteins that link kinesin motors to cargoes are areas of intense investigation. Kinesin-II recently was found to be required for transport in motile and nonmotile cilia and flagella where it is essential for proper left-right determination in mammalian development, sensory function in ciliated neurons, and opsin transport and viability in photoreceptors. Thus, these pathways and proteins may be prominent contributors to several human diseases including ciliary dyskinesias, situs inversus, and retinitis pigmentosa. Kinesin-I is needed to move many different types of cargoes in neuronal axons. Two candidates for receptor proteins that attach kinesin-I to vesicular cargoes were recently found. One candidate, sunday driver, is proposed to both link kinesin-I to an unknown vesicular cargo and to bind and organize the mitogen activated protein kinase components of a c-Jun N-terminal kinase signaling module. A second candidate, amyloid precursor protein, is proposed to link kinesin-I to a different, also unknown, class of axonal vesicles. The finding of a possible functional interaction between kinesin-I and amyloid precursor protein may implicate kinesin-I based transport in the development of Alzheimer's disease. PMID- 11416179 TI - All kinesin superfamily protein, KIF, genes in mouse and human. AB - Intracellular transport is essential for morphogenesis and functioning of the cell. The kinesin superfamily proteins (KIFs) have been shown to transport membranous organelles and protein complexes in a microtubule- and ATP-dependent manner. More than 30 KIFs have been reported in mice. However, the nomenclature of KIFs has not been clearly established, resulting in various designations and redundant names for a single KIF. Here, we report the identification and classification of all KIFs in mouse and human genome transcripts. Previously unidentified murine KIFs were found by a PCR-based search. The identification of all KIFs was confirmed by a database search of the total human genome. As a result, there are a total of 45 KIFs. The nomenclature of all KIFs is presented. To understand the function of KIFs in intracellular transport in a single tissue, we focused on the brain. The expression of 38 KIFs was detected in brain tissue by Northern blotting or PCR using cDNA. The brain, mainly composed of highly differentiated and polarized cells such as neurons and glia, requires a highly complex intracellular transport system as indicated by the increased number of KIFs for their sophisticated functions. It is becoming increasingly clear that the cell uses a number of KIFs and tightly controls the direction, destination, and velocity of transportation of various important functional molecules, including mRNA. This report will set the foundation of KIF and intracellular transport research. PMID- 11416180 TI - Assembly and transport of a premessenger RNP particle. AB - Salivary gland cells in the larvae of the dipteran Chironomus tentans offer unique possibilities to visualize the assembly and nucleocytoplasmic transport of a specific transcription product. Each nucleus harbors four giant polytene chromosomes, whose transcription sites are expanded, or puffed. On chromosome IV, there are two puffs of exceptional size, Balbiani ring (BR) 1 and BR 2. A BR gene is 35-40 kb, contains four short introns, and encodes a 1-MDa salivary polypeptide. The BR transcript is packed with proteins into a ribonucleoprotein (RNP) fibril that is folded into a compact ring-like structure. The completed RNP particle is released into the nucleoplasm and transported to the nuclear pore, where the RNP fibril is gradually unfolded and passes through the pore. On the cytoplasmic side, the exiting extended RNP fibril becomes engaged in protein synthesis and the ensuing polysome is anchored to the endoplasmic reticulum. Several of the BR particle proteins have been characterized, and their fate during the assembly and transport of the BR particle has been elucidated. The proteins studied are all added cotranscriptionally to the pre-mRNA molecule. The various proteins behave differently during RNA transport, and the flow pattern of each protein is related to the particular function of the protein. Because the cotranscriptional assembly of the pre-mRNP particle involves proteins functioning in the nucleus as well as proteins functioning in the cytoplasm, it is concluded that the fate of the mRNA molecule is determined to a considerable extent already at the gene level. PMID- 11416181 TI - Ribonucleoprotein infrastructure regulating the flow of genetic information between the genome and the proteome. AB - Following transcription and splicing, each mRNA of a mammalian cell passes into the cytoplasm where its fate is in the hands of a complex network of ribonucleoproteins (mRNPs). The success or failure of a gene to be expressed depends on the performance of this mRNP infrastructure. The entry, gating, processing, and transit of each mRNA through an mRNP network helps determine the composition of a cell's proteome. The machinery that regulates storage, turnover, and translational activation of mRNAs is not well understood, in part, because of the heterogeneous nature of mRNPs. Recently, subsets of cellular mRNAs clustered as members of mRNP complexes have been identified by using antibodies reactive with RNA-binding proteins, including ELAV/Hu, eIF-4E, and poly(A)-binding proteins. Cytoplasmic ELAV/Hu proteins are involved in the stability and translation of early response gene (ERG) transcripts and are expressed predominately in neurons. mRNAs recovered from ELAV/Hu mRNP complexes were found to have similar sequence elements, suggesting a common structural linkage among them. This approach opens the possibility of identifying transcripts physically clustered in vivo that may have similar fates or functions. Moreover, the proteins encoded by physically organized mRNAs may participate in the same biological process or structural outcome, not unlike operons and their polycistronic mRNAs do in prokaryotic organisms. Our goal is to understand the organization and flow of genetic information on an integrative systems level by analyzing the collective properties of proteins and mRNAs associated with mRNPs in vivo. PMID- 11416182 TI - Spatial and temporal control of RNA stability. AB - Maternally encoded RNAs and proteins program the early development of all animals. A subset of the maternal transcripts is eliminated from the embryo before the midblastula transition. In certain cases, transcripts are protected from degradation in a subregion of the embryonic cytoplasm, thus resulting in transcript localization. Maternal factors are sufficient for both the degradation and protection components of transcript localization. Cis-acting elements in the RNAs convert transcripts progressively (i) from inherently stable to unstable and (ii) from uniformly degraded to locally protected. Similar mechanisms are likely to act later in development to restrict certain classes of transcripts to particular cell types within somatic cell lineages. Functions of transcript degradation and protection are discussed. PMID- 11416183 TI - Molecular mechanisms of translation initiation in eukaryotes. AB - Translation initiation is a complex process in which initiator tRNA, 40S, and 60S ribosomal subunits are assembled by eukaryotic initiation factors (eIFs) into an 80S ribosome at the initiation codon of mRNA. The cap-binding complex eIF4F and the factors eIF4A and eIF4B are required for binding of 43S complexes (comprising a 40S subunit, eIF2/GTP/Met-tRNAi and eIF3) to the 5' end of capped mRNA but are not sufficient to promote ribosomal scanning to the initiation codon. eIF1A enhances the ability of eIF1 to dissociate aberrantly assembled complexes from mRNA, and these factors synergistically mediate 48S complex assembly at the initiation codon. Joining of 48S complexes to 60S subunits to form 80S ribosomes requires eIF5B, which has an essential ribosome-dependent GTPase activity and hydrolysis of eIF2-bound GTP induced by eIF5. Initiation on a few mRNAs is cap independent and occurs instead by internal ribosomal entry. Encephalomyocarditis virus (EMCV) and hepatitis C virus epitomize distinct mechanisms of internal ribosomal entry site (IRES)-mediated initiation. The eIF4A and eIF4G subunits of eIF4F bind immediately upstream of the EMCV initiation codon and promote binding of 43S complexes. EMCV initiation does not involve scanning and does not require eIF1, eIF1A, and the eIF4E subunit of eIF4F. Initiation on some EMCV-like IRESs requires additional noncanonical initiation factors, which alter IRES conformation and promote binding of eIF4A/4G. Initiation on the hepatitis C virus IRES is even simpler: 43S complexes containing only eIF2 and eIF3 bind directly to the initiation codon as a result of specific interaction of the IRES and the 40S subunit. PMID- 11416184 TI - The target of rapamycin (TOR) proteins. AB - Rapamycin potently inhibits downstream signaling from the target of rapamycin (TOR) proteins. These evolutionarily conserved protein kinases coordinate the balance between protein synthesis and protein degradation in response to nutrient quality and quantity. The TOR proteins regulate (i) the initiation and elongation phases of translation, (ii) ribosome biosynthesis, (iii) amino acid import, (iv) the transcription of numerous enzymes involved in multiple metabolic pathways, and (v) autophagy. Intriguingly, recent studies have also suggested that TOR signaling plays a critical role in brain development, learning, and memory formation. PMID- 11416185 TI - The physiological significance of beta -actin mRNA localization in determining cell polarity and directional motility. AB - beta-actin mRNA is localized near the leading edge in several cell types, where actin polymerization is actively promoting forward protrusion. The localization of the beta-actin mRNA near the leading edge is facilitated by a short sequence in the 3' untranslated region, the "zip code." Localization of the mRNA at this region is important physiologically. Treatment of chicken embryo fibroblasts with antisense oligonucleotides complementary to the localization sequence (zip code) in the 3' untranslated region leads to delocalization of beta-actin mRNA, alteration of cell phenotype, and a decrease in cell motility. To determine the components of this process responsible for the change in cell behavior after beta actin mRNA delocalization, the Dynamic Image Analysis System was used to quantify movement of cells in the presence of sense and antisense oligonucleotides to the zip code. It was found that net path length and average speed of antisense treated cells were significantly lower than in sense-treated cells. Total path length and the velocity of protrusion of antisense-treated cells were not affected compared with those of control cells. These results suggest that a decrease in persistence of direction of movement and not in velocity results from treatment of cells with zip code-directed antisense oligonucleotides. To test this, direct analysis of directionality was performed on antisense-treated cells and showed a decrease in directionality (net path/total path) and persistence of movement. Less directional movement of antisense-treated cells correlated with a unpolarized and discontinuous distribution of free barbed ends of actin filaments and of beta-actin protein. These results indicate that delocalization of beta actin mRNA results in delocalization of nucleation sites and beta-actin protein from the leading edge followed by loss of cell polarity and directional movement. PMID- 11416186 TI - Sorting and directed transport of membrane proteins during development of hippocampal neurons in culture. AB - Hippocampal neurons in culture develop morphological polarity in a sequential pattern; axons form before dendrites. Molecular differences, particularly those of membrane proteins, underlie the functional polarity of these domains, yet little is known about the temporal relationship between membrane protein polarization and morphological polarization. We took advantage of viral expression systems to determine when during development the polarization of membrane proteins arises. All markers were unpolarized in neurons before axonogenesis. In neurons with a morphologically distinguishable axon, even on the first day in culture, both axonal and dendritic proteins were polarized. The degree of polarization at these early stages was somewhat less than in mature cells and varied from cell to cell. The cellular mechanism responsible for the polarization of the dendritic marker protein transferrin receptor (TfR) in mature cells centers on directed transport to the dendritic domain. To examine the relationship between cell surface polarization and transport, we assessed the selectivity of transport by live cell imaging. TfR-green fluorescent protein containing vesicles were already preferentially transported into dendrites at 2 days, the earliest time point we could measure. The selectivity of transport also varied somewhat among cells, and the amount of TfR-green fluorescent protein fluorescence on intracellular structures within the axon correlated with the amount of cell surface expression. This observation implies that selective microtubule-based transport is the primary mechanism that underlies the polarization of TfR on the cell surface. By 5 days in culture, the extent of polarization on the cell surface and the selectivity of transport reached mature levels. PMID- 11416187 TI - Molecular organization of the postsynaptic specialization. AB - A specific set of molecules including glutamate receptors is targeted to the postsynaptic specialization of excitatory synapses in the brain, gathering in a structure known as the postsynaptic density (PSD). Synaptic targeting of glutamate receptors depends on interactions between the C-terminal tails of receptor subunits and specific PDZ domain-containing scaffold proteins in the PSD. These scaffold proteins assemble a specialized protein complex around each class of glutamate receptor that functions in signal transduction, cytoskeletal anchoring, and trafficking of the receptors. Among the glutamate receptor subtypes, the N-methyl-d-aspartate receptor is relatively stably integrated in the PSD, whereas the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor moves in and out of the postsynaptic membrane in highly dynamic fashion. The distinctive cell biological behaviors of N-methyl-d-aspartate and alpha-amino 3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors can be explained by their differential interactions with cytoplasmic proteins. PMID- 11416188 TI - A cellular mechanism for targeting newly synthesized mRNAs to synaptic sites on dendrites. AB - Long-lasting forms of activity-dependent synaptic plasticity involve molecular modifications that require gene expression. Here, we describe a cellular mechanism that mediates the targeting newly synthesized gene transcripts to individual synapses where they are locally translated. The features of this mechanism have been revealed through studies of the intracellular transport and synaptic targeting of the mRNA for a recently identified immediate early gene called activity-regulated cytoskeleton-associated protein Arc. Arc is strongly induced by patterns of synaptic activity that also induce long-term potentiation, and Arc mRNA is then rapidly delivered into dendrites after episodes of neuronal activation. The newly synthesized Arc mRNA localizes selectively at synapses that recently have been activated, and the encoded protein is assembled into the synaptic junctional complex. The dynamics of trafficking of Arc mRNA reveal key features of the mechanism through which synaptic activity can both induce gene expression and target particular mRNA transcripts to the active synapses. PMID- 11416189 TI - Think globally, translate locally: what mitotic spindles and neuronal synapses have in common. AB - Early metazoan development is programmed by maternal mRNAs inherited by the egg at the time of fertilization. These mRNAs are not translated en masse at any one time or at any one place, but instead their expression is regulated both temporally and spatially. Recent evidence has shown that one maternal mRNA, cyclin B1, is concentrated on mitotic spindles in the early Xenopus embryo, where its translation is controlled by CPEB (cytoplasmic polyadenylation element binding protein), a sequence-specific RNA binding protein. Disruption of the spindle-associated translation of this mRNA results in a morphologically abnormal mitotic apparatus and inhibited cell division. Mammalian neurons, particularly in the synapto-dendritic compartment, also contain localized mRNAs such as that encoding alpha-CaMKII. Here, synaptic activation drives local translation, an event that is involved in synaptic plasticity and possibly long-term memory storage. Synaptic translation of alpha-CaMKII mRNA also appears to be controlled by CPEB, which is enriched in the postsynaptic density. Therefore, CPEB controlled local translation may influence such seemingly disparate processes as the cell cycle and synaptic plasticity. PMID- 11416190 TI - Vasopressin mRNA localization in nerve cells: characterization of cis-acting elements and trans-acting factors. AB - mRNA localization is a complex pathway. Besides mRNA sorting per se, this process includes aspects of regulated translation. It requires protein factors that interact with defined sequences (or sequence motifs) of the transcript, and the protein/RNA complexes are finally guided along the cytoskeleton to their ultimate destinations. The mRNA encoding the vasopressin (VP) precursor protein is localized to the nerve cell processes in vivo and in primary cultured nerve cells. Sorting of VP transcripts to dendrites is mediated by the last 395 nucleotides of the mRNA, the dendritic localizer sequence, and it depends on intact microtubules. In vitro interaction studies with cytosolic extracts demonstrated specific binding of a protein, enriched in nerve cell tissues, to the radiolabeled dendritic localizer sequence probe. Biochemical purification revealed that this protein is the multifunctional poly(A)-binding protein (PABP). It is well known for its ability to bind with high affinity to poly(A) tails of mRNAs, prerequisite for mRNA stabilization and stimulation of translational initiation, respectively. With lower affinities, PABP can also associate with non poly(A) sequences. The physiological consequences of these PABP/RNA interactions are far from clear but may include functions such as translational silencing. Presumably, the translational state of mRNAs subject to dendritic sorting is influenced by external stimuli. PABP thus could be a component required to regulate local synthesis of the VP precursor and possibly of other proteins. PMID- 11416191 TI - Local translation of classes of mRNAs that are targeted to neuronal dendrites. AB - The functioning of the neuronal dendrite results from a variety of biological processes including mRNA transport to and protein translation in the dendrite. The complexity of the mRNA population in dendrites suggests that specific biological processes are modulated through the regulation of dendritic biology. There are various classes of mRNAs in dendrites whose translation modulates the ability of the dendrite to receive and integrate presynaptic information. Among these mRNAs are those encoding selective transcription factors that function in the neuronal soma and ionotropic glutamate receptors that function on the neuronal membrane. Conclusive evidence that these mRNAs can be translated is reviewed, and identification of the endogenous sites of translation in living dendrites is presented. These data, as well as those described in the other articles resulting from this colloquium, highlight the complexity of dendritic molecular biology and the exquisitely selective and sensitive modulatory role played by the dendrite in facilitating intracellular and intercellular communication. PMID- 11416192 TI - Cytoskeletal microdifferentiation: a mechanism for organizing morphological plasticity in dendrites. AB - Experimental evidence suggests that microfilaments and microtubules play contrasting roles in regulating the balance between motility and stability in neuronal structures. Actin-containing microfilaments are associated with structural plasticity, both during development when their dynamic activity drives the exploratory activity of growth cones and after circuit formation when the actin-rich dendritic spines of excitatory synapses retain a capacity for rapid changes in morphology. By contrast, microtubules predominate in axonal and dendritic processes, which appear to be morphologically relatively more stable. To compare the cytoplasmic distributions and dynamics of microfilaments and microtubules we made time-lapse recordings of actin or the microtubule-associated protein 2 tagged with green fluorescent protein in neurons growing in dispersed culture or in tissue slices from transgenic mice. The results complement existing evidence indicating that the high concentrations of actin present in dendritic spines is a specialization for morphological plasticity. By contrast, microtubule associated protein 2 is limited to the shafts of dendrites where time-lapse recordings show little evidence for dynamic activity. A parallel exists between the partitioning of microfilaments and microtubules in motile and stable domains of growing processes during development and between dendrite shafts and spines at excitatory synapses in established neuronal circuits. These data thus suggest a mechanism, conserved through development and adulthood, in which the differential dynamics of actin and microtubules determine the plasticity of neuronal structures. PMID- 11416193 TI - Tracking the estrogen receptor in neurons: implications for estrogen-induced synapse formation. AB - Estrogens (E) and progestins regulate synaptogenesis in the CA1 region of the dorsal hippocampus during the estrous cycle of the female rat, and the functional consequences include changes in neurotransmission and memory. Synapse formation has been demonstrated by using the Golgi technique, dye filling of cells, electron microscopy, and radioimmunocytochemistry. N-methyl-d-aspartate (NMDA) receptor activation is required, and inhibitory interneurons play a pivotal role as they express nuclear estrogen receptor alpha (ERalpha) and show E-induced decreases of GABAergic activity. Although global decreases in inhibitory tone may be important, a more local role for E in CA1 neurons seems likely. The rat hippocampus expresses both ERalpha and ERbeta mRNA. At the light microscopic level, autoradiography shows cell nuclear [3H]estrogen and [125I]estrogen uptake according to a distribution that primarily reflects the localization of ERalpha immunoreactive interneurons in the hippocampus. However, recent ultrastructural studies have revealed extranuclear ERalpha immunoreactivity (IR) within select dendritic spines on hippocampal principal cells, axon terminals, and glial processes, localizations that would not be detectable by using standard light microscopic methods. Based on recent studies showing that both types of ER are expressed in a form that activates second messenger systems, these findings support a testable model in which local, non-genomic regulation by estrogen participates along with genomic actions of estrogens in the regulation of synapse formation. PMID- 11416194 TI - Synaptic regulation of protein synthesis and the fragile X protein. AB - Protein synthesis occurs in neuronal dendrites, often near synapses. Polyribosomal aggregates often appear in dendritic spines, particularly during development. Polyribosomal aggregates in spines increase during experience dependent synaptogenesis, e.g., in rats in a complex environment. Some protein synthesis appears to be regulated directly by synaptic activity. We use "synaptoneurosomes," a preparation highly enriched in pinched-off, resealed presynaptic processes attached to resealed postsynaptic processes that retain normal functions of neurotransmitter release, receptor activation, and various postsynaptic responses including signaling pathways and protein synthesis. We have found that, when synaptoneurosomes are stimulated with glutamate or group I metabotropic glutamate receptor agonists such as dihydroxyphenylglycine, mRNA is rapidly taken up into polyribosomal aggregates, and labeled methionine is incorporated into protein. One of the proteins synthesized is FMRP, the protein that is reduced or absent in fragile X mental retardation syndrome. FMRP has three RNA-binding domains and reportedly binds to a significant number of mRNAs. We have found that dihydroxyphenylglycine-activated protein synthesis in synaptoneurosomes is dramatically reduced in a knockout mouse model of fragile X syndrome, which cannot produce full-length FMRP, suggesting that FMRP is involved in or required for this process. Studies of autopsy samples from patients with fragile X syndrome have indicated that dendritic spines may fail to assume a normal mature size and shape and that there are more spines per unit dendrite length in the patient samples. Similar findings on spine size and shape have come from studies of the knockout mouse. Study of the development of the somatosensory cortical region containing the barrel-like cell arrangements that process whisker information suggests that normal dendritic regression is impaired in the knockout mouse. This finding suggests that FMRP may be required for the normal processes of maturation and elimination to occur in cerebral cortical development. PMID- 11416195 TI - Titration of chaos with added noise. AB - Deterministic chaos has been implicated in numerous natural and man-made complex phenomena ranging from quantum to astronomical scales and in disciplines as diverse as meteorology, physiology, ecology, and economics. However, the lack of a definitive test of chaos vs. random noise in experimental time series has led to considerable controversy in many fields. Here we propose a numerical titration procedure as a simple "litmus test" for highly sensitive, specific, and robust detection of chaos in short noisy data without the need for intensive surrogate data testing. We show that the controlled addition of white or colored noise to a signal with a preexisting noise floor results in a titration index that: (i) faithfully tracks the onset of deterministic chaos in all standard bifurcation routes to chaos; and (ii) gives a relative measure of chaos intensity. Such reliable detection and quantification of chaos under severe conditions of relatively low signal-to-noise ratio is of great interest, as it may open potential practical ways of identifying, forecasting, and controlling complex behaviors in a wide variety of physical, biomedical, and socioeconomic systems. PMID- 11416196 TI - Indium-cadmium-oxide films having exceptional electrical conductivity and optical transparency: clues for optimizing transparent conductors. AB - Materials with high electrical conductivity and optical transparency are needed for future flat panel display, solar energy, and other opto-electronic technologies. In(x)Cd(1-x)O films having a simple cubic microstructure have been grown on amorphous glass substrates by a straightforward chemical vapor deposition process. The x = 0.05 film conductivity of 17,000 S/cm, carrier mobility of 70 cm2/Vs, and visible region optical transparency window considerably exceed the corresponding parameters for commercial indium-tin oxide. Ab initio electronic structure calculations reveal small conduction electron effective masses, a dramatic shift of the CdO band gap with doping, and a conduction band hybridization gap caused by extensive Cd 5s + In 5s mixing. PMID- 11416197 TI - The effect of cultivation on the size, shape, and persistence of disease patches in fields. AB - Epidemics of soil-borne plant disease are characterized by patchiness because of restricted dispersal of inoculum. The density of inoculum within disease patches depends on a sequence comprising local amplification during the parasitic phase followed by dispersal of inoculum by cultivation during the intercrop period. The mechanisms that control size, shape, and persistence have received very little rigorous attention in epidemiological theory. Here we derive a model for dispersal of inoculum in soil by cultivation that takes account into the discrete stochastic nature of the system in time and space. Two parameters, probability of movement and mean dispersal distance, characterize lateral dispersal of inoculum by cultivation. The dispersal parameters are used in combination with the characteristic area and dimensions of host plants to identify criteria that control the shape and size of disease patches. We derive a critical value for the probability of movement for the formation of cross-shaped patches and show that this is independent of the amount of inoculum. We examine the interaction between local amplification of inoculum by parasitic activity and subsequent dilution by dispersal and identify criteria whereby asymptomatic patches may persist as inoculum falls below a threshold necessary for symptoms to appear in the subsequent crop. The model is motivated by the spread of rhizomania, an economically important soil-borne disease of sugar beet. However, the results have broad applicability to a very wide range of diseases that survive as discrete units of inoculum. The application of the model to patch dynamics of weed seeds and local introductions of genetically modified seeds is also discussed. PMID- 11416198 TI - Crystal structure of the regulatory subunit H of the V-type ATPase of Saccharomyces cerevisiae. AB - In contrast to the F-type ATPases, which use a proton gradient to generate ATP, the V-type enzymes use ATP to actively transport protons into organelles and extracellular compartments. We describe here the structure of the H-subunit (also called Vma13p) of the yeast enzyme. This is the first structure of any component of a V-type ATPase. The H-subunit is not required for assembly but plays an essential regulatory role. Despite the lack of any apparent sequence homology the structure contains five motifs similar to the so-called HEAT or armadillo repeats seen in the importins. A groove, which is occupied in the importins by the peptide that targets proteins for import into the nucleus, is occupied here by the 10 amino-terminal residues of subunit H itself. The structural similarity suggests how subunit H may interact with the ATPase itself or with other proteins. A cleft between the amino- and carboxyl-terminal domains also suggests another possible site of interaction with other factors. PMID- 11416199 TI - Nitration of succinyl-CoA:3-oxoacid CoA-transferase in rats after endotoxin administration. AB - The tyrosine nitration of proteins has been observed in diverse inflammatory conditions and has been linked to the presence of reactive nitrogen species. From many in vitro experiments, it is apparent that tyrosine nitration may alter the function of proteins. A limited number of experiments under in vivo conditions also demonstrate that protein nitration is associated with altered cellular processes. To understand the association of protein nitration with the pathogenic mechanism of the disease, it is essential to identify specific protein targets of nitration with in vivo or intact tissue models. Using anti-nitrotyrosine antibodies, we demonstrated the accumulation of nitrotyrosine in a 52-kDa protein in rat kidney after lipopolysaccharide treatment. The 52-kDa protein was purified and identified with partial sequence as succinyl-CoA:3-oxoacid CoA-transferase (SCOT; EC ). Western blot analysis revealed that the nitration of this mitochondrial enzyme increased in the kidneys and hearts of lipopolysaccharide treated rats, whereas its catalytic activity decreased. These data suggest that tyrosine nitration may be a mechanism for the inhibition of SCOT activity in inflammatory conditions. SCOT is a key enzyme for ketone body utilization. Thus, tyrosine nitration of the enzyme with sepsis or inflammation may explain the altered metabolism of ketone bodies present in these disorders. PMID- 11416200 TI - Electrospray ionization mass spectrometry analysis of changes in phospholipids in RBL-2H3 mastocytoma cells during degranulation. AB - Biological membranes contain an extraordinary diversity of lipids. Phospholipids function as major structural elements of cellular membranes, and analysis of changes in the highly heterogeneous mixtures of lipids found in eukaryotic cells is central to understanding the complex functions in which lipids participate. Phospholipase-catalyzed hydrolysis of phospholipids often follows cell surface receptor activation. Recently, we demonstrated that granule fusion is initiated by addition of exogenous, nonmammalian phospholipases to permeabilized mast cells. To pursue this finding, we use positive and negative mode Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) to measure changes in the glycerophospholipid composition of total lipid extracts of intact and permeabilized RBL-2H3 (mucosal mast cell line) cells. The low energy of the electrospray ionization results in efficient production of molecular ions of phospholipids uncomplicated by further fragmentation, and changes were observed that eluded conventional detection methods. From these analyses we have spectrally resolved more than 130 glycerophospholipids and determined changes initiated by introduction of exogenous phospholipase C, phospholipase D, or phospholipase A2. These exogenous phospholipases have a preference for phosphatidylcholine with long polyunsaturated alkyl chains as substrates and, when added to permeabilized mast cells, produce multiple species of mono- and polyunsaturated diacylglycerols, phosphatidic acids, and lysophosphatidylcholines, respectively. The patterns of changes of these lipids provide an extraordinarily rich source of data for evaluating the effects of specific lipid species generated during cellular processes, such as exocytosis. PMID- 11416201 TI - Tolerance of human MSH2+/- lymphoblastoid cells to the methylating agent temozolomide. AB - Members of hereditary nonpolyposis colon cancer (HNPCC) families harboring heterozygous germline mutations in the DNA mismatch repair genes hMSH2 or hMLH1 present with tumors generally two to three decades earlier than individuals with nonfamilial sporadic colon cancer. We searched for phenotypic features that might predispose heterozygous cells from HNPCC kindreds to malignant transformation. hMSH2(+/-) lymphoblastoid cell lines were found to be on average about 4-fold more tolerant than wild-type cells to killing by the methylating agent temozolomide, a phenotype that is invariably linked with impairment of the mismatch repair system. This finding was associated with an average 2-fold decrease of the steady-state level of hMSH2 protein in hMSH2(+/-) cell lines. In contrast, hMLH1(+/-) heterozygous cells were indistinguishable from normal controls in these assays. Thus, despite the fact that HNPCC families harboring mutations in hMSH2 or hMLH1 cannot be distinguished clinically, the early stages of the carcinogenic process in hMSH2 and hMLH1 mutation carriers may be different. Should hMSH2(+/-) colonocytes and lymphoblasts harbor a similar phenotype, the increased tolerance of the former to DNA-damaging agents present in the human colon may play a key role in the initiation of the carcinogenic process. PMID- 11416202 TI - Nonnucleoside reverse transcriptase inhibitors are chemical enhancers of dimerization of the HIV type 1 reverse transcriptase. AB - Nonnucleoside reverse transcriptase inhibitors (NNRTIs) are allosteric inhibitors of the HIV type 1 (HIV-1) reverse transcriptase (RT). Yeast grown in the presence of many of these drugs exhibited dramatically increased association of the p66 and p51 subunits of the HIV-1 RT as reported by a yeast two-hybrid assay. The enhancement required drug binding by RT; introduction of a drug-resistance mutation into the p66 construct negated the enhancement effect. The drugs could also induce heterodimerization of dimerization defective mutants. Coimmunoprecipitation of RT subunits from yeast lysates confirmed the induction of heterodimer formation by the drugs. In vitro-binding studies indicate that NNRTIs can bind tightly to p66 but not p51 and then mediate subsequent heterodimerization. This study demonstrates an unexpected effect of NNRTIs on the assembly of RT subunits. PMID- 11416203 TI - Complementary intrastrand base pairing during initiation of Herpes simplex virus type 1 DNA replication. AB - The herpes simplex virus type 1 origin of DNA replication, oriS, contains three copies of the recognition sequence for the viral initiator protein, origin binding protein (OBP), arranged in two palindromes. The central box I forms a short palindrome with box III and a long palindrome with box II. Single-stranded oriS adopts a conformation, oriS*, that is tightly bound by OBP. Here we demonstrate that OBP binds to a box III-box I hairpin with a 3' single-stranded tail in oriS*. Mutations designed to destabilize the hairpin abolish the binding of OBP to oriS*. The same mutations also inhibit DNA replication. Second site complementary mutations restore binding of OBP to oriS* as well as the ability of mutated oriS to support DNA replication. OriS* is also an efficient activator of the hydrolysis of ATP by OBP. Sequence analyses show that a box III-box I palindrome is an evolutionarily conserved feature of origins of DNA replication from human, equine, bovine, and gallid alpha herpes viruses. We propose that oriS facilitates initiation of DNA synthesis in two steps and that OBP exhibits exquisite specificity for the different conformations oriS adopts at these stages. Our model suggests that distance-dependent cooperative binding of OBP to boxes I and II in duplex DNA is succeeded by specific recognition of a box III box I hairpin in partially unwound DNA. PMID- 11416204 TI - Nitric oxide partitioning into mitochondrial membranes and the control of respiration at cytochrome c oxidase. AB - An emerging and important site of action for nitric oxide (NO) within cells is the mitochondrial inner membrane, where NO binds to and inhibits members of the electron transport chain, complex III and cytochrome c oxidase. Although it is known that inhibition of cytochrome c oxidase by NO is competitive with O2, the mechanisms that underlie this phenomenon remain unclear, and the impact of both NO and O2 partitioning into biological membranes has not been considered. These properties are particularly interesting because physiological O2 tensions can vary widely, with NO having a greater inhibitory effect at low O2 tensions (<20 microM). In this study, we present evidence for a consumption of NO in mitochondrial membranes in the absence of substrate, in a nonsaturable process that is O2 dependent. This consumption modulates inhibition of cytochrome c oxidase by NO and is enhanced by the addition of exogenous membranes. From these data, it is evident that the partition of NO into mitochondrial membranes has a major impact on the ability of NO to control mitochondrial respiration. The implications of this conclusion are discussed in the context of mitochondrial lipid:protein ratios and the importance of NO as a regulator of respiration in pathophysiology. PMID- 11416205 TI - Mutations within a furin consensus sequence block proteolytic release of ectodysplasin-A and cause X-linked hypohidrotic ectodermal dysplasia. AB - X-linked hypohidrotic ectodermal dysplasia (XLHED) is a heritable disorder of the ED-1 gene disrupting the morphogenesis of ectodermal structures. The ED-1 gene product, ectodysplasin-A (EDA), is a tumor necrosis factor (TNF) family member and is synthesized as a membrane-anchored precursor protein with the TNF core motif located in the C-terminal domain. The stalk region of EDA contains the sequence -Arg-Val-Arg-Arg156-Asn-Lys-Arg159-, representing overlapping consensus cleavage sites (Arg-X-Lys/Arg-Arg( downward arrow)) for the proprotein convertase furin. Missense mutations in four of the five basic residues within this sequence account for approximately 20% of all known XLHED cases, with mutations occurring most frequently at Arg156, which is shared by the two consensus furin sites. These analyses suggest that cleavage at the furin site(s) in the stalk region is required for the EDA-mediated cell-to-cell signaling that regulates the morphogenesis of ectodermal appendages. Here we show that the 50-kDa EDA parent molecule is cleaved at -Arg156Asn-Lys-Arg(159 downward arrow)- to release the soluble C-terminal fragment containing the TNF core domain. This cleavage appears to be catalyzed by furin, as release of the TNF domain was blocked either by expression of the furin inhibitor alpha1-PDX or by expression of EDA in furin deficient LoVo cells. These results demonstrate that mutation of a functional furin cleavage site in a developmental signaling molecule is a basis for human disease (XLHED) and raise the possibility that furin cleavage may regulate the ability of EDA to act as a juxtacrine or paracrine factor. PMID- 11416206 TI - Use of chimeric proteins to investigate the role of transporter associated with antigen processing (TAP) structural domains in peptide binding and translocation. AB - The transporter associated with antigen processing (TAP) comprises two subunits, TAP1 and TAP2, each containing a hydrophobic membrane-spanning region (MSR) and a nucleotide binding domain (NBD). The TAP1/TAP2 complex is required for peptide translocation across the endoplasmic reticulum membrane. To understand the role of each structural unit of the TAP1/TAP2 complex, we generated two chimeras containing TAP1 MSR and TAP2 NBD (T1MT2C) or TAP2 MSR and TAP1 NBD (T2MT1C). We show that TAP1/T2MT1C, TAP2/T1MT2C, and T1MT2C/T2MT1C complexes bind peptide with an affinity comparable to wild-type complexes. By contrast, TAP1/T1MT2C and TAP2/T2MT1C complexes, although observed, are impaired for peptide binding. Thus, the MSRs of both TAP1 and TAP2 are required for binding peptide. However, neither NBD contains unique determinants required for peptide binding. The NBD-switched complexes, T1MT2C/T2MT1C, TAP1/T2MT1C, and TAP2/T1MT2C, all translocate peptides, but with progressively reduced efficiencies relative to the TAP1/TAP2 complex. These results indicate that both nucleotide binding sites are catalytically active and support an alternating catalytic sites model for the TAP transport cycle, similar to that proposed for P-glycoprotein. The enhanced translocation efficiency of TAP1/T2MT1C relative to TAP2/T1MT2C complexes correlates with enhanced binding of the TAP1 NBD-containing constructs to ATP-agarose beads. Preferential ATP interaction with TAP1, if occurring in vivo, might polarize the transport cycle such that ATP binding to TAP1 initiates the cycle. However, our observations that TAP complexes containing two identical TAP NBDs can mediate translocation indicate that distinct properties of the nucleotide binding site per se are not essential for the TAP catalytic cycle. PMID- 11416207 TI - Carbon- and nitrogen-quality signaling to translation are mediated by distinct GATA-type transcription factors. AB - The target of rapamycin (Tor) proteins sense nutrients and control transcription and translation relevant to cell growth. Treating cells with the immunosuppressant rapamycin leads to the intracellular formation of an Fpr1p rapamycin-Tor ternary complex that in turn leads to translational down regulation. A more rapid effect is a rich transcriptional response resembling that when cells are shifted from high- to low-quality carbon or nitrogen sources. This transcriptional response is partly mediated by the nutrient-sensitive transcription factors GLN3 and NIL1 (also named GAT1). Here, we show that these GATA-type transcription factors control transcriptional responses that mediate translation by several means. Four observations highlight upstream roles of GATA type transcription factors in translation. In their absence, processes caused by rapamycin or poor nutrients are diminished: translation repression, eIF4G protein loss, transcriptional down-regulation of proteins involved in translation, and RNA polymerase I/III activity repression. The Tor proteins preferentially use Gln3p or Nil1p to down-regulate translation in response to low-quality nitrogen or carbon, respectively. Functional consideration of the genes regulated by Gln3p or Nil1p reveals the logic of this differential regulation. Besides integrating control of transcription and translation, these transcription factors constitute branches downstream of the multichannel Tor proteins that can be selectively modulated in response to distinct (carbon- and nitrogen-based) nutrient signals from the environment. PMID- 11416208 TI - Protein misfolding and temperature up-shift cause G1 arrest via a common mechanism dependent on heat shock factor in Saccharomycescerevisiae. AB - Accumulation of misfolded proteins in the cell at high temperature may cause entry into a nonproliferating, heat-shocked state. The imino acid analog azetidine 2-carboxylic acid (AZC) is incorporated into cellular protein competitively with proline and can misfold proteins into which it is incorporated. AZC addition to budding yeast cells at concentrations sufficient to inhibit proliferation selectively activates heat shock factor (HSF). We find that AZC treatment fails to cause accumulation of glycogen and trehalose (Msn2/4 dependent processes) or to induce thermotolerance (a protein kinase C-dependent process). However, AZC-arrested cells can accumulate glycogen and trehalose and can acquire thermotolerance in response to a subsequent heat shock. We find that AZC treatment arrests cells in a viable state and that this arrest is reversible. We find that cells at high temperature or cells deficient in the ubiquitin conjugating enzymes Ubc4 and Ubc5 are hypersensitive to AZC-induced proliferation arrest. We find that AZC treatment mimics temperature up-shift in arresting cells in G1 and represses expression of CLN1 and CLN2. Mutants with reduced G1 cyclin Cdc28 activity are hypersensitive to AZC-induced proliferation arrest. Expression of the hyperstable Cln3-2 protein prevents G1 arrest upon AZC treatment and temperature up-shift. Finally, we find that the EXA3-1 mutation, encoding a defective HSF, prevents efficient G1 arrest in response to both temperature up shift and AZC treatment. We conclude that nontoxic levels of misfolded proteins (induced by AZC treatment or by high temperature) selectively activate HSF, which is required for subsequent G1 arrest. PMID- 11416209 TI - Nuclear hormone receptor CHR3 is a critical regulator of all four larval molts of the nematode Caenorhabditis elegans. AB - CHR3 (nhr-23, NF1F4), the homologue of Drosophila DHR3 and mammalian ROR/RZR/RevErbA nuclear hormone receptors, is important for proper epidermal development and molting in the nematode Caenorhabditis elegans. Disruption of CHR3 (nhr-23) function leads to developmental changes, including incomplete molting and a short, fat (dumpy) phenotype. Here, we studied the role of CHR3 during larval development by using expression assays and RNA-mediated interference. We show that the levels of expression of CHR3 (nhr-23) cycle during larval development and reduction of CHR3 function during each intermolt period result in defects at all subsequent molts. Assaying candidate gene expression in populations of animals treated with CHR3 (nhr-23) RNA-mediated interference has identified dpy-7 as a potential gene acting downstream of CHR3. These results define CHR3 as a critical regulator of all C. elegans molts and begin to define the molecular pathway for its function. PMID- 11416210 TI - Corticosterone levels predict survival probabilities of Galapagos marine iguanas during El Nino events. AB - Plasma levels of corticosterone are often used as a measure of "stress" in wild animal populations. However, we lack conclusive evidence that different stress levels reflect different survival probabilities between populations. Galapagos marine iguanas offer an ideal test case because island populations are affected differently by recurring El Nino famine events, and population-level survival can be quantified by counting iguanas locally. We surveyed corticosterone levels in six populations during the 1998 El Nino famine and the 1999 La Nina feast period. Iguanas had higher baseline and handling stress-induced corticosterone concentrations during famine than feast conditions. Corticosterone levels differed between islands and predicted survival through an El Nino period. However, among individuals, baseline corticosterone was only elevated when body condition dropped below a critical threshold. Thus, the population-level corticosterone response was variable but nevertheless predicted overall population health. Our results lend support to the use of corticosterone as a rapid quantitative predictor of survival in wild animal populations. PMID- 11416211 TI - Retroposon analysis of major cetacean lineages: the monophyly of toothed whales and the paraphyly of river dolphins. AB - SINE (short interspersed element) insertion analysis elucidates contentious aspects in the phylogeny of toothed whales and dolphins (Odontoceti), especially river dolphins. Here, we characterize 25 informative SINEs inserted into unique genomic loci during evolution of odontocetes to construct a cladogram, and determine a total of 2.8 kb per taxon of the flanking sequences of these SINE loci to estimate divergence times among lineages. We demonstrate that: (i) Odontocetes are monophyletic; (ii) Ganges River dolphins, beaked whales, and ocean dolphins diverged (in this order) after sperm whales; (iii) three other river dolphin taxa, namely the Amazon, La Plata, and Yangtze river dolphins, form a monophyletic group with Yangtze River dolphins being the most basal; and (iv) the rapid radiation of extant cetacean lineages occurred some 28-33 million years B.P., in strong accord with the fossil record. The combination of SINE and flanking sequence analysis suggests a topology and set of divergence times for odontocete relationships, offering alternative explanations for several long standing problems in cetacean evolution. PMID- 11416212 TI - Genomic analysis of orthologous mouse and human olfactory receptor loci. AB - Olfactory receptor (OR) genes represent approximately 1% of genomic coding sequence in mammals, and these genes are clustered on multiple chromosomes in both the mouse and human genomes. We have taken a comparative genomics approach to identify features that may be involved in the dynamic evolution of this gene family and in the transcriptional control that results in a single OR gene expressed per olfactory neuron. We sequenced approximately 350 kb of the murine P2 OR cluster and used synteny, gene linkage, and phylogenetic analysis to identify and sequence approximately 111 kb of an orthologous cluster in the human genome. In total, 18 mouse and 8 human OR genes were identified, including 7 orthologs that appear to be functional in both species. Noncoding homology is evident between orthologs and generally is confined within the transcriptional unit. We find no evidence for common regulatory features shared among paralogs, and promoter regions generally do not contain strong promoter motifs. We discuss these observations, as well as OR clustering, in the context of evolutionary expansion and transcriptional regulation of OR repertoires. PMID- 11416213 TI - Nucleotide sequences provide evidence of genetic exchange among distantly related lineages of Trypanosoma cruzi. AB - Simple phylogenetic tests were applied to a large data set of nucleotide sequences from two nuclear genes and a region of the mitochondrial genome of Trypanosoma cruzi, the agent of Chagas' disease. Incongruent gene genealogies manifest genetic exchange among distantly related lineages of T. cruzi. Two widely distributed isoenzyme types of T. cruzi are hybrids, their genetic composition being the likely result of genetic exchange between two distantly related lineages. The data show that the reference strain for the T. cruzi genome project (CL Brener) is a hybrid. Well-supported gene genealogies show that mitochondrial and nuclear gene sequences from T. cruzi cluster, respectively, in three or four distinct clades that do not fully correspond to the two previously defined major lineages of T. cruzi. There is clear genetic differentiation among the major groups of sequences, but genetic diversity within each major group is low. We estimate that the major extant lineages of T. cruzi have diverged during the Miocene or early Pliocene (3-16 million years ago). PMID- 11416214 TI - Interlocked feedback loops contribute to the robustness of the Neurospora circadian clock. AB - Interlocked feedback loops may represent a common feature among the regulatory systems controlling circadian rhythms. The Neurospora circadian feedback loops involve white collar-1 (wc-1), wc-2, and frequency (frq) genes. We show that WC-1 and WC-2 proteins activate the transcription of frq gene, whereas FRQ protein plays dual roles: repressing its own transcription, probably by interacting with the WC-1/WC-2 complex, and activating the expression of both WC proteins. Thus, they form two interlocked feedback loops: one negative and one positive. We establish the physiological significance of the interlocked positive feedback loops by showing that the levels of WC-1 and WC-2 determine the robustness and stability of the clock. Our data demonstrate that with WC-1 being the limiting factor in the WC-1/WC-2 complex, the greater the levels of WC-1 and WC-2, the higher the level of the FRQ oscillation and the more robust the overt rhythms. Our data also show that, despite considerable changes in the levels of WC-1, WC 2, and FRQ, the period of the clock has been limited to a small range, suggesting that the interlocked circadian feedback loops are also important for determining the circadian period length of the clock. PMID- 11416215 TI - Screening insertion libraries for mutations in many genes simultaneously using DNA microarrays. AB - We describe a method to screen pools of DNA from multiple transposon lines for insertions in many genes simultaneously. We use thermal asymmetric interlaced PCR, a hemispecific PCR amplification protocol that combines nested, insertion specific primers with degenerate primers, to amplify DNA flanking the transposons. In reconstruction experiments with previously characterized Arabidopsis lines carrying insertions of the maize Dissociation (Ds) transposon, we show that fluorescently labeled, transposon-flanking fragments overlapping ORFs hybridize to cognate expressed sequence tags (ESTs) on a DNA microarray. We further show that insertions can be detected in DNA pools from as many as 100 plants representing different transposon lines and that all of the tested, transposon-disrupted genes whose flanking fragments can be amplified individually also can be detected when amplified from the pool. The ability of a transposon flanking fragment to hybridize declines rapidly with decreasing homology to the spotted DNA fragment, so that only ESTs with >90% homology to the transposon disrupted gene exhibit significant cross-hybridization. Because thermal asymmetric interlaced-PCR fragments tend to be short, use of the present method favors recovery of insertions in and near genes. We apply the technique to screening pools of new Ds lines using cDNA microarrays containing ESTs for approximately 1,000 stress-induced and -repressed Arabidopsis genes. PMID- 11416216 TI - Neutralizing monoclonal antibodies to hepatocyte growth factor/scatter factor (HGF/SF) display antitumor activity in animal models. AB - The hepatocyte growth factor (HGF/SF) receptor, Met, regulates mitogenesis, motility, and morphogenesis in a cell type-dependent fashion. Activation of Met via autocrine, paracrine, or mutational mechanisms can lead to tumorigenesis and metastasis and numerous studies have linked inappropriate expression of this ligand-receptor pair to most types of human solid tumors. To prepare mAbs to human HGF/SF, mice were immunized with native and denatured preparations of the ligand. Recloned mAbs were tested in vitro for blocking activity against scattering and branching morphogenesis. Our results show that no single mAb was capable of neutralizing the in vitro activity of HGF/SF, and that the ligand possesses a minimum of three epitopes that must be blocked to prevent Met tyrosine kinase activation. In vivo, the neutralizing mAb combination inhibited s.c. growth in athymic nu/nu mice of tumors dependent on an autocrine Met-HGF/SF loop. Importantly, growth of human glioblastoma multiforme xenografts expressing Met and HGF/SF were markedly reduced in the presence of HGF/SF-neutralizing mAbs. These results suggest interrupting autocrine and/or paracrine Met-HGF/SF signaling in tumors dependent on this pathway is a possible intervention strategy. PMID- 11416217 TI - Conditional and targeted overexpression of vascular chymase causes hypertension in transgenic mice. AB - We cloned a rat vascular chymase (RVCH) from smooth muscle cells (SMCs) that converts angiotensin I to II and is up-regulated in SMC from spontaneously hypertensive vs. normotensive rats. To determine whether increased activity of RVCH is sufficient to cause hypertension, transgenic mice were generated with targeted conditional expression of RVCH to SMC, with the use of the tetracycline controlled transactivator (tTA). We confirmed conditional expression of RVCH by mRNA, protein, and chymase activity in the absence, but not in the presence, of dietary doxycycline. The systolic blood pressure (mmHg), measured by carotid artery cannulation at 10-12 weeks of age, was higher in tTA+/RVCH+ mice than in nonbinary transgenic littermates (136 +/- 4 vs. 109 +/- 3) (P < 0.05), as were the diastolic and mean pressures. Hypertension was completely reversed by doxycycline, suggesting a causal link with chymase expression. Medial thickening of mesenteric arteries from tTA+/RVCH+ mice vs. littermates (0.82 +/- 0.1 vs. 0.42 +/- 0.02) (P < 0.05) was associated with increased SMC proliferation, as judged by positive immunoreactivity, with the use of an antibody to the proliferating cell nuclear antigen. These structural changes were prevented by doxycycline. Perfusion myography of mesenteric arteries from tTA+/RVCH+ mice also revealed increased vasoconstriction in response to phenylephrine and impaired metacholine-induced vasodilatation when compared with littermate controls or with the doxycyline-treated group. Our studies suggest that up-regulation of this vascular chymase is sufficient to cause a hypertensive arteriopathy, and that RVCH may be a candidate gene and a therapeutic target in patients with high blood pressure. PMID- 11416218 TI - Gamma -secretase inhibitors repress thymocyte development. AB - A major therapeutic target in the search for a cure to the devastating Alzheimer's disease is gamma-secretase. This activity resides in a multiprotein enzyme complex responsible for the generation of Abeta42 peptides, precipitates of which are thought to cause the disease. Gamma-secretase is also a critical component of the Notch signal transduction pathway; Notch signals regulate development and differentiation of adult self-renewing cells. This has led to the hypothesis that therapeutic inhibition of gamma-secretase may interfere with Notch-related processes in adults, most alarmingly in hematopoiesis. Here, we show that application of gamma-secretase inhibitors to fetal thymus organ cultures interferes with T cell development in a manner consistent with loss or reduction of Notch1 function. Progression from an immature CD4-/CD8- state to an intermediate CD4+/CD8+ double-positive state was repressed. Furthermore, treatment beginning later at the double-positive stage specifically inhibited CD8+ single-positive maturation but did not affect CD4+ single-positive cells. These results demonstrate that pharmacological gamma-secretase inhibition recapitulates Notch1 loss in a vertebrate tissue and present a system in which rapid evaluation of gamma-secretase-targeted pharmaceuticals for their ability to inhibit Notch activity can be performed in a relevant context. PMID- 11416219 TI - CML66, a broadly immunogenic tumor antigen, elicits a humoral immune response associated with remission of chronic myelogenous leukemia. AB - This report describes a tumor-associated antigen, termed CML66, initially cloned from a chronic myelogenous leukemia (CML) cDNA expression library. CML66 encodes a 583-aa protein with a molecular mass of 66 kDa and no significant homology to other known genes. CML66 gene is localized to human chromosome 8q23, but the function of this gene is unknown. CML66 is expressed in leukemias and a variety of solid tumor cell lines. When examined by Northern blot, expression in normal tissues was restricted to testis and heart, and no expression was found in hematopoietic tissues. When examined by quantitative reverse transcription-PCR, expression in CML cells was 1.5-fold higher than in normal peripheral blood mononuclear cells. The presence of CML66-specific antibody in patient serum was confirmed by Western blot and the development of high titer IgG antibody specific for CML66 correlated with immune induced remission of CML in a patient who received infusion of normal donor lymphocytes for treatment of relapse. CML66 antibody also was found in sera from 18-38% of patients with lung cancer, melanoma, and prostate cancer. These findings suggest that CML66 may be immunogenic in a wide variety of malignancies and may be a target for antigen specific immunotherapy. PMID- 11416220 TI - Targeted ablation of the 25-hydroxyvitamin D 1alpha -hydroxylase enzyme: evidence for skeletal, reproductive, and immune dysfunction. AB - The active form of vitamin D, 1alpha,25-dihydroxyvitamin D [1alpha,25(OH)2D], is synthesized from its precursor 25 hydroxyvitamin D [25(OH)D] via the catalytic action of the 25(OH)D-1alpha-hydroxylase [1alpha(OH)ase] enzyme. Many roles in cell growth and differentiation have been attributed to 1,25(OH)2D, including a central role in calcium homeostasis and skeletal metabolism. To investigate the in vivo functions of 1,25(OH)2D and the molecular basis of its actions, we developed a mouse model deficient in 1alpha(OH)ase by targeted ablation of the hormone-binding and heme-binding domains of the 1alpha(OH)ase gene. After weaning, mice developed hypocalcemia, secondary hyperparathyroidism, retarded growth, and the skeletal abnormalities characteristic of rickets. These abnormalities are similar to those described in humans with the genetic disorder vitamin D dependent rickets type I [VDDR-I; also known as pseudovitamin D deficiency rickets (PDDR)]. Altered non-collagenous matrix protein expression and reduced numbers of osteoclasts were also observed in bone. Female mutant mice were infertile and exhibited uterine hypoplasia and absent corpora lutea. Furthermore, histologically enlarged lymph nodes in the vicinity of the thyroid gland and a reduction in CD4- and CD8-positive peripheral T lymphocytes were observed. Alopecia, reported in vitamin D receptor (VDR)-deficient mice and in humans with VDDR-II, was not seen. The findings establish a critical role for the 1alpha(OH)ase enzyme in mineral and skeletal homeostasis as well as in female reproduction and also point to an important role in regulating immune function. PMID- 11416221 TI - Omega 3 but not omega 6 fatty acids inhibit AP-1 activity and cell transformation in JB6 cells. AB - Epidemiological and animal-based investigations have indicated that the development of skin cancer is in part associated with poor dietary practices. Lipid content and subsequently the derived fatty acid composition of the diet are believed to play a major role in the development of tumorigenesis. Omega 3 (omega3) fatty acids, including docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), can effectively reduce the risk of skin cancer whereas omega 6 (omega6) fatty acids such as arachidonic acid (AA) reportedly promote risk. To investigate the effects of fatty acids on tumorigenesis, we performed experiments to examine the effects of the omega3 fatty acids EPA and DHA and of the omega6 fatty acid AA on phorbol 12-tetradecanoate 13-acetate (TPA)-induced or epidermal growth factor (EGF)-induced transcription activator protein 1 (AP-1) transactivation and on the subsequent cellular transformation in a mouse epidermal JB6 cell model. DHA treatment resulted in marked inhibition of TPA- and EGF-induced cell transformation by inhibiting AP-1 transactivation. EPA treatment also inhibited TPA-induced AP-1 transactivation and cell transformation but had no effect on EGF-induced transformation. AA treatment had no effect on either TPA or EGF-induced AP-1 transactivation or transformation, but did abrogate the inhibitory effects of DHA on TPA- or EGF-induced AP-1 transactivation and cell transformation in a dose-dependent manner. The results of this study demonstrate that the inhibitory effects of omega3 fatty acids on tumorigenesis are more significant for DHA than for EPA and are related to an inhibition of AP-1. Similarly, because AA abrogates the beneficial effects of DHA, the dietary ratio of omega6 to omega3 fatty acids may be a significant factor in mediating tumor development. PMID- 11416222 TI - Regulation of the Mycobacterium tuberculosis hypoxic response gene encoding alpha -crystallin. AB - Unlike many pathogens that are overtly toxic to their hosts, the primary virulence determinant of Mycobacterium tuberculosis appears to be its ability to persist for years or decades within humans in a clinically latent state. Since early in the 20th century latency has been linked to hypoxic conditions within the host, but the response of M. tuberculosis to a hypoxic signal remains poorly characterized. The M. tuberculosis alpha-crystallin (acr) gene is powerfully and rapidly induced at reduced oxygen tensions, providing us with a means to identify regulators of the hypoxic response. Using a whole genome microarray, we identified >100 genes whose expression is rapidly altered by defined hypoxic conditions. Numerous genes involved in biosynthesis and aerobic metabolism are repressed, whereas a high proportion of the induced genes have no known function. Among the induced genes is an apparent operon that includes the putative two component response regulator pair Rv3133c/Rv3132c. When we interrupted expression of this operon by targeted disruption of the upstream gene Rv3134c, the hypoxic regulation of acr was eliminated. These results suggest a possible role for Rv3132c/3133c/3134c in mycobacterial latency. PMID- 11416223 TI - Multilocus analysis of extracellular putative virulence proteins made by group A Streptococcus: population genetics, human serologic response, and gene transcription. AB - Species of pathogenic microbes are composed of an array of evolutionarily distinct chromosomal genotypes characterized by diversity in gene content and sequence (allelic variation). The occurrence of substantial genetic diversity has hindered progress in developing a comprehensive understanding of the molecular basis of virulence and new therapeutics such as vaccines. To provide new information that bears on these issues, 11 genes encoding extracellular proteins in the human bacterial pathogen group A Streptococcus identified by analysis of four genomes were studied. Eight of the 11 genes encode proteins with a LPXTG(L) motif that covalently links Gram-positive virulence factors to the bacterial cell surface. Sequence analysis of the 11 genes in 37 geographically and phylogenetically diverse group A Streptococcus strains cultured from patients with different infection types found that recent horizontal gene transfer has contributed substantially to chromosomal diversity. Regions of the inferred proteins likely to interact with the host were identified by molecular population genetic analysis, and Western immunoblot analysis with sera from infected patients confirmed that they were antigenic. Real-time reverse transcriptase-PCR (TaqMan) assays found that transcription of six of the 11 genes was substantially up-regulated in the stationary phase. In addition, transcription of many genes was influenced by the covR and mga trans-acting gene regulatory loci. Multilocus investigation of putative virulence genes by the integrated approach described herein provides an important strategy to aid microbial pathogenesis research and rapidly identify new targets for therapeutics research. PMID- 11416224 TI - Identification of urocortin III, an additional member of the corticotropin releasing factor (CRF) family with high affinity for the CRF2 receptor. AB - The corticotropin-releasing factor (CRF) family of neuropeptides includes the mammalian peptides CRF, urocortin, and urocortin II, as well as piscine urotensin I and frog sauvagine. The mammalian peptides signal through two G protein-coupled receptor types to modulate endocrine, autonomic, and behavioral responses to stress, as well as a range of peripheral (cardiovascular, gastrointestinal, and immune) activities. The three previously known ligands are differentially distributed anatomically and have distinct specificities for the two major receptor types. Here we describe the characterization of an additional CRF related peptide, urocortin III, in the human and mouse. In searching the public human genome databases we found a partial expressed sequence tagged (EST) clone with significant sequence identity to mammalian and fish urocortin-related peptides. By using primers based on the human EST sequence, a full-length human clone was isolated from genomic DNA that encodes a protein that includes a predicted putative 38-aa peptide structurally related to other known family members. With a human probe, we then cloned the mouse ortholog from a genomic library. Human and mouse urocortin III share 90% identity in the 38-aa putative mature peptide. In the peptide coding region, both human and mouse urocortin III are 76% identical to pufferfish urocortin-related peptide and more distantly related to urocortin II, CRF, and urocortin from other mammalian species. Mouse urocortin III mRNA expression is found in areas of the brain including the hypothalamus, amygdala, and brainstem, but is not evident in the cerebellum, pituitary, or cerebral cortex; it is also expressed peripherally in small intestine and skin. Urocortin III is selective for type 2 CRF receptors and thus represents another potential endogenous ligand for these receptors. PMID- 11416225 TI - Identification and characterization of a melanin-concentrating hormone receptor. AB - Melanin-concentrating hormone (MCH), a neuropeptide expressed in central and peripheral nervous systems, plays an important role in the control of feeding behaviors and energy metabolism. An orphan G protein-coupled receptor (SLC 1/GPR24) has recently been identified as a receptor for MCH (MCHR1). We report here the identification and characterization of a G protein-coupled receptor as the MCH receptor subtype 2 (MCHR2). MCHR2 has higher protein sequence homology to MCHR1 than any other G protein-coupled receptor. The expression of MCHR2 has been detected in many regions of the brain. In contrast to MCHR1, which is intronless in the coding region and is located at the chromosomal locus 22q13.3, the MCHR2 gene has multiple exons and is mapped to locus 6q21. MCHR2 is specifically activated by nanomolar concentrations of MCH, binds to MCH with high affinity, and signals through Gq protein. This discovery is important for a full understanding of MCH biology and the development of potential therapeutics for diseases involving MCH, including obesity. PMID- 11416226 TI - Evolution and divergence of sodium channel genes in vertebrates. AB - Invertebrate species possess one or two Na+ channel genes, yet there are 10 in mammals. When did this explosive growth come about during vertebrate evolution? All mammalian Na+ channel genes reside on four chromosomes. It has been suggested that this came about by multiple duplications of an ancestral chromosome with a single Na+ channel gene followed by tandem duplications of Na+ channel genes on some of these chromosomes. Because a large-scale expansion of the vertebrate genome likely occurred before the divergence of teleosts and tetrapods, we tested this hypothesis by cloning Na+ channel genes in a teleost fish. Using an approach designed to clone all of the Na+ channel genes in a genome, we found six Na+ channel genes. Phylogenetic comparisons show that each teleost gene is orthologous to a Na+ channel gene or gene cluster on a different mammalian chromosome, supporting the hypothesis that four Na+ channel genes were present in the ancestors of teleosts and tetrapods. Further duplications occurred independently in the teleost and tetrapod lineages, with a greater number of duplications in tetrapods. This pattern has implications for the evolution of function and specialization of Na+ channel genes in vertebrates. Sodium channel genes also are linked to homeobox (Hox) gene clusters in mammals. Using our phylogeny of Na+ channel genes to independently test between two models of Hox gene evolution, we support the hypothesis that Hox gene clusters evolved as (AB) (CD) rather than [D[A(BC)]]. PMID- 11416227 TI - Antillatoxin is a marine cyanobacterial toxin that potently activates voltage gated sodium channels. AB - Antillatoxin (ATX) is a lipopeptide derived from the pantropical marine cyanobacterium Lyngbya majuscula. ATX is neurotoxic in primary cultures of rat cerebellar granule cells, and this neuronal death is prevented by either N-methyl d-aspartate (NMDA) receptor antagonists or tetrodotoxin. To further explore the potential interaction of ATX with voltage-gated sodium channels, we assessed the influence of tetrodotoxin on ATX-induced Ca2+ influx in cerebellar granule cells. The rapid increase in intracellular Ca2+ produced by ATX (100 nM) was antagonized in a concentration-dependent manner by tetrodotoxin. Additional, more direct, evidence for an interaction with voltage-gated sodium channels was derived from the ATX-induced allosteric enhancement of [3H]batrachotoxin binding to neurotoxin site 2 of the alpha subunit of the sodium channel. ATX, moreover, produced a strong synergistic stimulation of [3H]batrachotoxin binding in combination with brevetoxin, which is a ligand for neurotoxin site 5 on the voltage-gated sodium channel. Positive allosteric interactions were not observed between ATX and either alpha-scorpion toxin or the pyrethroid deltamethrin. That ATX interaction with voltage-gated sodium channels produces a gain of function was demonstrated by the concentration-dependent and tetrodotoxin-sensitive stimulation of 22Na+ influx in cerebellar granule cells exposed to ATX. Together these results demonstrate that the lipopeptide ATX is an activator of voltage-gated sodium channels. The neurotoxic actions of ATX therefore resemble those of brevetoxins that produce neural insult through depolarization-evoked Na+ load, glutamate release, relief of Mg2+ block of NMDA receptors, and Ca2+ influx. PMID- 11416231 TI - Effect of human chorionic gonadotropin derivatives on leydig cell function. AB - BACKGROUND: Several human chorionic gonadotropin (hCG) derivatives have been detected in healthy human subjects, indicating that they may play a role in cell function. These hCG derivatives include deglycosylated hCG, proteolytic digestion products of hCG and free alpha and beta subunits of the hormone. It is well documented that testicular Leydig cells are responsive to luteinising hormone (LH) or its analogue hCG. These hormones have high affinity for LH/hCG receptors on the plasma membrane. METHODS: We designed functional and binding studies to compare the effects of native hCG and several hCG derivatives on a rat Leydig cell system. The molecular weight of the hCG derivatives was determined by SDS PAGE and the binding affinity to LH/hCG receptors was measured by a radioligand assay. In addition, their ability to produce testosterone, cyclic AMP and arachidonic acid release was also studied. RESULTS: These hCG derivatives, with the exception of the free beta subunit, were able to bind to LH/hCG plasma membrane receptors with different affinities than that of native hCG. In addition, hCG derivatives did not increase intracellular cAMP levels or arachidonic acid release. However, they did increase testosterone production. CONCLUSION: Taken together, the results of this study lead us to suggest that these hCG derivatives may regulate the action of the native hormone in Leydig cells and are, thus, molecules of physiological relevance. PMID- 11416232 TI - Height, bone mineral density and bone markers in congenital adrenal hyperplasia. AB - AIM: To evaluate height, bone growth, areal bone mineral density (aBMD), volumetric bone mineral density (vBMD) and markers of bone turnover in a group of patients affected by congenital adrenal hyperplasia (CAH). PATIENTS: There were 50 patients (23 males, 27 females), aged 1-28 years, affected by CAH due to 21 hydroxylase deficiency: 27 with the salt-wasting (SW); 14 with the simple virilizing (SV), and 9 with the nonclassical (NC) forms. METHODS: Bone morphometry was evaluated with the metacarpal index (MI) and lumbar aBMD and vBMD (L2-L4) by dual energy X-ray absorptiometry. Serum osteocalcin was used as a marker of bone formation, while urinary cross-linked N-telopeptides of type-I collagen and free deoxypyridinoline levels were evaluated as indexes of bone resorption. RESULTS: The height standard deviation score (SDS) was -0.41 +/- 1.4 in SW patients, -0.01 +/- 1.9 in SV patients, and -0.01 +/- 2.3 in NC patients. There was no significant difference among groups and against zero. The MI SDS was also not different between groups and against zero. aBMD was significantly lower in the pubertal patients compared with normal values, but only when patients with the SW and SV forms were considered together (p < 0.05). vBMD was significantly reduced in all patients with the classical form. Bone markers were not different in patients and controls. CONCLUSION: Our study shows that normal height can be attained in CAH patients; however, an impairment in bone growth and mineralization may be found in adolescents and young adults affected by the classical form. PMID- 11416233 TI - Relationship between the serum level of leptin and life-style habits in Japanese men. AB - AIMS/METHODS: To investigate whether smoking affects the serum level of leptin, 708 male workers aged 25-65 years old were cross-sectionally surveyed. RESULTS: Multiple regression analysis indicated that among the various parameters examined, the level of leptin was positively associated with the body mass index and the levels of insulin, total cholesterol and uric acid, and was inversely associated with physical activity and the level of creatinine. The partial correlation coefficient of leptin was highest against the body mass index (r = 0.40), followed by insulin (0.29) and physical activity (-0.14), after adjustment for other leptin-related variables. However, no association was observed between the level of leptin and smoking (0.05), alcohol consumption (0.09) or age (0.09). CONCLUSIONS: The findings suggest that among life-style habits, physical activity, but not smoking or alcohol consumption, significantly affects the serum level of leptin in Japanese men. PMID- 11416234 TI - Time relationship between circadian variation of serum levels of leptin, insulin and cortisol in healthy subjects. AB - BACKGROUND: Leptin is involved in the regulation of eating behavior. Its serum levels are determined by fat mass but a diurnal rhythm is also described. It is not clear whether leptin levels are also controlled in vivo by hormonal stimuli, like insulin or cortisol. METHODS AND RESULTS: This possible temporal relation was investigated by serial measurements during 24 h (group A) and 46 h (group B) in 15 healthy volunteers and another 10 subjects (group C) while fasting for 72 h. Maximal leptin levels were observed at 4:00 a.m. and 4:00 p.m. in subjects on a normal diet. During 24 h starvation (group B), there was a 40% decrease of mean leptin concentration when compared to baseline values. In group C, the leptin concentration under starvation dropped to 25% of basal levels after 72 h. Pooled data from group A and the nonfasting data from group B showed an insulin increase preceding leptin increase by 6 h (r = 0.405, p < 0.0001), while cortisol decreased 4 h (r = 0.361, p < 0.001) after leptin decrease. CONCLUSION: Starvation results in a fall of circulating leptin, ending leptin rhythmicity. Food intake is causally involved in the fluctuation of leptin levels in serum. Presumably this effect is mediated by insulin, while cortisol does not seem to affect leptin release directly in vivo. PMID- 11416235 TI - Influence of sex and age at onset on autoantibodies against insulin, GAD65 and IA2 in recent onset type 1 diabetic patients. AB - METHODS: Autoantibodies against insulin (IAA), glutamic acid decarboxylase (GADA) and tyrosine phosphatase IA2 (IA2A) were measured in sera from 448 recent onset patients with type 1 diabetes mellitus (DM) subdivided according to sex (194 female and 254 male) and age at onset (134 patients diagnosed before 10 years, 187 between 10 and 20 years, 66 between 20 and 30 years and 61 over 30 years. RESULTS: Autoantibodies were more frequent in female DM patients (93.8 vs. 86.6%, p = 0.013) due to an increased prevalence of both GADA (86.1 vs. 70.1%) and IA2A (59.3 vs. 49.2%), with GADA levels also significantly higher in women (0.24 vs. 0.18 U, p = 0.0003). When age groups were compared, there was a reduction in prevalence in patients over 20 years for both IAA (70% for patients diagnosed under 20 and 36% for older patients) and IA2A (65 and 25%, respectively). These differences also affected IAA levels, with the highest antibody titres in the youngest group (1,214.1 nU/ml in children under 10 compared to 546.9, 345.6 and 341.1 nU/ml in the subsequent groups; p < 10(-4)). GADA prevalence did not differ significantly between age groups but, nevertheless, autoantibody levels were highest among the oldest type 1 DM patients (0.327 U compared to 0.216, 0.197 and 0.176 U in the decreasing age groups; p < 10(-4)). CONCLUSION: There are sex- and age-related differences affecting the presence and/or titres of beta cell autoantibodies. We speculate that these differences could reflect the severity and specificity of the autoimmune attack against the endocrine pancreas and might influence the rate of progression to type 1 DM or the risk of developing other autoimmune diseases. PMID- 11416236 TI - Insulin secretion and acetylcholinesterase activity in monosodium l-glutamate induced obese mice. AB - OBJECTIVE: Pancreatic islets isolated from mice treated neonatally with monosodium L-glutamate (MSG) were used to study insulin secretion. MATERIALS AND METHODS: Total acetylcholinesterase (AchE) activity of tissue extract was measured as a cholinergic activity marker. Obesity recorded in 90-day-old MSG mice (OM) by Lee index reached 366.40 +/- 1.70, compared to control mice (CM) 324.40 +/- 1.10 (p < 0.0001). Glucose 5.6 mM induced insulin secretion of 36 +/- 5 pg/15 min from islets of CM and 86 +/- 13 from OM (p < 0.001). When glucose was raised to 16.7 mM, islets from OM secreted 1,271 +/- 215 and 1,017 +/- 112 pg/30 min to CM. AchE activity of pancreas from OM was 0.64 +/- 0.02 nmol of substrate hydrolyzed/min/mg of tissue and 0.52 +/- 0.01 to CM (p < 0.0001). Liver of obese animals also presented increase of AchE activity. RESULTS: These indicate that OM insulin oversecretion in low glucose may be attributed, at least in part, to an enhancement of parasympathetic tonus. PMID- 11416237 TI - Increased bone mineral density and serum leptin in non-obese girls with precocious pubarche: relation to low birthweight and hyperinsulinism. AB - BACKGROUND: Hyperinsulinism and hyperandrogenism have the capacity to increase bone mineral density (BMD) and serum leptin, independently of body fat mass. We therefore assessed lumbar BMD and serum leptin in girls with the sequence of a low birthweight and precocious pubarche (PP) in childhood, in whom hyperinsulinism and hyperandrogenism have been described. METHODS: Fifty-two non obese PP girls were studied (age range 6.9-14.9 years). Serum leptin was also measured in 42 control girls, matched for age, body mass index and pubertal stage. RESULTS: BMD SDS, measured by dual-energy X-ray absorptiometry, was elevated in PP girls compared to the population reference (0.39 +/- 0.18 SDS; p = 0.03) and bone age, assessed from hand radiographs, was significantly advanced compared to chronological age (1.2 +/- 0.1 years; p < 0.0005). CONCLUSION: Compared to control girls, PP girls had higher leptin levels for degree of body mass index (PP girls: 9.4 +/- 0.6 ng/ml; controls: 7.8 +/- 0.6 ng/ml; p = 0.01). In PP girls, serum leptin was inversely related to birthweight (r = -0.32, p = 0.01) and positively related to free androgen index (FAI) (r = 0.71, p < 0.0005). BMD SDS was also inversely related to birthweight (r = -0.26, p < 0.05) and positively related to serum leptin (r = 0.42, p < 0.05), FAI (r = 0.45, p < 0.05) and mean serum insulin during oral glucose tolerance testing (MSI) (r = 0.59, p < 0.0005). In multiple regression, MSI was the strongest determinant of BMD SDS (beta = 0.50, p = 0.002). In conclusion, elevated BMD and serum leptin in non obese PP girls were related to degrees of low birthweight, hyperinsulinism and hyperandrogenism. The characteristic hyperinsulinism of PP girls is proposed to be the key variable in this constellation. PMID- 11416238 TI - Significant gene expression of insulin-like growth factor II and proliferating cell nuclear antigen in a rapidly growing recurrent pituitary acth-secreting adenoma. AB - BACKGROUND: We quantified the expression of various growth-related factors in an adrenocorticotropic hormone (ACTH)-secreting adenoma that had recurred very rapidly as invasive macroadenoma. METHODS/RESULTS: A 43-year-old woman underwent successful transsphenoidal surgery for Cushing's disease. Seven years later, she was admitted to our ward for further endocrine examinations. In spite of a very high plasma ACTH level, the serum cortisol level was normal. Discrepancies between ACTH and cortisol levels were detected on the basis of diurnal rhythms, dexamethasone suppression tests, and corticotropin-releasing hormone test. The patient showed no clinical features of Cushing's disease. Magnetic resonance imaging of the pituitary showed an almost empty sella, and no microadenoma was found. These results, along with those of Sephadex column gel filtration and high performance liquid chromatography of plasma-immunoreactive ACTH, suggested that the patient's residual corticotrophs secreted biologically inactive ACTH. Two years later, the patient suddenly developed diplopia and right abducens nerve palsy. She was slightly moonfaced and centrally obese. Her plasma ACTH and serum and urinary free cortisol levels were elevated, although discrepancies between ACTH and cortisol still existed. Magnetic resonance imaging revealed a large pituitary mass with suprasellar and cavernous sinus extensions. The tumor was excised, and the proopiomelanocortin gene and the expression of growth-related factors were analyzed. No mutations were found in the ACTH-coding region of the proopiomelanocortin gene. A significant expression of insulin-like growth factor II and proliferating cell nuclear antigen mRNAs was demonstrated. A high MIB-1 antibody labeling index was also detected in the adenoma tissue, suggesting high Ki-67 expression. CONCLUSION: These growth- and proliferation-related factors might be involved in the rapid growth and aggressiveness of this patient's pituitary adenoma. PMID- 11416239 TI - Effects of 22K or 20K human growth hormone on lipolysis, leptin production in adipocytes in the presence and absence of human growth hormone binding protein. AB - OBJECTIVE AND METHOD: We studied the effects of human growth hormone (hGH) on leptin production and lipolysis stimulation in the presence or absence of human growth hormone binding protein (hGHBP) using 3T3- L1-hGHR adipocytes which efficiently express human growth hormone receptor. RESULTS AND CONCLUSION: It was clarified that (1) hGH decreases leptin secretion after hGH-induced lipolysis stimulation, and (2) the reduction of leptin production and lipolysis stimulation by 22K hGH was attenuated with hGHBP, whereas that by 20K hGH, which is a naturally occurring isoform of 22K hGH, was not affected with hGHBP. PMID- 11416240 TI - How and why exercise is impaired in COPD. AB - Published data indicate that exercise in COPD is more often limited by leg effort than breathlessness. This casts some doubt on the classical belief that inability to ventilate limits exercise performance. In fact, symptoms limiting exercise appear to be essentially the same in COPD and in health or congestive heart failure, where exercise is limited by inadequate energy supply to locomotor muscles. In COPD, impaired O2 delivery to locomotor muscles is suggested by: (1) the O2 cost (VO2) of breathing may be approximately 50% of whole body VO2; (2) decreasing the work of breathing improves performance and VO2 of locomotor muscles, and (3) locomotor muscle VO2 is greater when it is the only muscle exercising than during whole body exercise. Excessive expiratory pressures when expiratory flow is limited may lead to decreased venous return and contribute importantly to exercise limitation. PMID- 11416241 TI - The hazards of measuring the drive to breathe. PMID- 11416242 TI - Wheeze and eosinophilic airway inflammation equals asthma? PMID- 11416243 TI - Acute effect of nasal continuous positive air pressure on the ventilatory control of patients with obstructive sleep apnea. AB - BACKGROUND: Sleep fragmentation can decrease the awake ventilatory control. Since patients with obstructive sleep apnea (OSA) patients exhibit sleep fragmentation linked to respiratory events, their ventilatory control could be impaired. However, most of these patients are also obese, which could conversely increase the ventilatory control. The effect of nasal continuous positive airway pressure (CPAP) on the awake ventilatory control in normocapnic OSA patients is unclear. OBJECTIVES: To study the acute effect of nasal CPAP on the awake ventilatory control in normocapnic OSA patients. METHODS: 12 normocapnic OSA patients, with an apnea/hypopnea index (AHI) >15 with moderate obesity (body mass index: 33.5 kg/m2) and normal pulmonary function tests were submitted to two polysomnography studies (diagnostic and for CPAP titration). Before and after 3 consecutive nights of nasal CPAP we analyzed the hypersomnia score and the ventilatory and the mouth occlusion pressure (P.1) responses at rest (breathing room air and a mixture of 8% CO2 + 40% O2). RESULTS: The respiratory drive of OSA patients as evaluated by P.1 was in the range of the controls of our laboratory. After nasal CPAP, a significant decrease in AHI (mean: 51.9-9.4/h) and arousal (mean: 88.7 43/h) occurred, as well as improvement in nocturnal oxyhemoglobin. There was a marginal increase in DeltaV(E)/DeltaP(ET)CO2 (mean: 1.41-1.87 liters/min/ mm Hg, p = 0.09) and a significant rise in P.1/DeltaP(ET)CO2 (mean: 0.29-0.43 cm H2O/mm Hg), a better indicator of ventilatory drive. CONCLUSIONS: Normocapnic OSA patients increased their awake ventilatory drive response to a hypercapnic and hyperoxic mixture with the use of 3 consecutive nights of nasal CPAP. PMID- 11416244 TI - Obstructive sleep apnoea syndrome is common in subjects with chronic bronchitis. Report from the Obstructive Lung Disease in Northern Sweden studies. AB - BACKGROUND: In previous studies we have found that subjects with bronchitis have a higher prevalence of 'snoring as a problem' than respiratory healthy subjects. OBJECTIVES: We aimed to study whether the high prevalence of snoring among subjects with bronchitis also represents a high prevalence of obstructive sleep apnoea (OSA). METHOD: Subjects in three age groups born 1919-1920, 1934-1935 and 1949-1950 had been identified as bronchitic in an earlier study (n = 471) and without respiratory symptoms (n = 108). Of the 91 subjects reporting snoring to be a problem, 70 were invited to participate in the study. Sleep investigation was performed in 52 of these 70 subjects. RESULTS: 'Snoring as a problem', predicted OSA to a similar degree in both bronchitic and respiratory healthy subjects. The estimated prevalence for obstructive sleep apnoea with an apnoea/hypopnoea index (AHI) 10 as the cut-off point and concomitant daytime symptoms such as daytime sleepiness or liability to nodding off during breaks in activity in the daytime, was 5.4% for bronchitic subjects and 2.3% for respiratory healthy subjects. Apnoea in addition to snoring predicted OSA better than did snoring alone. Age correlated significantly with AHI, and OSA was most common in the middle-aged group, 61-62 years old. CONCLUSION: OSA is twice as common in subjects with chronic bronchitis as in subjects free of pulmonary disease or symptoms. PMID- 11416245 TI - A population-based prescription study of asthma drugs during pregnancy: changing the intensity of asthma therapy and perinatal outcomes. AB - BACKGROUND: Among the goals of gestational asthma, therapy is optimisation of pulmonary function. According to the US Food and Drug Administration, no asthma drugs can be considered 'safe' during pregnancy. Fear of adverse fetal effects may thus lead to restrictive use of asthma drugs during pregnancy, and no population-based studies concerning gestational asthma therapy exist. OBJECTIVES: To examine whether asthma drugs or changing intensity of asthma therapy during pregnancy was associated with deviations from expected values of gestational age, birth weight, length at birth, or malformations. METHODS: The Birth Registry was used to identify all 15,756 primiparous women who gave birth in the County of North Jutland between 1991 and 1996. According to the North Jutland Prescription Database, 303 of these women received prescriptions for asthma drugs during pregnancy. Women who did not purchase any prescription drugs during pregnancy constituted the reference group. CONCLUSION: Women who received prescriptions for asthma drugs during pregnancy gave birth to infants with birth weight and length at birth within the expected limits. Reducing intensity of asthma treatment during pregnancy was associated with lower birth weight and length at birth. This may indicate that pregnant women chose to discontinue therapy although their disease severity justifies continuation of treatment. However, analyses did not take into account important clinical variables, and results could also be due to confounding factors or chance. PMID- 11416246 TI - Inhibition of lung natural killer cell activity by smoking: the role of alveolar macrophages. AB - BACKGROUND: It is known that natural killer (NK) cell activity in the lung of smokers (SM) is lower than in non-smokers (NS). However, little is known about the underlying mechanisms. OBJECTIVE: The purpose of this work was to investigate the mechanisms of the inhibition of NK cell activity by alveolar macrophages (AM) in SM. METHODS: Lung effector cells and AM were obtained using bronchoalveolar lavage. The NK cell activity was assayed by 51Cr release method after incubation of 4 and 24 h, using K562 as target cell. AM were added at a concentration of 25% to effector cells. RESULTS: Following 24-hour culture, NK cell activity significantly increased in the NS but not in the SM. Lung NK cell activity was significantly augmented by interleukin-2 in the NS but not in the SM. Addition of AM to the NK cell preparation from SM exerted a significantly greater suppressive effect on autologous blood NK cell activity than in the NS. Indomethacin, catalase or thiourea did not prevent AM-mediated suppression of NK cell activity, in contrast to superoxide dismutase. CONCLUSIONS: These results suggest that the suppression of NK cell activity by AM in SM may be caused by O2- release rather than by prostaglandins, H2O2 or OH release from AM. PMID- 11416247 TI - Pulmonary function in non-insulin-dependent diabetes mellitus. AB - BACKGROUND: In type I diabetes mellitus, lung function has been investigated in several clinical studies, but there are few data concerning pulmonary function abnormalities in patients with non-insulin-dependent diabetes mellitus (NIDDM). OBJECTIVES: The aim of this study was to assess the presence of pulmonary function abnormalities in patients with NIDDM and to verify the possible associations between diabetic renal microangiopathy, retinopathy and diabetes control. METHOD AND PATIENTS: Thirty patients with NIDDM were collected and divided into two similar groups: subjects with retinopathy and/or diabetic glomerulopathy (group 1, n = 15) and patients without any complications (group 2, n = 15). 17 were males and 13 females, aged from 45 to 81 years. They had had diabetes for 3-23 years and were studied at the Division of Internal Medicine, with an outpatient service for diabetic patients. All patients were non-smokers. The presence of diabetic glomerulopathy was determined by measuring the 24-hour protein excretion rate using the nephelometric method. The presence of retinopathy was determined by using ophthalmoscopy. Glycosylated hemoglobin was measured as an indicator of glycemic control. We performed a global spirometry and measured pulmonary diffusion capacity by the single-breath method corrected by alveolar volume. RESULTS: We found a significant reduction in lung diffusion capacity for carbon monoxide (DL(CO)) in the group of patients with other signs of diabetic microangiopathy (p < 0.005) and a significative correlation between DL(CO )and the grade of albuminuria (r = -0.83, p < 0.001). CONCLUSIONS: Pulmonary function abnormalities, in particular a reduction in diffusion capacity, are common in patients with NIDDM and signs of diabetic microangiopathy. A possible explanation is related to an impaired pulmonary microvasculature and alveolar epithelial basal lamina. PMID- 11416248 TI - Evidence of a restrictive spirometric pattern in older thalassemic patients. AB - BACKGROUND: Lung impairment represents one of the complications of thalassemia major whose clinical picture can remain in subclinical form all life long. Few works have been published and the results are contradictory. OBJECTIVES: The aim of this study was to determine the spirometric pattern, the age of onset in thalassemic boys and girls, and to investigate the association with the onset of endocrinological complications. METHODS: We studied 48 patients, divided into three groups according to pubertal stages, in order to better distinguish the periods of life in which different endocrinological complications usually appear. Group A: (n = 14; 8 F, 6 M; age 10.8 +/- 1.7 years): prepubertal patients; group B (n = 21; 10 F, 11 M; age 15.7 +/- 1.1 years): pubertal patients; and group C (n = 13; 9 F, 4 M; age 19.0 +/- 1.4 years): postpubertal patients. Pulmonary function tests (PFTs) and diffusing capacity for carbon monoxide (DCO), corrected for both Hb values (DCO*) and alveolar volume (KCO), were performed 2 days after blood transfusion and were considered pathologic when they fell below 80% of the predicted value. RESULTS: All patients in group A showed normal PFTs, DCO* and KCO values, as well as normal endocrinological assessment. By contrast, all those in group C showed a restrictive spirometric pattern with reduced DCO* (63 +/- 8%), elevated KCO values (120 +/- 14%), a variable degree of hypoxia (PO2 82 +/- 9%), and high serum ferritin levels. Only 2 patients showed a radiological picture of interstitial fibrosis. Furthermore, 9 patients had hypogonadism and 3 hypothyroidism. In group B, only 3 patients showed a restrictive pattern and 1 of them reduced DCO* values, and 2 out of 3 patients had hypogonadism. Considering all patients on the whole, DCO* was negatively correlated with both serum ferritin (r = -0.58; p < 0.05) and age (r = -0.57; p < 0.05). CONCLUSIONS: The presence of a restrictive pattern in 16 older patients associated with both high serum ferritin levels and endocrinological complications in a lot of them was the main feature in this study. Iron overload might be the main factor determining lung impairment, even though a more accurate evaluation is necessary. Possible pathological mechanisms and the role of the genotype are discussed. PMID- 11416249 TI - Central venous and mixed venous oxygen saturation in critically ill patients. AB - BACKGROUND: Although mixed venous O2 saturation (SvO2) accurately indicates the balance of O2 supply/demand and provides an index of tissue oxygenation, the use of a pulmonary artery (PA) catheter is associated with significant costs, risks and complications. Central venous O2 saturation (ScvO2), obtained in a less risky and costly manner, can be an attractive alternative to SvO2. OBJECTIVES: To investigate whether the values of ScvO2 and SvO2 are well correlated and interchangeable in the evaluation of critically ill ICU patients and to create an equation that could estimate SvO2 from ScvO2. METHODS: Sixty-one mechanically ventilated patients were catheterized upon admission and ScvO2 and SvO2 values were simultaneously measured in the lower part of the superior vena cava and PA respectively. RESULTS: SvO2 was 68.6 +/- 1.2% (mean +/- SEM) and ScvO2 was 69.4 +/- 1.1%. The difference is statistically significant (p < 0.03). The correlation coefficient r is 0.945 for the total population, 0.937 and 0.950 in surgical and medical patients, respectively. In 90.2% of patients the difference was <5%. When regression analysis was performed, among 11 models tested, power model [SvO2 = b0(ScvO2)b1] best described the relationship between the two parameters (R2 = 0.917). CONCLUSIONS: ScvO2 and SvO2 are closely related and are interchangeable for the initial evaluation of critically ill patients even if cardiac indices are different. SvO2 can be estimated with great accuracy by ScvO2 in 92% of the patients using a power model. PMID- 11416250 TI - N-omega-nitro-L-arginine methyl ester increases airway responsiveness to serotonin but not to acetylcholine in cats in vivo. AB - BACKGROUND: We previously reported that N(omega)-nitro-L-arginine methyl ester (L NAME) enhances airway responsiveness to inhaled serotonin in cats treated with atropine and propranolol. OBJECTIVE: To further elucidate the role of nitric oxide (NO) in airway responsiveness, we investigated whether L-NAME induces airway hyperresponsiveness to serotonin and acetylcholine (ACh) in animals with intact innervation. METHODS: Cats were anesthetized with pentobarbital sodium (50 mg/kg, i.p.), and mechanically ventilated. To assess airway responsiveness, we measured increase in total pulmonary resistance (RL) produced by delivering serotonin or ACh aerosol to the airway, and determined PC200 (the concentration which caused a 200% increase in RL). RESULTS: The following results were obtained: (1) Airway responsiveness to serotonin was significantly enhanced by the administration of L-NAME (100 mg/kg) in animals treated with atropine and propranolol. (2) Airway responsiveness to serotonin was also significantly enhanced by L- NAME in animals with intact innervation. (3) In contrast, airway responsiveness to ACh was not changed by the addition of L-NAME in cats with intact innervation. CONCLUSION: These results suggest that NO modulates nonspecific airway responsiveness in animals with intact innervation, presumably by a reflex mechanism. PMID- 11416251 TI - Addition of ATP and MgCl2 to the preservation solution attenuates lung reperfusion injury following cold ischemia. AB - BACKGROUND: In lung transplantation, reperfusion injury following cold ischemia is one of the crucial problems for recipients. OBJECTIVE: We evaluated the protective effect of adding a combination of ATP and MgCl2 to the preservation solution against lung reperfusion injury following cold ischemia. METHODS: Using an isolated rat lung perfusion model with fresh rat blood as the perfusate, the rats were divided into five groups (n = 6). In the fresh group, the study lungs were flushed with phosphate-buffered saline (PBS), then immediately reperfused for 120 min. In the control group, the study lungs were flushed with PBS, then cold ischemia was induced for 9 h (4 degrees C), after which reperfusion was performed. In the other three groups, the protocols were the same as for the control group except that ATP and/or MgCl2 were added to the PBS: ATP group (100 microM ATP), MgCl2 group (100 microM MgCl2) and ATP + MgCl2 group (100 microM ATP + 100 microM MgCl2). RESULTS: In the ATP + MgCl2 group, the intrapulmonary shunt fraction, peak airway pressure and wet to dry lung weight ratio were significantly lower than those in the control group. No improvement was observed in the ATP or MgCl2 groups. Histological examination supported these physiological results. In all groups, flush time and lipid peroxide levels in the lungs after cold ischemia did not show any significant differences. CONCLUSION: The addition of ATP and MgCl2 to the preservation solution attenuated reperfusion injury following cold ischemia in rat lungs. PMID- 11416252 TI - Parallel induction of nitric oxide and tetrahydrobiopterin synthesis in alveolar macrophages. AB - BACKGROUND: Nitric oxide (NO) and an essential cofactor for both constitutive and inducible NO synthase (NOS) activity, tetrahydrobiopterin (6R-L-erythro-1',2' dihydroxypropyl-2-amino-4-hydroxy-5,6,7,8-tetrahydropteridine; BH4), are thought to be important modulators of function in normal and inflamed airways. However, the exact pathologic roles of NO and BH4 remain obscure. Even less is known about the effects of cytokines on alveolar macrophages. OBJECTIVE: This study was designed to determine whether NO and BH4 are induced by cytokines in mouse alveolar macrophages and to investigate whether NO synthesis is affected by changes in intracellular BH4 levels in alveolar macrophages. METHODS: We compared the induction by lipopolysaccharide (LPS), interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), and interleukin-2 (IL-2) of NO production and BH4 synthesis in alveolar macrophages. To determine whether NO synthesis is affected by changes in intracellular BH4 levels in alveolar macrophages, we used inhibitors of BH4 biosynthesis. RESULTS: Activation of alveolar macrophages induced parallel increases in NO and intracellular BH4 levels, although induction of the latter appears to be somewhat more sensitive than that of the latter to diverse cytokines. Inducible NO production in alveolar macrophages was blocked by inhibitors of BH4 biosynthesis. IL-2, an important component of the immunomodulatory system, was only a weak activator of alveolar macrophages by itself but potently synergized with IFN-gamma to stimulate the production of both NO and BH4. CONCLUSION: Our results suggest that BH4 synthesis in alveolar macrophages is a potential target for therapeutic intervention in airway inflammatory diseases, such as asthma, cystic fibrosis, and acute bronchial infections whose pathology may be mediated by overproduction of NO. PMID- 11416253 TI - Evaluation of a portable recording device (Somnocheck) for use in patients with suspected obstructive sleep apnoea. AB - BACKGROUND: Portable recording devices for the diagnosis of obstructive sleep apnoea (OSA) should be carefully evaluated before being used on a routine basis in out-patients. OBJECTIVE: This study was designed to evaluate the diagnostic accuracy of the new portable recording device, Somnocheck (SC). METHODS: Nocturnal polysomnographies (PSG) and SC recordings were performed simultaneously in the sleep laboratory in 51 patients with suspected OSA, and were analysed manually by a blinded investigator. RESULTS: The apnoea/hypopnoea index (AHI) obtained by manual SC analysis correlated closely with that obtained by PSG (r = 0.98), but the correlation with AHI obtained by automatic SC analysis was markedly lower (r = 0.83). When an AHI > or =10 was defined as indicative of OSA, the sensitivity of manual SC analysis was 0.97 and its specificity 1.00. The sensitivity of automatic analysis was 0.83 and its specificity 0.95. CONCLUSIONS: In this experimental setting, the SC showed a very high diagnostic accuracy for the diagnosis of OSA and was able to define its severity precisely. The diagnostic accuracy of manual analysis was found to be superior to that of automatic analysis. Considering the results of this technical evaluation, the SC may also be expected to work reliably when it is used in an ambulatory setting. PMID- 11416254 TI - Characterization of 99mTc-DTPA aerosols for lung permeability studies. AB - BACKGROUND: The interpretation of 99mTc diethylenetriamine pentaacetate (99mTc DTPA) aerosol clearance is based on the hypothesis that the 99mTc-DTPA complex is not altered by the nebulization process. OBJECTIVES: To characterize (1) the radiochemical purity (RCP) of 99mTc-DTPA and the stability of labeling after jet nebulization, and (2) the particle size distribution of the aerosol. METHODS: RCP and stability--the aerosol was driven by oxygen, captured on filters which were eluted and RCP was checked by thin layer chromatography. Particle size distribution--the aerosol was generated using dry air (50 psi) at three different flow rates, i.e. 3 (1 run), 6 (4 runs) and 9 l x min(-1) (4 runs). The mass median aerodynamic diameter (MMAD) and the geometric standard deviation (sigma(g)) were determined using a cascade impactor from the radioactivity counted on each stage. RESULTS: The RCP was more than 95% in all cases. Mean MMAD (+/-SD) was 0.70 microm (+/-0.07) at 9 l x min(-1), 0.93 microm (+/-0.05) at 6 l x min(-1) (p < 0.05) and 1.50 microm at 3 l x min(-1). Mean sigma(g) (+/-SD) was 2.02 (+/-0.08) at 9 l x min(-1), 2.00 (+/-0.16) at 6 l x min(-1) and 1.90 at 3 l x min(-1). CONCLUSION: This study demonstrates (1) that the high RCP of 99mTc DTPA is not affected by jet nebulization, even when using oxygen at a high flow rate, and (2) that when using a flow rate between 6 and 9 l x min(-1), the MMAD remains optimal for peripheral lung deposition. PMID- 11416255 TI - Vocal cord dysfunction: an important differential diagnosis of brittle asthma. PMID- 11416256 TI - Chronic bronchiolitis with associated eosinophilic lung disease (eosinophilic bronchiolitis). AB - We encountered an adult patient with dyspnea, eosinophilia and bronchiolitis. He was diagnosed as having diffuse panbronchiolitis, and was treated with erythromycin for 3 years, but his symptoms had gradually worsened. Bronchoalveolar lavage showed marked eosinophilia, and video-assisted thoracoscopic lung biopsy revealed chronic bronchiolitis with associated eosinophilic lung disease (eosinophilic bronchiolitis). To our knowledge, no case of eosinophilic bronchiolitis has been reported in the literature. PMID- 11416257 TI - Acute dyspnoea resulting from pulmonary oedema as the first sign of a phaeochromocytoma. AB - The day after undergoing neck dissection, a 42-year-old woman developed acute dyspnoea due to pulmonary oedema. Measurements with a Swan-Ganz catheter revealed not only cardiac depression but also a greatly increased peripheral vascular resistance: 5,400 dyn x s x cm(-5)/m2. A phaeochromocytoma with acute cardiac failure leading to pulmonary oedema was considered. Treatment with alpha- and beta-blockers was complicated by severe hypotension and later ventricular fibrillation. Mechanical ventilation was required for 6 days following resuscitation. Investigation of the urine subsequently showed greatly increased catecholamine concentrations, while imaging revealed bilateral adrenal tumours. Our case history shows that acute pulmonary oedema may be the presenting manifestation of a phaeochromocytoma. The pulmonary oedema resulted partly from backward failure following tachycardia, myocyte necrosis and the greatly increased peripheral vascular resistance, and partly from increased permeability of the capillary network in the lungs. PMID- 11416258 TI - Pulmonary alveolar lithiasis in two siblings. AB - Pulmonary alveolar microlithiasis (PAM) is a rare disease of unknown etiology and is characterized by the deposition of calcium phosphate microliths within the alveolar airspaces. We report 2 asymptomatic siblings, a 7-year-old girl and her 13-year-old brother, with PAM. In the girl, chest X-ray and computed tomography revealed diffuse interstitial changes but no uptake of technetium 99m (99mTc) on bone scan was noted in the lung. Microliths stained pink with Papanicolaou dye in bronchoalveolar lavage fluid (BALF) but did not stain with von Kossa. In the brother, characteristic radiological findings and 99mTc uptake in the lung were detected. The microliths stained pink with Papanicolaou in BALF and black with von Kossa as well. We hypothesize that the first case is in the early phase of PAM because of lack of 99mTc uptake. PMID- 11416259 TI - Haemoptysis in a patient with a pulmonary mass and previous tuberculosis. PMID- 11416260 TI - Quantitative on-line analysis of physiological data for lesion placement in pallidotomy. AB - A computerized method of determining the focal point of electrical activity in the pallidum of parkinsonian patients was developed using on-line quantitative physiological data analysis. Thirty patients in a series of 70 were studied in depth. Neuronal activity was recorded from the pallidum using a semi microelectrode. The signal is inspected visually while its average power, characteristic frequency and complexity are computed. The target locus was indicated by the highest level of global activity in the vicinity of the electrode (signal power maximum), maximal signal complexity and minimal characteristic frequency. Most often, the vertical coordinate required correction. The postoperative clinical and imaging results have indicated the effectiveness of this method. PMID- 11416261 TI - The significance of intrathecal opioid therapy for the treatment of neuropathic cancer pain conditions. AB - The effectiveness of intrathecal opioid therapy when applied to different pain mechanisms, in particular neuropathic and nociceptive pain conditions, was studied retrospectively in 43 patients suffering from cancer pain. On the basis of clinical and radiological data, the pain mechanisms were categorized as nociceptive (n = 23) and neuropathic (n = 20). The average duration of treatment of nociceptive pain was 5 months, of neuropathic pain only 2.5 months. The initial median reduction of pain with intrathecal opioid therapy was 77.8% for nociceptive and 61.1% for neuropathic pain. Long-term results with patients suffering nociceptive pain showed a continuing good median pain reduction of 66.7%. Patients suffering from neuropathic pain showed poor long-term results (11.1% median pain reduction). Neuropathic pain in the extremities reacted least to the application of intrathecal opioids. Optimal results were obtained for nociceptive pain in the trunk area of the body. PMID- 11416262 TI - Combined treatment effects after embolization and radiosurgery in high-grade arteriovenous malformations. Case report and review of the literature. AB - In recent years the multimodal therapy concept has been established for high grade intracerebral arteriovenous malformations (AVM). One strategy of a multimodal treatment approach for patients with AVM (Spetzler grade V) is reported. After endovascular embolization, the patient underwent radiosurgical treatment of the remaining nidus with a linear accelerator, after which he developed a left-sided hemiparesis in conjunction with severe edema. Angiographic control 44 months after radiation showed an occlusion of the radiosurgically treated nidus. The remaining nidus could be removed by microsurgery. The combination of embolization and radiosurgery may provoke prolonged edema and permanent neurological deficits due to inflammatory perivascular changes. PMID- 11416265 TI - Does the Concept of a Hypertension Syndrome Apply in the Aging? PMID- 11416263 TI - Brain surgery with image guidance: current recommendations based on a 20-year assessment. AB - Image guidance promotes safe and effective surgical management of a wide array of intracranial diseases. To better define the historical importance of image guidance and to assess the relative contribution of each imaging modality to the safety and efficacy of selected procedures, we reviewed our 20-year experience at a single institution. A retrospective review of our departmental surgical records was performed to identify patients who underwent brain surgery with image guidance between January 1979 and January 1999. We identified the use of intraoperative fluoroscopy, endoscopy, computed tomography (CT), magnetic resonance imaging (MRI), ultrasound, and angiography in 7,388 patients. During this 20-year interval, advances in neuroimaging were translated into the operating room environment. Fluoroscopic guidance received the highest overall rating and was deemed critical for the performance of successful transsphenoidal surgery (n = 436) and effective percutaneous trigeminal neuralgia management (n = 1,121). Ultrasound and angiography both had limited roles; the latter was important to successful outcomes in 64 patients undergoing aneurysm management (n = 64) and arteriovenous malformation Gamma Knife radiosurgery (n = 786). Endoscopy also had a small role but had limited cost. Beginning in 1982, a dedicated operating room CT scanner was used during both morphologic and functional stereotactic surgery (n = 1,749). After 1986, MRI was used increasingly in the management of selected functional and tumor cases (n = 337); despite great versatility for patients undergoing Gamma Knife radiosurgery, the costs were relatively high. Frameless neuronavigation (n = 263) had excellent versatility and was relatively low in cost. During the last 20 years, image guidance techniques have facilitated minimally invasive brain surgery at our institution. The relative merits of all these imaging tools depended mostly on their versatility and relative costs. Major centers currently contemplating the incorporation of image guidance into routine brain surgery need not reproduce our own learning curve. PMID- 11416266 TI - The Systolic Hypertension in the Elderly Program: Implications for the Management of Older Hypertensive Patients. PMID- 11416267 TI - Relation of Age to Left Ventricular Structure, Function, and Systemic Hemodynamics in Normotensive and Hypertensive Employed Adults. AB - To determine the effect of age on left ventricular status and systemic hemodynamics, we evaluated by echocardiogram older (age at or above 60 years, n equals 148) and younger (age is less than 60 years, n equals 472) employed adults. Using World Health Organization criteria, the population was divided into the following blood pressure strata: normotensive (n equals 142), borderline hypertensive (n equals 194), and sustained hypertensive (n equals 284). Older subjects were compared to younger subjects in each blood pressure stratum. Older subjects with borderline hypertension and sustained hypertension had a higher systolic blood pressure and wider pulse pressure than younger individuals. Older subjects with borderline hypertension had higher left ventricular mass index (106.7A+/-28.7 vs 93.9A+/-22.4, P equals 0.03), and greater wall thicknesses and relative wall thicknesses than younger subjects. Older borderline hypertensive patients had greater left ventricular wall thicknesses and mass, but closely resembled both age groups of sustained hypertensive patients in this regard. Older and younger individuals with borderline hypertension had similar mean cardiac outputs, stroke volumes, and peripheral resistances. In contrast, older patients with sustained hypertension had a lower stroke volume, stroke volume index, cardiac output, cardiac index, and fractional shortening, and a higher total peripheral resistance than their younger counterparts. There were no significant relations between age and any echocardiographic measurement in normotensive subjects. Borderline hypertensive individuals exhibited weak relationships between age and left ventricular wall thicknesses (r equals 0.17; 23) and left ventricular internal dimensions (r equals -0.19 and -0.21). Very weak negative relationships were identified between age and cardiac output for both hypertensive groups. In conclusion, increasing age had relatively little effect on cardiac status in either normotensive or sustained hypertensive employed adults, but was associated with greater left ventricular hypertrophy among individuals with borderline hypertension according to the World Health Organization classification. These results are consistent with the independent value of age and left ventricular mass as predictors of hypertensive complications, but also suggest that under some circumstances, greater age or longer exposure to hypertension may magnify the target-organ effects at a given degree of blood pressure elevation. PMID- 11416268 TI - The Need for Outcome-Based Healthcare Research in the Aging: A Tale of Two Procedures. PMID- 11416269 TI - The Rational for the Use of A -Blockers in the Treatment of Congestive Heart Failure. AB - Limited clinical data suggest that in particular types of congestive heart failure, the cautious use of low-dose A -blockers may be beneficial, despite the well-known cardiodepressant effect of these drugs. Downregulation of cardiac A adrenoceptors has been demonstrated repeatedly in various forms of congestive heart failure. This phenomenon is caused by the long-term exposure of these receptors to endogenous noradrenaline (a A -adrenoceptor agonist), which is known to reach markedly elevated plasma concentrations in severe congestive heart failure. As but one example, the downregulation of A -adrenoceptors explains the well-known tachyphylaxis in the inotropic response to A -adrenoceptor agonists such as dobutamine. A beneficial effect of low-dose A -blockers has been shown for metoprolol and bucindolol given for several months. Most studies with such agents have been done in patients with severe congestive heart failure resulting from dilated cardiomyopathy. The beneficial effect of such therapy may be explained by A -adrenoceptor upregulation, thus allowing a better response to endogenous noradrenaline. Alternatively, the therapy may exert its effect by inhibiting tachycardia, which is known to accompany congestive heart failure. PMID- 11416270 TI - The Potential Downside of Treating the Elderly with Hypertension. PMID- 11416271 TI - Pulmonary Embolism: Presentation, Diagnosis, and Treatment in the Aging. PMID- 11416272 TI - Diabetes Mellitus in the Aging: Epidemiologic and Therapeutic Considerations. AB - Diabetes mellitus is prevalent in older persons and a major risk factor for the development of cardiovascular disease. Impaired glucose tolerance without overt diabetes has also been shown to be associated with an increased risk of cardiovascular disease. Elderly patients with non-insulin-dependent diabetes mellitus are at increased risk for both symptomatic and asymptomatic myocardial infarction, and appear to have worse outcomes than nondiabetic persons with these same conditions. Older diabetic persons also have a greater risk than nondiabetics of developing systemic hypertension and stroke. Treatment of diabetes in the elderly can be divided into treatment of the hyperglycemia itself and treatment of other cardiovascular risk factors. It is unknown whether treatment of hyperglycemia in older patients will decrease the risk of cardiovascular and cerebrovascular disease, but it is proposed that treating diabetes along with other commonly associated risk factors for cardiovascular disease may provide the best outlook for such patients. PMID- 11416273 TI - Report from the American College of Cardiology 41st Annual Scientific Sessions, April, 1992, Dallas, TX: Part II. PMID- 11416274 TI - Advance Directives: A Model for Discussing a Neglected Subject with Aged Patients. PMID- 11416275 TI - Screening the Older Patient for an Exercise Program. PMID- 11416277 TI - Inclusion of Elderly Individuals in Clinical Research: An Idea Whose Time has Arrived. PMID- 11416276 TI - Elderly Patients with Late Onset of Coronary Artery Disease: Clinical and Angiographic Findings. AB - A group of elderly patients who did not clinically manifest coronary artery disease until after the age of 65 years were studied. Clinical and angiographic findings of these older patients were compared to findings of patients who had clinical manifestation of coronary artery disease before age 65. In both groups the most common initial manifestation was acute ischemic chest pain (62% vs 75%, NS). However, only 14 (34%) of the 41 elderly patients with chest pain sustained an acute myocardial infarction, compared to 62 (60%) of the 103 younger patients (P is less than 0.05). In 8% of the elderly patients the initial manifestation was acute heart failure not associated with an acute myocardial infarction; none of the younger patients had heart failure without an associated acute infarction (P is less than 0.002). Two percent of the patients over 65 years of age and 4% of the patients under 65 years sustained out-of-hospital cardiac arrest as the initial manifestation of coronary artery disease (NS). Risk factors were more common in the younger patients, with elevated cholesterol levels noted in 24% of the younger patients as compared to only 12% of the elderly patients (P is less than 0.05). Angiographically, elderly patients more commonly demonstrated multivessel coronary artery disease (68% vs 57%, P is less than 0.02) and left main vessel disease (11% vs 2%, P is less than 0.01). Even though multivessel coronary artery disease was common, the majority of elderly patients had normal left ventricular function, as did the younger patients; only 9% of patients in each age group had left ventricular ejection fractions of less than 35%. Treatment varied between the age groups, with coronary artery bypass surgery performed in 38% of the elderly patients as compared to 25% of the younger patients (P is less than 0.05). Only 3% of the patients in each group died during the acute hospitalization, and approximately 95% of the discharged patients in each group were still alive at 24 months mean follow up. We conclude that even though multivessel disease is usually present, coronary artery disease may be clinically silent until the patient is quite old. Left ventricular function is usually well preserved in these elderly patients, and with therapy their prognosis is excellent. PMID- 11416278 TI - Treatment of Very Elderly Hypertensive Patients in Long-term Care Facilities. PMID- 11416280 TI - Pathogenesis of Sodium and Water Retention in Congestive Heart Failure: Therapeutic Implications. PMID- 11416279 TI - Do Older Patients Need a Different Approach to Rehabilitation After Coronary Revascularization? AB - Despite the increasing size of the population over the age of 65 years and increasing use of coronary revascularization procedures in the elderly population in the developed countries of the world, very limited information is available about the rehabilitative needs of people in this age group. We conducted a study to compare the baseline characteristics, before scheduled coronary revascularization, of patients over and under 65 years of age, in order to determine whether a need exists for a different approach in the rehabilitation of elderly patients. One hundred eighty-two consecutive patients with angina pectoris, who were scheduled to undergo isolated coronary artery bypass grafting or percutaneous transluminal coronary angioplasty for the first time, were examined at the time of coronary angiography with respect to sociodemographic data, extent of coronary artery disease, risk-factor levels, and psychosocial status with the help of self-administered questionnaires, including the Nottingham Health Profile. Eight-four patients (52 male and 32 female) aged 65 years and over were compared to 98 (84 male and 14 female) patients younger than 65 years. Three-vessel disease was significantly more frequent (73% vs 44%, P equals 0.001) in the older age group than in the younger. Risk-factor levels were similar in both age groups. With regard to quality of life, the elderly patients, as expected, experienced less impairment of function in their work (since 93% were retired) and more limitation in their mobility. Otherwise, there were no significant differences between the two age groups in the parameters examined. We conclude from this baseline study that elderly patients do not need a different approach to rehabilitation, provided there is a flexible, patient-specific approach to multifactorial intervention after coronary revascularization. PMID- 11416281 TI - From Left Ventricular Dysfunction to Heart Failure in the Elderly Patient. AB - The pathophysiology of the syndrome of congestive heart failure (CHF) is different in elderly patients compared to middle-aged subjects. When the syndrome of CHF predominantly results from left ventricular (LV) systolic dysfunction, peripheral mechanisms in elderly patients are less apt to compensate for the decline in LV performance due to deconditioning of the skeletal muscles, decreased vasodilatory response to exercise, and reduced capacity to excrete sodium. These all develop with age. LV diastolic dysfunction, i.e., LV filling impairment, appears to be a more frequent cause of CHF in elderly patients than in middle-aged subjects. Hypertrophy of the cardiac myocyte is associated with a prolongation of the calcium transient which, in turn, results from a decreased concentration of the enzyme responsible for reuptake of calcium in the sarcoplasmic reticulum. Cardiac myocyte hypertrophy results from a steady loss of myocytes and a progressive rise in arterial impedance which are both observed with aging. Notwithstanding subendocardial myocardial ischemia which is frequently associated with LV wall hypertrophy, prolongation of the calcium transient leads to impaired LV wall relaxation and the syndrome of LV diastolic dysfunction. The goals of therapy are also different in elderly patients compared to middle-aged patients. In elderly patients, one is less concerned about restoring a near normal exercise capacity, and aims at preventing acute episodes of decompensation which frequently accompany excessive sodium intake. Therefore, strict adherence to diet and careful titration of the dosages of medication to renal function are important considerations when treating CHF in elderly patients. In particular, cardiac glycosides, which should be avoided with CHF due to LV diastolic dysfunction, should be carefully titrated to creatinine clearance and body weight when treating elderly patients with LV systolic dysfunction in sinus rhythm. Similarly, the dosages of most angiotensin-converting enzyme inhibitors should be reduced in elderly patients. PMID- 11416282 TI - Studies of Left Ventricular Dysfunction (SOLVD). AB - Given the increasing incidence and dismal prognosis in congestive heart failure, it is not surprising that strenuous efforts have been made to limit the morbidity and mortality that result from this syndrome. The National Institutes of Health designed and implemented the Studies of Left Ventricular Dysfunction (SOLVD) to assess the effect of therapy with enalapril, an angiotensin-converting enzyme inhibitor, on all cause mortality in a population with left ventricular dysfunction (i.e., ejection fraction at or below 35%). Important secondary objectives included the effects of this therapy on cause-specific mortality, development of heart failure, and hospitalization for congestive heart failure. Patients within each of two arms of the trial were randomized on the basis of the administration (treatment arm) or lack (prevention arm) of concomitant therapy for congestive heart failure. The trial was double blind and placebo controlled. The mean follow-up period for the combined arms was 39.2 months, during which time there was a 16% risk reduction in all-cause mortality in the treatment arm of the trial (P equals 0.008) and an 8% risk reduction in the prevention arm (P equals NS). In the prevention arm, however, there was a 12% reduction risk for cardiovascular mortality (P equals 0.12) and a 37% reduction in risk of progression to heart failure (P is less than 0.001). Thus, the results of SOLVD indicate that therapy with an angiotensin-converting enzyme inhibitor has beneficial effects on morbidity and mortality that are apparent in a broad spectrum of patients with left ventricular dysfunction. PMID- 11416283 TI - Aspects of the Use of Angiotensin-Converting Enzyme Inhibitors and Calcium Antagonists in Treatment of Heart Failure in the Older Patient. AB - The syndrome of congestive heart failure may result from either systolic or diastolic dysfunction of the left ventricle. Diastolic left ventricle dysfunction is particularly common in the geriatric age group, and is associated with left ventricular hypertrophy resulting from aging and hypertension. The clinical differentiation of these two patterns is important in understanding the pathophysiologic process and in selecting appropriate therapy. Angiotensin converting enzyme (ACE) inhibitors are useful in systolic dysfunction, both in improving clinical manifestations of reduced cardiac output and in actually prolonging survival. ACE inhibitors are also beneficial in diastolic heart failure by promoting regression of left ventricular hypertrophy, thus improving diastolic physiological function. Calcium antagonists improve diastolic function by reducing blood pressure of hypertensive subjects, reducing left ventricular mass, and theoretically, by facilitating the energy-dependent transport of calcium ions from the actin-myosin complex into the sarcoplasmic reticulum. However, because of the negative inotropic properties of the calcium antagonists, they should be used cautiously, if used at all, in patients with significant systolic dysfunction, at least until the results of clinical trials using these drugs in systolic congestive heart failure are available. PMID- 11416284 TI - Should Age Influence Our Choice of Antihypertensive Medications. PMID- 11416285 TI - Age Related Complications of Pulmonary Angiography for Acute Pulmonary Embolism. AB - The relation of age to complications of pulmonary angiography was assessed among 1111 patients who underwent pulmonary angiography for acute pulmonary embolism in the collaborative study of the Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED). The quality of pulmonary angiograms was similar among all age groups in the study. Neither major nor nonmajor complications occurred with a greater total frequency among elderly patients than among young patients. Manor and nonmajor renal failure, however, were more frequent among patients 70 years of age or older, and this was the most frequent complication of angiography in this age group. Among patients 70 years of age and older, renal failure occurred in 7 of 281 (2.5%), as opposed to 6 of 830 (0.7%) patients 69 years old or younger (P is less than 0.05). Patients with either major or nonmajor renal complications were older than patients with no renal dysfunction following pulmonary angiography (74A+/-13 yr vs 57A+/-17 yr) (P is less than 0.001). Most of the patients who died or sustained major nonfatal cardiopulmonary complications, irrespective of age, were in critical condition with severely compromised cardiopulmonary function prior to angiography; the severity of the underlying condition rendered these patients vulnerable to the procedure. In conclusion, high-quality pulmonary angiography can be done safely on elderly patients. The risks of pulmonary angiography are sufficiently low to justify it as a diagnostic tool in the appropriate clinical setting, even in elderly patients. Clinical judgment is important in selecting patients and assessing their risk. PMID- 11416286 TI - Age and the Hemodynamic Response to Posture and Exercise. AB - The effects of age on hemodynamic variables at rest, in response to orthostatic challenges, and during exercise were assessed from overall weighted statistical summaries of reports in the medical literature. In normotensive subjects at supine rest, cardiac output and stroke volume (p is less than 0.001), but not heart rate, declined with aging, whereas systemic vascular resistance increased (p is less than 0.001). The results were similar in hypertensives, but the slope of the change in cardiac output was steeper than in normotensives (p is less than 0.01). This age-related hemodynamic pattern was also observed in longitudinal follow-up studies. The orthostatic fall in stroke volume and rise in heart rate became less pronounced with increasing age (p is less than 0.05), but the postural increase in systemic vascular resistance was, on average, not related to age. Though the hemodynamic studies in healthy individuals observed that blood pressure is relatively well maintained when older subjects assume the upright position, population studies suggest that an increasing number of these subjects show a postural fall in blood pressure. This is overwhelming evidence that the oxygen uptake at peak exercise decreases with age, mainly related to a lesser peak cardiac output and heart rate (p is less than 0.001). PMID- 11416287 TI - Report from the American College of Cardiology: 42nd Annual Scientific Sessions, March 14-18, 1993, Anaheim, CA: Part I. PMID- 11416289 TI - Risk Factors in the Elderly: A View From Framingham. PMID- 11416288 TI - Hypercholesterolemia and Coronary Risk in the Elderly: The Framingham Study. AB - In a follow up of elderly Framingham men and women, and after multivariate adjustment, the total/high-density lipoprotein-cholesterol ratio remained highly associated with the incidence of coronary heart disease in both sexes, whereas total cholesterol level was associated with coronary heart disease only in women. Whereas total cholesterol values decline in the very elderly, and the association of coronary heart disease with total cholesterol level alone is weaker in the elderly than among those of middle age, the total/high-density lipoprotein cholesterol ratio remains a strong predictor of coronary heart disease. In assessing the elderly for coronary risk, lipid measurements should include high density lipoprotein-cholesterol and cholesterol determinations. PMID- 11416290 TI - Cardiovascular Risk Factors in the Elderly in Italy. PMID- 11416291 TI - Isolated Systolic Hypertension, Morbidity, and Mortality: The SHEP Experience. PMID- 11416292 TI - Need for Physical Activity in the Elderly Population. PMID- 11416293 TI - Hormone Replacement. PMID- 11416294 TI - Oxidative Modification of LDL in the Pathogenesis of Atherosclerosis. PMID- 11416295 TI - Secondary Prevention of Cardiovascular Disease. PMID- 11416296 TI - Sodium Restriction in Elderly Patients: Does It Have a Role? PMID- 11416297 TI - Permanent Cardiac Pacing in the Elderly Patient: Is Selection of the Pacing Mode Important? AB - Most pacemaker recipients are elderly, and advanced age does not constitute a contraindication to the implantation of a permanent pacemaker. However, pacing in the older patient should no longer be regarded simply as a way to prevent Stokes Adams attacks or life-threatening bradyarrhythmia. A VVI pacemaker for everyone is now inappropriate, especially in active and otherwise healthy older patients. Selection of the optimal devise or pacing mode in older patients requires an understanding of how aging affects the cardiovascular system (left ventricular diastolic dysfunction) and the natural history of disease, especially sick sinus syndrome after pacemaker implantation. Many retrospective and nonrandomized studies focusing mostly on the sick sinus syndrome have indicated that atrial based pacing improves the quality and duration of life when compared to single lead ventricle pacing. The lower incidence of chronic atrial fibrillation in patients with atrial-based pacemakers compared to VVI devices is of great importance in the elderly, in whom this arrhythmia is associated with greater morbidity and mortality than in younger individuals. The decreased mortality associated with atrial-based pacing compared to single lead ventricular pacing seems to favor patients older than 70 years. There is now a growing realization that implantation of dual chamber pacemakers in the elderly is cost-effective on a long-term basis by avoiding or reducing the complications associated with single lead ventricular pacing, thereby reducing the need for repeated hospitalizations and medical care. PMID- 11416298 TI - The Cerebral Circulation in the Elderly: The Influence of Age, Vascular Disease, and Antihypertensive Treatment. AB - Cerebral blood flow (CBF) and cerebral metabolic activity tend to decrease in old age. The effect of age per se on the regulation of CBF is not readily separated from the effect of age-associated diseases such as atherosclerosis, multi-infarct dementia, and diabetes mellitus, which may lead to an impairment of CBF autoregulation. Stroke may be prevented effectively by antihypertensive treatment in otherwise healthy elderly patients, but care should be taken to prevent overtreatment with consequent cerebral ischemia. The risk of this may be especially great in elderly patients with hypertension who have postural hypotension. PMID- 11416300 TI - Report from the American Geriatrics Society and American Federation for Aging Research Annual Meeting, November 15-19, 1993, New Orleans, LA. PMID- 11416299 TI - What's Different About Syncope in the Aged? AB - Syncope is a common and highly morbid syndrome with unique features in the elderly population. Syncope in the elderly is more likely due to the accumulation of multiple pathological conditions which threaten cerebral oxygen delivery, then to single diseases. Furthermore, age-related changes in blood pressure homeostasis make older patients more susceptible to hypotension during situational stresses such as posture change, eating a meal, or taking medications. An age-related impairment in baroreflex sensitivity may increase the risk of hypotension, but paradoxically protect older patients from the development of vasovagal syncope. Finally, the elderly are at greater risk of cardiovascular conditions such as myocardial infarction, valvular heart disease, and arrhythmias which often produce syncope. The evaluation of syncope in an elderly patient requires careful assessment of blood pressure during activities associated with a syncopal event, as well as attention to each of the contributing pathologic abnormalities that increase the risk of this syndrome. PMID- 11416301 TI - When is a Cough Just a Cough? The Issue of Quality of Life and Its Assessment. PMID- 11416302 TI - Noninvasive Coronary Risk Stratification of Elderly Patients. AB - The prognostic value of stress thallium-201 myocardial perfusion imaging has not been defined in an elderly (at or above 70 years) population. To this end, we studied 468 consecutive elderly patients undergoing either Bruce protocol exercise stress (n equals 120) or intravenous dipyridamole stress (n equals 348) with quantitative planar thallium-201 imaging. These patients were followed for at least 2 years after testing to determine their rates of cardiac events (cardiac death, myocardial infarction, coronary revascularization). There were no stress-related complications in either subgroup. A 10% cardiac event rate (6 deaths and 6 myocardial infarctions) was observed in the exercise subgroup. Survival without cardiac events was associated with greater exercise duration (5.6A+/-2.4 vs 3.1A+/-2.4 min; P is less than 0.001) and peak exercise heart rate (131A+/-18 vs 120A+/-19 bpm; P is less than 0.05). Multivariate analysis identified the combination of peak exercise at or below Stage 1 and thallium-201 perfusion defects as significant predictors of cardiac events (relative risk equals 5.3 at 1 year). Sixty-four percent of elderly patients were successfully stratified into very low and high risk subgroups, with annual cardiac event rates of less than 1% and greater than 15%, respectively. The cardiac event rate in dipyridamole stress patients was 22% (24 myocardial infarction, 52 death, 42 revascularization). The cardiac event rate was significantly lower (5%) in 150 patients with a normal dipyridamole thallium-201 study (P is less than 0.001). Clinical univariate predictors of cardiac events were previous myocardial infarction, congestive heart failure symptoms, hypercholesterolemia and diabetes (all P equals 0.05). A fixed, reversible, or combined thallium defect pattern was correlated with cardiac death or myocardial infarction (P is less than 0.05). Multivariate analysis demonstrated that the presence of an abnormal dipyridamole thallium study was the single-best predictor of cardiac events (relative risk equals 7.2; P is less than 0.01) We conclude that exercise and dipyridamole thallium-201 myocardial imaging are powerful independent noninvasive techniques for prognostication in the elderly patient population, a group with the potential for advanced coronary artery disease and a high risk of cardiac events. PMID- 11416303 TI - Coarctation of the Aorta in the Elderly: Case Report and Review of the Literature. AB - Coarctation of the thoracic aorta is a rare condition in the elderly population. Approximately 90% of untreated patients die by the age of 50. The complications of persistent coarctation include cerebrovascular accidents, congestive heart failure, intracranial aneurysms, endocarditis, aortic valve disease, myocardial infarction, aortic rupture, and aortic dissection. We report the case of a 62 year-old man who presented with complete aortic occlusion distal to the left subclavian artery. The effect of antihypertensive therapy on renal function in patients with coarctation is discussed, and the literature regarding elderly patients with coarctation of the aorta is reviewed. PMID- 11416304 TI - Cardiac Surgery in Patients At or Above 75 Years Old: Analysis of Perioperative and Long-term Outcome. AB - BACKGROUND: The peri-operative and long-term outcome of 211 consecutive patients at or above 75 years of age undergoing cardiac surgery between 1987-1990, was assessed. 79.6% of patients underwent CABG, 8.5% underwent both CABG and valvular surgery, 8.1% underwent only valvular surgery and 3.8% underwent other procedures. RESULTS: Survival curve analysis revealed no intra-operative deaths and a 30 day, 1,3, and 5 year overall survival rate of 93.3%, 89.2%, 77.8% and 64.6% respectively. 30.8% of patients had major peri-operative complications. Transient post-operative atrial fibrillation occurred in 47% of patients. There was a non-statistical trend towards increased risk of post-operative stroke in patients with transient post-operative atrial fibrillation. Early cardiothoracic re-operation was required in 8.5%. 91.9% of patients were followed up at a mean of 36.5A+/-18.2 months post-operatively. 94.1% lived at home, 96.3% were ambulant, 96.3% were NYHA class I or II. Previously undetected malignancy was he cause of mortality in 10/45 (22%) patients who died after the thirtieth post operative day, during the follow-up period. CONCLUSIONS: Chronologic age alone is not a contra-indication to cardiac surgery. Standard selection criteria with careful consideration of risk-benefit ratio, quality of life, presence of comorbid disease and health care costs should be factored in the decision process regarding cardiac surgery in the elderly. Particular attention should be paid to screening for malignancy and to management of transient post-operative atrial fibrillation. PMID- 11416305 TI - Nutrition and the Aging Process: A Population Study. AB - Highly significant correlations exist in the age range 35 to 84 years between log mortality from all causes and age, obtained from 33 countries (mean r equals 0.999 in men and 0.997 in women). Populations with a high life expectancy are characterized by a low intercept and a high slope of the equation between log mortality and age. The slope and intercept of the equation obtained from different countries are highly intercorrelated, pointing to a maximum life expectancy at the population level of about 104 years in men and 108 years in women. The slope of the equation between log mortality and age can be considered as an expression of the resistance against aging of a given population. Significant positive correlations exist between the level of saturated fat intake and the determinants of mortality at the population level. These correlations are more evident in men compared with women. The results offer evidence for a detrimental influence of saturated fat on the aging process in populations. PMID- 11416306 TI - An Unusual Cause of Dyspnea, Hypoxemia and Pedal Edema in an 85-Year-Old Woman. AB - Atrial septal defect (ASD) is one of the most common causes of congenital heart disease manifested in adulthood. Among the three kinds of ASD, the clinical features of secundum-type ASD have been well-characterized in patients less than 40 years of age, and account for two-thirds of all such defects (sinus venosis and ostium primum are the second- and third-type defects, respectively). More than 90% of patients with a secundum ASD die before the age of 60. However, a subgroup of patients may or may not develop symptoms until late adulthood, presenting with atrial fibrillation, dyspnea and heart failure as the first manifestation of secundum ASD. In the last 30 years, six octogenarians presenting similarly with ASD have been described in the literature. The diagnosis requires a high index of clinical suspicion combined with careful inspection of the electrocardiogram and chest radiograph for signs of right ventricular volume overload. Transthoracic echocardiography and color Doppler interrogation of the atrial septum combined with bubble contrast should confirm the presence of secundum ASD in the majority of elderly patients presenting with this unusual clinical entity. PMID- 11416307 TI - Atrial Fibrillation in the Elderly: A Review. AB - Atrial fibrillation is the most common cardiac arrhythmia in the elderly. It is associated with significant morbidity and mortality due to hemodynamic and cardioembolic complications. The incidence of stroke in elderly patients is 5 times higher than in patients in sinus rhythm. Atrial fibrillation should be actively treated in the elderly. Treatment should be directed toward the correction of reversible factors, control of ventricular response rate, restoration and maintenance of sinus rhythm, and prevention of cardioembolic events. Treatment of atrial fibrillation in the elderly should be individualized with careful regard for risk-benefit ratio. PMID- 11416308 TI - Can Blood Pressure in the Elderly be Reduced? Findings From a Long-term Population Survey in Japan. AB - To examine whether blood pressure (BP), a major risk factor of cardiovascular disease, can be controlled in the elderly as well as in middle-aged persons, we analyzed the data of observational studies on trends for BP and cardiovascular disease incidence in a northeast rural community of Japan. This community was the subject of an ongoing hypertension control program that was initiated in 1963. A significant decline in BP levels was noted in each sex-age group between 1963 and 1966 and 1987 and 1991. The decline was greater in older persons compared with younger individuals. The BP decline was attributable to an increase in antihypertensive medication use, beginning in the 1970s. We compared BP levels of untreated offspring and parents when both were 40 to 49 years old. Blood pressure levels were significantly lower in the offspring than in parents. This result and the large downward shift of BP distribution in the second decade of follow-up suggested that the improvements in diets and other environmental factors contributed to the BP decline. Between 1964 and 1968 and 1989 and 1992, stroke incidence declined 70% to 79% for all sex and age groups (40-69 and greater than 70 years). The number of totally dependent stroke patients decreased in both the middle-aged and the elderly between 1976 and 1991. The decline in stroke mortality tended to be larger in the surveyed community than in adjacent communities. These results indicated that hypertension control is effective in preventing stroke in the elderly as well as in the middle-aged. PMID- 11416309 TI - Health and Social Consequences of Prolonging Life, With Special Reference To Cardiovascular Diseases: The Italian Situation. AB - During the last century, the reduction of birth and all-cause mortality rates have resulted in an increase in the life expectancy and the proportion of the elderly population in many countries. For example, elderly women represent more than 16% and elderly men comprise more than 12% of the population in Italy. The health consequences of these demographic shifts in population is important, because older people experience reductions in some physiologic functions and in increased incidence of chronic, mainly cardiovascular, diseases. Data derived from a survey of men between the ages of 71 and 91 years, who are the survivors of the original rural samples of the Italian section of the Seven Countries Study, show the large contribution of cardiovascular diseases to the long list of chronic conditions. The indicators of self-sufficiency, dementia, and of the self perception of poor health proved much more common among subjects with cardiovascular diseases than in those with other chronic conditions. PMID- 11416310 TI - Rational for Treatment of Hypertension in the Elderly. AB - Hypertension is a powerful predisposing factor to the major cardiovascular problems common to the elderly, doubling the risk. This increased risk of cardiovascular events exceeds that associated with the existence of hypercholesterolemia, diabetes, and smoking. The decrease in risk ratio with advancing age is offset by the increased cardiovascular disease incidence, resulting in a greater attributable risk, which makes treatment of hypertension more cost effective in the elderly. The prevalence of hypertension is high in the elderly, with about 50% afflicted; 65% of those patients have isolated systolic hypertension (ISH). Hypertension in the elderly often occurs concomitantly with hypercholesterolemia, diabetes, obesity, hypertriglyceridemia, and left ventricular hypertrophy, all of which enhance the risk. Also, 25% of these patients already have manifest cardiovascular disease. These associated cardiovascular conditions and risk factors determine the urgency for treatment and choice of optimal therapy. Antihypertensive trials indicate greater benefit of antihypertensive therapy in the elderly than in the young and have been shown to clearly benefit those with ISH. Thus, hypertension is dangerous in the elderly, is readily controlled, and treatment can provide great benefit in preventing cardiovascular sequelae. PMID- 11416311 TI - Cardiac Electrophysiology in the Older Population. AB - There is increasing evidence that significant primary alterations of cardiac electrophysiologic properties occur with the normal aging process. The age related changes in cardiac electrophysiologic parameters are seen as alterations in the surface electrocardiogram, depressed sinus and atrioventricular nodal function, and increased refractoriness of the cardiac tissue. The mechanisms underlying these changes may consist of structural changes of the conduction system, autonomic nervous system dysfunction, or changes in cellular electrophysiology. As a result of these changes, the responsiveness of the aging heart to cardiotropic drugs is different from that of the younger heart. Elderly subjects are prone to certain electrophysiologic syndromes that merit special consideration. PMID- 11416312 TI - Exercise and Survival in the Very Old. AB - Risk factors that are strongly associated with cardiovascular events in middle age are less predictive in the elderly. These reduced associations are most evident after about age 75 years. Exercise has been associated with survival in middle-aged men; data are sparse on its relation to survival in the very old. This association was tested in a community-dwelling cohort of 440 men and 457 women aged 75 years or older who were followed prospectively for an average of 5 years. In age-adjusted proportional hazards models, current regular exercise at least 3 times per week was strongly associated with survival in men (relative risk for death 0.54, P is less than.01) and women (relative risk 0.45, P is less than.01). Results were unchanged after adjustment for physical and emotional health status, or when persons with heart disease at baseline were excluded. Results were also unchanged after adjustment for change in exercise level compared with 10 years ago, and for conventional cardiovascular risk factors (blood pressure, lipid concentration, glucose levels, and smoking). PMID- 11416313 TI - Smoking, Lung Function, and Atherosclerosis in the 5,000 Elderly Participants of the Cardiovascular Health Study. AB - The Cardiovascular Health Study (CHS) is an observational study of heart disease and stroke designed to evaluate risk factors and noninvasive measures and to describe and predict atherosclerotic events in older adults. Five thousand two hundred one individuals ages 65 or older were recruited from a stratified random sample of Medicare recipients from 4 US communities. This review of cross sectional data from the CHS baseline examination describes the cigarette smoking habits of elderly persons and the relationships of smoking to lung function (spirometry) and atherosclerosis, as noninvasively measured by the ankle-arm index (AAI) and carotid ultrasonography. Only 10% of the men and 13% of the women were current smokers, and about half were former smokers. Forced expiratory flow (FEV1) was about 20% lower in current smokers when compared with never smokers. Current and former smoking were strongly associated with an increased risk for an abnormal AAI. Common and internal carotid artery walls were thicker and stenosis more common in current smokers and former smokers, when compared with never smokers. Analysis of long-term follow-up morbidity and mortality data from the CHS cohort should provide even stronger evidence of the effects of smoking in the elderly. Vigorous efforts should be made to persuade elderly smokers to quit. PMID- 11416314 TI - Hypertension in the Elderly. AB - In this review article, the authors cover the pathophysiology of elevated blood pressure in the elderly. Isolated systolic hypertension and pseudo-hypertension are noted. Some of the large treatment trials are discussed. Nonpharmacologic and pharmacologic management are covered. Neurologic aspects of hypertension in this population are discussed in some detail. Additional considerations regarding medication choices in the last part are noted. PMID- 11416315 TI - Peripheral Vascular Disease in the Elderly. AB - This paper is reproduced with permission from the book, Cardiovascular Disease in the Elderly, edited by Donald D. Tresch, MD and Wilbert S. Aronow, MD and published by Marcel Dekker, Inc. PMID- 11416316 TI - Electrophysiologic Studies in Octogenarians. AB - To assess the risk-benefit of electrophysiologic studies (EPS) in very elderly patients, we studied 104 consecutive patients 80 years or older who underwent EPS during hospitalization. Seventy-two percent of the patients had a history of coronary artery disease, and 7% had an acute myocardial infarction during the hospitalization. Other cardiac disorders included valvular heart disease (17%) and cardiomyopathy (12%). Abnormal systolic left ventricular function was common, with a left ventricular ejection fraction of 30% or less in 25% of the patients. The most common indication for EPS was unexplained syncope or presyncope (62 patients). Other indications for EPS included evaluation of cardiac arrest (9 patients), sustained monomorphic ventricular tachycardia (7), nonsustained ventricular tachycardia (22), wide QRS tachycardia (1), symptomatic sinus bradycardia (2) and palpitations (1). Thirty-eight patients (37%) demonstrated EPS findings considered to be responsible for the patients' symptoms. Specific clinical characteristics of the patients were not predictive of abnormal EPS findings. After EPS, 60% of the patients had a change in therapy; more than 75% of the patients who presented with sustained ventricular tachycardia or who had survived cardiac arrest and 58% of patients who presented with syncope had a change in therapy after EPS. Complications secondary to EPS occurred in only 3 patients; mortality or neurological complications were not associated with the procedure. We conclude that EPS can be performed safely in very elderly patients and that the procedure is beneficial in that therapy is modified on the basis of the EPS findings. PMID- 11416317 TI - The Elderly Diabetic Patient With Hypertension: Guidelines for Prescribing Antihypertensive Therapy. AB - Diabetes mellitus and hypertension commonly coexist in elderly patients. The benefits of antihypertensive therapy have been clearly defined in the elderly population, but the choice of antihypertensive agents must be based on careful patient evaluation. The elderly patient with diabetes requires evaluation of metabolic and cardiovascular status before therapy is instituted. Antihypertensive therapy should be initiated only after the potential for life style modifications has been exhausted. The goals of drug therapy are to reduce blood pressure without worsening underlying disease processes or increasing cardiovascular risk factors. Other important considerations in the elderly population include the age-related decline in renal and hepatic function, which may reduce metabolic capacity, the potential for polypharmacy, medication cost, and compliance issues. PMID- 11416319 TI - Aging Effects on the Vasculature in Health: Risk Factors for Cardiovascular Disease. PMID- 11416320 TI - Isolated Systolic Hypertension in the Elderly. PMID- 11416321 TI - The Effect of Normative Aging on the Cardiovascular System. PMID- 11416323 TI - Coronary Heart Disease in the Elderly: Management Considerations. PMID- 11416322 TI - Left Ventricular Diastolic Function in the Elderly. PMID- 11416325 TI - Working Group Report On Hypertension in the Elderly. PMID- 11416324 TI - Echocardiographic and Therapeutic Approach to Heart Failure in the Elderly. AB - In most elderly patients, cardiac failure is associated with multiple cardiac pathologies, and the most common underlying abnormality is ventricular systolic dysfunction with reduced ejection fraction. In patients without cardiac enlargement, diastolic dysfunction may be predominant and left ventricular ejection fraction may be normal. Echocardiography is particularly suitable to evaluate changes in cardiovascular function that accompany age either as changes in disease patterns or as changes resulting simply from aging. The effects of digitalis on systolic and diastolic dysfunction are briefly reviewed, as well as the effects of diuretics, angiotensin-converting-enzyme inhibitors, and anticoagulant drugs. PMID- 11416326 TI - Coronary Angioplasty of an Anomalous Coronary Artery in the Elderly: Case Report and Review of the Literature. AB - Newer developments in biotechnology and increasing operator experience have broadened the clinical applicability of percutaneous transluminal coronary angioplasty. These advances have allowed the treatment of patients with variant and anomalous coronary arteries and the treatment of an increasingly elderly population. A case of successful percutaneous transluminal coronary angioplasty in a 90-year-old man with unstable angina and an anomalous origin of the left main coronary artery is reported. PMID- 11416327 TI - Cardiovascular Risk Factors and Age: A Cross-Sectional Survey of Danish Men and Women From the Glostrup Population Studies, 1991. AB - We report the age- and gender-related prevalence of risk factors for coronary heart disease (CHD) in the third screening of the Danish MONICA population, 1991. The study population comprised all inhabitants of 11 municipalities in the outskirts of Copenhagen. Using the Central Person Register (CPR), random samples of exact age groups born in 1921, 1931, 1941, 1951, and 1961 were invited. A total of 1,008 men and 1,015 women were screened (participation rate, 70%). The Framingham coefficients were used as weighting factors to compare combined risk in each age group. Systolic and diastolic blood pressure, total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), and triglyceride levels in serum all increased with age of the cohorts in women. In men, TC concentrations decreased but serum HDL levels increased from cohort age 50, thus bringing TC as well as the TC/HDL ratio statistically below female levels at age 60 and 70. Reasons for this observation are discussed, and include high physical activity, alcohol consumption and selective mortality in the elderly men. PMID- 11416328 TI - Images in Geriatric Cardiology - A Complication of Acute Myocardial Infarction. PMID- 11416330 TI - The Importance of Cardiovascular Risk Factors in the Elderly. AB - The cardiovascular risk factors in the elderly are much the same as those that predispose middle-aged candidates for cardiovascular disease. Some risk factors, such as blood lipids, impaired glucose tolerance, fibrinogen, and uric acid, are associated with lower risk ratios in advanced age, but this lower relative risk is offset by a high absolute risk. Thus, cardiovascular risk factors remain relevant at elderly age. Since the incidence of cardiovascular disease and the prevalence of the predisposing risk factors is so great in the elderly, the attributable risk is large, and the short-term potential benefit of treatment is actually greater in the elderly than in the middle aged. Preventive measures, as well, are useful in reducing the risk of disease in the elderly. Therefore, efforts at primary and secondary prevention should be considered in the elderly they should not be neglected simply because of their age. PMID- 11416331 TI - Pharmacologic Treatment of Hypertension in the Elderly. PMID- 11416332 TI - Epidemiology and Prevention of Cardiovascular Disease in the Elderly Population in Germany. PMID- 11416333 TI - American Heart Association 67th Annual Scientific Sessions Abstract Reviews, Dallas, Texas, November, 1994. PMID- 11416334 TI - Heart Disease in the Elderly. PMID- 11416336 TI - Cardiac Surgery in Elderly Patients. PMID- 11416335 TI - Coronary Heart Disease at Elderly Age: Determinants of Clinical Decision-Making. PMID- 11416337 TI - Angioplasty in the Elderly Patient. PMID- 11416338 TI - Approach to the Elderly Patient With Cardiac Disease: Role of the Geriatric Specialist Within a Multidisciplinary Team. PMID- 11416339 TI - Images in Geriatric Cardiology - Aortic Stenosis. PMID- 11416340 TI - Cardiac Pacemakers and Implanted Defibrillators in Elderly Patients. PMID- 11416341 TI - Aging and Atherosclerosis: Changing Considerations in Cardiovascular Disease Prevention as the Barrier to Immortality is Approached in Old Age. AB - Of all the risk factors to cardiovascular disease (CVD), age is the most powerful: CVD incidence and prevalence rise progressively at all ages beyond young adulthood. This reflects the central role of time, and hence duration, in the atherogenic process. It also reflects age-related changes in physiology - notably alterations in body mass and composition favoring increased adiposity and in sex hormone secretion (combining adverse effects of androgens on lipoprotein lipid levels in males, lowering HDL, and of the decline in estrogens in postmenopausal females, raising LDL). The interactions among the passage of time, these physiological changes and perhaps aging per se, and pathological forces such as cigarette smoking, hypertension, and genetically determined dyslipoproteinemia conspire to accelerate the rate of atherogenesis. Thus clinical atherosclerosis and its complications rise exponentially with increasing age in the population at large. However, the relationship between dyslipoproteinemia and CVD risk in the individual patient actually declines with advancing age. This apparent paradox reflects confounding introduced by the advent of disease processes that cause wasting and inflammation such as cancer, infection, diabetes, trauma, and even CVD that actually lower lipid levels, frequently to the level of hypocholesterolemia. Thus, while with age the population-attributable risk of hypercholesterolemia (and/or low HDL) rises, the cholesterol-attributable risk in the individual falls. As a result the prescription of lipid-lowering therapy in elderly patients requires exquisite individualization: patients most likely to benefit are those with existing CVD (i.e., in secondary prevention) who nevertheless enjoy robust health and are highly motivated to comply with demanding regimens of diet and exercise plus drugs where needed to reach target LDL levels (less than 100 mg/dl). At the other extreme are those least likely to benefit: patients who are frail and failing from CVD or other wasting diseases of old age that present a more immediate threat to survival. PMID- 11416342 TI - 1995 ACC Meeting Abstract Reviews Prepared Exclusively for The American Journal of Geriatric Cardiology. PMID- 11416343 TI - Benefits of Cardiac Rehabilitation and Exercise Training in Older Persons. AB - Despite the well-proved benefits of cardiac rehabilitation and exercise training, older persons are frequently not referred to or vigorously encouraged to pursue this therapy after major coronary heart disease (CHD) events. Therefore, we determined the effects of this therapy on plasma lipids, indices of obesity, and exercise capacity in older CHD patients compared with the benefits obtained in a younger cohort. At baseline, the older persons had lower body mass indices (BMI), triglycerides levels, and estimated metabolic equivalent (METs), and the elderly had higher levels of high-density lipoprotein cholesterol (HDL-C). Most other parameters were statistically similar in the older and younger patients. After cardiac rehabilitation, the elderly had significant improvements in METs, BMI, percent body fat, HDL-C, and low-density lipoprotein cholesterol (LDL-C)/HDL-C but not in total cholesterol or LDL-C. However, improvements in all of these parameters were statistically similar in older and younger patients. We conclude that despite baseline differences, improvements in exercise capacity, obesity indices, and lipid levels were statistically similar in older and younger patients enrolled in formal, phase II, cardiac rehabilitation and exercise training programs. Therefore, our data emphasized that the elderly should not be categorically denied the benefits of vigorous secondary CHD prevention, including formal cardiac rehabilitation and exercise training programs. PMID- 11416344 TI - Renal Considerations in Geriatric Patients With Heart Disease. AB - Renal and electrolyte complications occur commonly in elderly patients with heart disease. Renal function declines with age. A seemingly normal serum creatinine level in the geriatric patient often represents a creatinine clearance of 60 ml/min or less. It is important to measure or estimate the creatinine clearance in an older patient with a borderline high or elevated serum creatinine level before administering renally excreted drugs. The Cockcroft and Gault formula is recommended for estimating the creatinine clearance in such patients. Impaired renal function can also predispose to drug-induced hyperkalemia in geriatric patients; the most common offending drugs are potassium chloride supplements, potassium-sparing diuretics, angiotensin-converting enzyme inhibitors, digoxin, and nonsteroidal anti-inflammatory drugs. Elderly patients should be evaluated for renal artery stenosis if they have worsening of previously stable hypertension, new-onset hypertension, or progressive renal impairment on angiotensin-converting enzyme inhibitors. Risk factors and management guidelines for radiocontrast nephropathy in the elderly are also discussed. PMID- 11416345 TI - Lessons From the Cardiovascular Health Study Question and Answer Format. PMID- 11416346 TI - Images in Geriatric Cardiology - Diseases of the Aorta. PMID- 11416347 TI - Atrial Fibrillation in the Elderly. AB - Atrial fibrillation is a common cardiac arrhythmia affecting up to 5% of people over the age of 65 years. The elderly are less well able to withstand the hemodynamic stress of new-onset and rapid atrial fibrillation. Advancing age is a predisposing factor for atrial fibrillation itself and for its complications. The main adverse outcomes caused by atrial fibrillation include peripheral embolic events, particularly stroke and left ventricular dysfunction. Sinus rhythm is of special importance in the elderly as it may be the main way to reduce the thromboembolic risks of atrial fibrillation. When sinus rhythm cannot be restored and maintained, control of heart rate by pharmacologic or other means has advantage in itself. The long-term benefits of aspirin and warfarin in patients with chronic nonvalvular atrial fibrillation have been demonstrated by recent trials. However, these benefits do not universally extend to patients above the age of 75 years, and thoughtful individualized antithrombotic treatment is needed for the elderly patient with atrial fibrillation. PMID- 11416348 TI - Clinical Spectrum of Congestive Heart Failure in the Elderly Population. AB - One hundred and fifty-two patients with congestive heart failure above 65 years of age were treated at The Mercy Hospital of Pittsburgh in the calendar year 1992, forming 38% of the total number of inpatients with this condition for that year. Hypertension, ischemic heart disease, and valvular diseases were the common etiological factors. Evidence for diastolic dysfunction was seen in 34% of patients by echocardiographic Doppler study. Atrial fibrillation was the most common arrhythmia. Acute or subacute dyspnea was the most common presenting symptom. Response to conventional treatment was excellent but readmissions for recurrence of congestive heart failure were very common. The in-hospital mortality of these patients in that year was 6%. The possible precipitating factors for recurrent admissions are discussed. In order to favorably control precipitating factors and reduce readmissions, a multicenter registry of congestive heart failure in the elderly is suggested. PMID- 11416350 TI - Images in Geriatric Cardiology - Symptoms of Dizziness, Loss of Balance, and Confusion. PMID- 11416349 TI - Non-Q-Wave Myocardial Infarction in the Elderly: Clinical Characteristics and Management. AB - Patients with non-Q-wave myocardial infarction (MI) have been shown to be distinctly different from patients suffering Q-wave MI with respect to pathophysiology, clinical characteristics, and natural history. The management of the increasing group of geriatric patients with non-Q-wave MI imposes several challenges to the practicing cardiologist. This paper tries to elucidate the specifics of non-Q-wave MI and draw conclusions regarding the current understanding and management of elderly patients with non-Q-wave MI based on available data. However, there are a limited number of studies addressing this topic. Further trials are needed in order to improve the current understanding of the most reasonable approach to the growing number of geriatric patients presenting with non-Q-wave MI. PMID- 11416351 TI - Open Heart Surgery in Octogenarians: A Review. AB - Until fairly recently, cardiac surgery was controversial in octogenarians. With the improvement of life quality and health services, the number of octogenarians in the population is steadily increasing. Furthermore, the improvement of surgical techniques and perioperative care permit safer cardiac surgery in this age group. Surgery is beneficial especially for patients undergoing coronary revascularization (early mortality rate, 0% to 12%) or isolated valve surgery (early mortality rate, 1.8% to 20%). Great caution should be exercised when considering candidates for combined coronary and valvular or multiple valve surgery as mortality is much higher in this group. Overall, the operative course in the very elderly is more complicated and is associated with various postoperative complications in 57% to 97% of the patients, which is reflected in longer postoperative hospitalization-an average of 10-19 days. Nevertheless, the 5-year survival in these patients is 47% to 71%. With proper selection of patients, carefully planned surgery, and meticulous postoperative care octogenarians can enjoy prolonged life expectancy and improved quality of life following open heart surgery. PMID- 11416352 TI - Improvement in the Prognosis of Patients With Acute Myocardial Infarction in the 1990s Compared With the Prethrombolytic Era: An Analysis by Age Subgroups. AB - The number of elderly patients experiencing myocardial infarction (MI) is growing rapidly, and their hospital mortality rate remains high, although mortality after acute MI declined in the 1990s. The in-hospital and first-year mortality rates in 5,839 patients with acute MI in 1981-1983 were compared with 1,014 consecutive patients admitted in 1992 to the coronary care units in Israel. The clinical characteristics of gender, prior MI, and acute MI location were similar in the age subgroups in both periods. Patients admitted in 1981-1983 did not receive thrombolytic therapy, percutaneous transluminal coronary angioplasty (PTCA), or coronary artery bypass grafting (CABG), whereas in those admitted in 1992, the frequency of thrombolytic therapy in the age subgroups at or below 55 years, 56 74 years, and at or above 75 years decreased with advanced age (56%, 48%, and 24%, respectively, P is less than.0001). The frequency of PTCA also declined (11%, 7%, and 3%, respectively, P is less than.002). The in-hospital mortality rate for the entire study population declined from 18% in 1981-1983 to 11% in 1992 (adjusted odds ratio [OR] 0.49; 95% confidence interval, 0.39-0.62). The decline in in-hospital mortality was more marked among patients aged 56-74 (18.0% 10.0%; OR 0.48) and those aged at or above 75 years (35.0%-21.5%; OR 0.44), than among patients aged at or below 55 years (6.9%-4.3%; OR 0.70). This decline was most marked in reperfused patients, particularly the very elderly (OR 0.31; 95% CI 0.14-0.69). The cumulative first-year mortality declined by 40% (from 25.5% in 1981-1983 to 17.9% in 1992, hazard ratio (HR) 0.61; 95% CI 0.52-0.72). The decline was more marked among patients aged 56-74 years (26.2%-16.5%; HR 0.56) and those aged at or above 75 years (49.6%-37.6%; HR 0.58) than among counterparts aged at or below 55 years (10.2%-7.9%; HR 0.86). The decline was most marked in reperfused patients, particularly the very elderly (HR 0.31; 95% CI 0.16-0.59). In conclusion, (1) during the last decade in Israel, cumulative first-year mortality after an acute MI declined by 40% and was most marked in patients aged greater than 55 years and in particular the very elderly (at or above 75 years); (2) the main improvement in survival was achieved early after the acute MI, and was maintained thereafter during the first postinfarction year; 3) the favorable outcome in 1992 is related to changes in patients management, mainly reperfusion therapy (thrombolysis, PTCA, CABG); and (4) reperfusion therapy in elderly patients with acute MI should be considered systematically unless specific contraindications are present. PMID- 11416353 TI - Percutaneous Transluminal Coronary Angioplasty in Octogenarians. AB - Revascularization is a theoretically ideal technique for treating repeated angina pectoris in elderly patients. Coronary angioplasty is an attractive option if it can be performed safely. In this report, we describe our experience with 80 angioplastic procedures in 72 patients older than 80 years. Seventy-two of these procedures (90%) were performed after 1990 using the currently available, improved angioplasty equipment. A third of these patients did not have any risk factors for coronary artery disease and 45% had only 1 risk factor. A history of previous myocardial infarction or coronary bypass surgery was present in 48 (67%) patients. In most patients (72%), angioplasty was performed during an admission for unstable angina or acute myocardial infarction. Single-vessel disease was present in 22% of patients. Successful angioplasty was performed in 92% of patients and major complications occurred in 8% of patients. Four patients (6%) died; in all of them, angioplasty was performed emergently to treat cardiogenic shock. None of 68 patients who underwent elective angioplasty for stable angina or a semielective procedure for unstable angina died during the hospitalization. We conclude that angioplasty can be performed safely in the carefully selected octogenarian. There is a low-risk subgroup of patients with limited disease in whom angioplasty is an attractive treatment strategy. PMID- 11416354 TI - Permanent Cardiac Pacing in Octogenarians. AB - A significant percentage of patients in need of a permanent pacemaker are older than 80 years. The implantation policy may be determined either by the patient's physical activity or by chronologic age. The trend in pacemaker implantation in patients over 80 during the last 10 years in our institution was evaluated and compared with the trend in the patients younger than 80 at the time of implantation. Of 519 patients who had primary pacemaker implantation, 152 (29%) were older than 80 at the time of the procedure. Another 189 patients had second implantation procedures, and 80% of them were older than 80 years. Complete atrioventricular block was the indication for pacing in 44 A+/- 11% and sick sinus syndrome in 25 A+/- 7%. The tendency to implant dual-chamber pacemakers increased from 0% during 1985 to 76% in 1994, including 69% DDD and 31% DDDR, but the transition was faster in the younger group. By 1994, there was no difference in the incidence of advanced pacing systems in the 2 age groups. During 1985, only VVI pacemakers were replaced, and during 1994, less than 10% were replaced with simple ventricular pacing units. Pacing system upgrading was frequent during the second half of the decade. The success and complication rate of implantation did not differ in the 2 groups. PMID- 11416356 TI - Epidemiology and Prevention of Cardiovascular Diseases in Elderly People: Report of a WHO Study Group. AB - Reproduced by permission of WHO, from: Epidemiology and Prevention of Cardiovascular Diseases in Elderly People. Report of a WHO Study Group. Geneva, World Health Organization, 1995 (Technical Report Series, No. 853). PMID- 11416355 TI - Rehabilitation in Older Coronary Patients. AB - Modern medicine dictates a more active therapeutic approach in the geriatric population. This review summarizes relevant data on cardiac rehabilitation in patients aged 65 years or older. The cardiovascular risk factors in the elderly are similar to those in younger people; thus, treating hypertension or hypercholesterolemia, cessation of smoking, and changing of life style and proper diet are recommended regardless of age. Cardiovascular function gradually declines with age, and the incidence of atherosclerotic heart disease increases. Rehabilitation programs for elderly patients with postmyocardial infarction should be specifically designed, particularly for exercise training. Special attention should be paid to warm-up and cool-down periods, upgrading the intensity of exercise, and programming the muscular conditioning component. On the whole, cardiac rehabilitation in the elderly is cost-effective. It should consider the individual's general health, risk factor profile, cardiac status, and mental response. PMID- 11416357 TI - Images in Geriatric Cardiology - Worsening Dyspnea and Fatigue not Responsive to Medical Management. PMID- 11416360 TI - Epidemiology and Etiology of Congestive Heart Failure in the Elderly. PMID- 11416359 TI - Normative Aging Changes in Cardiovascular Structure and Function. PMID- 11416361 TI - Heart Failure in the Elderly: Systolic and Diastolic Dysfunction. PMID- 11416362 TI - Signs and Symptoms of Heart Failure in Elderly Patients. PMID- 11416363 TI - The Diagnostic Utility of Noninvasive Tests in the Office or Outpatient Evaluation of the Elderly Patient With Congestive Heart Failure. PMID- 11416365 TI - Diet and Lifestyle Changes: Compliance With Family Counseling. PMID- 11416364 TI - Diagnostic Differentials of Co-Existing Diseases in the Elderly Patient With Heart Disease PMID- 11416366 TI - Age-Related Pharmacodynamic Changes in the Elderly: Treatment Considerations in Heart Failure. PMID- 11416367 TI - Management of Heart Failure Attributable to Ischemic Heart Disease and Other Dilated Cardiomyopathies. PMID- 11416368 TI - Hypertensive Heart Disease, Hypertrophic Cardiomyopathy and Heart Failure in the Elderly. PMID- 11416369 TI - Management of Valve Disease in the Elderly. PMID- 11416370 TI - Arrhythmias in the Elderly: Office Assessment. PMID- 11416371 TI - Exercise Benefits and Prescription in Chronic Heart Failure: The Elderly Patient. PMID- 11416372 TI - Oral Anticoagulant Therapy at Elderly Age: Heart Failure and Nonvalvular Atrial Fibrillation. PMID- 11416373 TI - Concise Guide to the Management of Heart Failure - World Health Organization/Council on Geriatric Cardiology Task Force on Heart Failure Education. PMID- 11416375 TI - Images in Geriatric Cardiology - Misdiagnosis of Coronary Artery Disease. PMID- 11416374 TI - Report From the American Heart Association 68th Annual Scientific Sessions: Focus on Congestive Heart Failure. PMID- 11416377 TI - Rehabilitation of the Elderly Cardiac Patient: Recommendations of the Clinical Practice Guideline Cardiac Rehabilitation. PMID- 11416376 TI - Detection of Progression of Coronary Artery Disease in the Elderly. AB - In order to assess the progression of coronary artery disease (CAD) in the elderly, we evaluated 91 patients aged 75 years or older who had undergone 2 consecutive angiograms without intervening revascularization. Progression was defined as an absolute increase in lumen narrowing by at least 20% with minimum stenosis of 50% at second angiogram, or progression to total occlusion of any preexisting lesion. Progression involving at least 1 vessel was observed in 63% of patients. Only 6% of initially normal or insignificantly diseased segments showed progression. In contrast, 72% of segments that progressed to total occlusion had shown an initial narrowing greater than 75%. Progression occurred in 100% of patients with an interval myocardial infarction, but in no patients with symptomatic valvular disease. No regression was observed, while 7 of 36 (19%) of initially occluded segments had recanalized. We conclude that progression of CAD in the elderly occurs at rates similar to those observed in a younger population; however no correlation could be found between the rate of progression and either risk factors or elapsed time between angiographic studies. PMID- 11416378 TI - Repetitive Hospital Admissions for Congestive Heart Failure in the Elderly. PMID- 11416379 TI - Images in Geriatric Cardiology - A Rare Case of Giant Left Atrium. PMID- 11416380 TI - Images in Geriatric Cardiology - Fever, Anorexia, and Malaise in a Patient Awaiting Hip Arthroplasty. PMID- 11416381 TI - Heart Disease in the Elderly: A Joint Symposium - Co-Sponsored by the Japan Geriatrics Society and the Council on Geriatric Cardiology. PMID- 11416382 TI - Hospitalization and Health Care Costs Due to Congestive Heart Failure in the Elderly. AB - The objective of this study is to describe the occurrence of readmissions due to congestive heart failure (CHF) in an elderly population and to give insight into the costs associated with readmissions. Two hundred and seven patients ages 65 84, hospitalized due to CHF, were included in the analysis. Seventy-six patients (37%) were readmitted due to CHF within 1 year including 32 (16 %) readmitted within 30 days. The patients with early readmissions had a significantly shorter index hospitalization compared to those not readmitted. The total cost for hospitalization due to CHF was 7,739,488 SEK (8 SEK to $1US). Readmissions accounted for 41% of this amount. We conclude that readmissions are common in patients with CHF and that the frequencyof early readmissions for CHF may be related to the duration of prior hospitalization. Early discharge may lead to increased health care expenditure due to a high frequency of readmissions. PMID- 11416383 TI - Elderly Cardiac Rehabilitation Patients Show Greater Improvements in Ventilation at Submaximal Levels of Exercise. AB - The minute ventilation (VE) and carbon dioxide output (VCO&sub2;) adaptations to 12 weeks of cardiac rehabilitation were investigated in diagnosis-matched elderly patients (ELD) and younger patients (YNG). Thirty ELD (25 males, 5 females; age equals 69.2A+/-3.9 years) and 30 YNG (27 males, 3 females; age equals 48.3A+/-7.0 years) were exercise tested at program entry (T1) and completion (T2). Peak oxygen uptake (VO&sub2; peak) and ventilatory threshold (VT) (ml kg-1 min-1) were greater for YNG at both T1 and T2 (p is less than 0.01). Both ELD and YNG showed significant T1 to T2 improvements (p is less than 0.01) for VO&sub2; peak (21.1% and 23.4%) and VT (20.1% and 12.4%). The VE/VCO&sub2; ratio was greater (p is less than 0.01) for ELD but approached YNG values after training. The VCO&sub2;/VO&sub2; slope showed a greater T1 to T2 reduction in ELD (12.4% vs 3.4%). There was no relationship (r(2) equals 0.15; NS) between the percentage change in VO&sub2; peak and VE/VCO&sub2; indicating unrelated mechanisms of improvement. We conclude that ELD showed similar improvements in VO&sub2; peak but greater improvements in their ventilatory tolerance for submaximal exercise. These changes indicate improvements in aging-enhanced exercise ventilation, are important for performance of normal activities of daily living, and would have been undetectable without gas exchange analysis. PMID- 11416384 TI - Isolated CABG in the Elderly: Operative Results and Risk Factors Over the Past Three Decades. AB - This chapter is reproduced from the book, Coronary Bypass Surgery in the Elderly, edited by Paul J. Walter. Reprinted by permission of Kluwer Academic Publishers. All Rights Reserved (c)1995 Kluwer Academic Publishers. PMID- 11416385 TI - Images in Geriatric Cardiology - Ventricular Septal Perforation by Pacemaker Lead After Cardiac Defibrillation. PMID- 11416386 TI - Images in Geriatric Cardiology. PMID- 11416387 TI - Update on Hyperlipidemia in the Elderly: Is This a Risk Factor for Heart Disease? AB - Results concerning the utility of lipid screening in the elderly are conflicting. Many studies have shown no association between total cholesterol measurements in the elderly and the development of coronary heart disease (CHD). Several recent investigations, with a few exceptions, have demonstrated that high-density lipoprotein-cholesterol (HDL-C) levels and the total/HDL-C ratio are, however, effective markers for CHD in older populations. Although the relative risk associated with lipid measurements tends to decline in older persons, the attributable risk for CHD associated with higher cholesterol and lower HDL-C tends to be greater in older persons because of a greater number of events in this population. Clinical trials with modern lipid-lowering agents have demonstrated efficacy and safety in middle-aged subjects with the newer medications, and it is time to consider a lipid altering clinical trial specifically targeted to persons over the age of 65 years to determine whether morbidity and mortality can be reduced. PMID- 11416388 TI - Drug Therapy of Lipid Disorders: Benefits and Limitations in Elderly Patients. AB - Pharmacologic therapy of lipid disorders has become an established means to reduce cardiovascular morbidity and mortality. Compared to middle-aged individuals with hyperlipidemia, the use of such treatment in the elderly population has been controversial, as there is only scant data from large clinical trials that can be applied to this group. Therefore, current recommendations for specific anti-lipid drug use in older individuals must weigh each drug's potential benefits against the potential treatment risks that are inherent to this age group. PMID- 11416389 TI - The Effect of Exercise Training on Serum Lipids in the Elderly. AB - Numerous intervention trials in young subjects suggest that aerobic exercise training exerts favorable effects on specific lipid subfractions, in particular serum triglycerides and high density lipoprotein (HDL) cholesterol. Cross sectional studies in older individuals suggest that active individuals have more favorable lipid profiles than inactive individuals. However, it remains controversial as to whether leisure time activity or fitness levels per se are independent predictors of lipid measures in the elderly versus their intermediate effects on body composition and body fat distribution. That is, a greater deposition of abdominal fat in the least fit individuals is associated with adverse lipid profiles. Short-term (8-weeks) aerobic exercise in elders is not associated with any significant changes in lipid profiles, however, longer term programs, particularly if associated with improvements in body fat distribution and/or weight loss, have been demonstrated to yield significant improvements in lipid subfractions, in particular triglycerides and HDL cholesterol. PMID- 11416390 TI - Results of Lipid-Lowering Trials in the Elderly and a Look to the Future. AB - A lack of clinical research in the elderly hampers the creation of specific therapeutic regimens to treat dyslipidemia in this group. However, extrapolation of data from studies in middle-aged men may be appropriate since the mechanisms of atherogenesis do not appear to change significantly with advancing age. The results of the secondary prevention Scandinavian Simvastatin Survival Study (in which 52% of the patient population was 60 years of age or older) and the primary prevention West of Scotland Coronary Prevention Study (in which the mean age of the patient population was older than those of other primary prevention trials) suggest the benefit of lipid-lowering in older populations. More clinical trials that include elderly subjects are needed to evaluate fully the efficacy of lipid regulating therapy in older patients. PMID- 11416391 TI - The Social, Ethical, Economic, and Political Implications of Coronary Heart Disease Prevention in the Elderly. AB - A variety of health measures during this century has resulted in marked increases in the number of people reaching old age, particularly 85 years of age or more. Two recent measures include lifestyle modification to lessen rates of coronary heart disease (CHD) and control of hypertension to decrease stroke events as well as to lessen CHD risk. This review presents approaches for considering the social, ethical, economic, and political implications of prevention of CHD in the elderly. It reviews demographic and other characteristics of our aging society and discusses what thier implications are as research in the biology of aging results in the prolongation of life. PMID- 11416392 TI - Are the Hemodynamic Changes in Elderly Hypertensive Patients of Clinical Importance? Do They Influence the Choice of Medication? AB - Despite the fact that elderly hypertensive patients have numerous concomitant diseases and changes in physiologic responses compared to middle-aged or younger individuals, they generally repond well to antihypertensive drug therapy without significant adverse reactions-provided that medication is started at low dosages and that dosages are increased slowly. There is little doubt that the benefit of treatment outweighs the risk. PMID- 11416393 TI - Hypertension in the Elderly: Lifestyle Modification. AB - Five modalities of nonpharmacologic approaches are recommended at present for lifestyle modification and control of arterial blood pressure elevation: 1) weight reduction to ideal body weight, since it reduces risk of hypertension as well as overall cardiovascular morbidity and mortality; 2) dietary sodium restriction to less than 2 g a day, without assurance that it will normalize arterial pressure although it may help reduce dosage and numbers of prescribed antihypertensive drugs; 3) moderation of alcohol consumption to less than 1 ounce a day; 4) a regular isotonic exercise program; and 5) cessation of tobacco consumption. Other nonpharmacologic approaches have yet to be shown to be effective in controlling arterial pressure. Adherence to these measures has probably contributed to a reduction in the number of patients with hypertension in the United States from 59 to 50 million. PMID- 11416394 TI - Treatment of Elderly Hypertensive Patients With Diabetes, Renal Disease, and Coronary Heart Disease. AB - Aging in Westernized industrialized societies is associated with an increasing prevalence of hypertension, type II diabetes mellitus, renal disease, and atherosclerotic vascular disease. This increase in the chronic disease processes in industrialized societies is related, in part, to increasing obesity, reduced physical activity, medications such as nonsteroidal anti-inflammatory agents, and other environmental influences. Hypertension in the elderly is characterized by high peripheral vascular resistance, reduced baroreflex sensitivity, a low renin state with reduced cardiac output/increased hypertrophy, reduced intravascular volume, and an increased propensity to salt-sensitivity. Initial antihypertensive therapy in the elderly patient should be based on attempts to affect hygienic measures such as weight reduction, decreased salt and fat intake, and a careful aerobic exercise program. The initial antihypertensive drugs of choice are low doses of diuretics, which have been shown to reduce cardiovascular mortality in the elderly. Low doses of diuretics do not substantively affect carbohydrate and lipid metabolism. Lipid abnormalities in the elderly should generally be treated in a similar fashion to those in the middle-aged individual. Compliance with medical therapy in the elderly patient has been demonstrated to be relatively good. PMID- 11416395 TI - Hormone Replacement Therapy in Postmenopausal Women: Does It Lessen Coronary Heart Disease Risk? AB - Coronary heart disease is the leading cause of death in adult women in the U.S. Since clinical manifestations of coronary disease predominate in postmenopausal women, the need to evaluate coronary disease-gonadal hormone relationships is compelling. Further, estrogen exerts biologically plausible benefits: improved lipid profile, lower fibrinogen levels, favorable changes in vascular reactivity, possible antioxidant effects, among others. Although observational studies of estrogen use suggest substantial lessening of coronary risk, selection biases favoring a healthy cohort of women mandate randomized clinical trial data, with such trials currently in progress. Information is lacking regarding benefit: risk ratios of transdermal estrogen and estrogen/progestin combinations as coronary preventive therapies. PMID- 11416396 TI - Images in Geriatric Cardiology - A Large Myxoma Originating at the Foramen Ovale and Presenting as Congestive Heart Failure. PMID- 11416397 TI - The Intersecting Paths of Geriatrics and Cardiology. PMID- 11416398 TI - Angiotensin II-Receptor Blockers: Profile of a New Drug Class for Antihypertensive Therapy. AB - Of the population in the United States 65 years of age or older, more than half have high blood pressure. The benefits of treatment are well established, with decreased morbidity and mortality from stroke and MI. Antihypertensive therapy available for the elderly now includes losartan, the prototype of a new class of drugs, the angiotensin II-receptor blockers. The likelihood of even fewer side effects with the angiotensin II-receptor blockers than with ACE inhibitors promises to make them an important new therapy for elderly patients. Angiotensin II is the primary vasoactive component of the renin-angiotensin system, which is implicated in the pathophysiology of hypertension. Influencing this system pharmacologically has, therefore, important consequences for high blood pressure treatment. The ACE inhibitors have had success as antihypertensive agents, but they act early in the renin-angiotensin cascade, possibly leading to unwanted effects. The angiotensin II-receptor blockers act directly at the receptor level, thus avoiding the complications of interrupting the metabolic pathway early and disrupting other hormonal systems. These agents represent a new treatment option for hypertension in the elderly. Other clinical indications, such as heart failure and post-MI, are being investigated. PMID- 11416399 TI - Influence of Old Age, Gender, and Internal Mammary Artery Grafting on Operative Mortality and Morbidity in Coronary Artery Bypass Grafting. AB - Operative risk may change its pattern from time to time. To investigate determinants for operative mortality in patients undergoing CABG since the mid 1980s and the influence of age, gender, and IMAG on mortality, the data of 6,360 patients who underwent CABG from January 1986 through June 1993 were analyzed. Of these patients, 2,153 had SVG alone and 4,207 had IMAG including UIMAG (3,957) and BIMAG (250). Overall mortality was 4.34%. OM (in hospital death) for IMAG (2.69%) was lower than for SVG (7.57%, p is less than 0.0001). There was no difference in mortality between BIMAG (3.2%) and UIMAG patients (2.65%, p equals 0.6) or UIMAG/SVG patients (4.29%, p equals 0.36). Fewer IMAG patients had postoperative complications (LCO, insertion of IABP, prolonged ventilation, reoperation for bleeding, neurological complications, perioperative MI, and infection of legs) than SVG patients. There was no difference in the incidence of sternal infection. To determine risk factors for mortality and the influence of IMAG on the outcome, 82 variables (31 preoperative, 17 intraoperative, and 34 postoperative) were analyzed by univariate analysis. Significant variables or the variables having a trend (p is less than 0.2) to be associated with mortality were included in stepwise multiple logistic regression analyses. Two regression analyses were separately performed. Regression 1 only included pre- and intraoperative variables whereas regression 2 included postoperative variables as well. The logistic regressions demonstrate that preoperative (low EF, age at or above 70, female gender, history of CHF or arrhythmia, and functional Class), intraoperative (emergency operation, reoperation, long perfusion time, and lack of IMAG), and postoperative (complications) variables are independently associated with higher mortality. Female gender is an independent determinant for mortality and not dependent on small body surface area. Neither use of BIMAG or right IMAG, nor number of grafts is associated with the OM. The identification of these risk factors may have important implications in further improvement of the results for CABG. PMID- 11416400 TI - Elderly Patients With no Prior Cardiopulmonary Disease Show Ventilation/Perfusion Lung Scan Characteristics That Are Sensitive and Specific for Pulmonary Embolism. AB - The purpose of the present investigation was to test the hypothesis that stratification according to age, in addition to stratification according to the presence or absence of prior CPD, in elderly patients may show criteria on V/Q lung scans that have a higher sensitivity for acute PE than the entire population, while maintaining a high specificity and high PPV. Data from the PIOPED were evaluated among 1,050 patients with suspected acute PE in whom the diagnosis was made or excluded by pulmonary angiography. Age groups were at or above 70 years of age (n equals 259), 40-69 years of age (n equals 585) and 18-39 years of age (n equals 206). Patients were stratified according to the presence or absence of prior CPD. Lung scans were evaluated on the basis of the cumulative number of mismatched vascular perfusion defects (large or moderate size mismatched segmental perfusion defects). Among patients at or above 70 years of age who had no prior CPD, greater than or equal to 2 matched vascular perfusion defects showed a sensitivity of 23 of 31 (74%), a specificity of 29 of 29 (100%), and a PPV of 23 of 23 (100%). Stratification according to both age and CPD was particularly useful for the evaluation of V/Q lung scans in patients at or above 70 years of age with no prior CPD, although only 23% of patients at or above 70 years of age had no prior CPD. PMID- 11416401 TI - Opportunities to Improve the Cost-Effectiveness of CABG Surgery. AB - This chapter is reproduced with permission from the book, Coronary Bypass Surgery in the Elderly: Ethical, Economical and Quality of Life Aspects, edited by Paul J. Walter, and published by Kluwer Academic Publishers. PMID- 11416402 TI - Images in Geriatric Cardiology - Fever and Chills Two Months Post Pacemaker Placement. PMID- 11416403 TI - Myocardial Revascularization in Patients 70 Years of Age and Older Without the Use of Extracorporeal Circulation. AB - Myocardial revascularization in elderly patients (i.e., over 70 years of age) is associated with higher incidence of morbidity and mortality compared with younger patients. We herein report our experience on myocardial revascularization in the elderly comparing the results between 2 groups; one operated on with the aid of cardiopulmonary bypass and the other group in whom extracorporeal circulation was not used. The records of 265 elderly patients ( at or above 70 yrs) undergoing myocardial revascularization were prospectively analyzed between January 1994 and December 1995. Mean age was 74 A+/- 6 years (range 70-95 yrs), with 83 (31.3%) females and 182 (68.7%) males. The following were the preoperative diagnoses: chronic ischemia and angina (186 patients), reoperation (28 patients), unstable angina (26 patients), failed angioplasty (13 patients), post-thrombolytic therapy (7 patients), cardiogenic shock (2 patients), evolving myocardial infarction (2 patients), and aortic dissection (1 patient). Extracorporeal circulation was used in 204 (76.9%) patients (Group I) and no extracorporeal circulation was used in 61 (23.1%) patients (Group II). The overall mortality was 6.4%, with 7.8% (16/204) in Group I and 1.6% (1/61) in Group II. Hospital stay was 11.4 days in Group I and 7.1 in Group II. Transfusion requirements were 1.4 and 0.6 units for Groups I and II, respectively. The use of extracorporeal circulation in the elderly is a major cause of morbidity and mortality following myocardial revascularization. Whenever possible, myocardial revascularization in the elderly should be performed without the use of extracorporeal circulation. PMID- 11416404 TI - Pacemakers in the Elderly: Selection of Pacing Systems. AB - Abnormalities in impulse formation and conduction are common in the elderly. Changing technology has led to new pacemakers of enormous complexity and flexibility. Hemodynamic changes due to aging include a greater contribution of atrial contraction to ventricular filling. Dual-chamber pacing which maintains atrial transport, is associated with less incidence of chronic atrial fibrillation, stroke, heart failure, pacemaker syndrome, and death. Bipolar systems are free of inhibition due to pectoral myopotentials and cross talk in the case of dual-chamber pacing. Invasive electrophysiologic studies have very low risk of complications and can provide important information at the time of choosing the optimal pacing system for a specific patient. In the patient with stable atrial rhythm, pacing systems that maintain atrial transport are preferred over simple ventricular pacing. PMID- 11416405 TI - Aortic Stenosis in the Elderly: A Decade of Changing Concepts. AB - The last 10 years have been a time of dynamic evolution in the diagnosis and treatment of aortic stenosis in elderly patients. The pathology of diseased aortic valves has been better defined. Improvements in cardiac catheterization and Doppler echocardiographic assessment have been introduced. The initial promise of balloon valvuloplasty and ultrasonic valvular debridement was short lived. Operative results improved and the importance of associated coronary disease has been investigated. While many issues remain unresolved, the lessons of the last decade should improve our clinical success in the future. PMID- 11416406 TI - Hypertension in the Elderly: Why, When, and How to Treat? AB - Hypertension is accepted as a risk factor for cardiovascular disease in the elderly population. Data supporting the benefit of treating hypertension in the elderly is extensive but it continues to be poorly recognized and treated in the general population. The correct diagnosis is the initial step for avoiding common side effects of the treatment. In this article we summarize important particularities of the diagnosis and treatment of hypertension in the elderly. The decision on when and how to treat hypertension must be carefully evaluated and individualized. Emphasis should be on the patient, treating the elderly patient with hypertension rather than the hypertension in the elderly patient. PMID- 11416407 TI - Atrial Fibrillation at Elderly Age: The Importance of Stroke Prevention. AB - Atrial fibrillation increases in prevalence with advancing age and is the major precursor of embolic stroke in an elderly population. Currently about 2.2 million people in the United States have atrial fibrillation and their median age is 75 years. Patients older than 75 years of age with atrial fibrillation and those with underlying coronary heart disease or with heart failure are at highest risk of stroke. Long-term warfarin anticoagulant therapy, in a number of randomized controlled trials of its use in nonvalvular atrial fibrillation, substantially decreased stroke risk, with benefit maintained at elderly age. Warfarin anticoagulation at elderly age, with careful monitoring and attention to concomitant therapies, can be safely accomplished with an acceptably low rate of bleeding complication. PMID- 11416408 TI - A First-Line Treatment Option for Elderly Hypertensive Patients: A Low-Dose Multimechanism A -Blocker/Diuretic. AB - The high incidence of hypertension among the elderly makes a once-daily multimechanism antihypertensive an advantageous agent for the control of hypertension. Because of its safety and efficacy in earlier studies, a low-dose multimechanism A -blocker/diuretic, bisoprolol fumarate 2.5, 5, or 10 mg combined with HCTZ 6.25 mg, was evaluated in multicenter studies, including a subgroup analysis of elderly patients at or above 60 years of age. Results indicate that all 3 dosage strengths of this once-daily agent are effective in reducing blood pressure in this population throughout a 24-hour period. Since low doses of both components are used to achieve efficacy, the incidence of side effects with all 3 dosage strengths is similar to that of placebo. In particular, this agent does not cause hypokalemia. Bisoprolol fumarate/HCTZ is an appropriate therapeutic option for elderly patients with hypertension because of its ability to effectively and safely control blood pressure in this difficult to treat population. PMID- 11416409 TI - Images in Geriatric Cardiology - Chest Pain Syndrome in Hypertensive Husband and Wife. PMID- 11416410 TI - Management of Peripheral Arterial Disease in the Elderly: Case Report and Review of the Literature. PMID- 11416411 TI - Coronary Artery Bypass Surgery and Health Related Quality of Life: Data From the National Health and Nutrition Examination Survey. AB - This chapter is reproduced with permission from the book, Coronary Bypass Surgery in the Elderly: Ethical, Economical and Quality of Life Aspects, edited by Paul J. Walter, and published by Kluwer Academic Publishers. For further information on the book format see the advertisement appearing on page 37 of the July/August 1996 issue of AJGC. PMID- 11416412 TI - Images in Geriatric Cardiology - Acute Myelogenous Leukemia Presenting as Embolic Marantic Endocarditis. PMID- 11416413 TI - Cardiovascular Disease in the Elderly. PMID- 11416414 TI - Hypertension In The Elderly. AB - Cardiovascular disease is still the most common cause of morbidity and mortality in the elderly and hypertension is a significant risk factor for CVD such as stroke, myocardial infarction, and chronic renal disease. Fortunately, hypertension is readily treatable. A clear benefit from treating elevated blood pressure has been demonstrated by several clinical trials. In fact, benefit from treatment of either systolic or diastolic hypertension in this group is much greater than in younger hypertensives. Understanding physiologic and socioeconomic changes with age is an essential part of treatment and will allow for individualized treatment suitable for the elderly. Life-syle modification is a good starting point to control high blood pressure but should not be enforced unrealistically in this group. Once pharmacologic therapy is decided upon it should be started with a lower dose than usual and should be slowly titrated. Monotherapies and combination therapies are available for treatment of hypertension. All clinicians who take care of the geriatric population should be aware of the benefit of antihypertensive therapy since this is one of the most rewarding aspects of preventive medicine. PMID- 11416415 TI - Exercise and the Health of the Elderly. AB - Most lifetime morbidity is concentrated in the later years as a consequence of increasing amounts of chronic illness and senescent changes. An ideal society would reduce this large illness burden by focusing upon increasing the incident age of first infirmity and compressing lifetime morbidity between that point and the average age at death; the "Compression of Morbidity." We compared progression of musculoskeletal disability, pain, and medical care costs by a longitudinal study of 537 members of a runners club and 423 community controls over a period of more than 8 years with an average initial age of 59 years, with extensive controls for selection bias. Exercising subjects developed disability at a rate only one-fourth that of the sedentary controls. Musculoskeletal pain was reduced by 20%. Medical care costs of exercisers were 25% less than controls. Mortality was significantly reduced in the exercising groups. These findings have substantial implications for health policies directed at increasing amounts of regular lifetime physical activity to improve the quality of life of our rapidly increasing senior population. PMID- 11416416 TI - Hypercholesterolemia in the Elderly: Evaluation and Treatment. AB - Hypercholesterolemia has been established as an important risk factor for atherosclerotic vascular disease. Treatment of elevated cholesterol levels has been shown to clearly reduce the risk of coronary artery events in patients with prior coronary disease as well as with patients without a history of myocardial ischemia. However, in the elderly the association is less clearly defined and the benefits of treating elevated cholesterol levels less established. The following article will discuss the role of hypercholesterolemia in the pathogenesis of coronary artery disease in the elderly and will review the benefits and risks of therapy. PMID- 11416418 TI - Images in Geriatric Cardiology - Complicated Myocardial Infarction. PMID- 11416417 TI - Hypertension in the Elderly: An Update on Treatment Results and Recommendations - A Position Paper From the Council on Geriatric Cardiology. PMID- 11416419 TI - Can Extended-Release Isosorbide Mononitrate be Used as Adjunctive Therapy for Systolic Hypertension? An Open Study Employing Pulse-Wave Analysis to Determine Effects of Antihypertensive Therapy. AB - Augmentation of the systolic ejection pressure wave by an exaggerated reflectance wave, demonstrable by applanation tonometry of the radial artery, is an important contribution to systolic hypertension of the elderly. Therefore, a study of extended-release isosorbide mononitrate, which may reduce the augmentation, was performed in a group of patients aged 49 to 80 years with systolic hypertension. This agent was added, in a dose of 30 to 120 mg once daily, to existing antihypertensive medications and the response of the arterial pulse wave pattern was observed after 7 to 34 weeks of continuing treatment. There were decreases in brachial systolic blood pressure, indirectly measured aortic systolic blood pressure, and pulse wave augmentation. These findings indicate the need for controlled studies of nitrate therapy as an adjunct to treatment with conventional antihypertensive drugs in systolic hypertension of the elderly. The studies should include analysis of pulse waveform to complement conventional sphygmomanometry. PMID- 11416420 TI - Diagnosis and Treatment of Isolated Systolic Hypertension in the Elderly: Results of a Survey Four Years Post-SHEP. AB - There is limited data evaluating the impact on clinical practice of the 1991 SHEP study. To assess present approaches and attitudes to ISH, we surveyed 135 physicians during the spring of 1995. A questionnaire was designed to assess the physician's definition of ISH, views on clinical importance and etiology of ISH, choice of pharmacological intervention (if any), and opinions regarding SHEP's influence on current approaches to the management of ISH. Surveys were distributed to physicians at the weekly Internal Medicine and Cardiology Grand Rounds at The Mt. Sinai Medical Center, New York, NY from February through April 1995. Data were analyzed via Lotus 1-2-3 spreadsheet (Release 3.1 Que Corp.) and responses to opinion statements were factor analyzed on Systat Version 5.0 software. The response rate was 63.7% (87 physician responses). Nearly 50% of the respondents had read the SHEP article and 82.6% had "heard of the study." Approximately 60% believed ISH should be defined in accordance with the SHEP guideline (SBP is greater than 160 mm Hg and DBP is less than 90 mm Hg). Thirty percent of physicians would initiate pharmacological treatment at a SBP less than or equal to 155 mm Hg for patients aged 65-74 years. Of the 85% of physicians (n equals 73) who opted to medicate, the patient's age strongly determined the SBP at which pharmacological treatment would be initiated. Whereas 66% of physicians would use drug therapy for patients aged 65-74 with a SBP less than or equal to 160 mm Hg, 54% and 45% of physicians would consider the same for patients aged 75 84 and 85+, respectively. Thirty eight percent of physicians chose thiazide diuretics as sole first-line therapy. CCB and ACE inhibitors were chosen by 26.8% and 19.7% of physicians, respectively. When compared to younger physicians (less than 60 yrs), older clinicians ( at or above 60 yrs) were more likely to agree that the detection of ISH was not important and that treatment of ISH is ineffective. Survey results demonstrate a definite consensus for initiation of pharmacological treatment in elderly patients with ISH. Of note, a significant percentage of physicians would initiate therapy at SBP less than or equal to 155 mm Hg. This is a level of pressure for which no epidemiological data exists to support treatment. With respect to specific pharmacological treatment of ISH, a clear consensus is still lacking. The increased use of ACE inhibitors and CCB compared with previous studies may have significant impact on the future of treatment costs for the elderly. PMID- 11416421 TI - Age and Gender Related Trends in Body Composition, Lipids, and Exercise Capacity During Cardiac Rehabilitation. AB - Age (40s, 50s, 60s, and 70s) and gender (125 females; 456 males) related trends for peak exercise capacity VO2peak, body composition, and lipids were reviewed for 581 cardiac rehabilitation patients. Groups were equivalent with regard to diagnoses, medications, and risk factors. Peak exercise capacity decreased with increasing age but the percentage improvement attributable to cardiac rehabilitation was similar across each age group (mean of 28.0%). The 70s group had the lowest weights (73.4 kg), similar %fat (31.0%), and the lowest LBM (50.4 kg). They also had lower triglyceride values (191 mg/dL), but the greatest degree of improvement (-23.3%). The percentage improvement for body composition and lipids was similar for the 70s, 60s, and 50s while the 40s group showed no change in weight, %fat, or LBM. Women had lower peak exercise capacities than men but similar relative improvements (26.6% and 28.8%, respectively) as a result of cardiac rehabilitation. Men had greater body weights (85.5 vs. 73.5 kg) and LBM (59.9 vs. 46.9 kg) while women had greater %fat (35.4% vs. 29.3%). Women had a greater percentage weight loss (-1.2% vs. 0.5%) but men had a greater increase in LBM (0.7% vs. 0.4%). Females tended to have higher cholesterol (220 vs. 203 mg/dL) and lower triglycerides (196 vs. 223 mg/dL) but had greater relative improvements in both when compared to men (-4.8% vs. -1.6%; -12.9% vs. -8.2%, respectively). Cardiac rehabilitation has favorable outcomes with regard to peak exercise capacity, body composition, and lipids, across each age group regardless of gender. There is a need for greater emphasis on body composition improvement for females (particularly decreases in %fat and increases in LBM), and lipid reduction (particularly in the younger age groups). PMID- 11416422 TI - Images in Geriatric Cardiology - Functional Single-Leaflet Mitral Valve. PMID- 11416423 TI - Electrocardiology Teacher Analysis and Review - Interatrial Block. PMID- 11416424 TI - Living Not Only Longer, But Better. PMID- 11416427 TI - Public Health Aspects of Out-of-Hospital Sudden Cardiac Arrest Among Elderly African-Americans. AB - In the U.S., the ethnic minority populations of elderly Americans are growing faster than the elderly population as a whole. It has been estimated that by the year 2030, minority elders will make up one-fourth of all elderly Americans. Among minority populations, African-Americans are at the highest risk for CHD and SCA. Approximately two-thirds of all out-of-hospital SCA cases are 65 years of age and older. The rate of a bystander initiated CPR as well as the rate of survival after out-of-hospital SCA is significantly lower among blacks than among whites. Studies have shown that racial differences in outcomes of out-of-hospital SCA cannot be fully explained by features of collapse or relevant service factors and that the underlying socioeconomic and health status of the minority population are factors likely to affect outcome of SCA. Most minority populations in the U. S. have a poorer socioeconomic and health status and access to the health care system than their white counterparts. SCA, as a leading cause of mortality, has been found to be associated with socioeconomic status and has been identified as a key area in which to study minority populations. This brief review summarizes public health aspects of SCA among elderly African-Americans with suggestions for new research and preventive interventions needs. PMID- 11416428 TI - Stroke in Elderly African-Americans. AB - About 85% of strokes in the U.S. occur in people over 65 years of age. As such, stroke is the third leading cause of death, and stroke management and prevention are important topics in geriatric cardiology. Approximately 50%-80% of strokes are preventable. Therefore, treatable stroke risk factors such as hypertension, atrial fibrillation and other manifestations of heart disease, cigarette smoking, diabetes mellitus, excessive alcohol intake, TIAs, and hypercholesterolemia must be identified and vigorously controlled. Once TIAs or other signs of stroke appear, the patient should be immediately transported to the nearest hospital emergency room arriving within one or two hours following the onset of symptoms. Currently accepted approaches to the management of acute stroke symptoms ("Brain Attack") are reviewed. New therapeutic and preventive stroke measures may help reverse the recent national rise in stroke occurrence and mortality as seen most prominently in African-Americans. PMID- 11416430 TI - Electrocardiology Teacher Analysis and Review -Diagnostic ECG in Pulmonary Emphysema. PMID- 11416429 TI - Heart Failure in Elderly African-Americans. AB - Heart failure has emerged as one of the most important cardiovascular diseases in the elderly and has a major functional, economic, and social impact in older individuals. Racial differences in the morbidity and mortality, as well as in rates of hospitalization, have been reported in national surveys and epidemiologic studies. Because of the higher prevalence of hypertension and left ventricular hypertrophy in African-Americans compared to whites, these risk factors may be the principle determinants of the hemodynamic and clinical features of heart failure in African-Americans. The contribution of coronary disease with left ventricular remodeling secondary to myocardial ischemia or infarction varies among African-American populations, depending on the prevalence and impact of traditional risk factors for coronary heart disease. Atherosclerosis may play a greater role in the pathogenesis of heart failure in African-Americans as the impact of urbanization and the adoption of unhealthy lifestyles results in increased prevalence of risk factors for coronary artery disease. The hemodynamic and functional features of ventricular dysfunction in African-Americans with heart failure may range from asymptomatic diastolic dysfunction to typical systolic dysfunction. The differentiation of diastolic from systolic dysfunction, and hypertensive from ischemic cardiomyopathy, may be difficult in African-Americans and usually requires utilization of noninvasive cardiac diagnostic techniques. In addition to the type of ventricular dysfunction and the role of neurohormonal activation, consideration of age-related differences, ventricular structure and function, and variations in drug metabolism and pharmacokinetics are important. PMID- 11416431 TI - Images in Geriatric Cardiology - Myocardial Infarction by Enzymes With New Atrial Fibrillation. PMID- 11416433 TI - Hypertension in Elderly African-Americans. AB - Epidemiologic observations clearly demonstrate that African-Americans present with an earlier onset of hypertension, and experience higher rates of morbidity and mortality associated with elevated levels of blood pressure. Racial differences in blood pressure magnitude begin earlier in life in African Americans, and vascular damage and target organ injury occurs more rapidly. Consequently, elderly hypertensive African-Americans are at significant risk for vascular disease and target organ injury and require aggressive management efforts. Primary prevention of hypertensive vascular disease must focus on intensive risk factor intervention such as the prevention of obesity, modification of caloric, salt, and saturated fat intake, and, if necessary, control of diabetes. Additionally, efforts need to be placed on early identification and education, particularly in areas of socioeconomic disadvantage. PMID- 11416434 TI - The Epidemiology of Coronary Heart Disease and Stroke in African-American Elders. AB - To assess the magnitude of the problem of heart disease and stroke in African American elders, data were reviewed from several national surveys conducted by the National Center for Health Statistics. With the exception of persons aged 75 and over, blacks had higher death rates than whites. Since 1979, heart disease death rates have continued the long term decline in each age, sex, and race group; however, stroke death rates have nearly ceased declining after 1992. In 1990 through 1992 there were probably over 2 million black persons with self reported heart disease and 400,000 with self-reported stroke in the civilian noninstitutionalized population in the U.S. In 1993, 354,000 black persons were discharged from short stay hospitals with a principal diagnosis of heart disease and 97,000 with stroke. Vigorous efforts are needed toward primary prevention of heart disease and stroke to reduce the burden of illness. PMID- 11416435 TI - Access to Care for Elderly Persons. AB - Access to appropriate care for elderly persons for cardiovascular diseases is still a major problem for some segments of the population and some geographic regions. The problem of access can be defined in terms of three principle components: 1) lack of personal resources (insurance); 2) inadequate facilities (hospitals, clinics and medical personnel; and 3) inappropriate withholding of care (principally due to ethnic or culturally-based biases). Solutions to these defined problem components can be planned along the same lines and will require pooling of federal, state, and local government resources; education of the lay public; improved professional education; and raising the level of tolerance for substandard practices based upon peer review and the promulgation of standards of practice via guidelines. PMID- 11416436 TI - Left Ventricular Hypertrophy in the Elderly. AB - The prevalence of LVH increases markedly with aging approaching one third of men and one half of women 70 years or older. Several determinants of LVH have been defined including systemic hypertension, obesity and several other hemodynamic and nonhemodynamic stimuli. The grave prognostic role of LVH has been well documented and the following potential sequelae of LVH have been identified: myocardial ischemia and infarction, ventricular arrhythmias, heart failure with a preserved or impaired left ventricular systolic function, cerebrovascular disease and cardiovascular mortality. The presence of comorbid conditions coupled with the high prevalence rate of LVH in the elderly population enhances the susceptibility of this population and underscores the urgent need to adopt strategies that help to prevent the development of LVH and explore the potential beneficial effects of reversing it. PMID- 11416437 TI - Electrocardiology Teacher Analysis and Review - Complete Interatrial Block. PMID- 11416438 TI - Images in Geriatric Cardiology - Left Sided Weakness and a History of Hypertension. PMID- 11416439 TI - Implantable Defibrillator Therapy for the Elderly. AB - The clinical application of ICD devices in the management of patients with ventricular tachyarrhythmias and cardiac arrest is rapidly expanding and is projected to exceed 50,000 implants annually within the next 5 years. Recent clinical trials have established superior clinical efficacy and safety with the ICD in these conditions as compared to currently available type 1 or type 3 antiarrhythmic drugs. Ventricular tachyarrhythmias are common in the elderly and most ICD centers have elderly patients. However, very limited clinical literature exists examining the specific considerations in implementing this therapy in the elderly patient and clinical expectations in this population. PMID- 11416440 TI - Drug Interactions in the Elderly With Coronary Artery Disease - Do These Interfere With Effective Therapy? AB - Older patients with cardiovascular disease receive multiple cardiac medications and often have multiple medical conditions for which they receive additional medical therapy. Over-the-counter medications are also frequently used in this older population. These factors coupled with altered drug elimination and potential alterations in drug responses in the elderly patient lead to increased opportunities for adverse drug interactions. In addition to the general geriatric dosing guidelines of starting medications at lower doses, increasing doses slower than in younger patients, and using as few drugs as possible, a framework for understanding drug-drug interactions is presented. Pharmacokinetic drug interactions alter drug concentrations and can either increase or decrease drug concentrations. Pharmacodynamic drug interactions alter the effects of the drug and can produce either additive or antagonistic effects. Examples of each interaction type are presented and currently available data on interactions of cardiovascular drugs with other drugs commonly administered in the elderly are briefly reviewed. PMID- 11416441 TI - Fibrinolysis for Acute Myocardial Infarction in the Elderly Patient: Is it Effective or Too Dangerous? AB - The incidence of myocardial infarction, and the subsequent mortality and morbidity increases markedly with age. Randomized placebo-contolled trials of fibrinolytic therapy have lacked adequate statistical power due to the relatively small numbers of older patients enrolled, but nevertheless have strongly suggested a mortality benefit in the older age group. Despite this, fibrinolytic therapy remains under-utilized in older patients, largely due to a perception that the risk, particularly of intracranial hemorrhage, outweighs any potential benefit of treatment. While the risk of intracranial hemorrhage following fibrinolysis does increase with age, the net clinical benefit, or the combined endpoints of death or disabling stroke, is still lower with treatment than without. Therapy with t-PA confers the greatest net clinical benefit in all but the very elderly. Age alone should not be a contraindication to fibrinolytic therapy for acute myocardial infarction. PMID- 11416442 TI - The Effect of Hypertension Treatment on the Prevention of Coronary Artery Disease Events in the Elderly. AB - Both isolated systolic (greater than 140/ less than 90 mm Hg) and systolic/diastolic ( greater than 140/ greater than 90 mm Hg) hypertension have been identified as major risk factors for cardiovascular disease in the elderly. Effective specific antihypertension therapy is available if lifestyle intervention fails to reduce blood pressure to normal levels. Long term placebo controlled trials, primarily with diuretics and beta-blockers, have demonstrated not only a reduction in cerebrovascular events but in CHD events as well. A significant reduction in the occurrence of CHF and progression of less severe to severe hypertension has also been demonstrated in treated compared to placebo subjects. A recent study suggests that the use of one of the moderately long acting dihydropyridine CCBs nitrendipine (not available in the U.S.) may also reduce the occurrence of strokes in elderly subjects with ISH PMID- 11416443 TI - Clinical Presentation of Coronary Artery Disease in the Elderly: How Does it Differ From the Younger Population? AB - CHD is the major cause of morbidity and mortality in the elderly in the U.S. In this age group, the clinical presentation of CHD can be quite atypical. In general, the incidence of typical precordial chest pressure/pain denoting myocardial ischemia is less common whereas dyspnea as an anginal equivalent symptom is frequent. The diagnosis of ischemic cardiac pain is frequently confused by the many comorbid conditions present in the elderly. Even when classic ischemic precordial discomfort is present it tends to be less severe and less well defined. The elderly appear to have reduced pain perception and as a result silent myocardial ischemia is more common and carries a somewhat worse prognosis in the elderly than in younger age groups. Similarly, the presenting symptoms of acute myocardial infarction in the elderly can be nonspecific. The classic crushing substernal chest pain decreases with age whereas the symptom of dyspnea gradually increases. Neurologic symptoms, weakness, and worsening heart failure are common clinical presentations of an acute infarction in elderly patients. Silent (unrecognized) myocardial infarctions are common in the elderly and carry serious prognostic implications. PMID- 11416445 TI - Risk Stratification in the Elderly Patient With Known or Suspected Coronary Artery Disease: Is it Useful; Who Should Have it? PMID- 11416444 TI - Myths and Facts About Coronary Disease in the Elderly. PMID- 11416446 TI - Myocardial Ischemia in the Asymptomatic Older Patient. AB - Although clinically manifest CAD is present in about one-fourth of individuals older than 65 years, a like proportion of additional persons have asymptomatic coronary artery stenoses, manifest only by inducible myocardial ischemia. A major challenge to contemporary cardiology is to detect the subset of such asymptomatic subjects at highest risk for future coronary events, especially myocardial infarction and sudden death. In the BLSA, we have employed various strategies toward this goal. In apparently healthy BLSA men and women, conversion from a normal to an ischemic ST segment response to treadmill exercise testing on serial biennial visits did not improve the prediction of subsequent coronary events. However, the combination of ischemic ST segment depression and a segmental thallium-201 perfusion defect induced by treadmill exercise identified a small subset of asymptomatic older subjects, 48% of whom developed a coronary event over a 4.6 year mean follow up period. The pathophysiology, risk factors, and treatment of silent myocardial ischemia are also briefly reviewed. PMID- 11416447 TI - Drug Treatment After Acute Myocardial Infarction: Is Treatment the Same for the Elderly as in the Young Patient? AB - The elderly population in our society experiences higher morbidity and mortality from cardiovascular disease. Risk reduction strategies for this group are based largely on the results of clinical trials in younger patients. The best evidence to date suggests that guidelines based on studies in younger populations can be extended to elderly patients. Yet recent studies indicate many elderly patients do not receive the benefit of these therapies. More comprehensive application of preventive medical therapies after myocardial infarction could have significant benefits for the elderly. PMID- 11416448 TI - Age Associated Alterations in Structure and Function of the Cardiovascular System. AB - The process of aging leads to significant changes in the structure and function of the cardiovascular system. Some of these changes result directly in pathological effects, while others serve to exacerbate the effects of cardiovascular diseases. The changes in the walls of the peripheral arteries lead to increased systolic and pulse pressures. In the myocardium, there is an age associated loss of myocytes. The remaining cells have reduced inotropic capacity. The aging heart also shows increased stiffness in both animal and human studies. Cardiac function is shown to be normal at rest in the aging population, however, during exercise, the aging heart shows a significantly smaller reserve. The increased mortality rates in setting of myocardial infarction and/or congestive heart failure in the elderly may be related to reduced muscle mass and loss of cardiac myocytes. Conversely, the aging population may receive greater benefit from the protective effects of beta-adrenergic antagonists and angiotensin converting enzyme inhibitors. PMID- 11416449 TI - The Effects of Aging and Endurance Exercise on Cardiovascular Performance in Healthy Elderly: A Review of the Literature. AB - CV performance declines over time in healthy elderly subjects due to aging, as well as declining physical activity. Recent studies with older, often sedentary individuals, demonstrate that aerobic endurance exercise may improve several measures of CV function. Although further research is needed to characterize the CV effects of exercise, particularly in elderly women and the oldest old population, aerobic endurance exercise should be advocated in the healthy elderly. PMID- 11416450 TI - Brief Report - Elderly Patients With Acute Myocardial Infarction: Need for New Strategies? PMID- 11416452 TI - Ethical Issues in the Management of Geriatric Cardiac Patients - A patient is asking his cardiologist to deactivate an implantable cardioverter defibrillator device in order to allow him to die. PMID- 11416451 TI - Images in Geriatric Cardiology - Diagnostic Advantage of TEE in Case of Aortic Disease and Stroke. PMID- 11416453 TI - Electrocardiology Teacher Analysis and Review - Peri-Infarction Block. PMID- 11416454 TI - Images in Geriatric Cardiology - Peripheral Edema, Shortness of Breath, and a Known Heart Murmur. PMID- 11416455 TI - Patent Ductus Arteriosus Diagnosed in Old Age. AB - Patent ductus arteriosus is a relatively common congenital heart disorder occurring in 0.5 percent of births and accounting for up to 10% of all congenital cardiac anomalies. Due to its early closure during infancy, however, patent ductus arteriosus is rare in the adult. A 76 year old woman was previously reported to be the oldest patient with a new diagnosis of patent ductus arteriosus. We present a woman first diagnosed at age 86. PMID- 11416456 TI - Factors Associated With Hospital Utilization in the Elderly: From the Cardiovascular Health Study. AB - OBJECTIVE: Analyze clinical, accepted biochemical, physiologic, and socioeconomic risk factors and correlate them with hospital utilization in an elderly population. DESIGN: Prospective, observational study in a defined, randomly recruited population. PARTICIPANTS: 5201 Medicare participants enrolled in the Cardiovascular Health Study (CHS). METHODS: Medicare recipients were randomly assigned to participate in an observational study. Baseline data were compared to hospital admissions and days of hospitalization over four years. DATA ANALYSIS: Data were grouped by type of risk factor and analyzed by Tobit analysis and logistic regression. RESULTS: Baseline variables associated with hospital use (p is less than 0.0001) were history of CHF, stroke, angina, hypertension, ln (timed walk), ln (blocks walked/week), age, gender, and clinic site. Factors not entering the model (p is greater than 0.05) were income, education, smoking, diabetes, weight, dietary fat, marital status, depression, and measures of mental function. CONCLUSIONS: In the elderly, existing health status is the major determinant of hospitalization and overwhelms many classic "risk factors" for morbidity. PMID- 11416457 TI - Acute Pulmonary Embolism Presenting as Pulmonary Hemorrhage/Infarction Syndrome in the Elderly. AB - PURPOSE: The purpose of this investigation was to determine the characteristics of the history, physical examination, chest radiograph, electrocardiogram, and the ventilation/perfusion lung scan probability in elderly patients with pulmonary embolism who present with the syndrome of pulmonary infarction or hemorrhage. BACKGROUND: In considering a possible diagnosis of acute pulmonary embolism, it is helpful to focus on the characteristics of the patient in terms of the characteristics of the presenting syndrome. The syndrome of pulmonary hemorrhage or pulmonary infarction is the most common syndrome of acute pulmonary embolism. It is defined by the presence of pleuritic pain or hemoptysis. Whether there are features of the pulmonary hemorrhage/infarction syndrome that are specific to elderly patients, or would suggest or tend to exclude a diagnosis is unknown. Therefore, this investigation was undertaken. METHODS: Patients reported in this investigation participated in the national collaborative trial of the Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED). All had pulmonary embolism diagnosed by pulmonary angiography. Patients with associated cardiac pulmonary disease were excluded. Elderly patients (N equals 17) were defined as patients at or above 70 years of age. Comparisons were made with patients age 40-69 years (N equals 67) and with patients under 40 years of age (N equals 35). RESULTS: In elderly patients, malignancy was a frequent predisposing factor (29%) in addition to immobilization (76%). Dyspnea or tachypnea, in addition to pleuritic pain or hemoptysis, were present in 94% of elderly patients. Among elderly patients with the pulmonary hemorrhage/infarction syndrome, pleuritic pain was more frequent than hemoptysis (88% vs. 12%). The electrocardiogram was normal in 62% of elderly patients. If abnormal, the most frequent abnormalities were nonspecific ST segment or T-wave changes (38%). The chest radiograph showed atelectasis or a pulmonary parenchymal abnormality in 82% of elderly patients. The central pulmonary artery dilated in 29% of elderly patients. A normal chest radiograph was uncommon, occurring in only 6%. The ventilation/perfusion lung scan was useful and was interpreted as high probability for pulmonary embolism in 41% of elderly patients. Elderly patients had a higher pulmonary artery mean pressure (25 A+/-9 vs. 17 A+/-7 mm Hg) and lower partial pressure of oxygen in arterial blood (64 A+/-10 vs. 81 A+/-14 mm Hg) than patients less than 40 years, and elderly patients tended to have more mismatched segmental perfusion defects on the ventilation/perfusion lung than patients less than 40 years (4.8 A+/-5.5 vs. 2.1 A+/-2.9 mismatched perfusion defects). CONCLUSIONS: Although a few clinical features of the pulmonary hemorrhage/infarction syndrome of pulmonary embolism in elderly patients differ from patients less than 40 years of age, in general, the clinical characteristics are comparable to younger patients. PMID- 11416458 TI - Electrocardiology Teacher Analysis and Review - Hypokalemia. PMID- 11416459 TI - Ethical Issues in the Management of Geriatric Cardiac Patients - A patient with a living will develops drug-induced Torsades-de-Pointes. PMID- 11416460 TI - Spectral Turbulence Analysis: A Valuable Method for the Prediction of Cardiac Events in Elderly Patients With Intraventricular Conduction Abnormalities. AB - BACKGROUND: Intraventricular block and its associated heart disease are common in the elderly. The aim of this study was to evaluate the ability of spectral turbulence analysis of signal-averaged electrocardiograms to predict the cardiac events of sudden death, sustained ventricular tachycardia, ventricular fibrillation, and syncope in elderly patients with wide QRS as seen on a standard 12 lead surface electrocardiogram. METHODS: We studied 51 consecutive elderly patients (age at or above 70 years), for whom at least 12 months of follow up was available following the initial signal-averaged electrocardiogram. The patients included 40 males, and 11 females, of whom 47 had coronary artery heart disease and 25 had a history of prior myocardial infarction, and 4 with dilated cardiomyopathy. The subjects were divided into 3 groups. Group I comprised 22 patients without any events; Group II included 16 patients with events-sudden death(N equals 6), ventricular tachycardia (N equals 5), ventircular fibrillation (N equals 1), and syncope (N equals 4); Group III consisted of 13 patients with nonsustained VT. The bipolar three-lead orthogonal signal-averaged electrocardiogram from each patient was analyzed for spectral turbulence. The optimal criteria for predicting cardiac events were determined by computer algorithm. RESULTS: Spectral turbulence predicted future cardiac events with a sensitivity of 88%, a specificity of 67%, a positive predictive accuracy of 67%, a negative predictive accuracy of 88%, and a total predictive accuracy of 74%. CONCLUSIONS: Spectral turbulence analysis of the signal-averaged electrocardiogram was found to be a highly sensitive predictor of serious cardiac events in elderly patients with a wide QRS complex. PMID- 11416461 TI - Should Orthostatic Changes in Blood Pressure be Measured After One or Three Minutes in Elderly Subjects? AB - Orthostatic hypotension is common in the elderly. Standard clinical practice is to measure changes in SBP after 1 minute or 3 minutes of upright posture. In an elderly patient, the lowest blood pressure may occur early after the assumption of upright posture and recover. To address this issue, we evaluated beat to beat changes in SBP and heart rate during the first 3 minutes of upright posture on a tilt table in a cohort of community-dwelling elderly subjects. Almost all of the subjects have significant declines in SBP with upright posture. While the majority of the transient hypotensive episodes are not detected by the standard clinical approach, measurement of SBP after 1 minute of upright posture was twice as sensitive as a single measurement at 3 minutes. Use of continuous beat to beat measurement of SBP identified an even greater number of elderly subjects who have transient orthostatic hypotension. PMID- 11416462 TI - Understanding and Managing Isolated Systolic Hypertension in the Elderly. AB - In summary, ISHT in the elderly is a result of the interaction of increased systemic vascular resistance and of progressive arterial stiffening. The early return of reflected pulse wave may add to peak SBP in the ascending aorta and central arteries, thereby increasing LV systolic stress (afterload). Antihypertensive therapy has been shown to be effective in the prevention of strokes associated with ISHT. The reduction in the incidence of fatal and non fatal MI by active treatment may be limited by excessive lowering of DBP, resulting in inadequate myocardial perfusion in some patients (J-curve). The ideal drug for the treatment of ISHT should reduce systemic vascular resistance, increase large artery compliance, and delay the return of reflected wave in central arteries. Diuretics effectively lower SBP in the elderly with ISHT. Long acting nitrates lower SBP selectively and have a favorable effect on LV systolic stress (afterload). Baroreceptor dysfunction due to increased arterial stiffness increases the risk of orthostatic and postprandial hypotension in the elderly during antihypertensive therapy. PMID- 11416463 TI - Treatment of High Blood Pressure in the Elderly: A Position Paper from the Society of Geriatric Cardiology. PMID- 11416464 TI - What Older Adults Should Know About High Cholesterol: A Position Paper from the Society of Geriatric Cardiology. PMID- 11416465 TI - Physical Activity and Exercise Training in the Elderly: A Position Paper from the Society of Geriatric Cardiology. PMID- 11416466 TI - Ethical Issues in the Management of Geriatric Cardiac Patients - A family member with power of attorney for an 87 year old patient is requesting removal of the patient's pacemaker. PMID- 11416467 TI - Electrocardiology Teacher Analysis and Review - Atrial Tachycardia vs. Junctional Tachycardia and Partly Concealed P' Waves. PMID- 11416468 TI - Integration of Geriatrics into Cardiology Fellowship Training Programs: A Joint Position Paper from the American Geriatrics Society and the Society of Geriatric Cardiology. PMID- 11416469 TI - Atrial Fibrillation in the Elderly. PMID- 11416470 TI - Atrial Fibrillation in the Elderly: Who is at Risk? AB - Atrial fibrillation is recognized by all clinicians as an important arrhythmia. Its importance relates to the increased risk for cardioembolic stroke. The elderly are the most commonly affected. They have the highest incidence of atrial fibrillation and are the patients most vulnerable to the risk of stroke. It has been stated that the incidence of atrial fibrillation is only 0.2 per 1000 in patients aged 30-39. This figure increases to 39.0 per 1000 in patients 80-89 years old. Among patients seeking medical attention the prevalence can be even greater (Fig. 1). Atrial fibrillation's contribution to the risk of stroke increases with age. In patients 50-59 years of age it is estimated that 6.7% of strokes are associated with atrial fibrillation. In patients aged 80-89 this figure increases to 36.2%.1 This review will explore common etiologies leading to atrial fibrillation in the elderly. Important reversible causes will be emphasized. Some of the reasons this common arrhythmia is an especially important risk factor for cardioembolic stroke in this group will be illustrated. PMID- 11416471 TI - The Role of Echocardiography in the Management of Atrial Fibrillation. AB - From the time of its introduction as a clinical tool in the early 1970s, efforts have been made to use echocardiography to characterize the atrium and its function as these relate to attempted cardioversion and embolic risk. M-mode and 2-D echo studies suggest left atrial size is not a definitive determinant of the potential for successful cardioversion. Doppler studies show that atrial function can take weeks to recover postcardioversion, accounting in part for the delayed embolic risk. TEE is particularly effective at detecting atrial thrombi and shows that slow velocities in the atrium and appendage are an important risk factor for embolism and clot. All are associated with the finding of spontaneous echo contrast and atrial stunning postcardioversion. The ultimate role of TEE in management is not yet fully defined. The insights these and other studies provide to our understanding of atrial physiology and the appropriate management of atrial fibrillation are reviewed. PMID- 11416472 TI - Long Term Management of Atrial Fibrillation: Maintenance of Sinus Rhythm vs. Rate Control and Anticoagulation. AB - Two strategies for treatment of atrial fibrillation in the elderly include rate control with anticoagulation, or cardioversion and maintenance of sinus rhythm. Rate controlling agents include digoxin, calcium blockers, and b-blockers, most effective during exercise, or nonpharmacologic control with AV nodal ablation and pacemaker implantation. Agents to maintain sinus rhythm in the elderly include class Ia and III antiarrhythmics, with 1 year efficacies of approximately 50%. Whether the overall benefits of pharmacologic maintenance of sinus rhythm outweigh the risks of proarrhythmia and other adverse effects is under debate, but will be better delineated by the ongoing AFFIRM trial and VA cooperative trial, both randomizing patients to one of these two strategies. PMID- 11416473 TI - Prevention of Stroke. AB - Patients with atrial fibrillation are at variable risks for developing a stroke. The risk is significantly increased among the elderly. Other risks factors include hypertension, diabetes, poor ventricular function, and a prior history of TIA or stroke. Among the elderly, it is generally recommended that those patients who are eligible for anticoagulation would benefit substantially from its use provided the INR is maintained between 2.0-3.5 and blood pressure is well controlled. In those patients that cannot be safely anticoagulated, aspirin at a dose of 325 mg is a viable alternative but is likely to offer substantially less protection than warfarin. PMID- 11416474 TI - Ethical Issues in the Management of Geriatric Cardiac Patients - An elderly patient declines cardiac surgery under optimal circumstances yet undergoes cardiac surgery when in extremis. PMID- 11416475 TI - Electrocardiology Teacher Analysis and Review - Accelerated A-V Junctional Rhythm. PMID- 11416476 TI - Diagnosis and Treatment of Systolic Heart Failure in the Elderly. AB - Appropriate therapy of systolic LV dysfunction in the elderly depends on the stage of the disease process. Evidence of systolic dysfunction is best detected by clinical assessment and echocardiographic exam. While one seeks to define and treat the factors that initiated the process, one also attempts to reduce symptoms and prolong life. Therefore, with initial damage (e.g., following a large myocardial infarction), ACE inhibition or angiotensin II receptor blockade is indicated. A -Adrenergic receptor blockade is also indicated at this stage because of mortality reduction. As failure progresses to more symptomatic phases, A -blockers also appear along with ACE inhibition. Once symptoms increase and peripheral edema and central congestion occur, loop diuretics become useful to maintain dry weight. In order to prevent further myocardial cell loss, other measures, such as cessation of smoking and treatment of hypertension, are essential. Digitalis glycosides, especially in modest doses, are indicated when NYHA Class II and III symptoms occur. Surgical approaches, such as coronary revascularization, may be performed to improve LV function in ischemic disease. Therapy of heart failure seeks the reversal or attenuation of the process that initiated the syndrome, while treating the patient to relieve symptoms and prolong life. Even in very late stages of the disease, relief of symptoms can now be accomplished with modest gains in life expectancy. PMID- 11416477 TI - Nonpharmacologic Therapy of Atrial Fibrillation and Atrial Flutter. AB - The medical therapy of atrial fibrillation and atrial flutter is unsatisfactory for many patients since nonpharmacologic therapy is often necessary. Curative catheter ablation is the treatment of choice for atrial flutter. Primary success rates of 95% are routinely achieved in centers of excellence with relapse rates for atrial flutter of 5%-10%. Nonpharmacologic therapies in atrial fibrillation are more often palliative than curative. Catheter ablation of the AV junction with implantation of a permanent pacemaker is the primary nonpharmacologic therapy for medically refractory atrial fibrillation. The quality of life is improved by AV junction ablation, although survival is not. Curative therapy for atrial fibrillation is available with the surgical maze operation but that is primarily offered in conjunction with concomitant cardiac surgery. Recently a catheter ablative procedure to mimic the maze operation has had preliminary evaluation but remains investigational. A focal ablation procedure for paroxysmal atrial fibrillation has also entered into clinical trials. PMID- 11416478 TI - Treatment of Older Patients With CHF Associated With Normal LVEF. AB - CHF with a normal LVEF should be treated with a low salt diet, cautious use of loop diuretics, and A -blockers. If CHF persists despite the use of diuretics and A -blockers, ACE inhibitors should be administered. If the patient cannot tolerate ACE inhibitors because of cough, rash, or altered taste sensation, angiotensin II type 1 receptor antagonists should be given. If CHF persists despite treatment with diuretics, A -blockers, and ACE inhibitors, isosorbide dinitrate plus hydralazine should be given. If the patient cannot tolerate A blockers, ACE inhibitors, and angiotensin II type 1 receptor antagonists, isosorbide dinitrate plus hydralazine should be administered. If CHF persists despite the use of diuretics and the patient is unable to tolerate A -blockers, ACE inhibitors, and isosorbide dinitrate plus hydralazine, calcium channel blockers are preferred. Digoxin should be avoided if sinus rhythm is present. PMID- 11416479 TI - Cardiovascular Aging: Perspectives From Humans to Rodents. AB - In order to define and target the specific characteristics of cardiovascular aging that render aging as a risk factor for diseases (such as atherosclerosis, hypertension, stroke, and heart failure) that reach epidemic proportions among older individuals, it is essential to develop quantitative information on age associated alterations in cardiovascular structure and function in health. A sustained effort, over the past two decades, has been applied to characterize the multiple effects of aging on health in cardiovascular structure and function in a single study population (BLSA). In these studies, community-dwelling, volunteer participants are rigorously screened to detect both clinical and occult cardiovascular disease and characterized with respect to lifestyle (e.g., exercise habits), in an attempt to deconvolute interactions among lifestyle, cardiovascular disease, and the aging process in health. Some specific changes in resting cardiovascular structure and function and cardiovascular reserve capacity that occur with advancing age in these healthy humans have been identified and are presented here. These observations in humans are extended by relevant experiments from animal models to provide possible mechanistic insight. PMID- 11416480 TI - Cardiopulmonary Resuscitation in the Elderly: Defining the Limits of Appropriateness. AB - The prevailing policy of treating people to their death and to resuscitate victims of cardiopulmonary arrest regardless of their health status is both inhumane and irrational. In this review we will explore how to change this wasteful high technology-driven, death-denying culture into one that accepts the eventuality of dying. PMID- 11416481 TI - Antiarrhythmic Drug Therapy of Ventricular Tachycardia in the Elderly: Lessons From Clinical Trials. AB - Life expectancy is increasing even though life span is fixed. There will be almost 35 million Americans over 65 years of age in the year 2030. CAD will account for almost 80% of the deaths in the population with almost 50% of such deaths classified as sudden. Ventricular arrhythmias increase with age and have been shown to be independent markers for premature cardiac death. Therefore, many trials involving the use of antiarrhythmics have been performed, hoping to show an improvement in overall survival. Unfortunately, some of the classical antiarrhythmic agents despite markedly suppressing the arrhythmias, have been shown to be harmful or of no benefit. The use of b-blockers, amiodarone, aspirin, and ACE inhibitors show some potential for benefit in this high risk patient population. The implantable devices are superior to antiarrhythmics in survivors of cardiac arrest. PMID- 11416482 TI - Age and Quality of Life. PMID- 11416483 TI - Determining Quality of Life in Older and Younger Women With Congestive Heart Failure and Myocardial Infarction. AB - The purpose of this study was to compare quality of life (QOL) between older and younger women diagnosed with congestive heart failure (CHF) or myocardial infarction (MI) and to ascertain if QOL varied with age. Using the Medical Outcomes Survey (MOS-SF-36) we examined 131 women: 69 diagnosed with CHF and 62 with MI. Older women (age equals 65 years) with CHF had a higher mean for general health, whereas younger women (age equals 64 years) had higher mean scores for role physical, vitality, role emotion, and social functioning. Older women with MI had higher mean scores on role emotion, mental health, and social functioning. The younger MI women had higher mean scores on physical functioning, and role physical. Aspects of QOL may vary in subsets of women depending on age and cardiac diagnosis. This suggests the need for age appropriate intervention in women with MI and CHF. PMID- 11416484 TI - A Five Year Study of Comparison of Characteristics and In-hospital Complications of Acute Myocardial Infarction Between Elderly and Younger Patients. AB - Elderly patients suffering from acute myocardial infarction (AMI) can present differently and their outcome may be different from those of younger patients. A prospective study looking into the characteristics and in-hospital complications of AMI in the elderly patients, comparing to those of the younger patients, was conducted. All AMI patients admitted into our hospital from April 1991 to March 1996 were enrolled. Those patients greater than 70 years were classified as elderly patients. During the 5 year period, there were 241 elderly AMI patients (mean age of 77 years) and 357 younger patients (mean age of 60 years). In the elderly group, 48.0% were female as compared to 24.0% in the younger group (p is less than 0.0001). For the in-hospital complications, 29.5% of elderly patients had cardiogenic shock as compared to 14.6% in younger patients (p is less than 0.0001). There were also significantly more elderly patients suffering from atrial arrhythmias (11% vs. 6% in younger patients, p is less than 0.05). Both groups of patients had similar incidence of ventricular arrhythmias and various types of heart block. Health care professionals taking care of these patients should be aware of these differences and well prepared to tackle these complications. PMID- 11416485 TI - Reducing Mortality Among Elderly Patients Undergoing Urgent Surgical Revascularization for Acute Myocardial Infarction. AB - INTRODUCTION: Elderly patients undergoing coronary bypass surgery after acute myocardial infarction represent a distinct, high risk subgroup. We sought to identify independent risk factors for mortality in a series of patients operated on in our hospital. METHODS: The case records of 499 consecutive patients greater than 70 years were identified, and 94 of these patients underwent urgent/emergent coronary bypass surgery within 7 days of acute myocardial infarction. Patients received either cold (4A degrees C) or tepid (32A degrees C) cardioplegia for myocardial protection. RESULTS: Mortality for the tepid cardioplegia group was 0/33, and the mortality for the cold cardioplegia group was 12/61 (0% vs. 20%, p equals 0.003). Multivariate analysis demonstrated left ventricular dysfunction and cold blood cardioplegia to be independent predictors of mortality. CONCLUSION: The type of myocardial protection technique is a significant predictor of mortality, and tepid cardioplegia may provide significant advantages to this high risk patient population. PMID- 11416486 TI - Direct Angioplasty for Acute Myocardial Infarction in Elderly Patients Using Transradial Approach. AB - BACKGROUND: In acute myocardial infarction (AMI), immediate outcome seems to be better after direct coronary angioplasty (DCA) than thrombolysis. Right transradial approach (RTRA) has been currently advocated as an alternative catheterization method for coronary angioplasty, but it is an exceptional technique in the treatment of AMI. PURPOSE: The goal of this prospective study was to evaluate feasibility and safety of DCA using the RTRA in elderly patients. METHODS: All patients, 70 years or older with AMI (less than 6 hours after the onset of symptoms), were included in this study. Exclusion criteria were ischemic Allen Test, and previous thrombolytic treatment. Left heart catheterization, with selective coronary angiograms were realized in all of them, using RTRA. RESULTS: Forty six patients, (mean age 76A+/-5, range 70-85), (31 men, 15 women), were included. Coronary catheterization using RTRA was not possible in 4 patients, (9.1%), and DCA was not necessary in 2 other patients with two distal lesions (4.5%). In the 40 remaining patients, transradial coronary angioplasty of the infarct related vessel could be successfully achieved in 40 patients, (100%) with balloon alone in 21 patients, (52.5%), or stents in 19 patients, (47.5%). Two patients with cardiogenic shock died (in hospital mortality: 4.5%). In 2 patients, recurrent angina needed new angioplasty using the same RTRA (4.5%), and in 2 other patients, asymptomatic postprocedural right radial artery occlusion was observed. Before the 24th hr, 30/40 patients (75%) could stand up with an early discharge, (7A+/-4 days). CONCLUSION: DCA using RTRA for AMI in elderly patients can be easily realized, with a high rate of success, without any more major local complications than using the usual femoral route, allowing earlier getting up and discharge. PMID- 11416487 TI - Ethical Issues in the Management of Geriatric Cardiac Patients - The wife of a hopelessly terminal patient asks physicians to hasten the moment of death. PMID- 11416488 TI - Images in Geriatric Cardiology - Giant Left Ventricular Pseudoaneurysm. PMID- 11416489 TI - Electrocardiology Teacher Analysis and Review - Atrial Flutter vs. (Paroxysmal) Atrial Tachycardia With Block. PMID- 11416492 TI - Resistance Exercise in the Elderly Person with Cardiovascular Disease. AB - Elderly cardiovascular disease patients are not aggressively referred to cardiac rehabilitation programs due to concerns regarding orthopedic fragility, hemodynamic risk, and life expectancy. However, there is a growing consensus that older cardiovascular patients enrolled in rehabilitation programs will derive the same magnitude of improvement in functional capacity as their younger counterparts. Further, resistance exercise training may be necessary, as a part of a comprehensive cardiac rehabilitation program, to elicit optimal benefits in elderly cardiovascular patients. In this article, we briefly review orthopedic and hemodynamic risks associated with resistance training, the efficacy of resistance training, and the recommended guidelines for resistance exercise prescription. (c)1999 by CVRR, Inc. PMID- 11416493 TI - Aggressive Risk Factor Management in the Elderly: Are You Ever Too Old? AB - Smoking, hypertension, and hyperlipidemia are the three most important modifiable risk factors contributing to the development of cardiovascular disease in older adults. Although the magnitude of risk associated with smoking and hyperlipidemia declines with age, the absolute number of cases attributable to these risk factors increases due to the increasing prevalence of cardiovascular disease. Smoking increases the risk of both coronary events and stroke in the elderly, and there is evidence that smoking cessation is associated with a rapid reduction in risk. Therefore, an aggressive effort to promote smoking cessation is strongly recommended in patients of all ages. Systolic and diastolic hypertension are powerful risk factors for cardiovascular disease in the elderly. Moreover, multiple clinical trials have demonstrated that blood pressure reduction reduces the risk of stroke, coronary events, heart failure, and cardiovascular death in individuals at least up to the age of 90. Accordingly, treatment of both systolic and diastolic hypertension are strongly recommended regardless of patient age. The importance of total serum cholesterol as a coronary risk factor declines with age, but the ratio of low density lipoprotein cholesterol (LDL-C) to high density lipoprotein cholesterol (HDL-C) remains an independent predictor of coronary events in older men and women. In addition, clinical trials have shown that cholesterol reduction is associated with improved clinical outcomes in individuals at or above 75 years of age. At the present time, the value of treating hyperlipidemia in patients greater than 80 years of age is unknown, and therapy in this age group must be individualized. (c)1999 by CVRR, Inc. PMID- 11416494 TI - Chronic Heart Failure and Cardiac Rehabilitation for the Elderly: Is It Beneficial? AB - Heart failure is a common health problem among persons at or above 65 years of age. To date, no data exist describing to what extent, if any, cardiac rehabilitation or exercise training might be useful in this population. Fourteen randomized trials involving mostly heart failure patients less than 65 years of age showed that exercise training improves exercise tolerance and reverses, in part, abnormalities in both autonomic function and skeletal muscle histology and biochemistry. Based on these trials, the use of exercise training in older persons with New York Heart Association (NYHA) Class II or III heart failure appears to be a safe and effective adjunctive therapy. Exercise trials involving patients with heart failure at or above 65 years of age are necessary. (c)1999 by CVRR, Inc. PMID- 11416495 TI - Brief Report - SAFE PACE 2 Syncope And Falls in the Elderly Pacing And Carotid Sinus Evaluation: A Randomized Control Trial of Cardiac Pacing in Older Patients With Falls and Carotid Sinus Hypersensitivity. PMID- 11416496 TI - Prescription Drugs for Elderly Patients and Medicare Reform. PMID- 11416497 TI - Cardiostim Congress - Implantable Cardioverter Defibrillators and Pacemakers in the Elderly. PMID- 11416498 TI - Syncope Evaluation in the Elderly. AB - Syncope is a common and often recurrent clinical problem in elderly patients. Frequently, the precise etiology of recurrent syncope in the elderly remains unclear despite repeated diagnostic testing. This leads to frustration, not only for patients and family members, but also for the treating physician. In many cases, the cause of syncope in the elderly is multifactorial. Recurrent episodes necessitate repeated hospitalization and lead to considerable morbidity and increased cost. Despite these diagnostic problems, however, most episodes of syncope in elderly patients can be classified as cardiogenic or noncardiogenic on the basis of a thorough history and careful physical examination. Cardiogenic causes can be further divided into reflex, orthostatic, and primary cardiac disorders, allowing for more focused diagnostic testing. In this review, the etiology, pathophysiology, and distinguishing clinical features of syncope in the elderly, as well as current therapeutic strategies in patients with recurrent syncope, are discussed. (c)1999 by CVRR, Inc. PMID- 11416499 TI - Pacing and Implantable Cardioverter Defibrillator Options for the Elderly in Australia and New Zealand. AB - This review seeks to determine the extent to which age impacts the access to pacemaker and implantable cardioverter defibrillator (ICD) technology in Australia and New Zealand. The limited available data suggest a number of consistent trends: 1) implant rates for both pacemakers and ICDs are increasing in both Australia and New Zealand; 2) the elderly are increasingly well catered for in terms of pacemaker implantation, especially in Australia (54% of pacemaker recipients aged at or above 80 in 1997); 3) there is a progressive trend to more dual chamber pacing in the elderly in both countries; 4) age appears to be a barrier to the access to ICDs, especially in New Zealand, which is likely related to the high cost of these devices; and 5) limited data suggest that Australia physicians consider age less of a barrier to the access to high technology implantable devices, compared with their New Zealand counterparts, likely reflecting their greater prosperity and financial commitment to health care. Finally, this review clearly documents the need for improved prospective data collection in both countries. (c)1999 by CVRR, Inc. PMID- 11416500 TI - Coil Occlusion of Elderly Ductus Arteriosus. AB - Surgical management of patent ductus arteriosus (PDA) in the elderly population is complicated by a fragile aortic wall due to atheromatous lesions, the presence of friable tissue at the surgical site, and calcification of the ductus. A viable option for the elderly patient is transcatheter closure of the PDA using a synthetic plug. Transcatheter coil occlusion has been shown to be an effective nonsurgical technique for closure of the persistently PDA in a younger population. We report a case of a 75 year old woman who had complete occlusion of her PDA with a single Gianturco coil after recurrence of her ductus 3 years post surgical ligation. The successful transcatheter treatment is presented with emphasis on the simple and safe technique utilized. (c)1999 by CVRR, Inc. PMID- 11416501 TI - The Great Debate: Aggressive vs. Conservative Management of Coronary Artery Disease in Elderly Patients. AB - You wake up in a cold sweat. You dreamt that the country in which you practice rations aggressive care and insists on an evidence based justification for each cardiac procedure. You are allowed only one coronary angiogram each day, one balloon catheter, one stent. You have 2 patients, each admitted that day with acute myocardial infarction (MI). They are identical, except for their age; one is 42 years old, the other is 84. For the last several hours, you have tossed and turned, embroiled in a great debate. Which one should be treated aggressively? (c)1999 by CVRR, Inc. PMID- 11416502 TI - Images in Geriatric Cardiology: Ruptured Chordae Tendineae Complicating Mitral Valve Prolapse. PMID- 11416503 TI - Electrocardiology Teacher Analysis and Review: Atrial Flutter vs. (Paroxysmal) Atrial Tachycardia With Block. PMID- 11416505 TI - Prescription Drugs For The Elderly: How Physicians Can Help. PMID- 11416504 TI - Ethical Issues in the Management of Geriatric Cardiac Patients - A patient with clear, advance instructions to forego placement on a ventilator dies after several weeks of intensive care. PMID- 11416506 TI - Brief Report - International Initiatives to Improve the Quality of Health in the Elderly. PMID- 11416507 TI - Perceptions of Women Living With Coronary Heart Disease: An Overview of Study Findings. AB - OBJECTIVE: To describe how 41 perimenopausal and postmenopausal women with coronary heart disease (CHD) perceive their illness experience. METHODS: Five focus groups were conducted at two community hospitals in New Jersey. Predetermined questions explored psychosocial and gender sensitive issues. Data collection and analysis was based on the grounded theory. RESULTS: Participants often failed to recognize the significance of risk factors and symptoms, and even denied the presence of disease following diagnosis and treatment. Consequences of illness centered on women's efforts to maintain control over their health and their lives. A gap existed between the desire for knowledge and the ability to access and assimilate information in key areas regarding CHD and personal health behaviors. CONCLUSIONS: Although not generalizable to the population at large, the insights provided by these women have several implications for future research and contribute valuable information related to the needs of this population. (c)1999 by CVRR, Inc. PMID- 11416508 TI - Cardiopulmonary and Cardiovascular Physiology in Elderly Competitive Endurance Athletes. AB - Thirteen elderly male high intensity endurance athletes and 12 healthy age matched nonathletes were evaluated to determine differences in their cardiovascular physiology. Contrary to several pervious reports, this study did not demonstrate superiority of left ventricular (LV) systolic or diastolic function in athletes compared with controls, either at rest or immediately following exercise. Fasting serum lipids failed to demonstrate a favorable cholesterol profile in the athletes compared to inactive controls. Our results point to superior oxygen utilization (mean peak Vo2 44 vs. 27 mL/kg/min; athletes vs. controls) and the capacity to exercise both aerobically (mean duration 24 vs. 12 min; athletes vs. controls) and anaerobically (mean duration 10 vs. 4 min; athletes vs. controls) as distinguishing parameters between the two cohorts. Our study suggests that in senior athletes, enhanced LV diastolic function may not be as great a contributing factor to athletic performance as previously reported. (c)1999 by CVRR, Inc. PMID- 11416510 TI - Observations on the Heart - Senile Cardiac Calcification Syndrome. PMID- 11416509 TI - Early and Intermediate Outcomes After Rotational Atherectomy in Octogenarian Patients. AB - OBJECTIVE: To provide information about the use of rotational atherectomy in octogenarians. MATERIAL AND METHODS: From 1993-1996, 26 octogenarians underwent coronary intervention with rotablator at Emory University Hospitals. The total number of lesions were 28 (57% of them in the left artery descending). Adjunctive balloon angioplasty was utilized in 26 lesions. The lesions were eccentric in 82%, calcified in 75%, with a mean length of 10.2A+/-8.5 mm. RESULTS: The angiographic success rate was 96.2%. The only in-hospital complications was the death of 1 patient (3.8%) after an abrupt closure, and a dissection of 1 lesion of another patient, with a favorable subsequent outcome. The mean follow up was 1.3A+/-0.4 years. The Kaplan-Meier survival rate at 1 year was 96.2% and at 2 years 78.3%. The freedom from death, myocardial infarction (MI), and coronary artery bypass graft (CABG) was 88.5% at 1 year and 59.7 % at 1.5 and 2 years. The freedom from death, MI, CABG, and percutaneous transluminal coronary angioplasty (PTCA) was 69.2% at one year and 36.3% at 1.5 and 2 years. CONCLUSIONS: In octogenarians with complex lesions, rotablator was performed in conjunction with balloon angioplasty, and showed a high rate of angiographic success, a relatively low complication rate, and a favorable 2 years survival rate. Nevertheless, the event free rates did not show the same favorable evolution after 1 year. If feasible, it would be useful to conduct randomized trials in the elderly to compare interventional procedures to medical treatment or surgery. (c)1999 by CVRR, Inc. PMID- 11416511 TI - Images in Geriatric Cardiology - Type II Aortic Dissection. PMID- 11416512 TI - Electrocardiology Teacher Analysis and Review - Global T Wave Inversion. PMID- 11416513 TI - Ethical Issues in the Management of Geriatric Cardiac Patients - A "slow code" is suggested in response to a critical situation involving a patient with multiple medical problems. PMID- 11416514 TI - Cardiac Imaging in Elderly Patients. PMID- 11416516 TI - Cardiac Disease in the Elderly: Radiographic Features. AB - The plain film appearances of cardiac disease in the geriatric age group are often different in younger patients because of the increased incidence of age related heart diseases, the physiological alterations that affect the range of normal values in the elderly, and the complicating effects of age related noncardiac diseases. Although the clinical significance of conditions, such as coronary artery calcification, calcification of the mitral annulus, and senile calcification of the aortic valve, is not completely understood at the present, they are indicative of significant disease in an appreciable number of patients. Furthermore, the roentgen appearance of the normal heart and lungs, as well as common cardiac conditions such as congestive failure, can be different in the elderly. If these changes are not appreciated, interpretation of the ubiquitous chest film, one of the most common and available techniques for the detection of heart disease, can result in misdiagnosis as well as a missed diagnosis. (c)1999 by CVRR, Inc. PMID- 11416517 TI - Measurement of Coronary Artery Calcium in Elderly Patients. AB - Quantification of coronary artery calcium by electron beam and spiral computed tomography (CT) is being increasingly utilized as a technique to assess the presence and extent of coronary artery disease (CAD) as well as the prognosis for clinical events in both symptomatic and asymptomatic individuals. Much of the demand for this procedure is generated by patients exposed to marketing efforts, and patients often ask physicians about the value of this test. Since elderly patients have a high prevalence of coronary artery calcium as well as a higher incidence of clinical CAD, the interpretation and clinical utility of this test differs from that for younger individuals. This paper attempts to review currently available information concerning coronary artery calcium quantification and its potential uses in elderly patients. (c)1999 by Quantification of coronary artery calcium by electron beam and spiral computed tomography (CT) is being increasingly utilized as a technique to assess the presence and extent of coronary artery disease (CAD) as well as the prognosis for clinical events in both symptomatic and asymptomatic individuals. Much of the demand for this procedure is generated by patients exposed to marketing efforts, and patients often ask physicians about the value of this test. Since elderly patients have a high prevalence of coronary artery calcium as well as a higher incidence of clinical CAD, the interpretation and clinical utility of this test differs from that for younger individuals. This paper attempts to review currently available information concerning coronary artery calcium quantification and its potential uses in elderly patients. (c)1999 by CVRR, Inc. PMID- 11416518 TI - Magnetic Resonance of Coronary Artery Disease in the Elderly. AB - Coronary artery disease is one of the leading causes of morbidity and mortality in the elderly population. Currently, the work up of coronary artery disease requires an integrated approach utilizing several diagnostic imaging modalities including coronary x-ray angiography, echocardiography, and radionuclide scintigraphy. Magnetic resonance imaging (MRI) for cardiac imaging offers great potential in the work up of ischemic heart disease. MRI allows the evaluation of the coronary arteries, quantitation of coronary flow and flow reserve, and determination of viability of ischemic myocardium. This paper reviews established and future applications of cardiac MRI with special emphasis on ischemic heart disease. (c)1999 by CVRR, Inc. PMID- 11416519 TI - Noninvasive Imaging of the Thoracic Aorta. AB - Thoracic aortic disease is a common problem affecting the geriatric population of the U.S. Thoracic aortic aneurysms and aortic dissection and its variants are the most common diseases of the thoracic aorta, often leading to death if not treated immediately. Contrast aortography is no longer considered to be the diagnostic gold standard for evaluating the thoracic aorta. This is a review of those abnormalities most commonly seen in the geriatric population: thoracic aortic aneurysms, aortic dissections, intramural hematomas, and penetrating ulcers. The roles of multiplanar transesophageal echocardiography (TEE), spiral computed tomography (CT), and magnetic resonance imaging (MRI) are discussed in the evaluation of each of these disease entities. Although each has been shown to have high sensitivities and specificities for the diagnosis of aneurysms and dissections and its variants, TEE and MRI are thought to be superior to spiral CT because of their ability to evaluate the aortic valve apparatus. However, spiral CT is perhaps the easiest and most cost effective modality to screen patients with known or suspected aortic dissection. (c)1999 by CVRR, Inc. PMID- 11416521 TI - Electrocardiology Teacher Analysis and Review - Wolff-Parkinson-White (WPW) ECG With Deceptive Features. PMID- 11416520 TI - Helical CT Angiography of the Pulmonary Arteries: Changing the Diagnostic Approach to Patients With Suspected Pulmonary Embolism. AB - Helical (spiral) computed tomography (CT) enables angiogram like images of the pulmonary circulation to be acquired in a single breath hold and has revolutionized the diagnostic work up in patients with suspected pulmonary embolism (PE). The sensitivity and specificity of CT exceed that of scintigraphy, and the accuracy is similar to that of pulmonary arteriography through the segmental arterial level. Unlike other diagnostic tests, CT can also depict clinically significant nonembolic intrathoracic disease, common in elderly patients. This review discusses the technique of helical CT pulmonary arteriography, criteria for interpretation, and its clinical application with particular emphasis on problems common in the geriatric population. CT has numerous advantages over scintigraphy, and should be used as the initial test in patients with suspected PE, particularly those with associated cardiac or pulmonary disease. If isolated subsegmental thrombi are considered clinically relevant, a negative CT should be followed by an assessment of the deep venous system. (c)1999 by CVRR, Inc. PMID- 11416522 TI - Optimal Cardiac Care at Elderly Age: A Contemporary Challenge. PMID- 11416523 TI - Hospitalization for Heart Failure in the Elderly. AB - People over 65 years account for more than 80% of heart failure hospitalizations, with almost half in patients morer than 75. Heart failure hospitalizations accounted for 21% of the annual health care budget for a representative senior program. Heart failure with preserved ejection fraction accounts for over half of heart failure hospitalizations in the elderly. Current therapy for the elderly is directed to relieve congestion by reducing volume overload and hypertension, while addressing exacerbating factors such as ischemia and anemia. Heart rate reduction is critical when sinus rhythm cannot be maintained but can also improve diastolic filling during sinus rhythm. While cardiac transplantation is rarely indicated, other interventions should be actively considered. Most elderly patients admitted with heart failure wish resuscitation. As heart failure progresses, decisions regarding implantable defibrillators or dialysis require careful consideration, and the risk/benefit balance may shift toward therapies to improve quality of life. (c)1999 by CVRR, Inc. PMID- 11416524 TI - Is Primary Angioplasty the Best Treatment for AMI in the Elderly? AB - The elderly, particularly those over 80, are the fastest growing component of the population. Coronary artery disease accounts for 44% of all deaths in the elderly. Age is also one of the strongest predictors of mortality from acute myocardial infarction (AMI) and in those that survive, long term outcome is worse. In part, the poor inhospital mortality is due to difficulty in diagnosis, but is also likely to be caused by the widespread reluctance to use reperfusion therapy to treat the elderly. Reperfusion therapy has been shown to be equally effective in the elderly, despite an overall increased mortality. The results of randomized trials comparing thrombolysis to angioplasty show an advantage of primary angioplasty over thrombolysis. In the Global Use of Strategies to Open Occluded coronary Arteries in Acute Coronary Syndromes (GUSTO) IIb trial, the advantage was noted in all age groups, including the elderly. The Primary Angioplasty in Myocardial Infarction (PAMI) trial supports these findings. The trials also suggest that the risks are lower with angioplasty than thrombolysis, with a significant lower risk of stroke. Today, with improved outcomes using stents and glycoprotein IIb/IIIa agents, the advantages of primary angioplasty may be even greater than those reported in prior trials. The available data strongly support the use of primary angioplasty in the elderly as an effective reperfusion therapy and, due to improved safety and greater efficacy, should be the preferred treatment in the elderly. (c)1999 by CVRR, Inc. PMID- 11416525 TI - Anticoagulation of the Elderly Patient. AB - The optimal management of various thromboembolic and vascular disorders often entails the use of long term oral anticoagulant therapy. These disorders are generally more prevalent in the elderly than in the young. Hemorrhage is the most common complication of anticoagulant therapy. Although studies have provided conflicting results, it is likely that the elderly are at increased risk of major hemorrhage during prolonged treatment with oral anticoagulants. This review will discuss the reasons why older patients may be at increased risk for anticoagulant related bleeding, the clinical implications of this increased hemorrhagic risk, and strategies for reducing the frequency of major hemorrhage in this growing patient population. (c)1999 by CVRR, Inc. PMID- 11416526 TI - Health Care for the Elderly: Policy Demands and Individual Needs. AB - As with two storm systems colliding, one of the great health care struggles over the next few decades will be the clash between the demands of policy to reform the Medicare program in order to manage its growing costs and the ever improving but usually expensive clinical care that can now be provided to elderly patients. The extension to the elderly of surgical and other forms of treatment once thought impossible, and the antiageist triumph that extension symbolizes, is a remarkable triumph for medical progress. But managing the costs of that triumph has yet to be mastered, portending a rationing of high technology medicine for the elderly in the years to come. Fortunately, improved living standards and public health are likely to see better health for the elderly even if there is a limitation of expensive technologies. (c)1999 by CVRR, Inc. PMID- 11416527 TI - Observations on the Heart - Congenitally Bicuspid Stenotic Aortic Valves in Octogenarians. PMID- 11416528 TI - Images in Geriatric Cardiology - Staphylococcus aureus Mitral Valve Endocarditis. PMID- 11416529 TI - Electrocardiology Teacher Analysis and Review - Acute Inferior Myocardial Infarction With Complications. PMID- 11416530 TI - Ethical Issues in the Management of Geriatric Cardiac Patients - Everyone knew he would not have wanted to live like this.but his advance directive was not clear about his wishes. PMID- 11416531 TI - Hypertension Control at Elderly Age-We Can and Should Do Better. PMID- 11416533 TI - Hypertension in the Aging Patient: New Imperatives, New Options. AB - Over half of all people over the age of 65 in the U.S. have hypertension. In most cases this is diagnosed because of increased systolic blood pressure. It is now recognized that systolic blood pressure is more predictive of cardiovascular events than diastolic blood pressure; since these events are the major cause of death and disability in this population, current hypertension guidelines now emphasize more aggressive blood pressure criteria for both diagnosing and treating systolic hypertension. This process has been stimulated by evidence from large clinical trials that reducing systolic blood pressure improves survival and prevents strokes, heart failure, and other cardiovascular outcomes. The guidelines of both the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC VI) and the World Health Organization-International Society of Hypertension (WHO-ISH) recommend that, regardless of age, hypertension can be diagnosed when the systolic blood pressure is greater than 140 mm Hg. The treatment target is less than 140 mm Hg, though in the presence of concomitant conditions like diabetes mellitus or cardiac or renal impairment, which are common findings in the elderly, even lower target levels may be justified. For patients with systolic blood pressures in the range 140 mm Hg-159 mm Hg but who are without other cardiovascular risk factors, it is not yet certain that aggressive treatment is warranted. New clinical trials are now addressing this question. So far, most experience with treating systolic hypertension in older persons has been with diuretics and calcium channel blockers. But growing evidence indicates that most antihypertensive drug classes are effective and that agents should be selected to best match the needs of individual patients. (c)2000 by CVRR, Inc. PMID- 11416534 TI - Pathophysiology of Hypertension in the Elderly. AB - The onset and progression of hypertension is associated with alterations in structure, function, and hemodynamics of the heart, vascular system, and other major organs. An understanding of the structural and functional changes in the cardiovascular system associated with this process would allow for early detection and the development of treatment strategies. In this paper, we focus on the anatomic alterations that accompany vascular aging and the resulting cardiovascular dynamics. Techniques to measure changes in cardiovascular dynamics and left ventricular performance are reviewed. The impact of therapeutic strategies on cardiovascular dynamics and left ventricular function are also discussed. (c)2000 by CVRR, Inc. PMID- 11416535 TI - The Evidence for Lifestyle Modification in Lowering Blood Pressure in the Elderly. AB - Lifestyle modifications are a safe, effective treatment option for hypertension, yet are widely underutilized by clinicians, particularly in the elderly population. This review outlines the nonpharmacologic approaches to the treatment and prevention of hypertension, and discusses their applicability to the elderly population. Accepted recommendations include weight loss, regular exercise, decreased alcohol consumption, and dietary modifications. Each of these changes has been shown to independently reduce blood pressure in elderly individuals, as well as reduce the incidence and severity of comorbid conditions which potentiate cardiovascular risk. The safety of these recommendations equals or exceeds that of pharmacologic therapy, and few contraindications to their implementation exist. A simple, safe, and effective plan appropriate and acceptable to the geriatric patient is described, offering primary care physicians a guideline to follow when prescribing lifestyle modifications to the elderly hypertensive patient. (c) 2000 by CVRR, Inc. PMID- 11416536 TI - Treatment of Hypertension in the Institutionalized Elderly. AB - The goal of antihypertensive therapy in elderly institutionalized persons is to reduce the blood pressure to less than 140/90 mm Hg if possible. Elderly persons with diastolic hypertension should have their diastolic blood pressure lowered to 80-85 mm Hg. Elderly persons with stage 2 or 3 hypertension, stage 1 hypertension and target organ damage, evidence of clinical cardiovascular disease, or diabetes mellitus should be treated with antihypertensive drug therapy immediately in addition to lifestyle modification. The initial antihypertensive drug in elderly persons without associated medical conditions should be a diuretic or A -blocker because these drugs have been shown to decrease cardiovascular morbidity and mortality in controlled clinical trials. The initial antihypertensive drug in elderly persons should depend on the associated medical condition. If a second antihypertensive drug is indicated, a drug from another class should be administered. If a diuretic is not the initial drug, it is usually indicated as the second drug. If the antihypertensive response is inadequate after reaching the full dose of two classes of drugs, a third drug from another class should be added. Causes of secondary hypertension should be identified and treated. (c)2000 by CVRR, Inc. PMID- 11416537 TI - The Growing Role of Calcium Antagonists in Treating Hypertension in the Elderly. AB - Increasing attention has focused on the magnitude of hypertension in the elderly, with recent data indicating that it afflicts over 50% of people greater than 65 years of age. Recent studies have clearly demonstrated that treatment of isolated systolic and diastolic hypertension in older patients confers substantive protection against hypertension induced morbidity and mortality, to an extent greater than previously appreciated. The results of the recent Systolic Hypertension in Europe (Syst-Eur) trial have demonstrated a striking decrease in the occurrence of strokes, as well as heart failure and all cardiac events in the active treatment group. Because a long acting dihydropyridine calcium antagonist was used in the Syst-Eur trial, it is reasonable and appropriate to recommend a long acting calcium antagonist as one of the preferred agents in the management of isolated systolic hypertensive (ISH) patients. Recent attention has also focused on the importance of formulation and pharmacokinetics as a determinant of cardiovascular risk. It is clear that the newer slow release formulations and intrinsically long acting calcium antagonists are to be preferred. By virtue of their ability to attain more gradual and sustained plasma levels, they do not evoke reactive sympathetic activation. Concomitantly, such formulations should promote increased patient compliance and thereby favorably influence hypertension related morbidity and mortality. (c)2000 by CVRR, Inc. PMID- 11416538 TI - The Need for Continuing Education in Geriatric Cardiology. PMID- 11416539 TI - Hormonal Replacement Therapy for Prevention and Treatment of Atherosclerosis in Postmenopausal Women: A Neglected Aspect of Cardiovascular Education. PMID- 11416540 TI - An Elderly Man With Progressive Dyspnea on Exertion and Atrial Fibrillation as Manifestations of Senile Cardiac Amyloidosis. AB - Senile cardiac amyloidosis is a form of amyloidosis seen more commonly in the elderly population. It was previously believed to have no functional significance, but more recent studies suggest that it is an important cause for cardiovascular morbidity and mortality in the elderly. The diagnosis may be challenging and it is important to have a high suspicion for this disease in the proper clinical setting. A patient with cardiac amyloidosis is presented in this paper to illustrate the clinical context in which one should consider this diagnosis. Information regarding diagnostic evaluation and a brief review of senile cardiac amyloidosis is presented. (c)2000 by CVRR, Inc. PMID- 11416541 TI - Rotational Atherectomy in Octogenarians: Results and Follow Up. AB - We examined our first 390 rotational atherectomy procedures to determine success and complications rates of patients aged 80 or greater (Group I, N equals 35) as compared to those younger (Group II, N equals 355). Procedural success was achieved in 32/35 (91%) of Group I and 42/355 (96%) of Group II patients (p equals NS). Coronary artery bypass graft surgery was required in 2/35 (5.7%) of Group I and 7/355 (2%) of Group II patients. No deaths, Q wave infarctions, or perforations occurred in the elderly patients and only one of each was observed in the younger group. During clinical follow up (26 months, 3-50), 4/25 patients (16%) in the octogenarian group died. Two of the deaths were cardiac (including the only crossover to surgery). One patient who sustained a late non Q myocardial infarction was asymptomatic. Of the remaining 20 patients the anginal class was 0 II in 19 and III in one. Rotational atherectomy can achieve excellent short and long term outcomes in selected octogenarians. (c)2000 by CVRR, Inc. PMID- 11416542 TI - Acute Congestive Heart Failure and Tumor Necrosis Factor Alpha. AB - Congestive heart failure is prevalent in the elderly; recurrent hospital admissions for acute deterioration is a major health care problem. We examined tumor necrosis factor alpha levels in 31 congestive heart failure patients who were admitted to the hospital due to acute cardiogenic pulmonary edema. Blood samples were taken within 24 hours of admission and at the time of discharge after recovery. Tumor necrosis factor alpha levels were also measured in 10 stable congestive heart failure outpatients and 16 healthy controls. Tumor necrosis factor alpha level at admission was higher than at time of discharge (4.6A+/-1.9 vs. 3.4A+/-1.1 (rg/mL, p is less than 0.02). Mean tumor necrosis factor alpha level of stable congestive heart failure outpatient was 2.6A+/-0.7 rg/mL healthy controls had significantly lower tumor necrosis factor alpha levels (1.7A+/-0.7 rg/mL) than congestive heart failure patients (p is less than 0.01). tumor necrosis factor alpha levels were highest among acute congestive heart failure patients. Elevated tumor necrosis factor alpha appears to be related to the exacerbation of congestive heart failure and may be a marker for deterioration. (c) 2000 by CVRR, Inc. PMID- 11416543 TI - CME Paper: Hormone Replacement Therapy for Prevention or Treatment of Atherosclerosis in Postmenopausal Women: Promises, Controversies, and Clinical Trials. AB - Cardiovascular disease is the leading cause of death among men and women in the U.S. and other developed societies. However, myocardial infarction and stroke are uncommon in women until their sixth decade and beyond. Clinicians have long suspected that the delay of a decade or more in cardiovascular disease expression in women relative to men is due to the protective effects of estrogen prior to menopause. Reports from population based observational studies of the favorable effects of hormone replacement therapy on cardiovascular morbidity and mortality have led to enthusiasm for widespread use of estrogen by postmenopausal women for the prevention of cardiovascular disease. In support of the antiatherogenic potential of estrogen are studies in postmenopausal women showing favorable effects on lipoprotein levels, fibrinolysis, and vascular function. However, a secondary prevention trial in postmenopausal women with coronary artery disease showed no cardiovascular benefit of hormone replacement therapy. Ongoing clinical trials and options to conventional hormone replacement therapy for cardiovascular protection will be discussed in this review. (c)2000 by CVRR, Inc. PMID- 11416544 TI - Observations on the Heart - Aneurysm of the False Channel of Descending Thoracic Aorta Years After Operative Excision of the Initiating Aortic Dissection Tear in Ascending Aorta. PMID- 11416545 TI - Images in Geriatric Cardiology: Back Pain and Fatigue. PMID- 11416546 TI - Electrocardiology Teacher Analysis and Review: Hypocalcemia. PMID- 11416547 TI - Ethical Issues in the Management of Geriatric Cardiac Patients: An elderly mute and deaf patient with a known history of cardiac dysrrhythmia is evaluated in the absence of a sign interpreter and subsequently discovered to have been misinterpreted. PMID- 11416548 TI - Cardiovascular Pharmacotherapy at Elderly Age. PMID- 11416550 TI - Cardiovascular Drug Use in the Elderly. AB - The elderly patient is at increased risk for adverse drug effects because of altered pharmacokinetic and pharmacodynamic changes that occur with aging. In addition, the potential for adverse drug interactions are increased because the elderly take a disproportionate number of drugs relative to younger patients. Cardiovascular drugs are frequently implicated in the occurrence of adverse drug interactions. Recommendations are provided to minimize or avoid adverse drug reactions in the elderly. (c) 2000 by CVRR, Inc. PMID- 11416551 TI - Hypertension in the Elderly. AB - Hypertension as a risk factor for cardiac and cerebrovascular morbidity and mortality poses a major health problem for our increasingly elderly population. Recent trials have shown large reductions in stroke, heart failure, and coronary artery disease when elderly hypertensive patients are treated. These benefits are also seen in elderly patients with isolated systolic hypertension. The elderly patient with hypertension should be investigated and managed in a similar manner to their younger counterpart. Nonpharmacological measures, such as dietary salt and calorie restriction, regular exercise, cessation of smoking, and reduction of excess alcohol intake, should be recommended. If these are insufficient, pharmacological treatment should be tailored to the individual patient. Diuretics have been shown to improve outcome measures in the elderly. Other antihypertensive drugs may be added or substituted depending on the patient's blood pressure response to therapy and their comorbid conditions. If all elderly hypertensive patients were treated, a major reduction in cardiovascular and cerebrovascular mortality and morbidity would result. Consideration also needs to be given to nonpharmacological treatment, particularly salt restriction in older subjects where blood pressure is at the upper limit of normal, as this would also result in a major reduction in cardiovascular morbidity and mortality. (c) 2000 by CVRR, Inc. PMID- 11416552 TI - CME Paper: Acute Myocardial Infarction in the Elderly. AB - Up to half of admissions with myocardial infarction are over 70 years of age. Mortality in this age group is higher than in younger age groups. However, elderly patients are less likely to be managed in an acute coronary care setting. Low dose aspirin should be given to all elderly patients with myocardial infarction. Thrombolytic agents have the greatest effect in the elderly even though they give an increased risk of hemorrhagic stroke. They are underused in the elderly with myocardial infarction for a number of reasons. A -Blockers reduce mortality post infarction and ACE inhibitors improve morbidity and mortality rates in those with evidence of heart failure post infarction. Amiodarone may also be of use as an antiarrhythmic in the post infarction period. (c) 2000 by CVRR, Inc. PMID- 11416553 TI - The Management of Chronic Ischemic Heart Disease in the Elderly. AB - Despite advances in medical and surgical therapy, ischemic heart disease remains the most important cause of death and disability in an ever growing population. The management of elderly patients with ischemic heart disease is made more difficult by the presence of comorbid medical conditions, which can both further aggravate ischemic heart disease and interfere with pharmacological and interventional therapy. Despite the difficulties in the management of ischemic heart disease in the elderly, there is substantial evidence that older patients can benefit greatly from aggressive, rational treatment of this condition and, therefore, such treatment should not be denied solely on the grounds of age. (c) 2000 by CVRR, Inc. PMID- 11416554 TI - Arrhythmia Therapy in the Elderly. AB - As with other illnesses, the risks and benefits of diagnostic studies and treatments for arrhythmias are altered by age. In the elderly, the risks of treatment are often greater; drug metabolism varies and mechanical approaches (ablation procedures and insertion of electrical devices) are associated with greater complication rates. This paper reviews recommendations for diagnosis and treatment of arrhythmias in the elderly. (c) 2000 by CVRR, Inc. PMID- 11416555 TI - Nonpharmacologic Management of Supraventricular Tachycardia. AB - This paper reviews three recent large scale studies on the use of catheter ablation for elderly patients with supraventricular tachycardia. These patients underwent catheter ablation of the atrioventricular junction for control of atrial fibrillation, ablation of the slow atrioventricular nodal pathway for those with atrioventricular nodal, reentry tachycardia, and ablation of the accessary pathway for those with atrioventricular reentry tachycardia. There was a higher incidence of atrioventricular junctional ablation and lesser incidence of junctional tachycardia compared to other series involving younger individuals. There was no significant difference in the incidence of successful procedures comparing the elderly with younger patients. In addition, there was no significant difference in the incidence of adverse effects between the two groups. In summary, catheter ablation appears to be safe and effective for elderly patients and the procedure should not be omitted solely on the basis of patient age. (c) 2000 by CVRR, Inc. PMID- 11416556 TI - Exercise Radionuclide Cineangiography for Predicting Long Term Prognosis in Elderly Post Coronary Artery Bypass Graft Patients. AB - ISSUE: The immediate benefits of coronary artery bypass grafting may only be transient. Recent studies which showed that exercise radionuclide cineangiography studies may predict cardiac events and survival had evaluated post coronary artery bypass grafting patients of all ages. This prospective study was done on an elderly subgroup. METHODS: The results of exercise radionuclide cineangiography performed early (4-8 months) post coronary artery bypass grafting were correlated over six years of follow up with mortality, major surgical and nonsurgical cardiac events, and cardiac event free survival in 87 consecutive patients at or above 65 years. RESULTS: The rest to peak exercise difference in left ventricular ejection fraction correlated with later cardiac death and the need for surgical interventions, while rest left ventricular ejection fraction correlated only with the latter parameter. CONCLUSIONS: Assessment of left ventricular ejection fraction at rest and at exercise has prognostic values after coronary artery bypass grafting in elderly patients. Exercise radionuclide cineangiography carried out soon after coronary artery bypass grafting might be considered an useful tool to assess patient prognosis. The efficacy of exercise radionuclide cineangiography carried out soon after coronary artery bypass grafting should be confirmed in larger and more varied patient subgroups. (c) 2000 by CVRR, Inc. PMID- 11416558 TI - Electrocardiology Teacher Analysis and Review - Atrial Flutter. PMID- 11416557 TI - Images in Geriatric Cardiology: Transesophageal Echocardiographic Assessment of Aortic Atheromatous Plaque Morphology in Elderly Stroke Patients. PMID- 11416559 TI - Pharmacologic Update - ACE/Kininase II Inhibitors in Older Patients With Cardiovascular Diseases: "HOPE" Springs Eternal. PMID- 11416560 TI - Clinical Trials: The HOPE Trial. PMID- 11416561 TI - Ethical Issues in the Management of Geriatric Cardiac Patients: A terminal/dying patient endures repeated shocks from an automatic implantable cardioverter defibrillator unit. PMID- 11416562 TI - Cardiovascular Therapies at Elderly Age: New Questions, New Answers. PMID- 11416564 TI - CME Paper: Homocysteine: The New Risk Factor for Cardiovascular Disease in the Elderly. AB - Higher levels of plasma homocysteine are associated with a variety of atherosclerotic conditions that are particularly pertinent to elderly persons. Cross sectional data have suggested that the lower deciles of plasma and dietary folate are highly associated with the presence of carotid stenosis in the elderly. Recent prospective studies have demonstrated strong associations between elevated concentrations of homocysteine and the incidence of cardiovascular mortality, total mortality, and stroke in elderly men and women. The impact of folate fortification on homocysteine levels has been assessed in middle-aged adults, but the clinical impact of fortification or supplementation of the diet with folate on clinical end points of cardiovascular disease is presently unknown, although clinical trials with folate supplementation are underway. (c) 2000 by CVRR, Inc. PMID- 11416565 TI - Early Prognostic Assessment and Treatment of Acute Myocardial Infarction in the Elderly. AB - The progressive aging of the population is associated with an increase in the proportion of very old patients (greater than 75 years) hospitalized with acute myocardial infarction. The lack of evidence regarding the efficacy of most therapeutic interventions for acute myocardial infarction in these patients is leading to a significant degree of uncertainty in the cardiology community with respect to their optimal management. When aggressive treatment (defined as a therapeutic strategy designed to obtain and maintain a patent infarct-related coronary artery at an early moment) of acute myocardial infarction is considered in very old patients, three main questions should be addressed: why should we treat? Whom should we treat? And how should we treat? To answer these questions, the authors reviewed the data available in the literature as well as new data from the PPRIMM75 (PronA3stico del PRimer Infarto de Miocardio en Mayores de 75 aA+/-os) Registry, a large, prospective database of patients aged 75 years or older, admitted to a single coronary care unit in Madrid, Spain, for their first acute myocardial infarction during the last decade. (c) 2000 by CVRR, Inc. PMID- 11416566 TI - Transesophageal Echocardiographic Assessment of Proximal Coronary Artery Stenosis in Elderly Patients With Ischemic Stroke. AB - Coronary artery disease is common in elderly patients with ischemic stroke. Most patients with ischemic stroke undergo transesophageal echocardiography to rule out cardiac sources of cerebral embolism. Transesophageal echocardiography has been shown to be very accurate in diagnosing significant proximal coronary artery stenosis, and may be used to evaluate the coronary arteries for the presence of stenosis in elderly patients with ischemic stroke. Transesophageal echocardiographic detection of these lesions might lead to subsequent coronary angiography and revascularization, dramatically changing the management and outcome of these patients. (c) 2000 by CVRR, Inc. PMID- 11416567 TI - Hormonal and Nonhormonal Therapies for the Postmenopausal Woman: What is the Evidence for Cardioprotection? AB - Despite biologically plausible mechanisms whereby estrogen may confer cardioprotection as well as observational data suggesting cardiovascular benefit, data from the sole randomized controlled clinical outcomes trial reported on the benefit of hormone use. A trend was observed in the Heart and Estrogen/progestin Replacement Study (HERS) of an early increase in coronary events with possible late benefit, and in a recent angiographic trial, the Estrogen Replacement and Atherosclerosis trial (ERA) where no benefit was seen. Furthermore, selective estrogen receptor modulators may enable dissociation of estrogen risks and benefits; the selective estrogen replacement modulator raloxifene is under study in a large randomized clinical outcomes trial entitled the Raloxifene Use for the Heart trial (RUTH). (c) 2000 by CVRR, Inc. PMID- 11416568 TI - Quality of Life and Self Efficacy in Cardiac Rehabilitation Patients Over 70 Years of Age Following Acute Myocardial Infarction and Bypass Revascularization Surgery. AB - Does 12 weeks of cardiac rehabilitation improve quality of life and self efficacy in patients greater than 70 years of age following an acute myocardial infraction or bypass surgery? Three hundred forty four patients were divided into an Older group (70-89 years of age; n equals 122) and a younger group (50-69 years of age; n equals 222). Two hundred fifteen had bypass surgery (Surgical group) and 129 had a myocardial infarction (MI group). Quality of life emotions domain was greater for the Older group at week 12 (p equals 0.066). The Surgical group had greater increases in overall quality of life and quality of life limitations domain (p equals 0.012; p is less than 0.001). Self efficacy scores were greater for the Younger group. MI group had greater self efficacy at week 1, while the Surgical group was greater at week 12 (p is less than 0.001). These results suggest age and diagnosis related differences in quality of life and self efficacy. (c) 2000 by CVRR, Inc. PMID- 11416570 TI - Images in Geriatric Cardiology: Visualization of a Large Mediastinal Mass Compressing the Esophagus and Causing Chest Pain and Dysphagia. PMID- 11416569 TI - Results of Stent Supported Percutaneous Transluminal Coronary Angioplasty in Octogenarians With Coronary Artery Disease. AB - Between January 1995 and July 1998, percutaneous transluminal coronary angioplasty was performed on 27 lesions in 24 octogenarians. Half of the patients were African American. Women comprised 67% of the study group. Patients with unstable angina and myocardial infarction constituted 54% of the cohort. Two thirds of the patients (83%) had single vessel disease with predominant class A and B lesion complexity of the angioplasty site. Acute success rate was 92%. Stents were successfully placed in 11 subjects (46%). None had acute myocardial infarction, emergency coronary artery bypass surgery, or stroke as a complication of the procedure. One patient presenting with acute myocardial infarction complicated by cardiogenic shock, died. Significant bleeding complications requiring blood transfusions occurred in 17% of patients. Of the patients, 23 (96%) were discharged in a clinically stable condition. Follow up during a two year period was completed in 21 patients (88%). One patient died of cancer. Four subjects (19%) underwent repeat percutaneous transluminal coronary angioplasty. One other patient had recurrent chest pain requiring multiple hospitalizations. The remaining 16 patients (76%) remained free of recurrence of angina. We concluded that percutaneous transluminal coronary angioplasty with stent placement can be performed in octogenarians with a high rate of clinical and angiographic success with an acceptable range of morbidity and mortality, and favorable long term (two year) outcome. (c) 2000 by CVRR, Inc. PMID- 11416571 TI - Electrocardiology Teacher Analysis and Review: Wenckebach (Mobitz I) Atrioventricular Block. PMID- 11416572 TI - Observations on the Heart: Wide Open Coronary Arteries at 103 Years of Age. PMID- 11416573 TI - Ethical Issues in the Management of Geriatric Cardiac Patients: A terminally ill patient changes his mind and opts for futile intervention after discovering that his insurance will pay for it. PMID- 11416574 TI - Cardiovascular Research in the Elderly. PMID- 11416575 TI - Executive Summary: Pivotal Research in Cardiovascular Syndromes in the Elderly. AB - The PRICE-1 conference was designed to identify near term priorities for funding cardiovascular research in the elderly. Twenty topics were identified with either break throughs in fundamental mechanisms of aging with cardiovascular systems or with critical importaance to cardiovascular carve of the elderly. (c) 2000 by CVRR, Inc. PMID- 11416576 TI - Research Agenda for Cardiovascular Aging: Humans to Molecules. AB - Clinical manifestations of specific cardiovascular diseases, e.g., atherosclerosis and hypertension, that lead to heart failure and stroke likely become altered in older persons of advanced age because interactions occur between age-associated cardiovascular changes in health and specific pathophysiologic mechanisms that underlie cardiovascular diseases. The interactions result in a lower threshold for clinical symptoms, and greater severity and poorer prognosis of these diseases in older vs. younger persons. In this regard, cardiovascular changes that occur during aging in health ought not to be considered to reflect a "normal process"; rather these specific age associated changes must be construed as specific "risk factors" for the aforementioned cardiovascular diseases and ought to become targets of interventions designed to prevent the epidemic of cardiovascular disease in later life. Such a strategy would thus advocate preventive treatment for what is now considered to be "normal cardiovascular aging." Effective and efficient prevention of the "risks" associated with cardiovascular aging in apparent health requires a fundamental understanding of these changes ranging in scope from humans to molecules. An opinion regarding specific directions for research aimed toward the achievement of this understanding is provided. (c) 2000 by CVRR, Inc. PMID- 11416577 TI - Apoptosis in Cardiovascular Aging Research: Future Directions. AB - Apoptosis is the process by which cells are induced to activate their own suicide. Programmed cell death occurs in a wide variety of cell types, including cardiovascular tissues. There is increasing evidence of a relationship between apoptosis and cardiovascular disease, particularly ischemic heart disease and congestive heart failure, the most frequent heart diseases in the older population. Research directed to the treatment and possible prevention of apoptosis may provide a means of decreasing the incidence of cardiac failure and increasing the survival of endothelial and smooth muscle cells in the elderly. (c) 2000 by CVRR, Inc. PMID- 11416578 TI - Elderly Patients with Acute Coronary Syndromes: Higher Risk and Greater Benefit from Antithrombotic and Interventional Therapies. AB - In elderly patients with acute coronary syndromes, outcomes are poorer than in younger patients and, disappointingly, some therapies-including thrombolysis for ST elevation myocardial infarction-confer less benefit than in younger patients. In contrast, in unstable angina and non-ST elevation myocardial infarction, the elderly appear to derive greater relative and absolute benefit from the newer, more potent antithrombotic therapies. In both the Efficacy and Safety of Subcutaneous Enoxaparin in Non-Q-Wave Coronary Events and Thrombolysis in Myocardial Infarction 11B trials, the low molecular weight heparin enoxaparin, compared with unfractionated heparin, appeared to have greater relative and absolute benefit in patients aged 65 years and older, as compared with younger patients. For the glycoprotein IIb/IIIa inhibitors, an equivalent relative benefit has been observed, which translated into a greater absolute benefit in older vs. younger patients. Similarly, in the FRagmin and Fast Revascularisation during InStability in Coronary Artery Disease II trial, patients 65 years and older derived significantly greater benefit from an invasive than from a conservative strategy, whereas there was no difference in outcome by strategy in younger patients. A similar trend was observed in the Thrombolysis in Myocardial Infarction IIIB trial. Thus, in unstable angina and non-ST elevation myocardial infarction, elderly patients are at higher risk and appear to derive particular benefit from the more aggressive antithrombotic and interventional therapies. (c) 2000 by CVRR, Inc. PMID- 11416579 TI - Ischemic Heart Disease in the Elderly:A Need to Understand the Causes of High Mortality After Acute Myocardial Infarction. AB - The short- and long-term mortality of patients greater than 75 years of age with acute myocardial infarction is still very high. These patients have been excluded from most randomized controlled trials designed to test interventions directed to improve the outcome of acute myocardial infarction. Contrary to young-old patients (65-74 years) in whom the benefit obtained with these interventions is even greater than that observed in younger patients, there is a lack of information concerning the optimal treatment of acute myocardial infarction in the oldest patients. Research specifically directed to assess the optimal reperfusion strategy and coadjuvant therapies in patients greater than 75 years old is critically important. The causes for such high mortality are still poorly understood. Several clinical observations suggest that old patients with acute myocardial infarction have specific pathophysiologic behaviors. As the knowledge of the mechanisms of disease is, in clinical practice, the basis of therapy development, clinical and basic research designed to elucidate the specific pathophysiological mechanisms and causes of the high mortality in the oldest patients with acute myocardial infarction should be regarded as a priority in geriatric cardiovascular research. (c) 2000 by CVRR, Inc. PMID- 11416580 TI - Orthostatic Hypotension in the Elderly: Contributions of Impaired LV Filling and Altered Sympathovagal Balance. AB - Orthostatic hypotension, which occurs in 5%-18% of the elderly, may contribute to age-related disability. While autonomic dysfunction and alterations of cardiac structure and function likely to impair postural maintenance of blood pressure are common in the elderly, these have not been jointly studied in large cohorts. The authors evaluated the association of orthostatic hypotension with echocardiographic measures of cardiac structure and function, and with autonomic function determined by analysis of heart rate variability, in a large population of community-dwelling elderly. A total of 5201 men and women, aged 65-100 years and living in four geographically separate communities, were recruited from Medicare eligibility lists. In this prospective, observational cohort study, measurements included clinical questionnaires, standing and supine blood pressures, mini-glucose tolerance testing, echocardiography, and 24-hour Holter recording for assessment of heart rate variability. Orthostatic hypotension, defined as a decrease in standing systolic blood pressure of 20 mm Hg or more, was positively associated in bivariate analyses with left ventricular wall thickness, peak velocity of late diastolic filling, vagal tone on heart rate variability analysis, supine systolic pressure, supine diastolic pressure, age, and diabetes, and inversely associated with body weight. After statistical adjustment for the presence of myocardial infarction, stroke, and use of antihypertensive medication, the associations were maintained, and a previous trend toward an association with decreased left ventricular cavity size became statistically significant. The data suggest that in elderly, community-based individuals, orthostatic hypotension is associated with increased blood pressure and decreased weight; it possibly acts mechanistically via altered sympathovagal balance, increased left ventricular wall thickness, decreased left ventricular preload, and alterations of left ventricular diastolic filling. (c) 2000 by CVRR, Inc. PMID- 11416581 TI - CME Paper: Lipid Lowering Therapy is Safe in the Elderly:The LifeHelp 80-80 Analysis. AB - Little information is available regarding the safety and efficacy of lipid lowering therapy in the elderly. Records of 80 octogenarians, taken from the database of the LifeHelp Lipid Clinic of the Heart Institute of St. Petersburg, FL, were examined, representing 1863 patient months of treatment. Therapy was effective, lowering low density lipoprotein cholesterol (from 140A+/-4 to 92A+/-3 mg/dL), raising high density lipoprotein cholesterol (from 44A+/-1 to 50A+/-1 mg/dL), and lowering triglycerides (from 199A+/-21 to 146A+/-9 mg/dL). Six patients developed myalgias (without significant creatine phosphokinase rise) or gastrointestinal disturbances. Three of these patients restarted therapy without difficulty. No patient developed rhabdomyolysis (including the 11% of patients on combination therapy). One patient developed an elevation of liver (more than 3 times upper limits of normal), but later restarted therapy without sequelae. We conclude that lipid lowering therapy was effective at modifying hyperlipidemia in the elderly without significant adverse effects. (c) 2000 by CVRR, Inc. PMID- 11416583 TI - Electrocardiology Teacher Analysis and Review: High Grade Atrioventricular Block with an Ectopic Atrial Rhythm. PMID- 11416582 TI - Images in Geriatric Cardiology: Transesophageal 3-D Color Doppler Echocardiographic Findings in Traumatic Aortic Isthmus Pseudoaneurysm. PMID- 11416584 TI - Pharmacologic Update - Beta Blockers in Treatment of Heart Failure and Hypertension in Diabetes: Ugly Misconceptions Slain by Beautiful Clinical Data. PMID- 11416585 TI - Clinical Trial: Risk of Thrombolysis in the Elderly. PMID- 11416586 TI - Ethical Issues in the Management of Geriatric Cardiac Patients: A medically impaired patient is determined to operate an automobile against his physicians' advice. PMID- 11416587 TI - A New Hypertension Journal for the Practicing Physician. PMID- 11416588 TI - Public Health Will be the Winner if the Editorial Goals of This New Journal Are Achieved. PMID- 11416589 TI - Self Reported Sexual Dysfunction in Men and Women Treated With Bisoprolol, Hydrochlorothiazide, Enalapril, Amlodipine, Placebo, or Bisoprolol/Hydrochlorothiazide. AB - Quality of life may be impaired by antihypertensive therapy. Perceived sexual dysfunction by antihypertensive drugs diminishes quality of life and results in noncompliance with antihypertensive therapy. To assess the impact of various classes of antihypertensive therapy vs. combination therapy, self reported adverse reactions were catalogued by gender using COSTART (Coding Symbols for Thesaurus of Adverse Reaction Terms) of impotence or libido decrease in six randomized, blinded, prospective trials in which subjects received placebo, 5 mg qd-20 mg bid enalapril, 2.5-10 mg qd amlodipine, 6.25-25 mg qd hydrocholorothiazide (HCTZ), bisoprolol 5 mg qd, or a combination of 2.5-10 mg qd bisoprolol/6.25 mg HCTZ. The average duration of drug exposure was 6-14 weeks (range of 1 day to 23 weeks). Comparison among groups was performed using Fisher's exact test. There was no statistical difference between treatment with respect to impotence (p equals 0.688), decrease in libido (p equals 0.970), or overall sexual dysfunction (p equals 0.705) for 1251 men. Of the 661 women studied, decrease in libido was reported in only two subjects. It is concluded that short term exposure to antihypertensive drugs is associated with self reported impotence at no greater prevalence than it is with placebo in men. The combination of bisoprolol/6.25 mg HCTZ is not more likely to be associated with sexual dysfunction than placebo, HCTZ, bisoprolol, enalapril, or amlodipine. Also sexual dysfunction is reported less frequently in women than men. (c)1999 by Le Jacq Communications, Inc. PMID- 11416590 TI - Obesity Hypertension in the Atherosclerosis Risk in Communities Cohort: Implications of Obesity Guidelines. AB - An estimated 55% of the U.S. adults are overweight or obese (body mass index [BMI] equals 25 kg/m2). Overweight individuals have a threefold increased risk for the development of hypertension compared to lean individuals. The National Institutes of Health, National Heart, Lung, and Blood Institute (NHLBI) guidelines for treatment of overweight and obese adults recommend weight reduction strategies including pharmacologic treatment with antiobesity agents approved by the FDA. Treatment is recommended for obese individuals and for overweight persons with other risk factors for cardiovascular disease (CVD) including hypertension. This analysis of the Atherosclerosis Risk In Communities (ARIC) cohort for overweight/ obese hypertensive participants indicate that 64% of the hypertensive participants in ARIC deserve consideration for treatment with antiobesity agents according to the current NHLBI guidelines. Thus far there are no long term morbidity and mortality clinical trials to determine the safety of antiobesity agents currently approved by the FDA. The authors caution health care providers in the use of these agents in the obese patient with hypertension. (c)1999 by Le Jacq Communications, Inc. PMID- 11416591 TI - 24 Hour Ambulatory Blood Pressure Monitoring: Is it Necessary to Establish a Diagnosis Before Instituting Treatment of Hypertension? AB - 24 hour ambulatory blood pressure monitoring (ABPM) has been available for many years as a research tool, and is gradually finding its way into clinical practice, despite a general lack of reimbursement in the U.S. Although clinic readings remain the mainstay of blood pressure measurement, they may misrepresent the true blood pressure in a substantial number of patients. Reasons for this include poor measurement technique, the small number of readings that can be taken, and the white coat effect. Both home monitoring and ABPM can overcome these limitations: the former is superior for assessing changes of blood pressure over time, and the latter for assessing the true blood pressure. The major clinical indications for ABPM include newly diagnosed hypertensives in whom there is no target organ damage, patients with suspected white coat hypertension, and some patients with resistant hypertension. Home monitoring may be used as an initial screening test in these patients. (c)1999 by Le Jacq Communications, Inc. PMID- 11416592 TI - Hypertension in Type II Diabetes: Update on Therapy. AB - Cardiovascular disease (CVD) is the major cause of mortality in persons with type II diabetes mellitus and many factors contribute to its high prevalence; hypertension is one such factor. High blood pressure is about twice as frequent in persons with diabetes mellitus as those without the disease. Information from death certificates indicates that hypertension was implicated in 4.4% of deaths coded to diabetes, and diabetes was involved in 10% of deaths coded to hypertension related disease. Up to 75% of diabetes related cardiovascular complications may be attributable to hypertension. These observations have contributed to recommendations for more aggressive lowering of blood pressure (i.e., to less than 130/85 mm Hg) in persons with coexistent diabetes and hypertension. The goal of lowering blood pressure in persons with diabetes is to prevent hypertension associated death and disability. In older persons, the level of blood pressure and the diagnosis of hypertension should be based on multiple blood pressure measurements obtained in a standardized fashion on at least three occasions. Because of the propensity to orthostatic hypotension, standing blood pressures should be measured on patients on each office visit. Further, because of the increased blood pressure variability of these patients, ambulatory blood pressure measurements or home blood pressure monitoring may be valuable. The consensus blood pressure goal in diabetic persons with hypertension is less than 130/85 mm Hg. Pharmacologic therapy should be initiated when lifestyle modifications do not lower blood pressure to less than 130/85 mm Hg in diabetic persons. The National Institutes of Health Consensus Panel recommended four classes of drugs that are effective as first line, single agent therapy. Each drug class has potential advantages and disadvantages but combination therapy is usually necessary for adequate blood pressure control. (c)1999 by Le Jacq Communications, Inc. PMID- 11416593 TI - World Health Organization-International Society of Hypertension Guidelines for the Management of Hypertension-Do These Differ From the U.S. Recommendations? Which Guidelines Should the Practicing Physician Follow? AB - The recent World Health Organization-International Society of Hypertension (WHO ISH) recommendations for the treatment of hypertension are consistent with the guidelines established by the Sixth Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC VI) in the U.S. with several exceptions. Both reports define hypertension as a persistent elevation of blood pressure greater than 140/90 mm Hg and advocate the lowering of blood pressure for all patients with cardiovascular (CV) risk factors in addition to hypertension. The WHO-ISH report, however, suggests continuing monitoring without medication for subjects without other risk factors if pressures are not greater than 150/95 mm Hg. The JNC VI recommends drug therapy even in these subjects if blood pressures remain greater than 140/90 mm Hg after a 6-12 month period of nonpharmacologic interventions. Based on available data this would appear to be a more reasonable recommendation. The WHO-ISH indicates that all classes of medication are suitable initial therapy, despite the lack of morbidity and mortality data with several of them. The JNC VI continues to use outcome data to recommend diuretics or A -blockers as initial treatment. The WHO ISH recommendations may prove to be appropriate-i.e., the lowering of blood pressure makes the difference and not which medication is used-There are some data to support this position but at present the strongest outcome data support the JNC VI recommendations. Both reports stress the importance of lowering blood pressure to levels of 130/85 mm Hg or even lower in patients with diabetes, renal, or heart failure. The addition of the WHO-ISH report should help to focus more clearly the need for better blood pressure control. (c)1999 by Le Jacq Communications, Inc. PMID- 11416594 TI - Hypertension in Older Persons. AB - Hypertension, especially that characterized by elevated systolic blood pressure (SBP), greatly increases cardiovascular risk in older persons; controlled trials have demonstrated that antihypertensive drug treatment is effective in lowering cardiovascular events and mortality in this population. Lifestyle modifications may be effective in reducing blood pressure (BP), but medication is usually necessary in the majority of patients. The goal of therapy is a BP of less than 140/90 mm Hg for most patients and less than 130/85 mm Hg for diabetics. Achieving these levels is, however, difficult in many patients over the age of 65 years. In most older hypertensive patients, low dose diuretics or diuretic based therapy should be considered as initial treatment. Moderately long acting or long acting dihydropyridine calcium antagonists are appropriate in isolated systolic hypertension (ISH) for patients in whom a diuretic is ineffective or poorly tolerated. ACE inhibitors, A -blockers, and diuretics will significantly reduce cardiovascular events in older hypertensive diabetic patients. Multiple drug therapy is necessary in the majority of patients. Despite the availability of effective therapy, fewer than 25% of older hypertensive patients are controlled. (c)1999 by Le Jacq Communications, Inc. PMID- 11416595 TI - Strategies for the Treatment of Hypertension in Postmenopausal Women. AB - Coronary heart disease (CHD) remains the leading cause of death in women. Although premenopausal women experience less CHD than men, mortality from CHD increases more rapidly in women over the age of 60. There are also significant racial differences in mortality among women. Some of these differences can be attributed to genetics, but there are fundamental differences in practice patterns which might contribute to inequality. Coincident with the recent increase in CHD mortality is a growing failure to adequately treat hypertension. The current guidelines for the treatment of hypertension are gender neutral. However, this approach does not account for differences in side effects or treatment failure rates, which vary by gender. It also does not account for gender specific physiologic differences in susceptibility to CHD. This review summarizes the clinical profile of hypertension in women, the rationale for treatment, and the consequences of therapy, with an emphasis on age and gender biased risk assessment. The results of recent trials of antihypertensive therapy in postmenopausal women are reviewed. (c)1999 by Le Jacq Communications, Inc. PMID- 11416596 TI - Current Concepts of Pharmacotherapy in Hypertension: Clinical Implications of the Dose Response Relationship of Antihypertensive Medication in the Treatment of Hypertension. PMID- 11416597 TI - Analysis of Recent Papers in Hypertension. PMID- 11416599 TI - Hypertension Perspectives of the Primary Care Physician: ".More Difficult to Treat Than People Believe." PMID- 11416598 TI - We Can Do a Better Job of Controlling Hypertension. PMID- 11416600 TI - Exercise Training and Blood Pressure Control in Patients With Hypertension. AB - Several randomized, well controlled exercise intervention studies have shown that regularly performed aerobic exercise significantly lowers blood pressure in patients with essential hypertension. Mild to moderate intensity exercise may be more effective in lowering blood pressure than higher intensity exercises. Three or more sessions per week of 30-60 minutes per session are required to achieve optimal results. Such exercise is also safe and effective in lowering blood pressure in treated patients with severe hypertension and left ventricular hypertrophy (LVH). A significant reduction in blood pressure and LVH regression may be achieved in these patients even with substantial reduction in antihypertensive medication. The safety and efficacy of mild to moderate exercise has positive clinical implications for all hypertensive patients. Exercise induced reductions in resting blood pressure and the prevention of an abnormal rise in blood pressure during physical exertion can lead to fewer cardiovascular events. It may also reduce antihypertensive medication requirements, cost, medication related side effects, and improve quality of life. (c)1999 by Le Jacq Communications, Inc. PMID- 11416601 TI - Study of Hypertensive Prescribing Practices (SHyPP): A National Survey of Primary Care Physicians. AB - BACKGROUND: Approximately 50 million people in the U.S. have hypertension. The Joint National Committee (JNC V) guidelines provide treatment recommendations for hypertension. These guidelines promote initiating therapy with diuretics and A blockers because these agents have been shown to reduce hypertension related morbidity and mortality. OBJECTIVES: To survey primary care physicians' antihypertensive prescribing practices, their perceptions about antihypertensive medication cost and safety, and their perceptions of the effects of managed care. DESIGN: A direct mail survey was distributed to a national random sample of 500 office based, primary care internists, family practitioners, and general practitioners. RESULTS: 72% of the physicians who responded reported being aware of the JNC V guidelines. Younger physicians claimed to be more aware of and more likely to follow JNC V guidelines than older physicians. Only 25% of physicians reported initiating treatment at systolic blood pressures greater than 140 mm Hg as recommended by the JNC V guidelines. Most physicians reported initiating treatment at higher pressures. Diuretics were selected for initial antihypertensive treatment by 36% of physicians, ACE inhibitors by 35%, A blockers by 16%, and calcium channel blockers (CCBs) by 7%. Up to 27% of physicians admitted not knowing the cost of specific medications. Approximately half of the physicians reported that they and their patients were more concerned about medication cost and safety today than 18 months ago. A greater number of younger physicians reported that managed care formularies infringed upon their prescribing practices. CONCLUSIONS: Antihypertensive prescribing patterns are inconsistent with JNC V guidelines. Actual practice may deviate even more from these guidelines than reported in this survey. Also, physicians perceive that managed care is increasingly affecting their prescribing practices. (c)1999 by Le Jacq Communications, Inc. PMID- 11416602 TI - The Importance of Moderate Sodium Reduction in the Nonpharmacological Therapy of Hypertension. AB - There are many lifestyle modifications recommended for the prevention and treatment of hypertension. The only modification that has been challenged is the moderate reduction of dietary sodium. As is detailed in this paper, the basis for this challenge are specious and the evidence favoring such moderate sodium reduction are so strong as to recommend it universally. The goal can be reached easily by voluntary removal of some of the excess salt added to processed foods and beverages. (c)1999 by Le Jacq Communications, Inc. PMID- 11416603 TI - Out of Office Self Measurement of Blood Pressure. AB - Self measurement of blood pressure is increasingly common among hypertensive patients. These out of office measurements are clinically useful in avoiding misdiagnosis of hypertension, over treatment and under treatment of hypertension, and as a measure to increase compliance and reduce the cost of antihypertensive therapy. This paper will address the clinical relevance of out of office blood pressure measurement related to hypertension management, the instruments available to perform out of office blood pressure measurement, and the current problems of interpretation of these blood pressure values as they relate to subsequent development of hypertension derived cardiovascular disease. (c)1999 by Le Jacq Communications, Inc. PMID- 11416604 TI - Evaluation of Hypertension in Older Children and Adolescents. AB - Hypertension occurs in children as well as adults. Unlike adults, the rates of hypertension in childhood are lower and the methods of diagnosis and evaluation vary somewhat. In the following review on the diagnosis and management of hypertension in older children, three typical cases will be discussed. Key issues in the methods and criteria used in the diagnosis of hypertension will be described. The secondary causes of hypertension in the young, the characteristics of essential hypertension, and the management of hypertension in the young will also be discussed in the context of typical cases. (c)1999 by Le Jacq Communications, Inc. PMID- 11416605 TI - Treatment of Hypertension and the J Curve. AB - The optimal reduction in blood pressure when treating hypertension is obviously the one which reduces hypertension related cardiovascular morbidity and mortality as much as possible. Today's approach to the treatment of high blood pressure usually does not achieve this goal. In other words, treated hypertensive patients are still at increased risk of cardiovascular complications, such as strokes and myocardial infarcts, in spite of being treated with antihypertensive drugs. One possible explanation for this could be that the treated blood pressure rarely is brought down to strictly normotensive levels, as shown in numerous studies. This would suggest that a more vigorous lowering of blood pressure would provide greater benefit when treating hypertensive patients. On the other hand, the fact that the relationship between the level of blood pressure and risk may be J shaped suggests that vigorous lowering of blood pressure may increase cardiovascular risks, if blood pressure is brought down too far. A few open, retrospective, nonrandomized studies and one randomized placebo controlled study in the elderly (SHEP) have analyzed this issue. The Hypertension Optimal Treatment (HOT) study,1,2 which was published in June 1998,3 has addressed this problem in a prospective randomized trial. The discussions around the J curve with some emphasis on the HOT study will be briefly reviewed. (c)1999 by Le Jacq Communications, Inc. PMID- 11416607 TI - Current Concepts of Pharmacotherapy in Hypertension - Diuretics: Where Have We Been and Where Are We Going. PMID- 11416606 TI - Maximizing Cardiorenal Benefit in the Management of Hypertension: Achieve Blood Pressure Goals. AB - Current recommendations by the Sixth Report of Joint National Committee (JNC) suggest that lower blood pressure levels than previously recommended should be obtained in order to preserve renal function and reduce cardiovascular risk. This holds true for people with diabetes as well as those with renal insufficiency. By all accounts, we as a nation are doing a poor job of even achieving the general guideline for blood pressure control (i.e., less than 140/90 mm Hg). This has contributed to a plateau in mortality reduction from cardiovascular events and a may have contributed to an increase in the incidence of end stage renal disease. This paper reviews the rationale for achieving lower blood pressure goals (i.e., less than 130/85 mm Hg) in people with diabetes and/or renal disease. It also provides an approach as to how to achieve these lower goal blood pressures with a minimum of side effects. Lastly, the use of fixed dose combinations is discussed as a method to improve compliance. In brief, patients with diabetes or renal disease will require an average of 2.6-4.3 different antihypertensive medications to achieve these lower blood pressure targets. The number of medications is a function of the degree of renal insufficiency. If done appropriately, patients can consume these medications in 2 to maybe 3 pills per day. By achieving these lower blood pressure goals there is a good possibility of reducing morbidity as well as mortality from cardiovascular and renal disease. More recent reports document that it is more cost effective in these high risk groups to achieve levels of diastolic blood pressure of less than 85 mm Hg. (c)1999 by Le Jacq Communications, Inc. PMID- 11416608 TI - Common Questions and Answers in the Management of Hypertension - Everyday Care of the Hypertensive Patient: Dental Visits. PMID- 11416609 TI - Analysis of Recent Papers in Hypertension. PMID- 11416611 TI - Lifestyle Interventions: They Lower Blood Pressure: Can We Make Them Work? PMID- 11416610 TI - Low Dose Combination Therapy vs. High Dose Monotherapy in the Management of Hypertension. AB - A common approach to the management of hypertension suggests the use of an initial drug and the addition of a second agent if goal blood pressures (BPs) are not achieved. The Sixth Report of the Joint National Committee (JNC VI) suggests that the use of low dose combination therapy may be an appropriate alternative to initial treatment. In this prospective, randomized, open label, blinded end point (PROBE) study, following a 2 week single blind placebo washout period, qualified patients were started on a calcium channel blocker, felodipine 5 mg for 2 weeks. Patients who were nonresponders (DBP equals 90 mm Hg) were randomized to either felodipine 10 mg (n equals 17, mean age 52.4, 12 males), an ACE inhibitor, enalapril 5 mg/felodipine 5 mg (n equals 20 mean, mean age 52.1, 13 males), or another ACE inhibitor, benazepril 10 mg/amlodipine 5 mg (n equals 18, mean age 53.9, 15 males) for an additional 6 weeks. Ambulatory BP monitoring was performed at the end of the single blind placebo washout period and again at the end of the study. All three treatment groups had significant reductions in mean 24 hour systolic and diastolic BP compared to baseline. The reduction in mean 24 hour systolic and diastolic BPs in the benazepril/amlodipine treated patients (-17.2/ 9.7 mm Hg) was significantly greater (p equals 0.05) than in the felodipine 10 mg treated patients (-11.9/-6.4 mm Hg) and was also numerically but not statistically significantly greater than in the enalapril/felodipine treated patients (-14.2/-8.2 mm Hg). Similar differences were seen when assessing daytime BP (06:00-21:59) and nighttime BP (10:00-05:59). The reductions in morning systolic and diastolic BP (6 am-noon) in the benazepril/amlodipine treated patients (-18.6/-10.7 mm Hg) were numerically but not statistically significantly greater than the enalapril/felodipine treated patients (-13.6/-7.5 mm Hg) and the felodipine 10 mg treated patients (-14.9/-8.3 mm Hg). The data from this study suggests that using low dose combination therapy in patients who are nonresponders to first line monotherapy with a calcium channel blocker provides greater blood pressure control than up titration to higher dose monotherapy. Low dose combination therapy may therefore be an important early alternative to up titration of monotherapy in patients who are nonresponders. These data confirm other observations with combination therapy with A -blockers/ diuretics, ACE inhibitors/diuretics, and angiotensin II (AII) receptor blockers/diuretics. (c)1999 by Le Jacq Communications, Inc. PMID- 11416612 TI - Lifestyle Changes That Reduce Blood Pressure: Implementation in Clinical Practice. AB - Lifestyle modification is widely advocated as initial and adjuvant therapy for the treatment and prevention of hypertension. In addition to exercise, nutritional changes including sodium reduction, adoption of the Dietary Approaches to Stop Hypertension (DASH) diet, alcohol reduction, and weight loss have significant blood pressure lowering effects. The current challenge to clinicians has been developing and implementing practical and effective approaches for use in clinical practice. This review discusses the evidence supporting each current lifestyle recommendation, discusses evidence on specific programs or office based strategies, and provides practical advice. (c)1999 by Le Jacq Communications, Inc. PMID- 11416613 TI - How Low Should We Treat Blood Pressure and Why? AB - With nearly 50 million Americans having high blood pressure, defined as a blood pressure of greater than or equal to 140/90 mm Hg, any therapeutic approach that improves the likelihood of success by either improved tolerability or better capability of keeping blood pressure controlled can have a profound impact on subsequent risk for cardiovascular disease. Therapeutic strategies thus far have not been entirely successful in controlling blood pressure, even to goals of less than 140/90 mm Hg in part due to the asymptomatic nature of the disease process. But maybe this goal should be lowered. Perhaps if target pressures are set at lower levels more people will be treated and outcome improved especially in high risk subjects. About one-half of Americans with high blood pressure are treated. Of those who are treated, only about 50% are controlled. These statistics may explain why treated hypertensives have not had the expected reduction in cardiovascular disease. Consequently, a careful reevaluation of target blood pressures and how to achieve this goal is of great interest. Many questions concerning whether to treat the systolic or diastolic pressures, how low to reduce them, and with which therapy remain unanswered. Specific antihypertensive medications may provide advantages not only with regard to their ability to control blood pressure but also through blood pressure independent mechanisms in preventing vascular and target organ damage. Clinical trials evaluating the benefits of lower target blood pressures with different types of medications are underway and may help provide insight with regard to future strategies to reduce cardiovascular risks in hypertensive patients. (c)1999 by Le Jacq Communications, Inc. PMID- 11416614 TI - Prevalence and Treatment of Hypertension Complicated by Comorbid Conditions. AB - Although the presence of a comorbid condition should be an important factor in etermining the choice of an antihypertensive agent, the prevalence of conditions complicating hypertension is unknown in most patient populations. It is also unknown whether the presence of comorbid conditions influences antihypertensive therapy. To address these questions, we determined the prevalence of comorbid conditions and antihypertensive prescribing patterns for 7526 patients receiving antihypertensive medication at four Department of Veterans Affairs clinics. Of these patients, 97% were male and 86% were over 50 years of age. In these hypertensives, 2507 had coronary artery disease (CAD), 2426 had diabetes mellitus (DM), 1553 had chronic obstructive pulmonary disease (COPD) or asthma, and 970 had congestive heart failure (CHF). The presence of comorbid conditions did not have a major impact on the choice of antihypertensive agents, with most hypertensives receiving calcium antagonists or ACE inhibitors. The use of A blockers, for example, was not greatly influenced by the presence of CAD. (c)1999 by Le JacqCommunications, Inc. PMID- 11416615 TI - Cardiovascular Effects of Cocaine: Focus on Hypertension. AB - Cocaine, a naturally occurring alkaloid, has increasingly been implicated in a myriad of medical complications. The majority of these relate to cardiovascular effects of the drug, a potent sympathomimetic. In addition, cocaine has effects on endothelin-1, the sodium channel, and nitric oxide which further enhance its untoward cardiovascular effects. The cardiovascular effects of cocaine include myocardial ischemia or infarction, ventricular arrhythmias and sudden death, cardiomyopathy, cerebral infarction or hemorrhage, and acute hypertension. Although hypertension has been described in the offspring of cocaine using mothers, two recent studies have not found an increased prevalence of chronic hypertension in adults. Nonetheless, long term abuse of cocaine can lead to the various forms of target organ damage usually associated with untreated essential hypertension, presumably due to frequent intermittent and severe elevations in blood pressure. (c)1999 by Le Jacq Communications, Inc. PMID- 11416616 TI - Current Concepts of Pharmacotherapy in Hypertension - ACE Inhibitors: Current Considerations in Their Use. PMID- 11416617 TI - Common Questions and Answers in the Management of Hypertension - Everyday Care of the Hypertensive Patient: Postmenopausal Estrogen and Blood Pressure. PMID- 11416618 TI - Analysis of Recent Papers in Hypertension. PMID- 11416619 TI - In the Management of Hypertension is it Blood Pressure Lowering Alone or Specific Effects of Specific Medications That Reduce Cardiovascular Events? PMID- 11416620 TI - Nonsustained Ventricular Tachycardia as a Predictor of Cardiovascular Events in Black Men With Hypertensive Left Ventricular Hypertrophy. AB - OBJECTIVE: To determine the prognostic significance of asymptomatic nonsustained ventricular tachycardia in Black males with hypertensive left ventricular hypertrophy without apparent coronary artery disease. BACKGROUND: The Framingham study has demonstrated a higher incidence of cardiovascular mortality including sudden cardiac death in patients with hypertensive left ventricular hypertrophy. Several studies have demonstrated an increased incidence of complex ventricular arrhythmias in patients with left ventricular hypertrophy even in the absence of significant coronary artery disease. There are few long term data, however, regarding the cardiovascular morbidity and mortality associated with asymptomatic ventricular arrhythmias in Black males with essential hypertension and left ventricular hypertrophy. METHODS: Baseline 48 hour Holter monitoring was performed in 110 Black males with hypertensive left ventricular hypertrophy and the presence of nonsustained ventricular tachycardia; the occurrence and frequency of ventricular ectopy were recorded. The study end point was all cause mortality and cardiovascular morbidity (cerebrovascular accident, congestive heart failure, myocardial infarction, and new onset angina). RESULTS: In the 22 patients with nonsustained ventricular tachycardia there were a total of nine events compared to eight events in the 78 patients without nonsustained ventricular tachycardia. During a mean follow up period of 6.4A+/-2.1 years, patients with hypertensive left ventricular hypertrophy and nonsustained ventricular tachycardia on baseline Holter electrocardiogram had an event rate of 41% compared with only 10% in patients without nonsustained ventricular tachycardia (p is less than 0.0001). Using a multivariate Cox proportional hazards model to adjust for age and left ventricular mass index the presence of nonsustained ventricular tachycardia was strongly and independently associated with increased all cause mortality and cardiovascular morbidity (relative risk 12.7, confidence interval 4.1-38.7; p is less than 0.0001). CONCLUSIONS: The presence of nonsustained ventricular tachycardia in hypertensive Black males with left ventricular hypertrophy is a strong and independent predictor of cardiovascular events. (c)2000 by Le Jacq Communications, Inc. PMID- 11416621 TI - A Missed Opportunity: Our Failure to Control Hypertension Optimally. AB - At a time when we know from randomized trials that control of hypertension will reduce morbidity and mortality from all of its complications, only 27% of the 50 million hypertensives in the U.S. have their blood pressures controlled to less than 140/90 mm Hg. Reasons for this failure to control blood pressure are many and include reluctance to use multidrug regimens, particularly in combination tablets, and failure to set and pursue blood pressure goals which should be discussed in advance with the patient by explaining the risks of elevated blood pressure and the proven benefits of optimal control. (c)2000 by Le Jacq Communications, Inc. PMID- 11416622 TI - An Update on the Diagnostic Evaluation of the Hypertensive Patient. AB - Hypertension is the most common reason for adult medical office visits in the U.S. today. With increasing longevity, an aging population, a greater prevalence of obesity, and a falling threshold for indications for high blood pressure treatment, the diagnostic evaluation of the hypertensive patient assumes a major role in office practice. The majority of patients who are being assessed for newly discovered hypertension will benefit from a relatively simple evaluation without the use of high cost procedures. Evaluation includes a careful history for both blood pressure related findings and other cardiovascular risk factors, a physical examination aimed at identifying possible specific etiologies and target organ damage, as well as some basic laboratory assays. In a rapid and relatively inexpensive fashion, the patient then can be directed toward nonpharmacologic treatment, drug therapy, or more specialized investigations as needed. (c)2000 by Le Jacq Communications, Inc. PMID- 11416623 TI - Hypertension as a Risk for Erectile Dysfunction: Implications for Sildenafil Use. AB - Hypertension is one of several risk factors for erectile dysfunction. Others include the cardiovascular risk factors of low levels of high density lipoprotein, high total cholesterol, smoking, and diabetes. Certain antihypertensive drugs, especially the thiazide diuretics, can contribute to erectile dysfunction. Sildenafil is the first oral agent available for the treatment of erectile dysfunction. It works in patients with psychogenic as well as organic erectile dysfunction, including men with hypertension and men on antihypertensive medicines. Sildenafil alone causes small drops in systolic and diastolic blood pressure. When administered to patients on antihypertensive drugs it was again associated with small drops in arterial pressure and no increase in adverse events compared to patients not taking antihypertensive drugs. When sildenafil is given in the setting of organic nitrates (nitric oxide donors), large falls in blood pressure may occur. Sildenafil is absolutely contraindicated in the setting of organic nitrate use. (c)2000 by Le Jacq Communications, Inc. PMID- 11416624 TI - Substance Abuse and Hypertension. AB - Substance abuse and hypertension are an important health concern, especially in adolescent and young adults presenting with elevated blood pressure and associated cardiovascular conditions. Illicit drugs including cocaine, marijuana, amphetamines, and methylenedioxymethamphetamine remain potential sources of acute or newly diagnosed hypertension. Cocaine is a powerful vasoconstrictor clearly related to hypertensive crises, myocardial infarction, arrhythmias, sudden death, strokes, and seizures, although not a definite cause of chronic hypertension. Potential therapies for cocaine induced hypertension and vasoconstriction include nitroglycerin, alpha-blockade combined alpha-A -blockade, and calcium channel blockers. The use of A -blockers alone should be avoided because of possible paradoxical rise in blood pressure including slight increases in systolic and diastolic blood pressure. Amphetamines and the newly abused 3,4 methylenedioxymethamphetamine are powerful sympathetic stimulants which may mimic the action of cocaine. Ecstasy, or 3,4-methylenedioxymethamphetamine, has additional severe psychiatric effects and should be considered in psychoses, panic disorders, and attempted suicide with new onset hypertension. The treatment approaches to 3,4-methylenedioxymethamphetamine toxicity needs further investigation. (c)2000 by Le Jacq Communications, Inc. PMID- 11416625 TI - Epidemiological and Clinical Aspects of Obesity Related Hypertension. AB - Substantial evidence from epidemiological data supports a link between obesity and hypertension. However, the relationship between the two disorders is not straightforward, and most likely represents the interaction of demographic, genetic, hormonal, renal, and hemodynamic factors. In the obese patient, hypertension can be reversed by weight loss. Furthermore, weight loss can prevent, or at least delay, the development of hypertension in patients with high normal blood pressure. Weight reduction should be the first line treatment in every obese hypertensive patient. However, patients will often need pharmacological intervention in conjunction with weight loss. (c)2000 by Le Jacq Communications, Inc. PMID- 11416626 TI - Current Concepts of Pharmacotherapy in Hypertension - Fixed Dose Combination Antihypertensive Therapy: Is This the Future? PMID- 11416627 TI - Common Questions and Answers in the Management of Hypertension - Everyday Practice in Hypertension: Herbal Remedies for High Blood Pressure. PMID- 11416629 TI - Clarify the Message! Improve Outcome in the Management of Hypertension. PMID- 11416628 TI - Analysis of Recent Papers in Hypertension. PMID- 11416630 TI - Comparison of Automated and Mercury Column Blood Pressure Measurements in Health Care Settings. AB - OBJECTIVES: Use of automated (electronic) blood pressure measurement devices to obtain clinic blood pressure measurements is becoming increasingly widespread in health care settings; their comparability with manual mercury sphygmomanometer readings is uncertain. Current guidelines for screening, diagnosis, and treatment of hypertension in clinical practice are based on clinical trials and epidemiological evidence derived from readings taken with auscultatory devices (usually mercury sphygmomanometers). This study was carried out to assess whether use of automated oscillometric devices in the clinical setting led to differences in classification of blood pressure levels at the 140 mm Hg systolic and 90 mm Hg diastolic thresholds compared to readings obtained with a standard mercury sphygmomanometer. DESIGN: We compared the blood pressure readings obtained with three automated devices that are widely available in the U.S. (one Dinamap Plus Model 8710 and two Dinamap Model 1846SX) and the readings taken by a single trained research assistant with a manual mercury column device in the emergency department and the outpatient medicine clinic of a large urban teaching hospital. The devices tested were those in normal use in the setting. The order in which the readings were taken was varied randomly. The sensitivity and specificity of the Dinamap readings compared to the mercury column device as the gold standard was calculated. RESULTS: The mean diastolic blood pressure in the three groups of patients studied was 7.3 mm Hg, 2.4 mm Hg, and 3.4 mm Hg lower with the Dinamap devices than the mercury column device (p is less than 0.001 for all comparisons). The mean systolic blood pressure readings were 1.0 mm Hg (p equals 0.06), 6.7 mm Hg (p is less than 0.001), and 4.2 mm Hg (p is less than 0.001) higher with the Dinamap device than the mercury column device. The difference between Dinamap and mercury column systolic blood pressure readings tended to increase at pressures greater than 140 mm Hg, whereas the diastolic blood pressure differences remained uniform throughout the blood pressure range. The sensitivity of the Dinamap readings compared to mercury column readings for classifying individuals as hypertensive was 73% for persons with elevated systolic blood pressure only, 51% for persons with both systolic and diastolic blood pressure elevation, and 10% for person with diastolic blood pressure elevation only. CONCLUSIONS: The Dinamap devices tested appeared to yield systematically biased blood pressure readings, which could alter the assessment of hypertension prevalence and control in clinical populations. Quality assurance and outcome researchers should attempt to document the type of device used to obtain blood pressure measurements noted in medical records, and be cognizant that small systematic errors in measurement could affect a setting's hypertension control performance. (c)2000 by Le Jacq Communications, Inc. PMID- 11416631 TI - Vasopeptidase Inhibition: A New Approach to the Management of Cardiovascular Disease. AB - Vasopeptidase inhibitors, a new class of cardiovascular compounds, inhibit both neutral endopeptidase, an enzyme that helps in the breakdown of vasodilator substances, and ACE. Simultaneous inhibition of neutral endopeptidase and ACE enhances peptides with vasodilatory properties, such as atrial natriuretic peptide, brain natriuretic peptide, and C type natriuretic peptide, and inhibits the production of the vasoconstrictor angiotensin II. The properties of these natriuretic peptides and their function in the regulation of the cardiovascular system are reviewed. Clinical results with vasopeptidase inhibitors and other therapeutic modalities based on the natriuretic peptide system in the treatment of hypertension and heart failure are also reviewed. Omapatrilat, an agent that has been recently evaluated, is an effective agent in lowering diastolic and particularly systolic blood pressures in a broad range of populations and may have beneficial effects beyond blood pressure control. The mechanism of action of omapatrilat and clinical results with this compound are discussed. (c)2000 by Le Jacq Communications, Inc. PMID- 11416632 TI - Combination Drug Therapy in the Management of Hypertension: When, With What, and How? PMID- 11416633 TI - The Role of Multiple Drug Therapy for Controlling Hypertension in African Americans. AB - Hypertension among black Americans explains much of the black health disadvantage. Compared with white Americans, blacks develop hypertension at a younger age and have higher rates of stage 3 hypertension (i.e., blood pressure is greater than 180/110 mm Hg). Black Americans tend to wait longer for treatment after diagnosis, causing a longer duration of elevated blood pressure and greater target organ damage. To arrive at the recommended target blood pressure of less than 130/85 mm Hg (in patients with diabetes or renal failure) or 125/75 mm Hg (in patients with proteinuria greater than 1 g/24 hours), all antihypertensive drug classes can be considered. Tight blood pressure control in black Americans frequently requires the use of more than just one agent (monotherapy). Low dose combination therapy, such as a calcium channel blocker and an ACE inhibitor, can solve many of the clinical issues that must be considered when treating hypertension in this patient group. (c)2000 by Le Jacq Communications, Inc. PMID- 11416634 TI - Preeclampsia: New Thoughts on an Ancient Problem. AB - Preeclampsia is a hypertensive disorder unique to pregnancy and is of unknown cause. It is both a maternal as well as fetal syndrome. It affects 2%-10% of nulliparas, and is more prevalent when preexisting hypertension, renal disease, or diabetes is present. The main clinical features of the disorder are maternal hypertension, proteinuria, and abnormal renal, hepatic, and central nervous system function. The fetal syndrome includes growth restriction, intrauterine death, and premature delivery. Abnormalities in the development of the placental circulation are believed to be important causal features of the pathogensis of preeclampsia and are present early in pregnancy. The link between the placental disease and the maternal syndrome remains mysterious. Clinical trials of low dose aspirin and calcium in the prevention of preeclampsia have not demonstrated significant benefits of treatment. The current approach to treatment is close monitoring of maternal and fetal health, bed rest, judicious use of antihypertensive therapy to prevent maternal complications, and seizure prophylaxis with magnesium sulfate. (c)2000 by Le Jacq Communications, Inc. PMID- 11416635 TI - Secondary Hypertension: Detection and Management for the Primary Care Provider. AB - Of hypertensive patients, 95% have primary (essential) hypertension. The remaining 5% of adults with hypertension have an identifiable or "secondary" disorder responsible for the elevated blood pressure. Although relatively rare, the diagnosis of secondary hypertension is important as these forms require specific treatments based on the underlying pathophysiology and are potentially curable. Secondary causes of hypertension include renal disease, such as chronic parenchymal disease and renovascular hypertension, a number of endocrinopathies, such as primary aldosteronism, pheochromocytoma, or thyroid disease, as well as a variety of miscellaneous causes including obstructive sleep apnea and substance abuse. An evaluation for secondary causes in all hypertensive patients is not necessary or cost effective. Clinical clues obtained from a careful history, physical examination, and laboratory assessment guide the clinician through an appropriate evaluation for identifiable causes. A combination of the clinician's index of suspicion and interpretation of data provide guidance in the choice of diagnostic tests and therapies to target the mechanisms contributing to poor blood pressure control. (c)2000 by Le Jacq Communications, Inc. PMID- 11416636 TI - 1999 Clinical Advisory Treatment of Hypertension and Diabetes. AB - The last Joint National Committee Report on the Detection, Evaluation, and Treatment of High Blood Pressure (JNC-VI) was published in 1997. Periodically, the National Heart, Lung, and Blood Institute has issued updates or advisories dealing with specific topics whereas featured recommendations may have changed as a result of new studies or new analyses. Recent and forthcoming updates include Hypertension in Pregnancy as well as Hypertension in Renal Disease. Presented below is an update (advisory) on the Treatment of Hypertension and Diabetes. (c)2000 by Le Jacq Communications, Inc. PMID- 11416637 TI - Can Diastolic Blood Pressure be Excessively Lowered in the Treatment of Isolated Systolic Hypertension? AB - In older populations, and those with prevalent cardiovascular disease, there is increasing evidence that reducing diastolic blood pressure below a level of 80-85 mm Hg is associated with increased risk of cardiovascular end points and all cause mortality. We sought to confirm this finding in participants in Systolic Hypertension in the Elderly Program, a placebo controlled trial of blood pressure reduction in older adults with isolated systolic hypertension. We found that, among subjects randomized to active treatment, for each 5 mm Hg lower achieved value in diastolic blood pressure increases the risk for stroke by 14%, coronary heart disease by 8%, and all cardiovascular disease 11%. This effect was not observed in the placebo group. Significant increases in cardiovascular disease risk were especially noted among patients for whom diastolic blood pressure was 60 mm Hg. This brief report suggests that older persons who develop large drops in diastolic blood pressure while undergoing treatment for isolated systolic hypertension may deserve special monitoring. (c)2000 by Le Jacq Communications, Inc. PMID- 11416638 TI - Current Concepts of Pharmacotherapy in Hypertension - Alpha-Adrenergic Blocking Drugs: Evolving Role in Clinical Medicine. PMID- 11416639 TI - Common Questions and Answers in the Management of Hypertension - Fish Oil for High Blood Pressure: Another Fish Story? PMID- 11416640 TI - Analysis of Recent Papers in Hypertension. PMID- 11416641 TI - Clinical Trials: A Commentary. PMID- 11416642 TI - Comforting News. PMID- 11416643 TI - Antihypertensive Drug Utilization in Hypertensive Veterans With Complex Medication Profiles. AB - A pharmacy prescription database was used to identify patients at high risk for drug-related problems. Of the 1054 patients in the study, 687 had a diagnosis of hypertension. The utilization of antihypertensive medications was captured at three periods over 24 months (12 months before enrollment, at enrollment, and 12 months after enrollment). The diagnosis of hypertension and coexisting diseases were identified at enrollment. There were 238 (34.6%) with diabetes, 333 (48.5%) with coronary artery disease, 64 (9.3%) with congestive heart failure, and 244 (35.5%) with none of these coexisting conditions. At Period 3, 44.7% of patients without coexisting diseases received calcium channel blockers, followed closely by diuretics (41.4%). Calcium channel blockers were used significantly more frequently than any other drug category for these patients (p less than 0.05). For patients with hypertension and diabetes, ACE inhibitors were used by 62%, and this was significantly more frequently than any other category (p less than 0.03). Diuretics (52.1%) were utilized significantly more frequently than calcium channel blockers (42.9%) (p less than 0.043). For patients with hypertension and congestive heart failure, diuretics were utilized significantly more than any other category (70.3%, p less than 0.03), and ACE inhibitors were utilized significantly more often than any other category except diuretics (68.8%, p less than 0.0001). This study examined antihypertensive utilization in specific patients (rather than as a function of total drugs), making the results different from those of previous reports. This study demonstrates better adherence to recommended guidelines than previous studies have suggested. While Beta blockers and diuretics were utilized frequently in these patients, statistics suggest that there is still room for improvement in the utilization of these important drugs. This paper describes the utilization of antihypertensive medications in nine Veterans Affairs Medical Centers. (c)2000 by Le Jacq Communications, Inc. PMID- 11416644 TI - The Effects of Candesartan Cilexetil in Isolated Systolic Hypertension: A Clinical Experience Trial. AB - An 8-week, multicenter, open-label, clinical experience trial evaluated the efficacy of candesartan cilexetil either alone (34%) or as add-on therapy (66%) in 1014 patients with untreated or uncontrolled isolated systolic hypertension defined as systolic blood pressure 140-179 mm Hg/diastolic less than 90 mm Hg. Candesartan cilexetil 16 mg once daily was given initially and was up-titrated to 32 mg once daily at week 2 or week 4 if systolic blood pressure remained at or above 140 mm Hg. Overall, candesartan cilexetil reduced both blood pressures by 16.5A+/-0.6/4.5A+/-0.2 mm Hg from 158A+/-0.4/81A+/-0.2 to 142A+/-0.6/76A+/-9.6 mm Hg with a control rate (systolic blood pressure less than 140 mm Hg) of 49%. Of the 492 (51%) patients remaining on the lower dose of candesartan cilexetil (40% as monotherapy; 60% as add-on), candesartan cilexetil 16 mg reduced blood pressure by 19.7A+/-1.0/5.5A+/-0.3 mm Hg and 62% of patients were controlled. For the 475 (49%) patients uncontrolled on candesartan cilexetil 16 mg and titrated to 32 mg (30% as monotherapy, 70% as add-on), the dose increase further reduced blood pressure by 8.9A+/-0.4/3.8A+/-0.1 mm Hg at week 8, and 36% of patients not responsive to the lower dose achieved blood pressure control on the higher dose. This dose response from candesartan cilexetil 16 mg to 32 mg was seen across age, sex, and race. Overall, tolerability was well maintained. Most adverse events were relatively infrequent, and only 8% withdrew due to adverse events; the most frequent adverse effects were dizziness (7%), headache (6%), and upper respiratory tract infection (5%). In conclusion, in this clinical experience trial for patients with isolated systolic hypertension, candesartan cilexetil 16 mg to 32 mg once daily produced a dose-related decrease in systolic blood pressure with a lesser decrease in diastolic blood pressure, resulting in a substantial decrease in pulse pressure. The dose response without dose dependent adverse effects was consistent across age, sex, and race, demonstrating that candesartan cilexetil is a therapeutic option for patients with isolated systolic hypertension. (c)2000 by Le Jacq Communications, Inc. PMID- 11416645 TI - Genes for Essential Hypertension: Hype, Help, or Hope? AB - Identification of genetic variation predisposing to high blood pressure is an active area of research. Characterization of genes for hypertension may permit focused efforts at primary prevention, may predict responses to pharmacologic interventions, and may identify individuals at risk for target organ damage. To date, genes for several rare types of hypertension have been identified, but such genes appear to play no important role in essential hypertension. Variants of the genes encoding the elements of the renin-angiotensin system have been extensively studied, with inconsistent results. It appears that such variants account for only a small part of blood pressure variation but may identify individuals at risk for blood pressure salt-sensitivity. Further successes in finding genes for hypertension will result from advances in genotyping, improved characterization of environmental factors impacting on genotype variation, and longitudinal studies of the evolution of pre-hypertension to hypertension. (c)2000 by Le Jacq Communications, Inc. PMID- 11416646 TI - The Eye as a Target Organ: An Updated Classification of Hypertensive Retinopathy. AB - The purpose of this paper is to recognize the eye as a hypertensive target organ that should be considered for risk stratification and subsequent therapeutic intervention. Revision of the traditional Keith, Wagner, and Barker classification of hypertensive retinopathy to a user-friendly vascular-based pathophysiologic classification should help. A simple, two-grade classification of hypertensive retinopathy that encourages the use of ophthalmoscopy to evaluate retinovascular changes is suggested. Utilization of this revised retinopathy classification will facilitate the evaluation of the hypertensive patient. (c)2000 by Le Jacq Communications, Inc. PMID- 11416647 TI - Management of Hypertension: Contributions of Clinical Trials. AB - The current state of hypertension management would not be possible without clinical trials, which have provided the most conclusive evidence of benefits from pharmacologic and lifestyle therapies. This review briefly covers the advancements in clinical trial methodology and in hypertension management. It focuses on three aspects of hypertension management, for which clinical trials gave conclusive findings where there were previously mixed findings from observational studies. The limitations of clinical trials are briefly discussed, as well. (c)2000 by Le Jacq Communications, Inc. PMID- 11416648 TI - Primary Prevention of Hypertension in Women. AB - Cardiovascular disease remains the leading cause of death in women in the U.S. In women greater than 45 years of age, 60% of Caucasian women and 79% of African American women have hypertension. It is estimated that 30% of elderly women (A365 years) are diagnosed with isolated systolic hypertension. Most risk reduction strategies for hypertension have traditionally focused on detection and treatment of the disease. Minor efforts have been aimed at primary prevention strategies for hypertension. A focus on lifestyle issues, which contribute to the development of hypertension in higher risk groups, may be beneficial, especially in premenopausal and perimenopausal women. These factors include potassium and sodium intake, obesity, physical inactivity, and excessive alcohol consumption. In this paper, the interventions for the primary prevention of hypertension based on these and other lifestyle factors will be reviewed, with special attention to the role they have in women. (c)2000 by Le Jacq Communications, Inc. PMID- 11416649 TI - Detection of Coronary Disease in Patients With Hypertension: A Resolved Issue? AB - When patients with hypertension experience chest pain, exercise ECG ischemia-like ST segment changes are frequent and often falsely indicate significant coronary artery disease; these patients often have normal coronary angiograms. Several alternative noninvasive tests have been proposed, and recently, the relative accuracy and reliability of the available techniques have been assessed. We have reviewed some of the relevant literature on the diagnostic tests employed in the clinical setting, including recent reports that stress echocardiography as more accurate than perfusion scintigraphy in identifying significant epicardial coronary artery disease in patients with hypertension. The low specificity of myocardial scintigraphy, which measures cardiac function and wall motion abnormalities, probably relates to the fact that this method traces perfusion abnormalities not necessarily caused by epicardial coronary artery disease, possibly due to microvascular disease and not necessarily causing obvious wall motion abnormalities. (c)2000 by Le Jacq Communications, Inc. PMID- 11416651 TI - Current Concepts of Pharmacotherapy in Hypertension - Doxazosin--What Does the Future Hold for it? PMID- 11416650 TI - Isolated Systolic Hypertension: An Update. PMID- 11416652 TI - Common Questions and Answers in the Management of Hypertension - Everyday Care of the Hypertensive Patient: Use of ACE Inhibitors in Fertile Women. PMID- 11416654 TI - A Year of Progress. PMID- 11416653 TI - Analysis of Recent Papers in Hypertension. PMID- 11416655 TI - The Prevention and Treatment of High Blood Pressure. PMID- 11416656 TI - The Relationship Between Body Weight and the Prevalence of Isolated Systolic Hypertension in Older Subjects. AB - The rise in systolic blood pressure and the increasing risk of isolated systolic hypertension with advancing age have been attributed, in part, to increased peripheral vascular resistance. Available evidence also suggests that at any given level of arterial pressure, overweight patients with hypertension have a higher cardiac output and a lower total peripheral vascular resistance than lean patients. The lower peripheral resistance associated with overweight patients may attenuate the rise in systolic blood pressure and modify the prevalence of isolated systolic hypertension with age. Using data from the third National Health and Nutrition Examination Survey we examined the relationship between body weight and the age-related rise in systolic blood pressure, and determined the implication of this relationship for the prevalence of isolated systolic hypertension in the older adult population. The age-related rise in systolic blood pressure and the prevalence of isolated systolic hypertension were higher among the lean than in the overweight (odds ratio 1.29 and p value 0.039). The prevalence of isolated systolic hypertension increased by 56% from 6.2% in the fourth to 9.7% in the first body mass index quartile. Among the overweight subjects the prevalence of isolated systolic hypertension continued to decrease until the body mass index exceeded 31.6. OBJECTIVE: To examine the relationship between body weight and the age-related rise in systolic blood pressure, and determine the implication of this relationship for the prevalence of isolated systolic hypertension in the older adult population. DESIGN: A cross-sectional study. SETTING: The study began with a 1-hour home visit for a household interview and ended a few weeks later with a 4-hour physical examination at a mobile examination center. PARTICIPANTS: A sample of the civilian, non institutionalized population of the U.S. MAIN OUTCOME MEASURES: Age-related rise in systolic blood pressure and the prevalence of isolated systolic hypertension. RESULTS: The age-related rise in systolic blood pressure and the prevalence of isolated systolic hypertension were higher among the lean than in the overweight. The prevalence of isolated systolic hypertension decreased with increased body mass index of 31.6. Systolic blood pressures increased in obese than nonobese subjects. CONCLUSIONS: Body mass index less than 25 is associated with an increase in the age-related rise in systolic blood pressure and the prevalence of isolated systolic hypertension in the older population. (c)2000 by Le Jacq Communications, Inc. PMID- 11416657 TI - Angiotensin II Antagonists in the Treatment of Hypertension: More Similarities Than Differences. AB - As the class of angiotensin-II antagonists expands, it becomes relevant to know if there are differences in antihypertensive efficacy among the various agents. Prior to regulatory approval, all agents have been evaluated vs. placebo. We excerpted the placebo-corrected reductions in diastolic blood pressure for angiotensin II antagonist monotherapy, using objective regulatory review data from the U.S. product circulars. Both systolic and diastolic blood pressure reductions were very similar. In 25 randomized clinical trials that compared angiotensin II antagonists to other classes, equivalent antihypertensive efficacy was demonstrated at recommended doses. Data pooled from 51 clinical trials showed comparable weighted average diastolic blood pressure reductions (not placebo corrected) for monotherapy with losartan, valsartan, irbesartan, candesartan, and telmisartan. Reductions in systolic blood pressure paralleled the changes in diastolic blood pressure. Somewhat smaller responses were observed with eprosartan, although this was based on fewer patients. Thus, there appears to be little clinically significant difference in blood pressure efficacy among the six marketed angiotensin II antagonists in the treatment of hypertension. (c)2000 by Le Jacq Communications, Inc. PMID- 11416658 TI - Why Objective Monitoring of Compliance is Important in the Management of Hypertension. AB - Hypertensive patients often experience poor adherence to treatment, a frequent cause of uncontrolled blood pressure. In this study, we have evaluated whether or not the use of electronic monitoring for drug adherence is a useful approach to identify and correct compliance problems in hypertensive patients, which may ultimately enhance the effect of antihypertensive therapy. Sixty-nine treated patients with an office blood pressure greater than 140/90 mm Hg were enrolled in this study. With patient consent, current antihypertensive therapy was dispensed in electronic pillboxes that record the time and date of each opening without changing the drug regimen. The intention was to provide physicians with objective measurements of drug compliance. The monitoring of compliance per se without any other intervention induced a marked decrease of blood pressure in the whole group (from 159/104A+/-23/12 mm Hg to 143/92A+/-20/15, meansA+/-standard deviation, p less than 0.001). A complete normalization of blood pressure (less than 140/90 mm Hg) was obtained in one third of the patients (group 1, n=23) during the monitoring period. A significant improvement of blood pressure control was found in another third (group 2, n=23), whereas in the remaining patients (group 3, n=23) no change in blood pressure was observed. The distribution of individual compliance values, as well as the mean compliances was comparable in the three subgroups. Conversely, the compliance reports have identified several potentially overtreated patients in group 1, a large number of patients with a poor adherence to the prescribed therapy in all groups, and patients who clearly needed a change in pharmacotherapy mainly in group 3. Thus, our results suggest that electronic monitoring of compliance can considerably enhance the efficacy of antihypertensive therapy in patients with uncontrolled hypertension. This procedure should be used more extensively in clinical practice whenever the blood pressure response to therapy appears insufficient. (c)2000 by Le Jacq Communications, Inc. PMID- 11416659 TI - Blood Pressure Therapy in the Elderly: An Observation in Octogenarians. AB - The incidence of hypertension increases with age. Although there is ample evidence regarding the benefit of treating elderly hypertensive patients, resulting in decreases in cardiovascular morbidity and mortality, the adverse effects of treatment are also frequently reported in these often frail individuals. Therefore, it is difficult to determine the level to which blood pressure should be decreased and what type of antihypertensive medication is most effective and best tolerated. The approach to treating hypertension in octogenarians at the Cleveland Clinic Florida was reviewed. An analysis of 100 consecutive charts of hypertensive octogenarians treated by internists from January 1, 1993, with a minimum of 1 year follow up, was carried out. The mean age of the population studied was 84.7 years (range, 80-97A+/-4.13 years). The mean initial blood pressure was 152/83 mm Hg, and the mean final blood pressure was 145/76 mm Hg (pless than 0.01). The initial number of medications used was 1.2, which rose to 1.6 by the final reading (pless than 0.0001). The most frequently used drug classes were calcium channel blockers, followed by diuretics and ACE inhibitors. Most of the octogenarians tolerated their antihypertensive medication well. (c)2000 by Le Jacq Communications, Inc. PMID- 11416660 TI - Diuretics and Circadian Rhythm of Blood Pressure. AB - In patients with essential hypertension it has been postulated that the lack of nocturnal fall in blood pressure (nondippers) is associated with more serious end organ damage than in patients whose blood pressure falls during the night (dippers). Although the mechanism of abnormal circadian blood pressure rhythm remains unclear, our recent studies have revealed the importance of kidneys in the genesis of the rhythm. We found that in patients who have a sodium sensitive type of hypertension, blood pressure failed to fall during the night, and both sodium restriction and diuretics shifted circadian rhythm of blood pressure from nondipper to dipper. In addition, we identified the sodium sensitivity of blood pressure as an independent risk factor for cardiovascular morbidity in patients with essential hypertension. Diuretic-based treatment may have an additional therapeutic advantage to relieve the renal and cardiovascular risks by different ways: systemic blood pressure reduction and normalization of circadian blood pressure rhythm. (c)2000 by Le Jacq Communications, Inc. PMID- 11416661 TI - Current Concepts of Pharmacotherapy in Hypertension - Chronotherapeutics and Its Role in the Treatment of Hypertension and Cardiovascular Disease. PMID- 11416662 TI - Effects of Stress and Behavioral Interventions in Hypertension - Panic Attacks, Hyperventilation, and Hypertension. PMID- 11416663 TI - Common Questions and Answers in the Management of Hypertension - Home Blood Pressures Kits. PMID- 11416664 TI - Analysis of Recent Papers in Hypertension. PMID- 11416665 TI - Treating Hypertension: Calcium Channel Blockers, Diuretics, Beta Blockers, ACE Inhibitors: Is There a Difference? PMID- 11416666 TI - Isolated Systolic Hypertension Is a Strong Predictor of Cardiovascular and All Cause Mortality in the Middle-Aged Population: Warsaw Pol-MONICA Follow up Project. AB - OBJECTIVE: To assess the long-term risk of cardiovascular disease and all-cause mortality associated with various categories of hypertension, particularly isolated systolic hypertension. MATERIALS AND METHODS: Three independent random samples of the general Warsaw population aged 35-64 years (2490 men and 2405 women screened in 1984, 1988, and 1993) were followed for (1) cardiovascular death until 1996 and (2) all-cause death until 1998. Methods of the baseline surveys were consistent with the manual of the WHO MONICA Project. Isolated systolic hypertension was defined as systolic blood pressure greater than or equal to 140 mm Hg and diastolic blood pressure less than 90 mm Hg and was classified into two stages: stage I, when 90 is less than systolic blood pressure is less than 160 mm Hg; and stage II, for subjects with systolic blood pressure greater than or equal to 160 mm Hg. The Cox proportional hazard model was used to calculate the multivariable-adjusted relative risk of cardiovascular disease and all-cause mortality attributed to the particular hypertension category. RESULTS: Compared to normotension, stage I was associated with a 53%-60% increase in age standardized cardiovascular mortality rates in both genders and with a 70% increase in all-cause mortality in men. A significant and independent association (after multivariable adjustment) was found only for all-cause mortality in men. Stage II was an independent predictor of cardiovascular mortality in men and in women, and of all-cause mortality in men. In both genders, the risks of examined end points associated with stage II were comparable to the risks associated with isolated systolic hypertension. Isolated diastolic hypertension did not significantly increase the risk of all-cause mortality in either gender or of cardiovascular mortality in men. In women the risk of cardiovascular disease associated with isolated diastolic hypertension was almost threefold higher compared with the normotensive group, but the risk was not statistically significant. CONCLUSION: In the middle-aged Polish population, isolated systolic hypertension is a much better predictor of cardiovascular disease and all-cause mortality than isolated diastolic hypertension. The association was stronger for stage II than for stage I. The risk of examined end points associated with stage II were similar to those associated with combined systolic-diastolic hypertension. (c)2000 by Le Jacq Communications, Inc. PMID- 11416667 TI - Target Organ Damage in Hypertension. AB - Millions of people have hypertension, and a lesser percentage have both diabetes and hypertension. Hypertension and diabetes together are the most common etiologies for development of renal failure and death from cardiovascular causes in the Western world. The number of people who achieve a goal blood pressure shown to markedly reduce cardiovascular risk is 27.4%. Moreover, only 11% of the population in the NHANES III database achieved the recommended goal of less than 130/85 mm Hg for those with diabetes. Consequently, the rate of people starting dialysis continues to climb and while cardiovascular-associated mortality has been reduced, morbidity has not changed appreciably. The presence of target organ injury, left ventricular hypertrophy, and microalbuminuria signifies a long duration of pressure overload. All of these manifestations of pressure overload are associated with higher cardiovascular event rates and renal disease progression when compared to people without these manifestations. This difference in event rates may be related to shorter duration and/or intensity of blood pressure elevation or achievement of blood pressure goals in people with established hypertension. It is clear that patients with evidence of target organ injury need aggressive blood pressure lowering to the goals set forth by the JNC VI in order to both regress injury to the cardiovascular and renal systems and help maximally reduce cardiovascular and renal injury. (c)2000 by Le Jacq Communications, Inc. PMID- 11416668 TI - Nonsteroidal Anti-Inflammatory Drugs and Hypertension. AB - Nonsteroidal anti-inflammatory drugs are among the most widely prescribed medications. Their effect on blood pressure has been monitored, and many small studies have determined a potential relationship between their use and elevation of blood pressure. These drugs may affect blood pressure by inhibiting prostaglandin synthesis, which may affect arteriolar smooth muscle tone and natriuresis. Since many patients with conditions such as osteoarthritis require treatment and also have hypertension, even modest elevations in blood pressure or inhibition of antihypertensive medication efficacy resulting from non steroidal anti-inflammatory drugs can be of significant clinical and public health importance. This review finds that certain drugs (e.g., indomethicin, piroxicam, and naproxen) may cause clinically relevant elevations in blood pressure in hypertensive patients. Aspirin and sulindac do not appear to elevate blood pressure significantly, even in hypertensive patients. Ibuprofen and other nonsteroidal anti-inflammatory drugs appear to have an intermediate blood pressure effect. Cyclo-oxygenase-2 inhibitors such as refecoxib and celecoxib have been shown to cause mild elevations in blood pressure, but further studies are needed to evaluate the full magnitude and population distribution of this effect. (c)2000 by Le Jacq Communications, Inc. PMID- 11416669 TI - Physician Role in Lack of Awareness and Control of Hypertension. AB - OBJECTIVES: To describe the systolic and diastolic blood pressure criteria used by community physicians to: 1) establish a diagnosis of hypertension; and 2) increase dosage or change drug therapy in treated hypertensives. METHODS: A multiethnic population sample of adults at or above 40 years old was surveyed, by random digit phone dialing in a major metropolitan area, regarding blood pressure measurement and hypertension awareness and treatment status. Respondents with and without known hypertension were asked to: 1) identify their health care providers; and 2) agree to sign and return a release form allowing the investigators to obtain copies of their medical records. Medical records received were abstracted in a standardized format. RESULTS: Medical records were obtained for 169 patients who collectively had 940 visits with 175 different providers during a consecutive 2-year period. Overall, 25% of these patients had 24-month average blood pressures of at or above 140/90 mm Hg without a diagnosis of hypertension being recorded by a physician. Over two-thirds of the undiagnosed had systolic blood pressure of 140-159 mm Hg, with diastolic pressure of less than 90 mm Hg. In persons on antihypertensive drugs (n equals 81), the average blood pressure was 147/86 mm Hg, and only 24% met the Joint National Committee-VI blood pressure goal of less than 140/90 mm Hg. A visit-level analysis indicated that when diastolic blood pressure was greater than 90 mm Hg, physicians intensified drug therapy 24% of the time, but intensification actions occurred in only 4% of visits when systolic pressure was less than 140 mm Hg and diastolic pressure was less than 90 mm Hg. Kaplan-Meier survival curves and multiple logistic regression modeling demonstrated that almost no action would be taken for persistently elevated systolic blood pressure over 20 consecutive visits. CONCLUSIONS: Community physicians do not give equal weight to systolic blood pressure of greater than 140 mm Hg as to diastolic pressure of greater than 90 mm Hg in diagnosing hypertension and intensifying treatment. This is a major determinant of the current level of unawareness of hypertension and uncontrolled, known hypertension found in national surveys. (c)2000 by Le Jacq Communications, Inc. PMID- 11416670 TI - Inhibiting Both RAS and SNS for High Blood Pressure Control: Myths and Facts. PMID- 11416671 TI - Current Concepts of Pharmacotherapy in Hypertension - Early Identification of Those at High Risk for Hypertension and Rationale for Initial Therapy. PMID- 11416672 TI - Effects of Stress and Behavioral Interventions in hypertension - Headache and Hypertension: Something Old, Something New. PMID- 11416673 TI - Common Questions and Answers in the Management of Hypertension - Is Diastolic Pressure Passe? PMID- 11416674 TI - Analysis of Recent Papers in Hypertension. PMID- 11416675 TI - Ambulatory blood pressure monitoring: how important is it in estimating risk or guiding therapy? PMID- 11416676 TI - The hypertensive heart: new observations and evolving therapeutic imperatives. PMID- 11416677 TI - Antihypertensive efficacy of candesartan in comparison to losartan: the CLAIM study. AB - An 8-week, multicenter, double-blind, randomized, parallel-group, forced titration study was conducted to evaluate the antihypertensive efficacy of candesartan vs. losartan in 654 hypertensive patients with a diastolic blood pressure between 95 and 114 mm Hg from 72 sites throughout the U.S. Eligible patients were randomized to candesartan cilexetil 16 mg once daily, or losartan 50 mg once daily. Two weeks following randomization, patients doubled the respective doses of their angiotensin receptor blockers for an additional 6 weeks. At week 8, candesartan cilexetil lowered trough systolic/diastolic blood pressure by a significantly greater amount than did losartan (13.3/10.9 mm Hg with candesartan cilexetil vs. 9.8/8.7 mm Hg with losartan; p less than 0.001). At the same period, candesartan cilexetil also lowered peak blood pressure by a significantly greater amount than did losartan (15.2 to 11.6 mm Hg with candesartan cilexetil vs. 12.6 to 10.1 mm Hg with losartan; p less than 0.05). There were statistically significantly (p less than 0.05) higher proportions of responders and controlled patients in the candesartan cilexetil group (62.4% and 56.0%, respectively) than in the losartan group (54.0% and 46.9%, respectively). Both treatment regimens were well tolerated; 1.8% in the candesartan cilexetil group and 1.6% in the losartan group withdrew because of adverse events. In conclusion, this forced-titration study confirms that candesartan cilexetil is more effective than losartan in lowering blood pressure when both are administered once daily at maximum doses. Both drugs were well tolerated. (c)2001 by Le Jacq. PMID- 11416678 TI - Diastolic dysfunction in arterial hypertension. AB - Left ventricular diastolic properties are important markers of pump function and are frequently abnormal when myocardial insults alter tissue structure. Alterations can be limited to the early diastolic phase (early active relaxation) or to late diastolic filling (late ventricular compliance), but more often involve regulation of both phases of diastole. In asymptomatic patients with arterial hypertension, left ventricular relaxation is often prolonged, independently, at least in part, of cardiac loading conditions and left ventricular geometry, but this abnormality is associated with early signs of systolic dysfunction. Uncontrolled hypertension, diabetes, and obesity are most often associated with ischemic heart disease and impaired diastolic function. Reducing blood pressure with antihypertension therapy will reduce myocardial afterload, regress LVH, and improve systolic and diastolic function. In patients with symptoms of CHF with a normal ejection fraction, however, changes in therapy may be indicated. Greater emphasis should be placed on using medications that decrease myocardial load, but also reduce the effects of neurohormonal activation. (c)2001 by Le Jacq Communications, Inc. PMID- 11416679 TI - Low birth weight contributes to the excess prevalence of end-stage renal disease in African Americans. AB - The risk of hypertension and related target organ damage is much greater in African Americans than in Caucasians. The risk of hypertensive end-stage renal disease is approximately five-fold higher in African Americans. Many studies have shown that low birth weight is strongly associated with increased risk of hypertension, stroke, and myocardial infarction. However, until recently the relationship between birth weight and hypertension-related diseases was not clearly established in African Americans. Moreover, it was also unclear if low birth weight in humans heightened the risk for end-stage renal disease. This is a critical gap in the literature, since low birth weight occurs at twice the rate in African Americans as among Caucasians. We identified a significant relationship between end-stage renal disease and low birth weight in both African Americans and Caucasians. Given the higher rates of low birth weight in African Americans, differences in fetal development may, therefore, contribute to the racial disparity in end-stage renal disease. Continued study of the biological factors linking early development with later risk of hypertension-related diseases is important and may shed light on racial disparities in health outcomes. (c)2001 by Le Jacq Communications, Inc. PMID- 11416680 TI - Less salt intake or more salt excretion: is hypertension preventable? AB - Hypertension is preventable by modification of the common source (i.e., sodium chloride) and by prevention of weight gain and increased consumption of "good" nutrients. Unfortunately, the necessary societal changes are unlikely to occur. Individual behavioral approaches are weak. A pharmacologic approach as a "crutch" for our unsuccessful control of a "common source" epidemic may work and have a major impact on morbidity and mortality due to hypertensive diseases. Once-a-day diuretics may be a better health dose than once-a-day vitamins for our nutrient rich society. (c)2001 by Le Jacq Communications, Inc. PMID- 11416681 TI - Choosing initial antihypertensive drug therapy for the uncomplicated hypertensive patient. AB - Choosing the initial antihypertensive drug for the uncomplicated hypertensive patient is an important and frequent event for the primary care physician. Patients' first experience with antihypertensive drug therapy will likely affect their long-term perception of hypertension treatment. The choice should be made on the basis of sound scientific data and from the patient's perspective and needs. The drug should be taken once a day, should have proven efficacy in hypertension control and cardiovascular morbidity and mortality reduction, and should have as few side effects as possible. Low-dose thiazide diuretics meet this description, although the need to monitor electrolytes may make them less than ideal. The angiotensin II receptor antagonist class, with side-effects similar to those of placebo in controlled trials, is the most attractive from the patient's perspective, although outcome trial data do not yet exist proving that hypertension treatment with angiotensin II receptor antagonists reduces cardiovascular events. The angiotensin-converting enzyme inhibitors or angiotensin II receptor antagonists, with their low side-effect profiles and unique effects on vascular remodeling, are attractive second choices to combine with a diuretic if needed, although low-dose diuretic/Beta blocker combinations have also been shown to lower blood pressure with minimal side effects. At present, ensuring adequate long-term hypertension control is the most important aspect of hypertensive care, and which antihypertensive drug(s) the physician chooses can greatly affect the hypertensive patient's ability to achieve and to maintain long-term blood pressure control. (c)2001 by Le Jacq Communications, Inc. PMID- 11416682 TI - Clinical pharmacology of the angiotensin receptor antagonists. PMID- 11416683 TI - Why is hypertension more common in African Americans? PMID- 11416684 TI - Renal artery stenosis in hypertension: what about the "other" alternative? PMID- 11416687 TI - Poor adherence to hypertension therapy: whose responsibility is it? PMID- 11416688 TI - Management of hypertension in pregnancy. PMID- 11416689 TI - Working group report on high blood pressure in pregnancy. AB - This report updates the 1990 National High Blood Pressure Education Program Working Group Report on High Blood Pressure in Pregnancy and focuses on classification, pathophysiology, and management of the hypertensive disorders of pregnancy. Using evidence-based medicine and consensus, this report updates contemporary approaches to hypertension control during pregnancy by expanding on recommendations made in the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC VI). The recommendations to use K5 for determining diastolic pressure and to eliminate edema as a criterion for diagnosing pre-eclampsia are discussed. In addition, the use of blood pressure increases of 30 mm Hg systolic or 15 mm Hg diastolic as a diagnostic criterion has not been recommended, as available evidence shows that women in this group are not likely to suffer increased adverse outcomes. Management considerations are made between chronic hypertension that is present before pregnancy and those occurring as part of the pregnancy-specific condition preeclampsia, as well as management considerations in women with comorbid conditions. A discussion of the pharmacologic treatment of hypertension in pregnancy includes recommendations for specific agents. The use of low-dose aspirin, calcium, or other dietary supplements in the prevention of pre-eclampsia is described, and expanded sections on counseling women for future pregnancies and recommendations for future research are included. Once again we thank Dr. Ray Gifford, Jr., and his committee for volunteering their time to produce this important report. We hope it helps the busy clinician prevent and manage a very important problem. PMID- 11416690 TI - Prognostic value of direct, continuous ambulatory blood pressure monitoring in essential hypertension. AB - This study assessed the prognostic value of ambulatory vs. clinic blood pressure measurement in 688 hypertensives who had undergone pretreatment 24-hour intra arterial ambulatory blood pressure monitoring. A total of 157 first events were recorded over a follow-up period of 9.2A+/-4.4 years. Ambulatory systolic or diastolic blood pressure parameters (whether 24-hour mean, daytime mean, or nighttime mean) or ambulatory pulse pressure provided independent prognostic information in conjunction with clinical variables. The most predictive models contained the ambulatory systolic blood pressure parameters. Age, male gender, South Asian origin, diabetes mellitus, and previous cardiovascular disease were additional independent predictors of events. In a subgroup of 295 uncomplicated patients, 24-hour ambulatory pulse pressure was an independent predictor of left ventricular mass index and maximal carotid intima-media thickness. Baseline clinic blood pressure parameters did not provide independent information for the prediction of events or target organ damage. Therefore, in this study, ambulatory blood pressure proved to be superior to clinic measurement for cardiovascular risk stratification. However, the routine use of ambulatory blood pressure monitoring is not currently recommended, mainly because of a lack of outcome trials based on the treatment of ambulatory blood pressure levels. (c)2001 by Le Jacq Communications, Inc. PMID- 11416691 TI - Microalbuminuria: what is it? Why is it important? What should be done about it? AB - Microalbuminuria (MA) is defined as persistent elevation of albumin in the urine, of 30-300 mg/day (20-200 microg/min). These values are less than the values detected by routine urine dipstick testing, which does not become positive until protein excretion exceeds 300-500 mg/day. Use of the albumin-to-creatinine ratio is recommended as the preferred screening strategy for all diabetic patients. MA is measured in spot morning urine obtained from the patient in the office and sent for measurement of both albumin and creatinine. A value above 0.03 mg/mg suggests that albumin excretion is above 30 mg/day and therefore MA is present. MA should be checked annually in everyone, and every 6 months within the first year of treatment to assess the impact in patients started on antihypertensive therapy. MA is an established risk factor for renal disease progression in type 1 diabetes and its presence is the earliest clinical sign of diabetic nephropathy. In addition, a number of studies suggest that MA is an important risk factor for cardiovascular disease and defines a group at high risk for early cardiovascular mortality in both type 2 diabetes and essential hypertension. MA also signifies abnormal vascular permeability and the presence of atherosclerosis. Among nondiabetic patients with essential hypertension, MA is associated with higher blood pressures, increased serum total cholesterol, and reduced serum high density lipoprotein cholesterol. Thus, taken together these data support the concept that the presence of MA is the kidney's notice to the physician/patient that there is a problem with the vasculature. MA can be reduced, and progression to overt proteinuria prevented, by aggressive blood pressure reduction. The National Kidney Foundation recommends that blood pressure levels be maintained at or below 130/80 mm Hg in anyone with diabetes or renal disease. This should be accomplished with antihypertensive agents that prevent the rise in MA and hence prevent development of proteinuria. Such agents are angiotensin-converting enzyme inhibitors, angiotensin receptor blockers and, to a lesser extent, Beta blockers, non-dihydropyridine calcium antagonists, and diuretics. In summary, the presence of MA is a marker of endothelial dysfunction and a harbinger of markedly enhanced cardiovascular risk. All patients with diabetes and/or hypertension should be screened for the presence of microalbuminuria with use of spot morning urine. To maximize prevention of MA development, the following goals should be instituted: 1) blood pressure should be maintained at less than 130/80 mm Hg and a low-salt, moderate-potassium diet instituted; 2) in diabetics, HbA1c should be kept at less than 7%; 3) in obese patients, a weight loss program should be implemented, with a goal BMI of less than 30; and 4) the physician and patient, working together, should maintain low-density lipoprotein cholesterol at less than 120 mg/dL, and less than 100 mg/dL if diabetes is present. (c)2001 by Le Jacq Communications, Inc. PMID- 11416692 TI - Treating hypertension in the elderly--whom to treat, when, and with what? PMID- 11416693 TI - Direct vasodilators and their role in hypertension management: minoxidil. AB - Minoxidil is a direct vasodilator that has been in use for over two decades. It is used primarily to reduce blood pressure in hypertensives who have been poorly controlled on various multidrug regimens. Although minoxidil is extremely effective, its usefulness is limited by its tendency to increase the pulse rate and to trigger salt and water retention. The latter may be incapacitating in some patients. Therefore, minoxidil is typically administered with both a diuretic and an agent that can control the pulse rate, such as a beta blocker. Minoxidil has several other side effects that may limit its use, including hypertrichosis, aggravation of myocardial ischemia and/or left ventricular hypertrophy, and (infrequently) pericardial effusions. If a patient's hypertensive pattern is sufficiently severe to warrant contemplation of minoxidil therapy, referring the patient to a hypertension specialist should be strongly considered. (c) 2001 by LeJacq Communications, Inc. PMID- 11416694 TI - Behavioral treatment of hypertension. PMID- 11416695 TI - How often should "surveillance EKGs" be obtained in the follow-up of asymptomatic hypertensives? PMID- 11416698 TI - Diuretics revisited--again. PMID- 11416699 TI - Gender differences in vascular compliance in young, healthy subjects assessed by pulse contour analysis. AB - OBJECTIVES: Sex hormones exert important effects on the vasculature. Female sex hormones have been reported to enhance endothelial function, reduce oxidative stress, and protect against atherosclerosis. However, the effects of estrogen on vascular compliance have not been studied. Recently, noninvasive instrumentation that estimates vascular compliance by recording the radial artery pulse contour has been introduced. Reductions in the oscillatory or reflected component of the diastolic waveform have been observed in various clinical conditions, including hypertension, diabetes mellitus, and congestive heart failure, and may reflect endothelial dysfunction at the site of resistance vessels. In this study the authors examined gender-related vascular compliance in a cohort of young, healthy, predominantly nonsmoking, medication-free men and women to determine the influence of cardiovascular risk factors, including family and social history, serum lipids, plasma homocysteine, and insulin levels on vascular compliance. METHODS: The volunteers, consisting of 151 healthy men and women (mean age 24A+/ 4 years) completed a questionnaire detailing family and social history, medication use, and exercise habits. Large (C1) and small (C2) vessel compliance and various cardiovascular parameters were derived from arterial pulse wave contour analysis. Systolic, diastolic, and mean arterial blood pressure, pulse pressure, and pulse rate were determined simultaneously by oscillometry. Blood for fasting serum lipids, plasma homocysteine, and serum insulin were obtained in a subset of 135 subjects. RESULTS: The questionnaire revealed that 38% of parents had a history of hypertension, 31% had dyslipidemia, and 15% had coronary heart disease. C2 was lower in subjects with parental dyslipidemia. Compared to men, women had lower C2; lower systolic blood pressure, mean arterial pressure, and pulse pressure; higher serum high-density lipoprotein cholesterol; lower serum triglycerides; and lower plasma homocysteine, but similar serum insulin levels. C1 correlated with height and pulse pressure, whereas C2 was proportional to height and weight and inversely related to systemic vascular resistance. Multivariate regression analysis showed that stroke volume, total vascular impedance, cardiac output, female gender, and systemic vascular resistance independently predicted changes in C2, but that height was not a significant factor. CONCLUSIONS: Women have reduced C2 despite lower systolic blood pressure and pulse pressure and more favorable lipid and homocysteine levels. C2 is independent of height and is lower in subjects with parental dyslipidemia. These data indicate that female sex hormones have unexpected negative effects on small vessel compliance. They may help to explain why premenopausal women hospitalized for myocardial infarction have higher mortality rates than men of the same age. PMID- 11416700 TI - Treatment of hypertension to prevent stroke: translating evidence into clinical practice. AB - Although programs such as the National High Blood Pressure Education Project emphasize that all patients with hypertension should be treated, it is perhaps natural for physicians to question the usefulness of aggressively treating those patients who have modestly elevated blood pressure levels but are otherwise healthy, asymptomatic individuals. Using the example of stroke, this review summarizes the clinical trial data demonstrating the efficacy of antihypertensive therapy in patients with severe and nonsevere forms of hypertension. Suggestions are made that will help practitioners to apply results from the published literature in clinical practice. The evidence suggests that treating patients with nonsevere hypertension would prevent a larger proportion of the population wide burden of stroke than treating only those with more severe hypertension. Helping physicians to understand and apply this evidence will bring us closer to the goal of population-wide treatment and control of high blood pressure. PMID- 11416701 TI - Emergency room management of hypertensive urgencies and emergencies. AB - Hypertensive crisis affects upward of 500,000 Americans each year. Although the incidence of hypertensive crisis is low, affecting fewer than 1% of hypertensive adults, more than 50 million adult Americans suffer from hypertension. Presentation of a patient with severe hypertension to the emergency room demands immediate evaluation, prompt recognition of a hypertensive emergency or urgency, and the prompt institution of appropriate therapeutic measures to prevent progression of target-organ damage and to avoid a catastrophic event. Hypertensive emergencies are severe elevations in blood pressure that are complicated by evidence of progressive target-organ dysfunction such as coronary ischemia, disordered cerebral function, a cerebrovascular event, pulmonary edema, or renal failure. Although therapy with parenteral antihypertensive agents may be initiated in the emergency department, these patients warrant prompt admission to an intensive care unit where continuous monitoring of blood pressure can be assured during therapy. PMID- 11416702 TI - Alcohol consumption and hypertension. AB - Many observational studies have shown a relationship between three or more alcoholic drinks daily and hypertension. Reduction in alcohol intake is associated with lowering of blood pressure in randomized clinical trials: each drink per day reduction in intake lowers systolic and diastolic blood pressure by approximately 1 mm Hg. Although regular alcohol consumption seems to reduce the incidence of atherothrombotic cardiovascular events, excessive alcohol intake increases the risk of many medical and psychosocial problems. For persons with hypertension who drink excessively, average maximum alcohol intake of one drink per day in women and two drinks per day in men is a reasonable goal, if drinking is not otherwise contraindicated. PMID- 11416703 TI - The role of home blood pressure monitoring in hypertension control. AB - Hypertension is estimated to affect as many as 50 million adults in the United States. Despite nationally developed, evidence-based guidelines for care and the availability of numerous effective medications for the treatment of hypertension, control rates for this condition remain very low. Only 21%-27% of diagnosed patients in the United States are controlled to blood pressure levels of less than 140/90 mm Hg.1 Factors contributing to the poor hypertension control rate include adherence issues, variation in provider practice patterns, and the lack of a systematic approach to tracking and maintaining blood pressure control. Home monitoring combined with systematic treatment and follow-up by skilled health care professionals can have a positive impact in all three of these areas. PMID- 11416704 TI - Current concepts of pharmacotherapy in hypertension--ophthalmically administered beta blockers and their cardiopulmonary effects. AB - Early clinical studies revealed that timolol and other topical beta blockers were effective in reducing intra-ocular pressure, without the side effects associated with other antiglaucoma agents. However, because persons with cardiovascular or respiratory diseases were generally excluded from many of these early studies, the risk of serious cardiovascular and respiratory side effects was seriously underestimated. Once these drugs were made available to the general population, reports of systemic side effects began to proliferate. Very quickly, adverse effects from topical beta blockade became "old news." Despite this recognition, many treating physicians remained unaware of the potential for systemic beta blockade from topically applied beta blockers. A significant portion of a topically administered dose of a beta blocker can be absorbed and thereby affect systemic beta blockade. Sensitivity to systemic beta blockade can be quite dramatic in certain highly susceptible patients, particularly those with either cardiac or pulmonary abnormalities. Careful review of patients' medications will generally lessen, but not completely eliminate, the risk of undesired complications attributable to topical beta blockade. PMID- 11416705 TI - Why are we doing so badly with the control of hypertension? Poor compliance is only part of the story. PMID- 11416706 TI - What's the scoop on salt? PMID- 11416708 TI - Primary aldosteronism: a practical approach to diagnosis and treatment. AB - Primary aldosteronism (PA) may account for as many as 10%-14% of hypertension cases. The plasma aldosterone concentration/plasma renin activity ratio is a simple screening test for PA that should be performed in all patients with refractory/severe hypertension, spontaneous or provoked (by diuretics) hypokalemia, or a requirement for excessive potassium supplementation to maintain normokalemia. PA can be confirmed by a fludrocortisone suppression test or 24 hour urine collection for aldosterone. Confirmatory testing should be followed by high-resolution computerized tomography of the adrenal glands to distinguish bilateral hyperplasia or an adenoma. A solitary tumor greater than 1 cm in size in a younger patient is an indication for surgery; all other (nondiagnostic) findings should be followed by bilateral adrenal venous sampling (if available) to identify a unilateral cause of PA. Treatment for a lateralizing positive study is surgical; spironolactone or another mineralocorticoid receptor antagonist is the treatment of choice for a nonlateralizing study. If adrenal venous sampling is not readily available, patients may be successfully treated pharmacologically. PMID- 11416710 TI - High exposure to nevirapine in plasma is associated with an improved virological response in HIV-1-infected individuals. AB - OBJECTIVE: To explore relationships between exposure to nevirapine and the virological response in HIV-1-infected individuals participating in the INCAS trial. METHODS: The elimination rate constant of plasma HIV-1 RNA (k) was calculated during the first 2 weeks of treatment with nevirapine, zidovudine and didanosine in 51 antiretroviral-naive HIV-1-infected patients. The relationships between the value of k, the time to reach an undetectable HIV-1 RNA concentration in plasma (< 20 copies/ml) and the success of therapy after 52 weeks of treatment as dependent variables and the exposure to nevirapine, baseline HIV-1 RNA and baseline CD4 cell count as independent variables, were explored using linear regression analyses, proportional hazard models and logistic analyses, respectively. RESULTS: The value of k for HIV-1 RNA in plasma was positively and significantly associated with the mean plasma nevirapine concentration during the first 2 weeks of therapy (P = 0.011) and the baseline HIV-1 RNA (P = 0.008). Patients with a higher exposure to nevirapine reached undetectable levels of HIV 1 RNA in plasma more rapidly (P = 0.03). From 12 weeks on, the median nevirapine plasma concentration was significantly correlated with success of therapy after 52 weeks (P < 0.02). CONCLUSIONS: A high exposure to nevirapine (in a twice daily regimen) is significantly associated with improved virological response in the short as well as in the long term. These findings suggest that optimization of nevirapine concentration might be used as a tool to improve virological outcome in (antiretroviral-naive) patients treated with nevirapine. PMID- 11416711 TI - T cell expansions in lymph nodes and peripheral blood in HIV-1-infected individuals: effect of antiretroviral therapy. AB - OBJECTIVE: To evaluate dynamics in CD8 T cell expansions during highly active antiretroviral therapy (HAART). DESIGN: Various T cell subsets were isolated from blood and lymph nodes and analysed for T cell receptor (TCR) diversity. METHODS: TCR complementarity determining region 3 (CDR3) spectratyping and single-strand conformation polymorphism (SSCP) analyses were performed in combination with sequencing to assess clonality of the subsets. RESULTS: Strongly skewed CDR3 patterns in total CD8 cells and the CD8 subsets CD45RO+CD27+ and CD45RO-CD27+ showed substantial dynamics in dominant CDR3 sizes, resulting in relative improvement of CDR3 size diversity in the first months of therapy. During sustained treatment, TCR diversity changed only moderately. SSCP profiles confirmed oligoclonality of TCR CDR3 perturbations. Various dominant CDR3 sizes for CD4 and CD8 T cells present in lymph nodes, but not in peripheral blood mononuclear cells, before the start of therapy emerged in peripheral blood early during therapy. CONCLUSIONS: HAART induces substantial changes in CD8 TCR diversity, eventually resulting in improvement of the repertoire. Clonal expansions observed in lymph nodes before therapy were observed in peripheral blood after therapy, suggesting that recirculation of CD4 and CD8 T cells from lymph nodes contributes to the early T cell repopulation. Decreased immune activation and possibly naive T cell regeneration subsequently decreased clonal expansions and perturbations in the CD8 TCR repertoire. PMID- 11416712 TI - Soluble CD23 in cerebrospinal fluid: a marker of AIDS-related non-Hodgkin's lymphoma in the brain. AB - BACKGROUND: AIDS-related non-Hodgkin's lymphoma (NHL) includes systemic lymphomas, often with brain involvement, and primary central nervous system (CNS) lymphomas. OBJECTIVE: To examine if measurement of soluble CD23 (sCD23) in cerebrospinal fluid (CSF) is useful in the diagnosis and follow-up of AIDS related NHL. METHOD: sCD23 was measured by enzyme-linked immunosorbent assay and EBV DNA by nested polymerase chain reaction for a group of 134 patients. The NHL group included 14 patients with primary HIV-1 CNS lymphoma, 12 patients with brain involvement of systemic HIV-1 NHL and 10 patients with systemic HIV-1 NHL without brain involvement. These were compared with HIV-1-infected patients with cerebral toxoplasmosis (19), progressive multifocal leukoencephalitis (PML; 8) and AIDS-related dementia (17) and with asymptomatic HIV-1 carriers (54) and uninfected individuals (50). The levels of sCD23 were compared with the presence of Epstein-Barr virus (EBV) DNA in CSF. RESULTS: Significantly higher levels of sCD23 were found in the CSF of the patients with brain lymphoma than in those with systemic NHL (P < 0.002) or with cerebral toxoplasmosis, PML and AIDS related dementia (P < 0.0001). The sensitivity and specificity of sCD23 in CSF as a marker for detection of brain NHL were 77% and 94%, respectively. High levels of sCD23 were found in CSF from patients with brain NHL independently of the presence (18 out of 26) or absence (8 out of 26) of EBV DNA. CONCLUSIONS: The sCD23 in CSF of HIV-1-infected patients may represent an additional, non-invasive marker for diagnosis of brain involvement in AIDS-related NHL. PMID- 11416713 TI - Differences in HIV disease progression by injection drug use and by sex in the era of highly active antiretroviral therapy. AB - BACKGROUND: The advent of highly active antiretroviral therapy (HAART) has reduced the incidence of most AIDS-related opportunistic illnesses (OI) and death in HIV-infected individuals. We investigated whether there are demographic disparities in HIV disease progression in the HAART era compared with before. METHODS: HIV-infected patients in an urban HIV clinical practice in the USA were compared using survival methods for time to a new AIDS-defining OI or death in therapeutic era 1 (monotherapy and combination therapy; 1990--1995; n = 2016) versus era 2 (HAART; 1996--1999; n = 2165). RESULTS: A total of 1037 (51.4%) events occurred in era 1; 666 (30.8%) events occurred in era 2. In women, the median disease-free survival time increased by 14% (CD4 cell counts > 200 cells/mm(3) at baseline) and 34% (CD4 cell counts < or = 200) in era 2 compared with era 1, whereas for men it increased by 43 and 100%. The relative hazard (RH) of progression for women compared with men in era 2 compared with era 1 was 1.34. For injecting drug use (IDU), disease-free survival time increased by 16% and 34% in era 2 compared with era 1, whereas non-IDU improved by 65 and 135%. The RH of progression for IDU compared with non-IDU in era 2 compared with era 1 was 1.39. No significant differences were detected by race or other HIV transmission risk group. CONCLUSION: Disease-free survival time was extended with the use of HAART, but these gains were not equally distributed by sex and IDU in our cohort. PMID- 11416714 TI - Phenotypic hypersusceptibility to non-nucleoside reverse transcriptase inhibitors in treatment-experienced HIV-infected patients: impact on virological response to efavirenz-based therapy. AB - BACKGROUND: Enhanced susceptibility to non-nucleoside reverse transcriptase inhibitors (NNRTI) was recently described in association with increased resistance to nucleoside analogs (nucleoside reverse transcriptase inhibitors; NRTI). OBJECTIVES: To determine the prevalence of NNRTI hypersusceptibility, the genotypic correlates, and its impact on virologic response to efavirenz-based salvage therapy. METHODS: Genotype and phenotype testing was performed retrospectively on baseline isolates from 30 patients who received salvage therapy containing efavirenz. NNRTI hypersusceptibility was defined as a 50% inhibitory concentration (IC(50)) of < 0.5 that of the wild-type control. RESULTS: Eight isolates had major NNRTI mutations. Among the 22 isolates with no major NNRTI mutations, 11 (50%) were hypersusceptible to efavirenz, 10 (45%) to delavirdine, and eight (36%) to nevirapine. Among eight isolates with NNRTI mutations, NNRTI resistance was present, but at lower than expected levels. The number of NRTI mutations was correlated inversely with the fold decrease in susceptibility to efavirenz (Spearman's rho, -0.57; P = 0.005), delavirdine (rho, -0.43; P = 0.04), and nevirapine (rho, -0.69; P < 0.001). Excluding subjects with NNRTI mutations, subjects with efavirenz hypersusceptibility at baseline had significantly better virologic suppression over 24 weeks than those without efavirenz hypersusceptibility (P < 0.001). CONCLUSION: NNRTI hypersusceptibility is common in heavily treated but NNRTI naive patients and is related directly to NRTI resistance mutations. Among patients receiving efavirenz-containing regimens, NNRTI hypersusceptibility was associated with an improved virologic outcome after 24 weeks of therapy. A reversal of phenotypic resistance was seen in patients with NNRTI mutations in the presence of multiple NRTI mutations, but no obvious virologic benefit of this phenomenon was seen in this study. PMID- 11416715 TI - Comparison of the effectiveness of non-nucleoside reverse transcriptase inhibitor containing and protease inhibitor-containing regimens using observational databases. AB - OBJECTIVES: To compare the effectiveness of first protease inhibitor (PI) containing and non-nucleoside reverse transcriptase inhibitor (NNRTI)-containing regimens. METHODS: Data were analysed from three large HIV patient databases: Apache HIV Insight (APACHE), Target Management Services (TMS) and Clinical Partners (CP). The effectiveness of therapy was the time taken for HIV-1 RNA to fall below detectable levels on first highly active antiretroviral therapy regimen (PI- or NNRTI-containing) and the subsequent time to failure (two consecutive detectable measurements). Comparisons were made using proportional hazards models, adjusting for differences in age, sex, previous reverse transcriptase inhibitor use, calendar year and baseline viral load and CD4 T-cell count. RESULTS: The type of regimen was not associated with time to undetectable viral load in any of the three databases, all of which had high power to detect a difference. PI-containing regimens were significantly less likely to fail after reaching undetectable viral loads for APACHE and CP patients (relative hazard, 1.7; 95% confidence interval, 1.3--2.1 and relative hazard, 1.6; 95% confidence interval, 1.0--2.5 respectively). These results remained significant after allowing for an unmeasured confounder with moderate effect on risk. No significant association between time to failure and regimen was found for TMS patients, possibly due to low power (67% to detect a relative hazard of 1.5). No difference was found between regimens in the time taken for an increase of > 100 x 10(9)cells/l in CD4 T-cell count. In the APACHE database, those on NNRTI containing regimens were more likely to have a failing CD4 T-cell response. CONCLUSIONS: PI-containing regimens have a lower risk of treatment failure than NNRTI-containing regimens. PMID- 11416716 TI - Initiating co-trimoxazole prophylaxis in HIV-infected patients in Africa: an evaluation of the provisional WHO/UNAIDS recommendations. AB - OBJECTIVE: To evaluate the proposed WHO/UNAIDS criteria for initiating co trimoxazole prophylaxis in adult HIV-infected patients in Africa [WHO clinical stages 2--4 or CD4 count < 500 x 10(6) /l or total lymphocyte count (TLC) equivalent]. DESIGN: Observational cohort study of 5-year follow-up. SETTING: Adult HIV clinics, University of Cape Town, South Africa. METHODS: Effect of prophylactic low dose co-trimoxazole (480 mg per day or 960 mg three times per week) on survival and morbidity was assessed in patients stratified by WHO clinical stage, CD4 T-lymphocyte count or TLC. Patients receiving antiretroviral therapy were excluded. RESULTS: Co-trimoxazole reduced mortality [adjusted hazard ratio (AHR), 0.56; 95% confidence interval (CI), 0.33--0.85; P > 0.001] and the incidence of severe HIV-related illnesses (AHR, 0.52; 95% CI, 0.38--0.68; P < 0.001) in patients with evidence of advanced immune suppression on clinical (WHO stages 3 and 4) or laboratory assessment (TLC < 1250 x 10(6)/l or CD4 count < 200 x 10(6)/l). No significant evidence of efficacy was found in patients with WHO stage 2 or CD4 count 200--500 x 10(6)/l/TLC 1250--2000 x 10(6)/l. If we had applied the WHO/UNAIDS recommendations 88.3% of our patients would have received co-trimoxazole prophylaxis at their initial clinic visit. CONCLUSION: Co trimoxazole in HIV-infected adults from an area in which Pneumocystis carinii pneumonia is uncommon demonstrated a survival benefit consistent with previous randomized trials. Further studies are needed to assess the optimal time of commencement of prophylaxis, as widespread co-trimoxazole use will lead to increasing antimicrobial resistance to other major pathogens in Africa. PMID- 11416717 TI - Risk factors for and trends in gonorrhea incidence among persons infected with HIV in the United States. AB - OBJECTIVE: To determine the risk factors for and trends in gonorrhea infections among HIV-infected persons. DESIGN: Longitudinal review of medical records of HIV infected patients. METHODS: We analyzed data about HIV-infected patients obtained from 1991 to 1998 in over 100 facilities participating in the Adult/Adolescent Spectrum of HIV Disease Project. RESULTS: The overall incidence of gonorrhea was 9.5 cases per 1000 person--years. Factors associated with higher gonorrhea incidence (P < 0.01) included younger age, male--male sex, black race, HIV infection without AIDS (namely AIDS-defining opportunistic illness or CD4 cell count < 200 x 10(6) cells/l), and recent recreational use of injection or non injection drugs. There was an increase in the trend among men who have sex with men (P < 0.01) and a decrease in the trend among patients with heterosexual contact as their HIV exposure risk (P < 0.01). Among injection drug users there was no significant trend from 1991 to 1996, but there was an increase in gonorrhea incidence from 6.6 cases/1000 person-years in 1997 to 16.3 cases/1000 person--years in 1998. CONCLUSIONS: Following HIV diagnosis, some individuals continue to practice risky sexual behaviors which result in gonorrhea and may transmit HIV. The increase in the trend in gonorrhea incidence among HIV-infected men who have sex with men is of particular concern because it suggests an increase in risky sexual behaviors. These findings indicate a need for effective HIV prevention strategies that involve reducing risky sexual behaviors in HIV infected persons. PMID- 11416718 TI - Predictors of intrauterine and intrapartum transmission of HIV-1 among Tanzanian women. AB - OBJECTIVE: To examine predictors of vertical transmission of HIV-1 in Dar-es Salaam, Tanzania. DESIGN: Observational design. METHODS: Consenting HIV-1 infected pregnant women (n = 1078) were enrolled in a trial to examine the role of vitamin supplements. Intrauterine HIV-1 infection (HIV-positive at birth); intrapartum and early breastfeeding transmission (HIV-positive at 6 weeks among those uninfected at birth) were defined using the PCR. RESULTS: Of 734 infants who had a specimen taken at birth, 62 were HIV positive [8.4%; 95% confidence interval (CI),6.4--10.5%], whereas 59 infants were positive among 367 infants who were uninfected at birth and were retested at 6 weeks (16.1%; 95%CI, 12.3- 19.8%). In multivariate analyses, maternal CD4 cell count, viral load, and clinical stage were significant predictors of both definitions of transmission. Viral load of 50 000 copies/ml or more at delivery was associated with a 4.21 fold increase in risk of intrapartum and early breastfeeding transmission (95%CI, 1.59--11.13;P = 0.004). Babies who were HIV negative at birth and born before 34 weeks of gestation were 2.19 times more likely to become infected during intrapartum and early breastfeeding periods compared with those born after 37 weeks (95%CI, 1.19--4.04; P = 0.01). Gonorrhea at baseline was related to intrauterine transmission [multivariate risk ratio (RR), 5.50; 95%CI, 2.04- 14.81; P < 0.001] but not intrapartum and early breastfeeding transmission. Signs of lower genital infections at or after enrollment were also associated with transmission. CONCLUSIONS: Reducing prematurity, rate of HIV disease progression, and maternal viral load at or after delivery could help to reduce vertical transmission. Treatment of sexually transmitted infections at onset of prenatal care, about 20 weeks on average, was inadequate for prevention of transmission. Whether sustained clearance of lower genital tract infections result in reduced transmission remains to be determined. PMID- 11416719 TI - Prevalence and distribution of HHV-8 in different subpopulations, with and without HIV infection, in Spain. AB - OBJECTIVE: To estimate the seroprevalence of HHV-8 in several Spanish subpopulations with different risk levels of acquiring HIV-1 infection and from different geographical regions. DESIGN: Cross-sectional seroprevalence study. METHODS: A total of 1699 serum samples from blood donors (613), children under the age of 12 years (100), injecting drug users (IDU) (382), heterosexuals attending a sexually transmitted disease (STD) clinic (273) and homosexual men attending a STD clinic or a HIV-based hospital unit (331) were analysed for anti HHV-8 antibodies. The presence of antibodies against HHV-8 was tested with an indirect immunofluorescence assay (IFA). A subsample of HHV-8-positive samples was also tested for antibody titre against HHV-8. RESULTS: The overall seroprevalence of antibodies against HHV-8 for the blood donor population was 6.5% (7.0% in Andalusia, 8.0% in Catalonia and 4.5% in the Basque Country). None of the children tested positive for HHV-8. The HHV-8 prevalence was 86.7% in HIV positive homosexual men and 28.0% in HIV-negative homosexual men (P < 0.001). Of heterosexual men attending STD clinics, 17.2% tested positive for HHV-8; 11.5% of IDU tested positive for HHV-8. HHV-8 antibody titres by groups parallel the distribution of HHV-8 prevalence. No association between HHV-8 antibody titres and CD4 cell count or HIV viral load was identified. CONCLUSIONS: The HHV-8 prevalence among blood donors in Spain is higher than in Northern Europe and the USA, but is similar to that in Northern Italy. The distribution of HHV-8 is compatible with a sexually transmitted agent. The distribution of HHV-8 correlates with that of Kaposi's sarcoma but factors other than HHV-8 seem to explain the Kaposi sarcoma distribution. PMID- 11416720 TI - Transient or occult HIV-1 infection in high-risk adults. PMID- 11416721 TI - Changes in apoptosis after interruption of potent antiretroviral therapy in patients with maximal HIV-1-RNA suppression. PMID- 11416722 TI - Non-adherence to highly active antiretroviral therapy predicts progression to AIDS. PMID- 11416723 TI - Accumulation of lopinavir resistance-associated mutations over 3 years follow-up of patients on highly active antiretroviral therapy: implication in salvage therapy. PMID- 11416724 TI - Declining incidence of visceral leishmaniasis in HIV-infected individuals in the era of highly active antiretroviral therapy. PMID- 11416725 TI - Treatment of tuberculosis in HIV-infected patients: safety and antiretroviral efficacy of the concomitant use of ritonavir and rifampin. PMID- 11416726 TI - HIV infection may adversely affect clinical response to chloroquine therapy for uncomplicated malaria in children. PMID- 11416727 TI - Circumcision and male-to-male sexual transmission of HIV. PMID- 11416728 TI - Secondary structure predictions of HIV-1 reverse transcriptase provide new insights into the development of drug-resistance genotypes. PMID- 11416729 TI - Efavirenz plasma concentrations and efficiency. PMID- 11416731 TI - L-Carnitine for the treatment of highly active antiretroviral therapy-related hypertriglyceridemia in HIV-infected adults. PMID- 11416732 TI - Mother to child transmission: European Collaborative Study. PMID- 11416733 TI - BK virus encephalitis in a patient with AIDS and lymphoma. PMID- 11416734 TI - HIV dynamics and T-cell immunity after three structured treatment interruptions in chronic HIV-1 infection. AB - OBJECTIVE: To evaluate whether controlled re-exposures to autologous HIV-1 could boost HIV-specific immunity and limit virus replication in patients with chronic HIV-1 infection. PATIENTS AND DESIGN: Subjects with at least 2 years virus suppression during antiretroviral therapy and a CD4 : CD8 ratio > 1 were randomly assigned to interrupt highly active antiretroviral treatment (HAART) three times (n = 12) or to continue their previous HAART (n = 14). RESULTS: In 10/12 interrupter patients a rebound of HIV-1 RNA was detected in all three structured treatment interruptions (STI). Plasma virus doubling time was shorter during the first STI than in the second and third STI, corresponding to an average 13% reduction in viral basic reproductive rate. However, the mean time before plasma viral load rose to > 50 copies/ml was significantly shorter in the second and third STI. The average frequency of HIV-specific CD8 T cells in the interrupter patients at the end of the third STI cycle was significantly higher compared with the baseline and the end of the first STI. A substantial increase in HIV-specific CD8 T cell frequencies was found in four interrupter patients, whereas there were no changes in all 14 non-interrupter individuals. A weak p24-specific T helper response developed in 5/12 interrupter patients compared with no response in non interruptors, but these responses were transient and disappeared rapidly. CONCLUSION: The increase in the control of viral replication, and positive effects of STI on immune responses in this population should encourage the further development of HIV-specific immune-based therapeutic strategies. PMID- 11416735 TI - The virological and immunological consequences of structured treatment interruptions in chronic HIV-1 infection. AB - BACKGROUND: Some individuals with chronic HIV-1 infection have discontinued their drug therapy with consequent plasma virus rebound. In a small number of patients, a delayed or absent rebound in plasma virus load has been noted after drug cessation, apparently associated with prior drug interruptions and autologous boosting of HIV-1 specific immune responses. We hypothesized that cyclic structured treatment interruptions structured treatment interruptions (STI) could augment HIV-1 specific immune responses in chronic HIV-1 infection, which might help to control HIV-1 replication off therapy. METHODS: We initiated an STI pilot study in 10 antiretroviral treatment-naive HIV-1 chronically infected subjects with baseline CD4 T-cell counts > 500 x 10(6) cells/l and plasma viral load > 5000 copies/ml who received highly active antiretroviral therapy (HAART) for 1 year with good response (plasma viral load < 20 copies/ml for at least 32 weeks). Three cycles of HAART interruption were performed. RESULTS: In all of the patients viral load rebounded, but doubling times increased significantly between the first and third stops (P = 0.008), and by the third stop, six out of nine subjects had a virological set-point after a median 12 months off therapy that was lower than baseline before starting HAART (ranging from 0.6 log(10) to 1.3 log(10) lower than baseline) and in four it remained stable below 5000 copies/ml. Those subjects who controlled viral replication developed significantly stronger HIV-1 specific cellular immune responses than subjects lacking spontaneous decline (P < 0.05). During viral rebounds no genotypic or phenotypic changes conferring resistance to reverse trancriptase inhibitors or protease inhibitors was detected, but mean absolute CD4 T-cell counts declined significantly, although never below 450 x 10(6)/l and the mean value at 12 months off therapy was significantly higher than the pre-treatment level (P = 0.004). CONCLUSIONS: Our findings suggest that STI in chronic HIV-1 infection might augment HIV-1 specific cellular immune responses associated with a spontaneous and sustained drop in plasma viral load in some subjects but at the potential cost of lower CD4 T-cell counts. PMID- 11416736 TI - Detection of multiple accessory spleens in a patient with chronic idiopathic thrombocytopenia purpura. AB - Chronic idiopathic thrombocytopenia purpura is an immunologically mediated disorder resulting in disordered platelet kinetics and potentially life threatening thrombocytopenia. Failure of medical therapy is an indication for splenectomy, and responses are seen in 70% of patients after this procedure. An important cause of relapse after splenectomy is the presence of accessory spleens. The authors describe a patient with chronic idiopathic thrombocytopenia purpura who had a relapse of symptoms 4.5 years after splenectomy. Tc-99m heat denatured RBC planar and SPECT studies revealed the presence of multiple (more than 13) accessory spleens, but few of them were localized anatomically using computed tomography. PMID- 11416737 TI - Ga-67 and Tl-201 scintigraphy in extramedullary plasmacytoma: a case report. AB - Extramedullary plasmacytoma is a rare form of plasma cell tumor occurring in a wide variety of organs and tissues. Most tumors occur in the head and neck, involving the nasal cavity, paranasal sinuses, and upper airway. Tl-201 and Ga-67 scan findings in a patient with extramedullary plasmacytoma in the right side of the chest wall are presented. Only a few publications have described Tl-201 or Ga 67 uptake in extramedullary plasmacytoma. This is the first report of both Tl-201 and Ga-67 uptake in an extramedullary (or intramedullary) plasmacytoma. PMID- 11416738 TI - FDG positron emission tomographic, radioiodine, and MIBI imaging in a patient with poorly differentiated insular thyroid carcinoma. AB - Poorly differentiated insular thyroid carcinoma is now classified as a separate entity among other tumors of the thyroid gland. Its histologic pattern and its clinical course are regarded as intermediate between well differentiated and anaplastic thyroid cancer. Insular carcinoma accumulates I-131, but no data exist regarding its fluorodeoxyglucose (FDG) positron emission tomographic (PET) uptake. The authors report F-18 FDG PET, Tc-99m MIBI, and radioiodine imaging features in a 63-year-old patient with metastatic insular thyroid carcinoma. After total thyroidectomy (for poorly differentiated insular carcinoma pT3a), the patient was referred for radioiodine ablation. No signs of recurrence were present until 16 months later, when thyroglobulin levels increased. An I-131 scan showed a single lesion in the right lung, and further radioiodine treatment was administered (cumulative dose [530 mCi], 19,610 MBq I-131). Three years after the initial diagnosis, FDG-PET and Tc-99m MIBI scans were performed within 5 days during thyroxine treatment. After that, thyroxine substitution was withdrawn; 6 weeks later, an I-131 whole-body scan was performed. Both radioiodine and MIBI images showed increased tracer uptake in the known lung lesion. However, FDG PET showed a normal tracer distribution. Magnetic resonance and computed tomographic imaging confirmed a 12-mm lesion in the right upper lobe. These findings support the concept of the "flip-flop phenomenon" in insular thyroid carcinoma, an alternating pattern of metastases with either I-131 or FDG-uptake. Despite poorly differentiated histologic findings, glucose metabolism was not increased in this patient with an insular tumor. PMID- 11416739 TI - Evaluation of lymphatic drainage of cold thyroid nodules with intratumoral injection of Tc-99m nanocolloid. AB - PURPOSE: The lymphatic system may show variations throughout the entire body. Knowledge of the variations and aberrant lymph drainage are important when planning surgical treatment and radiotherapy. The aim of this study was to evaluate the intratumoral injection technique in the detection of lymphatic drainage of proved or possibly malignant cold thyroid nodules. MATERIALS AND METHODS: The study group consisted of 13 patients with palpable solitary cold thyroid nodules. None of the patients had cystic nodules on ultrasound examination. After fine-needle aspiration biopsy (at least 3 days later), 15 MBq (0.4 mCi) Tc-99m nanocolloid particles in a small volume (0.2 ml) were injected into the nodule. Dynamic images (60 frame x 1 minute) were acquired during the first hour, followed by static anterior and lateral images at 90 and 120 minutes. RESULTS: Radiopharmaceutical was present in the systemic circulation in two patients, possibly as a result of paratumoral injection. Eleven patients had intratumoral accumulation in early frames. Lymph nodes draining the thyroid nodule were visualized in 10 of 11 patients. In the dynamic acquisition period of 7 of 10 patients, the first draining lymph node could also be identified. Only 5 of 13 patients had malignant nodules. Lymphatic metastases were detected in one patient's specimens. CONCLUSIONS: Preoperative visualization of lymphatic pathways may be used when planning for thyroid cancer surgery. Sentinel node detection could yield valuable additional information in these patients. PMID- 11416740 TI - Severe hypertension in children with renovascular disease. AB - Renovascular disease is an important cause of hypertension in children and is associated with considerable morbidity and mortality risks. Secondary hypertension is more common in children than in adults, with children accounting for 75% to 80% of cases. In 70% of secondary hypertension in children, the cause is fibromuscular hyperplasia. Other associated conditions are aorto-aortitis, the midaortic syndrome, and Williams-Bueren syndrome. Imaging techniques have an important role in the early discovery of renal artery stenosis. Although renal arteriography remains the definitive method, noninvasive and less invasive radiographic procedures such as ultrasonography with duplex Doppler scanning and radionuclide scintigraphy have been used as adjunct diagnostic tools in children. The authors describe three young children with renovascular hypertension in whom dynamic radionuclide scintigraphy with Tc-99m MAG3 played an essential role in the diagnosis of renovascular hypertension. PMID- 11416741 TI - Incidental recognition of left subclavian vein obstruction on renal scintigraphy. AB - In a renal transplant recipient with persistently poor graft function, the flow phase of a renal scan incidentally revealed multiple venous collaterals with focally increased vascular activity near the left lobe of the liver (quadrate lobe). This was initially assumed to represent superior vena cava (SVC) obstruction. A renal biopsy was contemplated to exclude acute rejection because of a nondiagnostic flow phase (loss of a bolus effect). However, because the possibility of venous obstruction at the level of the subclavian and/or brachiocephalic veins (without involving the SVC) also existed, another renal scan was performed, with injection of radiotracer into the contralateral arm. This showed a patent SVC and reasonably preserved renal perfusion consistent with acute tubular necrosis. Subsequently, left subclavian vein obstruction was identified. The graft function improved with conservative management for acute tubular necrosis. These findings illustrate the danger of considering only SVC obstruction when collateral flow patterns and focal hot spots in the liver are present. PMID- 11416742 TI - F-18 FDG uptake in a bone infarct: a case report. AB - PURPOSE: The authors describe a patient with F-18 fluorodeoxyglucose (FDG) uptake resulting from a bone infarct of a rib. MATERIALS AND METHODS: A 52-year-old woman with a history of left breast cancer and total mastectomy 5 years earlier was examined for pain in her left rib cage in the past 2 weeks. A bone scan showed diffusely increased uptake in the left fifth rib, possibly suggestive of a metastasis. An FDG positron emission tomographic (PET) scan was performed to rule out metastatic disease. RESULTS: The FDG PET images showed two focal areas of increased uptake in the fifth and sixth left ribs, consistent with bone metastases. The patient underwent excision of a long segment of her left fifth rib and the pathological findings were consistent with bone infarct, without definite signs of metastatic disease. CONCLUSION: Bone infarct can take up FDG, probably as a result of an inflammatory process. PMID- 11416743 TI - Increased accumulation of Tl-201 in monostotic Paget's disease of the patella: evaluation with quantitative analysis. AB - Monostotic Paget's disease of the patella was detected with Tc-99m MDP and Tl-201 scans. Diffuse intense uptake of MDP in the left patella was observed on the blood-pool and late phases of the bone scan. Tl-201 imaging was performed to differentiate a malignant process and showed diffuse marked accumulation at the same site. Semiquantitative analysis of the patella region on both Tc-99m MDP and Tl-201 scans did not support a diagnosis of cancer. Radiographs showed the features of Paget's disease of the bone. Findings of a pathologic evaluation were compatible with the diagnosis of osteitis deformans. This case represents the unusual skeletal involvement of monostotic Paget's disease of the bone in the patella. Tl-201 accumulation in the Paget's lesion was suggested to be caused by increased metabolic activity of the lesion but was not indicative of a malignant process. PMID- 11416744 TI - The "SAPHO" syndrome: a case report of a patient with unusual bone scan findings. AB - PURPOSE: The authors describe the clinical and bone scintigraphic findings of the SAPHO syndrome, which is characterized by synovitis, acne, palmoplantar pustulosis, hyperostosis, and osteitis. The case report illustrates the significance of bone scintigraphy in the diagnosis. It shows that Tc-99m MDP scanning can detect signs of arthritis not seen with other imaging methods, because the arthritis is inflammatory in nature and does not always cause bone erosion. Thus it is not visualized on plain radiographs. Knowledge of this disorder may help in the differential diagnosis of arthritis. MATERIALS AND METHODS: Tc-99m MDP bone scintigraphy was used to diagnose arthritic changes. Whole-body and multiple delayed spot images were obtained in a 39-year-old diabetic, hypertensive woman who had tenderness in the plantar aspect of her heels. Swelling of the small and large joints of the feet, ankles, knees, hips, right sacroiliac joints, and shoulders was noted. She also had hydradenitis suppurativa and a history of a previous episode in which the arthritis improved after surgical treatment for the hydradenitis. RESULTS: All active joint lesions were visualized on the bone scan, including the arthritis, which was not detected with other imaging methods. They were all well demonstrated in the bone scintiscan. The scan findings, along with the presence of hydradenitis, led to the correct diagnosis of SAPHO syndrome. CONCLUSION: Tc-99m MDP bone scanning may be helpful in diagnosing arthritis as associated with the SAPHO syndrome. PMID- 11416745 TI - The volume effect of the stomach on intestinal activity on same-day exercise- rest Tc-99m tetrofosmin myocardial imaging. AB - PURPOSE: The aim of this study was to investigate quantitatively the volume effect of the stomach on the prevention of intestinal artifacts in the rest phase on same-day exercise--rest Tc-99m tetrofosmin myocardial perfusion scintigraphy. MATERIALS AND METHODS: The study was performed in 81 consecutive patients (43 men, 38 women; mean age, 57 +/- 9 years) who were referred for routine Tc-99m tetrofosmin exercise--rest SPECT myocardial perfusion imaging with normal scintigrams or completely normal inferior wall perfusion on visual assessment of SPECT images. Group A (control group, n = 41) and group B (n = 40) were given 200 ml milk to enhance the hepatobiliary clearance of tetrofosmin 30 minutes before the exercise SPECT acquisition study and just after the injection for the rest SPECT acquisition study. For the rest SPECT acquisition study, patients in group B were given, in addition to milk, a standard solid and fluid meal to enhance the volume of the stomach after the injection of Tc-99m tetrofosmin. In all patients, both exercise and rest SPECT acquisitions were started 45 minutes after the injection of Tc-99m tetrofosmin. Myocardial and abdominal activity were assessed on three separate views in the rest phase of the study, and the mean myocardium to-abdomen count ratios were calculated. In addition, the presence of interfering intestinal activity was assessed visually on SPECT images. Infracardiac activity was described as absent when it was equal to lung background. RESULTS: The myocardial-to-abdominal ratios were 1.38 +/- 0.05 and 1.95 +/- 0.04 in groups A and B, respectively (P < 0.0001). Interfering intestinal activity with varying degrees of intensity was seen in 20 patients in group A (49%) and in 5 patients in group B (13%; P < 0.001). CONCLUSIONS: These results suggest that filling of the stomach with some fluid and solid food before rest acquisition on the same day stress--rest protocol, because of its volume effect, may provide a high target-to-nontarget ratio for myocardial imaging and thus may reduce the frequency of intestinal artifacts. PMID- 11416746 TI - In-111 leukocyte accumulation in aberrant bone marrow after spinal fusion mimicking paraspinal infection. PMID- 11416747 TI - Detection of cerebrospinal fluid leak by dual-isotope spect with In-111 DTPA and Tc-99m HMDP. PMID- 11416748 TI - Pyonephrosis in a duplex kidney: diagnosis facilitated by skeletal scintigraphy. PMID- 11416749 TI - Diuretic renogram in a patient with a urinary diversion: avoiding a false positive diagnosis of obstruction with an indwelling catheter. PMID- 11416750 TI - Rotation anomaly in duplex kidney. PMID- 11416751 TI - Paget's disease of the ulna: a rare location. PMID- 11416752 TI - "Superscan" in an autosomal-dominant benign form of osteopetrosis. PMID- 11416753 TI - F-18 FDG positron emission tomography to differentiate diabetic osteoarthropathy from septic arthritis. PMID- 11416754 TI - Diagnosis of intraperitoneal urine leak in a patient with obstructive hydronephrosis using dynamic renal scintigraphy and radionuclide antegrade pyelography. PMID- 11416755 TI - Ga-67-positive and F-18 FDG-negative imaging in well-differentiated hepatocellular carcinoma. PMID- 11416756 TI - Hepatobiliary scintigraphy in an exacerbation of Dubin-Johnson syndrome. PMID- 11416757 TI - Global absence of pulmonary perfusion in a hypoplastic lung with pulmonary artery agenesis associated with immunoglobulin a deficit. PMID- 11416758 TI - Localization of the breast sentinel node after axillary node dissection with diversion of lymphatic drainage to internal mammary lymph nodes and the importance of delayed imaging. PMID- 11416759 TI - Multicentric carcinoid tumor of the ileum localized by radioguided laparotomy. PMID- 11416760 TI - Buttock granulomas: a consequence of intramuscular injection of Sandostatin detected by In-111 octreoscan. PMID- 11416761 TI - Tc-99m HMPAO and Tl-201 uptake patterns in a malignant undifferentiated orbital neoplasm. PMID- 11416762 TI - Ga-67 uptake in ischemic enteritis in a patient with Churg-Strauss syndrome. PMID- 11416763 TI - Pregnancy-induced uterine blush detected incidentally by three-phase bone scintigraphy. PMID- 11416764 TI - Scintigraphic appearance in Lysol-induced lung toxicity. PMID- 11416765 TI - Secretion of radioactivity in breast milk after Tc-99m DTPA aerosol and Tc-99m MAA lung imaging. PMID- 11416766 TI - False-positive Tl-201 SPECT in patients with RBBB. PMID- 11416768 TI - Maternal Prepregnancy Weight and Congenital Heart Defects in the Offspring. AB - To determine the relation between having an infant with a major heart defect and a mother's prepregnancy weight, we compared 1,049 Atlanta-area women who gave birth to liveborn or stillborn infants, each with a major heart defect, with 3,029 Atlanta-area women who gave birth to infants without birth defects. The infants of control women were randomly selected from birth certificates and were frequency-matched to the case group by race, birth hospital, and birth period from 1968 through 1980. After excluding diabetic mothers and adjusting for potential confounders, compared with average-weight women (body mass index 19.9 22.7), we found that underweight women (body mass index <16.5) were less likely to have a child with a major isolated heart defect [odds ratio (OR) = 0.64; 95% confidence interval (CI) = 0.43-0.97], whereas the OR was elevated among overweight or obese women (body mass index >26) (OR = 1.36; 95% CI = 0.95-1.93). Using average-weight women who did not take periconceptional multivitamins as the reference group, periconceptional multivitamin use was associated with a reduced OR for isolated heart defects among average-weight women (OR = 0.61, 95% CI = 0.36-0.99) and underweight women but not among overweight or obese women (OR = 1.69, 95% CI = 0.69-3.84). PMID- 11416769 TI - Our policy on policy. PMID- 11416770 TI - Policy emanating from epidemiologic data: what is the proper forum? PMID- 11416771 TI - Policy and science: should epidemiologists comment on the policy implications of their research? PMID- 11416772 TI - Epidemiology at the edge. PMID- 11416773 TI - Early life circumstances and late life Alzheimer's disease. PMID- 11416774 TI - Alcohol and coronary heart disease: can we learn more? PMID- 11416775 TI - Using census data and birth certificates to reconstruct the early-life socioeconomic environment and the relation to the development of Alzheimer's disease. AB - The early-life environment and its effect on growth and maturation of children and adolescents are associated with several adult chronic diseases, including Alzheimer's disease. Because it is not feasible to collect information prospectively over the average life span, methods to reconstruct the early-life environment of the aged are necessary to evaluate these associations. In a community-based case-control study conducted in the United States, we collected U.S. census records and birth certificates to reconstruct the early-life socioeconomic environment of each elderly subject. Information was found on 82% of the available Alzheimer's disease cases (239 of 292) and 87% of the available controls (245 of 282). We investigated risk of Alzheimer's disease associated with father's occupation, parental age, household size, sibship size, and birth order. Subjects whose fathers were unskilled manual workers or laborers were at higher risk for Alzheimer's disease (odds ratio = 1.80, 95% confidence interval = 1.19--2.73). The risk of Alzheimer's disease was increased with increasing number of people in the household. We also evaluated whether subjects with the apolipoprotein epsilon 4 allele (APOE epsilon 4), a strong genetic risk factor that is not a necessary cause or a sufficient cause by itself for the development of Alzheimer's disease, were at higher risk than subjects who did not carry this allele. Among subjects with the APOE epsilon 4 allele whose fathers held lower socioeconomic level occupations, the odds of developing Alzheimer's disease were higher (odds ratio = 2.35, 95% confidence interval = 1.07--5.16) compared with subjects without the allele (odds ratio = 1.40, 95% confidence interval = 0.78- 2.52). Subjects carrying the APOE epsilon 4 allele alone have a threefold increased risk of Alzheimer's disease (odds ratio = 3.17, 95% confidence interval = 1.99--5.04). Compared with subjects with neither risk factor, subjects with both the genetic and the environmental risk factors (household size of seven or more and father's occupation being manual) had a relatively high risk of Alzheimer's disease (odds ratio = 14.8, 95% confidence interval = 4.9--46). The data suggest that APOE epsilon 4 may modify the associations between father's occupation, other early-life environmental factors, and development of Alzheimer's disease in late life. PMID- 11416776 TI - Coronary heart disease risk reduction in a predominantly beer-drinking population. AB - Moderate alcohol consumption protects against coronary heart disease, but there is ongoing debate about whether the effects and the underlying mechanisms differ by type of alcoholic beverage. In our case-control study from Germany, we assessed and compared the effect of alcohol consumption from various sources on risk of coronary heart disease and its underlying mechanisms. The sample included 312 patients with clinically stable, angiographically confirmed coronary heart disease and 479 healthy controls. After control for potential confounding factors, the odds ratio for coronary heart disease was 0.55 (95% confidence interval = 0.37--0.83) for drinkers compared with nondrinkers. Risk reduction was particularly strong among the majority of study participants who reported drinking predominantly or exclusively beer. Alcohol consumption from various sources was consistently associated with a more favorable profile of lipids and hemostatic factors. Nevertheless, a strong inverse association persisted between beer drinking and coronary heart disease even after control for these "intermediate variables." Our results support suggestions that the protective effect of moderate alcohol consumption against coronary heart disease is mediated in part by beneficial effects of ethanol on lipids and hemostatic factors. Additional mechanisms may account for the strong protective effect of moderate beer consumption. PMID- 11416777 TI - Residential radon and lung cancer among never-smokers in Sweden. AB - In this study, we attempted to reduce existing uncertainty about the relative risk of lung cancer from residential radon exposure among never-smokers. Comprehensive measurements of domestic radon were performed for 258 never-smoking lung cancer cases and 487 never-smoking controls from five Swedish case-control studies. With additional never-smokers from a previous case-control study of lung cancer and residential radon exposure in Sweden, a total of 436 never-smoking lung cancer cases diagnosed in Sweden between 1980 and 1995 and 1,649 never smoking controls were included. The relative risks (with 95% confidence intervals in parentheses) of lung cancer in relation to categories of time-weighted average domestic radon concentration during three decades, delimited by cutpoints at 50, 80, and 140 Bq m(-3), were 1.08 (0.8--1.5), 1.18 (0.9--1.6), and 1.44 (1.0--2.1), respectively, with average radon concentrations below 50 Bq m(-3) used as reference category and with adjustment for other risk factors. The data suggested that among never-smokers residential radon exposure may be more harmful for those exposed to environmental tobacco smoke. Overall, an excess relative risk of 10% per 100 Bq m(-3) average radon concentration was estimated, which is similar to the summary effect estimate for all subjects in the main residential radon studies to date. PMID- 11416778 TI - Kidney disease and arthritis in a cohort study of workers exposed to silica. AB - Silica exposure has been associated with kidney disease and rheumatoid arthritis; an autoimmune mechanism has been proposed. Approximately 2 million people are occupationally exposed to silica in the United States, 100,000 at more than twice the National Institute for Occupational Safety and Health recommended exposure limit of 0.05 mg/m(3). We examined renal disease morbidity and mortality, as well as arthritis mortality, in a cohort of 4,626 silica-exposed workers in the industrial sand industry (an industry previously unstudied). We compared the cohort with the U.S. population and also conducted internal exposure-response analyses using a job-exposure matrix based on more than 4,000 industrial hygiene samples. We found excess mortality from acute renal disease [standardized mortality ratio (SMR) = 2.61, 95% confidence intervals (95% CIs) = 1.49--4.24; 16 deaths], chronic renal disease (SMR = 1.61, 95% CI = 1.13--2.22; 36 deaths), and arthritis (SMR = 4.36, 95% CI = 2.76--6.54; 23 deaths) on the basis of multiple cause mortality data, which considered any mention of disease on a death certificate. Linking the cohort with the U.S. registry of end-stage renal disease for the years 1977-1996, we found an excess of end-stage renal disease incidence (standardized incidence ratio = 1.97, 95% CI = 1.25--2.96; 23 cases), which was highest for glomerulonephritis (standardized incidence ratio = 3.85, 95% CI = 1.55--7.93; 7 cases). We found increasing end-stage renal disease incidence with increasing cumulative exposure; standardized rate ratios by quartile of cumulative exposure were 1.00, 3.09, 5.22, and 7.79. A positive exposure-response trend was also observed for rheumatoid arthritis on the basis of death certificate data. These data represent the largest number of kidney disease cases analyzed to date in a cohort with well-defined silica exposure and suggest a causal link between silica and kidney disease. Excess risk of end-stage renal disease due to a lifetime of occupational exposure at currently recommended limits is estimated to be 14%, above a background end-stage renal disease risk of 2%. PMID- 11416779 TI - Effects of ambient air pollution on daily mortality in a cohort of patients with congestive heart failure. AB - This study was planned to test the hypothesis that patients with congestive heart failure are more susceptible to the harmful effects of ambient air pollution than the general population. We used both general additive Poisson regression and the case-crossover approach to test the hypothesis. The effect of air pollution on daily mortality of patients with congestive heart failure among residents of Seoul, South Korea, during the period 1994--1998 was compared with that of the general population in the same area and the same period. The odds ratios and 95% confidence intervals (95% CIs) estimated from general additive models in the general population for an interquartile range increase of particulate matter less than 10 microm in diameter (42.1 microg/m(3)), carbon monoxide (0.59 ppm), nitrogen dioxide (14.6 ppb), sulfur dioxide (9.9 ppb), and ozone (20.5 ppb) were 1.014 (95% CI = 1.006- 1.022), 1.022 (95% CI = 1.017- 1.029), 1.021 (95% CI = 1.014- 1.029), 1.020 (95% CI = 1.012--1.028), and 1.010 (95% CI = 1.002--1.017), respectively. The estimated effects appeared larger among the congestive heart failure patients than among the general population (2.5 approximately 4.1 times higher depending on the pollutants). The results from the case-crossover analysis were similar. The finding of a stronger association in the patients with congestive heart failure reinforces the evidence that a harmful effect of air pollution is mediated by cardiovascular mechanisms. PMID- 11416780 TI - Dietary folate intake, alcohol, and risk of breast cancer in a prospective study of postmenopausal women. AB - Low B-vitamin intake may increase risk of breast cancer through decreased DNA repair capacity. Alcohol intake increases risk for breast cancer, with evidence from prospective studies of an interaction between alcohol and folate. We explored dietary intake of folate and other B vitamins with risk of breast cancer in a cohort study of 34,387 postmenopausal women. To measure diet, we mailed a food frequency questionnaire; we estimated nutrient intakes and categorized them into four levels: <10th, 11th-30th, 31st-50th, and >50th percentiles. Through 12 years of follow-up, we identified 1,586 cases of breast cancer in the cohort at risk. We estimated relative risks (RRs) and 95% confidence intervals (CIs) through Cox regression models adjusted for age, energy, and other risk factors. Women in the lowest 10th percentile of folate intake from diet alone were at modestly increased risk of breast cancer relative to those above the 50th percentile: RR = 1.21 (95% CI = 0.91--1.61). We examined the joint association of folate intake and alcohol use on risk of breast cancer, with the reference group defined as women with high folate (>50th percentile) and no alcohol use. The RRs of breast cancer associated with low dietary folate intake were 1.08 (95% CI = 0.78--1.49) among nondrinkers, 1.33 (95% CI = 0.86--2.05) among drinkers of < or = 4 gm per day, and 1.59 (95% CI = 1.05--2.41) among drinkers of > 4 gm per day. These results suggest that the risks of postmenopausal breast cancer may be increased among women with low intakes of folate if they consume alcohol containing beverages. PMID- 11416781 TI - Postmenopausal hormone use, screening, and breast cancer: characterization and control of a bias. AB - Previous investigators have suggested that screening-related biases may explain associations between postmenopausal hormone use and breast cancer. To investigate these biases, we studied postmenopausal women in the Nurses' Health Study from 1988 to 1994. Hormone use is associated with increased subsequent screening. Among women not screened in the previous 2 years, the probability difference, comparing current hormone users with others, for having mammography in the following 2 years is 19.5%; among women previously screened, the difference is 4.9%. These differences persist after control for other factors. If the increase in screening is causal, screening by mammogram could be intermediate in the causal pathway to breast cancer diagnosis. To deal with this problem, we restrict attention to a subset of the cohort in which the effect of postmenopausal hormone use on screening is small (women previously screened). In this subset, the rate ratio comparing breast cancer rates among current postmenopausal hormone users with others is 1.28. In a sensitivity analysis, the bias could not by itself plausibly account for the associations in our data. Our data provide evidence of an association between postmenopausal hormone use and breast cancer that is not solely the product of a detection bias. PMID- 11416782 TI - Increased risks of circulatory diseases in late pregnancy and puerperium. AB - We studied a nationwide Swedish cohort with 654,957 women who had 1,003,489 deliveries from 1987 through September 1995 to assess late pregnancy and puerperal risks of circulatory diseases. We used standardized incidence rate ratios to calculate relative risks [with 95% confidence intervals (CIs)]. Compared with unexposed (nonpregnant and early pregnant) women, relative risks of venous thrombosis and pulmonary embolism during the third trimester were 6.7 (95% CI = 5.7--7.8) and 2.7 (95% CI = 1.7--4.2), respectively. Around delivery (from 2 days before to 1 day after delivery), the relative risks of all assessed circulatory diseases were dramatically increased: venous thrombosis, 115.1 (95% CI = 96.4--137.0); pulmonary embolism, 80.7 (95% CI = 53.9--117.9); subarachnoid hemorrhage, 46.9 (95% CI = 19.3--98.4); intracerebral hemorrhage, 95.0 (95% CI = 42.1--194.8); cerebral infarction, 33.8 (95% CI = 10.5--84.0); and myocardial infarction, 27.0 (95% CI = 0.6--180.0). During the rest of the first 6 weeks postpartum, the risks declined but were still substantially increased for all diseases, with the exception of subarachnoid hemorrhage. The results suggest that the increased risk for circulatory diseases related to pregnancy is mainly confined to a few days around delivery. PMID- 11416783 TI - Recall bias in a case-control surveillance system on the use of medicine during pregnancy. AB - It is important to study possible teratogenic effects of drugs used during pregnancy. Many studies of this type rely upon case-control designs in which drug intake is recalled by the mothers after having given birth. Recall bias in this situation may lead to spurious associations. We looked for indicators of recall bias by comparing self-reported drug intake with medically notified intake for specific diseases in the Hungarian Case-Control Surveillance System of Congenital Abnormalities, which includes 22,865 cases with congenital abnormalities and 39,151 controls. Recall error was present, especially for drugs used for a short time period. Furthermore, the timing of drug intake was reported slightly closer to the time of interview for cases compared than for controls. Severe or visible congenital abnormalities did not appear to be more conducive to recall bias than other abnormalities under study. A case-control surveillance system of this type may frequently cause spurious associations, with biased odds ratios up to a factor of 1.9. PMID- 11416784 TI - Society, physical health and modern epidemiology. PMID- 11416785 TI - Antinomy and the S(B) model for SIDS. PMID- 11416786 TI - Colorectal cancer among users of aspirin and non-steroidal antiinflammatory drugs. PMID- 11416787 TI - Pooled analysis of magnetic fields, wire codes, and childhood leukemia. PMID- 11416788 TI - Reporting of attributable risk in epidemiologic studies. PMID- 11416789 TI - Minimum fully effective dose of folic acid for the prevention of neural tube defects. PMID- 11416790 TI - [MRI of aortic coarctation: impact in daily practice]. PMID- 11416791 TI - [Multislice helical CT: principles, applications]. AB - The real innovation of multislice helical CT is its technology of detectors. These detectors may be isotropic, anisotropic or hybrid according to the manufacturers. Whatever the concept in terms of geometric resolution or evolutivity, all of them provide similar implementations and image quality. Data transfer and computer performances are as much as eight times higher than those of single slice CTs. Large pitches are routinely available. The speed of workstation softwares has been dramatically increased. Acquisition times have also been reduced like on new single-detector CTs. According to the machine, a 360 degrees rotation of the tube is obtained in 0.5 to 0.8 sec. Evolution more than revolution, there is need for further clinical expertise to know whether this new technology is able to substitute for other imaging modalities. In the meantime, what is sure is that it will generate new clinical applications. PMID- 11416792 TI - [The masseter: an enigmatic muscle]. AB - PURPOSE: To try and explain the frequency of pathologic processes involving the masseter muscle. MATERIALS AND METHODS: Anatomical and histological evaluation of the masseter muscle and its partial continuity with the medial pterygoid muscle. Retrospective evaluation of 21 cases of pathology of the masseter muscle imaged by CT and/or MRI. RESULTS: Two main groups of pathology are identified: benign masseter hypertrophy and hemodynamically inactive vascular malformations. Other etiologies are quite varied. Some are secondary to processes involving the nearby mandible and parotid gland. Others, including tumors, involved the masseter muscle in a non specific fashion. CT and especially MRI were quite valuable with accurate diagnosis in 17/21 cases, two of which with the help of clinical data. CONCLUSION: The main question regarding the preferential involvement of the masseter muscle over nearby muscles remains unanswered. PMID- 11416793 TI - [Thoracic aortic coarctation: MRI evaluation and follow-up]. AB - PURPOSE: Report our experience in the evaluation and follow-up of thoracic aortic coarctation with MRI and describe its role to estimate trans-stenotic flow. MATERIAL AND METHODS: 43 MR examinations were performed in 30 patients (age range 15 days to 73 years) referred to our institution in the last 7 years. RESULTS: MRI visualized the ascending, horizontal and descending portions of the aorta and the supra-aortic vessels in 42/43 patients. MRI clearly identified preoperatively an aortic coarctation in 11/12 cases confirmed by surgery. Postoperatively MRI depicted 4 restenosis and one aneurysm. In 5 patients MRI demonstrated pseudo coarctation. A significant correlation was established between the maximal trans stenotic pressure gradient when measured by Doppler US or angiography and the size of the signal void measured on cine-MR images (r=0.72; p<0.01). CONCLUSION: MRI is a reliable non invasive investigation method for the diagnosis and semi quantitative evaluation of aortic coarctation particularly when colour Doppler US is not satisfactory. PMID- 11416794 TI - [Value of high frequency (20mhZ) and doppler ultrasound in the diagnosis of pigmented cutaneous tumors]. AB - PURPOSE: Diagnosis of pigmented skin tumors is often difficult. The aim of the present study is to evaluate high frequency ultrasound and Doppler ultrasound in the localization and the characterization of these tumors. MATERIAL AND METHODS: One hundred and seventy six pigmented skin tumors including melanocytic and non melanocytic, benign and malignant lesions have been examined before resection using high frequency ultrasound (20MHz probe) and Doppler ultrasound (13MHz probe Doppler frequency: 7MHz). Imaging data have been correlated with histological data, available in all cases. RESULTS: Tumors seen at ultrasound were hypoechoic lesions, some of which, particularly malignant melanocytic tumors were vascularized. Precise ultrasonographic location of the tumor was correlated to its histological location: epidermis for purely epidermal proliferations (seborrheic keratoses), superficial dermis for dermal proliferations originating from epidermis (benign dermal nevi, melanomas during vertical growth phase, basal cell carcinomas), middle and deep dermis for dermal proliferations originating from dermis (fibrous histiocytomas, angiomas). CONCLUSION: Ultrasound may help in assessing positive and differential diagnosis of pigmented skin tumors as it precises, before histological examination, the exact location of the lesion within the different skin layers and its origin. As far as malignant melanocytic tumors are concerned, high frequency ultrasound and Doppler ultrasound appear promising in diagnosis algorithm and pronosic of these lesions. PMID- 11416795 TI - [Preoperative management of hip osteochondritis: comparison of MRI and arthrography with abduction views]. AB - PURPOSE: Comparison between conventional arthrography and MRI in the preoperative work up of Legg-Calve-Perthes disease. MATERIAL: ans methods. We studied 17 children (18 hips) who had arthrography and MRI (open-magnet configuration) the same day. Coronal and sagittal T1 weighted were realised with the hip in neutral position. Additional coronal T1 weighted with abduction of the hip were obtained. Two readers analysed individually MRI and arthrographic findings: shape of femoral head, index of Heyman (antero-external covering), came effect, recentering of femoral head and quality of examimation. RESULTS: No statistical difference between MRI and arthrography could be noted-exception made for the index of Heyman. CONCLUSION: MRI can replace arthrography in Legg-Calve-Perthes disease. PMID- 11416796 TI - [Os subfibulare in children secondary to an osteochondral fracture]. AB - PURPOSE: The understanding of the os subfibulare in childhood. MATERIAL AND METHODS: A retrospective study of radiographs following for indirect ankle injury in childhood over a period of two years. RESULTS: The daily use of an oblique view during the exploration of ankle sprains in emergency has allowed us to image 12 small and linear chiplike fractures of the fibula by avulsion of the antero medial part of the malleolar's tip where the talo-fibular collateral ligament is attached. Two children were reviewed one year after for a recurrent sprain: a larger and rounder fragment of bone has taken the place of the first piece of bone. CONCLUSION: Symptomatic ossicles of the lateral malleolus in children is frequently if not always the consequence of an avulsion of the tip of the fibula. The outcome of these children may be a localised tenderness and recurrent episodes of sprains for minimal injury due to the weakness of the lateral ligament. PMID- 11416797 TI - [Retroperitoneal complications of Erdheim-Chester disease]. AB - Erdheim-Chester disease is a non-langerhans cell histiocytosis. It is a rare and plurifocal disease. Retroperitoneal involvement manifests as a mass associated with fibrosis, which is well visualized on CT scan and MRI. This disease is characterized by its potential to involve the whole retroperitoneum. We report a case of this disease that developed over twenty years, consisting of renal arteries stenosis, bilateral ureteral stenosis and evolutive adhesive capsulitis. PMID- 11416798 TI - [Pulmonary embolization from migration of sclerotherapy material]. AB - We present a case of pulmonary embolism which resulted from the migration of an oesophageal variceal sclerosing agent. Despite its frequency, this complication is rarely described in the literature. PMID- 11416799 TI - [Bilateral ureteral pseudodiverticulosis with transitional cell tumour]. AB - We present with a case of bilateral ureteral pseudodiverticulosis associated with a transitional cell tumour in the bladder. From a pathogenic point of view, there is currently a trend to consider reactive hyperplasia of transitional cell epithelium as secondary to chronic inflammatory processes which would form such pseudodiverticula. On account of that association it is advisable to perform follow-up tests on these patients to enable us to reach an early diagnosis of these tumours. PMID- 11416800 TI - [Unusual radiologic signs of bone echinococcosis in children]. AB - The authors report 2 cases of 2 children aging 11 and 12-years-old presenting hydatitosis in the upper extremity of 2 long bones femur and tibia. In 2 cases, radiological appearances were atypical. In the femur, plain radiographs revealed multiple lucencies, irregular, located in the superior extremity with a periostal reaction. The CT scan confirmed these data and showed a cortical rupture, extension to soft tissue with contrast enhancement. In the tibia, plain radiographys showed delimitated lucencies, confluent with a rim condensation in the proximal epiphyseal-metaphyseal area. Echography and CT are helpful when radiological appearances are unusual. PMID- 11416801 TI - [Does the upper respiratory tract high-resistance syndrome exist?]. PMID- 11416802 TI - [Chemoprevention perspectives for bronchial cancer]. AB - Lung cancer, with a high incidence and a 14% survival rate at 5 years is the leading cause of death in France. Because of past and present smoking habits in the French population and the lack of real expectations for significant therapeutic progress within a short or mid-term delay, the only reasonable way to try to limit the predictable hecatomb in the next 2 decades is to reduce exposure to the main risk factors (tobacco smoke, asbestos.), implement an early and effective (radiographic and/or endoscopic) screening system, and/or determine an active chemoprevention scheme. The principle of chemoprevention is based on the fundamental concept that since lung cancer develops through several stages, subjects exposed to risk factors could be given a compound or compounds counteracting the deleterious effect of carcinogenic substances on DNA and/or blocking the subsequent cascade of molecular events. Two families of products have been considered as potential chemoprevention agents: antioxidants (selenium, beta-caroten, vitamin E, and N-acetyl-cystein) and vitamin A and its analogs. Unfortunately, despite promising experimental data, large-scale clinical trials have not evidenced any protective effect of these compounds that have even been observed to produce opposing effects. To date, no chemopreventive substance can be reasonable proposed for subjects at risk. Due to the lack of proof of the efficacy of the different screening systems proposed, the only preventive action with proven efficacy is to limit exposure to risk factors. All health care givers must actively participate in the fight against active and passive smoking. PMID- 11416803 TI - [Lung autofluorescence. Preliminary study of two systems without laser illumination or photosensitization]. AB - When illuminated with a certain wavelength, lung tissue emits a weak fluorescence. The fluorescence from normal tissue is different from that from diseased tissue. This technique can thus be used for early diagnosis of precancerous lesions. Technical manipulations are required to amplify the fluorescence signal. Numerous studies have used the LIFE (Lung Imaging Fluorescent Endoscope) to study lung diseases. This system of detection uses a laser helium-cadmium excitation source; the principal photosensitizing agents are hematoporphyrin derivatives. The cost, bulkiness and secondary effects have limited diffusion of this type of photodynamic diagnostic system. We have used 2 systems (SAFE 1000 Pentax and D-LIGHT Storz) successively in a prospective study to analyze autofluorescence of lung tissue without laser illumination or photosensitization. Fifty-four patients were selected for their risk factors. We were able to detect one case of severe dysplasia but did not identify any case of carcinoma in situ. Among 18 cases, 16 were evident with white light. Two were detected with autofluorescence used to orient biopsies while white light had only revealed a common inflammatory aspect. Thirty-nine low-grade lesions (metaplasia, dysplasia) were found. The systems are easy to use. Lower cost would allow more widespread use of these systems currently reserved for risk populations. These techniques broaden the scope of interventional endoscopy. PMID- 11416804 TI - [Respiratory rehabilitation in patients with bronchial asthma and chronic obstructive pulmonary disease (COPD) in Kinshasa]. AB - A number of studies in western countries have shown that respiratory and physical rehabilitation of patients with chronic obstructive pulmonary disease (COPD) only has a minimal effect on respiratory function but can significantly improve physical capacity. The aim of our study was to apply these methods to patients in Kinshasa, Democratic Republic of Congo. We treated 38 patients (20 women) who had bronchial asthma (n=14) or COPD (n=24). These ambulatory patients were treated in two different hospitals in Kinshasa, the university hospital and the general hospital, three times per week for twelve weeks. The patients were treated with kinestherapy and inhaled bronchodilator drugs (salbutamol, and/or ipratropium bromide with a boreal nebulizer) as well as bronchial hygiene and performed breathing exercises on a Bodyguard Ergometer 990 with walking, running and climbing steps until exhaustion. After the rehabilitation program FEV1 increased from 1.37 +/- 0.62 (50% expected) to 1.54 +/- 0.69 (56% expected) (p<0.01). The same trend was observed for walking distance (from 644 +/- 459 m to 1213 +/- 569 m, p<0.001) and for maximal power developed on the cycloergometer (from 45 +/- 20 w to 73 +/- 37 w, p<0.001). In contrast, the maximal work load performed during climbing steps (from 106 +/- 44 w to 115 +/- 23 w) did not improve significantly. COPD patients improved their FEV1 significantly compared with asthma patients. Our study show that pulmonary rehabilitaion increase the level of spontaneous physical activity. The pulmonary rehabilitation program changes the quality of life of COPD patients who are able to move about better for longer periods of time, have a longer walking distance, and improved physical activity level. PMID- 11416805 TI - [Aspergillous bronchitis in an immunocompetent patient]. AB - We report a case of aspergillous bronchitis in an immunocompetent patient, recalling the clinical signs, laboratory findings and therapeutic management of this uncommon bronchopulmonary disorder related to aspergillus. PMID- 11416806 TI - [Pulmonary metastasis from a glioblastoma. A case report]. AB - Glioblastoma is a primary malignant astrocyte tumor of the central nervous system. Extraneural metastasis is uncommon. We report a case of spontaneous lung metastasis from a glioblastoma without prior surgery. Positive diagnosis was achieved from histology and immunohistochemistry PMID- 11416807 TI - [Mediastinal adenopathies and presence of anti-cytoplasmic antibodies of diffuse type polynuclear neutrophils]. AB - We report a case of sarcoidosis with mediastinal and pulmonary localizations associated with diffuse antineutrophil cytoplasmic antibodies (c-ANCA). This led to discuss the differential diagnosis with vascularitis and the specificity of c ANCA in Wegener's granulocytosis. PMID- 11416808 TI - [Analgesic effect of Pamidronate on bone pain in patient with hypertrophic pulmonary osteoarthropathy]. PMID- 11416809 TI - Hanging tough. PMID- 11416810 TI - Ethics development for clinical faculty. PMID- 11416811 TI - Deep change. PMID- 11416812 TI - A commentary on comments. PMID- 11416813 TI - The fading of nursing leadership. AB - Based on recent BSN student clinical observations and evaluations from inpatient hospital units to which they were assigned, we were concerned about the absence of nursing leadership in these settings and the impact this absence has on our profession. In this opinion article, we explore how nursing leadership is being influenced by the structure of health care institutions and by an anti intellectual stance among members of the profession. We discuss how these factors are influencing the current state of our profession and recommend a course of action to ensure the professional presence of nursing in the future. We also encourage readers to think about what they can do to prevent the fading of nursing leadership. PMID- 11416814 TI - A new curriculum for a new millennium. AB - In the last few years of the 20th century, many factors converged at Auburn University School of Nursing, giving faculty the opportunity to explore beliefs about curriculum and the nursing profession's role in the 21st century. The result of this opportunity was the creation of a new curriculum that brings to life the conviction that nursing is not bound by the 4 walls of a hospital, but is a humanistic discipline that can be practiced anywhere. This article explains the intersection of variables that influenced the development of a new curriculum, including the call by nursing leaders for educators to drastically change their approach to the education of students. We describe the process of developing a community-based curriculum, with innovative examples of clinical and theory experiences on a semester-by-semester basis. Strategies to address obstacles to implementation of a community-based curriculum are included. PMID- 11416815 TI - Enhancing traditional, televised, and videotaped courses with Web-based technologies: a comparison of student satisfaction. AB - BACKGROUND: Varied distance learning strategies can be used to deliver nursing courses, including interactive television, videotape, and Web-based approaches. PURPOSES: (1) To assess student assess student satisfaction with a critical care elective course offered simultaneously via traditional and distance learning formats in which Web-based strategies were added, and (2) to compare satisfaction of students taking the traditional course versus those taking the class via distance technology. METHODS: Students (n = 113) who took the course during the spring 1998 and 1999 semesters completed a teacher-constructed evaluation at the end of the semester. FINDINGS: Mean ratings on the evaluation were positive. Ratings of interaction, communication with instructor, and facilitation of learning were higher from students who took the traditional course. CONCLUSIONS: The application of Web-based technologies may be one factor for the overall course satisfaction. However, it is important to continue to evaluate strategies that work best for students taking courses via distance technology. PMID- 11416816 TI - End-of-life competencies and the NCLEX-RN examination. AB - With the aging of the population, health care workers must be prepared to deal with clients and families facing end-of-life (EOL) issues. Nurses, as one of the primary providers of care and the largest professional group to provide care, can make a significant impact on EOL care. This article uses 15 nursing competencies that have been identified as important in providing high-quality care to clients and the families of clients at the end of life and delineates where these competencies are addressed in the 1998 NCLEX-RN Test Plan. Available resources that provide information on EOL content are mentioned as well as how to become involved in the NCLEX item development process. PMID- 11416817 TI - Baccalaureate nursing education development in 2 Baltic countries: Outcomes 10 years after initiation. AB - BACKGROUND: Baccalaureate nursing education began in the Baltic countries of Latvia and Lithuania ten years ago to prepare nursing teachers and leaders. PURPOSE: The purpose of the study was to assess the outcomes of the initial baccalaureate programs for nurses in Latvia and Lithuania from the perspective of program graduates. METHOD: The method was an alumni survey developed by the authors in English and translated into the respective languages. DISCUSSION: Most of the Latvian and Lithuanian alumni respondents were staff nurses on entering the baccalaureate programs, but after graduation were making a contribution as nursing leaders and teachers. CONCLUSIONS: The initial baccalaureate programs for nurses have been followed by increasing opportunities for nurses to pursue graduate study and careers in and outside of nursing. The infusion of prepared local nursing faculty to replace physician faculty and foreign short-term guest faculty promises continued development of nursing education. PMID- 11416818 TI - The nature of advanced practice nursing. AB - In attempting to define "advanced practice," we argue that nursing as such is teleological or goal-directed with those goals being defined by the patient or client in interaction with the nurse. In helping the patient meet identified goals, the nurse requires 2 kinds of knowledge-general and particular. General includes theory (know what/why), pattern recognition (know what), and practical knowledge (know how). Particular (know who) is personal knowledge about the patient. The advanced practice nurse, by virtue of graduate education, is able to move beyond the familiar and experientially learned. He or she makes a deliberate attempt to situate self in a dialectic between general and particular knowledge in such a way that the interplay opens possibilities. Knowing when a particular action would be most helpful is defined as practical wisdom. We argue that a highly developed sense of practical wisdom is the hallmark of advanced practice. PMID- 11416819 TI - President's message: an era for collaboration. PMID- 11416820 TI - Chiropractors and return-to-work: the experiences of three Canadian focus groups. AB - OBJECTIVES: To explore the views of chiropractors about timely return-to-work in treating patients with musculoskeletal injuries, to identify the approaches used by chiropractors when treating injured workers with musculoskeletal disorders, and to learn about chiropractors' perspectives on the barriers and facilitators of successful return-to-work. DESIGN: Qualitative study of 3 focus groups of chiropractors. METHODS: Focus groups of 8 to 11 chiropractors were conducted in 3 large Canadian cities. The selected participants were experienced in treating patients with occupational musculoskeletal injuries. Standard questions were used to collect data. The data from each focus group were coded and analyzed separately and then considered in relation to each other. RESULTS: The participants indicated that timely return-to-work depends on patients' characteristics, severity of injury, clinical progress, the availability of work accommodation, and clinical judgment. The chiropractors commented that their treatment of injured workers rests on their strength in diagnosis and treatment and on providing patient-centered care. Positive human relations within workplaces and the ability to accommodate the work of an injured worker were described as important in return-to-work programs. The participants believed that a bias against chiropractic is present within the medical profession and workers' compensation boards. They viewed this bias as an important barrier when assisting their patients to successfully return to work. CONCLUSION: The broad approaches described by the participating chiropractors to return injured workers to work are consistent with those proposed in evidence-based practice guidelines. Better communication among chiropractors, medical doctors, and workers' compensation boards would likely decrease interprofessional tensions and improve the recovery of workers with musculoskeletal injuries. PMID- 11416821 TI - Posterior disk displacement: morphologic assessment and measurement reliability lumbar spine. AB - BACKGROUND: Magnetic resonance imaging is often used to assess for disk displacement after manipulation, but limited information about the true incidence of iatrogenic herniations exists. To design a study that evaluates for a causal relationship, preliminary data must be obtained relating to the size of different types of disk displacement. The reliability of chiropractic radiologists in assessing disks and a comparison of different measuring devices should also be evaluated. OBJECTIVE: To identify average measurements for normal and displaced disks and to assess the reliability of measurements by chiropractic radiologists. STUDY DESIGN: Intraobserver and interobserver reliability study assessing disk displacement on magnetic resonance scans. METHODS: Three evaluators assessed the disks on 122 magnetic resonance scans from two imaging centers. Six categories were graded, and digitizer and ruler measurements were compared. Forty-four scans were reassessed for intraobserver agreement. Intraobserver and interobserver variations were measured with intraclass correlation coefficient and kappa statistical analysis. Measurement device correlation was assessed with Pearson's r. RESULTS: Clear size differences between different types of disk displacement were noted. Interexaminer measurement reliability was 0.78 to 0.84. Agreement concerning the presence of disk displacement was 85% (kappa = 0.68), and the classification of disk displacements was 76% (kappa = 0.60). Intraexaminer measurement reliability was 0.40 to 0.49. Intraexaminer agreement concerning the presence of disk displacement was 76% (kappa = 0.52), and the classification of disk displacements was 62% to 69% (kappa = 0.38 to 0.46). Normal versus bulged disk distinctions demonstrated the most disagreement. The ruler and digitizer correlation coefficient was 0.968. CONCLUSIONS: Different disk types demonstrated distinct size averages. Interexaminer agreement was good concerning disk assessment and measurements. Intraexaminer agreement was lower than expected. A millimetric ruler is an acceptable alternative to digital measurement devices. PMID- 11416822 TI - Errors in precise examiner head placement during cervical range-of-motion measurements. AB - OBJECTIVE: To quantify errors associated with examiner-assisted neutral head placement in cervical range of motion measurements in normal subjects and to investigate the influence of these errors on range of motion measurements. DESIGN: Repeated-measures design with cervical range of motion and errors in placement measured in 20 volunteers with no symptoms with the OSI CA-6000. METHODS: Examiner placement of head position was achieved with inclinometers and triangulation. Subjects estimated pain experienced during measurements with numeric pain scales. Angular data around 3 axes were analyzed with descriptive statistics. Possible correlations between errors and other variables were investigated. RESULTS: Drift, defined as displacement from original head positioning at first data acquisition and before initiation of motion, was negligible (+/-0.8 degrees ). Standard errors in neutral head placement ranged from 1.0 degrees in axial rotation to 3.2 degrees in flexion/extension. Within trial variability of neutral position did not correlate with between-trial differences in ranges of motion. CONCLUSION: Head position errors were not the primary sources of variability for between-trial measurements of cervical range of motion. The largest errors were in flexion/extension, and least, in axial rotation. Neutral position errors up to approximately 5 degrees for lateral bending, 3 degrees for rotation, and 9 degrees for flexion and extension fall within 95% CI and are the recommended lower limits for significant changes in clinical settings. PMID- 11416823 TI - The health information brochure: a useful tool for chiropractic practice? AB - BACKGROUND: It has been suggested that clinicians should be looking at new ways to enhance their patients' self-care. Patient education is one strategy that primary providers may use. OBJECTIVE: This study investigates the format in which patients would like to pursue their health education within the chiropractic clinic. METHODS: An exploratory study of chiropractic patients was undertaken to investigate patients' preferred health education formats, their commitment to pursuing health objectives, and their literacy level. Purposive sampling of 9 Australian chiropractic clinics was undertaken. Convenience sampling of patients attending these clinics resulted in 102 patients participating. Participants completed a questionnaire. A research assistant was available to clarify any questions. Data were collected and collated. A Likert scale was used to capture responses to questions ascertaining patient opinions. RESULTS: Patients considered health the most important of the life objectives listed; however, they preferred spending time with family to undertaking health- and fitness-promoting activity. More chiropractic patients opted for health information brochures than health promotion classes, personally supervised self-care programs, or practitioner-supervised self-care contracts. Patient literacy levels varied within and between clinics. CONCLUSIONS: Brochures may provide a definitive health information tool for chiropractors who limit their clinical role to primary contact and a helpful adjunct to patient education for chiropractors committed to a primary care role. However, care should be taken to select brochures consistent with the patients' literacy level. Tips for selecting and preparing suitable brochures are provided. The discrepancy between how greatly patients value health and how they prefer spending their time may have implications for successful behavior change. Brochures may not alone constitute adequate practitioner involvement. PMID- 11416824 TI - Medically supervised water-only fasting in the treatment of hypertension. AB - BACKGROUND: Hypertension-related diseases are the leading cause of morbidity and mortality in industrially developed societies. Although antihypertensive drugs are extensively used, dietary and lifestyle modifications also are effective in the treatment of patients with hypertension. One such lifestyle intervention is the use of medically supervised, water-only fasting as a safe and effective means of normalizing blood pressure and initiating health-promoting behavioral changes. METHODS: One hundred seventy-four consecutive hypertensive patients with blood pressure in excess of 140 mm Hg systolic, 90 mm Hg diastolic (140/90 mm Hg), or both were treated in an inpatient setting under medical supervision. The treatment program consisted of a short prefasting period (approximately 2 to 3 days on average) during which food consumption was limited to fruits and vegetables, followed by medically supervised water-only fasting (approximately 10 to 11 days on average) and a refeeding period (approximately 6 to 7 days on average) introducing a low-fat, low-sodium, vegan diet. RESULTS: Almost 90% of the subjects achieved blood pressure less than 140/90 mm Hg by the end of the treatment program. The average reduction in blood pressure was 37/13 mm Hg, with the greatest decrease being observed for subjects with the most severe hypertension. Patients with stage 3 hypertension (those with systolic blood pressure greater than 180 mg Hg, diastolic blood pressure greater than 110 mg Hg, or both) had an average reduction of 60/17 mm Hg at the conclusion of treatment. All of the subjects who were taking antihypertensive medication at entry (6.3% of the total sample) successfully discontinued the use of medication. CONCLUSION: Medically supervised water-only fasting appears to be a safe and effective means of normalizing blood pressure and may assist in motivating health-promoting diet and lifestyle changes. PMID- 11416825 TI - Postural dynamics: clinical and empirical implications. AB - OBJECTIVE: To provide a rationale for the examination of posture from a dynamic (behavioral) perspective and to relate the vertebral subluxation to postural instability and motion sickness via inefficiency. DATA COLLECTION: A manual search of available reference texts and a computer search of literature from Index Medicus, PsycINFO, and ISI Science Citation Index Expanded were collected with an emphasis on postural dynamics, vertebral subluxation, and motion sickness. RESULTS: Evidence linking behavioral and health research has emerged from the study of posture and postural dynamics. Studies examining the relation between postural control and motion sickness have shown that motion sickness is preceded and predicted by postural instability. Motion sickness is characterized by maladaptive response to unusual motion events. The symptoms are nonspecific and variable. Although the Postural Instability theory of motion sickness predicted that instability should precede sickness, it did not make any claims regarding the symptoms associated with it. Chiropractic literature has emphasized the effects of vertebral subluxation on neurologic dysfunction. Vertebral subluxation is a condition that is postulated to interfere with neurologic processes and may influence organ system function and general health. As in the case of motion sickness, symptoms are nonspecific and variable (and in some instances the person may have no symptoms). So what do these disorders have in common? In each instance the disruptions lead to inefficiency in the system. CONCLUSION: Given this potential commonality, we propose that some of the methods used by behavioral researchers to study postural dynamics may also be of great utility to health care practitioners and psychologists alike. Furthermore we propose that this link will provide a framework that will allow scientists to address seemingly intractable problems such as motion sickness or subluxation. PMID- 11416826 TI - Rapid progression of acute sciatica to cauda equina syndrome. AB - OBJECTIVE: To demonstrate the importance of clinical examination and continued vigilance for neurologic deterioration in patients with sciatica. Cauda equina syndrome, a rare sequela of sciatica, is considered a medical emergency requiring surgical decompression. CLINICAL FEATURES: A 32-year-old woman had sciatica that rapidly progressed to cauda equina syndrome. Magnetic resonance imaging revealed the presence of a large nonsequestered disk fragment in the lower lumbar spine. INTERVENTION AND OUTCOME: The disk fragment was surgically excised. The patient experienced immediate pain relief after surgery but retained neurologic deficits. After 6 months of rehabilitation, neurologic integrity was restored, aside from patchy sensory loss of the left foot and buttocks. At the 6-month follow-up, the patient's sciatica had not returned. CONCLUSIONS: Most cases of sciatica, regardless of cause, will self-resolve; as a result, there might be a tendency to maintain a low index of suspicion for serious, progressive disorders such as cauda equina syndrome. Patients need to be educated as to signs of this emergency condition and informed as to the possible consequences of delaying treatment. By maintaining a high index of suspicion for any case that fails to respond as expected to a course of conservative therapy or that demonstrates signs of cauda equina syndrome, chiropractors can assume a pivotal role by investigating and referring appropriately and by aiding in active rehabilitation postoperatively. PMID- 11416827 TI - Posttraumatic osteolysis of the distal clavicle: analysis of 7 cases and a review of the literature. AB - OBJECTIVE: To discuss the clinical manifestation, radiographic features, and treatment of 7 cases of posttraumatic osteolysis of the distal clavicle. Also, to furnish evidence indicating that the current terminology for this disorder is ambiguous and to propose a new classification system. CLINICAL FEATURES: Three cases resulted from acute trauma, and 4 cases were caused by sports-related repetitive microtrauma. All the cases involved young male patients who had similar clinical presentations that included shoulder pain with decreased shoulder range of motion. Radiographic findings ranged from small erosive changes to aggressive osteolysis of the distal clavicle. INTERVENTION AND OUTCOME: All the patients were treated with conservative care consisting of sling immobilization, ice, various physiotherapeutic modalities, and mobilization exercises. The patients that followed through with care showed clinical improvement within 3 months. Follow-up radiographic examinations, when performed, demonstrated reconstitution of the distal clavicle of various degrees, although lagging behind clinical evidence of improvement. CONCLUSION: Traumatic osteolysis of the distal clavicle may result from acute trauma or repetitive microtrauma. Radiographic changes are varied, including irregularity of the distal clavicle, cystic erosions, and blatant osteolysis. Positive outcomes may be achieved with conservative care; however, patient compliance plays a fundamental role in the overall prognosis. PMID- 11416828 TI - The evidence in evidence-based practice: what counts and what doesn't count? PMID- 11416829 TI - Thoracic disk herniation: a case report. PMID- 11416831 TI - Saphenous vein harvesting by 'stripping' technique and 'W'-shaped patch covering after plaque incision in treatment of Peyronie's disease. AB - Harvesting of the saphenous vein tract by means of leg stripping is proposed in the treatment of Peyronie's disease. The technique of W-shaped saphenous vein after plaque incision to correct severe penile deformity associated with Peyronie's disease is described. Graft material was obtained from the lower saphenous vein by means of distal 'leg short stripping' technique. The size and number of tunical incisions depended on the size of the plaque. A 15 cm venous segment is generally sufficient to cover the defect. The venous segment used was W-shaped, assembled with 6/0 polydioxanone (PDS) uninterrupted sutures and then sutured to the albuginea defect. In our preliminary series of eight patients, penile shortening and erectile dysfunction is absent. Complete correction of penile deformity was achieved in seven patients (87.5%). One patient had minimal residual curvature (<20 degrees ) which did not result in difficulty with intromission. Saphenous harvesting by the stripping technique is not an invasive procedure and is quick and simple to perform. The W-shaped assembling technique is safe because the piece of saphena is kept intact and may be suited properly to the albuginea defect. PMID- 11416832 TI - Is there any association between Peyronie's disease and serum collagen markers? AB - Several theories regarding the pathogenesis of Peyronie's disease have been investigated under many clinical conditions. We have investigated the association of Peyronie's disease with the most common markers of collagen disease. Several serum markers of collagen disease (mucoproteins, C-reactive protein, antinuclear antibody, rheumatoid factor, lupus erythematosus cells, proteinograms) of 30 patients with Peyronie's disease were compared with those obtained from 30 patients, matched for age, with other urological conditions unrelated to the penis. Mucoproteins were altered in 66.7% of patients of the Peyronie's disease group and in 46.7% of the control patients (P>0.05). C-reactive protein was altered in 23.3% of the Peyronie's disease patients and in 13.3% of the control patients (P>0.05). Antinuclear antibody (ANA) was reactive in 16.7% of the tested group and in 6.7% of the control group (P>0.05). The rheumatoid factor was elevated in 6.7% of the patients from both groups (P>0.05). LE cells were normal in all the patients in our study. No statistical significance between the two groups was found in the protein electrophoresis test. Only the Waaler-Rose test (rheumatoid hemagglutination test) was statistically significant in our study (P<0.05). We have not found any significant association between the serum markers of collagen diseases in patients with Peyronie's disease, except the rheumatoid hemagglutination test (Waaler-Rose). PMID- 11416833 TI - Epidemiology of erectile dysfunction: results of the 'Cologne Male Survey'. AB - The last few decades have seen a marked increase in mean life expectancy in Central Europe. This has made elderly people and their quality of life a matter of ever-increasing medical concern. Available data from the United States and Scandinavia relating to erectile dysfunction (ED) do not enable us to draw valid conclusions about the current situation in Germany. The aim of the present study was to evaluate the epidemiology of male sexuality in Germany, and the proportion of men who need medical treatment because of increased suffering from this.A newly developed and validated questionnaire on male erectile dysfunction was mailed to a representative population sample of 8000 men, 30-80 y of age in the Cologne urban district. The response included 4489 evaluable replies (56.1%). The response rates in different age groups ranged from 49.2% to 68.4%. Regular sexual activity was reported by 96.0% (youngest age group) to 71.3% (oldest group). There were 31.5%-44% of responders who were dissatisfied with their current sex life. The prevalence of ED was 19.2%, with a steep age-related increase (2.3 53.4%) and a high co-morbidity of ED with hypertension, diabetes, pelvic surgery and 'lower urinary tract symptoms'. When treatment need was defined by co occurrence of ED and dissatisfaction with sex life, 6.9% men required treatment for ED. Oral treatment of ED was preferred by 73.8% of respondents. There were 46.2% respondents who were willing to contribute more than DM 50 (25 Euro) per month for ED treatment. We conclude that regular sexual activity is a normal finding in advanced age. ED is a frequent disorder, contributing to dissatisfaction with sex life in a considerable proportion of men. The high burden of ED is reflected in willingness to pay for treatment. ED is frequently associated with chronic diseases. Therefore adequate diagnostic workup is essential, to offer patients individually adapted treatment. General non reimbursability of treatment for ED appears to be unacceptable. PMID- 11416834 TI - Intracavernosal adrenalin injection in priapism. AB - Prolonged erection is a rare problem in urology but it must be treated effectively. The most common etiological factor is intracavernosal vasoactive agent injection for diagnosis or treatment of erectile dysfunction. The aim of this study was to evaluate the efficacy of intracavernosal adrenalin injection alone in the treatment of priapism. Nineteen patients with prolonged erection were evaluated. Seventeen out of the 19 prolonged erections were due to intracavernosal vasoactive agent injection and the remaining two were idiopathic. In all cases 2 ml adrenalin (1/100 000) was injected in each cavernosal body. In the patients who did not respond to the first injection, repeated adrenalin injections were performed at 20 min intervals. Blood pressure and heart rate were monitored during the injections. Detumescence was achieved in ten (53%) patients after the first injection. Repeated adrenalin injections (2-5 injections) were required in nine patients and eight (42%) of them achieved detumescence. Only one (5%) patient who had 26-h prolonged erection could not achieve detumescence. There was no significant difference in blood pressure and heart rate during the monitoring of the patients when compared to the initial values. No standard treatment method has yet been described for prolonged erection. Repeated aspirations and irrigations for treatment of prolonged erection are problematical applications both for patients and urologist. Intracavernosal adrenalin injection alone can be used with high efficacy and safety for the treatment of prolonged erection especially in patients with a short duration of erection. PMID- 11416835 TI - The mechanics of corporal veno-occlusion in penile erection: a theory on the effect of stretch-associated luminal constrictability on outflow resistance. AB - In order to further our understanding of the physiology of corporal veno occlusion, we developed a theory of a possible contribution to corporal venous outflow resistance which occurs as the result of venule stretching with resultant luminal narrowing when penile volume increases during the erection process. We stretched non-biological tubes and rabbit abdominal vena cava segments, performed flow-based and volume-based experiments to calculate the magnitude of N, the newly defined 'stretch-associated luminal constrictability' factor. We solved for (R(s)/R(u)), the ratio of the venule fluid resistance in the stretched state (R(s)) to the unstretched state (R(u)), to quantify the projected increases in fluid resistance as well as Q.R(u) where Q is the subtunical venule flow rate. For a given tube, N was found to be essentially constant for different amounts of stretch. A theory was formulated which predicted R(s) and Q as a function of N, DeltaP (intracavernosal pressure increase); V(E)/V(F) (tunical distensibility); X (cavernosal expandability) and R(u). Based on the magnitude of N=2, this theory predicts that patients with the highest values of both V(E)/V(F) and X would have maximal R(s) values, approaching infinity (complete occlusion) at a low DeltaP near 5 mmHg. In contrast, patients with low values of both V(E)/V(F) (eg Peyronie's disease) and X (eg corporal fibrosis), would be predicted to have minimal R(s) values. For example, a hypothetical patient with the lowest values of V(E)/V(F) and X would yield R(s) values only approaching 7.9 times that of unstretched values at a DeltaP increase of 90 mmHg. We concluded to that stretch associated venule resistance may occur as a result of decreased sub-tunical venule diameter and increased sub-tunical venule length. In individual patients, stretch-associated venule resistance may either dominate or be a minor component of the overall mechanism of corporal veno-occlusion. PMID- 11416836 TI - Predicting penile size during erection. AB - The aim of this prospective study was to identify clinical and engineering parameters of the flaccid penis for prediction of penile size during erection. Dorsal and ventral penile lengths, as well as base and tip circumferences were measured in flaccid states, gently stretched states and at full erection resulting from intracavernosal injection of prostaglandin E1 in 55 patients. The forces required to stretch the penis were measured by a specially designed gauge and regression relationships of the measured dimensions were calculated. An engineering model was developed to analyze differences between results obtained during stretching and erection, as well as to approximate the optimal force values which should be applied during the stretching part of the clinical evaluation of penile size. The ratio between the flaccid to stretched penile lengths was shown to be the best predictor for the ventral elongation from flaccid to erect penile lengths. The engineering analysis predicted that a minimal tension force of approximately 450 g during stretching of the penis is required to reach the potential erection length. The stretching forces exerted by the urologist in the clinical setting were experimentally shown to be significantly (P<0.01) less than this value. The values of the relative and absolute elongations of the stretched penis at its ventral aspect provide reliable estimations of its potential maximal elongation during erection. The model designed for this study may obviate the use of intracavernosal injections for estimating penile length during erection. PMID- 11416837 TI - Intracavernosal injections of vascular endothelial growth factor protects endothelial dependent corpora cavernosal smooth muscle relaxation in the hypercholesterolemic rabbit: a preliminary study. AB - Atherosclerosis is a major risk factor for erectile dysfunction, and loss of endothelium-dependent vasodilation appears early in the development of this disorder. Nitric oxide (NO) appears to be the principle mediator of erectile function and is generated in part by the sinusoidal endothelium. Vascular endothelial growth factor (VEGF) is an angiogenic growth factor and an endothelial cell-specific mitogen and the actions of VEGF are coupled to NO. In this preliminary study, we investigated whether VEGF could be used to protect endothelial dependent cavernosal relaxation from the atherosclerotic injury induced by a hypercholesterolemic diet.Two groups of New Zealand white adult male rabbits received a 1% cholesterol diet for four weeks, and two groups consumed normal rabbit chow. Half of the rabbits consuming the 1% cholesterol diet received weekly penile injections of 0.3 mg VEGF (n=8), and half injections of normal saline (n=8). Rabbits fed normal chow followed a similar protocol, half received weekly penile injections of 0.3 mg VEGF (n=6) and half were given weekly penile injections of normal saline (n=6). Isometric tension studies (with norepinephrine, acetylcholine, sodium nitroprusside and histamine) were performed on isolated strips of corpora cavernosa. The degree of corporal smooth muscle relaxation in response to ACH and SNP administration was recorded and compared. Significant elevation in serum total cholesterol levels occurred in rabbits receiving 4 weeks of the 1% cholesterol diet (727+/-75.6 mg/dl vs 38.7+/-5.53 mg/dl) P<0.01. There were no significant differences in cavernosal contraction in any group, while cavernosal smooth muscle from rabbits on normal chow retained the ability to relax in response to ACH and SNP in tissue bath. The hypercholesterolemic rabbits receiving VEGF had a significantly higher maximal per-cent relaxation to ACH (111+/-28.9) compared to the hypercholesterolemic rabbits that received NS (77+/-23.1, P<0.001). This difference in percent maximal relaxation to SNP was also present for hypercholesterolemic/VEGF rabbits (129.4+/ 24) versus the hypercholesterolemic/NS rabbits (115.0+/-18, P=0.033). In conclusion, intracavernosal injections of VEGF appear to protect corporal endothelium from hypercholesterolemia induced injury, thus preserving endothelial dependent corporal smooth muscle relaxation in hypercholesterolemic rabbit. PMID- 11416838 TI - Effects of castration and androgen replacement on erectile function in a rabbit model. PMID- 11416839 TI - The sexual health inventory for men (IIEF-5): reply to Vroege. PMID- 11416842 TI - Effect of plasma and matrix proteins on defensin-induced impairment of phagocytic killing by adherent neutrophils. AB - Infection is too often associated with prosthetic devices. Increased susceptibility to infection at these surgical sites appears to be associated with defective local phagocytic killing. The mechanisms for neutrophil down regulation, however, continue to be obscure. We have recently demonstrated that cytotoxic substances are released from granulocytes associated with materials. One group of releasants, the cationic human neutrophil peptide(s) (also called defensins) not only impairs the antimicrobial capacity of the granulocyte that releases it but also impairs bystander phagocytes. Because plasma or matrix proteins soon become associated with implants, we investigated the interactive effect of adding these proteins, singly and in combination, on the microbicidal effect of bystander cells. Some plasma/matrix proteins (whole plasma, albumin, fibrinogen, and fibronectin) strongly interfered with the anti-microbicidal effects generated by neutrophil-polystyrene interaction. Other proteins (vitronectin and laminin) were without effect. These results suggest that protein composition at the prosthetic implant site could have a significant effect on infectivity, depending on whether neutrophils releasants were attenuated. In the absence of attenuation, the local environment would be hostile to host defenses, permitting bacterial survival and proliferation. PMID- 11416843 TI - Poly(DL-lactic-co-glycolic acid) sponge hybridized with collagen microsponges and deposited apatite particulates. AB - A novel three-dimensional porous scaffold has been developed for bone tissue engineering by hybridizing synthetic poly(DL-lactic-co-glycolic acid) (PLGA), naturally derived collagen, and inorganic apatite. First, a porous PLGA sponge was prepared. Then, collagen microsponges were formed in the pores of the PLGA sponge. Finally, apatite particulates were deposited on the surfaces of the collagen microsponges in the pores of PLGA sponge. The PLGA-collagen sponge served as a template for apatite deposition, and the deposition was accomplished by alternate immersion of PLGA-collagen sponge in CaCl(2) and Na(2)HPO(4) aqueous solutions and centrifugation. The deposited particulates were small and scarce after one cycle of alternate immersion. Their number and size increased with the number of alternate immersion cycles. The surfaces of collagen microsponges were completely covered with apatite after three cycles of alternate immersion. The porosity of the hybrid sponge decreased gradually as the number of alternate immersion increased. Energy-dispersive spectroscopy analysis and X-ray diffraction spectra showed that the calcium-to-phosphorus molar ratio of the deposited particulates and the level of crystallinity increased with the number of alternate immersion cycles, and became almost the same as that of hydroxyapatite after four cycles of alternate immersion. The deposition process was controllable. Use of the PLGA sponge as a mechanical skeleton facilitated formation of the PLGA-collagen-apatite hybrid sponge into desired shapes and collagen microsponges facilitated the uniform deposition of apatite particulates throughout the sponge. The PLGA-collagen-apatite hybrid sponge would serve as a useful three-dimensional porous scaffold for bone tissue engineering. PMID- 11416844 TI - Control of shape and size of vascular smooth muscle cells in vitro by plasma lithography. AB - The ability to control the shape and size of cells is an important enabling technique for investigating influences of geometrical variables on cell physiology. Herein we present a micropatterning technique ("plasma lithography") that uses photolithography and plasma thin-film polymerization for the fabrication of cell culture substrates with a cell-adhesive pattern on a cell repellent (non-fouling) background. The micron-level pattern was designed to isolate individual vascular smooth muscle cells (SMC) on areas with a projected area of between 25 and 3600 microm(2) in order to later study their response to cytokine stimulation in dependence of the cell size and shape as an indication for the phenotypic state of the cells. Polyethylene terephthalate substrates were first coated with a non-fouling plasma polymer of tetraglyme (tetraethylene glycol dimethyl ether). In an organic lift-off process, we then fashioned square- and rectangular-shaped islands of a thin fluorocarbon plasma polymer film of approximately 12-nm thickness. Electron spectroscopy for chemical analysis and secondary ion mass spectroscopy were used to optimize the deposition conditions and characterize the resulting polymers. Secondary ion mass spectroscopy imaging was used to visualize the spatial distribution of the polymer components of the micropatterned surfaces. Rat vascular SMC were seeded onto the patterned substrates in serum-free medium to show that the substrates display the desired properties, and that cell shape can indeed be controlled. For long-term maintenance of these cells, the medium was augmented with 10% calf serum after 24 h in culture, and the medium was exchanged every 3 days. After 2 weeks, the cells were still confined to the areas of the adhesive pattern, and when one or more cells spanned more than one island, they did not attach to the intervening tetraethylene glycol dimethyl ether (tetraglyme) background. Spreading-restricted cells formed a well-ordered actin skeleton, which was most dense along the perimeter of the cells. The shape of the nucleus was also influenced by the pattern geometry. These properties make the patterned substrates suitable for investigating if the phenotypic reversion of SMC can be influenced by controlling the shape and size of SMC in vitro. PMID- 11416845 TI - Strain rate dependent orthotropic properties of pristine and impulsively loaded porcine temporomandibular joint disk. AB - The purpose of this study was to characterize the tensile stress-strain behavior of the porcine temporomandibular joint (TMJ) disk with respect to collagen orientation and strain rate dependency. The apparent elastic modulus, ultimate tensile strength, and strain at maximum stress were measured at three elongation rates (0.5, 50, and 500 mm/min) for dumbbell-shaped samples oriented along either anteroposterior or mediolateral axes of the disks. In order to study the effects of impact-induced fissuring on the mechanical behavior, the same properties were measured along each orientation at an elongation rate of 500 mm/min for disks subjected to impulsive loads of 0.5 N. s. The results suggested a strongly orthotropic nature to the healthy pristine disk. The values for the apparent modulus and ultimate strength were 10-fold higher along the anteroposterior axis (p < or = 0.01), which represented the primary orientation of the collagen fibers. Strain rate dependency was evident for loading along the anteroposterior axis but not along the mediolateral axis. No significant differences in any property were noted between pristine and impulsively loaded disks for either orientation (p > 0.05). The results demonstrated the importance of choosing an orthotropic model for the TMJ disk to conduct finite element modeling, to develop failure criteria, and to construct tissue-engineered replacements. Impact-induced fissuring requires further study to determine if the TMJ disk is orthotropic with respect to fatigue. PMID- 11416846 TI - Increase in free radicals on UHMWPE hip prostheses components due to inflamed synovial cell products. AB - Oxidative degradation of artificial UHMWPE joint implants caused by gamma-ray sterilization is thought to be responsible for the production of wear debris resulting in adverse tissue responses. On the other hand, it is well known that inflammation is associated with generation, by inflammatory cells, of free radicals (H(2)O(2) and NO) and destructive proteolytic enzymes (collagenases), which creates a strong oxidative environment. We hypothesized that when an UHMWPE implantation was performed in an inflammatory joint environment, the oxidative substances produced by inflamed synoviocytes could increase oxidative degradation of the polyethylene insert. We measured the amount of free radicals on conventional and on Duration-treated polyethylene samples by the electron spin resonance (ESR) technique before and after exposure of the samples to (1) inflamed synovial cell cultures; (2) normal synovial cell cultures; and (3) medium alone. We observed an increase in the number of free radicals on polyethylene samples after their immersion in cell cultures. Furthermore, it was observed that the increase of free radicals on polyethylene correlated with the degree of inflammation of synovial cells in culture. PMID- 11416847 TI - Influence of coating strain on calcium phosphate thin-film dissolution. AB - The success of calcium phosphate (CaP) coatings used to accelerate initial bone growth onto dental implants can vary depending on the CaP phases present in the coating. In this study, the effect of CaP coating crystal structure and morphology on dissolution rates was investigated. RF magnetron-sputtered CaP coatings (NTC) were compared to a less strained coating (HTC) obtained from heat treatment of sputtered samples at 550 degrees C. Coating strain differences were apparent in XRD spectra where hydroxyapatite-like planes shifted by 0.5 degrees 2theta and 0.05 degrees 2theta for the NTC and HTC coatings, respectively. HTC XRD peak widths were broader than NTC peak widths, indicating smaller crystals or grain sizes. These differences in grain size were corroborated by imaging with scanning probe microscopy. NTC coatings dissolved at a 300% faster rate than HTC coatings. A major factor contributing to this kinetic effect was the level of strain in both coatings. These results suggest an alternate design for CaP coatings can be obtained through the manipulation of coating strain. Using this approach, delivery of different ionic gradients from CaP coatings to surrounding tissue environments can be obtained from surfaces having similar chemistries. PMID- 11416848 TI - Mapping critical sites in collagen II for rational design of gene-engineered proteins for cell-supporting materials. AB - Collagen II is the most abundant protein of cartilage and forms a network of fibrils extended by proteoglycans that enables cartilage to resist pressure. The surface of the collagen fibril serves as a platform for the attachment of collagen IX, growth factors, and cells. In this study we examined the mechanism of the interaction of chondrocytes with recombinant versions of procollagen II, in which one of the four blocks of 234 amino acids that define repeating D periods of the collagen triple helix has been deleted. Analysis of the attachment of chondrocytes to collagen II variants with deleted D periods indicated that the collagen II monomer contains randomly distributed sites critical for cell binding. However, as was shown by spreading and migration assays, the D4 period, which is between residues 703 to 936, contains amino acids critical for cell motility. We also showed that binding, spreading, and migration of chondrocytes through three-dimensional nanofibrillar collagenous matrices are controlled by an interaction of the collagen triple helix with beta1 integrins. The results of this study provide a basis for the rational design of a scaffold containing genetically engineered collagen with a high density of specific sites of interaction. PMID- 11416849 TI - Erroneous aluminum and cobalt tissue concentrations from using formalin. AB - Because of the liberation of metal particles from orthopedic implants, concerns have been voiced about their long-term safety. To document the presence and measure the amount of metal in periprosthetic tissues, many reports have analyzed tissue specimens obtained from revision surgery. Some report the inclusion of formalin in tissue processing. However, the unavailability of any medical-grade formalin solution with a known metal content raises the suspicion of sample contamination. The purpose of this study was to measure the concentration of the metals in the formalin found in common orthopedic alloys and to assess this chemical's role in removing metal ions from tissue samples. To do this, tissues with known metal concentrations of cobalt and aluminum were mixed with formalin, and the differences in the formalin and tissue concentrations of these elements were evaluated. Tissue and formalin samples were tested prior to and immediately after their mixing and then at three 10-day intervals. At the end of the study additional unmixed samples of formalin and tissue for both metals were tested to ensure there had been no contamination. Both cobalt and aluminum were found in off-the-shelf formalin, with observed decreases in tissue metal concentration and increases in formalin metal concentration. PMID- 11416850 TI - In vitro biocompatibility study of electrically conductive polypyrrole-coated polyester fabrics. AB - This study investigated the basic biocompatibility aspects of two types of polypyrrole (PPy)-coated polyester fabrics for possible use as vascular prostheses. These PPy-coated fabrics, PPy-Phos and PPy-Plas, were sterilized with ethylene oxide (EO) and the following characterizations were performed: surface morphology by scanning electron microscope, EO residuals analysis by the headspace method, acute systemic toxicity in the mouse model, hemolysis, blood coagulation time, viability and proliferation of endothelial cells measured with the WST-1 method, and activation of polymorphonuclear (PMN) cells indicated by the specific expression of interleukin 8 mRNA measured by reverse transcription polymerase chain reaction. Virgin polyester fabrics, expanded poly(tetrafluoroethylene) (ePTFE), and medical-grade Bionate 80A poly(carbonate urethane) were used as references in the cell culture experiments. The PPy-coated fabrics revealed different surface morphologies by showing more PPy lamina and clusters on the PPy-Plas. Neither of the PPy-coated fabrics had an adverse effect on hemolysis and coagulation time, and they did not cause any acute systemic toxicity. The EO residual level was as low as 5 ppm or less, which is considered quite acceptable. Although exhibiting a relatively low initial cell adhesion at 24 h, the two PPy-coated samples showed no cytotoxicity at 72 and 168 h. Bionate 80A and ePTFE recorded cytotoxicity at 72 and 168 h, respectively. The virgin fabrics also demonstrated a decrease of viable cells at 72 h that was not significant. The activation of PMN cells induced by both PPy-coated fabrics, the ePTFE, and the negative control was significantly lower than that induced by their respective tumor necrosis factor-alpha controls. These results therefore highlighted the potential of PPy-coated fabrics for use as cardiovascular prostheses. It was suggested that cell adhesion moieties should be incorporated into the PPy/fabric composite to increase cell adhesion and subsequent cell proliferation. PMID- 11416851 TI - Preservation of platelet function on 2-methacryloyloxyethyl phosphorylcholine graft polymer as compared to various water-soluble graft polymers. AB - The chemical structures of water-soluble polymers grafted onto PE surfaces affect platelet function when the platelets contact the polymer surfaces. To improve our understanding of this effect, this study sought to control the blood/materials interaction on the surfaces of polyethylene (PE) by grafting with various water soluble polymers. Such polymers as poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC), poly(acrylamide) (PAAm), poly(N-vinylpyrrolidone) (PVPy), and poly[monomethacryloyl poly(ethylene glycol)] (PMPEG) were grafted on low-density PE sheets by photoinduced graft polymerization. Both the PE bags modified with water-soluble polymers and those nonmodified were prepared by heat processing. Activation of platelets after storage in the PE bags was evaluated by measuring the cytoplasmic free calcium ion concentration ([Ca(2+)]i). The concentration of [Ca(2+)]i of platelets in contact with the PE surface grafted with PMPC was the same as that of native platelets and significantly less than that in contact with other PE surfaces grafted with water-soluble polymers. The number of adherent platelets was effectively decreased on PE surfaces grafted with PMPC and PMPEG, as compared with nontreated PE. The aggregation ability of platelets was also measured after storage of platelet-rich plasma in the PE bags. The PE surface grafted with PMPC effectively maintained aggregation ability as compared with both the nontreated PE and with PE grafted with PAAm, PVPy, and PMPEG. It was concluded that for preserving platelet function, PMPC was the most effective of these water-soluble polymers used for surface modification. PMID- 11416852 TI - Improvement of swelling properties of poly(2-hydroxyethyl methacrylate) hydrogel by means of biomimetic method. AB - Poly(2-hydroxyethyl methacrylates) (PHEMAs) structurally modified by means of polymer blends and random copolymers are intensively studied in order to improve mechanical properties. It was recently shown that a hydroxyapatite coating, which should improve the bonding of this biomaterial to the bone, can be obtained by means of the biomimetic method. When PHEMA is submitted to the biomimetic method, its swelling ratio is improved. This can be ascribed to the deposition of a silicatic layer, which improves the hydrophilicity, on the surface of the internal pores during the first stage of the method. This appears to be a valuable result for producing modified PHEMAs with improved mechanical properties and good swelling. The experimental results indicate the following: stronger interactions with the water molecules are set up, an induction period is observed that is linked to the rate of the reactions occurring at the surface of the glass and the establishment of a convenient concentration of the silicate ions at the external surface of the polymer, and the diffusion of simulated body fluid into the pores is the limiting stage of the process. PMID- 11416853 TI - Integrin expression at the bone/biomaterial interface. AB - The aim of this study was to visualize integrin expression by cells in interface tissue in relation to their ligands. Tissue samples were obtained from 25 patients undergoing revision of aseptically loose total joint replacements. Serial sections were immunolabeled for the integrins alpha(2)beta(1), alpha(v)beta(3), alpha(4)beta(1), alpha(L)beta(2) (CD11a), alpha(M)beta(2) (CD11b), and alpha(X)beta(2) (CD11c), and the ligands fibronectin, laminin, vitronectin, intercellular adhesion molecule-1, and vascular adhesion molecule-1. Most cells were found to express alpha(2)beta(1), most macrophages and giant cells expressed CD11b, and the majority of CD11a was found on perivascular T lymphocytes. From the small amount of alpha(4)beta(1) and vascular adhesion molecule-1 expression in the interface tissue and the combination of CD11a, CD11b, and intercellular adhesion molecule-1 expression, it would seem that macrophages use beta(2) integrins to transmigrate. PMID- 11416854 TI - Effect of molecular weight of thermoreversible polymer on in vivo retention of rhBMP-2. AB - To investigate the effect of polymer molecular weight (MW) on rhBMP-2 delivery by thermoreversible polymers, four polymers with similar lower critical solution temperatures (20 degrees -22 degrees C) but different MWs were studied. Thermoreversible polymers were based on N-isopropylacrylamide (NiPAM), ethyl methacrylate (EMA), and N-acryloxysuccinimide (NASI), and had MWs of either approximately 49 kDa or approximately 400 kDa. The NASI content was either 0 or 1 1.6%. High MW polymers, irrespective of their NASI content, formed a stable gel with significantly lower water uptake and exhibited a dense micelle with average pore size smaller than the low MW polymers. NiPAM/EMA polymers without NASI did not conjugate with recombinant human bone morphogenetic protein-2 (rhBMP-2). NiPAM/EMA polymers containing NASI, however, gave conjugation with rhBMP-2. For polymers without NASI, a high MW was essential for rhBMP-2 retention when injected intramuscularly in Sprague-Dawley rats. For NASI-containing polymers, the MW of the polymer did not make a significant difference because rhBMP-2 retention was equivalent for different size polymers. We conclude that polymer MW affects rhBMP-2 retention in vivo in polymers designed for physical entrapment of rhBMP-2, but not in polymers designed for chemical conjugation with rhBMP-2. PMID- 11416855 TI - Ultrastructure of the interface between bioactive composite and bone: comparison of apatite and wollastonite containing glass-ceramic filler with hydroxyapatite and beta-tricalcium phosphate fillers. AB - We have developed a bioactive bone cement that consists of apatite and wollastonite containing glass-ceramic (AW-GC) powder and bisphenol-a-glycidyl dimethacrylate (Bis-GMA)-based resin. In this study, we made three types of composite (designated AWC, HAC, and TCPC) consisting of AW-GC, hydroxyapatite (HA,) or beta-tricalcium phosphate (beta-TCP) powder as the inorganic filler and Bis-GMA-based resin as the organic matrix. The proportion by weight of the filler mixed into the cement was 70%. Rectangular plates (10 x 15 x 2 mm) of each composite were made and abraded with 2000 alumina powder. These composites were implanted into tibial metaphyses of rabbits. Specimens were prepared 10 and 25 weeks after implantation and examined using transmission electron microscopy (TEM). AWC was in direct contact with bone 10 weeks after implantation, and AW-GC particles were partially absorbed at the surface. HAC was in contact with partially mineralized extracellular matrix 10 weeks after implantation. In TCPC implanted specimens, randomly oriented mineral was observed 10 weeks after implantation; however, collagenous extracellular matrix rarely was observed. In 25-week specimens, AW-GC particles were completely absorbed and replaced by new bone, and there was no intervening soft tissue. Both HAC and TCPC were in contact with bone at 25 weeks. These results indicate that AWC has higher bioactivity than either HAC or TCPC. PMID- 11416856 TI - Effects of anisotropy on elastic moduli measured by nanoindentation in human tibial cortical bone. AB - Many biological materials are known to be anisotropic. In particular, microstructural components of biological materials may grow in a preferred direction, giving rise to anisotropy in the microstructure. Nanoindentation has been shown to be an effective technique for determining the mechanical properties of microstructures as small as a few microns. However, the effects of anisotropy on the properties measured by nanoindentation have not been fully addressed. This study presents a method to account for the effects of anisotropy on elastic properties measured by nanoindentation. This method is used to correlate elastic properties determined from earlier nanoindentation experiments and from earlier ultrasonic velocity measurements in human tibial cortical bone. Also presented is a procedure to determine anisotropic elastic moduli from indentation measurements in multiple directions. PMID- 11416857 TI - Mechanical and rheological improvement of a calcium phosphate cement by the addition of a polymeric drug. AB - A polymeric acrylic system supporting a derivative of the aminosalicylic acid was incorporated in a calcium phosphate cement, with the aim not only to achieve some pharmacological effects but to obtain an improvement of its mechanical and rheological properties. It is known that, besides the analgesic and anti inflammatory properties, the salicylic group presents a calcium complexation ability. The inorganic phase of the cement consisted of alpha-tricalcium phosphate [alpha-Ca(3)(PO(4))(2)] and precipitated hydroxyapatite added as a seed. The liquid phase was an aqueous solution of Na(2)HPO(4). The polymeric drug increased the injectability of the cement. The hydrolysis of the alpha-tricalcium phosphate into calcium-deficient hydroxyapatite proceeded at a lower rate because of the addition of the polymeric drug. As a consequence, the cement hardening was slightly slower, although the final compressive strength was 25% higher. The bending strength increased from 5 to 9 MPa with the addition of the polymeric drug. The strengthening of the structure was related to the reduction of porosity and the lower size of the precipitated crystals, as observed by scanning electron microscopy. PMID- 11416858 TI - Apatite nucleation on silica surface: a zeta potential approach. AB - Zeta potential measurements on pure silica, prepared by the sol-gel method from tetraethoxysilane under acidic conditions, are reported in different suspensions. Water suspensions and suspensions containing calcium or phosphate ions with and without NaCl were tested. zeta potential measurements were carried out as a function of the pH and ion concentration. Also, calcium and phosphate adsorption on silica was determined experimentally. The results of zeta potential and adsorption measurements suggest that both calcium and phosphate ions can be adsorbed on the silica surface; however, calcium adsorption is stronger than phosphate adsorption. When calcium and sodium ions are present in the suspension, calcium adsorption decreases. It seems that certain sites on the silica surface are specific for calcium adsorption. PMID- 11416859 TI - Composites made of rapidly resorbable ceramics and poly(lactide) show adequate mechanical properties for use as bone substitute materials. AB - Porous composite materials made of poly(L, DL-lactide) and a ceramic component, alpha-tricalcium phosphate (alpha-TCP) or one of the rapidly resorbable glass ceramics, GB9N or GB14N, respectively, were developed to be used as bone substitutes. The present article describes the mechanical properties and the in vitro degradation characteristic of the different composite materials. The yield strength, the elastic modulus, and the molecular weight were measured after in vitro degradation up to 78 weeks. The initial strengths of the alpha-TCP composite (12.5 +/- 0.7 MPa) was higher than that of the GB9N and GB14N composites (8.3 +/- 0.2 MPa and 10.9 +/- 0.2 MPa, respectively). The initial elastic moduli of the composites were between 450 and 650 MPa. The mechanical properties remained constant until a degradation period of 26 weeks. Then they decreased continuously until they were completely lost at week 52. The molecular weight (M(w)) decreased steadily from 91,000 D in the case of the alpha-TCP composite and 78,000 D and 85,000 D in the case of the GB9N or GB14N composites, respectively, to about 10,000 D at week 78. It was concluded that the composites show adequate mechanical properties in the range of cancellous bone and a suitable degradation characteristic to be used as bone substitute materials. PMID- 11416860 TI - Proliferation and differentiation rates of a human osteoblast-like cell line (SaOS-2) in contact with different bone substitute materials. AB - The aim of our study was to investigate the influence of four bone substitutes on the growth behavior of a human osteoblast-like cell line (SaOS-2) culture: pure alpha tricalcium phosphate (alpha-TCP = BIOBASE), a bioactive glass (bioglass), a neutralized glass-ceramic (GB9N), and solvent dehydrated bone. We established an in vitro cell culture model with three-dimensional scaffolds (cubes of 0.7 x 0.7 x 1.0 cm) of porous bone substitutes to investigate proliferation and differentiation rates of SaOS-2 cells. The cultures were analyzed for individual cell morphology after 5 days of growing using scanning electron microscopy. Fracture preparations of the cubes showed that cells could infiltrate the porous structures, but the cell shapes varied from individual round-shaped cells to wide spread cells and cell clusters, depending on the material. Also, the differentiation of the seeded cells was dissimilar after a 5-day incubation. The specific alkaline phosphatase (ALP) enzyme activity (ALP/DNA) measured in the supernatants of alpha-TCP-grown cells was nine times higher than the lowest activity, as observed by cells incubated on GB9N. Early (Collagen1, ALP) and late marker (osteocalcin, bone sialoprotein) of osteoblastic differentiation were proofed by reverse transcriptase-polymerase chain reaction analysis. Cells grown on bone substitutes and bioglass seem to be less differentiated than alpha-TCP grown cells, because of noticeably less amounts of osteocalcin and bone sialoprotein. The cultivation on GB9N seems to dedifferentiate the cells, because even the ALP expression was reduced as well. Our results indicate that distinct bone substitutes influence proliferation and differentiation of osteoblastic cells in different manners. These results might influence the selection of an adequate bone substitute for clinical use as well, part from degradative and biomechanical properties. PMID- 11416861 TI - Formation of bone-like apatite on poly(L-lactic acid) fibers by a biomimetic process. AB - Bone-like apatite coating on poly(L-lactic acid) (PLLA) fibers was formed by immersing the fibers in a modified simulated body fluid (SBF) at 37 degrees C and pH 7.3 after hydrolysis of the fibers in water. The ion concentrations in SBF were nearly 1.5 times of those in the human blood plasma. The apatite was characterized by scanning electron microscopy (SEM) with energy dispersive X-ray spectroscopy (EDX), X-ray photoelectron spectroscopy (XPS), thin-film X-ray diffraction, and Fourier transform infrared spectroscopy. After 15 days of incubation in SBF, an apatite layer with about 5-6 microm thickness was formed on the surface of the fibers. This apatite had a Ca/P ratio similar to that of natural bone. The mass of apatite coated PLLA fibers increased with extending the incubation time. After 20 days incubation, the fibers increased their mass by 25.8 +/- 2.1%. The apatite coating had no significant effect on the tensile properties of PLLA fibers. In this article, the bone-like apatite coating on three-dimensional PLLA braids was also studied. The motivation for this apatite coating was that it might demonstrate enhanced osteoconductivity in the future studies when they serve as biodegradable scaffolds in tissue engineering. PMID- 11416862 TI - DNA methylation variation in cloned mice. AB - Mammalian cloning has been accomplished in several mammalian species by nuclear transfer. However, the production rate of cloned animals is quite low, and many cloned offspring die or show abnormal symptoms. A possible cause of the low success rate of cloning and abnormal symptoms in many cloned animals is the incomplete reestablishment of DNA methylation after nuclear transfer. We first analyzed tissue-specific methylation patterns in the placenta, skin, and kidney of normal B6D2F1 mice. There were seven spots/CpG islands (0.5% of the total CpG islands detected) methylated differently in the three different tissues examined. In the placenta and skin of two cloned fetuses, a total of four CpG islands were aberrantly methylated or unmethylated. Interestingly, three of these four loci corresponded to the tissue-specific loci in the normal control fetuses. The extent of aberrant methylation of genomic DNA varied between the cloned animals. In cloned animals, aberrant methylation occurred mainly at tissue-specific methylated loci. Individual cloned animals have different methylation aberrations. In other words, cloned animals are by no means perfect copies of the original animals as far as the methylation status of genomic DNA is concerned. PMID- 11416863 TI - Expression of thyroid transcription factor-1 (TTF-1) gene in the ventral forebrain and endostyle of the agnathan vertebrate, Lampetra japonica. AB - The Thyroid transcription factor-1 (TTF-1) gene belongs to the Nkx-2.1 subfamily and encodes a transcription factor containing an NK-2-type homeodomain. In our study, we isolated and characterized cDNA clones for the TTF-1/Nkx-2.1 orthologue (LjTTF-1) from the agnathan vertebrate Lampetra japonica. Spatial and temporal expression patterns assessed by in situ hybridization revealed the expression of LjTTF-1 in the anterior nerve cord and anteroventral region of the pharynx. The neural expression was subsequently restricted to the ventral diencephalon. The pharyngeal expression, on the other hand, extended posteriorly to the fourth pharyngeal-pouch level and was finally localized in the endostyle anlage. In the differentiated endostyle of ammocoete larvae, the expression of LjTTF-1 was chiefly detected in type 2a, 2b, and 2c cells, which develop adjacent to glandular cells. These expression patterns of LjTTF-1 support the idea that this gene family plays an important role in the development of the rostral brain and endostyle equivalent organs. Furthermore, histological comparisons between TTF 1/Nkx-2.1 expression in the endostyles of ammocoetes and ascidians suggested the possibility that the organogenetic architecture of the endostyle is conserved among chordates. PMID- 11416864 TI - Developmental regulation of an instability element from the Drosophila fushi tarazu mRNA. AB - The Drosophila fushi tarazu (ftz) mRNA is one of the shortest-lived metazoan mRNAs, and its instability is crucial for proper development of the embryo. Previously, we identified two cis-acting elements that are required for ftz mRNA degradation, one within the 5' one-third and another in the 3'UTR of the message. Here we focus on the 3'UTR element termed FIE3 (ftz instability element in the 3'UTR). To investigate the developmental regulation of the FIE3-dependent degrading activity we measured the abundance of an FIE3-containing mRNA in ovaries, unfertilized eggs, and different larval and adult tissues. We found that FIE3-degrading activity is present at all developmental stages and tissues examined, except in the ovary. Activation of the FIE3-dependent mRNA decay is independent of fertilization because it could be triggered by egg activation. Finally, we provide evidence that mutation of conserved elements within FIE3 had no effect on mRNA instability. PMID- 11416865 TI - Hypomorphic phenotype in mice with pituitary-specific knockout of steroidogenic factor 1. AB - The bacteriophage Cre recombinase provides a powerful approach for tissue specific gene inactivation. Using a Cre transgene driven by the common alpha subunit of glycoprotein hormones (alphaGSU-Cre), we have previously inactivated steroidogenic factor 1 (SF-1) in the anterior pituitary, causing hypogonadotropic hypogonadism with sexual infantilism, sterility, and severe gonadal hypoplasia. We now explore the molecular mechanisms underlying a hypomorphic gonadal phenotype in mice carrying two floxed SF-1 alleles (F/F) relative to mice carrying one recombined and one floxed allele (F/R). Because their Cre-mediated disruption of the locus encoding SF-1 was less efficient, alphaGSU-Cre, F/F mice retained some gonadotropin-expressing cells in the anterior pituitary, thereby stimulating some gonadal function. This novel in vivo model for exploring the effects of differing levels of gonadotropins on gonadal development highlights the need for careful genotype-phenotype comparisons in studies using Cre recombinase to produce tissue-specific knockouts. PMID- 11416866 TI - In utero complementation of a neural crest-derived melanocyte defect using cell directed gene transfer. AB - This study describes an in utero approach for overexpressing genes in a cell-type directed manner. It uses an avian leukosis retroviral expression system coupled with a transgenic mouse line expressing the viral receptor tv-a from a tissue specific promoter (RCAS-TVA system) (Federspiel et al., 1994, and reviewed in Fisher et al., 1999). A transgenic mouse line was generated expressing tv-a from the Dopachrome tautomerase promoter (DCT-tv-a) in embryonic melanocyte precursors (melanoblasts). RCAS virus encoding beta-galactosidase (RCAS-LacZ) or tyrosinase (RCAS-Tyr) was injected in utero into embryonic day 12.5 albino (tyrosinase inactive) mouse embryos. Animals were analyzed for beta-galactosidase activity or tyrosinase activity (hair pigmentation). RCAS gene expression was detected in 44% and 25% of the transgenic mice, respectively. We demonstrate the RCAS-TVA system coupled with the DCT-tv-a line of mice can be used for in utero infection. PMID- 11416867 TI - Utero-tubal transfer of mouse embryos. AB - A successful embryo transfer depends on the quality of the transferred embryos, recipients, and the transfer techniques. Among these, transfer techniques are often the limiting factor because transfer methodologies and personal skills vary. Suboptimal embryo transfer procedures can compromise transgenic experiments (pronuclear microinjection and gene targeting) and critical steps of mouse colony maintenance (embryo cryopreservation and mouse line rederivation). Here we present an efficient and simple procedure utilizing specific designs to improve the transfer quality. A 100% implantation rate is observed after the utero-tubal embryo transfer, which indicates that the modified method successfully prevents the embryos from flowing out of the punctured hole during embryo transfer. We believe this alternative methodology is able to fulfill the need of high efficiency of animal production. PMID- 11416868 TI - Transposition and gene disruption in the male germline of the mouse. AB - We have tested a synthetic, functional, transposon called Sleeping Beauty for use in mice as a germline insertional mutagen. We describe experiments in which mutagenic, polyadenylation-site trapping, transposon vectors were introduced into the germline of mice. When doubly transgenic males, expressing the Sleeping Beauty transposase gene (SB10) and harboring poly(A)-trap transposon vectors, were outcrossed to wild-type females, offspring were generated with new transposon insertions. The frequency of new transposon insertion is roughly two per male gamete. These new insertions can be passed through the germline to the next generation and can insert into or near genes. We have generated a preliminary library of 24 mice harboring 56 novel insertion sites, including one insertion into a gene represented in the EST database and one in the promoter of the galactokinase (Gck) gene. This technique has promise as a new strategy for forward genetic screens in the mouse or functional genomics. PMID- 11416869 TI - Effects of dexamethasone on proliferation, chemotaxis, collagen I, and fibronectin-metabolism of human fetal lung fibroblasts. AB - Premature infants at risk for bronchopulmonary dysplasia (BPD) are often treated with dexamethasone (Dex), which has been shown to suppress inflammatory processes in the lung. To elucidate a possible direct influence on the fibroproliferative component of the disease, we studied the effects of Dex in therapeutic and supratherapeutic dosages (5-50 nmol/L) on proliferation, chemotaxis, procollagen I, and fibronectin metabolism of human fetal lung fibroblasts in vitro. Proliferation was inhibited by Dex in a dose-dependent manner. Chemotactic activity in response to conditioned medium of human fetal fibroblasts also showed a dose-dependent inhibition after pretreatment with Dex. The amount of procollagen I C-terminal propeptide and fibronectin per cell in the cell culture supernatant was increased in the presence of Dex. Our results show that Dex does not uniformly suppress the fibroproliferative activity of human fetal lung fibroblasts, which may explain in part the unsatisfactory long-term effects of Dex treatment in BPD. PMID- 11416870 TI - Sex differences in response to steroids in preterm sheep lungs are not explained by glucocorticoid receptor number or binding affinity. AB - We recently reported that prenatal glucocorticoid therapy is less effective at promoting an improvement in lung function in male than in female sheep. This observation, and the higher incidence of respiratory distress syndrome in human males, suggests that the male fetal lung may be less responsive to glucocorticoids than is the female fetal lung. Since glucocorticoids are known to exert their effects via specific cytoplasmic glucocorticoid receptors (GR), we hypothesized that there may be sexual dimorphism in either the number or binding affinity of lung GR. To test the hypothesis, binding of dexamethasone (a synthetic glucocorticoid, 0.5-40 nM) by cytosolic fractions of male (n = 16) and female (n = 16) fetal sheep lung was measured at 125 days gestation (term = 148 days). Scatchard analysis of dexamethasone binding showed that the total number of GR (Bmax) did not significantly differ between male (346 +/- 42 fmol/mg protein) and female (277 +/- 23 fmol/mg protein) fetuses. The measured binding affinity (Kd) in male fetal lungs (6.85 +/- 0.43 nM) was not significantly different from that in females (8.46 +/- 1.02 nM). In conclusion, this study suggests that sex differences in fetal sheep lung responses to glucocorticoid therapy are not due to differences in the number or binding affinity of lung GR. PMID- 11416871 TI - Changes in arterial oxygen tension when weaning neonates from inhaled nitric oxide. AB - We set out to evaluate changes in arterial oxygen tension (PaO(2)) when weaning neonates from inhaled nitric oxide (INO). We reviewed the records of 505 prospectively collected INO weaning attempts on 84 neonates with hypoxic respiratory failure. PaO(2) values before and 30 min after weaning attempts were recorded. Relationships between change in PaO(2) and decreases in INO concentrations were investigated using regression analysis and ANOVA. PaO(2) decreased (-18.7 +/- 1.8 torr; P < 0.001); when weaning INO. A stepwise decline in PaO(2) was observed weaning INO from 40 ppm. The greatest decline occurred when INO was discontinued (-42.1 +/- 4.1 torr). Forward stepwise multiple regression using variables with significant relationships to the decline in PaO(2) identified the specific dose reduction 7(P < 0.001), the prewean PaO(2) (P < 0.001), and surfactant therapy (P = 0.018) as the variables best describing the change in PaO(2)(P = 0.004, r = 0.51). In conclusion, a graded decline in PaO(2) occurs when reducing INO. INO should be weaned to less than 1 ppm before discontinuing its use. Prior surfactant treatment appears to enhance the oxygenation reserve when weaning INO. PMID- 11416872 TI - Respiratory syncytial virus immune globulin: decisions and costs. AB - A decision analysis was used to evaluate the economic effectiveness of respiratory syncytial virus immune globulin (RSVIG) prophylaxis on selected pediatric populations at risk for developing RSV bronchiolitis or all respiratory illness-related hospitalizations. We compared costs, outcomes, and cost effectiveness of administering RSVIG to no treatment in different pediatric populations, including those at risk of developing RSV-bronchiolitis and those at risk of developing any respiratory illness-related hospitalization. We observed that if only infants at high risk of severe RSV infections received treatment with RSVIG, a calculated cost saving of about 27,000 dollars per hospitalization prevented were realized. If the Food and Drug Administration (FDA)-approved indications for RSVIG were followed, the cost to prevent one hospitalization due to RSV bronchiolitis would be over 53,000 dollars. If the aim, however, was to prevent all respiratory illness-related hospitalizations for this broader population, a much lower cost (4,000 dollars) to prevent one hospitalization would result. In this situation, cost neutrality was possible, with a therapy cost of 2,843 dollars compared to the actual average therapy cost of 4,444 dollars. Sensitivity analysis showed that the model was relatively insensitive to all variables, with the exceptions of costs related to RSVIG and intensive care unit (ICU) admissions. We conclude that RSVIG resulted in cost savings if therapy were reserved for the infants who are at highest risk for developing severe RSV infections. RSVIG is not cost-effective for preventing RSV bronchiolitis when used according to the FDA-approved indications. Education that emphasizes frequent hand-washing, avoidance of passive smoking, and lessening exposure to sick children remains the least expensive prevention tool. PMID- 11416873 TI - Effect of clarithromycin on airway obstruction and inflammatory markers in induced sputum in cystic fibrosis: a pilot study. AB - To determine whether macrolide antibiotics improve pulmonary function and decrease airway inflammation in cystic fibrosis (CF), we treated 10 patients (females; aged 19-26 years, all colonized with P. aeruginosa, none with atypical Mycobacteria) with 3 weeks of placebo, followed by 6 weeks of clarithromycin (500 mg BID) in a single-blind prospective study. We also determined the safety of sputum induction and the reproducibility of assessing inflammatory markers in induced sputum. Subjects performed spirometry and underwent sputum induction (12 min inhalation of 3% saline) at 3-week intervals. We found that sputum induction was well-tolerated. We also found that the reproducibility was high for neutrophil (PMN) number (R = 0.87, P = 0.009), interleukin (IL)-8 (R = 0.73, P < 0.05, free neutrophil elastase (NE) (R = 0.82, P < 0.05), and myeloperoxidase (MPO) levels (R = 0.86, P < 0.05), but was less so for tumor necrosis factor (TNF)-alpha (R = -0.15, P = 0.7). We found no significant difference in pulmonary function after 6 weeks of treatment with clarithromycin (FEV(1) (% predicted) (mean +/- SEM), 2.2 +/- 0.9 (60 +/- 24%) vs. 2.3 +/- 1 (61 +/- 29%)), and no significant differences in any of the inflammatory indices measured. The median (and range) values before and after treatment for indices of airway inflammation in the induced sputum samples were: for PMNs, 8 (1-326) and 21 (0.2 -175) x 10(6) cells/mL sputum; for IL-8, 156 (24-656) and 202 (16-680) ng/mL; for free NE, 260 (31-1,264) and 237 (49-1,048) microg/mL; for TNF-alpha, 20 (7-128) and 35 (17-87) pg/mL; and for MPO, 169 (13-960) and 195 (14-816) microg/mL. We conclude that clarithromycin is not uniformly effective in improving airway obstruction or in decreasing airway inflammation in patients with CF. PMID- 11416874 TI - Effect of intravenous antibiotics on exercise tolerance (3-min step test)) in cystic fibrosis. AB - Most children with cystic fibrosis (CF) feel better and display more energy after a course of intravenous antibiotics (IVABs), but this is not always reflected by a satisfactory improvement in lung function. We assessed the change in exercise tolerance after treatment with IVABs using the 3-min step test, and compared it with changes in spirometric lung function and arterial oxygen saturation (SaO(2)). Thirty-six children (mean age, 13.8 years) were enrolled from two tertiary CF centers during an inpatient stay for IVABs. After 10-14 days of treatment, there was a significant improvement in median FEV(1) from 43% to 57% of predicted values (P < 0.0001), and median FVC from 66% to 73% of predicted values (P < 0.0001), while median SaO(2) significantly increased from 95% to 96.5% (P < 0.05). This was accompanied by a reduction in resting heart rate (median 118 bpm to 109 bpm, P < 0.005) and subjective breathlessness at rest (median visual analogue score 2.2 to 0.8, P < 0.005). All outcomes of exercise tolerance were improved after IVABs. There was a reduction in maximum heart rate (median 156 bpm to 150 bpm, P < 0.05) and an increase in minimum SaO(2) (median 93.5% to 94.5%, P = 0.08) measured during the step test. There was also a reduction in subjective breathlessness (median visual analogue score of 5.5 to 4.2, P < 0.005) and objective breathlessness (median 15-count score of 3 to 2, P < 0.0001) measured immediately after the step test. Exercise testing was a useful outcome measure for monitoring effectiveness of inpatient therapy, and complemented spirometry and SaO(2) monitoring. The simple ward-based 3-min step test was found to be a particularly suitable method for measuring changes in exercise tolerance in children with CF. PMID- 11416875 TI - Inspiratory resistive load detection in children with life-threatening asthma. AB - The detection of inspiratory resistive (R) loads was studied in nonasthmatic children (NA), asthmatic children (A), and children with a history of life threatening asthma (LTA). It was hypothesized that the LTA children would have a reduced ability to detect added mechanical loads as measured by the Weber fraction, which assesses the resistive load detection threshold (DeltaR(50)/R(0)). Subjects were separated from the investigator, were seated in a soundproofed room, and breathed through a nonrebreathing valve with the inspiratory port connected to the loading manifold. The subject's inspiratory baseline resistance (R(aw)) was measured by the interrupter method. Ten magnitudes of R loads and no-load were presented randomly 10 times each for a single inspiration. The loads were presented in three trials. Subjects pressed a button if they detected the presence of a load. The DeltaR(50) was determined from the % detection-DeltaR curve. R(0) was the sum of the subject's R(aw) and the minimal resistance of the apparatus. The DeltaR(50)/R(0) for children with life- threatening asthma was significantly greater than for asthmatic and nonasthmatic children. The increased DeltaR(50)/R(0) suggests that children with LTA are at risk of life-threatening asthma attacks, in part because it requires a greater change in resistance above their baseline resistance before they sense an increased mechanical load such as presented to them by bronchoconstriction during an asthmatic attack. PMID- 11416876 TI - Efficacy of sublingual immunotherapy in children with asthma and rhinitis: a double-blind, placebo-controlled study. AB - To evaluate the efficacy of specific sublingual immunotherapy (SLIT), we enrolled 15 children with asthma and rhinitis (7 girls, 8 boys, mean +/- SD age of 11.7 +/ 3.3) allergic to house dust mite (HDM) into a double-blind, placebo-controlled study. After a run-in period, patients were randomized to receive either placebo (n = 7) or SLIT (n = 8) with a standardized Dermatophagoides pteronyssinus (D. pteronyssinus) + Dermatophagoides farinea (D. farinea) 50/50 extract. They received increasing doses up to 100 index units of reactivity (IR) every day for 4 weeks, then 100 IR/day for another 4 weeks, followed by maintenance therapy consisting of 20 drops 2 times a week for 4 months. Efficacy was assessed at the end of 6 months of therapy according to symptom and medication scores, serum total IgE levels, results of lung function tests, methacholine provocation tests, and skin prick tests. Daily means for the asthma score and use of inhaled beta-2 mimetics decreased significantly in the SLIT group (P = 0.05, P = 0.028, respectively), whereas no such difference was observed in the placebo group. At the end of follow-up, mean daily doses of intranasal steroids needed for control of rhinitis symptoms decreased significantly in the SLIT group (P = 0.04). Baseline skin sensitivity to D. pteronyssinus and D. farinea was not significantly different between in the two groups, whereas end-point wheal diameter obtained with D. pteronyssinus extract was significantly less in the SLIT vs. the placebo group (P = 0.026). At the end of 6 months, peak expiratory flow (PEF) values in the placebo group was significantly lower than in the SLIT group (P = 0.049). Throughout the treatment period, the SLIT group was found to have less asthma exacerbations than the placebo group (P = 0.007). The provocation concentration causing a 20% drop in forced expired volume in 1 sec did not change throughout the treatment period in either groups. None of the patients reported local or systemic side effects from SLIT. Results of this study suggests that SLIT may be a useful alternative or additional therapy in the treatment of children with asthma/rhinitis due to HDM. PMID- 11416877 TI - Spirometry in children aged 3 to 5 years: reliability of forced expiratory maneuvers. AB - The aim of this study was to evaluate the feasibility and reproducibility of forced expiratory maneuvers during standard spirometric evaluation in preschool children. Among 570 young children attending our laboratory, we retrospectively selected 355 patients (14% 3-4-year-olds, 48% 4-5-year-olds, and 38% 5-6-year olds) who carried out spirometric tests for the first time. The indications for such tests were history of asthma (70%), followed by chronic cough (20%) and other miscellaneous conditions (10%). Eighty-eight, 175, and 92 children performed one, two, and three acceptable tests respectively. Forced expired volume in 1 sec (FEV(1)) and forced vital capacity (FVC) did not differ significantly between attempts in children performing either two or three attempts. Forced expiratory time (FET), i.e., the total time required for the forced expiratory maneuver, was 1.7 +/- 0.1 sec (mean +/- SEM), and was no greater than 1 sec in 21.3% of all tested children. Consequently, FEV(1) does not appear to be well-suited to this age group. Forced expiratory volume in 0.50 and 0.75 sec (FEV(0.5), FEV(0.75)) were thus measured in the group of children performing three attempts (n = 92), and there was no statistical difference between attempts. In 267 children performing two or three tests, the ATS criteria of reproducing FEV(1) and FVC within or = 45 yrs--the cause specific 25-year survival rate was 100%, vs. 62% (at 15 years) (p < 0.0001) in high-risk patients. In stage I and stage II the recurrence-free survival rates at 25 years were 95% and 100% respectively. Risk factors for recurrence were macroscopic (p < 0.0001) but not microscopic local invasion (pT4); stage pN1 (p = 0.0004). Only 1/107 patients initially judged node-negative had a nodal recurrence. FTC (n = 115; mean age 56 yrs; mean follow-up 8 yrs): Cause-related death (n = 8) or serious recurrence (n = 3) occurred in 10/53 grossly invasive FTC, in 1/45 minimally invasive FTC with vascular invasion, and in none of 17 FTC with capsular invasion (CI) alone, under radical treatment (131I) in 75%, 33%, and 12% respectively. 20-year disease-free survival in grossly and in minimally invasive FTC was 78% and 95.5% respectively (p = 0.0007). Patients aged < 45 yrs and patients with minimally invasive FTC with CI alone (all ages) had 100% 20-year disease-free survival vs. 80% (p = 0.013) in the remainder. There was no curable recurrence in FTC. The ratio of grossly invasive FTC decreased (p < 0.0001) during the study period. CONCLUSIONS: Risk-0 groups may be defined and selected for a reduced extent of treatment (PTC pT1-3 N0; FTC < 45 yrs, or CI alone). Older (> or = 45 yrs) patients with PTC in stages I and II have an excellent prognosis (risk 0). With selective (therapeutic) lymphadenectomy the risk of nodal recurrence may be very low in node negative tumours, without use of radioiodine. Meticulous lymphadenectomy is indicated in pN1 tumours with nodal recurrences despite 131I (5/36 patients). The technique of capsular dissection for extracapsular total uni- or bilateral thyroidectomy provides excellent oncological and surgical results. A decrease in the incidence of FTC parallels a decrease in endemic goitre in Switzerland. PMID- 11416890 TI - Non-Hodgkin's lymphoma of the pleural cavity: late complication of artificial pneumothorax for the treatment of pulmonary tuberculosis. PMID- 11416891 TI - Flight, clots and death. PMID- 11416892 TI - CI (critical incidents) vs. CI (corporate identity). PMID- 11416893 TI - Localizing ventricular tachycardia through entrainment. AB - Area(s) of slow conduction are thought to be present within the reentry circuit of most clinically important ventricular tachycardia (VT). To prevent recurrence after ablation of VT late after myocardial infarction, it is desirable to localize and destroy area(s) of slow conduction "critical link" within the reentry circuit. Conventionally, they may be identified by endocardial catheter mapping, continuous electrical activity, mid-diastolic potentials, earliest endocardial activation, pace-mapping etc. However, none of these methods are very specific. Entrainment method may be used to localize the slow conduction zone of reentrant VT. Concealed entrainment is consistent with pacing at a site in the reentry circuit but may also occur at some "bystander" sites that are close to the reentry circuit but are not participating in the circuit itself. During pacing at the slow conduction area of the reentry circuit, the stimulus to QRS (S QRS) interval should equal the electrogram to QRS (EG-QRS) interval during VT. Similarly the post-pacing interval (PPI) approximates the tachycardia cycle length. During pacing at bystander sites, the S-QRS interval may be greater, less than or equal to the EG-QRS interval, depending on the conduction time from the bystander site to the circuit. The PPI, however, always exceed the tachycardia cycle length. In conjunction with concealed entrainment, the use of diastolic potential, double potentials and continuous electrical activity enhances the prediction of radiofrequency termination of post-infarction VT. PMID- 11416894 TI - Benefits of late coronary artery stenting in patients with acute myocardial infarction with and without thrombolytic therapy. AB - BACKGROUND: There are no added benefits when balloon angioplasty is conducted in conjunction with thrombolytic therapy in patients with acute myocardial infarction. The purpose of this study was to determine whether or not thrombolysis has an impact on the outcome of late coronary artery stenting following acute myocardial infarction. METHODS: The outcome of late coronary artery stenting in the infarct-related artery following acute infarction was compared in patients with (68 patients, group A) and without (118 patients, group B) prior thrombolytic therapy. RESULTS: The baseline characteristics were similar in the 2 groups except that total occlusion of the infarct-related artery was more common in group B. The angiographic characteristics of the target lesion were similar in the 2 groups; the procedural success rate was 98% in both groups. There was no subacute thrombosis or other complications in either group. The 6 month follow-up coronary angiography and the restenosis rate was 18% in both groups; the reocclusion rate was 2% in group A and 4% in group B. The increment of the left ventricular ejection fraction was similar in both groups (6% versus 7%). During a follow-up duration of 18 +/- 3 months, the mortality rate was 3% versus 2%, reinfarction 0% versus 1%, recurrent angina 6% versus 4%, and target lesion revascularization by angioplasty 13% versus 13% in group A and B patients, respectively. CONCLUSION: The outcome of late coronary artery stenting following acute myocardial infarction in patients with and without prior thrombolytic therapy was comparable. Significant improvement of left ventricular function was noted in both groups. PMID- 11416895 TI - Neighborhood experience and mental health. AB - BACKGROUND: The purpose of this study was to evaluate individual mental health in the context of their social environment. Specifically, variables across different levels, including a person's attributes, neighborhood experience, and community characteristics, were examined to explore their relationship with that person's mental health. METHODS: Communities in southern Taiwan, including 1 district from Kaohsiung City and 4 townships (hsiang) from Kaohsiung County, were purposively selected to represent 3 different types (urban, suburban, and rural) of communities, after which, by random sampling, 416 subjects were recruited for the interview survey in 1998. A questionnaire containing a measure for mental health (CHQ) and perceptions of neighborhood experience was administered for data collection. At the same time, data derived from governmental archives were collected to indicate the characteristics of a community, and these included population density, community socioeconomic condition, and community stability. RESULTS: Multivariate analysis showed that personal income, community socioeconomic condition, and perceived social support from the neighborhood were statistically significant when associated with a person's mental health status. It seems that influence of an individual's mental health status was affected not only by that person's attributes, but also by the neighborhood within which he/she experiences daily life. CONCLUSION: With respect to the strategy for implementing future mental health promotion programs, a further clarification of the relationship between social conditions and mental health is needed. PMID- 11416896 TI - Effect of carbon dioxide tension in the first three days of life on the development of retinopathy of prematurity. AB - BACKGROUND: This study focuses on the effect of carbon dioxide tension in the first 3 days of life on the development of retinopathy of prematurity (ROP) in mechanically ventilated premature infants. METHODS: From January 1997 to June 1998, 91 premature infants (gestational age 28.8 +/- 2.5 weeks, birth weight 1169 +/- 385 g) born in Chung Gung Children's Hospital requiring mechanical ventilation in the first 72 hours were enrolled in our study. Ophthalmic examination was performed initially at 4-6 weeks of age and then every week if ROP was found and every other week if the result was normal until discharge. These infants were divided into 2 groups: those with and without ROP. Medical records of the highest and lowest blood gas measurements were recorded during the first 3 days of life. To screen for possible risk factors related to the development of ROP within these 2 groups, variables comprising perinatal data were compared. RESULTS: The perinatal factors were compared, whish showed that infants with ROP had a lower mean gestational age (p < 0.001), mean birth body weight (p < 0.001), and a longer duration under mechanical ventilation (p < 0.001). The severity and incidence of ROP were not affected by either hypercapnia or hypocapnia; even with the superimposed effect of hyperoxia, the effect was minimal. CONCLUSION: Hypercarbia or hypocarbia in the first 3 days of mechanically ventilated preterm neonates does not affect the subsequent development of retinopathy of prematurity. However, a lower gestational age and birth weight as well as longer-duration mechanical ventilation might be independent predictors for the occurrence of ROP. The role of PaCO2 in the pathophysiology of retinopathy of prematurity remains undetermined, and further investigation in the future is warranted. PMID- 11416897 TI - Pancreatic pseudocysts in children. AB - BACKGROUND: Pancreatic pseudocysts in children are uncommon. The purpose of this study was to investigate the clinical course, image findings (with emphasis on sonograms), and outcome of 12 patients with pancreatic pseudocysts. METHODS: From January 1986 to May 2000, 12 patients with a diagnosis of pancreatic pseudocysts were encountered in our hospital. There were 8 males and 4 females with ages ranging from 3 to 18 years. RESULTS: The etiology was blunt abdominal trauma in all 12 cases. The cysts were detected from 3 to 57 days after injury, and the sizes ranged from 0.6 to 16 cm. Six patients received surgical treatment. External drainage was performed in 3 cases, subtotal pancreatectomy in 1, cystogastrostomy in 2. Initial serum amylase level correlated with neither the time to normalization (r = 0.354, p = 0.268), cystic appearance after trauma (r = 0.029, p = 0.933), nor resolution (r = 0.322, p = 0.309). Sonographic follow-up revealed that the disappearance of cysts was noted from 11 days to 10 months following injury. The size of the pseudocysts correlated with neither the time of detection (r = 0.284, p = 0.371) nor disappearance (r = -0.175, p = 0.586). CONCLUSION: Serial sonogram examinations play an important role in monitoring the progress of pancreatic pseudocysts, which may develop even 57 days after injury. The size of pseudocysts correlated with neither the times of detection nor disappearance, and the initial serum amylase level correlated with neither the time to normalization nor cystic appearance after trauma. PMID- 11416898 TI - Perfluorodecalin condensation on posterior surface of lens capsule and corneal endothelium during vitrectomy. AB - BACKGROUND: High-density vitreous substitutes, including perfluorodecalin, are widely used in current vitreoretinal surgery. We report 4 cases of perfluorodecalin condensation on the posterior capsule of the lens or corneal endothelium during vitrectomy. To the best of our knowledge, no such phenomenon has ever been reported. METHODS: This study consisted of 4 patients with retinal detachment and large retinal tears. All patients were men. All of the patients underwent pars plana vitrectomy, perfluorodecalin injection, endolaser retinopexy and air-fluid exchange. Two patients received additional surgery for scleral buckling. RESULTS: Perfluorodecalin condensation was observed on the posterior surface of the lens capsule in one phakic patient and on the endothelium in three aphakic patients during vitrectomy. The condensation obscured the image of the fundus and made air-fluid exchange difficult. The condensation could be dissipated by first removing the perfluorodecalin as completely as possible, and then by lowering the intraocular pressure below the vapor pressure of perfluorodecalin. CONCLUSIONS: Perfluorodecalin condensation may occur when perfluorodecalin is not properly used. To avoid forming condensation, high density vitreous substitutes should never be injected into eyes that are completely air-filled, prolonged air-fluid exchanges should be avoided, and the intraocular pressure should always be kept at a higher level during air-fluid exchange. PMID- 11416899 TI - Unilateral congenital third cranial nerve palsy with central nervous system anomalies: report of two cases. AB - The third nerve palsies are not uncommon in adults and are usually the result of vascular or compressive lesions. However, they are uncommon in children and are usually congenital. Congenital oculomotor nerve palsies were generally thought to exist in isolation without associated abnormalities; however, there have been conflicting reports in the literature as to the associations of congenital oculomotor palsies. In this report, we present 2 cases of congenital third cranial nerve palsy with neurological deficits and central nervous system abnormalities. The first patient had initial presentations of inequality in pupil size, exodeviation of the affected eye, and extreme hypotropia of the contralateral eye, which was also complicated by profound neurological deficits. Magnetic resonance imaging (MRI) scans disclosed hypoplasia of the right midbrain and corpus callosum. The second patient presented with an exodeviated right eye, and pupils non-reactive to light since birth. Computed tomography scans showed ventricular dilatation and absence of the septum pellucidum, which is compatible with septo-optic dysplasia (de Morsier syndrome). The clinical courses are presented, and we also review the literature concerning congenital third cranial nerve palsy associated with central nervous system anomalies. PMID- 11416900 TI - Thoracoscopic retrieval of metal rods after penetrating chest injury: case report. AB - Thoracoscopy has been used for both diagnostic and therapeutic purposes. Its use in the trauma field is rapidly increasing. Here, we present a case of chest trauma that was successfully treated using the thoracoscopic approach. A 43-year old male patient was brought to our emergency room with a severe right chest wall penetrating metal-rod injury, which had occurred after falling from a height. The chest X-ray showed an upper right lung lobe injury. Video-assisted thoracoscopic surgery (VATS) was performed for diagnosis of any other associated injury and for management of the penetrating injury. The prognosis was good. We believe that minimal thoracoscopic surgery is an alternative way that provides a more rapid diagnosis, and a less-invasive and safe operation for acute chest trauma patients. PMID- 11416901 TI - A second deciduous molar impacted in right maxillary sinus: a long-term follow up. AB - Primary tooth impaction is quite rare during the development of primary dentition. Various factors contribute to the impaction of a deciduous tooth, including malposition and malformation of the tooth bud, dentomaxillofacial disharmony, tumors, and cysts. This case was a 6-year-old female with complete left cleft lip/palate. A maxillary right second deciduous molar impacting at a high level approximate to the maxillary sinus was noted on the initial visit. To prevent surgical trauma to the surrounding tissue and succedaneous teeth, regular follow-up with radiographic examination was taken every 2 years. After a 7-year long-term follow-up, the impacted second primary molar in the right maxillary sinus had almost completely been resorbed with only about 10% of the original structure left. To the present, the resorption of the impacted deciduous molar seemed to have continued with no major pathological changes. Periodic radiographic examination is strongly advised until the total resorption process is completed. PMID- 11416902 TI - Symptomless cyst formation at the location of a biloma resolved with a single aspiration: case report. AB - A biloma is an encapsulated bile collection outside the biliary tree. The underlying causes include iatrogenic, traumatic, and spontaneous injury of the biliary tree. Its diagnosis is based upon clinical history, imaging studies, and needle aspiration with chemical analysis of the fluid. Biloma usually presents as a simple or septated cyst or a cyst with internal contents. The symptomatic biloma if left untreated may result in significant morbidity and even mortality. In past reports, the mainstay of management relied on bile flow diversion and/or bile drainage to achieve elimination of bile as well as complete resolution of the cyst. We report on a 58-year-old woman with a 9.0 x 5.4 cm cyst between the right liver and the upper role of the kidney after wedge resection for a hepatoma in the right lobe. She suffered from abdominal pain and fever. A single percutaneous needle aspiration with chemical analysis confirmed the diagnosis of biloma and relieved her symptoms. The biloma vanished immediately. However, a recurrent symptomless cyst was detected at the same site 6 months later. Its content was proven to be clear transudate, and it was left untreated. If a biloma has no communication with the biliary tree, and the proximal biliary tree has neither stricture nor stones, percutaneous aspiration alone may achieve resolution. PMID- 11416903 TI - Bibliography Current World Literature. Neuroscience. PMID- 11416904 TI - Bibliography Current World Literature. Gastrointestinal system. PMID- 11416905 TI - The signature of scrapie: differences in the PrP genotype profile of scrapie affected and scrapie-free UK sheep flocks. AB - The amino-acid sequence of the PrP protein plays an important role in determining whether sheep are susceptible to scrapie. Although the genetics of scrapie susceptibility are now well understood, there have been few studies of the PrP gene at the population level, especially in commercially farmed sheep. Here we describe the PrP genetic profiles of the breeding stock of four UK sheep flocks, comprising nearly 650 animals in total. Two flocks had been scrapie affected for about eight years and two were scrapie free. Scrapie-resistant PrP genotypes predominated in all flocks but highly susceptible genotypes were present in each case. The distribution of PrP genotypes was similar in the scrapie-affected and scrapie-free flocks. The former, however, showed a slight but significant skew towards more susceptible genotypes despite their previous losses of susceptible sheep. Surprisingly, this skew was apparent in younger, but not older, sheep. We suggest that these patterns may occur if sheep flocks destined to become scrapie affected are predisposed by a genetic profile skewed towards susceptibility. The age structure of the scrapie-affected flocks suggests that the number of losses attributable directly or indirectly to scrapie considerably exceeds that recognized by the farmers, and also that significant losses may occur even in sheep of a moderately susceptible genotype. Similar patterns were not detected in the scrapie-free flocks, indicating that these losses are associated with scrapie infection as well as genotype. PMID- 11416906 TI - The cause of parasitic infection in natural populations of Daphnia (Crustacea: Cladocera): the role of host genetics. AB - Disease patterns in nature may be determined by genetic variation for resistance or by factors, genetic or environmental, which influence the host-parasite encounter rate. Elucidating the cause of natural infection patterns has been a major pursuit of parasitologists, but it also matters for evolutionary biologists because host resistance genes must influence the expression of disease if parasite-mediated selection is to occur. We used a model system in order to disentangle the strict genetic component from other causes of infection in the wild. Using the crustacean Daphnia magna and its sterilizing bacterial parasite Pasteuria ramosa, we tested whether genetic variation for resistance, as determined under controlled conditions, accounted for the distribution of infections within natural populations. Specifically, we compared whether the clonally produced great-granddaughters of those individuals that were infected in field samples (but were subsequently 'cured' with antibiotics) were more susceptible than were the great-granddaughters of those individuals that were healthy in field samples. High doses of parasite spores led to increased infection in all four study populations, indicating the importance of encounter rate. Host genetics appeared to be irrelevant to natural infection patterns in one population. However, in three other populations hosts that were healthy in the field had greater genetic-based resistance than hosts that were infected in the field, unambiguously showing the effect of host genetic factors on the expression of disease in the wild. PMID- 11416907 TI - Fitting host-parasitoid models with CV2 > 1 using hierarchical generalized linear models. AB - The powerful general Pacala-Hassell host-parasitoid model for a patchy environment, which allows host density-dependent heterogeneity (HDD) to be distinguished from between-patch, host density-independent heterogeneity (HDI), is reformulated within the class of the generalized linear model (GLM) family. This improves accessibility through the provision of general software within well known statistical systems, and allows a rich variety of models to be formulated. Covariates such as age class, host density and abiotic factors may be included easily. For the case where there is no HDI, the formulation is a simple GLM. When there is HDI in addition to HDD, the formulation is a hierarchical generalized linear model. Two forms of HDI model are considered, both with between-patch variability: one has binomial variation within patches and one has extra binomial, overdispersed variation within patches. Examples are given demonstrating parameter estimation with standard errors, and hypothesis testing. For one example given, the extra-binomial component of the HDI heterogeneity in parasitism is itself shown to be strongly density dependent. PMID- 11416908 TI - Dynamics of a feline virus with two transmission modes within exponentially growing host populations. AB - Feline panleucopenia virus (FPLV) was introduced in 1977 on Marion Island (in the southern Indian Ocean) with the aim of eradicating the cat population and provoked a huge decrease in the host population within six years. The virus can be transmitted either directly through contacts between infected and healthy cats or indirectly between a healthy cat and the contaminated environment: a specific feature of the virus is its high rate of survival outside the host. In this paper, a model was designed in order to take these two modes of transmission into account. The results showed that a mass-action incidence assumption was more appropriate than a proportionate mixing one in describing the dynamics of direct transmission. Under certain conditions the virus was able to control the host population at a low density. The indirect transmission acted as a reservoir supplying the host population with a low but sufficient density of infected individuals which allowed the virus to persist. The dynamics of the infection were more affected by the demographic parameters of the healthy hosts than by the epidemiological ones. Thus, demographic parameters should be precisely measured in field studies in order to obtain accurate predictions. The predicted results of our model were in good agreement with observations. PMID- 11416909 TI - The control of morph development in the parasitic nematode Strongyloides ratti. AB - The parasitic nematode Strongyloides ratti has a complex life cycle. The progeny of the parasitic females can develop into three distinct morphs, namely directly developing infective third-stage larvae (iL3s), free-living adult males and free living adult females. We have analysed of the effect of host immune status (an intra-host factor), environmental temperature (an extra-host factor) and their interaction on the proportion of larvae that develop into these three morphs. The results are consistent with the developmental decision of larvae being controlled by at least two discrete developmental switches. One is a sex-determination event that is affected by host immune status and the other is a switch between alternative female morphs that is affected by both host immune status and environmental temperature. These findings clarify the basis of the life cycle of S. ratti and demonstrate how such complex life cycles can result from a combination of simple developmental switches. PMID- 11416910 TI - Optimization of adult performance determines host choice in a grass miner. AB - Models and empirical studies on host selection in plant-insect, algae-amphipod, host-parasite and prey-predator systems assume that oviposition preference is determined by the quality of the oviposition site for offspring development. According to the oviposition-preference-offspring-performance hypothesis, oviposition-preference hierarchy should correspond to host suitability for offspring development because females maximize their fitness by optimizing offspring performance. We show, we believe for the first time, that adult feeding site and related adult performance may explain most of the variation in adult feeding and oviposition site selection of an oligophagous grass miner, Chromatomyia nigra (Diptera). This study advances our understanding of the complex interactions between plants and herbivores because it shows that host preference patterns are not only shaped by the optimization of offspring performance, as previously assumed, but also by the optimization of adult performance. PMID- 11416911 TI - Intrasexual mounting in the beetle Diaprepes abbreviatus (L.). AB - The weevil Diaprepes abbreviatus shows three kinds of same-sex mountings: males mount other unpaired males, males mount males already engaged in copulation and females mount other females. Four hypotheses were evaluated in order to explain same-sex matings by males: (i) female mimicry by inferior males, (ii) dominance of larger males which affects the behaviour of small males, (iii) sperm transfer in which smaller males gain some reproductive success by 'hitchhiking' their sperm with the sperm of larger males, and (iv) poor sex recognition. Data from mate choice and sperm competition experiments rejected the female mimicry, dominance and sperm transfer hypotheses and supported the poor sex recognition hypothesis. We tested three hypotheses in order to explain female mounting behaviour: (i) females mimic male behaviour in order to reduce sexual harassment by males, (ii) females mount other females in order to appear larger and thereby attract more and larger males for mating, and (iii) female mimicry of males. The results of our mate choice experiments suggested that the female mimicry of males hypothesis best explains the observed female mounting behaviour. This result is also consistent with the poor sex recognition hypothesis which is the most likely explanation for male and female intrasexual mating behaviour in many insect species. PMID- 11416912 TI - Anticipatory parental care: acquiring resources for offspring prior to conception. AB - Many organisms acquire and defend resources outside the breeding season and this is thought to be for immediate survival and reproductive benefits. Female red squirrels (Tamiasciurus hudsonicus) acquire traditional food cache sites up to four months prior to the presence of any physiological or behavioural cues associated with mating or offspring dependency. They subsequently relinquish these resources to one of their offspring at independence (ten months later). We experimentally show that acquisition of these cache sites cannot be explained by conventional arguments such as enhanced survival of the parent or increased reproductive output. Instead this behaviour functions as a form of parental care. PMID- 11416913 TI - Trade-off between mating opportunities and parental care: brood desertion by female Kentish plovers. AB - Why do some parents care for their young whereas others divorce from their mate and abandon their offspring? This decision is governed by the trade-off between the value of the current breeding event and future breeding prospects. In the precocial Kentish plover Charadrius alexandrinus females frequently, but not always, abandon their broods to be cared for by their mate, and seek new breeding partners within the same season. We have shown previously that females' remating opportunities decline with date in the season, so brood desertion should be particularly favourable for early breeding females. However, the benefits are tempered by the fact that single-parent families have lower survival expectancies than those where the female remains to help the male care for the young. We therefore tested the prediction that increasing the value of the current brood (by brood-size manipulation) should increase the duration of female care early in the season, but that in late breeders, with reduced remating opportunities, desertion and thus the duration of female care should be independent of current brood size. These predictions were fulfilled, indicating that seasonally modulated trade-offs between current brood value and remating opportunities can be important in the desertion decisions of species with flexible patterns of parental care. PMID- 11416914 TI - Rate of moult affects feather quality: a mechanism linking current reproductive effort to future survival. AB - Life-history theory proposes that costs must be associated with reproduction. Many direct costs are incurred during breeding. There is also evidence for indirect costs, incurred after breeding, which decrease survival and future reproductive success. One possible indirect cost identified in birds is that breeding activity in some way compromises plumage quality in the subsequent moult. Here we propose a mechanism by which this could occur. Breeding activity delays the start of moult. Birds that start to moult later also moult more rapidly--an effect of decreasing daylength. Could this result in poorer quality plumage? We kept two groups of male European starlings, Sturnus vulgaris, one on constant long days and the other on decreasing daylengths from the start of moult. Decreasing daylengths reduced the duration of moult from 103 +/- 4 days to 73 +/- 3 days (p < 0.0001). Newly grown primary feathers of birds that moulted fast were slightly shorter, weighed less (p < 0.05) and were more asymmetrical. They had a thinner rachis (p < 0.005), were less hard (p < 0.01) and less rigid (p < 0.05). They were also less resistant to wear so that differences in mass and asymmetry increased with time. There was no difference in Young's modulus. Poorer quality plumage will lead to decreased survival due to decreased flight performance and increased thermoregulatory costs. Thus, reproduction incurs costs through a mechanism that operates after the end of breeding. PMID- 11416915 TI - Variation in the volume of zebra finch song control nuclei is heritable: developmental and evolutionary implications. AB - In many songbird species, females prefer males that sing a larger repertoire of syllables. Males with more elaborate songs have a larger high vocal centre (HVC) nucleus, the highest structure in the song production pathway. HVC size is thus a potential target of sexual selection. Here we provide evidence that the size of the HVC and other song production nuclei are heritable across individual males within a species. In contrast, we find that heritabilities of other nuclei in a song-learning pathway are lower, suggesting that variation in the sizes of these structures is more closely tied to developmental and environmental differences between individuals. We find that evolvability, a statistical measure that predicts response to selection, is higher for the HVC and its target for song production, the robustus archistriatalis (RA), than for all other brain volumes measured. This suggests that selection based on the functions of these two structures would result in rapid major shifts in their anatomy. We also show that the size of each song control nucleus is significantly correlated with the song related nuclei to which it is monosynaptically connected. Finally, we find that the volume of the telencephalon is larger in males than in females. These findings begin to join theoretical analyses of the role of female choice in the evolution of bird song to neurobiological mechanisms by which the evolutionary changes in behaviour are expressed. PMID- 11416916 TI - Maternal and grandmaternal age influence offspring fitness in Drosophila. AB - The influence of maternal and grandmaternal age on progeny egg-to-adult viability was assessed in Drosophila serrata. Viability in progeny decreased with increasing maternal age. The potential for cumulative age effects was investigated in two environments, one of which involved nutrient and cold stress. Environment influenced viability and female age influenced progeny egg-to-adult viability across one generation. The influence on viability was cumulative across two generations. Females from old mothers, who also had old grandmothers, had the lowest viability in both environments. Grandmaternal effects were associated with a decrease in egg hatch rate whereas maternal effects also involved larval-to adult viability. The age of the mother and grandmother should be taken into account when evaluating life-history traits in Drosophila. PMID- 11416917 TI - Sexual dimorphism matches photoreceptor performance to behavioural requirements. AB - Differences in behaviour exist between the sexes of most animal species and are associated with many sex-specific specializations. The visual system of the male housefly is known to be specialized for pursuit behaviour that culminates in mating. Males chase females using a high-acuity region of the fronto-dorsal retina (the 'love spot') that drives sex-specific neural circuitry. We show that love spot photoreceptors of the housefly combine better spatial resolution with a faster electrical response, thereby allowing them to code higher velocities and smaller targets than female photoreceptors. Love spot photoreceptors of males are more than 60% faster than their female counterparts and are among the fastest recorded for any animal. The superior response dynamics of male photoreceptors is achieved by a speeding up of the biochemical processes involved in phototransduction and by a tuned voltage-activated conductance that boosts the membrane frequency response. These results demonstrate that the inherent plasticity of phototransduction facilitates the tuning of the dynamics of visual processing to the requirements of visual ecology. PMID- 11416918 TI - Olfactory coding in a simple system: adaptation in Drosophila larvae. AB - Drosophila melanogaster larvae were pre-stimulated with high concentrations of six homologous alcohols (C4-C9) and then tested for adaptation and cross adaptation using these same alcohols, four aliphatic n-acetates and three acids. Pre-stimulation with hexanol effectively reduced to zero (abolished) test responses to all six alcohols, whereas test responses to hexanol were only affected by pre-stimulation with hexanol. This substance appears to play a fundamental role in the organization of the larval olfactory system. Test responses to butanol and pentanol, and the effect of pre-stimulation with butanol and pentanol, were not significantly different, indicating that they are sensory equivalents. Heptanol, octanol and nonanol induce a complex set of responses among one another. Cross-adaptation between functional groups was observed, in particular following pre-stimulation with hexanol, but there was also evidence that functional groups are coded separately. A model of olfactory processing in the fruitfly maggot is presented that explains the data and provides predictions for future anatomical, genetic and electrophysiological studies. PMID- 11416919 TI - The quantitative genetic basis of offspring solicitation and parental response in a passerine bird with biparental care. AB - The coevolution of parental investment and offspring solicitation is driven by partly different evolutionary interests of genes expressed in parents and their offspring. In species with biparental care, the outcome of this conflict may be influenced by the sexual conflict over parental investment. Models for the resolution of such family conflicts have made so far untested assumptions about genetic variation and covariation in the parental resource provisioning response and the level of offspring solicitation. Using a combination of cross-fostering and begging playback experiments, we show that, in the great tit (Parus major), (i) the begging call intensity of nestlings depends on their common origin, suggesting genetic variation for this begging display, (ii) only mothers respond to begging calls by increased food provisioning, and (iii) the size of the parental response is positively related to the begging call intensity of nestlings in the maternal but not paternal line. This study indicates that genetic covariation, its differential expression in the maternal and paternal lines and/or early environmental and parental effects need to be taken into account when predicting the phenotypic outcome of the conflict over investment between genes expressed in each parent and the offspring. PMID- 11416920 TI - Implementing the revised American Psychiatric Association practice guideline for the treatment of patients with eating disorder. AB - Practice guidelines aspire to be authoritative statements regarding the state of the art in quality care for various clinical problems. The American Medical Association Partnership has set forth stringent parameters for guideline development by professional organizations, and these have been followed by the American Psychiatric Association in creating its practice guidelines for the treatment of patients with eating disorders. The revised edition, published in January 2000, benefitted from extensive input from a wide array of psychiatrists, psychologists, pediatricians, and other recognized experts who blended together available evidence-based practice with a considerable amount of clinical experience and consensus. These guidelines are useful for practitioners, students, and health-resource managers. Future research will continue to lead to constant upgrades. PMID- 11416921 TI - Epidemiology and mortality of eating disorders. AB - Although considerable progress has been made over the past 2 decades in the field of eating-disorder epidemiology, there is room for improvement. Eating disorder diagnoses for children, need assessment, prospective longitudinal studies, studies of natural cause, treatment dose-effect studies, and bias are all areas that need careful consideration. Clinicians should consider establishing a Cochrane database for eating disorders. The mortality studies reviewed in this article are dominated by small, sparse, unbalanced data sets, and publication bias and bias caused by loss to follow-up limits the validity of the reported findings. Some methodologic advice is given in the hope that some of the more obvious pitfalls can be avoided in future research. PMID- 11416922 TI - The evolving genetic foundations of eating disorders. AB - Data described earlier are clear in establishing a role for genes in the development of eating abnormalities. Estimates from the most rigorous studies suggest that more than 50% of the variance in eating disorders and disordered eating behaviors can be accounted for by genetic effects. These high estimates indicate a need for studies identifying the specific genes contributing to this large proportion of variance. Twin and family studies suggest that several heritable characteristics that are commonly comorbid with AN and BN may share genetic transmission with these disorders, including anxiety disorders or traits, body weight, and possibly major depression. Moreover, some developmental research suggests that the genes involved in ovarian hormones or the genes that these steroids affect also may be genetically linked to eating abnormalities. Molecular genetic research of these disorders is in its infant stages. However, promising areas for future research have already been identified (e.g., 5-HT2A receptor gene, UCP-2/UCP-3 gene, and estrogen receptor beta gene), and several large-scale linkage and association studies are underway. These studies likely will provide invaluable information regarding the appropriate phenotypes to be included in genetic studies and the genes with the most influence on the development of these disorders. PMID- 11416923 TI - Clinical implications of brain imaging in eating disorders. AB - Imaging studies have greatly improved the understanding of the pathology and physiology of psychiatric disorders, such as schizophrenia, affective disorders, obsessive-compulsive disorder, and Tourette syndrome. In the past few years, several neuroimaging studies have concentrated on patients with eating disorders. Although the number of studies is small compared with studies of other psychiatric disorders, the results are beginning to highlight potential areas in the brain that may lead to a better understanding of these disorders. Much research still is needed, and replication of results across centers is needed. The brain is an extremely complex organ; that eating disorders are a result of abnormalities in one specific area of the brain is unlikely. More likely is that several components of the brain have a role, including cortex and subcortical regions and involvement of several neurochemical pathways and circuits within the brain. Further studies are needed in this exciting development of research about eating disorders. PMID- 11416924 TI - Dysfunctional eating in the eating disorders. AB - Aberrant eating patterns in the eating disorders have been observed across various laboratory-based and clinical studies. It is now clear that problems in experiencing and expressing hunger, appetite, and satiety in anorexia and bulimia nervosa are likely to perpetuate the disorders once established. Whether problems in appetite regulation are primary or secondary to the development of the disorders is unknown. In studies examining indices of appetite regulation after treatment, there still remain significant levels of eating abnormality. This suggests that the main goals of treatment, including restoration of body weight in anorexia nervosa, abstaining from dieting in anorexia or bulimia nervosa, and reducing or abstaining from binge eating, do not correct some features of abnormal eating. The efficacy of nutritional counseling and specific nutritional management programs have been tested, and these seem to produce positive outcomes in improving eating behavior. Direct behavioral interventions to change eating patterns also have been examined, and these too seem to produce benefits that may be incorporated into CBT. Greater collaboration and cooperation between researchers and clinicians in addressing dysfunctional eating in the eating disorders will highlight improvements in treatment for identifiable eating abnormalities and will further the understanding of the human appetite system. PMID- 11416925 TI - The role of personality in the onset of eating disorders and treatment implications. AB - The relationship of personality and eating disorders is largely unclear. The development of well-specified conceptual models of this relationship, developments in family history, behavior genetic and prospective longitudinal research methodology, and careful consideration of how trait constructs may interact with state variables to produce or maintain eating disorders all will help to advance this area of research. PMID- 11416926 TI - Eating disorders in children. Diagnosis and age-specific treatment. AB - The symptoms of children with eating disorders change as these children age. In each age group, the diagnosis, evaluation, and treatment should be tailored to the developmental level of the child and should address the needs of the child's family. With early diagnosis and treatment comes the opportunity to avoid long term sequelae of chronic eating disorders in adults. Pediatric eating-disordered patients are not "miniature adults" who can be treated by an adult protocol or unit that does not address their developmentally specific needs. With appropriate evaluation and treatment, eating disorders can be stopped in childhood. PMID- 11416927 TI - A slimming program for eating disorders not otherwise specified. Reconceptualizing a confusing, residual diagnostic category. AB - This study suggests that the category of EDNOS as currently defined is overly broad, representing many cases that could be more helpfully subsumed within AN or BN diagnostic criteria without changing the essential features of these categories but by rethinking the currently overly restrictive, perhaps research derived criteria. The reconceptualizing of AN as a syndrome resulting from a decrement between setpoint versus illness-driven final weight avoids the inherent problems of imposing a category on a dimension. A rethinking of AN suggests that a specific female-only abnormality of reproductive hormone functioning, 3 months of amenorrhea, is too restrictive. Instead, a more encompassing criterion recognizing the multiple medical, social, and psychologic functional impairments that result from substantial starvation would be appropriate in its place. Clinicians who otherwise confidently treat AN and BN patients would welcome the clearer diagnostic categorization of the potentially confusing EDNOS category. Third party payers who currently, albeit wrongly, exclude EDNOS diagnoses from insurance payment, would have less difficulty with a smaller group of EDNOS. In summary, the currently overly broad category of EDNOS as currently used would benefit from a thoughtful dieting regimen. PMID- 11416928 TI - Matching patient variables to treatment intensity. The continuum of care. AB - This article has reviewed what is currently known regarding the relationship between specific patient variables and treatment response in AN, BN, and BED. Matching patient variables to treatment intensity remains an important and fruitful area for future research. There is a need for established guidelines for clinicians regarding the choice of the appropriateness of treatment settings and type of interventions delivered in those settings. These guidelines should be evidence based, with clear clinical indicators for each of the recognized eating disorders and their subclinical variants. PMID- 11416929 TI - Basic principles for applying cognitive-behavioral therapy to anorexia nervosa. AB - Cognitive-behavioral therapy continues to be one of the recommended evidenced based interventions in the treatment of eating disorders. Cognitive-behavioral therapy is clearly of benefit in anorexia and bulimia nervosa, can be learned in a reasonable time, and is guided by manuals on cognitive-behavioral therapy. This article looks at theoretic and clinical interventions in the treatment of anorexia nervosa and suggests strategies related to effective psychotherapeutic treatment that can be used across a continuum of care. PMID- 11416930 TI - The crucial role of psychodynamic understanding in the treatment of eating disorders. AB - Psychodynamic interventions for eating-disordered patients can provide a unique pathway to understanding the illness. Contemporary practice strives to integrate insights derived from drive theory, ego psychology, object relations, self psychology, relational and attachment theories, and intersubjectivity to claim a more thorough understanding of the antecedents of the eating-disorder symptoms. Placing emphasis on the unique personal history of the individual and finding a safe haven in which to process that history are cornerstones of psychodynamic treatment embraced by each of these schools of thought. This article has emphasized the importance of creating a safe space for these patients to speak, processing the most salient affects that arise in the therapeutic dyad, assessing potential transference and counter-transference paradigms, and assisting these patients in mastering symptomatic behaviors by making healthier identifications with their therapists. Although the selected topics merely sketch how psychodynamic treatment may apply in some cases, it is hoped that they will whet the appetite for a more sustained inquiry for readers. Contemporary psychoanalysis aims to be more "user-friendly" for patients and treaters and eschews a "one interpretation fits all" approach to any person suffering from a particular diagnosis. Attachment theory, infant and developmental studies, and in depth outcomes research are shaping the way that psychoanalytically informed treatments are carried out. Within the next decade, insights derived from these kinds of psychoanalytic research will be even more concretely and meaningfully used in the treatment of eating disorders and other Axis I conditions. PMID- 11416931 TI - Combining pharmacotherapy and psychotherapy in the treatment of patients with eating disorders. AB - The available treatment literature supports a role for medication management in the treatment of both AN and BN. The data on BN are clearer and suggest that antidepressant therapy--fluoxetine being the drug most widely studied--is superior to treatment with placebo but less effective than CBT alone, with one such study suggesting that the combination may provide optimal treatment. Specific recommendations as to when to add or not add antidepressants to CBT have been made, although the rules suggested here have yet to be empirically tested. Although the data on AN are much more limited, information available suggests a lack of efficacy for SSRIs in patients with AN at low weight and considerable use for SSRIs when used in combination with psychotherapy for patients with AN following weight recovery. Where do we go from here? Several pressing issues require careful study. First, in the case of patients with AN, can other agents, in particular the new atypical antipsychotics, be useful in treating patients when they are at low weight? In terms of relapse prevention, can the available findings indicating a role for antidepressants in relapse prevention be replicated, and, if so, can predictor variables that are associated with antidepressant response be identified? In the case of BN, clinicians need to know more about the best possible way to sequence interventions. It has been proposed to add medication to CBT early in treatment if the response to CBT alone is thought to be inadequate. However, other models should be considered, such as stepped-care models in which self-help manuals are used in conjunction with medications. The advantage of these interventions is they could be made more widely available than CBT, which requires a specialist's care. Also, several other new agents, such as sibutramine, which is a drug with serotonin and norepinephrine reuptake inhibition effects, should be tested empirically in subjects with BN, given their pharmacologic profiles. PMID- 11416932 TI - Binge-eating disorder and obesity. A combined treatment approach. AB - Patients presenting with both obesity and BED face multiple challenges: normalizing their eating, improving their physical health, and working to enhance their own acceptance of their body image. In an effort to feel better about themselves, they often have become trapped in a cycle of desperately attempting to diet, then losing control, binge eating, and gaining even more weight. Several psychological and pharmacologic treatment approaches have been used in this population. Most suppress binge eating in the short term, and some seem promising in the long term as well. However, sustained weight loss remains a largely unrealized goal. More recently, BED treatment programs have attempted to address these goals sequentially or in combination. In either approach, it is clear that adopting a long-term focus and promoting enhanced self-acceptance, which have so often been missing from these patients' previous attempts at recovery, are important tasks of treatment that are likely to lead to beneficial lifestyle changes and long-term improvements in physical and psychological health. PMID- 11416933 TI - Involuntary treatment in anorexia nervosa. AB - It is difficult to predict, on first contact with patients with AN, whether a compulsory admission to hospital may become necessary to protect their lives and health. There are only tentative pointers so far to an entrenched avoidance of treatment: (1) components of a disordered personality associated with a history of childhood physical or sexual abuse or previous episodes of self-harm and (2) the presence of a more severe illness, suggested by numerous previous admissions. An involuntary admission is likely to be beneficial at least in the short term, as shown by a gratifying weight gain, although a longer period of inpatient stay may be necessary. Patients who have required compulsory detention are at a considerable risk in the long-term as shown by their high mortality rates. It is, therefore, essential to organize long-term observation for all patients who required involuntary admission for AN. A compulsory admission for AN does not require compulsory treatment, such as forced feeding by NGT or other intrusive methods. Clinicians who contemplate a compulsory admission for a seriously ill anorexic patient might therefore question the advantages provided by the detention. First, clinicians can be assured that it should be possible to induce a satisfactory weight gain through persistent nursing methods without running the risk of these patients discharging themselves. With inpatients, the goals are nearly always attained, although the admission may be longer than average. Not only do these patients' nutrition improve vastly, but also they are likely to show improvements in their mental state. Secondly, these patients are likely to learn that the professional staff, their families, and outside agencies take their illness very seriously, even if patients themselves do not seem to. This is particularly evident when patients appeal to a Mental Health Review Tribunal for release. They attend the proceedings and hear the evidence presented by their psychiatrists, the nursing staff, and their nearest relatives. The tribunal usually sustains the compulsory admission: the patient may be initially distressed, but in the long run the experience is generally therapeutic. Finally, compulsory admission permits more stringent forms of supervision. For example, patients who vomit may have legitimately restricted access to bathrooms. Patients addicted to exercise may be rationed to sensibly short periods of walking daily. Patients who are extremely anxious or overactive may be required to take appropriate tranquilizing or sedating drugs, such as one of the benzodiazepine drugs. Clinicians sometimes are reluctant to resort to compulsory admission because of a fear of damaging the therapeutic relationship with their patients. Clinical observations, however, point to the converse being the case, as shown in several studies. Tiller et al maintain: "Compulsory treatment may be an act of compassion: it shows that professionals recognize the severity of the illness and that they are prepared to contain the anxieties provoked by weight gain. Often the patients and their families are immensely relieved to hand over the responsibility, temporarily, to the professional team." PMID- 11416934 TI - Characteristics of an ideal psychotherapist for eating-disordered patients. AB - Psychotherapy is an essential component of the treatment of eating disorders. A fully competent ideal psychotherapist needs a body of knowledge; personality characteristics of nonpossessive warmth and perseverance; training in scientific, manual-based psychotherapy; and experience, including supervision. The formal discipline of the therapist is less important than are these qualifications. Ideal means fully competent, not perfect. An ideal therapist is a crucial component in the treatment of eating-disordered patients. PMID- 11416935 TI - Osteoporosis in young people. Research and treatment in eating disorders. AB - We still have much to learn about BMD problems in eating disorders. Much progress has been made in the past 10 years; most clinicians and many patients and their families are now aware of the problem. More research is crucial, however, the authors suggest focusing on three areas: 1. Treatment and prevention: Such studies are difficult to conduct for similar reasons to the difficulties in conducting treatment trials of therapy for AN. First, the relative rarity of the condition makes it difficult to recruit subjects; second, drop-out rates are higher because of ambivalence; and third, the population is heterogeneous both in terms of symptoms and cause. 2. Better understanding of bone turnover in AN. More studies are needed to examine turnover of bone in patients with AN using biochemical markers. In particular, prospective studies are needed to examine the effects of refeeding, weight gain, and treatments such as calcium supplementation. 3. Long-term course of bone density. It would be particularly instructive to examine this in individuals with a short or long history of AN. It would also be useful to study women approaching menopause who had an episode of AN in their teens or early twenties compared with women who were of normal weight during this period. PMID- 11416936 TI - The consequences and costs of the eating disorders. AB - Eating disorders are severe, relatively chronic conditions that are associated with comorbid psychopathology and adverse medical conditions. The death rate for patients with AN is the highest among psychiatric conditions, with high suicide rates and deaths from physiologic causes. In addition, the costs of therapy for AN are higher than those for schizophrenia. Although somewhat less chronic, BN and binge-eating disorder are costly conditions to treat, similar to or more expensive than the costs for the treatment of OCD. Although antidepressant medication seems to be the most cost-effective treatment in the short term, given the higher relapse rates with antidepressants, it seems that, in the end, CBT may be the most cost-effective approach to the treatment of BN. It is possible that similar figures would occur for binge-eating disorder. The issue of the comparative cost-effectiveness of various treatments for psychiatric disorders has been neglected in the research literature to date. It is important that large scale RCTs add a sophisticated cost-effectiveness analysis to the design so that physicians can better choose the most effective and cost-effective sequence of therapies for their patients. PMID- 11416937 TI - Identifying and treating rosacea. AB - Rosacea is a chronic, cutaneous vascular disorder that affects approximately 13 million Americans. The facial disorder, which often affects people of Northern European descent, is characterized by four stages. Although rosacea is not curable, early recognition and treatment can prevent progression to permanent disfigurement or blindness. Pharmacologic therapy, patient education, and symptom management can provide symptom control and remission. PMID- 11416938 TI - Recognizing hypochondriasis in primary care. AB - Between 5% and 9% of primary care patients exhibit hypochondriacal symptoms. Hypochondriasis may be primary or secondary to panic disorder or depression. Despite negative diagnostic findings and clinician reassurance, hypochondriacal patients manifest disease conviction, disease phobia, or both. Primary care providers, in collaboration with mental health care providers, can provide optimal treatment for patients with hypochondriasis. The use of various intervention phases and selective serotonergic reuptake inhibitors provide the most favorable patient outcomes. PMID- 11416939 TI - Determining the efficacy of glucosamine and chondroitin for osteoarthritis. AB - Glucosamine sulfate and chondroitin sulfate are being used by many patients for the treatment of osteoarthritis. Despite a number of studies supporting efficacy of these agents for palliation of joint pain in patients with osteoarthritis, the American College of Rheumatology Subcommittee on Osteoarthritis believes that it is too early to issue recommendations for use. Currently, the National Institute of Arthritis and Musculoskeletal and Skin Diseases in collaboration with the National Center for Complementary and Alternative Medicine have begun a pivotal study to thoroughly evaluate these agents. PMID- 11416940 TI - Lunelle: a new contraceptive alternative. PMID- 11416941 TI - The clinician-patient experience: understanding transference and countertransference. PMID- 11416942 TI - Hepatitis A virus seroprevalence in 1,000 university students in Toronto. PMID- 11416943 TI - Respiratory virus surveillance. FluWatch project, 2000-2001. End of season update. AB - The 2000-2001 season was a relatively mild season worldwide. In Canada, lower than usual activity was reported for all national indicators of influenza activity, including the rate of influenza-like illness (ILI), the percentage of laboratory-confirmed cases of influenza and provincial/territorial influenza activity levels. However, there were a number of interesting characteristics of this year's influenza season. In contrast to the predominance of influenza A, and in particular the A/Sydney/5/97 (H3N2)-like virus over the past 3 years, influenza B predominated overall this season. Influenza A (H3N2) accounted for < 1% of all characterized isolates (H1N1 accounted for 49% of isolates). Increased laboratory-confirmed influenza activity began in the West (Yukon, prairie provinces and British Columbia) in mid-December, followed by the Atlantic provinces in mid- to late January and Ontario and Quebec in mid- February and March. PMID- 11416944 TI - A reflective study of Alzheimer's caregivers. AB - Symbolic interaction posits that to truly understand a situation, it must be considered from both an observer's perspective and the perspective of the actor involved in the situation. In this study, the actors involved are former Alzheimer's caregivers (N = 20) and the situations examined are reflections of the caregiving experience. An in-depth interview was used to explore caregiver issues. The results show that former Alzheimer's caregivers remember their experiences vividly and can recount many stories regarding their successes, regrets, coping strategies, and barriers faced. Participants also shared how they coped with the death of their loved one, the major issues they had immediately following the death, and issues with which they are still dealing. From the information shared by former caregivers, an Alzheimer's Caregiver Transition Model (ACT-M) was developed to help explain the process individuals go through as they transition out of the caregiver's role. PMID- 11416945 TI - Correlation of rCBF (SPECT), CSF tau, and cognitive function in patients with dementia of the Alzheimer's type, other types of dementia, and control subjects. AB - BACKGROUND: The diagnosis of Alzheimer's disease (AD) during life remains difficult and a definite diagnosis of AD relies on histopathological confirmation at post-mortem or by cerebral biopsy. It is well known that levels of tau proteins are consistently and significantly increased in the cerebrospinal fluid (CSF) of Alzheimer's patients versus levels in normal controls. However, the sole use of this biochemical marker as a test for AD is hampered by mediocre specificity, since tau concentrations may also be elevated in certain other neurological disorders (OND). Studies of the regional cerebral blood flow (rCBF) are widely performed because of their convenience and usefulness in a variety of neurological disorders. Most studies have reported high diagnostic accuracy for brain perfusion single-photon emission tomography (SPECT) in Alzheimer's disease. METHODS: In order to improve specificity, in this study, correlation of 99mTc HMPAO SPECT scanning and CSF tau protein levels was made in 117 patients with AD, 67 patients with OND (26 of which had other dementias), and 23 age-matched controls. Means and standard deviations of tau protein levels were 297, 42 +/- 221, 12 in AD patients and 78, 07 +/- 98, 51 in patients with OND (p = 0.0006). No correlation was noted between CSF tau protein levels and age, duration of the disease, and neuropsychological scores of mini-mental state examination (MMSE), Cambridge Cognitive Examination (CAMCOG), and Functional Rating Scale for Symptoms of Dementia (FRSSD). FINDINGS: There was a bilateral parietal and temporal hypoperfusion in patients with AD in SPECT in comparison to normal subjects (p < 0.05) and there was a statistical correlation between this hypoperfusion and neuropsychological tests, such as MMSE and CAMCOG (p < 0.01). There was no correlation between tau protein levels and hypoperfusion in SPECT. INTERPRETATION: Conclusively, the correlation between elevated levels of tau proteins and hypoperfusion in SPECT in AD patients therefore cannot improve the specificity of tests in AD and this means that the determination of CSF tau proteins levels is not a specific diagnostic test for AD. PMID- 11416946 TI - Relinquishing in-home dementia care: difficulties and perceived helpfulness during the nursing home transition. AB - Institutionalization of an elderly relative suffering from dementia is a key transition in the caregiving career. This study determines factors that predict the problems and help dementia caregivers perceive while institutionalizing an elderly relative. By using a sample of 185 dementia caregivers interviewed before and after institutionalization, the authors conducted a multivariate regression to identify relevant predictors of search difficulties, professional help, and family help during the nursing home transition. The personal and social resources of caregivers prior to placement (e.g., mastery, family conflict, socioemotional support, duration of care, education, and physical health) were reliable predictors of search difficulties and perceived helpfulness. Such findings will prove useful to long-term care practitioners who wish to facilitate the institutionalization process for caregiving families. PMID- 11416947 TI - Products for Alzheimer's self-stimulatory wanderers. AB - The objective of this study was to develop a variety of sensory stimulation products for the behavioral intervention of patients with Alzheimer's type dementia. Many caregivers have relied on physical and chemical restraints as the primary method of patient intervention due to the lack of appropriate dementia management products. This significantly lowers the sufferer's quality of care and life. As the age group most susceptible to Alzheimer's disease (65 and older) is the fastest growing segment of our society, an appropriate care solution must be sought. The specific aim of this study was to develop products that are sensory satisfying for the Alzheimer's patient that exhibits self-stimulatory wandering behavior. Sensory satisfying objects for product development would be determined through structured observations of self-stimulatory wanderers in an institutionalized setting. Variations of product design and mounting would be pursued in order to develop products that are not only safe and effective for patient use, but are easy for the caregiver to implement and maintain. Such products would have widespread commercial application in both the institutional and private care settings such as nursing homes, adult day care facilities, Alzheimer's care facilities, convalescent homes, mental health institutions, and assisted-living facilities. PMID- 11416948 TI - Challenges of implementing intervention research in persons with dementia: example of a glider swing intervention. AB - Intervention studies for persons with dementia present many challenges. This article describes challenges encountered in conducting a glider swing intervention study for persons with dementia and the strategies used to manage the problems encountered. The overall purpose of a quasi-experimental study was to explore the effects of a glider swing intervention on emotions, relaxation, and aggressive behaviors in nursing home residents with dementia. Suggestions for investigators conducting intervention studies for persons with dementia are provided. PMID- 11416949 TI - AD vaccine shows promise in tests with genetically engineered mice. PMID- 11416950 TI - "Designer" gene may be effective against AD. PMID- 11416951 TI - Galantamine introduced in Europe. PMID- 11416952 TI - Human neural stem cells identified, isolated, and grown. PMID- 11416953 TI - Neotrofin clinical trials show promise. PMID- 11416954 TI - Vitamin E may help prevent AD for some people. PMID- 11416955 TI - Yale's HELP program prevents cognitive and functional decline in hospitalized elders. PMID- 11416956 TI - "Cognitive stress test" may yield early clues of preclinical AD. PMID- 11416957 TI - Cystitis glandularis mimics bladder cancer--three case reports and literature review. AB - Cystitis glandularis is an uncommon proliferative disorder of the mucus-producing glands within the mucosa and submucosa of urinary bladder epithelium. We report three cases with gross hematuria on first presentation. Both sonography and computer tomography revealing bladder tumor and transurethral resection was performed in all three cases. Cystitis glandularis was confirmed by pathology. Short-term follow-up of sonography and cystoscopy showed good results. PMID- 11416958 TI - Adult Wilms' tumor associated with polycythemia--a case report. AB - Both Wilms' tumor (nephroblastoma) in adults and polycythemia in Wilms' tumor are rare. Herein we report an extremely rare case of adult Wilms' tumor associated with polycythemia. A 41-year-old female was incidentally found to have right renal mass by abdominal sonography in a routine health examination. Laboratory examination revealed polycythemia (hemoglobin 20.2 g/dL). Although physical examination was unremarkable, CT scan revealed an homogeneous mass at the middle pole of right kidney, and chest x-ray revealed no metastatic lesions. Right radical nephrectomy was performed smoothly. Grossly, the tumor of 5 x 4.5 x 4.5 cm in size was well circumscribed, and had no vascular structure or collecting system involvement. Microscopic features were consistent with adult nephroblastoma. The post-operative course was uneventful. At follow-up, the patient was well, showed no evidence of recurrence and her hemoglobin level had returned to normal (hemoglobin 14.5 g/dl). We suggest that the relationship between polycythemia and Wilms' tumor should be carefully evaluated before surgical treatment. PMID- 11416959 TI - Endovascular embolization of carotid-cavernous fistula using the posterior communicating artery approach: a case report. AB - We report a case of direct carotid-cavernous fistula complicated with occlusion of internal carotid artery (ICA) at attempted endovascular obliteration via ICA and subsequent successful embolization via the posterior communicating artery (PCoA) approach. After a thorough understanding of vascular anatomical alterations associated with initial technical failure with detachable balloons, a microcatheter was navigated to the fistula site via the vertebro-basilar-PCoA to the distal cavernous segment of the left ICA. Embolization of the fistula was performed with deposition of three platinum microcoils. Angiograms after embolization showed complete disappearance of the fistula. There was no procedural complication and no recurrence of fistula at one-year clinical follow up. PMID- 11416960 TI - Expression of HER-2/NEU oncoprotein in familial and non-familial breast cancer. AB - The HER-2/neu proto-oncogene amplification or oncoprotein overexpression is an important prognostic factor and a predictive factor for resistance to endocrine therapy and adjuvant chemotherapy in breast cancers. Moreover, it is an entry criterion in the assessment of patients for whom Herceptin (Trastuzumab) treatment is considered. The overexpression rate of HER-2/neu oncoprotein has been identified in 10% to 40% of human breast cancers. In Taiwan, a higher grade of pathobiologic characteristics of familial breast cancer was also noted than that found in the non-familial group. It is worthwhile to evaluate whether the overexpression is more frequent in familial breast cancers. Fifty-six familial and 111 non-familial breast cancers were studied between 1990 and 1999 to assess both the overexpression of HER-2/neu oncoprotein immunohistochemically and the correlation with the histological type, grade and stage of breast carcinoma. The overexpression rate is higher in the familial breast cancer group (50.0%) when compared with non-familial breast cancer group (36.9%), which did not prove to be statistically significant (P = 0.1068). However, when the infiltrating ductal carcinomas of both groups are compared, it is statistically significant (52.3% vs. 33.7%, P = 0.0429). Overexpression correlated with node status and histological grade of infiltrating ductal carcinomas in non-familial and overall breast cancers. It also correlated with nuclear pleomorphism and mitotic counts, but not tubule formation or tumor size. All 3 cases of Paget's disease revealed overexpression, whereas all 12 cases of mucinous and one case of metaplastic carcinoma and one case of medullary carcinoma were negative. The overexpression rate was higher both in familial and non-familial intraductal carcinomas (57.1% vs. 73.3%, P = 0.4716). No statistical difference was identified between the 2 subsets. A case of infiltrating ductal carcinoma combined with intraductal carcinoma revealed heterogeneous staining in the component of ductal carcinoma in situ, while the invasive component did not. This suggests that overexpression decreases within individual tumors as they evolve from in situ to invasive lesioins. The HER-2/neu may imply a different role in intraductal carcinoma, Paget's disease and invasive duct carcinoma. Although the overexpression rate of HER-2/neu oncoprotein of familial breast cancer was not significantly higher than that of the non-familial group, it is appropriate to evaluate the rate of HER 2/neu overexpression according to the histological type of breast cancers from familial breast cancer and non-familial breast cancer. The prognoses will be needed for future evaluation. PMID- 11416961 TI - Effects of drugs on cellular proliferation in cultured iris pigment epithelial cells and retinal pigment epithelial cells. AB - In this study, iris and retinal pigment epithelial cells were cultured from porcine and various drugs including methionine-enkephalin, isoproterenol, dibutyryl cAMP, endothelin-1, dexamethasone and phorbol 12-myristate 13-acetate (PMA) were used to investigate their effects on both cellular proliferation in cultured porcine iris and retinal pigment epithelial cells. Cellular proliferation was estimated with 3H-thymidine uptake. It is indicated that both pigment epithelial cells possess epithelial-like morphology and abundant pigment granules in cells obviously. Following the iris pigment epithelial cells being treated with endothelin-1, the 3H-thymidine uptake in the cells was increased to 126% as compared with the control. However, the cellular proliferation was decreased to 83% when the cells were treated with isoproterenol. In the case of methionine-enkephalin, dibutyryl cAMP, dexamethasone and phorbol 12-myristate 13 acetate (PMA), the thymidine uptake in the iris pigment epithelial cells was not affected by above drugs. In the retinal pigment epithelial cells, the 3H thymidine uptakes were increased to 145% and 146% when the cells were incubated with methionine-enkephalin, and isoproterenol, respectively. In the presence of dibutyryl cAMP, dexamethasone and phorbol ester (PMA), the cellular proliferation was inhibited to 83%, 73% and 85% respectively. However, endothelin-1 did not affect the cellular proliferation in retinal pigment epithelial cells. These results show that the morphological shapes of iris pigment epithelial cells are similar to retinal pigment epithelial cells. However, the cellular proliferation in both cells may be regulated by distinct mechanisms. PMID- 11416962 TI - Cataract formation after pars plana vitrectomy. AB - To investigate the duration and pattern of cataract formation after pars plana vitrectomy, thirty-three eyes from 33 patients with complete data from March 1995 to January 2000 were collected in this study. The time and pattern of cataract formation was analyzed according to Lens Opacities Classification System III (LOCS III). The correlation between the mean cataract progression and follow-up time was determined by simple regression and correlation analysis. Diabetes mellitus and hypertension were the main causes of patients with vitreoretinopathy who underwent pars plana vitrectomy procedure. Twenty-one patients (63.6%) had diabetes mellitus and 11 patients (33.3%) had hypertension. Four eyes (13.8%) received intravitreal gas injection after pars plana vitrectomy. The correlation between cataract progression and follow-up time was clinically significant (p < 0.05). Thirty-one patients (94%) were nuclear cataract, the mean time of cataract formation being 9.1 months; 15 patients (46.9%) were cortical cataract, the mean time of formation being 8 months; and 24 patients (72.7%) were posterior subcapsular cataract, the mean time of formation being 13.3 months. Cataract formation after pars plana vitrectomy is not infrequent. Nuclear cataract is the most common type in this study. Vitreous microenvironmental changes, post vitrectomy uveitis, intravitreal gas, and patient age may be the key points of cataract formation after surgery. The mechanism of longer-term effect of pars plana vitrectomy on lens status is still unknown and merits further study. PMID- 11416963 TI - Classifiers and social control in wheelchair rugby. AB - The purpose of this study was to examine the features of wheelchair rugby classifiers in order to maintain social control in wheelchair rugby. The examination centered on the theme-resources used by classifiers to maintain the authority of International Wheelchair Rugby Federation (IWRF). Data were collected using participant observation methodology at national and international wheelchair rugby championships and a survey of the 14 IWRF authorized classifiers. The results of this study identified five essential features of IWRF classifiers. In particular, they need to have a background in medicine or physical therapy. In addition, IWRF classifiers use their medical and sporting knowledge and experience to control the wheelchair rugby classification process and to maintain fairness of competition. The results of this study may help to train Taiwanese professionals with medical or sporting backgrounds to become international classifiers and also to develop appropriate training programs for national classifiers. PMID- 11416964 TI - Acute open perilunate dislocation--a case report. AB - A rare case of acute open dorsal perilunate dislocation of the left hand was encountered in a 30-year-old man with exposure of the lunate and distal radius. No other combined injury was identified except open dislocation and chip avulsion fracture of triquetrum. Open reduction was performed through a dorsal approach after debridement. Such a severe injury, to our knowledge, has not been reported previously. PMID- 11416965 TI - Symptomatic mechanical heart valve thrombosis: high morbidity and mortality despite successful treatment options. AB - BACKGROUND: Recommendations for treatment of mechanical prosthetic heart valve thrombosis (PVT) include systemic thrombolysis and/or reoperation. Data on complications and outcome are limited. METHODS: Clinical and echocardiographic findings of 17 patients with mechanical PVT were reviewed. Complications and outcome of surgery and/or thrombolysis were analysed. Prospective follow-up was obtained. RESULTS: Symptomatic PVT occurred 8.4 +/- 7.2 years after mechanical valve replacement at mean age 55 +/- 15 years. Thrombosis involved the mitral valve in 12 patients (71%), the aortic valve in 4 (24%) and the tricuspid valve in one (6%). The reason for PVT was inadequate anticoagulation in 11 patients (65%), endomyocardial fibrosis in 2 (12%) and unknown in 4 (24%). Prior to diagnosis, systemic emboli occurred in 6 patients (35%). Thirteen patients (76%) presented in functional class NYHA IV. Haemodynamic valve obstruction was documented by echocardiography in 15 patients (88%). Treatment included primary reoperation in 12 patients (71%), thrombolysis with urokinase in 3 (18%) (with reoperation in 1), reinstitution of adequate anticoagulation in one (6%); death occurred before treatment in one (6%). Intraoperatively, both pannus and thrombus were found in 5 of 13 patients (38%). Treatment-related emboli occurred in 5 patients (29%), to the brain in 3, to the legs in one and to a coronary artery in one. Five patients died (mortality 29%) within 30 days due to multiorgan failure/septicaemia (3 patients), congestive heart failure (1), or cerebral emboli (1). Follow-up after 28 +/- 28 months in the 12 surviving patients was unremarkable. CONCLUSIONS: The most common aetiology for obstructive PVT is thrombus formation due to inadequate anticoagulation. PVT remains a serious complication with high morbidity and mortality despite aggressive treatment by thrombolysis and/or surgery. Surgery is often needed due to the frequent presence of pannus and/or large thrombi. However, long-term prognosis after successful treatment of PVT is excellent. PMID- 11416966 TI - Renal transplantation with donors aged over 50: a long-term, single centre experience. AB - BACKGROUND: Renal transplantation is now considered the best treatment for end stage renal failure patients. In view of the worldwide growing waiting list, extending donor age is one of the strategies implemented to make more kidneys available for transplantation. METHODS: From October 1983 to February 2000, 324 renal transplantations were performed at Geneva University Hospital, 68 of which were done with kidneys from donors aged over 50 (mean donor age 55.3 + 4.8 years) The following data were analysed at 1, 5 and 10 years and compared between recipients with donors aged over 50 (> 50 years) and under 50 (< 50 years): actuarial patient and graft survival, serum creatinine, causes of graft loss. RESULTS: Patient survival at 10 years was 83% with donors < 50 years and 61% with donors > 50 years (non-significant). Graft survival at 10 years was 61% with donors < 50 years and 48% with donors > 50 years (non-significant). Graft survival at 10 years or death with functioning graft was 69% with donors < 50 years and 68% with donors > 50 years (non-significant). Overall, donor age was not a predictive factor for graft survival, as shown by multiple logistic regression. CONCLUSIONS: Kidneys obtained from donors > 50 years are suitable for renal transplantation and yield acceptable graft survival. PMID- 11416967 TI - Macroprolactinaemia, the major unknown in the differential diagnosis of hyperprolactinaemia. AB - We report on 10 cases of macroprolactinaemia and discuss recent evidence that many patients with hyperprolactinaemia (8-26%, depending on the population studied) have in fact normal amounts of circulating prolactin but false-high values in commercial assays. This is caused by macromolecular prolactin (also named big-big prolactin or macroprolactin), a complex of prolactin with IgG antibodies leading to apparent hyperprolactinaemia. In spite of the expanding literature on this topic, it remains an underrecognised problem, typically causing unnecessary procedures such as laboratory controls, MRI of the pituitary, treatment with dopamine agonists or even pituitary surgery. Physicians involved with diagnosis and treatment of hyperprolactinaemia (general practitioners, gynaecologists, neurosurgeons, endocrinologists and biochemists) should suspect the presence of apparent hyperprolactinaemia in any patient with a high prolactin value but no related symptoms. Medical laboratories should be aware that their prolactin assay can interfere with macroprolactin and should implement the use of the PEG precipitation test in the work-up of hyperprolactinaemia, a simple and effective means of correctly diagnosing apparent hyperprolactinaemia. PMID- 11416968 TI - Metabolic and endocrine effects of metabolic acidosis in humans. AB - Metabolic acidosis is an important acid-base disturbance in humans. It is characterised by a primary decrease in body bicarbonate stores and is known to induce multiple endocrine and metabolic alterations. Metabolic acidosis induces nitrogen wasting and, in humans, depresses protein metabolism. The acidosis induced alterations in various endocrine systems include decreases in IGF-1 levels due to peripheral growth hormone insensitivity, a mild form of primary hypothyroidism and hyperglucocorticoidism. Metabolic acidosis induces a negative calcium balance (resorption from bone) with hypercalciuria and a propensity to develop kidney stones. Metabolic acidosis also results in hypophosphataemia due to renal phosphate wasting. Negative calcium balance and phosphate depletion combine to induce a metabolic bone disease that exhibits features of both osteoporosis and osteomalacia. In humans at least, 1,25-(OH)2 vitamin D levels increase, probably through phosphate depletion-induced stimulation of 1-alpha hydroxylase. The production rate of 1,25-(OH)2 vitamin D is thus stimulated, and parathyroid hormone decreases secondarily. There is experimental evidence to support the notion that even mild degrees of acidosis, such as that occurring by ingestion of a high animal protein diet, induces some of these metabolic and endocrine effects. The possible role of diet-induced acid loads in nephrolithiasis, age-related loss of lean body mass and osteoporosis is discussed. PMID- 11416969 TI - Thalidomide: near complete regression of extramedullary bulk in refractory multiple myeloma. PMID- 11416970 TI - [Your ultrasound diagnosis? Key ring 2.5 cm in diameter, freely mobile in the stomach]. PMID- 11416972 TI - [Hyper- and hypothyroidism: which tests are reliable in general practice?]. AB - Thyroid function is evaluated by measuring the serum concentrations of thyroid stimulating hormone (TSH) and free thyroxine (fT4). To confirm a normal thyroid function, a TSH within the normal range is usually sufficient. If hyperthyroidism or hypothyroidism is suspected clinically, the assessment of TSH in association with fT4 is recommended. Additional tests in the evaluation of hyper- and hypothyroidism should be performed depending on the clinically suspected diagnosis. Acquired primary hypothyroidism in the adult can be diagnosed by measurement of TSH and fT4. Additional investigations are usually unnecessary. The indications to perform a thyroid radionuclide scan, thyroid ultrasonography and the determination of thyroid antibodies in patients with primary hyperthyroidism are discussed. PMID- 11416973 TI - [Endocrine orbitopathy]. AB - Dysthyroid orbitopathy is an autoimmune disorder usually occurring in the setting of Graves' disease. The pathogenesis is incompletely understood but involves autoantibodies against orbital tissue. The clinical course is highly variable. Signs range from mild to severe soft tissue swelling, lid retraction, proptosis and extraocular muscle involvement with diplopia, to corneal damage and optic nerve compression with irreversible visual loss. After a few years, the phase of active inflammation is usually followed by a chronic stage with or without residual damage through fibrosis. Each stage requires a carefully adjusted therapy and optimal cooperation between the general practitioner, the ophthalmologist and other specialists involved. Therapeutic options range from topical lubricants to systemic steroids, radiation and surgery. PMID- 11416974 TI - [Is the general practice laboratory medically and economically valuable?]. AB - Between 1998 and 2000 we evaluated the office-based laboratory activities of general practitioners. The aim was to clarify whether there is a medical and economic benefit of these activities. METHODS: The study was performed in four parts: I. A cross-sectional study with a random sample of general practitioners of the German and French speaking part of Switzerland. II. A prospective evaluation of the office-based laboratory activities of 56 GP's. III. A cross sectional study of the preference of 837 patients in 52 of GP's offices. IV. A consensus panel with nine experts using the RAND method. RESULTS: 1999 there were 55.4 Million laboratory tests ordered by GP's (excl. pediaters) of which 78.9% were analysed in the office-based laboratory. The probability of a second visit is reduced by 60%, if all of the tests could be performed in the office-based laboratory. 85% of the patients appreciate the possibility to discuss the test results within the same consulation. In the consensus panel, 43 tests were proposed of which only bicarbonate, chloride and urea were assessed as not useful for the office based laboratory. CONCLUSION: The office-based laboratory is a well embodied institution in Switzerland. It's predominant advantage is the possibility of point of care testing. It allows a quick management of the patient and avoids unnecessary second consultations. PMID- 11416975 TI - [Current epidemiology and therapy of renal failure]. AB - The worldwide increase of chronic renal failure is a challenge for cost effective therapeutic modalities of high quality. It affords timely communication and cooperation between practitioners and specialists. PMID- 11416976 TI - [Quality development of the internet. A pilot project]. PMID- 11416977 TI - [Seroepidemiology of toxoplasmosis in a marginal community of the Municipality of Maracaibo, Zulia State, Venezuela]. AB - The goal of this study was to determine the prevalence of anti-Toxoplasma gondii antibodies in a marginal community of the Municipality of Maracaibo, Zulia State, Venezuela and if there was a relationship between risk factors and infection. An epidemiological survey of personal data, living sanitary conditions and contact with cats was performed. A collection of 254 blood samples from male and female individuals, with ages ranging from 8 months to 76 years, was studied. Anti Toxoplasma gondii reaction was determined by indirect hemaglutination using a commercial kit. A positive reaction at a dilution > or = 1:64 was considered positive. Statistical analysis was performed using the Chi-squared test. The prevalence of infection was found to be 36.6%. It was observed that 21.5% of the seropositive presented titers not considered significant (< or = 1:64), 51.6% had intermediate titers (1:128 to 1:512), 24.8% high titers (1:1024 to 1:4096) and 2.1% very high titers (> or = 1:8192). The highest percentage of positivity (50%) was observed in the group > or = 46 years of age. No statistically significant association was observed between prevalence and risk factors (living together with cats and poor sanitary conditions of living) or infection when the chi square test was used. Toxoplasma gondii infection was often found in the community of Puerto Caballo and probably cats and poor living conditions have a role in the transmission of infection. PMID- 11416978 TI - [Concentration of thyrotropic hormone and free thyroxin in children with Down's syndrome]. AB - The incidence of hypothyroidism is higher among children with Down syndrome than among children in the general population. The frequency of hypothyroidism is higher in the areas of endemic goiter than in other areas. The aim of this paper was to study the concentrations of TSH and FT4 in children with Down syndrome residents of Merida, a region of Venezuelan Andes. At the Centro de Estudio y Prevencion del Retardo Mental y Alteraciones en el Desarrollo (CEPREMAD), the thyroid function was studied in 48 children (1 month to 6 years old), who had Down syndrome, and in 123 healthy children of similar ages. All the children were referred to the Center for thyroid function evaluation. Two (4.2%) of the 48 children with Down syndrome had congenital hypothyroidism and 22 (45.8%) had subclinical hypothyroidism (high concentration of thyrotropin-TSH). Among the control children, only 14% had elevated levels of TSH. There were no differences in relation to the gender. In conclusion in children with Down syndrome, the frequency of high concentrations of TSH was three times higher than the frequency among the healthy children. The frequency of hypothyroidism was similar to that found in areas without endemic goiter. PMID- 11416979 TI - [Effectiveness of Bacillus sphaericus strain 2362 on larvae of Anopheles nuneztovari]. AB - A study, under laboratory and field conditions, was conducted to evaluate the effectiveness, persistence in the time and some factors that can affect the larvicidal action of the Griselef formulation of the sporogenous bacteria Bacillus sphaericus strain 2362, on the larvae population of the Anopheles nuneztovari Gabaldon, the main malaria vector in western Venezuela. The degree of susceptibility of this species to the Bacillus sphaericus was determined to be at a LD50 in 0.07 ppm a LD95 in 0.69 ppm. The pH ranges of the water, as tested in the laboratory, showed that values between 7.0 and 9.0 did not affect the action of the larvicide, producing more than 95% of mortality after 72 hours of observation. Under controlled field conditions, the high susceptibility of the An. nuneztovari larvae to the larvicidal action of the product was demonstrated. With a dose of 5 ml/m2, a 100% mortality was obtained after 72 hours of exposure. In the field, it was determined that after 12 hours, solar radiation decreased the effectiveness of the B. sphaericus in 30% and that the presence of vegetation reduced the biolarvicidal effect between 5 and 12%. Under natural field conditions, it was shown that the larvicide was effective, since a 100% larval mortality was obtained in the two treated breeding places. This effect persisted up to 4 months of observation, with more than 85% of larvae reduction, showing that it is capable or remaining in the natural environment. PMID- 11416980 TI - [Giant fibrolipoma of the floor of the mouth. Presentation of a clinical case]. AB - The fibrolipoma is a benign tumor variant of the lipoma, characterized by the presence of adipose and fibrous tissues. The authors report a case of a big oral fibrolipoma in a 72 year old woman. After surgery, a mass of 13 x 8 x 6 cm was obtained. The tumor had an implantation pedicle of 1 cm, on the floor of the mouth. The microscopic evaluation showed the presence of polygonal cells grouped into nests and separated by fibrous connective tissue septa. We have not found any report in the literature related to a fibrolipoma located on the floor of the mouth with the characteristics presented in this work. PMID- 11416981 TI - [Importance of the different species and genotypes of Cryptosporidium in public health]. AB - Until recently, only the bovine and human genotypes of Cryptosporidium parvum had been considered to infect man and to be potential pathogens. Lately, symptomatic infections with C. felis, C. meleagridis, and C. parvum-like dog genotype have been described in immunocompetent and immunosuppressed individuals. These findings suggest that zoonotic parasites could also play an important role in Public Health. PMID- 11416982 TI - [Liver function of workers occupationally exposed to mixed organic solvents in a petrochemical industry]. AB - A descriptive and cross sectional study was conducted to determine whether hepatic function changes in workers occupationally exposed to a mixture of organic solvents, were due to the exposure or confusing factors. A non random sample of 77 workers, operators and supervisors of the Olefin Plant I and II of a petrochemical industry in Maracaibo, Venezuela, was used. Their mean age was 29 +/- 7 years, and had at least one year of exposure to the solvents. This sample was compared with a group of employees of the administrative offices or control panel workers, with a mean age of 36 +/- 8 year and with similar anthropometric characteristics. Workers with a known history of liver disease, blood transfusions and diabetes mellitus were excluded of the study. In addition to a complete occupational disease medical history and a physical examination, serum samples were obtained to determine the activity of the aspartato aminotransferase (AST), alanin aminotransferase (ALT), gamma glutamiltransferase (GGT), alkaline phosphatase (AF), the concentration of the total bile acids (BAS), the surface antigen of hepatitis B(HbsAg) and the hepatitis A virus antibodies: AntiHAV-IgG and the AntiHAV-IgM. An urine sample was taken and analyzed by standard methodology to determine urinary phenols. The air concentrations of benzene, ethylbenzene, toluene and xylene were analyzed by gas chromathography. The serum activities of the liver enzymes, the concentration of bile acids and urinary phenols were not influenced by the exposure to the solvents. The increase of the activity of GGT was associated with obesity and alcohol consumption. The antibodies of the surface antigen of hepatitis A-IgM were normal in both groups and the antibodies for the antigen of hepatitis A-IgG presented a prevalence of 6% in the exposed group and 9% in the non exposed not being associated with liver abnormalities. The individual air concentrations of the solvents were below the environmentally permissible concentrations, except one sample of benzene (1, 14 ppm) that was over the allowed limit. The total maximum concentration of the mixture of organic solvents, resulting of the sum of fractions of each organic solvent, was within the allowed limits. In conclusion, obesity and alcohol consumption, and not the occupational factors, seem to be responsible for the alteration in GGT in workers of these Olefin Plants. PMID- 11416983 TI - [Human microsporidiosis]. AB - This review shows the Microsporidia as unicellular protozoa strictly intracellular eukaryotic parasites of animals and humans. These study concerns life cycles, cytology, host-parasite relationships in animal models experimentally infected with microsporidia from human feces, demonstrating the host-inespecificity and visceral dissemination with histopathological studies in digestive, tract, kidney, liver, spleen, brain, heart, pancreas, thyroid, suprarenal glands. It is presented the microsporidiosis in Venezuela in immunocompetent patients and immunodeficients HIV+, with diarrheic syndromes with keratoconjuntivitis microsporidial punctata diffuse and with disseminated microsporidia in urine, tracheobronchial sputum, nasal and pharyngeal exudates. Also we have found the microsporidia in river and lake waters and in animals relationed with the man: dogs, cats, pigs, monkeys, donkeys, guts. The patients are treated specially with Albendazole and also with Trimethoprim-Sulphametoxazol for the children until two years of age. We use as laboratory diagnosis the technique of Kinyoun stain, in support of the acid resistance property of these parasites, and also the Chromotrope staining. Our recommendations for a proper identification of the Microspora species, should be done with Electron Microscopy and the TCR reaction. PMID- 11416984 TI - [Why are plasmatic apoB lipoproteins atherogenic? The hypothesis of response to retention]. AB - The development of atherosclerotic lesions can be described as a tissular response to deposition of apoB-100 containing lipoproteins (LpApoB) in the arterial intima. These particles that circulate in blood are the low-density lipoproteins (LDL), the very low and intermediate density lipoproteins (VLDL and IDL) and the Lp(a). To initiate the tissue response is critic that LpApoB are retained in the subendothelial space. This occurs by the interaction of specific positive segments of the apoB-100 with negative glycosaminoglycan chains of the proteoglycans of the intima extracellular matrix. The inflammatory response involve macrophages and other immuno-competent cells, smooth muscle cells and endothelial cells. The direct agents that induce this reaction appear to be products of hydrolytic and oxidative modifications of the phospholipids, triglycerides and sterols of the retained LpApoB. The enzymes that cause these modifications are secreted by arterial cells stimulated by pro-inflammatory cytokines. The sequence of LpApoB retention and modification in the extracellular matrix followed by proliferation and inflammation seems to be a cyclic process that leads to the chronic progress of atherosclerotic lesions. These concepts constitute the base of a new hypothesis to explain atherosclerosis: the "response to retention hypothesis". PMID- 11416985 TI - Thrombotic thrombocytopenic purpura--a syndrome caused by multiple pathogenetic mechanisms. AB - Platelet thrombus formation in small vessels in thrombotic thrombocytopenic purpura (TTP) is triggered by certain stimuli to cause vascular injury, primary platelet aggregation, or both. The formation and dissolution of platelet thrombi are modulated by proteolysis, plasma factors, PGI2 synthesis and stability, and immune mechanisms. It has been demonstrated that TTP plasmas contain heterogeneous platelet aggregating factors and that TTP plasmas induce the apoptosis of microvascular endothelial cell. Fibrinolysis has been shown decreased. Von Willebrand factor (vWF) under high shear stress is unfolded and becomes adhesive to platelets to induce platelet aggregation. Recently it is found that vWF protease is deficient in hereditary TTP, intermittent relapsing TTP, idiopathic acute TTP and ticlopidine- and clopidogrel-induced TTP, but normal in hemolytic uremic syndrome and organ transplantation-related thrombotic microangiopathy. Apparently absence of vWF protease leaves unfolded adhesive vWF unchecked to cause platelet aggregation. Antibodies against vWF protease are present in intermittent relapsing TTP, idiopathic acute TTP, and ticlopidine induced TTP. Plasma exchange and/or infusion with or without corticosteroids remain the cornerstone of treatment. PMID- 11416986 TI - Effect of processing methods and heat treatment on the formation of wax matrix tablets for sustained drug release. AB - The objective of this study was to evaluate the effects of processing methods and heat treatment on matrix formation and subsequent drug release from wax matrix tablets for controlled release. Phenylpropanolamine hydrochloride (PPA) and Compritol were processed with appropriate diluent(s) using either dry blending (DB), wet granulation (WG), partial melt granulation (PMG), or melt granulation (MG). Then the tablets were heat-treated at 80 degrees C. Particle size distribution and compressibility, along with drug release, tablet micro morphology, wettability, porosity, and tortuosity were investigated. The drug release was different for the four processing methods even though the tablet formulation was identical. Heat treatment further retarded drug release and its effect was related to the previous manufacturing processes. Scanning Electron Microscopy (SEM) showed that heat treatment redistributed the wax and formed a film-like structure covering drug and excipients. The contact angle of tablets made from DB, WG, and PMG methods increased after heat treatment, while that of tablets made from MG remained constant. Tablet tortuosity calculated from drug release rate constants increased dramatically after heat treatment. Drug release from the wax tablets with or without heat treatment was best described by the Higuchi equation. Different processing methods produced different matrix structures that resulted in different drug release rates. Heat treatment retarded drug release mainly by increasing tortuosity of the matrix. Contact angle measurement and SEM analysis indicated that heat treatment caused the wax to melt, redistribute, coat the drug and diluents, and form a network structure. PMID- 11416987 TI - Solubilization and stability of bumetanide in vehicles for intranasal administration, a pilot study. AB - The solubility of bumetanide in vehicles of various polarities, suitable for intranasal administration in acute situations, has been investigated. The solubility at 4 degrees C in glycofurol and polyethylene glycol 200 was high (167 and 143 mg/mL, respectively), decreasing exponentially with addition of phosphate buffer or coconut oil. Vehicles containing coconut oil and glycofurol did not seem to improve the solubility relative to mixtures between glycofurol and buffer. Adequate solubility (approximately 50 mg/mL) was achieved in vehicles containing about 80% cosolvent. The stability of bumetanide was studied at 5 degrees C and 57 degrees C. No degradation was observed at low temperature. At high temperature, bumetanide decomposes in nonaqueous vehicles with half-lifes ranging from 69 to 400 days, but sufficient stability may be obtained by adjustment of pH to 7.4. It may be concluded that it is possible to prepare a clinically relevant formulation for intranasal delivery of bumetanide. PMID- 11416988 TI - Freeze drying properties of some oligonucleotides. PMID- 11416989 TI - The influence of excipients on the stability of the moisture sensitive drugs aspirin and niacinamide: comparison of tablets containing lactose monohydrate with tablets containing anhydrous lactose. AB - The purpose of this study is to test the hypothesis that in tablet formulations, moisture-sensitive drugs formulated with lactose monohydrate have the same stability as formulations containing anhydrous lactose, and to characterize the kinetics of niacinamide degradation in the solid state. Aspirin and niacinamide decomposition were used as indicators of stability. Aspirin and niacinamide tablets containing either lactose monohydrate or anhydrous lactose were separately investigated at different temperatures and relative humidities; the stability tests were done at 25 degrees C--60% RH, 40 degrees C--80% RH, 60 degrees C--60% RH, 60 degrees C--80% RH, and 80 degrees C--80% RH. Official U.S. Pharmacopeia methods were used for the aspirin and niacinamide assays. Statistical analysis showed that tablets containing lactose monohydrate have the same stability as tablets containing anhydrous lactose, which means that even though water is present in the crystal structure, the bound water does not influence the reaction rate. In addition, niacinamide degradation in the solid state can be described by a third order rate equation. PMID- 11416990 TI - A novel approach to increase oral drug absorption. AB - A comprehensive analytical solution that accounts for many factors and assumptions affecting drug concentration profile in the gastrointestinal tract was presented. A sensitivity analysis approach was utilized in order to investigate the importance of different parameters in the model. The partition coefficient is found to be the key parameter. Hence, for drugs stable in the intestinal wall, increasing partition coefficient only can lead to higher drug absorption. However, for drugs unstable in the intestinal wall, increasing partition coefficient and diffusivity is needed to counteract drug instability. On the other hand, the model is essentially insensitive to degradation in the intestinal lumen for degradation half lives greater than 0.7 min at a given intestinal length. However, a high model sensitivity to the rate of luminal degradation is obtained at higher intestinal length values. The proposed model can be used as a guide for oral drug delivery. PMID- 11416991 TI - Release behavior of ketoprofen from poly(acryloyl-L-proline methyl ester) gels having different crosslinked networks. AB - In order to clarify the relationship between the crosslinked structure of thermo responsive polymer and drug release profile, polymer gels based on acryloyl-L proline methyl ester (A-ProOMe) were synthesized in a mixture of water and acetone by the following two methods: a simultaneously occurring process of radiation-induced polymerization and crosslinking without a crosslinker (self bridging method), and radiation-induced polymerization in the presence of the crosslinker tetradecaethylene glycol dimethacrylate (crosslinker method). The pronounced gap in thermo-response between two A-ProOMe gels, with an apparent degree of crosslinking of 11 for 1-propanol, shows a different shrinking pattern in the initial stage of time. The gels, which were obtained with the self bridging method and the crosslinker method, were kept constant at a swelling ratio of 17 in water at 0 degree C for all systems. However, those values fell to 0.5 and 4, respectively, at 10 min after the temperature was increased to 37 degrees C. The release mechanism of ketoprofen from two gel devices showed an anomalous (non-Fickian) transport, in which the release of ketoprofen with a low water-solubility could be directly related to the rapid release of water accompanying a gel shrinkage. PMID- 11416992 TI - End-point detection in a wet granulation process. AB - The purpose of the experiments in this paper was to evaluate granulation end point control by power consumption measurement in a 25-L high shear mixer. The effect of impeller design, impeller speed, liquid addition rate, type of binder, and mixing ratio between lactose and starch on the correlation between power consumption and granule growth was investigated in the liquid addition phase of the process using a fractional factorial experimental design. There was found generally a linear correlation between power consumption and mean granule size, but this correlation was dependent on the impeller design, the impeller speed, and the type of binder. However, an end-point control based on power consumption was not found to be sensitive to variations in the lactose:starch ratio. It was concluded that it was possible to control the liquid addition by the level detection method by which the liquid addition is stopped at a predetermined level of power consumption. An end-point control based upon the peak detection method was not generally applicable, because a peak in the differentiated power consumption signal could not be identified in all the experiments. PMID- 11416993 TI - Influence of elastic deformation of particles on Heckel analysis. AB - The Heckel equation is one of the most useful equations for describing the compaction properties of pharmaceutical powders. Important material properties (e.g., yield strength) of powders can be derived using Heckel analysis. Two types of Heckel analysis are in common use. One is the "out-of-die," or "zero-pressure" method, the other is the "in-die" or "at-pressure" method. Because particles undergo elastic deformation under pressure, which tends to lower the porosity of the powder bed, the "out-of-die" method describes powder consolidation and compaction more accurately than the "in-die" method. However, "in-die" Heckel analysis has been widely used because of the speed and ease of data collection. Using L-lysine monohydrochloride dihydrate as a model compound, this work analyzes quantitatively the effects of elastic deformation on the calculation of porosity of a tablet, and therefore on the Heckel analysis. The effects of a small change in porosity, epsilon, on Heckel analysis are presented mathematically. It is found that a decrease in porosity of 0.001, when the porosity is lower than 0.05, causes a significant increase in the value of -ln epsilon. Therefore, data at epsilon < 0.05 should be interpreted with caution when using Heckel analysis. Elastic deformation causes positive deviations in the Heckel plot, and therefore leads to a yield strength that is lower than the true value. The lower the elastic modulus of the powder, the greater is the deviation from the true value. Therefore, the "in-die" method gives values of yield strength that are significantly lower than the true values for most pharmaceutical powders. PMID- 11416994 TI - Effervescent atomization of aqueous polymer solutions and dispersions. AB - The purpose of this paper is to characterize the performance of effervescent atomizers via the effects of Air-to-Liquid-Mass-Ratio, (ALMR), feed pressure, and liquid viscosity on spray droplet size. Water and aqueous solutions having varying concentrations of polyvinylpyrrolidone, (PVP), (Kollidon 30, 90, and the copolymer VA 64), hydroxypropyl methylcellulose, (HPMC), (Pharmacoat 603, 606, and 615), and aqueous polymethacrylate dispersions, (Eudragit RS 30 D, RL 30 D, and L 30 D-55) were sprayed over a range of ALMRs from approximately 0.1 to 0.8 at feed pressures ranging from approximately 550 to 1590 kPa. Fluid viscosities ranged from 1 to 228 mPa.s. The ALMR of the spray was shown to be the most important factor influencing the Sauter mean diameter (SMD) of the droplets, while feed pressure was shown to have only a minor effect on droplet size. Sprays with SMDs between 7 and 12 microns were produced for the polymethacrylates, 13 to 24 microns for the PVPs, and 15 to 20 microns for the HPMCs, all at an ALMR of 0.4 and a feed pressure of 1172 kPa. A higher liquid viscosity lead to larger droplets. The correlation between viscosity and droplet size was found to be dependent on the type of polymer. PMID- 11416995 TI - Influence of film additives on stabilizing drug release rates from pellets coated with acrylic polymers. AB - The objective of this study was to investigate the influence of talc and triethyl citrate (TEC) on stabilizing the drug release rates following curing and storage at elevated temperature of pellets coated with an aqueous acrylic polymeric dispersion. Core pellets containing anhydrous theophylline (20%), microcrystalline cellulose, and polyvinylpyrrolidone were prepared by extrusion spheronization. The aqueous dispersions were prepared by adding up to 30% TEC as a plasticizer and talc up to 200% as an antiadherent to a mixture of Eudragit RS 30D/RL 30D (95:5). The theophylline pellets were coated in a fluidized-bed coating unit and then cured at elevated temperatures. Theophylline pellets were successfully coated with the Eudragit dispersions that contained up to 200% talc, based on the dry polymer weight, and the coating efficiency was greater than 93%. Our results demonstrated that the polymer, which was plasticized by TEC, was able to function as a film-forming agent for dispersions containing high levels of talc. No sticking of the coated pellets was observed during the coating process or during the curing or equilibrating phase, even with high levels of TEC in the film. The dissolution rate of theophylline from the coated pellets was delayed when the film coating dispersion contained high levels of talc. Additionally, the stability of the drug release profiles from the coated pellets after storage was significantly improved. Furthermore, a modified dissolution testing used to simulate mechanical stresses that may be encountered in vivo showed the film coated pellets would have sufficient strength. The results of this study demonstrated that high levels of film additives in the acrylic dispersion contributed to the stabilization of the drug release rates as well as the reproducibility of the coating process. PMID- 11416996 TI - Stability of extruded 17 beta-estradiol solid dispersions. AB - Recrystallization is one of the main problems concerning the stability of solid dispersions. Different analytical methods were applied showing that no recrystallization occurred after treating melt extruded solid dispersions with 17 beta-Estradiol as the model drug with heat or water vapor. A skillful choice of excipients--a combination of polymers and additives--could be the reason for improving the stability. The requirements of the USP 23 for Estradiol tablets of 75% dissolved drug after 60 min were fulfilled after storing the tablets for 6 months at 40 degrees C/75% RH. By observing the change in glass transition temperature, DSC analysis showed that the solid dispersions were stable against thermal stress. Isothermal microcalorimetry as well as moisture absorption gravimetry were methods to prove the stability of the solid dispersions against water vapor. PMID- 11416997 TI - Preparation and release characteristics of protein-loaded polyanion/gelatin complex. AB - PURPOSE: This paper describes preparation of polymethacrylic acid/gelatin complex and Myoglobin release characteristics in order to evaluate the polyanion/gelatin complexes as matrices that can release proteins at a near zero-order kinetics over a long period of time. METHODS: Mb-loaded PMAA/gelatin complex was prepared by two different titration methods. Mb entrapment efficiency and PMAA/gelatin ratio in the complex were determined by HPLC. The release of Mb and gelatin from the complex was followed by HPLC. Mb conformation was detected by UV-vis spectrophotometer and capillary electrophoresis apparatus. RESULTS: Polyanion/gelatin feed ratio of the polyanion/gelatin/Mb mixed solution has great effects on complex yield and protein entrapment efficiency when "Type I" titration method is adopted, while for the colloid titration method the complex yield and protein entrapment efficiency are hardly influenced by preparative conditions (ca. 100%). Mb release rate could be adjusted by the complex composition (e.g., PMAA MW, hydrophobilization of PMAA, Mb loading and PMAA/gelatin ratio, etc.). Moreover, by coating of high MW PMAA/gelatin complex cylinder in a hydrophobic membrane with one open-end left, the period of protein release can extend to ca. 20 days and the release displays a near zero-order pattern. The protein release profiles can be described by the dissociation/erosion mechanism. The entrapment process has little effect on Mb conformation. CONCLUSIONS: The studied polyanion/gelatin complex is promising to be used as protein carriers to release proteins at a near zero-order kinetics over a long period of time by selecting suitable polyanions and designing the device structure. PMID- 11416998 TI - Monitoring the retention of a protein antigen in complete Freund's adjuvant, alum, and pluronic F-127 gel formulations by X-ray fluorescence. AB - Adjuvants function by protecting antigens from rapid degradation or dispersal. The effectiveness of experimental adjuvants can be assessed by measuring antibody titers to the antigen of interest or, less frequently, by evaluating the retention and distribution of antigen at the application site. In this study, we used X-ray fluorescence (XRF) to monitor the release of an iodinated protein (I bovine serum albumin) from several adjuvant formulations after its subcutaneous injection in rats. The interaction of the tagged antigen with an external Am-241 source leads to the emission of iodine X-rays from the application site; the number of these X-rays is proportional to the concentration of the protein remaining at the injection site. The disappearance of the iodine X-rays, and hence the antigen, from the injection site followed first-order kinetics for all adjuvant formulations tested; mean half-life values were as follows: in 50% Freund's adjuvant, 17.1 +/- 1.1 h; in 4-hour-old 25% Alum, 11.78 +/- 0.08 h; in 4 h-old 50% Alum, 13.2 +/- 2 h; in 3-day-old 50% Alum, 15.8 +/- 1.5 h; and in 240 mg/mL Pluronic F-127, 7.9 +/- 0.7 h. We conclude that XRF is an easy, reliable, noninvasive method to monitor the retention of antigens in these adjuvant solutions. PMID- 11416999 TI - Effects of formulation variables and post-compression curing on drug release from a new sustained-release matrix material: polyvinylacetate-povidone. AB - A new commercially available sustained-release matrix material, Kollidon SR, composed of polyvinylacetate and povidone, was evaluated with respect to its ability to modulate the in vitro release of a highly water-soluble model compound, diphenhydramine HCl. Kollidon SR was found to provide a sustained release effect for the model compound, with certain formulation and processing variables playing an important role in controlling its release kinetics. Formulation variables affecting the release include the level of the polymeric material in the matrix, excipient level, as well as the nature of the excipients (water soluble vs. water insoluble). Increasing the ratio of a water-insoluble excipient, Emcompress, to Kollidon SR enhanced drug release. The incorporation of a water-soluble excipient, lactose, accelerated its release rate in a more pronounced manner. Stability studies conducted at 40 degrees C/75% RH revealed a slow-down in dissolution rate for the drug-Kollidon SR formulation, as a result of polyvinylacetate relaxation. Further studies demonstrated that a post compression curing step effectively stabilized the release pattern of formulations containing > or = 47% Kollidon SR. The release mechanism of Kollidon drug and drug-Kollidon-Emcompress formulations appears to be diffusion controlled, while that of the drug-Kollidon-lactose formulation appears to be controlled predominantly by diffusion along with erosion. PMID- 11417000 TI - Characterization of a backbone cleavage product of BMS-196854 (Oncostatin M), a recombinant anti-inflammatory cytokine. AB - PURPOSE: BMS-196843 (Oncostatin M) is a therapeutic recombinant protein in development. Scale-up process changes led to unexpected instability of the bulk drug substance solution during storage. A product with an apparent higher MW than the parent protein was observed by the size-exclusion chromatography (SEC). This study was aimed to fully characterize the product and to identify a solution to stabilize the protein. METHODS: SEC, SDS-PAGE, tryptic mapping, and N-terminal sequencing were performed to characterize the unknown product. The effect of pH, temperature, bulk concentration, and immobilized trypsin inhibitor on the degradation rate was studied to elucidate the mechanism and to identify stabilization strategies. RESULTS: Despite the apparent high MW indicated initially by SEC, the unknown was characterized to be a degradation product resulted from a backbone cleavage between residues Arg145-Gly146. The resulting fragments from the backbone cleavage were, however, still linked through an intramolecular disulfide bond. Thus, the final product had a more open structure with an increased hydrodynamic radius compared to the parent protein, which explains the initial SEC results. The site-specific backbone cleavage was suspected to be catalyzed by trypsin-like protease impurities in the bulk solution. The bulk drug substance solution was subsequently treated with immobilized soybean trypsin inhibitor, and the degradation rate was significantly reduced. Furthermore, increasing the solution pH from 5 to 8 led to an increase in the degradation rate, which was consistent with the expected pH dependency of trypsin activity. In addition, the effect of bulk concentration also supported the involvement of protease impurities rather than a spontaneous peptide bond hydrolysis reaction. CONCLUSION: Trace trypsin-like protease impurities led to an unusual site-specific backbone cleavage of BMS-196854. The proteolytic degradation can be minimized by treating the bulk solution with immobilized soybean trypsin inhibitor and/or controlling the solution pH and storage temperature. PMID- 11417001 TI - Properties of tablets containing granulations of ibuprofen and an acrylic copolymer prepared by thermal processes. AB - The objective of this study was to investigate the properties of tablets containing granulations of ibuprofen (Ibu) and Ammonio Methacrylate Copolymer, Type B (Eudragit RS PO) prepared by hot-melt processing. Tablets were compressed from granules prepared by hot-melt granulation (HMG) or direct compression (DC). For the hot-melt extrusion (HME) process, tablets were prepared by cutting the extrudate, manually. The physicochemical properties of tablets were investigated using thermal analysis, powder X-ray diffraction analysis, tablet hardness, and drug dissolution. The effect of thermal treatment of tablets on the dissolution characteristics of Ibu was also investigated. The results demonstrated that the Ibu lowered the glass transition temperature (Tg) of the Eudragit RS PO and the softened polymer functioned as a thermal binder in the granulation. Ibu was demonstrated to be an effective plasticizer for Eudragit RS PO in the thermal processes. The efficiency of the granulation process increased with increasing levels of Eudragit RS PO in the powder blend. Higher levels of Eudragit RS PO in the tablets prepared by HMG or HME resulted in a decrease in the dissolution rate of the Ibu. An increase in the amount of Ibu in the tablets prepared by HMG or DC led to a decrease in the initial dissolution rate of the Ibu. Following the thermal treatment of the Ibu tablets prepared by HMG, the dissolution rate was significantly decreased due to structural changes in the tablets that resulted from the fusion and coalescence of plasticized polymer particles, causing a reduction in tablet porosity. The Ibu tablets prepared by HME demonstrated minimal changes in their release properties following thermal treatment even at temperatures higher than the Tg of the polymer. HME was shown to be a novel method to prepare matrix tablets and stable dissolution properties were obtained when tablets were stored at 40 degrees C for 30 days. PMID- 11417002 TI - The effects of plasticizers and titanium dioxide on the properties of poly(vinyl alcohol) coatings. AB - Poly(vinyl alcohol) has not previously been examined in much detail as a controlled release polymer for use in pharmaceutical formulations. However, this food grade polymer has barrier and tensile properties which make it attractive for such applications. The effects of several diluents and fillers on Poly(vinyl alcohol) (PVAL) coatings have been determined using both mechanical property and water vapor permeability measurements. It has been found that the alcohol ethoxylate Neodol 23-6.5 (CH3(CH2)11-O-(CH2-CH2-O)6-H) acts as a plasticizer for PVAL only up to 15-20 wt% in contrast to 600 molecular weight Polyethylene Glycol (PEG 600), which continuously plasticizes PVAL. The effects of Neodol on PVAL mechanical properties and water vapor permeability at higher concentrations can be explained in terms of Neodol phase separation and has been confirmed with DSC. The inert filler and whitener titanium dioxide (TiO2) monotonically degrades film mechanical properties and increases water vapor permeability of the coating. Attempts to correlate coating dust generated during particle attrition tests with mechanical property measurements were unsuccessful. A correlation between accelerated granule stability and water vapor permeability of the PVAL coating was established. PMID- 11417003 TI - Drugs of the future: lessons of the past. PMID- 11417004 TI - Ethics: a unifying force. The Tavistock Principles provide a decisionmaking framework for everyone. AB - A multidisciplinary group of health care leaders has developed a set of seven ethical principles that can provide everyone in an organization with a common language. Trustees should find the principles particularly helpful for setting policy. PMID- 11417005 TI - Filling the information gap. Not-for-profits are gradually opening up to bondholders. AB - Once the exclusive domain of the corporate world, now not-for-profit health care organizations are gravitating toward greater financial disclosure. The reason: a growing consensus among boards and executives that accountability boosts investor confidence and may pay off in better bond ratings. PMID- 11417006 TI - Grassroots advocacy is your responsibility. PMID- 11417007 TI - Quality. Board leadership for patient safety: new JCAHO standards. PMID- 11417008 TI - Workforce supply for hospitals and health systems. PMID- 11417011 TI - Nurses needed--stat! The hospital field is desperate for creative solutions to stop the nursing drain. AB - Reaching epidemic proportions, the nurse shortage may be the worst and most far reaching problem hospitals have confronted in the last 10 years--a decade beset by health care woes. Hopefully, once trustees understand the reasons for the shortage, they can devise ways to alleviate it. PMID- 11417012 TI - Training police officers to help people with mental illnesses. PMID- 11417013 TI - Before the experts arrive. AB - This article addresses the rare event of the hostage situation in a forensic psychiatric nursing setting. It has a specific focus on the initial response and the accompanying issues for nurses at the clinical interface as the situation emerges. The intention of this article is to both offer guidance and raise the profile of this unique management issue because little attention is drawn to early-stage hostage situations at an organizational level. When a hostage situation occurs, inadequate preparation can mean the difference between life and death for the hostage, negotiator, or hostage taker. This article provides an overview of the relevant literature and offers guidance about the actions required when a nurse suddenly becomes responsible for managing the early stages of such a traumatic event. Responses, safety, and communication factors concerning the hostage taker are covered. PMID- 11417014 TI - Therapeutic interactions in a medication education group. Using the psychopharmacology RACE. AB - 1. The Psychopharmacology RACE is an interactive, innovative game that emphasizes group teaching about psychiatric medications and disorders at the clients' levels of functioning. 2. Implementation of the Psychopharmacology RACE provides the opportunity to structure a medication group, impart information, and facilitate the development of therapeutic group factors. 3. Interactive group learning with the Psychopharmacology RACE can be a useful tool to enhance learning when clients are unable to read and understand written materials. PMID- 11417015 TI - The cognitive model. Interventions for improved patient-provider communication. AB - 1. Improving patient-provider interactions mandates an understanding on the part of health care providers of patients' situations, thoughts, feelings, and behaviors. 2. The cognitive model, a cornerstone of cognitive therapy, provides a framework to understand the patient's perspective of his or her situation or illness, which is a necessary component of effective patient-provider interactions. 3. Distance learning formats provide flexibility in meeting continuing education needs and allow learners and presenters to interact in a virtual mode. PMID- 11417016 TI - Sexual misconduct by health care providers. AB - 1. Just as in society in general, health care providers can engage in compulsive sexual behaviors that jeopardize their health, careers, and relationships, both personal and professional. 2. Sex addicts not only deny and rationalize their acts but also resist accepting any feedback about their sexually inappropriate behaviors. 3. To protect patients from sexual misconduct, health care providers who exhibit sexually inappropriate behaviors must be confronted, and consistent limits must be set, including terminating patient contact, if necessary. 4. Treatment for sexually compulsive behaviors is multifaceted, and signs that a person is cooperating with treatment are ceasing the compulsive behaviors, experiencing feelings of remorse, expressing regret, accepting personal responsibility, and abiding by guidelines for reinstatement. PMID- 11417017 TI - A new day. PMID- 11417019 TI - Harm, ethics committees and the gene therapy death. PMID- 11417020 TI - Sham neurosurgery in patients with Parkinson's disease: is it morally acceptable? AB - For a few decades, patients with Parkinson's disease (PD) have been treated with intracerebral transplantations of fetal mesencephalic tissue. The results of open trials have been variable. Double blind, placebo-controlled studies have recently been started in order to further investigate the efficacy of this new medical technique. In this paper we challenge the need for sham surgery in neurotransplantation research on PD patients. Considerations regarding the research subjects' informed consent, therapeutic misconception, the integrity of the human body, and the assessment of risks and benefits argue against sham surgery for patients with PD. Moreover, there is an alternative, less harmful mode of research that can provide the same or comparable scientific evidence. A plea is made for intrapatient research based on quantitative measurements of the patient's pre- and post-operative condition combined with similar research on a reference group of patients who have received the standard treatment. PMID- 11417021 TI - The process of informed consent for urgent abdominal surgery. AB - OBJECTIVES: To assess perceptions of the informed consent process in patients undergoing urgent abdominal surgery. DESIGN: A prospective observational study was carried out using structured questionnaire-based interviews. Patients who had undergone urgent abdominal surgery were interviewed in the postoperative period to ascertain their perceptions of the informed consent process. Replies were compared to responses obtained from a control group undergoing elective surgery, to identify factors common to the surgical process and those specific to urgent surgery. Patients' perceptions of received information were also compared to the information perceived to have been provided by the consent obtainers. SETTING: Gastrointestinal surgical service of a university teaching hospital. PATIENTS: Seventy-four consecutive patients undergoing urgent abdominal surgery and 80 control patients undergoing elective surgery. MAIN MEASUREMENTS: Principal outcome measures were patients perceptions of factors interfering with the ability to give informed consent, assessment of the quality of informed consent and the degree of discussion of the expected outcomes. RESULTS: Forty-nine of the seventy-four (66%) patients undergoing urgent surgery perceived that pain did not affect their ability to give informed consent. Twenty-seven reported an adverse effect of analgesia on the ability to give informed consent. Only 22% of patients undergoing urgent surgery perceived that there had been any discussion of potential side effects and complications of surgery. CONCLUSION: The majority of patients in this series with acute intra-abdominal surgical conditions perceive that they retain the ability to give informed consent for surgery. There is a need for improved discussion of therapeutic options and likely outcomes. PMID- 11417022 TI - It's only love? Some pitfalls in emotionally related organ donation. AB - Transplanting organs from emotionally related donors has become a fairly routine procedure in many countries. However, donors have to be chosen carefully in order to avoid not just medically, but also morally, questionable outcomes. This paper draws attention to vulnerabilities that may affect the voluntariness of the donor's decision. Suggestions are made as to how to approach the evaluation and selection of potential donors. PMID- 11417023 TI - Is current practice around late termination of pregnancy eugenic and discriminatory? Maternal interests and abortion. AB - The attitudes of Australian practitioners working in clinical genetics and obstetrical ultrasound were surveyed on whether termination of pregnancy (TOP) should be available for conditions ranging from mild to severe fetal abnormality and for non-medical reasons. These were compared for terminations at 13 weeks and 24 weeks. It was found that some practitioners would not facilitate TOP at 24 weeks even for lethal or major abnormalities, fewer practitioners support TOP at 24 weeks compared with 13 weeks for any condition, and the difference in attitudes to TOP between 13 weeks and 24 weeks is most marked for pregnancies which are normal or involve a mild disorder. It is argued that a fetal abnormality criterion for late TOP is inconsistently applied, discriminatory and eugenic. Four possible moral justifications for current practice are examined, each of which would require significant changes to current practice. I argue in favour of a maternal interests criterion for any TOP. PMID- 11417024 TI - Methodological quality and reporting of ethical requirements in clinical trials. AB - OBJECTIVES: To assess the relationship between the approval of trials by a research ethics committee (REC) and the fact that informed consent from participants (ICP) was obtained, with the quality of study design and methods. DESIGN: Systematic review using a standardised checklist. MAIN MEASURES: Methodological and ethical issues of all trials published between 1993 and 1995 in the New England Journal of Medicine, the Lancet, the Journal of the American Medical Association and the British Medical Journal were studied. In addition, clinical trials conducted in Spain and published by at least one Spanish author during the same period in any other journal were also included. RESULTS: We studied the published articles of 767 trials and found the following indicators of lower methodological quality to be independent predictors for failure to disclose REC approval or ICP: absence of concealment of allocation, lack of justification for unblinded trials, not using a treatment for the patients in the control group, absent information on statistical methods, not including sample size estimation, not establishing the rules to stop the trial, and omitting the presentation of a baseline comparison of groups. CONCLUSION: Trials of higher methodological and scientific quality were more likely to provide information about their ethical aspects. PMID- 11417025 TI - Randomised clinical trials: a source of ethical dilemmas. AB - Advances in medicine are closely linked to clinical research, but certain study procedures may be in conflict with the fundamental principles of ethics and codes of conduct in medicine. Following an analysis of two studies involving treatments for acute myocardial infarction (AMI), the admissibility of continuing a study was questioned after the initial results for two types of treatment showed that one was significantly better than the other. Also considered doubtful was the information provided to patients with the object of obtaining their informed consent. PMID- 11417026 TI - Rationing and life-saving treatments: should identifiable patients have higher priority? AB - Health care systems across the world are unable to afford the best treatment for all patients in all situations. Choices have to be made. One key ethical issue that arises for health authorities is whether the principle of the "rule of rescue" should be adopted or rejected. According to this principle more funding should be available in order to save lives of identifiable, compared with unidentifiable, individuals. Six reasons for giving such priority to identifiable individuals are considered. All are rejected. It is concluded that the principle of the rule of rescue should not be used in determining the allocation of health resources. PMID- 11417027 TI - Doctors' authoritarianism in end-of-life treatment decisions. A comparison between Russia, Sweden and Germany. AB - OBJECTIVES: The study was performed in order to investigate how end-of-life decisions are influenced by cultural and sociopolitical circumstances and to explore the compliance of doctors with patient wishes. PARTICIPANTS AND MEASUREMENT: Five hundred and thirty-five physicians were surveyed in Sweden (Umea), Germany (Rostock and Neubrandenburg), and in Russia (Arkhangelsk) by a questionnaire. The participants were recruited according to availability and are not representative. The questionnaire is based on the one developed by Molloy and co-workers in Canada which contains three case vignettes about an 82-year-old Alzheimer patient with an acute life-threatening condition; the questionnaire includes different levels of information about his treatment wishes. We have added various questions about attitudes determining doctors' decision making process (legal and ethical concerns, patient's and family wishes, hospital costs, patient's age and level of dementia and physician's religion). RESULTS: Swedish physicians chose fewer life-prolonging interventions as compared with the Russian and the German doctors. Swedish physicians would perform cardiopulmonary resuscitation (CPR) in the event of a cardiac arrest less frequently, followed by the German doctors. More than half the Russian physicians decided to perform CPR irrespective of the available information about the patient's wishes. Level of dementia emerged as the most powerful determining attitude-variable for the decision making in all three countries. CONCLUSIONS: The lack of compliance with patient wishes among a substantial number of doctors points to the necessity of emphasising ethical aspects both in medical education and clinical practice. The inconsistency in the treatment decisions of doctors from different countries calls for social consensus in this matter. PMID- 11417028 TI - Truth-telling and patient diagnoses. AB - How do physicians handle informing patients of their diagnoses and how much information do patients really want? How do registered nurses view both sides of this question? Three questionnaires were constructed and administered in a mid size hospital in New York state. Physicians and nurses underestimate the number of patients who want detailed information. Patients who earn more than average, have a college education, and who are under age 60 are more likely to want information, and state that their physician should give it to them. Only 42% of physicians state that patients want a detailed description of their diagnosis and treatment options. Physicians educated outside the USA appeared to be more likely to change their criteria for informing patients and, along with American-educated nurses, were more willing to participate in formal discussions of the issue. Physicians should comply with the wishes of patients for information and include them in the team deciding on diagnosis and treatment. PMID- 11417029 TI - That's another story: narrative methods and ethical practice. AB - This paper examines the use of case studies in ethics education. While not dismissing their value for specific purposes, the paper shows the limits of their use. While agreeing that case studies are narratives, although rather thin stories, the paper argues that the claim that case studies could represent reality is difficult to sustain. Instead, the paper suggests a way of using stories in ethics teaching that could be more real for students, while also giving them a way of thinking about their own professional practices. The paper shows how the method can be used to develop a more critical and reflective practice for students in the health care professions. Some immediate problems with the method are discussed. PMID- 11417030 TI - Misoprostol in a topsyturvy world. PMID- 11417031 TI - Health and intellectual property rights. PMID- 11417032 TI - Attitudes towards war, killing, and punishment of children among young people in Estonia, Finland, Romania, the Russian Federation, and the USA. AB - OBJECTIVE: To study the cultural differences in moral disengagement, which lends support to attitudes used to justify violence. METHODS: We carried out classroom surveys of a total of 3122 students in the USA (Houston, TX, and Washington, DC) and in four European countries--Estonia (Tartu), Finland (Helsinki), Romania (Satu Mare) and the Russian Federation (St Petersburg). Data were also taken from a random sample telephone survey of 341 young adults (aged 18-35 years) in Texas, USA. Ten distinct groups were studied. Seven questions were common to all the surveys, using identical statements about the participants' agreement with attitudes relating to war, diplomacy, killing, and the punishment of children. FINDINGS: The US students were more likely than those in Europe to agree with the following statements: "War is necessary" (20% vs 9%), "A person has the right to kill to defend property" (54% vs 17%), and "Physical punishment is necessary for children" (27% vs 10%). Justification of war and killing was less common among females than males in all groups; other differences within the US groups and the European groups were smaller than the differences between the US and European groups. CONCLUSION: The results confirm the gap between the US and European groups in moral disengagement attitudes and tendencies that could lead to deadly violence. PMID- 11417033 TI - Comparison of generic and proprietary sodium stibogluconate for the treatment of visceral leishmaniasis in Kenya. AB - OBJECTIVE: To compare the use of generic and proprietary sodium stibogluconate for the treatment of visceral leishmaniasis (kala-azar). METHODS: A total of 102 patients with confirmed kala-azar were treated in a mission hospital in West Pokot region, Kenya, with sodium stibogluconate (20 mg/kg/day for 30 days)- either as Pentostam (PSM) or generic sodium stibogluconate (SSG); 51 patients were allocated alternately to each treatment group. FINDINGS: There were no significant differences in baseline demographic characteristics or disease severity, or in events during treatment. There were 3 deaths in the PSM group and 1 in the SSG group; 2 patients defaulted in each group. Only 1 out of 80 test-of cure splenic aspirates was positive for Leishmania spp.; this patient was in the SSG group. Follow-up after > or = 6 months showed that 6 out of 58 patients had relapsed, 5 in the SSG group and 1 in the PSM group. No outcome variable was significantly different between the two groups. CONCLUSION: The availability of cheaper generic sodium stibogluconate, subject to rigid quality controls, now makes it possible for the health authorities in kala-azar endemic areas to provide treatment to many more patients in Africa. PMID- 11417034 TI - Impact of prepackaging antimalarial drugs on cost to patients and compliance with treatment. AB - OBJECTIVE: To examine the extent to which district health teams could reduce the burden of malaria, a continuing major cause of mortality and morbidity, in a situation where severe resource constraints existed and integrated care was provided. METHODS: Antimalarial drugs were prepackaged into unit doses in an attempt to improve compliance with full courses of chemotherapy. FINDINGS: Compliance improved by approximately 20% in both adults and children. There were 50% reductions in cost to patients, waiting time at dispensaries and drug wastage at facilities. The intervention, which tended to improve both case and drug management at facilities, was well accepted by health staff and did not involve them in additional working time. CONCLUSION: The prepackaging of antimalarials at the district level offers the prospect of improved compliance and a reduction in the spread of resistance. PMID- 11417035 TI - Estimates of the maternal mortality ratio in two districts of the Brong-Ahafo region, Ghana. AB - OBJECTIVE: To estimate the maternal mortality ratio (MMR) by the sisterhood method in two districts of the Brong-Ahafo region of Ghana, and to determine the impact of different assumptions and analytical decisions on these estimates. METHODS: Indirect estimates of the MMR were calculated from data collected in 1995 by Family Health International (FHI) on 5202 women aged 15-49 years, using a household screen of randomly selected areas in the two districts. Other data from the nationally representative 1994 Ghana Infant, Child and Maternal Mortality Survey (ICMMS) and from the 1997 Kassena-Nankana District study were also used for comparison. FINDINGS: Based on the FHI data, the MMR was estimated to be 269 maternal deaths per 100,000 live births for both districts combined, a figure higher than ICMMS estimates. Biases during data collection may account for this difference, including the fact that biases underestimating mortality are more common than those overestimating it. Biases introduced during data analysis were also considered, but only the total fertility rate used to calculate the MMR seemed to affect the estimates significantly. CONCLUSIONS: The results indicate that the sisterhood method is still being refined and the extent and impact of biases have only recently received attention. Users of this method should be aware of limitations when interpreting results. We recommend using confidence limits around estimates, both to dispel false impressions of precision and to reduce overinterpretation of data. PMID- 11417036 TI - Nested case-control study in a serological survey to evaluate the effectiveness of a Chagas disease control programme in Brazil. AB - OBJECTIVE: To identify risk factors associated with Trypanosoma cruzi infections in areas under surveillance in the State of Minas Gerais, Brazil. METHODS: A model using a nested case-control design incorporated within a serological survey of schoolchildren which was employed to evaluate the effectiveness of the Chagas disease control programme. FINDINGS: In a sample of 40,374 schoolchildren (aged 7 14 years) surveyed, 16 children tested positive for T. cruzi antibody (by indirect immunofluorescence and indirect haemagglutination). In the case-control study, each case was randomly matched to three seronegative controls (classroom and age +/- 1 year). Compared to controls, T. cruzi-seropositive children were more likely to have a seropositive mother (odds ratio (OR) = 6.8; 95% confidence interval (CI) = 0.71-63.9) or a seropositive family member (OR = 8.6; 95% CI = 1.0-75.5). CONCLUSION: Use of the nested case-control model in a sero epidemiological survey to evaluate risk factors for T. cruzi transmission was adequate for assessing the effectiveness of a Chagas disease control programme. PMID- 11417037 TI - Prevalence of vitamin A deficiency in children aged 6-9 years in Wukro, northern Ethiopia. AB - OBJECTIVE: To determine the prevalence of vitamin A deficiency in children aged 6 9 years in northern Ethiopia. METHODS: A cross-sectional study was carried out and the data were analysed for 824 (61.5%) of 1339 eligible children for whom there was complete information on biochemical vitamin A status, dietary vitamin A intake, ocular examination for xerophthalmia, and anthropometry. FINDINGS: The prevalence of xerophthalmia was 5.8%; serum retinol levels were below 0.35 mumol/l and between 0.35 and 0.70 mumol/l in 8.4% and 51.1% of the children respectively. The liver vitamin A reserve (modified relative dose response ratio > or = 0.06) was low in 41.0% of the children. CONCLUSION: The high prevalence of severe vitamin A deficiency in children aged 6-9 years indicates the need to reevaluate the practice of targeting vitamin A supplementation programmes on children under 6 years of age in areas where vitamin A deficiency is endemic. PMID- 11417038 TI - Critical control points of complementary food preparation and handling in eastern Nigeria. AB - OBJECTIVE: To investigate microbial contamination and critical control points (CCPs) in the preparation and handling of complementary foods in 120 households in Imo state, Nigeria. METHODS: The Hazard Analysis Critical Control Point (HACCP) approach was used to investigate processes and procedures that contributed to microbial contamination, growth and survival, and to identify points where controls could be applied to prevent or eliminate these microbiological hazards or reduce them to acceptable levels. Food samples were collected and tested microbiologically at different stages of preparation and handling. FINDINGS: During cooking, all foods attained temperatures capable of destroying vegetative forms of food-borne pathogens. However, the risk of contamination increased by storage of food at ambient temperature, by using insufficiently high temperatures to reheat the food, and by adding contaminated ingredients such as dried ground crayfish and soybean powder at stages where no further heat treatment was applied. The purchasing of contaminated raw foodstuffs and ingredients, particularly raw akamu, from vendors in open markets is also a CCP. CONCLUSION: Although an unsafe environment poses many hazards for children's food, the hygienic quality of prepared food can be assured if basic food safety principles are observed. When many factors contribute to food contamination, identification of CCPs becomes particularly important and can facilitate appropriate targeting of resources and prevention efforts. PMID- 11417039 TI - The impact of a clinical training unit on integrated child health care in Mexico. AB - This study had two aims: to describe the activities of a clinical training unit set up for the integrated management of sick children, and to evaluate the impact of the unit after its first four years of operation. The training unit was set up in the outpatient ward of a government hospital and was staffed by a paediatrician, a family medicine physician, two nurses and a nutritionist. The staff kept a computerized database for all patients seen and they were supervised once a month. During the first three years, the demand for first-time medical consultation increased by 477% for acute respiratory infections (ARI) and 134% for acute diarrhoea (AD), with an average annual increase of demand for medical care of 125%. Eighty-nine per cent of mothers who took their child for consultation and 85% of mothers who lived in the catchment area and had a deceased child received training on how to recognize alarming signs in a sick child. Fifty-eight per cent of these mothers were evaluated as being properly trained. Eighty-five per cent of primary care physicians who worked for government institutions (n = 350) and 45% of private physicians (n = 90) were also trained in the recognition and proper management of AD and ARI. ARI mortality in children under 1 year of age in the catchment area (which included about 25,000 children under 5 years of age) decreased by 43.2% in three years, while mortality in children under 5 years of age decreased by 38.8%. The corresponding figures for AD mortality reduction were 36.3% and 33.6%. In this same period, 11 clinical research protocols were written. In summary, we learned that a clinical training unit for integrated child care management was an excellent way to offer in-service training for primary health care physicians. PMID- 11417040 TI - Focusing strategies of condom use against HIV in different behavioural settings: an evaluation based on a simulation model. AB - Using a sexually transmitted diseases simulation model (STDSIM), we made projections of HIV spread for four profiles of sexual behaviour reflecting patterns encountered across the developing world: 1) much commercial sex, no short relationships; 2) commercial sex, concurrent short relationships; 3) concurrent relationships, no commercial sex; 4) serial short relationships, some commercial sex. We studied the effects of increasing condom use in three target groups: commercial sex workers (CSWs); men engaging in commercial contacts and short relationships; and females in steady relationships. The projections indicated that the CSW and male strategies were more effective in reducing HIV incidence than the strategy focusing on females in steady relationships. In the long run, even the group of men and women with one recent partner were better protected against HIV infection by condom use in high-risk contacts than by condom use in steady relationships. Furthermore, the numbers of HIV cases prevented per condom used were 7 to 500 times higher for condoms used by CSWs or men engaging in short relationships and commercial sex than for ones used by females in steady relationships. The results indicated the merit of focusing on high-risk groups irrespective of the pattern of sexual behaviour, even in epidemics that had already spread throughout populations. PMID- 11417041 TI - Transmission of Helicobacter pylori: a role for food? AB - Helicobacter pylori colonizes and grows in human gastric epithelial tissue and mucus. Its presence is associated with gastritis and there is substantial evidence that it causes peptic and duodenal ulcers and chronic gastritis. Since 1994, H. pylori has been classified as carcinogenic to humans. In industrialized countries, as many as 50% of adults are infected with the pathogen, while in the developing world, prevalence values of about 90% have been reported. As little is known about the mode of transmission, a literature search was carried out to determine whether food acts a reservoir or vehicle in the transmission of H. pylori. Although growth of the pathogen should be possible in the gastrointestinal tract of all warm-blooded animals, the human stomach is its only known reservoir. Under conditions where growth is not possible, H. pylori can enter a viable, but nonculturable state. H. pylori has been detected in such states in water, but not in food. Person-to-person contact is thought to be the most likely mode of transmission, and there is no direct evidence that food is involved in the transmission of H. pylori. PMID- 11417042 TI - Impact of the World Trade Organization TRIPS agreement on the pharmaceutical industry in Thailand. AB - The 1994 World Trade Organization (WTO) Agreement on Trade-Related Aspects of Intellectual Property Rights (TRIPS) established minimum universal standards in all areas of intellectual property. It is intended to implement these standards globally through a WTO enforcement mechanism. The present article proposes a strategy for alleviating the potentially negative impact of TRIPS in Thailand in relation to the following: purchasers; prescribers and dispensers; producers; products; price control; patent-to-third-party; parallel imports; power of the customer; patentable new drugs; personnel; and prevention policies. The following TRIPS provisions are pertinent to the pharmaceutical industry in Thailand: the limited term of product and process patents; the conditions of protection; and the broad scope for compulsory licensing and enforcement procedures in the national patent system. PMID- 11417043 TI - A revolution in the management of diarrhoea. PMID- 11417044 TI - The challenges facing Third World countries in banning tobacco. PMID- 11417045 TI - Interpretation of the Mwanza and Rakai STI trials. PMID- 11417046 TI - Interpretation of the Mwanza and Rakai STD trials. PMID- 11417047 TI - The world in 2050: more crowded, urban and aged. PMID- 11417048 TI - AIDS vaccine research riding high. PMID- 11417049 TI - Water and health, hand-in-hand for a day. PMID- 11417050 TI - A day in the life of the world's anti-TB drive. PMID- 11417051 TI - Wealth for health--a Gates perspective. Interview by Barry Whyte. PMID- 11417052 TI - A computational model of action selection in the basal ganglia. I. A new functional anatomy. AB - We present a biologically plausible model of processing intrinsic to the basal ganglia based on the computational premise that action selection is a primary role of these central brain structures. By encoding the propensity for selecting a given action in a scalar value (the salience), it is shown that action selection may be recast in terms of signal selection. The generic properties of signal selection are defined and neural networks for this type of computation examined. A comparison between these networks and basal ganglia anatomy leads to a novel functional decomposition of the basal ganglia architecture into 'selection' and 'control' pathways. The former pathway performs the selection per se via a feedforward off-centre on-surround network. The control pathway regulates the action of the selection pathway to ensure its effective operation, and synergistically complements its dopaminergic modulation. The model contrasts with the prevailing functional segregation of basal ganglia into 'direct' and 'indirect' pathways. PMID- 11417053 TI - A computational model of action selection in the basal ganglia. II. Analysis and simulation of behaviour. AB - In a companion paper a new functional architecture was proposed for the basal ganglia based on the premise that these brain structures play a central role in behavioural action selection. The current paper quantitatively describes the properties of the model using analysis and simulation. The decomposition of the basal ganglia into selection and control pathways is supported in several ways. First, several elegant features are exposed--capacity scaling, enhanced selectivity and synergistic dopamine modulation--which might be expected to exist in a well designed action selection mechanism. The discovery of these features also lends support to the computational premise of selection that underpins our model. Second, good matches between model globus pallidus external segment output and globus pallidus internal segment and substantia nigra reticulata area output, and neurophysiological data, have been found which are indicative of common architectural features in the model and biological basal ganglia. Third, the behaviour of the model as a signal selection mechanism has parallels with some kinds of action selection observed in animals under various levels of dopaminergic modulation. PMID- 11417054 TI - Biomechanical analysis of movement strategies in human forward trunk bending. I. Modeling. AB - Two behavioral goals are achieved simultaneously during forward trunk bending in humans: the bending movement per se and equilibrium maintenance. The objective of the present study was to understand how the two goals are achieved by using a biomechanical model of this task. Since keeping the center of pressure inside the support area is a crucial condition for equilibrium maintenance during the movement, we decided to model an extreme case, called "optimal bending", in which the movement is performed without any center of pressure displacement at all, as if standing on an extremely narrow support. The "optimal bending" is used as a reference in the analysis of experimental data in a companion paper. The study is based on a three-joint (ankle, knee, and hip) model of the human body and is performed in terms of "eigenmovements", i.e., the movements along eigenvectors of the motion equation. They are termed "ankle", "hip", and "knee" eigenmovements according to the dominant joint that provides the largest contribution to the corresponding eigenmovement. The advantage of the eigenmovement approach is the presentation of the coupled system of dynamic equations in the form of three independent motion equations. Each of these equations is equivalent to the motion equation for an inverted pendulum. Optimal bending is constructed as a superposition of two (hip and ankle) eigenmovements. The hip eigenmovement contributes the most to the movement kinematics, whereas the contributions of both eigenmovements into the movement dynamics are comparable. The ankle eigenmovement moves the center of gravity forward and compensates for the backward center of gravity shift that is provoked by trunk bending as a result of dynamic interactions between body segments. An important characteristic of the optimal bending is the timing of the onset of each eigenmovement: the ankle eigenmovement onset precedes that of the hip eigenmovement. Without an earlier onset of the ankle eigenmovement, forward bending on the extremely narrow support results in falling backward. This modeling approach suggests that during trunk bending, two motion units--the hip and ankle eigenmovements--are responsible for the movement and for equilibrium maintenance, respectively. PMID- 11417055 TI - Biomechanical analysis of movement strategies in human forward trunk bending. II. Experimental study. AB - The large mass of the human upper trunk, its elevated position during erect stance, and the small area limited by the size of the feet, stress the importance of equilibrium control during trunk movements. The objective of the present study was to perform a biomechanical analysis of fast forward trunk movements in order to understand the coordination between movement and posture. The analysis is based on a comparison between experimentally observed bending and hypothetical "optimal bending" performed on an infinitely narrow support, as presented in a companion paper. The experimental data were obtained from 16 subjects who performed fast forward bending while standing on a wide platform or on a narrow beam. The analysis is performed by decomposition of the movement into three dynamically independent components, each representing a movement along one of the three eigenvectors of the motion equation. The eigenmovements are termed "hip", "ankle", and "knee" eigenmovements, according to the dominant joint. The experimentally observed movement is characterized mainly by the hip and ankle eigenmovements, whereas the knee eigenmovement is negligible. Similarly to the "optimal bending" the ankle eigenmovement starts earlier and lasts longer than the hip eigenmovement. An early forward acceleration of the center of gravity in the ankle eigenmovement is caused by anticipatory changes in the ankle joint torque. This clarifies the role of the early tibialis anterior burst and/or soleus inhibition usually observed in electromyographic recordings during forward bending. The results suggest that the hip and the ankle eigenmovements can be treated as independently controlled motion units aimed at functionally different behavioral goals: the bending per se and postural adjustment. It is proposed that the central nervous system has to control these motion units sequentially in order to perform the movement and maintain equilibrium. It is also suggested that the hip and ankle eigenmovements can be regarded as a biomechanical background for the hip and ankle strategies introduced by Horak and Nashner (1986) on the basis of electromyographic recordings and kinematic patterns in response to postural perturbations. PMID- 11417056 TI - Simulation of plasticity in the adult visual cortex. AB - Retinal plasticity has been shown in the adult visual nervous system in mammals. Following a retinal lesion (scotoma) there is a reorganization of the cortical receptive field distribution: cortical neurons selective to visual stimuli in the area of the visual field corresponding to the retinal lesion, become selective to other parts of the visual field. In this work, we study this effect with a self organizing neural network. In a first stage, the network reaches a pattern of connectivity that represents normal development of neuronal selectivity. The scotoma is simulated by perturbing accordingly the properties of a region of the input layer representing the retina. The system evolves to a new receptive field distribution mainly by means of the reorganization of the intra cortical connectivity. No major change of the geniculo cortical connectivity is detected. This may explain the surprisingly short time scale of the event. PMID- 11417057 TI - Interactions between eye and head control signals can account for movement kinematics. AB - Advances in understanding the neural control of saccades (visual orienting movements made when the head is prevented from moving) stem largely from early modeling efforts which provided a framework for developing and testing hypotheses about the relationships between neural activity and observed behaviors. When the head is free to move, visual orienting is often accomplished with coordinated movements of the eyes and head. A recent description of the temporal progression (i.e., kinematics) of these movements led to the hypothesis that eye and head control signals interact. This hypothesis is now formalized as a control systems model which accounts for existing data and makes explicit predictions about the neural control of orienting gaze shifts. PMID- 11417058 TI - Partial directed coherence: a new concept in neural structure determination. AB - This paper introduces a new frequency-domain approach to describe the relationships (direction of information flow) between multivariate time series based on the decomposition of multivariate partial coherences computed from multivariate autoregressive models. We discuss its application and compare its performance to other approaches to the problem of determining neural structure relations from the simultaneous measurement of neural electrophysiological signals. The new concept is shown to reflect a frequency-domain representation of the concept of Granger causality. PMID- 11417059 TI - Enhancing human balance control with galvanic vestibular stimulation. AB - With galvanic vestibular stimulation (GVS), electrical current is delivered transcutaneously to the vestibular afferents through electrodes placed over the mastoid bones. This serves to modulate the continuous firing levels of the vestibular afferents, and causes a standing subject to lean in different directions depending on the polarity of the current. Our objective in this study was to test the hypothesis that the sway response elicited by GVS can be used to reduce the postural sway resulting from a mechanical perturbation. Nine subjects were tested for their postural responses to both galvanic stimuli and support surface translations. Transfer-function models were fit to these responses and used to calculate a galvanic stimulus that would act to counteract sway induced by a support-surface translation. The subjects' responses to support-surface translations, without and with the stabilizing galvanic stimulus, were then measured. With the stabilizing galvanic stimulus, all subjects showed significant reductions in both sway amplitude and sway latency. Thus, with GVS, subjects maintained a more erect stance and followed the support-surface displacement more closely. These findings suggest that GVS could possibly form the basis for a vestibular prosthesis by providing a means through which an individual's posture can be systematically controlled. PMID- 11417060 TI - Special issue to mark the retirement of Professor of Martin Vessey. PMID- 11417061 TI - Ulcer complications associated with anti-inflammatory drug use. What is the extent of the disease burden? AB - Information on the intake of non-steroidal anti-inflammatory drugs (NSAIDs) and on aspirin taken regularly by patients with peptic ulcer bleeding aged 60 years and over was used in conjunction with data measuring the overall frequency of hospital admissions with ulcer bleeding in England and Wales to determine the annual burden of disease imposed by particular treatment strategies. Over 40% of the calculated 8528 episodes of ulcer bleeding in those aged 60 years and over, and over 40% of the estimated 981 deaths each year would seem to be causally related to the treatments. Substitution of the NSAID with the lowest associated risk would be expected to reduce the frequency of non-aspirin NSAID-associated episodes of ulcer bleeding, and deaths, each by over 70%. Use of the lowest conventional dose of regular prophylactic aspirin (75 mg) would also be expected to reduce the frequency of treatment-related episodes, and deaths, by nearly 30%. Both strategies employed together would be expected to reduce NSAID and regular aspirin-related bleeding ulcer admissions from 4121 to less than 2184, and deaths from 523 to less than 250. Substitution of completely safe anti-inflammatory analgesics and anti-platelet drugs would be expected to reduce admissions from 4121 to 1072, and deaths from 523 to 123. PMID- 11417062 TI - Contraception--past, present and future: a presentation given at the symposium to honour the retirement of Professor Martin Vessey. PMID- 11417063 TI - Developments in the management of menopause and hormone replacement therapy: a presentation given at the symposium to honour the retirement of Professor Martin Vessey. PMID- 11417064 TI - HPV testing in cervical screening. PMID- 11417065 TI - The MRC years: presented at the symposium for Professor Martin Vessey. PMID- 11417066 TI - Martin Vessey: departmental and divisional perspective. PMID- 11417067 TI - A festschrift for Martin Vessey: pioneering academic and NHS links. PMID- 11417068 TI - Evolution of division of public health and primary health care: a regional health authority viewpoint. PMID- 11417069 TI - The evolution of screening. AB - Botany is usually considered to be the gentlest of sciences with botanists being regarded as people who study relatively safe specimens, compared with, for example, anthropologists or microbiologists. However, botanists have their moments, particularly when collecting new species. The great botanists of the eighteenth and nineteenth centuries risked their lives in collecting and bringing back species, which we now take for granted, and Robert Brown was one of these adventurers, a young Scot who accompanied Sir Joseph Banks to New Holland. It was not, however, for his adventurous lifestyle that Brown is remembered but for his startling observation of the movements of pollen grains on a microscope slide. He noted that the pollen grains were in perpetual agitated motion, without purpose or direction but full of energy. This motion, called Brownian motion, arises from the movement of molecules, and Brownian motion is the term that has been applied to much of healthcare, including many screening programmes, which have in the past been marked more by the amount of energy and activity than by a clear sense of direction or positive achievement. PMID- 11417070 TI - The role of cohort studies in medical research. PMID- 11417071 TI - Improving family ties: an examination of the complementary disciplines of pharmacoepidemiology and clinical pharmacology. PMID- 11417072 TI - Pharmacoepidemiology and drug safety. PMID- 11417073 TI - [Equine motor neuron disease: a review based on a case report]. AB - A 10-year-old, non-pregnant Dutch Warmblood mare was referred to the Department of Equine Science because of chronic weight loss, despite good appetite, and dullness. Clinical examination revealed muscle atrophy, trembling of the limb muscles, an abnormal stance in which all four limbs were placed under the body, and an abnormal low head carriage. The plasma vitamin E concentration was markedly decreased (0.2 mumol/l), the electromyographic (EMG) examination was consistent with denervation, and the oral glucose absorption test was below the reference value (40% increase over the resting glucose level). Because of the clinical diagnosis of equine motor neuron disease (EMND), the horse was euthanazed and post-mortem examination confirmed this diagnosis. Based on the similarity in pathological findings, EMND can be compared to amyotrophic lateral sclerosis (ALS) in humans. However, in horses, only the lower motor neurons and occasionally some nuclei of the cranial nerves are affected. Because of the low plasma vitamin E concentration found in horses with EMND, an absolute or a relative antioxidant deficiency can be involved in the pathogenesis. In general, it is a progressive disease process and stabilization of the situation is the best feasible result. There is no specific therapy other than vitamin E supplementation and the prognosis is poor. PMID- 11417074 TI - [Veterinarians as experts in court cases]. PMID- 11417075 TI - [Predecessors: veterinarians from earlier times. (43). Henri Marie Bouley (1814 1886)]. PMID- 11417076 TI - ['If you want something, just do it!']. PMID- 11417077 TI - [Interaction of diet and veterinary drugs: 'Take it easy, but don't forget it!']. PMID- 11417078 TI - [Who is the bitten dog? Legal liabilities for damage by animals]. PMID- 11417079 TI - [Legal cases, where are the boundaries?]. PMID- 11417080 TI - [Members of the KNMvD are boring, slow and old-fashioned]. PMID- 11417081 TI - [Appreciation of membership in KNMvD]. PMID- 11417082 TI - [Together we are strong!?]. PMID- 11417083 TI - ['Tomorrow we start vaccination, that is our duty']. PMID- 11417084 TI - Leave a legacy. PMID- 11417085 TI - Pediatric pharmacology. What you need to know for the next pediatric call. PMID- 11417086 TI - Bringing young people into EMS. PMID- 11417087 TI - Managing risk in EMS. PMID- 11417088 TI - Assessment of the stroke patient using the NIH stroke scale. AB - The NIH scale requires no advanced expertise to complete. Any level of EMS provider can use the form if given proper orientation and training, with an in service to explain the form and clear up any ambiguous language in it. Scenario based practice sessions following a short class are ideal. A nurse from a hospital's stroke team would be the best person to teach the class, since he/she would be intimately familiar with the form and how to use it. The NIHSS is the ideal assessment tool to make prehospital examination of stroke patients more aggressive. Although ideal, the NIHSS should only be used when transport time allows. By no means should a stroke patient's transport be delayed to administer the scale. If conditions and time do not allow, the provider should revert back to the CPSS as an assessment tool. If transport time allows, however, the NIHSS is the proper exam to use. It is aggressive, simple to use and cases the transition of the patient from ambulance to emergency department. It also saves time for the patient by allowing the doctor to provide proper treatment more quickly. The forms are free to copy from the NIHSS and, therefore, costs are minimal to implement its use. Any solution that assists patient, physician and provider in an aggressive yet economical manner needs to be implemented. The positive aspects of the NIHSS in prehospital care span across the board and will be realized in patient care. PMID- 11417089 TI - Living in the shadow of PTSD. PMID- 11417090 TI - Family-centered prehospital care. AB - Family-centered care is mutually beneficial collaboration between patients, family members and healthcare professionals to improve the overall health and safety of the community. Because of their frequent presence at prehospital emergency scenes, family members often participate in prehospital care prior to the arrival of EMS. EMS professionals generally have little or no training in managing family members or in integrating their needs into the needs of their patients. Many EMS professionals believe interacting with family members is a matter of common sense, but our multicultural environment and advances in medical technology make the issue more complex. EMS professionals favor most family centered care concepts, but are hesitant to open their patient compartments or their advisory boards to family members. For more information about family centered prehospital care, visit NAEMT's website at www.naemt.org. PMID- 11417091 TI - Personal malpractice insurance. PMID- 11417092 TI - A history of EMS in the Dominican Republic. PMID- 11417093 TI - What are you prepared to do? PMID- 11417094 TI - Treating patients with autism. PMID- 11417095 TI - Human resource training: responding to Supreme Court's direction. PMID- 11417096 TI - Is this really a nursing home? Innovative models focus on enhancing quality of life. PMID- 11417097 TI - Ingenuity encounters incontinence. PMID- 11417098 TI - Our annual tribute to people who make a difference. PMID- 11417099 TI - Medicare decision reversed. PMID- 11417100 TI - Who's watching the wanderers? PMID- 11417101 TI - Increasing safety, managing risk. AB - Before criminal background checks became standard industry practice, a long term care administrator in Illinois paid $15 per inquiry for such checks on potential hires for his facility. Management ordered the inquiries halted for budgetary reasons. A few months later, facing holiday staff shortages around Christmastime, the administrator hired two new aides to help round out a skeletal staff. "I did the registry checks, the reference checks, everything came in fine," recalls the administrator, who asked to remain anonymous in our reportage. "I came back in on Monday--Christmas was on Sunday--and got a report from [a resident's] family members that their mother had been sexually abused Christmas night." A year later, in legal proceedings over the incident, the administrator was summoned to give a deposition about the center's hiring practices. In fact, it had been discovered that one of the aides hired did have a record of abuse. Because the administrator cited that he was not permitted to perform criminal background checks, a decision with which he disagreed, he became a key witness for the plaintiff. Management admitted liability and paid $1 million in damages awarded the plaintiff. PMID- 11417102 TI - Safe passages. Lose the institutional look without sacrificing safety. PMID- 11417103 TI - Who's monitoring resident health? PMID- 11417104 TI - PDA letter to the FDA regarding process simulation testing. PMID- 11417105 TI - Application of recovery tests in the validation of immunoassays for assessing the immunogenicity of B. anthracis PA vaccine. AB - In the quantitative assessment of polyclonal serum antibodies, the complex composition and characteristics of the analyte population (serum antibodies) restricts the capability of constructing appropriately defined calibration standards. This fact limits the application of the conservative recovery tests to the validation of immunoassays aimed at determining serum antibody levels. The present report describes a modification of recovery tests that overcomes this impediment. The modified approach is based on a dilution analysis system, where a given immune serum is serially diluted in normal serum and the antibody titers in each of the derived diluted samples are then determined. Expected sample titers (calculated on the basis of the relevant dilution factors) are plotted against the respective observed results, and the resulting recovery curve is then examined by means of a regression analysis, according to the standard rules of the conservative recovery analysis. This approach was tried with two immunoassay systems, Enzyme Linked Immunosorbent Assay (ELISA) and Neutralizing Antibodies (NtAb) immunoassays, aimed at assessing the immunogenicity in guinea pigs of B. anthracis protective antigen (PA) vaccine. In a series of feasibility studies using a recovery simulation model (dilutions made in the immunoassay diluent, rather than in normal serum) the average recovery levels in ELISA and NtAb immunoassays were 0.99 +/- 0.011 and 1.02 +/- 0.04 respectively, and the 99% confidence intervals contained the target 100% value. Regression lines were proved to be linear demonstrating R > 0.97 in all cases. The 99% confidence intervals around the observed slopes and intercepts always contained the corresponding target values 1 and 0. The relative standard deviation (RSD) in the ELISA and NtAb immunoassays was found to be 0.01 and 0.025 respectively. All of the above experimental results were not affected by the serum antibody titer, or by day-to-day variations embodied in these immunoassay systems. When true recovery tests were applied to the above immunoassays, essentially identical results were obtained. In both assays the correlation coefficients were in the range of 0.96-1, recoveries were found to be in the range of 0.90-1.06, and RSD values were in the range of 0.02-0.025. All the recovery deviations from the target value of 1 were not statistically significant. The hitherto observed experimental findings illustrate the capability of the dilution analysis system to allow the application of recovery tests to the validation of quantitative immunoassays, which are based on the procedure of serum titrations. PMID- 11417106 TI - Determination of moisture in rubber stoppers: effect of Karl Fischer oven temperatures. AB - Moisture released from rubber stoppers of pharmaceutical products may adversely affect product quality during storage. The objective of the current study was to identify an appropriate method to quantify free moisture in processed stoppers. The moisture content in stoppers is commonly determined by the Karl Fischer (KF) method. However, for the FM257/2 stoppers from Helvoet, our results suggest that the KF assay results are always lower than that from the gravimetric method and increase with the KF oven temperatures used. The systematic study presented in this manuscript shows that, although moisture results vary with respect to KF oven temperatures, reproducible results are obtained at fixed temperatures. Therefore, the observed variation can not be explained by the release of volatile chemicals from this rubber-affecting KF reagents. Furthermore, by extrapolating moisture results obtained with different thicknesses of rubber slabs to zero thickness, it was found that the diffusion barrier is also not a major source for the variation. It is hypothesized that the moisture in rubber stoppers can be free or bound. The KF method measures both free and bound moisture. As higher KF oven temperatures are used, more bound moisture is released and determined by the KF method. By deducting the bound moisture (measured by the KF method for samples dried at 100 degrees C for one week) from the sample moisture, the amount of free moisture (which is releasable) can be estimated. A KF oven temperature at 250 degrees C is recommended based on the accuracy and precision of the results. This method was validated by showing consistency with the gravimetric method for stoppers undergoing desorption at 25 degrees C and 0% relative humidity (equilibrated with P2O5). An example is also presented to show the application of this method. PMID- 11417107 TI - Setting threshold limits for the significance of objectionable microorganisms in oral pharmaceutical products. AB - Due to the increasing number of immuno-compromised patients, increased attention is paid to the quantitation and identification of microorganisms in oral pharmaceutical products; therefore, a systematic approach is required by the manufacturers of non-sterile oral pharmaceuticals to evaluate the significance of microbial isolates other than primary pathogens and/or those in the product specification based on the number of organisms present, the type of dosage form, and the potential hazard to the user. Limits for objectionable microorganisms in oral products intended for use by immuno-compromised patient populations such as pediatric, HIV, cancer, etc., must be tighter than the limits for oral products intended for treating patients with diseases or conditions not affecting their immune systems because patients with deficient immune systems are more at risk of microbial infections. Smaller numbers of opportunistic pathogens become infectious when resistance mechanisms are impaired, either by severe underlying disease, or by use of immunosuppressive drugs. This article proposes a systematic approach for evaluating the significance of microbial isolates other than primary pathogens and/or those in the specification (e.g., Staphylococcus aureus and Pseudomonas aeruginosa) of non-sterile oral pharmaceutical products by setting appropriate threshold limits. PMID- 11417108 TI - Environmental monitoring: myths and misapplications. PMID- 11417109 TI - Environmental monitoring: misconceptions and misapplications. PMID- 11417110 TI - Synthetic macromolecular drug carriers: biodistribution of poly[(N-2 hydroxypropyl)methacrylamide] copolymers and their accumulation in solid rat tumors. AB - To optimize polymer design for tumor directed drug delivery, the fate and the total body distribution of soluble synthetic macromolecules, derived from copolymers of [(N-2-(hydroxypropyl)methacrylamide] (HPMA) were monitored scintigraphically after radiolabeling with 131I during a seven day time window. Equimolar concentrations of radioiodinated copolymers of HPMA with small amounts of methacryloyltyrosinamide (pHPMA) differing in molecular weight (23.4 kD, 27.3 kD, 30.5 kD, 44 kD, 58.4 kD, 60.1 kD) were injected intravenously into Copenhagen rats bearing Dunning prostate carcinomas (subline R3327-AT1). Scintigraphic data were validated by determining absolute amounts of [131I]pHPMA in both tumor tissue and normal organs after sacrificing the animals. Copolymers were cleared from blood circulation in a molecular-weight dependent manner, either via excretion or by extravasation into normal and neoplastic tissues. While distribution patterns for pHPMAs in normal organs were quite similar, absolute amounts of copolymer uptake differed. The higher the molecular weight, the more radioactivity was taken up by the organs. Highest amounts of radioactivity were seen in the lung, liver, and spleen. In solid tumors, kinetics of pHPMA accumulation was clearly dependent on molecular weight. pHPMAs below the renal threshold peaked at 24 hours p.i. and then remained constant. In contrast, copolymers above the renal clearance threshold displayed a continuous accumulation reaching a significantly higher tumor uptake, presumably due to the very small or non existent polymer release from tumor tissue. Absolute amounts of tumor uptake determined by dissection analysis were 0.5 +/- 0.1% of injected dose/g tissue for the 27.3 kD pHPMA and 1.2 +/- 0.1% for the 60.1 kD pHPMA, respectively. In conclusion, our results demonstrate the influence of the molecular weight of the synthetic polymer pHPMA on plasma circulation time, excretion and organ clearance. While pHPMAs are cleared from all normal tissues except the spleen quite effectively, these polymers accumulate in solid tumors in a size dependent manner, due to the well known "enhanced permeability and retention" (EPR) effect. These data are of fundamental interest for ongoing studies on the pharmacokinetics of synthetic polymers, especially when these molecules are conjugated with targeting moieties and therapeutic or diagnostic agents. PMID- 11417111 TI - Alternative medicine: herbal drugs and their critical appraisal--Part I. AB - Among alternative therapeutic approaches that have shown global popularity during the past decades, herbal medicine stands out as a major concern in the countries where allopathic medicine prevails. The sales of herbal products as health care adjuvants in these countries have increased exponentially. Lack of quality control, commercial profiteering and exploitation leading to adulterations, lack of proper knowledge about the herbs and their contents that may exhibit drug-drug interactions and other adverse side-effects, and inappropriate usage of the herbal products have become a cause for concern in the health care professions, particularly in the United States. This review provides an incisive description of the known chemical, pharmacological, clinical and toxicological profiles of four of the most widely used herbal products. PMID- 11417112 TI - Quantitative structure-activity relationships of antianginal drugs. AB - Quantitative structure-activity relationships (QSARs) of various classes of antianginal drugs, e.g. nitrates, beta-adrenergic blocking agents (beta blockers), and calcium channel blockers (calcium antagonists), have been reviewed. This review gives an overall picture of the mode of action of each class of drugs and points out the specific physicochemical and structural properties that govern their activity. It is observed that in almost all kinds of antianginal drugs the lipophilic factor plays an important role and the next important factor seems to be the steric ones. The electronic factors are found to be occasionally important. In the case of beta-blockers, the most common factor that appeared to govern the activity remained the lipophilicity. In nitrates, too, the activity is observed to primarily depend upon the lipophilicity. In calcium channel blockers, however, the dominant effect is seen to be of steric factors. The steric roles may be essential in drug-receptor interactions, which seem to involve both hydrophobic, and to a lesser extent, electronic interactions. PMID- 11417113 TI - Purine analogs as CDK enzyme inhibitory agents: a survey and QSAR analysis. AB - Characterization of the cell cycle has introduced CDKs and other proteins as possible targets for inhibition of cell proliferation, such as, CDK1 and CDK2, whose inhibition may be useful in the treatment of proliferative disorders. Structure-activity analyses have been instrumental in the design and discovery of potent CDK inhibitors, such as purine analogs, which have increased in potency from the micromolar to the nanomolar level. X-ray crystallography and molecular modeling have provided evidence that these compounds act on the CDK target enzyme. Selected CDK inhibitors have successfully entered clinical trials. Further characterization of the cell cycle to identify molecular targets to inhibit cell proliferation, QSAR and SAR studies, and clinical trials may expedite the development of CDK inhibitors for therapeutic use. The ultimate goal of these studies is to determine whether specific CDKs, CDK1 or CDK2, are enzymes essential to cell proliferation that can be targeted for treatment of proliferative disorders. CDK1 and CDK2 are viable molecular targets for cancer therapies based on isolated-enzyme inhibition by CDK inhibitors, successful clinical trials of CDK1 and CDK2 inhibitors, and x-ray crystallographic confirmation of CDK inhibitors binding to the putative target enzyme active site. It is now reported that CDK1 inhibitory activities of purine analogs correlate with the physiochemical parameters of purine analogs. Enzyme inhibition [1-5], clinical trials (see Tab. 1), x-ray crystallographic [4, 6, 7] and QSAR correlation studies are evidence that specific CDK1 and/or CDK2 inhibitors are potentially useful agents for various cell-proliferation disorders. A brief overview of the cell cycle precedes a literature review of clinical applications of CDK inhibitors, followed by a new QSAR study, and a SAR and molecular modeling discussion. PMID- 11417114 TI - Anti-influenza drugs and neuraminidase inhibitors. AB - Each year, influenza viruses are responsible for considerable illness, complications and mortality. An effective treatment will have a major impact on the severe personal and economic burden that this disease incurs. There are several points in the influenza life cycle that may be potentially inhibited. One critical point is the release of newly synthesized virions from the host cell surface. Viral neuraminidase (NA) cleaves the virus from host cell sialic acid residues allowing infection of other host cells. Rationally designed NA inhibitors that block the viral life cycle are now in the clinic and these molecules are effective and safe for the treatment of influenza. Compared with other anti-influenza agents the NA inhibitors are well tolerated, effective against all influenza types and there has been little evidence of the emergence of viral resistance. NA inhibitors provide an important new therapeutic weapon for the management of influenza infection. PMID- 11417115 TI - Current and potential therapies for the treatment of herpesvirus infections. AB - Human herpesviruses are found worldwide and are among the most frequent causes of viral infections in immunocompetent as well as in immunocompromised patients. During the past decade and a half a better understanding of the replication and disease causing state of herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), varicella-zoster virus (VZV), and human cytomegalovirus (HCMV) has been achieved due in part to the development of potent antiviral compounds that target these viruses. While some of these antiviral therapies are considered safe and efficacious (acyclovir, penciclovir), some have toxicities associated with them (ganciclovir and foscarnet). In addition, the increased and prolonged use of these compounds in the clinical setting, especially for the treatment of immunocompromised patients, has led to the emergence of viral resistance against most of these drugs. While resistance is not a serious issue for immunocompetent individuals, it is a real concern for immunocompromised patients, especially those with AIDS and the ones that have undergone organ transplantation. All the currently approved treatments target the viral DNA polymerase. It is clear that new drugs that are more efficacious than the present ones, are not toxic, and target a different viral function would be of great use especially for immunocompromised patients. Here, we provide an overview of the diseases caused by the herpesviruses as well as the replication strategy of the better studied members of this family for which treatments are available. We also discuss the various drugs that have been approved for the treatment of some herpesviruses in terms of structure, mechanism of action, and development of resistance. Finally, we present a discussion of viral targets other than the DNA polymerase, for which new antiviral compounds are being considered. PMID- 11417116 TI - New results on the chemistry of lichen substances. PMID- 11417117 TI - Pore-forming bacterial protein toxins: an overview. PMID- 11417118 TI - Helicobacter pylori vacuolating cytotoxin: cell intoxication and anion-specific channel activity. PMID- 11417119 TI - Pore-forming colicins and their relatives. AB - The pore-forming colicins, the first proteins that were capable of forming voltage-dependent ion channels to be sequenced, have turned out to be both less tractable and more mysterious than imagined; yet they have proved interesting at every step of their short journey from producing cell to vanquished target cell. Starting out as a remarkably extended water-soluble protein, the colicin molecule is designed to interact simultaneously with several components of the complex membrane of the target cell, transform itself into a membrane protein, and become an ion channel with inscrutable properties. Unraveling how it does all this appears to be leading us into the dark recesses of protein/protein and protein/membrane interaction, where lurk fundamental processes reluctantly waiting to be revealed. PMID- 11417120 TI - The cholesterol-dependent cytolysins. AB - In view of the recent studies on the CDCs, a reasonable schematic of the stages leading to membrane insertion of the CDCs can be assembled. As shown in Fig. 3, we propose that the CDC first binds to the membrane as a monomer. These monomers then diffuse laterally on the membrane surface to encounter other monomers or incomplete oligomeric complexes. Presumably, once the requisite oligomer size is reached, the prepore complex is converted into the pore complex and a large membrane channel is formed. During the conversion of the prepore complex to the pore complex, we predict that the TMHs of the subunits in the prepore complex insert into the bilayer in a concerted fashion to form the large transmembrane beta-barrel, although this still remains to be confirmed experimentally. Many intriguing problems concerning the cytolytic mechanism of the CDCs remain unsolved. The nature of the initial interaction of the CDC monomer with the membrane is currently one of the most controversial questions concerning the CDC mechanism. Is cholesterol involved in this interaction, as previously assumed, or do specific receptors exist for these toxins that remain to be discovered? Also, the trigger for membrane insertion and the regions of these toxins that facilitate the [figure: see text] interaction of the monomers during prepore complex formation are unknown. In addition, the temporal sequence of the multiple structural changes that accompany the conversion of the soluble CDC monomer into a membrane-inserted oligomer have yet to be defined or characterized kinetically. PMID- 11417121 TI - Aerolysin from Aeromonas hydrophila and related toxins. PMID- 11417122 TI - Staphylococcal pore-forming toxins. PMID- 11417123 TI - RTX toxin structure and function: a story of numerous anomalies and few analogies in toxin biology. AB - It can be agreed that RTX toxins contribute to the pathogenesis of different diseases by causing dysfunction of the general cellular reactions of the immune response. The suggestion that RTX toxins induce cytokine production in nonimmune cells that would ultimately cause tissue damage is an expansion of their role in disease pathogenesis (Uhlen et al. 2000). Investigators in the RTX toxin field may not agree with me, but precise and satisfactory answers to the following questions are not yet available. How do RTX toxins mechanistically damage a cell? Do RTX toxins have receptors in the classic sense, in which there is a reversible ligand and receptor complex? What is responsible for the common Ca2+ ion influx in affected cells? The recent observation that an RTX toxin stimulates host-cell mediated Ca2+ ion oscillation in part challenges the long held concept that these toxins damage cells by the direct formation of pores. Are the Ca2+ ion fluxes truly the noxious cellular insult? What is the final molecular structure of RTX toxins at the time they cause cellular death? How does the common requirement for acyl modification among RTX toxins fit into the toxin structure and mechanism of cellular killing, particularly when mixtures of unusual fatty acids are used by some toxins? There are a number of outstanding laboratories throughout the world that are seeking answers to these questions. We can reasonably expect that during the next decade research on the structure and function of RTX toxins will lead to new chemotherapeutic targets and reagents for basic cell biology and biotechnology. PMID- 11417124 TI - Methods and assays to investigate nuclear export. PMID- 11417125 TI - Nuclear export of herpes virus RNA. PMID- 11417126 TI - Nuclear export of adenovirus RNA. PMID- 11417127 TI - Nuclear export mediated by the Rev/Rex class of retroviral Trans-activator proteins. PMID- 11417128 TI - Constitutive transport element-mediated nuclear export. PMID- 11417129 TI - Nuclear pore complex architecture and functional dynamics. PMID- 11417130 TI - The concept of neurotropism and selective vulnerability ("pathoclisis") in virus infections of the nervous system--a historical overview. PMID- 11417131 TI - Interactions of viral proteins with neurotransmitter receptors may protect or destroy neurons. PMID- 11417132 TI - Mechanisms of virus-induced neuronal damage and the clearance of viruses from the CNS. PMID- 11417133 TI - The mechanisms of direct, virus-induced destruction of neurons. PMID- 11417134 TI - Borna disease virus infection of adult and neonatal rats: models for neuropsychiatric disease. AB - Animal models provide unique opportunities to explore interactions between host and environment. Two models have been established based on Borna disease virus infection that provide new insights into mechanisms by which neurotropic agents and/or immune factors may impact developing or mature CNS circuitry to effect complex disturbances in movement and behavior. Note in press: Since this chapter was submitted, several manuscripts have been published that extend findings reported here and support the relevance of BDV infections of neonatal Lewis rats as models for investigating mechanisms of neurodevelopmental damage in autism. Behavioral abnormalities, including disturbed play behavior and chronic emotional overactivity, have been described by Pletnikov et al. (1999); inhibition of responses to novel stimuli were described by Hornig et al. (1999); loss of Purkinje cells following neonatal BDV infection has been demonstrated by Eisenman et al. (1999), Hornig et al. (1999), and Weissenbock et al. (2000); and alterations in cytokine gene expression have been reported by Hornig et al. (1999), Plata-Salaman et al. (1999) and Sauder et al. (1999). PMID- 11417135 TI - The mechanisms of neuronal damage in retroviral infections of the nervous system. PMID- 11417136 TI - Prion-induced neuronal damage--the mechanisms of neuronal destruction in the subacute spongiform encephalopathies. AB - Prion diseases are characterized by the accumulation of a specific disease associated isoform of the prion protein (PrP), termed PrPSc, which is the main, if not the only, component of the infectious agent termed prion. PrPSc is derived by an autocatalytic post-translational process involving conformational changes from the normal host-encoded isoform of the prion protein, termed PrPC. PrPC is a copper-binding glycoprotein attached to the cell membrane of neurons and other cells by means of a GPI anchor. The pattern of neurodegeneration differs between variants of prion disease and is related to the pattern of PrPSc deposition and differences in susceptibility of different cell types to the disease process. The pattern of PrPSc deposition depends on the strain of the agent and the PrP genotype of the host. Strain properties of prions appear to be related to different pathological conformations of PrPSc. Neuronal cell death is a salient feature in the pathology of prion diseases. Histological and electron microscopical studies have shown that cell death in prion disease occurs by apoptosis. Apoptosis of neuronal cells can also be induced in vitro by exposure to PrPSc or a neurotoxic peptide fragment corresponding to amino acids 106-126 of human prion protein (PrP106-126). Both in vitro and in vivo, the toxicity of PrPSc and PrP fragments appears to depend on neuronal expression of PrPC and on microglial activation. Activated microglial cells release pro-inflammatory cytokines and reactive oxygen species. Cell culture experiments suggest an important role of microglia-mediated oxidative stress in the induction of neuronal cell death. Only limited data are available on direct effects of PrPSc on neuronal cells. Potential effects include increased formation of an aberrant transmembrane form of PrP, termed CtmPrP, and changes in plasma membrane properties. In addition to direct and indirect toxic effects of PrPSc, a loss of function of PrPC may contribute to neuronal cell death. Potential mechanisms include disturbances in cerebral copper metabolism and antioxidative defense mechanisms. A better understanding of the pathogenesis of neuronal cell death in prion diseases may also have important therapeutic implications in the future. PMID- 11417137 TI - The role of T-cell-mediated mechanisms in virus infections of the nervous system. AB - T lymphocytes play a decisive role in the course and clinical outcome of viral CNS infection. Summarizing the information presented in this review, the following sequence of events might occur during acute virus infection: After invasion of the host and a few initial rounds of replication, the virus reaches the CNS in most cases by hematogeneous spread. After passage through the BBB, CNS cells are infected and replication of virus in brain cells causes activation of the surrounding microglia population. Moreover, local production of IFN alpha/beta induces expression of MHC antigens on CNS cells, and microglial cells start to phagocytose cellular debris, which accumulates as a result of virus induced cytopathogenic effects. Upon phagocytosis, microglia becomes more activated; they up-regulate MHC molecules, acquire antigen presentation capabilities and secrete chemokines. This will initiate up-regulation of adhesion molecules on adjacent endothelial cells of the BBB. Transmigration of activated T lymphocytes through the BBB is followed by interaction with APC, presenting the appropriate peptides in the context of MHC antigens. It appears that CD8+ T lymphocytes are amongst the first mononuclear cells to arrive at the infected tissue. Without a doubt, their induction and attraction is deeply influenced by natural killer cells, which, after virus infection, secrete IFN-gamma, a cytokine that stimulates CD8+ T cells and diverts the immune response to a TH1-type CD4+ T cell-dominated response. Following the CD8+ T lymphocytes, tissue-penetrating, TH1 CD4+ T cells contact local APC. This results in a tremendous up-regulation of MHC molecules and secretion of more chemotactic and toxic substances. Consequently an increasing number of inflammatory cells, including macrophages/microglia and finally antibody-secreting plasma cells, are attracted to the site of virus infection. All trapped cells are mainly terminally differentiated cells that are going to enter apoptosis during or shortly after exerting their effector functions. The clinical consequences and the influence of the effector phase on the further course of the infection depends on the balance and fine-tuning of the contributing lymphoid cell populations. Generally, any delay in the recruitment of effector lymphocytes to the tissue or an unbalanced combination of lymphocyte subsets allows the virus to spread in the CNS, which in turn will cause severe immune-mediated tissue effects as well as disease. If either too late or partially deficient, the immune system response may contribute to a lethal outcome or cause autosensitization to brain-specific antigens by epitope spreading to the antigen-presenting system in peripheral lymphoid tissue. This could form the basis for subsequent booster reactions of autosensitized CD4+ T cells--a process that finally will end in an inflammatory autoimmune reaction, which in humans we call multiple sclerosis. In contrast, a rapid and specific local response in the brain tissue will result in efficient limitation of viral spread and thereby a subclinical immune system-mediated termination of the infection. After clearance of virus-infected cells, downsizing of the local response probably occurs via self-elimination of the contributing T cell populations and/or by so far unidentified signal pathways. However, much of this is highly speculative, and more data have to be collected to make decisive conclusions regarding this matter. Several strategies have been developed by viruses to escape T cell-mediated eradication, including interference with the MHC class I presentation pathway of the host cell or "hiding" in cells which lack MHC class I expression. This may result in life-long persistence of the virus in the brain, a state which probably is actively controlled by T lymphocytes. Under severe immunosuppression, however, reactivation of viral replication can occur, which is a lethal threat to the host. PMID- 11417138 TI - Virus-induced autoimmune reactions in the CNS. PMID- 11417139 TI - Slow and persistent virus infections of neurones--a compromise for neuronal survival. PMID- 11417140 TI - Neuronal latency in human and animal herpesvirus infections. PMID- 11417141 TI - Programmed cell death in virus infections of the nervous system. PMID- 11417142 TI - Structure and function of the bladder neck. PMID- 11417143 TI - Efficacy of induction and difficulty level in durability of post-hypnotic suggestions. AB - We examined whether participants instructed to reenter a hypnotic state as part of the post-hypnotic suggestion (PHS) show less decay in responding over an 8 week period than participants who do not receive such instructions. We also attempted to replicate Trussell, Kurtz, and Strube's (1996) finding on impact of difficulty level of suggestion on response curve. Fifty-nine highly susceptible participants were selected by the Stanford Hypnotic Susceptibility Scale: Form C (SHSS:C) and were assigned to one of four groups (two levels of Difficulty [easy hard] x two levels of Condition [hypnotic PHS, non-hypnotic PHS]). Participants were tested for PHS at 1, 3, 6, and 8 weeks. A 2 x 2 x 4 (Difficulty x Condition x Time) factorial ANOVA was conducted, with Time as a repeated-measure. The outcome variable at each time was either pass or fail for relevant suggestion. None of the effects containing Condition as a term were significant indicating there is no advantage to using Berrigan, Kurtz, Stabile, and Strube's (1991) atypical induction technique to influence the durability of PHS. We found a significant Time effect but failed to replicate Trussell et al.'s findings for Difficulty level. The differing results found in these three recent studies (Berrigan et al., Trussell et al., and the current study) suggest the effects for durability of PHS may be quite fragile in spite of rigorous experimental controls used in all three studies. PMID- 11417144 TI - Automatic word processing: a new forum for hypnotic expression. AB - Automatic word processing, a form of dissociation utilizing hypnosis and computer technology, was developed to provide patients a forum for hypnotic expression with unique features. Using automatic word processing, the six patients in this report were able to express themselves extensively, edit their responses to queries, and even write poetry while doing hypnosis. Automatic word processing can be used with some patients to gain important insights and therapeutic successes that might not be achieved as easily by other means. Because this is a new technique, much of its potential remains to be defined. PMID- 11417145 TI - Rapid self-hypnosis: a new self-hypnosis method and its comparison with the Hypnotic Induction Profile (HIP). AB - Despite its clinical importance, there are few systematic studies on the application of self-hypnosis. Rapid Self-Hypnosis (RSH) was created to provide a new procedure that is easy, comfortable, fosters alertness, and can be done covertly in everyday life. We present it as an alternative to the self-hypnosis version of the Hypnosis Induction Profile (HIP). Using a crossover design, we found in an experimental session that the RSH and the HIP produced comparable objective and subjective scores in the Barber Suggestibility Scale (BSS). However, as compared with the HIP, participants rated RSH as significantly more coherent, pleasant, faster and easier to learn, more likely to be used in everyday life and go unnoticed by others, less bothersome to use, and more likely to be used in private. Additional research should clarify whether these differences are reliable and have clinical significance. Our results suggest that RSH will be a valuable addition to the clinician's arsenal. PMID- 11417146 TI - Freud's relevance to hypnosis: a reevaluation. AB - In this paper we examine Freud's life and thinking, based on his collected works, and reevaluate some of his ideas in the light of various aspects of contemporary hypnosis research. Although Freud has often been blamed for simplistic thinking about hypnosis and for its eclipse during the opening decades of this century, his writings reveal a rich theory of hypnosis and a frank acknowledgement of the debt psychoanalytic theory and practice owe to it. Even though he abandoned hypnosis as a clinical tool, Freud maintained a theoretical interest in the subject and in many respects anticipated issues in current research. Whereas his emphasis on the hypnotist's skill may have been exaggerated, his insights concerning attention, social expectations, group dynamics, reality testing, and the relationship between hypnosis and sleep have been borne out by empirical investigations. PMID- 11417147 TI - Perceptual reconstruction in the treatment of inordinate grief. AB - Troubled by continuing emotionally painful memories of her ill mother's face as she struggled to breathe during her terminal days in the hospital, an adult daughter requested hypnosis to "erase" those images from her memory. Since that is not feasible, the subject instead was provided in a single session with a hypnosis-based, imagery-focused strategy in which the disturbing images of her mother were replaced by hypnotically reconstructed positive memories as she had appeared in prior, happy times. This was successful: the restructured images were maintained and grew stronger over time as reported four months post-session. It was concluded that hypnotically restructured memories can be useful techniques in the management of inordinate grief. PMID- 11417148 TI - Dyspepsia as a somatic expression of guilt: a case report. AB - A 58-year-old woman developed chronic, severe symptoms of heartburn, epigastric pain, and regurgitation that persisted for 2 years. She underwent a thorough evaluation and no organic cause was identified. Therefore, a diagnosis of dyspepsia was made. Her symptoms were refractory to pharmacological treatment. Upon further probing, the patient reported that the onset of her symptoms coincided with the death of her son of cancer 2 years earlier. She blamed herself for the death of her son and admitted to a need for self-punishment. A brief course of treatment using metaphors and hypnosis resulted in a complete resolution of her symptoms, which did not recur during a follow-up of 12 years. This is the first published report of the treatment of dyspepsia using hypnotic methods. PMID- 11417149 TI - Hypnotherapy in adolescents with trichotillomania: three cases. AB - Trichotillomania is not rare in adolescence. Psychotherapy is often ineffective, and cognitive behavioral therapy in combination with serotonin-specific reuptake inhibitors seems to be the treatment of choice. Some cases are resistant to all therapy. This paper reports on three adolescents with pure trichotillomania who responded to the imaginative hypnotherapy technique with Ericksonian suggestions. The patients described their hair as weak and vulnerable and needy of protection. In therapy, the patient was assigned the role of "patron of the hair" thereby giving him/her control of the situation. Hair pulling was significantly reduced, and the improvement was sustained throughout the 6-month follow-up. These cases suggest that imaginative techniques may be effective in adolescents with trichotillomania. Further controlled studies in adolescent population are needed to confirm this assumption. PMID- 11417150 TI - The response set theory of hypnosis: expectancy and physiology. AB - A recent exposition of the response set theory of hypnosis (Kirsch, 2000) contained incorrect and misleading figures. The correct figures illustrated a complementary relation between mental and physiological phenomena. The figures as published erroneously suggested that the author espoused epiphenomenalism. As shown in this corrected version, Kirsch proposes that mind states and body states be considered as two ways of viewing a single psychophysiological phenomenon. PMID- 11417151 TI - Custom foot orthoses for running. AB - This article reviews the epidemiology of running injuries, the rationale and application of custom foot orthoses for running, and the design and material choices for fabrication in one laboratory. In an age of evidence-based medicine, much research must be done to look further at foot orthotic efficacy and how these devices contribute to an injury management plan. When designing custom foot orthoses for running, the differences between walking and running mechanics must be considered. Choices of the foot care specialist depend on patient body habitus, foot type, biomechanical scenario, and desired functional performance expectations. PMID- 11417152 TI - Rehabilitation of plantar fasciitis. AB - Rehabilitation of plantar fasciitis can be a lengthy and sometimes difficult process. The patient and the practitioner can become discouraged by slow progress. It is of benefit to the patient and practitioner to be able to follow a logical approach in the rehabilitation plan. No one modality of treatment for this condition has been shown to be effective in all instances. It is best to choose several complementary treatment modalities in the rehabilitation of this difficult condition. PMID- 11417153 TI - Achilles' tendonitis. An overview and reconditioning model. AB - Achilles' tendon injuries are common in runners. The muscle tendon complex primarily functions eccentrically, and it is during this phase that failure occurs. Reconditioning programs that incorporate eccentric strengthening and plyometric and other neuromuscular strengthening theories with a concerted specific gradual activity challenge can be successful. To optimize return to activity, more work needs to be done to identify the amount of stress (i.e., exercise) that can be tolerated at a specific phase of the inflammatory process. PMID- 11417154 TI - Surgery of the Achilles' tendon. AB - Without question, injury to the Achilles' tendon is one of the commonest running injuries involving the lower extremity. Repair of acute and delayed ruptures is discussed. Tendonitis is by far commoner than ruptures. The pathology and conservative and surgical treatments are discussed. PMID- 11417155 TI - Stress fractures of the foot and ankle. AB - Stress fractures of the foot and ankle are common injuries that require a high index of suspicion to make the appropriate diagnosis. If required, advanced imaging modalities should be used to establish the diagnosis. When developing a treatment plan for the runner, the sports medicine physician must keep the athlete informed as to the nature of the injury and the necessity of the treatment. A fitness plan must be developed with the runner to ensure that strength, flexibility, and cardiovascular conditioning are maintained during this period. Ideally the physician should develop this plan with the runner and not dictate the type of activity that is used to obtain these goals. PMID- 11417156 TI - Diagnostic approach to chronic exercise-induced leg pain. A review. AB - The patient presenting with complaints of chronic exercise-induced lower leg pain can be a diagnostic challenge. Although myriad specialized diagnostic studies and modalities are at one's disposal, emphasis on a logical history and physical usually can identify the underlying cause without exposing the patient to any unnecessary testing. A well-directed history and physical, coupled with an understanding of the current state of knowledge regarding the pathophysiology and pathomechanics of shin splints, helps meet this challenge effectively. PMID- 11417157 TI - Running injuries about the knee. AB - Evaluation and treatment recommendations for a painful knee in a runner can be accomplished with a good knowledge of knee pathology, careful physical exam, and appropriate use of imaging tests. Most knee problems in a runner fall into one of the entities listed above. Fortunately, almost all can expect significant improvement with appropriate treatment. PMID- 11417158 TI - Pushing the envelope. Case studies on how fast you can and cannot return the elite athlete to running. AB - When treating an elite athlete, a physician always must keep in mind the status of the athlete in training, upcoming sports events, and the athlete's financial status. If the treatment requires the athlete to withdraw from regular training or sports events, a modified training program should be considered. This modified training must be balanced with returning the athlete back to full form and keeping him or her physically fit. PMID- 11417159 TI - Massage therapy for sprinters and runners. AB - Winning the gold medal is everything. The competitive sprinter or runner successfully navigates the road to the gold by relying on talent, dedication, and hard work. Along this steep and difficult road, massage therapy provides a powerful tool that helps the athlete reduce recovery time after a track meet, minimize injury, and improve body movement fluidity. PMID- 11417160 TI - Nutrition for runners. AB - Good nutrition plays an important role in a runner's overall health and fitness. The recommended diet for runners includes a proper balance of protein, carbohydrates, and fats to ensure good energy and performance. Specific requirements for vitamins and minerals are essential for recovery and preventing injuries. Fluids and proper hydration are key components in preventing dehydration, fatigue, and poor performance for all runners. PMID- 11417161 TI - Chiropractic approach to running injuries. AB - The chiropractic approach to the treatment of running injuries involves a thorough understanding of the anatomy, biomechanics, motor patterns, and kinetic chains of the lower limb, including the pelvis. By identifying the underlying dysfunction, the chiropractor can implement a multilevel treatment protocol that involves manipulative therapy, restoration of faulty biomechanics, strengthening of weakened muscle groups, and motor pattern re-education. PMID- 11417162 TI - Use of the lapidus bunionectomy in first ray insufficiency. AB - Surgical management of first ray insufficiency in HAV or hallux limitus is crucial to long-term success. Although technically demanding, the Lapidus fusion serves to address the deformity at its apex and restores normal weight bearing to the foot. This arthrodesis offers predictable, durable results in addressing various first ray deformities and occupies an important place in the foot and ankle surgeon's armamentarium. The authors' technique is described herein for the interest of and evaluation by surgeons performing this procedure. As clinicians recognize the importance of addressing proximal components of the HAV deformity, the Lapidus and other procedures will likely see increased use in the surgical management of first ray deformities. PMID- 11417163 TI - Eliminating nursing education departments revisited. PMID- 11417164 TI - Effect of nurses' experience and education on quality of care. PMID- 11417165 TI - Online nursing documentation: finding a middle ground. PMID- 11417166 TI - Soul + spirit + resources + leadership = results. PMID- 11417167 TI - Professional responsibility versus mandatory overtime. PMID- 11417168 TI - Developing innovative care models: the use of customer satisfaction scores. AB - With the increased emphasis on accountability, cost, and quality in healthcare, models of care delivery are being restructured. The author examines the planning, implementation, and evaluation of a model of care delivery for neonates based on customer, staff nurse, nurse practitioner, and attending physician perceptions of care and their suggestions for improvement. PMID- 11417169 TI - Critical access hospitals: rural nursing issues. AB - Critical Access Hospitals (CAHs) are a recent federal initiative to address the fiscal concerns of small rural hospitals and improve access to healthcare for rural residents. A national effort exists to examine the outcomes of this federal initiative, but there is a paucity of information about nursing in CAHs. This pilot study, using survey techniques, examined rural nurses' perceptions of CAH conversion. The authors discuss the authorizing legislation, Medicare Rural Hospital Flexibility Program (MRHFP), and highlight survey findings on nurses' perceptions about hospitals that converted to CAH status. The information can be used by nursing administrators and educators to prepare nurses to work in CAHs that are located in more remote areas of the United States. PMID- 11417170 TI - Time, task, and talents in ambulatory care nursing. AB - While the work of ambulatory nursing is increasingly complex, nurses have little specific preparation and support. Role confusion is prevalent, and nurses are often unable to articulate their roles. The authors present results of a survey that assessed practice patterns of registered nurses (RNs) in ambulatory care settings. They discuss how ambulatory RNs' perceptions that what they actually do in daily practice is disconnected from what they feel is important to do. Barriers to practice change are highlighted with recommendations that include working for effective practice design, building consensus about the scope of ambulatory RN practice, increasing education in academic and service settings, and enhancing support for delegation. PMID- 11417171 TI - Patient education: designing a state-of-the-art consumer health information library. AB - Many patients believe that the education they receive about their health and their illnesses is inadequate or lacking. Nurse executives are in a key position to influence their patients' abilities to become more informed and to take greater responsibility for their healthcare decisions. In the article, the authors discuss Massachusetts General Hospital's state-of-the-art consumer health information library, including how the project was planned, organized, and implemented. PMID- 11417172 TI - Hospital RNs' job satisfactions and dissatisfactions. AB - In a survey addressing work-related stress, 1,780 registered nurses (RNs) discussed the many variables affecting their job satisfaction, including: why they chose nursing, patient care issues, nurse manager roles, salaries and benefits, and intent to stay in nursing. Their responses may be painful for nursing management to read. Yet, if new candidates are to be attracted and existing RNs retained, satisfaction of RNs with their jobs must be considered. PMID- 11417173 TI - [Evaluation of clinical effectiveness of intravenous laser irradiation of blood, plasmapheresis and their combination in patients with bronchial asthma]. AB - AIM: To compare clinical response to intravenous laser radiation of blood (ILRB), plasmapheresis (PA) and ILRB + PA in patients with bronchial asthma (BA). MATERIALS AND METHODS: 122 patients with endogenic BA of moderate severity were divided into four groups: group 1 was exposed to ILRB, group 2--to PA, group 3- to ILRB + PA, group 4 received only chemotherapy. The effect was assessed by body plethismography, peak flowmetry, NBT-test, LCP-test. RESULTS: Patients of group 1 3 vs group 4 demonstrated earlier disappearance of cough and normalization of lung auscultative picture, lower demand of oral glucocorticosteroids, 2 times longer remission, better external respiration function, earlier normalization of peak expiration flow. ILRB, PA and ILRB + PA activate function of oxygen dependent bactericidal system of blood neutrophils and inhibited activity of non oxygen-dependent system. CONCLUSION: Combined treatment of moderate severity BA with ILRB, PA alone and in combination is more effective than conventional drug therapy, the highest effect by remission terms being achieved in combination of ILRB with PA. PMID- 11417174 TI - [Clinical effectiveness of superoxide inhalations and their effects on crystalline structure and antiradical defense of blood serum and expired air condensate in patients with bronchial asthma]. AB - AIM: To raise efficiency of combined treatment of bronchial asthma (BA) by application of intranasal inhalations of gaseous superoxide (GSO) basing on clinicolaboratory, functional and biochemical indices. MATERIAL AND METHODS: The study entered 20 healthy subjects and 57 BA patients. All of the patients received conventional treatment, but 30 of them additionally inhaled GSO. The effect of inhalations was assessed by parameters of LPO products, enzymatic and nonenzymatic antiradical defense (ARD) in the blood and expired air condensate (EAC), morphometry of EAC crystalline structures. RESULTS: GSO intranasal inhalations improve treatment of BA as it produces positive effect on endogenic oxidative stress, LPO, enzymatic and nonenzymatic ARD, blood and EAC tesigraphy. CONCLUSION: Antiinflammatory effect of GSO in BA is suggested. PMID- 11417175 TI - [Changes in biochemical inflammation markers in evaluation of the effectiveness of basic chemotherapy in bronchial asthma]. AB - AIM: To analyse informative value of monitoring of NO metabolites concentration in condensate of expired air vapor (EAVC) for definition of bronchial asthma (BA) severity and antiinflammatory effect of basic therapy. MATERIAL AND METHODS: NO metabolites concentration in EAVC was measured with Grace's reagent in 76 adults and 180 children in the course of basic therapy with flixotide, tailed, ketotiphen with intal. RESULTS: NO metabolites occurred in high concentrations in EAVC of both children and adults. These concentrations were the highest in attacks and in severe BA and were reduced by basic therapy. This effect differed with the drug and its dose, e.g. flixotide and tailed given to children for a month reduced NO metabolites close to concentrations observed in healthy subjects. Ketotiphen and intal for 6 months failed to low NO metabolites significantly. CONCLUSION: NO secretion monitoring is sensitive in assessment of respiratory inflammation in BA and is informative in assessment of effectiveness of the basic therapy. In moderate BA children on flixotide and tailed were the first to achieve normal NO secretion in the airways. PMID- 11417176 TI - [Chronic obstructive lung diseases in patients with ischemic heart disease: a 15 year study]. AB - AIM: To characterize acquired chronic obstructive pulmonary diseases (COPD) in patients with ischemic heart disease (IHD). MATERIAL AND METHODS: Monitoring of external respiration function (ERF) was performed for 15 years in 1552 IHD patients on combined drug treatment. RESULTS: The 15-year follow-up has revealed that IHD patients treated with beta-blockers develop pathological changes in respiration pattern manifesting primarily with dyspnea, obstructive and mixed ventilatory disorders, syndrome of early expiratory obstruction in affected small airways due to subnormal lung elasticity, defective diffuse function, signs of terminal respiratory failure. CONCLUSION: IHD patients taking drugs need monitoring of ERF. Optimal treatment consists in early diagnosis of IHD and COPD, adequate combined therapy with beta-blockers and bronchodilating drugs. PMID- 11417177 TI - [Prognostication of obstructive syndrome in patients with chronic bronchitis considering hereditary factors]. AB - AIM: To propose method of accurate prediction of chronic obstructive pulmonary disease (COPD) at early stages of development. MATERIAL AND METHODS: 92 males with COPD and 55 males with chronic non-obstructive bronchitis aged 38 to 72 years were examined according to a technique which identifies hereditary predisposition to multifactor diseases and using an algorithm based on multivariate analysis. RESULTS: An original mathematical model is proposed which can quantitatively examine and range by influence the role of environmental and hereditary factors in development of COPD. CONCLUSION: Detection of hereditary predisposition to COPD may be one of the objective criteria to recommend giving up smoking and for occupational orientation. PMID- 11417178 TI - [Correlations between electrophysiological parameters of the heart, blood gas composition and hemodynamics in patients with chronic obstructive bronchitis]. AB - AIM: To elucidate correlations between gas composition of blood, hemodynamic and electrophysiological parameters of the heart and effect of these indices on arrhythmia onset in patients with chronic obstructive bronchitis (COB). MATERIAL AND METHODS: "Rhythm" ECG, 24-h ECG monitoring, transesophageal pacing, heart ultrasonography were performed in 160, 67, 160 of 160 patients with chronic obstructive bronchitis (COB), respectively. 76 patients suffered from respiratory insufficiency (RI) degree 1 and 84 of RI degree 2. 58 patients free of RI served control. Gas composition of the blood and acid-base balance of the capillary blood were examined in 50 patients. RESULTS: Shortening of effective refractory atrial period and prolongation of interatrial conduction predispose to emergence of suptaventricular arrhythmia in COB patients with RI. With the onset and development of RI in COB patients, heart chamber size and myocardial function become more and more essential for cardiac electrophysiological parameters. For COB patients free of RI, gas composition of capillary blood is of key importance in this respect. CONCLUSION: The proposed complex clinicofunctional method for assessment of electrophysiological condition of the heart with evaluation of intracardiac hemodynamics and gas composition of the capillary blood is recommended for registration of signs of the onset and unfavorable progress of cardiac arrhythmia in COB patients with RI as well as for objective and safe control over effectiveness of treatment of such patients. PMID- 11417179 TI - [Effects of bacterial extract IRS-19 on the concentration of hydrogen peroxide and myeloperoxidase activity in nasal washings of patients with chronic bronchitis]. AB - AIM: To determine the effect of intranasal treatment with IRS-19, an immunomodulating agent, on the number of polymorphonuclear leukocytes (PMNL), H2O2 concentration and myeloperoxidase (MPO) activity in nasal washings. MATERIAL AND METHODS: 28 adult patients of both sexes with chronic bronchitis participated in an open study of intranasal treatment with IRS-19. RESULTS: The number of PMNL recovered from nasal spaces increased from 4460 +/- 3960 to 10,490 +/- 10,950 cells/ml (p < 0.02) after two month administration of IRS-19. It was accompanied by 2.6- and 1.4-fold increase (p < 0.001) in MPO activity and H2O2 concentration, respectively. However, no correlation was found between increments in these three variables. CONCLUSION: Since PMNL and MPO--H2O2--Cl- system are involved in the first line of defense against invading pathogens it is suggested that the above mentioned changes may represent one among mechanisms leading to enhancement of antibacterial defence in the airways in response to treatment with IRS-19. PMID- 11417180 TI - [Characteristics of expression of active oxygen forms by blood cells in patients with chronic bronchitis]. AB - AIM: To study generation of active oxygen forms in the whole blood of patients with chronic bronchitis and correction of the detected disorders. MATERIAL AND METHODS: By means of registration of spontaneous and staphylococcus-induced luminol-dependent chemiluminescence (LDCL) of the whole blood, generation of active oxygen forms in 73 patients with chronic bronchitis (CB) has been studied. RESULTS: By the intensity of luminescence, all the examinees were divided into two groups. In group 1 LDCL was high, in group 2 LDCL was under the control level. Combined treatment of group 1 patients included inhalations of ceruloplasmin antioxidant. Patients of group 2 were administered sodium hypochlorite that possesses prooxidation properties. Compared to conventional treatment, the above two treatments produced marked positive changes in the studied indices and clinical symptoms. CONCLUSION: The method of chemiluminescence registration of the whole blood allows validation of pathogenetic therapy of patients with CB and improves outcomes of the treatment. PMID- 11417181 TI - [Effectiveness of lasolvan in patients with chronic obstructive bronchitis and non-atopic bronchial asthma]. AB - AIM: To define efficiency and mechanisms of action of mucoregulator lasolvan. MATERIAL AND METHODS: Lasolvan treatment was given to 50 patients with chronic nonspecific pulmonary diseases (CNPD). External respiration function (ERF) and viscosity of sputum were examined on Bodyscreen II and viscosimeter Reotest 2, respectively. RESULTS: Maximum effect was achieved on day 2-3 of lasolvan treatment: resistance of airways decreased 1.5-fold, forced pulmonary capacity and peak expiration flow increased. ERF improvement was associated with subjective improvement. Mean sputum viscosity remained unchanged. Reduction of minimal viscosity was found after the first intake of lasolvan. Later, maximal viscosity was higher in most of the patients. By day 7-10 amount of the sputum discharged forn 24 hours rose while maximal viscosity fell. CONCLUSION: Lasolvan is an effective mucoregulator which perfectly improves rheologic properties of bronchial secretion. Its ability to eliminate pathological bronchial secretion from small bronchi is of special importance. This allows to use lasolvan for prevention of cor pulmonale in CNPD patients. PMID- 11417182 TI - [Severe forms of bronchial asthma]. PMID- 11417183 TI - [Obstructive sleep apnea in patients with stage I arterial hypertension]. AB - AIM: To study sleep breathing in young men with arterial hypertension stage I. MATERIAL AND METHODS: Somnological history, night cardiorespiratory monitoring, 24-h monitoring of arterial pressure (AP) and ECG were analysed in 34 young men with arterial hypertension (AH) of the first degree and 12 normotensive young men. RESULTS: Hypertensive young men had a higher index apnea/hypopnea, mean duration of apnea/hypopnea (p < 0.05), maximal desaturation (p < 0.1), indirect parameters of inspiratory apnea, they snored more and woke up in sleep more frequently, their night hypertension was more than daytime. This was observed even in patients with overweight. CONCLUSION: All hypertensive young men examined had indirect signs of obstructive sleep apnea. The triggering role in it of the lungs and subclinical obstruction of upper airways in AH is suggested. PMID- 11417184 TI - [Clinical polymorphism in respiratory oxalosis]. AB - AIM: Examination of clinical polymorphism of chronic obstructive pulmonary diseases (COPD) in defects of oxalate metabolism to make diagnostic outpatient screening of the preclinical stage. MATERIAL AND METHODS: Diagnostic dysgenetic markers of respiratory oxalosis (RO)--red hair in monthers and 24-h oxaluria- were studied in 28 women and 7 men. 8 women (group 1) had diagnostic association, 7 women (group 2) had no hereditary marker, 13 women (group 3) had no signs of disturbed oxalate metabolism. In addition, families of group 1 patients were examined for preclinical signs of visceral oxalosis in close relatives (kinship degree I). A comparison was made of quantitative enzyme assay of registering 24-h oxaluria (Lartillot M. et Vogel G) and titration by G. A. Sivorinovsky. RESULTS: Group 1 COPD patients with mild disease had rather high 24-h oxaluria. In group 2 and 3 patients oxaluria was significantly lower. Dysgenetic markers--24-h oxaluria with the hereditary criterium--may be used in differential diagnosis of RO with its phenocopy having a more severe course at preclinical stage. Male relatives of kinship degree I had significant differences with group 1 patients in 24-h oxaluria, oxaluria was combined with clinical symptoms of acid, uratic diathesis. CONCLUSION: The enzyme assay of oxalate in 24-h urine in combination with hereditary marker is an adequate screening method for preclinical stage of RO. The presence of various clinical manifestations of visceral oxalosis--RO and acid, uratic diathesis in the family--may indicate clinical polymorphism of mutant gene. PMID- 11417185 TI - [Clonazepam in the treatment of labile arterial hypertension in the elderly]. AB - AIM: To investigate effectiveness of clonazepam and its influence on autonomic function. MATERIAL AND METHODS: 56 patients with LHE aged 67.0 + 6.3 years were examined. 50 of them were treated with clonazepam in daily dose 1-2 mg. Their results were compared with age and sex matched normotensive persons. The routine clinical examination, regular measurement of blood pressure (BP), psychological tests, investigation of heart rate variability and evoked cutaneous sympathetic potentials (ECSP) were made. These tests were performed before and after treatment with clonazepam. RESULTS: Clonazepam significantly reduced fluctuations of BP in 82.0% of patients with LHE: the fluctuation range of systolic BP was reduced 2 times, diastolic BP--1.6. At the same time clonazepam diminished the level of anxiety, heart rate variability with increasing normalized VLF (central influence) and lowering LF and HF (segmental sympathetic and parasympathetic influences). ECSP were characterized by slowing sympathetic conduction and reduction of ECSP amplitude. CONCLUSION: Clonazepam, by changing autonomic properties, leads to stabilization of blood pressure in most patients with LHE. It is recommended to use clonazepam in daily dose 1-2 mg in combination with conventional antihypertensive medications for prevention of excessive fluctuation of BP in patients with LHE. PMID- 11417186 TI - [Primary antiphospholipid syndrome and severe pulmonary hypertension without thrombosis of arteries of the lesser circulation]. PMID- 11417187 TI - [Use of glucocorticosteroids in asthmatic conditions (review)]. PMID- 11417188 TI - [Empirical antibacterial therapy of nonhospital pneumonia (review of foreign literature)]. PMID- 11417189 TI - [Methods of studying the mucociliary system: possibilities and perspectives]. PMID- 11417190 TI - [Clinical course, internal picture of the disease and pathogenic reactions in patients with bronchial asthma]. AB - AIM: To study clinical features of bronchial asthma (BA) associated with internal picture and nosogenic reactions. MATERIAL AND METHODS: Clinical features of BA were studied in 108 patients (38 males and 70 females, mean age 44.7 +/- 1.4 years, mean BA duration 6.6 +/- 0.76 years) treated in the clinic of the I. M. Sechenov Moscow Medical Academy in 1995-1998. RESULTS: Clinical and statistical analysis of BA clinical course allowed to single out three leading variants of BA course: persistently obstructive (stable and definite respiratory disturbances with progressive deterioration of bronchial permeability), moist (associated chronic bronchitis, allergic rhinosinusopathy, chronic maxillary sinusitis, intestinal dysbacteriosis, repulsive symptoms, e.g. discharge of much sputum et cet), paroxysmal (short-term episodes of asphyxia arrested by inhalations of beta 2-agonists, absence of stable respiratory disorders). The above variants of BA course significantly correlated with types of the internal picture (IP) and types of nosogenies. Persistently obstructive, moist and paroxysmal BA variants were characterized with vital, defensive and coping IP, respectively, neurotic reactions with hypochondria, hypochondriac depressions; pathocharacterological sensitive reactions; neurotic and affective reactions with "la belle indifference" and "euphoric pseudodementia", respectively. CONCLUSION: The above correlations can be used for planning and conduction of psychocorrective measures aimed at optimisation of patient-doctor compliance. PMID- 11417191 TI - [The epidermal permeability barrier and its function in immune reactions]. AB - Epidermal barrier represents one of the essential conditions of life on Earth. However, our present understanding of its function is still insufficient. The role of water incorporated in stratum corneum has not been yet studied enough. Till recently, both the stratum corneum et granulosum were considered to be "dead" layers. It turned out that these layers form very active part of epidermis and they have a secretory function. Their rigid anhydrotic consistency, complex intercellular spaces, and lipid bilayers produced by lamellar bodies form the defense barrier against both the loss of fluids and ions and protecting against the hostile external environment. Important stabilizing function in the barrier has ceramides and urea. The Langerhans' dendritic cells and macrophages induce production of cytokines in keratinocytes by means of calcium signals. That triggers the inflammatory cascade, which is accompanied by formation of pathological types of T-cells and by activation of apoptosis. Last but not least, the impairment of epidermal barrier has an important role in the physiological aging of the skin. PMID- 11417192 TI - [Antioxidant effects of melatonin]. AB - Increase in knowledge about reactive oxygen species action mechanisms and oxidative stress effects in living organisms led to intensive seeking for new, more effective substances, which prevent extreme development of oxidative stress or are able to decrease its negative influence, damaging cell structures and many cell functions. These substances are called antioxidants, scavengers, trappers or quenchers. In nineties, melatonin became the centre of the interest in the filed of investigation of antioxidative properties of different chemical substances. This is in living organisms ubiquitous substance with relatively simple chemical structure, good physical properties and wide physiological effects. The main role of endogenous melatonin comprises receptor-mediated biological rhythms synchronisation. Among other functions mentioned later belong anti-gonadotropic, immunotropic and non-receptor-mediated antioxidative effects. Melatonin is said to have also antineoplastic properties. Its anti-aging effect is discutable. PMID- 11417193 TI - [Effect of angiotensin-converting enzyme insertion-deletion polymorphism on progression of renal and cardiovascular diseases]. AB - Many controversial studies concerning relation between angiotensin converting enzyme polymorphism and renal and cardiovascular disease have been published during the last years. Most of the papers have suggested that the DD genotype plays an important negative role in the progression of some renal diseases (e.g. IgA nephropathy, diabetic nepropathy). The D allele may be an independent risk factor for development of the target organ damage in essential hypertension. The therapeutic response on inhibitors of angiotensin converting enzyme depends on insertion-deletion polymorphism. It probably also depends on the gender. The pathological mechanisms of insertion-deletion polymorphism have not yet been clearly identified. PMID- 11417194 TI - [Nitric oxide in patients after cadaveric kidney transplantation]. AB - BACKGROUND: The role of nitric oxide (NO) after cadaveric renal graft transplantation has not been yet fully clarified. The aim of our study was to examine NO production into the urine of patients following cadaveric renal graft transplantation with a normal course and complications (acute rejection and cyclosporin toxicity). METHODS AND RESULTS: Production of stabile NO metabolites (NO2 and NO3) into urine (U-NOx) was examined in recipients of cadaveric renal transplantation. Only patients with standard triple immunosuppressive therapy (cyclosporin, azathioprine, prednisone) were include into the study. Patients receiving other immunosuppressive agents or drugs affecting NO formation (nitrates, ACE inhibitors) were excluded from the study, as were those with infectious or other serious post-transplant complications. Overall, we examined 33 patients (21 men and 12 women), with acute rejection and cyclosporin-induced toxicity in ten each, and a normal course with no complications in 13. The mean age of the patients was 50.96_11.13 years. U-NOx was examined by biochemistry using Griesse reaction every day after transplantation both in a morning urine sample and in a sample from 24-hour collection over the preceding day and calculated to 1 mmol/l of urinary creatinine (U-Cr). The levels of U-NOx/U-Cr in patients with acute rejection over the past 2 days before its development were lower compared with those in patients with a normal course (p_0.05). No difference was found between the groups of patients with cyclosporin-induced toxicity and a normal course. The levels of U-NOx were inversely correlated (p_0.01) to the levels of serum creatinine (S-Cr), but did not correlate with the blood levels of cyclosporin A. CONCLUSIONS: The study demonstrated a decrease in urinary U-NOx production within the past 2 days before renal transplant rejection. The levels of U-NOx in patients with cyclosporin-induced toxicity remain unaltered. U-NOx/U-Cr could possibly become a non-invasive marker of rejection. PMID- 11417195 TI - [Coital behavior in men with varicocele]. AB - BACKGROUND: According to the concept of Partial Androgen Deficiency of the Aging Male (PADAM), testosterone supplementation becomes recommended in prevention and therapy of different disturbances especially in aging males. METHODS AND RESULTS: In the group of 58 married patients with varicocele and in 85 married normozoospermic men the plasma testosterone levels were estimated and the coital activity was determined by an interview. Plasma values of male sex hormone of patients with varicocele were significantly lower (p < 0.01) than those of controls. The coital activity of both groups was not significantly different. CONCLUSIONS: The hypothesis is presented expressing that Nature ensuring the existence of the human species produces in an over-supply not only of spermatozoids, but also of the male sexual hormones. PMID- 11417197 TI - [Compression syndromes]. PMID- 11417196 TI - [Founding of the Society of Czechoslovak Physicians in Bratislava in 1920- contribution of Czech physicians in the mutual development and cooperation in Czech and Slovak medicine]. AB - Foundation of the Society of the Slovak Medical Doctors (SSMD) in Bratislava in June 13, 1920 (till 1938 Society Czechoslovak Medical Doctors) belongs to the most important milestones in the modern history of the Slovak medicine and it can be directly related to the foundation of Comenius University and its first faculty--Faculty of Medicine. SSMD started in October 1921 publishing the Bratislava Medical Journal, which represented the first scientific medical journal in Slovakia after the foundation of the Czechoslovak Republic. Eighty years of existence of SSMD, which belongs with its almost 2500 members to the largest professional society within the Slovak Medical Society, testifies its contribution to the development and growth of the Slovak medicine. The Society had been for many years the only professional forum where the Slovak medical doctors could meet and where the scientists of the Medical Faculty in Bratislava professionally matured. It represents also an important evidence of the material and spiritual assistance and collaboration of the Czech and Slovak medical doctors during the First Czechoslovak Republic. PMID- 11417198 TI - Neural network in communication with medical computer system. AB - The paper presents a concept of an experimental module designed to recognize spoken utterances that cover a limited range of words indispensable in dialogs with computer medical systems. Research into the recognition of spoken words by a module based on artificial neural network is described. Usefulness of the obtained results for surgery-assisting multimedia systems and for a patient simulator supporting medical education of students in case history-taking and diagnosing is also discussed. PMID- 11417199 TI - A neural network approach in diabetes management by insulin administration. AB - Diabetes management by insulin administration is based on medical experts' experience, intuition, and expertise. As there is very little information in medical literature concerning practical aspects of this issue, medical experts adopt their own rules for insulin regimen specification and dose adjustment. This paper investigates the application of a neural network approach for the development of a prototype system for knowledge classification in this domain. The system will further facilitate decision making for diabetic patient management by insulin administration. In particular, a generating algorithm for learning arbitrary classification is employed. The factors participating in the decision making were among other diabetes type, patient age, current treatment, glucose profile, physical activity, food intake, and desirable blood glucose control. The resulting system was trained with 100 cases and tested on 100 patient cases. The system proved to be applicable to this particular problem, classifying correctly 92% of the testing cases. PMID- 11417200 TI - Evaluating training data suitability for decision tree induction. AB - Decision tree induction, as well as other inductive learning methods, requires training data of high quality to be able to generate accurate and reliable classification models. Example cases should form a representative sample from the application area, and the attributes used to describe example cases should be relevant and adequate for the classification task to be solved. In this paper, measures of the strength of association and an entropy-based approach have been used to assess the quality of the training data. Studied classification tasks related to three otological data sets: a conscript data set, a vertigo data set, and a postoperative nausea and vomiting data set. The paper suggests that the studied approaches give some guidelines about the quality of the training data, but other approaches are also needed to guide training data building. PMID- 11417201 TI - Database reusability in intelligent medical systems. AB - Reuse or reusability is not a specific, algorithmic, heuristic or only a simple set of guidelines. Database reuse means the use of an existing component--a database entity--in a new context, either elsewhere in the same system or in another system. According to different definitions, an intelligent system is a "power tool for thinking"; but on the other side it is only a kind of information system with built-in knowledge to support decisions made by human experts. Similar conclusions could be made for intelligent medical systems and introduce the database reusability in this environment with a purpose to increase the quality of an intelligent medical system. In the paper the problem of the database reusability will be presented more in detail, especially its integration in an intelligent medical system. Finally, the results of such integration and the benefits for the medicine will be discussed. PMID- 11417202 TI - Issues in the design of medical ontologies used for knowledge sharing. AB - Recent work in Medical Informatics is exploring the development and the use of formal ontologies as a way of specifying content-specific agreements for the sharing and reuse of knowledge among several computer systems. We describe the role of ontologies in supporting knowledge sharing activities in medicine Principles for the design of ontologies have been proposed, mainly in other domains: these principles include parsimony, clarity, representation of categories versus terms, and coherence. We analyze how and why these principles can or cannot be applied from case studies from medical systems. Regarding the fact that most of medical concepts are empirical, selected design decisions are discussed. An alternative representation choice consists in mapping principled general core ontologies and domain ontologies. PMID- 11417203 TI - Orthodontic tooth movement in the mixed dentition. Histological study of a human specimen. AB - MATERIAL AND METHOD: We present a retrospective analysis of periodontal tissue reactions and clinically relevant phases of permanent tooth eruption and deciduous tooth resorption after half a year of orthodontic tooth movement in the upper and lower jaw of a deceased male (age 9 years 3 months). Specimens of the horizontal plane (lower jaw) and sagittal plane (upper jaw) were prepared by the ground microsection technique without prior decalcification. RESULTS: Histologically, reactions in the periodontal ligament presented as characteristic appositional/resorptive metaplastic processes in the areas of tension and pressure, with side effects such as root resorption and periodontal necrosis being only minor. In the upper jaw, an erupting permanent canine was closely associated with the tooth germ of the first premolar. Resorptive follicle activity had resulted in extensive erosion of the interradicular bone and root resorption in the area of the first premolars. CONCLUSION: These findings suggest that it might prove useful to take critical topographic findings as parameters for differential therapeutic decisions. Follicle-driven deciduous tooth resorption presenting partly as resorption lacunae and partly with linear characteristics was accompanied by resorptive/appositional remodeling of the alveolar socket as well as regressive changes in the supracrestal fibers of the marginal gingiva. PMID- 11417204 TI - Bone remodeling of the human mandible during prenatal development. AB - MATERIAL AND METHOD: Based on histological serial sections of human fetuses (22 117 mm crown-rump length) the remodeling of the bone surfaces of the fetal mandible was characterized and visualized in 3D reconstructions. RESULTS: In the embryo of 22 mm crown-rump length the mandible was completely covered by cells producing bone apposition, but beginning with the embryo of 25 mm crown-rump length, morphological differentiations of these cells leading to bone modeling were visible. These modeling processes were found to be much more complicated than described in the literature so far. CONCLUSION: Bone growth of the fetal mandible is a complex process comprising not only lingual resorption and buccal apposition. Instead, we found well differentiated remodeling processes, represented by variable portions of apposition, resorption and resting areas seamed by lining cells. PMID- 11417205 TI - The influence of cephalometric parameters on resonance of speech in cleft lip and palate patients. An interdisciplinary study. AB - There are only few studies concerning the correlation between craniofacial morphology and resonance of speech in cleft palate patients. Moreover, these investigations show a considerable inhomogeneity in material and method, their basic approach was predominantly retrospective, and the statistical methods were restricted to univariate procedures. Thus, the aim of the present study was twofold: firstly to clarify the extent to which correlations exist between craniofacial morphology and resonance of speech in cleft palate patients, subject to a sufficiently large number of patients being available to ensure differentiation with regard to age and cleft type, and secondly, within a prospective longitudinal study in juvenile cleft palate patients, to investigate whether pubertal craniofacial growth changes result in changes of resonance. The collective comprised 137 cleft palate patients. The following investigations were performed: lateral cephalometry, nasometry, and standardized speech recording. Additionally, these investigations were repeated in 51 juvenile patients after a minimum time interval of 2 years. In order to analyze the complex relations between craniofacial morphology and degree of hypernasality, multivariate statistical procedures were applied. The results of the present study indicate complex correlations between cephalometric parameters and resonance of speech, requiring age-specific differentiation. In this regard, the ratios between the length of the soft palate and the sagittal depth of the nasopharyngeal airway were of prime importance. Beyond this, significant correlations were found between craniofacial growth changes and changes of resonance during puberty which might be influenced both by dentofacial orthopedics and by maxillofacial surgery. PMID- 11417206 TI - Comparison of modified Teuscher and van Beek functional appliance therapies in high-angle cases. AB - The aim of this study was to investigate the effects of modified Teuscher and van Beek functional appliances on the skeletal and dento-alveolar pattern in high angle Class II, Division 1 patients. The collective consisted of 32 patients with a high-angle Class II, Division 1 malocclusion. The modified Teuscher activator was applied to a group of twelve patients, and the van Beek activator to a group of ten patients. A control group of ten subjects was used for comparison purposes. The mean chronological ages of the groups were 12, 11.8 and 11.5 years, respectively. 64 cephalograms taken before and after the treatment were evaluated. Intra-group measurement relations were determined by the Wilcoxon test, and inter-group relations by analysis of variance and Duncan tests, using SPSS statistical software. The following results were observed for both treated groups without any change in growth direction: inhibition of maxillary growth, stimulation of mandibular growth, retrusion of upper incisors, distal tipping of upper molars, and reduction of overjet and overbite compared with the control group. The decrease in overbite in the van Beek group was due mainly to intrusion of the incisors, and in the modified Teuscher group to molar extrusion. Both activators were considered preferable in terms of vertical control of the facial height in high-angle cases with deep overbite. However, stimulation of mandibular growth in the modified Teuscher activator group was found to be more significant than in the van Beek activator group. PMID- 11417207 TI - Self-etching primers--an alternative to the conventional acid etch technique? AB - MATERIAL AND METHODS: The aim of the present in vitro study was to evaluate the bonding capacity of three self-etching primers (Clearfil SE Bond [SE], Clearfil Liner Bond 2V [CLB], Novabond [NOVA]) to enamel. Two adhesive systems with a separate phosphoric acid etching procedure (Gluma Comfort Bond [GLU], Kurasper F [KU]) served as controls. 60 human incisors were used to evaluate shear bond strengths of composite cylinders to enamel. RESULTS: Significant differences were found between the groups (p < 0.001). The mean value for groups GLU, KU, CLB and SE was 24 MPa. The shear bond strength recorded for the Novabond system was significantly lower (17 MPa). CONCLUSION: The results of the present investigation indicate that enamel bonding especially with the self-etching primers CLB and SE is as effective as conventional phosphoric acid etching. PMID- 11417208 TI - Optimizing interdisciplinary cooperation for patients with orofacial dysfunctions. Presentation of an interdisciplinary diagnostic referral sheet. AB - BACKGROUND: The quality of interdisciplinary cooperation is one of the most important factors in the success and long-term stability of treatment of patients suffering from orofacial dysfunctions. However, speech pathologists have criticized the poor communication and diagnostics at the time of referral and have requested a standardized diagnostic referral sheet with the aim of improving interdisciplinary cooperation. DIAGNOSTIC REFERRAL SHEET: The diagnostic referral sheet presented here is based on the demands of speech pathologists focusing on myofunctional therapy and helps in initial assessment of the individual situation and necessary treatment. It is subject to continuous updating and coordinates the many medical disciplines involved on account of the complex pathology. CONCLUSION: The multidisciplinary diagnostic referral sheet, which is independent of treatment methods and compressed into one page, is aimed at contributing to quality assurance and to the improved documentation of orofacial dysfunctions. PMID- 11417209 TI - Stretch-induced cell damage in sarcoglycan-deficient myotubes. AB - Sarcoglycans (SGs) are components of the dystrophin-glycoprotein complex, genetic defects in which cause skeletal muscle dystrophy and cardiomyopathy in humans and animals. To obtain insight into the roles of SGs, we characterized properties of myotubes prepared from cells of the rat L6 line or primary myoblast cultures of rat gastrocnemius muscle that were made SG-deficient by treatment with antisense oligodeoxynucleotides (AS-ODNs). Immunoblot and immunoprecipitation analyses revealed that dystrophin and its remaining associated proteins were tightly associated in these cells despite SG deficiency. 45Ca2+ influx into SG AS-ODN treated L6 myotubes under resting conditions was significantly higher (1.7-fold at 6 min) than in controls, suggesting that Ca2+ influx is activated in these SG deficient myotubes. When these cells were subjected to cyclic elongation of up to 20% for 1 h, a marked increase in creatine phosphokinase (CK) release into the medium was observed. Nifedipine, tranilast, FK506 and E64 or intracellular loading with 1,2-bis(2-aminophenoxy)ethane- N,N,N',N'-tetraacetic acid, tetrakis(acetoxymethyl)ester (BAPTA/AM) reduced the stretch-induced CK release; a raised extracellular [Ca2+] increased CK release. The stretch-induced damage to SG-deficient myotubes thus appears to be caused by alterations in cell Ca2+ homeostasis. A similar abnormality in Ca2+ handling has been reported for myoctes from mdx mice or dystrophin-deficient patients, in whom SGs are also greatly reduced or absent. Thus it is possible that SG deficiency may play a critical role in the pathology of dystrophin-deficient muscle. PMID- 11417210 TI - The effects of the inotropic agent EMD 57033 on activation and relaxation kinetics in frog skinned skeletal muscle. AB - The effect of the cardiotonic sensitizing drug EMD 57033 was studied in frog skinned skeletal muscle fibres at 12 degrees C to provide a baseline for skeletal muscle studies and for comparison with cardiac fibres. The activation and relaxation of fibres were induced by laser flash photolysis of the caged calcium NP-EGTA, and caged calcium chelator diazo-2 respectively. EMD 57033 (10 microM) slightly increased the rate of relaxation (rate constant k1 changing from 24.0 +/ 2.9 s-1 in control to 28.1 +/- 3.2 s-1) but had no significant effect on the rate of activation (k1 = 9.6 +/- 0.9 s-1 in control conditions, 9.7 +/- 1.6 s-1 with EMD 57033). The effect of the optical isomer of EMD 57033, EMD 57439, was examined on steady-state force and relaxation rate. EMD57439 (10 microM) slowed the rate of relaxation (k1 = 20.5 +/- 2.4 s-1) but had no effect on the maximal calcium-activated force whereas EMD 57033 increased it by 16.5 +/- 5.7%. These results are compared to earlier results from this laboratory in guinea-pig skinned trabeculae, and a possible model for the action of EMD 57033 whereby the drug enhances force per cross-bridge is discussed. PMID- 11417211 TI - Additive protective effects of late and early ischaemic preconditioning are mediated by the opening of KATP channels in vivo. AB - We investigated whether a combination of ischaemic late preconditioning (LPC) and ischaemic early preconditioning (EPC) induces additive myocardial protection in vivo, and the role of ATP-sensitive K (KATP) channels in ischaemic LPC and in LPC + EPC. Sixty rabbits were divided into seven groups. Anaesthetized animals were subjected to 30 min of coronary artery occlusion and 120 min of reperfusion (I/R). Controls (CON, n = 9) were not preconditioned. LPC (n = 10) was induced in conscious rabbits by a 5-min period of myocardial ischaemia 24 h before I/R. The KATP channel blocker 5-hydroxydecanoate (5-HD, 5 mg/kg) was given 10 min before I/R with (LPC + 5-HD, n = 9) or without LPC (5-HD, n = 8). EPC (n = 8) was induced by a 5-min period of myocardial ischaemia 10 min before I/R. Animals received LPC and EPC without (LPC + EPC, n = 8) or with 5-HD (LPC + EPC + 5-HD, n = 8). LPC reduced infarct size (IS, triphenyltetrazolium staining) from 57 +/- 11% (MW +/- SD, CON) of the area at risk to 31 +/- 19% (LPC, P = 0.004). 5-HD did not affect IS (5-HD: 60 +/- 12%, P = 0.002 versus LPC), but abolished the cardioprotective effects of LPC (LPC + 5-HD: 62 +/- 18%, P = 0.001 versus LPC). EPC reduced IS to 18 +/- 8%. Additional LPC led to a further reduction to 8 +/- 4% (LPC + EPC, n = 8; P = 0.005 versus EPC; P = 0.004 versus LPC). 5-HD abolished this additional cardioprotective effect of LPC + EPC (LPC + EPC + 5-HD, n = 8; 46 +/- 11%, P < or = 0.001 versus LPC + EPC). We conclude that the combination of ischaemic LPC and EPC induces additive cardioprotection. KATP channel opening mediates the cardioprotective effects of ischaemic LPC and LPC + EPC. PMID- 11417212 TI - Molecular cloning and expression of cERG, the ether a go-go-related gene from canine myocardium. AB - Given the anatomical and physiological similarities to the human heart, canine in vivo heart models may facilitate the analysis of molecular mechanisms underlying cardiac repolarization abnormalities. The development of such models depends, however, on information about canine K+ channels responsible for the establishment of IK currents. In this context, we isolated and sequenced the reverse transcript of the canine ether a go-go-related gene (cERG). The complementary deoxyribonucleic acid (cDNA-derived cERG polypeptide consists of 1,158 amino acids, the sequence of which shows striking homology to human, rat and mouse ERG subunits (97%, 94% and 95% identity respectively). In highly conserved peptide domains like the PAS domain, the membrane-spanning segments S1, S3-S6 and the pore-forming region, there was 100% identity. Analysis of cERG transcription revealed abundant expression of cERG messenger ribonucleic acid (mRNA) in heart and brain and low expression in liver, spleen and kidney. Membrane currents recorded in Xenopus oocytes expressing cERG channels showed functional properties very similar to the human K+ channel hERG, which encodes the alpha-subunit of the cardiac rapidly activating, delayed rectifier (IKr) channel. PMID- 11417213 TI - Single-channel properties of the sinoatrial node Na+ current in the newborn rabbit. AB - We have reported previously that the sinoatrial node (SAN) in the newborn rabbit expresses a Na+ current (INa) with properties similar to the neuronal type-I isoform and that this current contributes to the net inward current flowing during diastolic depolarization. To characterize this current further we conducted cell-attached single-channel experiments in isolated newborn SAN myocytes. The Na+ channel was sensitive to divalent cation block and had a single channel conductance of 25.6 pS in the absence of divalent cations. Kinetic compatibility between single-channel and previous whole-cell data was confirmed by measuring the time constant of current decay. At pacemaker potentials, time constants were of the order of tens of milliseconds. Additional experiments indicated that this slow inactivation arises because the Na+ channels expressed in the neonatal SAN tend to re-open frequently at potentials in the pacemaker range. We suggest that this is the mechanism by which a small tetrodotoxin (TTX) sensitive current contributes to the total inward current flowing during slow diastolic depolarization in neonatal (but not adult) pacemaker myocytes. PMID- 11417214 TI - L(+)-lactate does not affect twitch and tetanic responses in mechanically skinned mammalian muscle fibres. AB - This study investigated whether a high intracellular concentration of L(+) lactate (30 mM) affects normal excitation-contraction coupling in skeletal muscle. Electrical stimulation was used to elicit action potentials in the (sealed) transverse-tubular system of mechanically skinned muscle fibres, giving rise to twitch and tetanic force responses. As the sarcolemma was absent, lactate could be applied to the cytoplasmic environment via the bathing solution (at a constant pH of 7.1) and its effect examined independently of other metabolic changes that occur during muscle fatigue. The presence of 30 mM lactate had virtually no effect on direct activation of the contractile apparatus by Ca2+. Lactate also had no significant effect on either the rate of rise or the peak of the twitch response, with the only detectable effect being a slight (13%) slowing in its relaxation rate. As the amplitude of the twitch response (approximately 60% of maximum force) may be regarded as a sensitive indicator of the amount of Ca2+ released by an action potential, there was evidently to change in Ca2+ release in the presence of lactate. Lactate also had no significant effect on the rate of rise and peak force of the tetanic response or on its subsequent relaxation. Additional experiments, in which the sarcoplasmic reticulum was emptied of Ca2+ (in a caffeine solution) and reloaded repeatedly, showed no significant effect of 30 mM lactate on Ca2+ uptake. This study shows that the presence of L(+)-lactate does not inhibit excitation-contraction coupling in mechanically skinned fibres. PMID- 11417215 TI - A novel mechanism associated with idiopathic ventricular fibrillation (IVF) mutations R1232W and T1620M in human cardiac sodium channels. AB - Two mutations associated with idiopathic ventricular fibrillation (IVF) are localized within extracellular loops between segments DIIIS1-S2 (R1232W) and DIVS3-S4 (T1620M) of the human cardiac sodium channel (hNav1.5) alpha-subunit. We studied wild-type hNav1.5 channels and hNav1.5 channels with the R1232W/T1620M double mutation expressed in Xenopus oocytes using the cell-attached macropatch technique. We demonstrate that these mutations destabilize the fast-inactivated state (described with a two-state first-order reaction model) by decreasing reaction valence, accelerating recovery, and slowing the onset of fast inactivation, collectively resulting in delayed decay of macroscopic currents. R1232W/T1620M mutations in hNav1.5 channels also significantly increase steady state channel availability, indicating that mutated channels occupy the slow inactivated state less than hNav1.5 channels. Under the stress of repetitive depolarizing pulses, R1232W/T1620M channels demonstrate less use-dependent current reduction compared to wild-type channels. We propose that increased channel availability coupled with destabilized fast inactivation contributes to the pathological effect of R1232W/T1620M mutations, and leads to increased excitability of cardiac tissue in vivo. PMID- 11417216 TI - Different regulation of left ventricular ANP, BNP and adrenomedullin mRNA in the two-kidney, one-clip model of renovascular hypertension. AB - The aim of our study was to clarify whether atrial (ANP) and brain (BNP) natriuretic peptides and the hypotensive peptide adrenomedullin (ADM) are regulated differently in the rat heart in the two-kidney, one-clip model of renovascular hypertension. We assessed messenger ribonucleic acid (mRNA) abundance and distribution of ANP, BNP and ADM in the ventricles and atria of rats after unilateral renal artery stenosis (clipping). Rats were clipped for 6 h or 1, 2 or 4 days and mRNA levels were assessed semiquantitatively in left and right atria and ventricles by RNase protection assay. Left ventricular BNP mRNA up-regulation (4.3-fold after 6 hours) preceded ANP up-regulation (4.5-fold after 1 day) and seemed to be transient, whereas ANP mRNA levels were still elevated at day 4 (2.4-fold vs. sham). The right ventricle and the atria did not participate in these responses. Despite the massive changes of natriuretic peptide mRNAs, ADM mRNA did not change in either the ventricles or the atria. In contrast to ANP and BNP mRNA, which predominate in atrial tissue, mRNA for adrenomedullin is equally distributed in ventricles and atria. Plasma levels of immunoreactive (ir)-ANP and ir-BNP changed in parallel with left ventricular mRNA levels. Our findings suggest that renovascular hypertension induced by clipping the renal artery leads to immediate, but independent, up-regulation of ANP and BNP mRNA in the left ventricle whereas adrenomedullin mRNA is not changed. PMID- 11417217 TI - Influence of dehydration on glycerophosphorylcholine and choline distribution along the rat nephron. AB - Glycerophosphorylcholine is one of the four major organic osmolytes in renal medullary cells, changing their intracellular osmolyte concentration in parallel with extracellular tonicity during cellular osmoadaptation. In this study, the tubular content of glycerophosphorylcholine was quantified in untreated and 48-h dehydrated male rats. A chemiluminescence ultra-micromethod was developed to measure choline at the picomolar level in single tubules microdissected from collagenase-treated kidneys. The glycerophosphorylcholine level was calculated as the difference between total choline after acid hydrolysis and the free tubular choline content. In accordance with the glycerophosphorylcholine distribution pattern in different renal zones of untreated rats, low amounts of glycerophosphorylcholine were found in all cortical and outer medullary structures (< 35 pmol/mm), whereas increasing amounts were detected towards the papillary tip (163 pmol/mm). As a percentage of total choline, the level of free tubular choline varied from 4.2% in outer medullary proximal tubules to 30.3% in the inner medullary collecting ducts adjacent to the outer medulla (IMCD1). Antidiuresis led to a nearly twofold increase in glycerophosphorylcholine content in papillary collecting ducts. The osmolality-dependent regulation of organic osmolytes in single microdissected tubules has been demonstrated for the first time. Furthermore, the high tubular glycerophosphorylcholine concentration compared to sorbitol and myo-inositol emphasizes the predominance of glycerophosphorylcholine in the inner medulla and papilla of the rat kidney. PMID- 11417218 TI - Dose-dependent, prion protein (PrP)-mediated facilitation of excitatory synaptic transmission in the mouse hippocampus. AB - Disruption of both alleles of the prion protein gene, Prnp, has been shown repeatedly to abolish the susceptibility of mice to developing prion diseases. However, conflicting results have been obtained from phenotypic analyses of prion protein (PrP)-deficient (Prnp0/0) mice lines. To explore the possible neurophysiological properties associated with expression or absence of the normal isoform of the cellular prion protein (PrPC), we used conventional in vitro extracellular field potential recordings in the hippocampal CA1 area of mice from two independently-derived Prnp0/0 strains. Basal synaptic transmission and a short-term form of synaptic plasticity were analysed in this study. Results were compared with animals carrying a wild-type mouse PrP transgene to investigate whether PrP expression levels influence glutamatergic synaptic transmission in the hippocampus. There was a clear correlation between excitatory synaptic transmission and PrP expression; i.e. the range of synaptic responses increased with the level of PrPC expression. On the other hand, the probability of transmitter release, as assessed by paired-pulse facilitation, appeared unchanged. Interestingly, whereas the overall range for synaptic responses was still greater in older mice over-expressing PrPC, this effect in these animals appeared to be due to better recruitment of fibres rather than facilitation of synaptic transmission per se. Taken together, these data are strong evidence for a functional role for PrPC in modulating synaptic transmission. PMID- 11417219 TI - An apical membrane Na+/H+ exchanger isoform, NHE-3, is present in the rat epididymal epithelium. AB - An acidic milieu is required for sperm maturation and for keeping sperm quiescent during storage in the cauda epididymidis. Previous studies have implicated a Na+/H+ exchanger (NHE) in epididymal acidification together with carbonic anhydrase (CA) and vacuolar proton adenosine triphosphatase (H(+)-ATPase). The present studies were undertaken to discover whether the NHE isoform involved is NHE-3, which is known to mediate Na+ and HCO3- absorption in renal tubules. Using the reverse transcription polymerase chain reaction technique (RT-PCR), Northern blot analysis and in situ hybridization, NHE-3 mRNA was detected mainly in the cauda epididymis and to a lesser extent in other regions of the epididymis. Immunohistochemical studies showed that NHE-3 was present in the apical membranes of the epithelial principal cells and confirmed that its expression is strongest in the cauda region, decreasing towards the more proximal regions. Immunoblotting showed a similar expression pattern. These results demonstrate that NHE-3 is expressed in the rat epididymal duct with strongest expression in its cauda region. These findings are thus consistent with the possibility that NHE-3 in the epididymal duct is involved in luminal Na+ and/or HCO3- absorption, as in the renal proximal tubule, and thereby in the regulation of sperm motility and maturation. PMID- 11417220 TI - Modulation of the epithelial Ca2+ channel ECaC by extracellular pH. AB - We investigated the effect of extracellular pH on whole-cell currents through the epithelial Ca2+ channel, ECaC, expressed in HEK 293 cells. Both mono- and divalent current densities were significantly smaller at pH 6.0 than at pH 7.4. At pH 8.5 they were slightly larger. Lowering extracellular pH enhanced the slow component of monovalent current activation at negative potentials but had no significant effect on the kinetics of Ca2+ currents. The kinetics of block of monovalent cation current by extracellular Mg2+ was significantly changed at high and low pH. The time constant of the time- and voltage-dependent current component during a voltage step to -140 mV was significantly larger at pH 8.5 than at pH 7.4. At pH 6.0 it was almost absent. The [Mg2+] inhibiting 50% of monovalent current through ECaC at pH 6.0 (IC50) was 323 +/- 23 microM (n = 8), compared with 62 +/- 9 microM (n = 4) at pH 7.4 and 38 +/- 4 microM (n = 8) at pH 8.5. The affinity of ECaC for Ca2+ was also affected by extracellular pH, shifting from 4.8 +/- 0.7 microM (n = 6) at pH 6.0 to 161 +/- 30 nM (n = 5) at pH 7.4 and 425 +/- 117 nM (n = 8) at pH 8.5. PMID- 11417221 TI - Membrane cross-talk in the early distal tubule segment of frog kidney: role of calcium stores and chloride. AB - The activities of transport mechanisms in epithelial cells are generally coordinated in order to minimise disturbances in cellular ion content and volume. Furosemide, a potent inhibitor of transport in the renal diluting segment, up regulates apical K+ channel activity following the release of calcium from intracellular stores. The signal pathway between furosemide application and this calcium release is not known. Single early distal tubule segments from frog kidney were permeabilised with saponin in order to monitor calcium levels within cytoplasmic stores using the calcium-sensitive dye, mag-fura. The uptake (or release) of calcium to (or from) stores was initiated by adding agents to the bath solution, which is in direct contact with the intracellular organelles. ATP promoted calcium uptake into stores, whereas ATP removal led to a slower, spontaneous calcium release. Following loading, calcium stores could be rapidly depleted by inositol 1,4,5-trisphosphate (IP3), but not ryanodine. Calcium release was evident upon lowering the "intracellular" chloride concentration from 12 to 4 mM, equivalent to the fall in chloride induced by furosemide in intact cells. These results suggest that intracellular chloride may function as a second messenger, mediating cross-talk between the apical membrane and intracellular calcium stores. PMID- 11417222 TI - Excessive glucocorticoid exposure during late intrauterine development modulates the expression of cardiac uncoupling proteins in adult hypertensive male offspring. AB - We investigated the impact of intrauterine growth retardation and fetal programming of hypertension by maternal dexamethasone treatment on cardiac uncoupling protein (UCP) expression during development and in adulthood in the rat. Dexamethasone administered via an indwelling osmotic pump (100 micrograms/kg body mass per day from day 15 of gestation) decreased fetal body mass at day 21 of gestation (by 13%; P < 0.05), elicited significant (+24%, P < 0.01) systolic hypertension and elevated corticosterone levels (+15%; P < 0.05) in adult (24 week-old) male offspring. Cardiac UCP-2 and UCP-3 protein expression was significantly upregulated during early postnatal development, reaching 1.7-fold and 2.7-fold the respective fetal day-21 levels by postnatal day 7 and reaching plateaus at postnatal days 15-21 (2.5-fold and 3.5-fold of respective fetal levels). Cardiac UCP protein expression at fetal day 21 and the ontogeny of cardiac UCP expression during early postnatal life were unaffected by prenatal dexamethasone treatment. Prenatal dexamethasone treatment did not abrogate the postnatal surge in corticosterone levels or modify plasma non-esterified fatty acid (NEFA) levels over this period. However, UCP-2 and UCP-3 protein expression was significantly downregulated in the hearts of adult hypertensive male offspring of dexamethasone-treated mothers (to 27% and 65% of control values respectively). We propose that changes in cardiac UCP protein expression are linked with changes in cardiac metabolic fuel selection (from glucose-->fatty acids at birth and from fatty acids-->glucose during hypertension). PMID- 11417223 TI - HCO3- potentiates the cAMP-dependent secretory response of the human distal colon through a DIDS-sensitive pathway. AB - We used the Ussing short-circuit technique to investigate the role of HCO3- in the adenosine 3',5'-cyclic monophosphate (cAMP)-dependent secretory response of the human distal colon. In HCO3(-)-free 4-(2-hydroxyethyl)-1 piperazineethanesulphonic acid (HEPES)-Ringer's, forskolin (10 mumol l-1 mucosal and serosal) evoked a sustained increase in short-circuit current (Isc) (delta Isc = 24 +/- 3 microA cm-2, n = 57). However, this was only 25% of the forskolin stimulated Isc response in HCO3(-)-Ringer's (delta Isc = 84 +/- 8 microA cm-2, n = 57). The reduced response to forskolin in HCO3(-)-free HEPES-Ringer's was not due to inhibition of the secretory mechanism by HEPES, as replacing HCO3- with a different buffer, N-tris[hydroxymethyl)methyl-2-aminoethanesulphonic acid (TES), had a similar effect and inclusion of HEPES in the HCO3(-)-Ringer's did not reduce the secretory response. Similarly, it was not due to an indirect modulation of electrogenic Cl- secretion, as the forskolin-stimulated bumetanide sensitive Isc was comparable in the two Ringer's. Rather it was due to the activation of a HCO3(-)-dependent Isc which was inhibited by serosal 4,4' diisothiocyano-stilbene-2,2'-disulphonate (DIDS). This DIDS-sensitive Isc was not inhibited by acetazolamide, but it was inhibited by the replacement of bathing solution Cl- with gluconate, suggesting a role for a Na(+)-dependent Cl-/HCO3- exchanger in the cAMP-dependent secretory response of the human distal colon. PMID- 11417224 TI - Bursting activity in leech Retzius neurons induced by low external chloride. AB - We have investigated the bursting activity of Retzius neurons in the central nervous system of the leech Hirudo medicinalis as induced in Cl(-)-free saline by measuring membrane potential, membrane current and the intracellular calcium concentration ([Ca2+]i), using fura-2 or Oregon-Green488-Bapta-1. The Retzius neurons changed their low tonic firing to rhythmical bursting activity when the extracellular Cl- concentration ([Cl-]o) was lowered to 1 mM or less. In Cl(-) free saline (Cl- exchanged by gluconate), bursting was accompanied by a rise in intracellular Ca2+ in both cell body and axon, which oscillated in synchrony with the bursts. The Ca2+ transients depended on the amplitude and duration of the depolarization underlying the burst, and were presumably due to Ca2+ influx through voltage-dependent Ca2+ channels. In Ca(2+)-free, EGTA-buffered saline or in the presence of Ca2+ channel blockers verapamil (1 mM) or diltiazem (500 microM) the depolarizations underlying the bursts in Cl(-)-free saline were enhanced in amplitude and duration. Bursting was not affected by depleting the intracellular Ca2+ stores with cyclopiazonic acid. The depolarization in Cl(-)- and Ca(2+)-free saline did not evoke intracellular Ca2+ changes. The burst underlying membrane depolarization induced by Cl- removal was found to be due to a Na(+)-dependent persistent inward current and could be inhibited by saxitoxin (25-50 microM). The results suggest that a persistent Na+ current is generated in Cl(-)-free saline and induces the depolarization underlying rhythmic activity, and that presumably Ca(2+)-induced K+ currents modulate the bursting behaviour. PMID- 11417225 TI - Ca(2+)-activated Cl- channels in Ehrlich ascites tumor cells are distinct from mCLCA1, 2 and 3. AB - Using the whole-cell patch-clamp technique, we have studied the electrophysiological and pharmacological properties of the Ca(2+)-activated Cl- current present in Ehrlich cells. The currents activated slowly upon depolarization, deactivated upon hyperpolarization, and showed strong outward rectification. An increase in [Ca2+]i activated the current with an EC50 of 165.2 nM. Extracellular application of niflumic acid (100 microM) rapidly blocked the current in a voltage-dependent manner whereas sulfhydryl-modifying agents such as dithiothreitol (DTT, 1-2 mM) and N-ethylmaleimide (NEM, 100 microM) had no effect on Ca(2+)-activated currents in Ehrlich cells. Members of the recently discovered CLCA gene family are the only molecular candidates for Ca(2+)-activated Cl- channels cloned so far. Using RT-PCR we demonstrated that the appearance of a Ca(2+)-activated Cl- current in Ehrlich cells is not associated with the expression of the murine members of the CLCA family (mCLCA1-mCLCA3). Correspondingly, the kinetic and pharmacological properties of the Ca(2+) activated current in Ehrlich cells differ from those of CLCA-associated currents, which are time independent and DTT sensitive. Thus, phenotypic differences in combination with RT-PCR data point to the existence of different molecular species for Ca(2+)-activated Cl- channels. PMID- 11417226 TI - The C-terminal part of the R-domain, but not the PDZ binding motif, of CFTR is involved in interaction with Ca(2+)-activated Cl- channels. AB - Expression of the cystic fibrosis transmembrane conductance regulator (CFTR) inhibits Ca(2+)-activated Cl- channels (CaCC) by an unknown mechanism. This inhibition does not require CFTR activation (activity-independent inhibition), but is potentiated when CFTR is activated (activity-dependent inhibition). In this study, we evaluated, in endothelial cells, possible structural determinants for this interaction. Bovine pulmonary artery endothelium (CPAE) cells, which do not express CFTR, were transfected transiently with three hybrid CFTR constructs. The functional interaction between CaCC and CFTR was assessed using the patch clamp technique in the whole-cell configuration. CaCC was stimulated by application of adenosine 5'-triphosphate (ATP) to the bath solution. CFTR currents were evoked by application of a forskolin/3-isobutyl-l-methylxanthine (IBMX) cocktail. The inhibitory effect of CFTR was conserved when the PDZ (PSD 95/Discs large/ZO-1) binding motif was deleted (CFTR-delta PDZ). In contrast, both the CFTR activity-independent and -dependent inhibition of CaCC were abolished when the C-terminal part of the regulatory (R)-domain of CFTR was deleted (CFTR-delta R780-830). The activity-dependent inhibition of CaCC, but not the activity-independent inhibition, could be rescued by introducing the multiple drug resistance (MDR)-1 mini-linker in place of the deletion (CFTR-delta R linker). It is concluded that the C-terminal part of the R-domain is an important determinant for CFTR-CaCC interaction. PMID- 11417227 TI - Identification and characterisation of human xCT that co-expresses, with 4F2 heavy chain, the amino acid transport activity system xc-. AB - We have identified a new human complementary deoxyribonucleic acid (cDNA), for the xc- amino acid transporter (HGMW-approved name SLC7A11; also known as human xCT), that, when co-expressed with the heavy chain of surface antigen 4F2 (4F2hc, also termed CD98), but not with rBAT, (related to the bo,+ amino acid transporter), induces system xc- transport activity in Xenopus oocytes. Human xCT is the seventh human member of the family of amino acid transporters that are subunits of 4F2hc or rBAT and, inview of its amino acid sequence identity (89%) with mouse xCT, is most probably the human orthologue thereof. The amino acid transport activity induced by the co-expression of human 4F2hc and xCT in Xenopus oocytes was sodium independent and specific for L-cystine, L-glutamate and L aspartate. This activity also functioned in an exchange mode (e.g. cystine/glutamate) with a substrate stoichiometry of 1:1. Expression of human xCT alone in oocytes did not induce amino acid transport activity and the expressed xCT protein was localised intracellularly. When human xCT was co-expressed with 4F2hc, the former localised to the oocyte plasma membrane. Tissue-expression studies showed that human SLC7A11 mRNA is expressed mainly in the brain, but also in pancreas and in cultured cell lines. The transport characteristics of human xCT and the distribution of its tissue expression strongly suggest that it corresponds to the human amino acid transporter system xc-. PMID- 11417228 TI - Changes in element composition of A6 cells following hypotonic stress. AB - Cellular element concentrations and dry weight contents were determined in A6 epithelia using electron microprobe analysis. This was done to assess the quantitative contributions of Na, K and Cl to the regulatory volume decrease (RVD) and isovolumetric regulation (IVR) after decreasing the basolateral osmolality from 260 to 140 mosmol/kg in a stepwise or gradual way. Two minutes after inducing acute hypotonic stress the cells behaved almost like ideal osmometers, as indicated by a pronounced increase in cell height and decreases in the cellular dry weight and concentrations of all measured elements by about the same degree. Sixty minutes after inducing acute hypotonic stress the dry weight and concentrations of the impermeant elements P, Mg and Ca had returned approximately to control values, indicating normalized cell volume. Na, K and Cl concentrations, however, remained greatly reduced. The cellular amounts of Na, K and Cl diminished during RVD by approximately 31%, 24% and 46%, respectively. The dry weights and element concentrations measured 60 min after inducing acute hypotonic stress were similar to those obtained after a continuous reduction of basolateral osmolality. The cellular loss of Na and K following hypotonic stress exceeded that of Cl by about 40 mmol/kg wet wt., suggesting the exit of an other anion and/or the titration of fixed negative charges. The contribution of Na, K and Cl to total cellular osmolality increased from about 75% under control conditions to about 85% during RVD and IVR. Since only approximately 70% of the loss of cellular osmolytes necessary for the observed RVD and IVR is accounted for by the cellular exit of Na, K and Cl, other osmolytes, possibly amino acids, must leave the cells following hypotonic stress. PMID- 11417229 TI - In situ assessment of shortening and lengthening contractile properties of hind limb ankle flexors in intact mice. AB - The availability of animal models with disrupted genes has increased the need for small-scale measurement devices. Recently, we developed an experimental device to assess in situ mechanical properties of isometric contractions of intact muscle complexes of the mouse. Although this apparatus provides valuable information on muscle mechanical performance, it is not appropriate for determining contractile properties during shortening and lengthening contractions. In the present study we therefore developed and evaluated an experimental apparatus for assessment of shortening and lengthening contractile properties of intact plantar and dorsal flexors of the mouse. The current through a custom-built, low-inertia servomotor was measured to assess contractile muscular torque ranging from -50 to mN.m. Evaluation of the fixation procedure of the animal to the apparatus via 3-D monitoring of the muscle-tendon complex length showed that the additional shortening in length due to a contraction with maximal torque output has only minor effects on the measured torque. Furthermore, misalignment of the axis of rotation of the apparatus relative to the axis of rotation in the ankle joint, i.e. eccentricity, during a routine experiment was estimated to be less than 1.0 mm and hence did not influence the measured torque output under our experimental conditions. Peak power per unit muscle mass (mean +/- SD) of intact dorsal and plantar flexors was 0.27 +/- 0.02 and 0.19 +/- 0.03 W.g-1, respectively. The angular velocity at maximal peak power generated by the dorsal flexor complex and the plantar flexor complex was 1100 +/- 190 and 700 +/- 90 degrees.s-1, respectively. PMID- 11417230 TI - Reconstitution of neurotransmission by determining communication between differentiated PC12 pheochromocytoma and HEL 92.1.7 erythroleukemia cells. AB - Neurotransmitter release was monitored using fura-2-loaded HEL 92.1.7 cells dispersed among differentiated PC12 cells (loaded with another Ca2+ indicator fluo-3) and immobilised using transparent polycarbonate membrane filters with uniform pore size. Depolarisation with K+ caused a rapid rise in Ca2+ concentration in the PC12 cells, followed by a delayed secondary Ca2+ response in simultaneously monitored nearby HEL cells. There was a lag period of about 20 s between the responses of the two cell types. Voltage-gated Ca2+ channels in PC12 cells were inhibited by the P/Q-type (omega-conotoxin MVIIC, omega-agatoxin IVA), N-type (omega-conotoxin GVIA) and L-type channel blockers (nifedipine) as determined using fura-2 or whole-cell patch-clamp recordings. The communication between the cell types on the other hand was sensitive to P/Q- and N-type but not to L-type channel blockers. This suggests that, as in neurons, P/Q- and N-type Ca2+ channels mediate the release of neurotransmitters acting on HEL cells. Theoretically, the procedure employed should be sensitive enough to detect single exocytotic events. Our results demonstrate that a random distribution between effector and target cells is sufficient to allow communication between cells in a manner similar to extrasynaptic transmission. PMID- 11417231 TI - [Hip prosthesis implantation--only good long-term results count!]. PMID- 11417232 TI - [Particle disease. Is tribology a topic in revision surgery?]. AB - To improve the longevity of endoprostheses, the main goal is to reduce wear. Polyethylene together with metal or ceramic is currently the most frequently used combination. Their clinical success is well documented in the literature. Many attempts to improve polyethylene in the past have failed. Materials successful in the laboratory have failed in clinical use. The most recent competitors of ultra high molecular weight polyethylene (UHMWPE) are the highly cross-linked polyethylenes (HCLPE) and the hard-on-hard couplings such as metal-on-metal or ceramic-on-ceramic. Advantages and downsides regarding particle generation and higher standards of precision in positioning the components are discussed. PMID- 11417233 TI - [Acetabulum revision. Classifications and treatment possibilities]. AB - Several different defect classifications have been published. The most commonly used are described. For clinical practice, differentiation between "contained" and "noncontained" or segmental defects has proven useful. If the acetabular rim provides support, press-fit cups can be used without screws. With an appropriate reaming technique, it is possible to create a sufficiently supportive bed for the implant in many cases. This allows the use of a press-fit acetabular component over a wide range of defects as long as a tilting of the cup can be avoided. Between 1988 and 1995, 439 acetabular cups were revised at the Orthopedic University Clinic in Basel, 171 of them using the Morscher press-fit cup. After a mean follow-up of 7.1 years, not a single cup had to be revised for aseptic loosening. The 9.5% dislocations were due to abductor insufficiencies because of trochanteric pathologies or muscular alterations from former approaches. PMID- 11417234 TI - [Hooked roof cup in revision of difficult loose hip prosthesis cups. Results after a minimum of 10 years]. AB - The long-term results of acetabular revision after total hip arthroplasty (THA) with the use of a reinforcement ring with hook were evaluated. The study included 57 cases of surgery dating back 10 years or more. Of a total of 54 patients (57 operated hips), 18 patients (19 hips) died during the 10-year period and 2 patients (2 hips) were completely lost to follow-up. At the time of the revision surgery, the mean age of the remaining 34 patients (36 hips) was 62.5 years (range: 47-80). A THA revision was done in 25 cases and an acetabular revision only in 11 cases. The most common acetabular defect was a combined segmental and cavitary defect (n = 19), and in three cases there was pelvic discontinuity. Autologous or homologous cancellous bone grafts were used to fill acetabular cavities in 17 hip joints. Structured bone grafts, predominantly homologous bone, were used in ten cases for acetabular reconstruction. At a mean follow-up of 11.4 years (range: 10-14.5) three hip joints (8%) had undergone further revision. The revisions were done for aseptic loosening of the acetabular component in two cases and a septic loosening of both components in one case. Three further cases (8%) revealed signs of acetabular loosening. Two of these three patients were symptomatic but refused further revision surgery. In the 33 unrevised hip joints, a good or excellent clinical result with a d'Aubinge score of more than 14 points was found in 30 cases (92%). Osseous acetabular reconstruction with the use of a reinforcement ring leads to favorable results compared to other techniques. In the authors' opinion, this technique is preferable to those using oversized cups without osseous reconstruction of the acetabulum. PMID- 11417235 TI - [Shaft revision. Classification and treatment]. AB - The goal of every hip revision is to restore as much function as possible. Frequent problems are defects of the load-carrying bony substance and functional deficits due to trochanteric avulsions, abductor insufficiencies, and lesions of the superior gluteal nerve. Disturbed functions of these important structures may lead to unsatisfactory results with high dislocation rates and limping. Various femoral defect classifications differ in their description of the lesion as well as in their proposed therapeutic approach. Significant differences exist in the field of allograft use. Among others, the AAOS, SO.F.C.O.T, and Paprosky's classifications are described. Our own easy to use alternative classification is introduced which takes into consideration the fixation principle of the new stem. It thus helps to select the type of implant and offers a guideline for technical modalities. A Type 1 defect allows a revision with a component suitable for a primary case. Type 2 defects can be treated with a short revision stem exceeding the length of the revised stem. Type 3 defects are treated with a "load-sharing" concept where an optimized metaphyseal fill and a cylindrical shape distally share the axial and torsion loads. Type 4 defects are reconstructed with a combination of a revision stem and an allograft. PMID- 11417236 TI - [Modular shafts in hip prosthesis revision surgery]. AB - The introduction of a proximally modular hip stem has greatly simplified difficult hip surgery. The long-term results of revision surgery indicate that the majority of revision cases can be handled with a proximal ingrowth implant, which helps speed bone recovery and simplifies re-revision if it is required. To date no particular downside has been noted as a result of the use of a modular stem. PMID- 11417237 TI - [The value of the Wagner SL revision prosthesis for bridging large femoral defects]. AB - Between 1988 and 1999 we used the Wagner SL revision prosthesis in 99 cases. The indications were aseptic and septic loosening, periprosthetic fractures, and Girdlestone situations. These were mostly Paprosky types 2 and 3 meta- and metadiaphyseal femoral bone defects. The intervention was the first revision in 49 cases and the second or up to the sixth revision in 50 cases. A transgluteal or transfemoral approach was usually chosen. We gradually reduced the anchorage area to a length of 8-12 cm. If after the first few postoperative months an osseous integration without radiolucency is achieved, a long-lasting integration can be expected. Due to considerable subsidence during the 1st year, six stems had to be replaced. Seven additional revisions were done because of hematoma and three because of seroma. According to the survival analysis, 92% of the stems remained in place after 10 years. No revision was executed between the 5th and 10th year. At the 1-year control, 96% of the patients were pain free and 90% were satisfied. Nevertheless, 80% limped and 33% used at least one crutch. Mainly because of deterioration of general health, the walking distance was considerably reduced between the 1st and the 5th postoperative year. There is little difference in the results after revision between a relatively small and an extended defect. At present, we limit the use of the SL revision stem mainly to extended defects. Careful planning of the operation is mandatory. PMID- 11417238 TI - [Intermediate term outcome of of a hip prosthesis revision system]. AB - The Zweymuller shaft for uncemented total hip arthroplasty was developed in the early 1970s. Encouraged by the clinical results with this stem, which was mainly used in primary arthroplasty, longer fitting stems were added to the line to accommodate bony defects and to allow for an optimal load transfer from proximal to distal. The principal of the design is to allow an optimal distal fixation while allowing the bone to remodel in the proximal part. This study reports on 89 patients who underwent revision surgery of the hip for mostly aseptic loosening. The results after a median follow-up of 36 months show an increase of the modified Harris hip score from 52 points pre- to 82 points postoperatively. Radiographic subsidence was found in nine cases, with eight having progressed for more then 3 mm. Postoperative complications occurred in 11.2%, with seven dislocations. Open revision became necessary in two cases. The stem reviewed here seems to achieve predictable results in cases where a proximal cone is still present to facilitate load transfer, while at the same time the quadrangular stem provides solid distal fixation and ensures rotary stability. PMID- 11417239 TI - [Cement removal with ultrasound in revision or total hip prosthesis]. AB - Bone cement (polymethylmethacrylate, PMMA) removal during revision hip arthroplasty can be a tedious, time-consuming process. The usual methods of removing cement include high-speed drills, chisels, saws, and reamers, which are often associated with fracture or perforation of the femoral shaft. An alternative very promising method is cement removal with ultrasound. We present an ultrasonic device for rapid cement removal with minimum risk of trauma to the fragile femoral bone. The technique of cement removal with ultrasound is described and problems and risks are addressed. PMID- 11417240 TI - [Long-term results with the cement-free Spotorno CLS shaft]. AB - This retrospective study reviews 298 sequential cementless CLS Spotorno stems, which achieved their primary fixation and bony in- or ongrowth predominantly in the metaphyseal region. The minimum follow-up was 10 years and the average follow up 11.7 years (range 10-14). The mean age at surgery was 55.2 years (range 32.2 68.9). Of the patients, 76.2% (227 stems) were followed up: 172 patients (57.7%) underwent clinical and radiological examination and 52 patients (55 stems, 18.5%) were interviewed by telephone. Of 298 stems, 13 (4.4%) had been replaced in the mean time, only 6 stems (2.0%) because of an aseptic loosening. The evaluation included both clinical (Merle d'Aubigne and Harris hip score) and radiographic parameters (plain X-rays). The mean Merle d'Aubigne score was 16.7 points, and the mean Harris hip score was 94.3 points. Radiolucent lines were detected in 33.7%; 5.8% showed enlargement within the last 3 years. Focal osteolyses were present in 30.8% of patients; 2.9% had progressed within the last 3 years. At follow-up, one patient presented with new clinical and radiological evidence of aseptic stem loosening. The current study showed an aseptic loosening of the CLS stem in 2.3%, a good osseointegration and good functional score results in more than 90%. The primary fixation and bony in- or ongrowth predominantly in the metaphyseal region seems to achieve good clinical results. Wear is the main reason for the osteolytic changes. Frequent clinical and radiological follow-up is important to detect bone reactions and loosening of the stem. PMID- 11417241 TI - [3-dimensional biomechanical study of a new flexible lumbar intervertebral disk implant]. AB - The aim of this study was to determine the significance of a new artificial nucleus implant for lumbar discs (polymethyl siloxane polymer, DR-PMSO) with special regard to the biomechanical properties of the spinal motion segment. The lumbar segments L1/L2 and L4/L5 were harvested from six calf cadavers. The segments were fixed on a special device mounted on a three-dimensional testing machine. Physiological load with a maximum of 7 Nm was applied in flexion/extension, right and left lateral bending, and right and left torsion. The movement of the measuring point (center of L1 and L4) was registered along three axes (sagittal, frontal, and vertical). Three cycles of measurements were performed on all specimens: (1) intact segment, (2) segment after nucleotomy, and (3) segment with PMSO. There was a statistically significant (p = 0.0313) increase in segmental mobility in all directions after nucleotomy with an increased mobility of the segment up to 30% for rotation and 50% for translation. After introduction of the PMSO implant, segmental mobility for all movement directions was restored with no statistically significant difference from the intact situation before nucleotomy. The disc material was found to be noncytotoxic according to current ISO 10993 standards. PMID- 11417242 TI - [North German Orthopedics Meeting. Berlin, 22-24 June 2000]. PMID- 11417243 TI - [Quality management in hip and knee prosthesis implantation]. PMID- 11417244 TI - [Detection of humoral immune response to inner ear proteins in patients with sensorineural hearing loss]. AB - BACKGROUND: The precise mechanism of inner ear disease is still unknown. An autoimmune reaction could be one of several possible pathogenic factors involved in progressive sensorineural hearing loss. Heat shock protein 70 is suggested to play an important role in the development of autoimmune diseases. The aim of this study is the investigation of humoral immune reactivity to inner ear components in patients with sensorineural hearing loss. METHODS: The presence of antibodies to inner ear components was determined by immuno-blotting extracted bovine or human inner ear proteins. Study groups consisted of patients with idiopathic progressive sensorineural hearing loss (group A), patients with Meniere's disease (group B), patients with sudden hearing loss (group C), patients with otosclerosis (group D), patients with Cogan's disease (group E), and individuals without hearing problems (group F). RESULTS: 40% of the patients with progressive sensorineural hearing loss showed reactivity against a 68-kDa protein extracted from bovine inner ear. In contrast to this, only 5% of healthy individuals and 10% with Meniere's disease showed reactivity against the 68-kDa protein from bovine inner ear or against bovine heat shock protein 70. Some of the patients who showed reactivity against bovine inner ear proteins were tested with human inner ear and human heat shock protein 70; all of these showed reactivity. Approximately 6% of the patients with sudden hearing loss (group C), otosclerosis (group D), and Cogan's disease (group E) showed reactivity to inner ear proteins. A non-specific humoral immune reaction against inner ear proteins with molecular weights of 30, 40, 50, 60, and 220 kDa was observed in all patients. DISCUSSION: These results indicate a humoral immune reactivity against heat shock protein 70, which might be responsible for the pathogenesis of progressive sensorineural hearing loss. PMID- 11417245 TI - [Management of a deaf-blind patient with a 12-channel cochlear implant]. AB - A case study of a deaf-blind patient implanted with the COMBI40+ cochlear implant is presented. The patient is a 70-year-old woman who has been blind since the age of 40. Since 1977 the hearing loss continuously increased. Before surgery she had a very small residual hearing (with hearing aid: 40% speech intelligibility by four syllable numbers) and she used Braille and finger-spelling on her hand to communicate with other people. The patient has a high intelligence and so she did not have any problems when the speechprocessor was fitted and during the hearing therapy. After one month she had an open-set-speech recognition. The quality of her life is much better than before surgery. The communication by finger-spelling on her hand is not necessary any longer. PMID- 11417246 TI - [Cardiotoxicity of 5-fluorouracil. Clinical relevance of palliative radiochemotherapy of malignant head-neck tumors]. AB - BACKGROUND: In the past cardiotoxicity has been increasingly reported as 5 fluorouracil is widely used as well in curative adjuvant as in palliative chemotherapy of malignant neoplasms. METHODS: We report on two cases of 5 fluorouracil-associated cardiotoxicity on palliative treatment of head and neck cancer and evaluate the importance of intensive drug monitoring. RESULTS: 5 fluorouracil-associated cardiotoxicity occurs with an incidence of 1.1-4.5%. Ischaemic cardiopathy due to vasospasms is believed to be the most common manifestation. CONCLUSIONS: Since most patients with head and neck cancer have risk factors for cardiovascular disease, 5-fluorouracil-associated cardiotoxicity should not be underestimated and patients should be monitored carefully. PMID- 11417247 TI - [Mucoepidermoid carcinoma of the salivary glands after high dosage radiotherapy]. AB - BACKGROUND: High-dose radioiodine therapy following total thyroidectomy is standard for patients suffering from differentiated thyroid carcinoma and contributes significantly to their favourable prognosis. Due to active iodine accumulation, high focal radiation doses are received by the salivary glands. PATIENTS/RESULT: Report on two patients, who received multiple high-dose radioiodine treatments because of a differentiated metastatic thyroid carcinoma. A few years later, they developed a mucoepidermoid carcinoma of the salivary glands. Due to the high cumulative radiation dose, radiation-induced secondary malignancies following radiation-induced sialadenitis appears likely, although no causal connection could be proven. CONCLUSION: Consistent protection of the salivary glands during radioiodine therapy as well as the follow-up of the many long-term survivors of differentiated thyroid carcinomas is desirable to further lower the salivary gland-related side effects and to detect secondary malignancies as early as possible. PMID- 11417248 TI - [Immunohistochemical detection of hormones in hypophyseal adenomas of the rat after fractionated irradiation]. AB - BACKGROUND: The aim of this study was to determine the hormones produced in pituitary gland tumours (TM) following fractionated external irradiation in rats. The TM arose in the course of studies on other questions of radiation effects. METHODS: The left neck and skull base of sixty female Wistar rats, 3 to 4 months of age at the beginning of the external irradiation, were subjected to roentgen rays exposure, fractionated to 2 Grays daily, either up to 20, 40 or 60 Gy. The midline of the neck and skull was always inside the radiation field. The animals were sacrificed either 6 months or 12 months after completion of the randomly assigned irradiation protocol. Five non-irradiated rats served as controls. RESULTS: No TM developed in the non-irradiated animals. Out of 60 irradiated rats 9 developed a pituitary TM (15%). In remnants of the normal adenohypophysis we revealed immunohistochemical reactivity for growth hormone (GH), adrenocorticotropic hormone (ACTH), prolactin (PRL), thyroid stimulating hormone (TSH), and follicle stimulating hormone (FSH). In the radiation-induced adenomas, 4 TM were immuno-reactive for TSH, 4 for GH, 2 for PRL, and 2 for FSH. The TM did not react with anti-ACTH and anti-LH (luteinizing hormone) anti-sera. CONCLUSIONS: Obviously, irradiation accelerates the development of pituitary gland adenomas. In this series the TM were predominantly incidental findings at necropsies after pretermed follow-up intervals with a variety of hormone producing cells. On the other hand, four of nine TM were null adenomas. Necropsies of the brain should be carried out routinely in irradiation studies of the skull. PMID- 11417249 TI - [Rational diagnosis of pediatric pharyngeal abscess]. AB - BACKGROUND: Diagnostic procedures intend to differentiate superficial cervical abscesses from deep abscesses and uncomplicated lymphadenitis. They should provide identification of the causing agent, of the route of infection and of possible underlying diseases. Present techniques for examining pediatric cervical abscesses should be assessed. METHODS: Currently published data on diagnostic procedures in pediatric neck abscess from National Library of Medicine data files were evaluated. Moreover, clinical findings, diagnostic procedures, therapy and outcome of 47 children with cervical abscess treated from 1992-1996 were retrospectively evaluated. RESULTS: Studies providing appropriate evidence on the value of various diagnostic procedures in pediatric cervical abscesses were not found. In 47 patients with confirmed cervical abscess treated at our department, clinical examination, erythrocyte sedimentation rate and ultrasound examination of the neck were sufficient to establish the correct diagnosis. Additional investigations such as MRI or CT-scans, chest X-rays, Mendel-Mantoux test, various virus titres and other laboratory examinations were performed only in selected cases. The route of invasion (tonsil, dental or otogeneous) could be identified in 13 children. In 4 children an infected cervical cyst was found. CONCLUSIONS: Clinical examination, erythrocyte sedimentation rate and sonography are appropriate to establish the diagnosis of pediatric cervical abscess and in accordance with the principles of Managed Care. PMID- 11417250 TI - [Early surgical pharyngostoma in therapy of postoperative pharyngeal fistulas]. AB - BACKGROUND: Postoperative salivary fistulas after laryngectomy occur in 2-65% of cases according to the literature. They are not only inconvenient but also dangerous, as the permanent leakage of saliva prevents wound healing and may cause a life threatening arrosion hemorrhage from the jugular vein or carotid artery. Spontaneous closure of such fistulas is unlikely unless the leakage of saliva can be reduced sufficiently by suction and wound compression. However, about 50% of these patients develop a mucocutaneous pharyngostome after several weeks or even months, then requiring extensive flap reconstruction. In order to shorten this long rehabilitation process and to protect the patient from the danger of hemorrhage the commonly recommended therapeutic concept was modified in that the saliva was completely drained from the wound by creation of a submental epithelialized pharyngostome. In this paper our 10 year experience with this concept is reported. PATIENTS AND METHOD: During the period from 1991 to 1999 postoperative salivary fistulas were seen in 9 of 94 consecutive patients after total laryngectomy. Right after diagnosis a meticulous surgical wound debridement was performed and a submental pharyngostome was created in each case. After healing of the mucocutaneous lining the pharyngostome itself was closed with regional skin flaps. RESULTS: In 7 of the 9 patients a quick and permanent closure of the fistula was achieved by the treatment described. The sooner after diagnosis of the fistula the pharyngostome was created, the earlier the closure of the fistula was obtained. In 1 patient the pharyngeal reconstruction was complicated by nosocomial infections, 1 patient died in the course of treatment due to non fistula related concomitant disease. CONCLUSIONS: In case of postoperative salivary fistulas following laryngectomy the immediate creation of a mucocutaneous pharyngostome can reduce the duration of treatment and at the same time protect the patient from the danger of vascular arrosion. PMID- 11417251 TI - [Osteoma of the maxillary sinus]. AB - BACKGROUND: Osteoma of the paranasal sinuses is a rare and benign entity that develops slowly. If they occur, locations within the frontal sinus and ethmoid cells are more frequent, whereas osteomas in the sphenoid or maxillary sinus are very rare. CASE: We report on a 25-year old female patient presenting with a bony mass in the right maxillary sinus. A standard external approach using a modified Caldwell-Luc procedure was successfully employed as endoscopic endonasal surgery was limited by the size of the bony lesion. CONCLUSIONS: Trauma or infection are often suggested as an etiologic factor in the pathogenesis of osteoma. In our case the development of maxillary osteoma was related to dental extraction and postoperative fistula. External standard procedures remain mandatory if endonasal endoscopic surgery fails in the removal of osteoma of the paranasal sinuses. PMID- 11417252 TI - [Combined surgical and prosthetic therapy of severe obstructive sleep apnea--a case report]. AB - We report on a 41-year old patient who complained of loud snoring, excessive daytime sleepiness and chronic nasal obstruction. Clinical findings were septal deviation and enlarged turbinates, tonsillar hypertrophy with velar webbing and pharyngeal narrowing. Polysomnography revealed severe obstructive sleep apnoea syndrome with an apnoea-hypopnoea index (AHI) of 51.7/h. As the patient refused nCPAP therapy, we performed septoplasty with conchotomy and an uvulopalatopharyngoplasty with tonsillectomy. Snoring and excessive daytime sleepiness disappeared completely and the AHI decreased to 31.1/h. The mandibular advancement device Snorban was subsequently fitted. We found a complete resolution of OSAS. The AHI was 4.4/h. The postsurgical polysomnographic results were stable two years after surgery. However, the patient discontinued using the oral device as he did not feel any additional benefit when using it. The combination of UPPP and mandibular advancement device can resolve a severe OSAS. PMID- 11417253 TI - [Endoscopically controlled hemostasis in tonsillectomy]. PMID- 11417254 TI - [Diagnosis and therapy of laryngeal diseases in the history of medicine. I: The pre-laryngoscopic era]. AB - BACKGROUND: ANATOMY, PHYSIOLOGY AND PATHOLOGY: Hippocrates and Aristotle did not yet have a clear idea of the anatomy and physiology of the larynx. 500 years later Galenos carried out subtle studies on animals and elaborated a fairly precise description of the different cartilages, muscles and nerves of the larynx; he was the first to demonstrate the superior and the recurrent nerves of the larynx. He performed numerous experiments on animals with temporary or permanent interruption of the nervous function and thus developed a correct opinion of the vocal physiology. However, he did not draw clinical conclusions from his knowledge. In the 16th century the macroscopic anatomy of the larynx was elaborated further bei Andreas Vesalius, Fabricius ab Aquapendente et al.; Jacob Henle in 1838 described the different epithelia in the larynx, based on microscopic studies. The pathology was inaugurated by Giovanni-Battista Morgagni in 1761, including the pathology of the larynx. Francis Home in Edinburgh (1765) and Pierre-Fidele Bretonneau in Tours (1826) made the first studies on croup and diphtheria and coined these names. Fr. Hermann Albers in Bonn in 1829 published the first monography on diseases of the larynx, incorporating more than 80 relevant case reports from the literature and numerous observations of his own. The first system of the pathology of the larynx was put up by von Rokitansky in Vienna. DIAGNOSIS AND THERAPY OF DISEASES OF THE LARYNX: Before the invention of laryngoscopy the physicians could deduce the underlying disease only from the clinical symptoms like hoarseness, dyspnoea, dysphagia. The conservative therapy followed the theory of humours and included bloodletting, leeches, purgatives and emetics; the surgical procedures were blind caustery with silver nitrate and ablation of polyps guided by palpation and scarification with a knife in cases of oedema; in desperate dyspnoea endonasal intubation was attempted (Desault 1789 1803) or a tracheotomy performed. Concluding the chapters a typical patient history of 1813 with a case of purulent laryngitis where most of these measures were applied but could not avert but rather provoked the death of the patient. This historical development is described with numerous anecdotic details. PMID- 11417255 TI - [Interesting case no. 45. Pharyngeal manifestation of chondroid chordoma]. AB - Chordomas usually occur in the axial skeleton and they arise from the remnants of notochord. Their growth is slow and they often metastase. Most frequently chordomas are situated in sacral bone (50%), in spheno-occipital region of the skull base (35%) and in the cervical and lumbar spine (15%). We reported an unusual case of chordoma of the cervical spine with participation of the parapharyngeal space. The cervical chordoma was excised by total resection of the tumor mass with partial vertebrectomy C2/C3. PMID- 11417256 TI - [Molecular biology studies of the effect of genetic factors in disease susceptibility on course of illness in patients with squamous epithelial carcinomas of the head-neck area]. PMID- 11417257 TI - [Traumatology of the soft tissues of the neck. I]. PMID- 11417258 TI - [Mental health care at the turn of the millennium--on the way toward two-class psychiatry]. AB - Up to now the special position of psychiatry is determined by prejudices in connection with violence and dangerousness of the mentally ill. The reforms of mental health care since the fifties were directed not only at the improvement of the treatment and the situation of psychiatric patients, but also at the reduction of prejudices and stigmata--i.e. a normalisation of the aforementioned special position. However, in connection with the stepwise progress of the reforms one could notice an increasing neglect of a subgroup of severely mentally ill who are not able to accept the offers of modern mental health care, or whose care-requirements are not met by the current developments. This together with the withdrawal of psychiatry from all those aspects of treatment, dealing with aggressive behaviour of or coercion against mentally ill led to a stepwise removal of certain patients--recruited mainly from the aforementioned subgroup- from the general mental health care system. Initially this development could be observed only in the USA, but in the meantime several European countries are appearing to catch up. This reinforces the prejudices against mental patients in general which are present up to now, is counterproductive with regard to the intended aims of mental health reforms and increases the danger of undifferentiated and restrictive political reactions. PMID- 11417259 TI - [Imperative auditory hallucinations in schizophrenia]. AB - Command hallucinations are a subtype of auditory verbal hallucinations in schizophrenia. More likely than any other schizophrenic symptom, they may have an impact on the individual's behavior. In the forensic literature, aspects of dangerous actions induced by psychotic motivation have been discussed. From the psychiatric point of view, the main problem is the unpredictability of behavioral consequences. The present study examines psychopathological details of command hallucinations in 31 hospitalized schizophrenic patients, 10 females and 21 males, using a questionnaire including 24 items. The predictive accuracy of a distinct set of signs was rated concerning compliance vs. resistance of hallucinated commands. The predictors are: identifying the hallucinated voice, being affected by emotions after hallucinations, and misregarding the voice as being real. In the presence of these conditions, behavioral consequences are predicted by a value which was 6.7 times higher than that without any information about predictors. In the absence of these conditions, resistance of command voices is predicted by a value of 7. PMID- 11417260 TI - [Recurrence of paranoid hallucinatory psychoses after beginning a fasting period in a patient with anorexia nervosa]. AB - We report the case of a 37-year old female patient with anorexia nervosa. For many years, this patient fasted for various periods, from days to weeks, in order to reduce her body weight and to achieve an euphoric effect. In the course of the past four years she developed acute paranoid-hallucinatory psychosis immediately following one week episodes of fasting, requiring psychiatric in-patient treatment three times. Psychotic prodromal symptoms could not be elicited before nor at the beginning of fasting periods, pointing towards a caustic relationship between the fasting and the psychotic decompensation. Based on this case, we discuss the hypothesis that the fasting in anorexia nervosa leads to acute hyperactivity of the dopaminergic system which could explain the development of paranoid-hallucinatory psychosis. This case exemplifies that vulnerable patients with a similar history should strongly be advised to avoid fasting and other dietetic methods. Explanation of the enhanced risk to develop psychosis could be of therapeutic use for treating the anorexia nervosa in such patients. PMID- 11417261 TI - [Lycanthropy-zooanthropism--discussion of a psychopathological phenomenon]. AB - The belief to be transformed into an animal is named Lycanthropy. "Zooanthropismus" is the German equivalent. Three case reports raised the question how this phenomenon, in our cases to be transformed into a frog, a bee or a wolf/dog, can be interpreted in a psychopathological and diagnostic regard. Is it pathognomonic for a special disease? With the three case reports and a survey of the literature this paper deals with Lycanthropy trying to put this perhaps neglected topic back into the focus of psychiatric interest and place it in psychopathology. PMID- 11417262 TI - [The course of infantile autism through adulthood. An overview of long-term follow-up data]. AB - The symptoms of infantile autism were first described almost 60 years ago. In contrast to its course in puberty and adolescence, follow-up-data on the late course in adulthood are decidedly sparse. As the outcome of research in the literature, we found 21 methodologically heterogeneous follow-up-studies. The arithmetic mean age of all subjects investigated was 24.0 years. The results are supplemented by various case reports and sporadic biographical reports by affected persons. On the basis of the available data, the discontinuous and dynamic changes of course verified in puberty and adolescence are not applicable to the third and fourth decades to the same extent. Gains in competence and autonomy appear to develop in the vocational rather than in the domestic sphere. The significantly more favorable courses of the form described by Asperger are continued in adulthood. The disorder-associated lack of empathy and social interaction is by no means experienced in terms of self-satisfaction by those concerned but rather as a loss. Interpersonal sexual needs are expressed by a substantial proportion of autistic adults. The cumulative mortality rates of the follow-up-studies suggest that the mortality rate among autistic patients is higher than among their non-autistic peers. PMID- 11417263 TI - [Correlation of clinical and magnetic resonance imaging findings in patients with brainstem infarction]. AB - The aim of this study was the comparison of clinical and neurological findings in 30 patients presenting with ischemic brainstem lesions. These were localized in the midbrain in 4 cases, in the medulla in 12 cases and in the pons in 11 cases, while the remaining three patients demonstrated combined lesions. Symptoms were lesions of the pyramidal tract in 77% of cases, vertigo in 57% of cases, speech disturbances in 40% of cases and gait ataxia in 37% of cases. Cranial nerve lesions were evident in 87% of patients, while 33% of patients demonstrated a Horner syndrome. Brainstem lesions were diagnosed in 22 (73%) of patients on magnetic resonance imaging, while all 30 patients had clinical signs suggestive of brainstem lesions. We conclude that neuroradiological studies can provide helpful information regarding patients with brainstem lesions, but by no means replace exact neurological examination. PMID- 11417265 TI - [Peripheral nerve block: more questions than answers?]. PMID- 11417264 TI - [Geriatric Psychiatry Congress. Dusseldorf 12, 1-2, 2000]. PMID- 11417266 TI - [Peripheral nerve block. An overview of new developments in an old technique]. AB - General anaesthesia and peripheral neuronal blockade are techniques which were introduced into clinical practice at the same time. Although general anaesthesia was accepted significantly faster due to effective new drugs and apparent ease of handling, neuronal blockade has recently gained great importance. The reasons are in particular newer aids such as industrially produced catheter sets, nerve stimulators and ultrasound guidance which have facilitated that these economical techniques can be used not only for intraoperative anaesthesia but also for perioperative analgesia without any major risks for the patients. In parallel to epidural anaesthesia a change of paradigms has recently taken place using catheter instead of single-shot techniques. This allows the loading dose of the local anaesthetics to be installed in a safe way, to reload the dose when intraoperatively required and to extend the analgesia perioperatively by this technique using lower concentrations of the same drugs or drug combinations. A great number of short, middle or long acting local anaesthetics are available to choose the right drug for any particular case. Short and middle acting drugs are characterised by a faster onset compared to long acting drugs, but toxic plasma levels are seen during long time application causing seizures or drowsiness or by using prilocaine methemoglobin. Therefore long acting local anaesthetics such as bupivacaine, ropivacaine or levobupivacaine are the first choice drugs for long time application via peripheral nerve catheters for perioperative anaesthesia and analgesia. By using low concentrations of these potent drugs even for a longer period of time, no toxic plasma levels are seen with the exception of artificial intravasal injections. Additives such as opioids and alpha 2-sympathomimetics are also used. While the use of opioids is controversial, alpha 2-sympathomimetics are able to accelerate the onset and to extend the duration of regional anaesthesia and analgesia. PMID- 11417267 TI - [Gamma-hydroxybutyric acid-ethanolamide (LK 544). The suitability of LK 544 for sedation of patients in intensive care in comparison with midazolam]. AB - OBJECTIVE: In this phase 2 study gammahydroxybutyric acid-ethanolamide (GHB ethanolamide) was compared with midazolam for sedation of patients in the intensive care unit (ICU). GHB-ethanolamide is a new derivative of gammahydroxybutyric acid, a drug commonly used for sedation in intensive care patients. METHODS: A total of 29 non-intubated, spontaneously breathing patients following major surgery, were randomly assigned to 2 groups: group A (n = 14) received 150 mg/kg GHB-ethanolamide i.v. followed by 150 mg/kg/h. Group B (n = 15) received 0.025 mg/kg midazolam i.v. followed by 0.025 mg/kg/h. The degree of sedation was assessed over a 3-5 h period both clinically by the Ramsay Score and by the spectral frequency index (SFx), derived by continuous computerized EEG recording (CATEEM). RESULTS: EEG: the SFx showed a significantly deeper sedation compared to baseline values, 10, 60 and 120 min after start of sedation in the GHB-ethanolamide group. By contrast, no difference could be observed compared to baseline values in the midazolam group. A comparison between both groups showed a deeper sedation in the GHB group 60 and 120 min after start of sedation. The Ramsay Score increased from baseline values of 2.0 (2.0/2.0) to 3.0 (2.0/3.0) during sedation (Median (25th/75th percentile) and no significant differences could be observed between groups. In the Midazolam group the sedation of two patients had to be terminated because of side effects (Ramsay Score 6 and paradoxical, agitated reaction). CONCLUSIONS: GHB-ethanolamide produces adequate sedation for extubated and spontaneously breathing ICU patients. The drug might be safer than midazolam with regards to side effects such as respiratory depression. PMID- 11417268 TI - [Malignant neuroleptic syndrome after haloperidol administration]. AB - The neuroleptic malignant syndrome (NMS) is a rare complication of antipsychotic therapy. We report on a 65-year-old patient who was treated with haloperidol, diazepam and mirtazapin because of a severe depressive episode with psychotic symptoms. He exhibited most of the signs and symptoms characteristic of NMS, e.g.: hyperthermia, rigidity, elevated creatine phosphokinase, leukocytosis, elevated liver enzymes, reduced consciousness and autonomic nervous system disturbances. A secondary pneumonia was diagnosed 2 days after the onset of the NMS, which might have been due to chest wall rigidity. Intensive care treatment consisted of immediate discontinuation of the offending agent, supportive therapy with rehydratation and catecholamines as well as application of dantrolen. After 23 days of intensive therapy all pathological parameters were normalised and the patient was transferred to an internal ward. Three main theories on the pathogenesis of NMS exist: 1. blockade of central receptors, 2. a skeletal muscle target model and 3. sympathoadrenal hyperactivity. The differential diagnosis includes among others malignant hyperthermia and serotonin syndrome. PMID- 11417269 TI - [Interscalene plexus block. Anatomic requirements--anesthesiologic and operative aspects]. AB - The interscalene brachial plexus block with and without a catheter has become an indispensable method for anaesthesia and analgesia in shoulder surgery. Not only thorough knowledge of anatomy, but also accurate indication assessment and discussion with the surgeon regarding the location of access, is essential for the successful practice of this technique. Important and practical tips for implementation should especially help the less experienced, with special emphasis on correct positioning of the patient for surgery to avoid iatrogenic neural damage. Preoperative counselling of inevitable side-effects of the technique enhances the patient's acceptance and satisfaction. PMID- 11417270 TI - [The new international guidelines for cardipulmonary resuscitation: an analysis and comments on the most important changes]. AB - In August 2000, the American Heart Association and the European Resuscitation Council published the conclusions of the International Guidelines 2000 Conference on Cardiopulmonary Resuscitation and Emergency Cardiovascular Care which contains both the new recommendations and an in-depth review. The discussions and drafting began at a conference in March 1999, followed by a second conference in September 1999, both attended by approx. 250 participants and another conference in February 2000 which was attended by approx. 500 participants. Review of the current state of science, discussion and final consensus continued subsequently via email, conference calls, fax, and personal conversation. During the entire process, scientists and resuscitation councils from all over the world participated, with participants from the United States comprising approx. 60%, and scientists from outside of the United States comprising approx. 40%. In order to ensure that the CPR recomendations are not dominated by any given nation or resuscitation council, most topics were reviewed and interpretated by two scientists from the United States and two scientists from outside of the United States. Accordingly, changes in these new CPR recommendations are the result of an evidence-based review by worldwide experts. The most important changes in the recommendations according to the authors are discontinuation of the pulse-check for lay people, 500 ml instead of 800-1200 ml tidal volume during bag-valve-mask ventilation (FiO2 > 0.4) of a patient with an unprotected airway, verifying correct endotracheal intubation with capnography and an esophageal detector, employing mechanical devices such as interposed abdominal compression CPR, vest CPR, active-compression-decompression CPR, and the inspiratory threshold valve (ITV) CPR as alternatives or adjuncts to standard manual chest compressions, defibrillation with < 200 Joule biphasic instead of with 200-360 Joule monophasic impulses, vasopressin (40 units) and epinephrine (1 mg) as comparable drugs to treat patients with ventricular fibrillation, amiodarone (300 mg) for shock refractory ventricular fibrillation and intravenous lysis for patients who have suffered a stroke. PMID- 11417271 TI - [Changing culture--simulator-training as a method to improve patient safety. Report on an international meeting on medical simulation. Scottsdale: January 12 14, 2001]. PMID- 11417272 TI - [Is the use of tumescence anesthesia in general anesthesia contraindicated?]. PMID- 11417273 TI - [TIVA in unknown pregnancy]. PMID- 11417274 TI - [Pneumocephalus following spinal anesthesia]. PMID- 11417275 TI - [Preparation of laryngeal masks]. PMID- 11417276 TI - [Anesthesia for kidney transplantation]. PMID- 11417277 TI - [5-HT3 receptor antagonists for obstetric anesthesia]. PMID- 11417278 TI - [Physiological effects, pharmacology and indications for administration of magnesium]. PMID- 11417279 TI - Variable extension of the transitional zone in the medial structure of carotid artery tripod. AB - BACKGROUND: The goal of our study was to demonstrate the extension of the transitional zone (TZ) between elastic and muscular medial structure in carotid artery tripod. PATIENTS AND METHODS: A histologic study of 56 probes from 8 carotid artery tripods was performed. The probes were obtained from autopsies of 4 adults (mean age: 47.5 years, range 38 to 55 years) and were taken from 7 different topographic sites. RESULTS: At each level of the CCA (at 1 cm and 2 cm proximal to the bifurcation as well as at the bifurcation) we observed an elastic arterial type in 24 (42.8%) probes with 11 to 20 (medium 15.0) elastic fibers per view field (200 x magnification) in the media. In contrast the histologic structure of the ICA and ECA varied as follows: in 8 sections (14.3%) elastic arterial type with 11 to 16 (medium 13.1) elastic fibers, in 11 sections (19.6%) muscular arterial type with 2 to 5 (medium 3.5) elastic fibers and in 9 sections (16.1%) a transitional arterial type with 6 to 8 (medium 6.7) elastic fibers in the media. Atherosclerotic lesions have prevented the assessment of the arterial type in 4 probes (7.1%). The TZ in the medial structure of carotid artery tripods is exclusively localized in the ICA/ECA but not in the CCA. The ICA/ECA presented a TZ with a length up to 0.5 cm (4 probes; 25%), up to 1.5 cm (4 probes; 25%) and longer than 1.5 cm (6 probes; 37.5%). CONCLUSIONS: In this study we confirmed that in the carotid artery tripod, a TZ--an arterial segment with transition from elastic to muscular type--does exist, involving a variable length. Furthermore studies on the impact of the biomechanical properties of the TZ as a potential factor in atherosclerotic disease are justified. In addition, the complex biomechanical behavior of the TZ should be considered prior to interventional procedures. PMID- 11417280 TI - Multicentre, randomised controlled trial of four-layer bandaging versus short stretch bandaging in the treatment of venous leg ulcers. AB - BACKGROUND: Aim of the study was to compare the healing rates of venous ulcers obtained with four-layer bandages (4LB) versus short stretch bandages (SSB). DESIGN: Multicentre, randomised controlled trial performed in 5 centres of the Netherlands and in 2 centres in Austria ("PADS-study" = Profore Austrian Dutch Study). PATIENTS AND METHODS: 112 patients (53 treated with 4LB and 59 treated with SSB) completed at least one post-treatment follow-up, 90 completed the study. Bandaging and ulcer assessment was performed at weekly intervals. Randomisation was carried out for each centre and was stratified according to the size (more or less than 10 cm2) of the ulcerated area. Local therapy consisted of plain absorbing, non-adherent dressings. Time to complete healing was recorded up to a maximum of 16 weeks. The two treatment-groups were comparable regarding their baseline-characteristics. RESULTS: In total 33/53 (62%) of ulcer-patients were healed in the 4LB group, compared with 43/59 (73%) in the SSB group (difference 11%, 95% CI -28% to 7%). 77% of the ulcers with an initial area less than 5 cm2 healed as compared with 33% of the larger ulcers. The different healing rates in the centres could be explained by the different sizes of the treated ulcers. Based on Kaplan-Meier estimates the median healing time was 57 days for the 4LB (95% CI 47-85 days) and 63 days for the SSB (95% CI 43-70 days). CONCLUSION: The ulcer healing rate and the median healing time did not differ among the two types of bandages. The main discriminant criterion for healing was the initial ulcer size. In centres who are experienced users of short-stretch bandages, no statistically significant different healing rates of venous ulcers could be found after 4LB or SSB. PMID- 11417281 TI - Urgent carotid endarterectomy for stroke in evolution. AB - BACKGROUND: Carotid endarterectomy (CEA) is well established as the elective treatment for moderate or severe carotid stenoses with a history of neurologic symptoms. In contrast, the merits of carotid revascularisation performed in emergency in patients with acute stroke or fluctuating neurological deficit remain controversial. PATIENTS AND METHODS: A total of 445 CEAs were performed on 424 patients for 212 (48%) asymptomatic and 233 (52%) symptomatic carotid stenoses within a 5 years period between January, 1995, and December, 1999. Of the latter, CEA was performed in emergency on 16 patients (3.8%) within 4 to 24 hours after the onset of symptoms. Patients selected for urgent surgery fulfilled the following criteria: acute onset of fluctuating hemispheric neurological symptoms, significant carotid pathology, absence of cerebral hemorrhage, uncompromised vigilance and stable cardiopulmonary conditions. Selected patients presented with a crescendo-TIA (n = 7) or fluctuating neurological deficits (n = 9) corresponding to a contralateral carotid stenosis. RESULTS: Following CEA, the neurological deficits improved instantaneously to complete recovery in 9 patients. The symptoms of 4 patients improved to non-disabling deficits, remained unchanged in one and worsened in 2 patients from hemihypaesthesia to hemiparesis. 14/16 patients were discharged within 8 days after admission. The neurologic status after discharge did not deteriorate in any of the patients during follow up of 19.3 +/- 13 months, but improved in 4 of the patients. CONCLUSION: Our retrospective study suggests that rescue CEA may be beneficial for selected patients with stroke in evolution and fluctuating neurological deficits. Careful adherence to selection criteria, intraoperative shunting, intensive care post surgery surveillance and an experienced team are recommended. PMID- 11417282 TI - Glove perforation rate in vascular surgery--a comparison between single and double gloving. AB - BACKGROUND: In surgery intact gloves act as a sterile barrier between surgeon and patient. The impermeable gloves protect the surgeon from bloodborne pathogens such as HIV, hepatitis B, and hepatitis C. On the other hand, the surgical wound is protected from micro-organisms from the skin of the surgeon. One objective of this study was to compare puncture rates between the double gloving color indication system and single-use gloves and the other to determine the extent to which glove perforations remain undetected during the course of vascular surgical operations. PATIENTS AND METHODS: The study material comprised all gloves used in vascular surgical operations at Satakunta Central Hospital for a period of two months. The analysis was made by the glove type in a prospective and randomised manner. Gloves were tested immediately after the surgical procedure using the approved standardized water-leak method. With this method the glove is filled with water using a special filling tube, and the water-filled glove is then checked for two minutes to detect any holes. The gloves used in this study were either double gloves with indicator, or the standard glove used at our hospital. RESULTS: In 73 operations altogether 200 gloves were tested, half of them were double gloves and half were single gloves. The perforation occurred in the double gloves 3 times and with single gloves 12 times. The overall perforation rate was 15 out of 200 gloves (7.5%). The detection of perforation during surgery was 60%. Most frequently the perforation was located in the second finger of the left hand, 9 out of 15 perforations. CONCLUSION: In view of the critical importance of safety at work both transmitting the pathogens from the skin of the surgeon to the wound and transmitting the bloodborne pathogens from the patient to the surgeon, it is very important to use double gloving at least in operations where there is a high risk of glove perforation. PMID- 11417283 TI - [Inguinal hernia as a diagnostic error in ruptured groin aneurysm]. AB - Anastomotic aneurysms observed with an incidence of 0.5% to 5.0% are considered a known complication following arterial surgery, especially when fabric grafts in the inguinal region are implanted. An anecdotal report is presented describing a 64-year old male patient, who developed, 10 years following an autologous femoro tibial vein graft, a huge mass in the left groin. The lesion was considered an incarcerated inguinal hernia and the patient was admitted to the Department of Surgery for emergency repair. Clinical examination, duplexsonography and CT scan clarified the diagnosis of an aneurysm with a diameter of 13 cm. The aneurysm was resected, and a femoro-profundal vein graft was implanted orthotopically, the graft was covered with a sartorius muscle flap. The postoperative course was uneventful. The diagnosis is suspected by clinical examination and usually confirmed by duplexsonography. The exact etiology of suture line aneurysms is unknown; in the present case progression of the underlying arteriosclerotic arterial disease after a follow up of 10 years is likely. For the treatment the usual methods of complicated aneurysm repair and preservation of the arterial circulation--using autologous in situ methods or extraanatomic bypass grafts- with additional biologic coverage are at hand. PMID- 11417284 TI - [Purulent arthritis of the knee joint and septic microembolism of the foot as postoperative complications of an infected femoral anastomosis aneurysm]. AB - Septic peripheral embolism due to infective complication of vascular infrainguinal procedures has been occasionally reported. The combination, however, of purulent arthritis of the knee joint and septic microembolism of the foot, as consequence of an infected pseudoaneurysm, is a very rare entity. We report a case of an infected anastomotic femoral aneurysm, following an iliofemoral "in situ repair" which was complicated with purulent arthritis of the knee joint and multiple small necrotic lesions of the plantar surface of the foot. Treatment included removal of the infected graft, an extra-anatomic revascularization and arthrotomy with pus evacuation. The patient had an uneventful postoperative course and remained well at four-year follow-up, with a mild dysfunction in the knee joint flexion. PMID- 11417285 TI - [Hypothenar-hammer syndrome]. AB - The unilateral distal arterial lesions still give problems in classification and differential diagnosis specially in younger patients. We report on a 45 years old male patient with an aneurysm of the distal ulnar artery and superficial palmar arch. The chronic occupational trauma of the artery led to the clinical picture of hypothenar hammer syndrome. PMID- 11417286 TI - Surgical management of retro-aortic left renal vein in combined abdominal aortic and coronary surgery. AB - Although generally retro-aortic left renal vein is a rare anatomic finding, it occurs in 0.8% of the patients admitted for abdominal aortic aneurysm surgery. Surgeons fear fatal bleeding during clamping of the aorta, caused by a more caudal insertion of the retro-aortic left renal vein and a greater vulnerability of the anomalous tissue. Once such a complication occurs, a reconstruction of the retro-aortic left renal vein using a synthetic graft should be performed to obtain adequate renal venous flow and maintain renal function. PMID- 11417287 TI - Vascular complications after the treatment with Ilizarov external fixators. AB - Iatrogenic vascular injuries from external fixation in orthopaedics and traumatology are frequent. Three cases of vascular injuries after the treatment with Ilizarov external fixators were treated at our institution. These include two cases of pseudoaneurysms and one case of acute ischaemia of the lower limb. Two patients became symptomatic only after removal of the fixator. In all cases, the diagnosis was made by color flow duplex sonography. All vascular injuries needed surgical repair. PMID- 11417288 TI - [Thromboembolism]. PMID- 11417289 TI - [Presidential address at the 32nd Annual Congress of the Austrian Society of Vascular Surgery, Vienna, 27-30 September 2000]. PMID- 11417290 TI - [Clinical evidence]. PMID- 11417291 TI - Long-term oral anticoagulation therapy of peripheral arterial disease. AB - This review article summarizes clinical data on adjuvant long-term oral anticoagulation therapy (OAC) of peripheral arterial disease (PAD). It analyzes the underlying risk model of oral anticoagulation. Definitions of runoff patterns, of major and minor bleeding complications, of predictors of major bleedings as well as a classification of patient risk groups are described. The indication for OAC treatment of chronic limb ischemia remains still due to an individual decision. Clinical decision making is facilitated by the risk model. Improved oral anticoagulation control results in fewer bleeding complications. Studies on patient weekly self-testing and self-dosing which support this hypothesis are reviewed in the context of adjuvant OAC therapy. PMID- 11417292 TI - [Cystic adventitial degeneration. An important differential diagnosis in intermittent claudication]. AB - The cystic adventitial degeneration is a rare disease, but an important differential diagnosis in patients with intermittent claudication. The amount of fluid in the cysts may vary and cause an intermittent compression of the artery. This explains the frequently intermittent symptomatology, leading to a critical ischemia if there is a complete obstruction of the artery. The histologic findings indicate that adventitial cysts are true ganglions. The intramural, uni- or multilocular cysts contain a gelatinous, muciform fluid. The diagnosis of a cystic adventitial degeneration should be considered in cases of isolated stenosis or occlusion of the popliteal artery. Realtime ultrasound helps to establish the diagnosis. The standard treatment has been surgical and has consisted a resectional and non-resectional technique. The ultrasound-directed percutaneous aspiration as a less invasive technique seems to be an effective treatment for this condition. The course, diagnostic and therapy of four patients is demonstrated. PMID- 11417293 TI - Flow associated (endothelium dependent) vasodilation and TSH-levels in young normotensive and normoglycemic subjects. AB - BACKGROUND: Endothelial dysfunction (ED) is regarded as an early step in the development of atherosclerosis. Recent experimental data showed a crosstalk between endothelial NO-synthase activity and thyrotropin production. Therefore we studied whether basal TSH can predict flow associated vasodilation (FAD) in a cohort of healthy young subjects with normal TSH levels. PATIENTS AND METHODS: FAD was evaluated in 60 normotensive and normoglycemic subjects (mean age 34 years; range 18-50). The mean thyrotropin level was 1.43 +/- 0.11 microU/ml (range 0.18-3.52 microU/ml). RESULTS: Comparing subjects in the upper, middle and lower tertile of TSH (2.38 +/- 0.14 microU/ml, 1.23 +/- 0.04 microU/ml and 0.65 +/- 0.06 microU/ml respectively) there was no difference in terms of the classical cardiovascular risk factor profiles (24 h blood pressure, HDL- and LDL cholesterol, triglycerides, oral glucose load and body fat content). Regarding the vascular parameters, we could neither find an independent association with FAD (7.0 +/- 1.1%, 6.4 +/- 1.0% and 5.8 +/- 1.1% respectively) nor with endothelial independent vasodilation (after application of glycerol trinitrate GTN, 17.3 +/- 1.9%, 18.4 +/- 1.7% bzw. 17.5 +/- 1.6% respectively) between the groups. Furthermore, we could not find a significant association between free thyroid hormones (fT3/fT4) and FAD or GTN-induced vasodilation. CONCLUSION: TSH has no predictive value towards endothelial dysfunction in subjects with thyrotropin levels within the normal range. PMID- 11417294 TI - Psychological and physical reactions on children after the Hanshin-Awaji earthquake disaster. AB - Children who experienced the Hanshin-Awaji Earthquake Disaster were followed to ascertain how the psychological and physical reactions after this disaster changed. Changes observed in the symptoms of children at one and two years after the earthquake were compared between those who had lived in severely damaged area (level 7 on the Japan Meteorological Agency intensity scale) and those who had lived in mildly damaged area (less than 5 on the same scale). The survey was conducted using a questionnaire filled out by the children's parents. Two years after the earthquake, the children had returned to normal in terms of their physical conditions, even in the severely damaged area. However, symptoms of PTSD (Post-Traumatic Stress Disorder) such as persistent reexperiencing, persistent avoidance, and increased arousal were significantly more frequently found among children from the severely damaged area than among those from the mildly damaged area. To evaluate the psychological and physical reactions after the disaster is very important in order to support the children when large-scale disasters occur. PMID- 11417295 TI - Splenic abscess, pleural effusion and severe anemia caused by Salmonella typhi. AB - Various complications related to gastrointestinal system, central nervous system, and skeletal system may be observed during typhoid fever, but splenic abscess and pleural effusion are rarely encountered. A 12-year-old boy was admitted with fever, fatigue and pallor. On examination he had hepatosplenomegaly and severe anemia. He was diagnosed as having Salmonella typhi infection complicating with splenic abscess, pleural effusion and severe anemia, and successfully treated with percutaneous drainage with ultrasonography and antibiotics. In conclusion we would like to emphasize that typhoid fever should also be considered in patients with hepatosplenomegaly and severe anemia, and percutaneous drainage with ultrasonography may successfully be used in management of splenic abscess in typhoid fever. PMID- 11417296 TI - Evaluation of speech function after mesopharyngeal reconstruction with radial forearm flap. AB - This study examined postoperative speech function in ten patients who underwent mesopharyngeal resection and reconstruction using radial forearm flap. Patient age ranged from 49 to 71 years (mean 60 years) with 9 males and 1 female. Speech function was assessed by articulatory and velopharyngeal function. Articulatory function was judged by intelligibility of monosyllables and sentences. The percentage of 100 Japanese monosyllables perceived as correct in each case were scored. Intelligibility of sentences was evaluated using the Hirose standard. Velopharyngeal function was fiberoptically examined both in soft-blowing and at rest, and visual information about velopharyngeal closure was obtained. Correlation between the extent of resection and the speech function was then analysed. In order to maintain over 50% speech intelligibility and an excellent state in the Hirose standard, the extent of resection in the soft palate is particularly important. This result has to be explained in correlation with the resected range of the levator muscle, which is important for velopharyngeal closure. Surgically treated mesopharyngeal cancer patients face several functional problems. In order to counsel patients more meaningfully before surgical intervention, information about the postoperative speech function should be made available. The goal of reconstruction is to achieve acceptable speech in order to maintain the patients' quality of life. The results in this study reveal that resection which doesn't extend to the opposit side of the soft palate, can achieve the goal by utilization of the foream flap, because its pliable nature brings out the residual function of the levator. PMID- 11417297 TI - Immunohistochemical study of P53, PCNA and AFP in hepatocellular carcinoma, a comparison between Indonesian and Japanese cases. AB - Mutations of p53 as a tumor suppressor gene in hepatocellular carcinoma (HCC) have been reported to occur with varying frequency in different geographic regions, which might be different etiology for HCC. Overexpressions of p53 (well known for its implications in mutations of the p53 gene), PCNA and alpha fetoprotein (AFP) have been reported to be associated with carcinogenesis and/or tumor progression and poor prognosis in various types of cancer. To estimate the geographical difference of the p53 gene, PCNA and AFP in HCC, we examined 14 Japanese HCC cases, 8 Indonesian HCC cases, and 27 Indonesian chronic active hepatitis (CAH) or liver cirrhosis cases, using immunohistochemical approaches. Overexpression of p53 was identified in 37.5% of Japanese HCC, in 62.5% of Indonesian HCC and none in CAH. The mean PCNA Labeling Index of Japanese HCC, Indonesian HCC and CAH was detected in 48.6%, 30.4%, and 43.5%, respectively. AFP was detected in 35.7% of Japanese and 25% of Indonesian HCC. The rate of p53 overexpression in Indonesian HCC was as high as in HCC of southern part of China, which might share the similar etiology in both regions. PMID- 11417298 TI - Characteristics of HIV infection/AIDS in Korea. PMID- 11417299 TI - Acid secretion from a heterotopic gastric mucosa in the upper esophagus demonstrated by dual probe 24-hour ambulatory pH monitoring. AB - Heterotopic gastric mucosa in the upper esophagus is frequently found during endoscopic examination. Although most patients with heterotopic gastric mucosa of the upper esophagus, referred as inlet patch, are asymptomatic, symptomatic patients with complications resulting from this ectopic mucosa have also been reported. Acid secretion by the inlet patch has been suggested in some reports. We report a case of heterotopic gastric mucosa in the upper esophagus, with secretion of acid, demonstrated by continuous ambulatory pH monitoring, and the improvement of pharyngeal symptoms after the use of a proton pump inhibitor. PMID- 11417300 TI - Association of chronic hepatitis C virus infection and diabetes mellitus in Korean patients. AB - BACKGROUND: It has been suggested that chronic hepatitis C virus (HCV) infection is associated with diabetes. The aim of this study was to establish a potential relationship between chronic HCV infection and diabetes mellitus in Korean patients. METHODS: We performed a prospective analysis of 404 patients with chronic viral hepatitis or liver cirrhosis who visited our hospital and analyzed whether age, sex, body mass index, alcohol consumption, hepatitis B virus (HBV) infection, HCV infection and cirrhosis were associated with diabetes. We also enrolled 627 diabetic patients and the seroprevalence of HBV surface antigen (HBsAg) and anti-HCV was determined. RESULTS: Diabetes was observed more frequently in individuals with HCV infected chronic liver disease (24.0%) than in those with HBV infected (10.4%) (p < 0.05). Univariate analyses revealed that age, alcohol consumption and HCV infection were significant independent predictors for diabetes. The mean age of the patients with HCV infected chronic liver disease was higher than that of HBV infected (56 +/- 16 vs 44 +/- 13, p < 0.05). The prevalence of diabetes in HCV infected group was higher than that in HBV infected group in the age of 41-60 (p < 0.05). In diabetic group, the seroprevalence of HBsAg positivity was 4.5% and that of anti-HCV was 2.1%. CONCLUSION: Our study demonstrates an association between diabetes and chronic HCV infection in Korean patients. The prevalence of diabetes in patients with HCV infected chronic liver disease is higher than that in those with HBV infected. Age and alcohol consumption are another risk factor for diabetes in patients with chronic viral liver disease. PMID- 11417301 TI - Clinical significance of anti-endothelial cell antibody in renal transplant recipients. AB - In order to evaluate the role of anti-endothelial cell antibody (AECA) in acute rejection in renal transplantation, serum AECA IgG titers were measured in 68 healthy controls, 111 chronic hemodialysis (HD) patients and 58 first renal transplant recipients. The AECA titer in hemodialysis patients was higher than in healthy controls (13.9 +/- 5.0 vs. 4.8 +/- 2.3 U/mL, p < 0.01). In transplant recipients, AECA titers were not affected by dialysis mode (HD vs. CAPD vs. non dialysis; 9.6 +/- 7.6 vs. 7.9 +/- 3.9 vs. 11.9 +/- 3.1 U/mL, p > 0.05). After renal transplantation, AECA titer was decreased significantly (vs. 4.7 +/- 3.6 U/mL. p < 0.01). The serum AECA IgG titers increased significantly in recipients with acute rejection (6.9 +/- 3.1 vs. 13.5 +/- 9.9 U/mL, p < 0.01), but decreased to 5.6 +/- 3.0 U/mL (p < 0.01) after formal rejection therapy. In the recipients with acute rejection (n = 27), the pre-renal transplant AECA titer was higher than in that without acute rejection (14.0 +/- 8.6 vs. 7.7 +/- 3.8 U/mL, p < 0.01). The results of this study lead us to conclude that pre- and post-renal transplant AECA titer might be a useful predictor for acute rejection and useful for monitoring acute rejection in renal transplant recipients. PMID- 11417302 TI - Detection of malignant cells in pleural fluid or ascites by CD44v8-10/CD44v10 competitive RT-PCR. AB - BACKGROUND: CD44 is a cell surface adhesion molecule which has been implicated in various biologic functions as lymphocyte homing and activation, cellular migration and extracellular matrix adhesion. Over-expression of CD44v8-10 has been found in several cancers and is considered to be associated with tumor progression and metastasis. Recently, a novel molecular method, CD44v8-10/CD44v10 competitive reverse transcription-polymerase chain reaction(RT-PCR) has been developed for detecting cancer cells over-expressing CD44v8-10. METHODS: We analyzed from benign and malignant pleural effusion and ascites by CD44 competitive RT-PCR and compared to the conventional cytology. RESULTS: The CD44 competitive RT-PCR analysis showed that all the 24 samples associated with benign disease presented a predominant expression of the CD44v10 transcript (v8-10/v10 ratio: 0.126-0.948), whereas 6 of 7 malignant pleural samples associated with cytology positive cancer expressed the CD44v8-10 transcript (v8-10/v10 ratio > 1.00). CONCLUSION: These results indicate that CD44 competitive RT-PCR assay is a useful and adjunct to cytological examination in cancer diagnosis, especially in detecting exfoliated cancer cells in pleural effusion. PMID- 11417303 TI - A case of BOOP developed during bucillamine treatment for rheumatoid. AB - We describe a patient with rheumatoid arthritis(RA) who developed bronchiolitis obliterans organizing pneumonia(BOOP) during the treatment of bucillamine. A 51 year-old man was admitted to the hospital for an abnormal shadow on his chest radiograph. He had been diagnosed as having RA 3 years previously and had been receiving 200 mg of bucillamine for 21 months. Two months prior to admission, he presented with a cough and his chest X-ray showed opacities in both lower lungs. He was treated with antibiotics for 2 months after the development of cough and lesions on the chest X-ray, but the symptoms and lung lesions became more aggravated. On admission, an HRCT revealed airspace consolidations in the subpleural space of both basal lungs and a CT-guided fine needle aspiration biopsy showed Masson's body filling air space, interstitial infiltration of acute and chronic inflammatory cells and type II cell hyperplasia, consistent with BOOP. Bucillamine was stopped and 50 mg of prednisolone was administered. His symptoms and infiltrations on the chest X-ray resolved. We suggest that bucillamine should be considered as a drug possibly associated with BOOP. PMID- 11417304 TI - A case of malignant proliferating trichilemmoma of the scalp with multiple metastases. AB - We report a case of malignant proliferating trichilemmal tumor showing multiple distant metastases. The patient demonstrated a round mass in the right occipital area for 12 months and the lesion grew rapidly to assume 8 x 6.5 x 4 cm in diameter, with areas of superficial erosion and crusting within the recent 3 months. The entire lesion was removed with a wide surgical excision. It recurred on the neck area 4 months after excision and the lesion was removed with surgical resection again. There was evidence of multiple metastases on CNS and mediastinal lymph nodes after 6 months. The patient was treated with cisplatin and etoposide combination chemotherapy and a partial response was achieved. PMID- 11417305 TI - A case of hypothermia combined with systemic lupus erythematosus. AB - Systemic lupus erythematosus(SLE) is a prototypic autoimmune disease affecting various organ systems. Hypothermia is a rare manifestation of SLE. We experienced a case of SLE combined with hypothermia. A 36-year-old woman, who had been diagnosed as SLE 3 days before admission, admitted complaining of mental confusion. After admission, her body temperature, initially 36.1 degrees C, became 32.6 degrees C. Her core body temperature was less than 35.0 degrees C. Despite of warming with heating lamp and blankets, her core temperature did not reach 35.0 degrees C during 18 hours. Ten days later, her temperature exceeded 36.0 degrees C. PMID- 11417306 TI - A case of nodular scleritis in association with Behcet's disease. AB - Approximately 50% of patients with scleritis are known to be associated with systemic connective tissue diseases or vasculitic diseases such as rheumatoid arthritis (RA), Wegener's granulomatosis, relapsing polychondritis, and systemic lupus erythematosus. The patients with scleritis in association with Behcet's disease (BD) have been rarely described in the literature. We report a 46-year old female patient with BD who developed the nodular scleritis. PMID- 11417307 TI - Relevance of vacA genotypes of Helicobacter pylori to cagA status and its clinical outcome. AB - BACKGROUND: Determination of vacA mosaicism may be important because specific Helicobacter pylori vacA genotype can be used to predict different clinical outcome. The aim of this study was to assess the relationship of vacA genotypes of Helicobacter pylori to cagA status and its development of peptic ulcer diseases in Korean patients. METHODS: Gastric biopsy specimens were obtained from 53 patients with gastric ulcer(GU), 57 with duodenal ulcer (DU) and 26 with chronic gastritis(CG) patients; all patients were infected with Helicobacter pylori. Bacterial mRNAs in the gastric mucosa were amplified by RT-PCR, using synthetic oligonucleotide primers specific for the vacA and the cagA gene. Patients with vacA s1 subtype were further examined to determine whether they had s1a or s1b subtype. RESULTS: There was no correlation in frequency of vacA s1 and/or s1a genotype between CG and either GU or DU, as the vacA s1 and s1a/m1 were present in the majority of strains independent of clinical status(s1; 100.0% versus 94.3% or 93.0% and s1a/m1; 76.9% versus 62.3% or 64.9%, respectively). Likewise, there was no difference in the prevalence of the cagA gene between CG and either GU or DU patients (92.3% versus 90.6% or 98.2%, respectively). In addition, the cagA-negative status did not predict the presence of vacA s2 genotype. CONCLUSION: These results strongly suggest that either cagA or vacA s1 and/or s1a is not proved to be a useful marker to distinguish disease-specific Helicobacter pylori strains for the development of peptic ulcer diseases in Korean patients. PMID- 11417308 TI - Transesophageal echocardiography in the assessment of patients presenting with ischemic cerebral events without previous evidence of a cardiac source of emboli. AB - Transesophageal echocardiography (TEE) is accepted as a valuable tool in the evaluation of ischemic stroke patients, particularly in the young and in cases of unknown cause. However, the real clinical impact of additional TEE data remains to be defined. PURPOSE: The aim of this study was to present our experience with TEE in ischemic stroke patients without previous evidence of a cardiac source of emboli. METHODS: From March 1991 to June 2000 we studied 172 patients (80 males, 92 females, mean age 43 +/- 12 years presenting with a transient ischemic attack or a recent cerebral infarction who had no previous evidence by clinical assessment, electrocardiogram and transthoracic echocardiography of a cardiac of emboli source. The population was divided into two groups: Group A--age < or = 45 years, n = 101 (43 males, 58 females, mean age 34 +/- 7 years) and Group B--age > 45 years, n = 71 (36 males, 35 females, mean age 54 +/- 7 years). Information was gathered from clinical records. In every case TEE included Doppler color flow imaging and multiple contrast injections (agitated saline) with and without Valsalva maneuver. RESULTS: A potential cardiac of emboli source was found in 29% of the patients, with a higher prevalence in the older group (group A--25%, Group B--35%, ns). Atrial septal abnormalities accounted for most of the detected findings (23 patent foramen ovale, 17 atrial septal aneurysms and two atrial septal defects, representing altogether 71% of the findings). The other detected anomalies were distributed as follows: aortic plaques--six, mitral valve prolapse -five vegetations--four, thrombus in left atrial appendage--two. Only six patients (3.5%) had abnormalities which in themselves determined a specific approach, which were found mostly in the older group (Group A--two vegetations; Group B--two vegetations, two thrombi). CONCLUSIONS: TEE identified additional findings with possible embolic potential in a considerable number of cases, the majority of which were of a debatable cause-effect relation. A higher diagnostic yield and clinical relevance was found in older patients, which argues against age being used as a selective criterion for it. Our experience suggests that routine TEE in this setting is of questionable value and has little impact on clinical management, for which reason TEE referral should be decided on an individual patient basis. PMID- 11417309 TI - Early and medium term results of tailored therapy for heart failure. AB - Patients (pts) with advanced chronic heart failure, in NYHA functional class IV, refractory to conventional medical therapy, show a poor short-term survival prognosis. Heart transplant remains the therapy of choice but it can currently be performed in only a minority of pts. Therapy tailored to hemodynamic goals has been suggested as a potential alternative for patients with advanced congestive heart failure. Intravenous and, subsequently, oral vasodilators (v) and diuretics (d) are titrated, in order to achieve specific hemodynamic parameters: systolic arterial pressure (SAP) > 80 mmHg, pulmonary wedge pressure (PWP) < 15 mmHg, right atrial pressure (RAP) < 8 mmHg, and systemic vascular resistance (SVR) < 1200 dynes.sec.cm-5. AIM: To assess short and medium term (two years) results of a tailored therapy management program for treatment of patients with advanced heart failure. METHODS: 27 pts (19 males, 61 +/- 10 years), NYHA functional class IV, with dilated cardiomyopathy (13 idiopathic, 10 ischemic, 4 hypertensive), 17 with exclusion criteria for heart transplantation, were included. Echocardiographic left ventricular end-diastolic dimension and ejection fraction were 68 +/- 8 mm and 20 +/- 9%, respectively. Initial (i) serum sodium (Na+) was 136 +/- 5 mEq/l and i serum creatinine (Cr) was 1.4 +/- 0.8 mg/dl. Baseline, at referral, hemodynamics: SAP = 125 +/- 23, PWP = 23 +/- 6, RAP = 12 +/- 6, cardiac index (CI) = 1.9 +/- 0.5 l/min/m2, SVR = 2193 +/- 670. Using bedside right heart catheterization (Swan-Ganz catheter) we set out to achieve the above hemodynamic goals. RESULTS: 1) v and d used: sodium nitroprusside--cumulative dose = 196 +/- 121 mg, captopril--daily dose (dd) = 157 +/- 95 mg, isosorbide dinitrate--dd = 91 +/- 57 mg, hydralazine--dd = 95 +/- 67 mg, and furosemide--dd = 105 +/- 70 mg; 2) final (f) hemodynamic parameters on tailored therapy: SAP = 109 +/- 20*, PWP = 12 +/- 3*, RAP = 4 +/- 3*, CI = 2.5 +/- 0.6*, SVR = 1317 +/- 340* (*: p < 0.001 vs baseline); 3) duration of invasive monitoring was 3.0 +/- 1.9 days; 4) f Na+ = 134 +/- 5, and f Cr = 1.5 +/- 0.8 (NS vs i); 5) there was one (4%) in-hospital death; functional class of discharged pts: III--4 pts, II--18 pts, I--4 pts; 6) nine pts (35%) died after discharge--three due to refractory heart failure and six (including two potential heart transplant candidates) had sudden death; 7) actuarial survival (Kaplan-Meyer method): at 6 months (m)--80%, 12 m--71%, 18 m- 64%, 24 m--55%; 8) after a mean follow-up of 18 +/- 8 m, functional class in survivors was: NYHA IV--2 pts, III--4 pts, II--8 pts, and I--3 pts. CONCLUSIONS: Therapy tailored to hemodynamic goals is a valid approach for pts with advanced heart failure, showing good hemodynamic and functional short-term results, and reasonable survival at two years. The significant incidence of sudden death demands strategies for risk stratification and a search for prophylactic measures in this population. PMID- 11417310 TI - Severe heart failure: role of tailored therapy. PMID- 11417311 TI - [Mechanisms underlying endothelin-1 effects on myocardial function]. AB - INTRODUCTION: Endothelin-1 (ET-1) has potent vasoconstrictor, growth promoting and positive inotropic properties. Its effects on the intrinsic properties of the myocardium were recently described. The present study investigated the mechanisms underlying those effects. METHODS: The myocardial effects of 1 and 10 nM of ET-1 were evaluated in isolated rabbit papillary muscles (n = 9) and human atrial trabecula from CABG patients (Krebs-Ringer; 1.8 mM CaCl2; 35 degrees C). In papillary muscles the effects of 1 nM ET-1 were also studied in the presence of: (i) a selective ETA receptor antagonist, BQ-123 (0.1 microM; n = 9); (ii) a selective ETB receptor antagonist, BQ-788 (0.1 microM; n = 6); and (iii) an Na+/H+ exchanger inhibitor, methyl-isobutyl-amiloride (MIA; 1 microM; n = 6). Only significant results (mean +/- SE, p < 0.05) are given, expressed as delta % baseline. RESULTS: In AT by papillary muscles, 1 nM of ET-1 increased 64 +/- 16%, dT/dtmin 39 +/- 13% and decreased PT by 11 +/- 2%. The analysis of atrial strip contractions yielded similar results. In papillary muscles the effects of ET-1 were not affected by BQ-788, yet they were abolished by BQ-123, and reduced by 44% by MIA. CONCLUSIONS: The action of ET-1 on myocardial function is similar in human and non-human myocardium. The myocardial effects observed in the present study are mediated by the binding to ETA receptors, and partially dependent on Na+/H+ exchanger activation. PMID- 11417312 TI - [Assessment of aortic atherosclerosis with transesophageal echocardiography]. AB - OBJECTIVE: To image the thoracic aorta by transesophageal echocardiography (TEE) and study atherosis (morphology and extent of atheroma plaques) and sclerosis (stiffness) and secondarily correlate them with serum lipid levels (cholesterol, HDL, LDL and triglycerides). METHODS: We studied 29 patients (pts) who underwent TEE electively (male 18 pts, age 57.8 +/- 14.6 years). The parameters evaluated were: the stiffness coefficient = ln (PAsist/PAdiast)/(Dsyst/Dmin), and the morphology, location and extent of atheroma plaques. The systolic distension (Dsyst) was the difference between the maximal and the minimal dimensions (Dmin) of the aortic diameter measured by M mode. The lesions were classified in 4 degrees (0-3): 0--normal intima, 1--intimal thickening, 2--atheroma, 3- complicated lesion. Five aortic segments were studied: arch, D1-D4 (descending aorta at 5 cm intervals from the first 25 cm distal of the incisors line). We calculated the individual score = 1 x theta 1/180 + 2 x theta 2/180 + 3 x theta 3/180, theta n represents the angles occupied by the lesions and n (1-3) the severity of atherosis of each lesion. The total atherosis index (TAI) was sigma scores/(n. degree of visualized segments). RESULTS: The arch was not visualized in 3 pts, and the segment D4 was only visualized in 3 pts. TAI mean = 0.82 +/- 0.74, stiffness coefficient mean (SC) = 9.56 +/- 15.072. There were no significant correlations between the lipid levels and the TAI or SC. The only significant positive correlations were: TAI vs age (r = 0.62, p < 0.001) and SC and diastolic blood pressure (BP) (r = 0.42, p < 0.05). CONCLUSIONS: The best visualized segments belong to the descending aorta (25 to 40 cm from the incisors). In this group of patients the lipid levels did not seem to be a preponderant factor in aortic atherosclerosis. The most important factors were age for atherosis and BP for sclerosis. PMID- 11417313 TI - [Complete congenital atrioventricular block]. AB - Congenital atrioventricular block is defined (Yater) as a documented conduction defect in a young subject with unquestionable bradycardia in the absence of a history of infection which might have caused the condition. This condition may be isolated or associated with structural cardiac disease. Isolated block is often observed in patients with mothers suffering from autoimmune disease, often clinically latent. Anti-Ro/SS-A and La/SS-B antibodies cross into the fetal circulation and cause inflammation of the fetal conduction system. The diagnosis of the conduction defect is sometimes made during fetal life by echocardiography. Echocardiography assesses fetal hemodynamic status and may detect signs of fetal deterioration. Long-term follow-up studies have revised the previously considered good prognosis of isolated congenital atrioventricular block but advances in cardiac pacing provide satisfactory treatment. Follow-up by Holter monitoring, exercise testing and echocardiography is justified in children and adolescents; the patients may become symptomatic at any age. Indications for permanent pacing in pediatric patients have been difficult to determine due to the lack of data from controlled studies and multicenter trials. Pacing is not always easy in children. Epicardial pacing is advocated in small patients (< 5 years of age or 30 kg in weight). A pacing mode which restores the normal atrioventricular sequence is theoretically superior to single ventricular pacing even with rate responsive frequency. PMID- 11417314 TI - [Influence of placebo effect on planning and analysis of clinical trials]. AB - Placebos, seen as any pharmacological procedure without a specific effect, have been widely used in medical practice. In the fifties, when clinical trial development started, placebos assume a new dimension, as fundamental tools in clinical trial design. The aim of this review was to evaluate and describe methodological aspects related to placebo use in clinical trials. Despite the importance attributed to placebos, particularly to maintain blinding, it is not always easy to simulate an active procedure. On the other hand, the ethical aspects of placebo utilization in some clinical and experimental circumstances are becoming more and more complex. These two aspects are commented. It is emphasized that the use of a placebo, by itself, may influence the results of a clinical study. We analyze the factors that determine the placebo effect but also how placebos and the placebo effect affect final results. PMID- 11417315 TI - [Dobutamine stress echocardiography. Its contribution to the study of ischemic heart disease]. PMID- 11417316 TI - Quadricuspid aortic valve and atrial septal defect. PMID- 11417317 TI - Acute type A aortic dissection involving the left main coronary artery and the aortic valve--a case report. AB - The authors report a case study of a 60 year-old-male admitted to the Coronary Care Unit with severe chest pain, hypertension and ST depression on the anterior and lateral leads of the electrocardiogram. A diastolic murmur was heard in the aortic area. The chest X-ray showed an enlarged superior mediastinum. The transthoracic echocardiogram revealed an image that indicated an intimal flap above the aortic plane with severe aortic regurgitation. An aortography was performed and confirmed the diagnosis of acute type A aortic dissection with partial obstruction of the left main coronary artery, probably due to compression by the hematoma. The patient underwent emergency surgical repair with replacement of the aortic valve and ascending aorta. The patient survived without complications post surgery and was discharged ten days after onset of symptoms. Twenty months later, the patient was asymptomatic and the transesophageal echocardiogram showed a dissection of the descending thoracic aorta, mild aortic regurgitation and good left ventricular systolic function. PMID- 11417318 TI - [The Goldmann Memorial Lecture--historical and current aspects of stereopsis]. AB - According to Goldmann stereopsis is the highest performance of binocular recognition. A historical review shows that binocular stereopsis was explained only in 1838 by Wheatstone, whereas the monocular clues for stereopsis e.g. size, position and covering of the objects, light and shadow, perspective of the air, parallax and linear perspective were known long ago. The denomination depth perception is ambiguous: "depth" is the opposite of "height". Stereopsis means natural recognition of distance and objects in space, stereoscopy means recognition by the aide of an instrument. Natural and dichoptic stereopsis and the influences of vertical structures, of pupillary distance and of astigmatism are discussed. Examination of stereopsis is discussed, among others the two pencil test, the stereo-modification Bagolini-glasses, the combination of the plano-cylinders of W. R. Hess with random dots without glasses, the disk stereopsis etc. Goldmann's theory of binocular vision helps to understand different forms of strabismus, e.g. the statistical interplay of fixation and fusion for microtropia, the fixation mechanism for the congenital strabismus syndrome, the use of binocular stereopsis only for near explains intermittent divergence. PMID- 11417319 TI - [Glaucomatous optic neuropathy: a reperfusion injury]. AB - BACKGROUND: We introduce a pathogenetic concept which explains the present known clinical and experimental facts. MATERIALS AND METHODS: The known clinical risk factors for glaucomatous optic neuropathy as well as the outcomes of the experimental studies were systematically collected. RESULTS: IOP fluctuations are more damaging than a stable increased IOP, reduced circulation due to vascular dysregulation is more damaging than reduced circulation due to arteriosclerosis and the glaucomatous damage is clearly different from a pure ischemic lesion. CONCLUSIONS: Although IOP can damage mechanically, IOP fluctuation and blood pressure drops may lead to short term ischemia, followed by a reperfusion damage. A vascular dysregulation interferes with the autoregulation of ocular perfusion and renders the eye more sensitive to IOP increase or blood pressure decrease. PMID- 11417320 TI - [Diagnosis and treatment of intraocular tumors in the child]. AB - PURPOSE: To remind of the absolute necessity for early diagnosis in the presence of ocular signs in children giving rise to possible intraocular tumours. METHOD: Based on our own experience of intraocular tumours in children, together with findings from the literature, diagnostic criteria and methods of treatment are presented. RESULTS: Retinoblastoma is the predominant cause of intraocular tumours in children, representing over 80% of cases under the age of 15 years. Other diseases may give rise to the same initial signs, usually leukocoria, sometimes strabismus, more rarely other atypical signs. Elements taken into account for diagnosis include age, sex, laterality, heredity, size of the globe, clinical aspect of the tumours, presence of calcifications and vitreous seeding. Full fundus examination under general anaesthetic is usually necessary. Biological examination, ultrasonography, computerized tomography and MRI enable an accurate diagnosis to be made in the majority of doubtful cases. The management of retinoblastoma is adapted for each individual case from the wide range of treatments available. Enucleation, radioactive applicators (...), brachytherapy (...), cryo- and photocoagulation represent classical measures. Primary chemotherapy, combined with other treatments such as thermotherapy, has become the treatment of choice in those cases where external beam radiotherapy has been used up to now, or in some instances before enucleation. Enucleation is usually carried out for medullo-epitheliomas, but brachytherapy may offer an alternative. CONCLUSION: Any unexplained ocular sign in children should be considered as a possible retinoblastoma, making an accurate and certain diagnosis imperative. Early treatment may save not only the life but also the vision of patients carrying this highly malignant lesion. PMID- 11417321 TI - [Nonorganic disorders in ophthalmology: overview of diagnosis and therapy]. AB - BACKGROUND: Nonorganic diseases may be psychogenic (somatoform diseases) or due to malingering. Both occur frequently in the ophthalmologic practice. METHODS: This article reviews the different diagnostic and therapeutic procedures in nonorganic ocular diseases. RESULTS: After presenting the general symptoms, the origins of psychogenic diseases are discussed. Then, the specific tests used in ophthalmology to diagnose nonorganic diseases and the organic eye diseases most frequently confused with nonorganic diseases will be reported. At the end, we will discuss the treatment and the prognosis of psychogenic diseases. CONCLUSION: An early diagnosis of nonorganic diseases is important, since a delayed diagnosis may worsen their prognosis. PMID- 11417322 TI - [Non-steroidal anti-inflammatory drugs and ocular inflammation]. AB - BACKGROUND: Ocular inflammation is regulated by complicated mechanism that are responsible for the rupture of the blood-ocular barrier and the attraction of leukocytes towards the eye. This cellular trafficking is regulated by the liberation of inflammatory mediators and cytokines. MATERIALS AND METHODS: The main effects of prostaglandins that are described in the literature were reviewed. RESULTS: Despite the fact that the inhibitory action of aspirin and NSAIDs on prostaglandins secretion is known since 1971, it is quite recently that two isoenzymes of cyclooxygenase were discovered. The Cyclooxygenase-1 (COX-1) is constitutive and is responsible for the production of PGE2. This prostaglandin is related to the housekeeping functions such as gastric protection, platelet aggregation and maintenance of renal function. The expression of the cyclooxygenase 2 (COX-2) is induced by inflammation. The presence of COX-2 could be demonstrated in many ocular tissues (corneal epithelium and endothelium, pigmentary epithelium etc. ...). The use of NSAIDs in clinical practice allows a good control of ocular inflammation, of pain, of the maintenance of a good mydriasis during surgery and are impending the onset of postoperative cystoid macular edema. CONCLUSION: Prostaglandins are potent anti-inflammatory agents but their use is limited in the presence of severe dry eye syndrome or allergy towards NSAIDs. PMID- 11417323 TI - Lid retractors desinsertion in acquired ptosis and involutional lower lid entropion: surgical implications. AB - BACKGROUND: An association exists between upper and/or lower lid retractors' desinsertion and acquired ptosis and/or involutional lower lid entropion. METHOD: By highlighting the similarities that exist in the anatomy of the upper and lower lid retractors a possible pathophysiological mechanism leading to acquired ptosis and involutional lower lid entropion is suggested. As a logical consequence, in case of desinsertion, it is proposed to reattach the lid retractors to the tarsus. RESULTS: With this simple "physiological" surgical approach it is possible to treat with success a great majority of acquired ptosis and/or involutional lower lid entropion. CONCLUSION: Repairing upper and/or lower lid retractors' desinsertion can cure acquired ptosis and/or involutional lower lid entropion. PMID- 11417324 TI - [Treatment according to "evidence-based medicine" of aged-related exudative age related macular degeneration (laser photocoagulation and photodynamic therapy)]. AB - INTRODUCTION: Age related macular degeneration (AMD) is actually the leading cause of severe visual acuity loss in people over 65 in the Western World. Most of the visual loss is due to choroidal neovascularization (CNV). METHODS: Review of the literature. RESULTS: For more then 10 years laser photocoagulation was the only proven treatment for selected cases of AMD according to the evidence based medicine criteria. In 1999 photodynamic therapy was proven to be efficient for the treatment of predominantly classic subfoveal CNV. CONCLUSION: Laser photocoagulation and photodynamic therapy constitute therefore the only evidence based medicine treatment available for CNV in AMD. PMID- 11417325 TI - [GRIN (gradient-index) intraocular endoscopy--possibilities and limits--a review]. AB - BACKGROUND: Microendoscope techniques got a more important role in intraocular diagnostic and therapy. The used optical systems are fiberbundle-endoscopes and gradient-index (GRIN)-endoscopes. This study is concerned with the possibilities and limits of GRIN-endoscopes in pig eye models and in clinical use. PATIENTS AND METHODS: Several simulated intraocular techniques (phacoemulsification, implantation of intraocular lenses, vitrektomie, extraction of lost lens parts out of the corpus vitreous, extraction of metal foreign bodies) were done on pig eyes under GRIN-endoscopic control (GRIN-endoscope with diameter of 0.89 mm, Volpi, Switzerland). 3 clinical patients with cornea or lens opacities were examined. RESULTS: The image quality of simulated operations on pig eyes in clear intraocular conditions with a GRIN-endoscope was good. Also the clinical intraocular examinations could be done well, although the additional information were different. The in vivo examinations gave in one case no information (vitreous opacities very tight), in one case a relative information (mild vitreous opacities) and in one case an additional information (vitreous opacities rare). CONCLUSIONS: GRIN-endoscopes could complement the intraocular endoscopic spectrum in selected cases. At clear intraocular conditions good pictures were possible. GRIN-endoscopes could be used in cases where endoillumination is necessary. Then additional visual informations to the illumination are possible. The possibility to get overview images is limited with the actual endoscope probes. Ultrasound examination should be done in all cases with opacities of the anterior parts of the eye to get informations about the conditions inside the eye. Working inside the eye is possible with experience in this technique although a GRIN-endoscope has no stereopsis. Future developments should be done in stereopsis, several endoscope-tips and better handling (footpedal-controlling, autofocus). PMID- 11417326 TI - [Effect of squatting on sub-foveal blood flow defect in pseudophakic eyes operated by cerclage]. AB - ZIEL: To investigate the relationship between velocity (Velch), blood volume (Volch) and blood flow (Fch), and the mean ocular perfusion pressure (PPm) in the foveal region, and to determine how the regulatory capacity of the choroidal circulation is affected after an encircling buckle procedure. METHODS: We investigated both pseudophakic eyes of 6 patients (age range 56-79 years) in a masked study. Subjects presenting eye diseases (glaucoma, uveitis, diabetic retinopathy) as well as systemic diseases were excluded from the study. All subjects had in one eye a successful management of retinal detachment with an encircling buckling; the second eye was considered as control. Measurements of Velch, Volch and Fch were obtained by Laser Doppler Flowmetry (LDF) at baseline and during isometric exercise (squatting). RESULTS: In the operated eyes, Velch and Fch increased significantly (ANOVA, p < 0.05) during the PPm raise, which was not the case for Volch (ANOVA, p > 0.05). In the control eyes, all hemodynamic parameters remained unaffected by the PPm increase (ANOVA, p > 0.05). The response of Velch and Fch was significantly different (ANCOVA, p < 0.002) between operated and control eyes. CONCLUSION: These results suggest that encircling buckle does affect subfoveal choroidal blood flow regulation, which may explain a possible macular dysfunction in the operated eyes. PMID- 11417327 TI - [Albinism: diagnosis by visual evoked potentials]. AB - BACKGROUND: In albinism the majority of the nerve fibers of the optic nerve originating from an eye are innervating the contralateral hemisphere. As a result of the predominantly monocular innervation of the left and right hemispheres, the unilateral activation of the visual cortex (lateralization) can be detected with visual evoked potentials (VEP). PATIENTS AND METHODS: The VEPs were elicited with pattern onset stimulation. Five channels were recorded (2 channels over the left and right occipital lobe and 1 in the midline). 22 Patients with pendular nystagmus and 15 normal volunteers were examined. 5 of the normal volunteers were also examined with simulated pendular nystagmus (5-10 degrees, 1 Hz). The nystagmus was simulated by a simple galvanometer scanner based optical system. RESULTS: Unilateral activation of the hemispheres (lateralisation) was never detectable by normal volunteers. The lateralization of the VEP was detectable in all patients with oculocutaneous albinism with pattern onset stimulation. CONCLUSION: The pathological crossing of the nerve fibres in the optic chiasm is always detectable in oculocutaneous and ocular albinism with multichannel pattern onset VEP. The lateralization of the VEP is a more sensitive indicator of ocular albinism than other symptoms like macular hypoplasia, hypopigmentation, iris transillumination, nystagmus, reduced visual acuity et cetera. PMID- 11417328 TI - Decreased Heidelberg Retina Flowmeter (HRF) parameter "flow" at the papilla shortly after smoking a cigarette. AB - PURPOSE: To investigate the effect of smoking a cigarette on the Heidelberg Retina Flowmeter (HRF) parameter "Flow" at the papilla of healthy young volunteers. METHODS: Three HRF measurements were taken over the papilla (20 x 5, 256 x 64 pixels) in fourteen occasional smokers and in 14 non-smokers. Ten minutes later three similar HRF measurements were again made. In between the two series of measurements occasional smokers were asked to smoke a cigarette (Marlboro, nicotine content 9 mg) and non-smokers not. RESULTS: In occasional smokers, the values (arbitrary units) of the HRF parameter "Flow", calculated at the papilla (50 x 50 pixels), significantly (P = 0.01) decreased (11.2 +/- 3.5%) after smoking a cigarette. In contrast, in non-smokers, the values of the HRF parameter "Flow" did not decrease significantly (2.9 +/- 2.2%) when measurements were repeated 10 minutes later. CONCLUSIONS: The results of the present study suggest that smoking a cigarette can affect the HRF-parameter "Flow" at the papilla. The clinical implication of such an observation needs further investigations. PMID- 11417329 TI - [Value of indocyanine green angiography in detection of central serous chorioretinopathy]. AB - INTRODUCTION: Central serous chorioretinopathy (CSC) is a relatively common and self-limited disease affecting more commonly young adult males who are likely to have a type A personality. The aim of our study was to analyze indocyanine green (ICG) angiographic findings in CSC and correlate them with the biomicroscopical and Fluoresceine angiography (FA) findings in the affected and non affected eyes. PATIENTS AND METHODS: Our study included 19 consecutive patients, that were affected by CSC. The mean age of the patients was 44 years (range 35-50 years). In thirteen patients, in one eye typical acute lesions were observed, and in six patients presented chorioretinal inactive lesions. Angiographic ICG and FA findings of both eyes were compared. RESULTS: In 13 eyes (68%) presenting with the active form of CSC a classic focal leak with a smokestack phenomenon was seen in FA. The leaking point was identified in both FA and ICG, in 9 out of 13 eyes (70%). Surrounding the FA leaking point, an hyperfluorescent plaque in ICG was associated in 12 out of 13 (92%) eyes with active lesions. In 8 out of 13 (62%) controlateral eyes of this group, hyperfluorescent plaques in ICG were also observed. All eyes with inactive FA lesions (6 out of 6), expressed an hyperfluorescent plaque in ICG. DISCUSSION: ICG hyperfluorescence appears in the affected and the fellow eyes in areas where no clinical or FA signs of active disease were present. The persistence of abnormal ICG findings in all the cases of inactive disease suggest that CSC could be not a recurrent but a chronic disease. PMID- 11417330 TI - Vasorelaxing effect of the potassium (K+)-channel opener pinacidil in isolated porcine ciliary arteries. AB - PURPOSE: The aim of this study was to investigate the vasorelaxing properties of pinacidil (a potassium-channel-opener) in isolated porcine ciliary arteries. METHODS: Isometric contractions of isolated porcine ciliary arteries were measured with a myograph system. The vessels were first precontracted with the thromboxane A2 analog U 46619 (0.1 microM), and then exposed, in a cumulative manner, to increasing concentrations of pinacidil (1 nM-100 microM). Relaxations have been expressed in percent of the maximal contraction evoked by U 46619 (0.1 microM). RESULTS: Pinacidil showed a pronounced concentration-dependent relaxing effect in isolated porcine ciliary arteries. The difference between pinacidil and time-control experiments was significant (100 microM: 112 +/- 2% vs. 18 +/- 6%, P < 0.0001). The half-maximal concentration (pD50) value was 5.3 +/- 0.1 (-log M). CONCLUSIONS: The present study indicates that pinacidil has marked vasorelaxing properties in isolated porcine ciliary arteries. PMID- 11417331 TI - [Dependence of lipid peroxidation on pigmentation of the porcine iris]. AB - PURPOSE: Melanin has been shown to act as antioxidant in lipid peroxidation studies. We have now investigated lipid peroxidation in dependence on stromal pigmentation in isolated porcine irises. METHODS: The same number of lightly pigmented and heavily pigmented porcine irises (visual selection) were homogenized in buffer (50 mmol/l Na2HPO4, 50 mmol/l NaH2PO4 and 4 mmol/l sodium azide; 1:20 w/v). 500 microliters homogenate were incubated at 37 degrees C for 5, 10, 20 and 40 min in absence and presence of Fe2+ as inducer of lipid peroxidation. Lipid peroxidation was assayed by the thiobarbituric acid (TBA) test. Results are expressed as nmol of TBA reactive material produced (TBAR) per mg protein. Fe2+ concentration of the supernatant was determined spectrophotometrically with phenanthroline. RESULTS: 70 mumol/l, 180 mumol/l and 360 mumol/l Fe2+ induced lipid peroxidation. A plateau region was reached after 20 min. Lipid peroxidation differed in dependence on stromal pigmentation in porcine irises by a factor of 2.8. 180 mumol/l Fe2+ induced 1.373 +/- 0.138 nmol TBAR/mg protein in lightly pigmented irises compared to 0.491 +/- 0.125 nmol TBAR/mg protein in heavily pigmented irises after 10 min incubation (p < 0.0001, n = 4). On the other hand, the content of Fe2+ in the supernatant was the same within error. CONCLUSIONS: There was a stronger induction of lipid peroxidation in lightly pigmented porcine irises compared to heavily pigmented porcine irises. This effect may be related to the difference in stromal melanin content and its antioxidant activity. PMID- 11417332 TI - [Effect of chromatic flicker on circulation of the optic nerve]. AB - PURPOSE: To determine the response of human optic nerve head (ONH) blood flow (delta F) to heterochromatic equiluminant flicker modulation (eql-fl) and compare it to the response induced by pure luminance flicker (l-fl). METHODS: In 5 normal volunteers the ONH blood flow was measured by conventional laser Doppler flowmetry. Stimuli were generated by green and red light emitting diodes and delivered to the eye through a fundus camera illumination optic. Both green and red illuminances were square wave modulated in counter phase at different frequencies between 2 and 40 Hz. delta F was defined as the ratio between the ONH blood flow after 1 min stimulation and a baseline blood flow measured prior to the stimulation. RESULTS: In response to a 2 Hz eql-fl, ONH blood flow increases by 36% in average. delta F versus flicker frequency displayed the characteristics of a low-pass function with a cutoff frequency of 10 Hz for an eql-fl and a band pass function with broad maximum around 10 Hz for the I-fl. CONCLUSIONS: delta F in human ONH can be evoked by heterochromatic equiluminant flicker modulation. The blood flow frequency response to eql-fl and I-fl are similar to the neural activity dominated by the Parvo- and Magno-cellular activity, respectively. These findings offer a new approach to study the neurovascular coupling at the ONH in both physiological and diseased conditions involving predominantly or selectively the Magno- and Parvo-pathways. PMID- 11417333 TI - Brimonidine and inhibition of nitrite production in isolated porcine ciliary processes. AB - PURPOSE: To investigate whether the alpha 2-adrenoreceptor agonist brimonidine (a drug that decreases intraocular pressure in part by reducing aqueous humor production) can inhibit the production of nitrite induced by forskolin in isolated porcine ciliary processes. METHODS: Nitrite (a metabolite of nitric oxide) was measured (Griess reaction) before and two hours after exposure to the adenylcyclase activator forskolin (0.1 microM) in the presence or in the absence of the brimonidine (10 microM) and/or the alpha 2-adrenoreceptor antagonist yohimbine (10 microM). RESULTS: The increase of nitrite production evoked by forskolin (139 +/- 9%) was significantly (P < 0.01) inhibited by brimonidine (108 +/- 6%). The inhibitory effect of brimonidine on the increase of nitrite production induced by forskolin was significantly (P < 0.01) reversed by yohimbine (136 +/- 7%). CONCLUSIONS: The results of this pilot study suggest that brimonidine inhibits the production of nitrite evoked by forskolin through a mechanism that could involve alpha 2-adrenoreceptor activation in isolated porcine ciliary processes. PMID- 11417334 TI - Topical ocular instillation of nitric oxide synthase inhibitors and intraocular pressure in rabbits. AB - PURPOSE: It has been reported that intravenous injection of the nitric oxide synthase (NOS) inhibitor NG-nitro-L-arginine methyl esther (L-NAME) causes a rapid decrease of intraocular pressure (IOP) in rabbits. This study investigates the effect of topical ocular application of different NOS inhibitors on a raise of IOP induced by an acute water intake (rabbit water loading model). METHODS: Forty New Zealand (albino) male rabbits received randomly (within thirty minutes) in their right eye three 50 microliters installations of either 0.9% NaCl, 0.5% timolol maleate (beta-adrenoreceptor antagonist), 0.5% 7-nitroindazole (7-NI; NOS inhibitor), 0.5% L-NAME, or 0.5% 2-amino-5,6-dihydro-6-methyl-1,3,-thiazine (AMT; NOS inhibitor) before an oral water gavage (60 ml/kg). IOP was measured (TonoPen) before and after topical instillation (time 0), and then 15, 30, 60, 90, and 120 minutes after water intake. RESULTS: In right eyes, the area under the curve (AUC) of the IOP difference versus time (arbitrary units) was 527 +/- 284 for NaCl, 255 +/- 178 for timolol, 466 +/- 242 for 7-NI, 604 +/- 195 for L-NAME, and 394 +/- 202 for AMT. Values of AUC were only significantly lower (p < 0.05) in the timolol-treated group. In left eyes, no significant difference (p > 0.05) could be observed in values of AUC between groups. CONCLUSIONS: Under the present experimental conditions (including concentration and bioavailability of the drugs used), topical application of the NOS inhibitors 7-NI, L-NAME, and AMT does not prevent an IOP increase induced by water intake in rabbits. PMID- 11417335 TI - [Analysis of filtration according to implant type after deep sclerectomy in the rabbit]. AB - PURPOSE: The purpose of this study was to compare the aqueous humor outflow using three type of implants after deep sclerectomy. A numeric imaging system with ICG and fluorescein angiography was used. The evolution over time was recorded, while filtration and outflow facility were measured. MATERIAL AND METHODS: Numeric imaging system with ICG and fluorescein angiography were performed before and after deep sclerectomy at regular intervals after surgery. Both resorbable (collagen) and non-resorbable (hema) intrascleral devices have been tested in eight rabbits. The resorption time of the collagen implants has been monitored using UBM technique. The outflow facility was measured with the technique of anterior chamber canulation and constant pressure method. RESULTS: The resorbable collagen implants could be observed with UBM and angiography during the first six weeks post-operatively. The image of the non-resorbable Hema implant remained clearly visible over time. New drainage vessels were observed around the filtering bleb. The filtration significantly increased as the outflow facility improved after surgery, while the intraocular pressure remained stable over time. CONCLUSION: This conjunction of UBM, angiography and outflow measurement allowed us to study and compare three types of implants used as space maintainer after deep sclerectomy, and analyse their effect on the aqueous filtration and outflow facility. PMID- 11417336 TI - [Necessary visual information for restoring reading with a retinal implant in a blind patients with massive retinal degeneration of photoreceptors]. AB - DEFINITION OF THE PROBLEM: Our goal is to determine the minimum of information necessary for elementary reading, using a retinal implant. This concerns particularly the fragmentation (pixellisation) of the presented image and its position in the visual field. Fragmentation corresponds to the number of electrodes available, the position of the image in the visual field is equivalent to the site of the implant on the retina. MATERIAL AND METHODS: 10 degrees x 10 degrees windows, containing isolated words or letters, were presented to six healthy subjects on a computer screen. A coupling between the computer and an eye tracker stabilizes these images in an area of the visual field. This coupling constantly corrects the position of the image on the screen according to the direction of gaze. RESULTS: 1) A rapid decrease of the performance is observed at a certain threshold of pixellisation, dependent on the eccentricity of presentation of the images. 2) In central vision, about 400 pixels are sufficient to recognize 80% of the four-letters words. At 10 degrees of eccentricity, about 1225 pixels are needed. 3) An acceptable comprehension of a text (identification of four words out of five), is impossible at eccentricities higher than 10 degrees 4) About 50 pixels are sufficient for a satisfactory recognition of isolated letters, independently of their eccentricity. CONCLUSION: These data validate the method of investigation and provide valuable indications regarding minimal visual requirements in prosthetic vision. PMID- 11417337 TI - Modulation of Na/K-ATPase activity by isoproterenol and propranolol in human non pigmented ciliary epithelial cells. AB - PURPOSE: The present study investigates whether beta-adrenoreceptor agents such as isoproterenol and propranolol can regulate Na(+)-K(+)-ATPase activity in cultured human non-pigmented ciliary epithelial cells. METHODS: Human non pigmented ciliary epithelial cells (ODM2) were grown to confluence. The active ion transport mediated by the Na(+)-K(+)-ATPase was evaluated by measuring ouabain-sensitive rubidium (Rb+) uptake. In a first set of experiments, cells were exposed to the beta-adrenoreceptor agonist isoproterenol (0.01-10 microM). In a second set of experiments, cells were exposed to isoproterenol (1 microM) in the presence of different concentrations of the beta-adrenoreceptor antagonist propranolol (0.01, 0.1, 1 microM). RESULTS: In a concentration-dependent manner, isoproterenol induced an increase in Na(+)-K(+)-ATPase activity. The maximal Na(+)-K(+)-ATPase activity was observed at a concentration of 1 microM of isoproterenol (283 +/- 58%, P < 0.001). The increase in Na(+)-K(+)-ATPase activity evoked by isoproterenol (1 microM) was inhibited. In a concentration dependent manner, by propranolol (maximum: 659 +/- 39 vs. 141 +/- 42 pM/mg protein/min, P < 0.01). CONCLUSION: The beta-adrenoreceptor agents isoproterenol and propranolol are apparently able to modulate Na(+)-K(+)-ATPase activity in cultured human non-pigmented ciliary epithelial cells. PMID- 11417338 TI - L-NAME- and U 46619-induced contractions in isolated porcine ciliary arteries versus vortex veins. AB - PURPOSE: To investigate contractions evoked by the thromboxane A2 analog U 46619 and by the inhibitor of nitric oxide formation NG-nitro-L-arginine methyl ester (L-NAME) in isolated porcine ciliary arteries and vortex veins. MATERIAL AND METHODS: In a myograph system (for isometric forces measurement), vessels were exposed (at different levels of wall tension) to 100 mM potassium chloride (KCl). At their optimal tension, vessels were exposed (in a cumulative manner) to increasing concentrations of U 46619 (0.1 nM-1 microM) in the absence or in the presence of L-NAME (0.1 mM). Contractions were expressed in mN or in percent of a 100 mM KCl-induced contraction. RESULTS: Optimal tension was higher in arteries (7 mN) than in veins (3 mN). Maximal contractions induced by KCl were stronger in arteries (24.4 +/- 3.6 mN; n = 8) than in veins (1.8 +/- 0.2 mN; n = 8). In contrast, maximal contractions evoked by U 46619 were proportionally higher (p < 0.001) in veins (178.3 +/- 8.9%; n = 8) than in arteries (108.4 +/- 2.6%; n = 5) and were not significantly affected by L-NAME. Sensitivity to U 46619 was not significantly different between arteries (pD50 = 7.7 +/- 0.1) and veins (pD50 = 7.9 +/- 0.1). In quiescent vessels, L-NAME evoked contractions that were higher (p < 0.001) in veins (43 +/- 7.9%; n = 13) than in arteries (7.5 +/- 1.7%; n = 10). CONCLUSIONS: When compared with KCl-induced contractions, contractions evoked by U 46619 or L-NAME are proportionally higher in porcine vortex veins than in ciliary arteries. PMID- 11417339 TI - Cyclic AMP and anionic currents in porcine ciliary epithelium. AB - INTRODUCTION: To investigate whether in the ciliary epithelium of isolated porcine ciliary body cyclic 3',5' adenosine monophosphate (cAMP) activates transmembrane anionic currents. METHODS: Changes in membrane potential induced either by the adenylcyclase activator forskolin (10 microM; n = 4) or the stable membrane permeable cAMP analog 8-bromo-adenosine 3',5'-cyclic monophosphothioate (8-br-cAMP; 30 microM; n = 4) were measured with intracellular microelectrodes. The effect of the drugs were assessed in the absence or in the presence of the non-selective anionic channel/transporter inhibitor diisothiocyanatostilbene-2,2' disulfonic acid (DIDS; 1 mM; n = 4). RESULTS: Significant (p < 0.001) membrane potential depolarization were induced by both forskolin (11.8 +/- 0.3 mV) or 8-br cAMP (9.3 +/- 0.4 mV). In the presence of DIDS, a significant (p < 0.001) inhibition of the depolarization evoked by forskolin (0.9 +/- 1.1 mV) and 8-bromo cAMP (0.7 +/- 0.2 mV) was observed. CONCLUSIONS: In the ciliary epithelium of isolated porcine ciliary body cAMP induces membrane potential depolarization through a process that could involve anionic channels. PMID- 11417340 TI - Latanoprost is a vasoconstrictor in isolated porcine ciliary arteries. AB - PURPOSE: The purpose of this study was to assess the vasoconstrictor effect of the prostaglandin F2 alpha analog latanoprost in isolated porcine ciliary arteries. MATERIAL AND METHODS: With a myograph system to measure isometric forces, porcine ciliary arteries were exposed, in a cumulative manner, to increasing concentrations of latanoprost (0.1 nM-0.1 mM). Experiments were conducted in quiescent vessels as well as in vessels precontracted with potassium chloride (KCl; 40 mM) or endothelin-1 (10 nM). Contractions were expressed in percent of 100 mM KCl-induced contraction. RESULTS: In a concentration dependent manner latanoprost induced significant (p < 0.001) contractions in quiescent vessels (maximum: 68 +/- 3%, n = 5; PD50: 5.7 +/- 0.1). In KCl or in endothelin-1 precontracted vessels latanoprost did not evoke any relaxation but in contrast significant (p < 0.001) contractions (maximum: 33 +/- 4%, n = 5 and 26 +/- 6%, n = 8, respectively). CONCLUSIONS: Latanoprost has a vasoconstrictor effect in isolated porcine ciliary arteries. No relaxation could be observed. The clinical relevance of this finding requires further evaluation. PMID- 11417341 TI - [Technical construction, calibration and results with a new intraocular pressure sensor with telemetric transmission]. AB - BACKGROUND: Presently intraocular pressure is measured indirectly by applanation or impression of the cornea. Only isolated values are available with this method. We present a new implantable system for direct and continuos measurement of the intraocular pressure. METHODS: An implantable system consisting of a miniaturized sensor and a telemetric unit was integrated in an intraocular lens. The eye pressure is determined by the sensor, modulated and transduced by the telemetric system. By an extracorporal receiver the signal is demodulated. The electric supply of the intraocular system is achieved by external electromagnetic induction. RESULTS: The telemetric transmission of the intraocular pressure can be achieved with an accuracy of +/- 1 mm Hg and a frequency of registration of 3 values per second. CONCLUSION: Clinical application necessitates further animal trials in vivo. PMID- 11417342 TI - Pupillary examination with infrared consumer videocamera. AB - BACKGROUND: Pupillary observation in the dark is always a problem in a general ophthalmological practice or an outpatient clinic without specialized equipment. We present two methods for observation of the pupils in darkness: 1) illumination of the pupils with the skiascope as a routine examination and 2) infrared observation of the pupils with a consumer digital video camera. METHODS: (1) Pupillary reactions are observed with the skiascope/retinoscope, the observation beam of the device focused to infinity and documented with a video camera. (2) Infrared observation of the pupils was performed with a digital consumer video camera, allowing observation of the pupillary reaction in darkness. After recording, video sequences of interest were transferred to a personal computer and the still images of interest extracted. RESULTS: In everyday clinical routine, observation of the pupillary reaction with the skiascope/retinoscope proves as a reliable tool with a large bandwidth of illumination and a high contrast between pupil and surrounding area. The infrared video camera allows an excellent visualization of the pupillary reflex in darkness. The transfer of the video sequences to a personal computer proved to be simple and single images can easily be chosen. CONCLUSION: Observation of the pupillary reflex with the skiascope proved a useful tool that is available in practically every ophthalmological office. Use of the infrared digital consumer video camera (available at low prices) is a highly sophisticated tool for observation and documentation of pupillary reflex in darkness. PMID- 11417343 TI - Lacrimal drainage system endoscopic examination and surgery through the lacrimal punctum. AB - BACKGROUND: To describe an approach to restore the physiological tear passages of an obstructed lacrimal drainage system with instruments introduced through one of the lacrimal punctum. METHOD: In a first step a miniature endoscope (O: 0.9, 1.1, 1.3 mm) is introduced in one of the lacrimal punctum to visualize the level, the extent, and the nature of an obstruction along the lacrimal drainage system. In a second step, the miniature endoscope is replaced by a lacrimal trephine (O: 0.9, 1.1, 1.3 mm) that holds an optic fiber in its lumen, and the obstruction is removed under endoscopic visual control. The entire procedure is performed under local anesthesia. RESULTS: With this approach it is possible to visualize and remove partial or total fibrous obstructions along the lacrimal drainage system (lacrimal canaliculus, nasolacrimal duct). Lacrimal sac lithiasis can also be fragmented and eliminate in the nose through the nasolacrimal duct. During or after the procedure only minor, and no major, complications can sometime occur, such as hematoma or edema (after lacrimal irrigation) of the surrounding soft tissues. CONCLUSION: The approach described here is straight forward, quick, and can be performed under visual endoscopic control and in local anesthesia. With this approach partial stenosis can easily be diagnosed and cured. This procedure might potentially change our current concepts regarding surgical indications for obstructive epiphora and dacryocystitis. However, the long-term results of this type of surgery need to be challenged. PMID- 11417344 TI - [Formation of a descemetic bleb after deep sclerectomy and visco-canalostomy: apropos of 2 cases]. AB - BACKGROUND: Deep sclerectomy with visco-canalostomy is a new non perforating method for the surgical treatment of glaucoma. PATIENTS AND METHODS: We report 2 patients who developed a descemetic bleb after this surgical procedure. RESULTS: The cornea remained clear for more than ten months, as long as the bleb was not ruptured. In case of rupture, one should inject air into the anterior chamber to fix the descemetic membrane, otherwise the cornea will become quickly cloudy. CONCLUSIONS: Descemetic bleb formation after visco-canalostomy is a new complication which has to be recognized and treated in order to avoid corneal edema. PMID- 11417345 TI - [Masquerade of sebaceous gland carcinoma as a rapidly recurring "chalazion", a case report]. AB - PURPOSE: Demonstration of the importance of surgical excision and histological examination in presence of an apparently harmless tumoral alteration of the eyelids. CASE PRESENTATION: We report the case of a 65 year old patient suffering from systemic lupus erythematosus who noted a tumoral lesion on his left lower eyelid. Suspecting a chalazion, his dermatologist simply performed a cauterization. Six weeks later, a recurrence of the tumor appeared at the same location, and again, cauterization was done. A few weeks later, the patient consulted our clinic with a polycyclic tumor of 5 mm in diameter, involving the lid margin of the temporal part of the lower left eyelid. The patient had only moderate signs of blepharitis. There were no palpable preauricular and cervical lymph nodes. Suspecting a malignant tumor, the entire tumoral lesion was removed surgically. HISTOPATHOLOGY: The histopathologic examination showed a highly differentiated sebaceous gland carcinoma, most probably originating from a meibomian gland. The margins of the excision were found to be tumor-free. DISCUSSION: Sebaceous cell carcinoma is a rare entity. Depending on its histological differentiation it can be highly malignant. Infiltrative, and can metastasize. The mortality may reach 30% if low differentiation is present. As illustrated in the present case, the lesion may masquerade a chalazion. Therefore, in case of atypical lesion of the eyelid region complete surgical removal followed by a histopathological examination should be performed. PMID- 11417346 TI - Immunosuppressive therapy in serpiginous choroiditis--case report and brief review of the literature. AB - BACKGROUND: At present there is no consensus regarding the efficacy of the immunosuppressive therapy in serpiginous choroiditis. PATIENT AND METHODS: We describe the case of a patient with a macular serpiginous choroiditis. Despite corticosteroid therapy the choroiditis was progressive and threatened the central vision. Therefore a triple immunosuppressive therapy with prednisone 1 mg/kg body weight/day (mg/kg/d), azathioprine 1.5 mg/kg/d, and low-dose cyclosporine A 4.5 mg/kg/d was introduced. RESULTS: Under this triple immunosuppressive therapy a prompt stabilisation of the chorioretinal process could be observed in fluorescein angiography. After five months of therapy the examination of the visual field shows an impressive recovery. No significant side effects have been noted. CONCLUSIONS: In the literature, until now only six patients with serpiginous choroiditis treated with a triple agent immunosuppressive therapy have been reported. In all cases this regimen seemed to have a positive influence on the disease. Our present observation corroborates the beneficial effect of a triple agent immunosuppressive therapy on the course of serpiginous choroiditis. However, the beneficial effect of such a therapeutic approach needs to be confirmed by further observations. PMID- 11417347 TI - [Endogenous fungal endophthalmitis: results of antifungal treatment with and without vitrectomy]. AB - BACKGROUND: Fungal endophthalmitis represents a significant cause of ocular morbidity, affecting in the majority of cases patients in poor general conditions. The eye is typically involved by hematogen dissemination, and the germ induces a chorioretinitis associated with an important panuveitis. PATIENTS AND METHODS: Four patients were examined. They complained of a progressive reduction of vision, associated with photophobia. Ophthalmologic examination disclosed an important panuveitis. Investigations showed a fungal chorio retinitis in all cases. Risk factors were intra-venous toxicomania, longterm parenteral nutrition and traumatism of the sinuses. RESULTS: Vitreous cultures were positive for Candida albicans (3 patients) and for Aspergillus fumigatus (1 patient). Blood cultures were negative in the four cases. Three patients were treated with anti-fungal medication (fluconazole, itraconazole) associated with a vitrectomy by pars plana. One patient was treated by anti-fungal therapy only. Clinical evolution was satisfactory in all cases. Final vision was 10/10 in three cases and 5/10 in one. One patient developed a retinal detachment and an epiretinal membrane. Follow-up was 7 months (2-16 months). CONCLUSION: The evolution of these four cases suggests that a rapid anti-fungal therapy associated with or without a vitrectomy represent a favourable therapeutic option when a fungal infection is suspected. PMID- 11417348 TI - [A free ride for free expert witnesses]. PMID- 11417349 TI - [Open angle glaucoma: risk factors and progression]. PMID- 11417350 TI - [Effectiveness of intraocular polyacrylate lenses]. PMID- 11417351 TI - [Retinitis pigmentosa: a report of experiences]. PMID- 11417352 TI - [Industry-promoted research and the significance principle]. PMID- 11417353 TI - [Efficient personnel politics for the physician's practice]. PMID- 11417354 TI - [The physician's practice and employment income. Questions about the medical employment center]. PMID- 11417355 TI - [Psychotherapists to prison!]. PMID- 11417356 TI - [America a shining example? Developments in medicine between brain, mind and money]. PMID- 11417357 TI - [Anxiety and depression in the general population: normal values in the Hospital Anxiety and Depression Scale]. AB - For the Hospital Anxiety and Depression Scale (HADS) psychometric properties were tested and standardised values were calculated on the basis of a representative sample of the German adult population with 2037 persons. The main result was the evidence of age and gender differences for anxiety and depression. Females were more anxious than males. For both dimensions of the HADS a nearly linear age dependency was found which was more pronounced for depression (r = 0.36) than for anxiety (r = 0.14). Standardised values are given for different age and gender groups, and the results of regression analyses are presented. The psychometric properties were satisfying or good, the two-dimensional factorial structure could be replicated. By means of the standardised values and regression coefficients it is now possible to compare patient groups of different age and gender distributions with the general population. PMID- 11417358 TI - [Development of the Borderline Symptom List]. AB - The categorical diagnosis of borderline personality disorder (BPD) is currently discussed as sufficiently reliable and valid. However, specific standardized instruments to measure the degree of the symptomatology are currently not available. Based on the criteria of DSM-IV, the Diagnostic Interview for Borderlines (DIB-R) and the opinion of clinical experts and borderline-patients, a self-rating scale was developed to quantify the intrapsychic strain of patients with BPD. 308 female borderline-patients scored the 99 items. Factor analyses revealed seven factors: self-image, affect regulation, autoaggression, dysthymia, social isolation, intrusions and hostility. Analyses of reliability and validity revealed promising results. PMID- 11417359 TI - [A combination of cognitive and psychodynamic components in the psychotherapy of somatoform disorders]. AB - Psychological research has identified difficulties in the awareness, verbalization and recognition of emotions in patients with somatoform disorders. Problems in the recognition of emotions are usually labeled as alexithymia. Patients with alexithymic characteristics need a special psychotherapeutic approach, which may be described as follows: The first stage of treatment aims to overcome the alexithymic barrier along with the development of self-monitoring skills for emotional states. A favorable setting for this work is group psychotherapy. In this stage of treatment cognitive targets and interventions dominate. These include: 1. the link between emotions and physiological response; teaching the fundamental laws of emotional life and the importance of emotional hygiene. 2. Improving awareness of difficulties in the recognition and expression of emotions. 3. Changing the patients negative attitude towards emotions. The second stage of the group work is mainly oriented towards the cognitive analysis of dysfunctional basic beliefs. In this stage psychodynamic issues become more important which include resistance and transference reactions, feedback of group members, analysis of social and familial contexts and the working through of traumatic childhood experiences. PMID- 11417360 TI - [Psychotherapeutic resources in the General Hospital of Vienna. A representative empirical study of psychotherapeutic practices]. AB - Since 1993 the Austrian Federal-Hospital-Law demands the guarantee of adequate psychotherapeutic and clinical psychological care in Austrian hospitals. The target of the present survey was to explore the extent of realisation of this demand concerning the psychotherapeutic care in the General Hospital of Vienna (AKH). 125 of the 128 employees with psychotherapeutic qualifications were interviewed. The considerable number of psychotherapeutically qualified employees cannot obscure the fact that inadequate structural preconditions impede a satisfying care. These are: most of the interviewed persons have been employed because of other vocational qualifications (medical doctors, psychologists etc.). Therefore they can only use a very limited part of their worktime on psychotherapeutic interventions. Deficits of spatial and temporal resources could be detected as well as unclear vocational characteristics and qualification requirements. These deficits point out the necessity of developing professionally founded organisation structures, which enable well aimed networking and an optimal usage of resources. PMID- 11417361 TI - [Three therapeutic processes of inpatients with eating disorders compared with the Core Conflict Relationship Method (CCRT)]. AB - In this article the results of a research with the CCRT-method on the psychotherapies of three in-patients suffering from eating disorders are presented. The CCRT describes recurrent internal and interpersonal relationship patterns in narratives. Each therapy session was recorded on tape and transcribed. The CCRT components were identified from the verbatim transcripts. All three patients clearly showed a negative self-perception and despite a more graded attitude regarding other people they felt rejected by the community throughout the entire therapy. The CCRT of each patient was different: patient 1. had conflicts between dependence and independence with increasing autonomy; patient 2. had great symbiotic desires, which at the beginning of the therapy were warded off with a performance ideal; patient 3. showed self-assertion and an increasing level of openness against the community, despite a high level of fear and self-isolation at the onset of therapy. Despite methodological deficiencies the CCRT method proved to be sensitive enough to show similarities and differences among the individual courses of treatment. PMID- 11417362 TI - [Value of ultrasound diagnosis in pediatrics]. AB - Due to its easy applicability, lack of invasiveness, and high yield of information, ultrasound performed by the experienced examiner has become an indispensable diagnostic tool in many areas of clinical pediatrics. This is reflected by the large numbers of ultrasound studies both in pediatric private practice and in children's hospitals. Therefore, quality standards and quality control are of paramount importance. For many clinical problems, ultrasound has led to new diagnostic and even therapeutic concepts. In the German-speaking countries, ultrasound has officially been established as a screening method for the hip, but in clinical practice also for other organ systems. Due to technological developments of the last few years, the diagnostic yield has significantly been improved, and new areas of application have been established. PMID- 11417363 TI - [Value of pneumococcal vaccination in infants and young children]. AB - Streptococcus pneumoniae is a common cause of meningitis, sepsis, pneumonia, acute otitis media, and sinusitis in children. Children younger than 24 months have the highest rates of invasive pneumococcal infections (Germany 1997-1999: 19.5/100,000/year). Pneumococcal infections cause in Germany 220-250 cases of meningitis, about 50,000 of pneumonia (children younger than 5 years) and more than 1 million cases of otitis media (children) annually. The case-fatality rate for invasive pneumococcal diseases is high (1997-1999 5.5%, meningitis 8.3%). 20 30% of survivors from meningitis suffer from CNS-related sequelea. In children up to 2 years vaccination with the heptavalent pneumococcal conjugate vaccine can reduce invasive pneumococcal diseases by about 80% and otitis media and recurrent otitis media by 6% and 9-16%, respectively. Due to the increased risk of pneumococcal infections in the first two years of live all children of this age group should be vaccinated. The high rate of resistance of pneumococci against macrolides in Germany, the high rate of non-licensed antibiotics in infants and the inefficacy of the 23-valent vaccine in children younger than 2 years makes the new vaccine to a necessary alternative. PMID- 11417364 TI - [Polysomnography in infancy--necessary or expensive luxury? Therapeutic and interventional consequences]. AB - An increasing amount of polysomnographic studies are performed in early infancy. Complete pediatric sleep laboratory systems are commercially available and provide fast and exhaustive results if operated by trained personnel. Experience in evaluation is necessary, because a broad range of possible diagnostic findings exists, and artifacts are common. Based on longitudinal studies in 31 healthy infants, we present reference values for several polysomnographic parameters. Aberrant results must be interpreted in the context of patient's history, clinical state, and additional laboratory findings. In presence of a clearly defmed indication for polysomnographic testing, clear therapeutic and interventional strategies can be derived. Possible consequences include the stimulation of the respiratory drive by drugs with subsequent polysomnographic control of therapeutic effect, a transfusion, the prescription of a home monitor, and the recommendation of further diagnostics or of control polysomnographies. In conclusion, infant polysomnography is an important diagnostic tool in pediatrics. PMID- 11417365 TI - [Ischemia of the brain stem caused by compression of the vertebral arteries by head rotation--an etiology for SIDS?]. AB - Hypoperfusion of the brainstem during head rotation may be a risk factor for the development of SIDS. On this background we established a Doppler sonographic screening programme of the basilar cerebral arteries to evaluate the dependency of blood flow on head and body position. PATIENTS AND METHOD: We investigated 3840 newborns (1872 girls and 1968 boys) with a birth weight of 3399 +/- 497 g and a gestational age of 39.2 +/- 1.4 weeks. The investigations were performed in the neonatal period with an average age of 4.7 +/- 3 days. In all infants blood flow was measured in the basilar artery (BA) in supine position with the head in the midline. From the flow profile peak systolic flow velocity Vs and time average flow velocity TAV were measured. Additionally flow measurements were performed in supine and prone position with rotation of the head to the right and left side. A decrease of blood flow velocities below 50% of the value in neutral position was considered to be abnormal. Retrograde or biophasic flow profiles during rotation were considered to be pathologic. In infants with abnormal or pathologic flow during rotation of the head flow measurements in the vertebral arteries (VA) were additionally performed. Blood flow velocities in the VA were measured in supine and prone position with the head in the midline position and after rotation to the right and to the left. In neutral position unilateral vertebral hypoplasia, aplasia and normal VA were differentiated. The judgement after rotation was performed such as in the BA. RESULTS: In 3807 infants (99.14%) blood flow velocities during head rotation did not decrease below 50% of the value measured in neutral position. In 33 infants (0.86%) a decrease of blood flow velocities below 50% could be found during rotation. In 7 infants (0.18%) a pathologic flow could be found during head rotation. 27 of the 33 infants with abnormal and pathologic blood flow in the BA during rotation showed anatomic abnormalities of the VA. 20 of these infants (61%) had unilateral vertebral hypoplasia (11 right, 9 left side), 7 (21%) had unilateral vertebral aplasia (4 right, 3 left side). 32 of the 33 infants with abnormal flow in the BA showed a decrease of blood flow in the contralateral VA during head rotation. 9 infants had an abnormal, 19 a pathologic flow within the contralateral VA. In 4 infants the corresponding VA could not be measured during head rotation. The decrease of blood flow velocities in the BA during head rotation was caused by compression of the contralateral VA at the craniocervical junction. CONCLUSION: Blood flow within the basilar artery of healthy infants is independent of body position and rotation of the head. A decrease of the flow velocities below 50% during rotation has to be considered as an abnormality. The incidence of pathologic blood flow during head rotation with 1.8@1000 approximates the incidence of SIDS. Hypoperfusion of the brainstem during head rotation may be a risk factor of SIDS. PMID- 11417366 TI - [Cockayne syndrome with marked cerebral symptoms]. AB - The course of Cockayne syndrome is reported in two sisters over a period of 14 years. Both girls developed characteristic clinical signs early. Reaching the second decade progeria and psychomotor deficits progressed quickly with a marked mental decline brought about by the cerebral destruction which is demonstrated by successive CT und MRI scan. The effects of defective DNA repair mechanisms on progeria and mental deterioration are discussed and differential diagnoses are shown. PMID- 11417367 TI - [Nasal glioma as a rare cause of obstructed nasal breathing in a newborn infant]. AB - Congenital midline nasal masses are rare entities occurring once in 20-40,000 births. They are often misdiagnosed or even missed. These tumors are most commonly present in newborn infants and children, but rarely they can be discovered in adults as well. The differential diagnostic possibilities are dermoids, gliomas, encephaloceles and epidermoid cysts. Although they are benign, they have the potential for disfigurement, destruction and causing a meningitis, therefore they require prompt diagnosis and management. Because of a possible connection to intracranial structures a complete radiologic evaluation is essential. The treatment for these nasal masses is surgical resection, endoscopically controlled procedures are preferred. We report the case of a newborn with respiratory distress shortly after birth. The infant was discharged from the hospital, because the unexplained symptoms were not progressive. After a few days the child was send to an otolaryngologist, who found a suspicious mass near the top of the nose and performed a biopsy. At 20 days of age the infant was brought to our children's hospital for additional diagnostic procedures and therapy. The MRI showed a soft tissue mass (1.2 x 0.7 x 1.1 cm) in the nasal cavity. There was no clear-cut evidence of an intracranial extension, for further evaluation a radionuclide scan was performed. This confirmed the integrity of the base of the skull. 6 weeks after the birth a smooth tumor was successfully removed by an endoscopically controlled operation. Histopathologic studies confirmed neuroglial tissue. Because of ist rare incidence, many physicians are not familiar with the diagnosis and management of these tumors. Respiratory abnormalities in newborn should always lead to a careful physical examination and complete radiologic evaluation, i.e. MRI, CT and radionuclide scan. PMID- 11417368 TI - [Clinical aspects and genetics of Prader-Willi syndrome]. AB - Prader-Willi syndrome (PWS) is considered to be a rare neurogenetic disorder, nevertheless it represents the most common syndromatic obesity. Main features are severe hypotonia in the newborn period with feeding difficulty and failure to thrive in the first few months, and improvement in later infancy. Between 1 and 6 years the development of a marked truncal obesity is observed, sometimes later on reaching a life-threatening degree. Apart from retarded motor development speech ability is also hampered due to dysfunction of oropharyngeal muscles. Moreover, intellectual impairment is observed that leads to mostly moderate learning difficulty due to deficits in short term memory and abstract thinking. The genetic background of PWS is loss of function of a paternally inherited gene cluster on chromosome 15q11.2, therefore representing a paradigm of an epigenetic phenomenon with silencing and activating of genes depending on their parental origin. Together with the Angelman-Syndrome (AS), genetically located in the same region but clinically different PWS was identified as one of the first human disorders to be caused by the mechanism of genomic imprinting. There are some different genotypes in PWS leading to a quite similar phenotype with small differences e.g. in the pigment expression. An early diagnosis is important because the neuromuscular dysfunction improves with appropriate physiotherapy. Even more, dietary programs with periodical calorie restricted meals can counteract development of morbid obesity with subsequent complications of cardiovascular disorders and diabetes. Clinical diagnosis remains difficult especially in the newborn period and is considered mostly because of the marked hypotonia. Today, the availability of molecular testing of loss of function in the paternal inherited PWS candidate gene makes a definitive diagnosis possible as a prerequisite of symptomatic therapy. One of the most recommended therapeutic interventions is the application of recombinant human growth hormone (rhGH) which has been shown to be useful for improvement of length, physical ability and favourable influence on respiratory problems by a lot of clinical studies. SUMMARY: Early diagnosis by means of molecular methods is helpful for comprehensive genetic counseling. It may avoid unnecessary investigations like computed tomography of the brain and muscle biopsy and it enables parents and professionals to start a purposeful therapy. Although it remains a symptomatic tool, GH therapy appears to be useful for most of the patients. Considering the multimorbidity in PWS a multidisciplinary approach seems appropriate for this still mystical condition. PMID- 11417369 TI - [Minimal invasive pediatric surgery: development and progress by innovative technology]. AB - BACKGROUND: Minimally invasive pediatric surgery (MIPS) has a solid tradition in which technique and technology have made key contributions to an already broad range of indications. This surgical method still has a deficiency with regard to tissue management of large-area defects, however. Sealing techniques can further expand the range of application. METHOD: Technological evaluation provided outstanding data of fleece-bound collagen- and fibrinogen-based sealing systems (TachoComb) on biodegradability, adhesive strength and practicability. A relevant instrument was developed for MIS application and was introduced as the ATCS (AMISA-TachoComb-System). PATIENTS: From 1993-2000, ATCS sealing was carried out in the scope of thoracoscopy (105 procedures) and laparoscopy (53 procedures) and specifically for recurring pneumothorax, traumatic chylothorax and splenic trauma. RESULTS: Pneumothorax: 59 ACTS procedures in 49 patients (mean age: 11.4 yrs) with 6 reoperations (10.2%) and one recurrence (1.7%). The drainage dwell time was reduced (p < 0.05) using a conventional comparison (31.9 hours vs. 17 days) and further relevant parameters were also reduced. Chylothorax: 3 ATCS procedures in 3 patients (mean age: 6.3 years) with reduction in the drainage dwell time (p < 0.05) based on a conventional comparison (35 hours vs. 18 days). Splenic trauma: 17 ATCS procedures in 16 patients (mean age: 8.9 years) with one re-operation (5.9%) for associated liver trauma, organ conservation in each case and no significant drainage volumes. CONCLUSION: The ATCS is an innovative instrument for MIPS and ca be employed for efficient and socio-economic (e.g. DRGs) closure of large-area defects. PMID- 11417370 TI - Vesicles on the extremities. Patients who spend time outside may be especially prone to these lesions in the summer. PMID- 11417371 TI - Type 2 diabetes. How new insights, new drugs are changing clinical practice. AB - In 1997, the American Diabetes Association recommended a normal fasting blood glucose of < 126 mg/dL as the criteria for diagnosis of type 2 diabetes. Since then, new data have suggested that post-prandial glucose may have a stronger correlation with cardiovascular disease than fasting blood glucose. Two trials, the DCCT and UKPDS, provided evidence of the relationship between hyperglycemia and long-term diabetic complications. Preventing short-term complications, such as cognitive decline, is a more immediate goal and less well-studied. Type 2 diabetes is understood to result most often from insulin resistance and insulin deficiency. New classes of drugs offer expanded therapeutic options for managing this dual metabolic defect. These drugs have invalidated the former therapeutic paradigm of diet, sulfonylureas, and then insulin therapy. PMID- 11417372 TI - Multiple 'chief' complaints. Negotiation is the key to prioritizing office visits and avoiding last-minute lists. PMID- 11417373 TI - Nocturnal leg cramps. Clinically mysterious and painful--but manageable. AB - Nocturnal leg cramps are common occurrences among older, generally healthy adults. Although there are many known causes--endocrinologic, neurologic, and vascular disorders, treatment with certain drugs, and occupational factors--a significant portion of cases are idiopathic. History, physical exam, and laboratory testing can provide clues for distinguishing between cramps with identifiable causes and idiopathic cases. For nonidiopathic cases, management consists of treating the underlying cause whenever possible. A nonpharmacologic approach (massaging and stretching) is the recommended first-line treatment for idiopathic cases. Quinine sulfate also appears to offer safe and effective symptom management of idiopathic cases, although its efficacy has not been definitively established in clinical trials. PMID- 11417374 TI - The aging heart. State-of-the-art prevention and management of cardiac disease. AB - The normal aging heart undergoes pathophysiologic changes that over time undermine cardiac structure and function. In older persons, declining cardiac function can be accelerated or exacerbated by chronic diseases such as hypertension or heart failure. The convergence of normal alterations and distinct cardiovascular conditions--disorders of rhythm, disorders of the organ itself, and vascular disease--compound the challenge of clinical management. This task is becoming somewhat less complex as new information emerges from clinical studies seeking more effective approaches to preventing and managing myocardial infarction, coronary artery disease, chronic heart failure, and systolic and diastolic dysfunction. PMID- 11417375 TI - MMSE. Interpreting mental status examination scores in cases of mild dementia. PMID- 11417376 TI - New benefit: cancer screening with colonoscopy. PMID- 11417377 TI - [The tissue distribution of antibiotics in the udder--comparison of the situation in vivo with the isolated perfused bovine udder]. AB - Especially for animal protection reasons, tissue concentrations of intracisternally administered antibiotics in the mammary gland hardly can be determined in the live cow. Therefore, this paper assessed the use of the isolated perfused bovine udder to study the distribution of penicillins in glandular tissue. With this intention, the in vitro results acquired with this model were compared to tissue concentrations as well as absorption data from the few in vivo studies in the literature and differences were interpreted. This approach must consider inevitable deviations of experimental materials and methods. Furthermore, in vivo the udder is included in a closed circulatory system with other metabolism and excretion feasibilities than the isolated model. Moreover, the lower flow rate in the vessels in vitro has to be taken into account with respect to absorption capacities. Nevertheless, the tissue concentrations and the distribution equilibrium across the blood-udder-barrier in both experimental concepts corresponded with each other, if the deviating conditions are considered by calculating correction factors. Advantages and disadvantages of the isolated udder are discussed critically. In conclusion, this method is a useful completion of pharmacokinetic in vivo studies that are supposed to compare intracisternally administered formulations. PMID- 11417378 TI - [Increase in the prevalence of Echinococcus multilocularis infection in red foxes in Lower Saxony]. AB - Echinococcus multilocularis is a tapeworm having carnivores as final hosts, the red fox in particular, dog and cat less frequent. Its two host life cycle consists of a larval cycle which predominantly takes place in the liver of rodents such as mice but it can also develop in musk rats as intermediate hosts. Man can also be infected and serves as a wrong intermediate host. He develops an alveolar echinococcosis which usually ends lethal without medical treatment. The prevalence of E. multilocularis among 5.365 red foxes in Lower Saxony was monitored from 1991 to 1997. The data were analysed using spatial epidemiological methods. This evaluation is based on a significance test which was applied to the parameters of spatial autoregressive regression models (CAR) fitting to the data of two successive sampling periods from 1991-1994 and 1994-1997. The mean prevalence (spatial median) increased from 6% to 11%. The results of this epidemiological study which was restricted to Lower Saxony support for the first time the earlier reported assumption that the prevalence of foxes infected with E. multilocularis has risen in Europe in the last decade. The reasons for this development are still unknown. PMID- 11417379 TI - [Breed and sex differences in the temperament of calves in a cow-calf operation]. AB - Behavioural traits related to temperament were studied in German Angus (Aberdeen Angus x German dual purpose breeds) and German Simmental cattle. Temperament was defined as the animal's behavioural response to handling. 498 calves (279 German Angus, 219 German Simmental) were tested with two different restrain tests. The animals were tested three times individually during a fixation procedure (days of live: 5, 19, 33) or weighting (days of live: 62, 138, 193). LSQ-means were calculated with a model including breed, year and sex as fixed effects and the sire as a random. Furthermore a regression on the age of the calf was included. Behaviour of the animals were recorded. Significant differences in the recorded parameters were found between the breeds on day 19 and 33 for one test and day 62 for another test. German Angus and male calves were less difficult to handle regarding the different parameters. Both tests could be used to show differences between the breeds. PMID- 11417380 TI - [Differential diagnosis of classical swine fever and border disease: seroepidemiological investigation of a pestivirus infection on a mixed sheep and swine farm]. AB - During recent years neutralizing antibodies against Border Disease Virus (BDV) were found repeatedly in German pig herds. Consequently there was a demand for a differential diagnostic system. A permanent sheep cell line and BDV reference strain Moredun were chosen and were applied in a could be used case study. A pestivirus could be isolated from piglets on a mixed farm and was characterised as 'non-Classical Swine Fever' (CSF) by using monoclonal antibodies. Due to a CSF suspicion the pig herd was destroyed immediately. Serum samples of sheep from the same farm were used for further characterisation of the new virus isolate. A neutralization test of the sheep sera was performed against different pestiviruses and the new isolate. Neutralizing antibody titres against the new virus pig isolate were significantly higher than against all other pestiviruses. BDV strain Moredun recognised the antibodies clearly, whereas CSF viral strain Alfort 187 and several isolates of bovine viral diarrhoea virus (BVDV) strains scored the lowest cross reaction. PMID- 11417381 TI - [A modification of the in ovo-method for the determination of the antihemorrhagic activity of plant extracts]. AB - Up to now there has been only a method available for testing the antihaemorrhagic activity of plant extracts, in which chicken embryos had to be transferred to a special container on the fifth day of incubation. Furthermore, the test could not be conducted on the extraembryonal circulatory system until two days later. In the test method described here the embryos remain inside the egg. After removal of the eggshell through the air cell and removal of the inner shell membrane, direct access to the extraembryonal circulatory system is possible, minimising external influences on the test system. This represents a dramatic simplification in technique and permits testing already on the fourth day of incubation. The application of this method to determine the antihaemorrhagic properties of samples has the advantage of permitting the simple and quick production of haemorrhages by snake venom on the yolk sac circulatory system. PMID- 11417382 TI - Ulcerative and papillomatous digital dermatitis of the pastern region in dairy cattle: clinical and histopathological studies. AB - The study was carried out on a single herd of 500 Holstein breed dairy cattle. Only 50 dairy cows (10%) were found to be suffering from focal ulcerative and papillomatous digital dermatitis of the pastern region. The cows were subjected to clinical and histopathological examinations. Early ulcerative stage was seen in 30 cows and late papillomatous stage was observed in 20 cows. The topical application of oxytetracycline solution or benzathine penicillin powder appeared to be effective for the treatment of the ulcerative lesions. The histopathological findings showed signs of epidermopoiesis and papilloma formation. The dermal reaction revealed signs of fibroplasia and perivascular aggregation of inflammatory cells. Silver stained sections indicated the presence of longer filamentous spiral spirochetes, cocci and large-sized anaerobic bacilli invading the stratum spinosum. In conclusion it can be said that bovine digital dermatitis was observed in ulcerative and papillomatous form at the pastern region bordering the dewclaws. Spirochetes are frequently associated with and may be responsible for pathological changes in the lesion. PMID- 11417383 TI - An attempt for complex assessment of the elderly patients' compliance. AB - Compliance is taking the correct amount of the correct medicine at the correct time. Today, the problem of prescription medicine noncompliance is gaining considerably more attention within the pharmacy community as a part of the pharmaceutical care and by the other health professionals. Sometimes, compliance is defined "objectively" by how much medication a patient takes, such as taking a certain percentage of prescribed medication or taking a certain number of pills over a specific time period. Undoubtedly the care for the elderly patients and especially for the elderly patients that are bedridden can be an especially difficult challenge. The extent of non-compliance among them is estimated to be about 40% to 50%. The aim of this research is to assess the level of compliance and the factors, that influence this process among a group of elderly patients. We applied a complex methodology in order to avoid the mistakes that are connected with the use of a single method separately. PMID- 11417384 TI - Analysis of the expenses for drugs in the stationary departments of the united regional hospitals. AB - This issue is an attempt for estimation of the expenses for drugs in a stationary departments of the united regional hospitals in Sofia. We applied variation, unparametric, alternative methods. The results from this issue will improve the health care in Bulgaria and will provide the introduction of the new healthcare reform. PMID- 11417385 TI - Rapid high-performance liquid chromatographic method for determination of doxycycline in human plasma. AB - A simple, rapid and precise analytical method for determination of doxycycline in human plasma is described. A high-performance liquid chromatography (HPLC) system based on a mbondapak C18 column (300 mm x 4.6 mm i.d., 10 microns) and a UV detector (lambda = 347 nm) were used. A mixture of actonitrile 0.1 M potassium dihydrogen phosphate buffer (35:65 v/v) adjusted to pH 3 was used as mobile phase. The proteins were precipitated with a 24% perchloric acid aqueous solution. The detection limit for doxycycline in plasma was 3 ng mL-1. The average recovery was about 96%. The method is simple, sensitive and suitable for pharmacokinetic studies of doxycycline. The inter-day and intra-day assay coefficients of variation were found to be less than 8%. PMID- 11417386 TI - An ion-pair high-performance liquid chromatography method for stability studies of epinephrine acid tartrate in injectable solution. AB - An isocratic technique was developed for the analysis of epinephrine acid tartrate using ion-pair high-performance liquid chromatography (HPLC) with UV detection and a phenyl reversed phase column. The chromatographic separation was accomplished by a mixture mobile phase consisting of methanol: water (50:50, v/v) containing 7.28 mM sodium dodecyl sulfate adjusted to pH 3.5. The stability of the drug in injectable solution was studied. Using the kinetic information generated from the high temperature studies at 70, 80 and 90 degrees C, an Arrhenius plot for epinephrine acid tartrate degradation was constructed and the shelf life of the drug was estimated. We concluded that the shelf life of epinephrine acid tartrate at room temperature (25 degrees C) was 346.1 days. PMID- 11417387 TI - Studies on the oestrogenic activity of soybean oil on albino rats. AB - The oestrogenic activity of soybean oil was studied biologically by vaginal epithelium cornification and the uterine weight method. A significant increase in vaginal cornification was found in ovariectomized adult rats given soybean oil orally for five days resulting an oestrogenic like activity for the soybean oil. Also, using the uterine weight method in immature and ovariectomized rats, the administration of either oestradiol or soybean oil induced a significant increase in the uterine weight. These results indicated that the effect of soybean oil has an oestrogenic like activity equal to 0.6 that activity of oestradiol. Furthermore, with respect to body weight the administration of either oestradiol or soybean oil showed insignificant change than the initial body weight. These findings suggest that soybean oil exerts an oestrogenic like effect. PMID- 11417388 TI - Evaluation of adhesive properties of transdermal therapeutic systems containing nitroglycerin. AB - The patch performances and the success of the transdermal drug delivery can be significantly affected by the quality of contact between the patch and the skin. Poor adhesion will dramatically reduce percutaneous delivery. In this study the adhesive properties (peel force and creep resistance) of three monolayer self adhesive nitroglycerin (NTG) patches available on the market, Deponit, Minitran, and Triniplas, were compared. The patches were characterized also in terms of in vitro drug release and ex vivo skin permeation. The creep resistance values verified in the case of Deponit and Triniplas indicated a low cohesion of these matrices. The peel force values were in the accepted range, even if Triniplas and Deponit showed values double that shown by Minitran. The percentage of NTG released in vitro after two hours in all cases exceeded ninety percent. The ex vivo permeation profiles were similar, even if the three patches had different loaded amounts and surface areas. The measured permeated amount, 11 mg permeated in 24 h, was predictive of the claimed in vivo release (10 mg in 24 h). PMID- 11417389 TI - Cissus stem gum as potential dispersant in pharmaceutical liquid systems 2: The emulsifying and suspending properties. AB - The emulsifying and suspending properties of a new gum derived from the stem of cissus rufescence family Amphelidaceae were studied. Stability of the liquid paraffin emulsions prepared using this mucilaginous substance was compared with that containing tragacanth or acacia. The rate of globule coalescence was determined using Sherman's equation for concentrated emulsions. The suspending ability of the polymer was compared with that of tragacanth or compound tragacanth. The rate of deflocculation, K, was found to obey a power law equation: beta t = beta 0 e-kt in zinc oxide suspensions. At concentrations above 0.75% w/v, cissus gum produced liquid paraffin emulsion with minimal separation. The rate of globule coalescence was in the order acacia > cissus > tragacanth and rate of creaming was tragacanth > acacia > cissus. At concentrations of 0.6 to 1.0% w/v, cissus gum produced highly flocculated zinc oxide suspensions, which exhibited good redispersibility. Stability of the agglomerated, dispersed particles was similar to that produced using tagacanth mucilage. PMID- 11417390 TI - Product development studies for stabilization of oral liquid enzyme preparations containing diastase and papain. AB - A product development study was undertaken with a view to stabilize acid sensitive amylolytic enzyme diastase and alkali sensitive proteolytic enzyme papain. An unique high technology method was adopted for both enzymes with enteric coating and non-enteric film forming materials respectively to achieve site specificity of action, better stability and to protect diastase from the adverse acid pH of the stomach. Feasibility of incorporating various excipients like thickening agents, surfactants, preservatives, etc., to produce stable elegant oral liquid suspension was also studied. Release studies of the enzymes both in simulated gastric and simulated intestinal juices showed steady and consistent release. The proteolytic and amylolytic activities were assessed by standard pharmacopoeial methods. The stability of the product during the studies was satisfactory. Reduction in addition of overages of costly enzymes met the project technically feasible and economically viable. PMID- 11417391 TI - Benzyl ester prodrug of ibuprofen: pharmacological and toxicological profile. AB - Prodrug approach is a promising way of overcoming gastrotoxicity associated with long term oral use of NSAIDs like ibuprofen. The candidature of benzyl ester prodrug of ibuprofen was examined to assess its ability to reduce gastrotoxicity without affecting pharmacological response. It gave highly promising activity profile in established animal models like carrageenan induced rat paw oedema and acetic acid induced writhing reflex assay. The effect of physicochemical properties like aqueous solubility, octanol-water partition coefficient, hydrolysis kinetics in human plasma and simulated gastric fluid, on activity profile has been investigated. PMID- 11417392 TI - Synthesis of 1,2,4-triazino[5,6-b] indoles bearing 1,2,4-triazine moiety. AB - Some new 3-(5,6-Diphenyl-1,2,4-triazin-3-yl)-5-substituted- 1,2,4-triazino[5,6 b]indole derivatives (6-12) have been obtained via treatment of both (4 and 5) with p-nitro-benzoyl chloride, ammonium thiocyante, formaldehyde-methanol, acrylonitrile and thiosemi-carbazide. The former structure of the new products was established by the help of elemental analyses, as well as spectral data. Some of their showed pronounced effect on the Cellobiase produced by Thermomyces lanuginosus and Chaetomium thermophilum. PMID- 11417393 TI - Synthesis of novel 2,4 (1H, 3H)-quinazolinedione derivatives with analgesic and anti-inflammatory activities. AB - Starting from mefenamic acid a series of 1-(2,3-dimethyl-phenyl)-2,4(1H,3H) quinazolinedione derivatives were prepared. Structures of the new compounds were confirmed by microanalysis, IR, and 1H-NMR. Seven representative compounds were subjected to preliminary pharmacological screening which revealed that some of them exhibited analgesic and anti-inflammatory activity greater than mefenamic acid. PMID- 11417394 TI - [New scenarios for research on contrast media for diagnostic imaging]. PMID- 11417395 TI - [Open letter to the Health Minister from a group of epidemiologists and experts in health programming]. PMID- 11417396 TI - [We need to remember the past in order to plan for the future]. PMID- 11417398 TI - [1,200 researchers marching between science and reason]. PMID- 11417397 TI - [Skin cancer day: the debate continues]. PMID- 11417399 TI - [Fiscal federalism and health: how can the health care system be improved?]. PMID- 11417400 TI - [Health models versus election programs]. PMID- 11417401 TI - [Why should the direct participation in the health care expenditure be abolished?]. PMID- 11417402 TI - [The public health agency of Lazio and privacy of AIDS patients]. PMID- 11417403 TI - [Is research untimely in sudden death?]. PMID- 11417404 TI - [Mortality in a cohort of alcoholics from Arezzo in 1979-1997]. AB - This study evaluates the risk of mortality in a cohort of Italian alcoholics resident in a rural area characterised by traditional drinking habits. Individual vital status of the 1,037 patients enrolled at the Centro Alcologico in Arezzo during the 1979-1997 period has been traced. Causes of death (ICD-IX) have been retrieved from the Regional Mortality Register. Standardised mortality ratios (SMR) have been computed according to gender and 5-year age group mortality rates of the general population resident in Tuscany during the same period. This cohort is representative of the traditional rural alcoholism of the Tuscany region based on wine consumption. Among the 9,190 person-years followed-up, 333 deaths have occurred, corresponding to approximately 2.6 fold the expected number of deaths (SMR males: 2.6; females: 2.4). In both genders, significantly high SMRs for liver cirrhosis, oesophagus and liver cancer are reported, while SMRs of cancers at all sites, oral and respiratory cancers, injuries as a whole, road and traffic accidents, and suicides are significantly elevated only among males. No relevant variation between expected and observed deaths for pancreatic diseases, colon cancer, female breast cancer, and, despite a large proportion of heavy smokers, for cardiovascular diseases (hypertension, cerebrovascular diseases, coronary heart diseases) has been recorded. This research confirms the high mortality among a cohort of Italian alcoholics. However, causes of death related with violence and trauma are proportionally less represented, in accordance with the social pattern of Mediterranean alcoholism. The absence of cardiovascular mortality risk in a wine-based cohort of alcoholics is an unexpected finding that requires to be further examined. Finally, to prevent smoking related deaths, alcohol addiction services should begin to introduce smoking cessation practices among treatment protocols. PMID- 11417405 TI - [Occupational exposure to asbestos and recognition of pleural mesothelioma as occupational disease in the province of Genoa]. AB - The present study compares the data of pleural mesothelioma (PM) patients resident in the province of Genoa (Italy) who, in the period 1994-1996, applied to the Italian National Insurance Institute for Work Accident (INAIL), for workers' compensation for asbestos-related diseases due to occupational exposure, with the dataset of PM patients collected by the Mesothelioma Registry of Liguria (REM) in the same period and in the same area. As PM is a malignant tumor of a prevalently occupational origin, it is recognized and acknowledged as such by INAIL when clinical and etiological characteristics are matched. Objectives of this study were to describe observed PM cases, to evaluate completeness of collected data and differences between those who requested compensation and those who did not. The REM describes the incidence of PM among Ligurian residents, proposing patients with a specific questionnaire to assess occupational, environmental and indoor asbestos exposures. The REM collected 199 new cases of PM among the residents of both the city of Genoa (1994-96) and the province of Genoa (1995-96). In the same period, INAIL received 48 (24%) applications for compensation. Among these, 43 subjects were included in a subgroup of 98 patients registered in the REM as cases with definite diagnosis and ascertained asbestos exposure; 32 were awarded compensation, while 11 are under evaluation. The data collected by REM do not show proven asbestos exposure and/or PM diagnosis for five other subjects (two compensated and three under judgment). This study reveals that: a) only a 24% of the patients with a diagnosis of PM and asbestos exposure apply for compensation; b) an exchange of information among institutions involved in primary prevention, in the evaluation of occupational exposures to carcinogens and in insurance compensation is useful. PMID- 11417406 TI - [The epidemiologist in the court: requirements, professional standards, and deontologic considerations]. AB - This paper examines two issues associated with the activity of epidemiologists who act as expert witness in courts: professional standards and deontologic codes. These issues are to be viewed in the light of the various roles the epidemiologist is called to play in court, as consultant for the prosecutor, the defendant and the plaintiffs. The criteria adopted in court should not differ from the ones usually followed in good epidemiologic practice. Potential conflict of interest that may occur when the personal interest of a researcher leads to introduce bias in his/her/witnes is another relevant issue. Another relevant point is how an epidemiologist deals with his/her colleagues in the courts: diversity in opinions and rigorous critics are acceptable but they must be adequately documented. PMID- 11417407 TI - [Inequalities and risk factors: tobacco use]. PMID- 11417408 TI - Regression analysis of multiple outcomes. PMID- 11417409 TI - Microvascular decompression of the facial nerve for hemifacial spasm in youth. AB - OBJECTS: The goal of this study was to investigate the differences between clinical findings in youth and in adulthood on microvascular decompression (MVD) of the facial nerve for the treatment of hemifacial spasm (HFS). METHODS: We retrospectively evaluated 855 patients who underwent MVD from January 1985 to July 1999. In our series of 33 young HFS patients, all patients had definite offending vessels. Interestingly, pathologic tortuous vertebral artery as a possible etiology was more rarely observed in young HFS patients (1/33 patients, 3.0%) than in adult patients (61/822 patients, 7.4%) (P < 0.05). We did not observe any atomical variations of the vessels or any arachnoidal thickening around the root entry zone and cerebellopontine cistern in youths. Furthermore, young HFS patients did not necessarily have poorer surgical outcomes than adult HFS patients. CONCLUSIONS: Our results suggest that the cause and progress of HFS are the same in youth as in adulthood, even though the pathogenesis of early onset remains unclear. PMID- 11417410 TI - Tuberous sclerosis: epileptogenicity and multimodal presurgical evaluations. AB - We report on a patient with tuberous sclerosis-related epilepsy who benefited from surgical treatment. Various presurgical evaluations, including positron emission tomography (PET), made it possible for us to localize the epileptic focus accurately. In this paper, we stress the importance of performing multimodal evaluations to determine which tubers really possess epileptogenicity. In addition, the implications of PET in tuberous sclerosis-related epilepsy are described. PMID- 11417411 TI - Differentiation, proliferation and apoptosis in primary and recurrent primitive neuroectodermal tumors of childhood. AB - Primitive neuroectodermal tumors (PNETs) of the CNS are a group of embryonal tumors composed of small undifferentiated or poorly differentiated cells. Infratentorially located PNETs are a synonym for medulloblastomas. In this study 31 PNETs, including 5 recurrent tumors, were examined. All children underwent neurosurgery and chemotherapy according to the HIT and HIT-SKK protocols. The specimens were investigated both for their expression of nine immunohistochemical markers for neuronal, astrocytic, mesenchymal and epithelial differentiation and for their proliferation. Results regarding cellular differentiation were confirmed ultrastructurally. Apoptosis was detected by labeling the 3'OH ends generated by DNA fragmentation and by electron microscopy. Glial differentiation was shown to have a prognostic relevance, with an elevated (twofold) risk of recurrence. Neuronal differentiation also indicated a tendency to poor prognosis. Those tumors that recurred later showed an increased proliferation rate (69%) compared with nonrecurrent tumors (58%). Apoptosis was identified in all tumors examined. The proportion of apoptotic cells could not be related to the effect of therapy. These results indicate that cellular differentiation may be a useful predicative factor for the prognosis of cerebral PNETs. PMID- 11417412 TI - Advantages of delayed VP shunting in post-haemorrhagic hydrocephalus seen in low birth-weight infants. AB - The optimal timing of ventriculo-peritoneal (VP) shunt insertion in a neonate with post-haemorrhagic hydrocephalus (PHH) is uncertain. During the 8-year period from January 1989 to December 1996, 41 patients had VP shunts inserted for PHH at Red Cross War Memorial Children's Hospital. Data on 36 patients were sufficient for review in order to determine whether the timing of surgery in any way influenced the complication incidence of this intervention. Nine of the 36 patients had a serious complication, either infection or mechanical shunt obstruction, during their initial hospital admission (early period). Nineteen patients had surgery performed before day 35 after birth and all those with early complications were in this group. Seventeen patients had surgery delayed until after day 35, and none of these patients had an early complication. In this study a higher complication incidence was noted when a VP shunt was inserted prior to day 35 (Chi-square test P < 0.01). This most probably correlates with a high concentration of blood breakdown products in the cerebrospinal fluid during the first month after intra-ventricular haemorrhage. PMID- 11417413 TI - Treatment of post-haemorrhage ventricular dilatation with an Ommaya's reservoir: management and outcome of 64 preterm infants. AB - Over a 5-year period, an Ommaya's reservoir has been inserted in a single neurosurgical centre in each of 64 preterm infants with post-haemorrhage ventricular dilatation (PHVD). Their mean gestational age at birth was 29.2 weeks. The average age at reservoir insertion was 24 days. Seventeen infants received a fibrinolytic agent through the reservoir. Infections occurred in 14 patients. Two patients died after handling of their reservoirs. Thirty-one of the 45 survivors required a shunt placement. After a follow-up ranging from 6 months to more than 4 years, 17 of 43 patients have severe sequelae or are handicapped. Compared with the results of other studies, our experience does not suggest that treatment of PHVD with an Ommaya's reservoir is beneficial in term of mortality, shunt placement, and/or neurological outcome. The place of Ommaya's reservoir among the various means of managing PHVD should be carefully evaluated, as should the best way of using this appliance safely. PMID- 11417414 TI - The role of gamma knife radiosurgery in children. AB - OBJECTS: Despite advances in microneurosurgery, the surgical management of deep seated lesions is still associated with a high risk. Gamma knife radiosurgery (GKRS), however, has improved the outcome of cerebral tumors and arteriovenous malformations (AVMs) in delicate areas. METHODS: Between March 1992 and June 1998, 50 children (age 1-16 years) with intracranial lesions were treated with GKRS. There were 12 low-grade gliomas, 12 high-grade gliomas, 7 craniopharyngiomas, 3 hamartomas, 2 meningiomas of the skull base, 2 vestibular schwannomas, 1 pituitary adenoma, 1 choroid plexus papilloma, and 10 AVMs. The mean size of the pathologies was 4.6 cm3 (range: 0.21-25.5 cm3). A mean marginal dose of 16 Gy (8-25 Gy) was applied to a mean isodose surface of 50% (35-90%). Clinical and neuroradiological follow-up were analyzed for outcome. Follow-up periods of 45 of these patients ranged from 8 to 79 months (mean 36 months); 5 patients were lost to follow-up. Neoplasms decreased in size in 15 cases (41%), remained unchanged in 13 patients (35%), and increased in 9 cases (24%). AVMs obliterated in 3 children (38%) within 24 months. Neurological follow-up demonstrated improved clinical status in 7 patients (15.5%), stable neurological status in 31 cases (69%) and clinical deterioration in 7 patients (15.5%). The treatment was well tolerated and no complications occurred. CONCLUSIONS: GKRS represents a safe and effective treatment option for benign neoplasms or AVMs in pediatric patients and may extend survival times of children with malignant lesions. PMID- 11417415 TI - Dandy-Walker syndrome: different modalities of treatment and outcome in 42 cases. AB - OBJECTS: Forty-two patients with Dandy-Walker syndrome who were treated with different surgical modalities over a period of 8 years, from 1988 to 1996, at the Sanjay Gandhi Post Graduate Institute of Medical Sciences were reviewed in the present study. METHODS: All the patients presented with hydrocephalus at the time of diagnosis. Association of other CNS anomalies was detected in 9 (22%) patients. Vermian hypoplasia was present in 36 (88%) cases, while cerebellar hypoplasia was documented in 27 (59%) of these patients. The treatment modality for these cases has continued to change in our institution over the years. Initially the ventriculoperitoneal shunt was the treatment of choice and was performed in 28 (66%) patients. Subsequently shunting of the cyst to the peritoneal cavity was performed in 7 (16%) patients. More recently, 3 of our patients were treated by fenestration of the cyst membrane and 4 others (9%), by ventriculocystoperitoneal shunting procedures. In this study the additional insertion of cystoperitoneal shunts was required in 8 (27%) of the 28 patients who had primary ventriculoperitoneal shunt procedures, because of either nonresolution or reappearance of a posterior fossa cyst. Six out of the 7 cases of primary cystoperitoneal shunts required additional ventriculoperitoneal shunt diversion because of persistent ventricular dilatation. In the group of 4 patients with primary ventriculocystoperitoneal shunts, only 1 patient required shunt revision. The patients on whom cyst membrane fenestration was performed required no additional procedures, except for 1 who already had a ventriculoperitoneal shunt in situ. CONCLUSIONS: Therefore, 18 of the total 42 patients could ultimately be made asymptomatic by ventriculocystoperitoneal shunting for one or the other of these reasons. PMID- 11417416 TI - Congenital dermoid cyst of the anterior fontanel in mestizo-mulatto children. AB - OBJECTS: Twenty-seven cases of histologically confirmed congenital dermoid cysts of the anterior fontanel in children are reported. METHODS: The age, sex and race of each patient was recorded. CONCLUSIONS: Ages ranged between 2 months and 6 years. There was a female predominance, and 77.7% of these patients were children of mixed race. Surgical excision resulted in complete cure without complications or recurrences. PMID- 11417417 TI - Cortical blindness: an unusual complication after removal of a ganglioneuroma of the neck. AB - A 5-year-old girl was operated upon after a huge mass had been found on the left side of her neck. Preoperatively the anatomy was studied using MRI, including 3D reconstruction of the tumor. The left vertebral artery was seen to cross the tumor, and during the operation the vessel was preserved. Postoperatively it thrombosed and a permanent cortical blindness developed. One year later MRI angiography of the intracerebral vessels was performed and a pre-existing anomaly of the circle of Willis was demonstrated, which had contributed to the development of the cortical blindness. PMID- 11417418 TI - Pott's puffy tumour: still not an eradicated entity. AB - Pott's puffy tumour is an infrequent entity characterised by one or more subperiosteal abscesses associated with frontal bone osteomyelitis. Although cases in patients of all ages have been reported, teenagers are the most frequently affected. Early diagnosis and aggressive treatment are essential because of the high risk of severe neurological complications, such as epidural abscess, subdural empyema, and secondary septic thrombosis of the dural sinuses. This paper describes the case of a patient with a subperiosteal abscess resulting from sinusitis, with orbital and intracranial extension, and subsequent neurological complications. Despite modern methods of diagnosis and treatment, 13 new cases have been published in the last 5 years; in at least 3 (23%) of these cases there were serious neurological complications. Upper respiratory infections and sinusitis are leading causes of visits to the emergency department in the paediatric age group; however, no risk factors for poor outcome have so far been identified in any of these patients. PMID- 11417419 TI - Lateral recess cysts in two cases of isolated fourth ventricle. AB - We report two cases of isolated fourth ventricle with lateral recess cysts. The background, clinical features, treatment and outcomes are presented and discussed. We speculate on the association of these two conditions and propose possible explanations for the clinical features. PMID- 11417420 TI - Bilateral internuclear ophthalmoplegia following mild head injury. AB - A 7-year-old child presented with bilateral internuclear ophthalmoplegia (INO) following a trivial head injury. CT was normal. MRI revealed a pontine lesion. Two months after the injury the patient was neurologically normal. INO following head injury is rare. Rarer still is INO following mild head injury. To date, only four cases of INO had been reported following mild head injury; the present case is the fifth and the first in which the lesion was documented using MRI. The relevant literature is reviewed. PMID- 11417421 TI - Possible role of adjunctive thalidomide therapy in the resolution of a massive intracranial tuberculous abscess. AB - We present the case of a young child who developed a massive tuberculous abscess of the posterior fossa while being treated for pulmonary tuberculosis. Clinical improvement after surgical excision of the abscess was followed by recurrence of symptoms of acutely raised intracranial pressure on standard antituberculosis and corticosteroid therapy. Magnetic resonance (MR) imaging of the brain showed that a multiloculated abscess had developed anterior to the excision site of the original abscess. The recurring abscess was partly excised and drained but could not be removed completely because of its proximity to the brain stem. Thalidomide, a potent inhibitor of tumour necrosis factor alpha (TNF-alpha), was added to the treatment regimen and resulted in marked clinical improvement with resolution of the abscess within 4 months. The remaining CT lesion had the appearance of a small granuloma. Both the clinical and the radiological response was maintained after 1 year of antituberculosis treatment. PMID- 11417422 TI - Cervical encephalocele in a newborn--Chiari III malformation. Case report and review of the literature. AB - Chiari malformations are brain stem anomalies with or without spinal canal involvement. The combination of brain stem and cerebellar anomaly with a cervical cele is named Chiari III malformation. Patients with this extremely rare defect frequently present with severe neurological defects and can have a poor prognosis. Following a report on a newborn child with a Chiari III malformation that took a fatal but typical course the literature is reviewed. PMID- 11417423 TI - Analysing entrainment of cardiac and locomotor rhythms in humans using the surrogate data technique. AB - Using the surrogate data technique we evaluated whether, during running, the synchronization between cardiac and locomotor rhythms resulted from entrainment or by chance. An electrocardiogram and an electromyogram from the right vastus lateralis muscle were monitored from ten healthy young men running at a paced rhythm of 150 steps a minute. The relationship between cardiac and locomotor rhythms was determined by examination of the occurrence of the heart beat with respect to the locomotor phase. The examination revealed that synchronization patterns were observed in all subjects. We generated surrogate data by sorting randomly the original locomotor rhythm, and no synchronization patterns were then seen. This may indicate that the synchronization between the cardiac and locomotor rhythms represented entrainment. We have provided the first evidence for the rejection of the hypothesis that when heart beat rhythm is close to the locomotor rhythm, synchronization between the two rhythms occurs by chance. PMID- 11417424 TI - Cardiopulmonary and metabolic responses in healthy elderly humans during a 1-week hiking programme at high altitude. AB - Worldwide there are approximately 100 million visitors to high altitude annually and about 15% of those are elderly. Nevertheless, basic information on the cardiopulmonary and metabolic responses to physical activity at high altitude in the elderly is scarce. Therefore, we studied 20 voluntary healthy elderly subjects (55-77 years) who were randomly assigned to a low- (600 m) or a high altitude (2,000 m) group. Both groups increased the duration of their daily hiking from 2.5 to 5 h during a period of 1 week. Pre- and post-hiking cardiopulmonary variables at rest were measured daily. Exercise tests (3 min step test) were performed on days 1, 4 and 7. Of the morning values at rest, only arterial oxygen saturation (SaO2) had decreased after the 1st night at high altitude. After hiking however, SaO2 was diminished on all days at high altitude. Post-hiking heart rates increased from baseline on days 1 and 2 in the low- and on days 1-5 in the high-altitude group. Exercising SaO2 (%) in the three tests was decreased [84.9 (SD 2.8), 88.1 (SD 2.1), 87.2 (SD 2.3)] compared to baseline [93.2 (SD 2.0); P < 0.05] and blood lactate concentrations were increased [3.1 (SD 0.7), 3.4 (SD 0.3), 3.3 (SD 0.2)] compared to baseline [2.7 (SD 0.6); P < 0.05] in all tests at high altitude. The 1-week hiking programme was well tolerated by the healthy elderly at both low and high altitudes. Ventilatory adaptation to high altitude in the elderly seemed to have been completed within the first 2 days during the measurements at rest. However, cardiopulmonary and metabolic responses to exercise were increased and recovery from exercise was delayed during the 1-week hiking programme at high altitude. Heart rate and SaO2 measurements are considered to be highly sensitive in estimating the state of acclimatisation and for monitoring exercise intensity and duration at high altitude. PMID- 11417425 TI - Calf and shin muscle oxygenation patterns and femoral artery blood flow during dynamic plantar flexion exercise in humans. AB - The effects of dynamic plantar flexion exercise [40, 60, and 80 contractions.min 1 (cpm)] on calf and shin muscle oxygenation patterns and common femoral artery blood flow (Qfa) were examined in six female subjects [mean age 21 (SD 1) years] who exercised for 1 min at 33% of their maximal voluntary contraction at ankle angles between 90 degrees and 100 degrees. Spatially resolved near-infrared spectroscopy was used to measure medial gastrocnemius, lateral soleus (synergist) and anterior tibialis (antagonist) muscle oxygen saturation (SO2, %). Qfa was measured by ultrasound Doppler. The SO2 changed significantly only in the medial gastrocnemius and its decrease (up to about 30%) was independent of the contraction frequencies examined. The increase in Qfa, at the end of exercise, was highest at 80 cpm. When the exercise at 60 cpm was prolonged until exhaustion [mean 2.7 (SD 1.1) min], medial gastrocnemius SO2 decreased, reaching its minimal value [mean 30 (SD 10)%] within the 1st min, and had partially recovered before the end of the exercise with concomitant increases in total haemoglobin content and Qfa. These results suggest that the medial gastrocnemius is the muscle mostly involved in dynamic plantar flexion exercise and its oxygen demand with increases in contraction frequency and duration is associated with an up-stream increase in Qfa. PMID- 11417426 TI - A 15-year physical activity pattern is positively related to aerobic fitness in young males and females (13-27 years). AB - The purpose of this paper is to test the hypothesis that daily physical activity over a period of 15 years has been beneficial to aerobic fitness in young male and female participants (13-27 years) in the Amsterdam Growth and Health Longitudinal Study. Only subjects with the maximal data of six sets of measurements were included (83 male and 98 female participants). Daily physical activity was assessed using a standardized interview on activity and expressed as a weighted activity score. Aerobic fitness was assessed using a maximal running test on a treadmill and measuring the maximal oxygen uptake (VO2max) and the maximal slope of the track (Smax). To assess the longitudinal relationship between daily physical activity and aerobic fitness a real longitudinal analysis was carried out with generalized estimating equations, adjusting for differences in initial aerobic fitness at age 13, and for other lifestyle (dietary intake, smoking and alcohol consumption) and biological parameters (biological age, body fat, blood pressure and concentration of serum cholesterol). A significant relationship (P < 0.01) was observed between daily physical activity and both VO2max and Smax. It can be concluded that the development of aerobic fitness between the age of 13 and 27 years is independently and positively related to daily physical activity in this group of male and female participants in the study. The functional implications, however, are small: a relatively high increase in the weighted physical activity score of 30% over a period of 15 years results in a 2%-5% increase in aerobic fitness. PMID- 11417427 TI - Blood lactate exchange and removal abilities after relative high-intensity exercise: effects of training in normoxia and hypoxia. AB - The effects of 4 weeks of endurance training in conditions of normoxia or hypoxia on muscle characteristics and blood lactate responses after a 5-min constant-load exercise (CLE) at 90% of the power corresponding to the maximal oxygen uptake were examined at sea-level in 13 sedentary subjects. Five subjects trained in normobaric hypoxia (HT group, fraction of oxygen in inspired gas = 13.2%), and eight subjects trained in normoxia at the same relative work rates (NT group). The blood lactate recovery curves from the CLE were fitted to a biexponential time function: La(t) = La(0) + A1(1 - e- gamma 1.t) + A2(1 - e- gamma 2.t), where the velocity constants gamma 1 and gamma 2 denote the lactate exchange and removal abilities, respectively, A1 and A2 are concentration parameters that describe the amplitudes of concentration variations in the space represented by the arterial blood, La(t) is the lactate concentration at time t, and La(0) is the lactate concentration at the beginning of recovery from CLE. Before training, the two groups displayed the same muscle characteristics, blood lactate kinetics after CLE, and gamma 1 and gamma 2 values. Training modified their muscle characteristics, blood lactate kinetics and the parameters of the fits in the same direction, and proportions among the HT and the NT subjects. Endurance training increased significantly the capillary density (by 31%), citrate synthase activity (by 48%) and H isozyme proportion of lactate dehydrogenase (by 24%), and gamma 1 (by 68%) and gamma 2 (by 47%) values. It was concluded that (1) endurance training improves the lactate exchange and removal abilities estimated during recovery from exercises performed at the same relative work rate, and (2) training in normobaric hypoxia results in similar effects on lactate exchange and removal abilities to training in normoxia performed at the same relative work rates. These results, which were obtained non-invasively in vivo in humans during recovery from CLE, are comparable to those obtained in vitro or by invasive methods during exercise and subsequent recovery. PMID- 11417428 TI - Determinants of maximal cycling power: crank length, pedaling rate and pedal speed. AB - The purpose of this investigation was to determine the effects of cycle crank length on maximum cycling power, optimal pedaling rate, and optimal pedal speed, and to determine the optimal crank length to leg length ratio for maximal power production. Trained cyclists (n = 16) performed maximal inertial load cycle ergometry using crank lengths of 120, 145, 170, 195, and 220 mm. Maximum power ranged from a low of 1149 (20) W for the 220-mm cranks to a high of 1194 (21) W for the 145-mm cranks. Power produced with the 145- and 170-mm cranks was significantly (P < 0.05) greater than that produced with the 120- and 220-mm cranks. The optimal pedaling rate decreased significantly with increasing crank length, from 136 rpm for the 120-mm cranks to 110 rpm for the 220-mm cranks. Conversely, optimal pedal speed increased significantly with increasing crank length, from 1.71 m/s for the 120-mm cranks to 2.53 m/s for the 220-mm cranks. The crank length to leg length and crank length to tibia length ratios accounted for 20.5% and 21.1% of the variability in maximum power, respectively. The optimal crank length was 20% of leg length or 41% of tibia length. These data suggest that pedal speed (which constrains muscle shortening velocity) and pedaling rate (which affects muscle excitation state) exert distinct effects that influence muscular power during cycling. Even though maximum cycling power was significantly affected by crank length, use of the standard 170-mm length cranks should not substantially compromise maximum power in most adults. PMID- 11417429 TI - Sodium replacement and fluid shifts during prolonged exercise in humans. AB - In the study presented here, we examined the affects of a close to complete replacement of sweat water and Na+ losses on fluid shifts during exercise. Six cyclists performed three 4-h rides at 55% of their peak oxygen uptake in a 20 degrees C environment while consuming 3.85 l of an 8% carbohydrate solution containing 5, 50 or 100 mEq.l-1 of Na+. Increases in Na+ intake reduced renal free water clearance from around 40 ml.h-1 to -8 and -121 ml.h-1 and led to a decrease in urine volume from approximately equal to 1.0 to 0.5 l (P < 0.05). In contrast, the 3.5-3.9 l fluid and 150-190 mEq Na+ losses in sweat were similar in each trial, as were the approximately equal to 80 mEq K+ losses in sweat and urine and the 282-288 mosmol.kg-1 plasma osmolalities. During the low-Na+ trial, plasma osmolality was maintained by a approximately equal to 1.3 l contraction of extracellular fluid (ECF) with the loss of approximately equal to 200 mEq Na+. However, in the other trials, approximately equal to 1.3 l of water was lost from the intracellular fluid. During the medium-Na+ trial, a loss of only approximately equal to 40 mEq Na+ maintained ECF volume, and during the high-Na+ trial, a gain of approximately equal to 160 mEq Na+ expanded the ECF by approximately equal to 0.8 l. However, corresponding changes in plasma volumes from -0.20 to 0.15 l had no effect on cardiovascular drift or thermoregulation. These data suggest that during prolonged exercise of moderate intensity under mild environmental conditions when sweat rates are approximately equal to 0.9 l.h 1, complete Na+ replacement maintains plasma volume and reduces dehydration, but when fluid intake matches sweat rate, has little effect on plasma osmolality. PMID- 11417430 TI - The effects of supplementation with 19-nor-4-androstene-3,17-dione and 19-nor-4 androstene-3,17-diol on body composition and athletic performance in previously weight-trained male athletes. AB - The purpose of this study was to determine the effects of 8 weeks of norsteroid supplementation on body composition and athletic performance in previously weight trained males. Subjects were weight and percent body fat matched and randomly assigned to receive either 100 mg of 19-nor-4-androstene-3,17-dione (N-dione) and 56 mg of 19-nor-4-androstene-3,17-diol (N-diol; 156 mg total norsteroid per day), or a placebo (a multivitamin). Each subject participated in resistance training 4 days/week for the duration of the study. Body composition was assessed via dual energy X-ray absorptiometry. Circumference measures were taken of a relaxed and flexed arm (maximum circumference of the arm), waist (level of umbilicus), and thigh (15 cm proximal to the patella). Strength was determined with a one repetition maximum bench press, while force and power were determined with a dumbbell bench press (60% body weight) on a Stratec Galileo force platform. Profile of mood states scores were evaluated for vigor and fatigue. There were no significant changes in any of the parameters measured. In conclusion, low-dose supplementation with N-dione and N-diol does not appear to alter body composition, exercise performance, or mood states. PMID- 11417431 TI - Blood flow in the triceps brachii muscle in humans during sustained submaximal isometric contractions. AB - The main purpose of this study was to determine the extent to which blood flow through the profunda artery within the triceps brachii muscle may be compromised during maintained low-force isometric fatiguing contractions. Doppler ultrasound techniques were used to record mean blood velocity and arterial diameter of the profunda brachii artery during sustained isometric contractions of 20% maximal voluntary contraction. The arterial diameter did not change throughout the contraction. Thus, blood velocity was considered to be an indicator of blood flow. The mean blood velocity increased initially and then remained constant during the contraction period. When compared to rest [0.06 (SD 0.03) m s-1] mean blood velocity was significantly larger at the start of the contraction [0.13 (SD 0.07) m s-1] and larger yet during recovery following the contraction [0.30 (SD 0.14) m s-1]. Although blood flow through the conduit artery did not drop during the contraction, the post-contraction hyperaemia suggested that circulatory compromise might have occurred at the level of the capillary beds. PMID- 11417432 TI - Are variations in running economy in humans associated with ground reaction force characteristics? AB - It was hypothesized that certain ground reaction force (GRF) variables are positively correlated with running economy (RE; the aerobic demand at a single speed of running). Excessive momentum changes, quantified by linear impulse measures, as well the free moment applied to the running surface could be considered potentially wasteful efforts in terms of metabolic energy requirements. Recreational runners (n = 16) ran on a treadmill at 3.35 m.s-1 for physiological measurements and overground for biomechanical measurements. Correlation coefficients were calculated between RE and total vertical impulse (TVI), net impulses in three orthogonal directions, and descriptors of the free moment. The TVI and the net vertical impulse were the only GRF characteristics significantly correlated to RE (r = 0.62, r = 0.60, respectively). Greater overall muscle support requirements during ground contact, as represented by TVI, may have been responsible for greater aerobic demand. PMID- 11417433 TI - Resynthesis of muscle glycogen after soccer specific performance examined by 13C magnetic resonance spectroscopy in elite players. AB - The purpose of this study was to examine using 13C-magnetic resonance spectroscopy whether muscle glycogen (Gly) utilized during a simulation of a fatiguing soccer match followed by repeated sprints would be resynthesized during the next 24 h while players consumed their habitual diet. A group of 12 elite young players [mean age 17.5 (SD 0.8) years, mean body mass 68.9 (SD 6.6) kg, mean height 177.0 (SD 5.4) cm] participated in the study. Average muscle Gly content before the simulation was 134 (SD 16) mmol.(kg wet mass)-1 and decreased during the test (P < 0.001) to 80 (SD 29) mmol.(kg wet mass)-1. The value had increased (P < 0.01) to 122 (SD 33) mmol.(kg wet mass)-1 24 h later but it was not significantly different from the value obtained before the soccer test. Dietary analysis of the food intake during the 24 h after the running test revealed that players consumed an average of 2,681 (SD 970) kcal.day-1. Mean daily protein, fat, and carbohydrate (CHO) intakes were 85 (SD 29), 99 (SD 44), and 327 (SD 116) g, respectively. The mean amounts of CHO intake normalised to body mass were 4.8 (SD 1.8) g.(kg body mass)-1. In conclusion, the results of this study showed that despite a CHO intake of less than 5 g.(kg body mass)-1 the habitual diet of soccer players might be sufficient to replenish in 24 h the muscle Gly utilized during soccer specific performance. However, cumulative deficits of about 10% in Gly replenishment as found in the present study might provoke decrements in performance. Thus, players should pay attention to their habitual diets and add more carbohydrates to replenish their daily deficits and perhaps increase their basal levels of intake. PMID- 11417434 TI - A non-invasive measure of changes in blood flow in the human anterior tibial muscle. AB - We used photoplethysmography (PPG) to monitor blood flow changes in the human anterior tibial muscle during arterial occlusion and during isometric and concentric contractions. Single-fibre laser-Doppler flowmetry (LDF) was used as a reference in 12 healthy subjects (5 men, 7 women; mean age 24 years). Post exercise hyperaemic muscle blood flow (MBF) was measured immediately after isometric dorsiflexion of the ankle joint at maximal contraction for 1 min and full range-of-motion dorsiflexion and plantar flexion of the ankle joint for 1 min. A thigh tourniquet was applied for the evaluation of post-occlusive reactive hyperaemia. The MBF (baseline = 100%) was [mean (SD)] 150 (31)% (P = 0.003) by PPG (880 nm) and 182 (66)% (P = 0.012) by LDF. After 1 min of maximal isometric contraction, MBF increased to 150 (51)% (P = 0.003) by PPG (880 nm) and to 169 (43)% (P = 0.005) by LDF. After 1 min of maximal concentric contractions, MBF increased to 158 (59)% (P = 0.003) by PPG (880 nm) and to 170 (99)% (P = 0.008) by LDF. Skin blood flow, PPG (560 nm), did not change significantly after isometric or concentric contractions. The results indicate that reactive hyperaemia after exercise and arterial occlusion can be assessed in the human anterior tibial muscle using PPG. PMID- 11417435 TI - Muscular co-operation during joint stabilisation, as reflected by EMG. AB - The experiment that was carried out consisted of subjects pushing an external object (a heavy pendulum) using stable and unstable handles of increasing mobility. Using this protocol it was possible to distinguish between the motor and stabilising functions of the muscles of the upper extremity. The motor functions were realised by the extensors of the upper extremity, whereas stabilising functions were effected by the muscles spanning the wrist joint. The experiment involved synchronised measurements of the electromyographic (EMG) activity of the muscles in question together with several mechanical quantities revealed against the external object: force, velocity and power. As a result, the instantaneous and global EMG contributions of the extensor and stabilising muscles were determined. It was found that it is the equilibrium state of the object being set in motion and not its mobility (expressed in terms of the number of degrees of freedom) that influences the forces produced by individual muscles. We also suggest that the realisation of stabilising functions by skeletal muscles is a necessary condition of performing any voluntary and co-ordinated movement. PMID- 11417436 TI - The reproducibility of tolerance to lower-body negative pressure and its quantification. AB - The reproducibility of tolerance to lower-body negative pressure (LBNP) has not been assessed sufficiently. Furthermore, there has been confusion concerning the most appropriate index by which LBNP tolerance can be quantified. The purpose of this study was to assess the degree of reproducibility in presyncopal-symptom limited LBNP (LBNPtol), using an LBNP chamber. Twenty physically active subjects [median age (range) 21 (18-27) years] underwent three successive LBNPtol tests with 72-120 h between each test. LBNPtol was quantified using the LBNP tolerance index (LTI; delta mmHg.min), cumulative stress index (CSI; mmHg.min), duration of negative pressure (DNP) and maximum magnitude of negative pressure (MNP). Heart rate (fc), systolic (SBP) and diastolic (DBP) blood pressures from the three repeated tests were compared during a control period. The changes from control to maximum response (fc, SBP, DBP) during LBNP were also compared, and percentage changes in estimated blood volume were measured. There were no statistical differences between any of these comparisons (P > 0.05). LTI and CSI were greater in the third test when compared to the first two tests (P < 0.05). The values for DNP and MNP were not statistically different between tests (P > 0.05). Measures of LTI and CSI showed an acceptable level of reproducibility for the first two repeated tests. However, there was an increase in LBNPtol on the third successive exposure to LBNP. These findings have shown that it is possible to achieve reproducible measures of tolerance to LBNP when using a custom-built chamber. This only applies to a test-retest procedure. Furthermore, these data also suggest that DNP and MNP do not adequately reflect the differences shown in LBNP tolerance when using LTI and CSI as measures. PMID- 11417437 TI - The effect of mathematical modeling on critical velocity. AB - The purpose of this investigation was to examine the effects of mathematical modeling on critical velocity (CV) estimates and the oxygen consumption (VO2), heart rate (HR), and plasma lactate values that corresponded to the five CV estimates. Ten male subjects performed a maximal, incremental treadmill test to determine maximal VO2, and four randomly ordered treadmill runs for the estimation of CV. Two linear, two nonlinear, and one exponential mathematical models were used to estimate CV. Regression analyses were used to determine the VO2, HR, and plasma lactate values that corresponded to the five CV estimates from the relationships for VO2, HR, and plasma lactate versus running velocity from the maximal, incremental test. The nonlinear, three-component model (Nonlinear-3) resulted in a mean CV that was significantly (P < 0.05) less than the mean values derived from the other four models, and was the lowest CV estimate for each subject. The percent of maximal VO2, HR, and plasma lactate values that corresponded to the Nonlinear-3 model were 89%, 93%, and 63%, respectively. These findings indicate that CV estimates differ by as much as 20% depending upon the model used to determine the characteristics of the velocity/time relationship. Future studies are needed to determine which model provides the most valid estimate of the demarcation point between heavy and severe exercise. PMID- 11417438 TI - Anaerobic cycling performance characteristics in prepubescent, adolescent and young adult females. AB - The purpose of this study was to determine whether the relationships between short-term power and body dimensions in young females were similar whatever the age of the individuals. A cohort of 189 prepubescent (mean age 9.5 years), adolescent (mean age 14.4 years) and young adult (mean age 18.2 years) females performed three all-out sprints on a friction-loaded cycle ergometer against three braking forces corresponding to applied loads of 25, 50 and 75 g.kg-1 body mass (BM). For each sprint, peak power including flywheel inertia was calculated. Results showed that a braking load of 75 g.kg-1 BM was too high for prepubescent and adolescent girls. Therefore, when measuring short-term cycling performance in heterogeneous female populations, a braking load of 50 g.kg-1 BM (0.495 N.kg-1 BM) is recommended. During growth, cycling peak power (CPP; defined as the highest peak power obtained during the three sprints) increased, as did total BM, fat-free mass (FFM) and lean leg volume (LLV) (P < 0.001). Analysis of covariance revealed that the slopes of the linear relationships between CPP and biometric characteristics were similar in the three groups (P > 0.7 for the CPP/BM and CPP/FFM relationships, and P > 0.2 for the CPP/LLV relationship). However, the adjusted means were always significantly higher in young women (P < 0.001) compared with both of the other groups. Although differences in performance during anaerobic cycling in growing females are primarily dependent upon body dimensions, other as yet undetermined factors may be involved during late adolescence. PMID- 11417439 TI - Muscle enzyme adaptation to training and tapering-off in spinal-cord-injured humans. AB - The activity of muscle metabolic enzymes depends on the amount and type of physical training. We examined muscle enzyme adaptation to prolonged training followed by a period of lowered activity in spinal-cord-injured individuals (SCI). Ten SCI [mean age 35 (SEM 2) years, mean body mass 78 (SEM 4) kg, mean time post-injury 12 (SEM 2) years and range of lesion C5-T4] were given 12 months of functional electrical stimulation of an upright cycling motion for 30 min a day, three times a week, followed by 6 months of training once a week. Activities of glycolytic (hexokinase HK, lactate dehydrogenase LDH) and oxidative (citrate synthase CS, 3-hydroxyacyl-CoA dehydrogenase HAD) enzymes were determined in biopsies of the vastus lateralis muscle taken at 0, 3, 6, 12, and 18 months of training. The degree of sympathoadrenergic activity was evaluated from arterial concentrations of catecholamines in response to acute exercise. Training three times a week induced increases (P < 0.05) in HK (150%), LDH (40%), CS (100%), and HAD (70%) activities that reached a plateau after 3 months. Peak oxygen uptake and power output during exercise by electrical stimulation rose continuously over the first 12 months. After reducing the amount of training by two-thirds, HK, LDH and CS activities remained elevated above basal levels (P < 0.05), whereas HAD, power output and maximal oxygen uptake returned to pretraining levels (P > 0.05). It is concluded that most improvements in glycolytic and mitochondrial oxidative enzyme activities induced by long-term training can be maintained in spinal-cord injured individuals despite a marked reduction in training frequency unrelated to performance or to the degree of sympathoadrenergic impairment. PMID- 11417440 TI - The effect of acarbose on the pharmacokinetics of rosiglitazone. AB - OBJECTIVES: To investigate whether treatment with acarbose alters the pharmacokinetics (PK) of coadministered rosiglitazone. METHODS: Sixteen healthy volunteers (24-59-years old) received a single 8-mg dose of rosiglitazone on day 1, followed by 7 days of repeat dosing with acarbose [100 mg three times daily (t.i.d.) with meals]. On the last day of acarbose t.i.d. dosing (day 8), a single dose of rosiglitazone was given with the morning dose of acarbose. PK profiles following rosiglitazone dosing on days 1 and 8 were compared, and point estimates (PE) and associated 95% confidence intervals (CI) were calculated. RESULTS: Rosiglitazone absorption [as measured with peak plasma concentration (Cmax) and time to peak concentration (Tmax)] was unaffected by acarbose. The area under the concentration-time curve from time zero to infinity [AUC(0-infinity)] was on average 12% lower (95% CI-21%, -2%) during rosiglitazone + acarbose coadministration and was accompanied by an approximate 1-h (23%) reduction in terminal elimination half-life t1/2 (4.9 h versus 3.8 h). This small decrease in AUC(0-infinity) appears to be due to an alteration in systemic clearance of rosiglitazone and not changes in absorption. These observed changes in AUC(0 infinity) and t1/2 are not likely to be clinically relevant. Coadministration of rosiglitazone and acarbose was well tolerated. CONCLUSION: Acarbose administered at therapeutic doses has a small, but clinically insignificant, effect on rosiglitazone pharmacokinetics. PMID- 11417441 TI - Oral quinine pharmacokinetics and dietary salt intake. AB - OBJECTIVES: The objective was to determine whether or not dietary salt intake affects the relative bioavailability of oral quinine. Salt intake has been shown to alter quinidine bioavailability. METHODS: The pharmacokinetic properties of oral quinine sulphate (600 mg salt) were investigated in seven healthy Caucasian volunteers, in a randomised, crossover study, on low- and high-salt diets. Plasma quinine concentrations were measured by high-performance liquid chromatography (HPLC) and the 24-h urinary sodium excretion was assayed. RESULTS: Although the 24-h urine sodium excretion was significantly higher when the volunteers were on a high-salt diet, there were no significant differences in quinine AUC0-infinity, tmax, and Cmax after the two diets. The median (range) quinine elimination half life was significantly shorter after a high-salt diet [8.5 (4.3-10.2) h] than after a low-salt diet [10.0 (7.6-14.8) h] (P = 0.04). CONCLUSION: Dietary salt does not affect the relative oral bioavailability of quinine sulphate. PMID- 11417442 TI - The interaction of saquinavir (soft gelatin capsule) with ketoconazole, erythromycin and rifampicin: comparison of the effect in healthy volunteers and in HIV-infected patients. AB - OBJECTIVE: The aim of this study was to compare the effect of ketoconazole, erythromycin and rifampicin on the pharmacokinetics of saquinavir soft-gelatin formulation (Fortovase; FTV) in healthy volunteers with that in HIV-infected patients at steady state after administration of 1200 mg three times daily. METHODS: In two open-labelled, randomised, crossover studies pharmacokinetic parameters were calculated in healthy volunteers who received on one occasion multiple doses of 1200 mg FTV three times daily alone and on the other occasion in combination with multiple doses of either 400 mg ketoconazole once daily or 600 mg rifampicin once daily. In another open-labelled, multicentre study, 33 HIV infected patients underwent a pharmacokinetic assessment after 36-51 weeks of treatment with FTV and were then given additionally multiple doses of either 200 mg ketoconazole once daily, 250 mg erythromycin four times daily or 600 mg rifampicin once daily. Pharmacokinetic parameters of saquinavir were determined again at the end of the combination treatment. RESULTS: In healthy volunteers, coadministration of ketoconazole increased saquinavir area under the curve from time 0 to 8 h (AUC0-8 h) by 190% (95% CI: 90-343) whereas coadministration with rifampicin resulted in a decrease for AUC0-8 h by 70% (95% CI: 50-82). In HIV infected patients, coadministration of ketoconazole and erythromycin increased AUC0-8 h of saquinavir by 69% (95% CI: 14-150) and 99% (95% CI: 33-198), respectively. When saquinavir was given together with rifampicin, exposure of saquinavir in terms of AUC0-8 h was decreased by 46% (95% CI: 18-65) compared with the baseline assessment. CONCLUSION: Interactions of saquinavir with ketoconazole, erythromycin and rifampicin were observed in healthy volunteers as well as patients. The effects observed in patients, however, appear to be less pronounced. The enzyme induction caused by rifampicin might lead to subtherapeutic levels of saquinavir and this finding appears to be of clinical relevance. PMID- 11417443 TI - Duplication of CYP2D6 predicts high clearance of desipramine but high clearance does not predict duplication of CYP2D6. AB - OBJECTIVE: Duplication of CYP2D6 causes very rapid metabolism of CYP2D6 substrates such as desipramine. However, we have previously shown that in the Danish population, only about 15% of very rapid metabolisers, defined as subjects with a metabolic ratio of sparteine of 0.15 or less, carried a duplicated allele. The question is whether gene duplication is a relatively rare cause (perhaps predictor) of very rapid metabolism or whether a low metabolic ratio is a poor predictor of this. METHODS: After measuring metabolic ratios anew, we selected six volunteers with duplication of CYP2D6 and metabolic ratios ranging from 0.07 to 0.17 and six volunteers without duplication with metabolic ratios ranging from 0.08 to 0.21. Each subject took 100 mg of desipramine. Blood and urine were collected for 48 h. RESULTS: The median total oral clearance of desipramine was 372 l/h and 196 l/h [median difference 108 l/h (95.9% c.i., -304-598 l/h)] and the median partial clearance of desipramine by 2-hydroxylation was 155 l/h and 87 l/h [median difference 47 l/h (95.9% c.i., -124-141 l/h)] for the group with duplication and the group without duplication, respectively. CONCLUSION: The predictive value of duplication of CYP2D6 is poor; there must be other causes (or predictors) of very rapid metabolism and with much higher frequency than duplication of CYP2D6. PMID- 11417444 TI - Human thiopurine S-methyltransferase activity in uremia and after renal transplantation. AB - In addition to cyclosporin and steroids, azathioprine is frequently used for immunosuppression after renal transplantation. Thiopurine S-methyltransferase (TPMT) catalyses the S-methylation of thiopurine drugs. A genetic polymorphism was shown with 1 in 300 homozygous for a TPMT deficiency. These subjects carry the risk of severe myelosuppression when treated with azathioprine. OBJECTIVE: To investigate the influence of hemodialysis on TPMP activity in uremic patients and the effect of azathioprine treatment on enzyme activity. METHODS: The assay for measurement of TPMT activity in packed red blood cells is based on a non radioactive conversion of 6-thioguanine to 6-methylthioguanine. In 251 patients, TPMT activity was determined before and after a 4-h period of hemodialysis. In 49 patients (26 on azathioprine, 23 on mycophenolate mofetil as control group), TPMT activity was regularly determined during the first 120 days after renal transplantation. RESULTS: TPMT activity is elevated in red blood cells of uremic patients before hemodialysis when compared with TPMT activity after hemodialysis. The latter is comparable to the activity in healthy subjects. In patients treated with azathioprine, the TPMT activity showed a slow increase that declined to pre treatment values when azathioprine was withdrawn. This could not be observed in patients treated with mycophenolate mofetil. CONCLUSIONS: In uremic patients, TPMT activity is activated by some uremic factors that are removed by hemodialysis. In contrast to what has been observed before, dialysis shifted the TPMT activity close to that of a healthy control group. In patients treated with azathioprine after renal transplantation, the observed increase of TPMT activity could possibly be the result of enzyme induction. PMID- 11417445 TI - CYP2C19 polymorphism is not important for the in vivo metabolism of selegiline. AB - OBJECTIVES: To address the relevance of cytochrome P-450 (CYP) 2C19 polymorphism for the pharmacokinetics and dynamics of selegiline and its two known primary metabolites, desmethylselegiline and l-methamphetamine. METHODS: Six extensive (mephenytoin S/R ratio < 0.3; EM) and six poor (mephenytoin S/R ratio > 0.8; PM) hydroxylators of S-mephenytoin ingested a single 10-mg oral dose of selegiline hydrochloride. Serum concentrations of selegiline, desmethylselegiline and l methamphetamine were measured by gas chromatography--mass spectrometry for up to 48 h. In addition, the platelet monoamine oxidase type B (MAO-B) activity was measured for 14 days to describe possible differences in the pharmacodynamics of selegiline and its metabolites between EM and PM. RESULTS: The CYP2C19 phenotype had no significant effects on the pharmacokinetic variables of selegiline. PM of S-mephenytoin had 68% higher mean AUC of desmethylselegiline (P = 0.0017) than EM, but no significant differences were observed in other pharmacokinetic parameters of desmethylselegiline. Contrary to desmethylselegiline, the serum l methamphetamine concentrations were slightly lower in PM, but no statistically significant differences were observed in l-methamphetamine pharmacokinetics between the two CYP2C19 phenotypes. Accordingly, the magnitude of MAO-B inhibition showed no significant differences between the study groups. CONCLUSIONS: CYP2C19 polymorphism does not seem to be crucial for the metabolism or clinical effects of selegiline. PMID- 11417446 TI - An optimized methodology for combined phenotyping and genotyping on CYP2D6 and CYP2C19. AB - A method for simultaneous phenotyping and genotyping for CYP2D6 and CYP2C19 was tested. Six healthy volunteers were selected (three extensive and three poor metabolisers for CYP2D6). CYP2D6 was probed with dextromethorphan and metoprolol and CYP2C19 was probed with omeprazole. Blood samples were collected and analysed for dextromethorphan, dextrorphan, metoprolol, alpha-hydroxymetoprol, omeprazole and 5-hydroxyomeprazole by HPLC. Genotyping was performed for both CYP2D6 and CYP2C19. Generally, plasma levels could be measured up to 8 h post-dose except for alpha-hydroxymetoprolol in poor metabolizers (PMs) and dextromethorphan in extensive metabolizers (EMs) (35% below quantification limit). The correlation between the metabolic ratio based on timed individual measurements and the metabolic ratio based on the AUC0-12 values was significant at 3 h post-dose for all probes. In conclusion, the following procedure is suggested: administer metoprolol (100 mg) and omeprazole (40 mg); after 3 h, take a blood sample to assess the genotype and the metabolic ratio for CYP2D6 (metoprolol over alpha hydroxymetoprolol) and CYP2C19 (omeprazole over 5-hydroxyomeprazole) in plasma. With this procedure, all necessary information on the individual CYP2D6 and CYP2C19 metabolising capacity can be obtained in a practical, single-sample approach. PMID- 11417447 TI - Single- and multiple-dose pharmacokinetics of repaglinide in patients with type 2 diabetes and renal impairment. AB - OBJECTIVE: The primary objective of this single-centre, open-label, parallel group study was to evaluate the pharmacokinetics and safety profile of the prandial glucose regulator repaglinide, following single and multiple dosing, in patients with type 2 diabetes with and without varying degrees of renal impairment. METHODS: The study comprised three screening visits, followed by a 7 day inpatient period. Thirty-four patients, with normal renal function (n = 12), mild-to-moderate renal dysfunction (n = 12) or severe renal dysfunction (n = 10), received a single 2-mg dose of repaglinide on day 1, followed by preprandial 2-mg doses with main meals (breakfast, lunch and dinner) on each of days 2-4. A final 2-mg dose of repaglinide was administered on day 5. RESULTS: Patients with mild to-moderate renal impairment showed no significant differences in the pharmacokinetics of repaglinide, compared with patients with normal renal function. In the group of patients with severe renal dysfunction, the main pharmacokinetic finding was a longer half-life after multiple dosing. Rates of minor hypoglycaemia were similar in patients with severe, mild-to-moderate and no renal dysfunction. No major hypoglycaemic episodes occurred. CONCLUSION: Patients with type 2 diabetes and mild or moderate impairment of renal function may be treated with repaglinide without special precautions. If repaglinide is used in patients with severely impaired renal function, dose adjustment may be necessary if indicated by blood glucose measurements. PMID- 11417448 TI - Pharmacokinetic-pharmacodynamic model for the anticholinergic effect of glycopyrrolate. AB - OBJECTIVE: The purpose of this study was to develop and test a pharmacokinetic pharmacodynamic (PK-PD) model for the anticholinergic effect of glycopyrrolate in eight healthy male volunteers. METHODS: First, arterial drug concentration (Cp) data after a single intravenous (i.v.) bolus injection (5 micrograms/kg) were individually fitted to a three-compartment PK model. Second, the effect of a 2-h glycopyrrolate i.v. infusion (5 micrograms/kg/h) on the mean R-R interval (RRI) and the Hayano index of the high frequency variability of RRI (HF CCV) was modelled using an effect-compartment, inhibitory sigmoidal Emax model, with the individual PK parameters from the first part as constants. Third, the developed model was tested using a computer-driven infusion which aimed at two ascending steady-state effect-site concentrations (Ce) at 1-h intervals, corresponding to 20% and 80% of the maximal effect (Emax) observed in the second part. RESULTS: Modeling of the HF CCV data yielded the following mean (+/- SD) estimates: concentration at 50% of Emax (EC50), 2.46 +/- 0.58 ng/ml, equilibration half-time (t1/2 ke0), 42.5 +/- 7.7 min, and sigmoidicity factor (gamma), 7.26 +/- 2.82. The corresponding values for RRI data were 2.79 +/- 0.52 ng/ml, 58.3 +/- 17.2 min, and 4.75 +/- 1.56. During the computer-controlled two-step infusion (performed using HF CCV as the effect variable), the measured Cp approached the targeted Ce in most of the subjects, while the observed effect appeared to surpass the targeted levels. CONCLUSION: Although we were able to develop individual PK-PD models for glycopyrrolate, maintaining a stable anticholinergic effect in the computer-driven infusion appeared to be difficult. This is probably due to intra individual variability in the PK-PD parameters and the extremely steep concentration-effect relationship of glycopyrrolate. PMID- 11417449 TI - Drug prescribing among Danish children: a population-based study. AB - OBJECTIVES: To characterise the pattern of drug prescribing in Danish children below 19 years of age. METHODS: We used the Odense Pharmacoepidemiologic Database to identify all individual prescriptions for 0- to 19-year-olds in the County of Funen, Denmark, in 1998. The drug consumption was analysed by the defined daily dose methods and anatomical chemical classification system, using analytical templates for individual-based drug utilisation statistics. RESULTS: Of the examined population, 52.6% had one or more prescriptions issued. Overall, 10% of the children accounted for 67.5% of drug prescriptions. Approximately 40% of the children had a prescription issued before they were 1 year old. In 1- to 2-year olds, 85% received a prescription. Thereafter, the 1-year prevalence of drug use declined to 40%. The total drug consumption was similar between boys and girls until the early teens; thereafter the girls had a prescription issued twice as often as boys. In all age groups, respiratory drugs dominated and drugs used in dermatology was the second largest group. The third largest group was systemic anti-infective agents for children below 12 years of age, whereas it was psychotropic drugs among the oldest children. CONCLUSION: There is the same skewness of drug consumption among Danish children as among Danish adults, but the types of medication differ. Drug use is most extensive among children below 2 years but drug prescribing was considerable in all age groups. PMID- 11417450 TI - Is there an association between selective serotonin reuptake inhibitors and risk of abnormal bleeding? Results from a cohort study based on prescription event monitoring in England. AB - OBJECTIVE: To investigate whether an association between the use of selective serotonin reuptake inhibitor (SSRI) antidepressants and abnormal bleeding is demonstrated in a large population study. METHODS: An observational cohort study using cohorts from the Drug Safety Research Unit's prescription event monitoring database was performed. RESULTS: Analysis of combined haemorrhagic event rates calculated for the first 6 months of treatment for four SSRIs showed no significant difference between the rate for abnormal bleeding in the first month after starting treatment compared with months 2-6 [difference in rates 0.63 per 1000 patient months of treatment, 99% confidence interval (CI) -0.4, 1.67]. Comparison of the rates for the exposed combined SSRI cohort with the unexposed non-psychiatric drug cohort for the first month [relative risk (RR) 1.38, 95% CI 0.82, 2.34] and months 2-6 (RR 1.17, 95% CI 0.81, 1.68) showed no significant differences after adjustment for age and gender. However, there was a tendency towards highest risk with the combined SSRI cohort and lowest with the baseline cohort. CONCLUSION: This study provides weak evidence to support the hypothesis of a link between SSRIs and precipitation of bleeding events at a population level. The 95% CI is consistent with a possible risk of bleeding associated with SSRI users versus non-psychiatric drug users in the first month. Fuller consideration of confounding would be possible using follow-up of identified cases in a nested case-control study. PMID- 11417451 TI - The therapeutic course as a measure complementary to defined daily doses when studying exposure to antibacterial agents. AB - OBJECTIVE: The defined daily dose (DDD) method may not provide accurate information about exposure to antibacterial agents, owing to their short-term use and the consequently high turnover of subjects. This study was addressed to evaluate whether the number of "therapeutic courses" could offer additional information. METHODS: Prescription data of antibacterial agents (ATC J01) in Ravenna, Italy (350,000 inhabitants), were studied in 1998. All the prescriptions issued within 10 days were considered as belonging to one therapeutic course (TC). The following measures were compared: DDD/1000 inhabitants/year, number of exposed subjects/1000 inhabitants/year and number of TC/1000 inhabitants/year. In order to compare the number of 10-day TCs with the number of DDDs, the latter was divided by 10 (DDD10). The intensity of drug treatment was also evaluated as the average number of DDDs per TC (therapeutic intensity). RESULTS: Sixty-one percent of the subjects received only one prescription in 1998. Among the second prescriptions, 69% were issued after a time interval greater than 21 days. The overall prevalence of use was 329 per 1000 inhabitants, the TCs were 525 per 1000 inhabitants and DDD10/1000 inhabitants/year were 470. The therapeutic intensity varied with the age classes (being lowest in the elderly) and with the individual antibiotics used. Two or more antibacterials were used within the same TC in 7% of the cases. CONCLUSION: The measures considered in this study provided different estimates of exposure to antimicrobial agents. The combined analysis of DDDs and TCs may offer a more reliable information about exposure to antimicrobial agents. PMID- 11417452 TI - Termination of automatic atrial tachycardia in an infant by adequate sotalol dosing. Indication of clinically relevant age-dependent pharmacokinetics of sotalol. PMID- 11417453 TI - Florence Conference 2000. The joint CPT/EACPT conference in Florence, 16-20 July 2000. PMID- 11417454 TI - Effects of non-bronchoconstrictive doses of inhaled propranolol on airway responsiveness to methacholine. AB - OBJECTIVES: The aim of this study was to evaluate the effects of non bronchoconstrictive doses of propranolol on airway hyperresponsiveness to methacholine. METHODS: Double increasing concentrations (from 0.03 to 64 micrograms/ml) of inhaled propranolol were administered to a study population which included ten patients with mild asthma, ten rhinitics, and ten healthy control subjects. After the baseline bronchial responses to propranolol and methacholine, expressed as the cumulative provocative dose producing a 20% fall in forced expiratory volume in 1 s (PD20FEV1), were assessed, methacholine challenge was repeated after pretreatment with non-bronchoconstrictive doses of propranolol. RESULTS: The pharmacologically induced beta-blockade did not cause any effect in normal individuals, but it worsened airway responsiveness to methacholine in all asthmatics (geometric mean PD20 FEV1: 257 and 87 micrograms, respectively) and some rhinitics (geometric mean PD20 FEV1: 724 and 446 micrograms, respectively). CONCLUSION: Asthmatic patients were extremely sensitive to beta-blockers, whereas we observed a variable response to propranolol within the group of rhinitic subjects. This variability in the latter group is possibly because these individuals had different degrees of airway inflammation, increased parasympathetic activity, and beta-adrenoceptor dysfunction. PMID- 11417455 TI - Interlaminar differences in the pyramidal cell phenotype in cortical areas 7 m and STP (the superior temporal polysensory area) of the macaque monkey. AB - Pyramidal neurones were injected with Lucifer Yellow in slices cut tangential to the surface of area 7 m and the superior temporal polysensory area (STP) of the macaque monkey. Comparison of the basal dendritic arbors of supra- and infragranular pyramidal neurones (n = 139) that were injected in the same putative modules in the different cortical areas revealed variation in their structure. Moreover, there were relative differences in dendritic morphology of supra- and infragranular pyramidal neurones in the two cortical areas. Sholl analyses revealed that layer III pyramidal neurones in area STP had considerably higher peak complexity (maximum number of dendritic intersections per Sholl circle) than those in layer V, whereas peak complexities were similar for supra- and infragranular pyramidal neurones in area 7 m. In both cortical areas, the basal dendritic trees of layer III pyramidal neurones were characterized by a higher spine density than those in layer V. Calculations of the total number of dendritic spines in the "average" basal dendritic arbor revealed that layer V pyramidal neurones in area 7 m had twice as many spines as cells in layer III (4535 and 2294, respectively). A similar calculation for neurones in area STP revealed that layer III pyramidal neurones had approximately the same number of spines as cells in layer V (3585 and 3850 spines, respectively). Relative differences in the branching patterns of, and the number of spines in, the basal dendritic arbors of supra- and infragranular pyramidal neurones in the different cortical areas may allow for integration of different numbers of inputs, and different degrees of dendritic processing. These results support the thesis that intra-areal circuitry differs in different cortical areas. PMID- 11417456 TI - Mechanical properties of single motor units in the rabbit masseter muscle as a function of jaw position. AB - Positions and contractile properties of rabbit masseter motor units were investigated at different jaw gapes. Twitch responses were measured at gapes ranging from dental occlusion (0 degree) to maximum opening (21 degrees), in steps of 3 degrees. The twitches were elicited by stimulating motoneurons extracellularly in the trigeminal motor nucleus. The units appeared to produce a large variety of force vectors. On average motor units in the deep parts of the masseter produced considerably less twitch force (average: 25-30 mN) than those in the superficial parts (average: 45-50 mN) and anteriorly located motor units were slower than posteriorly located units. With an increase of jaw angle, twitches became slower, reflected by an increase (30%) of the twitch contraction time. Most motor units had a parabolic-like active jaw angle-force relationship. A large variation in the shape of the curves was found. The average optimum jaw angle was reached at 12 degrees jaw opening. In general, force output was relatively low (20-60% of maximum force) at occlusion and relatively high (60 100% of maximum force) at maximal jaw opening. Anteriorly and posteriorly located motor units differed significantly in their angle-force curves. Anteriorly located motor units produced less relative force at occlusion, showed a steeper increase of force with an increase of jaw angle, reached maximum force at larger jaw angles and produced larger forces at maximum jaw opening. The larger force changes in the more anterior units are probably related to their longer distance from the axis of jaw rotation. The large variability of motor unit properties and angle-force curves suggests that a fine gradation of both force magnitude and direction is possible within the masseter and that the angle-force curve of the whole muscle or of whole muscle parts is broader than that of individual motor units. This broadening may be considered as a mechanism to sustain active muscle force throughout a large movement range. PMID- 11417457 TI - Colinear facilitation promotes reliability of single-cell responses in cat striate cortex. AB - Behavior is controlled by neural activity in the brain. The final outcome of this neural control may critically depend on the firing reliability of individual neurons. A nearly constant, proportional relationship is usually found between the response mean and response variance. Here we asked whether lateral interactions within striate cortex that modulate response magnitude also proportionately modify the response variance of cortical neurons. In many cases, response variability depended on stimulus organization: discrete flankers colinearly placed well outside the neuron's receptive field increased response magnitude without a proportional increase in variance, thus improving the neuron's response reliability. Since colinear flanker facilitation is often seen near the neuron's firing threshold, increased response reliability for weak stimuli may contribute to enhancing perceptual saliency. PMID- 11417458 TI - Corticospinal control of locomotor pathways generating extensor activities in the cat. AB - Interneuronal convergence of corticospinal and segmental pathways involved with the generation of extensor activities during locomotion was investigated in decerebrate and partially spinalized cats. L-dihydroxyphenylalanine (L-DOPA) was slowly injected until long-latency, long-lasting discharges could be evoked by the stimulation of contralateral flexor reflex afferents (coFRA) and the group I autogenetic inhibition was reversed to polysynaptic excitation in extensor motoneurons. Under these conditions, we stimulated in alternation the contralateral pyramidal tract (PT), group I afferents from knee and ankle extensor muscles, and both stimuli together. We did the same for the stimulation of PT and of coFRA. Clear polysynaptic EPSPs could be evoked from all three sources in 32 extensor motoneurons. Convergence was inferred from spatial facilitation, which occurred when the amplitude of the EPSPs evoked by the combined stimuli was notably larger than the algebraic sum of the EPSPs evoked by individual stimulation. Spatial facilitation was found between PT and extensor group I inputs in 30/59 tests (51%) in 20 motoneurons and in all cases (6/6) between PT and coFRA in six motoneurons. When fictive locomotion was induced with further injection of L-DOPA, PT descending volleys from the same stimulating site could reset the stepping rhythm by initiating bursts of activity in all extensors. These results indicate that at least some of the corticospinal fibers project onto interneurons shared by the coFRA and the polysynaptic excitatory group I pathways to extensors. The implications of such convergence patterns on the organization of the extensor "half-center" for locomotion are discussed. PMID- 11417459 TI - Perception of angular head position during attempted alignment with eccentric visual objects. AB - The perception of angular head position with respect to a visual object was investigated using three different methods: Pointer indication (P); in the dark, subjects' (Ss') heads were horizontally turned to various positions (range +/- 54 degrees); Ss then rotated a pointer carrying a light emitting diode (LED) so as to align it with head position. Active head pointing (A); again in darkness, the pointer was rotated to various positions; Ss then turned their heads so as to align them with the pointer. Reading from visible scale (V); Ss viewed a degrees scale on a circular screen; Ss' heads were turned as in P, and Ss reported head position in terms of this scale. The results obtained with all three methods indicate that head position is overestimated with respect to the visual object (LED, scale mark): object position exceeded head position by 6, 18, and 7% when measured with the P-, A-, and V-methods, respectively (median values). The observed misalignment between head and object suggests that subjective primary eye position is shifted in the direction of head rotation by a cross-talk of head position. Whether a functional advantage or merely a tolerated side-effect of other constraints, this behavior confers the impression of looking "straight ahead" in the literal sense when gaze is shifted by coordinated eye-head movements with a 10% eye and a 90% head share in total lateral displacement. In an attempt to probe a hypothesized internal representation of head position implied in head-to-object alignment, Ss were also to estimate head position in space using only neck proprioceptive information. In complete darkness, responses were often non-linear functions of head position with overestimation of large eccentricities. When a head-centered LED was added (which conveyed no position information), responses became more linear, suggesting that the mere presence of visual afferents may improve the perceptual interpretation of proprioceptive information. PMID- 11417460 TI - Human development of grip force modulation relating to cyclic movement-induced inertial loads. AB - The present study examines the development of grip force modulation relating to self-induced loads during repetitive vertical arm movements at different frequencies with a hand-held object. One hundred and thirty-four 3- to 6-year-old children and 16 adults were asked to move a lightweight object up and down at increasing rates from 0.5 Hz up to individual maximal arm movement rates (> 2.5 Hz). Grip forces were measured by a uni-axial force transducer and the inertial forces (tangential forces) were calculated from the measurements by accelerometers within the object. Generally, the quality of anticipatory grip force control improved from movement frequencies of 1 Hz to 2.5 Hz and decreased above 2.5 Hz. At movement frequencies below 1 Hz, the phase lag between grip and load force cycles was longer in children than in adults (children: median = 17 ms; adults: median = 0 ms); however, there were no significant differences between the age groups above 1.5 Hz. The grip-to-load force ratio and the grip force modulation were not significantly different from the adults up to 2.5 Hz. The fine coupling of grip and load forces (precise temporal and gain control) showed age effects among the 3- to 6-year-old children, mainly between 3- and 4 year-olds, and in adults extremely good coupling at medium arm movement frequencies. However, concerning the pure temporal coordination of grip and inertial forces, there were only differences between the 3-year-old children and the other groups at arm movement frequencies above 2.5 Hz. The results show that, during cyclic movements with hand-held loads, temporal control is well established at the age of 4 years whereas the fine gain control needs a longer time to develop. The anticipatory control is dependent on the arm movement frequency and, therefore, how rapidly the inertial loads change. The high level of anticipatory control during self-induced repetitive actions in children of 3-6 years of age is interpreted as an early developing ability to predict precisely cyclic self-induced inertial loads of hand-held objects when the object properties are known by proprioception. The timing, which is possibly related to cerebellar functions, is, to some extent, developed earlier than the gain control, which may be associated with cortical functions. PMID- 11417461 TI - Amygdala, hippocampus, and unconditioned fear. AB - Embedded within contemporary views of emotional learning is a well-founded agreement that the amygdala plays a pivotal role in the formation and consolidation of aversive memories formed during fear conditioning. However, it is important to determine whether observed deficits are reflective of a memory impairment or whether they are simply attributable to a deficit in the performance of unconditioned fear responses such as freezing. Within the neurobiology of learning and memory literature, there is an ongoing debate concerning the potential role of the amygdala in the performance of unconditioned fear responses. A view put forth by Vazdarjanova and McGaugh (1998) suggests that the amygdala is not required for the formation and consolidation of the aversive memories formed during fear conditioning, but is essential in the performance of unconditioned fear responses. Data provided by Maren (1999) counter this view by positing that the amygdala is not required for the performance of fear responses, but its role is of a mnemonic nature in the conditioning of fear to neutral cues. To clarify the amygdala's participation in these two processes, a useful approach would involve a situation where animals with amygdala damage were examined for their unconditioned fear responses in reaction to footshock as well as the conditioning of these reactions to previously neutral cues paired with the aversive event. We have previously reported that rats with amygdala or hippocampal damage are impaired in discriminative fear conditioning to context. In the present experiment, we report the initial unconditioned fear responses to footshock by these same animals as well as the conditioned responses during testing. In both groups, the fear responses assessed (freezing, urination, defecation, and locomotion) were not impaired and did not differ from those expressed by the sham animals. The impairment of discriminative fear conditioning to context, in combination with the present experiment, represents a dissociation where damage to specific memory structures (amygdala or hippocampus) debilitates the mnemonic processes involved in fear conditioning, but not the performance of the fear responses per se. PMID- 11417462 TI - The influence of postural threat on the control of upright stance. AB - In order to utilize static posturography as a tool with which to diagnose pathological balance disorders, it is necessary to understand the contribution of psychological factors such as fear of falling. In this study we examined kinetic and kinematic parameters during quiet stance of eight young healthy adults standing under three conditions which posed increasing levels of postural threat. Participants were required to stand quietly, both with and without visual information and with normal or reduced vestibular information for 2-min periods at each of three surface conditions: ground level (low threat), 81 cm above the ground completely surrounded by a support surface (medium threat) and 81 cm above ground at the edge of the surface (high threat). In addition to calculating displacement and frequency measures for movements of centre of pressure (COP) and centre of mass (COM), a measure of stiffness was derived from the difference between COP-COM signal. Mean EMG activity was recorded bilaterally from anterior and posterior muscles of the lower and upper leg. A stiffening strategy was adopted when individuals stood under high threat conditions involving significant changes in kinematic, kinetic and EMG variables. The A-P stiffness constant increased significantly (27.5%) for the high threat compared to low threat condition, independent of vision or vestibular information. Changes in stiffness were accompanied by: (1) backward shift of the mean A-P position of COP and COM, (2) increased mean power frequency and decreased amplitude displacement of the COP and (3) decreased amplitude displacements of the COM. Of the significant changes observed in mean EMG activity with increased postural threat, only tibialis anterior was significantly correlated with increased stiffness, while activity in soleus, medial gastrocnemius and rectus femoris was highly correlated with mean position of COP. PMID- 11417463 TI - Multijoint grasping movements. Simulated and observed effects of object location, object size, and initial aperture. AB - Studies of human prehension have revealed characteristic patterns of grasping kinematics. We sought to gain insight into the determinants of those patterns by means of a computer simulation and accompanying behavioral experiment concerning multijoint, planar grasping behavior. The simulation was based on a recent theory of posture-based motion planning which hypothesizes that movement preparation entails time-limited, multiple task-constraint satisfaction. Prehension was modeled with a stick-figure animation involving 12 series of 81 grasping movements. Circular objects to be grasped were located at three angles (45 degrees, 90 degrees, and 135 degrees) and at three distances (20 cm, 30 cm, and 40 cm) relative to the initial location of the hand in the workplane. Additionally, three object sizes (2 cm, 4 cm, and 6 cm in diameter) and three initial aperture sizes (0.3 cm, 3.3 cm, and 7.0 cm) were used. Analyses of the simulated grasping movements focused on the time course of the hand opening, the tangential velocity of the wrist, and the rotations of the joints in the arm, hand, and fingers. The results showed that the model accurately mimicked detailed kinematics of prehension observed in earlier studies. With respect to the frequently reported relationship between object size and hand opening, the simulations further revealed an effect of initial aperture. This predicted effect was confirmed in an experiment in which four participants performed analogous planar grasping tasks. An analysis of the time course of the opening of the hand showed that maximum aperture covaried with initial aperture. A conclusion of this work is that a major determinant of grasping kinematics is avoidance of collisions with objects that are to be grasped. PMID- 11417464 TI - Cortical representation of whole-body movement is modulated by proprioceptive discharge in humans. AB - Previous studies have revealed the influence of ongoing sensory discharge on modulating the central representation of muscle afferents from individual limbs. In the present study, we explored the potential for such modulatory influence on the afferent discharge arising from induced whole-body movement. Vestibular and somato-sensory inputs arise from such whole-body movement. The convergence of these two modalities is important in motor control, especially for the maintenance of postural stability. We hypothesised that transmission of proprioceptive and vestibular information to the cortex would be reduced as a result of muscle-spindle discharge in knee extensor muscles. Perturbation-evoked responses (PERs), recorded from central scalp electrodes (C3, CZ, C4), were evoked through rapid translations of subjects who were seated in a chair on a movable platform. PERs were recorded during passive linear translations alone and preceded by vibration of the patellar tendon. The PER was characterised by a slow, negative potential peaking at approximately 150 ms (N150) following displacement of the chair. The amplitude of the PER was reduced following vibration to 56% of the control. Such reduction of PERs was comparable to the attenuation of somatosensory evoked potentials and soleus H-reflex magnitudes from tibial-nerve stimulation. We conclude that muscle-spindle discharge in knee extensor muscles leads to gating of both of these afferent pathways. These results have potential implications to the understanding of the CNS control of stability during ongoing movement. PMID- 11417465 TI - Perturbed upper limb movements cause short-latency postural responses in trunk muscles. AB - Addition of a load to a moving upper limb produces a perturbation of the trunk due to transmission of mechanical forces. This experiment investigated the postural response of the trunk muscles in relation to unexpected limb loading. Subjects performed rapid, bilateral shoulder flexion in response to a stimulus. In one third of trials, an unexpected load was added bilaterally to the upper limbs in the first third of the movement. Trunk muscle electromyography, intra abdominal pressure and upper limb and trunk motion were measured. A short-latency response of the erector spinae and transversus abdominis muscles occurred approximately 50 ms after the onset of the limb perturbation that resulted from addition of the load early in the movement and was coincident with the onset of the observed perturbation at the trunk. The results provide evidence of initiation of a complex postural response of the trunk muscles that is consistent with mediation by afferent input from a site distant to the lumbar spine, which may include afferents of the upper limb. PMID- 11417466 TI - Predicting sensory events. The role of the cerebellum in motor learning. AB - There is growing evidence that the cerebellum is involved in the implicit learning of movement sequences. On the serial reaction time (RT) task patients with cerebellar lesions fail to demonstrate normal decreases in RT and we have shown a similar effect in monkeys with bilateral cerebellar lesions. However, it is not clear if this impairment is unique to sequence learning or whether the cerebellum is also involved in the learning of discrete responses to predictable visual targets. We investigated this possibility in another group of monkeys with bilateral lesions of the cerebellum centred on the lateral nuclei. Three animals were pre-operatively trained to make rapid manual responses to a single target appearing on a touch-sensitive VDU screen. In one condition, a target could appear at any of three possible locations (spatially unpredictable). In a second condition the target always appeared in the same place (spatially predictable). A third condition was similar to the second except that the onset of the target was temporally predictable whereas in the previous conditions this parameter was randomized. Following the lesions, the RT savings earned on the conditions in which the cues were predictable were abolished. This was despite a lack of significant increase in movement times. The results imply that the animals were either failing to predict the spatial location or time of presentation of the target, or that they were unable to use their prediction to improve their reaction times. The function of the cerebellum in motor sequence learning may therefore be part of a more general operation in learning to prepare responses to predictable sensory events. PMID- 11417467 TI - Changes of error-related ERPs with age. AB - Errors in reaction tasks are followed by a negative component of the event related brain potential (ERP), the error negativity (Ne), which is thought to be a correlate of error detection. In the present study we show that, in tasks that induce different types of errors, the amplitude of the Ne was reduced in elderly (54-65 years old) compared with young subjects (19-25 years old). This reduction was also seen in single trials, as were computed for one of the visual tasks. Moreover, in this data set, the single-trial Ne was also delayed for the elderly compared with the young. These data suggest an alteration of error detection in the elderly, which is only marginally reflected in performance. PMID- 11417468 TI - Cerebellar lesions impair context-dependent adaptation of reaching movements in primates. AB - To produce accurate movements when conditions change suddenly, the brain must be capable of learning multiple versions of a given motor task and must be able to access the appropriate program using sensory information linked to the context of the movement. The neural basis for context-dependent motor learning is uncertain, but the cerebellum is thought to play a fundamental role. In this study, we examined the effect of lesions of the dorsal vermal and paravermal cerebellar cortex on the adaptation of reaching movements produced by modified visual feedback and accessed with a visual cue. Two rhesus monkeys were trained to point to targets displayed on a video monitor while viewing monocularly with either eye. During the experimental sessions, visual information received by one eye (the "modified" eye) was displaced horizontally, while the information received by the other ("normal") eye remained unaltered. In the first set of experiments (noncontextual paradigm), the animals pointed to targets while viewing with the modified eye. This paradigm resulted in a gradual improvement in pointing accuracy when viewing with that eye, but also produced a shift in pointing responses of equivalent size when viewing with the normal eye. In the second set of experiments (contextual paradigm), the animals alternated six blocks of reaches while viewing monocularly with the modified eye with six blocks viewing with the normal eye. This paradigm improved the pointing accuracy when viewing with the modified eye, but produced only a small shift in pointing responses when viewing with the normal eye. After the dorsal vermal and paravermal cerebellar cortex were resected, no change occurred in the pattern of adaptation produced by the noncontextual paradigm. The contextual paradigm, however, no longer selectively adapted pointing responses for each eye, but rather produced a pointing shift of equivalent size when viewing with either eye. The results indicate that pointing responses can be differentially adapted for each viewing eye, which is a form of context-dependent motor learning. This capability was lost after focal lesions of the dorsal vermal and paravermal cerebellar cortex, suggesting that these regions of cerebellar cortex are required to learn or store multiple representations of a movement, or to retrieve the appropriate motor program in a given sensory context. PMID- 11417469 TI - The effect on corticospinal volleys of reversing the direction of current induced in the motor cortex by transcranial magnetic stimulation. AB - Descending corticospinal volleys were recorded from a bipolar electrode inserted into the cervical epidural space of four conscious human subjects after monophasic transcranial magnetic stimulation over the motor cortex with a figure of-eight coil. We examined the effect of reversing the direction of the induced current in the brain from the usual posterior-anterior (PA) direction to an anterior-posterior (AP) direction. The volleys were compared with D waves evoked by anodal electrical stimulation (two subjects) or medio-lateral magnetic stimulation (two subjects). As reported previously, PA stimulation preferentially recruited I1 waves, with later I waves appearing at higher stimulus intensities. AP stimulation tended to recruit later I waves (I3 waves) in one of the subjects, but, in the other three, I1 or D waves were seen. Unexpectedly, the descending volleys evoked by AP stimulation often had slightly different peak latencies and/or longer duration than those seen after PA stimulation. In addition the relationship between the size of the descending volleys and the subsequent EMG response was often different for AP and PA stimulation. These findings suggest that AP stimulation does not simply activate a subset of the sites activated by PA stimulation. Some sites or neurones that are relatively inaccessible to PA stimulation may be the low-threshold targets of AP stimulation. PMID- 11417470 TI - Gravitational cues contribute to accurate localisation of mentally represented cutaneous targets. AB - The purpose of this research was to ascertain whether gravitational cues contribute to localisation ability of mentally represented cutaneous targets. Eight right-handed male subjects were asked to use each hand to point accurately toward four points in the anterior trunk midline solely on the basis of the mental representation of their position along the trunk while standing upright and while lying supine. It was found that subjects were more accurate when performing in the upright posture. However, statistical analysis of the mean constant transverse errors of pointing showed that neither posture nor target factors had any significant effect on pointing accuracy. On the contrary, analysis of the mean constant longitudinal errors showed that both the posture and target factors were significant. Subjects pointed caudally to the target locations and made the largest errors, which were on the order of centimeters, when performing in the horizontal posture. These findings indicate that gravitational cues are critical in accurately localising mentally represented cutaneous targets. PMID- 11417471 TI - Unwinding the loop of Bcl-2 phosphorylation. AB - Recent evidence indicates that anti-apoptotic functions of BcI-2 can be regulated by its phosphorylation. According to the 'mitotic arrest-induced' model, multi site phosphorylation of the BcI-2 loop domain is followed by cell death. In contrast, in cytokine-dependent cell lines, cytokines mediate phosphorylation of BcI-2 on S70, preventing apoptosis. As discussed in this review, these models are not mutually exclusive but reflect different cellular contexts. During mitotic arrest, signal transduction is unique and is fundamentally different from classical mitogenic signaling, since the nucleus membrane is dissolved, gene expression is reduced, and numerous kinases and regulatory proteins are hyperphosphorylated. Hyperphosphorylation of BcI-2 mediated by paclitaxel and other microtubule-active drugs is strictly dependent on targeting microtubules that in turn cause mitotic arrest. In addition to serine-70 (S70), microtubule active agents promote phosphorylation of S87 and threonine-69 (T69), inactivating BcI-2. A major obstacle for identification of the mitotic BcI-2 kinase(s) is that inhibition of putative kinase(s) by any means (dominant-negative mutants, antisense oligonucleotides, pharmacological agents) may arrest cycle, preventing mitosis and BcI-2 phosphorylation. The role of BcI-2 phosphorylation in cell death is discussed. PMID- 11417472 TI - Nucleoside analogues: mechanisms of drug resistance and reversal strategies. AB - Nucleoside analogues (NA) are essential components of AML induction therapy (cytosine arabinoside), effective treatments of lymphoproliferative disorders (fludarabine, cladribine) and are also used in the treatment of some solid tumors (gemcitabine). These important compounds share some general common characteristics, namely in terms of requiring transport by specific membrane transporters, metabolism and interaction with intracellular targets. However, these compounds differ in regard to the types of transporters that most efficiently transport a given compound, and their preferential interaction with certain targets which may explain why some compounds are more effective against rapidly proliferating tumors and others on neoplasia with a more protracted evolution. In this review, we analyze the available data concerning mechanisms of action of and resistance to NA, with particular emphasis on recent advances in the characterization of nucleoside transporters and on the potential role of activating or inactivating enzymes in the induction of clinical resistance to these compounds. We performed an extensive search of published in vitro and clinical data in which the levels of expression of nucleoside-activating or inactivating enzymes have been correlated with tumor response or patient outcome. Strategies aiming to increase the intracellular concentrations of active compounds are presented. PMID- 11417473 TI - Magnetic resonance imaging detection of avascular necrosis of the bone in children receiving intensive prednisone therapy for acute lymphoblastic leukemia or non-Hodgkin lymphoma. AB - The purpose of this study was to determine the frequency with which magnetic resonance (MR) imaging detects avascular necrosis of the bone (AVNB) in children with acute lymphoblastic leukemia (ALL) or advanced-stage non-Hodgkin lymphoma (NHL) who receive prednisone during remission induction, reinduction, and maintenance chemotherapy; to assess the clinical significance of these findings; and to identify factors predictive of AVNB. We prospectively obtained MR imaging of the hips and knees of 116 children who had completed at least 1 year of treatment for ALL or advanced-stage NHL on identical prednisone-containing regimens between December 1991 and October 1994. MR imaging findings of AVNB were compared with clinical outcomes, and the effect of therapeutic and patient factors on the frequency of AVNB was analyzed. The MR imaging findings of 17 of the 116 participating patients were consistent with AVNB. The most common clinical manifestation was joint pain (11 patients). Only one patient had progressive joint deterioration that necessitated surgical replacement. Only age 10 years or more at the time of the primary diagnosis was significantly associated with the development of AVNB (P = 0.004). MR imaging showed changes consistent with AVNB in approximately 15% of this patient population. However, most patients in this study who had MR imaging signs of AVNB did not experience progressive joint destruction, even with continued prednisone therapy. Therefore, the clinical usefulness of MR imaging as a screening tool for AVNB in this set of patients remains uncertain. PMID- 11417474 TI - Daunorubicin continuous infusion induces more toxicity than bolus infusion in acute lymphoblastic leukemia induction regimen: a randomized study. AB - We report the first randomized study assessing the efficacy and safety of daunorubicin (DNR) continuous infusion (CI) compared to the more conventional 30 min infusion (i.v.) in newly diagnosed adult acute lymphoblastic leukemia (ALL). Seventy-seven patients were initially randomized to receive either a 24-h CI DNR (60 mg/m2 days 2-4) (40 patients) or bolus DNR at the same dosage (37 patients) with vincristine (2 mg i.v. days 1, 8, 15) and oral prednisone (60 mg/m2 days 1 15), without hematopoietic growth factor support, as an induction regimen. The distribution of adverse prognostic factors was comparable in the two-induction arm. Acute toxicity was more important in the CI arm. Gram negative infection (9 vs 1 gram negative septicemia, P = 0.01) and infection-related deaths (6 vs 1 deaths, P = NS) occurred more frequently in the CI arm during the induction treatment than in the i.v. arm, leading to the study interruption. Neutropenia but not thrombopenia duration was significantly longer in the CI arm than in the i.v. arm (18 days vs 14 days, P > 0.05 and 16 days vs 12 days, P > 0.05, respectively). Despite a similar CR rate according to the method of DNR administration (68% in the CI DNR arm vs 76% in the i.v. arm after the first course), there was a trend toward higher freedom from relapse (FFR) after DNR CI (48% vs 28% in the i.v. arm at 5 years, P = NS), suggesting that despite this high toxicity, DNR CI may improve the CR quality and decrease further the residual disease. PMID- 11417475 TI - The prognostic value of cytogenetics is reinforced by the kind of induction/consolidation therapy in influencing the outcome of acute myeloid leukemia--analysis of 848 patients. AB - We studied the impact of cytogenetics and kind of induction/consolidation therapy on 848 adult acute myeloid leukemia (AML) patients (age 15-83). The patients received three types of induction/consolidation regimen: standard (daunorubicin and cytosine arabinoside (3/7); two cycles); intensive (idarubicin, cytosine arabinoside and etoposide (ICE), plus mitoxantrone and intermediate-dose Ara-C (NOVIA)); and low-dose (low-dose cytosine arabinoside). CR patients under 60 years of age, if an HLA-identical donor was available received allogeneic stem cell transplantation (allo-SCT); otherwise, as part of the program, they underwent autologous (auto)-SCT. CR rates significantly associated with 'favorable' (inv(16), t(8;21)), 'intermediate' ('no abnormality', abn(11q23), +8, del(7q)) and 'unfavorable' (del (5q), -7, abn(3)(q21q26), t(6;9), 'complex' (more than three unrelated cytogenetic abnormalities)) karyotypes (88% vs 65% vs 36%, respectively; P = 0.0001). These trends were confirmed in all age groups. On therapeutic grounds, intensive induction did not determine significant increases of CR rates in any of the considered groups, with respect to standard induction. Low-dose induction was associated with significantly lower CR rates. Considering disease-free survival (DFS), multivariate analysis of the factors examined (including karyotype grouping) showed that only age > 60 years significantly affected outcome. However, in cases where intensive induction was adopted, 'favorable' karyotype was significantly related to longer DFS (P = 0.04). This was mainly due to the favorable outcome of t(8;21) patients treated with intensive induction. Patients receiving allo-SCT had significantly longer DFS (P = 0.005); in particular, allo-SCT significantly improved DFS in the 'favorable' and 'intermediate' groups (P = 0.04 and P = 0.048, respectively). In conclusion our study could provide some guidelines for AML therapy: (1) patients in the 'favorable' karyotype group seem to have a longer DFS when treated with an intensive induction/consolidation regimen, adopted before auto-SCT instead of standard induction; this underlines the importance of reinforcement of chemotherapy, not necessarily based on repeated high-dose AraC cycles. Allo-SCT, independently of induction/consolidation therapy, should be considered an alternative treatment; (2) patients in the 'intermediate' karyotype group should receive allo-SCT; (3) patients in the 'unfavorable' karyotype group should be treated using investigational chemotherapy, considering that even allo-SCT cannot provide a significantly longer DFS, but only a trend to a better prognosis. PMID- 11417476 TI - Identification and molecular characterisation of a CALM-AF10 fusion in acute megakaryoblastic leukaemia. AB - The t(10;11)(p13;q14-21) is a non-random translocation described in acute lymphoblastic and myeloid leukaemias. It results in the fusion of the gene CALM, which encodes a clathrin assembly protein, on 11q14 to the gene AF10, a putative transcription factor on 10p13. Here we describe for the first time, the occurrence of a CALM-AF10 fusion in a case of acute megakaryoblastic leukaemia. Fluorescence in situ hybridisation and reverse transcriptase polymerase chain reaction were used to confirm the presence of a CALM-AF10 fusion. A novel splice variant of CALM missing nt 1927-2091 was also detected. Though CALM is a cytoplasmic protein, the chimaeric fusion product is able to localise to both the nucleus and cytoplasm. Analysis of the fusion variants suggests, however, that the critical fusion product is likely to be cytoplasmic and contain the interactive leucine zipper of AF10. PMID- 11417477 TI - t(7;12)(q36;p13) and t(7;12)(q32;p13)--translocations involving ETV6 in children 18 months of age or younger with myeloid disorders. AB - Our retrospective karyotype review revealed two rare recurrent translocations affecting ETV6 (TEL): t(7;12)(q36;p13) and t(7;12)(q32;p13). Five patients with a t(7;12) were from a group of 125 successfully karyotyped pediatric patients enrolled in consecutive clinical AML trials of the Dutch Childhood Leukemia Study Group over a period of 7 years. During a search of available cytogenetic databases, we found 7q and 12p abnormalities in two additional Dutch patients and in three participants in Pediatric Oncology Group trials. A del(12p) had been initially identified in four of these patients and re-examination of the original karyograms revealed a t(7;12)(q36;p13) in two instances and a probable t(7;12) in the other two. FISH confirmed the presence of a t(7;12)(q36;p13) in the latter. Most (n = 7) also had trisomy 19. The t(7;12)(q36;p13) (n = 9) was more common than the t(7;12)(q32;p13) (n = 1). These subtle translocations were found only in children 18 months of age or younger. A literature search revealed that the t(7;12) with break-points at 7q31-q36 and 12p12-p13 had been reported in six children with myeloid disorders and in two with acute lymphoblastic leukemia; all were 12 months of age or younger. Only two of the 17 for whom survival data were available, were alive after at least 22 months of continuous complete remission. Our findings suggest that ETV6 rearrangements due to a t(7;12) may play an adverse role in myeloid disorders in children 18 months of age or younger. Therefore, children in this age group with myeloid disorders should be screened for both MLL and ETV6 rearrangements. PMID- 11417478 TI - In vitro susceptibility to TRAIL-induced apoptosis of acute leukemia cells in the context of TRAIL receptor gene expression and constitutive NF-kappa B activity. AB - The TNF-related apoptosis-inducing ligand (TRAIL) is currently under evaluation as a possible (co-)therapeutic in cancer treatment. We therefore examined 129 cell samples from patients with de novo acute leukemia as to their constitutive susceptibility to TRAIL-induced apoptosis In vitro. Only 21 (16%) cell samples revealed at least 10% TRAIL-susceptible cells/sample as detected by flow cytometric annexinV staining after 24 h culture compared with medium control. Precursor B cell ALL samples (11 (27%) of 41) were more TRAIL-susceptible compared with AML (5 (9%) of 54; P < 0.05) but not compared with precursor T cell ALL (5 (15%) of 34; P = 0.20). Furthermore, we examined constitutive mRNA expression levels of TRAIL receptors R1-R4 by semi-quantitative RT-PCR (n = 58). Expression levels were heterogeneous, however, there was no significant correlation between the expression of the signal-transducing receptors (R1, R2) as well as of the decoy receptors (R3, R4) and TRAIL sensitivity in this series. Constitutive NF-kappa B activity has been shown to influence TRAIL susceptibility of leukemic cells. In 39 leukemic cell samples examined, we found a generally high NF-kappa B activity as detected by electrophoretic mobility shift assay which did not differ between TRAIL-susceptible and TRAIL-resistant cases. Finally, 49 acute leukemic cell samples were coincubated with doxorubicin in vitro. Doxorubicin sensitized four of 35 initially TRAIL-resistant samples and augmented TRAIL-induced apoptosis in two of 14 TRAIL-susceptible samples. In summary, constitutive TRAIL susceptibility differs between leukemia subtypes and does not correlate with mRNA expression levels of the TRAIL receptors R1-R4 as well as constitutive NF-kappa B activation status. The observed sensitization of leukemic cells to TRAIL by doxorubicin in vitro indicates that TRAIL should be further evaluated as to its possible role as an in vivo cotherapeutic in acute leukemia. PMID- 11417479 TI - High level resistance to glucocorticoids, associated with a dysfunctional glucocorticoid receptor, in childhood acute lymphoblastic leukemia cells selected for methotrexate resistance. AB - The molecular basis for the clinical presentation of broad-range drug resistance in childhood ALL is poorly understood. In this study, high level cross-resistance to the glucocorticoid dexamethasone was encountered in a childhood ALL cell line selected for resistance to methotrexate (CEM MTX-R3). Compared with wild-type (WT) CEM cells, MTX-R3 cells had significantly fewer glucocorticoid binding sites, as well as reduced glucocorticoid receptor protein and mRNA levels. DNA sequencing and restriction fragment-length polymorphism (RFLP) analysis showed that WT cells expressed both a wild-type and a mutant (GR753F) glucocorticoid receptor allele, while MTX-R3 cells expressed only the GR753F allele. Therefore, the cross-resistance of MTX-R3 cells to dexamethasone appeared due to loss of expression of the wild-type glucocorticoid receptor allele. In an effort to gain insight into the underlying basis for the development of cross-resistance to methotrexate and glucocorticoids, glucocorticoid receptor nuclear translocation experiments were carried out. Exposure of WT cells to either dexamethasone or the cytotoxic agents cytarabine and methotrexate caused translocation of the glucocorticoid receptor from the cytoplasm into the nucleus. These data indicate that exposure of childhood ALL cells to cytotoxic agents may result in ligand independent glucocorticoid receptor activation which, in the context of the outgrowth of drug-resistant cells, could lead to the co-selection of glucocorticoid resistance. PMID- 11417480 TI - Treatment with inhibitors of caspases, that are substrates of drug transporters, selectively permits chemotherapy-induced apoptosis in multidrug-resistant cells but protects normal cells. AB - Many chemotherapeutic agents induce apoptosis in tumor cells, but killing of normal cells remains a major obstacle. Development of multidrug resistance further limits chemotherapy in cancer. Here, I show that multidrug resistance can be exploited for selective killing of multidrug-resistant cells by a combination of an apoptosis-inducing agent that is not a substrate of either Pgp or MRP (e.g. flavopiridol) with a caspase inhibitor that is a substrate (e.g. Z-DEVD-fmk). In normal cells, treatment with caspase inhibitors prevented PARP cleavage, nuclear fragmentation, and cell death caused by flavopiridol or epothilone B. In contrast, Pgp- and MRP-expressing cells were not rescued by caspase inhibitors. Furthermore, reversal of drug resistance renders Pgp cells sensitive to caspase inhibitors abolishing therapeutic advantage. Thus, caspase inhibitors, that are inactive in multidrug-resistant cells, protect normal but not multidrug-resistant cells against chemotherapy, permitting selective eradication of multidrug resistant cells. Clinical application of this approach may diminish the toxic side-effects of chemotherapy in patients with multidrug-resistant tumors. PMID- 11417481 TI - TGF beta-induced SMAD2 phosphorylation predicts inhibition of thymidine incorporation in CD34+ cells from healthy donors, but not from patients with AML after MDS. AB - Cells from patients with MDS-derived AML display heterogeneous proliferative responses to transforming growth factor beta (TGF beta). We analyzed growth inhibition and SMAD2 phosphorylation by TGF beta in CD34+ cells from nine patients, as compared to normal controls. While TGF beta consistently inhibited thymidine incorporation of normal cells (41% of control, P < 0.05), cells from patients with AML were growth-inhibited in only four of seven cases (40%), whereas TGF beta stimulated thymidine incorporation in the three other samples (166%). Remarkably, TPO reverted the stimulatory effect of TGF beta to profound growth inhibition. Upon exposure to TGF beta, SMAD2 protein was phosphorylated in normal CD34+ cells (n = 3), CD34+ leukemic blasts from all examined patients with AML (n = 4), and in the myeloid leukemic cell lines M-07e and HEL. TGF beta inhibited TPO-mediated thymidine incorporation, cell proliferation and survival in all samples analyzed. In M-07e cells and CD34+ cells from healthy donors, this inhibition was enhanced by an antagonist of JAK2 (AG490), but not a MEK-1 antagonist (PD098059). Conversely, in CD34+ cells from a patient with AML, both AG490 and PD098059 significantly enhanced TGF beta-mediated suppression of TPO induced thymidine incorporation. Thus, in MDS-derived AML, altered responses to TGF beta may be due to defects downstream of SMAD2 and may involve MAPK activation. PMID- 11417482 TI - Role of splenectomy in the treatment of myelodysplastic syndromes with peripheral thrombocytopenia: a report on six cases. AB - Thrombocytopenia is generally of central origin in MDS, but can be due to peripheral platelet destruction in some cases. We studied platelet lifespan in 61 MDS cases with platelets < 70,000/mm3 and marrow blasts < 10%. Nine of them (15%) had a major platelet lifespan reduction (< 3.5 days), and were considered for splenectomy. Three of them were not splenectomized due to rapid death, patient refusal and older age plus liver predominance of platelet sequestration, respectively. The remaining six patients (two females and four males, median age 50 years, range 32 to 65) were splenectomized 3 to 21 months after diagnosis. Before splenectomy, five of them had RA and one had CMML. Platelets counts ranged from 5000 to 30,000/mm3 and did not durably respond to other treatments. Three of the patients has a relapse of platelet counts, concomitantly required platelet transfusion due to recurrent blending, whereas three had anemia (two required erythrocyte transfusion) and four had neutropenia. Three months after surgery, platelet counts ranged from 55,000 to 160,000/mm3 (> 100,000/mm3 in four cases), no patient required platelet or erythrocyte transfusion, but there was no effect on neutrophil counts. Three patients had a relapse of platelet counts, concomitant with progression to AML in two of them, whereas the third relapsing case achieved normal platelet counts with further danazol. One patient died with normal platelet counts 12 months after splenectomy (from sepsis, probably related to neutropenia rather than splenectomy). Two patients remained with normal platelet counts 10 and 52 months after surgery. Our findings suggest that the mechanism of thrombocytopenia should be studied more often in 'low risk' MDS (i.e. with low bone marrow blast counts) with thrombocytopenia, as about 15% of them appear to have peripheral platelet destruction. Some of those patients may benefit from splenectomy. PMID- 11417483 TI - Deletions of PURA, at 5q31, and PURB, at 7p13, in myelodysplastic syndrome and progression to acute myelogenous leukemia. AB - Deletions or monosomy of chromosomes 5 and 7 are frequently observed in myelodysplastic syndromes (MDS) and acute myelogenous leukemia (AML). In this study two genes, PURA and PURB, encoding functionally cooperative proteins in the Pur family, are localized to chromosome bands 5q31.1 and 7p13, respectively. One or both of these loci are shown to be hemizygously deleted in 60 MDS or AML patients using fluorescence in situ hybridization (FISH). High-resolution mapping of PURA localizes it approximately 1.1 Mb telomeric to the EGR-1 gene. Frequency of PURA deletion and segregation with EGR-1 indicate that PURA is within the most commonly deleted segment in myeloid disorders characterized by del(5)(q31). No mutations have been detected within the coding sequence of PURA. Concurrent deletions of PURA and PURB occur in MDS at a rate nearly 1.5-fold higher than statistically expected and in AML at a rate > 5-fold higher. This novel simultaneous deletion of two closely related gene family members may thus have consequences related to progression to AML. Pur alpha, an Rb-binding protein, has been implicated in cell cycle control and differentiation, and Pur alpha and Pur beta are reported to function as heterodimers. Alterations in these genes could affect a delicate balance critical in myeloid development. PMID- 11417484 TI - Secondary acute myelogenous leukemia and myelodysplasia without abnormalities of chromosome 11q23 following treatment of acute leukemia with topoisomerase II based chemotherapy. AB - Therapy-related MDS and AML are complications of intensive chemotherapy regimens. Traditionally, patients exposed to topoisomerase II inhibitors are reported to develop secondary AML with abnormalities of chromosome 11q23. We evaluated the long-term hematologic toxicity of topoisomerase II-intensive high-dose mitoxantrone-based chemotherapy in 163 newly diagnosed acute leukemia patients treated over an 8 year period. Nine (5.5%) patients developed new cytogenetic abnormalities. Four patients developed MDS with progression to AML, three patients developed new abnormalities at the time of relapse, and three patients (including one of the former patients) had changes that were not associated with hematologic disease. The abnormalities most frequently involved chromosomes 7q, 20q, 1q, and 13q. Despite the use of topoisomerase II-intensive treatment, no patient developed an abnormality involving chromosome 11q23. Spontaneous resolution of some changes and prolonged persistence of others in the absence of hematologic disease indicates that some cytogenetic changes are not sufficient to promote leukemogenesis. PMID- 11417485 TI - Meiotic origin of trisomy in neoplasms: evidence in a case of erythroleukaemia. AB - Trisomic cells in neoplasms may represent abnormal clones originated from a tissue-confined mosaicism, and arise therefore by a meiotic error. We report on a 16-month-old child with erythroleukaemia (AML-M6), whose marrow karyotype at onset was 48,XX,del(13)(q12q14),del(14)(q22q32),+21,+21. The parental origin of the supernumerary chromosomes 21 was investigated by comparing 10 polymorphic loci scattered along the whole chromosome on the patient's marrow and her parents' leukocytes. Three loci were informative for the presence of three alleles, two of which were of maternal origin; two further loci showed a maternal allele of higher intensity. Lymphocytes and skin fibroblasts showed a normal karyotype, and molecular analysis on leukocytes at remission, buccal smear and urinary sediment cells consistently showed only one maternal allele, whereas neonatal blood from Guthrie spot showed two maternal alleles as in the marrow. An accurate clinical re-evaluation confirmed a normal phenotype. Our results indicate that tetrasomy 21 arose from a marrow clone with trisomy 21 of meiotic origin. To the best of our knowledge, this is the first evidence that supernumerary chromosomes in neoplastic clones may in fact be present due to a meiotic error. This demonstrates that a tissue-confined constitutional mosaicism for a trisomy may indeed represent the first event in multistep carcinogenesis. PMID- 11417486 TI - Elevated vascular endothelial growth factor (VEGF) serum levels in idiopathic myelofibrosis. AB - An increase of angiogenesis has been shown in idiopathic myelofibrosis with myeloid metaplasia (MMM) by microvessel density count method but evaluation of circulating angiogenic factors is still incomplete. In 31 patients affected by MMM and in 12 healthy subjects we evaluated the serum levels of VEGF (vascular endothelial growth factor) and correlated VEGF with clinical and laboratory features of disease. We found that MMM patients had circulating VEGF concentrations much higher than controls (Median 1208 ng/ml vs 138 ng/ml, P < 0.0001). No correlation was found between VEGF and Hb, WBC, PLT, LDH, creatinine, bone marrow cellularity, fibrosis, splenomegaly, hepatomegaly, and therapy. However, in the subgroup of patients with a normal or low VEGF concentration, a direct correlation between VEGF and platelet count (r = 0.90, P = 0.002) was detected. Moreover, patients with a platelet count < 300 x 10(9)/l had VEGF serum levels lower than patients with a higher PLT count (median VEGF 864 vs 1557 pg/ml, P = 0.001). In six patients and in eight controls we also had the opportunity to measure VEGF in the plasma and we calculated that VEGF concentration was much higher in platelet-rich than in platelet-poor plasma and that platetets of MMM patients contained four times more VEGF than those of healthy controls. These results indicate that VEGF is overproduced in MMM, thus confirming an increased angiogenic activity. Platelets are probably a major source of VEGF in MMM but not the only one. PMID- 11417487 TI - Deletions of chromosome 13 in multiple myeloma identified by interphase FISH usually denote large deletions of the q arm or monosomy. AB - Deletions of the long arm of chromosome 13 (13q-) are observed in patients with multiple myeloma (MM), are rarely observed in the monoclonal gammopathy of undetermined significance (MGUS) and have been associated with a worsened prognosis in MM. However, no minimally deleted region in the 13q arm has been defined at 13q, and consequently no tumor suppressor genes have yet been identified that are important for disease pathogenesis. We attempted to characterize these chromosome 13q deletions at the molecular cytogenetic level. We studied 351 newly diagnosed patients, entered into the E9486/E9487 clinical study of the Eastern Cooperative Oncology Group. Fluorescent in situ hybridization (FISH) combined with immune fluorescent detection (cIg-FISH) of clonal plasma cells (PC) and cytomorphology were used to analyze interphase, bone marrow (BM) cell, cytospin slides. We simultaneously used DNA probes for the following locus specific probes (LSI); LSI 13 (Rb) and D13S319, which hybridize to 13q14. We subsequently studied distal deletions using the D13S25 probe (13q14.3) and a subtelomeric probe (13qSTP) for the 13q-arm (D13S327) in 40 cases with documented LSI 13 (Rb)/D13S319 deletion and 40 without deletion of these loci. Of 325 evaluable patients, we found 13q deletions in 176 (54%) using LSI 13 (Rb) and D13S319 probes. Of 40 patients with LSI 13 (Rb)/D13S319 deletions, 34 (85%) had coexistent deletion of both D13S25/13qSTP. These results indicate that chromosome 13 deletions in MM involve loss of most if not all of the 13q arm perhaps even indicating monosomy. In six cases the 13qSTP signal was conserved, but D13S25 was lost indicating large interstitial deletions involving 13q14. In 39 of the 40 cases without LSI 13 (Rb)/D13S319 deletions, the normal pattern of two pairs of signals was observed for D13S25/13qSTP. Deletions involving 13q14 are very common in MM as detected by cIg-FISH. These deletions appear to predominantly involve loss of large segments of the 13q arm or monosomy 13, and only occasionally represent an interstitial deletion. PMID- 11417488 TI - An atlas of chromosomes in hematological malignancies. Example: 11q23 and MLL partners. PMID- 11417489 TI - ABL-specific tyrosine kinase inhibitor, STI571 in vitro, affects Ph-positive acute lymphoblastic leukemia and chronic myelogenous leukemia in blastic crisis. PMID- 11417490 TI - Relationship between methylation of the p15 gene and ectopic expression of the EVI-1 gene in myelodysplastic syndromes (MDS). PMID- 11417491 TI - Short-term growth potential of cryopreserved B chronic lymphocytic leukemia cells in suspension and in semisolid medium. PMID- 11417492 TI - Triple-negative (CD3-/CD4-/CD8-) adult T cell leukemia/lymphoma, histologically presenting as CD30 (Ki-1)-positive anaplastic large cell lymphoma with clonal Epstein-Barr virus genome. PMID- 11417493 TI - Expression of two types of E2A-HLF fusion proteins in YCUB-2, a novel cell line established from B-lineage leukemia with t(17;19). PMID- 11417494 TI - Do transcriptionally silent BCR-ABL cells persist in CML patients in molecular remission after stem cell transplantation? PMID- 11417495 TI - Refinement of vaccine potency testing with the use of humane endpoints. PMID- 11417496 TI - Humane endpoints and cancer research. PMID- 11417497 TI - [The erosive potential of various oral care products compared to foodstuffs and beverages]. AB - The purpose of the present study was to investigate the erosive potential of various oral care products and to compare the results with those of various foodstuffs and beverages. Furthermore, the chosen method of surface hardness measurements was compared with scanning electron microscopy. Seven oral care products (toothpastes and mouth rinses), three foodstuffs and one beverage (as positive control) were tested for their erosive potential. After initial hardness testing 110 enamel specimens were distributed into 11 groups of 10 specimens each in such a way as to have a homogenous hardness distribution per group. The erosive potential of the products was tested by immersing the enamel specimens into solutions of the various products for 10 and 20 minutes. After the experimental period Knoop hardness was measured on the enamel specimens and the differences between before and after were calculated. The micromorphology of native enamel surfaces before and after immersion was determined by SEM on further enamel specimens treated identically for 20 minutes. Apple puree and orange juice produced the most marked losses of hardness after 10 and 20 minutes, respectively. Among the dental hygiene products only the fluoride-free Weleda toothpaste produced a significant reduction of hardness after 10 as well as after 20 minutes. With the exception of Elmex-Gel, all tested dental hygiene products led to a significant (p<0.05) increase of enamel hardness. The results of the hardness measurements corresponded with the micromorphological findings. It may, therefore, be concluded that the tested fluoride-containing oral care products do not exhibit any erosive potential which could be measured with the methodology used in this investigation. PMID- 11417498 TI - [The influence of different polishing systems on dental veneering materials]. AB - Experimental studies were conducted to investigate the efficacy of seven polishing systems (Dia Finish, Kohinoor, Dia Glace, Kenda, Shofu, UNOR, Opal) when used on seven ceramic materials (Empress, Vitadur Alpha, Celay, Cerec, Vita Titanium Porcelain, TiBond, Duceratin), in some cases also on resins (Visiogem, Spectrasit, composite, resin teeth), and on natural enamel. The surface quality of 100 specimens was examined with an optical profilometer and a scanning electron microscope and subsequently evaluated. Polishing the ceramic materials with Kohinoor or Dia Finish resulted in a very high surface polish.. Of the resins, Spectrasit yielded a very high polish when Dia Finish, Dia Glace or Opal were used. Using the tested polishing systems, surface polish results can be obtained intraorally on ceramic restorations that are at least equivalent to glaze-fired ceramic surfaces, if not better. This means that an additional glaze firing is no longer required when ceramic restorations have been modified by grinding. The Dia Finish, Dia Glace, and Kohinoor polishing systems are particularly recommended for finishing and polishing cemented ceramic or resin inlays. The results are similarly favorable when polishing the natural dental enamel, so an excellent homogenous transition from enamel to dual cement and ceramic restoration can be achieved during the final polishing step for cemented restorations. PMID- 11417499 TI - Re: Dietary fat, ketosis, and seizure resistance in rats on the ketogenic diet. PMID- 11417500 TI - MIMiC-based IMRT- part I. PMID- 11417501 TI - Dosimetry bibliography. PMID- 11417502 TI - Non-coplanar inverse planning IMRT using the MIMiC system: clinical significance in choice of 2-cm/1-cm mode and single couch vs. multiple couch angles. AB - The IMRT delivery process using MIMiC is extremely effective. In the hands of an experienced practitioner, MIMiC can provide excellent conformity to tumor volume with minimal damage to adjacent critical structures. However, the process is not completely automatic, even though it is an inverse planning process. The choice of machine treatment mode, 1- or 2-cm mode, and the choice of single couch angle vs. multiple, will yield quite different end results, with all prescription parameters being equal. The choice of mode and number of couch angles that yield excellent results of good coverage of the PTV with minimal hot spots and appropriate degree of normal tissue sparing still depends on the clinical experience of the planner. The size, shape, and total length of the PTV and those of the organs of interest dictate the choice. The authors demonstrated some clinical examples with comparative studies of the results of different choice of these parameters. In general, smaller beamlets with multiple couch angles give better PTV conformity, and critical organ avoidance. The down side of these choices are: (a) relatively more scattered hot spots within PTV, (b) longer time required to deliver the treatment, and (c) increased complexity of the process that could increase the chance of human error in treatment delivery. PMID- 11417503 TI - The University of California, Irvine experience with tomotherapy using the Peacock system. AB - Our institutional experience using the Peacock system for intensity-modulated radiation therapy (IMRT) is summarized. Over 100 patients were treated using this system, which is fitted to a Clinac 600C linac. Both cranial and extracranial lesions have been treated using this modality. Immobilization is achieved either with the Talon system for cranial sites or an Aquaplast cast. Target volumes up to 500 cm3 have been treated. Multiple lesions (up to 3) were treated in one setup. The range of dose/fractionation schemes used was 15 Gy/1 fx (radiosurgical treatment) - 80 Gy/40 fx. Dose validation studies were carried out using film and ion chamber dosimetry in a specially designed phantom. Optimal dose distributions were attainable using inverse treatment planning for IMRT delivery. These were found to encompass the target volumes accurately using dose validation phantom studies. Immobilization methods used were accurate to within 1 mm, as evidenced by daily portal films. IMRT using the Peacock system offers the advantage of delivery of conformal therapy to high doses safely and accurately. This provides the opportunity for dose escalation studies, retreatment of previously treated tumors, as well as treating multiple targets in one setup. The system may be fitted to a conventional linac without major modifications. PMID- 11417504 TI - Independent dose calculations for the PEACOCK System. AB - An independent dose calculation method has been developed to validate intensity modulated radiation therapy (IMRT) plans from the NOMOS PEACOCK System. After the plan is generated on the CORVUS planning system, the beam parameters are imported into an independent workstation. The beam parameters consist of intensity maps at each gantry angle and each arc position. In addition, CT scans of the patient are imported into the independent workstation to obtain the external contour of the patient. The coordinate system is defined relative to the alignment point chosen in the CORVUS plan. The independent calculation uses the pencil beam data viz tissue maximum ratio (TMR) and beam profiles for a single 1 x 0.8-cm beamlet formed by the NOMOS multileaf intensity-modulating collimator (MIMiC) leaf. The pencil beam data were measured for the 6-MV photon beam from Siemens PRIMUS linear accelerator using film dosimetry. The dose at a point is calculated using the depth and off-axis distance from a given pencil beam, corrected for its beam intensity. Isodose distributions are generated using the independent dose calculations and compared to the CORVUS plans. Isodose distributions show good agreement with the CORVUS plans for a number of clinical cases. The independent dose calculation algorithm is described in this paper. PMID- 11417505 TI - Where goest the Peacock? AB - Since the treatment of the first patient in 1994, the Peacock system has maintained its presence as the dominant method of intensity-modulated radiation therapy (IMRT) delivery. Currently in use at nearly 80 institutions, over 8000 patients have been treated using the system. Peacock treatments have been delivered to sites throughout the body, including CNS, head & neck, prostate, liver, kidney, lung, mediastinum, and extremities. IMRT, however, is a young and rapidly evolving treatment methodology. As institutions have explored new ways of improving radiation therapy with intensity-modulated techniques, the requirements for the Peacock system have also expanded. More sophisticated planning algorithms have been implemented to satisfy these new requirements, as well as better tools for treatment verification and quality assurance. In addition, new delivery techniques are being examined to improve the ability of IMRT to increase target volume doses while limiting organ-at-risk doses. One such technique, using helical tomotherapy (Peacock is an example of sequential tomotherapy), is currently being evaluated at one institution. Both techniques use narrow, modulated delivery beams. However, helical tomotherapy requires continuous movement of the couch during radiation, similar to helical CT. This work reviews the development of tomotherapy with the Peacock system. It then looks at current IMRT treatment techniques using tomotherapy, and how the field has broadened since the first treatments were delivered. Finally, it looks at the future of tomotherapy techniques, and how these techniques will adapt to the changing requirements for radiation therapy. PMID- 11417506 TI - NOMOS Peacock IMRT utilizing the Beak post collimation device. AB - Intensity-modulated radiation therapy (IMRT), an exciting recent development in the field of radiation therapy, is widely anticipated by many to make possible significant improvements in the quality of radiation treatments delivered to patients. The NOMOS Peacock method of delivery, often referred to as serial tomotherapy because of its "slice-wise" treatment of a tumor, has been used since 1994 to treat some 8000+ patients worldwide. This slice-wise method of treatment is known to produce extremely conformal dose distributions due to its ability to specifically match the dose distribution on each slice to the shape of the target volume on that same slice. Based on the belief of this institution, and the NOMOS Corporation, that an increase in the number of treatment slices into which the target is segmented would lead directly to an improvement in three-dimensional (3D) dose conformality, a joint effort was undertaken to develop a new MIMIC collimator treatment mode. Inherent to the original design of the NOMOS MIMIC binary multileaf collimator were 2 treatment modes: a 2-cm mode with a slice thickness of approximately 1.7 cm and a 1-cm mode with a slice thickness of approximately 0.85 cm. As a result of this collaborative effort, a new MIMIC treatment mode has been developed. The method employs a slit collimator, post collimation device known as the BEAK, enabling the treatment mode referred to as Beak Mode. The device imposes a distal redefinition of the slice thickness, or length, by effectively blocking the full retraction of the MIMIC vanes. The end result is a newly available slice thickness of approximately 4 mm, which is shown in this work to yield significant improvements in dose conformality for 2 representative patients. The comparative analysis of these 2 patient plans includes, in addition to a comparison of isodose distributions, an evaluation of dose-volume histogram (DVH) information, and a comparison of indices of conformality (CI) and homogeneity (HI). PMID- 11417507 TI - Comparative study between IMRT with NOMOS BEAK and linac-based radiosurgery in the treatment of intracranial lesions. AB - A comparative study was undertaken to examine intracranial irradiation using intensity-modulation radiation therapy (IMRT) and linear accelerator-based radiosurgery. The IMRT was examined using the Peacock system with a BEAK attachment. A clinical case involving a metastatic brain lesion, treated with 3 radiosurgery isocenters, was planned for IMRT. The radiosurgery was planned using the Leibinger planning system. The IMRT was planned using the CORVUS planning system. The CORVUS planning system uses an inverse planning algorithm, a recent development in radiotherapy. Isodose distributions and dose volume histograms were generated and compared. Analysis of the dosimetry shows that the dose conformity and homogeneity within the target using the RTOG guidelines are superior for IMRT. The advantages of IMRT using inverse planning system include the ease of planning and execution of treatment, especially for cases that involve concave targets that require multiple isocenters using radiosurgery. PMID- 11417508 TI - Commissioning of Peacock System for intensity-modulated radiation therapy. AB - The Peacock System was introduced to perform tomographic intensity-modulated radiation therapy (IMRT). Commissioning of the Peacock System included the alignment of the multileaf intensity-modulating collimator (MIMiC) to the beam axis, the alignment of the RTA device for immobilization, and checking the integrity of the CRANE for indexing the treatment couch. In addition, the secondary jaw settings, couch step size, and transmission through the leaves were determined. The dosimetric data required for the CORVUS planning system were divided into linear accelerator-specific and MIMiC-specific. The linear accelerator-specific dosimetric data were relative output in air, relative output in phantom, percent depth dose for a range of field sizes, and diagonal dose profiles for a large field size. The MIMiC-specific dosimetric data were the in plane and cross-plane dose profiles of a small and a large field size to derive the penumbra fit. For each treatment unit, the Beam Utility software requires the data be entered into the CORVUS planning system in modular forms. These modules were treatment unit information, angle definition, configuration, gantry and couch angles range, dosimetry, results, and verification plans. After the appropriate machine data were entered, CORVUS created a dose model. The dose model was used to create known simple dose distribution for evaluation using the verification tools of the CORVUS. The planned doses for phantoms were confirmed using an ion chamber for point dose measurement and film for relative dose measurement. The planning system calibration factor was initially set at 1.0 and will be changed after data on clinical cases are acquired. The treatment unit was released for clinical use after the approval icon was checked in the verification plans module. PMID- 11417509 TI - Basic concepts of CORVUS dose model. AB - Basic concepts of the dose model utilized in the CORVUS treatment planning system are reviewed. Following the Peacock delivery tool (MIMiC) by NOMOS Corporation, CORVUS "delivers" radiation to a patient by means of narrow x-ray beams (pencil beams), which are subject to lateral electronic disequilibrium. Dose data for such beams are difficult to obtain experimentally. Therefore, the CORVUS dose model uses analytically calculated (rather than experimentally measured) narrow beam dose data. The model is based on the idea that physical parameters necessary to calculate absorbed dose in narrow x-ray beams can be derived from measured broad-beam dose data. Calculation of central-axis and off-axis absorbed dose in narrow beams as well as a method of generating beam profiles are described. PMID- 11417510 TI - Immobilization devices for intensity-modulated radiation therapy (IMRT). AB - Three-dimensional conformal radiation therapy (3DCRT) and intensity-modulated radiation therapy (IMRT) plans show radiation dose distribution that is highly conformal to the target volume. The successful clinical implementation of these radiotherapy modalities requires precise positioning of the target to avoid a geographical miss. Effective reduction in target positional inaccuracies can be achieved with the proper use of immobilization devices. This paper reviews some of the immobilization devices that have been used and/or have the potential of being used for IMRT. The immobilization devices being reviewed include stereotactic frame, Talon system, thermoplastic molds, Alpha Cradles, and Vac-Lok system. The implementation of these devices at various anatomical sites is discussed. PMID- 11417511 TI - Abutment dosimetry for serial tomotherapy. AB - The dose distributions at the abutment region for serial tomotherapy are reviewed. While tomotherapy provides unparalleled dose distributions, precise couch motion and good patient immobilization are required because the dose in the abutment region changes by 25% for each millimeter of misalignment. The process of delivering intensity-modulated radiation therapy using sequentially delivered modulated arcs yields hot spots below and cold spots above the machine isocenter when arc angles of less than 360 degrees are used. The magnitude of the hot and cold spots increases significantly as the arc angle is reduced 180 degrees such as when limited by couch clearance restrictions. Placement of isocenter also significantly affects the dose heterogeneity in the abutment region, with the hot and cold spots increasing nearly linearly with off-axis distance in the vertical direction. Reduction of the magnitude of the abutment region dose heterogeneities is possible if helical delivery is provided by moving the couch during arc delivery. The dose heterogeneity can also be reduced by creating 2 treatment plans, each with slightly different abutment region positions, or by using multiple couch angles. PMID- 11417512 TI - Quality assurance procedures for the Peacock system. AB - The Peacock system is the product of technological innovations that are changing the practice of radiotherapy. It uses dynamic beam modulation technique and inverse planning algorithm, both of which are new methodologies, to perform intensity-modulation radiation therapy (IMRT). The quality assurance (QA) procedure established by Task Group No. 40 did not adequately consider these emerging modalities. A review of literature indicates that published articles on QA procedures concentrate primarily on the verification of dose delivered to phantom during commissioning of the system and dose delivered to phantom before treating patients. Absolute dose measurements using ion chambers and relative dose measurements using film dosimetry have been used to verify delivered doses. QA on equipment performance and equipment safety is limited. This paper will discuss QA on equipment performance, equipment safety, and patient setup reproducibility. PMID- 11417513 TI - Viewpoints to consider in the development of MLC and IMRT programs. AB - Operational and administrative considerations for the development and implementation of either multileaf collimation (MLC) or non-MLC-based intensity modulated radiation therapy (IMRT) will be discussed with respect to the impact that this will have on a radiation oncology department. The methodologies that are used in the development process will be reviewed in both the operational and administrative components. Operational issues addressing the potential clinical impact the department will experience, along with the beneficial outcome for the patients, will be major points in this review process. Administrative considerations generally focus on the financial impact that the department and hospital will have to deal with in determining cost benefits of this capital expenditure. This overview will focus on some key discussion points with the incorporation of MLC and IMRT into a radiation oncology department treatment program. PMID- 11417515 TI - Medicare+Choice. Though they're in the minority, some seniors seem to like managed care. PMID- 11417514 TI - Malpractice litigation. Plaintiffs lose more often--but when they win, they win big. PMID- 11417516 TI - Ash burns of the feet. PMID- 11417517 TI - An unusual burn caused by ignition of a floor-polishing product. PMID- 11417518 TI - Carsin H, et al. Cultured epithelial autografts in extensive burn coverage of severely traumatized patients: a five year single-center experience with 30 patients. Burns 2000;26:379-387. PMID- 11417519 TI - Relationship between mood disorders and coronary artery disease: possible role of the immune system and infection. PMID- 11417520 TI - The effect of bed rest on the diurnal variation in insulin resistance. PMID- 11417521 TI - Apolipoprotein E polymorphism and C-reactive protein in dyslipidemic middle-aged men. PMID- 11417522 TI - Light-to-moderate alcohol consumption, risk factor profile and early atherosclerosis: the RIAD study. PMID- 11417523 TI - The mutation APOB-100 R3500Q in Eastern Europe. PMID- 11417524 TI - HIV in Europe--a growing threat. PMID- 11417525 TI - Contact irritation provoking Hailey-hailey disease. PMID- 11417526 TI - Occupational contact allergy to tetrazepam. PMID- 11417527 TI - Contact sensitization to chlorproethazine can induce persistent light reaction and cross-photoreactions to other phenothiazines. PMID- 11417528 TI - Occupational contact dermatitis due to the epoxy hardener m-xylylenediamine. PMID- 11417529 TI - Sir Hans A. Krebs Centenary Lecture: cancer and clinical targeting. PMID- 11417530 TI - Pitfalls and uses of medical databases in research. PMID- 11417531 TI - A palliative accelerated irradiation regimen for advanced non-small-cell lung cancer vs. conventionally fractionated 60 Gy: results of a randomized equivalence study: regarding Nestle et al. IJROBP 2000; 48:95-103. PMID- 11417532 TI - Moving brachytherapy from art to science. PMID- 11417533 TI - Is there a problem in the application of the current tumor control probability model to (prostate) clinical data? PMID- 11417534 TI - Caregiver toxicity from transdermal fentanyl. PMID- 11417535 TI - Clinical problems with transdermal fentanyl titration from 25 to 50 microg/hr. PMID- 11417536 TI - The great unifier? AMA's plan for a 'core organization' meets specialty-group resistance. PMID- 11417537 TI - Effectiveness of medical therapy alone compared with coronary angioplasty or left internal mammary artery bypass for single-vessel coronary artery disease. PMID- 11417538 TI - Mentor for MENTOR. PMID- 11417539 TI - Cardiosource is a good source for definition of trial acronyms. PMID- 11417540 TI - Medical errors. Analysis of adverse events must result in improvements in care. PMID- 11417541 TI - Medical errors. Organisational ethos is important. PMID- 11417542 TI - Medical errors. System for reporting errors is not highest priority to decrease errors. PMID- 11417543 TI - Medical errors. Media tend to link error with blame. PMID- 11417544 TI - Medical errors. Perhaps blame-free culture is needed in NHS to reduce errors. PMID- 11417545 TI - Medical errors. Terminology of "error" is important. PMID- 11417547 TI - Medical errors. Errors can have their uses. PMID- 11417548 TI - Medical errors. Courses on crisis avoidance and resource management are available. PMID- 11417549 TI - Medical errors. Different formulations of drugs often look confusingly similar. PMID- 11417550 TI - Medical errors. Appropriate training should avoid accidental intrathecal injection of vincristine. PMID- 11417551 TI - Medical errors. Dosage nomenclature of bleomycin needs to be standardised to avoid errors. PMID- 11417552 TI - Medical errors. Chemotherapy regimens have been formalised into protocols in British Columbia. PMID- 11417553 TI - Medical errors. Medical schools can teach safe drug prescribing and administration. PMID- 11417555 TI - Medical errors. Medical profession must take drug errors seriously. PMID- 11417556 TI - Medical errors. Consent is outdated concept. PMID- 11417557 TI - Adverse events in British hospitals. "Errors meetings" in radiology did not identify errors leading to complaints and litigation. PMID- 11417558 TI - Adverse events in British hospitals. Preventive strategies, not epidemiological studies, are needed. PMID- 11417559 TI - Adverse events in British hospitals. Adverse events may occur whatever course of action is pursued. PMID- 11417560 TI - Adverse events in British hospitals. Hospital acquired infections consume bed days and resources. PMID- 11417561 TI - Adverse events in British hospitals. Threshold used for determining adverse events is important. PMID- 11417562 TI - Adverse events in British hospitals. Retrospective case record analysis has been superseded. PMID- 11417563 TI - Adverse events in British hospitals. Expanded definition of adverse events is needed. PMID- 11417564 TI - [5-HT1B serotonin receptors and antidepressant effects of selective serotonin reuptake inhibitors ]. AB - We used knockout mice and receptor antagonist strategies to investigate the contribution of the serotonin (5-hydroxytryptamine, 5-HT) 5-HT1B receptor subtype in mediating the effects of selective serotonin reuptake inhibitors (SSRIs). Using in vivo intracerebral microdialysis in awake mice, we show that a single systemic administration of paroxetine (1 or 5 mg/kg, i.p.) increased extracellular serotonin levels [5-HT]ext in the ventral hippocampus and frontal cortex of wild-type and mutant mice. However, in the ventral hippocampus, paroxetine at the two doses studied induced a larger increase in [5-HT]ext in knockout than in wild-type mice. In the frontal cortex, the effect of paroxetine was larger in mutants than in wild-type mice at the 1 mg/kg dose but not at 5 mg/kg. In addition, either the absence of the 5-HT1B receptor or its blockade with the mixed 5-HT1B/1D receptor antagonist, GR 127935, potentiates the effect of a single administration of paroxetine on [5-HT]ext more in the ventral hippocampus than in the frontal cortex. Furthermore, we demonstrate that SSRIs decrease immobility in the forced swimming test; this effect is absent in 5-HT1B knockout mice and blocked by GR 127935 in wild-type suggesting therefore that activation of 5-HT1B receptors mediate the antidepressant-like effects of SSRIs. Taken together these data demonstrate that 5-HT1B autoreceptors appear to limit the effects of SSRI on dialysate 5-HT levels particularly in the hippocampus while presynaptic 5-HT1B heteroreceptors are likely to be required for the antidepressant activity of SSRIs. PMID- 11417565 TI - Clarification--Drug risk in patients with glaucoma. PMID- 11417566 TI - Rhabdomyolysis due to cerivastatin monotherapy. PMID- 11417567 TI - Recurrent aortic root hematograft endocarditis related to indwelling hemodialysis catheters. PMID- 11417568 TI - Extreme hyponatremia of 87 mmol/l without neurologic complications in a severely hypovolemic patient. PMID- 11417569 TI - Acute pancreatitis as the first manifestation of pancreatic Burkitt's lymphoma in a patient infected by the human immunodeficiency virus. PMID- 11417570 TI - Assessing adjuvant breast cancer therapy benefit. PMID- 11417571 TI - Neonatal salt intake and blood pressure. PMID- 11417572 TI - Neonatal salt intake and blood pressure. PMID- 11417573 TI - Neonatal salt intake and blood pressure. PMID- 11417574 TI - Salicylates, nitric oxide, malaria, and Reye's syndrome. PMID- 11417575 TI - On-demand relief treatment for asthma. PMID- 11417576 TI - On-demand relief treatment for asthma. PMID- 11417577 TI - Dopamine D2 receptor and obesity. PMID- 11417578 TI - Dopamine D2 receptor and obesity. PMID- 11417579 TI - Consent for fetal necropsy. PMID- 11417580 TI - Cuba's free medical education for US students. PMID- 11417581 TI - Cuba's free medical education for US students. PMID- 11417582 TI - Cell-wall-deficient bacteria. PMID- 11417583 TI - Cell-wall-deficient bacteria. PMID- 11417584 TI - Treatment of menorrhagia. PMID- 11417585 TI - Statins and fractures. PMID- 11417586 TI - Worms that cause lumps in the mouth. PMID- 11417587 TI - Treatment of amblyopic eyes. PMID- 11417588 TI - Treatment of amblyopic eyes. PMID- 11417589 TI - Does myalgic encephalomyelitis exist? PMID- 11417590 TI - Does myalgic encephalomyelitis exist? PMID- 11417591 TI - Does myalgic encephalomyelitis exist? PMID- 11417592 TI - Declining enrollment in podiatric medical school market forces at work and an opportunity to seize. PMID- 11417593 TI - Biomechanics of the first ray. Part III. Consequences of Lapidus arthrodesis on peroneus longus function: a three-dimensional kinematic analysis in a cadaver model. AB - It has long been proposed that first metatarsocuneiform joint (FMCJ) arthrodesis, also known as Lapidus arthrodesis, can realign the first ray and permanently lock the FMCJ to control hypermobility. Left unanswered is the functional consequence of peroneus longus (PL) after such a procedure. In this study, the effects of PL on the medial column of the foot before and after metatarsocuneiform arthrodesis were evaluated. Seven fresh-frozen cadaver specimens with an intact foot and ankle were mounted on a custom-made acrylic frame and loaded to 400 N while midstance motor function was simulated with pneumatic actuators. Three dimensional radiowave tracking transducers were attached to the first metatarsal, medial cuneiform, navicular, and talus to measure osseous movements while tensile loads of 0% to 100% of PL predicted force was applied. Simulated arthrodesis of the metatarsocuneiform joint and then additionally the intercuneiform 1-2 joint was achieved with titanium pins and then retested to determine any change in effect from PL. Significant frontal plane eversion of the medial cuneiform (p = .016) and dorsiflexion of the talus (p = .045) occurred after Lapidus arthrodesis was achieved. This suggests that arthrodesis at the first metatarsocuneiform joint increases the efficiency of PL stabilizing action on the medial column. PMID- 11417594 TI - Magnetic resonance and cadaveric findings of the "watershed band" of the achilles tendon. AB - A review of 63 magnetic resonance images of 86 ankles was performed. (There were 23 bilateral studies.) A contiguous fascial band from the deep posterior compartment to the Achilles tendon watershed region was noted in 83 ankles. This fascial band enveloped the watershed region, coursing from the flexor (lacinate) ligament medially, to the peroneal retinaculum laterally. This band was absent in three patients who were scanned, two of whom sustained a complete Achilles tendon rupture, while the third had previously undergone peritenolysis. Forty cadaveric specimens with no known Achilles tendon pathology were evaluated for presence of this structure. An organized fascial band existed in all of these specimens. This previously undescribed structure is termed the "watershed band" and may have surgical significance. PMID- 11417595 TI - First metatarsophalangeal arthrodesis for treatment of hallux rigidus: a retrospective study. AB - This was a retrospective study of 17 patients (21 operated feet) treated with first metatarsophalangeal arthrodesis for hallux rigidus of varying severity levels. Patients were evaluated according to a modified American Orthopedic Foot and Ankle Society hallux metatarsophalangeal-interphalangeal rating scale containing 60 subjective and 30 objective points. At an average follow-up time of 28.1 months, patients demonstrated a mean subjective improvement from 21.2 to 47.8/60, with mean total scores improving from 39.1 to 75.6/90. A comparison of pre- and postoperative radiographic angular measurements demonstrated a significant improvement (p = .001) in intermetatarsal, first metatarsal declination, and lateral talo-first metatarsal angles. Significant positive correlations were found between subjective results and patient age (p = .05) and the preoperative lateral talo-first metatarsal angles (p = .001). PMID- 11417596 TI - The Mitchell bunionectomy: a prospective study of 60 consecutive cases utilizing single K-wire fixation. AB - The authors prospectively evaluated 45 patients (60 feet) affected by hallux valgus and treated with a distal metatarsal osteotomy. The surgical procedure consisted of a modified Mitchell osteotomy, in which fixation was achieved with a Kirschner wire that was driven into the proximal osteotomy fragment and buttressed the distal one. Early weightbearing was allowed without a cast. Follow up averaged 25 months. The mean American Orthopedic Foot and Ankle Society clinical hallux score increased from 44.6/100 preoperatively to 83.2/100. Radiographic evaluation showed that mean metatarsophalangeal and intermetatarsal angles decreased respectively from 31.7 degrees to 16.9 degrees, and from 15.4 degrees to 8.6 degrees. Short-term loss of correction occurred in three cases (4%). Six feet (10%) had unrelieved metatarsalgia that was related to excessive shortening of the first metatarsal and/or inappropriate orientation of the metatarsal head. Stabilization of the Mitchell osteotomy with a Kirschner wire proved safe and effective for the surgical correction of mild to moderate hallux valgus. PMID- 11417597 TI - Subtalar joint arthroscopy for sinus tarsi syndrome: a review of 29 cases. AB - Twenty-nine consecutive patients who underwent diagnostic or therapeutic subtalar joint arthroscopy for sinus tarsi syndrome were retrospectively reviewed. The mean length of follow-up was 18 months. There was a history of trauma in 86% of the patients, with an inversion sprain being the most common predisposing injury (63%). All patients had a primary preoperative diagnosis of sinus tarsi syndrome. Magnetic resonance imaging was useful in identifying subtalar joint chronic synovitis and/or fibrosis in all 26 patients who were imaged. Subtalar joint synovectomy was the most common procedure performed. Twelve patients had 15 additional operative procedures. One patient required an arthrotomy secondary to arthrofibrosis. There were no postoperative complications. The mean return to full activity was 4 months. The mean postoperative AOFAS Ankle-Hindfoot Scale score was 85 points. Subtalar joint arthroscopy has proven to be a relatively safe and effective diagnostic and therapeutic technique in the management of sinus tarsi syndrome. PMID- 11417598 TI - Reconstruction of skin and tendon defects from wound complications after Achilles tendon rupture. AB - Four patients who developed combined tendon and overlying skin defects following operative repair of ruptured Achilles tendon were presented. Three patients had an infected wound. The average interval from the first operation for repairing the ruptured Achilles tendon and the reconstructive procedure was 46.2 days (range, 5-65 days). All patients were treated with a one-stage operation including radical debridement, reconstruction of the Achilles tendon defects using vascularized or nonvascularized tendon grafts, transfer of peroneus brevis for augmentation, and skin coverage with a free flap. The patients recovered uneventfully. The average follow-up period was 39.2 months (range, 18-79 months). In all patients, an evaluation of the clinical outcome, the performance of the calf muscles using a computerized dynamometer, and structural changes of the reconstructed Achilles tendon using magnetic resonance (MR) imaging were made. The clinical outcome was excellent in three patients and good in one. In isokinetic testing (Cybex-Norm), strength was found to be normal in one patient and abnormal in three patients. MR images revealed an intratendinous area of homogenous and normal intensity signal, and a significant increase in thickness and width in all levels of the reconstructed Achilles tendon. The authors conclude that it is possible to obtain satisfactory function in patients with complex wounds in the region of the Achilles tendon. PMID- 11417599 TI - Treatment of congenital convex pes valgus with navicular excision and subtalar arthroereisis: a case study. AB - This article presents a case study of a 4-year-old female with a delayed diagnosis of congenital convex pes valgus. The severe soft-tissue contractures and osseous abnormalities in this case necessitated excision of the navicular and subtalar arthroereisis in addition to standard soft-tissue releases in order to achieve and maintain adequate reduction. A review of the literature and a discussion of these adjunctive techniques is provided to examine the concepts relevant to treatment of this condition. PMID- 11417600 TI - Autonomic dysreflexia and foot and ankle surgery. AB - Autonomic dysreflexia is a syndrome of massive imbalance of reflex sympathetic discharge occurring in patients with spinal cord injury with a lesion above the splanchnic outflow (Thoracic 6). Autonomic dysreflexia is characterized by a sudden and severe rise in blood pressure and is potentially life threatening. Because the onset of this entity is rapid and the potential morbidity is severe, it is important for those caring for spinal cord injury patients to be aware of this syndrome. The paper presents a review of the literature, and familiarizes one with the diagnosis, pathophysiology, and treatment. Two illustrative case reports are also presented. PMID- 11417601 TI - The calcaneal scarf osteotomy: operative technique. PMID- 11417602 TI - New trends in the etiopathogenesis of diabetic peripheral neuropathy. AB - Neuropathy is well recognized as a major complication of insulin-dependent diabetes mellitus in adults, resulting in significant morbidity and possibly an increased mortality. Both the peripheral and autonomic nervous systems can be involved, and adolescents with diabetes can show early evidence of neuropathy. The pathogenesis of diabetic neuropathy remains unclear but is thought to involve various mechanisms. This complication can be traced to the metabolic effects of hyperglycemia and/or other effects of insulin deficiency on the various constituents of the peripheral nerve. The polyol pathway and/or nonenzymatic glycation affecting one or more cell types in the multicellular constituents of the peripheral nerve appear likely to have an inciting role. The role of other factors, such as possible direct neurotrophic effects of insulin and insulin related growth factors, seems to be relevant. PMID- 11417603 TI - Risperidone monotherapy in preschool children with pervasive developmental disorders. AB - The aim of this preliminary study was to examine the short-term efficacy and safety of the atypical antipsychotic risperidone in preschool autistic children. The sample consisted of 10 subjects (7 males and 3 females) aged 3 9/12 to 6 6/12 years (mean age 4.7 years). A 16-week open-label trial with risperidone monotherapy was initiated at a starting dose of 0.25 mg daily and was increased to a maximum dose of 0.50 mg (0.027 mg/kg daily). Outcome measures were the Childhood Autism Rating Scale, the Children's Psychiatric Rating Scale, Clinical Global Impression (improvement score), and the Children's Global Assessment of Functioning. Two subjects did not complete the trial because of side effects (tachycardia and flushes, fever and hyporexia). After the 16-week treatment, data from the eight children who completed the trial indicated a modest improvement in the Childhood Autism Rating Scale total score, Children's Psychiatric Rating Scale total score, and Children's Global Assessment of Functioning. According to the Clinical Global Impression, the global improvement score for four subjects was much improved or very much improved; the score for the other four children was minimally improved. None of the children exhibited behavioral deterioration. The side effects in the eight children were not severe. PMID- 11417604 TI - Topography of cerebral white-matter disease of prematurity studied prospectively in 1607 very-low-birthweight infants. AB - The objective of this study was to evaluate to what extent (1) the characteristics of localization, distribution, and size of echodense and echolucent abnormalities enable individuals to be designated as having either periventricular hemorrhagic infarction or periventricular leukomalacia and (2) the characteristics of periventricular hemorrhagic infarction and periventricular leukomalacia are independent occurrences. The population for this study consisted of 1607 infants with birthweights of 500 to 1500 g, born between January 1991 and December 1993, who had at least one cranial ultrasound scan read independently by at least two ultrasonographers. The ultrasound data collection form diagrammed six standard coronal views. The cerebrum was divided into 17 zones in each hemisphere. All abnormalities were described as being echodense or echolucent and were classified on the basis of their size, laterality, location, and evolution. Eight percent (134/1607) of infants had at least one white-matter abnormality. The prevalence of white-matter disease decreased with increasing gestational age. Most abnormalities were small or medium sized and unilateral; only large echodensities tended to be bilateral and asymmetric. Large abnormalities, whether echodense or echolucent, were more likely than smaller abnormalities to be widespread, and the extent of cerebral involvement was independent of whether abnormalities were unilateral or bilateral. Large abnormalities were relatively more likely than small abnormalities to involve anterior planes. Small abnormalities, whether echodense or echolucent, or whether unilateral or bilateral, preferentially occurred near the trigone. Using the characteristics of location, size, and laterality/symmetry, we were able to allocate only 53% of infants with white-matter abnormalities to periventricular hemorrhagic infarction or periventricular leukomalacia. Assuming that periventricular leukomalacia and periventricular hemorrhagic infarction are independent and do not share risk factors, and that each occurs in approximately 5% of infants, we would have expected 0.25%, or about 4 individuals, to have abnormalities with characteristics of both periventricular leukomalacia and periventricular hemorrhagic infarction, whereas we found 63 such infants. Most infants with white matter disease could not be clearly designated as having periventricular hemorrhagic infarction or periventricular leukomalacia only. Periventricular hemorrhagic infarction contributes to the risk of periventricular leukomalacia occurrence, or the two sorts of abnormalities share common risk antecedent factors. The descriptive term echodense or echolucent and the generic term white matter disease of prematurity should be used instead of periventricular leukomalacia or periventricular hemorrhagic infarction when referring to sonographically defined white-matter abnormalities. PMID- 11417605 TI - Neurologic course of congenital disorders of glycosylation. AB - Congenital disorders of glycosylation, formerly called carbohydrate-deficient glycoprotein syndrome, may present in infancy with slowly progressive neurologic deficits including cognitive impairment, ataxia, pigmentary retinal degeneration, and neuropathy. The metabolic defect is in N-linked oligosaccharide synthesis, and diagnosis is made by a serum transferrin isoelectric focusing. We reviewed the neurologic course of 10 children with congenital disorders of glycosylation (ages 13 months to 7 years). All had severe developmental delay and ataxia; none walked unassisted, and the highest level of communication was simple sign language in one patient. Five of 10 children had seizures (absence, complex partial, tonic clonic). Only one patient has had strokelike episodes, despite reports that they are common in this population. The underlying basis of these episodes has been hypothesized to be coagulopathy due to dysfunctional, incorrectly glycosylated coagulation factors. This 5-year-old patient with congenital disorders of glycosylation type Ia had two strokelike episodes, with evolving hemiparesis over 5 to 6 days' duration, followed by focal tonic-clonic seizures. Coagulation studies were normal. Electroencephalography showed transient hemispheric polymorphous delta-range slowing and suppression. Magnetic resonance imaging revealed corresponding cortical swelling. Magnetic resonance angiography was normal. Magnetic resonance spectroscopy revealed a decrease in the N-acetylaspartate peak, suggesting neuronal loss, with normal lactate peak. The neuroradiologic data do not support a thrombotic, embolic, or hemorrhagic basis for strokelike episodes in carbohydrate-deficient glycoprotein syndrome; other mechanisms must be considered. PMID- 11417606 TI - Langerhans' cell histiocytosis presenting as intracranial hypertension. AB - Benign intracranial hypertension is known to be associated with obesity, endocrine abnormalities, various medications, and cerebral venous sinus thrombosis. We report a patient presenting with headaches and vomiting attributed to benign intracranial hypertension. The diagnostic work-up revealed Langerhans' cell histiocytosis of the occipital bone. There was no evidence for cerebral vein thrombosis by cranial computed tomography scan, Doppler ultrasonography, planar and single photon emission computed tomography technetium 99m-labelled red blood cell scintigraphy, and magnetic resonance angiography. Excision of the occipital bone lesion and a short course of acetazolamide and prednisone were curative. We hypothesize that cytokines secreted by the tumor were responsible for the development of intracranial hypertension. PMID- 11417607 TI - Interleukin-1beta, interleukin-1 receptor antagonist levels in patients with subacute sclerosing panencephalitis and the effects of different treatment protocols. AB - Subacute sclerosing panencephalitis is a rare progressive inflammatory disease of the central nervous system caused by a persistent aberrant measles virus infection. Cytokines are polypeptides that regulate immune responses and inflammatory reactions. Interleukin-1beta has been implicated as a central mediator of tissue damage and destruction in a number of central nervous system diseases. Interleukin-1 receptor antagonist could function as an important anti inflammatory cytokine. We studied interleukin-1beta and interleukin-1 receptor antagonist levels in the cerebrospinal fluids of patients with subacute sclerosing panencephalitis and evaluated the effects of different treatment protocols on these cytokines. Interleukin-1beta and interleukin-1 receptor antagonist levels were measured in 15 patients who had a recent diagnosis of subacute sclerosing panencephalitis (group 1), 6 patients who had been treated with isoprinosine (group 2), 5 patients with intraventricular interferon-alpha (group 3), and 6 patients with interferon-beta (group 4). The results were compared within the groups and also with the results of 10 patients with other neurologic disease (group 5). The interleukin-1beta concentrations in cerebrospinal fluid and sera were all below the detection limits (3.9 pg/mL). Interleukin-1 receptor antagonist levels were not statistically different, except for the group treated with intraventricular interferon-alpha. Interleukin-1 receptor antagonist levels were 170 +/- 52, 175 +/- 58, 1605 +/- 518, 77.5 +/- 24, and 108 +/- 18 pg/mL in groups 1 to 5, respectively. Interleukin-1 receptor antagonist levels and cerebrospinal fluid serum ratios were significantly increased during interferon-alpha treatment. In conclusion, interleukin-1 and interleukin-1 receptor antagonist levels were not elevated in the patients with subacute sclerosing panencephalitis. The only treatment protocol that affects interleukin-1 receptor antagonist levels in cerebrospinal fluid was intraventricular interferon-alpha. Further studies on higher numbers of patients may better document the immunologic status of patients with subacute sclerosing panencephalitis and the effects of different treatment modes. PMID- 11417608 TI - Brain volume in autism. AB - Increased brain size has been observed in individuals with autism with a wide range of cognitive functioning. The purpose of this investigation was to obtain measurements of the brain volume in a sample of nonmentally retarded autistic individuals. Magnetic resonance imaging scans from 16 nonmentally retarded individuals with autism and 19 male volunteer comparison subjects were obtained and the following structures were measured: third, fourth, and lateral ventricles and intracranial and cerebral volumes. Mean cerebral and third ventricle volumes in the autistic subjects were significantly greater than in the controls when adjusted for intracranial volume. No other significant results were found. Our finding of increased brain volume in autism is consistent with previous reports in the literature. Additional longitudinal neuroimaging and, more importantly, neuropathologic studies are warranted to provide a better understanding of the complexities underlying increased brain size in autism. PMID- 11417609 TI - Changes in skeletal maturation and mineralization in children with cerebral palsy and evaluation of related factors. AB - Sixty-nine (28 females, 41 males) children with spastic cerebral palsy and 26 (13 females, 13 males) healthy children were included in the study. Total- and partial-body bone mineral content and bone mineral density values of patient and control subjects were measured by dual-energy x-ray absorptiometry. Left hand and wrist radiographs of all patients and right hand and wrist radiographs of 39 randomly selected patients were taken, and the bone ages of all radiographs were determined. In both female and male tetraplegics, bone mineralization values of lower extremities, where the mobility disorder and effects of absence of weight bearing activity were maximal, were lower than those of controls and hemiplegics (P < .05). In 47 (68%) patients, left-side bone age values were below normal ranges for their ages, and the difference was statistically significant (P < .01). Our results indicate that motor function handicap affects skeletal mineralization adversely, and skeletal maturation is frequently delayed in children with cerebral palsy. We speculated that this delay might be a result of disrupted embryologic skeletal development due to hypoxic attack, which also causes the disease. PMID- 11417610 TI - Long-term follow-up of an epidemiologically defined cohort of patients with Tourette syndrome. AB - The goal of this study was to collect prospective longitudinal information on the development of an epidemiologically defined cohort of patients with Tourette syndrome. These data may improve prognostic understanding of the condition. This information will also be important for specification of an adult phenotype for genetic marker studies. A prospective longitudinal cohort study was conducted. Fifty-four of 73 patients from our 1986 prevalence study of Tourette syndrome in North Dakota school-aged children were eligible for inclusion. The subjects were diagnosed in 1984 and 1985. We were able to interview 39 of 54 eligible patients for 507 person-years of follow-up. For the cohort, tic severity declined by 59%, global assessment of functioning improved by 50%, and the average number of comorbidities decreased by 42%. Forty-four percent of patients were essentially symptom free at follow-up. Only 22% were on medication as adults. Tourette syndrome is a developmental neuropsychiatric disorder with a long-term course that is favorable for most patients. Males demonstrated substantially more variability in improvement but overall demonstrated more improvement than females. PMID- 11417611 TI - Evidence for apoptosis in the fetal Down syndrome brain. AB - In Down syndrome, enhanced apoptosis (programmed cell death) may play a role in the pathogenesis of characteristic early mental retardation and precocious neurodegeneration of Alzheimer type. Various apoptosis-associated proteins (Bax, Bcl-2, Fas, p53, Hsp70, neuronal apoptosis inhibitory protein-like immunoreactivity) were investigated in four different cortical regions and the cerebellum of one fetal Down syndrome (35 weeks' gestation) postmortem brain sample compared with a control brain sample. The most impressive finding was an at least fivefold elevation of Bax protein together with decreased Bcl-2 values in all Down syndrome cerebral regions investigated. In addition, antiapoptotic, presumably caspase-inhibitory, principles like heat shock protein 70 and neuronal apoptosis inhibitory protein were also reduced. Whereas Fas protein, an important member of receptor-mediated apoptosis, was inconsistently altered, a rather surprising finding was reduced proapoptotic, regulatory protein p53 in four of five regions. The findings are in good agreement with the proposed role of the Bcl-2 protein family in regulating developmental (naturally occurring) apoptotic neuronal death and further suggest that developmental apoptosis may be inappropriately commandeered by so far undefined pathologic processes in Down syndrome. PMID- 11417612 TI - Seizure recurrence in children with focal seizures and single small enhancing computed tomographic lesions: prognostic factors on long-term follow-up. AB - Single small enhancing computed tomographic (CT) lesions are common in children with focal seizures. There is a paucity of information regarding their long-term outcome and prognostic factors for seizure recurrence. The objective of this work was to study the frequency of seizure recurrence in children with single small enhancing computed tomographic lesions and to identify prognostic factors, if any, for seizure recurrence. A prospective long-term follow-up was conducted at the Advanced Pediatric Center, Postgraduate Institute of Medical Education and Research, an urban tertiary care teaching hospital. Sixty-three children between 2 and 12 years of age with focal seizures for less than 3 months and single small enhancing computed tomographic lesions were enrolled in a randomized, double blind, placebo-controlled trial of albendazole therapy and followed up for 4 years. On long-term follow-up, the albendazole and placebo groups were left with 29 and 28 children, respectively. After several months of seizure-free period, antiepileptic drug was tapered off. Children with relapse underwent magnetic resonance imaging examination. All children were followed up for at least 18 months after stopping of the antiepileptic drug. Seizure recurrence was seen in three children each in both groups, after a mean interval of 6.4 weeks after stopping the antiepileptic drug. Magnetic resonance imaging revealed persistent chronic granuloma in 2 and calcified granuloma in 4 children. Residual lesions were significantly correlated with seizure recurrence. In children whose lesions completely disappeared, no seizure recurrence was seen even during shorter periods of antiepileptic drug treatment. Seizure recurrence was seen in a small number of children with focal seizures and single small enhancing computed tomographic lesions. It appears to be related to either a persistent or a calcified lesion. PMID- 11417613 TI - Internal carotid artery thrombus: an underdiagnosed source of brain emboli in neonates? AB - We report a full-term neonate with a left middle cerebral artery infarct, narrowing of the internal carotid artery detected by magnetic resonance angiography and B-mode ultrasonography, and a large thrombus at the origin of the internal carotid artery detected by B-mode ultrasonography. Internal carotid arterial thrombus is seldom considered the source of middle cerebral arterial embolus in neonates. We suggest that B-mode ultrasonography of the carotid artery be included in the diagnostic evaluation of middle cerebral artery infarcts in neonates. PMID- 11417614 TI - Successful withdrawal from analgesic abuse in a group of youngsters with chronic daily headache. AB - Abuse of ergotamine and analgesics is common in adults. It coexists with headache and can also induce headaches. Ten to 15% of patients attending headache clinics and 1% of the general population suffer from chronic daily headache due to medication misuse. Indeed, this phenomenon was recently regarded as an epidemic. Nonetheless, analgesic-induced headache in children and adolescents was first reported in 1998. We report on our experience with children and adolescents with daily or almost-daily headache concomitant with daily or almost-daily analgesic intake. Over a period of 3 years, we evaluated 26 children (19 girls and 7 boys) with chronic daily or near-daily headache related to daily analgesic intake. The mean age of the group was 14.2 years (range, 12-18), and the mean headache history duration was 1.6 years (range, 3 months to 4 1/2 years). The mean number of headache days per month was 28.1 (range, 19-31). All patients had no history of migraine prior to the chronic headache phase according to the International Headache Society criteria. They were using at least one dose of analgesic drug for each headache, whereas 16 were using analgesic drugs daily. The weekly analgesic intake averaged 28.1 tablets (range, 19-41). The majority abused simple analgesics. Twenty-one took acetaminophen alone. Five took a combination; four took a compound containing acetaminophen, caffeine, and codeine; and the fifth patient took a compound containing aspirin, caffeine, and codeine. All patients were informed about the phenomenon of medication-induced headache and were encouraged to achieve drug withdrawal. Withdrawal led to complete cessation of all headaches in 20 patients. In 5 patients, the daily headache resolved; however, they suffered from intermittent episodic migraine attacks, which were frequent enough in 3 to initiate prophylactic medication. One adolescent continued to have daily headache. Analgesic-induced headache does occur in adolescents. Successful withdrawal from the offending analgesics was achieved without hospitalization or significant interference with daily life and with complete disappearance of the induced chronic daily headache in 25 of 26 patients. PMID- 11417615 TI - Three new patients with congenital unilateral facial nerve palsy due to chromosome 22q11 deletion. AB - We report three unrelated patients with congenital facial nerve palsy and chromosome 22q11 deletion, a condition hitherto poorly recognized. In the first case, facial palsy was associated with aortic coarctation, ductus arteriosus, and ostium secundum atrial septal defect. In the second case, facial palsy was associated with ostium secundum atrial septal defect, obstruction of the ureteropelvic junction, double ureteropelvic-calicial system, and distal metaphyseal widening of the forearm and leg bones. In both cases, facial palsy was the presenting feature. In the third case, an ostium secundum atrial septal defect was also present, but involvement of cranial nerves III, VI, and VIII, in addition to hypoplastic structures of cerebellar and cerebral peduncles, were the predominant features. There were no inherited deletions within chromosome band 22q11 and the de novo deletions detected in each case belonged to the paternally derived chromosome 22. Association of facial nerve palsy and congenital heart disease versus cardiofacial syndrome are different only on clinical grounds, so both conditions can be genetically identical and form part of the spectrum of defects associated with chromosome 22q11 deletions. We recommend investigation for chromosome 22q11 deletions in patients with complete nerve facial palsy. PMID- 11417616 TI - Magnetic resonance imaging findings in patients with severe neonatal indirect hyperbilirubinemia. AB - The aim of this study was to document the magnetic resonance imaging (MRI) findings of cases with a history of severe neonatal indirect hyperbilirubinemia. Ten cases (eight cases with neurologic findings, two normal cases) with a history of severe neonatal indirect hyperbilirubinemia were studied. Neurologic findings and MRI results were described and correlated. Seven of eight cases with neurologic findings demonstrated symmetric and uniform increased T2 signal changes limited to globus pallidi. MRI scans of two cases without neurologic findings showed no abnormality. Severe neonatal indirect hyperbilirubinemia should be considered in the differential diagnosis of bilateral symmetric hyperintense signal changes in the globus pallidus on MRI. However, high levels of unconjugated bilirubin concentrations in the neonatal period may not always cause such lesions of globus pallidus on MRI despite the presence of neurologic findings. PMID- 11417617 TI - Heliox use in the treatment of acute hyperammonemia. AB - The objective of this study was to evaluate a new method for the treatment of acute hyperammonemia with a helium-oxygen mixture (heliox). We conducted a prospective, randomized, controlled study of male Sprague-Dawley rats. Experimental hyperammonemia was induced by 7 days of a high-ammonia diet. Subsequently, the animals were randomly divided into two groups: the study group treated with heliox breathing for 24 hours and a control group breathing room air for 24 hours. A prospective, randomized, controlled laboratory animal study was conducted at an animal research facility. The baseline plasma ammonia level was 9.49 +/- 10.96 micromol/L. After 7 days of a high-ammonia diet, the plasma ammonia level rose to 31.53 +/- 8.86 micromol/L. There was a significant statistical difference between the plasma ammonia level following 24 hours of heliox therapy (23.14 +/- 13.97 micromol/L) and the ammonia level in the control group (42.31 +/- 24.25 micromol/L) (P < .05). Heliox breathing was found to be an efficient treatment modality for decreasing plasma ammonia levels in an animal model. Further studies are required to evaluate its potential application in the treatment of patients with hyperammonemia. PMID- 11417618 TI - Benign intracranial hypertension associated with budesonide treatment in children with Crohn's disease. AB - Oral budesonide in adult studies is a potent corticosteroid with decreased systemic bioavailability and an improved adverse effect profile in comparison with prednisone. It has recently been introduced for the treatment of inflammatory bowel disease in Europe, Canada, and Israel. Benign intracranial hypertension has rarely been associated with corticosteroid therapy but has not been reported in association with budesonide therapy. Three adolescents with Crohn's disease and poor nutritional status developed benign intracranial hypertension while receiving oral budesonide. All three patients had previously received multiple courses of prednisone during the course of their disease, without developing intracranial hypertension. Benign intracranial hypertension resolved after medication withdrawal and did not recur with subsequent use of prednisone. Evaluation for benign intracranial hypertension should be considered in patients with inflammatory bowel disease who develop headache while receiving oral budesonide. This side effect may be associated with poor nutritional status. PMID- 11417619 TI - Acute necrotizing encephalopathy presenting as a basal ganglia syndrome. AB - Acute necrotizing encephalopathy is a relatively new disease. The characteristic clinical findings are of febrile illness followed by rapid deterioration in mental status and seizures. The hallmark of the disease is multifocal bilateral symmetric lesions affecting the thalamus, hypothalamus, brainstem tegmentum, cerebral white matter, and cerebellum. The etiology is unknown, but immune mediated mechanism was suggested. We present a 12-year-old previously healthy girl who developed increased sleepiness progressing to stupor and coma. Magnetic resonance imaging (MRI) of the brain showed the characteristic findings previously described in acute necrotizing encephalopathy. Her mental status improved dramatically with steroid treatment, and the MRI findings resolved completely within 6 months. Following the acute illness, she developed a complex neuropsychiatric disorder consistent with basal ganglia syndrome. PMID- 11417620 TI - An eponymous experience. PMID- 11417623 TI - Determination of phytase activity in feed by a colorimetric enzymatic method: collaborative interlaboratory study. AB - Fourteen laboratories participated in a collaborative study (coded fyt9404) and 13 laboratories participated in a study (coded fyt9410) to validate a colorimetric assay for determination of microbial phytase activity in feed. For each study, all laboratories received 6 laboratory samples provided by one commercial supplier (phytase activity levels within the range of 200-400 per kg) to be analyzed in duplicate. Method performance was calculated and statistical calculations were executed according to AOAC guidelines. Results from 3 laboratories for study fyt9404 and from one laboratory for study fyt9410 were excluded from statistical analysis because of invalid data determined during initial review by Youden pair, value versus laboratory. For study fyt9404, repeatability relative standard deviation (RSDr) values ranged from 6.2 to 8.6%, and reproducibility relative standard deviation (RSDR) values ranged from 14.1 to 27.6%. No outliers were identified. For study fyt9410, RSDr values ranged from 3.9 to 7.9%, and RSDR values ranged from 14.0 to 20.5%. With outliers excluded, RSDr values ranged from 2.5 to 7.9%, and RSDR values ranged from 14.0 to 20.5%. PMID- 11417624 TI - Determination of DiazaCon in quail feed and quail serum by ion pair reversed phase chromatography. AB - Liquid chromatographic (LC) methods were developed for quantitating the potential avian contraceptive DiazaCon in quail feed and serum. DiazaCon was extracted from ground quail feed with basic n-butyl chloride. The n-butyl chloride extract was evaporated to dryness. The DiazaCon residues were dissolved in an aqueous methanolic ion pairing solution and quantitated by LC at 206 nm. Avian sera was combined with an equal volume of a pH 4 aqueous solution of ion pairing reagent and filtered to remove interfering proteins. DiazaCon was quantitated by LC. Mean recoveries for 500 and 2000 ppm fortified feed were 89.1 and 91.0%, respectively. The mean recovery for sera fortified at 5 levels ranging from 35 to 2000 ppm was 84.9%. Method limits of detection were approximately 14 and 13 ppm for feed and sera, respectively. PMID- 11417625 TI - LC/MS confirmation of ionophores in animal feeds. AB - A liquid chromatographic/mass spectrometric (LC/MS) electrospray confirmation method has been developed to confirm 4 ionophores (monensin, lasalocid, salinomycin, and narasin) in a variety of animal feeds using a single quadrupole mass spectrometer. The sodium ions of these compounds are dominant in the electrospray mass spectrum. Using optimized "in-source" collision induced dissociation, characteristic fragment ions seen previously using MS/MS can be observed. The drugs were extracted from the feed matrix using hexane-ethyl acetate and isolated using a silica solid-phase extraction cartridge. These ionophores were confirmed in both medicated feeds and nonmedicated feeds fortified with these drugs at the 1-50 ppm level. In addition, this method was used to confirm residues of monensin in a nonmedicated feed that was collected from a feed mill immediately after the production of a similar feed that was medicated with high levels of monensin. PMID- 11417626 TI - Voltammetric behavior and analytical applications of lomefloxacin, an antibacterial fluorquinolone. AB - Lomefloxacin was reduced on a dropping mercury electrode, producing one or more peaks, depending on the pH of the aqueous medium. Coulometric measurements gave an experimental value of 1 electron for the main peak. Electrolysis was followed by UV spectrophotometry and liquid chromatography (LC), showing that a new band at 413 nm appeared for the electrolysis product in an acidic medium. Furthermore, by using UV spectrophotometry, an apparent pKa value of 6.75 +/- 0.05 was obtained for lomefloxacin corresponding to the carboxyl moiety in the 3-position. For analytical studies, the differential pulse polarographic mode in 0.1 N HCl was selected. The repeatability and reproducibility of the method were adequate (coefficient of variation [CV], 0.51%). The calibration curve method was used for the lomefloxacin concentration range of 7.0 x 10(-6) to 7.0 x 10(-5)M. The detection and quantitation limits were 1.0 x 10(-6) and 6.9 x 10(-6)M, respectively. For purposes of comparison, both UV spectrophotometric and LC (with UV and fluorimetric detection) methods were developed. The polarographic method showed good selectivity with respect to both excipients and degradation products. The recovery study showed a CV of <2% and an average recovery of 99.5% and it was not necessary to treat the sample before analysis. The method was applied to the determination of the uniformity content of lomefloxacin commercial tablets. The polarographic method was also successfully applied to the quantitation of lomefloxacin in urine, and the renal excretion profile was also determined. PMID- 11417627 TI - Determination of flunixin residues in bovine muscle tissue by liquid chromatography with UV detection. AB - A new and sensitive liquid chromatography-ultra violet method with a detection limit of 6 ng/g (ppb) and a limit of quantification of 15 ng/g was developed for the determination of flunixin residues in bovine muscle tissue. Flunixin in homogenized animal tissue was extracted with acetonitrile after enzyme digestion. The tissue digest (extract) was then cleaned up on a solid-phase extraction cartridge and eluted with acidified hexane. After the eluate was evaporated to dryness under nitrogen at 55 degrees C, the residue was reconstituted in 1 mL mobile phase solution and analyzed by reversed-phase gradient chromatography with UV detection at 285 nm. The method was then applied in a survey study of slaughter animals to determine whether flunixin is being used in an off-label manner for veal and beef production in Canada. PMID- 11417628 TI - Occurrence of 1,4-dioxane in cosmetic raw materials and finished cosmetic products. AB - Surveys of cosmetic raw materials and finished products for the presence of the carcinogen 1,4-dioxane have been conducted by the U.S. Food and Drug Administration since 1979. Analytical methods are described for the determination of 1,4-dioxane in ethoxylated cosmetic raw materials and cosmetic finished products. 1,4-Dioxane was isolated by azeotropic atmospheric distillation and determined by gas chromatography using n-butanol as an internal standard. A solid phase extraction procedure based on a previously published method for the determination of 1,4-dioxane in cosmetic finished products was also used. 1,4 Dioxane was found in ethoxylated raw materials at levels up to 1410 ppm, and at levels up to 279 ppm in cosmetic finished products. Levels of 1,4-dioxane in excess of 85 ppm in children's shampoos indicate that continued monitoring of raw materials and finished products is warranted. PMID- 11417629 TI - Molecular diagnosis of microbial contamination in cosmetic and pharmaceutical products: a review. AB - Molecular methodologies such as adenosine triphosphate (ATP) bioluminescence and polymerase chain reaction (PCR)-based assays provide rapid quality control analysis of cosmetic and pharmaceutical finished products and raw materials. Using a single enrichment broth for bacteria, yeast, and mold, ATP bioluminescence detected microbial contamination within 27 h. Samples were automatically lysed to release microbial ATP and light production was quantitated using the Celsis Optocomp. However, to maintain the detection time to within 27 h, different enrichment broths were required for neutralization of antimicrobial ingredients in finished products and to provide specific nutrients for growth optimization. To perform the PCR reaction, bacterial DNA was extracted using a Tris-EDTA-Tween 20-proteinase K buffer at 35 degrees C while yeast and mold DNA were extracted using a Tris-EDTA-SDS buffer at 95 degrees C. Extracted microbial DNA was added to Ready-To-Go PCR beads and specific DNA primers. The primers were targeted to amplify specific regions within Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhimurium, Burkholderia cepacia, Candida albicans, and Aspergillus niger. Furthermore, conserved bacterial ribosomal DNA sequences have also been used for sterility testing of samples. The results from these studies indicate that both ATP bioluminescence and PCR assays provide rapid, reliable, and cost effective methods for quality evaluation. This will ultimately result in faster product release and production optimization. PMID- 11417630 TI - Application of liquid chromatography to the simultaneous determination of acetylsalicylic acid, caffeine, codeine, paracetamol, pyridoxine, and thiamine in pharmaceutical preparations. AB - This paper describes a rapid reversed-phase liquid chromatographic method, with UV detection, for the simultaneous determination of acetylsalicylic acid, caffeine, codeine, paracetamol, pyridoxine, and thiamine in pharmaceutical preparations. A reversed-phase C18 Nucleosil column is used. The mobile phase consists of 2 successive eluants: water (5 min) and acetonitrile-water (75 + 25, v/v; 9 min), both adjusted to pH 2.1 with phosphoric acid. Before determination acetylsalicylic acid is completely converted to salicylic acid by alkaline hydrolysis. Salicylic acid, caffeine, paracetamol, pyridoxine, and thiamine are all detected at 285 nm, whereas codeine is detected at 240 nm. Calibration curves were linear for salicylic acid, caffeine, paracetamol, and pyridoxine in the range of 50-500 mg/L, and for codeine and thiamine in the range of 50-1000 mg/L. The method was applied to the analysis of 13 fortified commercial pharmaceutical preparations. Recoveries ranged from 92.6 to 105.5%, with relative standard deviations of 1.1-5.8%. PMID- 11417631 TI - Selective separation and simultaneous determination of trace levels of five types of fluorinated quinolone drugs by thin-layer chromatography/fluorescence densitometry. AB - This paper reports the determination of trace levels of 5 types of fluorinated quinolone drugs, i.e., ciprofloxacin, norfloxacin, enoxacin, pefloxacin, and ofloxacin, by thin-layer chromatography (TLC)/fluorescence densitometry. The new analytical method uses 2-step TLC development, selective separation, and simultaneous determination of the 5 drugs. The method was also applied to the determination of recoveries of standards of the 5 drugs in plasma and urine samples. The results show that the method has a wide linear range, high repeatability, and good stability. PMID- 11417632 TI - Determination of coumarin in fragrance products by capillary gas chromatography with electron capture detection. AB - A gas chromatographic (GC) method is described for the determination of coumarin in fragrance products. Coumarin was tentatively identified by retention time and confirmed by GC/mass spectrometry. The amount of coumarin was determined by external standard. The method was validated by conducting recovery studies from fortified fragrance products at several concentrations. Recoveries of coumarin ranged from 99 to 110%, with a relative standard deviation of 3.24. The method was used to survey a variety of fragrance products purchased in the metropolitan Washington, DC area, for coumarin. Seventy one percent of the products were found to contain coumarin at concentrations ranging from 0.002 to 0.61%. PMID- 11417633 TI - Separation and quantitation of hazardous wastes from abrasive blast media. AB - A sample of glass bead abrasive blasting material (ABM) waste, received from Robins Air Force Base (Georgia), was examined to determine whether the waste could be rendered nonhazardous by separating paint contaminants from the ABM. The sample was analyzed with size distribution and toxicity characteristics leaching procedure. A Microtrac analyzer was used to measure the size of fine particles ( 325 Tyler mesh), and scanning electron microscopy analysis was performed to identify the nature of the contaminants in the ABM waste. Tests using froth flotation, magnetic separation, desliming, and acid washing were conducted to develop a process for removing the contaminants. A pilot plant test using the developed process rendered 82.1% or the ABM waste material nonhazardous. PMID- 11417634 TI - GC/ITMS measurement of carbonyls and multifunctional carbonyls in PM2.5 particles emitted from motor vehicles. AB - A method was developed and tested to identify and quantitate carbonyls and multifunctional carbonyls in fine particulate matter (PM2.5; <2.5 microm aerodynamic diameter). The method relies on ultrasonic extraction of particulate matter on filters at -8 degrees C; derivatization with O-(2,3,4,5,6 pentafluorobenzyl) hydroxylamine (PFBHA), and PFBHA along with bis (trimethylsilyl) trifluoroacetamide (BSTFA); and detection of the derivatives by gas chromatography/ion trap mass spectrometry. Ultrasonic extraction of model compounds from enriched particles was affected by solvent polarity (water > methylene chloride > toluene-isopropanol (2 + 1, v/v). Water provided the highest recovery for dihydroxy acetone, pyruvic acid, and hydroxy acetone, compared to methylene chloride, and toluene-isopropanol. Lowering the ultrasonication bath temperature from 0 degrees to -8 degrees C improved the recoveries of the less water soluble and more volatile species-methacrolein, methyl vinyl ketone, 2,3 butanedione, and tolualdehyde. The power of the method was demonstrated by identification and quantitation of carbonyls and multifunctional carbonyls in sample extracts of fine particulate matter (PM2.5) collected in the Caldecott tunnel, CA. The identities of crotonaldehyde, 2,3-butanedione, glyoxal, 9H fluoren-9-one, glycolaldehyde, glyoxylic acid, levulinic acid, and 3 hydroxybenzaldehyde were confirmed by comparing the relative retention time and mass spectra of the analyte in the sample extract with an authentic standard. Quantitation of crotonaldehyde, glyoxal, 2,3-butanedione, glyoxylic acid, and levulinic acid was accomplished. This is the first report of glyoxylic acid, levulinic acid, and 3-hydroxybenzaldheyde in PM2.5 particles sampled in a roadway tunnel. It is also the first report of a C10 carbonyl with the molecular formula of C10H16O2, a hydroxy carbonyl with the molecular formula of C17H21NO2, and a hydroxy or dihydroxy carbonyl with the molecular formula of C16H14O2 or C9H10O3. The high-molecular weight hydroxy carbonyls, which were found only in the heavy duty (diesel) bore, may be tracers of diesel emissions in air. PMID- 11417635 TI - Determination of trace levels of manganese in various biological and environmental samples by atomic absorption spectrometry after solid-liquid extraction and preconcentration with the ion pair formed by the nitroso-S anion and the tetradecyldimethylbenzylammonium cation. AB - Manganese is quantitatively retained by 2-nitroso-1-naphthol-4-sulfonic acid (nitroso-S) and tetradecyldimethylbenzylammonium (TDBA) chloride on microcrystalline naphthalene in the pH range 9.5-10.6 from large volumes of aqueous solutions of various samples. After filtration, the solid mass consisting of the manganese complex and naphthalene is dissolved in 5 mL dimethylformamide and the metal is determined by flame atomic absorption spectrometry. Alternatively, the manganese complex can be quantitatively adsorbed on TDBA naphthalene adsorbent packed in a column and determined similarly. About 0.2 microg manganese can be concentrated in a column from 400 mL aqueous sample with a concentration as low as 0.5 ng/mL. Eight replicate determinations of manganese at 0.8 microg/mL gave a mean absorbance of 0.156 for the final solution with a relative standard deviation of 1.4%. The sensitivity for 1% absorption was 23 ng/mL. The interference of a large number of anions and cations was studied, and the optimized conditions developed were used for trace determinations of manganese in various alloys, and in biological and environmental samples. PMID- 11417636 TI - Solid-phase extraction, separation, and visible spectrophotometric determination of trace amounts of iron in water samples. AB - A simple and reliable method is presented for the rapid extraction, separation, preconcentration, and determination of iron as its bathophenanthroline complex by the use of octadecylsilica membrane disks and spectrophotometry. We evaluted extraction efficiency, the influence of sample matrix, type and optimum amount of extractant, flow rates of sample solution and eluent, pH, amounts of bathophenanthroline and hydroxylamine hydrochloride, breakthrough volume, and limit of detection. We also studied the effects of various cationic interferences on percent recovery of iron. Complete elution of the complex from disks was obtained with a minimal amount of solvent. The limit of detection of the proposed method is 0.080 ppb. The method was applied to the recovery and determination of iron in natural waters. PMID- 11417637 TI - Reveal 8-Hour Test System for detection of Escherichia coli O157:H7 in raw ground beef, raw beef cubes, and iceberg lettuce rinse: collaborative study. AB - Five different food types were analyzed by the Reveal for E. coli O157:H7 8-Hour Test System (Reveal 8) and either the U.S. Food and Drug Administration's Bacteriological Analytical Manual (BAM) culture method or the U.S. Department of Agriculture Food Safety Inspection Service (FSIS) culture method for the presence of E. coli O157:H7. A total of 27 laboratories representing academia and private industry in the United States and Canada participated. Food types were inoculated with E. coli O157:H7 at 2 different levels: a high level where predominantly positive results were expected, and a low level where fractional recovery was anticipated. During this study, 1,110 samples and controls were analyzed by both the Reveal 8 and by BAM or FSIS by each of the collaborators (2,220 samples in total). For each set of samples, 740 were artificially inoculated with E. coli O157:H7, and 370 were uninoculated controls. The Reveal 8 detected 528 presumptive positives of which 487 were confirmed positive by the BAM culture method. In comparison, BAM and FSIS detected 489 of the 740 artificially contaminated samples as positive. In an additional in-house study performed only on chilled and frozen raw ground beef, 240 artificially inoculated samples were analyzed by both the Reveal 8 and by FSIS. The Reveal 8 detected and confirmed 104 samples as positive compared to 79 confirmed positive by FSIS. PMID- 11417638 TI - Comparison of the Reveal 20-hour method and the BAM culture method for the detection of Escherichia coli O157:H7 in selected foods and environmental swabs: collaborative study. AB - Four different food types along with environmental swabs were analyzed by the Reveal for E. coli O157:H7 test (Reveal) and the Bacteriological Analytical Manual (BAM) culture method for the presence of Escherichia coli O157:H7. Twenty seven laboratories representing academia and private industry in the United States and Canada participated. Sample types were inoculated with E. coli O157:H7 at 2 different levels. Of the 1,095 samples and controls analyzed and confirmed, 459 were positive and 557 were negative by both methods. No statistical differences (p <0.05) were observed between the Reveal and BAM methods. PMID- 11417639 TI - Detex for detection of Escherichia coli O157 in raw ground beef and raw ground poultry. AB - A method for detection of Escherichia coli O157 in beef and poultry is presented. The method is antibody-based and uses a patented antibody-specific metal-plating procedure for the detection of E. coli O157 in enriched meat samples. Both raw ground beef and raw ground poultry were tested as matrixes for the organism. The sensitivity and specificity of the assay were 98 and 90%, respectively. The accuracy of the assay was 96%. Overall, the method agreement between the E. coli O157 Detex assay and the U.S. Department of Agriculture/Food Safety Inspection Service method was 96%. Sample temperature upon loading of the apparatus was critical to the observed false-positive rate of the system. PMID- 11417640 TI - Determination of ephedrine-type alkaloids in dietary supplements by LC/MS using a stable-isotope labeled internal standard. PMID- 11417641 TI - Distribution among sample test results when testing shelled corn lots for fumonisin. AB - The statistical distribution known as the compound gamma function was studied for suitability in describing the distribution of sample test results associated with testing lots of shelled corn for fumonisin. Thirty-two 1.1 kg test samples were taken from each of 16 contaminated lots of shelled corn. An observed distribution consisted of 32 sample fumonisin test results for each lot. The mean fumonisin concentration, c, and the variance, s2, among the 32 sample fumonisin test results along with the parameters for the compound gamma function were determined for each of the 16 observed distributions. The 16 observed distributions of sample fumonisin test results were compared with the compound gamma function using the Power Divergence test. The null hypothesis that the observed distribution could have resulted from sampling a family of compound gamma distributions was not rejected at the 5% significance level for 15 of the 16 lots studied. Parameters of the compound gamma distribution were calculated from the 32-fumonisin sample test results using the method of moments. Using regression analysis, equations were developed that related the parameters of the compound gamma distribution to fumonisin concentration and the variance associated with a fumonisin test procedure. An operating characteristic curve was developed for a fumonisin sampling plan to demonstrate the use of the compound gamma function. PMID- 11417642 TI - Identification of Nile perch (Lates niloticus), grouper (Epinephelus guaza), and wreck fish (Polyprion americanus) fillets by PCR amplification of the 5S rDNA gene. AB - Nile perch (Lates niloticus), grouper (Epinephelus guaza), and wreck fish (Polyprion americanus) were differentiated by polymerase chain reaction (PCR) amplification of the nuclear 5S rDNA gene. The design of 3 species-specific primers complementary to the nontranscribed intergenic spacer region from the 5S rDNA molecule allowed amplification of clearly distinguishable gene fragments in each fish species. This approach is rapid and reliable and offers the potential to detect fraudulent or unintentional mislabeling of these species in routine seafood authentication analysis. PMID- 11417643 TI - Identification of allantoin, uric acid, and indoxyl sulfate as biochemical indicators of filth in food packaging by LC. AB - A liquid chromatographic (LC) method was developed for the determination of allantoin, uric acid, and indoxyl sulfate in mammalian urine contaminated packaging material including paper bagging, corrugated cardboard, grayboard, and burlap bagging. The procedure involves solvent extraction and isolation of the 3 analytes by reversed-phase LC with ultraviolet detection at 225 nm for allantoin and 286 nm for uric acid and indoxyl sulfate. The composition of authentic mammalian urine such as mouse, rat, cat, dog, and human were also determined with regard to the 3 compounds of interest. A linear concentration range of 0.11-20.4, 0.02-10.0, and 0.04-30.0 microg/mL was obtained for allantoin, uric acid, and indoxyl sulfate, respectively. Limits of detection (LOD) and quantitation (LOQ) were 0.0104 and 0.0345 microg/mL for allantoin; 0.0018 and 0.0060 microg/mL for uric acid; and 0.0049 and 0.0165 microg/mL for indoxyl sulfate, respectively. Interday relative standard deviation values for a mixture of standard allantoin, uric acid, and indoxyl sulfate (n = 5) were 0.97, 0.80, and 0.94%, respectively. Analyte composition for 5 types of authentic mammalian urine varied from 0.19 6.88 mg/mL allantoin; 0.08-0.57 mg/mL uric acid; and 0.03-0.78 mg/mL indoxyl sulfate. Analyte content for 8 samples including 2 samples each for paper, cardboard, grayboard, and burlap bagging each contaminated with mouse or rat urine ranged from 6.7%; confidence interval (CI) = +/- 1.4 microg/g; n = 27. Peer laboratory.--x = 56.6 +/- 6.6 microg/g; RSD = >11.7%; CI =+/- 4.1 microg/g; n = 8. PMID- 11417645 TI - Extension of AOAC Official Method 996.01 to the analysis of Standard Reference Material (SRM) 1846 and infant formulas. AB - There is currently no official method for the analysis of fatty acids (including trans fatty acids) in infant formulas. AOAC Official Method 996.01 for Fat Analysis in Cereal Products was extended to the analysis of milk-based infant formula Standard Reference Material (SRM)1846 to determine its applicability for use with infant formulas. Following the analysis of SRM 1846, 2 infant formulas, one milk-based liquid and one soy-based powdered infant formula, were analyzed for total fatty acid composition. Fatty acid methyl esters were prepared and analyzed by gas chromatography. The results of the analysis of SRM 1846 show that the mean analyzed values were highly reproducible as indicated by low coefficients of variation (CV). The CVs were <5% for the major fatty acids. Mean analyzed values for individual fatty acids in SRM 1846 were within +/- 1 standard deviation of the certificate values. The analyzed value for total fat as triglycerides (26.27 +/- 0.25%) agreed well with the certificate value (27.1 +/- 0.59%). Analyses of infant formulas showed that the concentrations of linoleic acid and fat meet the requirements for such formulas. PMID- 11417646 TI - Determination of phenolic disinfectant agents in commercial formulations by liquid chromatography. AB - Using liquid chromatography (LC), a convenient and versatile method was developed for the assay of disinfectant products containing the commonly used phenolic agents, alone or in combination, including phenol, 2-phenylphenol, 4-t amylphenol, 4-chloro-3,5-dimethylphenol (PCMX), 2-benzyl-4-chlorophenol, and triclosan. The procedure involves a simple dilution or dissolution of the product aliquot or portion followed by membrane filtration and LC. The method was applied to 19 different products representing 17 manufacturers and containing from one to 4 phenolic agents. Chromatography was performed using a Zorbax SB-C18 column, acetonitrile-0.05M KH2PO4 (55 + 45, v/v) pH 3.00, as mobile phase and UV detection at 280 nm. The intralaboratory precision of the product assays ranged form 0.34-5.35% (n = 5) and recoveries via fortification varied from 96.1-103.8%. The limits of quantitation (LOQ) and detection (LOD) ranged from 5.58 x 10(-5) to 2.50 x 10(-4) mg/mL and from 1.76 x 10(-5) to 0.67 x 10(-4) mg/mL, respectively, for the 6 analytes. The response for all analytes was observed to be linear for at least a 50-fold range in concentration (0.001-0.05 mg/mL). The proposed method provides an efficient means for the isolation and quantitation of these phenolic compounds. PMID- 11417647 TI - Determination by glyphosate and aminomethylphosphonic acid in crops by capillary gas chromatography with mass-selective detection: collaborative study. AB - A collaborative study was conducted to validate a method for the determination of glyphosate and aminomethylphosphonic acid (AMPA) in crops. The analytes are extracted from crops with water, and the crude extracts are then subjected to a cation exchange cleanup. The analytes are derivatized by the direct addition of the aqueous extract into a mixture of heptafluorobutanol and trifluoroacetic anhydride. The derivatized analytes are quantitated by capillary gas chromatography with mass-selective detection (MSD). The collaborative study involved 13 laboratories located in 5 countries; 12 laboratories returned valid data sets. The crops tested were field corn grain, soya forage, and walnut nutmeat at concentrations of 0.050, 0.40, and 2.0 mg/kg. The study used a split level pair replication scheme with blindly coded laboratory samples. Twelve materials were analyzed, including 1 control and 3 split-level pairs for each matrix, 1 pair at each nominal concentration. For glyphosate, the mean recovery was 91%, the average intralaboratory variance, the repeatability relative standard deviation (RSDr), was 11%, and the interlaboratory variance, the reproducibility relative standard deviation (RSDR), was 16%. For AMPA, the mean recovery was 87%, the RSDr was 16%, and the RSDR was 25% at mg/kg levels. PMID- 11417648 TI - Speciation of zinc in low molecular weight proteins of breast milk and infant formulas by size exclusion chromatography/flame atomic absorption spectroscopy. AB - Size exclusion chromatography (SEC) and flame atomic absorption spectroscopy (FAAS) were used for the separation of metal-containing species in milk whey. After milk ultracentrifugation, the sample was injected into a TSK-Gel G2000 glass column and eluted with 0.2M NH4NO3-NH3, pH 6.7. Low molecular weight proteins were fractionated, and the fractions were characterized by molecular weight. Zinc distributions were obtained by FAAS using a high performance nebulizer. The method was very sensitive (limit of detection = 2.6 x 10(-3) microg/mL; limit of quantitation = 8.9 x 10(-3) microg/mL) and precise (RSDs < or =10%). This method was applied to the determination of Zn in binding compounds in breast milk whey and in commercial cow's milk-based formulas. Distribution patterns were different. The presence of Zn in most fractions in breast milk was most significant, whereas in infant formulas Zn was detected only in fractions of molecular weight <5 kDa and in the highest molecular weight peak. PMID- 11417649 TI - Rapid determination of nineteen chlorophenols in wood, paper, cardboard, fruits, and fruit juices by gas chromatography/mass spectrometry. AB - Although the use of pentachlorophenol (PCP) is prohibited in most countries, it is still widely found in the wood of pallets, containers, crates and in cardboard, paper, etc. Such material may contain not only PCP but other chlorophenols as well. Wooden crates and cardboard boxes are often used to store and transport fresh fruits. Consequently, chlorophenols present in the wood may contaminate the stored fruits by migration. To ascertain that packaging, storage material, and fruit raw materials are free from chlorophenol residues, we developed a rapid and sensitive method for the detection of 19 chlorophenols by gas chromatography/mass spectrometry (GC/MS). The test portions are directly extracted and acetylated in a solution of sodium carbonate in the presence of acetic anhydride and hexane. The hexane layer is analyzed, without further purification, by GC/MS in the selective-ion monitoring mode. The method allows the analysis of >40 samples/day with detection limits of <20 microg/kg for chlorophenols in wood, cardboard, and paper, and <2 microg/kg for chlorophenols in fruits. Typical recoveries of all chlorophenols were 94% from paper, 115% from cardboard, 51% from wood (depending on the size of the chips or shavings), and 93% from fruit puree. PMID- 11417650 TI - Determination of lead and cadmium in ceramicware leach solutions by graphite furnace atomic absorption spectroscopy: method development and interlaboratory trial. AB - This method was developed to improve sensitivity and eliminate time consuming, evaporative pre-concentration in AOAC Method 973.82 and American Society for Testing and Materials method C738 for testing foodware. The method was developed using leach solutions obtained by leaching 9 differently decorated ceramic vessels with 4% acetic acid for 24 h at room temperature. Lead and cadmium concentrations in leach solutions were 0.005-17,600 and 0.0004-0.500 microg/mL, respectively. Concentrations were determined using peak area, phosphate chemical modifier (8.3 microg PO4(-3)), and a standard curve for quantitation. Optimized pre-atomization and atomization temperatures were 1,300 and 1,800 degrees C, respectively, for Pb and 1,100 and 1,700 degrees C, respectively, for Cd. Characteristic masses (mo) were 10 and 0.4 pg for Pb and Cd, respectively. Precision of repeated analyses of calibration solutions was < or =3% relative standard deviation. Precision of duplicate leach solution analyses on different days was 0-9% relative difference. Recovery from fortified leach solutions was 96 106%. Results obtained by this method agreed 92-110% with those of confirmatory analyses. Results of certified reference material solutions agreed 94-100% with certificate values. Pb and Cd limits of quantitation (LOQ) were 0.005 and 0.0005 microg/mL, respectively. Results from 3 trial laboratories for 4 leach solutions containing Pb and Cd concentrations of 0.017-1.47 and <0.0005-0.0864 microg/mL, respectively, agreed 89-102% with results of the author. Two attributes of this method were noteworthy: (1) Background absorbance due to organic matter was entirely absent from atomization profiles, making the use of short pre atomization hold times (2 s) possible. (2) Instrument precision was excellent and only one determination per solution was needed. Acceptance criteria for quality control measurements and a practical procedure for estimating the method LOQ during routine regulatory analyses are described. PMID- 11417651 TI - Gas chromatography with pulsed flame photometric detection multiresidue method for organophosphate pesticide and metabolite residues at the parts-per-billion level in representatives commodities of fruits and vegetable crop groups. AB - A gas chromatographic method with a pulsed flame photometric detector (P-FPD) is presented for the analysis of 28 parent organophosphate (OP) pesticides and their OP metabolites. A total of 57 organophosphates were analyzed in 10 representative fruit and vegetable crop groups. The method is based on a judicious selection of known procedures from FDA sources such as the Pesticide Analytical Manual and Laboratory Information Bulletins, combined in a manner to recover the OPs and their metabolite(s) at the part-per-billion (ppb) level. The method uses an acetone extraction with either miniaturized Hydromatrix column partitioning or alternately a miniaturized methylene dichloride liquid-liquid partitioning, followed by solid-phase extraction (SPE) cleanup with graphitized carbon black (GCB) and PSA cartridges. Determination of residues is by programmed temperature capillary column gas chromatography fitted with a P-FPD set in the phosphorus mode. The method is designed so that a set of samples can be prepared in 1 working day for overnight instrumental analysis. The recovery data indicates that a daily column-cutting procedure used in combination with the SPE extract cleanup effectively reduces matrix enhancement at the ppb level for many organophosphates. The OPs most susceptible to elevated recoveries around or greater than 150%, based on peak area calculations, were trichlorfon, phosmet, and the metabolites of dimethoate, fenamiphos, fenthion, and phorate. PMID- 11417652 TI - Determination of pesticides in composite dietary samples by gas chromatography/mass spectrometry in the selected ion monitoring mode by using a temperature-programmable large volume injector with preseparation column. AB - Use of a temperature-programmable preseparation column in the gas chromatographic (GC) injection port permits determination of a wide range of semi-volatile pesticides including organochlorines, organophosphates, triazines, and anilines in fatty composite dietary samples while reducing sample preparation time and solvent consumption. Dietary samples are mixed with diatomaceous earth and are Soxhlet-extracted with an azeotropic solution of hexane and acetone. Sample preparation uses liquid-liquid partitioning over diatomaceous earth followed by normal phase chromatography over partially deactivated alumina. The final cleanup step occurs in a preseparation column in the GC injector, which is able to perform splitless transfer of the analytes to the analytical column and purge 99% of the high molecular weight residue. Detection is performed by GC/mass spectrometry (MS) in the selected ion monitoring mode. Method detection limits were at or below 2 ng/g for 24 of 35 pesticides studied, with recovery between 70 and 125% for 27 pesticides in samples fortified at 10 ng/g. Recovery was not dependent on fat content when measured in laboratory fortified samples containing 1, 5, and 10% fat by weight. Precision over multiple injections was acceptable, with a relative standard deviation of 2.6-15% for 25 analytes. PMID- 11417653 TI - Determination of five pesticide residues in oranges by matrix solid-phase dispersion and liquid chromatography to estimate daily intake of consumers. AB - Residues of benzoylphenylurea insecticides (diflubenzuron, hexaflumuron, and flufenuxuron), carboxamide acaricides (hexythiazox), and carbamate insecticides (benfuracarb) were determined in 150 orange fruit samples from September 1998 to June 1999, to estimate exposure of the Valencian population to oranges contaminated with these newly developed pesticides. The method for monitoring these residues is based on matrix solid-phase dispersion and liquid chromatography with UV or atmospheric pressure chemical ionization/mass spectrometry (APCI/MS) detection. Orange samples representing 11 varieties were collected from an agricultural cooperative and examined for the 5 pesticides. In 74.6% of all analyzed samples, the pesticide residues were below detection limits, which ranged from 0.002 to 0.05 mg/kg. Residues were detected in 25.4% of the samples, with higher incidences of diflubenzuron, flufenuxuron, hexythiazox, and benfuracarb; hexaflumuron residues were detected only occasionally. Two different pesticides exceeded maximum residue limits (MRLs) in 4 (2.7%) of the orange samples. Diflubenzuron surpassed 1 mg/kg MRL in 3 samples and flufenuxuron exceeded the 0.3 mg/kg MRL in 3 samples. The estimated daily intake of the 5 pesticide residues during the period was 0.077 microg/kg body weight per day. This value is much lower than the total admissible daily intake proposed by the Food and Agricultural Organization and the World Health Organization. PMID- 11417654 TI - Simultaneous determination of residues of emamectin and its metabolites, and milbemectin, ivermectin, and abamectin in crops by liquid chromatography with fluorescence detection. AB - A liquid chromatographic (LC) method was developed for the determination of emamectin and its metabolites (8,9-Z-isomer, N-demethylated, N-formylated, and N methylformylated emamectin) in various crops. The analytes were extracted with acetone, cleaned up on cartridge columns (C18 and NH2), derivatized with trifluoroacetic anhydride and 1-methylimidazole, and determined by LC with fluorescence detection. Because radish inhibited the formation of the fluorescent derivatives, an additional Bond Elut PRS cartridge was used in the cleanup of Japanese radish samples. During sample preparation, N-formylated emamectin partially degraded to emamectin B1b and emamectin B1a, and the 8,9-Z-isomer partially degraded to N-demethylated emamectin. Therefore, emamectin and its metabolites were determined as total emamectin, i.e., their sum was estimated as emamectin benzoate. Their recoveries from most crops were approximately 80-110% with the developed method. The detection limits for the analytes in vegetables were 0.1-0.3 parts per trillion (ppt). The results for these compounds were confirmed by LC/mass spectrometry (LC/MS; electrospray ionization mode). Because the fluorescent derivative of emamectin was undetectable by LC/MS, the results for the analyte were confirmed by using a sample solution without derivatization. Limits of detection by LC/MS were similar to the fluorescence detection limits, 0.1-0.3 ppt in vegetables. In addition to the emamectins, milbemectin, ivermectin, and abamectin were also determined by the developed method. PMID- 11417655 TI - Examination of proficiency and control recovery data from analyses for pesticide residues in food: sources of variability. AB - We examined a number of large proficiency and control databases supporting the values reported for pesticide residues in agricultural commodities at fractions of a part per million (mg/kg). The average recovery from >100,000 recovery records in 13 databases was 94%. The overall average single-value relative standard deviation (RSD) of the reported recoveries was 17% at a mean concentration (C) of about 10(-7) (0.1 mg/kg). The average apparent HORRAT value (RSD found/RSDR predicted from the Horwitz formula [2*C(-0.1505)]) was 0.8. Analysis of variance indicated that about 60-70% of the variance could not be associated with any particular factor or combination of factors-analyte, commodity, method, laboratory, concentration, database, or their interactions. The most predominant factor, analyte, and its third-order interaction with laboratory and concentration contributed most of the assignable variance. These findings suggested that most of the variability of trace analysis for pesticide residues is "random" in the sense of being inherent and not assignable to specific factor fluctuations. PMID- 11417656 TI - Determination of heavy metals and pesticides in ginseng products. AB - Medicinal plants may carry residuals of environmentally persistent pesticides or assimilate heavy metals in varying degrees. Several factors may influence contaminant accumulation, including species, level and duration of contaminant exposure, and topography. As part of a program for assessment of the quality of herbal medicines, we have analyzed 21 over-the-counter ginseng (Panax ginseng) products in various dosage forms. Chromium, mercury, and arsenic were undetectable above their limits of detection in both liquid and solid samples; while cadmium, lead, and nickel were present in the majority of samples. The chlorinated pesticide levels varied widely. In most samples, the total concentration of pesticides was below 100 ppb; while in 5 samples the total concentration exceeded 100 ppb. PMID- 11417657 TI - Procedures for estimating confidence intervals for selected method performance parameters. AB - Procedures for estimating confidence intervals (CIs) for the repeatability variance (sigmar2), reproducibility variance (sigmaR2 = sigmaL2 + sigmar2), laboratory component (sigmaL2), and their corresponding standard deviations sigmar, sigmaR, and sigmaL, respectively, are presented. In addition, CIs for the ratio of the repeatability component to the reproducibility variance (sigmar2/sigmaR2) and the ratio of the laboratory component to the reproducibility variance (sigmaL2/sigmaR2) are also presented. PMID- 11417658 TI - A statistical model to evaluate analyte homogeneity for a material. AB - An underlying assumption for collaborative studies is that the analyte variation among test samples of the material (i.e., matrix and analyte concentration combination) under study has a negligible influence on the estimates of precision for the method. This assumption is expected to be fulfilled when the material under study is prepared (i.e., thoroughly mixed) such that the analyte is distributed uniformly throughout the matrix. Statistical design and intra-class correlation analysis procedures are proposed to assess the similarity or agreement among analytical results among- and within-containers for single and multiple occasions of use (e.g., collaborative and proficiency studies). PMID- 11417659 TI - 13C-IRIS: an improved method to detect the addition of low levels of C4-derived sugars to juices. AB - A method to improve the detection of cane or corn sugar addition to fruit juices has been developed. This involves the isolation of the individual sugars and measurement of their carbon isotope ratios. Using the Isotopic Relations of Individual Sugars approach, the additions of cane sucrose or cane invert as low as 4% of the total sugars can be detected in orange juice. This method offers a significant improvement over the conventional carbon isotopic method that has a detection limit of about 10%. PMID- 11417660 TI - Synergy between liquid chromatographic-pulsed amperometric detection and capillary-gas chromatographic methods for the detection of juice adulteration. AB - The use of liquid chromatography linked with pulsed amperometric detection, and capillary-gas chromatography to detect the adulteration of fruit juices is discussed. These are 2 complementary approaches which can be used to detect the addition of sugar syrups to juices at low levels. PMID- 11417661 TI - Isolation of lipid and 2-alkylcyclobutanones from irradiated foods by supercritical fluid extraction. AB - The 2-alkylcyclobutanone method was adopted as a European Standard (EN1785) and MAFF Validated Method (MAFF V37) in 1996 for the detection of irradiated food containing fat. As the method requires a relatively long period (ca 2 days) of time for extraction of the 2-alkylcyclobutanones from a foodstuff, a means was sought to increase the speed at which these irradiation markers could be isolated while at the same time decreasing the amount of organic solvents required. Thus, the technique of supercritical fluid extraction (SFE) was investigated. Results showed that SFE can be used for the rapid extraction (60 min) of lipid from irradiated foods such as chicken, pork, liquid whole egg, ground beef, and from the seeds of irradiated mango and papaya with only 10 mL n-hexane being necessary for collection of the extracted sample. A method was also developed whereby the 2 alkylcyclobutanones can be selectively extracted from irradiated foods without prior extraction of the lipid. The sample extract, in 10 mL n-hexane, is purified through a Florisil SPE cartridge which is washed with 10 mL n-hexane and the 2 alkylcyclobutanones eluted with 10 mL 2% diethyl ether in n-hexane before analysis by gas chromatography/mass spectrometry. 2-Dodecylcyclobutanone and 2 tetradecylcyclobutanone were selectively extracted from irradiated chicken meat, liquid whole egg, ground beef, and mango as well as from beef burgers and baked products containing irradiated ground beef and liquid whole egg, respectively. Using this method, samples can be analyzed for irradiation treatment within 6 h as opposed to the 2-day period required for the EN1785/MAFF V37 validated method. PMID- 11417662 TI - Routine analysis of ascorbic acid in citrus juice using capillary electrophoresis. AB - A procedure to monitor citrus juice samples was established to quantitate vitamin C by capillary electrophoresis using a previously developed method. Dilution and filtration were the only preparation requirements and separation was achieved with an uncoated capillary using a 35mM sodium borate buffer (pH 9.3) containing 5% (v/v) acetonitrile at 21 kV and 23 degrees C. Detection was performed by high speed scanning between 200 and 360 nm. From the multiwave length scan, the electropherogram at 270 nm was extracted and used to quantitate ascorbic acid. The ascorbic acid concentration was calculated with an internal standard method, with ferulic acid as internal standard. The level of ascorbic acid during analysis was stabilized with ethylenediaminetetraacetic acid and dithiothreitol was used to reduce dehydroascorbic acid to ascorbic acid to estimate the total vitamin C level. Results were similar to those obtained by liquid chromatography and the method is now used to determine routinely the level of ascorbic acid in citrus juices. PMID- 11417663 TI - Performance-Based instrument to assess functional capacity in dementia: The Texas Functional Living Scale. AB - OBJECTIVE: The Texas Functional Living Scale (TFLS) is a new performance-based measure of functional abilities with an emphasis on instrumental activities of daily living skills that is brief and weighted toward cognitive tasks. The purpose of this study was to examine its psychometric properties and clinical utility in patients with Alzheimer disease (AD) and normal elderly individuals. BACKGROUND: Measures of daily functional capacities used in patients with dementia often rely upon informant-based behavioral ratings or lengthy assessments of instrumental activities of daily living. Few existing quantitative tools emphasize performance-based cognitively-oriented tasks designed for patients with AD, although there is a need for such measures. METHOD: Twenty-two AD patients and 21 healthy controls were administered the TFLS and a measure of global cognitive status (Mini-Mental State Examination). Additionally, informant based ratings of daily living skills (Blessed Dementia Rating Scale) and emotional and behavioral status (Consortium to Establish a Registry for Alzheimer's Disease Behavior Rating Scale for Dementia) were obtained from caregivers of the AD patients. All subjects were re-evaluated approximately 1 month later using the same instruments. RESULTS: The TFLS showed a strong correlation with the Mini-Mental State Examination (r = 0.92). At 1 month, test retest reliability was high, and TFLS subscale scores were highly correlated (r > 0.90 for most) with the total score. The TFLS was moderately correlated with informant ratings of functional skills as assessed with the Blessed Dementia Rating Scale but was unrelated to emotional and behavioral symptoms as assessed with the Consortium to Establish a Registry for Alzheimer's Disease Behavior Rating Scale for Dementia. CONCLUSIONS: The TFLS showed evidence of good reliability, internal consistency, and convergent and discriminant validity with several popular measures of global cognitive status and behavioral functioning. It is a brief and easily administered performance-based measure of daily functional capabilities that is sensitive to level of cognitive impairment and seems applicable in patients with varying degrees of dementia. PMID- 11417664 TI - Motor instability in parkinsonian speech intensity. AB - OBJECTIVE: This study examined progressive speech intensity decay using two speech tasks: sustained vowel phonation (Experiment 1) and sentence reading (Experiment 2). BACKGROUND: Parkinsonian speech intensity has often been clinically observed to fade out or trail off. This gradual diminution of intensity is not unlike the well-documented progressive reduction of force underlying (upper limb) micrographic parkinsonian handwriting and (lower limb) marche a petit pas. Motor instability in speech intensity has yet to be investigated in a controlled experimental setting, however. METHOD: Thirteen Parkinson disease (PD) patients and their matched controls participated in Experiment 1: data from 6 PD patients and controls who naturally (i.e., without prior instruction) read the target sentence within a breath span were included in the analysis for Experiment 2. Participants were instructed to inhale maximally before vocalizing, and the extent of intensity declination over the breath span was measured. RESULTS: Parkinson disease patients demonstrated a consistently greater level of progressive intensity decay compared with matched controls for both speech tasks. This successful documentation and analysis of fading speech was interpreted as evidence for motor instability within the speech motor system in PD. CONCLUSIONS: It was concluded that the control of force in complex motor sequences involving speech and limb movement is affected by a common deficit in the frontostriatal circuit. PMID- 11417665 TI - Which symptoms of anxiety diminish after surgical interventions for Parkinson disease? AB - OBJECTIVE: The purpose of the current study was to address whether improvement in anxious symptoms after surgical treatment of Parkinson disease (PD) reflects a true reduction in anxiety as opposed to an epiphenomenon of parkinsonian symptom amelioration. BACKGROUND: Recent research suggests that anxiety is common in PD. An association between surgical intervention for PD and anxiety reduction has been reported; however, it is not clear which specific symptoms of anxiety improve. METHOD: Thirty-nine PD patients completed the Beck Anxiety Inventory (BAI) approximately 1 month before and 4 months after surgery. Twenty-four participants underwent unilateral pallidotomy, 10 underwent deep brain stimulating electrode implantation of the internal segment of the globus pallidus, 4 underwent thalamic deep brain stimulating electrode implantation, and 1 underwent left thalamotomy. RESULTS: Statistically significant reductions were found postoperatively in terms of BAI total score as well as neurophysiologic, autonomic, and subjective factors from the BAI. The panic factor did not significantly change after surgery, possibly secondary to limited power afforded by the sample size. CONCLUSIONS: Results suggest that surgical intervention for PD is associated with reduction in anxiety symptoms distinct from symptoms of PD. In other words, improvement in anxious symptoms reflects a true reduction in anxiety rather than simply being an epiphenomenon of parkinsonian symptom amelioration. PMID- 11417666 TI - Clinical and neuropsychological correlates of impaired awareness of deficits in Alzheimer disease and Parkinson disease: a comparative study. AB - OBJECTIVE: To compare patients with Alzheimer disease (AD) and Parkinson disease (PD) with regard to their awareness of cognitive, emotional/social interaction, self-care, and motor-related neurologic deficits. BACKGROUND: Unawareness of deficits, a clinically important symptom, is found in AD. It has been hypothesized to be associated with disruption of frontal-subcortical circuits but has been little studied in other neurodegenerative disorders. Because PD has a different anatomic-pathologic substrate, a comparison of impairment of awareness in AD and PD may shed light on the neural basis of this phenomenon. METHOD: Impairment of awareness was measured as the difference between patient self report and caregiver ratings of patient abilities on questionnaires tapping cognitive, emotional/social interaction, self-care, and motor function. These "discrepancy scores" were then compared between the two diagnostic groups and examined in relation to selected neuropsychological test data. RESULTS: In general, both AD and PD patients rate themselves as being less impaired than do their caregivers. The two diagnostic groups, AD and PD, differ significantly, however, on awareness discrepancy measures in the cognitive domain. In their ratings of patient cognitive skills, AD caregivers rate patients as significantly more impaired than patients rate themselves, whereas PD caregivers and patients do not differ significantly on these ratings. Impaired awareness in PD but not in AD is associated with poorer overall cognitive function and performance on tests measuring memory, attention, and constructional ability. CONCLUSIONS: Both AD and PD patients display impaired awareness of deficits in multiple domains, including motor-related neurologic function. Parkinson disease patients with comparatively intact cognitive function display relatively preserved awareness of motor and other deficits. PMID- 11417667 TI - Grip strength asymmetry in depressed boys. AB - OBJECTIVE: The purpose of this experiment was to investigate the influence of depressed mood on functional motor asymmetry among school-aged boys. Thus, left- and right-hand grip strength as well as fatigue across successive trials was examined among 38 right-handed school-aged boys, half of whom had been classified as depressed (n = 19) and the other half as nondepressed (n = 19). BACKGROUND: There is support in the literature that depressed individuals experience relative right anterior activation, and previous reports of functional motor asymmetries among depressed adults are generally supportive of this notion. To date, however, little is known regarding the neuropsychology of depression as it may pertain to children; in particular, the possibility of functional motor asymmetry within this population has yet to be explored. METHOD: Participants were asked to squeeze a hand dynamometer four times successively with each hand. The order of hand presentation was counterbalanced, and the dependent measure was in kilograms. RESULTS: As hypothesized, depressed boys failed to demonstrate asymmetry for grip strength, although nondepressed boys exhibited right-hand grip strength. In addition, based on trend, right-hand grip strength for depressed boys fatigued more rapidly across successive trials than did right-hand grip strength for nondepressed boys. In contrast, depressed boys maintained left-hand grip strength longer across successive trials than did nondepressed boys. CONCLUSIONS: The data are interpreted as consistent with current neuropsychological theories of emotion, particularly depression, and suggest that the functional motor asymmetries (i.e., failure to demonstrate asymmetric grip strength) observed in depressed adults are also present among depressed children. Implications for increased right hemisphere activation, decreased left hemisphere activation, or a combination of both are discussed. PMID- 11417668 TI - Foreign accent-like syndrome during psychotic exacerbations. AB - OBJECTIVE: The goal of this investigation was to describe the neurologic and psychiatric findings in a patient with foreign accent-like syndrome occurring during episodes of psychotic exacerbation. BACKGROUND: Foreign accent syndrome has been reported in several patients with disorders such as cerebral infarction, cerebral hemorrhage, and head trauma but not in a patient whose primary problem was psychosis. Most patients with this syndrome exhibit some degree of aphasia, and some are dysarthric. METHOD: A schizophrenic patient with foreign accent-like syndrome occurring during a psychotic exacerbation was evaluated by examination and interview, language testing, magnetic resonance imaging, electroencephalography, and other investigative methods. RESULTS: The patient exhibited a prominent British accent, which persisted throughout the duration of his psychotic exacerbation and resolved with improvement of his psychosis. Magnetic resonance imaging revealed no lesions, and no abnormality of language or articulation was present. A single-photon emission computed tomography scan could not be obtained. CONCLUSIONS: This represents the first reported case of a patient with foreign accent-like syndrome during psychotic exacerbations. PMID- 11417669 TI - Confessions of a drug user. PMID- 11417670 TI - Olfactory dysfunction in schizophrenia and temporal lobe epilepsy. AB - BACKGROUND: Schizophrenia and mesial temporal lobe epilepsy (TLE) represent two common brain disorders that share dysfunction of temporo-limbic neural substrates. OBJECTIVE: We evaluated whether patients with schizophrenia exhibited olfactory performance more similar to right or left temporal lobe epilepsy patients. METHODS: Odor identification ability and detection threshold sensitivity were measured in 40 patients with schizophrenia, 14 patients with right- and 18 patients with left-temporal lobe epilepsy (TLE) patients, and 25 healthy controls. Odor identification was assessed with the University of Pennsylvania Smell Identification Test (UPSIT) and odor detection threshold sensitivity with a single-staircase procedure using the stimulant phenyl ethyl alcohol (PEA). RESULTS: Relative to controls, only patients with schizophrenia and right TLE exhibited significant impairment in UPSIT performance. Left TLE patients and controls performed comparably on the UPSIT. Detection threshold sensitivity to PEA did not differ significantly among the four groups. CONCLUSIONS: These data suggest a greater reliance of olfactory processing on right hemisphere structures and are also consistent with recent neuroimaging studies that have implicated aberrant processing of olfactory information in right hemispheric brain regions in schizophrenia. PMID- 11417671 TI - Executive function and magnetic resonance imaging subcortical hyperintensities in vascular dementia. AB - OBJECTIVE: To determine the relation between subcortical hyperintensities (SHs) visible on magnetic resonance imaging and executive function among patients with vascular dementia. BACKGROUND: The relation between SHs and executive dysfunction is not well understood, because studies have varied widely in methodology and have produced conflicting results. METHOD: We examined the relation between SHs (expressed as a percentage of total brain volume, not including ventricular volume) and six tests of executive function in a well-defined group of 24 individuals with vascular dementia. Executive tests were divided in two groups: Attention/Speed and Abstraction/Problem Solving. Bivariate correlations were computed between individual neuropsychological variables and SHs. RESULTS: Results showed significant bivariate correlations between SHs and three of the four tests in the Attention/Speed domain. Subcortical hyperintensities shared virtually no association with performance on tests in the Abstraction/Problem Solving domain. CONCLUSIONS: The finding that SHs are significantly associated with psychomotor slowing and attentional dysfunction is consistent with what is known about the behavioral manifestations of subcortical disease. More detailed investigations of the regional distribution of SHs as well as measures of atrophy, hypoperfusion, and hypometabolism may be necessary to accurately characterize the complex relation between vascular disease and different aspects of executive dysfunction. PMID- 11417672 TI - Performance on the Hooper Visual Organizational Test in patients diagnosed with subcortical vascular dementia: relation to naming performance. AB - OBJECTIVE: To examine the performance of individuals with vascular dementia (VaD) on the Hooper Visual Organization Test (HVOT) and to determine the influence of naming on HVOT performance in this population. BACKGROUND: The HVOT is commonly administered to assess visuospatial perception among neurologic patients, but the test requires verbal identification of stimuli as well as spatial ability. Previous studies have examined the influence of naming on the HVOT, but the issue has not been systematically addressed in individuals with subcortical VaD. METHOD: Individuals with a diagnosis of VaD were administered the HVOT, three additional measures of visuospatial function (Block Design and Picture Completion subtests of the Wechsler Adult Intelligence Scale-Revised, Rey-Osterrieth Complex Figure Test copy trial), and the Boston Naming Test (BNT). RESULTS: On average, the VaD patients performed poorly compared with normative data on each of the cognitive measures, with the most pronounced deficit evident on the BNT. Regression analyses revealed that more than 60% of the variance in performance on the HVOT was accounted for by performance on the Block Design subtest of the Wechsler Adult Intelligence Scale-Revised; performance on the BNT did not contribute significantly. CONCLUSIONS: The results suggest that VaD patients perform below expectations on the HVOT and that the measure is robust to naming deficits in this population. PMID- 11417673 TI - Chronic cognitive disturbances after a single supratentorial lacunar infarct. AB - OBJECTIVE: At a minimum of 6 months after their stroke, 16 single supratentorial lacunar infarct (SSLI) patients and 16 controls underwent neuropsychological testing. After 6 months, neuropsychological testing was repeated to investigate the robustness and validity of the initial test results. BACKGROUND: An SSLI can cause subtle cognitive disturbances, which could explain the decrease in quality of life often observed in patients with these lesions. It is unclear at which stage after the occurrence of neurologic deficit possible neuropsychological impairment can be interpreted as chronic and at what moment neuropsychological testing can best be performed. METHOD: Patients and controls were administered a specifically developed neuropsychological test battery that included 14 standardized tasks tapping the main cognitive domains. Differences in test performances between the groups were analyzed by parametric statistics. RESULTS: At both examinations, similar abnormalities were found on five demanding neuropsychological tasks (language, concept shifting, abstraction, incidental memory, and verbal fluency), suggesting a generalized energetic deficiency. CONCLUSIONS: This long-term follow-up study showed evidence of subtle neurocognitive deficits after the occurrence of an SSLI, which are robust after 6 months. These mild cognitive disturbances may explain the decrease in quality of life that patients with an SSLI often experience. PMID- 11417674 TI - Diesel engines: environmental impact and control--a critical review introduction. PMID- 11417675 TI - Diesel engines: environmental impact and control. AB - The diesel engine is the most efficient prime mover commonly available today. Diesel engines move a large portion of the world's goods, power much of the world's equipment, and generate electricity more economically than any other device in their size range. But the diesel is one of the largest contributors to environmental pollution problems worldwide, and will remain so, with large increases expected in vehicle population and vehicle miles traveled (VMT) causing ever-increasing global emissions. Diesel emissions contribute to the development of cancer; cardiovascular and respiratory health effects; pollution of air, water, and soil; soiling; reductions in visibility; and global climate change. Where instituted, control programs have been effective in reducing diesel fleet emissions. Fuel changes, such as reduced sulfur and aromatics content, have resulted in immediate improvements across the entire diesel on- and off-road fleet, and promise more improvements with future control. In the United States, for example, 49-state (non-California) off-road diesel fuel sulfur content is 10 times higher than that of national on-road diesel fuel. Significantly reducing this sulfur content would reduce secondary particulate matter (PM) formation and allow the use of control technologies that have proven effective in the on-road arena. The use of essentially zero-sulfur fuels, such as natural gas, in heavy duty applications is also expected to continue. Technology changes, such as engine modifications, exhaust gas recirculation, and catalytic aftertreatment, take longer to fully implement, due to slow fleet turnover. However, they eventually result in significant emission reductions and will be continued on an ever-widening basis in the United States and worldwide. New technologies, such as hybrids and fuel cells, show significant promise in reducing emissions from sources currently dominated by diesel use. Lastly, the turnover of trucks and especially off-road equipment is slow; pollution control agencies need to address existing emissions with in-use programs, such as exhaust trap retrofits and smoke inspections. Such a program is underway in California. These and other steps that can be continued and improved will allow the use of the diesel engine, with its superior fuel consumption, to continue to benefit society while greatly reducing its negative environmental and health impacts. The next ten years can and must become the "Decade of Clean Diesel." PMID- 11417676 TI - The importance of pathogenic organisms in sewage and sewage sludge. AB - Deficient sanitation poses a serious threat to human and animal health, involving complex relationships between environments, animals, refuse, food, pathogens, parasites, and man. However, by sanitizing and stabilizing the organic matter of sewage sludge, agriculture can utilize it to maintain soil, water, and air quality. As ingredients in soil amendments, such bioresidues are a source of nutrients for plants. Stabilization and sanitation of sewage sludge safely couple its recycling and disposal. This coupling becomes increasingly important as economic and environmental constraints make strategies for waste disposal more difficult to apply. The occurrence of viruses, bacteria, yeasts, fungi, and zooparasites in sewage sludge is reviewed in this article, and consequential epidemiologic concerns that arise from sewage sludge recycling is also addressed. PMID- 11417677 TI - Application and field validation of a continuous nonmethane organic carbon analyzer. AB - Nonmethane organic carbon (NMOC) is a measure of total organic carbon except for that from CH4. We recently reported the development of online instrumentation for continuous NMOC monitoring. This instrument, referred to as C-NMOC, uses a microsorbent trap in combination with a gas-sampling valve as the sampling interface. A conventional oxidation/reduction NMOC detector is used for quantitation. In addition to being an online concentrator and an injector, the microtrap serves as a separator that isolates NMOC from H2O, CO, CO2, CH4, and other background gases. Therefore, the C-NMOC is able to handle high concentrations of background gases commonly found in stack emissions and has detection limits in the ppb levels. This paper reports the results of field validation and testing of a C-NMOC analyzer at a coatings facility in the eastern United States. The instrument was able to monitor the process transients in real time, based on which corrective actions could be taken. It demonstrated good accuracy, high precision, and long-term stability. PMID- 11417678 TI - A study of gas-phase mercury speciation using detailed chemical kinetics. AB - Mercury speciation in combustion-generated flue gas was modeled using a detailed chemical mechanism consisting of 60 reactions and 21 species. This speciation model accounts for the chlorination and oxidation of key flue-gas components, including elemental mercury (Hg0). Results indicated that the performance of the model is very sensitive to temperature. Starting with pure HCl, for lower reactor temperatures (less than approximately 630 degrees C), the model produced only trace amounts of atomic and molecular chlorine (Cl and Cl2), leading to a drastic underprediction of Hg chlorination compared with experimental data. For higher reactor temperatures, model predictions were in good accord with experimental data. For conditions that produce an excess of Cl and Cl2 relative to Hg, chlorination of Hg is determined by the competing influences of the initiation step, Hg + Cl = HgCl, and the Cl recombination reaction, 2Cl = Cl2. If the Cl recombination reaction is faster, Hg chlorination will eventually be dictated by the slower pathway Hg + Cl2 = HgCl2. PMID- 11417679 TI - A comparison of four gravimetric fine particle sampling methods. AB - A study was conducted to compare four gravimetric methods of measuring fine particle (PM2.5) concentrations in air: the BGI, Inc. PQ200 Federal Reference Method PM2.5 (FRM) sampler; the Harvard-Marple Impactor (HI); the BGI, Inc. GK2.05 KTL Respirable/Thoracic Cyclone (KTL); and the AirMetrics MiniVol (MiniVol). Pairs of FRM, HI, and KTL samplers and one MiniVol sampler were collocated and 24-hr integrated PM2.5 samples were collected on 21 days from January 6 through April 9, 2000. The mean and standard deviation of PM2.5 levels from the FRM samplers were 13.6 and 6.8 microg/m3, respectively. Significant systematic bias was found between mean concentrations from the FRM and the MiniVol (1.14 microg/m3, p = 0.0007), the HI and the MiniVol (0.85 microg/m3, p = 0.0048), and the KTL and the MiniVol (1.23 microg/m3, p = 0.0078) according to paired t test analyses. Linear regression on all pairwise combinations of the sampler types was used to evaluate measurements made by the samplers. None of the regression intercepts was significantly different from 0, and only two of the regression slopes were significantly different from 1, that for the FRM and the MiniVol [beta1 = 0.91, 95% CI (0.83-0.99)] and that for the KTL and the MiniVol [beta1 = 0.88, 95% CI (0.78-0.98)]. Regression R2 terms were 0.96 or greater between all pairs of samplers, and regression root mean square error terms (RMSE) were 1.65 microg/m3 or less. These results suggest that the MiniVol will underestimate measurements made by the FRM, the HI, and the KTL by an amount proportional to PM2.5 concentration. Nonetheless, these results indicate that all of the sampler types are comparable if approximately 10% variation on the mean levels and on individual measurement levels is considered acceptable and the actual concentration is within the range of this study (5-35 microg/m3). PMID- 11417680 TI - Combustion modeling and performance evaluation in a full-scale rotary kiln incinerator. AB - This work summarizes the results of numerical investigations and in situ measurements for turbulent combustion in a full-scale rotary kiln incinerator (RKI). The three-dimensional (3D) governing equations for mass, momentum, energy, and species, together with the kappa - epsilon turbulence model, are formulated and solved using a finite volume method. Volatile gases from solid waste were simulated by gaseous CH4 distributed nonuniformly along the kiln bed. The combustion process was considered to be a two-step stoichiometric reaction for primary air mixed with CH4 gas in the combustion chamber. The mixing-controlled eddy-dissipation model (EDM) was employed to predict the conversion rates of CH4, O2, CO2, and CO. The results of the prediction show that reverse flows occur near the entrance of the first combustion chamber (FCC) and the turning point at the entrance to the second combustion chamber (SCC). Temperature and species are nonuniform and are vertically stratified. Meanwhile, additional mixing in the SCC enhances postflame oxidation. A combustion efficiency of up to 99.96% can be achieved at approximately 150% excess air and 20-30% secondary air. Reasonable agreement is achieved between numerical predictions and in situ measurements. PMID- 11417681 TI - A fungal vapor-phase bioreactor for the removal of nitric oxide from waste gas streams. AB - Ground-level O3 formation is becoming a major concern in many cities due to recent tightening of O3 regulations. To control O3 formation, more efficient treatment processes for O3 precursors, such as NOx and volatile organic compounds (VOCs), are needed. One promising new technology for removing both NOx and VOCs from off-gas streams is biofiltration, a simple process whereby contaminated air is passed through a biologically active packed bed. In this study, a toluene degrading fungal bioreactor was used to treat an aerobic gas stream contaminated with NO. The fungal bioreactor removed 93% of the inlet 250-ppmv NO at an empty bed contact time (EBCT) of 1 min when supplied with 90 g/m3/hr toluene. The presence of NH4+ concentrations greater than 0.4 mg NH3/g dry packing medium, however, resulted in poor NO removal. The bioreactor achieved a maximum toluene elimination capacity of 270 g/m3/hr and maintained greater than 95% toluene removal efficiencies over the 175-day study period. PMID- 11417682 TI - Hydrolysis and heat treatment of aluminum dust. AB - Aluminum dust is a toxic and hazardous byproduct of Al remelting. The present research was performed to characterize and evaluate its behavior in water. The materials obtained by hydrolysis were also characterized, and the gases generated during the process were qualitatively analyzed. The effects of hydrolysis reaction time and temperature on the dust were also explored. The hydrolysis of Al dust is an exothermic reaction that gave rise to a solid composed of aluminum oxide, silicon oxide, and spinel (MgAl2O4). Most of the CH4, NH3, and SH2 gases generated were emitted immediately upon the start of the reaction, though their production continued for a long time. This slow reaction, which was moderately accelerated by temperature, led to the formation of a material less reactive than the untreated dust. On the other hand, heat treatment of the dust gave rise to an inert material composed of spinel, alumina, and magnesium and aluminum silicates. PMID- 11417683 TI - An outline of the 1994-1998 European inspection and maintenance study: part I design, tests, and results of experimental methods. AB - This paper provides an overview and the main results of a study aiming to evaluate current and alternative inspection and maintenance (I/M) programs to control emissions from in-use cars in Europe. For this purpose, a large number of passenger cars from five countries (France, Germany, Greece, the Netherlands, and the United Kingdom) were tested according to a common test protocol that included a variety of driving cycles and short tests, both legislative and specifically developed for this exercise. Evaluation of all test results was conducted with the objective of defining effective I/M test procedures and examining the potential of periodic I/M programs to reduce pollutant emissions and improve fuel economy of in-use cars. The results indicate that the current idle test used for catalyst-equipped cars in Europe is probably ineffective, while short transient loaded testing was found to offer higher potential in environmental terms. Regarding conventional gasoline cars, it is concluded that the idle test is sufficient, provided that the CO cut point is reduced and an HC test is added at high idle. Finally, the effectiveness of the free acceleration smoke test for diesels is questioned, while a transient cycle with continuous opacity measurement was found to be very promising. A companion paper further uses these results to analyze the likely cost-effectiveness on a per-country basis and discusses the possible effect of maintenance on emissions. PMID- 11417684 TI - Removal of hydrogen sulfide gas and landfill leachate treatment using coal bottom ash. AB - Coal bottom ashes produced from three thermal power plants were used in column and batch experiments to investigate the adsorption capacity of the coal ash. Hydrogen sulfide and leachates collected from three sanitary landfill sites were used as adsorbate gas and solutions, respectively. Experimental results showed that coal bottom ash could remove H2S from waste gas or reduce the concentrations of various pollutants in the leachate. Each gram of bottom ash could remove up to 10.5 mg of H2S. In treating the landfill leachate, increasing ash dosage increased the removal efficiency but decreased the adsorption amount per unit mass of ash. For these tested ashes, the removal efficiencies of chemical oxygen demand (COD), NH3-N, total Kjeldhal nitrogen (TKN), P, Fe3+, Mn2+, and Zn2+ were 36.4-50, 24.2-39.4, 27.0-31.1, 82.2-92.9, 93.8-96.5, 93.7-95.4, and 80.5-82.2%, respectively; the highest adsorption capacities for those parameters were 3.5 5.6, 0.22-0.63, 0.36-0.45, 0.027-0.034, 0.050-0.053, 0.029-0.032, and 0.006 mg/g of bottom ash, respectively. The adsorption of pollutants in the leachate conformed to Freundlich's adsorption model. PMID- 11417685 TI - Reproductive loss in high-producing dairy cattle: where will it end? AB - The dairy industry in the United States has changed dramatically in the last decade. Milk production per cow has increased steadily because of a combination of improved management, better nutrition, and intense genetic selection. Dairy farms are larger, and nearly 30% of the dairy cows in the United States are on farms with 500 or more cows. The shift toward more productive cows and larger herds is associated with a decrease in reproductive efficiency. Cows with the greatest milk production have the highest incidence of infertility, but epidemiological studies suggest that, in addition to milk production, other factors are probably decreasing reproductive efficiency in our dairy herds. The reproductive physiology of dairy cows has changed over the past 50 yr, and physiological adaptations to high milk production may explain part of the reproductive decline. Critical areas for new research include control of the estrous cycle, metabolic effects of lactation on reproduction, mechanisms linking disease to reproduction, and early embryonic mortality. Solving reproductive loss in dairy cows will not be easy because only a small number of research groups study reproduction in postpartum dairy cows. Therefore, the present research base will need to be expanded. For this to occur, research funding must be increased above its current level and a renewed emphasis must be placed on solving the emerging crisis of infertility in dairy cows. PMID- 11417686 TI - Invited review: adhesion mechanisms of rumen cellulolytic bacteria. AB - We divided the adhesion process of the predominant cellulolytic rumen bacteria Fibrobacter succinogenes, Ruminococcus flavefaciens, and Ruminococcus albus into four phases: 1) transport of the nonmotile bacteria to the substrate; 2) initial nonspecific adhesion of bacteria to unprotected sites of the substrate that is dominated by constitutive elements of bacterial glycocalyx; 3) specific adhesion via adhesins or ligands formation with the substrate, which can be dominated by several bacterial organelles including cellulosome complexes, fimbriae connections, glycosylated epitopes of cellulose-binding protein (CBP) or glycocalyx, and cellulose-binding domain (CBD) of enzymes; 4) proliferation of the attached bacteria on potentially digestible tissues of the substrate. Each of the phases and its significance in the adhesion process are described. Factors affecting bacterial adhesion are described including: 1) factors related to bacterial age, glycocalyx condition, and microbial competition; 2) factors related to the nature of substrate including, cuticle protection, surface area, hydration, and ionic charge; and 3) environmental factors including pH, temperature, and presence of cations and soluble carbohydrate. Based on the information available from the literature, it appears that each of the predominant rumen bacteria--F. succinogenes, R. flavefaciens, and R. albus--has a specific mechanism of adhesion to cellulose. In F. succinogenes, both the glycosidic residues of the outer membrane CBP and especially of the 180-kDa CBP, and the distinct CBD of EG2 EGF and Cl-stimulated cellobiosidase, may play a role in the adhesion to cellulose. No direct evidence, except scanning electron microscopy observations, yet supports the existence of either cellulosome complex or fimbriae structures involved in the adhesion mechanism of F. succinogenes. At least two mechanisms, cellulosome-like complexes and carbohydrate epitopes of the glycocalyx layer are involved in the specific adhesion of R. flavefaciens to cellulose. Ruminococcus albus possesses at least two mechanisms for specific adhesion to cellulose: a cellulosomal-like mechanism, and a CbpC (Pil)-protein mechanism that probably involves the production of fimbrial-like structures. Indirect and direct studies suggested that carbohydrate epitopes of CBPs and CBD epitope of cellulases may also be involved mostly in the nonspecific phase of adhesion of R. albus. PMID- 11417687 TI - Colloidal calcium phosphates in casein micelles studied by slow-speed-spinning 31P magic angle spinning solid-state nuclear magnetic resonance. AB - The composition of bovine casein micelles was analyzed by 31P magic angle spinning solid-state nuclear magnetic resonance spectroscopy. By looking at isotropic and anisotropic 31P chemical shift parameters, resonance line shapes, the combination of single-pulse and 1H to 31P cross-polarization spectra, and comparison with spectra for various model compounds combined with multiple component simulation and iterative fitting procedures, we were able to identify and quantify a variety of inorganic and organic phosphates in the micelles. These include phosphates from mobile and immobile inorganic hydroxyapatite-type phosphates as well as phosphates from kappa-casein and the Ca2+-binding phosphoserines from alphas1-, alphas2-, and beta-casein. This information is discussed in relation to previous knowledge and various models for the colloid formation. PMID- 11417688 TI - Effect of pasteurization of Ewe's milk and use of a native starter culture on the volatile components and sensory characteristics of roncal cheese. AB - The effect of pasteurization of milk and use of a native starter culture on the volatile components and sensory characteristics of a Spanish ewe's-milk cheese were examined. Three cheese batches were made, one from raw milk, another from pasteurized milk, and a third from pasteurized milk with an added native starter culture in addition to the commercial starter. Cheeses were analyzed at 1, 120, and 240 d of ripening. Analysis of the volatile components was by purge and trap connected to a gas chromatograph with a mass spectrometer and disclosed a total of 76 components belonging to the following chemical families: hydrocarbons, fatty acids, esters, sulfur and carbonyl compounds, and, in particular, alcohols. Pasteurization lowered the levels of certain volatile components, especially alcohols, aldehydes, and ketones. The cheeses made from pasteurized milk showed lower scores for attributes of characteristic taste and aftertaste, as well as a characteristic aroma at 240 d of ripening. These results suggest that the components present in higher concentrations in the cheeses made from the raw milk were necessary for development of characteristic Roncal cheese aroma. The new native starter culture tested did not exert a significant effect on any of the parameters considered, with the exception of certain isolated components, for which higher or lower quantities were recorded in the cheeses made with that starter culture, although the differences did not have a definite effect on the sensory characteristics of the cheeses. PMID- 11417689 TI - Short communication: influence of transglutaminase on the heat stability of milk. AB - Skim milk powders were prepared from control and transglutaminase-treated skimmed milk. The heat stability of reconstituted transglutaminase-treated skimmed milk (9.0% total solids) was markedly increased in the pH region of minimum stability (pH 6.8 to 7.1) compared with control milk, while the heat stability of reconstituted concentrated transglutaminase-treated skimmed milk (22.5% total solids) increased progressively as a function of pH relative to control milk. The effect of transglutaminase treatment on the heat stability of skimmed milk may have commercial applications, but extensive research is necessary to gain a better understanding of the mechanism by which transglutaminase improves heat stability. PMID- 11417690 TI - Influence of milk-clotting enzyme concentration on the alphas1-casein hydrolysis during soft cheeses ripening. AB - We studied the influence of the dose of milk-clotting enzyme on alphas1-CN degradation, soluble nitrogen production, and sensory profile for an Argentinean soft cheese: Cremoso Argentino. Five different types of cheeses were produced: 1) control cheeses with normal technology, 2) cheeses with inactivated milk-clotting enzyme, 3) cheeses with inactivated milk-clotting enzyme, without starter (acidified with glucono delta lactone), 4) cheeses with a half dose of milk clotting enzyme, and 5) cheeses with a double dose of milk-clotting enzyme. Proteolysis was assessed by isoelectric focusing electrophoresis of the insoluble fraction at pH 4.6, followed by densitometric quantification. Soluble nitrogen at pH 4.6, expressed as a percentage of total nitrogen and defined as ripening index was also performed. A sensorial panel evaluated the cheeses at the end of ripening. The hydrolysis level of alphas1-CN depended on the milk-clotting enzyme dose used in cheese making. Cheeses without active coagulant did not show degradation at the end of ripening, while cheeses with half and whole doses showed proportional degradations to coagulant dose. Cheese with a double dose of coagulant did not show higher alphas1-CN hydrolysis than normal cheese. No difference was found between cheeses with and without microbiological starter, indicating that the selected culture, composed of thermophilic strains, was unable to attack the whole casein. A high linear correlation was found between ripening index and the relation Sensorial characteristics of cheeses agree with objective analysis. Cheeses without active coagulant were hard and crumbly, while cheeses with normal dose were soft and creamy. PMID- 11417691 TI - Effectiveness of poly(ethylene terephthalate) and high-density polyethylene in protection of milk flavor. AB - The development of certain off-flavors in whole milk (3.25% milk fat) as related to packaging material [glass, high-density polyethylene (HDPE), amber poly(ethylene terephthalate) (PETE), clear PETE, and clear PETE-UV] were evaluated after exposure to fluorescent light (1100 to 1300 lx) for 18 d at 4 degrees C. Control samples packaged and stored under identical conditions were wrapped in foil to prevent light exposure. Selected flavor compounds in milk were measured analytically on d 0, 7, 14, and 18 of storage, while intensities of "oxidation," "acetaldehyde," and "lacks freshness" off-flavors were determined by sensory analysis at the same intervals. In light-exposed samples, oxidation off flavor was significantly lower when packaged in amber PETE versus other containers. Milk packaged in HDPE containers showed a significantly higher level of oxidation off-flavor than milk packaged in PETE-UV containers but not higher than clear PETE or glass containers. No significant difference in acetaldehyde off-flavor was found between package material treatments (exposed or protected). Acetaldehyde concentration never exceeded flavor threshold levels, regardless of packaging material. Amber and PETE-UV materials proved to be a competitive packaging choice for milk in preserving fresh milk flavor. PMID- 11417692 TI - Effect of milk preacidification on low fat mozzarella cheese: II. Chemical and functional properties during storage. AB - The effect of milk preacidification on cheese manufacturing, chemical properties, and functional properties of low fat Mozzarella cheese was determined. Four vats of cheese were made in 1 d using no preacidification (control), preacidification to pH 6.0 and pH 5.8 with acetic acid, and preacidification to pH 5.8 with citric acid. This process was replicated four times. Modifications in the typical Mozzarella manufacturing procedures were necessary to accommodate milk preacidification. The chemical composition of the cheeses was similar among the treatments, except the calcium content and calcium as a percentage of protein were lower in the preacidified treatments. During refrigerated storage, the chemical and functional properties of low fat Mozzarella were affected the most by milk preacidification to pH 5.8 with citric acid. The amount of expressible serum, unmelted cheese whiteness, initial unmelted hardness, and initial apparent viscosity were lower with preacidification. The reduction in initial unmelted cheese hardness and initial apparent viscosity in the pH 5.8 citric treatments represents an improvement in the quality of low fat Mozzarella cheese that allows the cheese to have better pizza bake characteristics with shorter time of refrigerated storage. PMID- 11417693 TI - Effect of milk preacidification on low fat mozzarella cheese: III. Post-melt chewiness and whiteness. AB - The effect of calcium reduction (as a result of milk preacidification) on post melt chewiness and whiteness of low fat Mozzarella cheese was determined. Four vats (230 kg of milk per vat) of cheese were made in 1 d using no preacidification (control), preacidification pH 6.0 and pH 5.8 with acetic acid, and preacidification to pH 5.8 with citric acid. Cheese manufacture was repeated on four different days using a randomized complete block design. The total calcium content and the water-insoluble calcium content of the cheese were lower in the cheeses made from preacidified milks. The amount of water-soluble and water-insoluble calcium changed during refrigerated storage, as did pH. The post melt chewiness and whiteness of low fat Mozzarella cheese were affected by milk preacidification. The largest level of calcium reduction and modification in post melt chewiness and whiteness occurred in the pH 5.8 citric treatment. Multiple regression analysis of post-melt chewiness and cheese whiteness at 38 degrees C after heating and cooling indicated that both water-insoluble calcium and proteolysis were strongly associated with changes in the post-melt chewiness and whiteness of low fat Mozzarella cheese. High levels of proteolysis and low levels of water-insoluble calcium were associated with decreased post-melt chewiness and whiteness of low fat Mozzarella cheese. PMID- 11417694 TI - Natural exopolysaccharides enhance survival of lactic acid bacteria in frozen dairy desserts. AB - Viable lactic acid-producing bacteria in frozen dairy desserts can be a source of beta-galactosidase for persons who absorb lactose insufficiently. However, freezing kills many of the cells, causing loss of enzymatic activity. Cultures selected for high beta-galactosidase activities and high survival rates in the presence of bile were examined for survivability during freezing in reduced-fat ice cream. Encapsulated S. thermophilus strains survived better than their nonencapsulated mutants in reduced-fat ice cream after freezing and frozen storage at -29 degrees C for 16 d (28 vs. 19%). However, a small nonencapsulated strain of Lactobacillus delbrueckii sp. bulgaricus survived better than the large encapsulated strain in reduced-fat ice cream. Factors that improved survival of encapsulated S. thermophilus 1068 in ice cream were 1) harvest of cells in the late-log phase of growth at 37 degrees C rather than at 40, 42.5, or 45 degrees C; 2) overrun at 50% rather than 100%; and 3) storage at -17 degrees C rather than -23 or -29 degrees C. Survival of strain ST1068 was unaffected by 1) neutralization of acid during growth or 2) substitution of nitrogen for air in building overrun. PMID- 11417695 TI - Milk fatty acid composition and mammary lipid metabolism in Holstein cows fed protected or unprotected canola seeds. AB - Six midlactation Holstein cows were fed a total mixed ration supplemented with either 4.8% canola meal, 3.3% unprotected canola seeds plus 1.5% canola meal, or 4.8% formaldehyde-protected canola seeds, according to a double 3 x 3 Latin square design. Each period lasted 3 wk; experimental analyses were restricted to the last week of each period. Mammary biopsies were taken the last day of each period for gene expression measurements. Milk production and milk protein percentage were reduced by canola seeds, whether protected or unprotected. Protected canola seeds also decreased dry matter intake. Feeding canola seeds reduced the content of C8 to C16 fatty acids in milk and increased the content of oleic acid (C18:1c9). Unprotected canola seeds elevated the concentrations of C18:0. Protected canola seeds increased the C18:2 and C18:3 content, and reduced the C18d:0/C18:1c9 ratio. Similar results were obtained for plasma fatty acids, with some specific features, such as an increased C16:0/C16:1 ratio with protected canola seeds. Canola seeds had no significant effects on insulin, triglycerides, or cholesterol present in serum, but increased the concentration of nonesterified fatty acids; a greater increase was obtained with protected canola seeds. Expression levels of acetyl-CoA carboxylase and delta 9-stearoyl CoA desaturase genes measured in the mammary gland did not differ significantly between diets. Therefore, the reduced C18s:0/C18:1c9 ratio observed in milk with protected canola seeds was not due to an enhanced expression of the delta-9 desaturase in the mammary gland. PMID- 11417696 TI - Sympathetic nervous system activity in adipose tissues during pregnancy and lactation of the rat. AB - The concentration and turnover of norepinephrine in white adipose and liver tissues were determined in pregnant, lactating, and age-matched virgin rats to elucidate the adaptations in sympathetic nervous system activity. In study 1, at d 18 of pregnancy and d 7 and 21 of lactation, animals were killed, and liver and cardiac perimetrial and retroperitoneal adipose depots were quick-frozen and then assayed for norepinephrine as a gross estimate of sympathetic innervation. In study 2, the same design was used to measure the turnover of norepinephrine as a measure of sympathetic activity. Animals were treated with alpha methylparatyrosine, an inhibitor of norepinephrine synthesis, and killed at 0, 1.5, and 3 h after injection. In pregnant animals, basal norepinephrine concentrations were decreased compared with unbred controls in perimetrial and retroperitoneal depots. By d 21 of lactation, all adipose depots from lactating animals had more norepinephrine than did controls. The turnover of norepinephrine decreased in noncardiac adipose depots of pregnant animals. By d 21 of lactation, norepinephrine concentration was greater in all of the adipose depots than in controls. The turnover rate was faster in all adipose tissue depots but only significantly different in the cardiac depot. Sympathetic nervous activity in adipose tissue is diminished in pregnant rats, presumably to save energy for fetal growth and maternal fat storage. In late lactation, activity is increased, presumably to direct fatty acids away from adipose tissue towards milk production. The data from this study are consistent with the hypothesis that the sympathetic nervous system plays a role in the regulation of adipose metabolism in lactation. PMID- 11417697 TI - Body condition related to ketosis and reproductive performance in Norwegian dairy cows. AB - The relationships between body condition score (BCS) and ketosis, and between BCS and reproductive performance in 732 moderate yielding, dual-purpose cows were studied. The cows were of the breed Norwegian Cattle. Farms with tie-stall barns and a history of high ketosis incidence were chosen for the study. Sixteen assessors visited the farms monthly and the same assessor assigned a BCS to each cow once a month. A BCS of 3.5 or higher at calving was associated with increased risk for ketosis. Cows that subsequently developed ketosis had higher BCS than healthy cows before the disease was diagnosed, and they lost more body condition than did the latter after ketosis had occurred. Summer calving cows and primiparous cows showed the lowest risk of ketosis. A history of ketosis before first service decreased the likelihood of conception to that service. Loss in body condition during the postpartum period was associated with decreased likelihood of conception to first service, prolonged calving to conception intervals and increased number of artificial inseminations per conception. Reproductive performance was not associated with BCS at calving. We concluded that BCS is a useful method of monitoring relations among nutritional management, reproduction, and ketosis in moderate yielding dual-purpose cows. PMID- 11417698 TI - Milk urea testing as a tool to monitor reproductive performance in Ontario dairy herds. AB - Dairy herd improvement test-day data, including milk urea concentrations measured using infrared test method, were collected from 60 commercial Ontario Holstein dairy herds for a 13-mo period between December 1, 1995, and December 31, 1996. The objective of the study was to describe, at the cow and the group level, the relationship between DHI milk urea concentrations and reproductive performance in commercial dairy herds. When interpreted at the cow level, there was no association between milk urea and the risk for pregnancy from an insemination occurring within the 45-d period preceding test day. However, a negative curvilinear relationship existed between milk urea and the risk for pregnancy from a first, second, or third insemination event occurring within the 45-d period following test day, with the odds for pregnancy being highest when the milk urea on the test day preceding the insemination was either below 4.5 mmol/L or greater than 6.49 mmol/L, compared with a concentration between 4.5 and 6.49 mmol/L. When interpreted at the group level, there was no association between group mean milk urea for cows between 50 and 180 DIM, and the group conception rate for cows receiving a first, second, or third insemination event in the 45-d period either preceding or following test day. Thus, while DHI milk urea measurements may be useful as a management tool to improve the efficiency of production or reduce nitrogen excretion, through helping to optimize the efficiency of protein utilization, they may have limited utility as a monitoring or diagnostic tool for reproductive performance. The results of this study suggest that good fertility may be achieved across a broad range of milk urea concentrations. PMID- 11417699 TI - Influence of subclinical mastitis during early lactation on reproductive parameters. AB - Our objective was to determine the effects of mastitis during early lactation on the reproductive performance of Jersey cows. From 1986 to 1997, quarter foremilk samples were collected every 4 to 8 wk during lactation, at drying off, near calving, and when clinical mastitis was diagnosed and were evaluated microbiologically to identify causative bacteria. Services per conception, days open, and days to first service were obtained from DHIA records on 752 cows. Cows were separated by mastitis type (clinical, n = 186; subclinical, n = 240; control, uninfected or infected after confirmed pregnancy, n = 326). Cows were reclassified based on the time of clinical or subclinical mastitis as follows: period 1, before first service (n = 374); period 2, between first service and pregnancy (n = 52); and period 3, after confirmed pregnancy or uninfected (control; n = 326). Milk production did not differ for any group separations. Reproductive performance did not differ between gram-negative or gram-positive mastitis pathogens. Cows with clinical or subclinical mastitis before first service had increased days to first service (77.3+/-2.7 and 74.8+/-2.7 d), days open (110.0+/-6.9 and 107.7+/-6.9 d), and services per conception (2.1+/-0.2 and 2.1+/-0.2) compared with controls (67.8+/-2.2 d, 85.4+/-5.8 d, 1.6+/-0.2; P < 0.05). Days to first service were not increased in cows with clinical or subclinical mastitis during period 2 (70.6+/-3.3 and 61.2+/-7.8 d). However, days open (143.6+/-8.5 d) and services per conception (3.0+/-0.2) were increased (P < 0.05) in cows with clinical mastitis during period 2, but not in cows with subclinical mastitis (90.9+/-20.2 d and 2.1+/-0.5). Cows initially diagnosed subclinical that became clinical during period 2 exhibited increased days to first service (93.9+/-10.1 d), days open (196.0+/-26.2 d), and services per conception (4.3+/-0.7) compared with control animals (P < 0.05). In conclusion, subclinical mastitis reduced reproductive performance of lactating cows similar to clinical mastitis. Subclinical mastitis followed by clinical mastitis resulted in the most severe loss in reproductive performance. PMID- 11417700 TI - Application of differential inflammatory cell count as a tool to monitor udder health. AB - A flow cytometric technique called differential inflammatory cell count was standardized by staining bovine peripheral blood leukocytes with a combination of DNA binding dyes SYBR green 1 and propidium iodide in water. Leukocytes were also stained with propidium iodide in detergent to determine total cell count. Differential inflammatory cell count assay was evaluated with individual quarter milk samples from 13 cows. Cows were sampled at weekly intervals for 3 wk and assayed for total cell count, mononuclear leukocyte count, and polymorphonuclear leukocyte count. Simultaneously, milk samples were evaluated by the conventional electronic somatic cell count (SCC) technique. Somatic cell count positively correlated with total cell count (r = 0.9), mononuclear leukocyte count (r = 0.8), and polymorphonuclear leukocyte count (r = 0.89). Quarters with SCC > log10 5.4 had a higher total cell count, mononuclear leukocyte count, and polymorphonuclear leukocyte count and were more often culture positive compared with quarters with SCC < log10 5.4. Quarters that were culture positive on all three test occasions had a higher proportion of polymorphonuclear leukocytes (33 to 49%) compared with quarters that were culture negative on all three test occasions (17 to 25%). The findings of this study suggest that differential inflammatory cell count assay has the potential to evolve as a new technique for evaluation of udder health status. PMID- 11417701 TI - Phenotypic and genotypic characterization of Pseudomonas fluorescens isolated from bulk tank milk. AB - Pseudomonas fluorescens isolates (n = 55) isolated from farm bulk tank milk (n = 55) from dairy herds in eastern South Dakota and western Minnesota were examined for phenotypic (biotype, proteolytic, and lipolytic profiles) and genotypic (plasmid profiles and 16S-23S PCR ribotypes) characteristics. The observed phenotypic and genotypic characteristics were used to conduct phylogenetic analysis. Pseudomonas fluorescens belonged to 28 API 20 NE biotypes and 14 proteolytic and lipolytic profiles. It was observed that 80, 91, and 58% of the isolates were proteolytic at 7, 22, and 32 degrees C, respectively. Only 7, 44, and 7% of the isolates were lipolytic at the same three temperatures. Pseudomonas fluorescens was more likely to produce proteinases at 7 and 22 degrees C and lipases at 22 degrees C. Only 9 of 55 isolates of P. fluorescens harbored plasmids. This small percentage of plasmid-bearing isolates provided insufficient data for inferences related to the distribution of plasmid-bearing clonal types. Based on 16S-23S PCR ribotyping, P. fluorescens belonged to 14 subtypes. The 16S 23S PCR ribotyping technique allowed differentiation between strains; however, it did not concur with the biotypes and proteolytic and lipolytic profiles. Use of biotypes in conjunction with proteolytic and lipolytic profiles might have practical value for conducting trace-back studies related to P. fluorescens. Based on phylogenetic analysis, it was inferred that for the given geographical area and time period, P. fluorescens isolated from farm bulk tank milk consists of a large heterogeneous group of organisms. PMID- 11417702 TI - Induction of nitric oxide production by bovine mammary epithelial cells and blood leukocytes. AB - A recent study from our laboratory has shown that significant amounts of nitric oxide are released by somatic cells recovered during endotoxin-induced mastitis. The present study was undertaken to investigate which cell type(s) among milk somatic cell population can produce nitric oxide under inflammatory conditions. Nitric oxide release from mammary epithelial cell lines and from bovine neutrophils and monocytes extracted from blood was measured in response to cytokines and Escherichia coli lipopolysaccharides. An epithelial cell line isolated from bovine mammary gland, FbE cells, was found to release nitric oxide after exposure to interleukin-1beta. This nitric oxide production was completely abolished by addition of L-N6-(1-iminoethyl) lysine, a potent inducible nitric oxide synthase inhibitor. Bovine monocytes produced nitric oxide in response to recombinant bovine interferon-gamma alone or in combination with E. coli lipopolysaccharides. In these cells, nitric oxide release was reduced by the addition of inducible nitric oxide synthase inhibitors L-N6-(1-iminoethyl) lysine and aminoguanidine. Lipopolysaccharides and recombinant bovine interferon-gamma increased nitric oxide synthase mRNA in neutrophils, but nitric oxide release could not be detected under any of the experimental conditions used. These results show that bovine epithelial cells and mononuclear phagocytes produce nitric oxide under inflammatory conditions and suggest that these cell populations are responsible for nitric oxide release observed during mastitis. PMID- 11417703 TI - Assessment of rumen processes by selected-ion-flow-tube mass spectrometric analysis of rumen gases. AB - This work investigated the potential to use measurement of the concentration of certain gases in the rumen headspace to gain information about rumen processes and as a potential diagnostic tool. We used new equipment (selected-ion-flow-tube mass spectrometer) that allows rapid and precise analysis of many of the gases present in a sample. Samples of rumen headspace gas and corresponding samples of rumen liquor were taken from three lactating cows, prepared with rumen fistulae, at intervals after receiving their morning feed allocation (grass silage and concentrates). Hydrogen sulfide, methyl sulfide, and dimethyl sulfide, were the predominant gases that were measured in the rumen headspace by this technique. The concentrations of these sulfur compounds declined over the interval after feeding, mirroring ammonia concentrations measured in rumen liquor, reflecting their common dependence on the fermentation of sulfur amino acids. Ammonia concentrations in rumen headspace gas varied in the opposite direction to the concentration of ammonia in rumen liquor and likely depend more on the pH of rumen liquor. Consideration of the pKa of ammonia suggests that ammonia concentrations in rumen gas will be very low below pH 6, representing a useful diagnostic for subacute ruminal acidosis. Low concentrations of volatile fatty acids were detected in rumen gas. The molar proportions of volatile fatty acids were similar in gas and liquor samples, with rumen gas containing slightly less acetic acid and disproportionately more valeric and caproic acids. PMID- 11417704 TI - Inhibition of growth of methane-producing bacteria of the ruminant forestomach by hydroxymethylglutaryl-SCoA reductase inhibitors. AB - Two hydroxymethylglutaryl-SCoA (HMG-CoA) reductase inhibitors, mevastatin and lovastatin, inhibited the in vitro growth and production of CH4 of strains of Methanobrevibacter isolated from the rumen. Mevastatin or lovastatin did not inhibit growth of species of rumen bacteria that are essential for fermenting cellulose, starch and other plant polysaccharides to acetate, propionate, and butyrate. Approximately 4 nmol of lovastatin per milliliter resulted in 50% growth inhibition of Methanobrevibacter strain ZA10 and concentrations > or =10 nmol per milliliter completely inhibited growth and CH4 formation. Results of in vitro growth studies suggest that supplementation of ruminant feeds with HMG-CoA inhibitors could decrease ruminant methane production and increase the efficiency of feed utilization by domestic ruminants. PMID- 11417705 TI - Digestion of ryegrass pasture in response to change in pH in continuous culture. AB - The ruminal pH of dairy cows fed high quality pasture is often below values recommended to optimize digestion. Four continuous culture fermenters were used to determine the pH required for the optimal digestion of pasture. High quality pasture was fermented at four controlled levels of pH (5.4, 5.8, 6.2, and 6.6) according to a 4 x 4 Latin square design. Automatic infusion of 5 M NaOH and 5 M HCl controlled pH to +/- 0.1. Digesta samples were collected during the last 3 d of each of the four 9-d experimental periods. Digestion and synthesis of microbial protein were largely insensitive to pH across a broad range of pH (5.8 to 6.6), but a large reduction in both occurred when pH was 5.4. The digestibility of pasture dry matter and synthesis of microbial protein were optimized at pH 6.35 and 6.13, respectively. The proportions of individual volatile fatty acids were not changed as pH increased. Digestion of high quality pasture in continuous culture was comparable to that predicted by the Cornell Net Carbohydrate and Protein System between pH 6.2 and 6.6. However, the model underpredicted organic matter and fiber digestibility between pH 5.4 and 5.8, compared with values obtained in continuous culture. This suggests that when ruminal pH is less than 6.2, the model may over-predict the production response to supplementation of high quality pasture with an effective fiber source. PMID- 11417706 TI - Site of nutrient digestion by dairy cows fed corn of different particle sizes or steam-rolled. AB - Five primiparous Holstein cows were cannulated in the rumen, duodenum, and ileum and were fed diets containing 50% alfalfa silage and 36.6% coarse-, medium-, or fine-ground corn (CGC, MGC, and FGC, respectively, with mean particle sizes of 4.8, 2.6, or 1.2 mm), steam-rolled corn (SRC; density of 0.53 kg/L), or a 50:50 mix of CGC and SRC (SC) to evaluate how corn processing affects site of digestibility of nonstructural carbohydrates (NSC) and neutral detergent fiber (NDF). Decreasing the particle size of corn quadratically affected true ruminal digestibility of NSC (49.8, 46.5, and 87.0%, respectively). Because of compensatory digestion postruminally, a smaller increase (from 91.3 to 98.0%) in total tract digestibility of NSC was noted as particle size decreased. A small but significant linear shift in NDF digestion from the rumen to the large intestine was detected as corn particle size decreased. The addition of SRC to CGC linearly increased true NSC digestibility in the rumen about 20 percentage units but had much smaller effects on total tract digestibility. Despite the large impact of corn processing on NSC digestibility in the rumen, flow of bacterial N to the rumen was not affected by treatment. Reducing the particle size of corn decreased the apparent escape of corn protein, but steam-rolling had no effect. Corn should be finely ground to maximize total tract OM digestibility or steam-processed to densities less than 0.53 kg/L for maximal starch digestibility. However, fine-grinding or steam-processing of corn may have only a modest impact on total tract OM digestibility. PMID- 11417707 TI - Effect of concentrate feed level in late gestation on subsequent milk yield, milk composition, and fertility of dairy cows. AB - The effects of level of concentrate feeding in late gestation on feed intake, milk yield, milk composition, and fertility in the subsequent lactation were evaluated in a randomized block design experiment involving 60 cows. Grass silage was offered ad libitum for the last 28 d of gestation either as the sole diet (OC) or supplemented with 5 kg/d of concentrates (5C). Following calving, the cows were offered the same grass silages supplemented with 7 kg/d of concentrates. For treatments OC and 5C, total dry matter intakes were 9.28 and 11.03 kg/d of dry matter, respectively, during the last 4 wk of gestation. During wk 1 to 12 of the subsequent lactation, treatment 5C increased milk fat concentration but did not alter feed intake, milk yield, or protein concentration relative to treatment OC. Treatment 5C increased the interval to first progesterone rise and the number of services per conception relative to treatment OC. Cow parity, BF depth assessed at d 28 before parturition, and treatment provided the best fit relationships for the yields of fat and fat plus protein (R2 relationships = 0.65 and 0.64, respectively) during wk 1 to 4 of lactation. It was concluded that, other than milk fat concentration, supplementation with additional concentrates in late gestation did not alter milk yield or composition and dairy cow fertility. Furthermore, despite the very large differences in cow characteristics at d 28 before parturition, there was no evidence of any interaction between treatment and specific cow characteristics on animal performance in the first 12 wk of lactation. PMID- 11417708 TI - Ruminal degradability, intestinal disappearance, and plasma methionine response of rumen-protected methionine in dairy cows. AB - Bioavailability of Met from a rumen-protected Met product was evaluated in two experiments using three ruminally and duodenally cannulated lactating (experiment 1) and nonlactating (experiment 2) dairy cows. In the first experiment, the ruminal in situ and mobile bag technique was used to assess ruminal degradability and intestinal disappearance of Met from the protected Met product. Effective ruminal degradability of Met at a ruminal outflow rate of 0.11/h was 21.7%. Combining effective ruminal degradability with intestinal digestibility yielded an estimate of Met availability of 25%. In the second experiment, designed as a 3 x 3 Latin square, Met availability was assessed by determining the response of plasma Met to supplementation of the protected Met product relative to that of duodenally administered Met. The periods were 1 wk with cows fed a meal containing 0, 20, or 63 g of protected Met on d 1 and infused intraduodenally with 10.7 g of Met on d 4. Blood was collected at various times relative to the time of oral dosing and the commencement of the duodenal infusion. Plasma Met response measured as area under the curve increased linearly with increasing protected Met. The response of plasma Met increased by 33 and 65.5% of the control values for 20 and 63 g of protected Met, respectively. Intestinal bioavailability of Met in the protected Met product ranged from 22 to 34%. PMID- 11417709 TI - Effects of abomasal infusion of long-chain fatty acids on splanchnic metabolism of pancreatic and gut hormones in lactating dairy cows. AB - Pancreatic and gut peptide hormones are potential mediators of the reduction in dry matter intake (DMI) often observed in lactating dairy cows fed supplemental fat. We investigated the effects of 7-d abomasal infusions of a rapeseed and sunflower oil mixture providing mostly unsaturated long-chain fatty acids (LCFA) on arterial concentration and splanchnic (portal-drained viscera [PDV] and liver) metabolism of insulin, pancreatic (PAN) and gut (GUT) glucagon, glucagon-like peptide-1 (7-36) amide (GLP-1), and cholecystokinin (CCK) in six cows at 55 (ELAC) and 111 (MLAC) d postpartum. Plasma flow for the PDV and liver were greater in ELAC and increased by oil infusion. Arterial concentrations of insulin and PAN were greater in MLAC, whereas arterial concentrations of GLP-1 and CCK were greater in ELAC. Abomasal oil infusion increased arterial concentration of GUT and GLP-1 but decreased arterial insulin concentration. These differences in peripheral hormone concentration were due largely to changes in their net PDV release and (or) liver removal. In addition, net liver removal of PAN was increased by oil infusion. There was no effect of oil infusion on splanchnic metabolism or arterial concentration of CCK. Lower concentrations of CCK in MLAC were attributable to net liver removal, emphasizing the importance of liver metabolism in determining peripheral concentrations of gut and pancreatic peptide hormones. Results of this study suggest a role for products of proglucagon processing (PAN, GUT, and GLP-1) as mediators of the reduction in DMI caused by postruminal supply of LCFA. PMID- 11417710 TI - Relationships of sire predicted transmitting ability for somatic cell score with measures of daughter performance. AB - The relationship between sire predicted transmitting ability (PTA) for somatic cell score (SCS) and occurrence of mastitis in daughters was characterized with 304 Holsteins first freshening since 1991 in the Virginia Tech dairy herd. No direct sire selection for PTA SCS was practiced in the herd. Linear regressions of first lactation average SCS (1.3), number of cases of mastitis (0.80), days clinical (7.0), and number of treatments for mastitis (2.0) on PTA SCS were significant. Linear regression coefficients for second and third lactations were generally positive, but like quadratic and cubic coefficients in this study, were not significant. Relationship of sire PTA SCS and measures of lifetime (84 mo opportunity) performance were determined for 2,494,195 Holsteins born between 1979 and 1987. Linear coefficients from regression on PTA SCS for number of lactations (-0.31), days of productive life (-87.0), total days in milk (-72.2), lifetime milk (-1609.1), fat (-80.7), and protein (-48.2) production, relative net income adjusted for opportunity cost for fluid markets (+13.47), first lactation average SCS (+1.00), and first lactation mature equivalent milk (+429.9) were different from zero. Partial regressions holding PTA milk or PTA protein constant were substantially larger than linear coefficients. When PTA productive life was held constant, the relationship between PTA SCS and measures of herd life and lifetime yield became positive, and relative net income adjusted for opportunity cost variables became moderately positive. One of the major expenses associated with higher susceptibility (higher PTA SCS) may be shorter herd life. PMID- 11417711 TI - The relationship between body condition score and reproductive performance. AB - The aim of this study was to investigate the relationship between measures of body condition score collected from calving until wk 26 of lactation and reproductive measures (calving interval, days to first heat, days to first service, and conception at first service). Since 1973 sires of cows at the Langhill Dairy Cattle Research Centre have been selected for either high (selection line) or average (control line) genetic merit for fat plus protein. The data included 1211 records from 534 cows calving from 1988 to 1999. At first calving, cows were randomly assigned to one of two ad libitum diets: one that was relatively high in concentrates (approximately 3000 kg/yr) and one that was relatively low in concentrates (approximately 1500 kg/yr). Selection line cows were on average thinner and lost more condition in early lactation than control line cows. Cows that lost condition, those that were thinner than average at wk 10 of lactation and those that were thinner on average over the first 10 wk, had poorer reproductive performance. This effect was greatest in the selection line. Line x diet interaction effects were not statistically significant. Genetic correlations between body condition score and reproductive measures were unfavorable and ranged from -0.04 to -0.54. The relationship between body condition score and production was strong, but, even after adjusting for yield, an unfavorable relationship still exists between body condition score and fertility. Body condition score could be used as a management and selection tool to improve reproductive performance. PMID- 11417712 TI - Detection of quantitative trait loci affecting milk production traits on 10 chromosomes in Holstein cattle. AB - Sons (n = 71 to 75) of each of six Holstein sires were genotyped at 69 microsatellite loci covering a total of 676 cM on chromosomes 3, 5, 9, 10, 13, 15, 17, 20, 23, and 26. Estimates of quantitative trait loci (QTL) effect and location were made using a least squares interval mapping approach based on daughter yield deviations of sons for 305 d milk, fat, and protein yield and fat and protein percentage. Thresholds for statistical significance of QTL effects were determined from interval mapping of 10,000 random permutations of the data across the bull sire families and within each sire family separately. Analyses combining data across sires indicated the presence of QTL affecting milk, fat, and protein yield on chromosomes 20 and 26 and a QTL affecting fat and protein percentage on chromosome 3. Analyses within each sire family separately indicated the presence of segregating QTL in at least one family on 7 of the 10 chromosomes. Statistically significant estimates of QTL effects on breeding value ranged from 438 to 658 kg of milk, from 17.4 to 24.9 kg of fat, 13.0 to 17.0 kg of protein, 0.04 to 0.17% fat, and 0.07 to 0.10% protein. PMID- 11417713 TI - Application of a structural model for genetic covariances in international dairy sire evaluations. AB - A structural model for estimation of genetic covariances in international dairy sire evaluations was compared with a standard multiple-trait mixed model using milk yield data from 13 environments (Austria, Belgium, Czech Republic, Denmark, Estonia, Finland, Israel, Switzerland and five regions of the United States). Data consisted of 2,885,117 first-lactation records from daughters of 17,867 Holstein sires. Parameters of the structural model for genetic covariances included an intercept and measures of genetic, management, and climate similarity. Genetic similarity was defined as the ratio between the number of daughters of common bulls used in a given pair of regions and the total number of daughters of all bulls. The measure of management similarity was the ratio between the absolute value of the difference in average milk yield between two regions and the sum of these averages. Climate similarity was a function of the difference in heat indices between pairs of regions. Results showed that the structural model for the genetic covariances gave more precise estimates of the latter than the standard multivariate analysis because the number of parameters to be estimated in the genetic covariance matrix was reduced from 91 to 16. A comparison of the two models using a deviance information criterion (a measure of quality of fit) showed a slight superiority for the structural model. For the latter, the posterior means of the genetic correlations between environments ranged between 0.63 and 0.98. PMID- 11417714 TI - Analysis of survival in dairy cows with supplementary data on type scores and housing systems from a region of northwest Germany. AB - In survival analysis, type traits can be included as covariates to evaluate their use as predictors for survival. One problem in such an analysis is the availability of suitable data. Whereas data on the length of productive life (LPL) of individual cows can be retrieved from milk recording data, for type traits, all cows in the population must be scored for type at least once. In the present analysis, a dataset from the Osnabruck region in northwestern Germany, which fulfilled this requirement in recent years, was used. Data consisted of 169,733 cows with information on LPL for calving years 1980 to 1996 (dataset I) and of 39,233 cows with information on LPL and type for calving years 1990 to 1996 (dataset II). A further dataset (III) contained 43,116 cows from calving years 1987 to 1996 and included information on the housing system for each herd. The basic model included stage of lactation, relative production within herd, change of herd size, and year-season as time dependent effects; age at calving as a time-independent effect; and herd-year-season and sire as random effects. Other effects (information on type, housing system) were included additionally. For data-set II, the scores for 15 linear type traits were also included as corrected phenotypic values, estimated breeding values, and residuals from a previous BLUP analysis. The package Survival Kit 3.0 was used for all analyses. The results indicate a moderate heritability of 0.17 and 0.18 for true and functional LPL (dataset I). Almost all type traits analyzed (dataset II) exceeded a 0.001 level of significance in their effect on survival. The strongest relationships between survival and type were found for udder depth, fore udder attachment, and front teat placement. The main result from the comparison of housing systems (dataset III) was that bedding has a positive effect on survival. PMID- 11417715 TI - Efficacy of a new premilking teat disinfectant containing a phenolic combination for the prevention of mastitis. AB - A teat disinfectant containing a phenolic combination was evaluated in a natural exposure study in two dairy research herds. Premilking teat disinfection was compared with a negative control using a split-udder experimental design. In both herds, premilking and postmilking teat disinfections with the phenolic combination were significantly more effective in preventing new intramammary infection (IMI) than was postmilking teat disinfection only. Clinical mastitis and new IMI by Streptococcus uberis, Streptococcus dysgalactiae, Gram-negative pathogens, and coagulase-negative Staphylococcus species were significantly lower in quarters of cows with teats predipped and postdipped than in quarters with teats postdipped only. No chapping or teat skin irritation was observed. Premilking teat disinfection with the phenolic combination in association with good udder preparation and postmilking teat disinfection can further reduce the occurrence of new IMI by numerous mastitis pathogens during lactation. PMID- 11417716 TI - Immunological parameters of Javanese carp Puntius gonionotus (Bleeker) exposed to copper and challenged with Aeromonas hydrophila. AB - This study was to determine the median lethal concentration (LC50) of copper to Javenese carp, Puntius gonionotus (Bleeker), and the immune response after the fish were exposed to sublethal levels of copper and challenged with formalin killed Aeromonas hydrophila. The LC50 of copper on P. gonionotus at 24, 48, 72, 96 and 120 h were estimated as 2.17, 0.91, 0.57, 0.53 and 0.42 mg l(-1), respectively. To determine the effect of copper on the immune system, fish were exposed for 66 days to 0.05, 0.10 and 0.15 mg Cu l(-1). After 56 days of initial exposure to copper, fish were challenged with 0.1 ml of 4.5 x 10(5) cfu ml(-1) formalin killed A. hydrophila and maintained in the same concentration of copper. After the challenge, the immune response was monitored for 2 weeks using haematological and serological assays. During the initial phase of exposure to copper, significant changes were noted in the white blood cell, lysozyme, potential killing activity, total plasma protein, total immunoglobulin and haematocrit levels between the control and treated fish. One week after challenge with A. hydrophila, there was a significant increase in the values of white blood cells, total protein and total immunoglobulin compared to the values before the challenge. However, these values were not significantly different (P>0.05) between the control and the treated fish. In contrast, NBT and lysozyme assays exhibited a significant difference (P<0.05) in fish exposed to 0.10 mg Cu l(-1) (0.525 +/- 0.17; 24.42 +/- 3.35 x 10(2) micromg ml(-1)) and 0.15 mg Cu 1(-1) (0.536 +/- 0.19; 21.78 +/- 1.29 x 10(2) micromg ml(-1)) compared to the control (0.746 +/- 0.31; 30.73 +/- 5.42 x 10(2) micromg ml(-1)) after the bacterial challenge (day 61). There was however no significant difference (P>0.05) in NBT and lysozyme levels in fish exposed to lower level of copper (0.05 mg Cu l(-1)), suggesting the absence of immunosuppressive effects at lower level of exposure. PMID- 11417717 TI - Vitamin E increases natural cytotoxic activity in seabream (Sparus aurata L.). AB - The natural cytotoxic activity of head-kidney leucocytes from gilthead seabream (Sparus aurata L.), after in vitro and in vivo vitamin E treatment, against tumor cells was studied by flow cytometry. Leucocytes were incubated in culture medium with different vitamin E supplementations (0.01-10 microg ml(-1)) for 6, 24 or 48 h and the results demonstrate that all the assayed vitamin E supplementations significantly enhanced the natural cytotoxic activity of leucocytes. To determine the effect of a high dietary level of vitamin E on this activity, fish were fed with 0 (control), 600, 1200 or 1800 mg of vitamin E supplementation kg(-1) diet for 2, 4 or 6 weeks. After 2 and 4 weeks of treatment, the natural cytotoxic activity was significantly enhanced at the highest (1.8 g kg(-1) diet) and lowest (600 mg kg(-1) diet) vitamin E supplement dosage, respectively. No effect of the vitamin E supplemented diet on seabream leucocyte natural cytotoxic cell activity was observed after 6 weeks of treatment. PMID- 11417718 TI - Immunomodulatory effects of dietary intake of chitin on gilthead seabream (Sparus aurata L.) innate immune system. AB - To determine the effects of chitin (poly [1-->4]-beta-N-acetyl-D-glucosamine) on the innate immune response of gilthead seabream (Sparus aurata L.), fish were fed diets containing 0 mg (control), 25, 50 or 100 mg kg(-1) chitin for 2, 4 and 6 weeks. Lysozyme and natural haemolytic complement activities, together with head kidney leucocyte respiratory burst, phagocytic and cytotoxic activities, were studied at each of the assayed times. Lysozyme activity was unaffected by the administration of chitin. The innate humoral and cellular immune response activities assayed were enhanced by the dietary intake of chitin, each increasing at a different time: natural haemolytic complement activity and cytotoxic activity after 2 weeks of treatment, respiratory burst activity from 4 weeks and phagocytic activity after 6 weeks, although, unlike the other activities, no statistically significant differences were observed in the first. The results indicate that chitin increases the activity of the seabream innate immune system, and its use as an immunostimulant is discussed, especially with regards to the protective role. PMID- 11417719 TI - Kinetics of juvenile sea bass (Dicentrarchus labrax, L.) systemic and mucosal antibody secreting cell response to different antigens (Photobacterium damselae spp. piscicida, Vibrio anguillarum and DNP). AB - The ELISPOT assay was used to measure the number of specific antibody secreting cells (ASC) induced during the primary and secondary immune responses in the spleen, head kidney and gut of juvenile (5 g) sea bass (Dicentrarchus labrax) to bacterial (Vibrio anguillarum and Photobacterium damselae ssp. piscicida) and hapten dinitrophenyl-conjugated to keyhole limpet haemocyanin (DNP-KLH) antigens administered intraperitoneally. High variability among individuals was observed at each sampling day. All fish were bath vaccinated to V. anguillarum at an earlier stage (2 g) in the farm of origin prior to the development of the experiments, and therefore only secondary and tertiary responses were measured in the group immunised with this bacterium. Significant differences to the controls were observed in the primary responses of the head kidney and the spleen to P. damselae ssp. piscicida and DNP, respectively. Frequency analysis of the production of ASC suggests that significant responses in the gut might be masked by the high error variance. The peak of the primary response was observed 4 days earlier to DNP (18-20 days post-immunisation) and it was significantly higher than the response to P. damselae ssp. piscicida. Higher numbers of ASC were observed in the secondary responses of the head kidney and spleen, although they were not statistically significantly different from the primary levels, probably due to the high error variance as supported by the frequency analysis. Nevertheless, together with a faster response (peak at 7 days post-immunisation), the data suggest that memory formation had occurred. Additionally, the data suggest that some suppression of the secondary immune response in the gut might have occurred. The head kidney appears to produce the highest number of specific ASC of the organs tested. It appears that sea bass show a relatively fast but short duration antibody response. PMID- 11417720 TI - The kinetics of the hypoferraemic response and changes in levels of alternative complement activity in diploid and triploid Atlantic salmon, following injection of lipopolysaccharide. AB - To study any possible effects of triploidy on the kinetics of the response of two non-specific disease factors, full sibling diploid and triploid Atlantic salmon were injected intraperitoneally with either lipopolysaccharide (1 mg kg(-1) body weight) or saline. Individually marked fish were repetitively blood sampled for up to 19 days. Total serum protein concentrations remained constant throughout the experiment indicating that the sampling regime did not cause haemodilution. The alternative complement pathway activity (measured by the titre of haemolytic activity against rabbit erythrocytes) in the serum of saline injected fish remained constant but in LPS-injected fish it fell to barely detectable levels 2 days after injection, but recovered to pre-treatment levels by about day 5. Triploid fish took slightly longer to reach full recovery levels than diploids. All groups of fish showed a hypoferraemic response, suggesting that the sampling regime was at least partially responsible. However, the response was more rapid and pronounced in the LPS-injected fish. In the latter, serum iron concentrations decreased to very low levels by day 2 post-injection in the diploid fish and by day 3 in the triploid fish. Pre-treatment iron levels were re-established by about 15 days post-injection in all groups. The data show only slight differences between the diploid and triploid fish, but the longer time taken for the triploids to recover complement activity and the slower onset of the hypoferraemic response following injection of LPS, suggest that they may be at a disadvantage compared with their diploid siblings in their defence against bacterial infections. PMID- 11417721 TI - Effect of ascorbic acid on the immune response of the catfish, Mystus gulio (Hamilton), to different bacterins of Aeromonas hydrophila. AB - Ascorbic acid is one of the essential vitamins for normal physiological activities of any organism. The present study demonstrates an immunostimulatory effect of vitamin C on the humoral and cell mediated immunity of the bagrid catfish, Mystus gulio, determined using different bacterins of Aeromonas hydrophila. Humoral as well as cell mediated immune responses were elucidated in the vitamin supplemented, vaccinated and unvaccinated groups. The vitamin supplemented lipopolysaccharide (LPS) vaccinated group exhibited greater immune (both humoral and cell mediated) responses than its formalin killed (FK) and heat killed (HK) bacterin vaccinated counterparts. Nevertheless, in the challenge study, the relative percent survival (RPS) was found to be the same for both FK and LPS immunised vitamin treated groups while lower for the HK immunised vitamin treated group. PMID- 11417722 TI - The analysis of expressed genes in the kidney of Japanese flounder, Paralichthys olivaceus, injected with the immunostimulant peptidoglycan. AB - Immunostimulants are widely used in aquaculture, but there are few reports on the genes that are expressed by their stimulation. Therefore, in this study, expressed genes in the kidney of Japanese flounder Paralichthys olivaceus injected with the immunostimulant peptidoglycan were analysed. The results of single-pass sequencing of ESTs from 198 clones (AU090255-AU090451, AU090935) from kidney cDNA are presented. Sequences of the cDNA clones were compared with sequences in the GenBank database. One hundred and six clones (53.5%) appeared to be completely unknown and are likely to represent newly described genes, whereas 92 clones (46.5%) were identified based on matches to sequences in the database. The results contain the genes such as alpha globin (AU090287), several ribosomal proteins (AU090-263, 274, 299, 351, 365, 375, 377, 382, 434, 445), heat shock protein 90 (AU090374) and cytochrome oxidase subunit (AU090385). Immune related cDNAs identified from the kidney were immunoglobulin heavy (AU090291) and light chain (AU090352), beta2-microglobulin (AU090280), macrophage inflammatory protein 1-alpha precursor (AU090535), thymosin beta-10 (AU090391), lysozyme (AU090322) and MHC class IIalpha (AU090435). It is possible that expression of macrophage inflammatory protein 1-alpha results in macrophage activation as a consequence of peptidoglycan treatment. PMID- 11417723 TI - Evidence for the immune recognition of two haemolysins of Renibacterium salmoninarum by fish displaying clinical symptoms of bacterial kidney disease (BKD). PMID- 11417724 TI - Kentucky veterinarians investigate mysterious foal death syndrome. PMID- 11417725 TI - PAVE program targeted in one of six resolutions. PMID- 11417726 TI - A close call for the National Zoo's Conservation and Research Center. PMID- 11417727 TI - No disappointments [interview by R. Scott Nolen]. PMID- 11417728 TI - Computers--learning from loss. PMID- 11417729 TI - Disagree with use of muscle relaxant before euthanasia. PMID- 11417730 TI - Food animal welfare and the use of bovine somatotropin. PMID- 11417731 TI - ECG of the month. Cardiac tamponade secondary to A. niger-induced constrictive pericarditis. PMID- 11417732 TI - Penicillins and beta-lactamase inhibitor combinations. AB - beta-Lactamase production by bacteria continues to be one of the main mechanisms of bacterial resistance to beta-lactam antibiotics, and it seems likely to remain so. beta-Lactamase inhibitors provide 1 strategy to overcome this mechanism of bacterial resistance. Although 3 beta-lactamase inhibitor/antibiotic combinations are currently available, only 1 is approved for veterinary use. Because the beta lactamase inhibitor must be present concurrently with the antibiotic for synergistic activity, it is important to consider the pharmacokinetic profile of these drugs in combination. These combinations were developed and optimized for human patients, so it is unlikely that they would achieve the ideal plasma and tissue concentrations and ratios in veterinary patients. Indeed, several differences in pharmacokinetic variables of beta-lactam antibiotic/beta-lactamase inhibitor agents have been described in dogs, compared with people. Such pharmacokinetic differences should be considered when interpreting in vitro susceptibility results in veterinary species, because these tests use ratios of drug that were established for humans. The beta-lactamase inhibitors represent a successful example of targeted drug development. However, the currently available inhibitors are active primarily against class-A beta-lactamases. Because the frequency with which class-C beta-lactamases are recognized is rapidly increasing in human isolates, and because beta-lactamase enzymes continue to evolve, new beta-lactamase inhibitors will need to be developed to target these enzymes. PMID- 11417733 TI - Epidemiologic features of pet evacuation failure in a rapid-onset disaster. AB - OBJECTIVE: To describe epidemiologic features of pet evacuation failure after a hazardous chemical spill in which residents had no warning and only a few hours notice to evacuate. DESIGN: Cross-sectional study. SAMPLE POPULATION: Pet-owning households that evacuated from a hazardous chemical spill with (n = 119) or without (122) their pets. Procedures-Evacuees were surveyed by mail. RESULTS: 261 of 433 (60.3%) dogs and cats in 241 households were not evacuated. Of the 241 households, 119 (49.4%) evacuated with their pets, 98 (40.7%) evacuated without them but later attempted to rescue them, and 24 (10.0%) neither evacuated their pets nor attempted to rescue them. Pet evacuation failure was most common in households that thought the evacuated area was safe for pets. Risk of pet evacuation failure increased in households with many animals, low pet attachment and commitment scores, and low levels of preparedness. Cat evacuation failure was associated with not having cat carriers. Nearly 80% of households that evacuated with their pets found accommodation with friends and family. CONCLUSIONS AND CLINICAL RELEVANCE: Pet evacuation failure was common and jeopardized pets' health and well-being. Logistical challenges to transporting pets were substantial contributors to pet evacuation failure, whereas not knowing where to house a pet was only a minor concern. Most pet owners seemed self-reliant and acted appropriately towards their pets. Such self-reliant behavior by pet owners should be encouraged prior to disasters as part of an evacuation plan for households. PMID- 11417734 TI - Risk factors for pet evacuation failure after a slow-onset disaster. AB - OBJECTIVE: To determine risk factors for pet evacuation failure during a flood. DESIGN: Cross-sectional survey. SAMPLE POPULATION: 203 pet-owning households in a flooded region. PROCEDURES: Persons under evacuation notice because of a flood were interviewed by use of a random telephone survey. RESULTS: 102 households evacuated with their pets, whereas 101 households evacuated without their pets. Low pet attachment and commitment scores were significantly associated with a greater chance of pet evacuation failure. Risk of pet evacuation failure and lower attachment and commitment scores were also associated with pet management practices prior to the disaster, such as dogs being kept outdoors most of the time or owners not having carriers for their cats. More than 90% of owners made housing arrangements for their pets without assistance. CONCLUSIONS AND CLINICAL RELEVANCE: Predictors of pet evacuation failure are usually present before a disaster strikes and are potentially modifiable. Mitigation of pet evacuation failure should focus on activities that reinforce responsible pet ownership and strengthen the human-animal bond, including socializing dogs, attending dog behavior training classes, transporting cats in nondisaster times, and seeking regular preventive veterinary care. Most pet owners are self-reliant in disasters, and this behavior should be encouraged. PMID- 11417735 TI - Salmonella control and quality assurance at the farm end of the food safety continuum. PMID- 11417736 TI - Dog bites to humans--demography, epidemiology, injury, and risk. PMID- 11417737 TI - Epidemiologic characteristics of rabies virus variants in dogs and cats in the United States, 1999. AB - OBJECTIVE: To evaluate epidemiologic features of rabies virus variants in dogs and cats in the United States during 1999 and assess the role of bat-associated variants. DESIGN: Epidemiologic survey. SAMPLE POPULATION: Rabies viruses from 78 dogs and 230 cats. PROCEDURE: Brain specimens from rabid dogs and cats were submitted for typing of rabies virus. Historical information, including ownership and vaccination status, was obtained for each animal. Specimens were typed by use of indirect fluorescent antibody assay or reverse transcriptase polymerase chain reaction assay and nucleotide sequence analysis. RESULTS: Nearly all animals were infected with the predicted terrestrial rabies virus variant associated with the geographic location of the submission. A bat-associated variant of rabies virus was found in a single cat from Maryland. More than half (53%) of submitted animals were classified as owned animals, and most had no known history of vaccination. One vaccination failure was reported in a dog that did not receive a booster dose of rabies vaccine after exposure to a possibly rabid animal. CONCLUSIONS AND CLINICAL RELEVANCE: Bat-associated rabies virus variants were not a common cause of rabies in dogs and cats during 1999. Vaccine failures were uncommon during the study period. Because most rabid dogs and cats were unvaccinated and were owned animals rather than strays, educational campaigns targeting owners may be useful. PMID- 11417738 TI - Use of a polymerase chain reaction assay for detection of Haemobartonella canis in a dog. AB - A polymerase chain reaction (PCR) originally developed for detection of Haemobartonella felis in cats was successfully used for detection of H canis in an 8-year-old spayed Great Dane. The dog had been splenectomized and was undergoing immunosuppressive chemotherapy at the time of diagnosis. Sequence analysis of the 16S ribosomal RNA gene revealed that the Haemobartonella spp infecting this dog was 97% homologous to the sequence previously reported for the Ohio strain of H felis. Clinical and hematologic abnormalities as well as identification of the organisms by use of light and electron microscopy supported the diagnosis of H canis. The PCR assay used for detection of H felis may be useful for the detection of H canis in dogs prior to blood donation, splenectomy, or treatment with immunosuppressive drugs. PMID- 11417739 TI - Succimer for treatment of lead toxicosis in two cats. AB - Two cats from a single household were examined because of neurologic abnormalities suggestive of central vestibular disease. The owner had been renovating her 150-year-old house for the preceding 3 months, and renovations included chipping and sanding of old paint from windowsills and stair railings. Lead toxicosis was diagnosed on the basis of history and concentrations of lead in blood and urine. Both cats were treated with succimer. Treatment was not associated with any adverse effects, and both cats recovered completely. Ingestion of old paint from house renovations is the most common source of lead exposure in cats. Owners of cats with gastrointestinal tract or neurologic abnormalities should be questioned specifically about house renovations. PMID- 11417740 TI - Outcome of and postoperative complications in dogs undergoing surgical treatment of laryngeal paralysis: 140 cases (1985-1998). AB - OBJECTIVE: To compare outcomes of various surgical techniques for treatment of laryngeal paralysis in dogs and determine incidence and risk factors for development of postoperative complications. DESIGN: Retrospective study. ANIMALS: 140 dogs undergoing surgical treatment for laryngeal paralysis at a veterinary teaching hospital between 1985 and 1998. PROCEDURE: Data were analyzed to determine outcome and factors influencing outcome and development of complications. Kaplan-Meier curves were constructed for survival analysis. RESULTS: Postoperative complications were documented in 48 (34.3%) dogs; 20 (14.3%) dogs died of related causes. Aspiration pneumonia was the most common complication (33; 23.6%). Seven dogs died of aspiration pneumonia > 1 year after surgery. Dogs that underwent bilateral arytenoid lateralization were significantly more likely to develop complications and significantly less likely to survive than were dogs that underwent unilateral arytenoid lateralization or partial laryngectomy. Factors that were significantly associated with a higher risk of dying or of developing complications included age, temporary tracheostomy placement, concurrent respiratory tract abnormalities, concurrent esophageal disease, postoperative megaesophagus, concurrent neoplastic disease, and concurrent neurologic disease. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that surgical repair of laryngeal paralysis may be associated with high postoperative complication and mortality rates. Surgical technique and concurrent problems or diseases increased the risk of complications. Dogs appeared to have a life-long risk of developing respiratory tract complications following surgical correction. PMID- 11417741 TI - Prevalence and clinical implications of anthelmintic resistance in cyathostomes of horses. AB - OBJECTIVE: To determine the prevalence and clinical implications of anthelmintic resistance in cyathostomes of horses. DESIGN: Prospective study. ANIMALS: 80 horses on 10 farms in a 5-county region of northeast Georgia. PROCEDURE: On each farm, horses were stratified in descending order according to pretreatment fecal egg count (FEC), blocked into groups of 4, and then randomly assigned to 1 of 4 treatment groups: no treatment (controls), and treatment with pyrantel pamoate, fenbendazole, or ivermectin. Fecal samples were collected 24 hours prior to treatment and 2, 4, and 6 weeks after treatment for determination of FEC. Mean percentage of reduction in FEC was then calculated for each treatment group. For horses from each farm, the efficacy of each anthelmintic was categorized on the basis of mean percentage of reduction in FEC at 2 weeks after treatment (< 80% reduction = ineffective; 80 to 90% reduction = equivocal; and > 90% reduction = effective). RESULTS: Pyrantel pamoate was effective at reducing FEC in horses from 7 farms, ineffective in horses from 2 farms, and equivocal in horses from 1 farm. Fenbendazole was ineffective at reducing FEC in horses from 9 farms and equivocal in horses from 1 farm. Ivermectin was effective at reducing FEC in horses from all 10 farms. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that cyathostome resistance to fenbendazole is highly prevalent, and resistance to pyrantel pamoate is high enough to warrant concern. Resistance to ivermectin was not detected. On the basis of these data, it appears that ivermectin continues to be fully effective in horses. However, too few farms were used in this study to determine the prevalence of cyathostome resistance to ivermectin. Therefore, the efficacy of ivermectin should continue to be monitored closely. PMID- 11417742 TI - Efficacy of salmeterol xinafoate in horses with recurrent airway obstruction. AB - OBJECTIVE: To determine the onset, magnitude, and duration of bronchodilation after administration of aerosolized salmeterol xinafoate in horses with recurrent airway obstruction. DESIGN: Randomized controlled study ANIMALS: 6 horses with recurrent airway obstruction. Procedure Horses received aerosolized salmeterol (210 microg) or no treatment, using a crossover design. Salmeterol was administered, using a mask designed for aerosol delivery in horses. Subjective rating of airway obstruction (RAO), maximal change in pleural pressure (deltaPplmax), and pulmonary resistance (RL) were determined at baseline; 5, 15, and 30 minutes; and 1, 2, 4, 6, 8, 10, and 12 hours after administration of salmeterol and in horses that did not receive treatment. RESULTS: The deltaPpl and RL were improved 15 minutes through 6 hours after administration of salmeterol, compared with values obtained from horses receiving no treatment. The RAO was improved 15 minutes through 2 hours after administration of salmeterol. The maximal response to salmeterol was evident 30 to 60 minutes after administration and was characterized by a 59 + 19% decrease in deltaPpl and a 56 +/- 13% decrease in RL. The deltaPpl and RL were not different from baseline values 8 hours after salmeterol administration. CONCLUSIONS AND CLINICAL RELEVANCE: Duration of action of salmeterol in these horses was approximately 6 hours. Maximal bronchodilation was somewhat delayed (30 to 60 minutes), and the magnitude of response was similar to that of short-acting beta2-adrenergic agonists. Salmeterol provides moderately sustained bronchodilation in horses with recurrent airway obstruction and may be an effective drug for long-term control of this condition. PMID- 11417743 TI - Fracture repair of the distal portion of the radius by use of a condylar screw implant in an adult horse. AB - An 8-year-old American Quarter Horse gelding was evaluated because of an open fracture involving the left radius. The horse had fallen during training and became immediately non-weight-bearing in the left forelimb. On initial evaluation, the horse was unable to bear weight on that limb; radiography revealed a long oblique fracture of the distal metaphysis of the radius with minimal displacement of the fracture fragments. Because of the configuration of the fracture, we recommended surgical intervention with internal fixation. A condylar screw implant and 4.5-mm broad dynamic compression plate were applied to the medial and dorsolateral aspects of the radius, respectively. The horse recovered in a sling and full-limb bandage. Six months after discharge, the horse was reevaluated because of a grade 4/5 lameness in the same limb. Palpation revealed signs of severe pain over the distomedial aspect of the radius. Radiography of the left radius revealed severe osteolysis beneath the distal aspect of the condylar screw implant. Surgical removal of the medial plate was performed. Sixteen months after the initial fracture repair, the horse had returned to light training without signs of lameness. Removal of the dorsal plate may be indicated if this horse is to return to aggressive training or becomes lame in the left forelimb. PMID- 11417744 TI - Neodymium:yttrium-aluminum-garnet laser ablation of a urethral web to relieve urinary outflow obstruction in a horse. AB - An 8-year-old Hanoverian gelding was examined because of urine dribbling and urethral obstruction. Mild proprioceptive deficits of the left hind limb were evident during neurologic examination. Ultrasonography per rectum revealed dilatation of the pelvic portion of the urethra. Endoscopy of the urethra revealed 2 webs of tissue: 1 was located 10 cm proximal to the external urethral opening; the other was located 65 cm proximal to the external urethral opening and prevented passage of the endoscope into the urinary bladder. The mass was ablated with a neodymium:yttrium-aluminum-garnet laser, using a transendoscopic noncontact technique. On follow-up examination 6 months after laser surgery, an endoscope could easily be passed into the bladder, and no urethral web was seen. The horse was able to void a stream of urine but continued to dribble urine intermittently. The proximal location of the urethral lesion in this horse would have made use of traditional surgical methods problematic, whereas transendoscopic laser photoablation was easy and effective. PMID- 11417745 TI - Efficacy of an inactivated respiratory syncytial virus vaccine in calves. AB - OBJECTIVE: To determine whether an inactivated bovine respiratory syncytial virus (BRSV) vaccine would protect calves from infection with virulent BRSV. DESIGN: Randomized controlled trial. ANIMALS: 27 nine-week-old calves seronegative for BRSV exposure. PROCEDURE: Group-1 calves (n = 9) were not vaccinated. Group-2 calves (n = 9) were vaccinated on days 0 and 21 with an inactivated BRSV vaccine containing a minimum immunizing dose of antigen. Group-3 calves (n = 9) were vaccinated on days 0 and 21 with an inactivated BRSV vaccine containing an amount of antigen similar to that in a commercial vaccine. All calves were challenged with virulent BRSV on day 42. Clinical signs and immune responses were monitored for 8 days after challenge. Calves were euthanatized on day 50, and lungs were examined for lesions. RESULTS: Vaccination elicited increases in BRSV-specific IgG and virus neutralizing antibody titers and in production of interferon-gamma. Virus neutralizing antibody titers were consistently less than IgG titers. Challenge with BRSV resulted in severe respiratory tract disease and extensive pulmonary lesions in control calves, whereas vaccinated calves had less severe signs of clinical disease and less extensive pulmonary lesions. The percentage of vaccinated calves that shed virus in nasal secretions was significantly lower than the percentage of control calves that did, and peak viral titer was lower for vaccinated than for control calves. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the inactivated BRSV vaccine provided clinical protection from experimental infection with virulent virus and decreased the severity of pulmonary lesions. Efficacy was similar to that reported for modified-live BRSV vaccines. PMID- 11417746 TI - Bone metastasis model with multiorgan dissemination of human small-cell lung cancer (SBC-5) cells in natural killer cell-depleted SCID mice. AB - Lung cancer is commonly associated with multiorgan metastasis, and bone is a frequent metastatic site for lung cancer. Nevertheless, no bone metastasis model of lung cancer with multiorgan dissemination is available, which could provide opportunity to study the molecular pathogenesis. We examined the abilities of eight human lung cancer cell lines injected intravenously into natural killer (NK) cell-depleted SCID mice to generate metastatic nodules in bone and multiple organs, and explored the correlation of the parathyroid hormone-related protein (PTHrP) with the bone metastasis. Although all the small-cell carcinoma cell lines (SBC-5, SBC-3, SBC-3/ADM, H69, H69/VP) formed metastatic nodules in multiple organs (liver, kidney, and lymph nodes), only SBC-5 cells reproducibly developed bone metastases. Squamous cell carcinoma (RERF-LC-AI) cells metastasized mainly into the liver and kidneys, whereas adenocarcinoma (PC-14, A549) mainly produced colonies in the lungs. As assessed by X-ray photography, the osteolytic bone metastases produced by SBC-5 cells were detected as early as on day 28, and all recipient mice developed bone metastasis by day 35. The expression of PTHrP in eight cell lines was directly correlated with the formation of bone metastasis. No correlation was observed between the formation of bone metastasis and the expression of other metastasis-related cytokines (IL 1, IL-6, IL-8, IL-10, IL-11, TNF-alpha, VEGF, M-CSF). Consistent with the formation of bone metastasis by SBC-5 cells, the levels of PTHrP and calcium in the mouse serum were increased in a time-dependent manner, suggesting that PTHrP produced by human lung cancer may play a crucial role in the formation of bone metastasis and hypercalcemia. These findings indicate that a bone metastasis model of SBC-5 cells may be useful for clarifying the molecular aspects of the metastatic processes in different organ microenvironments and the development of therapeutic modalities for lung cancer patients with bone metastases. PMID- 11417747 TI - Indoloquinoxaline compounds that selectively antagonize P-glycoprotein. AB - Tumor cells often develop drug resistance through overexpression of membrane transport proteins that effectively efflux anticancer agents. The pharmacologies of the two best-studied transporters, P-glycoprotein (Pgp) and MRP1, are partially overlapping but distinct. To improve the therapeutic potential of drug resistance reversing agents, we have developed a program to identify compounds with selectivity for Pgp or MRP1. Screening of a commercial library of compounds identified indoloquinoxaline compounds with transporter selectivity, and certain examples were synthesized and further evaluated. 1,4-Dibutoxy-6H-indolo[2,3 b]quinoxaline and 4,7-dibutoxy-2,3-dihydrobenzimidazole-2-spiro-3-indolin-2-one were synthesized by condensation of 3,6-dibutoxy-1,4-diaminobenzene and isatin. Neither compound was cytotoxic to MCF-7 cells, nor did either one affect the sensitivity of MCF-7/VP or HL-60/ADR cells at doses up to at least 20 microM, indicating that they do not antagonize MRP1. In contrast, each compound, at doses as low as 0.25 microM, sensitized NCI/ADR cells to vinblastine, actinomycin D, Taxol, and doxorubicin, indicating that they effectively reverse Pgp-mediated multidrug resistance (MDR). Furthermore, the compounds sensitized two additional cell lines that overexpress Pgp to this panel of anticancer drugs. However, these compounds did not affect the sensitivities of MCF-7 or T24 cells to these cytotoxic drugs, and did not alter the sensitivities of any of the tested cell lines to cisplatin or 5-fluorouracil. Both compounds enhanced the intracellular accumulation of [3H]vinblastine by NCI/ADR cells, but did not inhibit photoaffinity labeling of Pgp by [3H]azidopine at concentrations up to at least 100 microM. Therefore, these novel nontoxic indoloquinoxalines selectively sensitize Pgp-overexpressing cells to drugs that are subject to transport by this protein, without modulating the sensitivities of MRP1-overexpressing or non-Pgp cells to cytotoxic drugs. Because of this transporter selectivity, we predict that these compounds will be effective MDR modulators in vivo. PMID- 11417748 TI - Differences in the induction of DNA damage, cell cycle arrest, and cell death by 5-fluorouracil and antifolates. AB - Thymidylate synthase (TS) is an important target for chemotherapy and can be inhibited by 5-fluorouracil (5-FU) and the antifolates, AG337 (Nolatrexed) and multitargeted antifolate (MTA or Pemetrexed). In addition, 5-FU can be incorporated into RNA and DNA, and MTA can inhibit two other enzymes. It is, however, unclear to what extent these differences in drug action will influence activation of downstream mechanisms mediated via TS inhibition. Therefore, two human colon cancer cell lines, WiDr and Lovo, with a different clonogenic origin, were treated with equitoxic concentrations of 5-FU, AG337, and MTA to determine the induction of DNA damage, cell cycle arrest, downstream protein expression, and cell death. At these concentrations, the specific TS inhibitor AG337 induced more DNA damage (up to 20%) than MTA and 5-FU. FACS analysis showed that all drugs induced S phase arrest in Lovo and WiDr that was most pronounced after 5-FU and AG337 exposure (50-70%). Western blotting showed that p53 induction was not detectable in mutant (mt) p53 WiDr and increased much earlier in wild-type (wt) Lovo cells after 5-FU and MTA (24 h) than after AG337 exposure (72 h). In contrast to 5-FU-treated Lovo cells, the bcl-2/bax ratio decreased after antifolate exposure. Nevertheless, both 5-FU and antifolates induced similar amounts of cell death (up to 60%). These results demonstrate that in human colon cancer cells differences in downstream events between AG337 and 5-FU or MTA are related to the additional effects of 5-FU and MTA, which are not associated with TS inhibition. PMID- 11417749 TI - Angiogenic and immune parameters during recombinant interferon-alpha2b adjuvant treatment in patients with melanoma. AB - As an adjuvant therapy for patients with high risk of recurrent melanoma, high dose interferon (IFN)-alpha2b therapy has been shown to have some efficacy. We examined 22 patients with resected melanoma who were treated with repeated injections of recombinant IFN-alpha2b during the treatment. Both angiogenic and immune parameters were measured. White blood cells (WBCs) and lymphocyte numbers, lymphocyte subpopulations, serum concentrations of IFN-alpha and anti-IFN-alpha antibodies, and the serum vascular endothelial growth factor (VEGF), interleukin (IL)-8, and basis fibroblast growth factor (bFGF) concentrations were determined over time in resected, recurrence-free patients with American Joint Committee on Cancer (AJCC) stage III melanoma with one or less (LN+ < or = 1, n = 7) or more than one (LN+ > 1, n = 8) lymph nodes involved, and AJCC stage IV resected disease (n = 7). Follow-up and recurrence-free intervals were longer in stage III (LN+ < or = 1) patients compared with stage IV patients (P < 0.05). The number of WBCs and lymphocytes decreased during the treatment for all patient groups (P < 0.001). In addition, percentages of CD8 and CD20 were higher in stage IV patients than in stage III (LN+ > 1) and stage III (LN+ < or = 1) patients at the beginning of therapy (P < 0.05). A significant increase in the percentage of CD20+ cells, mostly B lymphocytes, was observed in the stage III (LN+ > 1) and stage III (LN+ < or = 1) patients over time but not in stage IV patients (P < 0.001). Low IL-8 and bFGF concentrations at the beginning of therapy were associated with significantly longer recurrence-free survival (P < 0.05). These results warrant a larger trial to determine if the differences observed in patients before treatment can provide prognostic markers in patients receiving IFN-alpha2b therapy. PMID- 11417750 TI - Cardiac and renal dysfunction in chronic heart failure: relation to neurohumoral activation and prognosis. AB - BACKGROUND: In chronic heart failure (CHF), cardiac dysfunction is considered the major determinant of neurohumoral activation but the role of renal impairment has not been defined. We investigated the relationship between both cardiac and renal dysfunction and neurohumoral activation, and their possible influence on prognosis. METHODS: Hemodynamics, renal function, plasma neurohormones, and long term follow-up were evaluated in 148 CHF patients, grouped according to systolic volume index (SVI) and serum creatinine (CRE) values: SVI > 28 mL/m2 and CRE < 1.5 mg/dL (group I, n = 55), SVI < 28 mL/m2 and CRE < 1.5 mg/dL (group II, n = 37), SVI > 28 mL/m2 and CRE > 1.5 mg/dL (group III, n = 25), SVI < 28 mL/m2 and CRE > 1.5 mg/dL (group IV, n = 31). RESULTS: Neurohormones progressively increased from Group I through IV and correlated with both cardiac and renal function. The hemodynamic pattern was similar in patients with normal or abnormal renal function, whereas neurohormones were only moderately increased in the former group and markedly increased in the latter group. Long-term survival progressively decreased from Group I through IV and was significantly poorer in patients with renal dysfunction. CONCLUSIONS: Our study confirms that, in CHF, neurohumoral activation is strictly related to long-term survival and that many factors contribute to its development and progression; among these, cardiac and renal dysfunction seem to play a major role. PMID- 11417751 TI - Effect of bone biopsy in guiding antimicrobial therapy for osteomyelitis complicating open wounds. AB - BACKGROUND: Osteomyelitis associated with infected overlying wounds represents a difficult diagnostic and therapeutic problem; bone biopsies can be done during debridement of the overlying wounds, but it is unclear how often the results of these bone cultures actually affect subsequent antibiotic decisions. The present study was undertaken to evaluate the usefulness of bone biopsies in guiding antibiotic therapy for this type of osteomyelitis. METHODS: Culture results of 44 bone biopsies taken during surgical debridement in 41 patients over the period from June 1994 to August 1998 were compared with those from the overlying wounds to determine whether the data affected the subsequent choice of antibiotics. The study design was that of a retrospective chart review in which the standard operative and microbiological procedures in place at the Milwaukee Veterans Affairs Medical Center were used. RESULTS: Sixty-one wound and 55 bone isolates were obtained during this study. Thirty-one isolates were found in bone, but not the overlying wound; diphtheroids were the most common organism obtained in this fashion. Correlation between wound and bone isolates was generally poor. Antibiotics were subsequently changed in 20 of the 44 cases after results of the bone biopsy became known, with the bone isolates already being covered in 10 cases and the bone biopsy results ignored in 14 cases. CONCLUSION: Because bone biopsy results seem to aid in tailoring antibiotic therapy in almost half the cases when bone is sampled during wound debridement surgery, this technique may be very helpful in certain cases and should be regularly undertaken when these procedures are carried out. PMID- 11417752 TI - Zoster in patients infected with HIV: a review. AB - Varicella-zoster virus (VZV), a member of the human herpesvirus family, causes childhood chickenpox (varicella), becomes latent in sensory ganglia, and reactivates years later in immunocompromised and elderly persons to produce shingles (herpes zoster). Early in the AIDS epidemic, zoster was noted in adults and children infected with HIV. Severe and debilitating zoster-associated dermatological, ophthalmic, and neurological complications may occur in patients infected with HIV. Antiviral therapy can modify the duration of zoster and alleviate its attendant complications. Varicella vaccine may boost the immunity and prevent virus reactivation. VZV immune globulin (VZIG) prevents or modifies clinical illness in persons who have been exposed to varicella or zoster. PMID- 11417753 TI - Impaired renal autoregulation: implications for the genesis of hypertension and hypertension-induced renal injury. AB - In summary, autoregulation of the renal vasculature provides a mechanism by which renal function is maintained relatively constant despite variations in systemic blood pressure. This system also provides a means for changes in blood pressure to occur without causing inappropriate alterations in urinary NaCl excretion. Alterations in the autoregulatory response can have clinical consequences. Increased activity of the TGF mechanism may be causally related to the development of some forms of hypertension. Decreased activity of TGF or an impaired myogenic response may help explain the increased susceptibility that certain patient groups exhibit toward hypertension-induced renal injury. The aggressive treatment of hypertension in patients with impaired renal autoregulation may be associated with an increase in the serum creatinine concentration. As long as this increase is neither excessive nor progressive, physicians should not be dissuaded from trying to achieve newly established blood pressure goals. PMID- 11417754 TI - Renal scars masquerading as complex masses in a patient with vesicoureteral reflux nephropathy. AB - Vesicoureteral reflux can lead to chronic pyelonephritis, renal scarring, and renal failure. We present a case of renal scarring masquerading as bilateral, complex renal masses. A 35-year old woman who was diagnosed with vesicoureteral reflux as a child presented for evaluation of recently developed hypertension and an abnormal renal ultrasound. Her serum creatinine level was 2.5 mg/dL and she had subnephrotic-range proteinuria. A renal sonogram showed small, echogenic kidneys and bilateral complex renal masses of 3.8 (right) and 4.4 (left) cm in greatest dimensions. CT scan of the kidneys revealed slightly contrast-enhancing masses with irregular walls. Renal angiogram showed decreased blood supply to the areas coinciding with the masses consistent with renal scarring. There was no increased vascularity. This case demonstrates that renal scarring may masquerade as renal masses. A step-wise, comprehensive approach is necessary to rule out potentially malignant lesions in these patients. PMID- 11417755 TI - Psoas muscle abscess caused by Mycobacterium tuberculosis and Staphylococcus aureus: case report and review. AB - Tuberculosis psoas muscle abscess is currently an uncommon clinical entity in industrialized countries. It was considered the predominant cause of these abscesses in the early part of the 20th century as a result of complicated Potts disease. We describe the case of a psoas abscess caused by the combination of Mycobacterium tuberculosis and Staphylococcus aureus. In this patient, the abscess was not associated with Potts disease but with osteomyelitis of the iliac crest, in which the evaluation with magnetic resonance imaging was very specific in determining the extent and regional spread. PMID- 11417756 TI - Biomechanics of abdominal injuries. AB - Although considerable efforts have been advanced to investigate the biomechanical aspects of abdominal injuries, reviews have been very limited. The purpose of this article is to present a comprehensive review of the topic. Traumatic abdominal injuries occur due to penetrating or blunt loading. However, the present review is focused on blunt trauma. Because of the complexity of the abdomen, biomechanically relevant anatomical characteristics of the various abdominal organs are presented. The proposed mechanism of injury for these organs and methods for abdominal injury quantification are described. This is followed by a detailed analysis of the biomechanical literature with particular emphasis on experiments aimed to duplicate real world injuries and attempt to quantify trauma in terms of parameters such as force, deflection, viscous criteria, pressure criteria, and correlation of these variables with the severity of abdominal injury. Experimental studies include tests using primates, pigs, rats, beagles, and human cadavers. The effects of velocity, compression, padding, and impactor characteristics on tolerance; effects of pressurization and postmortem characteristics on abdominal injury; deduction of abdominal response corridors; and force-deflection responses (of the different abdominal regions and organs) are discussed. Output of initial research is presented on the development of a device to record the biomechanical parameters in an anthropomorphic test dummy during impact. Based on these studies and the current need for abdominal protection, recommendations are given for further research. PMID- 11417757 TI - Mechanics of endothelial cell architecture and vascular permeability. AB - Blood vessel walls form a selective barrier to the transport of materials between blood and tissue, and the endothelium contributes significantly to this barrier function. The role of the endothelium is particularly important in thin-walled vessels, such as venules, because during tissue inflammation the endothelial junctions widen in localized areas and gaps form, thus compromising the barrier function. The mechanisms of endothelial gap formation are still under question. In this review we describe what is known about the structure of endothelial cell cell junctions and how this structure can change during inflammation. We then consider two possible mechanisms by which endothelial gaps are formed: active endothelial cell contraction or breakdown of the junctional complex, followed by passive recoil. Using measured values of the mechanical properties of endothelial cells, and the forces to which they are subjected, we calculate that gap formation by breakdown of cellular adhesion, followed by passive recoil, is a feasible mechanism. Finally, since endothelial cell surfaces, including junctions, are coated with a glycocalyx, we consider the question of whether changes in the glycocalyx can markedly increase endothelial permeability. We conclude that gap formation can occur by active contraction or by breakdown of adhesion, depending on the inflammatory mediator, and that the responses of the glycocalyx may also play an important role in the regulation of microvascular permeability. PMID- 11417758 TI - Measuring the effectiveness of antiretroviral agents. AB - Considerable progress has been made recently in developing effective antiretroviral combination therapy that can suppress viral replication and delay disease progression in individuals infected with HIV. A range of up to 15 approved antiretroviral agents is now available, which target two different viral enzymes, while several agents are in clinical development. The rapid development and approval of antiretroviral agents, driven by the urgency of clinical need as well as the complexity of possible combinations, has precluded the extensive comparative clinical testing of regimens, which is necessary to establish the relative efficacy of various different agents. The lack of an appropriate animal model for HIV disease also increases reliance on in vitro measures. Several different in vitro and in vivo parameters have been defined in an attempt to quantify the effectiveness of antiretroviral agents, most importantly the 50% inhibitory and effective concentrations (IC50 and EC50). However, the clinical relevance of these measures is uncertain. Additionally, considerable variation exists in the usage of the terms 'IC50' and 'EC50' in recent publications in the literature. These issues pose interpretation problems to clinicians seeking information on the relative clinical efficacy of the agents. In this brief review, we attempt to clarify the different measures available and their potential utility for clinical decision-making, focusing particularly on the example of HIV protease inhibitors. There are many different quantifiable parameters that give information regarding the effectiveness of an antiviral drug. These include: inhibition of the viral target enzyme (inhibition constant, Ki); selectivity for viral versus host enzymes; inhibition of viral replication in cell culture (IC50); ratio of efficacy to cytotoxicity in vitro (therapeutic index); inhibition of viral replication or symptoms in an appropriate animal model of the disease (EC50); and the effect on surrogate markers, such as viral load or CD4 cell count, after administration to humans (in vivo EC50). Each of these different parameters gives valid information about the properties of an antiretroviral agent, which can help to build up a picture of its potential clinical utility relative to other drugs. However, to gain meaningful results, it is important to apply this information intelligently, understanding the limitations of each parameter. PMID- 11417759 TI - Laboratory guidelines for the practical use of HIV drug resistance tests in patient follow-up. AB - HIV drug resistance is one of the major limitations in the successful treatment of HIV-infected patients using currently available antiretroviral combination therapies. When appropriate, drug susceptibility profiles should be taken into consideration in the choice of a specific combination therapy. Guidelines recommending resistance testing in certain circumstances have been issued. Many clinicians have access to resistance testing and will increasingly use these results in their treatment decisions. In this document, we comment on the different methods available, and the relevant issues relating to the clinical application of these tests. Specifically, the following recommendations can be made: (i) genotypic and phenotypic HIV-1 drug resistance analyses can yield complementary information for the clinician. However, insufficient information currently exists as to which approach is preferable in any particular clinical setting; (ii) when HIV-1 drug resistance testing is required, it is recommended that testing be performed on plasma samples obtained before starting, stopping or changing therapy, on samples that have a viral load above the detection limit of the resistance test; (iii) the panel recommends that genotypic and phenotypic HIV 1 drug resistance testing for clinical purposes be performed in a certified laboratory under strict quality control and quality assurance standards; and (iv) the panel recommends that resistance testing laboratories provide clinicians with resistance reports that include a list of drug-related resistance mutations (genotype) and/or a list of drug-related fold resistance values (phenotype), with interpretations of each by an experienced virologist. The interpretation of genotypic and phenotypic analysis is a complex and developing science, and in order to understand HIV-1 drug resistance reports, communication between the requesting clinician and the expert that interpreted the resistance report is recommended. PMID- 11417760 TI - Virological response in multidrug-experienced HIV-1-infected subjects failing highly active combination regimens after shifting from lamivudine to didanosine. AB - The aim of this study (the RESCUE trial) was to verify the effect of a shift from a lamivudine-containing to a didanosine-containing regimen on viral replication in HIV-1-infected subjects who had experienced prior treatment failure. Sixteen patients (didanosine-experienced in 14/16 cases) were consecutively enrolled: eight patients shifted from lamivudine to didanosine without other changes in their drug regimen. The other eight shifted from lamivudine to didanosine and changed one or more of their other drugs according to their physician's judgement. At the time of the regimen shift, all the subjects exhibited a high level phenotypic resistance to both zidovudine and lamivudine with changes at codons 70-219 in 100% of cases, at codon 215 in 13 of 16 patients, and the M184V substitution in 13/16 patients. Phenotypic susceptibility to didanosine was maintained in the majority of cases (14/16) despite the high prevalence of changes at codon 184. A statistically significant decrease in viral load (P<0.005) without a significant increase in CD4 lymphocytes (P=0.514) was observed after 3 and 6 months from the introduction of the didanosine-containing regimen. These findings suggest the possibility of achieving a viral load response to didanosine-containing regimens in patients with reverse transcriptase (RT) M184V mutations who were previously treated with this drug and its possible use in salvage combinations. PMID- 11417761 TI - Predictors of virological response in HIV-infected patients to salvage antiretroviral therapy that includes nelfinavir. AB - Different salvage strategies have been used to regain control in patients with HIV who have virological failure on combination antiretroviral therapy. We conducted a cohort study of 63 extensively antiretroviral pretreated patients who initiated nelfinavir as part of salvage therapy, to determine predictors of virological response. The maximum HIV RNA response was >0.5 log10 copies/ml reduction in 43 patients (68%), including 21 patients (33%) who had suppression to <500 copies/ml. Corresponding response rates at 24 weeks were 41 and 19%, respectively. Responders and non-responders could not be distinguished by mean baseline HIV RNA or CD4 cell count, duration of prior protease inhibitor (PI) use, introduction of an initial non-nucleoside reverse transcriptase inhibitor or the number of antiretroviral agents changed when nelfinavir was added, likely reflecting the homogeneity of the population studied. The only parameter predictive of response was virus genotype. Response rates were lower in patients with increasing numbers of primary (P=0.045) or secondary (P=0.001) PI mutations. The addition of increasing numbers of reverse transcriptase mutations further impaired response rates (P=0.004). PMID- 11417762 TI - Durable HIV-1 suppression with indinavir after failing lamivudine-containing double nucleoside therapy: a randomized controlled trial. AB - OBJECTIVE: To assess the durability of the antiretroviral effect in plasma and cerebrospinal fluid (CSF) of antiviral therapy intensification, produced by the addition of indinavir from week 12 onwards to the original regimen of zidovudine/lamivudine or stavudine/lamivudine, after 72 weeks of follow-up using an ultrasensitive HIV-1 RNA assay. To assess CSF concentrations of indinavir at week 48. DESIGN: In a prospectively, randomized, open, single-centre study, antiretroviral-naive patients (CD4 cell count > or =200 cells/microl and a plasma HIV-1 RNA level 10,000 copies/ml) were assigned to a combination of zidovudine/lamivudine or stavudine/lamivudine. Indinavir could be added to the double nucleoside analogue regimen from week 12 or thereafter in case the plasma HIV RNA level was insufficiently suppressed (>500 copies/ml). RESULTS: Forty seven patients were enrolled (23 stavudine/lamivudine and 24 zidovudine/lamivudine), of whom 33 completed a follow-up of 72 weeks. Indinavir was added in 89% (42/47) of the patients. Only one discontinuation occurred due to virological failure. At week 72, the median plasma HIV-1 RNA levels in the zidovudine/lamivudine group had decreased from 4.80 log10 copies/ml to <500 copies/ml in 100% of patients and <50 copies/ml in 86.6% of the patients. In the stavudine/lamivudine group the plasma HIV-1 RNA decreased from 4.98 log10 copies/ml at baseline to <500 copies/ml in 100% of patients and <50 copies/ml in 66.7% of the patients. On an intent-to-treat basis these figures were 54.2 and 52.2% for zidovudine/lamivudine and stavudine/lamivudine, respectively, for the 50 copies/ml assay. The median CD4 cell count increased from 315 cells/microl, with 150 cells/microl in the zidovudine/lamivudine arm, and from 290 cells/microl, with 310 cells/microl in the stavudine/lamivudine arm (P=0.0001). However, the percentage of CD4 cells did not differ in each group. In the zidovudine/lamivudine group 9/10 and 5/5, and in the stavudine/lamivudine group 11/11 and 6/6 had a CSF HIV-1 RNA level <50 copies/ml at week 12 and 48, respectively. The CSF indinavir concentration ranged from 50 to 170 ng/ml. CONCLUSION: The long-term HIV-1 suppression observed in this study is remarkable, as adding a single antiretroviral agent to a failing regimen goes against current notions of adequate therapy. PMID- 11417763 TI - Prevalence of genotypic resistance among antiretroviral drug-naive HIV-1-infected patients in Belgium. AB - OBJECTIVES: To estimate the prevalence and the evolution over time (1995-1998) of genotypic resistance to antiviral drugs in antiretroviral drug-naive HIV-1 infected patients in Belgium. DESIGN: Belgian Aids Reference Laboratories provided retrospective samples and clinical data from antiretroviral drug-naive HIV-1-infected patients who visited the hospital for the first time in 1995 (n=45), 1997 (n=75) and 1998 (n=111). Genotypic resistance to the three available classes of drugs was monitored using the Line Probe Assay (Innogenetics, Gent, Belgium). Additionally, ARMS-151 was performed for scoring multinucleoside resistance. RESULTS: The prevalence of genotypic resistance at baseline to nucleoside analogue reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs) were each between 10% and 20% for 1995, 1997 and 1998 without an increasing trend over time. For NRTIs, resistance mutations were mainly related to zidovudine in 1995, whereas in 1997 and 1998 baseline resistance was scored for zidovudine, lamivudine or for both drugs simultaneously. No patients displayed the multi-nucleoside resistance Q151M mutation. Baseline resistance mutations to protease inhibitors (PIs) did not rise significantly: 4.4% in 1995, 8% in 1997 and 9.9% in 1998. When scoring any resistance-related mutation, 26.6% displayed genotypic baseline resistance in 1995, 26.6% in 1997 and 31.5% in 1998. DISCUSSION: The prevalence of genotypic baseline resistance to any drug, as scored with LiPA, in naive HIV-1 patients in Belgium is 29%, with baseline resistance mutations to one or several drugs from all available classes of antiviral drugs. The ability of LiPA to pick up minor variants could be an explanation for the higher overall prevalence we observe, when compared to recent estimates in other countries of 16.3% and 22%, which were based on sequencing methods. According to the European guidelines for resistance testing, resistance testing in Belgium before starting antiviral therapy should be considered. PMID- 11417764 TI - Resistance profiles to antiretroviral drugs in HIV-1 drug-naive patients in Argentina. AB - The drug resistance profile of treatment-naive HIV-infected individuals living in Buenos Aires, Argentina, was studied. Samples taken from 94 drug-naive individuals with established HIV infection and 13 patients with primary HIV infection were assessed by nucleotide sequencing and LIPA. The prevalence of drug associated primary mutations in individuals with established infection was very low. In the viral protease region, 1/86 (1.2%) individuals carried the D30N mutation, whereas 1/85 (1.2%) had the M41L mutation in the reverse transcriptase (RT) region. Secondary mutations in both the protease and RT regions were found in almost 90% of the individuals. In individuals with primary infection, primary mutations were detected in 2/13 (15.4%) patients, one of them carrying M461 mutation in the protease while the other patient had a mutation at codon 184 of the RT. In accordance with current drug resistance testing guidelines, the results of this study suggest that susceptibility tests need not be performed at this time prior to initiation of antiretroviral therapy in HIV-1-infected people in Argentina. However, the public health implications of this subject warrant follow-up studies that will examine a larger number of drug-naive patients, not only in Buenos Aires but also in other major Argentinian cities and in rural areas. PMID- 11417765 TI - Cervicovaginal HIV-1 shedding in pregnant women near delivery. PMID- 11417766 TI - Antiretroviral therapy and the lipodystrophy syndrome. AB - 'Lipodystrophy syndrome' in the setting of HIV infection has come to encompass a collection of morphological and metabolic abnormalities linked with the use of antiretroviral therapy and other risk factors. We review the clinical literature on this subject as it has evolved historically, taking pertinent methodological issues into account. PMID- 11417767 TI - PTSD after spontaneous abortion. PMID- 11417768 TI - PTSD after spontaneous abortion. PMID- 11417769 TI - Helping physicians make evidence-based decisions. PMID- 11417770 TI - Physicians need practical tools to treat the complex problems of overweight and obesity. PMID- 11417771 TI - Obesity: assessment and management in primary care. AB - Obesity is a complex, multifactorial condition in which excess body fat may put a person at health risk. National data indicate that the prevalence of obesity in the United States is increasing in children and adults. Reversing these trends requires changes in individual behavior and the elimination of societal barriers to healthy lifestyle choices. Basic treatment of overweight and obese patients requires a comprehensive approach involving diet and nutrition, regular physical activity, and behavioral change, with an emphasis on long-term weight management rather than short-term extreme weight reduction. Physicians and other health professionals have an important role in promoting preventive measures and encouraging positive lifestyle behaviors, as well as identifying and treating obesity-related comorbidities. Health professionals also have a role in counseling patients about safe and effective weight loss and weight maintenance programs. Recent evidence-based guidelines from the National Heart, Lung, and Blood Institute, as well as recommendations from the American Academy of Pediatrics, American Association of Clinical Endocrinologists/American College of Endocrinology, American Obesity Association, U.S. Clinical Preventive Services Task Force, Institute of Medicine, and World Health Organization can be consulted for information and guidance on the identification and management of overweight and obese patients. PMID- 11417772 TI - Valvular heart disease: review and update. AB - People with valvular heart disease are living longer, with less morbidity, than ever before. Advances in surgical techniques and a better understanding of timing for surgical intervention account for increased rates of survival. Echocardiography remains the gold standard for diagnosis and periodic assessment of patients with valvular heart disease. Generally, patients with stenotic valvular lesions can be monitored clinically until symptoms appear. In contrast, patients with regurgitant valvular lesions require careful echocardiographic monitoring for left ventricular function and may require surgery even if no symptoms are present. Aside from antibiotic prophylaxis, very little medical therapy is available for patients with valvular heart disease; surgery is the treatment for most symptomatic lesions or for lesions causing left ventricular dysfunction even in the absence of symptoms. PMID- 11417773 TI - Neurologic complications of scuba diving. AB - Recreational scuba diving has become a popular sport in the United States, with almost 9 million certified divers. When severe diving injury occurs, the nervous system is frequently involved. In dive-related barotrauma, compressed or expanding gas within the ears, sinuses and lungs causes various forms of neurologic injury. Otic barotrauma often induces pain, vertigo and hearing loss. In pulmonary barotrauma of ascent, lung damage can precipitate arterial gas embolism, causing blockage of cerebral blood vessels and alterations of consciousness, seizures and focal neurologic deficits. In patients with decompression sickness, the vestibular system, spinal cord and brain are affected by the formation of nitrogen bubbles. Common signs and symptoms include vertigo, thoracic myelopathy with leg weakness, confusion, headache and hemiparesis. Other diving-related neurologic complications include headache and oxygen toxicity. PMID- 11417774 TI - Information from your family doctor. Medical problems of recreational scuba diving. PMID- 11417775 TI - Fishhook removal. AB - Fishing is a common recreational sport. While serious injuries are uncommon, penetrating tissue trauma involving fishhooks frequently occurs. Most of these injuries are minor and can be treated in the office without difficulty. All fishhook injuries require careful evaluation of surrounding tissue before attempting removal. Ocular involvement should prompt immediate referral to an ophthalmologist. The four most common techniques of fishhook removal and injury management are described in this article. The choice of the method for fishhook removal depends on the type of fishhook embedded, the location of the injury and the depth of tissue penetration. Occasionally, more than one removal technique may be required for removal of the fishhook. The retrograde technique is the simplest but least successful removal method, while the traditional advance and cut method is most effective for removing fishhooks that are embedded close to the skin surface. The advance and cut technique is almost always successful, even for removal of large fishhooks. The string-yank method can be used in the field and can often be performed without anesthesia. Wound care following successful removal involves extraction of foreign bodies from the wound and the application of a simple dressing. Prophylactic antibiotics are generally not indicated. Tetanus status should be assessed and toxoid administered if needed. PMID- 11417776 TI - Evaluation and management of the AGUS Papanicolaou smear. AB - Atypical glandular cells on Papanicolaou smears are an unusual but important cytologic diagnosis. The Bethesda system classifies atypical glandular cells of undetermined significance (AGUS) as glandular cells that demonstrate nuclear atypia appearing to exceed reactive or reparative changes but lacking unequivocal features of adenocarcinoma. AGUS occurs in approximately 0.18 to 0.74 percent of all cervical smears. Because of the high likelihood that AGUS is associated with significant clinical disease, simply repeating the Papanicolaou smear is not sufficient for the management of AGUS. Unlike women with atypical squamous cells of undetermined significance, a significant percentage of women with AGUS will have more serious lesions, such as high-grade preinvasive squamous disease, adenocarcinoma in situ, adenocarcinoma or invasive cancers from sites other than the cervix. Colposcopic examination is recommended for all women with a cytologic diagnosis of AGUS. Those women with AGUS that is suspicious for adenocarcinoma should undergo cervical conization, even in the absence of detectable abnormalities on colposcopic examination. PMID- 11417777 TI - Should we recommend nicotine replacement therapy? PMID- 11417778 TI - Information from your family doctor. Using the nicotine patch, nicotine gum, nicotine nasal spray or nicotine inhaler. PMID- 11417779 TI - Photo quiz. Sore throat and difficulty breathing. PMID- 11417780 TI - ACOG practice bulletin on preventing deep venous thrombosis and pulmonary embolism. . PMID- 11417781 TI - Dealing with adolescent Latino patients. PMID- 11417782 TI - The domain specificity and generality of mental contamination: accuracy and projection in judgments of mental content. AB - In this study we examined individual differences across two tasks requiring people to judge the mental contents of the minds of other people--an opinion prediction task and a knowledge prediction task. The tendency to overproject one's own mental contents in both of these tasks has been interpreted as an instance of mental contamination by Wilson and Brekke (1994). Results demonstrate no domain generality in the process of projecting one's own internal states onto predictions about the internal states of others. Furthermore, projection was efficacious in the opinion prediction task but not in the knowledge prediction task. The differing consequences of mental contamination across these tasks was moderated by the presence of other diagnostic cues that were negatively correlated with the diagnosticity of one's own mental contents in the knowledge prediction task but not in the opinion prediction task. Mental contamination was largely unrelated to cognitive ability or to styles of epistemic regulation. However, predictive accuracy (and its primary determinant--use of other diagnostic cues) was correlated across the two tasks, and was also related to cognitive ability and styles of epistemic regulation. The results are interpreted within the context of two-process models of cognitive functioning (e.g. Evans & Over, 1996; Sloman, 1996). PMID- 11417783 TI - Priming the access to names of famous faces. AB - There is a continuing controversy in models of face identification concerning the level of access to names relative to semantic information. In order to determine whether names are accessed sequentially after or in parallel to semantic information, we studied participants' speeded decisions about famous faces that were primed by partial semantic or partial name information. Decisions that required the access to the celebrity's name (one or more forename syllables, Expt 1) were significantly primed by partial name primes (initials or name fragments). However, at variance with sequential stage models, no reliable priming was observed by partial semantic primes (information about nationality, occupation, or whether a person was dead or alive). Moreover, there was a clear and consistent priming effect by partial semantic primes if the task was a nationality (British or American) decision that required the access to semantic information (Expt 2), demonstrating the effectiveness of these primes. The effects of partial name primes on nationality decisions were less consistent, with a significant effect for name fragments but not initials. However, effects of name primes were generally greater for syllable decisions than nationality decisions, and effects of semantic primes were generally greater for nationality decisions than syllable decisions. Taken together, these results favour a model of parallel rather than sequential access and suggest some degree of independence in the access to personal semantics and names. PMID- 11417784 TI - Effects of spatial and nonspatial cognitive activity on postural stability. AB - Is postural stability controlled automatically, or is it affected by concurrent cognitive activity? Are the effects influenced by the nature of the cognitive activity required, and do they increase in old age? To address these questions, 70 participants aged 20-79 years were asked to stand as still as possible on a force platform (postural control task) while performing (a) no cognitive task, (b) a spatial memory task, and (c) a nonspatial memory task. The memory tasks were also performed while seated as a comparison condition. Both spatial and nonspatial memory recall declined with increasing age but were unaffected by position (standing vs. seated). Postural stability declined with age; moreover, there was support for an earlier finding that age decline was greatest when performing the spatial memory task. Each recording period was split into two phases which, for the spatial and nonspatial memory tasks, corresponded to encoding and maintaining the stimuli. In comparison with no task, participants were more stable when encoding stimuli (particularly in the spatial task), but they were less stable when maintaining stimuli (particularly in the nonspatial task). The results suggest that postural stability can be affected by cognitive activity in complex ways, depending on the age of participants, the type of cognitive task (spatial vs. nonspatial), and the cognitive processing required (encoding vs. maintenance). PMID- 11417785 TI - Bisensory augmentation: a speechreading advantage when speech is clearly audible and intact. AB - Reisberg, McLean, and Goldfield (1987) have shown that vision plays a part in the perception of speech even when the auditory signal is clearly audible and intact. Using an alternative method the present study replicated their finding. Clearly audible spoken messages were presented in audio-only and audio-visual conditions, and the adult participants' resulting comprehension was measured. Stories were presented in French (Expt 1), in a Glaswegian accent (Expt 2), and by presenting spoken information that was semantically and syntactically complex (Experiment 3). Three separate groups of 16 adult female participants aged 19-21 participated in the three experiments. In all three experiments, comprehension improved significantly when the speaker's face was visible. PMID- 11417786 TI - The nature of attentional differences between groups of children differentiated by teacher ratings of attention and hyperactivity. AB - Two groups of children, one with good attention and low hyperactivity and the other with poor attention and high hyperactivity, were formed on the basis of teachers' ratings. The groups were tested on a large number of tests of attentional performance, covering selective attention, divided attention, sustained attention and executive function. Analysis of group differences, with verbal and non-verbal intelligence controlled statistically, indicated that the groups demonstrated significant differences on a variety of measures. Principal components analysis of the tests of attention isolated two components, one of which significantly distinguished the two groups. It is suggested that this component reflected aspects of executive control of attention. The measures which discriminated the groups most clearly were efficiency of visual search involving two targets in alternation, the number of correct responses on a specially devised version of the Wisconsin Card Sorting Task and a task measuring the ability to inhibit a dominant response. It is suggested that all these tasks required a combination of selective attention and executive control. It was also noted that the poor attention group produced bigger individual differences on all measures, indicating that this group may include a variety of weaknesses which require more precise analysis to distinguish them. PMID- 11417787 TI - People's insight into their own behaviour and speech content while lying. AB - The present experiment examined participants' insight into their own behaviour and speech content while lying. It was hypothesized that participants would believe that while lying they show more behaviour stereotypical of lying than they in fact do (Hypothesis 1), whereas they would believe that their own speech content while lying contains fewer stereotypical features than in fact is true (Hypothesis 2). A stereotypical response was defined as a response people generally believe liars usually show. A total of 86 nursing students were interviewed twice about a film they had just seen. During one interview they were asked to tell the truth whereas they had to lie in the other interview. All interviews were videotaped, transcribed and then scored by independent coders. The coders' analyses reveal participants' actual behaviour and speech content. Participants themselves were asked to indicate in a questionnaire how they believed they behaved and what they believed they said in both interviews. To test the hypotheses, comparisons were made between participants' actual responses and their beliefs about their own responses. The results support both hypotheses and implications of these outcomes for the detection of deception are discussed. PMID- 11417788 TI - Female and male perceptions of female physical attractiveness in front-view and profile. AB - Two important cues to female physical attractiveness are body mass index (BMI) and shape. In front view, it seems that BMI may be more important than shape; however, is it true in profile where shape cues may be stronger? There is also the question of whether men and women have the same perception of female physical attractiveness. Some studies have suggested that they do not, but this runs contrary to mate selection theory. This predicts that women will have the same perception of female attractiveness as men do. This allows them to judge their own relative value, with respect to their peer group, and match this value with the value of a prospective mate. To clarify these issues we asked 40 male and 40 female undergraduates to rate a set of pictures of real women (50 in front-view and 50 in profile) for attractiveness. BMI was the primary predictor of attractiveness in both front and profile, and the putative visual cues to BMI showed a higher degree of view-invariance than shape cues such as the waist-hip ratio (WHR). Consistent with mate selection theory, there were no significant differences in the rating of attractiveness by male and female raters. PMID- 11417790 TI - Essay review: Making shell-shock. PMID- 11417791 TI - Identification and localization of connexin26 within the photoreceptor-horizontal cell synaptic complex. AB - Connexin26 (Cx26) is a member of the family of integral membrane proteins that normally form intercellular gap junctional channels. We have used Western blotting, immunofluorescence, immunoelectron microscopy, and single-cell reverse transcriptase polymerase chain reaction amplification (RT-PCR) to analyze the expression and cellular localization of Cx26 in the carp retina. In the outer plexiform layer, strong clustered Cx26 immunolabeling was concentrated at and restricted to the terminal dendrites of horizontal cells. Single-cell RT-PCR confirmed the expression of Cx26 in carp retinal horizontal cells. 248-bp fragments amplified from cDNAs of four different horizontal cells were cloned and each nucleotide sequence encodes a protein fragment (AA 104-185) with highly significant homology to rat and mouse Cx26. Immunoelectron microscopy revealed that only the invaginating dendrites of horizontal cells in intimate lateral association with the presynaptic ribbon complex were labeled. No labeling was found at the photoreceptor membrane and there was no septalaminar structure, indicative of gap junctions, between photoreceptors and horizontal cells. The focal location of Cx26 at the membrane of the dendritic tips of horizontal cells and the lack of gap junctional morphology suggests that Cx26 might form hemichannels. PMID- 11417792 TI - Evidence for a visual subsector within the zona incerta. AB - Here we examine the patterns of connections between the zona incerta (ZI) of the thalamus and the major visual centers of the rat brain, namely the retina, dorsal lateral geniculate nucleus (LGd), superficial layers of the superior colliculus (SCs), and occipital cortex (Ocl). Injections of the tracers biotinylated dextran or cholera toxin subunit b were made into each of these centers, as well as ZI itself, by using stereotaxic coordinates. Rat brains were then aldehyde-fixed and processed using standard methods. We show that the retina, LGd, SCs, and Ocl all have connections with ZI; moreover, that each of these connections make a very distinct territory or subsector within the most lateral ZI regions. This subsector of connectivity with the visual centers does not respect the well defined cytoarchitectonic sectors of ZI, being made up of small zones in the dorsal, ventral, and caudal sectors. Often, a distinctive "horse-shoe" pattern is evident, particularly after retinal and Ocl injections. Tracer injections into topographically distinct regions of the LGd. SCs, or Ocl results in no shift in the spatial location of labelling within ZI; after each injection, labelling is always seen within the lateral edge of the nucleus. Labelled terminals and cells are seen after LGd and SCs injections, while only labelled terminals are seen after retinal and Ocl injections. Although the precise function of this novel visual subsector is not known, these early findings suggest that ZI may be in a position to integrate visual information together with the other somatosensory, motor, and visceral information that it receives. PMID- 11417793 TI - Contributions of AMPA- and kainate-sensitive receptors to the photopic electroretinogram of the Xenopus retina. AB - The effects of kainate receptor-preferring glutamate ligands were tested on the electroretinogram (ERG) of the Xenopus retina. Kainate, domoic acid, and 5 iodowillardiine (20-100 microM) acted similarly in every respect. They increased peak amplitudes of the ERG a-, b-, and d-waves significantly over controls. The AMPA-specific antagonist, GYKI 53655, prevented a kainate-induced increase in ERG a- and d-waves, but was without effect on an increase in the b-wave. Once the effect of agonist on the b-wave had peaked, the ERG began to subside, leading to its nearly complete disappearance within 20 min. Prior exposure to GYKI followed by a combination of GYKI + agonist did not significantly slow the rate of b-wave disappearance. Our results indicate that (1) AMPA receptors contribute to ERG a- and d-waves. (2) The kainate-evoked increase in ERG a-, b-, and d-waves probably results, in part, from an excitotoxic swelling of inner retinal processes. (3) The inner retina has a population of GYKI-resistant, kainate-sensitive receptors which may contribute to b-wave generation. PMID- 11417794 TI - Sequential recovery of sensitivity to negative and positive contrasts during optic nerve regeneration in goldfish. AB - A psychophysical procedure, classical conditioning of respiration, was used to measure contrast sensitivity to positive- and negative-contrast discs (8-deg diameter) in goldfish after crushing one optic nerve intraorbitally. In five out of six fish, sensitivity recovered to normal. Recovery times for negative contrasts were significantly shorter than for positive contrasts. The average times postcrush of initial responding to negative and positive contrasts were 23.8 and 30.6 days, and for threshold to come within 0.5 log unit of control values was 29.8 and 39.8 days, respectively. Thereafter, recovery to normal sensitivity was significantly faster for positive contrasts. These results parallel prior observations of neural activity in tectum after optic nerve crush: an early phase of OFF responding followed by a more sudden recovery of ON responding. PMID- 11417795 TI - Neural and perceptual adjustments to dim light. AB - At reduced luminance levels, the visual system integrates light over extended periods of time. Although the general effects of this process are known, specific changes in the visual cortex have not been identified. We have studied the physiological changes that occur during a transition from high to low luminance by measurements of single neurons in the cat's primary visual cortex. Under low luminance conditions, we find increased latencies, expanded temporal responses, and a loss of temporal structure. This results in temporal-frequency tuning curves that are peaked at relatively low frequencies. To examine parallel perceptual changes, we compared perceived temporal frequency in human subjects under high- and low-luminance conditions. Low-luminance flickering patterns are perceived to modulate at relatively high rates. This occurs even though peak sensitivity is shifted to relatively low temporal frequencies. To explore further the perceptual component, we measured perceived temporal frequency in human subjects with unilateral optic neuritis for whom optic nerve transmission is known to be relatively slow and generally similar to the normal physiological state under low luminance. These subjects also perceive relatively high modulation rates through their affected eye. Considered together, these results demonstrate an inverse relationship between the physiological and the perceptual consequences of reduced stimulus luminance. This relationship may be accounted for by shifts of neuronal population responses between high- and low-luminance levels. PMID- 11417796 TI - Long-range interactions in the lateral geniculate nucleus of the New-World monkey, Callithrix jacchus. AB - Visual stimulation of zones extending beyond the classical receptive field can modulate the contrast gain of neurons in the lateral geniculate nucleus (LGN) of cats, but little is known about the effect of extra-classical visual stimulation on the LGN of primates. Hence, we compare the effect of long-range interactions in parvocellular and magnocellular LGN layers of the marmoset monkey Callithrix jacchus using optimal, incremental spots flashed on the classical receptive field either alone or simultaneously with the shift of a grating (98% contrast; 0.1 cycles/deg) confined to a peripheral annulus (radii: 5-15 deg). The contrast required to drive the response halfway to saturation (c50) of most LGN neurons was raised by remote pattern shifts. The c50 ratio [(shift+spot)/spot] in OFF center magnocellular neurons was significantly higher than in OFF-center parvocellular neurons. OFF-center magnocellular neurons closer to the fovea (<10 deg eccentricity) tended to have a higher c50 ratio than in more peripheral neurons. A significant drop in visual sensitivity to 25% contrast spots was observed during remote motion: d' fell from 1.8 to 1.4 in parvocellular neurons and from 2.2 to 1.7 in magnocellular neurons. Such long-range interactions produce a reduction in visual sensitivity by changing the gain of the geniculate relay and point to an inhibitory, motion-sensitive extra-classical receptive field in both parvocellular and magnocellular pathways, which may be involved in saccadic suppression and attentional mechanisms in early vision. PMID- 11417797 TI - Dynamic properties of retino-geniculate synapses in the cat. AB - Simultaneous recordings from relay cells in the lateral geniculate nucleus (LGN) and their retinal afferents were used to examine the rules governing the transmission of spikes across the retino-geniculate synapse. Retinal spikes that terminate short retinal interspike intervals are much more likely to be transmitted across the synapse than spikes terminating longer intervals. This facilitation can be observed for interspike intervals as long as 50 ms when retinal firing rates are low, but the range of effective intervals decreases exponentially as retinal firing rate increases. Contribution, the fraction of LGN spikes triggered by an individual retinal afferent, is typically much higher during visual stimulation than during maintained activity, and these differences are unrelated to presynaptic or postsynaptic firing rate. It is suggested that this effect is a manifestation of increased synchronization of spikes among retinal afferents to the geniculate cell during structured visual stimulation, and that this synchronization offers a means of enhancing signal-to-noise ratio at the retino-geniculate synapse. Cross-correlograms between geniculate burst spikes and retinal afferents often contain two distinct peaks; a short latency peak that results from direct coupling between burst spikes and retinal input spikes, and a longer latency peak that represents indirect coupling in which retinal spikes trigger the calcium spike underlying the burst. Direct coupling is most likely to occur for the later spikes in the burst, and is present regardless of whether the calcium spike underlying the burst is triggered by the same or a different retinal afferent. These results further illuminate the relationship between tonic and burst modes of retino-geniculate transmission and indicate that bursts in LGN relay cells can be viewed as a mechanism of signal amplification, producing signals whose timing is potentially related to the temporal structure of a stimulus, independent of presynaptic and postsynaptic firing rate. This mechanism also appears to capitalize on the synchronization that is present among parallel retinal afferents to a geniculate cell. PMID- 11417798 TI - Organization of the inner retina following early elimination of the retinal ganglion cell population: effects on cell numbers and stratification patterns. AB - The present study has examined the effects of early ganglion cell elimination upon the organization of the inner retina in the ferret. The population of retinal ganglion cells was removed by optic nerve transection on the second postnatal day, and retinas were subsequently studied in adulthood. Numbers of amacrine and bipolar cells were compared in the nerve-transected and nerve-intact retinas of operated ferrets, while stratification patterns within the inner plexiform layer were compared in these and in normal ferret retinas. Early ganglion cell elimination was found to produce a 25% reduction in the population of glycine transporter-immunoreactive amacrine cells, and 18 and 15% reductions in the populations of parvalbumin and calbindin-immunoreactive amacrine cells, respectively. GABAergic amacrine cells were also reduced by 34%. The number of calbindin-immunoreactive displaced amacrine cells, by contrast, had increased in the ganglion cell-depleted retina, being three times their normal number. Other amacrine and bipolar cell types were unaffected. Despite these changes, the stratification patterns associated with these cell types remained largely intact within the inner plexiform layer. The present results demonstrate a class specific dependency of inner retinal neurons upon the ganglion cell population in early postnatal life, but the ganglion cells do not appear to provide any critical signals for stratification within the inner plexiform layer, at least not after birth. Since they themselves do not produce stratified dendritic arbors until well after birth, the signals for stratification of the bipolar and amacrine cell processes should arise from other sources. PMID- 11417799 TI - Rod photopigment deficits in albinos are specific to mammals and arise during retinal development. AB - Adult albino mammals have specific retinal defects, including reduced numbers of rod photoreceptors. To examine when this rod deficit arises and whether it exists in nonmammalian albinos, we have used absorbance spectrophotometry to measure photopigment levels in dark-adapted eyes taken from three groups of pigmented and albino animals: adult rodents (rats and mice), developing rats, and mature Xenopus frogs. Rhodopsin concentrations were consistently and significantly reduced in mammalian albinos compared to their wild-type counterparts from before the time of eye opening, but photopigment levels were similar in frogs of both pigmentation phenotypes. The results strongly suggest that deficits in the rod cell population arise early in development of the mammalian albino retina, but do not generalize to nonmammalian mutants lacking retinal melanin. PMID- 11417800 TI - The effects of saccadic eye movements on the activity of geniculate relay neurons in the monkey. AB - Saccadic suppression is the reduced visibility that occurs during saccadic eye movements. Recent psychophysical studies have suggested that this is due to a reduction in responsiveness of magnocellular (M), but not parvocellular (P), cells of the lateral geniculate nucleus. To address this and other phenomena of responsiveness during saccades, we recorded from geniculate neurons in the behaving monkey before, during, and after saccades. Specifically, we measured neuronal responses to a flashing, whole-field illumination. Contrary to the prediction, most M neurons showed pronounced enhancement of visual activity during saccades, whereas such responsiveness of parvocellular (P) neurons was not significantly affected by saccades. We also analyzed the extent to which saccades affected burst firing, which results from activation of a voltage-dependent Ca2+ conductance. We found that both M and P cells displayed a significant suppression of burst firing during saccades. These results do not support the idea that saccadic suppression has an obvious substrate in reduced responsiveness of geniculate cells, but this suppression may be related to an increased visual threshold for detection associated with reduced burst firing. PMID- 11417801 TI - Selective activation of visual cortex neurons by fixational eye movements: implications for neural coding. AB - During normal vision, when subjects attempt to fix their gaze on a small stimulus feature, small fixational eye movements persist. We have recorded the impulse activity of single neurons in primary visual cortex (V1) of macaque monkeys while their fixational eye movements moved the receptive-field activating region (AR) over and around a stationary stimulus. Three types of eye movement activation were found. (1) Saccade cells discharged when a fixational saccade moved the AR onto the stimulus, off the stimulus, or across the stimulus. (2) Position/drift cells discharged during the intersaccadic (drift) intervals and were not activated by saccades that swept the AR across the stimulus without remaining on it. To activate these neurons, it was essential that the AR be placed on the stimulus and many of these cells were selective for the sign of contrast. They had smaller ARs than the other cell types. (3) Mixed cells fired bursts of activity immediately following saccades and continued to fire at a lower rate during intersaccadic intervals. The tendency of each neuron to fire transient bursts or sustained trains of impulses following saccades was strongly correlated with the transiency of its response to stationary flashed stimuli. For one monkey, an extraretinal influence accompanying fixational saccades was identified. During natural viewing, the different eye movement classes probably make different contributions to visual processing. Position/drift neurons are well suited for coding spatial details of the visual scene because of their small AR size and their selectivity for sign of contrast and retinal position. However, saccade neurons transmit information that is ambiguous with respect to the spatial details of the retinal image because they are activated whether the AR lands on a stimulus contour, or the AR leaves or crosses the contour and lands in another location. Saccade neurons may be involved in constructing a stable world in spite of incessant retinal image motion, as well as in suppressing potentially confusing input associated with saccades. PMID- 11417802 TI - Voltage-activated Ca2+ channels and ionotropic GABA receptors localized at axon terminals of mammalian retinal bipolar cells. AB - A preparation of isolated presynaptic terminals of rat retinal rod bipolar cells was developed. Patch-clamp recordings were performed on the isolated terminal to determine the type(s) of voltage-activated Ca2+ channels and the contribution of GABA(A) and GABA(C) receptor-mediated currents localized in the terminal region. Both low-voltage-activated (LVA) and high-voltage-activated (HVA) Ca2+ currents, with properties similar to those found in intact cell recordings, were observed in the isolated terminal recordings. Consistent with previous studies, the HVA Ca2+ currents are L-type since the currents were blocked by low micromolar concentrations of nimodipine and potentiated by BayK 8644. Also, both GABA(A) and GABA(C) receptor-mediated currents were observed in the isolated terminal. The current density of GABA(C) receptors in the terminal was more than three times higher than that in the soma. In contrast, the current density of GABA(A) currents between the terminal and the soma was not significantly different. Assessed by 100 microM GABA, the contributions of GABA(A) and GABA(C) receptors to the total GABA-mediated currents at the terminal were comparable. This study directly demonstrates the localization of LVA Ca2+ channels at the axon terminal of mammalian rod bipolar cells, suggesting that LVA Ca2+ channels may play a role in bipolar cell transmitter release. Results of this study also support the notion that both types of ionotropic GABA receptors regulate synaptic transmission in mammalian rod bipolar cells. In addition, this study reports for the first time the feasibility of direct patch-clamp recordings of isolated axon terminals of mammalian retinal bipolar cells. The isolated presynaptic terminal preparation of mammalian retinal bipolar cells could be a valuable system for the study of transmitter release in the central nervous system (CNS). PMID- 11417803 TI - Evidence for spatial regularity among retinal ganglion cells that project to the accessory optic system in a frog, a reptile, a bird, and a mammal. AB - The vertebrate retina contains only five major neuronal classes but these embrace a great diversity of discrete types, many of them hard to define by classical methods. Consideration of their spatial distributions (mosaics) has allowed new types, including large ganglion cells, to be resolved across a wide range of vertebrates. However, one category of large ganglion cells has seemed refractory to mosaic analysis: those that project to the accessory optic system (AOS) and serve vestibulocerebellar mechanisms of motion detection and image stabilization. Whenever AOS-projecting cells have been analyzed by nearest-neighbor methods, their distribution has appeared almost random. This is puzzling, because most aspects of visual processing require the visual scene to be sampled regularly. Here, spatial correlogram methods are applied to distributions of large ganglion cells, labeled retrogradely from the AOS in frogs, turtles, and rats, and to the AOS-projecting displaced ganglion cells of chickens. These methods reveal hidden spatial order among AOS-projecting populations, of a form that can be simulated either by superimposing a single regular mosaic on a random population or, more interestingly, by overlapping three or more regular, similar but spatially independent mosaics. The rabbit is known to have direction-selective ganglion cells (not, however, AOS projecting) that can be subdivided into functionally distinct, regular mosaics by their tracer-coupling patterns even though they are morphologically homogeneous. The present results imply that the direction selective AOS-projecting ganglion cells of all vertebrates may, likewise, be subdivided into regular, independent mosaics. PMID- 11417804 TI - Functional and morphological assessment of age-related changes in the choroid and outer retina in pigeons. AB - We sought to determine if choroidal and outer retinal deterioration occur with age in pigeons, as they do in other species, and investigated the relationship between age-related retinal and choroidal changes. In 64 pigeons ranging in age over the pigeon lifespan (0.5-20 years), we measured some or all among the following parameters: choroidal blood flow (ChBF) by laser Doppler flowmetry, choroidal thickness and choriocapillary vessel abundance by LM histology, choriocapillary endothelial cell transport specializations by EM histology, acuity by behavioral methods, and degenerating photoreceptor abundance and total photoreceptor abundance by LM histology. Regression and Receiver Operator Curve (ROC) analyses were used to characterize the pattern of age-related changes and determine the ages at or by which significant changes occurred. For the 45 birds for which we measured choroidal parameters, choriocapillary vessel abundance showed a curvilinear decline with age and half of this decline occurred by 3.5 4.6 years. The endothelial cell transport specializations called channels also declined curvilinearly with age. Choroidal thickness was slightly increased between the ages of 3-6 years, and thereafter declined steadily so that choroidal thickness in the oldest birds was half that in the youngest. ChBF showed an abrupt decline of about 20% at 4 years and a further 20% decline thereafter. In the 53 birds for which we obtained visual acuity and/or photoreceptor data, we observed a curvilinear decline in acuity (with half the decline having occurred by 8 years) and a prominent stepwise decline of about 20% in photoreceptor abundance at 4.7 years, followed by further decline thereafter. The period of major photoreceptor loss coincided with ages during which about 10% of photoreceptors appeared to show degenerative changes (4-8 years of age). Using partial correlation analysis with the common effect of age held constant, ChBF was found to have a positive correlation with acuity. Our results show that ChBF and choroidal vascularity decline significantly with age in pigeons, as do acuity and photoreceptor abundance. Our statistical analyses suggest that prominent choroidal vascular decline preceded the visual decline, and that there is a positive relationship between choroidal and visual functions. Thus, our findings are consistent with the view that age-related decline in choroidal function might contribute to age-related vision loss in pigeons. PMID- 11417805 TI - A comparative morphometric analysis of the optic nerve in two cetacean species, the striped dolphin (Stenella coeruleoalba) and fin whale (Balaenoptera physalus). AB - A comparative study was made on one Mysticete (the fin whale, Balaenoptera physalus) and one Odontocete species (the striped dolphin, Stenella coeruleoalba) by measuring several morphological characteristics seen in cross sections of the optic nerve. We found that the two cetacean nerves share a number of specializations that distinguish them from the optic nerve of terrestrial mammals. Fiber density is approximately two-fold lower than in land mammals. A corresponding increase in the cross-sectional area occupied by astrocytes is observed. A population of "giant" (up to 15 microm in diameter) optic axons is present in both the B. physalus and the S. coeruleoalba nerve. It is argued that these features probably reflect common adaptations to the constraints imposed by the aquatic environment. "Giant" optic axons might ensure short-latency detection of preys and other targets during navigation while the increased astroglial content might be related to the maintenance of neuronal function during periods of anaerobic metabolism under water. PMID- 11417806 TI - Dopamine depletion with 6-OHDA enhances dopamine D1-receptor modulation of potassium currents in retinal bipolar cells. AB - Ascorbate modulates IK(V) of ON-type mixed rod/cone bipolar cells (Mb) in the goldfish retinal slice through a dopamine D1/G-protein/PKA-coupled mechanism. We investigated the effects of dopamine depletion with intraocular injections of 6 OHDA on IK(V) and its modulation by ascorbate over 1-7 weeks following 6-OHDA treatment. Dopamine depletion was verified by tyrosine hydroxylase immunocytochemistry. Slices were perfused in a saline containing 200 microM sodium ascorbate. One-second puffs of ascorbate-free saline (zero [AA]o), delivered through a 2-3 microm diameter pipette, were directed at the bipolar cells. IK(V) was recorded by conventional whole-cell patch-clamp methods. In normal retinas, puffs of zero [AA]o caused a rapid (<100 ms) suppression of IK(V) of about 50% that lasted for several minutes. This effect was blocked by 1 microM SCH23390 and was unaffected by 2 mM Co2+ or 5 microM spiperone. 6-OHDA treatment resulted in major effects. First, IK(V) was reduced by approximately 50% for weeks 1-6, recovering to a 20% reduction by week 7. Second, puffs of zero [AA]o enhanced IK(V) rather than suppressed it. The enhancement was blocked by SCH23390 and the PKA inhibitor, Wiptide, but was insensitive to spiperone. Third, all parts of the Mb bipolar cell (except for the axon) were sensitive to puffs of zero [AA]o in both normal and 6-OHDA-treated retinas. Fourth, bath application of 20 microM dopamine restored the amplitude of IK(V) but did not reverse the effects of puffed zero [AA]o. IK(V) was fit by two exponentials; all of the effects on IK(V) were on the amplitude of the components and not on the time constants. Chronic dopamine depletion caused reversible changes in the properties of K+ channels underlying IK(V), as well as a long-term change in the intracellular coupling mechanisms between D1-receptor activation and the modulation of IK(V). PMID- 11417807 TI - The kinetics of hemopexin and alpha1-acid glycoprotein levels induced by injection of inflammatory agents in chickens. AB - The acute phase response to inflammation induces changes in the secretion of hepatic proteins. To examine the time course of an acute phase protein response in broiler chickens, the plasma levels of hemopexin (HX) and alpha1-acid glycoprotein (AGP) and liver HX mRNA were measured at various time points from 3 hr to 336 hr after an intraabdominal injection of either lipopolysaccharide (LPS), complete Freund's adjuvant, incomplete Freund's adjuvant, phytohemagglutin, or mineral oil. Uninjected chicks served as controls. The accumulation of liver HX mRNA began within 3 hr of stimulation and peaked at 12 hr. Relative to control levels, plasma HX and AGP levels increased by 6-12 hr postchallenge and peaked at 24 hr. Complete Freund's adjuvant and LPS treatments induced the greatest increase in plasma HX (threefold; P < 0.05). Plasma levels of HX and AGP returned to control levels at 336 and 168 hr postinjection, respectively. A second experiment demonstrated that turpentine induced a similar AGP response as LPS and that albumin is a negative acute phase protein. The results suggest that plasma levels of HX or AGP could be used as an indicator of the systemic component of a local inflammatory response in chickens. PMID- 11417808 TI - Molecular characterization of Brazilian infectious bursal disease viruses. AB - A reverse transcriptase-polymerase chain reaction (RT-PCR) procedure was used to amplify a VP2 gene fragment (248 bp) from infectious bursal disease virus (IBDV). The procedure allowed the detection of known IBDV strains from the United States, along with field isolates and commercial vaccines produced in Brazil. Amplified VP2 fragments were further characterized by restriction fragment length polymorphism (RFLP) analysis. From 55 Brazilian commercial flocks, 48 field samples were IBDV positive by RT-TCR. Vaccine RFLP patterns were found in 12 flocks, a pattern compatible with classic IBDV in one flock, four new patterns in 31 flocks, and a pattern compatible with very virulent (vv) IBDV in four flocks. Sequence analysis showed that the vvIBDV RFLP patterns were closely related to the vvIBDVs described in Europe and Asia. Phylogenetic analysis of the four new RFLP patterns showed that they were closely related to but distinct from other classic, variant, and vvIBDVs, suggesting a high prevalence of different IBDV strains in Brazilian commercial flocks. PMID- 11417809 TI - Blackhead disease (Histomonas meleagridis) aggravated in broiler chickens by concurrent infection with cecal coccidiosis (Eimeria tenella). AB - The effect of concurrent cecal coccidiosis infections on severity of Histomonas meleagridis (blackhead disease) in chickens was investigated in a series of experiments. Cecal lesions from H. meleagridis were severe in all inoculated control groups and did not appear to be affected by the introduction of Eimeria tenella infection. However, the severity of liver lesions and number of birds positive for liver lesions of H. meleagridis increased significantly with the presence of E. tenella. The increase was similar when 10(3) or 10(4) oocysts of E. tenella were given and was the same when oocysts were given at the same time as H. meleagridis or 4 days prior. The liver lesions increased directly as doses of H. meleagridis increased from 7.5 x 10(3) cells to 30, 100, or 300 x 10(3) when E. tenella was given along with H. melelagridis but not when H. meleagridis was given alone. Administration of a live coccidiosis vaccine containing very low levels of E. tenella also gave a significant boost to liver lesions but at a much lower level than that observed with larger doses of E. tenella. The positive relationship between infections of cecal coccidiosis and H. meleagridis in chickens suggests that such dual exposure may contribute to increased clinical outbreaks of blackhead disease in chickens under field conditions. PMID- 11417810 TI - The interaction between Newcastle disease virus and Escherichia coli endotoxin in chickens. AB - The interaction between Newcastle disease virus (NDV) and Escherichia coli endotoxin was studied in cell cultures, embryonated chicken eggs, and 8-wk-old chickens. These interactions were evaluated according to the induction of specific or nonspecific resistance in the host system and the virus titer produced in both chicken embryos and chickens. The endotoxin of E. coli induced a decrease in the size of the bursa of Fabricius in live chickens. Escherichia coli endotoxin given intravenously induced plasma antiviral activity in chickens that was interpreted to be interferon, as detected in a vesicular stomatitis virus plaque reduction assay. Endotoxin failed to produced toxic effects in the chicken embryo fibroblasts (CEFs) or to result in any antiviral effect because no change was noted in the number of NDV plaques formed in CEF cultures. When endotoxin was given 3 days before NDV exposure in chickens, the virus titers were significantly (P < 0.05) decreased from a peak of 10(2) to 10(0.18), 10(2.5) to 10(0.18), and 10(2.5) to 0 in the spleens, lungs, and kidneys, respectively, at 72 hr post-NDV inoculation. When endotoxin was given 24 hr after NDV inoculation, the NDV titer significantly (P < 0.05) increased from 10(2.0) to 10(3.5), 10(2.5) to 10(6.5), 10(2.5) to 10(4.5), 0 to 10(2.5) in the spleen, lungs, kidneys, and liver, respectively, at 72 hr after NDV inoculation. In chicken sera, hemagglutination inhibition (HI) titer to NDV was significantly (P < 0.05) enhanced from 1164 to 3127 when endotoxin was given prior to virus inoculation. However, there was a decrease in HI to NDV from 1164 to 727 without a significant difference in chicken sera when NDV was given prior to endotoxin inoculation. PMID- 11417811 TI - Mycoplasma gallisepticum in house finches (Carpodacus mexicanus) and other wild birds associated with poultry production facilities. AB - Since 1994, an epidemic of conjunctivitis caused by Mycoplasma gallisepticum (MG) has spread throughout the eastern population of house finches (Carpodacus mexicanus). The adaptation of MG to a free-flying avian species presents potential problems for the control of mycoplasmosis in commercial poultry. To evaluate risks associated with this emerging problem, a field survey was conducted to assess prevalence of MG infection in house finches and other passerine birds associated with poultry farms. Between November 1997 and March 1999, 1058 birds were captured by mist net or trap at 17 farms and at 10 feeder stations in northeast Georgia. Birds were bled and screened by serum plate agglutination (SPA) for antibodies to MG. Birds with negative or weak positive SPA results were released at capture sites, and those with strong positive SPA reactions were kept for further evaluation. Necropsies were performed on selected house finches and individuals of 11 other passerine species, and samples were collected for MG testing by culture, polymerase chain reaction (PCR), hemagglutination inhibition, and histopathology. Testing revealed 19.1% of 671 birds caught at farms and 11.6% of 387 birds caught at feeder sites were SPA positive for MG. Three house finches captured on farms were positive for MG by culture and PCR, whereas three from feeder sites were positive only by PCR. No MG isolates were made from tufted titmice (Baeolophus bicolor), but 40% were positive by PCR. Individuals from 10 additional species were SPA positive only. Results suggest that MG persists at low levels in house finches in northeast Georgia and that tufted titmice may be nonclinical carriers of MG or a related mycoplasma. Positive SPA reactions in other species may be caused by nonspecific reactions or contact exposure. Current biosecurity recommendations should be sufficient to minimize risks of transmission between wild and domestic birds. PMID- 11417812 TI - Amino acid comparison of infectious bursal disease viruses placed in the same or different molecular groups by RT/PCR-RFLP. AB - Infectious bursal disease virus (IBDV) strains have been identified and placed into molecular groups by a reverse transcriptase (RT)/polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) assay. The predicted amino acid sequences corresponding to the region of the genome examined by RFLP were determined and compared for 14 IBDV strains from different molecular groups and 11 IBDV strains that were identified in molecular group 6. Among the viruses within molecular group 6, 13 amino acid positions had mutations, and among the viruses in different molecular groups, 27 amino acid positions had mutations. In addition to having more mutations, viruses compared from different molecular groups also had mutations at key positions that were previously reported to be important for the formation of neutralizing epitopes. Three of these IBDV strains with unique RFLP patterns were used to challenge 1-wk-old broiler chickens with maternal immunity to IBDV. One of these viruses, T1, broke through this maternal immunity as evidenced by detection of the virus by RT/PCR-RFLP and production of an active virus neutralizing antibody response to classic and variant IBDV strains. Unique amino acid mutations in the T1 virus that may have contributed to its ability to break through this maternal immunity were observed at amino acids 318 and 322. The results indicate that RFLP profiles and nucleotide sequences can be used to predict the relative similarities and differences among IBDV strains, but determining the actual antigenic differences among viruses requires testing in vivo. PMID- 11417813 TI - Study of protection by recombinant fowl poxvirus expressing C-terminal nucleocapsid protein of infectious bronchitis virus against challenge. AB - A stable recombinant fowl poxvirus (rFPV) expressing the C-terminal region (119 amino acids) of the nucleocapsid (N) protein of an infectious bronchitis virus (IBV) strain Ch3 was constructed by inserting the coding sequence within the thymidine kinase gene of fowl poxvirus (FPV) by homologous recombination. The N protein was expressed under control of the vaccinia virus promoter P7.5 in chicken embryo fibroblast cell cultures as seen in immunofluorescence assay and in rFPV-inoculated specific-pathogen-free (SPF) chickens by detecting antibodies with enzyme-linked immunosorbent assay (ELISA). A homologous IBV strain (Ch3) and two heterologous IBV strains (Ch5 and H4) were used to inoculate SPF chickens in a challenge to examine the protective efficacy of the rFPV. When the chickens were challenged with IBV Ch3 or Ch5, the control birds had respiratory signs of infections bronchitis, whereas all the vaccinated birds were clinically normal although low levels of the IBV infection were detected by a differential ELISA. In contrast, in the chickens challenged with IBV H4, all control birds and vaccinated birds suffered from the highly lethal IBV H4 infection. Our results suggest that the C-terminal 119 amino acid of the nucleocapsid expressed by FPV is a host-protective antigen and may induce cross-protective immunity against illness among some IBV strains. PMID- 11417814 TI - The influence of crumble and mash feed on oral lesions of white leghorn laying hens. AB - The effect of crumble and mash feed on oral lesions in laying hens was investigated. Each of two groups of white leghorn hens (group A, n = 120; group B, n = 120) was separated into crumble- and mash-fed treatments. At the start of the trial, group A chickens were free of oral lesions, whereas group B chickens had nearly 100% prevalence of oral lesions. The trials began at 17.4 and 22 wk of age and ended at 32 and 33 wk of age for groups A and B, respectively. Chickens were housed in cages indoors, fed free choice, and serologically monitored. Oral lesions were periodically evaluated by visual scoring. Lesion prevalence peaked in group A chickens at 23-25 wk of age, concurrently with an outbreak of respiratory disease, and decreased to a low level at the end of the trial. For most of the trial, crumble-fed chickens had significantly lower oral lesion prevalence than mash-fed chickens (P < 0.05). Lesion prevalence in group B chickens decreased after treatments began and maintained this trend to the end of the trial. Compared with mash-fed chickens, crumble-fed chickens in group B had significantly lower oral lesion prevalence throughout the trial (P < 0.001). Neither group had significant differences in oral lesion scores between treatments. Most lesions were on the lower palate, pinpoint to 2 mm in diameter, often multiple, and bilaterally symmetrical. Feed analyses for mycotoxins and heavy metals were within normal limits. The inciting etiology of oral lesions was not elucidated in this trial. However, this work demonstrated that once lesions were present, feed type had a strong effect on persistence of oral lesions. PMID- 11417815 TI - Efficacy of vaccines in chickens against highly pathogenic Hong Kong H5N1 avian influenza. AB - In 1997, highly pathogenic (HP) H5N1 avian influenza virus (AIV) caused infections in poultry in Hong Kong and crossed into humans, resulting in a limited number of infections including 18 hospitalized cases and six associated deaths. The unique ability of this, AIV to infect both poultry and people raised a concern for the potential of humans to be biological as well as mechanical vectors of this AIV to poultry. The current study was undertaken to determine if existing vaccines and their technologies could be used during an outbreak to protect poultry. Commercial and experimental inactivated whole H5 AIV and baculovirus-expressed AIV H5 hemagglurinin protein vaccines provided protection from clinical signs and death in chickens after lethal challenge by human-origin HP H5N1 Hong Kong strains 156/97 and 483/97. The commercial and experimental inactivated vaccines had mean protective doses ranging from 0.25 to 0.89, which represents the milligrams of viral protein in the vaccines that provided protection from death in half of the birds. Furthermore, the vaccines reduced the ability of the challenge AIV to replicate in chickens and decreased the recovery of challenge AIV from the enteric and respiratory tracts, but the use of a vaccine will nor totally prevent AI virus replication and shedding. Existing vaccines will protect poultry from mortality and reduce virus replication from the new HP AIV strain that can infect both poultry and humans. PMID- 11417816 TI - Spike glycoprotein cleavage recognition site analysis of infectious bronchitis virus. AB - The spike glycoprotein of infectious bronchitis virus (IBV), a coronavirus, is translated as a precursor protein (So), then cleaved into two subunits (S1 and S2) by host cell serine proteases. In this study, we compared the cleavage recognition site of 55 IBV isolates to determine if the cleavage recognition site sequence, which consists of five basic amino acid residues, correlates with host cell range, serotype, geographic origin, and pathogenicity as it does in orthomyxoviruses and paramyxoviruses. The most common cleavage recognition site observed (33 of 55 viruses) was Arg-Arg-Ser-Arg-Arg, representing at least 11 different serotypes. Thus, cleavage recognition site does not appear to correlate with serotype. We also determined that cleavage recognition site sequence does not correlate with pathogenicity because attenuated and pathogenic isolates (different passages of the same virus) contain identical cleavage recognition site sequences. In addition, nephropathogenic strains had the same cleavage recognition site sequence as many nonnephropathogenic isolates. Cleavage recognition site sequence does correlate with viruses in different geographic regions, which may be an important characteristic to examine in epidemiologic studies. An IBV monoclonal antibody neutralization-resistant mutant (NR 18) had an unusual substitution of Ile for Arg at the fourth position, giving the sequence Arg-Arg-Ser-Ile-Arg, which likely prevents cleavage and, thus, destroys the conformationally dependent monoclonal antibody binding epitope. Six residues on the amino-terminal side of the cleavage recognition site are conserved in 31% of the isolates and consist of only one or two basic amino acids. Thus, the number of basic residues around the cleavage recognition site does not appear to correlate with increased cleavability, host cell range, and increased virulence as it does with envelope glycoproteins in orthomyxoviruses and paramyxoviruses. PMID- 11417817 TI - Genetic characterization of chicken anemia virus from commercial broiler chickens in Alabama. AB - Chicken anemia virus (CAV) isolates show extremely limited genetic variability worldwide. We determined the nucleotide sequence of an 823-nucleotide portion of the 2.3-kb CAV genome found in 10 liver and/or spleen specimens of Alabama 29-to 49-day-old commercial broiler chickens exhibiting lymphocyte depletion of the thymus submitted to the state diagnostic laboratory because of problems unrelated to anemia. We determined the nucleotide sequence directly from DNA isolated from tissues, without isolation of virus in culture. This procedure enabled us to characterize CAV that might not have replicated in culture and avoided the potential for changes during passage. Results confirmed the limited genetic variability of CAV. All sequences were identical in 93% of nucleotide positions. The sequences encoded only two distinct VP1 hypervariable regions, and both had been found previously in other CAV isolates. A novel amino acid, glutamine, was found at VP1 position 22 in half the sequences, replacing the histidine residue encoded by most previously characterized CAV genomes. We were able to distinguish among CAV genomes with different codons at VP1 amino acid 22 and different hypervariable regions by restriction endonuclease analysis of polymerase chain reaction products. PMID- 11417818 TI - The in vivo and in vitro effects of chicken interferon alpha on infectious bursal disease virus and Newcastle disease virus infection. AB - The in vitro and in vivo effects of chicken interferon alpha on infectious bursal disease virus (IBDV) infection were investigated in this study. A cDNA of interferon alpha was first cloned from a Chinese strain chicken Shiqi by reverse transcription-polymerase chain reaction. The deduced amino acid sequence has one amino acid substitution with chicken interferon alpha 1 at residue 65 (N to S) and two amino acid substitutions with chicken interferon alpha 2 at residues 50 (N to S) and 58 (P to L), respectively. A prokaryotic expression system was employed to produce a large quantity of recombinant protein. Recombinant interferon was purified in a one-step process, and an optimal refolding process was devised. About 51% recombinant protein from inclusion bodies was refolded, and the final yield of the recombinant interferon reached 24.66 mg/liter culture. The recombinant interferon suppressed IBDV plaque formation in a dose-dependent manner and ameliorated IBDV and Newcastle disease virus infection in both specific-pathogen-free (SPF) and commercial chickens. The antiviral effect of interferon alpha is more significant in commercial chickens than in SPF chickens, and the route of administration affects the efficacy of interferon therapy. This is the first reported study of the effects of interferon alpha on IBDV infection. PMID- 11417819 TI - Genetic variation in susceptibility to Marek's disease in a commercial broiler population. AB - Two commercial broiler pure lines that were previously identified to differ in their susceptibility to Marek's disease (MD) were line-crossed to generate an F1 population. Eight F1 males were randomly mated to four or five F1 females to produce an F2 test population that would be segregating for genes affecting MD. All F2 progeny (four hatches) were pedigreed at hatch and placed in colony houses as nonvaccinated. At 5 days of age, they were challenged intraabdominally with MD virus RB1B. Clinical signs, mortality, and gross and microscopic lesions were recorded during the MD challenge. At 8 wk postchallenge, all remaining birds were euthanatized and necropsied. During the MD challenge of the first two hatches, we observed that several severely stunted broilers originated from certain families and the differences in body weight among birds appeared as early as 3 wk postchallenge. To confirm this observation, body weight at 6 wk postchallenge was determined for all surviving birds in hatches 3 and 4 (n = 242). Genetic variation in body weight among broiler sire families was apparent; the average body weight for males at this time was 2.07 kg, whereas with females, it was 1.87 kg. At least 12.2% of the broilers, including both sexes, weighed less than 1 kg ("severely stunted") at this time. The incidence of these growth-stunted birds within each broiler sire family ranged from 0 to 26% and for dam families, 0 to 60%. Correlation analyses between stunting and other MD-associated traits revealed that the incidence of stunting had a significant and positive association with paralysis (r = 0.50). Therefore, the data suggest that there may be a genetic component affecting body weight loss during MD infection. The genetic component is speculated to affect susceptibility to MD paralysis with an indirect effect on the body weight of birds. The significance of this finding is best exemplified by the identification of a broiler sire family with over 26% of its progeny affected by this MD-associated trait. PMID- 11417820 TI - Detection and screening of Salmonella enteritidis-infected chickens with recombinant flagellin. AB - Screening and identification of Salmonella enteritidis in commercial poultry flocks have assumed principal roles in preventing transmission of this pathogen to humans from hen eggs. Serologic diagnosis of S. enteritidis infection in commercial flocks currently relies on laboratory-based tests for detection of antibodies to the lipopolysaccharide, whole flagella, and bacteria. We amplified a sequence from the g,m flagellin of S. enteritidis, followed by cloning, expression, and purification of the protein. The recombinant protein was first characterized by western blot and subsequently evaluated as enzyme-linked immunosorbent assay (ELISA) antigen for detection of S. enteritidis infection. A total number of 49 positive sera and 40 negative sera were tested for ELISA validation. A cutoff value of 0.14 was shown to be sufficient to discriminate the negative and positive sera. Results obtained by testing sera raised against different bacterial strains/serotypes further confirmed that this recombinant flagellin-based ELISA was indeed specific for the detection of S. enteritidis. Both sensitivity and specificity of the developed ELISA test were comparable with a commercially available test, indicating that it is a highly promising and reliable diagnostic tool for S. enteritidis infection. PMID- 11417821 TI - Characterization of three infectious bronchitis virus isolates from China associated with proventriculus in vaccinated chickens. AB - Outbreaks of an avian disease in infectious bronchitis-vaccinated chickens in China have led to the characterization of coronaviral isolates Q1, J2, and T3, which were isolated from proventricular tissues of the affected young layer flocks. Serologic analysis revealed that they could induce high titers of infectious bronchitis virus (IBV) antibodies in inoculated specific-pathogen-free (SPF) chickens in indirect enzyme-linked immunosorbent assay but were not neutralized by antisera specific to the IBV serotype M41 and the Australian T strain. In a pathogenicity experiment, the clinical signs and related gross lesions resembling those of field outbreaks were reproduced in SPF chickens, and viruses were reisolated from the damaged tissues, including trachea, proventriculus, duodenum, and cecal tonsil. Sequence data demonstrated the complete S1 amino acid sequences of these isolates were almost identical despite recovery from geographically different areas in China and had 47.3%-82.3% similarity in comparison with the 47 published S1 sequences. On the basis of genotyping and limited serology, the three isolates, which were responsible for field outbreaks of the disease, might be a new IBV variant. PMID- 11417822 TI - The relationship between the magnitude of the specific antibody response to experimental salmonella enteritidis infection in laying hens and their production of contaminated eggs. AB - Detecting infected laying flocks is a vital part of many efforts to control egg associated transmission of Salmonella enteritidis to humans. The relationship between the development of a specific antibody response in infected hens and the deposition of S. enteritidis in eggs is important for establishing the epidemiologic relevance of serologic testing methods. In two trials, laying hens were infected with large oral doses of phage types 13a and 14b isolates of S. enteritidis. Approximately 38% of all infected hens produced at least one contaminated egg, at an overall incidence of 5.2%, between 3 and 23 days postinoculation. As determined by enzyme-linked immunosorbent assay with an S. enteritidis flagellar antigen, 91.7% of inoculated hens produced specific serum antibodies. Although hens with very high antibody titers were associated with a significantly elevated frequency of egg contamination, a consistently direct relationship was not evident between the magnitude of the antibody responses of individual hens and the frequency at which they laid contaminated eggs. Accordingly, although serologic tests can be valuable screening tools for preliminary detection of S. enteritidis infections in poultry, the magnitude of the antibody responses detected in individual hens may not predict the overall risk of egg contamination associated with particular laying flocks. PMID- 11417823 TI - Comparison of heterophil phagocytosis for heterophil-adapted Salmonella enteritidis (HASE) and wild-type Salmonella enteritidis (SE). AB - Serial passage of Salmonella enteritidis (SE) yields heterophil-adapted SE (HASE) strains that have resulted in decreased shedding of SE in feces and reduced egg contamination. Additionally, increasing the number of heterophil passages further reduced the number and frequency of fecal shedding. To evaluate SE and heterophil interaction, nine SE strains were fluorescein isothiocyanate-labeled when viable. There were six wild-types: SE TK 474, SE TK 584, SE TK 599, SE TK 600, SE TK 655, and SE TK 657; and three HASE strains: TK 499 heterophil adapted five times, TK 598 heterophil adapted six times, and TK 605 heterophil adapted 11 times. Trials were repeated seven times in duplicate with heterophils isolated from seven healthy chickens. Heterophils were incubated with the bacterial strains at 41 C for 15 min, and 10,000 heterophils were analyzed by flow cytometry. Percentage of phagocytosis and mean channel number of fluorescence were compared. Both parameters were significantly increased for all HASE-type strains compared with wild-type, nonadapted SE strains. Increased phagocytosis of HASE bacterial strains may be significant in processing and elimination of the HASE strains and may be related to the protective effect of HASE by decreased shedding of wild type SE challenge strains. PMID- 11417824 TI - A novel delivery of oxytetracycline in turkey breeder hens. AB - A novel product (SQ12) for subcutaneous (SQ) injectable delivery of oxytetracycline (OTC) has been developed for use in livestock. SQ12 employs microfluidic spheres encasing OTC crystals, which allows for longer release of the OTC compared with other injectable antibiotics. The objectives of the study were to determine serum and tissue levels of SQ12 in turkey breeder hens to 14 days postinjection and to evaluate effects of SQ12 on reproductive status. Thirty photostimulated hens were housed in litter floor pens and provided with 14.5 hr of light per day in a curtain-sided facility. Six hens served as untreated controls. Twelve hens per treatment group received SQ injections in the neck with SQ12 at 11.4 (L dose group) or 22.7 mg/kg (H dose group) to assess low and high doses, respectively. Serum samples were obtained from each hen at predose and 6, 12, 24, 48, 72, 96, 168, 240, and 336 hr postinjection. All hens were euthanatized at 14 and 15 days postinjection. One-half of the hens in each treatment group were sampled (liver, lung, kidneys, and breast muscle) for tissue residue levels of OTC. The control group had no detectable OTC in serum or tissues at any sample collection time. There were no detectable serum levels of OTC in either treatment group prior to injection. The average serum concentrations of the L and H dose groups showed similar depletion curves although the H dose group was 42% higher at maximum concentration than the L group. Average tissue concentration of OTC for all tissues sampled from the H dose group was twice that of the L dose group. All tissue levels were below the OTC residue tolerance limit. SQ12 provided an extended source of OTC in serum of turkey breeder hens with no effect on reproductive status. SQ12 may provide for a novel treatment of bacterial infection in turkey breeder hens with longer lasting serum levels compared with other single injectable OTC products. PMID- 11417825 TI - Adenovirus pathogenicity in immature ostriches. AB - Two group I avian adenoviruses implicated as the possible cause of "fading chick syndrome" in ostriches less than 8 wk of age were isolated in primary chicken embryo liver cells. These viruses were identified by virus neutralization and further characterized by a pathogenicity trial in immature ostriches. The results showed that these isolates were noninfectious in ostrich chicks. PMID- 11417826 TI - Bacterial flora of the conjunctiva and nasal cavity in normal and diseased captive bustards. AB - A survey was carried out to describe the normal aerobic bacterial flora of the conjunctiva and nasal cavity of captive houbara bustards (Chlamydotis undulata), kori bustards (Ardeotis kori), and white-bellied bustards (Eupodotis senegalensis) maintained at the National Avian Research Center, Abu Dhabi, United Arab Emirates. A total of 58 samples were examined from the nasal cavity and 55 samples from the conjunctiva of healthy bustards. There was no bacterial growth in 45% of conjunctival samples. Bacteria isolated from the conjunctiva of healthy birds included Micrococcus spp., Staphylococcus auricularis, Staphylococcus xylosus, Staphylococcus capitis, Staphylococcus warneri, Bacillus spp., and Enterobacter amigenus. Bacteria isolated from the nasal cavity of healthy birds included Bacillus spp., Micrococcus spp., S. auricularis, S. xylosus, Staphylococcus simulans, Staphylococcus saprophyticus, Staphylococcus hyicus, Staphylococcus cohnii, Staphylococcus sciuri, Aerococcus spp., and Providencia rettgeri. These findings were compared with bacterial isolates from bustards with clinical signs of ocular or upper respiratory tract diseases. Mycoplasma spp., Pseudomonas aeruginosa, Pseudomonas stutzeri, Proteus mirabilis, Escherichia coli, Klebsiella spp., Aeromonas hydrophila, and Staphylococcus aureus were the pathogenic bacteria isolated from the conjunctiva of 34.3% bustards with ocular discharges. Mycoplasma spp., P. aeruginosa, Pseudomonas spp., P. mirabilis, E. coli, Klebsiella pneumoniae, and S. aureus were the pathogenic bacteria isolated from the nasal cavity of 74% bustards with upper respiratory tract diseases. PMID- 11417827 TI - Loss of virulence by heterophil-adapted Salmonella pullorum. AB - We report that reduction of virulence for day-old chicks was achieved after eight times-repeated heterophil passage of Salmonella pullorum (SP). The virulent source strain SP-V caused 64% mortality and 89% internal organ as well as 89% cecal colonization 10 days after administration of 10(7) colony-forming units (CFU) to day-old chicks. Eight-times-repeated passage of SP in heterophils resulted in attenuated strain SP-A that was nonlethal and reduced colonization of internal organs from 89% for SP-V strain to 4.3% for SP-A strain 10 days after administration of 10(7) or 10(8) CFU to day-old chicks. Cecal colonization was reduced from 89% for SP-V strain to 0 for SP-A strain 10 days after administration of 10(7) or 10(8) CFU to day-old chicks. PMID- 11417828 TI - In vitro susceptibility of avian mycoplasmas to enrofloxacin, sarafloxacin, tylosin, and oxytetracycline. AB - In vitro susceptibility of avian Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) to enrofloxacin, sarafloxacin, tylosin, and oxytetracycline was determined by a serial broth dilution method. The minimum inhibitory concentration (MIC) was recognized by a conversion of the pH indicator phenol red in culture media to a yellow color. Each isolate or type strain of mycoplasma was tested in two replicates. The MICs of tylosin, enrofloxacin, sarafloxacin, and oxytetracycline against five isolates and two reference strains of MG (approximately 10(5) colony-forming units [CFU]/ml) were 0.05, 0.14, 0.37, and 1.30 microg/ml, respectively. The MICs of the four antimicrobial agents against six isolates and one reference strain of MS (approximate 10(5) CFU/ml) were 0.13, 1.82, 1.76, and 0.91 microg/ml, respectively. There were no differences (P > 0.05) between tylosin, enrofloxacin, and sarafloxacin against MG, but these three antibiotics were different (P < 0.05) from oxytetracycline. The MIC value of tylosin against MS was different (P < 0.05) from those of sarafloxacin and enrofloxacin, but it was not different (P > 0.05) from that of oxytetracycline. PMID- 11417829 TI - Relationship of increased fowl cholera outbreaks in turkeys with high environmental temperatures. AB - The relationship of an increase in fowl cholera outbreaks in turkeys with an increase in environmental temperatures during June, July, August, and September between 1959 and 1992 was analyzed. High environmental temperatures were found to be influential in the development of fowl cholera in turkeys. When the average monthly maximum environmental temperatures for 5 mo of July and 7 mo of August during the 13 yr between 1967 and 1979 were above 30.5 C, there was a significantly (P < 0.05) higher number of fowl cholera outbreaks in turkeys for each month than during the same months when the average maximum temperatures were below 30.5 C. To test the hypothesis that an increase in fowl cholera outbreaks was preceded by an increase in temperature, the pre- and postoutbreak temperatures for 46 selected outbreak clusters occurring between 1959 and 1992 were averaged. Both the average maximum and minimum temperatures for the latter 9 days of the preoutbreak period were highly significantly (P < 0.001) higher than those of the average cluster outbreak day and the following four postoutbreak days. Also, for the nine individual days of the latter pre-outbreak period, the daily average maximum temperature was significantly (P < 0.05) higher for 3 days and partially significantly (P < 0.10) higher for 3 days than that of the average cluster outbreak day, and the daily average minimum temperature was significantly (P < 0.05) higher for 2 days and partially significantly (P < 0.10) higher for 1 day than that for the average cluster outbreak day. PMID- 11417830 TI - Plasma biochemistry in pigeons experimentally infected with Salmonella. AB - Ten pigeons were crop inoculated with 1 x 10(9) colony-forming units of Salmonella typhimurium var. Copenhagen and observed during 28 days. Ten sham inoculated pigeons served as noninfected controls. Clinical signs after Salmonella infection consisted of polydipsia, polyuria, and diarrhea. Morbidity was 90%, but there was no mortality. All inoculated pigeons showed fecal excretion of Salmonella for at least 7 days. Biochemical analysis of plasma samples taken at 3-day intervals indicated decreased concentrations of creatine kinase (CK)-MM and CK-MB isoenzymes and elevated total protein and alpha- and gamma-globulin values. No consistent changes in the level of 17 other blood parameters were observed. After 28 days, all pigeons were necropsied. Gross lesions and bacteriologic and histologic examination indicated septicemia in all Salmonella-inoculated pigeons. Results indicate that Salmonella septicemia in pigeons induces only limited changes in biochemical blood parameters. Decreased CK concentration was a consistent finding, however, and may therefore be a useful aid in the diagnosis of salmonellosis in pigeons. PMID- 11417831 TI - In vivo biologic effects of recombinant-turkey interferon-gamma in neonatal leghorn chicks: protection against Salmonella enteritidis organ invasion. AB - Interferon-gamma (IFNgamma) has been demonstrated to have potent stimulatory effects on parameters of cell-mediated immunity in chickens (11). Protection of neonatal leghorn chickens against infection by invasive salmonellae has been associated with enhanced cell-mediated indices of immunity (5). The present investigation evaluated the effect of recombinant-turkey (rt) IFN-gamma on protection of neonatal leghorn chicks from Salmonella enteritidis (SE) organ invasion after experimental challenge in three experiments. In Expt. 1, intraperitoneal (i.p.) administration of 25 microg rtIFNgamma per chick 30 min prior to per os SE challenge resulted in a 35% reduction (P < 0.01) in SE organ invasion when compared with control (vehicle injected) chicks 24 hr post-SE challenge. However, i.p. administration of 2.5 microg rtIFNy per chick was not efficacious in reducing SE organ invasion. In Expt. 2 and Expt. 3, i.p. administration of 13.75 microg rtIFNgamma per chick 30 min prior to per os SE challenge resulted in significant reductions of 38.4% (P < 0.025) and 31.58% (P < 0.01), respectively, in SE organ invasion as compared with control chicks 24 hr post-SE challenge. Administration of 2.5 or 25 microg rtIFNgamma per chick i.p. had no effect on SE organ invasion in either Expt. 2 or Expt. 3 24 hr post-SE challenge. Additionally, i.p. administration of rtIFNgamma 30 min prior to SE challenge in Expt. 2 and Expt. 3 was not associated with protection against SE organ invasion when organ culture was performed 72 hr postchallenge. Further, the oral administration of 25 microg rtIFNgamma per chick was not efficacious in conferring protection against SE organ invasion at 24 or 72 hr postchallenge when this route of administration was evaluated in Expt. 2. Similarly, the subcutaneous administration of a potential repository injection of 13.75 or 25 microg rtIFNgamma per chick did not protect chicks against SE organ invasion when evaluated 72 hr postchallenge. These data indicate a potential acute immunostimulatory activity of rtIFNgamma in chickens experimentally challenged with SE. Further, these experiments, although preliminary, are suggestive of the potential involvement of IFNgamma in cell-mediated or innate mechanisms of protective immunity against salmonellosis in chickens. PMID- 11417832 TI - Effect of prolonged heat stress in single-comb white leghorn hens on progeny resistance to Salmonella enteritidis organ invasion. AB - In our laboratory, we have often had difficulty infecting neonatal chickens with invasive salmonellae when ambient temperatures exceed 30 C. We hypothesized that this increased resistance in chicks during warmer months may be associated with heat stress-associated maternal factors. Presently, single-comb white leghorn hens were separated into a non-heat-stressed group, reared under temperatures from approximately 10 to 24 C, and a heat-stressed group, in which environmental temperature was incrementally elevated to near 37 C and maintained for the duration of the 13-wk study. For Expt. 1, eggs from heat-stressed or control hens, collected on days 8-14 of the study, were pooled respective to treatment and incubated. At the time of egg collection, mean hen-day egg production was 51.83% or 65% for heat-stressed or control hens, respectively. On day of hatch, progeny from hens in each group were orally challenged with 0.9 x 10(4) colony forming units (CFU) Salmonella enteritidis (SE). Rates of SE organ invasion of 97.3% or 94.4% were obtained in progeny from heat-stressed or control hens, respectively. In Expt. 2, eggs from heat-stressed or control hens from days 30-42 of the study were collected and pooled by treatment for incubation. Mean hen-day egg production was 46.5% or 72.85% for heat-stressed or control hens, respectively. On day of hatch, progeny were orally challenged with either 2.2 x 10(3) or 2.2 x 10(4) CFU SE. A 100% incidence in SE organ invasion was observed in all groups. In Expt. 3, eggs were collected from days 43 through 56 of the study. Mean hen-day egg production was 19.8% or 76.8% for heat-stressed or control hens, respectively. On day of hatch, progeny were orally challenged with 2 x 10(3) CFU SE. Rates of SE organ invasion of 95.8% or 95.6% were obtained in progeny from heat-stressed or control hens, respectively. These data suggest that factors other than elevated temperature may be responsible for seasonal resistance to invasive salmonellae infection in neonatal chickens observed in our laboratory during warmer months in Texas. PMID- 11417833 TI - Serologic reactions against Salmonella in samples from broiler parent stock with and without preceding colibacillosis: a case-control study. AB - In the Danish Salmonella Control Program, eggs from broiler parent flocks are surveyed by serologic analysis every 4 wk for antibodies against Salmonella lipopolysaccharide O-antigens 1, 4, 5, 9, and 12 (Mix-enzyme-linked immunosorbent assay [ELISA]) and 6 and 7 (Infantis-ELISA). The antibody response is measured in percentage optical density (OD%) of a strong positive reaction, and the cutoff value has been determined to be 40 OD%. Two or more reactors above 40 OD% will place the parent flock under suspicion. There has been concern about possible cross-reactions between Salmonella spp. and other Enterobacteriaceae, e.g., Escherichia coli, because a high specificity of a Salmonella antibody test is desirable. Moreover, false-positive Salmonella results have economic consequences and impede planning the production. A case-control study based on cases of clinical E. coli infections (colibacillosis) from two Danish hatcheries, supplying about 62% of the Danish broiler production, is described. In order to eliminate a possible bias from age and season, the controls were matched on age of the birds and on time of submitting the samples. This study shows that flocks with preceding colibacillosis did not have higher salmonella reactions than matched flocks without a preceding colibacillosis. This observation was confirmed in longitudinal studies. PMID- 11417834 TI - Molecular characterization of infectious bronchitis virus isolates foreign to the United States and comparison with United States isolates. AB - Eleven infectious bronchitis virus (IBV) isolates foreign to the United States were analyzed by using reverse transcriptase (RT)-polymerase chain reaction (PCR)/restriction fragment length polymorphism (RFLP) and S1 glycoprotein gene sequencing. Two of the isolates generated RFLP patterns that resembled the Mass 41 strain. Seven novel RFLP patterns were detected among the other nine foreign IBV isolates. Five of the foreign isolates were further analyzed by S1 glycoprotein gene sequencing in our laboratory. Phylogenetic analysis of S1 glycoprotein-deduced amino acid sequences for 4/91 pathogenic, 4/91 attenuated, and Variant 1 were greater than 90% similar to viruses belonging to the 793/B serogroup and, therefore, are possibly serologically related. Variant 2 was only 81.0% similar to viruses belonging to the European serogroup B, and, therefore, predicting its serotype is difficult. Isolates 98-07484 and 97-8123 were genotypically unique and therefore might be serologically unique. With the RFLP patterns and the deduced S1 amino acid sequence data as a reference, none of the IBV isolates foreign to the United States have been detected in the United States. PMID- 11417835 TI - Presence and recovery of Ascaridia dissimilis ova on the external shell surface of turkey eggs. AB - Ascaridia dissimilis, a roundworm in turkeys, has been noted with increased frequency in commercial turkeys. Because infected turkeys can shed A. dissimilis ova in their feces, the potential exists for the external surface of turkey eggshells to be contaminated with A. dissimilis ova. The objectives of this study were to determine the presence of and recover A. dissimilis ova on the external surface of the turkey egg. In Experiment 1, turkey eggs were collected from naturally infected flocks, and eggs were processed by a sodium hydroxide procedure to recover any A. dissimilis ova on the external egg surface. In Experiment 2, the external surface of the turkey eggs was inoculated with 116 A. dissimilis ova/g feces, and eggshells were sampled every 3 days until 28 days of incubation to assess the recovery of A. dissimilis ova from the eggshell. In Experiment 1, of the 36 eggs examined from a flock naturally infected with A. dissimilis, one egg had an A. dissimilis ovum on its external eggshell surface. Experiment 2 demonstrated that A. dissimilis ova can be recovered from the external egg surface after a 28-day incubation period in the incubator. Ova recovery declined from an average of 62 A. dissimilis ova/turkey egg at day 2 of incubation to an average of 3 A. dissimilis ova/turkey egg at day 28 of incubation. PMID- 11417836 TI - In situ localization of infectious bursal disease virus-binding cells by a biotin streptavidin system. AB - A biotin-streptavidin system was established to directly visualize infectious bursal disease virus (IBDV)-binding cells in cell culture or in fresh tissues. The cells or tissue sections were first incubated with a biotinylated, purified IBDV strain GZ911 and then with a streptavidin-beta-galactosidase conjugate. In the presence of the enzyme substrate X-gal, IBDV-binding cells were labeled in blue color. By applying this method to frozen tissue sections, virus-binding sites were localized in situ in the bursa, spleen, and kidney tissue sections, whereas no positive cells were detected in the thymus tissue sections. Chicken embryo fibroblasts, Vero cells, MOP-8 cells, 293-EBNA cells, PANC-1 cells, and HuTu 80 cells were found to bind to the virus. However, the binding of the virus to MDA-MB-231 cells and SVG p12 cells was undetectable. This method can be employed for the expressional cloning of IBDV receptor and can be applied to studies on other avian viruses. PMID- 11417837 TI - Selection of thermostable Newcastle disease virus progeny from reference and vaccine strains. AB - In a study of low-virulence Newcastle disease virus (NDV) isolates from poultry, 38% of the isolates had a more thermostable hemagglutinin than the lentogenic reference strains B1 and La Sota or live vaccines derived from those strains. Whether those strains with a more thermostable hemagglutinin are truly indigenous or whether they could have originated from vaccines used in the flocks was unknown. Seven monovalent NDV vaccines of B1 or La Sota type and reference B1 and La Sota strains were heat treated at 56 C to select variants more thermostable than the parent virus. Four thermal treatment cycles were completed, and virus propagated from the second and fourth heat treatments was assayed for changes in thermostability and antigenicity. The hemagglutinin thermostability of all vaccine and reference strain variants increased from the initial < or =10 min to > or =120 min after four treatments. Antigenic changes evaluated by hemagglutination inhibition against NDV monoclonal antibodies identified changes in only the heat-treated La Sota strains. The results demonstrate that the field isolates with a more thermostable hemagglutinin could have been derived by selection from the heterogenous NDV populations in vaccine strains and that minor antigenic changes may be a result of that selection. PMID- 11417838 TI - No effect of turkey herpesvirus infections on the outcome of avian pneumovirus infections in turkeys. AB - To determine whether turkey herpesvirus (HVT) impairs the aspecific and specific defense against an avian pneumovirus (APV) infection, specific-pathogen-free turkeys were inoculated at 7 days of age with HVT and 1, 5, or 7 wk later with APV. Clinical signs, APV replication, and development of antibodies against APV were evaluated. No differences were found between the birds that received both HVT and APV and those that received only APV. PMID- 11417839 TI - An outbreak of duck viral enteritis (duck plague) in domestic Muscovy ducks (Cairina moschata domesticus) in Illinois. AB - Duck viral enteritis (DVE) was diagnosed in an outbreak of the disease in a resident population of Muscovy ducks (Cairina moschata domesticus) on a privately owned multispecies game bird production facility in Illinois, where it claimed 625 ducks. This disease condition had not been reported previously in domestic ducks in Illinois. Although other varieties and age groups of domestic waterfowl (i.e., black ducks, rhumen ducks, Pekin ducks, ducklings, and geese) were present on the game bird farm, the morbidity and mortality (100%) in this epornitic was solely limited to adult ducks of the Muscovy lineage. The clinical signs in the affected ducks were lethargy, diarrhea, dehydration, and death within 2-3 hr of onset of symptoms. Gross pathologic changes were nonspecific and included ecchymotic hemorrhage, effusion of fluid and blood within body cavities reflective of an acute systemic infectious disease. Light microscopic findings were necrosis of primarily digestive lining epithelium and variable lymphohistiocytic infiltration within mucosal and serosal connective tissues. Intranuclear inclusions resembling characteristic herpetic (i.e., Cowdry type A) inclusions were observed primarily in the digestive, respiratory, and reproductive tracts; liver; and spleen. Esophageal candidiasis, bacteriosis, and systemic Pasteurella anatipestifer infections, thought to be concurrent or opportunistic infections, were present in several ducks. DVE virus was demonstrated in infected Muscovy duck embryo fibroblast cells by direct DVE virus specific fluorescent antibody staining. PMID- 11417840 TI - Detection of chicken anemia virus DNA in the thymus of naturally infected chicks in turkey. AB - In this study, 94 clinically ill 11-28-day-old chicks belonging to eight broiler units from the Marmara region were investigated clinically for changes in hematocrit values and for the presence of chicken anemia virus (CAV) DNA. CAV DNA was detected by polymerase chain reaction in the thymus of 8.5% of the chicks. These chicks showed clinical signs of diarrhea, anorexia, depression, and growth retardation. The hematocrit values of these chicks were between 24% and 38%. At necropsy, hemorrhages were observed in the leg and pectoral muscles. Atrophy was noted in the thymus and in the bursa of Fabricius of positive chicks, and hemorrhages in the proventriculus of one positive chick were observed. This report describes the first detection of CAV DNA in chicks in Turkey. PMID- 11417841 TI - Monitoring Mycoplasma gallisepticum and Mycoplasma synoviae infection in breeder chickens after treatment with enrofloxacin. AB - Three experimental strains of breeder chickens were accidentally exposed to Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS), presumably from a newly introduced group of leghorn-type pullets. The experimental strains subsequently became infected and were diagnosed positive for MG and MS by the serum plate agglutination (SPA) test and confirmed by the hemagglutination inhibition (HI) test and the polymerase chain reaction (PCR) of tracheal swabs. Treatment with 10 mg/kg enrofloxacin via drinking water for 14 days was elected. Before and after initiation of treatment, MG and MS were monitored for changes by SPA, HI, PCR, and culture, with sampling intervals ranging from 1 wk to 7 wk. MG and MS SPA, HI, PCR, and culture were performed at each sampling period, with the exception of weeks 1.0 and 6.5. Week 1.0 included SPA and His for MG and MS. Week 6.5 included PCR and culture for MG and MS. The MG and MS SPA results were positive throughout the 29-wk trial period. MG HI titers declined until the last sampling, whereas the MS HI titers did not decline significantly. PCR for MG yielded only one positive result, which occurred before treatment. MS PCR remained positive throughout the trial period. MG was never isolated from any sample; however, one MS organism was isolated during treatment. The treatment regimen was effective for MG on the basis of PCR results. Treatment with enrofloxacin did not eliminate SPA reactions during the 29-wk trial period. MG HI titers remained in the suspicious range throughout the remainder of the trial period. Four weeks after the treatment ended, MG HIs were reduced by approximately 40%, with MS HIs remaining high throughout the 29-wk period. PCR appeared to be a sensitive and specific test on the basis of correlation with HIs. On the basis of the isolation of MS during treatment and continued subsequent PCR positive reactions, the treatment for MS with enrofloxacin was not as efficacious as for MG. PMID- 11417842 TI - Infraorbital sinusitis associated with Pasteurella multocida in pen-raised ring necked pheasants. AB - Pasteurella multocida, somatic serotype 6, was isolated from the infraorbital sinuses of 8-wk-old ring-necked pheasants with severe sinusitis. In addition, Escherichia coli, Pasteurella haemolytica-like bacteria, Mycoplasma gallinaceum, and Mycoplasma glycophilum were also isolated from some of the sinuses. Clinical signs appeared 3 days after placement on the grow-out ranch. The sinusitis consisted of severe unilateral or bilateral distention of the sinuses by mucoid to caseous exudate. Mortality and morbidity were low. Birds responded to treatment with tetracycline after proper medication procedures. The source of the infection was not determined, though possible sources include the brood ranch, wild animals, or wild waterfowl. PMID- 11417843 TI - Hypoxia induces apoptosis by caspase activation accompanying cytochrome C release from mitochondria in MC3T3E1 osteoblasts. p38 MAPK is related in hypoxia-induced apoptosis. AB - The aim of this study is to elucidate the possible mechanism of apoptosis in response to hypoxia in MC3T3E1 osteoblasts. MC3T3E1 osteoblasts under hypoxic conditions (2% oxygen) resulted in apoptosis in a time-dependent manner estimated by DNA fragmentation assay and nuclear morphologystained with fluorescent dye, Hoechst 33258. Pretreatment with Z-VAD-FMK, a pan-caspase inhibitor, or Z-DEVD CHO, a specific caspase-3 inhibitor, completely suppressed the DNA ladder in response to hypoxia. An increase in caspase-3-like protease (DEVDase) activity was observed during apoptosis, but no caspase- activity (YVADase) was detected. To confirm what caspases are involved in apoptosis, western blot analysis was performed using anti-caspase-3 or -6 antibody. The 10-kDa protein, corresponding to the active products of caspase-3 and the 10-kDA protein of the active protein of caspase-6 were generated in hypoxia-challenged cells in which processing of the full length form of caspase-3 and -6 was evident. With a time course similar to this caspase-3 and -6 activation was evident, hypoxic stress caused the cleavage of lamin A, typical of caspase-6 activity. In addition, the stress elicited the release of cytochrome c into the cytosol during apoptosis. Furthermore, we have observed that pre-treatment with SB203580, a selective p38 MAP kinase (p38 MAPK) inhibitor, attenuated the hypoxia-induced apoptosis. The addition of SB203580 suppressed caspase-3 and -6-like protease activity by hypoxia up to 50%. In contrast, PD98059 had no effect on the hypoxia-induced apoptosis. To confirm the involvement of MAP kinase, JNK/SAPK, ERK, or p38 kinase assay was performed. Although p38 MAPK was activated in response to hypoxic treatment, the other MAP kinase -JNK/SAPK or ERK- was not or modestly activated. These results suggest that p38 MAPK positively regulates hypoxia-induced apoptosis in MC3T3E1 osteoblasts. PMID- 11417844 TI - Splenectomy and sepsis: the role of the spleen in the immune-mediated bacterial clearance. AB - Over the past few years, many observations of overwhelming post splenectomy bacterial infections have been reported. Streptococcus pneumoniae is the aetiologic agent in about 80% of cases, but also gram-negative bacteria are involved in the development of fatal infections in splenectomized patients. Functionally, the spleen plays a fundamental role in bacterial clearance either by antibody response or macrophage bactericidal capacity. At the same time, there is evidence that the spleen also contributes to bacterial endotoxin detoxification. Finally, the mechanisms responsible for gram-positive and gram negative sepsis in the splenectomized host and possible therapeutical approaches able to neutralize bacterial products endowed with noxious effects are discussed. PMID- 11417845 TI - The peripheral blood leukocyte phenotype in patients with breast cancer: effect of doxorubicin/paclitaxel combination chemotherapy. AB - Presence of functional immune system is critical for any attempt aimed at improving survival of breast cancer patients by strategies based on immune system manipulation. We evaluated by flow cytometry the phenotype of peripheral blood leukocyte of 43 breast cancer patients. In 11 patients, the phenotype was evaluated before and during the chemotherapy by combination of doxorubicin and paclitaxel (AT). Compared with controls breast cancer patients had significantly higher relative and absolute numbers of CD3 HLA-DR+, CD3+CD69+ and CD14+CD16+, and significantly lower percentages of CD3 and CD8+CD28+ cells. After one cycle of AT, the absolute numbers of CD3 , CD3+CD4+, CD3+CD8+ and CD8+CD28+ cells increased significantly. Present data show a presence of T-cell activation in breast cancer patients. Administration of AT may lead to an increase in functional T-cells in peripheral blood, indicating a potential for combining chemotherapy with immunotherapy in the treatment of breast cancer patients. PMID- 11417846 TI - Kunbi-Boshin-Hangam-Tang stimulates nitic oxide production through activation of nuclear factor-kappaB. AB - The objective of the currently study was to determine the effect of Kunbi-Boshin Hangam-Tang (KBH-Tang) on the production of nitric oxide (NO). Stimulation of RAW 264.7 cells with KBH-Tang after the treatment of recombinant interferon-gamma (rIFN-gamma) resulted in increased NO synthesis. KBH-Tang partially increased NO synthesis by itself. When KBH-Tang was used in combination with rIFN-gamma, there was a marked cooperative induction of NO synthesis in a dose-dependent manner. This increase in NO synthesis was reflected as increased amount of inducible NO synthase (iNOS) protein. NO production was inhibited by NG-monomethyl-L-arginine (NGMMA). Furthermore, activation of nuclear factor (NF)-kappaB was increased by KBH-Tang. These results suggest that KBH-Tang may stimulate the NO production through the activation of the NF-kappaB. PMID- 11417847 TI - Signal transduction of nitric oxide donor-induced protection in hydrogen peroxide mediated apoptosis in H9C2 cardiomyoblasts. AB - Nitric oxide (NO) attenuates hydrogen peroxide (H2O2)-mediated injury to H9C2 cardiomyoblasts. To examine the role of nitric oxide, cultured H9C2 cardiomyoblasts were treated with H2O2 for 2 h in the presence or absence of the NO donor, diethylamine nitric oxide (DEANO). DEANO (30 microM) attenuated H2O2 induced apoptosis in H9C2 cells. H2O2-exposed H9C2 cells resulted in apoptosis in a time-dependent manner estimated by DNA fragmentation assay, nuclear morphology stained with fluorescent dye, Hoechst 33258 and Annexin V staining. Pretreatment with z-VAD-FMK, a pancaspase inhibitor, or z-DEVD-CHO, a specific caspase-3 inhibitor, completely suppressed the DNA ladder in response to H2O2. An increase in caspase-3-like protease (DEVDase) activity was observed during apoptosis, but no caspase-1 activity (YVADase) was detected. Treatment of H9C2 cells with 100 microM H2O2, resulted in a strong activation of JNK/SAPK. However, the activation of JNK/ SAPK was clearly attenuated by 30 microM DEANO. Furthermore, the dominant negative JNK and SEK1-expressing cells displayed a marked decrease in a number of apoptotic cells. This inhibition of JNK1 in the system is involved in the protection of H2O2-induced apoptosis in H9C2 cardiomyoblasts. PMID- 11417848 TI - Cyclosporin A in patients affected by chronic idiopathic urticaria: a therapeutic alternative. AB - Chronic Idiopathic Urticaria (CIU) is a cutaneous disorder for which there is no identifiable specific etiologic agent. Some recent evidences suggest that CIU might be an autoimmune disease. We analyzed immunological features occurring in CIU and evaluated effectiveness and tolerance of Cyclosporin A (CsA) treatment in patients unresponsive to antihistaminic treatment. Twenty patients with CIU were recruited after a selective diagnostic protocol and were divided into two groups. CsA was prescribed for group 1 and Prednisone for group 2 as control, for 8 weeks. Before and after the therapy we performed on all patients immunological studies. For all patients symptoms disappeared after a few days of therapy. Before therapy all patients showed activated B cells (CD19+CD23+ cells) and among B CD19+ cells, about 20% were CD5+ (cells that synthesize natural autoantibodies). After treatment with Prednisone in group 2, a significant reduction of CD4+ lymphocytes (p = 0,01) was observed. Our findings might support the CIU autoimmune pathogenetic hypothesis. The clinical remission in the CsA treated group confirmed the therapeutic effectiveness of this therapy in antihistaminic unresponsive CIU and, at dosage used, side effects were rare, mild and reversible. Thus, CsA might be a good therapeutic alternative in CIU patients unresponsive to conventional treatments. PMID- 11417849 TI - Adjuvant effect of Pluchea quitoc extract on the resistance of tumor-bearing mice by modulation of the host hematopoietic response. AB - Progressive tumor growth is regularly accompanied by changes in the cellular constituents of the immune system. Evidence suggests that soluble factors generated during tumor growth can affect the amount of granulocyte-macrophage progenitors. In vitro colony growth of progenitor cells may be an early indicator of the cellular changes associated with tumor growth. Pluchea quitoc has been previously found to modulate the hematopoietic response during bacterial infection. This study was designed to investigate the effects of P. quitoc on the growth and differentiation of bone marrow granulocyte-macrophage progenitor cells (CFU-GM) in Ehrlich ascites tumor-bearing mice. In contrast to the myelosuppression developed in the tumor-bearing animals, treatment with P. quitoc ethanolic extract (250, 500 or 1000 mg/kg) for 3 consecutive days after tumor challenge reversibly stimulated myelopoiesis, restoring the number of CFU-GM to normal. This same dose-schedule also increased colony formation in normal mice as compared to controls. In addi tion, P. quitoc significantly enhanced survival of tumor-bearing mice. These results suggest an immunoregulatory role for P. quitoc in counteracting the tumor-induced myelopoietic suppression as well as usefulness as adjuvant treatment of cancer. PMID- 11417850 TI - Disodium cromoglycate inhibits production of immunoglobulin E. AB - Disodium cromoglycate (DSCG) has been shown to inhibit the release of mediators from mast cells. In the present study, the effect of DSCG on active anaphylactic reaction was studied in mice. DSCG dose-dependently inhibited the active systemic anaphylactic reaction and serum immunoglobulin (Ig)E production induced by immunization with ovalbumin, Bordetella pertussis toxin and aluminum hydroxide gel. DSCG strongly inhibited IL-4-dependent IgE production by lipopolysaccharide stimulated murine whole spleen cells. In the case of U266 human IgE-bearing B cells, DSCG also showed an inhibitory effect on the IgE production. These results suggest that DSCG has an anti-anaphylactic activity by inhibition of IgE production from B cells. PMID- 11417851 TI - Danchunhwan water extract prevents apoptotic death by peroxynitrite and nitric oxide in human dopaminergic neuroblastoma SH-SY5Y cells. AB - In the present study, the protective effects of Danchunhwan on the cytotoxicity by peroxynitrite and nitric oxide (NO) were investigated in human dopaminergic neuroblastoma SH-SYSY cells. Danchunhwan has been used to treat infarction and cerebrovascular diseases in Oriental medicine for centuries. Cells were pretreated with Danchunhwan and exposed to sodium nitroprusside (SNP), an NO donor, and 3-morpholinosydnonimine (SIN-1) which simultaneously generates NO and superoxide, thus possibly forming peroxynitrite. Exposure of cells to SIN-1 for 24 hr induced 75% of apoptotic cell death, as evaluated by ladder-pattern fragmentation of genomic DNA and characteristic of apoptosis using 4', 6 diamidino-2-phenylinol (DAPI). However, pretreatment of SH-SY5Y cells with Danchunhwan inhibited the apoptotic cell death in a dose-dependent manner. Even though Danchunhwan was washed out after preincubation for 12 hr, cells were still remained to be resistant against cytotoxicity of SIN-1. It also inhibited SIN-1 induced activation of caspase 3-like protease in a dose-dependent fashion. Furthermore, Danchunhwan recovered the levels of intracellular antioxidant system, reduced glutathione (GSH) (83%), which was decreased by the addition of SIN-1 (63%). Taken together, we suggest that Danchunhwan protects human neuroblastoma SH-SY5Y cells from apoptotic death by free radicals including peroxynitrite and NO via generation of antioxidant, GSH. PMID- 11417852 TI - Reduction of acute inflammation in rats by diazepam: role of peripheral benzodiazepine receptors and corticosterone. AB - Carrageenin causes a reproducible inflammatory reaction and remains the standard irritant for examining acute inflammation and anti-inflammatory drugs. High doses of diazepam (10.0-20.0 mg/Kg) were shown to reduce the volume of acute inflammatory paw edema in rats as a response to carrageenin administration. The present experiment was undertaken to investigate the possible roles of peripheral type benzodiazepine receptors (PBRs) and corticosterone on the anti-inflammatory effects of diazepam. Five experiments were conducted to assess the effects of a single dose (10.0 mg/Kg) of diazepam on carrageenin-induced paw edema (CIPE), pleurisy and increase in vascular permeability in rats. Results showed that: 1. diazepam or Ro5-4864 (a PBR agonist) treatment reduced CIPE values; 2. prior treatment with PK11195 (a non-benzodiazepine PBR antagonist) suppressed the effects of either diazepam or Ro5-4864 on CIPE; 3. diazepam reduced the volume of the pleural exudate in carrageenin-injected rats, as well as its leukocyte count; 4. diazepam treatment reduced the magnitude of the increase in vascular permeability caused by carrageenin; 5. adrenalectomy suppressed the effects of diazepam on CIPE; and 6. diazepam treatment increased the serum concentration of corticosterone. These results suggest a relevant role of PBR and corticosterone on diazepam-induced changes in inflammation. They are discussed in the light of a possible activation of mitochondrial PBRs within the adrenal gland cells by diazepam, thereby increasing the serum levels of corticosterone and thus reducing CIPE. PMID- 11417853 TI - Immunomodulation in progeny from thymectomized primiparous mice exposed to benzo(a)pyrene during mid-pregnancy. AB - Previous studies have shown that Benzo(a)pyrene (B(a)P3) given to non thymectomized (NTX) female mice alters expression of T cell subsets and suppresses cell mediated immunity (CMI) and humoral immunity (HI) in the progeny. Thus, maternal exposure to B(a)P may influence changes in progeny immune status. To understand how maternal cellular and humoral factors influence embryonic development of progeny immunity, adult female mice were thymectomized (TX) at 6 weeks, mated and injected with 150 microg B(a)P)/g body weight at 12 days of pregnancy. After B(a)P exposure, the following studies were performed: (A) Maternal reproductive capacity and survival rate of progeny; (B) Detection of T cells in progeny thymus; (C) Functional characteristics of progeny thymus or spleen. Maternal thymectomy and B(a)P exposure reduced average litter size by 40%. Serological sensitivity of thymus cells with anti-Thyl + complement occurred at a higher dilution of mAb in progeny from TX mothers exposed to B(a)P, suggesting that B(a)P-thymectomy led to increased sensitivity of developing thymocytes to mAb plus complement. Progeny from TX mothers exposed to B(a)P showed enhanced thymic CMI, but suppressed splenic CMI and HI. Thus, thymectomy prevents CMI immunosuppression by B(a)P, while HI is still suppressed. These results indicate that the maternal thymus is necessary for incurring the effect of B(a)P on progeny CMI. PMID- 11417854 TI - Enhancement of chicken macrophage phagocytic function and nitrite production by dietary Spirulina platensis. AB - The effects of dietary Spirulina platensis on chicken macrophage phagocytic function and nitrite production were examined. Day old broiler (meat-type) chicks were randomly assigned to various pens of electrically heated wire batteries. Dietary treatment groups included a basal diet with no dietary Spirulina added, and three additional groups with 0.5, 1.0 and 2.0% dietary Spirulina. Feed and water were provided for ad libitum consumption from one day of age. Sephadex elicited macrophages were harvested at 14, 35 and 42 days of age. Phagocytosis assay was performed by co-incubating sheep red blood cells (SRBC) with the adherent macrophage monolayers. For nitrite quantification, macrophage cultures from various dietary treatment groups were stimulated in the presence or absence of 1 microg/mL of Escherichia coli lipopolysaccharide. These culture supernatant fractions were then tested for nitrite levels using the Greiss reagent technique. All Spirulina dietary group macrophages exhibited an enhanced phagocytic activity in terms of overall phagocytic percentage (range = 28 to 39% versus 24 to 25% in the basal group) and the average number of SRBC per phagocytic macrophage (range = 2.2 to 3.6 versus 1.8 to 2.5 in the basal group). This increase was linear with each incremental increase of dietary Spirulina. While LPS-induced nitrite levels in macrophages from basal diet group ranged from 60 to 278 microM over the three developmental ages, these levels in all Spirulina dietary groups were significantly higher (0.5% group range = 198 to 457 microM; 1.0% group range = 161 to 359 microM and 2.0% group range = 204 to 420 microM. These data clearly show that Spirulina platensis feeding upregulates macrophage phagocytic as well as metabolic pathways leading to increased nitric oxide synthase activity. These findings therefore imply that Spirulina platensis may enhance the functions of mononuclear phagocytic system thereby increasing the disease resistance potential in chickens. PMID- 11417855 TI - Some inflammation-related parameters in patients following normo- and hypothermic cardio-pulmonary bypass. AB - AIMS: One of the complications of Cardio-Pulmonary Bypass is the Systemic Inflammatory Response Syndrome. Cardio-Pulmonary Bypass can be performed under either normothermic or hypothermic conditions. The aim of this study was to compare some inflammation-related parameters of patients following normothermic and hypothermic bypass. Moreover, attempts were undertaken to detect endotoxin, an inflammatory agent that has been implicated in the Systemic Inflammatory Response Syndrome, in the serum of patients. Levels of serum anti-endotoxin antibodies were estimated since they have been reported to negate the effect of endotoxin in the inflammatory syndrome. METHODS AND RESULTS: Seventeen normothermic and 20 hypothermic cases were studied. Blood specimens were collected pre-, off- and post-bypass. Pertinent clinical and surgical data were collected. Hematological parameters (leukocyte, neutrophil and platelet counts) and liver function tests were determined by standard procedures. Endotoxin was determined by the Limulus Lysate Assay and anti-endotoxin antibodies by an enzyme immunoassay. Complement (C3 and C4) levels were determined by radial immunodiffusion. There were increases in leukocyte and neutrophil, and a decline in platelet numbers in both groups of patients. There was a decline in C3 and C4 levels in both groups of patients. Endotoxin was not detected in sera, and anti endotoxin antibody levels were similar, in both groups of patients. CONCLUSION: There were no significant differences in most of the altered inflammation-related parameters between the two groups of patients. Some of the findings might be partly due to hemo-dilution. The hydrophobic nature of endotoxin among other factors, might have hindered its detection in serum. PMID- 11417856 TI - A syndrome characterized by psychiatric disorders, recurrent mucosal infections and natural immunity deficits: clinical approach. AB - The authors summarize their own previous work on the identification of a subset of patients characterized by psychiatric disorders, recurrency of mucosal infections and impaired natural immunity. The diagnostic approach to these patients based on the close collaboration between infectivologists, immunologists and psychiatrists is described with the aim to find out combined treatments for the amelioration of clinical manifestations. PMID- 11417857 TI - T cell-mediated antigen presentation: a potential mechanism of infectious tolerance. AB - Differentiation of the T cell repertoire and the physiology of T cell-mediated antigen presentation are reviewed in relation to mechanisms of self-tolerance. Recent research has indicated that T cell development is a continual process that optimizes partial recognition of self as a homeostatic set-point. Specific T cell antigen recognition of partial agonists is intrinsically linked to expression of class II MHC glycoproteins on T cells. Even ligands that act as TCR antagonists in IL-2 production assays have sufficient agonistic strength to induce expression of class II MHC glycoproteins on T cells. Thus, the intrinsic self-reactivity of the T cell repertoire may promote T-APC activity in vivo and may explain why thymic and peripheral T cells express low but significant levels of class II MHC glycoproteins. T-APC activity induces extensive apoptosis among responder T cells, causes desensitization among surviving responders, and has been implicated in the adoptive transfer of tolerance in the Lewis rat model of experimental autoimmune encephalomyelitis. Overall, these findings support a relationship between the partial recognition of self MHC ligands, expression of class II MHC glycoproteins on mature peripheral T cells, tolerogenic T cell-mediated antigen presentation, and desensitization of pathogenic self-reactive T cells. PMID- 11417858 TI - RAG1 and RAG2 in V(D)J recombination and transposition. AB - RAG1 and RAG2 are the key components of the V(D)J recombinase machinery that catalyses the somatic gene rearrangements of antigen receptor genes during lymphocyte development. In the first step of V(D)J recombination--DNA cleavage- the RAG proteins act together as an endonuclease to excise the DNA between two individual gene segments. They are also thought to be involved in the subsequent DNA joining step. In vitro, the RAG proteins catalyze the integration of the excised DNA element into target DNA completing a process similar to bacterial transposition. In vivo, this reaction is suppressed by an unknown mechanism. The individual roles of RAG1 and RAG2 in V(D)J recombination and transposition reactions are discussed based on mutation analyses and structure predictions. PMID- 11417861 TI - Individuality of Ag-selected and preimmune TCR repertoires. AB - We have combined flow cytometry and single-cell PCR to characterize the TCRBV repertoires selected by individual mice in a model CD8 response against a defined peptide/MHC complex (CW3 170-1 79/Kd). Ourresults established thatdifferent mice select individually distinct yet structurally similar CW3-specific repertoires. Repertoire selection appears to be flexible depending on the immunizing cell dose. Using a single-donor, matched-pair-recipient adoptive transfer strategy, we demonstrated that the CW3-specific TCR repertoires of normal mice are already distinct at the preimmune level. We combine our data with computer simulations to test models for the composition of an Ag-specific preimmune repertoire and its selection during an immune response. PMID- 11417862 TI - Neutrophil oscillations: temporal and spatiotemporal aspects of cell behavior. AB - Neutrophil activation is an essential event in inflammatory responses. How cells coordinate, integrate, manage, and distribute information on physiologically relevant timescales are not well understood. Although neutrophil oscillators have been known for many years, their biological roles have not been identified. We suggest that intracellular oscillators (such as NAD(P)H, pH, calcium, and so on) account for functional oscillations (e.g., superoxide and NO production, cytolytic marker release, pericellular proteolysis, and actin assembly). In addition to these well-known temporal oscillations, we have recently discovered self-organized traveling chemical waves in neutrophils; these waves respond to extracellular signals and have distinct origins that coincide with a cell's uropod, lamellipodium, or adherence site. The fundamental physico-chemical features of cell chemistry will have an increasing role in our understanding of leukocyte function. PMID- 11417860 TI - Cytokines in posterior uveitis: an update. AB - T helper (Th) lymphocytes differentiate into two distinct subsets--Th1 and Th2- as defined by functional abilities and cytokine profiles. The functional differences between Th subsets are explained primarily through the activities of the cytokines they secrete. Interferon-gamma (IFN-gamma) is the signature cytokine of Th1 cells, which also produce interleukin-2 (IL-2) and tumor necrosis factor-beta (TNF-beta). IL-4 is the corresponding signature cytokine of Th2 cells, which also secrete IL-5, IL-6, IL-9, and IL-13. Recently, a few transcription factors have been identified that not only control the expression of cytokines of a particular type but also repress cytokines of other types. Human Th1 and Th2 cells not only produce a different set of cytokines but also exhibit distinct functional properties and the preferential expression of some activation markers. Pathophysiologically, the two subsets have been found to be mutually antagonistic. Various Th1 and Th2 cytokines appear to play an important role in the etiopathogenesis of posterior uveitis and its animal model, experimental autoimmune uveitis (EAU). The exact contribution of these mediators to uveitis remains to be defined. Recent studies suggest that a shift from Th1- to Th2-dominated response could be of therapeutic benefit. This review evaluates various studies in which uveitopathogenic and therapeutic potentials of various Th1 and Th2 cytokines have been investigated. PMID- 11417859 TI - IL-6-like cytokines and cancer cachexia: consequences of chronic inflammation. AB - An estimated 30% of cancer deaths are attributed to cachexia and its consequences. Cachexia (wasting syndrome) is the hypercatabolism of the body's carbon sources, proteins and lipids, for conversion into energy. It is induced by a variety of pathological conditions, including cancer. Among the inflammatory responses to cancer is the synthesis of cytokines, including IL-6 and related cytokines. These cytokines have been found to induce cachexia by altering metabolism of lipids and proteins. IL-6-like cytokines have been found to inhibit lipid biosynthesis by adipocytes, which increased the rate of lipid catabolism. Others have described the atrophy and increased catabolism of muscle protein due to IL-6. A cytokine closely-related to IL-6 is leptin, which plays a major role in lipid metabolism under normal conditions. The role of leptin in pathological conditions such as cancer cachexia has not yet been fully elucidated. Detailed mechanistic information about the induction of cancer cachexia by IL-6-like cytokines requires more research. PMID- 11417863 TI - High-performance liquid chromatographic assay with fluorescence detection for the determination of cephaeline and emetine in human plasma and urine. AB - A high-performance liquid chromatographic assay method for the quantitation of ipecac alkaloids (cephaeline and emetine) in human plasma and urine is described. Human plasma or urine was extracted with diethylether under alkaline conditions following the addition of an internal standard. Concentrations of alkaloids and internal standard were determined by octadecylsilica chromatographic separation (Symmetry C18 columns, plasma analysis; 15 cmx4.6 mm I.D., 5 microm particle size, urine analysis; 7.5 cmx4.6 mm I.D., 5 microm particle size). The mobile phase consisted of buffer (20 mmol/l 1-heptanesulfonic acid sodium salt, adjusted to pH 4.0 with acetic acid)-methanol (51:49, v/v). Eluate fluorescence was monitored at 285/316 nm. The lowest quantitation limits of cephaeline and emetine were 1 and 2.5 ng/ml, respectively, in plasma, and 5 ng/ml in urine. Intra- and inter-day relative standard deviations were below 15%. The assay is sensitive, specific and applicable to pharmacokinetic studies in humans. PMID- 11417864 TI - Gas chromatographic-mass spectrometric assay for rocuronium with potential for quantifying its metabolite, 17-desacetylrocuronium, in human plasma. AB - A rapid, sensitive and selective method has been developed for the quantification of plasma concentrations of neuromuscular blocking drug, rocuronium, using gas chromatography with mass spectrometric detection. 3-Desacetylvecuronium served as the internal standard. The method involved iodide ion pair formation and a single step liquid-liquid extraction with dicholoromethane. This method also permits simultaneous determination of its putative metabolite, 17-desacetylrocuronium, although the high detection limit for the metabolite limits the practical application of this method in pharmacokinetic study of the metabolite. The extraction efficiency was approximately 75% for rocuronium and approximately 50% for 17-desacetylrocuronium. The limit of quantification was 26 ng/ml for rocuronium and 870 ng/ml for its metabolite. The assay was used successfully in a patient undergoing liver transplantation and receiving rocuronium as a constant rate infusion and in a patient undergoing general elective surgery receiving the drug as an intravenous bolus. This assay is a time-saving alternative to published gas or liquid chromatographic methods for assaying rocuronium. PMID- 11417865 TI - Determination of fendiline and gallopamil by capillary isotachophoresis. AB - Capillary isotachophoresis was applied for the determination of fendiline and gallopamil--calcium antagonists--in serum. The cationic electrolyte system containing Na+ with acetic acid as a counter constituent was used as a leading electrolyte with the pH 4.7 and the terminating electrolyte was beta-alanine. Most of the proteins were precipitated with methanol, ethanol and dimethylketone. The lowest limits of quantitation were obtained for the pretreatment of serum with methanol. The recoveries of both compounds varied from 91.3 to 97.5%. The relative standard deviations varied from 0.6 to 7.7%. PMID- 11417866 TI - Comparison of the efficiences of enzymatic and chemical hydrolysis of (nortestosterone and diethylstilboestrol) glucuronides in bovine urine. AB - Residues of 19-nortestosterone (19-NT) and diethylstilboestrol (DES) are excreted in bovine urine, mainly conjugated to glucuronic acid. Prior to quantification, urine must be deconjugated, which is commonly performed by enzymatic or chemical hydrolysis. The efficiencies of two enzymatic and two chemical deconjugation methods were studied. The range of efficiencies obtained for DES were 51.8% (beta glucuronidase, incubation at 37 degrees C overnight) and 2.7% (methanolic HCl), respectively. Similarly, efficiencies for NT ranged from 43.1% (beta glucuronidase, incubation at 55 degrees C for 2 h) to 12.7% (methanolic HCl). The results highlight that within control laboratories significant underestimation of drug residue content in samples may occur, due to poor deconjugation. PMID- 11417867 TI - High-performance liquid chromatography with chemiluminescence detection of penbutolol and its hydroxylated metabolite in rat plasma. AB - This paper describes a new method of high-performance liquid chromatography with chemiluminescence detection for the analysis of penbutolol (PB) and its main metabolite, 4-hydroxy penbutolol (4-OH PB) in rat plasma. 4-Dimethylaminosulfonyl 7-(N-chloroformylmethyl-N-methyl) amino-2,1,3-benzoxadiazole (DBD-COCl) was used as a fluorogenic labeling reagent. A mixture of hydrogen peroxide and bis[4-nitro 2-(3,6,9-trioxadecyloxycarbonyl)phenyl]oxalate (TDPO) in acetonitrile was used as a post-column chemiluminogenic reagent. The derivatives of PB and 4-OH PB with DBD-COCI were separated by isocratic effluent with 0.01 M imidazole buffer (pH 7.0)-acetonitrile within 10 min. The detection limits of the proposed method for PB and 4-OH PB were 9.9 and 15 fmol on column, respectively. After intravenous administration of PB in rats, its plasma concentration profiles of PB and 4-OH PB were determined by the proposed method. PB was demonstrated to be rapidly metabolized to 4-OH PB at the same rate as cardiac output. PMID- 11417868 TI - Amino acid modifications in canine, equine and porcine pituitary growth hormones, identified by peptide-mass mapping. AB - Modified amino acid residues in porcine, canine and equine growth hormones purified from pituitary glands were characterised by tryptic mapping and high performance liquid chromatography with on-line coupled electrospray ionisation mass spectrometry (HPLC-ESI-MS) detection. Hormones from all three species showed the same changes. Conversion of Asp128 to iso-Asp128 was a component of native hormones, while deamidation of Asn12 and Asn98 to Asp and iso-Asp, oxidation of Met4, and cyclisation to the pyroglutamyl derivative of Gln139, probably occurred in vitro, during isolation, storage or hydrolysis. Porcine and canine hormones had indistinguishable protein fingerprints, confirming the assumption, based on their cDNA sequences, that their mature primary structures are identical. PMID- 11417869 TI - Determination of phytic acid by gas chromatography-mass spectroscopy: application to biological samples. AB - A GC-MS method is reported for the determination of phytic acid based on purification by anion-exchange chromatography, enzymatic hydrolysis of phytic acid to myo-inositol and derivation to trimethylsilyl derivative, with scyllo inositol as an internal standard. Analytical features of the method are: limit of detection 9 microg l(-1) phytic acid, linear working range 18-500 microg l(-1) phytic acid, and coefficient of variation 1.9%. The method has been successfully applied to a variety of biological samples: various rat organs (kidney, liver, brain and bone), human plasma and urine and kidney stones. A comparative study of sample treatments, including deproteization, lipid extraction and the presence of a chelator, is also reported. Phytic acid amounts found in rat organs ranged from 1.07 g kg(-1) for bone to 32.0 g kg(-1) for brain. Phytic acid in human plasma was of the order of 0.14 mg l(-1). In kidney stones, phytic acid was found in calcium containing stones. PMID- 11417870 TI - Separation and determination of liposomal and non-liposomal daunorubicin from the plasma of patients treated with Daunoxome. AB - Several liposomal formulations of anthracyclines have been developed recently and are currently used in the clinical setting. We describe a technique of separation and quantification of the liposomal and non-liposomal forms of daunorubicin in the plasma of patients treated with DaunoXome, a liposomal formulation of daunorubicin. The method we propose is based upon the property of liposomes to cross reversed-phase C18 silicagel cartridges without being retained, while non liposomal drug is retained on the stationary phase and is eluted with methanol. Extraction of liposomal and non-liposomal daunorubicin from plasma, therefore, is performed in two steps. This technique is rapid, can be automated in order to handle large series of samples, and the plasma can be frozen after sampling by addition of glycerol. The recovery of liposomal daunorubicin as well as the precision, linearity and accuracy of the technique appear satisfactory for pharmacokinetic purposes. PMID- 11417871 TI - Heparin column analysis of serum type 5 tartrate-resistant acid phosphatase isoforms. AB - The objective of the present study was to develop a specific method for the separation of tartrate-resistant acid phosphatase (TRAP) derived exclusively from osteoclasts. Heparin column-bound TRAP in human serum was separated into three peaks of TRAP activity when eluted with a linear gradient of sodium chloride. The last peak corresponded to TRAP 5b which was first named according to its electrophoretic mobility [Clin. Chem. 24 (1978) 309] and was considered to be derived from osteoclasts [J. Bone Miner. Res. 13 (1998) 683]. The second peak was found to be TRAP 5a. The height of the last peak varied from age to age. PMID- 11417872 TI - Rapid and sensitive step gradient assays of glutamate, glycine, taurine and gamma aminobutyric acid by high-performance liquid chromatography-fluorescence detection with o-phthalaldehyde-mercaptoethanol derivatization with an emphasis on microdialysis samples. AB - We developed a rapid step-gradient HPLC method for determination of glutamate, glycine and taurine, and a separate method for determination of gamma aminobutyric acid (GABA) in striatal microdialysates. The amino acids were pre column derivatized with o-phthalaldehyde-2-mercaptoethanol by using an automated refrigerated autoinjector. Separation of the amino acids was established with a non-porous ODS-II HPLC column, late-eluting substances were washed out with a one step low-pressure gradient. Concentrations of the amino acids were determined with a fixed-wavelength fluorescence detector. The detection limit for GABA was 80 fmol in a 15 microl sample, detection limits for glutamate, glycine and taurine were not determined because their concentrations in striatal perfusates were far above their detection limits. Total analysis time was less than 12 min, including the wash-out step. The methods described are relatively simple, sensitive, inexpensive, and fast enough to keep up with the microdialysis sampling. PMID- 11417873 TI - On the isolation of polychlorinated dibenzo-p-dioxins and furans from serum samples using immunoaffinity chromatography prior to high-resolution gas chromatography-mass spectrometry. AB - Immunoaffinity chromatography (IAC) for the purification of polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs) from biological samples was explored as a means to simplify the cleanup procedure and thereby decrease the time and cost of dioxin analysis. A monoclonal antibody (DD3) was used to produce IAC columns and to isolate the PCDD/Fs from serum. Native and 13C-labeled PCDD/Fs were spiked at the ppq to ppt range into serum. Quantitation of the PCDD/Fs was performed by a standard dioxin analytical method, i.e. high-resolution gas chromatography-mass spectrometry (GC-MS), which was easily compatible with IAC. Five of the most toxic PCDD/Fs consistently showed acceptable recoveries (>25%) and were reliably quantitated. The congeners specifically recovered by this method represent almost 80% of the toxic equivalency of dioxins and furans present in the serum samples. Dioxin-like polychlorinated biphenyls (PCBs) were not recognized by this antibody column. Compared to conventional dioxin cleanup methods, IAC decreased solvent usage by 1.5 l/sample and took only 2 h to process a sample for analysis. PMID- 11417874 TI - Quantification of nicotine, chlorpyrifos and their metabolites in rat plasma and urine using high-performance liquid chromatography. AB - This study describes a high-performance liquid chromatographic method for the separation and quantification of nicotine, its metabolites nornicotine and cotinine, the insecticide chlorpyrifos (O,O-diethyl-O[3,5,6-trichloro-2 pyridinyl]phosphorothioate), and its metabolites chlorpyrifos-oxon (O,O-diethyl O[3,5,6-trichloro-2-pyridinyl]phosphate), and TCP (3,5,6-trichloro-2-pyridinol) in rat plasma and urine. The compounds were separated using gradient mobile phase of methanol, acetonitrile and water (pH 3.20) at a flow-rate of 0.8 ml/min in a period of 17 min, and gradient UV detection ranging between 260 and 280 nm. The retention times ranged from 3.4 to 16.7 min. The limits of detection were ranged between 20 and 150 ng/ml, while limits of quantitation were 50-200 ng/ml. Average percentage recovery of five spiked plasma samples were 84.7+/-8.3, 78.2+/-7.6, 80.1+/-7.6, 79.0+/-6.4, 74.0+/-7.4, 87.6+/-7.5, and from urine 85.1+/-5.2, 75.9+/ 7.0, 82.1+/-6.1, 79.5+/-6.1, 71.3+/-7.4 and 81.3+/-6.9 for nicotine, nornicotine, cotinine chlorpyrifos, chlorpyrifos-oxon and TCP, respectively. Intra-day accuracy and precision for this method were ranged between 2.2-3.6 and 2.1-2.8%, respectively. The relationship between peak areas and concentration was linear over range between 200 and 2000 ng/ml. This method was applied to analyze the above chemicals and metabolites following combined oral administration in rats. PMID- 11417875 TI - Comparison of ion-pair and amide-based column reversed-phase liquid chromatography for the separation of thiamine-related compounds. AB - Two reversed-phase chromatographic methods for the separation of thiamine and related compounds are compared. The first procedure is based on the ion-pair technique using an octadecylsilica column, while the second uses a new amide based stationary phase, which avoids the need to form ion-pairs, leading to narrower peaks and a simpler mobile phase. Analyses were performed by gradient elution and a photo-diode array was used for detection. Specificity was demonstrated by the retention characteristics, UV spectra and by comparing the peak purity index with commercial standards. The procedures were applied to the determination of thiamine-related compounds in pharmaceutical preparations and urine. No preliminary sample treatment was required. PMID- 11417876 TI - Metabolism of amiodarone. II. High-performance liquid chromatographic assay for mono-N-desethylamiodarone hydroxylation in liver microsomes. AB - Amiodarone (AMI) is a potent antiarrhythmic drug. In vivo and in vitro, AMI is biotransformed to mono-N-desethylamiodarone (MDEA). Recently, it was observed that MDEA was further hydroxylated to n-3'-hydroxybutyl-MDEA (3'OH-MDEA). The performance of a HPLC-UV assay being developed for the quantification of the new compound was investigated. Liver microsomes isolated from rabbit, rat and human biotransformed MDEA to 3'OH-MDEA. Their estimates of Michaelis-Menten parameters were Km=6.39, 25.2, 19.4 microM; Vmax=560, 54, 17.3 pmol/mg protein/min), respectively. Thus, hydroxylase activity in mammals may be the origin of the species dependence observed in the AMI metabolism. PMID- 11417877 TI - Sensitive fluorescent quantitation of myo-inositol 1,2-cyclic phosphate and myo inositol 1-phosphate by high-performance thin-layer chromatography. AB - A non-radioactive micro-assay for the cyclic phosphodiesterase reaction catalyzed by Bacillus cereus phosphatidylinositol-specific phospholipase C is described. The assay involves high-performance thin-layer chromatography on silica gel to resolve the substrate (myo-inositol 1,2-cyclic phosphate) and the product (myo inositol 1-phosphate), followed by detection with a lead tetraacetate-fluorescein stain. The quantitation of these inositol phosphates in sample spots relative to a series of standards is accomplished by analysis of the fluorescent plate image with a commercial phosphoimager and associated software. The experimental considerations for reliable quantitation of inositol monophosphates in the range of 0.1 to 50 nmol are presented. PMID- 11417878 TI - Simultaneous determination of the HIV-protease inhibitors indinavir, amprenavir, ritonavir, saquinavir and nelfinavir in human plasma by reversed-phase high performance liquid chromatography. AB - A rapid, simple and sensitive high-performance liquid chromatographic (HPLC) assay has been developed for the simultaneous quantification of the HIV-protease inhibitors indinavir, amprenavir, ritonavir, saquinavir and nelfinavir in human plasma. The method involved the solid-phase extraction of the five drugs and the internal standard (I.S., verapamil) from 400 microl of human plasma. The HPLC analysis used a reversed-phase C18 analytical column and a mobile phase consisting of a gradient with 15 mM phosphate buffer (pH 5.75)-acetonitrile and UV monitoring. The method was linear over the therapeutic concentration range for the five HIV-protease inhibitors. The accuracy of the method ranged from 98.2 to 106.7% and the precision values ranged from 1.4 to 8.1% for intra-day precision and from 3.1 to 6.4% for the inter-day values. PMID- 11417879 TI - Liquid chromatography-electrospray ionization mass spectrometric analysis of corticosterone in rat plasma using selected ion monitoring. AB - A simple and fast yet highly sensitive and specific method based on HPLC coupled to electrospray ionization mass spectrometry has been developed for the quantitation of corticosterone in rat plasma. After extraction of rat plasma (100 microl) with diethyl ether using 5-pregnen-3beta-ol-20-one-16alpha-carbonitrile (Sigma) as internal standard, HPLC was performed on a short C8 column (Zorbax Eclipse, 50x4.6 mm I.D.) using a steep methanol-water gradient (methanol 54% to 90% in 6 min). Detection was performed on a single quadruple mass spectrometer in selected ion monitoring mode (m/z 369 for corticosterone and 364 for the internal standard). The detection limit of the assay was 9 fmol (3 pg) of corticosterone on column. In vitro data were subjected to curve fitting (cubic, r2=0.9999). Recovery of corticosterone after extraction ranged from 81 to 93%. The relative standard deviations for intra- and inter-assay precision ranged from 0.8 to 3.6% and 5.2 to 12.9%, respectively. Corticosterone did not undergo any appreciable degradation when stored in plasma at -20 degrees C for 2 months. The assay is routinely used in our laboratory to examine corticosterone levels as a marker of stress in rats and may also be used for the determination of 18-hydroxy-11 deoxycorticosterone. PMID- 11417880 TI - Determination of the covalent adducts of the novel anti-cancer agent 5,6 dimethylxanthenone-4-acetic acid in biological samples by high-performance liquid chromatography. AB - The reversed-phase HPLC methods were developed to determinate the covalently bound protein adducts of the novel anti-cancer drug 5,6-dimethylxanthenone-4 acetic acid (DMXAA) via its glucuronides after releasing aglycone by alkaline hydrolysis in human plasma and human serum albumin (HSA). An aliquot of 75 microl of the mixture was injected onto a Spherex C18 column (150x4.6 mm; 5 microm) at a flow-rate of 2.5 ml/min. The mobile phase comprising of acetonitrile:10 mM ammonium acetate buffer (24:76, v/v, pH 5.8) was used in an isocratic condition, and DMXAA was detected by fluorescence. The method was validated with respect to recovery, selectivity, linearity, precision, and accuracy. Calibration curves for DMXAA were constructed in the concentration range of 0.5-40 microM in washed blank human plasma or HSA prior to alkaline hydrolysis. The difference between the theoretical and calculated concentration and the relative standard deviation were less than 10% at all quality control (QC) concentrations. The limit of detection for the covalent adduct in human plasma or HSA is 0.20 microM. The methods presented good accuracy, precision and sensitivity for use in the preclinical and clinical studies. PMID- 11417881 TI - Sensitive gas chromatographic determination of the cyclophosphamide metabolite 2 dechloroethylcyclophosphamide in human plasma. AB - Cyclophosphamide (CP) is one of the most frequently used anticancer agents. It is a prodrug requiring activation before exerting cytotoxicity. CP is deactivated to 2-dechloroethylcyclophosphamide (2-DCECP) with formation of an equimolar amount of chloroacetaldehyde. The aim of this study was to develop and validate a sensitive and simple assay for 2-DCECP in plasma of patients treated with CP. Sample pre-treatment consisted of solid-phase extraction of 500 microl of plasma over OASIS HLB (1 ml) cartridges with trofosfamide as internal standard. Separation and detection of underivatized 2-DCECP was performed with capillary gas chromatography with nitrogen/phosphorous selective detection. Extraction recovery of 2-DCECP exceeded 87%. No interference from endogenous compounds, other metabolites of CP and frequently coadministered drugs was detected. The assay was linear in the range of 5-5000 ng/ml in plasma. Accuracy, within-day and between-day precision were less than 11% for the complete concentration range. In plasma, 2-DCECP was stable for at least 1 month when kept at -70 degrees C. Analysis of samples from patients treated with CP demonstrated the applicability of the assay. In conclusion, a sensitive and simple assay for 2-DCECP in plasma, which meets the current requirements for bioanalytical assays, was developed. PMID- 11417882 TI - Determination of unbound concentration of the novel anti-tumour agent 5,6 dimethylxanthenone-4-acetic acid in human plasma by ultrafiltration followed by high-performance liquid chromatography with fluorimetric detection. AB - The novel anti-tumour agent 5,6-dimethylxanthenone-4-acetic acid (DMXAA) is a highly protein bound drug with narrow therapeutic window. We report a simple HPLC method with fluorimetric detection for the determination of free DMXAA concentration in human plasma. Sample preparation involves the ultrafiltration of plasma by a Centrisart device for 30 min at 2000 g and extraction with acetonitrile: methanol mixture. The method was validated with respect to recovery, selectivity, linearity, precision, and accuracy. Calibration curves for DMXAA were constructed at the concentration range of 0.5-40 microM in blank plasma and phosphate buffer. The difference between the theoretical and calculated concentration and the relative standard deviation were less than 10% at all quality control (QC) concentrations. The HPLC method has been used for the analysis of preclinical studies. PMID- 11417883 TI - Simultaneous quantification of retinol, retinal, and retinoic acid isomers by high-performance liquid chromatography with a simple gradiation. AB - A new method of high-performance liquid chromatography (HPLC) analysis to quantify isomers of retinol, retinal and retinoic acid simultaneously was established. The HPLC system consisted of a silica gel absorption column and a linear gradient with two kinds of solvents containing n-Hexane, 2-propanol, and glacial acetic acid in different ratios. It separated six retinoic acid isomers (13-cis, 9-cis, all-trans, all-trans-4-oxo, 9-cis-4-oxo, 13-cis-4-oxo), three retinal isomers (13-cis-, 9-cis-, and all-trans) and two retinol isomers (13-cis- and all-trans). Human serum samples were subjected to this HPLC analysis and at least, all-trans retinol, 13-cis retinol, and all-trans retinoic acid were detectable. This HPLC system is useful for evaluating retinoic acid formation from retinol via a two-step oxidation pathway. Moreover, it could be applied to monitoring the concentrations of various retinoids, including all-trans retinoic acid in human sera. PMID- 11417884 TI - Fluorometric determination of N-terminal prolyl dipeptides, proline and hydroxyproline in human serum by pre-column high-performance liquid chromatography using 4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsufonyl chloride. AB - A highly sensitive HPLC method for the determination of prolyl dipeptides, Pro and Hyp in serum was developed. After deproteinization of serum and pretreatment with o-phthalaldehyde, the analytes were derivatized with 4-(5,6-dimethoxy-2 phthalimidinyl)-2-methoxyphenylsulfonyl chloride at 70 degrees C for 10 min. The fluorescent derivatives of prolyl dipeptides, Pro and Hyp, were separated on tandem reversed-phase columns by a gradient elution at 55 degrees C and detected by fluorescence measured at 318 nm (excitation) and 392 nm (emission). The detection limits for prolyl dipeptides were 2-5 fmol/injection (S/N=3). Pro-Hyp, Pro-Gly and Pro-Pro were identified as serum prolyl dipeptides. The within-day and between-day relative standard deviations were 1.5-7.9 and 2.4-10.8%, respectively. The recoveries were in the range of 90.8-97.3%. The concentrations of Pro-Hyp, Pro-Gly, Pro-Pro, Pro and Hyp in normal human serum (n = 10) were 0.64+/-0.35, 0.078+/-0.047, 0.022+/-0.016, 177.0+/-43.0 and 11.1+/-3.5 microM, respectively. The concentrations of Pro-Hyp and Pro-Pro in serum of a patient with bone metastases of prostatic cancer were about three times and 50 times, respectively, higher than those in normal human serum. PMID- 11417885 TI - A history of mammalian embryological research. AB - Although Reinier DE GRAAF (1641-1673) can be considered the founder of modern reproductive biology, scientific knowledge of mammalian development did not progress significantly until the XlXth century. Determining contributions to this progress were the discovery of the ovum by Karl von BAER (1792-1876), his meticulous observations of the stages of embryogenesis, and, half a century later, the remarkable descriptions made by Edouard VAN BENEDEN (1845-1910) of egg development in rabbits and bats. Yet mammalian embryology remained a purely descriptive discipline until the second half of the XXth century, when a handful of exceptional scientists (notably including John D. BIGGERS, Ralph BRINSTER, Anne McLAREN, and W. WHITTEN) managed to obtain reproducibly the development of mouse eggs in a chemically defined medium and to transfer the eggs to the uterine horns of pseudopregnant females. Around the same time (1959), M.C. CHANG was the first to obtain a mammal (a rabbit) by in vitro fertilisation, thus opening the way to assisted procreation. This was achieved in our species in 1978, by Robert EDWARDS and Patrick STEPTOE. With these feats, mammalian embryology could at last become causal, as A. BRACHET already in 1912 had hoped it would. New concepts soon emerged from the delicate manipulations performed on mouse eggs by scientists such as A. TARKOWSKI, B. MINTZ, J. MULNARD, and R. GARDNER, concepts such as the oustside-inside hypothesis proposed to explain the determination of the ICM and trophectoderm or the clonal theory of cell determination during development. These new ideas were soon to become the focus of intense study. Other investigators, interested in the synthesis and roles of macromolecules, contributed in the late 1960's most of our knowledge on global trends in gene expression during the first stages of development. As for the many unfruitful attempts to obtain artificial parthenogenetic development in mice, these would lead to the discovery of parental genetic imprinting. In the 1950's, Leroy STEVENS and Barry PIERCE made famous a very rare tumour, the teratocarcinoma. This tumour soon became a model for studying mammalian development, adopted by an increasing number of research groups. It became the source of a first generation of pluripotent cells culturable in vitro: embryonal carcinoma (EC) cells. In the 1980's came the next generation: embryonic stem (ES) cells derived from the ICM of blastocysts, whose advent coincided with that of the first transgenic mice. Then came the era of knockout mice and cloning. Scientists now envisage with enthusiasm applications that were unimaginable just a few years ago. Such is the legacy of those few pioneers of the experimental embryology of mammals who, in the late fifties, were striving to make the wish expressed by A. Brachet in 1912 come true at last. PMID- 11417886 TI - Research in the canine block. AB - This article, in honor of Dr. Anne McLaren, describes research done during 1955 1959 in the Canine Block at the Royal Veterinary College, London. During that period, Anne in collaboration with the author demonstrated that cultured mouse preimplantation embryos could develop into normal mice after transfer to surrogate mothers. We also studied in depth the control of variability of experimental animals and reproductive aging. In recalling this period, I reminisce about Anne and the scientific environment in which the research was done. PMID- 11417887 TI - From embryo to ethics: a career in science and social responsibility. An interview with Anne McLarne [by Brigid Hogan]. PMID- 11417888 TI - The McLaren effect--a personal view. PMID- 11417889 TI - Anne McLaren as teacher. PMID- 11417890 TI - Anne McLaren--a tribute from her research students. PMID- 11417891 TI - Of microbes, mice and man. AB - This chapter reviews my 18 years of research in Anne's Unit including studies on temporal and spatial aspects of X-chromosome inactivation and imprinting and the role of methylation in X-inactivation in these processes during female mouse embryo development. To enable molecular studies of embryos, we developed a plethora of single cell assays for specific enzyme activity, gene mutation and methylation, and RNA transcription. While in Anne's Unit, I used these same single cell assays to pioneer the procedures for preimplantation diagnosis of genetic disease, now an established clinical approach to prevention of the birth of children with severe genetic disease. At the Institute of Child Health in London, we continue to develop new highly sensitive molecular procedures- currently for the creation of cDNA libraries from human preimplantation embryos, primordial germ cells and embryonal stem cells. We are using these cDNA preparations to isolate human developmental genes and embryo/cancer genes. One of the more fascinating aspects arising from my time in Anne's Unit is the way in which my research findings challenged a number of accepted dogmas in development concerned with the origin and totipotency of the germ line and the possibility of transgenerational genetic inheritance by epigenetic modification of the germ line. PMID- 11417892 TI - Evidence that the testis determination pathway interacts with a non-dosage compensated, X-linked gene. AB - In a number of mammals, including mouse and man, it has been shown that at equivalent gestational ages, males are developmentally more advanced than females, even before the gonads form. In mice, although some strains of Y chromosome exert a minor accelerating effect in pre-implantation development, it is a post-implantation effect of the difference in X chromosome constitution that is the major cause of the male/female developmental difference. Thus XX females are retarded in their development by about 1.5 h relative to X(M)O females or XY males; however, they are more advanced than X(P)O females by about 4 h. It has been suggested that this early developmental difference between XX and XY embryos may "weight the dice" in favour of ovarian and testicular development, respectively, although expression of Sry will normally overcome any such bias. Here we test this proposal by comparing the relative frequencies of female, hermaphrodite and male development in X(P)O, XX and X(M)O mice that carry an incompletely penetrant Sry transgene. The results show that testicular tissue develops more frequently in XX,Sry transgenics than in either of the two types of XO transgenics. Thus the incidence of testicular development is affected by X dosage rather than by the developmental hierarchy. This implies there is a non dosage compensated gene (or genes) on the X chromosome, which interacts with the testis-determining pathway. Since the pseudoautosomal region (PAR) is known to escape X-inactivation, penetrance of the Sry transgene was also assessed in X(M)Y(*X) mice that have two doses of the PAR but have a single dose of all genes proximal to the distal X marker Amel. These mice showed similar levels of testicular development to X(M)O mice with the transgene; thus the non-dosage compensated X gene maps outside the PAR. PMID- 11417893 TI - The case of the midwife scientist. AB - Genes controlling both testis determining and expression of the male-specific transplantation antigen, HY, are located on the short arm of the mouse Y chromosome, and on the X and Y-linked translocation, Sxr(a). A mutation of Sxr(a) was discovered in a cross between an Sxr carrier male and a T16H/X female. This was designated Sxr(b) and found to affect both the expression of HY and spermatogenesis, but not testis differentiation, thereby disproving Ohno's hypothesis that HY controlled testis determination. Molecular genetic analysis showed the mutation to be caused by fusion of two duplicated genes, Zfy1 and Zfy2, deleting the intervening DNA. This deletion interval, deltaSxr(b), contained a number of genes, each a candidate HY gene. Expression cloning with HY specific T cell clones identified Smcy, Uty and Dby as encoding peptide epitopes of this transplantation antigen. The human homologues SMCY and UTY likewise express HY antigens and these are targets of damaging graft-versus-host (GVH) responses and potentially therapeutic graft-versus-leukaemia (GVL) responses following bone marrow transplantation (BMT). Knowledge of the peptide identity of HY epitopes allows monitoring of immune responses following BMT, using fluorescent tetramers, and also offers the possibility of inducing immunological tolerance. PMID- 11417894 TI - Twenty years of research on primordial germ cells. AB - Just twenty years ago I was preparing a research project centred on establishing methods for the isolation and culture of mouse primordial germ cells (PGCs). The project had been suggested to me by Anne McLaren and was to be developed at the Medical Research Council (MRC) "Mammalian Development Unit" in London under the direction of Anne herself. At that time I was a young postdoctoral researcher at the Institute of Histology and Embryology of the University of Rome "La Sapienza" and did not imagine that my decision to be involved in this project would signal a profound switch in my scientific life. From then on my research would mostly concentrate on primordial germ cell biology. I feel like saying that the modern history of mammalian primordial germ cells began twenty years ago at the MRC Mammalian Development Unit under Anne McLaren's impulse. It is not surprising that among the most active researchers in the last twenty years in studying mammalian primordial germ cells, three, namely Chris Wylie, Peter Donovan and myself, began their studies under Anne McLaren's guidance. Over the years, Anne's suggestions and encouragement were always precious for my studies and her presence marked my most important findings on PGC biology. She often invited me to present the results obtained in my laboratory to workshops and congresses. In the present article some of these results particularly influenced by Anne's teaching and suggestions will be briefly reviewed. PMID- 11417895 TI - Germ cell biology--from generation to generation. AB - Germ cells hold a unique place in the life cycle of animal species in that they are the cells that will carry the genome on to the next generation. In order to do this they must retain their DNA in a state in which it can be used to recapitulate embryonic development. In the normal life cycle, the germ cells are the only cells that retain this ability to recapitulate development, referred to as developmental totipotency. The molecular mechanisms regulating developmental potency are poorly understood. Recently its has been shown that germ cells can be turned into pluripotent stem cells when cultured in specific polypeptide growth factors that affect their survival and proliferation. The ability to manipulate developmental potency in germ cells with growth factors allows the underlying mechanisms to be dissected. Germ cells are also the only cells that undergo the unique reductive division of meiosis. This too is essential for the ability of germ cells to form the gametes that will carry the genome into the next generation. Arguably meiosis is the most important division in the life of a nascent organism. Defects in meiosis can result in embryonic or fetal loss or, if the animal survives, in the birth of an individual with chromosomal abnormalities. Recent advances in our understanding of meiosis have come from knockout mice and studies on genes identified through studies of human infertility. This review will focus on these two key aspects of germ cell biology, developmental potency and meiosis. PMID- 11417896 TI - Epigenetic reprogramming of the genome--from the germ line to the embryo and back again. AB - Mammalian parental genomes are not functionally equivalent, and both a maternal and paternal contribution is required for normal development. The differences between the parental genomes are the result of genomic imprinting--a form of gene regulation that results in monoallelic expression of imprinted genes. Cis regulatory elements at imprinted loci are responsible for directing allele specific epigenetic marks required for correct gene expression. This cis information must be interpreted at various points in development, including in the germline where existing imprints are erased and reset. Imprints must also be maintained during preimplantation development, when the genome undergoes dramatic global epigenetic changes. PMID- 11417897 TI - Differentiation of mouse primordial germ cells into female or male germ cells. AB - Mouse primordial germ cells (PGCs) migrate from the base of the allantois to the genital ridge. They proliferate both during migration and after their arrival, until initiation of the sex-differentiation of fetal gonads. Then, PGCs enter into the prophase of the first meiotic division in the ovary to become oocytes, while those in the testis become mitotically arrested to become prospermatogonia. Growth regulation of mouse PGCs has been studied by culturing them on feeder cells. They show a limited period of proliferation in vitro and go into growth arrest, which is in good correlation with their developmental changes in vivo. However, in the presence of multiple growth signals, PGCs can restart rapid proliferation and transform into pluripotent embryonic germ (EG) cells. Observation of ectopic germ cells and studies of reaggregate cultures suggested that both male and female PGCs show cell-autonomous entry into meiosis and differentiation into oocytes if they were set apart from the male gonadal environments. Recently, we developed a two-dimensional dispersed culture system in which we can examine transition from the mitotic PGCs into the leptotene stage of the first meiotic division. Such entry into meiosis seems to be programmed in PGCs before reaching the genital ridges and unless it is inhibited by putative signals from the testicular somatic cells. PMID- 11417898 TI - The allocation and differentiation of mouse primordial germ cells. AB - Analysis of the lineage potency of epiblast cells of the early-streak stage mouse embryo reveals that the developmental fate of the cells is determined by their position in the germ layer. Epiblast cells that are fated to become neuroectoderm can give rise to primordial germ cells (PGCs) and other types of somatic cells when they were transplanted to the proximal region of the epiblast. On the contrary, proximal epiblast cells transplanted to the distal region of the embryo do not form PGCs. Therefore, the germ line in the mouse is unlikely to be derived from a predetermined progenitor population, but may be specified as a result of tissue interactions that take place in the proximal epiblast of the mouse gastrula. The initial phase of the establishment of the PGC population requires, in addition to BMP activity emanating from the extraembryonic ectoderm, normal Lim1 and Hnf3beta activity in the germ layers. The entire PGC population is derived from a finite number of progenitor cells and there is no further cellular recruitment to the germ line after gastrulation. The XX PGCs undergo X inactivation at the onset of migration from the gut endoderm and re-activate the silenced X-chromosome when they enter the urogenital ridge. Germ cells that are localised ectopically in extragonadal sites do not re-activate the X-chromosome, even when nearly all germ cells in the fetal ovary have restored full activity of both X-chromosomes. XXSxr germ cells can re-activate the X-chromosome in the sex reversed testis, suggesting that the regulation of X-chromosome activity is independent of ovarian morphogenesis. PMID- 11417899 TI - Germ cells, gonads and sex reversal in marsupials. AB - The formation of the testis or ovary is a critical step in development. Alterations in gonadal development during fetal or postnatal life can lead to intersexuality or infertility. Several model systems have been particularly useful in studying gonadal differentiation, the eutherian mammal and amphibia, fish, and birds. However, marsupials provide a unique opportunity to investigate gonadal development and the interactions of genes and hormones in gonadal differentiation and germ cell development in all mammals. On the one hand the genetic mechanisms appear to be identical to those in eutherian mammals, including the testis-determining SRY gene. On the other hand, marsupials retain in part the plasticity of the amphibian gonad to hormonal manipulation. It is possible to induce female to male and also male to female gonadal sex reversal in marsupials by hormonal manipulation, and oestradiol can induce male germ cells to enter meiosis at the time the oogonia do. In addition, in marsupials the development of the scrotum and mammary glands are independent of testicular androgens and instead are controlled by a gene or genes on the X-chromosome. Thus marsupials provide a number of opportunities for manipulating the sexual differentiation of the gonads that are not possible in eutherian mammals and so provide a unique perspective for understanding the common mechanisms controlling sexual development. PMID- 11417900 TI - Association of egg zona pellucida glycoprotein mZP3 with sperm protein sp56 during fertilization in mice. AB - Purified mouse sperm receptor, a zona pellucida glycoprotein called mZP3, binds to plasma membrane overlying acrosome-intact sperm heads (P.M. Wassarman, 1999, Cell 96, 175-183). Some evidence suggests that mZP3 binds to sp56, a protein reported to be associated peripherally with the plasma membrane of acrosome intact sperm heads (J.D. Bleil and P.M. Wassarman, 1990, Proc. Natl. Acad. Sci., USA 87, 7215-7219; A. Cheng et al., 1994, J. Cell Biol. 125, 867-878). Here, we report that membrane vesicles prepared from acrosome-intact sperm contain sp56. When these vesicles are incubated with eggs they inhibit binding of sperm to eggs in vitro (ID50 approximately 50-100 microg protein/ml). On the other hand, a monoclonal antibody directed against sp56 relieves the inhibition of binding of sperm to eggs by membrane vesicles. As expected, incubation of intact sperm with the antibody directed against sp56 inhibits binding of the sperm to eggs. Results of immunoprecipitation of sperm extracts incubated with mZP3, by either a polyclonal antibody directed against mZP3 or a monoclonal antibody directed against sp56, suggest that mZP3 is specifically associated with sp56. Results of laser scanning confocal microscopy of fixed sperm probed with antibodies directed against either sp56 or a approximately 155 kDa acrosomal protein, suggest that the two proteins are present in the acrosome, but with different distributions. Furthermore, confocal images of sperm, fixed after exposure to purified mZP3 and probed with antibodies against mZP3 and sp56, reveal overlap between mZP3 and sp56 at the surface of the sperm head. The possible implications of these results are discussed in the context of mammalian fertilization. PMID- 11417901 TI - Chimaeras and mosaics for dissecting complex mutant phenotypes. AB - Back at the first half of the 1980s, there was no mammalian experimental embryology in Hungary. One of us, AN, took up the challenge of establishing a small group in the field. In the absence of local information, AN and his former colleague, Andras Paldi (AP), used their tourist passport to visit several laboratories in Western Europe and collect information and advice. This is how AN and AP ended up one day sitting in Anne McLaren's office in the MRC Mammalian Development Unit at University College, London. They never forgot her endless enthusiasm and the way she clearly explained the important points of preimplantation embryo manipulation, chimaera making and embryo transfer. As well as the extremely useful suggestions, which were crucial to starting the lab in Hungary, they also took back her deep love for embryo development. They remember her telling them, 'never waste an embryo--there is always another unanswered question it can solve'. Many who have been lucky and experienced Anne's spirit and advice later realized how useful it was to generate 'new' ideas by following the 'not wasting' principle. Our views on chimaeras presented below definitely contain elements which grew out from this principle. PMID- 11417902 TI - Size regulation does not cause the composition of mouse chimaeras to become unbalanced. AB - Mouse chimaeras made by aggregating two 8-cell stage embryos undergo size regulation shortly after implantation. Thus chimaeric pups are approximately normal size at birth despite their origin from two complete embryos. Chimaeras of some strain combinations are genotypically unbalanced such that cells of one strain almost always predominate. For example, the BALB/c inbred strain often makes a low contribution to chimaeras. This genotypic imbalance in the composition could arise by selection against BALB/c cells. Selection may be particularly acute at the time of size regulation. To investigate if the mechanism(s) responsible for size regulation could cause the low contribution of BALB/c cells, we compared the composition of an unbalanced series of chimaeras, produced by aggregating two complete 8-cell stage embryos, with a similar series of chimaeras made by aggregating two half 8-cell stage embryos. In each case the unbalanced strain combination was BALB/c<-->[(C57BL x CBA/Ca)F1 x TGB] and parallel studies were undertaken with a genotypically balanced strain combination. For each chimaera, the composition of the fetus, placenta and extraembryonic membranes were determined at E12.5. When two half embryos were aggregated the BALB/c strain still made a poor contribution to all the tissues of the mid-gestation conceptus. This implies that this strain combination remained unbalanced even when size regulation was absent or minimal. Therefore, size regulation did not play a major role in reducing the contribution of BALB/c cells and producing the phenotypic imbalance in the chimaeras. PMID- 11417903 TI - Mouse singletons and twins developed from isolated diploid blastomeres supported with tetraploid blastomeres. AB - The aim of this study was to obtain mice, hopefully identical multiplets, from single diploid blastomeres isolated at the 4-cell stage, or from pairs of sister blastomeres isolated at the 8-cell stage. To this end isolated blastomeres were aggregated with one or two tetraploid carrier embryos produced by electrofusion of 2-cell embryos. Diploid embryos were albino and homozygous for the "a" allele of glucose-phosphate isomerase (GPI-1a1a) and tetraploid embryos were pigmented and GPI-1b1b. The aggregates were cultured in vitro up to the blastocyst stage. Each quartet (occasionally triplet or doublet) of chimaeric blastocysts was transplanted to the oviduct of a separate pseudopregnant recipient. Altogether 62 blastocysts were transplanted to 17 recipients. Eight full-term foetuses (two singletons and three pairs of twins) were rescued by Caesarian section on day 19, 20 or 21 of pregnancy. Three young (one singleton and twins) were successfully reared by foster mothers and proved to be normal and fertile females. All foetuses and animals were albino. In five individuals only the 1-A form of GPI (characteristic for 2n blastomere) was found. In one adult female traces of the 1 B form of GPI (characteristic for 4n carrier blastomeres) were detected in the heart and the lungs while 4 other organs contained only the 1-A form. These observations strongly suggest that the majority of foetuses/animals produced according to our experimental system are 'pure' diploids rather than 2n/4n chimaeras, and that the described method can be used in future to produce twins, triplets and quadruplets in the mouse. Our study confirms earlier work by Kelly (1975, 1977) that 'quarter' blastomeres of the mouse are still totipotent. PMID- 11417904 TI - Implantation: molecular basis of embryo-uterine dialogue. AB - Implantation is a complex developmental process that involves an intimate "cross talk" between the embryo and uterus. Synchronized development of the embryo to the blastocyst stage and differentiation of the uterus to the receptive state are essential to this process. Successful execution of the events of implantation involves participation of steroid hormones, locally derived growth factors, cytokines, transcription factors and lipid mediators. Using gene-targeted mice and a delayed implantation model, our laboratory has been exploring potential interactions among steroid hormones, growth factors, cytokines and prostaglandins in this process. This review article highlights some of our recent observations on the roles of estrogen, catecholestrogen, the EGF family of growth factors, leukemia inhibitory factor and cyclooxygenase-2 derived prostaglandins and their interactions in embryo-uterine "cross-talk" during implantation. PMID- 11417905 TI - Preimplantation genetic diagnosis and embryo research--human developmental biology in clinical practice. AB - Research on human preimplantation embryos in vitro is controversial. Yet it has been the cornerstone for the development important clinical assisted conception techniques. Preimplantation genetic diagnosis which has developed out of this assisted reproductive technology for the first time provides a realistic alternative to prenatal diagnosis and abortion for couples who are at substantial risk of conceiving a pregnancy affected by a known genetic disorder. It also provides the first real hope of therapy for couples who have suffered repeated miscarriages due to chromosome translocations. However, the ability to test very early embryos in vitro presents new and unusual ethical challenges for clinicians and developmental biologists. PMID- 11417906 TI - Reproductive ageing and the menopause. AB - This brief review describes early work initiated by Anne McLaren and John Biggers, in which they repeated on mice a very early experiment carried out by John Hunter on pigs, to test the effect of unilateral ovariectomy on subsequent breeding performance. This and subsequent experiments led to the conclusion that reproductive ageing in the female mouse was largely due to ageing changes in the uterus. As a result of these changes fewer implanted blastocysts are carried to term in the older females, with the result that the size of litters produced gradually drops and ceases altogether well before the expected time of death, thus leading to a period of reproductive inactivity at the end of life. Other organs undergo ageing changes but it appears to be those in the uterus which limit reproductive performance in the female. The somatic organs concerned in bringing the male gametes into the environment are still able to function effectively almost until the time of death so that males have a very short period of reproductive inactivity at the end of their lives. Due to the prenatal onset of meiosis in the germ cells, female mammals and some, but not all, other vertebrates are born with a finite crop of oocytes in the ovary, which cannot be increased after birth. Nevertheless, with the exception of women, female mammals appear to be able to produce ova well into old age, and have them fertilized. When examined after death the ovaries still contain oocytes so this is not a limiting factor in reproduction in old females. In women the situation is completely different. They also have an extended period of reproductive quiescence in middle and old age, the menopause, but, unlike other female mammals, this is not due to failure of the uterus but is caused by the ovary becoming depleted of oocytes in middle age. The reason women run out of oocytes before the end of life, whereas the other mammals which have been studied do not, is associated with the greatly extended lifespan of humans compared to other mammals of equivalent size. There is a linear relationship between longevity and body weight in mammals, small mammals have much shorter lives than large ones. This is probably associated with the increased production of free radical oxygen necessary to maintain body temperature in smaller animals. Heat is lost through the body surface which becomes relatively less as the animal increases in weight, so the smaller animal has to metabolise and thus produces more free radical oxygen to maintain body temperature. For reasons unknown this seems not to apply to humans. The menopause has thus evolved as a consequence of two adaptations: the prenatal onset of meiosis, common to all mammals and many other vertebrates and the greatly increased longevity of all humans, both male and female. In view of this dual origin it is unlikely to have evolved in response to an adaptive need to have grandmothers to help rear the young, as has been suggested! PMID- 11417907 TI - In situ hybridization to mRNA: from black art to guiding light. AB - In situ hybridization to mRNA in embryo sections or wholemount embryos is one of the most powerful analytical tools available to the molecular developmental biologist. For many workers, this procedure provides the first insights into the function of newly isolated genes, allowing the formulation of hypotheses and setting the course for further research. This paper presents a personal historical perspective of the development of in situ hybridization, looks at the theory and practice of the technique, summarizes the current state of the art, and speculates on possible directions for the future as a tool in functional genomics. PMID- 11417908 TI - Effects of Saiko-ka-ryukotsu-borei-to in patients with hyperlipidemia. AB - We measured and compared levels of platelet-derived microparticles (PMPs), monocyte-derived microparticles (MMPs), CD62P on activated platelets, soluble E selectin (sE-selectin), and anti-oxidized low density lipoprotein (LDL) antibody in hyperlipidemia patients and control subjects. Binding of anti-GPIIb/IIIa and anti-GPIb monoclonal antibodies to platelets was not significantly different between hyperlipidemia patients and controls. However, expression of CD62P on platelets and levels of PMPs were higher for hyperlipidemia patients than in controls, although the difference between groups in CD62P expression was not significant (PMPs: 534 +/- 63 vs. 388 +/- 47, p < 0.05; CD62P: 9.1% +/- 1.45 vs. 7.3% +/- 1.15, N.S.). Although there were no differences in expression of CD36 and CD40 by monocytes between the two groups, levels of MMPs were higher in hyperlipidemia patients than in controls (MMPs: 147 +/- 21 vs. 59 +/- 8, respectively, p < 0.01). Levels of anti-oxidized LDL antibody and sE-selectin were also higher in hyperlipidemia patients. We studied the effects of Saiko-ka ryukotsu-borei-to on levels of these factors in patients with elevated triglyceride levels. After Saiko-ka-ryukotsu-borei-to treatment, levels of CD62P, PMPs, sE-selectin, and anti-oxidized LDL antibody were reduced significantly. Levels of triglycerides, total cholesterol and MMPs also decreased, but the changes were not significant. These findings suggest that Saiko-ka-ryukotsu-borei to prevents the development of vascular complications in hyperlipidemia patients. PMID- 11417909 TI - Diallyl sulfide--a flavour component from garlic (Allium sativum) attenuates lipid peroxidation in mice infected with Trichinella spiralis. AB - We investigated the antioxidant capacity of diallyl sulfide (DAS) in the course of experimental trichinellosis in C3H mice. For this purpose, the mice were orally infected with either Trichinella spiralis larvae (300 larvae/mouse) or treated per os with normal saline (control), and 12 hrs later, they were treated per os with DAS (200 mg/kg b.w) daily for 5 or 20 days. On day 5 and 35 of the post-infection period (5 and 35 dpi), thiobarbituric-acid reactive substances (TBARS) in the small intestinal mucosa and diaphragm samples and the total antioxidant status (TAS) of blood were determined. Trichinella spiralis increased TBARS and decreased TAS in the intestinal phase of invasion. However, in the muscular phase mice, neither TBARS nor TAS was found to be different from those of the control. Diallyl sulfide has been shown to decrease TBARS and the agent did not have any effect(s) on the total antioxidant status of blood in Trichinella-infected mice. The results suggest that diallyl sulfide may be an effective antioxidant candidate and may therefore play a significant role in the defense against lipid peroxidation in trichinellosis. PMID- 11417910 TI - Effects of Panax quinquefolius L. on brainstem neuronal activities: comparison between Wisconsin-cultivated and Illinois-cultivated roots. AB - Brainstem neurons receiving subdiaphragmatic vagal inputs were recorded in an in vitro neonatal rat brainstem-gastric preparation. Aqueous extracts of American ginseng root (Panax quinquefolius L.) from Wisconsin and Illinois were applied to the gastric compartment or the brainstem compartment of the bath chamber to evaluate the peripheral gut or central brain effects of the extracts on brainstem unitary activity. After P. quinquefolius extract application to the gastric or brainstem compartment, a concentration-related inhibition in neuronal discharge frequency in brainstem unitary activity was observed, suggesting that P. quinquefolius plays an important role in regulating the digestive process and modulating the brain function in the rat. In this study, pharmacological effects of Wisconsin-cultivated P. quinquefolius and Illinois-cultivated P. quinquefolius were compared. Our results showed that Illinois-cultivated P. quinquefolius possesses a significantly stronger peripheral gastric as well as central brain modulating effect on brainstem neuronal activity. Data from our high performance liquid chromatography ginsenoside analysis suggest that this increase in inhibitory effects by Illinois-cultivated P. quinquefolius may be due to its different ginsenoside profile. PMID- 11417911 TI - The effect of a peat-based preparation on mitogen-induced proliferation of thymocytes in non-treated and hydrocortisone-suppressed mice. AB - The studies were carried out on Balb/c mice (5-6 weeks of age) treated with a peat-based preparation (PBP), administered i.p. once or four times at 24 h intervals at doses of 0.01; 0.1 or 1 mg/kg. Additionally, hydrocortisone was injected i.p. to selected mice at a single dose of 125 mg/kg. The results show that PBP temporarily enhances the proliferative capability of murine thymocytes stimulated in vitro with concanavalin A (Con A) and phytohemagglutinin (PHA). The effect of PBP depends on the number of subsequent doses, but does not depend on the dose applied. A single PBP administration does not affect the proliferative response of thymocytes to Con A and PHA. A single injection of PBP (doses from 0.01 to 1 mg/kg) does not change the number of thymic cells and weight ratio of this organ. Increased doses of subsequent PBP injections (0.01 and 0.1 mg/kg) do not affect the number of thymocytes, but temporarily increase the weight ratio of the thymus two days after the last injection. Administration of PBP prior to hydrocortisone prevents the suppressive effect of the drug on proliferative response of thymocytes stimulated in vitro with Con A and PHA, at the same time increasing the proliferative response of thymic cells to the two mitogens in relation to the control group (hydrocortisone-free). The effect of a single dose of PBP depends on the dose applied--the weakest preventive effect was observed at a dose of 0.01 mg/kg. An increase in the number of subsequent PBP doses, irrespective of a dose applied, prolongs the protective action of the drug on proliferative activity of thymocytes stimulated in vitro with these mitogenes. Moreover, the results obtained in the studies show that PBP partially prevents the suppressive effect of hydrocortisone, as the number of thymic cells and weight ratio of this organ drastically decreased. PBP accelerates regeneration of the thymus, but this depends on a dose applied and the number of subsequent doses. The result was the strongest and the fastest when PBP was injected four times at a dose of 1 mg/kg. It seems quite likely that the thymic regeneration due to PBP is connected with the effect of this drug on maturation and differentiation of thymic cells. PMID- 11417912 TI - The beneficial effect of OST-6 (OsteoCare), a herbomineral formulation, in experimental osteoporosis. AB - OST-6 (OsteoCare), a herbomineral formulation, was evaluated for its inhibitory effect on the progress of bone loss induced by ovariectomy in rats. Ovariectomized (Ovx) rats were administered with OST-6 at 250 and 500 mg/kg b.wt., orally daily for 90 days. On 91st day, ovariectomized rats showed reduced bone mineral content and increased serum alkaline phosphatase levels, excretion of urinary calcium and pyridinium cross-links levels. Histologically, bone sections revealed narrowed and disappearance of trabeculae and widened medullary spaces. The total numbers of Tartrate-resistant acid phosphatase (TRAP) positive cells were significantly increased both in-vivo and in-vitro methods. OST-6, at a dose of 500 mg/kg, significantly improved bone mineral contents, serum alkaline phosphatase levels, reduced the elevated urinary calcium and pyridinium cross links excretion, number of TRAP positive cells and reversal of the above mentioned histological features. These results indicate the usefulness of OST-6 in the management of osteoporosis in a natural way through herbal resources. PMID- 11417913 TI - Involvement of NMDA receptors in the analgesic properties of psychotridine. AB - We have previously reported that the alkaloid extract of Psychotria colorata (Willd. ex R. & S.) Muell. Arg., had marked dose-dependent, opioid-like activity. Phytochemical analyses of P. colorata flowers and leaves identified several pyrrolidinoindoline alkaloids, including psychotridine. To further investigate the activity and mechanism of action of Psychotria alkaloids, we studied the effects of psychotridine on thermal and chemical models of analgesia. In the tail flick model, psychotridine presents a dose-dependent analgesic effect; the effect is not reversed by prior treatment with naloxone. Psychotridine dose-dependently decreased capsaicin-induced pain. Performance in the rotarod test showed that psychotridine does not induce motor deficits at doses effective in analgesia models. Psychotridine inhibited [3H]MK-801 (dizocilpine) binding to cortex membranes in a dose-dependent manner. Binding is completely abolished at 300 nM. The data rule out opioid activity, and the inhibition of capsaicin-induced pain and of radioligand binding strongly suggest the participation of NMDA receptors in psychotridine-induced analgesia. PMID- 11417914 TI - Anti-oxidation activity of different types of natural Cordyceps sinensis and cultured Cordyceps mycelia. AB - Cordyceps, one of the well-known traditional Chinese medicines, consists of the dried fungus Cordyceps sinensis growing on the larva of the caterpillar. It is commonly used for the replenishment of body health. One of the known pharmacological effects is its anti-oxidation activity. However, there is a great variation of the quality in different sources of Cordyceps. Here, the water extracts of various sources of natural C. sinensis and cultured Cordyceps mycelia were analyzed for their anti-oxidation activity by using three different assay methods such as the xanthine oxidase assay, the induction of hemolysis assay and the lipid peroxidation assay. The results showed that Cordyceps, in general, possesses a strong anti-oxidation activity in all assays tested. However, both natural and cultured Cordyceps showed the lowest inhibition in the lipid peroxidation when compared with the other two assay methods. The cultured Cordyceps mycelia had equally strong anti-oxidation activity as compared to the natural Cordyceps. Besides, the anti-oxidation activities were increased to 10-30 folds in the partially purified polysaccharide fractions from the cultured Cordyceps mycelia, which suggested that the activity could be derived partly from Cordyceps polysaccharides. PMID- 11417915 TI - Evaluation of hepatoprotective activity of legumes. AB - Mung bean, adzuki bean, black bean and rice bean are foods and folk medicines of Taiwan. We evaluated the effects of various water extract concentrations (100, 500 and 1000 mg/kg body wt.) and silymarin (25 mg/kg body wt. on acetaminophen induced liver injury by measuring serum glutamate-oxalate-transaminase (sGOT) and serum glutamate-pyruvate-transaminase (sGPT) activities in rats. The results showed that the sGOT and the sGPT activities, increased by APAP, were decreased significantly (P < 0.05) through treatment with inceasing amounts up to 1000 mg/kg body wt. of the exracts. In particular, the mung bean aqueous extract showed the best hepatoprotective effect on APAP-induced hepatotoxicity. The pathological changes of liver injury caused by APAP improved by the treatment with all of the legume extracts, which were compared to silymarin as a standardized drug. In addition to these results, the extract of mung bean acted as a potential hepatoprotective agent in dietary supply. PMID- 11417916 TI - Hepatoprotective activity of Cassia fistula leaf extract. AB - Hepatoprotective activity of the n-heptane extract of Cassia fistula leaves was investigated by inducing hepatotoxicity with paracetamol in rats. The extract at a dose of 400 mg/kg body wt. exhibited orally, significant protective effect by lowering the serum levels of transaminases (SGOT and SGPT), bilirubin and alkaline phosphatase (ALP). The effects produced were comparable to that of a standard hepatoprotective agent. PMID- 11417917 TI - Anti-inflammatory and spasmolytic activity of extracts from Droserae herba. AB - An ethanolic extract of Drosera madagascariensis inhibited human neutrophil elastase with an IC50 of 9.4 microg/ml. The naphthoquinones present in the extract were not responsible for this effect, but flavonoids like quercetin (IC50 0.8 microg/ml), hyperoside (IC50 0.15 microg/ml) and isoquercitrin (IC50 0.7 microg/ml) contributed to inhibition of the enzyme. In guinea-pig ileum the extract (0.5-1 mg/ml) induced a spasmolytic effect via affecting cholinergic M3 receptors and histamine H1 receptors, respectively. At contractile prostanoid receptors of guinea-pig trachea the Drosera extract was not effective. PMID- 11417918 TI - Evaluation of the biological activity of crude extracts from plants used in Yucatecan traditional medicine. Part II. DNA-interacting activity. AB - Extracts from leaves, stems, and roots of twelve plants used commonly in Yucatecan traditional medicine were evaluated in the DNA-methyl green assay. Twenty one extracts showed DNA-interacting activity, and nine of them, belonging to five plant species, presented a displacement activity of 5% or higher. The highest activity (17.6%) was detected in the leaf extract of Heliotropium angiospermum. PMID- 11417919 TI - In vitro keratinocyte antiproliferant effect of Centella asiatica extract and triterpenoid saponins. AB - Psoriasis is a hyperproliferative skin disorder estimated to be present in 1-3% of most populations. Conventional therapy using corticosteroids, Vitamin D analogs and cytotoxic agents eg psoralens is associated with low success rate and many side effects. Traditional plant remedies may provide leads for new treatments. A rapid-throughput, in vitro bioassay has been utilised to examine plants for inhibitory effects on the growth of SVK-14 keratinocytes. Centella asiatica, a reputed anti-psoriatic herb, has been compared against the psoralen containing seeds of Psoralea corylifolia and the synthetic anti-psoriatic agent dithranol (anthralin). Aqueous extracts of Psoralea corylifolia and Centella asiatica inhibited keratinocyte replication with IC50 values of 18.4 +/- 0.6 microg/ml and 209.9 +/- 9.8 mg/ml respectively prior to treatment with polyvinylpolypyrrolidone (PVPP) and 36.3 +/- 3.3 mg/ml and 238.0 +/- 2.5 mg/ml respectively after PVPP treatment of the extracts. The effect produced by C. asiatica is thus unlikely to be due to phenolic compounds. It may, however, be due to its two constituent triterpenoid glycosides madecassoside and asiaticoside which had IC50 values of 8.6 +/- 0.6 microM respectively. These values were comparable to their concentrations in the crude extract and to the IC50 of dithranol (5.1 +/- 0.4 microM). These results suggest that the potential use of C. asiatica extracts as a topical anti-psoriatic agent is worthy of further investigation. PMID- 11417920 TI - Inverse relation between stimulus intensity and seizure duration: implications for ECT procedure. AB - A retrospective analysis of the effects of electroconvulsive therapy (ECT) was performed for two groups of 11 patients matched according to age (mean age, 52 years), sex, and diagnosis. Group 1 received ECT according to the age-dose protocol; group 2 was treated according to the titration method. A higher dose relative to the seizure threshold appeared to shorten the seizure duration. At the first treatment, the correlation between stimulus intensity and seizure duration was negative. In the titration group, the initial mean charge of 91 mC resulted in a seizure duration of 51 s, whereas in the age-dose group the seizure duration of 31 s was significantly shorter despite a higher mean charge of 312 mC. Seizure duration decreased during the ECT course in the group treated first at low dose (titrated) and then at 2.5 times the initial threshold. High stimulus intensity represented adequate treatment, although it produced short seizures. Thus, seizure duration proved to be an unreliable guideline for effective treatment. Furthermore, focus on seizure duration led to frequent high-dose restimulation in the elderly. The titration method obviates inadequate or excessive charges because the seizure threshold must first be determined. PMID- 11417921 TI - Electroconvulsive therapy for minors: experiences and attitudes of child psychiatrists and psychologists. AB - OBJECTIVE: To estimate knowledge, experience, and attitudes towards the use of electroconvulsive treatment in minors (patients < 18 years of age), among child and adolescent psychiatrists and psychologists. METHOD: 1,600 questionnaires were mailed to a group of child and adolescent psychiatrists and psychologists. RESULT: There were 625 (39%) respondents. 329 (53.8%) of the respondents stated that they possessed minimal knowledge about the use of ECT in children and adolescents. Lack of confidence in providing a second opinion was common and reported by 75%. Compared with those with minimal knowledge, respondents with advanced knowledge reported a higher perception of safety and efficacy. The majority (70%) of the respondents regarded ECT as a treatment of last resort. CONCLUSION: Many child and adolescent psychiatrists and psychologists have very little knowledge, training, or experience in this treatment. They seem to be ill equipped to appropriately consider or advise patients and families about ECT. Clinical and research implications of these findings are discussed. PMID- 11417922 TI - Characteristics of responders and nonresponders to brief-pulse right unilateral ECT in a controlled clinical trial. AB - In a double-blind, randomized controlled study of electroconvulsive therapy (ECT) in patients with major depression, 7 of the 17 patients allocated to the right unilateral group failed to respond to treatment. The nonresponders were subsequently openly treated with bitemporal treatment, which produced an acceptable outcome in these cases of right unilateral treatment failure. This paper describes the clinical outcome, electrophysiological characteristics (impedence, estimated seizure threshold, and change in threshold), and the degree to which stimuli exceeded threshold in the responder and nonresponder groups. Responders had lower seizure thresholds and longer seizures than nonresponders. In comparison with nonresponders, responders showed trends toward greater impedance and treatment at a somewhat greater degree above threshold during the first few treatments. Threshold change with treatment was found not to be related to clinical outcome. Early identification of patients likely to respond to low dose right unilateral ECT, together with the avoidance of benzodiazepine prescription during ECT, may permit many patients to receive low-dose right unilateral ECT successfully and with a minimum of cognitive impairment. PMID- 11417923 TI - Coadministration of melatonin and fluoxetine does not improve the 3-month outcome following ECT. AB - At least 50% of patients with manic depressive disorder (MDD) treated successfully with electroconvulsive therapy (ECT) will experience a relapse within the first year of follow-up. Sleep disturbances are very common in MDD and may constitute forerunners of relapse. In this study we tested the hypothesis that melatonin, a sleep-promoting hormone, would decrease the 3-month relapse rate after successful ECT. We included in the study patients with MDD successfully treated with ECT (post-ECT Hamilton Rating Scale for Depression [HRSD] < or = 10). Patients were blindly randomized to two groups, one receiving fluoxetine + placebo and one receiving fluoxetine + melatonin. Assessments (HRSD, Brief Psychiatric Rating Scale, Global Assessment of Function Scale, Global Depression Scale, Pittsburgh Sleep Quality Index, Mini-Mental State Exam, and pill count) were performed for 12 weeks after ECT. Ten of the 35 patients (28.5%) relapsed during the follow-up period. Relapse rates were similar in both groups of patients. Sleep reports were not improved by melatonin. Patients who achieved a higher functional state post-ECT relapsed less often. We conclude that the addition of melatonin to on-going fluoxetine treatment did not have a beneficial effect either on the 3-month outcome post-ECT or on the sleep reports of these patients. PMID- 11417924 TI - Subtypes of memory dysfunction associated with ECT: characteristics and neurobiological bases. AB - Electroconvulsive therapy (ECT) is an effective treatment for a variety of psychiatric syndromes. However, one of its adverse secondary effects is neurocognitive dysfunction. The aim of this paper is to review different subtypes of memory dysfunction associated with ECT from a neuropsychological perspective. Declarative memory is clearly impaired after ECT. Immediate memory, however, is broadly preserved. Few studies have addressed procedural and incidental memory. Selective memory is impaired, probably due to the disruption of specific brain regions. Some of the possible neurobiological bases of ECT memory dysfunction are discussed in this paper. Synaptic plasticity, the cerebral neurotransmission system, and cerebral metabolism are examined in relation to the dysfunction and subsequent recovery of each memory subtype. PMID- 11417925 TI - Relation between frontal 3-7 Hz MEG activity and the efficacy of ECT in major depression. AB - The efficacy of electroconvulsive therapy (ECT) in major depression has been linked to the accentuation of postconvulsive prefrontal electroencephalography slow-wave activity. We investigated the change in slow-wave activity (0.5-7 Hz) using whole-scalp magnetoencephalographic (MEG) recordings. The 3-7 Hz (theta) activity increased in the right frontal and occipital regions during the course of treatment. After four treatments, the increase of the theta activity in the left frontal cortex correlated with the efficacy of the ECT treatment. Moreover, the change of the ratio of left and right frontal theta activity to occipital theta activity had a positive correlation with the therapeutic effect. These findings suggest that an efficient ECT treatment increases MEG theta activity in the frontal cortex. PMID- 11417926 TI - A unilateral, prolonged, nonconvulsive seizure in a patient treated with bilateral ECT. AB - We report on a case of a 45-year-old man in an episode of major depression with psychotic features treated with bilateral electroconvulsive therapy (ECT). At the eighth treatment, he manifested unilateral, prolonged, nonconvulsive seizure activity on the left side, which lasted 351 seconds longer than seizure activity on the right, and was terminated with intravenous diazepam. This is the first report of a unilateral prolonged seizure. Its occurrence following bilateral ECT was particularly noteworthy. This case also highlights the importance of two channel EEG recording during ECT. Without two recording channels we doubt that this event would have been detected, perhaps resulting in nonconvulsive status epilepticus. PMID- 11417927 TI - ECT and automatic internal cardioverter-defibrillator. AB - Electroconvulsive therapy (ECT) is a relatively safe and effective treatment even in depressed patients with preexisting cardiovascular diseases. However, little is known about its use in patients with automatic internal cardioverter defibrillator (AICD). This case report describes a successful administration of ECT in a treatment-refractory depressed patient with an AICD, where the AICD device was deactivated during each ECT treatments and reactivated immediately upon completion of each treatment. The presence of an AICD should not be a contraindication to receiving ECT. PMID- 11417928 TI - ECT and mental retardation: a review and case reports. AB - Electroconvulsive therapy (ECT) in patients with mental retardation has received limited study and is a subject of controversy. Specific difficulties in using ECT for this patient population include diagnostic dilemmas, difficulties with measuring outcome and monitoring side effects, and problems with professional attitudes. We report our experience with two cases in which ECT was applied to treat severe psychotic and catatonic symptoms. In case 1, a 22-year-old male patient with a history of moderate mental retardation, bipolar disorder, and neuroleptic malignant syndrome was admitted to manage his disruptive behavior and psychotic symptoms. The patient responded well to six bilateral ECTs with diminution of his psychotic symptoms and behavioral disturbances. In Case 2, a 39 year-old female patient with a history of mental retardation, schizoaffective disorder, and catatonic symptoms successfully responded to 11 bilateral ECTs. We conclude that ECT can be used safely and effectively in patients with mental retardation and severe or refractory psychotic symptoms. PMID- 11417930 TI - Computer and the practice of ECT: using the Internet. PMID- 11417929 TI - Inclusion of patients who cannot respond to ECT blocks observation of EEG findings. PMID- 11417931 TI - A failure to find large visible effects of ECT-induced seizures on SPECT scans. PMID- 11417932 TI - Stimulus dosing in electroconvulsive therapy and the threshold multiple method. PMID- 11417933 TI - Is electroconvulsive therapy effective for the depressed patient with comorbid borderline personality disorder? AB - Among the more common current indications for electroconvulsive therapy (ECT) is treatment-resistant depression. Treatment resistance is correlated with a number of factors, including the presence of comorbid personality disorders, such as borderline personality disorder (BPD). A detailed review of the literature was undertaken and very few reports or studies have dealt specifically with ECT in borderline patients. Thirteen original reports on ECT outcome in personality disordered patients were identified. Depressed patients with a personality disorder, particularly BPD, may have a poorer outcome on some measures. However, the available data suggests that depression in these patients can be effectively treated with ECT. The depressed, borderline patient appears to have two distinct disorders, one which is responsive to ECT and the other which is not. Unfortunately, the literature is limited by lack of rigorous randomized treatment studies, lack of long-term follow-up, and other methodological weaknesses. Clinical guidelines are suggested. PMID- 11417934 TI - Medical student attitudes and knowledge about ECT. AB - We report the results of a survey of second-year medical students concerning attitudes and basic knowledge of electroconvulsive therapy (ECT). It appeared that there were significant negative biases against ECT in a portion of the group. Forty percent of the students who participated felt that psychiatrists often misused ECT, while 31% actually thought ECT was used to punish violent or uncooperative patients. Few students knew the typical frequency or duration of treatment or even that it was done under general anesthesia. It was interesting that the group describing themselves as highly knowledgeable about psychiatric illness had a greater bias against ECT. Students in the negative group did not differ in the sources of their information about ECT. The most common sources of this information about ECT were movies or college classes. The results document the need for appropriate coverage of ECT in medical school curriculum. PMID- 11417935 TI - Gender differences in cocaine craving among non-treatment-seeking individuals with cocaine dependence. AB - The purpose of this pilot study was to evaluate potential gender differences in cocaine craving among non-treatment seekers with cocaine dependence. We examined 10 female and 11 male individuals matched by demographic characteristics and severity of drug use; we used a multidimensional questionnaire that assesses various aspects of craving: (a) current intensity, (b) projected intensity, (c) resistance to use cocaine, (d) responsiveness to drug-related conditioned stimuli, and (e) imagined likelihood of use if in a setting with access to drugs. Other instruments utilized were the Hamilton Rating Scale for Depression and Addiction Severity Index. Female subjects had higher total craving scores (p < .05), with post hoc tests showing more present desire to use cocaine and responsivity to drug-conditioned stimuli, along with lower scores on the desire not to use cocaine. In exploratory analyses, we found greater depressive symptomatology (p = .02) and severity of family/social problems (p = .02) in females than their males counterparts. These results suggest that gender may influence different aspects of cocaine craving. As estrogen is purported to modulate craving-related dopaminergic systems, further studies will be needed to confirm these observed gender differences and to investigate their possible mechanisms, particularly estrogen-dopamine interactions and their effect on craving and mood. PMID- 11417936 TI - Mother-infant interaction at 12 months in prenatally cocaine-exposed children. AB - This study examined mother-infant interactions of 12-month-old African-American prenatally cocaine-exposed infants and their mothers. Videotaped observations were made during a free-play dyadic interaction, a brief separation, and a reunion period. Videotapes were coded for maternal and child behaviors during each phase of the procedure. Although there were few differences in interactive behaviors between prenatally cocaine-exposed and nonexposed children and their mothers, children who were prenatally exposed to cocaine ignored their mother's departure (odds ratio [OR] = 3.0, p < .05) during separation significantly more often than nonexposed subjects. In addition, mothers who abused cocaine engaged in significantly more verbal behavior (F(2,104) = 7.00, p < .001) with their children than mothers of nonexposed children. These findings indicate that women who used cocaine during pregnancy may not differ from nonusers in their interactions with their 12-month-old infants. PMID- 11417937 TI - Diversity in relapse prevention needs: gender and race comparisons among substance abuse treatment patients. AB - Attempts to address high relapse rates following substance abuse treatment have focused on identifying relapse prevention needs and development of subsequent relapse prevention programs. Few studies have examined whether women and African Americans have unique relapse prevention needs. Research in this area could provide an initial basis for the development of alternative relapse prevention approaches that could be more appropriate for this pop ulation. This study examined gender and race differences in psychosocial concerns among patients recruited from substance abuse treatment as potential indicators of relapse prevention needs. Participants (N = 331) completed several questionnaires during their first month of substance abuse treatment. Assessment packets included measures of coping, self-efficacy, resource needs, cravings, social influences, exposure, and leisure activities. Analyses focused on gender and race differences in these variables before and after controlling for background characteristics (i.e., age, marital status, income, polysubstance use, treatment type, and problem severity). Gender differences found were that men reported poorer coping skills and more negative social influences and exposure to substances than women; these differences remained significant when controlling for background characteristics. Significant race differences were found on all scales except negative social influences. After controlling for background characteristics, African-Americans reported significantly greater coping skills and self-efficacy than did Caucasians; however, African-Americans also reported greater resource needs in comparison to Caucasians. Results highlight the diversity in psychosocial issues among substance abusers in treatment, particularly between Caucasians and African-Americans. Implications for developing alternative relapse prevention approaches to address this diversity are discussed. PMID- 11417938 TI - The first week after drug treatment: the influence of treatment on drug use among women offenders. AB - Over the last decade, there has been a dramatic rise in the number of women arrested for drug offenses, and many have serious drug abuse problems. Increasingly, these women have been mandated to drug treatment, often in community-based settings. This article examines the impact of the treatment programs on the short-term posttreatment drug use of women offenders (N = 165) leaving two community-based treatment programs in Portland, Oregon. Our analyses indicate that women who abstained from drug use during the first week after treatment were more likely than those who used drugs during this time to have remained in treatment longer, received a plan to make a successful transition out of treatment, avoided associations with other drug users after leaving treatment, and obtained encouragement from individuals and groups in support of abstinence. PMID- 11417939 TI - Rural-urban differences in substance use and treatment utilization among prisoners. AB - Surveys of incarcerated offenders and arrestees consistently report high rates of both alcohol and drug use in this population. This drug-crime connection has highlighted the need to learn more not only about drug treatment effectiveness, but also about drug treatment utilization. While studies have begun to examine drug treatment utilization, most of these studies have been based on urban substance abusers. Little is known about the extent to which urban and rural substance abusers may be different in terms of treatment utilization. This study, therefore, examines differences between urban and rural drug use patterns and treatment utilization among chronic drug abusers to determine whether, and in what ways, rurality may affect substance abuse and treatment seeking. The study examines these issues in a group of chronic drug users who were incarcerated at the time of the study. Findings show significant differences in drug use and treatment utilization of urban and rural offenders. Chronic drug abusers from rural and very rural areas have significantly higher rates of lifetime drug use, as well as higher rates of drug use in the 30 days prior to their current incarceration than chronic drug abusers from urban areas. Nonetheless, being from a very rural area decreased the likelihood of having ever been in treatment after controlling for the number of years using and race. While problem recognition appears to explain much of the effect of very rural residence on treatment utilization for alcohol abuse, the effects of being from a very rural area on seeking treatment for drug abuse remain statistically significant even after controlling for several other variables. The findings point to the importance of providing culturally appropriate education to very rural communities on the benefits of substance abuse treatment and of providing substance abuse treatment within the criminal justice system. PMID- 11417940 TI - Self-reported high-risk locations of drug use among drug offenders. AB - The present study provides a detailed, multiple-choice, self-report analysis of home, work, and other public locations where drug offenders report using drugs. In addition, these settings were examined as a function of gender, ethnicity, type of drug used, and drug abuse/dependence status. The participants for the present study were 462 individuals attending drug diversion programs in southern California. The single most frequently reported location of use was the subjects' living room with a small group of friends. However, heavier users used different drugs across a greater variety of locations. Not surprisingly, drugs were used least at work (though a surprising 47% had used at work). Popular situations of drug use among drug offenders are similar to that of high-risk youth. PMID- 11417941 TI - Environmental predictors of heavy episodic drinking. AB - This study examined the relationship between environmental characteristics and heavy episodic drinking by examining college students' last drinking events. For this study, 409 undergraduate students attending a large public university were randomly selected. Students were asked to report on their last drinking event within the 30-day period prior to being surveyed. There were 274 drinking events analyzed. Bivariate analyses and multiple logistic regression analysis were used to identify contextual factors predictive of heavy episodic drinking in specific drinking events. Drinking with friends, drinking beer and hard liquor, and having many people intoxicated at an event were predictive of heavy episodic drinking events. Dating events were protective of heavy episodic drinking. Implications for future research and prevention are discussed. PMID- 11417942 TI - Federal prison residential drug treatment reduces substance use and arrests after release. AB - OBJECTIVE: The effectiveness of federal prison-based residential drug and alcohol treatment programs was evaluated using event history procedures that addressed the problem of selection bias and included a wide range of control variables. METHODS: The sample comprised 760 treatment subjects and 809 comparison subjects. Treatment subjects were from 20 different prisons of medium, low, and minimum security levels. Comparison subjects were drawn from over 30 prisons. RESULTS: The results indicated that individuals who entered and completed in-prison residential treatment were less likely to experience the critical postrelease outcomes of new arrests and substance use during the first 6 months following release. CONCLUSIONS: Without controlling for selection bias, the effects of treatment would most likely have been attenuated. The results have greater generalizability than other studies of prison-based treatment. This study occurred within a multisite context of 20 programs serving both male and female inmates and operating within different security levels and different geographic regions. PMID- 11417943 TI - A benzodiazepine mood effect scale: reliability and validity determined for alcohol-dependent subjects and adults with a parental history of alcoholism. AB - The Tufts Addiction Research Center Inventory--Morphine Benzedrine Group (ARCI MBG) scale was designed to measure benzodiazepine-induced mood elevation. The reliability and validity of the Tufts ARCI-MBG scale were determined in 64 subjects with a history of alcoholism (HA), a positive history of parental alcoholism, defined as one or both parents meeting DSM-III-R criteria for alcohol dependence (PHP), and matched control subjects. Significant correlations were found for within-day Tufts ARCI-MBG scale scores for all groups and for between day scores for PHP and matched control subjects. Interitem reliability was significant for pooled baseline scores. For HA subjects, correlations between mean Tufts ARCI-MBG scale and Drug Liking scores that were obtained after either alprazolam or diazepam administration were significant. These results suggest that the Tufts ARCI-MBG scale is a reliable test that is a valid measure of benzodiazepine-induced mood elevation. PMID- 11417944 TI - Parental alcoholism as a risk factor for DSM-IV-defined alcohol abuse and dependence in American women: the protective benefits of dyadic cohesion in marital communication. AB - Important trends in research over the past decade indicate that women are as greatly affected by familial alcoholism as are men. Although it is increasingly recognized that the adverse drinking outcomes predicted for adult children of alcoholics (COAs) are not inevitable, and only a small percentage develop alcohol dependence or grow up to be alcoholic, relatively little knowledge exists regarding moderating factors that reduce their vulnerability. This study identifies a multiple mediator latent structural model of the intergenerational transmission of risk for DSM-IV-assessed alcohol abuse and dependence among women COAs in adulthood. The effects of both parental alcoholism and family environment are estimated at three time points spanning 10 years across 5-year intervals (1984, 1989, and 1994) using data from a subsample of 4,449 women in the National Longitudinal Survey of Youth (NLSY). Dyadic cohesion in marital communication (greater marital cohesion, harmony, and less verbal disagreement, discord, and conflict) is a proposed moderating factor that may operate in adulthood to lower the risk of female COAs developing alcohol abuse and dependence. Maximum likelihood standardized estimates of the effects of alcohol mediators measured over time indicate that direct parental effects for adverse outcomes decline when COAs are in their late 20s and early 30s. Indirect parental effects through environmental influences dramatically increase the risk of abuse and dependence among COAs at this time if they have one or more alcoholic siblings, especially an alcoholic sister. Dyadic cohesion and positive interpersonal communication patterns were found to moderate effectively the relationship that existed among parental alcoholism, environmental influences, and adverse alcohol consequences. COAs with satisfactory marital communication also evidenced higher levels of intimacy with their partners, perceived the division of housework to be fairer, shared more responsibilities and burdens of the household, and had less conflict over critical domestic issues than other women COAs. The protective benefits of a good marriage against the risks of alcoholism remained when applied to younger and older subjects, across diverse backgrounds, and after adjusting for other factors such as employment status. PMID- 11417945 TI - Evaluation of a social norms marketing campaign to reduce high-risk drinking at The University of Mississippi. AB - A social marketing campaign to change perceptions of peer drinking norms was conducted by the National Golden Key Honor Society at the University of Mississippi during the 1995-1996 school year. To assess the campaign's impact on perceptions of student drinking norms and alcohol consumption, Golden Key's national office administered a survey three times during the school year to all students enrolled in a random sample of required freshmen English courses. Regression analyses suggest that exposure to the marketing campaign may be associated with lower (and more accurate) estimates of student drinking norms. While offering promising results, this study was limited due to shortcomings in the research design. Future evaluations of social norms marketing campaigns should adhere to basic evaluation principles, such as using comparison groups, collecting contextual data, using a valid and reliable survey instrument, and ensuring proper survey administration techniques. PMID- 11417946 TI - Electroretinogram and cue-elicited craving in withdrawn cocaine-dependent patients: a replication. AB - BACKGROUND: We previously reported that cocaine-dependent patients with a reduced blue cone b wave electroretinogram (ERG) responses had significantly more cue elicited craving. METHODS: A new series of 21 recently withdrawn cocaine dependent patients completed a craving questionnaire at baseline and following cue exposure; an ERG was also performed. RESULTS: Cocaine-dependent patients with a blunted ERG blue cone response (<0.5 microV) showed greater increases in craving following cue exposure. When subjects were included from our preliminary study (N = 14), these differences became highly significant. CONCLUSIONS: Patients with a reduced ERG response may represent a subgroup more vulnerable to cocaine craving and future relapse. PMID- 11417947 TI - Effects of Opuntia megacantha leaves extract on renal electrolyte and fluid handling in streptozotocin (STZ)-diabetic rats. AB - We previously demonstrated that Opuntia megacantha leaves' extracts can reduce blood glucose levels in diabetes mellitus. For O. megacantha leaves' extracts to have potential in the management of diabetes mellitus, it is necessary to establish its detailed effects on renal function since diabetes is associated with renal fluid and electrolyte disturbances. Therefore, the current study was designed to investigate the influence of the extracts on renal function in male diabetic Sprague-Dawley rats. Rats were made diabetic by an i.p. injection of streptozotocin (STZ, 60 mg/kg in citrate buffer). Vehicle injected animals acted as controls. Separate groups of nondiabetic and diabetic rats were orally administered O. megacantha leaves extracts (20 mg/100 g bw) or normal saline (0.1 ml x 100 g(-1) bw) daily for 5 weeks. Urine volume and total urinary outputs of Na+ and K+ were determined from 24 h samples. O. megacantha leaves' extracts significantly (p < 0.01) increased urinary Na+ output in diabetic and nondiabetic rats resulting in significantly (p < 0.01) low plasma concentration by comparison with untreated animals. Treatment with the extract significantly increased FE(Na+) and GFR in all groups. The urinary K+ outputs in nondiabetic was slightly lowered, but did not reach statistically significance. O. megacantha leaves' extracts did not alter plasma aldosterone and AVP concentrations in diabetic and nondiabetic rats in nondiabetic animals. It is concluded that O. megacantha leaves extracts modulate renal water and sodium handling. The mechanisms are not clear. PMID- 11417948 TI - Renal and metabolic effects of caffeine in obese (fa/fa(cp)), diabetic, hypertensive ZSF1 rats. AB - In Western society, the triad of hypertension, metabolic syndrome and obesity (which caries a high risk for renal disease) is increasing, as is the intake of caffeine. However, no information is available regarding the metabolic or renal consequences of caffeine consumption in this complex disease entity. The purpose of this study was to investigate the effects of chronic caffeine consumption on renal function and metabolic status in obese ZSF1 rats, an animal model of obesity, hypertension and the metabolic syndrome. Fifteen, 18-week-old male, obese ZSF1 rats were randomized to drink tap water (Cont, n = 8) or 0.1% solution of caffeine (Caff, n = 7) for 8 weeks. Metabolic and renal function measurements were performed at baseline and after 4 and 8 weeks of treatment. Caffeine treatment significantly (p < 0.05) reduced body weight, food, and fluid consumption and improved insulin sensitivity (fasting insulin 129.6+/-8.1 vs 97.5+/-3.6 microIU/mL; fed insulin 146.3+/-8.5 vs 110.6+/-3.4 microIU/mL; fasting glucose 138.7+/-13.4 vs 145+/-8.0 mg/dL; fed glucose 373+/-19.4 vs 283.3+/-19.6 mg/dL, Cont vs Caff, respectively). After 8 weeks of caffeine treatment, animals were less glycosuric as compared with control group. Area under glucose curves (AUC-glucose) in oral glucose tolerance test did not differ between the two groups (AUC- glucose: 592.5+/-42.7 vs 589.5+/-20.5 mg/dL x h, Cont vs Caff), whereas caffeine treatment significantly decreased AUC of insulin (AUC-insulin: 257.77+/-12.9 vs 198.0+/-5.9 microIU/mL x h, Cont vs. Caff, p<0.05). No differences were found with regard to plasma triglycerides and glycerol levels; however, caffeine significantly increased cholesterol levels after 4 and 8 weeks (2F-Anova, p<0.001). Moreover, caffeine significantly decreased creatinine clearance after 4 and 8 weeks (CrCl, Cont: 3.5+/-0.4, Caff: 2.2+/-0.2 L/kg/day, p<0.05) and increased protein/CrCl ratio (Cont: 323+/-30, Caff: 527+/-33 mg/L/day). Caffeine treatment for 8 weeks tended to increase plasma norepinephrine levels (p<0.06), but the two groups did not differ with regard to plasma renin activity, blood pressure, renal blood flow or and renal vascular resistance. The study indicates that caffeine improves insulin sensitivity but increases plasma cholesterol levels and impairs renal function in obesity with the metabolic syndrome and hypertension. Our results imply that the health consequences of chronic caffeine consumption may depend heavily on underlying pathophysiology process. PMID- 11417949 TI - Place and role of new membranes in multiple organ failure: an advance or rumors. PMID- 11417950 TI - Dialytic treatment of rhabdomyolysis-induced acute renal failure: our experience. AB - Acute renal failure is the most common complication of rhabdomyolysis, with an 8 20% reported incidence. In particular, rhabdomyolysis associated with acute renal failure is frequently observed in critically ill patients, with a 6-16% reported incidence in Intensive Care Units. Dialytic treatment is necessary to correct hydroelectrolytic imbalance and renal function alterations and it may be a pathogenetic therapy by myoglobin removal. In the present study we evaluated our experience on patients suffering from rhabdomyolysis and acute renal failure subjected to dialytic treatment. We retrospectively studied 28 patients, 17 admitted in our Intensive Care Unit (ICU-patients) and treated by continuous renal replacement therapy (particularly by continuous venovenous hemofiltration, continuous venovenous hemodialysis and continuous venovenous hemodiafiltration) and 11 admitted in our Nephrology Department (NICU-patients) and treated by high efficiency daily hemodialysis. We excluded one ICU-patient from the study because she was affected with lung end-stage neoplasia and it would have been difficult to evaluate the effects of the dialytic treatment on RML biochemical index and on her final outcome. ICU-patients were older, with a mean age of 64 +/- 10 yrs, and were suffering from MODS and typical elderly diseases, such as cardiac and respiratory chronic failure, except from 3 patients with acute liver failure resulting from poisoning, who were relatively younger. In NICU-patients, instead, the mean age was 36 +/- 16 yrs and the causes of RML were narcotic drugs abuse, repetitive seizures and vigorous exercise, more frequently observed in young people. In three relatively older NICU-patients RML was due to lipid lowering drugs assumption. Before starting the dialytic treatment, in ICU-patients CPK plasma level was 2615 +/- 3586, while K+ was 5.10 +/- 1.08 and sCr was 5.69 +/- 4.06 In NICU-patients, on the other hand, CPK was 14273 +/- 9266, while K+ was 5.75 +/- 0.92 and sCr was 5.9 +/- 0.4. ICU-patients mortality rate was 50% (8/16 patients) in spite of the good recovery of renal function and the biochemical RML indexes improvement. In NICU-patients, instead, only one patient died for septic complications (he was a heroin-addict and suffered from overdose syndrome). Early dialytic treatment of RML allows not only to avoid life-threatening complications (first of all the acute renal failure) but moreover it's a pathogenetic treatment because it removes great amount of myoglobin from the plasma. Beside this, continuous renal replacement therapy allows a successful management of critically ill patients with severe hemodynamic conditions. Nevertheless, the final outcome may be very different between ICU- and NICU-patients, with a higher mortality rate in ICU-patients, suffering from MODS. PMID- 11417951 TI - Clinical improvement by increased frequency of on-line hemodialfiltration. AB - BACKGROUND/AIMS: In spite of the better efficiency of on-line hemodiafiltration (HDF) compared with conventional hemodialysis (HD), it is relatively expensive. The aim of this study was to assess the advantages in the biochemical, hemodynamic and clinical effects in uremic patients treated with on-line HDF and with different frequencies of combination high-flux HD. METHODS: One hundred eleven patients were divided into four groups receiving different frequencies of on-line HDF (thrice, twice, once per week) and high-flux HD. RESULTS: Hemodynamic parameters including maximum drop of systolic blood pressure, episodes of symptomatic hypotension and mean saline infusion volumes during dialysis were reduced when frequencies of on-line HDF were increased. Significant improvements in urea kinetic were observed when frequencies of on-line HDF were increased. On line HDF significantly reduced the amount of erythropoietin needed and improved intra- and inter-dialysis symptoms, physical well-being, menstruation and skin pigmentation when frequency of HDF is increased to three time per week. CONCLUSION: On-line HDF offers a better cardiovascular stability and clinical improvement. Thrice weekly on-line HDF offers a significant benefit when compared with lower frequencies of HDF per week and high-flux HD. PMID- 11417952 TI - Organ system failures prediction model in intensive care patients with acute renal failure treated with dialysis. AB - OBJECTIVE: To evaluate the organ system failures hospital mortality predictions in critically ill patients with acute renal failure requiring dialysis. DESIGN: Prospective, cohort study. SETTING: Intensive care units in a tertiary care university hospital in Taiwan. PATIENTS: A total of 112 patients admitted to the intensive care units with acute renal failure who required dialysis from January 1999 through December 1999. INTERVENTIONS: Collection of information necessary to compute the number of failed organs. MEASUREMENTS AND RESULTS: Of the 112 patients studied, 75 were men and 37 were women. The mean age of survivors and non-survivors was 58.59 +/- 19.91 years and 58.76 +/- 19.62 years. The overall mortality rate was 67%. There were no significant differences between survivors and non-survivors in terms of age, gender, or indication for dialysis. The cause of death in the majority of patients was related to organ system failure during the 24 hours immediately preceding the initiation of acute hemodialysis, and carry mortality rates exceeding 83% with the coexistence of four or more failed organs. The area under the organ system failures prediction model receiver operating characteristic curve equaled 0.772 +/- 0.046. CONCLUSION: We conclude that mortality rate for acute renal failure in intensive care unit patients continues to be high. Organ system failures prediction model performed well and simple in its ability to identify patients who die in hospital. Mortality rate increases as number of failed organ increases. PMID- 11417953 TI - Impairment of renal function after cardiopulmonary bypass is not influenced by dopexamine. AB - INTRODUCTION: The objective of this study was to evaluate the effects of dopexamine on renal function in 4 groups of patients either with or without renal dysfunction. Transient renal dysfunction is often not clinically relevant in patients with normal renal function, but it is an important clinical factor in patients with pre-existing renal failure. Dopexamine (DX) is a commonly used catecholamine which probably exerts a selective effect at the splanchnic bed. MATERIAL AND METHODS: 24 patients with normal renal function and 24 patients with impaired renal function (creatinine in serum > or = 1.5 mg/dL) were each randomly allocated to 2 groups. Group 1 (control) without renal dysfunction and group 3 (control/dysfunction) with renal dysfunction were considered as control groups, while the patients in DX and DX/dysfunction groups received 1 microg/kg/min dopexamine until the end of surgery. Kidney function was investigated using standard parameters and by investigating specific proteins and enzymes. RESULTS: All patients showed pathologic excretions of the investigated parameters during cardiopulmonary bypass (CPB) with no differences between the study groups. The distal tubule, the lysosomal regions, Henle's loop and the glomerular tuft were all damaged. Heart rate and cardiac index increased significantly in the DX groups, first until the end of surgery, second until the start of ECC. CONCLUSION: Dopexamine at a dose of 1 microg/kg/min had no influence on renal function and protein excretion and cannot be regarded as a kidney function protecting substance. PMID- 11417954 TI - Evaluation of lipids peroxidation products vs. proinflammatory cytokines in hemodialized patients. AB - BACKGROUND: Patients with end-stage renal failure on chronic hemodialysis often demonstrate accelerated development of atherosclerotic changes and cardiovascular complications. In those patients oxidative stress facilitates the intensity of lipid peroxidation process, expressed as increased products of lipid peroxidation (malonaldehyde and 4-hydroxyalkenals). Simultaneously, structures modified by peroxidation and glycation of autoantigenic character are formed. Additionally, increase in proinflammatory cytokines is found in their patients on hemodialysis. The aim of the study was to find out the relations between the products of lipid peroxidation and proinflammatory cytokines in patients on hemodialysis. METHODS: Plasma concentrations of malonaldehyde and 4-hydroxyalkenals were estimated by spectrophotometric method, glutathione peroxidase as well as concentration of IL 6, its soluble receptor and TNFalpha were measured using ELISA kits. RESULTS: Aldehyde concentrations (malonaldehyde and 4-hydroxyalkenals) were found to be five fold higher in comparison with the controls. Concentrations of proinflammatory cytokines: IL-6, TNFalpha and soluble IL-6 receptor were also higher than in the control group. Patients on hemodialysis showed positive correlation between concentrations of MDA+4HNE and IL-6 and TNFalpha. CONCLUSION: The correlations found between the products of lipid peroxidation and proinflammatory cytokines suggest causative relation between the intensity of peroxidative processes and stimulation of immunological response in hemodialysed patients, which may increase the risk of atherosclerotic changes in those patients. PMID- 11417955 TI - The hormonal and radiological evaluation of adrenal glands, and the determination of the usefulness of low dose ACTH test in patients with renal amyloidosis. AB - Amyloidosis is a multisystem disease which may cause organ loss. Renal involvement is the most common clinical problem in amyloidosis, however involvement of endocrin organs is possible. In this study to assess adrenocortical function and to evaluate the usefulness of low dose ACTH test in patients with renal amyloidosis, we determined cortisol, 17-hydroxyprogesteron (17-OHP) and 11-deoxycortisol (11-DOC) responses to both 1 microg and 250 microg Synacthen. We also determined the size of adrenal glands radiologically by using computerized tomography. Twenty one patients with renal amyloidosis and 16 healthy subjects for hormonal evaluation, and 20 patients with renal amyloidosis and 22 healthy subjects for radiologic evaluation were included in the study. In four patients (19%) peak serum cortisol levels following stimulation with the low dose of Synacthen were less than 20 microg/dL (550 nmol/L). Two of them had also subnormal cortisol response to the 250 microg Synacthen stimulation test. Basal and stimulated levels of 11-DOC were lower than those of control values (p=0.000 and p<0.01 respectively). The mean 11-DOC responses to stimulation with 1 microg Synacthen were also significantly lower than the values obtained after the simulation with 250 microg Synacthen (p<0.01 and p=0.000). Cortisol responses to the stimulation with 250 microg Synacthen were also lower than the control responses (p<0.05). 17-OHP responses were similar to the control values in both tests. In the radiological evaluation the mean maximum width of right adrenal glands and the mean anterior and maximum width of left adrenal glands were significantly greater in the patient group (p<0.01). In conclusion, adrenal involvement and adrenal insufficiency is common in amyloidosis. Low 11-DOC levels in amyloidosis is a new finding and further detailed studies is required to explain its cause. PMID- 11417956 TI - Acute effect of hemodialysis with polyacrylonitrile membrane on nerve conduction velocities. AB - AIM: Non cellulosic membranes, as polyacrylonitrile (AN69) improves middle molecules purification through hemodialysis, and this increased clearance of middle-molecules may have benefical effects on uremic polyneuropathy. We have tried to evaluate this effect comparing nerve conduction velocities before and after hemodialysis with either AN69 or cellulose acetate (CA). PATIENTS AND METHODS: Eight patients in hemodialysis with AN69 for more than three months (4 men and 4 women, mean age 58.4 +/- 12.9 years, mean time in hemodialysis 9.3 +/- 7.5 months). Motor conduction velocities (MCV) and sensory conduction velocities (SCV) were measured predialysis and postdialysis with AN69 and, one week later, predialysis and postdialysis with CA. RESULTS: There were no differences neither in Kt/V values (AN69 1.27 +/- 0.36 vs CA 1.16 +/- 0.26) nor in TAC ones (AN69 40.8 +/- 17.4 vs CA 40.8 +/- 21.3 mg/dL). After hemodialysis with AN69 MCV significantly increased (42.0 +/- 3.0 vs 40.6 +/- 2.6 m/s baseline, p < 0.05). SCV was also enhanced (45.3 +/- 2.2 vs 41.1 +/- 3.4 m/s baseline, p < 0.05). After hemodialysis with CA neither MCV changed (42.7 +/- 1.6 vs 42.0 +/- 2.3 m/s baseline), nor did SCV (45.8 +/- 5.8 vs 44.8 +/- 3.0 m/s baseline). CONCLUSIONS: Hemodialysis with AN69 acutely improved SCV and MCV. This effect was not seen with CA hemodialysis. PMID- 11417957 TI - Continuous renal replacement therapy: our experience in intensive care unit. AB - Continuous Renal Replacement Therapy (CRRT) indication is still discussed. We report our experience on 98 patients affected with Multiple Organ System Failure (MOSF) and renal failure (acute or chronic) requiring dialysis and timely treated by CRRT. Mortality after 5 days of ICU permanence was 60.2%; the remaining 39 patients were discharged within 21 days and received CRRT treatment for 6.36 +/- 5.59 days. APACHE II score was not able to predict the outcome of patients suffering from acute renal failure (ARF). On the contrary, Systemic Inflammatory Response Syndrome (SIRS) incidence was significantly higher in deceased patients compared to recovered patients. In conclusion, it is important to start dialytic treatment immediately when patients affected with MOSF show renal function damage, even if at an initial stage, in order to improve patients' survival. Moreover a multidisciplinary approach is preferable in ICU patients treatment for not underestimating the management of metabolic and infective complications, the nursing care, and nutritional support. PMID- 11417958 TI - Acute renal failure: expect the unexpected. PMID- 11417959 TI - Fatal cerebral hemorrhage and acute renal failure after young Bothrops jararacussu snake bite. AB - An unusual case of a patient developing severe coagulopathy disorder and a clinical picture of cerebral hemorrhage and acute renal failure after young Bothrops jararacussu snake bite is reported. The mechanisms of snake venom induced injury are discussed and similar cases in literature are revised and compared. The use of bothropic-Crotalus antivenom in severe B. jararacussu envenomation is discussed. PMID- 11417960 TI - Secondary hyperparathyroidism and brown tumor in dialyzed patients. AB - Secondary hyperparathyroidism is one of the most common complications of chronic renal failure (CRF). Its pathogenesis is multifactorial and still not completely understood. Pathological mechanism of hypocalcemia, hyperphosphatemia and calcitriol deficiency are basic characteristics of CRF and main reason for morphological changes in parathyroid glands and hyperparathyroidism (HP). We present a case of a female patient born in 1975. At the age of 10, a urinary infection was diagnosed for the first time and treated. Six years later, as nausea and vomiting started, CRF based on bilateral reflux was diagnosed and the patient was included in the hemodialysis treatment. The patient was again examined in 1997, when biochemical parameters, including the level of parathyroid hormone, ultrasonography of the neck, scintigraphy of the skeleton and densitometry revealed secondary HP. Parathyreoidectomy was perfomed in 1998. During the follow up period, a tumefaction on a ramus mandibulae dex. was noticed, which was cytologically diagnosed as osteitis fibrosa, "brown tumor", a rare complication of the secondary HP. Surgery was performed and PHD was granuloma gigantocelulare. Prevention and therapy of secondary HP is a problem that demands early actions to avoid possible complications. PMID- 11417961 TI - Crescentic glomerulonephritis associated with renal cell carcinoma. AB - Various glomerular lesions are described in association with malignancy. We report a rare case of crescentic glomerulonephritis occurring in association with renal cell carcinoma. A granular deposition of immune complexes and complement strongly suggests a tumor associated immune complex glomerulonephritis. PMID- 11417962 TI - Metastatic bone disease and the role of biochemical markers of bone metabolism in benign and malignant diseases. PMID- 11417963 TI - Prognostic and predictive factors in breast cancer. AB - Around 570 000 women develop breast cancer worldwide. In the U.K. it affects 33 000 women and causes 16 000 deaths each year. Treatment of early breast cancer is surgical, comprising breast conserving surgery (followed by radiotherapy) for small unifocal tumours, or mastectomy for larger or multifocal tumours. Survival of patients with breast cancer depends on two different types of prognostic factors: tumour size reflecting how long the tumour has been present, and biological factors (i.e. grade) which represent tumour aggressiveness. In women with a tumour that has adverse features predicting early recurrence (i.e. lymph node positivity, large size, high grade) adjuvant systemic chemo- or hormonal therapy is given to reduce the risk of relapse. Chemotherapy is given to pre menopausal women for oestrogen receptor negative post-menopausal breast cancer, whereas hormone therapy is reserved for oestrogen receptor positive cancer. Since 50% of patients will never relapse, identification of which women are at high risk of recurrence is necessary so as to offer treatment with adjuvant therapy. The use of hormone therapy and chemotherapy has been aided by factors predicting the likelihood of response, e.g. oestrogen receptor status. The value of newer prognostic and predictive markers is addressed. PMID- 11417964 TI - Prognostic and predictive factors in prostate cancer. AB - Prostate cancer is a major public health problem in the Western world, and the second most common male malignancies in the European Union. Detection of the disease is possible at an early stage, using serum prostate specific antigen measurement and prostatic biopsies. To date, however, screening for prostate cancer has not been shown to be of benefit to patients in improving outcome. This is compounded by uncertainties surrounding treatment efficacy, as more men appear to die with prostate cancer than from it. Studies addressing these issues are underway in Europe and the U.S.A. Clinicians are currently unable to advise their patients with any degree of certainty as to the appropriateness of treatment for prostate cancer, because of their inability to differentiate tumours that will progress from those that will remain quiescent. This article reviews the various clinical, pathological and experimental markers available, and their value in providing prognostic information, which may assist clinicians and patients in making management decisions. Further research is still required to understand the biological behaviour of prostate cancer and to assess the value of screening and treatment efficacy in order to advise patients, clinicians and health care systems accordingly. PMID- 11417965 TI - Prognostic factors for skeletal relapse in breast cancer. AB - Bone metastases and the strong interaction between osseous and metastatic cell populations require interdisciplinary thought and actions. If it were possible to interrupt the malignant dialogue between tumour and bone at an early stage, this might not only reduce the amount of bone destruction, but could also reduce the incidence of osseous metastases and remove the source of secondary metastases to other organs. Studies into the preventive effects of bisphosphonates are currently running or are planned. Most of these studies are in breast cancer patients with involvement of the axillary lymph nodes. The prognostic factors of lymph node status, tumour size and grading are better than none, but do not select patients at a high risk of skeletal metastasis. This would be much better done by using immunohistochemical methods to investigate the primary tumour for bone sialoprotein and parathyroid hormone-related protein (PTHrP). However, these methods are complicated, have not been validated in large numbers of patients and are not standardized. Serum tests for bone sialoprotein, PTHrP and collagen fragments are currently still under development and cannot be recommended generally. The clinical importance of tumour cells in the bone marrow has been demonstrated but is still only used at a few centres. PMID- 11417967 TI - Metastatic bone disease: clinical features, pathophysiology and treatment strategies. AB - Metastatic bone disease develops as a result of the many interactions between tumour cells and bone cells. This leads to disruption of normal bone metabolism, with the increased osteoclast activity seen in most, if not all, tumour types providing a rational target for treatment. The clinical course of metastatic bone disease in multiple myeloma, breast and prostate cancers is relatively long, with patients experiencing sequential skeletal complications over a period of several years. These include bone pain, fractures, hypercalcaemia and spinal cord compression, all of which may profoundly impair a patient's quality of life. External beam radiotherapy and systemic endocrine and cytotoxic treatments are the mainstay of treatment in advanced cancers. However, it is now clear that the bisphosphonates provide an additional treatment strategy, which reduces both the symptoms and complications of bone involvement. Ongoing research is aimed at trying to define the optimum route, dose, schedule and type of bisphosphonate in metastatic bone disease and in the prevention and treatment of osteoporosis in cancer patients. In vitro suggestions of direct anticancer activity and some promising clinical data in early breast cancer have resulted in considerable interest in the possible adjuvant use of bisphosphonates to inhibit the development of bone metastases. PMID- 11417968 TI - Assessment of response to systemic therapy focusing on metastatic bone disease. AB - The 25-year-old rules for response evaluation developed by UICC and WHO still remain the cornerstone for response evaluation and documentation of treatment effects from systemic cancer treatment. Often, however, plain radiographs are insensitive to changes in tumour growth and disagree with clinical findings. Other methods of evaluation, such as biochemical markers and newer imaging technology may provide valuable additional information in these cases. PMID- 11417969 TI - Bone markers in the management of metastatic bone disease. AB - Newer markers of bone formation and bone resorption are now available. Alone these new markers do not appear to be useful to diagnose metastatic bone disease. Several markers appear to be of prognostic importance and correlate with the extent of tumour in the skeleton. Serial monitoring of bone markers may be useful to monitor the efficacy of bisphosphonate and/or systemic therapy of both lytic and blastic bone metastases. PMID- 11417970 TI - Markers of bone turnover in prostate cancer. AB - Prostate cancer is the most common malignancy in elderly men and is often associated with bone metastases. Although bone metastases are osteosclerotic, histological and biochemical studies clearly indicate an increase of both bone formation and bone resorption, providing the rational for using bisphosphonate as a palliative treatment in these patients. The recent development of specific and sensitive biochemical markers, reflecting the overall rate of bone formation and bone resorption, has improved the non-invasive assessment of bone turnover abnormalities in patients with prostate cancer. The immunoassays for bone specific alkaline phosphatase and type I collagen propeptides are currently the most sensitive markers to assess bone, formation. The best indices of bone resorption are the immunoassay for the pyridinoline cross-links and the related peptides that can be measured in urine and more recently in serum. A better knowledge of the biochemistry, especially of the age-related post-translational modifications of type I collagen in the abnormal bone matrix, associated with bone metastases from prostate cancer may lead to markers of increased sensitivity. A recent example is the demonstration that the isomerization and racemization of the aspartic acid residue in C-telopeptides of type I collagen is impaired in patients with prostate cancer and bone metastases, a pattern than can be detected with specific conformational antibodies. The most sensitive markers of bone formation and bone resorption are markedly increased in patients with bone metastases compared with patients with cancer but without metastases, the levels correlating with the extent of the bone involvement. However, their sensitivity remains limited, suggesting that the currently available biochemical markers cannot be used as a surrogate for bone scintigraphy in the diagnosis of bone involvement. A few studies have suggested that the measurement of bone markers may be useful in the assessment of response to anti-endocrine therapy, although available data indicate a lower sensitivity than with prostates specific antigen. Additional longitudinal studies are required to assess the potential use of bone markers, especially to identify patients who relapse during the course of the treatment and, more specifically 3 those that result from the progression in bone metastases.Clearly, the established use of bone markers is for monitoring effects of bisphosphonate treatment. Several studies have shown a rapid decrease of bone resorption markers in patients with prostate cancer and bone metastases, the magnitude of the decrease correlating with the efficacy of the treatment in reducing bone pain. Thus, bone markers are likely to become a useful and objective tool to monitor bisphosphonate treatment and individual the therapy scheme. PMID- 11417974 TI - Does c-erbB2/HER2 overexpression predict adjuvant tamoxifen failure in patients with early breast cancer? PMID- 11417975 TI - There may never be a final cure for breast cancer. AB - This paper challenges the perception that the success of the human genome project has brought us to the threshold of a cure for many common cancers. Paradoxically, the message emerging from the genetics of breast cancer may be that the genetic damage associated with malignant transformation, and the mutations predisposing to the disease in our gene pool, may prove impossible to eliminate. PMID- 11417976 TI - Hyperthermia and hypoxia: new developments in anticancer chemotherapy. AB - It has been demonstrated in vitro and in vivo that hyperthermia can enhance the cytotoxicity of some chemotherapeutic agents. The in vivo studies have demonstrated that the thermal advantage is maximized at mild temperatures such as at 40.5--43 degrees C. Thermo-chemotherapy is widely applied in limb perfusion and intraperitoneal chemotherapy. Hypoxia in solid tumours leads to resistance to most anticancer drugs and appears to accelerate malignant progression and increase metastasis. The recent development of new drugs highly toxic to hypoxic cells may bring new strategies in anticancer treatments and move this condition from being a problem to a new tool in cancer control. PMID- 11417977 TI - Genetic cancer syndromes and large-scale gene expression analysis: applications in surgical oncology. AB - The last decade of the 20th century was characterized by an explosion in genetic discoveries. The Human Genome Project and technical advances have made it possible to unravel many genetic abnormalities underlying cancer. Many genes responsible for inherited cancer syndromes have been identified and diagnostic tests are readily available. The clinical implications of these tests are currently under debate. Large-scale gene expression analysis enables simultaneous monitoring of expression of thousands of genes, in vitro and in vivo. The identification of high risk patients and drug responsiveness, can be studied within the framework of complex molecular networks. This article will focus on the possibilities for surgical oncology. PMID- 11417978 TI - The impact of recurrent rectal cancer on quality of life. AB - AIMS: To determine, in patients operated upon for rectal cancer, the impact that tumour recurrence has on quality of life. METHODS: Twenty-five patients, who had curative surgery for rectal cancer from 1992 to 1997, and who subsequently developed tumour recurrence, participated. Each patient was individually matched with two patients who also had rectal cancer surgery during the same period of time but were considered to be free of cancer. The patients were matched for age, gender, type of operation and socioeconomic status. They answered the European Organization for the Research and Treatment of Cancer (EORTC) QLQ-C30 and CR38 quality of life questionnaires, as well as the SF36 II questionnaire, at least 1 year after surgery. RESULTS: Patients with recurrent rectal cancer had significantly lower quality of life scores than cured patients in most dimensions, as measured by the three questionnaires. A significant difference was also seen in the global quality of life score on the QLQ-C30 (median scores 50 vs 75, z = 4.015, P = 0.0001). CONCLUSION: Recurrence of rectal cancer has a profound effect on quality of life, affecting most measurable dimensions. Treatment of recurrent cancer should not only be directed at the physical aspect of the disease, but also at its psychological and social aspects. PMID- 11417979 TI - Relationship between tumour vascularity and circulating cancer cells in patients with colorectal carcinoma. AB - BACKGROUND: Colorectal cancer vascularity correlates with risk of metastasis. Greater tumour vascularity may increase haematogenous dissemination by providing a larger vessel area for tumour cell invasion into the circulation. We assessed whether the prevalence of tumour cells in the circulation of colorectal carcinoma patients (CTC) increased with tumour vascularity. METHODS: Pre-operative blood samples were assessed for circulating tumour cells using RT-PCR for carcinoembryonic antigen (CEA) and cytokeratin 20 (CK20) mRNA. Vessel count and volume were morphometrically assessed from tumour biopsies after vasculature staining. RESULTS: Thirty-three colorectal cancer patients (M:F, 20:13; mean age 66 years, SD 11 years) were studied. One or more blood samples were RT-PCR positive for either CEA or CK20 mRNA or both, in 28 (85%) patients. There were no significant differences in the prevalence of RT-PCR positive patients between high and low tumour vascularity groups, or in tumour vessel counts or volume in RT-PCR positive compared with negative patients. CONCLUSIONS: These results do not support vascularity related variation in access of tumour cells to the circulation as an explanation for the correlation between tumour vasculature and metastasis. Tumour vascularity and metastatic potential may be linked phenotypes rather than cause and effect. PMID- 11417980 TI - Metastatic pattern of CC531 colon carcinoma cells in the abdominal cavity: an experimental model of peritoneal carcinomatosis in rats. AB - BACKGROUND: Peritoneal spread of tumour cells is a major source of morbidity and mortality in patients with colorectal cancer. In order to develop strategies to prevent intraperitoneal dissemination and to treat peritoneal carcinomatosis, the spread of tumour cells in the peritoneal cavity was studied. METHODS: Two million CC531 colon carcinoma cells were administered intraperitoneally in five groups of eight rats. The rats were killed after 1, 2, 4 and 8 hours and 3, 7, 14 and 21 days. After inspection of the abdominal cavity, samples of blood and ascites were taken. Liver, spleen, omentum, mesentery, diaphragm, parathymic lymph nodes and lungs were removed for histology and immunohistochemistry. RESULTS: No abnormalities were seen in the abdominal cavity until day 3. Subsequently the peritoneum and omentum became thickened and after 21 days all rats had haemorrhagic ascites and peritoneal carcinomatosis. The abdominal fluid contained tumour cells at all stages. The number of tumour cells decreased in the first 8 hours, and increased thereafter. At microscopy the peritoneum was completely covered by tumour cells after 3 days. Tumour cells concentrated in the milky spots (MS) of the omentum within 4 hours. The size of the MS increased as a result of an increase in number of tumour cells and macrophages. After 7--21 days the MS were completely replaced by tumour cells and new MS were formed. In the diaphragm tumour cells invaded the lymphatic lacunae after 8 h, and obliterated these after 3--7 days. Also invasion of the muscle fibres was seen after 3 days. Microscopically no tumour cells were found in blood, liver, spleen, parathymic nodes and lung. CONCLUSION: After intraperitoneal administration of CC531 colon carcinoma cells, tumour cells spread throughout the abdominal cavity, and concentrate in the milky spots of the greater omentum, the paracolic gutters, the subhepatic and subphrenic spaces and in the lymphatic lacunae of the diaphragm. PMID- 11417981 TI - Survival and quality of life after palliative surgery for neoplastic gastrointestinal obstruction. AB - OBJECTIVE: The aim was to identify the prognostic factors which relate to the results, in terms of survival and quality of life, of palliative surgery in cancer patients presenting with an occlusion. METHODS: The files of 109 patients with a neoplasm who were operated on for occlusion between 1990 and 2000 have been re-examined. The prognostic factors studied were age, sex, the location of the primary tumour, the extension of the cancer at the time of the operation, and the surgical procedure carried out. The impact on the quality of life was assessed by the resumption of transit and the return home. RESULTS: The median survival was 64 days and the peroperative mortality was 21%. The quality of life of patients has been improved in 65% of cases. The only factors clearly correlating to survival and the success of the operation are the aetiological diagnosis of the occlusion (local recurrence better than carcinomatosis) and the type of procedure it was possible to carry out (resection better than bypass). CONCLUSION: Palliative surgery can, in a certain number of cases, improve the quality of life of patients, but it has not been possible for us to demonstrate prognostic factors which would allow the selection of patients who could benefit the most from such surgery. PMID- 11417982 TI - Chromophore-enhanced 805 nm laser therapy for gastrointestinal neoplasia. AB - BACKGROUND: A new technique of 805 nm semiconductor diode laser-tissue interaction potentiation using indocyanine green (ICG) as an exogenous chromophore has been assessed for use in the palliation of advanced gastrointestinal malignancy. MATERIALS AND METHODS: The tissue effects of ICG potentiated 805 nm laser were assessed both in-vivo using rat stomach and colon, and in vitro using normal and neoplastic human gastric and colonic mucosa. RESULTS: Intravenous ICG did not potentiate the laser-tissue effects of the 805 nm laser in in-vivo studies. Local techniques of ICG application (submucosal injection +/- surface painting of 0.1% ICG solution) enhanced the total depth and diameter of laser induced thermal injury in all in-vitro tissues. Without ICG enhancement thermal tissue injury was minimal in normal gastric and colonic mucosa. CONCLUSION: Local techniques of ICG application enhance laser induced thermal injury in normal and neoplastic gastric and colonic mucosa. This may permit targeting of 805 nm laser-tissue effects with minimal collateral damage during endoscopic laser palliation of advanced gastrointestinal malignancy. PMID- 11417983 TI - Ductal carcinoma in situ of the breast -- among factors predicting for recurrence, distance from the nipple is important. AB - AIMS: To assess local and systemic recurrence rates and factors predicting for recurrence in patients treated for ductal carcinoma of the breast (DCIS). METHODS: Patients with DCIS treated between January 1986 and January 1997 were identified. All pathology specimens were reviewed. DCIS type, lesion size, nuclear grade and margin clearance were assessed. Mammograms were reviewed and mammographic patterns, size, type of lesion and distance from the nipple were measured. Treatments and subsequent outcomes were established by case note review. Factors predicting for recurrence were analysed by both univariant and multivariant analysis. RESULTS: Of the 220 patients, 153 (70%) had breast conserving surgery. Sixty-seven (30%) had a mastectomy. Ninety-seven patients had adjuvant therapy of which 22 had radiotherapy alone, 54 had tamoxifen alone and 21 had radiotherapy and tamoxifen. Following mastectomy, two patients developed axillary recurrences. Following breast-conserving surgery 20 (13%) patients developed local recurrences, of which one developed systemic disease and died from breast cancer. CONCLUSIONS: Mammographic nipple to lesion distance of <40 mm and high/intermediate nuclear grade were the only factors found to increase the likelihood of recurrence. PMID- 11417984 TI - Sentinel lymph-node biopsy compared to axillary lymph-node dissection for axillary staging in breast cancer patients. AB - AIMS: The purpose of this study was to evaluate the feasibility of sentinel lymph node biopsy in breast cancer patients at our institution and to compare the results of sentinel node (SN) biopsy to standard axillary lymph-node dissection (ALND). METHODS: In a retrospective study the percentage of lymph-node positive patients and the number of micrometastases in 165 breast cancer patients following SN biopsy was compared to 195 patients who underwent ALND of level I and II without SN biopsy. The SN was identified using a combination of vital blue dye and a radiolabelled colloid. RESULTS: Patients and tumour characteristics were comparable between both groups. SN biopsy found no significant difference in the number of node positive T1 cancer patients (SN group: 31/108 (28.7%) -- ALND group: 21/92 (22.8%)) and T2 tumours (SN group: 27/57 (47.4%) -- ALND group: 49/103 (47.6%)) between both groups. Micrometastases were more frequently found in the SN group when compared to the ALND group (six of 70 positive nodes) (P=0.04). CONCLUSION: SN biopsy may be as accurate as standard axillary lymph node dissection for the evaluation of the axillary lymph-node status in breast cancer patients. PMID- 11417985 TI - Parathyroid carcinoma: problems in diagnosis and the need for radical surgery even in recurrent disease. AB - INTRODUCTION: From 1986 to 1999 we operated on 963 patients with primary hyperparathyroidism (pHPT). METHODS AND RESULTS: Parathyroid carcinoma was diagnosed clinically and histologically in four patients (0.4%). In two of these patients diagnosis of parathyroid cancer was delayed by misinterpretation of the histopathology leading to an autotransplantation of malignant parathyroid tissue in one case. In two patients multivisceral surgery was performed: beside thyroidectomy, neck dissection, tracheal wall resection and resection of the muscular layer of the oesophagus one patient received oesophagectomy and gastric transposition and one patient a lung wedge resection. Both patients had a temporary palliation of tumour-associated symptoms after multivisceral surgery. The first patient died 2 years after oesophagectomy and 12 years after primary diagnosis from local tumour recurrence and cachexia. The second patient is living with tumour recurrence presenting a serum calcium level of 4.2 mmol/l (normal range 2.0 to 2.5 mol/l) and multiple brown tumours 2 years after lung resection and 6 years after the primary diagnosis. CONCLUSIONS: We conclude that parathyroid carcinomas, being difficult to diagnose, warrant radical surgery, including multivisceral resection to prolong survival and reduce tumour and hypercalcaemia associated symptoms. PMID- 11417986 TI - Regional toxicity after isolated limb perfusion with melphalan and tumour necrosis factor- alpha versus toxicity after melphalan alone. AB - AIMS: To determine whether the addition of high-dose tumour necrosis factor-alpha (TNF alpha) to isolated limb perfusion (ILP) with melphalan increases acute regional tissue toxicity compared to ILP with melphalan alone. METHODS: A retrospective, multivariate analysis of toxicity after normothermic (37--38 degrees C) and 'mild' hyperthermic (38--40 degrees C) ILPs for melanoma was undertaken. Normothermic ILP with melphalan was performed in 294 patients (70.8%), 'mild' hyperthermic ILP with melphalan in 71 patients (17.1%) and 'mild' hyperthermic ILP with melphalan combined with TNF alpha in 50 patients (12.0%). Toxicity was nil or mild (grades I--II according to Wieberdink et al.) in 339 patients (81.7%), and more severe acute regional toxicity (grades III--V) developed in 76 patients (18.3%). A stepwise logistic regression procedure was performed for the multivariate analysis of prognostic factors for more severe toxicity. RESULTS: On univariate analysis, 'mild' hyperthermic ILP with melphalan plus TNF alpha significantly increased the incidence of more severe acute regional toxicity compared to normothermic and 'mild' hyperthermic ILP with melphalan alone (36% vs 16% and 17%; P=0.0038). However, after ILP using TNF alpha no grade IV (compartment compression syndrome) or grade V (toxicity necessitating amputation) reactions were seen. Significantly more severe toxicity was seen after ILPs performed between 1991 and 1994 compared with earlier ILPs (33%vs 14%P=0.0001). Also, women had a higher risk of more severe toxicity than men (22% vs 7%; P=0.0007). After multivariate analysis, prognostic factors which remained significant were: sex (P=0.0013) and either ILP schedule (P=0.013) or treatment period (P=0.0003). CONCLUSIONS: Regional toxicity after 'mild' hyperthermic ILP with melphalan and TNF alpha was significantly increased compared to ILP with melphalan alone. This may be caused by increased thermal enhancement of melphalan due to the higher tissue temperatures (39--40 degrees C) at which the melphalan in the TNF alpha-ILPs was administered or by an interaction between high-dose TNF alpha and melphalan. PMID- 11417987 TI - Activated human neutrophils release hepatocyte growth factor/scatter factor. AB - BACKGROUND: Hepatocyte growth factor or scatter factor (HGF/SF) is a pleiotropic cytokine that has potent angiogenic properties. We have previously demonstrated that neutrophils (PMN) are directly angiogenic by releasing vascular endothelial growth factor (VEGF). We hypothesized that the acute inflammatory response can stimulate PMN to release HGF. AIMS: To examine the effects of inflammatory mediators on PMN HGF release and the effect of recombinant human HGF (rhHGF) on PMN adhesion receptor expression and PMN VEGF release. METHODS: In the first experiment, PMN were isolated from healthy volunteers and stimulated with tumour necrosis factor-alpha (TNF-alpha), lipopolysaccharide (LPS), interleukin-8 (IL 8), and formyl methionyl-leucyl-phenylalanine (fMLP). Culture supernatants were assayed for HGF using ELISA. In the second experiment, PMN were lysed to measure total HGF release and HGF expression in the PMN was detected by Western immunoblotting. Finally, PMN were stimulated with rhHGF. PMN CD 11a, CD 11b, and CD 18 receptor expression and VEGF release was measured using flow cytometry and ELISA respectively. RESULTS: TNF-alpha, LPS and fMLP stimulation resulted in significantly increased release of PMN HGF (755+/-216, 484+/-221 and 565+/-278 pg/ml, respectively) compared to controls (118+/-42 pg/ml). IL-8 had no effect. Total HGF release following cell lysis and Western blot suggests that HGF is released from intracellular stores. Recombinant human HGF did not alter PMN adhesion receptor expression and had no effect on PMN VEGF release. CONCLUSIONS: This study demonstrates that pro-inflammatory mediators can stimulate HGF release from a PMN intracellular store and that activated PMN in addition to secreting VEGF have further angiogenic potential by releasing HGF. PMID- 11417988 TI - (131)I-MIBG radionuclide therapy is safe and cost-effective in the control of symptoms of the carcinoid syndrome. AB - BACKGROUND: The standard treatment used to control the symptoms of carcinoid syndrome (CS) involves subcutaneous injections of the somatostatin analogue octreotide. This is expensive (US $8000--16,000 per year), and treatment may be for many years. The aim of this study was to evaluate the efficacy and cost effectiveness of our experience over the last 5 years with 1-131-labelled metaiodobenzylguanidine (MIBG) radionuclide therapy in the palliation of patients with CS. METHODS: A consecutive series of 20 symptomatic patients (referred between 1994 and 1999) with CS were evaluated. Fifteen of them underwent(123)I MIBG scanning. Of the 13 patients with significant tracer uptake in metastatic deposits compared to background, 12 underwent a course of therapeutic(131)I-MIBG (one patient refused). Symptoms, biochemical markers, CT scans, follow-up(123)I MIBG scans, and the requirement for octreotide were used to assess outcome of treatment. Costs of(131)I-MIBG and octreotide treatments were compared. RESULTS: MIBG treatment was well tolerated in all with only transient side-effects. Ten patients showed a measurable clinical improvement. Seven had a complete clinical response. The mean duration of response was 15.4 months. Octreotide was not required or was reduced in eight patients. Treatment with(131)I-MIBG resulted in a saving of US $1000 per patient, with effective symptom control, when compared to octreotide. CONCLUSION: 1-131 MIBG therapy is a safe and cost-effective therapeutic option to successfully control symptoms in patients with carcinoid syndrome. PMID- 11417989 TI - Incidence and clinical course of de-novo malignancies in renal allograft recipients. AB - BACKGROUND: This study was designed to evaluate the development of malignancies after renal transplantation in a single centre. The outcome was studied in patients developing a malignant neoplasm after renal transplantation. METHODS: Malignancies are registered in a database containing relevant data about the patients with a renal transplant. This database and the files of the patients developing a malignant neoplasm, have been studied as to stage at presentation, therapy and outcome. RESULTS: In 1546 patients with 2075 renal transplantations, 240 malignancies developed in 231 recipients. Skin cancers often present with more than one lesion of the same histological type. After the first skin tumour, about half of the patients developed more lesions, of the same or a different histological type. The prognosis of skin tumours is relatively good, but most malignancies in all other categories have a poor prognosis. CONCLUSIONS: Cutaneous neoplasms tend to be multiple, but can be controlled by regular examination of the skin. Most malignant lymphomas do develop outside the lymphoproliferative system and have a poor prognosis. Patients with a solid tumour of the other tracts often present in an advanced stage of disease, which makes the outcome of treatment, if possible at all, disappointing. PMID- 11417990 TI - Technical details for the sentinel node biopsy in breast cancer: a guide for the training process. PMID- 11417991 TI - Dismiss low back pain in renal cell carcinoma patients at your peril: meningeal cauda equina deposits. PMID- 11417992 TI - Male breast neoplasia in association with selective serotonin re-uptake inhibitor therapy: a report of three cases. AB - Male breast cancer is a rare condition with very poorly understood risk factors. We report three cases of men with malignant and pre-malignant breast disease who had all been prescribed selective serotonin re-uptake inhibitors (SSRIs) for depression. Concerns about an association between this group of drugs and breast cancer in women have been previously raised and experimental evidence has suggested that these drugs could influence regulation of cellular proliferation acting through internal cellular messengers. Risk factors for the development of breast cancer are likely to be multifactorial, possibly more so in women given the complex physiological changes that occur in the female breast. Whilst the cases we report are anecdotal and other risk factors may be present, we suggest that assessment of any possible contribution that SSRI therapy may make to the development of breast neoplasia may be more easily assessed in a male population. PMID- 11417993 TI - Metastatic malignant acrospiroma of the hand. AB - We present the case of a 37-year-old man with multiple pulmonary metastases of a primarily unknown primary tumour. Thorough revision of the medical history yielded that he had already passed three resections of a right palmar mass, which had been described as a benign tumour. Clinical examination showed a thickened scar with a suspicious palpable mass in the right hand. Excision of this scar and the tumour mass with histopathological examination now revealed a malignant acrospiroma. Resection of the pulmonary metastases histologically also confirmed a malignant acrospiroma. The following radical resection of the metacarpals II and III with the index and middle finger under the assumption of a wide compartment resection achieved tumour free margins and proved to be efficient with the patient being relapse free for 4 years from this operation. Although the prognosis of this tumour is generally unfavourable this particular case demonstrates the value of a radical surgical resection as the mainstay of treating such highly malignant sweat gland tumours of the hand. PMID- 11417994 TI - Choriocarcinomatous metaplasia of a metachronous adenocarcinoma of the colon. AB - A second primary colonic adenocarcinoma developed in a 68-year-old man following resection of a rectal adenocarcinoma. Choriocarcinomatous change was found in the metachronous lesion with liver metastases and elevated beta-human choriogonadotrophin (HCG) serum titres. PMID- 11417996 TI - Heterogeneity of so-called neuroendocrine lung tumors. AB - The diagnosis of neuroendocrine (NE) lung tumor is dependent on a number of observations: organoid structure, dense core granules, and various molecular components, including chromogranin A, neurosecretory enolase, synaptophysin, neural cell adhesion molecules, and others. None of these is specific for lung tumors. The Kulchitsky cell, which has these characteristics, forms a carcinoid, which exemplifies the NE tumor. It is euploid, has few mitoses, no necrosis and a 5- to 10-year survival of over 90%. When carcinoids show malignant characteristics, i.e., increased mitoses and necrosis, they have been labeled atypical and have a survival of 50%. Because all other non-small cell lung tumors, especially large cell tumors, may show one or more of these things because of the inherent heterogeneity of lung tumors, the term NE has been applied to them without real evidence that this affects survival with or without chemotherapy. This is expensive and without clinical significance. PMID- 11417997 TI - Cooperating oncogenic events in murine mammary tumorigenesis: assessment of ErbB2, mutant p53, and mouse mammary tumor virus. AB - We are investigating cooperating genetic events in the genesis of breast cancer, using the mouse as a model system. We have shown cooperativity between a mutant allele of p53 (p53-172H) and overexpressed ErbB2 in mammary tumorigenesis in transgenic mice. We are now performing additional crosses to further examine oncogene cooperativity with ErbB2 and p53-172H. We attempted to test the dominant oncogenic potential of p53-172H in an in vivo setting by crossing the p53-172H transgene together with ErbB2 onto either a p53(-/-) or a p53(+/-) background. We show that the p53-172H allele and the heterozygous p53 genotype have an identical impact on the latency of ErbB2-induced mammary tumors; there was no evidence of additivity or synergy between p53-172H and the p53(+/-) genotype. On the p53(-/-) background, we obtained no mammary tumors due to the early onset of lymphomas and sarcomas, thus precluding assessment of the effect of the p53-172H transgene on mammary tumorigenesis in a p53-null background. Thus, in this in vivo model for breast cancer, we failed to find evidence that p53-172H can function as a dominant oncogenic allele, but rather found support for its being essentially equivalent to a null allele in its impact on ErbB2-induced mammary tumorigenesis. By comparative genome analysis, we showed that a common feature of tumors arising in ErbB2/mutant p53 mice (p53-null allele with or without p53-172H) is a loss of chromosome 4, a feature of many epithelial tumors in mice and one that is consistent with a role for loss of INK4a/ARF in such tumors. We also attempted to accelerate ErbB2-induced mammary tumorigenesis with mouse mammary tumor virus (MMTV) proviral tagging mutagenesis, but we were surprised to find that mice with MMTV alone had the same latency as mice with both MMTV and ErbB2, indicating no cooperativity between ErbB2 and MMTV. This may have been due to the mixed C3H/HeN x FVB strain background used in this cross. PMID- 11417998 TI - Rapid, efficient genotyping of clinical tumor samples by laser-capture microdissection/PCR/SSCP. AB - BACKGROUND: Mutation analysis is becoming increasingly important in clinical practice, since sporadic mutations in tumors often correlate with prognosis and/or therapeutic response. However, the labor-intensive nature of the molecular analyses has limited the routine clinical use of tumor genotyping. Laser-capture microdissection (LCM) allows procurement of relatively pure tumor cell populations. We have investigated the possibility that the use of laser-capture microdissection would allow elimination of time-consuming intermediate steps in tumor genotyping. Design. Archival formalin-fixed, paraffin-embedded tissues from seven cases of colorectal adenocarcinoma were laser- and hand-microdissected and subsequently evaluated by PCR/SSCP/sequencing for Ki-ras exon 1 and p53 exons 5, 7, and 8. Results. Mutations in Ki-ras exon 1 and/or p53 exons 5 and 7 were detected in five of the seven samples. In the hand-microdissected samples, confident identification of mutations was possible in several cases only after band excision, DNA elution, reamplification, and verification of mutant enrichment by a second SSCP analysis prior to sequencing. In the laser microdissected samples, confident mutation identification was possible in all cases with direct sequencing of the original PCR product, reducing the time required for molecular analysis to 3 days. Conclusion. Using laser-capture microdissection, mutant signals are strong enough to sequence directly from original PCR products. With rapid, efficient genotyping by LCM/PCR/SSCP, results can be incorporated directly into the surgical pathology report. PMID- 11417999 TI - pp60(c-src) modulates microvascular endothelial phenotype and in vitro angiogenesis. AB - The tyrosine kinase c-src associates with the platelet-derived growth factor (PDGF) receptor. Overexpression of wild-type c-src, a kinase-negative c-src mutant, and v-src in microvascular endothelial cells modulated the mitogenic effect of PDGF, suggesting that c-src kinase activity inhibits PDGF signals. Analyses of cell morphology in two-dimensional culture revealed changes in cell shape and size induced by the overexpression of c-src proteins. Investigations in three-dimensional culture unveiled a modulatory role of c-src during in vitro angiogenesis. Overexpression of c-src resulted in an increased diameter of tube like structures, and the number of branching segments was decreased. Expression of the kinase-negative c-src mutant resulted in abortive tube formation consisting of disconnected multicellular fragments. These results indicate that the c-src tyrosine kinase exerts regulatory effects on endothelial proliferation, size, and cytoskeletal organization in two-dimensional culture and on the formation of a differentiated multicellular network in three-dimensional culture. PMID- 11418000 TI - Spectrin oligomerization is cooperatively coupled to membrane assembly: a linkage targeted by many hereditary hemolytic anemias? AB - In the erythrocyte, ankyrin is the major adapter protein linking tetramers of band 3 to the spectrin-actin cytoskeleton. This linkage involves a direct interaction between ankyrin and the 14th-15th repeat unit of beta-spectrin. The spectrin cytoskeleton itself is stabilized by the self-association of spectrin heterodimers into tetramers and larger oligomers, a process mediated by the 17th repeat unit of beta-spectrin and a short NH(2) -terminal sequence in alpha spectrin. The self-association of spectrin and its ankyrin-mediated membrane binding have generally been considered independent events. We now demonstrate that spectrin self-association, the binding of spectrin to ankyrin, and the binding of ankyrin to the 43-kDa cytoplasmic domain of band 3 (cdb3) are coupled in a positively cooperative way. In solution, [(125)I]-labeled ankyrin was found by ND-PAGE3 to enhance the affinity of spectrin self-association by 10-fold. The reciprocal process was also true, in that spectrin tetramers and oligomers bound ankyrin with enhanced affinity relative to dimer spectrin. Saturation of the beta spectrin self-association site by an NH(2) -terminal 80-kDa alpha-spectrin peptide enhanced the affinity of spectrin dimer for ankyrin, indicating a direct relationship between ankyrin binding and the occupancy of the beta-spectrin self association site. cdb3 accentuated these cooperative interactions. Several inherited spectrin mutations that cause hemolytic disease but that do not directly destabilize the self-association or ankyrin-binding sites can be explained by these results. Three classes of mutations appear to disrupt cooperative coupling between self-association and ankyrin binding: (i) mutation of the linker sequences that join helices C and A in repeat units that intervene between the two functional sites, mutations that presumably block repeat-to repeat transfer of conformational information; (ii) mutations in alpha-spectrin repeats 4 to 6 that disrupt the ability of this region to trans-regulate ankyrin binding by the adjacent beta-spectrin repeats 14-15; and (iii) exon-skipping mutations that shorten alpha-spectrin and force repeats 4 to 6 to fall out-of register with the ankyrin-binding motif in beta-spectrin. Collectively, these results demonstrate a molecular mechanism whereby a membrane receptor can directly promote cytoskeletal assembly. PMID- 11418001 TI - Dysfunction of Stat4 leads to accelerated incidence of chemical-induced thymic lymphomas in mice. AB - Stat4 (signal transducer and activator of transcription) can be activated by specific cytokines, such as IL-12, IFN-alpha, and IL-2. Since IL-12 has been implicated in tumor surveillance and cancer treatment, we hypothesized that its signaling mediator, Stat4, may repress tumor growth. Mice lacking Stat4 allowed us to directly assess the role of Stat4 in tumor surveillance. Lymphomas were chemically induced by MNU (N-methyl-N-nitrosourea) injection in Stat4-deficient or wild-type control mice. At the time of termination of the experiment 16 weeks after injection, 78% of homozygous Stat4-deficient mice had developed thymic lymphomas. This tumor induction was dramatically higher than in heterozygous (14%) and wild-type controls (14%). Lymphoma development occurred 5 weeks earlier in homozygous knockout mice than in other genotypes. Mice bearing tumors were fragile and had an increased death rate in the early stages of the experiment. The tumors displayed a very aggressive phenotype with metastases in multiple organs. Therefore, the loss of Stat4 predisposes mice to tumor induction and demonstrates crucial roles of Stat4 in the prevention of tumors. PMID- 11418002 TI - Regulation of MCP-3 and BRCA2 mRNA expression levels by beta(1) integrins. AB - The integrin cytoplasmic domain has been shown to modulate several cellular functions, including cell proliferation, adhesion, migration, and intracellular signaling. The beta(1) integrin subunits beta(1C) and beta(1A), which contain variant cytoplasmic domains, differentially affect cancer and normal cell functions. To identify target genes selectively regulated by these beta(1) cytoplasmic variants, stable cell transfectants expressing either beta(1A) or beta(1C) under the control of a doxycycline-inducible promoter were obtained using murine beta(1)-deficient GD25 cells. Screening of 1176 murine cDNAs using first-strand cDNA of mRNA isolated from either beta(1C)- or beta(1A)-expressing cells showed a striking differential expression of few genes. The differential expression of two genes, MCP-3 and BRCA2 (monocyte chemoattractant protein-3 and breast cancer susceptibility gene 2, respectively), whose products are involved, respectively, in chemotaxis and embryonic proliferation, was confirmed by Northern blot analysis. Increased MCP-3 and decreased BRCA2 mRNA levels in cells expressing beta(1C) compared to those in cells expressing beta(1A) were observed. Since beta(1C) and beta(1A) stable cell transfectants showed comparable adhesion to fibronectin, upregulation of MCP-3 and downregulation of BRCA2 mRNA levels did not appear to be due to a differential ability of the beta(1C) cells to adhere to the beta(1) ligand fibronectin. Overall, our data show that beta(1) integrin cytoplasmic domain variants control expression of downstream target genes in a differential manner without affecting cell adhesion. PMID- 11418003 TI - Expression of the apoptosis inhibitor survivin in aggressive squamous cell carcinoma. AB - Deregulated expression of inhibitors of apoptosis (programmed cell death) may contribute to cancer by aberrantly extending cell viability and facilitating the insurgence of resistance to therapy. In this study, we investigated the potential expression and prognostic significance of the apoptosis inhibitor survivin in squamous cell carcinoma (SCC). A series of 135 cases of SCC including 46 oral SCC and 89 cutaneous SCC was analyzed for survivin expression by immunohistochemistry and Western blotting. Survivin was found in 57 cases (64%) of skin SCC and 26 cases (56%) of oral SCC, with weighted survivin scores ranging from 1 to 12. In contrast, normal oral epithelium, normal skin epithelium, and skin annexa did not express survivin. Survivin expression significantly (P < 0.05) segregated with high-grade and undifferentiated tumors with size >1.5 cm and invariably associated with lymph node metastasis. These data suggest that survivin expression may predictively identify cases of SCC with more aggressive and invasive clinical phenotype, potentially warranting closer follow-up protocols. PMID- 11418004 TI - Amplification of tissue by construction of tissue microarrays. AB - Tissue microarrays are a method of relocating tissue from conventional histologic paraffin blocks in a manner that tissue from multiple patients or blocks can be seen on the same slide. This is done by using a needle to biopsy a standard histologic section and placing the core into an array on a recipient paraffin block. This technique allows maximization of tissue resources by analysis of small core biopsies of blocks, rather than complete sections. Using this technology, a carefully planned array can be constructed using cases from pathology tissue block archives, and a 20-year survival analysis can be done on a cohort of 600 or more patients using only a few microliters of antibody in a single experiment. Furthermore, this cohort can be analyzed thousands of times with different reagents as a result of judicious sectioning of the array block. This review describes this process and discusses the issues of representative sampling in heterogeneous lesions, the issue of antigen preservation, and some technical strategies and methods of array construction. In summary, this technique can provide a highly efficient, high-throughput mechanism for evaluation of protein expression in large cohorts. It has the potential for allowing validation of new genes at a speed comparable to the rapid rate of gene discovery afforded by DNA microarrays. PMID- 11418005 TI - Differential activation of the STAT pathway by angiotensin II via angiotensin type 1 and type 2 receptors in cultured human fetal mesangial cells. AB - The vasoactive peptide angiotensin II is the principal effector of the renin angiotensin system. It exerts mitogenic and growth-inhibiting effects in many target tissues, including renal mesangial cells. To investigate mechanisms of angiotensin II signaling in human mesangial cells, we explored the signal transducer and activator of transcription (STAT) pathway as a possible regulator of angiotensin II receptor-specific signaling. We tested whether angiotensin II could induce STAT activation and nuclear translocation of STAT proteins in human mesangial cells by electromobility shift assays and by immunostaining and confocal microscopy. We found that fetal human mesangial cells express STAT1,2,3,5, and 6 and that stimulation of these cells by angiotensin II results in rapid induction of STAT1 and STAT5 DNA-binding activity. This DNA-binding activity was identified as STAT5 for angiotensin receptor type 1 activation and STAT1 for angiotensin receptor type 2-mediated activation, as induction of STAT DNA binding by angiotensin II could be differentially blocked by the angiotensin receptor type 1 blocker losartan and by angiotensin II receptor type 2 blocker PD 123,319. Angiotensin II also induced STAT1 and STAT5 tyrosine phosphorylation and nuclear translocation of activated STATs in a receptor subtype-specific manner. STAT activation thus appears to provide an important signaling pathway for angiotensin II-induced cellular responses. PMID- 11418006 TI - Epitope-specific antibodies to the beta(1C) integrin cytoplasmic domain variant. AB - The beta(1C) integrin is an alternatively spliced variant of the beta(1) subunit that contains a unique 48-amino-acid sequence in its cytoplasmic domain. We have shown previously that beta(1C) is a potent inhibitor of cell proliferation and that in vivo its expression is downregulated in prostate and breast carcinoma. In this study, we describe a panel of specific monoclonal antibodies that react with the beta(1C) cytodomain. We show by immunoblot analysis that the newly generated monoclonal antibodies specifically recognize the beta(1C) cytodomain expressed as glutathione S-transferase fusion protein. The specificity of the antibodies to beta(1C) was confirmed in competition studies by immunoblotting using beta(1C) specific synthetic peptides. These monoclonal antibodies reacted, in enzyme linked immunosorbent assays, with the beta(1C) 785-808 peptide but failed to bind the beta(1C) 778-794, beta(1C) 805-825, or beta(1A) 765-798 peptides. Thus, the epitope recognized by the antibodies is located within the Q(795)-F(804) beta(1C) cytoplasmic sequence; this region overlaps the previously described Q(795)-Q(802) domain necessary for beta(1C) to inhibit cell proliferation. To our knowledge, these are the first monoclonal antibodies specific for a beta(1) cytoplasmic isoform. The monoclonal antibodies described here will be useful tools for dissecting functional differences, among beta(1) integrin variants, as well as for the study of the role of beta(1C) in prostate and breast epithelial cell proliferation. PMID- 11418007 TI - In situ detection of messenger RNA using digoxigenin-labeled oligonucleotides and rolling circle amplification. AB - The detection of specific RNA molecules in situ is routinely performed using haptenated probes, which are detected by either enzymatic amplification or direct fluorescence. A drawback of fluorescence labeling has been the reduced sensitivity relative to that of methods that use enzymes as signal generators. Reliable fluorescence detection methods often require the use of multiple oligonucleotide probes for each gene target. Here, we demonstrate that single haptenated DNA probes specific for actin mRNA may be detected in situ using antibody-coupled rolling circle amplification (immuno-RCA). This fluorescence based detection method offers remarkable sensitivity due to the use of signal amplification and yet retains the ability to count hybridization signals as discrete objects. We demonstrate the detection of actin-specific immuno-RCA signals in the cytoplasm and use 3D image deconvolution of multiple z axis sections to show that there are hundreds of signals per cell. With some modifications, this method may be adaptable to the simultaneous detection of several RNA species, including low-copy-number mRNA. PMID- 11418008 TI - Detection of the mutated K-Ras biomarker in colorectal carcinoma. AB - This study describes an advantageous, effective protocol for detecting K-Ras mutations in human stool as a prototype screen for colorectal carcinoma (CRC), the third most common malignancy in the United States. A reliable screening test that detects early lesions would contribute to a decrease in mortality. Currently, the only noninvasive screen for CRC is the hemeoccult, test which has a high false-positive rate. Previously, several investigators have identified genetic biomarkers for CRC in stool DNA. The K-Ras oncogene, mutated in 46-50% of CRC tumors, serves as one molecular marker by which stool samples may be evaluated for early detection of adenocarcinomas. DNA was isolated from stool samples by a new method we specifically designed for extracting high-quality DNA using tetradecyltrimethylammonium oxalate [Catrimox-14, Iowa Biotechnology Corp., (currently Qiagen)]. This protocol produces an optimal yield of high-purity DNA, suitable for genotyping. Detection of the human gene in stool samples was enhanced by hybrid selection of the K-Ras sequences, polymerase chain reaction, and single-strand conformation polymorphism. Tumor tissue and preoperative stool samples for eight patients were K-Ras genotyped and compared; stool samples from two asymptomatic, healthy patients were also evaluated in a double-blind format. In seven of eight samples (87%), the genotypes of the stool and colon tissue DNA were the same. Both healthy patients showed wild-type K-Ras. This protocol shows promise for the development of an efficient and accurate screen for CRC. PMID- 11418009 TI - Pathobiologic findings in DCIS of the breast: morphologic features, angiogenesis, HER-2/neu and hormone receptors. AB - With the increasing incidence of ductal carcinoma in situ (DCIS) of the breast and its relationship to invasive breast carcinoma, it is important to understand the biology of this entity. We report on a hospital-based survey of 219 case subjects with DCIS of the breast without associated invasive carcinoma diagnosed between 1982 and 1994. The cases of DCIS were analyzed for architectural type, size, nuclear grade, necrosis, calcification, periductal fibrosis, neovascularization, estrogen receptor (ER), progesterone receptor (PR), and HER 2/neu expression. Periductal neovascularization was associated with tumor size, microcalcifications, periductal fibrosis, and HER-2/neu overexpression. Expression of ER and PR was observed in 60 and 62% of the cases, respectively, and HER-2/neu overexpression in 28% of the cases. ER and PR expression were both inversely associated with comedo histology and nuclear grade. HER-2/neu overexpression was positively associated with comedo histology, high nuclear grade, and periductal neovascularization and was inversely correlated with both ER and PR expression. High nuclear grade was positively associated with comedocarcinoma, necrosis, microcalcification, and periductal fibrosis. The role of these molecular/pathologic markers in the biology of DCIS and their potential clinical implications are discussed. PMID- 11418010 TI - TNF signaling in vascular endothelial cells. AB - Vascular endothelium is a major target of actions of the proinflammatory cytokine tumor necrosis factor (TNF). Increasingly, the intracellular pathways that are activated in response to TNF have been elucidated. Many of these pathways have proven to be cell type-specific, requiring that observations made in other cell types be confirmed or ruled out in endothelial cells (EC). In this review the authors will summarize the state of the field, emphasizing studies in cultured human EC. PMID- 11418011 TI - Phosphoproteomics. PMID- 11418012 TI - Factors affecting autopsy rates, autopsy request rates, and autopsy findings at a large academic medical center. AB - Autopsy rates continue to decline in the United States. To assess the impact of various objective factors (time of death, day of death, age at death, patient gender, clinical service, and length of hospital stay) on the autopsy request rate, autopsy rate, successful request rate, and percentage of cases in which the autopsy examination added to or altered the clinical assessment of the patient, we prospectively studied all hospital deaths at a major academic medical center for the 3-year period from 1996 through 1998. The autopsy rate decreases significantly with patient age, both because of a decreasing request rate and because family members are less likely to grant permission. An autopsy is less likely to be requested for deaths in the emergency department or on general surgery services and most likely to be requested for fetal, medicine, cardiothoracic surgery, and pediatric deaths. Families more commonly grant permission for autopsy on fetal deaths, pediatric deaths, and emergency department deaths. Forty percent of autopsies reveal significant information about the patient's death beyond what was known premortem. This is least frequent among the fetal deaths, but relatively constant for adults of all ages. Patients who die in the emergency department are most likely to have significant unexpected findings at autopsy. Increasing the request rate for adult patients who die in the emergency department and on the medicine services will result in the greatest increase in information learned from autopsy. PMID- 11418013 TI - Pathology of epilepsy. PMID- 11418014 TI - Evaluation of classical NSAIDs and COX-2 selective inhibitors on purified ovine enzymes and human whole blood. AB - Cyclooxygenase is the key enzyme in the biosynthesis of prostanoids, biologically active substances involved in several physiological processes and also in pathological conditions such as inflammation. It has been well known for 10 years that this enzyme exists under two forms: a constitutive (COX-1) and an inducible form (COX-2). Both enzymes are sensitive to inhibition by conventional non steroidal anti-inflammatory drugs (NSAIDs). Observations were made that COX-1 was mainly involved in homeostatic processes, while the COX-2 expression was associated with pathological conditions leading to the development of COX-2 selective inhibitors. Several methods have been reported for the evaluation of the COX-1 and COX-2 inhibitory potency and selectivity of conventional or COX-2 selective NSAIDs. In this study, we evaluated the COXs inhibitory profile of both conventional NSAIDs and COX-2 selective inhibitors using two different in vitro methods: the first test was performed using purified enzymes while the second method consisted of a whole blood assay. The results obtained with reference drugs in these two assays will be discussed and compared in this article. PMID- 11418015 TI - Evaluation of the activity and localization of 15-lipoxygenase-1 after introduction into human colorectal carcinoma Caco-2 cells. AB - In human colorectal carcinoma Caco-2 cells, sodium butyrate (NaBT) induces the expression of the reticulocyte, 15-lipoxygenase-1 (15-LO-1) and causes these cells to undergo differentiation and apoptosis. 15-LO-1 is also expressed in human colorectal epithelium with a significant higher expression observed in colorectal tumors. In this study, we have prepared stable Caco-2 cells that expressed 15-LO-1 under control of an inducible promoter. These cells provide a model system to study regulation of 15-LO-1 activity in colorectal cells without the interfering presence of NaBT and are useful to study the biological function of 15-LO-1. The expressed 15-LO-1 was highly active as measured in cell lysates, but we were unable to detect metabolism in intact cells. The addition of calcium to the media for the Caco-2 cells was required for 15-LO-1 to translocate from the cytosol to the membrane which is frequently a requirement for lipoxygenase activity. Despite the addition of calcium and translocation, little lipoxygenase activity was detected with intact cells. However, after removal of phenol red, a common constituent of cell culture media, we were able to detect 15-LO-1 activity in the transfected Caco-2 cultured cells. Thus the presence of calcium and the absence of antioxidants present in commonly used culture media are required for expressed 15-LO-1 to be catalytically active and to permit an examination of its biological effects. PMID- 11418016 TI - Arachidonyl trifluoromethyl ketone induces lipid body formation in leukocytes. AB - Leukocyte lipid bodies, abundant in cells associated with inflammation, can be induced to form in response to stimuli that include cis -unsaturated, but not saturated, fatty acids. Arachidonyl trifluoromethyl ketone (AACOCF(3)), a non esterifiable arachidonate analog and an inhibitor of cytosolic phospholipase A(2)enzymes (PLA(2)), dose-dependently (0-20 microM) stimulated neutrophil lipid body formation, but this stimulation was not attributable to PLA(2)inhibition. Palmitoyl trifluoromethyl ketone, also a PLA(2)inhibitor, failed to stimulate lipid body formation, like palmitic acid itself, and did not inhibit stimulated lipid body formation. Moreover, aspirin, indomethacin and ibuprofen, which inhibit cis -unsaturated fatty acid-induced lipid body formation, inhibited AACOCF(3)-induced lipid body formation. Lipid body induction with AACOCF(3)reflected its structural basis as a cis -unsaturated fatty acid analog. These results indicate that cytosolic PLA(2)enzymes are not active in lipid body induction and cis -fatty acid stimulation of lipid body formation does not require esterification of cis -fatty acids into glycerolipids. PMID- 11418017 TI - Role of prostanoids and endothelins in the prevention of cyclosporine-induced nephrotoxicity. AB - Cyclosporine A nephrotoxicity includes both functional toxicity and histological changes, whose seriousness is dependent upon the dose and the duration of the drug administration. Several vasoactive agents have been found to be implicated in cyclosporine induced nephrotoxicity, among which prostanoids and endothelins are the most important. In previous studies we were able to prevent the early stage (7 days) of cyclosporine (37.4 micromol [45 mg]/kg/day) induced nephrotoxicity in rats either by the administration, i) of OKY-046, a thromboxane A(2)synthase inhibitor, ii) of ketanserine, an antagonist of S(2)serotonergic, a(1)adrenergic, and H(1)histaminergic receptors and iii) of nifedipine, a calcium channel blocker, or by diet supplementation either with evening primrose oil or fish oil. All these protective agents elevated ratios of excreted renal prostanoid vasodilators (prostaglandins E(2), 6ketoF(1 alpha)) to vasoconstrictor (thromboxane B(2)), a ratio which was decreased by the administration of cyclosporine alone. Nifedipine averted the cyclosporine induced increase of urinary endothelin-1 release. All protections were associated with the reinstatement of glomerular filtration rate forwards normal levels whereas renal damage defence, consisting of a decrease of the cyclosporine induced vacuolizations, was variable. Ketanserine and evening primrose oil were the only agents which prevented the animal body weight loss. These data suggest that prostanoids and endothelin-1 may mediate functional toxicity while thromboxane A(2)is involved the morphological changes too, provoked in the early stage of cyclosporine treatment. However, other nephrotoxic factors and additional mechanisms could also be implicated in the cyclosporine induced nephrotoxicity. PMID- 11418018 TI - Peroxisome proliferator-activated receptor- gamma expression in human malignant and normal brain, breast and prostate-derived cells. AB - The constitutive and gamma -linolenic acid (GLA)-induced expression of peroxisome proliferator-activated receptor gamma (PPAR gamma) immunoreactive protein in a panel of human malignant brain (U87MG, T98G); breast (MCF-7, MB MDA-231, MB MDA 435) and prostate (ALVA, DU-145, LNCaP, PC3) cell lines have been compared with those for their normal cell counterparts, the human normal astrocyte (NHA), mammary epithelial (HMEC) and prostate epithelial (PrEC) cells, respectively. Constitutive levels of expression for PPAR gamma protein were significantly higher in the malignant cell lines relative to their normal cells. GLA supplementation did not affect the protein expression in malignant cells but caused 6- and 3-fold increases in normal breast and prostate cells, respectively. Since activation of PPAR gamma protein in some human malignant cell lines has been demonstrated to induce tumour cell death, these findings signal the need to exploit the significantly elevated expression of this protein in the therapy of human cancer. PMID- 11418019 TI - Cytosolic phospholipase A(2)and 15-hydroxyprostaglandin dehydrogenase mRNA expression in murine uterine and gestational tissues during late pregnancy. AB - We evaluated the changes in mRNA expression of cytosolic phospholipase A(2)(cPLA(2)) and 15-hydroxyprostaglandin dehydrogenase (PGDH) in intrauterine and gestational tissues during mid-late murine pregnancy. Tissues (decidual caps, fetal membranes, and placentae, uterus, and cervix) were collected from pregnant mice at days 12, 14, 16, 18, and 19 (am and pm) of gestation. Total RNA was isolated and evaluated for cPLA(2)and PGDH expression by northern blot analysis normalized to GAPDH expression. Expression of mRNA for cPLA(2)increased in the placentae and decidual caps on day 18 and 19 pm, respectively. There was also increased expression for PGDH mRNA in the placenta and fetal membranes at the later stages of pregnancy. The tissue specific differences in expression of cPLA(2)and PGDH suggest that changes in enzymatic regulation of PG production and degradation may be crucial for the initiation of labour. PMID- 11418020 TI - Nitric oxide and endothelin relationship in intestinal ischemia/reperfusion injury (II). AB - Endothelins ( ETs ) are potent vasoconstrictors derived from vascular endothelium. They have primary roles in many pathophysiologic states including ischemia/reperfusion (I/R) injury. The relationships between nitric oxide (NO) and ETs are still under investigation. In this study on rats we want to focus on the interaction of NO and ET especially in I/R injury. For this purpose ET-1 and PD-156252, a nonselective ET receptor blocker, were given in a mesenteric I/R model and reactive oxygen species were detected directly using chemiluminescence of the ileal tissue. ET administrations to sham and I/R groups caused significant increases in NO concentrations whereas, in terms of peroxynitrite, which is a highly reactive group of free radicals, its increasing effects were seen only in I/R groups. This suggests that in I/R where superoxide levels increase together with NO, the conversion to peroxynitrite is likely and this effect is augmented with ET administration. On the other hand PD administration decreases superoxide and thereby peroxynitrite levels and this study shows that the effect of PD 156252 is established through this mode of action. These data suggest therapeutic approaches that may be beneficial in the treatment of I/R injury. PMID- 11418021 TI - The response of muscle interstitial prostaglandin E(2)(PGE(2)), prostacyclin I(2)(PGI(2)) and thromboxane A(2)(TXA(2)) levels during incremental dynamic exercise in humans determined by in vivo microdialysis. AB - The microdialysis in vivo technique allows the isolation, purification and quantitative determination of bioactive molecules with low molecular weight (<20.000 Da) from interstitial fluid (IF) of the muscles. PGE(2)and PGI(2)are vasodilator local hormones, while the TXA(2)is a vasoconstrictor. PGI(2)and TXA(2)are unstable and convert to stable products 6-keto-PGF(1a)and TXB(2), respectively. The purpose of this study was to evaluate the response of PGE(2), PGI(2)and TXA(2)in the IF of human muscle (vastus lateralis) during dynamic exercise with a cycle ergometer. In this study two microdialysis probes were inserted with CMA-60 microdialysis catheters into the vastus lateralis muscle of the right leg of eight healthy volunteers aged 24.1+/-2.1 years, height 177.5+/ 1.5 cm and body weight 78.1+/-2.4 kg. After insertion the microdialysis probes perfused at a rate of 3.0 microl/min with Ringer acetate solution. The dialysate fluid was collected a) during the 30' rest period, b) during the 30' exercise period at 100 watts, c) during the 30' exercise period at 150 watts and d) during the 30' rest period after exercise. Our measurements (by the RIA method) showed that the levels of PGE(2)and 6-keto-PGF(1a)in the I.F. of the vastus lateralis muscle increased significantly, while there was a significant decrease in TXB(2)during exercise. The changes in the above biomolecules were increased proportionately with the strain of the subject's muscle. CONCLUSION: Dynamic exercise of the muscles produces a local increase of the vasodilators PGE(2)and PGI(2)while the vasoconstrictor TXA(2)is reduced in the IF of the muscles. This is further evidence that exercise induces propitious biochemical changes. Furthermore, the muscle production of arachidonic acid metabolites during exercise depends on the intensity of the exercise. PMID- 11418022 TI - Effects of short-term dietary administration of marginal levels of vitamin B(6)and fish oil on lipid composition and antioxidant defences in rat tissues. AB - Previous reports have shown that vitamin B(6)deficiency leads to peroxidative stress in rat organs. In this paper, we evaluated the effects on lipid peroxidation of short-term (six weeks) dietary administration of marginal contents of vitamin B(6). A further risk factor of susceptibility to peroxidation was the presence of fish oil with higher contents of n-3 polyunsaturated fatty acid (LCPUFA). The contemporaneous vitamin B(6)deficiency and presence of fish oil caused a C18:2 increase, a C20:4 decrease, and replacement of some n-6 LCPUFA with n-3 LCPUFA, without changes in the unsaturation index. In liver, TBARS production did not show any differences between dietary conditions, whereas the activities of glutathione-dependent enzymes were stimulated. In heart, fish oil increased lipid peroxidation, especially in the vitamin B(6)-deficient group. PMID- 11418023 TI - Effect of arachidonic acid-rich oil on lipids and arachidonate metabolites in ethanol- treated rats. AB - The effects of dietary arachidonic acid-rich oil (AAoil) on lipids and arachidonate metabolites in the liver and plasma were evaluated in ethanol treated rats. Rats were fed a purified diet containing 10% weight of lard or AAoil for 14 days. Ethanol was administered by gavage at a single daily dose of 3 g/kg body weight. Comparing with the lard group, a decrease was observed in liver fatty vacuoles in the AAoil group. Plasma 6-keto-prostaglandin (PG) F1 alpha and thromboxane (TX) B(2)levels and the 6-keto-PGF1 alpha/TXB(2)ratio increased significantly in the AAoil group. Liver 6-keto-PGF1 alpha also increased but not leukotriene B(4)in the AAoil group. In the phospholipid fraction of liver tissue, plasma and red blood cells, arachidonic acid (20:4n-6) and docosatetraenoic acid (22:4n-6) increased and oleic acid (18:1n-9) and linoleic acid (18:2n-6) decreased significantly in the AAoil group compared with the lard group. These observations suggest that AAoil supplementation reduces liver injury of ethanol treated rats, although longer observation will be necessary for confirmation. PMID- 11418024 TI - Dietary n-3 and n-6 polyunsaturated oils and airway contractility. AB - Recent reports suggest modulation of chronic obstructive pulmonary disease by dietary polyunsaturated fatty acids. In the present study, we re-examined this possibility by using an established animal model of pulmonary sensitisation. Adult guinea pigs were fed diets supplemented (10% w/w) with either olive, canola or safflower oil for 4 weeks before sensitising with ovalbumin and continuing on various diets for a further 6 week period. Neither the contraction following ovalbumin challenge, nor the responses to histamine, carbachol and various eicosanoid mediators - prostaglandin F(2 alpha), leukotriene C(4), thromboxane mimetic U44619 - of isolated segments of airway tissue were altered (P>0.05, ANOVA) by the dietary lipid treatment. Lipid analysis showed changes in membrane linoleic acid (18:2n-6) and alpha -linolenic acids (alpha 18:3n-3) in lung phospholipids consistent with dietary intakes. However, no significant further desaturation/elongation of these dietary precursors was evident. Ovalbumin induced contraction was fully reversed by the lipoxygenase inhibitor esculetin whilst indomethacin resulted in a slight increase possibly due to the inhibition of bronchodilator prostanoids. Results confirm that under the conditions employed airway function was not influenced by the variable dietary intakes of n-3 and n-6 PUFA. PMID- 11418025 TI - Steady-state and time-resolved studies on oxetane cycloreversion using (thia)pyrylium salts as electron-transfer photosensitizers. AB - [reaction: see text] Cycloreversion of oxetane 1 is achieved using (thia)pyrylium salts as photosensitizers. RadicaI cation intermediates involved in the electron transfer process have been detected using laser flash photolysis. The experimental results are consistent with the reaction taking place from the triplet excited state of the sensitizer. PMID- 11418026 TI - Asymmetric synthesis of an (R)-cyanohydrin using enzymes entrapped in lens-shaped gels. AB - [structure: see text] A novel synthesis of (R)-cyanohydrins is described which is based on the use of cross-linked and subsequently poly(vinyl alcohol)-entrapped (R)-oxynitrilases. These immobilized lens-shaped biocatalysts have a well-defined macroscopic size in the mm range, show no catalyst leaching, and can be recycled efficiently. Furthermore, this immobilization method is cheap and the entrapped (R)-oxynitrilases gave similar good results compared with those of free enzymes. The (R)-cyanohydrin was obtained in good yields and with high enantioselectivities of up to >99% ee. PMID- 11418027 TI - First asymmetric synthesis of quinoline derivatives by inverse electron demand (IED) Diels-Alder reaction using chiral Ti(IV) complex. AB - [reaction: see text] (R,R)-3-Aza-3-benzyl-1,5-dihydroxy-1,5-diphenylpentane (1) ligated Ti(IV) complex (1-TiCl(2)) is used as a chiral Lewis acid catalyst for promoting asymmetric IED Diels-Alder reaction between electron-rich dienophiles and electron-poor dienes. Here we introduce a facile route for the synthesis of asymmetric tetrahydroquinoline derivatives using the above-mentioned chiral catalyst reagent in the presence of 4 A molecular sieves. The reactions proceed with moderate yields and at times high enantioselectivty. PMID- 11418028 TI - Effect of the universal base 3-nitropyrrole on the selectivity of neighboring natural bases. AB - [structure: see text] The effect of the universal base 3-nitropyrrole on the pairing selectivity of neighboring nucleosides has been determined for every combination of complementary and neighboring nucleosides. In a subset of cases the discriminatory ability of the neighboring nucleoside for its Watson-Crick complement is compromised. The results have implications for the hybridization of oligonucleotides that contain 3-nitropyrrole and suggest caution in the use of oligonucleotides that contain other universal bases. PMID- 11418029 TI - Cloning and heterologous expression of a natural product biosynthetic gene cluster from eDNA. AB - [reaction: see text] To study the natural products produced by uncultured microorganisms, an environmental DNA (eDNA) cosmid library was constructed and screened for the heterologous production of small molecules. A blue clone, CSL51, found in the eDNA library produces deoxyviolacein and the broad spectrum antibiotic violacein. The full sequence of the 6.7 kb eDNA violacein gene cluster and the characterization of violacein and deoxyviolacein from an eDNA clone are reported here. PMID- 11418030 TI - Photoinduced formation of beta-oxy-alpha,alpha-disubstituted-alpha-amino acid derivatives from ketoximes. AB - [reaction: see text] The first intermolecular radical addition onto ketoxime ethers is described. beta-Oxygenated quaternary alpha-amino acid derivatives IotaIota were obtained upon irradiation of alpha-alkoxycarbonyl ketoxime ethers Iota in the presence of suitable alpha-alkoxy carbon radical precursors and a sensitizer. PMID- 11418031 TI - Triple ring closing metathesis reaction: synthesis of adjacent cyclic ethers. AB - [reaction: see text] Adjacent tris(cyclic ethers) and enol ethers have been synthesized in good yields for the first time via a triple olefin metathesis reaction using Grubbs' catalyst RuCl(2)(=C(H)Ph)(PCy(3))(2) (Cy = cyclohexyl), and the 1,3-dimesitylimidazol-2-ylidene ruthenium benzylidene catalyst RuCl(2)(=C(H)Ph)(PCy(3))(IMes) ((IMes) = 1,3-bis(2,4,6-trimethylphenyl)imidazol-2 ylidene). The latter proved to be the most efficient catalyst in these transformations. PMID- 11418032 TI - MeOTf-mediated alkynylation of selenoamides leading to beta-methylselenenyl alpha,beta-unsaturated ketones and their characterization. AB - [reaction: see text] beta-Methylselenenyl alpha,beta-unsaturated ketones were effectively synthesized by treating selenoamides with methyl triflate, followed by reaction with lithium acetylides. The reaction proceeded with high stereoselectivity to give exclusively Z-isomers. (77)Se NMR studies and X-ray molecular structure analysis of beta-methylselenenyl alpha,beta-unsaturated ketones suggested that the products show 1,5-nonbonding interaction between the selenium atom and carbonyl oxygen atom. PMID- 11418033 TI - Palladium-catalyzed cross-coupling reaction of organoindiums with aryl halides in aqueous media. AB - [reaction: see text] Diaryl-, divinyl-, and dialkylindium proved to be stable in aqueous media and to undergo a palladium-catalyzed cross-coupling reaction with aryl halides in aqueous THF. Treatment of 3-iodophenol with diphenylindium compound, generated from indium trichloride and two equimolar amounts of a phenyl Grignard reagent, in aqueous media under palladium catalysis provided the corresponding coupling product in excellent yield. Divinyl- and diethylindium can be used for the coupling reaction in the presence of water. A wide range of functional groups, including a hydroxy group and a formyl group, are compatible with this reaction. PMID- 11418034 TI - Samarium(II)-mediated 4-exo-trig cyclization. A stereocontrolled approach to the core of pestalotiopsin A. AB - [reaction: see text] Pestalotiopsin A is a structurally unique caryophyllene-type sesquiterpene which has shown immunosuppressive activity and cytotoxicity in preliminary assays. A stereocontrolled approach to the functionalized 2 oxabicyclo[3.2.0]heptane core of pestalotiopsin A is described. The approach includes a samarium(II)-mediated 4-exo-trig cyclization and a trans-lactonization process triggered by the addition of alkylytterbium reagents to a cyclobutanone intermediate. PMID- 11418035 TI - Synthesis and characterization of oligo(9,9-dihexyl-2,7-fluorene ethynylene)s: for application as blue light-emitting diode. AB - [reaction: see text] Highly soluble and strongly blue fluorescent oligo(9,9 dihexyl-2,7-fluorene ethynylene)s (OFEs) were synthesized by a Pd/Cu-catalyzed Sonogashira coupling reaction. An organic light-emitting diode using pentamer 15 as the emitting material was successfully fabricated and emitted a bright blue light. PMID- 11418036 TI - A convergent triflate displacement approach to (alpha monofluoroalkyl)phosphonates. AB - [reaction: see text] Treatment of primary alkyl triflates or iodides with the potassium salt of diethyl (alpha-fluoro-alpha-phenylsulfonylmethyl)phosphonate yields (alpha-fluoro-alpha-phenylsulfonylalkyl)phosphonates. These can be cleanly desulfonated, in a matter of minutes, with Na(Hg) in MeOH/THF/NaH(2)PO(4). This two-step procedure complements previously reported triflate displacement approaches to alpha-nonfluorinated and alpha,alpha-difluorinated phosphonates. PMID- 11418037 TI - Palladium(II) acetate catalyzed stereoselective C-glycosidation of peracetylated glycals with arylboronic acids. AB - [reaction: see text] Addition of a variety of arylboronic acids to peracetylated glycals takes place in the presence of a catalytic amount of Pd(OAc)(2). The reaction involves the syn addition of a sigma-aryl-Pd complex to the glycal double bond followed by anti elimination of Pd(OAc)(2) to provide a carbon Ferrier type product. This method provides a practical and convenient stereoselective synthesis of C-arylglycosides. PMID- 11418038 TI - A method to accomplish a 1,4-addition reaction of bulky nucleophiles to enones and subsequent formation of reactive enolates. AB - [reaction: see text] BF(3)-promoted 1,4-addition of bulky aryl groups to alpha iodo enones, prepared from the parent enones, afforded beta-aryl-alpha-iodo ketones. Subsequent reaction with EtMgBr furnished the magnesium enolates, which upon reactions with ClP(O)(OEt)(2) and aldehydes gave enol phosphates and aldols, respectively. This method was applied successfully to a synthesis of Delta(1) trans-tetrahydrocannabinol. PMID- 11418039 TI - Enhancing reductive cleavage of aromatic carboxamides. AB - [reaction: see text] A set of aromatic and especially heteroaromatic N-benzyl carboxamides, derived from naphthalene, pyridine, pyrazine, and quinoline, and the corresponding tert-butyl acylcarbamates have been synthesized and studied by cyclic voltammetry with respect to facilitated reduction. The latter undergo regiospecific cleavage of their C(O)-N bonds under very mild reductive conditions with formation of Boc-protected (benzyl)amine in most cases in nearly quantitative yields. Examples of preparative cleavage by controlled potential electrolysis, activated aluminum, and NaBH(4) are given. PMID- 11418040 TI - Enantioselective aminolysis of an alpha-chloroester catalyzed by Candida cylindracea lipase encapsulated in sol-gel silica glass. AB - [reaction: see text] Lipase from Candida cylindracea (CcL) encapsulated in porous silica glass by a sol-gel method catalyzes enantioselective aminolysis of ethyl 2 chloropropionate. A silica matrix enhances the enzyme activity, i.e., improves the yield. The scope and limitations of the aminolysis reaction were investigated, and dynamic kinetic resolution of the ester was achieved. Encapsulated lipase remains active when used repeatedly. Encapsulation much improves the chiral discrimination by lipase and makes this enzyme even more useful in organic chemistry. PMID- 11418041 TI - The first tandem [2 + 2] cycloaddition--Michael reaction using ynolates: facile construction of substituted carbocycles. AB - [reaction: see text] A tandem [2 + 2] cycloaddition-Michael reaction using ynolate anions followed by decarboxylation produced polysubstituted five-, six-, and seven-membered cycloalkenes. PMID- 11418042 TI - Stereoselective synthesis of the axially chiral A-B ring system of vancomycin utilizing a planar chiral arene chromium complex. AB - [reaction: see text] The axial biaryl ring system of vancomycin was stereoselectively synthesized by utilizing a planar chiral tricarbonyl(arylhalide)chromium complex. Both enantiomers of the planar chiral (arylbromide)chromium complexes, (+)-9 and ent-(-)-9, can be stereoselectively transferred to an absolutely identical key intermediate 23 for the vancomycin A-B ring system by the diastereoselective Suzuki-Miyaura cross-coupling reaction as key step. PMID- 11418043 TI - Novel synthesis of alpha-amino acids via catalysis in air and water. AB - [reaction: see text] A new method was developed for the synthesis of natural and unnatural amino acid derivatives via carbon-carbon bond formation in air and water and at ambient temperature. PMID- 11418044 TI - Stereocontrolled construction of the A-ring of nitiol using a Pauson-Khand cycloaddition-ring fragmentation strategy. AB - [reaction: see text] A stereocontrolled construction of the A-ring of nitiol (1) is presented. Key features in this approach are a diastereoselective Pauson-Khand cycloaddition and a Norrish type 1 fragmentation reaction. PMID- 11418045 TI - Enamide-olefin ring-closing metathesis. AB - [reaction: see text] The first examples of ring-closing metathesis reactions of olefin-containing enamides using ruthenium-based catalysts have been demonstrated. A preliminary investigation into the scope and limitations, leading to protected five- and six-membered cyclic enamides, will be presented. PMID- 11418046 TI - Reactive ketenes through a carbonate/amine shuttle deprotonation strategy: catalytic, enantioselective alpha-bromination of acid chlorides. AB - [reaction: see text] We have developed an efficient methodology for the generation of ketenes by employing a catalytic shuttle base and potassium carbonate as a stoichiometric base. We have applied this technology to the catalytic, asymmetric alpha-bromination of acid chlorides. PMID- 11418047 TI - Asymmetric synthesis of functionalized 1,2,3,4-tetrahydroquinolines. AB - [reaction: see text] Highly enantioselective rhodium-catalyzed asymmetric hydrogenation (>98% ee) and Sharpless epoxidation (>90% ee) of o-nitrocinnamyl substrates lead to intermediates that can be transformed into tetrahydroquinoline derivatives. Starting materials are produced in high-yielding Heck reactions of an o-nitroaryl iodide and alpha-acetamidoacrylate or methyl acrylate. PMID- 11418048 TI - Design and synthesis of nonconjugated monodendrons with triarylamine repeating units. AB - [structure: see text] Design and synthesis of new monodendrons with triarylamine repeating units, which are potentially useful in charge transport applications, are described. The triarylamines are connected through a nonconjugated linkage to facilitate independent electronic tuning of the monomer units. PMID- 11418049 TI - Studies on the total synthesis of RP 66453: synthesis of fully functionalized 15 membered biaryl-containing macrocycle. AB - [structure: see text] Palladium-catalyzed Suzuki cross-coupling, Corey's enantioselective alkylation of glycine template, and macrolactamization are key steps in an efficient synthesis of the 15-membered macrocycle 2. PMID- 11418050 TI - Efficient metallo-ene reactions in organoaqueous phase. AB - [reaction: see text] The easily prepared catalyst systems PdCl(2), RhCl(2)(COD)(2), NiCl(2).6H(2)O, or Ni(COD)(2)/TPPTS have been found to form a C C bond in organoaqueous medium. Intramolecular metallo-ene reactions have been efficiently realized. Metal selectivity has been discovered. PMID- 11418051 TI - Rapid assembly of structurally defined and highly functionalized conjugated dienes via tethered enyne metathesis. AB - [reaction: see text] Conjugated dienes are versatile building blocks in organic synthesis, and the development of new methods for their synthesis remains an important topic in modern synthetic organic chemistry. We describe here an expedient synthesis of highly functionalized conjugated dienes through sequential silicon-tethered ring-closing enyne metathesis mediated by Grubbs' Ru carbene catalysts and Tamao oxidation. Notable attributes of this methodology include short synthetic manipulations and the structural complexity it confers on the resulting diene moiety. PMID- 11418052 TI - An electrochemical coupling of organic halide with aldehydes, catalytic in chromium and nickel salts. the Nozaki-Hiyama-Kishi reaction. AB - [reaction: see text] Electrochemical arylation of arenecarboxaldehydes using an iron sacrificial anode in the presence of chromium and nickel catalysts afforded the corresponding arylated secondary alcohols in moderate to good yields. The chromium and nickel salts as catalysts are obtained by oxidation of a stainless steel rod during a preelectrolysis in 7% and 3%, respectively. The process was also applied to the addition of vinyl halide, allyl acetate, or alpha-chloroester to aromatic aldehydes. PMID- 11418053 TI - Copper-catalyzed coupling of arylboronic acids and amines. AB - [reaction: see text] A general catalytic coupling of arylboronic acids and amines is reported. This room-temperature coupling was realized through the use of catalytic copper(II) acetate, 2,6-lutidine as base, and myristic acid as an additive. Functionalized aniline substrates provided the diarylamine coupling products in good yield (58-91%). A variety of alkylamines were also successfully coupled to give N-alkyl anilines in moderate yield (50-64%). PMID- 11418054 TI - Efficient syntheses of beta-cyanosugars using glycosyl iodides derived from per-O silylated mono- and disaccharides. AB - [reaction: see text] Reported herein is a general method for the efficient syntheses of a variety of beta-cyano glycosides through the activation of per-O trimethylsilyl glycosides with TMSI to form alpha-glycosyl iodides, which undergo S(N)2-type displacement when treated with tetrabutylammonium cyanide. The cyanoglycosides were reduced under mild conditions using NaBH(4) in the presence of catalytic CoCl(2)(H(2)O)(6) in THF/H(2)O to give the corresponding aminomethyl glycosides. PMID- 11418055 TI - Photoinduced group transfer radical addition of carbamotelluroates to acetylenes. AB - [reaction: see text] Te-Phenyl carbamotelluroates 1 add to acetylenes under irradiation of visible light to yield beta-telluroacrylamides 2 regioselectively. This reaction would be initiated by homolytic cleavage of the carbamoyl carbon tellurium bond, producing carbamoyl and PhTe radicals. The addition reaction proceeds via a radical chain mechanism comprising two processes: (i) addition of carbamoyl radicals at the terminal carbon of the triple bond, giving vinylic radicals, and (ii) S(H)2 reaction on the Te atom caused by the attack of the vinyl radicals to 1. PMID- 11418056 TI - Catalytic and highly efficient 1,4-addition of terminal alkynes to conjugated enones by [RuCl(2)(p-cymene)]2/pyrrolidine. AB - [reaction: see text] A wide range of terminal alkynes was added to conjugated enones in a 1,4-fashion by a ruthenium catalyst in the presence of catalytic amounts of an amine base, and the corresponding gamma,delta-alkynyl ketones were obtained in good to excellent yields. PMID- 11418058 TI - Zr-catalyzed electrophilic carbomagnesation of aryl olefins. Mechanism-based control of Zr-Mg ligand exchange. AB - [reaction: see structure] The first examples of efficient electrophilic Zr catalyzed carbomagnesations are disclosed, where in contrast to previous catalytic carbomagnesations the alkyl moiety of the electrophile is transferred (vs that of the Grignard reagent). The identity of the Grignard reagent is manipulated so that Zr-Mg exchange is facilitated, leading to the formation of alkylmagnesium halide products. PMID- 11418059 TI - Asymmetric Mukaiyama-Michael addition of acyclic enones catalyzed by allo threonine-derived B-aryloxazaborolidinones. AB - [reaction: see text] O-(2-Naphthoyl)-N-tosyl-L-allo-threonine-derived B phenyloxazaborolidinone catalyzes the asymmetric Mukaiyama-Michael addition of simple acyclic enones to give adducts of 54-85% ee. 2,6-Diisopropylphenol as an additive is demonstrated to effectively retard the undesirable Si(+)-catalyzed racemic pathway. PMID- 11418060 TI - Asymmetric synthesis of the tricyclic core of NGF-inducing cyathane diterpenes via a transition-metal-catalyzed [5 + 2] cycloaddition. AB - [reaction: see text] A concise asymmetric synthesis of the tricyclic core of cyathane diterpenes is described, based on a novel transition-metal-catalyzed intramolecular [5 + 2] cycloaddition of ynone-vinylcyclopropane 10 (assembled from commercially available (S)-(-)-limonene), which proceeds in 90% yield with >95% selectivity. This strategy provides efficient access (14 steps and 13% overall yield) to potential analogues as well as precursors of nerve growth factor (NGF)-inducing diterpenes. PMID- 11418061 TI - Construction of azaspirocyclic ketones through alpha-hydroxyiminium ion or alpha siloxy epoxide semipinacol rearrangements. AB - [reaction: see text] Semipinacol-type rearrangements to produce azaspirocyclic ketones are presented. The yields and stereoselectivities of these reactions range from 67-94% yield and 2.8:1 to 1:0 diastereoselectivity, respectively. PMID- 11418062 TI - Enantioselective formal total synthesis of (-)-dysidiolide. AB - [reaction: see text] An enantioselective formal total synthesis of the sesterterpene (-)-dysidiolide 1 beginning with an intermolecular Diels-Alder reaction of the allene ester 3 and the silyloxydiene 10 is reported. PMID- 11418063 TI - Ruthenium(II)-catalyzed cycloaddition of 1,6-diynes with isocyanates leading to bicyclic pyridones. AB - [reaction: see text] A ruthenium(II) complex possessing a planar Cp ligand, CpRu(cod)Cl, effectively catalyzed the [2 + 2 + 2] cycloaddition of 1,6-diynes with isocyanates to afford bicyclic pyridones in 58-87% yields. PMID- 11418064 TI - Allylic lithium oxyanionic directed and facilitated simmons-smith cyclopropanation: stereoselective synthesis of (+/-)-cis-sabinene hydrate and a novel ring expansion. AB - [reaction: see text] The lithium salts of acid-sensitive allyl alcohols, which themselves decompose during Simmons-Smith cyclopropanation, undergo smooth cyclopropanation in the usual stereocontrolled manner. This concept is applied to the most efficient synthesis of (+/-)-cis-sabinene hydrate and to the cyclopropanation of the anion of a nonisolable allyl alcohol resulting upon workup in a ring-expanded enone. The cyclopropanations are also faster for the lithium salts than for the allyl alcohols themselves. PMID- 11418065 TI - C(2)-symmetric Cu(II) complexes as chiral lewis acids. Catalytic, enantioselective cycloadditions of silyl ketenes. AB - [reaction: see text] C(2)-Symmetric bis(oxazoline)-Cu(II) complexes (4a-g) catalyze the enantioselective [2 + 2] cycloaddition between (silyl)ketenes and chelating carbonyl substrates. A range of substituted beta-lactones can be produced in excellent yields and selectivities. It was also found that (trimethylsilyl)ketene (1) may also undergo a highly selective hetero Diels-Alder reaction with beta,gamma-unsaturated alpha-keto esters. PMID- 11418066 TI - Synthesis and crystal structure of a tubular sexithiophene. AB - [structure: see text] The synthesis of a rigid, tubular sexithiophene is reported. Close intermolecular Cl.Cl interactions play a significant role in defining the crystal structure. PMID- 11418067 TI - Frameshift mutation in PRKDC, the gene for DNA-PKcs, in the DNA repair-defective, human, glioma-derived cell line M059J. AB - Anderson, C. W., Dunn, J. J., Freimuth, P. I., Galloway, A. M. and Allalunis Turner, M. J. Frameshift Mutation in PRKDC, the Gene for DNA-PKcs, in the DNA Repair-Defective, Human, Glioma-Derived Cell Line M059J. Radiat. Res. 156, 2-9 (2001). The glioma-derived cell line M059J is hypersensitive to ionizing radiation, lacks DNA-PK activity, and fails to express protein for the catalytic subunit, DNA-PKcs, while a sister cell line, M059K, derived from the same tumor, has normal DNA-PK activity. Both cell lines are near pentaploid and have multiple copies of chromosome 8, the chromosome on which the DNA-PKcs gene, PRKDC, is located. Sequence analysis of PCR-amplified exons revealed the loss in M059J cells of a single "A" nucleotide in exon 32, corresponding to the first nucleotide of codon 1351 (ACC, Thr) of PRKDC. Loss of the "A" nucleotide would terminate the DNA-PKcs reading frame early in exon 33. DNA from M059K cells had only the wild-type sequence. An analysis of sequences surrounding PRKDC exon 32 from 87 unrelated individuals revealed no polymorphic nucleotides except for a triplet repeat near the 3' end of this exon; no individual had a frameshift mutation in exon 32. No other sequence differences in PRKDC between M059J and M059K cells were observed in approximately 15,000 bp of genomic sequence including the sequences of exons 5 through 38 and surrounding intron sequence, suggesting a possible reduction to homozygosity at this locus prior to acquisition of the mutation leading to the M059J cell line. PMID- 11418068 TI - Gamma-ray mutagenesis studies in a new human-hamster hybrid, A(L)CD59(+/-), which has two human chromosomes 11 but is hemizygous for the CD59 gene. AB - Kraemer, S. M., Vannais, D. B., Kronenberg, A., Ueno, A. and Waldren, C. A. Gamma Ray Mutagenesis Studies in a New Human-Hamster Hybrid, A(L)CD59(+/-), which has Two Human Chromosomes 11 but is Hemizygous for the CD59 Gene. Radiat. Res. 156, 10-19 (2001). We have developed a human-CHO hybrid cell line, named A(L)CD59(+/ ), which has two copies of human chromosome 11 but is hemizygous for the CD59 gene and the CD59 cell surface antigen that it encodes. Our previous studies used the A(L) and A(L)C hybrids that respectively contain one or two sets of CHO chromosomes plus a single copy of human chromosome 11. The CD59 gene at 11p13.5 and the CD59 antigen encoded by it are the principal markers used in our mutagenesis studies. The hybrid A(L)CD59(+/-) contains two copies of human chromosome 11, only one of which carries the CD59 gene. The incidence of CD59 (-) mutants (formerly called S1(-)) induced by (137)Cs gamma rays is about fivefold greater in A(L)CD59(+/-) cells than in A(L) cells. Evidence is presented that this increase in mutant yield is due to the increased induction of certain classes of large chromosomal mutations that are lethal to A(L) cells but are tolerated in the A(L)CD59(+/-) hybrid. In addition, significantly more of the CD59 (-) mutants induced by (137)Cs gamma rays in A(L)CD59(+/-) cells display chromosomal instability than in A(L) cells. On the other hand, the yield of gamma ray-induced CD59 (-) mutants in A(L)CD59(+/-) cells is half that of the A(L)C hybrid, which also tolerates very large mutations but has only one copy of human chromosome 11. We interpret the difference in mutability as evidence that repair processes involving the homologous chromosomes 11 play a role in determining mutant yields. The A(L)CD59(+/-) hybrid provides a useful new tool for quantifying mutagenesis and shedding light on mechanisms of genetic instability and mutagenesis. PMID- 11418069 TI - Increased glomerular Vwf after kidney irradiation is not due to increased biosynthesis or endothelial cell proliferation. AB - Kuin, A., Citarella, F., Oussoren, Y. G., Van der Wal, A. F., Dewit, L. G. H. and Stewart, F. A. Increased Glomerular Vwf after Kidney Irradiation is not due to Increased Biosynthesis or Endothelial Cell Proliferation. Radiat. Res. 156, 20-27 (2001). Irradiation of the kidney induces dose-dependent, progressive renal functional impairment, which is partly mediated by vascular damage. It has previously been demonstrated that reduced renal function is preceded by an increased amount of von Willebrand factor (Vwf) in the glomerulus. The underlying mechanism and significance of this observation are unknown but, since it is an important mediator of platelet adhesion, Vwf in increased amounts could be implicated in glomerular thrombosis, resulting in impairment of renal function. Increased Vwf could be the result of increased biosynthesis by endothelial cells, or from increased numbers of endothelial cells after compensatory proliferation induced by irradiation, or it could be secondary to other events. In the present study, expression levels of mRNA for glomerular Vwf and glomerular cell proliferation rates were measured in control mouse kidneys and after irradiation with a single dose of 16 Gy. There were no significant changes in mRNA ratios for Vwf/beta-actin at 10 to 30 weeks after irradiation compared with unirradiated samples, whereas increased amounts of Vwf protein were seen in the glomeruli at these times. Labeling studies with IdU or staining for Ki67 demonstrated that glomerular proliferation was increased from 10 to 30 weeks after irradiation. Despite the increased proliferation rates, there was an absence of glomerular hyperplasia and no increase in the endothelial cell surface coverage in the glomeruli. Staining with antibodies against smooth muscle actin (SMAalpha) revealed that the observed proliferation mainly involved mesangial cells. These results indicate that the increased presence of glomerular Vwf after irradiation is not due to an increased number of endothelial cells per glomerulus, or to an increased production of Vwf. It is presumably secondary to other events, such as increased release of Vwf by damaged endothelial cells or entrapment of Vwf in the irradiated mesangial matrix. PMID- 11418070 TI - A mouse model for calculating the absorbed beta-particle dose from (131)I- and (90)Y-labeled immunoconjugates, including a method for dealing with heterogeneity in kidney and tumor. AB - Flynn, A. A., Green, A. J., Pedley, R. B., Boxer, G. M., Boden, R. and Begent, R. H. J. A Mouse Model for Calculating the Absorbed Beta-Particle Dose from (131)I- and (90)Y-Labeled Immunoconjugates, Including a Method for Dealing with Heterogeneity in Kidney and Tumor. Radiat. Res. 156, 28-35 (2001). Conventional internal radiation dosimetry methods assume that the beta-particle energy is absorbed uniformly and completely in the source organ and that the radioactivity is distributed uniformly in the source. However, in mice, a considerable proportion of the beta-particle energy can escape the source organ, resulting in large cross-organ doses. Furthermore, the distribution of radioactivity is generally heterogeneous in kidney and tumor. Therefore, a model was developed to account for cross-organ doses and for the effects of heterogeneity in kidney and tumor in mice for two of the most important radionuclides used in therapy, (131)I and (90)Y. Most mouse organs were modeled as single-compartment ellipsoids or cylinders, while heterogeneity in kidney and in tumor was addressed by using two compartments to represent the cortex and the medulla and viable and necrotic cells, respectively. The dimensions of these models were taken from previous studies, with the exception of kidney and tumor, which were defined using radioluminography and mosaics of high-power microscopy images. The absorbed fractions in each compartment were calculated using beta-particle point dose kernels. The self-organ dose was significantly higher for (131)I compared to (90)Y in all compartments, but a considerable amount of beta-particle energy was shown to escape the source organ for both radionuclides, with as much as 85% and 36% escaping the marrow for (90)Y and (131)I, respectively. The cortex was found to occupy a greater proportion of the total kidney volume than the medulla, and consequently the self-dose was higher in the cortex. In addition, the thickness of the viable shell in the tumor increased with tumor size, as did the self-dose fractions in both necrotic and viable areas. This dosimetry model improves dose estimates in mice and gives a conceptual basis for considering dosimetry in humans. PMID- 11418071 TI - Dietary eicosapentaenoic acid prevents systemic immunosuppression in mice induced by UVB radiation. AB - Moison, R. M. W. and Beijersbergen van Henegouwen, G. M. J. Dietary Eicosapentaenoic Acid Prevents Systemic Immunosuppression in Mice Induced by UVB Radiation. Radiat. Res. 156, 36-44 (2001). Reactive oxygen species (ROS) contribute to the immunosuppression induced by UVB radiation. Omega-3 fatty acids in fish oil, e.g. eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), can modulate immunoresponsiveness, but because of their susceptibility to ROS-induced damage, they can also challenge the epidermal antioxidant defense system. The influence of dietary supplementation with different omega-3 fatty acids on systemic immunosuppression induced in mice by UVB radiation was studied using the contact hypersensitivity response to trinitrochlorobenzene. In an attempt to study the mechanisms involved, UVB-radiation-induced changes in epidermal antioxidant status were also studied. Mice received high-fat (25% w/w) diets enriched with either oleic acid (control diet), EPA, DHA, or EPA + DHA (MaxEPA). Immunosuppression induced by UVB radiation was 53% in mice fed the oleic acid diet and 69% in mice fed the DHA diet. In contrast, immunosuppression was only 4% and 24% in mice fed the EPA and MaxEPA diets, respectively. Increased lipid peroxidation and decreased vitamin E levels (P < 0.05) were found in unirradiated mice fed the MaxEPA and DHA diets. For all diets, exposure to UVB radiation increased lipid peroxidation (P < 0.05), but levels of glutathione (P < 0.05) and vitamin C (P > 0.05) decreased only in the mice given fish oil. UVB irradiation did not influence vitamin E levels. In conclusion, dietary EPA, but not DHA, protects against UVB-radiation-induced immunosuppression in mice. The degree of protection appears to be related to the amount of EPA incorporated and the ability of the epidermis to maintain an adequate antioxidant level after irradiation. PMID- 11418072 TI - Thrombin promotes fibroblast proliferation during the early stages of experimental radiation pneumonitis. AB - Huang, L., Ogushi, F., Tani, K., Ogawa, H., Kawano, T., Endo, T., Izumi, K., Ueno, J., Nishitani, H. and Sone, S. Thrombin Promotes Fibroblast Proliferation during the Early Stages of Experimental Radiation Pneumonitis. Radiat. Res. 156, 45-52 (2001). To clarify the role of thrombin in the pathogenesis of radiation induced pneumonitis, we measured the thrombin activity and fibroblast growth inducing activity in bronchoalveolar lavage fluid obtained from the irradiated lungs of rats at 1, 2, 4, 8 and 18 weeks after irradiation. Thrombin activity was not detected in the bronchoalveolar lavage fluid from unirradiated rats, but the bronchoalveolar lavage fluid from irradiated rats showed significantly increased thrombin activity which reached a maximum at 4 weeks after treatment. Higher fibroblast growth-inducing activity was detected in the bronchoalveolar lavage fluid from irradiated rats at 4 and 18 weeks than in fluid from unirradiated rats. Bronchoalveolar lavage fluid from irradiated rats that were pretreated with the thrombin inhibitors antithrombin III and argatroban showed significantly inhibited fibroblast growth-inducing activity and thrombin activity at 4 weeks. However, these thrombin inhibitors did not inhibit fibroblast growth-inducing activity in bronchoalveolar lavage fluid from irradiated rats at 18 weeks. Purified rat thrombin similarly induced proliferation of fibroblasts derived from irradiated and unirradiated rats. These findings suggest that thrombin may play an important role as a fibroblast growth-inducing factor during the early stages of radiation pneumonitis. PMID- 11418073 TI - External-beam radiation therapy for improved dialysis access patency: feasibility and early safety. AB - Rectenwald, J. E., Pretus, H. A., Seeger, J. M., Huber, T. S., Mendenhall, N. P., Zlotecki, R. A., Palta, J. R., Li, Z. F., Hook, S. Y., Sarac, T. P., Welborn, M. B., Klingman, N. V., Abouhamze, Z. S. and Ozaki, C. K. External-Beam Radiation Therapy for Improved Dialysis Access Patency: Feasibility and Early Safety. Radiat. Res. 156, 53-60 (2001).Prosthetic dialysis access grafts fail secondary to neointimal hyperplasia at the venous anastomosis. We hypothesized that postoperative single-fraction external-beam radiation therapy to the venous anastomosis of hemodialysis grafts can be used safely in an effort to improve access patency. Dogs (n = 8) underwent placement of expanded polytetrafluoroethylene grafts from the right carotid artery to the left jugular vein. Five dogs received single-fraction external-beam photon irradiation (8 Gy) to the venous anastomosis after surgery. Controls were not irradiated. Shunt angiograms were completed 3 and 6 months postoperatively. Anastomoses, mid-graft, and the surrounding tissues were analyzed. Immunohistochemistry for smooth muscle cell alpha-actin, proliferating cellular nuclear antigen (PCNA), and apoptosis was performed. Incisions healed well, though all animals developed wound seromas. One control suffered graft thrombosis 4 months postoperatively. Angiography/histology confirmed severe neointimal hyperplasia at the venous anastomosis. The remaining seven dogs developed similar amounts of neointimal hyperplasia. PCNA studies showed no accelerated fibroproliferative response at irradiated anastomoses compared to controls. Skin incisions and soft tissues over irradiated anastomoses revealed no radiation-induced changes or increase in apoptosis. Thus we conclude that postoperative single-fraction external-beam irradiation of the venous anastomosis of a prosthetic arteriovenous graft that mimics the situation in humans is feasible and safe with regard to early wound healing. PMID- 11418074 TI - Thyroid cancer after diagnostic administration of iodine-131 in childhood. AB - Hahn, K., Schnell-Inderst, P., Grosche, B. and Holm, L-E. Thyroid Cancer after Diagnostic Administration of Iodine-131 in Childhood. Radiat. Res. 156, 61-70 (2001). To determine the carcinogenic effects of diagnostic amounts of (131)I on the juvenile thyroid gland, a multicenter retrospective cohort study was conducted on 4,973 subjects who either had been referred for diagnostic tests using uptake of (131)I (n = 2,262) or had had a diagnostic procedure on the thyroid without (131)I (n = 2,711) before the age of 18 years. Follow-up examinations were conducted after a mean period of 20 years after the first examination in 35% of the exposed subjects (n = 789) and in 41% of the nonexposed subjects (n = 1,118). Iodine-131 dosimetry of the thyroid was carried out according to ICRP Report No 53, and the median thyroid dose was 1.0 Gy. In the exposed group, two thyroid cancers were found during 16,500 person-years, compared to three cancers in the nonexposed group during 21,000 person-years. The relative risk for the exposed group was 0.86 (95% CI: 0.14-5.13). The study did not demonstrate an increased risk for thyroid cancer after administration of (131)I in childhood. PMID- 11418075 TI - Lenticular opacities in populations exposed to chronic low-dose-rate gamma radiation from radiocontaminated buildings in Taiwan. AB - Chen, W-L., Hwang, J-S., Hu, T-H., Chen, M-S. and Chang, W. P. Lenticular Opacities in Populations Exposed to Chronic Low-Dose-Rate Gamma Radiation from Radiocontaminated Buildings in Taiwan. Radiat. Res. 156, 71-77 (2001). Epidemiological and experimental studies have revealed the cataractogenic potential of large acute doses of radiation. However, studies on the dose response effect and the incidence of lenticular changes after exposure to chronic low doses of radiation have seldom been conducted. To evaluate quantitatively the lenticular changes in a population exposed to chronic low-dose-rate gamma radiation in their daily living or school environment in steel buildings contaminated with (60)Co in Taiwan, a total of 114 exposed individuals participated in a thorough ophthalmological examination in 1998. The lenticular opacities were evaluated by slit-lamp biomicroscopy after full pupil dilatation and were scored by the Lens Opacities Classification System III (LOCS III) and a modified subclinical minor lenticular focal defects system. These individuals were further divided into those less than 20 years old, those between 20 and 40 years old, and those more than 40 years old to evaluate the effects of age. The cumulative doses were assessed for each individual using the Taiwan Cumulative Dose (TCD) estimation system. A significant dose-dependent increase in the numbers of focal lens defects in those less than 20 years old was demonstrated, while less significant changes were observed in the other two age groups or by the LOCS III scoring. Results suggested that chronic low-dose-rate irradiation might induce minor lenticular changes, especially in lenses of young subjects. The delayed clinical changes in these young exposed subjects warrants further long-term follow-up. PMID- 11418076 TI - Analysis of a historical cohort of Chinese tin miners with arsenic, radon, cigarette smoke, and pipe smoke exposures using the biologically based two-stage clonal expansion model. AB - Hazelton, W. D., Luebeck, E. G., Heidenreich, W. F. and Moolgavkar, S. H. Analysis of a Historical Cohort of Chinese Tin Miners with Arsenic, Radon, Cigarette Smoke, and Pipe Smoke Exposures Using the Biologically Based Two-Stage Clonal Expansion Model. Radiat. Res. 156, 78-94 (2001).The two-stage clonal expansion model is used to analyze lung cancer mortality in a cohort of Yunnan tin miners based on individual histories with multiple exposures to arsenic, radon, cigarette smoke, and pipe smoke. Advances in methodology include the use of nested dose-response models for the parameters of the two-stage clonal expansion model, calculation of attributable risks for all exposure combinations, use of both a fixed lag and a gamma distribution to represent the time between generation of the first malignant cell and death from lung cancer, and scaling of biological parameters allowed by parameter identifiability. The cohort consists of 12,011 males working for the Yunnan Tin Corporation, with complete exposure records, who were initially surveyed in 1976 and followed through 1988. Tobacco and arsenic dominate the attributable risk for lung cancer. Of 842 lung cancer deaths, 21.4% are attributable to tobacco alone, 19.7% to a combination of tobacco and arsenic, 15.8% to arsenic alone, 11% to a combination of arsenic and radon, 9.2% to a combination of tobacco and radon, 8.7% to combination of arsenic, tobacco and radon, 5.5% to radon alone, and 8.7% to background. The models indicate that arsenic, radon and tobacco increase cell division, death and malignant conversion of initiated cells, but with significant differences in net cell proliferation rates in response to the different exposures. Smoking a bamboo water pipe or a Chinese long-stem pipe appears to confer less risk than cigarette use, given equivalent tobacco consumption. PMID- 11418077 TI - Translocation of MRE11 from the nucleus to the cytoplasm as a mechanism of radiosensitization by heat. AB - Zhu, W-G., Seno, J. D., Beck, B. D. and Dynlacht, J. R. Translocation of MRE11 from the Nucleus to the Cytoplasm as a Mechanism of Radiosensitization by Heat. Radiat. Res. 156, 95-102 (2001).Hyperthermia sensitizes mammalian cells to ionizing radiation, presumably by inhibiting the repair of radiation-induced double-strand breaks (DSBs). However, the mechanism by which heat inhibits DSB repair is unclear. The nuclear protein MRE11 is a component of a multi-protein complex involved in nonhomologous end joining (NHEJ) of radiation-induced DSBs. Using one-dimensional sodium dodecylsulfate polyacrylamide gel electrophoresis and Western blotting, we found that MRE11 is translocated from the nucleus to the cytoplasm when human U-1 melanoma or HeLa cells are heated for 15 min at 45.5 degrees C or when cells are heated after irradiation with 12 Gy of X rays. No such translocation is observed in unheated irradiated cells. The kinetics of migration of MRE11 to the cytoplasm was dependent upon whether the heated cells were irradiated, while the magnitude of redistribution of MRE11 was dependent upon post-treatment incubation time at 37 degrees C. Cytoplasmic MRE11 content reached a maximum 2-4 h after heating; the increase was not due to new protein synthesis. Partial recovery of nuclear MRE11 content was observed when heated cells or heated irradiated cells were incubated for up to 7 h at 37 degrees C after treatment. Western blotting results showing translocation of MRE11 from the nucleus to the cytoplasm after heating and irradiation were confirmed using confocal microscopy and immunofluorescence staining of fixed cells. Our data suggest that radiosensitization by heat may be caused, at least in part, by translocation of the DNA repair protein MRE11 from the nucleus to the cytoplasm. PMID- 11418078 TI - Enhancement of cell killing by induction of apoptosis after treatment with mild hyperthermia at 42 degrees C and cisplatin. AB - Ohtsubo, T., Igawa, H., Saito, T., Matsumoto, H., Park, H. J., Song, C. W., Kano, E. and Saito, H. Enhancement of Cell Killing by Induction of Apoptosis after Treatment with Mild Hyperthermia at 42 degrees C and Cisplatin. Radiat. Res. 156, 103-109 (2001). We examined the interactive effects of cisplatin (1.0 microg/ml) combined with hyperthermia on cell killing and on the induction of apoptosis in IMC-3 human maxillary carcinoma cells. The cytotoxic effects of hyperthermia on IMC-3 cells at 44 degrees C were greater than at 42 degrees C, as has been reported for many other cells. The induction of apoptosis, DNA fragmentation and poly(ADP-ribose) polymerase cleavage were greater after hyperthermia at 44 degrees C for 30 min compared with treatment at 42 degrees C for 105 min, even though both of these heat doses were isoeffective in reducing cell survival to 50%. Treatment with cisplatin at 37 degrees C for up to 120 min did not result in cytotoxicity or the induction of apoptosis. The enhancement ratio for treatment with cisplatin at 42 degrees C was greater than that at 44 degrees C. More apoptosis was induced after the treatment with cisplatin at 42 degrees C compared to treatment with cisplatin at 44 degrees C. Taking these findings together, the combination of cisplatin and hyperthermia at 42 degrees C appeared to be more effective than cisplatin with hyperthermia at 44 degrees C for the induction of apoptosis in IMC-3 cells. PMID- 11418079 TI - Radiation disrupts protein-DNA complexes through damage to the protein. The lac repressor-operator system. AB - Eon, S., Culard, F., Sy, D., Charlier, M. and Spotheim-Maurizot, M. Radiation Disrupts Protein-DNA Complexes through Damage to the Protein. The lac Repressor Operator System. Radiat. Res. 156, 110-117 (2001). Binding of a protein to its cognate DNA sequence is a key step in the regulation of gene expression. If radiation damage interferes with protein-DNA recognition, the entire regulation process may be perturbed. We have studied the effect of gamma rays on a model regulatory system, the E. coli lactose repressor-operator complex. We have observed the disruption of the complex upon irradiation in aerated solution. The complex is completely restored by the addition of nonirradiated repressor, but not by the addition of nonirradiated DNA. Thus radiation disrupts the DNA-protein complex by affecting the binding ability of the protein. This interpretation is supported by the dramatic loss of binding ability of a free irradiated repressor toward nonirradiated DNA. Interestingly, the dose necessary for the disruption of the irradiated complex is higher than that for inducing the complete loss of the binding ability of the free irradiated repressor. This may be due to the protection of key amino acids by the bound DNA. As seen from calculations of the accessibility of amino acids to radiolytic OH(.), the protection is due to both masking and conformational effects. PMID- 11418080 TI - Boronated dipeptide borotrimethylglycylphenylalanine as a potential boron carrier in boron neutron capture therapy for malignant brain tumors. AB - Takagaki, M., Ono, K., Masunaga, S-I., Kinashi, Y., Oda, Y., Miyatake, S-I., Hashimoto, N., Powell, W., Sood, A. and Spielvogel, B. F. Boronated Dipeptide Borotrimethylglycylphenylalanine as a Potential Boron Carrier in Boron Neutron Capture Therapy for Malignant Brain Tumors. Radiat. Res. 156, 118-122 (2001).A boronated dipeptide, borotrimethylglycylphenylalanine (BGPA), was synthesized as a possible boron carrier for boron neutron capture therapy (BNCT) for malignant brain tumors. In vitro, at equal concentrations of (10)B in the extracellular medium, BGPA had the same effect in BNCT as p-boronophenylalanine (BPA). Boron analysis was carried out using prompt gamma-ray spectrometry and track-etch autoradiography. The tumor:blood and tumor:normal brain (10)B concentration ratios were 8.9 +/- 2.1 and 3.0 +/- 1.2, respectively, in rats bearing intracranial C6 gliosarcomas using alpha-particle track autoradiography. The IC(50), i.e. the dose capable of inhibiting the growth of C6 gliosarcoma cells by 50% after 3 days of incubation, was 5.9 x 10(-3) M BGPA, which is similar to that of 6.4 x 10(-3) M for BPA. The amide bond of BGPA is free from enzymatic attack, since it is protected from hydrolysis by the presence of a boron atom at the alpha-carbon position of glycine. These results suggest promise for the use of this agent for BNCT of malignant brain tumors. Further preclinical studies of BGPA are warranted, since BGPA has advantages over both BPA and BSH. PMID- 11418081 TI - Power-frequency electromagnetic fields and exposure geometry. PMID- 11418082 TI - Low prevalence of liver-kidney microsomal autoantibodies of type 1 (LKM1) in hepatitis C seropositive subjects on Crete, Greece. AB - BACKGROUND: Hepatitis C is a serious problem on the Greek island of Crete, where a high prevalence of antibodies against hepatitis C (anti-HCV) has recently been reported. This article reports the findings of a study carried out in Crete, which investigated the prevalence of serum autoantibodies in patients with chronic hepatitis C. PATIENTS AND METHODS: One hundred and forty two patients (59 men and 83 women), who were found anti-HCV seropositive in two hospitals and two Primary Health Care Centres in Crete, were eligible. Sixty healthy blood donors (46 men, 14 women), which were negative to anti-HCV, were used as the control group. They were randomly selected from those attending Rethymnon Hospital. Autoantibodies were identified using the indirect immunofluorescence (IFL) technique on human epithelial cells from larynx cancer (HEp-2 cells), rat liver kidney-stomach substrate (CT3) and Chrithidia Luciliae (CL). RESULTS: Serum autoantibodies were detected in 104 HCV patients, yielding an overall prevalence of 73.2%. The most frequent autoantibodies were antinuclear antibodies (ANA), positive in 72 patients (50.7%). Anti-smooth muscle antibodies (ASMA) were detected in 33 patients (23.2%). Only one patient was positive for LKM1 autoantibodies. No autoantibodies were found in 38 patients (26.7%). Autoantibodies were also found in 5 out of the 60 examined healthy blood donors (8.3%). CONCLUSIONS: Autoantibodies, mainly ANA and ASMA are very common in HCV seropositive patients from Crete. By contrast LKM1 autoantibodies are exceptionally rare in these patients. PMID- 11418083 TI - Analysis of fluorescently labeled substance P analogs: binding, imaging and receptor activation. AB - BACKGROUND: Substance P (SP) is a peptide neurotransmitter found in central and peripheral nerves. SP is involved in the control of smooth muscle, inflammation and nociception. The amino acid sequence of SP is Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe Gly-Leu-Met-NH2. Five different forms of fluorescently labeled SP have recently been synthesized, in which Alexa 488, BODIPY Fl, fluorescein, Oregon Green 488 or tetramethylrhodamine has been covalently linked to SP at Lys3. Here, these novel analogs are characterized as to their ligand binding, receptor activation and fluorescence labeling properties. RESULTS: Competition binding studies, using radiolabeled [125I] SP, revealed that all of the labeled forms of SP, except for Alexa 488-SP, effectively competed with radiolabeled SP for binding at the rat SP receptor. With the exception of Alexa 488-SP, all of the SP analogs produced Ca++ elevations and fluorescence labeling of the SP receptor expressed in Chinese hamster ovary cells. In SP-responsive neurons, BODIPY Fl-SP and Oregon Green 488 SP were as effective as unlabeled SP in producing a reduction of the M-type K+ current. Fluorescein-SP produced variable results, while tetramethylrhodamine-SP was less potent and Alexa 488-SP was less effective on intact neurons. CONCLUSIONS: The above results show that fluorescent labeling of SP altered the biological activity and the binding properties of the parent peptide. Oregon Green 488 and BODIPY FL-SP are the most useful fluorophores for labeling SP without affecting its biological activity. Given these results, these probes can now be utilized in further investigations of the mechanisms of SPR function, including receptor localization, internalization and recycling. PMID- 11418084 TI - The interaction of asbestos and smoking in lung cancer. AB - Both cigarette smoke and inhaled asbestos fibres can cause lung cancer, but the assessment of how these agents act in combination is a matter of great difficulty. In non-smokers, the condition is so rare that, in any cohort of asbestos workers, the standardised mortality ratio (SMR, that is the ratio of the numbers of deaths observed and expected) is quite imprecise. The SMR for smokers, with which it has to be compared, is also subject to sampling error, making the interaction even more unstable. This accounts for much of the variation that has bedevilled evaluation. The debate has been concentrated on two hypotheses: additive (asbestos and cigarette smoke act independently) and multiplicative (asbestos produces an effect proportional to the effect of smoking). The very few data available until 1977 failed to fit the former and fitted the latter only poorly. They would have fitted better a hypothesis of greater synergism, but the only one proposed was too convoluted. So the multiplicative model appeared the only alternative, and was deemed 'accepted'. The ratio of lung cancer SMRs for non-smokers and smokers was generalised into the relative asbestos effect, RAE, with all the advantages of a parametric statistic (Berry et al., 1985, British Journal of Industrial Medicine 42, 12). On the multiplicative hypothesis, RAE=1, while RAE>1 indicates less synergism. The RAEs for the three most recent of the six results then available were >1; for one, P<0.005. From the six results combined, it was concluded that 'overall non-smokers have a relative risk of lung cancer due to asbestos that is 1.8 times that of smokers'. Some admitted uncertainty about the figure 1.8 was seized upon and even the thrust of the conclusion has been very largely disregarded. So too has the RAE and all its benefits. As a result, all later reviewers have been led into error, much of it serious: in particular, they have failed to appreciate how much of the variation arises from the inevitable imprecision of all RAEs. This failure led reviewers in 1994 to discard, quite without justification, those interactions which were less than multiplicative and came from cohort studies. Although case-referent studies seemed to support the multiplicative hypothesis, the information from them is essentially unreliable. Thus it cannot weaken the conclusions from the cohort studies, that the multiplicative hypothesis is untenable and that the relative risk of lung cancer from asbestos exposure is about twice as high in non-smokers as in smokers; the best estimate of RAE is 2.04, with 95% confidence interval 1.28-3.25. This finding is not only of high statistical significance but of great social and scientific importance. PMID- 11418085 TI - Definition and assessment of sampling and analytical accuracy. AB - Two independent definitions for quantifying measurement accuracy and two limiting schemes for their assessment are examined in this paper. Gauss' mean square error MSE is compared to the symmetric-range accuracy A, describing the range of measurements about a measurand. Both measures of accuracy account for systematic error (bias) and imprecision so as to quantify the closeness of estimates to the actual values being measured. Remarkably, it is found that the accuracy functions are closely equivalent for most method applications. Furthermore, details are presented on how to compute confidence limits on measurement accuracy so as to account for error in method evaluation. The confidence limits are qualitatively different in the case that the method undergoes extensive initial evaluation in comparison to a continual re-evaluation at each method application. To this end the statistical theories of tolerance as well as more familiar types of confidence intervals are applied. PMID- 11418086 TI - A novel synthetic erionite fibre radiolabelled with (57)Co. AB - The aim was to produce a radiolabelled fibre suitable for long-term carcinogenesis studies. To this end, we have successfully synthesised erionite fibres by a method adapted to incorporate (57)Co into the crystal framework. Morphologically the fibres are straight, of median length 2.5 microm, with 11% of fibres > 8 microm long, and median width 0.32 microm. These values are comparable to natural Oregon erionite. Autoradiography confirmed that the (57)Co was associated with the fibres themselves. The stability of the radiolabel in vivo was examined by instilling 1 mg of synthetic erionite into the lungs of F-344 rats. About half of the thoracic content of (57)Co was cleared in the first week, and over the following 5 weeks the remainder was cleared slowly with a half-time of 120 days. After 6 weeks the urinary excretion of (57)Co was only 0.054% of the initial lung content per day. This represented fibre dissolution plus any leaching of (57)Co from the fibres. It can be concluded that the (57)Co is bound into the erionite fibres with sufficient stability in vivo for studying their effects in relation to translocation to the pleura. PMID- 11418087 TI - Occupational noise exposure and hearing loss of workers in two plants in eastern Saudi Arabia. AB - OBJECTIVE: To determine the prevalence of hearing loss associated with occupational noise exposure and other risk factors. DESIGN: A cross-sectional study involving 269 exposed and 99 non-exposed subjects (non-industrial noise exposed subjects) randomly selected. Current noise exposure was estimated using both sound level meter and noise-dosimeter. Past noise exposure was estimated by interview questionnaire. Otoscopic examination and conventional frequency (0.25-8 kHz) audiometry were used to assess the hearing loss in each subject. RESULTS: 75% (202 subjects) from the exposed group were exposed to a daily Leq above the permissible level of 85 dB(A) and most (61%) of these did not and had never used any form of hearing protection. Hearing loss was found to be bilateral and symmetrical in both groups. Bivariate analysis showed a significant hearing loss in the exposed vs non-exposed subjects with a characteristic dip at 4 kHz. Thirty eight percent of exposed subjects had hearing impairment, which was an 8-fold higher rate than that found for non-exposed subjects. Multivariate analysis indicated exposure to noise was the primary, and age the secondary predictor of hearing loss. Odds of hearing impairment were lower for a small sub-group of exposed workers using hearing protection (N=19) in which logistic regression analysis showed the probability of workers adopting hearing protective devices increased with noise exposure, education, and awareness of noise control. Hearing loss was also greater amongst those who used headphones to listen to recorded cassettes. CONCLUSION: Gross occupational exposure to noise has been demonstrated to cause hearing loss and the authors believe that occupational hearing loss in Saudi Arabia is a widespread problem. Strategies of noise assessment and control are introduced which may help improve the work environment. PMID- 11418088 TI - Dustiness of different high-temperature insulation wools and refractory ceramic fibres. AB - Recent regulations are encouraging the replacement of older types of man-made mineral fibre by more soluble and, thus, less biopersistent compositions. In order for there to be any health benefits from this policy and to gain maximum benefit from such substitutions, the use of the new materials should not increase exposure. The work reported here was undertaken to investigate the use of new high-temperature glass insulation wools in place of refractory ceramic fibres (RCF). Airborne fibre levels occurring during the manufacture of both RCF and calcium magnesium silicate wools (CMS) were compared using measurements of genuine workplace exposure from a routine monitoring operation on the same plant. Exposures during use were compared in one customer facility where RCF and CMS blankets were used for the same task. Further comparisons were made in a laboratory test of dustiness using a "shaking box test". For some manufacturing tasks there are only a few workplace samples and there are few opportunities for genuine comparisons with both RCF and CMS in identical uses. However, both materials produced very similar exposure levels during manufacture, use and in the laboratory test. The novel magnesium silicate fibre was significantly dustier in the laboratory test. PMID- 11418089 TI - A study of workers' exposures to the inhalable and 'total' aerosol fractions in the primary nickel production industry using mannequins to simulate personal sampling. AB - This paper describes a study that was carried out at work sites in the primary nickel production industry to investigate the difference between inhalable and 'total' aerosol exposures by using the mannequin sampling method, and to compare the results with those from an earlier study where actual workers' personal exposures were assessed in the same way. Experiments were carried out at 21 work sites located in mining, milling, smelting, and refining works of two primary nickel production companies. During sampling, mannequins were used to simulate the physical presence of workers and the 'exposures' of these were obtained for strategic positions at selected work sites. The orientations of each mannequin with respect to the wind were rotated through 90 degrees every hour in order to simulate the approximate orientation-averaging corresponding to actual workers. Two samplers were placed side-by-side on each mannequin: the Institute of Occupational Medicine (IOM) inhalable aerosol sampler, and the 37-mm plastic cassette widely used as a personal sampler for 'total' aerosol. Each collected sample was analyzed to obtain both overall dust and overall nickel content. A total of 116 such sample pairs were collected. The results show that inhalable aerosol exposure levels-for both overall dust and for total nickel content-were consistently and significantly higher than the corresponding total aerosol exposure levels. Weighted least squares linear regression yielded (inhalable/'total') aerosol ratios ranging from 1.38 to 3.90 and 1.20 to 4.01, respectively, for overall dust and for total nickel content for different work sites. Comparison of these results with those from the earlier study of actual workers' personal exposures were in good agreement for most of the work sites studies. However, the actual intensities of exposure using the mannequin sampling method were consistently lower than those obtained from actual workers' personal sampling in our earlier study. The consistency of the (inhalable/'total') ratios between mannequin and actual personal sampling strongly suggests that the characteristics of the aerosol sampled by the two methods, most notably the particle size distribution, were the same. This in turn suggests that mannequin sampling can be useful in occupational hygiene for determining such properties of personal workers' exposures. It certainly provides a useful and cost effective method for determining factors at work sites in individual industry settings by which to examine the impact of changing exposure assessment from one based on 'total' aerosol to the recommended new approach based on inhalable aerosol. PMID- 11418090 TI - Biological monitoring the exposure to polycyclic aromatic hydrocarbons of coke oven workers in relation to smoking and genetic polymorphisms for GSTM1 and GSTT1. AB - Occupational exposure to polycyclic aromatic hydrocarbons (PAH) increases the risk of developing lung cancer. Human exposure is often demonstrated by increased internal levels of PAH metabolites and of markers for early biological effects, like DNA adducts and cytogenetic aberrations. OBJECTIVE: This study aimed to assess whether the current exposure to PAH of coke oven workers in a Dutch plant induced biological effects, and to determine if these effects are influenced by tobacco smoking and by genetic polymorphisms for the glutathione S-transferase genes GSTM1 and GSTT1. METHODS: Urinary 1-hydroxypyrene (1-OHpyr) levels were used to monitor the internal dose, while the internal effective dose was assessed by monitoring PAH-DNA adducts, DNA strand breaks (Comet assay), sister-chromatid exchanges (SCE) and cells with a high frequency of SCE (HFC) in lymphocytes together with micronuclei (MN) in exfoliated urothelial cells. RESULTS: Occupational exposure to PAH resulted in statistically significant increased 1 OHpyr levels (P<0.001), but it did not cause a significant induction of SCE, HFC, MN, DNA strand breaks or DNA adducts. Smoking caused a significant increase of 1 OHpyr (P<0.05), SCE (P<0.001), HFC (P<0.001) and DNA adducts (P<0.05), but not of MN or DNA strand breaks. Following correction for the smoking-related effects, no occupational induction of the effect biomarkers could be discerned. Multi-variate analysis did not show a significant influence of GSTM1 and GSTT1 polymorphisms on any biomarker. Also no significant interactions were observed between the various biomarkers. CONCLUSION: This study shows that in the examined plant, the occupational exposure to PAH does not result in measurable early biological effects PMID- 11418091 TI - Mixtures of nickel and cobalt chlorides induce synergistic cytotoxic effects: implications for inhalation exposure modeling. AB - Workers are often simultaneously exposed to two or more chemicals, yet little is known about the toxicity of most chemical mixtures. The traditional assumption, in the absence of further information, has been that the chemical components of a mixture have mutually independent effects, and the toxic response to multiple chemicals is additive. The data presented here show that mixtures of NiCl(2) and CoCl(2) induce a synergistic (that is, greater than additive) toxic response in cell culture. Immortalized alveolar epithelial type II cells were incubated for 4 h with various concentrations of either NiCl(2), CoCl(2), or NiCl(2) and CoCl(2) together, and cell viability assessed 24 h later. The LD(50) for NiCl(2) was 5.7 mM. CoCl(2), with an LD(50) of 1.1 mM, was about five times more potent than NiCl(2). Mixtures of NiCl(2) and CoCl(2) decreased cell viability synergistically. For example, a mixture of 750 microM NiCl(2) and 750 microM CoCl(2) reduced cell viability by more than three times the value predicted by the additive approach. We used concentration-response data from these studies in a mathematical model; this model describes the equivalent inhalation exposure to an aerosol composed of a mixture of chemicals with different toxicities and also accounts for synergistic responses to these chemicals. Our results along with previous studies using an animal model suggest that these synergisms should be taken into account when conducting future exposure assessments. PMID- 11418092 TI - The binding of cyanide to cytochrome d in intact cells, spheroplasts, membrane fragments and solubilized enzyme from Salmonella typhimurium. AB - This investigation focused on the kinetics of cyanide binding to oxidized and reduced cytochrome d in Salmonella typhimurium intact cells, spheroplasts, membrane fragments and solubilized enzyme, and on the effect of pH on this binding. Cyanide bound to the oxidized form of cytochrome d under all experimental conditions, inducing a trough at 649 nm in the oxidized-cyanide minus-oxidized difference absorption spectra. V(max) of cyanide binding to oxidized cytochrome d at pH 7.0 was 14.0+/-2.0 pmol/min/mg protein (prot.) in intact cells, 37.0+/-3.5 pmol/min/mg prot. in spheroplasts, 125.0+/-6.0 pmol/min/mg prot. in membrane fragments, and 538.0+/-8.5 pmol/min/mg prot. in solubilized cytochrome d. The pseudo-first order rate constants were 0.004 s(-1) for intact cells, 0.005 s(-1) for spheroplasts, 0.007 s(-1) for membrane fragments and 0.025 s(-1) for the solubilized enzyme. The V(max) value was highest at pH 7.0 for intact cells and solubilized cytochrome d and at pH 8.0 for both spheroplasts and membrane fragments. The K(s) of binding at pH 7.0 was around 4 mM in intact cells, spheroplasts and membrane fragments, but was 10.5 mM in solubilized cytochrome d. This difference between the K(s) values suggested a change in conformation, upon solubilization, leading to a decrease in the affinity of cyanide for the solubilized enzyme. The K(s) value was nearly the same at all pH investigated (pH 5-10). Cyanide was found to also bind to the reduced form of cytochrome d in membrane fragments (K(s)=18+/-3 mM, V(max)=377+/ 28 pmol/min/mg prot. at pH 7) and the solubilized enzyme (K(s)=18+/-1.2 mM, V(max)=649+/-45 pmol/min/mg prot. at pH 7) with a lower affinity of cyanide for the reduced cytochrome d than for the oxidized enzyme. Pseudo-first order rate constants were 0.025 s(-1) and 0.042 s(-1) respectively for membrane fragments and solubilized enzyme. The value of V(max) for cyanide binding to the reduced cytochrome d, whether membrane-bound or solubilized, increased slightly with pH (for pH 6-10) while the K(s) value dropped significantly with increasing pH. The pH dependence observed here might be interpretable as a possible role for conformational transition associated with energy transduction. Finally, this investigation pointed to the influence of the microenvironment of a protein within the cell on its reactivity. PMID- 11418093 TI - Inhibition of plant vacuolar H(+)-ATPase by diethylpyrocarbonate. AB - Treatment of the tonoplast H(+)-ATPase from mung bean seedlings (Vigna radiata L.) with histidine-specific modifier, diethyl pyrocarbonate (DEP), caused a marked loss of the ATP hydrolysis activity and the proton translocation in a concentration-dependent manner. The reaction order of inhibition was calculated to be 0.98, suggesting that at least one histidine residue of vacuolar H(+) ATPase was modified by DEP. The absorbance of the vacuolar H(+)-ATPase at 240 nm was progressively increased after incubation with DEP, suggesting that N carbethoxyhistidine had been formed. Hydroxylamine, which could break N carbethoxyhistidine, reversed the absorbance change and partially restored the enzymic activity. The pK(a) of modified residues of vacuolar H(+)-ATPase was kinetically determined to be 6.73, a value close to that of histidine. Thus, it is assuredly concluded that histidine residues of the vacuolar H(+)-ATPase were modified by DEP. Kinetic analysis showed that V(max) but not K(m) of vacuolar H(+)-ATPase was decreased by DEP. This result is interpreted as that the residual activity after DEP inhibition was primarily due to the unmodified enzyme molecules. Moreover, simultaneous presence of DEP and DCCD (N,N'-dicyclohexyl carbodiimide), an inhibitor modified at proteolipid subunit of vacuolar H(+) ATPase, did not induce synergistic inhibition, indicating their independent effects. The stoichiometry studies further demonstrate that only one out of four histidine residues modified was involved in the inhibition of vacuolar H(+) ATPase by DEP. Mg(2+)-ATP, the physiological substrate of vacuolar H(+)-ATPase, but not its analogs, exerted preferentially partial protection against DEP, indicating that the histidine residue involved in the inhibition of enzymatic activity may locate at/or near the active site and directly participate in the binding of the substrate. PMID- 11418094 TI - In vitro and in vivo electron transfer to the triheme cytochrome subunit bound to the photosynthetic reaction center complex in the purple bacterium Rhodovulum sulfidophilum. AB - The cytochrome subunit bound to the photosynthetic reaction center (RC) complex in Rhodovulum sulfidophilum lacks one heme-binding motif (CXXCH) out of four motifs found in other purple bacteria resulting in the absence of the most distal heme from the RC-core complex (S. Masuda et al., J. Biol. Chem. 274 (1999) 10795). Cytochrome c(2), which acts as the electron donor to the RC was purified, and its gene was cloned and sequenced. The redox midpoint potential of cytochrome c(2) was determined to be E(m)=357 mV. The photo-oxidation and re-reduction of purified cytochrome c(2) were observed in the presence of membrane preparations. Flash-induced photo-oxidation and re-reduction of the RC-bound cytochrome were also observed in intact cells. Despite the unusual nature of the RC-bound cytochrome subunit, the cyclic electron transfer system in Rdv. sulfidophilum was shown to be similar to those in other purple bacteria. PMID- 11418095 TI - Fluorescent probes for non-invasive bioenergetic studies of whole cyanobacterial cells. AB - Fluorescent DeltapH and DeltaPsi indicators have been screened for the non invasive monitoring of bioenergetic processes in whole cells of the cyanobacterium Synechocystis sp. PCC 6803. Acridine yellow and Acridine orange proved to be the best DeltapH indicators for the investigation of thylakoid and cytoplasmic membrane energization: While Acridine yellow indicated only cytosolic energization, Acridine orange showed signals from both the thylakoid lumen and the cytosol that could be separated kinetically. Both indicators were applied successfully to monitor cellular energetics, such as the interplay of linear and cyclic photosynthetic electron transport, osmotic adaptation and solute transport across the cytoplasmic membrane. In contrast, useful membrane potential indicators were more difficult to find, with Di-4-ANEPPS and Brilliant cresyl blue being the only promising candidates for further studies. Finally, Acridine yellow and Acridine orange could also be applied successfully for the thermophilic cyanobacterium Synechococcus elongatus. Different from Synechocystis sp. PCC 6803, where both respiration and ATP hydrolysis could be utilized for cytoplasmic membrane energization, proton extrusion at the cytoplasmic membrane in Synechococcus elongatus was preferentially driven by ATP hydrolysis. PMID- 11418096 TI - Heterogeneity of the mechanism of water splitting in photosystem II. AB - We measured the temperature dependence of oxygen evolution in thylakoids from tobacco using mass spectrometry and high resolution polarography. We determined the initial S-state distribution and the efficiency of the transition between these states including the probability of the O(2) yield through a fast mode. We observed discontinuous changes of the parameters at the temperatures 11 degrees C, 15 degrees C and 21 degrees C. Due to the mass spectroscopy data we think that the irregularity observed at 11 degrees C is due to conformational changes within the water catalytic site. We show that the different contributions of the slow and fast modes of oxygen evolution and of the water molecule exchange are correlated and that their behavior can be explained in terms of the H(2)O accessibility to the water splitting enzyme. PMID- 11418097 TI - Isolation of membrane protein subunits in their native state: evidence for selective binding of chlorophyll and carotenoid to the b(6) subunit of the cytochrome b(6)f complex. AB - Cytochrome (cyt) b-c complexes play a central role in electron transfer chains and are almost ubiquitous in nature. Although similar in their basic structure and function, the cyt b(6)f complex of photosynthetic membranes and its counterpart, the mitochondrial cyt bc(1) complex, show some characteristic differences which cannot be explained by the high resolution structure of the cyt bc(1) complex alone. Especially the presence of a chlorophyll molecule is a striking feature of all cyt b(6)f complex preparations described so far, imposing questions as to its structural and functional role. To allow a more detailed characterization, we here report the preparation of native subunits cyt b(6) and IV starting from a monomeric cyanobacterial cyt b(6)f complex. Spectroscopical and reversed-phase HPLC analyses of the purified cyt b(6) subunit showed that it contained not only two b-type hemes, but also one chlorophyll a molecule and a cyanobacterial carotenoid, echinenone. Evidence for selective binding of both pigments to this subunit is presented and their putative function is discussed. PMID- 11418098 TI - Two-dimensional structure of the native light-harvesting complex LH2 from Rubrivivax gelatinosus and of a truncated form. AB - The light-harvesting complex LH2 of Rubrivivax gelatinosus has an oligomeric structure built from alpha-beta heterodimers containing three bacteriochlorophylls and one carotenoid each. The alpha subunit (71 residues) presents a C-terminal hydrophobic extension (residues 51-71) which is prone to attack by an endogenous protease. This extension can also be cleaved by a mild thermolysin treatment, as demonstrated by electrophoresis and by matrix-assisted laser desorption-time of flight mass spectrometry. This cleavage does not affect the pigment binding sites as shown by absorption spectroscopy. Electron microscopy was used to investigate the structures of the native and thermolysin cleaved forms of the complexes. Two-dimensional crystals of the reconstituted complexes were examined after negative staining and cryomicroscopy. Projection maps at 10 A resolution were calculated, demonstrating the nonameric ring-like organization of alpha-beta subunits. The cleaved form presents the same structural features. We conclude that the LH2 complex is structurally homologous to the Rhodopseudomonas acidophila LH2. The hydrophobic C-terminal extension does not fold back in the membrane, but lays out on the periplasmic surface of the complex. PMID- 11418099 TI - Functional coupling of PSST and ND1 subunits in NADH:ubiquinone oxidoreductase established by photoaffinity labeling. AB - NADH:ubiquinone oxidoreductase (complex I) is the first, largest and most complicated enzyme of the mitochondrial electron transport chain. Photoaffinity labeling with the highly potent and specific inhibitor trifluoromethyldiazirinyl [(3)H]pyridaben ([(3)H]TDP) labels only the PSST and ND1 subunits of complex I in electron transport particles. PSST is labeled at a high-affinity site responsible for inhibition of enzymatic activity while ND1 is labeled at a low-affinity site not related to enzyme inhibition. In this study we found, as expected, that 13 complex I inhibitors decreased labeling at the PSST site without effect on ND1 labeling. However, there were striking exceptions where an apparent interaction was found between the PSST and ND1 subunits: preincubation with NADH increases PSST labeling and decreases ND1 labeling; the very weak complex I inhibitor 1 methyl-4-phenylpyridinium ion (MPP(+)) and the semiquinone analogue stigmatellin show the opposite effect with increased labeling at ND1 coupled to decreased labeling at PSST in a concentration- and time-dependent manner. MPP(+), stigmatellin and ubisemiquinone have similarly positioned centers of highly negative and positive electrostatic potential surfaces. Perhaps the common action of MPP(+) and stigmatellin on the functional coupling of the PSST and ND1 subunits is initiated by binding at a semiquinone binding site in complex I. PMID- 11418100 TI - Helicobacter mustelae lipid A structure differs from that of Helicobacter pylori. AB - The lipid A structure of the Gram-negative bacterium Helicobacter mustelae, a ferret gastric pathogen responsible for the onset of gastric diseases in its host, was investigated. Two variant lipid A structures were found in the same strain. One structure contained a bisphosphorylated beta-(1-->6)-linked D glucosamine backbone disaccharide with hydroxytetradecanoic acid in amide linkages. Unlike the structure described for the lipid A of the related human Helicobacter pylori gastric pathogen, which contains a C1 phosphate moiety, this lipid A presented phosphate groups at both the C1 and C4' positions, and contained no octadecanoyl fatty acid, which is present in H. pylori. The second lipid A structure had a different fatty acid composition in that 3-OH C(16) replaced most of the amide-linked 3-OH C(14). PMID- 11418101 TI - Expanding coincident signaling by PTEN through its inositol 1,3,4,5,6 pentakisphosphate 3-phosphatase activity. AB - PTEN, a tumor suppressor among the most commonly mutated proteins in human cancer, is recognized to be both a protein phosphatase and a phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P(3)) 3-phosphatase. Previous work [Maehama and Dixon, J. Biol. Chem. 273 (1998) 13375-13378] has led to a consensus that inositol phosphates are not physiologically relevant substrates for PTEN. In contrast, we demonstrate that PTEN is an active inositol 1,3,4,5,6 pentakisphosphate (Ins(1,3,4,5,6)P(5)) 3-phosphatase when expressed and purified from bacteria or HEK cells. Kinetic data indicate Ins(1,3,4,5,6)P(5) (K(m)=7.1 microM) and PtdIns(3,4,5)P(3) (K(m)=26 microM) compete for PTEN in vivo. Transient transfection of HEK cells with PTEN decreased Ins(1,3,4,5,6)P(5) levels. We discuss the physiological significance of these studies in relation to recent work showing that dephosphorylation of Ins(1,3,4,5,6)P(5) to inositol 1,4,5,6-tetrakisphosphate is a cell signaling event. PMID- 11418102 TI - Functions of the conserved anionic amino acids and those interacting with the substrate phosphate group of phosphoglucose isomerase. AB - Phosphoglucose isomerase catalyzes the isomerization between glucose 6-phosphate and fructose 6-phosphate in cytoplasm, and functions as autocrine motility factor and neuroleukin outside the cells. A phosphoglucose isomerase from Bacillus stearothermophilus (pgiA) was subjected to mutagenesis study to address the catalytic function of the conserved anionic residues and those probably interacting with the phosphate group of substrates. The results suggest that Glu290 works concertedly with His311 as a general acid-base pair to initiate the isomerization step, and Glu150 assists the base function of His311. The conserved loop structure consisting of Gly205-Gly206-Arg207 plays a critical role for the recognition of substrates. PMID- 11418103 TI - A novel phytyltransferase from Synechocystis sp. PCC 6803 involved in tocopherol biosynthesis. AB - The deduced polypeptide sequence of open reading frame slr1736 reveals homology to chlorophyll synthase and 1,4-dihydroxy-2-naphthoic acid phytyltransferase in Synechocystis sp. strain PCC 6803. In tocopherol and plastoquinone biosynthesis, a condensation reaction mechanistically similar to that of these two enzymes is performed. To analyze the function of this novel prenyltransferase, a deletion mutant of slr1736 was generated by homologous recombination. The mutant showed a markedly decreased tocopherol content, while plastoquinone levels remained unchanged. Since the aromatic precursor homogentisic acid accumulated in the mutant, the function of the enzyme was proven to be a novel tocopherol phytyltransferase. PMID- 11418104 TI - Peroxynitrite activates mitogen-activated protein kinase (MAPK) via a MEK independent pathway: a role for protein kinase C. AB - In this study we show that phosphorylation of extracellular signal-regulated kinase (ERK1/2; also known as p44/42MAPK) following peroxynitrite (ONOO(-)) exposure occurs via a MAPK kinase (MEK)-independent but PKC-dependent pathway in rat-1 fibroblasts. ONOO(-)-mediated ERK1/2 phosphorylation was not blocked by MEK inhibitors PD98059 and U0126. Furthermore, no increase in MEK phosphorylation was detected upon ONOO(-) treatment. Staurosporine was used to investigate whether protein kinase C (PKC) is involved. This was confirmed by down-regulation of PKC by phorbol-12,13-dibutyrate, which resulted in significant reduction of ERK1/2 phosphorylation by ONOO(-), implying that activation of ERK by ONOO(-) depends on activation of PKC. Indeed, PKCalpha and epsilon were activated upon ONOO(-) exposure. When cells were treated with ONOO(-) in a calcium-free buffer, no activation of PKCalpha was detected. Concomitantly, a reduction of ERK1/2 phosphorylation was observed suggesting that calcium was required for translocation of PKCalpha and ERK phosphorylation by ONOO(-). Indeed, ONOO(-) exposure resulted in increased cytosolic calcium, which depended on the presence of extracellular calcium. Finally, data using Go6976, an inhibitor of calcium dependent PKC activation, implied that ONOO(-)-mediated ERK1/2 phosphorylation depends on activation of a calcium-dependent PKC. PMID- 11418105 TI - Chlorophyll b is involved in long-wavelength spectral properties of light harvesting complexes LHC I and LHC II. AB - Chlorophyll (Chl) molecules attached to plant light-harvesting complexes (LHC) differ in their spectral behavior. While most Chl a and Chl b molecules give rise to absorption bands between 645 nm and 670 nm, some special Chls absorb at wavelengths longer than 700 nm. Among the Chl a/b-antennae of higher plants these are found exclusively in LHC I. In order to assign this special spectral property to one chlorophyll species we reconstituted LHC of both photosystem I (Lhca4) and photosystem II (Lhcb1) with carotenoids and only Chl a or Chl b and analyzed the effect on pigment binding, absorption and fluorescence properties. In both LHCs the Chl-binding sites of the omitted Chl species were occupied by the other species resulting in a constant total number of Chls in these complexes. 77-K spectroscopic measurements demonstrated that omission of Chl b in refolded Lhca4 resulted in a loss of long-wavelength absorption and 730-nm fluorescence emission. In Lhcb1 with only Chl b long-wavelength emission was preserved. These results clearly demonstrate the involvement of Chl b in establishing long wavelength properties. PMID- 11418106 TI - NADPH-dependent glutathione peroxidase-like proteins (Gpx-1, Gpx-2) reduce unsaturated fatty acid hydroperoxides in Synechocystis PCC 6803. AB - Here we isolated and characterized two genes (slr1171, slr1992) designated gpx-1 and gpx-2, respectively, encoding glutathione peroxidase (GPX)-like proteins (Gpx 1, Gpx-2) from Synechocystis PCC 6803. The deduced amino acid sequences for gpx-1 and gpx-2 showed high similarity to those of GPX-like proteins from higher plants and mammalian GPXs, respectively. Surprisingly, both recombinant proteins in Escherichia coli were able to utilize NADPH, but not reduced glutathione, as an electron donor and unsaturated fatty acid hydroperoxides or alkyl hydroperoxides as an acceptor. It seems accurate to refer to Gpx-1 and Gpx-2 as NADPH-dependent GPX-like proteins that serve as a new defense system for the reduction of unsaturated fatty acid hydroperoxides. PMID- 11418107 TI - LytB, a novel gene of the 2-C-methyl-D-erythritol 4-phosphate pathway of isoprenoid biosynthesis in Escherichia coli. AB - The mevalonate-independent 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway for isoprenoid biosynthesis is essential in many eubacteria, plants, and the malaria parasite. Using genetically engineered Escherichia coli cells able to utilize exogenously provided mevalonate for isoprenoid biosynthesis by the mevalonate pathway we demonstrate that the lytB gene is involved in the trunk line of the MEP pathway. Cells deleted for the essential lytB gene were viable only if the medium was supplemented with mevalonate or the cells were complemented with an episomal copy of lytB. PMID- 11418108 TI - In situ analysis of methylglyoxal metabolism in Saccharomyces cerevisiae. AB - Methylglyoxal metabolism was studied during Saccharomyces cerevisiae grown with D glucose as the sole carbon and energy source. Using for the first time a specific assay for methylglyoxal in yeast, metabolic fluxes of its formation and D-lactate production were determined. D-Glucose consumption and ethanol production were determined during growth. Metabolic fluxes were also determined in situ, at the glycolytic triose phosphate levels and glyoxalase pathway. Maximum fluxes of ethanol production and glucose consumption correspond to maxima of methylglyoxal and D-lactate formation fluxes during growth. Methylglyoxal formation is quantitatively related to glycolysis, representing 0.3% of the total glycolytic flux in S. cerevisiae. PMID- 11418109 TI - A deeper investigation on carbohydrate cycling in Sinorhizobium meliloti. AB - Recycling of triose-phosphate and pentose-phosphate was previously reported on glucose in Sinorhizobium meliloti, a polysaccharide-synthesizing bacterium, but the metabolic basis of such processes remained unclear. In this work, we have used (13)C-labelling strategies to demonstrate that carbohydrate cycling in this organism is independent of the gluconate bypass, the alternative pathway for glucose assimilation involving its periplasmic oxidation into gluconate. Furthermore, carbohydrate cycling in S. meliloti is also observed on fructose, making the situation in this bacterium significantly different from that depicted for alginate-synthesizing species. PMID- 11418110 TI - A new caffeine biosynthetic pathway in tea leaves: utilisation of adenosine released from the S-adenosyl-L-methionine cycle. AB - The four-step caffeine biosynthetic pathway includes three methylation steps that utilise S-adenosyl-L-methionine (SAM) as the methyl donor. In the process SAM is converted to S-adenosyl-L-homocysteine (SAH) which in turn is hydrolysed to L homocysteine and adenosine. Significant amounts of radioactivity from [methyl (14)C]methionine and [methyl-(14)C]SAM were incorporated into theobromine and caffeine in young tea leaf segments, and very high SAH hydrolase activity was found in cell-free extracts from young tea leaves. Substantial amounts of radioactivity from [adenosyl-(14)C]SAH were also recovered as theobromine and caffeine in tea leaf segments, indicating that adenosine derived from SAH is utilised for the synthesis of the purine ring of caffeine. From the profiles of activity of related enzymes in tea leaf extracts, it is proposed that the major route from SAM to caffeine is a SAM-->SAH-->adenosine-->adenine-->AMP-->IMP-->XMP ->xanthosine-->7-methylxanthosine-->7-methylxanthine-->theobromine-->caffeine pathway. In addition, direct adenosine kinase-catalysed formation of AMP from adenosine may participate as an alternative minor route. The activity of two of the three N-methyltransferase activities involved in caffeine biosynthesis and part of the activities of SAH hydrolase, adenosine nucleosidase, adenine phosphoribosyltransferase and adenosine kinase were located in tea chloroplasts. In contrast, no detectable activity of SAM synthetase was associated with the purified chloroplast fraction. This is a first demonstration that the purine skeleton of caffeine is synthesised from adenosine released from the SAM cycle. PMID- 11418111 TI - Influence of oocyte nuclei on demethylation of donor genome in cloned bovine embryos. AB - We recently demonstrated that satellite regions exhibit an aberrant DNA methylation in cloned bovine embryos. Here, we examined, using bisulfite sequencing technology, whether the inefficient demethylation of cloned donor genomes could be rescued by the presence of oocytic nuclei. Both AciI digestion and sequencing analyses showed that satellite sequence was demethylated more efficiently in cloned tetraploid blastocysts than in diploid clones. When methyl CpG density (the number of methyl-CpG sites per string) was scored, a significant decrease was observed in tetraploids (P<0.001). These results suggest that unknown mechanisms provided by oocytic nuclei could assist the demethylation of satellite sequences in tetraploid clones. PMID- 11418112 TI - Primary sequence requirements for S-acylation of beta(2)-adrenergic receptor peptides. AB - Palmitoylation is a post-translational modification that occurs on selected cysteines of many proteins. Since a high proportion of basic and hydrophobic residues is often found near the palmitoylated cysteine, the role of these residues in the selection of specific palmitoylation sites was assessed. Short peptides derived from the beta(2)-adrenergic receptor sequence, modified to present different proportions of basic, acidic and hydrophobic residues, were tested in an in vitro S-acylation assay. Basic residues proved to be essential, whereas hydrophobic residues greatly enhanced S-acylation and acidic residues inhibited it. Taken together, these results show that short peptides contain the required molecular determinants leading to selective S-acylation. Whether or not these sequence characteristics also contribute to the selectivity of palmitoylation in vivo will need to be further investigated. PMID- 11418113 TI - Model of interaction of the IL-1 receptor accessory protein IL-1RAcP with the IL 1beta/IL-1R(I) complex. AB - A preliminary model has been calculated for the activating interaction of the interleukin 1 receptor (IL-1R) accessory protein IL-1RAcP with the ligand/receptor complex IL-1beta/IL-1R(I). First, IL-1RAcP was modeled on the crystal structure of IL-1R(I) bound to IL-1beta. Then, the IL-1RAcP model was docked using specific programs to the crystal structure of the IL-1beta/IL-1R(I) complex. Two types of models were predicted, with comparable probability. Experimental data obtained with the use of IL-1beta peptides and antibodies, and with mutated IL-1beta proteins, support the BACK model, in which IL-1RAcP establishes contacts with the back of IL-1R(I) wrapped around IL-1beta. PMID- 11418114 TI - Hypochlorite induces the formation of LDL(-), a potentially atherogenic low density lipoprotein subspecies. AB - Oxidation of low density lipoprotein (LDL) induced by hypochlorous acid (HOCl) leading to LDL(-), a minimally oxidized subspecies of LDL, was investigated. LDL( ) is characterized by its greater electronegativity and oxidative status, and is found in plasma in vivo. Its concentration was found to be elevated under conditions that predispose humans to atherosclerosis. We found that HOCl also converts LDL rapidly to an even more oxidized state, identified as LDL(2-), which is more electronegative than LDL(-). After milder oxidation for short durations, formation of LDL(-) takes place while less LDL(2-) is formed. Under these conditions, addition of methionine not only suppressed further oxidation of LDL but also favored the formation of LDL(-) over LDL(2-), possibly by removing chloramines at lysyl residues of LDL. The presence of lipoprotein-deficient plasma did not prevent HOCl-mediated conversion of LDL to more electronegative species. It is concluded that the HOCl-mediated conversion of LDL into more electronegative species might be physiologically relevant. PMID- 11418115 TI - Fidelity of metal insertion into hydrogenases. AB - The fidelity of metal incorporation into the active center of hydrogenase 3 from Escherichia coli was studied by analyzing the inhibition of the maturation pathway by zinc and other transition metals. Hydrogenase maturation of wild-type cells was significantly affected only by concentrations of zinc or cadmium higher than 200 microM, whereas a mutant with a lesion in the nickel uptake system displayed a total blockade of the proteolytic processing of the precursor form into the mature form of the large subunit after growth in the presence of 10 microM Zn(2+). The precursor could not be processed in vitro by the maturation endopeptidase even in the presence of an excess of nickel ions. Evidence is presented that zinc does not interfere with the incorporation of iron into the metal center. Precursor of the large subunit accumulated in nickel proficient cells formed a transient substrate complex with the cognate endoprotease HycI whereas that of zinc-supplemented cells did not. The results show that zinc can intrude the nickel-dependent maturation pathway only when nickel uptake is blocked. Under this condition zinc appears to be incorporated at the nickel site of the large subunit and delivers a precursor not amenable to proteolytic processing since the interaction with the endoprotease is blocked. PMID- 11418116 TI - Human leukocyte glycosylasparaginase: cell-to-cell transfer and properties in correction of aspartylglycosaminuria. AB - Aspartylglycosaminuria (AGU), a severe lysosomal storage disease, is caused by the deficiency of the lysosomal enzyme, glycosylasparaginase (GA), and accumulation of aspartylglucosamine (GlcNAc-Asn) in tissues. Here we show that human leukocyte glycosylasparaginase can correct the metabolic defect in Epstein Barr virus (EBV)-transformed AGU lymphocytes rapidly and effectively by mannose-6 phosphate receptor-mediated endocytosis or by contact-mediated cell-to-cell transfer from normal EBV-transformed lymphocytes, and that 2-7% of normal activity is sufficient to correct the GlcNAc-Asn metabolism in the cells. Cell-to cell contact is obligatory for the transfer of GA since normal transformed lymphocytes do not excrete GA into extracellular medium. The combined evidence indicates that cell-to-cell transfer of GA plays a main role in enzyme replacement therapy of AGU by normal lymphocytes. PMID- 11418117 TI - Phytosphingosine and C2-phytoceramide induce cell death and inhibit carbachol stimulated phospholipase D activation in Chinese hamster ovary cells expressing the Caenorhabditis elegans muscarinic acetylcholine receptor. AB - Sphingolipid metabolites, such as sphingosine and ceramide, are known to play important roles in cell proliferation, differentiation and apoptosis, but the physiological roles of phytosphingosine (PHS) and phytoceramide (PHC) are poorly understood. In this study we investigated the effects of PHS, C2-PHC (N acetylPHS) and C6-PHC (N-hexanoylPHS) on cell growth and intracellular signalling enzymes. Treatment of Chinese hamster ovary (CHO) cells with PHS, C2-PHC or C6 PHC resulted in cell death in a time- and dose-dependent manner. C2-PHC induced internucleosomal DNA fragmentation, whereas PHS or C6-PHC had little if any effect on DNA fragmentation under the same experimental conditions. Both PHS and C2-PHC inhibited carbachol-induced activation of phospholipase D (PLD), but not of phospholipase C (PLC), in CHO cells expressing the Caenorhabditis elegans muscarinic acetylcholine receptor (mAChR). On the other hand, no significant effect of C6-PHC on PLD or PLC was observed. Our results show that PHS and C2-PHC exert strong cytotoxic effects on CHO cells and modulate the mAChR-mediated signal transduction pathway. PMID- 11418118 TI - Characterization of two genes, Sig1 and Sig2, encoding distinct plastid sigma factors(1) in the moss Physcomitrella patens: phylogenetic relationships to plastid sigma factors in higher plants. AB - We isolated the cDNA for a sigma factor from the moss Physcomitrella patens, which possesses unusually large N-terminal extension and the conserved subdomains 1.2-4.2. Phylogenetic analyses indicated that this novel sigma factor and PpSIG1*(2), a plastid sigma factor previously identified from Physcomitrella, were classified into SigA and SigB groups, two major classes of higher plant plastid sigma factors, respectively. According to the nomenclature recently proposed, we renamed PpSIG1* into PpSIG2, and named the novel sigma factor PpSIG1. A transient expression assay using a green fluorescent protein showed that the N-terminal region of PpSIG1 acts as a chloroplast-targeting signal. Reverse transcription-PCR experiments showed that light induces the expression of the Sig1 and Sig2 genes encoding PpSIG1 and PpSIG2, respectively. Thus, PpSIG1 and PpSIG2 are likely plastid sigma factors regulating plastid gene expression in response to light signals. PMID- 11418119 TI - Differential and opposed transcriptional effects of protein fusions containing the VP16 activation domain. AB - Overexpression of strong transcriptional activators like herpes simplex virion protein 16 (VP16) may lead to non-specific inhibition of gene expression as a result of the titration of transcription factors. Here we report that a fusion between the homeoprotein Hoxa2 and the VP16 activation domain inhibits transcription from the strong promoter/enhancers of cytomegalovirus (CMV) and Rous sarcoma virus (RSV). A similar fusion involving a Hoxa2 mutant protein that is defective in DNA binding has no effect on the CMV promoter but increases, rather than inhibits, the RSV promoter activity. This suggests that depending on its ability to bind DNA, the VP16 activator can interact with different sets of cofactors, giving rise to distinct transcriptional effects. PMID- 11418120 TI - Phosphorylation of plant actin-depolymerising factor by calmodulin-like domain protein kinase. AB - The actin-depolymerising factor (ADF)/cofilin group of proteins are stimulus responsive actin-severing proteins, members of which are regulated by reversible phosphorylation. The phosphorylation site on the maize ADF, ZmADF3, is Ser-6 but the kinase responsible is unknown [Smertenko et al., Plant J. 14 (1998) 187-193]. We have partially purified the ADF kinase(s) and found it to be calcium-regulated and inhibited by N-(6-aminohexyl)-[(3)H]5-chloro-1-naphthalenesulphonamide. Immunoblotting reveals that calmodulin-like domain protein kinase(s) (CDPK) are enriched in the purified preparation and addition of anti-CDPK to in vitro phosphorylation assays results in the inhibition of ADF phosphorylation. These data strongly suggest that plant ADF is phosphorylated by CDPK(s), a class of protein kinases unique to plants and protozoa. PMID- 11418121 TI - Histone deacetylases in replicative senescence: evidence for a senescence specific form of HDAC-2. AB - To analyze mechanisms of senescence-associated gene expression, we have investigated histone deacetylases (HDACs) in human fibroblasts undergoing replicative senescence. We found that the overall acetylation pattern of histones does not vary detectably with replicative senescence. By Northern blot and Western blot, we found a significant decrease in the abundance of HDAC-1 in senescent cells. Biochemical analysis of deacetylase activities in extracts from old and young cells revealed a striking difference. While by anion exchange chromatography we found a single peak of activity in extracts from young cells, which coincided with the elution of both HDAC-1 and HDAC-2, in senescent cells a second peak of activity was found. This second peak of activity is associated with HDAC-2 but does not contain HDAC-1. These results suggest that HDAC-2 is present in at least two distinct forms, one of which is specific for senescent cells. Further biochemical characterization of the enzyme activity revealed that addition of nicotinamide adenine dinucleotide (NAD) did not detectably influence the activity of any fraction, suggesting that NAD is not an essential co-factor for the analyzed HDACs from diploid human fibroblasts. PMID- 11418122 TI - Specificity analysis and mechanism of aurone synthesis catalyzed by aureusidin synthase, a polyphenol oxidase homolog responsible for flower coloration. AB - Aureusidin synthase, which plays a key role in the yellow coloration of snapdragon flowers, is a homolog of plant polyphenol oxidase (PPO). The enzyme specifically acted on chalcones with a 4-monohydroxy or 3,4-dihydroxy B-ring to produce aurones, for whose production the oxidative cyclization of chalcones must be preceded by 3-oxygenation. However, it exhibited virtually no PPO activity toward non-chalcone phenolics. The enzyme was competitively inhibited by phenylthiourea, a specific PPO inhibitor. These results led us to propose a mechanism of aurone synthesis by aureusidin synthase on the basis of known PPO catalyzed reactions and conclude that the enzyme is a chalcone-specific PPO specialized for aurone biosynthesis. PMID- 11418123 TI - CD spectroscopy provides evidence for excitonic interactions involving red shifted chlorophyll forms in photosystem I. AB - Selective destruction of the strongly dichroic red-shifted chlorophyll form (C709 nm) in photosystem I (PSI) trimers from Spirulina, by either non-selective high intensity illumination (photobleaching) or incubation with low concentrations of Triton X-100 is accompanied by changes in the circular dichroism spectrum of the same amplitude and of opposite sign at 677 nm. The data are interpreted in terms of a dimeric chlorophyll structure with excitonic bands at these two wavelengths. Similar photobleaching experiments with PSI-200 from maize also suggest the presence of bulk antenna/red form excitonic interactions. PMID- 11418124 TI - Electrogenic proton transfer in Rhodobacter sphaeroides reaction centers: effect of coenzyme Q(10) substitution by decylubiquinone in the Q(B) binding site. AB - An electrometric technique was used to investigate the effect of coenzyme Q(10) (UQ), substitution by decylubiquinone (dQ) at the Q(B) binding site of reaction centers (UQ-RC and dQ-RC, respectively) on the electrogenic proton transfer kinetics upon Q(B) reduction in Rhodobacter sphaeroides chromatophores. Unlike dQ RC, the kinetics of the second flash-induced proton uptake in UQ-RC clearly deviated from the mono-exponential one. The activation energy (about 30 kJ/mol) and the pH profile of the kinetics in dQ-RC were similar to those in UQ-RC, with the power law approximation used in the latter case. The interpretation of the data presumed the quinone translocation between the two binding positions within the Q(B) site. It is proposed that the native isoprenyl side chain (in contrast to decyl chain) favors the equilibrium binding of neutral quinone at the redox active 'proximal' position, but causes a higher barrier for the hydroquinone movement from 'proximal' to 'distal' position. PMID- 11418125 TI - Identification of human endomucin-1 and -2 as membrane-bound O-sialoglycoproteins with anti-adhesive activity. AB - Using a signal sequence trap method and database search, we identified a series of human cDNAs encoding two structurally related type I membrane proteins of approximately 25 kDa with multiple glycosylation motifs. These genes, termed endomucin-1/-2, are expressed in several human tissues including heart, kidney, and lung. Exogenously expressed human endomucin-1/-2 proteins were modified into 80-120 kDa glycoproteins, which were susceptible to O-sialoglycoprotein endopeptidase digestion. Transient overexpression of endomucin-1/-2 reduced the number of adhesion plaques and reduced cell attachment to the substrate. This phenotype was suppressed by laminin or the protein kinase inhibitor staurosporine. Our findings suggest that human endomucin-1/-2 negatively regulate cell adhesion to the extracellular matrix. PMID- 11418126 TI - Proposed lipocalin fold for apolipoprotein M based on bioinformatics and site directed mutagenesis. AB - Apolipoprotein M (apoM) is a novel apolipoprotein that is predominantly present in high-density lipoprotein. Sensitive sequence searches, threading and comparative model building experiments revealed apoM to be structurally related to the lipocalin protein family. In a 3D model, characterized by an eight stranded anti-parallel beta-barrel, a segment including Asn135 could adopt a closed or open conformation. Using site-directed mutagenesis, we demonstrated Asn135 in wild-type apoM to be glycosylated, suggesting that the segment is solvent exposed. ApoM displays two strong acidic patches of potential functional importance, one around the N-terminus and the other next to the opening of the beta-barrel. PMID- 11418127 TI - CSN3 interacts with IKKgamma and inhibits TNF- but not IL-1-induced NF-kappaB activation. AB - The transcription factor nuclear factor kappaB (NF-kappaB) plays a pivotal role in immune and inflammatory responses. Activation of NF-kappaB requires the activity of IKK, a kinase complex that contains two catalytic subunits, IKKalpha and IKKbeta, and a regulatory subunit IKKgamma. To understand how IKK activity is regulated, we searched for IKKgamma-interacting proteins by the yeast two-hybrid system. These screenings identified CSN3, a component of the COP9 signalsome, as a protein specifically interacting with IKKgamma. Overexpression of CSN3 inhibits NF-kappaB activation triggered by tumor necrosis factor (TNF), but not interleukin-1 (IL-1). Moreover, overexpression of CSN3 also inhibits NF-kappaB activation triggered by proteins involved in TNF signaling, including TNF-R1, TRAF2, RIP, and NIK, but not by TRAF6, a protein involved in IL-1 signaling. These data suggest that CSN3 is a specific negative regulator of TNF- but not IL 1-induced NF-kappaB activation pathways. PMID- 11418128 TI - cis-Inhibition of proteasomal degradation by viral repeats: impact of length and amino acid composition. AB - The Gly-Ala repeat (GAr) of the Epstein-Barr virus nuclear antigen 1 is a cis acting inhibitor of ubiquitin-proteasome proteolysis. We have investigated the capacity of various repeats to inhibit the turnover of the proteasomal substrate IkappaBalpha. Inhibition of TNFalpha-induced degradation was achieved by insertion of octamers containing three alanines or valines, interspersed by no more then three consecutive glycines. The inhibitory activity was abolished by increasing the length of the spacer, by eliminating the spacers, or by substitution of a single hydrophobic residue with a polar or charged residue. A serine containing octamer was inactive but inhibition was partially restored by insertion of three consecutive repeats. These findings suggest a model where inhibition requires the interaction of at least three alanine residues of the GAr in a beta-strand conformation with adjacent hydrophobic binding pockets of a putative receptor. PMID- 11418129 TI - Bidirectional activity and orientation-dependent specificity of the rat aldolase C promoter in transgenic mice. AB - We previously reported that the rat aldolase C 115 bp promoter is sufficient to ensure the brain specific expression of the chloramphenicol acetyltransferase reporter gene in transgenic mice. We identify in a further reduced 84 bp promoter several putative binding sites for the transcriptional factors Sp1, USF, AP1, and AP2. Deletion or mutation of these partially overlapping binding sites results in inactivation of the cognate transgenes. Moreover, we show that the 115 bp sequence is able to direct bidirectional transcription in vivo but, surprisingly, transcriptional activity in the opposite direction is no more brain specific. PMID- 11418130 TI - Using an in vivo phagemid system to identify non-compatible loxP sequences. AB - The site-specific recombination system of bacteriophage P1 is composed of the Cre recombinase that recognizes a 34-bp loxP site. The Cre/loxP system has been extensively used to manipulate eukaryotic genomes for functional genomic investigations. The creation of additional heterologous loxP sequences potentially expands the utility of this system, but only if these loxP sequences do not recombine with one another. We have developed a stringent in vivo assay to examine the degree of recombination between all combinations of each previously published heterologous loxP sequence. As expected, homologous loxP sequences efficiently underwent Cre-mediated recombination. However, many of the heterologous loxP pairs were able to support recombination with rates varying from 5 to 100%. Some of these loxP sequences have previously been reported to be non-compatible with one another. Our study also confirmed other heterologous loxP pairs that had previously been shown to be non-compatible, as well as defined additional combinations that could be used in designing new recombination vectors. PMID- 11418131 TI - Molecular structure of the glibenclamide binding site of the beta-cell K(ATP) channel. AB - We have investigated the structure of the glibenclamide binding site of pancreatic beta-cell ATP-sensitive potassium (K(ATP)) channels. K(ATP) channels are a complex of four pore-forming Kir6.2 subunits and four sulfonylurea receptor (SUR1) subunits. SUR1 (ABCC8) belongs to the ATP binding cassette family of proteins and has two nucleotide binding domains (NBD1 and NBD2) and 17 putative transmembrane (TM) sequences. Co-expression in a baculovirus expression system of two parts of SUR1 between NBD1 and TM12 leads to restoration of glibenclamide binding activity, whereas expression of either individual N- or C-terminal part alone gave no glibenclamide binding activity, confirming a bivalent structure of the glibenclamide binding site. By using N-terminally truncated recombinant proteins we have shown that CL3 - the cytosolic loop between TM5 and TM6 - plays a key role in formation of the N-terminal component of the glibenclamide binding site. Analysis of deletion variants of the C-terminal part of SUR1 showed that CL8 - the cytosolic loop between TM15 and TM16 - is the only determinant for the C-terminal component of the glibenclamide binding site. We suggest that in SUR1 in the native K(ATP) channel close proximity of CL3 and CL8 leads to formation of the glibenclamide binding site. PMID- 11418132 TI - Leucine and its keto acid enhance the coordinated expression of genes for branched-chain amino acid catabolism in Arabidopsis under sugar starvation. AB - Branched-chain alpha-keto acid dehydrogenase (BCKDH), a multienzyme complex, plays a key role in branched-chain amino acid catabolism. However, it remains unclear whether expression of each subunit is coordinately regulated in plants, which should be important for the efficient assembly of subunits into a functional multienzyme complex. We show that the transcripts from the Arabidopsis E1alpha subunit gene accumulated in dark-adapted leaves and in sugar-starved suspension cells. These results are complementary to our previous report that the transcripts for the E1beta and E2 subunit genes accumulated in sugar-starved cells. Expression of the E1alpha gene is likely to be regulated by hexokinase mediated sugar signaling, indicating that sugar plays a regulatory role in the coordinated expression of BCKDH subunit genes. Furthermore, Leu and its metabolite alpha-ketoisocaproate have synergistic effects on the enhanced expression of BCKDH subunit genes under sugar starvation. We hence suggest that branched-chain amino acids activate their own degradation pathway in sugar starved cells through co-induction of each subunit gene of BCKDH. PMID- 11418133 TI - Differential regulation of apoptosis in AK-5 tumor cells by the proto-oncogene Bcl-2: presence of Bcl-2 dependent and independent pathways. AB - The anti-apoptotic protein Bcl-2 functions as a crucial negative regulator of apoptosis. Bcl-2 has been shown to prevent the efflux of apoptogenic factors from mitochondria to cytosol, thus inhibiting cell death. Here, we show the susceptibility of a spontaneously regressing, rat histiocytic tumor cell line, AK 5, to the apoptotic effects of diverse stimuli and the ability of Bcl-2 overexpression to block cell death. Bcl-2 overexpression selectively inhibits apoptosis induced by ceramide and serum factor from AK-5 tumor regressing animals but not actinomycin D and curcumin, whereas the pancaspase inhibitor z-Val-Ala Asp fluoromethylketone completely blocks apoptosis, irrespective of the inducer used. The ability of Bcl-2 overexpression to block cell death does not depend on its ability to prevent cytochrome c release but correlates with its ability to prevent the dissipation of mitochondrial transmembrane potential. The results demonstrate that there are inducer dependent redundant activation pathways in a single cell, which may either be Bcl-2 dependent or independent. PMID- 11418134 TI - Electrostatic effects play a central role in cold adaptation of trypsin. AB - Organisms that live in constantly cold environments have to adapt their metabolism to low temperatures, but mechanisms of enzymatic adaptation to cold environments are not fully understood. Cold active trypsin catalyses reactions more efficiently and binds ligands more strongly in comparison to warm active trypsin. We have addressed this issue by means of comparative free energy calculations studying the binding of positively charged ligands to two trypsin homologues. Stronger inhibition of the cold active trypsin by benzamidine and positively charged P1-variants of BPTI is caused by rather subtle electrostatic effects. The different affinity of benzamidine originates solely from long range interactions, while the increased binding of P1-Lys and -Arg variants of BPTI is attributed to both long and short range effects that are enhanced in the cold active trypsin compared to the warm active counterpart. Electrostatic interactions thus provide an efficient strategy for cold adaptation of trypsin. PMID- 11418135 TI - Differential regulation of cell migration and cell cycle progression by FAK complexes with Src, PI3K, Grb7 and Grb2 in focal contacts. AB - Focal adhesion kinase (FAK) is a key mediator of integrin signaling, which has been implicated in the regulation of cell migration and cell cycle progression. Using chimeric molecules that fuse the focal adhesion targeting (FAT) sequence directly to several signaling molecules, we investigated the potential role of FAK recruitments of signaling molecules to focal contacts in the regulation of cell migration and cell cycle progression. We found that fusion of FAT to Src, the p85 subunit of phosphatidylinositol 3-kinase, Grb7 and Grb2 resulted in the efficient focal adhesion targeting of these signaling molecules. We showed that expression of Src-FAT, p85-FAT, or Grb7-FAT, but not Grb2-FAT, each stimulated cell migration. Interestingly, tyrosine phosphorylation of paxillin, but not p130cas, was induced by expression of Src-FAT, suggesting a potential role of paxillin in mediating stimulation of cell migration by the chimeric molecule. In contrast, targeting of Grb2, but not Src, p85, or Grb7, to focal contacts increased cell cycle progression. Biochemical analyses correlated Erk activation by Grb2-FAT with its stimulation of cell cycle progression. Together, these results suggest that at least part of the role of FAK interaction with these signaling molecules is to recruit them to focal contacts and that distinct FAK signaling complexes are involved in the regulation of cell migration vs. cell cycle progression. PMID- 11418136 TI - Chemical shift mapping of shikimate-3-phosphate binding to the isolated N terminal domain of 5-enolpyruvylshikimate-3-phosphate synthase. AB - To facilitate evaluation of enzyme-ligand complexes in solution, we have isolated the 26-kDa N-terminal domain of 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase for analysis by NMR spectroscopy. The isolated domain is capable of binding the substrate shikimate-3-phosphate (S3P), and this letter reports the localization of the S3P binding site using chemical shift mapping. Based on the NMR data, we propose that Ser23, Arg27, Ser197, and Tyr200 are directly involved in S3P binding. We also describe changes in the observed nuclear Overhauser effects (NOEs) that are consistent with a partial conformational change in the N terminal domain upon S3P binding. PMID- 11418137 TI - Streptomyces matensis laminaripentaose hydrolase is an 'inverting' beta-1,3 glucanase. AB - The laminaripentaose-producing beta-1,3-glucanase of Streptomyces matensis is a member of the glycoside hydrolase family GH-64. We have constructed and purified a recombinant hexahistidine-tagged form of the enzyme for characterisation. The enzyme, which exists as a monomer in solution, hydrolyses beta-1,3-glucan by a mechanism leading to overall inversion of the anomeric configuration. This is the first determination of the mechanism prevailing in glycoside hydrolase family GH 64 and this is the first characterisation of an 'inverting' beta-1,3-glucanase. PMID- 11418138 TI - pRb suppresses camptothecin-induced apoptosis in human osteosarcoma Saos-2 cells by inhibiting c-Jun N-terminal kinase. AB - This paper studies the cytotoxic effect induced by the topoisomerase I inhibitor camptothecin in human osteosarcoma Saos-2 cells, which lack p53 and contain a non functional form of the product of the retinoblastoma gene, pRb. Cytotoxicity induced by camptothecin was dose- and time-dependent; the treatment with 100 nM camptothecin reduced cell viability by 50% at 32 h and by 75% at 72 h of exposure. The cytotoxic effect was caused by apoptosis, as ascertained by morphological evidence, acridine orange-ethidium bromide staining and flow cytometric analysis. Apoptosis was accompanied by both the activation of caspase 3 and the fragmentation of poly(ADP-ribose) polymerase. Treatment with camptothecin caused a threefold increase in the activity of c-Jun N-terminal kinase (JNK) and an eightfold increase in the level of phosphorylated c-Jun. The introduction of the RB gene into Saos-2 cells reduced the rate of cell growth. Moreover, stable clones of transfected cells were resistant to camptothecin. Exposure to 100 nM camptothecin for 72 h reduced the viability of transfected cells by only 10%; moreover, very modest effects were observed on the activity of JNK as well as on the level of phosphorylated c-Jun. The results reported in this paper support the conclusion that the expression of wild-type pRb in Saos-2 cells exerts an anti-apoptotic influence through the control of JNK activity. PMID- 11418139 TI - The persistent problem with pain. PMID- 11418140 TI - Unveiling of genetic basis of resistance of S aureus to antibiotics. PMID- 11418141 TI - Bednets and malaria in Africa. PMID- 11418142 TI - Value of nerve biopsy? PMID- 11418143 TI - Euthanasia and assisted suicide: what does the Dutch vote mean? PMID- 11418144 TI - NSAIDS and selective COX-2 inhibitors: competition between gastroprotection and cardioprotection. PMID- 11418145 TI - Biological mechanisms of male infertility. PMID- 11418146 TI - Whole genome sequencing of meticillin-resistant Staphylococcus aureus. AB - BACKGROUND: Staphylococcus aureus is one of the major causes of community acquired and hospital-acquired infections. It produces numerous toxins including superantigens that cause unique disease entities such as toxic-shock syndrome and staphylococcal scarlet fever, and has acquired resistance to practically all antibiotics. Whole genome analysis is a necessary step towards future development of countermeasures against this organism. METHODS: Whole genome sequences of two related S aureus strains (N315 and Mu50) were determined by shot-gun random sequencing. N315 is a meticillin-resistant S aureus (MRSA) strain isolated in 1982, and Mu50 is an MRSA strain with vancomycin resistance isolated in 1997. The open reading frames were identified by use of GAMBLER and GLIMMER programs, and annotation of each was done with a BLAST homology search, motif analysis, and protein localisation prediction. FINDINGS: The Staphylococcus genome was composed of a complex mixture of genes, many of which seem to have been acquired by lateral gene transfer. Most of the antibiotic resistance genes were carried either by plasmids or by mobile genetic elements including a unique resistance island. Three classes of new pathogenicity islands were identified in the genome: a toxic-shock-syndrome toxin island family, exotoxin islands, and enterotoxin islands. In the latter two pathogenicity islands, clusters of exotoxin and enterotoxin genes were found closely linked with other gene clusters encoding putative pathogenic factors. The analysis also identified 70 candidates for new virulence factors. INTERPRETATION: The remarkable ability of S aureus to acquire useful genes from various organisms was revealed through the observation of genome complexity and evidence of lateral gene transfer. Repeated duplication of genes encoding superantigens explains why S aureus is capable of infecting humans of diverse genetic backgrounds, eliciting severe immune reactions. Investigation of many newly identified gene products, including the 70 putative virulence factors, will greatly improve our understanding of the biology of staphylococci and the processes of infectious diseases caused by S aureus. PMID- 11418148 TI - Effect of large-scale social marketing of insecticide-treated nets on child survival in rural Tanzania. AB - BACKGROUND: Insecticide-treated nets have proven efficacy as a malaria-control tool in Africa. However, the transition from efficacy to effectiveness cannot be taken for granted. We assessed coverage and the effect on child survival of a large-scale social marketing programme for insecticide-treated nets in two rural districts of southern Tanzania with high perennial malaria transmission. METHODS: Socially marketed insecticide-treated nets were introduced step-wise over a 2 year period from May, 1997, in a population of 480000 people. Cross-sectional coverage surveys were done at baseline and after 1, 2, and 3 years. A demographic surveillance system (DSS) was set up in an area of 60000 people to record population, births, and deaths. Within the DSS area, the effect of insecticide treated nets on child survival was assessed by a case-control approach. Cases were deaths in children aged between 1 month and 4 years. Four controls for each case were chosen from the DSS database. Use of insecticide-treated nets and potential confounding factors were assessed by questionnaire. Individual effectiveness estimates from the case-control study were combined with coverage to estimate community effectiveness. FINDINGS: Insecticide-treated net coverage of infants in the DSS area rose from less than 10% at baseline to more than 50% 3 years later. Insecticide-treated nets were associated with a 27% increase in survival in children aged 1 month to 4 years (95% CI 3-45). Coverage in such children was higher in areas with longer access to the programme. The modest average coverage achieved by 1999 in the two districts (18% in children younger than 5 years) suggests that insecticide-treated nets prevented 1 in 20 child deaths at that time. INTERPRETATION: Social marketing of insecticide-treated nets has great potential for effective malaria control in rural African settings. PMID- 11418149 TI - Activity of angiotensin-converting enzyme and risk of severe hypoglycaemia in type 1 diabetes mellitus. AB - BACKGROUND: The insertion (I) allele of the angiotensin-converting-enzyme (ACE) gene occurs at increased frequency in endurance athletes. This association suggests that low ACE activity is favourable for performance in conditions with limited substrate availability. Such conditions occur in endurance athletes during competition and in diabetic patients during insulin-induced hypoglycaemia. Patients rely on preserved functional capacity to recognise hypoglycaemic episodes and avoid progression by self-treatment. We studied whether ACE activity is related to the risk of severe hypoglycaemia in type 1 diabetes. METHODS: Consecutive adult outpatients with type 1 diabetes, untreated with ACE inhibitors or angiotensin-II-receptor antagonists (n=207) reported their experience of mild and severe hypoglycaemia during the previous 1 year and 2 years. The patients were further characterised by diabetes history, degree of hypoglycaemia awareness, measurement of C-peptide, haemoglobin A(1c), and serum ACE concentrations, and determination of ACE genotype. FINDINGS: Patients with the DD genotype had a relative risk of severe hypoglycaemia in the preceding 2 years of 3.2 (95% CI 1.4-7.4) compared with those who had the II genotype. There was a significant relation between serum ACE activity and the rate of severe hypoglycaemia (relative risk per 10 U/L increment 1.4 [1.2-1.6]), corresponding to a 3.5 times higher risk for patients in the highest quartile than for those in the lowest quartile. Multiple regression analysis showed that the effect of the ACE genotype was explained by its influence on serum ACE activity and that the only other significant determinants of the risk of severe hypoglycaemia were the degree of hypoglycaemia awareness, b-cell function, and duration of diabetes of more than 20 years. INTERPRETATION: ACE activity is a clinically significant marker of the risk of severe hypoglycaemia in patients with type 1 diabetes, especially in those with impaired defence against hypoglycaemia. These findings need to be confirmed in prospective studies. PMID- 11418150 TI - Outcome in patients with eating disorders: a 5-year study. AB - BACKGROUND: Eating disorders are disabling, unpredictable, and difficult to treat. We did a prospective 5-year investigation of a representative sample of patients with eating disorders. Our aim was to identify predictors of outcome and to assess effects of available treatments. METHODS: We prospectively investigated 95 patients with anorexia nervosa, 88 with bulimia nervosa, and 37 with eating disorders not otherwise specified (EDNOS), who sought treatment in Adelaide, South Australia. We divided patients into those who had, and had not, received treatment in specialist units and reached a safe body weight. Individuals were then further classified dependent on intensity of any treatment received. We assessed clinical symptoms, body-related attitudes, and psychosocial function. FINDINGS: 216 (98%) patients were available for follow-up after 5 years. Three patients with anorexia nervosa and two with EDNOS died. 65 (74%) bulimic, 29 (78%) EDNOS, and 53 (56%) anorexic patients had no diagnosable eating disorder. A small proportion of patients in every group had poor Morgan-Russell-Hayward scores at outcome. Final outcome was predicted by extent and intensity, but not duration, of initial symptoms in patients with anorexia nervosa, and by initial body-related attitudes and impaired psychosocial functioning in bulimia patients. We were unable to predict EDNOS outcome. Treatment did not affect outcome for any group. INTERPRETATION: Deaths in the study confirm the serious nature of eating disorders. However, our results suggest that the efficacy of existing interventions is questionable. PMID- 11418151 TI - Threats from patients and their effects on medical decision making: a cross sectional, randomised trial. AB - BACKGROUND: Negative experiences are not uncommon among doctors in Norway. Our aim was to find out about the various types of negative reactions (eg, complaints, negative exposure to the media, financial claims, and notification to the police) received by physicians from patients or relatives in response to treatment, to identify their cause, and to study their effects on subsequent clinical decisions. METHODS: We posted questionnaires about negative reactions of patients to a random sample (n=1260) of Norwegian doctors. Each doctor was additionally sent five written case simulations and asked to choose one of several proposed clinical strategies. Half (630) the physicians received cases containing threats from the patient or their relatives. FINDINGS: 988 (78%) physicians returned the questionnaire, 463 (47%) of whom reported negative experiences. Such experiences were reported more frequently by men (357 [51%]) and family physicians (157 [58%]) than by other participants. Negative experiences did not affect choice of strategy for case simulations. For the first case, chest pain, 217 (44%) physicians presented with a threat chose a defensive strategy compared with 145 (30%) of those who were not (difference 14%; 95% CI 8 20). For the second case, a headache case, the corresponding numbers were 278 (57%) and 118 (25%) (32%; 26-38). Physician age, sex, specialty, or experience of negative reactions of patients did not alter the effect of threats received during our study. INTERPRETATION: Negative experiences do not affect subsequent decision making. However, doctors do comply with wishes from patients or relatives when presented with direct threats. PMID- 11418152 TI - A blind panic. PMID- 11418153 TI - Serum soluble interleukin-2 receptor concentrations as an independent prognostic marker in head and neck cancer. AB - Currently head and neck squamous cell-carcinomas are staged clinically, though this is not ideal. We did a multivariate prospective study of 234 patients with head and neck squamous-cell carcinoma and showed that high serum concentrations of sIL-2Ralpha at diagnosis were highly correlated with a shorter survival (p<0.0001). In addition, patients who had low serum sIL-2Ralpha concentrations at diagnosis were less likely to develop distant metastasis during the 36 months follow up compared with the group with high serum sIL-2Ralpha concentrations (p<0.001). These findings suggest that serum sIL-2Ralpha could be considered as an independent serum biomarker in head and neck cancer patients. PMID- 11418154 TI - Psychiatric illness and mortality after hip fracture. AB - People with dementia or delirium have increased mortality in the 6 months after hip fracture, but depression might take longer to have an effect. We assessed the psychiatric status of 731 participants with hip fracture and analysed the effect of psychiatric illness on mortality during the next 2 years. We found that dementia, delirium, and depression all increased the risk of mortality (p<0.0001, p<0.0001, and p=0.0359, respectively), and that mortality differed significantly between hospitals (p=0.0003). We suggest that psychiatric interventions should be asssessed in hip-fracture patients with adequate follow up of outcome measures. PMID- 11418155 TI - A common mitochondrial DNA variant associated with susceptibility to dilated cardiomyopathy in two different populations. AB - Idiopathic dilated cardiomyopathy is a recognised manifestation of mitochondrial disease due to specific mitochondrial (mt) DNA mutations. However, whether mtDNA polymorphisms predispose to sporadic dilated cardiomyopathy is not known. We analysed two populations with this disorder for a general mtDNA variant (T16189C), previously implicated in susceptibility to type 2 diabetes. We noted an increased frequency of the polymorphism in both populations compared with controls (p=0.002). The polymorphism occurred on different mtDNA backgrounds, suggesting that it might be a functional variant. This association of an mtDNA variant with increased susceptibility to dilated cardiomyopathy provides evidence for a mitochondrial cause in sporadic disease. PMID- 11418156 TI - Airborne outbreak of nosocomial Scedosporium prolificans infection. AB - We describe six inpatients with acute non-lymphocytic leukaemia who developed invasive infection with Scedosporium prolificans resistant to amphotericin B, flucytosine, ketoconazole, fluconazole, and itraconazole. All six patients died. Phenotypic and genotypic assessment of samples from clinical material and ambient air from the isolation rooms where the patients were being treated showed that the epidemic was caused by a single strain. After implementation of aerial control measures, there were no further infections with this organism. We conclude that fatal multidrug-resistant S prolificans epidemics can be aerially transmitted and can be prevented with implementation of appropriate infection control measures. PMID- 11418157 TI - Pakistan investigates black market sale of HIV kits. PMID- 11418158 TI - Contraceptives do not protect against pelvic inflammatory disease. PMID- 11418160 TI - Raloxifene fails to slow cognitive decline in older women. PMID- 11418161 TI - Early visual experience essential for facial recognition. PMID- 11418164 TI - Meaning of morning sickness still unsettled. PMID- 11418165 TI - Controlling phantom limb pain in Sierra Leone. PMID- 11418166 TI - UK prison doctors warn that prison health care is falling apart. PMID- 11418167 TI - Doctors in Kenya call for fair mental health policy. PMID- 11418169 TI - USA consider national programmes to oversee human research. PMID- 11418171 TI - Europe tackles consumers fears over food safety. PMID- 11418172 TI - Pacing for heart failure. AB - Pacing for patients with severe heart failure without bradyarrhythmia has been proposed as an addition to medical therapy over the past decade. Alteration of the normal electrical activation sequence of the heart modifies its mechanical action, especially when ventricular function is poor. Both the site of ventricular-lead placement and timing with the atria have been manipulated in attempts to alleviate the symptoms of heart failure. Most recently, in addition to the conventional two leads used for pacing, a third lead to pace the left ventricle has been advocated in some patients with heart failure. We review the evidence for pacing in heart failure. PMID- 11418173 TI - Endemic stability--a veterinary idea applied to human public health. AB - Endemic stability is an epidemiological state of a population, in which clinical disease is scarce despite high level of infection. The notion was developed to describe patterns of tick-borne disease in cattle. However, we propose a general model of endemic stability that is applicable to a broader range of diseases that are important in public health, including malaria, rubella, and mumps. We postulate that endemic stability requires only that (1) the probability, or severity, of clinical disease after infection increases with age, and (2) after one infection, the probability that subsequent infections result in disease is reduced. We present these criteria in simple mathematical terms. Our hypothesis predicts that partial disease control activities might, under certain circumstances, lead to an increase in disease incidence. We discuss the implications for public health interventions. PMID- 11418180 TI - Illiterature and medicine. PMID- 11418181 TI - Seeing is believing: imaging techniques to monitor plant health. AB - Historically, early stress-induced changes in plants have been mainly detected after destructive sampling followed by biochemical and molecular determinations. Imaging techniques that allow immediate detection of stress-situations, before visual symptoms appear and adverse effects become established, are emerging as promising tools for crop yield management. Such monitoring approaches can also be applied to screen plant populations for mutants with increased stress tolerance. At the laboratory scale, different imaging methods can be tested and one or a combination best suited for crop surveillance chosen. The system of choice can be applied under controlled laboratory conditions to guide selective sampling for the molecular characterisation of rapid stress-induced changes. Such an approach permits to isolate presymptomatically induced genes, or to obtain a panoramic view of early gene expression using gene-arrays when plants undergo physiological changes undetected by the human eye. Using this knowledge, plants can be engineered to be more stress resistant, and tested for field performance by the same methodologies. In ongoing efforts of genome characterisation, genes of unknown function are revealed at an ever-accelerating pace. By monitoring changes in phenotypic characteristics of transgenic plants expressing those genes, imaging techniques could help to identify their function. PMID- 11418182 TI - Nd1, a novel murine Kelch family protein, may play the role of a housekeeping gene. AB - The murine Nd1 gene encodes a novel Kelch family protein and expresses two forms of mRNA, long (Nd1-L) and short (Nd1-S), in various tissues. We characterized the genomic organization of the Nd1 gene, and found that Nd1-L and Nd1-S consist of 16 and nine exons respectively, and that exons I-VIII are shared between them. Three transcription initiation sites were identified in the 5'-flanking region and the most 3' side (+1) is likely to be a major one. Promoter analysis revealed that the region between positions -247 and -86 was sufficient for expression, and that two Sp1-binding sites and one NF-kappaB-binding site in the region were critical for promoter activity. Furthermore, the promoter region lacks a TATA and a CAAT box and has a highly GC-rich region with two important Sp1-binding sites. These characteristics of the Nd1 gene promoter are similar to the properties of housekeeping genes. PMID- 11418183 TI - Heterogeneous DNA binding modes of berenil. AB - Isothermal titration calorimetry (ITC) profiles of berenil bound to different DNAs show that, despite the strong preference of berenil for AT-rich regions in DNA, it can bind to other DNA sequences significantly. The ITC results were used to quantify the binding of berenil, and the thermodynamic profiles were obtained using natural DNAs as well as synthetic polynucleotides. ITC binding isotherms cannot be simply described when a single set of identical binding sites is considered, except for poly[d(A-T)2]. Ultraviolet melting of DNA and differential scanning calorimetry were also used to quantify several aspects of the binding of berenil to salmon testes DNA. We present evidence for secondary binding sites for berenil in DNA, corresponding to G+C rich sites. Berenil binding to poly[d(G-C)2] is also observed. Circular dichroism experiments showed that binding to GC-rich sites involves drug intercalation. Using a molecular modeling approach we demonstrate that intercalation of berenil into CpG steps is sterically feasible. PMID- 11418184 TI - Structure of the Hodgkin's lymphoma-associated human CD30 gene and the influence of a microsatellite region on its expression in CD30(+) cell lines. AB - The CD30 antigen is a member of the tumor necrosis factor receptor (TNFR) family which is overexpressed on the surface of the tumor cells of Hodgkin's lymphoma, anaplastic large cell lymphoma (ALCL), and embryonal carcinoma of the testis. In this study the entire cd30 gene which is more than 24000 bp long and organized in eight exons was characterized by analyzing cosmid and phage lambda clones from human placental libraries with long-range polymerase chain reaction (PCR) and sequencing. Differences to other genes of the TNFR family were detected in the region encoding the extracellular domain of the cd30 gene. In nearly all other TNFR genes, the coding region of each cysteine-rich repeat is interrupted by one intron, i.e., the 3-4 cysteine-rich repeats of these receptors are encoded by at least 4-5 exons, whereas the six cysteine-rich repeats of the cd30 gene are encoded by two exons, i.e., each of these exons encode three cysteine-rich repeats. In addition, we also found a genetic polymorphism of tetranucleotide ATCC-repeats in the 5' part of the CD30 promoter. This region was amplified by PCR from seven CD30 overexpressing human lymphoid cell lines and five human tissues with an absent or very low CD30 expression. The amplification products showed length differences of more than 550 bp. The number of the ATCC-repeats was higher in CD30(+) cell lines than in normal tissues. Comparison of the individual PCR products in reporter gene assays revealed that the CD30 promoter activity increased with the length of this polymorphic region up to eightfold. The data suggest that the number of ATCC-repeats in the 5' region of the CD30 promoter modulates the regulation of CD30 expression. PMID- 11418185 TI - Non-specific deadenylation and deguanylation of naked RNA catalyzed by ricin under acidic condition. AB - Ricin A-chain catalyzes the hydrolysis of the N-glycosidic bond of a conserved adenosine residue at position 4324 in the sarcin/ricin domain of 28S RNA of rat ribosome. The GAGA tetraloop closed by C-G pairs is required for recognition of the cleavage site on 28S ribosomal RNA by ricin A-chain. In this study, ricin A chain (reduced ricin) exhibits specific depurination on a synthetic oligoribonucleotide (named SRD RNA) mimic of the sarcin/ricin domain of rat 28S ribosomal RNA under neutral and weak acidic conditions. Furthermore, the activity of intact ricin is also similar to that of ricin A-chain. However, under more acidic conditions, both enzymes lose their site specificity. The alteration in specificity of depurination is not dependent on the GAGA tetraloop of SRD RNA. A higher concentration of KCl inhibits the non-specific N-glycosidase activity much more than the specific activity of ricin A-chain. In addition, characterization of depurination sites by RNA sequencing reveals that under acidic conditions ricin A-chain can release not only adenines, but also guanines from SRD RNA or 5S ribosomal RNA. This is the first report of the non-specific deadenylation and deguanylation activity of ricin A-chain to the naked RNA under acidic conditions. PMID- 11418186 TI - Wheat mitochondria ccmB encodes the membrane domain of a putative ABC transporter involved in cytochrome c biogenesis. AB - Assembly of cytochromes c is mediated by different proteins depending on the organism and organelle considered. In land plants, mitochondria follow a pathway distinct from that of yeast and animal mitochondria, more similar to that described for alpha- and gamma-proteobacteria. Indeed, in plant mitochondria, four genes were identified based on the similarities of their products with bacterial proteins involved in c-type cytochrome maturation. We report the characterisation of one of these mitochondrial genes in Triticum aestivum, TaccmB, which is proposed to encode a subunit of an ABC transporter. The transcript extremities were mapped and cDNA sequencing revealed 42 C to U editing positions in the 618 nucleotide long coding region. This high editing rate affects the identity of 32 amino acids out of 206. Antibodies directed against wheat CcmB recognise a 28 kDa protein in an enriched inner mitochondrial membrane protein fraction, a location which is in agreement with the high hydrophobicity of the protein and its function as a putative transmembrane domain of an ABC transporter involved in cytochrome c and c1 biogenesis in plant mitochondria. PMID- 11418187 TI - Cap-independent translation conferred by the 5'-untranslated region of human thymidine kinase mRNA. AB - Translational control is one of the mechanisms that regulate thymidine kinase (TK) expression in the cell cycle. Evidence for the TK mRNA sequence that is involved in its own translation has been lacking. In this report, we show that TK deficient mouse fibroblasts transfected with pFLAG-TK express a TK mRNA containing the 5'-untranslated region (5'UTR) and produce two polypeptides, FLAG TK and TK, resulting from an alternative initiation of translation. Most interestingly, the 5'UTR of TK allowed the translation of FLAG-TK mRNA to become cap-independent in an in vitro translation system. Furthermore, this 5'UTR sequence decreased significantly the efficiency of translation from the AUG codon of FLAG when the concentration of FLAG-TK RNA was low. Here, we also show that in normal human IMR-90 fibroblasts the induction of TK polypeptide by serum stimulation is insensitive to rapamycin treatment, which is known to inhibit the translations of transcripts of some growth-controlled genes by affecting the cap binding efficiency. Taken together, we propose that the 5'UTR in TK mRNA might actually confer a secondary structure to regulate ribosome binding during translation in a cap-independent manner. PMID- 11418188 TI - Genomic structure and promoter characterization of an imprinted tumor suppressor gene ARHI. AB - We have recently identified a maternally imprinted tumor suppressor gene, ARHI (aplysia ras homolog I), the expression of which is lost in ovarian and breast cancers. We have now characterized the genomic structure of the gene including its promoter and the methylation status of its upstream CpG islands. The ARHI gene spans approximately 8 kb containing two exons and one intron. Exon 1 contains 81 non-translated nucleotides, connected to exon 2 with a 3.2-kb intron. The entire protein-coding region is located within exon 2 and encodes a 229 residue small GTP-binding protein belonging to the Ras superfamily. Genomic structure analysis has identified three potential CpG islands. Two of them (CpG island I and II) are located within the promoter and adjacent exon 1 of the ARHI gene. Aberrant methylation of these CpG islands has been detected in breast cancer cells but not in normal epithelial cells, supporting the possibility that appropriate methylation status of the CpG islands in the promoter region may play a role in the downregulation of ARHI gene expression. A TATA box is found 27 bp upstream of the transcription start site associated with several putative transcription factor binding sites. Transient transfection with nested deletion constructs of the 2-kb ARHI promoter regions fused to a luciferase reporter indicated a 121-bp sequence upstream of the transcription initiation site is required for basal promoter activity. Interestingly, this is the region where lower promoter activity has been observed in cancer cells than in normal cells. PMID- 11418189 TI - Cloning and characterization of the functional promoter of mouse estrogen receptor beta gene. AB - The recently discovered estrogen receptor beta (ERbeta) exhibits some properties distinct from those of the classical estrogen receptor, ERalpha. To elucidate the mechanism underlying the regulation of ERbeta gene expression, we cloned and characterized the 5'-flanking (promoter) region of the mouse ERbeta (mERbeta) gene. A TATA-like motif was found in the 5'-flanking region, and transcription initiation sites were mapped 24 bp and 27 bp downstream from the motif by primer extension analysis and 5'-rapid amplification of cDNA ends. The mERbeta promoter contains several putative cis-acting elements, many of which are also found in the mouse ERalpha promoter. Therefore, the expression of both ERs may be partially regulated by a common mechanism. Luciferase assays revealed that the mERbeta promoter activity paralleled the endogenous expression levels of mERbeta in several cell lines. Successive 5'-deletion analyses showed that element(s) critical for its basal activity and negative regulatory element(s) are located in the sequences -90/+33 and -505/-372, respectively. The characterization of the mERbeta promoter will allow further studies to investigate the transcriptional regulation of mERbeta. PMID- 11418190 TI - Differences in DNA-sequence recognition between the DNA-binding domain fragment and the full-length molecule of the heat-shock transcription factor of schistosome. AB - Binding and inhibition studies reveal that the DNA-binding domain (DBD) fragment and the full-length molecule of the heat-shock transcription factor of schistosome (SmHSF) differ in DNA sequence recognition. SmHSF does not recognize the ideal HSE consensus sequence (nGAAnnTTCnnGAAn) but recognizes a variant HSE that contains nGTAn instead of nGAAn in the third pentamer. The DBD reacts efficiently with the ideal HSE sequence and with lower affinity with the variant HSE sequence. These findings suggest that elements inside and outside the DBD contribute to the DNA-binding specificity of HSF. PMID- 11418191 TI - mRNA sequence of the Xenopus laevis paxillin gene and its expression. AB - Paxillin is a cytoskeletal protein found in structures of focal adhesions where cells adhere to the extracellular matrix. We isolated paxillin cDNA from the Xenopus laevis ovary. The cDNA sequence encodes a protein of 539 amino acids with four LIM and five LD motifs. 80% of the amino acids of frog paxillin are shared by human and chicken paxillins. Northern analysis showed that the frog gene is expressed in the spleen, kidney, testis and ovary. Immunocytochemistry showed that paxillin protein is accumulated in the nucleus as well as in the periphery of the cytoplasm of the A6 cell. This intriguing result shows that paxillin, which has been characterized as a cytoskeletal protein, is capable of translocating to the nucleus. PMID- 11418192 TI - Identification and cloning of Lmairk, a member of the Aurora/Ipl1p protein kinase family, from the human protozoan parasite Leishmania. AB - Lmairk, a gene encoding a member of the Aurora/Ipl1p family of protein kinases (AIRK), was cloned from the protozoan parasite Leishmania major. Aurora kinases are key enzymes involved in the regulation of normal chromosome segregation during mitosis and cytokenesis of eukaryotic cells. This single-copy gene located on L. major chromosome 28 encodes a 301 amino acid polypeptide. All 11 conserved eukaryotic protein kinase catalytic subdomains are present and the proposed AIRK signature sequence was identified in the activation loop between subdomains VII and VIII. Lmairk is expressed, as an approximately 2.4 kb message, in at least three different species of Leishmania. This report represents the first identification of an AIRK from the trypanosomatid family of early divergent eukaryotes. PMID- 11418193 TI - Molecular identification and sequence analysis of Hillarin, a novel protein localized at the axon hillock. AB - The monoclonal antibody Lan3-15 identifies a novel protein, Hillarin, that is localized to the axon hillock of leech neurons. Using this antibody we have identified a full length cDNA coding for leech Hillarin and determined its sequence. The gene encodes a 1274 residue protein with a predicted molecular mass of 144013 Da. Data base searches revealed that leech Hillarin has potential orthologues in fly and nematode and that these proteins share two novel protein domains. The W180 domain is characterized by five conserved tryptophans whereas the H domains share 21 invariant residues. In contrast to the arrangement in fly and nematode the cassette containing the W180 and H domains is repeated twice in leech Hillarin. This suggests that the leech Hillarin sequence originated from a duplication event of an ancestral protein with single cassette structure. PMID- 11418194 TI - Is plant gene expression regulated globally? PMID- 11418195 TI - Polyphyletic origins of bacterial Nramp transporters. AB - The redox-active metals iron and manganese are required for energy metabolism, protection against oxidative stress and defense against infections. In eukaryotes, both divalent metals are transported by Nramp transporters. The sequence of these transporters was remarkably conserved during evolution. Several bacterial Nramp homologs (MntH) are also proton-dependent manganese transporters. Here, we present phylogenetic evidence for the polyphyletic origins of three groups of MntH proteins and for possible Nramp horizontal gene transfer with eukaryotes. We propose that the evolution of the MntH/Nramp family is related to adaptation to oxidative environments, including those arising during infection of animals and plants. PMID- 11418196 TI - Evolutionary expansion of CRIB-containing Cdc42 effector proteins. AB - Cdc42, a small GTPase, regulates actin polymerization and other signaling pathways through interaction with many different downstream effector proteins. Most of these effector proteins contain a Cdc42-binding domain, called a CRIB domain. Here, we describe the evolutionary analysis of these CRIB-containing proteins in yeast, worms, flies and humans. The number of CRIB-containing effector proteins increases from yeast to humans, involving both an increase within families and the emergence of new families. These evolutionary changes correlate with the development of the more complex signaling pathways present in higher organisms. PMID- 11418197 TI - Selective constraint in intergenic regions of human and mouse genomes. AB - We aligned and analyzed 100 pairs of complete, orthologous intergenic regions from the human and mouse genomes (average length approximately 12 000 nucleotides). The alignments alternate between highly similar segments and dissimilar segments, indicating a wide variation of selective constraint. The average number of selectively constrained nucleotides within a mammalian intergenic region is at least 2000. This is threefold higher than within a nematode intergenic region and at least twofold higher than the number of selectively constrained nucleotides coding for an average protein. Because mammals possess only two- to threefold more proteins than Caenorhabditis elegans, the higher complexity of mammals might be primarily because of the functioning of intergenic DNA. PMID- 11418217 TI - How do mitochondrial genes get into the nucleus? AB - It is well known that genes from chloroplasts and mitochondria were transferred to the nucleus many times during plant evolution. But in what form do the transferred genes physically make that intracellular journey--as RNA, as cDNA, as pieces of organelle DNA, or as whole organelle chromosomes? Current views focus upon cDNA as the vehicle, based upon some examples from plants. But other mechanisms, involving direct transfer of DNA from organelle chromosomes, could also account for the available data. Direct DNA transfer, rather than cDNA mediated transfer, does occur today, and it probably prevailed during the early phases of organelle evolution. PMID- 11418218 TI - Genetical genomics: the added value from segregation. AB - The recent successes of genome-wide expression profiling in biology tend to overlook the power of genetics. We here propose a merger of genomics and genetics into 'genetical genomics'. This involves expression profiling and marker-based fingerprinting of each individual of a segregating population, and exploits all the statistical tools used in the analysis of quantitative trait loci. Genetical genomics will combine the power of two different worlds in a way that is likely to become instrumental in the further unravelling of metabolic, regulatory and developmental pathways. PMID- 11418219 TI - SIR2 and aging--the exception that proves the rule. AB - One of the holy grails of medicine is the possibility of an increase in lifespan without a decrease in vitality. However, the causes and processes of human aging are still unclear. One evolutionary theory is that in the post-reproductive stage of life, selective forces decline allowing many vital systems to deteriorate. This suggests that intervention will be difficult, if not impossible. However, molecular geneticists propose an aging process that is programmed (like other developmental processes) and regulated by single genes, meaning that intervention could be possible. Here, I discuss a way of reconciling these two views that could have major implications for healthcare. PMID- 11418220 TI - Mating in mushrooms: increasing the chances but prolonging the affair. AB - Finding a compatible mating partner is an essential step in the life cycle of most sexually reproducing organisms. Fungi have two or more mating types, and only cells of different mating type combine to produce diploid cells. In mushrooms, this is taken to extremes, with the occurrence of many thousands of mating types. But, having gone to such extraordinary lengths to ensure that almost any two mushroom mycelia in the wild can mate, cell fusion is not followed by nuclear fusion and true diploidy. Instead, the fused cells form a characteristic mycelium, known as the dikaryon, in which haploid nuclei are paired but actively prevented from fusing. The mating-type genes, which encode pheromones, pheromone receptors and homeodomain transcription factors, have crucial roles in regulating the complex developmental programme by which the dikaryon is formed. PMID- 11418221 TI - Natural selection and the evolution of mtDNA-encoded peptides: evidence for intergenomic co-adaptation. AB - Mitochondrial DNA (mtDNA) variation is an important tool for the investigation of the population genetics of animal species. Recently, recognition of the role of mtDNA mutations in human disease has spurred increasing interest in the function and evolution of mtDNA and the 13 polypeptides it encodes. These proteins interact with a large number of peptides encoded in the nucleus to form the mitochondrial electron transport system (ETS). As the ETS is the primary energy generation system in aerobic metazoans, natural selection would be expected to favor mutations that enhance ETS function. Such mutations could occur in either the mitochondrial or nuclear genes encoding ETS proteins and would lead to positive intergenomic interactions, or co-adaptation. Direct evidence for intergenomic co-adaptation comes from functional studies of systems where nuclear mitochondrial DNA combinations vary naturally or can be manipulated experimentally. PMID- 11418222 TI - SNP association studies in Alzheimer's disease highlight problems for complex disease analysis. AB - Genetic linkage and association analyses are two distinct approaches to understanding the genetic etiology of complex disease. Association analysis has become particularly popular in recent times, but the true utility of the strategy remains uncertain. To try to gain better insight into the relevant issues, we have used genetic association analysis to explore the etiology of Alzheimer's disease. Our empirical findings supplement the theoretical debate, illustrating the general doubtfulness of previous positive findings and the limited ability of typical association studies based on candidate genes to discern true medium-sized signals from false positives. Improvements in genotyping technologies and increasing the number of SNPs tested, without sophisticated allowance for all other issues, could simply lead to an unmanageable overload of false-positive signals, themselves obscuring true disease associations. PMID- 11418223 TI - Evolution, language and analogy in functional genomics. AB - Almost a century ago, Wittgenstein pointed out that theory in science is intricately connected to language. This connection is not a frequent topic in the genomics literature. But a case can be made that functional genomics is today hindered by the paradoxes that Wittgenstein identified. If this is true, until these paradoxes are recognized and addressed, functional genomics will continue to be limited in its ability to extrapolate information from genomic sequences. PMID- 11418224 TI - Alcoholic myopathy: biochemical mechanisms. AB - Between one- and two-thirds of all alcohol abusers have impairment of muscle function that may be accompanied by biochemical lesions and/or the presence of a defined myopathy characterised by selective atrophy of Type II fibres. Perturbations in protein metabolism are central to the effects on muscle and account for the reductions in muscle mass and fibre diameter. Ethanol abuse is also associated with abnormalities in carbohydrate (as well as lipid) metabolism in skeletal muscle. Ethanol-mediated insulin resistance is allied with the inhibitory effects of ethanol on insulin-stimulated carbohydrate metabolism. It acutely impairs insulin-stimulated glucose and lipid metabolism, although it is not known whether it has an analogous effect on insulin-stimulated protein synthesis. In alcoholic cirrhosis, insulin resistance occurs with respect to carbohydrate metabolism, although the actions of insulin to suppress protein degradation and stimulate amino acid uptake are unimpaired. In acute alcohol dosing studies defective rates of protein synthesis occur, particularly in Type II fibre-predominant muscles. The relative amounts of mRNA-encoding contractile proteins do not appear to be adversely affected by chronic alcohol feeding, although subtle changes in muscle protein isoforms may occur. There are also rapid and sustained reductions in total (largely ribosomal) RNA in chronic studies. Loss of RNA appears to be related to increases in the activities of specific muscle RNases in these long-term studies. However, in acute dosing studies (less than 1 day), the reductions in muscle protein synthesis are not due to overt loss of total RNA. These data implicate a role for translational modifications in the initial stages of the myopathy, although changes in transcription and/or protein degradation may also be superimposed. These events have important implications for whole-body metabolism. PMID- 11418225 TI - Fluoxetine treatment of cocaine-dependent patients with major depressive disorder. AB - Sixty-eight male and female individuals with both DSM-IV diagnoses of cocaine dependence and major depressive disorder were randomly assigned to one of two medication conditions (placebo vs. 40 mg per day) as part of a double-blind, placebo-controlled clinical efficacy trial of fluoxetine for the treatment of this dual diagnosis. During the 12-week outpatient treatment phase all participants also received individual cognitive-behavioral psychotherapy targeting both cocaine use and depression. Depressive symptoms remitted as a function of time in treatment, with no significant medication effects found. Fewer cocaine positive urines were found during the first 6 weeks of treatment in the placebo group compared with the 40-mg group. Cocaine use and depressive symptoms during treatment were significantly correlated. The findings fail to support the role of fluoxetine for treatment of cocaine use and depression in dually-diagnosed patients. PMID- 11418226 TI - Sex differences in opioid antinociception: kappa and 'mixed action' agonists. AB - A number of investigators have shown that male animals are more sensitive than females to the antinociceptive effects of mu-opioid agonists. The present study was conducted to examine sex differences in opioid antinociception in the rat using agonists known to differ in selectivity for and efficacy at kappa- versus mu-receptors. Dose- and time-effect curves were obtained for s.c. U69593, U50488, ethylketazocine, (-)-bremazocine, (-)-pentazocine, butorphanol and nalbuphine on the 50 or 54 degrees C hotplate and warm water tail withdrawal assays; spontaneous locomotor activity was measured 32-52 min post-injection in the same rats. On the hotplate assay, only butorphanol (54 degrees C) and nalbuphine (50 degrees C) were significantly more potent in males than females. On the tail withdrawal assay, all agonists were significantly more potent or efficacious in males than females at one or both temperatures. In contrast, no agonist was consistently more potent in one sex or the other in decreasing locomotor activity. Estrous stage in female rats only slightly influenced opioid effects, accounting for an average of 2.6% of the variance in females' antinociceptive and locomotor responses to drug (50 degrees C experiment). These results suggest that (1) sex differences in antinociceptive effects of opioids are not mu-receptor dependent, as they may occur with opioids known to have significant kappa receptor-mediated activity; (2) the mechanisms underlying sex differences in kappa-opioid antinociception may be primarily spinal rather than supraspinal; (3) sex differences in antinociceptive effects of opioid agonists are not secondary to sex differences in their sedative effects. PMID- 11418227 TI - Prevalence and correlates of substance use disorders among older adolescents in Puerto Rico and the United States: a cross-cultural comparison. AB - Although there are substantial data on adolescent substance use, little systematic research has studied the prevalence of adolescent substance disorders in general populations, let alone cultural differences in disorder prevalence. In this paper we report the prevalence and correlates of alcohol and drug use and disorder among older adolescents on Puerto Rico (PR) and in the United States (US). Data come from an island-wide survey of the PR general residential population (15-18 year old subsample, unweighted N=922) fielded in 1997, and from the National Comorbidity Survey of the US household population (15-18 year old subsample, unweighted N=641), fielded in 1991. Both surveys used a similar standardized interview based on the Composite International Diagnostic Interview (CIDI) to generate DSM-IV diagnoses. Bivariate analyses and logistic regression were used. The major findings are: (1) US youth report higher rates of lifetime alcohol and drug use and substance-related disorder than PR youth; (2) the transition to abuse or dependence occurs for one-third of US drinkers, one-fifth of drinkers in PR, and about one-half of the drug users in both US and PR; (3) there are marked variations across sites in the types of symptoms substance users are likely to experience; (4) with the exception of family income, most correlates operate similarly in both sub-samples; and (5) the majority of both US and PR youth with a past year substance use disorder did not report any service utilization in the past year. PMID- 11418228 TI - Aggressive behavior and opportunities to purchase drugs. AB - Robins, Kellam, and others found robust evidence linking youthful aggression and deviance to later illicit drug use. Some investigators favor the interpretation that drug use is just one manifestation or complication of a more general problem behavior syndrome or conduct disorder. In this work, we test the complementary hypothesis that aggressive youth are more likely to be approached with offers to buy drugs, and found the most aggressive youths were about five times more likely to be offered drugs for purchase. However, this association was much attenuated when levels of delinquency were taken into account. In this respect, delinquent rather than aggressive behavior might be more salient. This study's evidence does not contradict previous problem behavior theories, but rather prompts new ideas about how aggression, delinquency, and drug use might be linked. One testable hypothesis is that youths with both aggression and delinquency are more likely to enter microenvironments where drug dealing is more prevalent. Or, their observable behaviors or physical appearance might function as signs of apparent willingness to try drugs. These results add to our understanding of links between aggression, delinquency, and drug use, and introduce a new line of epidemiological inquiry focused upon drug purchase opportunities. PMID- 11418229 TI - Buprenorphine for opiate addiction: potential economic impact. AB - This study evaluated the potential economic impact of the buprenorphine/naloxone combination in the context of practice in the United States of America. In comparison to treatment provided through methadone clinics, buprenorphine/naloxone therapy in office practice may be associated with increased medication, physician, and nursing costs, but reduced costs for dispensing, toxicology screens, counseling and administration. It may also result in markedly reduced costs for patients, especially travel costs, resulting in net savings for society as a whole. A review of controlled studies suggest that buprenorphine/naloxone is not likely to be any more or less effective than methadone, but since it will be less expensive in the long run, it may be more cost-effective than methadone when provided to comparable groups of patients. Because of the convenience of office-based treatment, buprenorphine/naloxone may increase access to opiate substitution for some addicts. To the extent that treatment is provided to additional high-cost patients who are involved in extensive criminal activity or who undergo multiple detoxifications each year, net cost savings could be substantial. To the extent that treatment is extended to better adjusted addicts who are employed, married and experience fewer adverse effects from their addiction, costs could increase. The total cost impact will depend on which addict sub-populations make greatest use of the treatment opportunity presented by buprenorphine/naloxone. PMID- 11418230 TI - Relationship between central serotonergic neurotransmission and reduction in alcohol intake by citalopram. AB - The relationship between the effect of citalopram on alcohol intake and central serotonergic neurotransmission, as assessed by prolactin (PRL) response to fenfluramine, was investigated in 17 male heavy drinkers. A positive correlation was obtained, suggesting that the status of central serotonergic neurotransmission in individuals is associated with the treatment response to citalopram. When the group of subjects were divided into those with high and low PRL response (above and below median, respectively) to fenfluramine, those with high PRL response had a significant reduction in alcohol intake during citalopram treatment, whereas those with low PRL response had no such effect. Thus, in subjects with evidence of unimpaired or only slightly impaired central serotonergic neurotransmission (high PRL response) citalopram may have beneficial effect on alcohol consumption, whereas in those with more evidently impaired serotonergic neurotransmission (low PRL response) citalopram treatment may have no effect on or may even increase the alcohol consumption. PMID- 11418231 TI - Craving predicts use during treatment for methamphetamine dependence: a prospective, repeated-measures, within-subject analysis. AB - Clinical lore dictates that craving drives the compulsive use of drugs and alcohol - the core feature of substance dependence. Yet limited research has yielded mixed results, suggesting that craving is neither necessary nor sufficient for continued use or relapse to addictive substances. To investigate the role of craving in compulsive methamphetamine use, 31 men and women in treatment for methamphetamine dependence were asked to indicate, once each week for 12 weeks, the severity of craving that they had experienced during the previous 24 h, using a 100-mm visual analog scale. In a prospective, repeated measures, within-subject analysis, craving intensity significantly predicted methamphetamine use in the week immediately following each craving report. Craving remained a highly significant predictor in multivariate models controlling for pharmacological intervention, and for methamphetamine use during the prior week. Craving scores that preceded use were 2.7 times higher than scores that preceded abstinence. Risk of subsequent use was 2.5 times greater for scores in the upper half of the scale relative to scores in the lower half. The results obtained demonstrate that, while craving alone may be neither necessary nor sufficient to explain substance addiction, when measured prospectively in a carefully-designed study craving emerges as a salient predictive factor in continued methamphetamine use for patients in treatment for methamphetamine dependence. PMID- 11418232 TI - A longitudinal study of the order of onset of alcohol dependence and major depression. AB - Alcohol dependence and major depression commonly occur together; however, few studies have assessed prospectively the magnitude of the risk that one disorder imparts on the subsequent occurrence of the other. We used data from the first two waves of the Epidemiologic Catchment Area community survey (n=14480) to estimate the odds of either major depression or alcohol dependence being followed by the other disorder after 1 year of follow-up. The odds of developing major depression associated with low, medium, and high levels of alcoholic symptoms at baseline were 1.66, 3.98, and 4.32 for females (P<0.001), and 1.19, 2.49, and 2.12 for males (P=0.026). Conversely, odds ratios indicating the 1-year follow-up risk of incident alcohol dependence within low, medium, and high categories of baseline depressive symptomatology were 2.75, 3.52, and 7.88 for females (P<0.001) and 1.50, 1.41, and 1.05 for males (P=0.091). Individuals with alcohol dependence appeared more likely to meet lifetime diagnostic criteria for both disorders after 1 year than individuals with depression. These results suggest that both alcohol dependence and major depression pose a significant risk for the development of the other disorder at 1 year. PMID- 11418233 TI - Regulation of herpes simplex virus gene expression. AB - Expression of the more than 80 individual genes of herpes simplex virus 1 (HSV-1) takes place in a tightly regulated sequential manner that was first described over 20 years ago. Investigations since that time have focused on understanding the mechanisms that regulate this orderly and efficient expression of viral genes. This review examines recent findings that have shed light on how this process is regulated during productive infection of the cell. Although the story is still not complete, several aspects of HSV gene expression are now clearer as a result of these findings. In particular, several new functions have recently been ascribed to some of the known viral regulatory proteins. The results indicate that the viral gene expression is regulated through transcriptional as well as post-transcriptional mechanisms. In addition, it has become increasingly clear that the virus has evolved specific functions to interact with the host cell in order to divert and redirect critical host functions for its own needs. Understanding the interactions of HSV and the host cell during infection will be essential for a complete understanding of how viral gene expression is regulated. Future challenges in the field will be to develop a complete understanding of the mechanisms that temporally regulate virus gene expression, and to identify and characterize the relevant interactions between the virus and the distinctive cell types normally infected by the virus. PMID- 11418234 TI - Gene expression patterns in the maize caryopsis: clues to decisions in embryo and endosperm development. AB - We will describe gene expression patterns in the maize caryopsis, which provide clues to developmental decisions and questions in the embryo and endosperm. The emphasis will be on the development of the root/shoot axis, which is the main achievement of plant embryogenesis. Data obtained in the vegetative seedling are included as far as they may be relevant to the elaboration of the shoot/root axis. Development of the embryo will be briefly compared to endosperm as both seed compartment exhibit pronounced differences. PMID- 11418235 TI - Osteocalcin is incompletely spliced in non-osseous tissues. AB - Osteocalcin (OC) is known to be a bone tissue-specific protein, expression of which is believed to be controlled by the OC promoter. In this communication, we provided evidence to demonstrate that tissue-specific expression of OC was also regulated at the RNA splicing level. We identified incompletely spliced variants of human OC mRNA, which retain one or more introns during RNA splicing, existing dominantly in non-osseous organs. Northern blot analysis identified two OC RNA transcripts expressed in normal human tissues, but the expression level of the transcripts varied between the tissues. Most non-osseous tissues expressed transcripts with higher molecular weight, prominent in ovary, kidney, pancreas, spleen, thymus, prostate, and testis, than the expected size of OC mRNA as seen in bone marrow. RT-PCR analysis identified up to six OC transcripts in most tissues tested except bone marrow. Sequence analysis showed that four of five RNA variants contained intron 1 in common and the dominant one contained all three introns. MG63, an osteoblastic osteosarcoma cell, expressed only the completely spliced form of OC, whereas incompletely spliced RNA was dominant in most prostate tumor cells. Combined study of in situ hybridization and immunohistochemistry revealed that OC RNA was highly expressed in prostate tumor epithelial cells while only very low levels of protein were detected, which confirms that there are OC RNA variants in non-osseous tissues. In conclusion, we demonstrated that OC mRNA is also expressed in several non-osseous tissues. However, only bone preferentially underwent the complete splicing event of all three introns. The function of other splicing variants of OC mRNA needs to be further investigated. PMID- 11418236 TI - Sox18 expression in blood vessels and feather buds during chicken embryogenesis. AB - Sox18 encodes a transcription factor known to be important for the development of blood vessels and hair follicles in mice. In order to study the functional conservation of this gene through evolution, we have isolated and characterized Sox18 in chickens. cSox18 shows a high degree of sequence homology to both the mouse and human orthologues, particularly in the high mobility group DNA-binding domain and to a lesser extent in the transcriptional activation domain. A region of unusually high sequence conservation at the C-terminus may represent a further, previously unrecognized functional domain. Both the chicken and human proteins appear to be truncated at the N-terminus relative to mouse SOX18. In situ hybridization analyses showed expression in the developing vasculature and feather follicles, consistent with reported expression in the mouse embryo. In addition, cSox18 mRNA was observed in the retina and claw beds. PMID- 11418237 TI - Isolation of hNap1BP which interacts with human Nap1 (NCKAP1) whose expression is down-regulated in Alzheimer's disease. AB - We previously reported the isolation of a novel apoptosis-related gene, human Nap1 (HGMW-approved symbol NCKAP1), the expression of which was strongly down regulated in sporadic Alzheimer's disease (AD). Human Nap1 proved to be an orthologue of rat Nap1 which binds to the adaptor molecule Nck in signal transduction. In order to further elucidate the function of human Nap1, we performed yeast two-hybrid screening. As a result of screening, we discovered a protein designated hNap1BP (human Nap1 binding protein) which is a member of the tyrosine kinase-binding protein family. In addition, hNap1BP bound to the SH3 domain of c-Abl and Nck. hNap1BP is expressed ubiquitously in various tissues like human Nap1, and intriguingly these genes are co-expressed in hippocampus and cerebral cortex in mouse brain where AD pathological features are strongly evident. Further functional analysis of hNap1BP may clarify its contribution to AD pathology. PMID- 11418238 TI - Identification of a new fibroblast growth factor receptor, FGFR5. AB - A novel fibroblast growth factor receptor (FGFR), designated FGFR5, was identified from an EST database of a murine lymph node stromal cell cDNA library. The EST has approximately 32% identity to the extracellular domain of FGFR1-4. Library screening with this EST identified two full-length alternative transcripts which we designated as FGFR5 beta and FGFR5 gamma. The main difference between these transcripts is that FGFR5 beta contains three extracellular Ig domains whereas FGFR5 gamma contains only two. A unique feature of FGFR5 is that it does not contain an intracellular tyrosine kinase domain. Predictive structural modelling of the extracellular domain of FGFR5 gamma suggested that it was a member of the I-set subgroup of the Ig-superfamily, consistent with the known FGFRs. Northern analysis of mouse and human FGFR5 showed detectable mRNA in a broad range of tissues, including kidney, brain and lung. Genomic sequencing identified four introns but identified no alternative transcripts containing a tyrosine kinase domain. Extracellular regions of FGFR5 beta and 5 gamma were cloned in-frame with the Fc fragment of human IgG(1) to generate recombinant non-membrane bound protein. Recombinant FGFR5 beta Fc and R5 gamma Fc demonstrated specific binding to the ligand FGF-2, but not FGF-7 or EGF. However, biological data suggest that FGF-2 binding to these proteins is with lower affinity than its cognate receptor FGFR2C. The above data indicate that this receptor should be considered as the fifth member of the FGFR family. PMID- 11418239 TI - Monitoring of the subtraction process in solid-phase representational difference analysis: characterization of a candidate drug. AB - In this study, we have applied and evaluated a modified cDNA representational difference analysis (RDA) protocol based on magnetic bead technology to study the molecular effects of a candidate drug (N,N'-diacetyl-L-cystine, DiNAC) in a model for atherosclerosis. Alterations in a gene expression profile induced by DiNAC were investigated in a human monocytic cell line (THP-1) differentiated into macrophage-like cells by lipopolysaccharide and further exposed to DiNAC. Three rounds of subtraction have been performed and the difference products from the second and third rounds have been characterized in detail by analysis of over 1000 gene sequences. Two protocols for analysis of the subtraction products have been evaluated, a shotgun approach and size selection of both distinct fragments and band-patterned smear. We demonstrate that in order to obtain a representative view of the most abundant gene fragments, the shotgun procedure is preferred. The obtained sequences were analyzed against the UniGene and Expressed Gene Anatomy Database (EGAD) databases and the results were visualized and analyzed with the ExProView software enabling rapid pair-wise comparison and identification of individual genes or functional groups of genes with altered expression levels. The identified differentially expressed gene sequences were comprised of both genes with known involvement in atherosclerosis or cholesterol biosynthesis and genes previously not implicated in these processes. The applicability of a solid phase shotgun RDA protocol, combined with virtual chip monitoring, results in new starting points for characterization of novel candidate drugs. PMID- 11418240 TI - Mitochondrial sequence migrated downstream to a nuclear V-ATPase B gene is transcribed but non-functional. AB - A promiscuous nuclear sequence containing a mitochondrial DNA fragment was isolated from rice. Nucleotide sequence analysis reveals that the cDNA clone #21 carries a mitochondrial sequence homologous to the 3' portion of the rps19 gene followed by the 5' portion of the rps3 gene. The mitochondrial sequence is present in an antisense orientation. Sequence comparison of the #21 cDNA with the original mitochondrial sequence shows 99% similarity, suggesting a recent transfer event. Moreover, evidence for a lack of an RNA editing event and retaining of the group II intron sequence strongly suggests that the sequence was transferred from mitochondrion to the nucleus via DNA rather than RNA as an intermediate. The upstream region to the mitochondria-derived sequence shows homology to part of the vacuolar H(+)-ATPase B subunit (V-ATPase B) gene. Isolation of a functional V-ATPase B cDNA and its comparison with the #21 cDNA reveal a number of nucleotide substitutions resulting in many translational stop codons in the #21 cDNA. This indicates that the #21 cDNA sequence is not functional. Analysis of genomic sequences shows the presence of five intron sequences in the #21 cDNA, whereas the functional V-ATPase B gene has 14 introns. Of these, three exons and their internal two introns are homologous to each other, suggesting a duplication event of V-ATPase B genomic DNA. The results of this investigation strongly suggest that the mitochondrial sequence was integrated in an antisense orientation into the pre-existing V-ATPase B pseudogene that can be transcribed and spliced. This represents a case of unsuccessful gene transfer from mitochondrion to the nucleus. PMID- 11418241 TI - A family of Tc1-like transposons from the genomes of fishes and frogs: evidence for horizontal transmission. AB - Tc1-like transposons are very widely distributed within the genomes of animal species. They consist of an inverted repeat sequence flanking a transposase gene with homology to the mobile DNA element, Tc1 of the nematode Caenorhabditis elegans. These elements seem particularly to infest the genomes of fish and amphibian species where they can account for 1% of the total genome. However, all vertebrate Tc1-like elements isolated so far are non-functional in that they contain multiple frameshifts within their transposase coding regions. Here I describe a Tc1-like transposon (PPTN) from the genome of a marine flatfish species (Pleuronectes platessa) which bears conserved inverted repeats flanking an apparently intact transposase gene. Closely related, although degenerate, Tc1 like transposons were also isolated from the genomes of Atlantic salmon (SSTN, Salmo salar) and frog (RTTN, Rana temporaria). Consensual nucleic acid sequences were derived by comparing several individual isolates from each species and conceptual amino acid sequences were thence derived for their transposases. Phylogenetic analysis of these sequences with previously isolated Tc1-like transposases shows that the elements from plaice, salmon and frog comprise a new subfamily of Tc1-like transposons. Each member is distinct in that it is not found in the genomes of the other species tested. Plaice genomes contain about 300 copies of PPTN, salmon 1200 copies of SSTN and frog genomes about 500 copies of RTTN. The presence of these closely related elements in the genomes of fish and frog species, representing evolutionary lines, which diverged more than 400 million years ago, is not consistent with a vertical transmission model for their distributions. PMID- 11418242 TI - Two Arabidopsis homologs of the animal trithorax genes: a new structural domain is a signature feature of the trithorax gene family. AB - Two Arabidopsis genes have been characterized as first examples of plant genes homologous to the animal trithorax genes. The Arabidopsis genes are highly similar but display different tissue and development expression patterns. One of them was ubiquitously expressed, with highest levels registered in young seedlings. The other gene was less active in all tested tissues, was not expressed in mature leaves but was highly expressed in roots. A new structural motif common to all TRX-related proteins has been identified. This new architectural element was found only in genes of multicellular species and is present in all genes belonging to the trithorax family. Along with the SET domain and the PHD fingers, this new element is a signature feature for the trithorax gene family. PMID- 11418243 TI - Characterization of the cDNA encoding the 90 kDa heat-shock protein in the Lepidoptera Bombyx mori and Spodoptera frugiperda. AB - This report presents the first hsp90 complete cDNA sequences from two Lepidoptera. The Bombyx mori full sequence was reconstituted from 15 partial cDNA clones belonging to expressed sequence tag libraries obtained from different tissues or cultured cells, thus showing the ubiquitous expression of the hsp90 gene. The Spodoptera frugiperda cDNA was isolated as a full-length clone from a cDNA library established from the Sf9 cell line. Both cDNAs are highly homologous and display the classical amino acid (aa) stretches representing the HSP90 signature. They potentially encode a 716 aa (B. mori) and a 717 aa (S. frugiperda) protein, with a calculated molecular mass of 83 kDa similar to the Drosophila homologous protein. We show that, unlike the vertebrates, hsp90 is a unique gene in both S. frupiperda and B. mori genomes. Sequencing of the corresponding genomic region shows that, contrary to the dipteran homologous gene, the lepidopteran hsp90 gene does not display any intron. Phylogenetic analysis based on the two lepidopteran and 23 other HSP90 aa sequences displays a high consistency with known phylogeny at both high and low taxonomic levels. Transcriptional analysis performed in S. frugiperda shows that the induction of the hsp90 gene only occurs 14 degrees C above physiological growth conditions (42 degrees C). PMID- 11418244 TI - Diptericin-like protein: an immune response gene regulated by the anti-bacterial gene induction pathway in Drosophila. AB - Insects produce various anti-microbial peptides in response to injury and infection. In Drosophila, diptericin has previously been studied as an anti bacterial immune response gene. Here, we report the cloning of the diptericin like protein (dptlp) gene as a paralog of Drosophila diptericin. By comparison of their sequences, we found that the dptlp gene has all of the functional domains conserved in the diptericin gene and other anti-bacterial proteins. The dptlp gene was rapidly induced by bacterial infections and showed different time dependent gene expression patterns from those of diptericin. Like diptericin, dptlp was specifically produced from the fat body, and its expression was strictly dependent on bacterial infections. In addition, the dptlp gene expression was almost completely abolished in the imd mutant, which implicates that its expression is regulated by the anti-bacterial arm of the Drosophila innate immune regulatory pathways. In support of this, we found GATA, interferon consensus responding element, and kappa B binding sites, which is known to be important for the proper expression of anti-bacterial genes, in the proximal promoter region of the dptlp gene. Taken together, our findings support that dptlp is a novel anti-bacterial peptide whose expression is regulated by the anti bacterial immune response mechanism. PMID- 11418245 TI - The human gephyrin (GPHN) gene: structure, chromosome localization and expression in non-neuronal cells. AB - Gephyrin was first described as a peripheral membrane protein of 93 kDa anchoring the glycine receptor (GlyR) to subsynaptic microtubules and cytoskeleton. Analysis of knock-out mice demonstrated that gephyrin has additional functions in GABA(A) receptor localization at the synapse and in the biosynthetic pathway of the molybdenum cofactor (Moco). Here we describe a human non-neuronal gephyrin cDNA and the exon/intron organization of the human gephyrin gene. We found the coding region to consist of 27 exons and to span approximately 800 kb on the long arm of chromosome 14. This structure is almost identical to that of the mouse gephyrin gene except that sequences corresponding to three exons described in rat and mouse could not be identified in human. Mutations of the GlyR subunits and of gephyrin lead to severe neuromotor phenotypes in human and mouse. Hyperekplexia involves most frequently a mutation in the GlyR alpha1 subunit in humans. However, inactivation of the Moco biosynthesis pathway results in very similar symptomatology. The recent characterization of a deletion of two exons of the gephyrin gene in a patient with symptoms typical of Moco deficiency confirmed that the involvement of gephyrin in these pathologies cannot be excluded. The precise localization of the gephyrin gene allowed us to exclude it from being a candidate for the autosomal dominant spastic paraplegia, the locus of which maps to 14q between markers D14S259 and D14S1018. A description of its structure and exon boundaries should lay the groundwork for further analysis of its expression in humans. PMID- 11418246 TI - The human GPI1 gene is required for efficient glycosylphosphatidylinositol biosynthesis. AB - The first step in glycosylphosphatidylinositol (GPI) membrane anchor biosynthesis that is defective in paroxysmal nocturnal haemoglobinuria is mediated by an N acetylglucosaminyl transferase expressed in the endoplasmic reticulum. Six human genes encode subunits of this enzyme, namely PIG-A, PIG-C, PIG-H, PIG-P, GPI1, and DPM2. Here, the human GPI1 gene is characterised. This gene is organised into eleven exons. The locus was mapped to chromosome 16p13.3 near the haemoglobin alpha chain locus. GPI1 is expressed ubiquitously in human cells and tissues. Expression levels are markedly elevated in haematopoietic tissues (bone marrow, foetal liver). To determine whether human GPI1 is essential for human GPI biosynthesis, antisense RNA was expressed in HEK293 cells. Transfectants exhibited a marked but incomplete decrease in the expression of a GPI-linked reporter protein, confirming that GPI1 is required for efficient GPI biosynthesis. In contrast, expression of GPI-linked proteins is normal in lymphatic cell lines from individuals with the alpha thalassaemia/mental retardation syndrome, which is characterised by large deletions from chromosome 16p removing one of the two GPI1 alleles along with the haemoglobin alpha locus. In conclusion, GPI1 plays an important role in the biosynthesis of GPI intermediates. Due to its autosomal localisation, the heterozygous deletion of GPI1 does not lead to an overt defect in the expression of GPI-linked proteins. PMID- 11418247 TI - Heterogeneity of mRNA coding for Caenorhabditis elegans coronin-like protein. AB - We analyzed transcripts coding for the nematode Caenorhabditis elegans coronin, which had been identified by the genome project of C. elegans. We found that the gene coding for the C. elegans coronin has an alternatively spliced exon containing an alternative 5' splice site in the 3'-region. Moreover, two exons are internally cleaved by a mechanism different from the conventional splicing rules. In consequence, the gene produces five kinds of transcripts. PMID- 11418248 TI - Cloning of a novel 2',5'-oligoadenylate synthetase-like molecule, Oasl5 in mice. AB - The 2',5'-oligoadenylate synthetase (2-5OAS) is a enzyme that catalyzes synthesis of 2',5'-oligoadenylates (2-5A) in a dsRNA-dependent manner, and known as a major component of the IFN-induced host defense mechanisms against microbial infections. Here, we report the presence of a novel 2-5OAS-like molecule, termed Oasl5, in mice. The size of Oasl5 cDNA was about 2 kb and encoded a protein consisting of 362 aa. The amino acid sequence showed 76% similarity to the mouse 2-5OAS, however, several motifs being important for the enzyme activity were not conserved. The Oasl5 mRNA was most significantly expressed in the brain, and relatively weak expression was found in other organs such as the spleen, kidney, ovary and testis. It was also expressed in embryonic stem (ES) cells. The Oasl5 mRNA expression in ES cells was elevated 5-fold after treatment with IFN and about 2-fold in the brain when stimulated with IFN inducer, polyinosinic polycytidylic acid (poly[I:C]). In situ hybridization analysis revealed that Oasl5 is expressed in neurons in the central nervous system in adult mice. When Oasl5 was expressed in E. coli, it yielded 42 kDa protein that binds to dsRNA, but it did not show oligoadenylate synthetase activity. These findings suggest a novel function of Oasl5, which are independent of oligoadenylate synthetase activity, in the brain and developing embryos. PMID- 11418249 TI - Species identity: conserved inverted LINE repeat clusters (ILRC) in the vertebrate genome as indicators of population boundaries. AB - The avian LINE CR1 generates multiply-superimposed insertions, resulting in apparently fortuitous inverted LINE repeat clusters (ILRCs). These loci display size micro-heterogeneity within populations, with few or no presence/absence polymorphisms, and yet only very closely related species share loci. Whilst the CR1 sequences that the ILRC loci are derived from are not species-specific, the loci themselves appear to be species-characteristic if not totally species specific. The mammalian LINE L1 is shown to act similarly to CR1 and also forms ILRCs. It is proposed that whilst the formation of these loci may be from a near random process of super-insertion certain of them are in some way functional, explaining their conservation and rapid spread to population boundaries, whilst non-functional or inactive loci are quickly lost. ILRCs appear to decay from the element as formed by the accumulation of point mutations. ILRCs appear to an unusual example of non-polymorphic sequences being younger than polymorphic sequences with no obvious selective reason. PMID- 11418250 TI - Characterization of two genes, Impa1 and Impa2 encoding mouse myo-inositol monophosphatases. AB - The enzyme myo-inositol monophosphatase (Impa) catalyzes the synthesis of free myo-inositol from various myo-inositol monophosphates in the phosphatidylinositol signaling system. Impa is a lithium-blockable enzyme that has been hypothesized to be the biological target for lithium-salts used as mood-stabilizing drugs in the treatment of manic-depressive (bipolar) illness. As an initial step to explore the functional consequences of reduced or absent Impa activity in an animal model we here report the isolation of two Impa-encoding mouse genes, Impa1 and Impa2. Impa1 spans approximately 17.5 kb and contains nine exons of 46--1354 bp encoding a protein of 277 amino acids. Impa2 spans at least 19.5 kb and contains eight exons of 46--444 bp size encoding a protein of 290 amino acids. The genomic structure including the positions of the exon-intron splice sites seems to be conserved among myo-inositol monophosphatase genes in mammalian species. One or more Impa-like genes do also exist in evolutionary more distant species like invertebrates, plants and bacteria. The proteins encoded by the non vertebrate genes seem to be equally related to Impa1 and Impa2. We therefore suggest that the Impa1 and Impa2 genes duplicated from a common ancestral gene after the evolutionary divergence of vertebrates. PMID- 11418251 TI - Priorities among recommended clinical preventive services. AB - BACKGROUND: Many recommended clinical preventive services are delivered at low rates. Decision-makers who wish to improve delivery rates, but face competing demands for finite resources, need information on the relative value of these services. This article describes the results of a systematic assessment of the value of clinical preventive services recommended for average-risk patients by the U.S. Preventive Services Task Force. METHODS: The assessment of services' value for the U.S. population was based on two dimensions: burden of disease prevented by each service and cost effectiveness. Methods were developed for measuring these criteria consistently across different types of services. A companion article describes the methods in greater detail. Each service received 1 to 5 points on each of the two dimensions, for total scores ranging from 2 to 10. Priority opportunities for improving delivery rates were determined by comparing the ranking of services with what is known of current delivery rates nationally. RESULTS: The highest ranked services (scores of 7+) with the lowest delivery rates (< or =50% nationally) are providing tobacco cessation counseling to adults, screening older adults for undetected vision impairments, offering adolescents an anti-tobacco message or advice to quit, counseling adolescents on alcohol and drug abstinence, screening adults for colorectal cancer, screening young women for chlamydial infection, screening adults for problem drinking, and vaccinating older adults against pneumococcal disease. CONCLUSIONS: Decision makers can use the results to set their own priorities for increasing delivery of clinical preventive services. The methods provide a basis for future priority setting efforts. PMID- 11418252 TI - Methods for priority setting among clinical preventive services. AB - Methods used to compare the value of clinical preventive services based on two criteria-clinically preventable burden (CPB) and cost effectiveness (CE)-are described. A companion article provides rankings of clinical preventive services and discusses its uses for decision-makers; this article focuses on the methods, challenges faced, and solutions. The authors considered all types of data essential to measuring CPB and CE for services recommended by the U.S. Preventive Services Task Force and developed methods essential to ensuring valid comparisons of different services' relative value. PMID- 11418253 TI - A clinical trial of tailored office systems for preventive service delivery. The Study to Enhance Prevention by Understanding Practice (STEP-UP). AB - BACKGROUND: The potential of primary care practice settings to prevent disease and morbidity through health habit counseling, screening for asymptomatic disease, and immunizations has been incompletely met. This study was designed to test a practice-tailored approach to increasing preventive service delivery with particular emphasis on health habit counseling. DESIGN: Group randomized clinical trial and multimethod process assessment. SETTING/PARTICIPANTS: Seventy-seven community family practices in northeast Ohio. INTERVENTION: After a 1-day practice assessment, a nurse facilitator met with practice clinicians and staff and assisted them with choosing and implementing individualized tools and approaches aimed at increasing preventive service delivery. MAIN OUTCOME MEASURE: Summary scores of the health habit counseling, screening and immunization services recommended by the U.S. Preventive Services Task Force up to date for consecutive patients during randomly selected chart review days. RESULTS: A significant increase (p=0.015) in global preventive service delivery rates at the 1-year follow-up was found in the intervention group (31% to 42%) compared to the control group (35% to 37%). Rates specifically for health habit counseling (p=0.007) and screening services (p=0.048) were increased, but not for immunizations. CONCLUSIONS: An approach to increasing preventive service delivery that is individualized to meet particular practice needs can increase global preventive service delivery rates. PMID- 11418254 TI - Cost utility of public clinics to increase pneumococcal vaccines in the elderly. AB - BACKGROUND: Pneumococcal immunization has been shown to be cost effective, is recommended by the Advisory Committee on Immunization Practices, and is covered by Medicare. Despite that, over 50% of the population aged > or =65 is not vaccinated, leading to significant mortality and morbidity. The objective of this study is to evaluate the costs and the cost utility of immunization in nontraditional settings (community clinics set up to provide influenza and pneumococcal vaccinations) as a strategy to increase pneumococcal immunization rates. METHODS: A cost-utility analysis of public immunization clinics in Monroe County, New York, during the fall of 1998. The study included 1207 adults aged > or =65. Costs of operating the clinics and of vaccine administration were measured. The cost of health sequela and estimates of quality-adjusted life years (QALYs) were obtained from prior studies. Sensitivity analyses were performed to test several important assumptions. RESULTS: Unlike immunizations in physician offices, immunizations in nontraditional settings are not cost saving. Estimates of incremental cost-utility ratios ranged from $4215 per QALY to $12,617 per QALY, depending on the underlying assumptions of the model. CONCLUSIONS: Clinics in nontraditional settings offering pneumococcal immunization have cost-utility ratios near and below those of other recommended vaccines. These results suggest that such clinics should be considered a viable strategy for increasing pneumococcal immunization rates. PMID- 11418255 TI - The impact of Put Prevention into Practice on selected clinical preventive services in five Texas sites. AB - OBJECTIVE: To determine whether the implementation of the Put Prevention Into Practice (PPIP) office-based system would increase the delivery rates of specific clinical preventive services among demonstration clinics. METHODS: Chart review was conducted before (n=372) and 33 to 39 months after (n=376) the implementation of the PPIP office-based system in two community health centers and three family practice residency programs in Texas. The population included all adult patients aged > or = 19 years who had presented to the clinic during the study periods. RESULTS: Documentation of timely cholesterol screening increased from 70% to 84%; smoking assessment, from 56% to 80%; for women, up-to-date Papanicolaou smear, from 70% to 81%; annual mammograms (women aged > or =51), from 30% to 48%; and up to-date tetanus-diphtheria immunizations, from 19% to 59%. For adults aged > or =66 years, documentation of pneumococcal immunization increased from 22% to 48%, while influenza immunizations improved, although not significantly (45% to 49%). Blood pressure screening was almost universal (99%) at baseline and at 33- to 39 month follow-up. CONCLUSION: PPIP system changes were associated with an observed increase in delivery of selected clinical preventive services. PMID- 11418256 TI - Impact of an Australian mass media campaign targeting physical activity in 1998. AB - BACKGROUND: Physical activity is now a public health priority, but there is only limited evidence on the effectiveness of mass-reach campaigns. INTERVENTION: Paid and unpaid television and print-media advertising, physician mail-outs, and community-level support programs and strategies. SETTING/PARTICIPANTS: A mass media statewide campaign to promote regular moderate-intensity activity was conducted during February 1998. The target group was adults aged 25 to 60 who were motivated but insufficiently active. DESIGN: Cohort and independent-sample, cross-sectional representative population surveys, before and after the campaign. The intervention was conducted in the state of New South Wales, with the other states of Australia as the comparison region. MEASURES: Telephone survey items on physical activity, media message awareness, physical activity knowledge, self efficacy, and intentions. RESULTS: Unprompted recall of the activity messages in the campaign state increased substantially from 2.1% to 20.9% (p<0.01), with small changes elsewhere in Australia (1.2% to 2.6%). There were large changes in prompted awareness from 12.9% to 50.7% (p<0.0001), much larger than changes elsewhere (14.1% to 16%, p=0.06). Knowledge of appropriate moderate-intensity activity and physical activity self-efficacy increased significantly and only in the campaign state. Compared to all others, those in the target group who recalled the media message were 2.08 times more likely to increase their activity by at least an hour per week (95% confidence interval = 1.51-2.86). CONCLUSIONS: This integrated campaign positively influenced short-term physical activity message recall, knowledge, and behavior of the target population, compared to the population in the region who were not exposed. PMID- 11418257 TI - Parental provision of transportation for adolescent physical activity. AB - BACKGROUND: Physical activity (PA) is important to adolescents' health. Parent transportation to activity locations is a practical strategy for increasing youth PA that has rarely been examined. DESIGN: Cross-sectional surveys of students and parents. SETTING/PARTICIPANTS: Ethnically diverse students (N=1678; 712 boys, 966 girls; M age=13.0 years) from 24 middle schools (grades six to eight) and their parents completed surveys (response rate=72%). MAIN OUTCOME MEASURES: Frequency of parents transporting adolescents to PA locations was studied in relation to adolescents' reported participation in PA during the previous week and their involvement in sports and activity lessons during the past year. RESULTS: Parents transported adolescents to PA locations 2.13 times per week, with boys being transported more often than girls (p=0.03). Ethnic/racial differences in frequency of transport were evidenced (p=0.002). Parent transportation for PA significantly contributed to girls' total PA (p=0.001) and their participation in sports/activity lessons (p=0.001). Transportation contributed marginally (p=0.06) to boys' total PA, but significantly to their participation in sports/activity lessons (p=0.001). CONCLUSIONS: Parent provision of transportation to activity locations is associated with out-of-school PA in a diverse adolescent population. This variable should be targeted for intervention. PMID- 11418258 TI - Barriers to prenatal care in Europe. AB - BACKGROUND: In Europe, it is sometimes assumed that few barriers to prenatal care exist because extensive programs of health insurance and initiatives to promote participation in prenatal care have been established for many decades. METHODS: A case-control study was performed in ten European countries (Austria, Denmark, Germany, Greece, Hungary, Ireland, Italy, Portugal, Spain, and Sweden). Postpartum interviews were conducted between 1995 and 1996. A total of 1283 women with inadequate prenatal care (i.e., with 0, 1, or 2 prenatal care visits or a first prenatal care visit after 15 completed weeks of pregnancy) and 1280 controls with adequate prenatal care were included in the analysis combining data from the ten countries. RESULTS: Based on combined data of the ten countries, lack of health insurance was found to be an important risk factor for inadequate prenatal care (crude odds ratio [OR] at 95% confidence interval [CI]: 30.1 [20.1 47.1]). Women with inadequate prenatal care were more likely to be aged < 20 years (16.4% vs 4.8%) and with higher parity (number of children previously borne) than controls. They were more likely to be foreign nationals, unmarried, and with an unplanned pregnancy. Women with inadequate care were also more likely to have less education and no regular income. They had more difficulties dealing with health services organization and child care. Cultural and financial barriers were present, but after adjusting for confounders by logistic regression, perceived financial difficulty was not a significant factor for inadequate prenatal care (adjusted OR [95% CI]: 0.7 [0.4-1.3]). CONCLUSIONS: Personal, socioeconomic, organizational, and cultural barriers to prenatal care exist in Europe. PMID- 11418259 TI - A village would be nice but...it takes a long-acting contraceptive to prevent repeat adolescent pregnancies. AB - OBJECTIVE: To determine which components of a comprehensive, multidisciplinary, adolescent-oriented maternity program help teenage mothers delay subsequent pregnancies. DESIGN/SETTING/PATIENTS: A cohort of 373 participants in a comprehensive, multidisciplinary, adolescent-oriented maternity program was studied. INTERVENTION: The program was designed to prevent rapid subsequent pregnancies directly by simplifying access to contraceptives and indirectly by discouraging school drop-out and encouraging the pursuit of careers incompatible with closely spaced childbearing. Parents and children were seen together; nine visits were scheduled during the first postpartum year and four visits during the second year. MAIN OUTCOME MEASURE: Repeat adolescent pregnancy. RESULTS: The repeat pregnancy rate was 14% at 1 year and 35% at 2 years. Teenage mothers who became pregnant exhibited significantly more repeat pregnancy risk factors but were as compliant with clinic visits as their nonpregnant peers. The contraceptive choices the teenagers made during the puerperium had the most profound effect on their subsequent fertility. A logistic regression analysis identified failure to use Norplant during the puerperium as the strongest predictor of repeat pregnancy during the first 2 postpartum years (relative risk [RR]=8.89; 95% confidence interval [CI]=2.80-28.50). Exhibiting nine or more repeat pregnancy risk factors (RR=2.37; 95% CI=1.38-4.06) and not using Depo Provera during the puerperium (RR=2.30; 95% CI=1.60-3.29) also predicted repeat conception, but clinic visits and return to school postpartum did not. CONCLUSIONS: Using a long-acting hormonal contraceptive during the puerperium was associated with pregnancy prevention during the first 2 postpartum years, but frequent clinic visits, contact with supportive healthcare and social service providers, and return to school were not. PMID- 11418260 TI - Priorities among recommended clinical preventive services. PMID- 11418261 TI - STEP(ing)-UP to deliver clinical preventive services. PMID- 11418263 TI - American College of Preventive Medicine Practice Policy statement. Weight management counseling of overweight adults. AB - On the basis of a review of the current literature and recommendations, the American College of Preventive Medicine presents a practice policy statement on weight management counseling of overweight adults. PMID- 11418264 TI - Cocaine decreases cell survival and inhibits neurite extension of rat locus coeruleus neurons. AB - Cocaine use during pregnancy is affiliated with neurobehavioral abnormalities in offspring that are associated with problems of attention. Given the putative role of the noradrenergic system in attentional processes, impairments in the noradrenergic system may underlie specific attentionally sensitive, neurobehavioral alterations. Recent data using a clinically relevant intravenous (iv) route of administration show that the norepinephrine cell bodies of the locus coeruleus (LC) are a primary target for in utero cocaine exposure. Cell survival and neurite outgrowth of LC neurons were studied using two paradigms: (1) in vitro, using a physiologically relevant concentration of cocaine, and (2) in vivo, using a clinically relevant intravenous rat model. Fetal cocaine exposure significantly decreased neuronal survival (in vitro: P=.0001, n=24; in vivo: P=.0337, n=30), reduced neurite initiation (in vitro: P=.001, n=24; in vivo: P=.0169, n=30), decreased the number of neurites elaborated (in vivo: P=.0031, n=30), and reduced total neurite length (in vivo: P=.0237, n=30). The results of this novel approach toward an understanding of noradrenergic neurons as they respond to cocaine during development suggest that cocaine may affect behavior by negatively regulating neuronal pathfinding and synaptic connectivity. PMID- 11418265 TI - Prenatal exposure to organic solvents and child neurobehavioral performance. AB - The present study compared the cognitive and behavioral functioning of 3- to 7 year-old children (n=33) whose mothers worked with organic solvents during pregnancy with a group of unexposed children (n=28) matched on age, gender, parental socioeconomic status (SES), and ethnicity. Participants were recruited prospectively by the Motherisk Program, an antenatal counseling service in Canada. An exposure index was estimated using questionnaire data obtained at the time of initial contact. Groups were compared on a variety of tasks, including subtests from the NEPSY, a visual CPT, as well as on parent-rated measures of children's behavior. Regression analyses indicated lower composite scores in children with increased exposure on Receptive language (P<.01), Expressive language (P<.01), and Graphomotor ability (P=.001), adjusted for demographics. No group differences were observed on measures of Attention (P=.97), Visuo-spatial ability (P=.33), and Fine-motor ability (P=.33). On the Child Behavior Checklist (CBCL), overall mean differences on broad- and narrow-band scales were not significant, but significantly more exposed children were rated as having mild or severe problem behaviors. The findings suggest that maternal occupational exposure to organic solvents during pregnancy is associated with poorer outcome in selective aspects of cognitive and neuromotor functioning in offspring. PMID- 11418266 TI - The disposition of benzoylecgonine in maternal and fetal rats. AB - We tested our hypothesis that pregnancy alters the pharmacokinetic profile of benzoylecgonine, and that this metabolite accumulates in the fetus longer than in the mother. Chronically catheterized near-term pregnant and nonpregnant female Sprague-Dawley rats received an intravenous infusion of benzoylecgonine over a period of 30 min. Adult or fetal blood and tissue samples were obtained either at the end of the infusion or 6 h postinfusion for analysis of benzoylecgonine and other cocaine metabolite concentrations via gas chromatography/mass spectrometry (GC/MS). Pregnancy altered benzoylecgonine pharmacokinetics. At the end of the infusion, benzoylecgonine concentration in the fetal plasma was markedly lower than in the maternal plasma with a fetal/maternal ratio of 0.14+/-0.01. A significantly lower concentration of benzoylecgonine was found in both maternal and fetal brain at 0 h postinfusion, with tissue/plasma concentration ratios of 0.04 and 0.24, respectively, suggesting that benzoylecgonine does not readily penetrate into the brain. At 6 h, the fetal concentration of benzoylecgonine was significantly higher than in the corresponding maternal blood and tissues. Ecgonine methyl ester, a metabolite of benzoylecgonine was found in the maternal liver, but not in the fetus. In addition, the amniotic fluid concentration of benzoylecgonine became significantly higher in the 6-h postinfusion samples as compared to the end of infusion value, suggesting that repeated intrauterine exposure to cocaine may cause an accumulation of benzoylecgonine in the fetus. PMID- 11418267 TI - Neurobehavioral teratogenic effects of thalidomide in rats. AB - Thalidomide-induced embryopathy has been known for four decades, however, the drug has been reintroduced for human use in a number of countries, including the United States. In utero thalidomide exposure in humans is associated with central nervous system (CNS) effects in addition to the well-known limb, ear and other malformations. Despite knowledge of these CNS effects, not a single experimental study could be found that examined thalidomide for possible developmental neurobehavioral effects. In the present experiment, gravid Sprague-Dawley rats were treated with either thalidomide (100 mg/kg by gavage) or vehicle (propylene glycol) on embryonic days E7-18 and allowed to deliver and raise their own offspring. The offspring were evaluated in a series of neurobehavioral tests (reflexes, locomotor activity, startle reactivity and learning in the Morris and Cincinnati water mazes). There was a small reduction in maternal weight among thalidomide-treated dams during midgestation. Thalidomide offspring showed increased preweaning mortality and male-specific, late onset reduction in growth that persisted until the end of the study. Male thalidomide offspring showed significant increases in errors and latency in the multiple-T Cincinnati water maze. Although rats are refractory to thalidomide-induced teratogenesis, the present results suggest that thalidomide selectively impairs offspring survival and growth and at least one type of learning among male offspring. PMID- 11418268 TI - A table of color distance scores for quantitative scoring of the Lanthony Desaturate color vision test. AB - The Lanthony Desaturate Panel D-15 (D-15d) color vision test is used in neurotoxicological testing to assess acquired color vision deficits. The original test design included a qualitative scoring method. Quantitative scoring requires mapping the colored objects used in the test into a color space describing perceptual distances. A table of these distances has previously been published for the saturated version of this color vision test, but not the desaturate test. This communication includes a table of color distances for the calculation of Bowman's Total Color Distance Score (TCDS) for the D-15d. This table should be useful for non-computerized scoring under field test conditions or for devising one's own computerized scoring methods using the tabulated color distances for a look-up table. Data analysis programs using SAS or Matlab are available from the author. PMID- 11418269 TI - A historical cohort study of kidney damage in long-term lithium patients: continued surveillance needed. AB - BACKGROUND: Insufficient knowledge on the longitudinal fate of renal function in lithium patients incited this retrospective study of 149 patients. METHOD: Medical record review of a lithium cohort (N = 149), 8--12 years after an initial renal function study. RESULTS: Twenty-one patients had died, one from uremia probably not caused by lithium, and 42 had discontinued lithium. Reduced urinary concentrating capacity (Umax) or glomerular filtration rate (GFR) was not more frequent among deceased or off-lithium patients than among the 86 patients who were on lithium at follow-up. In 63 of the latter patients, Umax had been re examined after the initial study, and GFR in 29 patients. Reduced Umax and GFR had become twice as common, and average Umax and GFR had decreased significantly. The reduction of GFR was associated with lithium treatment duration and age, and reduced Umax with treatment duration only. CONCLUSIONS: Reduced renal function is not a major cause of treatment discontinuation but becomes increasingly common with treatment duration.Limitations. Missing data rendered the interpretation difficult in some respects. Clinical relevance. The increased proportion of patients with reduced GFR and Umax with time implies an increased risk of potentially lethal dehydration and lithium intoxication. Continued surveillance of urinary output and GFR is therefore necessary. PMID- 11418270 TI - The Nordic Study on schizophrenic patients living in the community. Subjective needs and perceived help. AB - In a community sample of 418 persons diagnosed with schizophrenia, subjective needs and perceived help was measured by the Camberwell Assessment of Need (CAN). The mean number of reported needs was 6.2 and the mean number of unmet needs 2.6. The prevalence of needs varied substantially between the need areas from 3.6% ('telephone') to 84.0% ('psychotic symptoms'). The rate of satisfaction estimated as the percentage of persons satisfied with the help provided within an area varied between 20.0% ('telephone') and 80.6% ('food'). The need areas concerning social and interpersonal functioning demonstrated the highest proportion of unmet to total needs. In a majority of need areas the patients received more help from services than from relatives, but in the areas of social relations the informal network provided substantial help. In general the patients reported a need for help from services clearly exceeding the actual amount of help received. In a linear regression model symptom load (BPRS) and impaired functioning (GAF) were significant predictors of the need status, explaining 30% of the variance in total needs and 20% of the variance in unmet needs. It is concluded that the mental health system fails to detect and alleviate needs in several areas of major importance to schizophrenic patients. Enhanced collaboration between the care system and the informal network to systematically map the need profile of the patients seems necessary to minimise the gap between perceived needs and received help. PMID- 11418271 TI - Has the attempt to reduce psychiatric inpatient treatment been successful in Finland? AB - The deinstitutionalisation process in Finland's psychiatric healthcare did not start until the late 1980s. Our aim is to evaluate how the use of psychiatric inpatient treatment was associated with deinstitutionalisation given the changes in the modality of treatment ideology (years 1987--1991) as well as being due to economic pressures (years 1991--1995). Special emphasis is given on the inpatient treatment of schizophrenia and other psychotic disorders. Data was retrieved using the national hospital discharge register of all treatment periods in psychiatric hospitals and treatment periods due to psychiatric disorders in other hospitals. Three years (1987, 1991, and 1995) were compared. Four healthcare districts in northern Finland were studied. Resource use was measured by number of treatment periods and inpatient days in relation to population. Psychiatric inpatient treatment was reduced in 1987--1991, when resources in community care increased. During the period of economic pressures (1991--1995), when community care resources no longer increased, inpatient treatment started to rise again. Over the whole period, psychiatric treatment in primary care institutions increased. Reduction of psychiatric beds results in increasing inpatient treatment in non-specialist institutions, especially when community care fails to serve the patients. In the future it is important to evaluate whether the quality of care remains in the standard of specialised services when treatment shifts away from the specialist level. PMID- 11418272 TI - Occurrence and clinical correlates of psychiatric co-morbidity in delusional disorder. AB - The present study investigated the occurrence and the clinical correlates of psychiatric co-morbidity in a sample of 64 patients with delusional disorder (DD). Subjects were evaluated with a semi-structured interview for the collection of demographic and clinical features of the disorder; co-morbid axis 1 disorders were determined according to the clinical interview using DSM-IV by Othmer and Othmer. Delusional disorder subjects with and without co-morbid diagnoses were compared to investigate whether the presence of another psychiatric disorder influenced the clinical features of the illness.Seventy-two percent of the subjects had at least one additional lifetime psychiatric diagnosis. High lifetime co-morbidity was found with affective disorders, whose onset generally had been subsequent to the onset of DD. Patients with at least one co-morbid disorder (N = 46) had an earlier age at onset, presented for the first psychiatric consultation at an earlier age, and were younger at index evaluation for this study with respect to patients without co-morbidity (N = 18). Types of DD differed significantly according to the presence/absence of lifetime co-morbid disorders: DD patients with co-morbidity were in most cases persecutory type (54.4%) while DD patients without co-morbidity were mixed type (66.7%). Our data indicate that there is a considerable proportion of patients whose DDr is strictly connected with other co-occurring psychiatric disorders (mainly affective disorders), which exert an influence on the phenomenology of the illness. PMID- 11418273 TI - Depression in a sample of Turkish type 2 diabetes patients. AB - Forty-one depressive and 57 nondepressive patients with non-insulin-dependent diabetes mellitus (NIDDM, type 2) were compared for demographic and clinical variables. The depressive patients were more likely to have a history of depression than non-depressive patients. The female patients were more likely to have depression than males. Results suggest that depression in NIDDM was only associated with female gender and previous depressive episodes. PMID- 11418274 TI - The prognostic value of initial EDI scores in anorexia nervosa patients: a prospective follow-up study of 5--10 years. Eating Disorder Inventory. AB - AIM: To study in a prospective manner the long-term prognostic value of the initially recorded Eating Disorder Inventory (EDI) scores in anorexia nervosa (AN) patients. METHODS: The 5--10-year outcome of 26 consecutive malnourished AN patients was prospectively recorded according to the initial EDI score. We selected only patients with full 6-month assessments for more than 5 years (mean 8.5 years). Eating behavior, quality of life, autonomy and insight capacity were prospectively assessed by the Morgan-Russell scale and a semi-structured interview. At the end of follow-up, 13 patients recovered and the 13 others had a poor outcome. RESULTS: In monovariate analyses, high initial EDI total score (P <.0007) and high initial scores for perfectionism (P <.001), ineffectiveness (P <.002), interpersonal distrust (P <.004), interoceptive awareness (P <.03) and drive for thinness (P <.05) were significantly associated with a poor prognosis 5 -10 years afterward. In a multivariate analysis, only high initial scores for perfectionism (F = 8.43; P = 0.008) and interpersonal distrust (F = 7.46; P = 0.012) were significantly associated with illness severity. DISCUSSION: High EDI total score and subscales for perfectionism and interpersonal distrust could predict a long-term severe outcome in AN. PMID- 11418275 TI - Quality of life and disability in patients with obsessive-compulsive disorder. AB - The aim of this study is to describe the situation of Spanish obsessive compulsive disorder (OCD) patients and compare it to that of the general population and other patient groups. METHODS: Thirty-six OCD patients on maintenance treatment were evaluated using the Y-BOCS, SF-36, and DAS-S. Their SF 36 scores were compared to Spanish norms and to those obtained from U.S. OCD patients, schizophrenic outpatients, depressed outpatients, heroin dependents, patients on hemodialysis, and kidney transplant recipients. RESULTS: Sixty-one percent of the patients had severe or extremely severe symptoms. Their quality of life was worse when compared with the Spanish norms in all SF-36 areas, but especially with respect to mental health. In contrast to U.S. OCD patients, social functioning is more impaired in the Spanish OCD patients. OCD patients reported the same quality of life as schizophrenics in the areas of mental health, but better in the areas of physical health. Compared with heroin dependents and depressed patients, their quality of life was worse. On mental health scales, OCD patients scored worse than somatic patients. CONCLUSIONS: OCD in the Spanish population was shown to be associated with worse quality of life than for any other patient group (including physical groups), except schizophrenics. PMID- 11418276 TI - An animal model for the study of the genetic bases of behaviour in men: the multiple marker strains (MMS). AB - Animal models are often used for preclinical research on the neurobiology of psychiatric disorders. Whereas many are employed to screen new therapeutic agents, few of them are used to study the genetic bases of psychiatric diseases, probably because of the complex genetic determinism underlying quantitative behavioral traits such as mood, personality or intelligence. The present article presents a short review introducing an analysis model using mice: the marker strains model. Using this model it is possible both to display genetic determinism data and to locate some of the chromosomal fragments involved in the regulation of anxiogenic processes. At present it cannot accurately determine the position of one or more genes, but it does provide a valuable means of 'scanning' the genome for an approximation. Through genetic analysis, using the model, an attempt will be made to identify autosomal fragments which may be involved in two behavioural traits: anxiety and chemical-induced seizures. In this paper, after reviewing theoretical aspects of looking for genes involved in behaviour, we will successively introduce studies in genetic topics in psychiatric human studies as well as appropriated behavioural animal studies. Then we will present a genetic model in mice which allows us to locate chromosomal fragments associated with a behavioural trait: multiple marker strains. PMID- 11418277 TI - Tardive dyskinesia caused by the atypical antipsychotic risperidone and cured by the use of another drug of the same class, olanzapine. PMID- 11418278 TI - Effects of selective 5-HT1A receptor antagonists on regional serotonin synthesis in the rat brain: an autoradiographic study with alpha-[14C]methyl-L-tryptophan. AB - The effects of acute and chronic administration of WAY100635 and WAY100135, serotonin (5-HT)1A antagonists, on 5-HT synthesis rates, calculated from the trapping of alpha-[14C]methyl-L-tryptophan (alpha-MTrp), were evaluated in the rat brain using autoradiography. In the acute treatment studies, WAY100635 (1 mg/kg) induced a significant increase in 5-HT synthesis in the median raphe nucleus and some nerve terminal structures (range between 18 and 53%), while WAY100135 (10 mg/kg) produced a significant decrease of synthesis, in the range between 16 and 33%, in the raphe magnus nucleus and several projection areas. The action of WAY100635 given acutely was likely a result of antagonist actions at the 5-HT1A somato-dendritic autoreceptors. WAY100135 probably acted acutely as a partial agonist. In the chronic treatment studies, WAY100635 (1 mg/kg/day) and WAY100135 (10 mg/kg/day) were administered for 7 days as s.c. injections once a day. Chronic treatment with both compounds significantly reduced the rate of 5-HT synthesis in the nerve terminal structures and produced a significant increase in the raphe nuclei. These treatments did not have any effect on the plasma free or total tryptophan. PMID- 11418279 TI - Increased serum tumor necrosis factor alpha concentrations in major depression and multiple sclerosis. AB - There is now evidence that major depression is accompanied by activation of the inflammatory response system (IRS) as indicated by an increased production of pro inflammatory cytokines. There is circumstantial evidence implicating pro inflammatory cytokines, such as tumor necrosis factor-alpha (TNFalpha) in the pathogenesis of multiple sclerosis (MS). The aims of the present study were to examine (i) the serum concentrations of interleukin-6 (IL-6), IL-8, TNFalpha, IL 2 receptor (IL-2R) and CC16 (uteroglobulin), an endogenous anti-cytokine, in depressed and MS patients compared to normal controls, and (ii) the effects of treatment with antidepressants on the above IRS variables in depressed patients. Serum TNFalpha was significantly higher in depressed and MS patients than in normal controls. Serum IL-8 was significantly higher in depressed patients than in patients with MS. Serum CC16 was significantly higher in patients with MS than in normal controls and depressed patients. Nonresponders to treatment with antidepressants had significantly higher serum IL-2R and lower serum CC16 concentrations than responders to treatment. The results show that (i) depression is accompanied by activation of the IRS and that this activation is more pronounced in depression than in MS, and (ii) IRS activation in depressed patients is related to a nonresponse to treatment with antidepressants. PMID- 11418280 TI - Placebo affects the performance of rats in models of depression: is it a good control for behavioral experiments? AB - Experimental design of behavioral studies in animals generally includes placebo treated controls. However, when placebo is administered by injection in experimental models of psychiatric diseases such as depression, where stress may affect the execution of the behavioral test, it is possible that injection per se may influence the behavioral response. Rats injected with clomipramine hydrochloride (1, 10 or 50 mg/kg), as compared to animals injected with physiological saline as placebo, showed a dose-dependent decrease of the immobility time in the despair test and of the number of floor units explored in the open field in the reserpine test. However, when animals injected with placebo or clomipramine 50 mg/kg were compared with untreated intact controls, it was found that the immobility time in the despair test was higher in the placebo treated animals than in untreated intact controls. A difference was found between clomipramine-injected animals and untreated intact controls. In contrast, rats tested in the reserpine test, which is based on repeated drug injections, no difference was found between placebo-treated animals and untreated intact controls. These results indicate that stressful procedure of the experimental design may change the response of animals in behavioral tests. Studies with experimental models of depression, where stressful procedures are used, should include a control group of untreated intact animals. PMID- 11418281 TI - The prolactin response to sulpiride in major depression: the role of the D2 receptor in depression. AB - Multiple lines of investigations have implicated the role of the dopaminergic system in depression. The aim of the study was to characterise the Dopamine D2 receptor sensitivity status in depressed patients versus controls by means of a novel neuro-endocrine challenge test, the prolactin response to sulpiride. In this intervention, ten patients and ten age matched male volunteers were studied. The patients were diagnosed according to DSM-IV criteria, and Montgomery Asberg and Zung scales were done. There was no significant difference in baseline levels of prolactin between the depressed and control groups. Significantly higher prolactin levels after sulpiride challenge were however found in depressed patients than controls at all time points after sulpiride administration. This neuroendocrine challenge paradigm suggests that the prolactin response to sulpiride, a D2 receptor antagonist, is enhanced in depression, which suggests that this receptor might be supersensitive in depression compared to controls. This adds to the data implicating the dopaminergic system in the pathophysiology of depression, and suggests that dopaminergic mechanisms might be a target of therapeutic interest. PMID- 11418282 TI - The cAMP-dependent protein kinase substrate Rap1 in platelets from patients with obsessive compulsive disorder or schizophrenia. AB - Previous studies have reported that the cAMP-dependent protein kinase and one of its substrates, namely Rap1, are altered in patients with affective disorders. Abnormalities in the cAMP-dependent protein kinase have also been reported in platelets of patients with obsessive compulsive disorder and schizophrenia. However, it remains to be determined whether abnormalities in Rap1 are specifically related to affective disorders or may also be present in schizophrenia and obsessive compulsive disorder. Thus, we investigated Rap1 in platelets from 12 drug-free patients with obsessive compulsive disorder, ten drug free patients with schizophrenia, and 20 healthy subjects. While no difference was observed in the levels of Rap1 between groups, the phosphorylation state of Rap1 was significantly lower in patients with obsessive compulsive disorder than in schizophrenic patients and controls. These data further support the idea that abnormalities of cAMP signalling pathway could be associated, albeit in a somewhat different way, with several psychiatric disorders. PMID- 11418283 TI - Gender differences in the efficacy of fluoxetine and maprotiline in depressed patients: a double-blind trial of antidepressants with serotonergic or norepinephrinergic reuptake inhibition profile. AB - BACKGROUND: Depression has emerged as a contrastive area of gender differences in psychiatry, as epidemiological data has consistently shown depression is twice as common in women as men. The pharmacodynamic effect of antidepressants may also show gender differences, as suggested by reports of better response of young women to non-tricyclic antidepressants. METHODS: The antidepressive effect of an SSRI (fluoxetine) and a tetracyclic antidepressant with selective norepinephrine reuptake inhibitory effect (maprotiline) was compared in a 6-week, double-blind trial of 105 depressed patients. RESULTS: No significant difference was observed in the change of HAMD17 total score from baseline to week 6 between fluoxetine- and maprotiline-treated patients. A significant difference was observed in females (fluoxetine, -17.8; maprotiline, -13.9; P=0.017) between treatment groups, but not in males. Amongst females, the difference was significant in women aged <44 years (fluoxetine, -18.4; maprotiline, -12.9; P=0.023) but not > or =44 years. CONCLUSIONS: Females in their reproductive period are more responsive to SSRI (fluoxetine) than norepinephrinergic tetracyclic antidepressant (maprotiline) treatment. Normal cyclical ovulation, and estrogen release may have a clinically relevant pharmacodynamic interaction with serotonergic antidepressants. PMID- 11418284 TI - The acute effects of norharman on cocaine self-administration and sensorimotor function in male Wistar rats. AB - The purpose of the present study was to determine the acute effects of the beta carboline norharman on cocaine dependence. Male Wistar rats were allowed to self administer cocaine for 3 h for seven sessions. A single injection of norharman (0.2-20 mg/kg, i.p.) was given 30 min before the onset of the seventh session. It was shown that norharman decreased the cocaine intake in a U-shaped manner with the dose of 10 mg/kg having the most potent effect. These results indicate that several receptor mechanisms mediate the effects of norharman. In addition, 15 min following the administration of norharman on session 7, motor and sensory skill tests were performed. Six naive animals were tested with the dose, which has the most pronounced effect on cocaine self-administration intake, in order to examine whether the observed effects were due to norharman or due to the combination of norharman and cocaine. It was observed that norharman in both the naive and cocaine-exposed animals significantly increased the response time in the somato sensory orienting test. However, only in the naive animals a significant effect of norharman on the grasp reflex was observed. PMID- 11418285 TI - Heroin self-administration by means of 'chasing the dragon': pharmacodynamics and bioavailability of inhaled heroin. AB - In this controlled clinical study, the bioavailability and pharmacodynamics of inhaled heroin are evaluated and compared between 'chasing the dragon' and inhalation from a heating device, and at three dose levels, 25, 50 and 100 mg heroin, in two separate study phases. In study phase 1, no differences between the inhalation methods were detected on any of the physiological or behavioral measures, nor in bioavailability. Subjectively, the participants had a strong preference for the method of chasing, which was therefore used in study phase 2. In phase 2, heroin produced a dose-related increase in subjective drug-liking, body temperature and heart rate, and a clear, dose-related decline in reaction time. Linearly dose-related differences were found in the amount of total morphine in urine, amounting to an average of 45% of the parent heroin base received. Based on these findings, it is concluded that chasing is quite an effective route of heroin administration, producing rapid, dose-related subjective and objective effects and a sufficiently high and reproducible bioavailability. PMID- 11418286 TI - Clinical investigation of medicinal products in the treatment of epileptic disorders. PMID- 11418297 TI - Immunocytochemical detection of leukocyte-associated and apoptosis-related antigen expression in childhood brain tumors. AB - During systematic cell-surface antigen expression profile analyses of 76 primary childhood brain tumors [34 medulloblastomas (MED)/primitive neuroectodermal tumors (PNETs) and 42 astrocytomas (ASTR)], a library of monoclonal antibodies (MoABs) directed against various leukocyte-associated, lymphocyte cell-line differentiation antigens in childhood brain tumors was utilized. The antigens were detected employing an indirect, biotin-streptavidin conjugated alkaline phosphatase (AP) immunocytochemical technique. Major histocompatibility complex (MHC) class I restricted, tumor-associated antigen (TAA) specific, CD8(+) cytotoxic T lymphocytes (CTL) were identified in 58/76 (76.32%) brain tumors, and usually represented 1-10% of all cells, but in some cases 30-44% of the cells were CD8(+). CD4(+), MHC class II restricted helper lymphocytes were present in 65/76 (85.53%) brain tumors, and accounted for 1-10% of the observed cells. Macrophages were present in 74/76 (97.37%) brain tumors, and their number also represented 1-10% of all observed cells in the brain tumor frozen sections. Leukocyte common antigen (LCA) expression was detected in all 76 (100%) brain tumors studied. MoAB UJ 308 detected the presence of premyelocytes and mature granulocytes in 60/76 (78.95%) brain tumors. Natural killer (NK) cells were not defined in the observed brain tumors. The great majority of childhood glial tumors, particularly ASTRs express Fas (APO-1/CD95) receptor whereas normal cells in the central nervous system (CNS) do not. FasR is a transmembrane glycoprotein which belongs to the nerve growth factor/tumor necrosis factor (NGF/TNF) receptor superfamily. As part of our screening, the 42 childhood ASTRs were also investigated for expression of CD95. We detected strong expression (strong intensity of staining, number of stained cells 50-100%) of FasR, employing formalin fixed, paraffin-wax embedded tissue slides. Brain tumors and melanomas have been shown to produce their autocrine FasL, and are even capable of switching CD95-related signal transduction from the PCD pathway to a proliferative pathway. In view of our results, we conclude that: (1) the tumor infiltrating leukocytes in MEDs/PNETs and ASTRs represent a very diverse population and are present in a great majority of the cases studied; (2) the strong expression of FasR in ASTRs provides a manner in which T lymphocytes may exert their anti-tumor effects, but may also represent yet another way that tumors may evade the immune response; and (3) further observations of the expression of various antigens involved in juxtacrine, in situ growth control are necessary for the refinement of cellular immunotherapeutical approaches in the treatment of human malignancies. PMID- 11418298 TI - Treatment of refractory recurrent malignant glioma with adoptive cellular immunotherapy: a case report. AB - We report the successful treatment of a patient with recurrent malignant glioma with adoptive cellular immunotherapy. The patient is a young adult with recurrent progressive disease refractory to aggressive multi-modality therapy including repetitive surgical resection, radiation, radiosurgery and chemotherapy. He received multiple courses of local administration of autologous lymphokine activated killer (LAK) cells in combination with a low dose of interleukin-2 (IL 2) through an Ommaya reservoir-catheter system. The side-effects of this treatment were limited and manageable. The patient achieved a complete remission, as demonstrated by MRI and confirmed by glucose-positron emission tomography (PET) imaging 11 months after initiation of immune therapy. Twenty-six months later, the patient is still in remission with improving performance status. Adoptive cellular immunotherapy utilizing autologous LAK cells with low dose IL-2 appears to be a safe and effective therapy for a subset of patients with primary, recurrent or progressive malignant glioma following conventional therapy. PMID- 11418299 TI - Non-myeloablative allogeneic stem cell transplantation focusing on immunotherapy of life-threatening malignant and non-malignant diseases. AB - Allogeneic bone marrow transplantation (BMT) represents an important therapeutic tool for treatment of otherwise incurable malignant and non-malignant diseases. Until recently, myeloablative regimens were considered mandatory for eradication of all undesirable host-derived hematopoietic elements. Our preclinical and ongoing clinical studies indicated that much more effective eradication of host immunohematopoietic system cells could be achieved by adoptive allogeneic cell therapy with donor lymphocyte infusion (DLI) following BMT. Thus, eradication of blood cancer cells, especially in patients with CML can be frequently accomplished despite complete resistance of such tumor cells to maximally tolerated doses of chemoradiotherapy. Our cumulative experience suggested that graft versus leukemia (GVL) effects might be a useful tool for eradication of otherwise resistant tumor cells of host origin. The latter working hypothesis suggested that effective BMT procedures may be accomplished without lethal conditioning of the host, using new well tolerated non-myeloablative regimen, thus possibly minimizing immediate and late side effects related to myeloablative procedures considered until recently mandatory for conditioning of BMT recipients. Recent clinical data that will be presented suggests that safe non myeloablative stem cell transplantation (NST), with no major toxicity can replace the conventional BMT. Thus, NST may provide an option for cure for a large spectrum of clinical indications in children and elderly individuals without lower or upper age limit, while minimizing procedure-related toxicity and mortality. PMID- 11418300 TI - Adoptive cellular immunotherapy for the treatment of malignant gliomas. AB - The median survival for adults with recurrent primary malignant gliomas is 56 weeks following surgery, radiation, and chemotherapy. Generally, reoperation can extend the median survival an additional 26-32 weeks. We have developed an aggressive treatment program that utilizes low doses of interleukin-2 (IL-2) combined with ex vivo activated killer cells (LAK) infused via an indwelling catheter placed into the surgical resection cavity. Autologous leukocytes were collected during a standard 3-4 h, outpatient leukapheresis procedure, then activated ex vivo for 4-5 days with high doses of IL-2. The treatment protocol consisted of two 2-week cycles of therapy over a 6-week period. Patients with stable disease or objective response on follow-up MRI scans were retreated at 3 month intervals. Acute and cumulative IL-2-related toxicities were observed, but limited, and included fever, headache and transient neurologic irritation. Corticosteroid levels and usage were strictly controlled during immunotherapy, although higher doses were used intermittently to mitigate toxicity. Biologic changes included lymphocytic infiltration, regional eosinophilia, tumor necrosis, and the localized production of IL-2, IFN-gamma and IL-12, demonstrated by in situ hybridization and immunohistochemistry. SUMMARY: IL-2 plus autogeneic LAK cells can be safely administered intracavitary to treat high grade primary brain tumors with limited toxicity within the central nervous system. Six out of 28 patients had long-term survival of greater than 2 years post-reoperation plus immunotherapy with 2 patients alive over 8 years. The presence of a marked regional eosinophilia appeared to correlate with increased survival and may be predictive of a biologic and therapeutic response. Regional adoptive immune therapy was well tolerated and should be considered an option for patients with high-grade tumors refractive to standard therapeutic approaches. PMID- 11418301 TI - Vector-driven hyperexpression of a triad of costimulatory molecules confers enhanced T-cell stimulatory capacity to DC precursors. AB - Activation of T cells requires at least two signals: signal 1, via the T-cell receptor, and signal 2, in which a costimulatory molecule on the antigen presenting cell (APC) interacts with a ligand on the T cell. Dendritic cells (DCs) are the most potent APCs in part due to their expression of costimulatory molecules. DCs, however, constitute only a minor percentage of APCs in the body, and the in vitro preparation of DCs is both costly and time consuming. The studies reported here demonstrate that one can utilize other APCs, such as bone marrow progenitor cells (BMPCs) and make them markedly more effective as APCs; this was accomplished by their infection with recombinant poxviruses (either the replication-defective avipox or vaccinia), which contain transgenes for a triad of costimulatory molecules (B7-1, ICAM-1 and LFA-3, designated TRICOM). APCs infected with TRICOM vectors are shown to significantly enhance the activation of both naive and effector CD4(+) and CD8(+) T-cell populations. The use of TRICOM vectors in vaccine strategies is discussed. PMID- 11418302 TI - Susceptibility of B-cell lymphoma to human antibodies encoded by the V4-34 gene. AB - Our previous studies have shown that mAbs derived from the human V4-34 gene bind and kill human B-lymphocytes via membrane disruption. This study demonstrates the cytotoxicity of two V4-34 encoded mAbs, 216 and Z2D2, towards human B-cell lymphoma. In vitro, 216 and Z2D2 are cytotoxic to a variety of B-cell lymphomas obtained from patient biopsies. In vivo, increased survival was observed with both mAbs in a lymphoma model developed in scid mice with human B-cell line Nalm 6. Studies in mice show that these mAbs are well tolerated with minimum side effects. Since 216 and Z2D2 show increased toxicity towards cycling cells, V4-34 mAb-based therapy can be additive with drugs that block cell-cycle progression. Stem cells that are V4-34 mAb ligand negative would not be depleted. Together, these studies recommend an evaluation of the two completely human mAbs in a phase I trial for B-cell lymphoma. PMID- 11418303 TI - IgA antibodies for cancer therapy. AB - Antibody-based therapy is a new treatment option for selected tumor patients. Today, human IgG(1) is the most widely used isotype, because it effectively activates human complement, recruits NK cells for ADCC, and has an extended plasma half life. In recent work, however, neutrophils--the most populous cytotoxic cells in humans--were more effectively recruited by human IgA than by IgG antibodies. IgA antibodies may have the additional advantages of forming natural dimers with improved signaling capacity on tumor cells, and being actively transported into mucosal secretions with the potential for improved targeting of certain carcinomas from the luminal surface. PMID- 11418304 TI - Specifically targeting the CD22 receptor of human B-cell lymphomas with RNA damaging agents. AB - Targeting CD22 on human B-cells with a monoclonal antibody conjugated to a cytotoxic RNAse causes potent and specific killing of the lymphoma cells in vitro. This translates to anti-tumor effects in human lymphoma models in SCID mice. RNA damage caused by RNAses could be an important alternative to standard DNA damaging chemotherapeutics. Moreover, targeted RNAses may overcome problems of toxicity and immunogenicity associated with plant or bacterial toxin containing immunotoxins. PMID- 11418305 TI - Molecular targeting of malignant gliomas with novel multiply-mutated interleukin 13-based cytotoxins. AB - A vast majority of high-grade gliomas over-express a receptor for interleukin 13 (IL13). This glioma-associated receptor for IL13 is interleukin 4 (IL4) independent. This is in contrast to the physiological and IL4-shared receptor for the IL13, IL13/4 receptor, which is found on many normal organs. IL13-based Pseudomonas exotoxin (PE)-containing cytotoxic fusion proteins have been shown to be very potent anti-glioma agents. However, native IL13-based cytotoxins interact with both forms of the IL13 receptor. Therefore, mutations in IL13 were made in order to diminish/eliminate IL13's interaction with the shared IL13/4 receptor of normal tissue. These mutations encompassed amino acids located on alpha-helix A and C of IL13. We have engineered double or triple mutants of IL13 linked to various forms of PE. We found that these mutations could be successfully incorporated into IL13 without the loss of the protein's ability to selectively deliver the toxin to glioma cells while reducing their toxicity. PMID- 11418306 TI - The use of Shiga-like toxin 1 in cancer therapy. AB - The ribosome-inactivating protein, Shiga-like toxin-1 (SLT-1, SLT-I, Verotoxin 1, VT1) targets cells that express the glycolipid globotriaosylceramide (CD77) on their surface. The frequent occurrence of SLT-1 receptors on tumor cells derived from patients with hematological cancers (follicular lymphoma, multiple myeloma, chronic lymphocytic leukemia) and their absence on human CD34(+) hematopoietic stem cells suggest the ex vivo use of Shiga-like toxin-1 in purging CD77(+) tumor cells from autologous stem cell transplants. SLT-1 receptors are also commonly expressed on breast cancer, ovarian cancer and astrocytoma cells. In particular, the sensitivity of astrocytoma cell lines to this toxin provides an opportunity for using SLT-1 in vivo in the context of treating patients afflicted by this common form of brain tumor. Finally, the known structural features of SLT-1 allow one to contemplate altering its receptor specificity in an effort to target CD77( ) tumor cell populations. PMID- 11418307 TI - Anti-idiotypic antibody and recombinant antigen vaccines in colorectal cancer patients. AB - The colorectal carcinoma (CRC)-associated GA733 antigen (also known as CO17-1A, KS1-4, KSA or EpCAM) has been the target of a phase II/III randomized trial of passive immunotherapy with monoclonal antibody CO17-1A and phase I active immunotherapy trials with polyclonal anti-idiotypic antibodies mimicking the CO17 1A or GA733 epitope on the antigen. The CO17-1A antigen was molecularly cloned and the extracellular domain expressed in baculovirus (BV) GA733-2E. Whereas, anti-idiotypic antibody mimics a single epitope on the antigen, BV GA733-2E expresses multiple potentially immunogenic epitopes. In animals, the immunogenicity of BV GA733-2E in aluminum hydroxide was superior to that of anti idiotype in the same adjuvant. Here, we compared the immunogenicity of anti idiotypic antibody and GA733-2E antigen in CRC patients. These studies indicate that the antigen is superior to the anti-idiotype antibody in inducing humoral and cellular immunity in CRC patients. PMID- 11418308 TI - Short-term autologous tumor cell lines for the active specific immunotherapy of patients with metastatic melanoma. AB - We established short-term cell lines for 108/170 (64%) patients with metastatic melanoma. Tumor cell numbers were expanded to 10(8), then cells were irradiated, aliquoted, and cryopreserved for clinical use. Vaccines have been used to treat 69 patients with clinical follow up for 33 who had measurable metastatic disease at the time vaccine therapy was initiated (METS), and 33 who had no evidence of disease (NED) at the time of vaccine therapy following surgical resection of metastases. The protocol called for a baseline test of delayed tumor hypersensitivity (DTH), three weekly injections, a repeat of the DTH test, then monthly injections for an additional 5 months. Objective tumor responses were noted in 3/26 (12%) patients who received a minimum of three vaccinations, one complete, and two partial, with survivals of 36, 46+, and 78+ months. Only 6/64 (9.4%) had a positive DTH (>10 mm) at baseline, including three METS, all of whom progressed within 4 months and died within a year, and three who are still NED after more than 5 years. Conversion of DTH from negative to positive was documented in 18/44 (41%) patients who were tested at week 0 and 4. At a median follow up of greater than 5 years, the median overall survival (OS) was 40 months for "NED" with a 5-year survival rate of 39%, and 8.6 months with a 5-year survival rate of 10% for "METS" The 18 patients who had conversion of their DTH had a median event-free survival (EFS) of 15.8 months and 5-year EFS of 32% compared to 4.2 months and 9% for the 26 non-converters (P=0.012, two-tailed, log rank test). Among patients who were NED when treatment started, the 12 patients whose DTH converted had a median overall survival of 61.4 months with 5-year survival of 63% compared to 9.7 months and 0% for the 13 non-converters (P=0.0026). This treatment approach is feasible, produces minimal toxicity, and is associated with long-term survival in a significant subset of patients. PMID- 11418309 TI - In vivo transfection and/or cross-priming of dendritic cells following DNA and adenoviral immunizations for immunotherapy of cancer--changes in peripheral mononuclear subsets and intracellular IL-4 and IFN-gamma lymphokine profile. AB - In order to provoke an immune response, a tumor vaccine should not only maximize antigen-specific signals, but should also provide the necessary "co-stimulatory" environment. One approach is to genetically manipulate tumor cells to either secrete lymphokines (GM-CSF, IL-12, IL-15) or express membrane bound molecules (CD80, CD86). Furthermore, patient dendritic cells can be loaded with tumor associated antigens or peptides derived from them and used for immunotherapy. Genetic modification of dendritic cells can also lead to presentation of tumor associated antigens. Transfection of dendritic cells with DNA encoding for such antigens can be done in vitro, but transfection efficiency has been uniformly low. Alternatively, dendritic cells can also be modulated directly in vivo either by "naked" DNA immunization or by injecting replication-deficient viral vectors that carry the tumor specific DNA. Naked DNA immunization offers several potential advantages over viral mediated transduction. Among these are the inexpensive production and the inherent safety of plasmid vectors, as well as the lack of immune responses against the carrier. The use of viral vectors enhances the immunogenicity of the vaccine due to the adjuvant properties of some of the viral products. Recent studies have suggested that the best strategy for achieving an intense immune response may be priming with naked DNA followed by boosting with a viral vector. We have successfully completed a phase I and phase II clinical trials on immunotherapy of prostate cancer using naked DNA and adenoviral immunizations against the prostate-specific membrane antigen (PSMA) and phase I clinical trial on colorectal cancer using naked DNA immunization against the carcinoembryonic antigen (CEA). The vaccination was tolerated well and no side effects have been observed so far. The therapy has proven to be effective in a number of patients treated solely by immunizations. The success of the treatment clearly depends on the stage of the disease proving to be most efficient in patients with minimal disease or no metastases. A panel of changes in the phenotype of peripheral blood lymphocytes and the expression of intra-T cell lymphokines seems to correlate with clinical improvement. PMID- 11418310 TI - A rapid method to identify cytotoxic T-lymphocyte peptide epitopes from HLA-A2 (+) donors. AB - It would be useful to develop a method to rapidly identify peptide epitopes for vaccine development. We present an algorithm that can predict sequences that have a high binding activity for HLA-A2. These sequences were able to induce specific cytolytic cells from human peripheral blood lymphocytes (PBMC). A computer assisted algorithm was constructed to predict binding activity for HLA-A2, according to anchoring amino acid combinations. The human papillomavirus (HPV) type 18 E7 oncoprotein was used to test the algorithm. Peptides predicted to bind were synthesized and binding activity was determined by using the T2 cell assay. T2 cells pulsed with HPV-18 peptides were incubated with PBMC. Cytotoxicity assays were performed. From 110 possible sequences, four peptides were found to have a high binding activity. One of these peptides was able to induce significant lysis. Using this selection process only 3.6% of the total number of possible sequences was synthesized to identify an immunogenic peptide. Our algorithm with the T2 binding assay allows a rapid method to detect peptide epitopes. PMID- 11418311 TI - In vitro and preclinical targeted alpha therapy for melanoma, breast, prostate and colorectal cancers. AB - Targeted alpha therapy (TAT) can inhibit the growth of micrometastases by selectively killing isolated and preangiogenic clusters of cancer cells. The alpha emitting radioisotopes Tb-149 and Bi-213 were chelated to cancer specific monoclonal antibodies to form alpha-immunoconjugates (AIC) against melanoma, leukaemia, prostate and colorectal cancer, and to the plasminogen activator inhibitor type-2 (PAI2) to form alpha-PAI2 (API) against breast and prostate cancer. These conjugates were found to be highly stable, specific and cytotoxic in vitro. Melanoma and breast cancer tumour growth was observed in nude mouse models for untreated controls and non-specific AIC/API at 2 days post subcutaneous inoculation of cancer cells. Complete inhibition of melanoma and breast cancer growth was found for local injections of AIC and API, respectively. Intra-lesional TAT of established melanoma showed that all melanomas regressed with 100 microCi injections of AIC. These results point to the potential application of local and systemic TAT in the management of metastatic cancer. PMID- 11418312 TI - Localization of pancreatic cancer with radiolabeled monoclonal antibody PAM4. AB - Experimental animal studies were performed with (111)In-labeled PAM4 anti-MUC1 antibody along with (111)In-labeled control antibody. Tumor uptake of radiolabeled PAM4 was significantly higher than for the control antibody at all time points. When normalized to a blood dose of 1500 cGy as an estimate of myelotoxicity, (90)Y-labeled PAM4 would provide 5344 cGy to the tumor, whereas an equitoxic dose of (90)Y-labeled control antibody would provide only 862 cGy to the tumor. In addition to the animal studies, five patients with proven pancreatic cancer were administered either (131)I-PAM4 IgG (n=2) or 99mTc-PAM4 Fab' (n=3). Tumor targeting was observed in four out of five patients. By immunohistochemistry, PAM4 was non-reactive with tumor from the one patient not targeted. Dosimetry from the patients given (131)I-PAM4 predicted that tumors would receive 10-20 cGy/mCi with tumor/red marrow dose ratios ranging from 3 to 10. Based upon these results, we have established a phase-I (111)In-labeled PAM4 imaging and (90)Y-labeled PAM4 therapy trial. PMID- 11418313 TI - Antiangiogenic agents and their promising potential in combined therapy. AB - One of the most promising strategies for treating cancer is the addition of antiangiogenic therapy to therapeutic regimens. Angiogenesis, or the growth of new blood vessels from preexisting vessels, is essential both for the growth of a primary tumor and for successful metastasis. As a result of intense research in this field, a number of antiangiogenic agents have been identified and have demonstrated varying degrees of success in inhibiting the growth of solid tumors and metastases in preclinical and clinical studies. The real potential of antiangiogenic agents for cancer therapy resides in strategic combinations with each other, with chemotherapy, with radiation, and with tumor-targeting agents, such as radioimmunotherapy. Along with this new opportunity to develop synergistic therapy comes the challenging complexities of the physiologic systems regulating angiogenesis. These multifaceted systems could intimidate investigators seeking to take advantage of the potential synergy in combined cancer therapy. To aid in these efforts, this overview of key antiangiogenic agent mechanisms, combination strategies and initial studies of the potential synergy with chemotherapy, radiation and radioimmunotherapy is presented. PMID- 11418314 TI - Successful therapy of a human lung cancer xenograft using MAb RS7 labeled with residualizing radioiodine. AB - We have recently reported that a radioiodinated, DTPA-appended peptide, designated IMP-R1, is a residualizing iodine label that overcomes many of the limitations that have impeded the development of residualizing iodine for clinical use. In this study the potential of 131I-IMP-R1-RS7, an internalizing anti-EGP-1 monoclonal antibody, was evaluated by performing preclinical therapy studies in nude mice bearing Calu-3 human non-small cell carcinoma of the lung xenografis. Elimination of 6 of 9 established tumors (mean tumor volume=0.3 cm(3)) was observed using a single dose of 350 microCi/mouse of 131I-IMP-R1-RS7, with all animals tolerating the dose. At the same dose and specific activity of 131I-RS7, labeled using the conventional chloramine-T method, there were four deaths, and one complete remission in nine treated mice. At the maximum tolerated dose of conventionally 131I-labeled RS7, 275 microCi, mean stable disease for approximately 5 weeks was observed, with no complete responses. Specificity of the therapeutic effect was shown in an isotype-matched control experiment, where 131I-IMP-R1-RS7 was markedly more effective than the (131)I-IMP-R1-labeled control antibody. These studies demonstrate that (131)I-IMP-R1-RS7 provides a therapeutic advantage in comparison to conventional 131I-labeled RS7, as predicted by the increased tumor accretion observed previously in targeting studies. A direct comparison of the maximum tolerated doses of (131)I-IMP-R1-RS7 (350 microCi) and 90Y-DOTA-RS7 (105 microCi) was performed in this tumor model using large established tumors (mean tumor volume=0.85 cm(3)). Anti-tumor efficacy and toxicity of the two treatments were comparable. PMID- 11418315 TI - Biodistribution and dosimetry results from a phase III prospectively randomized controlled trial of Zevalin radioimmunotherapy for low-grade, follicular, or transformed B-cell non-Hodgkin's lymphoma. AB - Radiation dosimetry studies were performed in patients with non-Hodgkin's lymphoma (NHL) treated with 90Y Zevalin (90yttrium ibritumomab tiuxetan, IDEC Y2B8) on a Phase III open-label prospectively randomized multicenter trial. The trial was designed to evaluate the efficacy and safety of 90Y Zevalin radioimmunotherapy compared to rituximab (Rituxan, MabThera) immunotherapy for patients with relapsed or refractory low-grade, follicular, or transformed NHL. An important secondary objective was to determine if radiation dosimetry prior to 90Y Zevalin administration is required for safe treatment in this patient population. METHODS: Patients randomized into the Zevalin arm were given a tracer dose of 5 mCi (185 MBq) (111)In Zevalin (111indium ibritumomab tiuxetan) on Day 0, evaluated with dosimetry, and then administered a therapeutic dose of 0.4 mCi/kg (15 MBq/kg) 90Y Zevalin on Day 7. Both Zevalin doses were preceded by an infusion of 250 mg/m(2) rituximab to clear peripheral B-cells and improve Zevalin biodistribution. Following administration of (111)In Zevalin, serial anterior and posterior whole-body scans were acquired and blood samples were obtained. Residence times for 90Y were estimated for major organs, and the MIRDOSE3 computer software program was used to calculate organ-specific and total body radiation absorbed dose. Patients randomized into the rituximab arm received a standard course of rituximab immunotherapy (375 mg/m(2) weekly x 4). RESULTS: In a prospectively defined 90 patient interim analysis, the overall response rate was 80% for Zevalin vs. 44% for rituximab. For all patients with Zevalin dosimetry data (N=72), radiation absorbed doses were estimated to be below the protocol-defined upper limits of 300 cGy to red marrow and 2000 cGy to normal organs. The median estimated radiation absorbed doses were 71 cGy to red marrow (range: 18-221 cGy), 216 cGy to lungs (94-457 cGy), 532 cGy to liver (range: 234 1856 cGy), 848 cGy to spleen (range: 76-1902 cGy), 15 cGy to kidneys (0.27-76 cGy) and 1484 cGy to tumor (range: 61-24274 cGy). Toxicity was primarily hematologic, transient, and reversible. The severity of hematologic nadir did not correlate with estimates of effective half-life (half-life) or residence time of 90Y in blood, or radiation absorbed dose to the red marrow or total body. CONCLUSION: 90Y Zevalin administered to NHL patients at non-myeloablative maximum tolerated doses delivers acceptable radiation absorbed doses to uninvolved organs. Lack of correlation between dosimetric or pharmacokinetic parameters and the severity of hematologic nadir suggest that hematologic toxicity is more dependent on bone marrow reserve in this heavily pre-treated population. Based on these findings, it is safe to administer 90Y Zevalin in this defined patient population without pre-treatment (111)In-based radiation dosimetry. PMID- 11418316 TI - The role of radiolabeled antibodies in the treatment of non-Hodgkin's lymphoma: the coming of age of radioimmunotherapy. AB - This review summarizes the current clinical status of radioimmunotherapy (RAIT) in the treatment of patients with non-Hodgkin's lymphoma (NHL), as a prototype of the advances of RAIT in the management of cancer. Four radiolabeled antibody products are progressing towards commercialization for the RAIT of NHL: 131I tositumomab (Bexxar), 90Y-ibritumomab tiuxetan, 90Y-epratuzumab (hLL2), and 131I Lym-1. All except epratuzumab are murine monoclonal antibodies (Mabs) labeled with an isotope, except that ibritumomab (Zevalin) adds chimeric rituximab to the product, whereas epratuzumab is solely a humanized Mab. Bexxar and Zevalin target CD20, epratuzumab binds to CD22, and Lym-1 reacts with HLA-DR. Clinical studies have shown that all four antibody products can be safe and efficacious. Bexxar has been shown to induce responses that are relatively better than the prior chemotherapy, and has also been shown to be effective in combination with chemotherapy as a frontline therapy of low-grade and transformed NHL. However, since it is a fully murine Mab, it did show a approximately 60% HAMA rate in untreated patients. Zevalin has been found to be more effective than rituximab, its naked chimeric Mab counterpart, as well as in chemotherapy-relapsed low-grade NHL patients. Both radiolabeled epratuzumab and Lym-1 have shown efficacy in patients who have failed chemotherapy, either with low-grade or aggressive forms of NHL. It appears that Bexxar and Zevalin will be the first two radiolabeled antibodies that may be available for widespread use in the U.S., and will mark the final introduction of RAIT as an approved cancer treatment modality. Future studies will help define the role of these RAIT products in the management of NHL, especially as part of a multimodal therapy of this disease. PMID- 11418317 TI - Role of radiation dosimetry in radioimmunotherapy planning and treatment dosing. AB - Cancer-seeking antibodies (Abs) carrying radionuclides can be powerful drugs for delivering radiotherapy to cancer. As with all radiotherapy, undesired radiation dose to critical organs is the limiting factor. It has been proposed that optimization of radioimmunotherapy (RIT), that is, maximization of therapeutic efficacy and minimization of normal tissue toxicity, depends on a foreknowledge of the radiation dose distributions to be expected. The necessary data can be acquired by established tracer techniques, in individual patients, using quantitative radionuclide imaging. Object-oriented software systems for estimating internal emitter radiation doses to the tissues of individual patients (patient-specific radiation dosimetry), using computer modules, are available for RIT, as well as for other radionuclide therapies. There is general agreement that radiation dosimetry (radiation absorbed dose distribution, cGy) should be utilized to establish the safety of RIT with a specific radiolabeled Ab in the early stages (i.e. phase I or II) of drug evaluation. However, it is less well established that radiation dose should be used to determine the radionuclide dose (amount of radioactivity, GBq) to be administered to a specific patient (i.e. radiation dose-based therapy). Although treatment planning for individual patients based upon tracer radiation dosimetry is an attractive concept and opportunity, particularly for multimodality RIT with intent to cure, practical considerations may dictate simpler solutions under some circumstances. PMID- 11418318 TI - Humoral mechanisms in prostate cancer: A role for FSH. AB - The natural history of prostate cancer has long been related to the male hormone testosterone, and treatment has focused on depletion of this androgen to slow or prevent growth of prostate cancer tissue. It has become clear recently, however, that more than androgens influence the progression of prostate cancer, with recent interest focusing on the gonadotropin, follicle-stimulating hormone (FSH). Research of the last decade has found that FSH is produced in and FSH receptors are expressed in the prostate. Investigators have found as well that production of FSH is altered in prostate cancer: FSH levels are increased and receptor production raised in the cancerous prostate. It also has been shown that there are endogenous compounds such as prostatic inhibin peptin that can modulate FSH levels. All of these findings are outlined in this paper, and suggest that FSH may affect the pathogenesis and progression of prostate cancer and that altering FSH production may prove to be an active therapeutic maneuver. PMID- 11418319 TI - Surveillance for stage I testicular seminoma. a review. AB - There is good evidence that patients with stage I seminoma can be managed equally well after orchidectomy with surveillance and adjuvant retroperitoneal radiation therapy. There is considerable reluctance amongst many physicians to accept surveillance as a management option in stage I seminoma and this is largely based on the excellent results achieved with adjuvant retroperiteonal radiation for many years. However, patients with stage I seminoma have a long life span and it is possible that the long-term sequelae of radiation treatment could have a negative impact on quality of life and longevity. It is of utmost importance to continue the study of the long-term effects of all current treatment approaches, in particular the risk of induction of second malignancies. However, the psychosocial impact on patients of surveillance and other management strategies must also be assessed. Stage I testicular seminoma is highly curable with currently available management approaches and the current challenge for clinicians is to maintain these excellent results while minimizing toxicity and individualising treatment to the specific social, economic and emotional circumstances of each patient. Surveillance should be one of the management options offered to patients with stage I seminoma. PMID- 11418320 TI - Phase II evaluation of suramin in advanced renal cell carcinoma. A Southwest Oncology Group study. AB - Twenty-two eligible patients with advanced renal carcinoma were treated with suramin utilizing a fixed dose regimen. Therapy was reasonably well tolerated with 3 of 22 patients (14%) developing grade 4 toxicity and 11 of 22 patients (50%) having a maximum toxicity of grade 3. There were no responders; median survival was 10 months. Suramin is not an active agent in advanced renal carcinoma. PMID- 11418321 TI - Telomerase activity in bilharzial bladder cancer. Prognostic implications. AB - Background: Bladder cancer is a common malignancy in Egypt and other developing countries in which infection with Schistosoma haematobium is prevalent. Bladder cancer caused by bilharziasis has different clinical and biological characters than that observed in the western world. In this study, we used the TRAP technique to estimate telomerase activity in bilharzial bladder cancer specimens and we correlated the findings with other clinical and pathological findings. Patients and methods: Bladder cancer specimens were obtained from 57 patients who underwent radical cystectomy and pathological diagnosis was obtained in all patients. Tissue samples were frozen in liquid nitrogen and stored at -80 degrees C. Telomerase activity by PCR-ELISA technique was measured using TRAP technique. Results: Our patient group included 45 males and 12 females with a median age of 49 years. The majority of our patients (35/57) have squamous histology and they have proven bilharzial history shown in the pathology specimens. Stage P3b was encountered in 29/57 patients whereas thirty-five patients have grade II tumors. The majority of our patients (41/57) were negative for pelvic nodes metastases. Telomerase activity was detected in 27/57 patients (47.4%). The mean level of telomerase was 0.85+/-0.77 in positive patients and 0.029+/-0.025 in negative patients. The expression of telomerase and its mean level in patients above age of 60, in males and in those with squamous pathology, higher grade of tumors or positive node was higher than those without but the difference did not reach statistical significance (P>0.05). Alternatively, expression was significantly higher in those with stages (P1-P3a) compared with P3b-P4a disease stages (66.6% vs. 37.1, P=0.03). Conclusion: Telomerase activity is increased in bilharzial bladder cancer although to a lesser degree than that reported for TCC in the western world, which could be explained, by different biological behavior or different assay methods. Further larger studies with more number of patients are still needed to determine its potential value for early detection and possible use as a therapeutic target. PMID- 11418322 TI - Growth inhibitory effect of p21 and p53 containing adenoviruses on transitional cell carcinoma cell lines in vitro and in vivo. AB - Altered p53 expression has been demonstrated in the majority of advanced transitional cell carcinoma (TCC) of the bladder tumors. The objective of this investigation was to examine the effect of the introduction of a p53 or p21((WAF1/CIP1)) adenovirus on the proliferation and apoptosis of various human TCC cell lines in vitro and in vivo. Proliferation was measured by 3H-thymidine incorporation. Apoptosis was measured by DNA fragmentation and bax expression. We also examined the effect of ex vivo introduction of the p21((WAF1/CIP1)) or the p53 gene on growth of the T24 TCC cells and UMUC-3 TCC cells introduced subcutaneously into athymic nude mice. We found that although the effect of the p21-adenovirus on the proliferation of various TCC lines varied with each individual cell line, there was a substantial growth inhibition observed (greater than 80% growth inhibition) in seven of the eight TCC cell lines at the highest viral dosage. In contrast, after 24 h, the highest dosage of the p53-adenovirus produced only a heterogeneous decrease in proliferation compared to the highest dose of the p21((WAF1/CIP1))-adenovirus (40-90%). In ex vivo experiments, no tumors were found in nude mice injected subcutaneously with either TCC cell line exposed in vitro to the AdSCMV-p21((WAF1/CIP1)) or AdSCMV-p53 viruses before three weeks. There was a threefold decrease in tumor square area at week 5 in the Ad5CMV-p21((WAF1/CIP1)) or Ad5CMV-p53 TCC cells injected mice (p<0.001, p<0.009) compared to either mock or Ad5CMVLacZ TCC bladder tumor cells. These data suggest that significant portion of the effect of altered p53 on TCC phenotype may be mediated through the p21((WAF1/CIP1)) pathway. Thus, the restoration of p21((WAF1/CIP1)) function in this tumor system may be a beneficial therapeutic strategy. PMID- 11418323 TI - Soluble Fas and Fas-ligand in bladder cancer in vitro and in vivo. AB - Circulating soluble Fas (sFas) and expression of Fas-ligand on cancer cells are mechanisms of immune escape. The aim of the present study was to investigate expression and production of Fas and Fas-ligand on bladder cancer cell lines of different grade as a basic mechanism of their secretion in vivo. sFas and sFas ligand serum levels of patients with different stage of bladder cancer were examined to determine the possible clinical use of these molecules as tumor markers. Bladder cancer cell lines RT4 (G1), RT112 (G1), T24 (G3) and SUP (G4) were analyzed by flowcytometry for Fas and Fas-ligand expression. To determine if the Fas-ligand gene is transcribed in these bladder cancer cell lines, RT-PCR was performed on mRNA extracted from these cell lines. Production of sFas and sFas ligand was examined in cell culture supernatants of the cancer cells as well as in the serum of 62 patients with bladder cancer by a specific ELISA test. We demonstrate that Fas is expressed in similar levels on all human bladder carcinoma cell lines. In T24 (G3) and SUP (G4) cell lines we were able to detect the Fas-ligand protein, whereas no Fas-ligand protein could be found in RT4 and RT112 (G1) cells. Fas-ligand mRNA was expressed in all bladder cancer cell lines. Furthermore, all bladder cancer cell lines produce sFas but no sFas-ligand in spite of mRNA expression. The range of sFas levels in the serum of all patients with bladder cancer was large and did not show a correlation to the histopathological stage of bladder cancer. Although there is in vitro evidence that sFas and Fas-ligand play a role in bladder cancer, no correlation between the sFas and s Fas-ligand serum levels and the histopathological stage of bladder cancer could be found. Therefore, serum sFas and sFas-ligand have to date limited clinical relevance. PMID- 11418325 TI - Erratum. PMID- 11418324 TI - Combined modality staging for clinically localized adenocarcinoma of the prostate as the basis for patient selection for randomized trials of neoadjuvant chemotherapy. PMID- 11418326 TI - Impact of transcription factors AP-1 and NF-kappaB on the outcome of experimental Staphylococcus aureus arthritis and sepsis. AB - Staphylococcus aureus infection is, despite adequate antibiotic treatment, a disease characterized by high mortality. The bacterium triggers an exaggerated immune response in the host, which on the one hand acts as an efficient defense, but on the other hand gives rise to tissue damage. In this study we have modulated the hosts response to S. aureus by inhibition of nuclear factor kappaB (NF-kappaB) and activator protein-1 (AP-1)-triggered release of pro-inflammatory cytokines and tissue-destructive proteins, respectively. Mice were administered with antisense oligonucleotides (ODN) to the p65 subunit of NF-kappaB and/or a double-stranded oligonucleotide (mCoAP-1) with homology to the murine AP-1 binding site of collagenase IV gene (metalloproteinase-9; MMP-9), solely or in combination with antibiotics. In mice systemically treated with antisense ODN to NF-kappaB p65 alone, the bacterial burden in the kidneys was significantly increased (P = 0.04) The same tendency was seen when mCoAP-1 was administered either alone or in combination with antibiotics. We also found significantly (P = 0.04) elevated levels of IL-6 in p65 antisense treated mice. Surprisingly, this p65 antisense therapy approach, which has turned out to be highly efficient in amelioration of aseptic arthritis and colitis, failed to change the clinical course of either septic arthritis or sepsis. We suggest that interaction with transcription factors leads to increased bacterial burden in vivo, abrogating the potential benefits of the anti-inflammatory properties exerted by these compounds. PMID- 11418327 TI - Two-dimensional electrophoresis and characterization of antigens from Paracoccidioides brasiliensis. AB - Paracoccidioides brasiliensis is a fungal pathogen of humans. To identify antigens from P. brasiliensis we fractionated a crude preparation of proteins from the fungus and detected the IgG reactive proteins by immunoblot assays of yeast cellular extracts with sera of patients with paracoccidioidomycosis (PCM). We identified and characterized six new antigens by amino acid sequencing and homology search analyses with other proteins deposited in a database. The newly characterized antigens were highly homologous to catalase, fructose-1,6 biphosphate aldolase (aldolase), glyceraldehyde-3-phosphate dehydrogenase, malate dehydrogenase and triosephosphate isomerase from several sources. The characterized antigens presented preferential synthesis in yeast cells, the host fungus phase. PMID- 11418328 TI - Pathogenicity islands: the tip of the iceberg. AB - Pathogenicity islands represent distinct genetic elements encoding virulence factors of pathogenic bacteria. Pathogenicity islands belong to the class of genomic islands, which are common genetic elements sharing a set of unifying features. Genomic islands have been acquired by horizontal gene transfer. In recent years many different genomic islands have been discovered in a variety of pathogenic as well as non-pathogenic bacteria. Because they promote genetic variability, genomic islands play an important role in microbial evolution. PMID- 11418329 TI - Salmonella pathogenicity islands encoding type III secretion systems. AB - Salmonella enterica harbours two Salmonella pathogenicity islands (SPIs) each encoding a type III secretion system for virulence proteins. SPI1 is required for invasion, while systemic infections and intracellular accumulation of Salmonella are dependent on SPI2 function. This review will describe and compare the genetic organisation, evolution, regulation and molecular functions of SPI1 and SPI2. PMID- 11418330 TI - The Yersinia high-pathogenicity island: an iron-uptake island. AB - Highly pathogenic Yersinia carry a pathogenicity island termed high-pathogenicity island (HPI). The Yersinia HPI comprises genes involved in the synthesis of the siderophore yersiniabactin and can thus be regarded as an iron-uptake island. A unique characteristic of the HPI is its wide distribution among different enterobacteria such as Escherichia coli, Klebsiella, Citrobacter and Salmonella. Other types of iron-uptake systems are also carried by different pathogenicity islands in enterobacteria. PMID- 11418331 TI - Pathogenicity islands and virulence evolution in Listeria. AB - As in other bacterial pathogens, the virulence determinants of Listeria species are clustered in genomic islands scattered along the chromosome. This review summarizes current knowledge about the structure, distribution and role in pathogenesis of Listeria virulence loci. Hypotheses about the mode of acquisition and evolution of these loci in this group of Gram-positive bacteria are presented and discussed. PMID- 11418332 TI - Pathogenicity and resistance islands of staphylococci. AB - Variable genetic elements including plasmids, transposons and prophages are involved in pathogenesis and antibiotic resistance, and are an important component of the staphylococcal genome. This review covers a set of newly described variable chromosomal elements, pathogenicity and resistance islands, carrying superantigen and resistance genes, especially toxic shock and methicillin resistance, respectively. PMID- 11418334 TI - Web alert. Biotic interactions. PMID- 11418336 TI - Putting knowledge of plant disease resistance genes to work. AB - Plant disease resistance genes trigger defence mechanisms upon recognition of pathogen compatibility factors, which are encoded by avirulence genes. Isolation of the barley powdery mildew resistance gene Mla opens the door to understanding the extensive allelic diversity of this locus. Completion of the Arabidopsis genome sequence enables the analysis of the complete set of R-gene homologues in a flowering plant. A new R gene, RPW8, conferring resistance in Arabidopsis to powdery mildew, reveals a new class of protein associated with pathogen recognition. New prospects for using R-gene polymorphism in agriculture are becoming apparent. PMID- 11418337 TI - Knowing the dancer from the dance: R-gene products and their interactions with other proteins from host and pathogen. AB - Cloning of plant disease resistance genes is now commonplace in model plants. Recent attention has turned to how the proteins that they encode function biochemically to recognize their cognate Avirulence protein and to initiate the disease-resistance response. In addition, attention has turned to how the Avirulence proteins of pathogens might alter susceptible hosts for the benefit of the pathogen, and what plant proteins might be required for that process. PMID- 11418338 TI - Nonhost resistance to Phytophthora: novel prospects for a classical problem. AB - Members of the oomycete genus Phytophthora are the most devastating pathogens of dicot plants. Recent developments in the study of these organisms have led to improved understanding of their phylogenetic relationships and trends in their evolution. Molecular analyses of nonhost (species-level) resistance offer exciting prospects for disease management. A model that evokes a complex interplay of several layers of specific resistance, mediated by a set of ancient broad-spectrum R-gene loci, is sufficient to explain existing cellular and molecular data on nonhost resistance to Phytophthora. PMID- 11418339 TI - Genes controlling expression of defense responses in Arabidopsis--2001 status. AB - In the past two years, the focus of studies of the genes controlling expression of defense responses in Arabidopsis has shifted from the identification of mutants to gene isolation and the ordering of genes within branches of the signal transduction networks. It is now clear that gene-for-gene resistance can be mediated through at least three genetically distinguishable pathways. Additional genes affecting salicylic-acid-dependent signaling have been identified, and double-mutant analyses have begun to reveal the order in which they act. Genes required for jasmonic-acid-dependent signaling and for induced systemic resistance have also been identified. PMID- 11418340 TI - Genetic dissection of systemic acquired resistance. AB - Significant progress has been made in the past year in understanding the mechanism of systemic acquired resistance. Mitogen-activated protein kinase cascades have been implicated as negative regulators of salicyclic acid accumulation and the induction of resistance. The salicylic acid signal is transduced through NPR1, a nuclear-localized protein that interacts with transcription factors that are involved in regulating salicylic-acid-mediated gene expression. Both promoter analyses and genetic studies have shown that gene expression in systemic acquired resistance requires not only the activation of a transcriptional activator(s) but also inhibition of a transcriptional repressor(s). Microarray experiments have been performed to search for those genes whose expression is transcriptionally regulated during systemic acquired resistance and to identify common promoter elements that control these genes. PMID- 11418341 TI - Fungal genomics and pathogenicity. AB - The filamentous fungal genetics community has enthusiastically embraced the utilization of genomics technologies to resolve long-standing issues in fungal biology. For example, such technologies have been proposed to study the mechanics of tip growth, photoreception, gene silencing, the molecular basis of conidiation, the pathway leading to sexual reproduction, and mechanisms of pathogenesis. These studies have provided a refreshing change of pace in research on filamentous fungi, which has lagged behind that on other eukaryotes in the exploitation of genome-wide methodologies. Despite the late start, several fungal genome sequencing projects are underway. The resulting databases will allow the comprehensive analysis of developmental processes that are characteristic of fungi, including the molecular nature of pathogenicity. DNA databases underpin analyses of the fungal transcriptome, proteome, and metabolome. This combined information will contribute to our basic understanding of not only the mechanics of infection but also the evolution of pathogenicity. PMID- 11418342 TI - Signal and nutrient exchange at biotrophic plant-fungus interfaces. AB - Biotrophic interfaces are formed in mutualistic and parasitic plant-fungus interactions. They result from coordinated developmental programs in both partners and represent specialized platforms for the exchange of information and nutritional metabolites. New data on the establishment and the components of functional interfaces have been obtained in a number of ways. First, by isolation of symbiotically defective mutants; second, by characterization of new genes and their products; and, third, by the identification and localization of components of biotrophic interfaces, such as cell-wall proteins, H+-ATPases and nutrient transporters. PMID- 11418343 TI - Genetics and genomics of root symbiosis. AB - Model genetics and genomics have been developed as tools for studying the third largest family of flowering plants, the Leguminosae, which includes important crop plants. Functional genomics strategies for the global analysis of gene expression, the elucidation of pathways and reverse genetics are established. These approaches provide new possibilities for investigating rhizobial as well as mycorrhizal endosymbiosis. Plant genes with central functions in these mutualistic interactions have been identified by positional cloning and gene tagging. With progress in Lotus japonicus genome sequencing, which was recently initiated by Japanese researchers, comparative genomics will contribute to our understanding of symbiosis, pathogenesis and the evolution of plant genomes. PMID- 11418344 TI - Rhizobium type III secretion systems: legume charmers or alarmers? AB - Mutagenesis and sequence analyses of rhizobial genomes have revealed the presence of genes encoding type III secretion systems. Considered as a machine used by plant and animal pathogens to deliver virulence factors into their hosts, this secretion apparatus has recently been proven to play a role in symbiotic bacteria leguminous plant interactions. PMID- 11418345 TI - Molecular basis of plant growth promotion and biocontrol by rhizobacteria. AB - Plant-growth-promoting rhizobacteria (PGPRs) are used as inoculants for biofertilization, phytostimulation and biocontrol. The interactions of PGPRs with their biotic environment, for example with plants and microorganisms, are often complex. Substantial advances in elucidating the genetic basis of the beneficial effects of PGPRs on plants have been made, some from whole-genome sequencing projects. This progress will lead to a more efficient use of these strains and possibly to their improvement by genetic modification. PMID- 11418346 TI - Merging molecular and ecological approaches in plant-insect interactions. AB - The singer-song-writer Paul Simon sang about the '50 ways to leave your lover'; plants have at least as many ways of coping with their insect herbivores. Recent research has elucidated the mechanisms of direct and indirect plant defenses, and has provided the first proof of a protective function for indirect defenses in nature. Insect attack elicits a large transcriptional reorganization that differs from that elicited by mechanical wounding. Elicitors in herbivore oral secretions can account for herbivore-specific responses. Patterns of transcriptional changes point to the existence of central herbivore-activated regulators of metabolism. PMID- 11418347 TI - Host-plant recognition by parasitic Scrophulariaceae. AB - Parasitic plants in the Scrophulariaceae invade the roots of neighboring plants in order to rob them of water and nutrients. A distinctive feature of these parasites is their ability to cue their development to small molecules released by host-plant roots. Evidence is continuing to emerge that parasite perception of host factors occurs via a redox-associated mechanism. Genes predicted to function during the early stages of parasite-host interactions have been cloned from both plant partners, and their characterization is providing a genetic framework on which to model subterranean plant-plant interactions. PMID- 11418348 TI - Diagnostic virology--the need for electron microscopy: a discussion paper. PMID- 11418349 TI - Clinically relevant sequence-based genotyping of HBV, HCV, CMV, and HIV. AB - The term 'genotyping' describes the genetic characterization of a genome. The genotype analysis is performed to identify mutations that differentiate one individual or strain from another. The mutations may confer resistance to specific antiviral drugs or they may simply allow classification of a strain as to 'type' and 'subtype'. There are four human viruses for which genotype information is clinically useful. Hepatitis B virus (HBV) infections are being treated with antiretroviral drugs and resistance after prolonged treatment is common. Since HBV cannot be cultured, the only method of detecting resistance conferring mutations in the genome is a genotypic analysis. Hepatitis C virus (HCV) infection can be cured by treatment with the combination of interferon and ribavirin but certain strains of virus are more resistant to treatment than others. The current recommendations are that all HCV type 1 infections be treated for 12 months whereas other types may be successfully treated in 6 months. Since interferon treatment may have significant side effects, the determination of HCV genotype is an important aspect of this therapeutic regimen. Treatment of cytomegalovirus (CMV) disease with nucleoside analogues occasionally results in resistant virus with mutations in the phosphotransferase gene (UL97) and/or the DNA polymerase gene (UL54) that can be tested with phenotypic or genotypic assays. Since CMV grows very slowly, it may be more clinically useful to perform a rapid genotypic assay although only the UL97 gene can be efficiently genotyped. Finally, the virus for which genotyping has become the standard of care, human immunodeficiency virus type 1 (HIV-1) can now be genotyped routinely by many clinical virology labs experienced with molecular amplification methods and automated DNA sequencing technology. All currently-available antiretroviral drugs are directed against either the protease or reverse transcriptase genes of HIV-1 and the mutations within these genes that confer resistance have been well described. Sequence-based genotyping methods are not necessarily the best approach for routine genotyping of these four viruses, but sequencing is the gold standard from which other methods are developed and against which they are compared. PMID- 11418350 TI - Evaluation of virus excretion by cells persistently infected with the bovine leukaemia virus (BLV) using monoclonal antibodies. AB - BACKGROUND: bovine leukaemia virus (BLV) is the causative agent of enzootic bovine leukaemia. Studies in vitro usually require the use of infected cell lines, mostly to produce antigen. Two of the most widely used cell lines are FLK BLV and BLV-bat2. OBJECTIVE: the dynamics of the excretion of BLV proteins and whole virus by the persistently BLV-infected cell lines mentioned above was studied using an indirect ELISA in combination with eight monoclonal antibodies (mAbs) and cow and rabbit serum. STUDY DESIGN: tissue culture flasks were seeded with different concentrations of cells (13000-67000 cells per cm2, corresponding to 1-5 million cells per 75 cm2 flask) and were studied for 20 days. Samples (1.5 ml) were removed every 24 h and the presence of BLV proteins was determined using an indirect ELISA assay in which the antigen reaction with the monoclonal antibodies was evidenced by peroxidase labeled anti-mouse immunoglobulins. RESULTS: cell line FLK-BLV produced a complete monolayer as early as 4 days after passage, 3 days earlier than BLV-bat2. Using mAbs, the amount of viral proteins in the supernatant showed a cyclic pattern, with two evident peaks at days ca. 8 and 16. These peaks occurred even in the absence of passage or medium change, which causes depletion of essential nutrients and acidity. In comparison to polyclonal serum, mAbs gave more clear and defined values and are useful for determining the dynamics of viral production. CONCLUSION: when aiming for high viral yield, BLV should be harvested between days 6 and 8 after passage, when viral shedding is at its maximum. These results are very useful for preparing antigen for monoclonal antibody production, or for techniques such as ELISA or Western blot. PMID- 11418351 TI - A peptide enzyme linked immunosorbent assay (ELISA) for the detection of human immunodeficiency virus type-2 (HIV-2) antibodies: an evaluation on polymerase chain reaction (PCR) confirmed samples. AB - BACKGROUND: HIV-1 and HIV-2 infections differ in prognosis, and may also require different prevention and/or treatment approaches. Thus, estimating the true prevalence of HIV-1 and HIV-2 infections, as well as co-infections, is a critical step in controlling the disease. There are a few commercial ELISA and immunoblot kits, which can differentiate between HIV-1 and HIV-2 infections. However, some of these assays overestimate the prevalence of dual infection. Hence, it is necessary to develop assays capable of discriminating between the two infections. OBJECTIVES: To develop a synthetic HIV-2 env based peptide ELISA for the detection of HIV-2 specific antibodies and evaluate its performance on samples from HIV positive individuals previously tested by HIV-1 and HIV-2 PCR and HIV seronegative individuals. STUDY DESIGN: We studied 45 HIV seronegative and 63 HIV infected individuals, including 30 HIV-1 PCR and immunoblot positives, 19 HIV-2 PCR and immunoblot positives, five HIV-1 and two PCR and dual immunoblot positives, two PCR negative but positive for HIV-2 by immunoblot and seven dual immunoblot positives who were only positive for HIV-1 by PCR. RESULTS: All 24 HIV 2 PCR positive samples tested were positive by the peptide assay. Among 30 HIV-1 PCR and immunoblot positive samples, only one (3.3%) showed an absorbance value above the cut off level. The seven dual positive samples by immunoblot (only positive for HIV-1 by PCR) were negative by the HIV-2 peptide ELISA. There was a 100% concordance between HIV-2 PCR and peptide ELISA. The sensitivity, specificity, and the likelihood ratio for the peptide ELISA were 100,94.9, and 19.5, respectively when compared against the PCR findings. CONCLUSIONS: This ELISA, using a specific immunodominant epitope (11 amino acids) from the transmembrane (gp36) portion of the HIV-2 envelope glycoprotein showed a high concordance with PCR findings. This can be considered as a highly sensitive, specific and economically feasible assay for the discrimination of HIV-1 and HIV 2, and may serve as an alternative to HIV-2 PCR in epidemiological studies. PMID- 11418352 TI - Maturation of IgG avidity to individual rubella virus structural proteins. AB - BACKGROUND: the structural proteins of rubella virus, the capsid protein C and the envelope glycoproteins E1 and E2 were produced in lepidopteran insect cells using baculovirus expression vectors. The C-terminal ends of the corresponding proteins were fused to a polyhistidine tag for easy and gentle purification by metal ion affinity chromatography. OBJECTIVES: to investigate the maturation of natural and vaccinal IgG avidity against individual authentic and recombinant rubella virus (RV) structural proteins. STUDY DESIGN: the analysis was carried out using a modified immunoblotting technique where the purified baculovirus expressed proteins were compared with authentic rubella virus proteins. Altogether, 47 well-characterised serum samples from both naturally infected patients and vaccines were studied. RESULTS: after natural RV infection, IgG antibodies specific for the E1 protein were predominant not only in terms of levels, but also in terms of rate and magnitude of avidity maturation. The avidity development of the IgG antibodies was much slower in vaccines than in patients after a natural RV infection. CONCLUSIONS: together, our results indicate that IgG avidity determination in conjunction with immunoblot analysis is useful in the diagnosis of a RV infection. The recombinant proteins showed similar reactivity patterns in the immunoblot analyses as compared with the authentic viral structural proteins, suggesting suitability for serodiagnostics. PMID- 11418353 TI - Baculovirus expression of parvovirus B19 (B19V) NS1: utility in confirming recent infection. AB - BACKGROUND: The presence of anti-parvovirus B19 (B19V) IgM against viral capsid proteins (VP1 and VP2) has long been used to detect recent infection. The utility of antibodies directed against B19V NS1 protein has received less attention as a serological indicator of recent infection, although anti-B19V NS1 IgG has been associated with persistent infection. OBJECTIVES: To elucidate the role of anti B19V NS1 antibody detection in recent infection, full-length B19V NS1 was expressed and purified. The resultant antigen was used to develop both Western blot assays and microplate ELISA for the detection of NS1 antibodies. STUDY DESIGN: Serum specimens were obtained from individuals recently infected with B19V (children (n=16), adults (n=40)) and from 17 individuals with no evidence of recent B19V infection. All specimens were screened for anti-B19V NS1 IgG and IgM. RESULTS: It was observed that 68.8% (11/16) of children recently infected with B19V were anti-B19V NS1 IgG seropositive. Furthermore, 27.5% (11/40) anti-B19V VP2 IgM positive specimens also contained anti-B19V NS1 IgM when tested by ELISA, while no reactivity was observed following Western blot analysis, possibly due to the absence of conformational epitopes. CONCLUSIONS: Anti-B19V NS1 IgM detection may have utility in the confirmation of recent infection with B19V. PMID- 11418354 TI - Monitoring response during a randomised controlled trial of escalating interferon dose for chronic hepatitis C infection: predictive value of quantitative and qualitative HCV RNA assays. AB - BACKGROUND: In chronic hepatitis C infection, raising the interferon dose in initial non-responders may increase the generally poor sustained response rates. Monitoring virological response is essential in this kind of individual patient based approach. Quantitative HCV RNA assays are increasingly used for this purpose. However, their additional value as compared to strictly qualitative HCV RNA assays should be evaluated before they are implemented as a routine measurement, since these assays are more expensive and time consuming than qualitative assays. OBJECTIVES: Goals of this study were (1) to test the hypothesis that increasing interferon dose in initial non-responders results in permanent viral clearance in more patients and (2) evaluation of the predictive value of quantitative versus qualitative HCV RNA assays before and during treatment. STUDY DESIGN: 63 patients were treated in a randomised controlled trial of escalating interferon dose. In the standard treatment group patients received 6 MU alpha-2a thrice weekly for 3 months followed by 3 MU thrice weekly for 3 months. In the experimental group interferon dose was escalated at 6 weeks to 9 MU if HCV RNA was still detectable at 4 weeks. Predictors of response were analyzed at various time points before and during treatment and the predictive value of quantitative HCV RNA measurements was compared to that of qualitative HCV RNA assays. RESULTS: No significant difference in sustained response rate was found between the treatment groups at the end of follow-up. At baseline, the strongest independent predictor for a sustained response was a viral load level below 10(6) copies/ml and age younger than 40 years. During treatment a negative HCV RNA status at week 4 was the strongest predictor of a sustained response. Viral load levels during treatment did not independently predict a sustained response. CONCLUSIONS: While on treatment, qualitative HCV RNA assays should be used to monitor response. PMID- 11418355 TI - Antiviral drugs: current state of the art. AB - The chemotherapy of virus infections has definitely come of age. There are now 15 antiviral agents that have been formally licensed for the treatment of human immunodeficiency virus infections (zidovudine, didanosine, zalcitabine, stavudine, lamivudine, abacavir, nevirapine, delavirdine, efavirenz, saquinavir, ritonavir, indinavir, nelfinavir, amprenavir, lopinavir) and several others, such as tenofovir disoproxil, emtricitabine, capravirine, emivirine, T-20 (pentafuside) and AMD3100 (bicyclam) are under clinical development. Lamivudine has been approved, and several other compounds (such as adefovir dipivoxil, emtricitabine and entecavir) are under clinical development, for the treatment of hepatitis B virus infections. Among the anti-herpesvirus agents, aciclovir, valaciclovir, penciclovir, famciclovir, idoxuridine, trifluridine and brivudin are used in the treatment of herpes simplex virus and varicella-zoster virus infections, and ganciclovir, foscarnet, cidofovir, fomivirsen and maribavir (the latter in the developmental stage) are used in the treatment of cytomegalovirus infections. Following amantadine and rimantadine, the neuraminidase inhibitors, zanamivir and oseltamivir, have now become available for the therapy and prophylaxis of influenza virus infections, and so is ribavirin for the treatment of respiratory syncytial virus infections and the combination of ribavirin with interferon-alpha for the treatment of hepatitis C virus infections. PMID- 11418356 TI - Comparison of systems performance for TT virus detection using PCR primer sets located in non-coding and coding regions of the viral genome. AB - BACKGROUND: the heterogeneity of the TT virus (TTV) DNA prevalence values reported from comparable human cohorts suggests that diagnostic PCR protocols still require to be optimized. OBJECTIVES: to design TTV PCR primer sets with low genotype restriction and to compare their performances with commonly used amplification systems. STUDY DESIGN: we compared full length TTV genomic sequences and identified conserved nucleotide patterns in the 5' and 3' non coding regions of the viral genome. This permitted to design two new primer sets usable for the PCR amplification of the most divergent human isolates of TTV described to date. The performances of these amplification systems were compared with those of three other PCR systems earlier used for prevalence studies. RESULTS: the primer systems P5Bx and P3Bx exhibited higher PCR scores than the other systems tested; 14 to 34% improvement values were obtained, and divergent positive results of earlier described PCR systems were confirmed systematically by our new detection assays. CONCLUSIONS: an optimized detection of TT virus DNA is a pre-requisite for the accurate epidemiological survey of viral infection and for the realization of phylogenetic studies. Such PCR systems with low genotype restriction will be helpful in the future for a better knowledge of natural history of TT virus infection. PMID- 11418357 TI - R-Mix cells are faster, at least as sensitive and marginally more costly than conventional cell lines for the detection of respiratory viruses. AB - OBJECTIVE: To evaluate shell vials of R-Mix, a combination of mink lung cells and human adenocarcinoma cells (strains Mv1Lu and A549, respectively, Diagnostic Hybrids, Athens, OH) to detect respiratory viruses from prospective clinical respiratory specimens and frozen stocks. STUDY DESIGN: We compared the performance of R-Mix to conventional culture (CC) using tubes of PMK, Hep-2, and MRC5 to detect respiratory viruses from fresh clinical specimens. For each respiratory specimen submitted to virology, two shell vials of R-Mix were inoculated and examined twice (generally after 24 and 48 h) by an indirect test with a pool of immunofluorescent antisera to influenza A and B, adenovirus, parainfluenza 1-3 and RSV (DAKO, Carpinteria, CA). If positive, testing with monoclonal antisera was done. CCs were incubated for 10 days, examined daily for cytopathological effect, hemadsorbed twice and stained if positive. Cost comparison was done. Lastly, respiratory viruses frozen from previous years were inoculated onto R-Mix. RESULTS: R-Mix was positive for all 29 frozen virus stocks. In the clinical trial, 396 prospective specimens were inoculated into R Mix and CC. R-Mix identified 21 specimens as respiratory virus positive; CC identified 19. Turn-around time of R-Mix for positive specimens was 1.4 days; for CC it was 5.2 days. Turn-around time of R-Mix for all specimens (positive and negative) was 2.0 days; for CC it was 9.8 days. The overall cost of R-Mix was approximately 11% more than that of CC. CONCLUSION: R-Mix enabled rapid identification of all the frozen virus stocks representing the seven major respiratory viral groups. When compared to CC, R-Mix was slightly more sensitive than three cell lines (four tubes) used in CC but it was several days faster. PMID- 11418358 TI - Comparison of plasma polymerase chain reaction and pp65-antigenemia assay in the quantification of cytomegalovirus in liver and kidney transplant patients. AB - BACKGROUND: Cytomegalovirus (CMV) is a significant problem in transplantation. The antiviral treatment is based on the clinical symptoms and the rapid laboratory diagnosis. Although polymerase chain reaction (PCR) methods have already been widely used, the clinical correlation of the findings is not clear. OBJECTIVE: The objective of this study was to investigate the usefulness of a quantitative plasma PCR test and compare it with the pp65-antigenemia test in the detection of clinically significant CMV infections in liver and kidney transplant patients. STUDY DESIGN: The clinical material consisted of 253 consecutive blood samples was tested using a quantitative polymerase chain reaction test, Cobas Amplicor CMV Monitor (Roche) and pp65 antigenemia assay. Plasma was used for PCR and leucocytes were used for the antigenemia test. RESULTS: CMV was detected in 89 out of 253 blood samples by one or both methods. PCR detected 78 (range 274 165000 copies/ml) and pp65 antigenemia test 79 (range 1-1500 positive cells/50000) of the positive findings. The sensitivity and specificity of PCR test was 86 and 94%, respectively. The PCR detected all clinically significant CMV infections (>10 positive cells in pp65 test) and infections which required antiviral treatment. In addition, the correlation between the two tests was almost linear. CONCLUSIONS: The quantitative PCR appears to be a suitable alternative to diagnose and monitor CMV infections in transplant patients. PMID- 11418359 TI - Human papillomavirus infection, risk for subsequent development of cervical neoplasia and associated population attributable fraction. AB - BACKGROUND: Human papillomavirus (HPV) is the major cause of cervical neoplasia but estimates of the population attributable fraction (PAR%), of HPV vary. PAR% has not been derived from longitudinal studies although assessment of HPV exposure prior to the neoplasia diagnosis should increase validity of such estimates. AIMS: Systematic review and meta-analysis of longitudinal studies on HPV associated relative risk (RR) for and PAR% of HPV16 in cervical neoplasia. METHODS: Pertinent data from longitudinal studies was made available through Medline and substituted by various hand searches. HPV associated weighted mean RR, with 95% confidence interval (CI) of cervical neoplasia, and the PAR% of HPV16 in cervical carcinoma were estimated both for unselected and low HPV prevalence populations. RESULTS: HPV associated RR of cervical carcinoma in PCR based studies was 17 (95% CI 8.2-33). HPV16 associated RRs in seroepidemiological studies were 3.3 (95% CI 2.2-4.9) for the unselected population, HPV16 seroprevalence 11.0%, and 12.5 (95% CI 5.5-29) for a population with low HPV16 seroprevalence of 5.3%. Corresponding PAR% estimates of HPV16 were 27 and 44%, respectively. CONCLUSION: Protective vaccination against HPV16 infection would prevent up to 44% of cervical carcinoma. PMID- 11418360 TI - Dynamics of viral quasispecies during interferon therapy in non responder chronic hepatitis C patients. AB - BACKGROUND: the reference method to study the HCV complexity was cloning and sequence analysis of a sufficient number of clones. The evolution of the viral complexity in chronic non responder patients during treatment with standard doses of interferon was not very well investigate because this method was expensive and labour intensive when large series of patients were concerned. Meanwhile, with the alternative Single-Strand Conformation Polymorphism (SSCP) method, a rough estimation of the quasispecies present in a given sample could be obtained. OBJECTIVES: the aim of the study was to analyse the evolution of HCV heterogeneity, investigated by SSCP analysis targeted to the HVR-1, in 30 nonresponders chronic hepatitis C patients treated by Interferon-alpha 3MUI. RESULTS: genotype 1 was the main HCV type found in this population (77% of non responder patients). Before treatment, the SSCP assay revealed a high complexity pattern: the median of SSCP band number was 9. During IFN-alpha treatment, SSCP band number didn't change. However a significant decrease of the viral load was observed (P<0.01). Patients with variations in their SSCP patterns after therapy significantly decreased HCV RNA levels (P<0.002). In one third of patients the SSCP profile didn't change at all. CONCLUSIONS: we observed that viral heterogeneity didn't change in non responder chronic hepatitis C patients during IFN-alpha treatment. Nevertheless patients with a low number of pre-treatment quasispecies exhibited an improvement of the response (P<0.02). These phenomena were probably due to a selection of resistant variants present prior onset of therapy. PMID- 11418361 TI - Outbreak of Hepatitis B among injecting drug users in Denmark. AB - BACKGROUND: The incidence of hepatitis B is low in Denmark, but injecting drug users (IDUs) remains a high-risk group for this infection. OBJECTIVES: The aim of the study was to describe a hepatitis B outbreak among IDUs by comparing existing registers. Additionally, we wanted to analyze the genetic variation of the hepatitis B virus involved in the outbreak. STUDY DESIGN: In the County of Funen, registers of laboratory diagnosis, hospital records and reports from clinicians to the Medical Officer of Health (MOH) were compared between 1992 and 1998. HBsAg positive sera recovered from the epidemic were sequenced and compared to known HBV strains. RESULTS: We identified 648 cases of hepatitis B of which 51% (332) were acute infections. The laboratory database identified 96% (319/332) of these, 45% (150/332) were admitted to hospital and 38% (127/332) were reported to public health. By capture-recapture analysis based on MOH reports and hospital records the estimated total number of acute cases were 334 (95% C.I. 283-385). We sequenced 75 HBsAg positive samples and identified two very similar strains of genotype D (serotype ayw3) among IDUs involved in the outbreak. CONCLUSIONS: The current surveillance system did not detect the majority of acute hepatitis B cases in County of Funen. We suggest laboratory-based surveillance of hepatitis B to be implemented at a national level as this may identify new outbreaks faster and more complete than the current surveillance system. PMID- 11418362 TI - Occurrence of HIV-1 reverse transcriptase gene mutation at codon 215 in HIV infected infants. AB - BACKGROUND: The nucleoside reverse transcriptase inhibitor Zidovudine (ZDV) decreases mother to child transmission of HIV infection. Nevertheless, significant proportions of mothers who are treated during pregnancy with ZDV still transmit the virus. Along with other factors, failure of ZDV prophylaxis may be due to maternal infection with ZDV-resistant strains and their consequent vertical transmission. OBJECTIVES: The purpose of the study was to investigate the occurrence of mutations at codon 215 in HIV-1 infected infants and its association with clinical status and virological parameters. STUDY DESIGN: The cohort consisted of 49 HIV-1 infected infants. Mononuclear cell DNA was isolated from whole blood and served as the input DNA for both a qualitative DNA PCR and the codon 215 assay (nested PCR). HIV-1 viral load (RNA PCR) was measured in plasma by the Roche Amplicor Monitor assay. RESULTS: Twelve of the 49 (24.5%) demonstrated viral strains with mutation at codon 215. A significant difference was found in infants born between 1992 and 1994 (6.3%) compared to those born in 1998-1999 (33.3%). Furthermore, in those infants born in 1998-1999, there was a trend toward an increase in the frequency of zidovudine resistant mutations at codon 215 corresponding to an increase in maternal zidovudine treatment. The mixture of wild and mutant HIV-1 strains was detected in two of 14 infants (14.3%) with a low viral load (<750000 c/ml) compared to nine of 19 (47.4%) infants with extremely high levels of HIV-1 RNA concentration (>750000 c/ml). Only two of 33 tested infants were HIV symptomatic and in both, a mixture of wild and mutant HIV-1 strains was detected. In both infants, the viral load was >750000 c/ml. CONCLUSIONS: This study showed that the overall frequency of ZDV resistant strains in infants born in 1998-1999 was significantly higher than that found in infant samples from 1992 to 1994. By 1998, the standard of care for mothers known to be infected with HIV was treatment with either monotherapy using ZDV or combined therapy using a variety of antiretroviral agents including ZDV. Although the exact role of ZDV resistance in limiting the effectiveness of therapies aimed at blocking vertical transmission of HIV remains to be defined, it is clear that drug resistant strains of HIV are occurring more frequently in all types of HIV infection, including infants. PMID- 11418363 TI - Spanish contributions to management of acute promyelocytic leukemia. PMID- 11418364 TI - The cyclin D1 alternative transcripts [a] and [b] are expressed in normal and malignant lymphocytes and their relative levels are influenced by the polymorphism at codon 241. AB - BACKGROUND AND OBJECTIVES: The cyclin D1 gene, CCND1, is alternatively spliced to produce transcripts [a] and [b] in a manner apparently modulated by a polymorphism (A/G) at codon 241. Studies have indicated that the polymorphism can affect the prognosis of patients with different types of solid tumors. This study aimed to determine the relative levels of transcripts [a] and [b] in normal and malignant peripheral blood mononuclear cells (PBMNC), and to investigate whether these were influenced by the polymorphism. The impact of the polymorphism on the survival of a group of mantle cell lymphoma (MCL) patients was also to be studied. DESIGN AND METHODS: The polymorphism was genotyped, using restriction fragment length polymorphism analysis, in 74 patients (42 MCL, 19 chronic lymphocytic leukemia, 13 normal controls) and the relative level of transcripts [a] and [b] determined using a competitive reverse transcription polymerase chain reaction method. Kaplan-Meier survival curves and the log-rank test were used to analyze the survival data. RESULTS: Of the cases genotyped, 39 were heterozygous for the polymorphism, 24 homozygous G and 11 homozygous A. Both transcripts [a] and [b] were expressed in normal PBMNC and malignant lymphocytes, with the polymorphism affecting their relative levels. Neither the predominant transcript, nor genotype, significantly influenced survival of the MCL patients studied. INTERPRETATION AND CONCLUSIONS: Contrary to previous reports, patients who were homozygous A at the polymorphism produced more transcript [a] whilst homozygous G patients had more transcript [b]. In the small cohort studied, the polymorphism did not appear to affect the prognosis of the patients with MCL. PMID- 11418365 TI - Variability in the levels of PML-RAR alpha fusion transcripts detected by the laboratories participating in an external quality control program using several reverse transcription polymerase chain reaction protocols. AB - BACKGROUND AND OBJECTIVES: The detection of PML-RAR by reverse transcription (RT) polymerase chain reaction (PCR) in acute promyelocytic leukemia (APL) patients who are in hematologic remission influences therapeutic decision making in several trials. In the light of this, the Spanish group has recently designed an external quality assessment program (EQAP) of RT-PCR detection of PML-RAR, which includes a study of sensitivity of the participating laboratories. DESIGN AND METHODS: Eighteen laboratories were involved in the program. Ten laboratories followed the method of Biondi et al., 5 employed that of Borrow et al. and the 3 remaining used other protocols. The sensitivity was studied in five rounds of quality control. The first two shipments consisted of dilutions of NB4 RNA into non-APL RNA. The third round consisted of serial dilutions of the NB4 cell line into HL60 cells. The fourth and five rounds consisted of plasmid dilutions containing the bcr1 and bcr3 PML-RAR isoforms. RESULTS: The results showed that the distinct methods allow detection of the PML-RAR hybrid up to a dilution of 10(-4), and exceptionally, up to 10(-5). The laboratories following the method of Biondi et al. usually detected the 10(-3) dilution and less frequently the 10(-4) one, whereas those using other methods usually detected PML-RAR transcript in the 10(-4) dilution, and less commonly in the 10(-5) dilution. However, each of the PCR methods used by EQAP participating laboratories successfully detected at least 50 copies of PML-RAR alpha fusion transcript in plasmid dilution controls. INTERPRETATION AND CONCLUSIONS: The results point to heterogeneous sensitivity amongst participating laboratories. This may reflect differences in methodology, although variations in sample quality may also account for discrepant findings. PMID- 11418366 TI - The PML-RAR alpha transcript in long-term follow-up of acute promyelocytic leukemia patients. AB - BACKGROUND AND OBJECTIVES: Detection of PML-RAR alpha transcripts by RT-PCR is now established as a rapid and sensitive method for diagnosis of acute promyelocytic leukemia (APL). Although the majority of patients in long-term clinical remission are negative by consecutive reverse transcription polymerase chain reaction (RT-PCR) assays, negative tests are still observed in patients who ultimately relapse. Conversion from negative to positive PCR has been observed after consolidation and found to be a much stronger predictor of relapse. This study reports on 47 APL patients to determine the correlation between minimal residual disease (MRD) status and clinical outcome in our cohort of patients. DESIGN AND METHODS: The presence of PML-RAR alpha t transcripts was investigated in 47 APL patients (37 adults and 10 children) using a semi-nested reverse transcriptase-polymerase chain reaction to evaluate the prognostic value of RT PCR tests. RESULTS: All patients achieved complete clinical remission (CCR) following induction treatment with all-trans retinoic acid (ATRA) and chemotherapy (CHT) or ATRA alone. Patients were followed up between 2 and 117.6 months (median: 37 months). Relapses occurred in 11 patients (9 adults and 2 children) between 11.4 and 19 months after diagnosis (median: 15.1 months) while 36 patients (28 adults and 8 children) remained in CCR. Seventy-five percent of patients carried the PML-RAR alpha long isoform (bcr 1/2) which also predominated among the relapsed cases (9 of 11) but did not associate with any adverse outcome (p= 0.37). For the purpose of this analysis, minimal residual disease tests were clustered into four time-intervals: 0-2 months, 3-5 months, 6-9 months and 10-24 months. INTERPRETATION AND CONCLUSIONS: Children showed persisting disease for longer than adults during the first 2 months of treatment. At 2 months, 10 (50%) of 20 patients who remained in CCR and 4 (80%) of 5 patients who subsequently relapsed were positive. Patients who remained in CCR had repeatedly negative results beyond 5.5 months from diagnosis. A positive MRD test preceded relapse in 3 of 4 tested patients. The ability of a negative test to predict CCR (predictive negative value, PNV) was greater after 6 months (>83%), while the ability of a positive test to predict relapse (predictive positive value, PPV) was most valuable only beyond 10 months (100%). This study (i) highlights the prognostic value of RT-PCR monitoring after treatment of APL patients but only from the end of treatment, (ii) shows an association between conversion to a positive test and relapse and (iii) suggests that PCR assessments should be carried out at 3-month intervals to provide a more accurate prediction of hematologic relapses but only after the end of treatment. PMID- 11418367 TI - Early and delayed consolidation chemotherapy significantly improves the outcome of children with intermediate risk acute lymphoblastic leukemia. Final results of the prospective randomized PETHEMA ALL-89 TRIAL. AB - BACKGROUND AND OBJECTIVES: To evaluate the impact of early and delayed consolidation chemotherapy on the outcome of children with acute lymphoblastic leukemia (ALL) stratified according to risk groups. DESIGN AND METHODS: From 1989 to 1994, 195 children (< or = 15 years old) diagnosed as having ALL (ALL-L3 excluded) in 15 Spanish hospitals entered the prospective, randomized PETHEMA ALL 89 trial. Patients were stratified into low-risk (LR), intermediate-risk (IR) and high-risk (HR) groups according to their initial features and the rate of response to induction therapy. LR-ALL patients were randomized to receive or not early consolidation chemotherapy (C-1). After receiving C-1, IR patients were randomized to receive or not delayed consolidation chemotherapy (C-2). HR patients received C-1 and C-2 chemotherapy. Standard maintenance chemotherapy was administered to all patients for 2 years. High doses of intravenous methotrexate and 12 triple intrathecal doses were given as prophylaxis against central nervous system (CNS) disease. RESULTS: The mean (and standard deviation) age was 6 (4) years and 120 patients were males. Fourteen patients had early pre-B-ALL, 149 common or pre-B-ALL, and 32 T-ALL. Complete remission (CR) was attained in 189 patients (97%), 11 of whom (6%) had a slow response. Risk group stratification after CR was: LR 89, IR 50 and HR 56 patients (including a subset of 26 patients at very high risk). Ten-year event-free survival (EFS) and overall survival (OS) probabilities for the whole series were 58% (95% CI: 52-64%) and 69% (61-77), respectively, with a median follow-up of 8.7 years. Dividing the patients according to risk group, the 10-year EFS and OS probabilities in the LR group were 71% (63-79) and 86% (80-92), respectively; in the IR group 69% (57-81) and 76% (64-88), respectively, and in the HR group 30% (18-42) and 44% (32-57), respectively. For LR patients receiving C-1, EFS and OS were 79% (57-92) and 90% (82-98), respectively, versus 62% (48-76) and 66% (51-81) in patients not receiving C-1 (p= 0.06). For IR patients, EFS and OS were significantly improved in those receiving early and delayed consolidation (EFS 87% (74-88) vs. 52% (41 70), and OS 92% (87-97) vs. 61% (51-71)(p=0.036). Prognostic factors for EFS identified in multivariable analyses were: age >10 years in the LR group (OR 3.5, 95% CI 1.3-9.5, p=0.01), and treatment with C-2 in IR patients (OR 5.0, 95% CI 1.4-17.8, p=0.01). The CNS relapse rate was 4% for all the series (including the HR subset). Tolerance to treatment was good. INTERPRETATION AND CONCLUSIONS: In this study, early consolidation seemed to improve the prognosis of children with LR-ALL, but differences in EFS were not significant. Delayed consolidation had a favorable influence on the outcome of IR-ALL. CNS preventive treatment without cranial irradiation was effective in all the groups of ALL patients. PMID- 11418368 TI - Ten-year follow-up of a single center prospective trial of unmanipulated peripheral blood stem cell autograft and interferon-alpha in early phase chronic myeloyd leukemia. AB - BACKGROUND AND OBJECTIVES: The potential role of autologous stem cell transplantation (ASCT) as an alternative therapeutic strategy in chronic myelogenous leukemia (CML) has been widely explored in pilot studies, but the clinical results in terms of survival have so far been evaluated only retrospectively and in heterogeneous groups of patients. The goal of our prospective study was to evaluate the feasibility and long-term efficacy of unmanipulated ASCT followed by low-dose interferon-alpha in a homogeneous group of patients affected by CML in a very early phase of disease. DESIGN AND METHODS: Twenty-six unselected consecutive patients with CML in chronic phase underwent stem cell collection at diagnosis, then received cytoreductive treatment with hydroxyurea and, subsequently, a busulphan-melphalan myeloablative regimen followed by unmanipulated stem cell graft within one year of diagnosis. Interferon was given a median of 6.5 months after transplant at escalating doses, starting from 0.5 x 10(6) IU 3 times/week, on the basis of the clinical and hematologic tolerance. RESULTS: Median chronic phase duration from diagnosis is 9 years. The ten-year projected probability of overall survival from diagnosis is 55% with a median follow-up of surviving patients of 9.5 years (8-10.5); median survival has not been reached after ten years. INTERPRETATION AND CONCLUSIONS: Our experience suggests that high-dose therapy followed by unmanipulated peripheral blood stem cell transplantation and low-dose interferon-alpha is a feasible approach, which results in long-term survival in newly diagnosed CML patients. These data need to be confirmed in controlled trials comparing ASCT with other therapeutic approaches, such as the use of interferon-alpha alone or in combination with other agents. PMID- 11418369 TI - Treatment of low grade gastric mucosa-associated lymphoid tissue lymphoma in stage I with Helicobacter pylori eradication. Long-term results after sequential histologic and molecular follow-up. AB - BACKGROUND AND OBJECTIVES: Most cases of gastric low-grade mucosa-associated lymphoid tissue (MALT) lymphoma are associated with H. pylori. In localized disease (stage I), eradication of H. pylori can result in histologic regression of the lymphoma in 50% to 100% of the patients. Moreover, in half of the apparently cured patients a monoclonal rearrangement of the IgH gene can be demonstrated. However, data on the long-term outcome of the patients are scarce. We report the evolution of a series of patients followed-up since 1994 in order to evaluate the long-term outcome of the apparently cured lymphoma. DESIGN AND METHODS: From January 1994 to July 2000, 19 consecutive patients with stage I gastric low grade MALT lymphoma were sequentially studied in our hospital. They had all been diagnosed by endoscopy and had had a complete staging (including CT scan, contrast X-ray of the small bowel, bone marrow biopsies, immunophenotyping of bone marrow and peripheral blood and, in the later years, endoscopic ultrasonography). Diagnosis required established histologic criteria for low grade MALT lymphoma in the samples obtained by endoscopy. The investigation of H. pylori status included histologic search, serology and breath test urea-(13)C. Only patients in stage I disease associated with H. pylori were included in the study. Patients received standard triple therapy for eradication of H.pylori and after treatment were sequentially followed-up with endoscopies performed every 2 3 months in the first year, every 6 months in the second year and then yearly. Post-treatment biopsies were obtained by endoscopy for histologic studies, H. pylori cultures and molecular studies. The criteria of Wotherspoon et al. were used for the histological evaluation. Molecular studies were performed with a polymerase chain reaction analysis of the IgH gene using semi-nested procedures with consensus primers for the V(H) (Fr3A/Fr2A) and J(H) (LJH and VLJH) regions. RESULTS: After the eradication treatment, 18 of the 19 patients (94.7%) achieved histologic regression of the MALT lymphoma that occurred after a mean of 4.6 months (range 2-19). In 11 of the 18 histologically cured patients (61%) a monoclonal rearrangement of the IgH gene was demonstrated. In 2 patients the monoclonality disappeared completely, but 9 of the 11 patients (82%) had either persistent (3 patients) or intermittently persistent (5 patients) monoclonality for as long as 64 months. None of the patients who achieved a histologic remission (either with or without monoclonality) relapsed after a mean follow-up of 37 months (range 2-78). Two patients were lost to follow-up and another patient died of a gastric carcinoma; the remaining 15 patients are still in histologic remission after a mean period of 43 months (range 5-78). Ten patients studied between 1994 and the end of 1996 are in remission after a mean of 59 months (range 33-78). INTERPRETATIONS AND CONCLUSIONS: In most cases of gastric low-grade MALT lymphoma in stage I eradication of H. pylori can produce histologic regression of the lymphoma and this regression can be maintained for years. However, IgH gene monoclonality can be detected and persists in most cases. Although this persistent monoclonality seems to indicate the presence of a latent lymphoma population, over a period of 6 years it has not so far influenced the outcome. These findings indicate that in cases of localized gastric low-grade MALT lymphoma associated with H. pylori, the first step of treatment should be eradication of the H. pylori; however, a close and long follow-up is essential to determine the ultimate outcome of these patients and the possible significance of the persistent monoclonality. PMID- 11418370 TI - Anti-CD20 antibody (rituximab) administration in patients with late occurring lymphomas after solid organ transplant. AB - BACKGROUND AND OBJECTIVES: Aggressive diffuse large cell non-Hodgkin's lymphoma (DLCL) occurring late after a solid organ transplant fails to regress after discontinuation of immunosuppression. Moreover, chemotherapy treatment is associated with a high mortality rate due to severe toxicity. Since the majority of post-transplant lymphoproliferative disorders derive from B-lineage lymphocytes, the administration of anti-B monoclonal antibodies represents a rational therapeutic option. DESIGN AND METHODS: Five patients who developed CD20 positive DLCL more than two years after heart or liver transplantation were treated with a weekly chemotherapy program (2 patients), radiotherapy (2 patients) and surgery (1 patient) followed by a minimum of 4 intravenous doses of rituximab (375 mg/m(2)). RESULTS: A favorable clinical outcome was observed in three patients in whom surgery or radiotherapy had produced significant tumor debulking. Only a partial clinical effect was documented in the two patients with advanced clinical stage disease. INTERPRETATION AND CONCLUSIONS: Rituximab can be safely administered to patients with aggressive CD20-positive DLCL occurring late after a solid organ transplant. However, a positive clinical outcome may be expected only in patients in whom surgery or radiotherapy has achieved significant regression of tumor burden. PMID- 11418371 TI - Successful pregnancies following an egg donation program in women with previously treated Hodgkin's disease. AB - BACKGROUND AND OBJECTIVES: In order to draw attention not only to patients affected by a neoplasia, but also to those who may have problems of sterility, we describe six women affected by Hodgkin's disease who had precocious menopause due to chemotherapy and/or radiotherapy but who were safely delivered of children. These pregnancies were achieved through oocyte donation, in vitro fertilization and intrauterine embryo transfer or oocyte intracytoplasmic insemination. DESIGN AND METHODS: During natural or iatrogenic menopause, the uterus preserves its capacity to respond to steroidal hormones and to permit implantation and development of an embryo. Our study concerns six young females with iatrogenic menopause caused by treatment of Hodgkin's disease who carried a pregnancy to term. The pregnancies were achieved by oocyte donation, in vitro fertilization and intrauterine embryo transfer or oocyte intracytoplasmic insemination. Endometrial maturation was obtained by administration of estradiol and progesterone. Steroidal therapy was administered until the 13th-14th week in relation to placental function. RESULTS: Five of the 6 females underwent Caesarean section because of a twin birth or complications during the third trimester of pregnancy (gestosis). All the delivered children are, to date, well; their median age is 4 years. INTERPRETATION AND CONCLUSIONS: This study confirms the possibility of women treated for Hodgkin's disease being able to carry a pregnancy safely to term with the help of steroidal therapy. Careful clinical and obstetric surveillance is important. Focusing attention on long-term survivors of Hodgkin's disease, we set the goal of improving the quality of life of these patients, considering their psychophysical well-being as a whole. Greater attention to the problems of safeguarding fertility in these patients would be advisable, also in the light of legislative regulation of medical care techniques in various countries. PMID- 11418372 TI - A missense mutation (Y1702C) in the coagulation factor V gene is a frequent cause of factor V deficiency in the Italian population. AB - BACKGROUND AND OBJECTIVES: Factor V (FV) deficiency is a rare bleeding disorder whose molecular bases are poorly characterized. We have recently described a FV missense mutation (Y1702C) predicting reduced FV levels in a thrombophilic patient and in a healthy individual. The aim of the present work was to assess the prevalence of the FV Y1702C mutation among subjects with FV deficiency. DESIGN AND METHODS: Carriership of the FV Y1702C mutation was tested in 8 patients with severe FV deficiency (FV:C <8%), in 16 individuals with asymptomatic partial FV deficiency (mean FV:C 38.0%, SD 11.6%) and in 9 patients with pseudo-homozygous APC-resistance (mean FV:C 46.2%, SD 3.6%). An AccI restriction protocol was employed for rapid mutation screening. RESULTS: The FV Y1702C mutation was detected in two unrelated patients with unmeasurable FV levels (one being homozygous and the other doubly heterozygous for a still unknown mutation) and in one subject with partial FV deficiency (FV:C 30%). A striking difference in bleeding phenotype was observed between the homozygous patient and her asymptomatic brother with the same FV genotype. A multi-point FV haplotype analysis was performed in all unrelated carriers of the FV Y1702C mutation. Three haplotypes were found to underlie the mutation in different individuals, suggesting that it might have arisen independently more than once. INTERPRETATION AND CONCLUSIONS: FV Y1702C is a common cause of FV deficiency in the Italian population and might be a recurrent mutation. PMID- 11418373 TI - Inherited thrombophilic risk factors in a large cohort of individuals referred to Italian thrombophilia centers: distinct roles in different clinical settings. AB - BACKGROUND AND OBJECTIVES: Despite inherited thrombophilic risk factors being strongly associated with vein thrombosis, decisions on whether to screen subjects for these factors vary in different clinical settings. DESIGN AND METHODS: We calculated the prevalence of inherited thrombophilic risk factors in a large cohort of patients (n=1,238) with different clinical manifestations of vein thromboembolism. In the present cohort, screening for inherited thrombophilia was worthwhile among patients who developed vein thrombosis of the leg or cerebral vein thrombosis. Carriers of FV Leiden or FII A(20210) mutation more frequently had had deep vein thrombosis of the leg (OR: 4.35; 95% CI: 3.39-5.60), superficial vein thrombosis (OR: 3.34; 95% CI: 2.06-5.41), or cerebral vein thrombosis (OR: 2.77; 95% CI: 1.10-6.96). RESULTS: The screening program appeared to have a limited relevance in patients with isolated pulmonary embolism (OR: 2.13; 95% CI: 1.28-3.54), or mesenteric vein thrombosis (OR: 2.05; 95% CI: 1.22 3.44). INTERPRETATION AND CONCLUSIONS: The lack of association with inherited thrombophilia does not justify routine screening of patients with thrombosis of the upper extremities or with retinal vein thrombosis. PMID- 11418374 TI - Pharmacokinetic evaluation of recombinant, activated factor VII in patients with inherited factor VII deficiency. AB - BACKGROUND AND OBJECTIVES: Recombinant factor VIIa (rFVIIa) has been widely used in the treatment of bleedings occurring in hemophiliacs with inhibitors. Very few reports exist on the use of rFVIIa in patients with inherited FVII deficiency. Pharmacokinetic studies on rFVIIa have been performed exclusively in hemophiliacs, patients with cirrhosis or volunteers pretreated with acenocoumarol. The aim of this study was to evaluate the kinetics of rFVIIa in patients naturally deficient of FVII. DESIGN AND METHODS: A single dose kinetic study with rFVIIa was performed in 5 patients affected by severe congenital deficiency of factor VII in order to evaluate the true kinetic parameters of rFVIIa without the interference of FVII. Two dosages, 15 and 30 microg/kg, were used in a crossover schedule. FVII:C and FVIIa concentration/time curves were analyzed by a model-independent method. Antithrombin (AT), prothombin fragment 1+2 (F1+2) and tissue factor pathway inhibitor (TFPI) were assayed. RESULTS: No differences emerged between the dosages with respect to dose-independent parameters [total body clearance (CL), volume of distribution area (VdArea), mean residence time (MRT)]. No significant changes of AT, TFPI, and F1+2 were observed. Comparing the results with those of other studies performed in adult hemophiliacs, in patients affected by cirrhosis or in volunteers on oral anticoagulant therapy (OAT), CL and VdArea of rFVIIa were definitely higher and in vivo recovery was lower. INTERPRETATION AND CONCLUSIONS: These findings suggest that the kinetics of rFVIIa are not dose-dependent. In the absence of FVII, the changes of VdArea and CL may be in agreement with a mechanism of competition between FVII and rFVIIa for tissue factor binding. PMID- 11418375 TI - Conventional hematopoietic stem cell transplants from identical or alternative donors are feasible in recipients relapsing after an autograft. AB - BACKGROUND AND OBJECTIVES: The risk of relapse after autologous bone marrow transplantation (ASCT) is high and is related to the type of malignancy and phase of the disease. The outcome for the patient who relapses after an autologous transplant is poor. Some of these patients achieve a remission with conventional chemotherapy, but it is usually short-lasting. Most of them succumb to the original disease. One further therapeutic possibility is an allogeneic transplant which would confer the potential advantage of a graft-versus-leukemia effect in addition to the lack of tumor contamination of the graft and to a high-dose intensity conditioning regimen. DESIGN AND METHODS: We have studied the outcome of 31 patients with hematologic malignancies who underwent an allogeneic hematopoietic stem cell transplant (HSCT) after failing an autologous transplant because of relapse (n=29) or persistent aplasia (n=2). The median age at allograft was 36 years (18-55) and the interval from autograft to allograft was 21 months (3-141). The source of stem-cells was unmanipulated bone marrow (n=26) or growth-factor-mobilized peripheral blood (n=5). The donor was an HLA-identical sibling (n=7), or an alternative donor (n=24) (family mismatched n=11, or matched unrelated n=13). The conditioning regimen was cyclophosphamide and thiotepa (n=22), or cyclophosphamide and total body irradiation (n=9) Graft-versus-host disease (GvHD) prophylaxis consisted of cyclosporine (CyA) + methotrexate (MTX). RESULTS: Acute GvHD was scored as 0-I, II, or III-IV in 39%, 48%, and 13% of the patients, respectively. Sixteen patients died of transplant-related complications and one of progressive disease. With a median follow-up of 220 days (9-2104) the actuarial 2-year transplant-related mortality (TRM) was 51%, the actuarial relapse risk 37%, the actuarial survival 46%. Fifteen patients (48%) are alive in complete remission, with a median follow-up of 32 months (range 2-71). INTERPRETATION AND CONCLUSIONS: These data suggest that patients relapsing after an autotransplant should be screened for potential related or unrelated donors: although TRM remains high there is a definite chance of long-term disease-free survival if these patients are allografted. PMID- 11418376 TI - Helper T-lymphocyte precursor frequency predicts the occurrence of graft-versus host disease and disease relapse after allogeneic bone marrow transplantation from HLA-identical siblings. AB - BACKGROUND AND OBJECTIVES: Donor helper T-lymphocytes may be involved in graft versus-host disease (GVHD) and a graft-versus-leukemia effect after bone marrow transplantation (BMT). We assayed donor helper T-lymphocyte precursor frequencies (HTLP(f)) to see whether they could predict the severity of GVHD and disease relapse after transplantation, thereby facilitating donor selection, pre transplant counselling and modification of GVHD prophylaxis after BMT. DESIGN AND METHODS: Thirty-six consecutive adult BMT recipients and their HLA-identical sibling donors were recruited. HTLP((f)) was measured as a function of interleukin-2 secretion by alloreactive donor T-cells using a limiting dilution assay. Patients were followed prospectively to assess the severity of GVHD and the status of the primary disease after BMT. RESULTS: Eight donors had HTLP((f)) less than or equal to 10(-6); no recipients of these grafts developed severe GVHD after transplantation. Twenty-eight donors had HTLP(f) greater than 10(-6) and 18 recipients of these grafts developed severe GVHD (> or = grade 2) (chi(2) test, p<0.01). Seven donors had HTLP(f) greater than 10(-5) and no recipient had disease relapse. Twenty-nine donors had HTLP(f) less than or equal to 10(-5), 11 recipients of these grafts developed disease relapse (chi(2) test, p=0.08). INTERPRETATION AND CONCLUSIONS: BMT recipients from HLA-identical sibling donors with low (<10(-6)) and high (>10(-5)) HTLP(f) may have a low risk of acute GVHD and disease relapse after transplantation. PMID- 11418377 TI - Molecular characterization of thalassemia intermedia with homozygous Hb Malay and Hb Malay/HbE in Thai patients. PMID- 11418378 TI - Metoclopramide-induced methemoglobinemia in a patient with co-existing deficiency of glucose-6-phosphate dehydrogenase and NADH-cytochrome b5 reductase: failure of methylene blue treatment. PMID- 11418379 TI - Elevated plasma thrombopoietin levels and thrombosis in essential thrombocythemia: a preliminary report. PMID- 11418380 TI - Predictive prognostic factors after splenectomy in patients with idiopathic thrombocytopenic purpura. PMID- 11418381 TI - A randomized, prospective comparison of allogeneic bone marrow and peripheral blood progenitor cell transplantation in the treatment of hematologic malignancies: an update. PMID- 11418382 TI - Diminished T-cell recovery after CD34(+) selected autologous peripheral blood stem cell transplantation increases the risk of cytomegalovirus infection. PMID- 11418383 TI - An autopsy-based retrospective study of secondary thrombotic thrombocytopenic purpura. PMID- 11418384 TI - The irreplaceable image: Orbital involvement in multiple myeloma. PMID- 11418385 TI - The irreplaceable image: An unexpected manifestation of anemia. PMID- 11418386 TI - Serious business: radiation safety and radiation protection. PMID- 11418388 TI - Skin injuries from fluoroscopically guided procedures: part 1, characteristics of radiation injury. PMID- 11418389 TI - No-To-Bac. PMID- 11418390 TI - Skin injuries from fluoroscopically guided procedures: part 2, review of 73 cases and recommendations for minimizing dose delivered to patient. PMID- 11418391 TI - Radiation-induced skin injuries and fluoroscopy. PMID- 11418392 TI - Projectile cylinder accidents resulting from the presence of ferromagnetic nitrous oxide or oxygen tanks in the MR suite. AB - OBJECTIVE: The purpose of this study is to alert MR users to the potentially lethal consequences of projectile cylinder accidents in the MR environment. CONCLUSION: Projectile cylinder tank accidents still occur and may be increasing, despite adherence to screening policies before MR imaging and safety education of hospital personnel. Four of the last five accidents at our institutions occurred within the past 3 years. PMID- 11418393 TI - A cost analysis of positron emission tomography. AB - OBJECTIVE: Changes in regulations and improvements in reimbursement have propelled positron emission tomography (PET) into clinical use, making it increasingly important to understand the costs of this emerging service. Cost analyses are important tools to do this. Data published previously on these topics reflect assumptions that are no longer valid. The aim of this study was to determine the cost of developing and operating a PET facility and to evaluate whether a regional cyclotron serving several scanners reduces costs. MATERIALS AND METHODS: Financial data were collected on capital expense and global operating costs through interviews with industry experts, evaluation of prior studies, and review of expenses incurred at the University of Southern California PET center. A data model and cost templates were developed. Expenses were allocated either to the production or purchase of radiopharmaceuticals or to the provision of the PET scan, and the cost per procedure was determined. A sensitivity analysis was performed on the net present value for key parameters. RESULTS: A cyclotron serving a single scanner is not financially viable. The radiopharmaceutical distribution configurations were financially sound. In these cases, the cost of the radiopharmaceutical was approximately $700 per dose with modest levels of production (12 doses per day). In addition, the average cost of PET scans (technical scan and professional charges) ranged from approximately $900 to $1400. The critical factor for profitability was shown to be throughput. CONCLUSION: This analysis provides significant insight into the cost of PET and the comparative costs of offering PET through four operating configurations. Reductions in equipment prices, increased availability of radiopharmaceuticals, growth in demand, and improvements in reimbursement have all contributed to the financial viability of this imaging technique. PMID- 11418394 TI - Patient communication: what to teach radiology residents. PMID- 11418395 TI - Radiology 2000: reinventing radiology--strategies and solutions. PMID- 11418396 TI - Role of abdominal sonography in excluding abdominal malignancy in the initial workup of patients with abdominal complaints. AB - OBJECTIVE: The objective of this study was to evaluate the diagnostic performance and role of abdominal sonography in excluding abdominal malignancy in the initial workup of patients with abdominal complaints. MATERIALS AND METHODS: The sonographic report and follow-up data of 494 patients who had undergone a primary sonographic examination were retrospectively reviewed. Sensitivity and specificity of sonography for the diagnosis of an abdominal malignancy-that is, a primary tumor or metastasis-were determined. Multivariate logistic regression analysis was performed to determine the incremental value of sonography, and a prediction rule was derived. RESULTS: An abnormality on sonography--that is, a mass, ascites, pleural effusion, hydronephrosis, or focal intraparenchymal heterogeneity suggestive of a mass--had a sensitivity for abdominal malignancy of 86% and a specificity of 94%. In the multivariate analysis, the sonographic findings were found to have significant incremental value (odds ratio = 74) after adjustment for the clinical determinants. In patients younger than 38 years with no previous malignancy, no palpable mass, normal liver function test results, and negative findings on sonographic examination, the risk of an abdominal malignancy was less than 1 in 500. CONCLUSION: Our results suggest that sonography may be useful in excluding an abdominal malignancy when used in a primary care setting in patients with abdominal complaints who are at low risk for a malignancy. PMID- 11418397 TI - Sonography of the vertebral arteries: a window to disease of the proximal great vessels. PMID- 11418398 TI - Visualization of areae gastricae on double-contrast upper gastrointestinal radiography: relationship to age of patients. AB - OBJECTIVE: The purpose of this study was to determine whether the frequency of visualization of areae gastricae on double-contrast upper gastrointestinal tract examinations is related to a patient's age. MATERIALS AND METHODS: A total of 141 double-contrast upper gastrointestinal tract examinations with normal findings were reviewed for the presence or absence of areae gastricae on double-contrast images of the stomach. All images were evaluated by two radiologists who were blinded to the age of the patients. The data were then analyzed to determine if the frequency of visualization of areae gastricae on double-contrast studies was significantly related to the age of patients. RESULTS: The frequency of visualization of areae gastricae increased significantly with increasing age (p = 0.008). The youngest age group (20--29 years old) exhibited areae gastricae in only four (19%) of 21 cases, whereas the oldest age group (> or = 70 years old) exhibited areae gastricae in 19 (76%) of 25 cases. On average, the rate of visualization of areae gastricae on double-contrast studies increased by 9% per decade. CONCLUSION: Our data show that the frequency of visualization of areae gastricae on double-contrast upper gastrointestinal tract examinations increases significantly with increasing patient age. It is important for radiologists to be aware of the effect of aging on the delineation of areae gastricae on double contrast studies. PMID- 11418400 TI - Gastrointestinal bezoars: sonographic and CT characteristics. AB - OBJECTIVE: The purpose of this study was to assess the value of imaging studies- conventional abdominal radiographs, sonography, and CT--in the diagnosis of gastrointestinal bezoars. METHODS AND METHODS: A review was made of the radiologic findings of 17 consecutive patients with surgically verified gastrointestinal bezoars over a period of 51 months. RESULTS: Twelve patients had a history of previous gastric surgery. In no patient was a bezoar clinically suspected. Phytobezoars were recorded in 16 patients and a trichobezoar in only one. A total of 33 bezoars were identified at surgery. Two patients had isolated gastric bezoars, whereas 15 patients had bezoars located in the small bowel. Among the latter group, associated gastric bezoars were found in eight patients, and five patients had multiple intestinal bezoars. Abdominal radiographs revealed bezoars in three patients, sonography revealed bezoars in 15, and CT revealed bezoars in all 17. Seven patients had associated gastric bezoars revealed at CT versus only two patients with gastric bezoars revealed at sonography. CT revealed multiple intestinal bezoars in five patients whereas sonography revealed them in only two patients. CONCLUSION: Both sonography and CT are reliable methods for diagnosing gastrointestinal bezoars. CT is more accurate, however, and exhibits a quite characteristic bezoar image; in addition, this imaging technique is able to reveal the presence of additional gastrointestinal bezoars. PMID- 11418401 TI - Lymphoid hyperplasia of the stomach: radiographic findings in five adult patients. AB - OBJECTIVE: The purpose of our study was to report the radiographic findings of biopsy-proven lymphoid hyperplasia of the stomach in five adult patients. CONCLUSION: Lymphoid hyperplasia of the stomach is characterized by distinctive findings on double-contrast upper gastrointestinal tract barium examinations; all five patients had innumerable tiny (1--3 mm in diameter) round frequently umbilicated nodules that carpeted the mucosa of the gastric antrum or antrum and body. Three of these five patients had associated Helicobacter pylori gastritis. The diagnosis of gastric lymphoid hyperplasia, therefore, can be suggested on the basis of the radiographic findings. PMID- 11418402 TI - The Swedish laparoscopic adjustable gastric banding for morbid obesity: radiologic findings in 218 patients. AB - OBJECTIVE: The objective of this study was to determine the prevalence and radiologic features of postoperative complications after Swedish laparoscopic adjustable gastric banding surgery and to emphasize the role of the radiologist in the follow-up of those patients, especially in the treatment of complications. MATERIALS AND METHODS: We reviewed the radiologic findings in 218 consecutive morbidly obese patients after laparoscopic placement of the Swedish gastric banding system. Radiographic studies of the stomach (obtained with liquid barium sulfate suspension) were performed before surgery and 1 month after band placement in every patient. Additional studies in symptomatic patients were performed when needed. RESULTS: Surgical complications found included misplacement of the band (five patients, 2.3%), slippage of the band (17 patients, 7.8%), and pouch enlargement (eight patients, 3.7%). Technical problems encountered were inversion of the access port (three patients, 1.4%), leakage of the device (two patients, 0.9%), and spontaneous decrease of the stoma size caused by gastritis (seven patients, 3.2%) or the hyperosmolar properties of the IV contrast material (12 patients, 5.5%). Intrinsic abnormalities of gastroesophageal tract seen included trapping of food in the stoma (four patients, 1.8%) and esophagitis (11 patients, 5%). CONCLUSION: Although, according to the available data, the gastric banding operation with the Swedish band meets the criteria of a low-risk laparoscopic alternative treatment of morbid obesity, the radiologic appearances of various complications may be seen on the images of patients who have undergone the procedure. The radiologist plays a key role in the early detection of those complications and treatment of specific abnormalities. PMID- 11418403 TI - Killian-Jamieson diverticula: radiographic findings in 16 patients. AB - OBJECTIVE: The purpose of this study was to reassess the findings of Killian Jamieson diverticula (i.e., proximal lateral cervical diverticula) on pharyngoesophagograms and to compare the prevalence, clinical findings, and radiographic findings of Killian-Jamieson diverticula with those of Zenker's diverticulum. MATERIALS AND METHODS: A computerized search of radiology files revealed 16 patients with Killian-Jamieson diverticula and 26 patients with a Zenker's diverticulum. The double-contrast pharyngoesophagograms and medical records were reviewed retrospectively. RESULTS: Only three (19%) of 16 patients with Killian-Jamieson diverticula had symptoms attributable to the diverticula (suprasternal dysphagia in two and cough in one), and none had aspiration pneumonia. In contrast, 16 (62%) of 26 patients with a Zenker's diverticulum had suprasternal dysphagia and three patients (12%) had aspiration pneumonia. Twenty Killian-Jamieson diverticula were detected on pharyngoesophagograms in 16 patients, including 12 (75%) with unilateral left-sided diverticula and four (25%) with bilateral diverticula. The Killian-Jamieson diverticula had an average maximal dimension of 1.4 cm. Zenker's diverticulum was nearly four times as common as Killian-Jamieson diverticula and had an average maximal dimension of 2.5 cm. Three patients (11%) with a Zenker's diverticulum had reflux of barium from the diverticula into the hypopharynx with overflow aspiration. Finally, gastroesophageal reflux was detected in nearly twice as many patients with a Zenker's diverticulum as with Killian-Jamieson diverticula. CONCLUSION: Killian Jamieson diverticula are less common and smaller than Zenker's diverticulum. Killian-Jamieson diverticula are less likely to cause symptoms and are less likely to be associated with overflow aspiration or gastroesophageal reflux than is Zenker's diverticulum. PMID- 11418404 TI - CT appearance of some colonic villous tumors. AB - OBJECTIVE: A review was made of the CT studies and pathology reports of four patients with surgically resected colonic villous adenomatous tumors, two of whom had focal carcinomatous invasion. CONCLUSION: Two patients had villous tumors with IV contrast-enhancing convolutional gyral patterns. The other two patients had tumor masses that showed oral contrast medium collecting in surface interstices, analogous to findings with barium enemas. One of the latter also had an unusual cluster of mesenteric vessels adjacent to the lesion. PMID- 11418405 TI - CT of cecal volvulus: unraveling the image. PMID- 11418406 TI - Hydrocolonic sonography for evaluating inflammatory bowel disease. AB - OBJECTIVE: The purpose of our study was to compare the usefulness of hydrocolonic sonography and (99m)Tc-hexamethylpropyleneamine oxime (HMPAO)--labeled leukocyte scintigraphy in the examination of patients with inflammatory bowel disease, using precise sonographic criteria of bowel involvement. SUBJECTS AND METHODS: Sixty-eight consecutive patients with active inflammatory bowel disease (34 ulcerative colitis and 34 Crohn's disease), 12 with inactive inflammatory bowel disease, and 10 control subjects were prospectively studied. Patients with active disease underwent clinical assessment, hydrocolonic sonography, scintigraphy, and colonoscopy within 72 hr, whereas patients with inactive disease and control subjects underwent clinical examination and hydrocolonic sonography. RESULTS: Involvement of a colonic segment by active inflammatory bowel disease was best defined by mucosal thickness greater than 1.5 mm, bowel wall thickness greater than 4 mm, mucosal irregularity, or the absence of haustra; and involvement of the terminal ileum by bowel wall thickness greater than 4 mm. Using these criteria, hydrocolonic sonography had 100% sensitivity for identifying patients with active inflammatory bowel disease and a greater overall accuracy (87%) than scintigraphy (77%) in the assessment of disease extension. In addition, strong correlation was shown between a hydrocolonic sonography activity index and clinical and endoscopic activity indexes. CONCLUSION: This prospective study provides precise sonographic criteria for the definition of bowel involvement by active inflammatory bowel disease. Hydrocolonic sonography has a greater accuracy than scintigraphy for assessing disease extension and activity. Therefore, hydrocolonic sonography should be considered a first-choice technique to complete the study of inflammatory bowel disease after confirmation of the diagnosis by histology. PMID- 11418407 TI - Evaluation of pelvic adhesions using multiphase and multislice MR imaging with kinematic display. AB - OBJECTIVE: The purpose of this study was to evaluate whether kinematic multiphase and multislice MR imaging could reveal pelvic adhesions. SUBJECTS AND METHODS: Before surgery, 52 women with gynecologic disorders underwent half-Fourier acquisition single-shot fast spin-echo imaging with multiphase and multislice acquisitions. Images were displayed in a cine mode, and the motion of each organ against adjacent organs was evaluated by two radiologists who were unaware of the patients' histories or of the findings of their clinical examinations or surgeries. Findings from MR imaging were classified into three types relative to the adjacent organs: type 1, sliding, defined as organs moving 1 cm or more; type 2, fine motion, defined as organs moving less than 1 cm; or type 3, no motion. Type 2 was further subdivided into two groups: type 2-A, independent fine movement, and type 2-B, synchronous fine movement. All MR imaging findings were verified by laparotomy or laparoscopy. Peristalsis at the rectum, colon, and small intestine were also evaluated. RESULTS: A total of 317 interfaces were evaluated. For findings of type 1 (n = 8 interfaces) and type 2-A (n = 245) on kinematic MR imaging, the negative predictive values for adhesions were 100% and 95.5%, respectively. Findings of type 2-B (n = 52) and type 3 (n = 12) with no adhesions were observed in 40.4% and 66.7%, respectively. When type 1 and type 2 A were regarded as negative findings of adhesions, and type 2-B and type 3 as positive, sensitivity, specificity, and accuracy were 72.5%, 87.4%, and 85.4%, respectively. Peristalsis was observed in 69.2% of patients at the rectum, 86.5% at the colon, and 100% at the small intestine. CONCLUSION: Multiphase and multislice MR imaging with kinematic display may provide new information about the presence of pelvic adhesions. PMID- 11418408 TI - MR imaging of intrahepatic cholangiocarcinoma: use of ferumoxides for lesion localization and extension. AB - OBJECTIVE: The purpose of this study was to evaluate the use of ferumoxides in the visualization and localization of intrahepatic cholangiocarcinoma. CONCLUSION: The contrast-to-noise ratio of cholangiocarcinoma compared with that of the adjacent liver significantly (p < 0.03) improves after ferumoxides administration. Ferumoxides-enhanced MR imaging is a useful technique for the visualization and localization of intrahepatic cholangiocarcinoma. PMID- 11418409 TI - Value of intraarterial prostaglandin E(1) injection during CT hepatic arteriography. AB - OBJECTIVE: The purpose of our investigation was to determine if injection of prostaglandin E(1) during CT hepatic arteriography could help physicians to distinguish tumors from nonportal venous flow-related pseudolesions in the region of the gallbladder fossa. SUBJECTS AND METHODS: In 34 patients who underwent CT during arterial portography to detect liver tumors, CT hepatic arteriography was performed before and after prostaglandin E(1) injection via the superior mesenteric artery. Between each study, an interval of 10 minutes was set. On CT hepatic arteriogram obtained 15 to 20 sec after prostaglandin E(1) injection, we distinguished changes in the size and shape of pseudolesions in the liver around the gallbladder as well as those of 42 tumorous lesions. In addition, we measured the change in CT attenuation of pseudolesions. RESULTS: The size of the enhanced area of pseudolesions visible on CT hepatic arteriography decreased in 69% (25/36) of the pseudolesions after intraarterial prostaglandin E(1) injection, with the mean diameter diminishing from 14.1 mm to 8.8 mm. Notably, in 11 pseudolesions, the enhanced area disappeared. In 86% (31/36), the CT attenuation decreased with the mean attenuation, diminishing from 211.3 H to 163.8 H. However, the size and shape of the enhanced area of tumorous lesions did not change. CONCLUSION: The hemodynamic features of pseudolesions on angiographically assisted helical CT scans caused by cholecystic venous inflow are easily influenced by increased portal venous flow. Consequently, pseudolesions around the gallbladder usually can be distinguished from tumorous lesions by adding prostaglandin E(1) injection via the superior mesenteric artery during CT hepatic arteriography. PMID- 11418410 TI - Agenesis of the hepatic segment of the inferior vena cava with portal continuation. PMID- 11418411 TI - Preoperative diagnosis of ovarian tumors with MR imaging: comparison with transvaginal sonography, positron emission tomography, and histologic findings. AB - OBJECTIVE: Our study evaluated the diagnostic performance of MR imaging compared with that of transvaginal sonography and positron emission tomography (PET) in patients with clinically asymptomatic adnexal findings. An additional goal was to determine whether the combination of the three methods enhanced their diagnostic accuracy. SUBJECTS AND METHODS: Included in the study were 103 women with suspicious adnexal findings on sonography. Patients underwent transvaginal sonography, MR imaging, and PET within 3 weeks of the initial sonography. For MR imaging, axial and sagittal T1-weighted gradient-echo sequences (unenhanced and enhanced) and T2-weighted turbo-spin-echo sequences were acquired. Transvaginal sonography was performed with a 7.5-MHz transducer head. For PET, a modern full ring scanner was used. The results of diagnostic imaging techniques were first evaluated separately, and reviewers were blinded to the results of other methods. Finally, a second session resulted in a consensus diagnosis based on the findings of all three methods. Results of histology were considered the gold standard. RESULTS: Histology revealed 12 malignant and 91 benign ovarian tumors. The following data were calculated for MR imaging, transvaginal sonography, PET, and consensus diagnosis: sensitivities, 83%, 92%, 58%, 92%; specificities, 84%, 59%, 78%, 84%; diagnostic accuracies, 83%, 63%, 76%, 85%, respectively. MR imaging, particularly with contrast-enhanced fat-saturated T1-weighted sequences, was found to correctly reveal dermoid and endometrial cysts. All three methods had false-negative findings with borderline tumors. CONCLUSION: Transvaginal sonography is the diagnostic method of choice as a screening technique for ovarian processes. Suspicious findings on transvaginal sonography should be confirmed on MR imaging. If MR imaging confirms a dermoid or endometrial cyst, further diagnostic procedures may be unnecessary. In all other cases, a surgical evaluation must be considered. PMID- 11418412 TI - Ovarian carcinosarcoma [correction of carcinoma]. PMID- 11418413 TI - Hysterosalpingography: spectrum of normal variants and nonpathologic findings. PMID- 11418414 TI - Successful evaluation of pseudoaneurysm formation after blunt renal injury with dual-phase contrast-enhanced helical CT. PMID- 11418415 TI - Transvaginal catheter drainage of tuboovarian abscess using the trocar method: technique and literature review. PMID- 11418416 TI - Is selective embolization of uterine arteries a safe alternative to hysterectomy in patients with postpartum hemorrhage? AB - OBJECTIVE: The purpose of this study was to evaluate the efficacy and safety of selective arterial embolization to control severe postpartum hemorrhage. MATERIALS AND METHODS: Twenty-five women with intractable postpartum hemorrhage underwent uterine embolization in our institution during a 6-year period. RESULTS: Angiography revealed arterial extravasation in 13 patients (52%). Sixty nine arteries were embolized. External bleeding resolved immediately or was markedly decreased in 24 women. In one patient, embolization failed to control the bleeding, and surgical treatment was required. No major complication of embolization therapy was observed. Ten women were followed up for an average of 2 years. Menstruation resumed in all patients, and one woman became pregnant. CONCLUSION: Embolization of acute postpartum hemorrhage is a safe and effective alternative to hysterectomy. PMID- 11418417 TI - Perfusion-modulated MR imaging-guided radiofrequency ablation of the kidney in a porcine model. AB - OBJECTIVE: This study was performed to test the hypothesis that temporary renal ischemia will result in increased thermal lesion size during radiofrequency thermal ablation in the kidney. MATERIALS AND METHODS: Twelve kidneys were treated in six pigs that were placed under general anesthesia in the MR suite, using a 0.2-T open C-shaped MR imaging system. A 4-cm-long, 14-mm-diameter balloon catheter was placed into the aorta using a transfemoral approach, and the balloon was positioned proximal to the renal arteries via guidance with MR imaging. A 2-cm exposed-tip MR-compatible 17-gauge radiofrequency electrode was placed into one kidney under MR fluoroscopy using fast imaging with steady-state free precession (FISP) sequences. Thermal ablation was performed with the electrode tip temperature maintained at 90 +/- 2 degrees C for 10 min. This procedure was repeated in the contralateral kidney. The balloon was inflated during one ablation. Postablation images were obtained, the pigs were sacrificed, and both kidneys of each animal were harvested for pathologic correlation. RESULTS: Technical success was achieved in all animals. The lesion measured 14.2 +/- 2.2 mm (mean +/- standard deviation) for the ischemic kidney versus 8.0 +/- 2.6 mm in the normally perfused kidney (p = 0.00002). No significant complications were noted. In all images, thermal lesions displayed low signal intensity with a sharp rim of high signal intensity best visualized using short tau inversion recovery (STIR) sequences with a mean accuracy of 1.3 +/- 1.2 mm when compared with pathologic findings and a mean contrast-to-noise ratio of 4.9 +/- 2.5. CONCLUSION: We accept the hypothesis that temporary renal ischemia leads to a significantly increased radiofrequency ablation lesion size. We conclude that catheter-based balloon perfusion reduction is feasible, that the procedure does not lead to major complications, and that it can be performed using MR imaging as the sole imaging modality. PMID- 11418418 TI - MR imaging-guided biopsy and therapeutic intervention in a closed-configuration magnet: single-center series of 361 punctures. AB - OBJECTIVE: The purpose of the study was to evaluate practical experience with interventional MR imaging in a closed-bore 1.5-T imaging system. A total of 361 MR-guided biopsies were performed in 250 patients and analyzed retrospectively. The technique comprised four steps: localization; planning; action (cutting or aspiration biopsy, or instillation of a therapeutic agent), with verification in two perpendicular planes; and obtaining control scans. CONCLUSION: The mean duration of a biopsy was 21 min; there were no major complications. Image contrast, signal, matrix options, and visibility of needle track and tip position permitted uncomplicated orientation. The interventional MR technique could be applied in any puncture setting. PMID- 11418420 TI - Calcifications highly suggestive of malignancy: comparison of breast biopsy methods. AB - OBJECTIVE: The purpose of this study was to compare the usefulness of, and cost of diagnosing with, different breast biopsy methods for women with calcifications highly suggestive of malignancy. MATERIALS AND METHODS: One hundred thirty-nine women with calcifications highly suggestive of malignancy underwent diagnostic biopsy. Of these, 89 women had stereotactic biopsy with a 14-gauge automated needle (n = 25), 14-gauge vacuum-assisted probe (n = 17), or 11-gauge vacuum assisted probe (n = 47); and 50 women had diagnostic surgical biopsy. Medical records were reviewed. Cost savings for stereotactic biopsy were calculated using Medicare data. RESULTS: The median number of operations was one for women who had stereotactic biopsy versus two for women who had diagnostic surgical biopsy. The likelihood of undergoing a single operation was significantly greater for women who had stereotactic rather than surgical biopsy, among all women (61/89 [68.5%] vs. 19/50 [38.0%], p < 0.001) and among women treated for breast cancer (55/77 [71.4%] vs. 6/37 [16.2%], p = 0.0000001). Stereotactic 11-gauge vacuum-assisted biopsy, as compared with 14-gauge automated core or 14-gauge vacuum-assisted biopsy, was significantly more likely to spare a surgical procedure (36/47 [76.6%] vs. 16/42 [38.1%], p = 0.0005). Stereotactic 11-gauge vacuum-assisted biopsy resulted in the greatest cost reduction, yielding savings of $315 per case compared with diagnostic surgical biopsy; for women with solitary lesions, stereotactic 11-gauge biopsy decreased the cost of diagnosis by 22.2% ($334/$1502). CONCLUSION: For women with calcifications highly suggestive of malignancy, the use of stereotactic rather than surgical biopsy decreases the number of operations. Stereotactic 11-gauge vacuum-assisted biopsy, as compared with 14-gauge automated core or 14-gauge vacuum-assisted biopsy, is significantly more likely to spare a surgical procedure and has the highest cost savings. PMID- 11418421 TI - The pendent view: an additional projection to confirm the diagnosis of milk of calcium. PMID- 11418422 TI - Primary breast lymphoma originating in a benign intramammary lymph node. PMID- 11418423 TI - Multislice helical CT of focal and diffuse lung disease: comprehensive diagnosis with reconstruction of contiguous and high-resolution CT sections from a single thin-collimation scan. AB - OBJECTIVE: We tested breath-held 1-mm multislice helical CT for obtaining both contiguous and high-resolution CT sections of the chest from a single set of raw data. SUBJECTS AND METHODS: Seventy patients with suspected focal and diffuse lung disease were allocated into two groups for comparison. The first group (n = 35) underwent multislice helical CT of the chest with 1-mm collimation and a pitch of 6. From the raw data, 5-mm contiguous and 1.25-mm high-resolution CT sections were reconstructed. The second group (n = 35) underwent conventional single-slice helical CT and high-resolution CT. High-resolution CT sections and 5 mm scans were rated for overall image quality, spatial resolution, subjective signal-to-noise ratio, diagnostic value, depiction of bronchi and parenchyma, and motion and streak artifacts. The 5-mm scans were also rated for contrast resolution and depiction of the heart and vessels. Radiation dose was calculated. RESULTS: We rated 5-mm multislice helical CT superior to 5-mm single-slice helical CT, having a significantly higher total score (p = 0.0001). No significant difference (p = 0.986) was found between multislice and single-slice high-resolution CT sections. Radiation dose was 5.55 mSv for multislice helical CT and 5.50 mSv for single-slice helical CT. CONCLUSION: Contiguous chest scans of superior quality and high-resolution CT sections of equal image quality compared with single-slice helical CT can be obtained using multislice helical CT. Therefore, a comprehensive diagnosis is feasible in patients with suspected focal and diffuse lung disease by obtaining a single scan. PMID- 11418424 TI - Oxygen-enhanced MR ventilation imaging of the lung: preliminary clinical experience in 25 subjects. AB - OBJECTIVE: The purpose of this study was to show the feasibility of oxygen enhanced MR ventilation imaging in a clinical setting with correlation to standard pulmonary function tests, high-resolution CT, and (81m)Kr ventilation scintigraphy. SUBJECTS AND METHODS: Seven healthy volunteers, 10 lung cancer patients, and eight lung cancer patients with pulmonary emphysema were studied. A respiratory synchronized inversion-recovery single-shot turbo-spin-echo sequence (TE, 16; inversion time, 720 msec; interecho spacing, 4 msec) was used for data acquisition. The following paradigm of oxygen inhalation was used: 21% oxygen (room air), 100% oxygen, 21% oxygen. MR imaging data including maximum mean relative enhancement ratio and mean slope of relative enhancement were correlated with forced expiratory volume in 1 sec, diffusing lung capacity, high-resolution CT emphysema score, and mean distribution ratio of (81m)Kr ventilation scintigraphy. RESULTS: Oxygen-enhanced MR ventilation images were obtained in all subjects. Maximum mean relative enhancement ratio and mean slope of relative enhancement of lung cancer patients were significantly decreased compared with those of the healthy volunteers (p < 0.0001, p < 0.0001). The mean slope of relative enhancement in lung cancer patients with pulmonary emphysema was significantly lower than that of lung cancer patients without pulmonary emphysema (p < 0.0001). Maximum mean relative enhancement ratio (r(2) = 0.81) was excellently correlated with diffusing lung capacity. Mean slope of relative enhancement (r(2) = 0.74) was strongly correlated with forced expiratory volume in 1 sec. Maximum mean relative enhancement had good correlation with the high resolution CT emphysema score (r(2) = 0.38). The maximum mean relative enhancement had a strong correlation with the distribution ratio (r(2) = 0.77). CONCLUSION: Oxygen-enhanced MR ventilation imaging in human subjects showed regional changes in ventilation, thus reflecting regional lung function. PMID- 11418425 TI - Paddle-wheel CT display of pulmonary arteries and other lung structures: a new imaging approach. PMID- 11418426 TI - Sarcoidosis induced by interferon therapy. PMID- 11418427 TI - Pericardial varices: depiction on three-dimensional CT angiography. PMID- 11418428 TI - Endomyocardial fibrosis mimicking right ventricular tumor. PMID- 11418429 TI - Aortic dissection: CT features that distinguish true lumen from false lumen. AB - OBJECTIVE: The purpose of this study was to determine which CT findings are reliable indicators of the true or false lumen in an aortic dissection. CONCLUSION: The beak sign and a larger cross-sectional area were the most useful indicators of the false lumen for both acute and chronic dissections. Features generally indicative of the true lumen included outer wall calcification and eccentric flap calcification. In cases showing one lumen wrapping around the other lumen in the aortic arch, the inner lumen was invariably the true lumen. PMID- 11418431 TI - Diagnostic disagreement of imaging quantitative sonography of the calcaneus with dual X-ray absorptiometry of the spine and femur. AB - OBJECTIVE: This study evaluates the diagnostic agreement between imaging quantitative sonography of the calcaneus and dual X-ray absorptiometry of the spine and femur for diagnosing osteoporosis. MATERIALS AND METHODS: In 498 female patients (56 +/- 18 years old), bone mineral density measurements by dual X-ray absorptiometry of the lumbar spine (posteroanterior, L1--L4) and the proximal femur and imaging quantitative sonography of the calcaneus were performed. The percentage of patients having T-scores less than or equal to a threshold of -2.5 standard deviations below a young normal reference was used to compare quantitative sonography with dual X-ray absorptiometry. The diagnostic agreement was assessed using kappa scores. RESULTS: Approximately 30% of the patients had a T-score less than or equal to -2.5 standard deviations as assessed by imaging quantitative sonography (broadband ultrasound attenuation), 26.5% as assessed by dual X-ray absorptiometry of the spine, and 16.7--56.4% as assessed by dual X-ray absorptiometry of the different regions of interest at the femur. Kappa analysis showed that severe diagnostic disagreement exists among broadband ultrasound attenuation and dual X-ray absorptiometry (kappa = 0.28-0.42). CONCLUSION: Considerable diagnostic disagreement exists between imaging quantitative sonography and dual X-ray absorptiometry of the spine and femur. The disagreement is in the same range as that reported recently in comparisons of dual X-ray absorptiometry and nonimaging quantitative sonography. In general, no distinct advantage for imaging quantitative sonography could be found when compared with other techniques. PMID- 11418432 TI - MR arthrography of the glenohumeral joint: a tailored approach. AB - OBJECTIVE: The purpose of this study was to anatomically confirm that anterior shoulder injection could result in penetration of the anterior stabilizing structures of the glenohumeral joint and to advocate the use of a tailored approach to MR arthrography based on presenting symptoms. CONCLUSION: A tailored approach to MR arthrography may be a useful way to isolate expected pathology in the shoulder and limit confounding findings related to the performance of the procedure. PMID- 11418433 TI - Synovial plicae in the knee. PMID- 11418435 TI - Whole-body MR imaging for detection of bone metastases in children and young adults: comparison with skeletal scintigraphy and FDG PET. AB - OBJECTIVE: The purpose of this study was to compare the diagnostic accuracy of whole-body MR imaging, skeletal scintigraphy, and 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) for the detection of bone metastases in children. SUBJECTS AND METHODS: Thirty-nine children and young adults who were 2- 19 years old and who had Ewing's sarcoma, osteosarcoma, lymphoma, rhabdomyosarcoma, melanoma, and Langerhans' cell histiocytosis underwent whole body spin-echo MR imaging, skeletal scintigraphy, and FDG PET for the initial staging of bone marrow metastases. The number and location of bone and bone marrow lesions diagnosed with each imaging modality were correlated with biopsy and clinical follow-up as the standard of reference. RESULTS: Twenty-one patients exhibited 51 bone metastases. Sensitivities for the detection of bone metastases were 90% for FDG PET, 82% for whole-body MR imaging, and 71% for skeletal scintigraphy; these data were significantly different (p < 0.05). False-negative lesions were different for the three imaging modalities, mainly depending on lesion location. Most false-positive lesions were diagnosed using FDG PET. CONCLUSION: Whole-body MR imaging has a higher sensitivity than skeletal scintigraphy for the detection of bone marrow metastases but a lower sensitivity than FDG PET. PMID- 11418436 TI - CT detection of mandibular invasion by squamous cell carcinoma of the oral cavity. AB - OBJECTIVE: The purpose of this study was to determine the diagnostic accuracy of CT in detecting mandibular invasion by squamous cell carcinoma of the oral cavity. MATERIALS AND METHODS: Forty-nine patients who had squamous cell carcinoma of the oral cavity that was clinically fixed to the mandible were treated with mandibulectomy. All patients underwent contrast-enhanced CT (contiguous 3-mm-thick sections) through the primary site before surgery. All studies were reconstructed with bone algorithm. These studies were retrospectively reviewed by a neuroradiologist for evidence of mandibular invasion. The imaging results were compared with the histologic findings in all cases. RESULTS: CT correctly revealed 25 of 26 cases with mandibular invasion. CT correctly excluded mandibular invasion in 20 of 23 cases without invasion. The diagnostic accuracy of CT for detecting mandibular invasion was as follows: sensitivity, 96%; specificity, 87%; positive predictive value, 89%; and negative predictive value, 95%. CONCLUSION: Thin-section (3-mm) CT reconstructed with bone algorithm is an accurate technique to detect mandibular involvement by squamous cell carcinoma of the oral cavity. PMID- 11418437 TI - Late pulmonary metastases from hemangiopericytoma of the mandible: unusual findings on CT and MR imaging. PMID- 11418438 TI - Writing, signing, and reading the radiology report: who is responsible and when? PMID- 11418439 TI - Radiofrequency ablation of spinal tumors. PMID- 11418440 TI - Communication, the deep pocket, and ACR standards. PMID- 11418441 TI - Less is not more in stereotactic biopsy. PMID- 11418442 TI - Fat collection related to the intrahepatic portion of the inferior vena cava. PMID- 11418443 TI - Satisfaction of search in osteoradiology. PMID- 11418444 TI - Transient aortic thrombus. PMID- 11418445 TI - Heeding the call: radiologists in the emergency department. PMID- 11418447 TI - Why was Emergency Radiology not cited? PMID- 11418448 TI - Small-bowel obstruction due to migratory trichobezoar in a child: CT diagnosis. PMID- 11418449 TI - Mycotic pseudoaneurysm presenting as a pulsatile mass at sternotomy incision. PMID- 11418450 TI - Os peroneal fracture with associated peroneus longus tendinopathy. PMID- 11418451 TI - Mesalamine-induced unilateral eosinophilic pneumonia. PMID- 11418453 TI - Introduction: spatial origin of murine hematopoietic stem cells. PMID- 11418454 TI - Generation of definitive hematopoietic stem cells from murine early yolk sac and paraaortic splanchnopleures by aorta-gonad-mesonephros region-derived stromal cells. AB - There is controversy as to whether murine definitive hematopoiesis originates from yolk sac (YS) or the intraembryonic region. This study reports the generation of definitive hematopoietic stem cells (HSCs) from both early YS and intraembryonic paraaortic splanchnopleures (P-Sp) on AGM-S3 stromal cells derived from the aorta-gonad-mesonephros (AGM) region at 10.5 days post coitum (dpc). YS and P-Sp cells at 8.5 dpc generated no definitive hematopoiesis-derived colony forming cells in cocultures with AGM-S3 cells, but spleen colony-forming cells and HSCs capable of reconstituting definitive hematopoiesis in adult mice simultaneously appeared on day 4 of coculture. Precursors for definitive HSCs were present in YS and P-Sp at 8.0 dpc, a time when YS and embryo were not connected by blood vessels. It is proposed that precursors with the potential to generate definitive HSCs appear independently in YS and intraembryonic P-Sp and that the P-Sp or AGM region affords the microenvironment that facilitates generation of definitive hematopoiesis from precursors. PMID- 11418455 TI - Structure of a factor VIII C2 domain-immunoglobulin G4kappa Fab complex: identification of an inhibitory antibody epitope on the surface of factor VIII. AB - The development of an immune response to infused factor VIII is a complication affecting many patients with hemophilia A. Inhibitor antibodies bind to antigenic determinants on the factor VIII molecule and block its procoagulant activity. A patient-derived inhibitory immunoglobulin G4kappa antibody (BO2C11) produced by an immortalized memory B-lymphocyte cell line interferes with the binding of factor VIII to phospholipid surfaces and to von Willebrand factor. The structure of a Fab fragment derived from this antibody complexed with the factor VIII C2 domain was determined at 2.0 A resolution. The Fab interacts with solvent-exposed basic and hydrophobic side chains that form a membrane-association surface of factor VIII. This atomic resolution structure suggests a variety of amino acid substitutions in the C2 domain of factor VIII that might prevent the binding of anti-C2 inhibitor antibodies without significantly compromising the procoagulant functions of factor VIII. PMID- 11418456 TI - Genetic hypercoagulability: screening should be an informed choice. PMID- 11418457 TI - Genetic hypercoagulability: prevention suggests testing family members. PMID- 11418458 TI - Blood levels of immune cells predict survival in myeloma patients: results of an Eastern Cooperative Oncology Group phase 3 trial for newly diagnosed multiple myeloma patients. AB - Previously, it was reported that patients with multiple myeloma (MM) who have higher baseline levels of blood CD4(+) or CD19(+) cells have longer survival. This article extends the analysis of immune cell levels and survival in a large cohort (N = 504) of patients with MM entered on Eastern Cooperative Oncology Group (ECOG) phase 3 trial (9486). Newly diagnosed patients with MM received 2 cycles of vincristine, bischloroethylnitrosourea, melphalan, cytoxan, prednisone (VBMCP) and were treated on one of 3 randomized arms: VBMCP with either interferon or high-dose cyclophosphamide, or VBMCP alone. Blood immune cell levels were studied at trial entry (baseline), after 2 cycles of chemotherapy, after 2 years of therapy, and at relapse. Baseline CD3(+), CD4(+), CD8(+), CD19(+), and CD4(+) subset cell levels were all positively associated with survival (P =.0087 to P <.0001). A multivariate analysis incorporating CD4(+) and CD19(+) cell levels defined 3 separate groups of patients with MM to survival outcome. Higher CD19(+) blood levels were positively associated with MM-patient survival at entry to the study, at year 2, and at relapse (P <.0001 at all 3 timepoints). Patients with MM had evidence of immune cell reconstitution after 2 years of therapy, but the rate and extent of recovery was greater for CD8(+), which was greater than CD4(+), which was greater than CD19(+). This latter data affirms the positive relationship between the quantitative status of the blood immune system in MM and survival. In addition, the importance of the CD19(+) blood cells to survival is evident throughout the course of MM. Therapeutic efforts to maintain an intact immune system may be crucial in maximizing chemotherapeutic and/or immunotherapy efforts in this disease. PMID- 11418459 TI - Quantitation of minimal disease levels in chronic lymphocytic leukemia using a sensitive flow cytometric assay improves the prediction of outcome and can be used to optimize therapy. AB - Previous studies have suggested that the level of residual disease at the end of therapy predicts outcome in chronic lymphocytic leukemia (CLL). However, available methods for detecting CLL cells are either insensitive or not routinely applicable. A flow cytometric assay was developed that can differentiate CLL cells from normal B cells on the basis of their CD19/CD5/CD20/CD79b expression. The assay is rapid and can detect one CLL cell in 10(4) to 10(5) leukocytes in all patients. We have compared this assay to conventional assessment in 104 patients treated with CAMPATH-1H and/or autologous transplant. During CAMPATH-1H therapy, circulating CLL cells were rapidly depleted in responding patients, but remained detectable in nonresponders. Patients with more than 0.01 x 10(9)/L circulating CLL cells always had significant (> 5%) marrow disease, and blood monitoring could be used to time marrow assessments. In 25 out of 104 patients achieving complete remission by National Cancer Institute (NCI) criteria, the detection of residual bone marrow disease at more than 0.05% of leukocytes in 6 out of 25 patients predicted significantly poorer event-free (P =.0001) and overall survival (P =.007). CLL cells are detectable at a median of 15.8 months (range, 5.5-41.8) posttreatment in 9 out of 18 evaluable patients with less than 0.05% CLL cells at end of treatment. All patients with detectable disease have progressively increasing disease levels on follow-up. The use of sensitive techniques, such as the flow assay described here, allow accurate quantitation of disease levels and provide an accurate method for guiding therapy and predicting outcome. These results suggest that the eradication of detectable disease may lead to improved survival and should be tested in future studies. PMID- 11418460 TI - Platelet glycoprotein Ibalpha Kozak polymorphism is associated with an increased risk of ischemic stroke. AB - Platelets are pivotal to the process of arterial thrombosis resulting in ischemic stroke. Occlusive thrombosis is initiated by the interaction of von Willebrand factor (vWf) and platelet glycoprotein (GP) Ibalpha. Three polymorphisms have been described in GP Ibalpha (Kozak T/C polymorphism, variable number of tandem repeats [VNTR], and the human platelet antigen 2a [HPA-2a] [Thr] or HPA-2b [Met] at position 145), each of which may enhance the vWf and GP Ibalpha interaction. This study investigated whether these polymorphisms are candidate genes for first ever ischemic stroke. A hospital-based case-control study was conducted of 219 cases of first-ever ischemic stroke and 205 community controls randomly selected from the electoral roll and stratified by age, sex, and postal code. The subtypes of stroke were classified, the prevalence of conventional risk factors was recorded, and blood was collected to perform genotyping analysis for Kozak C or T alleles, VNTR, and HPA-2a/b. It was found that the Kozak T/C genotype was over represented in the stroke group (32.2%) compared with controls (22.8%) (odds ratio [OR], 1.6; 95% confidence interval [CI], 1.03-2.54; P <.03), and the association was still present even after adjusting for conventional risk factors. There was a trend in the increased prevalence of HPA-2a/b in stroke patients (15%) compared with controls (9.9%) (adjusted OR, 1.8; 95% CI, 0.94-3.4; P =.07). No associations were seen with the VNTR polymorphism or with any of the polymorphisms with stroke subtype. It was concluded that the Kozak T/C polymorphism, which is associated with an increase in platelet GP Ibalpha surface expression, is an independent risk factor for first-ever ischemic stroke. PMID- 11418461 TI - Prognostic factors and response to fludarabine therapy in patients with Waldenstrom macroglobulinemia: results of United States intergroup trial (Southwest Oncology Group S9003). AB - Current information on Waldenstrom macroglobulinemia (WM) is based on retrospective or single-institution studies of patients requiring therapy. Between 1992 and 1998, 231 patients with WM were enrolled in a prospective observational multicenter clinical trial. Of these, 182 patients with symptomatic or progressive disease were treated with 4 to 8 cycles of therapy with a purine nucleoside analogue, fludarabine (FAMP; 30 mg/m(2) of body-surface area daily for 5 days every 28 days). A serum beta2-microglobulin (beta2M) level below 3 mg/L and a hemoglobin level of at least 120 g/L (12 g/dL) at presentation predicted a lower likelihood of requiring therapy. The overall rate of response to FAMP therapy was 36% (95% confidence interval, 29%-44%), with 2% complete remissions. Patients who were 70 years old or older had a substantially lower likelihood of response (odds ratio, 0.34; P =.004) than younger patients. On multivariate analysis, a serum beta2M level of 3 mg/L or higher, hemoglobin level below 120 g/L, and serum IgM level below 40 g/L [4 g/dL] were significant adverse prognostic factors for survival. We developed a simple staging system for WM by using these variables and identified 4 distinct subsets of patients with estimated 5-year overall survival rates of 87%, 64%, 53%, and 22%, and 5-year progression-free survival rates of 83%, 55%, 33%, and 12%. Prognosis in WM is highly variable and serum beta2M was the dominant predictor of a need for therapy and of survival. FAMP has activity against WM. Our staging system may provide guidance for a risk-based approach to the treatment of WM. PMID- 11418462 TI - Highly efficient gene delivery by mRNA electroporation in human hematopoietic cells: superiority to lipofection and passive pulsing of mRNA and to electroporation of plasmid cDNA for tumor antigen loading of dendritic cells. AB - Designing effective strategies to load human dendritic cells (DCs) with tumor antigens is a challenging approach for DC-based tumor vaccines. Here, a cytoplasmic expression system based on mRNA electroporation to efficiently introduce tumor antigens into DCs is described. Preliminary experiments in K562 cells using an enhanced green fluorescent protein (EGFP) reporter gene revealed that mRNA electroporation as compared with plasmid DNA electroporation showed a markedly improved transfection efficiency (89% versus 40% EGFP(+) cells, respectively) and induced a strikingly lower cell toxicity (15% death rate with mRNA versus 51% with plasmid DNA). Next, mRNA electroporation was applied for nonviral transfection of different types of human DCs, including monocyte-derived DCs (Mo-DCs), CD34(+) progenitor-derived DCs (34-DCs) and Langerhans cells (34 LCs). High-level transgene expression by mRNA electroporation was obtained in more than 50% of all DC types. mRNA-electroporated DCs retained their phenotype and maturational potential. Importantly, DCs electroporated with mRNA-encoding Melan-A strongly activated a Melan-A-specific cytotoxic T lymphocyte (CTL) clone in an HLA-restricted manner and were superior to mRNA-lipofected or -pulsed DCs. Optimal stimulation of the CTL occurred when Mo-DCs underwent maturation following mRNA transfection. Strikingly, a nonspecific stimulation of CTL was observed when DCs were transfected with plasmid DNA. The data clearly demonstrate that Mo-DCs electroporated with mRNA efficiently present functional antigenic peptides to cytotoxic T cells. Therefore, electroporation of mRNA-encoding tumor antigens is a powerful technique to charge human dendritic cells with tumor antigens and could serve applications in future DC-based tumor vaccines. PMID- 11418463 TI - Genetically modified bone marrow continuously supplies anti-inflammatory cells and suppresses renal injury in mouse Goodpasture syndrome. AB - In chronic inflammation, macrophages and neutrophils, which are derived from bone marrow, play a pivotal role. Therefore, reconstitution of bone marrow with anti inflammatory stem cells may modify inflammation. In this study, transplantation based gene therapy was applied to glomerular inflammation for a long-lasting suppression of the glomerular damage seen in chronic nephritis. Bone marrow cells were harvested from male donor mice, which had received 5-fluorouracil 3 days previously, and transduced with an interleukin 1 (IL-1) receptor antagonist (IL 1Ra) or a mock gene using a retrovirus vector. After confirmation that transduced cells possessed the transgene at approximately 0.7 copies per cell and secreted recombinant IL-1Ra, these cells were infused into sublethally irradiated (6 Gy) female recipients once daily for 4 consecutive days. These female recipient mice had the male Y antigen in bone marrow, liver, and spleen, and 10% to 20% of their spleen cells possessed the transgene even 8 weeks after transplantation. Glomerulonephritis was then induced in these mice. Renal function and histology were retarded in the mice whose bone marrow was reconstituted with IL-1Ra producing cells compared with mock transduced cells. In situ hybridization using a Y painting probe revealed that transplanted donor cells were recruited into the glomerulus upon induction of nephritis, suggesting therapeutic effects were channeled through the secretion of IL-1Ra from these cells. Furthermore, the survival rate after a second challenge with nephrotoxic antibody was significantly improved in the IL-1Ra chimera. These results suggest that reconstitution of bone marrow for continuous supply of anti-inflammatory cells may be a useful strategy for the treatment of chronic inflammation. PMID- 11418464 TI - Lack of neighborhood effects from a transcriptionally active phosphoglycerate kinase-neo cassette located between the murine beta-major and beta-minor globin genes. AB - For the treatment of beta-globin gene defects, a homologous recombination mediated gene correction approach would provide advantages over random integration-based gene therapy strategies. However, "neighborhood effects" from retained selectable marker genes in the targeted locus are among the key issues that must be taken into consideration for any attempt to use this strategy for gene correction. An Ala-to-Ile mutation was created in the beta6 position of the mouse beta-major globin gene (beta(6I)) as a step toward the development of a murine model system that could serve as a platform for therapeutic gene correction studies. The marked beta-major gene can be tracked at the level of DNA, RNA, and protein, allowing investigation of the impact of a retained phosphoglycerate kinase (PGK)-neo cassette located between the mutant beta-major and beta-minor globin genes on expression of these 2 neighboring genes. Although the PGK-neo cassette was expressed at high levels in adult erythroid cells, the abundance of the beta(6I) mRNA was indistinguishable from that of the wild-type counterpart in bone marrow cells. Similarly, the output from the beta-minor globin gene was also normal. Therefore, in this specific location, the retained, transcriptionally active PGK-neo cassette does not disrupt the regulated expression of the adult beta-globin genes. (Blood. 2001;98:65-73) PMID- 11418465 TI - Rescue of the colony-stimulating factor 1 (CSF-1)-nullizygous mouse (Csf1(op)/Csf1(op)) phenotype with a CSF-1 transgene and identification of sites of local CSF-1 synthesis. AB - Colony-stimulating factor 1 (CSF-1) regulates the survival, proliferation, and differentiation of mononuclear phagocytes. It is expressed as a secreted glycoprotein or proteoglycan found in the circulation or as a biologically active cell-surface glycoprotein. To investigate tissue CSF-1 regulation, CSF-1-null Csf1(op)/Csf1(op) mice expressing transgenes encoding the full-length membrane spanning CSF-1 precursor driven by 3.13 kilobases of the mouse CSF-1 promoter and first intron were characterized. Transgene expression corrected the gross osteopetrotic, neurologic, weight, tooth, and reproductive defects of Csf1(op)/Csf1(op) mice. Detailed analysis of one transgenic line revealed that circulating CSF-1, tissue macrophage numbers, hematopoietic tissue cellularity, and hematopoietic parameters were normalized. Tissue CSF-1 levels were normal except for elevations in 4 secretory tissues. Skin fibroblasts from the transgenic mice secreted normal amounts of CSF-1 but also expressed some cell surface CSF-1. Also, lacZ driven by the same promoter/first intron revealed beta galactosidase expression in hematopoietic, reproductive, and other tissue locations proximal to CSF-1 cellular targets, consistent with local regulation by CSF-1 at these sites. These studies indicate that the 3.13-kilobase promoter/first intron confers essentially normal CSF-1 expression. They also pinpoint new cellular sites of CSF-1 expression, including ovarian granulosa cells, mammary ductal epithelium, testicular Leydig cells, serous acinar cells of salivary gland, Paneth cells of the small intestine, as well as local sites in several other tissues. PMID- 11418466 TI - Platelet characteristics in patients with X-linked macrothrombocytopenia because of a novel GATA1 mutation. AB - A new mutation is described in the X-linked gene GATA1, resulting in macrothrombocytopenia and mild dyserythropoietic features but no marked anemia in a 4-generation family. The molecular basis for the observed phenotype is a substitution of glycine for aspartate in the strictly conserved codon 218 (D218G) of the amino-terminal zinc finger loop of the transcription factor GATA1. Zinc finger interaction studies demonstrated that this mutation results in a weak loss of affinity of GATA1 for its essential cofactor FOG1, whereas direct D218G-GATA1 binding to DNA was normal. The phenotypic effects of this mutation in the patients' platelets have been studied. Semiquantitative RNA analysis, normalized for beta-actin messenger RNA, showed extremely low transcription of the GATA1 target genes GPIbbeta and GPIX but also a significantly lower expression of the nondirectly GATA1-regulated Gsalpha gene, suggestive of incomplete megakaryocyte maturation. In contrast, GPIIIa expression was close to normal in agreement with its early appearance during megakaryocyte differentiation. Flow cytometric analysis of patient platelets confirmed the existence of a platelet population with abnormal size distribution and reduced GPIb complex levels but with normal GPIIIa expression. It also showed the presence of very immature platelets lacking almost all membrane glycoproteins studied (GPIbalpha, GPIbbeta, GPIIIa, GPIX, and GPV). Patients' platelets showed weak ristocetin-induced agglutination, compatible with the disturbed GPIb complex. Accordingly, electron microscopy of the patients' platelets revealed giant platelets with cytoplasmic clusters consisting of smooth endoplasmic reticulum and abnormal membrane complexes. In conclusion, GATA1 mutations can lead to isolated X-linked macrothrombocytopenia without anemia. PMID- 11418467 TI - Highly abundant genes in the transcriptosome of human and baboon CD34 antigen positive bone marrow cells. AB - Nonhuman primates are useful large animal model systems for the in vivo study of hematopoietic stem cell biology. To better understand the degree of similarity of the hematopoietic systems between humans and baboons, and to explore the relevance of such studies in nonhuman primates to humans, this study was designed to compare the global gene expression profile of bone marrow CD34(+) cells isolated from these 2 species. Human complementary DNA (cDNA) filter arrays containing 25 920 human cDNAs were surveyed for this purpose. The expression pattern and relative gene abundance of the 2 RNA sources were similar, with a correlation coefficient of 0.87. A total of 15 970 of these cDNAs were expressed in human CD34(+) cells, of which the majority (96%) varied less than 3-fold in their relative level of expression between human and baboon. Reverse transcriptase-polymerase chain reaction analysis of selected genes confirmed that expression was comparable between the 2 species. No species-restricted transcripts have been identified, further reinforcing the high degree of similarity between the 2 populations. A subset of 1554 cDNAs, which are expressed at levels 100-fold and greater than background, is described, which includes 959 expressed sequence tags and uncharacterized cDNAs, and 595 named genes, including many that are clearly involved in hematopoiesis. The cDNAs reported here represent a selection of some of the most highly abundant genes in hematopoietic cells and provide a starting point to develop a profile of the transcriptosome of CD34(+) cells. PMID- 11418468 TI - Novel structurally altered P(2X1) receptor is preferentially activated by adenosine diphosphate in platelets and megakaryocytic cells. AB - Experimental and clinical data suggest the presence of multiple types of adenosine diphosphate (ADP) receptors, one coupled to ligand-gated cation channels (P(2X)) and others coupled to G-protein-coupled (P(2Y)) receptors. This report identifies cDNA for a structurally altered P(2X1)-like receptor in megakaryocytic cell lines (Dami and CMK 11-5) and platelets that, when transfected into nonresponsive 1321 cells, confers a specific sensitivity to ADP with the pharmacologic rank order of ADP > > ATP > > > alpha,beta-methylene-ATP as measured by Ca(++) influx. This receptor (P(2X1del)) contains a deletion of 17 amino acids (PALLREAENFTLFIKNS) that includes an NFT consensus sequence for N linked glycosylation. Glycosylated forms of the P(2X1del) and P(2X1wt) receptors were indistinguishable electrophoretically by Western blot or by immunoprecipitation using available antihuman and antirat antibodies. These results indicate that the expression of the P(2X1del) receptor results in an influx of Ca(++) induced by ADP. Expression of P(2X1del) receptor homomeric subunits is sufficient to express a receptor preferentially activated by ADP and suggests that this altered form, alone or in combination with P(2X1wt) receptors, is a component of an ADP-activated ion channel. PMID- 11418469 TI - Platelet/polymorphonuclear leukocyte adhesion: a new role for SRC kinases in Mac 1 adhesive function triggered by P-selectin. AB - Adhesion of polymorphonuclear leukocytes (PMNLs) to activated platelets requires a P-selectin-triggered, tyrosine kinase-dependent adhesiveness of Mac-1 and is accompanied by tyrosine phosphorylation of a 110-kd protein (P-110) in PMNLs. Inhibitors of SRC tyrosine kinases were found to inhibit PMNL adhesion to activated platelets or to P-selectin expressing Chinese hamster ovary (CHO-P) cells and the tyrosine phosphorylation of P-110. Adhesion of PMNLs to activated platelets or to CHO-P cells stimulated activity of LYN and HCK. Monoclonal antibody blockade of P-selectin or beta2-integrins reduced the activation of both kinases. In PMNLs either adherent to platelets or aggregated by P-selectin-IgG chimera, Mac-1 was rapidly redistributed to the Triton X-100-insoluble cytoskeletal fraction, and large clusters of Mac-1 colocalized with patches of F actin at the sites of cell-cell contact. In PMNLs stimulated by P-selectin-IgG chimera, SRC kinase inhibition impaired Mac-1 clustering, F-actin accumulation, and CD18 redistribution to the cytoskeleton. Disruption of the actin filament network by cytochalasin D prevented PMNL-platelet adhesion and P-selectin-induced PMNL aggregation and impaired the clustering of Mac-1. In agreement with the requirement for the beta2-integrin in the functional up-regulation of LYN and HCK, integrin blockade by monoclonal antibodies resulted in a complete inhibition of P-selectin-induced Mac-1 clustering and F-actin accumulation. Taken together, the results indicate that, after an initial P-selectin-triggered beta2-integrin interaction with the ligand, SRC kinases are activated and allow the remodeling of cytoskeleton-integrin linkages and integrin clustering that finally strengthen cell-cell adhesion. This model highlights a new role for SRC kinases in a regulatory loop by which the Mac-1 promotes its own adhesive function. PMID- 11418470 TI - Assembly of a fibronectin matrix by adherent platelets stimulated by lysophosphatidic acid and other agonists. AB - Lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P) are agonists of the endothelial differentiation gene (Edg) family of G-protein-coupled receptors. LPA and S1P are generated by platelet activation during blood coagulation. Both lipids induce assembly of exogenous fibronectin (FN) by fibroblasts. This study examined whether LPA and S1P stimulate binding and assembly of fluoresceinated FN (FITC-FN) by adherent platelets. LPA enhanced deposition of FITC-FN into linear arrays overlying platelet surfaces and on edges of platelets adherent to FN or vitronectin (VN). Deposition was greater when platelets were adherent to FN than to VN and was elicited by platelet agonists with the following order of potency: thrombin > LPA = ADP (adenosine diphosphate) > S1P. The linear pattern of FITC-FN deposition was different from the more diffuse pattern of Alexa-fibrinogen (Alexa FGN) binding to adherent platelets. FITC-FN was deposited by adherent platelets that had dense arrays of cytoskeletal actin when stained with rhodamine phalloidin. The 70-kd N-terminal fragment of FN or L8 monoclonal antibody to a self-association domain of FN abolished deposition of FITC-FN but had no effect on binding of Alexa-FGN. Conversely, integrilin did not attenuate deposition of FITC-FN but abolished binding of Alexa-FGN. RGDS (Arg-Gly-Asp-Ser) or antibodies to alpha5beta1 or alphaIIbbeta3 integrins caused a partial decrease in LPA induced deposition of FITC-FN. Correlative electron microscopy with anti-FITC coupled to gold beads revealed linear arrays on platelet surfaces associated with less than 20-nm-diameter filaments. These observations demonstrate that LPA, thrombin, ADP, and S1P induce adherent platelets to bind and assemble FN and suggest that platelets may contribute to early deposition of FN matrix after vascular injury. PMID- 11418471 TI - Defective binding of factor XI-N248 to activated human platelets. AB - Variants of factor XI containing Gln226 to Arg (Q226 to R) and Ser248 to Asn (S248 to N) substitutions were first identified in an African American family with a history of excessive bleeding. The substitutions have recently been identified in unrelated individuals, suggesting they are relatively common. Both amino acids are located in the third apple domain of factor XI, an area implicated in binding interactions with factor IX and activated platelets. Recombinant factor XI-R226 and factor XI-N248 were compared with wild-type factor XI in assays for factor IX activation or platelet binding. Factor XI-R226 activates factor IX with a Michaelis-Menten constant (K(m)) about 5-fold greater than wild-type protein. The catalytic efficiency of factor IX activation is similar to wild-type protein, however, due to an increase in the turnover number (k(cat)) for the reaction. Iodinated factor XI-N248 binds to activated platelets with a dissociation constant (K(d)) more than 5-fold higher than wild-type protein (55 nM and 10 nM, respectively). Activation of factor XI-N248 by thrombin in the presence of activated platelets is slower and does not progress to the same extent as activation of the wild-type protein under similar conditions. Factor XI-N248 activates factor IX normally in a purified protein system and has relatively normal activity in activated partial thromboplastin time (aPTT) assays. Factor XI-N248 is the first factor XI variant described with a clear functional difference compared with wild-type protein. Importantly, the defect in platelet binding would not be detected by routine clinical evaluation with an aPTT assay. PMID- 11418472 TI - Immunodominant epitopes on glycoprotein IIb-IIIa recognized by autoreactive T cells in patients with immune thrombocytopenic purpura. AB - It was recently reported that autoreactive CD4(+) T cells to glycoprotein IIb IIIa (GPIIb-IIIa) mediate antiplatelet autoantibody production in patients with immune thrombocytopenic purpura (ITP). To further examine the antigenic specificity of the GPIIb-IIIa-reactive T cells, 6 recombinant fragments encoding different portions of GPIIbalpha or GPIIIa were generated and tested for their ability to stimulate antigen-specific T-cell proliferation and anti-GPIIb-IIIa antibody production in vitro. T cells from the peripheral blood of 25 patients with ITP and 10 healthy donors proliferated in response to recombinant GPIIb-IIIa fragments in various combinations. The amino-terminal portions of both GPIIbalpha and GPIIIa (IIbalpha18-259 and IIIa22-262) were frequently recognized (60% and 64%, respectively) compared with other fragments (4%-28%) in patients with ITP, but this tendency was not detected in healthy donors. In subsequent analyses in patients with ITP, T-cell reactivities to IIbalpha18-259 and IIIa22-262 were consistently detected, whereas those to other fragments were sometimes lost. In vitro antigenic stimulation of peripheral blood mononuclear cells with IIbalpha18 259 or IIIa22-262 promoted the synthesis of anti-GPIIb-IIIa antibodies in patients with ITP, but not in healthy donors. Of 15 CD4(+) T-cell lines specific for platelet-derived GPIIb-IIIa generated from 5 patients with ITP, 13 lines recognized IIbalpha18-259, IIIa22-262, or both. T-cell lines reactive to IIbalpha18-259 or IIIa22-262 promoted the production of anti-GPIIb-IIIa antibodies that were capable of binding to normal platelet surfaces. These results indicate that the immunodominant epitopes recognized by pathogenic CD4(+) T cells in patients with ITP are located within the amino-terminal portions of both GPIIbalpha and GPIIIa. PMID- 11418473 TI - Surgical and physical stress increases circulating blood dendritic cell counts independently of monocyte counts. AB - Dendritic cells (DCs) are specialized antigen-presenting cells that have the unique ability to initiate a primary immune response. The effect of physiologic stress on circulating blood DCs has thus far not been studied. In this study, we applied a recently developed method of counting blood DCs to test the hypothesis that significant stress to the body such as surgery and exercise might induce measurable changes in the DC numbers, subsets, phenotype, and function. Twenty six patients scheduled for elective laparoscopic cholecystectomy, 4 for elective hysterectomy, 56 controls, and 5 volunteers who underwent a stress exercise test were enrolled in the study. Absolute DC counts increased acutely (71.7% +/- 11% [SEM], P =.0001) in response to the stress of surgery and dropped below preoperative levels (-25% +/- 14% [SEM], P =.05) on days 2-3. The perioperative DC subset balance remained constant. Interestingly, DC counts changed independently of monocyte counts. Exercise also induced a rise in DC counts but coincidentally with monocyte counts. Surprisingly, no phenotypic or functional activation of DCs was seen in either stress situations in vivo. DCs are rapidly mobilized into the circulation in response to surgical and exercise stress, which may serve to prepare the host's immune defenses against trauma. The independent regulation of the DC and monocyte counts reinforces the distinction between these 2 cell populations. PMID- 11418474 TI - Dysfunctional Epstein-Barr virus (EBV)-specific CD8(+) T lymphocytes and increased EBV load in HIV-1 infected individuals progressing to AIDS-related non Hodgkin lymphoma. AB - Acquired immunodeficiency syndrome-related non-Hodgkin lymphomas (AIDS-NHL) are thought to arise because of loss of Epstein-Barr Virus (EBV)-specific cellular immunity. Here, an investigation was done to determine whether cellular immunity to EBV is lost because of physical loss or dysfunction of EBV-specific cytotoxic T cells. Data on EBV-specific cellular immunity were correlated with EBV load. For comparison, individuals who progressed to AIDS with opportunistic infections (AIDS-OI) and long-term asymptomatics (LTAs) were studied. The number of virus specific T cells was detected using tetrameric HLA-EBV-peptide complexes; function of these EBV-specific T cells was determined using the interferon-gamma (IFN-gamma) Elispot assay. It was observed that EBV-specific CD8(+) T cells were present in normal numbers in human immunodeficiency virus (HIV)-infected individuals. However, their functional capacity was decreased compared with HIV( ) individuals. In AIDS-NHL patients, EBV-specific T cells were not physically lost in the course of HIV-1 infection but showed progressive loss of their capability to produce IFN-gamma in response to EBV peptides. This loss of function correlated with lower CD4(+) T-cell numbers and was accompanied by increasing EBV load. In HIV-1-infected LTA individuals, in whom CD4(+) T-cell numbers were maintained, and progressors to AIDS-OI, IFN-gamma-producing EBV specific T cells were stable and EBV load remained stable or decreased in the course of HIV infection, suggestive of immune control. Our data indicate that functional loss of EBV-specific CD8(+) T cells with a concomitant increase in EBV load may play a role in the pathogenesis of AIDS-NHL. PMID- 11418475 TI - CD8 T cells specific for human immunodeficiency virus, Epstein-Barr virus, and cytomegalovirus lack molecules for homing to lymphoid sites of infection. AB - CD8 T cells are classified as naive, effector, or memory cells on the basis of CD45RA, CD62L, and CCR7 expression. Sequential engagement of cell-surface CD62L and CCR7 receptors is required for efficient trafficking to lymphoid tissue by means of high endothelial venules. Naive CD8 T cells are CCR7(+)CD62L(+) CD45RA(+), whereas long-term memory cells are CCR7(+)CD62L(+)CD45RA(-). Effector cytotoxic T cells are thought to be CCR7(-)CD45RA(+). The distribution of CD8 subsets and cytolytic protein expression in healthy donors and donors seropositive for human immunodeficiency virus (HIV) were compared. In HIV infected subjects, CCR7(-) CD8 T cells expanded at the expense of naive and long term memory cells. In both healthy donors and HIV-infected donors, CCR7(+) CD8 T cells were uniformly negative for perforin. In all subsets, perforin and granzyme A were not coordinately expressed, with perforin expression being more tightly regulated. The properties of CD8 T cells specific for cytomegalovirus, Epstein Barr virus (EBV), and HIV were studied by staining with major histocompatibility complex peptide tetramers. Antigen-specific cells for chronic infections with these viruses were uniformly CCR7(-) and predominantly CD62L(-). In 2 HIV seropositive donors, 3- to 4-fold fewer EBV-tetramer-positive cells were present in lymph nodes compared with blood. Antigen-specific CD8 T cells are therefore preferentially excluded from lymphoid sites, even when infection is primarily in lymphoid tissue. This may protect lymphoid tissues from immunopathological changes but compromise immune defense against viruses, such as HIV and EBV, that target lymphocytes. HIV-specific CD8 T cells do not express CD45RA, whereas EBV- and CMV-specific CD8 T cells are heterogeneous in CD45RA(+) expression. Lack of CD45RA expression may indicate incomplete differentiation of HIV-specific CD8 T cells to cytotoxic T cells. PMID- 11418476 TI - Molecular and flow cytometric analysis of the Vbeta repertoire for clonality assessment in mature TCRalphabeta T-cell proliferations. AB - Clonality assessment through Southern blot (SB) analysis of TCRB genes or polymerase chain reaction (PCR) analysis of TCRG genes is important for diagnosing suspect mature T-cell proliferations. Clonality assessment through reverse transcription (RT)-PCR analysis of Vbeta-Cbeta transcripts and flow cytometry with a Vbeta antibody panel covering more than 65% of Vbeta domains was validated using 28 SB-defined clonal T-cell receptor (TCR)alphabeta(+) T-ALL samples and T-cell lines. Next, the diagnostic applicability of the V(beta) RT PCR and flow cytometric clonality assays was studied in 47 mature T-cell proliferations. Clonal Vbeta-Cbeta RT-PCR products were detected in all 47 samples, whereas single Vbeta domain usage was found in 31 (66%) of 47 patients. The suspect leukemic cell populations in the other 16 patients showed a complete lack of Vbeta monoclonal antibody reactivity that was confirmed by molecular data showing the usage of Vbeta gene segments not covered by the applied Vbeta monoclonal antibodies. Nevertheless, this could be considered indirect evidence for the "clonal" character of these cells. Remarkably, RT-PCR revealed an oligoclonal pattern in addition to dominant Vbeta-Cbeta products and single Vbeta domain expression in many T-LGL proliferations, providing further evidence for the hypothesis raised earlier that T-LGL derive from polyclonal and oligoclonal proliferations of antigen-activated cytotoxic T cells. It is concluded that molecular Vbeta analysis serves to assess clonality in suspect T-cell proliferations. However, the faster and cheaper Vbeta antibody studies can be used as a powerful screening method for the detection of single Vbeta domain expression, followed by molecular studies in patients with more than 20% single Vbeta domain expression or large suspect T-cell populations (more than 50%-60%) without Vbeta reactivity. PMID- 11418477 TI - Rapamycin induces apoptosis in monocyte- and CD34-derived dendritic cells but not in monocytes and macrophages. AB - Rapamycin (Rapa), a recently introduced immunosuppressive drug, seems to be effective in preventing acute allograft rejection. Although its antiproliferative effect on T lymphocytes has been investigated extensively, its effect on the initiators of the immune response, the dendritic cells (DCs), is not known. Therefore, the effect of Rapa on monocyte- (mo-DCs) and CD34(+)-derived DCs in vitro but also on other myeloid cell types, including monocytes and macrophages, was examined. The present study shows that Rapa does not affect phenotypic differentiation and CD40L-induced maturation of mo-DCs. However, Rapa dramatically reduced cell recovery (40%-50%). Relatively low concentrations of Rapa (10(-9) M) induced apoptosis in both mo-DCs and CD34(+)-derived DCs, as visualized by phosphatidylserine exposure, nuclear condensation and fragmentation, and DNA degradation. In contrast, Rapa did not affect freshly isolated monocytes, macrophages, or myeloid cell lines. The sensitivity to Rapa induced apoptosis was acquired from day 2 onward of mo-DC differentiation. Rapa exerts its apoptotic effect via a reversible binding to the cytosolic receptor protein FKBP-12, as demonstrated in competition experiments with FK506, which is structurally related to Rapa. Partial inhibition of Rapa-induced apoptosis was obtained by addition of ZVAD-fmk, which implies caspase-dependent and caspase independent processes. The fact that Rapa exerts a specific effect on DCs but not on monocytes and macrophages might contribute to the unique actions of Rapa in the prevention of allograft rejection and other immune responses. PMID- 11418478 TI - CD38 expression as an important prognostic factor in B-cell chronic lymphocytic leukemia. AB - CD38 is a transmembrane glycoprotein expressed on the surface of leukemic cells in a significant percentage of patients with B-cell chronic lymphocytic leukemia (B-CLL). A recent study suggested that CD38 expression has prognostic value in CLL. Peripheral blood samples from 218 patients with B-CLL were analyzed by flow cytometry for CD38 expression on CD5/19(+) leukemic cells. Various patient characteristics were studied including age, sex, Rai and Binet stages, splenomegaly, hepatomegaly, hemoglobin (Hgb) level, beta-2 microglobulin (beta2M) level in the serum, number of nodal sites involved with disease, and length of survival. The Kaplan-Meier method was used to construct survival curves, and the log-rank statistic was used to compare these curves. CD38 was expressed in 20% or more of leukemic cells in 43% of the patients. Patients with high CD38 expression (20% or more) had significantly shorter survival times (P =.00005). Multivariate analyses showed that CD38 expression is an important prognostic factor associated with high incidence of lymph node involvement (P =.004), lower hemoglobin level (P =.001), hepatomegaly (P =.05), and high beta2M level (P =.00005). CD38 expression identified a group of patients with aggressive disease that was considered by Rai staging to be early-stage disease (Rai stages 0-II). Patients with CD38(+) samples have significantly aggressive disease regardless of their clinical stage. Measurement of CD38 expression by flow cytometry should become a routine test in the evaluation of patients with CLL. PMID- 11418479 TI - Endogenous CD28 expressed on myeloma cells up-regulates interleukin-8 production: implications for multiple myeloma progression. AB - CD28 is the major costimulatory molecule on T cells. CD28 activation, in conjunction with T-cell receptor engagement, up-regulates transcription of several cytokines, including interleukin-2 (IL-2), through transcriptional activation of the RE/AP composite element. Although CD28 is not normally expressed on B cells or plasma cells, more than 90% of extramedullary myelomas (a late stage B-cell neoplasm) express CD28. The functional significance of this is unknown. The results of this study demonstrate that CD28 stimulates transcriptional activation of RE/AP-based reporters in B cells and myeloma cells. However, CD28 stimulation does not up-regulate IL-2 production in myeloma cell lines, demonstrating that the IL-2 promoter may not be a relevant RE/AP containing target of CD28 in myelomas. Instead, an RE/AP composite element has been identified within the promoter of the IL-8 gene, a chemokine that promotes angiogenesis. Furthermore, stimulation of endogenous CD28 expressed by 3 myeloma cell lines increased IL-8 production. Therefore, the study demonstrates that CD28 is functional in myelomas to up-regulate transcription of endogenous genes, including IL-8. The proposal is made that aberrant expression of CD28 may play a role in the progression of multiple myeloma. PMID- 11418480 TI - The development of lymphomas in families with autoimmune lymphoproliferative syndrome with germline Fas mutations and defective lymphocyte apoptosis. AB - Lymphomas were studied in kindreds with autoimmune lymphoproliferative syndrome (ALPS; Canale-Smith syndrome), a disorder of lymphocyte homeostasis usually associated with germline Fas mutations. Fas (CD95/APO-1) is a cell surface receptor that initiates programmed cell death, or apoptosis, of activated lymphocytes. Lymphoma phenotype was determined by immunohistochemistry, frequency of CD3(+)CD4(-)CD8(-) T-cell-receptor alpha/beta cells by flow cytometry, nucleotide sequences of the gene encoding Fas (APT1, TNFRSF6), and the percentage of lymphocytes undergoing apoptosis in vitro. Of 223 members of 39 families, 130 individuals possessed heterozygous germline Fas mutations. Eleven B-cell and T cell lymphomas of diverse types developed in 10 individuals with mutations in 8 families, up to 48 years after lymphoproliferation was first documented. Their risk of non-Hodgkin and Hodgkin lymphomas, respectively, was 14 and 51 times greater than expected (each P <.001). Investigation of these 10 patients and their relatives with Fas mutations revealed that all had defective lymphocyte apoptosis and most had other features of ALPS. The tumor cells retained the heterozygous Fas mutations found in the peripheral blood and manifested defective Fas-mediated killing. These data implicate a role for Fas-mediated apoptosis in preventing B-cell and T-cell lymphomas. Inherited defects in receptor-mediated lymphocyte apoptosis represent a newly appreciated risk factor for lymphomas. PMID- 11418481 TI - EAF1, a novel ELL-associated factor that is delocalized by expression of the MLL ELL fusion protein. AB - The (11;19)(q23;p13.1) translocation in acute leukemia leads to the generation of a chimeric protein that fuses MLL to the transcriptional elongation factor ELL. A novel protein was isolated from a yeast 2-hybrid screen with ELL that was named EAF1 for ELL-associated factor 1. Using specific antibodies, the endogenous EAF1 and ELL proteins were coimmunoprecipitated from multiple cell lines. In addition, endogenous EAF1 also exhibited the capacity to interact with ELL2. Database comparisons with EAF1 identified a region with a high content of serine, aspartic acid, and glutamic acid residues that exhibited homology with the transcriptional activation domains of several translocation partner proteins of MLL, including AF4, LAF4, and AF5q31. A similar transcriptional activation domain has been identified in this region of EAF1. By confocal microscopy, endogenous EAF1 and ELL colocalized in a distinct nuclear speckled pattern. Transfection of the MLL ELL fusion gene delocalized EAF1 from its nuclear speckled distribution to a diffuse nucleoplasmic pattern. In leukemic cell lines derived from mice transplanted with MLL-ELL-transduced bone marrow, EAF1 speckles were not detected. Taken together, these data suggest that expression of the MLL-ELL fusion protein may have a dominant effect on the normal protein-protein interactions of ELL. PMID- 11418482 TI - Thalidomide and immunomodulatory derivatives augment natural killer cell cytotoxicity in multiple myeloma. AB - The antiangiogenic activity of thalidomide (Thal), coupled with an increase in bone marrow angiogenesis in multiple myeloma (MM), provided the rationale for the use of Thal in MM. Previously, the direct anti-MM activity of Thal and its analogues (immunomodulatory drugs, IMiDs) on MM cells was demonstrated, suggesting multiple mechanisms of action. In this study, the potential immunomodulatory effects of Thal/IMiDs in MM were examined. It was demonstrated that Thal/IMiDs do not induce T-cell proliferation alone but act as costimulators to trigger proliferation of anti-CD3-stimulated T cells from patients with MM, accompanied by an increase in interferon-gamma and IL-2 secretion. However, an increase in autologous T-cell killing of patient MM cells could not be demonstrated. A role for natural killer (NK)- and LAK-cell-mediated killing is suggested because IL-2-primed peripheral blood mononuclear cells (PBMCs) treated with Thal/IMiDs demonstrated significantly increased lysis of MM cell lines. Cold target inhibition assays suggested NK- rather than LAK-cell-mediated killing. Furthermore, this killing was not major histocompatibility complex-class restricted, and the depletion of CD56(+) cells blocked the drug-induced MM cell lysis. It was significant that increased killing of patient MM cells by autologous PBMCs treated with Thal/IMiDs was also observed. Although the in vivo relevance of NK-cell-mediated MM cell killing is unknown, phenotypic analysis performed in MM patients receiving Thal therapy demonstrated an increase in CD3( )CD56(+) cells in patients responding to therapy. Thus in vitro and in vivo data support the hypothesis that Thal may mediate its anti-MM effect, at least in part, by modulating NK cell number and function. PMID- 11418483 TI - Cyclin D3 at 6p21 is dysregulated by recurrent chromosomal translocations to immunoglobulin loci in multiple myeloma. AB - Reciprocal chromosomal translocations, which are mediated by errors in immunoglobulin heavy chain (IgH) switch recombination or somatic hypermutation as plasma cells are generated in germinal centers, are present in most multiple myeloma (MM) tumors. These translocations dysregulate an oncogene that is repositioned in proximity to a strong IgH enhancer. There is a promiscuous array of nonrandom chromosomal partners (and oncogenes), with the 3 most frequent partners (11q13 [cyclin D1]; 4p16 [FGFR3 and MMSET]; 16q23 [c-maf]) involved in nearly half of MM tumors. It is now shown that a novel t(6;14)(p21;q32) translocation is present in 1 of 30 MM cell lines and that this cell line uniquely overexpresses cyclin D3. The cloned breakpoint juxtaposes gamma 4 switch sequences with 6p21 sequences that are located about 65 kb centromeric to the cyclin D3 gene. By metaphase chromosome analysis, the t(6;14) (p21;q32) translocation was identified in 6 of 150 (4%) primary MM tumors. Overexpression of cyclin D3 messenger RNA (mRNA) was identified by microarray RNA expression analysis in 3 of 53 additional primary MM tumors, each of which was found to have a t(6;14) translocation breakpoint by interphase fluorescence in situ hybridization analysis. One tumor has a t(6;22)(p21;q11) translocation, so that cyclin D3 is bracketed by the IgL and IgH breakpoints. These results provide the first clear evidence for primary dysregulation of cyclin D3 during tumorigenesis. It is suggested that the initial oncogenic event for most MM tumors is a primary immunoglobulin translocation that dysregulates cyclin D1, cyclin D3, and other oncogenes to provide a proliferative stimulus to postgerminal center plasma cells. PMID- 11418484 TI - Intravenous injection of apoptotic leukocytes enhances bone marrow engraftment across major histocompatibility barriers. AB - Cross-tolerization of T lymphocytes after apoptotic cell uptake by dendritic cells may be involved in self-tolerance maintenance. Furthermore, immunosuppressive properties are attributed to apoptotic cells. This study evaluated the consequences of apoptotic leukocyte administration in a restrictive engraftment model of murine bone marrow (BM) transplantation. Sublethally irradiated recipients received a limited number of allogeneic BM, with or without irradiated apoptotic leukocytes of different origins. No graft-versus-host disease was observed. Whereas only a low proportion of mice receiving BM cells alone engrafted, addition of apoptotic irradiated leukocytes, independently of the origin (donor, recipient, third-party mice, as well as xenogeneic peripheral blood mononuclear cells), significantly enhanced engraftment. Similar results were obtained after infusion of leukocytes rendered apoptotic by UVB irradiation or by anti-Fas monoclonal antibody stimulation, thus confirming the role of apoptotic cells in engraftment facilitation. Overall, these results suggest that apoptotic leukocytes can nonspecifically facilitate allogeneic BM engraftment. Such a simple approach could be of interest in BM transplantation settings involving an important HLA donor/recipient disparity, a T-cell-depleted graft, or reduced conditioning regimen intensity. PMID- 11418485 TI - Pharmacogenetics of methotrexate: toxicity among marrow transplantation patients varies with the methylenetetrahydrofolate reductase C677T polymorphism. AB - This study investigated whether a polymorphism in the 5,10 methylenetetrahydrofolate reductase (MTHFR) gene (C677T) modifies responses to methotrexate (MTX) in patients undergoing bone marrow transplantation. About 10% to 12% of the population carry the MTHFR TT genotype (enzyme activity, 30% of wild type [CC]). Patients (n = 220) with chronic myelogenous leukemia underwent marrow allografts and were given a short course of MTX. MTX toxicity measures included the oral mucositis index (OMI), speed of engraftment (platelet and granulocyte counts), and bilirubin. Patients with lower MTHFR activity (TT genotype) had 36% higher mean OMI during days 1 to 18 (+5.7, P =.046) and 20% higher OMI between days 6 and 12 (+3.8, P =.27). Platelet counts recovered more slowly among patients with the TT genotype compared to wild type (24% slower recovery to 10 000 platelets/microL, P =.23; 34% slower to 20 000/microL, P =.08). Patients with decreased MTHFR activity appear at risk of higher MTX toxicity. Because of the high prevalence of the TT genotype, these results may have implications for MTX dosage. PMID- 11418486 TI - Fas ligand-induced caspase-1-dependent accumulation of interleukin-18 in mice with acute graft-versus-host disease. AB - Acute graft-versus-host disease (aGVHD), the fatal side effects of bone marrow transplantation, was shown to be accompanied by elevation of serum levels of interleukin 18 (IL-18). In this study, the mechanism underlying the accumulation of IL-18 in aGVHD in mice was investigated. Lethally irradiated recipients having transplantation with H-2 disparate donor splenocytes demonstrated aGVHD and contained markedly elevated serum levels of IL-18. In contrast, recipients having transplantation with gld/gld spleen cells, which lack functional Fas ligand (FasL), contained only normal ranges of IL-18, indicating FasL-mediated IL-18 release in aGVHD. The wild-type hosts engrafted with caspase-1-deficient cells revealed marked increases of IL-18 similar to those engrafted with wild-type cells, whereas caspase-1-deficient recipients engrafted with wild-type cells showed only a slight elevation of serum IL-18, indicating that IL-18 elevation is derived from host cells in a caspase-1-dependent manner. These results suggest FasL-mediated caspase-1-dependent IL-18 secretion in aGVHD in mice. PMID- 11418487 TI - Variable heavy-chain gene analysis of follicular lymphomas: subclone selection rather than clonal evolution over time. AB - To investigate B-cell receptor evolution in follicular lymphomas (FLs), immunoglobulin variable heavy chain (V(H)) gene regions of 3 FLs were analyzed at different time points. One FL with a high somatic mutation load and intraclonal V(H) gene diversity was investigated in situ. V(H) gene transcripts were amplified and sequenced from samples of approximately 50 tumor cells isolated from frozen tissue sections by laser microdissection. Interestingly, the mutation pattern of the prevalent subclone in the relapse biopsy was virtually identical to that of a subclone isolated by microdissection from the presentation biopsy 9 years earlier. In a second FL, proof was obtained that the subclone that dominated the relapse sample had already been present in the initial biopsy. The finding that subclones found in the relapses of these FLs had not evolved over time but were preexistent, challenges the concept of antigen-driven B-cell receptor evolution during disease course. PMID- 11418488 TI - Stable remission after administration of the receptor tyrosine kinase inhibitor SU5416 in a patient with refractory acute myeloid leukemia. AB - The small molecule receptor tyrosine kinase (RTK) inhibitor SU5416 targets the vascular endothelial growth factor receptor 2 and the stem cell factor receptor c kit. Herein is described the successful treatment of a 65-year-old woman with SU5416, in second relapse of acute myeloid leukemia (AML) and refractory toward standard chemotherapy regimens. After 12 weeks of treatment with SU5416, the blast cell counts (blood and bone marrow) decreased to undetectable levels and the peripheral blood cell counts normalized with the exception of the platelet count (50-80 x 10(9)/L [50-80 x 10(3)/microL]). The duration of the remission is longer than 4 months during maintenance therapy with SU5416. Microvessel density in the patient's bone marrow dropped from 33.4 to 12.3 microvessels/x500-field 8 weeks after SU5416 administration and remains in the normal range. This is the first report of a stable remission achieved after administration of the RTK inhibitor SU5416 in a patient with AML relapse. PMID- 11418489 TI - p16(INK4a) and p15(INK4b) gene methylations in plasma cells from monoclonal gammopathy of undetermined significance. AB - p15(INK4b) and p16(INK4a) proteins are cell cycle regulators involved in the inhibition of G1 phase progression. High frequency of methylation of both genes has been reported in multiple myeloma (MM), but it remains to be determined how and when these alterations contribute to tumorigenesis. Monoclonal gammopathy of undetermined significance (MGUS) represents an early disease stage in a fraction of MMs. Plasma cells from 33 patients with MGUS and 33 patients with MM were isolated and analyzed for p15(INK4b) and p16(INK4a) methylation by methylation specific polymerase chain reaction. Selective methylation was found in 19% for p16(INK4a), 36% for p15(INK4b), and 6.5% for both genes in MGUS, and frequencies were similar in MM suggesting that methylation of these genes is an early event, not associated with transition from MGUS to MM. p15(INK4b) and p16(INK4a) gene methylation might contribute to immortalization of plasma cells rather than malignant transformation in the natural history of MM. PMID- 11418490 TI - Female marmoset monkeys (Callithrix jacchus) can be identified from the chemical composition of their scent marks. AB - The present study analyzed 42 organic solvent extracts of scent mark pools from five dominant female common marmosets by gas chromatography (GC) and combined GC and mass spectrometry. We determined whether there were qualitative or quantitative differences between the chemical composition of scent marks from individual females. Gas chromatography and mass spectral analysis detected the same 162 chemicals in 86% (36/42) of scent mark pools from five dominant females. This near identical chemical composition of scent marks suggested there were few, if any, qualitative differences between the chemical composition of scent marks from individual females. Instead, quantitative differences in scent may provide the key factor distinguishing individual females. Using the relative concentration of highly volatile chemicals detected by GC in scent marks, linear discriminant analysis classified scent mark pools to their correct donor approximately 91% of the time. Such highly reliable statistical matching of scent to donor suggested that each individual female common marmoset has a unique ratio of highly volatile chemicals in their scent marks which may permit individual identification of females from odors in their scent alone. PMID- 11418491 TI - The influence of olfactory concept on the probability of detecting sub- and peri threshold components in a mixture of odorants. AB - The headspace of apple juice was analysed to obtain an ecologically relevant stimulus model mixture of apple volatiles. Two sets of volatiles were made up: a set of eight supra-threshold volatiles (MIX) and a set of three sub-threshold volatiles. These sets were used to test the hypothesis that sub-threshold components can change the quality of a familiar smelling mixture of odorants when added to this mixture. In order to test this hypothesis, three successive dilutions of the sub-threshold volatiles were prepared in such a way that the strongest was at the threshold concentration and the two lower concentrations were below the threshold. The detection probabilities of the sub-threshold components in a blank stimulus were compared with the detectabilities in MIX. The sub- and peri-threshold volatiles were detected no better in MIX than in a blank. On the contrary, sub- and peri-threshold volatiles were better detected alone than when added to MIX. However, when the group of subjects was split into two sub-groups, employing either a rough or a detailed concept definition of the target stimulus, respectively, the subjects with highly refined concepts were better able to detect the presence of sub-threshold volatiles in MIX than those with poorly refined stimulus concepts. The effect of stimulus concept definition occurred independently of the proportions of correct detections of sub-threshold volatiles in a blank. PMID- 11418492 TI - Hemispheric dominance of cortical activity evoked by focal electrogustatory stimuli. AB - Functional magnetic resonance imaging was used to observe cortical hemodynamic responses to electric taste stimuli applied separately to the right and left sides of the tongue tip. In 11 right-handed normal adults activation occurred primarily in the insular cortex, superior temporal lobe, inferior frontal lobe, including premotor regions, and in inferior parts of the postcentral gyrus. Unexpectedly, the location and laterality of activation were largely identical regardless of the side of the tongue stimulated. Activation in the superior insula, the presumed location of primary gustatory cortex, was predominantly, but not exclusively, in the right hemisphere, whereas central (more inferior) insular activations were more evenly bilateral. Right hemispheric dominance of activation also occurred in premotor regions (Brodmann areas 6 and 44), whereas left hemispheric dominance occurred only in the superior temporal cortex (Brodmann areas 22/42). The electric taste-evoked hemodynamic response pattern was more consistent with activation of the gustatory system than activation of somatosensory systems. The results suggest that the sites for cortical processing of electric taste information are dependent on hemispheric specialization. PMID- 11418493 TI - Genetic taste responses to 6-n-propylthiouracil among adults: a screening tool for epidemiological studies. AB - Genetically mediated taste responsiveness to 6-n-propylthiouracil (PROP) has been linked to reduced acceptance of some bitter foods. In this community-based study male (n = 364) and female (n = 378) adults enrolled in a self-help dietary intervention trial were screened for PROP taster status. Respondents, aged 18--70 years, were mailed filter papers impregnated with PROP or with aspartame solutions. They received instructions to rate taste intensity and hedonic preference using nine point category scales. Women rated PROP as more bitter than did men. Both sweetness and bitterness ratings were lower for older adults. Taste responsiveness to PROP was unrelated to body mass index in women or men. Higher bitterness ratings for PROP were weakly associated with higher sweetness ratings for aspartame, but were unrelated to sweet taste preferences. Successful administration of PROP filter papers by mail suggests new avenues for the screening of taste phenotypes in epidemiological studies. PMID- 11418494 TI - An animal model to assess aversion to intra-oral capsaicin: increased threshold in mice lacking substance p. AB - Despite the widespread consumption of products containing chemicals that irritate the oral mucosa, little is known about the underlying neural mechanisms nor is there a corresponding animal model of oral irritation. We have developed a rodent model to assess aversion to capsaicin in drinking water, using a paired preference paradigm. This method was used to test the hypothesis that the neuromodulator substance P (SP) plays a role in the detection of intra-oral capsaicin. 'Knockout' (KO) mice completely lacking SP and neurokinin A due to a disruption of the preprotachykinin A gene and a matched population of wild-type (WT) mice had free access to two drinking bottles, one containing water and the other capsaicin at various concentrations. Both KO and WT mice showed a concentration-dependent aversion to capsaicin. KO mice consumed significantly more capsaicin than WT at a single near threshold (1.65 microM) concentration, indicating that SP plays a limited role in the detection and rejection of oral irritants. PMID- 11418495 TI - Neural networks distinguish between taste qualities based on receptor cell population responses. AB - Response features of taste receptor cell action potentials were examined using an artificial neural network to determine whether they contain information about taste quality. Using the loose patch technique to record from hamster taste buds in vivo we recorded population responses of single fungiform papillae to NaCl (100 mM), sucrose (200 mM) and the synthetic sweetener NC-00274-01 (NC-01) (200 microM). Features of each response describing both burst and inter-burst characteristics were then presented to an artificial neural network for pairwise classification of taste stimuli. Responses to NaCl could be distinguished from those to both NC-01 and sucrose with accuracies of up to 86%. In contrast, pairwise comparisons between sucrose and NC-01 were not successful, scoring at chance (50%). Also, comparisons between two different concentrations of NaCl, 0.01 and 0.005 M, scored at chance. Pairwise comparisons using only those features that relate to the inter-burst behavior of the response (i.e. bursting rate) did not hinder the performance of the neural network as both sweeteners versus NaCl received scores of 75--85%. Comparisons using features corresponding to each individual burst scored poorly, receiving scores only slightly above chance. We then compared the sweeteners with varying concentrations of NaCl (0.1, 0.01, 0.005 and 0.001 M) using only those features corresponding to bursting rate within a 1 s time window. The neural network was capable of distinguishing between NaCl and NC-01 at all concentrations tested; while comparisons between NaCl and sucrose received high scores at all concentrations except 0.001 M. These results show that two different taste qualities can be distinguished from each other based solely on the bursting rates of action potentials in single taste buds and that this distinction is independent of stimulation intensity down to 0.001 M NaCl. These data suggest that action potentials in taste receptor cells may play a role in taste quality coding. PMID- 11418496 TI - An artificial sweetener stimulates the sweet taste in insect: dual effects of glycyrrhizin in Phormia regina. AB - Glycyrrhizin, found in the root of licorice (Glycyrrhizia glabra), has been used extensively as a non-sugar sweetener for humans and also as a medicine. As far as we know, the present work is the first report describing that a non-sugar sweetener for humans induces a sweet taste in insects. In behavioural experiments, we found that glycyrrhizin induced the feeding response, including full proboscis extension in the blowfly, Phormia regina. Glycyrrhizin also induced impulses of the sugar receptor cell in the labellar chemosensillum, which is highly specialized for the tastes of sugars and nucleotides. The optimum concentration of glycyrrhizin was 3.0 mM, which is much lower than that of sucrose. It has been established that multiple receptor sites, the pyranose receptor site (P site) and the furanose receptor site (F site), are present in the sugar receptor cell of the blowfly and the fleshfly. The inhibitors specific to the P site, starch and PCMB (p-chloromercuribenzoate), partially inhibited glycyrrhizin-induced responses but not levan (an inhibitor to the F site), indicating that the P site on the sugar receptor cell is involved in the glycyrrhizin action but not the F site. When 30 s stimulation with 3.0 mM glycyrrhizin was repeated with an interval of 3--10 min, the impulse frequency to the second stimulus was higher than that to the first one and doubled within 6 min. The first stimulus lasting longer than 10 s potentiated the impulse generation and reduced the adaptation rate during the second stimulus. These results suggest that, in addition to the action via the P site, an additional mechanism, possibly in the signal transduction cascade of the sugar receptor cell, may be involved in the action of glycyrrhizin. PMID- 11418497 TI - Vomeronasal epithelial cells of human fetuses contain immunoreactivity for G proteins, Go(alpha) and Gi(alpha 2). AB - Two G protein subfamilies, Go(alpha) and Gi(alpha 2), were identified and localized immunohistochemically in the vomeronasal organ (VNO) of 5-month-old human fetuses. Immunoreactivity for Go(alpha) and Gi(alpha 2) was present in a subset of vomeronasal epithelial cells. Prominent immunoreactivity was observed in apical processes and their apical terminals facing onto the vomeronasal lumen. Nerve fibers associated with the VNO exhibited intense immunoreactivity for Go(alpha) and weak immunoreactivity for Gi(alpha 2). Since Go(alpha) and Gi(alpha 2) are characteristically expressed and coupled with putative pheromone receptors in rodent vomeronasal receptor neurons, the present results suggest the possibility that vomeronasal epithelial cells containing Go(alpha) and Gi(alpha 2) in human fetuses are chemosensory neurons. PMID- 11418498 TI - The effect of lactic acid on odour-related host preference of yellow fever mosquitoes. AB - In a behavioural study we have investigated the role of lactic acid for the host preferences of yellow fever mosquitoes (Aedes aegypti) by comparing the attractiveness of rubbings from the hands of different human individuals and extracts obtained from skin rubbings from different mammals (Bos primigenius f. taurus, Capra aegagrus f. hircus, Felis silvestris f. catus and Homo sapiens). Certain human individuals were consistently more attractive to mosquitoes than others. Addition of lactic acid markedly increased the degree of attractiveness of formerly less attractive human odour samples and they were preferred over those which were originally the most attractive. There was almost no response to animal odour samples. In contrast to human samples, which contain a high amount of lactate, this compound could not be detected in samples from animals. When skin emanations from animals were combined with lactic acid, however, as many mosquitoes responded to odour samples of B. primigenius f. taurus and C. aegagrus f. hircus as did to human odours. All these data demonstrate that olfactory-based host preference of the anthropophilic mosquito A. aegypti is to a large extent due to differences in the amount of lactic acid in the odour samples. PMID- 11418499 TI - Detection and removal of an artefact fatty acid from the binding site of recombinant Bombyx mori pheromone-binding protein. AB - Recombinant Bombyx mori pheromone-binding protein (PBP), purified from an Escherichia coli expression system, has been found to contain (11Z)-octadecenoic acid (cis-vaccenic acid) as an artefact ligand. An efficient delipidation procedure is described to overcome what would appear to be a general problem with recombinant lepidopteran PBPs. PMID- 11418500 TI - Evaluation of the key odorants of foods by dilution experiments, aroma models and omission. AB - The state of the art in aroma analysis is reviewed with emphasis on aroma recombination studies using synthetic blends of odorants (aroma models) which have been prepared on the basis of analytical data. The model that matches the original aroma is the starting material for omission experiments which are performed to establish the odorants that actually contribute to the aroma. These experiments are discussed in detail for the aromas of two wine varieties, three olive oils of different provenance, French fries, boiled beef and coffee. The results indicate that odorants with higher odour activity values (OAV, the ratio of the concentration to the odour threshold) are frequently essential for the aroma. However, there are exceptions where odorants with high OAVs are suppressed in the aroma and compounds with lower OAVs are important contributors. These findings are discussed in the light of model experiments which have been reported in the literature to obtain an insight into the perceptual interactions of odorants in mixtures. PMID- 11418502 TI - Parallel-distributed processing in olfactory cortex: new insights from morphological and physiological analysis of neuronal circuitry. AB - A working hypothesis is proposed for piriform cortex (PC) and other olfactory cortical areas that redefines the traditional functional roles as follows: the olfactory bulb serves as the primary olfactory cortex by virtue of encoding 'molecular features' (structural components common to many odorant molecules) as a patchy mosaic reminiscent of the representation of simple features in primary visual cortex. The anterior olfactory cortex (that has been inappropriately termed the anterior olfactory nucleus) detects and stores correlations between olfactory features, creating representations (gestalts) for particular odorants and odorant mixtures. This function places anterior olfactory cortex at the level of secondary visual cortex. PC carries out functions that have traditionally defined association cortex--it detects and learns correlations between olfactory gestalts formed in anterior olfactory cortex and a large repertoire of behavioral, cognitive and contextual information to which it has access through reciprocal connections with prefrontal, entorhinal, perirhinal and amygdaloid areas. Using principles derived from artificial networks with biologically plausible parallel-distributed architectures and Hebbian synaptic plasticity (i.e. adjustments in synaptic strength based on locally convergent activity), functional proposals are made for PC and related cortical areas. Architectural features incorporated include extensive recurrent connectivity in anterior PC, predominantly feedforward connectivity in posterior PC and backprojections that connect distal to proximal structures in the cascade of olfactory cortical areas. Capabilities of the 'reciprocal feedforward correlation' architecture that characterizes PC and adjoining higher-order areas are discussed in some detail. The working hypothesis is preceded by a review of relevant anatomy and physiology, and a non-quantitative account of parallel-distributed principles. To increase the accessibility of findings for PC and to advertise its substantial potential as a model for experimental and modeling analysis of associative processes, parallels are described between PC and the hippocampal formation, inferotemporal visual cortex and prefrontal cortex. PMID- 11418503 TI - Receptive fields in the rat piriform cortex. AB - Current models of odor discrimination in mammals involve molecular feature detection by a large family of diverse olfactory receptors, refinement of molecular feature extraction through precise projections of olfactory receptor neurons to the olfactory bulb to form an odor-specific spatial map of molecular features across glomerular layer, and synthesis of these features into odor objects within the piriform cortex. This review describes our recent work on odor and spatial receptive fields within the anterior piriform cortex and compares these fields with receptive fields of their primary afferent, olfactory bulb mitral/tufted cells. The results suggest that receptive fields in the piriform cortex are ensemble in nature, highly dynamic, and may contribute to odor discrimination and odor memory. PMID- 11418504 TI - Neuromodulation and the functional dynamics of piriform cortex. AB - Acetylcholine and norepinephrine have a number of effects at the cellular level in the piriform cortex. Acetylcholine causes a depolarization of the membrane potential of pyramidal cells and interneurons, and suppresses the action potential frequency accommodation of pyramidal cells. Acetylcholine also has strong effects on synaptic transmission, suppressing both excitatory and inhibitory synaptic transmission. At the same time as it suppresses synaptic transmission, acetylcholine enhances synaptic modification, as demonstrated by experiments showing enhancement of long-term potentiation. Norepinephrine has similar effects. In this review, we discuss some of these different cellular effects and provide functional proposals for these individual effects in the context of the putative associative memory function of this structure. PMID- 11418505 TI - The rules of formation of the olfactory representations found in the orbitofrontal cortex olfactory areas in primates. AB - Approximately 35% of neurons in the orbitofrontal cortex taste and olfactory areas with olfactory responses provide a representation of odour that depends on the taste with which the odour has been associated previously. This representation is produced by a slowly acting learning mechanism that learns associations between odour and taste. Other neurons in the orbitofrontal cortex respond to both the odour and to the mouth feel of fat. The representation of odour thus moves for at least some neurons in the orbitofrontal cortex beyond the domain of physico-chemical properties of the odours to a domain where the ingestion-related significance of the odour determines the representation provided. Olfactory neurons in the primate orbitofrontal cortex decrease their responses to a food eaten to satiety, but remain responsive to other foods, thus contributing to a mechanism for olfactory sensory-specific satiety. It has been shown in neuroimaging studies that the human orbitofrontal cortex provides a representation of the pleasantness of odour, in that the activation produced by the odour of a food eaten to satiety decreases relative to another food-related odour not eaten in the meal. In the same general area there is a representation of the pleasantness of the smell, taste and texture of a whole food, in that activation in this area decreases to a food eaten to satiety, but not to a food that has not been eaten in the meal. PMID- 11418506 TI - Biochemical and genetic characterization of the action of triclosan on Staphylococcus aureus. AB - Triclosan, a widely used antibacterial agent, possesses potent activity against Staphylococcus aureus. This study reports on an investigation of the antibacterial target of triclosan in this pathogen. A strain of S. aureus overexpressing the enoyl-[acyl-carrier-protein] reductase (FabI), demonstrated by Western immunoblotting, gave rise to an increase in the MIC of triclosan, while susceptibilities to a range of unrelated antibacterials were unaffected. There are approximately 12 000 molecules of FabI per cell in mid-log phase growth. This number increased by approximately three- to four-fold in the S. aureus FabI overexpressor. Triclosan selectively inhibited the incorporation of [(14)C]acetate into TCA-precipitable product, an indicator of fatty acid biosynthesis. Furthermore, it inhibited de novo fatty acid biosynthesis in this organism. In vitro, triclosan inhibited recombinant, purified S. aureus FabI with an IC(50) of approximately 1 microM. The combination of these biochemical and genetic data provide further evidence that the mode of action of triclosan in S. aureus is via inhibition of FabI. PMID- 11418507 TI - Antimicrobial effects of positively charged surfaces on adhering Gram-positive and Gram-negative bacteria. AB - The infection of biomaterials is determined by an interplay of adhesion and surface growth of the infecting organisms. In this study, the antimicrobial effects on adhering bacteria of a positively charged poly(methacrylate) surface (xi potential +12 mV) were compared with those of negatively charged poly(methyl methacrylate) (-12 mV) and a highly negatively charged poly(methacrylate) (-18 mV) surface. Initial adhesion of Staphylococcus aureus ATCC 12600, Staphylococcus epidermidis HBH(2) 102, Escherichia coli O2K2 and Pseudomonas aeruginosa AK1 to these surfaces was measured in a parallel plate flow chamber in phosphate buffered saline. Adhering bacteria were allowed to multiply by perfusing the flow chamber with growth medium. All bacteria adhered most rapidly to the positively charged surface, but there was no subsequent surface growth of the Gram-negative strains. On the negatively charged surfaces, despite a slower initial adhesion, surface growth of the adhering bacteria was exponential for both Gram-positive and Gram-negative strains. These results suggest that positively charged biomaterial surfaces exert an antimicrobial effect on adhering Gram-negative bacteria, but not on Gram-positive ones. PMID- 11418508 TI - The effect of pharmacokinetics on the bactericidal activity of ciprofloxacin and sparfloxacin against Streptococcus pneumoniae and the emergence of resistance. AB - The pharmacokinetics of ciprofloxacin and sparfloxacin were simulated in vitro and the effects of pharmacodynamic parameters on bactericidal activity and the emergence of quinolone resistance were examined for Streptococcus pneumoniae. Simulated serum concentrations of ciprofloxacin 500 mg bd were more rapidly bactericidal than sparfloxacin 200 mg bd, despite lower values for the key pharmacodynamic parameters (AUC/MIC and C(max)/MIC). This was possibly related to the slower oral absorption of sparfloxacin, which delayed achievement of the MIC compared with ciprofloxacin. In addition, sparfloxacin was shown to have similar bactericidal activity to ciprofloxacin when tested at the same concentrations, despite its four-fold better potency in MIC terms. The emergence of resistance following exposure to ciprofloxacin appeared to be dependent on the C(max)/MIC ratio and the AUC above the MIC, but not the AUC/MIC ratio. Resistance (at least four-fold increase in MIC) developed when the C(max)/MIC ratio was less than four or the AUC above the MIC was less than 10, and the resulting cultures regrew fully. In contrast, pneumococci with a two- to four-fold increase in sparfloxacin MIC were selected in the presence of serum concentrations of sparfloxacin despite a C(max)/MIC ratio higher than 12, but these isolates remained clinically susceptible by breakpoint MIC and their growth was inhibited by repeated dosage of sparfloxacin. Nevertheless, the selection of pneumococci with reduced susceptibility, and the possibility of further mutation to highly resistant strains supports the use of quinolones that rapidly eradicate pneumococci at conventional doses and achieve concentrations, in both serum and tissues, which exceed at least 4 x MIC. PMID- 11418509 TI - Comparative in vitro antimicrobial activity of a new carbapenem, E1010, and tentative disc diffusion test interpretative criteria. AB - The susceptibility of 705 bacterial isolates representing 46 different species to E1010 (ER-35786), imipenem, meropenem and cefepime was determined by the NCCLS broth microdilution test. The MIC(90)s for E1010 were < or =1.0 mg/L for Enterobacteriaceae, fastidious Gram-negative bacteria, streptococci and anaerobes. E1010 was two- to four-fold more active than imipenem and meropenem against Pseudomonas aeruginosa, and four-fold more active than the other carbapenems against methicillin-resistant Staphylococcus aureus. Vancomycin resistant enterococci and most Enterococcus faecium were resistant to all four drugs tested. The NCCLS disc diffusion test was performed simultaneously on the non-fastidious organisms. Assuming the MIC breakpoints for E1010 will be the same as for the other carbapenems, the disc diffusion zone diameter breakpoints of imipenem and meropenem would also be applicable to E1010. PMID- 11418510 TI - Activity of non-fluorinated quinolones (NFQs) against quinolone-resistant Escherichia coli and Streptococcus pneumoniae. AB - The newly developed 8-methoxy, non-fluorinated quinolones (NFQs) were studied to elucidate their enzyme inhibitory activity against wild-type and mutant GyrA (Ser 83-->Trp) forms of Escherichia coli DNA gyrase. Using a DNA supercoiling inhibition assay, the NFQs were found to inhibit 50% (IC50) of the E. coli DNA gyrase activity in the 1.6-3.2 mg/L concentration range and were comparable to ciprofloxacin. However, against the GyrA (Ser-83-->Trp) mutant, the NFQs were approximately 16-fold more potent than ciprofloxacin. Antibacterial potency of the NFQs was investigated using clinical isolates of E. coli and penicillin resistant Streptococcus pneumoniae (PRSP), including strains with reduced susceptibility to quinolones. Against 20 uncharacterized clinical isolates of E. coli, the MIC90s of the NFQs were in the 0.125-0.25 mg/L range while those of ciprofloxacin, trovafloxacin, gatifloxacin and clinafloxacin were in the 0.016 0.125 mg/L range. Against clinical isolates with characterized mutations in gyrA and parC, PGE9262932, an NFQ, was two- to eight-fold more potent than ciprofloxacin. Against 23 clinical isolates of PRSP, the NFQs (MIC90 0.031-0.125 mg/L) were more potent than ciprofloxacin, trovafloxacin, and gatifloxacin (MIC90 0.25-2.0 mg/L), and at least as potent as clinafloxacin (MIC90 0.125 mg/L). Against S. pneumoniae strains with gyrA and parC mutations, the NFQs (MIC 0.125 1.0 mg/L) were more potent than ciprofloxacin, trovafloxacin and gatifloxacin (MIC 4-32 mg/L), and comparable to clinafloxacin (MIC 0.5-1 mg/L). PMID- 11418511 TI - Activity of levofloxacin and ciprofloxacin against urinary pathogens. AB - This study compares the antibacterial activities of levofloxacin and ciprofloxacin against recently isolated urinary tract pathogens, by evaluating their MICs and MBCs in accordance with NCCLS susceptibility tests, time-kill curves and interference with bacterial adhesion to uroepithelial cells. A total of 200 clinical isolates was tested, including the species Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Proteus vulgaris, Providencia rettgeri, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus and Staphylococcus epidermidis. All E. coli isolates were susceptible to levofloxacin and only one was resistant to ciprofloxacin, and there were no differences between beta-lactamase-positive and -negative strains. K. pneumoniae strains resistant to ciprofloxacin were also resistant to levofloxacin. Methicillin resistant S. aureus seemed to be less susceptible than methicillin-susceptible strains to these quinolones. S. epidermidis strains were susceptible to levofloxacin and ciprofloxacin, with the exception of two isolates. Incubation of S. aureus and E. coli with subinhibitory antimicrobial concentrations reduced their capacity to adhere to uroepithelial cells; this was statistically significant at 0.25 x MIC with respect to controls (P < 0.05). Inhibition of adhesion ranged from 36 to 43% when bacteria were incubated in the presence of 0.25 x MIC of levofloxacin and ciprofloxacin, and from 10 to 27% at 0.125 x MIC. These findings suggest that levofloxacin is an effective alternative to ciprofloxacin in the treatment of urinary tract infections and that sub inhibitory concentrations may contribute to efficacy. PMID- 11418512 TI - Antimicrobial susceptibility patterns of enterococci in intensive care units in Sweden evaluated by different MIC breakpoint systems. AB - Three hundred and twenty-two (322) clinical isolates were collected from patients admitted to intensive care units (ICUs) at eight Swedish hospitals between December 1996 and December 1998. Of the isolates, 244 (76%) were Enterococcus faecalis, 74 (23%) were Enterococcus faecium and four (1%) were other Enterococcus spp. MICs of ampicillin, imipenem, meropenem, piperacillin/tazobactam, ciprofloxacin, trovafloxacin, clinafloxacin, gentamicin, streptomycin, vancomycin, teicoplanin, quinupristin/dalfopristin, linezolid and evernimicin were determined by Etest. Susceptible and resistant isolates were defined according to the species-related MIC breakpoints of the British Society for Antimicrobial Chemotherapy (BSAC), the National Committee for Clinical Laboratory Standards (NCCLS) and the Swedish Reference Group for Antibiotics (SRGA). Tentative breakpoints were applied for new/experimental antibiotics. Multidrug resistance among enterococci in ICUs is not uncommon in Sweden, particularly among E. faecium, and includes ampicillin resistance and concomitant resistance to fluoroquinolones. Almost 20% of E. faecalis isolates showed high level resistance to gentamicin and concomitant resistance to fluoroquinolones. Vancomycin-resistant enterococci were only found sporadically. Among the new antimicrobial agents, linezolid and evernimicin showed the best activity against all enterococcal isolates. There was good concordance between the BSAC, NCCLS and SRGA breakpoints in detecting resistance. When applying the SRGA breakpoints for susceptibility, isolates were more frequently interpreted as intermediate. This might indicate earlier detection of emerging resistance using the SRGA breakpoint when the native population is considered susceptible, but with the risk that isolates belonging to the native susceptible population will be incorrectly interpreted as intermediate. PMID- 11418513 TI - Concentrations of gatifloxacin in plasma and pulmonary compartments following a single 400 mg oral dose in patients undergoing fibre-optic bronchoscopy. AB - The concentrations of gatifloxacin achieved after a single 400 mg oral dose were measured in plasma, epithelial lining fluid (ELF), alveolar macrophages (AMs) and bronchial mucosa (BM) using a microbiological assay. Fourteen patients undergoing fibre-optic bronchoscopy were studied. Mean plasma, ELF, AMs and BM concentrations, respectively, at 2, 4 and 12 h were as follows: 2 h: 3.96 mg/L, 6.00 mg/L, 69.10 mg/L, 6.24 mg/kg; 4 h: 3.22 mg/L, 6.16 mg/L, 77.32 mg/L, 5.32 mg/kg; 12 h: 1.74 mg/L, 2.98 mg/L, 61.95 mg/L, 3.00 mg/kg. These concentrations exceed the MIC(90)s for common respiratory pathogens such as Streptococcus pneumoniae (0.5 mg/L), Haemophilus influenzae (0.013 mg/L), Moraxella catarrhalis (0.05 mg/L), Chlamydia pneumoniae (0.125 mg/L) and Mycoplasma pneumoniae (0.06 mg/L). PMID- 11418514 TI - Resistance to itraconazole in Aspergillus nidulans and Aspergillus fumigatus is conferred by extra copies of the A. nidulans P-450 14alpha-demethylase gene, pdmA. AB - Triazoles selectively inhibit the cytochrome P-450-dependent C-14 lanosterol alpha-demethylase (P-450 14 alpha DM), a key enzyme in ergosterol biosynthesis in fungi. To investigate mechanisms of triazole resistance in a mould, we used Aspergillus nidulans, a genetically amenable model fungus closely related to more pathogenic members of the genus. We selected for genes that would give resistance to itraconazole following transformation with a high copy genomic library of A. nidulans. In all the resistant colonies that we isolated, resistance was conferred by extra copies of the A. nidulans P-450 14 alpha DM gene, pdmA. We determined that in A. nidulans, extra copies of pdmA increase the MIC for itraconazole 36 times over wild-type controls. Similarly, transformation of an Aspergillus fumigatus strain with pITZR1 resulted in increased resistance to itraconazole. Our results indicate that triazole resistance in clinical isolates of moulds may result from amplification or overexpression of the P-450 14 alpha DM and demonstrate the utility of A. nidulans as a promising model fungus for the analysis of drug resistance and susceptibility in the pathogenic fungus A. fumigatus. PMID- 11418515 TI - In vitro comparison of the antimycotic activity of a miconazole-HP-beta cyclodextrin solution with a miconazole surfactant solution. AB - The antimycotic activity of a new parenteral solution containing miconazole was compared with that of a marketed solution (Daktarin IV solution). This solution has been withdrawn from the Belgian market, probably because of toxic effects related to the presence of polyoxyl 35 castor oil. We propose a new formulation containing miconazole (10 mg/mL) (like the marketed solution), in combination with hydroxypropyl-beta-cyclodextrin and lactic acid. The MICs of these two solutions were determined by a broth microdilution method (based on NCCLS guidelines) for 67 yeasts and 50 filamentous fungi isolates. This study shows that the MICs obtained with these two solutions are not significantly different. PMID- 11418516 TI - Efficacy of aerosolized amphotericin B desoxycholate and liposomal amphotericin B in the treatment of invasive pulmonary aspergillosis in severely immunocompromised rats. AB - The effects of treatment with aerosolized amphotericin B desoxycholate and aerosolized liposomal amphotericin B were evaluated in severely immunosuppressed rats with invasive pulmonary aspergillosis. Aerosol treatment with amphotericin B desoxycholate consisted of a single dose (60 min) with amphotericin B concentrations in the nebulizer reservoir of 1, 2 and 4 mg/mL, respectively. For liposomal amphotericin B, aerosol treatment consisted of single, double or quadruple doses with a nebulizer reservoir concentration of 4 mg/mL of amphotericin B. Treatment, started at 30 h after inoculation, with aerosolized amphotericin B desoxycholate (nebulizer reservoir concentration 2 mg/mL) significantly prolonged survival of rats as compared with placebo-treated rats, whereas treatment with aerosolized amphotericin B desoxycholate with nebulizer reservoir concentration of 1 or 4 mg/mL did not have a significant effect on survival. Treatment with aerosolized liposomal amphotericin B significantly prolonged survival with all treatment regimens when compared with placebo-treated animals. Aerosol treatment did not prevent dissemination of the infection. The effects of amphotericin B desoxycholate and liposomal amphotericin B on pulmonary surfactant function were also evaluated in vitro. Amphotericin B desoxycholate inhibited surfactant function in a dose-dependent fashion. Liposomal amphotericin B had no detrimental effect on surface activity of surfactant. These results indicate that aerosol administration of amphotericin B, especially the liposomal formulation, could be an additional approach to optimizing treatment of invasive pulmonary aspergillosis. PMID- 11418517 TI - Itraconazole versus amphotericin B plus nystatin in the prophylaxis of fungal infections in neutropenic cancer patients. AB - The efficacy and safety of itraconazole oral solution and a combination of amphotericin B capsules plus nystatin oral suspension were compared in the prophylaxis of fungal infections in neutropenic patients. In an open, randomized, multicentre trial, 144 patients received itraconazole oral solution 100 mg bd, and 133 patients received amphotericin B 500 mg tds plus nystatin 2 MU qds. Overall, 65% of itraconazole-treated patients were considered to have had successful prophylaxis, compared with 53% in the polyene group. Proven deep fungal infections occurred in 5% of patients in each group. Fewer patients receiving itraconazole than amphotericin plus nystatin had superficial infections (3 versus 8%; P = 0.066). This trend in favour of itraconazole was seen in patients with profound neutropenia (neutrophil count <0.1 x 10(9) cells/L at least once) or prolonged neutropenia (neutrophil count <1.0 x 10(9) cells/L for >14 days). The median time to prophylactic failure was longer in the itraconazole group (37 days) than in the polyene group (34 days). The number of patients with fungal colonization (nose, sputum, stool) changed more favourably from baseline to endpoint in the itraconazole group than in the polyene group. Both treatments were safe and well tolerated; however, patients receiving amphotericin plus nystatin had a higher incidence of nausea and rash. In conclusion, itraconazole oral solution at doses of 100 mg bd and oral amphotericin B plus nystatin have similar prophylactic efficacy against fungal infections in neutropenic patients. On the basis of reduced incidence of superficial fungal infections, fungal colonization and specific adverse events, itraconazole may be the preferred treatment. PMID- 11418518 TI - In vitro effects of photodynamic therapy on Aspergillus fumigatus. AB - Photodynamic therapy is a technique for killing cells with visible light after pretreatment with a photosensitizing agent. We demonstrated significant in vitro fungicidal activity against Aspergillus fumigatus of the photosensitizer Green 2W, activated with 630 nm light. This effect was both inoculum- and light dose dependent. At a Green 2W concentration of 31.5 mg/L, there was complete killing of 2.7 x 10(1) cfu/mL with a light dose of 110 J/cm(2) and up to 2.7 x 10(6) cfu/mL with a light dose of 385 J/cm(2). PMID- 11418519 TI - The effect of amiloride on amphotericin B-induced hypokalaemia. AB - Amiloride was administered to 19 oncology patients exhibiting marked amphotericin B-induced electrolyte wasting. Mean serum potassium concentrations increased in the 5 days preceding and following administration (3.4 +/- 0.5 versus 3.9 +/- 0.8 mmol/L, P = 0.002). A trend towards decreased potassium supplementation was also observed (48.0 +/- 66.5 versus 29.4 +/- 43.2 mmol/day, P = 0.12). Amiloride is a therapeutic option to decrease potassium wasting in patients being treated with amphotericin B. PMID- 11418520 TI - Distribution of K103N and/or Y181C HIV-1 mutations by exposure to zidovudine and non-nucleoside reverse transcriptase inhibitors. AB - Our aim was to identify whether zidovudine has a role in the emergence of the K103N resistance mutation in the HIV-1 reverse transcriptase gene on non nucleoside reverse transcriptase inhibitors (NNRTIs). No difference was found in the exposure to zidovudine or major zidovudine mutations between the resistance patterns K103N-/Y181C+, K103N+/Y181C- and K103N+/ Y181C+, either in group A (patients on nevirapine and previously NNRTI naive) or in group B (on any NNRTI and experience of two or more NNRTIs including nevirapine). Group B patients had the highest prevalence of K103N+/Y181C+. In conclusion, zidovudine seems not to determine the emergence of K103N; however, there appears to be an accumulation of NNRTI resistance mutations with sequential use of NNRTIs. PMID- 11418521 TI - Elution characteristics of vancomycin, teicoplanin, gentamicin and clindamycin from calcium sulphate beads. AB - The in vitro release of vancomycin, teicoplanin, gentamicin and clindamycin from biodegradable calcium sulphate (CaSO(4)) carrier beads is described. All antibiotics showed prolonged release from the carrier beads, which was elevated during the first 24 h, with peak levels exceeding 2500 microg/bead. Doubling the antibiotic load of the beads revealed a more prolonged elution and a two-fold increase in antibiotic release. Local carrier-associated antibiotic treatment with CaSO(4) beads may prove to be effective in the management of chronic bone infections. PMID- 11418522 TI - Interpretative criteria and quality control parameters for telithromycin disc diffusion susceptibility tests. AB - In vitro studies were carried out with 4175 Gram-positive clinical isolates, comparing telithromycin broth microdilution MICs with disc diffusion zone diameters. Provisional interpretive criteria for testing staphylococci were > or =23 mm for susceptible and < or =19 mm for resistant. Because pneumococci and other streptococci require increased CO(2), the corresponding zone sizes are 3 mm smaller (> or =20 mm and < or =16 mm). Two collaborative studies were performed in order to propose the following quality control limits: 24-30 mm for Staphylococcus aureus ATCC 25923 and 27-33 mm for Streptococcus pneumoniae ATCC 49619. PMID- 11418523 TI - Susceptibility of the Bacteroides fragilis group to newer quinolones and other standard anti-anaerobic agents. AB - The susceptibilities of 200 clinical isolates of the Bacteroides fragilis group to four quinolones (moxifloxacin, clinafloxacin, trovafloxacin and ciprofloxacin) were determined, as well as to cefoxitin, clindamycin, metronidazole, imipenem and ticarcillin-clavulanic acid. The results for the latter five agents were compared with those of a study on 200 isolates done 6 years previously. Clinafloxacin and trovafloxacin were the most active agents tested with MIC90s lower than all other antimicrobials except imipenem. Susceptibility rates for imipenem, ticarcillin- clavulanic and metronidazole continue to be high, although resistant strains are emerging. For ticarcillin-clavulanic acid and metronidazole, MIC90s increased four- to eight-fold for the B. fragilis species between the two study periods. PMID- 11418524 TI - In vitro and in vivo efficacy of a novel fluoro-ketolide HMR 3562 against enterococci. AB - The in vitro activity of HMR 3562, a new 2-fluoro-ketolide drug, was investigated against 95 enterococci, including 36 vancomycin-resistant strains. HMR 3562 inhibited 90% of enterococci susceptible or resistant to erythromycin A at 0.005 and 0.6 mg/L, respectively. HMR 3562 was highly active in murine peritonitis induced by five enterococci, irrespective of resistance phenotype, displaying effective doses in the range 3.4-21.8 mg/kg. The results demonstrate the potential of HMR 3562 in the treatment of infections caused by multiresistant enterococci. PMID- 11418525 TI - The in vivo activity of olamufloxacin (HSR-903) in systemic and urinary tract infections in mice. AB - The in vivo activity of olamufloxacin (HSR-903), a new fluoroquinolone, was evaluated and compared with ciprofloxacin, sparfloxacin and levofloxacin. Olamufloxacin was active against systemic infection in mice inoculated with both Gram-positive and -negative bacteria. Olamufloxacin had equal efficacy for experimental urinary tract infections in mice caused by Pseudomonas aeruginosa. PMID- 11418526 TI - Dormancy and persistence in chronic infection: role of the general stress response in resistance to chemotherapy. PMID- 11418527 TI - Emergence of TetM-mediated tetracycline resistance in rural South Africa. PMID- 11418528 TI - Dominance of EMRSA-15 and -16 among MRSA causing nosocomial bacteraemia in the UK: analysis of isolates from the European Antimicrobial Resistance Surveillance System (EARSS). PMID- 11418529 TI - Extremely low prevalence of UK Staphylococcus aureus isolates with reduced susceptibility to vancomycin. PMID- 11418530 TI - Comparative in vitro study of the activity of moxifloxacin and other antibiotics against 150 strains of penicillin non-susceptible Streptococcus pneumoniae and against 110 strains of ampicillin-resistant Haemophilus influenzae isolated in 1999-2000 in Spain. PMID- 11418531 TI - In vitro activity of faropenem and 21 other compounds against 385 different genetically characterized isolates of antibiotic-resistant Streptococcus pneumoniae. PMID- 11418532 TI - Antibiotic resistance surveillance of Stenotrophomonas maltophilia, 1993-1999. PMID- 11418533 TI - Difficulties in the assay of cefamandole highlight the importance of specific methodologies in pharmacokinetic studies. PMID- 11418534 TI - Faecal concentrations of piperacillin and tazobactam in elderly patients. PMID- 11418536 TI - Reducing the frequency of errors in medicine using information technology. AB - BACKGROUND: Increasing data suggest that error in medicine is frequent and results in substantial harm. The recent Institute of Medicine report (LT Kohn, JM Corrigan, MS Donaldson, eds: To Err Is Human: Building a Safer Health System. Washington, DC: National Academy Press, 1999) described the magnitude of the problem, and the public interest in this issue, which was already large, has grown. GOAL: The goal of this white paper is to describe how the frequency and consequences of errors in medical care can be reduced (although in some instances they are potentiated) by the use of information technology in the provision of care, and to make general and specific recommendations regarding error reduction through the use of information technology. RESULTS: General recommendations are to implement clinical decision support judiciously; to consider consequent actions when designing systems; to test existing systems to ensure they actually catch errors that injure patients; to promote adoption of standards for data and systems; to develop systems that communicate with each other; to use systems in new ways; to measure and prevent adverse consequences; to make existing quality structures meaningful; and to improve regulation and remove disincentives for vendors to provide clinical decision support. Specific recommendations are to implement provider order entry systems, especially computerized prescribing; to implement bar-coding for medications, blood, devices, and patients; and to utilize modern electronic systems to communicate key pieces of asynchronous data such as markedly abnormal laboratory values. CONCLUSIONS: Appropriate increases in the use of information technology in health care- especially the introduction of clinical decision support and better linkages in and among systems, resulting in process simplification-could result in substantial improvement in patient safety. PMID- 11418537 TI - Consumer informatics supporting patients as co-producers of quality. AB - The track entitled "Consumer Informatics Supporting Patients as Co-Producers of Quality" at the AMIA Spring 2000 Congress was devoted to examining the new field of consumer health informatics. This area is developing rapidly, as worldwide changes are occurring in the organization and delivery of health care and in the traditional roles of patient and provider. This paper describes the key themes of the track; implications of the growing area of consumer health informatics; and recommendations for informatics research, design, and policy. Key themes that emerged from the panels and discussions involved changes in roles of consumers and providers; supporting a patient-provider-information technology partnership; virtual, not physical, structure for health care and health care information delivery; and health care as an integrated part of one's life. Panelists and participants at the Congress developed recommendations for informatics research, design, and policy, with an overarching focus on how to support the patient provider-information technology partnership to provide more patient-centered health care. They recommended that AMIA take an active leadership role in consumer health informatics. Specific recommendations were made concerning research, new patient record systems, provider support, information access and evaluation, and policy and regulation. PMID- 11418538 TI - Medical subject headings used to search the biomedical literature. AB - The National Library of Medicine's MEDLINE (MEDLARS Online) database was the first database to be searched nationwide via value-added telecommunication networks. Now available on the World Wide Web free of charge from the National Library of Medicine and from many other sources, it is the world's most heavily used medical database. MEDLINE is unique in that each reference to the medical literature is indexed under a controlled vocabulary called Medical Subject Headings (MeSH). These headings are the keys that unlock the medical literature. MeSH multiplies the usefulness of the MEDLINE database and makes it possible to search the medical literature as we do today. This paper commemorates the 40th anniversary of the introduction of MeSH and salutes some of the farsighted persons who conceived and developed the MEDLINE database. PMID- 11418539 TI - A primer on aspects of cognition for medical informatics. AB - As a multidisciplinary field, medical informatics draws on a range of disciplines, such as computer science, information science, and the social and cognitive sciences. The cognitive sciences can provide important insights into the nature of the processes involved in human- computer interaction and help improve the design of medical information systems by providing insight into the roles that knowledge, memory, and strategies play in a variety of cognitive activities. In this paper, the authors survey literature on aspects of medical cognition and provide a set of claims that they consider to be important in medical informatics. PMID- 11418540 TI - Expanding the guidelines for electronic communication with patients: application to a specific tool. AB - In 1998, the American Medical Informatics Association (AMIA) published a white paper entitled "Guidelines for the Clinical Use of Electronic Mail with Patients," which outlined a practical framework for this interaction. Interest in the use of other Internet-based tools, such as the World Wide Web, to enhance clinical communication is increasing. In such systems, static information can be made centrally available to patients and interactive tools such as messaging systems, schedules, and individualized care regimens can be integrated within the site. Site-specific guidelines are needed to address potential problems inherent in the particular services being offered. This article presents advice on developing site-specific guidelines, with examples, based on experience gained in developing and refining guidelines for the use of PatientWeb at the Massachusetts General Hospital Department of Neurology. PMID- 11418541 TI - A manifesto on telehealth and telemedicine. AB - An evident contrast exists between the generally easy way medical doctors and administrators use the term "telemedicine" and the wide variety of significantly different technologic methods and devices necessary for correctly performing specific tasks in the field. Many misunderstandings could be avoided by agreeing on the types of services that telemedicine can provide, names for those services, and descriptions of what is included in the services. This manifesto lists representative services, with a proposed name for and description of each. PMID- 11418542 TI - A UMLS-based knowledge acquisition tool for rule-based clinical decision support system development. AB - Decision support systems in the medical field have to be easily modified by medical experts themselves. The authors have designed a knowledge acquisition tool to facilitate the creation and maintenance of a knowledge base by the domain expert and its sharing and reuse by other institutions. The Unified Medical Language System (UMLS) contains the domain entities and constitutes the relations repository from which the expert builds, through a specific browser, the explicit domain ontology. The expert is then guided in creating the knowledge base according to the pre-established domain ontology and condition-action rule templates that are well adapted to several clinical decision-making processes. Corresponding medical logic modules are eventually generated. The application of this knowledge acquisition tool to the construction of a decision support system in blood transfusion demonstrates the value of such a pragmatic methodology for the design of rule-based clinical systems that rely on the highly progressive knowledge embedded in hospital information systems. PMID- 11418543 TI - Controlled trial of direct physician order entry: effects on physicians' time utilization in ambulatory primary care internal medicine practices. AB - OBJECTIVE: Direct physician order entry (POE) offers many potential benefits, but evidence suggests that POE requires substantially more time than traditional paper-based ordering methods. The Medical Gopher is a well-accepted system for direct POE that has been in use for more than 15 years. The authors hypothesized that physicians using the Gopher would not spend any more time writing orders than physicians using paper-based methods. DESIGN: A randomized controlled trial of POE using the Medical Gopher system in 11 primary care internal medicine practices. MEASUREMENTS: The authors collected detailed time use data using time motion studies of the physicians and surveyed their opinions about the POE system. RESULTS: The authors found that physicians using the Gopher spent 2.2 min more per patient overall, but when duplicative and administrative tasks were taken into account, physicians were found to have spent only 0.43 min more per patient. With experience, the order entry time fell by 3.73 min per patient. The survey revealed that the physicians believed that the system improved their patient care and wanted the Gopher to continue to be available in their practices. CONCLUSIONS: Little extra time, if any, was required for physicians to use the POE system. With experience in its use, physicians may even save time while enjoying the many benefits of POE. PMID- 11418544 TI - Electronic messaging between primary and secondary care: a four-year case report. AB - OBJECTIVE: To observe how electronic messaging between a hospital consultant and general practitioners (GPs) in 15 practices about patients suffering from diabetes evolved over a 3-year period after an initial 1-year study. DESIGN: Case report. Electronic messages between a hospital consultant and GPs were counted. The authors determined whether a message sent by the consultant was integrated into the receiving GP's electronic medical record system. After the observation period, the GPs answered a questionnaire. MEASUREMENTS: The number of electronic messages and the percentage of messages integrated into the electronic medical record. RESULTS: The volume of messages was maintained during the 3 years after the original study. In the original study, the percentage of the messages integrated by the GPs increased during the year. After that study, however, seven GPs stopped integrating data from messages. The extent to which received messages were integrated varied widely among practices. CONCLUSION: The authors conclude that extrapolation of the results of the original study would have led to incorrect conclusions. Although the volume of messages remained stable after the original study, GPs changed their method of handling messages. Initially, all GPs used the opportunity to copy data from the messages into their own records. At the end of the observation period (that is, the 3 years after completion of the original study), more than 50 percent of GPs had ceased copying data from the messages into their own records. The majority of GPs, however, wanted to expand the use of electronic messaging. PMID- 11418545 TI - Derivation and evaluation of a document-naming nomenclature. AB - OBJECTIVE: The Computerized Patient Record System is deployed at all 173 Veterans Affairs (VA) medical centers. Providers access clinical notes in the system from a note title menu. Following its implementation at the Nashville VA Medical Center, users expressed dissatisfaction with the time required find notes among hundreds of irregularly structured titles. The authors' objective was to develop a document-naming nomenclature (DNN) that creates informative, structured note titles that improve information access. DESIGN: One thousand ninety-four unique note titles from two VA medical centers were reviewed. A note-naming nomenclature and compositional syntax were derived. Compositional order was determined by user preference survey. MEASUREMENTS: The DNN was evaluated by modeling note titles from the Salt Lake City VA Medical Center (n=877), Vanderbilt University Medical Center (n=554), and the Mayo Clinic (n=42). A preliminary usability evaluation was conducted on a structured title display and sorting application. RESULTS: Classes of note title components were found by inspection. Components describe characteristics of the author, the health care event, and the organizational unit providing care. Terms were taken from VA medical center information systems and national standards. The DNN model accurately described 97 to 99 percent of note titles from the test sites. The DNN term coverage varied, depending on component and site. Users found the DNN title format useful and the DNN-based title sorting and note review application easy to learn and quick to use. CONCLUSION: The DNN accurately models note titles at five medical centers. Preliminary usability data indicate that DNN integration with title parsing and sorting software enhances information access. PMID- 11418546 TI - Searching for clinical prediction rules in MEDLINE. AB - OBJECTIVES: Clinical prediction rules have been advocated as a possible mechanism to enhance clinical judgment in diagnostic, therapeutic, and prognostic assessment. Despite renewed interest in the their use, inconsistent terminology makes them difficult to index and retrieve by computerized search systems. No validated approaches to locating clinical prediction rules appear in the literature. The objective of this study was to derive and validate an optimal search filter for retrieving clinical prediction rules, using the National Library of Medicine's MEDLINE database. DESIGN: A comparative, retrospective analysis was conducted. The "gold standard" was established by a manual search of all articles from select print journals for the years 1991 through 1998, which identified articles covering various aspects of clinical prediction rules such as derivation, validation, and evaluation. Search filters were derived, from the articles in the July through December issues of the journals (derivation set), by analyzing the textwords (words in the title and abstract) and the medical subject heading (from the MeSH Thesaurus) used to index each article. The accuracy of these filters in retrieving clinical prediction rules was then assessed using articles in the January through June issues (validation set). MEASUREMENTS: The sensitivity, specificity, positive predictive value, and positive likelihood ratio of several different search filters were measured. RESULTS: The filter "predict$ OR clinical$ OR outcome$ OR risk$" retrieved 98 percent of clinical prediction rules. Four filters, such as "predict$ OR validat$ OR rule$ OR predictive value of tests," had both sensitivity and specificity above 90 percent. The top-performing filter for positive predictive value and positive likelihood ratio in the validation set was "predict$.ti. AND rule$." CONCLUSIONS: Several filters with high retrieval value were found. Depending on the goals and time constraints of the searcher, one of these filters could be used. PMID- 11418547 TI - Information systems can prevent errors and improve quality. PMID- 11418548 TI - Dual regulatory control of a particle maturation function of bacteriophage P1. AB - A unique arrangement of promoter elements was found upstream of the bacteriophage P1 particle maturation gene (mat). A P1-specific late-promoter sequence with conserved elements located at positions -22 and -10 was expected from the function of the gene in phage morphogenesis. In addition to a late-promoter sequence, a -35 element and an operator sequence for the major repressor protein, C1, were found. The -35 and -10 elements constituted an active Escherichia coli sigma(70) consensus promoter, which was converted into a P1-regulated early promoter by the superimposition of a C1 operator. This combination of early- and late-promoter elements regulates and fine-tunes the expression of the particle maturation gene. During lysogenic growth the gene is turned off by P1 immunity functions. Upon induction of lytic growth, the expression of mat starts simultaneously with the expression of other C1-regulated P1 early functions. However, while most of the latter functions are downregulated during late stages of lytic growth the expression of mat continues throughout the entire lytic growth cycle of bacteriophage P1. Thus, the maturation function has a head start on the structural components of the phage particle. PMID- 11418549 TI - Energy-dependent conformational change in the TolA protein of Escherichia coli involves its N-terminal domain, TolQ, and TolR. AB - TolQ, TolR, and TolA inner membrane proteins of Escherichia coli are involved in maintaining the stability of the outer membrane. They share homology with the ExbB, ExbD, and TonB proteins, respectively. The last is involved in energy transduction between the inner and the outer membrane, and its conformation has been shown to depend on the presence of the proton motive force (PMF), ExbB, and ExbD. Using limited proteolysis experiments, we investigated whether the conformation of TolA was also affected by the PMF. We found that dissipation of the PMF by uncouplers led to the formation of a proteinase K digestion fragment of TolA not seen when uncouplers are omitted. This fragment was also detected in Delta tolQ, Delta tolR, and tolA(H22P) mutants but, in contrast to the parental strain, was also seen in the absence of uncouplers. We repeated those experiments in outer membrane mutants such as lpp, pal, and Delta rfa mutants: the behavior of TolA in lpp mutants was similar to that observed with the parental strain. However, the proteinase K-resistant fragment was never detected in the Delta rfa mutant. Altogether, these results suggest that TolA is able to undergo a PMF dependent change of conformation. This change requires TolQ, TolR, and a functional TolA N-terminal domain. The potential role of this energy-dependent process in the stability of the outer membrane is discussed. PMID- 11418550 TI - Constitutive septal murein synthesis in Escherichia coli with impaired activity of the morphogenetic proteins RodA and penicillin-binding protein 2. AB - The pattern of peptidoglycan (murein) segregation in cells of Escherichia coli with impaired activity of the morphogenetic proteins penicillin-binding protein 2 and RodA has been investigated by the D-cysteine-biotin immunolabeling technique (M. A. de Pedro, J. C. Quintela, J.-V. Holtje, and H. Schwarz, J. Bacteriol. 179:2823-2834, 1997). Inactivation of these proteins either by amdinocillin treatment or by mutations in the corresponding genes, pbpA and rodA, respectively, leads to the generation of round, osmotically stable cells. In normal rod-shaped cells, new murein precursors are incorporated all over the lateral wall in a diffuse manner, being mixed up homogeneously with preexisting material, except during septation, when strictly localized murein synthesis occurs. In contrast, in rounded cells, incorporation of new precursors is apparently a zonal process, localized at positions at which division had previously taken place. Consequently, there is no mixing of new and old murein. Old murein is preserved for long periods of time in large, well-defined areas. We propose that the observed patterns are the result of a failure to switch off septal murein synthesis at the end of septation events. Furthermore, the segregation results confirm that round cells of rodA mutants do divide in alternate, perpendicular planes as previously proposed (K. J. Begg and W. D. Donachie, J. Bacteriol. 180:2564-2567, 1998). PMID- 11418551 TI - Mutations in genes involved in the flagellar export apparatus of the solvent tolerant Pseudomonas putida DOT-T1E strain impair motility and lead to hypersensitivity to toluene shocks. AB - Pseudomonas putida DOT-T1E is a solvent-tolerant strain able to grow in the presence of 1% (vol/vol) toluene in the culture medium. Random mutagenesis with mini-Tn5-'phoA-Km allowed us to isolate a mutant strain (DOT-T1E-42) that formed blue colonies on Luria-Bertani medium supplemented with 5-bromo-4-chloro-3 indolylphosphate and that, in contrast to the wild-type strain, was unable to tolerate toluene shocks (0.3%, vol/vol). The mutant strain exhibited patterns of tolerance or sensitivity to a number of antibiotics, detergents, and chelating agents similar to those of the wild-type strain. The mutation in this strain therefore seemed to specifically affect toluene tolerance. Cloning and sequencing of the mutation revealed that the mini-Tn5-'phoA-Km was inserted within the fliP gene, which is part of the fliLMNOPQRflhBA cluster, a set of genes that encode flagellar structure components. FliP is involved in the export of flagellar proteins, and in fact, the P. putida fliP mutant was nonmotile. The finding that, after replacing the mutant allele with the wild-type one, the strain recovered the wild-type pattern of toluene tolerance and motility unequivocally assigned FliP a function in solvent resistance. An flhB knockout mutant, another gene component of the flagellar export apparatus, was also nonmotile and hypersensitive to toluene. In contrast, a nonpolar mutation at the fliL gene, which encodes a cytoplasmic membrane protein associated with the flagellar basal body, yielded a nonmotile yet toluene-resistant strain. The results are discussed regarding a possible role of the flagellar export apparatus in the transport of one or more proteins necessary for toluene tolerance in P. putida DOT-T1E to the periplasm. PMID- 11418552 TI - OhrR is a repressor of ohrA, a key organic hydroperoxide resistance determinant in Bacillus subtilis. AB - Bacillus subtilis displays a complex adaptive response to the presence of reactive oxygen species. To date, most proteins that protect against reactive oxygen species are members of the peroxide-inducible PerR and sigma(B) regulons. We investigated the function of two B. subtilis homologs of the Xanthomonas campestris organic hydroperoxide resistance (ohr) gene. Mutational analyses indicate that both ohrA and ohrB contribute to organic peroxide resistance in B. subtilis, with the OhrA protein playing the more important role in growing cells. Expression of ohrA, but not ohrB, is strongly and specifically induced by organic peroxides. Regulation of ohrA requires the convergently transcribed gene, ohrR, which encodes a member of the MarR family of transcriptional repressors. In an ohrR mutant, ohrA expression is constitutive, whereas expression of the neighboring ohrB gene is unaffected. Selection for mutant strains that are derepressed for ohrA transcription identifies a perfect inverted repeat sequence that is required for OhrR-mediated regulation and likely defines an OhrR binding site. Thus, B. subtilis contains at least three regulons (sigma(B), PerR, and OhrR) that contribute to peroxide stress responses. PMID- 11418553 TI - Sodium ion-driven serine/threonine transport in Porphyromonas gingivalis. AB - Porphyromonas gingivalis is an asaccharolytic, gram-negative bacterium that relies on the fermentation of amino acids for metabolic energy. When grown in continuous culture in complex medium containing 4 mM (each) free serine, threonine, and arginine, P. gingivalis assimilated mainly glutamate/glutamine, serine, threonine, aspartate/asparagine, and leucine in free and/or peptide form. Serine and threonine were assimilated in approximately equal amounts in free and peptide form. We characterized serine transport in this bacterium by measuring uptake of the radiolabeled amino acid in washed cells of P. gingivalis energized with a tetrapeptide not containing serine. Serine was transported by a single system with an affinity constant for transport (K(t)) of 24 microM that was competitively inhibited by threonine. Serine transport was dependent on sodium ion concentration in the suspending buffer, and the addition of the ionophore gramicidin caused the inhibition of serine uptake. Together these data indicate that serine transport was sodium ion-motive force driven. A P. gingivalis gene potentially encoding a serine transporter was identified by sequence similarity to an Escherichia coli serine transporter (SstT). This P. gingivalis gene, designated sstT, was inactivated by insertion of a Bacteroides tetQ gene, producing the mutant W50ST. The mutant was unable to transport serine, confirming the presence of a single serine transporter in this bacterium under these growth conditions. The transport of serine by P. gingivalis was dependent on the presence of free cysteine in the suspension buffer. Other reducing agents were unable to stimulate serine uptake. These data show that P. gingivalis assimilates free serine and threonine from culture media via a cysteine-activated, sodium ion motive force-driven serine/threonine transporter. PMID- 11418554 TI - Purification, characterization, and genetic analysis of Cu-containing dissimilatory nitrite reductase from a denitrifying halophilic archaeon, Haloarcula marismortui. AB - Cu-containing dissimilatory nitrite reductase (CuNiR) was purified from denitrifying cells of a halophilic archaeon, Haloarcula marismortui. The purified CuNiR appeared blue in the oxidized state, possessing absorption peaks at 600 and 465 nm in the visible region. Electron paramagnetic resonance spectroscopy suggested the presence of type 1 Cu (g(II) = 2.232; A(II) = 4.4 mT) and type 2 Cu centers (g(II) = 2.304; A(II) = 13.3 mT) in the enzyme. The enzyme contained two subunits, whose apparent molecular masses were 46 and 42 kDa, according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. N-terminal amino acid sequence analysis indicated that the two subunits were identical, except that the 46-kDa subunit was 16 amino acid residues longer than the 42-kDa subunit in the N terminal region. A nirK gene encoding the CuNiR was cloned and sequenced, and the deduced amino acid sequence with a residual length of 361 amino acids was homologous (30 to 41%) with bacterial counterparts. Cu-liganding residues His 133, Cys-174, His-182, and Met-187 (for type 1 Cu) and His-138, His-173, and His 332 (for type 2 Cu) were conserved in the enzyme. As generally observed in the halobacterial enzymes, the enzymatic activity of the purified CuNiR was enhanced during increasing salt concentration and reached its maximum in the presence of 2 M NaCl with the value of 960 microM NO(2)(-) x min(-1) x mg(-1). PMID- 11418555 TI - Design of a protein-targeting system for lactic acid bacteria. AB - We designed an expression and export system that enabled the targeting of a reporter protein (the staphylococcal nuclease Nuc) to specific locations in Lactococcus lactis cells, i.e., cytoplasm, cell wall, or medium. Optimization of protein secretion and of protein cell wall anchoring was performed with L. lactis cells by modifying the signals located at the N and C termini, respectively, of the reporter protein. Efficient translocation of precursor (approximately 95%) is obtained using the signal peptide from the lactococcal Usp45 protein and provided that the mature protein is fused to overall anionic amino acids at its N terminus; those residues prevented interactions of Nuc with the cell envelope. Nuc could be covalently anchored to the peptidoglycan by using the cell wall anchor motif of the Streptococcus pyogenes M6 protein. However, the anchoring step proved to not be totally efficient in L. lactis, as considerable amounts of protein remained membrane associated. Our results may suggest that the defect is due to limiting sortase in the cell. The optimized expression and export vectors also allowed secretion and cell wall anchoring of Nuc in food-fermenting and commensal strains of Lactobacillus. In all strains tested, both secreted and cell wall-anchored Nuc was enzymatically active, suggesting proper enzyme folding in the different locations. These results provide the first report of a targeting system in lactic acid bacteria in which the final location of a protein is controlled and biological activity is maintained. PMID- 11418556 TI - Cloning and characterization of the Flavobacterium johnsoniae gliding motility genes gldD and gldE. AB - Cells of Flavobacterium johnsoniae move over surfaces by a process known as gliding motility. The mechanism of this form of motility is not known. Cells of F. johnsoniae propel latex spheres along their surfaces, which is thought to be a manifestation of the motility machinery. Three of the genes that are required for F. johnsoniae gliding motility, gldA, gldB, and ftsX, have recently been described. Tn4351 mutagenesis was used to identify another gene, gldD, that is needed for gliding. Tn4351-induced gldD mutants formed nonspreading colonies, and cells failed to glide. They also lacked the ability to propel latex spheres and were resistant to bacteriophages that infect wild-type cells. Introduction of wild-type gldD into the mutants restored motility, ability to propel latex spheres, and sensitivity to bacteriophage infection. gldD codes for a cytoplasmic membrane protein that does not exhibit strong sequence similarity to proteins of known function. gldE, which lies immediately upstream of gldD, encodes another cytoplasmic membrane protein that may be involved in gliding motility. Overexpression of gldE partially suppressed the motility defects of a gldB point mutant, suggesting that GldB and GldE may interact. GldE exhibits sequence similarity to Borrelia burgdorferi TlyC and Salmonella enterica serovar Typhimurium CorC. PMID- 11418557 TI - The SHI-3 iron transport island of Shigella boydii 0-1392 carries the genes for aerobactin synthesis and transport. AB - In Shigella boydii 0-1392, genes encoding the synthesis and transport of the hydroxamate siderophore aerobactin are located within a 21-kb iron transport island between lysU and the pheU tRNA gene. DNA sequence analysis of the S. boydii 0-1392 island, designated SHI-3 for Shigella island 3, revealed a conserved aerobactin operon associated with a P4 prophage-like integrase gene and numerous insertion sequences (IS). SHI-3 is present at the pheU tRNA locus in some S. boydii isolates but not in others. The map locations of the aerobactin genes vary among closely related species. The association of the aerobactin operon with phage genes and mobile elements and its presence at different locations within the genomes of enteric pathogens suggest that these virulence enhancing genes may have been acquired by bacteriophage integration or IS element mediated transposition. An S. boydii aerobactin synthesis mutant, 0-1392 iucB, was constructed and was similar to the wild type in tissue culture assays of invasion and intercellular spread. PMID- 11418558 TI - Identification of a DNA binding region in GerE from Bacillus subtilis. AB - Proteins that have a structure similar to those of LuxR and FixJ comprise a large subfamily of transcriptional activator proteins. Most members of the LuxR-FixJ family contain a similar amino-terminal receiver domain linked by a small region to a carboxy-terminal domain that contains an amino acid sequence similar to the helix-turn-helix (HTH) motif found in other DNA-binding proteins. GerE from Bacillus subtilis is the smallest member of the LuxR-FixJ family. Its 74-amino acid sequence is similar over its entire length to the DNA binding region of this protein family, including the HTH motif. Therefore, GerE provides a simple model for studies of the role of this HTH domain in DNA binding. Toward this aim, we sought to identify the amino acids within this motif that are important for the specificity of binding to DNA. We examined the effects of single base pair substitutions in the high-affinity GerE binding site on the sigK promoter and found that nucleotides at positions +2, +3, and +4 relative to the transcription start site on the sigK promoter are important for a high-affinity interaction with GerE. We next examined the effects of single alanine substitutions at two positions in the HTH region of GerE on binding to wild-type or mutant target sites. We found that the substitution of an alanine for the threonine at position 42 of GerE produced a protein that binds with equal affinity to two sites that differ by 1 bp, whereas wild-type GerE binds with different affinities to these two sites. These results provide evidence that the amino acyl residues in or near the putative HTH region of GerE and potentially other members of the LuxR-FixJ family determine the specificity of DNA binding. PMID- 11418559 TI - Control of the arabinose regulon in Bacillus subtilis by AraR in vivo: crucial roles of operators, cooperativity, and DNA looping. AB - The proteins involved in the utilization of L-arabinose by Bacillus subtilis are encoded by the araABDLMNPQ-abfA metabolic operon and by the araE/araR divergent unit. Transcription from the ara operon, araE transport gene, and araR regulatory gene is induced by L-arabinose and negatively controlled by AraR. The purified AraR protein binds cooperatively to two in-phase operators within the araABDLMNPQ abfA (OR(A1) and OR(A2)) and araE (OR(E1) and OR(E2)) promoters and noncooperatively to a single operator in the araR (OR(R3)) promoter region. Here, we have investigated how AraR controls transcription from the ara regulon in vivo. A deletion analysis of the ara promoters region showed that the five AraR binding sites are the key cis-acting regulatory elements of their corresponding genes. Furthermore, OR(E1)-OR(E2) and OR(R3) are auxiliary operators for the autoregulation of araR and the repression of araE, respectively. Analysis of mutations designed to prevent cooperative binding of AraR showed that in vivo repression of the ara operon requires communication between repressor molecules bound to two properly spaced operators. This communication implicates the formation of a small loop by the intervening DNA. In an in vitro transcription system, AraR alone sufficed to abolish transcription from the araABDLMNPQ-abfA operon and araE promoters, strongly suggesting that it is the major protein involved in the repression mechanism of L-arabinose-inducible expression in vivo. The ara regulon is an example of how the architecture of the promoters is adapted to respond to the particular characteristics of the system, resulting in a tight and flexible control. PMID- 11418560 TI - The alkane hydroxylase gene of Burkholderia cepacia RR10 is under catabolite repression control. AB - In many microorganisms the first step for alkane degradation is the terminal oxidation of the molecule by an alkane hydroxylase. We report the characterization of a gene coding for an alkane hydroxylase in a Burkholderia cepacia strain isolated from an oil-contaminated site. The protein encoded showed similarity to other known or predicted bacterial alkane hydroxylases, although it clustered on a separate branch together with the predicted alkane hydroxylase of a Mycobacterium tuberculosis strain. Introduction of the cloned B. cepacia gene into an alkane hydroxylase knockout mutant of Pseudomonas fluorescens CHAO restored its ability to grow on alkanes, which confirms that the gene analyzed encodes a functional alkane hydroxylase. The gene, which was named alkB, is not linked to other genes of the alkane oxidation pathway. Its promoter was identified, and its expression was analyzed under different growth conditions. Transcription was induced by alkanes of chain lengths containing 12 to at least 30 carbon atoms as well as by alkanols. Although the gene was efficiently expressed during exponential growth, transcription increased about fivefold when cells approached stationary phase, a characteristic not shared by the few alkane degraders whose regulation has been studied. Expression of the alkB gene was under carbon catabolite repression when cells were cultured in the presence of several organic acids and sugars or in a complex (rich) medium. The catabolic repression process showed several characteristics that are clearly different from what has been observed in other alkane degradation pathways. PMID- 11418561 TI - Indole can act as an extracellular signal in Escherichia coli. AB - Previous work has shown that lacZ fusions to the cysK, astD, tnaB, and gabT genes in Escherichia coli are activated by self-produced extracellular signals. Using a combination of ethyl acetate extraction, reversed-phase C(18) chromatography, and thin-layer chromatography, we have purified an extracellular activating signal from E. coli supernatants. Mass spectrometry revealed a molecule with an m/z peak of 117, consistent with indole. Nuclear magnetic resonance analysis of the purified E. coli factor and synthetic indole revealed identical profiles. Using synthetic indole, a dose-dependent activation was observed with lacZ fusions to the gabT, astD, and tnaB genes. However, cysK::lacZ and several control fusions were not significantly activated by indole. Conditioned medium prepared from a tnaA (tryptophanase) mutant, deficient in indole production, supported 26 to 41% lower activation of the gabT and astD fusions. The residual level of activation may be due to a second activating signal. Activation of the tnaB::lacZ fusion was reduced by greater than 70% in conditioned medium from a tnaA mutant. PMID- 11418562 TI - Accumulation of the PhaP phasin of Ralstonia eutropha is dependent on production of polyhydroxybutyrate in cells. AB - Polyhydroxyalkanoates (PHAs) are polyoxoesters that are produced by diverse bacteria and that accumulate as intracellular granules. Phasins are granule associated proteins that accumulate to high levels in strains that are producing PHAs. The accumulation of phasins has been proposed to be dependent on PHA production, a model which is now rigorously tested for the phasin PhaP of Ralstonia eutropha. R. eutropha phaC PHA synthase and phaP phasin gene replacement strains were constructed. The strains were engineered to express heterologous and/or mutant PHA synthase alleles and a phaP-gfp translational fusion in place of the wild-type alleles of phaC and phaP. The strains were analyzed with respect to production of polyhydroxybutyrate (PHB), accumulation of PhaP, and expression of the phaP-gfp fusion. The results suggest that accumulation of PhaP is strictly dependent on the genetic capacity of strains to produce PHB, that PhaP accumulation is regulated at the level of both PhaP synthesis and PhaP degradation, and that, within mixed populations of cells, PhaP accumulation within cells of a given strain is not influenced by PHB production in cells of other strains. Interestingly, either the synthesis of PHB or the presence of relatively large amounts of PHB in cells (>50% of cell dry weight) is sufficient to enable PhaP synthesis. The results suggest that R. eutropha has evolved a regulatory mechanism that can detect the synthesis and presence of PHB in cells and that PhaP expression can be used as a marker for the production of PHB in individual cells. PMID- 11418563 TI - PhcS represses gratuitous expression of phenol-metabolizing enzymes in Comamonas testosteroni R5. AB - We identified an open reading frame, designated phcS, downstream of the transcriptional activator gene (phcR) for the expression of multicomponent phenol hydroxylase (mPH) in Comamonas testosteroni R5. The deduced product of phcS was homologous to AphS of C. testosteroni TA441, which belongs to the GntR family of transcriptional regulators. The transformation of Pseudomonas aeruginosa PAO1c (phenol negative, catechol positive) with pROR502 containing phcR and the mPH genes conferred the ability to grow on phenol, while transformation with pROR504 containing phcS, phcR, and mPH genes did not confer this ability. The disruption of phcS in strain R5 had no effect on its phenol-oxygenating activity in a chemostat culture with phenol. The phenol-oxygenating activity was not expressed in strain R5 grown in a chemostat with acetate. In contrast, the phenol oxygenating activity in the strain with a knockout phcS gene when grown in a chemostat with acetate as the limiting growth factor was 66% of that obtained in phenol-grown cells of the strain with a knockout in the phcS gene. The disruption of phcS and/or phcR and the complementation in trans of these defects confirm that PhcS is a trans-acting repressor and that the unfavorable expression of mPH in the phcS knockout cells grown on acetate requires PhcR. These results show that the PhcS protein repressed the gratuitous expression of phenol-metabolizing enzymes in the absence of the genuine substrate and that strain R5 acted by an unknown mechanism in which the PhcS-mediated repression was overcome in the presence of the pathway substrate. PMID- 11418564 TI - PhaC and PhaR are required for polyhydroxyalkanoic acid synthase activity in Bacillus megaterium. AB - Polyhydroxyalkanoic acids (PHAs) are a class of polyesters stored in inclusion bodies and found in many bacteria and in some archaea. The terminal step in the synthesis of PHA is catalyzed by PHA synthase. Genes encoding this enzyme have been cloned, and the primary sequence of the protein, PhaC, is deduced from the nucleotide sequences of more than 30 organisms. PHA synthases are grouped into three classes based on substrate range, molecular mass, and whether or not there is a requirement for phaE in addition to the phaC gene product. Here we report the results of an analysis of a PHA synthase that does not fit any of the described classes. This novel PHA synthase from Bacillus megaterium required PhaC (PhaC(Bm)) and PhaR (PhaR(Bm)) for activity in vivo and in vitro. PhaC(Bm) showed greatest similarity to the PhaCs of class III in both size and sequence. Unlike those in class III, the 40-kDa PhaE was not required, and furthermore, the 22-kDa PhaR(Bm) had no obvious homology to PhaE. Previously we showed that PhaC(Bm), and here we show that PhaR(Bm), is localized to inclusion bodies in living cells. We show that two forms of PHA synthase exist, an active form in PHA-accumulating cells and an inactive form in nonaccumulating cells. PhaC was constitutively produced in both cell types but was more susceptible to protease degradation in the latter type. Our data show that the role of PhaR is posttranscriptional and that it functions directly or indirectly with PhaC(Bm) to produce an active PHA synthase. PMID- 11418565 TI - Role of arginines in coenzyme A binding and catalysis by the phosphotransacetylase from Methanosarcina thermophila. AB - Phosphotransacetylase (EC 2.3.1.8) catalyzes the reversible transfer of the acetyl group from acetyl phosphate to coenzyme A (CoA): CH(3)COOPO(3)(2-) + CoASH <==> CH(3)COSCoA + HPO(4)(2-). The role of arginine residues was investigated for the phosphotransacetylase from Methanosarcina thermophila. Kinetic analysis of a suite of variants indicated that Arg 87 and Arg 133 interact with the substrate CoA. Arg 87 variants were reduced in the ability to discriminate between CoA and the CoA analog 3'-dephospho-CoA, indicating that Arg 87 forms a salt bridge with the 3'-phosphate of CoA. Arg 133 is postulated to interact with the 5'-phosphate of CoA. Large decreases in k(cat) and k(cat)/K(m) for all of the Arg 87 and Arg 133 variants indicated that these residues are also important, although not essential, for catalysis. Large decreases in k(cat) and k(cat)/K(m) were also observed for the variants in which lysine replaced Arg 87 and Arg 133, suggesting that the bidentate interaction of these residues with CoA or their greater bulk is important for optimal activity. Desulfo-CoA is a strong competitive inhibitor of the enzyme, suggesting that the sulfhydryl group of CoA is important for the optimization of CoA-binding energy but not for tight substrate binding. Chemical modification of the wild-type enzyme by 2,3-butanedione and substrate protection by CoA indicated that at least one reactive arginine is in the active site and is important for activity. The inhibition pattern of the R87Q variant indicated that Arg 87 is modified, which contributes to the inactivation; however, at least one additional active-site arginine is modified leading to enzyme inactivation, albeit at a lower rate. PMID- 11418566 TI - Succinate dehydrogenase and other respiratory pathways in thylakoid membranes of Synechocystis sp. strain PCC 6803: capacity comparisons and physiological function. AB - Respiration in cyanobacterial thylakoid membranes is interwoven with photosynthetic processes. We have constructed a range of mutants that are impaired in several combinations of respiratory and photosynthetic electron transport complexes and have examined the relative effects on the redox state of the plastoquinone (PQ) pool by using a quinone electrode. Succinate dehydrogenase has a major effect on the PQ redox poise, as mutants lacking this enzyme showed a much more oxidized PQ pool. Mutants lacking type I and II NAD(P)H dehydrogenases also had more oxidized PQ pools. However, in the mutant lacking type I NADPH dehydrogenase, succinate was essentially absent and effective respiratory electron donation to the PQ pool could be established after addition of 1 mM succinate. Therefore, lack of the type I NADPH dehydrogenase had an indirect effect on the PQ pool redox state. The electron donation capacity of succinate dehydrogenase was found to be an order of magnitude larger than that of type I and II NAD(P)H dehydrogenases. The reason for the oxidized PQ pool upon inactivation of type II NADH dehydrogenase may be related to the facts that the NAD pool in the cell is much smaller than that of NADP and that the NAD pool is fully reduced in the mutant without type II NADH dehydrogenase, thus causing regulatory inhibition. The results indicate that succinate dehydrogenase is the main respiratory electron transfer pathway into the PQ pool and that type I and II NAD(P)H dehydrogenases regulate the reduction level of NADP and NAD, which, in turn, affects respiratory electron flow through succinate dehydrogenase. PMID- 11418567 TI - Characterization of an aminoacylase from the hyperthermophilic archaeon Pyrococcus furiosus. AB - Aminoacylase was identified in cell extracts of the hyperthermophilic archaeon Pyrococcus furiosus by its ability to hydrolyze N-acetyl-L-methionine and was purified by multistep chromatography. The enzyme is a homotetramer (42.06 kDa per subunit) and, as purified, contains 1.0 +/- 0.48 g-atoms of zinc per subunit. Treatment of the purified enzyme with EDTA resulted in complete loss of activity. This was restored to 86% of the original value (200 U/mg) by treatment with ZnCl(2) (and to 74% by the addition of CoCl(2)). After reconstitution with ZnCl(2), the enzyme contained 2.85 +/- 0.48 g-atoms of zinc per subunit. Aminoacylase showed broad substrate specificity and hydrolyzed nonpolar N acylated L amino acids (Met, Ala, Val, and Leu), as well as N-formyl-L methionine. The high K(m) values for these compounds indicate that the enzyme plays a role in the metabolism of protein growth substrates rather than in the degradation of cellular proteins. Maximal aminoacylase activity with N-acetyl-L methionine as the substrate occurred at pH 6.5 and a temperature of 100 degrees C. The N-terminal amino acid sequence of the purified aminoacylase was used to identify, in the P. furiosus genome database, a gene that encodes 383 amino acids. The gene was cloned and expressed in Escherichia coli by using two approaches. One involved the T7 lac promoter system, in which the recombinant protein was expressed as inclusion bodies. The second approach used the Trx fusion system, and this produced soluble but inactive recombinant protein. Renaturation and reconstitution experiments with Zn(2+) ions failed to produce catalytically active protein. A survey of databases showed that, in general, organisms that contain a homolog of the P. furiosus aminoacylase (> or = 50% sequence identity) utilize peptide growth substrates, whereas those that do not contain the enzyme are not known to be proteolytic, suggesting a role for the enzyme in primary catabolism. PMID- 11418568 TI - trans-3-Chloroacrylic acid dehalogenase from Pseudomonas pavonaceae 170 shares structural and mechanistic similarities with 4-oxalocrotonate tautomerase. AB - The genes (caaD1 and caaD2) encoding the trans-3-chloroacrylic acid dehalogenase (CaaD) of the 1,3-dichloropropene-utilizing bacterium Pseudomonas pavonaceae 170 were cloned and heterologously expressed in Escherichia coli and Pseudomonas sp. strain GJ1. CaaD is a protein of 50 kDa that is composed of alpha-subunits of 75 amino acid residues and beta-subunits of 70 residues. It catalyzes the hydrolytic cleavage of the beta-vinylic carbon-chlorine bond in trans-3-chloroacrylic acid with a turnover number of 6.4 s(-1). On the basis of sequence similarity, oligomeric structure, and subunit size, CaaD appears to be related to 4 oxalocrotonate tautomerase (4-OT). This tautomerase consists of six identical subunits of 62 amino acid residues and catalyzes the isomerization of 2-oxo-4 hexene-1,6-dioate, via hydroxymuconate, to yield 2-oxo-3-hexene-1,6-dioate. In view of the oligomeric architecture of 4-OT, a trimer of homodimers, CaaD is postulated to be a hexameric protein that functions as a trimer of alpha beta dimers. The sequence conservation between CaaD and 4-OT and site-directed mutagenesis experiments suggested that Pro-1 of the beta-subunit and Arg-11 of the alpha-subunit are active-site residues in CaaD. Pro-1 could act as the proton acceptor/donor, and Arg-11 is probably involved in carboxylate binding. Based on these findings, a novel dehalogenation mechanism is proposed for the CaaD catalyzed reaction which does not involve the formation of a covalent enzyme substrate intermediate. PMID- 11418569 TI - The Bordetella bhu locus is required for heme iron utilization. AB - Bordetella pertussis and Bordetella bronchiseptica are capable of obtaining iron from hemin and hemoglobin. Genes encoding a putative bacterial heme iron acquisition system (bhu, for Bordetella heme utilization) were identified in a B. pertussis genomic sequence database, and the corresponding DNA was isolated from a virulent strain of B. pertussis. A B. pertussis bhuR mutant, predicted to lack the heme outer membrane receptor, was generated by allelic exchange. In contrast to the wild-type strain, bhuR mutant PM5 was incapable of acquiring iron from hemin and hemoglobin; genetic complementation of PM5 with the cloned bhuRSTUV genes restored heme utilization to wild-type levels. In parallel studies, B. bronchiseptica bhu sequences were also identified and a B. bronchiseptica bhuR mutant was constructed and confirmed to be defective in heme iron acquisition. The wild-type B. bronchiseptica parent strain grown under low-iron conditions produced the presumptive BhuR protein, which was absent in the bhuR mutant. Furthermore, production of BhuR by iron-starved B. bronchiseptica was markedly enhanced by culture in hemin-supplemented medium, suggesting that these organisms sense and respond to heme in the environment. Analysis of the genetic region upstream of the bhu cluster identified open reading frames predicted to encode homologs of the Escherichia coli ferric citrate uptake regulators FecI and FecR. These putative Bordetella regulators may mediate heme-responsive positive transcriptional control of the bhu genes. PMID- 11418570 TI - Succinyl-CoA:(R)-benzylsuccinate CoA-transferase: an enzyme of the anaerobic toluene catabolic pathway in denitrifying bacteria. AB - Anaerobic microbial toluene catabolism is initiated by addition of fumarate to the methyl group of toluene, yielding (R)-benzylsuccinate as first intermediate, which is further metabolized via beta-oxidation to benzoyl-coenzyme A (CoA) and succinyl-CoA. A specific succinyl-CoA:(R)-benzylsuccinate CoA-transferase activating (R)-benzylsuccinate to the CoA-thioester was purified and characterized from Thauera aromatica. The enzyme is fully reversible and forms exclusively the 2-(R)-benzylsuccinyl-CoA isomer. Only some close chemical analogs of the substrates are accepted by the enzyme: succinate was partially replaced by maleate or methylsuccinate, and (R)-benzylsuccinate was replaced by methylsuccinate, benzylmalonate, or phenylsuccinate. In contrast to all other known CoA-transferases, the enzyme consists of two subunits of similar amino acid sequences and similar sizes (44 and 45 kDa) in an alpha(2)beta(2) conformation. Identity of the subunits with the products of the previously identified toluene induced bbsEF genes was confirmed by determination of the exact masses via electrospray-mass spectrometry. The deduced amino acid sequences resemble those of only two other characterized CoA-transferases, oxalyl-CoA:formate CoA transferase and (E)-cinnamoyl-CoA:(R)-phenyllactate CoA-transferase, which represent a new family of CoA-transferases. As suggested by kinetic analysis, the reaction mechanism of enzymes of this family apparently involves formation of a ternary complex between the enzyme and the two substrates. PMID- 11418571 TI - Structural and functional characterization of IS679 and IS66-family elements. AB - A new insertion sequence (IS) element, IS679 (2,704 bp in length), has been identified in plasmid pB171 of enteropathogenic Escherichia coli B171. IS679 has imperfect 25-bp terminal inverted repeats (IRs) and three open reading frames (ORFs) (here called tnpA, tnpB, and tnpC). A plasmid carrying a composite transposon (Tn679) with the kanamycin resistance gene flanked by an intact IS679 sequence and an IS679 fragment with only IRR (IR on the right) was constructed to clarify the transposition activity of IS679. A transposition assay done with a mating system showed that Tn679 could transpose at a high frequency to the F plasmid derivative used as the target. On transposition, Tn679 duplicated an 8-bp sequence at the target site. Tn679 derivatives with a deletion in each ORF of IS679 did not transpose, finding indicative that all three IS679 ORFs are essential for transposition. The tnpA and tnpC products appear to have the amino acid sequence motif characteristic of most transposases. A homology search of the databases found that a total of 25 elements homologous to IS679 are present in Agrobacterium, Escherichia, Rhizobium, Pseudomonas, and Vibrio spp., providing evidence that the elements are widespread in gram-negative bacteria. We found that these elements belong to the IS66 family, as do other elements, including nine not previously reported. Almost all of the elements have IRs similar to those in IS679 and, like IS679, most appear to have duplicated an 8-bp sequence at the target site on transposition. These elements have three ORFs corresponding to those in IS679, but many have a mutation(s) in an ORF(s). In almost all of the elements, tnpB is located in the -1 frame relative to tnpA, such that the initiation codon of tnpB overlaps the TGA termination codon of tnpA. In contrast, tnpC, separated from tnpB by a space of ca. 20 bp, is located in any one of three frames relative to tnpB. No common structural features were found around the intergenic regions, indicating that the three ORFs are expressed by translational coupling but not by translational frameshifting. PMID- 11418572 TI - Autotrophic CO(2) fixation by Chloroflexus aurantiacus: study of glyoxylate formation and assimilation via the 3-hydroxypropionate cycle. AB - In the facultative autotrophic organism Chloroflexus aurantiacus, a phototrophic green nonsulfur bacterium, the Calvin cycle does not appear to be operative in autotrophic carbon assimilation. An alternative cyclic pathway, the 3 hydroxypropionate cycle, has been proposed. In this pathway, acetyl coenzyme A (acetyl-CoA) is assumed to be converted to malate, and two CO(2) molecules are thereby fixed. Malyl-CoA is supposed to be cleaved to acetyl-CoA, the starting molecule, and glyoxylate, the carbon fixation product. Malyl-CoA cleavage is shown here to be catalyzed by malyl-CoA lyase; this enzyme activity is induced severalfold in autotrophically grown cells. Malate is converted to malyl-CoA via an inducible CoA transferase with succinyl-CoA as a CoA donor. Some enzyme activities involved in the conversion of malonyl-CoA via 3-hydroxypropionate to propionyl-CoA are also induced under autotrophic growth conditions. So far, no clue as to the first step in glyoxylate assimilation has been obtained. One possibility for the assimilation of glyoxylate involves the conversion of glyoxylate to glycine and the subsequent assimilation of glycine. However, such a pathway does not occur, as shown by labeling of whole cells with [1,2 (13)C(2)]glycine. Glycine carbon was incorporated only into glycine, serine, and compounds that contained C(1) units derived therefrom and not into other cell compounds. PMID- 11418573 TI - Localization of GerAA and GerAC germination proteins in the Bacillus subtilis spore. AB - The GerAA, -AB, and -AC proteins of the Bacillus subtilis spore are required for the germination response to L-alanine as the sole germinant. They are likely to encode the components of the germination apparatus that respond directly to this germinant, mediating the spore's response; multiple homologues of the gerA genes are found in every spore former so far examined. The gerA operon is expressed in the forespore, and the level of expression of the operon appears to be low. The GerA proteins are predicted to be membrane associated. In an attempt to localize GerA proteins, spores of B. subtilis were broken and fractionated to give integument, membrane, and soluble fractions. Using antibodies that detect Ger proteins specifically, as confirmed by the analysis of strains lacking GerA and the related GerB proteins, the GerAA protein and the GerAC+GerBC protein homologues were localized to the membrane fraction of fragmented spores. The spore-specific penicillin-binding protein PBP5*, a marker for the outer forespore membrane, was absent from this fraction. Extraction of spores to remove coat layers did not release the GerAC or AA protein from the spores. Both experimental approaches suggest that GerAA and GerAC proteins are located in the inner spore membrane, which forms a boundary around the cellular compartment of the spore. The results provide support for a model of germination in which, in order to initiate germination, germinant has to permeate the coat and cortex of the spore and bind to a germination receptor located in the inner membrane. PMID- 11418574 TI - ISAfe1, an ISL3 family insertion sequence from Acidithiobacillus ferrooxidans ATCC 19859. AB - A 1.3-kb insertion sequence, termed ISAfe1 (U66426), from Acidithiobacillus ferrooxidans ATCC 19859 is described. ISAfe1 exhibits the features of a typical bacterial insertion sequence. It has 26-bp, imperfectly matched, terminal inverted repeats and an open reading frame (ORF) that potentially encodes a transposase (TPase) of 404 amino acids (AAB07489) with significant similarity to members of the ISL3 family of insertion sequences. A potential ribosome-binding site and potential -10 and -35 promoter sites for the TPase ORF were identified, and a +1 transcriptional start site was detected experimentally. A potential outwardly directed -35 site was identified in the right inverted repeat of ISAfe1. A second ORF (ORF B), of unknown function, was found on the complementary strand with significant similarity to ORF 2 of ISAe1 from Ralstonia eutropha. Southern blot analyses demonstrated that ISAfe1-like elements can be found in multiple copies in a variety of A. ferrooxidans strains and that they exhibit transposition. A codon adaptation index (CAI) analysis of the TPase of ISAfe1 indicates that is has a CAI of 0.726 and can be considered well adapted to its host, suggesting that ISAfe1 might be an ancient resident of A. ferrooxidans. Analysis of six of its target sites of insertion in the genome of A. ferrooxidans ATCC 19859 indicates a preference for 8-bp pseudopalindromic sequences, one of which resembles the termini of its inverted repeats. Evidence is presented here that is consistent with the possibility that ISAfe1 can promote both plasmid cointegrate formation and resolution in E. coli. PMID- 11418575 TI - Multiple domains are required for the toxic activity of Pseudomonas aeruginosa ExoU. AB - Expression of ExoU by Pseudomonas aeruginosa is correlated with acute cytotoxicity in a number of epithelial and macrophage cell lines. In vivo, ExoU is responsible for epithelial injury. The absence of a known motif or significant homology with other proteins suggests that ExoU may possess a new mechanism of toxicity. To study the intracellular effects of ExoU, we developed a transient transfection system in Chinese hamster ovary cells. Transfection with full-length but not truncated forms of ExoU inhibited reporter gene expression. Inhibition of reporter activity after cotransfection with ExoU-encoding constructs was correlated with cellular permeability and death. The toxicity of truncated versions of ExoU could be restored by coexpression of the remainder of the molecule from separate plasmids in trans. This strategy was used to map N- and C terminal regions of ExoU that are necessary but not sufficient for toxicity. Disruption of a middle region of the protein reduces toxicity. This portion of the molecule is postulated to allow the N- and C-terminal regions to functionally complement one another. In contrast to ExoS and ExoT, native and recombinant ExoU molecules do not oligomerize or form aggregates. The complex domain structure of ExoU suggests that, like other P. aeruginosa-encoded type III effectors (ExoS and ExoT), ExoU toxicity may result from a molecule that possesses more than one activity. PMID- 11418576 TI - Identification and distribution of new insertion sequences in the genome of alkaliphilic Bacillus halodurans C-125. AB - Fifteen kinds of new insertion sequences (ISs), IS641 to IS643, IS650 to IS658, IS660, IS662, and IS663, and a group II intron (Bh.Int) were identified in the 4,202,352-bp genome of alkaliphilic Bacillus halodurans C-125. Out of 120 ISs identified in the C-125 genome, 29 were truncated, indicating the occurrence of internal rearrangements of the genome. The ISs other than IS650, IS653, IS660, and IS663 generated a 2- to 9-bp duplication of the target site sequence, and the ISs other than IS650, IS653, and IS657 carry 14- to 64-bp inverted repeats. Sequence analysis revealed that six kinds of ISs (IS642, IS643, IS654, IS655, IS657, and IS658) belong to a separate IS family (IS630, IS21, IS256, IS3, IS200/IS605, and IS30, respectively) as a new member. Also, IS651 and IS652 were characterized as new members of the ISL3 family. Significant similarity was found between the transposase (Tpase) sequences between IS650 and IS653 (78.2%), IS651 and IS652 (56.3%), IS656 and IS662 (71.0%), and IS660 and IS663 (44.5%), but the others showed no similarity to one another. Tpases in 28 members of IS651 in the C-125 genome were found to have become diversified. Most of the IS elements widely distributed throughout the genome were inserted in noncoding regions, although some genes, such as those coding for an ATP-binding cassette transporter/permease, a response regulator, and L-indole 2-dehydrogenase, have been mutated through the insertion of IS elements. It is evident, however, that not all IS elements have transposed and caused rearrangements of the genome in the past 17 years during which strain C-125 was subcultured under neutral and alkaline conditions. PMID- 11418577 TI - Gene replacement analysis of the butyrolactone autoregulator receptor (FarA) reveals that FarA acts as a Novel regulator in secondary metabolism of Streptomyces lavendulae FRI-5. AB - IM-2 [(2R,3R,1'R)-2-1'-hydroxybutyl-3-hydroxymethyl gamma-butanolide] is a gamma butyrolactone autoregulator which, in Streptomyces lavendulae FRI-5, switches off the production of D-cycloserine but switches on the production of a blue pigment and several nucleoside antibiotics. To clarify the in vivo function of an IM-2 specific receptor (FarA) in the IM-2 signaling cascade of S. lavendulae FRI-5, a farA deletion mutant was constructed by means of homologous recombination. On several solid media, no significant difference in morphology was observed between the wild-type strain and the farA mutant (strain K104), which demonstrated that the IM-2-FarA system does not participate in the morphological control of S. lavendulae FRI-5. In liquid media, the farA mutant overproduced nucleoside antibiotics and produced blue pigment earlier than did the wild-type strain, suggesting that the FarA protein acts primarily as a negative regulator on the biosynthesis of these compounds in the absence of IM-2. However, contrary to the IM-2-dependent suppression of D-cycloserine production in the wild-type strain, overproduction of D-cycloserine was observed in the farA mutant, indicating for the first time that the presence of both IM-2 and intact FarA are necessary for the suppression of D-cycloserine biosynthesis. PMID- 11418578 TI - The PDZ domain of the SpoIVB serine peptidase facilitates multiple functions. AB - During spore formation in Bacillus subtilis, the SpoIVB protein is a critical component of the sigma(K) regulatory checkpoint. SpoIVB has been shown to be a serine peptidase that is synthesized in the spore chamber and which self-cleaves, releasing active forms. These forms can signal proteolytic processing of the transcription factor sigma(K) in the outer mother cell chamber of the sporulating cell. This forms the basis of the sigma(K) checkpoint and ensures accurate sigma(K)-controlled gene expression. SpoIVB has also been shown to activate a second distinct process, termed the second function, which is essential for the formation of heat-resistant spores. In addition to the serine peptidase domain, SpoIVB contains a PDZ domain. We have altered a number of conserved residues in the PDZ domain by site-directed mutagenesis and assayed the sporulation phenotype and signaling properties of mutant SpoIVB proteins. Our work has revealed that the SpoIVB PDZ domain could be used for up to four distinct processes, (i) targeting of itself for trans proteolysis, (ii) binding to the protease inhibitor BofC, (iii) signaling of pro-sigma(K) processing, and (iv) signaling of the second function of SpoIVB. PMID- 11418579 TI - Evidence that an additional mutation is required to tolerate insertional inactivation of the Streptomyces lividans recA gene. AB - In contrast to recA of other bacteria, the recA gene of Streptomyces lividans has been described as indispensable for viability (G. Muth, D. Frese, A. Kleber, and W. Wohlleben, Mol. Gen. Genet. 255:420-428, 1997.). Therefore, a closer analysis of this gene was performed to detect possible unique features distinguishing the Streptomyces RecA protein from the well-characterized Escherichia coli RecA protein. The S. lividans recA gene restored UV resistance and recombination activity of an E. coli recA mutant. Also, transcriptional regulation was similar to that of E. coli recA. Gel retardation experiments showed that S. lividans recA is also under control of the Streptomyces SOS repressor LexA. The S. lividans recA gene could be replaced only by simultaneously expressing a plasmid encoded recA copy. Surprisingly, the recA expression plasmid could subsequently be eliminated using an incompatible plasmid without the loss of viability. Besides being UV sensitive and recombination deficient, all the mutants were blocked in sporulation. Genetic complementation restored UV resistance and recombination activity but did not affect the sporulation defect. This indicated that all the recA mutants had suffered from an additional mutation, which might allow toleration of a recA deficiency. PMID- 11418580 TI - Spontaneous erythromycin resistance mutation in a 23S rRNA gene, rrlA, of the extreme thermophile Thermus thermophilus IB-21. AB - Spontaneous, erythromycin-resistant mutants of Thermus thermophilus IB-21 were isolated and found to carry the mutation A2058G in one of two 23S rRNA operons. The heterozygosity of these mutants indicates that A2058G confers a dominant or codominant phenotype in this organism. This mutation provides a valuable tool for the genetic manipulation of the 23S rRNA genes of Thermus. PMID- 11418581 TI - New plasmid system to select for Saccharomyces cerevisiae purine-cytosine permease affinity mutants. AB - The FCY2 gene of Saccharomyces cerevisiae encodes a purine-cytosine permease (PCP) that mediates the active transport of purines and cytosine. A structure function model for this PCP has been recently proposed. In this study, we developed a plasmid-based system that generated a number of affinity-mutated alleles, enabling us to define new amino acids critical for permease function. PMID- 11418582 TI - Multiple genes for the last step of proline biosynthesis in Bacillus subtilis. AB - The complete Bacillus subtilis genome contains four genes (proG, proH, proI, and comER) with the potential to encode Delta(1)-pyrroline-5-carboxylate reductase, a proline biosynthetic enzyme. Simultaneous defects in three of these genes (proG, proH, and proI) were required to confer proline auxotrophy, indicating that the products of these genes are mostly interchangeable with respect to the last step in proline biosynthesis. PMID- 11418583 TI - Design and properties of N(CCG)-gp41, a chimeric gp41 molecule with nanomolar HIV fusion inhibitory activity. AB - The design and characterization of a chimeric protein, termed N(CCG)-gp41, derived from the ectodomain of human immunodeficiency virus (HIV), type I gp41 is described. N(CCG)-gp41 features an exposed trimeric coiled-coil comprising the N terminal helices of the gp41 ectodomain. The trimeric coiled-coil is stabilized both by fusion to a minimal thermostable ectodomain of gp41 and by engineered intersubunit disulfide bonds. N(CCG)-gp41 is shown to inhibit HIV envelope mediated cell fusion at nanomolar concentrations with an IC(50) of 16.1 +/- 2.8 nm. It is proposed that N(CCG)-gp41 targets the exposed C-terminal region of the gp41 ectodomain in its pre-hairpin intermediate state, thereby preventing the formation of the fusogenic form of the gp41 ectodomain, which comprises a highly stable trimer of hairpins arranged in a six-helix bundle. N(CCG)-gp41 has potential as a therapeutic agent for the direct inhibition of HIV cell entry, as an anti-HIV vaccine, and as a component of a rapid throughput assay for screening for small molecule inhibitors of HIV envelope-mediated cell fusion. It is anticipated that antibodies raised against N(CCG)-gp41 may target the trimeric coiled-coil of N-terminal helices of the gp41 ectodomain that is exposed in the pre-hairpin intermediate state in a manner analogous to peptides derived from the C-terminal helix of gp41 that are currently in clinical trials. PMID- 11418584 TI - Expression and characterization of a murine enzyme able to cleave beta-carotene. The formation of retinoids. AB - Because animals are not able to synthesize retinoids de novo, ultimately they must derive them from dietary provitamin A carotenoids through a process known as carotene cleavage. The enzyme responsible for catalyzing carotene cleavage (beta carotene 15,15'-dioxygenase) has been characterized primarily in rat intestinal scrapings. Using a recently reported cDNA sequence for a carotene cleavage enzyme from Drosophila, we identified a cDNA encoding a mouse homolog of this enzyme. When the cDNA was expressed in either Escherichia coli or Chinese hamster ovary cells, expression conferred upon bacterial and Chinese hamster ovary cell homogenates the ability to cleave beta-carotene to retinal. Several lines of evidence obtained upon kinetic analyses of the recombinant enzyme suggested that carotene cleavage enzyme interacts with other proteins present within cell or tissue homogenates. This was confirmed by pull-down experiments upon incubation of recombinant enzyme with tissue 12,000 x g supernatants. Matrix-assisted laser desorption ionization-mass spectrometry analysis of pulled-down proteins indicates that an atypical testis-specific isoform of lactate dehydrogenase associates with recombinant carotene cleavage enzyme. mRNA transcripts for the carotene cleavage enzyme were detected by reverse transcription-polymerase chain reaction in mouse testes, liver, kidney, and intestine. In situ hybridization studies demonstrated that carotene cleavage enzyme is expressed prominently in maternal tissue surrounding the embryo but not in embryonic tissues at 7.5 and 8.5 days postcoitus. This work offers new insights for understanding the biochemistry of carotene cleavage to retinoids. PMID- 11418585 TI - Occurrence of oligosialic acids on integrin alpha 5 subunit and their involvement in cell adhesion to fibronectin. AB - Integrin alpha(5)beta(1), a major fibronectin receptor, functions in a wide variety of biological phenomena. We have found that alpha 2-8-linked oligosialic acids with 5 < or = degree of polymerization (DP) < or = 7 occur on integrin alpha(5) subunit of the human melanoma cell line G361. The integrin alpha(5) subunit immunoprecipitated with anti-integrin alpha(5) antibody reacted with the monoclonal antibody 12E3, which recognizes oligo/polysialic acid with DP > or = 5 but not with the polyclonal antibody H.46 recognizing oligo/polysialic acid with DP > or = 8. The occurrence of oligosialic acids was further demonstrated by fluorometric C(7)/C(9) analysis on the immunopurified integrin alpha(5) subunit. Oligosialic acids were also found in the alpha(5) subunit of several other human cells such as foreskin fibroblast and chronic erythroleukemia K562 cells. These results suggest the ubiquitous modification with unique oligosialic acids occurs on the alpha(5) subunit of integrin alpha(5)beta(1). The adhesion of human melanoma G361 cells to fibronectin was mainly mediated by integrin alpha(5)beta(1). Treatment of cells with sialidase from Arthrobacter ureafaciens cleaving alpha 2-3-, alpha 2-6-, and alpha 2-8-linked sialic acids inhibited adhesion to fibronectin. On the other hand, N-acetylneuraminidase II, which cleaves alpha 2-3 and alpha 2-6 but not alpha 2-8 linkages, showed no inhibitory activity. After the loss of oligosialic acids, integrin alpha(5)beta(1) failed to bind to fibronectin-conjugated Sepharose, indicating that the oligosialic acid on the alpha(5) subunit of integrin alpha(5)beta(1) plays important roles in cell adhesion to fibronectin. PMID- 11418586 TI - Stathmin inhibition enhances okadaic acid-induced mitotic arrest: a potential role for stathmin in mitotic exit. AB - Stathmin is a microtubule-destabilizing phosphoprotein that plays a critical role in the regulation of mitosis. The microtubule-depolymerizing activity of stathmin is lost upon phosphorylation in mitosis. Although the role of phosphorylation of stathmin by p34(cdc2) kinase in the assembly of the mitotic spindle is well established, the role of dephosphorylation of stathmin in mitosis is unknown. In this study, we tested the hypothesis that dephosphorylation of stathmin may be critically important for the depolymerization of the mitotic spindle and the exit from mitosis. We compared the effects of okadaic acid, a specific inhibitor of serine/threonine protein phosphatases, on different parameters of mitotic progression in the presence or absence of stathmin deficiency. Because okadaic acid prevents dephosphorylation of stathmin and results in accumulation of the inactive phosphorylated form, exposure to okadaic acid would be expected to have a more profound effect on mitosis in the presence of relative stathmin deficiency. We found that inhibition of stathmin expression results in increased sensitivity to the antimitotic effects of okadaic acid. This was reflected by increased growth inhibition associated with mitotic arrest. A vast majority of the stathmin-inhibited cells were found to be arrested in late metaphase/anaphase and had severe mitotic spindle abnormalities. Exposure to okadaic acid also resulted in a bigger ratio of polymerized/unpolymerized tubulin in stathmin inhibited cells relative to control cells. Because the only difference between the control and the stathmin-inhibited cells is the deficiency of stathmin in the latter, the increased susceptibility of the stathmin-inhibited cells to okadaic acid-induced mitotic arrest implies a role for stathmin in the later stages of mitosis. PMID- 11418587 TI - Impaired synergistic activation of stress-activated protein kinase SAPK/JNK in mouse embryonic stem cells lacking SEK1/MKK4: different contribution of SEK2/MKK7 isoforms to the synergistic activation. AB - Stress-activated protein kinase/c-Jun NH(2)-terminal kinase (SAPK/JNK), which is a member of the mitogen-activated protein kinase (MAPK) family, plays an important role in a stress-induced signaling cascade. SAPK/JNK activation requires the phosphorylation of Thr and Tyr residues in its Thr-Pro-Tyr motif, and SEK1 (MKK4) and MKK7 (SEK2) have been identified as the upstream MAPK kinases. Here we examined the activation and phosphorylation sites of SAPK/JNK and differentiated the contribution of SEK1 and MKK7alpha1, -gamma1, and -gamma2 isoforms to the MAPK activation. In SEK1-deficient mouse embryonic stem cells, stress-induced SAPK/JNK activation was markedly impaired, and this defect was accompanied with a decreased level of the Tyr phosphorylation. Analysis in HeLa cells co-transfected with the two MAPK kinases revealed that the Thr and Tyr of SAPK/JNK were independently phosphorylated in response to heat shock by MKK7gamma1 and SEK1, respectively. However, MKK7alpha1 failed to phosphorylate the Thr of SAPK/JNK unless its Tyr residue was phosphorylated by SEK1. In contrast, MKK7gamma2 had the ability to phosphorylate both Thr and Tyr residues. In all cases, the dual phosphorylation of the Thr and Tyr residues was essentially required for the full activation of SAPK/JNK. These data provide the first evidence that synergistic activation of SAPK/JNK requires both phosphorylation at the Thr and Tyr residues in living cells and that the preference for the Thr and Tyr phosphorylation was different among the members of MAPK kinases. PMID- 11418588 TI - Modulation of the helicase activity of eIF4A by eIF4B, eIF4H, and eIF4F. AB - Eukaryotic initiation factor (eIF) 4A is a DEAD box RNA helicase that works in conjunction with eIF4B, eIF4H, or as a subunit of eIF4F to unwind secondary structure in the 5'-untranslated region of mRNA, which facilitates binding of the mRNA to the 40 S ribosomal subunit. This study demonstrates how the helicase activity of eIF4A is modulated by eIF4B, eIF4H, or as a subunit of eIF4F. Results indicate that a linear relationship exists between the initial rate or amplitude of unwinding and duplex stability for all factor combinations tested. eIF4F, like eIF4A, behaves as a non-processive helicase. Either eIF4B or eIF4H stimulated the initial rate and amplitude of eIF4A-dependent duplex unwinding, and the magnitude of stimulation is dependent on duplex stability. Furthermore, eIF4A (or eIF4F) becomes a slightly processive helicase in the presence of eIF4B or eIF4H. All combinations of factors tested indicate that the rate of duplex unwinding is equivalent in the 5' --> 3' and 3' --> 5' directions. However, the optimal rate of unwinding was dependent on the length of the single-stranded region of the substrate when different combinations of factors were used. The combinations of eIF4A, eIF4A + eIF4B, eIF4A + eIF4H, and eIF4F showed differences in their ability to unwind chemically modified duplexes. A simple model of how eIF4B or eIF4H affects the duplex unwinding mechanism of eIF4A is proposed. PMID- 11418589 TI - Coupling between cyclooxygenase, terminal prostanoid synthase, and phospholipase A2. AB - We have recently shown that two distinct prostaglandin (PG) E(2) synthases show preferential functional coupling with upstream cyclooxygenase (COX)-1 and COX-2 in PGE(2) biosynthesis. To investigate whether other lineage-specific PG synthases also show preferential coupling with either COX isozyme, we introduced these enzymes alone or in combination into 293 cells to reconstitute their functional interrelationship. As did the membrane-bound PGE(2) synthase, the perinuclear enzymes thromboxane synthase and PGI(2) synthase generated their respective products via COX-2 in preference to COX-1 in both the -induced immediate and interleukin-1-induced delayed responses. Hematopoietic PGD(2) synthase preferentially used COX-1 and COX-2 in the -induced immediate and interleukin-1-induced delayed PGD(2)-biosynthetic responses, respectively. This enzyme underwent stimulus-dependent translocation from the cytosol to perinuclear compartments, where COX-1 or COX-2 exists. COX selectivity of these lineage specific PG synthases was also significantly affected by the concentrations of arachidonate, which was added exogenously to the cells or supplied endogenously by the action of cytosolic or secretory phospholipase A(2). Collectively, the efficiency of coupling between COXs and specific PG synthases may be crucially influenced by their spatial and temporal compartmentalization and by the amount of arachidonate supplied by PLA(2)s at a moment when PG production takes place. PMID- 11418590 TI - A novel myocyte-specific gene Midori promotes the differentiation of P19CL6 cells into cardiomyocytes. AB - Although several cardiac-specific transcription factors have been shown to play vital roles in various steps during the heart formation, the precise mechanism of the early stage of cardiogenesis has yet to be elucidated. By differential display technique, we tried to identify molecules that are expressed earlier than cardiac transcription factors such as CSX/NKX2-5 and GATA-4 and are involved in cardiomyocyte differentiation using the P19CL6 cell line, which efficiently differentiates into cardiomyocytes when treated with dimethyl sulfoxide. We isolated a novel gene designated Midori. Its deduced amino acid sequence contained an ATP/GTP-binding site, Ig-like domain, and Kringle-like domain. Northern blot analysis revealed that expression of Midori was restricted to the fetal and adult heart and adult skeletal muscle in mice. In whole mount in situ hybridization, Midori was expressed in cardiac crescent and developing heart but not in somites. The MIDORI protein was localized in the nucleus and overexpression of Midori induced expression of endogenous Midori itself, suggesting that MIDORI may act as a transcriptional regulator. Permanent P19CL6 cell lines overexpressing Midori more efficiently differentiated into cardiomyocytes than did parental cells, whereas those overexpressing the antisense Midori less efficiently differentiated. These results suggest that Midori may promote the differentiation of P19CL6 into cardiomyocytes. PMID- 11418591 TI - The impact of N-glycosylation on the functions of polysialyltransferases. AB - Poly-alpha-2,8-sialic acid (polysialic acid) is a post-translational modification of the neural cell adhesion molecule (NCAM) and an important regulator of neuronal cell-cell interactions. The synthesis of polysialic acid depends on the two polysialyltransferases ST8SiaII and ST8SiaIV. Understanding the catalytic mechanisms of the polysialyltransferases is critical toward the aim of influencing physiological and pathophysiological functions mediated by polysialic acid. We recently demonstrated that polysialyltransferases are bifunctional enzymes exhibiting auto- and NCAM polysialylation activity. Autopolysialylation occurs on N-glycans of the enzymes, and glycosylation variants lacking sialic acid and galactose were found to be inactive for both auto- and NCAM polysialylation. In the present study, we have analyzed the number and functional importance of N-linked oligosaccharides present on polysialyltransferases. We demonstrate that autopolysialylation depends on specific N-glycans attached to Asn(74) in ST8SiaIV and Asn(89) and Asn(219) in ST8SiaII. Deletion of polysialic acid acceptor sites by site-directed mutagenesis rendered the polysialyltransferases inactive in vitro and in vivo. The inactivity of autopolysialylation-negative polysialyltransferases in vivo was not caused by the absence or default targeting of the enzymes. The data presented in this study clearly show that active polysialyltransferases are competent to perform autopolysialylation and provide strong evidence for a tight functional link between the two catalytic functions. PMID- 11418592 TI - Interactions between adaptor protein-1 of the clathrin coat and microtubules via type 1a microtubule-associated proteins. AB - The classical view suggests that adaptor proteins of the clathrin coat mediate the sorting of cargo protein passengers into clathrin-coated pits and the recruitment of clathrin into budding areas in the donor membrane. In the present study, we provide biochemical and morphological evidence that the adaptor protein 1 (AP-1) adaptor of the trans-Golgi network clathrin interacts with microtubules. AP-1 in cytosolic extracts interacted with in vitro assembled microtubules, and these interactions were inhibited by ATP depletion of the extracts or in the presence of 5'-adenylylimidodiphosphate. An overexpressed gamma-subunit of the AP 1 complex associated with microtubules, suggesting that this subunit may mediate the interaction of AP-1 with the cytoskeleton. Purified AP-1 did not interact with purified microtubules, but interaction occurred when an isolated microtubule associated protein fraction was added to the reaction mix. The gamma-adaptin subunit of AP-1 specifically co-immunoprecipitated with a microtubule-associated protein of type 1a from rat brain cytosol. This suggests that type 1a microtubule associated protein may mediate the association of AP-1 with microtubules in the cytoplasm. The microtubule binding activity of AP-1 was markedly inhibited in cytosol of mitotic cells. By means of its interaction with microtubule-associated proteins, we propose novel roles for AP-1 adaptors in modulating the dynamics of the cytoskeleton, the stability and shape of coated organelles, and the loading of nascent AP-1-coated vesicles onto appropriate microtubular tracks. PMID- 11418593 TI - Dopamine beta-monooxygenase signal/anchor sequence alters trafficking of peptidylglycine alpha-hydroxylating monooxygenase. AB - Dopamine beta-monooxygenase (DBM) and peptidylglycine alpha-hydroxylating monooxygenase (PHM) are essential for the biosynthesis of catecholamines and amidated peptides, respectively. The enzymes share a conserved catalytic core. We studied the role of the DBM signal sequence by appending it to soluble PHM (PHMs) and expressing the DBMsignal/PHMs chimera in AtT-20 and Chinese hamster ovary cells. PHMs produced as part of DBMsignal/PHMs was active. In vitro translated and cellular DBMsignal/PHMs had similar masses, indicating that the DBM signal was not removed. DBMsignal/PHMs was membrane-associated and had the properties of an intrinsic membrane protein. After in vitro translation in the presence of microsomal membranes, trypsin treatment removed 2 kDa from DBMsignal/PHMs while PHMs was entirely protected. In addition, a Cys residue in DBMsignal/PHMs was accessible to Cys-directed biotinylation. Thus the chimera adopts the topology of a type II membrane protein. Pulse-chase experiments indicate that DBMsignal/PHMs turns over rapidly after exiting the trans-Golgi network. Although PHMs is efficiently localized to secretory granules, DBMsignal/PHMs is largely localized to the endoplasmic reticulum in AtT-20 cells. On the basis of stimulated secretion, the small amount of PHMs generated is stored in secretory granules. In contrast, the expression of DBMsignal/PHMs in PC12 cells yields protein that is localized to secretory granules. PMID- 11418594 TI - The p42/p44 mitogen-activated protein kinase activation triggers p27Kip1 degradation independently of CDK2/cyclin E in NIH 3T3 cells. AB - The p42/p44 mitogen-activated protein (MAP) kinase is stimulated by various mitogenic stimuli, and its sustained activation is necessary for cell cycle G(1) progression and G(1)/S transition. G(1) progression and G(1)/S transition also depend on sequential cyclin-dependent kinase (CDK) activation. Here, we demonstrate that MAP kinase inhibition leads to accumulation of the CDK inhibitor p27(Kip1) in NIH 3T3 cells. Blocking the proteasome-dependent degradation of p27(Kip1) impaired this accumulation, suggesting that MAP kinase does not act on p27(Kip1) protein synthesis. In the absence of extracellular signals (growth factors or cell adhesion), genetic activation of MAP kinase decreased the expression of p27(Kip1) as assessed by cotransfection experiments and by immunofluorescence detection. Importantly, MAP kinase activation also decreased the expression of a p27(Kip1) mutant, which cannot be phosphorylated by CDK2, suggesting that MAP kinase-dependent p27(Kip1) regulation is CDK2-independent. Accordingly, expression of dominant-negative CDK2 did not impair the down regulation of p27(Kip1) induced by MAP kinase activation. These data demonstrate that the MAP kinase pathway regulates p27(Kip1) expression in fibroblasts essentially through a degradation mechanism, independently of p27(Kip1) phosphorylation by CDK2. This strengthens the role of this CDK inhibitor as a key effector of G(1) growth arrest, whose expression can be controlled by extracellular stimuli-dependent signaling pathways. PMID- 11418595 TI - E2F transcriptional activation requires TRRAP and GCN5 cofactors. AB - The E2F family of transcription factors regulates the temporal transcription of genes involved in cell cycle progression and DNA synthesis. E2F transactivation is antagonized by retinoblastoma protein (pRb), which recruits chromatin remodeling proteins such as histone deacetylases and SWI.SNF complexes to the promoter to repress transcription. We hypothesized that E2F proteins must reverse the pRb-imposed chromatin structure to stimulate transcription. If this is true, E2F proteins should recruit proteins capable of histone acetylation. Here we map the E2F-4 transactivation domain and show that E2F-1 and E2F-4 transactivation domains bind the acetyltransferase GCN5 and cofactor TRRAP in vivo. TRRAP and GCN5 co-expression stimulated E2F-mediated transactivation, and c-Myc repressed E2F transactivation dependent on an intact TRRAP/GCN5 binding motif. The transactivation domain of E2F-4 recruited proteins with significant histone acetyltransferase activity in vivo, and this activity required catalytically active GCN5. E2F-4 proteins with subtle mutations in the transactivation domain exhibited a positive correlation among transcriptional activation and GCN5 and TRRAP binding capacity and associated acetyltransferase activity. We conclude that E2F stimulates transcription by recruiting acetyltransferase activity and the essential cofactors GCN5 and TRRAP. These results provide a mechanism for E2F transcription factors to overcome pRb-mediated dominant repression of transcription. PMID- 11418596 TI - The Gal4 activation domain binds Sug2 protein, a proteasome component, in vivo and in vitro. AB - An in vivo protein interaction assay was used to search a yeast cDNA library for proteins that bind to the acidic activation domain (AD) of the yeast Gal4 protein. Sug2 protein, a component of the 19 S regulatory particle of the 26 S proteasome, was one of seven proteins identified in this screen. In vitro binding assays confirm a direct interaction between these proteins. SUG2 and SUG1, another 19 S component, were originally discovered as a mutation able to suppress the phenotype of a Gal4 truncation mutant (Gal4(D)p) lacking much of its AD. Sug1p has previously been shown to bind the Gal4 AD in vitro. Taken together, these genetic and biochemical data suggest a biologically significant interaction between the Gal4 protein and the 19 S regulatory particle of the proteasome. Indeed, it is demonstrated here that the Gal4 AD interacts specifically with immunopurified 19 S complex. The proteasome regulatory particle has been shown recently to play a direct role in RNA polymerase II transcription and the activator-19 S interaction could be important in recruiting this large complex to transcriptionally active GAL genes. PMID- 11418597 TI - Peptide mimics of the vesicular stomatitis virus G-protein transmembrane segment drive membrane fusion in vitro. AB - The efficiency of cell-cell fusion mediated by heterologously expressed vesicular stomatitis virus G-protein has previously been shown to be affected by mutating its transmembrane segment. Here, we show that a synthetic peptide modeled after this transmembrane segment drives liposome-liposome fusion. Addition of millimolar Ca(2+) concentrations strongly potentiated the effect of the peptides suggesting that Ca(2+)-mediated liposome aggregation supports the activity of the peptide. Peptide-driven fusion was suppressed by lysolipid, an established inhibitor of natural membrane fusion, and involved inner and outer leaflets of the liposomal bilayer. Thus, transmembrane segment peptide-driven liposome fusion exhibits important hallmarks characteristic of natural membrane fusion. Importantly, the mutations previously shown to attenuate the function of full length G-protein in cell-cell fusion also attenuated the fusogenicity of the peptide, albeit in a less pronounced fashion. Therefore, the function of the peptide mimic is dependent on its primary structure, similar to full-length G protein. Together, our data suggest that the G-protein transmembrane segment is an autonomous functional domain. We propose that it acts at a late step in membrane fusion elicited by vesicular stomatitis virus. PMID- 11418598 TI - Proteolytic activation of respiratory syncytial virus fusion protein. Cleavage at two furin consensus sequences. AB - The F (fusion) protein of the respiratory syncytial viruses is synthesized as an inactive precursor F(0) that is proteolytically processed at the multibasic sequence KKRKRR(136) into the subunits F(1) and F(2) by the cellular protease furin. This maturation process is essential for the F protein to gain fusion competence. We observed that proteolytic cleavage additionally occurs at another basic motif, RARR(109), that also meets the requirements for furin recognition. Cleavage at both sites leads to the removal from the polypeptide chain of a glycosylated peptide of 27 amino acids. When the sequence RARR(109) was changed to NANR(109) or to RANN(109) by site-directed mutagenesis, cleavage by furin was completely prevented. Although the mutants were still processed at position Arg(136), they did not show any syncytia formation. Proteolytic cleavage of the modified motifs was achieved by treatment of transfected cells with trypsin converting the F mutants into their fusogenic forms. Our findings indicate that both furin consensus sequences have to be cleaved in order to activate the fusion protein. PMID- 11418599 TI - Cofilin phosphorylation and actin reorganization activities of testicular protein kinase 2 and its predominant expression in testicular Sertoli cells. AB - We previously identified testicular protein kinase 1 (TESK1), which phosphorylates cofilin and induces actin cytoskeletal reorganization. We now report identification and characterization of another member of a TESK family, testicular protein kinase 2 (TESK2), with 48% amino acid identity with TESK1. Like TESK1, TESK2 phosphorylated cofilin specifically at Ser-3 and induced formation of actin stress fibers and focal adhesions. Both TESK1 and TESK2 are highly expressed in the testis, but in contrast to TESK1, which is predominantly expressed in testicular germ cells, TESK2 is expressed predominantly in nongerminal Sertoli cells. Thus, TESK1 and TESK2 seem to play distinct roles in spermatogenesis. In HeLa cells, TESK1 was localized mainly in the cytoplasm, whereas TESK2 was localized mainly in the nucleus, which means that TESK1 and TESK2 likely have distinct cellular functions. Because the kinase-inactive mutant of TESK2 was localized in the cytoplasm, nuclear/cytoplasmic localization of TESK2 depends on its kinase activity. A TESK2 mutant lacking the C-terminal noncatalytic region had about a 10-fold higher kinase activity in vitro and, when expressed in HeLa cells, induced punctate actin aggregates in the cytoplasm and unusual condensation and fragmentation of nuclei, followed by apoptosis. Thus, we propose that the C-terminal region plays important roles in regulating the kinase activity and cellular functions of TESK2. PMID- 11418600 TI - The crystal structure of tetanus toxin Hc fragment complexed with a synthetic GT1b analogue suggests cross-linking between ganglioside receptors and the toxin. AB - Tetanus toxin, a member of the family of Clostridial neurotoxins, is one of the most potent toxins known. The crystal structure of the complex of the COOH terminal fragment of the heavy chain with an analogue of its ganglioside receptor, GT1b, provides the first direct identification and characterization of the ganglioside-binding sites. The ganglioside induces cross-linking by binding to two distinct sites on the Hc molecule. The structure sheds new light on the binding of Clostridial neurotoxins to receptors on neuronal cells and provides important information relevant to the design of anti-tetanus and anti-botulism therapeutic agents. PMID- 11418601 TI - Differential gene regulation in human versus rodent hepatocytes by peroxisome proliferator-activated receptor (PPAR) alpha. PPAR alpha fails to induce peroxisome proliferation-associated genes in human cells independently of the level of receptor expresson. AB - We compared the ability of rat and human hepatocytes to respond to fenofibric acid and a novel potent phenylacetic acid peroxisome proliferator-activated receptor (PPAR) alpha agonist (compound 1). Fatty acyl-CoA oxidase (FACO) activity and mRNA were increased after treatment with either fenofibric acid or compound 1 in rat hepatocytes. In addition, apolipoprotein CIII mRNA was decreased by both fenofibric acid and compound 1 in rat hepatocytes. Both agonists decreased apolipoprotein CIII mRNA in human hepatocytes; however, very little change in FACO activity or mRNA was observed. Furthermore, other peroxisome proliferation (PP)-associated genes including peroxisomal 3-oxoacyl CoA thiolase (THIO), peroxisomal enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase (HD), peroxisomal membrane protein-70 (PMP-70) were not regulated by PPAR alpha agonists in human hepatocytes. Moreover, other genes that are regulated by PPAR alpha ligands in human hepatocytes such as mitochondrial HMG CoA synthase and carnitine palmitoyl transferase-1 (CPT-1) were also regulated in HepG2 cells by PPAR alpha agonists. Several stably transfected HepG2 cell lines were established that overexpressed human PPAR alpha to levels between 6- and 26 fold over normal human hepatocytes. These PPAR alpha-overexpressing cells had higher basal mRNA levels of mitochondrial HMG-CoA synthase and CPT-1; however, basal FACO mRNA levels and other PP-associated genes including THIO, HD, or PMP 70 mRNA were not substantially affected. In addition, FACO, THIO, HD, and PMP-70 mRNA levels did not increase in response to PPAR alpha agonist treatment in the PPAR alpha-overexpressing cells, although mitochondrial HMG-CoA synthase and CPT 1 mRNAs were both induced. These results suggest that other factors besides PPAR alpha levels determine the species-specific response of human and rat hepatocytes to the induction of PP. PMID- 11418602 TI - Growth hormone receptor ubiquitination, endocytosis, and degradation are independent of signal transduction via Janus kinase 2. AB - The ubiquitin-proteasome system is required in growth hormone receptor (GHR) endocytosis. For cytokine receptors, which lack intrinsic tyrosine kinase activity, signal transduction is initiated by the activation of a member of the Janus kinase (JAK) family. Previously, we have shown that GHR and JAK2 tyrosine (de) phosphorylation are regulated via the ubiquitin system. In this study, we examined the role of JAK2-mediated signal transduction in GHR internalization and down-regulation. Mutation of the attachment site for JAK2, box-1, in the GHR cytoplasmic tail resulted in the complete absence of GHR and JAK2 phosphorylation. This modification did not alter the rate and extent of receptor bound growth hormone internalization as compared with a functional GHR, nor did it change its turnover and transport to the plasma membrane. In addition, the receptor was still normally ubiquitinated and remained dependent on both an intact ubiquitin system and proteasomal action for its internalization. Thus, GHR ubiquitination, endocytosis, and degradation occur independently of GHR signal transduction via JAK2. We conclude that whereas endocytosis and degradation require the ubiquitin system, they are independent of GHR signal transduction. PMID- 11418603 TI - Metal ion trafficking in earthworms. Identification of a cadmium-specific metallothionein. AB - Exposure to cadmium poses a considerable risk to human health and environmental safety. Earthworms reside in the most contaminated sites on earth, displaying a phenomenal tolerance to toxic heavy metals. They exhibit a distinct metabolic pathway that allows the bio-accumulation of cadmium to yield body burdens in excess of 1/1000th of total dry body weight, a most impressive figure by any standard. However, the precise molecular mechanism underlying this phenomenon remains to be unraveled. This study meets this challenge by fully characterizing the major metal-binding protein in earthworms, namely the two isoforms of metallothionein. Chemical analysis of recombinant protein showed that although both isoforms bind equimolar amounts of cadmium (6 mol), wMT-2 is more stable during proton competition. Furthermore, isoform-specific transcript analysis demonstrated that only wMT-2 is responsive to cadmium in a dose and temporal manner. The specific sequestration of cadmium to wMT-2 protein was confirmed in situ using polyclonal antisera. The latter also provided the means for mapping the cellular and intracellular distribution of metallothionein, thus yielding a holistic insight into its involvement in cadmium transit during absorption, storage, and excretion. The structure-function relationship of wMT-2 and its role in cadmium detoxification through sequestration and compartmentalization is discussed. PMID- 11418604 TI - Cloning of human PRP4 reveals interaction with Clk1. AB - Prp4 is a protein kinase of Schizosaccharomyces pombe identified through its role in pre-mRNA splicing, and belongs to a kinase family including mammalian serine/arginine-rich protein-specific kinases and Clks, whose substrates are serine/arginine-rich proteins. We cloned human PRP4 (hPRP4) full-length cDNA and the antiserum raised against a partial peptide of hPRP4 recognized 170-kDa polypeptide in HeLa S3 cell extracts. Northern blot analysis revealed that hPRP4 mRNA was ubiquitously expressed in multiple tissues. The extended NH(2)-terminal region of hPRP4 contains an arginine/serine-rich domain and putative nuclear localization signals. hPRP4 phosphorylated and interacted with SF2/ASF, one of the essential splicing factors. Indirect immunofluorescence analysis revealed that endogenous hPRP4 was distributed in a nuclear speckled pattern and colocalized with SF2/ASF in HeLa S3 cells. Furthermore, hPRP4 interacted directly with Clk1 on its COOH terminus, and the arginine/serine-rich domain of hPRP4 was phosphorylated by Clk1 in vitro. Overexpression of Clk1 caused redistribution of hPRP4, from the speckled to the diffuse pattern in nucleoplasm, whereas inactive mutant of Clk1 caused no change of hPRP4 localization. These findings suggest that the NH(2)-terminal region of hPRP4 may play regulatory roles under an unidentified signal transduction pathway through Clk1. PMID- 11418605 TI - Role of individual disulfide bonds in the structural maturation of a low molecular weight glutenin subunit. AB - Gliadins and glutenins are the major storage proteins that accumulate in wheat endosperm cells during seed development. Although gliadins are mainly monomeric, glutenins consist of very large disulfide-linked polymers made up of high molecular weight and low molecular weight subunits. These polymers are among the largest protein molecules known in nature and are the most important determinants of the viscoelastic properties of gluten. As a first step toward the elucidation of the folding and assembly pathways that lead to glutenin polymer formation, we have exploited an in vitro system composed of wheat germ extract and bean microsomes to examine the role of disulfide bonds in the structural maturation of a low molecular weight glutenin subunit. When conditions allowing the formation of disulfide bonds were established, the in vitro synthesized low molecular weight glutenin subunit was recovered in monomeric form containing intrachain disulfide bonds. Conversely, synthesis under conditions that did not favor the formation of disulfide bonds led to the production of large aggregates from which the polypeptides could not be rescued by the post-translational generation of a more oxidizing environment. These results indicate that disulfide bond formation is essential for the conformational maturation of the low molecular weight glutenin subunit and suggest that early folding steps may play an important role in this process, allowing the timely pairing of critical cysteine residues. To determine which cysteines were important to maintain the protein in monomeric form, we prepared a set of mutants containing selected cysteine to serine substitutions. Our results show that two conserved cysteine residues form a critical disulfide bond that is essential in preventing the exposure of adhesive domains and the consequent formation of aberrant aggregates. PMID- 11418606 TI - Acute phase protein alpha 1-acid glycoprotein interacts with plasminogen activator inhibitor type 1 and stabilizes its inhibitory activity. AB - alpha(1)-Acid glycoprotein, one of the major acute phase proteins, was found to interact with plasminogen activator inhibitor type 1 (PAI-1) and to stabilize its inhibitory activity toward plasminogen activators. This conclusion is based on the following observations: (a) alpha(1)-acid glycoprotein was identified to bind PAI-1 by a yeast two-hybrid system. Three of 10 positive clones identified by this method to interact with PAI-1 contained almost the entire sequence of alpha(1)-acid glycoprotein; (b) this protein formed complexes with PAI-1 that could be immunoprecipitated from both the incubation mixtures and blood plasma by specific antibodies to either PAI-1 or alpha(1)-acid glycoprotein. Such complexes could be also detected by a solid phase binding assay; and (c) the real-time bimolecular interactions monitored by surface plasmon resonance indicated that the complex of alpha(1)-acid glycoprotein with PAI-1 is less stable than that formed by vitronectin with PAI-1, but in both cases, the apparent K(D) values were in the range of strong interactions (4.51 + 1.33 and 0.58 + 0.07 nm, respectively). The on rate for binding of PAI-1 to alpha(1)-glycoprotein or vitronectin differed by 2-fold, indicating much faster complex formation by vitronectin than by alpha(1)-acid glycoprotein. On the other hand, dissociation of PAI-1 bound to vitronectin was much slower than that from the alpha(1)-acid glycoprotein, as indicated by 4-fold lower k(off) values. Furthermore, the PAI-1 activity toward urokinase-type plasminogen activator and tissue-type plasminogen activator was significantly prolonged in the presence of alpha(1)-acid glycoprotein. These observations suggest that the complex of PAI-1 with alpha(1) acid glycoprotein can play a role as an alternative reservoir of the physiologically active form of the inhibitor, particularly during inflammation or other acute phase reactions. PMID- 11418607 TI - Characterization of human mucin gene MUC4 promoter: importance of growth factors and proinflammatory cytokines for its regulation in pancreatic cancer cells. AB - The human mucin gene MUC4 encodes a large transmembrane mucin that is thought to play important roles in tumor cell biology and that is overexpressed in human pancreatic carcinomas. In this report, we describe the structure and functional activity of the 5'-flanking region, including 1.0 kilobase of the promoter. The long 5'-untranslated region (2.7 kilobases) is characterized by a high content of GC in its 3'-end. The first TATA box was located at -2672/-2668. Multiple transcription start sites and a high density of putative binding sites for Sp1 (GC and CACCC boxes), AP-1/-2/-4, cAMP-responsive element-binding protein, GATA, GR, and STAT transcription factors were found within the 5'-flanking region. Transcriptional activity of the promoter was assessed using pGL3-luciferase deletion mutants in two MUC4-expressing (CAPAN-1 and CAPAN-2) and one nonexpressing (PANC-1) pancreatic cancer cell line. Two highly active fragments ( 219/-1 and -2781/-2572) that drive MUC4 transcription in CAPAN-1 and CAPAN-2 cells were identified. Gel retardation assays indicated that Sp1 and Sp3 bind to cognate cis-elements found in the 5'-flanking region and that Sp1 transactivates, whereas Sp3 inhibits the GC-rich region (-464/-1) in CAPAN-2 cells. Activation of protein kinase C with phorbol ester and treatment of cells with epidermal growth factor and transforming growth factor-alpha resulted in up-regulation of the promoter. Tumor necrosis factor-alpha and interferon (IFN)-gamma inflammatory cytokines had no or mild effect on MUC4 transcriptional activity when used alone. However, a very strong synergistic effect (10-12-fold activation) between IFN gamma and tumor necrosis factor-alpha or IFN-gamma and transforming growth factor alpha was obtained in CAPAN-2 cells. Altogether these results demonstrate that the 5'-flanking region of MUC4 contains epithelial cell-specific, positive, and negative regulatory cis-elements, that Sp1/Sp3 are important regulators of MUC4 basal expression, and that its regulation in pancreatic cancer cells involves complex interplay between several signaling pathways. PMID- 11418608 TI - The mood stabilizer valproic acid activates mitogen-activated protein kinases and promotes neurite growth. AB - The mood-stabilizing agents lithium and valproic acid (VPA) increase DNA binding activity and transactivation activity of AP-1 transcription factors, as well as the expression of genes regulated by AP-1, in cultured cells and brain regions involved in mood regulation. In the present study, we found that VPA activated extracellular signal-regulated kinase (ERK), a kinase known to regulate AP-1 function and utilized by neurotrophins to mediate their diverse effects, including neuronal differentiation, neuronal survival, long term neuroplasticity, and potentially learning and memory. VPA-induced activation of ERK was blocked by the mitogen-activated protein kinase/ERK kinase inhibitor PD098059 and dominant negative Ras and Raf mutants but not by dominant-negative stress-activated protein kinase/ERK kinase and mitogen-activated protein kinase kinase 6 mutants. VPA also increased the expression of genes regulated by the ERK pathway, including growth cone-associated protein 43 and Bcl-2, promoted neurite growth and cell survival, and enhanced norepinephrine uptake and release. These data demonstrate that VPA is an ERK pathway activator and produces neurotrophic effects. PMID- 11418609 TI - Two basic regions of NCp7 are sufficient for conformational conversion of HIV-1 dimerization initiation site from kissing-loop dimer to extended-duplex dimer. AB - Nucleocapsid (NC) protein possesses nucleotide-annealing activities, which are used in various processes in retroviral life cycle. As conserved characters, the NC proteins have one or two zinc fingers of CX(2)CX(4)HX(4)C motif surrounded by basic amino acid sequences. Requirement of the zinc fingers for the annealing activities of NC protein remains controversial. In this study, we focused the requirement in the process of maturation of dimeric viral RNA. Discrimination between immature and mature dimers of synthetic RNA corresponding to the dimerization initiation site of human immunodeficiency virus type 1 (HIV-1) genomic RNA was performed based on their Mg(2+)-dependent stability in gel electrophoreses and on their distinct signal pattern from NMR analysis of imino protons. Chaperoning activity of the HIV-1 NC protein, NCp7, and its fragments for maturation of dimeric RNA was investigated using these experimental systems. We found that the two basic regions flanking the N-terminal zinc finger of NCp7, which are connected by two glycine residues instead of the zinc finger, were sufficient, although about 10 times the amounts of peptide were needed in comparison with intact NCp7. Further, it was found that the amount of basic residues rather than the amino acid sequence itself is important for the activity. The zinc fingers may involve the binding affinity and/or such a possible specific binding of NCp7 to dimerization initiation site dimer that leads to the maturation reaction. PMID- 11418610 TI - Redundant exonuclease involvement in Escherichia coli methyl-directed mismatch repair. AB - Previous biochemical analysis of Escherichia coli methyl-directed mismatch repair implicates three redundant single-strand DNA-specific exonucleases (RecJ, ExoI, and ExoVII) and at least one additional unknown exonuclease in the excision reaction (Cooper, D. L., Lahue, R. S., and Modrich, P. (1993) J. Biol. Chem. 268, 11823-11829). We show here that ExoX also participates in methyl-directed mismatch repair. Analysis of the reaction with crude extracts and purified components demonstrated that ExoX can mediate repair directed from a strand signal 3' of a mismatch. Whereas extracts of all possible single, double, and triple exonuclease mutants displayed significant residual mismatch repair, extracts deficient in RecJ, ExoI, ExoVII, and ExoX exonucleases were devoid of normal repair activity. The RecJ(-) ExoVII(-) ExoI(-) ExoX(-) strain displayed a 7-fold increase in mutation rate, a significant increase, but less than that observed for other blocks of the mismatch repair pathway. This elevation is epistatic to deficiency for MutS, suggesting an effect via the mismatch repair pathway. Our other work (Burdett, V., Baitinger, C., Viswanathan, M., Lovett, S. T., and Modrich, P. (2001) Proc. Natl. Acad. Sci. U. S. A. 98, 6765-6770) suggests that mutants are under-recovered in the exonuclease-deficient strain due to loss of viability that is triggered by mismatched base pairs in this genetic background. The availability of any one exonuclease is enough to support full mismatch correction, as evident from the normal mutation rates of all triple mutants. Because three of these exonucleases possess a strict polarity of digestion, this suggests that mismatch repair can occur exclusively from a 3' or a 5' direction to the mismatch, if necessary. PMID- 11418611 TI - RGS2 promotes adipocyte differentiation in the presence of ligand for peroxisome proliferator-activated receptor gamma. AB - The events at the earliest stage of adipocyte differentiation are yet to be fully elucidated. Previously, we cloned the genes that are induced at the beginning of the differentiation of mouse 3T3-L1 preadipocyte cells. We found that the gene expression of regulators of G protein signaling-2 (RGS2) rapidly increased after the addition of inducers and decreased at 3-12 h. The expression pattern of RGS2 mRNAs differed among growth-arrested and proliferating 3T3-L1 cells and NIH-3T3 cells, indicating a specificity for adipogenesis. Here we report that the ectopic expression of RGS2 using a retroviral system in mouse NIH-3T3 cells promotes adipogenesis only in the presence of BRL49653, which is a ligand for the peroxisome proliferator-activated receptor gamma (PPARgamma). These results strongly suggest that RGS2 play a crucial role in the program of adipocyte differentiation and may contribute to the function of PPARgamma. PMID- 11418612 TI - T cell activation induces direct binding of the Crk adapter protein to the regulatory subunit of phosphatidylinositol 3-kinase (p85) via a complex mechanism involving the Cbl protein. AB - The Crk adapter proteins are assumed to play a role in T lymphocyte activation because of their induced association with tyrosine-phosphorylated proteins, such as ZAP-70 and Cbl, and with the phosphatidylinositol 3kinase regulatory subunit, p85, following engagement of the T cell antigen receptor. Although the exact mechanism of interaction between these molecules has not been fully defined, it has been generally accepted that Crk, ZAP-70, and p85 interact with tyrosine phosphorylated Cbl, which serves as a major scaffold protein in activated T lymphocytes. Our present results demonstrate a cell activation-dependent reciprocal co-immunoprecipitation of CrkII and p85 from lysates of Jurkat T cells and a direct binding of CrkII to p85 in an overlay assay. The use of bead immobilized GST fusion proteins indicated a complex mechanism of interaction between CrkII and p85 involving two distinct and mutually independent regions in each molecule. A relatively high affinity binding of the CrkII-SH3(N) domain to p85 and the p85-proline-B cell receptor-proline (PBP) region to CrkII was observed in lysates of either resting or activated T cells. Direct physical interaction between the CrkII-SH3(N) and the p85-PBP domain was demonstrated using recombinant fusion proteins and was further substantiated by binding competition studies. In addition, immobilized fusion proteins possessing the CrkII-SH2 and p85-SH3 domains were found to pull down p85 and CrkII, respectively, but only from lysates of activated T cells. Nevertheless, the GST CrkII-SH2 fusion protein was unable to mediate direct association with p85 from lysates of either resting or activated T cells. Our results support a model in which T cell activation dependent conformational changes in CrkII and/or p85 promote an initial direct or indirect low affinity interaction between the two molecules, which is then stabilized by a secondary high affinity interaction mediated by direct binding of the CrkII-SH3(N) to the p85-PBP domain. PMID- 11418613 TI - Shigella protein IpaH(9.8) is secreted from bacteria within mammalian cells and transported to the nucleus. AB - Various pathogenic bacteria such as Shigella deliver effector proteins into mammalian cells via the type III secretion system. The delivered Shigella effectors have been shown to variously affect host functions required for efficient bacterial internalization into the cells. In the present study, we investigated the IpaH proteins for their ability to be secreted via the type III secretion system and their fate in mammalian cells. Upon incubation in a medium containing Congo red, the bacteria secrete IpaH into the medium, but secretion of IpaH occurs later than that of IpaBCD. Immunofluorescence microscopy indicated that IpaH(9.8) is secreted from intracellular bacteria and transported into the nucleus. On microinjection of the protein, intracellular IpaH(9.8) is accumulated at one place around the nucleus and transported into the nucleus. This movement seems to be dependent on the microtubule network, since nuclear accumulation of IpaH(9.8) is inhibited in cells treated with microtubule-destabilizing agents. In nuclear import assay, IpaH(9.8) was efficiently transported into the nucleus, which was completely blocked by treatment with wheat germ agglutinin. The nuclear transport of IpaH(9.8) does not depend on host cytosolic factors but is partially dependent on ATP/GTP, suggesting that, like beta-catenin, IpaH(9.8) secreted from intracellular Shigella can be transported into the nucleus. PMID- 11418614 TI - Specific residues within the alpha 2 integrin subunit cytoplasmic domain regulate migration and cell cycle progression via distinct MAPK pathways. AB - The alpha(2) integrin subunit cytoplasmic domain is necessary for epidermal growth factor (EGF)-stimulated chemotactic migration and insulin-dependent entry into S-phase of mammary epithelial cells adherent to type I collagen. Truncation mutants revealed that the seven amino acids, KYEKMTK, in addition to the GFFKR motif were sufficient for these functions. Mutation of tyrosine 1134 to alanine inhibited the ability of the cells to phosphorylate p38 MAPK and to migrate in response to EGF but had only a modest effect on the ability of the cells to induce sustained phosphorylation of the ERK MAPK, to up-regulate cyclin E and cdk2 expression, and to enter S-phase when adherent to type I collagen. Conversely, mutation of the lysine 1136 inhibited the ability of the cells to increase cyclin E and cdk2 expression, to maintain long term phosphorylation of the ERK MAPK, and to enter S-phase but had no effect on the ability of the cells to phosphorylate the p38 MAPK or to migrate on type I collagen in response to EGF. Methionine 1137 was essential for both migration and entry into S-phase. Thus, distinctly different structural elements of the alpha(2) integrin cytoplasmic domain are required to engage the signaling pathways leading to cell migration or cell cycle progression. PMID- 11418615 TI - Negative regulation of human fibrinogen gene expression by peroxisome proliferator-activated receptor alpha agonists via inhibition of CCAAT box/enhancer-binding protein beta. AB - Fibrinogen is a coagulation factor and an acute phase reactant up-regulated by inflammatory cytokines, such as interleukin 6 (IL-6). Elevated plasma fibrinogen levels are associated with coronary heart diseases. Fibrates are clinically used hypolipidemic drugs that act via the nuclear receptor peroxisome proliferator activated receptor alpha (PPAR alpha). In addition, most fibrates also reduce plasma fibrinogen levels, but the molecular mechanism is unknown. In this study, we demonstrate that fibrates decrease basal and IL-6-stimulated expression of the human fibrinogen-beta gene in human primary hepatocytes and hepatoma HepG2 cells. Fibrates diminish basal and IL-6-induced fibrinogen-beta promoter activity, and this effect is enhanced in the presence of co-transfected PPAR alpha. Site directed mutagenesis experiments demonstrate that PPAR alpha activators decrease human fibrinogen-beta promoter activity via the CCAAT box/enhancer-binding protein (C/EBP) response element. Co-transfection of the transcriptional intermediary factor glucocorticoid receptor-interacting protein 1/transcriptional intermediary factor 2 (GRIP1/TIF2) enhances fibrinogen-beta gene transcription and alleviates the repressive effect of PPAR alpha. Co-immunoprecipitation experiments demonstrate that PPAR alpha and GRIP1/TIF2 physically interact in vivo in human liver. These data demonstrate that PPAR alpha agonists repress human fibrinogen gene expression by interference with the C/EBP beta pathway through titration of the coactivator GRIP1/TIF2. We observed that the anti inflammatory action of PPAR alpha is not restricted to fibrinogen but also applies to other acute phase genes containing a C/EBP response element; it also occurs under conditions in which the stimulating action of IL-6 is potentiated by dexamethasone. These findings identify a novel molecular mechanism of negative gene regulation by PPAR alpha and reveal the direct implication of PPAR alpha in the modulation of the inflammatory gene response in the liver. PMID- 11418616 TI - Synemin may function to directly link muscle cell intermediate filaments to both myofibrillar Z-lines and costameres. AB - Synemin is a large intermediate filament (IF) protein that has been identified in all types of muscle cells in association with desmin- and/or vimentin-containing IFs. Our previous studies (Bellin, R. M., Sernett, S. W., Becker, B., Ip, W., Huiatt, T. W., and Robson, R. M. (1999) J. Biol. Chem. 274, 29493-29499) demonstrated that synemin forms heteropolymeric IFs with major IF proteins and contains a binding site for the myofibrillar Z-line protein alpha-actinin. By utilizing blot overlay assays, we show herein that synemin also interacts with the costameric protein vinculin. Furthermore, extensive assays utilizing the Gal4 yeast two-hybrid system demonstrate interactions of synemin with desmin and vimentin and additionally define more precisely the protein subdomains involved in the synemin/alpha-actinin and synemin/vinculin interactions. The C-terminal approximately 300-amino acid region of synemin binds to the N-terminal head and central rod domains of alpha-actinin and the approximately 150-amino acid C terminal tail of vinculin. Overall, these interactions indicate that synemin may anchor IFs to myofibrillar Z-lines via interactions with alpha-actinin and to costameres at the sarcolemma via interactions with vinculin and/or alpha-actinin. These linkages would enable the IFs to directly link all cellular myofibrils and to anchor the peripheral layer of myofibrils to the costameres. PMID- 11418617 TI - Arrestin specificity for G protein-coupled receptors in human airway smooth muscle. AB - Despite a widely accepted role of arrestins as "uncouplers" of G protein-coupled receptor (GPCR) signaling, few studies have demonstrated the ability of arrestins to affect second messenger generation by endogenously expressed receptors in intact cells. In this study we demonstrate arrestin specificity for endogenous GPCRs in primary cultures of human airway smooth muscle (HASM). Expression of arrestin-green fluorescent protein (ARR2-GFP or ARR3-GFP) chimeras in HASM significantly attenuated isoproterenol (beta(2)-adrenergic receptor (beta(2)AR) mediated)- and 5'-(N-ethylcarboxamido)adenosine (A2b adenosine receptor-mediated) stimulated cAMP production, with fluorescent microscopy demonstrating agonist promoted redistribution of cellular ARR2-GFP into a punctate formation. Conversely, prostaglandin E(2) (PGE(2))-mediated cAMP production was unaffected by arrestin-GFP, and PGE(2) had little effect on arrestin-GFP distribution. The pharmacological profile of various selective EP receptor ligands suggested a predominantly EP2 receptor population in HASM. Further analysis in COS-1 cells revealed that ARR2-GFP expression increased agonist-promoted internalization of wild type beta(2)AR and EP4 receptors, whereas EP2 receptors remained resistant to internalization. However, expression of an arrestin whose binding to GPCRs is largely independent of receptor phosphorylation (ARR2(R169E)-GFP) enabled substantial agonist-promoted EP2 receptor internalization, increased beta(2)AR internalization to a greater extent than did ARR2-GFP, yet promoted EP4 receptor internalization to the same degree as did ARR2-GFP. Signaling via endogenous EP4 receptors in CHO-K1 cells was attenuated by ARR2-GFP expression, whereas ARR2(R169E)-GFP expression in HASM inhibited EP2 receptor-mediated cAMP production. These findings demonstrate differential effects of arrestins in altering endogenous GPCR signaling in a physiologically relevant cell type and reveal a variable dependence on receptor phosphorylation in dictating arrestin receptor interaction. PMID- 11418618 TI - Conformational intermediate of the amyloidogenic protein beta 2-microglobulin at neutral pH. AB - Aggregation and fibrillation of beta(2)-microglobulin are hallmarks of dialysis related amyloidosis. We characterize perturbations of the native conformation of beta(2)-microglobulin that may precede fibril formation. For a beta(2) microglobulin variant cleaved at lysine 58, we show using capillary electrophoresis that two conformers spontaneously exist in aqueous buffers at neutral pH. Upon treatment of wild-type beta(2)-microglobulin with acetonitrile or trifluoroethanol, two conformations were also observed. These conformations were in equilibrium dependent on the sample temperature and the percentage of organic solvent present. Circular dichroism showed a loss of beta-structures and gain of alpha-helices. Reversal to the native conformation occurred when removing the organics. Affinity capillary electrophoresis experiments showed increased specific interactions of the nonnative beta(2)-microglobulin conformation with the dyes 8-anilino-1-naphthalene sulfonic acid and Congo red. The observations may relate to early folding events prior to amyloid fibrillation and facilitate the development of methods to detect and inhibit pro-amyloid protein and peptide conformations. PMID- 11418619 TI - Human glutathione transferase A3-3, a highly efficient catalyst of double-bond isomerization in the biosynthetic pathway of steroid hormones. AB - The cDNA of a novel human glutathione transferase (GST) of the Alpha class was cloned, and the corresponding protein, denoted GST A3-3, was heterologously expressed and characterized. GST A3-3 was found to efficiently catalyze obligatory double-bond isomerizations of Delta(5)-androstene-3,17-dione and Delta(5)-pregnene-3,20-dione, precursors to testosterone and progesterone, respectively, in steroid hormone biosynthesis. The catalytic efficiency (k(cat)/K(m)) with Delta(5)-androstene-3,17-dione was determined as 5 x 10(6) m( 1) s(-1), which is considerably higher than with any other GST substrate tested. The rate of acceleration afforded by GST A3-3 is 6 x 10(8) based on the ratio between k(cat) and the rate constant for the nonenzymatic isomerization of Delta(5)-androstene-3,17-dione. Besides being high in absolute numbers, the k(cat)/K(m) value of GST A3-3 exceeds by a factor of approximately 230 that of 3beta-hydroxysteroid dehydrogenase/isomerase, the enzyme generally considered to catalyze the Delta(5)-Delta(4) double-bond isomerization. Furthermore, GSTA3 specific polymerase chain reaction analysis of cDNA libraries from various tissues showed a message only in those characterized by active steroid hormone biosynthesis, indicating a selective expression of GST A3-3 in these tissues. Based on this finding and the high activity with steroid substrates, we propose that GST A3-3 has evolved to catalyze isomerization reactions that contribute to the biosynthesis of steroid hormones. PMID- 11418620 TI - Extracellular fibrinogen-binding protein, Efb, from Staphylococcus aureus blocks platelet aggregation due to its binding to the alpha-chain. AB - Extracellular fibrinogen-binding protein (Efb) secreted by Staphylococcus aureus has previously been shown to contribute to pathogenesis in a rat wound infection model. Also antibodies against Efb exhibited a protective effect in a mouse mastitis model. The interaction between Efb and fibrinogen is divalent, with one binding site within the N-terminal repeat region in Efb and one at the C terminus. In this study we show that the distal D domain of fibrinogen contains at least one of the binding domains recognized by Efb. Efb stimulates fibrinogen binding to ADP-activated platelets. Furthermore, Efb inhibits ADP-induced, fibrinogen-dependent platelet aggregation in a concentration-dependent manner. This implies that Efb modifies platelet function by amplifying a non-functional interaction between fibrinogen and platelets. Efb recognizes the A alpha-chain of the D fragment of fibrinogen. The RGD sequence on the A alpha-chain is located close to the region recognized by Efb and contains a putative binding site for the platelet integrin GPIIb/IIIa receptor complex involved in platelet aggregation. PMID- 11418621 TI - Characterization of a dehydrogenase activity responsible for oxidation of 11-cis retinol in the retinal pigment epithelium of mice with a disrupted RDH5 gene. A model for the human hereditary disease fundus albipunctatus. AB - In the vertebrate retina, the final step of visual chromophore production is the oxidation of 11-cis-retinol to 11-cis-retinal. This reaction is catalyzed by 11 cis-retinol dehydrogenases (11-cis-RDHs), prior to the chromophore rejoining with the visual pigment apo-proteins. The RDH5 gene encodes a dehydrogenase that is responsible for the majority of RDH activity. In humans, mutations in this gene are associated with fundus albipunctatus, a disease expressed by delayed dark adaptation of both cones and rods. In this report, an animal model for this disease, 11-cis-rdh-/- mice, was used to investigate the flow of retinoids after a bleach, and microsomal membranes from the retinal pigment epithelium of these mice were employed to characterize remaining enzymatic activities oxidizing 11 cis-retinol. Lack of 11-cis-RDH leads to an accumulation of cis-retinoids, particularly 13-cis-isomers. The analysis of 11-cis-rdh-/- mice showed that the RDH(s) responsible for the production of 11-cis-retinal displays NADP-dependent specificity toward 9-cis- and 11-cis-retinal but not 13-cis-retinal. The lack of 13-cis-RDH activity could be a reason why 13-cis-isomers accumulate in the retinal pigment epithelium of 11-cis-rdh-/- mice. Furthermore, our results provide detailed characterization of a mouse model for the human disease fundus albipunctatus and emphasize the importance of 11-cis-RDH in keeping the balance between different components of the retinoid cycle. PMID- 11418622 TI - Phosphatidylinositol 3-kinase is necessary but not sufficient for thrombopoietin induced proliferation in engineered Mpl-bearing cell lines as well as in primary megakaryocytic progenitors. AB - Thrombopoietin and its receptor (Mpl) support survival and proliferation in megakaryocyte progenitors and in BaF3 cells engineered to stably express Mpl (BaF3/Mpl). The binding of thrombopoietin to Mpl activates multiple kinase pathways, including the Jak/STAT, Ras/Raf/MAPK, and phosphatidylinositol 3-kinase pathways, but it is not clear how these kinases promote cell cycling. Here, we show that thrombopoietin induces phosphatidylinositol 3-kinase and that phosphatidylinositol 3-kinase is required for thrombopoietin-induced cell cycling in BaF3/Mpl cells and in primary megakaryocyte progenitors. Treatment of BaF3/Mpl cells and megakaryocytes with the phosphatidylinositol 3-kinase inhibitor LY294002 inhibited mitotic and endomitotic cell cycl-ing. BaF3/Mpl cells treated with thrombopoietin and LY294002 were blocked in G(1), whereas megakaryocyte progenitors treated with thrombopoietin and LY294002 showed both a G(1) and a G(2) cell cycle block. Expression of constitutively active Akt in BaF3/Mpl cells restored the ability of thrombopoietin to promote cell cycling in the presence of LY294002. Constitutively active Akt was not sufficient to drive proliferation of BaF3/Mpl cells in the absence of thrombopoietin. We conclude that in BaF3/Mpl cells and megakaryocyte progenitors, thrombopoietin-induced phosphatidylinositol 3-kinase activity is necessary but not sufficient for thrombopoietin-induced cell cycle progression. Phosphatidylinositol 3-kinase activity is likely to be involved in regulating the G(1)/S transition. PMID- 11418623 TI - Cutting edge: the common gamma-chain is an indispensable subunit of the IL-21 receptor complex. AB - The common gamma-chain (gamma(c)) is an indispensable subunit of the functional receptor complexes for IL-4, IL-7, IL-9, and IL-15 as well as IL-2. Here we show that the gamma(c) is also shared with the IL-21R complex. Although IL-21 binds to the IL-21R expressed on gamma(c)-deficient ED40515(-) cells, IL-21 is unable to transduce any intracytoplasmic signals. However, in EDgamma-16 cells, a gamma(c) transfected ED40515(-) cell line, IL-21 binds to the IL-21R and can activate Janus kinase (JAK)1, JAK3, STAT1, and STAT3. The chemical cross-linking study reveals the direct binding of IL-21 to the gamma(c). These data clearly demonstrate that the gamma(c) is an indispensable subunit of the functional IL 21R complex. PMID- 11418624 TI - Cutting edge: Ly49A inhibits TCR/CD3-induced apoptosis and IL-2 secretion. AB - To evaluate the importance of Ly49A on TCR-induced cellular events, we established clones of the 1F2 T cell hybridoma expressing either Ly49A or a chimeric version, Ly49A/H, where the Ly49A cytoplasmic domain has been replaced by the Ly49H cytoplasmic domain. Ligation of Ly49A, but not Ly49A/H, with its ligand H-2D(d) or anti-Ly49A mAbs caused a specific inhibition of TCR/CD3-induced IL-2 secretion. Moreover, flow cytometry analysis of hypodiploid DNA and annexin V binding revealed that ligation of Ly49A protected cells from apoptosis induced by anti-CD3 mAbs or Ag. In contrast, ligation of the Ly49A/H chimeric receptor had no antiapoptotic effect. In addition, engagement of Ly49A selectively inhibited TCR-induced Fas ligand expression whereas TCR-induced Fas expression was not significantly affected. Expression of Ly49 inhibitory receptors on T cells may represent an important mechanism for the regulation of T cell survival in vivo by inhibiting TCR-induced apoptosis and IL-2 secretion. PMID- 11418625 TI - Cutting edge: antibody-mediated cessation of hemotropic infection by the intraerythrocytic mouse pathogen Bartonella grahamii. AB - The genus Bartonella includes important human-specific and zoonotic pathogens which cause intraerythrocytic bacteremia in their mammalian reservoir host(s). It is accepted that cellular immunity plays a decisive role in the host's defense against most intracellular bacteria. Bartonella sp. infection in the immunocompetent host typically leads to immunity against homologous challenge. The basis of this immunity, be it cellular or humoral, is unclear. In this study, the course of Bartonella grahamii bacteremia in immunocompetent and immunocompromised mice was compared. In immunocompetent hosts, the bacteremia is transient and induces a strong humoral immune response. In contrast, bacteremia persists in immunocompromised B and T cell-deficient mice. Immune serum transfer beginning with day 6 postinfection to B cell-deficient mice unable to produce Igs converted the persistent bacteremia to a transient course indistinguishable from that of immunocompetent animals. These data demonstrate an essential role for specific Abs in abrogating the intraerythrocytic bacteremia of B. grahamii in mice. PMID- 11418626 TI - Cutting edge: germinal centers can be induced in the absence of T cells. AB - Immunization of mice containing mutations that inactivate the TCR Cbeta and Cdelta genes with the T cell-independent (TI) type 2 Ag (4-hydroxy-3 nitrophenyl)acetyl-Ficoll induces clusters of peanut agglutinin-binding B cells in the spleen. These clusters are histologically indistinguishable from germinal centers (GCs) typical of T cell-dependent immune responses. They are located in follicles, and contain mature follicular dendritic cells, immune complex deposits, and B cells that display the phenotypic qualities of conventional GC B cells. However, the kinetics of this TI GC response differ from T cell-dependent GC responses in being rapidly induced and of short duration. Moreover, the Ab V genes expressed in TI GCs have not undergone somatic hypermutation. Therefore, T cells may be required for B cell differentiation processes associated with the intermediate and latter stages of the GC reaction, but they are dispensable for the induction and initial development of this response. PMID- 11418627 TI - Molecular characterization of B cell clonal expansions in the liver of chronically hepatitis C virus-infected patients. AB - PCR DNA amplification of IgH genes was performed on liver biopsy samples of 42 unselected hepatitis C virus (HCV)-positive patients. Genotypic analysis and signal amplification by branched DNA were used to characterize and quantitate HCV RNA genomic sequences. Intraportal lymphoid follicle-like structures were isolated from surrounding hepatocytes by microdissection technique. IgH VDJ PCR products were cloned and sequenced. IgH VDJ gene rearrangements were detected in the liver of 26 (62%) patients. Unequivocal monoclonal or oligoclonal patterns of B cell expansions were found in 14 (33.3%) and 12 (28.6%) patients, respectively. Patients with intrahepatic B cell monoclonal expansions showed liver HCV RNA levels higher than those with oligoclonal or polyclonal features (1106.4 +/- 593.5 vs 677.3 +/- 424.3 vs 406.2 +/- 354.3 pg HCV RNA/g tissue; p = 0.048 and p = 0.001, respectively). Although a single dominant band was obtained with total DNA, characterization of DNA recovered from intraportal inflammatory aggregates resulted in the detection of multiple IgH VDJ gene rearrangements, pointing to an oligoclonal pattern of lymphoproliferation. Cloning and sequence analyses showed that B cell clonalities were differently distributed in adjacent portal tracts of the same liver area. In addition, HCV RNA genomic sequences could be consistently amplified from each of the portal inflammatory aggregates examined. These data support the concept that in chronic HCV infection the intrahepatic B cell repertoire is frequently clonally restricted and that HCV may have a direct role in sustaining in situ B cell proliferation. PMID- 11418628 TI - 11 Beta-hydroxysteroid dehydrogenase type 1 is induced in human monocytes upon differentiation to macrophages. AB - 11beta-hydroxysteroid dehydrogenases (11beta-HSD) perform prereceptor metabolism of glucocorticoids through interconversion of the active glucocorticoid, cortisol, with inactive cortisone. Although the immunosuppressive and anti inflammatory activities of glucocorticoids are well documented, the expression of 11beta-HSD enzymes in immune cells is not well understood. Here we demonstrate that 11beta-HSD1, which converts cortisone to cortisol, is expressed only upon differentiation of human monocytes to macrophages. 11beta-HSD1 expression is concomitant with the emergence of peroxisome proliferator activating receptor gamma, which was used as a surrogate marker of monocyte differentiation. The type 2 enzyme, 11beta-HSD2, which converts cortisol to cortisone, was not detectable in either monocytes or cultured macrophages. Incubation of monocytes with IL-4 or IL-13 induced 11beta-HSD1 activity by up to 10-fold. IFN-gamma, a known functional antagonist of IL-4 and IL-13, suppressed the induction of 11beta-HSD1 by these cytokines. THP-1 cells, a human macrophage-like cell line, expressed 11beta-HSD1 and low levels of 11beta-HSD2. The expression of 11beta-HSD1 in these cells is up-regulated 4-fold by LPS. In summary, we have shown strong expression of 11beta-HSD1 in cultured human macrophages and THP-1 cells. The presence of the enzyme in these cells suggests that it may play a role in regulating the immune function of these cells. PMID- 11418629 TI - Structural features of nonpeptide prenyl pyrophosphates that determine their antigenicity for human gamma delta T cells. AB - Human Vgamma2Vdelta2(+) T cells proliferate in vivo during many microbial infections. We have found that Vgamma2Vdelta2(+) T cells recognize nonpeptide prenyl pyrophosphates and alkylamines. We now have defined structural features that determine the antigenicity of prenyl pyrophosphates by testing synthetic analogs for bioactivity. We find that the carbon chain closest to the pyrophosphate moiety plays the major role in determining bioactivity. Changes in this area, such as the loss of a double bond, abrogated bioactivity. The loss of a phosphate from the pyrophosphate moiety also decreased antigenicity 100- to 200 fold. However, nucleotide monophosphates could be added with minimal changes in bioactivity. Longer prenyl pyrophosphates also retained bioactivity. Despite differences in CDR3 sequence, Vgamma2Vdelta2(+) clones and a transfectant responded similarly. Ag docking into a Vgamma2Vdelta2 TCR model reveals a potential binding site in germline regions of the Vgamma2Jgamma1.2 CDR3 and Vdelta2 CDR2 loops. Thus, Vgamma2Vdelta2(+) T cells recognize a core carbon chain and pyrophosphate moiety. This recognition is relatively unaffected by additions at distal positions to the core Ag unit. PMID- 11418630 TI - Expression of active protein kinase B in T cells perturbs both T and B cell homeostasis and promotes inflammation. AB - The molecular mechanisms that contribute to autoimmunity remain poorly defined. While inflammation is considered to be one of the major checkpoints in autoimmune disease progression, very little is known about the initiating events that trigger inflammation. We have studied transgenic mice expressing the prosurvival molecule protein kinase B/Akt under control of a T cell-specific CD2 promoter. In this study, we demonstrate that aged mice develop lymphadenopathy and splenomegaly that result from an accumulation of CD4, CD8, and unexpectedly B cells. An increased proportion of T cells express activation markers, while T cell proliferative responses remain normal. B cells are hyperproliferative in response to anti-IgM F(ab')(2) and anti-CD40, and increased IgA and IgG2a were found in the sera. In addition, a profound multiorgan lymphocytic infiltration is observed, and T cells from these mice display a defect in Fas-mediated apoptosis, which may be the mechanism underlying this phenotype. Therefore, T cell expression of active protein kinase B can alter T cell homeostasis, indirectly influence B cell homeostasis, and promote inflammation in vivo. PMID- 11418631 TI - The distinct roles of T cell-derived cytokines and a novel follicular dendritic cell-signaling molecule 8D6 in germinal center-B cell differentiation. AB - Germinal center-B (GC-B) cells differentiate into memory B cells and plasma cells (PC) through interaction with T cells and follicular dendritic cells (FDC). Activated T cell and FDC play distinct roles in this process. The detailed kinetic experiments revealed that cytokines secreted by activated T cells determined the pathway of GC-B cell differentiation. IL-4 directs GC-B cells to differentiate into memory B cells, whereas IL-10 steers them into PC. FDC/HK cells do not direct either pathway, but provide signals for proliferation of GC-B cells. A novel FDC-signaling molecule 8D6 (FDC-SM-8D6) produced by FDC augments PC generation in the GC. FDC-SM-8D6-specific mAb blocked PC generation and IgG secretion but not memory B cell proliferation. COS cells expressing FDC-SM-8D6 enhanced GC-B cell proliferation and Ab secretion, which was blocked by mAb 8D6. In the cultures with B cell subsets, PC generation was inhibited by mAb 8D6 in the cultures with CD27(+) B cells but not in the culture with CD27(-) B cells, suggesting that CD27(+) PC precursor is the specific target of FDC-SM-8D6 stimulation. PMID- 11418632 TI - Lymphotactin expression by engineered myeloma cells drives tumor regression: mediation by CD4+ and CD8+ T cells and neutrophils expressing XCR1 receptor. AB - The C chemokine lymphotactin has been characterized as a T cell chemoattractant both in vitro and in vivo. To determine whether lymphotactin expression within tumors could influence tumor growth, we transfected an expression vector for lymphotactin into SP2/0 myeloma cells and tested their ability to form tumors in BALB/c and nude mice. Transfection did not alter cell growth in vitro. Whereas SP2/0 cells gave rise to a 100% tumor incidence, lymphotactin-expressing SP2/0 Lptn tumors invariably regressed in BALB/c mice and became infiltrated with CD4(+) and CD8(+) T cells and neutrophils. Regression of the SP2/0-Lptn tumors was associated with a type 1 cytokine response and dependent on both CD4(+) and CD8(+) T cells, but not NK cells. Both SP2/0 and SP2/0-Lptn tumors grew in nude mice, but growth of the latter tumors was retarded and associated with heavy neutrophil responses; this retardation of SP2/0-Lptn tumor growth was reversed by neutrophil depletion of the mice. Our data also indicate that mouse neutrophils express the lymphotactin receptor XCR1 and that lymphotactin specifically chemoattracts these cells in vitro. Thus, lymphotactin has natural adjuvant activities that may augment antitumor responses via effects on both T cells and neutrophils and thereby could be important in gene transfer immunotherapies for some cancers. PMID- 11418633 TI - Novel roles of CpG oligodeoxynucleotides as a leader for the sampling and presentation of CpG-tagged antigen by dendritic cells. AB - Oligodeoxynucleotides containing CpG motifs have been highlighted as potent Th1 activators. We previously reported that Ag and CpG, when conjugated together, synergistically promoted the Ag-specific Th1 development and inhibited the Th2 mediated airway eosinophilia. In this study, we examined the mechanisms underlying the synergism of the covalent conjugation. The CpG-OVA conjugate enhanced the Th1 activation and development. These characteristic features of the conjugate could not be ascribed to the polymerization of OVA, but mirrored the augmented binding of the CpG-tagged Ag to dendritic cells (DCs) in a CpG-guided manner, because phycobiliprotein, R-PE, conjugated to CpG stained a higher proportion of DCs with higher intensity than the mixture. R-PE fluorescence was emitted from cytoplasmic portions of the DCs, which simultaneously expressed costimulatory molecules and IL-12. The CpG-conjugated R-PE trafficking described above actually served as a potent Ag. These results indicate that CpG conjugated to Ag exhibit novel joint properties as promoters of Ag uptake and DC activators, thereby potentiating the ability of DCs to generate Th1 cells. The DNA-mediated promotion of Ag uptake would be advantageous for evoking host immune responses against invading microorganisms. PMID- 11418634 TI - Nitric oxide inhibits the transcription repressor Yin-Yang 1 binding activity at the silencer region of the Fas promoter: a pivotal role for nitric oxide in the up-regulation of Fas gene expression in human tumor cells. AB - NO has been increasingly implicated in control of the transcriptional machinery and serves as an intracellular second messenger to modify gene expression. We have demonstrated that NO up-regulated Fas receptor expression in ovarian carcinoma cell lines, albeit the mechanism involved is not known. Thus, we hypothesized that NO, directly or indirectly, may modify the transcriptional machinery that is responsible for the increased expression of the Fas gene. We examined the effect of NO on Fas gene expression using a Fas promoter-driven luciferase reporter system. Transient transfection of AD10 cells with pGL-3-FasP demonstrated that the IFN-gamma-dependent NO generation increases the trans activation of the Fas promoter, and this increase was blocked by the NOS inhibitor (N(G)-monomethyl-L-arginine), but could be restored by the addition of the NO donor S-nitroso-N-acetylpenicillamine. Systematic deletion of the Fas promoter revealed that the functional region responsible for the NO-mediated effect was located at the silencer region, suggesting that NO may be responsible for the disruption of a repressor mechanism. We demonstrate that NO up-regulates the expression of the Fas receptor on AD10 cells via the specific inactivation of the transcription repressor yin-yang 1 DNA binding activity to the silencer region of the Fas promoter. These findings reveal a new mechanism of NO-mediated gene regulation by interfering with a repressor transcription factor at the silencer region of the Fas promoter. PMID- 11418635 TI - Fas aggregation does not correlate with Fas-mediated apoptosis. AB - Cross-linking of cell surface Fas molecules by Fas ligand or by agonistic anti Fas Abs induces cell death by apoptosis. We found that a serine protease inhibitor, N-tosyl-L-lysine chloromethyl ketone (TLCK), dramatically enhances Fas mediated apoptosis in the human T cell line Jurkat and in various B cell lines resistant to Fas-mediated apoptosis. The enhancing effect of TLCK is specific to Fas-induced cell death, with no effect seen on TNF-alpha or TNF-related apoptosis inducing ligand-induced apoptosis. TLCK treatment had no effect on Fas expression levels on the cell surface, and neither promoted death-inducing signaling complex formation nor decreased expression levels of cellular inhibitors of apoptosis (FLICE inhibitory protein, X chromosome-linked inhibitor of apoptosis, and Bcl 2). Activation of the Fas-mediated apoptotic pathway by anti-Fas Ab is accompanied by aggregation of Fas molecules to form oligomers that are stable to boiling in SDS and beta-ME. Fas aggregation is often considered to be required for Fas-mediated apoptosis. However, sensitization of cells to Fas-mediated apoptosis by TLCK or other agents (cycloheximide, protein kinase C inhibitors) causes less Fas aggregation during the apoptotic process compared with that in nonsensitized cells. These results show that Fas aggregation and Fas-mediated apoptosis are not directly correlated and may even be inversely correlated. PMID- 11418636 TI - IL-1 enhances T cell-dependent antibody production through induction of CD40 ligand and OX40 on T cells. AB - IL-1 is a proinflammatory cytokine that plays pleiotropic roles in host defense mechanisms. We investigated the role of IL-1 in the humoral immune response using gene-targeted mice. Ab production against SRBC was significantly reduced in IL 1alpha/beta-deficient (IL-1(-/-)) mice and enhanced in IL-1R antagonist(-/-) mice. The intrinsic functions of T, B, and APCs were normal in IL-1(-/-) mice. However, we showed that IL-1(-/-) APCs did not fully activate DO11.10 T cells, while IL-1R antagonist (-/-) APCs enhanced the reaction, indicating that IL-1 promotes T cell priming through T-APC interaction. The function of IL-1 was CD28 CD80/CD86 independent. We found that CD40 ligand and OX40 expression on T cells was affected by the mutation, and the reduced Ag-specific B cell response in IL 1(-/-) mice was recovered by the treatment with agonistic anti-CD40 mAb both in vitro and in vivo. These observations indicate that IL-1 enhances T cell dependent Ab production by augmenting CD40 ligand and OX40 expression on T cells. PMID- 11418637 TI - Constitutive expression of MHC class II genes in melanoma cell lines results from the transcription of class II transactivator abnormally initiated from its B cell specific promoter. AB - In melanoma cell lines, two different patterns of MHC class II expression have been described, either an IFN gamma-inducible expression of HLA-DR and HLA-DP, with a faint or null expression of HLA-DQ, resembling that described for melanocytes, or a constitutive expression, i.e., IFN-gamma independent, of all three HLA-D isotypes. As this latter phenotype has been associated with a more rapid progression of melanoma tumors, we have analyzed in different melanoma cell lines the molecular mechanisms leading to this abnormal pattern of MHC class II expression. In agreement with the evidence of a coordinate transcription of the HLA-D genes in these cell lines, we have shown the constitutive expression of CIITA (class II transactivator) transcripts, CIITA being known as the master switch of MHC class II expression. Unexpectedly, these transcripts initiate from promoter III of the CIITA gene, a promoter that is mainly used constitutively in B lymphocytes. This expression was further shown to occur through factor(s) acting on the enhancer located upstream of CIITA promoter III, which was previously described in epithelioid cells as an IFN-gamma-response sequence. The hypothesis of a general abnormality of the IFN-gamma transduction pathway was dismissed. Constitutive transcription of CIITA from promoter III having been observed in unrelated melanoma cell lines, we propose the hypothesis that this phenomenon might not be a random event, but could be linked to the neoplasic state of the melanoma cells. PMID- 11418638 TI - Oral exposure to alloantigen generates intragraft CD8+ regulatory cells. AB - We have previously reported that oral administration of allogeneic rat spleen cells before kidney allotransplantation significantly prolongs graft survival. This prolongation was alloantigen specific and was associated with a decrease in graft-infiltrating cells (GIC) and an increase in transcription of IL-4 mRNA in the GIC. In this study increased splenic mixed lymphocyte responses from animals orally exposed to alloantigen before kidney transplantation suggested that the kidney allograft prolongation was not due to a masking of allorecognition, but to an immunomodulation of the immune response. We have assessed GIC T cell subsets on day 5 post-transplant and found decreased numbers of CD4(+) T cells in fed animals compared with controls, but there was no change in CD8(+) T cell numbers. The CD8(+) GIC from fed animals transcribed substantial levels of perforin, granzyme, and Fas ligand mRNA, indicating the presence of active CTL. Direct CTL assays showed that the GIC from fed recipients exhibited higher allo-CTL activity than GIC from control unfed recipients. In addition, the CD8(+) GIC exhibited high levels of IL-4 mRNA, suggesting Tc2-type regulatory cells. Prolonged graft survival in the face of active CTL and Tc2 cells suggests the presence of a CD8(+) regulatory cell population in the allograft. To confirm this, cell transfer experiments were performed. Prolongation of graft survival was transferred from rats orally exposed to alloantigen to naive animals by transfer of CD8(+) GIC. This is the first report that oral exposure to alloantigen prolongs kidney allograft survival by the generation of intragraft CD8(+) regulatory cells. PMID- 11418639 TI - Signal thresholds and modular synergy during expression of costimulatory molecules in B lymphocytes. AB - We analyzed intracellular pathways modulating surface densities of CD80 and CD86 in B cells activated through ligation of the Ag receptor, and the adhesion molecule CD54. Whereas B cell Ag receptor (BCR) cross-linking alone stimulated increased expression of CD86, up-regulation of CD80 required dual stimulation with anti-IgM and anti-CD54. The principal downstream component contributed by BCR signaling, toward both CD80 and CD86 induction, was the elevated concentration of free cytoplasmic Ca(2+), recruited by way of capacitative influx. This alone was sufficient to generate an increase in CD86 levels. However, CD80 enhancement required the concerted action of both intracellular Ca(2+) concentration and CD54-initiated pathways. The nexus between anti-IgM and anti-CD54 stimulation, in the context of CD80 regulation, was identified to involve a self-propagating process of sequential synergy. The first step involved amplified accumulation of intracellular cAMP, as a result of cross-talk between BCR-mobilized Ca(2+) and CD54-derived signals. This then facilitated a second synergistic interaction between Ca(2+) and cAMP, culminating in CD80 expression. Our findings of distinct signal transducer requirements, with the added consequences of cross-talk, offers an explanation for variable modulation of costimulatory molecule expression in response to diverse physiological stimuli. Importantly, these results also reveal how concentration threshold barriers for recruitment of individual second messengers can be overcome by constructive convergence of signaling modules. PMID- 11418640 TI - Antibody-induced shedding of CD44 from adherent cells is linked to the assembly of the cytoskeleton. AB - CD44 is a widely expressed integral membrane glycoprotein that serves as a specific adhesion receptor for the extracellular matrix glycosaminoglycan hyaluronan. CD44 participates in a variety of physiological and pathological processes through its role in cell adhesion. Under appropriate conditions, the ectodomain of CD44 is proteolytically removed from the cell surface. In this study we show that excessive CD44 shedding can be induced in mouse fibroblasts and monocytes upon exposure of these cells to a CD44-specific Ab immobilized on plastic, whereas treatment with phorbol ester induces significantly enhanced CD44 release from the monocytes only. CD44 shedding proceeds normally in fibroblasts and monocytes deficient in TNF-alpha converting enzyme (TACE), a sheddase involved in the processing of several substrates. Conversely, activation of the CD44 protease has no effect on the release of TNF-alpha from TACE-expressing cells, although the same metalloprotease inhibitor effectively blocks both TACE and the CD44 sheddase. Concomitant with anti-CD44 Ab- or phorbol ester-induced CD44 shedding, dramatic changes are observed in cell morphology and the structure of the actin cytoskeleton. Disruption of actin assembly with cytochalasin reduces CD44 shedding, but not the release of TNF-alpha. Moreover, pharmacological activation of Rho family GTPases Rac1 and Cdc42, which regulate actin filament assembly into distinct cytoskeletal structures, has a profound effect on CD44 release. We conclude that the CD44 sheddase and TACE are distinct enzymes, and that Ab- and phorbol ester-enhanced cleavage of CD44 is controlled in a cell type dependent fashion by Rho GTPases through the cytoskeleton. PMID- 11418641 TI - Enhancement of CD8+ T cell responses by ICOS/B7h costimulation. AB - Although the recently identified ICOS/B7h costimulatory counterreceptors are critical regulators of CD4(+) T cell responses, their ability to regulate CD8(+) responses is unclear. Here we report using a tumor-rejection model that ectopic B7h expression can costimulate rejection by CD8(+) T cells in the absence of CD4(+) T cells. Although responses of naive T cells were significantly augmented by priming with B7h, B7h was surprisingly effective in mobilizing recall responses of adoptively transferred T cells. To explore why secondary responses of CD8(+) T cells were particularly enhanced by B7h, kinetics of ICOS up regulation, proliferative responses, and cytokine production were compared from both naive and rechallenged 2C-transgenic T cells costimulated in vitro. Although B7h costimulated proliferative responses from both CD8(+) populations, rechallenged cells were preferentially costimulated for IL-2 and IFN-gamma production. These results indicate that ICOS/B7h counterreceptors likely function in vivo to enhance secondary responses by CD8(+) T cells. PMID- 11418642 TI - CD28-independent costimulation of T cells in alloimmune responses. AB - T cell costimulation by B7 molecules plays an important role in the regulation of alloimmune responses. Although both B7-1 and B7-2 bind CD28 and CTLA-4 on T cells, the role of B7-1 and B7-2 signaling through CTLA-4 in regulating alloimmune responses is incompletely understood. To address this question, we transplanted CD28-deficient mice with fully allogeneic vascularized cardiac allografts and studied the effect of selective blockade of B7-1 or B7-2. These mice reject their grafts by a mechanism that involves both CD4(+) and CD8(+) T cells. Blockade of CTLA-4 or B7-1 significantly accelerated graft rejection. In contrast, B7-2 blockade significantly prolonged allograft survival and, unexpectedly, reversed the acceleration of graft rejection caused by CTLA-4 blockade. Furthermore, B7-2 blockade prolonged graft survival in recipients that were both CD28 and CTLA-4 deficient. Our data indicate that B7-1 is the dominant ligand for CTLA-4-mediated down-regulation of alloimmune responses in vivo and suggest that B7-2 has an additional receptor other than CD28 and CTLA-4 to provide a positive costimulatory signal for T cells. PMID- 11418643 TI - An immunodominant MHC class II-restricted tumor antigen is conformation dependent and binds to the endoplasmic reticulum chaperone, calreticulin. AB - There is accumulating evidence that CD4(+) T cell responses are important in antitumor immunity. Accordingly, we generated CD4(+) T cells against the murine CT26 colon cancer. Three of three independent CT26-specific CD4(+) hybridomas were found to recognize the high m.w. precursor of the env gene product gp90. The CD4(+) response was completely tumor specific in that the same glycoprotein expressed by other tumors was not recognized by the CT26-specific hybridomas. The recognition of gp90 by the hybridomas was strictly dependent on the conformation of gp90. Different procedures that disrupted the conformation of the glycoprotein, such as disulfide bond reduction and thermal denaturation, completely abrogated recognition of gp90 by all three hybridomas. In CT26 cells, but not in other tumor cells tested, a large proportion of gp90 was retained in the endoplasmic reticulum, mostly bound to the endoplasmic reticulum chaperone, calreticulin. Although calreticulin was not essential for the stimulation of the gp90-specific hybridomas, most of the antigenic form of gp90 was bound to it. The antigenicity of gp90 correlated well with calreticulin binding, reflecting the fact that specificity of binding of calreticulin to its substrate required posttranslational modifications that were also necessary for the generation of this tumor-specific CD4(+) epitope. PMID- 11418644 TI - IL-12 augments CD8+ T cell development for contact hypersensitivity responses and circumvents anti-CD154 antibody-mediated inhibition. AB - During sensitization with dinitrofluorobenzene for contact hypersensitivity (CHS) responses, hapten-specific CD8(+) T cells develop into IFN-gamma-producing cells, and CD4(+) T cells develop into IL-4/IL-5-producing cells. Administration of IL 12 during sensitization skews CD4(+) T cell development to IFN-gamma-producing cells, resulting in exaggerated CHS responses. In the current report we tested the role of IL-12 on CD8(+) T cell development during sensitization and elicitation of CHS to dinitrofluorobenzene. Administration of IL-12 during hapten sensitization induced the expression of IL-12Rbeta2 on both CD4(+) and CD8(+) T cells, augmented IFN-gamma production by these T cell populations, and increased the magnitude and duration of the CHS response to hapten challenge. CHS responses were virtually identical in wild-type and IL-12 p40(-/-) mice. Since engagement of CD40 on APC may stimulate IL-12 production, we also tested the role of CD40 CD154 interactions on the development of IFN-gamma-producing CD4(+) and CD8(+) T cells following hapten sensitization. Development of IFN-gamma-producing CD4(+) T cells during hapten sensitization was absent in wild-type mice treated with anti CD154 mAb or in CD154(-/-) mice. In contrast, the absence of CD40-CD154 signaling had little or no impact on the development of IFN-gamma-producing CD8(+) T cells. These results demonstrate that the development of hapten-specific Th1 effector CD4(+) T cells in CHS requires both CD40-CD154 interactions and IL-12, whereas the development of IFN-gamma-producing effector CD8(+) T cells can occur independently of these pathways. PMID- 11418645 TI - The role of the CD19/CD21 complex in B cell processing and presentation of complement-tagged antigens. AB - The CD19/CD21 complex is an essential B cell coreceptor that functions synergistically to enhance signaling through the B cell Ag receptor in response to T cell-dependent, complement-tagged Ags. In this study, we use a recombinant protein containing three tandemly arranged copies of C3d and the Ag hen egg lysozyme, shown to be a highly effective immunogen in vivo, to evaluate the role of the CD19/CD21 complex in Ag processing in B cells. Evidence is provided that coengagement of the CD19/CD21 complex results in more rapid and efficient production of antigenic peptide/class II complexes as compared with B cell Ag receptor-mediated processing alone. The CD19/CD21 complex does not itself target complement-tagged Ags for processing, but rather appears to influence B cell Ag processing through its signaling function. The ability of the CD19/CD21 complex to augment processing may be an important element of the mechanism by which the CD19/CD21 complex functions to promote B cell responses to T cell-dependent complement-tagged Ags in vivo. PMID- 11418646 TI - TNF-alpha-induced sphingosine 1-phosphate inhibits apoptosis through a phosphatidylinositol 3-kinase/Akt pathway in human hepatocytes. AB - Human hepatocytes usually are resistant to TNF-alpha cytotoxicity. In mouse or rat hepatocytes, repression of NF-kappaB activation is sufficient to induce TNF alpha-mediated apoptosis. However, in both Huh-7 human hepatoma cells and Hc human normal hepatocytes, when infected with an adenovirus expressing a mutated form of IkappaBalpha (Ad5IkappaB), which almost completely blocks NF-kappaB activation, >80% of the cells survived 24 h after TNF-alpha stimulation. Here, we report that TNF-alpha activates other antiapoptotic factors, such as sphingosine kinase (SphK), phosphatidylinositol 3-kinase (PI3K), and Akt kinase. Pretreatment of cells with N,N-dimethylsphingosine (DMS), an inhibitor of SphK, or LY 294002, an inhibitor of PI3K that acts upstream of Akt, increased the number of apoptotic cells induced by TNF-alpha in Ad5IkappaB-infected Huh-7 and Hc cells. TNF-alpha induced activations of PI3K and Akt were inhibited by DMS. In contrast, exogenous sphingosine 1-phosphate, a product of SphK, was found to activate Akt and partially rescued the cells from TNF-alpha-induced apoptosis. Although Akt has been reported to activate NF-kappaB, DMS and LY 294002 failed to prevent TNF alpha-induced NF-kappaB activation, suggesting that the antiapoptotic effects of SphK and Akt are independent of NF-kappaB. Furthermore, apoptosis mediated by Fas ligand (FasL) involving Akt activation also was potentiated by DMS pretreatment in Hc cells. Sphingosine 1-phosphate administration partially protected cells from FasL-mediated apoptosis. These results indicate that not only NF-kappaB but also SphK and PI3K/Akt are involved in the signaling pathway(s) for protection of human hepatocytes from the apoptotic action of TNF-alpha and probably FasL. PMID- 11418647 TI - Functional heterogeneity of cytokines and cytolytic effector molecules in human CD8+ T lymphocytes. AB - CD8(+) T cells use a number of effector mechanisms to protect the host against infection. We have studied human CD8(+) T cells specific for CMV pp65(495-503) epitope, or for staphylococcal enterotoxin B, for the expression patterns of five cytokines and cytolytic effector molecules before and after antigenic stimulation. Ex vivo, the cytolytic molecule granzyme B was detected in a majority of circulating CMV-specific CD8(+) T cells, whereas perforin was rarely expressed. Both were highly expressed after Ag-specific activation accompanied by CD45RO up-regulation. TNF-alpha, IFN gamma, and IL-2 were sequentially acquired on recognition of Ag, but surprisingly, only around half of the CMV-specific CD8(+) T cells responded to antigenic stimuli with production of any cytokine measured. A dominant population coexpressed TNF-alpha and IFN-gamma, and cells expressing TNF-alpha only, IFN-gamma only, or all three cytokines together also occurred at lower but clearly detectable frequencies. Interestingly, perforin expression and production of IFN-gamma and TNF-alpha in CD8(+) T cells responding to staphylococcal enterotoxin B appeared to be largely segregated, and no IL-2 was detected in perforin-positive cells. Together, these data indicate that human CD8(+) T cells can be functionally segregated in vivo and have implications for the understanding of human CD8(+) T cell differentiation and specialization and regulation of effector mechanisms. PMID- 11418648 TI - Generation of anergic and potentially immunoregulatory CD25+CD4 T cells in vivo after induction of peripheral tolerance with intravenous or oral antigen. AB - Immunoregulatory CD25(+)CD4 T cells are thought to arise from the thymus as a distinct lineage of CD4 T cells specific for self Ags. We used the DO11.10 TCR transgenic adoptive transfer system to show that cells of similar phenotype may also arise in the course of peripheral tolerance induction. Such cells emerged within 1 wk following Ag exposure and correlated negatively with the number of initial cell divisions. Limiting i.v. Ag dose or using an oral tolerance protocol yielded the greatest numbers of Ag-specific CD25(+)CD4 T cells. In contrast, immunogenic Ag exposure in the presence of an adjuvant did not lead to emergence of CD25(+)CD4 T cells. The profound anergic phenotype of these cells and their potential immunoregulatory properties make them an especially desirable population to induce in the course of immunotherapy in numerous clinical settings. This experimental system may be useful in future studies designed to optimize immunologic tolerance induction. PMID- 11418649 TI - Rapid induction of naive T cell apoptosis by ecto-nicotinamide adenine dinucleotide: requirement for mono(ADP-ribosyl)transferase 2 and a downstream effector. AB - Lymphocytes express a number of NAD-metabolizing ectoenzymes, including mono(ADP ribosyl)transferases (ART) and ADP ribosylcyclases. These enzymes may regulate lymphocyte functions following the release of NAD in injured or inflammatory tissues We report here that extracellular NAD induces apoptosis in BALB/c splenic T cells with an IC(50) of 3-5 microM. Annexin V staining of cells was observed already 10 min after treatment with NAD in the absence of any additional signal. Removal of GPI-anchored cell surface proteins by phosphatidylinositol-specific phospholipase C treatment rendered cells resistant to NAD-mediated apoptosis. RT PCR analyses revealed that resting BALB/c T cells expressed the genes for GPI anchored ART2.1 and ART2.2 but not ART1. ART2-specific antisera blocked radiolabeling of cell surface proteins with both [(32)P]NAD and NAD-mediated apoptosis. Further analyses revealed that natural knockout mice for Art2.a (C57BL/6) or Art2.b (NZW) were resistant to NAD-mediated apoptosis. Labeling with [(32)P]NAD revealed strong cell surface ART activity on T cells of C57BL/6 and little if any activity on cells of NZW mice. T cells of (C57BL/6 x NZW)F(1) animals showed strong cell surface ART activity and were very sensitive to NAD induced apoptosis. As in BALB/c T cells, ART2-specific antisera blocked cell surface ART activity and apoptosis in (C57BL/6 x NZW)F(1) T cells. The fact that T cells of F(1) animals are sensitive to rapid NAD-induced apoptosis suggests that this effect requires the complementation of (at least) two genetic components. We propose that one of these is cell surface ART2.2 activity (defective in the NZW parent), the other a downstream effector of ADP ribosylation (defective in the C57BL/6 parent). PMID- 11418650 TI - Partially distinct molecular mechanisms mediate inhibitory FcgammaRIIB signaling in resting and activated B cells. AB - FcgammaRIIB functions as an inhibitory receptor to dampen B cell Ag receptor signals and immune responses. Accumulating evidence indicates that ex vivo B cells require the inositol 5-phosphatase, Src homology domain 2-containing inositol 5-phosphatase (SHIP), for FcgammaRIIB-mediated inhibitory signaling. However, we report here that LPS-activated primary B cells do not require SHIP and thus differ from resting B cells. SHIP-deficient B cell blasts display efficient FcgammaRIIB-dependent inhibition of calcium mobilization as well as Akt and extracellular signal-related protein kinase phosphorylation. Surprisingly, FcgammaRIIB-dependent degradation of phosphatidylinositol 3,4,5-trisphosphate and conversion into phosphatidylinositol 3,4-bisphosphate occur in SHIP-deficient B cell blasts, demonstrating the function of an additional inositol 5-phosphatase. Further analysis reveals that while resting cells express only SHIP, B cell blasts also express the recently described inositol 5-phosphatase, SHIP-2. Finally, data suggest that both SHIP-2 and SHIP can mediate downstream biologic consequences of FcgammaRIIB signaling, including inhibition of the proliferative response. PMID- 11418651 TI - Memory T cells constitute a subset of the human CD8+CD45RA+ pool with distinct phenotypic and migratory characteristics. AB - Using HLA class I-viral epitope tetramers to monitor herpes virus-specific CD8(+) T cell responses in humans, we have shown that a significant fraction of responding cells revert from a CD45RO(+) to a CD45RA(+) state after priming. All tetramer-binding CD45RA(+) cells, regardless of epitope specificity, expressed a phenotype LFA-1(high)CCR7(low) that was stable for at least 10 years in infectious mononucleosis patients and indefinitely in asymptomatic carriers. CD8(+)CD45RA(+)LFA-1(high) cells were not present in cord blood but in adults account for up to 50% of CD8(+)CD45RA(+) cells. These CD45RA(+)LFA-1(high) cells have significantly shorter telomeres than CD45RA(+)LFA-1(low) cells, suggesting that the latter represent a naive population, while the former are memory cells. CD45RA(+) memory cells are a stable population of noncycling cells, but on stimulation they are potent producers of IFN-gamma, while naive CD8(+) cells produce only IL-2. The chemokine receptor profile and migratory potential of CD45RA(+) memory cells is very similar to CD45RO(+) cells but different to naive CD8 cells. In accord with this, CD45RA(+) memory cells were significantly underrepresented in lymph nodes, but account for virtually all CD8(+)CD45RA(+) T cells in peripheral tissues of the same individuals. PMID- 11418652 TI - Positive regulatory role of IL-12 in macrophages and modulation by IFN-gamma. AB - Similar to myeloid dendritic cells, murine macrophages and macrophage cell lines were found to express a surface receptor for IL-12. As a result, peritoneal macrophages could be primed by IL-12 to present an otherwise poorly immunogenic tumor peptide in vivo. Using binding analysis and RNase protection assay, we detected a single class of high affinity IL-12 binding sites (K(d) of approximately 35 pM) whose number per cell was increased by IFN-gamma via up regulation of receptor subunit expression. Autocrine production of IL-12 was suggested to be a major effect of IL-12 on macrophages when the cytokine was tested alone or after priming with IFN-gamma in vitro. In vivo, combined treatment of macrophages with IFN-gamma and IL-12 resulted in synergistic effects on tumor peptide presentation. Therefore, our findings suggest a general and critical role of IL-12 in potentiating the accessory function of myeloid APC. PMID- 11418653 TI - Regulation of dendritic cell recruitment into resting and inflamed airway epithelium: use of alternative chemokine receptors as a function of inducing stimulus. AB - Dendritic cells (DC) were purified by flow cytometry from rat tracheal mucosa; they exhibited the phenotypic characteristics of immature DC including high endocytic activity, low CD80/86 expression, and in vitro responsiveness to a broad range of CC chemokines. Daily treatment of adult rats with the selective CCR1 and CCR5 antagonist Met-RANTES reduced baseline numbers of tracheal intraepithelial DC by 50-60%, and pretreatment of animals with Met-RANTES before inhalation of aerosol containing heat-killed bacteria abolished the rapid DC influx into the epithelium that occurred in untreated controls, implicating CCR1 and CCR5 and their ligands in recruitment of immature DC precursors into resting airway tissues and during acute bacterial-induced inflammation. Comparable levels of DC recruitment were observed during airway mucosal Sendai virus infection and after aerosol challenge of sensitized animals with the soluble recall Ag OVA. However, Met-RANTES did not affect these latter responses, indicating the use of alternative chemokine receptors/ligands for DC recruitment, or possibly attraction of different DC subsets, depending on the nature of the eliciting stimulus. PMID- 11418654 TI - Down-regulation of IL-12 p40 gene in Plasmodium berghei-infected mice. AB - We analyzed the mechanism that causes suppression of IL-12 p40 gene induction during Plasmodium berghei infection. Although IL-12 together with IFN-gamma plays an important role in protection against pathogenic infection, the IL-12 p70 protein production of infected macrophages is lower than that by the uninfected macrophages. We showed in the present study that the induction of IL-12 p40 gene but not IL-12 p35 gene in macrophages of P. berghei-infected mice was profoundly inhibited. The inhibition was induced by interaction with macrophages that had contacted with P. berghei-infected erythrocytes and was mediated by a soluble factor, IL-10. There was comparable activation of NF-kappaB in uninfected and infected cells. The induction of IFN-regulatory factor-1 gene was comparable in transcription level in uninfected and infected cells, while the unidentified complex formation of IFN-regulatory factor-1 was observed in infected cells. Therefore, the inhibition of the IL-12 p40 gene induction appeared to be regulated at transcriptional regulation level of the gene. PMID- 11418655 TI - B-Myb overexpression results in activation and increased Fas/Fas ligand-mediated cytotoxicity of T and NK cells. AB - The human B-myb gene encodes a transcriptional regulator that plays an important role in cell cycle progression, differentiation, and survival. To assess the in vivo role of B-myb, we investigated the phenotype of mouse transgenic lines in which B-Myb expression in lymphoid tissues was driven by the LCK proximal promoter. Overexpression of B-Myb had no measurable effect on the subsets of splenic and thymic lymphocytes, but was associated with increased expression of Fas ligand in NK and T cells. B-Myb-overexpressing splenocytes expressed higher IFN-gamma levels and contained higher percentages of cytokine-producing cells than wild-type (wt) splenocytes, as detected by Western blot analysis and ELISPOT assays, respectively. Ex vivo-cultured transgenic thymocytes and splenocytes had decreased survival compared with the corresponding cells from wt mice, possibly dependent on increased expression of Fas ligand. In addition, Fas ligand dependent cytotoxicity of transgenic T and NK cells was significantly higher than that mediated by their wt counterparts. Together, these results indicate that B Myb overexpression results in T and NK cell activation and increased cytotoxicity. Therefore, in addition to its well-established role in proliferation and differentiation, B-myb also appears to be involved in activation of NK and T cells and in their regulation of Fas/Fas ligand-mediated cytotoxicity PMID- 11418656 TI - Pertussis toxin inhibits induction of tissue-specific autoimmune disease by disrupting G protein-coupled signals. AB - Pertussis toxin (PTX) has been used for many years as an adjuvant that promotes development of tissue-specific experimental autoimmune diseases such as experimental autoimmune encephalomyelitis, experimental autoimmune uveitis (EAU), and others. Enhancement of vascular permeability and of Th1 responses have been implicated in this effect. Here we report a surprising observation that, in a primed system, PTX can completely block the development of EAU. Disease was induced in B10.RIII mice by adoptive transfer of uveitogenic T cells, or by immunization with a uveitogenic peptide. A single injection of PTX concurrently with infusion of the uveitogenic T cells, or two injections 7 and 10 days after active immunization, completely blocked development of EAU. EAU also was prevented by a 1-h incubation in vitro of the uveitogenic T cells with PTX before infusing them into recipients. Uveitogenic T cells treated with PTX in vitro and lymphoid cells from mice treated with PTX in vivo failed to migrate to chemokines in a standard chemotaxis assay. Neither the isolated B-oligomer subunit of PTX that lacks ADP ribosyltransferase activity nor the related cholera toxin that ADP ribosylates G(s) (but not G(i)) proteins blocked EAU induction or migration to chemokines. We conclude that PTX present at the time of cell migration to the target organ prevents EAU, and propose that it does so at least in part by disrupting signaling through G(i) protein-coupled receptors. Thus, the net effect of PTX on autoimmune disease would represent an integration of enhancing and inhibitory effects. PMID- 11418657 TI - Unequal VH gene rearrangement frequency within the large VH7183 gene family is not due to recombination signal sequence variation, and mapping of the genes shows a bias of rearrangement based on chromosomal location. AB - Much of the nonrandom usage of V, D, and J genes in the Ab repertoire is due to different frequencies with which gene segments undergo V(D)J rearrangement. The recombination signal sequences flanking each segment are seldom identical with consensus sequences, and this natural variation in recombination signal sequence (RSS) accounts for some differences in rearrangement frequencies in vivo. Here, we have sequenced the RSS of 19 individual V(H)7183 genes, revealing that the majority have one of two closely related RSS. One group has a consensus heptamer, and the other has a nonconsensus heptamer. In vitro recombination substrate studies show that the RSS with the nonconsensus heptamer, which include the frequently rearranging 81X, rearrange less well than the RSS with the consensus heptamer. Although 81X differs from the other 7183-I genes at three positions in the spacer, this does not significantly increase its recombination potency in vitro. The rearrangement frequency of all members of the family was determined in microMT mice, and there was no correlation between the in vitro recombination potential and V(H) gene rearrangement frequency in vivo. Furthermore, genes with identical RSS rearrange at different frequencies in vivo. This demonstrates that other factors can override differences in RSS potency in vivo. We have also determined the gene order of all V(H)7183 genes in a bacterial artificial chromosome contig and show that most of the frequently rearranging genes are in the 3' half of the region. This suggests that chromosomal location plays an important role in nonrandom rearrangement of the V(H)7183 genes. PMID- 11418658 TI - Transcription factor NF-kappa B regulates Ig lambda light chain gene rearrangement. AB - The tissue- and stage-specific assembly of Ig and TCR genes is mediated by a common V(D)J recombinase complex in precursor lymphocytes. Directed alterations in the accessibility of V, D, and J gene segments target the recombinase to specific Ag receptor loci. Accessibility within a given locus is regulated by the functional interaction of transcription factors with cognate enhancer elements and correlates with the transcriptional activity of unrearranged gene segments. As demonstrated in our prior studies, rearrangement of the Igkappa locus is regulated by the inducible transcription factor NF-kappaB. In contrast to the Igkappa locus, known transcriptional control elements in the Iglambda locus lack functional NF-kappaB binding sites. Consistent with this observation, the expression of assembled Iglambda genes in mature B cells has been shown to be NF kappaB independent. Nonetheless, we now show that specific repression of NF kappaB inhibits germline transcription and recombination of Iglambda gene segments in precursor B cells. Molecular analyses indicate that the block in NF kappaB impairs Iglambda rearrangement at the level of recombinase accessibility. In contrast, the activities of known Iglambda promoter and enhancer elements are unaffected in the same cellular background. These findings expand the range of NF kappaB action in precursor B cells beyond Igkappa to include the control of recombinational accessibility at both L chain loci. Moreover, our results strongly suggest the existence of a novel Iglambda regulatory element that is either directly or indirectly activated by NF-kappaB during the early stages of B cell development. PMID- 11418659 TI - Critical role for CD8 in binding of MHC tetramers to TCR: CD8 antibodies block specific binding of human tumor-specific MHC-peptide tetramers to TCR. AB - There are conflicting opinions about the role that the T cell coreceptors CD4 and CD8 play in TCR binding and activation. Recent evidence from transgenic mouse models suggests that CD8 plays a critical role in TCR binding and activation by peptide-MHC complex multimers (tetramers). Here we show with a human CTL clone specific for a tumor-associated MHC-peptide complex that the binding of tetramers to the TCR on these cells is completely blocked by anti-human CD8 Abs. Moreover, the staining of CTLs with specific MHC-peptide tetramers simultaneously with anti CD8 Abs was completely blocked with three different anti-CD8 Abs. This blockage was mediated by anti-CD8 Abs but not anti-CD3 Abs and was dose dependent. The blocking effect of the anti-CD8 Abs was attributable to directly inhibiting tetramer binding and was not attributable to Ab-mediated TCR-CD8 internalization and down-regulation. Our results have important implications in TCR binding to MHC-peptide tetramers. MHC-peptide tetramers are widely used today in combination with anti-CD8 Abs for the phenotypic analysis of T cell populations and in the study of T cell responses under various pathological conditions such as infectious diseases and cancer. Our results indicate that also in the human system CD8 plays a critical role in the interaction of MHC-peptide multimers with TCR. PMID- 11418660 TI - The Sushi domain of soluble IL-15 receptor alpha is essential for binding IL-15 and inhibiting inflammatory and allogenic responses in vitro and in vivo. AB - IL-15 is a pleiotropic cytokine that plays important roles in both innate and adaptive immunity. It is associated with a range of immunopathology, including rheumatoid arthritis and allograft rejection. IL-15 functions through the trimeric IL-15R complex, which consists of a high affinity binding alpha-chain and the common IL-2R beta- and gamma-chains. Characterization of IL-15/IL-15R interactions may facilitate the development of improved IL-15 antagonists for therapeutic interventions. We previously constructed soluble murine IL-15Ralpha (sIL-15Ralpha) by deleting the cytoplasmic and transmembrane domains. To localize the functional domain of IL-15Ralpha, we have now constructed various truncated versions of sIL-15Ralpha. The shortest region retaining IL-15 binding activity is a 65-aa sequence spanning the Sushi domain of IL-15Ralpha. Sushi domains, common motifs in protein-protein interactions, contain four cysteines forming two disulfide bonds in a 1-3 and 2-4 pattern. Amino acid substitution of the first or fourth cysteine in sIL-15Ralpha completely abolished its IL-15 binding activity. This also abrogated the ability of sIL-15Ralpha to neutralize IL-15-induced proinflammatory cytokine production and anti-apoptotic response in vitro. Furthermore, the mutant sIL-15Ralpha lost its ability to inhibit carrageenan induced local inflammation and allogenic cell-induced T cell proliferation and cytokine production in vivo. Thus, the Sushi domain is critical for the functional activity of sIL-15Ralpha. PMID- 11418661 TI - IL-1 induced chemokine production through the association of Syk with TNF receptor-associated factor-6 in nasal fibroblast lines. AB - The fibroblasts stimulated by cytokines released the chemokine and recruited the infiltrating cells, including eosinophils, that play a key role in the pathogenesis of airway disease. We established the human fibroblast lines showing high Syk expression and the lines showing low Syk expression from pieces of nasal polyp. IL-1 induces the interaction of TNFR-associated factor (TRAF) 6 with IL-1R associated kinase, which is rapidly recruited to the IL-1R after IL-1 induction, whereas TRAF2 participates in TNF-alpha-signaling. In the present study, we found that Syk played a different role in IL-1- and TNF-alpha-induced chemokine production through a signaling complex involving Syk and TRAF6. Overexpression of wild-type Syk by gene transfer enhanced RANTES production from nasal fibroblasts stimulated with IL-1. The decrease of Syk expression by the administration of Syk antisense inhibited RANTES production in response to IL-1. However, the change of Syk expression did not affect RANTES production by TNF-alpha stimulation. We concluded that Syk is required for the IL-1-induced chemokine production through the association with TRAF-6 in fibroblasts of nasal polyps. PMID- 11418662 TI - Human Notch-1 inhibits NF-kappa B activity in the nucleus through a direct interaction involving a novel domain. AB - Notch participates in diverse cell fate decisions throughout embryonic development and postnatal life. Members of the NF-kappaB/Rel family of transcription factors are involved in the regulation of a variety of genes important for immune function. The biological activity of the NF-kappaB transcription factors is controlled by IkappaB proteins. Our previous work demonstrated that an intracellular, constitutively active form of human Notch 1/translocation-associated Notch homologue-1 (Notch(IC)) functions as an IkappaB molecule with specificity for the NF-kappaB p50 subunit and physically interacts with NF-kappaB in T cells. In the current study, we investigated the roles of different domains of Notch(IC) in the regulation of NF-kappaB-directed gene expression and NF-kappaB DNA binding activity. We found that Notch(IC) localizes to the nucleus and that a region in the N-terminal portion of Notch(IC), not the six ankyrin repeats, is responsible for the inhibitory effects of Notch on NF kappaB-directed gene expression and NF-kappaB DNA binding activity. The N terminal portion of Notch(IC) inhibited p50 DNA binding and interacted specifically with p50 subunit, not p65 of NF-kappaB. The interaction between Notch and NF-kappaB indicates that in addition to its role in the development of the immune system, Notch-1 may also have critical functions in the immune response, inflammation, viral infection, and apoptosis through control of NF kappaB-mediated gene expression. PMID- 11418663 TI - Structural effects of framework mutations on a humanized anti-lysozyme antibody. AB - A humanized version of the mouse anti-lysozyme Ab D1.3 was previously constructed as an Fv fragment and its structure was crystallographically determined in the free form and in complex with lysozyme. Here we report five new crystal structures of single-amino acid substitution mutants of the humanized Fv fragment, four of which were determined as Fv-lysozyme complexes. The crystals were isomorphous with the parent forms, and were refined to free R values of 28 31% at resolutions of 2.7-2.9 A. Residue 27 in other Abs has been implicated in stabilizing the conformation of the first complementarity-determining region (CDR) of the H chain, residues 31-35. We find that a Phe-to-Ser mutation at 27 alters the conformation of immediately adjacent residues, but this change is only weakly transmitted to Ag binding residues in the nearby CDR. Residue 71 of the H chain has been proposed to control the relative disposition of H chain CDRs 1 and 2, based on the bulk of its side chain. However, in structures we determined with Val, Ala, or Arg substituted in place of Lys at position 71, no significant change in the conformation of CDRs 1 and 2 was observed. PMID- 11418664 TI - A NF-kappa B/Sp1 region is essential for chromatin remodeling and correct transcription of a human granulocyte-macrophage colony-stimulating factor transgene. AB - The GM-CSF gene is expressed following activation of T cells. The proximal promoter and an upstream enhancer have previously been characterized using transfection and reporter assays in T cell lines in culture. A 10.5-kb transgene containing the entire human GM-CSF gene has also been shown to display inducible, position-independent, copy number-dependent transcription in mouse splenocytes. To determine the role of individual promoter elements in transgene function, mutations were introduced into the proximal promoter and activity assessed following the generation of transgenic mice. Of four mutations introduced into the transgene promoter, only one, in an NF-kappaB/Sp1 region, led to decreased induction of the transgene in splenocytes or bone marrow-derived macrophages. This mutation also affected the activity of reporter gene constructs stably transfected into T cell lines in culture, but not when transiently transfected into the same cell lines. The mutation alters the NF-kappaB family members that bind to the NF-kappaB site as well as reducing the binding of Sp1 to an adjacent element. A DNase I hypersensitive site that is normally generated at the promoter following T cell activation on the wild-type transgene does not appear in the mutant transgene. These results suggest that the NF-kappaB/Sp1 region plays a critical role in chromatin remodeling and transcription on the GM-CSF promoter in primary T cells. PMID- 11418665 TI - The Smad3 protein is involved in TGF-beta inhibition of class II transactivator and class II MHC expression. AB - TGF-beta is a immunoregulatory cytokine that inhibits class II MHC expression in a variety of cell types. Previous studies have shown that the class II MHC transactivator (CIITA), a master regulator that controls class II MHC expression, is targeted by TGF-beta for repression of IFN-gamma-induced class II MHC expression in astrocytes. The mechanism(s) underlying the TGF-beta inhibitory effect is not understood. In this study, we demonstrate that TGF-beta inhibition of CIITA expression occurs at the transcriptional level, and that both constitutive and IFN-gamma-induced human CIITA type IV promoter activity is inhibited by TGF-beta. TGF-beta does not affect the signaling events that mediate IFN-gamma activation of CIITA expression; i.e, TGF-beta does not inhibit IFN gamma-induced STAT-1alpha phosphorylation and/or DNA binding ability, nor is IFN gamma induction of IFN regulatory factor affected. The inhibitory effect of TGF beta on the type IV CIITA promoter is mediated through a promoter region within 80 bp from the transcription start site. Elimination of TGF-beta inhibition of class II MHC and CIITA expression in Smad3-deficient astrocytes, as well as restoration of the inhibitory effect by overexpression of the Smad3 protein, demonstrates that Smad3 is essential in mediating TGF-beta inhibition of CIITA and class II MHC expression. PMID- 11418666 TI - Induction of germline transcription in the human TCR gamma locus by STAT5. AB - TCR and Ig genes are assembled by V(D)J recombination during lymphocyte development. The enhancer and the germline promoter control the accessibility of each locus for the common recombinase activity. In the mouse TCRgamma locus, STAT5 proteins activated by the IL-7R interact with consensus motifs in 5' regions of Jgamma segments and induce germline transcription. To evaluate the role of STAT5 in controlling the accessibility of the TCRgamma locus, we characterized the germline transcription of human TCRgamma genes and compared it with mouse. We first demonstrated that Jgamma-Cgamma germline transcripts are induced in a cytokine-dependent human erythroleukemia cell line. STAT consensus motifs are present in 5' regions of Jgamma1.1 and Jgamma2.1 gene segments, and activated STAT5 binds to these motifs. By using a reporter assay, we showed that the Jgamma1.1 germline promoter is transactivated by STAT5 and that mutations in any of the two STAT motifs abrogate this activity. Thus, this study demonstrates that STAT5 induces germline transcription in the TCRgamma locus of both mouse and human and suggests the possibility that this mechanism may play an essential role in controlling the TCRgamma locus accessibility. In addition, STAT motifs are conserved among 5' Jgamma germline promoters, 3' enhancers, and a locus control region-like element, HsA, in both mouse and human TCRgamma loci, indicating the possibility that IL-7R/STAT5 signaling probably controls the locus-wide accessibility through these elements. PMID- 11418667 TI - Decreased frequency of somatic hypermutation and impaired affinity maturation but intact germinal center formation in mice expressing antisense RNA to DNA polymerase zeta. AB - To examine a role of DNA polymerase zeta in somatic hypermutation, we generated transgenic mice that express antisense RNA to a portion of mouse REV3, the gene encoding this polymerase. These mice express high levels of antisense RNA, significantly reducing the levels of endogenous mouse REV3 transcript. Following immunization to a hapten-protein complex, transgenic mice mounted vigorous Ab responses, accomplished the switch to IgG, and formed numerous germinal centers. However, in most transgenic animals, the generation of high affinity Abs was delayed. In addition, accumulation of somatic mutations in the V(H) genes of memory B cells from transgenic mice was decreased, particularly among those that generate amino acid replacements that enhance affinity of the B cell receptor to the hapten. These data implicate DNA polymerase zeta, a nonreplicative polymerase, in the process of affinity maturation, possibly through a role in somatic hypermutation, clonal selection, or both. PMID- 11418668 TI - Human thymic stromal lymphopoietin preferentially stimulates myeloid cells. AB - The sequence of a novel hemopoietic cytokine was discovered in a computational screen of genomic databases, and its homology to mouse thymic stromal lymphopoietin (TSLP) suggests that it is the human orthologue. Human TSLP is proposed to signal through a heterodimeric receptor complex that consists of a new member of the hemopoietin family termed human TSLP receptor and the IL-7R alpha-chain. Cells transfected with both receptor subunits proliferated in response to purified, recombinant human TSLP, with induced phosphorylation of Stat3 and Stat5. Human TSLPR and IL-7Ralpha are principally coexpressed on monocytes and dendritic cell populations and to a much lesser extent on various lymphoid cells. In accord, we find that human TSLP functions mainly on myeloid cells; it induces the release of T cell-attracting chemokines from monocytes and, in particular, enhances the maturation of CD11c(+) dendritic cells, as evidenced by the strong induction of the costimulatory molecules CD40 and CD80 and the enhanced capacity to elicit proliferation of naive T cells. PMID- 11418669 TI - Unstimulated human CD4 lymphocytes express a cytoplasmic immature form of the common cytokine receptor gamma-chain. AB - As a component of various cytokine receptors, common cytokine receptor gamma chain (gamma(c)) is essential in the development of the immune system and plays an important role in different stages of inflammatory and immune responses. Here we establish that resting CD4 T cells and the Jurkat CD4 T cell line do not express the mature form of gamma(c) (64 kDa) recognized by mAb Tugh4. However, these cells constitutively transcribe the corresponding gamma(c) gene. This apparent paradox was solved by the demonstration that polyclonal anti-gamma(c) Abs detected endoglycosidase-H-sensitive immature forms of gamma(c) (54-58 kDa) expressed by quiescent CD4 T lymphocytes and Jurkat cells. Immature gamma(c) is characterized as an intracellular component localized in the endoplasmic reticulum. Pulse-chase analysis shows that the immature gamma(c) is rapidly degraded after synthesis. After activation of CD4 T lymphocytes, and as seen in the CD4 T cell line Kit 225, the endoglycosidase-H-resistant mature form of gamma(c) is detectable at the cell surface and in the endosomal compartment. For the first time, our results demonstrate that a cytokine receptor chain may be constitutively produced as an immature form. Furthermore, this supports the notion that expression of the functional form of gamma(c) may require intracellular interactions with lineage- or subset-specific molecular partners. PMID- 11418670 TI - A distinct pathway of cell-mediated apoptosis initiated by granulysin. AB - Granulysin is an antimicrobial and tumoricidal molecule expressed in granules of CTL and NK cells. In this study, we show that granulysin damages cell membranes based upon negative charge, disrupts the transmembrane potential (Deltapsi) in mitochondria, and causes release of cytochrome c. Granulysin-induced apoptosis is blocked in cells overexpressing Bcl-2. Despite the release of cytochrome c, procaspase 9 is not processed. Nevertheless, activation of caspase 3 is observed in granulysin-treated cells, suggesting that granulysin activates a novel pathway of CTL- and NK cell-mediated death distinct from granzyme- and death receptor induced apoptosis. PMID- 11418671 TI - Protection against murine listeriosis by oral vaccination with recombinant Salmonella expressing hybrid Yersinia type III proteins. AB - In the present study, we have investigated the possibility to engage the Yersinia outer protein E (YopE) as a carrier molecule for heterologous Ag delivery by the type III secretion system of Salmonella typhimurium. Defined secretion and translocation domains of YopE were fused to the immunodominant T cell Ags listeriolysin O and p60 of Listeria monocytogenes. In vitro experiments showed that S. typhimurium allows secretion and translocation of large hybrid YopE proteins in a type III-dependent fashion. Translocation and cytosolic delivery of these chimeric proteins into host cells, but not secretion into endosomal compartments, led to efficient MHC class I-restricted Ag presentation of listerial nonamer peptides. Mice orally vaccinated with a single dose of attenuated S. typhimurium expressing translocated hybrid YopE proteins revealed high numbers of IFN-gamma-producing cells reactive with listeriolysin O 91-99 or p60 217-225, respectively. This CD8 T cell response protected mice against a challenge with L. monocytogenes. In conclusion, these findings suggest that YopE is a versatile carrier molecule for type III-mediated foreign Ag delivery by Salmonella vaccine strains. PMID- 11418672 TI - IL-8 production in human lung fibroblasts and epithelial cells activated by the Pseudomonas autoinducer N-3-oxododecanoyl homoserine lactone is transcriptionally regulated by NF-kappa B and activator protein-2. AB - The destructive pulmonary inflammation associated with Pseudomonas aeruginosa colonization is caused, in part, by the production of the chemokine IL-8, which recruits neutrophils into the lung. The Pseudomonas autoinducer, N-3 oxododecanoyl homoserine lactone (3-O-C12-HSL), is a small lipid-soluble molecule that is essential in the regulation of many P. aeruginosa virulence factors, but little is known about how it affects eukaryotic cells. In this report we demonstrate that 3-O-C12-HSL is a potent stimulator of both IL-8 mRNA and protein from human fibroblasts and epithelial cells in vitro. The IL-8 produced from these 3-O-C12-HSL-stimulated cells was found to be functionally active by inducing the chemotaxis of neutrophils. To determine a mechanism for this IL-8 induction, deletion constructs of the IL-8 promoter were examined. It was found that the DNA region between nucleotides -1481 and -546 and the transcription factor NF-kappaB were essential for the maximal induction of IL-8 by 3-O-C12-HSL. This was confirmed by EMSAs, where 3-O-C12-HSL induced a shift with both AP-2 and NF-kappaB consensus DNA. The activation of NF-kappaB and subsequent production of IL-8 were found to be regulated by a mitogen-activated protein kinase pathway. These findings support the concept that the severe lung damage that accompanies P. aeruginosa infections is caused by an exuberant neutrophil response stimulated by 3-O-C12-HSL-induced IL-8. Understanding the mechanisms of 3-O-C12-HSL activation of lung structural cells may provide a means to help control lung damage during infections with P. aeruginosa. PMID- 11418673 TI - Enhanced protection against fatal mycobacterial infection in SCID beige mice by reshaping innate immunity with IFN-gamma transgene. AB - Humans with immune-compromised conditions such as SCID are unable to control infection caused by normally nonpathogenic intracellular pathogens such as Mycobacterium bovis bacillus Calmette-Guerin. We found that SCID beige mice lacking both lymphocytes and NK cells had functionally normal lung macrophages and yet a selectively impaired response of type 1 cytokines IFN-gamma and IL-12, but not TNF-alpha, during M. bovis bacillus Calmette-Guerin infection. These mice succumbed to such infection. A repeated lung gene transfer strategy was designed to reconstitute IFN-gamma in the lung, which allowed investigation of whether adequate activation of innate macrophages could enhance host defense in the complete absence of lymphocytes. IFN-gamma transgene-based treatment was initiated 10 days after the establishment of mycobacterial infection and led to increased levels of both IFN-gamma and IL-12, but not TNF-alpha, in the lung. Lung macrophages were activated to express increased MHC molecules, type 1 cytokines and NO, and increased phagocytic and mycobactericidal activities. Activation of innate immunity markedly inhibited otherwise uncontrollable growth of mycobacteria and prolonged the survival of infected SCID hosts. Thus, our study proposes a cytokine transgene-based therapeutic modality to enhance host defense in immune-compromised hosts against intracellular bacterial infection, and suggests a central effector activity played by IFN-gamma-activated macrophages in antimycobacterial cell-mediated immunity. PMID- 11418674 TI - Dendritic cells recruitment and in vivo priming of CD8+ CTL induced by a single topical or transepithelial immunization via the buccal mucosa with measles virus nucleoprotein. AB - The buccal mucosa, a prototype of pluristratified mucosal epithelia, contains a network of directly accessible class II(+) epithelial dendritic cells (DC), similar to skin Langerhans cells. We showed that a single buccal immunization with measles virus nucleoprotein (NP), by either topical application onto or intradermal injection in the buccal mucosa, induced in vivo priming of protective class I-restricted specific CD8(+) CTL. Both routes of immunization with NP induced a rapid recruitment of DC into the mucosa, which peaked at 2 h and decreased by 24 h. Treatment of mice with Flt3 ligand resulted in an increased number of DC in the buccal mucosa and enhanced the frequency of IFN-gamma producing NP-specific effectors and the NP-specific CTL response generated after buccal immunization with NP. Finally, NP-pulsed bone marrow-derived DC induced NP specific IFN-gamma-producing cells upon adoptive transfer to naive mice. These data demonstrate that a viral protein delivered to DC of the buccal mucosa induces in vivo priming of protective anti-viral CD8(+) CTL. PMID- 11418675 TI - IFN-gamma-independent autocrine cytokine regulatory mechanism in reprogramming of macrophage responses to bacterial lipopolysaccharide. AB - Macrophages are now well recognized to have a critical role in both innate and acquired immunity. The sentinel macrophage function is highly regulated and serves to allow for intrinsic plasticity of the innate immune responses to potential environmental signals. However, the mechanisms underlying the dynamic properties of the cellular arm of innate immunity are poorly understood. Therefore, we have conducted a series of in vitro studies to evaluate the contribution of immunoregulatory cytokines, such as IFN-gamma, IL-10, and IL-12, in modulation of macrophage responses. We found that macrophages from IFN-gamma knockout (IFN-gamma(-/-)) mice exhibit only marginal LPS-induced TNF-alpha, IL 12, and NO responses, all of which can be fully restored in the presence of rIFN gamma. Pretreatment with substimulatory LPS concentrations led to reprogramming of IFN-gamma(-/-) macrophage responses in a dose-dependent manner that manifested by an increased TNF-alpha and IL-12, but not NO, production upon the subsequent LPS challenge. These reprogramming effects were substantially attenuated and profoundly enhanced in macrophages from IL-12(-/-) and IL-10(-/-) mice, respectively, as compared with those modulated in macrophages from the congenic wild-type mice. LPS-dependent reprogramming was also fully reproduced in macrophages isolated from SCID mice after immunodepletion of NK cells. Our data strongly imply that cytokine (TNF-alpha and IL-12), but not NO, responses in macrophages may, at least in part, be governed by an autocrine IFN-gamma independent regulatory mechanism reciprocally controlled by IL-10 and IL-12. This mechanism may serve as an alternative/coherent pathway to the canonical IFN-gamma dependent induction of antimicrobial and tumoricidal activity in macrophages. PMID- 11418676 TI - Cytokine-responsive gene-2/IFN-inducible protein-10 expression in multiple models of liver and bile duct injury suggests a role in tissue regeneration. AB - IFN-inducible protein-10 (IP-10/CXCL10) is a CXC chemokine that targets both T cells and NK cells. Elevation of IP-10 expression has been demonstrated in a number of human diseases, including chronic cirrhosis and biliary atresia. Cytokine-responsive gene-2 (Crg-2), the murine ortholog of IP-10, was induced following CCl(4) treatment of the hepatocyte-like cell line AML-12. Crg-2 expression was noted in vivo in multiple models of hepatic and bile duct injury, including bile duct ligation and CCl(4), D-galactosamine, and methylene dianiline toxic liver injuries. Induction of Crg-2 was also examined following two-thirds hepatectomy, a model that minimally injures the remaining liver, but that requires a large hepatic regenerative response. Crg-2 was induced in a biphasic fashion after two-thirds hepatectomy, preceding each known peak of hepatocyte DNA synthesis. Induction of Crg-2 was also observed in the kidney, gut, thymus, and spleen within 1 h of two-thirds hepatectomy. Characteristic of an immediate early gene, pretreatment of mice with the protein synthesis inhibitor cycloheximide before either two-thirds hepatectomy or CCl(4) injection led to Crg-2 superinduction. rIP-10 was demonstrated to have hepatocyte growth factor-inducing activity in vitro, but alone had no direct mitogenic effect on hepatocytes. Our data demonstrate that induction of Crg-2 occurs in several distinct models of liver injury and regeneration, and suggest a role for CRG-2/IP-10 in these processes. PMID- 11418677 TI - Impaired mucosal immunity in L-selectin-deficient mice orally immunized with a Salmonella vaccine vector. AB - Lymphocyte trafficking in the gastrointestinal tract is primarily mediated by interactions with the mucosal addressin cell adhesion molecule 1 and its lymphocyte ligand, alpha(4)beta(7), and partly by L-selectin (L-Sel) interactions with peripheral node addressin coexpressed on some mucosal addressin cell adhesion molecule 1. We inquired whether intestinal responses in mice lacking L Sel would be enhanced. L-Sel-deficient (L-Sel(-/-)) mice were orally immunized with either Salmonella vaccine vector or Salmonella vector-expressing colonization factor Ag I (CFA/I) from enterotoxigenic Escherichia coli. In L-Sel( /-) mice, mucosal IgA anti-CFA/I fimbrial responses were greatly reduced, and systemic IgG2a anti-CFA/I fimbrial responses were 26-fold greater compared with C57BL/6 (L-Sel(+/+)) mice. L-Sel(-/-) Peyer's patch (PP) CD4(+) Th cells revealed IFN-gamma-dominated responses and an unprecedented absence of IL-4, whereas the expected mixed Th cell phenotype developed in L-Sel(+/+) mice. PP CD4(+) Th cell anti-Salmonella responses were nearly nonexistent in L-Sel(-/-) mice immunized with either Salmonella vaccine. Splenic CD4(+) Th cell anti-Salmonella responses were reduced but did show cytokine production in Ag restimulation assays. Increased colonization of PP and spleen was noted only with the Salmonella vector in L-Sel(-/-) mice, resulting in increased splenomegaly, suggesting that the Salmonella-CFA/I vaccine was not as infectious or that the presence of the fimbriae improved clearance, possibly because of reduced neutrophil recruitment. However, sufficient anti-Salmonella immunity was induced, because Salmonella vector-immunized L-Sel(-/-) mice showed complete protection against wild-type Salmonella challenge, unlike L-Sel(+/+) mice. This evidence shows that L-Sel is important for development of mucosal immunity, and absence of L-Sel is protective against salmonellosis. PMID- 11418678 TI - Activation of Toll-like receptor-2 by glycosylphosphatidylinositol anchors from a protozoan parasite. AB - Glycosylphosphatidylinositol (GPI) anchors and glycoinositolphospholipids (GIPLs) from parasitic protozoa have been shown to exert a wide variety of effects on cells of the host innate immune system. However, the receptor(s) that are triggered by these protozoan glycolipids has not been identified. Here we present evidence that Trypanosoma cruzi-derived GPI anchors and GIPLs trigger CD25 expression on Chinese hamster ovary-K1 cells transfected with CD14 and Toll-like receptor-2 (TLR-2), but not wild-type (TLR-2-deficient) Chinese hamster ovary cells. The protozoan-derived GPI anchors and GIPLs containing alkylacylglycerol and saturated fatty acid chains or ceramide were found to be active in a concentration range of 100 nM to 1 microM. More importantly, the GPI anchors purified from T. cruzi trypomastigotes, which contain a longer glycan core and unsaturated fatty acids in the sn-2 position of the alkylacylglycerolipid component, triggered TLR-2 at subnanomolar concentrations. We performed experiments with macrophages from TLR-2 knockout and TLR-4 knockout mice, and found that TLR-2 expression appears to be essential for induction of IL-12, TNF alpha, and NO by GPI anchors derived from T. cruzi trypomastigotes. Thus, highly purified GPI anchors from T. cruzi parasites are potent activators of TLR-2 from both mouse and human origin. The activation of TLR-2 may initiate host innate defense mechanisms and inflammatory response during protozoan infection, and may provide new strategies for immune intervention during protozoan infections. PMID- 11418679 TI - Role of effector cell-derived IL-4, IL-5, and perforin in early and late stages of type 2 CD8 effector cell-mediated tumor rejection. AB - Type 2 CD8 T cells (Tc2) secrete IL-4 and IL-5 and display perforin-dependent cytolysis in vitro. Using an OVA-transfected B16-melanoma model, we show that tumor-reactive Tc2 effector cells accumulated at the tumor site and induced tumor regression that enhanced survival in mice with pulmonary tumors. Transfer of perforin-deficient Tc2 cells generated from perforin gene knockout mice showed no differences in therapeutic efficiency when compared with wild-type Tc2 cells. In contrast, Tc2 cells derived from select cytokine gene-deficient mice showed that therapeutic effects were dependent on effector cell-derived IL-4 and IL-5 that led to a local elevation in lung-derived chemoattractants and accumulation of activated host-derived CD8/CD44(high), CD4/CD44(high), and OVA-specific tetramer positive CD8 cells in vivo. Host-derived T and non-T immune cells increased in the lung over time and correlated with an elevated production of type 1-related chemokines. Conversely, donor Tc2 cell numbers markedly diminished at later times, suggesting that prolonged therapeutic responses were due to host-derived mechanisms. Moreover, type 1 host responses were detectable with increased levels of IFN-gamma production by lung-derived CD4 and CD8 T cells from surviving Tc2 treated mice. Transfer of Tc2 cells into IFN-gamma-deficient tumor-bearing mice was markedly less effective then into wild-type mice, suggesting that host derived IFN-gamma-dependent mechanisms play a role in Tc2-mediated antitumor responses. PMID- 11418680 TI - Induction of apoptosis in human T cells by Actinobacillus actinomycetemcomitans cytolethal distending toxin is a consequence of G2 arrest of the cell cycle. AB - We have previously shown that Actinobacillus actinomycetemcomitans produces an immunosuppressive factor that is encoded by the cdtB gene, which is homologous to a family of cytolethal distending toxins (Cdt) expressed by several Gram-negative bacteria. Moreover, we have shown that CdtB impairs lymphocyte function by inducing G(2) arrest of the cell cycle. We now report that both CdtB as well as an extract prepared from an Escherichia coli strain that expresses all three of the A. actinomycetemcomitans cdt genes (rCdtABC) induce apoptosis. Pretreatment of lymphocytes with either CdtB or rCdtABC leads to DNA fragmentation in activated lymphocytes at 72 and 96 h. No DNA fragmentation was induced in nonactivated cells. Flow cytometric analysis of the Cdt-treated lymphocytes demonstrates a reduction in cell size and an increase in nuclear condensation. Mitochondrial function was also perturbed in cells pretreated with either CdtB or rCdtABC. An increase in the expression of the mitochondria Ag, Apo 2.7, was observed along with evidence of the development of a mitochondrial permeability transition state; this includes a decrease in the transmembrane potential and elevated generation of reactive oxygen species. Activation of the caspase cascade, which is an important biochemical feature of the apoptotic process, was also observed in Cdt-treated lymphocytes. Overexpression of the bcl-2 gene in the human B lymphoblastoid cell line, JY, led to a decrease in Cdt-induced apoptosis. Interestingly, Bcl-2 overexpression did not block Cdt-induced G(2) arrest. The implications of our results with respect to the immunosuppressive functions of Cdt proteins are discussed. PMID- 11418681 TI - Lacto-N-fucopentaose III found on Schistosoma mansoni egg antigens functions as adjuvant for proteins by inducing Th2-type response. AB - We have recently demonstrated that induction of Th2 responses by Schistosoma mansoni egg Ag is largely due to carbohydrates on the Ag functioning as adjuvants. Lacto-N-fucopentaose III (LNFPIII), a polylactosamine sugar, is the predominant carbohydrate found in S. mansoni egg Ag. Therefore, using neoglycoprotein, we investigated whether LNFPIII induces in vivo Th2 response and functions as an adjuvant. Following intranasal immunization with LNFPIII linked to human serum albumin (HSA) (HSA-LNFPIII), BALB/c mice mounted a strong Th2 response and produced significantly higher levels of total IgE as well as HSA specific IgG, IgG1, and IgE. HSA-LNFPIII was over 1000-fold more potent in inducing Ab production as compared with HSA alone. Although LNFPIII itself did not function as an epitope for either IgG or IgE, its conjugation with protein was essential for the adjuvant activity. Moreover, fucose residue on LNFPIII was crucial for induction of Ab production. Nasal lymphocytes from mice immunized with HSA-LNFPIII produced IL-4, IL-5, and IL-10, but not IFN-gamma following in vitro stimulation with HSA or HSA-LNFPIII. In addition, these activated nasal lymphocytes also showed a significant increase of B7-2 expression on B220 positive cells. Furthermore, not only intranasal but also both i.p. and s.c. immunization with HSA-LNFPIII induced significant production of HSA-specific Abs compared with the immunization with HSA alone, suggesting that the activity of LNFPIII was not restricted on particular route of immunization. These results demonstrate that Lewis type carbohydrate LNFPIII can function as an adjuvant by their ability to induce a Th2 response. PMID- 11418682 TI - Serum proteins modified by neutrophil-derived oxidants as mediators of neutrophil stimulation. AB - Reactive oxygen intermediates (ROI) released during inflammation may act as important mediators of neutrophil effector functions. The objective of this investigation was to evaluate the influence of ROI generation on neutrophil adhesion molecule regulation and degranulation. Induction of the neutrophil oxidative burst via Fcgamma receptor cross-linking was accompanied by up regulation of neutrophil surface CD11b, CD35, and CD66b only in the presence of selected serum proteins, such as purified human C4, C5, or human serum albumin (HSA). Scavenging of ROI attenuated protein-dependent receptor regulations. Moreover, exogenous hydrogen peroxide was effective to increase neutrophil CD11b expression in a protein-dependent way. HSA exposed to neutrophil-derived ROI displayed signs of oxidative modification in terms of carbonyl formation. Such modified HSA transferred to resting neutrophils bound readily to the cell surface and effected receptor modulation as well as cellular spreading. In contrast, neither native HSA nor HSA protected against oxidation by the tocopherol analog Trolox exhibited agonistic properties. In conclusion, we demonstrate that neutrophil-derived ROI modify selected serum proteins, which, in turn, act as proinflammatory mediators of neutrophil stimulation. PMID- 11418683 TI - Role of mitogen-activated protein kinase-mediated cytosolic phospholipase A2 activation in arachidonic acid metabolism in human eosinophils. AB - The objective of this investigation was to determine the role of secretory and cytosolic isoforms of phospholipase A(2) (PLA(2)) in the induction of arachidonic acid (AA) and leukotriene synthesis in human eosinophils and the mechanism of PLA(2) activation by mitogen-activated protein kinase (MAPK) isoforms in this process. Pharmacological activation of eosinophils with fMLP caused increased AA release in a concentration (EC(50) = 8.5 nM)- and time-dependent (t(1/2) = 3.5 min) manner. Both fMLP-induced AA release and leukotriene C(4) (LTC(4)) secretion were inhibited concentration dependently by arachidonic trifluoromethyl ketone, a cytosolic PLA(2) (cPLA(2)) inhibitor; however, inhibition of neither the 14-kDa secretory phospholipase A(2) by 3-(3-acetamide-1-benzyl-2-ethylindolyl-5 oxy)propanephosphonic acid nor cytosolic Ca(2+)-independent phospholipase A(2) inhibition by bromoenol lactone blocked hydrolysis of AA or subsequent leukotriene synthesis. Pretreatment of eosinophils with a mitogen-activated protein/extracellular signal-regulated protein kinase (ERK) kinase inhibitor, U0126, or a p38 MAPK inhibitor, SB203580, suppressed both AA production and LTC(4) release. fMLP induced phosphorylation of MAPK isoforms, ERK1/2 and p38, which were evident after 30 s, maximal at 1-5 min, and declined thereafter. fMLP stimulation also increased cPLA(2) activity in eosinophils, which was inhibited completely by 30 microM arachidonic trifluoromethyl ketone. Preincubation of eosinophils with U0126 or SB203580 blocked fMLP-enhanced cPLA(2) activity. Furthermore, inhibition of Ras, an upstream GTP-binding protein of ERK, also suppressed fMLP-stimulated AA release. These findings demonstrate that cPLA(2) activation causes AA hydrolysis and LTC(4) secretion. We also find that cPLA(2) activation caused by fMLP occurs subsequent to and is dependent upon ERK1/2 and p38 MAPK activation. Other PLA(2) isoforms native to human eosinophils possess no significant activity in the stimulated production of AA or LTC(4). PMID- 11418684 TI - Microsomal prostaglandin E synthase is regulated by proinflammatory cytokines and glucocorticoids in primary rheumatoid synovial cells. AB - The selective induction of PGE(2) synthesis in inflammation suggests that a PGE synthase may be linked to an inducible pathway for PG synthesis. We examined the expression of the recently cloned inducible microsomal PGE synthase (mPGES) in synoviocytes from patients with rheumatoid arthritis, its modulation by cytokines and dexamethasone, and its linkage to the inducible cyclooxygenase-2. Northern blot analysis showed that IL-1beta or TNF-alpha treatment induces mPGES mRNA from very low levels at baseline to maximum levels at 24 h. IL-1beta-induced mPGES mRNA was inhibited by dexamethasone in a dose-dependent fashion. Western blot analysis demonstrated that mPGES protein was induced by IL-1beta, and maximum expression was sustained for up to 72 h. There was a coordinated up-regulation of cyclooxygenase-2 protein, although peak expression was earlier. Differential Western blot analysis of the microsomal and the cytosolic fractions revealed that the induced expression of mPGES protein was limited to the microsomal fraction. The detected mPGES protein was catalytically functional as indicated by a 3-fold increase of PGES activity in synoviocytes following treatment with IL-1beta; this increased synthase activity was limited to the microsomal fraction. In summary, these data demonstrate an induction of mPGES in rheumatoid synoviocytes by proinflammatory cytokines. This novel pathway may be a target for therapeutic intervention for patients with arthritis. PMID- 11418685 TI - Gi-mediated activation of the Syk kinase by the receptor mimetic basic secretagogues of mast cells: role in mediating arachidonic acid/metabolites release. AB - Syk kinase is essential for FcepsilonRI-mediated signaling and release of inflammatory mediators from mast cells. We now show that activation of rat peritoneal mast cells by the nonimmunological, G(i)-mediated pathway also results in the activation of Syk. We show that compound 48/80 (c48/80), a receptor analogue that activates directly G proteins, activates Syk in a pertussis toxin sensitive fashion. We further show that Syk activation by c48/80 is blocked by the protein kinase C inhibitor GF109203X, by the phosphatidylinositol 3-kinase inhibitors, wortmannin and LY294002, by EGTA, and by the selective src-like kinase inhibitor PP1. These results suggest that in the nonimmunological, G(i) mediated pathway, Syk is located downstream from phospholipase C and phosphatidylinositol 3-kinase. However, in common with the FcepsilonRI-mediated pathway, activation of Syk by c48/80 is dependent on a src-like protein tyrosine kinase. Finally, we show that in the nonimmunological pathway, Syk plays a central role in the release of arachidonic acid/eicosanoid metabolites, but not in the release of prestored mediators such as histamine. PMID- 11418686 TI - Diphosphoryl lipid A from Rhodobacter sphaeroides blocks the binding and internalization of lipopolysaccharide in RAW 264.7 cells. AB - Diphosphoryl lipid A derived from the nontoxic LPS of Rhodobacter sphaeroides (RsDPLA) has been shown to be a powerful LPS antagonist in both human and murine cell lines. In addition, RsDPLA also can protect mice against the lethal effects of toxic LPS. In this study, we complexed both the deep rough LPS from Escherichia coli D31 m4 (ReLPS) and RsDPLA with 5- and 30-nm colloidal gold and compared their binding to the RAW 264.7 cell line by electron microscopy. Both ReLPS and RsDPLA bound to the cells with the following observations. First, binding studies revealed that pretreatment with RsDPLA completely blocked the binding and thus internalization of ReLPS-gold conjugates to these cells at both 37 degrees C and 4 degrees C. Second, ReLPS was internalized via micropinocytosis (noncoated plasma membrane invaginations) involving formation of caveolae-like structures and leading to the formation of micropinocytotic vesicles, macropinocytosis (or phagocytosis), formation of clathrin-coated pits (receptor mediated), and penetration through plasma membrane into cytoplasm. Third, in contrast, RsDPLA was internalized predominantly via macropinocytosis. These studies show for the first time that RsDPLA blocks the binding and thus internalization of LPS as observed by scanning and transmission electron microscopy. PMID- 11418687 TI - Gammalinolenic acid, an unsaturated fatty acid with anti-inflammatory properties, blocks amplification of IL-1 beta production by human monocytes. AB - Administration of gammalinolenic acid (GLA), an unsaturated fatty acid, reduces joint inflammation in patients with rheumatoid arthritis. Addition of GLA in vitro suppresses release of IL-1beta from human monocytes stimulated with LPS. LPS-induced IL-1beta release is followed by IL-1-induced IL-1beta release, an amplification process termed autoinduction. We show here with peripheral blood monocytes from normal volunteers and from patients with rheumatoid arthritis by using IL-1R antagonist to block autoinduction and IL-1alpha stimulation to simulate autoinduction that approximately 40% of IL-1beta released from LPS stimulated cells is attributable to autoinduction and that GLA reduces autoinduction of IL-1beta while leaving the initial IL-1beta response to LPS intact. Experiments with cells in which transcription and protein synthesis were blocked suggest that GLA induces a protein that reduces pro-IL-1beta mRNA stability. IL-1beta is important to host defense, but the amplification mechanism may be excessive in genetically predisposed patients. Thus, reduction of IL-1beta autoinduction may be protective in some patients with endotoxic shock and with diseases characterized by chronic inflammation. PMID- 11418688 TI - Both E6 and E7 oncoproteins of human papillomavirus 16 inhibit IL-18-induced IFN gamma production in human peripheral blood mononuclear and NK cells. AB - Cervical carcinoma is the predominant cancer among malignancies in women throughout the world, and human papillomavirus (HPV) 16 is the most common agent linked to human cervical carcinoma. The present study was performed to investigate the mechanisms of immune escape in HPV-induced cervical cancer cells. The presence of HPV oncoproteins E6 and E7 in the extracellular fluids of HPV containing cervical cancer cell lines SiHa and CaSki was demonstrated by ELISA. The effect of HPV 16 oncoproteins E6 and E7 on the production of IFN-gamma by IL 18 was assessed. E6 and E7 proteins reduced IL-18-induced IFN-gamma production in both primary PBMCs and the NK0 cell line. FACS analysis revealed that the viral oncoproteins reduced the binding of IL-18 to its cellular surface receptors on NK0 cells, whereas there was no effect of oncoproteins on IL-1 binding to its surface IL-1 receptors on D10S, a subclone of the murine Th cell D10.G4.1. In vitro pull-down assays also revealed that the viral oncoproteins and IL-18 bound to IL-18R alpha-chain competitively. These results suggest that the extracellular HPV 16 E6 and E7 proteins may inhibit IL-18-induced IFN-gamma production locally in HPV lesions through inhibition of IL-18 binding to its alpha-chain receptor. Down-modulation of IL-18-induced immune responses by HPV oncoproteins may contribute to viral pathogenesis or carcinogenesis. PMID- 11418689 TI - Kaposi's sarcoma-associated herpesvirus G protein-coupled receptor constitutively activates NF-kappa B and induces proinflammatory cytokine and chemokine production via a C-terminal signaling determinant. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is believed to be the causative agent of Kaposi's sarcoma (KS), a multicentric growth factor-dependent tumor common in AIDS patients characterized histopathologically by spindle cell proliferation, angiogenesis, and leukocyte infiltration. Recently, open reading frame 74 of KSHV has been implicated as a major viral determinant of KS. Open reading frame 74 encodes KSHV G protein-coupled receptor (GPCR), a constitutively active chemokine receptor that directly transforms NIH 3T3 cells in vitro and induces multifocal KS-like lesions in KSHV-GPCR-transgenic mice. Interestingly, receptor-positive cells are very rare in lesions from these mice, implicating an indirect mechanism of tumorigenesis. In this regard, here we report that expression of KSHV-GPCR in transfected epithelial, monocytic, and T cell lines induced constitutive activation of the immunoregulatory transcription factors AP 1 and NF-kappaB. This was associated with constitutive induction of the proinflammatory NF-kappaB-dependent cytokines IL-1beta, IL-6, and TNF-alpha, and chemokines monocyte chemoattractant protein-1 and IL-8, as well as the AP-1 dependent basic fibroblast growth factor. In addition, IL-2 and IL-4 production was induced in transfected Jurkat T cells. Truncation of the final five amino acids in the cytoplasmic tail of KSHV-GPCR caused complete loss of its transforming and NF-kappaB-inducing activities, without affecting receptor expression or ligand binding. These data suggest that KS results in part from KSHV-GPCR induction of proinflammatory cytokine and growth factor gene expression, mediated by a signaling determinant within the last five amino acids of the C terminus, a domain that is also critical for direct cell transformation. PMID- 11418690 TI - Dissection of the intracellular pathways in hepatocytes suggests a role for Jun kinase and IFN regulatory factor-1 in Con A-induced liver failure. AB - Con A administration results in dose-dependent immune-mediated liver injury. Cytokines are important to determine the outcome of liver failure in this model, and especially TNF-alpha and IFN-gamma directly contribute to hepatocyte damage. The intracellular pathways of these two cytokines, which eventually result in tissue destruction, are not well defined. Here we used anti-IFN-gamma Abs and adenoviral vectors that express molecules inhibiting distinct TNF-alpha-dependent pathways in hepatocytes to better understand the relevance of specific intracellular signaling cascades for Con A-induced liver failure. We show that activation of TNF-alpha- and IFN-gamma-dependent intracellular pathways occurs prior to the influx of immune-activated cells into the liver and that anti-TNF alpha and anti-IFN-gamma neutralizing Abs cannot block infiltration of these cells. Blocking experiments with Abs and adenoviral vectors showed that NF-kappaB activation and the Fas-associated death domain protein/caspase 8 cascade in hepatocytes during Con A-induced liver failure have no impact on tissue injury. Additionally, STAT1 activation alone after Con A injection in liver cells does not result in liver damage. In contrast, IFN-gamma-dependent expression of IFN regulatory factor-1 and TNF-alpha-dependent activation of c-Jun N-terminal kinase in liver cells correlates with liver cell damage after Con A injection. Therefore, our experiments indicate that 11418690 PMID- 11418691 TI - Stem cell factor-induced leukotriene B4 production cooperates with eotaxin to mediate the recruitment of eosinophils during allergic pleurisy in mice. AB - The understanding of the mechanisms underlying eosinophil recruitment in vivo may aid in the development of novel strategies for the treatment of allergic disorders. In this study, we investigated the role of chemokines in the cascade of events leading to eosinophil recruitment in a stem cell factor (SCF)- and leukotriene B(4) (LTB(4))-dependent allergic pleurisy model in mice. The intrapleural administration of the eosinophil-active chemokines eotaxin, RANTES, and macrophage-inflammatory protein 1alpha (MIP-1alpha) induced a time- and dose dependent eosinophil recruitment. Pretreatment with anti-eotaxin, but not anti RANTES or anti-MIP-1alpha, blocked the recruitment of eosinophils following Ag challenge of sensitized animals, and significant eotaxin immunoreactivity was detected in the pleural cavity of these animals. Similarly, only the anti-eotaxin inhibited the eosinophil recruitment induced by injection of SCF in naive animals. However, blockade of SCF did not inhibit the release of eotaxin after Ag challenge of sensitized mice. Akin to its effects on SCF and in the allergic reaction, eotaxin-induced eosinophil recruitment was blocked by the LTB(4) receptor antagonist CP105696. Nevertheless, SCF, but not eotaxin, appeared to regulate the endogenous release of LTB(4) after Ag challenge. Finally, we show that low doses of eotaxin synergized with LTB(4) to induce eosinophil recruitment in the pleural cavity. Overall, the present results show that eotaxin and SCF induced LTB(4) cooperate to induce eosinophil recruitment into sites of allergic inflammation. Cooperation between inflammatory mediators must be an important phenomenon in vivo, explaining both the ability of lower concentrations of mediators to induce a full-blown functional response and the effectiveness of different strategies at inhibiting these responses. PMID- 11418692 TI - Critical role for IL-4 in the development of transplant arteriosclerosis in the absence of CD40-CD154 costimulation. AB - Blockade of the CD40-CD154 pathway can inhibit CD4(+) T cell activation but is unable to prevent immune responses mediated by CD8(+) T cells. However, even in the absence of CD8(+) T cells, inhibition of the CD40-CD154 pathway is insufficient to prevent the development of transplant arteriosclerosis. This study investigated the mechanisms of transplant arteriosclerosis in the absence of the CD40 pathway. C57BL/6 CD40(-/-) (H2(b)) recipients were transplanted with MHC-mismatched BALB/c (H2(d)) aortas. Transplant arteriosclerosis was evident in both CD40(-/-) and CD40(+/-) mice (intimal proliferation was 59 +/- 5% for CD40( /-) mice vs 58 +/- 4% for CD40(+/-) mice) in the presence or absence of CD8(+) T cells (intimal proliferation was 46 +/- 7% for CD40(-/-) anti-CD8-treated mice vs 50 +/- 10% for CD40(+/-) anti-CD8-treated mice), confirming that CD8(+) T cells are not essential effector cells for the development of this disease. In CD40(-/ ) recipients depleted of CD8(+) T cells, the number of eosinophils infiltrating the graft was markedly increased (109 +/- 24 eosinophils/grid for CD40(-/-) anti CD8-treated mice vs 28 +/- 7 for CD40(+/-) anti-CD8-treated mice). The increased presence of eosinophils correlated with augmented intragraft production of IL-4. To test the hypothesis that IL-4 was responsible for the intimal proliferation, CD8 T cell-depleted CD40(-/-) recipients were treated with anti-IL-4 mAb. This resulted in significantly reduced eosinophil infiltration into the graft (12 +/- 5 eosinophils/grid for CD40(-/-) anti-CD8(+), anti-IL-4-treated mice vs 109 +/- 24 for CD40(-/-) anti-CD8-treated mice), intragraft eotaxin, CCR3 mRNA production, and the level of intimal proliferation (18 +/- 5% for CD40(-/-) anti CD8(+)-, anti-IL-4-treated mice vs 46 +/- 7% for CD40(-/-) anti-CD8-treated mice). In conclusion, elevated intragraft IL-4 production results in an eosinophil infiltrate and is an important mechanism for CD8(+) T cell-independent transplant arteriosclerosis in the absence of CD40-CD154 costimulation. PMID- 11418693 TI - Estrogen treatment down-regulates TNF-alpha production and reduces the severity of experimental autoimmune encephalomyelitis in cytokine knockout mice. AB - A shift toward Th2 cytokine production has been demonstrated during pregnancy and high dose estrogen therapy and is thought to be the primary mechanism by which estrogen suppresses the development of experimental autoimmune encephalomyelitis. However, low dose estrogen treatment is equally protective in the absence of a significant shift in cytokine production. In this study cytokine-deficient mice were treated with estrogen to determine whether a shift in Th2 cytokine production was required for the protective effects of hormone therapy. Estrogen effectively suppressed the development of experimental autoimmune encephalomyelitis in IL-4 and IL-10 knockout mice and in wild type littermate mice with a similar potency of protection. Significant disease suppression was also seen in IFN-gamma-deficient mice. The decrease in disease severity was accompanied by a concomitant reduction in the number of proinflammatory cytokine- and chemokine-producing cells in the CNS. Although there was no apparent increase in compensatory Th2 cytokine production in cytokine-deficient mice, there was a profound decrease in the frequency of TNF-alpha-producing cells in the CNS and the periphery. Therefore, we propose that one mechanism by which estrogen protects females from the development of cell-mediated autoimmunity is through a hormone-dependent regulation of TNF-alpha production. PMID- 11418694 TI - Suppression of IL-2-induced T cell proliferation and phosphorylation of STAT3 and STAT5 by tumor-derived TGF beta is reversed by IL-15. AB - IL-2 responses are susceptible to suppression by TGFbeta, a cytokine widely implicated in suppression of inflammatory responses and secreted by many different tumor cell types. There have been conflicting reports regarding inhibition of IL-2-induced STAT3 and STAT5 phosphorylation by TGFbeta and subsequent suppression of immune responses. Using TGFbeta-producing multiple myeloma tumor cells we demonstrate that tumor-derived TGFbeta can block IL-2 induced proliferation and STAT3 and STAT5 phosphorylation in T cells. High affinity IL-2R expression was required for the suppression of IL-2 responses as a novel CD25(-) T cell line proliferated and phosphorylated STAT3 when cultured with tumor cells or rTGFbeta1. Activating T cells with IL-15, which does not use the high affinity IL-2R, completely restored the ability of T cells to phosphorylate STAT3 and STAT5 when cultured with tumor cells. IL-15-treated T cells proliferated normally when cocultured with tumor cells or rTGFbeta1, whereas IL-2 responses were consistently inhibited. Preincubation with IL-15 also restored the ability of T cells to respond to IL-2 by phosphorylating STAT3 and STAT5, and proliferating normally in the presence of tumor cells. IL-2 pretreatment did not restore T cell function. IL-15 also restored T cell responses by T cells from multiple myeloma patients, and against freshly isolated bone marrow tumor samples. Thus, activation of T cells by IL-15 renders T cells resistant to suppression by TGFbeta1-producing tumor cells and rTGFbeta1. This finding may be exploited in the design of new immunotherapy approaches that will rely on T cells avoiding tumor-induced suppression. PMID- 11418695 TI - T cell epitope mapping of the Smith antigen reveals that highly conserved Smith antigen motifs are the dominant target of T cell immunity in systemic lupus erythematosus. AB - B cell and T cell immunity to the Smith Ag (Sm) is a characteristic feature of systemic lupus erythematosus (SLE). We have shown that T cell immunity against Sm can be detected in SLE patients, and that T and B cell immunity against Sm are linked in vivo. TCR usage by Sm-reactive T cells is highly restricted and characteristic of an Ag-driven immune response. Sm is a well-characterized complex Ag consisting of proteins B1, B2, D1, D2, D3, E, F, and G. A unique feature of all Sm proteins is the presence of homologous motifs, Sm motif 1 and Sm motif 2. We used limiting dilution cloning and synthetic peptide Ags to characterize the human T cell immune response against Sm in seven SLE patients. We sought to determine the precise antigenic peptides recognized, the common features of antigenic structure recognized, and the evolution of the T cell response against Sm. We found there was a highly restricted set of Sm self peptides recognized by T cells, with three epitopes on Sm-B and two epitopes on Sm-D. We found that T cell immunity against Sm-B and Sm-D was encoded within the highly conserved Sm motif 1 and Sm motif 2, and that immunity against these epitopes appeared stable. The present study supports the concept that T cell immunity to Sm is an Ag-driven immune response directed against a highly restricted set of self-peptides, encoded within Sm motif 1 and Sm motif 2, that is shared among all Sm proteins. PMID- 11418696 TI - Therapeutic alteration of insulin-dependent diabetes mellitus progression by T cell tolerance to glutamic acid decarboxylase 65 peptides in vitro and in vivo. AB - We have reported previously that nonobese diabetic (NOD) fetal pancreas organ cultures lose the ability to produce insulin when maintained in contact with NOD fetal thymus organ cultures (FTOC). Initial studies indicated that exposure to glutamic acid decarboxylase (GAD65) peptides in utero resulted in delay or transient protection from insulin-dependent diabetes mellitus (IDDM) in NOD mice. We also found that exposure of young adult NOD mice to the same peptides could result in acceleration of the disease. To more closely examine the effects of early and late exposure to diabetogenic Ags on T cells, we applied peptides derived from GAD65 (GAD AA 246-266, 509-528, and 524-543), to our "in vitro IDDM" (ivIDDM) model. T cells derived from NOD FTOC primed during the latter stages of organ culture, when mature T cell phenotypes are present, had the ability to proliferate to GAD peptides. ivIDDM was exacerbated under these conditions, suggesting that GAD responsiveness correlates with the ivIDDM phenotype, and parallels the acceleration of IDDM we had seen in young adult NOD mice. When GAD peptides were present during the initiation of FTOC, GAD proliferative responses were inhibited, and ivIDDM was reduced. This result suggests that tolerance to GAD peptides may reduce the production of diabetogenic T cells or their capacity to respond, as suggested by the in utero therapies studied in NOD mice. PMID- 11418697 TI - A retro-inverso peptide mimic of CD28 encompassing the MYPPPY motif adopts a polyproline type II helix and inhibits encephalitogenic T cells in vitro. AB - Complete activation of T cells requires two signals: an Ag-specific signal delivered via the TCR by the peptide-MHC complex and a second costimulatory signal largely provided by B7:CD28/CTLA-4 interactions. Previous studies have shown that B7 blockade can either ameliorate experimental autoimmune encephalomyelitis by interfering with CD28 signaling or exacerbate the disease by concomitant blockade of CTLA-4 interaction. Therefore, we developed a functional CD28 mimic to selectively block B7:CD28 interactions. The design, synthesis, and structural and functional properties of the CD28 free peptide, the end group blocked CD28 peptide, and its retro-inverso isomer are shown. The synthetic T cell-costimulatory receptor peptides fold into a polyproline type II helical structure commonly seen in regions of globular proteins involved in transient protein-protein interactions. The binding determinants of CD28 can be transferred onto a short peptide mimic of its ligand-binding region. The CD28 peptide mimics effectively block the expansion of encephalitogenic T cells in vitro suggesting the potential usefulness of the peptides for the treatment of autoimmune disease conditions requiring down-regulation of T cell responses. PMID- 11418698 TI - Plasmid DNAs encoding insulin and glutamic acid decarboxylase 65 have distinct effects on the progression of autoimmune diabetes in nonobese diabetic mice. AB - We previously demonstrated that administration of plasmid DNAs (pDNAs) encoding IL-4 and a fragment of glutamic acid decarboxylase 65 (GAD65) fused to IgGFc induces GAD65-specific Th2 cells and prevents insulin-dependent diabetes mellitus (IDDM) in nonobese diabetic (NOD) mice. To assess the general applicability of pDNA vaccination to mediate Ag-specific immune deviation, we examined the immunotherapeutic efficacy of recombinants encoding murine insulin A and B chains fused to IgGFc. Insulin was chosen based on studies demonstrating that administration of insulin or insulin B chain by a variety of strategies prevents IDDM in NOD mice. Surprisingly, young NOD mice receiving i.m. injections of pDNA encoding insulin B chain-IgGFc with or without IL-4 exhibited an accelerated progression of insulitis and developed early diabetes. Exacerbation of IDDM correlated with an increased frequency of IFN-gamma-secreting CD4(+) and CD8(+) T cells in response to insulin B chain-specific peptides compared with untreated mice. In contrast, treatment with pDNAs encoding insulin A chain-IgGFc and IL-4 elicited a low frequency of IL-4-secreting Th cells and had no effect on the progression of IDDM. Vaccination with pDNAs encoding GAD65-IgGFc and IL-4, however, prevented IDDM. These results demonstrate that insulin- and GAD65 specific T cell reactivity induced by pDNA vaccination has distinct effects on the progression of IDDM. PMID- 11418699 TI - Public health medicine in transition. PMID- 11418701 TI - Medicolegal consequences of postoperative intra-abdominal adhesions. AB - Postoperative adhesions are an almost invariable consequence of abdominal and pelvic surgery. Their most important morbidity is small-bowel obstruction, but other sequelae include female infertility and dyspareunia and increased risk of visceral injury at subsequent laparotomy or laparoscopy. Whether chronic abdominal pain is truly a consequence of adhesions is debatable, although it is likely to be accepted as an entity by both patients and their legal advisors. Of 14 successful claims dealt with by a British medical defence organization, 5 were for perforations after laparoscopic division of adhesions, 2 for adhesions after laparoscopic surgery, 1 for infertility as a result of adhesions and 6 for delayed diagnosis of obstruction. General practitioners, surgeons and gynaecologists need to be aware of the increasing burden of medicolegal claims arising from these complications. PMID- 11418700 TI - Exploring the causes of adverse events in NHS hospital practice. AB - In a previous paper we reported that 10.8% of patients admitted to two large hospitals in Greater London experienced one or more adverse events, of which half were deemed preventable. Here we examine the underlying causes of errors in clinical practice. Rather than identifying specific errors made by individuals, we have looked at possible faults in the organization of care. Adverse events were grouped according to stages in the care process: diagnosis, preoperative assessment and care, operative or invasive procedure (including anaesthesia), ward management, use of drugs and intravenous fluids and discharge from hospital. Less than 20% of preventable adverse events were directly related to surgical operations or invasive procedures and less than 10% to misdiagnoses. 53% of preventable adverse events occurred in general ward care (including initial assessment and the use of drugs and intravenous fluids) and 18% in care at the time of discharge. Probable contributory factors in these errors included dependence on diagnoses made by inexperienced clinicians, poor records, poor communication between professional carers, inadequate input by consultants into day-to-day care, and lack of detailed assessment of patients before discharge. PMID- 11418702 TI - Essential healthcare for people with learning disabilities: barriers and opportunities. PMID- 11418703 TI - Histopathological aspects of cutaneous lymphoma. PMID- 11418704 TI - Management of the difficult airway in children. PMID- 11418705 TI - Demonstration of small-bowel fistula by radiography of drain contents. PMID- 11418706 TI - Thyrotoxicosis with heart block. PMID- 11418707 TI - Crutches in Cushing's disease. PMID- 11418708 TI - Difficult insulinoma. PMID- 11418709 TI - Severe adrenal suppression by steroid nasal drops. PMID- 11418710 TI - Not pre-eclampsia. PMID- 11418711 TI - An unusual pelvic mass. PMID- 11418712 TI - The flowering of London pride: finding a name for it. PMID- 11418713 TI - From convicts to founding fathers-three notable Sydney doctors. PMID- 11418714 TI - Computer game delusions. PMID- 11418715 TI - Diabetic nephropathy and dyslipidaemia. PMID- 11418716 TI - Superficial thrombophlebitis followed by pulmonary embolism. PMID- 11418717 TI - Surgical decisions in the elderly. PMID- 11418718 TI - Surgical decisions in the elderly. PMID- 11418719 TI - Why do we have apocrine and sebaceous glands? PMID- 11418720 TI - Rape in a learning-disabled woman. PMID- 11418721 TI - Leechcraft. PMID- 11418722 TI - Reducing risk by improving standards of intrapartum care. PMID- 11418723 TI - Pediatric non-Hodgkin lymphomas in children: diagnosis and current management. PMID- 11418724 TI - Isolation of measles virus from cerebrospinal fluid of children with acute encephalopathy without rash. AB - OBJECTIVE: To determine the viral agent involved in cases of acute encephalopathy in children during an outbreak in Northern India. DESIGN: Virological and serological studies using serum and cerebrospinal fluid specimens from patients. METHODS: Serum and CSF specimens were tested by IgM ELISA for IgM antibodies to variety of viruses like Japanese encephalitis, West Nile, Dengue and Measles. The specimens were inoculated into Vero cell monolayer for virus isolation. The viral strains isolated were identified by indirect immunofluorescence test and qualitative in-vitro neutralization test using polyclonal and monoclonal antibodies to measles. Identity of the isolates was reconfirmed using RT-PCR method. RESULTS: Of the 28 specimens tested, 17 had IgM antibodies to measles. Commercial IgM ELISA kits confirmed the serological findings. Vero cell cultures yielded 4 isolates from CSF and 2 from serum specimens of six different patients. Cytopathic effect was typical of measles. Indirect imunofluorescence using polyclonal and monoclonal antibodies to measles HA protein, confirmed the measles etiology. Neutralization tests reconfirmed the measles strain isolation. RT-PCR amplified product was confirmed as measles. CONCLUSION: The isolation of measles virus from CSF and serum of children with acute encephalopathy without rash proved the etiological role of measles virus in this outbreak. PMID- 11418725 TI - Early predictors of mortality in very low birth weight neonates. AB - OBJECTIVE: To evaluate early predictors of mortality in very low birth weight neonates. SETTING: Teaching hospital. DESIGN: Case control study. METHODS: Hospital born very low birth weight newborns (500-1500 g) enrolled for study and followed up till death or 28 days. Infants' birth data and data on physiologic alterations, investigation and interventions in the first 24 hours of life and CRIB score were analyzed for their ability to predict neonatal mortality. RESULTS: 115 subjects were enrolled into the study of which 47 died in the neonatal period. The factors significantly associated with early neonatal mortality included birth weight, gestation, low Apgar scores, need for assisted ventilation at birth, need for supplemental oxygen and mechanical ventilation in the first 24 hours, presence of shock, hypoxia and acidosis (p < 0.05). The factors associated with late neonatal mortality were birth weight and gestation only. Multivariate analysis of these factors showed that besides low birth weight, shock, need for mechanical ventilation, acidosis and high alveolar arterial oxygen gradients were significant predictors of neonatal mortality. When compared with the CRIB score, birth weight <1200g proved to be an equally good predictor of mortality risk. CONCLUSION: VLBW neonates with disturbed cardio pulmonary physiology during the first 24 hours of life, especially those in need of mechanical ventilation, are at an increased risk of early neonatal mortality. PMID- 11418726 TI - IAP guidelines for graduate medical education in pediatrics. PMID- 11418727 TI - Effective PubMed search. PMID- 11418728 TI - Carrier detection and prenatal diagnosis in Duchenne/Becker muscular dystrophy. PMID- 11418729 TI - Surfactant therapy for hyaline membrane disease: the Chandigarh experience. PMID- 11418730 TI - Role of intra-muscular magnesium therapy in management of persistent apnea and prevention of adverse life threatening events. PMID- 11418731 TI - Rapid manual test for falciparum malaria. PMID- 11418732 TI - Assessment of current status of salt iodization at the beneficiary level in selected districts of Uttar Pradesh, India. PMID- 11418733 TI - Langerhans cell histiocytosis involving single system. PMID- 11418734 TI - Diffuse mesangial sclerosis presenting as infantile nephrotic syndrome. PMID- 11418735 TI - Short bowel syndrome: unusual cause of vitamin [correction of vitmain] K deficiency. PMID- 11418736 TI - Trichosporonosis in a previously healthy child in Nepal. PMID- 11418737 TI - Eosinophilia, pleural effusion and cysticercosis--unknown association? PMID- 11418738 TI - Neurofibromatosis-1 in a family. PMID- 11418739 TI - Proteus syndrome. PMID- 11418740 TI - Recurrent HIV parotitis. PMID- 11418741 TI - Mother-to-child transmission of HIV must be prevented. PMID- 11418742 TI - Strategy for preventing vertical transmission of HIV. PMID- 11418744 TI - Pediatricians in Nepal. PMID- 11418745 TI - Enzyme replacement therapy in Gaucher's disease. PMID- 11418746 TI - Geometrical theory of triple junctions of CSL boundaries. AB - When three grain boundaries having misorientations generating coincidence site lattices (CSLs) meet at a triple junction, a common (triple-junction) CSL is formed. A theory is developed as a set of theorems establishing the relationships between the geometrical parameters of the grain-boundary and triple-junction CSLs. Application of the theory is demonstrated in detail for the case of the cubic crystal system. It is also shown how the theory can be extended to an arbitrary crystal lattice. PMID- 11418747 TI - Introduction to a general crystallography. AB - The definition of an extended crystallographic group is given, based on an n dimensional Euclidean space, carrier of a faithful integral representation of a permutation group of atomic positions. The Euclidean crystallography of normal crystals and the higher-dimensional one applied to incommensurately modulated crystals, intergrowth crystals and quasicrystals are special cases of a general crystallography. The same is true for the multimetrical crystallographic characterization of ice and of snow crystals. This approach can also be applied to single molecules, leading to what may be denoted as molecular crystallography. It possibly allows non-trivial structural relations between atomic positions belonging to the asymmetric unit of the molecular point group to be obtained. Two simple molecules, polycyclic aromatic hydrocarbons, are treated as illustrative examples. PMID- 11418748 TI - Approximate solution of the Takagi-Taupin equations for a semi-infinite crystal in the three-beam Laue-Laue case. AB - An analytical approximate solution of the Takagi-Taupin equations for a symmetrical three-beam Laue-Laue case in a perfect non-absorbing semi-infinite crystal slab has been obtained. The expression, a second-order expansion, is valid for situations where the effective crystal thickness does not exceed half the actual extinction length and it is shown to be in perfect agreement with the full numerical solution of the fundamental equations. PMID- 11418749 TI - Critical examination of the radial functions in the Hansen-Coppens multipole model through topological analysis of primary and refined theoretical densities. AB - A double-zeta (DZ) multipolar model has been applied to theoretical structure factors of four organic molecular crystals as a test of the ability of the multipole model to faithfully retrieve a theoretical charge density. The DZ model leads to significant improvement in the agreement with the theoretical charge density along the covalent bonds and its topological parameters, and eliminates some of the bias introduced by the limited flexibility of the radial functions when a theoretical density is projected into the conventional multipole formalism. The DZ model may be too detailed for analysis of experimental data sets of the accuracy and resolution typically achieved at present, but provides guidance for the type of algorithms to be adapted in future studies. PMID- 11418750 TI - A proposal for a transition mechanism from the diamond to the lonsdaleite type. AB - A phase transition between the diamond (Fd3;m) and the lonsdaleite types (P6(3)/mmc) may be described as a deformation of a homogeneous sphere packing with three contacts per sphere (type 3/10/o1) in the common subgroup Pnna of Fd3;m and P6(3)/mmc. The frequently observed transition between the zinc-blende (F4;3m) and the wurtzite types (P6(3)mc) may be described in an analogous way as a deformation of a heterogeneous sphere packing in the subgroup Pna2(1). The proposed model guarantees the three-dimensional connection during the whole transformation process. By this property it is distinguished from other models. PMID- 11418751 TI - The method of joint probability distribution functions applied to MAD techniques. The centric case. AB - Traditional probabilistic approaches consider MAD (multiple-wavelength anomalous dispersion) data as a special MIR (multiple isomorphous replacement) case. The rigorous method of the joint probability distribution functions has been applied to solve the phase problem, with the assumption that the anomalous scatterers' substructure is a priori known. The probabilistic approach is able to handle measurement errors: it has been applied to symmetry-restricted phases and provides simple and efficient formulas. PMID- 11418752 TI - Phase determination and extension using X-ray multiple diffraction and the maximum-entropy method. AB - The extension of the phases of the structure factors of the organic crystal C(25)H(25)NO(2) from 77 starting individual phases using the maximum-entropy method is reported. These starting phases were determined from 90 experimental triplet phases calculated from 215 measured psi-scan three-beam and four-beam diffraction profiles obtained with a rotating-anode X-ray source, where the psi scans were around the reciprocal-lattice vectors of the 001, 002 and 003 reflections. The extension of the structure factors with phase values was carried out using the maximum-entropy method for 2040 measured two-beam Bragg diffraction intensities with 77 starting phases and the symmetry of the space group as the constraints. Use of structure-factor triplets as constraints for entropy maximization was also attempted. The minimum chi(2) criteria were applied to the maximum-entropy extrapolation to discern the best phase set to be used as the new constraints for the next step of generating new phases. With this phase-extension procedure, more than 100 phases were determined and an electron-density map at 1.97 A was deduced. PMID- 11418753 TI - Rapid neutron-diffraction data collection for hydrogen-bonding studies: application of the Laue diffractometer (LADI) to the case study zinc (tris)thiourea sulfate. AB - The successful application of the newly developed image-plate neutron Laue diffractometer (LADI) at the Institut Laue-Langevin (ILL), Grenoble, France, for rapid hydrogen-bonding characterization is reported. The case study concerns the promising non-linear optical material zinc (tris)thiourea sulfate (ZTS), which contains 30 atoms in the asymmetric unit and crystallizes in the orthorhombic space group, Pca2(1), a = 11.0616 (9), b = 7.7264 (6), c = 15.558 (1) A [T = 100.0 (1) K]. The results from a 12 h data collection from ZTS on LADI are compared with those obtained over 135 h using the monochromatic four-circle diffractometer D9 at the same reactor source with a crystal 13 times larger in volume. Both studies reveal the extensive hydrogen bonding and other close non bonded contacts within the material. As expected, the results from D9 are more precise than those obtained from LADI; however, the bond geometry determined from the two experiments is the same within the larger estimated standard deviations. Furthermore, the conclusions drawn from the two studies separately regarding the nature of all supramolecular features are identical. This illustrates that LADI is eminently suitable for rapid characterization of hydrogen-bonded structures by neutron diffraction, with the gain in speed compared with traditional instrumentation being several orders of magnitude. PMID- 11418754 TI - Latent symmetry and its group theoretical determination. AB - Latent symmetry of a component of a composite object is defined as that symmetry of a subunit of the component which is not a symmetry of the component but which is a symmetry of the composite. This is illustrated by a geometrical example. A group theoretical procedure is given to determine latent symmetry and is applied to examples. PMID- 11418755 TI - Derivation of a scoring function for crystal structure prediction. AB - The ever increasing number of experimentally resolved crystal structures supports the possibility of fully empirical crystal structure prediction for small organic molecules. Empirical methods promise to be significantly more efficient than methods that attempt to solve the same problem from first principles. However, the transformation from data to empirical knowledge and further to functional algorithms is not trivial and the usefulness of the result depends strongly on the quantity and the quality of the data. In this work, a simple scoring function is parameterized to discriminate between the correct structure and a set of decoys for a large number of different molecular systems. The method is fully automatic and has the advantage that the complete scoring function is parametrized at once, leading to a self-consistent set of parameters. The obtained scoring function is tested on an independent set of crystal structures taken from the P1 and P1; space groups. With the trained scoring function and FlexCryst, a program for small-molecule crystal structure prediction, it is shown that approximately 73% of the 239 tested molecules in space group P1 are predicted correctly. For the more complex space group P1;, the success rate is 26%. Comparison with force-field potentials indicates the physical content of the obtained scoring function, a result of direct importance for protein threading where such database-based potentials are being applied. PMID- 11418756 TI - Fast differentiation algorithm and efficient calculation of the exact matrix of second derivatives. AB - Many crystallographic problems are reduced to the optimization of some functional. In most cases, this functional is expressed in terms of structure factors and depends on a large number of variables; an example is the refinement of atomic models. Calculation of the functional derivatives, necessary for different optimization methods, is a time-consuming procedure. Previously, a technique to calculate the exact gradient of any crystallographic functional for the time equal to that for a single-function-value calculation has been proposed [Lunin & Urzhumtsev (1985). Acta Cryst. A41, 327-333]. Currently, a similar scheme is proposed to calculate the exact matrix of the second derivatives of these functionals. The accuracy of this matrix is crucial for the calculation of the inverted matrix, which can be used in optimization methods of the second order. PMID- 11418757 TI - The joint probability distributions of structure-factor doublets in displacive incommensurately modulated structures and their applicability to direct methods. AB - In 1993, alternative normalized structure factors for incommensurately modulated structures were defined [Lam, Beurskens & van Smaalen (1993). Acta Cryst. A49, 709-721]. The probability distribution associated with the structure invariants E(-H)E(H')E(H - H') has approximately the same functional form as the Cochran distribution. It was shown, however, that triplet-phase relations are relatively less reliable when satellites are involved [de Gelder, Israel, Lam, Beurskens, van Smaalen, Fu & Fan (1996). Acta Cryst. A52, 947-954]. In the present paper, an alternative approach is presented: instead of studying the distribution of a three-phase invariant, the probability distribution of the phase sum of two first order satellite reflections (h,k,l,1 and h',k',l',-1) has been derived under the assumption that the phase of the associated main reflection (h + h',k + k',l + l',0) can be calculated from the known main (or averaged) structure. Intensive tests with randomly generated artificial structures and one real structure show a significant improvement of direct-methods phase-sum statistics. Functional similarities with conventional direct methods, employing normalized structure factors and the Cochran distribution, are discussed. PMID- 11418758 TI - Inversion of dynamical electron scattering to obtain the crystal potential using data from two thicknesses. AB - A method is given to invert the multiple scattering of electrons in a crystalline slab to obtain the projected potential. For a fixed orientation of the incident beam of electrons, scattering data are required for two thicknesses of the crystal. PMID- 11418759 TI - The Fedorov algorithm revised. AB - All the simple 12- to 20-hedra with no triangular and quadrilateral facets (118 in common) are calculated in the Schlegel projections. Among them, all the fullerenes with 20 to 36 vertices (35 in common) are found. Thus, the gamma operation is eliminated from the Fedorov algorithm up to a 21-hedra generating procedure. PMID- 11418760 TI - Structural effects of monovalent anions on polymorphic lysozyme crystals. AB - Understanding direct salt effects on protein crystal polymorphism is addressed by comparing different crystal forms (triclinic, monoclinic, tetragonal and orthorhombic) for hen, turkey, bob white quail and human lysozymes. Four new structures of hen egg-white lysozyme are reported: crystals grown in the presence of NapTS diffracted to 1.85 A, of NaI to 1.6 A, of NaNO(3) to 1.45 A and of KSCN to 1.63 A. These new structures are compared with previously published structures in order to draw a mapping of the surface of different lysozymes interacting with monovalent anions, such as nitrate, chloride, iodide, bromide and thiocyanate. An analysis of the structural sites of these anions in the various lysozyme structures is presented. This study shows common anion sites whatever the crystal form and the chemical nature of anions, while others seem specific to a given geometry and a particular charge environment induced by the crystal packing. PMID- 11418761 TI - Intermonomer interactions in dimer of bovine heart cytochrome c oxidase. AB - The X-ray structure of bovine heart cytochrome c oxidase solved for orthorhombic crystals showed a dimeric structure stabilized by four subunit-subunit contacts, namely, subunit Vb-subunit Vb on the matrix side, subunit I-subunit VIa, subunit VIa-subunit I in the transmembrane region and subunit VIb-subunit VIb on the intermembrane side. The same intermonomer contacts as in the orthorhombic crystals were observed in both hexagonal and tetragonal crystals, the X-ray structures of which were determined by the molecular-replacement method. These results suggest that the dimeric structure also exists under physiological conditions. These contacts, especially the subunit IVa-subunit I contact, in which the N-terminal portion of subunit IVa is placed on the surface of subunit I near the dioxygen-reduction site, indicate that the function of the bovine heart enzyme is likely to be controlled by perturbation of the monomer-monomer association. PMID- 11418762 TI - Structure of aspergillopepsin I from Aspergillus phoenicis: variations of the S1' S2 subsite in aspartic proteinases. AB - The crystal structure of aspergillopepsin I (AP) from Aspergillus phoenicis has been determined at 2.18 A resolution and refined to R and R(free) factors of 21.5 and 26.0%, respectively. AP has the typical two beta-barrel domain structure of aspartic proteinases. The structures of the two independent molecules are partly different, exemplifying the flexible nature of the aspartic proteinase structure. Notably, the 'flap' in one molecule is closer, with a largest separation of 4.0 A, to the active site than in the other molecule. AP is most structurally homologous to penicillopepsin (PP) and then to endothiapepsin (EP), which share sequence identities of 68 and 56%, respectively. However, AP is similar to EP but differs from PP in the combined S1'-S2 subsite that is delineated by a flexible psi-loop in the C-terminal domain. The S1' and S2 subsites are well defined and small in AP, while there is no definite border between S1' and S2 and the open space for the S2 subsite is larger in PP. Comparison of the structures indicates that the two amino-acid residues equivalent to Leu295 and Leu297 of AP are the major determining factors in shaping the S1'-S2 subsite in the fungal aspartic proteinases. PMID- 11418763 TI - Structure of a new polymorphic monoclinic form of human transthyretin at 3 A resolution reveals a mixed complex between unliganded and T4-bound tetramers of TTR. AB - The crystal structure of a new polymorphic form of human transthyretin (hTTR) with a lattice containing a unique assembly of apo hTTR and TTR-T(4) complex has been determined to 3 A resolution. The monoclinic form of human TTR reported here crystallizes in space group P2(1), with unit-cell parameters a = 76.7 (6), b = 96.7 (8), c = 81.7 (4) A, beta = 106.8 (4) degrees. The asymmetric unit contains two tetramers of transthyretin related by the non-crystallographic symmetry (NCS) operation of a 90.28 degrees rotation between two hTTR molecules around an axis close to crystallographic z. The r.m.s. difference between the two tetramers calculated from their C(alpha) positions is 0.48 A. The structure was refined using 15.0-3.0 A resolution data to R = 22.9% and R(free) = 28.9% for reflections F > 0.0sigma(F), and R = 19.7% and R(free) = 25.8% for reflections F > 3.0sigma(F). The intermolecular interactions involve the tips of alpha-helices and loops around Arg21, Glu61 and Ser100 of all monomers. The electron-density maps revealed residual thyroxine (T(4)) bound in only one of the two unique tetrameric TTR molecules, with an occupancy of 53%, while the second tetramer is unliganded. One thyroxine ligand is bound in a way similar to the orientations described for the orthorhombic form of the hTTR-T(4) complex. The T(4) bound in the second site is positioned similar to 3',5'-dinitro-N-acetyl-L-thyronine in its hTTR complex. Differences in the size of the central channel defined by the D, A, G and H beta-strands of two monomeric subunits are observed between the apo TTR and T(4)-bound tetramer. The averaged distances between Ala108 C(alpha) and its equivalent measured across each binding site are 12.34 A for the T(4)-bound and 10.96 A for the unliganded TTR tetramer, respectively. The observed differences might reflect the mechanics of the ligand binding in the channel and possibly explain the observed negative cooperativity effect for ligand binding. PMID- 11418764 TI - Structure of ribosomal protein TL5 complexed with RNA provides new insights into the CTC family of stress proteins. AB - The crystal structure of Thermus thermophilus ribosomal protein TL5 in complex with a fragment of Escherichia coli 5S rRNA has been determined at 2.3 A resolution. The protein consists of two domains. The structure of the N-terminal domain is close to the structure of E. coli ribosomal protein L25, but the C terminal domain represents a new fold composed of seven beta-strands connected by long loops. TL5 binds to the RNA through its N-terminal domain, whereas the C terminal domain is not included in this interaction. Cd(2+) ions, the presence of which improved the crystal quality significantly, bind only to the protein component of the complex and stabilize the protein molecule itself and the interactions between the two molecules in the asymmetric unit of the crystal. The TL5 sequence reveals homology to the so-called general stress protein CTC. The hydrophobic cores which stabilize both TL5 domains are highly conserved in CTC proteins. Thus, all CTC proteins may fold with a topology close to that of TL5. PMID- 11418765 TI - De novo structure determination of vancomycin aglycon using the anomalous scattering of chlorine. AB - The crystal structure of vancomycin aglycon has been determined by exploiting the anomalous scattering of Cl atoms present within the molecule. Real-space reciprocal-space cycling with Shake-and-Bake successfully located the chlorine positions from the Bijvoet differences, even though the anomalous difference Patterson map proved to be uninterpretable. The chlorine anomalous differences lacked sufficient phasing power to produce interpretable electron-density maps. However, when combined with high-resolution native data, the chlorine positions were sufficient to determine the structure using either Shake-and-Bake or a tangent-formula expansion. PMID- 11418766 TI - High-resolution neutron structure of nicotinamide adenine dinucleotide. AB - The structure of the free-acid form of the coenzyme NAD(+) was determined at 100 K from a single-crystal neutron experiment. NAD(+) is the oxidized form of the coenzyme redox pair NAD(+)/NADH and plays an important role in the catalysis of biological processes. The molecule crystallizes in space group P1 with one NAD(+) and four water molecules per unit cell. The structure is compared with the previous X-ray models of NAD(+) [Reddy et al. (1981), J. Am. Chem. Soc. 103, 907 914; Parthasarathy & Fridey (1984b), Science, 226, 969-971; Guillot et al. (2000), Acta Cryst. C56, 726-728]. The crystal packing and the hydrogen-bond pattern are discussed as well as four short C-H.O contacts involving the pyridine and adenine rings. The structure displays stereochemical distortions owing to the hydrogen bonding and crystal-packing constraints, reflecting the adaptability of the NAD(+) molecule in various chemical environments. PMID- 11418767 TI - Anomalous signal of phosphorus used for phasing DNA oligomer: importance of data redundancy. AB - The crystal structure of a left-handed Z-DNA hexamer duplex d(CGCGCG)(2) has been solved based on the anomalous diffraction signal of inherent P atoms using data collected at the single wavelength of 1.54 A. The anomalous signal of about 2% of the total diffracted intensity, constant for all nucleic acids, may be generally useful for solving crystal structures of DNA and RNA oligomers. The multiplicity of intensity measurements is shown to crucially affect the data quality and the ability to solve the phase problem. The anisotropic model refined to an R factor of 8.9% at 0.95 A resolution. PMID- 11418768 TI - Protein crystal structure solution by fast incorporation of negatively and positively charged anomalous scatterers. AB - The preparation of derivatives by the traditional methods of soaking is one of the most time-consuming steps in protein crystal structure solution by X-ray diffraction techniques. The 'quick cryosoaking' procedure for derivatization with halides (monovalent anions) offers the possibility of significantly speeding up this process [Dauter et al. (2000), Acta Cryst. D56, 232-237]. In the present work, an extension of this technique is proposed and the use of two different classes of compounds (monovalent and polyvalent cations) that can be successfully utilized in the quick cryosoaking procedure for the derivatization and phasing of protein crystals is described. This approach has been tested on hen egg-white lysozyme and has been successfully used to solve the structure of a novel trypsin inhibitor. The possibility of using cations in the fast cryosoaking procedure gives additional flexibility in the process of derivatization and increases the chances of success in phase determination. This method can be applied to high throughput crystallographic projects. PMID- 11418769 TI - The absorption edge of protein-bound mercury and a double-edge strategy for HgMAD data acquisition. AB - The L(III) absorption edge of protein-bound mercury (Hg) has been experimentally determined using X-ray data collection from a crystal. This absorption edge is 12 291 eV, 4 eV higher than the theoretical value of elemental Hg. Considering the possible shift of the Hg absorption edge with the chemical environment in different protein crystals, a double-edge strategy for multiwavelength anomalous diffraction (MAD) data collection has been developed. The approach provides a convenient way to optimize the dispersive signal between a remote wavelength and two edge wavelengths separated from each other by 4 eV. The dispersive signals derived from both edges are used, along with anomalous signals, in MAD phasing and phase refinement. This approach has been used in the crystal structure determination of three proteins containing one Hg atom per 186-196 amino-acid residues at 2.0, 2.6 and 2.7 A resolution. A set of four wavelengths is recommended for HgMAD data acquisition: 1.0087 A (12 291 eV, edge1), 1.0084 A (12 295 eV, edge2), 1.0064 A (12 320 eV, peak) and 0.9918 A (12 500 eV, remote). Although it is no longer necessary to determine the L(III) absorption edge of protein-bound Hg experimentally, an initial fluorescence scan on the crystal for data collection is still necessary to verify the existence of Hg in the crystal. PMID- 11418770 TI - Using surface-bound rubidium ions for protein phasing. AB - Rubidium is a monovalent metal that can be used as a counterion in protein solutions. X-ray anomalous scattering from rubidium ions bound to the protein surface was used for phasing of the crystal structure of the hsp60 apical domain from Thermus thermophilus. Multiple-wavelength anomalous dispersion (MAD) data were collected from a crystal obtained from a solution containing 0.2 M rubidium salt. One molecule of protein (147 amino acids) binds one well ordered and one poorly ordered Rb atom. Phases calculated with the program SHARP were sufficient for automatic tracing and side-chain assignment using the program ARP/wARP. The data show that bound rubidium ions can be used to determine protein structures and to study the interaction of monovalent metal ions with proteins and other macromolecules. PMID- 11418771 TI - A new software routine that automates the fitting of protein X-ray crystallographic electron-density maps. AB - The classical approach to building the amino-acid residues into the initial electron-density map requires days to weeks of a skilled investigator's time. Automating this procedure should not only save time, but has the potential to provide a more accurate starting model for input to refinement programs. The new software routine MAID builds the protein structure into the electron-density map in a series of sequential steps. The first step is the fitting of the secondary alpha-helix and beta-sheet structures. These 'fits' are then used to determine the local amino-acid sequence assignment. These assigned fits are then extended through the loop regions and fused with the neighboring sheet or helix. The program was tested on the unaveraged 2.5 A selenomethionine multiple-wavelength anomalous dispersion (SMAD) electron-density map that was originally used to solve the structure of the 291-residue protein human heart short-chain L-3 hydroxyacyl-CoA dehydrogenase (SHAD). Inputting just the map density and the amino-acid sequence, MAID fitted 80% of the residues with an r.m.s.d. error of 0.43 A for the main-chain atoms and 1.0 A for all atoms without any user intervention. When tested on a higher quality 1.9 A SMAD map, MAID correctly fitted 100% (418) of the residues. A major advantage of the MAID fitting procedure is that it maintains ideal bond lengths and angles and constrains phi/psi angles to the appropriate Ramachandran regions. Recycling the output of this new routine through a partial structure-refinement program may have the potential to completely automate the fitting of electron-density maps. PMID- 11418772 TI - Effects of PEG on detergent micelles: implications for the crystallization of integral membrane proteins. AB - The solubilization of integral membrane proteins with detergents produces protein detergent complexes (PDCs). Interactions between the detergent moieties of PDCs contribute significantly to their behavior. The effects of the precipitating agent polyethylene glycol (PEG) upon these detergent-detergent interactions have been examined, focusing on the detergent system used to crystallize the bacterial outer membrane protein OmpF porin. Static and dynamic light scattering were used to assess the effects of temperature and concentration upon the hydrodynamic size distribution and the aggregation state of detergent micelles and a phase diagram for micellar solutions was mapped. Estimates of the second osmotic virial coefficient obtained from static light-scattering measurements on micelles were shown to accurately reflect the thermodynamic quality of the solvent. Solvent quality decreases as the consolute boundary is approached, suggesting micelle micelle attractive forces help to organize PDCs into crystalline aggregates near the cloud point. An apparent increase in micelle mass is observed as the solution approaches the cloud point. These results raise the possibility that the detergent-mediated aggregation of PDCs and/or slight changes in micelle geometry may prove to be important in the nucleation of membrane protein crystals. PMID- 11418773 TI - Preliminary crystallographic study of Thermus aquaticus glycerol kinase. AB - Glycerol kinase (GlpK) is an important enzyme which catalyzes the rate-limiting step in a central biochemical pathway involving glycerol metabolism. GlpK from the thermophile Thermus aquaticus has been overexpressed in glpK-deficient Escherichia coli and crystallized by the hanging-drop method. The crystal belongs to the cubic space group I23, with unit-cell parameters a = b = c = 163.94 (3) A. Native data were collected to 2.87 A resolution on a Cu Kalpha rotating-anode X ray source. PMID- 11418774 TI - Crystallization and preliminary X-ray analysis of UDP-N acetylenolpyruvylglucosamine reductase (MurB) from Staphylococcus aureus. AB - UDP-N-acetylenolpyruvylglucosamine reductase (MurB) is an essential enzyme in the bacterial cell-wall biosynthetic pathway, making it a potential therapeutic target for novel antibiotics. Diffraction-quality crystals of both the native and Se-methionine-expressed MurB from Staphylococcus aureus have been prepared by sitting-drop vapour diffusion from solutions containing polyethylene glycol (PEG) 8000, ammonium sulfate, sodium cacodylate pH 6.5 and dimethyl sulfoxide (DMSO). Crystals belong to the cubic space group I2(1)3, with unit-cell parameters a = b = c = 178.99 A. X-ray data from these crystals were collected at the Advanced Photon Source 17-ID beamline and were used to solve the MurB structure to 2.3 A resolution. PMID- 11418775 TI - Crystallization and X-ray diffraction measurements of a thermophilic archaeal recombinant amidase from Sulfolobus solfataricus MT4. AB - Recombinant amidase is a 55.8 kDa enzyme from the thermophilic archaeon Sulfolobus solfataricus MT4 that catalyses the hydrolysis of aliphatic amides of 2-6 C atoms as well as many aromatic amides. Single crystals of purified amidase were obtained by the hanging-drop method at 294 K. Diffraction data for the native protein (2.55 A resolution) and a putative derivative (2.20 A) have been collected at low temperature using synchrotron radiation. The crystals belong to the rhombohedral space group R3. Structure determination by multiple isomorphous replacement is in progress. It is expected that structural information from this signatured thermostable amidase will increase our knowledge of the molecular mechanisms employed to maintain high-temperature stability in thermophilic proteins. PMID- 11418776 TI - Cloning, purification, crystallization and preliminary X-ray diffraction analysis of the antistasin-type inhibitor ghilanten (domain I) from Haementeria ghilianii in complex with porcine beta-trypsin. AB - Ghilanten, isolated from the leech Haementeria ghilianii, is a potent two-domain anticoagulant protein homologous to the factor Xa inhibitor antistasin. A synthetic gene encoding the amino-terminal domain of ghilanten (ghilanten-D1) was constructed, expressed in the methylotrophic yeast Pichia pastoris and purified by heparin-Sepharose chromatography. Recombinant ghilanten-D1 inhibits bovine trypsin and human factor Xa with equilibrium inhibition constants (K(i)) of 126 and 1.2 nM, respectively. Ghilanten-D1 has been crystallized in complex with porcine beta-trypsin; three different-looking but isomorphous crystal forms were obtained, each belonging to the orthorhombic space group P2(1)2(1)2(1). These crystals diffracted to beyond 3.6 A resolution using a rotating-anode X-ray source. A data set complete to 3.7 A resolution was collected. PMID- 11418777 TI - Characterization and crystallization of a novel Sarcocystis muris lectin, SML-2. AB - A novel lectin (SML-2) consisting of 138 amino acids was isolated from cyst merozoites of Sarcocystis muris and sequenced by Edman degradation and mass spectrometry. All 12 cysteinyl residues are involved in disulfide bridges, four of which are attributed to a characteristic pattern of cysteines as found in the so-called PAN-module superfamily. Crystals of SML-2 diffracting to 2.1 A resolution at a synchrotron were grown by the hanging-drop vapour-diffusion technique. They belong to the space group P2(1)2(1)2(1), with unit-cell parameters a = 53.6, b = 128.8, c = 158.2 A and eight molecules in the asymmetric unit. SML-2 cocrystallized with Au galactose results in two different crystal forms. The first form is isomorphous with the native crystals and the second form adopts space group C222(1), with unit-cell parameters a = 74.7, b = 82.0, c = 131.0 A, and diffracts to 2.4 A at a rotating-anode X-ray generator. PMID- 11418778 TI - Crystallization and preliminary X-ray crystallographic studies on 4-alpha glucanotransferase from Thermotoga maritima. AB - Thermotoga maritima 4-alpha-glucanotransferase (GTase), a 52 kDa molecular-weight amylolytic enzyme, has been crystallized by the hanging-drop vapour-diffusion method using PEG monomethylether 5000 as a precipitating agent. A complete data set has been collected to 2.6 A resolution using cryocooling conditions and synchrotron radiation. The crystals belong to space group I222 or I2(1)2(1)2(1), with unit-cell parameters a = 92.6, b = 180.3, c = 199.2 A. PMID- 11418779 TI - An improved phase-extension procedure for isomorphous replacement phases. AB - A new phase-extension procedure has been applied to isomorphous replacement data and shown to yield improved phases and maps compared with standard solvent flattening operating on a full set of centroid phases. In this procedure, a starting subset of core phases is selected based on the sharpness of the phase probability curves. Phase extension using solvent flattening as the density modification procedure is then carried out, gradually adding additional phases. In tests with known protein structures, the mean phase errors for the output expanded phase sets were reduced by 3-9 degrees and the corresponding map correlation coefficients were increased by 0.05-0.18 relative to phase sets from standard solvent-flattening procedures. With SIR data, the lowest final mean phase errors were approximately 58 degrees and the corresponding map correlation coefficients were in the range 0.53-0.68. PMID- 11418781 TI - Increasing concerns about chronic proton pump inhibitor use. PMID- 11418782 TI - Diffuse esophageal spasm: a case for carminatives? PMID- 11418783 TI - Techniques for the evaluation of dyspepsia in children. AB - Dyspepsia can describe a subset of children with episodic or persistent abdominal symptoms--often related to feeding--that are thought to be caused by disorders of the proximal part of the digestive tract. Symptoms, such as vomiting, early satiety, postprandial epigastric abdominal pain, heartburn, abdominal fullness, poor weight gain, and/or anorexia, have been incorporated into the definition of dyspepsia. Unfortunately, presenting signs and symptoms in children with dyspepsia are nonspecific and can occur as a result of many diseases, such as parasitic infections, esophagitis, eosinophilic gastroenteritis, Helicobacter pylori infection, Crohn's disease, biliary tract or hepatic disease, pancreatitis, and lactose intolerance. This lack of specificity makes the evaluation of dyspepsia more difficult. Here, we describe an approach for the evaluation of dyspepsia that correlates in part with the child's presenting symptoms. PMID- 11418784 TI - Should indirect calorimetry be used as part of nutritional assessment? AB - The use of indirect calorimetry in the design of nutritional support regimens is poorly appreciated by clinicians, who fail to recognize the importance of providing a sufficient volume of enteral feeding to critically ill patients. In contrast to the overfeeding that routinely occurred in the past with the provision of total parenteral nutrition, patients placed on the enteral route of support tend to be underfed because of problems with intolerance and frequent cessation. Clearly identifying and coming as close as possible to the caloric goal may be required to achieve the therapeutic endpoints of enteral tube feeding (which include maintenance of gut integrity, attenuation of the stress response, prophylaxis against stress-induced gastropathy, and stimulation of immune function). Indirect calorimetry is a convenient, accessible, and highly accurate instrument for the measurement of caloric requirements and is a valuable tool for the optimization of nutritional support in the intensive care unit. PMID- 11418785 TI - Risks associated with hepatitis A and hepatitis B in patients with hepatitis C. AB - Individuals with hepatitis C virus (HCV) are at risk for acquiring hepatitis A virus (HAV) or hepatitis B virus (HBV) because of shared risk factors. A number of organizations recommend vaccination against HAV and HBV for patients with HCV. The rationale for vaccinating these patients is to prevent hepatic superinfections. Acute HAV superinfection causes more severe disease, acute hepatic failure, and higher fatality rates in patients with underlying chronic liver disease, specifically chronic HBV infection and chronic HCV infection. Available data, although limited, suggest that HBV coinfection with HAV and HCV causes more severe hepatic injury than infection with HAV or HCV alone. At standard doses, hepatitis A and hepatitis B vaccines are safe and immunogenic in patients with mild-to-moderate hepatitis C or chronic liver disease. Regardless of disease severity, vaccination should be routinely administered to patients upon diagnosis of HCV infection. Early vaccination is important because response to vaccination is reduced as liver disease progresses. Prevaccination and postvaccination serology testing is recommended in specific populations. A new combination hepatitis A and hepatitis B vaccine has been shown to be as safe and effective as monovalent hepatitis A and B vaccines and is currently under review by the United States Food and Drug Administration. A combination vaccine would offer ease of administration and convenience and could increase compliance in patients with hepatitis C or other chronic liver disease: two groups that should be more aggressively targeted by healthcare professionals. PMID- 11418786 TI - Peppermint oil improves the manometric findings in diffuse esophageal spasm. AB - BACKGROUND: Diffuse esophageal spasm (DES) is an uncommon condition that results in simultaneous esophageal contractions. Current medical treatment of DES is frequently unsatisfactory. We hypothesized that, as a smooth muscle relaxant, peppermint oil may improve the manometric findings in DES. STUDY: Eight consecutive patients with chest pain or dysphagia and who were found to have DES were enrolled during their diagnostic esophageal manometry. An eight-channel perfusion manometry system was used. Lower esophageal sphincter pressure and contractions of the esophageal body after 10 wet swallows were assessed before and 10 minutes after the ingestion of a solution containing five drops of peppermint oil in 10 mL of water. Each swallow was assessed for duration (seconds), amplitude (mm Hg), and proportion of simultaneous and multiphasic esophageal contractions. RESULTS: Lower esophageal sphincter pressures and contractile pressures and durations in both the upper and lower esophagus were no different before and after the peppermint oil. Peppermint oil completely eliminated simultaneous esophageal contractions in all patients (p < 0.01). The number of multiphasic, spontaneous, and missed contractions also improved. Because normal esophageal contractions are characteristically uniform in appearance, variability of esophageal contractions was compared before and after treatment. The variability of amplitude improved from 33.4 +/- 36.7 to 24.9 +/- 11.0 mm Hg (p < 0.05) after the peppermint oil. The variability for duration improved from 2.02 +/- 1.80 to 1.36 +/- 0.72 seconds (p < 0.01). Two of the eight patients had chest pain that resolved after the peppermint oil. CONCLUSIONS: This data demonstrates that peppermint oil improves the manometric features of DES. PMID- 11418787 TI - Effect of long-term, continuous versus alternate-day omeprazole therapy on serum gastrin in patients treated for reflux esophagitis. AB - BACKGROUND: Proton pump inhibitors have been proven to have a major role in the management of peptic diseases, especially the long-term control of reflux esophagitis. The potent inhibitory effect of omeprazole on gastric acid secretion is frequently associated with hypergastrinemia, and gastrin and its intermediates have been reported to promote gastrointestinal cellular functions and cell growth. Experimental data suggest that gastrin may affect the proliferation of colon cells and some other cancer cells. However, so far the direct role of gastrin in tumorigenesis is unclear. Although most clinical studies on long-term treatment with omeprazole or other proton pump inhibitors do not report serious adverse effects, the issue of prolonged hypergastrinemia and tissue growth is unsettled, and many clinicians are reluctant to recommend long-term use of omeprazole or of other proton pump inhibitors. STUDY: We examined the effect of long-term omeprazole treatment on serum gastrin levels in patients with reflux esophagitis when given either 20 mg daily (group 1) or on alternate days (group 2). During the follow-up period, clinical remission was monitored and maintained in all patients in group 1 and in the majority of patients in group 2. RESULTS: The mean serum gastrin level was significantly elevated in group 1 (mean +/- SE, 159 +/- 23.6 pg/mL; range, 45-620 pg/mL; n = 31) as compared with the alternate day treatment group (group 2) (66 +/- 4.8 pg/mL; range, 37-115 pg/mL; n = 21) (p < 0.005). In controls, serum gastrin levels showed similar values to those found in group 2 (54 +/- 4.3 pg/mL; range, 27-94 pg/mL; n = 20). Fourteen patients (45%) in group 1 had serum gastric ranging from 140 to 620 pg/mL, and 8 (25%) had a 6-fold or greater increase in serum gastrin. The follow-up treatment period ranged between 3 and 60 months (mean +/- SE, 16.1 +/- 2.1 months) for group 1 and 3-36 months (9.7 +/- 1.4 months) for group 2. Upon multivariate adjustment for age and duration of treatment, a significantly lower mean serum gastrin level was observed in the alternate-day group as compared with the daily treated group. CONCLUSION: Alternate-day, long-term treatment with omeprazole may be adequate to maintain remission in patients with reflux esophagitis. This regimen can assure serum gastrin levels within the normal range, thus reducing the potential risk of prolonged, sustained hypergastrinemia and profound hypochlorhydria. PMID- 11418788 TI - Improving the serum D-xylose test for the identification of patients with small intestinal malabsorption. AB - BACKGROUND: D-Xylose absorption testing is a simple, low-cost method of screening for small intestinal malabsorption. The optimum method to measure D-xylose absorption (serum vs. urine testing) is uncertain. GOALS: We present a method of improving the accuracy of D-xylose testing. STUDY: Fifty-one consecutive patients (40 with chronic diarrhea and 5 asymptomatic patients with renal insufficiency) and 6 volunteers with normal renal function were recruited. All received D xylose, 10 g intravenously and 25 mg orally, on two separate occasions. Serum concentration was determined at baseline and at frequent times thereafter. Area under the curve was calculated to infinity, and D-xylose bioavailability (F) was calculated. A nonlinear model used to derive the relationship between 3-hour D xylose concentrations and F showed that a value of less than 22.5 mg/dL correlated with an F of less than 60% (malabsorption of D-xylose). A 1-hour D xylose of less than 20 mg/dL was considered abnormal. RESULTS: Using these indexes for normal 1-and 3-hour D-xylose levels, 90% of patients with D-xylose malabsorption were identified (sensitivity, 90%; specificity, 95%), which represents a marked improvement within the conventional 1-hour D-xylose of less than 20 mg/dL alone (sensitivity, 71%; specificity, 100%). The model was applied prospectively to 15 additional patients with chronic diarrhea. Of these, 12 patients with an F of less than 60% were identified, including 2 patients with normal 1-hour D-xylose levels. CONCLUSIONS: Thus, the addition of a 3-hour D xylose serum level of less than 22.5 mg/dL to conventional 1-hour D-xylose determination greatly improves the D-xylose test for malabsorption screening. PMID- 11418789 TI - Gallstone pancreatitis: a community teaching hospital experience. AB - GOALS: The current study reviews the Bridgeport Hospital experience with gallstone pancreatitis (GP) and examines its incidence by race and gender. STUDY: Consecutive patients admitted with acute pancreatitis between October 1994 and October 1996 were identified using discharge diagnosis codes. Demographics and clinical information were abstracted. Patients were categorized as having definite GP, probable GP, and non-GP using the available information. RESULTS: One hundred twenty-three patients met criteria for acute pancreatitis. Of these, 40 met the criteria for definite GP; 14, probable; and 69, non-GP. The estimated incidence for acute pancreatitis was 45 per 100,000 person-years (95% CI = 41-58 per 100,000 person-years) for definite GP and was 20 per 100,000 person-years (95% CI = 14-25 per 100,000 person-years) for probably GP. Patients with definite or probable GP were predominantly white or Hispanic women and tended to be older. Only 16% of pancreatitis in black patients was associated with gallstones. Elevated alanine aminotransferase (ALT > 120 U/L) was highly specific (97%) in predicting GP, with a positive likelihood ratio of 18.3. Abdominal ultrasound was the most widely used imaging study. Complication rates were low. There was only one intrahospital death. CONCLUSIONS: The incidence of acute pancreatitis requiring hospitalization at our community hospital in 1994-1996 was 45 (95% CI = 41-58) per 100,000 person-years. Forty percent of these cases were associated with gallstones. Gallstone pancreatitis was more common among the elderly women and the white (white, non-Hispanic) population. Elevated ALT was highly specific in the prediction of GP. PMID- 11418790 TI - Hepatitis C virus RNA assays: a comparison of SuperQuant and Monitor. AB - Hepatitis C RNA testing has been used extensively to assess the efficacy of antiviral therapy and has increasingly become an integral part of clinical management of patients with chronic hepatitis C. A variety of commercially available hepatitis C virus (HCV) RNA tests are used to detect HCV RNA qualitatively or quantitatively. These commercial tests have fundamental differences that are reflected on the values they generate. We compared two widely used assays, HCV SuperQuant (SQ) and Amplicor HCV Monitor (M1 and M2), in sera of patients with chronic hepatitis C. A total of 506 sera from 79 patients were tested with both assays. The data were logarithmically transformed and analyzed by linear regression and measurement of agreement. Two hundred thirty eight sera had HCV RNA values within the dynamic range of both assays. The correlation between the assays was fair, with a correlation coefficient (r) of 0.699. Overall, SQ generated higher values than M1 with a mean difference of 0.558 log (SD = 0.624). One hundred ninety-four (38%) and 121 (24%) of the sera were below the dynamic range of M1 and SQ, respectively. Seventy-three sera, undetectable by M1, were positive by SQ. The Amplicor HCV Monitor 2.0 (M2) was performed in 66 sera. All were positive by SQ and M2, but only 38 were within the dynamic range of M1. The correlations between these tests were good (r = 0.68 0.78), but the agreement was rather poor. In conclusion, this study confirms that both SQ and M2 are more sensitive than M1. Additionally, our results show rather poor agreements between these assays. The recent attempts in standardizing the reporting of these assays should make their results more easily interchangeable. PMID- 11418791 TI - Balloon catheter-assisted endoscopic sclerotherapy for gastric fundal varices using alpha-cyanoacrylate monomer. AB - We present two patients with bleeding episodes from gastric fundal varices. The gastric fundal varices were treated by balloon catheter-assisted endoscopic sclerotherapy using alpha-cyanoacrylate monomer. The varices were successfully obliterated with no complications or hemodynamic changes to the gastrorenal shunts. This procedure may be considered a novel, feasible approach to the treatment of gastric fundal varices in the future. PMID- 11418792 TI - Intramural duodenal hematoma presenting as a complication of peptic ulcer disease. AB - We report the first case in the English literature of an intramural duodenal hematoma presenting as a complication of Helicobacter pylori-induced peptic ulcer disease. Intramural duodenal hematomas have been previously described in patients usually in the setting of blunt trauma, postendoscopic biopsy, gastrostomy placement, and hemostatic therapy and in patients with a coagulopathy or bleeding diathesis-but not as a presentation of peptic ulcer disease. It is important to recognize this complication, as surgical management may benefit patients with a duodenal hematoma. PMID- 11418793 TI - Liver penetration by a duodenal ulcer in a young woman. AB - Liver penetration is a rare but serious complication of peptic ulcer disease. We report a case of a 33-year-old woman who took large doses of nonsteroidal antiinflammatory drugs and developed a giant duodenal ulcer that penetrated into her liver. The diagnosis was based on histologic examination of endoscopic biopsies. She was initially treated with a proton pump inhibitor, but, within 5 weeks, she developed a symptomatic postbulbar stricture that required surgical correction. We also review 11 other reported cases of endoscopically and histologically diagnosed peptic ulcer penetration into the liver. PMID- 11418794 TI - Pancreaticobiliary arteriovenous malformation with common bile duct dilation in a patient with hemobilia. AB - Arteriovenous malformation of the pancreas is a very rare entity. We describe a case involving the head of the pancreas associated with progressive hemobilia bleeding from the lower part of the common bile duct. The patient was a 50-year old man with acute epigastralgia. Endoscopic retrograde cholangiopancreatography revealed hemobilia and cystic dilation of the common bile duct. Angiography demonstrated increased blood volume in the head of the pancreas and early filling of the superior mesenteric and portal veins. Abdominal pain and progressive anemia caused by hemobilia required surgical treatment. Histologic examination of the resected specimen revealed marked proliferation of the blood vessels in the pericholedochal area and the exact point of bleeding from the pancreaticobiliary arteriovenous malformation. PMID- 11418795 TI - Amebic liver abscess and human immunodeficiency virus infection: a report of three cases. AB - Invasive amebiasis rarely occurs in homosexual men and human immunodeficiency virus (HIV)-infected individuals and has not been regarded as a beacon for concomitant HIV infection. We encountered a bisexual man with a protracted course of amebic liver abscess and amebic colitis. In the presence of fever, generalized lymphadenopathy, and elevated serum aminotransferase levels, HIV infection was suspected and then confirmed by a de novo seroconversion of HIV antibody. Subsequently, we noted two consecutive patients with amebic liver abscess, also later found to be infected with HIV. The ameba obtained from these three cases was identified as Entamoeba histolytica by amplification of 16S ribosomal RNA by polymerase chain reaction and direct sequencing. This observation suggests that amebic liver abscess and colitis can be presentations for HIV infection in the Far East. Thus, the local patients with invasive amebiasis, especially those with a protracted course or with risk factors of HIV infection, should be tested for HIV. PMID- 11418796 TI - A Chinese patient with non-HFE-linked iron overload. AB - The gene for hemochromatosis (HFE) was recently identified and contains two missense mutations: C282Y and H63D. The C282Y mutation is found homozygous in approximately 85% to 90% of patients of Northern European ancestry with hereditary hemochromatosis. There are no previous reports with results of genetic testing in Chinese patients with regard to iron overload. In this case report, we describe a Chinese woman with marked hepatic iron overload that was nonfamilial, with unusual biopsy findings, in whom neither the C282Y nor the H63D mutations in HFE were found. PMID- 11418797 TI - Solitary rectal ulcer: a rare cause of gastrointestinal bleeding in an adolescent with hemophilia A. AB - Solitary rectal ulcer syndrome (SRUS) is a rarely reported condition in children. The typical presentation is one of anorectal pain with passage of blood and mucus per rectum in the setting of defecation abnormalities. Diagnosis is made via endoscopy and biopsy. Solitary rectal ulcer syndrome alone is usually a benign condition; however, significant morbidity can occur if complicated by underlying disease states. We report an adolescent with hemophilia A and SRUS who presented with a rectal bleed that required blood transfusion. PMID- 11418798 TI - Boerhaave's syndrome: a pediatric case. AB - BACKGROUND: The term Boerhaave's syndrome refers to esophageal rupture after a sudden increase in intraluminal pressure. This syndrome is most common among alcoholic adult men. This case report is that of a pediatric patient with a similar pathologic process. STUDY: A 6-month-old Hispanic boy with acute lymphoblastic leukemia on chemotherapy arrested 3 weeks into his induction therapy. Diagnostic and therapeutic procedures revealed a hydropneumothorax, which was managed with chest tube drainage. The underlying etiology was revealed 10 days later when feeds were introduced and found exiting the chest tube. A contrast study confirmed esophageal perforation, which was surgically repaired. CONCLUSION: A nontraumatic hydropneumothorax should raise the possibility of Boerhaave's syndrome (spontaneous esophageal rupture), even in pediatric patients. Such a finding warrants a contrast study to facilitate early diagnosis. PMID- 11418799 TI - Synchronous gastric adenocarcinomas in a patient with Meckel's diverticulum. AB - Meckel's diverticulum, which is the most common congenital anomaly of the gastrointestinal tract, occurs when the vitelline duct persists past the 7th week of gestation. Although complications may occur in 8% to 22% of patients with Meckel's diverticula, adenocarcinoma is very uncommon. We describe a patient with early gastric cancer who was incidentally found to have a superficial adenocarcinoma arising from ectopic gastric mucosa within a Meckel's diverticulum. To the best of our knowledge, synchronous gastric adenocarcinoma in a patient with Meckel's diverticulum has not been previously reported. PMID- 11418800 TI - Cerebellar metastasis as a unique presenting feature of gastric cancer. AB - Gastric cancer is often diagnosed in middle-aged patients undergoing upper gastrointestinal endoscopy for abdominal complaints or constitutional symptoms, such as dyspepsia, vomiting or anorexia, weight loss, anemia, jaundice, and ascites. Sometimes, all of these symptoms may be absent, and gastric cancer is diagnosed after detection of metastases to target organs, such as the liver or lung. In a few rare cases, however, even these metastatic localizations may be absent, and clinical signs are only represented by atypical symptoms caused by neurologic metastatic involvement. We report an exceptionally rare case of gastric cancer in which the only presenting symptoms were headache and dizziness caused by a single cerebellar metastasis. PMID- 11418801 TI - Colchicine for persistent constipation after total abdominal colectomy with ileorectostomy for colonic inertia. AB - Severe constipation caused by colonic inertia may be associated with a generalized gastrointestinal dysmotility syndrome. Patients with severe constipation pose a significant challenge in terms of management. Failure of medical therapy usually leads to surgery in the form of a subtotal colectomy. Most patients develop diarrhea after the surgery, but a subgroup of patients continue to experience constipation. We report the case of such a patient who underwent a total abdominal colectomy and ileorectostomy for intractable constipation. He continued to have constipation after the surgery that was responsive only to a gallon of bowel-cleansing solution. A treatment trial of colchicine was partially unsuccessful when used on its own, but a combination of colchicine and a lesser dose of Colyte was found to be effective in maintaining regular bowel movements. PMID- 11418802 TI - Comparison of 5-amino salicylic acid plus glucocorticosteroid with metronidazole and ciprofloxacin in patients with active ulcerative colitis. PMID- 11418803 TI - Acute pancreatitis as the first presentation of ulcerative colitis. PMID- 11418804 TI - There is no significant increase in the incidence of Helicobacter pylori infection in patients with inflammatory bowel disease in Turkey. PMID- 11418805 TI - Infectious mononucleosis presenting with severe cholestatic liver disease in the elderly. PMID- 11418806 TI - Hydatic cyst of spleen. PMID- 11418807 TI - The rising of celiac disease at the sunset of puerperium. PMID- 11418808 TI - Synchronous pancreatic and colon primary cancers. PMID- 11418810 TI - Some advice for the president. PMID- 11418811 TI - Database of molecular probes set to boost chemical genetics. PMID- 11418812 TI - Asthma study death spurs inquiry. PMID- 11418813 TI - Texas facilities count cost of tropical storm. PMID- 11418816 TI - Stem-cell research delayed by German ethics council. PMID- 11418817 TI - Ruling makes it harder to convict dig thieves. PMID- 11418818 TI - Deputies caught on the wrong side of the law. PMID- 11418824 TI - Poetry tells us that our souls have a shadow: can science respond? PMID- 11418823 TI - The search for autism's roots. PMID- 11418825 TI - Call to work together on microarray data analysis. PMID- 11418826 TI - Medical luminaries. PMID- 11418834 TI - Risk and uncertainty. PMID- 11418835 TI - The zombie within. PMID- 11418836 TI - Non-stick water. PMID- 11418837 TI - Structural biology. Chlorophylls galore. PMID- 11418839 TI - Astronomy. Giants in the asteroid belt. PMID- 11418840 TI - Evolutionary biology. Seeing red in speciation. PMID- 11418841 TI - Condensed-matter physics. Nickel probes superconductivity. PMID- 11418842 TI - Neuroscience. Awareness of space. PMID- 11418843 TI - Ecology. Dairy declines hard to swallow. PMID- 11418846 TI - The web of human sexual contacts. PMID- 11418847 TI - Biomechanics. Turning the key on Drosophila audition. PMID- 11418848 TI - Three-dimensional structure of cyanobacterial photosystem I at 2.5 A resolution. AB - Life on Earth depends on photosynthesis, the conversion of light energy from the Sun to chemical energy. In plants, green algae and cyanobacteria, this process is driven by the cooperation of two large protein-cofactor complexes, photosystems I and II, which are located in the thylakoid photosynthetic membranes. The crystal structure of photosystem I from the thermophilic cyanobacterium Synechococcus elongatus described here provides a picture at atomic detail of 12 protein subunits and 127 cofactors comprising 96 chlorophylls, 2 phylloquinones, 3 Fe4S4 clusters, 22 carotenoids, 4 lipids, a putative Ca2+ ion and 201 water molecules. The structural information on the proteins and cofactors and their interactions provides a basis for understanding how the high efficiency of photosystem I in light capturing and electron transfer is achieved. PMID- 11418849 TI - Skyrmions in a ferromagnetic Bose-Einstein condensate. AB - Multi-component Bose-Einstein condensates provide opportunities to explore experimentally the wealth of physics associated with the spin degrees of freedom. The ground-state properties and line-like vortex excitations of these quantum systems have been studied theoretically. In principle, nontrivial spin textures consisting of point-like topological excitations, or skyrmions, could exist in a multi-component Bose-Einstein condensate, owing to the superfluid nature of the gas. Although skyrmion excitations are already known in the context of nuclear physics and the quantum-Hall effect, creating these excitations in an atomic condensate would offer an opportunity to study their physical behaviour in much greater detail, while also enabling an ab initio comparison between theory and experiment. Here we investigate theoretically the stability of skyrmions in a fictitious spin-1/2 condensate of 87Rb atoms. We find that skyrmions can exist in such a gas only as a metastable state, but with a lifetime comparable to (or even longer than) the typical lifetime of the condensate itself. PMID- 11418850 TI - Interplay of magnetism and high-Tc superconductivity at individual Ni impurity atoms in Bi2Sr2CaCu2O8+delta. AB - Magnetic interactions and magnetic impurities are destructive to superconductivity in conventional superconductors. By contrast, in some unconventional macroscopic quantum systems (such as superfluid 3He and superconducting UGe2), the superconductivity (or superfluidity) is actually mediated by magnetic interactions. A magnetic mechanism has also been proposed for high-temperature superconductivity. Within this context, the fact that magnetic Ni impurity atoms have a weaker effect on superconductivity than non magnetic Zn atoms in the high-Tc superconductors has been put forward as evidence supporting a magnetic mechanism. Here we use scanning tunnelling microscopy to determine directly the influence of individual Ni atoms on the local electronic structure of Bi2Sr2CaCu2O8+delta. At each Ni site we observe two d-wave impurity states of apparently opposite spin polarization, whose existence indicates that Ni retains a magnetic moment in the superconducting state. However, analysis of the impurity-state energies shows that quasiparticle scattering at Ni is predominantly non-magnetic. Furthermore, we show that the superconducting energy gap and correlations are unimpaired at Ni. This is in strong contrast to the effects of non-magnetic Zn impurities, which locally destroy superconductivity. These results are consistent with predictions for impurity atom phenomena derived from a magnetic mechanism. PMID- 11418851 TI - Liquid marbles. AB - The transport of a small amount of liquid on a solid is not a simple process, owing to the nature of the contact between the two phases. Setting a liquid droplet in motion requires non-negligible forces (because the contact-angle hysteresis generates a force opposing the motion), and often results in the deposition of liquid behind the drop. Different methods of levitation electrostatic, electromagnetic, acoustic, or even simpler aerodynamic techniques have been proposed to avoid this wetting problem, but all have proved to be rather cumbersome. Here we propose a simple alternative, which consists of encapsulating an aqueous liquid droplet with a hydrophobic powder. The resulting 'liquid marbles' are found to behave like a soft solid, and show dramatically reduced adhesion to a solid surface. As a result, motion can be generated using gravitational, electrical and magnetic fields. Moreover, because the viscous friction associated with motion is very small, we can achieve quick displacements of the droplets without any leaks. All of these features are of potential benefit in microfluidic applications, and also permit the study of a drop in a non wetting situation-an issue of renewed interest following the recent achievement of super-hydrophobic substrates. PMID- 11418852 TI - Decreasing overflow from the Nordic seas into the Atlantic Ocean through the Faroe Bank channel since 1950. AB - The overflow of cold, dense water from the Nordic seas, across the Greenland Scotland ridge and into the Atlantic Ocean is the main source for the deep water of the North Atlantic Ocean. This flow also helps drive the inflow of warm, saline surface water into the Nordic seas. The Faroe Bank channel is the deepest path across the ridge, and the deep flow through this channel accounts for about one-third of the total overflow. Previous work has demonstrated that the overflow has become warmer and less saline over time. Here we show, using direct measurements and historical hydrographic data, that the volume flux of the Faroe Bank channel overflow has also decreased. Estimating the volume flux conservatively, we find a decrease by at least 20 per cent relative to 1950. If this reduction in deep flow from the Nordic seas is not compensated by increased flow from other sources, it implies a weakened global thermohaline circulation and reduced inflow of Atlantic water to the Nordic seas. PMID- 11418853 TI - Metamorphic core complex formation by density inversion and lower-crust extrusion. AB - Metamorphic core complexes are domal uplifts of metamorphic and plutonic rocks bounded by shear zones that separate them from unmetamorphosed cover rocks. Interpretations of how these features form are varied and controversial, and include models involving extension on low-angle normal faults, plutonic intrusions and flexural rotation of initially high-angle normal faults. The D'Entrecasteaux islands of Papua New Guinea are actively forming metamorphic core complexes located within a continental rift that laterally evolves to sea-floor spreading. The continental rifting is recent (since approximately 6 Myr ago), seismogenic and occurring at a rapid rate ( approximately 25 mm yr-1). Here we present evidence-based on isostatic modelling, geological data and heat-flow measurements-that the D'Entrecasteaux core complexes accommodate extension through the vertical extrusion of ductile lower-crust material, driven by a crustal density inversion. Although buoyant extrusion is accentuated in this region by the geological structure present-which consists of dense ophiolite overlaying less-dense continental crust-this mechanism may be generally applicable to regions where thermal expansion lowers crustal density with depth. PMID- 11418854 TI - The elastic constants of MgSiO3 perovskite at pressures and temperatures of the Earth's mantle. AB - The temperature anomalies in the Earth's mantle associated with thermal convection can be inferred from seismic tomography, provided that the elastic properties of mantle minerals are known as a function of temperature at mantle pressures. At present, however, such information is difficult to obtain directly through laboratory experiments. We have therefore taken advantage of recent advances in computer technology, and have performed finite-temperature ab initio molecular dynamics simulations of the elastic properties of MgSiO3 perovskite, the major mineral of the lower mantle, at relevant thermodynamic conditions. When combined with the results from tomographic images of the mantle, our results indicate that the lower mantle is either significantly anelastic or compositionally heterogeneous on large scales. We found the temperature contrast between the coldest and hottest regions of the mantle, at a given depth, to be about 800 K at 1,000 km, 1,500 K at 2,000 km, and possibly over 2,000 K at the core-mantle boundary. PMID- 11418855 TI - Major fungal lineages are derived from lichen symbiotic ancestors. AB - About one-fifth of all known extant fungal species form obligate symbiotic associations with green algae, cyanobacteria or with both photobionts. These symbioses, known as lichens, are one way for fungi to meet their requirement for carbohydrates. Lichens are widely believed to have arisen independently on several occasions, accounting for the high diversity and mixed occurrence of lichenized and non-lichenized (42 and 58%, respectively) fungal species within the Ascomycota. Depending on the taxonomic classification chosen, 15-18 orders of the Ascomycota include lichen-forming taxa, and 8-11 of these orders (representing about 60% of the Ascomycota species) contain both lichenized and non-lichenized species. Here we report a phylogenetic comparative analysis of the Ascomycota, a phylum that includes greater than 98% of known lichenized fungal species. Using a Bayesian phylogenetic tree sampling methodology combined with a statistical model of trait evolution, we take into account uncertainty about the phylogenetic tree and ancestral state reconstructions. Our results show that lichens evolved earlier than believed, and that gains of lichenization have been infrequent during Ascomycota evolution, but have been followed by multiple independent losses of the lichen symbiosis. As a consequence, major Ascomycota lineages of exclusively non-lichen-forming species are derived from lichen forming ancestors. These species include taxa with important benefits and detriments to humans, such as Penicillium and Aspergillus. PMID- 11418856 TI - Phylogenetic analyses do not support horizontal gene transfers from bacteria to vertebrates. AB - Horizontal gene transfer (HGT) has long been recognized as a principal force in the evolution of genomes. Genome sequences of Archaea and Bacteria have revealed the existence of genes whose similarity to loci in distantly related organisms is explained most parsimoniously by HGT events. In most multicellular organisms, such genetic fixation can occur only in the germ line. Therefore, it is notable that the publication of the human genome reports 113 incidents of direct HGT between bacteria and vertebrates, without any apparent occurrence in evolutionary intermediates, that is, non-vertebrate eukaryotes. Phylogenetic analysis arguably provides the most objective approach for determining the occurrence and directionality of HGT. Here we report a phylogenetic analysis of 28 proposed HGT genes, whose presence in the human genome had been confirmed by polymerase chain reaction (PCR). The results indicate that most putative HGT genes are present in more anciently derived eukaryotes (many such sequences available in non vertebrate EST databases) and can be explained in terms of descent through common ancestry. They are, therefore, unlikely to be examples of direct HGT from bacteria to vertebrates. PMID- 11418857 TI - Divergent sexual selection enhances reproductive isolation in sticklebacks. AB - Sexual selection may facilitate speciation because it can cause rapid evolutionary diversification of male mating signals and female preferences. Divergence in these traits can then contribute to reproductive isolation. The sensory drive hypothesis predicts that three mechanisms underlie divergence in sexually selected traits: (1) habitat-specific transmission of male signals; (2) adaptation of female perceptual sensitivity to local ecological conditions; and (3) matching of male signals to female perceptual sensitivity. I test these mechanisms in threespine sticklebacks (Gasterosteus spp.) that live in different light environments. Here I show that female perceptual sensitivity to red light varies with the extent of redshift in the light environment, and contributes to divergent preferences. Male nuptial colour varies with environment and is tuned to female perceptual sensitivity. The extent of divergence among populations in both male signal colour and female preference for red is correlated with the extent of reproductive isolation in these recently diverged species. These results demonstrate that divergent sexual selection generated by sensory drive contributes to speciation. PMID- 11418858 TI - Nodulation of legumes by members of the beta-subclass of Proteobacteria. AB - Members of the Leguminosae form the largest plant family on Earth, with around 18,000 species. The success of legumes can largely be attributed to their ability to form a nitrogen-fixing symbiosis with specific bacteria known as rhizobia, manifested by the development of nodules on the plant roots in which the bacteria fix atmospheric nitrogen, a major contributor to the global nitrogen cycle. Rhizobia described so far belong exclusively to the alpha-subclass of Proteobacteria, where they are distributed in four distinct phylogenetic branches. Although nitrogen-fixing bacteria exist in other proteobacterial subclasses, for example Herbaspirillum and Azoarcus from the phylogenetically distant beta-subclass, none has been found to harbour the nod genes essential for establishing rhizobial symbiosis. Here we report the identification of proteobacteria from the beta-subclass that nodulate legumes. This finding shows that the ability to establish a symbiosis with legumes is more widespread in bacteria than anticipated to date. PMID- 11418859 TI - Spatial awareness is a function of the temporal not the posterior parietal lobe. AB - Our current understanding of spatial behaviour and parietal lobe function is largely based on the belief that spatial neglect in humans (a lack of awareness of space on the side of the body contralateral to a brain injury) is typically associated with lesions of the posterior parietal lobe. However, in monkeys, this disorder is observed after lesions of the superior temporal cortex, a puzzling discrepancy between the species. Here we show that, contrary to the widely accepted view, the superior temporal cortex is the neural substrate of spatial neglect in humans, as it is in monkeys. Unlike the monkey brain, spatial awareness in humans is a function largely confined to the right superior temporal cortex, a location topographically reminiscent of that for language on the left. Hence, the decisive phylogenetic transition from monkey to human brain seems to be a restriction of a formerly bilateral function to the right side, rather than a shift from the temporal to the parietal lobe. One may speculate that this lateralization of spatial awareness parallels the emergence of an elaborate representation for language on the left side. PMID- 11418860 TI - Single neurons in prefrontal cortex encode abstract rules. AB - The ability to abstract principles or rules from direct experience allows behaviour to extend beyond specific circumstances to general situations. For example, we learn the 'rules' for restaurant dining from specific experiences and can then apply them in new restaurants. The use of such rules is thought to depend on the prefrontal cortex (PFC) because its damage often results in difficulty in following rules. Here we explore its neural basis by recording from single neurons in the PFC of monkeys trained to use two abstract rules. They were required to indicate whether two successively presented pictures were the same or different depending on which rule was currently in effect. The monkeys performed this task with new pictures, thus showing that they had learned two general principles that could be applied to stimuli that they had not yet experienced. The most prevalent neuronal activity observed in the PFC reflected the coding of these abstract rules. PMID- 11418861 TI - Bradykinin and nerve growth factor release the capsaicin receptor from PtdIns(4,5)P2-mediated inhibition. AB - Tissue injury generates endogenous factors that heighten our sense of pain by increasing the response of sensory nerve endings to noxious stimuli. Bradykinin and nerve growth factor (NGF) are two such pro-algesic agents that activate G protein-coupled (BK2) and tyrosine kinase (TrkA) receptors, respectively, to stimulate phospholipase C (PLC) signalling pathways in primary afferent neurons. How these actions produce sensitization to physical or chemical stimuli has not been elucidated at the molecular level. Here, we show that bradykinin- or NGF mediated potentiation of thermal sensitivity in vivo requires expression of VR1, a heat-activated ion channel on sensory neurons. Diminution of plasma membrane phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) levels through antibody sequestration or PLC-mediated hydrolysis mimics the potentiating effects of bradykinin or NGF at the cellular level. Moreover, recruitment of PLC-gamma to TrkA is essential for NGF-mediated potentiation of channel activity, and biochemical studies suggest that VR1 associates with this complex. These studies delineate a biochemical mechanism through which bradykinin and NGF produce hypersensitivity and might explain how the activation of PLC signalling systems regulates other members of the TRP channel family. PMID- 11418862 TI - Agonist-independent activation of metabotropic glutamate receptors by the intracellular protein Homer. AB - G-protein-coupled receptors (GPCRs) transduce signals from extracellular transmitters to the inside of the cell by activating G proteins. Mutation and overexpression of these receptors have revealed that they can reach their active state even in the absence of agonist, as a result of a natural shift in the equilibrium between their inactive and active conformations. Such agonist independent (constitutive) activity has been observed for the glutamate GPCRs (the metabotropic glutamate receptors mGluR1a and mGluR5) when they are overexpressed in heterologous cells. Here we show that in neurons, the constitutive activity of these receptors is controlled by Homer proteins, which bind directly to the receptors' carboxy-terminal intracellular domains. Disruption of this interaction by mutagenesis or antisense strategies, or expression of endogenous Homer1a (H1a), induces constitutive activity in mGluR1a or mGluR5. Our results show that these glutamate GPCRs can be directly activated by intracellular proteins as well as by agonists. PMID- 11418863 TI - Nodal signalling in the epiblast patterns the early mouse embryo. AB - Shortly after implantation the mouse embryo comprises three tissue layers. The founder tissue of the embryo proper, the epiblast, forms a radially symmetric cup of epithelial cells that grows in close apposition to the extra-embryonic ectoderm and the visceral endoderm. This simple cylindrical structure exhibits a distinct molecular pattern along its proximal-distal axis. The anterior-posterior axis of the embryo is positioned later by coordinated cell movements that rotate the pre-existing proximal-distal axis. The transforming growth factor-beta family member Nodal is known to be required for formation of the anterior-posterior axis. Here we show that signals from the epiblast are responsible for the initiation of proximal-distal polarity. Nodal acts to promote posterior cell fates in the epiblast and to maintain molecular pattern in the adjacent extra embryonic ectoderm. Both of these functions are independent of Smad2. Moreover, Nodal signals from the epiblast also pattern the visceral endoderm by activating the Smad2-dependent pathway required for specification of anterior identity in overlying epiblast cells. Our experiments show that proximal-distal and subsequent anterior-posterior polarity of the pregastrulation embryo result from reciprocal cell-cell interactions between the epiblast and the two extra embryonic tissues. PMID- 11418864 TI - ATR/ATM-mediated phosphorylation of human Rad17 is required for genotoxic stress responses. AB - Genotoxic stress triggers the activation of checkpoints that delay cell-cycle progression to allow for DNA repair. Studies in fission yeast implicate members of the Rad family of checkpoint proteins, which includes Rad17, Rad1, Rad9 and Hus1, as key early-response elements during the activation of both the DNA damage and replication checkpoints. Here we demonstrate a direct regulatory linkage between the human Rad17 homologue (hRad17) and the checkpoint kinases, ATM and ATR. Treatment of human cells with genotoxic agents induced ATM/ATR-dependent phosphorylation of hRad17 at Ser 635 and Ser 645. Overexpression of a hRad17 mutant (hRad17AA) bearing Ala substitutions at both phosphorylation sites abrogated the DNA-damage-induced G2 checkpoint, and sensitized human fibroblasts to genotoxic stress. In contrast to wild-type hRad17, the hRad17AA mutant showed no ionizing-radiation-inducible association with hRad1, a component of the hRad1 hRad9-hHus1 checkpoint complex. These findings demonstrate that ATR/ATM-dependent phosphorylation of hRad17 is a critical early event during checkpoint signalling in DNA-damaged cells. PMID- 11418869 TI - Chimerism testing after allogeneic stem cell transplantation: importance of timing and optimal technique for testing in different clinical-biological situations. PMID- 11418870 TI - Quantitative analysis of chimerism after allogeneic stem cell transplantation using multiplex PCR amplification of short tandem repeat markers and fluorescence detection. PMID- 11418871 TI - Summary of the Guidelines for Preventing Opportunistic Infections among Hematopoietic Stem Cell Transplant Recipients. AB - This article contains highlights of "Guidelines for Preventing Opportunistic Infections among Hematopoietic Stem Cell Transplant Recipients: Recommendations of the CDC, the Infectious Diseases Society of America, and the American Society of Blood and Marrow Transplantation," which was published in the Morbidity and Mortality Weekly Report. There are sections on the prevention of bacterial, viral, fungal, protozoal, and helminth infections and on hospital infection control, strategies for safe living following transplantation, immunizations, and hematopoietic stem cell safety. The guidelines are evidence-based, and prevention strategies are rated by both the strength of the recommendation and the quality of evidence that supports it. Recommendations are given for preventing cytomegalovirus disease with prophylactic or preemptive gancyclovir, herpes simplex virus disease with prophylactic acyclovir, candidiasis with fluconazole, and Pneumocystis carinii pneumonia with trimethoprim-sulfamethoxazole. Hopefully, following the recommendations made in the guidelines will reduce morbidity and mortality from opportunistic infections in hematopoietic stem cell transplant recipients. PMID- 11418872 TI - Utility of semiquantitative polymerase chain reaction for Epstein-Barr virus to measure virus load in pediatric organ transplant recipients with and without posttransplant lymphoproliferative disease. AB - We examined the utility of Epstein-Barr virus (EBV) load as a test for the presence of posttransplant lymphoproliferative disease (PTLD). A semiquantitative (SQ) EBV polymerase chain reaction (PCR) on peripheral blood mononuclear cells (PBMC) was used to determine virus load. We compared the values from pediatric patients, both with and without PTLD, with those from healthy pediatric and adult subjects. The virus loads for asymptomatic healthy subjects had a range of 0-1 log10 cells/10(6) PBMCs. Among transplant recipients (n=135), the mean virus load (+/- standard deviation) at the time of diagnosis of PTLD was 3.1+/-1.2 log(10) cells/10(6) PBMCs versus a baseline value of 1.3+/-1.4 log(10) cells/10(6) PBMCs in children without PTLD (P<.0001). A cutoff of > or =3 log10 cells/10(6) peripheral blood leukocytes resulted in the following values for use of virus load as a test for PTLD: sensitivity, 69%; specificity, 76%; positive predictive value, 28%; and negative predictive value, 95%. We conclude that determination of EBV load by use of SQ PCR is more useful in ruling out than in indicating the presence of PTLD. PMID- 11418873 TI - Acquisition of vancomycin-resistant enterococci during scheduled antimicrobial rotation in an intensive care unit. AB - Scheduled rotation of treatment of gram-negative antimicrobial agents has been associated with reduction of serious gram-negative infections. The impact of this practice on other nosocomial infections has not been assessed. The purpose of this study was to determine if scheduled antimicrobial rotation reduced rates of acquisition of enteric vancomycin-resistant enterococci (VRE) among 740 patients admitted to an intensive care unit (ICU). The preferred gram-negative agent was ceftazidime during rotation 1 and ciprofloxacin during rotation 2. Unadjusted VRE acquisition rates were 8.5 cases per 1000 ICU days and 11.7 cases per 1000 ICU days during rotations 1 and 2, respectively (P<.01). However, scheduled antimicrobial rotation of ceftazidime with ciprofloxacin had no effect on the risk of acquiring VRE in the ICU after adjustment for known risk factors. Independent predictors of acquisition of VRE were enteral feedings, higher colonization pressure, and increased duration of anaerobic therapy. Our findings can confirm no additional beneficial or adverse effect on VRE acquisition among ICU patients as a result of this practice. PMID- 11418874 TI - Etiology, reasons for hospitalization, risk classes, and outcomes of community acquired pneumonia in patients hospitalized on the basis of conventional admission criteria. AB - We performed an observational analysis of prospectively collected data on 533 nonseverely immunosuppressed adult patients who were hospitalized for community acquired pneumonia on the basis of conventional admission criteria. For this population, we correlated etiology, reasons for admission, and outcomes using the Pneumonia Severity Index (PSI), to identify major discrepancies between the PSI risk class and the conventional criteria for deciding the site of care. PSI classes and corresponding mortality rates were as follows: class I, 51 patients (0%); class II, 62 (2%); class III, 117 (3%); class IV, 198 (10%); and class V, 105 (29%). We identified significant discrepancies between both methods. Overall, 230 patients (40%) who were hospitalized according to conventional criteria were assigned to low-risk classes. Of these 230 patients, 137 (60%) needed supplementary oxygen or had pleural complications; for the remaining patients, there were no irrefutable reasons for admission. This latter group deserves prospective evaluation in randomized studies that compare ambulatory and in hospital management. PMID- 11418875 TI - Ultrasonographic detection and assessment of preclinical oesophagostomum bifurcum induced colonic pathology. AB - In northern Ghana and Togo, Oesophagostomum bifurcum infects an estimated 250,000 people, as determined by cultures of stool samples. The juvenile stages of the helminth develop within colonic wall nodules, causing Dapaong tumor or multinodular disease, at the rate of 1 case per week at Nalerigu Hospital in Ghana. Our aim was to discover whether suspected colonic-wall pathology is ultrasonographically visible in asymptomatic individuals living in the area where O. bifurcum is endemic. A total of 464 persons from 3 villages, ranging from highly infected to noninfected, were examined with ultrasonography. Anechogenic colonic lesions with posterior wall enhancement were observed in 71 (54.2%) of 131 and 57 (24.5%) of 233 persons from the villages of endemicity, and no lesions were seen in persons from the village outside the area of endemicity. We describe the lesions noted in this study as nodules caused by O. bifurcum, on the basis of their association at a population level with prevalence of larvae in stools, their expected ultrasonographic appearance and distribution (on the basis of our surgical experience with oesophagostomiasis), and the lack of a convincing differential diagnosis. PMID- 11418876 TI - Treating cardiovascular disease with antimicrobial agents: a survey of knowledge, attitudes, and practices among physicians in the United States. AB - To assess physicians' knowledge, attitudes, and prescribing behaviors with regard to the association between Chlamydia pneumoniae and cardiovascular disease, we surveyed 750 physicians in Alaska, 1172 in West Virginia, and 569 infectious disease (ID) specialists in a nationwide network during February-May 1999. Eighty five percent knew of the association between C. pneumoniae and atherosclerosis, but this awareness was more common among ID specialists and cardiologists than among generalists (96% vs. 77%; P<.001). Knowledge scores were significantly higher among ID specialists and cardiologists (P<.001) and among physicians who saw relatively more patients who had myocardial infarction and/or were at risk of atherosclerotic disease. Four percent of physicians had treated or recommended treating cardiovascular diseases with antimicrobial agents; this percentage was significantly higher among cardiologists, physicians who empirically treat patients with peptic ulcers with antimicrobial agents, and physicians with a relatively high knowledge score. PMID- 11418877 TI - Risk factors for candidal bloodstream infections in surgical intensive care unit patients: the NEMIS prospective multicenter study. The National Epidemiology of Mycosis Survey. AB - To assess risk factors for development of candidal blood stream infections (CBSIs), a prospective cohort study was performed at 6 sites that involved all patients admitted to the surgical intensive care unit (SICU) for >48 h over a 2 year period. Among 4276 such patients, 42 CBSIs occurred (9.82 CBSIs per 1000 admissions). The overall incidence was 0.98 CBSIs per 1000 patient days and 1.42 per 1000 SICU days with a central venous catheter in place. In multivariate analysis, factors independently associated with increased risk of CBSI included prior surgery (relative risk [RR], 7.3), acute renal failure (RR, 4.2), receipt of parenteral nutrition (RR, 3.6), and, for patients who had undergone surgery, presence of a triple lumen catheter (RR, 5.4). Receipt of an antifungal agent was associated with decreased risk (RR, 0.3). Prospective clinical studies are needed to identify which antifungal agents are most protective and which high-risk patients will benefit from antifungal prophylaxis. PMID- 11418878 TI - Invasive candidiasis: turning risk into a practical prevention policy? PMID- 11418879 TI - Bk virus: a clinical review. AB - We present a review of the clinically oriented literature about BK virus, a relative of JC virus, which is the etiologic agent of progressive multifocal leukoencephalopathy (PML). The kidney, lung, eye, liver, and brain have been proposed as sites of BK virus-associated disease, both primary and reactivated. BK virus has also been detected in tissue specimens from a variety of neoplasms. We believe that BK virus is most often permissively present in sites of disease in immunosuppressed patients, rather than being an etiologic agent that causes symptoms or pathologic findings. There is, however, strong evidence for BK virus associated hemorrhagic cystitis and nephritis, especially in recipients of solid organ or bone marrow transplants. Now that BK virus can be identified by use of specific and sensitive techniques, careful evaluation of the clinical and pathologic presentations of patients with BK virus will allow us to form a clearer picture of viral-associated pathophysiology in many organ systems. PMID- 11418880 TI - Patient selection criteria and management guidelines for outpatient parenteral antibiotic therapy for native valve infective endocarditis. AB - Outpatient parenteral antibiotic therapy (OPAT) for infective endocarditis (IE) is being applied widely, despite the absence of controlled data that demonstrates that outcomes are equivalent to those with standard inpatient antibiotic therapy. We review existing OPAT guidelines, published data on the timing of complications from IE, and data on risk factors that can be used to predict complications. These data are used to propose more stringent criteria for patient selection and clinical management of OPAT for native valve IE. We recommend a conservative approach (inpatient or daily outpatient follow-up) during the critical phase (weeks 0-2 of treatment), when complications are most likely, and we recommend consideration of OPAT for the continuation phase (weeks 2-4 or 2-6 of treatment) when life-threatening complications are less likely. PMID- 11418881 TI - Vancomycin-resistant enterococci: mechanisms and clinical observations. AB - Enterococci are not generally regarded as highly virulent bacterial pathogens. However, resistance to many antimicrobial drugs complicates treatment of enterococcal infections. Acquired resistance to high concentrations of glycopeptide antibiotics, specifically vancomycin, has exacerbated this problem. This article seeks to concisely review the mechanisms of that resistance and its effects on clinical management of enterococcal infections, as well as clinical microbiology and infection control. PMID- 11418882 TI - Rapid identification of bacteria and yeast: summary of a National Committee for Clinical Laboratory Standards proposed guideline. AB - Using colony morphology on selected agars, Gram-stain morphology, and a number of 1-step biochemical or enzymatic tests, skilled microbiologists can identify the species of the majority of isolates seen routinely in a clinical laboratory. These results are often available more quickly than and are as accurate as those derived from conventional methods. The National Committee for Clinical Laboratory Standards has produced a guideline that describes tests that can be used to identify a number of aerobic gram-negative rods and gram-positive cocci, a number of commonly isolated anaerobes, and 3 species of yeast. An overview of the organisms included in the guideline, the tests that identify them, and the situations in which rapid testing is appropriate is presented here. PMID- 11418883 TI - Malaria chemoprophylaxis in the age of drug resistance. I. Currently recommended drug regimens. AB - As international travel becomes increasingly common and resistance to antimalarial drugs escalates, a growing number of travelers are at risk for contracting malaria. Parasite resistance to chloroquine and proguanil and real or perceived intolerance among patients to standard prophylactic agents such as mefloquine have highlighted the need for new antimalarial drugs. Promising new regimens include atovaquone and proguanil, in combination; primaquine; and a related 8-aminoquinoline, tafenoquine. These agents are active against the liver stage of the malaria parasite and therefore can be discontinued shortly after the traveler leaves an area where malaria is endemic, which encourages adherence to the treatment regimen. Part 1 of this series reviews currently recommended chemoprophylactic drug regimens, and part 2 will focus on 8-aminoquinoline drugs. PMID- 11418884 TI - Disseminated nocardiosis in a patient with X-linked chronic granulomatous disease and human immunodeficiency virus infection. AB - We report the first case of HIV infection in a patient with underlying X-linked chronic granulomatous disease (CGD) who presented with hepatopulmonary nocardiosis. Despite the coexistence of CGD and HIV, the response to therapy was normal, and no unusual sequelae were noted. The patient's high virus burden was successfully repressed with antiretroviral therapy, suggesting that the nicotinamide adenine dinucleotide phosphate oxidase system is not essential for active viral replication or response to antiretroviral agents. PMID- 11418885 TI - Hepatitis C virus infection-related morbidity and mortality among patients with human immunodeficiency virus infection. AB - Hepatitis C virus (HCV) has emerged as a major pathogen among patients with human immunodeficiency virus (HIV). Morbidity and mortality were compared among 263 patients with HIV alone, 166 patients with HIV and HCV, and 60 patients with HCV alone (mean duration of follow-up, 2 years and 10 months). No differences in HIV loads and CD4 cells counts were observed between the HIV and HIV/HCV groups. Alanine aminotransferase levels were higher (52 U/L versus 35 U/L; P<.05) and albumin levels were lower (3.5 g/dL versus 3.8 g/dL; P <.02) among coinfected patients than they were among patients with HIV alone. Liver decompensation developed in 10% of patients with HIV/HCV coinfection. In contrast, no liver related deaths or decompensation occurred in patients without coinfection (P<.05). Of the patients with HIV alone, 7% died, compared with 11% of the coinfected patients (P<.02); 47% of the deaths in the latter group were due to liver-related causes. In summary, HCV infection causes increased morbidity and mortality in patients with HIV infection. PMID- 11418886 TI - Trends in bloodstream infections among human immunodeficiency virus-infected adults admitted to a hospital in Nairobi, Kenya, during the last decade. AB - Bloodstream infections are a frequent complication in human immunodeficiency virus (HIV)-infected adults in Africa and usually associated with a poor prognosis. We evaluated bloodstream infections across a decade in 3 prospective cross-sectional surveys of consecutive medical admissions to the Kenyatta National Hospital, Nairobi, Kenya. Participants received standard clinical care throughout. In 1988-1989, 29.5% (28 of 95) of HIV-positive patients had bloodstream infections, compared with 31.9% (46 of 144) in 1992 and 21.3% (43 of 197) in 1997. Bacteremia and mycobacteremia were significantly associated with HIV infection. Infections with Mycobacterium tuberculosis, non-typhi species of Salmonella (NTS), and Streptococcus pneumoniae predominated. Fungemia exclusively due to Cryptococcus neoformans was uncommon. Clinical features at presentation remained similar. Significant improvements in the survival rate were recorded among patients with NTS bacteremia (20%-83%; P<.01) and mycobacteremia (0%-73%; P<.01). Standard clinical management can improve outcomes in resource-poor settings. PMID- 11418887 TI - Recurrent Listeria monocytogenes infection: relapse or reinfection with a unique strain confirmed by molecular subtyping. AB - We report a case of recurrent listeriosis for which molecular subtyping by automated ribotyping and pulsed-field gel electrophoresis confirmed either relapse of infection or reinfection due to a common source almost 9 months after initial infection due to a unique Listeria monocytogenes strain in a patient with colorectal cancer. This case report illustrates the potential use of molecular subtyping to further understand the pathogenesis and epidemiology of listeriosis and the potential for relapse of Listeria infections in humans. PMID- 11418888 TI - Adenovirus type 3 viremia in an adult with toxic shock-like syndrome. AB - Surveillance by the Unexplained Deaths and Critical Illnesses Project (UNEX) uncovered a novel presentation of adenovirus type 3 infection that satisfied the criteria for toxic shock-like syndrome in a 28-year-old immunocompetent man. Adenovirus may be a cause of toxic shock syndrome; surveillance systems such as UNEX may uncover additional causes of this and other clinically defined infectious syndromes. PMID- 11418889 TI - Predominant kidney involvement in a fatal case of hantavirus pulmonary syndrome caused by Sin Nombre virus. AB - A 27-year-old woman presented to a hospital with symptoms resembling pyelonephritis; respiratory distress did not develop until nearly a day after admission and she subsequently died. The Unexplained Deaths and Critical Illnesses Project of the Centers for Disease Control and Prevention confirmed Sin Nombre virus infection by the results of serological testing and sequencing of the viral genome; staining of Sin Nombre virus antigen in the pulmonary capillaries was relatively weak. PMID- 11418890 TI - Disease surveillance versus viral surveillance. PMID- 11418891 TI - Some healthy skepticism about inhaled therapy for tuberculosis. PMID- 11418893 TI - Ciprofloxacin for treatment of tularemia. PMID- 11418894 TI - Osteonecrosis in human immunodeficiency virus-infected patients may not be related to immune reconstitution. PMID- 11418896 TI - Manic syndrome associated with efavirenz overdose. PMID- 11418898 TI - The comfort zone and the fun zone: a case for volunteerism in hand surgery. AB - Hand surgeons have a rich heritage of stewardship to the community through volunteer medical work. There is a wide breadth of service opportunities that can be accomplished at home and overseas. The challenges of volunteerism are balanced by the personal and professional rewards. PMID- 11418899 TI - Effect of absorbable polydioxanone flexor tendon repair and restricted active mobilization in a canine model. AB - We performed an experimental study to evaluate the mechanical and histologic healing process of flexor tendon repair using absorbable polydioxanone monofilament followed by active mobilization. Sixty-four canine flexor digitorum profundus tendons were repaired using polydioxanone monofilament or control braided polyester. Animals were evaluated 7, 14, 28, and 42 days after surgery with 12 day 0 control tendons. All repaired tendons healed without rupture or gap formation. Mechanically, all specimens had adequate tensile strength enabling active mobilization. The tendons that were repaired using braided polyester healed without initial tensile depression; however, the ultimate tensile strength values of polydioxanone specimens showed a significant decrease at day 14. Histologically, an inflammatory response was observed around the polydioxanone monofilament; this reaction increased from day 14 to day 42. However, the inflammatory response did not cause large adhesions or large tendon callus formation. This method of using absorbable suture material for tendon repair has the potential for use in the clinical setting. PMID- 11418900 TI - The role of midcarpal arthroscopy in the diagnosis of disorders of the wrist. AB - The findings of midcarpal versus radiocarpal arthroscopic examinations were compared in the diagnosis of a variety of wrist pathology in 89 patients. During 15 months 89 midcarpal arthroscopic examinations were performed in conjunction with radiocarpal arthroscopic examinations. Eighty-one wrists underwent arthroscopy for acute or chronic intracarpal instability. Eight wrists underwent arthroscopy for arthroscopy-assisted intra-articular distal radius fracture reduction. In the acute wrist instability group midcarpal arthroscopy added to the radiocarpal diagnosis in 21 of 26 (82%) of the wrists. In the chronic wrist instability group midcarpal arthroscopy added to the radiocarpal diagnosis in 46 of 55 (84%) of the wrists. In the distal radius group 5 of 8 wrists had additional pathology on the midcarpal arthroscopy examination, leading to additional surgical intervention. These results demonstrate that midcarpal arthroscopy added statistically significant information to the radiocarpal examination compared with wrist arthroscopy performed without a midcarpal examination. PMID- 11418901 TI - Computed tomography scanning of intra-articular distal radius fractures: does it influence treatment? AB - The purpose of this study is to determine whether the addition of computed tomography (CT) results in changes in the evaluation and treatment of intra articular distal radius fractures. Fifteen intra-articular distal radius fractures were evaluated independently by 4 hand surgeons. Plain x-rays were reviewed initially followed by the corresponding CT scans for comparison of articular step-off and gapping, comminution, and treatment. Kappa coefficients (kappa) of intraobserver and interobserver reliability for treatment plans were generated. Computed tomography scans improved the sensitivity of measurement of articular surface gapping, improved the accuracy of detection of comminution and distal radioulnar joint involvement, and altered proposed treatment plans within observers (intraobserver agreement: kappa =.54, moderate) and improved agreement of proposed treatment plans between observers (kappa =.34 to kappa =.44, fair to moderate). Computed tomography scanning influenced observers to change treatment plans and resulted in increased interobserver reliability in the proposed management of these injuries. PMID- 11418902 TI - Lunotriquetral arthrodesis using cancellous bone graft. AB - Previous reports of lunotriquetral arthrodesis suggest relatively low rates of primary fusion without the use of permanent fixation and/or prolonged immobilization. We performed 26 lunotriquetral arthrodeses in 24 patients with a technique using cancellous bone graft to fill a biconcave space created in the adjoining bones with parallel K-wire fixation. The indications included symptomatic lunotriquetral instability and degenerative arthritis. All patients received conservative treatment before surgery. Primary fusion was achieved in all wrists in an average of 50 days. Postoperative wrist flexion/extension averaged 77%/80% of unaffected sides. Radial/ulnar deviation averaged 95%/91% of unaffected sides. Pain relief was good or very good in 83%. Eight-eight percent returned to the workforce. This study shows reliable, effective results with a technique that adheres to the principles of intercarpal arthrodesis and avoids permanent internal fixation. PMID- 11418903 TI - Type I versus type II lunates: Ligament anatomy and presence of arthrosis. AB - One hundred-seventy embalmed cadaver wrists were dissected. The type of lunate (type I, no medial hamate facet; type II, medial hamate facet), the incidence and location of arthrosis (exposed subchondral bone) in the lunohamate joint, and the anatomic relationship of the volar triquetrocapitate (T-C) and the volar triquetrohamate (T-H) ligaments were identified and measured. The relationship between the T-C and T-H ligaments was classified into 3 types. In type A the T-C ligament was completely separate from the T-H ligament, in type B the T-C ligament overlapped the T-H ligament, and in type C the T-C ligament had an additional ligament from the triquetrum to the proximal pole of the hamate. Eighty-two percent of type I lunates were associated with a type A relationship between the T-C and the T-H ligaments and 96% of type II lunates were associated with a type C relationship between the T-C and the T-H ligaments. Arthrosis at the proximal pole of the hamate was more commonly associated with the type II lunates (49%) and with the type C relationship (57%) of the T-C and T-H ligaments. The incidence of arthrosis in the lunohamate joint was also significantly greater in the type II lunate with a medial facet of 3 mm or more. PMID- 11418904 TI - Vascularized bone graft from the index metacarpal for Kienbock's disease: a case report. AB - A vascularized bone graft from the head and neck of the index metacarpal bone was used for the treatment of stage IIIA Kienbock's disease in a 20-year-old woman. Three months after surgery pain had disappeared and wrist range of motion improved. Postoperative radiographs and magnetic resonance imaging revealed revascularization and fracture healing of the lunate bone. This technique has not been used in the treatment of Kienbock's disease; it may be useful in selected cases. PMID- 11418905 TI - Subcutaneous herniation of tendon interposition after trapeziectomy in three cases: explanation and implications. AB - We report a complication following trapeziectomy and tendon interposition. Subcutaneous herniation of the tendon interposition occurred in 3 of 412 cases in which trapeziectomy, ligament reconstruction, and tendon interposition were performed. This herniation occurred posterolaterally in the early postoperative period and resulted in dorsal swelling and superficial pain. Magnetic resonance imaging was helpful in confirming the diagnosis and excision of the herniated interposition material resulted in satisfactory pain relief and functional outcome as long as metacarpal stability was present. PMID- 11418906 TI - Radial artery pseudoaneurysm complication from use of AO/ASIF volar distal radius plate: a case report. AB - A case of pseudoaneurysm of the radial artery after fixation of a distal radius using the AO/ASIF volar distal radius plate system is presented. This unusual complication resulted from inadequate fracture stabilization and plate preparation. Technical tips on plate shortening and preparation of the cut ends are emphasized. PMID- 11418907 TI - Sigmoid notch osteoplasty for chronic volar instability of the distal radioulnar joint: a case report. AB - Chronic recurrent volar dislocation of the distal radioulnar joint is a rare condition for which many operative procedures have been described. We describe a case of painful dislocation on supination. Investigation of the distal radioulnar joint revealed the absence of degenerative arthritis and a flat sigmoid notch that permitted dislocation. The case was successfully treated by osteoplasty of the volar lip of the sigmoid notch to create a buttress that prevented volar dislocation. PMID- 11418908 TI - The prevalence and characteristics of nerve compression symptoms in the general population. AB - The purpose of this cross-sectional study was to evaluate the prevalence and intensity of nerve compression symptoms and to estimate the prevalence of carpal tunnel syndrome (CTS) in the general population. A survey that included the Katz hand diagram, the Carpal Tunnel Instrument (CTI), and the Short Form-36 questionnaire was sent to 1,559 people. A short telephone survey was conducted to a random sample of 110 nonresponders to determine if they were systematically different from the responders. Of the responders 35.1% had a symptom severity (CTI subscale) score of > or =1.5. Of the responders and the nonresponders 23.2% and 14.5%, respectively, reported waking at least once per night with numbness; 37.3% of the responders and 33.6% of the nonresponders experienced pain in the hand at least once per day. As determined by the Katz hand diagrams, 58 (16.3%) of the responders had classic or probable distributions of symptoms (likely to have CTS) and 298 (83.7%) had possible and unlikely distributions. After correcting for nonresponders our lowest possible estimate of CTS prevalence in the general US population is 3.72%, indicating a larger pool of symptomatic people than previously reported. PMID- 11418909 TI - Congenital constriction band syndrome causing ulnar nerve palsy: early diagnosis and surgical release with long-term follow-up. AB - Three children with congenital constriction band syndrome affecting their upper extremities demonstrated clinical and electrophysiologic signs of a complete ulnar nerve palsy. Two of the children were diagnosed immediately postpartum with the subtle findings of an intrinsic minus posture of their hand and inability to actively extend their fingers at the proximal interphalangeal joints. One child had at least 5.5 months of intrauterine compression of the ulnar nerve detected by ultrasound examination at 18 weeks. Despite early release of the constriction bands, at 3 months in 2 children and at 6 months in 1 child, the ulnar nerve palsies persisted for a mean follow-up period of 7 years. If clinical examination of an infant with constriction band syndrome is indicative of a complete ulnar nerve palsy, the constriction band should be released as early as possible. If surgical exploration reveals significant compression of the ulnar nerve, consideration should be given to excising the involved segment of nerve with immediate primary nerve repair or nerve grafting because even early release of the constriction band does not seem to result in neurologic improvement in long term follow-up studies. PMID- 11418910 TI - Neuropathy of motor branch of median or ulnar nerve induced by midpalm ganglion. AB - Two cases of neuropathy of a motor branch caused by a midpalmal ganglion are presented. In the first case the ganglion originated from the midcarpal joint, protruded into the thenar muscle, and compressed the motor branch of the median nerve. In the second case the ganglion, distal to the fibrous arch of the hypothenar muscles, originated from the third carpometacarpal joint and compressed the motor branch of the ulnar nerve. In both cases muscle weakness and finger deformity recovered well after resection of the ganglion. This clinical condition is rare compared with carpal tunnel syndrome and Guyon's tunnel syndrome, which are caused by a ganglion in the wrist. PMID- 11418911 TI - Axonal regeneration stimulated by the combination of nerve growth factor and ciliary neurotrophic factor in an end-to-side model. AB - The aim of this study was to investigate the potential for stimulating axonal regeneration in the context of end-to-side coaptation using a combination of nerve growth factor and ciliary neurotrophic factor in the rat sciatic nerve model. Four experimental groups (n = 8) were used: end-to-side coaptation only, end-to-side coaptation plus growth factor injection, primary repair, and nontransferred gap control. Twenty weeks after surgery histologic analysis showed that the ratio of axon density was significantly increased for the growth factor injection group. Histologic evidence suggested contamination from the proximal peroneal stump. Electrical stimulation and muscle weights showed that the target muscles had been reinnervated in all groups except the nontransferred gap control group. These data support the conclusion that the use of nerve growth factor and ciliary neurotrophic factor in combination may enhance regeneration in the peripheral nervous system. This is consistent with previous reports on the central nervous system and suggests a potential application in future studies aimed at improving peripheral nerve regeneration. Another conclusion is that contamination from the proximal peroneal stump may explain the regeneration observed in the end-to-side model. Further study using retrograde labeling is needed to establish the origin of the regenerating axons. Finally, evidence suggests that regenerating axons can use the epineurium of an intact nerve to bridge a gap in continuity. PMID- 11418912 TI - Bone suture anchors versus the pullout button for repair of distal profundus tendon injuries: a comparison of strength in human cadaveric hands. AB - Avulsion or distal tendon laceration of flexor digitorum profundus (FDP) is classically repaired to the base of the distal phalanx via a pullout suture over a button. Bone suture anchors, used extensively in other surgical areas, have recently been proposed for reattachment of the FDP to the distal phalanx. The FDP tendons of the index, long, and ring fingers in 9 fresh frozen cadeveric hands were randomized to 1 of 3 repair techniques after simulated distal avulsion injuries. These were the pullout button using 3-0 monofilament nylon in a 2 strand Bunnell suture pattern, the 1.8 mm Mini QuickAnchor (Mitek Products, Norwood, MA) using 3-0 braided polyester in a 2-strand Bunnell suture pattern, and the Mitek micro anchor using 3-0 braided polyester with a modified 4-strand Becker suture pattern. Nine specimens were loaded to failure, noting maximum load and mode of failure. The 1.3 mm Micro QuickAnchor (Mitek) technique (69.6 +/- 10.8 N) was significantly stronger than the pullout button (43.3 +/- 4.8 N) or the Mini anchor technique (44.6 +/- 12.7 N). The Micro bone suture anchor provides a stronger tendon to bone repair than the pullout button or the Mini anchor. Given the disadvantages of the pullout button, the Micro bone suture anchor with the modified Becker technique is worth consideration as an alternative method to repair distal FDP avulsions. PMID- 11418913 TI - Prospective randomized trial of open versus percutaneous surgery for trigger digits. AB - Various methods for the treatment of trigger digits exist. This study was designed to compare the results of an open surgical technique with those of a percutaneous surgical technique for the treatment of trigger digits. Ninety-six patients with 100 trigger digits were randomized to either open (n = 46) or percutaneous (n = 54) surgical release of the first annular pulley. Operation time, duration of postoperative pain, recovery of motor function, and surgical complications were assessed. Trigger digits were successfully treated in 98% of the cases using the open surgical technique and in 100% of the cases using the percutaneous technique. Mean operation time was significantly longer using the open technique. Mean duration of postoperative pain and time to recovery of motor function were significantly shorter for patients treated with the percutaneous method. No serious complications were observed in either group. We conclude that percutaneous correction of trigger digits is a quicker procedure, is less painful, and shows significantly better results in rehabilitation than open surgery. PMID- 11418914 TI - Congenital absence of the juncturae tendini contributing to dislocation of the extensor tendons. AB - We report a patient with bilateral congenital absence of the juncturae tendini who developed dislocation of the extensor tendons of the long and ring fingers of both hands. Reconstruction of the juncturae tendini restored extensor function. We describe our technique and the results 9 months after surgery. PMID- 11418915 TI - The distally based radial forearm fascia-fat flap for treatment of recurrent de Quervain's tendonitis. AB - A radial fascial flap has been described as a valuable and versatile option to provide appropriate tissue coverage of tendons, nerves, and soft tissues in the forearm and hand. We report the use of this distally based radial forearm fascia fat flap to create a fascial tube to treat recurrent de Quervain's tendonitis. PMID- 11418916 TI - In vivo determination of available brachioradialis excursion during tetraplegia reconstruction. AB - The brachioradialis muscle is a common donor in tetraplegia reconstruction. The purpose of this study was to determine the available excursion of the brachioradialis during graduated release of its insertion sites. Fourteen extremities in 10 patients with cervical spine injuries underwent surgical reconstruction for restoration of hand function using the brachioradialis as a donor for one of the restored movements. At the time of surgery the brachioradialis was exposed along the forearm and a wire was inserted into the muscle followed by excitation with a biphasic, asymmetrical, charge-balanced waveform. A 12-Hz frequency and 20-mA current were used to stimulate the muscle while the pulse duration was varied between 0 and 200 milliseconds to achieve maximum contraction. Average brachioradialis excursion after incision of the radial styloid insertion site was 8 mm and 14 mm after mobilization to the musculotendinous junction. Further release of the fascial connections and mobilization of the muscle belly increased the excursion to an average of 61 mm. The increase in excursion after fascial release and muscle mobilization was significant and should be performed to obtain maximum available excursion. PMID- 11418917 TI - A congenital hand deformity: Dupuytren's disease. AB - A 10-month-old child presented with a lack of extension at the distal interphalangeal joint. Despite the absence of trauma, a provisional diagnosis of mallet finger led to treatment using a short dorsal splint. Four months later the flexion had increased and included the proximal interphalangeal joint. Palpation revealed a palmar cord on the lateral aspect of the finger. Surgery disclosed a typical Dupuytren cord and the histology supported this diagnosis. There was no known family history of the disease. There was no sign of recurrence 27 months after surgery in this case of Dupuytren's disease that was present at birth. PMID- 11418918 TI - One-stage elongation of the third or fourth brachymetacarpia through the palmar approach. AB - The technique of 1-stage elongation of a short third or fourth metacarpal through the palmar approach was devised to avoid the drawbacks of the conventional procedures through the dorsal approach with or without using an external fixator. This technique was used in the treatment of 3 cases and the results and the technique are described. PMID- 11418919 TI - Postoperative monitoring of pediatric toe-to-hand transfers with differential pulse oximetry. AB - Twenty-three toe-to-hand transfers performed in 21 children were monitored after surgery using differential pulse oximetry for 4 to 12 days. Pulse rate recorded by the pulse oximetry monitored the patency of the arterial anastomosis, whereas oxygen saturation (SaO(2)) corresponded to the patency of the venous anastomosis; these were both compared with the systemic pulse rate and oxygen saturation recorded by a second control pulse oximeter probe attached to a contralateral finger or toe. All 23 toe transfers were ultimately successful, but 2 required re exploration for anastomotic problems detected by the pulse oximeter. Based on this experience, the following criteria have been developed for the nursing and junior medical staff: if the arterial pulse of the toe transfer is lost completely or if the pulse rate differs significantly from the systemic pulse rate, measured by the second control pulse oximeter, thrombosis of the arterial anastomosis should be suspected. If the oxygen saturation of the toe transfer decreases below the oxygen saturation measured by the control pulse oximeter and this differential is sustained over a period of time, thrombosis of the venous anastomosis should be suspected. Differential pulse oximetry appears to be superior to temperature monitoring and percutaneous and laser Doppler monitoring and provides the most simple and continuous technique of noninvasive postoperative monitoring of toe-to-hand transfers in children requiring reconstruction of traumatic or congenital deformities. PMID- 11418920 TI - Comparison of continuous and interrupted suture techniques in microvascular anastomosis. AB - We evaluated the efficacy of the continuous suture technique (CST) in arteries and veins with varying external diameters (ED). In study 1 a direct end-to-end anastomosis was performed in 5 groups of animals (n = 15 in each group): group 1, rabbit carotid artery (ED, 1.8-2.0 mm); group 2, rabbit femoral artery (ED, 1.4 1.6 mm); group 3, rat femoral artery (ED, 0.7-0.9 mm); group 4, rabbit femoral vein (ED, 2.0-2.2 mm); and group 5, rat femoral vein (ED, 1.0-1.2 mm). In study 2 a graft from the femoral vein was interposed into the carotid artery, with a ratio of the diameter of graft to artery of 1.3:1 in the rats (group 6, n = 12) and 1:1 in the rabbits (group 7, n = 12). In each animal the vessel on one side was repaired using CST and the opposite vessel using the interrupted suture technique. Vessel samples were harvested 1, 2, and 4 weeks after anastomosis. The CST significantly reduced anastomosis time by up to 47% in arteries and 41% in veins. Bleeding time and blood loss were also significantly reduced with CST. Similar results were found in study 2. The total thrombosis rate was 8%, but no significant patency difference was noted between the CST and the interrupted suture technique in any vessel category. We conclude that the CST is a reliable and time-saving procedure in microvascular anastomosis of arteries with diameters greater than 0.7 mm and of veins with diameters greater than 1.0 mm. PMID- 11418921 TI - Effects of major histocompatibility complex matching on graft survival in allogeneic rat limb transplantation. AB - Differences in the major histocompatibility complex (MHC) between recipients and donors present a problem because of immunologic responses in graft rejection. The purpose of this study is to clarify the efficacy of MHC matching against acute graft rejection of allogeneic limb transplants in rats. Right hindlimb transplantations were performed using various MHC-mismatched pairs of inbred rats. The rats were classified into 5 groups according to the differences in subregions of the RT1 (rat MHC) between the recipient and the donor: group 1, RT1 A,B,D barrier (the differences of RT1-A,B,D subregions); group 2, RT1-A barrier; group 3, RT1-B,D barrier; group 4, RT1-B barrier; and group 5, RT1-D barrier. The mean survival time significantly decreased in group 1 and increased in group 4. The results suggest that MHC matching clearly improves survival of transplanted limbs. Specifically, both RT1-A and D matching is the most effective compatibility in prolonging survival time of allogeneic limb transplants in rats. PMID- 11418922 TI - Giant cell tumor of the hand: superior results with curettage, cryosurgery, and cementation. AB - At our institution giant cell tumors arising in all locations are treated with curettage, cryosurgery, and cementation to avoid resection or amputation, increase local tumor control over curettage alone, and avoid the morbidity associated with immobilization. We report the oncologic and functional results of 3 patients with giant cell tumors arising from the tubular bones of the hand who were treated in this manner. At a mean follow-up period of 54 months there were no local recurrences. No patient complained of pain. Digital range of motion and grip strength were within normal limits for all 3 patients. All patients returned to their previous occupational and recreational activities. One instance of minor wound necrosis was successfully treated conservatively. There were no other complications (fractures, infections, neuropraxias, or vascular damage). Curettage, cryosurgery, and cementation performed by experienced surgeons appears to be a safe, effective, and reliable method for treating selected giant cell tumors of the hand. PMID- 11418923 TI - Spontaneous resolution of a phalangeal solitary osteochondroma. AB - We report a solitary osteochondroma in the proximal phalanx of the ring finger in a 3-year-old boy that resolved spontaneously. PMID- 11418924 TI - Comparison between partial and minimal medial epicondylectomy. PMID- 11418926 TI - The injection of nodules of Dupuytren's disease with triamcinalone acetonide. PMID- 11418928 TI - Subchondral bone mineralization patterns in Kienbock's disease. PMID- 11418930 TI - Neuropeptides in insect development and reproduction. PMID- 11418931 TI - Gonadotropins in insects: an overview. AB - Control of gonad development in insects requires juvenile hormone, ecdysteroids, and a peptidic brain gonadotropin(s). Compared to vertebrates, the situation in insects with respect to the molecular structure of gonadotropins is far less uniform. Follicle Stimulating Hormone (FSH) and Luteinizing Hormone (LH) of vertebrates are glycoproteins that are synthezised in the hypothalamus and released from the anterior pituitary. They stimulate gonad development, the production of progesterone or of sex steroids (estrogens, androgens). None of the known insect gonadotropins is a glycoprotein, neither can they be grouped into a single peptide family. In Drosophila, two G-protein coupled receptors, structurally related to the mammalian glycoprotein hormone receptors, have been identified. Nothing is known about their natural ligands. The sex-steroids of insects are likely to be ecdysteroids (20E in females, E in males of some species). Some of the identified gonadotropins speed up vitellogenesis (locust OMP and some -PF/-RFamide peptides) or stimulate ecdysteroid production by the ovaries (locust-OMP and Aedes- OEH) or testis (testis ecdysiotropin of Lymantria). In flies, the only as yet identified gonadotropin is the cAMP generating peptide of Neobellieria. The seeming absence of uniformity in gonadotropins in insects might be due to a multitude of factors that can stimulate ecdysteroid production and/or to the use of different bioassays. Arch. PMID- 11418932 TI - Endocrine mechanisms controlling body-color polymorphism in locusts. AB - The present article reviews recent published and unpublished findings on the hormonal mechanisms for the control of body-color polymorphism in locusts. Emphasis is placed on the dark color-inducing factors and their role in the induction of various types of body coloration observed under different environmental conditions. Implantation of corpora cardiaca (CC) taken from normal nymphs of Locusta migratoria induced dark color in nymphs of an albino strain. Using the albino strain for the bioassay, a neuropeptide, [His7]-corazonin, was identified as a dark color-inducing factor for L. migratoria and Schistocerca gregaria. In the former, depending upon the dose and timing of the injection, this peptide and juvenile hormone developed various body colors looking like those found in nature. The body coloration characteristic of gregarious forms was also induced in isolated albino nymphs and field-collected solitary nymphs. In S. gregaria, on the other hand, the peptide induced black patterns, but the orange or yellow background color observed in gregarious forms was not induced when the peptide was injected into solitary individuals. [His7]-corazonin also induced darkening in other grasshoppers and locusts, but not in katydids. Albino L. migratoria developed dark color when implanted with brains or CC taken from other insects belonging to 10 major insect orders, but not with those from Coleoptera. [His7]-corazonin or a similar compound is widespread among insects and plays a pivotal role in controlling body color in some species and presumably other physiological roles in other species. Arch. PMID- 11418933 TI - Mass spectrometric evidence for the deficiency in the dark-color-inducing hormone,. AB - A factor present in the brain and corpus cardiacum responsible for the induction of dark colour in Locusta migratoria was recently isolated and identified from the corpora cardiaca of normally pigmented locusts. The purification of this factor, designated as [His7]-corazonin was monitored using an albino mutant from a laboratory colony of an Okinawa (Japan) strain. In this study, we provide unequivocal mass spectrometric evidence that the brain and the corpora cardiaca of this albino Locusta mutant are deficient in [His7]-corazonin. Previously, [His7]-corazonin was shown to be responsible for the induction of dark body colour patterns as observed in crowded locusts. Using nanoflow-liquid chromatography-mass spectrometry, we demonstrated that this dark colour-inducing hormone is, however, present in the corpora cardiaca of solitary locusts (Schistocerca gregaria). Arch. PMID- 11418934 TI - Captopril, a specific inhibitor of angiotensin converting enzyme, enhances both trypsin and vitellogenin titers in the grey fleshfly Neobellieria bullata. AB - A strong and constitutive angiotensin converting enzyme- or ACE-like activity was demonstrated in the hemolymph of the adult grey fleshfly Neobellieria bullata. In a competition assay, the N. bullata trypsin modulating oostatic factor (Neb-TMOF) was confirmed to be an in vitro substrate for this circulating Neb-ACE. Oral uptake of captopril, a selective and specific inhibitor of ACE, resulted in a complete phenotypic knockout of circulating ACE activity. When compared with control animals, captopril-fed female flies showed an increase in the liver meal induced trypsin peak in the midgut and elevated levels of protein meal-induced yolk polypeptides in the hemolymph. The latter effect was not due to a slower vitellogenin uptake by the ovaries, because oocyte growth was not affected by the captopril treatment. The apparent synergism between the demonstrated ACE functionality and the previously reported effects of the oostatic peptide Neb TMOF are discussed in the context of our recent finding that Neb-TMOF represents a prime candidate for being the first known in vivo substrate for circulating insect ACE. Arch. PMID- 11418935 TI - Mu-opioid receptors in the ventral tegmental area are targeted to presynaptically and directly modulate mesocortical projection neurons. AB - Mesocorticolimbic projections originating from dopaminergic and GABAergic neurons in the ventral tegmental area (VTA) play a critical role in opiate addiction. Activation of mu-opioid receptors (MOR), which are located mainly within inhibitory neurons in the VTA, results in enhanced dopaminergic transmission in target regions, including the medial prefrontal cortex (mPFC). We combined retrograde tract-tracing and electron microscopic immunocytochemistry to determine if neurons in the VTA that project to the mPFC contain MOR or receive input from MOR-containing terminals. Rats received unilateral injections of the retrograde tracer Fluoro-Gold (FG) into the mPFC. Tissue sections throughout the VTA were then processed for electron microscopic examination of FG and MOR. Immunoperoxidase labeling for FG was present in VTA cell bodies that contained immunogold-silver particles for MOR that often were contacted by profiles exclusively immunoreactive for MOR, including somata and axon terminals. The majority of dually labeled profiles were dendrites that received convergent input from unlabeled axon terminals forming either symmetric or asymmetric type synapses. Within retrogradely labeled cell bodies and proximal dendrites, MOR immunoreactivity was mainly sequestered within the cytoplasm. In contrast, distal retrogradely labeled dendrites contained MOR gold particles located along the plasma membranes. These data suggest that opiates active at MOR in the VTA modulate cortical activity through 1) presynaptic actions on MOR in terminals contacting mesocortical cell bodies, and 2) direct activation of MOR in distal dendrites of projection neurons. PMID- 11418936 TI - Differentially altered mGluR1 and mGluR5 mRNA expression in rat caudate nucleus and nucleus accumbens in the development and expression of behavioral sensitization to repeated amphetamine administration. AB - Altered glutamatergic transmission in the striatum may be implicated in behavioral sensitization to repeated amphetamine (AMPH) administration. Quantitative in situ hybridization histochemistry was performed to define the effects of acute and chronic AMPH exposures on mRNA expression of Group I metabotropic glutamate receptors (mGluRs) in the striatum. Behavioral ratings indicated that the motor activity of rats was significantly higher after the final of five daily AMPH injections (4 mg/kg, i.p.) than that after the first of five daily AMPH, indicative of the development of behavioral sensitization. Moreover, the motor activity of rats treated with five daily AMPH was significantly greater than that of rats treated with five daily saline in response to a 2 mg/kg challenge dose of AMPH 7, 14, 28, and 60 days after the discontinuation of drug treatments, indicative of the persistent expression of behavioral sensitization. Three hours after acute administration of AMPH to naive rats, mGluR1 and mGluR5 mRNA expression in the dorsal (caudatoputamen) and ventral (nucleus accumbens) striatum showed no change as compared to acute saline injection. In rats that developed behavioral sensitization to repeated AMPH, mGluR1 levels in the dorsal and ventral striatum were increased by 53% and 43%, respectively, 3 h after the final AMPH treatment. However, this change did not persist during withdrawal since it was not observed 7, 14, and 28 days after the discontinuation of AMPH treatment. Conversely, mGluR5 levels were markedly reduced 3 h after the final of five daily AMPH treatments in the entire striatum of sensitized rats (34% and 77% of controls in the dorsal and ventral striatum, respectively). The reduction persisted at 7, 14, and 28 days of withdrawal. These results reveal a close linkage between striatal Group I mGluR gene expression and behavioral sensitization to AMPH. This may indicate functional implications of the two subtypes of Group I mGluRs in the regulation of behavioral sensitization to the dopamine stimulant. PMID- 11418937 TI - [(125)I]3beta-(4-ethyl-3-iodophenyl)nortropane-2beta-carboxylic acid methyl ester ([(125)I]EINT): a potent and selective radioligand for the brain serotonin transporter. AB - The binding characteristics of [(125)I]3beta-(4-ethyl-3-iodophenyl)nortropane 2beta-carboxylic acid methyl ester ([(125)I]EINT), a high-affinity selective ligand for the serotonin transporter (5-HTT), and its binding characteristics to rat brain membranes were determined. [(125)I]EINT binding to rat cerebral cortex membranes was saturable and reversible, and its specific binding represented approximately 90% of the total binding. [(125)I]EINT labeled a single site with K(d) = 0.22 +/- 0.03 nM and B(max) = 583 +/- 38 fmol/mg protein. Kinetic analysis revealed a t(1/2) for association and dissociation of 20 and 24 min, respectively. Pharmacological characterization of [(125)I]EINT confirmed its high specificity for the 5-HTT. The pattern of brain region distribution in vivo of intravenously administered [(125)I]EINT indicated greater accumulation of the radioligand in 5-HTT-rich brain regions. However, the signal-to-background ratio was low. Thus, [(125)I]EINT appears to be a useful radioligand for studying the 5 HTT in vitro, but it may not be a good in vivo ligand. PMID- 11418938 TI - Age differences in muscarinic cholinergic receptors assayed with (+)N [(11)C]methyl-3-piperidyl benzilate in the brains of conscious monkeys. AB - Age-related changes in muscarinic cholinergic receptors were evaluated with the novel ligand (+)N-[(11)C]methyl-3-piperidyl benzilate ((+)3-MPB) in the living brains of young (5.9 +/- 1.8 years old) and aged (19.0 +/- 3.3 years old) monkeys (Macaca mulatta) in the conscious state using high-resolution positron emission tomography (PET). For quantitative analysis of receptor binding in vivo, metabolite-corrected arterial plasma radioactivity curves were obtained as an input function into the brain, and kinetic analyses using the three-compartment model and graphical Logan plot analysis were applied. Kinetic analyses of [(11)C](+)3-MPB indicated a regionally specific decrease in the receptor binding in vivo determined as binding potential (BP) = k(3)/k(4) in aged animals compared with young animals. Thus, the frontal and temporal cortices as well as the striatum showed age-related reduction of muscarinic cholinergic receptors in vivo, reflecting the reduced receptor density (B(max)) determined by Scatchard plot analysis in vivo. In the hippocampus, although BP of [(11)C](+)3-MPB indicated no significant age-related changes, it showed an inverse correlation with individual cortisol levels in plasma. When the graphical Logan plot analysis was applied, all regions assayed showed significant age-related decrease of [(11)C](+)3-MPB binding. These results demonstrate the usefulness of kinetic three-compartment model analysis of [(11)C](+)3-MPB with metabolite-corrected arterial plasma input as an indicator for the aging process of the cortical muscarinic cholinergic receptors in vivo as measured by PET. PMID- 11418939 TI - Total regional and global number of synapses in the human brain neocortex. AB - An estimator of the total number of synapses in neocortex of human autopsy brains based on unbiased stereological principles is described. Each randomly chosen cerebral hemisphere was stratified into the four major neocortical regions. Uniform sampling with a varying sampling fraction in each region of neocortex was performed. The total volume of each neocortical region was estimated using point counting according to Cavalieri's principle. The ethanolic phosphotungstic acid staining technique was modified for synapses in human autopsy brains. The numerical density of synapses in each neocortical region studied was estimated using the disector at the electron microscopical level. The total number of neocortical synapses in each region was estimated as the product of the total volume of neocortex and the numerical density of synapses. The influence of the postmortem fixation delay on the number of synapses was investigated in five large mammals (one dog, one cow, and three pigs), the brains of which were kept under conditions similar to those under which human corpses are normally kept. The apparent decrease of 3.9% in the numerical density of synapses in the large mammals following a 2-day fixation delay was not significant. The average total number of synapses in the neocortex of five young male brains was 164 x 10(12) (CV = 0.17). An analysis of the precision of the estimate of the total number of synapses in neocortex indicates that blocks represent both the major source of variation and the largest workload. Using eight blocks per brain the imprecision of the estimate is, however, only 66% of the total variance. PMID- 11418940 TI - Changes en masse. Catholic systems announce three deals including transfer, closure of hospitals. PMID- 11418941 TI - Hardball is OK in S.D. Docs who own competing clinic can lose hospital privileges -state high court. PMID- 11418942 TI - A baby boomer boom. Hospital executives are pinning growth on consumer-oriented age group. PMID- 11418943 TI - Universal to close Texas women's hospital. PMID- 11418944 TI - A sobering challenge. On whim, lawyer buys closed Ill. hospital, now seeks to give it away. PMID- 11418945 TI - MedPAC packs a wallop. Panel opposes bigger Medicare hospital update. PMID- 11418946 TI - Competitors share contracts. PMID- 11418947 TI - The players have changed, but is the game the same? PMID- 11418948 TI - Surveying the landscape. State of Oklahoma using 'score card' to quantify nursing home quality. PMID- 11418949 TI - Mirror images. Two groups of N.Y. hospitals share mirror boards as they look toward merging. PMID- 11418950 TI - Rising rates sink bond sales in 2000. PMID- 11418951 TI - Regulation mania. IRS unleashes long-awaited rules on intermediate sanctions law. PMID- 11418952 TI - Joint effort in Calif. is a first. County health department, federal agencies partner to distribute marijuana to AIDS patients. PMID- 11418953 TI - Less Stark, more clarity. Long-awaited federal regulations may relax strict limits on physician self-referrals. PMID- 11418954 TI - Investigation heats up. Probe of alleged kickbacks at Tenet hospitals escalates. PMID- 11418955 TI - A fresh start. Aetna, now focused solely on healthcare, faces financial, operational challenges. PMID- 11418956 TI - Lone star joining galaxy. Texas launches disclosure of hospital discharge information. PMID- 11418957 TI - Healing through nature. Hospitals cultivate medical, financial interest in on site gardens. PMID- 11418958 TI - The deals are off. Number of hospitals in mergers, acquisitions dropped for second straight year in 2000. PMID- 11418959 TI - 2000 mergers and acquisitions. PMID- 11418960 TI - Trouble in the city. Mergers, Medicare and managed care combine to force closing of 38 urban hospitals. PMID- 11418961 TI - 2000 hospital closures. PMID- 11418962 TI - Web survey. November survey results: a look at medical technology. PMID- 11418963 TI - A do-it-yourself system. Mass. hospital nixes brand-name image archiving products. PMID- 11418964 TI - On a smaller scale. Boutique investment firms ready to feed a hungry market. PMID- 11418965 TI - Not always so settled. HHS following up to make sure providers that paid up are still on the up and up. PMID- 11418966 TI - [2001, a special year for mental health in the world]. PMID- 11418967 TI - [Risk factors for behavioral and emotional problems in childhood: a community study in Uruguay]. AB - OBJECTIVES: To explore some of the demographic risk factors that are associated with the risk of behavioral or emotional problems in children, and to examine the possibility of an association between psychological stress in the parents and psychological problems in their children. METHODS: In this study, the presence of psychological stress in the parents and certain psychosocial characteristics that increase the risk of mental problems in childhood were examined in a sample of Uruguayan children from three communities, two urban and one rural. The study, which was carried out in Ciudad Vieja and Barrio Sur, in the city of Montevideo, and in Colonia de Sacramento, a rural town, comprised 115 children between the ages of 5 and 15. Mothers answered the Child Psychiatric Morbidity Questionnaire (QMPI), an instrument for the detection of behaviors pointing to the presence of emotional problems in children. Both parents also supplied the demographic information requested in the Psychiatric Epidemiology Research Interview Demoralization Scale; they answered the CAGE questionnaire, a screening tool for alcoholism; they responded to the Social Support Network Inventory; and they answered questions about their self-perceived mental health. RESULTS: Fifty-three percent of the children had scores greater than 6 on the QMPI, which indicates the possible presence of behavioral or emotional problems. Fathers' self perception of emotional problems and mothers' feeling of being demoralized showed a statistically significant association with a greater risk of behavioral or emotional problems in their children. CONCLUSIONS: In light of our results, mental health in childhood is a social and public health problem that warrants further epidemiologic study in Uruguay. PMID- 11418968 TI - The Bambui Health and Aging Study (BHAS): prevalence of risk factors and use of preventive health care services. AB - OBJECTIVE: To describe, for the first time in Brazil, the prevalence of risk factors and the use of preventive health care services among adults 60 years and older. METHODS: This population-based study was conducted during 1996-1997 in the state of Minas Gerais, in the town of Bambui, which is located in the municipality of the same name. In 1996 the town had 14,950 inhabitants. For the study, all the town's residents aged > or = 60 years were selected, along with a comparison sample of persons aged 18-59 years. Data were gathered using interviews and physical examinations. RESULTS: Of the 1,742 inhabitants of the town of Bambui aged > or = 60 years, 1,606 (92.2%) participated in the interview and 1,495 (85.8%) were examined (blood pressure, anthropometric measurements, and blood tests). Among the 1,020 younger adults selected for the survey, 909 of them (89.1%) participated in the interview, and 820 of them (80.4%) underwent the physical examination. The prevalence of the following risk factors was higher among older adults than among younger adults: physical inactivity (47.7% vs. 28.0%); blood pressure 140-159 mmHg (systolic) or 90-99 mmHg (diastolic) (25.8% vs. 8.7%); blood pressure > or = 160 mmHg (systolic) or > or = 100 mmHg (diastolic) (19.1% vs. 4.1%); total cholesterol > or = 240 mg/dL (40.6% vs. 9.8%); and blood glucose > or = 126 mg/dL (12.4% vs. 2.2%). The daily consumption of fruits and vegetables was similar among the older adults and the younger ones (51.9% and 51.7%), as was also body mass index > or = 25 kg/m2 (51.6% and 56.2%). Smoking was lower among the older adults (18.7%) than among the younger ones (31.1%). The proportion of participants whose blood pressure had been checked in the preceding year was high (86.8% among the older adults and 61.2% among the younger ones), suggesting that the high prevalence of uncontrolled hypertension was not due to limited access to health care. CONCLUSIONS: For a long time it was believed that the kinds of risk factors that we found in the urban section of the municipality of Bambui were public health problems only in large Brazilian cities. Around 72% of the municipalities in Brazil have < or = 20,000 inhabitants, and these smaller municipalities contain some 19% of the country's entire population. The high prevalence of various risk factors among the older and younger adults in Bambui indicates a great need for health promotion and disease prevention actions. More research is needed to find out whether our results in Bambui reflect the reality of other small towns in Brazil. PMID- 11418969 TI - Improving hepatitis B immunization among high-risk adolescents: a low-cost intervention on the Mexico-United States border. AB - OBJECTIVE: To analyze factors associated with hepatitis B immunization adherence among adolescents attending a sexually transmitted disease (STD) clinic in El Paso, a city in Texas that is on the border between Mexico and the United States of America. METHODS: In this cohort study with 248 adolescents we obtained data on demographics and health beliefs through personal interviews and medical record abstraction. We monitored each of the individuals for 8 months to determine whether the adolescents received a first, second, and third dose of the hepatitis B vaccine. RESULTS: Overall, 32% of the adolescents received the first immunization, 9% the second, and 2% the third. The strongest predictor of receiving either one or two doses was providing the vaccine at the STD clinic as opposed to referring adolescents to a separate vaccination clinic (relative risk (RR) for receiving the first immunization = 7.3; RR for receiving the second immunization = 3.8). Several health beliefs were also associated with receiving vaccinations. CONCLUSIONS: The results of our study indicate that hepatitis B vaccination programs can be improved through such steps as providing vaccinations at a convenient site, educating adolescents about hepatitis B risk factors, and emphasizing the difficulty of treating hepatitis B infection. PMID- 11418970 TI - [Dental care for HIV-positive individuals: fear, prejudice, and professional ethics]. AB - OBJECTIVE: To describe the ethical aspects involved in the dental care provided to patients who are HIV-positive or who have AIDS. METHODS: Literature review (textbooks and MEDLINE and LILACS databases), with an emphasis on the work developed at the School of Dentistry, Federal University of Minas Gerais, Brazil. We examined the social representation of AIDS, the risk of HIV infection during office visits, the refusal to provide care, referral to other professionals without justification, special charges and office visit hours for HIV-positive patients, and the confidentiality of the serological status of the patient. RESULTS: There is still prejudice and ignorance about the risk of HIV and AIDS infection, on the part of dental surgeons and of patients. CONCLUSIONS: An educational project should be undertaken at dental offices and at universities that train new professionals. In addition, the role of national and regional professional associations in providing information concerning ethical aspects involved in the care of HIV/AIDS patients should be reinforced. PMID- 11418971 TI - [Evaluation of exposure to carbon monoxide among kiosk vendors. Valencia, Venezuela]. AB - OBJECTIVE: To study the level of exposure to carbon monoxide (CO), factors contributing to this exposure, and the exposure's potential effects on the health of vendors who have kiosks located on an avenue with heavy vehicular traffic in the city of Valencia, Venezuela. METHODS: We measured the concentration of CO in the air (CO-air) in 16 kiosks in Valencia and also determined the concentration of carboxyhemoglobin (COHb) in the blood of 20 individuals who worked in those kiosks. We also determined the COHb concentrations in a control group made up of 23 professors, students, and workers from Simon Rodriguez University, which is in a town some 120 km from Valencia, in a remote mountainous area without noticeable pollution. RESULTS: In seven of the Valencia kiosks (43.7%) the permissible CO air limits were exceeded. Among the vendors experimental group there was no correlation between the concentrations of CO-air and of COHb. The COHb average for the experimental group (2.9%) was significantly higher than that of the control group (1.6%) and that of the permissible level, according to the analytical method used (< 1.5%). Of a group of symptoms possibly related to CO exposure that were assessed as to whether or not they occurred on a daily basis, all of these symptoms affected a larger percentage of individuals in the experimental group than in the control group. The most common of these symptoms among the experimental group were fatigue and strong headaches. CONCLUSIONS: While the vendors' COHb average does not seem alarming, the CO exposures found could have an adverse impact on these workers' health, especially if they suffered from cardiovascular disorders, or as the result of additive interactions with other environmental pollutants. We recommend carrying out additional studies with more frequent and extended measurements of air quality, to produce information that could be applied in efforts to prevent and control adverse CO health effects. PMID- 11418972 TI - Antigenic and genomic relation between human influenza A (H3N2) viruses circulating in Argentina during 1998 and the H3N2 vaccine component. AB - OBJECTIVE: Due to the lack of correlation from 1994 to 1997 between the A H3N2 component of the influenza vaccine recommended for this period and the circulating viruses in Argentina, we decided to study the antigenic and genomic relationships of the 1998 A H3N2 Argentine circulating strains with the corresponding vaccine component for that year as recommended by the World Health Organization (WHO). METHODS: We selected 18 influenza A H3N2 strains isolated in Argentina during 1998 to carry out an antigenic and genomic study of their hemagglutinin (HA) and neuraminidase (NA) proteins. For the genomic study we added 3 isolates from Uruguay. We compared the Argentine and Uruguayan strains with available reference strains. RESULTS: We found that all 18 strains from Argentina were similar to the A/Sydney/5/97 (H3N2) strain, as opposed to the A/Wuhan/359/95 (H3N2) strain, which was the vaccine component. This result was confirmed by the genomic study. CONCLUSIONS: The approach that we applied in Argentina has improved the quality and quantity of information about influenza in the country. This type of work should be encouraged in other countries in order to help choose the most appropriate vaccine components each year and provide individuals with the best possible protection against influenza. PMID- 11418973 TI - [Poliomyelitis outbreak caused by vaccine-derived virus in Haiti and the Dominican Republic]. AB - In October 2000, the Ministries of Health of the Dominican Republic and Haiti notified two cases of acute flaccid paralysis (AFP) in rural areas, one of them in a 9-month-old female, and the other in a 2-year-old female, respectively. Stool samples that were obtained from these cases, which occurred in July and August 2000, after a 9-year interruption of wild poliovirus circulation in the Western Hemisphere, revealed the presence of type 1 poliovirus. Genetic sequencing, which was later performed at the Centers for Disease Control and Prevention, in Atlanta, Georgia, United States of America, revealed an atypical descendant of the virus used in the manufacture of the oral polio vaccine (OPV), but with 3% genetic divergence with respect to the parent strain. Normally, viral isolates that derive from vaccine components show 99.5% genetic agreement with the parent strain; in wild polioviruses, on the other hand, this agreement is usually less than 82.0%. Thus, the 3% genetic divergence detected in this study suggests that, in areas with low vaccine coverage, the virus used in the vaccine remained in circulation for at least two years, during which it recovered the neurovirulence and communicability of wild poliovirus type 1. This report describes the characteristics and results of the active search for cases of AFP that was sparked by the detection of the two index cases. It also looks at the public health implications of this outbreak for the entire Region of the Americas. PMID- 11418974 TI - Differential pricing of essential drugs. PMID- 11418975 TI - What quality standards are appropriate for rural water supplies? PMID- 11418976 TI - New major histocompatibility complex class IIB genes from nurse shark. PMID- 11418977 TI - Form, function and phylogenetic relationships of mosquito immune peptides. PMID- 11418978 TI - Evolutionary factors in the emergence of the combinatorial germline antibody repertoire. PMID- 11418979 TI - Reactive oxygen species and antimicrobial defenses of invertebrates: a bivalve model. PMID- 11418980 TI - A role for protease inhibitors in immunity of long-lived animals. PMID- 11418981 TI - Suppression of the Drosophila cellular immune response by Ganaspis xanthopoda. PMID- 11418982 TI - Annelid humoral immunity: cell lysis in earthworms. PMID- 11418983 TI - Horseshoe crab hemocyte-derived lectin recognizing specific O-antigens of lipopolysaccharides. PMID- 11418984 TI - A family of C-type lectins in Manduca sexta. PMID- 11418985 TI - Role of tachylectins in host defense of the Japanese horseshoe crab Tachypleus tridentatus. PMID- 11418986 TI - Hemolytic capacity, properties, and activation requirements of cobra venom factor treated chicken serum. PMID- 11418987 TI - Molecular studies of the molluscan response to digenean infection. AB - A classical body of knowledge regarding molluscan immunobiology has developed over the years that indicates an important role for both soluble hemolymph proteins and hemocytes in internal defense. The incorporation of powerful molecular approaches into the toolkit of the molluscan immunobiologist holds considerable promise for allowing a more precise description of the molecules and processes involved. Recent discoveries indicate that the molluscan internal defense system has components that are simultaneously both tantalizingly similar to, and different from, molecules functioning in a defense context in other phyla. Study of molecular components of the internal defense systems of molluscs promises to be a rewarding and surprising field. PMID- 11418988 TI - Insect lectins and innate immunity. PMID- 11418989 TI - Collagenous lectins in tunicates and the proteolytic activation of complement. PMID- 11418990 TI - Immunological properties of heat shock proteins are phylogenetically conserved. PMID- 11418991 TI - Drosophila cellular immune system: Dorothy encodes a UDP glycosyltransferase. PMID- 11418992 TI - Eicosanoids in insect immune signal transduction. PMID- 11418993 TI - Lectins from tunicates: structure-function relationships in innate immunity. PMID- 11418994 TI - Control mechanisms of the prophenoloxidase cascade. PMID- 11418995 TI - Activated prophenoloxidase engaged in the cell clump/cell adhesion of coleopteran insect, Tenebrio molitor larvae. PMID- 11418996 TI - Three kinds of encapsulation-relating factors in coleopteran insect, Tenebrio molitor larvae. PMID- 11418997 TI - A beta-1,3-glucan-binding protein from Manduca sexta. PMID- 11418998 TI - Individual-specific repertoires of immune cells SRCR receptors in the purple sea urchin (S. Purpuratus). PMID- 11418999 TI - Proteolytic activation of prophenoloxidase in an insect Manduca sexta. PMID- 11419000 TI - Hemolymph proteinases in immune responses of Manduca sexta. PMID- 11419001 TI - Cytotoxic reactions associated with insect immunity. PMID- 11419002 TI - Genomic and cDNA sequence of prophenoloxidases from Drosophila melanogaster. PMID- 11419003 TI - The complement system in sea urchins. PMID- 11419004 TI - Paralytic and plasmatocyte spreading peptide from Manduca sexta. PMID- 11419005 TI - Molecular characterization of IL-1-like molecules from lower vertebrates and invertebrates. PMID- 11419006 TI - The detection and isolation of a novel antimicrobial peptide from the echinoderm, Cucumaria frondosa. PMID- 11419007 TI - Isolation and characterization of an IL-1-like protein from Manduca sexta. PMID- 11419008 TI - Clavanins and styelins, alpha-helical antimicrobial peptides from the hemocytes of Styela clava. PMID- 11419009 TI - The discovery and characterization of an interleukin 6 cytokine family antagonist protein from a marine sponge, Callyspongia sp. PMID- 11419010 TI - Phylogeny of vertebrate cytokines. PMID- 11419011 TI - Antimicrobial proteins in crustaceans. PMID- 11419012 TI - Vitiligo: the time has come to offer patients therapy and hope. PMID- 11419013 TI - What's eating you? Millipedes (Diplopoda). PMID- 11419014 TI - Photo quiz. Lichen planus of the nail. PMID- 11419015 TI - Lichen planus, erythema nodosum, and erythema multiforme in a patient with chronic hepatitis C. AB - After identification of the hepatitis C virus (HCV) in 1989, evidence was established supporting its role in the pathogenesis of a number of cutaneous diseases. This evidence ranges from mere epidemiologic associations, such as lichen planus, to molecular biological investigations that have identified the virus in the pathologic tissues of cutaneous vasculitis, vasculitis with mixed cryoglobulinemia, and porphyria cutanea tarda. We describe a 52-year-old man who was diagnosed with chronic hepatitis C, preceding the appearance of lichen planus, erythema nodosum, and erythema multiforme that coincided with the reactivation of viral replication. PMID- 11419016 TI - Deep dermatophytosis: report of 2 cases and review of the literature. AB - Skin infections due to dermatophytes are common and generally associated with a low degree of morbidity in normal hosts. Rare cases have been reported in which the dermatophyte invaded the deep dermis, subcutis, or even internal organs. Two patients, each of whom had clinical and histological findings of a deep or locally invasive dermatophyte infection, are described. This condition typically presents as a nodular eruption that is characterized histologically by suppurative granulomatous inflammation and deposition of organisms in the reticular dermis. Recognition of the potential of dermatophytes for local invasion in susceptible hosts will help ensure proper diagnosis and timely intervention in these cases. PMID- 11419017 TI - Lupus erythematosus profundus: case reports. AB - Lupus erythematosus profundus is a rare manifestation of lupus. We conducted a study to observe the epidemiologic and clinical aspects of lupus erythematosus profundus, its development into other types of lupus or systemic disease, and its response to treatment. Six patients were followed for variable periods. We conclude that lupus erythematosus profundus is a benign form of lupus, distinguishable by systemic manifestations, clinical evolution and resolution, and heterogeneity in clinical manifestations. A skin biopsy is necessary to make an accurate diagnosis. PMID- 11419018 TI - A tattoo and localized lymphadenopathy: a case report. AB - We report the case of a left inguinal lymph node enlargement in a young man. A tattoo was present in close proximity to the lymph node and had preceded its enlargement by several years. The lymph node was removed surgically, and pathologic examination showed it to contain dark pigment material characteristic of a tattoo. The clinical significance of this finding is discussed, and the relationship of tattoo lymphadenopathy to inflammatory skin reactions and to dermatopathic lymphadenopathy is considered. PMID- 11419019 TI - Lasers in dermatology: a review. AB - The revolution in laser technology has had a significant impact on medicine in general and dermatology in particular and has piqued the interest of physicians, the lay public, and the media. Advances in laser therapy have dramatically improved the clinicians' ability to treat cosmetic and noncosmetic skin lesions safely and effectively. The number and variety of skin problems amenable to laser treatment continues to grow. We will provide a review of the major cosmetic and therapeutic applications of laser therapy. PMID- 11419020 TI - Cutaneous anthrax in eastern Turkey. AB - Anthrax, caused by the spore-forming bacterium Bacillus anthracis, is rarely seen in industrial nations but is common in developing countries. Cutaneous anthrax (CA), the most common form of the disease, accounts for 95% of cases and usually develops on exposed sites. This study reviews the clinical and laboratory findings of 21 patients diagnosed with CA during 2 separate epidemics in the Van region of Turkey. All patients had a history of direct contact with infected cattle. The patients, aged 1.5 to 64 years, included 13 females and 8 males. Of the patients, 9 were 15 years or younger. Skin lesions were localized on the hands and fingers in 15 patients, on the face in 3 patients, on the face and finger in 1 patient, on the chest and finger in 1 patient, and on the eyelid in 1 patient. Gram-positive bacillus were noted on Gram stains of material obtained from skin lesions in 2 patients. All but one patient was successfully treated with penicillin; the unresponsive patient was treated with cefuroxime and required plastic reconstructive surgery because of a skin defect on the eyelid. PMID- 11419021 TI - Karate cicatrices. AB - A 46-year-old male presented with linear scars on the dorsal aspects of both hands. The lesions were caused by trauma occurring during the practice of karate. Sports-related scars should be included in the differential diagnosis of scars on the dorsal aspects of the hands. PMID- 11419022 TI - The skeleton in primary hyperparathyroidism: a review focusing on bone remodeling, structure, mass, and fracture. AB - The mechanisms behind the influence of PHPT on the skeleton are closely connected with bone turnover. Throughout life, the skeleton is continuously renewed by bone remodeling, a process which serves the purpose of repairing damaged bone and adapting the skeleton to changes in physical load. In this process, old bone is removed by osteoclastic resorption and new bone is laid down by osteoblastic formation. Bone mass increases with growth in the first decades of life, and around the age of 30 years the peak bone mass is reached. Thereafter, as a result of mechanisms involving bone remodeling, a net bone loss is seen: 1) A reversible bone loss because of increase in the remodeling space, i.e., the amount of bone resorped but not yet reformed during the remodeling cycle. This mechanism leads to decrease in average trabecular thickness and cortical width, and to increase in cortical porosity. 2) An irreversible bone loss caused by negative bone balance, where the amount of bone formed by the osteoblasts is exceeded by the amount of bone resorbed by the osteoclasts at the same remodeling site. Consequently, progressive thinning of trabecular elements, reduced cortical width and increased cortical porosity is seen. 3) Finally, perforation of trabecular plates by deep resorption lacunae leads to complete irreversible removal of structural bone components. Parathyroid hormone, together with vitamin D, are the principal modulators in calcium homeostasis. The main actions of PTH are executed in bone and kidneys. In the kidneys, PTH increases the tubular re-absorption of calcium, thereby tending to increase serum calcium. PTH also induces increased conversion of 25(OH)-D to 1,25(OH)2-D. This last action, enhances intestinal calcium absorption and increased skeletal calcium mobilization, which further adds to the circulating calcium pool. In bone, the "acute" regulatory actions of PTH on serum calcium are probably accompliced via activation of osteocytes and lining cells. A second mechanism of PTH in bone is the regulation of bone remodeling. The action seems to be an increased recruitment from osteoblastic precursor cells and activation of mature osteoclasts. It is supposed that these responses are predominantly mediated indirectly through actions on osteoblast like or nonosteoblast-like stromal cells, as osteoclasts themselves to not have PTH receptors. Bone metabolism and bone mass are studied by biochemical bone markers, bone histomorphometry, and densitometry. As bone markers and bone histomorphometry give information on bone metabolism from different points of view, these methods are preferably combined. Histomorphometry gives detailed information about bone turnover on cellular level, the whole remodeling sequence is described, and the bone balance can be calculated. However, they focus on a small volume, and may, therefore, not be representative for the whole skeleton. On the other hand, studies of bone markers supply general information about turnover in the whole skeleton, but they do not give facts on the bone turnover on the cellular or tissue level and bone balance. Bone densitometry is the principal method in studying bone mass, but valuable information concerning bone structure also comes from histomorphometry. Bone remodeling is considerably increased in PHPT. Studies of bone markers show increase in both resorptive and formative markers, and the increases seem to be of equivalent size. This is in agreement with histomorphometric findings and shows that the coupling between resorption and formation is preserved. By histomorphometry on iliac crest biopsies, trabecular bone remodeling is found increased by 50%, judged by the increase in activation frequency; a measure of how often new remodeling is initiated on the trabecular bone surface. In PHPT, such remodeling activity is repeated about once every year. Reconstruction of the whole remodeling sequence does not show major deviations in lengths of the resorptive and formative periods compared to normal. Furthermore, the amount of bone removed by the osteoclasts during the resorptive phase is matched by the amount of new bone formed by the osteoblasts leading to a bone balance very close to zero. Compared with trabecular bone, the turnover rate in cortical bone is considerably lower, around 10%. Remodeling of the cortical bone takes place at the endocortical, the pericortical, and the Haversian surfaces. Endocortical bone remodeling activities are very similar to trabecular remodeling activities with good correlation between individual parameters. Periosteal remodeling activity is negligible in PHPT, as it is in the normal state. Cortical porosity, which reflects the remodeling activity on the Haversian surface, is increased by 30-65% in PHPT. (ABSTRACT TRUNCATED) PMID- 11419023 TI - Allotransplantation for multiple myeloma. PMID- 11419024 TI - Tumor markers: should we or shouldn't we? PMID- 11419025 TI - ErbB receptors and Ras: upstream, downstream? PMID- 11419026 TI - Tumor markers in breast cancer monitoring should be scheduled according to initial stage and follow-up time: a prospective study on 859 patients. AB - PURPOSE: The purpose of this study was to identify and standardize optimal decision criteria for maximizing the effectiveness of tumor markers in clinical use during the follow-up of patients operated on for breast cancer. MATERIALS AND METHODS: The study was prospectively performed on 859 patients enrolled in 10 institutions. A total of 13,337 determinations of CEA and 14,330 determinations of CA15.3 were available. The median number of samples per patient was 16 for CEA and 17 for CA15.3. The median follow-up was 7 years. Receiver-operating characteristic analysis was used to evaluate the ability of CEA and CA15.3 to discriminate relapses from patients who had no evidence of disease. The diagnostic performances of the two markers were evaluated using decision criteria based on both dichotomic cut-off points and dynamic variations among serial samples. RESULTS: We selected decision levels corresponding to preset levels of 90% and 99% specificity. Patients with CEA and/or CA15.3 levels above the cut-off values were considered positive only if a 1.5-fold increase occurred among the last sample and the mean of the first three samples. According to the different cut-offs used, specificity ranged from 94% to 99% and sensitivity from 48% to 63%. We calculated predictive values using the prevalence expected with reference to the stage of primary tumor and the length of follow-up. Positive predictive values ranged from 1.6% to 93.7%, and negative predictive values from 88.9% to 100%, according to the clinical scenarios and the decision criteria used. The choice of the decision criteria significantly affected positive predictive values within each patient subset. Differences related to time from surgery were still remarkable for every decision criteria (i.e., positive predictive values ranged from 36.6% to 2.8% in node-negative patients according to the year of observation, although the same cut-off point was used). DISCUSSION: The results of the present prospective study show that different decision criteria may provide different diagnostic performances for the same tumor marker and in the same patient. Therefore, we suggest that different decision criteria be settled and used according to the clinical goals. PMID- 11419027 TI - Signal therapy for RAS-induced cancers in combination of AG 879 and PP1, specific inhibitors for ErbB2 and Src family kinases, that block PAK activation. AB - BACKGROUND: Both EGF family ligands and ErbB family receptor kinases act upstream of RAS to induce mitogenesis of normal cells, such as NIH 3T3 fibroblasts. However, oncogenically mutated RAS, such as v-Ha-RAS is constitutively activated and therefore no longer requires these ligands or receptors for its activation. Nevertheless, it up-regulates the expression of these EGF family ligands. To understand the biologic significance of RAS-induced up-regulation of these ligands in both RAS-induced PAK activation and malignant transformation, we have conducted the following studies, based on the previous observations that (1) the N-terminal SH3 domain of PIX selectively binds a Pro-rich domain of 18 amino acids of PAKs, CDC42/Rac-dependent Ser/Thr kinase family, and (2) this specific interaction is essential for both PAK activation and membrane ruffling RESULTS: Using four distinct, cell-permeable, and highly specific inhibitors, namely WR PAK18, which blocks the PAK-PIX interaction; AG 1478, which inhibits ErbB1 kinase activity; and AG 825 or AG 879, which inhibits ErbB2 kinase activity, we demonstrate that (1) the PAK-PIX interaction is essential for v-Ha-RAS-induced malignant transformation; (2) v-Ha-RAS requires not only ErbB1 but also ErbB2, which are activated through two independent autocrine pathways to induce both the PIX/Rac/CDC42-dependent PAK activation and malignant transformation in vitro; and (3) a combination of AG 879 and the Src family kinase-specific inhibitor PP1 suppresses almost completely the growth of RAS-induced sarcomas in nude mice. CONCLUSION: These findings not only change our conventional view on the role of these RAS-inducible ligands and ErbB family receptors (serving as RAS activators) but also suggest a new avenue for the treatment of RAS-associated cancers by a combination of inhibitors specific for ERbB, Src, or PAK family kinases. PMID- 11419028 TI - Taxane-based chemotherapy for patients with carcinoma of unknown primary site. AB - BACKGROUND: The purpose of this study was to determine the long-term follow-up on survival of patients with carcinoma of unknown primary site treated with taxane based chemotherapy in a multicenter community-based setting. PATIENTS AND METHODS: Patients were treated with three sequential phase II trials between 1995 and 1998 as follows: Study I: paclitaxel 200 mg/m2 intravenously (i.v.) Day 1, carboplatin AUC = 6 i.v. Day 1, and oral etoposide 50 mg daily alternating with 100 mg daily days 1-10 every 3 weeks; Study II: docetaxel 75 mg/m2 i.v. Day 1, cisplatin 75 mg/m2 i.v. Day 1, repeated every 3 weeks; Study III: docetaxel 65 mg/m2 i.v. Day 1, carboplatin AUC 6 i.v. Day 1, repeated every 3 weeks. A total of 144 patients (71 on Study I, 26 on Study II, 47 on Study III) were treated (45% had well differentiated carcinoma, 48% had poorly differentiated carcinomas, and 6% poorly differentiated neuroendocrine tumors). The majority of the patients had multiple sites of metastatic disease. RESULTS: Thirty-six percent of all evaluable patients responded to therapy (27% partial and 9% complete responses). The median survival was 10 months with 1-, 2-, 3-, and 4-year survivals of 42%, 22%, 17%, and 17%, respectively. Follow-up ranges from 11 to 50 months. Women survived significantly longer than men. Thirty-one patients remain alive and 14 are progression-free. The primary toxicity was leukopenia with the carboplatin regimens and nausea and vomiting with the cisplatin regimen. A review of the survival of several large previously reported series of patients was compared to results after taxane-based chemotherapy. A compelling argument is made that chemotherapy is superior to best supportive care alone and that taxane-based chemotherapy is superior to other forms of chemotherapy. However, prospective randomized trials will be necessary to definitively demonstrate the superiority of this treatment compared to other therapies for these patients. CONCLUSION: Taxane-based chemotherapy for patients with carcinoma of unknown primary site appears to be clinically beneficial and is associated with long-term survival for a minority of patients at 2-, 3-, and 4-year follow-up. PMID- 11419029 TI - Variations in the use of adjuvant chemotherapy for node-positive colon cancer in the elderly: a population-based study. AB - BACKGROUND: Since 1990, the recommended adjuvant therapy for patients with surgically resected node-positive colon cancer has been 5-fluorouracil (5-FU), usually in combination with leucovorin or levamisole. The purpose of this study is to assess the distribution of adjuvant 5-FU treatment in the elderly. METHODS: The Surveillance, Epidemiology and End Results-Medicare database provides population-based information on cancer patients, representing approximately 14% of the United States population, along with health care utilization data from Medicare claims files. We studied patients with node-positive colon cancer diagnosed between 1992 and 1996 who survived at least 120 days beyond diagnosis (N = 4998). RESULTS: About 50% of elderly patients received 5-FU within 4 months of diagnosis. The proportion of patients treated with 5-FU increased by about 10% from 1992 to 1996. In a multiple logistic regression model, 5-FU treatment was less likely to be given to older patients (compared with those aged 65-69 years, the odds ratio (OR) [95% CI] was 0.82 [0.67-1.00] for ages 70 to 74 years, 0.47 [0.39-0.57] for ages 75 to 79, 0.17 [0.13-0.20] for ages 80 to 84, and 0.04 [0.03 0.05] for ages 85 to 88 years. Non-Hispanic black patients were less likely to be treated than non-Hispanic white patients (OR 0.46 [0.36-0.59]); patients with more than three positive lymph nodes were more likely to be treated than those with three or less, and those with comorbid conditions were less likely to be treated than those without such conditions. CONCLUSIONS: Despite its proven efficacy in reducing colon cancer mortality, 5-FU-based chemotherapy is not widely used among apparently eligible patients over age 65. Efforts are needed to ensure that elderly and non-Hispanic black patients receive appropriate treatment. PMID- 11419030 TI - Clinical applications of a novel sustained-release injectable drug delivery system: DepoFoam technology. AB - The therapeutic effectiveness of drugs is often limited by the inability to sustain therapeutic levels at the target site. Encapsulation of drugs in multivesicular lipid-based particles for sustained release is a novel approach to improving the pharmacokinetics of drug therapy. This paper reviews the preclinical and clinical literature on the applications and potential therapeutic benefits of DepoFoam technology, a novel sustained-release, injectable drug delivery system. DepoFoam formulations of drugs, including anticancer agents (cytarabine, methotrexate, bleomycin, recombinant interferon alfa, 5 fluorouridine-5'-monophosphate, and others), anti-infective agents (dideoxycytidine, 2'-norcyclic guanosine monophosphate, cidofovir, tobramycin, gentamicin, amikacin), analgesics (morphine, bupivacaine), and macromolecules (insulin, interleukin-2), delivered intrathecally, subcutaneously, intraperitoneally, or intralesionally, provide sustained therapeutic levels of drug at the intended target site and reduce systemic exposure and toxicity. Pharmacokinetic studies have demonstrated that DepoFoam particle encapsulation effectively extends the half-life of drugs, thus prolonging the duration of therapeutic drug concentrations in local tissues or in body spaces into which the encapsulated drug is injected. In the case of cell-cycle phase-specific chemotherapeutic agents, such formulations can improve efficacy and therapeutic ratio. DepoFoam is a promising drug delivery system for sustained release of hydrophilic injectable drugs that has a wide range of potential applications in oncology, infectious disease, analgesia, and other therapeutic areas. PMID- 11419031 TI - Advances in chemotherapy for small cell lung cancer: single-agent activity of newer agents. AB - Small cell lung cancer (SCLC) is a common neoplasm with an extremely poor prognosis. Patients with extensive-stage disease have a 5-year survival rate of 1% to 2%. Identification of new active chemotherapy agents is of great importance for the development of more effective treatment for SCLC. Several new drugs that have established a role in the management of non-SCLC in the past decade are also active in SCLC. The mean response rates in untreated versus previously treated patients for these newer drugs are: 26% versus 14% for vinorelbine, 27% versus 14% for gemcitabine, 45% versus 29% for paclitaxel, 22% versus 25% for docetaxel, 39% versus 19% for topotecan, and 50% versus 16% to 24% for irinotecan. A comparison of the response rates of those agents to more established drugs (e.g., cisplatin and etoposide) suggests that the newer drugs are equally or more active in previously treated patients with SCLC. This activity is even more impressive because initial therapy during the past decade has almost always included platinum and/or an epipodophyllotoxin in the regimen. Furthermore, a recent randomized trial showed that the combination of a newer agent (irinotecan) with cisplatin was superior to a standard etoposide and cisplatin regimen in patients with newly diagnosed extensive-stage SCLC. These data support further evaluation of the newer chemotherapeutic drugs, and especially the camptothecins (irinotecan and topotecan) and the taxanes (paclitaxel and docetaxel), in the initial treatment of SCLC. PMID- 11419032 TI - [Screening for Down's syndrome at week 10-14 by measuring fetal nuchal translucency thickness]. AB - Increased fetal muchal translucency (NT) thickness at 10-14 weeks of gestation may indicate underlying fetal chromosomal abnormalities, anatomical anomalies and genetic syndromes. Between January 1997 and May 1998, 1400 women 10-14 weeks pregnant underwent sonographic screening for detection of Down's syndrome (DS). Follow-up was complete in 1208 (86%). Maternal age ranged from 17-44 years (mean 18.0). 87% were found by screening to have a higher risk (1:380) for DS diagnosed at birth. All these fetuses were karyotyped and 8 had chromosomal abnormalities. 2 fetuses with normal NT were diagnosed later as having DS, 1 by the mid gestation triple test and 1 by amniocentesis because of advanced maternal age. Thus sonographic screening identified 8 out of 10 fetuses found to have chromosomal abnormalities at birth. Neonates not karyotyped before birth had no traits at birth that justified chromosomal analysis. Results of this study suggest that NT measurement, combined with maternal age, is an effective 1st trimester screening method for DS in an unselected obstetric population. PMID- 11419033 TI - [Treatment of velopharyngeal insufficiency--a 10 year experience]. AB - Weak and hypernasal speech, along with nasal escape of air, are the main characteristics of velo-pharyngeal incompetence (VPI). We describe 10 years of experience (1989-1998) with surgical treatment of VPI. 51 patients underwent pharyngeal flap elevation. 37 had cleft palate (8 of them submucous), 7 had neuromuscular disorders and another 7 were idiopathic. All underwent evaluation by a speech therapist before and after operation. 25 had further nasal endoscopy and/or videofluroscopy. There was significant speech improvement in volume and clarity in 35 (73%), mild improvement in 13 (27%) and none in 3. The complication rate was 15% and included sleep apnea, wound infection and dehiscence, stridor and bleeding. Complications were correlated with advanced age. VPI should be diagnosed as early as possible to achieve good results and to prevent social problems. PMID- 11419034 TI - [Low prevalence of hepatitis G infection in dialysis patients in Israel]. AB - Prevalence of hepatitis G virus (HGV) infection in the general western population ranges from 0.2-1.5%. In high-risk groups, such as patients with chronic liver disease, hematologic disorders and drug addicts, prevalence is as high as 10% 15%. Dialysis patients have increased rates of HGV infection (6%-50%). We evaluated prevalence of HGV infection among dialysis patients, and the association between HGV infection and hepatitis C virus (HCV) infection. Serum samples were screened for HGV infection by RT-PCR. Screening for HCV infection was performed by an EIA test and confirmed by RIBA and RT-PCR for HCV. Sera were also tested for HBV markers. The study group included all 78 hemodialysis patients and 7 of the 12 peritoneal dialysis patients in our unit during September to November 1997. 4 (5.2%) were HGV-positive but none were peritoneal dialysis patients. 1 of the 12 HCV-positives was also infected with HGV. HGV infection was not associated with duration of dialysis, number of blood transfusions or levels of transaminases. Prevalence of HGV infection among our hemodialysis patients was low (5.2%), but higher than reported for the general population. Prevalence of HGV/HCV infection in hemodialysis patients was low and unrelated to duration of dialysis, number of blood transfusions and levels of transaminases. PMID- 11419035 TI - [Patient satisfaction and hospital services evaluation: comparison of and private patients]. AB - "Sharap" is a private medical service integrated within our public clinic and hospital services. Clients may choose their physician by paying a fee in addition to what their health insurance agency (Kupat Holim) pays for. All other hospital services are supplied to all patients alike. The main purpose of this study was to evaluate the extent to which this declared policy is maintained in practice. During 5 months in 1997, 198 Sharap patients and 198 regular patients were interviewed in the the general surgery, cardiac surgery, ENT, cardiology, newborn and gynecology wards of this hospital. Both groups were similar in regard to cause of hospitalization, gender, age group (10-year age ranges), and length of hospitalization (at least 24 hours). Similar levels of satisfaction with hospitalization in general and with the nursing service and with supportive services were found in both groups. However, Sharap patients were more satisfied with their physicians than regular patients (87% vs 74%, respectively). Similar results were also found using indirect measures of satisfaction. About 86% in both groups reported having achieved the goal of improvement in health. A high proportion of respondents from both groups (82% and 88%, respectively) could not distinguish between Sharap and regular patients in the ward. However, a greater proportion of regular patients (35% vs 21%) wanted more extensive explanations from their physicians regarding their treatment. Sharap patients belonged to higher socio-economic classes than regular patients. Our evaluation indicates that although the Sharap service enables the affluent to choose their preferred physician, resulting in a different doctor-patient relationship, the service does not create a significant feeling of discrimination among hospitalized patients, and does not interfere with the high level of health services available to the public at large. PMID- 11419036 TI - [Fulminant meningococcemia presenting as a gastroenteritis-like syndrome]. AB - Neisseria meningitidis infection (meningococcemia) is very common throughout the world. It usually presents as meningitis or sometimes pharyngitis. A gastroenteritis-like syndrome, with diarrhea, vomiting and abdominal pain, may occur in children but is very rare in adults. Search of the medical literature revealed only 3 such cases, all in young adults. We report an 80-year-old woman who presented with fever, diarrhea and abdominal pain. Meningococcus infection was later suspected, and proved by culture. Although treatment was intensive and included ceftriaxone (Rocephin) and garamycin, she did not respond and died 40 hours after admission. We draw attention to the possibility that what is usually a common symptom can be the first presentation of a serious, often fatal condition. PMID- 11419037 TI - [Left ventricular outflow tract obstruction without left ventricular hypertrophy treated with ACE inhibitors]. AB - We describe a very unusual case of sudden, severe worsening of congestive heart failure which was caused by ACE inhibitors. Diagnosis was made by echocardiogram showing a typical picture of dynamic, left ventricular outflow tract obstruction without left ventricular hypertrophy, which disappeared on discontinuing ACE inhibitors. This phenomenon has already been described as a complication of other drugs such as nitrates, commonly used as provocative tests for latent obstructive cardiomyopathy. To our knowledge ACE inhibitors have not been described as a causative factor. PMID- 11419038 TI - [Multiple organ damage due to cholesterol embolization]. AB - Cholesterol crystal embolization can affect multiple organ systems and mimic other systemic diseases. We describe a 65-year-old woman who had renal failure, diarrhea, transient ischemic attacks and purple toes due to spontaneous cholesterol crystal embolization. PMID- 11419039 TI - [Retinal lesion due to excessive exposure to sunlight]. AB - Retinal damage caused by direct exposure to the sun's rays is well recognized by the ophthalmic community. Although functional ability in solar retinopathy is usually regained within weeks, some suffer long-term visual impairment. Anatomic damage to the retina, even in those who regain full vision, is permanent. We describe 2 cases of solar retinopathy, 1 of which remained with permanent loss of vision. The role of medical education in preventing damage from solar retinal hazards is stressed. PMID- 11419040 TI - [Premature birth and cognitive functioning in adolescence]. AB - Premature infants are considered a high-risk population for developing cognitive dysfunction. Studies have indicated lower cognitive performance among elementary school children born prematurely. We focused on cognitive functioning of such adolescents. This age was chosen because of its critical importance in the development of the individual. 50 adolescents aged 14-16 years born prematurely were compared with 50 born at full-term and matched for gender, age and socioeconomic status. All subjects attended regular schools and did not suffer severe neurological disorders. Cognitive functioning was measured by the Bender Visual Motor Gestalt Test and by 3 subtests from the Wechsler Intelligence Scale for Children (revised WISC-R test). Results revealed that prematurely born adolescents scored lower than those born at term on all measures of cognitive performance. The results are discussed in terms of their developmental meaning and of therapy for the prematurely born. PMID- 11419041 TI - [Evaluation of asthmatic children presenting at the emergency room at Barzilai Hospital]. AB - Bronchial asthma in the pediatric age group has become prevalent recently. Many children who suffer from asthma arrive at the emergency room (ER) with exacerbations which did not respond to medical treatment at home. Between July and December 1997, 136 children 8 months to 14 years of age (61% below 3 years), were studied in our pediatric ER. Investigation included physical examination and pulse oximetry, which were used as guidelines for scoring the children on arrival and post-treatment. Spirometry was done in those who could cooperate. For each patient a detailed questionnaire about medical and sociodemographic factors was filled. Primary pediatricians used mainly beta-agonist and corticosteroid inhalators, while pediatric pulmonologists used mainly inhaled steroids. There was no relationship between severity of attack on arrival at the ER, mode of treatment and speed of recovery in the ER. More children treated by a general pediatrician more were admitted to hospital. Low parental education and paternal smoking were risk factors for recurrent hospital admissions. Our results indicate that parents must be educated to stop smoking, especially those with asthmatic children, and primary pediatricians should be updated with regard to proper treatment and follow-up of asthma. PMID- 11419043 TI - [Relationship between Hebrew and foreign languages]. PMID- 11419042 TI - [Dose-intensive chemotherapy with continuous infusion 5-fluorouracil]. AB - 54 patients with advanced malignancy refractory to chemotherapy were studied to evaluate efficacy and toxicity of continuous infusion of 5-fluorouracil (5FU) given for 3 weeks. We report results of the first 156 courses given in combination with other drugs. 19 (37%) of the 54 responded, including 3 (6%) with complete response. Toxicity was acceptable, with mucositis in 13 (26%) and 3 (6%) with grade II-III toxicity. Results and toxicity profile were compatible with further disease-oriented studies using this dose-intensive program. PMID- 11419044 TI - [Paraneoplastic autoimmune syndromes of the nervous system]. PMID- 11419045 TI - [Autologous bone marrow transplantation in solid tumors in adults: hopes and disappointment]. PMID- 11419046 TI - [CCR5--the HIV co-receptor--now in Israel]. PMID- 11419047 TI - [Et tu Chlamydia! A heart rending story]. PMID- 11419048 TI - [The effect of plasma lipids on kidney function in primary renal diseases]. PMID- 11419049 TI - [Clinical significance of immunomodulation due to antibiotic treatment]. PMID- 11419050 TI - [Piriform muscle syndrome]. PMID- 11419051 TI - [Linguistic purism instead of accepted international medical terms?]. PMID- 11419052 TI - [Harold Hirschsprung (1830-1916)]. PMID- 11419053 TI - [Antidepressive effect of pyridoxine (vitamin B6) in neuroleptic-treated schizophrenic patients with co-morbid minor depression--preliminary open-label trial]. AB - BACKGROUND: Minor depression is reported in 20-60% of schizophrenic patients during various stages of their disorders; impairing patients' compliance, response to treatment and worsening their overall prognosis. Various anti depressive treatments have been proposed for such cases but response rates are usually poor. Pyridoxine (Vitamin B6) in essential for the proper metabolism of various neurotransmitters that are considered relevant to the pathophysiology of depression and/or schizophrenia and it has been reported beneficial in ameliorating depressive symptoms as part of major depression, premenstrual syndrome or 'Chinese restaurant syndrome'. We hypothesized that addition of pyridoxine to on-going neuroleptic treatment could improve minor depression in schizophrenic patients. METHOD: Nine schizophrenic patients with co-morbid minor depression participated in this study. All participants had a stable unchanged clinical state (changes in Brief Psychiatric Rating Scale (BPRS). Scale for the Assessment of Positive Symptoms (SAPS), and Scale for the Assessment of Negative symptoms (SANS) scores < 5%) and all were maintained on unchanged doses of anti psychotic drugs for at least 4 consecutive weeks prior to initiation of the study. Participants received, open-label, pyridoxine 150 mg/day in addition to their anti-psychotic treatment for 4 consecutive weeks. Mental status was evaluated before, during, and at the end of 4 weeks of pyridoxine administration using the BPRS, SAPS, SANS and HAM-D. RESULTS: Two of the nine patients (22%), characterized by higher initial HAM-D and SANS scores, and by older age and longer duration of illness, experienced marked improvements in depressive symptoms (23% and 28% decrease in HAM-D scores) following 4 weeks of pyridoxine administration. In one of these two, the improvement in depressive symptoms was accompanied by a parallel decrease in SANS Scores. CONCLUSION: A subgroup of schizophrenic patients with comorbid minor depression may benefit from pyridoxine addition to their on-going anti-psychotic treatment. PMID- 11419054 TI - [Choice of infusion site--differences in the references of staff (nurses and residents) and patients]. AB - Insertion of an intravenous (i.v.) line is a common and routine procedure in hospitalized patients. The literature recommends not to insert an i.v. line in a limb fold, but other than that there are no guidelines regarding optimal locations for the i.v. line. In this study we attempted to elucidate whether there are other possible preferences and guidelines for choosing the location of an i.v. line. We studied four groups: patients at the time of hospitalization, hospitalized patients, nurses and surgical interns on a surgical ward. We found that 91.8% of the patients request to take part in choosing the location of their i.v. line. Moreover, we found that most patients have a hand preference for the i.v., whereas, the ward staff has little location preference. Despite these findings, we discovered that, in practice, most patients were not asked to participate in the decision making process regarding their i.v. line location. Analysis of the results showed that there is a need to include the patient's preference in choosing the location for the i.v. line, whenever possible. If the patient doesn't have a preference, the i.v. line should be inserted in the non dominant hand or the hand in the bed which is more accessible to the medical & nursing staff. PMID- 11419055 TI - [Primary gastrointestinal stromal tumors]. AB - Stromal tumors of the GI tract are rare. In the retrospective and prospective study we investigated the relationship between tumor symptomatology, tumor grade and prognostic factors. During the period May 1993-September 1999, 11 female and 13 male patients with a mean age of 62 (range-29-81) years were operated for primary gastrointestinal stomal tumors (GIST) in our department. Observed signs and symptoms were: GI bleeding (65%), abdominal pain (45%), abdominal mass (15%) and weakness (5%). In 4 patients tumor was an incidental finding during investigation or operation for another tumor. Tumor location (in decreasing order) was: stomach (15), small bowel (SB, 6), esophagus (1), duodenum (1) and colon (1). Preoperative biopsy or FNA were diagnostic in less than 50% of the cases. Operative procedures included wedge resection (8 patients), resection of segment of bowel (10) and extended resection (6), of diaphragm, SB, colon, bladder, kidney and liver. The mean tumor size was 7.8 (range-0.9-22) cm. Four tumors were graded as benign, 8 of indeterminate malignant potential and 12 malignant. CONCLUSION: The main presentation of GIST is acute GI bleeding. Endoscopy is most effective for studying proximal tumors, and CT should be used to identify distal GI tract tumors. Tumor size or malignancy were not necessarily predictive of GI bleeding. When invasive to adjacent organs is present, wide excision should be contemplated as long-term survival can be achieved. PMID- 11419056 TI - [Traumatic brain injury--the National Trauma Registry]. AB - BACKGROUND: Traumatic Brain Injury (TBI) has been established as a category in reporting systems. Uniform data systems case definition has been suggested for hospital discharge data surveillance systems cases based on ICD-9-CM diagnostic codes. These include fractures and specific mention of intracranial injuries such as contusion, laceration, hemorrhage, and concussion. Inspection of data from the Israel National Trauma Registry suggested that two diagnostic groups of very different severity and outcome were being unjustifiably combined. AIM: To evaluate the validity of categorizing TBI into two discrete groups, using the presence of specific mention of intracranial injury and/or loss of consciousness for more than one hour as the definition of definite TBI. Possible TBI includes skull fractures with no mention of intracranial injury and/or concussion with no loss of consciousness. METHODS: The study population includes all traumatic injuries admitted to hospital, dying in the ER or transferred to other hospitals and recorded in the 1998 Trauma Registry in all 6 level I trauma centers in Israel and two level II centers. RESULTS: The significant difference in severity between groups supports the validity of sub-dividing the TBI classification into definite and possible subcategories. As a result, we obtain two different severity groups without measuring specific severity scores which are limited in the reporting system. CONCLUSION: The groups were significantly different in severity, hospital resource use, immediate outcome, demographic and injury circumstances. PMID- 11419057 TI - [The effect on family members of treating home-hospitalized patients]. AB - BACKGROUND: In recent years, chronic patients being treated at home are being joined by increasing numbers of acute and sub-acute patients. Although there has been a considerable expansion of formal services for home hospitalization, nevertheless a significant part of the burden of caring for these patients falls on their families. Many studies have examined the effect on the family of treating chronic patients at home, but very little is known about the effects on the family caregiver of treating an acute or sub-acute patient at home. GOALS OF THE STUDY: a. To determine the main areas of care involving family members within the framework of home hospitalization. b. To assess the caregivers' sense of burden. c. To identify the causes of the feelings of burden. d. To examine the caregivers' specific difficulties. e. To identify the areas in which they are particularly interested in additional guidance. METHODOLOGY: The study population included 179 primary caregivers of patients registered in four home hospitalization units. The data were collected during face-to-face interviews with the primary caregivers, in the patient's homes. FINDINGS: Half (49%) of the caregivers were the patients' spouses, and the rest were other family members, 67% were women. The average age was 56, and about a third were over 65. On average, they were involved in six areas of care. Fifty three percent reported that the burden they had to contend with was too heavy, 28% said it was not so heavy, and 19% said it was not heavy at all. A multi-variate analysis revealed that the feeling that the burden was too heavy had a statistically significant correlation with caring for another family member at home, being involved in 3+ areas of treatments, being married, 9+ years of education, the patients suffering from frequent pain or unable to reach the bathroom independently. Most (75%) of the caregivers reported three or more areas of burden which include the time devoted to care giving, and the effect of the treatment on the patient's health, mental and physical condition. Although 77% of them had received instruction on how to treat the patients, 29% cited at least one area in which they would have liked to receive more instruction, noting in particular the need to learn how to deal with changes in the patient's mental status. CONCLUSIONS: Family members caring for home-hospitalized patients require more instruction and advice from the formal agencies during the daily care of the patient, particularly in specific subjects. There is a need to develop additional instruments for advice and instruction, in a broad range of areas, in order to assist the caregivers in their complex role. There is a need to continue to examine specific areas in which the family members require additional counselling and instruction, in order to better plan these services, and help improve the quality of care for home hospitalized patients. PMID- 11419058 TI - [Intravenous immunoglobulins treatment of patients with Graves' ophthalmopathy]. AB - Graves' ophthalmopathy is an autoimmune disease manifested as exophthalmus, lid lag and diplopia. As in the accompanying autoimmune thyroid disease, there is an autoimmune homonal and cellular attack on the orbita, mainly the retro-orbital tissues. Steroids are the cornerstone of therapy. We reviewed the evidence for a similar therapeutic effect of i.v., immunoglobulins (IVIGs) and their better side affect profile as compared to steroids. We also described an impressive therapeutic success with IVIG given to a patient with resistant ophthalmopathy. The clinical picture of Graves' ophthalmopathy is attributed to a pathologic hyper--activation of orbital fibroblasts, deposition of collagen and glycosaminoglycans in the extra-cellular matrix and eventually fibrosis. These are mediated by leucoregulin, IL-1, IFN-gamma, and TGF-beta--all secreted by lymphocytes and mast cells in the retorbital space. Another mode of cell activation is by binding of autoantibodies (presumably thyroid stimulating Ab's) to an antigenic determinant on the surface of fibroblasts. I.v. immunoglobulins, known today to be active in a variety of autoimmune processes, exert their effect on autoantibodies, complement, phagocytic cells etc. IVIGs also inhibit orbital lymphocytes and fibroblasts through inhibition of IL-1 or/and TGF-beta. PMID- 11419059 TI - [Surgery in patients over 100 years of age--5-year experience (1995-2000)]. AB - The age distribution of a given national population is of utmost importance when dealing with public health and analyzing the use of various national health facilities. This is based on the totally different use of health sources by different age groups. Despite continuous prolongation of life expectancy and the related aging of the western hospitalized population the sub-group of patients over 100 year old of age is unusual demographically and accounts for only a very small portion of the total number of patients hospitalized in surgical wards. During the 5 year experience between 1995 and 2000 seventeen such patients were admitted to our surgical departments. Eight of those underwent 11 operations with zero peri-operative mortality and no significant complications. These 11 operations in this elderly population are the basis of our report. PMID- 11419060 TI - [Local necrosis of finger following stab with needle used to pump terbutaline sulfate (Bricalin)]. AB - Terbutaline sulfate (Bricalin) is a widely used medication for asthma. It works mostly, but not only, on beta-adrenergic receptors. In this case study we describe a 50-year-old nurse referred to the emergency department after she was stabbed by needle that was used to draw bricalin for inhalation. She arrived at the emergency room with an inflamed necrotic area at the point of the stab. Later, she developed acute infection that was treated with antibiotics. Following the acute phase there was still a necrotic area, that required surgical debridement. It seems that this is the result of the vasoconstrictive influence of terbutaline sulfate via alpha adrenergic receptors. In the medical literature there is scarce data regarding this side effect. PMID- 11419062 TI - [Harefuah peer review]. PMID- 11419061 TI - [The influence of the presence of students on the consultation--attitudes of tutors in family medicine clinics]. AB - BACKGROUND: The consultation is the pivot of clinical teaching in ambulatory care. It is therefore essential that students observe the consultation. The students' presence itself influences the consultation and also requires the patients' consent. Moreover the introduction in Israel of the "Patients' Rights Act" in 1996 has made us more acutely aware of the place of the patient in teaching especially with regard to the consent to be part of the teaching process. AIM: This study was undertaken in order to investigate how tutors in family medicine perceive changes in the consultation caused by the presence of students. METHODS: An anonymous physician questionnaire was distributed on the first day of the 6th year clinical clerkship in family medicine. The questions pertained to perceived influence on length and content of the consultation. In addition physician and patient background information was gathered; and the physicians were asked to estimate the patients' willingness to be part of the teaching process. RESULTS: 46 tutors in family medicine participated, 70% of whom were female. Sixty four percent of the doctors thought that the student's presence had an influence on the consultation. Ninety one percent thought that it increased consultation length, especially of the physical part (93%). More than half thought that the student's presence might interfere with asking intimate questions. The majority held the opinion that the patient's gender and socioeconomic background were inconsequential. Ninety two percent of physicians estimated that 5% or less of the patients would refuse the presence of a student. IN CONCLUSION: Tutors in family medicine think that the presence of a student affects the consultation. Those involved in and responsible for teaching should take this into account. Further research of these changes with objective measurements is needed. PMID- 11419063 TI - [Cellular phones, a test for the need of credible scientific research]. PMID- 11419064 TI - [Adrenal mass in patients with previous history malignancy: a diagnostic and patient managing challenge]. PMID- 11419065 TI - [Febrile seizures]. PMID- 11419066 TI - [DNA microsatellites and various human diseases]. PMID- 11419067 TI - [The use of imaging modalities in patient with thyroid nodule: ultrasonography and radionuclide scintigraphy with new tracers]. PMID- 11419068 TI - [The role of the medical team in emergency units in case of traumatic loss of permanent anterior teeth]. PMID- 11419069 TI - [Mifepristone--a new antiprogesterone in obstetrics and gynecology]. PMID- 11419070 TI - [Hemorheological aspects of hyperlipidemia]. PMID- 11419071 TI - [Everybody wants to live longer--but who wants to get older? Reflections in the books of Dr. David Cohen: chapters on gerontology]. PMID- 11419072 TI - [Mental health services in Israel--reforms and directions]. PMID- 11419073 TI - [The mouse on the hill]. PMID- 11419074 TI - [Dr. Heinrich Hoffman and "Struwwelpeter"]. PMID- 11419075 TI - [Past editors of Harefuah: Dr. J. L. Rokach 1890-1950]. PMID- 11419076 TI - [The Germans and holocaust]. PMID- 11419077 TI - [Role of polymorphic variants of the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene in the pathogenesis of cardiovascular diseases]. PMID- 11419078 TI - [Left ventricular diastolic dysfunction in chronic heart failure: pathogenesis, diagnosis, treatment]. PMID- 11419079 TI - [Bacterial endocarditis of the tricuspid valve]. PMID- 11419080 TI - [Effectiveness of systemic immunoglobulins in the treatment of the systemic variants of juvenile rheumatoid arthritis]. AB - Preparations of intravenous immunoglobulin (sandoglobulin, pentaglobin, intraglobin F, octagam) were given daily or each other day in a course dose 0.3-1 g/kg to 43 patients aged 4 to 15 years. Eight of them had allergosepsis, 22- allergoseptic variant of juvenile rheumatoid arthritis (JRA) and 13--systemic JRA and generalized joint syndrome. The treatment induced remission in patients with allergosepsis, prednisolone was discontinued. JRA patients coped with fever, eruption, polyserositis, infection symptoms. Leukocyte count normalized. Preparations of intravenous immunoglobulin in low doses can be therapy of choice in patients with rheumatic-like diseases. In JRA patients it can be effectively used against fever, eruption, polyserositis, carditis, leukocytosis, intercurrent infection before or in the course of immunosuppressive therapy. PMID- 11419081 TI - [Comparative assessment of the left chest EKG leads V7-V9 and bipolar Slapak and Partilla leads in the diagnosis of posterior and posteriobasal myocardial infarction]. AB - The study included 100 patients with acute inferoposterior myocardial infarction consequently admitted to hospital. Both the standard ECG in 12 leads as well as additional V7-V9 and Slapak & Partilla leads were recorded in each patient. Two dimensional echo at rest was performed to evaluate regional wall motion. The criterion of dividing patients into any MI subgroup was the presence of abnormal Q wave in any system leads. Posterior + inferior MI was found in 88%, limited posterobasal--in 2% and diaphragmal (inferior)--in 10% of patients. Good correlation of Q-wave in II, aVF and S1-S4, V7-V9 and S1-S4 was found. The more abnormal Q waves were revealed in lead systems the more was necrosis size as assessed by echo. We conclude, that additional lead systems are of value in revealing necrosis size and its location in patients with damaged posterior wall. PMID- 11419082 TI - [Clinical significance of stress-protein content in peripheral blood lymphocytes of patients with myocardial infarction]. AB - We studied the role of inducible stress-protein with molecular mass 70 kD (heat shock protein--HSP) in myocardial protection in ischemia. Content of HSP was measured in 25 patients with Q myocardial infarction (QMI) and 5 healthy controls. Western-blot was made on QMI day 1, 2, 7, 14 and 21, and once in the controls. Three types of HSP synthesis reaction manifested on QMI day 1. Type 1- HSP was absent in lymphocyte cytoplasm at 37 degrees C and after heat shock (HS); type 2--HSP accumulated after HS and was absent at 37 degrees C; type 3--HSP was present at 37 degrees C and after HS. Healthy subjects exhibited HSP after HS and the HSP synthesis was enhanced by HS. Type 1 was associated with favourable short term prognosis of MI. Vast myocardial damage was accompanied with lowering synthesis with its activation at 37 degrees C by MI day 2. High synthesis of HSP on MI day 2-14 indicated moderate heart damage. Thus, HSP reactivity can be used as a factor of MI prognosis reflecting mobilization of intracellular defense proteins. PMID- 11419083 TI - [Correlation of somatotropic hormone with post-infarction myocardial remodelling and clinical prognosis in patients with myocardial infarction]. AB - To evaluate pathogenetic link of ischemic heart dysfunction with the level of endogenic STH in myocardial infarction (MI) patients at different MI stages and to specify prognostic value of STH for MI outcomes, we examined 78 males on MI day 21-26, month 3-6 and a year later. 20 healthy males served control and 17 patients with ischemic heart diseases without MI according to coronaroventriculography (CVG) were a comparison group. STH was measured in all the patients with radioimmunoassay, CVG was made according to M. Judkins. We found that in patients with high level of endogenic STH (> 1 nm/l) within the first postmyocardial infarction month development of reparative and adaptive fibrosis continues 3 to 6 months and is more favorable. If STH physiological level maintains, myocardial contractility remains satisfactory for a long period. In high STH levels, patients with myocardial ischemia more frequently exhibit ventricular arrhythmia. Low STH blood levels lead to left ventricular dilatation, deterioration of cardiac contractility, stenotic coronary atherosclerosis, intracardiac hemodynamics and IHD clinical course. Lethal outcomes occur more frequently. PMID- 11419084 TI - [Prostaglandin E1: results and perspectives in clinical practice]. PMID- 11419085 TI - [Use of magnetosorbents in the immunodiagnosis and therapy of systemic lupus erythematosus and rheumatoid arthritis]. AB - The authors present new technology of producing immobilized granulated antigenic magnetosorbents (MS) using collagen type II, IgG, denaturated DNA and cardiolipin which were employed in enzyme immunoassay for detection of specific antibodies in the blood of patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). Due to high capacity, MS demonstrated effectiveness in early diagnosis, prognosis of RA and SLE as well as in control over the treatment effects. As shown in in vitro experiments, MS have high selective capacity and atraumaticity for blood corpuscles. MS can be sterilized, regenerated and reused which is a great advantage. PMID- 11419086 TI - [Exercise training of patients in a health resort after surgery for rheumatic heart disease]. AB - We studied the effect of cardiovascular system function on the response to terrain cure in the course of rehabilitation of 88 patients after surgical correction of valvular defects in conditions of low-mountain health resort. We have found that the response to the above exercise depends on functional condition of the cardiovascular system (the degree of correction of hemodynamic disorders related to valvular disease, the degree of myocardial hypertrophy and dilation of the chambers, left ventricular contractility). Terrain cure exercise promotes improvement in regulation of cardiovascular system and mobilization of its reserves. PMID- 11419087 TI - [Antibodies to beta2-glycoprotein-1 in patients with systemic lupus erythematosus]. AB - Antibodies to beta 2-glycoprotein I (beta 2-GPI) and antibodies to cardiolipin (aCL) were determined by enzyme immunoassay in 58 patients with verified SLE. Elevated level of antibodies to beta 2-GPI was associated with development of antiphospholipid syndrome (APS) and aCL hyperproduction. It is evident that synthesis of antibodies to beta 2-GPI is an additional marker of APS in SLE. PMID- 11419088 TI - [Prognostic criteria in the course of duodenal ulcer]. AB - Examination and long-term follow-up covered 95 patients with duodenal ulcer (DU). Mean age of the patients was 32 +/- 1.67 years. The patients were divided into two groups: with rare DU recurrences (n = 36) and with frequent recurrences (n = 59). Diagnostic criteria for development of rarely recurrent DU were the following: blood group A(II), exacerbations in winter, weak symptoms at DU onset, weak abdominal pains, solitary surface ulcers. For development of frequently recurrent DU, diagnostic criteria include O(I) blood group, spring exacerbations, marked symptoms of the exacerbation, severe abdominal pains, multiple deep ulcers. PMID- 11419089 TI - [Tolerance and safety of anti-Helicobacter drugs in the treatment of ulcer disease]. AB - 132 patients (mean age 46.9 +/- 1.4 years) with exacerbation of gastroduodenal ulcer (GDU) positive by Helicobacter pylori (HP) were examined clinically and using laboratory tests. All the patients received treatment with anti-HP drugs. Side effects were registered in 81(61.4%) patients. Negative trend in colon biocenosis occurred in all the patients. The treatment was poorly tolerated by women, aged patients and those with ulcer in the stomach. Biocenosis aggravation is indication for administration of eubiotics. PMID- 11419090 TI - [Effectiveness of anti-ulcer agents and the extent of gastric mucosa contamination with Helicobacter pylori]. AB - Effectiveness of H. pylori eradication was studied in 56 ulcer patients (46 males, 10 females). It was compared to the degree of gastric mucosa contamination with H. pylori. A correlation was found between H. pylori eradication, time of ulcer healing and severity of the infection. Patients with high contamination demonstrated the least frequency of eradication and the longest time of healing. It is suggested to perform eradication treatment of H. pylori adjusting its duration to the degree of the bacterial contamination. PMID- 11419091 TI - [Circadian rhythm of external respiration in patients with bronchial asthma treated with Melaxen]. AB - Melaxen treatment of bronchial asthma (BA) patients produced positive effects on biorhythms of external respiration. Quantitative characteristics of volume and speed rhythms change insignificantly. The basic quantitative effect consists in internal and external synchronization and elimination of ultradian rhythms indicating stabilization in the system of control and coordination of time structure of external respiration system. PMID- 11419092 TI - [Isosorbide-5-mononitrate (mono mac) in the prevention of bleeding from esophageal varicose veins in patients with liver cirrhosis]. AB - Varicose esophageal veins bleeding is one of severe complications of hepatic cirrhosis. This complication is effectively managed with combination of beta blockers with nitrates. Of the latter, optimal is isosorbite-5-mononitrate (mono mac). Its minimal risk of development of nitrate tolerance and pharmacokinetics does not depend on hepatic or renal function. A single dose of mono mac varies from 10 to 50 mg. The individual dose is selected by a fall in systolic blood pressure at rest (by 15-20 mm Hg) above 100 mm Hg. In appearance of serious headache nitrates are followed by beta-blockers monotherapy. It is inferred that isosorbite-5-mononitrate (mono mac) is effective in prophylaxis of hemorrhage from varicose veins of the esophagus both in beta-blockers and as monotherapy when beta-blockers are contraindicated. PMID- 11419093 TI - [Attitude of patients after myocardial infarction to long-term therapy with beta blockers]. AB - 107 patients were followed up for 12 months after myocardial infarction. Their own and their doctors' attitude to long-term intake of beta-blockers (BB) was asked in the end of each month using questioning or questionnaires. Only 22% of the patients continued BB treatment by the end of the year. The primary reason of this situation was rejection of the patients. Hospital doctors often do not prescribe long-term BB because of overestimation of the side effects and contraindications. Doctors of outpatient clinics do not like the necessity to keep the patients under continuous care. Some physicians have poor knowledge of the role of BB in secondary prevention of myocardial infarction. PMID- 11419094 TI - [Clinical variants of Wegener's granulomatosis]. PMID- 11419096 TI - [Effective surgery]. PMID- 11419097 TI - [Neuroradiological methods in ophthalmology]. PMID- 11419098 TI - [Medical decisions]. PMID- 11419099 TI - ["Runny nose and earache"--a torment for the child, a burden for the community]. PMID- 11419100 TI - [Nasal congestion and earache--upper respiratory tract infections in 4-year-old children]. AB - BACKGROUND: Few population-based studies in Norway have addressed upper respiratory infections in children. MATERIAL AND METHODS: A questionnaire concerning health, diseases and living condition was administered to parents of all four-year-old children in Vestfold county (n = 1912). Participation rate was 79%. RESULTS: Close to one out of three of the children suffered from recurrent common cold (four episodes or more) during the last year. One third experienced otitis at least once, and one out of twenty experienced four episodes of otitis or more. Day-care centre attendance was the only significant risk factor for recurrent common cold (odds ratio 1.50; 95% CI 1.18-1.92) or otitis (odds ratio 1.42; 95% CI 1.12-1.77). The odds ratios were essentially unchanged after logistic regression analysis, when mother's education, housing, other children in the family, passive smoking, and history of breastfeeding were included in the analysis. Children with otitis or recurrent common cold consulted a doctor almost ten times as often as other children. INTERPRETATION: Day-care centre attendance has significant influence on the occurrence of upper respiratory infections in four-year-olds, but less influence than reported for younger children. Other environmental differences seem to be of little importance and do not represent any potential for prophylaxis. PMID- 11419101 TI - [Hereditary thyroxin-binding globulin deficiency--changed thyroid function tests]. AB - BACKGROUND: Variation in concentrations of carrier proteins of hormones may influence the effect of the hormones and may cause confusion in the interpretation of laboratory results. MATERIAL AND METHOD: A Caucasian family with a hereditary thyroxin-binding globulin (TBG) deficiency was investigated. 22 persons in two generations had blood tests for TBG, thyrotropin (TSH), three iodothyronin (T3), thyroglobulin (TG), thyroxin and for free thyroxin (FT4) by two different commercial tests, Delfia and IMx Abbott (IMx). Relevant health information was collected of all persons. RESULTS: Six males had very low T4 values, non-detectable TBG, increased FT4 values on the Delfia test and within normal range on the IMx test. Six females had lower borderline T4 and TBG. All persons were clinical euthyroid. INTERPRETATION: The condition is considered to represent X-chromosome linked inheritance with hemizygote affected males and heterozygote female carriers with intermediate values for T4 and TBG. Commercial test kits for FT4 may present considerably different results in conditions with TBG deficiency. When a high level of measured FT4 combined with normal TSH is found; TBG deficiency should be considered. PMID- 11419102 TI - [Metastasis to the parotid gland]. AB - BACKGROUND: The skin areas that primarily drain into the parotid lymph nodes include large parts of the face, cheek, anterior surface of the auricle, temporal region, scalp and forehead. Malignant tumours, mainly squamous cell carcinomas and malignant melanomas, may therefore metastasis to the parotid gland. It may, however, sometimes be difficult to decide whether the parotid tumour is a primary or secondary tumour. MATERIAL AND METHODS: Twenty-six patients treated for metastatic malignant disease to the parotid gland were evaluated retrospectively. The mean age of the 22 males and four female patients was 70 years. RESULTS: In nine patients (35%), the primary tumour was ignored and wrongly interpreted as a tumour originating in the parotid gland. The primary tumour was localised in the temple in six patients, in the cheek in six, in the anterior surface of the outer ear in five, in the eyelids in four, in the forehead in one, in the scalp in one, and in the nasal vestibulum or nasal cavity in three patients. 19 patients (73%) had squamous cell carcinomas and seven (27%) malignant melanomas. The treatment was surgery supplied by radiotherapy in 19 patients. Four (21%) of the patients with squamous cell carcinoma and four (57%) of those with malignant melanoma died from the disease. INTERPRETATION: When a tumour arises in the parotid gland, the physician should critically evaluate whether this represents a secondary tumour. Careful examination of the scalp and face for suspect skin lesions and scars from extirpation of previous skin cancer can help detect the primary tumour. PMID- 11419103 TI - [Airborne dust particles in indoor environment and allergy]. AB - BACKGROUND: The overall aim of this study was to investigate how airborne house dust particles may contribute to an allergic immune response, and thereby also to asthma and allergic diseases. MATERIAL AND METHODS: Using transmission electron microscopy, we quantified and characterized airborne house dust particles, with regard to elemental and size distribution. Furthermore, an immunogold labelling technique was used to study whether some common allergens were present on the surface of airborne house dust particles. Finally, a mouse model was used to study the adjuvant activity of airborne house dust on the IgE antibody response. RESULTS: A vast majority of the airborne particles samples from homes in Oslo were found to be less than 2.5 microns in diameter, thus they are liable to penetrate deep into the respiratory tree. This PM2.5 fraction contained, in addition to sulphur aerosols and silicates, many soot particles, most of them being less than 1 micron in diameter. These soot particles were found to carry allergens on their surface. We also found that diesel exhaust particles, which is probably a main soot component of airborne house dust, absorbed several wellknown allergens in vitro. Furthermore, the airborne house dust particles were found to elicit a local lymph node response, and to have an adjuvant activity on the production of IgE antibodies to ovalbumin as a model allergen. INTERPRETATION: These results show that indoor suspended particulate matter contains a lot of potential allergen carriers, i.e. soot particles (carbon aggregates), most of them being less that 1 micron in diameter and thereby able to transport allergens deep into the airways. In addition, our results indicate that suspended particulate matter may have an adjuvant effect on the production of IgE to common environmental allergens, and also may provoke a local inflammatory response. PMID- 11419104 TI - [Choice of neuroradiological methods in ophthalmology--the eye and the lacrimal apparatus]. AB - With improved imaging methods, the possibility of demonstrating pathological changes in the visual pathways has increased substantially. However, optimal evaluation of visual disorders requires familiarity with the anatomy and pathology of the visual pathways, and with the many advances in neuroimaging. The purpose of this article is to provide clinicians with a practical approach for selecting the most appropriate imaging modalities. Choice of technique is discussed with reference to anatomical regions rather than a complete list of diseases. Ophthalmoscopy reveals many intraocular abnormalities. Imaging studies help in cases where opaque media preclude a view of the fundus. In addition, imaging studies assist in confirming the extraocular extent of the lesion. The advantages and disadvantages of x-ray, computed tomography and magnetic resonance imaging are discussed and illustrated by examples. PMID- 11419105 TI - [Choice of neuroradiological methods in ophthalmology--the optic nerve, muscles of the eye and orbit]. AB - Optimal imaging of the orbital structures presents a challenge. Images of this region may be disturbed by ocular movements, and orbital fat may prevent visualisation of the optic nerve and retrobulbar lesions. In this article, we intend to provide practical guidelines for the evaluation of orbital abnormalities, on the basis of our own experiences and a study of selected literature. Computed tomography (CT) and magnetic resonance imaging (MRI) are both useful for the detection and characterisation of orbital abnormality. However, for diagnostic work-up of the optic nerves, contrast-enhanced, fat suppressed MRI best demonstrates the pattern and severity of optic nerve abnormality and allows assessment of the intracranial part of the optic nerves and associated intracranial abnormality. In the case of bony involvement, spiral CT imaging is the modality of choice. It is quick, inexpensive and allows multiplanar reconstruction. Not uncommonly, CT as well as MRI may be needed to characterise an orbital lesion. PMID- 11419106 TI - [Neuroradiological imaging of the 3rd, 4th and 6th cranial nerves]. AB - The ocular motor cranial nerves (III, IV, VI) control the eye movements in a near association with the higher cortical areas. Clinically, the most common presentation of abnormal ocular motility is double vision. Identifying the cause of ocular nerve palsy can be difficult, and a large percentage of such cases still remains undiagnosed, even in a new era of neuroradiological techniques. Close co-operation between the clinician and the radiologist is necessary for the selection of the best imaging methods for the specific clinical problem, in order to set the aetiological and topological diagnosis. This article provides a practical review of advances in neuroimaging of the ocular motor nerves. PMID- 11419107 TI - [Suspected large fetus in the last period of pregnancy--a difficult problem]. AB - BACKGROUND: In pregnancy surveillance a large symphysis-to-fundus measure raises several questions concerning delivery. MATERIAL AND METHODS: We review various problems with large for gestation age foetuses, also called macrosomic foetuses. We have performed literature searches mainly through PuBMed, which includes the Medline database. The clinical problem is discussed from the primary care provider's point of view and from those of the patient and the obstetrician. RESULTS: Macrosomia is defined as foetal weight above the 90th percentile, birth weight above 4000 g or 4500 g, or birth weight over + 2 SD of the mean birthweight by age. The diagnosis is difficult, even with various sonographic procedures. Abdominal circumference alone appears to have the same diagnostic value as the use of a combination of biparietal diameter, femur length and abdominal circumference. INTERPRETATION: Based on the literature, labour should not be induced or caesarean section performed in non-diabetic pregnancies unless the estimated foetal weight is above 5000 g. A great number of caesarean sections would have to be performed to avoid a single case of plexus brachialis paresis due to difficult shoulder delivery. The best policy is to await spontaneous birth or to induce birth after the completion of 42 weeks. In pregnancies complicated by diabetes mellitus, there are reasons for selective induction of labour if macrosomia is suspected, and for caesarean section if the calculated birth weight is above 4000 g. As the problem of difficult shoulder delivery cannot be completely avoided, each department should have a strategy to handle such a situation. Various procedures for managing the difficult shoulder delivery are described. PMID- 11419109 TI - [Eduard Boeckmann--scientist, inventor and benefactor]. AB - Eduard Boeckmann (1849-1927) left a lasting legacy in Norway and in America. His scientific career started when as a medical student he won the Skjelderup gold medal for a study of the tonsils. In Bergen he presented his thesis, an experimental study dealing with the cause of keratitis, which affected many patients with leprosy. Later he published vigorously from his medical practice. In St. Paul, Minnesota, he constructed a steam autoclave based on principles that became fundamental in later autoclave technique. His autoclave was put into industrial production. In his own laboratory he worked with improvement and safe sterilization of catgut, and catgut was produced in St. Paul for 59 years, till 1960. He donated income from the catgut production to a fund for a medical library, today the Boeckmann Library of the United Hospital in St. Paul. Boeckmann had a high reputation as a doctor. His American patients were first of all Norwegian settlers in the north-western states. Both in Bergen and in St. Paul he was highly active in the professional associations. PMID- 11419108 TI - [Comparison of preventive effect of "new" and "old" antihypertensive agents]. AB - BACKGROUND: Until recently no morbidity-mortality study had examined the effects of newer drugs like angiotensin-converting enzyme inhibitors, calcium-antagonists and alpha-blockers compared to "old" but well-proven thiazide diuretics and beta blockers in the treatment of essential hypertension. MATERIAL AND METHODS: The prospective and randomized clinical trials CAPPP, STOP-2, NORDIL, INSIGHT and one arm of ALLHAT, with a total of approximately 58,000 middle-aged or elderly hypertensive patients have been assessed. RESULTS: The primary outcome, composite cardiovascular (CV) death, cerebral stroke and myocardial infarction, in one study with heart failure, or composite fatal coronary heart disease and myocardial infarction, was equal in all trials. INTERPRETATION: According to current evidence, prevention of cardiovascular disease in hypertension is the same irrespective of the class of drug. PMID- 11419110 TI - [NSAID preparations for external application--do they have any effect and should they be used?]. PMID- 11419111 TI - [Risk, uncertainty and ignorance in medicine]. AB - Exploration of healthy patients' risk factors for disease has become a major medical activity. The rationale behind primary prevention through exploration and therapeutic risk reduction is not separated from the theoretical assumption that every form of uncertainty can be expressed as risk. Distinguishing "risk" (as quantitative probabilities in a known sample space), "strict uncertainty" (when the sample space is known, but probabilities of events cannot be quantified) and "ignorance" (when the sample space is not fully known), a typical clinical situation (primary risk of coronary disease) is analysed. It is shown how strict uncertainty and sometimes ignorance can be present, in which case the orthodox decision theoretical rationale for treatment breaks down. For use in such cases, a different ideal model of rationality is proposed, focusing on the patient's considered reasons. This model has profound implications for the current understanding of medical professionalism as well as for the design of clinical guidelines. PMID- 11419112 TI - [Knowledge must be combined with values for decision making in uncertainty]. AB - On the basis of a clinical presentation of a woman with atrial fibrillation and diabetes mellitus, this article examines how medical evidence can be combined with patient preferences in the analysis of clinical choice. Such choices tend to be analysed and discussed without due consideration for patient or public values. It is argued that such an analysis is incomplete until the patient's preferences are incorporated into the model. This is also a way of implementing patient centred medicine. We discuss how decision theory can be used to clarify clinical choices, sometimes with surprising results for the physician. How to handle situations when the patient's preferences are in conflict with the physician's judgement, is also discussed. It is concluded that good clinical practice must be based on evidence, but it should also take into account the patient's values in choices with uncertain outcomes. PMID- 11419113 TI - [Shift and escalation of professional liability]. AB - Doctors are at risk of having to take blame even if they have acted according to good medical practice. One reason for this is to be found in two mechanisms, shift of liability and escalation of liability--irrational mechanisms by which doctors are affected after a medical mishap. A mishap is difficult to accept and those who suffer because of it want to place responsibility somewhere. A shift of liability occurs when responsibility is displaced from the patient or the system to the doctor. Escalation takes place when a mishap is considered much worse after the fact than in the risk evaluation ex ante. The author points at the problems these mechanisms cause for patients, doctors, and society and provides some suggestions as to how doctors may handle such situations in a fruitful manner. PMID- 11419114 TI - [Understanding the function of brain--a historical perspective]. PMID- 11419115 TI - [A museum, like this, all cities should have]. PMID- 11419116 TI - [Incontinence following emergency surgery and ileoanal anastomosis in ulcerative colitis]. PMID- 11419117 TI - [Do general practitioners (and others) know too little about chronic pain?]. PMID- 11419118 TI - The Microcystis cyanobacteria bloom in the Swan River--February 2000. AB - In February 2000 the Swan-Canning estuary in Western Australia experienced a record bloom of the toxic cyanobacteria Microcystis aeruginosa. At its height, concentrations of M. aeruginosa reached integrated water column cell counts of 15,000/ml and formed bright green scums in sheltered bays, where counts of 130 million cells/ml were recorded. Due to public health concerns parts of the river were closed from 10 to 22 February 2000. Two unseasonably large summer rain events in early and late January 2000 created conditions for the bloom. Freshwater runoff, estimated at 270 GL, was enough to fill the Swan-Canning estuary five times over and brought with it high levels of nutrients, mainly nitrogen (> 2.0 mg/L TN) and phosphorus (> 0.15 mg/L TP). A number of methods to reduce bloom accumulations were tried, including an attempt to increase the salinity of the surface water above the critical 10 ppt level for Microcystis; using a bentonite clay and poly-aluminium chloride mixture to flocculate and sink the algae; and sucking up scums using oil spill equipment. Over 900 tonnes of M. aeruginosa were removed and safely disposed using sewage treatment facilities. The bloom collapsed when the freshwater flush subsided and seawater intrusion from the Indian Ocean re-established itself, raising the salinities above the tolerance of Microcystis. PMID- 11419119 TI - River loggers--a new tool to monitor riverine suspended particle fluxes. AB - The Australian Institute of Marine Science (AIMS) has developed an in situ digital turbidity logger (AIMS River Logger) to record time series of fine suspended sediment concentrations in North Queensland rivers. The loggers use dual LED transmissometers (15 and 85 mm pathlength) to measure in situ turbidity associated with the range of suspended sediment concentrations (0-5 g L-1) encountered in regional rivers. A system of wiper brushes clean the optical surfaces prior to instrument readings. Overlying water depth and temperature are concurrently recorded. Internal batteries provide sufficient power for unattended deployments of 6+ months with readings taken at 30 minute intervals. Records of suspended sediment concentrations in the lower Tully River over three wet seasons (1996, 1997, 2000) are shown in relation to concurrent river discharge. Peaks in suspended sediment concentration coincide with discharge events. During each event, peak sediment concentrations typically occur during the rising stage of the hydrograph, then decline rapidly prior to significant falls in water level. Derived suspended loads during discharge events are generally proportional to the size of the event, though higher responses are recorded during the initial flood event of each wet season. PMID- 11419120 TI - Community and connectivity: summary of a community based monitoring program set up to assess the movement of nutrients and sediments into the Great Barrier Reef during high flow events. AB - The Great Barrier Reef (GBR) system encompasses the largest system of corals and related life forms anywhere in the world. The health of this extensive system, particularly the inshore area, is dependent on the relationship between the GBR and adjacent coastal catchments. The major impact of agricultural practices on the GBR is the degradation of water quality in receiving (rivers) waters, caused by increased inputs of nutrients, suspended sediments and other pollutants. For the past three years, the Great Barrier Reef Marine Park Authority (GBRMPA) has been involved with the co-ordination of a river-monitoring program, specifically targeting the sampling of rivers during flood events. Representative sites were set up along two North Queensland rivers, the Russell-Mulgrave and Barron Rivers. This monitoring program is run in conjunction with the Queensland Department of Natural Resources' Waterwatch program. The program involves intensive sampling of first flush, extreme flow and post flood conditions over the two rivers. Extreme flow conditions are sampled over a limited time span (48 hours) with trained volunteers at 4-hour intervals. Concentrations measured in the flood events are dependent on land use characteristic, and extent of flow. Concentrations of dissolved and particulate nutrients are higher if the extreme flow event is part of the first flush cycle. Concentrations of DIN and DIP measured before, during and after a major flood event suggest that there is a large storage of inorganic material within the Barron and Russell-Mulgrave agricultural subcatchments that move over a period of days, and perhaps weeks. This program created a forum in which GBRMPA liased with the Barron and Russell-Mulgrave community about the connectivity existing between the river and the Great Barrier Reef lagoon. PMID- 11419121 TI - The use of artificial oxygenation to reduce nutrient availability in the Canning River, Western Australia. AB - Artificial oxygenation has been used for two summer periods to improve the water quality of the Canning River in Perth, Western Australia. The project is part of the Swan Canning Cleanup Program, which aims to reduce the frequency and severity of nuisance and toxic algal blooms in the Swan-Canning estuary. The trials have proved that oxygenation has increased the dissolved oxygen concentrations in the water column, particularly in the bottom waters where dissolved oxygen concentrations are frequently below a critical level of three milligrams per litre. Oxygenation has had a positive impact on nutrient concentrations in the water column and nitrogen cycling processes. Reductions in nutrient concentrations were highlighted by drops in ammonium and total phosphorus concentrations of 97% and 64% following the recommencement of oxygenation after a plant shutdown. Results of a microbiological study combined with the data analysis indicate that the number of nitrifying microbes have increased due to oxygenation. However, comparisons between oxygenated and control areas were inconclusive about the ability of the oxygenation plant to reduce total nitrogen and phosphorus levels. This could be explained by factors such as spatial variability, water flow during the trials and measurement limitations in the monitoring program. Future work will concentrate on assessing the impact of the oxygenation plant on nutrient concentrations. PMID- 11419122 TI - Classifying urban rivers. AB - Classification systems have been developed over the last century as a tool to aid managers in the preservation, conservation, enhancement and management of rivers. The classification systems developed to date have been designed to differentiate between relatively unimpacted, mainly rural rivers. Urban rivers typically show poor water quality and biological diversity, and so most current classification systems tend to group urban rivers into a single "poor" category. In this paper we describe a hierarchical framework for recording information about urban rivers that allows a more sensitive description of these rivers enabling subdivision into several classes according to the purpose of the classification. The different levels in the hierarchy, the types of attributes that are to be recorded at each level, and the relational database structure for storing the data are described. The 100-500 m river stretch level in the hierarchy relates to the engineered modification of urban rivers and is the key to their classification. An example classification at this scale illustrates a link between engineering modification, bank and bed materials and the number and diversity of physical habitats present. This classification underlines the importance of adopting a hierarchy of nested spatial scales for data collection, classification and interpretation since it illustrates a clear link between characteristics at the stretch scale and at the finer habitat scale. The classification also illustrates the varied nature of urban rivers and the fact that even quite heavily engineered stretches can contain a diversity of habitat types. PMID- 11419123 TI - Managing fish, flood plains and food security in the Lower Mekong Basin. AB - The "Lower Mekong Basin" in this paper refers to the part of the Mekong River Basin which is shared by Cambodia, Laos, Thailand and Viet Nam, all members of the Mekong River Commission, consisting of approx. 2,400 km of mainstream river, numerous tributaries and huge flood plains. Few river basins produce as much fish as the Mekong River Basin, and the fishery in the Lower Mekong Basin is among the biggest and most productive inland fisheries in the world. The flood plains of the Lower Mekong produce some four times as much fish per square kilometre as the North Sea, which is among the most productive marine areas in the world. It is quite clear that the fisheries in the Mekong Basin are very important for the population in respect to their food security and income. Its importance in nutrition is highest in the rural areas, where there are few other low cost sources of protein, and even in highland areas fish is of crucial importance in the diet. Most fish species in the Mekong Basin are migratory, and the economically most important ones are certainly so. However, with economic development gaining speed, the impact on migratory patterns and the competition for the water resources are becoming stronger. The water resources offer a large number of opportunities, and a lot of economic activities need access to the water resources for their development. However, what is seen in one sector as an opportunity may be considered as a threat in another, and a careful balance is necessary in order not to lose opportunities in important sectors. The fate of a large number of river basins in the world is frightening. Most have been left biologically near dead, with some of the big rivers reduced for a time, or forever, to be used as waste water canals for the new industries, and others almost dried out from excessive water extraction before they reach the sea. PMID- 11419124 TI - Australia versus the world: do we face special opportunities and challenges in restoring Australian streams? AB - Australians do face special opportunities and challenges in attempting to restore or rehabilitate their streams, when compared with Western Europe and North America. Some of these differences arise from the physical and ecological character of our streams, particularly in terms of flow variability, stream power, sediment, vegetation interactions, and nutrient levels. We also have modest resources available per-unit-length of stream, and no massive freshwater fishing industry. These, and many other differences, combine to produce an environment in which it can be slower and more difficult to rehabilitate streams, and we have less confidence in the results. International case studies are tremendously useful but in some cases we need to be circumspect in applying the results directly to Australian streams. PMID- 11419125 TI - Meaendering: nature, culture and rivers. AB - Alongside science we need a parallel capability of exploring meaning so that intangible, uncountable aspects of our relationships with rivers, especially those understood by local communities, can be expressed. Common Ground has offered the concept of Local Distinctiveness and gives examples from England, including recent projects which they have initiated. PMID- 11419126 TI - Forecasting rehabilitation outcomes for degraded New Zealand pastoral streams. AB - To understand the timescales and magnitude of responses that can be expected following catchment and riparian rehabilitation, we forecast changes to selected stream ecosystem attributes following tree planting in a pastoral catchment. All planting scenarios were predicted to lead to decreases in daily maximum water temperature after 15-20 years to levels that would be suitable for sensitive invertebrate species. Cooling and reheating were rapid so that most benefits to water temperature along the mainstem were forecast to accrue from shading all of the stream channel network. All planting scenarios were predicted to increase sediment yields over the status quo over the 25-year timeframe examined, with maximal sediment yield occurring about 15 years after planting due to expected erosion of the streambanks under the developing forest shade. Sediment yield was greatest for full catchment planting over 25 years, although sediment yield would be lowest with this scenario over longer timescales. A macroinvertebrate biotic index was predicted to increase by 25% over 15 years if whole catchment afforestation were implemented, compared to 9% if only the 4th order mainstem were planted with riparian trees. The use of ecological forecasting to predict likely outcomes for a range of scenarios should prove useful for prioritising rehabilitation actions. PMID- 11419127 TI - Informing river management policies and programs with science. AB - Conventional wisdom has it that we already have enough science to address the problems causing degradation of our environment, including rivers. This is not true. However it is the case that we could be using existing knowledge better, and that we could be doing more to learn the lessons from the huge sums being spent on river restoration and management. Informing good policy and practical on ground management with R&D outputs has proven to be is difficult, but essential. This paper reviews some of the history of water and river management in Australia and how perceptions of rivers have evolved. It discusses the challenge of enhancing the linkages between science, policy and practice in river management. It outlines the knowledge exchange, R&D and capacity building strategies of the National Rivers Consortium--a new initiative whose founding partners are LWRRDC, the MDBC, CSIRO Land and Water and the Western Australian Waters and Rivers Commission. This strategic collaboration between policy makers, river managers and scientists brings together organisations with responsibility and expertise to improve the health and management of Australian rivers. The National Rivers Consortium is making a major investment in knowledge exchange and capacity building, based on direct personal contact and learning by doing. The Consortium is establishing a program of training activities targeting river managers and policy makers, based on the best available science and high quality information products. It will support river managers as they plan and implement river restoration and protection projects. The paper concludes with a discussion of the key knowledge gaps that remain impediments to the better management of Australia's unique and diverse river landscapes. PMID- 11419128 TI - Towards integrated catchment management, Whaingaroa, New Zealand. AB - The paper examines progress towards integrated catchment management and sustainable agriculture at Whaingaroa (Raglan), New Zealand. Application of the Canadian "Atlantic Coastal Action Program" model (ACAP) has been only partially successful within New Zealand's bicultural setting. Even before the introduction of the ACAP process there existed strong motivation and leadership by various sectors of the community. A merging of resource management planning and implementation processes of the larger community and that of the Maori community has not occurred. Research carried out by Crown Research Institutes has clearly shown the actions required to make pastoral farming more sustainable. There are difficulties in the transference to, and uptake of, these techniques by farmers. An examination of the socio-economic context is required. There has been a requirement on local government bodies to tighten their focus as part of recent reform. This has occurred concurrently with a widening of vision towards integrated and sustainable forms of management. This (as well as a clear belief in empowerment of local communities) has lead to Council reliance on voluntary labour. There is a need to account for the dynamic interaction between social and political history and the geological and biophysical history of the area. As part of a re-examination of sustainable development, New Zealand needs to reconcile the earning of the bulk of its foreign income from primary production, with the accelerating ecological deficit that it creates. A sustainability strategy is required linking consumer demand, property rights and responsibilities. PMID- 11419129 TI - Catchment management and the Great Barrier Reef. AB - Pollution of coastal regions of the Great Barrier Reef is dominated by runoff from the adjacent catchment. Catchment land-use is dominated by beef grazing and cropping, largely sugarcane cultivation, with relatively minor urban development. Runoff of sediment, nutrients and pesticides is increasing and for nitrogen is now four times the natural amount discharged 150 years ago. Significant effects and potential threats are now evident on inshore reefs, seagrasses and marine animals. There is no effective legislation or processes in place to manage agricultural pollution. The Great Barrier Reef Marine Park Act does not provide effective jurisdiction on the catchment. Queensland legislation relies on voluntary codes and there is no assessment of the effectiveness of the codes. Integrated catchment management strategies, also voluntary, provide some positive outcomes but are of limited success. Pollutant loads are predicted to continue to increase and it is unlikely that current management regimes will prevent this. New mechanisms to prevent continued degradation of inshore ecosystems of the Great Barrier Reef World Heritage Area are urgently needed. PMID- 11419130 TI - Institutional barriers and opportunities: processes and arrangements for natural resource management in Australia. AB - While "institutional failure" is commonly cited as a core barrier to sustainable resource management, operational prescriptions or criteria for institutional design are rare. Institutions are crucial, and may be barriers or opportunities. This article offers some principles and design features for "adaptive" institutions, capable of balancing the core principles of persistence, purposefulness, information-richness, inclusiveness and flexibility. It then draws on emerging findings from recent Australian research to identify some major institutional barriers and opportunities for improving capacities in sustainable resource management. PMID- 11419131 TI - Establishing an Integrated Catchment Management (ICM) program in East Java, Indonesia. AB - The Brantas is one of Indonesia's most important catchments. It is the "rice bowl" of Java and nationally important for its industrial activity. Surabaya, Indonesia's second largest city, is located at the mouth of the Brantas River which is pivotal to the city's water supply. The challenges associated with the institutional framework for natural resource management in East Java parallels that of many states and provinces around the globe. It is multi-layered and complex. Integrated Catchment Management (ICM) may be defined as "the co ordinated and sustainable management of land, water, soil vegetation, fauna and other natural resources on a water catchment basis". Over a period of six months, an ICM Strategy was researched and facilitated for the Brantas River Catchment in East Java via a short term advisor attachment. The aim of the Strategy is to improve coordination, co-operation, communication and consistency of government and community efforts towards sustaining the catchment's environmental, economic and social values. The attachment was part of the Pollution Control Implementation (PCI) Project funded by AusAid and the Indonesian Government. The ICM Strategy developed was broad based and addressed the priority natural resource management issues facing the Brantas Catchment. It was co-ordinated by BAPEDALDA, the Provincial Environmental Protection Agency, and developed by all agencies involved in natural resource management in the catchment. Various Universities and Non Government Organisations (NGOs) were also involved in the ICM process which developed the Strategy. At the conclusion of the attachment, a draft ICM Strategy and a proposed institutional framework had been developed. A working group of key agencies was also established to further enhance local "ownership", finalise timescales and implementation responsibilities within the Strategy and bring the institutional arrangements into being through a Governor's Decree. PMID- 11419132 TI - Indigenous property rights and river management. AB - The presence of indigenous property rights and interests arising from the survival of native title in Australia presents unique issues in the management of rivers and riverine lands. Existing common law and statutory tidal and non-tidal rights are a complex overlay of public and private property rights which are themselves undergoing significant change through the commodification of many natural resources by Commonwealth and State governments, such as marine species stock and non-tidal water. The melding of indigenous values and management practices with existing management regimes for rivers and riverine lands offers considerable potential for both sustainability of resource utilisation, and respect and recognition of native title with resultant predicted benefits in the vexed area of compensation. PMID- 11419133 TI - From here to accountability: securing healthy rivers. AB - While specifying "responsibilities" for river planning and management implies that outcomes are desired, defining "accountabilities" demands that results are achieved. This paper examines innovative ways of converting aspirations to results, with particular reference to the changes initiated in New South Wales in response to the establishment of the Healthy Rivers Commission of New South Wales and its first round of inquiries. PMID- 11419134 TI - From river to ridge: a blueprint for sustainable management in the Georges River catchment. AB - Research on sustainable transport and water projects, conducted in North America during 1999, identified four key considerations common to a number of projects. These four considerations are the preconditions necessary to achieve sustainability in a given location. These preconditions (BAPP) are boundary definition, administrative alignment, protection of non-urban lands, and the participation of all interested parties. This paper considers the perceived attitudes of various catchment stakeholders and focuses on planners and planning. The results of questionnaire surveys based on BAPP concepts suggest that local government is perceived as having the highest priority for protection of riparian lands and waterways (as individual Councils and Regional Organisations of Councils). Thus local government is the preferred arena for coordination, responsibility and jurisdiction of waterways and riparian land. The opportunity exists to examine the differing roles of local and state government. The paper concludes by outlining the implications of this research for land use planning and institutional structures in sustainable river management. PMID- 11419135 TI - Establishing river basin organisations inVietnam: Red River, Dong Nai River and Lower Mekong Delta. AB - River basin management is receiving considerable attention at present. Part of the debate, now occurring worldwide, concerns the nature of the organisations that are required to manage river basins successfully, and whether special purpose river basin organisations (RBOs) are always necessary and in what circumstance they are likely to (i) add to the management of the water resources and (ii) be successful. The development of river basin management requires a number of important elements to be developed to a point where the river basin can be managed successfully. These include the relevant laws, the public and non government institutions, the technical capabilities of the people, the understanding and motivation of people, and the technical capacity and systems, including information. A river basin organisation (or RBO) is taken to mean a special-purpose organisation charged with some part of the management of the water resources of a particular river basin. Generally speaking, such organisations are responsible for various functions related to the supply, distribution, protection and allocation of water, and their boundaries follow the watershed of the river in question. However, the same functions can be carried out by various organisations, which are not configured on the geographical boundaries of a river basin. This paper outlines recent work on river basin organisation in Vietnam, and makes some comparisons with the situation in Australia. PMID- 11419136 TI - Managing waterway health in the Goulburn Broken Catchment, Victoria, Australia. AB - Historically within most catchments, resource management programs have been planned and implemented in isolation of one another. This was once the case in the Goulburn Broken Catchment, a major catchment of the Murray Darling Basin, Australia. Although only 2% of the Murray Darling Basin's land area, the catchment generates 11% of the basin's water resources. Learning from the past, a cooperative and collaborative approach to natural resource programs has developed. This approach is the envy of many other catchment communities and agencies. Through a combination of "Partnership Programs", "Operational Initiatives" and community involvement, significant programs have been implemented within the catchment, which will benefit not only the local community but communities further afield. The outcomes of the waterway health program highlight the benefits provided through the establishment of cooperative and partnership resource improvement programs. These programs were founded on the ability of the community to recognise the need for integration, base management decisions on best available science and an ability to work together. Their effective delivery has been provided through the resources provided, to the local community, by the Natural Heritage Trust with matching and State and local allocations. While programs have shown success, challenges still face the community. These challenges include verification and implementation of environmental flows, storage of the catchment's vital water resources, and maintaining community involvement and participation in on-going works programs. The Goulburn Broken Catchment community, with the support of Federal, State and Local Governments, is looking at opportunities for continued improvements in waterway health. PMID- 11419137 TI - Sustaining our rivers in crisis: setting the international agenda for action. AB - The history of streams and rivers is as much a social and technological history as it is a scientific one. Rivers are the lifeblood of nations and the control of their waters has been fundamental to the building of human civilisations. The control or regulation of rivers embodied the advancement of institutions, administration and co-ordination; it was a manifestation of military and economic power. Yet the history of human development is also characterised by the degradation of the basic resource--polluted water, increased flooding, and the loss of biological diversity. Many early civilisations collapsed in the face of environmental degradation, manifest by flood, drought, famine and plague. The Industrial Revolution upon which modern societies are founded was based upon a short-term vision that has left rivers in crisis, marked by a legacy of pollution, slums, a loss of confidence in civic life, and a loss of ownership of places and spaces--once seen to be at the heart of civilized society. Within this global or international context of water management, this paper examines the impacts and future of rivers and water within the United Kingdom, establishing some principles for such management in other settings. PMID- 11419138 TI - Challenges of the Blackwood Basin, Western Australia. AB - Growing concern about the declining state of the catchment and river led to the formation of the Blackwood Basin Group in 1992. Funded primarily by the Natural Heritage Trust and using the river as the focus, the group aims to provide leadership and support to achieve sustainable management of natural resources in the catchment. Through an Integrated Catchment Management approach, the Blackwood Basin Group has managed a range of projects to improve the community's understanding and management of the Blackwood River and its catchment. A number of research, education, demonstration and on-ground action activities relating to river management have been undertaken in partnership with community and local, state and federal government organisations. Activities include demonstrations and evaluations of riparian restoration, funding riparian restoration activities, protection of high conservation value remnant vegetation, a flood risk study, zone action planning and monitoring the condition of the river and its tributaries. PMID- 11419139 TI - Achievements of the Grand River Conservation Authority, Ontario, Canada. AB - Winning the 2000 International River prize resulted from a significant investment by the community of Ontario over a long period of time in the Grand River Conservation Authority, GRCA. Innovative partnerships with the business community, government, universities, First Nations tribes, environment groups and the general community have been the hallmark of a broad-based river management program that has returned the Grand River to a healthy environment and usable resource. Recognising this, the Grand River received special recognition from the Canadian Heritage Rivers Board--as a designated Heritage River. Not content with the achievements of the past, the GRCA has turned its attention to the future needs of this river system and to increasing the depth and breadth of community partnerships. PMID- 11419140 TI - Managing the Brisbane River and Moreton Bay: an integrated research/management program to reduce impacts on an Australian estuary. AB - The Brisbane River and Moreton Bay Study, an interdisciplinary study of Moreton Bay and its major tributaries, was initiated to address water quality issues which link sewage and diffuse loading with environmental degradation. Runoff and deposition of fine-grained sediments into Moreton Bay, followed by resuspension, have been linked with increased turbidity and significant loss of seagrass habitat. Sewage-derived nutrient enrichment, particularly nitrogen (N), has been linked to algal blooms by sewage plume maps. Blooms of a marine cyanobacterium, Lyngbya majuscula, in Moreton Bay have resulted in significant impacts on human health (e.g., contact dermatitis) and ecological health (e.g., seagrass loss), and the availability of dissolved iron from acid sulfate soil runoff has been hypothesised. The impacts of catchment activities resulting in runoff of sediments, nutrients and dissolved iron on the health of the Moreton Bay waterways are addressed. The Study, established by 6 local councils in association with two state departments in 1994, forms a regional component of a national and state program to achieve ecologically sustainable use of the waterways by protecting and enhancing their health, while maintaining economic and social development. The Study framework illustrates a unique integrated approach to water quality management whereby scientific research, community participation and the strategy development were done in parallel with each other. This collaborative effort resulted in a water quality management strategy which focuses on the integration of socioeconomic and ecological values of the waterways. This work has led to significant cost savings in infrastructure by providing a clear focus on initiatives towards achieving healthy waterways. The Study's Stage 2 initiatives form the basis for this paper. PMID- 11419141 TI - Australia's environmental flow initiative: filling some knowledge gaps and exposing others. AB - Many of Australia's river systems have been seriously degraded by inappropriate management of regulated flows. Other systems are facing threats from future water resources developments. There is a lack of information available to aid in allocation of environmental flows to rivers in order that they are managed in an ecologically sustainable manner. The Environmental Flows Initiative (EFI) is a major Australia-wide R&D program into environmental flows, funded through the Natural Heritage Trust (NHT), and administered by Environment Australia (EA). The program aims to identify environmental values, undertake targeted research to identify risks to river systems and flow requirements to sustain environmental values, to trial flow management options, and to evaluate these trials. The NHT relies on matching funding provided by the State and Territory authorities, and supports integrative approaches with emphasis on works on-the-ground where possible. While the EFI will close significant knowledge gaps, other gaps remain. Some of these relate to development and validation of rapid assessment techniques, understanding the importance of flow variability and how to define it, manipulation of flows to control alien species, developing a system of prioritising rivers for environmental flows, and enhancing flows with other catchment, channel and floodplain rehabilitation measures. PMID- 11419142 TI - Assessment of changes to saltwater/freshwater habitat from reductions in flow to the Richmond River estuary, Australia. AB - The Australian climate is highly variable and many Australian estuaries lack a seasonal pattern of freshwater flow. During periods of low freshwater inflow, saline waters enter from the ocean through the estuary mouth. These saline waters enter as density currents or as a result of tidal mixing. During periods of high freshwater inflow from the estuary catchment, salt water is flushed towards the estuary mouth. As a consequence, the saline structure of Australian estuaries can be highly variable, depending on the antecedent rainfall. The Richmond River in northern New South Wales in such an estuary. The biota inhabiting estuaries have varying levels of freshwater and saltwater tolerance and reducing the freshwater flow into and along an estuary will favour saltwater species. However, if extractions of freshwater from an estuary are sufficiently high, freshwater habitat could be eliminated entirely for short periods (-one month) but with consequent, and perhaps long-standing, damage to the environment. This contribution describes a new approach to quantify the impact of changes to freshwater flows to the Richmond River estuary. This approach includes a review of hydrological data gathered over the past century and includes a detailed assessment of: changes to the highly variable freshwater inflows and freshwater extraction below the tidal limit; consequent changes to the highly variable saline structure; and the risk to aquatic biota. The crucial factors of magnitude, frequency and duration of short-term intrusions of saline water into freshwater habitat and their impact are quantified by the methods developed. PMID- 11419143 TI - Nitrogen levels in the Tully River--a long-term view. AB - Enhanced run-off of nutrients from agricultural development in catchments bordering the Great Barrier Reef (GBR) is regarded as a potential threat to inshore reefs. In an attempt to better understand the impact of river run-off, the Australian Institute of Marine Science (AIMS) initiated a sampling program to measure nutrient concentrations in rivers draining to the GBR shelf. Our longest data set is from the Tully River in the wet tropics, sampled in collaboration with the Bureau of Sugar Experiment Stations (BSES). After thirteen years, we ask whether there has been any change in river nutrient levels. Water samples were collected at monthly intervals, with additional, wet-season samples. In recent years, a trend of increase in nitrate (at low flow) and particulate nitrogen concentrations has been observed. In this period, a significant increase in agricultural activity has occurred within the Tully basin. The area under sugarcane and bananas has doubled and fertiliser nitrogen use has increased by 130%. Increased concentrations of nitrogen in the river system may be a consequence of this increase in agricultural activity. PMID- 11419144 TI - Color Doppler sonography: its real role in the evaluation of children with highly suspected testicular torsion. AB - The goal of this study was to assess the reliability of color Doppler imaging in boys who presented with acute scrotal pain and borderline clinical findings. The second purpose of the study was to evaluate the potential change in diagnostic accuracy as a result of employing radiological staff with varying levels of experience. Thirty-eight consecutive patients with highly suspected testicular torsion were enrolled in this prospective study. Clinical examination was followed by color Doppler US (7.5-MHz transducer). The staff of radiologists included four residents and three experienced radiologists. All patients underwent surgical exploration. All original reports of residents were reviewed by experienced radiologists and sensitivity with specificity were determined and compared. Original interpretations of color Doppler imaging yielded sensitivity of 77.8% and specificity of 85%, which changed after review to 88.9 and 90%, respectively. Clinical assessment was accurate only in 47.4%. Agreement between original and retrospective color Doppler diagnosis was obtained in 20 of 23 (86.9%) reviewed cases. Color Doppler US may prevent unnecessary surgery in the cases with conclusive normal and increased blood flow. In all other situations scrotal exploration should be performed. PMID- 11419145 TI - The role of isotropic diffusion MRI in children under 2 years of age. AB - Our objective was to determine the contribution of diffusion MR imaging to standard MR imaging in the neuroradiological evaluation of children less than 2 years of age. Echo-planar diffusion MR imaging was added to standard MR exams in 75 consecutive patients under the age of 2 years. Single-shot echo-planar spin echo T2 weighted images (EPSE-T2) were acquired. Isotropic diffusion-weighted images (DWI), attenuation coefficient maps (ACM), and apparent diffusion co efficient (ADC) maps were calculated offline from images obtained with diffusion gradients (b = 1000 s/mm2) in three orthogonal directions. Two neuroradiologists determined if EPSE-T2, DWI, or ACM contributed new information to spin-echo proton density (SE PD) and T2 studies. In 15 of 18 abnormalities detected in 8 patients with symptoms less than 1 week in duration, DWI and/or ACM added information to SE PD and T2. Diffusion sequences detected five new lesions, showed six lesions with greater conspicuity, and identified four lesions with different diffusion character. In patients with symptoms of more than 7 days duration, diffusion studies added no information. Isotropic diffusion MR contributed to lesion detection and characterization in infants when symptoms were less than 1 week in duration. Diffusion MR is useful in patients with leukodystrophies, metabolic disorders, and patients with acute ischemic lesions. PMID- 11419146 TI - Lymphocytic interstitial pneumonia in children with AIDS: high-resolution CT findings. AB - Pulmonary involvement in children with acquired immunodeficiency syndrome (AIDS) represents a wide spectrum of diseases. Among the non-infectious, non-neoplastic affections associated with AIDS, lymphocytic interstitial pneumonia (LIP) is now a well-recognized entity, but its radiological pattern studied with high resolution computed tomography (HRCT) has rarely been described in children. The aim of this study was to illustrate the HRCT spectrum of pulmonary involvement in children with LIP and to evaluate its usefulness in the early diagnosis of this entity. Twelve children with AIDS, aged 3-9 years (mean age 5 years 7 months), underwent chest radiographs and HRCT. A control group of 7 healthy aged-matched children was also studied in the same conditions. Diagnosis of LIP was based on clinical data and HRCT findings. Eight children of 12 had a reticulonodular pattern on chest radiographs. Two children had normal chest films and two children showed peribronchiolar thickening. High-resolution CT displayed micronodules, 1-3 mm in diameter, with a perilymphatic distribution in all patients. High-resolution CT demonstrated also subpleural nodules in children without reticulonodular opacities on chest radiographs. High-resolution CT is able to define a more specific pattern of abnormalities than conventional chest radiographs in children with LIP, allows an earlier and more confident diagnosis and may be useful for the detection of other pathologies associated with AIDS, such as opportunistic infections or superimposed malignancies. PMID- 11419147 TI - Cystic retroperitoneal lymphangioma: treatment by image-guided percutaneous catheter drainage and sclerotherapy. AB - Retroperitoneal cystic lymphangiomas are extremely rare and the majority are symptomatic during childhood. Although benign, they can compress and infiltrate vital structures. Surgery is curative but is associated with a high complication rate. An alternative treatment strategy is image-guided percutaneous catheter drainage of the lymphangioma followed by sclerotherapy. Resolution of a large retroperitoneal cystic lymphangioma in a 4-year-old child treated by this technique is reported. To our knowledge, this technique has not been previously described in this condition and we believe that it offers significant advantages over surgery. PMID- 11419148 TI - Rare pseudotumors of the urinary bladder in childhood. AB - We report two cases of inflammatory pseudotumors of the urinary bladder, one case of a chronic granulomatous pseudotumor (CGT) and one case of a pseudosarcomatous myofibroblastic (fibromyxoid) tumor (PMT). Both tumors resembled malignancies such as rhabdomyosarcomas regarding clinical appearance and imaging findings and represent rare urinary bladder tumors. The imaging findings on unenhanced and contrast-enhanced MRI as well as histological specimen are presented. Final diagnosis was made following elective surgery. Differential diagnosis of urinary bladder tumors as well as the imaging findings of these clinically comparable cases are discussed. Awareness of these benign lesions may prevent patients from inappropriate therapies such as chemotherapy or radiation therapy. PMID- 11419149 TI - MRI of the arthritic small joints: comparison of extremity MRI (0.2 T) vs high field MRI (1.5 T). AB - The aim of this study was to compare the diagnostic capabilities of extremity MRI (E-MRI) with high-field MRI in arthritic small joints, and to evaluate the patients' acceptance and perceptions of the two MR systems. One hundred three patients (group 1 = 28 patients with RA < 3 years, group 2 = 25 patients with reactive and psoriatic arthritis and mixed connective tissue disease, group 3 = 25 patients with rheumatoid arthritis (RA) more than 3 years and group 4 = 25 patients with arthralgia) underwent dedicated E-MRI and high-field MRI of the wrist and finger joints. Coronal short tau inversion recovery and transversal 3D T1-weighted images before and after gadodiamide (Gd) were performed in both cases to outline the volume of the synovial membrane (Vsm) and to evaluate joints with enhancement, effusion, bone edema, and erosions. Investigators blinded to the clinical findings evaluated the images. Patients' compliance and acceptance of E MRI and high-field MRI were evaluated. The median Vsm obtained on E-MRI did not differ significantly from that obtained on high-field MRI. Vsm = 1 ml (E-MRI) and 1.1 ml (high-field MRI) before Gd and Vsm = 0.1 ml (E-MRI) and 0 ml (high-field MRI) after Gd (Wilcoxon test, p > 0.05). The difference in agreement was 8% for joint enhancement, 2% for joint effusion, 3% for bone edema, and 4% for bone erosions. Of the patients, 64% preferred E-MRI due to more comfortable positioning and less claustrophobia and noise. Extremity MRI of the small arthritic joints is comparable to high-field MRI and more readily accepted than high-field MRI by this patient group. PMID- 11419150 TI - Diagnostic utility of an echo-contrast agent in patients with synovitis using power Doppler ultrasound: a preliminary study with comparison to contrast enhanced MRI. AB - The purpose of this study was to first evaluate Levovist (Schering, Berlin, Germany), an echo-contrast agent, during power Doppler sonography (PDS) in patients with synovitis using asymptomatic joints as controls. Then we evaluated the accuracy of this technique against contrast-enhanced MRI. Forty patients (19 men and 21 women; mean age 40 years) were enrolled on the basis of clinical signs, laboratory tests, and radiographic findings positive for articular inflammatory disease. They were examined with conventional ultrasonography (US) and PDS techniques before and after intravenous contrast medium injection. Fourteen patients then underwent MRI with and without contrast medium 8-14 days after PDS studies. Three expert readers independently evaluated each examination. After contrast medium, synovium in inflammatory arthritis enhanced on PDS compared with normal joints in the same patient. Power Doppler sonography after contrast medium and MRI were concordant in all cases. Power Doppler sonography with contrast medium showed a qualitative increase in signal from synovial vessels, the first sign of synovial changes in inflammatory diseases. PMID- 11419151 TI - CT-guided obturator nerve block for diagnosis and treatment of painful conditions of the hip. AB - Obturator nerve blocks (ONB) have been performed by anaesthesiologists mainly to eliminate the obturator reflex during transurethral resections. An effect on hip pain has also been described. However, being a time-consuming and operator dependent procedure if performed manually, it has not been widely used for chronic hip pain. The purpose of this pilot study was to check whether CT guidance could improve reproducibility of the block (= immediate effect) and to test its potential value for treatment of chronic hip pain. Fifteen chronically ill patients with osteoarthritis underwent a single ONB. Sixteen millilitres of Lidocaine 1% mixed with 2 ml Iopramide was injected into the obturator canal. The patients were followed up to 9 months after the intervention. With a single injection pain relief was achieved for 1-8 weeks in 7 of 15 patients. Excellent pain relief for 3-11 months was achieved in another 4 patients. Reasons for a mid term or even long-term effect based on a single injection of local anaesthetic are not exactly known. The CT-guided ONB is a fast, easy and safe procedure that may be useful for mid-term (weeks) and sometimes even long-term (months) treatment of hip pain. PMID- 11419152 TI - Low-grade chondrosarcoma vs enchondroma: challenges in diagnosis and management. AB - A 28-year-old man presented with a swelling at the right thoracic wall. Computed tomography showed an aggressive process involving the cortex of the rib with concomitant soft tissue mass. However, a needle biopsy specimen revealed an enchondroma and consequently the physician decided to apply a "wait-and-see" strategy. After 3 years of careful follow-up by MR imaging, the patient complained of subtle enlargement of the lesion, which was later confirmed on repeated CT scan. Despite an aggressive appearance on control MR imaging, histopathological examination after incisional biopsy could not differentiate between enchondroma and low-grade chondrosarcoma. Wide excision including previous biopsy trajectory was performed. Diagnosis of a low-grade (grade I) chondrosarcoma was made on findings of the excisional specimen and seeding of cartilage tissue along the previous incisional biopsy trajectory was found. PMID- 11419153 TI - Thyroid acropachy: correlation of imaging and pathology. AB - Thyroid acropachy is a rare manifestation of autoimmune thyroid disease, in the form of soft tissue swelling of the hands and feet with insidious onset, associated with clubbing and characteristic periosteal reactions. It is usually part of a syndrome consisting of a typical triad of thyroid acropachy, exophthalmos, and pretibial myxedema. The purpose of this case report is to demonstrate the imaging features of this typical triad in a 65-year-old-woman. This case is the first in which the MRI features of thyroid dermopathy are documented. PMID- 11419154 TI - MRI of neurologic complications in end-stage renal failure patients on hemodialysis: pictorial review. AB - End-stage renal disease patients who have been on long-term hemodialysis tend to develop central nervous system complications. The most common neurologic complications in this patient group include white matter changes, cerebral atrophy, osmotic demyelination syndrome, dialysis encephalopathy, hypertensive encephalopathy, intracranial hemorrhage, infarct, sinus thrombosis, and infection. Clinical evaluation of these patients is somehow complicated and MRI is important before establishment of the therapy. The purpose of this article is to illustrate the range of MRI findings of neurologic complications in end-stage renal failure patients on hemodialysis with etiologic factors. PMID- 11419155 TI - Micropituitarism and cortical dysplasia: an unknown association of two uncommon CNS disorders. AB - We describe a case of two known pathologies of the CNS in an unusual association: the concomitant presentation of the micropituitarism and cortical dysplasia. To our knowledge, this association is unreported to date. PMID- 11419156 TI - Hemangiopericytoma of the parotid gland: CT and MR features. AB - Hemangiopericytomas are uncommon vascular neoplasms with rare occurrence in the head and neck region. They originate from the pericytes, which are small, oval cells encircling capillaries. Hemangiopericytomas traditionally appear in the retroperitoneum and in the capillaries of the extremities. A case of hemangiopericytoma of the parotid gland is presented. The clinical, surgical, histologic, and radiologic features are described and discussed. PMID- 11419157 TI - Skull base granuloma: an unusual location of idiopathic midline destructive disease. AB - A rare case of idiopathic midline destructive disease is presented, which is an entity of the so-called midline granuloma syndrome. Differentiation from other granulomas, especially from Wegener's granulomatosis, is important. This report shows the MR findings in a patient with atypical clinical presentation of histopathologically proven idiopathic midline destructive disease. PMID- 11419158 TI - MR imaging of primary malignant lymphoma in the larynx. AB - We present MR findings in a patient with primary laryngeal lymphoma. The MR images showed a homogeneous mass in the right supraglottic larynx extending to the true vocal cord through the paraglottic space, of which signal intensity was intermediate both on T1- and T2-weighted images. The tumor was moderately enhanced and preserved mucosal layers were demonstrated as hyperintense bands on gadolinium-enhanced fat-suppressed T1-weighted images. Primary laryngeal lymphoma should be included in the differential diagnosis for a homogeneous solid mass in the supraglottic submucosal area. PMID- 11419160 TI - Online first publication. PMID- 11419159 TI - Experimental and theoretical assessment of the performance of Gd2O2S:Tb and La2O2S:Tb phosphors and Gd2O2S:Tb-La2O2S:Tb mixtures for X-ray imaging. AB - The purpose of this work was to investigate and compare the imaging performance of Gd2O2S:Tb and La2O2S:Tb phosphors as well as of Gd2O2S:Tb and La2O2S:Tb mixtures for use in X-ray imaging detectors (intensifying screens). Phosphors were supplied in powder form and were used to prepare test screens. Three types of screens were prepared: Gd2O2S:Tb; (Gd50La50)O2S:Tb; and La2O2S:Tb. Screens were excited by X-rays with tube voltages from 40 to 120 kV and their efficiency (light intensity or light energy flux over exposure) was measured with a photomultiplier and a dosimeter. The light spectrum was also measured with a monochromator. From these measurements, the number of emitted photons per incident X-ray (NEP) and the zero frequency detective quantum efficiency (DQE(0)) of the screens were determined. Additionally, modulation transfer function (MTF) was measured by the square wave response function (SWRF) method. A theoretical model calculating NEP and DQE was also developed to fit experimental data and predict the performance of Gd2O2S:Tb-La2O2S:Tb mixtures by weight from 10-90% to 90-10%. Gd2O2S:Tb screens exhibited highest NEP, DQE, and MTF at tube voltages higher than 55 kV and lower than 45 kV, whereas La2O2S:Tb screens had better NEP, DQE, and MTF within the 45- to 55-kV range. Maximum NEP values were higher than 700 at 100-120 kV while maximum DQE(0) was 0.314 at 80 kV. Gd2O2S:Tb screens are more efficient for high X-ray tube voltage applications (e.g., abdominal imaging, chest radiography, lumbar spine radiography, CT) and for very low voltage applications (e.g., mammography). La2O2S:Tb screens are useful for medium-range X ray voltages (e.g., pediatric radiography). PMID- 11419161 TI - MR enteroclysis protocol optimization: comparison between 3D FLASH with fat saturation after intravenous gadolinium injection and true FISP sequences. AB - The aim of this study was to introduce the true fast imaging with steady-state precession (FISP) sequence for MR enteroclysis and compare it with the already used T1-weighted fast low-angle shot (FLASH) sequence. Twenty-one patients underwent both MR and conventional enteroclysis. The MR enteroclysis examination was performed after administration of an iso-osmotic water solution through a nasojejunal catheter and the following sequences were included: (a) true FISP; and (b) 3D FLASH with fat saturation after intravenous injection of 20 mg Buscopan or 1 mg glucagon and 0.1 mmol/kg gadolinium chelates. The true FISP sequence provided images with significantly fewer motion artifacts, whereas 3D FLASH was less sensitive to susceptibility and chemical shift artifacts. The homogeneity of endoluminal opacification, wall conspicuity, and distention of the small bowel were very good to excellent and the two sequences presented no statistically significant differences here. True FISP provided significantly better overall image quality than did 3D FLASH. The true FISP sequence can provide good anatomic demonstration of the small bowel on T2-like images and could be combined with T1-weighted FLASH images for an integrated protocol of MR enteroclysis. PMID- 11419162 TI - Percutaneous radiofrequency ablation of small hepatocellular carcinoma: long-term results. AB - The aim of this study was to evaluate the effectiveness and the safety of percutaneous radiofrequency (RF) thermal ablation of hepatocellular carcinoma (HCC) in 88 patients with a long follow-up, and to compare conventional electrodes and expandable electrodes. Eighty-eight patients with 101 hepatocellular carcinoma nodules (< or = 3.5 cm in diameter) underwent RF thermal ablation by means of either conventional electrodes or an expandable electrode. Therapeutic efficacy was evaluated with dynamic contrast CT, serum alpha-feto protein level, US examination at the end of the treatment, and during follow-up. Complete necrosis was obtained in all tumor nodules in a mean number of 3.3 sessions (tumor treated by conventional electrodes) or 1.5 sessions (tumor treated by expandable electrode). The mean follow-up was 34 months; overall survival rate was 33% at 5 years. Disease-free survival at 5 years was 3%; local recurrence rate was 29% in patients treated with conventional electrodes; 14% in patients treated with the expandable electrode. Two major complications and 14 minor complications were observed. Radiofrequency thermal ablation in small HCC is very effective with a low percentage of major complications. The use of an expandable electrode substantially reduced the number of treatment sessions but did not modify the overall survival rate and the disease-free survival rate. PMID- 11419163 TI - Peripancreatic fat necrosis mimicking pancreatic cancer. AB - A case of peripancreatic fat necrosis, after an episode of acute pancreatitis, which mimicked pancreatic cancer with lymph node metastases, is presented. We describe the imaging findings with helical CT scanning and with unenhanced and mangafodipir-enhanced MR imaging, with special emphasis on the differential diagnoses. PMID- 11419164 TI - Unusual presentation of a pancreatic insulinoma in helical CT and dynamic contrast-enhanced MR imaging: case report. AB - Insulinomas are pancreatic neoplasms that can be radiologically characterized typically because of their tendency to present intense and early contrast enhancement with a wash-out phenomenon. In this sense, we report an unusual case of a hypovascular solid pancreatic insulinoma confirmed with surgery and pathologic analysis, in a patient with normal serum insulin levels. In the two phase helical CT, the mass behaved as a hypodense lesion with respect to the surrounding pancreatic parenchyma during the arterial phase and as a hypointense lesion during the dynamic contrast-enhanced MR imaging. Pathologic examination demonstrated a hypercellular tumor with poor vascularization of intervening stroma which showed prominent amyloid deposits. PMID- 11419165 TI - What is left of i.v. urography? AB - Since its introduction into clinical practice in the early 1930s, intravenous urography (IVU) was the primary imaging technique for the investigation of urinary system disorders for many years, until the advent of digital cross sectional-imaging techniques gradually started to undermine many of its indications. Intravenous urography has been superseded for some indications such as renovascular arterial hypertension, prostatic dysuria, renal failure, palpable abdominal masses and recurrent urinary tract infection in women. Intravenous urography has been reduced, in the sense that it is no longer a primary examination, for other clinical indications such as renal colic, renal trauma, uroseptic fever, asymptomatic haematuria, medical haematuria, obstructive uropathies and follow-up of various disorders. Intravenous urography is indicated and often mandatory in congenital anomalies of the urinary tract, prior to endourological procedures, possible fistulas, renal transplantation, tuberculosis and ureteral pathology. In conclusion, IVU is still the examination of choice where there is a need to visualize the entire urinary system and to evaluate the state of the papillae and calyces. Computed tomography urography and MR urography are the imaging modalities ready in the near future to replace IVU. PMID- 11419166 TI - Imaging of retroperitoneal ganglioneuroma. AB - The aim of this study was to describe the radiological appearance of retroperitoneal ganglioneuroma. We retrospectively reviewed seven cases of histologically proven retroperitoneal ganglioneuroma. Ultrasound and enhanced CT were obtained in all cases, and MRI in three cases. The masses were well circumscribed, ranged in size from 5 x 3 x 3 to 10 x 6 x 4 cm. In three cases close relationships between the tumor mass and major blood vessels were noted, resulting in vessel displacement or surrounding, but without compression or occlusion. On ultrasound examination the tumor showed a heterogeneous solid echostructure. Non-enhanced CT showed homogeneous or mildly heterogeneous low attenuation, and a punctate calcification was seen in one case. Contrast uptake was absent (n = 1) or delayed (n = 6). Progressive but incomplete enhancement was observed in three cases. On MRI, T2-weighted images showed a high signal intensity. Dynamic studies depicted the same enhancement pattern as described on CT. Ganglioneuroma is a rare tumor which should nevertheless be included in differential diagnosis of retroperitoneal masses when presenting as a well delimited tumor with possible tendency to surround or displace major blood vessels, low density on non-enhanced CT, and delayed progressive enhancement on CT and MRI. PMID- 11419167 TI - MR imaging of clear cell carcinoma of the ovary. AB - Magnetic resonance imaging findings are reported for 12 pathologically proven lesions of clear cell carcinoma (CCC) of the ovary in 11 women (mean age 50 years). T1- and T2-weighted MR images were obtained in all patients, and gadolinium-enhanced MR images were obtained in 9. The mean diameter of the tumors was 13 cm. Seven patients presented with stage-I tumors. All 12 lesions consisted of cystic masses with solid protrusions occurring in 10 and solid masses in 2. The cysts were unilocular in 9 lesions and multilocular in 1. In four lesions, the cysts displayed with high intensity on T1-weighted images. Round solid protrusions were identified in 8 lesions. In 5 lesions, the number of protrusions was only a few. The solid portions of 5 masses had slightly high-intensity regions on T1-weighted images. The number of patients with ascites was three. Magnetic resonance imaging of CCC usually shows a unilocular large cyst with solid protrusions, which are often round and few in number. Such MR imaging findings suggest malignant tumor but are not specific. PMID- 11419168 TI - Imaging of membranous dysmenorrhea. AB - Membranous dysmenorrhea is an unusual clinical entity. It is characterized by the expulsion of huge fragments of endometrium during the menses, favored by hormonal abnormality or drug intake. This report describes a case with clinical, US, and MRI findings before the expulsion. Differential diagnoses are discussed. PMID- 11419169 TI - Persistent mullerian duct syndrome with teratoma in an ectopic testis: imaging features. AB - The persistent mullerian duct syndrome represents a rare form of male pseudohermaphroditism, secondary to mullerian inhibiting factor (MIF) deficiency. We describe imaging findings in a 30-year-old male (46 XY karyotype) with bilateral cryptorchidism and mullerian duct anomalies (presence of uterus and fallopian tubes). Grade-III teratoma with yolk sac tumour was detected in one of the undescended testis, lying in the pelvic cavity. The other testis was in the inguinal canal. The rest of the wolffian duct structures (e.g. prostate, seminal vesicles) were nearly normal. Very few reports of imaging findings of this entity have been published thus far, probably because of the rarity of entity, incidental detection of most of the cases at surgery and relatively asymptomatic clinical presentation. PMID- 11419170 TI - Non-tuberculous iliopsoas abscess due to perforated diverticulitis presenting with intestinal obstruction and a groin mass. AB - Psoas abscess is an uncommon condition and, contrary to traditional teaching, tends to be of non-tuberculous aetiology in developed countries. Diagnosis can be delayed since presenting features are non-specific and in many instances misleading, necessitating a high degree of clinical suspicion and early resort to cross-sectional imaging using CT or MRI. We present a case of iliopsoas abscess secondary to perforated diverticulitis to illustrate the difficulty encountered in early diagnosis and to show that successful management of secondary psoas abscess necessitates surgical resection of the underlying condition in most cases. PMID- 11419171 TI - Preoperative arterial embolization in heterotopic ossification: a case report. AB - We report a case of preoperative embolization in a 64-year-old patient suffering from total stiffness of the right hip joint due to heterotopic ossification following brain injury and pertrochanteric fracture of the right femur. A previous attempt of operative treatment could not be performed successfully due to bleeding complications. After the embolization of the correlating hypervascularisation, the surgical procedure was redone and finished with good result and minimal bleeding complications during the operation and a tolerable drop of the haemoglobin concentration postoperatively. PMID- 11419172 TI - Gadolinium-enhanced MR angiography of the breast: is breast cancer associated with ipsilateral higher vascularity? AB - The aim of this study was to assess a possible association between breast malignancy and ipsilateral higher vascularity on gadolinium-enhanced MR angiography. One hundred six patients were examined by dynamic gadolinium enhanced 3D MR imaging. Magnetic resonance angiographic views were generated by image subtraction and maximum intensity projection. The study included 85 patients with unilateral malignant breast neoplasms and 21 with unilateral benign lesions. Three blinded readers independently reviewed the MR angiograms after masking the lesions and the corresponding contralateral sites. The readers were asked to determine whether vascularity was higher on the right side, higher on the left side, or equal on both sides. The results were analyzed by the Kappa statistic and Pearson's chi-square test. The blood vessels of the breasts were clearly seen in all cases. There was good agreement among the observers (kappa > 0.54) in assessing vascularity on both sides. Breasts harboring malignant neoplasms were found to have a higher vascularity than the contralateral breasts (p < 0.005). This sign of malignancy had a sensitivity of 76.5%, a specificity of 57%, and an accuracy of 72.6%. Blood vessels of the breast can be depicted by MR angiography. Unilateral malignant neoplasms are associated with a higher ipsilateral vascularity. In conjunction with other indications of malignancy on gadolinium-enhanced MR images, a higher ipsilateral vascularity may serve as an additional sign of malignancy. PMID- 11419173 TI - Contrast-enhanced color Doppler ultrasound characteristics in hypervascular breast tumors: comparison with MRI. AB - The aim of this study was to evaluate the accuracy of contrast-enhanced color Doppler ultrasound (CE-US) in comparison with contrast-enhanced MR imaging (CE MRI) in the discrimination of hypervascularized breast tumors. An additional CE US of the breast was preoperatively performed in 40 patients with a hypervascular breast lesion detected on CE-MRI. The presence of blood flow signals and the morphological characteristics of the vessels in the breast lesions were evaluated pre- and post-contrast administration, as well as the dynamic aspects of the Doppler signal, including time interval to maximum signal enhancement and persistence of the signal enhancement. Twenty-three carcinomas and 17 fibroadenomas were explored. Considering initial signal enhancement > 100% after the administration of contrast material as a criterion suggesting malignancy, CE MRI showed a sensitivity of 100% and a specificity of 76.5% in the detection of malignant breast tumors. Color Doppler signals were consistently demonstrated in all carcinomas and in 68.7% of fibroadenomas after the administration of Levovist, with CE-US showing a sensitivity of 95.6% and a specificity of 5.9%. Neither the mean number of vessels per tumor, nor the location of vessels, the time to maximum increase of the Doppler signal or the persistence of signal enhancement showed significant differences between benign and malignant lesions. Additional CE-US does not increase the low specificity of MRI in patients with hypervascularized breast tumors. PMID- 11419174 TI - Stereotactic breast biopsies: analysis of needle deviation based on stereotactic views. AB - Stereotaxy is widely used for breast biopsy and needle localization of mammographically detected lesions. If a lesion shift occurs during stereotaxy, corrections (even though possible with vacuum biopsy) is difficult due to the difficult assessment of the exact 3D shift. In this study we investigated the correlation between lesion shift (in up to three dimensions) and its visualisation on the stereotactic images (0 degree, -15 degrees, +15 degrees). The study was performed on a Fischer prone table (Fischer Imaging Europe, Vejle, Denmark) using a 3.8-mm steel ball (as lesion) and a 20-G needle. The 17 major malpositions of the ball with respect to the needle were imitated and imaged. A simple rule is suggested as to how the deviation in all three dimensions can be detected from the scout and the two stereotactic views. The rule proved to be a valuable tool to correctly assess lesion shifts. PMID- 11419175 TI - Paired inspiratory/expiratory spiral CT and continuous respiration cine CT in the diagnosis of tracheal instability. AB - In tracheo- and bronchomalacia, localisation and determination of collapse is necessary for planning of surgical procedure. We compared inspiratory and expiratory spiral CT, cine CT, bronchoscopy, exemplary cine MR, and evaluated the clinical relevance. Twenty-nine patients (2 follow-ups; mean age 61 years, age range 27-85 years) with suspected or verified tracheal stenosis or collapse (post tracheotomy: n = 17; neoplasm: n = 5; other: n = 7) underwent paired breath-hold inspiratory and expiratory spiral CT. Forty-five additional cine CT were performed at 1-4 levels (mean 1.5) during continuous respiration (increment 100 ms) to clarify respiratory collapse. The tracheal cross-sectional diameters of both techniques were calculated. Comparison with bronchoscopy, follow-up, and influence upon therapy were evaluated retrospectively. Exemplary comparison with cine MR (8 frames/s) was done in 3 cases. In addition to bronchoscopy, further information concerning localisation, extent, collapse, stability of the tracheal wall, distal portions of the stenosis and extraluminal compressions were obtained. A significantly higher degree and more pathological collapses (> 50%) were seen using cine CT (38%) compared with paired spiral CT (13%; degree: p < 0.0001; number: p < 0.001). The findings changed the further therapeutic procedure in 16 of 29 patients. Further stenoses were excluded and bronchoscopy was verified in another 13 of 29. Temporal resolution of cine CT and cine MR is sufficient; however, spatial resolution of cine MR is inferior. Paired inspiratory and expiratory spiral CT localises tracheal stenoses and demonstrates relevant extraluminal compression. Significantly improved evaluation of respiratory collapse and further information of localised tracheal instability is obtained by cine CT. Cine MR promises more functional information especially due to free choice of imaging plane. PMID- 11419176 TI - Herpes simplex virus 1 pneumonia: conventional chest radiograph pattern. AB - The aim of this study was to describe the findings on plain chest radiographs in patients with herpes simplex virus pneumonia (HSVP). The study was based on 17 patients who at a retrospective search have been found to have a monoinfection with herpes simplex virus. The diagnosis was established by isolation of the virus from material obtained during fiberoptic bronchoscopy (FOB) which also included broncho-alveolar lavage and tissue sampling. Fourteen patients had a chest radiograph performed within 24 h of the date of the FOB. Two radiographs showed no abnormalities of the lung parenchyma. The radiographs of the other 12 patients showed lung opacification, predominantly lobar or more extensive and always bilateral. Most patients presented with a mixed airspace and interstitial pattern of opacities, but 11 of 14 showed at least an airspace consolidation. Lobar, segmental, or subsegmental atelectasis was present in 7 patients, and unilateral or bilateral pleural effusion in 8 patients, but only in 1 patient was it a large amount. In contradiction to the literature which reports a high correlation between HSVP and acute respiratory distress syndrome (ARDS), 11 of 14 patients did not meet the pathophysiological criteria for ARDS. The radiologist may suggest the diagnosis of HSVP when bilateral airspace consolidation or mixed opacities appear in a susceptible group of patients who are not thought to have ARDS or pulmonary edema. The definite diagnosis of HSV pneumonia can be established only on the basis of culture of material obtained by broncho-alveolar lavage. PMID- 11419177 TI - Blunt traumatic rupture of the pericardium with cardiac herniation: two cases diagnosed using computed tomography. AB - Traumatic ruptures of the pericardium with cardiac herniation are infrequent, and their radiological pattern little familiar, so that they are often missed preoperatively. Few reports have emphasised the use of a CT scan as a tool for diagnosis and CT scan signs have not been well documented. We report on two cases of traumatic herniation of the heart for which a CT scan brought a major contribution for diagnosis. We describe the presence of an empty pericardial sac on CT slices which allowed us to diagnose the cardiac herniation. These observations demonstrate that CT scans can contribute to the diagnosis of pericardial rupture with cardiac herniation. PMID- 11419178 TI - Gamma/delta T cells provide innate immunity against renal cell carcinoma. AB - Host immune function plays a certain role against the development of renal cell carcinomas (RCCs), but the mechanism is not entirely understood. Human gamma/delta (gamma/delta) T cells defend the body against infection. In this study, we clarify the role of gamma/delta T cells in the surveillance system against RCCs by analyzing the gamma/delta T cells in peripheral blood mononucleocytes (PBMs) and tumor infiltrating lymphocytes (TILs) from 41 patients with RCCs. The results showed that the number of gamma/delta T cells expressing V gamma 2 and V delta 2 in variable elements of TCR was elevated in the PBMs in 10 patients, but not in any of 32 healthy individuals. The proportion of patients with an elevated number of gamma/delta T cells (> 10%) increased with cancer stage. The level of the gamma/delta T cells decreased after surgery. The gamma/delta T cells in the TILs were more activated than those in the PBMs. Evaluation of the junctional diversity of TCR V gamma 2 and V delta 2 chains showed that the increased peripheral blood gamma/delta T cells were oligoclonal rather than polyclonal. Taken together, our findings suggest that gamma/delta T cells recognize certain RCC-related antigens and play a role in the surveillance system against RCCs. PMID- 11419179 TI - The anti-tumor activity of anti-CTLA-4 is mediated through its induction of IFN gamma. AB - The T-cell-specific receptor, CTLA-4, has been demonstrated to be a potent negative regulator of lymphocyte activation, the functional significance of which has been demonstrated in murine tumor models using blocking antibodies. However, the mechanism(s) involved in enhancing tumor regression has not been identified. In this study, we determined whether IFN gamma was playing a role in this activity. In vitro, anti-CTLA-4 enhanced IFN gamma production by lymph node cells obtained from tumor-bearing mice (351 pg/ml vs 77 pg/ml). Additionally, fibrosarcoma-challenged animals treated with anti-CTLA-4 had elevated levels of the IFN-inducible enzyme 2-5-OAS in draining lymph nodes (850 pM vs 260 pM for controls) and an increased amount of IFN gamma in tumor lysates (at day 7, 620 pg/100 micrograms vs 160 pg/100 micrograms in controls). The importance of IFN gamma was demonstrated by the ability of neutralizing antibodies to completely abrogate the anti-tumor effects of anti-CTLA-4. Moreover, fibrosarcoma cells were shown to be exquisitely sensitive to IFN gamma-mediated class I upregulation and histological examination of tumors from anti-CTLA-4-treated mice revealed a trend toward increased tumor cell apoptosis and decreased angiogenesis. These studies have demonstrated that one mechanism for the anti-tumor effects of anti-CTLA-4 relates to its ability to augment IFN gamma production, resulting in an increased expression of class I on the tumor, enhanced apoptosis, and a decrease in blood vessel growth. PMID- 11419180 TI - High-scale expansion of melanoma-reactive TIL by a polyclonal stimulus: predictability and relation with disease advancement. AB - The rationale of treating melanoma patients by infusion with tumor-infiltrating leukocytes (TIL) is to perform an adoptive therapy through injection of tumor specific T cells. Nonetheless, methods currently used for ex vivo TIL expansion have not been evaluated for their efficacy to expand TAA-specific T cells. We have addressed this question here, using a culture method in which high TIL growth was induced by a polyclonal T cell stimulus. Intracellular cytokine assays were performed to measure the proportion of T cells responding to autologous tumor cells among the lymphocytes from lymph node biopsies (TIL) of 26 patients with stage III melanoma. The data show that TIL from 18 of these patients contained detectable amounts of tumor-specific T cells before expansion. Although they decreased somewhat in percent abundance during expansion, they were still present afterwards, ranging from 0.3 to 13.8%. Since a median number of 1.7 x 10(10) TIL was obtained from these patients (starting from 3.6 x 10(6) TIL), a total amount of tumor-reactive cytokine-secreting TIL of between 2.8 x 10(6) and 1.12 x 10(9) was obtained in each case from 18 patients. The TIL populations from 8 patients did not contain tumor-reactive T cells: neither before expansion, nor after expansion. Lack of tumor-reactive TIL only occurs for patients bearing several tumor-invaded lymph nodes (40%), but not for those having a single invaded lymph node. Therefore, high numbers of tumor-reactive T cells can be produced, through a polyclonal TIL stimulation, from most early stage III melanoma patients but from only about half of the patients with a more disseminated disease. For this last group, the possibility of getting tumor reactive TIL can be predicted by checking the presence of these cells before expansion. PMID- 11419181 TI - Anti-self antibodies selected from a human IgD heavy chain repertoire: a novel approach to generate therapeutic human antibodies against tumor-associated differentiation antigens. AB - Human antibodies were isolated by phage display from a naturally expressed human antibody repertoire. Antibody selection was carried out against the epithelial cell adhesion molecule (EpCAM) or 17-1A antigen, that in a clinical trial had been successfully used as a target for antibody therapy of minimal residual colorectal cancer. VH chains were selected from the human IgD repertoire expressed on naive B2 and autoreactive B1 lymphocytes. By guiding the selection through a murine template antibody, two EpCAM-specific human antibodies, HD69 and HD70, were obtained that closely resembled the murine therapeutic 17-1A antibody in their binding properties when expressed as complete huIgG1 molecules in CHO cells. However, both human antibodies recruited human cytotoxic effector cells far more efficiently than the murine 17-1A antibody used for clinical trials. Therefore, and in view of the long in vivo half-life of human IgG1 antibodies, HD69 and HD70 are regarded as highly promising third generation versions of the murine therapeutic antibody. Because of their origin from an evolutionary conserved germline VH repertoire, they are expected to exhibit minimal immunogenicity in patients. PMID- 11419182 TI - Infiltration by inflammatory cells required for solar-simulated ultraviolet radiation enhancement of skin tumor growth. AB - In this study we compared the effects of subinflammatory and inflammatory doses of solar-simulated ultraviolet (UV) radiation on enhancement of skin tumor growth, sensitization to haptens and cellular changes within the epidermis of C3H/HeN mice. Tumors transplanted into mice 3 days after exposure to inflammatory, but not subinflammatory, doses of UV radiation had a higher growth rate than those tumors inoculated into unirradiated control mice. Both doses of UV radiation suppressed the induction of contact hypersensitivity and induced tolerance when hapten was painted onto the skin 3 days after irradiation. Skin exposed to the higher, but not the lower, dose of UV radiation contained significantly increased numbers of CD11b+, CD45+ MHC class II- and CD45+ MHC class II(hi) inflammatory cells 3 days post-irradiation. The immunosuppression correlated with a reduction in Langerhans cells and dendritic epidermal T cells. Collectively, this suggests that suppression to contact sensitizers is due to the UV radiation effects on Langerhans cells and dendritic epidermal T cells. While these effects may also suppress the induction of anti-tumor immunity, at higher doses of UV radiation inflammatory cells may enhance tumor growth by a non immunological mechanism. PMID- 11419183 TI - Lymphocyte subsets and viral load in patients with HIV-associated non-Hodgkin's lymphoma treated with anti-CD20 monoclonal antibody and chemotherapy. AB - The anti-CD20 monoclonal antibody Rituximab is a novel antitumor agent used in association with chemotherapy (CT) for the treatment of high-grade/intermediate non-Hodgkin's lymphomas (NHL) in HIV-negative populations. This therapeutic combination is currently also being explored in HIV-positive patients with NHL (HIV-NHL). The objective of our study was to determine CD4 and CD8T cell counts, HIV plasma viremia and proviral load in patients with CD20-positive HIV-NHL treated with Rituximab plus CT and highly active antiretroviral therapy (HAART). We studied eight patients with HIV-NHL treated by anti-CD20 and CT before, after three, and after six cycles of therapy; CD4, CD8 and CD19 lymphocyte subsets were measured by monoclonal antibodies and flow cytometry. HIV plasma viremia was determined by the b-DNA assay, and proviral load by a quantitative competitive PCR. CD4T cell counts remained stable after three cycles of therapy, while a significant reduction of this subset was present at the end of therapy. HIV plasma viremia was significantly reduced after the third cycle, but returned to pretreatment levels at the end of therapy; we also observed individual fluctuations of proviral load during therapy, this marker being increased in two out of three patients at the end of therapy. These observations suggest that Rituximab plus CT accelerated the rate of CD4 depletion and of HIV replication in the peripheral blood of HIV-NHL patients and that HAART may be able to delay these effects. PMID- 11419184 TI - Tumor cell killing by in vitro affinity-matured recombinant human monoclonal antibodies. AB - We have developed a method that allows the rapid improvement of the affinity of phage-displayed antibody fragments by selection on intact eukaryotic cells. A single chain Fv fragment, specific for the tumor-associated Ep-Cam molecule, was mutagenized by shuffling of the immunoglobulin light chain variable region and DNA shuffling of both heavy and light chain variable regions. Higher-affinity mutants were selected from small phage display libraries by cell panning under stringent conditions. When converted to an intact fully human antibody, the mutagenized anti-tumor monoclonal antibody displayed an affinity of 0.4 nM, a 15 fold improvement over the affinity of the original antibody. Compared to previously reported affinity maturation schemes, panning on intact cells does not require purified targets for selection and may be particularly useful when the target molecule can not be expressed as a recombinant molecule or easily purified without disrupting its native configuration. In vitro tumor cell killing assays demonstrated an improved performance of the higher-affinity antibody in complement-mediated tumor cell killing. In contrast, the lower-affinity antibody performed somewhat better in antibody-dependent cellular cytotoxicity assays and penetrated better in multicell spheroids of tumor cells, an in vitro model for the tumor penetration capacity of antibodies. PMID- 11419185 TI - Address of Christine Grant, Commissioner, Department of Health and Senior Services. PMID- 11419186 TI - Medical research, patients' rights, and modernizing Medicare are health care priorities. PMID- 11419187 TI - Radiation during pregnancy. AB - Radiation exposure to pregnant women from diagnostic procedures will almost never result in fetal-absorbed doses that are high enough to cause damage to the fetus unless multiple examinations are performed. Prenatal radionuclide dose or technical factors of X-ray exposure should be recorded to facilitate estimation of fetal dose. Fetal doses of less than 100 mGy (10 rad) do not increase the incidence of fetal malformation. Doses of 200 to 500 mGy (20 to 50 rad) should be evaluated in the context of the type of radiation and time of exposure relative to fetal development and may be associated with a reduction in IQ. Radiation doses greater than 500 mGy (50 rad) result in a significant risk of growth retardation and CNS damage. PMID- 11419188 TI - Understanding PSA and prostate cancer risk assessment. AB - The specificity of serum PSA in screened populations is relatively low because of its inability to discern benign causes of PSA elevation from occult malignancy. An improved understanding of this widely available serum test and the availability of new modalities, such as endorectal MRI, may allow the early detection and treatment of cancer in those men who have it and may avoid unnecessary repetitive prostate biopsies in men with benign conditions. PMID- 11419189 TI - New Jersey is first state in the nation to use electronic death record system. PMID- 11419190 TI - Professional lability insurance: I. A historical perspective. AB - Medical professional liability insurance (also known as malpractice insurance) is designed specifically to protect physicians, clinics, hospitals and health care facilities, and other health personnel in the event that a patient files a claim against them, alleging negligence, with resulting injury or harm. It is one of the largest expenses for any medical practice and is a high priority concern of every physician. The market for this insurance, the approach to evaluating it, and the cost of it have changed dramatically in the last 20 years. PMID- 11419191 TI - The MSNJ Physicians' Health Program continues to lead the nation. PMID- 11419192 TI - Solar protection in Norman Foster's hands creates an elegant gathering place for scientists at Stanford University's new Center for Clinical Sciences. PMID- 11419193 TI - Boots the Chemist, Nottingham, England. PMID- 11419194 TI - Preserve your work in PDFs. AB - Conveying architectural documents electronically can cause problems with graphic appearance. With Portable Document Files (PDFs), you can combine graphics, text, and even sound and video without losing your looks. PMID- 11419195 TI - Architecture rediscovers being green. PMID- 11419196 TI - Mentoring women physicians. PMID- 11419197 TI - On the front lines of organized medicine. President of ACP-ASIM and Georgia physician--Sandra Adamson Fryhofer, M.D. PMID- 11419198 TI - How to manage teen drinking (the smart way). PMID- 11419199 TI - What do you tell the kids? How a recovering alcoholic plans to urge his daughter to abstain. PMID- 11419200 TI - Herbal warning. PMID- 11419201 TI - Partnerships with "prehospital" teams. PMID- 11419202 TI - Build a Web-centric culture for your physicians. PMID- 11419203 TI - A healing presence. The Catholic health ministry involves the practice of faith in the practice of medicine. PMID- 11419204 TI - The public juridic person in action. An interview with Sr. June L. Ketterer, SGM, and David R. Lincoln of Covenant Health Systems. Interview by Bob Stephens. PMID- 11419205 TI - Mentors and mentoring. Health care workers hope to find integrity in their work, organizations, and leaders. PMID- 11419206 TI - Increasing market share. Second in a series examining revenue growth strategies in a difficult health care market. PMID- 11419207 TI - Caring for retired religious. Congregations have choices in deciding how and where to provide care for aging members. PMID- 11419208 TI - Volunteers and Catholic health care. Auxiliary workers should be full partners in the Catholic health ministry. PMID- 11419209 TI - Community networks. Parish Health Promoter Program/El Programa de Promotores de Salud de la Iglesia, Portland, OR. PMID- 11419210 TI - The making of Ascension Health. The bringing together of two Catholic health care cultures was a complex effort. PMID- 11419211 TI - Creating a mission-based culture. Merging organizations will choose one of four possible approaches. PMID- 11419212 TI - The spirit of the place. Atlanta system combines the "southern tradition" with the "mercy philosophy". PMID- 11419213 TI - Legal issues in mergers and acquisitions. Several noteworthy legal developments have arisen since the last wave of merger activity. PMID- 11419214 TI - The parable of the dance. PMID- 11419215 TI - Public relations lessons from an aborted Catholic/secular merger. PMID- 11419216 TI - Ministry leaders as spiritual leaders. PMID- 11419217 TI - Paradigm lost. PMID- 11419218 TI - Knockoffs on the pharmacy shelf. PMID- 11419219 TI - No safety in the numbers. AIDS rises among young. PMID- 11419220 TI - Don't sour on milk; the health benefit beats any risks. PMID- 11419221 TI - Comprehending speaker intent in rebuttal analogy use: the role of irony mapping, absurdity comparison and argumentative convention. AB - Rebuttal analogies (e.g., "Politicians arguing over the renaming of an airport is like watering your petunias when your house is on fire") are commonly used as responses in verbal conflicts. The following study investigated the role that irony mapping, absurdity comparison, and argumentative convention play in interpreters' derivations of speaker's intentions in using rebuttal analogies. In general, these intentions are to demonstrate the unsoundness of the opposed proposition ("argument") and its advocate ("social attack"), (Whaley & Holloway, 1996). Three rebuttal analogy types were rated on argumentativeness and social attack in verbal conflict and nonverbal conflict scenarios. The results of six experiments on 120 participants found that analogies with ironic bases (e.g., "Doubling the defense budget in order to intimidate North Korea is like using a chainsaw to file your nails") were perceived as more of a social attack and as more argumentative than analogies with absurd (e.g., "... using ketchup to wax your car") and nonironic (e.g., "... using a nailfile to file your nails") bases. No difference was found between the two scenario types. Norming data confirmed equivalence of absurdity of ironic and absurd bases, and greater irony of ironic over absurd bases. The results thus implicate hearers' use of the ironic structure between bases and targets in the interpretation of rebuttal analogies rather than mere absurdity comparison or argumentative convention. PMID- 11419222 TI - Emotion in language and speech: methodological issues in naturalistic approaches. AB - Researchers currently seek to improve validity in speech and language studies by adopting naturalistic procedures In emotion-display research, validity is threatened by standard experimental controls which diminish the naturalism of stimuli and response ranges. We report two experiments comparing the adequacy of naturalistic with standard procedures. Experiment 1 had 158 judges code 89 samples of naturally-occurring emotional speech with free-choice emotion labels, and later with labels from a standard set. When free-choice labels were similar across judges, they were consistent with standard labels, but showed a range of intensity and contextual relevance. We recommend that future studies include wider options for judges when coding emotions. Experiment 2 compared valency ratings of words when presented in, or out of, context. Standard procedures score lexical valencies using affective dictionaries, disregarding natural contexts. Experiment 2 compared 23 judges' valency ratings of words presented individually, and later in their original context. Between 30% and 44% of words were rated differently in context (depending on the statistical significance level adopted). We concluded from Experiment 2 that, where small corpora adequately model a domain, the improved accuracy of valency rating achieved by presenting words in their natural context justifies the extra procedures required. PMID- 11419223 TI - Quantitative characterizations of speech rhythm: syllable-timing in Singapore English. AB - British English and Singapore English are said to differ in rhythmic patterning. British English is commonly described as stress-timed, but Singapore English is claimed to be syllable-timed. In the present paper, we explore the acoustic nature of the suggested cross-varietal difference. In directly comparable samples from British English and Singapore English, two types of acoustic measurements were taken; we calculated a variability index reflecting changes in vowel length over utterances, and measurements reflecting vowel quality. Our findings provide acoustic data which support the hypothesized cross-varietal difference in rhythmic patterning; we show (1) that successive vowel durations are more nearly equal in Singapore English than in British English, and (2) that reduced vowels pattern more peripherally in the F1/F2 formant space in Singapore English than in British English. We complete the paper with a comparison of our vowel variability index with a set of acoustic measures for rhythm proposed by Ramus, Nespor, and Mehler (1999), which focus on variability in vocalic and intervocalic intervals. We conclude that our variability index is more successful in capturing rhythmic differences than Ramus et al. (1999)'s measures, and that an application of our index to Ramus et al.'s intervocalic measure may provide a further diagnostic of rhythmic class. PMID- 11419224 TI - Kinematic event patterns in speech: special problems. AB - The view that each utterance is fundamentally a pattern of serially-ordered events underlies a group of well-known speech kinematic studies emphasizing temporal coordination among articulators. Methodological problems that might affect the validity and significance of conclusions from these studies are identified. Results from a new analysis of synchronous acoustic and fleshpoint kinematic data, recorded from 53 normal young-adult speakers of American English, are then reported. The kinematic data represented speech-related actions of the tongue blade and dorsum, both lips, and the mandible, during the test words special and problem, and were drawn from an existing X-ray microbeam speech production database. Distributions of event patterns across speakers revealed four main results: (1) different patterns for the two test words; (2) a comparable degree of cross-speaker agreement about relative tongue and jaw movement timing, but marked disagreement about lip and jaw movement timing, between test words; (3) highly distinctive movement patterns for some speakers; and, (4) a general conclusion that serial event order, alone, provides very limited understanding of movement patterns produced by individual speakers. By design, these results focus attention on methods of kinematic event pattern analyses, and the general value of such analyses for insights about speech production. PMID- 11419225 TI - Structural computation and the role of morphological markings in the processing of Japanese. AB - In an SOV, strictly head-final language with otherwise relatively free word-order such as Japanese, disambiguating information including verb information is not available until the end of a clause. In such a language, a sentence theoretically has numerous possible syntactic structures before the verb information becomes available. The current study investigated (1) whether syntactic computation takes place in an ambiguous sentence fragment in Japanese and (2) if and how information from constituents other than the verb is utilized. Two experiments were conducted using automated, word-by-word visual presentations with a lexical decision task on an extrasentential target. The results suggest that syntactic computation takes place in Japanese before verb information becomes available. A strong preference was observed for a simplex clause when three differently Case marked arguments belonged to the same clause. The findings also indicate that information from surface Case marking is utilized productively, and when there is ambiguity in marking between arguments and adjuncts, the distinction is made before incorporating the constituents into the existing structure. PMID- 11419226 TI - Generation and recall of alcohol-related information in excessive drinkers: relation to problem severity, outcome expectancies, and stage of change. AB - Generation and recall of information supporting and opposing reduction in alcohol use were examined in 47 excessive drinkers. Generation of reasons to change drinking increased across stages of change and was positively associated with alcohol problem severity, negative alcohol expectancies, benefits of changing drinking, and strength of reasons to change. Generation of reasons not to change drinking was not significantly associated with stage of change but was positively associated with positive alcohol expectancies and strength of reasons not to change. Incidental recall of negative outcomes of changing drinking decreased across stage of change and was positively associated with perceived strength of reasons not to change. Results suggest potential biases in generation and recall of alcohol-related information that could act to maintain excessive drinking by making certain types of information more available during decision making. PMID- 11419227 TI - Time perspective and early-onset substance use: a model based on stress-coping theory. AB - This research tested the relation of time perspective to early-onset substance use (tobacco, alcohol, and marijuana) with a sample of 454 elementary school students with a mean age of 11.8 years. An adaptation of the Zimbardo Time Perspective Inventory (P. G. Zimbardo & J. N. Boyd, 1999) was administered with measures derived from stress-coping theory. Independent effects showed future orientation inversely related to substance use and present orientation positively related to substance use. Structural modeling analysis indicated that the relation of time perspective measures to substance use was indirect, mediated through behavioral coping and anger coping. Proximal factors for substance use were negative affect, peer substance use, and resistance efficacy. Results are discussed with respect to epigenetic models and the role of executive functions in self-control ability. PMID- 11419228 TI - Motivation for smoking cessation among pregnant women. AB - The authors evaluated an expanded measure of intrinsic and extrinsic motivation for smoking cessation in a population-based sample of 897 pregnant smokers (500 current smokers and 397 recent quitters). The measure assessed motivation related to pregnancy and parenthood in addition to general intrinsic and extrinsic dimensions. Current smokers at baseline who quit smoking by 28 weeks of pregnancy (n = 102) had significantly higher baseline levels of pregnancy-related motivation than continuing smokers. Extrinsic and pregnancy motivation dropped between baseline and 28 weeks of pregnancy among continuing smokers. Higher levels of intrinsic relative to extrinsic motivation at baseline were associated with sustained abstinence during the first 2 months postpartum. Results suggest that both general and pregnancy-specific motivation are important for smoking cessation and relapse prevention during pregnancy. Interventions to enhance the salience of health benefits over and above those related to pregnancy and other intrinsic benefits of a greater sense of self-control could protect against postpartum relapse. PMID- 11419229 TI - Effects of nicotine deprivation and irritability on physical aggression in male smokers. AB - Thirty-five continuing male smokers were randomly assigned to either a deprived or a nondeprived condition. On the basis of self-reported trait irritability, participants were then assigned to 1 of 4 groups: deprived-high irritable, deprived-low irritable, nondeprived-high irritable, and nondeprived-low irritable. Aggression was measured with a modified version of the Taylor aggression paradigm. Self-reported smoking urge was assessed throughout the reaction time competition. Results indicated that the deprived-high-irritable group displayed the highest levels of aggressive behavior. Positive associations between urge ratings and aggression indexes were detected only in the deprived high-irritable group. Results support the existence of an interactive effect of nicotine deprivation and trait irritability on physical aggression and highlight the role of smoking urge as an important factor in the deprivation-aggression link. PMID- 11419230 TI - Correlates of expressed and received violence across relationship types among men and women substance abusers. AB - This study examined expressed and received violence among men and women in substance abuse treatment. Rates of past-year partner violence (PV) did not differ by gender, although men reported markedly higher rates of nonpartner violence (NPV). Compared with PV, NPV was associated with more demographic and background factors (e.g., childhood aggression and conduct problems, family history of violence). The most consistent correlates of violence across relationship types were age, minority status, drug-related consequences, psychiatric distress, and frequency of childhood aggression. Only a few gender specific correlates were identified; most notably, witnessing father-to-mother violence was related to received PV only for women. Identification of correlates of expressed and received violence in partner and nonpartner relationships is essential for the assessment and treatment of individuals in substance abuse treatment settings. PMID- 11419231 TI - Prediction of obligatory exercise by exercise-related imagery. AB - Obligatory exercise is a compulsive behavior pattern in which exercise dominates daily life at the expense of other activities and lack of exercise produces withdrawal symptoms. This study examined the hypothesis that obligatory exercise is motivated similarly to eating disorders (cf. S. P. Coen & B. M. Ogles, 1993) and would be predicted by appearance-related imagery. Obligatory exercise (J. K. Thompson & L. Pasman, 1991) and exercise imagery (H. A. Hausenblas, C. R. Hall, W. M. Rodgers, & K. J. Munroe, 1999) were assessed before and after a 10-week exercise program in 243 adult exercisers. Regression analyses revealed that imagery accounted for 20% of the variance in obligatory exercise. Appearance related imagery did not predict significantly obligatory exercise. Energy-related imagery was the strongest predictor. Obligatory exercise may not be as associated with appearance-related concerns as eating disorders, suggesting different motivational bases for these 2 behavioral patterns. PMID- 11419232 TI - Effects of the "near miss" and the "big win" on persistence at slot machine gambling. AB - This study was conducted to examine the effects of the "near miss," when a player almost wins, and the "big win," a large early monetary gain, on persistence at slot machine gambling. One hundred eighty male and female undergraduates played a 4-wheel computerized slot machine. A 2 x 3 randomized factorial design and an acquisition-extinction procedure were used. The dependent variable, persistence, was defined by the number of trials in the extinction phase. As hypothesized, the near miss had a statistically significant effect on the number of trials in extinction. An inverted U-shaped effect of the near miss showed that the 30% near miss condition led to the greatest persistence, as compared to 45% and 15% near miss conditions. There was no statistically significant effect of the big win. Both cognitive and behavioral theories are discussed as explanations for effects of the near miss. PMID- 11419233 TI - A comparison of the predictive validity of four sets of baseline ASI summary indices. AB - This study compared the long-term predictive validity of original and new baseline Addiction Severity Index summary scores in methadone patients. The indices included the original Interviewer Severity Ratings (ISRs) and the new Clinical Indices (CIs), which use both lifetime and recent problem information, and the original Composite Scores (CSs) and Evaluation Indices (EIs), based on recent problems only. Outcomes were medical hospitalization, employment, alcohol intoxication, drug hospitalization, and psychiatric hospitalization in Months 7 24 poststudy entry and criminal charges in Months 0-24. Hierarchical logistic regression analyses were used in which 1 index was entered first and the other in the 2nd step. The reverse order of entry was used in a 2nd analysis. A final analysis set compared the best predictor from each of the 2 prior analysis sets. The CIs were superior to the other indices in predicting 3 of 6 outcomes (psychiatric hospitalization, drug hospitalization, and criminal charges); the EI was the best predictor of alcohol intoxication, and the CS the best predictor of unemployment. PMID- 11419234 TI - Manual-guided cognitive-behavioral therapy training: a promising method for disseminating empirically supported substance abuse treatments to the practice community. AB - A gap exists between empirically supported substance abuse treatments and those used in community settings. This study examined the feasibility of training substance abuse counselors to deliver cognitive-behavioral treatment (CBT) using treatment manuals. Participants were 29 counselors. Counselors were randomly assigned to receive CBT training or to a control group. Counselor attitudes were assessed pre- and posttraining. In addition, CBT therapy sessions were videotaped and rated for adherence and skillfulness. CBT counselors reported high levels of satisfaction with the training, intention to use CBT interventions, and confidence in their ability to do so. Ratings indicated that 90% of counselors were judged as having attained at least adequate levels of CBT skillfulness. Findings demonstrate the feasibility of using psychotherapy technology tools as a means of disseminating science-based treatments to the substance abuse practice community. PMID- 11419235 TI - Constructive thinking in adolescents with substance use disorders. AB - This study examined the concurrent validity and clinical correlates of the Constructive Thinking Inventory (CTI), a measure of experiential coping, in 551 adolescents aged 14-18 years with and without Diagnostic and Statistical Manual of Mental Disorders (4th ed.; American Psychiatric Association, 1994) substance use disorders (SUDs). The CTI was correlated with depression, anxiety, and conduct problems. After controlling for demographics and comorbid lifetime psychiatric disorders, the CTI scales of Behavioral Coping and Categorical Thinking distinguished adolescents with and without SUDs. Implications of these findings for assessment and treatment are discussed. PMID- 11419236 TI - Multidimensional assessment of perceived treatment-entry pressures among substance abusers. AB - Motivational assessment instruments typically measure clients' attributions about their readiness to change problem behaviors. They do not indicate why a client may be motivated to change, or provide guidance on how to retain an unmotivated client in treatment. The authors interviewed 415 substance abuse clients about their reasons for entering treatment and scored their responses along the dimensions of (a) negative versus positive treatment-entry pressures, (b) internal versus external sources of those pressures, and (c) the life domain from which the pressures emanated. Exploratory cluster analysis yielded 5 types of clients characterized by different profiles of perceived treatment-entry pressures. Cluster membership was predictive of treatment outcomes, and the clusters differed by demographic variables. These data support the discriminative and predictive utility of performing a multidimensional assessment of pressures to enter treatment. PMID- 11419237 TI - [Epileptogenic potential of sevoflurane]. PMID- 11419238 TI - [Electroencephalographic effects of sevoflurane in pediatric anesthesia: a prospective study of 20 cases]. AB - OBJECTIVE: To study electroencephalographic (EEG) changes in children during induction of anaesthesia with 8% sevoflurane. PATIENTS: Twenty patients were consecutively included (ASA I-II; aged: 13-101 months). MATERIAL AND METHOD: Prospective study; approved by the Ethics Committee; written informed consent obtained from parents; anaesthesia induced with 8% sevoflurane in oxygen; no premedication was given; to collect the EEG data, non-invasive electodes were installed before induction; EEG was recorded continuously and stored on a computer for later analyses (descriptive analysis). RESULTS: Myoclonics movements were observed during induction of anaesthesia in two boys. Myoclonics movements stopped spontaneously without therapy. Epileptiform EEG activity (spikes and poly spikes, burst suppression) was observed. In the 18 others cases, during sevoflurane induction, EEG changed rapidly with an increase in the range of beta activity (> 13 Hz) (n = 15) and in slow (< 8 Hz) and delta activity (< 4 Hz). In 14 patients, at time of laryngoscopy, the EEGs showed monophasic slow data activity (sharp high-voltage slow waves). Periods of EEG isoelectrical (burst suppression) were seen without spikes in four cases. CONCLUSION: At 2 MAC, epileptiform EEG activity has been observed during sevoflurane anaesthesia. In a short group of patients, this raises the question of avoiding sevoflurane in patients who have a history of epilepsy. PMID- 11419239 TI - [Recovery of neuromuscular block after continuous infusion of cisatracurium in patients with renal dysfunction]. AB - OBJECTIVE: Study of the recovery of neuromuscular block after continuous infusion of cisatracurium in patients with renal dysfunction. STUDY DESIGN: Prospective case-control study. PATIENTS: Forty adult patients scheduled for urological surgery were assigned to two groups according to the creatinine clearance (CC) as a measure of the renal function: group IR (CC < 60 mL.min-1) or group NR (CC > or = 60 mL.min-1). METHODS: After premedication with hydroxyzine, anaesthesia was induced with propofol, sufentanil and cisatracurium (0.15 mg.kg-1), and maintained using isoflurane, sufentanil and a continuous infusion of cisatracurium (0.12 mg.kg-1.h-1) adjusted for maintained a post-tetanic count < or = 5. Neuromuscular transmission was monitored at the adductor pollicis using accelerography (TOF Gard). Onset and recovery times in both groups were compared using Student's t test. RESULTS: Infusion time and total dose of cisatracurium were comparable in both groups. Onset times were 3.9 +/- 0.8 min and 3.5 +/- 0.6 min in groups IR and NR respectively. After the infusion, the time to train-of four ratio of 0.8 were not different in both groups: 77 +/- 18 min (group IR) and 73 +/- 13 min (group NR). However, the spontaneous recovery intervals 25%-75% were delayed in group IR (20 +/- 9 min vs 14 +/- 5 min p < 0.05). CONCLUSION: There are minor differences in the pharmacodynamics of cisatracurium between patients with normal or impaired renal function. Nevertheless, a marked interindividual variability in the recovery parameters was observed in patients with renal dysfunction. PMID- 11419240 TI - [Critical analysis of animal models of acute pain. II]. AB - OBJECTIVE: To analyse models of acute pain in experimental animals. DATA SOURCES: References were obtained from computerised bibliographic data banks (Medline and others) and the authors' personal documents. DATA SYNTHESIS: The majority of tests permit only a measurement of threshold, whereas clinical pain is almost always prolonged. The relationships between tests of acute pain and motor activity are reviewed from a number of standpoints; in particular we consider the influence, which postural adjustments of the animal may exert on motor responses in the limbs and the significance of flexor and extensor reflexes. In analysing the problem of the sensitivity of tests, we raise the following questions: 1) what type(s) of fibres underlie the observed responses and might these be different depending on whether one is stimulating a healthy or an inflamed tissue; 2) what significance do measurements of "latency" have when a stimulus is increasing; 3) how valid are the methods of analysing the results? The influence of species and the genetic line used in tests and the specificity and predictivity of tests are considered. Finally, we review those factors, which may distort behavioural measurements in animals, notably--pharmacokinetics, interactions between heterotopic stimuli, environmental factors and related psychophysiological/psychological considerations (subjective "undesirable" phenomena, learning phenomena). We pay particular attention to related physiological functions (thermoregulation, vasomotricity, blood pressure). These considerations lead us to re-position nociception within a much larger homeostatic framework which in addition to pain, includes phenomena such as anxiety and vegetative functions. They also suggest that we should define an "effective stimulus" as one, which activates nociceptive nerve terminals after a physical stimulus, has passed through a "peripheral lens" which regulates its intensity for reasons, which are physical, albeit of biological origin. Finally they remind us that the "system of pain" forms part of a whole set of subsystems- sensory, motor, vegetative, emotional, motivational--which scientific method, being reductionist by nature, cannot study in its entirety. However one must consider results of nociceptive tests within this general context. CONCLUSION: It is only by taking the approach described in this review, that fundamental and clinical research can interact usefully. PMID- 11419241 TI - [The effects of peridural anesthesia on duration of labor and mode of delivery]. AB - OBJECTIVE: To determine the effect of epidural analgesia (EA) on the duration of labour and the mode of delivery. DATA SOURCES: A Medline computerised literature research was conducted from 1989 to 2000 including all the prospective studies comparing EA and systemic analgesia during labour. DATA SYNTHESIS: EA prolongs the first and second stages of labour and increases the rate of instrumental delivery, without neonatal side effects. EA is not responsible for dystocia and caesarean section rate is not increased by this mode of analgesia. The effect of combined spinal-epidural analgesia is comparable to the one of EA on the length of labour and the mode of delivery. Ropivacaine does not appear to be different from bupivacaine in that setting. The benefit of ambulation remains controversial since it does not reduce the incidence of instrumental delivery, compared to conventional EA with similarly diluted local anaesthetic solutions. CONCLUSIONS: Pain relief provided by EA combined to modification of obstetric management (amniotomy, oxytocin) does not impair the rate of caesarean section and dystocia. PMID- 11419242 TI - [Acute involuntary intoxication with gamma-hydroxybutyric acid]. AB - Dangerous misuse of anaesthetic drugs is illustrated in a case of gamma hydroxybutyrate (gamma-OH) poisoning. A 36-year-old patient, coming from USA ingested approximately 200 mL of a liquid. He presented a non reactive coma (Glasgow coma scale at 3), abnormal movements, hypotension and bradycardia. Outcome was favorable with symptomatic treatment including tracheal intubation and ventilation. The patient was discharged 24 hours later. This case was a typical gamma OH poisoning. Its consumption is current in the United States for body-builders. Poisoning was involuntary, due to a dilution mistake. Toxicological analysis assessed this diagnostic. Such poisoning is exceptional in France. PMID- 11419243 TI - [Untreatable ventricular hemorrhage revealing cerebral metastasis of malignant melanoma]. AB - High levels of tissue-like plasminogen activator in cerebrospinal fluid reported in a patient with an intracerebral haemorrhage associated to cerebral metastasis of malignant melanoma could support the hypothesis of cerebral haemorrhage induced by tPA tumoral synthesis. PMID- 11419244 TI - [Purulent meningitis and bacteremia from Bacteroides fragilis revealing a meningioma]. AB - We report the case of a 78-year-old-woman with a meningitis caused by Bacteroides fragilis revealing a malignant tumor. The epidemiologic, etiologic and therapeutic features of these meningitis are reviewed. PMID- 11419245 TI - [Post-traumatic carotid-cavernous fistula]. AB - The authors report three cases of carotid-cavernous fistula occurring after severe cranio facial trauma. The diagnosis has been suspected on the association of a pulsatil exophtalmia and systolo-diastolic murmur, and confirmed by arteriography. Embolization was successful in two patients, the third one died from neurologic complication before embolization. The carotid cavernous fistula is a rare but severe complication of a craniofacial trauma. The functional (blindness) and vital prognosis (subarachnoid and intracerebral haemorrage). Treatment by interventional neuroadiology has considerably improved the outcome. PMID- 11419246 TI - [Fatal septicemia from Staphylococcus aureus resistance to glycopeptides]. PMID- 11419247 TI - [And now we present droperidol!]. PMID- 11419248 TI - [Expert conference: methodology developed by the Societe francaise d'anesthesie et de reanimation for the elaboration of recommendations]. PMID- 11419249 TI - [Acute pancreatitis (short text)]. PMID- 11419250 TI - [Agitation on induction with sevoflurane in children. Some elements in the response to clinical questions about sevoflurane]. PMID- 11419251 TI - [Early systemic antibiotic treatment for severe acute pancreatitis]. PMID- 11419252 TI - Joint Commission president outlines top strategic priorities to aid QI efforts. AB - At the top of the list of strategic priorities at the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) is patient safety. But Dennis O'Leary, MD, president of JCAHO, says his organization faces numerous challenges beyond that, including the value of accreditation itself. According to O'Leary, the underlying issue is that only about 15% of the 150,000 health care entities currently eligible for accreditation are actually accredited. PMID- 11419253 TI - Joint Commission faces several key challenges. AB - Dennis O'Leary, MD, says the Joint Commission made significant strides last year in widening the scope of the organization's reach across health care settings and refining the tools it uses to measure the quality of care provided. But he quickly adds that those accomplishments pale in comparison to the challenges that lie ahead. PMID- 11419254 TI - Aerospace engineering model identifies risks. AB - Issuing a new set of safety standards with which health care institutions must comply is all well and good, but offering those institution's creative tools to aid that compliance is even more significant. PMID- 11419255 TI - ED scores turnaround in patient satisfaction. PMID- 11419256 TI - DIGMAs (drop-in group medical appointments): satisfaction Rx for doctors and patients. PMID- 11419257 TI - New study gauges scope of nursing shortage. AB - One in five nurses now working is seriously considering leaving the profession within the next five years because of the conditions nurses face, according to a nationwide survey released April 19 by the Federation of Nurses and Health Professionals. PMID- 11419258 TI - It's time for a revolution in patient safety culture. AB - When implementing patient safety improvement initiatives, winning the hearts as well as the minds of people in your facility is important. Unless you convince people of the importance of the improvement endeavor, they will simply give it lip service. PMID- 11419259 TI - Partial benefit, total collapse. Providers mourn loss of Medicare services that never stood a chance. PMID- 11419260 TI - Technology: let the consolidation begin. Market slowdown fuels push toward fewer players. PMID- 11419261 TI - Children's mental health benchmarking: progress and potential. PMID- 11419262 TI - A challenge for the field: the association between violence and mental illness. PMID- 11419263 TI - Keeping it in the family. Bowen theory de-emphasizes therapist relationship. Interview by Colleen Fitzpatrick. PMID- 11419264 TI - Treating dementia: do conventional neuroleptics still have a role? PMID- 11419265 TI - Believe it: HIPAA's here. Our obligation to become educated about the law. PMID- 11419266 TI - A treatment plan for HIPAA anxiety disorder. PMID- 11419267 TI - Trusted networks: the state of healthcare security. PMID- 11419268 TI - What the inspector general's new compliance guidelines mean for you. PMID- 11419269 TI - HIPAA is here and here's what it means to your practice. PMID- 11419270 TI - HIPAA privacy regulations: what they mean for both physicians and patients. PMID- 11419271 TI - In Washington. Deadlock--or bipartisanship? PMID- 11419272 TI - After hard-fought campaign, Debbie Stabenow is literally in the hot seat. Interview by Ralph D. Ward. PMID- 11419273 TI - What a mouse can do for medical practice. PMID- 11419274 TI - Relationship-centered care: what do the Fetzer Institute and physician well-being have to do with it? PMID- 11419275 TI - Fetzer Institute proves to be a perfect fit for Thomas S. Inui, MD. PMID- 11419277 TI - MSMS and the alliance work to keep violent videos out of youths' hands. PMID- 11419276 TI - How nutrition found a home in our medical schools. PMID- 11419278 TI - Juvenile arthritis. More common than you think. PMID- 11419279 TI - Constitutionality of malpractice reform legislation. PMID- 11419280 TI - [Nobel prize-winning surgeons--part 2]. PMID- 11419281 TI - [Retinal detachment in the pseudophakic eye]. AB - Retinal detachments in pseudophakic eyes and their diagnostic and therapeutic features are analyzed. The authors pointed out diagnostic problems in detection of changes on the fundus significant for the onset of retinal detachment and difficulties in the detection of retinal rupture as important elements for the successful outcome of surgical treatment. The most frequent intra and postoperative risk factors for the occurrence of retinal detachment in pseudophakic eyes are presented. We have analyzed 16 patients treated by conventional surgical procedures as well as by inner silicone oil tamponade following pars plana vitrectomy in the Clinic of Ophthalmology of the Military Medical Academy during 1999. Results of surgical treatment of retinal detachments in pseudophakic eyes are in correlation with treatment results presented by other authors. PMID- 11419282 TI - [Risk factors for the onset of vasospasm and rebleeding after spontaneous subarachnoid hemorrhage]. AB - Retrospective study comprised 120 patients with spontaneous subarachnoid hemorrhage (SAH) and risk factors for vasospasm and rebleeding were analyzed. Rebleeding and vasospasm had the same incidence (26.7%). Vasospasm prolonged preoperative and postoperative hospital period (p > or = 0.05) and deteriorated the final treatment outcome (incidence of favorable outcome was decreased from 56.9% to 41.7%). Total rebleeding mortality rate was 62.5% and depended on the clinical state before rebleeding (p < 0.05). Risk factors for vasospasm were: poor clinical condition of a patient after the initial SAH, the period of 5-13 days after hemorrhage (p < 0.01) and aneurysm located on the anterior communicant artery (p < 0.01). Risk factors for rebleeding were: poor clinical condition of the patient after the initial SAH, the first 36 hours and the period of 8-11 days after the initial SAH, advanced age of the patient, severe arterial hypertension, vasospasm and aneurysm on the internal carotid artery. PMID- 11419283 TI - [Postoperative changes in left ventricular systolic function in patients with aortic valve stenosis]. AB - Left ventricular systolic function in aortic stenosis remains intact for long period due to myocardial hypertrophy. Aim of this study was postoperative follow up of the left ventricular systolic function both in patients with preserved and with decreased systolic performance. The study comprised 78 patients with severe and moderate aortic valve stenosis, out of whom 42 patients underwent surgery. In the majority of patients (88.1%) systolic function was improved, both in patients with preoperatively preserved systolic performance, and those who were with decreased systolic performance before the surgery. Systolic function was not improved in only 4.8%, while 3 (7.1%) patients died. Systolic function was improved mainly in the patients in whom systolic function disturbance was caused by the increase of afterload, and less frequently in patients with the decrease in myocardial contractility. PMID- 11419284 TI - [Clinical characteristics of chronic post-traumatic spinal neurogenic pain syndrome]. AB - In order to define clinical features of chronic posttraumatic spinal neurogenic pain syndrome we have analyzed a group of 23 patients with posttraumatic neurogenic pain. Patients (21 males and 2 females) were aged from 22 to 59 years, averagely 35.8 years. Painful sensations had the character of mechanical-incisive pain in 7 (30%) patients, and thermal pain equivalents in 3 (13%) patients. In 13 (57%) patients, sensor structure of this painful state had the character of associated thermal and mechanical painful sensations. Typical posttraumatic chronic spinal syndrome in our group of patients had persistent thermal qualities, of fluctuating intensity, with periodical, intensive pain onsets of mechanical character. Level of conus and cauda equina is predilectional level of the lesion for the development of intensive pain syndrome. Posttraumatic pain syndrome can be classified as intermittent and steady pain in relation to the rhythm of its manifestation, and as diffuse and dermatomal pain in relation to extension, in which painful state of occasional rhythm and dermatomal topography is the optimal indication for operative neurosurgical treatment. PMID- 11419285 TI - [Interleukin-1 beta in patients with primary immunocomplex glomerulonephritis]. AB - Interleukin-1 is one of the most important pro-inflammatory cytokines whose role in the pathogenesis of glomerulonephritic process was proved in numerous studies. The aim of this study was to determine the urinary level of this cytokine in patients with primary immunocomplex glomerulonephritis and its significance in diagnosis of this disease. This prospective study comprised a total of 96 patients (84 males and 12 females) with primary immunocomplex glomerulonephritis. The elevated urinary IL-1 beta level was noticed in 43 (49.4%) patients with different histological forms of glomerulonephritis. The mean concentration was significantly higher in patient's group (57.7 +/- 120.7 pg/mg creatinine) (range 1.1-731) compared to control group (10.2 +/- 5.96 pg/mg creatinine) (range 1.6 25.4) (p < 0.05). There was no significant difference in the frequency of elevated urinary IL-1 beta concentration in different patients group based on histological type of glomerulonephritis (chi 2 = 6.377, p > 0.05). On the basis of our results we concluded that the elevated concentration of IL-1 beta in majority of patients with primary immunocomplex glomerulonephritis had suggested its role in the pathogenesis of glomerulonephritic process. The urinary level of IL-1 beta represents a novel, non-invasive parameter in the diagnosis of this disease, but its measurement is not useful in predicting the histological type of primary immunocomplex glomerulonephritis. The results of our study suggest the possibility that urinary IL-1 beta level reflects the activity of glomerulonephritic process and it could be useful in non-invasive monitoring of the disease progression. PMID- 11419286 TI - [The soldier-senior officer relationship]. AB - One of the main conditions for the successful adaptation to the military is the relationship between soldier and his commander. The aim of this study was to determine interpersonal relationship between soldier and his commander, evaluated by soldier on the one side and by commander on the other. The sample consisted of 144 adapted (group A) and 400 nonadapted (group NA) soldiers. The applied instruments were two questionnaires: Soldier's evaluation of a commander and Commander's perception of a soldier. Commanders most frequently noticed: emotional reactions of soldiers (anxious and depressive symptoms), then their relation to basic training and performance of soldier's duty and then their adjustment in the group (isolation, problems with sleeping and eating). It was concluded that soldiers with the problems in communication with authority before the military also had the problems with military commanders. PMID- 11419287 TI - [Intrathoracic goiter]. AB - AIM: A goiter, situated partly or on completely in the thoracic cavity is usually referred to as intrathoracic goiter (ITG). However, inadequate definition of intrathoracic malpositions of thyroid gland caused usage of different classifications. This produced different and, often unclear criteria for the grouping of intrathoracic thyroid malpositions and, consequently, very different results and difficulties in any attempt of parallel review or analysis. The aim of this research was to analyze the ITG that include goiters with more than 80% of tissue in the thoracic cavity and intrathoracic thyroid choristomas, and to point out the diagnostic problems and surgical dilemmas. METHODS: Source of the used data were the studies published in the last 50 years in English, French, Italian, German and Russian language. Data have been integrated and combined for the detailed analysis, while quantitative synthesis was achieved by statistical analysis. RESULTS: ITG was represented in 1% of operatively treated thyroid diseases. One third of patients were asymptomatic. Commonly observed symptoms were dyspnea, stridor and dysphagia, and there was no significant difference in appearance of these symptoms between the two groups of ITG. Chest roentgenography and CT scan could provide the maximal information with no need for expensive procedures. Accuracy of scintigraphy in revealing and discerning of ITG is 70%, averagely. In 20% of patients correct preoperative diagnosis was not established. Therapy of choice in the treatment of ITG was surgical, even in asymptomatic cases. Thoracic approach was used in 95% of cases and there was no significant difference between the two groups of ITG. Total thyroidectomy or total lobectomy was definitely obligatory with contralateral subtotal lobectomy. There was no significant difference in the frequency of operative complications between ITG and non-ITG, and between the two groups of ITG. CONCLUSIONS: ITG is rare, but might present a difficult diagnostic and complex surgical problem. Common clinical presentation and identical surgical approach in operative treatment justify the common review of the two groups of ITG. With timely and adequately performed surgical approach, in specialized institutions, frequency of complications in operative treatment of ITG was not higher if compared to operative treatment of non-ITG. PMID- 11419288 TI - [Inhibitors of platelet glycoprotein IIb-IIIa in cardiology]. PMID- 11419289 TI - [Neurologic complications of hepatitis C]. PMID- 11419290 TI - [Personality disorders--the Cloninger biosocial model]. PMID- 11419291 TI - [Aortocaval fistulas]. AB - Aortocaval fistulas most frequently occur as a consequence of ruptured abdominal aortic aneurysms due to the additional trauma, spontaneous or iatrogenic. The basic symptomatology is the consequence of local irritation of spinal nerves, and major hemodynamic disturbances. In this paper we have presented two cases of aortocaval fistulas, treated at the Military Medical Academy with presentation of their preoperative status, performed diagnostic procedures, applied surgical procedures, and postoperative course and outcome. PMID- 11419292 TI - [Primary liposarcoma of bone]. AB - A case of 75-year-old man with extremely rare primary liposarcoma of the bone was presented. Stains for lipid, Sudan III, Sudan IV, and Oil Red "O", demonstrated the presence of intracellular lipid in the lipoblasts. Similarly, the S-100 immunoreactivity and electron microscopic findings of tumor cells confirmed the diagnosis of liposarcoma. Histochemical stains for PAS, Alcian-blue, mucikarmin, Toluidin-blue and Coloidal Iron were negative. PMID- 11419293 TI - [Acute intermittent porphyria as a problem in differential diagnosis]. AB - In this study the authors have stressed the multidisciplinary approach regarding the etiology of psychiatric disorders. This approach provides not only the quantitative but also the qualitative increase within the essence of psychic life and psychiatric disorders pointing out the integrity of biological, psychological and social in a human being. Many somatic diseases are followed by the psychiatric and neurological symptoms. One of these diseases is the acute intermittent porphyria (AIP) that is manifested with a wide spectrum of clinical manifestations and it is called "the great imitator". The AIP attacks are characterized by abdominal pains, neurologic and psychiatric disorders. The most important is to diagnose the disease at the onset. If not, many severe complications may occur, even the lethal outcome is possible. This case report has shown the significance of the above mentioned statements. PMID- 11419294 TI - [Analgesics-induced chronic renal failure in patients on dialysis therapy in Hungary]. AB - In recent years elaboration of the diagnosis of analgesic-nephropathy (ANP) with the help of imaging techniques significantly increased the possibility of diagnosing the disease. Therefore, evaluating the prevalence of ANP has become more accurate in our country as well. The prevalence of ANP has been investigated in patients who have newly been taken into the dialysis program due to renal disease of unknown aetiology in 22 dialysis centers between December 1994 December 1997. The diagnosis of ANP has been based on revealing chronic drug abuse in the history and positive results of renal imaging (decrease in length of both kidneys combined with either bumpy contours and/or papillary calcification). Among 284 patients dialysed with unknown diagnosis 42 (14.8% of all cases) proved to have ANP. All patients except 2 took analgesic mixtures containing phenacetin/paracetamol, phenason derivatives, acetilsalysilic acid, caffeine and/or codeine. According to their investigations, ANP is a common disease resulting in end-stage renal failure in Hungary as well. PMID- 11419295 TI - [The "watermelon" stomach as a rare cause of upper gastrointestinal bleeding]. AB - The watermelon stomach is a rare subtype of the gastric vascular malformations of unknown origin. It can usually be observed with autoimmune diseases, but in can be associated with other conditions. It is significant, since it can cause chronic iron-deficiency anaemia or sometimes serious acute blood loss. The typical endoscopic picture is linear red streaking of the antrum with convergence at the pylorus with visible tortuous small vessels. Histological examination is frequently not diagnostic. It demonstrates specific features including dilated mucosal capillaries with focal thrombi, dilated submucosal venous plexus and fibromuscular hyperplasia of the lamina propria. Therapy mostly is endoscopic, but some medical possibilities are also known. In their paper they report two cases of watermelon stomach, the lesions were successfully treated with oestrogen progesterone compounds. PMID- 11419296 TI - [Emergency treatment of severely ill newborns]. PMID- 11419298 TI - [Adrenomedullin under physiologic and pathologic conditions]. AB - Adrenomedullin (AM) is a novel 52-aminoacid-peptide hormone, originally isolated from human phaeocromocytoma. Adrenomedullin acts as a local autocrine and/or paracrine vasoactive hormone and has vasodilator and blood lowering properties, but its exact role is still uncertain. Adrenomedullin is considered to play an important endocrine role in various tissues maintaining the electrolyte and fluid homeostasis. Its normal plasma concentration is low. In hypertension, chronic renal failure and congestive heart failure its plasma concentration increases parallel to the seriousness of the disease. It is assumed that this peptide may be important under pathologic conditions compensating the effects of the vasoconstrictor molecules. Till now, investigations have proved that in diabetic angiopathies the levels and the production of vasoconstrictor factors and adrenomedullin were increased, while, those of other relaxing substances including nitrogenoxid were decreased. It is still uncertain whether increased release of adrenomedullin in diabetes is a compensatory mechanism or a coincidental event. Although, the precise role of adrenomedullin in the pathogenesis of diabetic complications is still to be elucidated, the elevated concentration of adrenomedullin in diabetes--which influences the vascular functions--let us speculate that there might be a certain interaction between adrenomedullin induction and vascular functions in diabetes. Thus, the induction of vascular adrenomedullin could be a new target of a therapeutic approach to the diabetic complications. PMID- 11419299 TI - [Treatment of acute necrotizing pancreatitis: a comparative study]. AB - The authors compared the therapeutic results of acute necrotizing pancreatitis during two separate time periods involving 97 and 24 patients, respectively. In the group "A", the time period lasted between May 31, 1991 and December 31, 1997. This group consisted of 97 patients, who, in addition to undergoing intensive treatment and antibiotic prophylaxis, later underwent late necrectomy and closed peripancreatic drainage. Jejunal nutrition only occurred in a limited number of 52 patients (53.6%), and was mostly used after a second operation. The group "B" consisted of patients from the time period between January 1, 1999 and December 31, 1999, and consisted of 24 patients treated in each case with intensive treatment in addition to antibiotic prophylaxis, early nasojejunal nutrition, and if necessary CT-guided percutaneous peripancreatic drainage was used. In the case of surgical intervention necrectomy and closed peripancreatic rinsing drainage was used. In group "A", significantly more early operations were performed than in group "B" (31 patients, 32.0% vs. 2 patients, 8.3%). In group "A" the number of reoperations was also higher as a total of 91 (83.5%) were reoperated as compared to group "B" in which a total of 18 (75.0%) reoperations took place. In the group "B", mortality was also lower (12.5%), than in the "A" group (17.5%). The authors attribute the successful therapy to the early jejunal nutrition, CT guided percutaneous peripancreatic drainage and to the introduction of prophylactic antibiotics. PMID- 11419300 TI - [Confirmation of a prenatal diagnosis of trisomy 13 with comparative genomic hybridization (CGH)]. AB - Trisomy 13 was diagnosed with genetic amniocentesis in a fetus of a 50 years old patient. Fetopathologic examination has shown cyclopy, proboscis and semilobar holoprosencephaly of the fetus, which is consistent with Patau syndrome. DNA was extracted from frozen liver tissue. Result of comparative genomic hybridization (CGH) was consistent with trisomy 13. They processed the DNA according Kallioniemi's method with modifications. CGH was developed for cancer genetics in mid 90s and now it is widely used in prenatal diagnosis too. CGH allows global analysis to detect unbalanced chromosome gains and losses in the whole genome in a single experiment without the need for cell culture. Significant results can be expected in those cases where conventional cytogenetics is not able to provide an answer either because postmortem tissue is not appropriate for cytogenetics or because the chromosomal change is sub-microscopical. CGH is a fluorescent in situ hybridization on a healthy target metaphase, with equal amount of competitive hybridization of green labelled digested test DNA and red labelled digested control DNA. Red to green ratio is assessed with the help of an image analyser. Green dominance represents chromosome gain, while red shift chromosome loss. In the paper they present the fetopathologic report of a trisomy 13 fetus and illustrate the method being the first Hungarian obstetric case diagnosed by CGH. PMID- 11419301 TI - From silence to voice. PMID- 11419302 TI - Also speaking from the periphery. PMID- 11419303 TI - On the power of intuition. PMID- 11419304 TI - Ethical issues in HIV research in poor countries. AB - PURPOSE: To increase awareness of the potential for conducting unethical research in poor nations in the name of scientific inquiry, and to enumerate ethical questions in HIV/AIDS research. ORGANIZING CONSTRUCT: Principles of ethics and ethical analysis in clinical research. SOURCES AND METHODS: Review of literature on ethical principles of research in developing nations. FINDINGS: People who participate in clinical trials in poor countries often do so because that may be the only way to gain access to health care. However, access to tested drugs beyond clinical trials is not guaranteed. Participants often do not understand the consequences of being research subjects. CONCLUSIONS: Conducting research in poor nations requires recognition of ethical issues and maintenance of ethical standards, regardless of material wealth of the countries. Ethical standards also indicate including people from the target population in decisions relating to designing and conducting clinical trials. PMID- 11419305 TI - On nursing theories and evidence. AB - PURPOSE: To expand the understanding of what constitutes evidence for theory guided, evidence-based nursing practice from a narrow focus on empirics to a more comprehensive focus on diverse patterns of knowing. ORGANIZING CONSTRUCT: Carper's four fundamental patterns of knowing in nursing--empirical, ethical, personal, and aesthetic--are required for nursing practice. A different mode of inquiry is required to develop knowledge about and evidence for each pattern. CONCLUSIONS: Theory, inquiry, and evidence are inextricably linked. Each pattern of knowing can be considered a type of theory, and the modes of inquiry appropriate to the generation and testing of each type of theory provide diverse sources of data for evidence-based nursing practice. Different kinds of nursing theories provide different lenses for critiquing and interpreting the different kinds of evidence essential for theory-guided, evidence-based holistic nursing practice. PMID- 11419306 TI - Misconceptions among nurses about evidence-based practice. AB - PURPOSE: To delineate the origins and fundamental tenets of evidence-based practice (EBP) and to enhance understanding of this important term. METHODS: A critical review of the literature pertaining to evidence-based practice from the fields of medicine and nursing, including international reports. FINDINGS: The roots of EBP indicate this term is not a synonym for research utilization but rather is the rubric for a set of epistemologic assumptions. Key components of EBP include evidence hierarchies and systematic reviews. Differences were found between the views of EBP presented in papers published by nurses within the US compared to those published by nurses outside the US. CONCLUSIONS: The literature shows incongruity in the interpretation of the basic tenets of the evidence-based paradigm. Additionally, nurses may underestimate implications of the evidence based movement. These two problems impede nurses' participation as full partners in the advancement of evidence-based practice. They also represent a call to action to achieve a more standardized framework for advancing EBP in nursing. PMID- 11419307 TI - Purpose in life and depressive symptoms in persons living with HIV disease. AB - PURPOSE: To examine relationships among purpose in life, HIV disease severity, demographic variables, and depressive symptoms in people living with HIV disease (PLWHIV). The hypothesis tested was that purpose in life is a stronger predictor of depressive symptoms than is HIV disease severity. DESIGN: Descriptive, correlational study using a convenience sample of 123 PLWHIV recruited from an urban infectious disease clinic in a university teaching hospital in the Southeast United States. METHODS: A self-administered questionnaire, including a sociodemographic tool, the Center for Epidemiologic Studies Depression Scale (CES D), and the Purpose in Life Scale (PIL), was used to collect data. Concurrent severity of HIV disease measures included HIV RNA viral load, CD4+ T-lymphocyte count, and the Revised HIV Medical Symptom Scale. Analytical methods included descriptive and inferential statistics and hierarchical regression analysis. FINDINGS: Depressive symptoms were greater and purpose in life was lower than in normative samples. Purpose in life was a stronger predictor of depressive symptoms than was HIV disease severity. CONCLUSIONS: Purpose in life was more important than were laboratory markers of disease progression for predicting depressive co-morbidity. Results from this study indicate the need for routine assessment of depressive symptoms in PLWHIV. Purpose in life should be explored as a potential buffer for depressive symptoms. PMID- 11419308 TI - African American women's experiences with breast cancer screening. AB - PURPOSE: To describe the experience and meaning of breast cancer screening for African American women. Breast cancer screening offers the greatest hope of reducing breast cancer mortality and improving breast cancer outcomes. Despite the proliferation of initiatives targeting African American women, they continue to be first diagnosed only when they have late-stage disease. DESIGN AND METHODS: Using hermeneutic phenomenological research methods, 23 low- and middle-income African American women were interviewed to gain an understanding of their experiences with breast cancer screening. FINDINGS: Participants varied in their experiences with breast cancer screening. Women spoke of a desire for a holistic approach to health that did not separate the breast from the rest of the body. This desire is indicated in the theme of minding the body, self, and spirit, along with themes of relationships and spreading the word about breast health issues. CONCLUSIONS: Interventions for African American women should include a focus on minding the body, self, and spirit to promote breast cancer screening, and should indicate the importance of relationships and spreading the word about breast cancer screening. PMID- 11419309 TI - A Quality of Health Outcomes Model for guiding obstetrical practice. AB - PURPOSE: To describe the application of the Quality of Health Outcomes Model, introduced by the American Academy of Nursing Expert Panel on Quality of Health Care, to obstetrical care, particularly second-stage labor. ORGANIZING FRAMEWORK: The model is different from the more linear structure-process-outcome frameworks used in the past to a dynamic conceptualization of reciprocal relationships among the system, intervention, client, and outcome components. Analysis of these components can provide a comprehensive picture of the complexity of patient care decision making in hospital labor and delivery units. CONCLUSIONS: Research findings indicate that interventions such as cesarean delivery and epidural analgesia may result in several significant quality of health outcomes for women that should receive increased attention. Uses of this model for shaping best practices among physicians and nurses and for setting directions for priorities in future interdisciplinary research and health policy are discussed. PMID- 11419310 TI - Are nurses adequately prepared for end-of-life care? AB - PURPOSE: To determine end-of-life (EOL) care core competencies and educational needs from practicing oncology nurses and to describe the characteristics of the respondents that are associated with selection of the leading core competencies. DESIGN AND METHODS: A researcher-developed mailed descriptive survey to members of the Oncology Nursing Society in Georgia, Virginia, Washington, and Wisconsin in late 1999. FINDINGS: Nearly all respondents indicated that EOL care was a part of their practice and that continuing education was important, but one-third of the respondents had less than 2 hours of continuing education in 2 years. How to talk to patients and families about dying was the top-rated core competency, consistent across age, educational level, practice role, and practice setting. Pain control and comfort care were also frequently selected as important EOL care issues about which more education is needed. CONCLUSIONS: Results show guidelines for improving educational curricula and considering characteristics of nurses when planning EOL educational programs. PMID- 11419311 TI - Ability of blind people to detect obstacles in unfamiliar environments. AB - PURPOSE: To describe obstacle detection, sometimes called face vision, and blind peoples' lived experiences. DESIGN: This Husserlian phenomenologic inquiry included a convenience sample of eight blind participants who possessed the ability to detect obstacles in unfamiliar environments. METHODS: Face-to-face conversations were audiotaped in a setting of the participant's choice. FINDINGS: Three themes were identified: "a sense that something is there," "sensing," and "space." These three entwined themes were recognized in the unity-of-meaning or uniting theme: sensing presence as open or closed space. CONCLUSIONS: The findings of this study indicate many questions for nurses. Given the synergistic recognition of obstacles that the participants described, secondary analysis of the data is indicated, for example regarding pattern recognition and human environmental field integrality. PMID- 11419312 TI - Transcultural adaptation of Champion's Health Belief Model Scales. AB - PURPOSE: To translate to the Arabic language, adapt, and test Champion's revised Health Belief Model Scales to measure Jordanian women's beliefs about breast cancer and breast self-examination (BSE). In Jordan, the primary site of cancer in women is the breast. No published studies have been found which describe women's beliefs or practices about breast cancer and BSE in Jordan. DESIGN: Descriptive correlational, using a cross-sectional survey with a random sample of 519 female university students and employees in Jordan, 1999 to 2000. METHODS: Champion's revised Health Belief Model Scales were translated to Arabic, validated by professional judges, back-translated to English, and pretested. Analyses included descriptive statistics of all the study variables, internal consistency, reliability estimates, construct validity using factor analysis, and predictive validity using multiple regression analyses. The dependent variables were the frequency of practice of BSE and the intention to practice BSE. FINDINGS: Factor analysis yielded nine factors: confidence 1, confidence 2, benefits, susceptibility, barriers, seriousness 1, seriousness 2, motivation 1, and motivation 2. All items on each factor were from the same construct. Significant correlations were found between the two confidence factors, the two motivation factors, and the two seriousness factors. Alpha coefficients ranged from .65 to .89. All the health belief variables accounted for 21% of the variance in the frequency of practice of BSE, and 7% of the variance in the intended frequency of practice. CONCLUSIONS: The translated version of Champion's scales was found to be a valid and reliable tool for use with Jordanian women. It can be used in planning and testing interventions to improve BSE beliefs and practice. PMID- 11419313 TI - Children's exposure to community violence. AB - PURPOSE: To analyze the effects of exposure to chronic community violence on children and adolescents. DESIGN: An integrative review of the literature was conducted on reports of studies about children's exposure to community violence. SOURCES: Studies for analysis were identified through a literature search of relevant topics in Medline, CINAHL, and PsycINFO. CONCLUSIONS: Exposure to community violence is related to significant stress and depression in children. Evidence on how exposure to violence affects children's growth patterns, intellectual growth, school performance, decision-making ability, or their hope for a future is needed. PMID- 11419314 TI - Youth access to tobacco in two communities. AB - PURPOSE: To describe the availability of tobacco to youth in two urban communities, as a basis for designing a community-based intervention program. DESIGN: Two Midwestern U.S. urban communities were the setting for unannounced tobacco compliance checks. The population of retailers for this study included all convenience stores, grocery stores, pharmacies, and discount stores licensed by the state to sell tobacco products in the two communities. Establishments where entry is restricted to adults were not included. The total population of retailers was used as the sample, and two purchases were attempted at each retail outlet. METHODS: Cigarette purchases by female teens and the location of cigarettes in stores were documented. Compliance-check procedures were planned with needed protections for retailers and youth. FINDINGS: Adolescent tobacco purchase rates were near the federal recommendations for the two urban communities. Retailer use of self-service displays was higher than expected. Availability of the designated brand for self-service was significantly associated with the purchase rate of tobacco by underage youth. CONCLUSIONS: Self service displays provide increased availability of tobacco to teens. Teen smoking and youth access to tobacco could be limited by legislation and policies to restrict retailer use of self-service displays. PMID- 11419315 TI - A response to California's mandated nursing ratios. AB - PURPOSE: To explore the need for evidence-based health policy, as illustrated by the mandatory staffing bill passed by the California state legislature in 1999. DESIGN: Prospective data were collected from a voluntary sample of California acute care hospital representatives to describe selected patient safety and clinical outcomes and nurse staffing variables at the patient-care unit level. METHODS: Data for descriptive analysis were collected on hospital nurse staffing, patient falls, and pressure ulcers from 257 medical, surgical, medical-surgical combined, step-down, 24-hour observation units, and critical care patient care units in 38 California acute care hospitals from June 1998 to June 1999. FINDINGS: Nursing staffing ratios varied among the 257 units. RNs provided 91% of the nursing care in critical care units. Patients in medical-surgical units received 59% of their care from RNs, 11% from licensed vocational nurses, and 30% from other caregivers. Preliminary data showed no relationships between reported staffing ratios in these hospitals and the incidence of patient falls or hospital acquired pressure ulcers. CONCLUSIONS: California Nursing Outcomes Coalition (CalNOC) data showed wide variations in staffing ratios, patient falls, and hospital-acquired pressure ulcers among nursing units and hospitals. These early findings indicate the need for additional research before determining minimal RN staffing requirements. Analysis of multiple sources of data may be necessary to determine safe staffing ratios and to provide evidence-based data for public policy. PMID- 11419316 TI - Benchmarking patient outcomes. AB - PURPOSE: To examine the usefulness of three types of benchmarking for interpreting patient outcome data. DESIGN: This study was part of a multiyear, multihospital longitudinal survey of 10 patient outcomes. The patient outcome used for this methodologic presentation was central line infections (CLI). The sample included eight hospitals in an integrated healthcare system, with a range in size from 144 to 861 beds. The unit of analysis for CLI was the number of line days, with the CLI rate defined as the number of infections per 1,000 patient line days per month. METHODS: Data on each outcome were collected at the unit level according to standardized protocols. Results were submitted via standardized electronic forms to a central data management center. Data for this presentation were analyzed using a Bayesian hierarchical Poisson model. Results are presented for each hospital and the system as a whole. FINDINGS: In comparison to published benchmarks, hospital performances were mixed with regard to CLI. Five of the 8 hospitals exceeded 2.2 infections per 1,000 patient-line days. When benchmarks were established for each hospital using 95% credible intervals, hospitals did reasonably well with only isolated months reaching or going beyond the benchmark limits. When the entire system was used to establish benchmarks with the 95% credible intervals, the hospitals that reached or exceeded the benchmark limits remained the same, but some hospitals had CLI rates more frequently in the upper 50% of the benchmarking limits. CONCLUSIONS: Benchmarking of quality indicators can be accomplished in a variety of ways as a means to quantify patient care and identify areas needing attention and improvement. Hospital-specific and system-wide benchmarks provide relevant feedback for improving performance at individual hospitals. PMID- 11419317 TI - Preparing for the future through genetics nursing education. AB - PURPOSE: To determine recommendations for curriculum change that are indicated by innovations in genetics. METHODS: Both quantitative and qualitative. The sample (n = 356) consisted of nurses identified as experts in genetics (n = 228) and nurses identified as potential users of genetics education (n = 128). Nurses' opinions of core components of a genetics curriculum were elicited via a mailed survey questionnaire. Participants also provided demographic information and completed the Jones Innovativeness Scale (1997). FINDINGS: Recommended content in genetics education for practicing nurses was identified by both groups of nurses. Innovativeness characterized 3% of the respondents. Ninety-eight percent of respondents said that adopting genetics education is important. In total, 398 items were identified as potential consequences of education that incorporates genetic information. CONCLUSIONS: Identified content provides a template for genetics education programs for nurses. Genetics nursing education was perceived to have positive outcomes for both nurses and clients. PMID- 11419318 TI - [Steps in development of social and hygienic monitoring]. AB - A new function--social and hygienic monitoring--is actively being put into activities of State Sanitary and Epidemiologic Service of Russian Federation. System of the monitoring is formed stepwise, with development of methodic, informational, material, technic and stuff resources in federal, regional and municipal establishments. Results of social and hygienic monitoring are used for better sanitary conditions, health care and environmental protection with consideration of regional features of sanitary and epidemiologic circumstances in territories of Russian Federation. PMID- 11419319 TI - [B-lymphocyte-related immunoglobulin biosynthesis in workers engaged in the yeasts hydrolysis production]. AB - 64 individuals having occupational chronic bronchitis and 41 healthy individuals- both working at yeast hydrolysis production--demonstrated increase in B immunity with higher spontaneous and antigen-stimulated Ig synthesis. The changes were dose-dependent. PMID- 11419320 TI - [Flow volume curve evaluation of respiratory function in wolfram-molybdenum miners]. AB - The authors studied respiratory function in workers of wolfram-molybdenum mine where dust contains nearly 75% of silicon dioxide and considerable amounts of metals. The examinees were those with I stage silicosis and with silicosis complicated by chronic obstructive bronchitis. The respiratory disorders appeared most severe in association of the diseases. PMID- 11419321 TI - [Microsomal monooxygenase activity after exposure to 1,4-dioxane]. AB - The author represents results obtained in evaluation of microsomal monooxygenases system in lungs, liver and kidneys of rats exposed to dioxane-1,4 fumes. Acute and chronic experiments proved activation of microsomal monooxygenases system in the subjected animals. PMID- 11419322 TI - [Evaluation of efficacy and quality of occupational health departments in the Centers of State Sanitary and Epidemiologic Control]. AB - The article deals with methodologic, methodic and organization features to evaluate efficacy and quality of activities performed by occupational hygiene specialists and departments in I authority level Centers of State Sanitary and Epidemiologic Supervision. The author suggests a method measuring quality of the activities and a computered technology for sanitary and epidemiologic evaluation. The technology could be used in control over Centers of State Sanitary and Epidemiologic Supervision in their activity aimed to preserve sanitary and epidemiologic well-being for industrial workers. PMID- 11419323 TI - [Aspects of silicotuberculosis course in workers of highly aluminous mullite refractories]. AB - The article covers differential diagnostic features of pulmonary tuberculosis and pneumoconiosis in workers engaged into mullite refractories production. The authors suggest that the disease courses as a new form of lung disorder- mullitosis. PMID- 11419324 TI - [Methodological basis for occupational monitoring in production of methylmethacrylate and polyurethanes]. AB - Passive dosimeters provide perfect personified evaluation of occupational intoxication danger for workers. Sanitary and chemical studies using methods designed by the authors were conducted in enterprises producing acrylic plastic, polyurethane, and medications. These methods enabled sanitary supervision over chemical hazards content of air, in coffee and work-wear. PMID- 11419325 TI - [Method to determine nitrite and nitrate ecologic load on humans]. AB - To evaluate ecologic load with nitrite-nitrate ions, the authors suggest determining the diurnal excretion and then excluding contribution of the endogenous compounds. PMID- 11419326 TI - [The investigation of cytogenetic chromosomal instability due to environmental hazards]. PMID- 11419327 TI - [Ambient air pollution in naturally doped ore processing]. PMID- 11419328 TI - [Influence of chemicals on female reproduction]. AB - Studying female reproductive health in city with developed chemical industry, the authors revealed significant difference in character and incidence of disorders between the chemical workers and general population. Calculated dependence of some reproductive disorders on values of chemicals in the air helps to forecast the risk degree. PMID- 11419329 TI - [Remote sequelae of combined influence of vinyl chloride and dichloroethane (experimental study)]. AB - Exposure to concentrations of vinyl chloride (VC) + dichloroethane (DCE) 50 + 50 mg/m3, VC + DCE 50 + 500 mg/m3 and VC + DCE 500 + 50 mg/m3 during pregnancy induced higher occurrence of fetal resorption, increased incidence of preimplantational and postimplantational fetal death. Carcinogenic activity of the same levels of VC and DCE was seen in various organs: high concentrations of DCE increased incidence of lung tumors, high levels of VC--of breast neoplasms, mesenterial lymphosarcoma and liver hemangiosarcoma. PMID- 11419330 TI - Assistance in the palm of your hand. Portable technology can free physicians from billing and coding chains. AB - PROBLEM: Paper-based billing system required unmanageable amount of paper, time and work. SOLUTION: Integration of handheld devices with ASP-based charge capture service. RESULTS: Increased accuracy in billing information, reduced incidence of denied claims, faster reimbursement, more accurate view of financial status. KEYS TO SUCCESS: The technology's ease of use, varied capabilities and early benefits. PMID- 11419332 TI - 2001 Healthcare Informatics 100. Profiles. PMID- 11419331 TI - The 2001 Healthcare Informatics 100. PMID- 11419333 TI - 2001 Healthcare Informatics 100. Up & comers. Breaking onto the scene. PMID- 11419334 TI - 2001 Healthcare Informatics 100. Mergers & acquisitions. Channeling resources. PMID- 11419335 TI - 2001 Healthcare Informatics 100. Partnerships. United we stand. PMID- 11419336 TI - 2001 Healthcare Informatics 100. Assessing ROI. Look before you leap. PMID- 11419337 TI - Healthcare innovations in technology systems. Telehealth in the city. The Charles R. Drew University-Nortel partnership delivers quality healthcare to an urban neighborhood. PMID- 11419338 TI - Analysis of expressed sequence tags from a cDNA library of somatic nuclear transfer-derived cloned bovine whole foetus. AB - The expression profile of genes in specific tissues is studied through analysing expressed sequence tags (ESTs) and provides useful information for characterizing gene function and tissue physiology. Analysis of ESTs is achieved by partial sequencing and characterization of clones isolated randomly from cDNA libraries. In the present study, we analysed the genes expressed in the somatic nuclear transfer-derived cloned bovine foetus in the early period of foetal development. To this aim, we constructed a directionally cloned cDNA library from somatic nuclear transfer-derived cloned 60 day-old whole foetus of cattle and sequenced 3' end of 510 randomly isolated clones. By BLASTN analysis, we identified 403 unique clones: 186 showed homology to previously identified genes, 123 matched uncharacterized ESTs and 94 showed no significant matches to sequences already present in DNA databases. Analysis of these cDNA clones revealed that this library contained a variety of functional genes, while foetuin, insulin-like growth factor 2, collagen type I alpha I and maternal G10 transcript genes were the most abundant transcripts. Our study allowed the establishment of a first list of genes expressed in bovine whole foetus. In future, the list of genes might help facilitate the understanding of physiology of foetal development in somatic nuclear transfer-derived cloned bovine foetus. PMID- 11419339 TI - Comparative mapping of chicken anchor loci orthologous to genes on human chromosomes 1, 4 and 9. AB - Comparative mapping of chicken and human genomes is described, primarily of regions corresponding to human chromosomes 1, 4 and 9. Segments of chicken orthologues of selected human genes were amplified from parental DNA of the East Lansing backcross reference mapping population, and the two parental alleles were sequenced. In about 80% of the genes tested, sequence polymorphism was identified between reference population parental DNAs. The polymorphism was used to design allele-specific primers with which to genotype the backcross panel and place genes on the chicken linkage map. Thirty-seven genes were mapped which confirmed the surprisingly high level of conserved synteny between orthologous chicken and human genes. In several cases the order of genes in conserved syntenic groups differs between the two genomes, suggesting that there may have been more frequent intrachromosomal inversions as compared with interchromosomal translocations during the separate evolution of avian and mammalian genomes. PMID- 11419340 TI - An allele associated with a non-detectable amount of alpha s2 casein in goat milk. AB - The goat CSN1S2 locus is characterized by the presence of three alleles, A, B and C, all associated with about 2.5 g/l of protein per allele. The SDS-PAGE analysis of 441 individual milk samples obtained from goats belonging to a population reared in Southern Italy showed that the milk produced by three goats did not apparently contain alpha s2-casein, whereas milk produced by 37 goats showed a less intense electrophoretic band of this casein fraction (about 50%). These results can be explained by hypothesizing the presence of another allele at this locus, CSN1S2o, associated with a 'null' content of alpha s2-casein. Southern blot, PCR and PCR-RFLP analyses of the DNA region containing the CSN1S2 gene of individuals producing milk with and without alpha s2-casein did not show differences between the two groups. As a consequence, goats producing milk without alpha s2-casein carry an apparently intact gene. The first results obtained by sequencing part of the CSN1S2o allele revealed a G-->A transition at nucleotide 80 of the 11th exon which creates a stop codon and could be responsible for the absence of the alpha s2-casein in goat milk. This mutation eliminates a NcoI restriction site. A test based on this polymorphism has been established in order to identify carriers of the CSN1S2o allele. PMID- 11419341 TI - Polymorphism of bovine major histocompatibility complex (MHC) class I genes revealed by polymerase chain reaction (PCR) and restriction enzyme analysis. AB - The polymorphic exon 2-exon 3 region of bovine major histocompatibility complex (MHC) class I genes was amplified by polymerase chain reaction (PCR) from genomic DNA samples with characterized class I polymorphism. The primers for amplification were designed in conserved regions at the borders of exons 2 and 3, based on all available cDNA sequences. The primers should, therefore, amplify most expressed class I genes, but may also amplify non-expressed class I genes. The PCR amplified class I gene fragments of 700 bp were characterized on the basis of restriction fragment length polymorphism (RFLP). The PCR-RFLP analysis of class I genes showed that the bands in each digestion could be classified as non-polymorphic, as shared between several bovine lymphocyte antigen (BoLA)-A types, or as specific to a single BoLA-A type. The same primers were then used for amplification of class I gene fragments from eight Sahiwal animals, a breed which originated in the Indian subcontinent. These studies showed that BoLA class I PCR-RFLP could be used to study class I polymorphism in family groups. PMID- 11419342 TI - The canine homeobox gene MSX2: sequence, chromosome assignment and genetic analysis in dogs of different breeds. AB - The MSX2 gene encodes a homeodomain transcription factor important for normal head and face morphogenesis. MSX2 is expressed in key craniofacial structures during development and mutations in the human gene give rise to various craniofacial abnormalities. We are interested in the genetic basis of non pathogenic variation in skull and face shape. As part of this study we have analysed DNA from a panel of different dog breeds, selected for the differences they show in these traits and investigated MSX2 as a candidate gene. In this paper we describe the cloning of the canine homologue of MSX2, the determination of its structure, sequence and localization of the gene to dog chromosome 4q23. The DNAs from 11 individual domestic dogs belonging to 10 different breeds were sequenced in a search for genetic variation. Our studies show that variation in MSX2 does not contribute to the diversity of face shape observed in these domestic dogs and that the MSX2 sequence is strongly conserved between different dog breeds. The proximal promoter shows a high level of interspecies sequence conservation and several conserved transcription factor binding motifs have been identified and their significance discussed. PMID- 11419343 TI - Physical anchorage and orientation of equine linkage groups by FISH mapping BAC clones containing microsatellite markers. AB - A horse bacterial artificial chromosome (BAC) library was screened for 19 microsatellite markers from unassigned or non-oriented linkage groups. Clones containing 11 (AHT20, EB2E8, HMS45, LEX005, LEX014, LEX023, LEX044, TKY111, UCDEQ425, UCDEQ464 and VIASH21) of these were found, which were from eight different linkage groups. The BAC clones were used as probes in dual colour FISH to identify their precise chromosomal origin. The microsatellite markers are located on nine different horse chromosomes, four of which (ECA6, ECA25, ECA27 and ECA28) had no previously in situ assigned markers. PMID- 11419344 TI - A microsatellite locus has more than one copy in the genome of two tilapia species (Oreochromis aureus and O. niloticus). PMID- 11419345 TI - Five new porcine genetic markers from a microsatellite enriched microdissected chromosome 13 library. PMID- 11419346 TI - The porcine ATPase, Ca++ transporting, plasma membrane 1 (ATP2B1) gene maps to chromosome band 5q23. PMID- 11419347 TI - An SNP is used to map MC1R to dog chromosome 5. PMID- 11419348 TI - Sequencing of four new BoLA-DRB3 and six new BoLA-DQB alleles. PMID- 11419349 TI - Polymorphic sites in exon 15 and 30 of the porcine C3 gene. PMID- 11419350 TI - Three polymorphic mitochondrial DNA markers for Perna canaliculus. PMID- 11419351 TI - Linkage assignment of two novel expressed sequence tagged sites in the chicken. PMID- 11419352 TI - Mapping of the luteinizing hormone/choriogonadotropin receptor gene (LHCGR) to chicken chromosome 3. PMID- 11419353 TI - Physical and linkage mapping of the canine phosphate carrier (SLC25A3) and apoptotic activating factor 1 (APAF1) genes to canine chromosome 15. PMID- 11419354 TI - Base substitutions in the sequences flanking microsatellite markers HMS3 and ASB2 interfere with parentage testing in the Lipizzan horse. PMID- 11419355 TI - Genetic variants detected by PCR-RFLP in intron 6 of the bovine calpastatin gene. PMID- 11419356 TI - Chicken microsatellite primers are not efficient markers for Japanese quail. AB - Domestic fowl or chicken (Gallus gallus) and Japanese quail (Coturnix japonica) belong to the family Phasianidae. The exchange of marker information between chicken and quail is an important step towards the construction of a high resolution comparative genetic map in Phasianidae, which includes several poultry species of agricultural importance. We tested chicken microsatellite markers to see if they would be suitable as genetic linkage markers in Japanese quail. Twenty-six per cent (31/120) of chicken primers amplified individual loci in Japanese quail and 65% (20/31) of the amplified loci were found to be polymorphic. Eleven of the polymorphic loci were excluded as uninformative because of the lack of amplification in some individuals or high frequency of nonspecific amplification. The sequence information of the remaining nine loci revealed six of them to contain microsatellites that were nearly identical with those of the orthologous regions in chicken. For these six loci, allele frequencies were estimated in 50 unrelated quails. Although the very few chicken markers that do work well in quail could be used as anchor points for a comparative mapping, most chicken markers are not useful for studies in quail. Therefore, more effort should be committed to developing quail-specific markers rather than attempting to adapt chicken markers for work in quail. PMID- 11419357 TI - Findings from a simulated disabled submarine survival trial. AB - Eleven volunteer submariners were exposed to simulated disabled submarine conditions for a maximum of 7 days to determine if the limited clothing and rations provided in escape compartments would compromise survival prospects. Daily rations were 0.568 liters of water (none on Day 1) and 100 g of barley sugar. The subjects wore working rig and the liner from the Mark 10 submarine escape and immersion equipment throughout, and slept in the outer dry suit. Air temperature fell from 22 degrees to 4.4 degrees C over 2 days and then remained at 4.4 degrees C. Although the subjects felt cold they were able to maintain their deep body temperature. The greatest threat to survival in this situation would be dehydration, one subject was withdrawn on Day 4 as his urine production over the previous 24 h was 130 ml and if not withdrawn and rehydrated this may have led to renal failure. Other medical problems suffered by the subjects during the 7 days included diarrhea, vomiting, hypoglycemia, headaches, and back pains, and, following the trial, non-freezing cold injuries to their feet. It is concluded that the rations are not adequate and could compromise the submariners ability to survive for 7 days in these conditions and during a subsequent escape procedure. PMID- 11419358 TI - Neutrophil sequestration and the effect of hyperbaric oxygen in a rat model of temporary middle cerebral artery occlusion. AB - A rat model of reversible occlusion of the middle cerebral artery was developed to assess the role of neutrophils and prophylactic hyperbaric oxygen (HBO2) on cerebral injury. Blood flow to the ipsilateral caudate putamen nucleus was reduced by approximately 50% during 2 h of arterial occlusion, but unaffected on the contralateral side. Neutrophil accumulation in brain was documented as myeloperoxidase concentration, which was elevated in both ipsilateral and contralateral cerebral hemispheres at 1 and 46 h after occlusion/reperfusion. HBO2 administered before ischemia at 2.8 atm abs for 45 min, as well as antibody induced neutropenia, reduced neutrophil accumulation, functional neurologic deficits, and cerebral infarct volume. These data demonstrate that one mechanism for benefit of HBO2 is related to its ability to ameliorate post-ischemic injury by inhibiting neutrophil sequestration. This mechanism should be taken into consideration when choosing partial pressures of oxygen for investigational clinical protocols. PMID- 11419359 TI - Ventriculo-peritoneal shunt performance under hyperbaric conditions. AB - A novice scuba diver with an implanted ventriculo-peritoneal (VP) shunt inquired about the performance characteristics of his shunt while diving. A literature search revealed no information regarding shunt performance under hyperbaric conditions. The manufacturer could not certify that the shunt would function under pressure. Therefore, four VP shunts were tested according to the manufacturer's testing protocol at 1 and 4 atm abs in a multiplace hyperbaric chamber. The pressure (in mm of H2O) required to establish flow through the shunts was recorded. Trials at 1 atm abs (n = 12) and 4 atm abs (n = 12) show that all shunts performed within the pressure range specified by the manufacturer. PMID- 11419360 TI - Adjunctive hyperbaric oxygenation in macular edema of vascular origin. AB - Macular edema (ME) is a primary reason for permanent decreases of visual acuity (VA) in diabetic retinopathy and retinal vein occlusions. The standard treatment (photocoagulation, rheological treatment) provide only a limited success. We have therefore studied the additional use of hyperbaric oxygenation (HBO2) in patients with persistent ME. Five patients (1 female, 4 male; 7 eyes) were treated by adjunctive HBO2. The average age of the patients was 60.6 (38.9-76.8) yr. The VA was measured with Early Treatment Diabetic Retinopathy Study charts before and after HBO2 with a monthly follow up. Fluorescein angiography was performed before and after HBO2 with a follow up every 3 mo. Each patient received 10-30 HBO2 treatments (median 15). The follow-up period was 15 mo. for every patient. The mean increase in VA was 3.5 (2-4) lines after HBO2. Retinal photocoagulation was performed in six eyes. Diabetic macular edemas showed no morphologic change, while ME originating from retinal vein occlusions (CME) regressed. The VA in our patients with ME of vascular origin seemed to improve with HBO2. Photocoagulation was necessary in most cases. Visual function correlated with the angiographic presentation only for CME. PMID- 11419361 TI - Effect of hyperbaric oxygen therapy on fetal spinal grafts: an experimental study. AB - In neural transplantation, lack of oxygen supply to the graft in acute stage is an important problem. This study was undertaken to evaluate the effects of hyperbaric oxygen (HBO2) on fetal spinal grafts. Spinal cord tissues obtained from 16-day fetal rats were transplanted into the spinal cord of adult Wistar rats (n = 30). After transplantation, they were randomly divided into two groups (n = 15 for each group); Group 1 received transplant alone, group 2 received transplant and HBO2 therapy consisting of 100% oxygen at 2.5 atm abs for 90 min twice a day for 7 days. Seven days after their surgery, all animals were killed for histologic examination. Degenerative changes, parenchymal integrity, host spinal cord edema, and vascularization of the graft were scored on a 3-point scale. Scores of these parameters were statistically analyzed. The graft survival rates in groups 1 and 2 were 54.5% and 71.4%, respectively. Animals treated with HBO2 showed statistically less spinal cord edema than the untreated groups (P < 0.05). Parenchymal integrity was also significantly better in this group (P < 0.05). The results indicate that edema reduction effect of HBO2 prevents the displacement of graft from the gap and contributes to the integration between the graft and host. PMID- 11419362 TI - Report on cardiovascular effect of hyperbaric oxygen in an acute anesthetized dog model. PMID- 11419363 TI - Guns in Wisconsin. PMID- 11419364 TI - CHILD SAFE. Working to prevent childhood injuries. PMID- 11419365 TI - Project UJIMA: working together to make things right. AB - OBJECTIVE: The purpose of this paper is to describe a group of pediatric patients who utilize services of Project UJIMA, a collaborative, community-based violence intervention/prevention program serving the greater Milwaukee metropolitan area. Program goals are to (1) reduce the rates of re-injury and premature death; and (2) minimize adverse psychological consequences of violence. METHODS: Retrospective observation of 218 patients who presented to an urban pediatric Emergency Department in 1998 as a result of interpersonal violent injury and received some level of service from Project UJIMA. RESULTS: Patient age ranged from 10 to 18 years with an average of 15 years. Sixty-nine percent (69%) of patients were male. The majority of patients were African-American (69%), followed by caucasian (21%), and Latino (8%). Physical assault was the leading type of injury (62%). Nearly one-third of cases were due to firearms (31%). Three youths required a repeat ED visit due to interpersonal injury. One hundred fifty six (72%) were referred for mental health services to address adverse psychological effects. CONCLUSION: Project UJIMA is approaching its goals of (1) reducing injury recidivism rates in this population; and (2) providing services to address related mental health issues. PMID- 11419366 TI - Firearm injury prevention: Internet resources for the health care provider. AB - This annotated bibliography examines public health and advocacy web sites related to firearm injury prevention. In 1998, on average, 1 child or adolescent died from firearm-related injuries almost every 2 hours, resulting in about 3800 youth deaths that year. Including adults, the death toll was 30,708. In this context, advocacy groups, consisting of health care professionals and community leaders, are reframing the concepts of firearm injury as a major public health issue. At the beginning of this new century, many of these advocacy groups are disseminating information over the Internet. Concurrently, professional medical organizations are encouraging health care professionals to stay updated on the latest health statistics, intervention techniques and legislative issues related to firearm injury prevention. In order to stay abreast of this rapidly changing information, physicians are utilizing Internet resources in a complimentary fashion to more traditional means of research (i.e. Medline). This article assists physicians in their online research of firearm injury prevention by providing a select list of effective, informative web sites. Despite the pitfalls of on-line searches, physicians with minimal time and effort can discover useful, complementary information on patient care, injury prevention and community advocacy. PMID- 11419367 TI - Trends in homicide in Wisconsin, 1985-1998. AB - OBJECTIVE: To explore homicide trends for Wisconsin over the period 1985-1998 and assess the state's progress towards meeting its year 2000 health objectives. METHODS: Wisconsin and US homicide data from the Centers for Disease Control's Web-based Injury Statistics Query and Reporting System (WISQARS) were analyzed for the period 1985-1998. RESULTS: Homicide rates in Wisconsin rose 54% between 1985 and 1995, but since that year, a sharp reverse in trends since has erased three-fourths of the increase. While Wisconsin rates for 1998 remain 15% above their 1985 levels, US rates peaked in 1991 and have since fallen to 12% below their 1985 levels. When compared to 1985, Wisconsin homicide rates for 1998 were 24% lower among whites, but 16% higher among blacks. CONCLUSIONS: Wisconsin failed to achieve its year 2000 objective for homicide reductions, with rates remaining over 80% above the state goal. Though Wisconsin's current homicide rate is nearly half the US average, recent reductions in Wisconsin homicide have lagged behind those observed nationally. PMID- 11419368 TI - Suicide trends in Wisconsin 1984-1998: good news for young and old. AB - PROBLEM: In Wisconsin, suicide is the eighth leading cause of death. Subgroups identified as being at higher risk are the elderly (> age 65) and young adults (aged 15-24). OBJECTIVES: 1) To compare overall national trends in suicide rates to Wisconsin trends in suicide rates from 1984 to 1998; 2) to examine suicide rates in Wisconsin sub-groups (age, gender and race) 1984-1998; and 3) to assess progress toward Wisconsin's Public Health Agenda for the Year 2000 suicide mortality goals. METHODS: National Center for Injury Control and Prevention data on suicide mortality were accessed for the United States and Wisconsin using WISQARS data extraction system. Mean suicide rates were calculated for two 5-year spans (1984-1988 and 1994-1998). Average numbers of deaths and percent change between mortality rates were calculated for the two 5-year spans. RESULTS: From 1984-1998, mean suicide mortality rates decreased 8% nationally and 14% in Wisconsin. Various trends are occurring between genders and ages. The current declines in teens 15-19 and elderly 75-84 may indicate reversals of previous trends. Rates for women decreased for nearly all age groups, continuing previously documented declines. While it appears progress has been made toward Wisconsin Public Health Agenda goals, it appears unlikely the overall objective of 8 per 100,000 will be met. PMID- 11419369 TI - Trends in falls, poisoning, drowning, and burns Wisconsin: 1986-1996. AB - Falls, poisoning, drowning, and burns comprise the four most common causes of unintentional injury death not related to motor vehicles. We examine mortality trends for these causes of injury in Wisconsin over a 10-year period (1986-1996). While national age-adjusted rates for fall mortality have remained relatively stable, Wisconsin has experienced a sharp 38% increase. The greatest increase in fall mortality was seen in the aged. Nationally, poisoning mortality rose by approximately 50% during this same period while Wisconsin saw almost no increase in mortality (3%) from poisoning. Wisconsin did experience an increase in deaths from poisoning in middle-aged adults. Mortality from drowning decreased by about 28% both in Wisconsin and in the United States, with much of the progress occurring in children and young adults. Burn mortality also declined nationally and within Wisconsin by 30% to 35%. The Wisconsin Public Health Department's "Healthier People in Wisconsin" objectives for the year 2000 will likely be met for drowning and burns, but not for falls. Overall, unintentional injury mortality in Wisconsin is decreasing. However, specific subpopulations are not reaping the benefits of this decline, suggesting a possible focus for future efforts aimed at lowering unintentional injury mortality. PMID- 11419370 TI - Buckling up: whose choice should it be? PMID- 11419371 TI - Recreational water safety in Wisconsin. AB - Due to Wisconsin's numerous lakes and rivers, tourists and residents alike frequently participate in water-related recreation. Unfortunately, drowning is a leading cause of injury-related death, especially in children and young adults. Developmental factors place children who are less than 5 years at increased risk for drowning. Teenagers also have a higher drowning mortality rate, largely due to risk-taking behaviors in this age group. This article reviews drowning mortality trends and statistics for Wisconsin and the United States, as well as current recommendations and legislation regarding water safety. We also discuss drowning prevention strategies such as patient education and legislative efforts surrounding swimming pool and boating safety. PMID- 11419372 TI - Child restraint systems: an update for physicians. AB - During the past decade, many advances have been made in motor vehicle safety restraint systems, and these advancements have brought changes in the recommendations for child passenger safety. In spite of these advances, a high morbidity and mortality rate continues to be attributed to improper or absent child restraint use. Child transportation safety is a complex issue. Lack of public awareness and incomplete laws contribute to the confusion. The solution involves professional and public education, further technological advances, and advocacy for improved legislation and regulation. Physicians have a unique opportunity to incorporate advice about proper restraint use into their daily practice; however, that requires a clear understanding of the current recommendations. Both the age and weight of the child determine the appropriate child restraint system. Understanding, practicing, and promoting these recommendations will save lives. This article reviews the current recommendations for proper child restraint while traveling in a motor vehicle. PMID- 11419373 TI - Falls in the geriatric patient. AB - Falls are common in the geriatric and older adult population, often causing significant morbidity or mortality. The geometry of the human body in motion requires a highly functional individual to remain balanced and upright under a variety of conditions and perturbations. Balance in this population is adversely affected by intrinsic and extrinsic factors. Medications' therapeutic and side effects frequently contribute to falls. An aggressive approach to falls reduction, including eliminating balance-altering medication, obtaining sub specialty and balance evaluations when warranted, and requesting home safety assessments is advocated in this review of the current concepts and literature. PMID- 11419374 TI - Geriatric trauma: review and recommendations. AB - Although the elderly make up about 12% of the US population, they account for nearly 33% of health care resources expended on trauma. A review of the literature in the area of geriatric trauma revealed a number of factors that influence the likelihood of injury in the elderly population. These include age related changes in postural stability, balance, motor strength, and coordination that may predispose individuals to falls and other accidents. It is important to have a high index of suspicion for injury and a low threshold for invasive monitoring and transfer to an intensive care setting. Considerations in the care of elderly trauma patients include comorbid conditions, pre-injury medications, reduced physiologic reserve, and the physical changes of aging. PMID- 11419375 TI - Wisconsin needs to support death investigation: here's why. AB - Medical examiners and coroners (ME/CO) can provide essential data for injury reporting and prevention, but often lack the resources, support and training to supply this important information. With increased interest in injury data, questions surrounding data collection and reporting are being raised. This article describes the experience of the Wisconsin Firearm Injury Reporting System, discusses results of a survey completed by Wisconsin ME/CO and offers recommendations for improved injury reporting and support for death investigation. PMID- 11419376 TI - Economic, population trends portend a physician shortage. PMID- 11419377 TI - Everyday life and legal values: a concept paper. AB - This "concept paper" emerged from a Law and Human Behavior (LHB) Workshop, that was called by the journal's Editor, Richard Wiener, and held at St. Louis University on March 19-21, 1999. This workshop, which brought together 22 scholars and researchers in legal psychology, was part of James Ogloff's Presidential Initiative Project for the American Psychology/Law Society, and was supported by St. Louis University and an NSF grant. Prior to our arrival, each participant answered queries from the Editor about LHB and the field of psychology and law, and each was asked to offer five topics that were underrepresented in the journal or that we would like to see addressed in future issues. At the workshop, we were assigned to small groups, and the authors of this paper constituted one such group. The "charge" for all groups was to "develop plans for encouraging submissions in areas of psycholegal scholarship that continue to be infrequent topics of investigation," and then to develop a concept paper. The direction our group took is captured by our title, "Everyday Life and Legal Values," and within this paper we explicate the topic, identify a number of underrepresented research areas, suggest some research paradigms for investigating them, and present this within a "perspectival directions" frame that ties established lines of research to the newer ones we propose. PMID- 11419378 TI - The impact of jury instructions on the fusion of liability and compensatory damages. AB - This study examined whether special jury instructions or the bifurcation of liability and compensation decisions would counter the tendency for evidence concerning the defendant's liability to affect damages awards. Mock jurors made liability and award decisions in response to a case description in which the level of defendant responsibility for the plaintiff's injuries and the type or timing of damages instructions were systematically varied. Instructions not to discount awards for uncertainty about the defendant's fault and instructions not to increase awards to punish the defendant's carelessness reduced the impact of the defendant's conduct on awards, while bifurcation did not. Additional findings suggest, at least in the context of the present study, that discounting may be a somewhat more potent process than surcharging. Possible explanations for these effects are discussed. PMID- 11419379 TI - Public eyewitness confidence ratings can differ from those held privately. AB - Despite much research on eyewitness confidence, we know very little about whether confidence ratings given in public might differ from those held privately. This study tested a prediction derived from self-presentation theory that eyewitnesses will give lower confidence ratings in public when there is a possibility of their account being contradicted by other witnesses as compared to when they report their confidence in private. In groups of 3 or 4 people, 96 participants watched a videotape of a simulated robbery and then answered 16 forced-choice questions about details from the videotape. In half of the experimental sessions, the participants shared their answers and confidence ratings aloud with the other participants (public condition), and in the other half, the answers and ratings were not shared (private). As predicted, confidence ratings were significantly lower in the public condition than in the private condition, but the privacy manipulation had no effect on response accuracy. These results are consistent with a self-presentation explanation, and they highlight the need to examine public confidence ratings more thoroughly. PMID- 11419380 TI - A preidentification questioning effect: serendipitously increasing correct rejections. AB - A study designed to test the effects of delay between crime and lineup on identification accuracy produced an unusually high rate of correct rejection from target-absent, simultaneous lineups (J. E. Dysart, 1999). Examining the procedures indicated that one question included in a preidentification questionnaire differed from those used previously. The question asked witnesses if they believed they would be able to correctly reject a target-absent lineup. An experiment (N = 138) was conducted to explore the impact of preidentification questions, including this new question, on witness accuracy. Results revealed that asking witnesses these questions, prior to viewing the lineup, significantly increased correct rejections of target-absent simultaneous lineups. PMID- 11419381 TI - On finding for defendants who plead insanity: the crucial impact of dispositional instructions and opportunity to deliberate. AB - One hundred twenty participants functioned as mock-jurors and as members of deliberating juries in an experiment designed to assess the impact of dispositional instruction on verdicts rendered in an insanity trial. Consistent with prior research (K. E. Whittemore & J. R. Ogloff, 1995), dispositional instruction had no effect on the verdict preferences of individual jurors prior to deliberating. Yet, as expected, the instruction manipulation had a major impact on postdeliberative decisions (i.e., group verdicts; individual juror verdict preferences). Content analyses of jury deliberations revealed that postdeliberative shifts toward harsh verdicts in uninstructed juries and toward lenient verdicts in instructed juries were mediated by the impact of the Instruction manipulation on the content of jury deliberations: uninstructed juries feared that an acquitted-insane defendant would be freed to act again, whereas instructed juries recognized that finding for an insane defendant implied his retention and treatment. Implications of these results for both legal policy and the conduct of mock-trial research are discussed. PMID- 11419382 TI - The effect of exposure to multiple lineups on face identification accuracy. AB - This study examines the conditions under which an intervening lineup affects identification accuracy on a subsequent lineup. One hundred and sixty adults observed a photograph of one target individual for 60 s. One week later, they viewed an intervening target-absent lineup and were asked to identify the target individual. Two days later, participants were shown one of three 6-person lineups that included a different photograph of the target face (present or absent), a foil face from the intervening lineup (present or absent), plus additional foil faces. The hit rate was higher when the foil face from the intervening lineup was absent from the test lineup and the false alarm rate was greater when the target face was absent from the test lineup. The results suggest that simply being exposed to an innocent suspect in an intervening lineup, whether that innocent suspect is identified by the witness or not, increases the probability of misidentifying the innocent suspect and decreases the probability of correctly identifying the true perpetrator in a subsequent test lineup. The implications of these findings both for police lineup procedures and for the interpretation of lineup results in the courtroom are discussed. PMID- 11419383 TI - Index of suspicion. Case 2. Diagnosis: Blunt hepatic injury. PMID- 11419384 TI - Index of suspicion. Case 3. Diagnosis: IBD (inflammatory bowel disease) and UC (ulcerative colitis). PMID- 11419385 TI - Defining energy for a new millennium. Proceedings of a symposium. Washington, DC, USA. April 4-5, 2000. PMID- 11419386 TI - Proceedings from Summit on Immune-Enhancing Enteral Therapy. May 25-26, 2000, San Diego, California, USA. PMID- 11419387 TI - Index of suspicion. Case 2. Diagnosis: mosaic Down syndrome. PMID- 11419388 TI - Index of suspicion. Case 3. Diagnosis: coarctation of the aorta (CoA). PMID- 11419389 TI - Index of suspicion. Case 4. Diagnosis: Toxocariasis. PMID- 11419390 TI - Velebit range--treasury of Croatian flora. PMID- 11419391 TI - Proceedings of the XVIII International Congress of the Transplantation Society. August 27-September 1, 2000. Rome, Italy. PMID- 11419392 TI - Hemostatic tools for the gastrointestinal surgeon: ultrasonic coagulator vs. bipolar ligation. PMID- 11419393 TI - Laparoscopic ultrasound for gastrointestinal surgeon. PMID- 11419394 TI - [A Pediatric case of lupoid leishmaniasis diagnosed by PCR]. AB - The purpose of this paper is to describe the clinical features of lupoid leishmaniasis in a child and to underline the use of PRC as a necessary and reliable tool in controversial diagnosis. Lupoid leishmaniasis, also known as chronic or relapsing leishmaniasis, is mainly widespread in the Middle East, where it represents up to 5% of all cutaneous Leishmaniasis. It strongly resembles Lupus Vulgaris, both clinically and histologically, and is therefore not usually diagnosed immediately but after a certain period of time. The amastigotic forms are rare or absent. The cutaneous nodules or plaques can slowly enlarge over the years. The case of an eleven-year-old albanian child living in Durazzo (Albania), suffering since three years with a plaque formed by apple jelly nodules and scars on his right cheek, is presented. Using PCR, the presence of Leishmania infantum DNA led to a diagnosis of lupoid leishmaniasis. The therapeutic strategy of a combination of oral itraconazole and infiltration of metilglucamina antimoniate has been carried out, with good result, as checked through "telemedicine". PMID- 11419395 TI - [Patient information, recommendations, guide for the physician. Text of recommendations. March 2000]. PMID- 11419396 TI - [Therapeutic management of patients with dyslipidemia. Text of recommendations. September 2000]. PMID- 11419397 TI - Scintimammography with 99mTc-sestamibi as a complementary diagnostic technique. PMID- 11419398 TI - [Correlation between protein-energy malnutrition and gingival status in children]. PMID- 11419399 TI - [The preprosthetic preparation of the endodontically treated abutment tooth. Post and core technique: a questionnaire analysis]. AB - The goal of the questionnaire was to determine the materials and methods that general dentists in Switzerland at present use for the treatment of non-vital abutment teeth. Questionnaires were sent to 1000 dentists from a computer generated random sample of the list of members of the SSO (Swiss Dental Society). The return ratio was 36%. The responses provided information on the current philosophies and prevailing techniques used to restore endodontically treated abutment teeth. Most of the answering dentists believed it was necessary to stabilize a root treated tooth with a post. The preferred restoration in anterior teeth was the cast post and core, while composite resin build-up with a prefabricated post was predominantly made for posterior teeth. Conical or combined cylindrical-conical posts, which had a sand-blasted surface, were predominantly used. Most of the answering dentists strove to stabilize the remaining tooth structure by circular enclosure of the tooth structure by the later crown (ferrule effect). Zinc phosphate cement was still preferred for cementation of metal posts and ceramic posts were most often fixed with the help of the acid etching technique. The most frequently described failures were retention loss or root fractures. PMID- 11419401 TI - How to be your own practice management information system consultant. PMID- 11419400 TI - [Decline of caries prevalence in Swiss military recruits between 1970 and 1996]. AB - In 1996, 416 military recruits (aged 20.4) were dentally examined. The mean DMFT based on clinical findings was 4,8 (including extracted premolars). The mean DMFT based on clinical and radiological findings was 5.3 (DT=l.2; MT=0.4: almost all of them premolars; FT=3.7. 13% of the recruits were caries-free (DMFT=0). On average, 8.7 affected sites (DFS) were counted; 1.4 were untreated (DS). The caries declined by 70% (DMFT) between 1970 and 1996 and by 46% (DMFT) and 54% (DFS) respectively between 1985 and 1996. In the latter period of time, the mean number of untreated lesions (DS) decreased by 71%, in approximal surfaces of molars and premolars even by 78%. The ratio FS/DFS was 51% in 1985 and 71% in 1996,. This means an improvement in restorative care. 56% of all DFS were found in pits and fissures, 36% in approximal surfaces of molars and premolars. 62% of all untreated lesions (DS) were observed in approximal surfaces of molars and premolars; this percentage was 83% in 1985. The estimated caries increment between age of 15 and 20 was approximately 1.6 DFS/year in the early eighties and 1.3 DFS/year in the early nineties. In 1996 , the former difference in caries prevalence between the recruits from the Canton of Vaud and those from other Western Cantons did not persist. This is probably due to an alignment of the nation-wide supply with fluoridated salt since 1983. PMID- 11419402 TI - Defining the profession. Pressure builds for patient safety. PMID- 11419403 TI - Water security for the 21st century--innovative approaches. Proceedings of the 10th Stockholm Water Symposium. Stockholm, Sweden, 14-17 August 2000. PMID- 11419404 TI - Validation of methods for determination of metabolic rate in the Edholm scale and ISO 8996. AB - The aim of this study was to validate the Edholm scale (Edholm, 1966) and the ISO 8996 standard (International Organization for Standardization [ISO], 1990) by comparing the metabolic rates estimated for both methods with the actual measured metabolic rate (MMeas) in 6 manual material handling tasks simulated under laboratory conditions. The metabolic rate was calculated from oxygen consumption VO2 (19 participants) according to Standard No. ISO 8996 (ISO, 1990). Additionally, the participants estimated perceived exertion using the Borg scale. The metabolic rates derived from the Edholm scale (MEdh) overestimated 5 of 6 activities by 34-50% (alpha = .05). The metabolic rates derived from ISO 8996 (MISO) overestimated all activities by 7-38% (alpha = .05). PMID- 11419405 TI - Acute disseminated encephalomyelitis associated with poliomyelitis vaccine. PMID- 11419406 TI - Application of flow cytometry to immunotoxicity testing: summary of a workshop. PMID- 11419407 TI - A scenic overlook on the road to clinical tolerance. PMID- 11419408 TI - Chicken and egg in autoimmunity and joint inflammation. PMID- 11419409 TI - Artherosclerosis as an infectious, inflammatory and autoimmune disease. PMID- 11419410 TI - The TGF-beta1 paradox in asthma. PMID- 11419411 TI - Autoimmunity mediated by innate immune effector cells. PMID- 11419412 TI - Early-embryonic culture and manipulation could affect genomic imprinting. PMID- 11419413 TI - ER confirmed as the location of mystery organelles in Arabidopsis plants expressing GFP! PMID- 11419415 TI - Summaries for patients. How willing are doctors to give patients with terminal cancer accurate information about their chances of survival? PMID- 11419414 TI - Summaries for patients. Associations between cancer and inflammation of the muscles. PMID- 11419416 TI - Summaries for patients. Eating fruits and vegetables helps prevent coronary heart disease. PMID- 11419417 TI - Summaries for patients. Two types of diets improve blood vessel function in men with high cholesterol. PMID- 11419418 TI - Summaries for patients. Impaired thinking in sick patients. PMID- 11419419 TI - Brainstem electronic implants for bilateral anacusis following surgical removal of cerebello pontine angle lesions. AB - The multichannel auditory brainstem implant (ABI) has been used successfully to treat deafness in individuals with neurofibromatosis type II. The device has been implanted in nearly 150 recipients worldwide, and clinical trials with the device are approaching completion. The implantation and fitting of the multichannel ABI differ significantly from cochlear implantation, and the processes are illustrated in a series of case studies. Performance data also are included from recipients with up to 7 years experience. PMID- 11419420 TI - From the Centers for Disease Control and Prevention. Trends in injection drug use among persons entering addiction treatment--New Jersey, 1992-1999. PMID- 11419422 TI - From the Centers for Disease Control and Prevention. Update: syringe exchange programs--United States, 1998. PMID- 11419421 TI - From the Centers for Disease Control and Prevention. Soft tissue infections among injection drug users--San Francisco, California, 1996-2000. PMID- 11419423 TI - JAMA patient page. Hip fractures. PMID- 11419424 TI - Comparison of early invasive and conservative strategies in patients with unstable coronary syndromes treated with the glycoprotein IIb/IIIa inhibitor tirofiban. AB - BACKGROUND: There is continued debate as to whether a routine, early invasive strategy is superior to a conservative strategy for the management of unstable angina and myocardial infarction without ST-segment elevation. METHODS: We enrolled 2220 patients with unstable angina and myocardial infarction without ST segment elevation who had electrocardiographic evidence of changes in the ST segment or T wave, elevated levels of cardiac markers, a history of coronary artery disease, or all three findings. All patients were treated with aspirin, heparin, and the glycoprotein IIb/IIIa inhibitor tirofiban. They were randomly assigned to an early invasive strategy, which included routine catheterization within 4 to 48 hours and revascularization as appropriate, or to a more conservative (selectively invasive) strategy, in which catheterization was performed only if the patient had objective evidence of recurrent ischemia or an abnormal stress test. The primary end point was a composite of death, nonfatal myocardial infarction, and rehospitalization for an acute coronary syndrome at six months. RESULTS: At six months, the rate of the primary end point was 15.9 percent with use of the early invasive strategy and 19.4 percent with use of the conservative strategy (odds ratio, 0.78; 95 percent confidence interval, 0.62 to 0.97; P=0.025). The rate of death or nonfatal myocardial infarction at six months was similarly reduced (7.3 percent vs. 9.5 percent; odds ratio, 0.74; 95 percent confidence interval, 0.54 to 1.00; P<0.05). CONCLUSIONS: In patients with unstable angina and myocardial infarction without ST-segment elevation who were treated with the glycoprotein IIb/IIIa inhibitor tirofiban, the use of an early invasive strategy significantly reduced the incidence of major cardiac events. These data support a policy involving broader use of the early inhibition of glycoprotein IIb/IIIa in combination with an early invasive strategy in such patients. PMID- 11419425 TI - Comparison of two platelet glycoprotein IIb/IIIa inhibitors, tirofiban and abciximab, for the prevention of ischemic events with percutaneous coronary revascularization. AB - BACKGROUND: In the setting of percutaneous coronary revascularization, agents in the class known as platelet glycoprotein IIb/IIIa inhibitors have significantly reduced the incidence of death or nonfatal myocardial infarction at 30 days. We assessed whether there are differences in safety or efficacy between two such inhibitors, tirofiban and abciximab. METHODS: Using a double-blind, double-dummy design at 149 hospitals in 18 countries, we randomly assigned patients to receive either tirofiban or abciximab before undergoing percutaneous coronary revascularization with the intent to perform stenting. The primary end point was a composite of death, nonfatal myocardial infarction, or urgent target-vessel revascularization at 30 days. The trial was designed and statistically powered to demonstrate the noninferiority of tirofiban as compared with abciximab. RESULTS: The primary end point occurred more frequently among the 2398 patients in the tirofiban group than among the 2411 patients in the abciximab group (7.6 percent vs. 6.0 percent; hazard ratio, 1.26; one-sided 95 percent confidence interval of 1.51, demonstrating lack of equivalence, and two-sided 95 percent confidence interval of 1.01 to 1.57, demonstrating the superiority of abciximab over tirofiban; P=0.038). The magnitude and the direction of the effect were similar for each component of the composite end point (hazard ratio for death, 1.21; hazard ratio for myocardial infarction, 1.27; and hazard ratio for urgent target vessel revascularization, 1.26), and the difference in the incidence of myocardial infarction between the tirofiban group and the abciximab group was significant (6.9 percent and 5.4 percent, respectively; P=0.04). The relative benefit of abciximab was consistent regardless of age, sex, the presence or absence of diabetes, or the presence or absence of pretreatment with clopidogrel. There were no significant differences in the rates of major bleeding complications or transfusions, but tirofiban was associated with a lower rate of minor bleeding episodes and thrombocytopenia. CONCLUSIONS: Although the trial was intended to assess the noninferiority of tirofiban as compared with abciximab, the findings demonstrated that tirofiban offered less protection from major ischemic events than did abciximab. PMID- 11419426 TI - Platelet glycoprotein IIb/IIIa inhibition with coronary stenting for acute myocardial infarction. AB - BACKGROUND: When administered in conjunction with primary coronary stenting for the treatment of acute myocardial infarction, a platelet glycoprotein IIb/IIIa inhibitor may provide additional clinical benefit, but data on this combination therapy are limited. METHODS: We randomly assigned 300 patients with acute myocardial infarction in a double-blind fashion either to abciximab plus stenting (149 patients) or placebo plus stenting (151 patients) before they underwent coronary angiography. Clinical outcomes were evaluated 30 days and 6 months after the procedure. The angiographic patency of the infarct-related vessel and the left ventricular ejection fraction were evaluated at 24 hours and 6 months. RESULTS: At 30 days, the primary end point--a composite of death, reinfarction, or urgent revascularization of the target vessel--had occurred in 6.0 percent of the patients in the abciximab group, as compared with 14.6 percent of those in the placebo group (P=0.01); at 6 months, the corresponding figures were 7.4 percent and 15.9 percent (P=0.02). The better clinical outcomes in the abciximab group were related to the greater frequency of grade 3 coronary flow (according to the classification of the Thrombolysis in Myocardial Infarction trial) in this group than in the placebo group before the procedure (16.8 percent vs. 5.4 percent, P=0.01), immediately afterward (95.1 percent vs. 86.7 percent, P=0.04), and six months afterward (94.3 percent vs. 82.8 percent, P=0.04). One major bleeding event occurred in the abciximab group (0.7 percent); none occurred in the placebo group. CONCLUSIONS: As compared with placebo, early administration of abciximab in patients with acute myocardial infarction improves coronary patency before stenting, the success rate of the stenting procedure, the rate of coronary patency at six months, left ventricular function, and clinical outcomes. PMID- 11419427 TI - Early gastric cancer in young, asymptomatic carriers of germ-line E-cadherin mutations. AB - BACKGROUND: Germ-line truncating mutations in the E-cadherin (CDH1) gene have been found in families with hereditary diffuse gastric cancer. These families are characterized by a highly penetrant susceptibility to diffuse gastric cancer with an autosomal dominant pattern of inheritance, predominantly in young persons. We describe genetic screening, surgical management, and pathological findings in young persons with truncating mutations in CDH1 from two unrelated families with hereditary diffuse gastric cancer. METHODS: Mutation-specific predictive genetic testing was performed by polymerase-chain-reaction amplification, followed by restriction-enzyme digestion and DNA sequencing in Family 1 and by heteroduplex analysis in Family 2. A total gastrectomy was performed prophylactically in five carriers of mutations who were between 22 and 40 years old. In each case, the entire mucosa of the stomach was extensively sampled for microscopical analysis. RESULTS: Superficial infiltrates of malignant signet-ring cells were identified in the surgical samples from all five persons who underwent gastrectomy. These early diffuse gastric cancers were multifocal in three of the five cases, and in one person infiltrates of malignant signet-ring cells were present in 65 of the 140 tissue blocks analyzed, representing in aggregate less than 2 percent of the gastric mucosa. CONCLUSIONS: We recommend genetic counseling and consideration of prophylactic gastrectomy in young, asymptomatic carriers of germ-line truncating CDH1 mutations who belong to families with highly penetrant hereditary diffuse gastric cancer. PMID- 11419428 TI - Images in clinical medicine. Left ventricular pseudoaneurysm. PMID- 11419429 TI - Deaths and injuries from house fires. AB - BACKGROUND: We sought to define the factors associated with house fires and related injuries by analyzing the data from population-based surveillance. METHODS: For 1991 through 1997, we linked the following data for Dallas: records from the fire department of all house fires (excluding fires in apartments and mobile homes), records of patients transported by ambulance, hospital admissions, and reports from the medical examiner of fatal injuries. RESULTS: There were 223 injuries (91 fatal and 132 nonfatal) from 7190 house fires, for a rate of 5.2 injured persons per 100,000 population per year. Rates of injury related to house fires were highest among blacks (relative risk, 2.8; 95 percent confidence interval, 2.1 to 3.6) and in people 65 years of age or older (relative risk, 2.6; 95 percent confidence interval, 1.9 to 3.5). Census tracts with low median incomes had the highest rates of injury related to house fires (relative risk as compared with census tracts with high median incomes, 8.1; 95 percent confidence interval, 2.5 to 32.0). The rate of injuries was higher for fires that began in bedrooms or living areas (relative risk, 3.7); that were started by heating equipment, smoking, or children playing with fire (relative risk, 2.6); or that occurred in houses built before 1980 (relative risk, 6.6). Injuries occurred more often in houses without functioning smoke detectors (relative risk, 1.5; 95 percent confidence interval, 1.0 to 2.4). The prevalence of functioning smoke detectors was lowest in houses in the census tracts with the lowest median incomes (P<0.001). CONCLUSIONS: Rates of injuries related to house fires are highest in elderly, minority, and low-income populations and in houses without functioning smoke detectors. Efforts to prevent injuries and deaths from house fires should target these populations. PMID- 11419430 TI - Respiratory syncytial virus and parainfluenza virus. PMID- 11419431 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 19-2001. A 50-year-old man with fever and joint pain. PMID- 11419432 TI - Future trials of antiplatelet agents in cardiac ischemia. PMID- 11419433 TI - Optimal treatment of acute coronary syndromes--an evolving strategy. PMID- 11419434 TI - The power of genetics to target surgical prevention. PMID- 11419435 TI - Vaccinating Japanese schoolchildren against influenza. PMID- 11419436 TI - Vaccinating Japanese schoolchildren against influenza. PMID- 11419437 TI - Vaccinating Japanese schoolchildren against influenza. PMID- 11419438 TI - Expenditures for the care of patients with HIV. PMID- 11419439 TI - Expenditures for the care of patients with HIV. PMID- 11419440 TI - Case 40-2000: gastric cancer in a young woman. PMID- 11419441 TI - Case 40-2000: gastric cancer in a young woman. PMID- 11419442 TI - Case 5-2001: unsuspected celiac disease. PMID- 11419443 TI - Deep venous thrombosis and thalidomide therapy for multiple myeloma. PMID- 11419444 TI - Management of the biliary tract in acute necrotizing pancreatitis. PMID- 11419445 TI - Endoscopic ultrasonography in acute biliary pancreatitis. PMID- 11419446 TI - Timing of and indications for biliary tract surgery in acute necrotizing pancreatitis. PMID- 11419447 TI - Noninvasive imaging of the biliary ducts. PMID- 11419448 TI - The clinical problem of biliary acute necrotizing pancreatitis: epidemiology, pathophysiology, and diagnosis of biliary necrotizing pancreatitis. PMID- 11419449 TI - Laparoscopic approach to the biliary tract in acute necrotizing pancreatitis. PMID- 11419450 TI - Early endoscopic management of acute gallstone pancreatitis--an evidence-based review. PMID- 11419451 TI - Toxicity and effects of adjuvant therapy in colon cancer: results of the German prospective, controlled randomized multicenter trial FOGT-1. AB - In this adjuvant three-arm multicenter trial, we studied whether modulating the standard 5-fluorouracil (5-FU) treatment with either folinic acid (FA) or interferon-alpha-2a (IFN-alpha) was superior to the recommended standard of adjuvant treatment in R0 resected colon cancer, 5-FU plus levamisole (LEV) for 12 months, in terms of toxicity and outcome. From July 1992 to October 1999, a total of 813 patients with resected colon cancer in stage II (T4N0M0; n = 63) or stage III (TxN1-3M0; n = 750) were randomized into three treatment groups and stratified according to N stage and participating centers (64 hospitals). The patients received a postoperative loading dose of 5-FU (450 mg/m2 on days 1 to 5 [arms A and C]) or 5-FU (450 mg/m2) plus FA (Rescuvolin, Medac, Hamburg, Germany, 200 mg/m2 on days 1 to 5 [arm B]). After completion of the first chemotherapy cycle, LEV was administered orally at a dosage of 150 mg per day on days 1 to 3, once every 2 weeks. After a 4-week chemotherapy-free interval, the treatment was continued weekly for 52 weeks. Treatment in one arm A ("standard") (n = 279) consisted of 5-FU intravenously (450 mg/m2 on day 1, once a week) plus LEV. 5-FU plus LEV was modulated in arm B (n = 283) with FA (200 mg/m2 on day 1, once a week) and in arm C (n = 251) with IFN-alpha at 6 million units three times a week repeated weekly. Treatment dosages were adjusted if toxic events above WHO grade 2 occurred. Patients were closely followed to determine recurrence and survival; the latter was calculated according to Kaplan-Meier analysis. Toxic events above WHO grade 2, mainly leukopenia, diarrhea, and nausea, occurred in 113 (14%) of 649 patients who had completed treatment in arms A (8.4%), B (13.5%), and C (31.7%). Discontinuance rates were as follows: 28% for all patients, 29% in arm A, 21% in arm B, and 34% in arm C. Overall relapse rates were 27% for all patients, 30% in arm A, 24% in arm B, and 28% in arm C. Relapses were local (8%), distant (78%), or combined (12%). Four-year overall survival rates in arms A, B, and C were 66.1%, 77.5%, and 66.2%, respectively. The 4-year survival rate in arm B was significantly higher compared to arm A (P <0.02, log-rank test) with arm A being equal to arm C. Adjuvant therapy with 5-FU plus FA plus LEV for 12 months is superior to the recommended standard (5-FU + LEV for 12 months). IFN-alpha modulation of 5-FU (plus LEV) adds to the toxicity with no therapeutic benefit. PMID- 11419452 TI - Behavior rating inventory of executive function. PMID- 11419453 TI - Statistical power and effect sizes of clinical neuropsychology research. AB - Cohen, in a now classic paper on statistical power, reviewed articles in the 1960 issue of one psychology journal and determined that the majority of studies had less than a 50-50 chance of detecting an effect that truly exists in the population, and thus of obtaining statistically significant results. Such low statistical power, Cohen concluded, was largely due to inadequate sample sizes. Subsequent reviews of research published in other experimental psychology journals found similar results. We provide a statistical power analysis of clinical neuropsychological research by reviewing a representative sample of 66 articles from the Journal of Clinical and Experimental Neuropsychology, the Journal of the International Neuropsychology Society, and Neuropsychology. The results show inadequate power, similar to that for experimental research, when Cohen's criterion for effect size is used. However, the results are encouraging in also showing that the field of clinical neuropsychology deals with larger effect sizes than are usually observed in experimental psychology and that the reviewed clinical neuropsychology research does have adequate power to detect these larger effect sizes. This review also reveals a prevailing failure to heed Cohen's recommendations that researchers should routinely report a priori power analyses, effect sizes and confidence intervals, and conduct fewer statistical tests. PMID- 11419454 TI - Patients, populations and policy: patient outcomes in chronic kidney disease. AB - Chronic kidney disease represents an interesting illustration for evaluating an epidemic of chronic illness, the impact of care processes and technology on health outcomes, the impact of financial incentives and cost containment on health outcomes, and the choices society must consider in responding to a chronic illness. The evidence suggests that strong economic pressures exist in the care of chronic kidney disease and that cost containment is important. The results in large part reflect the impact of economic pressures on clinical decision making in the absence of good evidence on outcomes. To improve clinical decision making we need valid evidence linking specific processes of care to patient outcomes. Specific processes amenable to study include the provision of preventive services, physician and nurse technical and interpersonal care and adherence to clinical practice guidelines. The ESRD Quality Study (EQUAL) currently underway and supported by the National Institutes of Diabetes and Digestive and Kidney Diseases, may help to guide physicians and centers in caring for their patients with chronic kidney disease. This investigation examines the relation between process of care and outcomes and expands outcomes measure to include disease specific quality-of-life measures and patient satisfaction and accounts for case mix using the Index of Co-Existent Disease, a measure of the extent of different comorbid diseases as well as their severity (18,19,20). Better data on how processes of care are linked to health outcomes can inform decision making and allow educated cost cutting and quality maintenance. PMID- 11419455 TI - [High altitude medicine]. PMID- 11419456 TI - CQ sources/bibliography. PMID- 11419457 TI - Density functional theory study of the fourier transform infrared and Raman spectra of dichloro-bis(2,4-pentanedionate)tin(IV). AB - Fourier transform infrared and Fourier transform Raman spectra of dichloro bis(2,4-pentanedionate)tin(IV) have been obtained. Density functional theory (DFT) BLYP calculations, have been carried out with the purpose of understanding the metal-ligand region spectra of this compound. Vibrational wavenumbers calculated by BLYP/6-31G* force fields are closed with the experimental results. The percentage of deviation of the bond lengths and bond angles gives a good picture of the normal modes, and serves as a basis for the assignment of the wavenumbers. The calculated geometrical parameters show slight differences compared with the experimental ones, and these differences can be explained by the different physical state of Sn(acac)2Cl2. The DFT-BLYP calculations assumed a free molecule in the gas phase. The experimental wavenumbers are obtained from the spectra of solid samples. PMID- 11419458 TI - Structural and models of dioxouranium(VI) with rhodanine azodyes--V. AB - The synthesis of several new coordination compounds of dioxouranium(VI) heterochelates with bidentate rhodanineazol compounds derived from rhodanine are described. The ligands and uranyl complexes have been charcaterized by various physico-chemical techniques. The bond lengths and the force constants have been calculated from asymmetric stretching frequency of O-U-O group. The infrared spectral studies showed a monobasic bidentate behaviour with the oxygen and azonitrogen donor system. The ligands contain intramolecular hydrogen bonds. PMID- 11419459 TI - The application of direct potential fitting to the X1sigma+ ground electronic states of LiCl, TlCl, RbF and CsF. AB - A procedure for directly fitting the potential energy curve of a diatomic molecule has been applied to the X1sigma+ ground states of LiCl, TlCl, RbF and CsF. Extensive, high-precision infrared and pure-rotational data sets for all isotopomers of the aforementioned molecules have been employed in direct least squares fits of a radially dependent Hamiltonian operator. The Born-Oppenheimer potentials are represented by a modified Lennard-Jones function that is shown to be flexible and provide the proper behavior in the long-range region of the potential. While the potential fits of LiCl and TlCl required the inclusion of Born-Oppenheimer breakdown functions, no such functions were necessary for either RbF or CsF. PMID- 11419460 TI - Local structure and local compressibilities around Co2+ impurity in ZnSiF6.6H2O crystal determined from electron paramagnetic resonance data. AB - The perturbation formulas of g-factors gparallel, gperpendicular and hyperfine structure constants Aparallel, Aperpendicular for 3d7 ion in trigonal octahedral crystal field are established on the basis of a cluster approach. These formulas consist of the contributions from configuration interaction and covalency effect and the parameters related to both effects can be estimated from the optical spectra and the structural data of the system under study. According to these formulas, the local trigonal distortion angle beta at pressure P = 0 and the local compressibilities d ln beta/dP in two pressure regions around Co2+ impurity in ZnSiF6.6H2O crystal are estimated by fitting the calculated electron paramagnetic resonance parameters gi, Ai and their pressure coefficients to the observed values. The results show that these local values are different from those of the host crystal because of the influence of impurity. PMID- 11419461 TI - Merocyanine-type dyes from barbituric acid derivatives. AB - The preparation and the solvatochromic behavior of two dyes, obtained by condensation of N,N'-dimethylbarbituric acid with dimethylaminobenzaldehyde and with 4,4'-bis(N,N-dimethylamino)benzophenone (Michler's ketone) are described. The latter dye is rather sensitive to the polarity of the medium, and in particular, to the hydrogen-bond-donor ability of protic solvents. The solvatochromism of both compounds is discussed in terms of the pi* and E(T)(30) solvent polarity scales and their differences in behavior interpreted with the aid of semiempirical calculations. PMID- 11419462 TI - Surface enhanced Raman spectroscopic studies of 1H-indazole on silver sols. AB - The SER spectra of 1H-indazole adsorbed on silver hydrosol were recorded in the 1800-100 cm(-1) and in the 3200-2800 cm(-1) regions. The SERS data were interpreted on the basis of previous vibrational assignments, with the help of the results of DFT calculations carried out using the 6-31G** basis. From the comparison of SER and normal Raman spectra it can be deduced that 1H-indazole is non-dissociatively adsorbed on metal surface and that it interacts with silver sol via nitrogen atoms and ring pi-system. The molecular plane assumes a tilted orientation with respect to the silver surface. The effect of varying the concentration of adsorbate was also evaluated. The observed changes of the relative intensities of some enhanced bands suggest that the molecule assumes a more tilted orientation upon lowering the concentration of the adsorbate. PMID- 11419463 TI - Vibrational studies of [Ni(II)(DIARS)2X]X, (DIARS=o-C6H4(As(CH3)2)2 and X=Cl, Br, I). AB - Infrared and laser Raman spectra of [Ni(II)(diars)2X]X, (X=Cl, Br and I) have been used as probes to determine the structures of chelated diarsine molecules. It has been observed that the effects of metal chelation and coordination geometry give rise to frequency shifts in these complexes. The variation in vibrational spectroscopic features indicates reduction in the symmetry of the complexes in the crystalline environment. The effect of halogen on the Ni-halogen stretching frequency of these square pyramidal complexes is not as significant as observed in the case of octahedral complexes. PMID- 11419464 TI - The mid-infrared (attenuated total reflection) spectroscopy of ethylene carbonate in water. AB - Ethylene carbonate (EC) and water solution compositions ranging from pure water to 60 mass% EC have been examined using infrared (attenuated total reflection) spectroscopy. The fundamental vibrational modes of EC in the mid-infrared between 2050 and 1000 cm(-1) were fitted to mixed Lorentzian-Gaussian bandshapes. The spectral data for EC bands between 1000 and 650 cm(-1) are also shown but were not curve-fitted due to baseline distortions from water librational modes. The results of the band analysis have provided information regarding the molecular structure of these solutions, and the fact that the structure is also concentration dependent. The Fermi resonance coupling between the v2 and 2v7 vibrations of EC have been analysed using a standard perturbation model. PMID- 11419465 TI - Application of FT-Raman spectroscopy to the characterisation of parchment and vellum, I; novel information for paleographic and historiated manuscript studies. AB - Raman spectra of parchment and vellum have been recorded and used to characterise materials used in mediaeval (10th century) and later works. These studies support our work on the characterisation of pigments used in historiated manuscripts and provide the basis for future studies of the interaction between pigments, binders and parchment or vellum. Examination of the undecorated parts of the manuscripts or books has been undertaken to ascertain if there are traces remaining of the procedures used to prepare the materials; in several cases, there is spectroscopic evidence for the presence of sulphates and slaked lime, which were used in the preparations of vellums for scriptoria. In one 14th century cantoral, there is evidence for paper being used as backing on the verso side of the vellum, probably from an early repair. PMID- 11419466 TI - Infrared and Raman spectra, ab initio calculations of structure and vibrational assignment of 1-fluoro-2-butyne. AB - The infrared spectra (3500-50 cm(-1)) of the gas and solid and the Raman spectra (3500-50 cm(-1)) of the liquid and solid have been recorded for 1-fluoro-2 butyne, CH3-C-triple bond-C-CH2F. Equilibrium geometries and energies have been determined by ab initio and hybrid DFT methods using a number of basis sets. A vibrational assignment is proposed based on the force constants, relative intensities, depolarization ratios from the ab initio and DFT calculations and on vibrational-rotational band contours obtained using the calculated equilibrium geometries. From calculated energies it is shown that the CH3 group exhibits almost completely free rotation which is in agreement with the observation of Coriolis sub-band structure in two of the degenerate methyl vibrations. The results are compared to the corresponding quantities for some similar molecules. PMID- 11419467 TI - Surface enhanced resonance Raman scattering imaging of Langmuir-Blodgett monolayers of bis (benzimidazo) thioperylene. AB - The synthesis, spectroscopic characterization and surface-enhanced spectroscopy of a new electro active organic material bis (benzimidazo) thioperylene (Monothio BZP) are reported. Langmuir monolayers of Monothio BZP were successfully formed on water subphase and characterized by the pi-A surface-pressure area isotherm. Langmuir-Blodgett (LB) monomolecular layers of Monothio BZP were fabricated onto glass substrates and onto silver island films for surface-enhanced spectroscopic studies. The results of surface-enhanced resonance Raman scattering (SERRS), SERRS imaging and surface-enhanced fluorescence (SEF) studies for Monothio BZP LB monolayers are reported. Raman imaging (global imaging and point-by-point mapping) of the SERRS signal for a single monomolecular layer on silver islands were obtained using the 514.5 nm laser line. The SERRS imaging permits a visualization of the variation of the SERRS intensity across of the rough metal surface. The SEF was recorded for the excimer emission of aggregates in the LB film. The distance dependence and the enhancement factor of SEF were determined using fatty acid spacing layers. A temperature dependence study of the LB monolayer SERRS and SEF spectra was carried out between -190 degrees and + 200 degrees C confirming the thermal stability of the LB monolayer on silver. The specificity and the sensitivity of SERRS signal on metal island films was probed using mixed LB films with 0.01% molecular ratio of Monothio BZP in Arachidic Acid (AA). The micro-Raman SERRS spectra from ca. 10(-3) attomole of the dye were recorded. PMID- 11419468 TI - Determination of photophysical rate constants for the non-protected fluid room temperature phosphorescence of several naphthalene derivatives. AB - The determination of kinetic parameters for luminescence processes is very important in understanding the phosphorescence process and the mechanisms of the heavy atom effect (HAE). In our previous work, we reported that room temperature phosphorescence (RTP) emission of many naphthalene derivatives can be induced directly from their aqueous solution without using any kind of protective medium, and the name Non-Protected Fluid Room Temperature Phosphorescence (NP-RTP) is suggested for this new type of RTP emission. In order to further understand this kind of luminescence phenomenon, the influence of heavy atom perturber (HAP) concentration on RTP lifetime of several naphthalene derivatives was studied in detail in this paper. The possibility of determination of photophysical parameters for emission of NP-RTP was explored based on the definition on the phosphorescence lifetime and the relation with the concentration of HAP in this paper. A static Stern-Volmer equation for phosphorescence was derived and the luminescence kinetic parameters were calculated. The results obtained by two different ways proved that photophysical parameters for RTP emission can be determined based on the changes of the RTP lifetime. PMID- 11419469 TI - Calculated gas-phase infrared spectra of 2,2,5,5,8,8-hexamethylhexahydroimidazo [1,2-a]-pyrazine-3,6-dione: relative performance of Hartree-Fock and hybrid density functional theory methods. AB - Geometric parameters and IR spectra of 2,2,5,5,8,8-hexamethylhexahydroimidazo[1,2 a]pyrazine-3,6-dione were computed by the HF, B3LYP, B3P86 and B3PW91 methods with the 6-31G(d) basis set. All the computation methods produce a boat conformation of the six-membered ring with the orientation opposite to the one experimentally observed, that is obviously due to incapability of such calculations to take into account mutual influence of the molecules in a crystal package. Quality of predicting the interatomic distances observed experimentally decreases in the series HF, B3LYP, B3PW91 and B3P86; for bond angles, in the series B3LYP, P3PW91, B3P86 and HF. At the same time, the IR spectra computed are in a good agreement with the observed FTIR spectrum. The correlation between the calculated and experimental vibration frequencies is characterized by the coefficients higher than 0.9999 for all three DFT methods; for HF it is slightly lower than 0.9995. The calculated absolute band intensities satisfactory match the observed relative intensities as well. Optimal uniform scaling factors calculated are 0.8973, 0.9593, 0.9540 and 0.9552 for HF, B3LYP, B3P86 and B3PW91, respectively. PMID- 11419470 TI - Spectral investigations on melanterite mineral from France. AB - EPR, Optical and IR spectral studies on a naturally occurring mineral melanterite are carried out at room temperature. EPR studies indicate the presence of Cu(II) ion in tetragonally distorted octahedral site and hyperfine lines could not be resolved due to higher concentration of the paramagnetic impurity in the mineral. Optical absorption spectrum is a characteristic of Fe(II) and Cu(II) ions. Crystal field parameters are evaluated. IR spectrum confirms the presence of water and sulphate ions. PMID- 11419471 TI - Experimental and theoretical vibrational study of bis(N,N dimethylthiocarbamoylthio)acetic acid. AB - Harmonic vibrational analysis at HF/3-21G level is performed on the ab initio optimized molecular structure of bis(N,N-dimethylthiocarbamoylthio)acetic acid, and the outcome used in assigning the infrared spectrum of the acid. The calculated spectrum compares well with the experimental solid-state FT IR spectrum recorded at 298 and at 77 K. PMID- 11419472 TI - Investigation of the thermal lens effect in water-ethanol mixtures: composition dependence of the refractive index gradient, the enhancement factor and the Soret effect. AB - The thermal lens effect produced in binary mixtures of water and ethanol has been investigated. It is shown that the sensitivity of the thermal lens method upon the addition of ethanol in water varies as the temperature-dependent refractive index gradient to thermal conductivity ratio of the mixture. The dependence of k and dn/dT upon the ethanol volume fraction follows a second-order and fourth order polynomial, respectively, and cannot be precisely predicted using the additive rule. Moreover, depending on the experimental conditions and the mixture composition, the temperature gradient produced subsequently to relaxation of the excited species induces mutual migration of the solvent molecules and the formation of a concentration gradient in the irradiated area. This effect, known as the Soret effect, can locally change the thermo-optical properties of the solution and produce an additional signal, especially when steady-state experiments are done. This may result in errors as large as 20% when quantitative informations have to be derived from experimental data. PMID- 11419473 TI - Effect of solvent on the cis-trans conformational equilibrium of a proline imide bond of short model peptides in solution. AB - Proton NMR spectra of proline-containing short peptides with N-terminal sequences of N-acetyl-prolyl- (Ac-Pro-) N-tert-butoxycarbonyl-phenylalanyl-prolyl- (Boc-Phe Pro-) and N-tert-butoxycarbonyl-leucyl-prolyl- (Boc-Leu-Pro-) were measured in mixed solvents of hexadeuterodimethylsulfoxide and deuterochloroform (CDCl3). Population ratios of cis and trans conformers with respect to the proline imide bond and chemical shifts of NH protons were obtained as a function of a CDCl3 fraction of solvent. With increasing fraction of CDCl3, the trans percentages of the Ac-Pro-imide bonds increased. On the other hand, those of Boc-Phe-Pro- decreased, and those of Boc-Leu-Pro- exhibited middle tendency. From the solvent dependent variation of the chemical shifts of the NH protons, intramolecular hydrogen bonds that stabilize the trans form of Ac-Pro- and the cis form of Boc Phe-Pro- were discussed. For the Ac-Pro- peptides, only the trans forms are found to the compatible with 7-, 10-, and 13-membered hydrogen-bonded rings that would be similar to the ordinary secondary structures, gamma- and beta-turns and alpha helix, respectively. For the cis form of Boc-Phe-Pro-R (R = O-methyl or glycyl-O ethyl), the hydrogen-bonded structure is found to be similar to the type-VIa beta turn. On the other hand, for Boc-Phe-Pro-Pro-Leu-Gly-NH2, it has been suggested that two different hydrogen bonds, which are different from that of the type-VIa beta-turn, support each other and cooperatively stabilize the cis form. PMID- 11419474 TI - Fluorescence characteristic study of the ternary complex of fluoroquinolone antibiotics and cobalt (II) with ATP. AB - The results from the measurement of the fluorescence spectra of fluoroquinolone antibiotics including ofloxacin (OF), norfloxacin (NOR) and ciprofloxacin (CIP) complexed with cobalt (II) and ATP give information concerning the antibiotics nucleotide interactions. From the fluorescence spectral data, it appears that the fluoroquinolone antibiotic cannot directly complex with ATP but indirectly complex with cobalt (II), which is playing an intermediary role. The interaction of fluoroquinolone antibiotic with the nucleotide occurs mainly through the phosphate group. The conclusion offers a more complete mechanism, which is important for understanding the interaction of these drugs with DNA. PMID- 11419475 TI - A study of amber and copal samples using FT-Raman spectroscopy. AB - FT-Raman spectra were collected from fossil resins originating from a variety of geographical locations. The spectral profiles of most of the fossil resins could be related to modern resins containing diterpenoid components with predominantly labdane skeletons. The spectra collected from the fossil resin sample from Borneo differed from other fossil resins and was found to contain triterpenoid components. The differences in the spectral profile of fossil resins containing diterpenoid components are shown to relate to differences in level of maturation rather than geographical origin. FT-Raman spectra of fossil resins cannot be used to distinguish source although the degree of maturation can be used as an indicator to narrow the range of possible geographical origins. PMID- 11419476 TI - A 72-year-old man with confusion and nonoliguric renal failure. PMID- 11419477 TI - Mitral valve prolapse: a review of the literature. PMID- 11419478 TI - Polarities of the centriolar structure: morphogenetic consequences. AB - Centrioles and basal bodies are two versions of the same conserved eukaryotic organelle and share two remarkable properties: nine-fold symmetry of their microtubular shaft and capacity to generate a new organelle in a fixed geometrical relationship to the mother organelle. It can thus be postulated that what is true for basal bodies is likely to be true also for centrioles. While the functions of centrioles are difficult to dissect, the functions of basal bodies are easier to approach. Over more than two decades, studies on protists have led to the notion that ciliary and flagellar basal bodies display polarities, not only a proximo-distal polarity, like in centrioles, but also a circumferential polarity accorded to the polarities of the cell and of its cytoskeleton. The major cytological and genetical data, mainly of Chlamydomonas, Paramecium and Tetrahymena, which support the notion that the microtubule triplets of basal bodies are non-equivalent, are reviewed. The morphogenetic implications of this circumferential anisotropy, perpetuated through the process of basal body duplication itself, are discussed. The question is raised of the possibility that centrioles also display a circumferential polarity, like basal bodies, and whether at least certain of their functions depend on such asymmetries. PMID- 11419479 TI - Photon beam dosimetry at a blocked beam edge using diffusion approximation. AB - A simple analytical model is presented for the transport of secondary electrons at a photon beam edge using the energy averaged solution of the Boltzmann equation, originally developed for beta-ray dosimetry at a plane interface. Dose at a point under a block is assumed to be due to secondary electrons and the scattered photons generated from the primary photon beam. The diffusion approximation is used for the secondary electron transport at a virtual plane interface created by the block. The dose from the scattered photon component is treated as decaying exponentially with distance from the beam edge. Comparisons made with the model and measurements are in general agreement for high energy accelerator beams. PMID- 11419480 TI - Single-molecule detection of DNA separations in microfabricated capillary electrophoresis chips employing focused molecular streams. AB - DNA separations have been performed using microfabricated capillary electrophoresis chips and detected using a single-molecule fluorescence burst counting technique. We enhanced the percentage of electromigrating DNA molecules that were detected by focusing the sample through the 1-microm-diameter focused laser beam. The sample was focused by introducing a taper in the separation channel and by a sheath flow delivered from cross channels. The sample stream width, single-molecule velocities, and single-molecule count rates varied linearly with the current density ratio as expected. The optimal laser power for each focusing condition was investigated using dilute solutions of pBluescript DNA. Although fluorescence burst heights and background varied with laser power, the signal-to-noise ratio was only weakly dependent on this parameter using our single-molecule counting technique. Focused single-molecule counting was used to detect separations of a 100-1000-bp DNA sizing ladder. The increase in molecular detection efficiency was quantified by applying a focusing current midway through the ladder separation and comparing observed count rates to the known molecular concentration in the bands. The molecular detection efficiency varied linearly with the applied current density ratio, and greater than 3-fold enhancements in detection efficiency were achieved. PMID- 11419481 TI - Barrier durability of latex and vinyl medical gloves in clinical settings. AB - This study evaluated changes in the tensile strength and barrier integrity of medical gloves during hospital clinical use. Nonsterile vinyl, sterile vinyl, and nonsterile natural rubber latex gloves were collected after use in a clinical setting and then tested for tensile strength, elongation, and water leakage. Tensile properties of vinyl gloves did not change during use, whereas changes in latex depended on the brand evaluated. New gloves, regardless of material of manufacture, were found to have leakage rates of 2% or less. Two brands of nonsterile vinyl gloves were found to have an average rate of leakage after use of 24 to 28% (average 26%), three brands of latex gloves of 6 to 10% (average 8%), and one brand of sterile vinyl gloves of 3%. Low-protein powderless latex gloves leaked slightly less than the powdered brands. The high rates of leakage observed for nonsterile vinyl gloves indicate that they provide less barrier protection than latex in typical hospital use. The low leakage rate observed for sterile vinyl gloves indicates that barrier durability is not solely a function of the generic polymer composition of the barrier. PMID- 11419482 TI - Faculty of Family Planning and Reproductive Health Care. Royal College of Obstetricians and Gynaecologists. Guidance April 2000. Emergency contraception: recommendations for clinical practice. PMID- 11419483 TI - The complete DNA sequence of the nuclear ribosomal RNA gene complex of Verticillium dahliae: intraspecific heterogeneity within the intergenic spacer region. AB - The complete sequence of the nuclear ribosomal DNA gene complex of the phytopathogenic fungus Verticillium dahliae has been determined. The tandemly repeated unit was 7216 bp long and appears to be the shortest rDNA cluster described so far among filamentous fungi. Primer pairs were designed for amplification of the region spanning half of the 28S subunit, the intergenic spacer (IGS), and the 5' end of 18S subunit of a number of Verticillium strains, isolated from various hosts and geographic origins. Great heterogeneity was detected in the amplified products of the IGS region resulting in fragments varying from 1.6 to 2.0 kb. The majority of Verticillium isolates were classified into two groups with 1.6- and 1.7-kb amplified products, respectively. The former group included 31 V. dahliae, 7 V. longisporum, and 1 V. albo-atrum isolates, whereas the latter included 10 V. dahliae and 1 V. albo-atrum isolates. Sequence analysis of representative PCR products of the above groups identified a "hot spot" region harboring most of larger insertions, whereas most of the small changes were due to transitions and transversions. One V. longisporum isolate with a 2.0-kb PCR product contained 13 perfectly conserved tandem repeats of 39 bp long. The presence of similar incomplete sequences in the corresponding regions of V. dahliae, V. longisporum, and V. albo-atrum isolates revealed a particular standard motif of insertions in the IGS region of the genus and is discussed. PMID- 11419484 TI - A simple in vitro model for growth control of bacterial biofilms. AB - The perfused biofilm fermenter was found to be unsuitable for the long-term culture and growth rate control of Staphylococcus aureus and Pseudomonas aeruginosa biofilms. In a simplified approach, biofilms of these organisms were grown within Sorbarod filter plugs which were perfused with culture medium. Pseudo-steady states were established which were stable over several days at which the growth rate of the biofilm was reproducible, measurable and significantly slower than in broth culture. Environmental scanning electron microscopy of dissected Sorbarods demonstrated an association of cells with the surfaces of individual cellulose fibres, and growth characteristic of biofilms. Relatively high cell numbers recovered from the Sorbarod model facilitated biochemical investigations of biofilm populations and cells released spontaneously from them. SDS-PAGE demonstrated significant differences between the protein profiles of biofilm and eluted populations, which include, in Staph. aureus, the repression of a 48 kDa protein and increased expression of a 21 kDa protein relative to planktonic controls cultured at equivalent growth rates. The paper demonstrates the suitability of the approach for the culture of biofilm samples which are suitable for biochemical analysis. PMID- 11419485 TI - Potentiation of a metabotropic glutamatergic response following NMDA receptor activation in rat hippocampus. AB - Interactions between metabotropic glutamate and N-methyl-D-aspartate (NMDA) receptor-mediated responses were investigated in hippocampal CA3 cells using the single electrode voltage-clamp method. Bath application (2.5-10 microM, 30 s) or iontophoresis of 1-amino-cyclopentyl-trans-1S,3R-dicarboxylate (ACPD), a selective agonist for metabotropic glutamate receptors, resulted in an inward current associated with a decrease in membrane conductance. Following transient bath application of NMDA (5-10 microM, 30-60 s), the ACPD-induced inward current was potentiated for a period of up to 25 min (by 61 +/- 8% with bath application, by 32 +/- 15% with iontophoresis). Transient application of NMDA did not result in a potentiation of ionotropic RS-alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) or metabotropic muscarinic responses. ACPD responses were not potentiated following transient AMPA application. Intracellular buffering of calcium with tetrapotassium bis(O-aminophenoxy)-ethane N,N,N',N'-tetraacetic acid (BAPTA) prevented potentiation by NMDA in all cells. Bath application of arachidonic acid did not mimic the NMDA-induced potentiation. These results demonstrate that activation of NMDA receptors can specifically induce a long-lasting potentiation of a metabotropic glutamatergic response in hippocampal CA3 pyramidal cells. The characterization of this interaction may contribute to the elucidation of the physiological significance of metabotropic glutamate receptors. PMID- 11419486 TI - Prescribing syringes to prevent HIV: a survey of infectious disease and addiction medicine physicians in Rhode Island. AB - This article describes the assessment of physicians' attitudes and practices regarding prescribing syringes to injection drug users (IDUs). A brief, anonymous, self-administered questionnaire was sent to all Infectious Disease and Addiction Medicine specialists in Rhode Island. Of 49 eligible physicians, 39 responded (response rate 80%). Most (95%) indicated that there is a legitimate medical reason for IDUs to obtain sterile syringes. Many (71%) agreed that they would prescribe syringes to prevent disease in IDUs if it were clearly legal to do so. We can conclude that physician syringe prescription to IDUs may be an acceptable supplement to existing HIV prevention strategies. PMID- 11419487 TI - Peer influences and susceptibility to smoking among California adolescents. AB - Using data from a diverse statewide sample of 10th-grade adolescents in California, in 1996-97, this study investigated the associations between peer influence variables and susceptibility to smoking. Peer influence variables included attitudes about the social consequences of smoking and subjective norms, as described by the Theory of Reasoned Action. Among never-smokers (N=2681) and ever-smokers (N=4248), attitudes about social consequences of smoking and subjective norms each were associated with an increased risk of susceptibility to smoking. The model explained a larger proportion of the variance in susceptibility among ever-smokers than among never-smokers. Results indicate that peer influences, including perceptions of the social consequences of smoking and perceived social norms, may make adolescents susceptible to smoking. PMID- 11419488 TI - Different forms of heroin and their relationship to cook-up techniques: data on, and explanation of, use of lemon juice and other acids. AB - Recent reports of the use of lemon juice in the preparation of heroin for injection have failed to recognize the importance of the different forms of heroin (in the form of the salt or the base) and the impact of this on the chemical manipulation required before injection. One hundred and four opiate addicts in London were interviewed about the forms of heroin (white, brown, etc) and their relationship to cook-up techniques (use of heat and acid). White heroin was typically prepared with water and heat; brown heroin was prepared with acid (citric acid or Vitamin C (ascorbic acid) or lemon juice) and heat; pharmaceutical heroin was prepared with water only (i.e. neither acid nor heat). On the last occasion of heroin use, brown heroin had been the form most commonly used, with over 90% of the sample using citric acid or vitamin C. Lemon juice was rarely used, and heat was almost universally applied in conjunction with lemon juice. PMID- 11419489 TI - Drug and alcohol offenders coerced into treatment: a review of modalities and suggestions for research on social model programs. AB - Outcome studies on drug and alcohol offenders coerced into treatment by the criminal justice system were reviewed. Positive outcomes were found for therapeutic community, methadone maintenance, and unspecified residential and outpatient programs. No outcome studies were found for court-mandated clients coerced into social model recovery programs. When developing studies, researchers should consider how clients perceive legal mandates and whether they are receiving pressures to enter treatment from other sources, such as family members, employers, friends or the welfare system. Use of the Social Model Philosophy Scale (SMPS) is suggested to correlate program characteristics and processes with outcome. PMID- 11419490 TI - Drinking and parental unpredictability among adult children of alcoholics: a pilot study. AB - Within-family differences that mediate the relationship between family history for alcoholism and alcohol consumption were investigated. Twenty adult children of alcoholics (ACOAs) and 20 children of social drinking parents (non-ACOAs) were interviewed in 1991 (mean age 24.2 years). Participants described family functioning while growing up and indices of their alcohol consumption. Family history for alcoholism had a direct association with two aspects of drinking. ACOAs reported more lifetime drinks and tended to report drinking more frequently during high school, as compared to non-ACOAs. ACOAs also reported more parental unpredictability, which in turn was associated with the above drinking indices. Finally, family history for alcoholism had an indirect association with drinking, as unpredictability mediated this relationship. Research and prevention implications are discussed. PMID- 11419491 TI - Comparison of the Rost and the CAGE alcohol screening instruments in young adults. AB - The Rost and CAGE alcohol screening instruments were compared in a random sample of 103 of 3609 young adults participating in a longitudinal panel. Rost (t = 6.01, df = 101, p<.001) and CAGE (t = 7.27, df = 97, p<.001) scores were significantly higher for those classified as having alcohol dependence than for those without dependence according to the short-form of the Composite International Diagnostic Inventory (SF-CIDI). The areas under the curve for the CAGE (0.840) and the Rost (0.783) were not statistically different (z = 1.30, p>.05). PMID- 11419492 TI - Sex-role strain in alcoholic women. AB - We investigated sources of sex-role strain between alcoholic and non-alcoholic women. We compared alcoholic (n = 55) and non-alcoholic (n = 51) women on the presence of strain in their sex-role identity and strain between sex-role identity, attitudes, and behaviors in 1996. One source of sex-role strain for alcoholic women was the discrepancy between their perceived femininity when intoxicated versus when sober. A second source of strain for alcoholic women was the discrepancy between their traditionally feminine sex-role identity as well as attitudes toward marital, parental, and educational roles and their participation in male-valued activities. We offer suggestions for future research. PMID- 11419493 TI - Substance use problems reported by college students: combined marijuana and alcohol use versus alcohol-only use. AB - This study examines the relationships among selected demographic characteristics, alcohol use only, alcohol plus marijuana use and substance-use related problems. Data are from 409 randomly selected undergraduate college students attending a large urban university. Poly-substance users were more likely than alcohol only users to be younger and experience all substance-use related problems studied. The relationship between poly-substance use and increased substance use associated problems was also identified when controlling for other common predictors of substance use problems. PMID- 11419494 TI - Late onset infection in very low birth weight infants in Malaysian Level 3 neonatal nurseries. Malaysian Very Low Birth Weight Study Group. AB - BACKGROUND: The purpose of this study was to examine the rate and mortality from late onset infection occurring in very low birth weight infants admitted to Malaysian nurseries. METHODS: Data on all infants 1500 g or below admitted to the 20 participating Level 3 nurseries were analyzed for late onset infection (clinical infection and positive blood or cerebrospinal fluid culture occurring after 48 h of life). RESULTS: The overall survival of the 962 study infants was 69%. The rate of late onset infection was 19.3%. The mortality in those with late onset infection was 30.8%. The most common infecting organism was Klebsiella pneumoniae, accounting for 38.3% of infections and 46.9% of deaths in infants with infection, followed by coagulase-negative staphylocci, 17.6 and 12.2%, respectively. On logistic regression analysis risk factors for late onset gram negative compared with gram-positive infection were endotracheal intubation at birth and blood transfusion. Hypoglycemia was associated with gram-positive infection. CONCLUSION: The late onset infection rate in Malaysian very low birth weight infants does not differ from that reported from developed countries, but the mortality is higher. This could be because of an excess of gram-negative infections. PMID- 11419495 TI - Strict interpretation of vaccination guidelines with computerized algorithms and improper timing of administered doses. AB - BACKGROUND: Frequently changing immunization recommendations may lead to incorrectly administered doses. OBJECTIVE: To determine the incidence and characteristics of inappropriately timed vaccinations. METHODS: Prospectively collected immunization histories of patients <5 years old from well-child care encounters with pediatric residents in a large urban clinic during a 3-month study period. New patients or those with no immunization history in the medical record were excluded. Paper records were verified before each visit and served as the immunization history. Immunization records were entered into and analyzed by the Massachusetts Immunization Information System with strict interpretation of minimum spacing and age guidelines to identify invalid vaccine doses. Reasons for invalidity were determined by manual review. Invalid doses were cross-referenced with clinic schedule to determine who delivered doses. RESULTS: Inclusion criteria were met by 690 encounters. Charts were available for review before the encounter for 580, containing 6983 total immunizations. Of these 289 (4.1%) administered doses were invalid; 206 of 580 (35.5%) patients had at least one invalid dose. Common invalid doses given were unnecessary poliovirus vaccine around 18 months (n = 66) and second hepatitis B vaccine given too soon after the first (n = 53). All types of providers gave invalid doses; pediatric residents and fellows delivered significantly more (P < 0.01). CONCLUSIONS: By strict interpretation of immunization guidelines, many patients were immunized incorrectly. Clinicians should be aware of common errors in vaccine dosing and national guidelines should be simplified. PMID- 11419496 TI - Observational study of the use of infection control interventions for Mycobacterium tuberculosis in pediatric facilities. AB - INTRODUCTION: Hospital transmission of Mycobacterium tuberculosis (TB) is a problem in US facilities where adults are treated. However, specific guidelines for facilities in which pediatric patients are cared for have never been defined, nor has any study attempted to assess pediatric health care worker (HCW) compliance with TB infection control (IC) guidelines. METHODS: An observational study was performed in two pediatric inpatient hospitals from May, 1996, to December, 1997. A trained observer tallied persons (i.e. professional HCWs, ancillary HCWs and non-HCWs) entering and leaving occupied TB isolation rooms and recorded adherence with IC practices (e.g. proper use of respirators, prompt door closures, door signage). RESULTS: Thirty children with confirmed or suspected TB were admitted during the study period and observed for a total of 242 h during which 656 visits by professional (n = 391) and ancillary (n = 131) HCWs and by family members (n = 134) were recorded. During 30% of visits doors remained open an average of 10 min, and during 20% of visits no respiratory protection was worn. In all, visitors wore the correct respiratory protection appropriately only 55% of the time. HCWs were more likely to wear respiratory protection when caring for children with a positive acid-fast bacillus smear than family members, but professional staff were no more likely than ancillary staff to do so. CONCLUSIONS: This is the first study to quantify compliance with IC practices for TB in pediatric hospitals. The majority of visitors entering TB isolation rooms occupied by children with confirmed or suspected TB complied with IC guidelines, but discrepancies were seen. Rather than relying on TB IC guidelines designed for adult facilities, guidelines specific for pediatric facilities that consider the local epidemiology of TB should be developed. PMID- 11419497 TI - Microbiology of otitis media in the Paris, France, area from 1987 to 1997. AB - PURPOSE OF THE STUDY: To investigate epidemiologic trends in the bacteriology of acute otitis media, data were recorded during a 10-year period by the same group of investigators during clinical studies in pediatric outpatients. METHODS: Bacterial samples were obtained before antibiotic treatment from 2149 children (age 3 to 36 months) with acute otitis media. All samples were transported, handled and cultured in the same way throughout the 10-year period. RESULTS: From the study patients 1862 samples of middle ear secretion were collected by tympanocentesis and 287 samples were collected from spontaneous otorrhea occurring within 24 h. Pathogens were isolated from 70% of patients. Pathogens included Haemophilus influenzae (40%), Streptococcus pneumoniae (31%) and Moraxella catarrhalis (8%). The incidence of beta-lactamase-producing H. influenzae was approximately 20% from 1987 to 1989, was approximately 35% from 1990 to 1995 and increased to 60 and 70% during the last 2 years. The rate of S. pneumoniae with decreased susceptibility to penicillin gradually increased from 7% in 1987 to 70% in 1996 and 1997. CONCLUSION: The very high incidence of antimicrobial resistant strains reached during a short period in pediatric acute otitis media in our area, emphasizes the need for frequent epidemiologic studies. PMID- 11419498 TI - Epidemiology of documented viral respiratory infections and acute otitis media in a cohort of children followed from two to twenty-four months of age. AB - BACKGROUND: Viral upper respiratory infections (URIs) are considered major risk factors for acute otitis media (AOM) in young children. We studied the epidemiology and relative roles of different viruses in respiratory infections in a cohort of 329 Finnish children followed from 2 months to 2 years of age. METHODS: A nasopharyngeal aspirate (NPA) was collected whenever the child had signs and/or symptoms of URI and tested for the presence of common respiratory virus antigens or infectivity/nucleic acid (only rhinoviruses). Possible repeated detections of a given virus during a 30-day period were considered to represent a single designated virus-specific episode. AOM and URI episodes were defined in a similar way. RESULTS: At least one virus was detected in 837 (41.7%) of the 2005 NPA specimens examined. Rates of URI and virus-specific episodes showed expected seasonal variation with major peak occurrences coinciding with or preceding those of AOM. The proportions of rhinoviruses, respiratory syncytial (RS) virus, parainfluenza virus (PIV) type 3, influenza virus A and adenoviruses were 63.1, 14.7, 6.7, 6.7 and 6.2% of the total of 761 virus-specific episodes. Influenza virus B, PIV1 and PIV2 were each responsible for approximately 1% of the episodes. AOM was diagnosed in 870 URI cases (43.4%) and in 43.3% of cases associated with a virus-positive NPA. The latter figure was clearly higher (57.7%) for RS virus-positive specimens. CONCLUSIONS: The seasonal coincidence of URI and AOM demonstrated the obvious role of URI in the pathogenesis of AOM. The occurrence of rhinoviruses and RS virus in URI was strikingly more common than that of any other virus tested. Although rhinoviruses were definitely the most frequently found viruses in NPA specimens, the association of RS virus with concurrent AOM was relatively higher than that of any other virus. PMID- 11419499 TI - Systemic nontyphoidal Salmonella infection in normal infants in Thailand. AB - BACKGROUND: The relative paucity of information about systemic nontyphoidal Salmonella (NTS) infection in infants without an underlying disease prompted this study. METHODS: Infants without an underlying disease, who had positive cultures for NTS from their normally sterile sites during 1978 through 1998, were included. Their medical records were reviewed. RESULTS: The study included 75 eligible infants; 68 (91%) had positive blood cultures. The spectrum of disease included transient bacteremia (5), bacteremia without localized infection (37), bone and joint infection (5) and meningitis (28); 53 and 88% of infants were < or =3 and < or =6 months old, respectively. All infants with localized infection were < or =7 months old, and infants with meningitis were 3.35+/-1.87 (mean +/- SD) months old. In bacteremic infants risks for localized infection and meningitis were 30 and 24%, respectively. Abnormal neurologic findings were the only predictor for meningitis. Relapse of meningitis occurred in two infants despite treatment with cefotaxime for 4 and 6 weeks. Severe neurologic deficit occurred in 21% of infants with meningitis. Of 11 infants with meningitis who received early treatment at this tertiary care center, 1 died but none had severe neurologic deficits; whereas of 17 referred cases, 2 died and 6 had severe neurologic abnormalities. No death occurred in infants without meningitis. CONCLUSION: Systemic NTS infection in normal infants has a variable disease spectrum. Infants 0 to 6 months of age are at high risk for localized infection, especially meningitis. PMID- 11419500 TI - Early responses to nonconjugated polyribosylribitol phosphate challenge as evidence of immune memory after combined diphtheria-tetanus-pertussis-polio Haemophilus influenzae type b primary vaccination. AB - OBJECTIVES: A high risk of invasive Haemophilus influenzae type b (Hib) disease exists in the first few years of life. A reduction in anti-polyribosylribitol phosphate (PRP) antibody concentrations follows the administration of certain diphtheria-tetanus-acellular pertussis (DTPa)-based Hib conjugate combined vaccines. However, these combined vaccines prime the immune memory, which is an important factor in protection. As yet there is no direct evidence of the time scale involved in the development of the immune memory post-primary vaccination. In this report we investigated the presence of immune memory at 10 and 12 months of age, 4 and 6 months after primary vaccination of young infants with a pentavalent combination of DTPa, inactivated poliovirus vaccine (IPV) and Hib (DTPa-IPV/Hib) vaccine. METHODS: In two trials (A and B) infants received DTPa IPV combined with Hib-tetanus conjugate (PRP-T) vaccine at 2, 4 and 6 months of age. The presence of immune memory was assessed by measuring anti-PRP concentrations 7 to 10 days after a nonconjugated PRP challenge given at 10 months in Trial A and at 12 months in Trial B. RESULTS: Administration of a nonconjugated PRP challenge 4 and 6 months after primary vaccination in Trials A and B, respectively, elicited an increase in anti-PRP geometric mean concentrations (4.5 and 5.8 microg/ml, respectively) within 7 to 10 days. These concentrations exceed those reported in the literature involving unprimed children who had received a single dose of nonconjugated PRP at the same age. CONCLUSION: The results demonstrate the development of anti-PRP immune memory at an early age, 4 and 6 months after completion of a three dose primary vaccination course of combined DTPa-IPV/Hib vaccine. The ability of primed infants to mount a rapid response is an important observation given the high risk of Hib infection at this critical age. PMID- 11419501 TI - Predictors of ventriculoperitoneal shunt pathology. AB - BACKGROUND: Diagnosis of ventriculoperitoneal (VP) shunt pathology remains a dilemma in patients with nonspecific constitutional signs and symptoms. Eosinophilia has been described in association with shunt infection and malfunction. Our purpose was to further define the relationship of eosinophilia and shunt pathology and to determine other predictors of VP shunt infection and malfunction. METHODS: Records of all patients admitted with a suspected VP shunt infection or malfunction were reviewed. The following data were abstracted: age; reason for and age at initial shunt placement; number of revisions; date of last revision; history of fever or vomiting; ventricular fluid cell count; differential and culture; complete blood count and differential; need for shunt revision or replacement; and use of antibiotics. After exclusion of patients admitted for initial shunt placement, the remainder were divided into three groups: those with shunt infection; those with shunt malfunction; and those without documented infection or malfunction. RESULTS: Of 12 patients with shunt infection and 69 with shunt malfunction, 2 and 11, respectively, had eosinophilia defined as > or =5%. The presence of eosinophilia had a 96% positive predictive value for shunt pathology and raised the pretest probability of pathology from 84% to a post test probability of 96%. The combination of fever history and ventricular fluid neutrophils >10% had a 99% specificity for shunt infection, had a 93 and 95% positive and negative predictive value, respectively, and raised the pretest probability of infection from 12% to a posttest probability of 92%. CONCLUSIONS: In patients suspected of having a VP shunt malfunction, the presence of > or =5% eosinophils in the ventricular fluid indicates shunt pathology. The combination of fever and ventricular fluid neutrophils > 10% is predictive of shunt infection. PMID- 11419502 TI - Predictive factors associated with significant urinary tract abnormalities in infants with pyelonephritis. AB - BACKGROUND: Major urinary tract abnormalities are detected in 20 to 40% of infants with acute pyelonephritis (APN). Early detection of structural defects is essential for protecting the kidneys from reinfection and subsequent scarring. The purpose of this study was to investigate whether any factors present during the acute phase of infection could predict the presence of existing significant urinary tract abnormalities in infants. METHODS: A prospective study of 180 infants, aged 1 to 24 months, with APN was conducted. Blood and urine samples were collected. Renal ultrasound (US) was performed within 0 to 6 days from admission. Final diagnosis of the urinary tract anatomy was elucidated using the results of two or more radiologic imaging studies. RESULTS: Risk factors for the presence of significant urinary tract abnormalities in infants were pathogens other than Escherichia coli in urine [relative risk (RR) 3.4, 95% confidence interval (CI) 2.2 to 5.3; P = 0.001], positive blood culture (RR 2.3, 95% CI 1.3 to 4.0; P = 0.039), young age (1 to 6 months) (RR 2.2, 95% CI 1.3 to 3.9; P = 0.004), lack of papG adhesin genes of E. coli in urine (RR 2.1, 95% CI 1.2 to 3.9; P = 0.016) and abnormal renal US (RR 2.0, 95% CI 1.2 to 3.4; P = 0.008). CONCLUSIONS: Infants 1 to 6 months of age with APN caused by bacteria other than E. coli or by papG-negative E. coli strain, positive blood culture and abnormal renal US carry an increased risk for significant urinary tract abnormalities and need enforced follow-up. PMID- 11419503 TI - Mother-to-child transmission of the human immunodeficiency virus in Texas. AB - BACKGROUND: The Pediatric Spectrum of HIV Diseases (PSD) project has been collecting data on HIV-exposed children in Texas since 1989. These data have now been analyzed to describe mother-to-child transmission in Texas and to provide much needed information on the magnitude of the pediatric HIV epidemic in the state. METHODS: We examined trends in the numbers of perinatally exposed children and perinatally acquired cases of HIV in the Texas PSD cohort. We calculated transmission rates and relative risks for 656 children born from January, 1995, to July, 1998, that received all or part of the ACTG 076 regimen. RESULTS: Only a small proportion (38%) of pairs of an HIV-infected mother and her HIV-exposed child received the full AIDS Clinical Trial Group 076 (ACTG 076) regimen; only 73% of the mothers received at least some prenatal care. In recent years, however, the numbers of perinatally exposed children and perinatally acquired cases of HIV have decreased in Texas. Univariate analyses showed that a reduction in the vertical transmission of HIV was associated with receipt of a full ACTG 076 regimen, receipt of a partial ACTG 076 regimen and residence in Dallas County. CONCLUSIONS: Findings identify a gap in meeting the health care needs of pregnant HIV-infected women and suggest missed opportunities to prevent mother-to child transmission of HIV. At the same time this study confirms progress in prevention efforts to reduce mother-to-child transmission of HIV in Texas. PMID- 11419504 TI - Bacterial etiology of acute myringitis in children less than two years of age. AB - BACKGROUND: Acute myringitis is an inflammation of the tympanic membrane that occurs alone or in association with external otitis or otitis media. The two clinical entities, based on physical examination, are bullous myringitis and hemorrhagic myringitis. OBJECTIVES: To investigate the association of concomitant middle ear disease with acute myringitis and to analyze the bacteriologic findings of the middle ear fluid in children with acute myringitis. METHODS: A prospective longitudinal cohort study of 2028 children age 7 to 24 months at primary care level in the Finnish Otitis Media Vaccine Trial. Matched case control design for analysis of bacterial pathogen distribution. RESULTS: There were 82 children in whom 92 ears were diagnosed with acute bullous myringitis and 37 children in whom 40 ears were diagnosed with hemorrhagic myringitis during the follow-up. Middle ear disease was associated with bullous myringitis in 97% of ears and with hemorrhagic myringitis in 82% of ears. Bacterial pathogen distribution was similar to that of acute otitis media, although a higher proportion of Streptococcus pneumoniae was detected in both bullous and hemorrhagic acute myringitis. CONCLUSIONS: Middle ear fluid was present in vast majority of ears with acute myringitis in young children. The same etiologic bacteria were found in acute myringitis as in acute otitis media, but S. pneumoniae was the major pathogen. Acute bullous myringitis should be treated as acute otitis media in children <2 years of age. PMID- 11419505 TI - Nosocomial acquisition and transmission of antibiotic-resistant gram-negative organisms in the pediatric intensive care unit. PMID- 11419506 TI - Update on varicella. PMID- 11419507 TI - Vancomycin-resistant enterococci in the pediatric patient. PMID- 11419508 TI - Successful treatment of Acanthamoeba meningitis with combination oral antimicrobials. AB - Acanthamoeba was implicated as the causative agent of chronic meningitis in three apparently immunocompetent children. Diagnosis was established by cerebrospinal fluid wet mount examination and culture. Two children improved rapidly with combination oral therapy composed of trimethoprim-sulfamethoxazole, rifampin and ketoconazole. PMID- 11419509 TI - Safety and tolerance of palivizumab administration in a large Northern Hemisphere trial. Northern Hemisphere Expanded Access Study Group. AB - An Expanded Access Study was conducted to collect additional safety data on palivizumab. Preterm infants with or without bronchopulmonary dysplasia received palivizumab every 30 days during the respiratory syncytial virus season. Adverse events were low (6.9%) in the 565 subjects. Serious adverse events included hospitalization and 1 case of respiratory syncytial virus bronchiolitis not requiring hospitalization. This study reaffirms the safety and tolerability of palivizumab. PMID- 11419510 TI - Concomitant aseptic meningitis and bacterial urinary tract infection in young febrile infants. AB - We evaluated the incidence and implications of coexistent bacterial urinary tract infection and aseptic meningitis in 1629 young febrile infants (age 1 to 60 days) who underwent sepsis work-up. Urinary tract infection was diagnosed in 13.2% and aseptic meningitis in 8.8%. Eleven patients (0.7%) had both infections. In view of possible coinfection initial laboratory results may be insufficient for decision-making regarding treatment in young febrile infants. PMID- 11419511 TI - Treatment of Calmette-Guerin bacillus adenitis: a metaanalysis. AB - Various treatments have been used to decrease the risk of suppuration; the most troublesome complication of Calmette-Guerin bacillus adenitis, but results are controversial. Metaanalysis of four randomized controlled trials revealed no significant difference in the frequency of suppuration between the treatment and control groups for all treatments [relative risk (RR), 1.10; 95% confidence interval (CI), 0.88 to 1.38], erythromycin (RR 1.04; 95% CI 0.79 to 1.37) and isoniazid (RR 1.35; 95% CI 0.84 to 2.18). Therefore medical treatment does not reduce the frequency of suppuration in Calmette-Guerin bacillus adenitis. PMID- 11419512 TI - Fatal disseminated intravascular coagulation caused by Mycoplasma pneumoniae. AB - We report fatal disseminated intravascular coagulation caused by Mycoplasma pneumoniae infection in a 7-year-old white boy. The diagnosis was based on relevant clinical material, absence of any other infections, a 4-fold rise in titer of complement fixation for M. pneumoniae and open lung biopsy findings. We discuss the association between M. pneumoniae and autoimmune mechanism. PMID- 11419513 TI - Response of refractory Kawasaki disease to pulse steroid and cyclosporin A therapy. AB - We describe a child with aggressive and protracted Kawasaki disease with coronary aneurysms, myocarditis, pericarditis and valvular insufficiency, despite repeated administration of intravenous immunoglobulin. After a transient response to pulse corticosteroids, his disease ultimately subsided with combination therapy with pulse and high dosage corticosteroids and cyclosporin A. Aggressive immunosuppressive therapy with high dosage corticosteroids and cyclosporin A may be beneficial in patients with refractory Kawasaki disease. PMID- 11419514 TI - Thyroiditis as a presenting feature of mumps. AB - Thyroiditis complicating mumps is rare and occurs 1 week after the parotitis. A 9 year-old boy with a history of contact with a case of mumps presented with thyroid swelling. Thyroid scan showed a diffusely reduced uptake. The aspiration cytology showed lymphocytic thyroiditis. Thyroid function tests were normal and antithyroid antibodies were absent. Parotitis occurred 12 days after the onset of thyroiditis. PMID- 11419515 TI - Urinary findings in asymptomatic subjects with spina bifida treated with intermittent catheterization. AB - The urines from 43 asymptomatic children with spina bifida were examined. Eighty one percent were abnormal because of bacteriuria and pyuria (51%), bacteriuria alone (26%) or pyuria alone (5%). Interleukin-8 was elevated in 54% of the abnormal urines. The presence of pyuria and interleukin 8 suggests that the asymptomatic bacteriuria reflects low grade infection rather than colonization. PMID- 11419516 TI - Follow-up to the statewide model detection and prevention program for geriatric alcoholism and alcohol abuse. AB - A follow-up study was conducted to investigate the long term consequences of a statewide geriatric alcoholism detection and prevention effort in Virginia. High levels of knowledge had been retained seven years after the initial training. Quantitative and qualitative data provided by 64 of the volunteers, who were trained to conduct workshops in their communities, verified the program's ability to: 1) encourage further training or study in the area, 2) enhance the detection of alcohol problems, and 3) increase the incidence of assistance to individuals with alcohol problems. Ancillary data revealed the program's effects on personal drinking habits. PMID- 11419517 TI - An exploratory analysis of racial factors in staff burnout among assertive community treatment workers. AB - We examined racial differences in burnout among case managers working with people with severe mental illness, using the Maslach Burnout Inventory. Compared to Caucasians, African Americans reported significantly less Emotional Exhaustion and Depersonalization, but did not differ on levels of Personal Accomplishment. These differences could not be explained by geographic location or perceptions of the work environment; however, age accounted for group differences in Depersonalization. Racial incongruence with caseload appeared to be one factor in burnout, particularly for Emotional Exhaustion. Race of both staff and clients may be important to consider in understanding staff burnout. PMID- 11419518 TI - Negative and supportive social interactions and quality of life among persons diagnosed with severe mental illness. AB - This study examined the relative importance of negative and supportive social interactions in predicting different aspects of quality of life (QOL) in a sample of persons diagnosed with severe mental illness (n = 104). Controlling for other variables that might explain such a relationship, we found that negative social interactions were significantly related to lower QOL in three subjective domains, while supportive social interactions were related to higher QOL in four objective domains and one subjective domain. We found negative social interactions that are stigmatizing to be particularly important in predicting QOL; additional analyses suggested that perceived stigma partially mediated the relationship between negative social interactions and QOL. We discuss the implications of the present findings for the study of the link between social relationships and QOL among persons with mental illness. PMID- 11419519 TI - Barriers to help seeking for mental disorders in a rural impoverished population. AB - This study examined barriers to seeking mental health care reported by individuals in a rural impoverished population, by screening 646 randomly selected adults for depression, anxiety, and alcohol abuse. Respondents who screened positive were randomly assigned to one of three groups: (1) no intervention, (2) an educational intervention alone, or (3) the educational intervention in the presence of a significant other. Those who screened positive for disorders cited barriers to care at significantly higher rates than respondents who screened negative. Respondents who received the educational intervention endorsed several barriers at significantly lower rates in the follow up telephone call (subsequent to the intervention) than in the original interview (prior to the intervention). Virtually all respondents in a subsample of 142 subjects (99.3%) said they would seek mental health care if they thought mental health services would help them. PMID- 11419520 TI - System effects of the Kansas Mental Health Reform Act of 1991. AB - This paper analyzes the impact of the Kansas Mental Health Reform Act on client outcomes. The Act is of general interest because it reflects a trial of greater accountability without major changes in financial incentives. It made Community Mental Health Centers [CMHCs] gatekeepers that were accountable for services for adults with severe and persistent mental illnesses. The Act sought reductions in hospitalizations rates, expanded use of community support services, and increased independent living. The structure of the Act and Client Status Reports allow rigorous examination of these outcomes. The number of clients served increased significantly. Even though hospital days fell by 23%, there is no clear evidence that the Act itself reduced hospital days per client. The proportion of community support program clients residing independently rose significantly; the proportion participating meaningfully in the labor market fell. The goals of the Act were realized overall, but the performance of CMHCs varied considerably. PMID- 11419521 TI - Telepsychiatry in the heartland: if we build it, will they come? AB - The quality of care provided by telemedicine and its acceptability to persons who live in rural areas is largely undetermined. In this study, service satisfaction and functional status in persons using telemedicine was compared to those receiving face-to-face services at two rural sites over a 2-year study period. Similar ratings of satisfaction and clinical status were observed in twelve patients who received services under both modalities. Although there are many obstacles to successful program implementation, telepsychiatry appears to offer an acceptable and adequate alternative mode of service delivery to persons who live in rural areas. PMID- 11419522 TI - Beyond the crisis. PMID- 11419523 TI - Letter across the Pacific. PMID- 11419524 TI - Intervention without interference: a column of Befrienders International. PMID- 11419525 TI - The end of suicidology. PMID- 11419526 TI - Serotonin disturbances and suicide risk: is aggression or anxiety the interjacent link? PMID- 11419527 TI - Sleep disorder, depression, and suicidality in female sexual assault survivors. AB - The role of sleep in psychiatric illness in general, and depression and suicidality in particular, is poorly understood and has not been well researched despite the pervasiveness of sleep complaints in these conditions. As an exploratory, hypothesis-generating study, female sexual assault survivors with posttraumatic stress disorder (n = 153) who had enrolled in a nightmare-treatment program were assessed for subjectively determined sleep breathing and sleep movement disorders. Diagnoses of potential disorders were based on clinical practice parameters and research algorithms from thefield of sleep disorders medicine. Potential sleep breathing and sleep movement disorders were present in 80% of the participants (n = 123) and included three subgroups: sleep-disordered breathing only (n = 23); sleep movement disorder only (n = 45); and both sleep disorders (n = 55). Based on the Hamilton Depression Rating Scale and Suicide subscale, participants with potential sleep disorders suffered greater depression (Cohen's d = .73-.96; p < .01) and greater suicidality (Cohen's d = .57-.78; p < .05) in comparison to participants without potential sleep disorders. The group with both sleep disorders suffered from the most severe depression and suicidality. A provisional hypothesis is formulated that describes how sleep disorders may exacerbate depression and suicidality through the effects of chronic sleep fragmentation. PMID- 11419528 TI - Epidemiology and psychiatric morbidity of suicidal ideation among the elderly. AB - This study investigates the epidemiology and psychiatric morbidity of the wish to be dead, suicidal ideation, and suicidal intent in a group of elderly persons (> 70 years). A representative community sample of 516 persons aged 70 to 105 was extensively investigated by psychiatrists using the structured interview GMS-A and various other self-rating and observer-rating scales. Diagnoses were made according to DSM-III-R and clinical judgment. In a cross-section of this population, we found the following prevalence rates: At the time of the study, 14.7% of the elderly community had symptoms of tiredness of life, 5.4% wished to die, and 1% showed suicidal ideation or gestures. Depending on the intensity of suicidality, 80% to 100% were clinically diagnosed as suffering from psychiatric disorders and 50-75% showed symptoms fulfilling the criteria of at least one specific psychiatric diagnosis. Further, logistic regression analysis showed a significant influence of major depression and specific DSM-III-R diagnosis on suicidality in old age. Our conclusion is that suicidal ideation in the elderly is usually a sign of a mental illness warranting diagnosis and treatment rather than assisted suicide. PMID- 11419529 TI - Pre-Durkheim suicidology. AB - Durkheim is generally regarded as the founder of the scientific study of suicide. However, even a cursory review of 18th- and 19th-century literature reveals an increasingly sophisticated scientific approach to suicide, culminating in the encyclopedic research of Morselli in 1879 and the critical review of Tuke in 1892, works that lose nothing in comparison with Durkheim's Le Suicide of 1897. This review, while in no way drawing Durkheim's role as a founder of scientific sociology into question, indicates that his position in regard to the study of suicide does warrant reconsideration. PMID- 11419530 TI - The internet, suicide, and suicide prevention. PMID- 11419531 TI - Proceedings of the international meeting "Suicide: interplay of genes and environment". PMID- 11419532 TI - A potentially fatal effect of topical anesthesia. PMID- 11419533 TI - Bradycardia with a changing P-R interval. PMID- 11419534 TI - Advances in managing Parkinson's disease. AB - The medical choices are now more numerous: Dopamine agonists may have value not only for addressing the motor fluctuations of levodopa therapy but also for deferring the use of levodopa. Surgical choices are bolstered by the addition of investigational options such as neuronal transplantation, in addition to options currently approved, such as pallidotomy. PMID- 11419535 TI - Antiaging therapy: an overview. AB - Today's researchers are exploring caloric restriction, cell-based therapies, hormonal therapies, and genetic manipulations. So far, caloric restriction has the soundest basis, and estrogen replacement is among the interventions most widely used. As the human genome is studied, treatments with genetic mechanisms move all the closer to becoming reality. PMID- 11419536 TI - Medical issues in treating renal transplant patients. AB - As the survival of renal transplant patients has increased, so has their risk of significant morbidity. Immunosuppressive drugs can exacerbate preexisting conditions and promote development of new disease. Effective long-term care of transplant recipients requires an understanding of how these drugs affect clinical evaluations and treatments. PMID- 11419537 TI - Finding and treating subclinical hypothyroidism. PMID- 11419538 TI - Effect of UV irradiation on type I collagen fibril formation in neutral collagen solutions. AB - BACKGROUND: Collagens have the well-known ability to spontaneously self-associate to form fibrils at physiological temperature and neutral pH in vitro and in vivo. Because solar UV may photochemically alter collagen, the kinetics of fibril formation may be modified. Thus, we have begun a systematic study of the effect of various UV wavebands on fibril formation. METHODS: Citrate-soluble calf skin collagen (Elastin Products) was dissolved at 0.05% in 0.5 M HOAc, dialyzed over 2 days into two changes of 0.0327 M phosphate buffer, pH 7.0 at 4 degrees C, and centrifuged at 48,000 x g. Photolysis was carried out at 4 degrees C with either (a) UVC (UVG-11 lamp), (b) filtered solar-simulating radiation (SSR) or UVA (SSR or UVL-21 lamp filtered with a 2.0 mm Schott WG 345 filter). Gelation was commenced by rapidly raising the temperature from 8 degrees C to 33 degrees C. Nucleation and growth were followed by turbidimetric measurements at 400 nm. RESULTS: UVC radiation (0-17.3 J/cm2) resulted in a dose-dependent decrease in the rate of fibril growth. Under these conditions, concomitant collagen crosslinking and degradation occurred. Fibril nucleation, a prerequisite for growth, was rapid (threshold approximately 2 min) and was not affected by UVC, UVA or SSR. SSR (0-1,320 J/cm2) caused a small decrease in growth rate and in the degree of fibril formation. UVA radiation (0-1,080 J/cm2) had a similar effect. "Direct" photochemical damage thus paralleled absorption via various collagen chromophores, with UVC>SSR approximately UVA. The presence of riboflavin (RF) resulted in groundstate interactions that markedly altered both nucleation and growth kinetics. Irradiation with 29.6 J/ cm2 UVA in the presence of RF photosensitizer caused relatively minor additional changes in fibrillation kinetics. CONCLUSIONS: These results collectively indicate that fibril formation is markedly dependent on specific ground state interactions and relatively insensitive to nonspecific UV damage. On the other hand, fibrils thus formed from photochemically altered collagen may have altered structural properties that could have subtle but unfavorable effects on the local dermal milieu in vivo. Notwithstanding, the relative insensitivity of fibrillogenesis to non-specific photochemical damage probably represents a favorable adaptation, overall, which tends to conserve the mechanical integrity of the skin. PMID- 11419539 TI - UVB-induced GM-CSF production is suppressed by dexamethasone in HaCaT cells. AB - BACKGROUND: Epidermal keratinocytes are important sources of a wide variety of cytokines that include the Granulocyte-macrophage Colony Stimulating Factor (GM CSF). Glucocorticoids have been shown to inhibit the production of several cytokines. However, their effect on GM-CSF synthesis by keratinocytes is still unknown. METHODS: The effects of glucocorticoid on GM-CSF production by keratinocytes were evaluated using ELISA and RT-PCR analysis. RESULTS: GM-CSF secretion by HaCaT cells increased with increasing UVB exposure. Dexamethasone suppressed basal release of GM-CSE In addition, it strongly inhibited both the UVB-mediated augmentation of GM-CSF protein production and mRNA expression. Lincomycin enhanced slightly the inhibitory effect of dexamethasone on GM-CSF synthesis, while lincomycin itself had no effect on GM-CSF secretion. CONCLUSION: Results showed that dexamethasone suppressed basal release and UVB-induced production of GM-CSF by keratinocytes. PMID- 11419540 TI - Ultraviolet-filtering properties of commonly used tissue cell culture plasticware. AB - BACKGROUND: Fluorescent sunlamps are a common source of ultraviolet radiation (UVR) for photobiology research. However, these lamps emit a significant amount of biologically "irrelevant" wavelengths that, if not removed, can drastically skew results and perhaps lead to mistaken conclusions regarding human photobiology. The use of a cellulose triacetate sheet (Kodacel) to filter the shorter ultraviolet wavelengths has become the accepted standard in photobiology. Over time, the transmission characteristics of this sheet may be altered due to photochemical changes. In addition, in vitro experiments utilizing filtered fluorescent sunlamps require the removal of plastic tissue cell culture lids, increasing the possibility of contamination. METHODS: We evaluated the transmission characteristics of various commercially available plastic lids used in tissue cell cultures. In addition, we used a biological system containing the human elastin promoter/chloramphenicol acetyltransferase reporter gene construct to compare the effects of filtering from these plastic lids. RESULTS: Here, we demonstrate that the transmission of UVR and the biological response through plastic culture dish lids is similar to that of Kodacel. CONCLUSION: Although this is an improvement for in vitro experiments, further improvements can be made using more realistic UVR sources, e.g. UVA-340 lamps, which mimic the short wavelengths of sunlight. PMID- 11419541 TI - Usage of real-time ultraviolet radiation data to modify the daily erythemal exposure of primary schoolchildren. AB - BACKGROUND/AIMS: Primary schoolchildren in their everyday school life are exposed to solar ultraviolet radiation. This may be through time spent outdoors whilst having meal breaks, physical education classes and other class orientated outdoor activities. METHODS: This research investigates the UV exposure of primary schoolchildren and the effect real-time UV irradiances data and an associated software package, UVGUIDE, have on UV exposure. This software utilises scientifically collected data, such as facial distribution of UV, as well as accessing real-time on-line UV irradiances data to estimate the UV distribution to the head region. The students can also enter other parameters such as hat usage and hat type to show the effect of using such a UV protective device on their facial UV distribution. CONCLUSIONS: The results from this study found that the average 3-day erythemal UV exposure in late summer and early autumn to the left shoulder was 33% higher for the students not having access to the on-line UV data and software package. PMID- 11419542 TI - What's new in photoimmunology. PMID- 11419543 TI - The pathophysiology of photosensitivity in lupus erythematosus. AB - The strong association between photosensitivity and lupus erythematosus has led to the suggestion that abnormal photoreactivity participates in the pathogenesis of cutaneous lesions. In this review we discuss the evidence for abnormal cutaneous reactivity to sunlight in lupus and speculate on the cellular, molecular and genetic factors that may underlie this abnormality. PMID- 11419544 TI - Treatment of surgical patients is evidence-based. AB - OBJECTIVES: To assess the type of evidence that supported our management of surgical patients. DESIGN: Retrospective audit. SETTING: Teaching hospital, Republic of Ireland. PATIENTS: All 222 patients admitted by two surgical teams during one month. Patients admitted for diagnostic procedures were included if they were treated as a result. MAIN OUTCOME MEASURES: Diagnosis and treatment were established from the notes. Evidence for each intervention was then sought in MEDLINE 1986-1999, Best Evidence 1991-1999, and the Cochrane Database of Systematic Reviews Issue 3, 1999, and graded into one of five categories: systematic review, meta-analysis, randomised controlled trial, prospective study, or retrospective study. RESULTS: Treatment was supported by systematic review in 14, by meta-analysis in 12, by randomised controlled trial in 75, by prospective studies in 55, by retrospective studies in 58, and by no evidence in 8. CONCLUSIONS: Everyday surgical practice in our unit is supported by good quality evidence from recent publications. PMID- 11419545 TI - Abdominal injuries caused by bicycle handlebars. AB - OBJECTIVE: To find out whether the increase in the number of children admitted with injuries from mountain bicycle handlebars is attributed to recent changes in the design of children's bicycles. DESIGN: Retrospective study. SETTING: Teaching general hospital, Israel. SUBJECTS: 76 children who presented with abdominal injuries caused by bicycle handlebars. RESULTS: In 12 of the 76 children, there was an imprint of the handlebar edge on the hypochondrium. The most common injuries were isolated ruptures of spleen or liver, (14 and 11 patients, respectively). Five of the 25 patients were operated on and the rest treated conservatively. CONCLUSIONS: Children with an imprint or bruise made by the handlebar edge on the abdominal wall, or who give a clear history of injuries by a bicycle handlebar should be treated with great care. BMX handlebars are relatively high (for young riders) and wide; they also turn freely and are therefore in direct line with the upper abdomen. Prohibiting the use of bicycles with unpadded handlebars may prevent some of these intra-abdominal injuries. PMID- 11419546 TI - Diagnosis and treatment of isolated tuberculous mediastinal lymphadenopathy in adults. AB - OBJECTIVE: To evaluate the diagnostic yield of bronchoscopy and mediastinoscopy in adults with isolated mediastinal tuberculous lymphadenitis and to assess the effect of antituberculous treatment. DESIGN: Prospective longitudinal cohort study of 34 patients with mediastinal tuberculous lymphadenitis followed for 6 to 19 months after completion of treatment. SETTING: Tertiary care hospital, Kuwait. PATIENTS: 34 consecutive patients who presented with isolated mediastinal lymphadenopathy from 1996 to 1998. INTERVENTIONS: Bronchoscopy and cervical mediastinoscopy for all patients. MAIN OUTCOME MEASURES: Diagnostic yield of bronchoscopy and mediastinoscopy, and the outcome of treatment in patients with tuberculous lymphadenopathy. RESULTS: The mean age was 35 years (range 15-58). The most common symptoms were cough, fever, and weight loss. The chest radiographs and computed tomograms showed abnormal mediastinal shadows with no evidence of parenchymal disease. All patients had right sided paratracheal lymphadenopathy. Tuberculin skin test gave a weal of >15 mm in 17 patients (50%). Sputum smears and cultures failed to grow acid-fast bacilli in any patient. Seven patients had an endobronchial abnormality and samples taken at bronchoscopy gave a definite diagnosis in 3 (9%). Paratracheal lymph node biopsy and culture by mediastinoscopy diagnosed tuberculosis in all cases. All patients were treated by a six month course of rifampicin and isoniazid supplemented initially by pyrazinamide for two months. Twenty-eight patients had a good response and the remaining patients were treated for a further 3 months. CONCLUSIONS: Bronchoscopy has a low diagnostic yield in mediastinal tuberculous lymphadenopathy in the absence of a parenchymal lesion. Mediastinoscopy is a safe but invasive procedure and provides a tissue diagnosis in most cases. Six months treatment with rifampicin and isoniazid supplemented initially by pyrazinamide is adequate treatment for most adults with tuberculous mediastinal lymphadenopathy. PMID- 11419547 TI - Familial papillary carcinoma of the thyroid: report of three families. AB - OBJECTIVE: To report our experience of the clinical characteristics, treatment and outcome of familial papillary carcinoma. DESIGN: Descriptive study. SETTING: Tertiary referral centre, Spain. PATIENTS: Eight patients from three families. INTERVENTIONS: Treatment for familial papillary carcinoma of thyroid. MAIN OUTCOME MEASURES: Age, sex, reason for consultation, preoperative investigation, indication for operation and type, variety of papillary carcinoma and characteristics (size, adenopathies, multicentricity and bilaterality), treatment with 131I, and outcome. RESULTS: The mean (SD) age was 46 (14) years, and seven were women. In all but one patient, who was admitted for hypercalcaemia and operated on for primary hyperparathyroidism, cervical palpation and ultrasound showed a thyroid nodule. Needle aspiration suggested papillary carcinoma in 6/7, who were all treated by total thyroidectomy. Four patients with enlarged nodes also had bilateral neck disection. All the papillary carcinomas were well differentiated, mean (SD) diameter was 1.5 (1) cm and in four were microcarcinomas. Three patients presented with invaded nodes. They were all treated postoperatively with 131I, except the patient with hyperparathyroidism who had a microcarcinoma of 0.5 cm. After a median (range) follow-up of 1 (0, 6 7) years the patients are well, except one had a high thyroglobulin concentration seven years later; however clinical exploration and morphological screening were within normal limits and he was treated with 131I 5.6 x 10(9) Bq. CONCLUSIONS: About a third of familial papillary carcinomas of the thyroid have lymph node involvement at the time of diagnosis, and have a relatively good prognosis if treated in accordance with the stage of the disease. PMID- 11419548 TI - Value of 99mTc-MIBI scintimammography in women with impalpable breast lesions seen on mammography. AB - OBJECTIVE: To find out if 99mTc-methoxyisobutylisonitrile scintimammography (MIBI) is useful in the evaluation of impalpable breast lesions in women referred for biopsy after mammography screening. DESIGN: Observational and prospective study. SETTING: Radiological screening campaign in Spain. PATIENTS: 36 patients referred between September 1997 and June 1998 (37 biopsies) with impalpable breast lesions referred consecutively from a screening campaign. Ages ranged from 50 to 64 years. INTERVENTIONS: Mammography, ultrasound, MIBI, and wire-guided biopsy. MAIN OUTCOME MEASURES AND RESULTS: Mean values for sensitivity, specificity, positive predictive value, and negative predictive value for scintimammography were 78.9, 72.2, 75, and 76.5 respectively (p = 0.002). The logistic regression analysis showed that 99mTc-MIBI scintimammography result predicted (p = 0.003) the risk of breast cancer in this group of patients (odds ratio: 9.75), particularly infiltrating ductal carcinoma. CONCLUSIONS: MIBI scintimammography in patients referred for biopsy after mammography screening does not so much rule out cancer as confirm the diagnosis. Its practical benefit would be not so much avoiding biopsy for these patients as aiding in planning the radicality of their excision. PMID- 11419549 TI - Relationship between circulating high density lipoprotein concentrations and interleukin-6 release during abdominal operations. AB - OBJECTIVE: To investigate whether circulating high density lipoprotein (HDL) concentrations influence inflammatory cytokine release during abdominal operations. SETTING: University Hospital, Japan. PATIENTS: 30 patients who underwent curative operations for gastric cancer. MAIN OUTCOME MEASURES: Serum HDL and cytokine concentrations were measured preoperatively, and portal and systemic blood samples were taken for the analysis of cytokine and endotoxin concentrations during operations. RESULTS: Portal blood interleukin-6 (IL-6) values of patients with low serum HDL concentrations (n = 6) (< 0.9 mmol/L) were significantly (p < 0.01) higher than those in patients with normal or high HDL concentrations (n = 24) (> or = 0.9 mmol/L) (80.63 (74.62) pg/ml (mean (SD)) compared with 10.25 (9.61) pg/ml.). Moreover, an inverse correlation (r = -0.43, p < 0.05) was found between portal blood IL-6 concentrations and circulating HDL concentrations. CONCLUSIONS: We suggest that serum HDL concentrations influence IL-6 release in response to abdominal operations. Circulating HDL particles may reduce endotoxin-induced IL-6 release in humans. PMID- 11419550 TI - Complete dehiscence of the abdominal wound and incriminating factors. AB - OBJECTIVE: To find out the causes of abdominal wound dehiscence. DESIGN: Retrospective study. SETTING: University hospital, Greece. SUBJECTS: Abdominal wound dehiscence occurred in 89 cases out of 19,206 major abdominal operations including 4671 emergencies during the past 15 years (0.5%). INTERVENTIONS: In the study group 14 local and systemic risk factors were analysed and compared with those in a control group of 89 patients who had similar procedures without dehiscence. MAIN OUTCOME MEASURES: Statistical analysis using the chi square test. RESULTS: Significant factors (p < 0.05) included age over 65 years, emergency operation, cancer, haemodynamic instability, intra-abdominal sepsis, wound infection, hypoalbuminaemia, ascites, obesity, and steroids. Risk factors that were not significant included sex, anaemia, diabetes mellitus and pulmonary disease. Overall morbidity and mortality were 30% and 16%, respectively. The mortality and the possibility of dehiscence seem to correlate directly with the number of risk factors. CONCLUSION: Patients with these risk factors require more attention and special care to minimise the risk of its occurrence. PMID- 11419551 TI - Load on the inguinal region under standard conditions in pigs. AB - OBJECTIVE: To find out the effect of standardised stress on the load of the inguinal region. DESIGN: Experimental study. SETTING: Surgical university clinic, Germany. SUBJECTS: 15 uncastrated adult male pigs. INTERVENTIONS: The tension force between the lateral edge of the rectus sheath and the base of the inguinal ligament was measured after simulation of the Shouldice repair. Standard stresses (coughing, straightening up into the upright position, small or large pneumoperitoneum, and electrical stimulation of abdominal muscles) were imposed. MAIN OUTCOME MEASURES: Intra-abdominal pressure and tension force in the inguinal region. RESULTS: We found the highest (mean, SEM) intra-abdominal pressure during coughing (33.1, 5.2 mm Hg) with an increase in inguinal tension force of 4.1, 0.5 N. The upright position (20, 0.7 mm Hg) and the large pneumoperitoneum (25 mm Hg) produced similar values (4.7, 0.8 N and 3.7, 0.4 N, respectively), whereas the small pneumoperitoneum (10 mm Hg) and the electrical stimulation of muscles of the abdominal wall (4.4, 0.1 mm Hg) resulted in distinctly smaller changes (1.1, 0.2 N and 0.7, 0.1 N, respectively). CONCLUSIONS: The amount of tension on the simulated Shouldice repair depended largely on the increase in intra-abdominal pressure. Compared with the force required to disrupt the normal inguinal canal (68.6 N) the inguinal load seems to be negligibly low. This finding strengthens the modern recommendation to allow only a short convalescence after Shouldice hernia repair. PMID- 11419552 TI - Intragastric endoscopic mucosal resection through a temporary gastrostomy for early gastric cancer: usefulness of Buess-type endoscope. AB - OBJECTIVE: To assess the feasibility of cure of early gastric cancer by the endoscopic procedure done through a temporary gastrostomy and to use it to treat patients with various severe complications. DESIGN: Prospective study. SETTING: University hospital, Japan. SUBJECTS: Four patients who required gastric mucosal resection for early gastric cancer. MAIN OUTCOME MEASURES: Ease of the procedure, cure rate, safety under epidural anaesthesia, and outcome. RESULTS: All patients had the procedure under epidural anaesthesia without pain or complaints during operation. Mean operation time was 120 minutes (60-200). Histological examination showed that the surgical margin was free of tumour in 3 of the cases. One patient who had microscopic invasion of lymph nodes subsequently underwent second-look conventional gastrectomy with lymphadenectomy. Postoperative recovery was rapid, and all patients were discharged from the hospital uneventfully. CONCLUSION: This organ-preserving procedure enables good visualisation of the tumour and is useful even for high-risk patients. PMID- 11419553 TI - Effect of granulocyte-macrophage colony stimulating factor on bacterial translocation after experimental obstructive jaundice. AB - OBJECTIVE: To investigate the effects of granulocyte-macrophage colony stimulating factor (GM-CSF) on bacterial translocation promoted by obstructive jaundice. DESIGN: Controlled animal study. SETTING: University hospital, Turkey. ANIMALS: 30 male Wistar albino rats. INTERVENTIONS: The first group (n = 10) was the sham operation (control) group, and the second and the third (n = 10 each) had common bile duct (CBD) ligation and division under sterile conditions. The third group were also treated with GM-CSF 200 ng subcutaneously daily between the fifth and ninth postoperative days. All animals were killed on the tenth day, and evaluated biochemically and histopathologically. Mesenteric lymph nodes were cultured under aerobic conditions. MAIN OUTCOME MEASURES: Biochemical analysis, histopathological evaluation, and aerobic cultures. RESULTS: There was no bacterial translocation in either the control or GM-CSF groups, whereas Escherichia coli and Salmonella typhimurium were found in 4 and 2 animals, respectively in the ligation group. Although no aerobic bacteria was found in controls and the GM-CSF groups, bacterial translocation was 6/10 in the ligation alone group (p <0.01). CONCLUSION: Activation of inflammatory response with GM CSF is highly effective in prevention of bacterial translocation in obstructive jaundice. PMID- 11419554 TI - Enteric conversion after bladder-drained pancreatic transplantation; a simple and safe salvage procedure. AB - OBJECTIVE: To present our experience with conversion from bladder to enteric drainage after simultaneous pancreatic and renal transplants, so that new transplant centres know that it is both safe and effective. DESIGN: Retrospective study. SETTINGS: Teaching hospital, Republic of Ireland. SUBJECTS: Six patients who had simultaneous pancreatic and renal transplants for insulin-dependent diabetes and who subsequently developed complications of bladder drainage including recurrent episodes of dehydration and metabolic acidosis (n = 3), haematuria (n = 2), and urinary tract infections (n = 1). INTERVENTION: Conversion to enteric drainage. MAIN OUTCOME MEASURE: Resolution of symptoms. RESULTS: All symptoms resolved, but one patient each developed pulmonary oedema, haematuria, and prolonged ileus. All three complications resolved on conservative treatment. All patients are well with surviving grafts a mean of 40 months later (range 19-50). CONCLUSION: Conversion to enteric drainage is safe and effective in patients with refractory metabolic or urological complications of bladder drainage after simultaneous pancreatic and renal transplantation. PMID- 11419555 TI - Sulindac in familial adenomatous polyposis: evaluation by nuclear morphometry. AB - OBJECTIVE: To find out if nuclear morphometry is a useful indicator of prognosis in patients with familial adenomatous polyposis (FAP) before and after treatment with sulindac. DESIGN: Open study. SETTING: Teaching hospital, Spain. SUBJECTS: 29 patients (17 women and 12 men) with FAP who had been treated by colectomy. There were two control groups: 17 people with healthy rectal mucosa and 10 patients with colorectal cancer. INTERVENTIONS: The 29 patients with FAP were treated with sulindac 150 mg orally twice daily for 6 months. MAIN OUTCOME MEASURES: Results of histological examination of biopsy specimens and nuclear morphometry. RESULTS: Before treatment, all patients with FAP had nuclei of cells in the rectal mucosa that were intermediate in size between those in healthy mucosa (small nuclei) and those in cancer (large nuclei). After 6 months' treatment with sulindac the patients with FAP no longer had any polyps and the size of the nuclei was similar to those in healthy mucosa. Only 1 patient failed to respond, and in this patient one of the polyps subsequently became malignant. CONCLUSIONS: Nuclear morphometry may be useful in identifying patients with FAP who are at high risk of malignant degeneration. PMID- 11419556 TI - Isosorbide dinitrate in the treatment of anal fissure: a randomised, prospective, double blind, placebo-controlled trial. AB - OBJECTIVE: To assess the efficacy of isosorbide dinitrate in healing anal fissures. DESIGN: Randomised, prospective, double blind, placebo controlled trial. SETTING: Teaching hospital, The Netherlands. SUBJECTS: 37 consecutive subjects with anal fissure diagnosed in the surgical outpatient department. INTERVENTIONS: After randomisation, 20 patients were given isosorbide dinitrate, and 17 patients placebo. MAIN OUTCOME MEASURES: Healing of anal fissure, recurrence, and tolerance. RESULTS: Both groups were treated for a median (range) of 5 weeks (range 1-10). After this period, 17 in the isosorbide group had healed compared with 6 controls (p < 0.003). The fissure recurred in 2 patients who had had an initial good response to isosorbide, and in 2 in the control group. Side effects (particularly headache) were more common after isosorbide dinitrate, but not significantly so (9/20 compared with 3/17). CONCLUSIONS: Isosorbide dinitrate is an effective treatment for anal fissure, and is significantly better than placebo. PMID- 11419557 TI - Sentinel node biopsy in a patient with two melanomas of the trunk. PMID- 11419558 TI - Disruption of the left gastric artery after apparently minor blunt abdominal trauma. PMID- 11419559 TI - Normalisation of blood pressure and heart rate after excision of a thyroid paraganglioma. PMID- 11419560 TI - Metastases in the amygdala as the first sign of papillary thyroid carcinoma. PMID- 11419561 TI - Evaluation of drug-induced QT interval prolongation: implications for drug approval and labelling. AB - Assessment of proarrhythmic toxicity of newly developed drugs attracts significant attention from drug developers and regulatory agencies. Although no guidelines exist for such assessment, the present experience allows several key suggestions to be made and an appropriate technology to be proposed. Several different in vitro and in vitro preclinical models exist that, in many instances, correctly predict the clinical outcome. However, the correspondence between different preclinical models is not absolute. None of the available models has been demonstrated to be more predictive and/or superior to others. Generally, compounds that do not generate any adverse preclinical signal are less likely to lead to cardiac toxicity in humans. Nevertheless, differences in likelihood offer no guarantee compared with entities with a preclinical signal. Thus, the preclinical investigations lead to probabilistic answers with the possibility of both false positive and false negative findings. Clinical assessment of drug induced QT interval prolongation is crucially dependent on the quality of electrocardiographic data and the appropriateness of electrocardiographic analyses. An integral part of this is a precise heart rate correction of QT interval, which has been shown to require the assessment of QT/RR relationship in each study individual. The numbers of electrocardiograms required for such an assessment are larger than usually obtained in pharmacokinetic studies. Thus, cardiac safety considerations need to be an integral part of early phase I/II studies. Once proarrhythmic safety has been established in phase I/II studies, large phase III studies and postmarketing surveillance can be limited to less strict designs. The incidence of torsade de pointes tachycardia varies from 1 to 5% with clearly proarrhythmic drugs (e.g. quinidine) to 1 in hundreds of thousands with drugs that are still considered unsafe (e.g. terfenadine, cisapride). Thus, not recording any torsade de pointes tachycardia during large phase III studies offers no guarantee, and the clinical premarketing evaluation has to rely on the assessment of QT interval changes. However, since QT interval prolongation is only an indirect surrogate of predisposition to the induction of torsade de pointes tachycardia, any conclusion that a drug is safe should be reserved until postmarketing surveillance data are reviewed. The area of drug related cardiac proarrhythmic toxicity is fast evolving. The academic perspective includes identification of markers more focused compared with simple QT interval measurement, as well as identification of individuals with an increased risk of torsade de pointes. The regulatory perspective includes careful adaptation of new research findings. PMID- 11419562 TI - Management of chemotherapy-induced adverse effects in the treatment of colorectal cancer. AB - The anticancer agents fluorouracil, raltitrexed, irinotecan and oxaliplatin show limited efficacy in the treatment of colorectal cancer and may be associated with substantial toxicity. Therefore, the prevention and reduction of chemotherapy induced adverse effects is of major significance, in accordance with the increasing concern for the quality of life of patients with cancer. Therapeutic drug monitoring of fluorouracil and chronomodulation of fluorouracil and oxaliplatin, have been effective in reducing the incidence and gravity of adverse effects in several clinical trials. However, these concepts have not been implemented in clinical practice yet. At the present time, dose adaptation and supportive measures are the main tools for toxicity control in the treatment of colorectal cancer. In this review, supportive measures for alleviation of the adverse effects of fluorouracil, raltitrexed, irinotecan and oxaliplatin, respectively, are described, based on study results. The main adverse effects of these agents are myelosuppression, oral mucositis, diarrhoea, acute cholinergic syndrome, nausea and emesis, neurotoxicity, hand-foot syndrome and other cutaneous adverse effects, ocular toxicity, cardiotoxicity, small bowel toxicity, asthenia, elevated liver transaminase levels and alopecia. The incidence and gravity of these adverse effects are more or less related to the agent and administration schedule involved. The supportive measures and recommendations include the use of specific drugs, alterations of administration schedule and several nonpharmacological methods. In addition, guidelines for dosage adjustments when toxicity occurs are presented. For optimal management of adverse effects, patients should be considered individually, while patients, nurses and physicians should cooperate to identify and treat adverse effects in an early stage of their development. PMID- 11419563 TI - Skin reactions to inhaled corticosteroids. AB - Corticosteroids intended for inhalation into the lungs or into the nose have been used since the 1970s. Only 2 attempts to assess contact allergy attributable to inhaled corticosteroids in patients with asthma and/or rhinitis have been made, and only 1 single case of contact allergy attributable to budesonide and tixocortol pivalate was found. However, several case reports of allergic mucosal and skin symptoms caused by corticosteroids applied locally to the mucosa have been published. Local adverse effects from nasal corticosteroids have ranged from nasal congestion, pruritus, burning, and soreness to perforation of the nasal septum. Inhalation of corticosteroids into the lungs has been reported to cause pruritus, dryness, erythema and oedema of the mouth, a dry cough and odynophagia. Systemic signs reported from the use of nasal corticosteroids and inhalation of corticosteroids into the lungs have been eczematous lesions, particularly on the face, sometimes with spreading to the trunk and flexures. Urticaria has also been noted. PMID- 11419564 TI - Tolerability of treatments for viral hepatitis. AB - Interferon-alpha is the most widely used antiviral drug in chronic hepatitis B and C. Tolerability is usually good and serious adverse effects are rare. Most of the adverse effects are mild or transient and do not necessitate drug withdrawal. More than 90% of patients who are given interferon-alpha achieve 6 months to 1 year of treatment without serious adverse effects. The serious adverse effects usually occur in predisposed patients with pre-existing organ dysfunction. Nevertheless, careful selection of patients for therapy and observation during therapy are recommended. Nucleoside analogues are promising drugs in the treatment of chronic hepatitis B through inhibition of viral DNA polymerase. Lamivudine has been licensed for use in this indication. Its tolerability is excellent even when used for periods of 1 year or more. The main concern is the relatively high incidence of viral resistance resulting in breakthrough during or relapse after therapy. In the treatment of chronic hepatitis C, ribavirin, in combination with interferon-alpha is currently the reference therapy. The main adverse effect is haemolytic anaemia, which necessitates careful monitoring and adjustment of dosage in many cases. Recently, large trials showed the better efficacy of pegylated interferons as compared with standard interferon. The combination of pegylated interferon with ribavirin is under evaluation. PMID- 11419565 TI - Visual field defects and other ophthalmological disturbances associated with vigabatrin. AB - Vigabatrin has been an important anticonvulsant drug for over 10 years with a reputation for high efficacy and excellent tolerability. However, since 1997, there have been over 25 reports in the literature of visual field defects attributable to the use of this agent. Most are case reports and many have only been reported as abstracts or posters or as letters or short communications. Only a small number of papers give details of patient characteristics. Typically, case reports detail ophthalmological tests such as visual acuity, funduscopic examination, integrity of colour vision, and the nature of the field cut. Many also include various electrophysiological tests which were performed in an attempt to further describe the nature of the visual changes. In order to shed light on the mechanism underlying these visual field changes, many investigators also tested various electrophysiological parameters. However, because electrophysiological testing requires considerable expertise on the part of the technician, this could be a source of variability in results and may also pose a challenge with data interpretation. The magnitude of the problem is difficult to assess. The manufacturer's estimate of incidence of visual field defects with vigabatrin was approximately 0.1%, but incidences estimated in the literature range from 6 to 30%. Since the majority of the published data are in case report form, proof of causation is also very difficult. Two papers that used proper scientific methodology to investigate this condition suggest that vigabatrin causes these changes; however, there needs to be further studies with larger populations to answer this question definitively. There is a lack of data on the dose-response characteristics of vigabatrin and the development of visual field defects. The only available data are reports of trends that implicate duration of therapy or cumulative dose. Perhaps the most important area to elucidate is whether or not the visual field defects are reversible. Data are scare on this subject, but we can hope that data will emerge as follow-up periods become more substantial. There is a need for more complete information regarding several aspects of the mechanistic basis of visual field defects associated with vigabatrin that will allow rational clinical decision making. The treatment choices, both pharmacological and nonpharmacological, for patients with refractory epilepsy have grown substantially in the last few years. Thus, it is doubtful that the clinical positioning of vigabatrin is likely to change in the future from that of a very valuable 'niche drug', with emphasis on paediatric usage. PMID- 11419566 TI - Efficacy of metrifonate in improving the psychiatric and behavioral disturbances of patients with Alzheimer's disease. AB - Neuropsychiatric and behavioral symptoms are frequent and problematic components of Alzheimer's disease (AD). In two previously reported studies, metrifonate was shown to benefit behavioral symptoms as assessed by the Neuropsychiatric Inventory (NPI). In this post hoc analysis, detailed studies were completed to determine the effects of metrifonate on individual symptoms. This study was a retrospective analysis of pooled NPI data from two double-blind, placebo controlled, multicenter 26-week studies of metrifonate that had achieved similar levels of cholinesterase inhibition. Mild-to-moderate probable AD patients received placebo (n = 222) or metrifonate (n = 450) 30 to 60 mg by weight or a 50 mg fixed dose once daily. At 26 weeks, metrifonate-treated patients had significantly reduced NPI total scores (P = .001) and fewer neuropsychiatric symptoms when compared with placebo-treated patients, including hallucinations (P = .004), agitation/aggression (P = .006), depression/dysphoria (P = .011), apathy (P = .019), and aberrant motor behavior (P = .008). Metrifonate reduced or stabilized neuropsychiatric disturbances in 60% of symptomatic patients. Almost 40% of metrifonate-treated patients had a clinically relevant reduction (> or = 30% decrease in NPI score) in their neuropsychiatric disturbances (P = .002). High proportions of metrifonate-treated patients manifested clinically relevant reductions in anxiety (58%, P = .009), apathy (51%, P = .020), and depression/dysphoria (50%, P = .021) compared to placebo. The metrifonate associated reductions in NPI scores were evident by week 12 and were maintained for the 26-week study period. There was an overall effect size of metrifonate of approximately 15% on total NPI scores when compared to placebo. Metrifonate significantly reduced many of the psychiatric and behavioral symptoms of AD. The observations suggest that enhancement of cholinergic functions in AD has beneficial effects on behavior. PMID- 11419567 TI - Telemedicine and geriatric psychiatry: directions for future research and policy. AB - The use of telecommunications to provide mental health services at a distance has grown rapidly in the past 10 years. The overall experience has been positive, but evaluations and reliability studies have been preliminary. A fundamental question of what constitutes adequate technology for telepsychiatry remains unanswered. As technology and equipment capability change rapidly, a more important question may be what clinical decisions and behavioral observations are required to produce high standards of distance mental health care. Geriatric patients may have sensory impairments and unique aspects to their psychiatric problems that can make telemental health assessments more challenging. It is not clear what model of telepsychiatry constitutes the best practice for geriatric psychiatry. Future research and program evaluations should address these questions to guide the use of telepsychiatry in productive directions. PMID- 11419568 TI - Psychiatric assessment of a nursing home population using audiovisual telecommunication. AB - The purpose of this study was to demonstrate that psychiatric assessment of nursing home residents could be reliably carried out remotely via telecommunications. Twenty-seven nursing home residents each had two interviews consisting of the following three rating scales: the Mini-Mental State Examination (MMSE), the Geriatric Depression Scale (GDS), and the Brief Psychiatric Rating Scale (BPRS). The interviews were conducted by three trained psychiatrists, each of whom interviewed two-thirds of the subjects. Subjects were sequentially assigned to have either two in-person interviews (in-person group) or one in-person and one remote interview via telecommunication (remote group). Inter-rater reliability was calculated separately for each condition (in-person vs remote group) for each of the three rating scales. Intraclass correlations on the MMSE were .95 for the remote group and .83 for the in-person group. On the GDS, they were .82 for the remote group and .86 for the in-person group. Finally, on the BPRS, they were .81 for the remote group and .49 for the in-person group. There were no statistically significant differences in intraclass correlation on any of the three scales for the remote group compared with the in-person group, indicating that nursing home residents can be reliably assessed remotely via telecommunication. PMID- 11419569 TI - Reliability of telepsychiatry assessments: subjective versus observational ratings. AB - Geriatric patients in underserved areas could benefit from the use of telecommunications to expand access to mental health services. It is important to determine the clinical limitations of using videoconferencing for psychiatric assessments, particularly in the elderly. The goal of this study was to test the hypothesis that videoconferencing ratings based on visual observations of behavior would be less reliable than ratings based on patients' verbal reports of symptoms. Videoconferencing assessments of 30 geriatric patients using low bandwidth (ISDN) equipment were compared to gold standard face-to-face assessments. The Brief Psychiatric Rating Scale (BPRS) was dichotomized into subjective items based on patients' verbal reports and observational items based on visual ratings of behavior. Reliability of the BPRS subjective items was consistently higher than for the observational items. Future studies should emphasize the accuracy of telemedicine ratings that require visual observation of behavior, which is crucial to clinical assessment of psychogeriatric conditions. PMID- 11419570 TI - Telepsychiatry consultations to a rural nursing facility: a 2-year experience. AB - This report describes the operation of a telepsychiatry consultation service provided from an academic medical center to a skilled nursing facility in a rural setting. Seventy-one consultations via low-cost ISDN videoconferencing equipment were provided over the 2-year period to 40 patients. The average duration of the consultation period was about 3 months. A wide variety of diagnostic conditions were encountered, with the majority being either depression or dementia-related behavior problems. Advantages of the program included the ability to respond promptly to rural patients' clinical needs, the flexibility to consult more frequently if clinically indicated, and more efficient use of the consultant's time, as travel time was eliminated. The program is an example of how telepsychiatry can successfully expand access to mental health services to residents of long-term care facilities. PMID- 11419571 TI - Telemedicine use and the reduction of psychiatric admissions from a long-term care facility. AB - Copper Ridge is a long-term care facility that provides care for persons with dementia and their families from early diagnosis to end of life. A low-cost videoconferencing system was employed in the development of a comprehensive, integrated continuum of care for Copper Ridge residents by bridging long-term care with inpatient psychiatric care at Johns Hopkins Hospital. In this article, we discuss the Copper Ridge/Johns Hopkins telemedicine project and how its operation appears to have brought about a reduction in psychiatric admissions. Telemedicine projects using inexpensive technology over standard telephone lines can be successfully used in long-term care settings. PMID- 11419572 TI - Program management of telemental health care services. AB - Telemedicine is a new adjunct to the delivery of health care services that has been applied to a range of health care specialties, including mental health. When prospective telemedicine programs are planned, telemedicine is often envisaged as simply a question of introducing new technology. The development of a robust, sustainable telemental health program involves clinical, technical, and managerial considerations. The major barriers to making this happen are usually how practitioners and patients adapt successfully to the technology and not in the physical installation of telecommunications bandwidth and the associated hardware necessary for teleconsultation. This article outlines the requirements for establishing a viable telemental health service, one that is based on clinical need, practitioner acceptance, technical reliability, and revenue generation. It concludes that the major challenge associated with the implementation of telemental health does not lie in having the idea or in taking the idea to the project stage needed for proof of concept. The major challenge to the widespread adoption of telemental health is paying sufficient attention to the myriad of details needed to integrate models of remote health care delivery into the wider health care system. PMID- 11419573 TI - Geriatric telepsychiatry: no matter the population, the questions remain the same -a commentary. PMID- 11419574 TI - Caregivers' judgments of the functional abilities of the Alzheimer's disease patient: impact of caregivers' depression and perceived burden. AB - Research has demonstrated that caregivers of cognitively impaired patients may misjudge aspects of the patient's functional capacities. The nature and directions of these relationships are not well understood, however. Further, the effects that depression and perceived caregiver burden have on the caregivers' ability to render accurate judgments of patient's functional abilities have not been addressed. In this study, the primary caregivers of 128 patients with Alzheimer's disease (AD) were administered a questionnaire regarding the patients' functional capacities. These judgments were subsequently compared to actual functional performance of AD patients on an extended version of the Direct Assessment of Functional Status scale administered in the patients' home environment. A significant proportion of caregivers overestimated AD patients' functional performance in telling time, counting currency, making change for a purchase, brushing teeth, and using eating utensils. Further analyses revealed that self-reported depression, but not perceived burden, was related to the type of caregiver bias errors observed. PMID- 11419575 TI - Quetiapine-Induced extrapyramidal side effects in patients with Parkinson's disease: case report. AB - Although quetiapine is the antipsychotic of choice for the psychosis associated with Parkinson's disease (PD) and often is also helpful for sleep, we report two cases of quetiapine-induced extrapyramidal side effects. The patients described were unusual in their frailty and severity of illness and may not represent the majority of patients with PD. PMID- 11419577 TI - Making the case for health interventions in correctional facilities. PMID- 11419576 TI - Jails, prisons, and the health of urban populations: a review of the impact of the correctional system on community health. AB - This review examined the interactions between the correctional system and the health of urban populations. Cities have more poor people, more people of color, and higher crime rates than suburban and rural areas; thus, urban populations are overrepresented in the nation's jails and prisons. As a result, US incarceration policies and programs have a disproportionate impact on urban communities, especially black and Latino ones. Health conditions that are overrepresented in incarcerated populations include substance abuse, human immunodeficiency virus (HIV) and other infectious diseases, perpetration and victimization by violence, mental illness, chronic disease, and reproductive health problems. Correctional systems have direct and indirect effects on health. Indirectly, they influence family structure, economic opportunities, political participation, and normative community values on sex, drugs, and violence. Current correctional policies also divert resources from other social needs. Correctional systems can have a direct effect on the health of urban populations by offering health care and health promotion in jails and prisons, by linking inmates to community services after release, and by assisting in the process of community reintegration. Specific recommendations for action and research to reduce the adverse health and social consequences of current incarceration policies are offered. PMID- 11419578 TI - Characteristics and trends of newly identified HIV infections among incarcerated populations: CDC HIV voluntary counseling, testing, and referral system, 1992 1998. AB - Inmate contact with the correctional health care system provides public health professionals an opportunity to offer HIV screening to a population that might prove difficult to reach otherwise. We report on publicly funded human immunodeficiency virus (HIV) voluntary counseling, testing, and referral (VCTR) services provided to incarcerated persons in the United States. Incarcerated persons seeking VCTR services received pretest counseling and gave a blood specimen for HIV antibody testing. Specimens were considered positive if the enzyme immunoassays were repeatedly reactive and the Western blot or immunofluorescent assay was reactive. Demographics, HIV risk information, and laboratory test results were collected from each test episode. Additional counseling sessions provided more data. From 1992 to 1998, there were 527,937 records available from correctional facilities from 48 project areas; 484,277 records included a test result and 459,155 (87.0%) tests came with complete data. Overall, 3.4% (16,797) of all tests were reactive for HIV antibodies. Of reactive tests accompanied by self-reports of previous HIV test results (15,888), previous test results were 44% positive, 23% negative, 6% inconclusive or unspecified, and 27% no previous test. This indicates that 56% of positive tests were newly identified. During the study period, the number of tests per year increased three fold. Testing increased among all racial/ethnic groups and both sexes. The largest increase was for heterosexuals who reported no other risk, followed by persons with a sex partner at risk. Overall, the greatest number of tests was reported for injection drug users (IDUs) (128,262), followed by men who have sex with men (MSM) (19,928); however, episodes for MSM doubled during the study, while for IDUs, they increased 74%. The absolute number of HIV-positive (HIV+) tests increased 50%; however, the percentage of all tests that were HIV+ decreased nearly 50% due to the increased number of tests performed. HIV+ tests fell 50% among blacks (7.6% to 3.7%), Hispanics (6.7% to 2.5%), and males (5.1% to 2.5%); 33% among females (4.5% to 3.1%); 95% among IDUs (8.6% to 4.4%); and 64% among MSM (19.3% to 11.8%). Among HIV+ episodes, those for IDUs dropped from 61.5% to 36.6%, while episodes for heterosexuals with no reported risk factor increased from 4.3% to 18.2%. The use of VCTR services by incarcerated persons rose steadily from 1992 to 1998, and 56% of HIV+ tests were newly identified. High numbers of tests that recorded risk behaviors for contracting HIV indicate that correctional facilities provide an important access point for prevention efforts. PMID- 11419579 TI - HIV and AIDS surveillance among inmates in Maryland prisons. AB - The prevalence of those with human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS) is higher among inmates of correctional facilities than among the general population. This raises the need to identify inmates living with or at risk of HIV/AIDS and to provide counseling and appropriate services for HIV treatment and prevention. The Maryland Division of Corrections (DOC) offers voluntary testing to all inmates on entry and tests inmates when clinically indicated. We reviewed all 1998 HIV antibody tests and confirmed AIDS cases in the Maryland DOC. Inmate demographics, testing acceptance, rates of seropositivity, and AIDS cases and comparisons based on gender, race/ethnicity, and age were examined. Comparisons were also made to HIV testing and AIDS cases from the nonincarcerated Maryland population. Trends in DOC AIDS diagnoses and AIDS-related deaths over time were also examined. Of the inmates, 39% were voluntarily tested for HIV on entry to the Maryland DOC in 1998 (38% of males and 49% of females). Overall, HIV seropositivity was 3.3% (5% for females and 3% for males). The 888 cumulative AIDS cases diagnosed in the DOC inmate population were concentrated among males (90% vs. 77% statewide), African Americans (91% vs. 75% statewide), and among IDUs (84% vs. 39% statewide). Due to high rates of HIV and AIDS, inmate populations are a crucial audience for HIV/AIDS testing, treatment, and prevention efforts, especially women. Prison based programs can identify significant numbers of HIV and AIDS cases and bring HIV prevention interventions to a population characterized by frequent high-risk behavior. PMID- 11419580 TI - Using a jail-based survey to monitor HIV and risk behaviors among Seattle area injection drug users. AB - Routine monitoring of human immunodeficiency virus (HIV) and risk behaviors among injection drug users (IDUs) is difficult outside drug treatment settings. We developed and implemented a survey of recently arrested IDUs to describe the prevalence of HIV, drug use, and sexual behaviors among them. A probability sampling survey was instituted in the King County Correctional Facility in Seattle, Washington, to sample recently arrested IDUs at the time of booking and in the jail health clinic between 1998 and 1999. Following HIV risk assessment and blood draw, additional information on drug use practices was gathered using a standardized questionnaire. Potential participants who were released from jail early could complete the study at a nearby research storefront office. Of the 4,344 persons intercepted at booking, 503 (12%) reported injection drug use, and 201 of the IDUs (40%) participated in the study. An additional 161 IDUs were enrolled in the study from the jail health clinic. Among the 348 unduplicated subjects, HIV prevalence was 2%; in the past 6 months, 69% reported two or more shooting partners, 72% used a cooker after someone else, 60% shared a syringe to divide up drugs, and 62% injected with used needles. Only 37% reported being hepatitis C seropositive, and 8% reported hepatitis B vaccination. It was feasible to conduct a jail-based survey of recently arrested IDUs that yielded useful information. The high prevalence of reported risky drug use practices warrants ongoing monitoring and illustrates the need for improving prevention programs for HIV and hepatitis B and C in this population, including expansion of hepatitis C screening and provision of hepatitis B vaccination at the jail health clinic. PMID- 11419581 TI - Successful linkage of medical care and community services for HIV-positive offenders being released from prison. AB - Human immunodeficiency virus (HIV) infection is more prevalent among the incarcerated than the general population. For many offenders, incarceration is the only time that they may access primary care. Project Bridge is a federally funded demonstration project that provides intensive case management for HIV positive ex-offenders being released from the Rhode Island state prison to the community. The program is based on collaboration between colocated medical and social work staff. The primary goal of the program is to increase continuity of medical care through social stabilization; it follows a harm reduction philosophy in addressing substance use. Program participants are provided with assistance in accessing a variety of medical and social services. The treatment plan may include the following: mental illness triage and referral, substance abuse assessment and treatment, appointments for HIV and other medical conditions, and referral for assistance to community programs that address basic survival needs. In the first 3 years of this program, 97 offenders were enrolled. Injection drug use was reported by 80% of those enrolled. There were 90% followed for 18 months, 7% moved out of state or died, and 3% were lost to follow-up. Reincarceration happened to 48% at least once. Of those expressing a need, 75% were linked with specialty medical care in the community, and 100% received HIV-related medical services. Of those expressing a need for substance abuse treatment, 67% were successful in keeping appointments for substance abuse treatment within the community. Project Bridge has demonstrated that it is possible to maintain HIV positive ex-offenders in medical care through the provision of ongoing case management services following prison release. Ex-offenders will access HIV related health care after release when given adequate support. PMID- 11419582 TI - Reintegrating women leaving jail into urban communities: a description of a model program. AB - Women are the fastest-growing population in the criminal justice system, and jails reach more people than any other component of the correctional system. About 1 million women pass through US jails each year. Most return to their communities within a few weeks of arrest, and few receive help for the substance abuse, health, psychological or social problems that contribute to incarceration. We describe a model program, Health Link, designed to assist drug-using jailed women in New York City to return to their communities, reduce drug use and HIV risk behavior, and avoid rearrest. The program operates on four levels: direct services, including case management for individual women in the jail and for 1 year after release; technical assistance, training, and financial support for community service providers that serve ex-offenders; staff support for a network of local service providers that coordinate services and advocate for resources; and policy analysis and advocacy to identify and reduce barriers to successful community reintegration of women released from jail. We describe the characteristics of 386 women enrolled in Health Link in 1997 and 1998; define the elements of this intervention; and assess the lessons we have learned from 10 years of experience working with jailed women. PMID- 11419583 TI - Cost-effectiveness of HIV counseling and testing in US prisons. AB - The prevalence of human immunodeficiency virus (HIV) in correctional facilities is much higher than in the general population. However, HIV prevention resources are limited, making it important to evaluate different prevention programs in prison settings. Our study presents the cost-effectiveness of offering HIV counseling and testing (CT) to soon-to-be-released inmates in US prisons. A decision model was used to estimate the costs and benefits (averted HIV cases) of HIV testing and counseling compared to no CT from a societal perspective. Model parameters were HIV prevalence among otherwise untested inmates (1%); acceptance of CT (50%); risk for HIV transmission from infected individuals (7%); risk of HIV acquisition for uninfected individuals (0.3%); and reduction of risk after counseling for those infected (25%) and uninfected (20%). Marginal costs of testing and counseling per person were used (no fixed costs). If infected, the cost was $78.17; if uninfected, it was $24.63. A lifetime treatment cost of $186,900 was used to estimate the benefits of prevented HIV infections. Sensitivity and threshold analysis were done to test the robustness of these parameters. Our baseline model shows that, compared to no CT, offering CT to 10,000 inmates detects 50 new or previously undiagnosed infections and averts 4 future cases of HIV at a cost of $125,000 to prison systems. However, this will save society over $550,000. Increase in HIV prevalence, risk of transmission, or effectiveness of counseling increased societal savings. As prevalence increases, focusing on HIV-infected inmates prevents additional future infections; however, when HIV prevalence is less than 5%, testing and counseling of both infected and uninfected inmates are important for HIV prevention. PMID- 11419584 TI - Needle-exchange participation, effectiveness, and policy: syringe relay, gender, and the paradox of public health. AB - Needle-exchange programs (NEPs) have been politically controversial, and most studies have focused on evaluating their effectiveness on human immunodeficiency virus (HIV) transmission rates with little emphasis on the process of how they are used. This article shows that the way intravenous drug users use NEPs may influence their effectiveness. Using data from Baltimore's NEP, participants (N = 2,574) were classified as low, medium, and high users based on the volume, frequency, and duration of contact with the NEP. Higher NEP use was associated with shorter syringe circulation times and less syringe relay, returning syringes to the NEP originally acquired by someone else. For a subsample that was HIV tested (N = 262), syringe relay among women was associated with HIV seroconversion (at a 95% confidence interval). We conclude that exclusive use of the NEP (no relay) provides greater HIV protection than NEP use involving syringe relay. The paradox is that public health goals will not be achieved by prohibiting syringe relay activities and promoting exclusive use. NEPs should broaden their education efforts to have participants understand the value of repeated visits to the NEP. PMID- 11419585 TI - The challenges of emerging illness in urban environments: an overview. AB - The New York City West Nile outbreak is an important case study for examining several medical and public health issues raised by the specter of emerging illnesses in urban areas. Five specific issues are addressed in this issue of the Journal of Urban Health: ecosystem health, vector (e.g., mosquito) control, risk communication, public health infrastructure, and parallels between this outbreak and bioterrorism. PMID- 11419586 TI - The control of mosquito-borne diseases in New York City. AB - Mosquito control began in New York City in 1901. Large-scale efforts to drain marshlands occurred through the 1930s, and aerial application of pesticide occurred as early as 1956. Components of early mosquito-borne disease control were reimplemented in 1999-2000 in response to an outbreak of West Nile virus, and included promoting public and health professional awareness regarding disease causation and prevention, providing free government laboratory testing, case reporting, mapping of mosquito breeding sites and their elimination or application of larvicide to them, and adult mosquito control. Because a potential for various mosquito-borne diseases in New York City persists, continued efforts are warranted to limit mosquito breeding, to monitor adult mosquito populations for the presence of human pathogens, and to establish protocols and capacity for adult mosquito control. PMID- 11419587 TI - West Nile virus and the climate. AB - West Nile virus is transmitted by urban-dwelling mosquitoes to birds and other animals, with occasional "spillover" to humans. While the means by which West Nile virus was introduced into the Americas in 1999 remain unknown, the climatic conditions that amplify diseases that cycle among urban mosquitoes, birds, and humans are warm winters and spring droughts. This information can be useful in generating early warning systems and mobilizing timely and the most environmentally friendly public health interventions. The extreme weather conditions accompanying long-term climate change may also be contributing to the spread of West Nile virus in the United States and Europe. PMID- 11419588 TI - Balancing the risks: vector control and pesticide use in response to emerging illness. AB - The competing public health concerns of vector-borne disease and vector control strategies, particularly pesticide use, are inherently subjective and difficult to balance. Disease response decisions must frequently be made in the absence of data or clear criteria. The factors to be weighed include the vector control measures versus those posed by the disease itself; short-term versus long-term disease management goals, specifically with regard to the issue of pesticide resistance; the need to distinguish among diseases of differing severity in making response choices; and the issue of pesticide efficacy. New York City's experience with West Nile virus has illustrated each of these issues. A framework for assessing the appropriate response to West Nile virus can serve to guide our response to likely new pathogens. PMID- 11419589 TI - Risk communication, the West Nile virus epidemic, and bioterrorism: responding to the communication challenges posed by the intentional or unintentional release of a pathogen in an urban setting. AB - The intentional or unintentional introduction of a pathogen in an urban setting presents severe communication challenges. Risk communication--a science-based approach for communicating effectively in high-concern situations--provides a set of principles and tools for meeting those challenges. A brief overview of the risk communication theoretical perspective and basic risk communication models is presented here, and the risk communication perspective is applied to the West Nile virus epidemic in New York City in 1999 and 2000 and to a possible bioterrorist event. The purpose is to provide practical information on how perceptions of the risks associated with a disease outbreak might be perceived and how communications would be best managed. PMID- 11419590 TI - West Nile virus: success of public health response underlines failure of the system. AB - The West Nile virus outbreak in 1999 demonstrated the country's capacity to meet an emerging public health threat. However, while the tracking and monitoring efforts that were put into place by 2000 were impressive, the response to the West Nile virus underscores a fundamental deficiency in the capacity of public health regarding the nation's environmental health efforts. Chronic diseases such as asthma, neurological diseases, and birth defects and their potential links to environmental factors are not being adequately tracked and monitored. New public health infrastructure resources are required. PMID- 11419591 TI - Emerging illness and bioterrorism: implications for public health. AB - Biological weapons have the potential to inflict deliberate, potentially devastating epidemics of infectious disease on populations. The science and technology exist to create deliberate outbreaks of human disease, as well as disease among plants and animals, crops, and livestock. A new awareness among policymakers of the link between public health and national security requires the attention of public health professionals. The issues posed by biological weapons are likely to challenge the political assumptions of many progressive public health professionals and will demand new coalitions. The prospect of bioterrorism may offer new opportunities for improving the public health infrastructure and its capabilities. PMID- 11419592 TI - The collapse of global public health and why it matters for New York. PMID- 11419593 TI - Effects of electromagnetic fields produced by radiotelevision broadcasting stations on the immune system of women. AB - The object of this study was to investigate the immune system of 19 women with a mean age of 35 years, for at least 2 years (mean = 13 years) exposed to electromagnetic fields (ELMFs) induced by radiotelevision broadcasting stations in their residential area. In September 1999, the ELMFs (with range 500 KHz-3 GHz) in the balconies of the homes of the women were (mean +/- S.D.) 4.3 +/- 1.4 V/m. Forty-seven women of similar age, smoking habits and atopy composed the control group, with a nearby resident ELMF exposure of < 1.8 V/m. Blood lead and urinary trans-trans muconic acid (a metabolite of benzene), markers of exposure to urban traffic, were higher in the control women. The ELMF exposed group showed a statistically significant reduction of blood NK CD16+-CD56+, cytotoxic CD3(-) CD8+, B and NK activated CD3(-)-HLA-DR+ and CD3(-)-CD25+ lymphocytes. 'In vitro' production of IL-2 and interferon-gamma (INF-gamma) by peripheral blood mononuclear cells (PBMC) of the ELMF exposed group, incubated either with or without phytohaemoagglutinin (PHA), was significantly lower; the 'in vitro' production of IL-2 was significantly correlated with blood CD16+-CD56+ lymphocytes. The stimulation index (S.I.) of blastogenesis (ratio between cell proliferation with and without PHA) of PBMC of ELMF exposed women was lower than that of the control subjects. The S.I. of blastogenesis of the ELMF exposed group (but not blood NK lymphocytes and the 'in vitro' production of IL-2 and INF-gamma by PBMC) was significantly correlated with the ELMF levels. Blood lead and urinary trans-trans muconic acid were barely correlated with immune parameters: the urinary metabolite of benzene of the control group was only correlated with CD16+-CD56+ cells indicating a slight effect of traffic on the immune system. In conclusion, this study demonstrates that high frequency ELMFs reduce cytotoxic activity in the peripheral blood of women without a dose-response effect. PMID- 11419594 TI - Evidence of pre-industrial air pollution from the Heads of the Kings of Juda statues from Notre Dame Cathedral in Paris. AB - Pollution originating from wood combustion characterised the urban atmospheres of the past and led to the formation of thin grey crusts on the surface of the stone of monuments. The grey crusts discovered on the Heads of the Kings of Juda statues, which adorned the facade of Notre Dame in Paris from the 13th century until 1792, constitute a material record of the effects of this ancient air pollution. The height at which the statues stood suggests that the effect was not the result of a point phenomenon, but was caused by a generalised pollution of the Paris atmosphere at the time. PMID- 11419595 TI - Long-term variation (1986-1998) of post-Chernobyl 90Sr, 137Cs, 238Pu and (239,240)Pu concentrations in air, depositions to ground, resuspension factors and resuspension rates in south Germany. AB - Annual mean concentrations in air and annual total (wet plus dry) depositions to ground of 90Sr, 137Cs, 238Pu and (239,240)Pu decreased at Neuherberg, south Germany, in the period from July 1986 (i.e. after the end of the initial deposition phase from Chernobyl) to 1998 as follows: 90Sr from 0.77 to 0.05 microBq m(-3), and from 0.82 (1988) to 0.28 Bq m(-2) year(-1); 137Cs from 133 to 2.8 microBq m(-3), and from 116 to 3.8 Bq m(-2) year(-1); 238Pu from 0.95 to 0.063 nBq m(-3), and from 1.0 to 0.23 mBq m(-2) year(-1); (239,240)Pu from 8.1 to 0.53 nBq m(-3), and from 6.4 to 2.1 mBq m(-2) year(-1). The values for the non caesium radionuclides are compared to the few available data from other stations. After an initial phase which is characterised for a given radionuclide by the varying ratio of the Chernobyl-derived inventory to the earlier, weapons fallout derived inventory, the time courses of concentrations of 137Cs, 90Sr, 238Pu and (239,240)Pu in air become more or less parallel (from about 1990), despite the considerable differences in the physico-chemical properties of these elements. By contrast, the time series of radionuclide deposition rates show until about 1994 less similarity among each other and with the respective concentration series in air. From 1994, concentrations in air and depositions to ground become nearly constant, or decrease very slowly. At the end of the observation period, resuspension factors between 1.4 x 10(-10) m(-1) and 1.0 x 10(-11) m(-1) are observed for the various nuclides. The time courses of specific activities (Bq g( 1)) as well as the time courses of radionuclide ratios show characteristic differences between air and deposition. The data will be useful in predicting the post-accident behaviour of radionuclides a long time after a large-scale contamination event. PMID- 11419596 TI - Geochemical investigations of sulfide-bearing tailings at Kristineberg, northern Sweden, a few years after remediation. AB - In the Kristineberg mining area in northern Sweden, massive, pyrite-rich Zn Cu ores are intercalated in ca. 1.9 Ga volcano-sedimentary rocks. Investigations of a tailings impoundment remediated by means of both till coverage and raising the groundwater table have been undertaken. The aim of the study was to characterise the tailings with respect to mineralogy, the chemical composition of both the tailings and the pore water, and to try to identify the significant reactions that may have occurred before and after remediation. It was found that the oxidation front had reached down to depths of between approximately 0.1 and 1.15 m before remediation. The oxidation of sulfides has produced high concentrations of some metals in the pore water; up to 26, 16, 4.1, 2.7 and 82 mg/l have been measured for Al, Mn, Fe and Zn, respectively. Concentrations of metals such as Cd, Co, Cu, Ni and Pb are lower, with average concentrations of 18.4, 83.8, 45, 79.6 and 451 microg/l, respectively. Higher concentrations of major elements such as Ca, Fe, Mn, Mg and S have been measured at depth in pore water than at shallower levels. This is probably caused by flush out of elements after remediation and vertical transport from the upper parts before remediation. The pH is relatively high, approximately 5.5 at most depths in the tailings, except in and around the former oxidation zone where it is lower, and where the highest dissolved concentrations of elements such as As, Cd, Co, Cu, Pb and Zn occur. This is probably due to the release of metals secondarily retained below the oxidation front prior to the remediation. Since the groundwater table is raised, the groundwater reaches the retained metals, which leads to desorption of metals and dissolution of secondary minerals. PMID- 11419597 TI - Environmental exposure of the pediatric age groups in Cairo City and its suburbs to cadmium pollution. AB - In a trial to assess the exposure of subjects in the pediatric age group to cadmium (Cd) pollution, serum Cd was estimated by atomic absorption spectrometry in 405 subjects, birth-18 years old, from Cairo City and its suburbs. Serum Cd mean concentrations were: 0.92 microg/l in 32 neonates (birth-4 weeks); 1.33 microg/l in 70 infants (4 weeks-2 years); 1.11 microg/l in 100 children in the preschool period (2-6 years); 1.34 microg/l in 103 primary school children (6-12 years); and 1.24 microg/l in 100 adolescents (12-18 years). In neonates, serum Cd was higher in babies with weights and heights that remained below the 5th percentile for age. Breast-fed infants had a serum Cd geometric mean level (1.25 microg/l) that was not in accordance to that of their mothers' milk (0.52 microg/l, P < 0.001), suggesting alternative routes of exposure. Environmental tobacco-smoke exposure was the most important determinant of Cd status in the school-aged children, the geometric mean being 1.42 microg/l in passive smokers vs. 1.2 microg/l in non-exposed children (P < 0.05). Moreover, adolescents who were active smokers had a significantly higher serum Cd level (1.7 microg/l) as compared to non-smokers (1.2 microg/l). Gender did influence the Cd status in adolescents, being higher among males, probably related to smoking, or to the difference in lifestyle of adolescents according to gender in the community. Alpha-1-microglobulinuria was accompanied by a higher serum Cd concentration in the group of adolescents only, suggesting a subclinical renal effect after several years of cumulative exposure. The residential classification, whether urban or suburban, did not influence the serum Cd status; neither did the present or past history of bronchial asthma. These findings certainly justify further evaluation of the problem of Cd pollution among Cairene individuals, knowing the long-term consequences of exposure to it. Systematic efforts for the proper disposal of Cd wastes and prevention of smoking in public places are recommended. PMID- 11419598 TI - Nutrient fluxes in the Po basin. AB - The nutrient load of the river Po needs to be reduced in order to bring the eutrophication problems in the Northwest Adriatic Sea under lasting control. In this paper we analyse the contribution of the different pollution sources to the nitrogen and phosphorus loads of the river Po (source apportionment). We also estimate the effects of measures that aim to reduce nitrogen and phosphorus pollution in the Po basin (scenario analyses). Using a model (PolFlow) that was previously applied to the Rhine and Elbe basins, we found that more than half of the nitrogen and phosphorus loads in the river Po originates from diffuse sources. The results of the scenario analyses indicate that the measures imposed by the EU Nitrates Directive and the EU Wastewater Treatment Directive may not be stringent enough to achieve a large reduction in the N and P loads in the river Po. PMID- 11419599 TI - Daily intakes of 238U, 234U, 232Th, 230Th, 228Th and 226Ra in the adult population of central Poland. AB - Activity concentration of the uranium and thorium series radionuclides was determined in foodstuffs and drinking water in central Poland. Annual and daily intake for the adult population was estimated from the concentrations determined and average annual consumption of food and water. The daily intakes (in mBq) were 22.1 (238U), 26.5 (234U), 2.38 (232Th), 4.06 (230Th), 11.2 (228Th) and 42.2 (226Ra). The intake of uranium isotopes occurred mainly with water; the main intake of thorium isotopes was with animal products, vegetables, cereals and potatoes, whereas 226Ra entered mainly with animal products, cereals and vegetables. From the intake and dose coefficients, the annual effective doses for the ingested radionuclides were calculated. The annual effective dose was 5.95 microSv, of which 72.4% originated from 226Ra. PMID- 11419600 TI - Indoor air quality at nine shopping malls in Hong Kong. AB - Hong Kong is one of the most attractive shopping paradises in the world. Many local people and international tourists favor to spend their time in shopping malls in Hong Kong. Good indoor air quality is, therefore, very essential to shoppers. In order to characterize the indoor air quality in shopping malls, nine shopping malls in Hong Kong were selected for this study. The indoor air pollutants included carbon dioxide (CO2), carbon monoxide (CO), total hydrocarbons (THC), formaldehyde (HCHO), respirable particulate matter (PM10) and total bacteria count (TBC). More than 40% of the shopping malls had 1-h average CO2 levels above the 1000 ppm of the ASHRAE standard on both weekdays and weekends. Also, they had average weekday PM10 concentrations that exceeded the Hong Kong Indoor Air Quality Objective (HKIAQO). The highest indoor PM10 level at a mall was 380 microg/m3. Of the malls surveyed, 30% had indoor airborne bacteria levels above 1000 cfu/m3 set by the HKIAQO. The elevated indoor CO2 and bacteria levels could result from high occupancy combined with insufficient ventilation. The increased PM10 levels could be probably attributed to illegal smoking inside these establishments. In comparison, the shopping malls that contained internal public transport drop-off areas, where vehicles were parked with idling engines and had major entry doors close to heavy traffic roads had higher CO and PM10 indoor levels. In addition, the extensive use of cooking stoves without adequate ventilation inside food courts could increase indoor CO2, CO and PM10 levels. PMID- 11419601 TI - Exergy analysis in the assessment of the sustainability of waste gas treatment systems. AB - This study focuses on the sustainability of different technological options for the treatment of waste gases from a waste water treatment plant loaded with volatile organic compounds. The options considered are biofiltration, active carbon adsorption and catalytic and thermal oxidation. The amount of resources and utilities to construct and operate each system have been investigated from the point of view of the Second Law of thermodynamics. The unit in which all resources are treated is Joules of exergy. It was concluded that biofiltration was the most exergetically efficient system. The cumulative exergy consumption of the resources and utilities for construction and operation have been quantified in exergy terms. Further on, the requirements for the abatement of emissions generated by operating the waste gas treatment systems and the amount of renewables have been taken into account in the assessment of the sustainability of the waste gas treatment technologies. Finally, a comparison between exergy analysis and life cycle analysis in assessing the sustainability of the waste gas treatment options, is presented. PMID- 11419602 TI - Organochlorine pesticide residues in human milk of a Hmong hill tribe living in Northern Thailand. AB - In December 1998 whole breast milk samples from 25 Hmong mothers living in the village of Mae Sa Mai, 40 km north of Chiang Mai City, Northern Thailand, were collected and analysed for DDT, heptachlor, HCB and HCH residues (fat normalized data). Short questionnaires and anthropometric measurements were used to obtain information on personal characteristics, lifestyle, contact with pesticides, dietary habits and former residences. DDT was detected in all samples with a median and maximum level of 209 and 2012 ng of total DDT isomers per millilitre of milk, respectively. The median and highest percentages of p,p'-DDT were 23.2 and 44.7%. In 15 samples heptachlor was detected in the metabolized form of heptachlor-epoxide with a median value of 4.4 ng/ml. The estimated daily intakes of DDT, heptachlor and heptachlor-epoxide by the infants exceeded up to 20 times the acceptable daily intakes as recommended by the FAO and WHO. In nine samples HCB was detected with a median value of 5.4 ng/ml from which seven of the nine also had the highest values of DDT residues. The gamma-isomer of HCH was only found in one sample with 3.6 ng/ml. The mean sum-DDT residues with 14.96 mg/kg milk fat, as well as the estimated daily intakes by the infants are one of the highest reported in the 1990s. The fact that the mother breast-feeds her first child and that she originally comes from a region where DDT is still in use as a vector control agent, as well as the former use of organochlorine pesticides (OCPs) in agriculture, seem to be the main factors for high DDT and other OCP residues in the mothers' milk. PMID- 11419603 TI - Heavy metals in the tissues of common dolphins (Delphinus delphis) stranded on the Portuguese coast. AB - The concentrations of Hg, Sn, Cr, Zn, Ni, Co, Cd, Mn, Fe and Cu were determined in the liver, kidney and muscle of 24 common dolphins stranded on the Portuguese coast between 1995 and 1998. Nitric acid was used to extract the metals from the tissues for analysis by inductively-coupled plasma/atomic emission spectroscopy (ICP/AES). Those metals with relatively higher concentrations included Fe, Zn and Hg, particularly in liver. Other metals including Cr, Ni and Cd tended to show much lower levels, or were even undetected (e.g. Co). Different metals seemed to show different temporal trends, although due to substantial variations of the mean concentrations for each year obtained, such temporal pattern has to be studied further. Total Hg concentration in the kidney, muscle and particularly liver, were higher in females than in males. Total Hg concentrations in all the organs increased with body length of dolphins, whilst those of Zn and Cu in muscle decreased with dolphin length. A strong correlation was found between essential metals Zn and Cu in muscle, possibly resulting from sequestration of these metals by metallothionein. In addition, significant co-associations existed between the same metal (e.g. Hg, Sn and Zn) in different organs. PMID- 11419604 TI - Lung cancer risks among lead smelter workers also exposed to arsenic. AB - Recent publications indicate an increased incidence of lung cancer in non-ferrous smelter workers exposed to lead. The present study provides further data on selected subgroups in one such cohort. The cohort was based on 3979 smelter workers employed for at least 1 year between 1928 and 1979, and also exposed to lead and included in the Blood Lead Register that was started at the smelter in 1950. Two subcohorts were formed from the original cohort. One consists of 710 workers employed at the lead departments (Lead subcohort 1), and the other of 383 workers employed at the lead departments (Lead subcohort 2), but never at other works where an excess lung cancer risk was previously identified. Standardized Cancer Incidence Rates (SIR) 1958-1987 were calculated relative to county rates. In the subcohorts, arsenic exposure in lung cancer cases was assessed in detail based on occupational hygiene information from the company. Lung cancer incidence was raised in both subcohorts (Lead subcohort 1: SIR 2.4; 95% CI 1.2-4.5; Lead subcohort 2: SIR 3.6; 95% CI 1.2-8.3). Total cancer incidence in the cohorts was not increased. A detailed study of arsenic exposure in the 10 lung cancer cases in these two subcohorts revealed that all but one of these cases had a significant exposure also to arsenic. An elevated incidence of lung cancer was observed in smelter workers exposed to lead. However, considerable arsenic exposure also occurred in a majority of the lung cancer cases in the investigated subcohorts. In this multifactorial exposure situation it has not been possible to separate the carcinogenic effects of lead and arsenic, but a possible interaction between these metals may be involved in explaining the carcinogenic risks. PMID- 11419605 TI - Heat treatment of incinerator retired sorbents containing heavy metals. AB - Previous studies have indicated that an effective control technique for heavy metal emissions from incinerators is to use solid sorbents to capture metals by physical deposition and chemical adsorption. However, the adsorbed heavy metals on the retired sorbents could possibly cause secondary pollution if they are inadequately treated. The main objective of this study is to increase the stability of four heavy metals (Cu, Pb, Cr and Cd) on retired sorbents by heat treatment. The stability was based on the toxicity characteristic leaching procedure (TCLP) tests. The parameters evaluated included: (1) different temperatures (900, 1100 and 1300 degrees C); and (2) various heating times (20, 40 and 60 min). The results indicated that most of the leaching rates decreased with increasing temperature for the four toxic heavy metals. The heat treatment time did not regularly influence the stability of heavy metals, and affected it slightly when the temperature was controlled at 1300 degrees C. PMID- 11419606 TI - Inhibition of human plasmin activity using humic acids with arsenic. AB - In this study, we examined the effects of natural humic acid (HA) purified from drinking well-water in Blackfoot disease (BFD) endemic areas, using synthetic humic acid (SHA), such as protocatechuic acid, ferulic acid, vanillic acid or catechol, and trivalent arsenic on human plasmin activity. Data in this report indicated that both HA and SHA inhibited human plasmin activity by 20-80 and 5 95%, respectively, at concentrations of 20-480 microg/ml. Organometallic complexes composed of HA and arsenic show enhanced inhibition of plasmin activity as compared with either HA or arsenic alone. Monomers of HA or arsenic alone do not inhibit plasma activity. Oxidative stress may play a role in the inhibition of plasma activity, as various free-radical scavengers, such as ascorbic acid, alpha-tocopherol, catalase and superoxide dismutase (SOD), abrogate the inhibitory effects of HA and HA-arsenic complexes. The notion that HA/organometallic complexes (HA/OR) impaired plasmin activity was significant, due to the fact that both of these agents (HA and arsenic) are etiological factors in the development of peripheral vascular diseases, such as BFD. This report substantiates the inhibitory effects of HA/OR on plasmin activity, and thus provides evidence for the partial mechanism of action of HA/OR in BFD. PMID- 11419607 TI - Thought-action fusion and thought suppression in obsessive-compulsive disorder. AB - To examine the significance of thought-action fusion (TAF) and thought suppression tendencies, the present study obtained pre- and post-treatment questionnaire data on these constructs in a sample of OCD patients (n=24) and non OCD anxiety patients (n=20). Results indicate that TAF and suppression are correlated with severity of psychopathology. Yet, the associations between TAF and psychopathology are not typical for OCD, but do also occur in other anxiety disorders (e.g., panic disorder, post traumatic stress disorder, and social phobia). As well, mean scores on the TAF and thought suppression measures dropped significantly from pre- to post-treatment, indicating that TAF and thought suppression are susceptible to change during psychotherapy. PMID- 11419608 TI - Thought-action fusion in individuals with OCD symptoms. AB - Rachman (Rachman, S. (1993). Obsessions, responsibility, and guilt. Behaviour Research and Therapy, 31, 149-154) suggested that patients with OCD may interpret thoughts as having special importance, thus experiencing thought-action fusion (TAF). Shafran, Thordarson and Rachman (Shafran, R., Thordarson, D. S. & Rachman, S. (1996). Thought-action fusion in obsessive compulsive disorder. Journal of Anxiety Disorders, 710, 379-391) developed a questionnaire (TAF) and found that obsessives scored higher than non-obsessives on the measure. In the current study, we modified the TAF to include a scale that assessed the "likelihood of events happening to others" as well as ratings of the responsibility and cost for having these thoughts. Replicating previous findings, we found that individuals with OC symptoms gave higher ratings to the likelihood of negative events happening as a result of their negative thoughts. Individuals with OC symptoms also rated the likelihood that they would prevent harm by their positive thoughts higher than did individuals without OC symptoms. These results suggest that the role of thought-action fusion in OCs may extend to exaggerated beliefs about thoughts regarding the reduction of harm. PMID- 11419609 TI - Determinants of early- vs late-onset dental fear in a longitudinal epidemiological study. AB - A longitudinal investigation of risk factors for early- and late-onset dental fear was conducted. Early-onset dental fear was related to conditioning experiences (indexed via caries level and tooth loss), service use patterns, stress reactive personality and specific beliefs about health professionals. Late onset dental fear was related to aversive conditioning experiences, irregular service use and an external locus of control. In contrast to recent findings for dental anxiety, personality factors were not strongly related to the onset of dental fear in young adulthood. The key role played by conditioning events in the development of both early- and late-onset dental fear was confirmed. Conditioning events appear to play a different role in the development of dental fear vs dental anxiety. This may reflect important, but largely ignored differences between these two closely-related constructs. Interventions for early-onset dental fear should aim to modify both the dental fear and the personality vulnerabilities that may contribute to the development of dental fear early in the life-course. PMID- 11419610 TI - Chronic pain: a reformulation of the cognitive-behavioural model. AB - The last few decades has seen psychological theories of chronic pain dominated by behavioural concepts and particularly, by the operant model as described by Fordyce et al. (1968: Fordyce, W., Fowler, R., & DeLateur, B. (1968) An application of behavior modification technique to a problem of chronic pain. Behaviour Research and Therapy, 6, 105-107, 1976: Fordyce, W.E. (1976) Behavioral methods for chronic pain and illness. St Louis, MO: C. V Moseby). More recently, cognitive constructs have been included, giving the impression that the operant theory of chronic pain has been replaced by a cognitive-behavioural theory (CBT). Given the fact that CBTs were introduced essentially to overcome some inherent problems with operant theory, it is notable that they are still very much based on operant theory and continue to be founded upon operant principles. Further, the extent to which CBTs (at this stage) have included contemporary cognitive research has been limited. This paper argues that it is of questionable validity to continue to include problematic concepts (e.g., operant principles) into a revised theory (e.g., CBT). Instead, consistent with research from other areas (particularly the anxiety disorders), chronic pain and the problems associated with it may be better explained by a reformulated cognitive-behavioural theory which although not ignoring the observable behaviours of pain patients and their associations with social reinforcers, interprets these phenomena from a cognitive perspective. Thus, a modified CBT is proffered, focusing more directly on patients' thoughts about, and appraisals of, their pain. Evidence in support of such a theory is provided, as are suggestions for further research and the implications such a theory has for treatment. PMID- 11419611 TI - The effects of relaxation response meditation on the symptoms of irritable bowel syndrome: results of a controlled treatment study. AB - In this study, Herbert Benson's (1975) Relaxation Response Meditation program was tested as a possible treatment for Irritable Bowel Syndrome (IBS). Participants were 16 adults who were matched into pairs based on presence of Axis I disorder, primary IBS symptoms and demographic features and randomized to either a six week meditation condition or a six week wait list symptom monitoring condition. Thirteen participants completed treatment and follow-up. All subjects assigned to the Wait List were subsequently treated. Patients in the treatment condition were taught the meditation technique and asked to practice it twice a day for 15 minutes. Composite Primary IBS Symptom Reduction (CPSR) scores were calculated for each patient from end of baseline to two weeks post-treatment (or to post wait list). One tailed independent sample t-tests revealed that Meditation was superior to the control (P=0.04). Significant within-subject improvements were noted for flatulence (P=0.03) and belching (P=0.02) by post-treatment. By three month follow-up, significant improvements in flatulence (P<0.01), belching (P=0.02), bloating (P=0.05), and diarrhea (P=0.03) were shown by symptom diary. Constipation approached significance (P=0.07). Benson's Relaxation Response Meditation appears to be a viable treatment for IBS. PMID- 11419612 TI - Memory bias, confidence and responsibility in compulsive checking. AB - Recent research suggests that there is a positive memory bias for threatening information in compulsive cleaners. However, the relationship between OCD and memory is likely to be more complex when the compulsive behaviour is checking. Hence, we decided to explore this relationship in a clinical sample of people who check compulsively. Participants completed a diagnostic interview and were then asked to complete a standard 'baseline' check which normally causes distress/discomfort. Two additional checks were then completed--one under conditions of high responsibility and one under low responsibility. The order of responsibility manipulation was randomized across participants. After each check, participants completed a Memory and Confidence Interview which assessed memory for threat-relevant and threat-irrelevant aspects of the check, and also confidence in memory for the check. One week later, participants came into the laboratory to complete additional Memory and Confidence Interviews after watching a videotape of the checks completed earlier in their own homes. These videotaped checks were taken as conditions of 'no responsibility'. Results show a positive memory bias for threat-relevant information. As responsibility was inflated, this positive memory bias was amplified. Under conditions of no responsibility, no memory bias was detectable. Also, responsibility appears to have had a greater impact on confidence in memory than on memory itself in OCD. The results are discussed in terms of the mnestic deficit theory of OCD and in terms of cognitive behavioural approaches to understanding the disorder. PMID- 11419613 TI - Evaluation of a short-term group therapy program for children with behavior problems and their parents. AB - The current study investigated the effectiveness of a short-term, cognitive behavioral program for 106 primary school-aged children referred with externalizing behavior problems and their parents, compared with 39 children and their parents on a waiting-list to be treated. Exploring Together' comprised a children's group (anger management, problem-solving and social skills training), a parents' group (parenting skills training and dealing with parents' own issues), and a combined children's and parents' group (to target parent-child interactions). The program reduced children's behavior problems and improved their social skills at home. Changes in children's behaviors and social skills at home were generally maintained at 6- and 12-month follow-up. Implications of the findings for improving interventions for childhood externalizing behavior problems are discussed. PMID- 11419614 TI - Thought-action fusion and anxiety disorders symptoms in normal adolescents. AB - The present study examined thought-action fusion (TAF) in a large sample of normal adolescents (n=427). Participants completed the Thought-Action Fusion Questionnaire for Adolescents (TAFQ-A) and scales measuring trait anxiety, symptoms of obsessive-compulsive disorder, other anxiety disorders, and depression. Results showed that the TAFQ-A is a reliable instrument assessing two dimensions of TAF, viz. Morality (i.e., the belief that unacceptable thoughts are morally equivalent to overt actions) and Likelihood (i.e., the belief that thinking of an unacceptable or disturbing situation will increase the probability that that situation actually occurs). Furthermore, TAF was not only associated with symptoms of OCD, but also with symptoms of other anxiety disorders and depression. However, when controlling for levels of trait anxiety, most connections between TAF and anxiety disorders symptoms disappeared. Symptoms of OCD and generalised anxiety remained significantly related to TAF. Altogether, the data are supportive of the notion that TAF is involved in a broad range of anxiety disorders and in particular OCD. PMID- 11419615 TI - Psychometric evaluation on the Japanese adaptation of the Aggression Questionnaire. AB - The psychometric properties of a Japanese translation of the Aggression Questionnaire (Buss & Perry, 1992, Journal of Personality and Social Psychology, 63, 452-459) were investigated. Factor analysis of the responses of 425 Japanese participants generally supported the four-factor model. The factors were Physical Aggression, Verbal Aggression, Anger and Hostility. The internal consistencies of the four subscales were adequate. The results suggested that the Japanese version of the Aggression Questionnaire met psychometric standards and appears to be a promising measure of aggression. However, the Japanese version may be improved if two reversed scored items are removed from the scale. The cross-cultural difference in responses between negatively-oriented and affirmatively-oriented questions was discussed. PMID- 11419616 TI - Dimensionality and correlates of problem solving: the use of the Problem Solving Inventory in the Chinese context. AB - The dimensional structure of the Problem Solving Inventory (PSI) was investigated using data collected from a sample of 499 Chinese university students in teacher training. While the original three-factor model provided poor fit to the data, the revised two-factor model provided more adequate fit but failed to consider a PSI construct tapped in the original PSI formulation. The newly constructed PSI scales based on the present data provided viable alternative scales reflecting constructs of self-efficacy, rational coping and dysfunctional coping. These newly developed problem-solving scales were predictive of indices of psychological adjustment in terms of depression, burnout, and perceived self efficacy toward helping others. The use of the PSI in the Chinese context is discussed. PMID- 11419617 TI - Spatio-temporal pattern formation on spherical surfaces: numerical simulation and application to solid tumour growth. AB - In this paper we examine spatio-temporal pattern formation in reaction-diffusion systems on the surface of the unit sphere in 3D. We first generalise the usual linear stability analysis for a two-chemical system to this geometrical context. Noting the limitations of this approach (in terms of rigorous prediction of spatially heterogeneous steady-states) leads us to develop, as an alternative, a novel numerical method which can be applied to systems of any dimension with any reaction kinetics. This numerical method is based on the method of lines with spherical harmonics and uses fast Fourier transforms to expedite the computation of the reaction kinetics. Numerical experiments show that this method efficiently computes the evolution of spatial patterns and yields numerical results which coincide with those predicted by linear stability analysis when the latter is known. Using these tools, we then investigate the rjle that pre-pattern (Turing) theory may play in the growth and development of solid tumours. The theoretical steady-state distributions of two chemicals (one a growth activating factor, the other a growth inhibitory factor) are compared with the experimentally and clinically observed spatial heterogeneity of cancer cells in small, solid spherical tumours such as multicell spheroids and carcinomas. Moreover, we suggest a number of chemicals which are known to be produced by tumour cells (autocrine growth factors), and are also known to interact with one another, as possible growth promoting and growth inhibiting factors respectively. In order to connect more concretely the numerical method to this application, we compute spatially heterogeneous patterns on the surface of a growing spherical tumour, modelled as a moving-boundary problem. The numerical results strongly support the theoretical expectations in this case. Finally in an appendix we give a brief analysis of the numerical method. PMID- 11419618 TI - A deterministic size-structured population model for the worm Naidis elinguis. AB - In this paper we model the population dynamics of the worm Nais elinguis, which reproduces by division into two unequal parts. By using renewal theory we derive the asymptotic behaviour of a Naidis elinguis population. In particular we prove a certain relation between the fraction of the population that was born small (respectively the fraction that was born large) and the inter-division times. PMID- 11419619 TI - The effect of dispersal on single-species nonautonomous dispersal models with delays. AB - In this paper, single-species nonautonomous dispersal models with delays are considered. An interesting result on the effect of dispersal for persistence and extinction is obtained. That is, if the species is persistent in a patch then it is also persistent in all other patches; if the species is permanent in a patch then it is also permanent in all other patches; if the species is extinct in a patch then it is also extinct in all other patches. Furthermore, some new sufficient conditions for the permanence and extinction of the species in a patch are established. The existence of positive periodic solutions is obtained in the periodic case by employing Teng and Chen's results on the existence of positive periodic solutions for functional differential equations. PMID- 11419620 TI - Mutation and recombination with tight linkage. AB - An exact solution of the mutation-recombination equation in continuous time is presented, with linear ordering of the sites and at most one mutation or crossover event taking place at every instant of time. The differential equation may be obtained from a mutation-recombination model with discrete generations, in the limit of short generations, or weak mutation and recombination. The solution relies on the multilinear structure of the dynamical system, and on the commuting properties of the mutation and recombination operators. It is obtained through diagonalization of the mutation term, followed by a transformation to certain measures of linkage disequilibrium that simultaneously linearize and diagonalize the recombination dynamics. The collection of linkage disequilibria, as well as their decay rates, are given in closed form. PMID- 11419621 TI - Angular weighted hybrid cone-beam CT reconstruction for circular trajectories. AB - Hybrid reconstruction techniques have been introduced for the volume reconstruction of axially truncated cone-beam computed tomography projection data acquired along a circular source-detector trajectory. The introduction of weighted half-scan techniques into this framework is described in this paper. Due to the cone-beam geometry it is not possible to perform the weighting on the projections as is typically done in conventional single-line computed tomography. Hence, in this paper we present an efficient way to incorporate angular weighting functions, depending on the object point position, into the framework of hybrid cone-beam reconstruction. Four different angular weighting functions are introduced and discussed with respect to their cone-beam artefact behaviour and their influence on the signal-to-noise ratio. As a result, the most effective angular weighting function for hybrid circular cone-beam reconstruction is determined by means of a simulation study based on mathematical phantoms and clinical data sets. This distance-weighted angular weighting scheme yields the best results in terms of high image quality, low computational complexity and signal-to-noise variations in the reconstruction volume. PMID- 11419622 TI - Conductivity of living intracranial tissues. AB - Resistivity values were measured from living human brain tissue in nine patients. A monopolar needle electrode was used with a measurement frequency of 50 kHz. Mean values were 3.51 Ohms m for grey matter and 3.91 Ohms m for white matter. Cerebrospiral fluid had a mean value of 0.80 Ohms m. Values for tumour tissues were dependent on the type of tumour and ranged from 2.30 to 9.70 Ohms m. PMID- 11419623 TI - Changes in the dielectric properties of rat tissue as a function of age at microwave frequencies. AB - The dielectric properties of ten rat tissues at six different ages were measured at 37 degrees C in the frequency range of 130 MHz to 10 GHz using an open-ended coaxial probe and a computer controlled network analyser. The results show a general decrease of the dielectric properties with age. The trend is more apparent for brain, skull and skin tissues and less noticeable for abdominal tissues. The variation in the dielectric properties with age is due to the changes in the water content and the organic composition of tissues. The percentage decrease in the dielectric properties of certain tissues in the 30 to 70 day old rats at cellular phone frequencies have been tabulated. These data provide an important input in the provision of rigorous dosimetry in lifetime exposure animal experiments. The results provide some insight into possible differences in the assessment of exposure for children and adults. PMID- 11419624 TI - The practical application of signal detection theory to image quality assessment in x-ray image intensifier-TV fluoroscopy. AB - This paper applies a published version of signal detection theory to x-ray image intensifier fluoroscopy data and compares the results with more conventional subjective image quality measures. An eight-bit digital framestore was used to acquire temporally contiguous frames of fluoroscopy data from which the modulation transfer function (MTF(u)) and noise power spectrum were established. These parameters were then combined to give detective quantum efficiency (DQE(u)) and used in conjunction with signal detection theory to calculate contrast-detail performance. DQE(u) was found to lie between 0.1 and 0.5 for a range of fluoroscopy systems. Two separate image quality experiments were then performed in order to assess the correspondence between the objective and subjective methods. First, image quality for a given fluoroscopy system was studied as a function of doserate using objective parameters and a standard subjective contrast-detail method. Following this, the two approaches were used to assess three different fluoroscopy units. Agreement between objective and subjective methods was good; doserate changes were modelled correctly while both methods ranked the three systems consistently. PMID- 11419625 TI - An SVM classifier to separate false signals from microcalcifications in digital mammograms. AB - In this paper we investigate the feasibility of using an SVM (support vector machine) classifier in our automatic system for the detection of clustered microcalcifications in digital mammograms. SVM is a technique for pattern recognition which relies on the statistical learning theory. It minimizes a function of two terms: the number of misclassified vectors of the training set and a term regarding the generalization classifier capability. We compare the SVM classifier with an MLP (multi-layer perceptron) in the false-positive reduction phase of our detection scheme: a detected signal is considered either microcalcification or false signal, according to the value of a set of its features. The SVM classifier gets slightly better results than the MLP one (Az value of 0.963 against 0.958) in the presence of a high number of training data; the improvement becomes much more evident (Az value of 0.952 against 0.918) in training sets of reduced size. Finally, the setting of the SVM classifier is much easier than the MLP one. PMID- 11419626 TI - Photothermal coagulation of blood vessels: a comparison of high-speed optical coherence tomography and numerical modelling. AB - Optical-thermal models that can accurately predict temperature rise and damage in blood vessels and surrounding tissue may be used to improve the treatment of vascular disorders. Verification of these models has been hampered by the lack of time- and depth-resolved experimental data. In this preliminary study, an optical coherence tomography system operating at 4-30 frames per second was used to visualize laser irradiation of cutaneous (hamster dorsal skin flap) blood vessels. An argon laser was utilized with the following parameters: pulse duration 0.1-2.0 s, spot size 0.1-1.0 mm, power 100-400 mW. Video microscopy images were obtained before and after irradiations, and optical-thermal modelling was performed on two irradiation cases. Time-resolved optical coherence tomography and still images were compared with predictions of temperature rise and damage using Monte Carlo and finite difference techniques. In general, predicted damage agreed with the actual blood vessel and surrounding tissue coagulation seen in images. However, limitations of current optical-thermal models were identified, such as the inability to model the dynamic changes in blood vessel diameter that were seen in the optical coherence tomography images. PMID- 11419627 TI - Mathematical model of antibody targeting: important parameters defined using clinical data. AB - Antibody-targeted therapy of cancer has shown benefits in the treatment of some cancers but selective delivery has not been optimized. Many parameters influence antibody targeting; some will have a greater effect than others and their effects will generally be interrelated. They include effects of blood flow and pressure, vascular permeability, venous and lymphatic drainage, permeation through extravascular spaces, antibody clearance, specificity, affinity and resistance to degradation. Quantitative data about the behaviour of targeting systems can be collected, and it is possible to describe the system in terms of compartments interconnected by equations defining the passage of targeting agents between them. A mathematical model of antibody targeting can thus be built. We have collected data on the time course of the distribution of four different antibody molecules of molecular weight 27, 100 and 150 kDa directed against carcinoembryonic antigen in patients with colorectal cancer. Laboratory data were used for parameters which could not be measured in patients. These data have been used to test the validity of the model for man and to develop it so that it is consistent with the diverse clinical data. The model is then used to understand the effects of changes to a parameter on tumour targeting efficiency and to select those parameters which have the greatest effect in therapy. Affinity of antibody, flow of antibody through the tumour and rate of elimination of antibody from the tumour were shown to be the most powerful parameters determining antibody localization. These concepts can be used to determine design parameters for antibody-targeted cancer therapy. PMID- 11419628 TI - Intensity modulated irradiation of a thorax phantom: comparisons between measurements, Monte Carlo calculations and pencil beam calculations. AB - The present study investigates the application of compensators for the intensity modulated irradiation of a thorax phantom. Measurements are compared with Monte Carlo and standard pencil beam algorithm dose calculations. Compensators were manufactured to produce the intensity profiles that were generated from the scientific version of the KonRad IMRT treatment-planning system for a given treatment plan. The comparison of dose distributions calculated with a pencil beam algorithm, with the Monte Carlo code EGS4 and with measurements is presented. By measurements in a water phantom it is demonstrated that the method used to manufacture the compensators reproduces the intensity profiles in a suitable manner. Monte Carlo simulations in a water phantom show that the accelerator head model used for simulations is sufficient. No significant overestimations of dose values inside the target volume by the pencil beam algorithm are found in the thorax phantom. An overestimation of dose values in lung by the pencil beam algorithm is also not found. Expected dose calculation errors of the pencil beam algorithm are suppressed, because the dose to the low density region lung is reduced by the use of a non-coplanar beam arrangement and by intensity modulation. PMID- 11419629 TI - Calculation of perturbation correction factors for some reference dosimeters in high-energy photon beams with the Monte Carlo code PENELOPE. AB - The BNM-LNHB (formerly BNM-LPRI, the French national standard laboratory for ionizing radiation) is equipped with a SATURNE 43 linear accelerator (GE Medical Systems) dedicated to establishing national references of absorbed dose to water for high-energy photon and electron beams. These standards are derived from a dose measurement with a graphite calorimeter and a transfer procedure to water using Fricke dosimeters. This method has already been used to obtain the reference of absorbed dose to water for cobalt-60 beams. The correction factors rising from the perturbations generated by the dosimeters were determined by Monte Carlo calculations. To meet these applications, the Monte Carlo code PENELOPE was used and user codes were specially developed. The first step consisted of simulating the electron and photon showers produced by primary electrons within the accelerator head to determine the characteristics of the resulting photon beams and absorbed dose distributions in a water phantom. These preliminary computations were described in a previous paper. The second step, described in this paper, deals with the calculation of the perturbation correction factors of the graphite calorimeter and of Fricke dosimeters. To point out possible systematic biases, these correction factors were calculated with another Monte Carlo code, EGS4, widely used for years in the field of dose metrology applications. Comparison of the results showed no significant bias. When they were possible, experimental verifications confirmed the calculated values. PMID- 11419630 TI - Analytical calculation of the portal scatter to primary dose ratio: an EGS4 Monte Carlo and experimental validation at large air gaps. AB - An analytical approximation for the scatter to primary dose ratio (SPR) on the central axis was validated against Monte Carlo results and experimental measurements for homogeneous and inhomogeneous phantoms. The analytical approximation only included first-order Compton scatter. The contribution to the total SPR from first-order Compton scatter, multiply scattered photons and electron scatter was investigated using Monte Carlo simulation for homogeneous phantoms (up to 30 cm thick for 6 and 18 MV beams; source to detector distances from 150 to 230 cm) as well as for a neck, thorax and pelvis phantom. SPRs were measured on the central axis with an ionization chamber for water phantoms (up to 20 cm thick at 4 MV, 30 cm for 6 MV and 10 MV and 40 cm for 18 MV; source to detector distances of 185 and 200 cm) and for phantoms representing the neck, thorax and pelvis (for air gaps of 50 cm and larger). The mean difference between the experimental and analytical SPRs on the central axis for source to detector distances of 170 cm or greater was within: -0.003 (neck); -0.012 (thorax); -0.028 (pelvis, 10 MV) and 0.008 (pelvis, 18 MV) respectively. PMID- 11419631 TI - Comment on genetic and global algorithms for optimization of three-dimensional conformal radiotherapy treatment planning. PMID- 11419632 TI - Cone-beam image reconstruction using spherical harmonics. AB - Image reconstruction from cone-beam projections is required for both x-ray computed tomography (CT) and single photon emission computed tomography (SPECT). Grangeat's algorithm accurately performs cone-beam reconstruction provided that Tuy's data sufficiency condition is satisfied and projections are complete. The algorithm consists of three stages: (a) Forming weighted plane integrals by calculating the line integrals on the cone-beam detector, and obtaining the first derivative of the plane integrals (3D Radon transform) by taking the derivative of the weighted plane integrals. (b) Rebinning the data and calculating the second derivative with respect to the normal to the plane. (c) Reconstructing the image using the 3D Radon backprojection. A new method for implementing the first stage of Grangeat's algorithm was developed using spherical harmonics. The method assumes that the detector is large enough to image the whole object without truncation. Computer simulations show that if the trajectory of the cone vertex satisfies Tuy's data sufficiency condition, the proposed algorithm provides an exact reconstruction. PMID- 11419633 TI - A rapid procedure for initial drug evaluation. AB - The overall aim of this work is to develop computer simulations to aid in the selection of proposed medicines and identify those most likely to succeed. One important feature is a systems approach to simulate both the target area with which the drug is designed to interact as well as the surrounding areas where feedback mechanisms may alter the expected effect. The simulation must be rapid if it is to be used to evaluate large numbers of potential drugs. Thus the procedure simplifies many of the known complex phenomena to provide a general framework and feedback mechanisms. An example of the use of the simulation to study a drug used to treat hypertension is given. A possible use of the technique is shown using the example of the effect of varying the drug dosage on the contraction of the arteriole muscle. PMID- 11419634 TI - Temperature measurement artefacts of thermocouples and fluoroptic probes during laser irradiation at 810 nm. AB - This study examined the artefact induced in temperature measurements made with thermocouples and Luxtron fluoroptic probes in the presence of infrared radiation. Localized heating was created using a continuous-wave, 810 nm diode laser system emitting 2.0 W from a cylindrical diffusing optical fibre, in air, water and an agar-albumin phantom. The temperature was measured every 1.0 s for 10 to 150 s, with both a thermocouple and a Luxtron fluoroptic probe at distances of 2, 3, 4, 5, 6 and 7 mm from the cylindrical diffusing tip. In all cases, the fluoroptic probe recorded a higher temperature than the thermocouple during laser irradiation. The difference in measured temperatures between the Luxtron probe and the thermocouple ranged from 1.6 degrees C to 18.8 degrees C in air, from 0.3 degrees C to 10.2 degrees C in water, and from 1.4 degrees C to 10.1 degrees C in phantom, depending on the distance of the probe from the laser source. The results suggest that in the presence of laser irradiation, self-heating of the Luxtron probe induces a significant artefact in temperature measurements at distances less than 4 mm from the source fibre. As a result, fluoroptic probes may not be suitable for monitoring tissue temperature for treatments when laser irradiation is present if sensors are located close to the fibre tip (<4 mm). PMID- 11419635 TI - Recording accelerator monitor units during electronic portal imaging: application to collimator position verification during IMRT. AB - The application of multiple portal image acquisition to collimator position verification during dynamic multileaf collimation (DMLC) using a commercial camera-based electronic portal imaging device (EPID) (Theraview, Cablon Medical BV, Leusden, The Netherlands) mounted on an Elekta SL15i accelerator (Elekta Oncology Systems, Crawley, UK) is described. This is achieved using a custom built dose acquisition system optically interfaced to both the camera control unit of the EPID and the monitor unit (MU) channel of the accelerator. The method uses the beam blanking camera control signal to trigger the dose acquisition system to read the cumulative accelerator MUs at the beginning and end of each period of image formation. A maximum delay of 15 ms has been estimated for recording of accelerator MUs in the current system. The camera interface was observed to have no effect on the operation of the EPID during normal clinical use and could therefore be left permanently in situ. Use of the system for collimator position verification of a test case is presented. The technique described uses a specific camera-based EPID and accelerator, although the general principle of using an EPID control signal to trigger recording of accelerator MUs may be applicable to other EPIDs/accelerators with suitable knowledge of the accelerator dosimetry system. PMID- 11419636 TI - Feasibility study of an online toxicological sensor based on the optical waveguide technique. AB - Morphological properties of the cells often change as an early response to the presence of a pharmacologically acting toxic substance [Etcheverry, S.B., Crans, D.C., Keramidas, A.D., Cortizo, A.M., Arch. Biochem. Biophys. 338 (1997) 7-14]. Recently it has been shown that living animal cell adhesion and spreading can be monitored online and quantitatively via the interaction of the cells with the evanescent electromagnetic field present at the surface of an optical waveguide [Ramsden, J.J., Li, S.Y., Heinzle, E., Prinosil, J.E. Cytometry 19 (1995) 97 102]. In the present study, optical waveguide lightmode spectroscopy (OWLS) and confocal laser scanning microscopy (CLSM), which provides information about the shape of the cells at the surface, were compared under identical experimental conditions. This allowed for the correlation between the cell-shape information from CLSM and the cell-surface interaction measurements from OWLS. The proposed design of the microsystem sensor involves the establishment of a cell layer on the surface of the waveguide and the subsequent online measurement of the morphological response of the cells to various toxic substances. In the present study, the setup was evaluated using cells from an osteoblastic MC 3T3-E1 cell line, and sodium hypochlorite was used as model toxic substance. Comparing the OWLS signal to the morphological response measured by CLSM reveals that OWLS is effective in monitoring not only cell attachment and spreading but also the cellular response to toxic compounds (i.e. by means of change in cell morphology). For the model toxin, the OWLS measurements indicate that, at concentrations above 0.01%, the cells exhibit a clearly discernable morphological effect (i.e. a decrease in average cell contact area). Thus, the potential of an on-line sensor based on OWLS to applications in toxicology, pharmacy and biocompatibility was demonstrated. PMID- 11419637 TI - Ferrocene-avidin conjugates for bioelectrochemical applications. AB - Coupling of ferrocene moieties to avidin via a flexible spacer molecule yields a conjugate which combines the unique biotin-binding properties of avidin with the reversible redox characteristics of ferrocenes. Synthesis of the conjugate has been optimised and the conjugates were characterised bio- and electrochemically. Covalent immobilisation of the conjugate on gold electrodes in a dense monolayer results in electrodes with a high binding capacity for biotinylated molecules as well as good electron transfer properties. The application potential of such electrodes for bioelectrochemical systems is demonstrated by electrochemical reduction of hydrogen peroxide under mild conditions catalysed by a bound biotin microperoxidase MP11 conjugate. PMID- 11419638 TI - A minisensor for the rapid screening of sucralose based on surface-stabilized bilayer lipid membranes. AB - This work describes an electrochemical technique that is suitable for the rapid and sensitive screening of the sweetener sucralose based on surface-stabilized bilayer lipid membranes (s-BLMs) composed of egg phosphatidylcholine. The interactions of sucralose with s-BLMs produced electrochemical ion current increases, which appeared reproducible within a few seconds after exposure of the membranes to the sweetener. The mechanism of signal generation was investigated by differential scanning calorimetric studies. The mechanism was found to be associated with alteration of the electrostatic fields of the lipid film. These studies revealed that an increase of the molecular area of the lipids at the membranes and a stabilization of a gel phase structure occurred due to adsorption of the sweetener. Water molecules are adsorbed at the polar headgroups of the lipids, which changes the electrostatic field at the surface of the membranes. The current signal increases were related to the concentration of sucralose in bulk solution in the micromolar range. The present lipid film based sensor provided a fast response (i.e. in the order of a few seconds) to alterations of sucralose concentration (5-50 microm) in electrolyte solution. The electrochemical transduction of the interactions of this artificial sweetener with s-BLMs was applied in the determination of this compound in granulated sugar substitute products using the present minisensor. PMID- 11419639 TI - Amperometric glucose biosensors based on Prussian Blue- and polyaniline-glucose oxidase modified electrodes. AB - The properties of glucose sensors fabricated by immobilization of glucose oxidase in a layer of electrochemically deposited polyaniline were investigated. Selective amperometric glucose sensors were prepared by immobilization of glucose oxidase on a Prussian Blue-modified platinum electrode in a layer of polyaniline during a one-step electropolymerization procedure from phosphate buffer. The influence of ascorbic acid and acetaminophen was completely eliminated due to impermeability of polyaniline to these substances. PMID- 11419640 TI - Nanoporous anti-fouling silicon membranes for biosensor applications. AB - The ability to create biocompatible well-controlled membranes has been an area of great interest over the last few years, particularly for biosensor applications. The present study describes the fabrication and characterization of novel nanoporous micromachined membranes that exhibit selective permeability and low biofouling. Results indicate that such membranes can be fabricated with uniform pore sizes capable of the simultaneous exclusion of albumin and diffusion of glucose. Compared to polymeric membranes of similar pore size, micromachined silicon membranes allowed more than twice the amount of glucose diffusion after 240 min and complete albumin exclusion. Moreover, membranes exhibit no morphological change or degradability in the presence of biological proteins and fluids at 37 degrees C. The results point to the potential of using such membranes for implantable biosensor applications. With monodisperse pores sizes as small as 10 nm, these membranes offer advantages in their reproducibility, stability, and ability to be integrated in silicon-based biosensing technology. PMID- 11419641 TI - Structure and activity of lipid membrane biosensor surfaces studied with atomic force microscopy and a resonant mirror. AB - Three variants of the liposome fusion (coalescence) method to produce supported lipid bilayers, containing the ganglioside GM1 on silicon nitride surfaces, were studied. The first procedure involved attachment and fusion of liposomes containing DMPC, GM1 and a small amount of biotinylated lipid (Biotin-LC-DPPE) to a streptavidin coated surface. Direct fusion of liposomes composed of a mixture of DPPC, DPPG, DPPE, GM1 and cholesterol to the surface were the second variant. The final method utilised the second type of liposomes, fused onto a streptavidin layer with a small amount of exposed hydrophobic tails. The methods produced similar lipid layers, but with different ways of attachment to the surface. The binding of cholera toxin B-subunit (CTB) towards these sensor surfaces was measured in a resonant mirror biosensor instrument and the activity and longer term stability of the layers were examined. The prepared surfaces were also imaged by atomic force microscopy (AFM) in liquid to characterise the topography of the lipid layers. The binding efficiency of CTB towards these surfaces was discussed in terms of lipid fluidity and surface roughness. PMID- 11419642 TI - A high-density poly(ethylene glycol) polymer brush for immobilization on glass type surfaces. AB - Label-free heterogeneous phase detection critically depends on the properties of the interfacial layer. We have obtained high-density monomolecular poly(ethylene glycol) (PEG) layers by solvent-free coupling of homo-bifunctional PEGs (2,000 g/mol) at 75 degrees C to silica surfaces silanized with glycidyloxipropyltrimethoxysilane (GOPTS). Characterization by ellipsometry and contact angles revealed that PEG layers up to 3.4 ng/mm2 with low roughness and flexibility were obtained. Specific and non-specific binding at these PEG surfaces was monitored by reflectometric interference spectroscopy (RIfS). No significant non-specific adsorption upon incubation of 1 mg/ml ovalbumin was detectable (< 10 pg/mm2), and 150 pg/mm2 upon incubation of 10% calf serum, less than 10% of the amount adsorbed to the solely silanized surfaces. The terminal functional groups of the PEG layers were utilized to couple ligands and a protein. Specific protein interaction with these immobilized compounds was detected with saturation loadings in the range of protein monolayers (2-4 ng/mm2). The excellent functional properties, the high stability of the layers, the generic and practical coupling procedure and the versatility for immobilizing compounds of very different functionality make these PEG layers very attractive for application in label-free detection with silica or metal-oxide based transducers. PMID- 11419643 TI - Tryptophan phosphorescence as a monitor of protein conformation in molecular films. AB - This report enquires on the potentiality of Trp phosphorescence for probing the conformational state of proteins deposited on solid dry films. Thin, amorphous protein films were fabricated with Apoazurin, alcohol dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase and glutamate dehydrogenase the protein being incorporated into a DEAE-dextran matrix and deposited on quartz slides. The results, obtained with appositely constructed instrumentation, demonstrate that thanks to the low background radiation associated with long-lived, delayed emission phosphorescence can be readily detected down to single protein layer matrices and that both spectrum and lifetime are important indicators of the integrity of the protein globular fold. In fact, denaturation of the proteins by guanidinium hydrochloride or heat treatment points out that disruption of the native fold leads to a red shift and broadening of the spectrum with loss of vibronic structure, accompanied to considerably shorter-lived and more heterogeneous decay kinetics. It is also shown that the sensitivity of the phosphorescence lifetime towards the detection of altered, looser conformations of the polypeptide are remarkably enhanced on partial hydration of the sample. PMID- 11419644 TI - Biosensors based on novel peroxidases with improved properties in direct and mediated electron transfer. AB - Native horseradish peroxidase (HRP) on graphite has revealed approximately 50% of the active enzyme molecules to be in direct electron transfer (ET) contact with the electrode surface. Some novel plant peroxidases from tobacco, peanut and sweet potato were kinetically characterised on graphite in order to find promising candidates for biosensor applications and to understand the nature of the direct ET in the case of plant peroxidases. From measurements of the mediated and mediatorless currents of hydrogen peroxide reduction at the peroxidase modified rotating disk electrodes (RDE), it was concluded that the fraction of enzyme molecules in direct ET varies substantially for the different plant peroxidases. It was observed that the anionic peroxidases (from sweet potato and tobacco) demonstrated a higher percentage of molecules in direct ET than the cationic ones (HRP and peanut peroxidase). The peroxidases with a high degree of glycosylation demonstrated a lower percentage of molecules in direct ET. It could, thus, be concluded that glycosylation of the peroxidases hinders direct ET and that a net negative charge on the peroxidase (low pI value) is beneficial for direct ET. Especially noticeable are the values obtained for sweet potato peroxidase (SPP), revealing both a high percentage in direct ET and a high rate constant of direct ET. The peroxidase electrodes were used for determination of hydrogen peroxide in RDE mode (mediatorless). SPP gave the lowest detection limit (40 nM) followed by HRP and peanut peroxidase. PMID- 11419645 TI - Ascorbate oxidase based amperometric biosensor for organophosphorous pesticide monitoring. AB - An amperometric principle based biosensor containing tissues of cucumber, rich in ascorbic acid oxidase, was used for the detection of organophosphorous (OP) pesticide ethyl paraoxon, which inhibits the activity of ascorbic acid oxidase. The optimal concentration of ascorbic acid used as substrate was found to be 5.67 mM. The biosensor response was found to reach steady state within 2 min. A measurable inhibition (> 10%) was obtained with 10 min incubation of the enzyme electrode with different concentrations of the pesticide. There was a linear relationship between the percentage of inhibition of the enzyme substrate reaction and the pesticide (ethyl paraoxon) concentration in the range 1-10 ppm with a regression value 0.9942. PMID- 11419646 TI - Sensitivity deviation: instrumental linearity errors that influence concentration analyses and kinetic evaluation of biomolecular interactions. AB - Many scientific instruments utilise multiple element detectors, e.g. CCD's or photodiode arrays, to monitor the change in a position of an optical pattern. For example. instruments for affinity biosensing based on surface plasmon resonance (SPR) or resonant mirror are equipped with such detectors. An important and desired property of these bioanalytical instruments is that the calculation of the movement or change in shape follows the true change. This is often not the case and it may lead to linearity errors, and to sensitivity errors. The sensitivity is normally defined as the slope of the calibration curve. A new parameter is introduced to account for the linearity errors, the sensitivity deviation, defined as the deviation from the undistorted slope of the calibration curve. The linearity error and the sensitivity deviation are intimately related and the sensitivity deviation may lead to misinterpretation of kinetic data, mass transport limitations and concentration analyses. Because the linearity errors are small (e.g. 10 pg/mm2 of biomolecules on the sensor surface) with regard to the dynamic range (e.g. 30,000 pg/mm2), they can be difficult to discover. However, the linearity errors are often not negligible with regard to a typical response (e.g. 0-100 pg/mm2). and may therefore cause serious problems. A method for detecting linearity errors is outlined. Further on, this paper demonstrates how integral linearity errors of less than 1% can result in a sensitivity deviation of 10%, a value that in our opinion cannot be ignored in biospecific interaction analysis (BIA). It should also be stressed out that this phenomenon also occurs in other instruments using array detectors. PMID- 11419647 TI - Immobilization of pneumococcal polysaccharide vaccine on silicon oxide wafer for an acoustical biosensor. AB - One the most important aspects of a biosensor is related to immobilization and maintenance of specific reference compounds on sensing surfaces. A method for the immobilization of polysaccharides to a silicon oxide surface intended for Surface Acoustical Waves (SAW) sensors is described. Silicon oxide is a hydrophobic inorganic support used for the fabrication of many electronic devices. The pneumococcal polysaccharide (PPS) vaccine is immobilized via Protein A after pre treatment of the surface with hydrochloric acid. The effects of non-specific binding are discussed. The results indicate that the immobilization of PPS via Protein A increases the sensitivity of detecting Streptococcus pneumoniae antibodies in human sera and offers greater reproducibility of response compared with ELISA methods. The principles of this technique are simple and are applicable to the immobilization of many capsular polysaccharides. PMID- 11419648 TI - Miniaturized real-time monitoring system for L-lactate and glucose using microfabricated multi-enzyme sensors. AB - A miniaturized on-line monitoring system for the detection of L-lactate and glucose is presented. The system is based on a microfabricated multi-enzyme silicon sensor chip with flow channels integrated on the chip. The sensors were fabricated in containment technology. They were characterized in test solutions. The cross-talking behaviour was investigated and was found to be practically negligible. The linear measurement ranges of both glucose and lactate sensors were large enough for most practical applications. As a result of the miniaturization the analyte consumption could be reduced to a few nmol min(-1). The system was equipped with a microdialysis probe whose recovery was 45% for lactate and 37% for glucose in test solutions using a flow rate of 3 microl min( 1). Lower flow rates of 0.5 microl min(-1) resulted in recoveries of over 90%. The long-term stability of the system was acceptable. Initial measurements have also been performed in vitro using human blood serum. PMID- 11419649 TI - Real-time electrochemical imaging using an individually addressable multi-channel electrode. AB - We developed a real-time electrochemical imaging method that uses a multiple enzyme-modified microelectrode. The method will enable the investigation of the functions of biological materials and cells. To test its effectiveness, we imaged the two-dimensional concentration distribution for hydrogen peroxide and L glutamate in a standard solution. The multiple electrode consists of an 8 x 8 array of 30 x 30 microm2 carbon micro electrode. Each electrode was connected to a 64-channel potentiostat that could apply a potential to all electrodes at the same time. The multiple electrode was coated with an Os-polyvinylpyridine based polymer (Os-gel) containing horse radish peroxidase (HRP) to detect hydrogen peroxide, which is a very common product of oxidase enzyme. When measuring glutamate, which is a well-known neurotransmitter in the mammalian central nerve system, we modified the electrode with a bilayer of Os-gel-HRP and GluOx. The detection limit of our method was 1 microM and images of the glutamate concentration-distribution changes induced by local injection of glutamate through microcapillary were obtained in real time. PMID- 11419650 TI - Simultaneous monitoring of glucose and lactate by an interference and cross-talk free dual electrode amperometric biosensor based on electropolymerized thin films. AB - An interference and cross-talk free dual electrode amperometric biosensor integrated with a microdialysis sampling system is described, for simultaneous monitoring of glucose and lactate by flow injection analysis. The biosensor is based on a conventional thin layer flow-through cell equipped with a Pt dual electrode (parallel configuration). Each Pt disk was modified by a composite bilayer consisting of an electrosynthesised overoxidized polypyrrole (PPYox) anti interference membrane covered by an enzyme entrapping gel, obtained by glutaraldehyde co-crosslinking of glucose oxidase or lactate oxidase with bovine serum albumin. The advantages of covalent immobilization techniques were coupled with the excellent interference-rejection capabilities of PPYox. Ascorbate, cysteine, urate and paracetamol produced lactate or glucose bias in the low micromolar range; their responses were, however, completely suppressed when the sample was injected through the microdialysis unit. Under these operational conditions the flow injection responses for glucose and lactate were linear up to 100 and 20 mM with typical sensitivities of 9.9 (+/- 0.1) and 7.2 (+/- 0.1) nA/mM. respectively. The shelf-lifetime of the biosensor was at least 2 months. The potential of the described biosensor was demonstrated by the simultaneous determination of lactate and glucose in untreated tomato juice samples; results were in good agreement with those of a reference method. PMID- 11419651 TI - Enzyme biosensor for studying therapeutics of Alzheimer's disease. AB - An electrochemical method for the investigation and comparison of anti-Alzheimer medications that is based on the inhibition of the acetylcholinesterase is presented. The developed amperometric biosensor determines the in-vitro inhibition of the acetylcholinesterase that is co-immobilized with choline oxidase on the working electrode surface of a three-electrode system using gel entrapment. The sensor has been applied to determine the IC50 values of two known and one newly developed Alzheimer remedy. A simultaneous measurement with the photometric standard method shows the applicability of our method for fast drug screening. PMID- 11419652 TI - pH strategy for deep hypothermic circulatory arrest in adults. PMID- 11419653 TI - A cardiopulmonary bypass score system to assess quality of perfusion performance. AB - During cardiopulmonary bypass, the perfusionist maintains physiological parameters laid down in protocols; this is his or her performance capability. In order to assess his or her performance we need to be able to analyse these physiological parameters objectively. We defined six parameters, pH, BE, PaCO2, PaO2, ACT and oesophageal temperature and gave them ideal values of 7.40+/-0.05, 0.0+/-2.5 mmol/l, 39.0+/-3.0 mmHg, 150+/-50 mmHg, 540+/-60 s and 37.2+/-0.2 degrees C, respectively. We established ranges and a score system: +/- one standard deviation of the mean for a score of zero; between +/- one and two standard deviations for a score of one; and greater than +/- two standard deviations for a score of two. We captured and analysed the most outlying value, with respect to known normal values, for each parameter recorded on the pump sheet. This was performed for 100 consecutive patients. Mean +/- standard deviation (medians) values for pH, BE, PaCO2, PaO2, ACT and oesophageal temperature were 7.41+/-0.07 (7.41), -1.85+/-2.37 mmol/l (-1.85 mmol/l), 34.6+/ 5.42 mmHg (34.0 mmHg), 320+/-96.2 mmHg (317 mmHg), 558+/-164 s (503 s) and 37.3+/ 0.5 degrees C (37.4 degrees C), respectively. We then analysed what percentage of our 100 patients fell within each score range for each of the six parameters. This is an efficient means in analysing whether the perfusionist abides by the protocols, what quality is supplied to the patient, does he or she react when he or she is faced with parameters that are out of range and finally advocating in line blood gas monitoring. This is another step towards our goal of total quality management. PMID- 11419654 TI - An update on perfusion safety: does the type of perfusion practice affect the rate of incidents related to cardiopulmonary bypass? AB - Cardiopulmonary bypass (CPB) techniques vary among adult and pediatric patients undergoing cardiac surgery. This may result in a differential conduct of CPB between various aged patients. The present study reports on perfusion incidents occurring in hospitals using extracorporeal circulation. An 80 question survey was mailed to chief perfusionists at all 1030 US cardiac surgical centers. Respondents were asked to report on device use and incidents occurring during a 2 year period from July 1996 to June 1998. Five hundred and twenty-four completed surveys were returned with the age of surgical patients operated on at each hospital defined as either an adult (n=407), pediatric (n=17), or combined-adult and pediatric (n=100). Centrifugal pumps were used as the primary systemic pumps in 54% of adult, 12% of pediatric, and 36% of combined centers. In-line blood gas monitoring was used in 76% of all pediatric hospitals, but in only 30% of adult facilities. Incident rates occurred once per every 120.9, 83.9, and 220.2 cases in adult, pediatric, and combined centers, respectively. Mortality rates related to CPB occurred 2.7 times higher in adult and pediatric centers as compared to combined hospitals. Arterial dissection was the number one cause of death in both pediatric and combined hospitals, while coagulation disturbances resulted in the highest mortality for adult procedures. Results of this study show that the lowest incident rates occur at hospitals performing combined adult and pediatric CPB. PMID- 11419655 TI - A genetically engineered human Kunitz protease inhibitor with increased kallikrein inhibition in an ovine model of cardiopulmonary bypass. AB - A recombinant human serine protease inhibitor known as Kunitz protease inhibitor (KPI) wild type has functional similarities to the bovine Kunitz inhibitor, aprotinin, and had shown a potential to reduce bleeding in an ovine model of cardiopulmonary bypass (CPB). The aim of this study was to assess KPI-185, a modification of KPI-wild type that differs from KPI-wild type in two amino acid residues and which enhances anti-kallikrein activity in a further double-blind, randomized study in an ovine model of CPB, and to compare with our previous study of KPI-wild type and aprotinin in the same ovine model. Post-operative drain losses and subjective assessment of wound 'dryness' showed no significant differences between KPI-185 and KPI-wild type, despite the significant enhancement of kallikrein inhibition using KPI-185 seen in serial kallikrein inhibition assays. These preliminary findings support the hypothesis that kallikrein inhibition is not the major mechanism by which Kunitz inhibitors such as aprotinin reduce perioperative bleeding. PMID- 11419656 TI - Light and electron microscopic analyses for ischaemia-reperfusion lung injury in an ovine cardiopulmonary bypass model. AB - An experiment to study the role of contact-activation leukocyte sequestration in the formation of ischaemia-reperfusion injury (I-R injury) was carried out. The study was conducted using light and electron microscopic analyses in an ovine cardiopulmonary bypass (CPB) model using a membrane oxygenator. Five adult sheep were used in the study. The CPB circuitry consisted of a roller pump and a membrane oxygenator. During CPB, flow rates ranged from 50 to 60 ml/kg/min with mild hypothermia. The CPB time was fixed at 120 min. Ten minutes after the start of CPB, total CPB was established. Thereafter, total CPB was performed for 100 min, followed by another 10 min of partial CPB. Lung biopsy specimens for light and electron microscopy were obtained from the upper lobe of the right lung before CPB, 109 min after the start of CPB (just before reperfusion) and 30 min after weaning (after reperfusion). A portion of the lung biopsy specimen was taken for a water content measurement at the same time intervals. For measuring the left and right atrial leukocyte counts, blood samples were taken before thoracotomy, 5 and 109 min after the start of CPB, and 30 and 120 min after weaning. C3a was measured before thoracotomy, 109 minafter the start of CPB, and 30 and 120 min after weaning. Plasma malondialdehyde (MDA) was checked before thoracotomy, 109 min after the start of CPB and 30 min after weaning. On both light and electron microscopic examination, mild to moderate acute lung change was observed after ischaemia-reperfusion. Interstitial oedema, leakage of erythrocytes into the alveolar space and endothelial cell swelling were the main findings. However, few neutrophils were seen. Water content of the lung showed a slight increase after the start of CPB, but there was no statistical significance. Neither significant differences in the transpulmonary gradients of leukocytes nor a significant complement activation, expressed by C3a levels, was observed. The MDA level did not display a significant change related to lung reperfusion despite an increase in MDA after the start of CPB. These findings indicate that I-R injury during CPB may not be from complement-activation leukocyte sequestration, but from another source of oxygen free radicals related to CPB. PMID- 11419657 TI - A recovery model of partial cardiopulmonary bypass in the rat. AB - This study was undertaken to develop a recovery model of cardiopulmonary bypass (CPB) in rats. Twenty male Wistar rats (475-550 g) were anaesthetized, mechanically ventilated and the femoral vessels cannulated. The extracorporeal circulation circuit comprised a roller pump, a venous reservoir and a modified Capiox 308 paediatric membrane oxygenator. Priming consisted of 20 ml of fresh homologous blood and 15 ml of colloid. Anticoagulation was achieved with heparin (500 IU/kg). Blood gas analysis, blood pressure monitoring and survival studies were performed in CPB (n=10) and Sham (n=10) rats. Partial CPB was always easily established and was conducted at a flow rate of 100 ml/kg/min for 90 min Blood gas analysis and blood pressure data did not differ between the two groups. All CPB rats survived and the 3-week follow-up period remained uneventful. The rat model of CPB was easy to perform and was associated with excellent survival. This recovery model should allow us to study the pathophysiological processes underlying post-CPB multiple organ dysfunction. PMID- 11419658 TI - The effect of priming techniques of ultrafiltrators on blood rheology: an in vitro evaluation. AB - The increased interest of using ultrafiltration during cardiopulmonary bypass ICPB) has mandated a re-evaluation of the hematological effects of this blood conservation process. 'Rinse-free' ultrafiltrators can be primed using either crystalloid or blood prior to use. It is unknown whether one priming technique results in superior results in ultrafiltration quality. An in vitro circuit was designed to evaluate the Sorin/COBE HC1400 (n=6), the Lifestream HC70 (n=6), and the Terumo/Sarns HC11 (n=6). All test conditions were conducted at a blood flow rate of 250 ml/min and a transmembrane pressure of 250 mmHg. Samples were drawn and analyzed at four distinct time points for hematocrit, total protein, plasma free hemoglobin, interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNFalpha). The HC11 had significantly greater percent increases in hematocrit under the blood priming protocol (29.2+/-7.9) than either the HC1400 (11.0+/-7.8, p<0.03) or the HC70 (11.9+/-7.8, p<0.04). When crystalloid priming was compared to blood priming, the HC1400 and HC70 produced significant percent increases in hematocrit and total protein levels. The HC1400 devices produced significantly less plasma free hemoglobin when primed with crystalloid rather than blood (43.6+/-38.3 vs 21.3+/-5.6, p<0.01). There were no significant differences between devices or priming techniques for IL-6, IL-8 or TNFalpha levels. In conclusion, the efficiency of the ultrafiltrators was elevated when primed with crystalloid before use. Cytokine levels were relatively unchanged with priming techniques, while plasma free hemoglobin levels were reduced with those devices previously primed with crystalloid. PMID- 11419659 TI - Life and death of a cardiac myocyte: principles of cellular biology. AB - If the future of extracorporeal circulation is to include approaches to enhance localized or widespread distribution of cells, and/or gene transfer for augmentation of cardiac function, it is imperative that we gain an increased understanding of the mechanisms that define the cardiac myocyte phenotype. The purpose of this paper is to review the natural history of the cardiac myocyte. A variety of signals determine the cellular processes that characterize birth, growth, differentiation and death of cardiomyocytes. Examined here are primary aspects of the molecular genetics of growth and development, including signal transduction, protein phosphorylation, the cell division cycle, and transcriptional activation. This review is intended to be an update on insights into molecular aspects of the cell, with emphasis on gene expression during cardiac myogenesis and a discussion of its relevance to the field of extracorporeal circulation. In addition, the current status of research in myogenesis is presented. PMID- 11419660 TI - Veno-venous perfusion-induced systemic hyperthermia: case report with perfusion considerations. AB - Cancer cells are more susceptible to destruction by heat than are their normal counterparts. However, optimization of this hyperthermic susceptibility for selective cancer cell kill has been difficult to define and technically difficult to achieve. A whole-body hyperthermic technique veno-venous perfusion-induced systemic hyperthermia (VV-PISH) was designed in in vitro and in swine experiments to achieve selective hyperthermic cancer cell destruction. In this case report, VV-PISH is studied for its safety and therapeutic efficiency in a Food and Drug Administration (FDA) approved phase-I study, where hyperthermia is used to treat advanced (Stage III B or IV) lung cancer. VV-PISH, utilizing the ThermoChem HT system in an extracorporeal circuit, was used to induce hyperthermia to 42.5 degrees C sustained for 120 min. Cooling returned the body temperature to 37 degrees C. After completion of the treatment, the patient was transferred to the intensive care unit on a ventilator, norepinephrine and diuretics. The patient remained somnolent for 36 h, developed pulmonary congestion requiring an additional 48 h before extubation, was transferred to the intermediate unit on day 4 and discharged in good condition on day 8. He did experience hyperthermia related shrinkage of his lung cancer; however, he succumbed 270 days after this treatment from further progression of this disease. Hyperthermia is not a benign therapy; management techniques have been developed that have ameliorated many of the problems associated with extremely high temperatures, but pathophysiology still exists. Using these techniques, VV-PISH can be safety implemented, albeit not without temporary sequelae and further hospitalization. PMID- 11419661 TI - Neurological injury during cardiopulmonary bypass in the rat. PMID- 11419662 TI - Abdominal aortic aneurysm. AB - Abdominal aortic aneurysm is a chronic dilation of the aorta with a natural history toward enlargement and rupture. Its pathogenesis is believed to be multifactorial and complex. Clinical presentation may be asymptomatic, symptomatic, or as rupture. Elective surgery by open transperitoneal or retroperitoneal approach is the most common repair intervention. However, placing an endoluminal stent graft within the aneurysm is currently being evaluated as an alternative to open repair. Nursing care of the patient with abdominal aortic aneurysm involves intensive care skills as well as a foundation in chronic illness management. This article presents information on pathogenesis, natural history, clinical presentation, surgical interventions, and postoperative complications. PMID- 11419663 TI - Aortic dissection. AB - Aortic dissection is the most common catastrophic condition affecting the aorta. Its clinical presentation is variable and can mimic any number of medical and surgical conditions. The most prevalent symptom is excruciating chest pain. Essential treatment includes rapid initiation of pharmacologic agents to control hypertension. Aortic dissection involving the ascending aorta also necessitates immediate surgical repair. Surgical treatment of aortic dissection involving only the descending aorta generally is reserved for patients with persistent pain, intractable hypertension, or evidence of propagation of the dissection. Nursing management includes blood pressure monitoring, serial physical assessment, and postoperative care, as appropriate. PMID- 11419664 TI - Deep venous thrombosis. AB - Venous thrombosis involving the deep veins is a major US health problem that affects over 2.5 million people annually. The most serious complication of a deep venous thrombosis (DVT) is pulmonary embolism (PE), which is associated with 50,000 to 200,000 deaths each year. DVT and PE are often silent and difficult to detect by clinical examination; however, DVT rarely occurs in the absence of risk factors. This article reviews normal venous anatomy and discusses the etiology of DVT, its clinical manifestations, and diagnosis. Then it reviews treatment of DVT, highlighting the nurse's role. A discussion of DVT prophylaxis based on patient risk follows. PMID- 11419665 TI - Peripheral arterial disease of the lower extremity. AB - Peripheral arterial disease is becoming more prevalent due to the increasing age of the population. To better evaluate and care for patients with this disease, it is essential to understand its pathophysiology and etiology. Understanding the sequelae of events and treatments associated with peripheral arterial disease will help the nurse to provide a comprehensive, outcome-focused plan of care and education plan. Nurses are in a unique position to reduce the risks associated with lower extremity arterial disease. PMID- 11419666 TI - Exercise training and smoking cessation as the cornerstones of managing claudication. AB - Based on research, the classic recommendation to prevent progression of lower extremity peripheral arterial disease (PAD) is "start exercising and stop smoking." Unfortunately, a paramount problem for clinicians is motivating targeted individuals to begin and adhere to an exercise regimen and quit smoking. The purpose of this review is to provide the most current information regarding exercise training and smoking cessation. It is hoped that this article will help health care professionals present accurate information to their patients with PAD, address the difficulties in lifestyle change, and intervene effectively. PMID- 11419667 TI - Renovascular hypertension. AB - This article discusses the pathophysiologic features, diagnosis, treatment, and management of patients with renovascular hypertension. Timely diagnosis is important as the disease can carry a more serious prognosis than essential hypertension and is less amenable to drug treatment. Renovascular hypertension also carries a greater risk of progression to accelerated or malignant hypertension and may result in irreversible ischemic failure of the affected kidney. Various treatments are available including medical management, angioplasty and/or stenting, and revascularization. Regardless of treatment approach, nursing care is an integral part of the patient's management, especially continuing follow-up of blood pressure. PMID- 11419668 TI - Pharmacology department. AB - Traditionally administered unfractionated heparin (UH) is a heterogeneous mixture of polysaccharide chains of varying length. Heparin is now available in new formulations, most notably the low-molecular-weight heparins (LMWHs), which possess pharmacology that is similar to but distinct from UH. Key advantages of LMWHs include improved bioavailability and longer half-life, more predictable anticoagulation that requires less laboratory monitoring, and fewer serious side effects. The article reviews the pharmacology of heparin and discusses the addition of LMWHs to the family of anticoagulant drugs. PMID- 11419669 TI - The hidden impact of antibacterial resistance in respiratory tract infection. Re evaluating current antibiotic therapy. AB - Pharmacokinetic/pharmacodynamic (PK/PD) parameters derived from animal and clinical models of infection are used to predict bacteriological efficacy. Growing evidence from the clinical setting supports the validity of these parameters in guiding antimicrobial therapy. For example, in otitis media and sinusitis, high bacteriological cure rates are obtained when serum concentrations of beta-lactams and macrolides exceed the MIC of the infecting pathogen for at least 40% of the dosing interval. Likewise, the 24-hour AUC/MIC ratio is a good predictor of both bacteriological and clinical efficacy for azithromycin in otitis media and fluoroquinolones in bacterial pneumonia. The value of PK/PD relationships has been recognized by the National Committee for Clinical Laboratory Standards (NCCLS) as another important factor to consider when establishing susceptibility breakpoints. Recent changes to NCCLS breakpoints for oral beta-lactams for Streptococcus pneumoniae reflect this. Also, PK/PD parameters may play a role in predicting the impact of an antibiotic on the development and spread of resistant organisms. In an era of increasing resistance, we should select agents and doses that provide drug concentrations that exceed the magnitude of the PK/PD parameter required both for efficacy and to combat the emergence and spread of bacterial resistance. PMID- 11419670 TI - Have we reached the point of no return? The hidden impact of antibacterial resistance in respiratory tract infection. Introduction. PMID- 11419671 TI - The hidden impact of antibacterial resistance in respiratory tract infection. Steering an appropriate course: principles to guide antibiotic choice. AB - The prevalence and degree of antibacterial resistance in common respiratory pathogens are increasing worldwide. The health impact of resistance is not yet fully understood. However, once the impact of resistance becomes measurable, it may be too late to apply interventions to reduce resistance levels and regain previous quality and cost of care. We should address resistance now, before patient care is irreversibly compromised. The association between antibiotic consumption and the prevalence of resistance is widely assumed. However, evidence suggests that there is a more complex. multifactorial relationship between antibiotic use and resistance. It is also assumed that there is an adaptive fitness cost for bacterial resistance mutations. However, in some cases, bacteria are able to acquire 'compensatory genes' negating any negative impact of resistance mutations. Mathematical modeling indicates that the timescale for the emergence of resistance is typically shorter than the decay time following a decline in antibiotic consumption. Against this background, a general principle is proposed: to maximize patient outcome whilst minimizing the potential for selection and spread of resistance. This may be achieved through the use of agents that fulfill defined pharmacodynamic and pharmacokinetic parameters and elicit rapid eradication of the bacterial population, including emerging resistant mutants, from the site of infection. The choice of agent may not be the same in all regions, as selection will depend on local resistance patterns and disease etiology; however, the application of this principle may help to preserve the benefits of antibiotic therapy. PMID- 11419672 TI - The hidden impact of antibacterial resistance in respiratory tract infection. Clinical failures: the tip of the iceberg? AB - In pneumococcal meningitis, it is well accepted that resistance in Streptococcus pneumoniae compromises clinical outcome. However, the clinical impact of increasing resistance on community-acquired respiratory tract infections (RTIs) is less clear. Bacteriological eradication should be the aim of antimicrobial therapy. The pharmacodynamics (potency and pharmacokinetics) of an antimicrobial agent against the infecting pathogen can be used to predict the potential for bacterial eradication. Surveillance of clinical isolates from community-acquired RTIs shows that, in many countries, there is a trend towards an increasing prevalence of drug-resistant S. pneumoniae. Results from a number of published clinical trials suggest that resistance has not compromised the clinical efficacy of aminopenicillins when used at the correct dose. However, emerging data indicate that resistance is compromising the efficacy of some other routinely used antimicrobials. There are reports of clinical and bacteriological failure with macrolides and fluoroquinolones in patients with community-acquired pneumonia. Recent retrospective analyses and increasing sporadic reports of clinical failure with these agents may be more representative of the true situation. These reports suggest a need to reassess current empirical therapeutic recommendations for the treatment of community-acquired RTIs. PMID- 11419673 TI - Polio eradication, and after... PMID- 11419674 TI - The effect of iron supplementation on visual-evoked potentials in infants with iron-deficiency anemia. AB - Flash visual-evoked potentials were studied in 20 infants with iron-deficiency anemia to determine the effect of iron deficiency on visual function by using visual-evoked potentials in this type of anemia. After iron therapy for 12 weeks, visual-evoked potentials were retested in these otherwise healthy infants. All infants showed an excellent hematological response to iron therapy. Post treatment visual-evoked potential N2 latencies (negative deflections) decreased significantly compared to the pre-treatment values (p < 0.05). These results suggest that iron-deficiency anemia causes subclinical visual impairment, and visual-evoked potentials may be a useful non-invasive means of detecting subtle effects of nutritional deficiencies and monitoring the nutritional status of infants. PMID- 11419675 TI - A comparative study of haematological parameters in children suffering from sickle cell anaemia (SCA) from different regions of Saudi Arabia. AB - Sickle cell anaemia (SCA) occurs frequently in several regions of Saudi Arabia but the haematological and clinical presentations are significantly variable. We investigated 264 Saudi children suffering from SCA originating from different regions of Saudi Arabia. Normal children from the same region were used as controls. Freshly obtained blood samples from patients and controls were used to estimate haematological parameters, red cell indices, Hb A2 and Hb F levels. The results of children from the different regions were separately analysed using the Statistical Analysis System (SAS). Significant variations were seen in the haematological parameters in SCA patients compared to controls in each region. In addition, comparison of patients from different areas also showed wide variations. The highest levels of haemoglobin, red blood cells and haematocrit were in the SCA children from the eastern province, and the lowest levels were from those in the western province. Red cell indices and Hb A2 levels did not differ significantly but HbF levels were significantly higher compared to the control group. Inter-regional differences were seen in the Hb F level. A slight but statistically significant increase was seen in the total haemoglobin with Hb F. This paper compares the haematological parameters of SCA in different regions of Saudi Arabia and shows a significant haematological heterogeneity in SCA in Saudis. PMID- 11419676 TI - Risk factors of childhood asthma: a Sri Lankan study. AB - A case-control study was carried out to evaluate the genetic and environmental risk factors of childhood asthma in a group of Sri Lankan children. Three hundred cases (admitted with symptoms of asthma) and 300 age-matched controls were compared over a period of 23 months commencing in January 1996. Family history of atopy, feeding habits in infancy, bronchiolitis in infancy, passive smoking, exposure to dust and dampness, and exposure to pet animals were studied as risk factors for asthma. The risks associated with social factors were also studied. The risk associated with variables were calculated using the chi-squared test in the bivariate analysis and the forward logistic regression model in the multivariate analysis. Parental asthma, asthma in a sibling and in a relative, parental allergic rhinitis, discontinuation of breastfeeding after 6 months in infancy, bronchiolitis in infancy, living in a dusty environment, and a father with primary education compared to secondary education were independently associated with an increased risk of asthma (p < 0.05). This study reinforces the view that asthma has a multifactorial aetiology. Influence of paternal asthma is more than that of maternal asthma. As a preventive measure continuation of breastfeeding beyond 6 months is important. PMID- 11419677 TI - Growth parameters and endocrine function in relation to echocardiographic parameters in children with ventricular septal defect without heart failure. AB - To determine the effect of ventricular function, size of ventricular septal defect (VSD), and endocrine function on linear growth in children with VSD, we studied 88 children with VSD over a period of 1 year. Growth was assessed by determining the height standard deviation scores (HtSDS) and growth velocity (GV) every 4 months. Two hundred age-matched normal children served as controls for the growth data. Endocrine evaluation was performed in 30 randomly selected children with VSD, and 20 age-matched children with constitutional delay of growth (CSS). Growth hormone (GH) response to clonidine provocation was evaluated and circulating free thyroxine (FT4) and insulin-like growth factor-I (IGF-I) concentrations measured. Echocardiographic evaluation of the different cardiac parameters including shunt size and shunt fraction (Qp/Qs) was performed using a colour-coded echodoppler. The HtSDS, body mass index (BMI), and mid-arm circumference (MAC) of children with VSD were significantly decreased compared to those for the normal control group. The dietary intake evaluated by the recall method, appeared to be adequate in the majority of these children (83/88). IGF-I concentrations were reduced in children with VSD (87.5 +/- 29 ng/ml) versus normal age-matched children (169 +/- 42 ng/ml). Basal and clonidine-stimulated GH concentrations were significantly higher in children with VSD (4.6 +/- 2.1 microg/l and 28.8 +/- 7.9 microg/l respectively) versus controls (17.8 +/- 4.2 microg/l). In these patients (n = 88) the HtSDS was correlated negatively with the size of the shunt (r = -0.793, p < 0.001), shunt fraction (Qp/Qs) (r = -0.76, p < 0.001), pulmonary mean gradient (r = -0.4, p = 0.006), and pulmonary maximum velocity (r = -0.32, p = 0.02). Growth velocity (GV) was correlated negatively with pulmonary maximum gradient (r = -0.3, p = 0.02), pulmonary maximum velocity (r = -0.37, p = 0.007), and pulmonary stroke volume (Qp) (r = -0.345, p = 0.01). The BMI and IGF-I concentrations were correlated significantly with the size of the shunt (r = -0.453, p < 0.01), Qp/Qs (r = -0.432, p < 0.01), HtSDS (r = 0.565, p < 0.01), and BMI (r = 0.435, p < 0.01). It appears that in patients with VSD, the size of the left-to-right shunt and the abnormal hemodynamics in the pulmonary circulation are important factors in the etiology of impaired growth. It is suggested that the hypermetabolic status of these patients compromise nutrition and this decreases IGF-I synthesis with subsequent slowing of linear growth and weight gain. PMID- 11419678 TI - Tuberculosis (presumed congenital) in a neonatal unit in Dar-es-Salaam, Tanzania. AB - This is the first report of congenital tuberculosis from Tanzania. It discusses the problems of diagnosis in a typical neonatal unit in a developing country. Three cases are reported within 1 year. Failure to thrive was the most common symptom. We speculate that congenital tuberculosis is not rare and carries a high mortality. There is need to have a high index of suspicion especially where maternal HIV and tuberculosis are highly prevalent. PMID- 11419679 TI - Specific IgA detection can be used for perinatal diagnosis of HIV in children under protocol ACTG 076. AB - Detection of anti-HIV-1 IgA antibodies using a modified ELISA test for the early diagnosis of perinatally acquired HIV-1 infection in children treated with protocol ACTG 076 was evaluated. A total of 177 sera were obtained from 141 infants between 1 and 12 months of age (46 were treated and 95 were non-treated with protocol ACTG 076) and tested for HIV IgA antibodies by an ELISA test after removal of IgG with recombinant protein G. Infants were classified according to CDC's classification system after a follow-up until 20 months of age. Of the 46 treated children 22 turned out to be infected and in the group of 95 untreated children, 52 were infected. All 81 samples from uninfected children treated or untreated with protocol ACTG 076 were persistently IgA-negative. HIV IgA antibodies were detected in 14 of 25 plasma samples from infected children with treatment, and in 58 of 71 samples in infected children without treatment. Considering that the sensitivity of this test is lower in children younger than 6 months the population of children studied was divided into two groups; those under and those over 6 months of age. No significant differences were observed in the detection of IgA in treated or untreated children in both age groups. The overall specificity of the test was 100 per cent; sensitivity in children older than 6 months was 76.92 per cent in treated children and 93.10 per cent in untreated children. In spite of the small number of samples studied it could be demonstrated that treatment with zidovudine does not affect the detection of IgA antibodies. This is a simple and inexpensive method that could be used for diagnosis of treated and untreated children in developing countries. PMID- 11419680 TI - Ten-year experience with paediatric lymph node tuberculosis in Port Moresby. AB - This is a descriptive study of short-course chemotherapy in children with nodal tuberculosis at Port Moresby General Hospital (PMGH). Between 1 August 1989 and 31 December 1997 5248 children were started on TB treatment. In the retrospective study 427 children were treated for lymph node TB up to 31 December 1996. Of these, 207 definitely completed the treatment and 24 (11.6 per cent) of them were known to have relapsed up to the end of 1997. In the prospective study 179 children with lymph node TB were enrolled between 1 January 1997 and 31 December 1997. Of these, 97 definitely completed the treatment and 10 (10.6 per cent) were known to have relapsed during a follow-up period of between 1 and 2 years. PMID- 11419681 TI - Comparison of chloroquine with artesunate in the treatment of cerebral malaria in Ghanaian children. AB - Despite previously reported chloroquine-resistant forms of PF falciparum in Ghana, chloroquine remains the drug of choice in severe malaria. Artemisinin derivatives have been shown to be effective against chloroquine-resistant strains in other endemic areas. This open randomized study was conducted to compare the efficacy of chloroquine and artesunate in the treatment of childhood cerebral malaria. Out of 82 subjects that fulfilled the inclusion criteria, 36 were randomized to receive chloroquine and 46 to receive artemisinin. Blantyre coma scores, temperature and parasitaemia were monitored. Mortality and neurological deficits were documented. There was no difference in mortality rates (chloroquine, 16.7 per cent; artesunate, 21.7 per cent; p = 0.6), neurological deficit at day 14 (chloroquine, 0 per cent; artesunate, 4.3 per cent; p = 0.3), resolution of fever (p = 0.55), and coma recovery time (p = 0.8), between the two groups. The results suggest that syrup chloroquine and intramuscular/oral artesunate currently give comparable clinical responses in the treatment of cerebral malaria in Ghana. Possible reasons for this are discussed, and suggestions are made for future antimalarial drug policy. PMID- 11419682 TI - A comparison of brain, core and skin temperature in children with complicated and uncomplicated malaria. AB - A prospective study was carried out in which brain, core and skin temperatures were studied in children with cerebral malaria (n = 23), uncomplicated malaria (n = 12) and normal children (n = 9) using the zero heat flow method. Patients with cerebral or uncomplicated malaria were admitted to the paediatric wards (mean age, 6 years 8 months +/- 2 years 8 months). Normal children, children of the investigators, of the same age group, served as controls. Parasitaemia levels were similar in the cerebral and uncomplicated malaria cases. Higher brain than core temperatures would have been expected in cerebral malaria but not in uncomplicated malaria but this was not the case in this study. There was no statistical difference in brain, core and skin temperature between cerebral and uncomplicated malaria patients. However, there was a highly significant difference between normal children and cerebral and uncomplicated malaria patients. Brain temperature was 0.02-0.2 degrees C below core temperature in all the groups with larger differences during the febrile period. Mean differences of brain minus core, brain minus skin and core minus skin between the two groups of patients were not statistically significant. There was no correlation between temperature and the level of coma or parasitaemia for cerebral and uncomplicated malaria patients. There was a positive correlation between brain and core temperature in both groups of patients during the febrile phase. Brain temperature remained lower than core temperature in cerebral and uncomplicated malaria as in normal children. Normal thermoregulation appears to be maintained in cerebral malaria. PMID- 11419683 TI - Comparative clinical trial of mebendazole and metronidazole in giardiasis of children. AB - Giardiasis and soil-transmitted helminthosis are prevalent in many parts of the world. Application of a single drug for treatment of both giardiasis and soil transmitted helminthosis is highly desirable. One hundred children of both sexes, ranging from 7 to 12 years old with Giardia intestinalis cysts or trophozoites in their stool samples, were randomly separated into two groups of 50 individuals each. Each group received either mebendazole tablets 200 mg thrice daily for 5 days or metronidazole tablet 15 mg/kg/day in three divided doses, for 7 days. The results were evaluated by microscopic examination of stools on two successive occasions: 1 week and 2 weeks after treatment. Each occasion consisted of three stool examination on 3 successive days. The cure rate of mebendazole-treated children was 43 out of 50 (86 per cent), and for metronidazole it was 45 out of 50 (90 per cent) with no statistical difference between the two groups. No side effects were observed in the mebendazole-treated group, whereas nausea, anorexia and metallic taste were observed in 4.9, 6 and 24 per cent of metronidazole treated children, respectively. In conclusion, the use of mebendazole, which is less toxic with no side-effects, is available and effective on Giardia intestinalis as well as intestinal nematodes, is recommended especially in the areas where giardiasis and intestinal nematodiasis are prevalent and overlap. PMID- 11419684 TI - Mother's working status and nutritional status of children under the age of 5 in urban low-income community, Surabaya, Indonesia. AB - A cross-sectional anthropometrics survey was carried out in a low-income community of Surabaya, Indonesia, to examine the association between mother's employment and the child's nutritional status for identifying a group at risk of having malnourished children. Subjects were 274 children under the age of 5. The children of non-working mothers had significantly higher height-for-age z-score (HAZ) (p < 0.05) than those of working mothers. When mother's work was divided into 'formal' and 'informal', HAZ and weight-for-age z-score (WAZ) of children of the informal worker's group were significantly lower than those of the non working mother and the formal worker's group (p < 0.05). Mother's education and income of the formal worker's group were significantly higher than those of the informal worker's group (p < 0.01). Our study identified a group at risk of malnutrition, i.e., children of mothers working in informal sectors. The programs to provide childcare for working mothers should target this particular group of poor households. PMID- 11419685 TI - Evaluation of serum cholesterol and triglyceride levels in 1-6-year-old Saudi children. AB - Estimations of cholesterol and triglyceride in serum are frequently requested tests due to the close association between elevated levels of these parameters and the risk of arteriosclerosis later leading to cardiovascular disease. Since lipid levels in children show considerable variations in different populations, this study was conducted with the aim of investigating levels of cholesterol and triglycerides in Saudi children less than 6 years old. The study group comprised 582 children with ages ranging from 1 to 6 years, randomly selected during a household screening programme. Fasting blood was used for the estimation of cholesterol and triglyceride using an autoanalyser. The overall range for cholesterol was 2.1-5.7 mmol/l and for triglyceride it was 0.1-1.84 mmol/l. The children were separated into five further groups depending on age, and the levels of cholesterol and triglycerides were obtained in each age group. Using published guidelines for cholesterol and triglyceride levels, to estimate 'borderline' and 'high risk' for arteriosclerosis and coronary artery disease, the prevalence of both risk groups were calculated in Saudi children. A total of 6.87 per cent of children fell in the borderline risk and 1.55 per cent in the high-risk group using cholesterol levels, while 1.89 per cent fell in the borderline-risk group and 1.2 per cent in the high-risk group using triglyceride levels. This paper presents the lipid values and discusses the need for lipid awareness programmes in the country. PMID- 11419686 TI - Urinary free deoxypyridinoline assessment in recognition of rickets. PMID- 11419687 TI - Re-emergence of visceral leischmaniasis: important diagnostic features. PMID- 11419688 TI - Prevalence of epilepsy in 3637 children of primary school age in the province of Malatya, Turkey. PMID- 11419689 TI - The Human Genome Project. PMID- 11419690 TI - Ironing out disease: inherited disorders of iron homeostasis. AB - Iron presents us with a paradox. Without it, cells simply cannot survive because iron is an essential cofactor for many enzymes in critical biochemical pathways. However, when iron is present in excess, it can be highly cytotoxic due to its propensity to catalyze the formation of reactive oxygen radicals. To cater for this dual nature, cells and organisms have developed elaborate mechanisms for regulating iron intake and efflux. When these mechanisms are disrupted, as is the case in a number of inherited disorders of iron metabolism, the pathological consequences can be severe. Many of these disorders are characterized by iron overload and include relatively common diseases such as hereditary hemochromatosis, rare abnormalities of plasma protein synthesis (atransferrinemia and aceruloplasminemia), and the neuromuscular disease Friedreich ataxia. The few described inherited anemias in humans have yet to yield to molecular dissection, but the investigation of several rodent anemias has proved highly rewarding. This review will provide a summary of some of these disorders and describe how their analysis has provided important new insights into iron trafficking pathways and their regulation. PMID- 11419691 TI - Internally located signal peptides direct hepatitis C virus polyprotein processing in the ER membrane. AB - An endoplasmic reticulum (ER) signal peptide is an amino acid sequence motif that directs the translocation of nascent polypeptides to the lumen of ER membrane. Most of known ER signal peptides are either N-terminal cleavable or internally uncleavable. In the structural protein region of hepatitis C virus (HCV) polyprotein, however, four internally located cleavable signal peptides are arranged in a tandem array. The published experimental results indicated that the nascent HCV polyprotein is processed in the ER membrane by host signal peptidase(s) to the respective viral proteins. Here we propose that the four ER signal peptides lead the nascent HCV polyprotein to ER membrane, and the four internally located cleavable signal peptides are the sole determinant for the compartment localization of the matured viral proteins. After cleavage at the C terminus, the signal peptides retain at the C-terminus of mature proteins, and serve as ER membrane anchors. The signal peptide directed polyprotein processing in the ER membrane preludes the virion assembly and budding from the ER membrane. This unique processing may be a general mechanism adopted by many types of virus for virion assembly and replication. The revelation of signal peptidase involved in HCV polyprotein processing presents a novel drug target to suppress HCV viral replication for the much needed HCV therapy. PMID- 11419692 TI - Anti-alpha2 integrin antibody induces secretion and activation of 72-kDa progelatinase by human fibroblasts. AB - We examined the effects of the extracellular matrix proteins fibronectin, laminin, and matrigel on 72-kDa progelatinase secretion by human lip fibroblasts (KD) and skin fibroblasts (MRC9 and HSF). By gelatin zymography, 1.7 +/- 0.2 fold, 1.7 +/- 0.2-fold, and 1.8 +/- 0.3-fold increases were observed in 72-kDa progelatinase secretion from KD cells treated with fibronectin, laminin, and matrigel, respectively. Laminin and matrigel, but not fibronectin, stimulated 72 kDa progelatinase secretion from HSF cells. Fibronectin, laminin, and matrigel did not stimulate 72-kDa progelatinase secretion by MRC9 cells. Anti-alpha2 integrin antibody-stimulated 72-kDa progelatinase secretion and induced the 62 kDa activated form of 72-kDa progelatinase by KD cells. Activated p42 MAP kinase (MAPK) expression was down-regulated by anti-alpha2 integrin antibody. Anti alpha2 integrin antibody stimulated 72-kDa progelatinase secretion by HSF cells without inducing the 62-kDa activated form. The data suggest that interaction between fibroblasts and extracellular matrix components via alpha2 integrin plays an important role in regulating secretion and activation of 72-kDa gelatinase and that down-regulation of activated p42 MAPK may be involved in the 72-kDa progelatinase activation mechanism. PMID- 11419693 TI - Contribution of the residue Glu9, Glu46, and Phe49 to the biological activity of insulin-like growth factor-1. AB - Three recombinant single-chain insulin mutants, [B10Glu]Ins/IGF-1(C), [A5Glu]PIP, and [A8Tyr]PIP were prepared by means of protein engineering. In the three mutants, the residue at position 10 of insulin B chain and positions 5 and 8 of insulin A chain were substituted by the corresponding residue Glu9, Glu46, and an aromatic residue at 49 of IGF-1, respectively. The growth-promoting activity of [B18Glu]Ins/IGF-1(C) is 58-fold higher than insulin and of [A5Glu]PIP, and [A8Tyr]PIP is 2.5- and 4.7-fold higher than PIP, respectively, measured using a mouse mammary tumor cell line GR2H6 in vitro. The results show that the residues Glu9, Glu46, and Phe49, especially the Glu9, play an important role in the growth promoting function of IGF-1. Based on the results, we suggest that the insulin could be used as a scaffold to investigate the contributions of residues of IGF-1 to its biological function. PMID- 11419694 TI - Thiol-oxidizing agent diamide and acidic pH enhance lipid peroxidation of rat heart mitochondria and cardiolipin-cytochrome c complex. AB - In this study, we investigated lipid peroxidation in rat heart mitochondria hydrolyzed by phospholipase A2 (PLA2) and lipid peroxidation in a mitochondrial mimetic lipid peroxidation system, where phospholipids such as cardiolipin (CL) and cytochrome c (Cyt c) were first mixed together and then PLA2 and calcium chloride were added to the mixture (CL-Cyt c-PLA2 system). Production of hydroperoxy and hydroxy compounds of linoleic acid (LA) in the mixture was measured by high performance liquid chromatography. The ratio of the total amount of hydroperoxy and hydroxy compounds of LA to that of LA was calculated as an index for lipid peroxidation (1000 x mol/mol). The index for lipid peroxidation in the rat heart mitochondria hydrolyzed by PLA2 at the physiological pH of 7.4 was 22.8 +/- 2.2 (mean +/- SD, n = 4) and that at the acidic pH of 6.7 was 41.8 +/- 2.0. In the presence of the thiol (SH)-oxidizing agent diamide, the index was 47.0 +/- 2.6 (pH 7.4). In the CL-Cyt c-PLA2 system, lipid peroxidation seemed to be due to three mechanisms: (1) oxidation of the LA (nonreleased form) constituent of CL by Cyt c (oxidation of CL by Cyt c); (2) oxidation of free LA, released from CL, involving the oxidation of CL by Cyt c (free LA oxidation by the CL-Cyt c complex); and (3) oxidation of free LA, released from CL, by Cyt c and calcium ions (LA-Cyt c-Ca system). The lipid peroxidation of the CL-Cyt c PLA2 system was also enhanced by the addition of diamide and by an acidic pH of 6.7. The fact that the SH-oxidizing agent enhanced the lipid peroxidation in the CL-Cyt c-PLA2 system suggested that SH groups in the hemoprotein played an inhibitory role in lipid peroxidation in the system. PMID- 11419695 TI - In vivo ESR study on hepatic reduction of a nitroxide radical after administration of glucose in rats. AB - The in vivo reducing ability of a nitroxide radical, 4-hydroxyl-2,2,6,6 tetramethylpiperidin-1-oxyl (TEMPOL), in the liver of rats was estimated by using an electron spin resonance spectrometer equipped with a surface-coil-type resonator following administration of glucose. Both oral and intravenous administration of glucose significantly enhanced the reduction capacity. In vitro studies show that the reduction site of TEMPOL in the hepatic homogenate is located in the mitochondrial respiratory chain and microsomal electron transport system. These findings indicate that the enhancement of activity of these systems caused by the glucose administration prompts reduction of TEMPOL in the liver. PMID- 11419696 TI - Bradykinin induces protein kinase C-dependent activation of phospholipase D in A 431 cells. AB - The protein kinase C involvement in bradykinin (BK)-induced phospholipase D (PLD) activation in A-431 cells was examined. Treatment of cells with BK induced the rapid activation of intracellular PLD activity. The PLD activation induced by BK was blocked by pretreatment of A-431 cells with staurosporine, or by prolonged treatment with phorbol-12-myristate-13-acetate (PMA). PKC inhibitors Ro-31-8220 and bisindolylmaleimide I, showed the same inhibitory effects on the BK stimulated increase of PLD activity, indicating a role of PKC in this activation process. Similar results were observed in PMA-induced PLD activation. In contrast, PKC down-regulation or PKC inhibitors had no obvious effect on the PLD activation stimulated by epidermal growth factor (EGF). Furthermore, rottlerin and Go 6976, the PKC inhibitors specific for PKC-delta, -alpha and -betaI, respectively, markedly inhibited the PLD activity stimulated by BK. These results indicated that PKC, at least PKC-delta and Ca2+-dependent PKC-alpha or -betaI, plays an important role in BK-induced but not EGF-induced PLD activation in A-431 cells. PMID- 11419697 TI - Frog oocytes: a living test tube for studies on metabolic regulation. AB - This review is intended to illustrate how live frog oocytes may be advantageously used to address the study of some problems of in vivo glucose metabolism. Glucose microinjected into the cells is preferentially committed to glycogen synthesis. We present evidence showing that both the direct and indirect pathways for polysaccharide deposition are operative in oocytes. A small amount of the injected glucose (<5%) is released as labeled CO2 mainly through the pentose-P pathway. Coinjection of NADP+ and glucose significantly stimulates 14CO2 production, half-maximal stimulation being obtained at 0.13 mM. Finally, we show the use of frog oocytes to measure in vivo the control coefficient of hexokinase on glycogen synthesis and the pentose-P pathway. A value of 0.7 was found for the control coefficient of hexokinase on glycogen synthesis, while the enzyme has no control at all over the pentose-P pathway. Therefore, the frog oocyte may be used as a living test tube for the study of almost any metabolic process of interest. PMID- 11419698 TI - Antioxidant enzyme activity and oxidative stress in bovine oocyte in vitro maturation. AB - Reactive oxygen species (ROS) production is a normal process of cell metabolism. In vitro environments usually increase cell production of ROS, which has been implicated as a main cause of cell damage. Nevertheless, the role of ROS in oocyte in vitro maturation (IVM) is controversial. In most cells, enzymatic antioxidant systems can attenuate the effect of oxidative stress by scavenging ROS. The aim of this work was to determine whether: (1) standard conditions of bovine oocyte IVM are responsible for oxidative stress; (2) cumulus cells participate in protection against oxidative stress of the oocyte; and (3) enzymatic antioxidant activity is present in oocytes and cumulus cells. Cumulus oocyte complexes (COCs) were matured in TCM-199 + 10% steer serum for 24 h at 39 degrees C in 5% CO2:95% humidified air. Oxidative stress was determined by the 2',7'-dichlorofluorescein diacetate assay. Superoxide dismutase (SOD), glutathione peroxidase, and catalase activities were measured spectrophotometrically. Under standard conditions of in vitro maturation, there was no increase in ROS production per COC (P > 0.05), but ROS level per cumulus cell diminished. There was no modification in ROS levels in oocytes matured in the presence versus the absence of their surrounding cumulus cells ( P > 0.05). To the best of our knowledge, the presence of SOD, glutathione peroxidase and catalase activities were detected in oocytes and cumulus cells for the first time. Enzymatic units were lower in denuded oocytes with respect to cumulus (P < 0.05), accounting for 37% for SOD, 25% for glutathione peroxidase, and 11% for catalase of the total COC units. Specific enzyme activity diminished in cumulus cells (P > 0.05) and increased in oocytes due to maturation (P > 0.05). The presence of activity of an enzymatic antioxidant system in the bovine oocyte would regulate in part ROS levels during IVM. Oocytes could be capable of controlling the increase in ROS because of the presence of their own enzymatic antioxidant system, SOD having the highest specific activity with respect to cumulus cells. PMID- 11419699 TI - How to spoil your whole day. PMID- 11419700 TI - Optimization of electrophoresis for the identification of low molecular mass allergens in hazelnuts. AB - Conventional electrophoresis techniques used to identify food allergens are insufficient to separate low molecular mass proteins and peptides. In this paper we performed three different methods which provided an extended resolving power for small proteins. Applying the improved techniques, we were able to separate hazelnut proteins into distinct bands below 10 kDa. PMID- 11419701 TI - Incidence of anaphylaxis in the emergency department of a general hospital in Milan. AB - OBJECTIVE: To evaluate incidence and causes of anaphylactic reactions in the emergency room (E.R.) of a general hospital in Milan during a 2-year period. METHODS: We retrospectively studied the computerized records of patients discharged from an E.R. with a diagnosis of anaphylactic reaction. Anaphylaxis was established on the presence of at least two cutaneous, respiratory, gastrointestinal or cardiovascular system symptoms. RESULTS: During 1997 and 1998, out of 38 685 patients referred to the E.R., 13 had severe anaphylaxis with loss of consciousness (LOC) and 127 had anaphylactic symptoms, without LOC. Of the 13 patients with LOC, a possible cause was identified in 12 (five foods, six drugs, one hair dye). In the other 127 patients anaphylaxis was related to foods in 49 cases (38.5%), drugs in 44 (34.6%), unknown causes in 29 (22.8%), hymenoptera stings in two (1.5%), and other causes in three (2.3%). CONCLUSION: The incidence of anaphylactic reactions was 0.4% and mainly affected females and atopic subjects. Foods, particularly fruits and vegetables, appeared to be the most important cause; other important causes were non steroidal antiinflammatory drugs and beta-lactam antibiotics. PMID- 11419702 TI - Bioanalytical characterization of proteins. AB - Allergens from the view of a protein chemist are quite normal proteins, not to distinguish from non allergenic proteins. The first task is therefore to recognize and identify the proteins responsible for the allergenic reaction. This is usually only possible if the allergenic structure is conserved during the purification procedures. For a detailed analysis of the allergenic protein modern protein chemical methods for characterization, identification, determination of posttranslational modifications and epitope characterization have to be applied. Such techniques are briefly described in this article. PMID- 11419703 TI - Determination of the primary structure of two lipid transfer proteins from apricot (Prunus armeniaca). AB - It has been recently demonstrated that the major allergen of apricot is a protein of molecular mass (Mr) 9000 belonging to the family of Lipid Transfer Protein. The aim of this study was the determination of the primary structure of apricot LTP by micro-sequencing and mass spectrometric analyses. Apricot LTP is a 91 amino acids protein like peach and almond LTPs with a sequence identity of 91% and 94%, respectively. Like for the peach LTP, out of the 25 amino acids forming the inner surface of the tunnel-like hydrophobic cavity in maize ns-LTP, 16 are identical and 7 similar in the apricot LTP, supporting the hypothesis of a similar function. PMID- 11419704 TI - Immunological identification and characterization of individual food allergens. AB - Food allergies of type-I-allergy are immunoglobulin E (IgE) mediated and caused by certain proteins or glycoproteins, which are called food allergens. An analytical marker of allergens is the IgE-reactivity to these substances. Normally food allergens are minor components in allergenic source material, which consist of a huge number of chemical different substances. Thus allergen extraction, separation and immunological detection methods are described which identify and characterize individual food allergens by a minimum of manipulation. Favoured separation methods of allergenic extracts are electrophoretic ones allowing the combination of highly resolved protein separations with immunological detection methods subsumed by the term immunoblotting. These techniques are a useful basis to characterize allergens by chemical methods. Once the primary protein structure of a food allergen is established, the way is cleared for the identification of epitopes. Epitopes are immunological detectable parts of a protein or glycoprotein generating the interface between chemical structure and immune-system. The nature of epitopes may differ, for instance, can be conformational, continuous, or built up by glycoconjugates, which determine the stability of food allergens, especially in the case of food processing. Progress in identification and characterization of food allergens will improve diagnostics and therapy of food allergy. PMID- 11419705 TI - Investigations on the carbohydrate moieties of glycoprotein allergens. AB - Many allergens are glycoproteins and their carbohydrate structure can contribute to the IgE reactivity. Therefore it is of great interest to study the carbohydrate structures of these particular antigens. Here, we present an overview of methods combining basic procedures in glycochemistry with various applications of electrophoresis that allow investigating single allergens in crude extracts. Various allergen extracts, e.g. from tomato, grass pollen and bacteria were analysed and the suitability of the tests are discussed. PMID- 11419706 TI - Grid-immunoblotting: a fast and simple technique to test multiple allergens with small amounts of antibody for cross-reactivity. AB - Grid-immunoblotting is a procedure that allows the simultaneous testing of up to 20 different antibodies such as monoclonal antibody-containing hybridoma supernatants or human sera for specific antibodies to up to 20 different antigens or allergens on a single sheet of nitrocellulose membrane. Since only 150 to 200 microl of antibody-containing solution are required this technique is uniquely suited to test growing hybridomas and small amounts of sera (e.g. mouse and children's sera). Compared to a standard ELISA, approximately ten times less antibody is needed to obtain the same information. PMID- 11419707 TI - Characterization and identification of allergen epitopes: recombinant peptide libraries and synthetic, overlapping peptides. AB - For the understanding of the relationship between protein structure and allergenicity, it is important to identify allergenic epitopes. Two methods to characterize primarily linear epitopes are compared using the major allergen from brown shrimp (Penaeus aztecus), Pen a 1, as an example. A recombinant peptide library was constructed and synthetic, overlapping peptides, spanning the entire Pen a 1 molecule, were synthesized and tested for specific IgE reactivity. Both methods identified IgE-binding of Pen a 1, however, the SPOTs procedure resulted in the identification of more epitopes of the major shrimp allergen Pen a 1 than the usage of the recombinant peptide library. For detection of specific IgE antibodies, the usage of 125I-labeled detection antibody seems to be superior over enzyme-labeled anti IgE antibodies. The regeneration of SPOTs membranes is possible, but it is prudent to test regenerated membranes for residual activity. If a given food allergen contains significant linear epitopes, which seems to be true for stable major allergens such as those of peanut and shrimp the SPOTs system may be more advantageous than the use of recombinant peptides libraries. However, if allergens are studied that contain more conformational epitopes, recombinant peptide libraries may help to identify the relevant epitopes. It has to be emphasized that no system for epitope identification will detect all epitopes and that the relevance of identified epitopes has to be confirmed with other methods such as inhibition studies, crystallographic analysis or the immunological evaluation of modified whole allergens. PMID- 11419708 TI - Nuclear magnetic resonance studies of allergens. PMID- 11419709 TI - X-ray structure analysis of food allergens. PMID- 11419710 TI - Structure and function of proteins involved in milk allergies. AB - Allergy to milk proteins has been defined as any adverse reaction mediated by immunological mechanisms to one or several of proteins found in milk. The milk allergy has been classified according to the onset of symptoms as immediate or delayed type. The milk allergy seems to be manifested by three major proteins found in milk: alpha-lactalbumin, beta-lactoglobulin and caseins. The structural comparison of allergenic sites in alpha-lactalbumin and beta-lactoglobulin with the structure of lactoferrin has clearly shown that yet another major milk protein lactoferrin also possesses allergenic sites and thus may qualify to be an allergen. The heat treatment of milk proteins considerably reduces their allergenicity. PMID- 11419711 TI - Structure and properties of ovalbumin. AB - Ovalbumin is a protein of unknown function found in large quantities in avian egg white. Surprisingly, ovalbumin belongs to the serpin family although it lacks any protease inhibitory activity. We review here what is known about the amino acid sequence, post-translational modifications and tertiary structure of ovalbumin. The properties of ovalbumin are discussed in relation to their possible functional significance. These include reasons for failure of ovalbumin to undergo a typical serpin conformational change involving the reactive centre loop, which explains why ovalbumin is not a protease inhibitor, and also the natural conversion of ovalbumin to the more stable "S" form. PMID- 11419712 TI - Seasonal variation in food allergy to apple. AB - The aim of the study was to investigate the possibility of a seasonal variation in reactivity to apples in 27 birch pollen allergic patients. Before and during the birch pollen season 1998, the patients were subjected to double-blind, placebo-controlled food challenges (DBPCFCs) with grated fresh Golden Delicious apple followed by an open food challenge with whole fresh apple. The clinical reactions elicited during the challenges were evaluated both by the patients and the investigators. Moreover, the skin reactivity and the in vitro reactivity to apple were evaluated by skin prick test (SPT), leukocyte histamine release (HR), measurement of specific IgE, and immunoblotting experiments. The sensitivity of the DBPCFC, when compared with the result of the open challenge, was 0.74 (14/19) before the season and 0.80 (16/20) during the season. None of the patients reacted to the blinded challenge without a subsequent reaction to the open challenge. One placebo reaction was registered both before and in season, but not in the same patient. The patient scores of the first positive challenges, and the maximal scores of each combined blinded and open challenge session, were significantly increased during the pollen season (P<0.05). The scores of the open challenge were significantly higher than the scores of the DBPCFC both before the season and during the in-season challenges (P<0.05). Specific IgE against Golden Delicious increased during season (P<0.05), while neither SPT, HR, nor immunoblotting experiments could confirm an increase in reactivity. In conclusion, the results of the oral challenge tests indicated an increase in clinical reactivity to apples during the birch pollen season in birch pollen allergic individuals. PMID- 11419713 TI - Search for the determinants of allergenicity in proteins of the lipocalin family. AB - Three different lines of analysis have been applied to approach the problem of the allergenicity of certain proteins: biological functions, molecular structures and immunological properties. It is immediately obvious that these three are interdependent. The lipocalin family of proteins includes a significant number of allergens. A considerable amount of data is already available of lipocalins and some insights about allergenic determinants can now be presented. However, more information on the molecular structures and immunological parameters of lipocalin allergens is required. PMID- 11419714 TI - Stability of food allergens and allergenicity of processed foods. AB - The allergenicity of food could be altered by several processing procedures. For various foods of animal and plant origin the available literature on this alteration is described. Investigations on hidden allergens in food products are also dealt with. PMID- 11419715 TI - Detection of trace amounts of hidden allergens: hazelnut and almond proteins in chocolate. AB - Many patients with immediate type allergy to tree pollen also suffer from intolerance to hazelnuts and almonds. Since rather low levels of hazelnut and almond proteins can provoke an allergic reaction in sensitized individuals, an immunoblot technique has been developed for the detection of potentially allergenic hazelnut and almond proteins in chocolate. Initially, IgE binding hazelnut and almond proteins were detected by immunoprobing with allergic patients' sera. For routine analysis, patients' sera were substituted with polyclonal rabbit antisera, and sensitivity was enhanced by the use of a chemiluminescent detection method. This technique allowed the detection of less than 0.5 mg of hazelnut or almond proteins per 100 g of chocolate (= 5 ppm). It was applied for routine screening purposes in product quality control as well as for optimization of cleaning steps of filling facilities to minimize cross contamination during production. PMID- 11419716 TI - Determination of the allergenicity of various hazelnut products by immunoblotting and enzyme allergosorbent test inhibition. AB - Although allergic reactions to hazelnuts are common especially in Europe, there are only a few investigations with regard to the influence of processing on the IgE-binding potency of hazelnut proteins. In this study the allergenicity of different hazelnut products, such as chocolate, nougat products, croquant or cookies, was examined by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting and enzyme allergosorbent test (EAST) inhibition experiments using sera of 17 hazelnut-allergic individuals. In only a few cases did the immunoblotting experiments yield positive results as regards the allergenicity of the investigated products. By means of EAST inhibition a residual IgE-binding potency could be detected in almost all of the product extracts. Therefore hazelnuts are a potential hazard to allergic people even as an ingredient of processed foods. PMID- 11419717 TI - Determination of the IgE-binding activity of soy lecithin and refined and non refined soybean oils. AB - In the present study refined and non-refined soybean oils as well as soy lecithins were investigated for residual allergenicity and compared with extracts from native soybeans. By means of immunoblotting and EAST inhibition experiments no IgE-binding activity was detectable in refined soybean oils, which is probably due to thermal treatment during the refining. The investigated non-refined oils and soy lecithins showed a residual IgE-binding activity. In addition in the lecithin extracts a new IgE-binding structure with a molecular mass of approximately 16 kDa was detectable. PMID- 11419718 TI - Recombinant food allergens. AB - Allergenic (glyco)proteins are the elicitors of food allergies and can cause acute severe hypersensitivity reactions. Recombinant food allergens are available in standardised quantity and constant quality. Therefore, they offer new perspectives to overcome current difficulties in the diagnosis, treatment and investigation of food allergies. This review summarises the expression strategies and characteristics of more than 40 recombinant food allergens that have been produced until today. Their IgE-binding properties are compared to those of their natural counterparts, in addition their application as diagnostic tools, the generation of hypoallergenic recombinant isoforms and mutants for therapeutic purposes, the determination of epitopes and cross-reactive structures are described. PMID- 11419719 TI - Pyr c 1, the major allergen from pear (Pyrus communis), is a new member of the Bet v 1 allergen family. AB - Pear is known as an allergenic food involved in the 'oral allergy syndrome' which affects a high percentage of patients allergic to birch pollen. The aim of this study was to clone the major allergen of this fruit, to express it as bacterial recombinant protein and to study its allergenic properties in relation to homologous proteins and natural allergen extracts. The coding region of the cDNA was obtained by a PCR strategy, cloned, and the allergen was expressed as His-Tag fusion protein. The fusion peptide was removed by treatment with cyanogen bromide. Purified non-fusion protein was subjected to allergenicity testing by the enzyme allergosorbent test (EAST), Western blotting, competitive inhibition assays, and basophil histamine release. The deduced protein sequence shared a high degree of identity with other major allergens from fruits, nuts, vegetables, and pollen, and with a family of PR-10 pathogenesis related proteins. The recombinant (r) protein was recognised by specific IgE from sera of all pear allergic patients (n = 16) investigated in this study. Hence, the allergen was classified as a major allergen and named Pyr c 1. The IgE binding characteristics of rPyr c 1 appeared to be similar to the natural pear protein, as was demonstrated by EAST-inhibition and Western blot-inhibition experiments. Moreover, the biological activity of rPyr c 1 was equal to that of pear extract, as indicated by basophil histamine release in two patients allergic to pears. The related major allergens Bet v 1 from birch pollen and Mal d 1 from apple inhibited to a high degree the binding of IgE to Pyr c 1, whereas Api g 1 from celery, also belonging to this family, had little inhibitory effects, indicating epitope differences between Bet v 1-related food allergens. Unlimited amounts of pure rPyr c 1 are now available for studies on the structure and epitopes of pollen-related food allergens. Moreover, the allergen may serve as stable and standardised diagnostic material. PMID- 11419720 TI - Patient-tailored cloning of allergens by phage display: peanut (Arachis hypogaea) profilin, a food allergen derived from a rare mRNA. AB - A peanut cDNA phage surface display library was constructed and screened for the presence of IgE-binding proteins. We used a serum from a peanut-sensitized individual with a low specific IgE level to peanut extract and suffering from mild symptoms after peanut ingestion. A total of 10(11) cDNA clones were screened by affinity selection towards serum IgE immobilized to solid-phase supports. After five rounds of selective enrichment, sequence determination of 25 inserts derived from different clones revealed presence of a single cDNA species. The cDNA-encoded gene product, formally termed Ara h 5, shows up to 80% amino acid sequence identity to the well-known plant allergen profilin, a 14 kD protein present only in low amount in peanut extracts. Immunoblot analysis of fifty sera from individuals sensitized to peanut showed that 16% had mounted a detectable IgE response to the newly identified peanut profilin. High-level expression as non-fusion protein in BL21 (DE3) was carried under control of the inducible T7 promoter. Peanut profilin was purified by affinity chromatography on poly-(L proline)-Sepharose and yielded 30 mg l(-1) culture of highly pure recombinant allergen. In spite of the high level of up to 80% amino acid identity to other plant profilins, inhibition experiments with recombinant profilins of peanut, cherry, pear, celery and birch revealed marked differences regarding their IgE binding capacity. PMID- 11419721 TI - Adverse reactions to foods. AB - Allergic reactions to foods represent a prominent, actual and increasing problem in clinical medicine. Symptoms of food allergy comprise skin reactions (urticaria, angioedema, eczema) respiratory (bronchoconstriction, rhinitis), gastrointestinal (cramping, diarrhea) and cardiovascular symptoms with the maximal manifestation of anaphylactic shock. They can be elicited by minute amounts of allergens. The diagnosis of food allergy is done by history, skin test, in vitro allergy diagnosis and--if necessary--oral provocation tests, if possible placebo-controlled. Avoidance of respective allergens for the allergic patient, however, is often complicated or impossible due to deficits in declaration regulations in many countries. Increasing numbers of cases including fatalities, due to inadvertent intake of food allergens are reported. It is therefore necessary to improve declaration laws and develop methods for allergen detection in foods. Allergens can be detected by serological methods (enzyme immunoassays, in vitro basophil histamine release or in vivo skin test procedures in sensitized individuals). The problem of diagnosis of food allergy is further complicated by cross-reactivity between allergens in foods and aeroallergens (pollen, animal epithelia, latex etc.). Elicitors of pseudo-allergic reactions with similar clinical symptomatology comprise low-molecular-mass chemicals (preservatives, colorings, flavor substances etc.). For some of them (e.g. sulfites) detection assays are available. In some patients classic allergic contact eczema can be elicited systemically after oral intake of low-molecular mass contact allergens such as nickel sulfate or flavorings such as vanillin in foods. The role of xenobiotic components in foods (e.g. pesticides) is not known at the moment. In order to improve the situation of the food allergic patient, research programs to elucidate the pathophysiology and improve allergen detection strategies have to be implemented together with reinforced declaration regulations on a quantitative basis. PMID- 11419722 TI - Molecular basis of allergic cross-reactivity between group 1 major allergens from birch and apple. AB - Patients allergic to birch pollen often also react with fruits and vegetables, such as apple. The major cause of cross-reactivity between birch and apple is biochemical and immunological similarity between the major allergens, Bet v 1 and Mal d 1, as demonstrated by serological and cellular immunoassays. In addition, birch pollen-specific therapeutic allergy vaccination has been shown to improve allergic symptoms caused by oral ingestion of apple. Detailed analysis of molecular surface areas based on the crystal structure of Bet v 1, and primary sequence alignment, identify potential epitopes for cross-reactive antibodies. Two or more conserved patches are identified when comparing Bet v 1 and Mal d 1, thus providing a molecular model for serological cross-reactivity involving more than one IgE-binding epitope. A minimum of two epitopes would be necessary for cross-linking of receptor bound IgE in functional histamine release assays and skin test. Individual amino acid substitutions, as occurring in isoallergenic variation, may, however, have a dramatic effect on epitope integrity if critical residues are affected. Thus, one area large enough to accommodate antibody binding epitopes shared by all known Mal d 1 isoallergens and variants is identified, as well as areas shared by Bet v 1 and individual Mal d 1 isoallergens or variants. The occurrence of limited epitope coincidence between Bet v 1 and Mal d 1 is in agreement with the observation that some, but not all, birch pollen allergic patients react with apple, and that the epitope repertoire recognised by the IgE of the individual patients determines the degree of cross reactivity. PMID- 11419723 TI - Cross-reactivity within the profilin panallergen family investigated by comparison of recombinant profilins from pear (Pyr c 4), cherry (Pru av 4) and celery (Api g 4) with birch pollen profilin Bet v 2. AB - Profilin is a panallergen which is recognised by IgE from about 20% of birch pollen- and plant food-allergic patients. Little is known about epitope diversity among these homologous proteins, and about the correlation between IgE-cross reactivity and allergenic reactivity. Plant food profilins from pear (Pyr c 4) and cherry (Pru av 4) were cloned by polymerase chain reaction and produced in Escherichia coli BL21. The profilins were purified as non-fusion proteins by affinity chromatography on poly-(L-proline)-Sepharose and characterized by immunoblotting, IgE-inhibition experiments and histamine release assays. The coding regions of the cDNA of pear and cherry profilin were identified as a 393 bp open reading frame. The deduced amino acid sequences showed high identities with birch pollen profilin Bet v 2 (76-83%) and other allergenic plant profilins. Pyr c 4 and Pru av 4 were investigated for their immunological properties in comparison with profilins from celery (Api g 4) and birch pollen (Bet v 2). Fourty-three of 49 patients (88%), preselected for an IgE-reactivity with Bet v 2 showed specific IgE-antibodies to the recombinant pear protein, 92% of the sera were positive with the recombinant cherry allergen and 80% of the sera were reactive with the celery protein. Inhibition experiments showed a strong cross reactivity of IgE with profilins from plant food and birch pollen. However, IgE binding profiles also indicated the presence of epitope differences among related profilins. All investigated profilins, Pyr c 4, Pru av 4, Api g 4 and Bet v 2, presented almost identical allergenic properties in cellular mediator release tests. Therefore, cross-reactivities between related profilins may explain pollen related allergy to food in a minority of patients. The nucleotide sequences reported have been submitted to the Genbank database under accession numbers AF129424 (Pyr c 4) and AF129425 (Pru av 4). PMID- 11419724 TI - Genetic modification and plant food allergens: risks and benefits. AB - Plant genetic engineering has the potential to both introduce new allergenic proteins into foods and remove established allergens. A number of allergenic plant proteins have been characterized, showing that many are related to proteins which have potentially valuable properties for use in nutritional enhancement, food processing and crop protection. It is therefore important to monitor the allergenic potential of proteins used for plant genetic engineering and major biotechnology companies have established systems for this. Current technology allows gene expression to be down-regulated using antisense or co-suppression and future developments may allow targeted gene mutation or gene replacement. However, the application of this technology may be limited at least in the short term by the presence of multiple allergens and their contribution to food processing or other properties. Furthermore, the long-term stability of these systems needs to be established as reversion could have serious consequences. PMID- 11419725 TI - What establishes a protein as an allergen? AB - There is little known about the factors that determine the allergenicity of food proteins. Apparently, the ability of a food protein to induce an allergic response requires its presence in substantial amounts in the food supply, its durability during food processing, and its resistance to digestion in the gastrointestinal tract. In addition to the mode and degree of exposure, structural characteristics appear to play an important role for the capacity of a protein to modulate the immune response towards allergic reactions. Until now, however, there has been no indication for common structural characteristics of linear T cell or linear IgE (B cell) epitopes and the knowledge of structural characteristics of conformational IgE binding sites is very limited. Experimental data point only to certain surface areas of allergenic proteins which are important for IgE binding. Therefore, it is not possible to suggest any structural motif or conformational sequence pattern common to all allergenic proteins. Furthermore, glycosylation appears not to be a common critical determinant of allergenicity since food allergens comprise both glycoproteins and nonglycosylated proteins. Based on the few published three-dimensional structures of allergenic proteins including food proteins, one unifying feature of allergens appears to be their spherical shape. The three-dimensional structures of many more allergens have to be determined, however, to allow for a better understanding of the molecular basis of allergenicity. Most recently, new ideas have been introduced as to why certain biochemical or biologic functions such as enzymatic activities may predispose a protein to become an allergen. Proteolytically active allergens have been demonstrated to irritate the human mucosal surface, to enhance their own transmucosal uptake, and to augment IgE production. Therefore, the functional activity of some allergens may play a role among other factors in the process of sensitization and allergic responses. PMID- 11419726 TI - Food legislation and the protection of allergic and hypersensitive persons: an overview. AB - So far there are worldwide no legal instruments in protecting people against adverse allergic reactions to the consumption of foods. Instruments, which generally were developed for health protection in food legislation, are not suitable to regulate the protection of allergic persons because they automatically would exclude all protein-containing foods from commercial market. The only approach to an effective protection is to indicate the presence of adverse effect causing agents or ingredients on the label. This preventive instrument was developed by the Codex Alimentarius and led to an open "hit list" of ten Major Serious Allergens, which have to be labeled. The hit list principle was adopted by the EU-Commission as a draft for an EC-Directive. Unsolved problems are exeptions from the labeling requirement for refined oils and a minimum limit of adverse effect causing agents which do not require labeling. PMID- 11419727 TI - In vivo and in vitro techniques to determine the biological activity of food allergens. AB - Methods for determination of the biological activity of food allergens comprise both determination of the allergenic potency, i.e. the capability to elicit an allergic reaction in an already sensitized individual, and the allergenic potential, i.e. the risk for sensitizing a hitherto non-allergic individual. Several methods are discussed for determination of potency including the double blinded placebo-controlled food challenge, skin testing, in vitro effector cell assays such as basophil histamine release, and IgE-based techniques such as RAST and RAST inhibition. No reliable methods have yet been developed which can predict the allergenic potential of a food or a food allergen. The progress in the areas of stability studies and animal models for food allergy are discussed. PMID- 11419728 TI - Standardization of food allergen extracts for skin prick test. AB - The aim of the study was to standardize and evaluate technically optimized food allergen extracts for use in skin prick test (SPT). The standardization procedure comprised 36 allergic histories in 32 food allergic patients with 21 healthy, non atopic individuals serving as controls. The patients had a history of allergic symptoms upon ingestion of either cow's milk (n=3), hen's egg (n=9), wheat (n=4), hazelnut (n=14) or cod (n=6). They also had specific IgE in serum to the food in question and a positive SPT with a fresh preparation of the food. The diagnosis had been confirmed by a double-blind, placebo-controlled food challenge, except for the hazelnut-allergic patients. The controls were subjected to an open food challenge with all the foods to ensure tolerance. The standardization was performed by means of titrated SPT in accordance with the guidelines on biological standardization from the Nordic Council on Medicine. Regression analysis of the skin wheal areas was performed for each patient and the median protein concentration of allergen preparation (median Ch10) eliciting a wheal area of the same size as histamine 10 mg/ml was calculated. The median Ch10 was 0.56 mg/ml for milk, 0.88 mg/ml for egg, 5.4 mg/ml for wheat, 2.1 mg/ml for hazelnut and 0.017 mg/ml for the cod extract. The sensitivity of the median Ch10 estimated from the SPT data was 1 for milk, 0.98 for egg, 1 for wheat, 1 for hazelnut and 0.87 for the cod extract. The allergenic activity of the hazelnut extract was further investigated by leukocyte histamine release (HR) and immunoblotting experiments using sera from 27 hazelnut allergic patients. The clinical sensitivity of the optimized hazelnut extract evaluated by HR was 0.78 compared to 0.30 for a commercially available hazelnut extract (Soluprick). Immunoblotting results showed a stronger IgE binding capacity and additional IgE binding bands of the optimized hazelnut extract compared with the Soluprick extract. PMID- 11419729 TI - Isolation of food allergens. AB - The identification of food allergens is a priority in the management of food allergy, because of the need to obtain standardized extracts and pure allergens for diagnosis and therapy. It is thus important to develop methods for purification of allergenic molecules in order to study their biological and immunological characteristics. Protocols for protein extraction from foods and for allergen purification are reviewed in this paper. We report published methods for extraction of allergens from either animal and vegetable foods and detailed purification methodologies including ion-exchange, gel filtration and reversed phase chromatography of well known allergens. PMID- 11419730 TI - The major allergen of sesame seeds (Sesamum indicum) is a 2S albumin. AB - BACKGROUND: Allergic reactions induced by ingestion of foods containing sesame seeds are a well recognized cause of severe food-induced anaphylaxis. OBJECTIVE: This study aimed to identify and characterize the clinically most important major allergen of sesame seeds. METHODS: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and IgE immunoblotting were performed on sera of 10 patients selected for severe and documented allergic reaction after eating food containing sesame. The major allergen was purified by gel filtration and characterized by isoelectric point (pI), glycosylation and amino acid sequencing. RESULTS: All the patients had positive IgE antibodies and skin prick tests (SPTs) to sesame. The major, clinically most important allergen was a protein with molecular mass of about 9000. It was not glycosylated, the amino acid sequence showed it was a 2S albumin with a pI of 7.3; the small and the large subunits, forming the whole protein, showed pI values of 6.5 and 6.0. PMID- 11419731 TI - Characterization of the major allergen of plum as a lipid transfer protein. AB - BACKGROUND: Allergy to Prunoideae fruit (plum, peach, cherry and apricot) is one of the most frequent food allergies in southern Europe. All these fruits cross react in vivo and in vitro, as they share their major allergen, a 9 kD lipid transfer protein (LTP). OBJECTIVE: The aim of the study was the identification and molecular characterization of the major allergen of plum. METHODS: The IgE pattern of reactivity to plums was investigated by SDS-PAGE and immunoblotting with the sera of 23 patients. The identified major allergen was purified by HPLC, using a cationic-exchange column followed by gel-filtration. Further characterization was achieved by periodic-Schiff stain, isoelectrofocusing and N terminal amino acid sequencing. RESULTS AND CONCLUSIONS: The major allergen of plum is a 9 kD lipid transfer protein, not glycosylated and with a basic character (pI>9), highly homologous to the major allergen of peach. PMID- 11419732 TI - Improved methods for determining the concentration of 6-thioguanine nucleotides and 6-methylmercaptopurine nucleotides in blood. AB - The conversion of the cytotoxic and immunosuppressive 6-mercaptopurine (6MP) to the active 6-thioguanine nucleotides (6TGN) is necessary for clinical efficacy of 6MP and its prodrug azathioprine. Another metabolite, 6-methylmercaptopurine nucleotide (6MMPN), is formed via a competing pathway by thiopurine methyl transferase. The concentrations of 6TGN and 6MMPN are measured in washed erythrocytes as a surrogate to the intracellular levels of these metabolites in the target tissues. Analysis of 6TGN and 6MMPN in multi-center clinical studies is more complicated because of the requirement to wash erythrocytes. In this investigation, we found no differences in the concentrations of 6TGN and 6MMPN in blood versus washed erythrocytes in samples obtained from patients taking therapeutic doses of oral 6MP or azathioprine for inflammatory bowel disease. We concluded that whole blood could be used for the analysis of these analytes, thus saving sample preparation time. We also found that the erythrocyte 6TGN concentration in blood at ambient temperature declined 2-4% per day, a loss that can be avoided by shipping blood samples frozen. The loss of 6TGN in blood stored at approximately -80 degrees C was 1% after 1 week and 12% after 24 weeks, indicating the analyte was moderately stable. 6MMPN in blood did not significantly change after 24 weeks of storage at approximately -80 degrees C. In addition, the sensitivity of the 6TGN assay was improved by modifying the HPLC conditions, which made the method more suitable for quantifying low levels of 6TGN in human intestinal biopsy samples and blood. PMID- 11419733 TI - Synthesis, high-performance liquid chromatography-nuclear magnetic resonance characterization and pharmacokinetics in mice of CD271 glucuronide. AB - Retinoic acid-glucuronides are known as retinoids with activity in acne therapy, limited placental transfer and reduced retinoid adverse effects. We synthesized the glucuronide of a novel retinoid, CD271 (adapalene), used for the treatment of moderate acne. The synthesis product ("CD271 glucuronide", CD271G) was purified by preparative HPLC. It undergoes in aqueous solution, like other glucuronides, rapid acyl-migration of the bound aglycone leading to position isomers. Thus characterization of purified CD271G could be only achieved by HPLC-NMR coupling. A subfraction ("CD271GB") consisting essentially of 2'- and 3'-CD271G was used for pharmacokinetic studies. After a single subcutaneous injection at a dosage of 30 mg/kg the substance showed considerable uptake and metabolism to CD271 indicating that CD271GB could serve as a prodrug for CD271. PMID- 11419734 TI - Multi-residue extraction-purification procedure for corticosteroids in biological samples for efficient control of their misuse in livestock production. AB - A fast and efficient multi-residue extraction-purification procedure was developed for 12 corticosteroids in biological matrices (hair, urine and meat), in order to control their illegal use as growth promoters in cattle. Detection and identification of the analytes were achieved using a previously described LC MS-MS method based on negative electrospray ionisation and a triple quadrupole analyser. The presented procedures included acid (hair) or enzymatic (urine and meat) hydrolysis, C18 reversed-phase SPE, Na2CO3 liquid-liquid clean-up and SiOH normal-phase SPE. The detection limits of the developed methods were between 2.9 and 9.3 pg/mg (ppb) for hair samples and in the 40 - 70 pg/g (ppt) range for the urine or meat samples. The acid hydrolysis used for corticosteroid extraction in hair was optimised using an experimental design and response surface methodology. Achieved performances were linked to a physico-chemical approach based on the corticosteroids specific C17 side-chain. This original multi-residue and multi matrices analytical methodology will be used for further metabolism studies. PMID- 11419735 TI - Determination of polyamines in human saliva by high-performance liquid chromatography with fluorescence detection. AB - A high-performance liquid chromatographic method for the determination of polyamines (spermine, spermidine and putrescine) in human saliva was developed. This method is based on pre-column derivatization with o-phthaldialdehyde (OPA). The derivatives were separated on a Nucleosil ODS column (250x4.6 mm I.D.; 5 microm). The gradient elution was performed with two mobile phases A (water) and B (methanol) at a flow rate of 0.8 ml/min. The column eluate was monitored by fluorescence detection (excitation, 360 nm; emission, 510 nm). The within- and between-assay coefficients of variation for all the compounds were below 5%. The detection limits for spermine, spermidine and putrescine were 0.04, 0.05 and 0.06 nmol/ml, respectively. The recovery was greater than 90%. Our analytical technique requires neither preliminary extraction with an organic solvent, nor long multi-step procedures. For saliva samples, this is a simple, rapid and highly reproducible method that can be easily applied to the routine determination of salivary polyamines, whose levels increase early in several pathological conditions. PMID- 11419736 TI - Determination of free D-aspartic acid, D-serine and D-alanine in the brain of mutant mice lacking D-amino acid oxidase activity. AB - A simple and precise method for the simultaneous determination of free D-aspartic acid, D-serine and D-alanine in mouse brain tissues was established, using a reversed-phase HPLC system with widely used pre-column derivatizing reagents, o phthaldialdehyde and N-t-butyloxycarbonyl-L-cysteine. With the present method, the contents of these three D-amino acids in hippocampus, hypothalamus, pituitary gland, pineal gland and medulla oblongata as well as cerebrum and cerebellum of mutant mice lacking D-amino-acid oxidase activity were determined and compared with those obtained for control mice. In both mice, extremely high contents of D serine were observed in forebrain (100-400 nmol/g wet tissue), and the contents were small in pituitary and pineal glands. While, D-serine contents in cerebellum and medulla oblongata of mutant mice were about ten times higher than those in control mice. In contrast, D-alanine contents in mutant mice were higher than those in control mice in all brain regions and serum. PMID- 11419737 TI - High-performance liquid chromatographic assay for the simultaneous determination of lansoprazole enantiomers and metabolites in human liver microsomes. AB - In this study, a simple, sensitive and enantioselective HPLC method was developed for the simultaneous determination of lansoprazole enantiomers: a proton pump inhibitor, and its major metabolites: 5-hydroxylansoprazole and lansoprazole sulfone in human liver microsomes. After extraction from the microsomal incubation mixture with a diethyl etherdichloromethane (7:3, v/v) mixture, analytes were measured by reversed-phase HPLC on a Chiralcel OD-R column. Detection was made using an ultraviolet absorbance detector set at a wavelength of 285 nm. The mobile phase consisted of a methanol-water (75:25, v/v) mixture. At a flow-rate of 0.5 ml/min, the total run time was 35 min. The limit of quantification for both lansoprazole enantiomers was 0.25 microM and for the metabolites 0.13 microM. The method is suitable for the analysis of lansoprazole enantiomers and its metabolites from human microsomal liver incubations. PMID- 11419738 TI - Determination of etoposide in human plasma and leukemic cells by high-performance liquid chromatography with electrochemical detection. AB - This paper describes a high-performance liquid chromatographic method with electrochemical detection for the determination of etoposide levels in plasma, total and non-protein bound concentration, and in leukemic cells. The precision for between-runs (n=6) was 7.0, 4.9, and 9.5%, the accuracy was 3.7, 7.1 and 6.3%, and within-runs precision (n=6) was 3.9, 2.9 and 5.1% for total plasma, non protein bound plasma fraction and leukemic cells, respectively. The correlation coefficients (R2) were 1.00 for all calibration curves. These assays have been applied to analyze samples from one patient with acute myelogenous leukemia during 24 h after i.v. infusion of etoposide (100 mg/m2). PMID- 11419739 TI - Simultaneous determination of taxol and its metabolites in microsomal samples by a simple thin-layer chromatography radioactivity assay--inhibitory effect of NK 104, a new inhibitor of HMG-CoA reductase. AB - The inhibitory effect of NK-104, a potent inhibitor of HMG-CoA reductase, on taxol metabolism was examined using radio-TLC. This method is described for in vitro measurement of taxol metabolites as an alternative to the commonly used HPLC assay. After incubation of 14C-taxol with human liver microsomes, the supernatants were developed using a solvent system consisting of toluene-acetone formic acid (60:39:1, v/v) and quantified with a bioimaging analyzer. The described method provides a valuable tool for the simultaneous determination of unchanged taxol and its major metabolites. There was no inhibitory effect of NK 104 on CYP-mediated metabolism of taxol in human liver microsomes. PMID- 11419740 TI - Comparison between liquid chromatography-time of-flight mass spectrometry and selected reaction monitoring liquid chromatography-mass spectrometry for quantitative determination of idoxifene in human plasma. AB - This study compares HPLC electrospray time-of-flight mass spectrometry and selected reaction monitoring (SRM) LC-MS for high throughput quantitative determination of a small molecule drug in biological samples. A high throughput LC-MS method was developed for quantitatative determination of idoxifene in human plasma and the evaluation was accomplished with the cross-validation of the developed LC-MS method between the time-of-flight mass spectrometer, and a triple quadrupole mass spectrometer operated in the SRM mode. A simple one-step semi automated 96-well liquid-liquid extraction procedure was used to prepare 96 samples in approximately 30 min and a rapid gradient was used to shorten the LC run time. Time-of-flight mass spectrometry provides acquisition of full-scan mass spectra and extracted ion current chromatograms, which may be extracted from the total ion current chromatogram for peak area determination. The limit of quantitation for idoxifene in human plasma obtained with the time-of-flight mass spectrometer was 5 ng/ml based on 100-microl aliquots of human plasma, and the linear dynamic range was from 5 ng/ml to 2000 ng/ml. The quantitative LC-MS results from the time-of-flight mass spectrometer demonstrated that precision did not exceed 7.1% and accuracy did not exceed 1.7% with reference to quality control samples at three concentration levels in replicates of six. In contrast, the limit of quantitation for idoxifene in human plasma using a tandem triple quadrupole mass spectrometer was 0.5 ng/ml with a linear dynamic range to 1000 ng/ml. The results from the triple quadrupole instrument show that the precision did not exceed 2.2% and accuracy did not exceed 2.9%. The overall results suggest time-of-flight mass spectrometry may be a viable technique for high throughput bioanalytical work for the quantitative determination of a representative small molecule drug in the low ng/ml range in human plasma. PMID- 11419741 TI - Ultrasensitive assay for three polyphenols (catechin, quercetin and resveratrol) and their conjugates in biological fluids utilizing gas chromatography with mass selective detection. AB - The concentrations of three polyphenols ((+)-catechin, quercetin and trans resveratrol) in blood serum, plasma and urine, as well as whole blood, have been measured after their oral and intragastric administration, respectively, to humans and rats. The method developed for this purpose utilized ethyl acetate extraction of 100 microl samples and their derivatization with bis(trimethylsilyl)trifluoroacetamide (BSTFA) followed by gas-chromatographic analysis on a DB-5 column followed by mass selective detection employing two target ions and one qualifier ion for each compound. Total run time was 17 min with excellent resolution and linearity. The limits of detection (LOD) and quantitation (LOQ) were an order of magnitude less than for any previously published method, being 0.01 microg/l and 0.1 microg/l, respectively, for all compounds. Recovery at 1 microg/l and 10 microg/l was >80% in all instances but one, and was >90% in 50%. Imprecision was acceptable at 0.25 and 1.0 microg/l, concentrations below the LOQ of previous methods. Aglycones released from conjugates after hydrolysis were easily measurable. Optimal conditions for hydrolysis were established. After oral administration of the three polyphenols to humans, their conjugates vastly exceeded the concentrations of the aglycones in both plasma and urine. Concentrations peaked within 0.5-1.0 h in plasma and within 8 h in urine. During the first 24 h, 5.1% of the (+)-catechin and 24.6% of the trans-resveratrol given were recovered in the urine (free plus conjugated). This method can be proposed as the method of choice to assay these polyphenols and their conjugates in biological fluids. PMID- 11419742 TI - Simultaneous determination of the histamine H1-receptor antagonist ebastine and its two metabolites, carebastine and hydroxyebastine, in human plasma using high performance liquid chromatography. AB - Ebastine (CAS 90729-43-4) is an antiallergic agent which selectively and potently blocks histamine H1-receptors in vivo. A simple and sensitive high-performance liquid chromatography (HPLC) method is described for the simultaneous determination of ebastine and its two oxidized metabolites, carebastine (CAS 90729-42-3) and hydroxyebastine (M-OH), in human plasma. After a pretreatment of plasma sample by solid-phase extraction, ebastine and its metabolites were analyzed on an HPLC system with ultraviolet detection at 254 nm. Chromatography was performed on a cyano column (250x4.0 mm I.D.) at 40 degrees C with the mobile phase of acetonitrile-methanol-0.012 M ammonium acetate buffer (20:30:48, v/v/v) at a flow rate of 1.2 ml/min. Accurate determinations were possible over the concentration range of 3-1000 ng/ml for the three compounds using 1 ml plasma samples. The intra- and inter-day assay accuracy of this method were within 100+/ 15% of nominal values and the precision did not exceed 12.4% of relative standard deviation. The lower limits of quantitation were 3 ng/ml for ebastine and its metabolites in human plasma. This method was satisfactorily applied to the determination of ebastine and its two oxidized metabolites in human plasma after oral administration of ebastine. PMID- 11419743 TI - Validation of a simple and sensitive gas chromatographic method for the analysis of tri-n-butyl phosphate from virally inactivated human immunoglobulin. AB - Tri-n-butyl phosphate (TnBP), a solvent used in combination with Triton X-100 to inactivate lipid-enveloped viruses from immunoglobulin purified from human plasma is routinely measured in our laboratories by gas chromatography-flame ionization dectection (GC-FID) after extraction with C-18. We modified our present assay by extracting the analyte into hexane prior to measurement by GC-FID. We also found that the addition of a small volume of ethanol to the organic layer facilitates the extraction process by breaking the resulting emulsion formation caused by the hexane addition. The sample preparation and subsequent assay were fully validated in our laboratory. The process time for each sample is less than 2 min, a 15-fold improvement over solid-phase extraction techniques that were previously used in our laboratories. The recovery of TnBP in immunoglobulin using this newer method approximates 100%. The limit of quantitation (LOQ) was found to be 2 microg/ml or 2 ng per injection. The linear dynamic range of the assay is reported to be from the LOQ up to 50 microg/ml. The method is simple, relatively inexpensive and rapid. In addition, validation of the method demonstrates that it is accurate, precise, rugged and robust as demonstrated by reproducibility between analysts, instruments, laboratories, and columns. Finally, no problems were observed with regard to sample carryover. PMID- 11419744 TI - Determination of monohydroxyethylrutoside in heart tissue by high-performance liquid chromatography with electrochemical detection. AB - 7-Monohydroxyethylrutoside (monoHER) is one of the components of the registered drug Venoruton. It showed a good protection against the cardiotoxic effects of doxorubicin. The analysis of monoHER was developed to study the pharmacokinetic profile of the drug in heart tissue. MonoHER was extracted from heart tissue homogenate with methanol. The supernatant was diluted 1:1 (v/v) with 25 mM phosphate buffer and injected onto a reversed-phase ODS column. The mobile phase consisted of 49% methanol and 51% of an aqueous solution containing 10 mM sodium dihydrogenphosphate (pH 3.4), 10 mM acetic acid and 36 microM EDTA. The retention time of monoHER was about 5.2 min and no endogenous peaks were interfering. The lower limit of quantification was 0.072 nmol g(-1) wet heart tissue. The calibration line was linear up to 24 nmol g(-1). The within-day accuracy and precision of the quality controls (0.12, 1.2 and 12.0 nmol g(-1)) were smaller than 17 and 19%, respectively. The between-day accuracy and precision were better than 6 and 11%, respectively. The recovery of monoHER from heart tissue ranged from 104.1 to 114.3% and was concentration independent. MonoHER was stable in heart tissue when stored at -80 degrees C for 6 months. Repeated injection of monoHER from aliquots of 7.2 nmol g(-1) placed on the sample tray at 4 degrees C for 24 h showed a decrease in the concentration of 30.3%. Analyzing sample duplicates in a mirror image sequence could compensate for the influence of this gradual decrease. The small sample volume allowed one to measure monoHER in the hearts of mice. PMID- 11419745 TI - Non-ideal behaviour of free vanadate on a Superose 12 size-exclusion column. Application to in vivo 48V-labelled rat spleen homogenate. AB - Seven chromatographic columns were evaluated for the recovery of 48V radiolabelled vanadate. Further, the behaviour of vanadate (H2VO4-) was studied on a size-exclusion column Superose 12 as a function of (a) buffer salt molarity, (b) different buffer salts, (c) different buffers and (d) organic solvents added to the buffer. As opposed to the unsatisfactory recovery of V-compounds on other columns, we recovered the vanadium quantitatively. We observed that in most cases vanadate eluted after the total volume of the Superose 12 column. This indicates a non-ideal behaviour of vanadate. However, through this non-ideal behaviour it was possible to separate low-molecular-mass bound (Mr<5000) and unbound vanadium which would not be possible under normal behaviour. A possible explanation for this non-ideal behaviour of vanadium is put forward. The method has been successfully applied for the fractionation of different vanadium species in rat spleen homogenate. PMID- 11419746 TI - Determination of hydrogen sulfide and acid-labile sulfur in animal tissues by gas chromatography and ion chromatography. AB - A sensitive and reliable method was developed for the determination of hydrogen sulfide and acid-labile sulfur (ALS) in animal tissues using gas chromatography with flame photometric detector (GC-FPD) and ion chromatography (IC). Hydrogen sulfide trapped in alkaline solution was determined by GC-FPD as hydrogen sulfide or by IC as sulfate after oxidation with hydrogen peroxide. Sodium sulfide used as a source of hydrogen sulfide was standardized by IC. Fresh rat liver and heart tissues contained 112.2+/-23.0 and 274.1+/-34.6 nmol/g of ALS respectively. Free hydrogen sulfide was not detected. PMID- 11419747 TI - Determination of sulfadoxine in human blood plasma using packed-column supercritical fluid chromatography. AB - A sensitive, rapid, selective and reproducible method has been developed to measure plasma levels of sulfadoxine, 4-Amino-N-(5, 6-dimethoxy-4-pyrimidinyl) benzensulfonamide; in healthy, human volunteers using packed-column super critical fluid chromatography. Omeprazole, 5-methoxy-2-[[(4-methoxy-3, 5-di methyl-2-pyridinyl)methyl]sulfinyl]-1H-benzimidazole; was used as the internal standard (i.s.) at 15.0 microg/ml. The drug and the i.s. were extracted from plasma using dichloromethane. Separation of sulfadoxine and i.s. was done on a Nucleosil (250x4.6 mm) 10 microm, RP-C18 column with 7.4% (v/v) methanol-modified supercritical fluid carbon dioxide (2.5 ml/min) as the mobile phase. The column temperature was 40 degrees C and the outlet pressure was set at 8.83 MPa. The detection was done using a UV-Vis detector set at 265 nm. The limit of quantification was 0.50 microg/ml using 1 ml plasma specimen. The mean extraction recovery of the drug from plasma was found to be 94.9%. The SFC method was directly compared to a published HPLC/UV method. With respect to speed and use of organic solvents SFC was found to be superior; while in all other aspects the results were similar to the published technique. The method has been successfully used to estimate the sulfadoxine levels in healthy human volunteers from 0 to 240 h following an oral dose of 500 mg of sulfadoxine in combination with 25 mg of pyrimethamine. PMID- 11419748 TI - Screening, confirmation and quantification of diuretics in urine for doping control analysis by high-performance liquid chromatography-atmospheric pressure ionisation tandem mass spectrometry. AB - A sensitive, selective, robust and fast method to identify 32 diuretics and masking agents in urine is described. The analytical procedure is reduced to a single XAD extraction step for sample preparation, followed by reversed-phase liquid chromatography in combination with atmospheric pressure ionisation/tandem mass spectrometry. This technique is, after minor modifications, suitable for screening analyses and confirmation of identity as well as quantitation of diuretics. Considerations relating to the stability and metabolism of the compounds are given if relevant for routine screening analyses. PMID- 11419749 TI - Determination of styrene and styrene-7,8-oxide in human blood by gas chromatography-mass spectrometry. AB - Methods of isotope-dilution gas chromatography-mass spectrometry (GC-MS) are described for the determination of styrene and styrene-7,8-oxide (SO) in blood. Styrene and SO were directly measured in pentane extracts of blood from 35 reinforced plastics workers exposed to 4.7-97 ppm styrene. Using positive ion chemical ionization, styrene could be detected at levels greater than 2.5 microg/l blood and SO at levels greater than 0.05 microg/l blood. An alternative method for measurement of SO employed reaction with valine followed by derivatization with pentafluorophenyl isothiocyanate and analysis via negative ion chemical ionization GC-MS-MS (SO detection limit=0.025 microg/l blood). The detection limits for SO by these two methods were 10-20-fold lower than gas chromatographic assays reported earlier, based upon either electron impact MS or flame ionization detection. Excellent agreement between the two SO methods was observed for standard calibration curves while moderate to good agreement was observed among selected reinforced plastics workers (n = 10). Levels of styrene in blood were found to be proportional to the corresponding air exposures to styrene, in line with other published relationships. Although levels of SO in blood, measured by the direct method, were significantly correlated with air levels of either styrene or SO among the reinforced plastics workers, blood concentrations were much lower than previously reported at a given exposure to styrene. The two assays for SO in blood appear to be unbiased and to have sufficient sensitivity and specificity for applications involving workers exposed to styrene and SO during the manufacture of reinforced plastics. PMID- 11419750 TI - Simultaneous determination of reduced and oxidized glutathione in peripheral blood mononuclear cells by liquid chromatography-electrospray mass spectrometry. AB - We developed a sensitive and specific liquid chromatography-electrospray mass spectrometric (HPLC-ESI-MS) assay for the simultaneous determination of reduced and oxidized glutathione (GSH and GSSG) in peripheral blood mononuclear cells (PBMC). Following derivatization with N-ethylmaleimide to prevent GSH auto oxidation, addition of thiosalicylic acid as internal standard, and protein precipitation with cold acetonitrile, the samples were injected into a diol column, eluted with acetonitrile-1% aqueous acetic acid (25:75) and detected by the ESI-MS system. The optimized method exhibited a good detection limit for both analytes (0.01 and 0.05 microM for GSH and GSSG, respectively). Good linearity was reached in the 0.01-20 microM range for GSH and 0.05-20 microM for GSSG. The mean recoveries of GSH and GSSG were 98.5-100.6% and 105.8-111.5%, respectively. The run-to-run repeatability for retention time and peak area was RSD% 0.06 and 1.75 for GSH and 0.18 and 2.50 for GSSG. The optimized method was applied to GSH and GSSG assay in PBMC analyzing 20 healthy individuals. PMID- 11419751 TI - Improved capillary electrophoresis conditions for the separation of kinase substrates by the laser micropipet system. AB - Phosphorylated and nonphosphorylated forms of peptide substrates for protein kinase C (PKC) and calcium-calmodulin activated kinase II (CamKII) were separated by capillary zone electrophoresis. Electrophoresis of the peptide substrates and products in biologic buffer solutions in uncoated capillaries yielded asymmetric analyte peaks with substantial peak tailing. Some of the peptides also exhibited broad peaks with unstable migration times. To improve the electrophoretic separation of the peptides, several strategies were implemented: extensive washing of the capillary with a base, adding betaine to the electrophoretic buffer, and coating the capillaries with polydimethylacrylamide (PDMA). Prolonged rinsing of the capillaries with a base substantially improved the migration time reproducibility and decreased peak tailing. Addition of betaine to the electrophoretic buffer enhanced both the migration time stability as well as the theoretical plate numbers of the peaks. Finally PDMA-coated capillaries brought about significant improvements in the resolving power of the separations. These modifications all utilized an electrophoretic buffer that was compatible with a living biologic cell. Consequently they should be adaptable for the new capillary electrophoresis-based methods to measure kinase activation in single cells. PMID- 11419752 TI - Retention-property relationships of anticonvulsant drugs by biopartitioning micellar chromatography. AB - Epilepsy may be considered as a group of disorders with only one thing in common: the fact that recurrent anomalous electrochemical phenomena appear in the central nervous system. Different classes of drugs are included under the generic term of anticonvulsant drugs. All of them work by decreasing discharge propagation in different ways. Biopartitioning micellar chromatography (BMC) is a mode of reversed-phase liquid chromatography, which can be used as an in vitro system to model the biopartitioning process of drugs when there are no active processes. In this paper, relationships between the BMC retention data of anticonvulsant drugs, their pharmacokinetics (oral absorption, protein binding, volume of distribution, clearance, and renal elimination) and their therapeutic parameters (therapeutic, toxic and comatose-fatal concentration, and LD50) are studied and the predictive ability of models is evaluated. PMID- 11419753 TI - Biological control of leachate from municipal landfills. AB - Landfilling is still a popular way for municipal solid waste (MSW) treatment. Leachate generated from landfills is becoming a great threat to the surroundings as it contains high concentrations of toxic substances. How to control leachate migration and to protect environmental pollution is now a concern for many environmentalists. In this work, eight effective microorganisms (EMs) were isolated from wastewater, sludge and soil samples by enrichment culturing techniques and used for leachate migration control in columns and pilot experiments. The preliminary experiments reveal that the EMs could remove 25% and 40% of chemical oxygen demand (COD) from leachate in fine sand and sabulous clay columns, respectively. An aquifer system was designed to simulate in-situ control for leachate migration with EMs. The EMs were injected into the simulated aquifer and formed a permeable biological barrier. The experimental results show that the barrier removed 95% of COD and approximately 100% inorganic nitrogen, that is, nitrate-N plus nitrite-N plus ammonia-N, from the migrating leachate. CO2 production, redox potential and microbial number were monitored simultaneously in the aquifer during the experiment to assess the EMs' activities and the effect of the bio-barrier. The data indicate that the EMs isolated in this work had high activities and were effective for organic and nitrogenous contaminant removal throughout the experiment. PMID- 11419754 TI - Quantum yield study of the photodegradation of hydrophobic dyes in the presence of acetone sensitizer. AB - The photodegradation of hydrophobic disperse dyes with different chromophores in the presence of acetone (ACE) was investigated. In this study, the photodecay of dyes was carried out in the Rayonet RPR-200 merry-go-round photoreactor, with 253.7 nm monochromatic ultraviolet (UV) lamps. A typical azo disperse dye (CI disperse yellow 7--DY7) and an anthraquinone disperse dye (CI disperse orange- DO11) were used as the probe compounds. The results demonstrate that the addition of acetone increases the solubility of hydrophobic disperse dyes and enhances the photosensitization reaction simultaneously. More than ten times of quantum yield enhancement is observed in the presence of ACE photosensitizer than in water alone. The photodegradation of DY7 and DO11 is dominated by photoreduction, which follows pseudo first-order decay, and the rate constants strongly depend on the solvent system (i.e., ACE/H2O ratios) and the initial pH levels. The decay quantum yields of dyes are normally observed with the increase of the ACE/H2O ratio. The optimum quantum yields of DY7 and DO11 were determined at 0.5 (v/v) and 0.25 (v/v), respectively, in alkaline conditions. A further increase in the ACE/H2O ratio reduces the quantum yields, possibly due to light attenuation by excess acetone. PMID- 11419755 TI - Technology for recovering phosphorus from incinerated wastewater treatment sludge. AB - A bench-top study of phosphorus-recovering technology from ash associated with incinerated wastewater (sewage) treatment sludge was conducted by adding sulfuric acid to ashes for the elution of phosphorus. With the exception of lead, which is insoluble in sulfuric acid, when the pH of the ash fell below 2.0, phosphorus and various heavy metals in the ash were extracted. The study found that, when alkalis were added to adjust the pH of the ash extract to 4.0, phosphorus was subsequently recovered via filtration. Furthermore, when alkalis were added to adjust the pH to 10, the recovery of various heavy metals was observed. In addition, disposal of the remaining solution (wastewater), which consists of a relatively low concentration of salts, is not considered to be a significant issue since it is within wastewater discharge standards and has been found to be useful as an acid-treating substance. PMID- 11419756 TI - Enhanced sludge granulation in upflow anaerobic sludge blanket (UASB) reactors by aluminum chloride. AB - Two upflow anaerobic sludge blanket (UASB) reactors were concurrently operated for 146 days to examine the effects of aluminum chloride (AlCl3) on the sludge granulation process during start-up. Sludge granulation (defined as that over 10% of granules were larger than 2.0 mm) was achieved in the control reactor (R1) in approximate three months. Introduction of Al3+ at a concentration of 300 mg/l reduced the sludge granulation time by approximate one month. Throughout the experiment the AlCl3-added reactor (R2) had a higher biomass concentration, e.g., 13.8 g-MLVSS/l versus 12.8 g-MLVSS/l on Day 146. Granules became visible earlier in R2 compared with R1 (35 days versus 65 days). The average size of granules from R2 was larger than that from R1. The results demonstrated that AlCl3 enhanced the sludge granulation process in the UASB reactors. PMID- 11419757 TI - Efficiency of biological treatment affected by high strength of ammonium-nitrogen in leachate and chemical precipitation of ammonium-nitrogen as pretreatment. AB - Leachate samples with a high strength of ammonium-nitrogen (NH4+-N) were collected from a local landfill site in Hong Kong. Two experiments were carried out to study (1) the inhibition of microbial activity of activated sludge by NH4+ N and (2) the chemical precipitation of NH4+-N from leachate as a preliminary treatment prior to the activated sludge process. The experimental results demonstrated that the efficiency of COD removal decreased from 97.7% to 78.1%, and the dehydrogenase activity of activated sludge decreased from 9.29 to 4.93 microg TF/mg MLSS, respectively, when the NH4+-N concentration increased from 53 to 800 mg/l. The experiment also demonstrated that the NH4+-N in the leachate can be quickly precipitated as MgNH4PO4 x 6H2O after addition of MgCl2 x 6H2O + Na2HPO4 x 12H2O. The NH4+-N concentration was reduced from 5618 to 112 mg/l within 15 min when a molar ratio of Mg2+:NH+:PO4(3-) = 1:1:1 was used. The optimum pH to reach the minimum solubility of MgNH4PO4 x 6H2O was found to be in the range of 8.5-9.0. Attention should be given to the high salinity formed in the treated leachate by using MgCl2 x 6H2O + Na2HPO4 x 12H2O, which may affect microbial activity in the following biological treatment processes. Using two other combinations of chemicals [MgO + 85%H3PO4 and Ca(H2PO4)2 x H2O + MgSO4 x 7H2O] could minimise salinity generation after precipitation, while they were less efficient for NH4+-N removal. PMID- 11419758 TI - Optimising the preparation of activated carbon from digested sewage sludge and coconut husk. AB - Preparation of activated carbon from sewage sludge is a promising way to dispose of sewage sludge as well as to produce a low-cost adsorbent for pollutant removal. This research work aimed to optimise the condition for activated carbon preparation from anaerobically digested sewage sludge with the additive coconut husk. The sewage sludge sample was mixed with the additive coconut husk. The preparation condition variables investigated involved the concentration of the ZnCl2 solutions, heating temperature, dwell time and heating rate in pyrolysis and the mixing ratio of coconut husk to sewage sludge. Surface area, pore size distribution, aqueous phenol adsorption capacity and the production yield of the final products were determined and compared. Experimental results revealed that low concentrations of ZnCl2 solution tended to improve the microporosity of the final product. Heating temperature had a considerable impact on the surface area, pore size distribution and phenol adsorption capacity of the final products, whereas dwell time and heating rate performed comparatively insignificantly. The effect of increasing the mixing ratio of coconut husk to sewage sludge was principally to increase the microporosity of the final products. The activated carbon with the highest BET surface area was produced with the activation of 5 M ZnCl2 solution and, thereafter, pyrolysis at a heating temperature of 500 degrees C for 2 h with a heating rate of 10 degrees C/min. The mixing ratio of 1:4 in terms of coconut husk to sewage sludge based on their dried weights was found to be most cost effective. PMID- 11419759 TI - A comparative study of anaerobically digested and undigested sewage sludges in preparation of activated carbons. AB - Disposal of sewage sludge is an increasingly expensive and environmentally sensitive problem throughout the world. Preparation of activated carbon from sewage sludge offers an attractive re-use alternative to the traditional disposal routes. The objective of this research work was to compare anaerobically digested sewage sludge (DS) and undigested sewage sludge (US) as source materials in the preparation of activated carbons. Prior to the preparation the properties of the two types of sewage sludges were determined and compared. Subsequently the sludge samples were activated with 5 M ZnCl2 solution and thereafter pyrolysed at heating temperature of 650 degrees C for 2 h with the heating rate of 15 degrees C/min under a nitrogen atmosphere. The produced activated carbons were characterised by surface area and porosity analysis, CHN elemental composition and ash contents determination, and aqueous phase phenol adsorption tests. The results indicate that in comparison with the DS, the US had a higher carbon content and lower ash content, and accordingly yielded a better activated carbon with a higher BET surface area, pore volume, carbon content and phenol adsorption capacity. PMID- 11419760 TI - Priority assessment of toxic substances in life cycle assessment. III: Export of potential impact over time and space. AB - Toxicity potentials are scaling factors used in life cycle assessment (LCA) indicating their relative importance in terms of potential toxic impacts. This paper presents the results of an uncertainty assessment of toxicity potentials for 181 substances that were calculated with the global nested multi-media fate, exposure and effects model USES-LCA. The variance in toxicity potentials resulting from choices in the modelling procedure was quantified by means of scenario analysis. A first scenario analysis showed to what extent potential impacts in the relatively short term are obscured by the inclusion of impacts on the very long term. Toxicity potentials representing potential impacts over time horizons of 20, 100 and 500 years were compared with toxicity potentials representing potential impacts over an infinite time horizon. Time horizon dependent differences up to 6.5 orders of magnitude were found for metal toxicity potentials, while for toxicity potentials of organic substances under study, differences remain within 0.5 orders of magnitude. The second scenario analysis addressed to what extent potential impacts on the continental scale are obscured by the inclusion of impacts on the global scale. Exclusion of potential impacts on the global scale changed the toxicity potentials of metals and volatile persistent halogenated organics up to 2.3 orders of magnitude. These scenario analyses also provide the basis for determining exports to future generations and outside the emission area. PMID- 11419761 TI - Environmental and biological monitoring of occupational exposure to organic micropollutants in gasoline. AB - An exposure risk assessment of workers in a refinery production unit was undertaken. Gasoline and its main components were investigated through environmental and biological monitoring. Measured variables were environmental benzene, toluene, pentane and hexane; benzene and toluene in blood and urine; tt MA (metabolite of benzene) in urine. Multivariate statistical analysis of the data showed that worker's exposure to the above substances fell within the limits specified by organisations such as ACGIH. Also, biological values complied with reference values (RV) for non-occupationally-exposed population. Different values of biological variables were determined by separating smokers from non-smokers: smokers had hematic and urinary benzene values significantly higher than non smokers. During a 3-yr sampling, it was possible to identify a significant decrease of benzene in the workplace air and of hematic benzene for non-smokers. The most exposed department, one in which tank-lorries were loaded, needs further investigation and extended monitoring. PMID- 11419762 TI - Luminescence investigations of redox cycling of adriamycin. AB - The light emission from the adriamycin + Co2+ + H2O2 system has been studied. Chemiluminescence, fluorescence and absorption spectra were measured. The fluorescence spectra were time-dependent exhibiting maxima at 555, 590 and 645 nm. The chemiluminescence spectra consist of four bands with maxima at around 460 500, 550-580, 640 and 700 nm. Free radical reaction inhibitors, (1)O2-quenchers and catalase inhibited the light emission indicating that hydroxyl radical, superoxide anion radical and singlet oxygen are generated during the redox cycling of adriamycin. Chemiluminescence studies revealed that adriamycin undergoes chemiexcitation under our experimental conditions. PMID- 11419763 TI - Management, disposal and recycling of waste industrial organic solvents in Hong Kong. AB - An attempt has been made to establish a mass balance of industrial organic solvents in Hong Kong. It is estimated that only a small portion, less than 4%, of all the organic solvents consumed in Hong Kong are collected as waste solvents and properly treated, while the remainder are used either in the formulation of solvent containing products, or are lost to the environment through vapour emissions, leaks and spills, or dumped illegally. It was found that solvent recycling has been a common practice in some industries but the existing level of solvent recycling in Hong Kong is difficult to estimate. About 87.4% of all the waste organic solvents disposed of at the licensed facilities are potentially recyclable although whether they can be recycled in practice depends on many factors. Examples of existing waste organic solvent management and recycling practices from selected industries in Hong Kong are presented. The economic feasibility of current and future potential recycling systems is evaluated for a few selected cases. An integrated waste organic solvent management strategy is proposed to minimize adverse impacts of organic solvents to the environment and human health. PMID- 11419764 TI - Indigenous Australian children: educating for health. PMID- 11419765 TI - How bright is their future? Post-streptococcal glomerulonephritis in Indigenous communities in Australia. PMID- 11419766 TI - Tobacco's uncounted victims. PMID- 11419767 TI - Childhood post-streptococcal glomerulonephritis as a risk factor for chronic renal disease in later life. AB - OBJECTIVE: To test the hypothesis that post-streptococcal glomerulonephritis (PSGN) in childhood is a risk factor for chronic renal disease in later life. DESIGN: Retrospective cohort study. SETTING: A remote Aboriginal community in the "Top End" of the Northern Territory that experienced two epidemics of PSGN in 1980 and 1987, respectively. PARTICIPANTS: 472 people who were aged 2-15 years during either epidemic. They were categorised by clinical features recorded during the epidemics as having clinically defined PSGN (63), "abnormal urine" (haematuria or proteinuria; 86) or controls (323). OUTCOME MEASURES: Urinary albumin to creatinine ratio (ACR), haematuria (by dipstick urinalysis), blood pressure, serum creatinine level, and calculated glomerular filtration rate (GFR) during community screening in 1992-1998. RESULTS: Overt albuminuria (ACR > 34 mg/mmol) was present at follow-up in 13% of the PSGN group, 8% of the abnormal urine group, and 4% of the control group. The odds ratio (OR) for overt albuminuria in those with a history of PSGN compared with the control group, adjusted for age and sex, was 6.1 (95% CI, 2.2-16.9). Haematuria (>trace) was present in 21% of the PSGN group compared with 7% of the control group (adjusted OR, 3.7; 95% CI, 1.8-8.0). There were no significant differences between the groups in blood pressure, serum creatinine level or calculated GFR. CONCLUSION: In this population, a history of PSGN in childhood is a risk factor for albuminuria and haematuria in later life. PMID- 11419768 TI - Improving diabetes care in the primary healthcare setting: a randomised cluster trial in remote Indigenous communities. AB - OBJECTIVE: To evaluate a system for improving diabetes care in remote Indigenous communities. DESIGN: Randomised, unblinded cluster trial over one year (1 March to 29 February 2000). PARTICIPANTS AND SETTING: Primary healthcare staff in 21 primary healthcare centres in the Torres Strait and Northern Peninsula Area (NPA) Health Service District, north Queensland, and 678 people with diabetes, mostly Torres Strait Islanders. INTERVENTION: Diabetes recall system established at eight of the 21 sites, as well as staff training in basic diabetes care, regular phone calls from the project officer, a two-monthly newsletter and a mid-project workshop. MAIN OUTCOME MEASURES: Regular checks of weight, blood pressure, eye and foot care, serum lipid levels and glucose monitoring and control, urinary albumin to creatinine ratio and serum creatinine levels, and administration of recommended vaccines; hospitalisation in the previous 12 months. RESULTS: There was improvement in most measures at most sites, except for blood pressure monitoring and control, and vaccination status. Intervention sites showed greater improvement in most indicators than control sites (combined relative risk [RR], 1.21; 95% CI, 1.03-1.43). The intervention group showed a 32% reduction in hospital admissions for diabetes-related conditions over the study period (P=0.012). At follow-up, patients in intervention sites were 40% less likely to be hospitalised for a diabetes-related condition than those in control sites (RR, 0.60; 95% CI, 0.41-0.86; P=0.007). CONCLUSION: A simple recall system, managed by local healthcare workers and supported by a diabetes outreach service, achieved significant improvements in diabetes care and reduced hospitalisations in a high risk population. PMID- 11419769 TI - Differing influences on Aboriginal and non-Aboriginal neonatal phenotypes: a prospective study. AB - OBJECTIVE: To understand the differences, if any, in major influences on birth phenotype between Aboriginal and non-Aboriginal neonates. DESIGN: Prospective study of a cohort of pregnant Aboriginal women presenting for antenatal care before 20 weeks' gestation (ultrasound proven), and a reference cohort of pregnant non-Aboriginal women. Comparison of the phenotypic and demographic characteristics of the women, their pregnancies and their babies. PARTICIPANTS: 96 Aboriginal and 96 non-Aboriginal women with no known medical factors affecting fetal growth or gestation. SETTING: Four remote far north Queensland communities served by the Far North Regional Obstetric and Gynaecological Service (FROGS) and the antenatal clinic at Cairns Base Hospital. MAIN OUTCOME MEASURES: Neonatal birth weight, length, head circumference, abdominal circumference, mid-arm circumference and triceps skinfold thickness, and derived ponderal index (weight/length3), head:abdomen circumference ratio and head:mid-arm circumference ratio. RESULTS: The Aboriginal neonates were on average almost 450 g lighter than their non-Aboriginal counterparts. They were also slightly shorter, with smaller head, abdomen and mid-arm circumferences and lower mean ponderal indices and triceps skinfold thickness. The gestational characteristics of the two groups were not significantly different. Neonatal phenotype was significantly associated with maternal body mass index and maternal age in both groups. Alcohol use in pregnancy was associated with a significant reduction in Aboriginal neonatal size, while tobacco use was significantly associated with size reductions in non Aboriginal babies. CONCLUSIONS: Culturally appropriate antenatal care programs targeting the effects of poor nutrition and excessive alcohol use are needed if the excess incidence of low birthweight in Aboriginal people is to be reduced. PMID- 11419770 TI - Improving access to specialist care for remote Aboriginal communities: evaluation of a specialist outreach service. AB - OBJECTIVE: To identify barriers faced by Aboriginal people from remote communities in the Northern Territory (NT) when accessing hospital-based specialist medical services, and to evaluate the impact of the Specialist Outreach Service (SOS) on these barriers. DESIGN: Combined quantitative and qualitative study. SETTING: Remote Aboriginal communities in the "Top End" of the NT, 1993-1999 (spanning the introduction of the SOS in 1997). PARTICIPANTS: 25 remote health practitioners, patients and SOS specialists. MAIN OUTCOME MEASURES: Numbers of consultations with specialists; average cost per consultation; perceived barriers to accessing hospital-based outpatient care; and perceived impact of specialist outreach on these barriers. RESULTS: Perceived barriers included geographic remoteness, poor doctor-patient communication, poverty, cultural differences, and the structure of the health service. Between 1993 and 1999, there were 5,184 SOS and non-SOS outreach consultations in surgical specialties. Intensive outreach practice (as in gynaecology and ophthalmology) increased total consultations by up to 441% and significantly reduced the number of transfers to hospital outpatient clinics (P< 0.001). Average cost per consultation was $277 for SOS consultations, compared with $450 at Royal Darwin Hospital and $357 at the closest regional hospital. Outreach has reduced barriers relating to distance, communication and cultural differences, and potentially bolsters existing primary healthcare services. CONCLUSIONS: When compared with hospital-based outpatient services alone, outreach is a more accessible, appropriate and efficient method of providing specialist medical services to remote Aboriginal communities in the NT. PMID- 11419771 TI - Has the ban on smoking in New South Wales restaurants worked? A comparison of restaurants in Sydney and Melbourne. AB - OBJECTIVE: To evaluate compliance with a legislative ban on smoking inside restaurants by comparing smoking in Sydney restaurants (where it is legally banned) with smoking in Melbourne restaurants (not subject to a legal ban). DESIGN AND PARTICIPANTS: Unobtrusive observational study of restaurant patrons, and interviews with restaurant staff, carried out by 159 volunteers. SETTING: 78 Sydney restaurants with smoke-free indoor environments (as required by legislation) and 81 Melbourne restaurants not subject to legislation preventing smoking. The study took place from 20-31 October 2000. INTERVENTION: Legislation to ban smoking in indoor areas of restaurants was introduced in New South Wales in September 2000 (about six weeks before our study). OUTCOMES: Observed incidents of smoking inside restaurants; staff attitudes to the ban; customer satisfaction as indicated by comments to staff; staff perceptions of restaurant patronage. RESULTS: No restaurant patrons were seen smoking in 78 Sydney restaurants during 156 hours of observation of 2,646 diners, compared with 176 smokers among 3,014 Melbourne diners over 154 hours of observation. Thirty-one per cent (24/78) of Sydney restaurants had experienced smokers attempting to smoke indoors after the legislation was introduced; 6% (5/78) reported instances of smokers refusing to stop smoking when asked; 79% (62/78) of restaurants had received favourable comments from patrons about the smoke-free law; 81% (63/78) of restaurant staff interviewed either supported or strongly supported the law. Since introduction of the legislation, 76% of restaurants reported normal trade, 14% increased trade, and 9% reduced trade. CONCLUSIONS: Smoke-free restaurants do not require "smoking police" to enforce bans, present few ongoing difficulties for staff, attract many more favourable than unfavourable comments from patrons, and do not adversely affect trade. PMID- 11419772 TI - Ladders: accidents waiting to happen. AB - OBJECTIVES: To examine the nature, severity and outcomes of injuries sustained from ladder falls. DESIGN: Retrospective survey of medical records. PATIENTS AND SETTING: Patients who presented after a fall from a ladder to the emergency department of a tertiary hospital in Melbourne, Victoria, between January 1994 and December 1997. MAIN OUTCOME MEASURES: Demographic characteristics; height and mechanism of fall; injury site and Injury Severity Score (ISS); rate of hospital admissions. RESULTS: 163 patients presented after ladder falls. They were aged 2.5 to 86 years (mean age, 48 years); 83% were male, and 78% were injured in non occupational settings. Almost half the accidents (43%) were caused by ladder instability (ladder sliding from position or tilting sideways). Most patients had mild or moderate injuries (usually of the extremities), but 13% had an ISS > or = 16 (indicating severe trauma), usually with head, chest or spinal injuries; 42% were admitted to hospital. Multiple regression analyses showed that ISS increased signficantly with height of fall and age (P< 0.05 for both), although a substantial amount of variation was not attributable to these variables. Likelihood of hospital admission increased linearly with increasing ISS to an ISS of 8 and remained high thereafter. CONCLUSIONS: Ladder falls resulted in significant morbidity, with men undertaking non-occupational activities comprising most of those injured. New strategies to encourage safe ladder use are needed. PMID- 11419773 TI - Caffeine-induced cardiac arrhythmia: an unrecognised danger of healthfood products. AB - We describe a 25-year-old woman with pre-existing mitral valve prolapse who developed intractable ventricular fibrillation after consuming a "natural energy" guarana health drink containing a high concentration of caffeine. This case highlights the need for adequate labelling and regulation of such products. PMID- 11419774 TI - Assessing competence to refuse medical treatment. AB - Doctors have a duty to ensure that patients in their care who refuse medical treatment are competent to do so. Doctors who comply with an incompetent patient's refusal of treatment risk harm to the patient and are breaching their duty of care. On the other hand, doctors who treat a competent patient against the patient's will violate his or her autonomy and commit a battery. The test of competence is a legal one, with the courts being the final arbiters in cases of doubt. Assessment of competence involves a three-step process of determining whether the patient (a) can comprehend and retain relevant information, (b) believes the information, and (c) can weigh the information in the balance and arrive at a choice. Asking patients a few simple questions can help doctors assess their competence. PMID- 11419775 TI - Evidence-based medicine in intensive care. AB - The life-threatening nature of critical illness, requiring simultaneous, multiple interventions, makes it difficult, if not impossible, to study the effects of any one treatment. It is often not possible to conduct trials in critically ill patients, as they can not give informed consent. Some high quality, prospective studies have influenced clinical practice in intensive care, but others with lower grades of evidence have led to some controversy. In intensive care, clinical practice is still influenced by a combination of theory, experience and evidence. PMID- 11419776 TI - Evidence-based medicine in practice--surgery. AB - Initial reticence on the part of some surgeons, combined with a lack of, or unsuitability of, randomised controlled trial evidence, has slowed the uptake of the evidence-based revolution in surgery. An important positive step has been the establishment of the Australian Safety and Efficacy Register of New Interventional Procedures--Surgical (ASERNIP-S) to evaluate new surgical procedures in an evidence-based manner. To supplement information from systematic reviews, ASERNIP-S has been involved in national data collection, which is important for production and evaluation of clinical practice guidelines. Evidence based practice in surgery, as in other disciplines, must be adequately resourced if it is to be widely and successfully applied. PMID- 11419777 TI - Refusal of treatment. AB - Patients' thoughts, feelings and desires are communicated in a variety of ways, and require sympathetic, critical interpretation. Patients need clear, evidence based medical information so that they can make their own decisions about whether to consent to or refuse medical treatment. Treatment refusal may provide an opportunity to introduce patients to advance care planning. Unconscious motivations in doctors may obstruct good clinical decision-making. Although respect for the patient's responsibility to make healthcare decisions should be a condition of the clinical relationship, healthcare decision-making is a collaborative process. PMID- 11419778 TI - Epilepsy. AB - Epilepsy may be associated with major social and medical problems, and counselling of patient and family is essential for good management. The workup of a person with a seizure includes history, physical examination, and laboratory testing. An electroencephalogram is essential to help classify the seizure and epilepsy type. Neuroimaging (preferably by magnetic resonance imaging) helps to exclude a structural abnormality. Seizures can be controlled with a single drug (monotherapy) in 70% of patients. The incidence of drug side effects is increased if more than one drug is used. Pregnancy is associated with an increased risk of seizures in 30% of women with epilepsy. Frequent assessment throughout pregnancy is important. There is a slightly increased risk of congenital malformation associated with the antiepileptic drugs. Folic acid supplementation is advisable. PMID- 11419779 TI - Treatment alternatives for nocturnal leg cramps. PMID- 11419780 TI - GPs can not use randomisation for their own patients. PMID- 11419781 TI - The ethics of recruitment. PMID- 11419782 TI - Depression is complex and heterogeneous. PMID- 11419783 TI - Medications are more heavily promoted than psychotherapies. PMID- 11419784 TI - Doctors could know better. PMID- 11419785 TI - Low-dose aspirin as prophylactic therapy for muscle cramp. PMID- 11419786 TI - Vaccine storage. PMID- 11419787 TI - Getting to grips with heroin and other opioid use. PMID- 11419788 TI - Whose ethics? PMID- 11419789 TI - The rediscovery of methadone for cancer pain management. PMID- 11419790 TI - Outstanding Australians. PMID- 11419791 TI - The battle of the sexes after fertilization: behaviour of paternal and maternal chromosomes in the early mammalian embryo. AB - In the early diploid mammalian embryo, a father's chromosomes don't mix with the mother's until some time after fertilization. This topological genome separation is preserved up to the four-cell embryo stage and then gradually disappears. Unlike maternal DNA, sperm DNA arrives in an almost crystalline structure, heavily modified with methylcytosines (MeCs), which keep genes inactive. Compartmentalization of the nucleus according to parental origin may make it easier for the cellular machinery of the fertilized egg to revive the paternal chromosomes and to control paternal gene expression. Active zygotic demethylation of the paternal genome by a putative demethylase in the egg is a striking example for the battle of the sexes at the genomic level and beyond the single-gene level. It has important implications for genomic imprinting, and the establishment of genetic totipotency in fertilized eggs and in somatic cells during mammalian cloning. PMID- 11419792 TI - Asynchronous chromosome pairing in male meiosis of the rat (Rattus norvegicus). AB - Premeiotic and meiotic chromosome distribution was studied in rat testes suspensions by a triple-color fluorescent staining protocol which allows simultaneous visual inspection of two chromosomal targets highlighted by FISH together with immunostained SCP3 synaptonemal complex (SC) proteins which are marked by a third, composite color. Triple labeling with rat chromosome (RNO) 4q and 19p specific probes and SCP3 staining disclosed that homologs are separated in premeiotic and leptotene nuclei. Pairing of homologous chromosome regions commenced during early zygotene, with pairing of the small metacentric chromosomes 19 preceding that of the distal region of the long arm of RNO4. Our results show that homolog association occurs during zygotene of rat spermatogenesis, with small and large chromosomes showing a considerable asynchrony. Comparison with pairing progression in meiosis of other mammals suggests that asynchronous chromosome pairing reflects size differences within a complement. PMID- 11419793 TI - Chicken spindlin genes on W and Z chromosomes: transcriptional expression of both genes and dynamic behavior of spindlin in interphase and mitotic cells. AB - Contigs of genomic clones covering about 480 kb on the terminal region of the short arm of chicken W chromosome were obtained. By applying the exon trapping procedure on this whole region, a chicken homolog of spindlin gene, chSpin-W, was identified and subcloned. A counterpart gene, chSpin-Z, was found near the centromere on the long arm of Z chromosome. Although protein-coding regions of both genes are nearly identical, a part of the 3'-untranslated region is sufficiently different to distinguish the transcript of chSpin-W. Both chSpin-W and chSpin-Z are transcribed in early embryos. chSpin-Z is transcribed in various tissues of adult chickens, while chSpin-W is transcribed most prominently in ovarian granulosa and thecal cells. When female chicken embryonic fibroblasts were transfected with a cDNA construct for red fluorescent protein or green fluorescent protein-fused spindlin or FLAG-tagged spindlin, the expressed spindlin was co-localized with SUMO-1 in nuclear dots, ND10, in interphase cells, while the expressed spindlin was localized on entire chromosomes during mitosis. The localization of spindlin in ND10 reappeared after mitosis in daughter cell nuclei. A C-terminal region of spindlin was suggested to be required for the localization of spindlin to ND10. PMID- 11419794 TI - Karyotype relationships between distantly related marsupials from South America and Australia. AB - Reciprocal chromosome painting and G-banding were used to compare the karyotypes of three Australian marsupials (Sminthopsis crassicaudata, Macropus eugenii, Trichosurus vulpecula) and one South American marsupial (Monodelphis domestica). The results revealed only a limited number of rearrangements between these species and that the four karyotypes can be described as different combinations of fifteen conserved segments. Five chromosomes are totally conserved between M. domestica (pairs 1, 2, 5, 8 and the X) and the presumed 2n = 14 Australian ancestral karyotype, while M. domestica pairs 3 and 6 and 4 and 7 would have been involved in fusion/fission rearrangements. Chromosome comparisons are presented in a chromosome homology map. Although the species studied diverged 70 million years ago, the karyotype of Monodelphis domestica is highly conserved in relation to those of Australian marsupials. PMID- 11419795 TI - Transition between two forms of heterochromatin at plant subtelomeres. AB - The manner of packing of the terminal DNA loci into nucleosomes and higher order structures may strongly influence their functional interactions. Besides the structural flexibility of telomeric DNA sequences, conserved features of their chromatin including short nucleosome phasing (157 bp) and nucleosome sliding have been described previously. To gain a complementary knowledge of subtelomeres, we have analysed the chromatin structure of two subtelomeric tandem repeats from the plant Silene latifolia: X43.1 and 15Ssp. X43.1 shows two distinct nucleosome periodicities--157 and 188 bp. Preferred positions of its two nucleosomes have been mapped at both low and high resolution and the experimental results correspond to computer-predicted positions. 15Ssp is a newly-discovered sequence showing a telomere-associated position by PCR and a subtelomeric location by pulsed-field gel electrophoresis and fluorescence in situ hybridisation. Its 159 bp sequence unit shows a tandem arrangement and the presence of micrococcal nuclease-hypersensitive sites when either naked DNA or chromatin is digested. Use of a chemical nuclease results in a regular nucleosome ladder of 157 bp periodicity. Moreover, 15Ssp mononucleosomes show instability and absence of specific positioning, features typical for telomeric chromatin. PMID- 11419796 TI - Target sites for SINE integration in Brassica genomes display nuclear matrix binding activity. AB - Short interspersed nuclear elements (SINEs) are ubiquitous components of complex animal and plant genomes. SINEs are believed to be important players in eukaryotic genome evolution. Studies on SINE integration sites have revealed non random integration without strict nucleotide sequence requirements for the integration target, suggesting that the targeted DNA might assume specific secondary structures or protein associations. Here, we report that S1 SINE elements in the genomes of Brassica show an interesting preference for matrix attachment regions (MARs). Ten cloned genomic regions were tested for their ability to bind the nuclear matrix both before and after a SINE integration event. Eight of the genomic regions targeted by S1 display strong affinity for the nuclear matrix, while two show weaker binding. The SINE S1 did not display any matrix-binding capacity on its own in either non-methylated or methylated forms. In vivo, an integrated S1 is methylated while the surrounding genomic regions may remain undermethylated or undergo methylation. However, tested genomic regions containing methylated S1, with or without methylated flanking genomic sequences, were found to vary in their ability to bind the matrix in vitro. These results suggest a possible molecular basis for a preferential targeting of SINEs to MARs and a possible impact of the integration events upon gene and genome function. PMID- 11419797 TI - FISH-mapping of 31 type I loci (Texas markers) to river buffalo chromosomes. PMID- 11419798 TI - Negotiation behavior when cultures collide: the United States and Japan. AB - This study compared the negotiation behaviors of Japanese and U.S. managers in intra- and intercultural settings. Transcripts from an integrative bargaining task were coded and analyzed with logistic and linear regression. U.S. negotiators exchanged information directly and avoided influence when negotiating intra- and interculturally. Japanese negotiators exchanged information indirectly and used influence when negotiating intraculturally but adapted their behaviors when negotiating interculturally. Culturally normative negotiation behaviors partially account for the lower joint gains generated by intercultural, relative to intracultural, dyads. The behavioral data inform motivational and skill-based explanations for elusive joint gains when cultures clash. PMID- 11419799 TI - On the dimensionality of organizational justice: a construct validation of a measure. AB - This study explores the dimensionality of organizational justice and provides evidence of construct validity for a new justice measure. Items for this measure were generated by strictly following the seminal works in the justice literature. The measure was then validated in 2 separate studies. Study 1 occurred in a university setting, and Study 2 occurred in a field setting using employees in an automobile parts manufacturing company. Confirmatory factor analyses supported a 4-factor structure to the measure, with distributive, procedural, interpersonal, and informational justice as distinct dimensions. This solution fit the data significantly better than a 2- or 3-factor solution using larger interactional or procedural dimensions. Structural equation modeling also demonstrated predictive validity for the justice dimensions on important outcomes, including leader evaluation, rule compliance, commitment, and helping behavior. PMID- 11419800 TI - The impact of self-efficacy on stressor-strain relations: coping style as an explanatory mechanism. AB - The present study was designed to examine whether coping style influences the impact of self-efficacy on stressor-strain relations. It was hypothesized that high self-efficacy would weaken stressor-strain relations when accompanied by frequent use of active coping and infrequent use of avoidance coping. Data collected from 2,293 members of the U.S. Army revealed 3-way interactions among self-efficacy, role clarity, and active coping and among self-efficacy, work overload, and avoidance coping. As predicted, self-efficacy mitigated the effects of low role clarity on strain only when active coping was high. Also as expected, strain levels were lower for participants with high self-efficacy than for participants with lower self-efficacy when work overload was low but avoidance coping was high. Implications of these findings for occupational stress research are discussed. PMID- 11419801 TI - Incremental validation of situational judgment tests. AB - Using 3 different samples, the authors assessed the incremental validity of situational judgment inventories (SJIs), relative to job knowledge, cognitive ability, job experience, and conscientiousness, in the prediction of job performance. The SJI was a valid predictor in all 3 samples and incrementally so in 2 samples. Relative to the other predictors, SJI's partial correlation with performance, controlling for the other 4 predictors, was superior in most comparisons. Subgroup differences on the SJI also appear to be less than those for cognitive ability and job knowledge, but greater than differences in conscientiousness. The SJI should prove to be a valuable additional measure in the prediction of job performance, but several additional areas of research are suggested. PMID- 11419802 TI - Organization justice evaluations, job control, and occupational strain. AB - To explore how organizational justice evaluations affect the occupational stress process, the authors formulated and tested the following 2 hypotheses: (a) The effect of job control on strain is mediated by justice evaluations, and (b) justice evaluations moderate the effect of job control on occupational strain. The results of structural equation modeling, based on data collected from 688 employees, suggest that job control affects strain through justice evaluations. Thus, the results of this study supported the 1st hypothesis and provided evidence that perceptions of the organization are potential factors contributing to employee health. PMID- 11419803 TI - Justice at the millennium: a meta-analytic review of 25 years of organizational justice research. AB - The field of organizational justice continues to be marked by several important research questions, including the size of relationships among justice dimensions, the relative importance of different justice criteria, and the unique effects of justice dimensions on key outcomes. To address such questions, the authors conducted a meta-analytic review of 183 justice studies. The results suggest that although different justice dimensions are moderately to highly related, they contribute incremental variance explained in fairness perceptions. The results also illustrate the overall and unique relationships among distributive, procedural, interpersonal, and informational justice and several organizational outcomes (e.g., job satisfaction, organizational commitment, evaluation of authority, organizational citizenship behavior, withdrawal, performance). These findings are reviewed in terms of their implications for future research on organizational justice. PMID- 11419804 TI - A note on the stability of team performance. AB - Yearly winning percentages of 23 professional basketball teams over a 10-year period were used to evaluate the stability of team performance. The intercorrelation matrix produced by these data is characterized by strong, positive correlations in adjacent time periods. As the number of intervening time periods increased, however, the observed correlations systematically decreased and ultimately became negative. Significant negative correlations of earlier performance with later performance are almost never observed with typical time related performance data. Possible explanations and boundary conditions for these atypical results are discussed. PMID- 11419805 TI - Traditional tests and job simulations: minority and majority performance and test validities. AB - Performance of 565 minority and 414 majority job applicants on traditional paper and-pencil tests and on a telephone simulation that measured similar constructs was assessed. Models of the measurement characteristics of these 2 test batteries indicated larger subgroup mean differences on the traditional tests than on the simulation. Correlations between traditional tests were lower in the majority sample than in the minority sample, and the variance of the minority candidates' scores on the traditional tests was much larger than the variance of majority applicants' scores on the same measures. The validity of the simulation was lower than the validity of the traditional tests. This study replicates previous laboratory research that has indicated smaller subgroup differences on simulations than on paper-and-pencil tests and extends this research by providing evidence of the relative validity of these 2 types of measures. PMID- 11419806 TI - Affective personality and the content of emotional social support: coping in organizations. AB - This field study extended previous research by simultaneously examining the influence of affective personality on 4 dimensions of emotional social support and job burnout. Furthermore, the dimensions of emotional social support were examined as to their differential effects on the components of burnout. Results suggest that affective personality characteristics are associated with emotional social support as well as burnout dimensions. Results also indicate that some types of emotional social support appear to guard against burnout, whereas other types appear to contribute to the burnout experience. These findings suggest that types of emotional social support may have different personality antecedents and that distinct dimensions of social support have differential consequences in regard to burnout. Suggestions for future research are offered. PMID- 11419807 TI - Implications of empirical Bayes meta-analysis for test validation. AB - Empirical Bayes meta-analysis provides a useful framework for examining test validation. The fixed-effects case in which rho has a single value corresponds to the inference that the situational specificity hypothesis can be rejected in a validity generalization study. A Bayesian analysis of such a case provides a simple and powerful test of rho = 0; such a test has practical implications for significance testing in test validation. The random-effects case in which sigma2rho > 0 provides an explicit method with which to assess the relative importance of local validity studies and previous meta-analyses. Simulated data are used to illustrate both cases. Results of published meta-analyses are used to show that local validation becomes increasingly important as sigma2rho increases. The meaning of the term validity generalization is explored, and the problem of what can be inferred about test transportability in the random-effects case is described. PMID- 11419808 TI - Toward a theory of individual differences and leadership: understanding the motivation to lead. AB - A broad, integrative theoretical framework for understanding the relationship between individual differences and various leader behaviors is presented; it proposes a new individual-differences construct called the motivation to lead (MTL). A large-scale study using 3 samples in different occupational and cultural contexts shows 3 factors underlying MTL, namely, affective-identity, noncalculative, and social-normative MTL. A parsimonious model of antecedents to MTL is developed through hierarchical regression modeling and is cross-validated using confirmatory latent variable modeling. MTL is shown to provide incremental validity over other predictors such as general cognitive ability, values, personality, and attitudes in the prediction of 2 behavioral measures of leadership potential. Findings are discussed with reference to the theoretical framework proposed for understanding individual differences in leader behavior. PMID- 11419809 TI - The job demands-resources model of burnout. AB - The job demands-resources (JD-R) model proposes that working conditions can be categorized into 2 broad categories, job demands and job resources. that are differentially related to specific outcomes. A series of LISREL analyses using self-reports as well as observer ratings of the working conditions provided strong evidence for the JD-R model: Job demands are primarily related to the exhaustion component of burnout, whereas (lack of) job resources are primarily related to disengagement. Highly similar patterns were observed in each of 3 occupational groups: human services, industry, and transport (total N = 374). In addition, results confirmed the 2-factor structure (exhaustion and disengagement) of a new burnout instrument--the Oldenburg Burnout Inventory--and suggested that this structure is essentially invariant across occupational groups. PMID- 11419810 TI - When openness to experience and conscientiousness are related to creative behavior: an interactional approach. AB - This study adopted an interactional approach to understanding how 2 of the Five Factor traits, openness to experience and conscientiousness, are related to creative behavior in the workplace. Openness to experience is theorized to result in high levels of creative behavior and conscientiousness is theorized to result in low levels of creative behavior when the situation allows for the manifestation of the trait influences. More specifically, the authors hypothesized that openness to experience would result in high levels of creative behavior if feedback valence were positive and job holders were presented with a heuristic task that allowed them to be creative. The authors also hypothesized that conscientiousness would result in low levels of creative behavior if supervisors engaged in close monitoring and coworkers were unsupportive. The authors tested their hypotheses in a sample of office workers, and 5 out of the 6 hypotheses were supported. PMID- 11419811 TI - "What else could he have done?" Creating false answers in child witnesses by inviting speculation. AB - In 2 experiments (N = 111 children), a suggestive technique for interviewing child witnesses called "inviting speculation" was examined. Children were presented with atypical actions for common objects in a clown show. One week later, the children were asked to speculate (e.g., "What else could he have done with the knife?") in a between-subjects design on all or none of the items (Experiment 1) and in a within-subjects design on part of the items (Experiment 2), thereby getting highly probable speculations (e.g., "to cut"). After a 3-week delay, the experimenters found more highly probable but not more other false answers for the experimental items (Experiment 2). After a 5-6-month delay, the rate of (unspecified) false answers increased compared with the baseline (Experiments 1 and 2). The short-term effect is explained by a speculation-as misinformation assumption, whereas the long-term effect is explained by the use of a metastrategy. PMID- 11419812 TI - The two faces of conscientiousness: duty and achievement striving in escalation of commitment dilemmas. AB - The author proposes that 2 facets of conscientiousness, duty and achievement striving, affect decision makers in escalation of commitment dilemmas in opposing ways, thus masking the predictive ability of a broad measure of conscientiousness. It is proposed that duty is associated with an other-centered orientation and that achievement striving is associated with a self-centered orientation. Analyses of decisions from 360 respondents showed that duty was associated with a deescalation of commitment, achievement striving was associated with an escalation of commitment, and the broad measure of conscientiousness was unassociated with commitment. The author advocates the utility of understanding potential self-centered versus other-centered aspects of the criterion of interest when conducting personality-based research. PMID- 11419813 TI - Treatment of gastroesophageal reflux disease: to step or not to step. PMID- 11419814 TI - What have we learned from the biological therapies in ulcerative colitis? PMID- 11419815 TI - Dieulafoy lesions: a review of 6 years of experience at a tertiary referral center. AB - Dieulafoy lesions are uncommon sources of GI hemorrhage and predominantly occur in the proximal stomach. At one time a pathological diagnosis made postoperatively, Dieulafoy lesions are now routinely diagnosed and treated endoscopically. Their true incidence is unclear as quiescent Dieulafoy lesions are easily overlooked on endoscopy and bleeding lesions are occasionally misidentified. Over 6 yr (June 1993-November 1999), 40 Dieulafoy lesions were identified on upper endoscopy at our institution, of which seven were located in the duodenum and one in the right colon. Forty-seven percent of patients were hospitalized for other causes before onset of bleeding, and 17 of 40 were found to have other abnormal findings at endoscopy. In 90% of the cases, endoscopic treatment was successful. Seven patients died, but none as a result of hemorrhage. In 24 endoscopically-treated patients in whom follow-up data are available, Dieulafoy bleeding recurred in one patient. Dieulafoy lesions are rare and often difficult to diagnose, but must be considered in the evaluation of upper and lower GI tract hemorrhage, as they can usually be managed endoscopically. PMID- 11419816 TI - Experience with juvenile polyps in North American children: the need for pancolonoscopy. AB - We report a recent experience with juvenile polyps (JP) in a large cohort of North American children to determine if a pancolonoscopy (PC) is needed in all children with suspected polyps. We reviewed hospital charts of all patients with JP seen over a 9-yr period (January, 1990-October, 1998). A total of 331 JP were encountered during 195 procedures in 184 patients (64% males, 88% white, mean age 5.93 yr [range 0.42-15.5 yr], median age 4.84 yr). Painless rectal bleeding was the commonest symptom. PC was performed in 42% (82/195) of procedures, and 177 JP were encountered: 54% (97/177) were in the rectosigmoid colon, 14% (24/177) were in the descending colon, and 32% (56/177) were proximal to the splenic flexure (i.e., proximal polyps). Overall, proximal polyps were seen in 37% (31/82) of PC. Only proximal polyps were noted in 12% (10/82) of PC. Five patients were re endoscoped after an initial limited examination because of continuing symptoms from proximal polyps. All but one of the polyps had typical features of a JP on histological examination. Though most JP are located in the left colon, a PC should be the initial procedure because: 1) 37% of PC revealed proximal polyps, 2) 32% of polyps were located proximal to splenic flexure, 3) persistence of symptoms from missed proximal polyp(s) necessitates a repeat study with attendant risks, and 4) there is a possibility of malignant transformation in an unidentified JP. PMID- 11419817 TI - Postmarketing reports of QT prolongation and ventricular arrhythmia in association with cisapride and Food and Drug Administration regulatory actions. AB - OBJECTIVE: To describe the postmarketing safety data used in the risk assessment of cisapride and to summarize the regulatory actions of the Food and Drug Administration (FDA). METHODS: The FDA analyzed reports of patients who developed QT prolongation, torsades de pointes, and ventricular arrhythmia in association with the use of cisapride to assess probable etiology and risk factors. RESULTS: While cisapride was being marketed from 1993-1999, the FDA received reports of the following patients: 117 who developed QT prolongation; 107, torsades de pointes; 16, polymorphic ventricular tachycardia; 18, ventricular fibrillation; 27, ventricular tachycardia; 25, cardiac arrest; 16, serious (unspecified) arrhythmia; and 15, sudden death; for a total of 341 individual patients affected, following use of cisapride. Eighty (23%) of the 341 patients died. Deaths were directly or indirectly associated with an arrhythmic event. Factors that suggested an association with cisapride included a temporal relationship between use of cisapride and arrhythmia, the absence of identified risk factors and other explanations for arrhythmia in some patients, and cases of positive dechallenge and rechallenge. In most individuals, the arrhythmia occurred in the presence of risk factors (other drugs and/or medical conditions). CONCLUSIONS: Postmarketing reports and pharmacokinetic and electrophysiological data provided evidence that cisapride is associated with the occurrence of QT prolongation and torsades de pointes. The risk of fatal arrhythmia with cisapride was believed to outweigh the benefit for the approved indication, treatment of nocturnal heartburn due to gastroesophageal reflux disease, leading to the drug's discontinuation in the United States. PMID- 11419818 TI - Management of heartburn in a large, randomized, community-based study: comparison of four therapeutic strategies. AB - OBJECTIVE: Our objective was to compare four management strategies for heartburn: therapy with an H2-receptor antagonist (ranitidine), therapy with a proton pump inhibitor (lansoprazole), crossover from ranitidine to lansoprazole ("step-up" therapy), and crossover from lansoprazole to ranitidine ("step-down" therapy). METHODS: This was a controlled, double-blind, multicenter trial comprising 593 adults with heartburn, randomized to one of four groups for 20 wk. Subjects received either ranitidine 150 mg b.i.d. for 20 wk, or lansoprazole 30 mg once daily for 20 wk, or ranitidine 150 mg b.i.d. for 8 wk [corrected] followed by lansoprazole 30 mg once daily for 12 wk ("step-up"), or lansoprazole 30 mg once daily for 8 wk followed by ranitidine 150 mg b.i.d. for 12 wk ("step-down"). Outcome measures were based on self-reports in daily diaries of 24-h heartburn severity, measured by maximum daytime and nighttime severity, and percentage of 24-h heartburn-free days measured by absence of both daytime and nighttime heartburn. RESULTS: Median heartburn severity was significantly lower (p < 0.05) for lansoprazole (0.25) than the other groups (0.46 ranitidine, 0.44 "step-up," 0.35 "step-down"). The lansoprazole group had a significantly higher percentage of 24-h heartburn-free days (median 81.4%, p < 0.01) than other groups (66.6, 66.9, and 73.6%, respectively). In the "step-up" and "step-down" groups, heartburn was less severe, and percentages of 24-h heartburn-free days were higher during lansoprazole treatment regardless of treatment sequence. CONCLUSION: Proton pump inhibitor treatment provides more consistent heartburn relief than an H2-receptor antagonist, or "step-up" or "step-down" therapy. PMID- 11419819 TI - Hiatal hernia size is the dominant determinant of esophagitis presence and severity in gastroesophageal reflux disease. AB - OBJECTIVE: Although reflux esophagitis is a multifactorial disease, the relative importance of these pathogenetic factors has not been clearly established. In this study, regression analysis was used to model the major determinants of esophagitis in patients with symptomatic gastroesophageal reflux disease (GERD). METHODS: Sixty-six GERD patients and 16 asymptomatic controls were evaluated. All patients underwent upper endoscopy, esophageal manometry, and 24-h pH monitoring. Esophagrams were performed in 38 of the GERD patients and all controls. Stepwise regression was performed using esophagitis severity as the dependent variable. Logistic regression was performed grouping subjects as controls, nonerosive GERD, or erosive esophagitis. RESULTS: Hiatal hernia size, lower esophageal sphincter pressure, esophageal acid exposure, and number of reflux episodes >5 min significantly correlated with esophagitis severity. Stepwise regression identified hiatal hernia size (p = 0.0001) and lower esophageal sphincter pressure (p = 0.0024) as significant predictors of esophagitis. Logistic regression also identified hiatal hernia size (chi2 = 17.07, p < 0.0001) and lower esophageal sphincter pressure (chi2 = 5.97, p = 0.0146) as significant predictors of erosive esophagitis. CONCLUSION: Esophagitis severity is best predicted by hiatal hernia size and lower esophageal sphincter pressure. Of these, hiatal hernia size is the strongest predictor. PMID- 11419820 TI - Prospective evaluation of esophageal motor dysfunction in Down's syndrome. AB - OBJECTIVE: The aim of this study was to determine the prevalence and way of presentation of esophageal motor dysfunction in a nonselected population of subjects with Down's syndrome. METHODS: The study was conducted in 58 Down's syndrome patients and 38 healthy controls. A global symptom score and individual scores for dysphagia for liquids and solids, heartburn, vomiting/regurgitation, and chest pain were obtained. Esophageal function was evaluated initially by scintigraphy using liquid and semisolid bolus. Time-activity curves based on the mean condensed images were used to calculate residual activity at 100 s after swallowing. According to both scintigraphy and clinical evaluation results, participants underwent a radiological and manometric study. RESULTS: The most frequent symptoms in Down's syndrome patients were: dysphagia for liquids (n = 9), dysphagia for solids (n = 10), vomiting/regurgitation (n = 8), and chest pain (n = 2). Liquid and semisolid retention of the tracer was significantly higher in Down's syndrome patients than in controls (p < 0.05). In 15 participants with Down's syndrome, tracer retention was higher than the 95 percentile of controls' retention. No correlation was found between the global or individual symptom score and esophageal retention quantified by scintigraphy. Hypothyroidism was unrelated to esophageal symptoms or retention. Five of the 15 esophagograms performed were abnormal, showing barium retention and/or esophageal dilation. Manometry showed achalasia in two subjects, total body aperistalsis in one, and nonspecific esophageal motor disorder in two. CONCLUSION: Esophageal motor disorders, particularly achalasia, are frequent in individuals with Down's syndrome. Awareness of esophageal dysmotility in this population is important, even though symptoms are not evident, to avoid potential complications. PMID- 11419821 TI - Effect of intragastric volume and osmolality on mechanisms of gastroesophageal reflux in children with gastroesophageal reflux disease. AB - OBJECTIVE: Both transient lower esophageal sphincter (LES) relaxations (TLESRs) and periods of low/absent LES pressure (LESP) are the main mechanisms of gastroesophageal reflux. These events are believed to be triggered by stimuli from different areas of the upper GI tract. We aimed at investigating the relationship between LESP profile and gastric emptying and distension after meals of different composition in 30 children with gastroesophageal reflux disease (median age 7.0 yr, range 12 months-12 yr). METHODS: Recordings of LESP and intraesophageal pH for 1 h fasting and for 2 postprandial h were performed with a perfused sleeve catheter and flexible electrode, respectively; gastric emptying and distension of antral area were simultaneously recorded with real-time ultrasonography. Ten patients had a standard meal (group A), 10 had a high-volume meal (group B), and 10 had a high-volume and osmolality meal (group C). RESULTS: Postprandial esophageal acid exposure was significantly higher in patients of groups B and C than in patients of group A (p < 0.01); it was also more prolonged in patients of group C than in subjects of group B (p < 0.05). A higher postfeeding rate of reflux episodes caused by TLESRs was detected in patients of groups B and C as compared with patients of group A (p < 0.01). This increase did not statistically differ in patients of groups B and C. Patients of group C exhibited a higher postprandial rate of reflux episodes associated with low/absent tone of the LES as well as a more prolonged gastric emptying time and a higher postfeeding gastric distension as compared with patients of groups A and B (p < 0.01). Finally, a significant correlation was only found between the postprandial rate of reflux events resulting from low/absent LESP and the degree of antral distension in patients of group C (p < 0.01). CONCLUSION: Gastroesophageal reflux is worsened by increasing the volume and osmolality of meals through significant changes of LESP. Meals of high volume and meals with high volume and osmolality cause a comparable increase of reflux episodes as a result of TLESRs. However, meals with high volume and osmolality cause the higher degrees of esophageal acid exposure than meals with high volume resulting from a higher rate of reflux episodes associated with low/absent LESP. This finding correlates with a high postfeeding antral distension. PMID- 11419822 TI - Coexpression of vascular endothelial growth factor and p53 protein in squamous cell carcinoma of the esophagus. AB - OBJECTIVE: p53 plays a role in tumor angiogenesis, and vascular endothelial growth factor (VEGF) plays a key role in tumor angiogenesis. The aim of the present study was to clarify how expression of p53 protein participates in angiogenesis, and whether the coexpression of VEGF and p53 protein has a significance for angiogenesis and the clinicopathological features in esophageal squamous cell carcinoma (SCC). METHODS: Tissues samples were taken from 60 patients with esophageal SCC after surgery. The expression of VEGF and p53 protein in these SCC was examined immunohistochemically. Microvessel density (MVD) was determined by counting microvessels in tumor sections stained for Factor VIII-related antigen. Ki-67 labeling index (LI) was calculated, based on Ki-67 antigen immunostaining, as a proliferative marker. Apoptotic index (AI) was calculated, based on the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick end labeling, to evaluate apoptosis. RESULTS: VEGF expression was observed in 58.3%, and p53 protein expression was observed in 61.7% of the 60 patients. VEGF and p53 protein were significantly coexpressed in 26 (43.4%). Histological venous invasion (p < 0.01) and distant metastasis (p < 0.05) were significantly correlated with p53 protein expression. The two parameters were more frequently observed in the SCC with VEGF/p53 coexpression than in those without the coexpression. The MVD and Ki-67 LI were significantly higher (p < 0.01 and p < 0.001), and the AI was significantly lower (p < 0.001) in the SCC with p53 protein expression than in the SCC without it. The MVD and Ki-67 LI were higher, and the AI was lower in the SCC with VEGF/p53 coexpression than in those without the coexpression. The 5-yr survival rate in patients with the coexpression was poorer than in the other patients. CONCLUSION: These results suggest that mutant p53 expression is associated with angiogenesis and distant metastasis in esophageal SCC, and that the coexpression of p53 and VEGF may play an important role in angiogenesis, and have important clinical significance. PMID- 11419823 TI - Simplified 13C-urea breath test for detection of Helicobacter pylori infection. AB - OBJECTIVE: The U.S. standard 13C-urea breath test (13C-UBT) has proven to be extremely reliable but entails several complicated performance requirements and a test period of approximately 1 h. The aim of this study was to compare the standard 13C-UBT with a simplified version embodying modifications of test meal, duration of fasting, amount of 13C-urea, method of breath collection, and duration of test. METHODS: This was a randomized, three-way, crossover study of the standard U.S. 13C-UBT, which contains 125 mg of 13C-urea and a pudding test meal. The final breath sample is taken 30 min after urea ingestion. This test was compared with a formulation containing 75 mg of 13C-urea, a 2.5-g citric acid test meal (UBT-Lite), and a final breath sample taken by direct exhalation into tubes 15 min after urea ingestion. We also compared the effect of prior meals versus fasting on the test outcome with the UBT-Lite. RESULTS: A total of 259 subjects were enrolled in the trial, and 249 completed all three urea breath tests. There was excellent agreement between the three versions of the UBT with >98% of subjects having concordant results. Using predetermined criteria, there was substantial equivalence between the tests. Neither solid and/or liquid food up to 1 h before performing the UBT-Lite affected outcome. CONCLUSION: The UBT Lite formulation of the 13C-UBT proved to be an improved version of the U.S. standard 13C-UBT offering less expensive ingredients, shorter test duration, and a simplified breath test collection method, without sacrificing accuracy. PMID- 11419824 TI - Pathological disorders of the gastric mucosa surrounding carcinomas and primary lymphomas. AB - OBJECTIVE: Gastritis, intestinal metaplasia, atrophy, and dysplasia are disorders that frequently precede the full development of gastric adenocarcinoma. On the other hand, primary gastric lymphomas seem to arise from mucosa-associated lymphoid tissue. It is well accepted that these histological changes are caused by Helicobacter pylori infection. The objective of this study is to determine the frequency and characteristics of epithelial and lymphoid tissue disorders of the gastric mucosa surrounding primary carcinomas and lymphomas. METHODS: We studied 111 gastrectomies from patients harboring primary adenocarcinomas (30 intestinal and 30 diffuse type) and 51 gastric lymphomas. For comparative purposes, we analized 86 stomachs from patients who died of diseases other than gastric malignancies. Histopathological disorders of the gastric mucosa adjacent to primary neoplasms such as atrophy, intestinal metaplasia, and dysplasia were recorded. Lymphoid follicles were classified in two groups, with or without expansion. Expansion was characterized by increased size, irregular borders, enlarged marginal zone, and expanded germinal centers. Differences were statistically evaluated with chi2 and Fisher exact tests, odds ratio, and relative risk, with 95% CI. p values <0.05 were considered statistically significant. RESULTS: Most intestinal-type adenocarcinomas showed atrophy (76.6%) and intestinal metaplasia (86.6%) and less frequently, dysplasia (23.3%), in the surrounding gastric mucosa. Expansive lymphoid follicles were more frequent among lymphomas than in adenocarcinomas (56.8% vs 25%); however, a high percentage of lymphomas were also associated with atrophy (50.9%), intestinal metaplasia (62.7%), and rarely dysplasia (11.8%). On the contrary, diffuse-type adenocarcinoma displayed less frequently atrophy (33%), intestinal metaplasia (50%), and dysplasia (3%). Gastric mucosa from patients without any gastric neoplasia was almost normal (84%), whereas the remaining 16% showed, both or alone, atrophy and intestinal metaplasia. CONCLUSION: Histopathological disorders of the gastric mucosa are not specific for any neoplasm, but intestinal-type adenocarcinomas frequently showed atrophy, intestinal metaplasia, and not uncommonly, dysplasia of the surrounding non-neoplastic gastric mucosa. Diffuse type adenocarcinomas did not frequently show such lesions. Primary lymphomas displayed expansive lymphoid follicles and also a high percentage of atrophy and intestinal metaplasia of the surrounding gastric mucosa. The presence of intestinal metaplasia, atrophy, and lymphoid follicles with expansion in endoscopic biopsies could suggest a higher suceptibility for the development of gastric intestinal-type adenocarcinoma or gastric lymphoma. Patients harboring such histopathological changes must receive eradication therapy against H. pylori and probably closer follow-up. PMID- 11419825 TI - Role of Helicobacter pylori infection in gastroduodenal injury and gastric prostaglandin synthesis during long term/low dose aspirin therapy: a prospective placebo-controlled, double-blind randomized trial. AB - OBJECTIVES: Whether gastric infection with Helicobacter pylori increases the risk of gastric mucosal injury during long term/low dose aspirin therapy is unknown. We examined whether H. pylori infection enhances upper GI mucosal damage, assessed endoscopically, in volunteers given low dose aspirin. We studied 61 healthy men and women, 29 with and 32 without active H. pylori infection. METHODS: We treated volunteers for 45 days with a placebo or aspirin (either 81 mg every day or 325 mg every 3 days). Gastroduodenal mucosal damage was then assessed by endoscopy, as was gastric histology and ex vivo gastric mucosal prostaglandin E2 and F2alpha synthesis rates. RESULTS: Erosive disease from low dose aspirin (erosions and/or ulcers) occurred in 50% of H. pylori-infected volunteers and in 16% of their noninfected counterparts (p = 0.02). Aspirin caused a significantly higher average mucosal injury score in the gastric antrum in H. pylori-infected participants than in noninfected subjects (p = 0.03), and two H. pylori-infected subjects developed antral gastric ulcers. Subjects with H. pylori gastritis treated with the placebo had nearly 50% higher gastric mucosal prostaglandin (E2 plus F2alpha) synthesis rates than their noninfected counterparts (108 +/- 6 ng/g/min versus 75 +/- 6 ng/g/min, p < 0.001). Aspirin reduced mucosal prostaglandin synthesis to similar levels in infected and noninfected participants. CONCLUSIONS: Long term/low dose aspirin therapy led to more gastric mucosal damage when H. pylori gastritis was present than when it was absent, despite similar degrees of gastric mucosal prostaglandin depletion. PMID- 11419826 TI - Helicobacter pylori eradication attenuates oxidative stress in human gastric mucosa. AB - OBJECTIVE: Helicobacter pylori infection causes gastric diseases, but the responsible mechanisms are not completely understood. They can involve DNA and tissue damage induced by reactive oxygen and nitrogen species. Our aim is to investigate the effects of bacterial eradication on oxidative stress by measuring changes of relevant markers. METHODS: Antral biopsies were obtained from 34 patients with chronic atrophic gastritis and peptic ulcer disease before and after bacterial eradication. The expression of inducible nitric oxide synthase (iNOS) and levels of nitrotyrosine (NTYR) and 8-hydroxy-2'-deoxyguanosine were assessed immunohistochemically as markers of nitric oxide (NO) production and of damage to proteins and DNA, respectively. RESULTS: Before treatment, the percentages of patients with staining were: 56 for iNOS in inflammatory cells, 79 and 61 for NTYR and 8-hydroxy-2'-deoxyguanosine in foveolar cells, respectively, and 82 for 8-hydroxy-2'-deoxyguanosine in lymphoid follicles. NTYR staining was associated with the intensity of inflammation (p = 0.04) and gastritis activity (p = 0.07). The prevalence of 8-hydroxy-2'-deoxyguanosine tended to be associated with that of NTYR. After successful H. pylori eradication, the prevalence of iNOS and NTYR (in mild gastritis) staining decreased (p < 0.001 and p < 0.06, respectively). 8-Hydroxy-2'-deoxyguanosine staining disappeared in 24% of cases but appeared in 18% of previously negative cases despite eradication. CONCLUSION: Targets of oxidative stress associated with H. pylori infection are inflammatory and deep foveolar cells and lymphoid follicles. This is the first report of 8 hydroxy-2'-deoxyguanosine localization in gastric mucosa. Oxidative stress is reduced by bacterial eradication in the first stages of mild gastritis. Moderate severe gastritis may be a step that is reversible for iNOS, but partly irreversible for NTYR and 8-hydroxy-2'-deoxyguanosine. PMID- 11419827 TI - Pantoprazole therapy in the long-term management of severe acid peptic disease: clinical efficacy, safety, serum gastrin, gastric histology, and endocrine cell studies. AB - OBJECTIVE: Pantoprazole is the third proton pump inhibitor to become available. When this study was started, there were few data on its long-term use. Our aim was to investigate this aspect and, because powerful inhibitors of acid secretion can cause hypergastrinemia and, in experimental animals, enterochromaffin-like cell hyperplasia, we also monitored serum gastrin and endocrine cell histology. METHODS: One hundred fifty patients refractory to H2-receptor antagonists, running an aggressive course or with complications, were entered into a 5-yr treatment program. We performed serial endoscopy, checked for adverse events, and laboratory values. We also monitored serum gastrin, gastric endocrine cell histology, and antral and corpus gastritis. RESULTS: This report presents results from up to 3 yr of treatment. Cumulative healing on 40-80 mg of pantoprazole was 82% at 4 wk and 92% by 12 wk. Most patients became asymptomatic within 4 wk. Remission on maintenance treatment with 40 mg (n = 111) was 85% at 12 months and 78% at 24 months. Treatment was safe; only four patients had adverse events definitely related to pantoprazole. Elevations in gastrin were modest and there were no significant changes in gastric endocrine cells. The number of enterochromaffin-like cells tended to decrease. CONCLUSION: Pantoprazole is effective, safe, and does not seem to be associated with large increases in serum gastrin or alterations in gastric endocrine cells. PMID- 11419828 TI - Usefulness of carbohydrate-deficient transferrin and trypsin activity in the diagnosis of acute alcoholic pancreatitis. AB - OBJECTIVE: The aim of this study was to assess if carbohydrate-deficient transferrin (CDT) and trypsin activity differentiate acute alcoholic pancreatitis from nonalcohol-related pancreatitis, and as a secondary goal to evaluate its use in comparison to healthy controls. METHODS: Serum levels of CDT and trypsin activity were measured in frozen sera from 70 nonconsecutive patients with acute pancreatitis and in 16 healthy controls. RESULTS: Causes of pancreatitis were gallstones in 51%, chronic alcoholism in 23%, and other or unknown causes in 26% of the patients. Serum CDT was significantly higher in alcoholic pancreatitis than in the nonalcoholic disease (p < 0.0001) with a median (interquartile range) of 30.8 U/L (23.6-41.7 U/L) in chronic alcoholism, 16.7 U/L (13.05-21.1 U/L) in gallstones, 17.5 U/L (15.9-21.6 U/L) in unknown cause, 19.3 U/L (15.1-27.7 U/L) in other etiologies, and 16.1 U/L (12.1-18.8 U/L) in controls. At a cutoff over 22.5 U/L, CDT showed a sensitivity of 87.5% and a specificity of 85.2%. Serum levels of trypsin activity were significantly higher (p = 0.0007) in alcoholic pancreatitis, median 165 U/L (76-405 U/L) than in nonalcoholic pancreatitis, median 73 U/L (46.5-100.5 U/L). At a cutoff value over 152 U/L, the sensitivity of trypsin activity was 60% with a specificity of 100%. In the multivariate analysis, patient's age (< or = 44 yr), CDT (>22.5 U/L), and trypsin activity (>152 U/L) enabled correct prediction of acute alcoholic pancreatitis in 98% of the cases. CONCLUSION: Serum CDT and trypsin activity are of clinical utility in differentiating alcoholic from nonalcoholic acute pancreatitis. PMID- 11419829 TI - The utility of urgent colonoscopy in the evaluation of acute lower gastrointestinal tract bleeding: a 2-year experience from a single center. AB - OBJECTIVE: Urgent colonoscopy is often recommended to evaluate acute rectal bleeding. However, it may not identify a source because of blood in the lumen or inadequate preparation. Our aim was to determine the utility of urgent colonoscopy as the initial test for acute rectal bleeding. METHODS: This was a retrospective chart review of all patients discharged in 1997 and 1998 with an International Classification of Diseases, 9th Revision, code for hematochezia or rectal bleeding. RESULTS: We identified 514 charts but excluded 424 because of inaccurate coding. In the 90 with confirmed acute rectal bleeding, colonoscopy was the initial test in 39; age, sex, and race distributions were similar to those who did not have colonoscopy. A definite source of bleeding was seen at colonoscopy in only three patients, a probable source in 26, and no source in 10. Therapeutic intervention in four patients with a definite or probable source was successful in three. The commonest reasons for not performing urgent colonoscopy were bleeding from presumed hemorrhoids or bleeding that was clinically insignificant. Spontaneous resolution of bleeding and length of hospital stay were not affected by urgent colonoscopy. Five patients had surgery for unrelated reasons. In-hospital mortality was 2% and was unrelated to bleeding. CONCLUSION: Urgent colonoscopy as the initial investigation in acute lower GI tract bleeding probably does not alter the outcome in most cases. Identification of a definite bleeding source leading to successful therapeutic intervention is rare. Spontaneous resolution is frequent, length of hospital stay is similar, and clinical outcome is excellent regardless of whether or not urgent colonoscopy is performed. PMID- 11419830 TI - A patient education program is cost-effective for preventing failure of endoscopic procedures in a gastroenterology department. AB - OBJECTIVE: The growing use and complexity of endoscopy procedures in GI units has increased the need for good patient preparation. Earlier studies in this area have focused on the psychological benefits of patient education programs. The present study was directed at determining cost-effectiveness of a patient education program. METHODS: A prospective, randomized, controlled design was used. The patient population consisted of 142 patients aged 18-90 yr referred for an endoscopy procedure. Ninety-one (64%) participated in a targeted educational session conducted by a dedicated departmental nurse (group 1), 38 (27%) did not (group 2), and 13 (9%) received telephonic instruction (group 3). Before the endoscopy, all patients completed a questionnaire covering background data, endoscopy-related variables, anxiety level, and satisfaction. Patient cooperation and success/failure of the procedure were documented by the attending nurse. RESULTS: Male gender, previous endoscopy, and explanation from the referring physician were associated with a low level of anxiety (p < 0.05). There was a significant association between attendance in the education program and success of the endoscopy (p = 0.0009). Cancellations of procedures because of poor preparation occurred in 4.39% of group 1 in comparison with 26.31% and 15.38% of groups 2 and 3, respectively (p = 0.005). The overall cost of the procedure was reduced by 8.6%, 8.9%, and 5.5% for gastroscopy, colonoscopy, and sigmoidoscopy, respectively. All participants expressed satisfaction with the brochure. CONCLUSION: A pre-endoscopy patient education program apparently increase patient compliance, thereby decreasing both the need for repeated examinations and their attendant costs. PMID- 11419831 TI - A prospective, randomized, controlled trial of covered expandable metal stents in the palliation of malignant esophageal obstruction at the gastroesophageal junction. AB - OBJECTIVE: Palliation of malignant esophageal obstruction is an important clinical problem. Expandable metal stents are a major advance in therapy, but many stents become obstructed because of tumor ingrowth. The aim of this study was to compare a new, membrane-covered expandable metal stent to conventional prostheses in a randomized controlled trial. METHODS: Sixty-two patients with malignant inoperable esophageal obstruction at the gastroesophageal junction participated in the study. Patients were randomly assigned to covered or uncovered stents. The principal outcome measure was the need for reintervention because of recurrent dysphagia or migration. Secondary endpoints were relief of dysphagia measured by a dysphagia score (grade 0 = no dysphagia, grade 1 = able to eat solid food, grade 2 = semisolids only, grade 3 = liquids only, grade 4 = complete dysphagia) and the rate of complications and functional status. All patients were observed at monthly intervals until death or for 6 months. RESULTS: One week after stenting the dysphagia score improved significantly in both the uncovered (n = 32, 3 +/- 0.1 to 1 +/- 0.1 [means +/- SEMs], p < 0.001) and covered (n = 30, 3 +/- 0.1 to 1 +/- 0.2 [means +/- SEMs], p < 0.001) stents. Obstructing tumor ingrowth was significantly more likely in the uncovered stent group (9/30) than in the covered group (1/32) (p = 0.005). Significant stent migration occurred in 2/30 patients with uncovered stents, as compared with 4/32 patients in the covered group (p = 0.44). Reinterventions for tumor ingrowth were significantly greater in the uncovered stent group (27%), as compared with 0% in the covered group (p = 0.002). Life table analysis showed similar survival in both groups. CONCLUSION: Membrane-covered stents have significantly better palliation than conventional bare metal stents because of decreased rates of tumor ingrowth that necessitate endoscopic reintervention for dysphagia. PMID- 11419832 TI - Predictors of inadequate bowel preparation for colonoscopy. AB - OBJECTIVE: Inadequate preparation of the bowel for colonoscopy can result in both missed pathological lesions and cancelled procedures. We looked prospectively at the quality of colonic preparation and evaluated potential associations between specific patient characteristics and inadequate colonic preparation. METHODS: Data were gathered on consecutive patients presenting for colonoscopy who received either a polyethylene glycol lavage or oral sodium phosphate bowel preparation. Patient demographic and medical history information was gathered before scheduled colonoscopy. The endoscopist evaluated the preparation quality during the procedure. Complete data were gathered on 649 of 714 eligible patients (90.8%). Possible predictors of inadequate colonic preparation were analyzed using univariate statistics and multivariate logistic regression models. RESULTS: An inadequate colonic preparation was reported in 21.7% of observed colonoscopies. Only 18% of patients with an inadequate colonic preparation reported a failure to adequately follow preparation instructions. A later colonoscopy starting time, a reported failure to follow preparation instructions, inpatient status, a procedural indication of constipation, taking tricyclic antidepressants, male gender, and a history of cirrhosis, stroke or dementia were all independent predictors of an inadequate colon preparation (all p < 0.05). A procedural indication of previous polypectomy was a negative predictor of inadequate colonic preparation (p < 0.05). CONCLUSION: Several patient characteristics were significantly associated with colonic preparation quality independent of preparation type, compliance with preparation instructions, and procedure starting time. This information may help to identify patients at an increased risk for inadequate colonic preparation for whom alternative preparation protocols would be appropriate. PMID- 11419833 TI - Effectiveness of manual cleaning and disinfection of gastroendoscopes with 3% glutaraldehyde for decreasing risk of transmission of hepatitis C virus. AB - OBJECTIVES: The effectiveness of manual cleaning and disinfection of gastroendoscopes with 3% glutaraldehyde in decreasing the risk of transmission of hepatitis C virus (HCV) was examined. METHODS: Gastroendoscopes used for treatment of endoscopic esophageal variceal ligation in patients with HCV infection were manually cleaned and disinfected with 3% glutaraldehyde (n = 25), 2% glutaraldehyde (n = 17), or 0.1% benzethonium chloride (n = 25). Samples were obtained by pouring 20 ml of 0.9% NaCl solution into the biopsy-suction channel of the scope before and after cleaning and disinfection. HCV was detected with the polymerase chain reaction. RESULTS: Of the 25 scopes in 3% glutaraldehyde group, nine (36%) were positive for HCV before cleaning and disinfection, but all became negative after cleaning and disinfection; the difference was statistically significant (p < 0.01). In contrast, in 2% glutaraldehyde group and in the routine cleaning group, there were no significant differences in the number of positive samples between before and after cleaning and disinfection. CONCLUSION: Manual cleaning and disinfection of gastroscopes with 3% glutaraldehyde is useful for decreasing the risk of transmission of HCV among patients. PMID- 11419834 TI - An open-labeled, randomized study comparing systemic interferon-alpha-2A and prednisolone enemas in the treatment of left-sided ulcerative colitis. AB - OBJECTIVE: The aim of this study was to compare the treatment efficacies of subcutaneous interferon-alpha-2A (IFN-alpha-2A) injections versus prednisolone enemas in active left-sided ulcerative colitis in an open-labeled, randomized study. METHODS: Sixteen ulcerative colitis patients received IFN-alpha-2A subcutaneously (dosage: first wk, 9 MIU three times weekly [t.i.w.]; second wk, 6 MIU t.i.w.; wk 3-12, 3 MIU t.i.w.), and 16 received prednisolone enemas for 30 days (100 ml once daily, 0.25 mg of prednisolone/ml). The Powell-Tuck Index, Inflammatory Bowel Disease Questionnaire (IBDQ) score, and rectal histological activities were assessed before and after treatment. Thirteen patients in the IFN alpha-2A group and all 16 in the prednisolone enema group completed the treatment. RESULTS: IFN-alpha-2A treatment showed significant improvements in the Powell-Tuck Index (p = 0.0002), IBDQ score (p = 0.002), and rectal histological activity scores (p = 0.02). In the enema group, significant improvements were found in the Powell-Tuck Index (p = 0.0009), whereas no significant improvements were detected in the IBDQ scores (p = 0.055) or rectal histological scores (p = 0.052). There were no differences between scores of the two groups either before or after treatment. Only moderate side effects from the IFN-alpha-2A treatment were seen during the first 2-4 wk of treatment. CONCLUSION: IFN-alpha-2A treatment resulted in significant depression of the disease activity as reflected by the Powell-Tuck Index, IBDQ score, and histological disease activity scoring. The preliminary trial thus suggests that IFN-alpha-2A may be effective in the treatment of active left-sided ulcerative colitis. Larger, randomized trials are, however, warranted to confirm this finding, owing to possible type II errors in group comparisons. PMID- 11419835 TI - Concerns of patients with inflammatory bowel disease: results from a clinical population. AB - OBJECTIVE: The impact of chronic illness is influenced not just by physical symptoms but also by psychosocial factors. The aim of this study was to determine the concerns of inflammatory bowel disease (IBD) patients in a clinical sample, if concerns differ between patients from varied clinical and demographic variables, and if concerns influence well-being beyond the influence of physical symptoms. METHODS: Subjects (n = 259) completed a validated measure of concerns specific to IBD and provided demographic and disease-related information. RESULTS: The most intense concerns involved both physical (e.g., energy level) and psychosocial issues (e.g., achieving full potential). There were numerous differences in disease concerns based on ability to work but none based on disease duration. Factor analysis yielded three indices: body image and interpersonal concerns, general physical impact, and disease stigma. Age and education only affected certain concern indices in subgroups of patients. Greater concerns negatively influenced well-being beyond the influence of physical symptoms. CONCLUSION: Psychosocial factors, in addition to physical symptoms, play an important role on the impact of illness in patients with IBD. PMID- 11419836 TI - Cross-cultural variation in disease-related concerns among patients with inflammatory bowel disease. AB - OBJECTIVE: The aim of this work was to study cross-cultural variations in the impact of inflammatory bowel disease (IBD) on health-related quality of life by an international comparison of disease-related concerns. METHODS: Item and factor scores on the Rating Form of Inflammatory Bowel Disease Patient Concerns and overall mean concern levels were compared by analysis of variance among 2002 IBD patients in eight countries. RESULTS: The overall level of concern varied from 51 out of 100 in Portugal to 19 in Sweden, with intermediate scores for Italy (43), Canada (40), United States (39), France (39), Austria (33), and Israel (25). Having surgery, an ostomy, the uncertain nature of the disease, and medication side effects were each rated among the first five in importance in six countries. Other items varied considerably. For example, concern regarding pain and suffering was high in Israel and low in Portugal, whereas concern over developing cancer was low in Italy. Concern over financial issues and access to high-quality health care were inversely associated with measures of national economic prosperity. CONCLUSIONS: 1) Cross-cultural comparisons of patient concerns related to IBD are feasible using translated scales. 2) Reporting tendencies vary greatly; within Europe, patients from southern countries report greater overall concern. 3) The complications and the variable evolution of disease elicit general concern, but the importance of specific issues varies among countries. 4) The reasons for national differences may have social, cultural, and/or economic determinants with relevance to the patient-physician relationship, patient education, and therapeutic decision making. PMID- 11419837 TI - Prolonged multi-point recording of colonic manometry in the unprepared human colon: providing insight into potentially relevant pressure wave parameters. AB - OBJECTIVES: To determine the feasibility of and derive normative data for prolonged, 24-h, multipoint, closely spaced, water perfused manometry of the unprepared human colon. METHODS: In 14 healthy volunteers, 24-h recordings were made using a water perfused, balloon-tipped, 17 lumen catheter which was passed pernasally and positioned so that 16 recording sites spanned the colon at 7.5 cm intervals from cecum to rectum. The area under the pressure curve and propagating pressure wave parameters were quantified for the 16 regions. High amplitude propagating sequences were defined as were rectal motor complexes. RESULTS: Nasocolonic recording was well tolerated and achievable. Propagation sequences, including high amplitude propagating sequences, originated in the cecum (0.32 +/- 0.05/h) more frequently than in other regions and the extent of propagation correlated significantly with proximity of the site of sequence origin to the cecum (p < 0.001). Propagation velocity of propagating sequences was greater than high amplitude propagating sequences (p = 0.0002) and region-dependent, unlike high amplitude propagating sequences (p < 0.01). The frequency of propagating sequences did not increase after the meal, but frequency of high amplitude propagating sequences was increased significantly by the meal (p < 0.01). Rectal motor complexes were seen throughout the colon with no apparent periodicity. CONCLUSIONS: Prolonged, multipoint, perfusion manometry of the unprepared colon provides improved spatial resolution of colonic motor patterns and confirms the diurnal and regional variations in propagating pressure waves detected in the prepared colon. The study demonstrates differences between high amplitude propagating sequences and propagating sequence parameters that may have functional significance; and also, that the rectal motor complex is a ubiquitous pan colonic motor pattern. PMID- 11419838 TI - Attitudes toward smoking and smoking behaviors of patients with Crohn's disease. AB - OBJECTIVE: To examine the smoking behaviors of people with Crohn's disease. In active smokers, we measured their willingness to quit, their degree of nicotine dependence, and the proportion that made a quit attempt within 6 months to determine if they were refractory to smoking cessation in comparison to the general population. We also examined factors that were important in their decision to smoke. METHODS: We conducted a cross-sectional survey of out patients, supplemented by telephone interviews and a 6-month follow-up questionnaire of active smokers. Measures included disease activity, current smoking behaviors, intentions (stage of change), Fagerstrom Test for Nicotine Dependence, and factors related to their decision to smoke (decisional balance). RESULTS: The questionnaire was completed by 115 patients (78% response rate). Forty percent were active smokers. Of active smokers, 59% were considering quitting within the next 6 months, and of these, 15% were planning on quitting within the next 30 days. Those with moderate disease activity were more likely to be considering quitting than those with mild or severe activity. Nicotine dependence was rated as high in 33% and as moderate in 43%. Factors unrelated to Crohn's disease were more important in their decision to smoke than were Crohn's disease-related factors. After 6 months, 23% had made an attempt to quit and this attempt was strongly associated with their stated intentions at the baseline questionnaire. Two of three patients who had recently quit at baseline had resumed smoking. CONCLUSION: When compared to similar data for the general population, patients with Crohn's disease are no more refractory to smoking cessation. PMID- 11419839 TI - Comparison between color power Doppler ultrasound with echo-enhancer and spiral computed tomography in the evaluation of hepatocellular carcinoma vascularization before and after ablation procedures. AB - OBJECTIVE: Use of new echo enhancers capable of passing the lung filter has extended the clinical applications of color power Doppler flow imaging in many diseases and appears promising in the study of neoplasm vascularization. Levovist (Shering, Berlin Germany) is an ultrasound contrast agent containing galactose microbubbles suspended in palmitic oil. The sensitivity of Levovist-enhanced color power Doppler was compared to that of standard color power Doppler and contrast-enhanced spiral computed tomography (CT) in the detection of vascular signals in hepatocellular carcinoma. METHODS: We examined 29 hepatocellular carcinoma nodules in cirrhotic livers that had appeared avascular on unenhanced color power Doppler. Color power Doppler studies were repeated with and without Levovist enhancement before (15 examinations) and/or after (23 examinations) percutaneous ablation procedures. Findings (vascularized vs nonvascularized) were compared to those obtained with contrast-enhanced spiral computed tomography (gold standard) performed no more than 24 h after each of the 38 Doppler examinations. RESULTS: In pretreatment studies, Levovist-enhanced power Doppler correctly revealed vascularization in 12 of 15 lesions that had appeared avascular without echo enhancement and confirmed the avascularity of one other nodule; the remaining two, which appeared avascular on contrast-enhanced Doppler, displayed vascularization on the spiral CT examination. Levovist-enhanced power Doppler was fully concordant with spiral CT findings in all of the posttreatment examinations. CONCLUSION: Considering the absence of false positive results in this study, echo-enhanced color power Doppler can be considered reliable in diagnosing incomplete necrosis of hepatocellular carcinomas after percutaenous ablation. Spiral computed tomography can thus be reserved for those cases in which the enhanced power Doppler examination reveals no evidence of vascularity. PMID- 11419840 TI - Physical activity, biliary lipids, and gallstones in obese subjects. AB - OBJECTIVE: Gallstone disease is a major source of morbidity in the US. Reduced physical activity has been shown to be a risk factor for gallstone formation in recent studies; however, the mechanisms to explain how physical activity may protect against gallstone formation have not been well elucidated. We investigated the relationships between physical activity, biliary lipids, and gallstone disease. METHODS: Three types of habitual physical activity (work, sport, and leisure time), biliary lipids, and serum lipids were estimated or measured in 53 obese subjects undergoing gastric bypass surgery. These physical activities were defined as activity at work, sport activity during leisure time, and activity during leisure time excluding sports, respectively. RESULTS: We found that sport activity but not work and leisure time activities was inversely associated with gallstone disease. Lower levels of biliary bile salts and percent biliary bile salts (expressed in percentage of total biliary lipids) and higher levels of percent cholesterol were also found to be associated with gallstone disease. A lower level of sport activity appeared to be associated with higher levels of biliary cholesterol, percent biliary cholesterol, and serum triglycerides. CONCLUSION: Our findings demonstrate that low levels of physical activity are associated with gallstone formation. Our study also suggests that a possible mechanism for the protective effect of physical activity on gallstone formation is the lowering of biliary cholesterol levels, thus preventing cholesterol from precipitating in the bile. In addition, our data suggest that sport activity is a more effective form of physical activity than working and leisure activities in the prevention of gallstone disease. PMID- 11419841 TI - Prognostic value of hepatocyte proliferative activity after transjugular intrahepatic portosystemic shunt. AB - OBJECTIVES: Previous data indicated that the proliferating cell nuclear antigen labeling index (PCNA-LI) reflects the liver functional reserve in human liver cirrhosis. The aim of the study was to evaluate the hepatocyte proliferative activity as a marker for the outcome of patients after transjugular intrahepatic portosystemic shunt (TIPS). METHODS: Twenty-eight consecutive patients were electively treated with TIPS for recurrent variceal bleeding (n = 14), refractory ascites (n = 12), or hydrothorax (n = 2). PCNA immunostaining was analyzed on methanol-fixed, paraffin-embedded liver biopsies. RESULTS: After TIPS, six patients died within the first 3 months, eight other patients died later, two were transplanted, and 12 were alive at the time of analysis. Early death occurred in patients with refractory ascites (5/12) and/or in Child C patients (3/6). Among the evaluated variables, there was a statistical trend for the PCNA LI to be lower in patients who died early after TIPS than in those having long term survival (1.55% vs 2.65%, p = 0.07). After TIPS insertion, the probability of remaining alive during the first 6 months of follow-up was significantly higher in patients with a preprocedural PCNA-LI > 2.9%. CONCLUSIONS: The PCNA-LI measured on liver biopsy before the TIPS procedure might be a pre-TIPS marker to discriminate those patients for whom TIPS is likely to be beneficial. PMID- 11419842 TI - Hepatic familial amyloidosis caused by a new mutation in the apolipoprotein AI gene: clinical and pathological features. AB - OBJECTIVE: Recently, we reported a nondescribed deletion/insertion mutation in the apolipoprotein AI gene as the cause of hereditary amyloidosis with hepatic presentation. We describe the clinical and pathological features of this type of amyloidosis in one affected family. METHODS: Demographic, clinical, and biochemical data were obtained from 33 members of the family in whom the apolipoprotein AI gene was studied. Diagnosis was based on the detection of the apolipoprotein AI gene mutation, scintigraphy using radioionated serum amyloid P component, and histological and immunohistochemical studies. RESULTS: Eight members with the mutation had hepatic involvement. Six patients were practically asymptomatic, presented with an elevation of alkaline phosphatase and gamma glutamyl transpeptidase, and remained stable during follow-up (7.6 +/- 4.9 yr). One patient had jaundice, developed ascites and encephalopathy, and died of hepatorenal syndrome a few months after diagnosis. Jaundice and portal hypertension appeared in the remaining patient, who died 4 yr later. CONCLUSION: This form of familial amyloidosis is characterized by elevation in serum alkaline phosphatase and gamma-glutamyl transpeptidase secondary to amyloid deposits in the portal tracts. Patients remain stable and asymptomatic for many years, but portal hypertension and liver failure can develop later in life and lead to death. Thus, patients should be observed regularly and liver transplantation should be indicated when progression is detected. PMID- 11419843 TI - Usefulness of positron emission tomography with fluorine-18-fluorodeoxyglucose for predicting outcome in patients with hepatocellular carcinoma. AB - OBJECTIVES: The present study was designed to assess the usefulness of positron emission tomography with fluorine-18-fluorodeoxyglucose (FDG-PET) for predicting outcome in patients with hepatocellular carcinoma. METHODS: FDG-PET was performed in 48 patients with hepatocellular carcinoma. For quantitative evaluation, a region of interest (ROI) was placed over the area of maximum activity within the lesion. A background ROI was then placed over the nontumor region of the liver. The average activity within each ROI was subsequently corrected for radioactive decay, and the standardized uptake value (SUV) was calculated by dividing the tissue activity by the injected dose of radioactivity per unit body weight. SUV ratio was expressed as the tumor-to-nontumor ratio of the SUV. RESULTS: The tumor volume doubling time, as index of the growth rate of hepatocellular carcinoma, correlated significantly with SUV ratio but did not correlate with SUV. On the basis of the SUV ratio, the patients were divided into two groups of similar size: group A, SUV ratio of < or = 1.5; and group B, SUV ratio > 1.5. The cumulative survival rate was significantly lower in group B than in group A. On the basis of the SUV, the patients were divided into two groups of roughly equal size: group C, < or = SUV 2.6; and group D, > SUV 2.6. The cumulative survival rate was similar in these groups. On regression analysis with the Cox proportional hazards model, the SUV ratio and tumor number were significantly related to survival. CONCLUSIONS: These results suggest that FDG-PET is useful not only for the evaluation of the malignancy of hepatocellular carcinoma but also for the prediction of outcome in patients with hepatocellular carcinoma. PMID- 11419844 TI - Seasonal variations in variceal bleeding mortality and hospitalization in France. AB - OBJECTIVES: The circadian variation in portal blood pressure and in the diurnal incidence of variceal bleeding is well known, but the seasonal variation in variceal bleeding is still controversial. This report analyzes the seasonal variations in mortality and hospitalizations due to variceal bleeding in the French population. METHODS: All the deaths due to variceal bleeding that occurred from 1987 to 1996 (N = 13,514) and all adults discharged from French public hospitals for variceal bleeding from 1995 to 1997 (N = 17,026) were examined retrospectively. Cumulated monthly averages were expressed as the percentage above or below the average monthly value during the entire study period. RESULTS: Deaths due to variceal bleeding in France occurred with a clear annual periodicity and peaked in winter (December/January), both in the overall population and in subgroups defined by age and sex, except for women. The distribution of cumulative monthly deaths differed by 24%, with a peak (14% above average) in December and a trough (10% below average) in July (Roger's test: p < 0.001). Hospitalizations for variceal bleeding in French public hospitals followed a similar seasonal pattern (p < 0.001) with a winter-spring predominance (4% to 7% from December through April), except in patients aged 15-49 yr. There was a short sharp peak of mortality in early winter in French public hospitals. The seasonality of hospitalization and death increased markedly with age. CONCLUSIONS: A better understanding of these age- and sex-specific seasonal patterns would allow to improve pharmacological protection measures, disease management, and educational strategies. PMID- 11419845 TI - Quantitative analysis of hTERT mRNA expression in colorectal cancer. AB - OBJECTIVES: Telomerase is highly activated in a variety of malignant neoplasms including colon cancer. Among the major components of telomerase, human telomerase reverse transcriptase (hTERT) is thought to regulate telomerase activity. To assess the importance of telomerase for the diagnosis of colorectal cancer, we measured the expression of hTERT mRNA and telomerase activity in a large series of 140 colorectal cancers, 140 adjacent normal tissues, and 20 adenomas. METHODS: The expression level of hTERT was measured quantitatively by competitive reverse transcriptase-polymerase chain reaction (RT-PCR), and telomerase activity was examined by telomeric repeat amplification protocol (TRAP) assay in the same samples. RESULTS: The median expression level of hTERT mRNA in carcinomas was significantly higher than that in either adenomas or normal tissues. The median level of hTERT in adenomas was significantly higher than that in normal tissues. Telomerase activities in carcinomas were significantly higher than those in either adenomas or normal tissues. Telomerase activities in adenomas were also significantly higher than those in normal tissues. Furthermore, the relative expression levels of hTERT mRNA in adenomas and carcinomas were significantly correlated with the relative telomerase activities; the Spearman rank correlation was 0.53 (p = 0.021) and 0.18 (p = 0.031), respectively. CONCLUSIONS: Our data suggest that determination of hTERT mRNA by competitive RT-PCR is superior in quantitative accuracy and sensitivity and would support the importance of telomerase activity for the diagnosis of colorectal cancer. PMID- 11419846 TI - The effect of attending a flexible sigmoidoscopic screening program on the prevalence of colorectal adenomas at 13-year follow-up. AB - OBJECTIVES: Understanding the epidemiology of colorectal adenomas is a prerequisite for designing follow-up programs after polypectomy. The aim of the study was to investigate the effect of polypectomy on the long-term prevalence of adenomas. METHODS: In 1983, a total of 799 men and women aged 50-59 yr were drawn from the general population register. Of these, 400 comprised a screening group and 399 a matched control group. The screenees were invited to undergo a once only flexible sigmoidoscopy. Persons with polyps had a baseline colonoscopy with follow-ups in 1985 and 1989. In 1996, both the screenees and the controls were invited to a colonoscopic examination. RESULTS: In 1996, a total of 451 (71%) individuals attended. Adenomas were found in 78 (37%) individuals in the screening group and 103 (43%) in the control group, relative risk (95% confidence interval): 0.9 (0.7-1.1), p = 0.3, and high-risk adenomas (severe dysplasia, adenomas > or = 10 mm, villous components) were found in 16 (8%) and 32 (13%), respectively; relative risk (95% confidence interval): 0.6 (0.3-1.0), p = 0.07. CONCLUSIONS: There was no significant difference in adenoma prevalence between the group after the screening program and the controls after the usual care. There was a trend toward more high-risk adenomas in the control group. This suggests a very limited effect of one-time screening sigmoidoscopy with surveillance colonoscopy on the prevalence of adenomas, but a preventive effect on the development of high-risk adenomas consistent with the reported effect on cancer prevention. PMID- 11419847 TI - Fecal occult blood testing in a noncompliant inner city minority population: increased compliance and adherence to screening procedures without loss of test sensitivity using stool obtained at the time of in-office rectal examination. AB - OBJECTIVES: Fecal occult blood screening is cost-effective, is easily administered to large groups of patients, and reduces mortality associated with colorectal cancer. Within our predominant African American and Latino inner city clinic populations, compliance with common screening procedures is suboptimal. A procedure with increased compliance is needed to adequately screen this population at high risk for colorectal cancer. The objective of this study was to compare the results of the 3-day at-home hemoccult test for occult blood to those of a hemoccult test performed from stool obtained at rectal examination in the office. METHODS: A total of 350 consecutive patients referred to the GI clinic of University Hospital or Jersey City Medical Center for colorectal cancer screening had both the 3-day at-home hemoccult test and an in-office hemoccult examination performed, followed by either sigmoidoscopy (for negative results) or by colonoscopy (for positive results). RESULTS: Patients were noncompliant with dietary restrictions, 3-day card return, follow-up appointments, and endoscopy with conventional screening methods. Decisions based on the in-office examination with direct scheduling of endoscopy significantly improved compliance with follow up. There was no statistical difference between the two detection methods, suggesting that the in-office examination was the more effective screening test. CONCLUSIONS: Endoscopy based on an in-office hemoccult examination is an acceptable alternative to using the 3-day at-home stool collection to govern endoscopic choices. In a noncompliant inner city population, use of the in-office examination increased compliance with follow up, potentially allowing more patients exposure to screening. PMID- 11419848 TI - Is colorectal cancer screening necessary in the preoperative assessment of inguinal herniorrhaphy? A case-control study. AB - OBJECTIVES: The question of whether patients presenting for inguinal hernia repair require pre-operative assessment for colon cancer has remained unanswered. A case-control study is necessary to assess whether the prevalence of premalignant or malignant colonic lesions is higher in patients presenting with inguinal hernia compared to the general population. METHODS: Between 1990-2000, 614 inguinal herniorrhaphies were performed at the Veterans Affairs Palo Alto Health Care System (VAPAHCS). We retrospectively studied the 149 (24%) patients from this group with no prior history of colonic polyps, malignancy, or gastrointestinal bleeding who had flexible sigmoidoscopy or colonoscopy performed during the peri-operative period. Comparison was made to 149 controls undergoing colonoscopy or sigmoidoscopy during the same time period for colon cancer (CRC) screening. RESULTS: The mean (+/-SEM) patient age was 67 +/- 0.7 (range 31-92) yr in the hernia patients and 66 +/- 0.8 (range 46-93) in the control group (p = 0.7). Eighty-two of the inguinal hernia patients had screening procedures performed preoperatively with a mean time (+/-SEM) of 1.4 +/- 0.14 yr, while endoscopy was performed in the post-operative period for the remaining 67 patients (average time 2.7 +/- 0.2 yr, p < 0.001). More patients underwent colonoscopy in the control group compared to the hernia cohort (p = 0.004). Seven (5%) patients in the hernia group were found to have colorectal cancer compared to six (4%) in the control group (p = 0.8). CONCLUSIONS: This study does not support previously published findings that patients with inguinal hernias are more likely to have premalignant colonic lesions. Patients with inguinal hernias should undergo screening for colon cancer at the same rate as the general population. PMID- 11419849 TI - Proctoscope. PMID- 11419850 TI - Pancreatitis caused by common bile duct stones in a 3-year-old boy with prior surgery for a choledochal cyst. AB - Pancreatitis in children is uncommon. Compared to adults, pancreatitis in children is usually related to trauma, anatomic anomalies, infections, hereditary, and systemic disease, but not gallstones or alcohol. Most cases do not require endoscopic intervention. We report an unusual case of recurrent pancreatitis in a child related to common bile duct stones requiring endoscopic treatments after surgical treatment for choledochal cyst. PMID- 11419851 TI - Synchronous gastric adenocarcinoma and mucosa-associated lymphoid tissue lymphoma in association with Helicobacter pylori infection: comparing reported cases between the East and West. AB - The association of Helicobacter pylori infection with synchronous gastric adenocarcinoma and mucosa-associated lymphoid tissue (MALT) lymphoma is rare. Three Chinese patients (M:F = 1:2) who were 71, 58, and 75 yr of age were diagnosed to have gastric adenocarcinoma (2 patients) and gastric lymphoma (1 patient) on endoscopic biopsies. Distal gastrectomy was performed in all of them. Histological study of the three resected specimens revealed synchronous gastric adenocarcinoma and MALT lymphoma. H. pylori infection was found in two patients. A literature search revealed another 29 patients with synchronous tumors in whom H. pylori status was examined. Overall, H. pylori infection was found in 78% of 32 patients. The majority of lymphoma was low grade (75%) and was larger than carcinoma (81%). The majority of carcinoma (65.6%) was early. This suggested lymphoma might develop before carcinoma or the presence of MALT lymphoma might increase the risk of developing carcinoma. PMID- 11419852 TI - Simultaneous occurrence of inflammatory bowel disease and thyroid disease. AB - We report a case of simultaneous occurrence of inflammatory bowel disease (IBD) and Graves' disease. A review of reported cases of simultaneous onset of ulcerative colitis (UC) and autoimmune hyperthyroidism is presented. A discussion of the prevalence of thyroid disease in patients with UC and possible common autoimmune etiology is entertained. The concurrent presentation has implications for the diagnosis and treatment of both diseases. At the time of suspected initial presentation or exacerbation or preexisting IBD, we emphasize the need to consider both IBD and thyroid disease in the differential diagnosis for optimal patient management. PMID- 11419853 TI - Linear IgA bullous dermatosis responsive to a gluten-free diet. AB - Dermatitis herpetiformis is associated with a gluten-sensitive enteropathy in >85% of cases. Both the skin lesions and the enteropathy respond to gluten restriction. Linear IgA bullous dermatosis has a much lower prevalence of histological small bowel abnormalities, and lesions are not known to respond to gluten restriction. We report a patient with linear IgA bullous dermatosis and gluten-sensitive enteropathy. This report addresses the issue of whether linear IgA bullous dermatosis can be associated with gluten-sensitive enteropathy. We evaluated the response to gluten restriction and normal diet by following the status of the patient's jejunal biopsies and skin lesions. The patient responded to gluten restriction, as shown by resolution of jejunal abnormalities and skin lesions and subsequently by recurrence of jejunal abnormalities and skin lesions with reinstitution of a gluten-containing diet. This report demonstrates that linear IgA bullous dermatosis can respond to gluten restriction if an underlying gluten-sensitive enteropathy is present. PMID- 11419854 TI - Hope or hype--cytokine therapy in ERCP-induced pancreatitis. PMID- 11419855 TI - Surgery versus medical therapy for gastroesophageal reflux disease. PMID- 11419856 TI - Sphincter of Oddi manometry: is timing everything? PMID- 11419857 TI - Stool test for Helicobacter pylori. PMID- 11419858 TI - Re: Lanza et al.--Endoscopic comparison of alendronate and risedronate. PMID- 11419859 TI - Rofecoxib-induced renal dysfunction in a patient with compensated cirrhosis and heart failure. PMID- 11419860 TI - Age-related indications and complications after diagnostic laparoscopy. PMID- 11419861 TI - Reversible flaccid paraplegia after orthotopic liver transplantation. PMID- 11419862 TI - Detection of Helicobacter pylori with stool antigen test in children with gastroesophageal reflux disease. PMID- 11419863 TI - "Giant" colon lipoma: what kind of findings are necessary for the indication of endoscopic resection? PMID- 11419864 TI - Prevalence of irritable bowel syndrome and its relationship with Helicobacter pylori infection in a Japanese population. PMID- 11419865 TI - The portal flow velocity test is predictive of effective prevention of variceal bleeding. PMID- 11419866 TI - Report of a case showing a recovery from liver cirrhosis to chronic hepatitis, type C, after glycyrrhizin injection for 2 years and a sustained response by the following interferon therapy. PMID- 11419867 TI - Treatment of chronic hepatitis C in hereditary spherocytosis. PMID- 11419868 TI - Detection of esophagitis by technetium 99m tetrofosmin chest SPECT. PMID- 11419869 TI - The relevance of systemic complications and the different outcomes of subgroups after percutaneous endoscopic gastrostomy (PEG). PMID- 11419870 TI - A sudden coughing up of foul-smelling sputum: a first sign of a pancreaticobronchial fistula, a severe pulmonary complication in acute pancreatitis. PMID- 11419871 TI - A successful switch from prednisone to budesonide for neuropsychiatric adverse effects in a patient with ileal Crohn's disease. PMID- 11419872 TI - Closure of an enterocutaneous fistula by high dose intravenous immune globuline therapy in a patient with X-linked agammaglobulinemia. PMID- 11419873 TI - TEM-simulation of amorphous carbon films: influence of supercell packaging. AB - Recent developments in thin film technology allow to prepare deliberately amorphous carbon films with structures widely varying between graphite-like (sp2) and diamond-like (sp3) atomic bonds. This leads to amorphous structures with correspondingly varying densities. By periodically changing deposition conditions, nanometer multilayers may be prepared consisting of carbon layers of different density. Simulation of the electron microscopic imaging allows to differentiate between such real structural details (on the nanometer scale) and artefacts induced by the imaging procedure. But it must be assured that the modeled structure reflects the real one with sufficient accuracy. Thorough comparison of different simulation strategies shows that for the adequate simulation of TEM imaging of amorphous materials, the thickness of the layer with independently distributed atoms has to exceed a certain limit. Then, the statistical scattering of the randomly distributed atoms will be averaged. Otherwise, if the model of the transmission electron microscopy sample is constructed as iteration of thin identical supercells, the superposition of scattering waves with constant phase differences results in enhanced local fluctuations burying the multilayer structure. For thicker packages of supercells with independent random distributions, the effect of statistical atomic arrangements is more and more leveled off. Hence, nanometer structures based on regions with different density will be visible more distinctively in the random background. For carbon, this critical thickness amounts to about 4 nm. This is of special importance for the visualization of nanoscaled heterogeneities like multilayers or nanotube-like inclusions in amorphous matrices. PMID- 11419874 TI - Investigations of the interference of surface plasmons on rough silver surface by scanning plasmon near-field microscope. AB - A scanning plasmon near-field microscope with gold and silver tips, operating in tapping mode of atomic force microscope is used to measure the distribution of the near-field intensity of surface plasmons on rough silver surfaces. Using the fast Fourier transformation of near-field images, it is shown that the distribution of the near-field intensity on the surface is the result of the interference between scattering plasmons and the initial plasmon beam. Multiple scattering effects such as backscattering enhancement of surface plasmons are also observed. It is shown that a nonuniformity in the registration of the scattered light leads to some artifacts in near-field images. Several registration modes of the light signal are considered and it is shown that recording the light signal at the second harmonic of the tapping frequency one can pick out the signal associated with an electromagnetic (em) resonance in a tip-surface (sphere-plane) structure. Possible implementations of this em resonance for studies of local permittivities and local nonlinear susceptibilities of intermediate media between the tip and surface with a subtip resolution are discussed. PMID- 11419875 TI - Spatial resolution and energy filtering of backscattered electron images in scanning electron microscopy. AB - Numerical simulations of energy filtering effects on backscattered electron images of semiconductor multilayers are reported. The theoretical investigation has been performed for a wide range of energies, 1-40 keV, and for beam incidence angles between 90 degrees (normal incidence) and 20 degrees. Quite a general purpose of this research concerns the investigation of the optimum energy conditions and of their implications. It will be shown that the optimum energy defines an operating context suitable to ensure a compositional contrast enhancement; i.e. a minimum threshold current and a maximum resolution, without energy filtering, independent of the beam incidence angle. This optimum energy, depending on the specimen and its details, is, however, of the order of a few keV or less for specimen details having a size of the order of few nm. When the performance of the electron gun does not allow to work at low energy it is necessary to operate at an energy higher than the optimum one, the energy filtering can produce positive effects. Yet in those circumstances there is an optimum energy loss window suitable to minimise the threshold current. It spreads from 10-30%, depending on the primary energy and size of the compositional detail, for normal incidence, to a few per cent for high incidence angles and high energy. The simulation results for these last conditions are in agreement with the well-known experimental results obtained with the low-loss methods. PMID- 11419876 TI - A high-resolution serial sectioning specimen preparation technique for application to electron backscatter diffraction. AB - A high-resolution serial sectioning specimen preparation technique is described for acquisition of electron backscatter diffraction (EBSD) data. The primary objective is to develop a method to reproducibly remove a controlled thickness of material from a polycrystalline Ta sample while producing quality surfaces for EBSD orientation imaging. This is integrated with the ability to accurately measure the amount of material removed with each iteration and experimentally register the ensuing EBSD scans. To facilitate enhanced accuracy of this method, a metrology device containing high-precision etching patterns is fabricated using standard lithographic techniques. This metrology device allows for the sub-micron measurement of the serial section slice thickness and approximately 1 microm registration accuracy of each EBSD scan. PMID- 11419877 TI - Phase retrieval and aberration correction in the presence of vortices in high resolution transmission electron microscopy. AB - We discuss phase retrieval and the correction of images for aberrations, in particular defocus and spherical aberration, in high-resolution transmission electron microscopy. Non-interferometric phase retrieval requires at least two intensity measurements in different planes. Vortices in the phase may occur in the image plane or the other planes involved in the phase retrieval. We discuss the performance of various methods of phase retrieval in that case. After retrieval of the phase, the aberrations can be corrected in the Fraunhofer diffraction space (the wave function in the diffraction space is related to that in the image space by a Fourier transform). The aberration-corrected image is obtained from the aberration-corrected wave function in the diffraction plane by inverse Fourier transformation. PMID- 11419878 TI - Development of a quantitative energy filtering TEM method to study a reactive NiO/80Ni20Fe interface. AB - Energy filtering TEM (EFTEM) has been performed on an annealed NiO/80Ni20Fe interface. Chemical maps have been calculated using the three-window technique on the O-K, Fe-L23 and Ni-L23 edges. In this paper we show that relative quantification can be made using reference areas on the images taken from part to part of the interface in well-known composition areas. Partial cross-section ratios sigmaNi(20 eV)/sigmaO(20 eV) and sigmaNi(20 eV)/sigmaFe(20 eV) have been then measured and used to extract the chemical composition of the reactive NiO/80Ni20Fe interface. Fe, Ni and O composition profiles across the interface have been obtained showing the diffusion process which has occurred during annealing. The reliability of the EFTEM measurements has been checked by EEL spectroscopy on the same sample as well as on a reference geological oxide sample (Trevorit: NiFe2O4). The partial cross-section ratios obtained with the two methods (EFTEM and EELS) are in good agreement. PMID- 11419879 TI - Physician-performed point-of-care echocardiography using a laptop platform compared with physical examination in the cardiovascular patient. AB - OBJECTIVES: The purpose of this study was to compare the results of physical examinations (PEs) performed by board-certified cardiologists with the results of point-of-care (POC) echocardiography in a group of patients with cardiovascular disease. BACKGROUND: Although cardiovascular PE is crucial in the evaluation of patients with suspected heart disease, the skills required to diagnose abnormal cardiovascular findings have been declining. Echocardiography is a powerful noninvasive cardiovascular diagnostic tool; however, echocardiographic evaluation of patients is not performed at the time of patient encounter (POC echocardiography), beacuse current platforms are cumbersome and expensive for individual physician use. The development of miniaturized echocardiographic equipment has the potential to overcome some of these limitations. METHODS: Thirty-six subjects had a complete cardiovascular examination by four board certified cardiologists. The physicians subsequently imaged each patient using a miniaturized echocardiographic platform. The yield of PE and POC echocardiography were compared using a complete echocardiographic study as the gold standard, performed on an upper-end platform. RESULTS: Cardiac examination failed to detect 59% of the overall cardiovascular findings. Physician-performed echocardiography with the prototype device missed 29% of the overall cardiovascular pathology. When considering only the major cardiovascular findings, the cardiologists' PEs still failed to correctly detect 43%. Point-of-care echocardiography reduced this to 21% without significant interphysician variation. CONCLUSIONS: Point-of-care echocardiography using a miniaturized echocardiographic platform substantially improved the detection of important cardiovascular pathology compared with PE. Use of this device by a cardiovascular specialist with training in echocardiography as a routine adjunct to PE appears to be useful. PMID- 11419880 TI - How useful is hand-carried bedside echocardiography in critically ill patients? AB - OBJECTIVES: The study compared a hand-carried echocardiography (HC) device with standard echocardiography (SE) in critically ill patients. BACKGROUND: Recently, small HC devices have been introduced, and early reports showed a good correlation with SE. METHODS: We used HC (SonoSite, Bothell, Washington) echocardiography to evaluate critically ill patients, and we compared the results with SE obtained with state-of-the-art equipment (Sonos 5500, Hewlett-Packard, Andover, Massachusetts). Each of 80 critically ill patients was studied twice (HC and SE). The studies were done and interpreted separately in blinded fashion. RESULTS: The HC device missed a clinical finding related to the reason for referral in 31% of patients. In 19% of patients a clinically important finding separate from the indication for echocardiography was also missed. The total number of patients with one or more missed findings was 36 (45%). Findings were missed by HC for several reasons. First, HC does not contain spectral Doppler, electrocardiographic, or M-mode capabilities. Two-dimensional imaging is superior on SE, with improved image processing. In addition, although HC does contain color power Doppler, it does not have true color flow Doppler imaging. Therefore, HC often failed to detect or accurately quantify valvular regurgitation. CONCLUSIONS: Although the HC device was able to provide important anatomic information, the device falls far short of SE in the evaluation of critically ill patients. PMID- 11419881 TI - Hand-held echocardiography: revolution or hassle? PMID- 11419882 TI - The deceleration [correction of declaration] time of pulmonary venous diastolic flow is more accurate than the pulmonary artery occlusion pressure in predicting left atrial pressure. AB - OBJECTIVES: This study compared a prediction of mean left atrial pressure (P(LA)) ascertained by Doppler echocardiography of pulmonary venous flow (PVF), with predicted P(LA) using the pulmonary artery occlusion pressure (P(PAO)). BACKGROUND: In select patient groups, PVF variables correlate with P(PAO)) an indirect measure of P(LA). METHODS: In 93 patients undergoing cardiac surgery, we recorded with transesophageal echocardiography mitral valve early (E) and late (A) wave velocities, deceleration time (DT) of E (DT(E)), and pulmonary vein systolic (S) and diastolic (D) wave velocities, DT of D (DT(D)) and systolic fraction. The P(PAO) was measured using a pulmonary artery catheter zeroed to midaxillary level. A further catheter was held at midatrial level to zero a transducer and was then inserted into the left atrium. A prediction rule for P(LA) from DT(D) was developed in 50 patients and applied prospectively to estimate P(LA) in 43 patients. RESULTS: A close correlation (r = -0.92) was found between P(LA) and DT(D). Systolic fraction (r = -0.63), DT(E) (r = -0.61), D wave (r = 0.57), E wave (r = 0.52), and E/A ratio (r = 0.13) correlated less closely with P(LA). The mean difference between predicted and measured P(LA) was 0.58 mm Hg for DT(D) method and 1.72 mm Hg for P(PAO), with limits of agreement (mean +/- 2 SE) of -2.94 to 4.10 mm Hg and -2.48 to 5.92 mm Hg, respectively. A DT(D)) of <175 ms had 100% sensitivity and 94% specificity for a P(LA) of >17 mm Hg. CONCLUSIONS: Deceleration time of pulmonary vein diastolic wave is more accurate than P(PAO) in estimating left atrial pressure in cardiac surgical patients. PMID- 11419883 TI - Transesophageal magnetic resonance imaging of the aortic arch and descending thoracic aorta in patients with aortic atherosclerosis. AB - OBJECTIVES: We sought to determine the feasibility and potential of transesophageal magnetic resonance imaging (TEMRI) for quantifying atherosclerotic plaque burden in the aortic arch and descending thoracic aorta in comparison with transesophageal echocardiography (TEE). BACKGROUND: Improved morphologic assessment of atherosclerotic plaque features in vivo is of interest because of the potential for improved understanding of the pathophysiology of plaque vulnerability to rupture and progression to clinical events. Magnetic resonance imaging (MRI) is well suited for atherosclerotic plaque imaging. Performing MRI using a radio frequency (RF) receiver probe placed near the region of interest improves the signal-to-noise ratio (SNR). METHODS: High-resolution images of the thoracic aortic wall were obtained by TEMRI in 22 subjects (8 normals, 14 with aortic atherosclerosis). In nine subjects, we compared aortic wall thickness and circumferential extent of atherosclerotic plaque measured by TEMRI versus TEE using a Bland-Altman analysis. Additional studies were performed in a human cadaver with pathology as an independent gold standard for assessment of atherosclerosis. RESULTS: In clinical and experimental studies, we found similar measurements for aortic plaque thickness but a relative underestimation of circumferential extent of atherosclerosis by TEE (p = 0.001), due in large part to the lower SNR in the near field. CONCLUSIONS: Using TEMRI allows for quantitative assessment of thoracic aortic atherosclerotic plaque burden. This technique provides good SNR in the near field, which makes it a promising approach for detailed characterization of aortic plaque burden. PMID- 11419884 TI - Effects of low-dose aspirin on serum C-reactive protein and thromboxane B2 concentrations: a placebo-controlled study using a highly sensitive C-reactive protein assay. AB - OBJECTIVES: We performed a placebo-controlled study to evaluate the effect of low dose aspirin on serum C-reactive protein (CRP) levels. BACKGROUND: Elevated circulating concentrations of CRP, an inflammatory marker, increase the risk of thrombotic cardiovascular diseases such as myocardial infarction (MI). Moreover, low-dose aspirin therapy has been reported to be more effective in preventing MI in men with higher CRP levels than it is in those with lower levels, raising the possibility that aspirin prevents thrombosis by reducing vascular inflammation. The effect of low-dose aspirin therapy on serum CRP levels in men has been addressed recently, but the results of the two studies conflict. METHODS: Effects of aspirin (81 mg every day or 325, 81 or 40 mg every-third-day given for 31 days) on serum CRP, using a highly-sensitive assay, and on serum platelet-cyclo oxygenase (COX)-1-derived thromboxane (Tx) B2 concentrations were studied simultaneously in 57 healthy volunteers (30 men and 27 women). RESULTS: Trough platelet COX-1-derived serum Tx B2 concentrations decreased by 100% with daily aspirin and by 90%, 84% and 78% with 325, 81 and 40 mg aspirin every-third-day (p < 0.001). However, there were no significant changes in serum CRP levels from baseline with daily low-dose aspirin therapy, with any of the every-third-day aspirin regimens or with placebo treatment. CONCLUSIONS: Low doses of aspirin that markedly inhibit platelet COX-1 activity, as manifested by a profound decline in platelet-derived serum Tx B2 concentrations, have no detectable effect on serum CRP levels in healthy men and women. PMID- 11419885 TI - Randomized comparison of a strategy of predischarge coronary angiography versus exercise testing in low-risk patients in a chest pain unit: in-hospital and long term outcomes. AB - OBJECTIVES: This randomized trial compared a strategy of predischarge coronary angiography (CA) with exercise treadmill testing (ETT) in low-risk patients in the chest pain unit (CPU) to reduce repeat emergency department (ED) visits and to identify additional coronary artery disease (CAD). BACKGROUND: Patients with chest pain and normal electrocardiograms (ECGs) have a low likelihood of CAD and a favorable prognosis, but they often seek repeat evaluations in EDs. Remaining uncertainty regarding their symptoms and diagnosis may cause much of this recidivism. METHODS: A total of 248 patients with no ischemic ECG changes triaged to a CPU were randomized to CA (n = 123) or ETT (n = 125). All patients had a probability of myocardial infarction < or =7% according to the Goldman algorithm, no biochemical evidence of infarction, the ability to exercise and no previous documented CAD. Patients were followed up for > or =1 year and surveyed regarding their chest pain self-perception and utility of the index evaluation. RESULTS: Coronary angiography showed disease (> or =50% stenosis) in 19% and ETT was positive in 7% of the patients (p = 0.01). During follow-up (374+/-61 days), patients with a negative CA had fewer returns to the ED (10% vs. 30%, p = 0.0008) and hospital admissions (3% vs. 16%, p = 0.003), compared with patients with a negative/nondiagnostic ETT. The latter group was more likely to consider their pain as cardiac-related (15% vs. 7%), to be unsure about its etiology (38% vs. 26%) and to judge their evaluation as not useful (39% vs. 15%) (p < 0.01 for all comparisons). CONCLUSIONS: In low-risk patients in the CPU, a strategy of CA detects more CAD than ETT, reduces long-term ED and hospital utilization and yields better patient satisfaction and understanding of their condition. PMID- 11419886 TI - Defining the role of chest pain units. PMID- 11419887 TI - Clinical risk stratification correlates with the angiographic extent of coronary artery disease in unstable angina. AB - OBJECTIVES: We sought to determine whether clinical risk stratification correlates with the angiographic extent of coronary artery disease (CAD) in patient with unstable angina. BACKGROUND: The Agency for Health Care Policy and Research (AHCPR) guidelines stratify patients with unstable angina according to short-term risk of myocardial infarction or death. Whether these guidelines are useful in predicting the extent of CAD is unknown. METHODS: All residents of Olmsted County, Minnesota, undergoing emergency department evaluation from January 1, 1985 through December 31, 1992 for unstable angina without a history of prior coronary artery bypass grafting, and who underwent early angiography (within seven days of presentation) were classified into low, intermediate and high risk subgroups based on AHCPR criteria. RESULTS: Seven hundred ninety-five patients underwent early angiography: 159 high risk, 572 intermediate risk and 64 low risk patients. Logistic regression analysis demonstrated that low risk patients had a greater likelihood of normal or mild CAD relative to intermediate risk (odds ratio [OR], 4.67; 95% confidence interval [CI], 2.70-8.06; p < 0.001) and high risk (OR, 11.1; 95% CI, 5.71-22.2; p < 0.001). Significant 1-, 2-, 3 vessel coronary disease or left main coronary disease was more likely in high relative to low risk (OR, 8.09; 95% CI, 4.22-15.5; p < 0.001), intermediate relative to low risk (OR, 4.11; 95% CI, 2.34-7.22; p < 0.001), and high relative to intermediate risk (OR, 1.97; 95% CI, 1.31-2.96; p = 0.0012). CONCLUSIONS: Among patients with unstable angina undergoing early coronary angiography, risk stratification according to the AHCPR guidelines correlates with the angiographic extent of CAD. PMID- 11419888 TI - Long-term mortality benefit with abciximab in patients undergoing percutaneous coronary intervention. AB - OBJECTIVES: The goal of this study was to test: 1) if platelet glycoprotein IIb/IIIa (GP IIb/IIIa) blockade with abciximab bolus plus 12-h infusion reduces mortality after percutaneous coronary intervention (PCI); 2) if prevention of early myocardial infarction (MI) after PCI is a mechanism for reducing mortality; and 3) for risk factors for mortality after PCI. BACKGROUND: Studies of PCI suggest that MI after intervention is predictive of mortality. Abciximab, a platelet GP IIb/IIIa receptor inhibitor, has consistently reduced the incidence of MI among PCI patients in several trials. The presumed mechanism is prevention of platelet thrombus associated with vessel wall injury and downstream embolization into the microcirculation. METHODS: In eight trials, 5,154 patients were randomized to a regimen comprising conventional therapy plus a bolus of abciximab within 1 h before PCI followed by a 12-h infusion; 4,136 controls were randomized to conventional therapy alone. Patient follow-up from six months to three years was available. Survival differences are examined using proportional hazards regression and survival curves. RESULTS: A hazard ratio of 0.71 (95% confidence interval 0.57 to 0.89; p = 0.003) suggests a mortality benefit with abciximab. The absolute reduction in mortality was estimated to be 0.5% through 30 days, 0.7% through six months, 0.9% through one year and 1.8% through three years. Early MI explained 18% of the observed mortality benefit at one year. Multivariate regression suggests that patients with advanced cardiovascular disease may derive the greatest mortality benefit from abciximab. CONCLUSIONS: The evidence from 9,290 randomized PCI patients shows a mortality benefit provided by abciximab bolus plus 12-h infusion. PMID- 11419889 TI - Incidence of thrombotic occlusion and major adverse cardiac events between two and four weeks after coronary stent placement: analysis of 5,678 patients with a four-week ticlopidine regimen. AB - OBJECTIVES: We attempted to make a comprehensive assessment of the risk of stent failure (death, myocardial infarction or angiographically documented occlusion), differentiating early (first and second weeks) and late (third and fourth weeks) events. BACKGROUND: The risk of stent failure decreases rapidly within the first week. It has been suggested that the risk rate for late events is close to 0% and that the thienopyridine regimen (ticlopidine or clopidogrel) could be safely reduced from four to two weeks, minimizing the risk of hematological complications. METHODS: We analyzed 5,678 patients with successful coronary stent placement and a four-week ticlopidine regimen. RESULTS: The rate of stent failure was 2.5% at four weeks, with 112 early (2.0%) and 30 late events (0.5%). Multivariate analysis identified different risk factors for early versus late events. While variables on stenosis severity and procedural results that can be influenced by the operator were identified as independent risk factors for early events (percent stenosis before and after the procedure, residual dissection, length of stented segment), more clinical variables were associated with late events (age, reduced left ventricular function, systemic hypertension as a protective factor). The late-event rate was <0.1% in the absence of these factors, but it was 2.5% with all three risk factors present. CONCLUSIONS: The risk of late stent failure is low with a four-week ticlopidine regimen. However, high-risk subgroups have a risk of 2.5%. As this rate is presumably higher if thienopyridines are discontinued after two weeks, these data suggest that a risk stratification to a two- or four-week regimen is preferable to a general reduction. PMID- 11419890 TI - Emergency stenting to treat neurological complications occurring after carotid endarterectomy. AB - OBJECTIVES: The purpose of this study was to assess the efficacy of emergency stent implantation for the treatment of perioperative stroke after carotid endarterectomy (CEA). BACKGROUND: Carotid endarterectomy has been proven safe and effective in reducing the risk of stroke in symptomatic and asymptomatic patients with >60% carotid artery stenosis. However, perioperative stroke has been reported in 1.5% to 9% of CEA cases. The management of such a complication is challenging. Recently, percutaneous transluminal carotid angioplasty with stent deployment has emerged as a valuable and alternative strategy for the treatment of carotid artery disease. METHODS: Between April 1998 and February 2000, 18 of the 995 patients (1.8%) who had CEA in our institution experienced perioperative major or minor neurological complications. Of these, 13 patients underwent emergency carotid angiogram and eventual stent implantation, whereas the remaining five had surgery re-exploration. RESULTS: Carotid angiogram was performed within 20+/-10 min and revealed vessel flow-limiting dissection (five cases) or thrombosis (eight cases). Percutaneous transluminal carotid angioplasty with direct stenting (self-expandable stent) was performed in all 13 cases. Angiographic success was 100%. Complete remission of neurological symptoms occurred in 11 of the 13 patients treated by stent implantation and in one of the five patients treated by surgical re-exploration (p = 0.024). CONCLUSIONS: Stent implantation seems to be a safe and effective strategy in the treatment of perioperative stroke complicating CEA, especially when carotid dissection represents the main anatomic problem. PMID- 11419891 TI - Skeletal muscle mass independently predicts peak oxygen consumption and ventilatory response during exercise in noncachectic patients with chronic heart failure. AB - OBJECTIVES: We sought to assess whether skeletal muscle mass might be a predictor of peak oxygen consumption (Vo2) and relation of the ventilation to carbon dioxide production (VE/VCo2) slope in patients with chronic heart failure (CHF) independent of clinical conditions, neurohormonal activation and resting hemodynamics. BACKGROUND: A variety of abnormalities characterize skeletal muscle and contribute to exercise intolerance in patients with CHF. Skeletal muscle mass is a determinant of peak Vo2 both in healthy patients and in patients with CHF, but there are no reports on the independent predictive value of this parameter, which can be measured with great accuracy by whole-body dual energy X-ray absorptiometry (DEXA). The influence of skeletal muscle mass on VE/VCo2 slope is not known either. METHODS: We prospectively evaluated 120 consecutive noncachectic patients with CHF. Every patient underwent a cardiopulmonary exercise test, an echo-Doppler examination and an evaluation of neurohormonal activation and body composition as assessed by DEXA. RESULTS: At the univariate analysis, New York Heart Association (NYHA) class (p < 0.0001), age (p < 0.0001), male gender (p < 0.0001) and plasma renin (p < 0.0001) significantly related with peak Vo2. There was a significant correlation between lean mass and absolute peak Vo2 (r = 0.70, p < 0.0001) and VE/VCo2 slope (r = -0.27; p < 0.01). At the multivariate analysis, lean mass predicted peak Vo2 and VE/VCo2 slope independently of NYHA functional class, age, gender, neurohormonal activation and resting hemodynamics. CONCLUSIONS: Skeletal muscle mass is an independent predictor of peak Vo2 and VE/VCo2 slope in stable noncachectic patients with CHF. Future studies will determine whether an increase in skeletal muscle mass in the individual patient might result in an improvement in parameters of exercise capacity. PMID- 11419892 TI - Relationship between tumor necrosis factor-alpha production and oxidative stress in the failing hearts of patients with dilated cardiomyopathy. AB - OBJECTIVES: This study evaluated oxidative stress in the failing ventricle in patients with dilated cardiomyopathy (DCM). BACKGROUND: Oxidative stress appears to increase in the failing myocardium and may contribute to ventricular dysfunction in patients with DCM. Tumor necrosis factor-alpha (TNF-alpha), which is expressed in the failing heart, may stimulate oxidative stress. METHODS: We measured plasma oxidized low density lipoprotein (oxLDL) by sandwich enzyme linked immunosorbent assay using specific antibodies against oxLDL in the aortic root (AO) and the coronary sinus (CS) in control subjects (n = 8) and in 22 patients with DCM and mild congestive heart failure. We also measured the plasma levels of TNF-alpha and angiotensin II. RESULTS: There was no difference in oxLDL between the AO and CS in control subjects. In contrast, plasma oxLDL was significantly higher in the CS than the AO in patients with DCM, suggesting that the transcardiac gradient ofoxLDL reflects oxidative stress in the failing heart in these patients. Plasma TNF-alpha levels were significantly higher in the CS than the AO with a significant positive correlation of the transcardiac gradient of TNF-alpha and the transcardiac gradient of oxLDL. Moreover, a significant negative correlation existed between the transcardiac gradient of oxLDL and left ventricular ejection fraction. The transcardiac gradient of plasma oxLDL was significantly lower in 6 patients who received carvedilol than in 16 patients who did not receive carvedilol. CONCLUSIONS: These findings indicate that the transcardiac gradient of oxLDL may be a marker of oxidative stress in the heart and that left ventricular dysfunction may be partly due to the oxidative stress in patients with DCM. In addition, TNF-alpha may stimulate oxidative stress in the failing heart in patients with DCM. PMID- 11419893 TI - Left ventricular dysfunction after long-term right ventricular apical pacing in the young. AB - OBJECTIVES: The goal of this study was to assess long-term global left ventricular (LV) function in patients paced from the right ventricular (RV) apex at a young age. BACKGROUND: Ventricular contraction asynchrony with short-term RV apical pacing has been associated with reduced LV pump function and relaxation. The long-term effect of RV apical pacing on global LV function in the young remains unknown. METHODS: Twenty-four patients with normal segmental anatomy paced from the RV apex (follow-up 1 to 19 years) underwent noninvasive assessment of global LV function with automated border detection echocardiography-derived fractional area of change (FAC), coupled with the Doppler index of myocardial performance (MPI). Data were analyzed from 24 RV-paced patients (mean follow-up 9.5 years, age 19 years, body surface area [BSA] 1.6 m2, QRS duration 140 ms) and compared with 33 age- and BSA-matched control subjects (age 16.4 years, BSA 1.6 m2). Multiple linear regression analysis was performed to identify patient variables that can affect these indexes of LV function. RESULTS: Assessment of LV function (median follow-up 10 years) in 24 paced patients demonstrated impaired area- and Doppler flow-derived indexes of LV systolic and diastolic function, compared with those indexes of control subjects (FAC: 52% vs. 60%, p < 0.01; MPI: 0.46 vs. 0.34, p < 0.01). Paced QRS interval and age were found to significantly influence global LV contraction in these patients (R2 = 0.4, p < 0.05). CONCLUSIONS: In the presence of impaired LV function with long-term RV apical pacing, alternative sites of ventricular pacing that simulate normal biventricular electrical activation should be explored to preserve function in pediatric patients in need of long-term pacing. PMID- 11419894 TI - Aortic stenosis with severe left ventricular dysfunction and low transvalvular pressure gradients: risk stratification by low-dose dobutamine echocardiography. AB - OBJECTIVES: We sought to assess risk stratification by using dobutamine stress echocardiography (DSE) in patients with aortic stenosis (AS) and severe left ventricular (LV) dysfunction. BACKGROUND: Few data are available on risk stratification for valve replacement in patients with AS, LV dysfunction and low transvalvular gradients. METHODS: Low-dose DSE was performed in 45 patients (16 women and 29 men; median [quartile range] age in years: 75 [69 to 79]; left ventricular ejection fraction: 0.29 [0.23 to 0.32]; aortic valve area [cm2]: 0.7 [0.5 to 0.8]; mean transaortic gradient [mm Hg]: 26 [21 to 33]). Patients were classified into two groups: group I (n = 32, LV contractile reserve on DSE) and group II (n = 13, no contractile reserve). Valve replacement was performed in 24 and 6 patients in groups I and II, respectively. RESULTS: Perioperative mortality was 8% in group I and 50% in group II (p = 0.014). Survival at five years after the operation was 88% in group I. Compared with medical therapy, valve surgery was associated with better long-term survival in group I (hazard ratio for death [HR-D] 0.13, 95% confidence interval [CI] 0.002 to 0.49) and reduced survival in group II (HR-D 19.6, 95% CI 2.7 to 142). The effect of valve surgery on survival remained significant in both groups after adjustment for age, diabetes, respiratory disease and hypertension. Medical therapy had the same effect in both groups. CONCLUSIONS: In patients with AS, LV dysfunction and low transvalvular gradients, contractile reserve on DSE is associated with a low operative risk and good long-term prognosis after valve surgery. In contrast, operative mortality remains high in the absence of contractile reserve. PMID- 11419895 TI - Right ventricular form and function after percutaneous atrial septal defect device closure. AB - OBJECTIVES: We sought to assess the right heart's response to percutaneous device closure of moderate sized atrial septal defects (ASDs) in adults over a one-year follow-up period. BACKGROUND: Percutaneous ASD device closure is a safe and effective means of reducing or eliminating interatrial shunting. The response of the adult's right heart to device closure is incompletely understood. METHODS: Forty consecutive patients had 40 device implantations (32 with the CardioSeal implant and 8 with the Amplatzer device). The patients were assessed with echocardiography, chest radiography and electrocardiography before the procedure and at 1, 6 and 12 months. RESULTS: The mean ASD size was 13+/-4 mm, and the device size ranged from 33 to 40 mm for CardioSeal and 12 to 36 mm for Amplatzer. At one month, heart size (49% vs. 46%), four-chamber right ventricular (RV) size (45 vs. 41 mm), paradoxical septal motion (60% vs. 5%), QRS duration (125 vs. 119 ms), PR interval (181 vs. 155 ms) and echocardiographically determined pulmonary artery systolic pressure decreased significantly and was maintained at 12-month follow-up. At six months, right atrial length decreased from 50 to 47 mm. At one year, 29% of patients had persistent RV enlargement. CONCLUSIONS: Right heart morphology undergoes rapid improvement within one month of defect closure, with associated mechanoelectrical benefit. A small number of patients had persistent RV enlargement or pulmonary hypertension, or both, at one year. Our data support the application of transcatheter methods in achieving excellent hemodynamic and anatomic outcomes. PMID- 11419896 TI - Association of chromosome 22q11 deletion with isolated anomalies of aortic arch laterality and branching. AB - OBJECTIVES: The purpose of this study was to determine the frequency of chromosome 22q11 deletions in patients with isolated anomalies of the aortic arch and its branches. BACKGROUND: Chromosome 22q11 deletions are often present in patients with certain forms of congenital cardiovascular disease, including tetralogy of Fallot, truncus arteriosus and interruption of the aortic arch. Among patients with these anomalies, chromosome 22q11 deletion is more common in those with abnormal aortic arch laterality or branching. METHODS: We studied 66 patients with isolated anomalies of the aortic arch and no associated intracardiac defects for deletions within chromosome 22q11, using fluorescence in situ hybridization with the cosmid probe N25 (D22S75). Arch anomalies included: double aortic arch (n = 22); right aortic arch with aberrant left subclavian artery (n = 28); right aortic arch with mirror-image branching and a vascular ring formed by a left-sided ductus from the descending aorta (n = 5); right aortic arch with mirror-image branching and no vascular ring (n = 4); and left aortic arch with aberrant right subclavian artery (n = 7). In addition, four patients had a cervical aortic arch, four had aortic coarctation and six had hypoplasia/atresia of the proximal pulmonary arteries. RESULTS: Chromosome 22q11 deletions were found in 16 patients (24%) across the full spectrum of anomalies studied. Among the morphologic variables analyzed, only hypoplasia/atresia of the proximal pulmonary arteries correlated with the deletion (p = 0.03). Among patients with a double arch, the frequency of chromosome 22q11 deletion was higher in those with an atretic minor arch than it was in those with a patent minor arch (p = 0.02). CONCLUSIONS: Chromosome 22q11 deletion is associated with isolated anomalies of laterality or branching of the aortic arch in 24% of cases in our series. These findings should alert the clinician to consider deletion screening in patients with isolated anomalies of the aortic arch. PMID- 11419897 TI - Myocardial flow reserve in patients with a systemic right ventricle after atrial switch repair. AB - OBJECTIVES: The purpose of this study was to assess myocardial blood flow (MBF) and flow reserve in systemic right ventricles (RV) in long-term survivors of the Mustard operation. BACKGROUND: There is a high prevalence of systemic RV dysfunction and impaired exercise performance in long-term survivors of the Mustard operation. A mismatch between myocardial blood supply and systemic ventricular work demand has been proposed as a potential mechanism. METHODS: We assessed MBF at rest and during intravenous adenosine hyperemia in 11 long-term survivors of a Mustard repair (age 18+/-5 years, median age at repair 0.7 years, follow-up after repair 17+/-5 years) and 13 healthy control subjects (age 23+/-7 years), using N-13 ammonia and positron emission tomography imaging. RESULTS: There was no difference in basal MBF between the systemic RV of survivors of the Mustard operation and the systemic left ventricle (LV) of healthy control subjects (0.80+/-0.19 vs. 0.74+/-0.15 ml/g/min, respectively, p = NS). However, the hyperemic flows were significantly lower in systemic RVs than they were in systemic LVs (2.34+/-0.0.69 vs. 3.44+/-0.62 ml/g/min respectively, p < 0.01). As a result, myocardial flow reserve was lower in systemic RVs than it was in systemic LVs (2.93+/-0.63 vs. 4.74+/-1.09, respectively, p < 0.01). CONCLUSIONS: Myocardial flow reserve is impaired in systemic RVs in survivors of the Mustard operation. This may contribute to systemic ventricular dysfunction in these patients. PMID- 11419898 TI - Effect of human recombinant vascular endothelial growth factor165 on progression of atherosclerotic plaque. AB - OBJECTIVES: This study was designed to evaluate the impact of recombinant human vascular endothelial growth factor165 (rhVEGF) on atherosclerotic plaque progression. BACKGROUND: Therapeutic angiogenesis represents a promising treatment for ischemic diseases. However, angiogenesis may impact atherosclerosis. METHODS: Albumin or rhVEGF was administered by a single intramuscular injection (2 microg/kg body weight) to New Zealand White rabbits fed with a 0.25% cholesterol diet beginning three weeks before therapy. Subsets of rabbits from each group underwent perfusion-fixation and harvesting of the thoracic aorta for morphometric and immunohistochemical analyses at 7 or 21 days. RESULTS: The mean plaque area was 15.75+/-2.28% and 22.00+/-3.24% with VEGF and 0.67+/-0.22% and 1.17+/-0.34% with albumin at 7 and 21 days, respectively. The plaque circumference was 13.00+/-2.58% and 23.75+/-2.86% with VEGF and 2.50+/ 0.65% and 6.25+/-1.88% with albumin at 7 and 21 days, respectively. The maximal plaque thickness was 0.11+/-0.002 and 0.15+/-0.007 mm with VEGF and 0.04+/-0.009 and 0.07+/-0.003 mm with albumin at 7 and 21 days, respectively. The endothelial density (reported as percent total plaque area) was 31.75+/-4.42% and 63.00+/ 8.45% with VEGF and 7.75+/-1.65% and 12.75+/-1.93% with albumin at 7 and 21 days, respectively. The macrophage density was 4.5+/-0.86 and 19.25+/-1.54 with VEGF and 4.26+/-0.75 and 6.00+/-1.08 with albumin at 7 and 21 days, respectively. CONCLUSIONS: Recombinant human VEGF increases the rate and degree of atherosclerotic plaque formation in the thoracic aorta in a cholesterol-fed rabbit model. PMID- 11419899 TI - Vascular endothelial growth factor-induced angiogenesis: crouching tiger or hidden dragon? PMID- 11419900 TI - Evaluation of the role of I(KACh) in atrial fibrillation using a mouse knockout model. AB - OBJECTIVES: We sought to study the role of I(KACh) in atrial fibrillation (AF) and the potential electrophysiologic effects of a specific I(KACh) antagonist. BACKGROUND: I(KACh) mediates much of the cardiac responses to vagal stimulation. Vagal stimulation predisposes to AF, but the specific role of I(KACh) in the generation of AF and the electrophysiologic effects of specific I(KACh) blockade have not been studied. METHODS: Adult wild-type (WT) and I(KACh)-deficient knockout (KO) mice were studied in the absence and presence of the muscarinic receptor agonist carbachol. The electrophysiologic features of KO mice were compared with those of WT mice to assess the potential effects of a specific I(KACh) antagonist. RESULTS: Atrial fibrillation lasting for a mean of 5.7+/-11 min was initiated in 10 of 14 WT mice in the presence of carbachol, but not in the absence of carbachol. Atrial arrhythmia could not be induced in KO mice. Ventricular tachyarrhythmia could not be induced in either type of mouse. Sinus node recovery times after carbachol and sinus cycle lengths were shorter and ventricular effective refractory periods were greater in KO mice than in WT mice. There was no significant difference between KO and WT mice in AV node function. CONCLUSIONS: Activation of I(KACh) predisposed to AF and lack of I(KACh) prevented AF. It is likely that I(KACh) plays a crucial role in the generation of AF in mice. Specific I(KACh) blockers might be useful for the treatment of AF without significant adverse effects on the atrioventricular node or the ventricles. PMID- 11419901 TI - Left ventricular diastolic dysfunction during demand ischemia: rigor underlies increased stiffness without calcium-mediated tension. Amelioration by glycolytic substrate. AB - OBJECTIVES: The goal of this study was to determine the subcellular mechanism(s) underlying increased left ventricular (LV) diastolic chamber stiffness (DCS) during angina (demand ischemia). BACKGROUND: Increased DCS may result from increased diastolic myocyte calcium concentration and/or rigor. Therefore, we assessed the effects of direct alterations of both calcium-activated tension and high-energy phosphates on increased DCS. METHODS: Demand ischemia was reproduced in isolated, isovolumic, red-cell perfused rabbit hearts by imposing low-flow ischemia and pacing tachycardia. This resulted in increased DCS. Interventions were performed after LV end-diastolic pressure had increased approximately 7 mm Hg. Initially, to determine the effects of altered calcium concentration or myofilament calcium responsiveness, hearts received either: 1) 5 or 14 mmol/L calcium chloride; 2) 8 mmol/L egtazic acid; 3) 5 mmol/L butane-dione-monoxime (BDM); or 4) 50 mmol/L ammonium chloride (NH4Cl). Then, to assess the contribution of decreased high-energy phosphate supply, hearts received 5) glucose (25 mmol/L) and insulin (400 microU/ml). RESULTS: 1) Calcium chloride, 5 and 14 mmol/L, increased LV systolic pressure by 42% and 70%, respectively (p < 0.001), indicating increased calcium-activated tension, but did not further increase DCS, implying intact diastolic calcium resequestration. 2) Egtazic acid reduced LV systolic pressure by 30% (p < 0.001), indicating reduced intracellular calcium, but failed to reduce increased DCS. 3) Butane-dione-monoxime and NH4Cl chloride affected contractile function (i.e., a calcium-driven force) but did not alter increased DCS. 4) Glucose and insulin, which increase high-energy phosphates during ischemia, reduced increased DCS by 50% (p < 0.001). CONCLUSIONS: Increased DCS during demand ischemia was insensitive to maneuvers altering intracellular calcium concentration or myofilament calcium responsiveness, that is, evidence against an etiology of calcium-activated tension. In contrast, increased glycolytic substrate ameliorated increased DCS, supporting a primary mechanism of rigor-bond formation. PMID- 11419902 TI - Differential effects of angiotensin AT1 and AT2 receptors on the expression, translation and function of the Na+-H+ exchanger and Na+-HCO3- symporter in the rat heart after myocardial infarction. AB - OBJECTIVES: This study investigated the role of angiotensin receptor subtype 1 (AT1) and angiotensin receptor subtype 2 (AT2) in the regulation of Na+-H+ exchanger (NHE) and Na+-HCO3 symporter (NBC) in the infarcted myocardium. BACKGROUND: The cardiac renin-angiotensin system is activated after myocardial infarction (MI), and both angiotensin AT1 and AT2 receptors are upregulated in the myocardium. METHODS: Na+-H+ exchanger isoform-1 and NBC-1 gene expression were determined by reverse transcription polymerase chain reaction and Northern blot analysis; protein levels by Western blot analysis; and activity by measurement of H+ transport in left ventricular (LV) free wall, interventricular septum (IS) and right ventricle (RV) after induction of MI. Rats were treated with placebo, the angiotensin-converting enzyme inhibitor ramipril (1 mg/kg/day), the AT1 receptor antagonist valsartan (10 mg/kg/day) or the AT2 receptor antagonist PD 123319 (30 mg/kg/day). Treatment was started seven days before surgery. RESULTS: Na+-H+ exchanger isoform-1 and NBC-1 messenger RNA (mRNA) expression and protein levels were increased twofold in the LV free wall after MI, whereas no changes were observed in the IS and RV. Na+-dependent H+ flux was increased in the LV free wall. Ramipril inhibited mRNA and protein upregulation of both transporters. Valsartan inhibited the upregulation of NHE-1 mRNA and protein but had no effect on NBC-1 mRNA expression and translation. In contrast, PD 123319 abolished the upregulation of NBC-1 mRNA and protein but had no effect on NHE-1 upregulation. Ramipril and valsartan prevented post-MI increase in NHE-1 activity, whereas ramipril and PD 123319 decreased NBC-1 activity. CONCLUSIONS: Angiotensin II via its AT1 and AT2 receptors differentially controls transcriptional and translational regulation as well as the activity of NHE-1 and NBC-1 in the ischemic myocardium and contributes to the control of pH regulation in cardiac tissue. PMID- 11419903 TI - President's page: Complementary and alternative medicine: ignore at doctors' and patients' peril. PMID- 11419904 TI - American College of Cardiology/Society for Cardiac Angiography and Interventions Clinical Expert Consensus Document on cardiac catheterization laboratory standards. A report of the American College of Cardiology Task Force on Clinical Expert Consensus Documents. PMID- 11419905 TI - ACC/AHA guidelines of percutaneous coronary interventions (revision of the 1993 PTCA guidelines)--executive summary. A report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (committee to revise the 1993 guidelines for percutaneous transluminal coronary angioplasty). PMID- 11419906 TI - The American College of Cardiology-National Cardiovascular Data Registry (ACC NCDR): building a national clinical data repository. PMID- 11419907 TI - Cell culture conditions affect LPS inducibility of the inflammatory mediators in J774A.1 murine macrophages. AB - Peripheral blood monocytes and tissue macrophages contribute significantly to the inflammatory response. Bacterial lipopolysaccharide (LPS) has profound effects on these cells including, but not limited, to differentiation into macrophages, production of reactive nitrogen and oxygen species and secretion of pro inflammatory cytokines. Herein, we describe a variant of the J774A.1 murine macrophage line that is reversibly resistant to multiple effects of LPS when cultured in different types of media. J774A.1 cells are adherent and spread out when cultured in DMEM/F12; however, when cultured in RPMI 1640, the cells are rounded and relatively non-adherent. Different types of tissue culture plates, sera, and media supplements were not responsible for these changes. We examined LPS-induced reactive nitrogen species using the Greiss reagent. J774A.1 cells cultured in RPMI exhibit a 5-fold increase in nitrites in culture supernatants after LPS stimulation whereas those in DMEM/F12 do not. Zinc staining of total cellular protein of cells in COHLY ET AL. RPMI and DMEM/F12 electrophoresed on a SDS-PAGE showed noticeable banding differences. LPS-induced cytokine gene expression was studied by RT-PCR. LPS induced TNF-alpha, IL-1alpha, IL-1beta, and sIL-1Ra in cells cultured in RPMI but not those cultured in DMEM/F12 with the exception of TNFalpha. This report shows that environmental factors contained in the culture medium alone can reversibly alter the biochemical nature of monocytes and macrophages. PMID- 11419908 TI - The effects of melatonin on humoral immune responses of young and aged rats. AB - The pineal gland with its effects on immune system and free radicals may have a role on aging process. In this study, we investigated the effects of melatonin (10 mg/kg/d, s.c. for 7 days), the main secretion of the pineal gland, on the humoral immune responses of aged and young male Wistar rats. Eighteen aged (28 months old; 6 in control group and 12 in melatonin group) and 25 young (9 months old; 10 in control group and 15 in melatonin group) rats were given 4 x 10(8) sheep erythrocytes i.p. in order to evoke humoral immune responses. After a booster injection at the end of a period of three-weeks following the last melatonin injection, IgM and IgG1 levels were measured. Melatonin was found to increase IgG1 and IgM responses of aged rats when compared to controls (p = 0.049 and p = 0.007), respectively. In the young rats, while the IgG1 levels of control group were significantly higher than that of the melatonin group (p =0.021), IgM levels were not significantly different (p = 0.563). It is concluded that exogenous melatonin may augment the depressed humoral immune responses seen aged rats. PMID- 11419909 TI - The interaction of selective plant lectins with neuraminidase-treated and untreated IgA1 from the sera of IgA nephropathy patients. AB - A study on the binding interaction of lectins from Artocarpus heterophyllus (jacalin), Glycine max and Sambucus nigra with standardised quantity of IgA from the IgA nephropathy patients and normal controls was performed. The Glycine max lectin demonstrated higher affinity towards the serum IgA of IgAN patients as compared to normal controls. However, the affinity binding was lower in cases ofjacalin and the Sambucus nigra lectin. When serum samples were treated with neuraminidase, the differential jacalin affinity binding between IgA1 of patients and normal controls was abrogated. Our data are in support of the view that the O linked oligosaccharide moieties of the patients IgA1 were generally lacking in galactose and sialic acid residues. PMID- 11419910 TI - Induction of T cell responses against autologous ovarian tumors with whole tumor cell lysate-pulsed dendritic cells. AB - The loading of dendritic cells (DCs) with whole tumor cell lysates may circumvent the facts that few tumor-specific antigens have been identified in human solid tumors. The present study was designed to investigate whether ovarian cancer cells lysate-pulsed DCs activate T cell responses against autologous ovarian tumors. Incubation of T cells with autologous tumor cell lysate-pulsed DCs stimulated proliferation of autologous T cells. T cells primed by autologous tumor cell lysate-pulsed DCs showed significant killing activity against autologous tumor cells, which could be blocked by anti-MHC-class-I and anti-CD8 mAb. By contrast, T cells primed by autologous unpulsed DCs alone or tumor lysates alone failed to exhibit significant killing activity. In addition, T cells primed by DCs pulsed with allogeneic tumor cell lysates or with autologous normal cell lysate or by these cell lysates alone did not induce the increase in the autologous tumor killing activity. As additional controls, T cells stimulated with autologous tumor lysate-pulsed DCs express no increase in the lysis of autologous monocytes, allogeneic ovarian tumor cells and other cell lines including K562, Daudi and Molt-4. Furthermore, T cells stimulated with autologous tumor lysate-pulsed DCs could produce the considerable amounts of cytokines such as GM-CSF, TNF-alpha and IFN-gamma. The data in the present study suggest that whole tumor cell lysates-pulsed DCs could activate T cell responses against autologous ovarian tumor cells, and that these pulsed DCs may be used as a new approach for the specific immunotherapy of ovarian cancer patients. PMID- 11419911 TI - Humoral immune-mediated acute, antigen-induced arthritis in rats is suppressed by the inducing antigen administered orally before, but not after, immunization. AB - Acute ovalbumin-induced arthritis (OIA), which is mediated by Arthus reaction to ovalbumin (OVA) in the joint space, can be induced by immunization of rats with OVA followed by the intraarticular injection of OVA. Because oral administration of antigen induces immunological unresponsiveness, we studied for the first time the effects of oral administration of OVA on acute OIA. The oral administration of OVA before immunization significantly suppressed the development of acute OIA, in accordance with decreases in both the anti-OVA IgG antibody production and in vitro lymphocyte proliferative responses to OVA. However, the oral administration of OVA after immunization did not show any decrease in antibody production or in vitro lymphocyte proliferation to OVA, or in the severity of acute OIA. These results indicate that the induction of oral tolerance to OVA in OIA is possible by oral administration of OVA before, but not after, immunization with the antigen. This supports the concept of using antigen feeding as a treatment for certain humoral immune-mediated diseases. PMID- 11419912 TI - A quantitative analysis of CD45Rlow CD4+ T cells in the subarachnoid space of Lewis rats with autoimmune encephalomyelitis. AB - In order to quantify encephalitogenic effector T cells in the subarachnoid space (SAS) and spinal cord afflicted with experimental autoimmune encephalomyelitis (EAE), we immunized Lewis rats using myelin basic protein and complete Freund's adjuvants and analyzed the inflammatory cells by fluorescence-activated cell sorter (FACS) and immunohistochemistry. At the induction stage of EAE, the majority of observed inflammatory cells were determined by immunohistochemistry to be either CD4+ T cells or OX42+ macrophages. Among CD4+ T cells, both CD45R high (OX22+) and CD45R low (OX22-) T cells were found in the SAS, while in the neighboring subpial spinal cord parenchyma, CD45R low (OX22-negative)/ CD4+ T cells predominated. FACS analysis showed that CD45RC low/CD4+ T cells was 83% of total CD4+ T cells in the SAS, while 94% of cells with the same phenotype were found in the parenchyma of rat spinal cords afflicted with EAE. This finding suggests that during the induction stage of EAE, effector T cells preferentially migrate into the subpial parenchyma from the SAS. Thereafter, suppressor T cells follow, which may lead to the spontaneous recovery from EAE paralysis. PMID- 11419913 TI - Are dopamine receptor agonists neuroprotective in Parkinson's disease? AB - Dopamine receptor agonists are playing an increasingly important role in the treatment of not only patients with advanced Parkinson's disease and those with levodopa-induced motor fluctuations, but also in the early treatment of the disease. This shift has been largely due to the demonstrated levodopa-sparing effect of dopamine agonists and their putative neuroprotective effect, with evidence for the latter being based largely on experimental in vitro and in vivo studies. In this article we review the evidence for neuroprotection by the dopamine agonists pramipexole, ropinirole, pergolide, bromocriptine and apomorphine in cell cultures and animal models of injury to the substantia nigra. Most of the studies suggest that dopamine agonists may have neuroprotective effects via direct scavenging of free radicals or increasing the activities of radical-scavenging enzymes, and enhancing neurotrophic activity. However, the finding that pramipexole can normalise mitochondrial membrane potential and inhibit activity of caspase-3 in cytoplasmic hybrid cells derived from mitochondrial DNA of patients with nonfamilial Alzheimer's disease suggests an even broader implication for the neuroprotective role of dopamine agonists. Although the clinical evidence for neuroprotection by dopamine agonists is still limited, the preliminary results from several ongoing clinical trials are promising. Several longitudinal studies are currently in progress designed to demonstrate a delay or slowing of progression of Parkinson's disease using various surrogate markers of neuronal degeneration such as 18F-levodopa positron emission tomography and 123I beta-CIT (carbomethoxy-beta-4-iodophenyl-nortropane) single positron emission computed tomography. The results of these experimental and clinical studies will improve our understanding of the action of dopamine agonists and provide critical information needed for planning future therapeutic strategies for Parkinson's disease and related neurodegenerative disorders. PMID- 11419914 TI - Dementia with Lewy bodies: clinical features and treatment. AB - Dementia with Lewy bodies (DLB) accounts for 15 to 20% of late-onset dementias. The overlap of cognitive symptoms, neuropsychiatric features, parkinsonism and severe sensitivity to antipsychotic drugs raise a number of key management issues. The neurochemical profile of DLB provides a good theoretical rationale for the potential value of cholinesterase inhibitor therapy, which is supported by clinical evidence from a number of case series and one placebo-controlled double-blind trial. It appears that cholinesterase inhibitor treatment is well tolerated and improves fluctuating confusion, cognition and psychotic symptoms; however, the evidence can still only be considered preliminary and a further double-blind study is imperative. Given the high prevalence of severe sensitivity to antipsychotic drugs in patients with DLB, their role in the treatment of psychiatric symptoms and behavioural problems is uncertain, although a small case report literature indicates that some patients may benefit. On the current balance of evidence, prescription of antipsychotic agents to patients with DLB is not recommended, although further studies focussing on patients with severe and intractable neuropsychiatric symptoms are required. Provisional case series indicate a high degree of motor response to levodopa therapy, although controlled trials are a priority to carefully evaluate the benefits in the context of possible adverse effects, such as the exacerbation of psychosis. PMID- 11419915 TI - Gastro-Oesophageal reflux in the elderly: role of drug therapy in management. AB - Gastro-oesophageal reflux disease (GORD) is common in the elderly. The presenting symptoms of heartburn and regurgitation, so common in the young, are less frequent in the elderly. Common symptoms of GORD in the elderly are dysphagia, vomiting and respiratory problems. Because of the higher risk of associated pathological oesophageal lesions in the older person presenting with symptoms suggestive of GORD, oesophagogastroduodenoscopy must be performed earlier in their clinical course. There is only a poor correlation between the severity of the symptoms and the severity of the associated oesophagitis. Whereas lifestyle modifications are important in individuals with GORD, the use of proton pump inhibitors is recommended to heal the underlying pathology, to resolve the patient's symptoms, to prevent complications, and to improve the quality of life. PMID- 11419916 TI - Optimisation of the prevention and treatment of bacterial endocarditis. AB - This paper reviews currently established guidelines for the prevention and treatment of bacterial endocarditis. Endocarditis remains a life-threatening disease with substantial morbidity and mortality. Primary prevention of endocarditis, whenever possible, is therefore very important. In an individual with endocarditis, rapid diagnosis and effective treatment are essential to good patient outcome. The guidelines discussed here are largely based on those issued by the American Heart Association. While most cases of endocarditis are not attributable to an invasive procedure, certain procedures are associated with bacteraemia by organisms commonly associated with endocarditis, and antibacterial prophylaxis is recommended before such procedures. Patient cardiac conditions are stratified into high, moderate and negligible risk categories based on potential outcome if endocarditis develops. For oral, dental, respiratory tract, and oesophageal procedures (most often associated with viridans streptococci) the standard antibacterial regimen is oral amoxicillin. For gastrointestinal and genitourinary procedures (most often associated with enterococci), parenteral antibacterials are most often recommended. For high-risk patients, intramuscular or intravenous ampicillin and gentamicin (or vancomycin and gentamicin in penicillin-allergic individuals) is recommended. For moderate risk patients, an option of oral amoxicillin or parenteral ampicillin is offered. Treatment of bacterial endocarditis is guided by identification of the causative micro organism. Approximately 80% of cases of endocarditis are due to the gram-positive cocci: streptococci and staphylococci. Other gram-positive organisms include enterococci (predominantly Enterococcusfaecalis and E. faecium) and the HACEK group of organisms (Haemophilus parainfluenzae, H. aphrophilus, Actinobacillus [Haemophilus] actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens, and Kingella kingae). In general, for uncomplicated cases of endocarditis due to penicillin-susceptible viridans streptococci or Streptococcus bovis 4 weeks of benzylpenicillin (or ceftriaxone) is the preferred regimen for most patients aged >65 years. A 2-week course of treatment can be used when gentamicin is added, in patients at low risk for adverse events caused by gentamicin therapy. When endocarditis is caused by strains of viridans streptococci or S. bovis relatively resistant to penicillin, or by enterococci, both benzylpenicillin and gentamicin are recommended. For staphylococcal endocarditis on native heart valves, nafcillin or oxacillin with or without gentamicin is the preferred regimen. In prosthetic valve staphylococcal endocarditis, nafcillin (or oxacillin) with rifampicin and gentamicin is recommended. For all of the above situations, vancomycin is recommended for the patient allergic to penicillin (or methicillin). Finally, consideration of out-of hospital therapy in selected patients is discussed. PMID- 11419917 TI - Endothelin receptor antagonists and cardiovascular diseases of aging. AB - Our understanding of the role of the endothelin system in human cardiovascular physiology and pathophysiology has evolved very rapidly since the initial description of its constituent parts in 1988. Endothelin-1 (ET-1) is the predominant endothelin isoform in the human cardiovascular system and has potent vasoconstrictor, mitogenic and antinatriuretic properties which have implicated it in the pathophysiology of a number of cardiovascular diseases. The effects of ET-1 have been shown to be mediated by 2 principal endothelin receptor subtypes: ET(A) and ET(B). The development of a range of peptidic and nonpeptidic endothelin receptor antagonists represents an exciting breakthrough in human cardiovascular therapeutics. Two main classes of endothelin receptor antagonist have been developed for possible human therapeutic use: ET(A)-selective and nonselective antagonists. Extensive laboratory and clinical research with these agents has highlighted their promise in various cardiovascular diseases. Randomised, placebo-controlled clinical trials have yielded very encouraging results in patients with hypertension and chronic heart failure with more preliminary data suggesting a possible role in the treatment and prevention of atherosclerosis and stroke. Much more research is needed, however, before endothelin receptor antagonists can be considered for clinical use. PMID- 11419918 TI - Inhaled salmeterol: a review of its efficacy in chronic obstructive pulmonary disease. AB - Inhaled salmeterol is a long-acting, selective beta2-adrenoceptor agonist bronchodilator. The drug has been compared with placebo, ipratropium bromide and oral theophylline in patients with chronic obstructive pulmonary disease (COPD) in randomised, clinical trials. Inhaled salmeterol 50 microg twice daily produced significant improvement in forced expiratory volume in 1 second (FEV1), equivalent to that obtained with inhaled ipratropium bromide 40 microg 4 times daily and greater than that obtained with placebo or oral theophylline in randomised trials. Use of as-needed salbutamol (albuterol) was significantly reduced during treatment with inhaled salmeterol or ipratropium bromide compared with placebo or oral theophylline. The time to first COPD exacerbation was significantly longer during 12 weeks of treatment with inhaled salmeterol 50 microg twice daily than ipratropium bromide 40 microg 4 times daily. Compared with baseline and placebo, patients treated for 16 weeks with salmeterol 50 microg (but not 100 microg) twice daily reported significant improvement in total St George's Respiratory Questionnaire (SGRQ) scores. Similarly, more patients treated with inhaled salmeterol 50 microg twice daily or ipratropium bromide 40 microg 4 times daily experienced an increase of > or = 10 points in Chronic Respiratory Disease Questionnaire (CRQ) scores, the minimum clinically significant increment. Compared with placebo, inhaled salmeterol 50 microg twice daily alone, or concurrent with ipratropium bromide 40 microg 4 times daily improved lung function and reduced symptoms in patients with stable COPD in a 12 week, randomised, double-blind study. Clinically meaningful improvement in CRQ scores was documented in significantly more patients treated with the combination of the 2 drugs than either salmeterol monotherapy or placebo. Inhaled salmeterol 50 microg twice daily plus oral theophylline had additive effects on lung function, increased the proportion of symptom-free days and decreased requirements for as-needed salbutamol compared with either agent alone according to a pooled analysis of 2 multicentre, randomised, double-blind studies. CONCLUSION: When used at the optimal dosage, 50 microg twice daily, salmeterol provides symptomatic relief and improves lung function and health-related quality of life in patients with COPD. Available evidence suggests that the drug is as effective as ipratropium bromide and more effective than oral theophylline in patients with COPD. Moreover salmeterol has additive effects when used in combination with inhaled ipratropium bromide or oral theophylline. These qualities make the drug suitable for first-line use in patients with COPD who require regular bronchodilator therapy to manage symptoms. PMID- 11419919 TI - Thyroid carcinoma in single cold nodules and in cold nodules of multinodular goiters. AB - OBJECTIVE: To examine the frequency of malignant growth in a nonfunctioning solitary thyroid nodule (SN) and in multinodular goiter (MNG). METHODS: We performed fine-needle aspiration biopsy (FNAB) of cold nodules in 154 patients. Eight patients with unsatisfactory FNAB specimens were excluded from the study. Of the remaining 146 patients, 95 (89 women and 6 men) had MNG. The other 51 patients (46 women and 5 men) had SN. The overall age range was 25 to 86 years (mean, 52). RESULTS: Of the 95 patients with MNG, 16 had abnormal cytologic findings and underwent surgical excision. Of these 16 patients, 12 had thyroid cancer (6 papillary, 3 follicular, and 3 Hurthle cell). Of the 51 patients with SN, 8 had suspicious cytologic findings, and 5 had surgical histologic evidence of thyroid cancer (3 papillary and 2 follicular). Four patients with papillary thyroid cancer (three from the MNG group and one from the SN group) had a history of exposure to neck irradiation. After exclusion of the patients with a history of neck irradiation, the rate of malignant involvement in cold nodules in the MNG group was 9.78%, in comparison with 8% in the group with SN (P = 0.89). Within the MNG group, 25% of the thyroid malignant lesions were of the Hurthle cell type. CONCLUSION: Multinodularity of a goiter should no longer be considered an indicator of probable benign disease. In the assessment of all thyroid nodules, both SN and those in MNG, a thyroid scan can be helpful, and it should be followed by FNAB and cytopathologic examination of any nonfunctioning lesions. The incidence of malignant involvement in cold nodules of MNG does not differ significantly from that found in SN. PMID- 11419920 TI - Pitfalls of conventional human chorionic gonadotropin stimulation test to detect hormonally functional cryptorchid testes in midchildhood. AB - OBJECTIVE: To report two cases misdiagnosed as bilateral anorchism in midchildhood on the basis of multiple conventional human chorionic gonadotropin (HCG) stimulation tests and sonograms of the abdomen and pelvis. METHODS: In two young male patients with cryptorchidism who were considered to have anorchism, we describe the findings on clinical examination, the testosterone levels before and after standard HCG stimulation testing, and sonographic findings during the midchildhood period. In both cases, as the children approached puberty the diagnosis was found to be incorrect. RESULTS: Two boys, 8 and 91/2 years old, were seen in consultation in our Pediatric Endocrine Clinic with a presumed diagnosis of anorchism. In the first case, multiple conventional HCG stimulation tests were done. In the second case, a single stimulation test was performed during routine follow-up assessments. In both cases, testosterone levels before and after HCG stimulation were consistent with the diagnosis of absent functional testicular tissue. Sonograms of the abdomen and pelvis also failed to detect the testicles. Both patients were ultimately noted to be pubertal (at 14 1/2 and >13 1/2 years, respectively) and to have early pubertal testosterone levels. A testicle was detected in one patient by abdominal computed tomographic scan and in the other by palpation of the inguinal canal. CONCLUSION: Conventional dosing and duration of the HCG stimulation test, as widely recommended in standard textbooks and in articles in the medical literature, may not elicit positive HCG induced testosterone responses during midchildhood for detection of functional testicular tissue. During the midchildhood period, which is characterized by low gonadotropin, low sex steroid production, and a highly sensitive hypothalamic pituitary-gonadal axis to feedback inhibition, a prolonged HCG stimulation test perhaps of 4 to 6 weeks' duration-may be necessary. In addition, other investigational modalities may need to be used to detect the presence of functional testicular tissue during this developmental period. PMID- 11419921 TI - Genetic, clinical, and biochemical analysis of unrelated Spanish families with multiple endocrine neoplasia type I. AB - OBJECTIVE: (1) To study seven unrelated Spanish families with multiple endocrine neoplasia type I (MEN I), describing clinical features and investigating the presence of germline mutations in the MEN1 gene, and (2) to establish reference values for pancreatic polypeptide and gastrin after a standardized test meal in a healthy control group, analyzing the usefulness of this test for detecting neuroendocrine gastroenteropancreatic tumors in subjects with MEN I. METHODS: Two or three generations of 7 kindreds with MEN I, consisting of a total of 39 individual family members, were investigated. Three of the families were subjected only to genetic analysis, and the other four families were also assessed clinically. A group of 23 healthy control subjects were also studied. RESULTS: Mutations in the MEN1 gene were found in six of the seven families studied. Of the 4 families studied clinically, 12 family members were genetically affected. In these study subjects, hyperparathyroidism, adrenal adenomas, neuroendocrine gastroenteropancreatic tumors, and pituitary adenomas developed in 100%, 50%, 16%, and 12%, respectively. All demonstrated pancreatic tumors were associated with abnormal results after a test meal, but 75% of them also showed high basal hormonal measurements. CONCLUSION: Analysis of the MEN1 gene decreases the total number of subjects who need to undergo repeated clinical and biochemical studies, but genetic mutations are not detected in all families with MEN I. Hyperparathyroidism is the most common manifestation of the syndrome, but the presence of adrenal adenomas has probably been underestimated. Ingestion of a standardized test meal for stimulation of gastrin and pancreatic polypeptide could be a complementary procedure for diagnosing gastroenteropancreatic tumors in selected patients with MEN I in whom basal gastrin and pancreatic polypeptide levels are normal. PMID- 11419922 TI - Primary, secondary, tertiary, and quaternary treatment with troglitazone in type 2 diabetes mellitus in an outpatient clinical practice. AB - OBJECTIVE: To assess the value of troglitazone in various combinations of therapy with sulfonylureas, biguanides, and insulin, including triple oral agent therapy with and without insulin, in an outpatient clinical practice. METHODS: We compiled results in our first 207 patients with type 2 diabetes treated for 1 year with troglitazone. In eight study groups, troglitazone was added to the following prior regimens: (1) diet alone, (2) sulfonylurea, (3) metformin, (4) sulfonylurea and metformin, (5) insulin, (6) insulin and sulfonylurea, (7) insulin and metformin, and (8) insulin, sulfonylurea, and metformin. Glycosylated hemoglobin, fasting plasma glucose, hemoglobin, weight, and insulin dose were recorded every 3 months for 1 year and are reported. RESULTS: The glycosylated hemoglobin decreased in all study groups during the 1-year period. With the addition of troglitazone, it declined from 8.0% to 6.0% in the diet-only treatment group, from 8.9% to 7.3% in the sulfonylurea group, from 8.4% to 7.2% in the metformin group, and from 8.6% to 7.5% in the group treated with sulfonylurea and metformin. For the groups receiving insulin, glycosylated hemoglobin decreased in conjunction with a significant decline in insulin dose. The dose of insulin was reduced from a baseline of 62 to 41 U per patient in the insulin-only treatment group, from 36 to 12 U per patient in the group treated with insulin and sulfonylurea, and from 28 to 11 U per patient in the group treated with insulin and metformin. In the group treated with insulin, sulfonylurea, and metformin, the dose of insulin was decreased from 36 to 13 U. CONCLUSION: From our data in more than 200 patients with type 2 diabetes, who participated in a 1-year follow-up in an outpatient clinical practice, we found that troglitazone improved glycemic control in each study group and also decreased the insulin dose in the insulin-requiring patients. Troglitazone improves glycemic control as a primary, secondary, tertiary, or even quaternary therapy in patients with type 2 diabetes. PMID- 11419923 TI - Central diabetes insipidus due to herpes simplex in a patient immunosuppressed by Cushing's syndrome. AB - OBJECTIVE: To describe a patient immunocompromised by Cushing's syndrome in whom central diabetes insipidus developed as a complication of herpes simplex involvement of the hypothalamus. METHODS: We present a case, including results of laboratory and histopathologic studies, in which herpes simplex was established as the causative agent for central diabetes insipidus. RESULTS: A woman with ectopic corticotropin-dependent Cushing's syndrome, diabetes mellitus, carcinoid tumor, and a history of thyroid cancer had the precipitous onset of seizure and fever, and hypotonic polyuria and progressive hypernatremia developed. Central diabetes insipidus was diagnosed and successfully treated with desmopressin. Nevertheless, the patient's condition deteriorated and she died. Autopsy revealed herpes simplex encephalitis involving the magnicellular neurons of the hypothalamus. CONCLUSION: Central diabetes insipidus caused by viral infections has been reported in immunosuppressed patients, such as those with acquired immunodeficiency syndrome (AIDS). To our knowledge, this is the first report of a herpes infection causing diabetes insipidus in a patient immunosuppressed by Cushing's syndrome. This case demonstrates that, in patients with Cushing's syndrome, diabetes insipidus may develop as a result of herpes simplex infection of the hypothalamus. PMID- 11419924 TI - An unusual case of inappropriate secretion of thyrotropin: neoplastic or nonneoplastic? AB - OBJECTIVE: To report an unusual case that illustrates the difficulties in distinguishing neoplastic (nIST) from nonneoplastic inappropriate secretion of thyrotropin (nnIST). METHODS: We describe clinical, biochemical, genetic analysis, and magnetic resonance imaging (MRI) results in a patient with hyperthyroidism due to IST, and we review the related literature. RESULTS: The patient demonstrated overt clinical and biochemical hyperthyroidism with inappropriately increased thyrotropin levels, which failed to respond to thyrotropin-releasing hormone (TRH) stimulation. Sex hormone-binding globulin (SHBG) levels were in the hyperthyroid range. Alpha subunit levels were normal, as was the alpha subunit/thyrotropin molar ratio. MRI of the pituitary was negative for tumor during a 2-year period, and octreotide scan was also negative for sellar uptake. Basal oxygen consumption was abnormally increased. Genetic analysis failed to reveal mutations of the thyroid receptor b gene. The patient responded well to radioiodine ablation of his thyroid. CONCLUSION: This patient had clinical symptoms of hyperthyroidism associated with some features characteristic of nIST (increased level of SHBG, lack of thyrotropin response to TRH stimulation, absence of thyroid receptor b mutations) and others typical of nnIST (normal alpha subunit and its molar ratio to thyrotropin, absence of tumor on sellar imaging). Close follow-up with periodic MRI of the sella is important because of the possible existence of a small pituitary tumor, which may become apparent at a later date. Therapy to control symptoms is important. Hormone replacement, if needed, should be adjusted to maintain clinical euthyroidism, guided by free thyroxine levels. PMID- 11419925 TI - Successful outcome of pregnancy in a patient with generalized lipoatrophic diabetes mellitus. AB - OBJECTIVE: To report the first known case of a successful outcome of pregnancy in a patient with generalized lipoatrophic diabetes. METHODS: We present a detailed case report of a patient who achieved and successfully completed a pregnancy despite having lipoatrophic diabetes. RESULTS: With careful attention to glycemic control with use of U-500 insulin and strict avoidance of dietary fat, a 23-year old woman with lipoatrophic diabetes maintained a pregnancy to 28 weeks. The infant weighed 1,235 g and was devoid of serious metabolic complications. Three months after childbirth, the patient died of gastrointestinal bleeding. CONCLUSION: We are not aware of any previously published report of a successful pregnancy in a patient with generalized, acquired lipoatrophic diabetes. Because of the involvement of multiple organ systems in generalized lipoatrophic diabetes, female patients should be thoroughly advised of the serious pregnancy associated risks to both the mother and the fetus and the need for extremely close monitoring. PMID- 11419926 TI - Radioiodine uptake in the head and neck. AB - OBJECTIVE: To report two cases of sinusitis-associated radioiodine uptake in patients with thyroid cancer and to review the reported causes of false-positive radioiodine uptake in the head and neck area. METHODS: We present the radiologic findings in two patients who had undergone treatment for papillary thyroid cancer and discuss other settings in which radioiodine uptake suggested the presence of metastatic disease. RESULTS: Radioiodine whole-body scans of two patients who had had thyroid cancer demonstrated uptake in the sphenoid and maxillary sinuses, respectively, mimicking bone or brain metastatic involvement. The thyroglobulin levels were low. Computed tomographic (CT) scanning disclosed mucosal swelling in the sinuses, consistent with sinusitis. The radioiodine uptake cleared on a follow-up scan in one case and was more localized than the CT findings in the other. Eighteen causes of false-positive radioiodine uptake in the head and neck area have been reported. On the basis of the mechanism of uptake, they can be classified into four categories: (1) physiologic uptake (ectopic thyroid tissue), (2) nonthyroidal pathologic conditions (dacryocystitis, sinusitis, sinus mucocele, sialadenitis, folliculitis, Warthin's tumor, parotid cyst, porencephaly, posttraumatic cerebromalacia, and inflammation due to dental disease or a nose ring), (3) internal retention (ectasia of the carotid artery and an artificial eye), and (4) external contamination by body secretions (sweat and nasal, tracheobronchial, lacrimal, and salivary secretions). The estimated prevalence of external contamination in the head and neck area on whole-body radioiodine scans is 0.3%. CONCLUSION: Physicians should rule out the presence of radioiodine uptake by inflamed mucosa of the paranasal sinuses, as well as various other causes of false-positive radioiodine uptake, before metastatic thyroid cancer in the head and neck area is diagnosed. PMID- 11419927 TI - Basal insulin replacement and use of rapid-acting insulin analogues in patients with type 1 diabetes. AB - OBJECTIVE: To analyze various treatment strategies for replacement of basal insulin in patients with type 1 diabetes who are receiving rapid-acting insulin with meals. METHODS: Recently published studies are reviewed, and the available information on basal insulin adjustment is summarized. RESULTS: In a comparison of presupper insulin lispro versus regular human insulin in adolescent patients, postprandial blood glucose excursion was much lower with insulin lispro. Patients who were given regular insulin at supper and no bedtime snack had a significantly higher occurrence of nocturnal hypoglycemia than did those given insulin lispro at supper and no snack at bedtime. In another study that attempted to identify the optimal basal insulin in the context of multidose therapy with insulin lispro, NPH insulin yielded lower blood glucose levels and a trend toward better glycosylated hemoglobin levels than did Ultralente insulin. For improvement of glycemic control during insulin lispro therapy, adjustments in the ratio of bolus to basal insulin and the number of basal insulin injections have been shown to be useful. In addition, use of insulin lispro in continuous subcutaneous insulin infusion therapy significantly reduced the rate of occurrence of hypoglycemic episodes in comparison with regular insulin. CONCLUSION: Rapid-acting insulin analogues improve postprandial blood glucose control and decrease the frequency of hypoglycemia in patients with type 1 diabetes whether insulin pump therapy or a multidose insulin injection regimen is being used. For full benefit of short acting insulin analogues, adjustments must be made in basal insulin replacement. PMID- 11419928 TI - Optimization of insulin therapy in patients with type 2 diabetes. AB - OBJECTIVE: To analyze emerging trends in effective utilization of insulin in the treatment of type 2 diabetes. METHODS: A systematic management plan and requirements for successful insulin therapy in patients with type 2 diabetes are outlined. RESULTS: The key to successful management of type 2 diabetes with insulin therapy is the use of adequate amounts of insulin and a bolus-to-basal insulin ratio of 1:1. Patient education about the progressive nature of the disease and the importance of self-monitoring of blood glucose is essential. The most effective method of sustaining normal blood glucose levels is to mimic the physiologic response to intake of food and thereby control postprandial glycemic excursions. Rapid-acting insulin lispro is useful in this effort. In the development of effective insulin regimens, one option is use of an insulin sensitizer in combination with bedtime NPH insulin. CONCLUSION: Because of deteriorating beta cell function, insulin deficiency results and usually necessitates administration of exogenous insulin in patients with type 2 diabetes. The goal of insulin treatment of type 2 diabetes is to achieve near normal glycemic control. The insulin secretory defect progresses over time; thus, therapy must be continually matched to the underlying pathophysiologic defects. PMID- 11419929 TI - Optimization of insulin therapy in patients with gestational diabetes. AB - OBJECTIVE: To discuss the new criteria for screening and diagnosis of gestational diabetes and present recommendations for patient management. METHODS: Various strategies for detecting gestational diabetes historically and currently are reviewed, and approaches for optimizing patient and neonatal outcome are discussed. RESULTS: Rather than testing for gestational diabetes selectively, universal screening of pregnant women between 24 and 28 weeks of gestation with use of a glucose challenge test and 1-hour assessment has been shown to decrease the rate of occurrence of neonatal macrosomia. Performance of both preprandial and postprandial self-monitoring of blood glucose levels helps to prevent pronounced postprandial glucose excursions and minimize the risk of neonatal macrosomia. In comparison with regular human insulin, insulin lispro has yielded significantly better metabolic control and a reduction in hypoglycemic episodes before breakfast in patients with gestational diabetes. CONCLUSION: Women with gestational diabetes who are not optimally managed with diet and exercise need insulin therapy. Insulin lispro is associated with fewer hypoglycemic events and attenuates the postprandial response greater than does regular human insulin. PMID- 11419930 TI - Consumer-driven changes in health care in the United States. PMID- 11419931 TI - Is thyroid carcinoma more common in single nodules or multinodular goiters? PMID- 11419932 TI - Appropriate use and interpretation of human chorionic gonadotropin stimulation in prepubertal male patients. PMID- 11419933 TI - The RNA polymerase III transcription apparatus. PMID- 11419934 TI - Comparison between long-range interactions and contact order in determining the folding rate of two-state proteins: application of long-range order to folding rate prediction. AB - The contact order is believed to be an important factor for understanding protein folding mechanisms. In our earlier work, we have shown that the long-range interactions play a vital role in protein folding. In this work, we analyzed the contribution of long-range contacts to determine the folding rate of two-state proteins. We found that the residues that are close in space and are separated by at least ten to 15 residues in sequence are important determinants of folding rates, suggesting the presence of a folding nucleus at an interval of approximately 25 residues. A novel parameter "long-range order" has been proposed to predict protein folding rates. This parameter shows as good a relationship with the folding rate of two-state proteins as contact order. Further, we examined the minimum limit of residue separation to determine the long-range contacts for different structural classes. We observed an excellent correlation between long-range order and folding rate for all classes of globular proteins. We suggest that in mixed-class proteins, a larger number of residues can serve as folding nuclei compared to all-alpha and all-beta proteins. A simple statistical method has been developed to predict the folding rates of two-state proteins using the long-range order that produces an agreement with experimental results that is better or comparable to other methods in the literature. PMID- 11419935 TI - Interactions of an Arg-rich region of transcription elongation protein NusA with NUT RNA: implications for the order of assembly of the lambda N antitermination complex in vivo. AB - The E. coli NusA transcription elongation protein (NusA(Ec)), identified because of its requirement for transcription antitermination by the N protein, has an Arg rich S1 RNA-binding domain. A complex of N and NusA with other host factors binding at NUT sites in the RNA renders RNA polymerase termination-resistant. An E. coli haploid for nusA944, having nine different codons replacing four normally found in the Arg-rich region, is defective in support of N action. Another variant, haploid for the nusAR199A allele, with a change in a highly conserved Arg codon in the S1 domain, effectively supports N-mediated antitermination. However, nusAR199A is recessive to nusA944, while nusA(Ec) is dominant to nusA944 for support of N-mediated antitermination, suggesting a competition between NusA944 and NusAR199A during complex formation. Complex formation with the variant NusA proteins was assessed by mobility gel shifts. NusAR199A, unlike NusA(Ec) and NusA944, fails to form a complex with N and NUT RNA. However, while NusAR199A, like wild-type NusA, forms an enlarged complex with NUT RNA, N, RNA polymerase, and other host proteins required for efficient N-mediated antitermination, NusA944 does not form this enlarged complex. Consistent with the in vivo results, NusA944 prevents NusAR199A but not NusA(Ec) from forming the enlarged complex. The simplest conclusion from these dominance studies is that in the formation of the complete active antitermination complex in vivo, NusA and N binding to the newly synthesized NUT RNA precedes addition of the other factors. Alternative less effective routes to the active complex that allows bypass of this preferred pathway may also exist. PMID- 11419936 TI - A small protein-protein interaction domain common to KlcB and global regulators KorA and TrbA of promiscuous IncP plasmids. AB - The kor regulon of broad host-range, incompatibility group P (IncP) plasmids uses the KorA, KorB, and KorC repressors to regulate expression of genes for replication, conjugation, segregation, and host range. One operon, kilC, encodes the KorC repressor and two genes of unknown function (klcA and klcB). The predicted sequences of the 51.1 kDa KlcB protein, the 11.3 kDa KorA repressor, and another small (13.5 kDa) regulatory protein, TrbA, show a highly related 35 amino acid residue segment (V-L-P domain). We found that induction of the klcB gene is toxic to Escherichia coli host cells harboring an IncP plasmid. We confirmed a model in which the V-L-P domain of KlcB interacts directly with the V L-P domain of KorA to derepress KorA-regulated operons, thereby allowing expression of toxic genes. First, a lacZ reporter fused to the kleA promoter, which is regulated by KorA and KorC, revealed that klcB induction specifically releases KorA-repression but has no effect on KorC repression. Second, induced expression of the V-L-P domains from KorA or KlcB is sufficient to release KorA repression at the kleA promoter. Third, purified GST-KlcB fusion protein interacts specifically with His-tagged KorA. Fourth, fusion of the V-L-P domains of KorA and TrbA and full-length KlcB polypeptide to the DNA-binding domain of bacteriophage lambda repressor leads to the formation of functional, dimeric repressors, and mutations that alter conserved residues of the V-L-P domain adversely affect dimerization. Fifth, crosslinking experiments demonstrated that the V-L-P domain of KorA is able to dimerize in solution and form heterodimers in mixtures with full-length KorA polypeptide. These findings show that the V-L-P domain is a protein-protein interaction module that is likely to be responsible for dimerization of KorA and TrbA, and important for KlcB dimerization. We speculate on the possible significance of KlcB-KorA heterodimers in IncP plasmid maintenance. PMID- 11419937 TI - DnaA boxes in the P1 plasmid origin: the effect of their position on the directionality of replication and plasmid copy number. AB - The DnaA protein is essential for initiation of DNA replication in a wide variety of bacterial and plasmid replicons. The replication origin in these replicons invariably contains specific binding sites for the protein, called DnaA boxes. Plasmid P1 contains a set of DnaA boxes at each end of its origin but can function with either one of the sets. Here we report that the location of origin opening, initiation site of replication forks and directionality of replication do not change whether the boxes are present at both or at one of the ends of the origin. Replication was bidirectional in all cases. These results imply that DnaA functions similarly from the two ends of the origin. However, origins with DnaA boxes proximal to the origin-opening location opened more efficiently and maintained plasmids at higher copy numbers. Origins with the distal set were inactive unless the adjacent P1 DNA sequences beyond the boxes were included. At either end, phasing of the boxes with respect to the remainder of the origin influenced the copy number. Thus, although the boxes can be at either end, their precise context is critical for efficient origin function. PMID- 11419938 TI - Conformational changes of the ferric uptake regulation protein upon metal activation and DNA binding; first evidence of structural homologies with the diphtheria toxin repressor. AB - Fur (ferric uptake regulation protein) is a bacterial global regulator that uses iron as a cofactor to bind to specific DNA sequences. It has been suggested that metal binding induces a conformational change in the protein, which is subsequently able to recognize DNA. This mechanism of activation has been investigated here using selective chemical modification monitored by mass spectrometry. The reactivity of each lysine residue of the Fur protein was studied, first in the apo form of the protein, then after metal activation and finally after DNA binding. Of particular interest is Lys76, which was shown to be highly protected from modification in the presence of target DNA. Hydrogen deuterium exchange experiments were performed to map with higher resolution the conformational changes induced by metal binding. On the basis of these results, together with a secondary structure prediction, the presence in Fur of a non classical helix-turn-helix motif is proposed. Experimental results show that activation upon metal binding induces conformational modification of this specific motif. The recognition helix, interacting directly with the major groove of the DNA, would include the domain [Y55-F61]. This helix would be followed by a small "wing" formed between two beta strands, containing Lys76, which might interact directly with DNA. These results suggest that Fur and DtxR (diphtheria toxin repressor), another bacterial repressor, share not only the function of being iron concentration regulators, and the structure of their DNA-binding domain. PMID- 11419939 TI - S-adenosyl-L-methionine is required for DNA cleavage by type III restriction enzymes. AB - The requirement of S-adenosyl-L-methionine (AdoMet) in the cleavage reaction carried out by type III restriction-modification enzymes has been investigated. We show that DNA restriction by EcoPI restriction enzyme does not take place in the absence of exogenously added AdoMet. Interestingly, the closely related EcoP15I enzyme has endogenously bound AdoMet and therefore does not require the addition of the cofactor for DNA cleavage. By employing a variety of AdoMet analogs, which differ structurally from AdoMet, this study demonstrates that the carboxyl group and any substitution at the epsilon carbon of methionine is absolutely essential for DNA cleavage. Such analogs could bring about the necessary conformational change(s) in the enzyme, which make the enzyme proficient in DNA cleavage. Our studies, which include native polyacrylamide gel electrophoresis, molecular size exclusion chromatography, UV, fluorescence and circular dichroism spectroscopy, clearly demonstrate that the holoenzyme and apoenzyme forms of EcoP15I restriction enzyme have different conformations. Furthermore, the Res and Mod subunits of the EcoP15I restriction enzyme can be separated by gel filtration chromatography in the presence of 2 M NaCl. Reconstitution experiments, which involve mixing of the isolated subunits, result in an apoenzyme form, which is restriction proficient in the presence of AdoMet. However, mixing the Res subunit with Mod subunit deficient in AdoMet binding does not result in a functional restriction enzyme. These observations are consistent with the fact that AdoMet is required for DNA cleavage. In vivo complementation of the defective mod allele with a wild-type mod allele showed that an active restriction enzyme could be formed. Furthermore, we show that while the purified c2-134 mutant restriction enzyme is unable to cleave DNA, the c2-440 mutant enzyme is able to cleave DNA albeit poorly. Taken together, these results suggest that AdoMet binding causes conformational changes in the restriction enzyme and is necessary to bring about DNA cleavage. PMID- 11419940 TI - Derivation of a structural model for the c-myc IRES. AB - We have derived a secondary structure model for the c-myc internal ribosome entry segment (IRES) by using information from chemical probing of the c-myc IRES RNA to constrain structure prediction programs. Our data suggest that the IRES is modular in nature, and can be divided into two structural domains linked by a long unstructured region. Both domains are required for full IRES function. Domain 1 is a complex element that contains a GNNRA apical loop and an overlapping double pseudoknot motif that is topologically unique amongst published RNA structures. Domain 2, the smaller of the two, contains an apical AUUU loop. We have located the ribosome landing site and have shown that ribosomes enter in a 16 nt region downstream of the pseudoknots in a situation similar to that observed in several viral IRESs. To test the structure, several key regions of the IRES were mutated and, interestingly, it appears that some of the structural elements that we have identified function to repress c-myc IRES function. This has profound implications for de-regulation of c-myc expression by mutations occurring in the IRES. PMID- 11419941 TI - cDNA and genomic cloning of lacritin, a novel secretion enhancing factor from the human lacrimal gland. AB - Multiple extracellular factors are hypothesized to promote the differentiation of unstimulated and/or stimulated secretory pathways in exocrine secretory cells, but the identity of differentiation factors, particularly those organ-specific, remain largely unknown. Here, we report on the identification of a novel secreted glycoprotein, lacritin, that enhances exocrine secretion in overnight cultures of lacrimal acinar cells which otherwise display loss of secretory function. Lacritin mRNA and protein are highly expressed in human lacrimal gland, moderately in major and minor salivary glands and slightly in thyroid. No lacritin message or protein is detected elsewhere among more than 50 human tissues examined. Lacritin displays partial similarity to the glycosaminoglycan binding region of brain-specific neuroglycan C (32 % identity over 102 amino acid residues) and to the possibly mucin-like amino globular region of fibulin-2 (30 % identity over 81 amino acid residues), and localizes primarily to secretory granules and secretory fluid. The lacritin gene consists of five exons, displays no alternative splicing and maps to 12q13. Recombinant lacritin augments unstimulated but not stimulated acinar cell secretion, promotes ductal cell proliferation, and stimulates signaling through tyrosine phosphorylation and release of calcium. It binds collagen IV, laminin-1, entactin/nidogen-1, fibronectin and vitronectin, but not collagen I, heparin or EGF. As an autocrine/paracrine enhancer of the lacrimal constitutive secretory pathway, ductal cell mitogen and stimulator of corneal epithelial cells, lacritin may play a key role in the function of the lacrimal gland-corneal axis. PMID- 11419942 TI - Vps9, Rabex-5 and DSS4: proteins with weak but distinct nucleotide-exchange activities for Rab proteins. AB - The activities of three Rab-specific factors with GDP/GTP exchange activity, Vps9p, Rabex-5 and DSS4, with their cognate GTPases, Ypt51p, Rab5 and Ypt1p, have been analysed quantitatively. In contrast to other exchange factors examined and to DSS4, Vps9p, and by analogy probably Rabex-5, have considerably lower affinity than GDP to the respective GTPases. In keeping with this, they are relatively weak exchangers, with a maximal rate constant for GDP release from the ternary complex between exchange factor, GTPase and GDP of ca 0.01 s(-1), which is several orders of magnitude lower than for other exchange factors examined. If interaction with these proteins is a mandatory aspect of the Rab cycle, this suggests that the overall rate of cycling might be controlled at this point of the cycle. Surprisingly, DSS4, which has the thermodynamic potential to displace GDP effectively from Ypt1p, also does this very slowly, again with a maximal rate constant of ca 0.01 s(-1). An additional, and based on present knowledge, unique, feature of the Ypt1p.DSS4 complex, is that the association of GTP (or GDP) is more than 10(3)-fold slower than to Ypt1p, thus leading to a long life-time of the binary complex between the two proteins, even at the high nucleotide concentrations that prevail in the cell. This leads to the conclusion that the protein-protein complex is likely to have an important biological significance in addition to its probable role in GTP/GDP exchange. PMID- 11419943 TI - Non-fibrillar oligomeric species of the amyloid ABri peptide, implicated in familial British dementia, are more potent at inducing apoptotic cell death than protofibrils or mature fibrils. AB - Familial British dementia (FBD) is an autosomal dominant neurodegenerative disorder, with biochemical and pathological similarities to Alzheimer's disease. FBD is associated with a point mutation in the stop codon of the BRI gene. The mutation extends the length of the wild-type protein by 11 amino acids, and following proteolytic cleavage, results in the production of a cyclic peptide (ABri) 11 amino acids longer than the wild-type (WT) peptide produced from the normal gene BRI. ABri was found to be the main component of amyloid deposits in FBD brains. However, pathological examination of FBD brains has shown the presence of ABri as non-fibrillar deposits as well as amyloid fibrils. Taken together, the genetic, pathological and biochemical data support the hypothesis that ABri deposits play a central role in the pathogenesis of FBD. Here we report that ABri, but not WT peptide, can oligomerise and form amyloid-like fibrils. We show for the first time that ABri induces apoptotic cell death, whereas WT is not toxic to cells. Moreover, we report the novel findings that non-fibrillar oligomeric species of ABri are more toxic than protofibrils and mature fibrils. These findings provide evidence that non-fibrillar oligomeric species are likely to play a critical role in the pathogenesis of FBD and suggest that a similar process may also operate in other neurodegenerative diseases. PMID- 11419944 TI - Modulation of antigenicity related to changes in antibody flexibility upon lyophilization. AB - Lyophilization is frequently used to increase the shelf-life of biopharmaceuticals containing antibodies. A case in which an anti-idiotypic antibody, MMA 383, substantially lost its in vivo immunogenic properties although the protein was not degraded, is investigated. The scanning transmission electron microscope allowed the MMA 383 Fab and Fc moieties to be resolved. By averaging the single antibodies, the angle between the Fab moieties can be calculated. Non lyophilized antibodies displayed a wider range of shapes than their reconstituted, lyophilized counterparts. Accordingly, the angle between the two Fab fragments varied more, indicating greater flexibility. The tryptophan steady state fluorescence intensity, steady-state fluorescence anisotropy and fluorescence lifetime, were smaller for the lyophilized antibodies. These were also more resistant towards thermal denaturation/aggregation. Circular dichroism spectra detected temperature-dependent differences between the two antibody types in the 236 nm region. The subtle but reproducible structural changes induced by lyophilization may be related to the loss of in vivo immunogenic properties. PMID- 11419945 TI - A double chain reversal loop and two diagonal loops define the architecture of a unimolecular DNA quadruplex containing a pair of stacked G(syn)-G(syn)-G(anti) G(anti) tetrads flanked by a G-(T-T) Triad and a T-T-T triple. AB - The architecture of G-G-G-G tetrad-aligned DNA quadruplexes in monovalent cation solution is dependent on the directionality of the four strands, which in turn are defined by loop connectivities and the guanine syn/anti distribution along individual strands and within individual G-G-G-G tetrads. The smallest unimolecular G-quadruplex belongs to the d(G2NnG2NnG2NnG2) family, which has the potential to form two stacked G-tetrads linked by Nn loop connectivities. Previous studies have focused on the thrombin-binding DNA aptamer d(G2T2G2TGTG2T2G2), where Nn was T2 for the first and third connecting loops and TGT for the middle connecting loop. This DNA aptamer in K(+) cation solution forms a unimolecular G-quadruplex stabilized by two stacked G(syn)-G(anti)-G(syn) G(anti) tetrads, adjacent strands which are antiparallel to each other and edge wise connecting T2, TGT and T2 loops. We now report on the NMR-based solution structure of the d(G2T4G2CAG2GT4G2T) sequence, which differs from the thrombin binding DNA aptamer sequence in having longer first (T4) and third (GT4) loops and a shorter (CA) middle loop. This d(G2T4G2CAG2GT4G2T) sequence in Na(+) cation solution forms a unimolecular G-quadruplex stabilized by two stacked G(syn) G(syn)-G(anti)-G(anti) tetrads, adjacent strands which have one parallel and one antiparallel neighbors and distinct non-edge-wise loop connectivities. Specifically, the longer first (T4) and third (GT4) loops are of the diagonal type while the shorter middle loop is of the double chain reversal type. In addition, the pair of stacked G-G-G-G tetrads are flanked on one side by a G-(T T) triad and on the other side by a T-T-T triple. The distinct differences in strand directionalities, loop connectivities and syn/anti distribution within G-G G-G tetrads between the thrombin-binding DNA aptamer d(G2T2G2TGTG2T2G2) quadruplex reported previously, and the d(G2T4G2CAG2GT4G2T) quadruplex reported here, reinforces the polymorphic nature of higher-order DNA architectures. Further, these two small unimolecular G-quadruplexes, which are distinct from each other and from parallel-stranded G-quadruplexes, provide novel targets for ligand recognition. Our results demonstrate that the double chain reversal loop connectivity identified previously by our laboratory within the Tetrahymena telomere d(T2G4)4 quadruplex, is a robust folding topology, since it has now also been observed within the d(G2T4G2CAG2GT4G2T) quadruplex. The identification of a G-(T-T) triad and a T-T-T triple, expands on the available recognition alignments for base triads and triples. PMID- 11419946 TI - Crystal structure of thiamin pyrophosphokinase. AB - Thiamin pyrophosphate (TPP) is a coenzyme derived from vitamin B1 (thiamin). TPP synthesis in eukaryotes requires thiamin pyrophosphokinase (TPK), which catalyzes the transfer of a pyrophosphate group from ATP to thiamin. TPP is essential for central metabolic processes, including the formation of acetyl CoA from glucose and the Krebs cycle. Deficiencies in human thiamin metabolism result in beriberi and Wernicke encephalopathy. The crystal structure of mouse TPK was determined by multiwavelength anomalous diffraction at 2.4 A resolution, and the structure of TPK complexed with thiamin has been refined at 1.9 A resolution. The TPK polypeptide folds as an alpha/beta-domain and a beta-sandwich domain, which share a central ten-stranded mixed beta-sheet. TPK subunits associate as a dimer, and thiamin is bound in the dimer interface. Despite lacking apparent sequence homology with other proteins, the alpha/beta-domain resembles the Rossman fold and is similar to other kinase structures, including another pyrophosphokinase and a thiamin biosynthetic enzyme. Comparison of mouse and yeast TPK structures reveals differences that could be exploited in developing species-specific inhibitors of potential use as antimicrobial agents. PMID- 11419947 TI - Crystal structure of lactose synthase reveals a large conformational change in its catalytic component, the beta1,4-galactosyltransferase-I. AB - The lactose synthase (LS) enzyme is a 1:1 complex of a catalytic component, beta1,4-galactosyltransferse (beta4Gal-T1) and a regulatory component, alpha lactalbumin (LA), a mammary gland-specific protein. LA promotes the binding of glucose (Glc) to beta4Gal-T1, thereby altering its sugar acceptor specificity from N-acetylglucosamine (GlcNAc) to glucose, which enables LS to synthesize lactose, the major carbohydrate component of milk. The crystal structures of LS bound with various substrates were solved at 2 A resolution. These structures reveal that upon substrate binding to beta4Gal-T1, a large conformational change occurs in the region comprising residues 345 to 365. This repositions His347 in such a way that it can participate in the coordination of a metal ion, and creates a sugar and LA-binding site. At the sugar-acceptor binding site, a hydrophobic N-acetyl group-binding pocket is found, formed by residues Arg359, Phe360 and Ile363. In the Glc-bound structure, this hydrophobic pocket is absent. For the binding of Glc to LS, a reorientation of the Arg359 side-chain occurs, which blocks the hydrophobic pocket and maximizes the interactions with the Glc molecule. Thus, the role of LA is to hold Glc by hydrogen bonding with the O-1 hydroxyl group in the acceptor-binding site on beta4Gal-T1, while the N-acetyl group-binding pocket in beta4Gal-T1 adjusts to maximize the interactions with the Glc molecule. This study provides details of a structural basis for the partially ordered kinetic mechanism proposed for lactose synthase. PMID- 11419948 TI - Solution structure of human IL-13 and implication for receptor binding. AB - Interleukin-13 has been implicated as a key factor in asthma, allergy, atopy and inflammatory response, establishing the protein as a valuable therapeutic target. The high-resolution solution structure of human IL-13 has been determined by multidimensional NMR. The resulting structure is consistent with previous short chain left-handed four-helix bundles, where a significant similarity in the folding topology between IL-13 and IL-4 was observed. IL-13 shares a significant overlap in biological function with IL-4, a result of the common alpha chain component (IL-4Ralpha) in their respective receptors. Based on the available structural and mutational data, an IL-13/IL-4Ralpha model and a sequential mechanism for forming the signaling heterodimer is proposed for IL-13. PMID- 11419949 TI - Solution structure of interleukin-13 and insights into receptor engagement. AB - The complex and interrelated function of the interleukin cytokines relies on a range of pro-inflammatory and anti-inflammatory immune responses mediated by an array of receptors, and there is considerable cross-reactivity for related cytokines. Recent findings continue to elucidate the expression patterns of interleukin receptors associated with a range of diseases, including cancer. We report here the first experimentally determined high-resolution structure of human interleukin-13 (IL-13). The experimental structure is significantly different from an earlier homology model, which could have led to improper estimation of receptor interaction surfaces and design of mutational experiments. Similarities between the presented IL-13 structure and the homologous interleukin 4 (IL-4) are discussed. Additionally, mutation data for IL-4 and IL-13 are analyzed and combined with a detailed structural analysis of the IL-4/IL4Ralpha interface that leads us to postulate interactions at the IL-13/receptor interface. The structural comparison is used to interpret the different affinities for various receptors and establishes the basis for further mutational experiments and antagonist design. PMID- 11419950 TI - FUGUE: sequence-structure homology recognition using environment-specific substitution tables and structure-dependent gap penalties. AB - FUGUE, a program for recognizing distant homologues by sequence-structure comparison (http://www-cryst.bioc.cam.ac.uk/fugue/), has three key features. (1) Improved environment-specific substitution tables. Substitutions of an amino acid in a protein structure are constrained by its local structural environment, which can be defined in terms of secondary structure, solvent accessibility, and hydrogen bonding status. The environment-specific substitution tables have been derived from structural alignments in the HOMSTRAD database (http://www cryst.bioc. cam.ac.uk/homstrad/). (2) Automatic selection of alignment algorithm with detailed structure-dependent gap penalties. FUGUE uses the global-local algorithm to align a sequence-structure pair when they greatly differ in length and uses the global algorithm in other cases. The gap penalty at each position of the structure is determined according to its solvent accessibility, its position relative to the secondary structure elements (SSEs) and the conservation of the SSEs. (3) Combined information from both multiple sequences and multiple structures. FUGUE is designed to align multiple sequences against multiple structures to enrich the conservation/variation information. We demonstrate that the combination of these three key features implemented in FUGUE improves both homology recognition performance and alignment accuracy. PMID- 11419951 TI - The time scale of the catalytic loop motion in triosephosphate isomerase. AB - Loop 6 in the active site of Triosephosphate Isomerase (Saccharomyces cerevisiae) moves in order to reposition key residues for catalysis. The timescale of the opening and closing of this loop has been measured, at temperatures from -15 to +45 degrees C, using broadline solid state deuterium NMR of a single deuterated tryptophan located in the loop's N terminal hinge. The rate of the loop opening and closing was best detected using samples containing subsaturating amounts of a substrate analogue DL-glycerol 3-phosphate so that the populations of the open and closed forms were roughly equal, and using temperatures optimal for enzymatic function (30-45 degrees C). The T(2) values were much shorter than for unligated samples, consistent with full opening and closing of the loop at a rate of order 10(4) s(-1), and in good agreement with the rates estimated based on solution state 19F NMR. The loop motion appears to be partially rate limiting for chemistry in both directions. PMID- 11419952 TI - Solution-state NMR investigations of triosephosphate isomerase active site loop motion: ligand release in relation to active site loop dynamics. AB - Product release is partially rate determining in the isomerization reaction catalyzed by Triosephosphate Isomerase, the conversion of dihydroxyacetone phosphate to D-glyceraldehyde 3-phosphate, probably because an active-site loop movement is necessary to free the product from confinement in the active-site. The timescale of the catalytic loop motion and of ligand release were studied using 19F and 31P solution-state NMR. A 5'-fluorotryptophan was incorporated in the loop N-terminal hinge as a reporter of loop motion timescale. Crystallographic studies confirmed that the structure of the fluorinated enzyme is indistinguishable from the wild-type; the fluorine accepts a hydrogen bond from water and not from a protein residue, with minimal perturbation to the flexible loop stability. Two distinct loop conformations were observed by 19F NMR. Both for unligated (empty) and ligated enzyme samples a single species was detected, but the chemical shifts of these two distinct species differed by 1.2 ppm. For samples in the presence of subsaturating amounts of a substrate analogue, glycerol 3-phosphate, both NMR peaks were present, with broadened lineshapes at 0 degrees C. In contrast, a single NMR peak representing a rapid average of the two species was observed at 30 degrees C. We conclude that the rate of loop motion is less than 1400 s(-1) at 0 degrees C and more than 1400 s( 1) at 30 degrees C. Ligand release was studied under similar sample conditions, using 31P NMR of the phosphate group of the substrate analogue. The rate of ligand release is less than 1000 s(-1) at 0 degrees C and more than 1000 s(-1) at 30 degrees C. Therefore, loop motion and product release are probably concerted and likely to represent a rate limiting step for chemistry. PMID- 11419953 TI - Length analyses of mammalian G-protein-coupled receptors. AB - G-protein-coupled receptors (GPCRs) play a crucial role in mediating effects of extracellular messengers in a wide variety of biological systems, comprising the largest gene superfamily at least in mammals. Mammalian GPCRs are broadly classified into three families based on pharmacological properties and sequence similarities. These sequence similarities are largely confined to the seven transmembrane domains, and much less in the extracellular and intracellular loops and terminals (LTs). Together with the fact that the LTs vary considerably in length and sequence, the LT length of GPCRs has not been studied systematically. Here we have applied a statistical analysis to the length of the LTs of a wide variety of mammalian GPCRs in order to examine the existence of any trends in molecular architecture among a known mammalian GPCR population. Tree diagrams constructed by cluster analyses, using eight length factors in a given GPCR, revealed possible length relations among GPCRs and defined at least three groups. Most samples in Group J (joined) and Group M (minor) had an exceptionally long N terminal and I3 loop, respectively; and other samples were considered as Group O (other/original). This length-based classification largely coincided with the conventional sequence- and pharmacology-based classification, suggesting that the LT length contains some biological information when analysed at the population level. Principle component analyses suggested the existence of inherent length differences between loops and terminals as well as between extracellular and intracellular LTs. Wilcoxon rank transformation tests unveiled statistically significant differences between Group O and Group J, not only in the N-terminal and I3 loop, but also in the E3 loop. Correlation analyses identified an E1-I2 length-correlation in Group O and Group J and an N-E3 length-correlation in Group J. Taken together, these results suggest a possible functional importance of LT length in the GPCR superfamily. PMID- 11419954 TI - The antimicrobial properties of melanocytes, melanosomes and melanin and the evolution of black skin. AB - A biological issue that has not been satisfactorily resolved is the role of melanin in skin and other animal tissues. A hypothesis is outlined here to account for the evolution of black skin and the ubiquity of melanin in vertebrate tissues. Evidence is presented that melanization of skin and other tissues forms an important component of the innate immune defense system. A major function of melanocytes, melanosomes and melanin in skin is to inhibit the proliferation of bacterial, fungal and other parasitic infections of the dermis and epidermis. This function can potentially explain (a) the latitudinal gradient in melanization of human skin; (b) the fact that melanocyte and melanization patterns among different parts of the vertebrate body do not reflect exposure to radiation; (c) provide a theoretical framework for recent empirical findings concerning the antimicrobial activity of melanocytes and melanosomes and their regulation by known mediators of inflammatory responses. PMID- 11419955 TI - A logical analysis of the Drosophila gap-gene system. AB - This manuscript focuses on the formal analysis of the gap-gene network involved in Drosophila segmentation. The gap genes are expressed in defined domains along the anterior-posterior axis of the embryo, as a response to asymmetric maternal information in the oocyte. Though many of the individual interactions among maternal and gap genes are reasonably well understood, we still lack a thorough understanding of the dynamic behavior of the system as a whole. Based on a generalized logical formalization, the present analysis leads to the delineation of: (1) the minimal number of distinct, qualitative, functional levels associated with each of the key regulatory factors (the three maternal Bcd, Hb and Cad products, and the four gap Gt, Hb, Kr and Kni products); (2) the most crucial interactions and regulatory circuits of the earliest stages of the segmentation process; (3) the ordering of different regulatory interactions governed by each of these products according to corresponding concentration scales; and (4) the role of gap-gene cross-interactions in the transformation of graded maternal information into discrete gap-gene expression domains. The proposed model allows not only the qualitative reproduction of the patterns of gene expression characterized experimentally, but also the simulation and prediction of single and multiple mutant phenotypes. PMID- 11419956 TI - Structural changes in the neck linker of kinesin explain the load dependence of the motor's mechanical cycle. AB - The two-headed motor protein kinesin hydrolyzes ATP and moves on microtubule tracks towards the plus end. The motor develops speeds and forces of the order of hundreds of nanometers per second and piconewtons, respectively. Recently, the dependence of the velocity, the dissociation rate and the displacement variance on the load and the ATP concentration were measured in vitro for individual kinesin molecules (Coppin et al., 1997; Visscher et al., 1999) over a wide range of forces. The structural changes in the kinesin motor that drive motility were discovered by Rice et al. (1999). Here we present a phenomenological model for force generation in kinesin based on the bi-stable, nucleotide-dependent behavior of the neck linker. We demonstrate that the model explains the mechanical, kinetic and statistical (experimental) data of Coppin et al. (1997). We also discuss the relationship between the model results and experimental data of Visscher et al. (1999). PMID- 11419957 TI - Optimality under noise: higher memory strategies for the alternating prisoner's dilemma. AB - The Alternating Prisoner's Dilemma is a variant of the iterated Prisoner's Dilemma in which the players alternate in the roles of actor and recipient. We searched for strategies which are "optimal" in the Alternating Prisoner's Dilemma with noise (a non-zero probability that a player's decision will be transmitted incorrectly). In order to achieve success against a variety of other strategies, a strategy must be "self-cooperating" (able to achieve mutual cooperation with its clone), "C-exploiting" (able to exploit unconditional cooperators), and "D unexploitable" (able to resist exploitation by defectors). It must also have high evolutionary "dominance", a general measure of evolutionary performance which considers both resistance to invasion and the ability to invade other strategies. A strategy which meets these optimality criteria can evolve cooperation by invading a population of defectors and establishing a stable cooperative society. Most of the strategies commonly discussed in the Alternating Prisoner's Dilemma literature are low-memory strategies such as Tit For Tat, Pavlov, and Firm But Fair, but none of these strategies can simultaneously meet all of the optimality criteria. However, we discovered a class of higher memory "Firm Pavlov" strategies, which not only meet our stringent optimality criteria, but also achieve remarkable success in round-robin tournaments and evolutionary interactions. These higher memory strategies are friendly enough to cooperate with their clone, pragmatic enough to exploit unconditional cooperators, and wary enough to resist exploitation by defectors: they are truly "optimal under noise" in the Alternating Prisoner's Dilemma. PMID- 11419958 TI - Are sample sizes usually at least an order of magnitude too low for reliable estimates of leaf asymmetry? AB - Estimates of leaf size and asymmetry for individual trees are often obtained using sample sizes that are too small to take into account the possibility that size and asymmetry may be affected by the position of the leaf on the tree. This issue was addressed by exploring variation in leaf size and asymmetry within an individual of Alder (Alnus glutinosa). We found differences between branches for leaf size and for signed asymmetry but not for unsigned asymmetry. We also found that the size of a leaf was not correlated with its position on a branch and that the asymmetry of a leaf was not correlated with either its position on a branch or with the asymmetry of its neighbour. Repeated subsampling of a sample of 870 leaves showed that a subsample size approaching 500 leaves was required for consistently reliable estimates of the standard deviation of unsigned asymmetry. Smaller subsamples were required for consistently reliable estimates of mean unsigned asymmetry and of the mean and standard deviation of leaf size, but subsamples of less than 100 leaves provided consistently reliable estimates only of mean leaf size. For this species, reliable estimates of an individual's level of asymmetry are obtained only if several hundred leaves are sampled over several branches, but it is not necessary to sample the same sequence of leaves from each branch. PMID- 11419959 TI - Propionyl-L-carnitine as protector against adriamycin-induced cardiomyopathy. AB - Propionyl- l -carnitine (PLC) is a naturally occurring compound that has been considered for the treatment of many forms of cardiomyopathies. In this study, the possible mechanisms whereby PLC could protect against adriamycin (ADR) induced cardiomyopathy were carried out. Administration of ADR (3 mg kg(-1)i.p., every other day over a period of 2 weeks) resulted in a significant two-fold increase in serum levels of creatine phosphokinase, lactate dehydrogenase and glutamic oxaloacetic transaminase, whereas daily administration of PLC (250 mg kg(-1), i.p. for 2 weeks) induced non-significant change. Daily administration of PLC to ADR-treated rats resulted in complete reversal of ADR-induced increase in cardiac enzymes except lactate dehydrogenase which was only reversed by 66%. In cardiac tissue homogenate, ADR caused a significant 53% increase in malonedialdehyde (MDA) and a significant 50% decrease in reduced glutathione (GSH) levels, whereas PLC induced a significant 33% decrease in MDA and a significant 41% increase in GSH levels. Daily administration of PLC to ADR treated rats completely reversed the increase in MDA and the decrease in GSH induced by ADR to the normal levels. In rat heart mitochondria isolated 24 h after the last dose, ADR induced a significant 48% and 42% decrease in(14)CO(2)released from the oxidation of [1-(14)C]palmitoyl-CoA and [1 (14)C]palmitoylcarnitine, respectively, whereas PLC resulted in a significant 66% and 54% increase in the oxidation of both substrates, respectively. Interestingly, administration of PLC to ADR-treated rats resulted in complete recovery of the ADR-induced decrease in the oxidation of both substrates. In addition, in rat heart mitochondria, the oxidation of [1-(14)C]pyruvate, [1 (14)C]pyruvate and [1-(14)C]octanoate were not affected by ADR and/or PLC treatment. Moreover, ADR caused severe histopathological lesions manifested as toxic myocarditis which is protected by PLC. Worth mentioning is that PLC had no effect on the antitumour activity of ADR in solid Ehrlich carcinoma. Results from this study suggest that: (1) in the heart, PLC therapy completely protects against ADR-induced inhibition of mitochondrial beta -oxidation of long-chain fatty acids; (2) PLC has and/or induces a powerful antioxidant defense mechanism against ADR-induced lipid peroxidation of cardiac membranes; and finally (3) PLC has no effect on the antitumour activity of ADR. PMID- 11419960 TI - The influence of divalent cations on the analgesic effect of opioid and non opioid drugs. AB - It is generally accepted that divalent cations are involved in the nociceptive pathway. The effect of systemic co-administration of magnesium sulfate and calcium channel blockers (nifedipine, verapamil) on the analgesic effect of opioid (mixed mu/kappa: butorphanol) and non-opioid drugs (paracetamol) was investigated. Albino mice and rats were used as experimental animals. Magnesium sulfate and calcium channel blockers were given i.p., 30 min before the administration of butorphanol tartrate and paracetamol. Analgesia was measured using "hot-plate" ( 52.5( composite function)C), "tail-flick" (radiant heat source), "writhing" (acetic acid, 1%, i.p.) and "tail-clip" tests. The pain threshold was evaluated before and after the administration (i.p.) of the different agents. The effect of the combined administration of different agents on behavior, blood pressure and heart rate, was also determined. Nifedipine (5 mg kg(-1), i.p.) and verapamil (10 mg kg(-1), i.p.) potentiated the analgesic effect of butorphanol tartrate (0.25-2 mg kg(-1), i.p.) in all tests (synergism) and enhanced analgesic effect of paracetamol (50-125 mg kg(-1), i.p.), only in acetic acid writhing and tail-clip tests. Magensium sulfate (2.5 mg kg(-1), i.p.) potentiated the analgesic effect of butorphanol, but not that of paracetamol. Co administration of nifedipine and verapamil with either of butorphanol (0.25-2 mg kg(-1)) or paracetamol (50-125 mg kg(-1), i.p.) produced no significant effects on motor coordination, motor performance, locomotor activity, long-term memory or on the blood pressure and heart rate of experimental animals. Co-administration of magnesium sulfate, however, significantly induced sedation, inhibition of locomotor activity, motor performance and coordination, as well as impairing of long-term memory, as compared with butorphanol and paracetamol, administered alone. We conclude that the systemic co-administration of calcium channel blockers potentiated the analgesic effect of butorphanol against thermal, mechanical and chemical pain but enhanced that of paracetamol only against mechanical and chemical pain. Magensium sulfate enhanced the analgesic effect of butorphanol, but not that of paracetamol. These findings, merit further studies in animals and humans to evaluate the potential therapeutic benefits of such interactions. PMID- 11419961 TI - The effect of a single neonatal treatment (hormonal imprinting) with the antihormones, tamoxifen and mifepristone on the sexual behavior of adult rats. AB - Hormonal imprinting takes place perinatally during the first encounter between the hormone and its developing receptor. The presence of an excess of related molecules in that time provokes faulty (pathological) imprinting. In the present experiments single-neonatal treatment with 100 microg of tamoxifen completely abolished the adult male and female rats' sexual activity. Similar treatment with 100 microg of mifepristone (RU486) significantly enhanced the males sexual activity and non-significantly increased that of the females. The results demonstrate the importance of pathological imprinting during the perinatal development of sexual behavior. There are clear differences between the molecules having steroid (mifepristone) or non-steroid (tamoxifen) character, mediated through a ligand-receptor complex, and its effect in activating particular genes. PMID- 11419962 TI - Protective effect of L-arginine against stress-induced gastric mucosal lesions in rats and its relation to nitric oxide-mediated inhibition of neutrophil infiltration. AB - Pretreatment with l -arginine (150-600 mg kg(-1), i.p.), but not d -arginine (600 mg kg(-1), i.p.), protected against gastric mucosal lesions in rats with water immersion restraint stress over a 6-h period. This protective effect occurred in a dose-dependent manner. Increases in the activities of inducible nitric oxide synthase (iNOS) and myeloperoxidase (MPO), an index of tissue neutrophil infiltration, and the concentration of nitrite/nitrate, breakdown products of nitric oxide, and a decrease in the activity of constitutive nitric oxide synthase (cNOS) occurred in the gastric mucosal tissue with the development of gastric mucosal lesions. The l -arginine pretreatment attenuated the increases in iNOS and MPO activities and nitrite/nitrate concentration and the decrease in cNOS activity in the gastric mucosal tissue in a dose-dependent manner, while the d -arginine pretreatment did not. Both the protective effect of l -arginine (300 mg kg(-1)) against stress-induced gastric mucosal lesions and the attenuating effect of the amino acid on the increases in gastric mucosal iNOS and MPO activities and the decrease in gastric mucosal cNOS activity with the lesion development were counteracted by pretreatment with N(G)-monomethyl- l -arginine (100 mg kg(-1), s.c.), a nitric oxide synthase inhibitor, but not its d -isomer (100 mg kg(-1), s.c.). These results suggest that the protective effect of exogenously administered l -arginine against stress-induced gastric mucosal lesions in rats is, at least in part, due to nitric oxide-mediated inhibition of neutrophil infiltration into the gastric mucosal tissue. PMID- 11419963 TI - Bioequivalent or nonbioequivalent? AB - The current tests for bioequivalence are based on assumptions that are not valid in general; this paper shows why it is necessary to use a method that does not depend upon assumptions that cannot, and need not, be proved in general. PMID- 11419964 TI - Pharmacokinetics of two oral cyclosporin a formulations in clinically stable heart-transplant patients. AB - In order to evaluate the pharmacokinetics of cyclosporine A comparing the traditional (CsA-SCG) with the microemulsion formulation (CsA-ME), 20 clinically stable heart-transplant patients were enrolled in the study. All patients were on a thrice-daily dosage regimen (mean single dose 1.14 +/- 0.4 mgkg(-1)body weight) of CsA-SCG. The steady-state area under the concentration-time curve during a dosage interval was calculated during three sequential periods: (1) the first after the morning oral dose of CsA-SCG; (2) the second (8 days later) after 2 hours intravenous infusion of cyclosporine; (3) the third after the CsA-ME morning oral dose (30 days after the milligram-to-milligram dose conversion). After switching from standard formulation CsA-SCG to CsA-ME, significant changes were observed in C(max)(ss)( 732 +/- 178 vs 935 +/- 250 ng ml(-1), P< 0.001) and t(max)( 2.63 +/- 1.21 vs 1.36 +/- 0.49 h, P< 0.001). The CsA-ME mean bioavailability was higher than CsA-SCG ( 75 +/- 19 vs 66 +/- 16%;P< 0.001). The main CsA pharmacokinetic parameters of both formulations in clinically stable heart-transplant patients presented evident differences from data obtained in other transplant-patient populations. PMID- 11419965 TI - Improvement of RNA fingerprinting efficiency for the analysis of differential gene expression in human cardiac macro- and microvascular endothelial cells. AB - RNA fingerprinting by arbitrarily primed PCR (RAP-PCR) is a powerful tool to screen differential gene expression. However, PCR-based screening techniques show a high incidence of false positive results (40-90%). In order to increase the efficiency and feasibility of RAP-PCR, the original protocol was modified and applied to analyse differential gene expression in human coronary macro- (HCEC) and microvascular (HCMEC) endothelial cells. The major modifications introduced were: (i) the use of two primers for PCR amplification, instead of reverse transcription primer alone; (ii) the use of three cycles at low stringency followed by further amplification at high stringency; (iii) optimization of amplification cycle number, template amount, and concentration of primers, dNTP, Mg(2+); (iv) detection of fingerprints by silver staining; and (v) direct sequencing using RAP-PCR primers. Analysis of untreated and TNF alpha -stimulated (100 U ml(-1)for 1, 4, and 24 h) HCEC and HCMEC displayed 11 differentially expressed products by 18 primer combinations. Confirmation of results by RT-PCR showed that the rate of false positives attributable to our screening method was less than 20%. Among detected RAP-PCR products, the expression of Mn-superoxide dismutase, A20 zinc finger protein, and three novel genes (A/a, 4/d, 7/c) was more strongly modulated by TNF in HCEC than HCMEC. A further novel gene (B/e) was strongly expressed in HCMEC while only barely detectable in HCEC. In conclusion, modification of RAP-PCR strongly reduced the incidence of false positives, eliminated a radioactive requirement, and allowed sequencing without prior cloning, supplying an improved technology able to identify new differentially expressed genes between macro- and microvascular endothelial cells. PMID- 11419966 TI - Antioxidant action of bixin against cisplatin-induced chromosome aberrations and lipid peroxidation in rats. AB - Cisplatin is one of the most active cytotoxic agents in the treatment of cancer, but has serious side effects, inducing nephrotoxicity and chromosome aberrations. In this study we evaluated the role of the carotenoid bixin on cisplatin-induced oxidative stress in Wistar rats through three markers of oxidative damage: chromosome aberrations, glutathione depletion and lipid peroxidation. The animals were divided into six treatment groups with six rats in each (n= 6). The dose of cisplatin (5.0 mg kg(-1)body wt.) was injected i.p. and bixin (2.5 or 5.0 mg kg( 1)body wt.) was given by gavage at 48, 24 h and 10 min before the cisplatin injection. The treatment with the highest dose of bixin resulted in a statistically significant reduction, by about 33%, in cisplatin-induced abnormal metaphases (P< 0.05). A single dose of cisplatin enhanced the formation of lipid peroxides in 29% and resulted in a 29% depletion in renal glutathione 24 h after cisplatin administration (P< 0.05). The pretreatment with bixin reduced the total number of chromosome aberrations, inhibited the increase in lipid peroxidation, and inhibited renal glutathione depletion induced by cisplatin. Since the pretreatment with bixin alone was safe, under the present experimental conditions, the results suggest that bixin may have future clinical application after further studies. PMID- 11419967 TI - Infections in heterophile-negative patients. PMID- 11419969 TI - Physician satisfaction and emergency department laboratory test turnaround time. AB - OBJECTIVES: To determine the length of time for the components of the emergency department (ED) turnaround time (TAT) study in 1998 and to ascertain physician satisfaction concerning laboratory services to the ED. METHODS: Using forms supplied by the College of American Pathologists Q-Probes program, participants conducted a self-directed study of ED TAT over a 4-week period. Data requested included various times of day associated with the ordering, specimen collection, laboratory receipt, and result-reporting stages of stat ED TATs for potassium and hemoglobin. Additionally, practice-related questions associated with the laboratory were asked. Participating laboratories also provided a physician satisfaction survey for up to 4 physicians who were users of ED services. Results of both the TAT study and the physician satisfaction survey were returned by mail. Participants were drawn from the 952 hospital laboratories enrolled in the 1998 College of American Pathologists Q-Probes study on ED TAT. The main outcome measures included the components of the ED TAT process, factors associated with decreases in ED TAT, and the results of the physician satisfaction survey. RESULTS: Six hundred ninety hospital laboratories (72.4% response rate) returned data on up to 18 230 hemoglobin and 18 259 potassium specimens. Half of these laboratories responded that 90% of potassium tests were ordered and reported in 69 minutes or less, whereas the TAT for 90% of hemoglobin results was 55 minutes or less. Comparison of the components of TAT for both potassium and hemoglobin with similar studies done in 1990 and 1993 showed no change. Factors found to statistically contribute to faster TATs for both tests were laboratory control of specimen handling and rapid transport time. When whole blood specimens were used for potassium determination, TAT improved. Emergency department physicians chose the study-defined lower satisfaction categories of Often, Sometimes, Rarely, and Never for the questions concerning the laboratory being sensitive to stat testing needs (39.1%) and meeting physician needs (47.6%). Many of the physicians surveyed believed that laboratory TAT caused delayed ED treatment more than 50% of the time (42.9%) and increased ED length of stay more than 50% of the time (61.4%) when compared with other specialty users of the ED. CONCLUSIONS: Laboratory ED TATs have remained unchanged for almost a decade. Emergency department physicians are not satisfied with laboratory services. Although it appears that one issue may relate to the other, the interaction between the laboratory and the ED is quite complex and has been evolving for at least 30 years. Improvement in interoperability between the departments is essential for operational efficiency and patient care. Effective communication channels need to be established to achieve these goals. PMID- 11419970 TI - Computerized image analysis of p53 and proliferating cell nuclear antigen expression in benign, hyperplastic, and malignant endometrium. AB - CONTEXT: The endometrium is an intrinsically dynamic tissue with great capability for regeneration and proliferation; consequently, there is some overlap between features seen in benign, premalignant, and malignant lesions. This leads to marked intrabiopsy, interbiopsy, and interobserver variability. OBJECTIVE: We studied the specificity and sensitivity of computerized image analysis of molecular markers to evaluate its potential use as a diagnostic tool. DESIGN: Specimens from 100 patients were examined and the following histologic diagnoses were assigned: proliferative endometrium (n = 10), secretory endometrium (n = 10), endometrial hyperplasia (n = 40; 30 with no atypia, 10 with atypia), and carcinoma (n = 40; 20 endometrioid, 10 serous, and 10 clear cell). All cases were evaluated immunohistochemically for p53 and proliferating cell nuclear antigen (PCNA) expression. Computerized image analysis was performed with a CAS 200 digital analyzer. RESULTS: Expression of p53 was found only in carcinomas (65%) and endometrial hyperplasia with atypia (30%). Expression of p53 was higher in the poor prognostic categories (serous carcinoma and clear cell carcinoma) than in endometrioid carcinoma. In endometrioid carcinoma, p53 expression correlated with grade. Proliferating cell nuclear antigen showed a similar pattern of results to p53 in the various carcinoma subtypes and endometrioid carcinoma grades. Endometrial hyperplasia PCNA values were the lowest among all the groups. Both carcinomas and proliferative endometrium showed higher glandular and stromal PCNA values, significantly different from endometrial hyperplasia with atypia. In proliferative endometrium, stromal PCNA was the highest among all of the groups. The p53 and PCNA results correlated with each other for carcinoma. CONCLUSIONS: Computerized image analysis correlates well with the established morphologic groups of endometrial pathology and yields results consistent with previous studies. Owing to its higher degree of sensitivity, computerized image analysis is of potential use in cases of diagnostic dilemmas and can help objectively allocate the case in the correct category (e.g., proliferative endometrium vs. endometrial hyperplasia, endometrial hyperplasia with atypia vs. endometrioid carcinoma). It is particularly useful in the evaluation of stromal changes. PMID- 11419971 TI - Dural metastases. A retrospective surgical and autopsy series. AB - BACKGROUND: Metastases to dura constitute one of the least frequent and least studied patterns of neoplastic spread to the craniospinal axis. Older reports cited breast cancer as accounting for the largest number of dural metastases, often as a manifestation of end-stage disease. Since the early 1980s we have noted an increasing number and diversity of cancer types in patients with dural metastases in our surgical and autopsy neuropathology experience. Some surgical patients have experienced prolonged postoperative survival. METHODS: Review of autopsy records from 1982 to 1999 and surgical pathology records from 1990 to 2000 at a large university teaching hospital, including consultation files. RESULTS: Dural metastases removed at surgery (n = 33) were usually single, cranial, subdural lesions; coexistence with brain parenchymal or skull metastases varied according to tumor type. Unusual tumor types included carcinomas of colon, endometrium, cervix, and stomach. One well-documented patient developed postsurgical implantation metastases, and 2 patients had co-associated acute subdural hematomas. Surgical patients showed widely varying postoperative survival times, but in several of these patients survival times were in excess of 2 years. Dural metastases seen at autopsy (n = 27) could also be solitary and nodular, but more examples were extensive, diffuse, and both subdural and epidural in location. The expected cases of breast cancer (n = 5) occurred, but more examples of prostate cancer (n = 7) or unusual malignant neoplasms (cervical = 3, laryngeal = 1, gallbladder = 1, Ewing sarcoma = 1, intravascular lymphomatosis = 1, ocular melanoma = 1) were identified. CONCLUSIONS: This combined surgical and autopsy series shows a different distribution of tumor types causing dural metastases than older series. Postoperative outcome may be favorable in select, surgically treated patients. PMID- 11419972 TI - Evaluation of characteristics associated with acute splenitis (septic spleen) as markers of systemic infection. AB - CONTEXT: Acute splenitis, also known as acute splenic tumor or septic spleen, is loosely defined as a neutrophilic infiltrate and congestion within the red pulp accompanied by splenomegaly. Standard pathology texts state that the histologic finding of acute splenitis is reflective of septic states. However, this association has seldom been tested in the medical literature. OBJECTIVE: The purpose of this study was to determine if sepsis can be predicted by the presence of quantitative characteristics used to identify acute splenitis. METHODS: The postmortem splenic tissue of 20 clinically diagnosed cases of sepsis and 20 noninfectious control cases were retrospectively examined in a blinded fashion for amount of neutrophilic infiltration, weight (obtained from autopsy report), and presence of congestion. RESULTS: No significant correlation could be found between the parameters of neutrophilic infiltrate or splenic weight and clinically diagnosed septicemia. The presence of splenic congestion was unexpectedly found to be more likely with noninfectious causes of death. CONCLUSION: Acute splenitis is presently ill defined, and no available evidence convincingly shows that its presence suggests a septic state. PMID- 11419973 TI - The prognostic significance of DNA topoisomerase II-alpha (Ki-S1), p21/Cip-1, and p27/Kip-1 protein immunoexpression in oligodendrogliomas. AB - OBJECTIVE: To evaluate a possible association between clinical outcome of patients with oligodendroglioma and expression of 2 cyclin-dependent kinase inhibitors, p21/Cip-1 (p21) and p27/Kip-1 (p27), and of DNA topoisomerase II alpha (Ki-S1), which has been recently used as a marker of cellular proliferation. DESIGN: Ninety-one specially selected patients with cerebral oligodendrogliomas treated with surgery and radiotherapy were studied retrospectively. Tumor specimens were immunohistochemically examined with antibodies to p21, p27, and Ki-S1. A computerized color image analyzer was used to count immunostained nuclei. RESULTS: The mean Ki-S1 labeling index (LI) was found to be significantly prominent for World Health Organization (WHO) high grade tumors (9.5% vs. 3.2% for WHO low-grade tumors). In contrast, the mean p27 LI was significantly higher for low-grade tumors (43.3% vs 25.7% for high-grade tumors). The number of p21-positive cases and the mean p21 LI were found to be relatively equal for low- and high-grade tumors. For low-grade oligodendrogliomas, the progression-free and overall survival times were found to be significantly shorter for tumors with p27 LIs less than 20%. For high-grade oligodendrogliomas, survival times were significantly reduced for tumors with Ki S1 LIs greater than 10%. Regression-tree analysis identified 4 groups of oligodendrogliomas with distinctly different outcomes: (1) 32 patients with low grade tumors and p27 LIs greater than 20%; (2) 14 patients with low-grade tumors and p27 LIs less than 20%; (3) 25 patients with high-grade tumors and Ki-S1 LIs less than 10%; and (4) 20 patients with high-grade tumors and Ki-S1 LIs greater than 10%. CONCLUSIONS: Immunoreactivity for Ki-S1 and p27 was found to be useful for further subdividing oligodendroglioma prognoses among low-grade and high grade tumors. It seems unlikely that p21 immunohistochemistry will be of value for determining clinical outcomes for patients with oligodendrogliomas. PMID- 11419974 TI - Three-dimensional morphology of c-Kit-positive cellular network and nitrergic innervation in the human gut. AB - CONTEXT: -c-Kit-positive interstitial cells of Cajal (ICC) appear to play a key role in the normal motility function and development of intestine. Nitric oxide is considered to be the most important messenger of inhibitory nonadrenergic, noncholinergic nerves in the enteric nervous system. OBJECTIVES: The aims of this study were to examine the distribution of nitrergic innervation and ICCs in normal human bowel and to demonstrate interconnections between ICCs and nitrergic nerves and smooth muscle fibers using histochemical and immunohistochemical double-staining methods with a whole-mount preparation technique and confocal laser scanning microscopy. METHODS: Full-thickness small and large bowel specimens were obtained at autopsy from 18 children who died of nongastrointestinal diseases. A whole-mount preparation was performed for all specimens, and double staining was carried out with nicotinamide adenine dinucleotide phosphate (reduced form, NADPH)-diaphorase and c-Kit immunohistochemistry. Double immunofluorohistochemistry with neuronal nitric oxide synthase and c-Kit using confocal laser scanning microscopy was also performed in all specimens. RESULTS: The whole-mount preparation facilitated 3 dimensional visualization of the meshlike network of NADPH-diaphorase-positive nerve fibers in the myenteric plexus surrounded by a reticular network of c-Kit positive ICCs. The dense c-Kit-positive cellular network located between longitudinal and circular muscle layers and at the innermost part of circular muscle layer intermingled with the myenteric plexus. Short, fine processes of ICCs made connections with the muscle fibers and c-Kit-positive cells. CONCLUSIONS: The development of double-NADPH-diaphorase histochemistry and c-Kit immunohistochemistry staining technique in a whole-mount preparation provides an easy and useful method for investigating the association between c-Kit-positive cellular network and nitrergic neuronal network in the human bowel wall. The characteristic profiles of the c-Kit-positive cellular network and nitrergic neuronal network and their relationship with the smooth muscle fibers provide a morphologic basis for investigating intestinal motility disorders. PMID- 11419975 TI - National practice characteristics and utilization of pathologists' assistants. AB - OBJECTIVE: To obtain descriptive information regarding the practice characteristics and utilization of US pathologists' assistants. DESIGN: A self administered, mailed, voluntary, anonymous questionnaire was distributed to a cross-sectional sample of 515 US pathologists' assistants registered as members of the American Association of Pathologists' Assistants. The questionnaire contained items relating to subject demographics, practice characteristics, specific task performance, and amount of time spent per day on the performance of specific tasks. Descriptive statistics were used to describe the data in terms of measures of central tendency and dispersion. RESULTS: The response rate was 66.8%. The majority of questionnaires sent and received were from East Coast regions. Of all respondents, 46.6% were women, 57.6% were less than 40 years old, and 60.0% had been practicing 10 years or less. Over half (54.0%) had a master's degree. Almost the entire sample reported working 30 or more hours per week, with 43.4% reporting working more than 40 hours per week. The majority reported earning annual salaries between $56 000 and $75 000. Although task analysis of responses revealed a wide range of responses, the majority of the sample reported spending most of their daily time performing surgical specimen gross examinations (median 300 min/d). Approximately half of respondents also reported spending up to 90 minutes per day on nonspecific tasks such as logging specimens and answering the phone. Most respondents reported spending more daily time on such nonspecific tasks than on autopsy prosection or research. CONCLUSIONS: To our knowledge, this national survey provides the first description of pathologists' assistants across the United States. These data provide a useful tool for tracking changes in the profession. PMID- 11419976 TI - Characterization of microorganism identification in the United States in 1996. AB - CONTEXT: The National Inventory of Clinical Laboratory Testing Services (NICLTS) was designed to give an unbiased estimate of all patient testing performed by laboratories registered under the Clinical Laboratory Improvement Amendments in 1996. OBJECTIVE: Survey data were used to develop a profile of laboratory testing primarily intended to identify microorganisms or antibodies to these microorganisms. DESIGN: Estimates of the extent of microorganism identification were derived from the NICLTS database by identifying associated tests and methods. The volumes for tests performed at locations that primarily prepared blood components for distribution were excluded. Organisms of public health importance were identified from the National Notifiable Disease list maintained by the Centers for Disease Control and Prevention. PARTICIPANTS: Laboratories that were enrolled in the 1996 Online Certification Survey and Reporting System, maintained by the US Health Care Finance Administration, and that performed laboratory testing in 1996. OUTCOME MEASURE: Estimated volumes and associated confidence limits by test, method, specimen type, public health importance, and testing location. RESULTS: Excluding testing of the blood supply, 315 million tests (95% confidence limits, 280-354 million tests) were performed in the United States for microorganism identification. Those tests for which public health consensus requires national reporting represented 38% of this total. Although hospitals performed 46% of all microorganism identification, they only performed 33% of the testing for microorganisms of public health importance. Independent and specialty laboratories performed 38% of all testing but 65% of the testing for microorganisms of public health importance. Direct methods (methods not involving culture) were used in 77% of the tests for microorganisms of public health importance and in 42% of all identification tests. CONCLUSIONS: The distribution of microorganism identification testing found using NICLTS data is consistent with plans to modernize the public health surveillance system in the United States. PMID- 11419977 TI - Cytokeratin 7 and cytokeratin 20 in primary urinary bladder carcinoma and matched lymph node metastasis. AB - BACKGROUND: -Cytokeratin 7 (CK7) and cytokeratin 20 (CK20) are 2 types of intermediate filament protein. Expression of CK7 is seen in the majority of primary urinary bladder carcinomas. CK20 is restricted to superficial and occasional intermediate cells of the normal urothelium of the bladder. Aberrant CK20 expression has been documented in urothelial carcinoma and has proved useful as an ancillary diagnostic aid for urinary bladder tumor. Our hypothesis is that the pattern of CK7 and CK20 expression in metastatic urothelial carcinoma duplicates the expression of the same markers in the primary tumors. Therefore, immunohistochemical staining of metastatic tumors for these 2 markers may be helpful for differential diagnosis in ambiguous metastatic tumor deposits. OBJECTIVE: -To determine the concordance of CK7 and CK20 expression in primary bladder urothelial carcinoma and the matched lymph node metastasis. DESIGN: -We studied 26 patients with lymph node metastases who underwent radical cystectomy and bilateral lymphadenectomy for bladder carcinoma. Immunohistochemical staining for CK7 and CK20 was performed on formalin-fixed paraffin-embedded tissues containing primary cancers and lymph node metastases. RESULTS: -In all cases, there was a concordant expression of CK20 in the primary cancer and its matched lymph node metastasis. Twelve cases (46%) showed positive CK20 immunoreactivity in the primary tumor and its matched lymph node metastases, whereas 14 cases (54%) were negative for CK20 in both the primary tumor and lymph node metastasis. All cases showed positive CK7 immunoreactivity in the primary cancers and matched lymph node metastases. CONCLUSIONS: -CK20 immunoreactivity is reliably observed in metastases from bladder cancer when the primary tumor expresses CK20. PMID- 11419978 TI - Declining clinical autopsy rates versus increasing medicolegal autopsy rates in Halifax, Nova Scotia. AB - The downward trend in the rate of clinical autopsies has been extensively documented in the literature. This decline is of concern when the benefits of the clinical autopsy are considered. In contrast, the rate of medicolegal autopsies has not been studied in such detail. What little reference there is to medicolegal autopsy rates suggests an absence of the same downward trend. A retrospective review of autopsy data over a 13-year period from the Queen Elizabeth II Health Sciences Centre in Halifax, Nova Scotia, and from the Office of the Chief Medical Examiner of Nova Scotia was conducted. This review showed a difference between the rates of clinical and medicolegal autopsies for the metro Halifax area. The clinical autopsy rate was consistently less than 30% and declined to 15% in 1999, while the medicolegal autopsy rate was consistently greater than 40% and rose to 62% in 1999. The literature proposes many reasons for the decline in the clinical autopsy rate, but none for this difference between rates. The explanation proposed here is the changing and currently uncertain purpose of the clinical autopsy versus the clear, and consistent over time, purpose of the medicolegal autopsy. PMID- 11419979 TI - Infantile hemangioendothelioma of the liver in an adult. AB - Hepatic infantile hemangioendothelioma (IHE) is a rare benign vascular tumor of the liver found mostly in infancy and early childhood. We describe herein a tumor of the liver, morphologically indistinguishable from IHE, that presented in an adult. The patient is a 56-year-old woman with a history of ductal carcinoma in situ of the left breast and a mass in the liver. The patient underwent resection of the tumor, and after 23 months of follow-up she is alive with no evidence of expansion of the liver tumor. PMID- 11419980 TI - Pulmonary valve papillary fibroelastoma. A case report and review of the literature. AB - Papillary fibroelastomas are rare and benign cardiac tumors that typically affect the cardiac valves. To the best of our knowledge, the English literature contains only 1 case report of pulmonary valve fibroelastoma diagnosed by echocardiogram and confirmed by surgical resection. There is a paucity of pathology literature on this subject. We describe an additional case of pulmonary valve fibroelastoma diagnosed by transesophageal echocardiography and magnetic resonance imaging confirmed by pathologic examination in a patient who also had a thymoma. PMID- 11419981 TI - Papillary carcinoma of thyroid metastatic to the pituitary gland. AB - We describe a 35-year-old woman who presented with diabetes insipidus caused by metastatic papillary carcinoma of the thyroid involving the pituitary gland, 25 years after treatment for a papillary carcinoma of thyroid and 17 years after treatment for multiple pulmonary metastases. The literature contains 10 previously described cases of metastatic thyroid carcinoma involving the sella, but only 2 of these cases had unequivocal metastases to the pituitary gland, making the present case, to our knowledge, the third reported case of unequivocal hematogenous metastasis of thyroid carcinoma to the pituitary gland. The pituitary tumor was removed by transsphenoidal surgery, and the tissue was examined by conventional histology, extensive immunohistochemistry, and electron microscopy. The findings confirmed the tumor to be papillary thyroid carcinoma. To our knowledge, this is the first report citing use of thyroid transcription factor-1 to establish a thyroid source of a pituitary metastasis. PMID- 11419982 TI - Vertebral metastases from pineoblastoma. AB - Two male patients aged 26 and 18 years presented with vertebral metastases originating from pineoblastomas on which surgery had been performed 8 years and 5 months earlier, respectively. In the first case in which the metastasis developed in the T8 corpus, the disease is presently under control after high-dose chemotherapy and autologous blood stem cell transplantation. The second patient (sacral metastasis), despite aggressive adjuvant therapy, died 2 years after the last operation because of spinal seeding. These uncommon cases are discussed with reference to the literature on extraneural metastases that originate from neuroepithelial tumors of the central nervous system. PMID- 11419983 TI - Carbohydrate antigen 19-9 and carcinoembryonic antigen immunostaining in benign multicystic mesothelioma of the peritoneum. AB - A 58-year-old Italian man was incidentally discovered to have an elevated carbohydrate antigen 19-9 (CA-19-9) level of 132 U/mL on routine blood testing. Multisystem imaging studies revealed multiple benign-appearing cysts of the liver and single cysts in the pancreas and kidney parenchyma. Throughout 14 months, fluctuations were observed in the elevated serum CA-19-9 levels from 99 to 450 U/mL. Serum carcinoembryonic antigen (CEA) and other tumor markers were normal. Laparoscopy disclosed multiple cystic lesions on the surface of the liver, on the serosal surface of the ileum, and in the mesentery. Electron microscopy characterized the cells as mesothelial. The pathologic diagnosis was benign multicystic mesothelioma of the peritoneum (BMMP). Aspirated fluid from the liver cyst revealed CA-19-9 levels at 28 500 U/mL, strongly linking the elevated serum CA-19-9 levels with mesothelial cyst secretion. Immunostaining was positive for CA-19-9, CEA, and cancer antigen 125 (CA-125). We believe this is the first documented instance of CA-19-9 and CEA secretion in BMMP. PMID- 11419984 TI - Myeloperoxidase-positive intravascular large B-cell lymphoma. AB - Intravascular large B-cell lymphoma (IVLBL) is an uncommon form of non-Hodgkin lymphoma that is also known as malignant angioendotheliosis, intravascular lymphomatosis, and angiotropic large-cell lymphoma. The disease is characterized by a bizarre population of neoplastic cells, which are found systemically within vascular lumina. Although originally thought to be a neoplastic process of the endothelial cells, it has since been demonstrated, by molecular techniques and immunohistochemistry, that the neoplastic cells are of lymphoid origin. The differential diagnosis of these lesions includes granulocytic sarcomas that can be distinguished from IVLBL or other lymphomas by the presence of immunohistochemical positivity for myeloperoxidase. We describe a patient with a history of a myelodysplastic syndrome who subsequently developed IVLBL, which demonstrated immunohistochemical positivity for myeloperoxidase. To our knowledge, this represents the first case of a malignant lymphoma to demonstrate such findings. PMID- 11419985 TI - Low-grade B-cell lymphoma with coexpression of both CD5 and CD10. A report of 3 cases. AB - The coexpression of CD5 and CD10 has previously been reported in cases of intermediate- and high-grade lymphomas and in precursor B cells in normal or regenerating bone marrow. We report 3 cases of low-grade B-cell lymphoma that were found to coexpress CD5 and CD10 at the time of initial diagnosis. The first case was classified as small lymphocytic lymphoma; the second as follicle center lymphoma, follicular grade 1; and the third as small B-cell lymphoma otherwise not specified. Currently, the clinical implication of the coexpression of CD5 and CD10 is not known. We describe this finding to highlight the difficulty that may be encountered in classifying lymphomas in cases where this coexpression is present. PMID- 11419986 TI - Allergic granulomatosis secondary to a limited form of Churg-Strauss syndrome. AB - Allergic granulomatosis is a disorder of obscure etiology characterized by infiltration of lymph nodes with histiocytic granulomas and eosinophils. In this report, we describe a case of allergic granulomatosis that is consistent with Churg-Strauss disease limited to lymph nodes. The histologic findings of this patient's lymph nodes demonstrated eosinophilic abscesses and ring-shaped necrotizing and nonnecrotizing granulomas surrounding hyperplastic germinal centers. We report herein a rare type of reactive lymphadenopathy and present its histologic, immunohistochemical, and flow cytometric findings, which may allow its distinction from other lymphadenopathies. PMID- 11419987 TI - CD79a(+) T-cell lymphoblastic lymphoma with coexisting Langerhans cell histiocytosis. AB - Although there is a close association between Langerhans cell histiocytosis and malignant neoplasms, simultaneous occurrence of lymphoblastic lymphoma and Langerhans cell histiocytosis in the same lymph node is an extremely rare finding. Herein, we describe such a case in a 26-year-old woman who presented with progressive cervical lymphadenopathy. The lymphoma cells have an immature T cell phenotype (terminal deoxynucleotidyl transferase(+), HLA-DR(+), CD34(+), CD38(+), and CD7(+)) with expression of both CD3 and CD79a on immunohistochemical stain. The Langerhans cells are present focally with the characteristic morphologic features and immunophenotype (CD1a(+) and S100(+)). The significance of CD79a coexpression in T-cell lymphoblastic lymphoma and the association between lymphoblastic lymphoma and Langerhans cell histiocytosis are discussed. PMID- 11419988 TI - Dura mater involvement in ochronosis (alkaptonuria). AB - Clinical manifestations of alkaptonuria have been well described and are most commonly characterized by ochronosis or pigmentation of connective tissue. Sites most commonly involved in ochronosis include joints, heart, skin, and kidney. We describe a 66-year-old woman with a history of alkaptonuria who had widespread ochronosis. The dura mater showed extensive pigment deposition, which was evident both grossly and microscopically at autopsy. To our knowledge, description of dura mater involvement by ochronosis has not been previously reported in the literature. PMID- 11419989 TI - Pathologic quiz case. Large posterior mediastinal mass in a young woman. Pathologic diagnosis: localized hyaline-vascular-type Castleman disease (angiofollicular lymphoid hyperplasia). PMID- 11419990 TI - Pathologic quiz case. Elderly man with bright red blood per rectum. Pathologic diagnosis: metastatic hepatic epithelioid angiosarcoma. PMID- 11419991 TI - Pathologic quiz case. Large retroperitoneal mass in an asymptomatic 43-year-old woman. Pathologic diagnosis: solid and papillary epithelial neoplasm of pancreas. PMID- 11419992 TI - Pathologic quiz case. Intranuclear inclusions in allograft kidney. Pathologic diagnosis: human polymavirus-associated interstitial nephritis in the allograft kidney. PMID- 11419993 TI - Pathologic quiz case. Chronic pneumonia after gastrectomy. Pathologic diagnosis: chronic aspiration lipoid pneumonia with Mycobacterium abscessus. PMID- 11419994 TI - Pulmonary nocardiosis in a patient with human immunodeficiency virus. PMID- 11419995 TI - Chlamydia trachomatis in a thin-prep Papanicolaou Test. PMID- 11419996 TI - Building social relationships through valued roles: three older adults and the community membership project. AB - The experiences of three older adults with developmental disabilities highlight ways in which social relationships can be nourished through the establishment of community roles. Members of the Community Membership Project of the Center on Aging and Community used paid community builders working individually with older persons who had developmental disabilities in order to help them obtain positive, valued social roles and relationships with nondisabled community members. As a means to this end, participants were supported to engage in community activities corresponding to the interests and talents discovered in an initial period of exploration. Stories of these community builders' work illustrate the intentional strategies and concerted effort necessary to create community connections and meaningful relationships. PMID- 11419997 TI - Continuity or discontinuity of family involvement following residential transitions of adults who have mental retardation. AB - Family involvement with adults who have mental retardation following a residential transition to a nonparental living situation was examined. We found that aging mothers were highly involved in the relocation process and had frequent contact and continued emotional involvement with their adult child. Mothers became increasingly satisfied with their level of contact with their child over time, less worried about the future, and had decreasing levels of direct caregiving and contact with residential staff. Adult siblings reported improved sibling relationships over time. Siblings whose brother or sister moved out of the parental home increased their shared activities and felt less pessimistic about the future. Findings address a critical gap in knowledge about the life course roles of families of persons with mental retardation. PMID- 11419998 TI - Finding a Voice: Poetry and People With Developmental Disabilities. AB - Poetry may be used as a vehicle for self-expression with individuals who have developmental disabilities, even with those who may be only minimally verbal. Poetry by well-known poets may be used to illustrate concepts and to enhance the creative experience. Then, through strategic questioning, participants' ideas are recorded in poetic form. The results may be therapeutic in many ways as well as powerful expressions of human experience. PMID- 11419999 TI - Social networks of people with mental retardation in residential settings. AB - Information was collected on the social networks of 500 adults with mental retardation receiving different types of residential supports. Results indicated that (a) the reported median size of participants' social networks (excluding staff) was 2 people; (b) 83% of participants were reported to have a staff member; 72%, a member of their family; 54%, another person with mental retardation; and 30%, a person who did not fit into any of these categories in their social network; (c) variation in the size and composition of participants' social networks was associated with a range of variables, including the personal characteristics of residents (age, autism, ability, and challenging behavior), the type of previous and current accommodation, staffing ratios, institutional climate, and the implementation of "active support." PMID- 11420000 TI - Should adults with phenylketonuria have diet treatment? PMID- 11420001 TI - University affiliated programs as specialty medical providers and the emerging managed-care system. PMID- 11420002 TI - Disney's Tarzan, Edgar Rice Burroughs' Eugenics, and Visions of Utopian Perfection. PMID- 11420003 TI - Editor's Response. PMID- 11420004 TI - How do you understand "learning difficulties"? Towards a social theory of impairment. PMID- 11420005 TI - The Troubling Interrogation of Travis Lee Wright. PMID- 11420006 TI - "Free Appropriate Public Education: The Law and Children with Disabilities." PMID- 11420007 TI - Leveraging Federal Funding in the States to Address Olmstead and Growing Waiting Lists. PMID- 11420008 TI - Imaging of cartilage and mimic muscles with MRI: anatomic study in healthy volunteers and patients with unilateral cleft lip and palate. AB - OBJECTIVE: The aim of the study was to visualize different soft tissues in the perioral, nasal, and paranasal region by means of magnetic resonance imaging (MRI) in patients with unilateral cleft lip and palate (UCLP). DESIGN: In this descriptive study, images of different MRI systems were assessed and compared. METHOD: MRI was applied in five consecutive patients operated on for UCLP who underwent secondary lip and nasal correction, two patients who had not had UCLP operations, and five healthy volunteers as controls. The mimic muscles, vessels, and nasal cartilages were evaluated. RESULTS: It was possible to visualize different parts of the paranasal and perioral mimic muscles and their interlacement in the upper lip. The nasal cartilages were also visible, and the changes after operation were demonstrated. CONCLUSIONS: MRI shows differentiated visualization of soft tissues in the cleft region and their changes after surgery. It is a valuable tool in the preoperative planning and postoperative follow-up in patients with UCLP. PMID- 11420009 TI - A method for three-projection infant cephalometry. AB - OBJECTIVE: To assess morphology and growth in infants and children with craniofacial anomalies based on comprehensive digitization of radiographic films in three, mutually orthogonal projections. METHOD: The method consists of (1) acquisition of radiographic films in a highly standardized three-projection (lateral, frontal, and axial) cephalometer, (2) marking and digitization of a total of 279 anatomical landmarks in the three projections, and (3) computation and presentation (tabular and graphical) of 356 linear and angular variables describing the craniofacial morphology, including soft tissue. Computation of statistical entities describing a patient, a group of patients, the differences between patients or groups of patients was carried out. Error assessment of the method involved investigation of error distribution among a number of error sources. Duplicate digitization of radiographic films from 30 randomly selected patients, and from 10 dry skulls, was carried out to determine the errors contributed by the procedure of landmark digitization and the distribution of error among landmarks and variables, as well as between projections. RESULTS: The average error due to landmark digitization, s(i), determined by duplicate digitization and calculated by use of Dahlberg's formula was 0.8 mm for linear variables and 1.6 degrees for angular variables. CONCLUSION: This method of infant cephalometry has been shown to be highly accurate and reproducible, and it adds significant new potential for, e.g., asymmetry detection, population comparison, and growth measurements compared to other cephalometric techniques due to its standardized acquisition and digitization protocol, inclusion of an axial projection, and the large number of well-defined landmarks and variables involved. PMID- 11420010 TI - Facial reconstruction on the abnormal skull model of a living patient. AB - OBJECTIVE: The concept of using a skull as the armature upon which to reconstruct a face has been understood for many years. The objective of this study was to blindly test this technique by using a skull that was reconstructed from the computed tomography (CT) scans of a living 48-year-old man. DESIGN: The patient had a bilateral cleft lip, alveolus, and palate. A model of the complete skull produced by a computer-controlled milling machine that used data from the CT scans was used for the reconstruction. Surgery was then performed, and new dental prostheses were also constructed. After surgery, a second model of the relevant part of the skull was made and the reconstruction modified accordingly. CONCLUSION: Despite having no information about the patient apart from that which could be gleaned from the model skull, the reconstructions demonstrate the approximate appearance of the face before and after surgery. Even without the extra information routinely available in forensic cases, a face broadly similar to that of the living patient could be achieved. PMID- 11420011 TI - Long-term evidence for favorable midfacial growth after delayed hard palate repair in UCLP patients. AB - OBJECTIVE: To investigate whether delayed hard palate repair resulted in better midfacial growth in the long term than previously achieved with "conventional" surgical methods of palatal closure. DESIGN AND SETTING: Long-term cephalometric data from patients with unilateral cleft lip and palate were available from two Scandinavian cleft centers. The patients had been treated by different regimens, particularly regarding the method and timing of palatal surgery. Patients were analyzed retrospectively, and one investigator digitized all radiographs. PATIENTS: Thirty consecutively treated subjects from each center, with cephalograms taken at three comparable stages between 10 and 16 years of age. RESULTS AND CONCLUSIONS: Patients whose hard palates were repaired late (early soft palate closure followed by delayed hard palate repair at the stage of mixed dentition) had significantly better midfacial development than patients in whom the hard palate was operated on early with a vomer flap, and then during the second year of life, the soft palate was repaired with a push-back procedure. As the growth advantage in the delayed hard palate repair group was accomplished without impeding long-term speech development, the delayed repair regimen proved to be a good alternative in surgical treatment of patients with unilateral cleft lip and palate. PMID- 11420012 TI - Incisive suture (fissure) in the human fetus: radiographic and histologic study. AB - OBJECTIVE: An anatomic study, in man, of the structure commonly known as the incisive suture (sutura incisiva) or incisive fissure has been performed to determine whether this structure belongs to the morphofunctional concept of a facial suture. MATERIAL: Eighteen palates of human fetuses of 9 weeks to 29 weeks after conception. METHOD: Anatomic, radiographic examination of the palate. Histologic study of sagittal and parasagittal sections of the palates (3 mm each). RESULTS: The histologic aspect is that of a facial suture, with very poor vascularization. This suture is partial, limited laterally by osseous trabeculae. CONCLUSIONS: This particular suture, whose function in growth of the palatal process of the maxilla is discussed, could represent the phylogenetic vestige of the incisive-maxillary suture present in all nonhuman mammals. PMID- 11420013 TI - Food and Drug Administration regulation of orthotic cranioplasty. AB - OBJECTIVE: To present information regarding the current federal regulation of cranial orthotics used for the treatment of deformational plagiocephaly as well as to discuss concerns raised by the Food and Drug Administration regarding the safety and effectiveness of these devices. BACKGROUND: Although first introduced in 1979, the use of orthotic helmets for the treatment of deformational plagiocephaly was slow to gain acceptance. However, with the recent increase in infants presenting with this condition, numerous orthotic treatment programs have been established throughout the country. Until recently, federal regulation of this "industry" was largely ignored. REGULATION: In 1995 our office was served notice that our orthosis would require clearance from the FDA. Since the FDA had never approved a medical device of this kind, clearance presented a significant challenge. However, after 3 years of providing clinical data, clearance was finally granted, and a new device category known generically as "cranial orthosis" was created. A cranial orthosis is considered to be a Class II neurology device and requires both general and special controls in order to ensure its safety and effectiveness. SUMMARY: Orthotics used for the treatment of deformational plagiocephaly are regulated by the FDA and are considered Class II neurology devices. Submission of a premarket notification (510[k]) is required prior to placing these devices on the market. PMID- 11420014 TI - Patterns of body and visceral growth in human prenates with clefts of the lip and palate. AB - OBJECTIVE: To address the hypothesis that human prenates with birth defects such as clefts of the lip and palate show growth patterns in which body size and visceral growth are aberrant for prenatal age. METHODS: Body size and organ weight measurements were derived from a cross-sectional population sample of 167 legally donated human embryos that were representative of key periods in visceral organogenesis and growth. Measurements included those of overall body size (i.e., length and weight) and weights of major organs. Data were analyzed for two comparative (age-matched) groups consisting of 120 typical-for-age "control" and 47 specimens showing the three typical cleft types. Organ weight data for the entire cleft group and for each of the three cleft subgroups were compared with those of the control group. RESULTS: Cleft specimens showed crown-rump body sizes that were within one standard deviation of the control group. Among the entire cleft sample, each of the organ weight values was generally consistent with the organ weights observed for the age-matched control group. When organ weights for the cleft group differed from the controls, with one exception, some organs showed either increased or decreased organ weights. One remarkable exception was with lung weights, which were reduced for each of the prenates across the three cleft types. Among the three cleft groups, fetuses with cleft lip and palate showed the highest percentage of organ weight discrepancies (i.e., smaller than expected) as compared with the group with cleft palate, which showed the lowest. CONCLUSIONS; Organ weight aberrancies were observable (whether increased or reduced weights), and such aberrancies were not apparent until approximately 24 weeks (fertilization age). PMID- 11420015 TI - Investigations of a pressure-sensitive theory of marginal velopharyngeal inadequacy. AB - PURPOSE: This two-part project was designed to test a pressure-sensitive theory of marginal velopharyngeal inadequacy (MPVI). Specifically, are select subgroups of children with MPVI perceived as hypernasal because they fail to achieve consistent closure during vowels and semivowels while demonstrating adequate closure during pressure consonants? METHODS: In part one, 36 children with cleft palate and other craniofacial anomalies were examined using a clinical assessment protocol that included nasometry and perceived ratings of hypernasal resonance. Children with nasalance percentages above threshold during low-pressure (LP) productions and below threshold for high-pressure (HP) productions were placed in one group (group 1), while children with nasalance percentages below threshold for both LP and HP sentences were placed in another (group 2). Children in the two groups were age- and sex-matched. In part two, endoscopic data were examined for 10 additional children who received nasometry, perceived hypernasal resonance scores, and videoendoscopy on the same day and who received higher mean nasalance measures during production of LP sentences than during production of HP sentences. RESULTS: The results of part one confirmed that children in group 1 were perceived as being significantly more hypernasal than children in group 2 (mean(group 1) = 2.17, mean(group 2) = 1.50; t = 2.75, p =.01). However, results of endoscopic testing failed to demonstrate a consistent observable physiologic pattern of velopharyngeal inadequacy that would confirm the theory that some patients with MVPI are perceived as being hypernasal because of difficulty achieving velopharyngeal closure during vowels and semivowels. CONCLUSIONS; The findings provide partial support for a pressure-sensitive theory of MVPI and demonstrate the value of using both HP and LP sentences to evaluate patients with MVPI. PMID- 11420016 TI - Nasal airflow patterns during the velopharyngeal closing phase in speech in children with and without cleft palate. AB - OBJECTIVES: (1) To study the nasal airflow patterns during the velopharyngeal closing phase in speech produced by children with and without cleft palate. (2) To compare the nasal airflow patterns in bilabial, dental, and velar articulation in these children. DESIGN: Prospective, cross-sectional study of a consecutive series of children with cleft palate referred for routine speech evaluation and controls. SETTING: Sahlgrenska University Hospital, Goteborg, Sweden. PARTICIPANTS: Seventeen children with cleft lip and palate or cleft palate only and 22 controls aged 7 and 10 years. METHOD: Nasal airflow was transduced with a pneumotachograph attached to a nose mask and registered together with the acoustic speech signal. Sentences containing nasal-to-stop combinations in bilabial, dental, and velar articulatory positions were used. MAIN OUTCOMES MEASURES: The duration from peak to 5% nasal airflow, the maximum flow declination rate, and the nasal airflow at selected points in time during the transition from nasal-to-stop consonants. RESULTS: In the cleft palate group, duration from peak to 5% nasal airflow was clearly longer than among the controls (p <.0001). The declination of airflow was slower (p <.006) and the rate of nasal airflow at the release of the stop consonant was higher (p <.004) in the cleft palate group. Differences between bilabial versus dental and velar articulation were found in the control group. CONCLUSION: Studies of the temporal and dynamic characteristics of the nasal airflow variations during speech appear potentially useful for the assessment of velopharyngeal function. PMID- 11420017 TI - Hypodontia and tooth formation in groups of children with cleft, siblings without cleft, and nonrelated controls. AB - OBJECTIVE: To compare the occurrence of hypodontia, dental age, and asymmetric dental development in children with cleft with their siblings and a nonsibling control group. SUBJECTS: The sample consisted of 54 children with cleft (aged 4.2 to 13.1 years), who had at least one sibling available for an orthopantomogram, 63 siblings without cleft (aged 4 to 14.9 years) and 250 controls without cleft (aged between 4 and 14.9 years). METHOD: Hypodontia, dental age, and asymmetric dental development were assessed on panoramic radiographs of the children with cleft, the siblings without cleft, and the control children without cleft. RESULTS: Both the cleft (p <.001) and the sibling group (p <.05) showed a significantly higher frequency of hypodontia and a significantly higher occurrence (cleft p <.01 and sibling group p <.001) of asymmetric dental development, compared with the control group. Only a small, but insignificant, delay in dental development could be found in the cleft and the sibling group. CONCLUSIONS; The cleft subjects with siblings showed a significantly higher occurrence of hypodontia and asymmetric dental development than the noncleft control group. This may suggest a genetic component for the occurrence of hypodontia and asymmetric dental development. PMID- 11420018 TI - Quality of life in adults with repaired complete cleft lip and palate. AB - OBJECTIVE: This study evaluated the quality of life in adult Swedish subjects with repaired complete cleft lip and palate (CLP). DESIGN/PATIENTS: Sixty-eight adults with repaired CLP were compared with a group of 66 adults without cleft matched by gender and age. OUTCOME MEASURES: The outcome measures included a self report questionnaire concerning quality of life in general, well-being, and health-related quality of life. RESULTS: The CLP group rated their quality of life significantly lower than did the control group in the areas of life meaning (p <.05), family life (p <.0001), and private economy (p <.01). There were no significant differences between the groups concerning well-being. In the CLP group, the health-related quality of life was significantly lower concerning global life (p <.0001), disturbance to life (p <.01), well-being (p <.0001), social contacts (p <.0001), and family life (p <.05) but significantly higher concerning ability to make the most of leisure time (p <.001) and to be active (p <.001). CONCLUSIONS: The CLP group perceived a marked impact of the handicap on their lives concerning global aspects, well-being, and social life. More practical and tangible aspects of their daily living, however, were not affected, and only some minor aspects of their quality of life in general were poor in comparison with the control group, which indicates a fairly good life adjustment in spite of the handicap. PMID- 11420019 TI - Facial attractiveness and facial impairment ratings in children with craniofacial malformations. AB - OBJECTIVE: This study examined (1) the effects of type of malformation, sex of ratee, and sex of rater on facial attractiveness and facial impairment ratings, and (2) the reliability of judgments on facial attractiveness and facial impairment and the association between these two constructs. SETTING: A university hospital for children. PARTICIPANTS: Raters were eight volunteers from the student population in a university, four men and four women. MAIN OUTCOME MEASURES: Raters judged frontal and lateral view slides of children with various types of craniofacial malformations both on a 5-point facial attractiveness scale and on a 5-point facial impairment scale. RESULTS: Main effects were found for type of malformation, sex of ratee, and sex of rater. No interaction effects were found among type of malformation, sex of ratee, and sex of rater. Interrater reliability was moderate to high, both for attractiveness ratings and for impairment ratings. The correlation between facial attractiveness and facial impairment was also moderate to high. CONCLUSIONS; Both condition parameters (type of malformation) as well as social parameters (sex of rater and sex of ratee) seem to influence judgments on attractiveness and impairment. Facial attractiveness and facial impairment can be rated reliably in children with (cerebro)craniofacial dysplasias. Raters consider these concepts to be very similar but not identical. PMID- 11420020 TI - Is it possible to prevent cleft palate by prenatal administration of folic acid? An experimental study. AB - OBJECTIVE: In this study, folic acid was tested for its antiteratogenic effects on experimentally induced cleft palate in animals. DESIGN: Eleven pregnant Lew 1 A dams (75 fetuses) received 200 mg/kg procarbazine via gastric tubing on postconception (p.c.) day 14 to induce a cleft palate (CP); seven of the pregnant dams (45 fetuses) were additionally given 4 mg/kg folic acid subcutaneously from the 14th to the 17th day p.c. As a control group, three more pregnant dams (24 fetuses) were not treated with the drugs mentioned above. All fetuses were delivered by Caesarian section on day 20 p.c. OUTCOMES MEASURED: All fetuses were weighed and examined macroscopically with a stereomicroscope. Each fetal head was cut into 35 frontal sections and scrutinized histologically. RESULTS: None of the control fetuses (n = 24) exhibited a cleft. Without folate administration, 90% of the fetuses (27 of 30) that received procarbazine exhibited a CP. After additional prenatal folate administration, this rate remained virtually unchanged (91%; 41 of 45). However, the proportion of complete (total) CP (4%) was significantly (p <.0001) lower than in the group without folate (53%). Cleft associated microgenia and microglossia were also significantly less frequent when folate was administered prenatally: microgenia was reduced by 22% (p =.029) and microglossia by 24% (p =.032). CONCLUSIONS: On the basis of these results, folate has a partial ameliorating effect on the teratogenicity of procarbazine given to pregnant rats. Additional studies are necessary on the effect of folate in different species, also taking cleft lip and CP into consideration. PMID- 11420021 TI - Do oral flora colonize the nasal floor of patients with oronasal fistulae? AB - OBJECTIVE: To determine if oral bacteria colonize the cleft nasal floor in patients with unilateral oronasal fistula when compared with the unaffected nasal floor and whether the results obtained would be of benefit in assessing oronasal fistulae in the clinic. DESIGN: Prospective study of 26 patients with cleft palate and unilateral oronasal fistula. Microbiological culture swabs were taken from the mouth and nasal floors of patients. The unaffected nasal floor was used as a control. Bacterial isolates were identified and compared in the laboratory by a senior microbiologist. MAIN OUTCOMES MEASURE: A significant growth of oral bacteria from the cleft nasal floor when compared with the unaffected nasal floor. RESULTS: Four patients were excluded because no growth was found on any culture plate. In the remaining 22 cases, a light growth of oral flora was found in the cleft nasal floor in only 3 patients. No statistical correlation between culture of oral bacteria and the cleft nasal floor could be found (p =.12). CONCLUSIONS: The relative lack of colonization of the cleft nasal floor by oral bacteria may reflect poor transmission of bacteria through the fistula, competition with commensal nasal flora, or an inability of oral bacteria to survive in a saliva-depleted area. The investigation is not helpful in the assessment of oronasal fistulae in the clinic. PMID- 11420022 TI - Intracranial migration of halo fixation pins: a complication of using an extraoral distraction device. AB - OBJECTIVE: Distraction osteogenesis is a well-accepted technique in the treatment of patients with hypoplastic craniofacial components. Complications of distraction osteogenesis are well described in the literature. We describe a complication of using an external distraction device in a 9-year-old girl with Pfeiffer. INTERVENTION AND RESULTS: A modified Lefort III osteotomy was performed for maxillary hypoplasia with application of an external distraction halo device by a pediatric neurosurgeon. A postoperative computed tomography (CT) scan showed 0.5-cm skull penetration of the cranial pins. The pins were repositioned and the patient was followed up on a regular basis until discharge from the hospital. At 3-week follow-up, a CT scan of the head showed migration of the pins 1.5 cm intracranially. The halo was removed and repositioned at a different site. No detectable neurological sequelae from the pin penetration were noted. The patient developed cellulitis at the site of the penetration and was admitted to the hospital for a course of intravenous antibiotics. There were no other complications, and the rest of her treatment course proceeded as planned. A review of the literature on complications of halo usage as well as suggestions for their management in association with distraction osteogenesis is described. PMID- 11420023 TI - "Distraction" of grafted alveolar bone in cleft case using endosseous implant. AB - OBJECTIVE: Presentation of a specific surgical technique of vertical alveolar ridge distraction performed on an alveolar bone graft 1 year after the primary grafting procedure designed to correct a labioalveolar cleft. The graft had not obtained the results desired and presented heavily scarred mucosal tissue. TECHNIQUE: A dental implant placed within the graft once it has been entirely freed by osteotomies functions as an endless screw. To avoid vascular risk, the alveolar bone graft is detached as a whole along its total height. Consequently, no bone remains above it to provide support for a conventional alveolar distractor. An osteosynthetic miniplate, fastened in a horizontal position beneath the nasal mucosae above the graft, is used to replace the missing bony support and to stabilize the implant, which activates the distraction process. PATIENT: This study is based on our first clinical case with a follow-up of 18 months at present. RESULT: A vertical displacement of the entire graft was achieved, including its attached mucosal layer, which repositioned both bone and mucosa on a far better level in the cleft area. Thus, a normal length of abutment was obtained permitting prosthetic rehabilitation based on the same endosseous implant, which was left in place within the graft. CONCLUSION: This technique may prove particularly helpful in certain cases in which a primary alveolar bone grafting procedure has produced borderline results. In such cases, on the one hand, neither the reasonably satisfactory volume of the bone graft itself nor the poor quality of its scarred mucosal tissue argue in favor of a secondary grafting procedure. On the other hand, it is impossible to resort to currently available alveolar distracters since our choice of techniques leaves no bone support above the graft. PMID- 11420024 TI - Complete agenesis of the soft palate. AB - This report documents the physical characteristics and treatment of a patient with the extremely rare finding of complete agenesis of the soft palate. The posterior hard palate cleft and soft palate agenesis were treated with a palatal pushback procedure. PMID- 11420026 TI - Diagnosing food allergy: a test of patience. PMID- 11420025 TI - Diagnosing food allergy: a test of patience. PMID- 11420027 TI - Challenge testing in children with allergy to cow's milk proteins. AB - OBJECTIVES: To evaluate clinical response after challenge testing in infants with allergy to cow's milk proteins at diagnosis and again when these infants were aged 1 year old and had been fed an exclusion diet. MATERIAL AND METHODS: We performed a prospective study of 49 infants aged less than 6 months with a clinical history suggestive of cow's milk protein allergy, positive skin prick test and specific IgE for alpha-lactalbumin, beta-lactoglobulin and casein. In all children challenge test with cow-milk adapted formula was carried out at diagnosis. The same procedures were repeated when the children were aged 1 year but challenge testing was repeated only in children with a negative skin prick test and specific IgE antibodies to cow's milk proteins. RESULTS: At diagnosis, challenge tests produced immediate hypersensitivity reactions in 94% of infants. Late reactivity (i.e., more than 2 hours after challenge) was found in only 6% of infants, all of whom presented dyspepsia. When the infants were aged 1 year, and after results of immunological study were negative, a further challenge test was performed in 24 (49%) of lactating infants included in the study. Of these 24 infants, positive challenge was found in 5 (21%). None of the infants presented immediate symptomatology (clinical features appeared 7 days after the reintroduction of cow's milk proteins). CONCLUSIONS: Ninety-four percent of challenge tests performed at diagnosis provoked immediate reactions. The results of challenge tests after a negative skin prick test in children with normal concentrations of specific IgE were positive in 21% infants, who presented late reactivity (a mean of 7 days after milk ingestion). PMID- 11420028 TI - The comparison of the efficacy of fluticasone propionate with cetirizine in perennial allergic rhinitis. AB - BACKGROUND: Allergic rhinitis is an IgE mediated hypersensitivity reaction of the nasal mucosa characterised by nasal discharge, obstruction, and pruritus. PATIENTS AND METHODS: In this study, 43 patients with perenneal allergic rhinitis were enrolled in order to compare the efficacy of Fluticasone Propionate (FP), a corticosteroid nasal spray, with Cetirizine, a systemic oral antihistaminic preparation, which is supposed to have nonsteroidal antiinflammatory activity. Cetirizine (10 mg daily as a single dose) was administered to 22 patient for 45 days. On the other hand, FP (400 micrograms/day) was administered into each nostril twice a day in the remaining 21 patients for 45 days. Skin test was obtained from each patient before therapy. Total eosinophil count, eosinophil count in nasal smear, electrorhinomanometric investigation, PGE2 and ratio of LTC4 to LTD4 both in the serum and in the nasal secretions were determined before and after therapy. In addition, percentage of eosinophils, and mast cells count in the biopsy specimens taken from anterior edge of middle choncha were evaluated before and after therapy, and than the results were graded for each patients. RESULTS: When we compared the eosinophil count in nasal smear, eosinophil percentage and total eosinophil parameters between two groups, it was shown that FP was more effective than Cetirizine. On the other hand, when we compared the ratio of LTC4 to LTD4 in serum and nasal smear, level of PGE2 and mast cell and nasal airway resistance measured by ERM, there were non statistical difference between two groups. CONCLUSION: These results suggest that FP and Cetirizine may be used alternatively in case of an adverse reaction to any of them. PMID- 11420029 TI - Role of Zafirlukast on skin prick test. AB - BACKGROUND: Some anti-allergic and anti-asthmatic drugs should be discontinued before skin prick test. But there is no knowledge about zafirlukast effects on skin prick test. We investigate the effects of zafirlukast on cutaneous response to histamine and specific allergens. MATERIAL AND METHODS: 9 patients suffering from allergic mild asthma or allergic rhinitis and 8 healthy individuals were recruited to the study. All of them took 20 mg zafirlukast twice daily for at least 5 days. Skin prick test was performed before and after treatment by histamine and specific allergens. There was no significant difference between pre and post-treatment skin prick tests in spite of zafirlukast treatment. CONCLUSION: Zafirlukast does not affect skin reactions against histamine and specific allergens. Diagnostic skin prick test can be performed under zafirlukast treatment. PMID- 11420030 TI - Anaphylaxis after ingestion of sharon fruit. AB - BACKGROUND: The sharon fruit is the edible fruit of the persimmon tree (Diospyros kaki) which belongs to the Ebenaceae family. There are few references of allergic reaction to this fruit. We introduce a case of a 33-years old man with a anaphylactic reaction immediately after ingestion of sharon fruit. METHODS: Skin prick test (SPT) were performed by prick by prick with sharon fruit, legumes, vegetables, and fresh fruit. The test was considered positive when the average diameter of the wheal was > 3 mm. Serum-specific IgE was identified with use of the EIA, and SDS-PAGE Immunoblotting. RESULTS: The skin by prick test with sharon fruit was positive in our case, and negative in five controls patients. Specific IgE in patient serum, assayed by EIA was positive for sharon fruit, but the results of immunoblotting were non-specific. CONCLUSION: We have introduced a cause of anaphylaxis by sharon fruit that suggest an IgE mediated hypersensitivity reaction. PMID- 11420031 TI - Allergy to egg proteins. Food Allergy Committee of the Spanish Society of Pediatric Clinical Immunology and Allergy. PMID- 11420032 TI - Allergy to egg proteins. PMID- 11420035 TI - Floating the raft hypothesis: lipid rafts play a role in immune cell activation. PMID- 11420036 TI - Interferons alpha and beta as immune regulators--a new look. PMID- 11420037 TI - The TCR triggering puzzle. PMID- 11420038 TI - Involvement of SHP-1 tyrosine phosphatase in TCR-mediated signaling pathways in lipid rafts. AB - To elucidate the process of TCR-mediated signaling pathways in lipid rafts, we constructed a chimeric molecule that localizes activated SHP-1 to rafts. Raft targeting of activated SHP-1 in Jurkat-derived transfectants completely inhibited the expression of CD69 and transcriptional factors after TCR cross-linking. Whereas the inducible tyrosine phosphorylation of TCR zeta and ZAP-70 and the kinase activity of Lck were intact, phosphorylated LAT was rapidly dephosphorylated by raft targeting of activated SHP-1, leading to defects in LAT activation and subsequent downstream signaling events. Intriguingly, recruitment of endogenous SHP-1 to rafts and its association with LAT were dramatically increased after TCR engagement, suggesting that SHP-1 is involved in raft mediated T cell activation. PMID- 11420039 TI - alpha beta T cell receptor ligand-specific oligomerization revisited. AB - The mechanism of T cell receptor signaling is unclear. Included among models for TCR signaling is ligand-induced oligomerization in a fashion analogous to other cell surface receptors. Published kinetic, saturation binding, and light scattering experiments have been interpreted to suggest a propensity for soluble alpha beta TCR/peptide/MHC ectodomain complexes to oligomerize. Upon performing these experiments with soluble ectodomains of human class I and class II restricted alpha beta TCRs, we find no evidence for dimerization or oligomerization of complexes. Apparently, oligomerization in solution to a detectable extent is not a general property of soluble alpha beta TCRs or their complexes with ligand. Our results suggest that membrane-anchored, fully assembled TCRs should be studied to determine the role oligomerization plays in T cell signaling. PMID- 11420040 TI - Fc gamma RIII mediates neutrophil recruitment to immune complexes. a mechanism for neutrophil accumulation in immune-mediated inflammation. AB - Neutrophil accumulation is a hallmark of immune complex-mediated inflammatory disorders. Current models of neutrophil recruitment envision the capture of circulating neutrophils by activated endothelial cells. We now demonstrate that immobilized immune complexes alone support the rapid attachment of neutrophils, under physiologic flow conditions. Initial cell tethering requires the low affinity Fc gamma receptor IIIB (Fc gamma RIIIB), and the beta(2) integrins are additionally required for the subsequent shear-resistant adhesion. The attachment function of Fc gamma RIIIB may be facilitated by its observed presentation on neutrophil microvilli. In vivo, in a model of acute antiglomerular basement membrane nephritis in which immune complexes are accessible to circulating neutrophils, Fc gamma RIII-deficient mice had a significant reduction in neutrophil recruitment. Thus, the interaction of immune complexes with Fc gamma RIII may mediate early neutrophil recruitment in immune complex-mediated inflammation. PMID- 11420041 TI - Il-6 and its soluble receptor orchestrate a temporal switch in the pattern of leukocyte recruitment seen during acute inflammation. AB - During acute inflammation, leukocyte recruitment is characterized by an initial infiltration of neutrophils, which are later replaced by a more sustained population of mononuclear cells. Based on both clinical and experimental evidence, we present a role for IL-6 and its soluble receptor (sIL-6R) in controlling this pattern of leukocyte recruitment during peritoneal inflammation. Liberation of sIL-6R from the initial neutrophil infiltrate acts as a regulator of CXC and CC chemokine expression, which contributes to a suppression of neutrophil recruitment and the concurrent attraction of mononuclear leukocytes. Soluble IL-6R-mediated signaling is therefore an important intermediary in the resolution of inflammation and supports transition between the early predominantly neutrophilic stage of an infection and the more sustained mononuclear cell influx. PMID- 11420042 TI - TGF-beta released by apoptotic T cells contributes to an immunosuppressive milieu. AB - T cell apoptosis is critical to development and homeostasis of the immune system. The most salient feature of apoptosis is the lack of an attendant inflammatory response or tissue damage. Here, we present evidence that apoptotic T cells release TGF-beta, thereby contributing to an immunosuppressive milieu. Apoptotic T cells released not only latent but also bio-active TGF-beta. Nonetheless, TGF beta transcription was not upregulated, suggesting release of existing rather than synthesis of new TGF-beta. Localized within the intracellular membrane-bound compartment, which includes mitochondria, TGF-beta was redistributed into the cytosol following loss of mitochondrial membrane potential. TGF-beta secreted from apoptotic T cells inhibited proinflammatory cytokine production by activated macrophages to foster immune suppression. These findings broaden the potential mechanisms whereby induction of immune tolerance or deficiency occurs through T cell deletion. PMID- 11420043 TI - An essential role for Daxx in the inhibition of B lymphopoiesis by type I interferons. AB - Interferon-alpha and -beta inhibit the interleukin-7-mediated growth and survival of T and B lymphoid progenitors via an unknown, STAT1-independent pathway. Gene expression profile analysis of interferon-beta-treated progenitor B cells revealed enhanced Daxx expression, with concomitant Daxx protein increase and nuclear body translocation. The interferon effects included downregulation of cell cycle regulating genes and cell cycle arrest, followed by Bcl-2 downregulation and apoptosis. Daxx antisense oligonucleotides rescued the interferon-treated pro-B cells from growth arrest and apoptosis in parallel with the reduction of nuclear Daxx. These findings implicate the gene repressor function of Daxx in interferon-induced apoptosis of lymphoid progenitors. PMID- 11420044 TI - MHC class II expression is regulated in dendritic cells independently of invariant chain degradation. AB - We have investigated the mechanisms that control MHC class II (MHC II) expression in immature and activated dendritic cells (DC) grown from spleen and bone marrow precursors. Degradation of the MHC II chaperone invariant chain (Ii), acquisition of peptide cargo by MHC II, and delivery of MHC II-peptide complexes to the cell surface proceeded similarly in both immature and activated DC. However, immature DC reendocytosed and then degraded the MHC II-peptide complexes much faster than the activated DC. MHC II expression in DC is therefore not controlled by the activity of the protease(s) that degrade Ii, but by the rate of endocytosis of peptide-loaded MHC II. Late after activation, DC downregulated MHC II synthesis both in vitro and in vivo. PMID- 11420045 TI - Adenovirus L4-100K assembly protein is a granzyme B substrate that potently inhibits granzyme B-mediated cell death. AB - Cytotoxic lymphocytes kill virus-infected target cells and play a critical role in host recovery from viral infections. Granzyme B (GrB) is a cytotoxic lymphocyte granule protease that plays a critical role in mediating cytotoxicity. In these studies, we demonstrate that the adenovirus assembly protein L4--100K (100K) is a GrB substrate that prevents cytotoxic lymphocyte granule-induced apoptosis in infected target cells by potently inhibiting GrB. This inhibition is absolutely dependent on Asp-48 in 100K, found within a classic GrB consensus motif. 100K is the first viral protein described that exclusively targets the GrB pathway. It represents a novel class of viral protease inhibitor, in which an essential, multifunctional viral protein, which is vulnerable to specific proteolysis by GrB, expresses inhibitory function against that protease. PMID- 11420046 TI - Nef triggers a transcriptional program in T cells imitating single-signal T cell activation and inducing HIV virulence mediators. AB - Gene expression profiling was used to explore the role of Nef in HIV. Nef induces a transcriptional program in T cells that is 97% identical to that of anti-CD3 T cell activation. This program is inhibited in the presence of cyclosporin. A requirement for TCR zeta and ZAP-70 is demonstrated for formation of the complete profile. Among eight factors particular to the anti-CD3 activation profile are IL16 and YY1, negative regulators of HIV transcription. In contrast, Nef exclusively upregulates factors positively regulating HIV, including Tat-SF1, U1 SNRNP, and IRF-2. New genes associated with Nef include CDK9, the induction of which enhances Tat function. Thus, Nef acts as a master switch early in the viral life cycle, forcing an environment conducive to dynamic viral production. PMID- 11420047 TI - Pax5/BSAP maintains the identity of B cells in late B lymphopoiesis. AB - The B lineage commitment factor Pax5 (BSAP) is expressed throughout B cell development. To investigate its late function, we generated a mouse strain carrying a floxed Pax5 allele that was conditionally inactivated by CD19-cre or Mx-cre expression. Pax5 deletion resulted in the preferential loss of mature B cells, inefficient lymphoblast formation, and reduced serum IgG levels. Mature B cells radically changed their gene expression pattern in response to Pax5 inactivation. Most B cell antigens were downregulated on the cell surface, and the transcription of B cell-specific genes was decreased, whereas the expression of non-B lymphoid genes was activated in Pax5-deficient B cells. These data demonstrate that Pax5 is essential for maintaining the identity and function of B cells during late B lymphopoiesis. PMID- 11420048 TI - Regulation of mast cell survival by IgE. AB - Mast cells play critical roles in hypersensitivity and in defense against certain parasites. We provide evidence that mouse mast cell survival and growth are promoted by monomeric IgE binding to its high-affinity receptor, Fc epsilon RI. Monomeric IgE does not promote DNA synthesis but suppresses the apoptosis induced by growth factor deprivation. This antiapoptotic effect occurs in parallel with IgE-induced increases in Fc epsilon RI surface expression but requires the continuous presence of IgE. This process does not involve the FasL/Fas death pathway or several Bcl-2 family proteins and induces a distinctly different signal than Fc epsilon RI cross-linking. The ability of IgE to enhance mast cell survival and Fc epsilon RI expression may contribute to amplified allergic reactions. PMID- 11420049 TI - Monomeric IgE stimulates signaling pathways in mast cells that lead to cytokine production and cell survival. AB - Although IgE binding to mast cells is thought to be a passive presensitization step, we demonstrate herein that monomeric IgE (mIgE) in the absence of antigen (Ag) stimulates multiple phosphorylation events in normal murine bone marrow derived mast cells (BMMCs). While mIgE does not induce degranulation or leukotriene synthesis, it leads to a more potent production of cytokines than IgE + Ag. Moreover, mIgE prevents the apoptosis of cytokine-deprived BMMCs, likely by maintaining Bcl-X(L) levels and producing autocrine-acting cytokines. The addition of Ag does not increase this IgE-induced survival. Since IgE concentrations as low as 0.1 microg/ml enhance BMMC survival, elevated plasma IgE levels in humans with atopic disorders may contribute to the elevated mast cell numbers seen in these individuals. PMID- 11420050 TI - The effect of short-term tooth intrusion on human pulpal blood flow measured by laser Doppler flowmetry. AB - The purpose was to examine the effect of brief intrusive forces on human pulpal blood flow (PBF). Laser Doppler flowmetric measurements were made from 17 vital upper left central incisors of 17 participants who had clinically healthy tooth crowns and periodontal tissues. Brief intrusive forces (0.5,1,5 N; duration 20 s) were applied to the incisal edges of the examined teeth, and apical displacement of the teeth and the PBF were measured simultaneously. Recordings were made with and without an opaque rubber dam applied to the examined teeth. Intrusive force significantly reduced PBF flux both with and without the dam (P<0.05, Friedman analysis). The results indicate that transient apical displacement can reduce PBF. PMID- 11420051 TI - Ca2+-independent phospholipase A2 activity in apical plasma membranes from the rat parotid gland. AB - An apical-enriched plasma membrane fraction (A-PM) was prepared from rat parotid gland by Mn2+ precipitation. In this fraction, phosphatidylcholine (PC) labelled at the sn-2 position was mainly decomposed into two labelled compounds (free fatty acid and 1,2-diacylglycerol) under Ca2+-free conditions. Studies using double-labelled PC and 2,3-diphosphoglycerate (as a phospholipase D inhibitor) showed that they were produced through different pathways: free fatty acid was released by phospholipase A2 (PLA2) while 1,2-diacylglycerol may be produced by sequential action of phospholipase D and phosphatidate phosphatase. The PLA2 in A PM did not require Ca2+ for its activity and was highly activated by Triton X-100 and ATP. The inhibitor of the well-documented Ca2+-independent PLA2, bromoenol lactone, did not inhibit the PLA2 activity in A-PM. Although PLA2 activity was detected in other subcellular fractions, the highest specific activity was in A PM. Its distribution among various fractions was roughly similar to that of the marker enzyme of apical plasma membranes. These findings suggested that Ca2+ independent PLA2 activity is present in apical plasma membranes from rat parotid gland. In addition, to clarify the involvement of the PLA2 in exocytosis, the fusion of exogenous PLA2-treated membranes with secretory granules was examined by fluorescence dequenching assay. This study clearly demonstrated the facilitation of fusion by PLA2 treatment, which suggests some involvement of apical PLA2 in saliva secretion. PMID- 11420052 TI - Identification and regulation of K+ and Cl- channels in human parotid acinar cells. AB - The properties of K+ channels in these cells were studied using patch-clamp methods. Two channels, with conductances of 165+/-13 pS (n=6) and 30+/-1 pS (n=3), were identified in single-channel experiments. In cell-attached patches the reversal potentials were -67+/-8 and -74+/-2 mV for the large and small conductance channel, respectively, suggesting that both channels are K+ selective. The large conductance channel was also shown to be K+-selective in inside-out patches. The open probability (P(o)) of this channel was increased at depolarizing potentials and by increasing intracellular Ca2+ concentration ([Ca2+]i). These properties suggest that the large conductance channel is a 'maxi' Ca2+-activated K+ channel (BK(Ca)). The small conductance channel was not observed in inside-out patches. Carbachol (CCh; 10(-5) M) activated the BK(Ca) channel, but not the small conductance channel, in cell-attached patches. CCh also caused a dose-dependent increase in [Ca2+]i measured by fura-2 in microspectrofluorimetric studies, with a half-maximal response at approximately 3x10(-6) M. Neither isoproterenol (10(-5) M) nor substance P (10(-6) M) affected K+-channel activity or [Ca2+]i. In whole-cell experiments, CCh caused an increase in outward current. Charybdotoxin (10(-7) M), a BK(Ca) blocker, inhibited a large component of the CCh-induced current. A large component of the charybdotoxin insensitive current may be carried by Ca2+-activated Cl- channels, which were also observed in human parotid acinar cells. The results indicate that BK(Ca) channels make a significant contribution to the whole-cell conductance in human parotid acinar cells. PMID- 11420053 TI - A longitudinal electromyographic study of the postnatal maturation of mastication in the rabbit. AB - At 2 weeks of age, infant rabbits show chewing movements that resemble those of the adult animal. Previous studies have shown that, at that stage, the accompanying masticatory motor pattern is clearly similar to the suckling motor pattern. As early as 4 weeks, chewing muscle activity is indistinguishable from the adult chewing motor pattern. These reports suggest that the adult chewing motor pattern is developed from the suckling motor pattern. In this study, the chewing motor pattern in the intermediate period (between 2 and 4 weeks of age) was investigated by means of fine-wire electromyography and jaw tracking. Maturation of masticatory movements was found to have two phases. Maximum gape increased in the first few days and was followed by strong development of transverse jaw excursions after the age of 17 days. The increase in jaw excursions was brought about by changes in motor behaviour and facilitated by the development of smooth occlusal surfaces. The changes in motor behaviour were: (1) the level of activity of the balancing-side muscles became more equal to that of the working side; (2) the timing of digastric muscle activity became asymmetrical at the age of 17 days; (3) the peak activity of masseter, temporalis, medial pterygoid and lateral pterygoid muscle portions was gradually shifted or prolonged into the power-stroke phase. It can be concluded that the masticatory contraction pattern shifts from one derived from the suckling contraction pattern at the age of 14 days to one almost similar to the adult chewing pattern at the age of 23 days. PMID- 11420054 TI - Differences in myosin heavy-chain composition between human jaw-closing muscles and supra- and infrahyoid muscles. AB - Jaw-closing muscles have architectural features suited to force production; supra and infrahyoid muscles are better adapted to produce velocity and displacement. It was hypothesized that this difference in function would be reflected in myosin heavy-chain (MyHC) composition (equivalent to contraction velocity) and fibre type cross-sectional area (equivalent to force). MyHC composition was determined in muscles obtained from eight human cadavers, using monoclonal antibodies against MyHC isoforms. Jaw closers contained 4.2 times fewer type IIA fibres and 5.2 times more hybrid fibres than suprahyoid muscles, and 3.9 times fewer type IIA fibres and 3.2 times more hybrid fibres than the infrahyoid muscles. In the jaw closers, MyHC-I was expressed in approx. 70% of all fibres (pure+hybrid), in the suprahyoid muscles in approx. 40%, and in the infrahyoid muscles in approx. 46%. In the jaw closers, type I fibres were 40% larger in diameter than in the supra- and infrahyoid muscles. It can be concluded that the jaw closers have characteristics of slow muscles, and that the supra-/infrahyoid muscles have characteristics of fast muscles. PMID- 11420055 TI - Alteration of odontoblast osteonectin expression following dental cavity preparation. AB - Cavity preparation can increase the active synthesis and secretion of non collagenous proteins by odontoblasts, thus resulting in the deposition of tertiary dentine. In this study, the effect of cavity preparation on osteonectin expression was examined in odontoblasts of the rat tooth pulp. A class V cavity was prepared in rat first molars to stimulate odontoblastic secretory activity, and the animals were killed at various intervals. In the normal pulp, osteonectin immunoreactivity was detected in odontoblasts but not other cells. At 1 day after cavity preparation, immunoreactivity had diminished beneath the cavity. At 3 days, strong immunoreactivity could be detected in odontoblasts beneath the cavity. Numerous round cells underlying the odontoblastic layer also demonstrated immunoreactivity. Thereafter, the intensity of osteonectin immunoreactivity in odontoblasts beneath tertiary dentine decreased gradually, and at 30 and 60 days, it was weaker than in normal pulp. These findings suggest that osteonectin is actively synthesized by odontoblasts underlying a cavity in the initial stage of tertiary dentine formation. PMID- 11420056 TI - Tissue specificity of lymphocyte migration into sheep gingival tissue. AB - T cells show a bias in their migration pathways: some migrate preferentially to peripheral lymph nodes, some to mucosal tissues and some to peripheral tissues such as skin. The aim here was to determine the types of T cells that migrate preferentially into inflamed gingival tissue and compare this migration to that found in inflamed subcutaneous and mucosal tissues. The experiments were designed so that the simultaneous 3 h localization of two, differentially radiolabelled, lymphocyte populations (subcutaneously and mucosally derived) into sites of purified protein derivative/bacillus Calmette-Guerin-induced, delayed-type hypersensitivity, inflammatory lesions in skin, bowel and gingiva in the sheep model could be compared. The relative migration of two populations in each of the tissues was expressed as a ratio of the radioactivity of intestinal/subcutaneous lymphocytes recovered from that tissue. From nine experiments, the ratios [mean+/ S.E.M. (n)] for skin, bowel and gingiva were 0.53+/-0.02 (84), 1.98+/-0.11 (85), and 0.73+/-0.05 (29), respectively. These findings suggest that inflammation in skin and gingiva favoured the localization of subcutaneously derived lymphocytes (ratio significantly <1, P<0.025), while in bowel, the localization of intestinally derived lymphocytes was favoured (ratio significantly >1, P<0.025). Statistical analysis demonstrated that the relative localization of the two lymphocyte populations to the gingival lesions differed significantly from that for inflamed skin and bowel lesions (P<0.05). When tumour necrosis factor-alpha was used as a non-antigenic inflammatory agent to induce lymphocyte migration into skin and gingiva, a similarly greater increase in the localization of subcutaneously derived lymphocytes was detected, but the relative localization of lymphocytes was not significantly different between the two tissues. Therefore, it appears that there is tissue specificity in the migration of lymphocytes into the inflamed gingival tissues and that antigen is required for distinct tissue specific lymphocyte traffic to occur. PMID- 11420057 TI - The effects of human mast-cell products and of phenytoin on androgen 5alpha reductase expression in human gingival fibroblasts. AB - The aim was to elucidate mechanisms for phenytoin-induced gingival overgrowth, using effects of type 1 human mast cell (HMC-1) supernatant and histamine on the expression of 5alpha-reductase in human gingival fibroblasts (HGF), and the effects of phenytoin on this activity. Duplicate incubations of HGF in Eagle's minimum essential medium (MEM) were performed with [14C] testosterone and serial concentrations of mast-cell histamine (1-100 microgram/ml)/HMC-1 culture supernatant at serial dilutions of 1-100 microl/ml for 24 h. Other experiments employed optimal concentrations of HMC-1 supernatant (10 microl/ml) or histamine (8 microgram/ml) and phenytoin (1 and 5 microgram/ml), alone and in combination, using two androgen substrates, [14C] testosterone and [14C] 4-androstenedione. At the end of a 24-h incubation the medium was solvent-extracted for steroid metabolites, analysed and quantified in a radioisotope scanner. HMC-1, histamine (n=3) and phenytoin (n=6) significantly increased the synthesis of dihydrotestosterone and 4-androstenedione by up to 80% (P<0.01); the combination of HMC-1 and phenytoin caused two-fold increases (n=6; P<0.01). The incubations with histamine alone and in combination with phenytoin showed significant stimulation of dihydroxytestosterone and the diols alone and in combination, which was less pronounced in combination. This investigation demonstrates significant stimulation of 5alpha-reductase activity in human gingival fibroblasts by mast-cell supernatant and a specific product histamine, alone and in combination with phenytoin. Androgen 5alpha-reductase-mediated anabolic actions in connective tissue are well documented. The findings suggest a novel hypothesis that mast-cell mediated androgen action in the gingiva in response to phenytoin could contribute to gingival overgrowth. PMID- 11420058 TI - A study of occlusal variation in the primary dentition of Australian twins and singletons. AB - In this study, we quantified the extent of variation in different occlusal features of Australian children of European descent with complete primary dentitions, but no permanent teeth present in the mouth. The study group consisted of 412 individuals, aged between 3 and 7 years, including 70 monozygous (MZ) twin pairs, 68 same-sexed dizygous (DZ) twin pairs, 11 opposite-sexed DZ twin pairs, and 114 singletons. Occlusal traits, including inter-dental spacing, incisal overbite and overjet, arch breadth and arch depth, were obtained directly from dental casts or indirectly from photocopies or impressions of the casts. Descriptive statistics summarised the data and indicated that distributions were similar to those published for other populations. Univariate genetic analysis, using the structural equation modelling package Mx, was carried out on the quantitative data using the normal assumptions of the twin model. Genetic modelling indicated that a model incorporating additive genetic (A) and unique environmental variation (E) was the most parsimonious for interdental spacing, overbite, overjet and arch dimensions. Estimates of heritability for interdental spacing ranged from 0.62 to 0.81. Estimates for overbite and overjet were 0.53 and 0.28, respectively, and estimates for arch dimensions ranged from 0.69 to 0.89. These results indicated a moderate to relatively high genetic contribution to observed variation. PMID- 11420059 TI - The mechanism of palatal clefting in the Col11a1 mutant mouse. AB - The occurrence of cleft palate in mutant mice offers an opportunity to understand the possible role of specific genes in palatogenesis. Here, cleft palate in mice carrying the chondrodysplasia (cho) defect, which consists of an autosomal recessive mutation in the collagen gene Col11a1, was investigated. The proposed cause of cleft palate in cho homozygous mice is failure of the palatal shelves to adhere and make contact due to mandibular growth abnormalities. Another cause of cleft palate that has recently been demonstrated in other animal models is failure of the midline epithelial seam forming between the shelves to undergo epithelial-mesenchymal transformation (EMT). The present strategy to test the likelihood of this second possibility was to culture the unfused cho/cho palatal shelves at different stages of development to see if they were capable of adhering and undergoing EMT in vitro. By using carboxydichlorofluorescein succinimidyl ester to trace the fate of the medial-edge epithelium (MEE), it was shown that cho/cho palates have full potential for MEE adherence and EMT up to embryonic day 17.5/18.5, when epithelia keratinize before birth, preventing the adherence of both the normal and homozygous palatal shelves. Thus, the major effect of the mutant collagen gene on the palate is likely to be via mandibular growth disruption. The possibility that unfused palatal shelves in other clinical syndromes can adhere and undergo EMT if brought into contact at appropriate times before birth has important therapeutic implications. PMID- 11420060 TI - In vitro evaluation of different remineralization periods in improving the resistance of previously eroded bovine dentine against tooth-brushing abrasion. AB - One dentine specimen was prepared from each of 90 bovine incisors. The samples were then evenly distributed among nine groups (A-I) and submitted to 10 alternating de- and re-mineralization cycles, including abrasion by tooth brushing. Each cycle started with a demineralization using the erosive soft drink Sprite Light for 1 min, followed by storing the samples in pooled human saliva for a total of 240 min. The specimens were removed from the saliva at different intervals (group A, 0 min; B, 15 min; C, 30 min; D, 45 min; E, 60 min; F, 90 min; G, 120 min) and brushed in an automatic brushing machine. Groups H (erosion, but no brushing) and I (no erosion, but brushing), which were also stored in saliva for 240 min, served as controls. After these cycles, loss of dentine was determined by profilometry, producing the following values (mean+/-S.D.), which were analysed statistically (P< or = 0.05): group A (5.03+/-1.49 microm), B (4.44+/-1.09 microm), C (4.91+/-0.95 microm), D (5.47+/-1.52 microm), E (5.29+/ 1.45 microm), F (4.76+/-0.74 microm), G (5.16+/-0.71 microm), H (2.61+/-1.31), I (1.11+/-0.39). Groups A-G had no significant differences, but they showed a significantly greater loss of dentine than groups H and I. It is concluded that the abrasion resistance of eroded bovine dentine is still decreased after a remineralization period of 120 min. PMID- 11420061 TI - The effect of cyclosporin A on the activity of matrix metalloproteinases during the healing of rat molar extraction wounds. AB - Cyclosporin A (CyA) is a cyclic peptide used as an immunosuppressive agent because it can block the synthesis of interleukin-2 and other cytokines produced by CD4+ lymphocytes. It is widely used for the prevention of allograft rejection and treatment of autoimmune diseases. Several side-effects of CyA treatment have been reported, among which are chronic nephrotoxicity, hepatotoxicity and neurotoxicity, lymphoproliferative neoplasms, hypertension, thromboembolic complications and gingival overgrowth. Here, using a rat molar model, it is demonstrated that CyA immunosuppression inhibits the activity of matrix metalloproteinases 2 and 9 in the early phase of granulation tissue in the healing dental socket. These observations suggest that CyA may interfere with the wound healing following dental extractions. PMID- 11420062 TI - Pharmacodynamic and receptor binding changes during chronic lorazepam administration. AB - To assess pharmacodynamic and neurochemical aspects of tolerance, lorazepam (2 mg/kg/day), or vehicle was administered chronically to male Crl: CD-1(ICR)BR mice via implantable osmotic pump. Open-field behavior, benzodiazepine receptor binding in vitro, receptor autoradiography, and muscimol-stimulated chloride uptake were examined at both 1 and 14 days. Open-field activity was depressed in lorazepam-treated animals on Day 1. On Day 14, open-field parameters were indistinguishable from those of vehicle-treated animals, indicating behavioral tolerance. Benzodiazepine binding, as determined by the specific binding of [125I]diazepam, was also decreased in cortex on Day 14. Hippocampal binding was unchanged following chronic lorazepam exposure. Apparent affinity in cortical membrane preparations was unchanged, indicating that altered ligand uptake was due to decreased receptor number. Muscimol-stimulated chloride uptake into cortical synaptoneurosomes from lorazepam-treated animals was not significantly different on Day 1 or Day 14 compared to vehicle-treated animals. These results confirm that down-regulation of benzodiazepine receptor binding is closely associated with behavioral tolerance to benzodiazepines. These observed changes in binding are not necessarily associated with robust changes in receptor function. PMID- 11420063 TI - GABA(A) but not GABA(B) receptor stimulation induces antianxiety profile in rats. AB - The effect of GABA receptor agonists and antagonists on anxiety behavior in rats in the elevated-plus-maze has been investigated. The increase in two parameters of %open arm entries (%OAE) and %time spent in the open arms (%OAT) and decrease in the %time spent in closed arm (%CAT) was considered as antianxiety effects. Intracerebroventricular (i.c.v.) injection of different doses of the GABA(A) receptor agonist muscimol (0.25, 0.5, and 1 microg/rat) increased %OAE and %OAT and decreased %CAT in rats dose-dependently. The higher response was obtained with 1 microg/rat of the drug. Neither icv (0.05, 0.1, and 0.2 microg/rat) nor intraperitoneal (i.p.) (1, 2, and 4 mg/kg) injection of the GABA(B) receptor agonist baclofen altered %OAE, %OAT, and %CAT. However, the GABA(B) receptor antagonist CGP35348 (5, 10, and 30 microg/rat i.c.v.) increased %OAE and %OAT and decreased %CAT in the animals. The response induced by injection of muscimol (0.5 microg/rat i.c.v.) or administration of CGP35348 (10 microg/rat i.c.v.) was reduced by i.c.v. (1, 2, and 4 microg/rat) or i.p. (0.25, 0.5, and 0.75 mg/kg) injection of the GABA(A) receptor antagonist bicuculline, except the effect of CGP35348 on %CAT which was not significantly altered by i.p. administration of bicuculline. Ip but not i.c.v. administration of bicuculline by itself reduced both %OAE and %OAT but did not alter %CAT. None of the drugs altered the locomotor activity of the animals. The current findings support our hypothesis that the anxiolytic effects of GABA(B) antagonist are mediated by autoreceptor blockade-induced release of endogenous GABA, which in turn activates postsynaptic GABA(A) receptors. PMID- 11420064 TI - Involvement of protein kinase C and tyrosine kinase in lipopolysaccharide-induced anorexia. AB - Injections of lipopolysaccharide (LPS, 3 microg) into the lateral ventricle elicited anorexia with fever and also decreased body weight in rats. The LPS induced anorexia was inhibited by intracerebroventicular (i.c.v.) injections of anti-interleukin (IL)-1beta antibody (Ab), chelerythrine, genistein and tyrphostin 46, but not by injections of indomethacin. Consecutive injections of orthovanadate and LPS (0.3 microg, a dose of LPS that did not show any effect on food intake, body weight or body temperature) reduced body weight, but did not induce anorexia. On the other hand, injections of IL-1beta (50 ng) did not influence food intake, although they decreased body weight and produced fever. The IL-1beta-induced decrease in body weight was inhibited by injections of genistein, but not by injections of chelerythrine or indomethacin. These findings suggest that the LPS-induced anorexia is independent of hyperthermia and involves IL-1beta generation, tyrosine kinase (TK) and protein kinase C (PKC). This is the first in vivo evidence that activation of TK and PKC induced by LPS is linked to anorexia. PMID- 11420065 TI - The effects of pyridostigmine bromide, permethrin, and DEET alone, or in combination, on fixed-ratio and fixed-interval behavior in male and female rats. AB - Concurrent exposure to pyridostigmine bromide (PB), permethrin (PERM) and/or N,N diethyl-m-toluamide (DEET) may have contributed to the development of a syndrome that appears to have afflicted military personnel who served during the Gulf War. The present experiment sought to evaluate the behavioral effects of these compounds alone, or in various combinations, in male and female rats. Subjects were exposed to a multiple fixed-ratio (FR) 50, fixed-interval (FI) 2-min schedule of reinforcement. PB dose-dependently decreased FR and FI response rates. FR responding was disrupted by lower doses and there were no differences between the sexes. PERM vehicle administration decreased response rates maintained by both schedules of reinforcement; this was offset by an increase in response rate after the administration of the intermediate dose of PERM. The highest dose of PERM decreased both FR and FI response rates. FR rates in male rats were more disrupted than those in female rats. Only the highest dose of DEET decreased FR and FI response rates in male and female rats. FR rates were more disrupted in female rats than in male rats. Synergistic effects were only observed when FI response rates decreased in male rats upon exposure to half the low dose of PB with half the low dose of PERM or half the low dose of PB with half the low dose of DEET. The results of this experiment thus show that small doses of PB, PERM and DEET disrupt well-established, schedule-controlled behavior in male and female rats in a schedule- and gender-dependent manner; schedule dependent and gender-dependent synergistic effects were also observed. The mechanism by which the compounds exert these behavioral effects remains to be determined. PMID- 11420066 TI - Strain differences in the anxiolytic effects of losartan in the mouse. AB - Anxiolytic effects of the angiotensin AT(1) receptor antagonist losartan were studied in the elevated plus maze (EPM) and the light/dark test (LDT) in different mouse strains as were responses to angiotensin II and acetylcholine in isolated ascending colon. There were no significant strain differences in behaviour on the EPM, and diazepam was anxiolytic in C57BL/6, DBA/2 and BKW mice. Losartan was anxiolytic in BKW only. In the LDT, there were significant strain differences, with BKW mice exhibiting greatest anxiety-like behaviour; losartan was ineffective in this test. In vitro responses to angiotensin II and acetylcholine were significantly smaller in BKW than in C57BL/6 and DBA/2. These results indicate that the mouse strain exhibiting least angiotensin receptor function is the most responsive to the anxiolytic effects, suggesting a possible relationship between angiotensin receptor function and anxiolytic response to losartan. PMID- 11420067 TI - Immediate and delayed voluntary ethanol effects on motor performance, learning and inhibition in rats. AB - The effects of prolonged voluntary ethanol consumption on psychomotor performance, operant conditioning and inhibition were examined in adult male Wistar rats. Animals were food deprived and alcohol or control solution was available 1 h/day during 15 days, with free water for the rest of the day. Then, rats were tested in a two-bottle paradigm (solution and water available) for 1 h/day during 19 days, and subjects were tested daily for psychomotor performance and operant conditioning immediately or 6 h after (delayed) the solution access. Psychomotor performance was tested in an 80 degrees -inclined screen. Successive conditioning phases were: free shaping (FS), continuous reinforcement (CRF), operant extinction (EXT), successive discrimination (DIS) and two-stimuli test (TST). Alcohol consumption deteriorated psychomotor performance and improved the animal's ability to learn simple associations between stimuli and responses (free shaping and extinction), in immediate and delayed groups. Finally, alcohol deteriorated behavioral inhibition (DIS and TST) tested immediately after drinking. Taken together, results suggest that prolonged voluntary ethanol intake could induce permanent psychomotor impairment and associative learning facilitation, and also an impairment of the inhibition related to the intoxication state. PMID- 11420068 TI - Chronic morphine consumption decreases wheel running and wheel running-reinforced behavior in rats. AB - The purpose of this experiment was to evaluate the effects of morphine self administration on wheel running and wheel running-reinforced lever pressing in rats. The home cage was equipped with a bottle that contained either water, a saccharin-flavored 0.5-mg/ml morphine solution, or saccharin (0.25%). The bottle was available for either 1 or 3 h. The bottle was then removed, and 20-22 h after removal, the rats were moved to an operant chamber in which lever presses earned 15 s access to a running wheel (according to a variable interval (VI) 40-s schedule). The morphine condition was in effect for 69 days, and consumption gradually increased to a level of 67 mg/kg/day. During the morphine condition, wheel running and lever pressing decreased. Following the removal of morphine, (so that the home-cage bottles provided a 0.25% saccharin solution), the two instrumental behaviors increased to the pre-morphine (water) levels. However, the increases were not immediate, and in the first post-morphine session, lever pressing and wheel turning remained at the depressed morphine level. The post morphine increase in lever pressing was substantially larger than the increase in wheel running. The results support the hypothesis that chronic opiate consumption reduces the frequency of some nondrug-related behaviors, and that this, in turn, increases preference for the opiate. PMID- 11420069 TI - The role of dopamine in the locomotor stimulant effects and tolerance to these effects of caffeine. AB - Current evidence indicates that the acute locomotor stimulant effects of caffeine involve dopamine (DA) receptor activation; however, few studies have investigated the role of DA receptors in mediating the development of tolerance to caffeine. Therefore, the present study was designed to determine the degree to which DA receptors mediate the development of tolerance to the locomotor stimulant effects of caffeine. Caffeine was examined alone and in combination with haloperidol (HAL), GBR 12909, nisoxetine and fluoxetine. HAL dose-dependently and completely blocked the acute effects of caffeine on locomotor activity, and the highest dose of GBR 12909 enhanced the effects of caffeine. Neither nisoxetine nor fluoxetine altered the effects of caffeine. HAL was infused via osmotic pumps (0.1 mg/kg/day) during a 14-day regimen of chronic caffeine administered in a caffeinated drinking solution ( approximately 136 mg/kg/day). HAL did not block the development of tolerance to the locomotor stimulant effects of caffeine, but did impair the recovery from tolerance following withdrawal of caffeine. [3H]SCH 23390 (DA D(1)) binding sites were downregulated in the nucleus accumbens and striatum and were upregulated in the prefrontal cortex of caffeine-treated vs. control rats; however, the affinity of [3H]SCH 23390 for these binding sites was unaltered. There were no differences between the caffeine-treated and control rats in number or affinity of [3H]spiperone (DA D(2)) binding sites. These results suggest that, although HAL did not alter the development of tolerance to caffeine, changes in DA D(1) receptors could be one component of the mechanism underlying caffeine-induced tolerance. PMID- 11420070 TI - Embryonic cocaine exposure and corticosterone: serotonin(2) receptor mediation. AB - Cocaine activates the mature hypothalamic-pituitary-adrenal (HPA) axis, increasing corticosterone concentrations in animals and humans and serotonin(2) receptors (5-HT(2)) are involved in this effect. Although prenatal cocaine exposure is associated with altered responsiveness of the HPA axis to "stress" and serotonergic compounds postnatally, it is unknown whether cocaine directly activates the embryonic HPA axis or if 5-HT(2) receptors are involved. Domestic chicken eggs with viable embryos were exposed to either the 5-HT(2) receptor agonist dimethoxyiodophenylaminopropane (DOI: 0.4, 0.8, or 1.2 mg/kg egg) or saline on embryonic day 18 (E18). In a second study, the 5-HT(2) antagonist ritanserin (0.3 mg/kg egg, a dose found effective against other effects of DOI or cocaine) or vehicle was administered on E17, prior to treatment on E18 with either saline or cocaine (5 injections of 12 mg/kg egg, equivalent to a total dose of 3.5 mg/egg). Radioimmunoassay was used to measure serum corticosterone from blood samples taken approximately 1-2 h after drug injections. DOI significantly raised corticosterone in a dose-related fashion. Cocaine-induced corticosterone elevations were blocked by pretreatment with ritanserin, whereas ritanserin by itself did not affect corticosterone concentrations. These data indicate that 5-HT(2) receptors are involved in cocaine's effect on the HPA axis during late chicken embryogenesis. PMID- 11420071 TI - The inability of CCK to block (or CCK antagonists to substitute for) the stimulus effects of chlordiazepoxide. AB - To further examine the relationship between cholecystokinin (CCK) and GABA, the present study assessed the ability of the CCK-A antagonist devazepide and the CCK B antagonist L-365,260 to substitute for the stimulus effects of chlordiazepoxide (CDP), as well as the ability of CCK-8s to block these effects, in female Long Evans rats within the conditioned taste aversion baseline of drug discrimination learning. Both devazepide and L-365,260 failed to substitute for the discriminative stimulus properties of CDP, and CCK-8s failed to block its stimulus effects. The benzodiazepine diazepam did substitute for, and the benzodiazepine antagonist flumazenil did block, the stimulus effects of CDP. This suggests that the lack of substitution for, or antagonism of, CDP by the CCK antagonists and CCK-8s, respectively, was not due to the inability of the present design to assess such effects. Possible bases for the current findings, e.g., necessity of an anxiogenic baseline, drug and receptor specificity, as well as the dose-response nature of the interaction, were discussed. Given that a relationship between CCK and GABA has been reported in other designs, the present results suggest that such a relationship may be preparation specific. PMID- 11420072 TI - Dose-sensitive excitation and inhibition of spontaneous amygdala activity by propranolol. AB - The effect of systemically administered propranolol was determined on spontaneous activity of neurons in the central nucleus (CeA) of the amygdala, a brain site implicated in fear-related learning and memory. Extracellular recordings of single units in the CeA were obtained in vivo from rats administered saline or the centrally and peripherally acting beta-adrenergic receptor blocker propranolol (4, 7, 10 mg/kg i.p.). The high dose (10 mg/kg) of propranolol markedly increased spontaneous activity of CeA neurons. In contrast, the low (4 mg/kg) and intermediate (7 mg/kg) doses of propranolol significantly decreased spontaneous CeA activity, with the suppressant effect of propranolol on CeA firing rates weakening as the dosage increased from 4 to 7 mg/kg. These results suggest that (1) spontaneous activity of CeA neurons is tonically influenced by competing excitatory and inhibitory modulatory circuits, and (2) propranolol's effect on the two modulatory circuits is dose dependent: the high dose increasing spontaneous CeA activity by preferentially blocking an inhibitory circuit, the low dose decreasing spontaneous CeA activity by preferentially blocking an excitatory circuit, and the intermediate dose weakly suppressing CeA activity by blocking both the excitatory and inhibitory modulatory circuits. Disinhibition of CeA activity by the high dose of propranolol may explain the enhancement of retention observed in the passive-avoidance task when this dose of the drug is administered systemically, and may have implications for the use of propranolol clinically in treating aversive-memory-related anxiety disorders such as posttraumatic stress syndrome. PMID- 11420073 TI - Agmatine produces antinociception in tonic pain in mice. AB - Agmatine is an endogenous polyamine metabolite formed by decarboxylation of L arginine. In this study, the effect of agmatine on tonic pain was compared to its effect on phasic pain by using the formalin and tail-flick (TF) tests in mice. When administered intraperitoneally (ip), agmatine (37.5-300 mg/kg) exhibited a decrease in nociceptive behaviours in the first and second phase of the formalin test, which is a tonic pain model. The alpha(2) adrenoceptor antagonist yohimbine blocked the effect of agmatine in Phase 2 but did not change its effect in Phase 1. In the TF test, there was no significant change in the behaviour of agmatine administered (75-300 mg/kg) animals. As a result, agmatine appears to have an analgesic effect on tonic rather than phasic pain, and alpha(2) receptors seem partly to have a role in the antinociceptive effect of agmatine on tonic pain. PMID- 11420074 TI - Differential sensitivity to the anxiolytic effects of ethanol and flunitrazepam in PKCgamma null mutant mice. AB - Tests of ethanol effects in PKCgamma null mutant mice have indicated that PKCgamma plays a role in initial sensitivity to ethanol-induced sedation, hypothermia, and GABA(A) receptor function and impacts neurochemical pathways mediating anxiety. The present study was undertaken to evaluate whether the decreased sensitivity to ethanol previously observed in these mice generalized to the anxiolytic effects of ethanol. PKCgamma null mutant mice and wild-type controls were tested in the elevated-plus maze, the black/white box, and the mirrored chamber after ethanol (0, 1.0, 1.25, 1.5 g/kg) or flunitrazepam (FNZ) (0, 0.015, 0.03, 0.06 mg/kg). Results indicated that although both genotypes exhibited anxiolytic responses to ethanol in the elevated plus-maze, null mutant mice were less sensitive than wild-type control mice; however, in the black/white box, PKCgamma null mutants were more sensitive than controls to the anxiolytic effects of FNZ. Neither ethanol nor FNZ produced anxiolytic responses in the mirrored chamber for either genotype. These results suggest that PKCgamma differentially mediates anxiolytic responses to ethanol and FNZ and that this relationship interacts with each drug's efficacy in reducing anxiety-related behaviors specific to each of the three mazes. PMID- 11420075 TI - Nigrostriatal dopaminergic denervation enhances dopamine D(4) receptor binding in rat caudate-putamen. AB - Radioligand binding to dopamine (DA) D(4) receptors was examined in adult rat forebrain 5 weeks after unilateral 6-hydroxydopamine (6-OHDA) lesioning of substantia nigra to remove ascending nigrostriatal dopaminergic projections. D(4) receptor binding was increased by up to 47% in denervated caudate-putamen (CPu) in rats that rotated away from the lesioned side with apomorphine challenge, with lesser changes in rats that failed to rotate with apomorphine. Functional significance of D(4) receptor upregulation induced by the lesions was investigated by examining behavioral effects of the highly selective D(4) agonist CP-226,269 and antagonist CP-293,019. Neither agent induced rotation at doses as high as 30 mg/kg ip. Pretreatment with the D(4) antagonist CP-293,019 did not affect rotation induced by either a D(1)-like (SKF-38393) or D(2)-like receptor (quinpirole) agonist. These findings provide the first evidence that D(4) receptors can be upregulated by nigrostriatal dopaminergic denervation. They also suggest that, unlike D(1) and D(2) receptors, D(4) receptors do not play a pivotal role in rotational behavior in rats with unilateral dopaminergic lesions. PMID- 11420076 TI - The suppressive effects of intraperitoneal cocaine are augmented when evaluated in nondeprived rats. AB - Rats suppress intake of a saccharin conditioned stimulus (CS) when paired with all drugs of abuse tested including morphine, cocaine, heroin, amphetamine, and ethanol. Although most of these drugs suppress intake when administered via a range of routes, the efficacy of cocaine is an exception. Specifically, cocaine induced suppression of saccharin intake is much greater when administered subcutaneously than when administered intraperitoneally. The subcutaneous route of administration of cocaine, however, is somewhat problematic because, unless diluted, can cause stark necrosis. The present study, then, reexamined the effectiveness of intraperitoneal cocaine using less restrictive deprivation regimens that are known to facilitate the expression of the phenomenon. The results showed that, while only a 10- and 20-mg/kg dose of cocaine suppressed intake of the saccharin CS when evaluated in moderately water-deprived rats, all doses tested (i.e., 5, 10, and 20 mg/kg) significantly reduced CS intake when saccharin-cocaine pairings were evaluated in rats maintained on food and water ad libitum. Taken together, these data show that rats will readily avoid intake of a saccharin cue when paired with the intraperitoneal administration of cocaine and that the magnitude of the effect is augmented when examined in a need-free state. PMID- 11420077 TI - The effects of D-amphetamine on responding for candy and fruit drink using a fixed ratio and a progressive ratio schedule of reinforcer delivery. AB - The first purpose of this study was to compare the effects of D-amphetamine (AMPH) on operant responding reinforced under fixed ratio (FR) or progressive ratio (PR) schedules of reinforcement, testing the hypothesis that responding reinforced under a PR operant schedule would be disrupted by lower doses of AMPH than responding reinforced under a FR operant schedule. The second purpose of this study was to test the generalizability of the first hypothesis by comparing the effects of AMPH on responding reinforced by two different reinforcers under both FR and PR operant schedules. Rhesus monkeys had five to six candy and five to six fruit drink sessions per day, and could receive two reinforcers per session. Responding was initially reinforced under a PR procedure, such that the ratio size increased with each subsequent session. The parameters of the PR schedule were individually selected so that monkeys consumed a similar number of candy and fruit-drink reinforcers each day. The effects of oral AMPH (0.5, 0.75, 1.0 mg/kg) on responding were assessed. Responding was then stabilized using a FR schedule with parameters individually selected so that monkeys consumed a similar number of candy and fruit-drink reinforcers each day, and the effects of oral AMPH were again assessed. The PR breakpoint was significantly greater for candy than fruit-drink. AMPH produced dose-related decreases in both candy and fruit drink intake, but each AMPH dose decreased the number of fruit-drink deliveries to a greater extent than the number of candy deliveries. The results failed to support the hypothesis that responding under PR schedules of reinforcement would be disrupted by lower doses of AMPH. PMID- 11420078 TI - Extracellular dopamine in the rat prefrontal cortex during reward-, punishment- and novelty-associated behaviour. Effects of diazepam. AB - Variations of extracellular dopamine (DA(ext)) levels in prefrontal cortex were assessed by in vivo microdialysis. In rats trained in an operant fixed interval (FI(30s)) schedule of food delivery, acute exposure to contingent foot shocks resulted in a suppression of responding that was reversed by diazepam (4 mg/kg, ip). No changes in cortical DA(ext) levels occurred during this period in both control and treated rats. By contrast, in control rats, cortical DA(ext) levels increased (+25-40%) during the nonpunished component of the operant session, and during noncontingent food delivery (+25%). Control rats placed into an unfamiliar brightly lit openfield exhibited a marked increase in cortical DA(ext) levels (+100%). This effect occurred neither in rats given diazepam at a dose (2 mg/kg) which stimulated motor activity, nor during a second exposure to the openfield. In conclusion, a benzodiazepine-sensitive activation of mesoprefrontal DA neurones is induced by exposure to novel stressful surroundings and by food availability and consumption. The fact that cortical DA(ext) levels remained unchanged in rats that exerted complete control upon negative stimuli indicates that an activation of the mesoprefrontal DA system is not required for punishment induced behavioural blockade. PMID- 11420079 TI - Influence of social isolation and 6-OHDA lesion on the effects of quinelorane. AB - The sensitivity of the response to the preferential dopaminergic D3 (DAD3) receptor agonist, quinelorane, was compared in mice housed socially and in mice isolated for 4 weeks. Quinelorane (1, 5, 10, 50 and 100 microg/kg) was administered intraperitoneally. Motor activity was measured for 60 min posttreatment. Rectal temperature was measured prior to and 1 h following the administration of quinelorane (10, 50 and 100 microg/kg i.p.). Quinelorane significantly and dose-dependently decreased locomotor activity in social and in isolated mice. The locomotor activity of isolated mice was significantly lower than that of social mice, but isolation had no effect on quinelorane-induced hypomotility. Quinelorane decreased dose-dependently rectal temperature in isolated and social mice, but isolation had no effect on quinelorane-induced decrease in rectal temperature. The lesions of dopaminergic terminals with intracerebroventricular administration of 6-OHDA decreased the dopamine (DA) level by 93% in the nucleus accumbens and by 91% in the corpus striatum; these lesions impaired neither the hypolocomotion nor the hypothermia induced by quinelorane. Thus, it may be concluded that social isolation has no influence on the quinelorane-induced decreases in rectal temperature and in locomotor activity and that the DA receptors involved in these effects of quinelorane are located postsynaptically. PMID- 11420080 TI - Low-level lead exposure modulates effects of quinpirole and eticlopride on response rates in a fixed-interval schedule. AB - Exposure to low levels of lead (Pb) results in a wide range of behavioral changes. These behavioral deficits of lead are modified by duration of exposure, level of exposure, and stage of exposure. The mesoaccumbens dopamine (DA) system appears to be critically involved in these alterations; however, the precise mechanisms are not completely understood. This study investigated the effects of systemic administrations of the dopamine D(2)-like receptor agonist, quinpirole, and antagonist, eticlopride, on response rates of postweaning lead-exposed rats in a fixed-interval 1-minute (FI-1) schedule. Postweaning exposure to 50 ppm lead (lead acetate) resulted in increased response rates. The dopamine D(2)-like agonist, quinpirole (0.05, 1.0, 3.0 mg/kg), reversed the effects of lead by reducing the response rates. However the antagonist, eticlopride (0.01 and 0.05), did not produce any marked modulation of the response rates of the lead group. Rather, systemic injections of eticlopride attenuated the response rates of control rats. The effects suggest that lead-induced alterations in FI responding are modulated by dopamine D(2)-like mechanisms. Thus, postweaning, subchronic exposure to lead resulted in enhanced sensitivity to quinpirole administration and reduced sensitivity to eticlopride. These observations are consistent with attenuated dopaminergic activity. PMID- 11420081 TI - Lateral parabrachial lesions impair taste aversion learning induced by blood borne visceral stimuli. AB - The lateral parabrachial area (LPB), main relay from the area postrema (AP), plays a role in processing visceral information and is thus of potential importance in taste aversion learning (TAL). This study used a lesion approach to address whether LPB functional relevance depends upon the features of toxins that serves as visceral stimuli in TAL. In addition, we explored whether LPB involvement in TAL is restricted to those toxic events detected by the AP or whether it has a more general role. Results showed that LPB-lesioned animals were disrupted in acquiring a TAL induced by blood-borne AP-dependent aversive stimuli (intraperitoneal methylscopolamine) and by AP-independent stimulus (intraperitoneal ethanol), but still, clearly developed strong aversions when intragastric hypertonic sodium chloride, a vagally processed aversive stimulus, served as the aversive stimulus. These findings suggest that the LPB plays a critical role in TAL induced by blood-borne toxins, such as methylscopolamine or ethanol, but is not necessary for vagally mediated stimulus, such as sodium chloride. The present results are discussed in the context of the hypothesis holding separable and independent neural systems underlying TAL. PMID- 11420082 TI - Effects of metrifonate on the hippocampal theta rhythm of freely moving intact and MS-lesioned mice. AB - Changes in hippocampal electroencephalogram (EEG) have been suggested to be closely associated with spatial learning ability. Spatial learning can be improved in medial septal (MS)-lesioned mice by metrifonate, a cholinesterase inhibitor. We designed this study to investigate the effects of metrifonate on the hippocampal theta oscillation of intact and MS-lesioned mice. Intact and MS lesioned C57BL mice were treated with acute injections of metrifonate (doses: 15, 50 and 100 mg/kg ip). These included a dose that considerably improved spatial memory of MS-lesioned mice in our earlier study. In addition, subtype selective muscarinic agents, BIBN-99, AF267B and AF150(S) were used. Recordings of hippocampal theta during movement and awake immobility revealed a dramatic reduction of theta in the lesioned animals. Metrifonate induced prominent changes in the EEG of intact mice, but not of MS-lesioned mice. The effect of metrifonate was not mimicked by two selective M(1)-agonists and was augmented by a combined injection of a selective M(2)-antagonist. These data suggest that improved spatial learning by the cholinesterase inhibitor metrifonate is unrelated to its effects on the hippocampal EEG. These two effects may be mediated through different muscarinic receptor subtypes. PMID- 11420083 TI - Effects of neonatal handling on nociceptin/orphanin FQ and opioid peptide levels in female rats. AB - Animals exposed to short periods of handling during the critical period of development, i.e., the first 21 days of life in rats, show attenuated neuroendocrine responses to stress in adult life. We have previously reported long-term changes in brain dynorphin (DYN) peptide levels in male Sprague-Dawley rats after neonatal handling. The purpose of this study was to investigate whether neonatal handling, 15-min individual separation from the mother during postnatal days 1-21, can induce long-term changes in DYNB, Met-enkephalin Arg(6)Phe(7) (MEAP) and nociceptin/orphanin FQ (N/OFQ) immunoreactive (ir) levels in female Sprague-Dawley rats. The peptides were measured in brain and pituitary gland 2 months after the handling procedure. The results reveal that handled (H) rats had increased ir levels of N/OFQ, DYNB and MEAP in the periaqueductal gray (PAG) as compared to nonhandled (NH) controls. Furthermore, H rats had decreased ir levels of DYNB in the frontal cortex and in the amygdala. In contrast to previous findings in male rats, DYNB levels were unaffected in areas related to the hypothalamo-pituitary-adrenal (HPA)-axis. The results indicate that a manipulation early in life can induce persistent neurochemical changes in the N/OFQ and opioid peptide system in female Sprague-Dawley rats. PMID- 11420084 TI - Precipitated cannabinoid withdrawal is reversed by Delta(9)-tetrahydrocannabinol or clonidine. AB - The availability of the cannabinoid antagonist, SR 141716A, to precipitate withdrawal following repeated cannabinoid administration provides a model to investigate the mechanisms underlying cannabinoid dependence as well as potential treatments to alleviate withdrawal symptoms. The goal of the present study was to determine whether SR 141716A-precipitated withdrawal symptoms in Delta(9) tetrahydrocannabinol (Delta(9)-THC)-tolerant mice could be alleviated by either readministration of Delta(9)-THC or clonidine, an alpha(2)-receptor agonist. SR 141716A elicited paw tremors in Delta(9)-THC-tolerant mice, but produced a significant increase in head shakes independently of repeated Delta(9)-THC treatment. Readministration of Delta(9)-THC, following SR 141716A-precipitated withdrawal, reversed paw tremors (ED(50)=9.9 mg/kg), but failed to reduce head shaking behavior. Clonidine reversed SR 141716A-precipitated paw tremors (ED(50)=0.18 mg/kg) and blocked head shakes at all doses tested. The reversal effects did not appear to be the result of motor impairment because neither decreases in spontaneous locomotor activity nor motor incoordination, as assessed in the inverted screen test, could account for the effects. These findings suggest that SR 141716A precipitates paw tremors in mice by competing with Delta(9)-THC at the CB(1) receptor, though it also produced head shaking in nondependent animals. Finally, the observation that clonidine alleviated SR 141716A-precipitated paw tremors suggests its potential as a treatment for cannabinoid dependence. PMID- 11420085 TI - Potentiation of kappa-opioid receptor agonist-induced analgesia and hypothermia by fluoxetine. AB - The effect of fluoxetine, a selective 5-HT reuptake inhibitor on the analgesic and hypothermic response of trans-(+/-)-3,4-dichloro-N-methyl-N-[2-(1 pyrrolidinyl)-cyclohexyl]-benzeneacetamide methane sulphonate (U-50,488H) and (+/ )-trans-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl] benzo[b] thiophene-4-acetamide (PD 117302), kappa-opioid receptor agonists, was determined in female Sprague Dawley rats using the tail-flick method and telethermometer, respectively. Intraperitoneal injections of U-50,488H (U50) and PD 117302 (PD117) produced a dose-dependent analgesic and hypothermic response. Fluoxetine (10 mg/kg, i.p.) by itself did not produce an analgesic response. The analgesic response to U50 (10, 20, and 40 mg/kg, i.p.) and PD117 (7.5, 15, and 22.5 mg/kg, i.p.) was potentiated by fluoxetine injected intraperitoneally 60 min prior to the injection of kappa opioid agonists. Similarly, the hypothermic response of U50 (20 and 40 mg/kg, i.p.) and PD117 (7.5, 15, and 22.5 mg/kg, i.p.) was potentiated by fluoxetine. The results indicate that selective kappa-opioid receptor agonists-induced analgesia and hypothermia is potentiated by fluoxetine, suggesting the role of extracellular 5-HT in the kappa-opioid receptor-mediated analgesia and hypothermia. PMID- 11420086 TI - Biphasic effects of 7-OH-DPAT on the acquisition of responding for conditioned reward in rats. AB - Previous studies have shown that dopamine (DA) receptor subtype-specific agonists differentially affect responding for conditioned reward D1-like agonists impair, whereas D2-like agonists enhance responding. The present study compared the effects of the D2-like agonists bromocriptine and 7-hydroxy-N,N-di-n-propyl-2 aminotetralin (7-OH-DPAT). Food-deprived rats (N=159) were preexposed to a chamber with two levers, one producing a tone (3 s) and the other turning the house lights off (3 s), for five 40-min sessions. In four subsequent 65-min conditioning sessions with the levers removed, the lights-off stimulus was paired with food (80 presentations per session). During two 40-min test sessions, the lights-off (CR) and tone (NCR) levers were replaced and responses at each lever were recorded. Confirming previous results, bromocriptine (0.50-5.0 mg/kg) dose dependently enhanced responding on the lever producing conditioned reward. In contrast, 7-OH-DPAT had a biphasic effect on responding for conditioned reward. Low doses (0.10-0.25 mg/kg) reduced CR lever responding, whereas a higher dose of 1.0 mg/kg enhanced such responding. An intermediate dose of 0.50 mg/kg neither impaired nor enhanced CR lever responding. The biphasic profile of 7-OH-DPAT may arise through differential actions at D3 vs. D2 receptors or presynaptic vs. postsynaptic DA receptors at low and high doses, respectively. PMID- 11420087 TI - Differential actions of dopamine receptor antagonism in rats upon food intake elicited by either mercaptoacetate or exposure to a palatable high-fat diet. AB - Selective dopamine receptor antagonists have been shown to reduce food intake of rats under such regulatory challenge conditions as food deprivation and 2-deoxy-D glucose-induced glucoprivation, and under such palatable conditions as acute exposure to sucrose solutions. Food intake is increased following either pretreatment with the free fatty acid oxidation inhibitor, mercaptoacetate (MA), or acute exposure to a palatable high-fat source. The present study examined whether equimolar doses (50-800 nmol/kg, s.c.) of either the selective D(1) receptor antagonist, SCH23390, or the selective D(2) receptor antagonist, raclopride, would alter food intake elicited by either MA (70 mg/kg, i.p.) or acute exposure to a high-fat diet (67% ground rat chow, 33% vegetable shortening). SCH23390 significantly and dose-dependently reduced MA-induced feeding with the two higher (400 and 800 nmol/kg) doses eliminating this response after the first 2 h and the two lower (50 and 200 nmol/kg) doses preventing the occurrence of significant MA-induced feeding. Raclopride eliminated MA-induced feeding at the highest dose, and produced dose-dependent reductions at lower doses. A different pattern of dopamine antagonist effects emerged for high-fat intake. The identical dose range of SCH23390 failed to alter high-fat intake. In contrast, whereas the highest (800 nmol/kg) dose of raclopride significantly reduced high-fat intake after 1 h, the middle (200 and 400 nmol/kg) doses of raclopride significantly increased high-fat intake after 2 h. These data are discussed in terms of the modulatory actions of dopamine upon food intake, of the differential actions of dopamine receptor subtypes upon intake under challenge and palatable conditions, and of the potential participation of presynaptic and postsynaptic receptor populations in these responses. PMID- 11420088 TI - Nicotine alters some of cocaine's subjective effects in the absence of physiological or pharmacokinetic changes. AB - Tobacco smoking and cocaine use often co-occurs and the frequency of smoking has been positively correlated with the likelihood of cocaine use. In addition, nicotine pretreatment has been shown to increase the rate of cocaine self administration in rats and to enhance cue-induced cocaine craving in humans. The present study was conducted to investigate whether nicotine pretreatment via a transdermal patch alters the behavioral, physiological, and pharmacokinetic effects of an acute dose of cocaine in nondependent human volunteers. Seven male tobacco smokers who used cocaine occasionally provided informed consent and participated in this placebo-controlled, four-visit study. Following pretreatment with a transdermal nicotine patch (placebo, 14 mg), subjects were challenged with an acute dose of intranasal cocaine (placebo, 0.9 mg/kg). Nicotine pretreatment attenuated cocaine-induced increases in reports of "high" and "stimulated" and increased the latency to detect cocaine effects and cocaine-induced euphoria. Nicotine did not alter cocaine's effects on heart rate, skin temperature, and blood pressure or plasma cocaine, benzoylecgonine (BE), or ecgonine methylester (EME) concentrations. Our findings indicate that nicotine pretreatment alters some of the positive subjective effects of cocaine in humans without affecting cocaine's effects on physiologic responses or pharmacokinetic profiles. PMID- 11420089 TI - Differential behavioral responses to chronic amphetamine in adult male and female rats exposed to postnatal cocaine treatment. AB - The impact of cocaine exposure during development on behavioral sensitization as measured by locomotor activity and stereotypy following repeated intermittent administration of amphetamine is examined. Male and female Sprague-Dawley rats were exposed to cocaine at 50 mg/kg/day during postnatal days (PND) 11-20 and, as adults (PND193-212), were administered seven daily injections of 2.0 mg/kg amphetamine. Both locomotor activity and stereotypic behavior were assessed following the first and seventh injections. Control males and females showed sensitized behavior following repeated amphetamine injections with females showing greater locomotion while males showed increased stereotypy. Male rats pretreated with cocaine failed to develop sensitized locomotor or stereotypic responses following repeated amphetamine injections consistent with dampened D(1) receptor activity. Females pretreated with cocaine did not show a sensitized locomotor response but did display sensitization of stereotypy following repeated amphetamine administration. Thus, it appears that postnatal cocaine treatment produces differential effects on the circuits mediating sensitization behavior in male and female rats. PMID- 11420090 TI - Attenuation of cocaine-induced genomic and functional responses in prenatal cocaine-exposed rabbits. AB - The effects of in utero cocaine exposure on cocaine-induced genomic and functional responses in postnatal life were examined. Pregnant Dutch Belted rabbits were injected intravenously, twice daily, with cocaine hydrochloride (4 mg/kg) or saline from day 8 through day 29 of pregnancy. Prenatally exposed kits were challenged with cocaine on postnatal day 20. In prenatal saline-exposed kits, cocaine induced time- and dose-dependent c-fos gene expression in both frontal cortex and striatum. Prenatal cocaine exposure reduced cocaine-induced c fos responses by 35-58% in the frontal cortex and 37-41% in the striatum. Cocaine induced functional responses that included head bobbing, seizure, and locomotor activity were also attenuated in prenatal cocaine-exposed kits. Cocaine-induced c fos expression and functional responses were blocked by the D(1) dopamine receptor antagonist, SCH23390, or by the serotonin receptor antagonist, methysergide, but not by the D(2) dopamine receptor antagonist, L-sulpride. The results indicate that in utero cocaine exposure leads to diminished responses to cocaine challenge in the offspring, which may be mediated by prenatal cocaine induced alterations in one or more components of the D(1) dopamine and/or serotonin receptor signaling systems during early postnatal life. PMID- 11420091 TI - mu-Opioid receptor downregulation contributes to opioid tolerance in vivo. AB - The present study examined the contribution of downregulation of mu-opioid receptors to opioid tolerance in an intact animal model. Mice were implanted subcutaneously with osmotic minipumps that infused etorphine (50-250 microg/kg/day) for 7 days. Other mice were implanted subcutaneously with a morphine pellet (25 mg) or a morphine pellet plus an osmotic minipump that infused morphine (5-40 mg/kg/day) for 7 days. Controls were implanted with an inert placebo pellet. At the end of treatment, pumps and pellets were removed, and saturation binding studies were conducted in whole brain ([3H]DAMGO) or morphine and etorphine analgesic ED(50)s were determined (tail-flick). Morphine tolerance increased linearly with the infusion dose of morphine (ED(50) shift at highest infusion dose, 4.76). No significant downregulation of mu-receptors in whole brain was observed at the highest morphine treatment dose. Etorphine produced dose-dependent downregulation of mu-opioid receptor density and tolerance (ED(50) shift at highest infusion dose, 6.97). Downregulation of mu receptors only occurred at the higher etorphine infusion doses (> or =150 microg/kg/day). Unlike morphine tolerance, the magnitude of etorphine tolerance was a nonlinear function of the dose and increased markedly at infusion doses that produced downregulation. These results suggest that mu-opioid receptor downregulation contributes to opioid tolerance in vivo. Therefore, opioid tolerance appears to rely upon both "receptor density-dependent" and " receptor density-independent" mechanisms. PMID- 11420092 TI - Delta-9-tetrahydrocannabinol differentially suppresses cisplatin-induced emesis and indices of motor function via cannabinoid CB(1) receptors in the least shrew. AB - We have recently shown that the cannabinoid CB(1) receptor antagonist, SR 141716A, produces emesis in the least shrew (Cryptotis parva) in a dose- and route-dependent manner. This effect was blocked by delta-9-tetrahydrocannabinol (Delta(9)-THC). The present study investigates the cannabinoid receptor mechanisms by which Delta(9)-THC produces its antiemetic effects against cisplatin (20 mg/kg, i.p.)-induced emesis as well as its cannabimimetic activity profile (motor reduction) in the least shrew. Intraperitoneal administration of Delta(9)-THC (1, 2.5, 5 and 10 mg/kg) dose-dependently reduced both the percentage of animals vomiting (ID(50)=1.8+/-1.6 mg/kg) and the frequency of vomits (ID(50)=0.36+/-1.18 mg/kg) in a potent manner. The lowest significantly effective antiemetic dose of Delta(9)-THC for the latter emesis parameters was 2.5 mg/kg. Although Delta(9)-THC reduced the frequency of vomits up to 98%, it failed to completely protect all tested shrews from vomiting (80% protection). The cannabinoid CB(1) antagonist (SR 141716A) and not the CB(2) antagonist (SR 144528), reversed the antiemetic effects of Delta(9)-THC in a dose-dependent fashion. Delta(9)-THC (1, 5, 10 and 20 mg/kg, ip) suppressed locomotor parameters (spontaneous locomotor activity, duration of movement and rearing frequency) in a biphasic manner and only the 20-mg/kg dose simultaneously suppressed the triad of locomotor parameters to a significant degree. Subcutaneous (1-10 mg/kg) and intraperitoneal (0.05-40 mg/kg) injection of some doses of SR 141716A caused significant reductions in one or more components of the triad of locomotor parameters but these reductions were not dose dependent. Subcutaneous injection of SR 141716A (0.2, 1, 5 and 10 mg/kg) reversed the motor suppressant effects of a 20-mg/kg dose of Delta(9)-THC (ip) in a dose-dependent manner. Relative to its motor suppressant effects, Delta(9)-THC is a more potent antiemetic agent. Both effects are probably mediated via CB(1) receptors in distinct loci. PMID- 11420093 TI - Cannabinoid-induced motor incoordination through the cerebellar CB(1) receptor in mice. AB - Cannabinoids are known to impair motor function in humans and laboratory animals. We have observed dose-dependent motor incoordination in mice evaluated by rotorod following direct intracerebellar (i.c.b.) microinjection of synthetic cannabinoid agonists CP55,940 (5-25 microg) and HU-210 (1.56-6.25 microg), through permanently implanted stainless steel guide cannulas. The motor incoordination was marked at 15, 35 and 55 min post-microinjection. The motor incoordination elicited by HU-210 (6.25 microg) and CP55,940 (20 microg) was significantly blocked by the CB(1) receptor-selective antagonist SR141716A (25 microg i.c.b.), indicating mediation by a cerebellar CB(1) receptor. Further direct evidence of CB(1) mediation was obtained through a CB(1) receptor antisense/mismatch oligodeoxynucleotide approach (3 microg/12 h; total of six doses). Mice treated with intracerebellar antisense had a significantly diminished motor incoordination response to intracerebellar CP55,940 15 microg compared to mice that received intracerebellar mismatch or no prior treatment. Also, the response to intracerebellar CP55,940 in the CB(1) mismatch-treated mice did not differ from the mice that received only CP55,940. A separate study using a cerebellar tissue punching technique, following intracerebellar [3H]-CP55,940 microinjection, confirmed that cannabinoid drug dispersion following microinjections was exclusively confined to the cerebellum. Microinjection of CP55,940 (20 microg) into the hippocampus, an area with a large density of CB(1) receptors, did not impair motor coordination. Taken together, these results indicate that cannabinoid-induced motor impairment occurs by activation of a CB(1) receptor in the cerebellum. The participation of other brain motor areas in cannabinoid-induced motor incoordination will require future study. PMID- 11420094 TI - PMMA-stimulus generalization to the optical isomers of MBDB and 3,4-DMA. AB - Psychoactive phenylisopropylamines can produce one or more of several different stimulus effects in animals. These effects are typified by the hallucinogen 1 (2,5-dimethoxy-4-methylphenyl)-2-aminopropane (DOM), the central stimulant amphetamine, and by N-methyl-1-(4-methoxyphenyl)-2-aminopropane (PMMA), an agent whose actions are not yet well understood. The optical isomers of two phenylisopropylamines known to lack DOM and amphetamine-stimulus character, that is N-methyl-1-(3,4-methylenedioxyphenyl)-2-aminobutane (MBDB) and 1-(3,4 dimethoxyphenyl)-2-aminopropane (3,4-DMA), were examined in rats trained to discriminate 1.25 mg/kg of PMMA from vehicle. The PMMA stimulus (ED(50)=0.4 mg/kg) generalized to all four agents: S(+)-MBDB (ED(50)=0.8 mg/kg), R(-)-MBDB (ED(50)=2.0 mg/kg), S(+)-3,4-DMA (ED(50)=2.6 mg/kg) and R(-)-3,4-DMA (ED(50)=3.9 mg/kg). The results show that these agents produce stimulus effects similar to those produced by PMMA. Both isomers of MBDB have been previously demonstrated to substitute for N-methyl-1-(3,4-methylenedioxyphenyl)-2-aminopropane (MDMA) in rats trained to discriminate MDMA from vehicle, but MBDB-trained animals failed to recognize DOM or amphetamine. Similar results were obtained with the 3,4-DMA optical isomers in the present investigation using rats trained to discriminate MDMA, DOM or (+)-amphetamine from vehicle; both isomers of 3,4-DMA substituted for an MDMA stimulus, but not for a DOM or amphetamine stimulus. Taken together, the evidence suggests that PMMA, S(+)-MBDB, R(-)-MBDB, S(+)-3,4-DMA, R(-)-3,4 DMA, and S(+)-MDMA can produce common stimulus effects in rats. The present findings also better define the PMMA stimulus and the structural requirements necessary to produce this type of stimulus effect. PMID- 11420095 TI - Effects of sertraline on regional neuropeptide concentrations in olfactory bulbectomized rats. AB - Corticotropin-releasing factor (CRF) and thyrotropin-releasing hormone (TRH) are two neuropeptides that exhibit increased cerebrospinal fluid (CSF) concentrations during major depressive episodes while somatostatin (somatotropin-release inhibiting factor, SRIF) is decreased. Clinical and basic research findings indicate that clinically effective antidepressant therapies often normalize the indicators of CRF and TRH hypersecretion as well as SRIF hyposecretion. The olfactory bulbectomized (OBX) rat is used to screen potential antidepressant drugs for clinical efficacy. This model requires chronic administration of the antidepressant drug to normalize OBX-induced behaviors such as increased locomotion in a novel environment. This report describes the regional brain concentration changes in CRF, TRH and SRIF produced by OBX and demonstrates the ability of the selective serotonin re-uptake inhibitor and antidepressant drug, sertraline (10 mg/kg), to normalize certain of these alterations in regional neuropeptide concentrations as well as normalizing OBX-induced increases in locomotor activity. OBX-induced increases in CRF concentrations in the hypothalamus and bed nucleus of the stria terminalis were specifically and significantly decreased by sertraline. OBX-induced increases in TRH concentrations in the hypothalamus were reversed by sertraline. The concentration of SRIF was significantly reduced by OBX in the anterior caudate and the piriform cortex, but sertraline reversed these changes only in the anterior caudate. PMID- 11420096 TI - R-zacopride, a 5-HT3 antagonist/5-HT4 agonist, reduces sleep apneas in rats. AB - The effects of R-zacopride, a benzamide with potent 5-HT3 receptor antagonist and 5-HT4 receptor agonist properties, on spontaneous apneas were studied in 10 Sprague-Dawley rats by monitoring respiration and sleep for 6 h. R-zacopride (0.5, 1.0 and 10.0 mg/kg) suppressed spontaneous central apneas during non-rapid eye-movement (NREM) sleep by 50% (P=.05 for 0.5 mg/kg, P=.02 for 1.0 mg/kg and P=.001 for 10.0 mg/kg dose vs. control), and during rapid-eye-movement (REM) sleep by 80% by all doses tested (P<.0007) for at least 2 h after intraperitoneal injection. We conclude that R-zacopride, over a 20-fold dose range, significantly reduces central apnea expression during NREM and REM sleep in the rat. The efficacy of this compound to suppress central apneas most probably arises from its antagonist actions at 5-HT3 receptors or from its mixed agonist/antagonist profile at 5-HT4/5-HT3 receptors. PMID- 11420097 TI - Effects of insulin-produced hypoglycemia in combination with ethanol on spontaneous motor activity in rats. AB - The hazardous consequences of drinking alcohol by persons receiving insulin treatment is indicated by clinical reports, but little controlled research has investigated the combination of hypoglycemia and ethanol intoxication. Ethanol's effect on spontaneous motor activity (SMA, detected by Opto-Varimax activity meters) in hypoglycemic (HG) rats was determined over a range of ethanol doses in two experiments. Combinations of insulin and ethanol were administered intraperitoneally to moderately food-deprived rats. Blood glucose was measured before and after a 30-min SMA-monitoring period. In Experiment 1, ethanol doses of 300, 600 and 1200 mg/kg were combined with insulin at doses 0.5 and 2 U/kg. A second experiment tested a narrower range of drug doses (ethanol 600 and 1200 mg/kg, insulin 1 U/kg) under slightly different procedures. After insulin treatment, blood-glucose levels dropped to approximately 40-60% of control levels and this HG was accompanied by decreased SMA. Ethanol did not influence blood glucose levels, nor did it potentiate the HG produced by insulin. Combination of HG and the highest ethanol dose potentiated the SMA-depressant effect in both experiments, whereas lower ethanol doses partially reversed the suppression of motor activity in HG rats. PMID- 11420098 TI - Progesterone treatment during the early follicular phase of the menstrual cycle: effects on smoking behavior in women. AB - The goals of this study were (1) to examine the feasibility of administering progesterone to women during the early follicular phase when the endogenous estradiol and progesterone levels are low, and (2) to investigate the effects of oral progesterone treatment on smoking behavior in female smokers. Twelve subjects had two experimental sessions, within 3-9 days after the beginning of their menses. In each experimental session, subjects received a single 200-mg dose of progesterone or placebo, orally. Two and a half hours after the medication treatment, subjects were assessed for subjective response to two puffs of a cigarette and then started the self-administration period in which they had the option to exchange their token for two puffs of cigarette, 15 min apart. Subjects had low levels of estradiol and progesterone before the first and second sessions. Plasma progesterone levels peaked in 2 h following progesterone treatment. Progesterone treatment attenuated the craving for and subjective effects from smoking. Under progesterone treatment, there was a trend for decreased smoking behavior. These preliminary results suggest that the early follicular phase of the menstrual cycle may be a useful interval to investigate the effects of exogenous progesterone in female smokers. The effects of progesterone on nicotine dependence need to be studied further. PMID- 11420099 TI - Perceptual and motor effects of morphine and buprenorphine in baboons. AB - The effects of morphine and buprenorphine on auditory perceptual discriminations and response latency ("reaction time") in baboons are compared. The task employed synthetic human vowel sounds that are readily generated in the laboratory, and closely approximate natural baboon "grunt" vocalizations [J. Acoust. Soc. Am. 101 (1997) 2951]. Baboons pressed a lever to produce one repeating "standard" vowel, and released the lever only when one of four other "comparison" vowels occasionally occurred in place of the standard vowel. The percentage of correct detections and median reaction time for each comparison were measured following intramuscular drug administrations of morphine (0.01-1.8 mg/kg) and buprenorphine (0.00032-0.032 mg/kg). Both morphine and buprenorphine impaired vowel discriminability, and greater impairments occurred for those comparison vowels that were more similar in formant structure to the standard vowel. Morphine increased reaction time in all baboons, and buprenorphine increased reaction time in two of three baboons. Morphine's perceptual effects occurred within 20-40 min following drug administration; buprenorphine's perceptual effects occurred 50-100 min following drug administration. Morphine and buprenorphine did not differ in the time course of their maximal reaction time effects. The results demonstrate that both morphine and buprenorphine can impair auditory discriminations involving human vowel sounds in baboons, as well as lengthen reaction times to the stimuli. PMID- 11420100 TI - Molecules on the surface of the Plasmodium falciparum infected erythrocyte and their role in malaria pathogenesis and immune evasion. AB - The surface of the erythrocyte undergoes a number of modifications during infection by Plasmodium falciparum. These modifications are critical for pathogenesis of severe disease and the acquisition of host immunity through their role in interactions between the host and the parasite and in antigenic variation. Our knowledge of the molecular basis for these processes has increased dramatically over the last few years, through a combination of genomic and biochemical studies. This review provides a summary of the molecules involved in cytoadherence and antigenic variation in P. falciparum. PMID- 11420101 TI - Extreme geographical fixation of variation in the Plasmodium falciparum gamete surface protein gene Pfs48/45 compared with microsatellite loci. AB - Comparing patterns of genetic variation at multiple loci in the genome of a species can potentially identify loci which are under selection. The large number of polymorphic microsatellites in the malaria parasite Plasmodium falciparum are available markers to screen for selectively important loci. The Pfs48/45 gene on Chromosome 13 encodes an antigenic protein located on the surface of parasite gametes, which is a candidate for a transmission blocking vaccine. Here, genotypic data from 255 P. falciparum isolates are presented, which show that alleles and haplotypes of five single nucleotide polymorphisms (SNPs) in the Pfs48/45 gene are exceptionally skewed in frequency among different P. falciparum populations, compared with alleles at 11 microsatellite loci sampled widely from the parasite genome. Fixation indices measuring inter-population variance in allele frequencies (F(ST)) were in the order of four to seven times higher for Pfs48/45 than for the microsatellites, whether considered (i) among populations within Africa, or (ii) among different continents. Differing mutational processes at microsatellite and SNP loci could generally affect the population structure at these different types of loci, to an unknown extent which deserves further investigation. The highly contrasting population structure may also suggest divergent selection on the amino acid sequence of Pfs48/45 in different populations, which plausibly indicates a role for the protein in determining gamete recognition and compatibility. PMID- 11420102 TI - Fatty acid remodeling of glycosyl phosphatidylinositol anchors in Trypanosoma brucei: incorporation of fatty acids other than myristate. AB - Trypanosoma brucei is the protozoan parasite that causes African sleeping sickness. Its surface is packed with 10(7) copies of the glycosyl phosphatidylinositol (GPI)-anchored variant surface glycoprotein (VSG). This GPI anchor is unusual in that it contains two myristates (14:0) in its lipid moiety. This fatty acid specificity is achieved through myristoylation of the GPI precursor, and the acyltransferases involved in the GPI remodeling were presumed to be specific for myristate. However, their specificity had never been fully evaluated. Here we found as expected that the remodeling acyltransferases completely excluded palmitate (16:0) and stearate (18:0) in a cell-free fatty acid remodeling system. In contrast, we found surprisingly that one of these enzymes was permissive to shorter fatty acids such as laurate (12:0) and octanoate (8:0). However, the rates of incorporation of shorter fatty acids were lower than that of myristate at low substrate concentration. Since shorter fatty acids are virtually absent in the parasite and in the host bloodstream, it is unlikely that shorter fatty acids compete effectively with myristate as remodeling substrates under physiological conditions. Even if they were present in small quantities, a recently identified specialized fatty acid synthetase efficiently elongates shorter fatty acids to myristate prior to incorporation into GPIs (Morita et al., Science 288 (2000) 140-3.). Therefore, even though a remodeling acyltransferase is permissive with regard to substrate chain length, the myristate specificity in GPI anchors is very high. PMID- 11420103 TI - Defining the cell cycle for the tachyzoite stage of Toxoplasma gondii. AB - Tachyzoite endodyogeny is characterized by a three phase cell cycle comprised of major G1 and S phases with mitosis following immediately upon the conclusion of DNA replication. Cytokinesis, which begins with the formation of daughter apical complexes, initiates in late S phase and overlaps mitosis. There is no evidence to support an extended G2 period in these parasites. In all strains, parasites with a 2 N DNA content are a relatively small subpopulation and when tachyzoites expressing a fluorescent nuclear marker (green-fluorescent-protein fused to proliferating-cell-nuclear-antigen) were observed by time-lapse microscopy, there appeared to be little delay between S phase and mitosis. Measurements of the DNA content of RH parasites by flow cytometry demonstrated that the G1 and S periods were approximately 60 and approximately 30% of a single division cycle, although these phases were longer in strains that display a slower growth rate. The overall length of S phase was determined by [3H]-thymidine autoradiography using transgenic parasites expressing herpes simplex thymidine kinase and validated by Northern analysis of S phase specific genes during synchronous growth. The fraction of S phase parasites by flow cytometry paralleled autoradiography, however, within S phase, the distribution of parasites was bimodal in all strains examined. Parasites containing a 1-1.7 N DNA complement were a small fraction when compared to the major S phase population which contained a near-diploid ( approximately 1.8 N) complement, suggesting parasites in late S phase have a slower rate of DNA replication. In lieu of a short or missing G2, where checkpoints are thought to operate in other eukaryotes, the bimodal replication of tachyzoite chromosomes may represent a distinct premitotic checkpoint associated with endodyogeny. PMID- 11420104 TI - The Trypanosoma brucei mitochondrial ATP synthase is developmentally regulated at the level of transcript stability. AB - The mitochondrial ATP synthase is developmentally regulated throughout the life cycle of the Trypanosoma brucei. The alpha and beta subunits of the F(1) moiety, and subunit 9 of the F(0) moiety of the T. brucei ATP synthase have been previously cloned and characterized. Here we have determined the chromosomal localization and developmental regulation of these three key subunits of the complex. Southern blot analysis indicates that all three of these genes are present as single copies in the T. brucei genome. Pulsed field gel analysis demonstrates that these genes are encoded in different chromosomes, and are thus not part of the same gene cluster. A comparison between the protein and steady state transcript levels for these subunits suggests that regulation of expression occurs predominantly posttranscriptionally. Comparison of mRNA stability for procyclic and bloodstream forms shows that the half life of the three transcripts is much shorter in bloodstream forms. The differences in transcript stability in the procyclic form for subunit 9 is greater than that for alpha and beta subunits, while the differences at the protein levels are comparable. These results suggest that there may be further posttranscriptional regulation of subunit 9. PMID- 11420105 TI - Ovothiol and trypanothione as antioxidants in trypanosomatids. AB - The relative amounts of ovothiol A (N(1)-methyl-4-mercaptohistidine) and trypanothione [N(1),N(8)-bis(glutathionyl)spermidine] have been determined in all life cycle stages of representative trypanosomatids (Leishmania spp, Crithidia fasciculata, Trypanosoma cruzi and T. brucei). Ovothiol A is present in all insect stages with intracellular concentrations of >1 mM for five species of Leishmania promastigotes and <0.25 mM for other trypanosomatids. In Leishmania promastigotes, ovothiol A can exceed trypanothione content particularly in late logarithmic and stationary phases of growth. In the other trypanosomatids, it represents less than 10% of the total thiol pool. Although amastigotes of L. major and L. donovani contain equivalent amounts of glutathione and trypanothione, ovothiol A is present in the former but absent in the latter. Ovothiol A is present in all developmental stages of T. cruzi but absent in bloodstream trypomastigotes of T. brucei. No ovothiol reductase activity could be detected in dialysed parasite extracts. Ovothiol disulphide is not a substrate for trypanothione reductase, although it can be reduced by the concerted action of trypanothione and trypanothione reductase. No ovothiol-dependent peroxidase activity was present in Leishmania extracts. Although ovothiol A can act as a non enzymatic scavenger of hydrogen peroxide, it is less efficient than trypanothione. Second order rate constants were determined with trypanothione>glutathionylspermidine>ovothiol>glutathione. Given the presence of an active trypanothione peroxidase system in all these trypanosomatids, it is concluded that under physiological conditions, ovothiol is unlikely to play a major role in the metabolism of hydrogen peroxide in intact cells. Nonetheless, since ovothiol is absent in host macrophage, kidney and CHO cells, this metabolite may have other important functional roles in trypanosomatids that could be exploited as a chemotherapeutic target. PMID- 11420106 TI - Secretion of the novel Trichinella protein TSJ5 by T. spiralis and T. pseudospiralis muscle larvae. AB - The Trichinella tsJ5 gene is preferentially expressed in muscle larvae of Trichinella spiralis and encodes a novel protein. Previous observations have shown tsJ5 to be expressed at higher levels in encapsulating species than in non encapsulating species and down-regulation of gene expression in T. pseudospiralis to be correlated with a lower protein abundance in the muscle larva of this species. In the present study we have determined the full-length cDNA sequence of the tsJ5 homologue in T. pseudospiralis (tpJ5). Antigens recognised by an anti-J5 antibody are found on the cuticular surface of both T. spiralis and T. pseudospiralis muscle larvae, as well as in the body wall muscle. We show that both the TSJ5 and TPJ5 proteins are found in the excretory/secretory fractions collected from muscle larva cultured in vitro and that despite the absence of a typical N-terminal signal sequence, secretion of pTSJ5 is mediated through the classical ER/Golgi secretory pathway. PMID- 11420107 TI - Identification and characterisation of a RAD51 gene from Leishmania major. AB - The RAD51 gene is a homologue of Escherichia coli recA which plays a central role in homologous recombination and DNA repair. This paper describes the identification of the RAD51 gene from the trypanosomatid parasite Leishmania major. The LmRAD51 gene codes for a 377 amino acid polypeptide with a predicted molecular mass of 41259 Da that is highly homologous to the Rad51 family of proteins. Recombinant L. major Rad51 protein (LmRad51) was over-expressed in a bacterial expression system, purified to homogeneity and shown to bind DNA and exhibit DNA-stimulated ATPase activity, consistent with previously reported biochemical characteristics of Rad51 protein. Although LmRad51 expression is below the level of detection in exponentially growing cultures of Leishmania, high levels of LmRad51 mRNA and protein expression can be detected following exposure to the DNA-damaging agent phleomycin. LmRAD51 is one of the first examples of a DNA damage-inducible gene to be characterised in Leishmania, and will be invaluable in studying the contribution of homologous recombination to Leishmania virulence. PMID- 11420108 TI - Genetic analysis of spermidine synthase from Leishmania donovani. AB - The polyamine biosynthetic pathway of protozoan parasites has been validated as a target in antiparasitic chemotherapy. To investigate this pathway at the biochemical and genetic level in a model parasite, the gene encoding spermidine synthase (SPDSYN), a key polyamine biosynthetic enzyme, has been cloned and sequenced from Leishmania donovani. The L. donovani SPDSYN gene encodes a polypeptide of 300 amino acids that exhibits 56% amino acid identity with the human counterpart. SPDSYN is present as a single copy gene in the leishmanial genome and encodes a 1.6 kb transcript. Employing SPDSYN flanking sequences to construct drug resistance cassettes, a Deltaspdsyn knockout strain of L. donovani was created by double targeted gene replacement. This Deltaspdsyn line could not convert putrescine to spermidine and was auxotrophic for polyamines. The polyamine auxotrophy could be circumvented by exogenous spermidine but not by putrescine (1,4-diaminobutane), cadaverine (1,5-diaminopentane), 1,3 diaminopropane, or spermine. Incubation of the null mutant in polyamine-deficient medium resulted in a rapid depletion in the intracellular spermidine level with a concomitant elevation of the putrescine pool. In addition, the level of trypanothione, a spermidine-containing thiol, was reduced, whereas the glutathione pool increased 3-4-fold. These data establish that SPDSYN is an essential enzyme in L. donovani promastigotes. The molecular and cellular reagents created in this investigation provide a foundation for subsequent structure-function and inhibitor design studies on this key polyamine biosynthetic enzyme. PMID- 11420109 TI - The lysosomal targeting and intracellular metabolism of trypanosome lytic factor by Trypanosoma brucei brucei. AB - Trypanosome lytic factor (TLF) provides innate protection for humans against infection by the animal pathogen Trypanosoma brucei brucei but not against the agent of human African sleeping sickness, Trypanosoma brucei rhodesiense. TLF exists in two forms, TLF-1 and TLF-2. Prior studies suggested that TLF-1 causes lysosomal disruption and subsequent cell death in T. b. brucei. Here we confirm the lysosomal targeting of TLF-1 by immunolocalization with the trypanosome lysosomal membrane protein p67, and by co-fractionation of radiolabelled TLF-1 with lysosomal enzymes. In addition, pulse-chase studies indicate that TLF-1 is not degraded within the lysosome as compared to the host protein transferrin. In TLF-1 treated cells, transferrin is degraded normally, indicating that lysosomal proteases remain active during the early phase of TLF-1 treatment but fail to degrade TLF-1. Following endocytosis a TLF lipoprotein appears to undergo disulfide bond reduction prior to entering the lysosome. Results presented here indicate that TLF-1 lipoproteins are targeted to the lysosome but are resistant to trypanosome lysosomal proteases. PMID- 11420110 TI - Unusual diversity in alpha-amanitin sensitivity of RNA polymerases in trichomonads. AB - Previous studies in the parasitic protist Trichomonas vaginalis have revealed that protein coding genes are transcribed by an alpha-amanitin-resistant RNA polymerase (RNAP) II. To investigate whether this unusual property is a general characteristic of trichomonads, we addressed the physiology of RNA synthesis in lysolecithin-permeabilized cells. Unlike in T. vaginalis, RNAP II in Tritrichomonas foetus was highly sensitive to the inhibitor alpha-amanitin. On the other hand, RNAP III, identified by its sensitivity to the specific inhibitor tagetitoxin, was found to be resistant to alpha-amanitin in Tritrichomonas foetus, but showed a typical intermediate sensitivity in T. vaginalis. Extension of this study to an additional seven trichomonad species confirmed this genera specific pattern of alpha-amanitin sensitivity and highlighted an unusual diversity in RNAPs among trichomonads, a closely related group of unicellular eukaryotes. PMID- 11420111 TI - Characterization of poly(ADP-ribose)polymerase from Crithidia fasciculata: enzyme inhibition by beta-lapachone. AB - Crithidia fasciculata poly(ADP-ribose)polymerase (PARP) has been isolated and partially purified. This is the first PARP isolated from trypanosomatids; it requires DNA and histone for activity, using NAD(+) as substrate. Thiol compounds specially dithiothreitol essentially contributed to PARP stability during purification and to PARP activity during assays. Nicotinamide, 3-aminobenzamide, theophylline, histamine, histidine, N-ethylmaleimide, p-chloromercuribenzoic acid, p-chloromercuriphenylsulfonic acid and o-iodosobenzoate inhibited PARP, thus confirming enzyme identity. PARP was also inhibited by the Fe(II)/H(2)O(2) Fenton system. beta-Lapachone inhibited PARP, apparently by direct interaction with the enzyme. PMID- 11420112 TI - Characterization of the subpellicular network, a filamentous membrane skeletal component in the parasite Toxoplasma gondii. AB - Electron microscopic examination of detergent-extracted Toxoplasma tachyzoites reveals the presence of a mechanically stable cytoskeletal structure associated with the pellicle of this parasite. This structure, composed of interwoven 8-10 nm filaments, is associated with the cytoplasmic face of the pellicle and surrounds the microtubule-based cytoskeleton. Two protein components of this network, TgIMC1 and TgIMC2, were identified. Both are novel proteins, but have a resemblance to mammalian filament proteins in that they are predicted to have extended, coiled-coil domains. TgIMC1 is also homologous to articulins, the major components of the membrane skeleton of algae and free-living protists. A homologue of TgIMC1 in the related malaria parasite Plasmodium falciparum was also identified suggesting the presence of structurally similar membrane skeletons in all apicomplexan parasites. We suggest that the subpellicular network, formed by TgIMC1 and 2 in Toxoplasma gondii and related parasites, plays a role in the determination of cell shape and is a source of mechanical strength. PMID- 11420113 TI - Cloning, nucleotide sequencing and analysis of the gene encoding the glutamate rich protein (GLURP) from Plasmodium reichenowi. PMID- 11420114 TI - Uptake and hydrolysis of sphingomyelin analogues in Plasmodium falciparum infected red cells. PMID- 11420115 TI - Analysis of the 18S rRNA genes of Sarcocystis species suggests that the morphologically similar organisms from cattle and water buffalo should be considered the same species. PMID- 11420116 TI - Palmitoylation of the canine histamine H2 receptor occurs at Cys(305) and is important for cell surface targeting. AB - To determine the presence and functional role of the histamine H2 receptor (H2R) palmitoylation, a receptor with a Cys(305) to Ala (A(305) receptor) mutation was generated. Wild-type (WT) and A(305) receptors were tagged at their N-termini with a hemagglutinin (HA) epitope. WT, but not A(305), receptors incorporated [3H]palmitate by metabolic labeling, indicating that the H2R is palmitoylated at Cys(305). Immunocytochemistry of WT and A(305) receptors expressed in COS7 cells revealed WT receptors to be distributed at the plasma membrane, while the majority of A(305) receptors were localized intracellularly with only a small portion being at the plasma membrane. However, the affinity of the A(305) receptor for tiotidine was comparable to that of the WT receptor. In addition, when the amounts of cell surface receptors as determined by anti-HA antibody binding were equivalent, A(305) receptors mediated production of more cAMP than WT receptors. Preincubation of COS7 cells expressing each receptor with 10(-5) M histamine for 30 min reduced subsequent cAMP production in response to histamine via the receptors to similar extents, indicating that palmitoylation is not necessary for desensitization. In addition, cell surface A(305) receptors were capable of being internalized from the cell surface at a rate and extent similar to those of WT receptors. Finally, CHO cell lines stably expressing either WT or A(305) receptors were incubated with 10(-5) M histamine for 1, 6, 12 and 24 h. Total amounts of WT and A(305) receptors, as determined by tiotidine binding, were reduced by incubation, indicating downregulation. Downregulation of the A(305) receptor was more extensive than that of the WT receptor. Thus, palmitoylation of the H2R might be important for targeting to the cell surface and stability. PMID- 11420117 TI - NADH oxidase activity (NOX) and enlargement of HeLa cells oscillate with two different temperature-compensated period lengths of 22 and 24 minutes corresponding to different NOX forms. AB - NOX proteins are cell surface-associated and growth-related hydroquinone (NADH) oxidases with protein disulfide-thiol interchange activity. A defining characteristic of NOX proteins is that the two enzymatic activities alternate to generate a regular period length of about 24 min. HeLa cells exhibit at least two forms of NOX. One is tumor-associated (tNOX) and is inhibited by putative quinone site inhibitors (e.g., capsaicin or the antitumor sulfonylurea, LY181984). Another is constitutive (CNOX) and refractory to inhibition. The periodic alternation of activities and drug sensitivity of the NADH oxidase activity observed with intact HeLa cells was retained in isolated plasma membranes and with the solubilized and partially purified enzyme. At least two activities were present. One had a period length of 24 min and the other had a period length of 22 min. The lengths of both the 22 and the 24 min periods were temperature compensated (approximately the same when measured at 17, 27 or 37 degrees C) whereas the rate of NADH oxidation approximately doubled with each 10 degrees C rise in temperature. The rate of increase in cell area of HeLa cells when measured by video-enhanced light microscopy also exhibited a complex period of oscillations reflective of both 22 and 24 min period lengths. The findings demonstrate the presence of a novel oscillating NOX activity at the surface of cancer cells with a period length of 22 min in addition to the constitutive NOX of non-cancer cells and tissues with a period length of 24 min. PMID- 11420118 TI - Block copolymers modify the internalization of micelle-incorporated probes into neural cells. AB - An important therapeutic concern is rate and extent of internalization of drugs into cells. Hydrophilic agents often internalize poorly and slowly, and highly lipophilic ones too rapidly. The incorporation of drugs into micelles allows regulation of their internalization parameters, and newly-described block copolymers can be selectively tailored to suit specific drugs. This report compares internalization of Cell Tracker CM-DiI (DiI), a highly lipophilic non cytotoxic fluorescent probe in common use in biology, from the freely-presented (non-micelle-incorporated) and micelle-incorporated states. DiI was effectively incorporated (>60%) into 25-50 nm diameter spherical micelles made from polycaprolactone-b-polyethylene oxide block copolymer. Confocal microscopy was used to evaluate the internalization of DiI into mixed neuron-glia cultures (2-14 days in vitro, 2DIV-14DIV). Incorporation of DiI into micelles strikingly reduced the rate and extent of its internalization in both 2DIV and 14DIV cultures. Both the age of the cultures and the block copolymer employed to construct the micelles significantly influence the internalization of micelle-incorporated probe. PMID- 11420119 TI - Colchicine inhibits taurodeoxycholate transport in pericentral but not in periportal hepatocytes. AB - Indirect evidence for a microtubule-dependent vesicular hepatocellular transport of bile acids has accumulated. Since inhibition of this transport by colchicine can be achieved only at high but not at low bile acid infusion rates we were wondering whether this transport pathway shows a hepatic zonation or not. To answer this question we perfused isolated rat livers antegradely or retrogradely, respectively, with unlabeled and labeled taurocholate or taurodeoxycholate. Inhibition of microtubule-dependent bile acid transport was aimed at co-infusion of colchicine. Periportal cells eliminated the likewise hydrophobic taurodeoxycholate as fast as the more hydrophilic taurocholate. In contrast, pericentral cells excreted taurodeoxycholate much slower than taurocholate. Colchicine did not change the biliary taurocholate excretion profile in periportal and pericentral cells. However, colchicine reduced significantly taurodeoxycholate excretion in pericentral but not in periportal cells. It is concluded that a microtubule-dependent vesicular, colchicine-sensitive transport pathway seems to be involved in the translocation of taurodeoxycholate in pericentral but not in periportal cells. Since such a vesicular bile acid transport is regarded to be much slower than transcellular transport by diffusion, this observation may explain the much slower excretion of hydrophobic bile acids like taurodeoxycholate in pericentral than in periportal cells under physiological conditions. PMID- 11420120 TI - Suppression of transmitter release by Tat HPC-1/syntaxin 1A fusion protein. AB - It has been reported that the fusion protein with the protein transduction domain (PTD) peptide of HIV-1 Tat protein can be internalized through the cell membrane of intact cells, although the exact mechanism is unknown. In this report, we investigated whether this new method could be used for the molecular analysis of exocytosis via HPC-1/syntaxin 1A, which plays an important role in transmitter release. When applied to PC12 cells, Tat PTD fusion proteins were rapidly internalized into most cells. In order to show that the internalized protein remained biologically active, the H3 domain of HPC-1/syntaxin 1A was fused to Tat PTD (Tat-H3). Transmitter release in PC12 cells was suppressed by Tat-H3 treatment. These results indicate that the Tat fusion protein is a useful tool for analyzing the process of transmitter release. PMID- 11420121 TI - Inhibition of the Na(+)/H(+) antiporter suppresses IL-12 p40 production by mouse macrophages. AB - The amiloride-inhibitable Na(+)/H(+) antiporter plays an important role in macrophage activation. The intracellular pathways leading to interleukin (IL)-12 p40 production by activated macrophages are incompletely understood. In the present study, we examined the contribution of the Na(+)/H(+) antiporter to the production of IL-12 p40. Amiloride or its analogs decreased the production of IL 12 p40 in macrophages stimulated with bacterial lipopolysaccharide and interferon gamma. The order of potency of amiloride analogs was consistent with the proposition that the effect of amiloride is mediated by the inhibition of the Na(+)/H(+) antiporter. The effect of amiloride was post-transcriptional, as IL-12 p40 mRNA levels induced by lipopolysaccharide and interferon-gamma were not affected by this inhibitor. Furthermore, the inhibitory effect of amiloride on IL 12 p40 production was not a result of interference with the activation of the p38 and p42/44 mitogen-activated protein kinases or c-Jun kinase. In summary, the production of IL-12 p40 requires a functional Na(+)/H(+) antiporter. PMID- 11420122 TI - Expression of transient receptor potential mRNA isoforms and Ca(2+) influx in differentiating human stem cells and platelets. AB - Store-regulated Ca(2+) entry (SOCE) is an important mechanism of elevating cytosolic [Ca(2+)]i in platelets, though the Ca(2+) influx channels involved are still unclear. We screened human platelets and their precursor cells (human stem cells and megakaryocytes) for the presence of candidate influx channels, i.e., isoforms of the Trp family of proteins. Primary stem cells were cultured with thrombopoietin to allow differentiation into megakaryocytes. The undifferentiated stem cells (CD34(+)) showed mRNA expression of only a spliced variant Trp1A. Immature (CD61(+)/CD42b(low)) and mature (CD61(+)/CD42b(high)) megakaryocytes as well as platelets expressed in addition unspliced Trp1 as well as Trp4 (less abundant) and Trp6 isoforms. This unspliced isoform appeared to be specific for cells of the megakaryocyte/platelet lineage, since immature (CD14(+)/CD61( )/CD42b(-)) and mature monocytes expressed only the Trp1A isoform. This conclusion was confirmed by the presence of Trp1A, 3, 4 and 6 transcripts in the immature megakaryocytic Dami cell line, and of Trp1, 1A, 4 and 6 transcripts in the more mature CHRF-288 cell line. The up-regulation of Trp1, 4 and 6 in the lineage from primary stem cells to mature megakaryocytes and platelets was accompanied by increased influx of extracellular Ca(2+) after pretreatment of the cells with thapsigargin or thrombin. Expression of new Trp isoforms in the differentiated cells is thus accompanied by increased SOCE. PMID- 11420123 TI - Inhibition of phosphatidylserine synthesis in Jurkat T cells by hydrogen peroxide. AB - Incubation of Jurkat cells in the presence of H2O2 either directly added to the culture medium or generated with glucose oxidase, menadione or the couple xanthine/xanthine oxidase induced a marked decrease of phosphatidylserine synthesis in the absence of changes in the synthesis of phosphatidylcholine and phosphatidylethanolamine. Concentration dependent response curves indicated that H2O2 induced inhibition of phosphatidylserine synthesis with an IC(50)=5 microM while both induction of tyrosine phosphorylation of proteins and Ca(2+) signals were obtained with an EC(50)=300 microM. The tyrosine kinase and Ca(2+) independent mechanism was confirmed by comparing the H2O2-induced and the CD3 induced inhibition of phosphatidylserine synthesis using several Jurkat clones differing in the expression of cell surface receptors such as CD3/TCR and CD45 and protein tyrosine kinase such as p72syk, ZAP-70 and p56lck. While CD3-induced inhibition of phosphatidylserine synthesis necessitates protein tyrosine phosphorylation and Ca(2+) signals, H2O2 provoked its effect in all the clones studied independently of the presence or absence of the proteins previously shown to be key elements in T cell signal transduction. Conversely, the antioxidant molecule, butylated hydroxanisole, generates an increased PtdSer synthesis, suggesting that the synthesis of this phospholipid is regulated by the redox status of the cells. PMID- 11420125 TI - Testis determination in mammals: more questions than answers. AB - In humans, testis development depends on a regulated genetic hierarchy initiated by the Y-linked SRY gene. Failure of testicular determination results in the condition termed 46,XY gonadal dysgenesis (GD). Several components of the testis determining pathway have recently been identified though it has been difficult to articulate a cascade with the known elements of the system. It seems, however, that early gonadal development is the result of a network of interactions instead of the outcome of a linear cascade. Accumulating evidence shows that testis formation in man is sensitive to gene dosage. Haploinsufficiency of SF1, WT1 and SOX9 is responsible for 46,XY gonadal dysgenesis. Besides, data on SRY is consistent with possible dosage anomalies in certain cases of male to female sex reversal. 46,XY GD due to monosomy of distal 9p and 10q might also be associated with an insufficient gene dosage effect. Duplications of the locus DSS can lead to a failure of testicular development and a duplication of the region containing SOX9 has been implicated in XX sex reversal. Transgenic studies in mouse have shown, however, that this mammal is less sensitive to gene dosage than man. Here, we will try to put in place the known pieces of the jigsaw puzzle that is sex determination in mammals, as far as current knowledge obtained from man and animal models allows. We are certain that from this attempt more questions than answers will arise. PMID- 11420126 TI - Transcriptional cascades during spermatogenesis: pivotal role of CREM and ACT. AB - The gene CREM plays key physiological and developmental roles within the hypothalamic--pituitary--gonadal axis. We have previously shown that CREM is highly expressed in male postmeiotic cells. Spermiogenesis is a complex process by which postmeiotic male germ cells differentiate into mature spermatozoa. CREM regulates the expression of a number of post-meiotic genes involved in the process of spermiogenesis. Using homologous recombination we have generated CREM mutant mice that display a complete block at the first step of spermiogenesis. The molecular mechanism by which CREM elicits its regulatory function involves ACT (Activator of CREM in Testis), a testis-specific coactivator constituted by a repeat of four and half LIM domains. ACT is coordinately expressed with CREM, associates with it and confers a powerful transcriptional activation function. It is able to bypass the classical requirement of CREM phosphorylation and recruiting of CBP. PMID- 11420127 TI - Anti-Mullerian hormone and its receptors. AB - Anti-Mullerian hormone (AMH), a member of the transforming growth factor-beta family, is an important factor of male sex differentiation. It is produced by Sertoli cells from the time of fetal sex differentiation to puberty. AMH is also produced by granulosa cells from the time of birth to the end of ovarian activity. As other members of the transforming growth factor-beta family, AMH signals through two related but distinct receptors, both serine/threonine kinases with a single transmembrane domain, called type II and type I. The type II receptor has been cloned in 1994 and is expressed solely in AMH target organs. Engagement of the type I receptor BMPR-IB and downstream effector Smad1 by AMH has recently been demonstrated, however, its role in AMH biological actions remains to be proven. PMID- 11420128 TI - Approaches to define the role of SF-1 at different levels of the hypothalamic pituitary-steroidogenic organ axis. AB - Targeted gene disruption has produced knockout mice lacking the orphan nuclear receptor steroidogenic factor 1 (SF-1). These SF-1 knockout mice lacked adrenal glands and gonads, resulting in adrenocortical insufficiency and sex reversal of their internal and external genitalia. They also had impaired expression of pituitary gonadotropins and agenesis of the ventromedial hypothalamic nucleus (VMH), confirming roles of SF-1 at multiple levels of the hypothalamic-pituitary steroidogenic tissue axis. Using the Cre-loxP system, we now have generated mice in which SF-1 is inactivated selectively in the anterior pituitary. These pituitary-specific SF-1 knockout mice were sterile and failed to exhibit sexual maturation. Histologically, their gonads were markedly hypoplastic, weighing only approximately 5% of the gonads of wild-type mice. Consistent with an important role of SF-1 in gonadotropes, there were no cells in the pituitary gland that expressed either follicle-stimulating hormone (FSH) or luteinizing hormone (LH). These pituitary-specific SF-1 knockout mice are a novel genetic model of hypogonadotropic hypogonadism and establish essential roles of SF-1 in gonadotropin expression. PMID- 11420129 TI - Gonadotropic control of ovarian follicular growth and development. AB - Development-related paracrine cues that sensitize follicles to follicle stimulating hormone (FSH) and luteinizing hormone (LH) are crucial to the emergence of a single dominant follicle in each ovulatory menstrual cycle. Sex steroids, insulin-like growth factors and members of the transforming growth factor-beta superfamily are key players in the follicular paracrine system. FSH acts through membrane-associated granulosa cell receptors (FSHR) to stimulate granulosa cell proliferation and differentiation. The most responsive follicle at the beginning of the cycle is the first to produce estrogen and express granulosa cell LHR. Paracrine signalling activated by FSH and LH sustains growth and oestrogen secretion until an ovulation-inducing LH surge is discharged by the pituitary gland. LH then reprograms granulosa cell function, leading to terminal differentiation (luteinization) rupture of the follicle wall, and release of the fertilizable egg. The genes regulated by the LH surge orchestrate profound changes in sex steroid production, metabolism and action which are necessary for ovulation. Preovulatory granulosa cells also increase their ability to metabolise cortisone to cortisol, which may be part of a local anti-inflammatory mechanism to promote rapid healing of the ruptured ovarian surface. PMID- 11420130 TI - Origin, differentiation and regulation of fetal and adult Leydig cells. PMID- 11420131 TI - Kallmann syndrome: towards molecular pathogenesis. AB - Gonadotropin Releasing Hormone (GnRH) is a key regulator of reproduction and sexual behaviour. During the last decade, embryological studies have clarified the question of the early development of GnRH-synthesising neurones before the onset of neurosecretion. These studies have revealed the existence of a topographical link between GnRH-synthesising neurones and the embryonic olfactory system, thereby shedding new light on Kallmann syndrome, a developmental disease characterised by the association of hypogonadotropic hypogonadism and anosmia (or hyposmia). Although Kallmann syndrome was identified as an inherited disease in the forties, familial cases of the disease are infrequent. However, the identification, by positional cloning strategies, of the gene underlying the X chromosome linked form of the disease (KAL-1) has opened the way to molecular pathophysiology. KAL-1 encodes an extracellular glycoprotein of compound modular structure. The protein, named anosmin-1, has been produced in a transfected mammalian cell line and purified. Polyclonal and monoclonal antibodies have been generated, which allowed us to study the distribution of the protein during the period of human organogenesis (4--10 embryonic weeks), by immunohistofluorescence. During this developmental period, anosmin-1 is a locally restricted component of various extracellular matrices (interstitial matrices and basement membranes). Later in embryonic life, KAL-1 expression apparently becomes restricted to definite neuronal populations. Based on the distribution of anosmin 1 in the early olfactory system, the pathogenesis of the olfactory loss and GnRH deficiency in X-linked Kallmann syndrome is discussed. PMID- 11420132 TI - Inherited disorders of GnRH and gonadotropin receptors. AB - Gonadotropin and GnRH receptors belong to the family of G protein coupled receptors. Gain of function mutations have been described, yielding constitutively active receptors. In the case of the LH receptor these dominant mutations determine familial male limited precocious puberty. Somatic mutations of this receptor may in some cases provoke Leydig-cell adenomas. The constitutive LH receptor is not associated with female precocious puberty. Inactivating mutations are recessive. Alterations in the GnRH receptor determine hypogonadotropic hypogonadism. The clinical diagnosis of this etiology of hypogonadism is extremely difficult, especially in sporadic cases. Mutations of gonadotropin receptors determine primary amenorrhea in girls, whereas in boys they are responsible for Leydig cell aplasia or hypoplasia (LH receptor) or of a variable alteration of spermatogenesis (FSH receptor). Mutations provoking only partial alterations of receptor functions are relatively more frequent, than those inducing complete receptor inactivity. They provide interesting insights into the physiology of GnRH and gonadotropin action. PMID- 11420133 TI - Inherited disorders of the gonadotropin hormones. AB - Pulsatile GnRH acts at the GnRH receptor on gonadotropes to stimulate gonadotropin gene expression, hormone synthesis and secretion. The pituitary gonadotropins, LH and FSH, stimulate steroid production and gametogenesis in males and in females. Gonadotropin production thus requires the normal development and function of hypothalamic GnRH-producing neurons and pituitary gonadotrope cells. Genes involved in gonadotrope development and/or gene expression include SF1, DAX1, KAL, GNRHR, PC1, HESX1, LHX3, PROP1, LH beta, and FSH beta. Given the complex control of gonadotropin biosynthesis and secretion, it is not surprising that genetic abnormalities have been identified at several of these steps. Some of the mutations that will be reviewed include: (1) SF1 and DAX1-orphan nuclear receptors that are expressed at multiple levels throughout the reproductive axis; (2) KAL-X-linked Kallmann syndrome, where there is abnormal development of hypothalamic GnRH-producing neurons; (3) PC1-causing abnormal processing of GnRH and GNRHR mutations that impair action at the GnRH receptor; (4) HESX1, LHX3, PROP1-abnormal development/function of the gonadotrope cell lineage; (5) LH beta and FSH beta-mutations in the gonadotropin genes that cause structural abnormalities in the hormones. Although all of these gene defects lead to gonadotropin deficiency, each disorder is associated with unique phenotypic or hormonal features. Characterization of the molecular basis of gonadotropin deficiency is useful for directing therapy and for genetic counseling. Identification of these mutations also provides insight into the pathways that govern reproduction. PMID- 11420134 TI - Reproductive functions of the progesterone receptor isoforms: lessons from knock out mice. AB - Progesterone plays a central coordinate role in diverse reproductive events associated with establishment and maintenance of pregnancy. In humans and other vertebrates, the biological activities of progesterone are mediated by two proteins, A (PR-A) and B (PR-B) that arise from the same gene and function as progesterone activated transcription factors that exhibit different transcription regulatory activities in vitro. Mice lacking both PR isoforms (PRKO mice) exhibit pleiotropic reproductive abnormalities. To address the physiological role of the individual isoforms, we have selectively ablated PR-A expression in mice (PRAKO). We have demonstrated that PR-B mediates a subset of the reproductive functions of P. Ablation of PR-A does not affect responses of the mammary gland or thymus to P but results in severe abnormalities in ovarian and uterine function. Analysis of urine function of PRAKP mice reveals an unexpected P-dependent proliferative activity of PR-B in the epithelium and provides evidence that the tissue-specific reproductive effects of this isoform are due to specificity of target gene transactivation rather than differences in tissue-specific expression relative to PR-A. Taken together, our data indicate that PR-A and PR-B act in vivo as two functionally distinct transcription factors. PMID- 11420135 TI - Molecular basis of androgen insensitivity. AB - Androgens are important steroid hormones for expression of the male phenotype. They have characteristic roles during male sexual differentiation, during development and maintenance of secondary male characteristics, and during the initiation and maintenance of spermatogenesis. The two most important androgens in this respect are testosterone and 5 alpha-dihydrotestosterone. Each androgen has its own specific role during male sexual differentiation, testosterone is involved in the development and differentiation of Wolffian duct derived structures, whereas 5 alpha-dihydrotestosterone, a metabolite of testosterone, is the active ligand in the urogenital sinus and tubercle and their derived structures. The actions of androgens are mediated by the androgen receptor. This ligand dependent transcription factor belongs to the superfamily of nuclear receptors, including those for the other steroid hormones. The androgen receptor gene is located on the X-chromosome at Xq11--12 and codes for a protein with a molecular mass of approximately 110 kDa. Only one androgen receptor cDNA has been identified sofar, despite two different ligands. It is generally accepted that defects in the androgen receptor gene prevent the normal development of both internal and external male structures in 46, XY individuals. The end-organ resistance to androgens has been designated as androgen insensitivity syndrome (AIS) and is distinct from other forms of male pseudohermaphroditism like 17 beta hydroxy-steroid dehydrogenase type 3 deficiency, leydig cell hypoplasia due to inactivating LH receptor mutations or 5 alpha-reductase type 2 deficiency. Furthermore, two additional pathological situations are associated with abnormal androgen receptor structure and function -- spinal and bulbar muscular atrophy (SBMA, or Kennedy's disease) and prostate cancer. In the AR gene, four different types of mutations have been detected in DNA from individuals with AIS -- (i) single point mutations resulting in amino acid substitutions or premature stopcodons; (ii) nucleotide insertions or deletions most often leading to a frame shift and premature termination; (iii) complete or partial gene deletions; and (iv) intronic mutations in either splice donor or acceptor sites, which affect the splicing of AR RNA. The main phenotypic characteristics of individuals with the complete androgen insensitivity syndrome (CAIS) are, female external genitalia, a short, blind ending vagina, the absence of Wolffian duct derived structures, the absence of a prostate, development of gynecomastia and the absence of pubic and axillary hair. Usually testosterone levels are elevated at the time of puberty, while also elevated LH levels are found. In the partial androgen insensitivity syndrome (PAIS) several different phenotypes are evident, ranging from individuals with predominantly a female appearance to persons with ambiguous genitalia, or individuals with a predominantly male phenotype. At puberty, elevated LH, testosterone and estradiol levels are observed. Individuals with mild symptoms of undervirilization (mild androgen insensitivity syndrome (MAIS)) and infertility have been described as well. Phenotypic variation between individuals in different families has been described for several mutations. However, in cases of CAIS no phenotypic variation has been described within one single family, in contrast to families with individuals with PAIS. In general AIS, can be routinely analyzed and more than 150 different mutations have been reported now. Differential diagnosis of AIS is possible with syndromes presenting with almost similar phenotypes but with a completely different molecular cause. PMID- 11420136 TI - Photodynamic action of actinomycin D: an EPR spin trapping study. AB - Actinomycin D is one of the most widely studied anticancer antibiotic that binds to both double-stranded and single-stranded DNA, and this binding greatly enhances the DNA photosensitization. By use of electron paramagnetic resonance spin trapping techniques, both superoxide radical anion and the radical anion of actinomycin D were identified as important intermediates in the photodynamic process. A mechanism of electron transfer from a DNA base to excited actinomycin D was proposed. These novel findings may shed new light on future application of this drug in photodynamic therapy or cleavage of DNA in unique and controllable ways. PMID- 11420137 TI - Calorimetrically obtained information about the efficiency of ectoine synthesis from glucose in Halomonas elongata. AB - Compatible solutes are becoming more and more attractive commercially. Thus, knowledge of the efficiency of synthesis of compatible solutes from different carbon substrates is very important. As the growth rate and rates of formation of compatible solutes correspond to the heat flux, calorimetric measurements are particularly suitable for providing this information. By growing microorganisms continuously in a calorimeter, and generating a feeding stream with gradually increasing salinity without changing any other growth conditions, we were able to determine the efficiency of growth-associated synthesis of compatible solutes. This was shown for Halomonas elongata DMSZ 2581(T) growing on glucose, which synthesizes (at 25 degrees C) 1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid (ectoine) as its main osmotic counterweight. The requirement of biologically usable energy for its growth-associated synthesis was found to be very low: a 100% efficiency of the conversion of the substrate-carbon into ectoine is both theoretically possible and was reached approximately in practice. The growth rate and yield coefficient were essentially independent of the ectoine formation rate, and the rate of substrate-carbon assimilation was far greater than the rate of dissimilation. The specific maximum growth rate was limited by the rate of formation of ectoine. PMID- 11420138 TI - In situ analysis of mineral content and crystallinity in bone using infrared micro-spectroscopy of the nu(4) PO(4)(3-) vibration. AB - Measurements of bone mineral content and composition in situ provide insight into the chemistry of bone mineral deposition. Infrared (IR) micro-spectroscopy is well suited for this purpose. To date, IR microscopic (including imaging) analyses of bone apatite have centered on the nu(1),nu(3) PO(4)(3-) contour. The nu(4) PO(4)(3-) contour (500-650 cm(-1)), which has been extensively used to monitor the crystallinity of hydroxyapatite in homogenized bone samples, falls in a frequency region below the cutoff of the mercury-cadmium-telluride detectors used in commercial IR microscopes, thereby rendering this vibration inaccessible for imaging studies. The current study reports the first IR micro-spectroscopy spectra of human iliac crest cross sections in the nu(4) PO(4)(3-) spectral regions, obtained with a synchrotron radiation source and a Cu-doped Ge detector coupled to an IR microscope. The acid phosphate (HPO(4)(2-)) content and mineral crystallite perfection (crystallinity) of a human osteon were mapped. To develop spectra-structure correlations, a combination of X-ray powder diffraction data and conventional Fourier transform IR spectra have been obtained from a series of synthetic hydroxyapatite crystals and natural bone powders of various species and ages. X-ray powder diffraction data demonstrate that there is an increase in average crystal size as bone matures, which correlates with an increase in the nu(4) PO(4)(3-) FTIR absorption peak ratio of two peaks (603/563 cm(-1)) within the nu(4) PO(4)(3-) contour. Additionally, the IR results reveal that a band near 540 cm(-1) may be assigned to acid phosphate. This band is present at high concentrations in new bone, and decreases as bone matures. Correlation of the nu(4) PO(4)(3-) contour with the nu(2) CO (3)(2-) contour also reveals that when acid phosphate content is high, type A carbonate content (i.e., carbonate occupying OH(-) sites in the hydroxyapatite lattice) is high. As crystallinity increases and acid phosphate content decreases, carbonate substitution shifts toward occupation of PO(4)(3-) sites in the hydroxyapatite lattice. Thus, IR microscopic analysis of the nu(4) PO(4)(3-) contour provides a straightforward index of both relative mineral crystallinity and acid phosphate concentration that can be applied to in situ IR micro-spectroscopic analysis of bone samples, which are of interest for understanding the chemical mechanisms of bone deposition in normal and pathological states. PMID- 11420139 TI - Diadenosine phosphates and S-adenosylmethionine: novel boron binding biomolecules detected by capillary electrophoresis. AB - There is evidence that boron has a physiological role in animals and humans, but the search for boron binding biomolecules has been difficult because useful radioactive boron isotopes do not exist. To overcome this limitation we used capillary electrophoresis to identify and quantify boron binding to biomolecules by detecting the negative charge boron imparts to ligands. The effect of molecular structure and proximal electronic charges of adenosine and molecules with adenosine moieties including S-adenosylmethionine (SAM) and diadenosine polyphosphates (Ap(n)A) were compared. The boron affinity of the test species varied with the rank order SAM congruent with Ap(6)A congruent with Ap(5)A>Ap(4)A>Ap(3)A congruent with NAD(+)>Ap(2)A>NADH congruent with 5'ATP>5'ADP>5'AMP>adenosine>3'AMP congruent with 2'AMP congruent with cAMP congruent with adenine. Test species with vicinal cis-diols bound boron; species without those moieties did not. Boron binding affinity increased when proximal cationic moieties were present. Anionic moieties remote from the cis-hydroxyl binding site also positively influenced boron binding affinity. In the Ap(n)A species, cooperative complexing of boron between the terminal ribose moieties apparently occurred. In these species boron affinity greater than expected for two monocomplexes was observed and binding affinities increased as more phosphate groups (beyond three) were present separating the terminal moieties. Our results indicate that Ap(6)A, Ap(5)A, Ap(4)A, Ap(3)A, and SAM have higher affinities for boron than any other currently recognized boron ligand present in animal tissues including NAD(+). PMID- 11420140 TI - Mitochondrial NADH-cytochrome b(5) reductase plays a crucial role in the reduction of D-erythroascorbyl free radical in Saccharomyces cerevisiae. AB - The relevance of NADH-cytochrome b(5) reductase to the NADH-dependent reduction of D-erythroascorbyl free radical was investigated in Saccharomyces cerevisiae. MCR1, which is known to encode NADH-cytochrome b(5) reductase in S. cerevisiae, was disrupted by the insertion of URA3 gene into the gene of MCR1. In the mcr1 disruptant cells, the activity of NADH-D-erythroascorbyl free radical reductase almost disappeared and the intracellular level of D-erythroascorbic acid was about 11% of that of the congenic wild-type strain. In the transformant cells carrying MCR1 in multicopy plasmid, the intracellular level of D-erythroascorbic acid and the activity of NADH-D-erythroascorbyl free radical reductase increased up to 1.7-fold and 2.1-fold, respectively. Therefore, it indicated that the MCR1 product, mitochondrial NADH-cytochrome b(5) reductase, plays a key role in the NADH-dependent reduction of D-erythroascorbyl free radical in S. cerevisiae. On the other hand, the mcr1 disruptant cells were hypersensitive to hydrogen peroxide and menadione, and overexpression of MCR1 made the cells more resistant against oxidative stress. These results suggested that the mitochondrial NADH cytochrome b(5) reductase functions as NADH-D-erythroascorbyl free radical reductase and plays an important role in the response to oxidative damage in S. cerevisiae. PMID- 11420141 TI - Biological activities of human mannose-binding lectin bound to two different ligand sugar structures, Lewis A and Lewis B antigens and high-mannose type oligosaccharides. AB - The biological activities of mannose-binding lectin (MBL) which binds to different ligands on mammalian cells were examined using two types of Colo205 cells, a human colon adenocarcinoma cell line: one naturally expressing Lewis A and Lewis B antigens as ligands for MBL (NT-Colo205), and the other modified to express high-mannose type oligosaccharides by treatment with benzyl-2-acetamide-2 deoxy-alpha-galactopyranoside and 1-deoxymannojirimycin (Bz+dMM-Colo205). Although the final lysis was not observed, the deposition of C4 and C3 was observed on both types of Colo205 cells after treatment with MBL and complements as a result of complement activation by MBL. MBL bound to Bz+dMM-Colo205 could also activate human peripheral blood leukocytes and induce superoxide production; however, MBL bound to NT-Colo205 could not. This may be explained by the lower affinity of MBL to Lewis A and Lewis B antigens than to high-mannose type oligosaccharides under physiological conditions, since MBL bound to NT-Colo205 was more easily released from the cell surface than that bound to Bz+dMM-Colo205 at 37 degrees C. These findings suggest that the difference in the affinity of MBL to its ligands could influence the expression of some biological activities of MBL. PMID- 11420142 TI - Purification, characterization and developmental expression of rat brain PSP protein. AB - We have isolated a perchloric acid-soluble protein designated as B-PSP1 from the postmitochondria supernatant fraction of rat brain. It was purified by gel filtration and anti-PSP1 affinity chromatography. Immunoblotting, peptide mapping, partial amino acid sequencing and reverse transcriptase-polymerase chain reaction showed that the amino acid sequence of B-PSP1 was identical with that of PSP isolated from rat liver. B-PSP1 was expressed in all regions including frontal cortex, posterior cortex, cerebellum, hippocampus, olfactory bulb, striatum, thalamus, midbrain, entorhinal cortex, pons, medulla and spinal cord. Immunohistochemical study showed that the expression of B-PSP1 was observed in ependymal cells of choroid plexus and glial cells of the other region. The expression of B-PSP1 in brain increased gradually from the first day to the 60th day of postnatal age, but the expression of B-PSP1 was slower than that of GFAP which is a marker protein of glial cells. The expression of PSP may be related to the cellular function rather than the developmental regulation of the glial cells. Thus, our findings are the first report on the presence of a PSP in rat brain which may be involved in the regulation of cellular function. PMID- 11420143 TI - Epitopes in the interacting regions of beta-dystroglycan (PPxY motif) and dystrophin (WW domain). AB - The dystroglycan gene produces two products from a single mRNA, the extracellular alpha-dystroglycan and the transmembrane beta-dystroglycan. The Duchenne muscular dystrophy protein, dystrophin, associates with the muscle membrane via beta dystroglycan, the WW domain of dystrophin interacting with a PPxY motif in beta dystroglycan. A panel of four monoclonal antibodies (MANDAG1-4) was produced using the last 16 amino acids of beta-dystroglycan as immunogen. The mAbs recognized a 43 kDa band on Western blots of all cells and tissues tested and stained the sarcolemma in immunohistochemistry of skeletal muscle over a wide range of animal species. A monoclonal antibody (mAb) against the WW domain of dystrophin, MANHINGE4A, produced using a 16-mer synthetic peptide, recognized dystrophin on Western blots and also stained the sarcolemma. We have identified the precise sequences recognized by the mAbs using a phage-displayed random 15 mer peptide library. A 7-amino-acid consensus sequence SPPPYVP involved in binding all four beta-dystroglycan mAbs was identified by sequencing 17 different peptides selected from the library. PPY were the most important residues for three mAbs, but PxxVP were essential residues for a fourth mAb, MANDAG2. By sequencing five different random peptides from the library, the epitope on dystrophin recognized by mAb MANHINGE4A was identified as PWxRA in the first beta strand of the WW domain, with the W and R residues invariably present. A recent three-dimensional structure confirms that the two epitopes are adjacent in the dystrophin-dystroglycan complex, highlighting the question of how the two interacting motifs can also be accessible to antibodies during immunolocalization in situ. PMID- 11420144 TI - Oxazole yellow homodimer YOYO-1-labeled DNA: a fluorescent complex that can be used to assess structural changes in DNA following formation and cellular delivery of cationic lipid DNA complexes. AB - To improve transfection efficiency following delivery of plasmid expression vectors using lipid-based carriers, it is crucial to define structural characteristics of the lipid/DNA complexes that optimize transgene expression. Due to its strong affinity for DNA and high quantum yield, the fluorescent DNA intercalator YOYO-1 was used as a tool to assess changes in DNA that occur following lipid binding and cell delivery. In this study, the stability of the dye/DNA complex following binding of poly-L-lysine or monocationic lipids is characterized. More than 98% of the fluorescence measured for a defined DNA/YOYO 1 complex was lost when DNA was condensed using poly-L-lysine. This loss in fluorescence could be attributed to displacement of bound dye. In contrast, more than 30% of the fluorescence of the dye-labeled DNA was retained after formation of cationic lipid/DNA complexes. Significantly, the results illustrate differences in structural changes cationic lipids and PLL exert on plasmid DNA. The fluorescent lipid/DNA complex was used to assess DNA delivery to murine B16/BL6 cells in vitro. An assay relying on fluorescence resonance energy transfer between bound YOYO-1 and propidium iodide was used to distinguish between DNA attached to the cell surface and internalized DNA. PMID- 11420145 TI - (CGA)(4): parallel, anti-parallel, right-handed and left-handed homoduplexes of a trinucleotide repeat DNA. AB - Conformational properties of microsatellite DNA regions are the probable reason of their expansions in genomes which lead to serious genetic diseases in some cases. Using CD spectroscopy, UV absorption spectroscopy and polyacrylamide gel electrophoresis, we study in this paper conformational properties of (CGA)(4) and compare them with those of (CAG)(4) - a related repeat, connected with Huntington's disease. We show that (CGA)(4) can adopt several distinct conformations in solution. Around neutral pH it forms a parallel-stranded homoduplex containing C(+).C, G.G, and A.A base pairs. Under the same conditions (CAG)(4) forms a hairpin. At slightly alkaline pH values and low ionic strength, (CGA)(4) also folded into a hairpin which transformed into a bimolecular anti parallel homoduplex at increasing salt concentrations. The duplex easily isomerized into left-handed Z-DNA, implying that the mismatched adenines between G.C pairs facilitate rather than hinder the B-Z transition. No similar changes took place with (CAG)(4). Thus, the conformational repertoire of (CGA)(4) includes parallel, anti-parallel, right-handed, and left-handed homoduplexes. In contrast, (CAG)(4) invariably adopts only a single conformation, namely the very stable hairpin. PMID- 11420146 TI - Reduction of organic nitrates catalysed by xanthine oxidoreductase under anaerobic conditions. AB - Xanthine oxidoreductase catalyses the anaerobic reduction of glyceryl trinitrate (GTN), isosorbide dinitrate and isosorbide mononitrate to inorganic nitrite using xanthine or NADH as reducing substrates. Reduction rates are much faster with xanthine as reducing substrate than with NADH. In the presence of xanthine, urate is produced in essentially 1:1 stoichiometric ratio with inorganic nitrite, further reduction of which is relatively slow. Organic nitrates were shown to interact with the FAD site of the enzyme. In the course of reduction of GTN, xanthine oxidoreductase was progressively inactivated by conversion to its desulpho form. It is proposed that xanthine oxidoreductase is one of several flavoenzymes that catalyse the conversion of organic nitrate to inorganic nitrite in vivo. Evidence for its further involvement in reduction of the resulting nitrite to nitric oxide is discussed. PMID- 11420147 TI - Cloning and expression of cDNAs encoding alpha1,3-fucosyltransferase homologues from Arabidopsis thaliana. AB - The core alpha1,3-fucosyltransferases are involved in the synthesis of glycans specific to plants and invertebrates which are known to be immunogenic and allergenic. We report the identification, isolation and characterisation of the cDNAs of three genes (FucTA, FucTB and FucTC) encoding proteins similar to alpha1,3-fucosyltransferases in Arabidopsis thaliana. Reverse transcription polymerase chain reaction was used to amplify the full length coding sequence of FucTA. The FucTA gene, which consists of seven exons, encodes a presumptive protein of 501 amino acids showing an overall sequence identity of 66% to the protein encoded by the recently isolated mung bean Fuc-T C3 cDNA. FucTA was expressed in Pichia pastoris under the control of the AOX1 gene promoter. The soluble enzyme was found to catalyse the same reaction as mung bean core alpha1,3 fucosyltransferase as judged by analyses of the products by MALDI-TOF and high performance liquid chromatography. The FucTB cDNA was isolated from a lambda-ZAP library, but the clone used an alternative splicing site between the second and third exon resulting in a premature stop codon. The FucTC gene encodes a protein with less than 40% identity to FucTA across 115 amino acids of a total of 401 amino acids and is a member of a new sub-family of plant alpha1,3/4 fucosyltransferase homologues. PMID- 11420148 TI - Multiple changes in gene expression in chronic human Achilles tendinopathy. AB - Atlas cDNA cell interaction arrays (CLONTECH) were used to examine degenerate tissue from four patients with Achilles tendon disorders, in order to identify changes in expression of genes important in cell-cell and cell-matrix interactions. The greatest difference between normal (post-mortem) and degenerate tissue samples was in the level of MMP-3 (stromelysin) mRNA, which was down regulated in all the degenerate samples. Quantitative RT-PCR assay of RNA extracted from paired 'normal' and degenerate Achilles tendon tissue samples taken from tendons during surgery mirrored the results of the arrays. Levels of MMP-3 mRNA were lower, whereas levels of type-I and type-III collagen mRNAs were significantly higher, in the degenerate compared to the 'normal' samples. Immunoblotting of proteins extracted from the same tendon samples showed that three of four normal tissue samples taken from individuals without apparent tendon disorder had much higher levels of MMP-3 protein than 'normal' or degenerate samples from patients with tendinosis. We suggest that MMP-3 may play an important role in the regulation of tendon extracellular matrix degradation and tissue remodelling. PMID- 11420149 TI - Specific RGTA increases collagen V expression by cultured aortic smooth muscle cells via activation and protection of transforming growth factor-beta1. AB - Regenerating agents (RGTA) are defined as heparan sulfate mimics, which in vivo stimulate tissue repair. RGTA are obtained by controlled grafting of carboxymethyl and sulfate groups on dextran polymers. RGTA are selected in vitro, on their ability to protect heparin binding growth factors such as TGF-beta1 for example, as well as to alter extracellular matrix biosynthesis. We had reported that RGTA were able to modulate smooth muscle cell (SMC) collagen biosynthesis. Here, we demonstrated that a specific RGTA (RG-1503), altered differentially collagen type expression by post-confluent SMC and that this action involves TGF beta1. RG-1503 decreased, by 50%, collagen I and III biosynthesis and stimulated specifically, by twofold, collagen V biosynthesis. TGF-beta1 stimulated collagen I and V by 1.5- and threefold, respectively. A synergic action for RGTA in association with TGF-beta1 was observed specifically for collagen V expression (eightfold increase). The stimulation of collagen V biosynthesis by RGTA was abolished by TGF-beta1 neutralizing antibodies. These modulations occurred at protein and mRNA levels. RG-1503 did not alter TGF-beta1 mRNA steady state level or total TGF-beta1 protein content (latent+active forms). However, RG-1503 significantly induced an elevated proportion of active TGF-beta1 form, which could result from the selective protection from proteolytic degradation of TGF beta1 by RG-1503. These data open a rationale for understanding the stimulation of tissue repair induced by RGTA, and also, a new insight for developing drugs adapted to inhibit excess collagen deposition in smooth muscle cells associated vascular disorder, and in fibrotic diseases. PMID- 11420150 TI - Fibroblasts require protein kinase C activation to respond to hyaluronan with increased locomotion. AB - Hyaluronan (HA) stimulates the motility of some but not all cell types. Here, we show that HA-promoted random motility of ras-transformed 10T1/2 (C3) fibroblasts requires activation of protein kinase C and is associated with rapid uptake of HA in a CD44 and RHAMM-dependent manner. The addition of HA to parental 10T1/2 fibroblasts (parental cells) does not stimulate random motility, but these cells can be 'primed' to respond to HA by treatment with the phorbol ester, PMA, for 4 6 h. This effect of PMA requires protein synthesis, PKC activity and is associated with enhanced uptake of HA. These results suggest that the ability of cells to respond to HA is regulated by a protein kinase C-dependent process that may promote uptake of HA. PMID- 11420151 TI - Gelatinase A (MMP-2) activation by skin fibroblasts: dependence on MT1-MMP expression and fibrillar collagen form. AB - The respective requirements of collagen and MT1-MMP in the activation of MMP-2 by primary fibroblast cultures were explored further. Three-dimensional gels enriched in human collagen types I and III or composed of recombinant human type II or III collagen, caused increased MT1-MMP production (mRNA and protein) and induced MMP-2 activation. Only marginal induction was seen with dried monomeric collagen confirming the need for collagen fibrillar organisation for activation. To our surprise, relatively low amounts (as low as 25 microg/ml) of acid soluble type I collagen added to fibroblast cultures also induced potent MMP-2 activation. However, the requirement for collagen fibril formation by the added collagen was indicated by the inhibition seen when the collagen was pre-incubated with a fibril-blocking peptide, and the reduced activation seen with alkali treated collagen preparations known to have impaired fibrilisation. Pre-treatment of the collagen with sodium periodate also abrogated MMP-2 activation induction. Further evidence of the requirement for collagen fibril formation was provided by the lack of activation when type IV collagen, which does not form collagen fibrils, was added in the cultures. Fibroblasts derived from MT1-MMP-deficient mice were unable to activate MMP-2 in response to either three-dimensional collagen gel or added collagen solutions, compared to their littermate controls. Collectively, these data indicate that the fibrillar structure of collagen and MT1-MMP are essential for the MMP-2 activational response in fibroblasts. PMID- 11420152 TI - Role of the subchondral vascular system in endochondral ossification: endothelial cell-derived proteinases derepress late cartilage differentiation in vitro. AB - Endochondral ossification in growth plates proceeds through several consecutive steps of late cartilage differentiation leading to chondrocyte hypertrophy, vascular invasion, and, eventually, to replacement of the tissue by bone. The subchondral vascular system is essential for this process and late chondrocyte differentiation is subject to negative control at several checkpoints. Endothelial cells of subchondral blood vessels not only are the source of vascular invasion accompanying the transition of hypertrophic cartilage to bone but also produce factors overruling autocrine barriers against late chondrocyte differentiation. Here, we have determined that the action of proteases secreted by endothelial cells were sufficient to derepress the production of the hypertrophy-markers collagen X and alkaline phosphatase in arrested populations of chicken chondrocytes. Signalling by thyroid hormones was also necessary but endothelial factors other than proteinases were not. Negative signalling by PTH/PTHrP- or TGF-beta-receptors remained unaffected by the endothelial proteases whereas signalling by FGF-2 did not suppress, but rather activated late chondrocyte differentiation under these conditions. A finely tuned balance between chondrocyte-derived signals repressing cartilage maturation and endothelial signals promoting late differentiation of chondrocytes is essential for normal endochondral ossification during development, growth, and repair of bone. A dysregulation of this balance in permanent joint cartilage also may be responsible for the initiation of pathological cartilage degeneration in joint diseases. PMID- 11420153 TI - Public awareness of aortic aneurysm. PMID- 11420154 TI - The human heart: from the heart surgeon's perspective. PMID- 11420155 TI - Expression of apoptosis-related proteins and structural features of cell death in explanted aortocoronary saphenous vein bypass grafts. AB - This study aimed to investigate the features of cell death occurring in aortocoronary saphenous vein bypass grafts. Human aortocoronary saphenous vein bypass grafts with angiographic luminal stenosis of > 75% were explanted from 14 patients at redo coronary artery bypass grafting. Proteins associated with apoptotic pathways were identified immunohistochemically using antibodies to Bcl 2, Fas, BAX, p53 and CPP32. Cells undergoing DNA fragmentation were identified by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). DNA synthesis was investigated using the antibody to proliferating cell nuclear antigen (PCNA). Ultrastructural features of cell death were examined by electron microscopy. Anti-apoptotic (Bcl-2) and pro-apoptotic (Bax, p53, CPP32 and Fas) proteins were expressed throughout the graft wall, but marked differences in the characteristics of cell death were noted between atherosclerotic and non atherosclerotic areas of the intima. In atherosclerotic areas, pro-apoptotic proteins were widely expressed, but ultrastructural analysis failed to identify cells showing typical features of apoptosis. In these areas, necrotic cells were frequently observed, with negative correlation of Bcl-2 expression with TUNEL. Pro-apoptotic proteins showed no correlation with TUNEL. In contrast, in non atherosclerotic areas of vein grafts, the expression of both anti-apoptotic (Bcl 2) and pro-apoptotic proteins (p53, Bax and CPP32) correlated with TUNEL. In atherosclerotic areas, non-atherosclerotic intimal areas, and in the underlying media, the numbers of TUNEL+ cells correlated with PCNA positivity. Ultrastructurally, apoptotic bodies and features of necrosis were observed in non atherosclerotic areas of grafts. The present observations indicate that in atherosclerotic areas, cell death occurs mainly by necrosis, while in non atherosclerotic areas, cell death occurs by both necrosis and apoptosis. An imbalance between DNA fragmentation and DNA synthesis may contribute to graft instability and failure. PMID- 11420156 TI - CD40 co-stimulatory molecule expression by dendritic cells in primary atherosclerotic lesions in carotid arteries and in stenotic saphenous vein coronary artery grafts. AB - We have previously identified dendritic cells in the intima of human large arteries and stenotic vein coronary bypass grafts. The mechanisms by which these dendritic cells might regulate immune responses in atherosclerotic lesions and stenotic vein grafts are unknown. The aim of the present study was to investigate whether dendritic cells in carotid plaques and in stenotic aortocoronary vein grafts express an immunoregulatory molecule CD40. Segments of wall from eight carotid arteries and three stenotic aortocoronary saphenous vein grafts were obtained at operation. CD40+ cells were detected in all specimens of both occluded and stenotic grafts and carotid plaques. Although CD40+ cells of various cell types were intermingled in most areas within the plaques and stenotic grafts, there were sites where CD40+ cells were located in small groups. Consecutive sections demonstrated that a small population of CD40+ cells stained positively for S100. These CD40+/S100+ cells were clustered within the intima but were also found in the media and adventitia. This suggests that dendritic cells, which accumulate within vessels affected by atherosclerosis and graft disease, express CD40 co-stimulatory molecule. The expression of CD40 molecule on the dendritic cells may be important in regulating T cell responses within atherosclerotic plaques and stenotic vein grafts. PMID- 11420157 TI - Carotid endarterectomy in awake patients with contralateral carotid artery occlusion. AB - OBJECTIVES: Patients with severe stenosis of an internal carotid artery with contralateral occlusion (ICO) are at an increased risk for stroke, and therefore surgical treatment is usually recommended. Carotid endarterectomy (CEA) under regional anesthesia enables constant monitoring of neurologic status and selective shunting in cases of clinically evident cerebral ischemia. In this study, we assess the selective use of shunts based solely on changes in neurological status in awake patients with ICO undergoing CEA as well as their complication rates. METHODS: During 1996-1998, we studied intraoperative findings and results of CEA under regional anesthesia with clinical monitoring of neurological status in two groups: (1) patients with stenosis (> 70% by NASCET) and contralateral occlusion (n = 50) and (2) patients with stenosis and no contralateral occlusion (n = 94). RESULTS: Shunt insertion was required in 42% of group 1, and 6% in group 2. All of the patients in group 1 requiring shunts had stump pressures < 50 torr. The average stump pressure of group 1(40 torr) was significantly lower than that of group 2 (75 torr), and was also lower than that of patients with severe contralateral stenosis (35 patients, 76 torr). Perioperative stroke rates were identical in both groups (2.1%). CONCLUSION: Since ICO patients are at a high risk for brain ischemia during ICA clamping, they require shunt insertion frequently. Patients with no contralateral occlusion require shunting at a much lower rate - even in the presence of severe contralateral stenosis. Regional anesthesia allows for early detection of brain ischemia and therefore, the perioperative results in both groups are similar. PMID- 11420158 TI - Combined carotid endarterectomy and coronary bypass: a decade experience at UCLA. AB - PURPOSE: The purpose of this review was to determine outcomes for combined carotid endarterectomy (CEA) and coronary revascularization (CABG) in patients with asymptomatic carotid stenosis. METHODS: We reviewed the medical records of consecutive combined procedures (CEA and CABG), performed at UCLA Medical Center from October, 1989 to January, 1999. FINDINGS: There were 43 patients, 27 men and 16 women, with a mean age of 71 yr (range 51-87). Thirty-four patients 79% (34/43) had asymptomatic carotid stenosis. Stroke occurred in three patients (3/43 = 6.9%). Stroke ipsilateral to the CEA occurred in two patients: one asymptomatic (1/34 = 2.9%) and one symptomatic (1/9 = 11.1%). CONCLUSIONS: The majority of patients undergoing combined CEA/CABG have asymptomatic carotid stenosis identified in preparation for elective CABG. The asymptomatic carotid subset stroke rate of 2.9% resulting from a combined CEA/CABG is higher than our reported rate for CEA performed alone. In patients with asymptomatic carotid stenosis, the combined procedure should be selectively performed. PMID- 11420159 TI - Metabolic changes in the ischemic penumbra after carotid endarterectomy in stroke patients by localized in vivo proton magnetic resonance spectroscopy (1H-MRS). AB - Carotid endarterectomy (CEA) is known to be effective in reducing recurrent ischemic attacks, sometimes accompanied with the functional improvement, for patients with internal carotid artery (ICA) flow lesions by increase in perfusion and/or removal of embolic sources. However, the exact mechanism of how the CEA affects the cerebral metabolism in relations to the perfusion increase in noninfarcted hypoperfused peripheral areas to the center of the lesion (e.g., ischemic penumbra or border zone) is not yet clearly known. The existence of the ischemic penumbra and its long-term viability has also been argued. We designed a prospective study to investigate the metabolic changes in the ischemic penumbra for patients with ICA flow lesions and cerebral infarct (or ischemia) before and after CEA using localized in vivo proton magnetic resonance spectroscopy ((1)H MRS). The results of (1)H-MRS showed significantly decreased choline (Cho)/creatine (Cr) and increased N-acetylaspartate (NAA)/Cho ratios in the periphery of the lesion for the patients after CEA as compared to those who underwent only medical treatments. The more significant changes in the cerebral metabolite levels were observed in the patients who showed the improved cerebral perfusion by single photon emission computed tomography after CEA than in those who did not. In conclusion, our data suggest the existence of the ischemic penumbra, which were viable for a longer period than previously thought; CEA seems to improve the cerebral metabolism that may result from the improved perfusion at the ischemic penumbra. PMID- 11420160 TI - Femoro-popliteal reconstructions: 'in situ' versus 'reversed' technique. AB - This study examined 191 patients with 'reversed' and 99 patients with 'in situ' femoro-popliteal bypass technique. There were 85 diabetic patients (44.5%) in the group with 'reversed' bypass, and 43 patients (43.43%) in the 'in situ' group. There were 152 (79.68%) smokers in the 'reversed' bypass group, and 80 (80.8%) in the 'in situ' group. The graft patency was confirmed immediately after operation using CW Doppler and then followed up after 1, 6, l2 months and annually thereafter. The statistical analysis was performed using Pearsons chi-square test, Fischer's test and 'Life table' statistic methods. The patients were followed from 3 to 10 yr after surgery. 'In situ' bypass showed better patency than the 'reversed' bypass technique but only in the second and tenth follow-up year (P < 0.05). Also, 'in situ' bypass proved to be better than 'reversed' only in patients with one patent crural artery (P < 0.01). Diabetes and preoperative smoking did not significantly affect late patency regarding this technique (P > 0.05). However, continuous smoking after the operation significantly decreased late patency rate in both groups of patients (P < 0.01). There was no significant difference in the early thrombectomy rate between groups with 'reversed' and 'in situ' bypasses (P > 0.05). The early thrombectomy, however, significantly reduced late patency rate in both groups (P < 0.01). Therefore we suggest 'in situ' bypass in cases with poor run off, small-calibre vein and 'long' bypass. Also, we consider important more frequent physical and Doppler ultrasonographic control in patients who had early thrombectomy. PMID- 11420161 TI - Effect of ischaemic preconditioning, cardiopulmonary bypass and myocardial ischaemic/reperfusion on free radical generation in CABG patients. AB - OBJECTIVE: To investigate the free radicals (FR) generation after ischaemic preconditioning and cardiopulmonary bypass and during reperfusion in CABG patients, and the role of ischaemic preconditioning. METHODS: Forty-three CABG patients were randomised into an ischaemic preconditioning and a control group. The protocol for ischaemic preconditioning was two cycles of 2-min ischaemia followed by 3-min reperfusion. Free radicals were measured using electron spin resonance spectroscopy. Global and right heart functions were collected. RESULTS: The free radicals generation in coronary sinus blood in the ischaemic preconditioning group was 9.7 and 16.6% after the ischaemic preconditioning protocol and 10 min after declamping, 6.8 and 13.3% in the controls. The free radicals in arterial samples were, respectively, 21, 14, 10 and 9% at 10 min, 1, 2 and 24 h after reperfusion. Cardiac index (CI) and right ventricular ejection fraction (RVEF) were improved by ischaemic preconditioning. CONCLUSION: Both ischaemic preconditioning and cardiopulmonary bypass induced free radicals generation. Although ischaemic preconditioning had no effect on free radicals generation after the operation, it protected against postoperative stunning. PMID- 11420162 TI - Tricuspid regurgitation secondary to mitral valve disease: tricuspid annulus function as guide to tricuspid valve repair. AB - METHODS: A prospective analysis was performed on 50 patients (pts) with rheumatic mitral disease and associate secondary tricuspid insufficiency who underwent mitral valve replacement from January 1995 to December 1998. Surgical indication to tricuspid annuloplasty was considered in patients with echocardiographic tricuspid annulus diameter > 21 mm/m2, regardless semiquantitative evaluation of tricuspid insufficiency. De Vega annuloplasty was performed in 33 out of 50 patients. RESULTS: Hospital mortality was 2.0% (CL 0.3-3.6). The follow up of the discharged patients ranged from 3 to 48 months (mean 25 +/- 15.9). Three late deaths occurred (6.1% CL 2.8-9.2). Forty-two patients out of the 46 followed up (91.3% CL 84.9-93.8) were in I or II NYHA class. In eight patients (16.3% of discharged patients) the obtained result has been considered as 'negative late results': persisting moderate (three cases) or moderate-severe (five cases) TrI, together with congestive heart failure requiring a furosemide intake of > 25 mg/day. No patients had severe TrI at follow up. The statistics analysis demonstrated the 'preoperative fraction shortening of the tricuspid annulus' (P = 0.038) as factor predictive of late negative result. The incidence of late negative result was 57.1% among patients with fractional shortening lower than 25% and 0% among those patients with fractional shortening greater than 25% (P = 0.0001). CONCLUSIONS: The choice to treat the tricuspid insufficiency according to indexed tricuspid annulus dimension (> 21 mm/m2) has been effective in terms of clinical efficacy and of late functional result. Fractional shortening of the tricuspid annulus, expression of right ventricular cardiomyopathy in patients with poorest prognosis, affects the postoperative evolution of tricuspid insufficiency. PMID- 11420163 TI - Effect of radial artery harvesting on tissue perfusion and function of the hand. AB - This study was designed to measure the degree to which hand and forearm blood flow is reduced following harvesting of the radial artery in myocardial revascularization surgery and determine whether there is an effect on hand function. Twenty patients who had the radial artery used for myocardial revascularization underwent bilateral blood flow assessment of hands and forearms using Technetium-99m human serum albumin and clinical evaluation of hand function. Mean tissue perfusion in ml/100 ml/min +/- SD was as follows: donor hand 21.9 +/- 5.6, non-donor hand 25.5 +/- 6.1 (P = 0.00043), donor arm (hand and forearm) 17.5 +/- 3.7 and non-donor arm (hand and forearm) 21 +/- 5.1 (P = 0.000681). No clinical evidence of hand claudication was detected. This study suggests that removal of the radial artery reduces the tissue perfusion of the hand and forearm but does not affect hand function in the short term. The use of radial artery grafts in patients at risk of developing peripheral vascular disease should be carefully considered PMID- 11420164 TI - Effective surgical management of high-risk coronary patients using preoperative intra-aortic balloon counterpulsation therapy. AB - The proportion of high-risk coronary patients submitted to surgical myocardial revascularization (CABG) is steadily increasing. High-risk patients utilize more hospital resources and have a higher procedural cost than low-moderate risk CABG patients. An efficient management is essential to improve outcome and reduce costs. This report entails three study periods. In an initial retrospective study coronary high-risk criteria were established. At least two of the following factors were required: redo CABG, unstable angina, left main stem stenosis greater than 70%, preoperative left ventricular ejection fraction < 0.30 and diffuse coronary artery disease. Poor preoperative cardiac performance was the major contributing factor for poor outcome. Intra-aortic balloon counterpulsation therapy (IABP) was introduced as preoperative therapy. During a second study period prospective randomized studies found preoperative IABP-therapy efficient, significantly improving both preoperative cardiac index (P < 0.0001), decreasing postoperative mortality (P < 0.0001) and morbidity, shorten intensive care unit stay as well as total hospital stay (P < 0.0001). Drug consumption was significantly reduced (P < 0.0001). Optimal timing was found to be 2 h prior to aortic cross-clamping and the therapy was found highly cost-beneficial with an average 36% reduction of the total procedural cost. During a third study period, well beyond any study protocol period, preoperative IABP therapy was again found highly effective with a close to 100% utilization rate in high-risk patients and continuous efficacy with excellent outcome, despite acceptance of sicker patients. During this post-study evaluation period 1/3 of the high-risk patients presented with 4 of the established risk factors. The use of preoperative IABP therapy is therefore highly recommended for high-risk coronary patients undergoing CABG. PMID- 11420165 TI - Improved reliability of post-operative ventricular pacing by use of bipolar temporary pacing leads. AB - The study compared the clinical reliability of using a bipolar epicardial wire (6495, Medtronic) over a unipolar type (FEP15, Ethicon) for post-operative pacing in coronary artery surgery. Atrial and ventricular wires of both types were implanted in 18 patients. Sensitivities and pacing thresholds were tested for 5 consecutive days. Results show that pacing thresholds were better maintained with the bipolar wire in both atria and ventricles. However, sensing failures were frequent in the atrial position (34% vs 9.3% compared with unipolar). By contrast, in the ventricle, no sensing failures occurred (0% vs 17.6% compared with unipolar). Furthermore, sensing magnitude was significantly better (11.13+/ 1.32 vs 5.65+/-0.53 mV, P<0.001). We conclude that a single 6495 bipolar wire is effective for temporary ventricular pacing, whilst double unipolar wires remain a useful strategy for securing atrial sensing and pacing. PMID- 11420166 TI - Aortic arch and proximal supraaortic arterial repair under continuous antegrade cerebral perfusion and moderate hypothermia. AB - PURPOSE: In this prospective study the clinical and neurological outcome of continuous antegrade cerebral perfusion (ACP) and moderate hypothermia was evaluated in patients undergoing ascending and aortic arch repair including reconstruction of the proximal supraaortic arteries. METHODS: In 50 consecutive patients (mean age 47 yr, range 22-70) aortic arch and supraaortic arterial repair was performed: ascending aorta and aortic arch (n=34) and aortic arch and Bentall procedure (n = 16). In 12 patients the distal anastomosis was performed using the elephant trunk technique. Test-clamping of the innominate artery for 3 min was performed under EEG-monitoring followed by the same procedure for the left carotid artery. Cardiopulmonary bypass was instituted and the innominate artery replaced by a polyester graft before antegrade perfusion was carried out through the graft. While cooling to 28-30 degrees C, the left carotid artery was similarly treated with subsequent antegrade cerebral perfusion. The distal anastomosis was made at or beyond the left subclavian artery under circulatory arrest. During rewarming the innominate and carotid polyester grafts as well as the subclavian artery were anastomosed to the main graft, while antegrade cerebral perfusion was continued. RESULTS: In 46 patients antegrade cerebral perfusion was achieved with a mean volume flow of 12 ml/kg/min and a mean arterial pressure of 54 mmHg. EEG-monitoring delineated stable and symmetrical recordings. In four patients antegrade flow (mean 15 ml/kg/min) and pressure (mean 65 mmHg) had to be increased to establish baseline EEG-recordings. The mean time of circulatory arrest was 18 min. The overall hospital mortality was 6%: two patients died from cerebral infarction and one patient suffered from a ruptured abdominal aortic aneurysm. Three patients (6%) developed a temporary neurological deficit which resolved spontaneously. Two patients (4%) developed renal failure requiring temporary hemodialysis. Pulmonary complications occurred in 12 patients (25%). CONCLUSION: Continuous antegrade cerebral perfusion via selective grafts to the innominate and carotid arteries offers adequate protection in patients undergoing replacement of the ascending aorta or aortic arch and great vessels. This technique allows radical repair and optimal vascular reconstruction without time restrains and avoids the necessity for profound hypothermia PMID- 11420167 TI - Aprotinin reduces the IL-8 after coronary artery bypass grafting. AB - The effect of aprotinin, a protease inhibitor, on myocardial interleukin-8 (IL-8) production after ischemia-reperfusion injury was investigated. Twenty patients who had elective coronary artery bypass grafting were included in this study. Patients were randomly divided into two groups (n = 10 in each). Group A patients received high dose aprotinin (20,000 IU/kg as pretreatment followed by 7500 IU/kg for 6 h) and Group B patients received normal saline as a control. Serum IL-8 levels after the termination of cardiopulmonary bypass (CPB) showed a significant improvement in aprotinin treated group compared to control group (70 +/- 42.6 vs 360.71 +/- 87.9 ng/ml) (P < 0.005). Levels were also significantly higher at post operative 24th hour in patients who did not received aprotinin (340.16 +/- 92.10 vs 96.13 +/- 34.33 ng/ml). However at post-operative 48th hour levels were again higher in control (untreated) group, but the difference was not statistically significant (78.8 +/- 34.4 vs 42.8 +/- 9.29 ng/ml). Aprotinin prevented the IL-8 release from myocytes in ischemia-reperfusion injury. The mechanism is highly dependent on anti-protease activity of aprotinin. PMID- 11420168 TI - Coronary artery stabilization causes endothelial damage: an electron microscopic study on dogs. AB - In recent years, to protect patients from adverse effects of cardiopulmonary bypass, off-pump coronary artery bypass operations are performed. In these operations, effective coronary artery immobilization is the standard for a fine anastomosis. This is usually maintained with the help of prolene immobilization sutures, bulldog occluders or mechanical epicardial devices. The aim of this study is to show whether these stabilization devices cause any damage to the coronary artery endothelium. Sixteen dogs are included in this study and are divided into four groups. The control group consisted of one dog (Group I) and different immobilization techniques are applied to the other groups, which were five dogs each (Groups II, III, and IV). Coronary artery biopsies are obtained, then the samples are studied under scanning electron microscope (SEM) for endothelial injury and all samples are scored. Among the techniques used, bulldog occluders caused less injury on the coronary endothelium than the prolene sutures. PMID- 11420169 TI - New physiological and pathophysiological aspects on the thromboxane A(2) prostacyclin regulatory system. PMID- 11420170 TI - Accurate separation of vesicles, micelles and cholesterol crystals in supersaturated model biles by ultracentrifugation, ultrafiltration and dialysis. AB - Gel filtration with bile salts at intermixed micellar/vesicular concentrations (IMC) in the eluant has been proposed to isolate vesicles and micelles from supersaturated model biles, but the presence of vesicular aggregates makes this method unreliable. We have now validated a new method for isolation of various phases. First, aggregated vesicles and - if present - cholesterol crystals are pelleted by short ultracentrifugation. Cholesterol contained in crystals and vesicular aggregates can be quantitated from the difference of cholesterol contents in the pellets before and after bile salt-induced solubilization of the vesicular aggregates. Micelles are then isolated by ultrafiltration of the supernatant through a highly selective 300 kDa filter and unilamellar vesicles by dialysis against buffer containing bile salts at IMC values. Lipids contained in unilamellar vesicles are also estimated by subtraction of lipid contents in filtered micelles from lipid contents in (unilamellar vesicle+micelle containing) supernatant ('subtraction method'). 'Ultrafiltration-dialysis' and 'subtraction' methods yielded identical lipid solubilization in unilamellar vesicles and identical vesicular cholesterol/phospholipid ratios. In contrast, gel filtration yielded much more lipids in micelles and less in unilamellar vesicles, with much higher vesicular cholesterol/phospholipid ratios. When vesicles obtained by dialysis were analyzed by gel filtration, vesicular cholesterol/phospholipid ratios increased strongly, despite correct IMC values for bile salts in the eluant. Subsequent extraction of column material showed significant amounts of lipids. In conclusion, gel filtration may underestimate vesicular lipids and overestimate vesicular cholesterol/phospholipid ratios, supposedly because of lipids remaining attached to the column. Combined ultracentrifugation ultrafiltration-dialysis should be considered state-of-the-art methodology for quantification of cholesterol carriers in model biles. PMID- 11420171 TI - Isoprenoid biosynthesis is not compromised in a Zellweger syndrome mouse model. AB - Because several studies indicated that peroxisomes are important for the biosynthesis of isoprenoids, we wanted to investigate whether a reduced availability of isoprenoids could be one of the pathogenic factors contributing to the severe phenotype of the Pex5(-/-) mouse, a model for Zellweger syndrome. Total cholesterol was determined in plasma, brain and liver of newborn mice. In none of these tissues a significant difference was observed between Pex5(-/-) and wild type or heterozygous mice. The hepatic ubiquinone content was found to be even higher in Pex5(-/-) mice as compared to wild type or heterozygous littermates. To investigate whether the Pex5(-/-) fetuses are able to synthesise their own isoprenoids, fibroblasts derived from these mice were incubated with radiolabeled mevalonolactone as a substrate for isoprenoid synthesis. No significant difference was observed between the cholesterol production rates of Pex5(-/-) and normal fibroblasts. Our results show that there is no deficiency of isoprenoids in newborn Pex5(-/-) mice, excluding the possibility that a lack of these compounds is a determinant factor in the development of the disease state before birth. PMID- 11420172 TI - Subcellular study of sphingoid base phosphorylation in rat tissues: evidence for multiple sphingosine kinases. AB - The enzymatic phosphorylation of sphingoid bases was analysed in rat tissues, using D-erythro-[4,5-(3)H]sphinganine as substrate. After optimisation of the assay, taking care to block sphingosine-phosphate lyase and sphingosine phosphatase, highest ATP-dependent kinase activities were present in testis, followed by kidney, and intestinal mucosa. Approximately two thirds of the kidney activity were membrane bound, the remaining being cytosolic. Classical cell fractionation studies of kidney and liver did not allow to identify unequivocally the subcellular site of the membrane bound kinase. Separation of a particulate fraction from kidney homogenates by Percoll gradient and sucrose density gradient centrifugation revealed that kinase activities are associated with vesicles derived from the endoplasmic reticulum and the plasma membrane. Based on indirect data, such as the effect of detergents and divalent ions, the cytosolic and both membrane bound activities appear to reside in different proteins. N,N Dimethylsphingenine was inhibitory to all three different kinases, which were mainly active towards the D-erythro isomers of sphingenine and sphinganine. PMID- 11420173 TI - Is a role of phospholipase A(2) in cholesterol gallstone formation phospholipid species-dependent? AB - Phospholipase A(2) plays a role in cholesterol gallstone formation by hydrolyzing bile phospholipids into lysolecithin and free fatty acids. This study investigated its effects on cholesterol crystallization in model bile systems. Supersaturated model bile solutions with different cholesterol saturation indexes (1.2, 1.4, and 1.6) were prepared using cholesterol, taurocholate, and egg yolk phosphatidylcholine, soybean phosphatidylcholine, palmitoyl-oleoyl phosphatidylcholine, or palmitoyl-linoleoyl phosphatidylcholine. Then the effect of digestion of phosphatidylcholine by phospholipase A(2) on bile metastability was assessed by spectrophotometry and video-enhanced differential contrast microscopy. Addition of phospholipase A(2) caused the release of free fatty acids in a time-dependent manner. Cholesterol crystallization was enhanced by an increased crystal growth rate in model bile containing hydrophilic species such as soybean or palmitoyl-linoleoyl phosphatidylcholine, consisting predominantly of polyunsaturated fatty acids. Because phospholipase A(2) enhanced cholesterol crystallization in bile containing hydrophilic phosphatidylcholine species, but not hydrophobic phosphatidylcholine species, release of polyunsaturated fatty acids by hydrolysis may be responsible for such enhancement. Therefore, the role of phospholipase A(2) in cholesterol gallstone formation depends on the phospholipid species present in bile, so that phospholipid species selection during hepatic excretion is, in part, crucial to the cholesterol stone formation. PMID- 11420174 TI - Platelet-activating factor (PAF)-acetylhydrolase and PAF-like compounds in the lung: effects of hyperoxia. AB - Platelet-activating factor (PAF)-acetylhydrolase is the enzyme modulating in tissues and biological fluids the concentration of the proinflammatory factors PAF and PAF-like oxidation products of phospholipids (PAF-like compounds). We investigated whether there is a relation between PAF-acetylhydrolase activity and the concentration of PAF-like compounds in bronchoalveolar lavage (BAL). We found that alveolar type II cells are an additional source of PAF-acetylhydrolase in BAL beside macrophages. Secretion of PAF-acetylhydrolase was stimulated by phorbol ester in alveolar type II cells but not in macrophages. Studies in BAL suggested that secreted PAF-acetylhydrolase was bound to alveolar surfactant. Exposure of rats to high oxygen concentration reduced the activity of PAF acetylhydrolase in BAL and macrophages, but not in plasma or alveolar type II cells. In contrast, hyperoxia increased the concentration of PAF-like-compounds, lipid hydroperoxides and malonedialdehyde in plasma but not in BAL. Therefore, we conclude that neither the oxidant-induced decrease of the PAF-acetylhydrolase activity nor the direct peroxidation of surfactant lipids in the alveoli provide a likely mechanism for hyperoxia-induced lung injury. Instead, lung injury is apparently caused by lipid peroxidation in plasma rather than by high oxygen pressure in the alveoli. PMID- 11420175 TI - Expression, purification, and characterization of the Mycobacterium tuberculosis acyl carrier protein, AcpM. AB - Mycolic acids are generated in Mycobacterium tuberculosis as a result of the interaction of two fatty acid biosynthetic systems: the multifunctional polypeptide, FASI, in which the acyl carrier protein (ACP) domain forms an integral part of the polypeptide, and the dissociated FASII system, which is composed of monofunctional enzymes and a discrete ACP (AcpM). In order to characterize enzymes of the FASII system, large amounts of AcpM are required to generate substrates such as holo-AcpM, malonyl-AcpM and acyl-AcpM. The M. tuberculosis acpM gene was overexpressed in Escherichia coli and AcpM purified, yielding approximately 15-20 mg/l of culture. Analysis of AcpM by mass spectrometry, N-terminal sequencing, amino acid analysis, and gas chromatography indicated the presence of three species, apo-, holo-, and acyl-AcpM, the former comprising up to 65% of the total pool. The apo-AcpM was purified away from the in vivo generated holo- and acyl-forms, which were inseparable and heterogeneous with respect to acyl chain lengths. Once purified, we were able to convert apo AcpM into holo- and acyl-forms. These procedures provide the means for the preparation of the large quantities of AcpM and derivatives needed for characterization of the purified enzymes of the mycobacterial FASII system. PMID- 11420176 TI - Production of acetate in the liver and its utilization in peripheral tissues. AB - In experimental rat liver perfusion we observed net production of free acetate accompanied by accelerated ketogenesis with long-chain fatty acids. Mitochondrial acetyl-CoA hydrolase, responsible for the production of free acetate, was found to be inhibited by the free form of CoA in a competitive manner and activated by reduced nicotinamide adenine dinucleotide (NADH). The conditions under which the ketogenesis was accelerated favored activation of the hydrolase by dropping free CoA and elevating NADH levels. Free acetate was barely metabolized in the liver because of low affinity, high K(m), of acetyl coenzyme A (acetyl-CoA) synthetase for acetate. Therefore, infused ethanol was oxidized only to acetate, which was entirely excreted into the perfusate. The acetyl-CoA synthetase in the heart mitochondria was much lower in K(m) than it was in the liver, thus the heart mitochondria was capable of oxidizing free acetate as fast as other respiratory substrates, such as succinate. These results indicate that rat liver produces free acetate as a byproduct of ketogenesis and may supply free acetate, as in the case of ketone bodies, to extrahepatic tissues as fuel. PMID- 11420177 TI - A role for hormone-sensitive lipase in the selective mobilization of adipose tissue fatty acids. AB - The mobilization of fatty acids from rat and human fat cells is selective according to molecular structure, and notably carbon atom chain length. This study aimed at examining whether the release of individual fatty acids from triacylglycerols (TAG) by hormone-sensitive lipase (HSL) plays a role in the selectivity of fatty acid mobilization. Recombinant rat and human HSL were incubated with a lipid emulsion. The hydrolysis of 18 individual fatty acids, ranging in chain length from 12 to 24 carbon atoms and in unsaturation degree from 0 to 3 double bond(s), was measured by comparing the composition of non esterified fatty acids (NEFA) to that of the original TAG. The relative hydrolysis (% in NEFA/% in TAG) differed between fatty acids, being about 5-fold and 3-fold higher for the most (18:1n-7) than for the least (24:0) readily released fatty acid by recombinant rat and human HSL, respectively. Relationships were found between the chain length of fatty acids and their relative hydrolysis. Among 12-24 carbon atom saturated fatty acids, the relative hydrolysis markedly decreased (by about 5- and 3-times for recombinant rat and human HSL, respectively) with increasing chain length. We conclude that fatty acids are selectively released from TAG by HSL according to carbon atom chain length. These data provide insight on the mechanism by which fatty acids are selectively mobilized from fat cells. PMID- 11420178 TI - Adipophilin is a sensitive marker for lipid loading in human blood monocytes. AB - Adipophilin, a marker of lipid accumulation initially described in adipocytes, was recently shown to be induced in macrophage foam cells. We found that even freshly isolated blood monocytes express adipophilin and that the amount of adipophilin protein is variable in monocytes from different healthy individuals. However, the physiological expression of adipophilin does not correlate with the levels of free fatty acids, cholesterylesters or free cholesterol. Enzymatically modified low-density lipoprotein (E-LDL) induces rapid foam cell formation in monocytes and upregulates adipophilin mRNA and protein within 2 h of incubation. This rapid induction of adipophilin is accompanied by a significant increase of free fatty acids in monocytes incubated with E-LDL. Adipophilin facilitates the uptake of free fatty acids, and here we demonstrate that free fatty acids increase is related to the early upregulation of adipophilin expression in blood monocytes. Fatty acids are ligands for peroxisome proliferator-activated receptor gamma (PPARgamma), and the upregulation of adipophilin mRNA by PPARgamma agonists like 15d-PGJ(2) and ciglitazone indicates that PPARgamma may mediate the induction of adipophilin expression in human blood monocytes. PMID- 11420179 TI - Structure, expression profile and alternative processing of the human phosphatidylethanolamine N-methyltransferase (PEMT) gene. AB - Phosphatidylethanolamine N-methyltransferase (PEMT) catalyzes the conversion of phosphatidylethanolamine (PE) to phosphatidylcholine (PC) in a series of three methylation reactions. Preliminary studies of PEMT in humans led to the cloning of three cDNAs each of which has a different 5' untranslated region (5'UTR). To determine the origin of PEMT splice variants and to investigate expression of the gene in human liver, we isolated a bacterial artificial chromosome (BAC) clone containing the full-length human gene. Each of the three unique untranslated first exons is present in a contiguous array in the gene, confirming the integrity of the cDNAs and alternative processing of PEMT transcripts. Human liver, heart and testis contain the highest levels of PEMT transcripts and of these, liver has the greatest PEMT expression. Furthermore, each of the three PEMT transcripts is present in varying abundance in liver whereas heart and testis contain only one and two transcripts, respectively. Thus, differential promoter usage in the human PEMT gene generates three unique transcripts and confers a tissue-specific expression pattern. PMID- 11420180 TI - Increase of ceramides and its inhibition by catalase during chemically induced apoptosis of HL-60 cells determined by electrospray ionization tandem mass spectrometry. AB - A change in all ceramide species during chemically induced apoptosis of HL-60 cells was determined using electrospray tandem mass spectrometry. Ceramides of C16:0, C24:1 and C26:1 increased significantly 4 h after the addition of actinomycin D, when the activation of caspase-3 was maximal. Addition of catalase, which inhibited apoptosis, the activation of caspase-3-like protease, and the release of cytochrome c from mitochondria to cytosol caused by actinomycin D or daunorubicin, significantly inhibited the increase of these ceramides at all time points. Ceramides of C16:0, C24:1, C18:0, C22:1 and C26:1 increased significantly 4 h after the addition of daunorubicin to HL-60 cells. Catalase also significantly inhibited the increase of these ceramides induced by daunorubicin. Based on time courses of events and inhibition studies, it is concluded that the increase of ceramides is downstream from both generation of hydrogen peroxide and cytochrome c release from mitochondria and takes place almost simultaneously with the activation of caspase-3. PMID- 11420181 TI - Marked activation of the N-acylphosphatidylethanolamine-hydrolyzing phosphodiesterase by divalent cations. AB - N-Acylethanolamines including anandamide (an endogenous ligand for cannabinoid receptors) are released from N-acylphosphatidylethanolamine (N-acyl-PE) by the catalysis of a phosphodiesterase of the phospholipase D type. The enzyme was solubilized from the particulate fractions of rat heart with the aid of octyl glucoside, and partially purified by anion-exchange chromatography. The enzyme hydrolyzed N-palmitoyl-PE with a specific activity of 17 nmol/min/mg protein at 37 degrees C. The enzyme activity increased dramatically up to 30-fold by millimolar order of Ca(2+). Ca(2+) could be replaced with other divalent cations such as Co(2+), Mg(2+), Mn(2+), Ba(2+), Sr(2+) and Ni(2+). The hydrolysis of N arachidonoyl-PE (a precursor of anandamide) was also markedly stimulated by Ca(2+). PMID- 11420182 TI - Interplay in lipoplexes between type of pDNA promoter and lipid composition determines transfection efficiency of human growth hormone in NIH3T3 cells in culture. AB - This study was aimed to investigate if and to what extent there is an interplay between lipoplex physicochemical properties and plasmid promoter type affecting transfection efficiency in vitro. To reduce the number of variables only one cell type (NIH3T3 cells), one gene (human growth hormone), one cationic lipid (DOTAP) in a plasmid >85% in supercoiled form, and the same medium conditions were used. The variables of the physicochemical properties included presence and type of helper lipid (DOPE, DOPC, or cholesterol, all in 1:1 mole ratio with DOTAP), size and lamellarity of the liposomes used for lipoplex preparation (large unilamellar vesicles, LUV, versus multilamellar vesicles, MLV), and DNA(-)/cationic lipid(+) charge ratio, all containing the same human growth hormone but differing in their promoter enhancer region. Two of the promoters were of viral origin: (a) SV40 promoter (simian virus early promoter) and (b) CMV promoter (cytomegalovirus early promoter); two were of mammalian cell origin: (c) PABP promoter (human poly(A)-binding protein promoter) and (d) S16 promoter (mouse ribosomal protein (rp) S16 promoter). Transfection studies showed that, irrespective of promoter type, large (> or =500 nm) MLV were superior to approximately 100 nm LUV; the extent of superiority was dependent on liposome lipid composition (larger for 100% DOTAP and DOTAP/DOPE than for DOTAP/DOPC and DOTAP/cholesterol). The optimal DNA(-)/DOTAP(+) charge ratio for all types of lipoplexes used was 0.2 or 0.5 (namely, when the lipoplexes were positively charged). Scoring the six best lipoplex formulations (out of 128 studied) revealed the following order: pCMV (DOTAP/DOPE) >> pSV (DOTAP/DOPE)=pCMV(DOTAP/cholesterol)=pS16 (100% DOTAP)=pS16 DOTAP/DOPE >> pCMV (DOTAP/DOPC). The lack of trivial consistency in the transfection efficiency score, the pattern of transfection efficiency, and statistical analysis of the data suggest that there is cross-talk between promoter type and lipoplex lipid composition, which may be related to the way the promoter is associated with the lipids. PMID- 11420183 TI - Differences in the conversion of the polyunsaturated fatty acids [1-(14)C]22:4(n 6) and [1-(14)C]22:5(n-3) to [(14)C]22:5(n-6) and [(14)C]22:6(n-3) in isolated rat hepatocytes. AB - The reasons why most cellular lipids preferentially accumulate 22:6(n-3) rather than 22:5(n-6) are poorly understood. In the present work the metabolisms of the precursor fatty acids, [1-(14)C]20:4(n-6), [1-(14)C]22:4(n-6) versus [1 (14)C]20:5(n-3), [1-(14)C]22:5(n-3) in isolated rat hepatocytes were compared. The addition of lactate and L-decanoylcarnitine increased the formation of [(14)C]24 fatty acid intermediates and the final products, [(14)C]22:5(n-6) and [(14)C]22:6(n-3). In the absence of lactate and L-decanoylcarnitine, no [(14)C]24 fatty acids and [(14)C]22:5(n-6) were detected when [1-(14)C]22:4(n-6) was the substrate, whereas small amounts of the added [1-(14)C]22:5(n-3) was converted to [(14)C]22:6(n-3). Lactate reduced the oxidation of [1-(14)C]22:4(n-6) and [1 (14)C]22:5(n-3) while L-decanoylcarnitine did not. No significant differences between the total oxidation or esterification of the two substrates were observed. By fasting and fructose refeeding the amounts of [(14)C]24:4(n-6) and [(14)C]24:5(n-3) were increased by 2.5- and 4-fold, respectively. However, the levels of [(14)C]22:5(n-6) and [(14)C]22:6(n-3) were similar in hepatocytes from fasted and refed versus fed rats. With hepatocytes from rats fed a fat free diet the levels of [(14)C]24 fatty acid intermediates were low while the further conversion of the n-6 and n-3 substrates was high and more equal, approx. 33% of [1-(14)C]22:4(n-6) was converted to [(14)C]22:5(n-6) and 43% of [1-(14)C]22:5(n 3) was converted to [(14)C]22:6(n-3). The moderate differences found in the conversion of [1-(14)C]22:4(n-6) versus [1-(14)C]22:5(n-3) to [(14)C]22:5(n-6) and [(14)C]22:6(n-3), respectively, and the equal rates of oxidation of the two substrates could thus not explain the abundance of 22:6(n-3) versus the near absence of 22:5(n-6) in cellular membranes. PMID- 11420184 TI - Getting ADC to paediatricians in developing countries. PMID- 11420185 TI - Alpha-1 antitrypsin deficiency. PMID- 11420186 TI - Neurodevelopmental outcome in meningococcal disease: a case-control study. AB - AIMS: To determine long term neurodevelopmental outcome following the spectrum of meningococcal infection. METHODS: Between 1988 and 1990, 152 cases of meningococcal disease were recruited; 139 survived. Between 1998 and 1999, 115 survivors (83%) were evaluated, together with 115 sex and age matched controls. Standard measures of neurological function, coordination, cognition, behaviour, and hearing were used to assess neurodevelopmental status. RESULTS: One case has spastic quadriplegia. Gross neurological examination was normal in all other cases and all controls. Five cases and no controls have significant hearing loss. Cases performed at a lower level than controls on measures of coordination, cognition, and behaviour. Four cases and no controls had major impairments. The adjusted odds ratios for moderate and minor impairments were 3.6 (95% CI 1.3 to 10.3) and 1.6 (95% CI 0.8 to 3.4) respectively. CONCLUSION: The majority of survivors from this cohort do not have gross neurological deficits. However, when objective measures of motor function, cognitive ability, and behaviour were applied significant detriments were found in meningococcal survivors. PMID- 11420187 TI - What's up (or down and out) in US healthcare. PMID- 11420188 TI - Invasive procedures carried out in conscious children: contrast between North American and European paediatric oncology centres. AB - AIM: To define practice in managing repeated invasive procedures in selected paediatric oncology centres in North America and Europe, especially the United Kingdom; to define and contrast concerns that shape policy making, and to contrast practice, particularly regarding procedures performed on conscious patients. METHODS: Postal survey: 118 centres of the Pediatric Oncology Group and the United Kingdom Children's Cancer Study Group received questionnaires. RESULTS: 68 questionnaires (58%) were returned (52 from North America, 12 from Europe). For all procedures, North American centres tended to use less effective techniques than European, especially for bone marrow procedures. Many North American centres reported performing these on conscious patients on at least three quarters (25%) or half (30%) the occasions. In contrast, corresponding figures for the European centres were 6% and 0%. CONCLUSIONS: Many bone marrow procedures are still carried out in the conscious patient despite the safety and effectiveness of modern anaesthetic and deep sedation techniques. There appears to be a greater reluctance to offer these to patients in North American centres than in European ones. This may reflect a misperception that the risks of adverse effects are high. Several non-pharmacological techniques are used, but they remain uncommon. PMID- 11420189 TI - The risk and outcome of cerebral oedema developing during diabetic ketoacidosis. AB - BACKGROUND: Cerebral oedema is a major cause of morbidity and mortality in children with insulin dependent diabetes. AIMS: To determine the risk and outcome of cerebral oedema complicating diabetic ketoacidosis (DKA). METHODS: All cases of cerebral oedema in England, Scotland, and Wales were reported through the British Paediatric Surveillance Unit between October 1995 and September 1998. All episodes of DKA were reported by 225 paediatricians identified as involved in the care of children with diabetes through a separate reporting system between March 1996 and February 1998. Further information about presentation, management, and outcome was requested about the cases of cerebral oedema. The risk of cerebral oedema was investigated in relation to age, sex, seasonality, and whether diabetes was newly or previously diagnosed. RESULTS: A total of 34 cases of cerebral oedema and 2940 episodes of DKA were identified. The calculated risk of developing cerebral oedema was 6.8 per 1000 episodes of DKA. This was higher in new (11.9 per 1000 episodes) as opposed to established (3.8 per 1000) diabetes. There was no sex or age difference. Cerebral oedema was associated with a significant mortality (24%) and morbidity (35% of survivors). CONCLUSIONS: This first large population based study of cerebral oedema complicating DKA has produced risk estimates which are more reliable and less susceptible to bias than those from previous studies. Our study indicates that cerebral oedema remains an important complication of DKA during childhood and is associated with significant morbidity and mortality. Little is known of the aetiology of cerebral oedema in this condition and we are currently undertaking a case control study to address this issue. PMID- 11420191 TI - Total serum IgE and outcome in infants with recurrent wheezing. AB - OBJECTIVE: To investigate the relation between total serum IgE at 0.5-3 and 3-6 years, and the risk of allergic sensitisation and persistent wheezing up to 8 years of age. METHODS: Prospective follow up study of 45 infants with highly recurrent wheezing, no allergic symptoms, and negative skin tests. RESULTS: In the last follow up year, 15 children still suffered from wheezing. Five wheeze free and four episodically wheezing children had become sensitised. No association was found between early (0.5-3 years) IgE z scores and the recurrence of wheezing during follow up, or atopic sensitisation. IgE z scores at 3-6 years were significantly higher in children with positive skin tests (p = 0.013), but were still not associated with recurrence of wheezing. CONCLUSIONS: In subjects with frequent early wheezing and no signs of atopy, early total serum IgE measurements are not predictive of outcome. PMID- 11420193 TI - Congenital adrenal hyperplasia: management during critical illness. AB - BACKGROUND: Little is known of the optimal dose and administration schedule of hydrocortisone in critically ill patients with congenital adrenal hyperplasia (CAH) caused by 21-hydroxylase deficiency. AIM: To determine plasma cortisol concentrations after intravenous administration of hydrocortisone in children with CAH and to relate these to plasma cortisol concentrations achieved by endogenous secretion in the stress of critical illness in previously healthy children. METHODS: Plasma cortisol concentrations were measured in 20 patients with classical CAH (median age 11.2 years, range 6.1-16.4) following intravenous administration of hydrocortisone 15 mg/m(2); and in 60 critically ill mechanically ventilated children (median age 2.5 years, range 0.25-16.3) on admission to the paediatric intensive care unit and for 24 hours thereafter. RESULTS: In the CAH patients, plasma cortisol reached a mean peak of 1648.3 nmol/l (SD 511.9) within 10 minutes of the intravenous bolus, and fell rapidly thereafter; levels remained greater than 450 nmol/l for 2.5 hours only. In critically ill children, mean plasma cortisol on admission to the intensive care unit was 727 nmol/l (SD 426.1). Cortisol concentrations remained raised during the first 24 hours. CONCLUSIONS: Critically ill patients with classical CAH may be best managed with a single intravenous hydrocortisone bolus followed by a constant rate infusion of hydrocortisone. PMID- 11420195 TI - Characterisation of breathing and associated central autonomic dysfunction in the Rett disorder. AB - AIM: To investigate breathing rhythm and brain stem autonomic control in patients with Rett disorder. SETTING: Two university teaching hospitals in the United Kingdom and the Rett Centre, Sweden. PATIENTS: 56 female patients with Rett disorder, aged 2-35 years; 11 controls aged 5-28 years. DESIGN: One hour recordings of breathing movement, blood pressure, ECG R-R interval, heart rate, transcutaneous blood gases, cardiac vagal tone, and cardiac sensitivity to baroreflex measured on-line with synchronous EEG and video. Breathing rhythms were analysed in 47 cases. RESULTS: Respiratory rhythm was normal during sleep and abnormal in the waking state. Forced and apneustic breathing were prominent among 5-10 year olds, and Valsalva breathing in the over 18 year olds, who were also most likely to breathe normally. Inadequate breathing peaked among 10-18 year olds. Inadequate and exaggerated breathing was associated with vacant spells. Resting cardiac vagal tone and cardiac sensitivity to baroreflex were reduced. CONCLUSIONS: Labile respiratory rhythms and poor integrative inhibition in Rett disorder suggest brain immaturity. Linking this to an early monoaminergic defect suggests possible targets for the MECP2 gene in clinical intervention. Exaggerated and inadequate autonomic responses may contribute to sudden death. PMID- 11420196 TI - Bile bilirubin pigment analysis in disorders of bilirubin metabolism in early infancy. AB - BACKGROUND: Early and accurate diagnosis of Crigler-Najjar syndrome, which causes prolonged unconjugated hyperbilirubinaemia in infancy, is important, as orthotopic liver transplantation is the definitive treatment. AIM: To determine whether bilirubin pigment analysis of bile in infants with prolonged unconjugated hyperbilirubinaemia provides useful diagnostic information in the first 3 months of life. METHODS: Retrospective review of patients with prolonged unconjugated hyperbilirubinaemia referred to the liver unit, Birmingham Children's Hospital, for the diagnosis of Crigler-Najjar syndrome. Bile bilirubin pigment composition was determined by high performance liquid chromatography. Initial diagnoses were made based on the result of bile bilirubin pigment composition. Final diagnoses were made after reviewing the clinical course, response to phenobarbitone, repeat bile bilirubin pigment composition analysis, and genetic studies. RESULTS: Between 1992 and 1999, nine infants aged less than 3 months of age with prolonged hyperbilirubinaemia underwent bile bilirubin pigment analyses. Based on these, two children were diagnosed with Crigler-Najjar syndrome (CNS) type 1, six with CNS type 2, and one with Gilbert's syndrome. Five children whose initial diagnosis was CNS type 2 had resolution of jaundice and normalisation of serum bilirubin after discontinuing phenobarbitone, and these cases were thought to be normal or to have Gilbert's syndrome. One of the initial cases of CNS type 1 responded to phenobarbitone with an 80% reduction in serum bilirubin consistent with CNS type 2. In all, the diagnoses of six cases needed to be reviewed. CONCLUSIONS: Early bile pigment analysis, performed during the first 3 months of life, often shows high levels of unconjugated bilirubin or bilirubin monoconjugates, leading to the incorrect diagnosis of both type 1 and type 2 Crigler-Najjar syndrome. PMID- 11420197 TI - 99mTc-HMPAO leucocyte scintigraphy fails to detect Crohn's disease in the proximal gastrointestinal tract. AB - OBJECTIVE: To investigate the use of (99m)Tc-HMPAO (hexamethyl propylene amine oxime) leucocyte scintigraphy as a non-invasive screening test for inflammatory bowel disease. PATIENTS: 10 children with suspected Crohn's disease, in whom routine investigation using barium contrast radiology, upper gastrointestinal endoscopy, colonoscopy, and mucosal biopsies had identified severe gastroduodenal and/or jejunal involvement. DESIGN: (99m)Tc-HMPAO leucocyte scintigraphic studies performed in each of these cases were assessed by a radiologist who was blinded to the disease distribution. RESULTS: In nine cases there was no scintigraphic evidence of inflammation in the proximal gastrointestinal tract. The 10th child had both gastroduodenal and jejunal involvement, but scintigraphy only revealed faint jejunal positivity. CONCLUSIONS: (99m)Tc-HMPAO leucocyte scintigraphy should not be depended upon as a screening test for Crohn's disease. False negative results are likely in cases with Crohn's disease confined to the proximal gastrointestinal tract. PMID- 11420198 TI - World wide wasting? PMID- 11420199 TI - Hyperhomocysteinaemia and MTHFR C677T gene polymorphism in renal transplant recipients. AB - AIM: To study the effect of folate treatment on hyperhomocysteinaemia and the effect of 5,10-methylenetetrahydrofolate reductase (MTHFR) gene polymorphism on total homocysteine and folate concentrations after renal transplantation. METHODS: A total of 30 transplanted children and adolescents were investigated for total homocysteine and folate serum concentrations before and after folate treatment, as well as for the presence of the MTHFR C677T polymorphism. RESULTS: The allele frequency of C677T polymorphism in the MTHFR gene in the study population (0.33) was not different to that in controls (0.38). Before folate treatment the homocysteine concentration was raised in all groups; following folate supplementation it was significantly decreased in the CC and CT groups, but not in the TT group. In patients with CC genotype, serum homocysteine correlated with serum creatinine and cholesterol, and time since transplantation before treatment. CONCLUSION: Folate supplementation appears to be an effective strategy to normalise total homocysteine concentration in renal transplanted children and adolescents. PMID- 11420200 TI - Impact of HIV-1 infection in South Africa. AB - In a prospective study, 60% of admissions to an academic hospital were infected with HIV. HIV infected children were younger, less likely to have been referred, more likely to have pneumonia and candidiasis, and had more health service attendances. This impact may be alleviated by appropriate primary and secondary level health care. PMID- 11420201 TI - Management of sickle cell priapism with etilefrine. AB - Intracavernous injections of etilefrine were effective in seven children with acute sickle cell priapism, and stuttering priapism resolved in five children after one to seven months of oral etilefrine. Compared with our previous reports in adults, etilefrine appears to be more effective in childhood. PMID- 11420202 TI - Facial nerve palsy associated with Rickettsia conorii infection. AB - Facial nerve palsy has been occasionally attributed to infectious agents, but Rickettsiae species have not been documented as causative agents. We report two adolescent girls with facial nerve palsy and serological evidence of R conorii infection. These cases indicate that rickettsioses should be included among the causes of facial nerve palsy, particularly in endemic areas. PMID- 11420203 TI - Intussusception associated with bacterial meningitis. AB - Despite its common association with viral illnesses, intussusception has only rarely been found in the presence of bacterial infections. Two infants are described, both of whom were admitted to hospital with bilious vomiting, drowsiness, and dehydration. Both infants required urgent intravenous volume expansion. Intussusception was confirmed, and reduction was achieved by enema in both cases. Recovery was slow, and one infant developed a seizure. Evidence of meningococcal meningitis was found in both, with septicaemia in one. Neurological outcome is normal to date, and there has been no recurrence of intussusception in either case. PMID- 11420204 TI - Urinary calcium excretion in enterally fed disabled children. AB - The urine calcium/creatinine (Ca/Cr) ratio was measured in 17 enterally tube fed disabled children. Urine Ca/Cr ratios were inversely related to dietary calcium intake in immobile children (r = -0.57, p < 0.05) but not in those who were able to stand or walk (mobile children) (r = 0.4, p = 0.51). None of the subjects had evidence of nephrocalcinosis or renal calculi detectable by renal ultrasonography. PMID- 11420205 TI - Diabetic ketoacidosis in Asian children. AB - Over the 10 year period 1987-1996, 328 children with type 1 diabetes mellitus presented in the city of Birmingham, England, of whom 27% had diabetic ketoacidosis. Asian children under the age of 5 had an eightfold increased risk of presenting in diabetic ketoacidosis compared with non-Asian children of the same age. PMID- 11420207 TI - Recent advances in cystic fibrosis. AB - The median life expectancy for cystic fibrosis is now over 30 years, and it is projected that in newborn infants it will become more than 40 years. The identification of the cystic fibrosis gene and its product, cystic fibrosis transmembrane conductance regulator (CFTR), has widened the spectrum of the disease from the classical case of the infant with cystic fibrosis to the elderly childless man with unexplained bronchiectasis. There is increasing evidence of the advantages of newborn screening for cystic fibrosis and subsequent specialist care. Management concentrates on optimising nutritional status and preventing lung infection and inflammation. PMID- 11420208 TI - A critique of the expression of paediatric body composition data. AB - There is increasing interest in body composition in paediatric research, as distinct from growth and nutritional status, as almost all diseases have adverse effects on either fatness or the fat-free mass. However, the approaches used to assess growth and nutritional status are not appropriate for separate evaluations of body fatness and lean mass. Traditional measurements such as body mass index and skinfold thickness do not measure fat in accurate quantitative terms. Various techniques have been used in recent years which divide body weight into fat mass and fat-free mass; however, the data tend not to be appropriately expressed. Body fatness is generally expressed as a percentage of weight, while fat-free mass typically remains unadjusted for size. A more appropriate approach is to normalise both body fatness and fat-free mass for height. This recommendation is relevant both to studies comparing patients with controls and to the expression of new reference data on body composition which are needed to allow informative comparisons. The same approach is appropriate for the classification of childhood obesity. PMID- 11420210 TI - Energy metabolism in relation to body composition and gender in adolescents. AB - OBJECTIVE: To study the effect of body composition on average daily metabolic rate (ADMR) and basal metabolic rate (BMR) in adolescence, and to examine current BMR prediction equations. STUDY: Dutch adolescents were pooled with previously reported American and British subjects (n = 90 overall). BMR and ADMR were analysed by multiple regression. RESULTS: Fat-free mass, BMR, and ADMR were higher in the obese than in the non-obese group (mean (SD): 53.2 (10.7) kg, 8.35 (1.57) MJ/d, and 13.64 (2.78) MJ/d, compared with 41.0 (8.1) kg, 6.42 (0.94) MJ/d, and 11.16 (2.21) MJ/d, respectively). BMR remained higher when adjusted for fat-free mass, age, and sex. ADMR adjusted for BMR was similar in the two groups. WHO equations overestimated BMR in obese boys and underestimated BMR in non-obese boys. CONCLUSIONS: BMR, but not activity, is increased in obese adolescents and in male adolescents. The WHO BMR equations for adults are recommended for obese adolescents. PMID- 11420216 TI - Expression profiles of endometrial leukemia inhibitory factor, transforming growth factor beta2 (TGFbeta2), and TGFbeta2 receptor in infertile bonnet monkeys. AB - The expression profiles of leukemia inhibitory factor (LIF), transforming growth factor beta2 (TGFbeta2), and transforming growth factor beta2 receptor (TGFbeta2R) were analyzed during the peri-implantation period in regularly menstruating, fertile bonnet monkeys and in animals in which endometrial nonreceptivity was induced by administering an antiprogestin, onapristone. Based on our previous experiences, a dose of 2.5 or 5 mg of onapristone was administered s.c. every third day during the menstrual cycle, because these dosages impair endometrial development without upsetting the normal gonadal endocrine profiles. Endometrial biopsy specimens were collected during the proliferative phase (estradiol levels about 200 pg/ml, n = 5) and peri implantation period (Day 8 after midcycle peak in estradiol levels, n = 5) from normal ovulatory animals and during the peri-implantation period from onapristone treated animals (n = 10). The biopsy specimens were processed to determine the expression patterns of LIF, TGFbeta2, and TGFbeta2R by immunohistochemical and reverse transcription-polymerase chain reaction (RT-PCR) methods. Levels of both protein and mRNA for LIF, TGFbeta2, and TGFbeta2R (analyzed by immunohistochemistry and RT-PCR, respectively) were greater in the endometrial samples collected during the peri-implantation period compared to samples collected during the proliferative phase in control animals. Treatment with either of the two doses (2.5 or 5 mg) of onapristone caused a significant (P < 0.05) down-regulation in the expression of LIF in the peri-implantation endometria. The endometrial expressions of TGFbeta2 and TGFbeta2R mRNAs were reduced significantly in animals treated with 5 mg of onapristone, but not in those treated with the lower dose. However, immunoreactive TGFbeta2 and TGFbeta2R proteins were significantly (P < 0.05) down-regulated in the endometrial samples from both the 2.5- and 5-mg-treated groups. The alterations observed in the expression patterns of LIF, TGFbeta2, and TGFbeta2R were specific, because the expression levels of epidermal growth factor receptor remained unaffected in the endometria from the treated groups. The present study demonstrates derangement in the expression profiles of LIF, TGFbeta2, and TGFbeta2R during the peri implantation period in infertile bonnet monkeys. It may be hypothesized that TGFbeta2 function is one of the early steps in the regulation of the progesterone driven cascade of events leading to endometrial receptivity, and that any aberration in this step may adversely affect the subsequent molecular events (i.e., expression of LIF). These data also suggest that potential aberrations in the functional network of locally produced cytokines and growth factors even may occur in an endometrium exposed to the optimal peripheral hormonal levels. PMID- 11420217 TI - Efficient in vitro production of cat embryos in modified synthetic oviduct fluid medium: effects of season and ovarian status. AB - The effects of season (January-March = I; April-June = II; July-September = III; October-December = IV) and ovarian status (freshly ovulated, follicular, luteal, intermediate, or inactive) on the efficiency of the in vitro production of domestic cat embryos were evaluated. Ovaries and testes from cats with access to daylight were collected at local veterinary clinics. A total of 6843 cumulus oocyte complexes (COCs) were recovered from 363 pairs of ovaries, matured in TCM 199 supplemented with BSA and gonadotropins (IVM), fertilized with epididymal sperm in a medium based on Tyrode albumin lactate pyruvate (IVF), and cultured in synthetic oviduct fluid (SOF) medium supplemented with 10% estrous cow serum (ECS) and essential and nonessential amino acids. The proportion of freshly ovulated, follicular, or luteal ovaries was higher (P < 0.05) in seasons II (64.4%) and III (60.5%) than in seasons I (42.0%) and IV (30.6%). The average number of COCs recovered per donor was not influenced by season. After IVM/IVF, cleavage rates (Day 2) were significantly higher (P < 0.05) in seasons II (mean +/- SEM: 53.1% +/- 1.9%) and III (54.6% +/- 2.8%) than in seasons I (48.4% +/- 1.4%) and IV (44.9% +/- 3.0%). Blastocyst rates on Day 6 were similar in seasons I (25.3% +/- 1.3%), II (28.2% +/- 1.5%), and III (29.6% +/- 2.3%) but were significantly lower (P < 0.01) in season IV (18.6% +/- 2.4%). The corresponding blastocyst rates on Day 8 were 28.9% +/- 1.3%, 33.7% +/- 1.6%, 37.9% +/- 2.3%, and 23.6% +/- 2.6%. In addition, we found a significant effect (P < 0.05) of ovary type; luteal, follicular, and intermediate ovaries yielding a higher proportion of developmentally competent oocytes than did freshly ovulated and inactive ovaries. These data show that the culture system used in our study supports development of IVM/IVF cat oocytes to blastocysts at a higher rate than those obtained with other methods. Although embryos could be produced throughout the year, the efficiency was significantly affected by season and ovary type. PMID- 11420218 TI - Differences in intracellular pH regulation by Na(+)/H(+) antiporter among two cell mouse embryos derived from females of different strains. AB - Regulation of intracellular pH (pH(i)) by two-cell-stage embryos derived from female mice of three different strains (CF-1, Balb/c, and BDF) was investigated. Embryos recovered at a slow rate from intracellular acidosis produced by a pulse of NH(4)Cl; the rate did not differ significantly among strains. Recovery was reversibly inhibited by amiloride or the absence of Na(+), implicating Na(+)/H(+) antiporter activity. The threshold pH(i) (setpoint) below which Na(+)/H(+) antiporter activity was elicited was approximately 7.15 for each strain. No recovery from induced acidosis occurred in the absence of external Na(+) in any strain, and thus embryos could be maintained in acidosis for an extended period. Upon reintroduction of Na(+), embryos derived from either CF-1 or BDF females recovered at a slow rate comparable to that measured in embryos not maintained for a period in Na(+)-free medium, but embryos derived from Balb/c females consistently recovered at a highly accelerated rate. This accelerated recovery appeared to be due, in part, to an activation of the Na(+)/H(+) antiporter in Balb/c-derived embryos, which did not occur in CF-1- or BDF-derived embryos. Thus, embryos derived from different strains of female mice differ in their control of mechanisms for pH(i) regulation. PMID- 11420219 TI - Evaluation of gestational deficiencies in cloned sheep fetuses and placentae. AB - Sheep fetal development at 35 days of gestation was examined following natural mating, in vitro production (IVP) of fertilized embryos, or somatic cell nuclear transfer (NT). Five crossbred (Blackface x Black Welsh) and four purebred (Black Welsh) fetuses and their associated placentae produced by natural mating were morphologically normal and consistent with each other. From 10 ewes receiving 21 IVP embryos, 17 fetuses (81%) were recovered, and 15 of these (88%) were normal. The NT fetuses were derived from two Black Welsh fetal fibroblast cell lines (BLW1 and 6). Transfer of 21 BLW1 and 22 BLW6 NT embryos into 12 and 11 ewes, respectively, yielded 7 (33%) and 8 (36%) fetuses, respectively. Only three (43%) BLW1 and two (25%) BLW6 NT fetuses were normal, with the rest being developmentally retarded. The NT fetal and placental deficiencies included liver enlargement, dermal hemorrhaging, and lack of placental vascular development reflected by reduced or absent cotyledonary structures. Fibroblasts isolated from normal and abnormal cloned fetuses did not differ in their karyotype from sexually conceived fetuses or nuclear donor cell lines. Our results demonstrate that within the first quarter of gestation, cloned fetuses are characterized by a high incidence of developmental retardation and placental insufficiency. These deficiencies are not linked to gross defects in chromosome number. PMID- 11420220 TI - Expression of matrix metalloproteinase-2, -9, -14, and tissue inhibitors of metalloproteinase-1, -2, -3 in the endometrium and placenta of rhesus monkey (Macaca mulatta) during early pregnancy. AB - The extracellular matrix proteolytic machinery has long been recognized as one of the most important mechanisms for regulating trophoblast invasion. Matrix metalloproteinases (MMPs) are a group of proteases involved in this process, and their activities are regulated by tissue inhibitors of MMPs (TIMPs). In this study, we collected rhesus monkey uteri on Days 12, 18, and 26 of pregnancy and examined the mRNA expression of MMP-2, -9, -14, and TIMP-1, -2, -3, as well as the activities of MMP-2 and -9 by using in situ hybridization and gelatin zymography, respectively. The results showed that MMP-2 and -9 were expressed earlier than MMP-14 and TIMPs in the pregnant endometrium. MMP-14 and TIMP-2 mRNAs appeared in perivascular decidual cells earlier than MMP-2 mRNA. On Day 26 of pregnancy, placental villi expressed little MMP-2, -14, and TIMP transcripts but abundant MMP-9 mRNA. Furthermore, MMP-2, -9, -14, and TIMP-1, -2, -3 were highly expressed on the fetal-maternal border but were absent in the myometrium. TIMP-3 mRNA in the endometrium was specifically localized to some cells lining the outer membrane of several groups of arterioles. Combined with the results obtained by gelatin zymography, we found that active MMP-2 existed in the endometrium throughout these three phases, while MMP-9 showed considerable activities only on Days 18 and 26 of pregnancy. The data suggest key roles for MMP-2 and -9 in invasion of trophoblast cells into the endometrium and the development of the placenta and might indicate that these processes are regulated by MMP-14 and TIMP-1, -2, and -3. PMID- 11420221 TI - Effect of seminal phospholipid-binding proteins and follicular fluid on bovine sperm capacitation. AB - Bovine seminal plasma (BSP) contains a family of novel phospholipid-binding proteins (BSP-A1/-A2, BSP-A3, and BSP-30-kDa; collectively called BSP proteins) that potentiate sperm capacitation induced by heparin or by serum high-density lipoprotein (HDL). BSP proteins stimulate lipid efflux from sperm that may occur during the early events of capacitation. Here, we investigated the role of BSP proteins, bovine follicular fluid (FF), and bovine follicular fluid HDL (FF-HDL) in sperm capacitation. FF and FF-HDL alone stimulated epididymal sperm capacitation (19.5% +/- 0.8% and 18.2% +/- 2.8%, respectively, control, 9.0% +/- 1.9%) that was increased by preincubation with BSP-A1/-A2 proteins (30.2% +/- 0.4% and 30.9% +/- 1.5%, respectively). In contrast, lipoprotein-depleted follicular fluid (LD-FF) alone was ineffective, and a preincubation with BSP-A1/ A2 proteins was necessary before sperm capacitation was stimulated (up to 22.8% +/- 1.4%). The interaction of BSP proteins with FF components was analyzed using ultracentrifugation, Lipo-Gel electrophoresis, SDS-PAGE, and gel filtration. We established that the BSP proteins interact with factors present in FF including FF-HDL. Additionally, we obtained evidence that BSP proteins, found associated with FF-HDL, were released from the sperm membrane during capacitation. These results confirm that the BSP proteins and the FF-HDL play a role in sperm capacitation. PMID- 11420222 TI - Ratio of proliferating cell nuclear antigen-positive nuclei to total cell number is higher in day 7 than in day 8 vitrified in vitro-produced bovine embryos. AB - The aim of the present study was to find a reliable functional criterion for the evaluation of the proliferation potential of bovine in vitro-produced embryos. We used immunocytochemical detection of proliferating cell nuclear antigen (PCNA) combined with propidium iodide (PI) staining and subsequent confocal laser scanning microscopy together with routine morphological evaluation under a stereomicroscope to study fresh Day 7, 8, and 9, and cryopreserved Day 7 and 8 embryos. The ratio of PCNA/PI-positive nuclei was equal in fresh Day 7 and Day 8 embryos and significantly lower in Day 9 embryos. In general, Day 7 embryos tolerated the cryopreservation treatments better than Day 8 embryos. Vitrification in normal straws was especially detrimental to Day 8 embryos. In fresh Day 7 and 8 embryos, the PCNA results were in agreement with stereomicroscopic evaluation. However, in Day 9 fresh and in Day 7 and 8 treated embryos, the missing PCNA revealed disorders that were not observed under morphological evaluation. PCNA immunocytochemistry is an effective method to obtain information about the functional state of nuclei. The ratio of PCNA positive nuclei can provide more information and numerical data about the developmental potential of bovine embryos after cryopreservation. PMID- 11420223 TI - Estrogen receptor-alpha protein localization in the testis of the rainbow trout (Oncorhynchus mykiss) during different stages of the reproductive cycle. AB - Estrogen receptor-alpha (ER-alpha) is important for male reproduction in mammals; however, no information is available on ER-alpha protein distribution in the testes of fishes. The cellular localization of the rainbow trout (Oncorhynchus mykiss) ER-alpha (rtER-alpha) protein, throughout the annual reproductive cycle was determined in this study. An antibody was designed based on a 15-amino acid sequence from the D-domain of the rtER-alpha, and its specificity was confirmed using Western blot analysis. Immunohistochemical analysis revealed rtER-alpha protein to be present only in the testicular interstitium, at every stage of the annual reproductive cycle. The localization of rtER-alpha protein in the interstitial fibroblasts, the Leydig cell precursor in the rainbow trout, suggests a role for estrogens in the differentiation of these precursor cells into mature Leydig cells. This is the first study to report the cellular localization of an estrogen receptor protein in the testis of any fish species. PMID- 11420224 TI - Inhibitory action of leptin on early follicular growth differs in immature and adult female mice. AB - In order to investigate the action of leptin on early follicular growth, preantral follicles, 95-115 microm in diameter were mechanically isolated from the ovaries of BDF1 hybrid immature (11-day-old) and adult (8-wk-old) mice, and cultured for 4 days in vitro. Follicular growth was assessed by daily changes in follicular diameter and by the amount of estradiol and immunoreactive (IR) inhibin released into the culture medium at Day 4. Preantral follicles from immature mice showed a significant development in follicular growth as a result of stimulation by GH (1 mIU/ml), insulin-like growth factor (IGF)-I (100 ng/ml) + FSH (100 mIU/ml), and GH (1 mIU/ml) + FSH (100 mIU/ml). Although leptin at concentrations of 1-1000 ng/ml did not have any significant effect on follicular growth stimulated by IGF-I or GH, it significantly inhibited follicular growth in a dose-related manner when follicles were stimulated by IGF-I + FSH and GH + FSH, respectively, suggesting that leptin attenuated the additive effect of FSH. On the other hand, preantral follicles from adult mice were cultured in the presence of FSH, and FSH-dependent follicular growth was inhibited by leptin in a dose related manner. Because FSH stimulates cAMP production, we investigated the involvement of cAMP in the inhibitory mechanisms of leptin. Preantral follicles from immature and adult mice were cultured in the presence of either 8-Br-cAMP or forskolin. Both 8-Br-cAMP and forskolin significantly increased follicular diameter and hormone secretion in both immature and adult mice. However, 8-Br cAMP and forskolin-stimulated follicle growth and hormone secretion were significantly inhibited in immature mice by coadministration of leptin, whereas growth of preantral follicles from adult mice was not inhibited by addition of leptin to cultures. These results indicate that leptin causes an inhibitory effect on the early follicular development of both immature and adult mice, but the inhibitory mechanisms of leptin are different. PMID- 11420225 TI - 3alpha-hydroxysteroid dehydrogenase messenger RNA transcription in the immature rat ovary in response to an ovulatory dose of gonadotropin. AB - The ovulatory process in mammals involves a substantial increase in the metabolism of steroids and eicosanoids in response to a surge in LH or to an injection of hCG into experimental animals. This study provides evidence that the ovulatory stimulus causes induction of the gene for 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD), an enzyme that belongs to several oxidoreductase superfamilies that affect steroid and eicosanoid metabolism. Immature Wistar rats were primed with 10 IU eCG s.c., and 48 h later the 12-h ovulatory process was initiated by 10 IU hCG s.c. Ovarian RNA was extracted at 0, 2, 4, 8, 12, and 24 h after injecting the animals with hCG. The RNA extracts were used for reverse transcription-polymerase chain reaction (PCR) differential display to detect gene expression in the stimulated ovarian tissue. One of the PCR primer sets differentially amplified a cDNA fragment that is 52.3% homologous with a 3alpha HSD gene in rat liver. Northern analyses revealed that maximum transcription was at 8 h after the animals had been treated with hCG. The Northerns also indicated that the 3alpha-HSD cDNA probe cross-hybridized with as many as six different bands of mRNA on the blots. In situ hybridization localized 3alpha-HSD mRNA in the granulosa and thecal layers of mature follicles and in newly formed corpora lutea at 24 h after the ovulatory stimulus. In conclusion, gene(s) for 3alpha-HSD are transcribed in ovarian follicles in response to an ovulatory dose of gonadotropin. A possible function of the oxidoreductase enzyme that is translated from the 3alpha-HSD mRNA may be to reduce the toxic aldehyde and ketone components of the steroids and eicosanoids that accumulate in the mammalian ovary at the time of ovulation. PMID- 11420226 TI - Expression of the hamster sperm protein P26h during spermatogenesis. AB - P26h is a hamster sperm protein of 26 kDa that has been previously characterized as a surface protein covering the acrosome acquired during epididymal transit. P26h is involved in sperm-egg interactions. Recently, it has been shown that the P26h transcript is highly expressed in the testis, and the P26h cDNA has been cloned from a hamster testicular cDNA library. Herein we report the production of a fusion protein (maltose binding protein-P26h) with the whole P26h cDNA encoding sequence and the production of a polyclonal antiserum against it. In Western blots, this antiserum recognized both the P26h extracted from cauda epididymal spermatozoa and the MBP-P26h. We also determined the age of appearance of P26h and which germ cell types express P26h mRNA and its translational product. Northern blots and in situ hybridization analysis showed that P26h transcripts appear at 3 wk of age, within the first round of spermatogenesis in the golden hamster. In situ hybridization showed that P26h transcripts are expressed in spermatocytes and round spermatids, whereas immunostaining revealed the presence of P26h in the cytoplasm of round spermatids and elongated spermatids. P26h was undetectable in testicular spermatozoa. Both in situ hybridization and immunostaining showed P26h expression to be dependent of the testicular cell type and the epithelium cycle. The implications for P26h in sperm-egg interaction and the testicular origin of P26h are discussed. PMID- 11420227 TI - Apoptosis induced in rats by 4-vinylcyclohexene diepoxide is associated with activation of the caspase cascades. AB - Previous studies have shown that ovotoxicity induced in rats by dosing with 4 vinylcyclohexene diepoxide (VCD) is likely via acceleration of the normal rate of atresia (apoptosis). The present study was designed to investigate the apoptosis related caspase cascades as a component of this phenomenon in isolated ovarian small follicles. Female F344 rats were given a single dose of VCD (80 mg/kg, i.p., on Day 1; a time when ovotoxicity has not been initiated), or dosed daily for 15 days (80 mg/kg, i.p., on Day 15; a time when significant ovotoxicity is underway). Ovaries were collected after the final dose. Small preantral follicles (25-100 microm in diameter) were isolated, cellular fractions were prepared, and cleavage activity or protein expression levels of caspases-3, -8, and -9 were measured. Cytosolic caspase-3 activity was increased in small follicles (P < 0.01) by VCD treatment (Day 1, 2.86 +/- 0.23; Day 15, 3.25 +/- 0.64, VCD/control, n = 3). This activation was not seen in large or antral follicles (not targeted by VCD). Procaspase-3 protein was increased(P < 0.05) by VCD treatment 212% over controls in small ovarian follicles in Day 15, but not Day 1-dosed rats. Immunofluorescence staining intensity was evaluated by confocal microscopy. Caspase-3 protein, located in the cytosolic compartment of oocytes and granulosa cells of preantral follicles in various stages of development, was selectively increased (P < 0.05) in primordial and small primary follicles from Day 15 VCD dosed rats. Caspase-8 activity was increased in small follicles in Day 15, but not in Day 1-treated rats; whereas caspase-9 activity was increased by VCD on Day 1 in the mitochondrial fraction. Thus, these data provide evidence that accelerated atresia induced in small ovarian follicles in rats by VCD is associated with activation of a caspase-mediated cascade. PMID- 11420228 TI - Characterization of a putative membrane receptor for progesterone in rat granulosa cells. AB - Progesterone (P(4)) inhibits granulosa cell apoptosis in a steroid-specific, dose dependent manner, but these cells do not express the classic nuclear P(4) receptor. It has been proposed that P(4) mediates its action through a 60-kDa protein that functions as a membrane receptor. The present studies were designed to determine the P(4) binding characteristics of this protein. Western blot analysis using an antibody that recognizes the P(4) binding site of the nuclear P(4) receptor (C-262) confirmed that the 60-kDa protein was localized to the plasma membrane of both granulosa cells and spontaneously immortalized granulosa cells (SIGCs). To determine whether this protein binds P(4), proteins were immunoprecipitated with the C-262 antibody, electrophoresed, transferred to nitrocellulose, and probed with a horseradish peroxidase-labeled P(4) in the presence or absence of nonlabeled P(4). This study demonstrated that the 60-kDa protein specifically binds P(4). Scatchard plot analysis revealed that (3)H-P(4) binds to a single site (i.e., single protein), which is relatively abundant (200 pmol/mg) with a K(d) of 360 nM. (3)H-P(4) binding was not reduced by dexamethasone, mifepristone (RU 486), or onapristone (ZK98299). Further studies with SIGCs showed that P(4) inhibited apoptosis and mitogen-activated protein kinase kinase (MEK) activity, and maintained calcium homeostasis. These studies taken together support the concept that the 60-kDa P(4) binding protein functions as a low-affinity, high-capacity membrane receptor for P(4). PMID- 11420229 TI - A potential role for insulin-like growth factor binding protein-4 proteolysis in the establishment of ovarian follicular dominance in cattle. AB - A critical transition in ovarian follicular development is the selection of a dominant follicle, capable of ovulating, from a cohort of synchronously growing antral follicles. However, little is known about mechanisms and factors that regulate the selection and growth of dominant ovarian follicles. We have investigated whether a component of the insulin-like growth factor (IGF) system, namely IGFBP-4 protease, is associated with the establishment of follicular dominance in cattle. IGFBP proteases degrade IGFBPs, freeing IGFs to interact with their receptors. In experiment 1, follicular fluid from preovulatory follicles (n = 4) degraded about 80% of the added recombinant human (rh) IGFBP-4 within 18 h of incubation. The IGFBP-4 protease exhibited optimal activity at neutral/basic pH and its sensitivity to various protease inhibitors suggested a metalloprotease. The decline in the intensity of the band corresponding to intact rhIGFBP-4 was accompanied by the appearance of immunoreactive fragments of molecular weights approximately 18 and 14 kDa, which were not detectable by ligand blot analysis. In experiment 2, follicular fluid samples were collected from dominant and subordinate follicles on Day 2 or 3 of the first follicular wave, after ovariectomy (experiment 2a, n = 3/day) or by ultrasound-guided follicular aspiration (experiment 2b, n = 4-5/day). Estradiol concentrations in follicular fluid from dominant vs. subordinate follicles confirmed their identities and indicated that the dominant follicle had been selected by Day 2 of the follicular wave. In both experiments 2a and 2b, IGFBP-4 proteolytic activity was 2- to 3.5-fold (P < 0.05) and 5-fold (P < 0.01) higher in follicular fluid from dominant than subordinate follicles on Days 2 and 3 of the follicular wave, respectively. The finding that IGFBP-4 proteolytic activity is higher in dominant, estrogen-active follicles than in subordinate follicles of the same cohort, as early as Day 2 of the follicular wave, strongly suggests a role for IGFBP-4 protease in the establishment of ovarian follicular dominance. PMID- 11420230 TI - Development of codominant follicles in cattle is associated with a follicle stimulating hormone-dependent insulin-like growth factor binding protein-4 protease. AB - Low molecular weight insulin-like growth factor binding proteins (IGFBPs), particularly IGFBP-4, are believed to inhibit the actions of insulin-like growth factors (IGFs). We showed previously that ovarian follicular dominance in cattle is associated with the presence of a protease that degrades IGFBP-4. To test the hypothesis that specific IGFBP-4 proteolysis is associated with selection of the dominant follicle, we induced codominant follicles (co-DFs) during the first follicular wave of the estrous cycle. The ovaries of Holstein heifers were examined twice daily by ultrasonography; when the largest follicle reached 6 mm in diameter, saline (control, n = 5) or 2 mg of recombinant bovine (rb) FSH (FSH, n = 5) was injected i.m. every 12 h for 48 h. Follicular fluid was collected by aspiration from the two largest follicles/heifer 12 h after the last injection. IGFBPs in follicular fluid were quantified by Western ligand blotting/phosphorimaging. IGFBP-4 protease activity was measured by incubating follicular fluid with recombinant human (rh) IGFBP-4 substrate, followed by ligand blotting/phosphorimaging to quantify the percent of substrate loss and Western immunoblotting to detect specific proteolytic fragments. Co-DFs of FSH heifers did not differ (P > 0.05) from the single dominant follicle of controls in size, or in concentration of progesterone or level of IGFBP-4 in follicular fluid. In contrast, the largest subordinate follicle of control heifers was smaller, with lower progesterone and higher IGFBP-4 in the follicular fluid (P < 0.05). Concentrations of estradiol in follicular fluid were high in dominant follicles, intermediate in co-DFs, and low in subordinate follicles (P < 0.05). IGFBP-4 protease activity in co-DFs was similar (P > 0.05) to that of dominant follicles, but fourfold higher (P < 0.05) than that of subordinate follicles. The results strongly suggest that an FSH-dependent IGFBP-4 protease is associated with selection of the dominant follicle in cattle. PMID- 11420231 TI - Expression of stress response genes in germ cells during spermatogenesis. AB - During germ cell development different spermatogenic cell types show remarkable variation in their susceptibility to stressful stimuli. Various cellular mechanisms are triggered in germ cells after exposure to stress, but the expression of only a few of the genes involved in such pathways has been studied during spermatogenesis. In the present study we determined the expression profiles of 216 stress response genes in isolated rat germ cells (pachytene spermatocytes, and round and elongating spermatids) using cDNA atlas arrays. Of the 216 genes studied, 86 were detected in pachytene spermatocytes, 82 in round spermatids, and 52 in elongating spermatids. Fifty percent (48) of the total number of genes detected during spermatogenesis were detected in all three cell types while nearly 25% (25) were expressed exclusively in pachytene spermatocytes and round spermatids; some cell specific transcripts were observed also. The use of the K means clustering method allowed us to group genes by their pattern of expression during spermatogenesis; five specific expression profiles were obtained and analyzed. To determine how stress response genes are regulated throughout spermatogenesis, we examined the expression of genes involved in stress response mechanisms such as heat shock proteins-chaperones, DNA repair, and oxidative stress. Genes belonging to these families were differentially expressed during germ cell development. We suggest that the differential expression of stress response genes during spermatogenesis contributes to the selectivity of the susceptibility of germ cells to stress. PMID- 11420232 TI - Estrogen receptor beta in the sheep ovary during the estrous cycle and early pregnancy. AB - Objectives were to sequence and examine the expression of the estrogen receptor beta (ERbeta) in the sheep ovary. The sequence of the ovine ERbeta (oERbeta) was determined using reverse-transcription polymerase chain reaction (RT-PCR) and cloning techniques. The reading frame of oERbeta contained 527 amino acids and exhibited high overall homology with cow (98%), rat (88%), and human (88%) ERbeta. In addition, an oERbeta isoform having a 139-base pair deletion (oERbeta1) was identified. The predicted amino acid sequence of this isoform is lacking the ligand-binding and carboxyl-terminal transactivation domains. The oERbeta protein and mRNA were determined in ovaries obtained from ewes on Days 0 (first day of estrus), 2, 6, and 10 of the estrous cycle and Day 30 of gestation. Immunohistochemistry showed that oERbeta protein was located in granulosa cells, the ovarian surface epithelium, endothelium, and Day 2 corpus luteum (CL). Weak immunostaining for ERbeta was detected in the theca interna. Relative steady state amounts of oERbeta mRNA in the CL were determined using semiquantitative RT PCR. Amounts of oERbeta mRNA were greater (P < 0.05) during CL formation (Day 2) than at later stages. The oERbeta to oERbeta1 mRNA ratio was lower (P < 0.05) on Day 2 than on Day 10 or Day 30 due to a decrease in amounts of oERbeta1. Results indicate that the oERbeta is a 527-amino acid protein expressed in specific cells of the ovary. Changes in relative amounts of full-length oERB and a deletion isoform in CL occurred during the estrous cycle, suggesting that these two types of ERbeta might regulate estrogen actions during early CL development in sheep. PMID- 11420233 TI - Cyclooxygenase-2 and prostaglandin E(2)(PGE(2)) receptor messenger RNAs are affected by bovine oocyte maturation time and cumulus-oocyte complex quality, and PGE(2) induces moderate expansion of the bovine cumulus in vitro. AB - Expression of cyclooxygenase-2 (COX-2) and prostaglandin E(2) (PGE(2)) receptor 2 (EP2) are necessary for rodent cumulus expansion in vivo. Prostaglandin E(2) receptor 3 (EP3) has been detected in bovine preovulatory follicles and corpora lutea. The current experiments examined the effect of PGE(2) on bovine cumulus expansion in vitro and expression of COX-2, EP1, EP2, EP3, and EP4 mRNAs in bovine cumulus-oocyte complexes (COCs) at 0, 6, 12, 18, and 24 h time points during maturation in vitro. Concentrations of PGE(2) above 50 ng/ml resulted in moderate cumulus expansion of bovine COCs, but expansion did not occur in the absence of serum. COX-2 mRNA expression increased in bovine COCs at 6 h and 12 h of maturation, then decreased. EP2 mRNA was detectable by reverse transcription polymerase chain reaction at all time points. EP3 mRNA expression increased in COCs from 0 to 6 h and remained at this higher level through the culture period. Very low levels of EP4 mRNA expression were detectable, but EP1 was not detected in bovine COCs. Because EP receptor mRNAs and COX-2 mRNA are expressed in bovine COCs, there exists the potential for a prostaglandin autocrine/paracrine regulatory pathway during oocyte maturation. Differential expression of the EP3 mRNA among varying COC classes indicates that this gene product may be a useful marker of oocyte competence. Although the PGE(2) pathway is involved in cumulus expansion, serum factors are required to mediate PGE(2)-induced expansion. PMID- 11420234 TI - Cellular and morphological traits of oocytes retrieved from aging mice after exogenous ovarian stimulation. AB - The present study aims to shed light on the origin of abnormal oocytes ovulated by aged females. In order to reach this goal, cellular and morphological traits of ovulated oocytes from hybrid (C57Bl/6JIco female x CBA/JIco male) female mice retrieved after exogenous ovarian stimulation at the age of 12, 40-42, 50-52, or 57-62 wk were analyzed. Aging of female mice was associated with 1) decreased number of ovulated oocytes; 2) increased percentage of cumulus-free oocytes; 3) raised percentage of oocytes with intracellular mitochondrial aggregates; 4) reduced percentage of oocytes displaying a normal distribution of chromosomes in the metaphase-II plate; 5) increased percentage of normal oocytes exhibiting a DNA-containing polar body (PB); 6) higher percentage of oocytes with chromosome scattering; 7) increased percentage of chromosome-scattered oocytes without a DNA containing PB and with intracytoplasmic mitochondrial aggregates; 8) raised percentage of oocytes exhibiting chromosome decondensation; 9) lower percentage of chromosome-decondensed oocytes lacking both a DNA-containing PB and intracytoplasmic mitochondrial aggregates; 10) increased percentage of abnormal/degenerated oocytes; 11) reduced percentage of abnormal/degenerated oocytes displaying cellular fragmentation; and 12) higher percentage of abnormal/degenerated oocytes with mitochondrial aggregates exhibiting no nuclear/chromosomal DNA fluorescence, cellular fragmentation, milky or dark cytoplasm, or cellular remains enclosed by the zona pellucida. Although several studies suggest aging females may ovulate aged or overripened oocytes, these data support the hypothesis that old females ovulate an increased percentage of atretic/apoptotic oocytes coming from rescued follicles that would have become atretic earlier in life. PMID- 11420235 TI - Differential expression of the C(s) and Calpha1 isoforms of the catalytic subunit of cyclic 3',5'-adenosine monophosphate-dependent protein kinase testicular cells. AB - The amino terminus of the sperm cAMP-dependent protein kinase catalytic subunit (termed C(s)) differs from that of the Calpha1 isoform expressed in most tissues due to the use of alternative transcripts of the Calpha gene. Both Calpha1 and C(s) transcripts are present in testis; C(s) is expressed specifically in spermatogenic cells and is the only C isoform detected in mature sperm. Immunohistochemistry of mouse testis using antibodies specific for C(s) and Calpha1 now shows that Calpha1 is present in somatic testicular cells, spermatogonia, and preleptotene spermatocytes but not in cells that are in later stages of spermatogenesis. In contrast, C(s) is expressed only in midpachytene and later stage spermatocytes and in spermatids. Therefore, C(s) and Calpha1 expression do not overlap. Immunofluorescence microscopic localization of C(s) in murine and ovine sperm reveals that C(s) is located primarily in sperm tail components, including the midpiece mitochondria and the axoneme. Quantitative analysis of Western blots indicates that individual ovine sperm contain approximately 4 x 10(5) molecules of C(s), a seemingly large number for a protein that acts catalytically. PMID- 11420236 TI - Lupron depot prevention of antispermatogenic/antifertility activity of the indenopyridine, CDB-4022, in the rat. AB - The goals of this study were to determine the CDB-4022 dose-response relationship for induction of acute decreases in testicular weight and germ cell depopulation in rats; establish the threshold dose of CDB-4022 required to induce infertility; and investigate whether CDB-4022-induced testicular damage could be prevented by a GnRH agonist (Lupron Depot). Reduction of testis weight and germ cell depopulation were observed 7 days after a single oral dose of 1 mg CDB-4022/kg, whereas 0.5 mg/kg had no observable effect. These effects were maximal at 12.5 or 25 mg CDB-4022/kg. After a single oral dose of either 2.5 or 5 mg/kg, CDB-4022 induced infertility in five of five treated rats by Week 5, whereas only one of five males was rendered infertile at a dose of 1 mg/kg. Proven fertile male rats (6/group) were treated with vehicle, CDB-4022 alone (2.5 mg/kg on Day 0), CDB 4022 plus Lupron Depot (on Weeks -1, 2, 5, and 8), or Lupron Depot alone. Control males demonstrated normal fertility throughout a 32-wk cohabitation period. Five of six rats were rendered transiently infertile with Lupron Depot alone, but all recovered fertility. CDB-4022 treatment resulted in infertility in all six rats, and only one of six regained fertility. Combined treatment also caused infertility in all six rats, but four of six recovered fertility (P = 0.08 compared to CDB-4022 alone). Testicular weight was decreased in the three treatment groups compared to vehicle controls; testicular weights were ranked from highest to lowest as follows: vehicle > Lupron Depot > Lupron Depot + CDB 4022 > CDB-4022. The tubule differentiation index of Lupron Depot-treated rats (96 +/- 4%) was not different from vehicle-treated rats (100%). CDB-4022 treatment decreased the number of differentiating tubules (15 +/- 8%). Lupron Depot plus CDB-4022 treatment resulted in a greater number of differentiating tubules (53 +/- 12%) than CDB-4022 alone, but this was still lower than vehicle- or Lupron Depot-treated rats. These data indicate that 2.5 mg/kg of CDB-4022 was the oral threshold dose that caused testicular damage rendering the majority of adult male rats permanently infertile within the study interval; 12.5 mg/kg of CDB-4022 induced maximal testicular damage. Suppression of gonadotropins and/or testosterone production by treatment with Lupron Depot before and after CDB-4022 prevented the CDB-4022-induced irreversible testicular damage. PMID- 11420237 TI - Gonadotropin-releasing hormone content in the brain and pituitary of male and female grass rockfish (Sebastes rastrelliger) in relation to seasonal changes in reproductive status. AB - The involvement of individual molecular forms of GnRH in the regulation of reproductive cyclicity in a viviparous marine teleost, the grass rockfish (Sebastes rastrelliger), was evaluated by relating the brain and pituitary content of the neuropeptide to reproductive status. The presence of sea bream (sb) GnRH, chicken GnRH-II, and salmon GnRH in the brain was confirmed by their elution pattern on HPLC and RIA. In addition, HPLC elution profiles suggest that there may be a fourth form of GnRH. All forms of GnRH were found in male and female brains in all reproductive conditions. However, only sbGnRH could be detected in appreciable amounts in the pituitary. Of the four forms of GnRH found in the rockfish, only sbGnRH fluctuated during the reproductive cycle and large accumulations were detected in the brains and pituitaries of postspawn females and regressed males. The accumulation of sbGnRH at the end of the reproductive cycle is suggested to reflect a decline in GnRH secretion relative to synthesis. The dominance of sbGnRH in the pituitary and its individual fluctuation in relation to seasonal changes in reproductive status suggests that sbGnRH is an important regulator of gonadotropin-mediated reproductive activity in rockfish. PMID- 11420238 TI - Lysophosphatidic acid antagonizes the morphoregulatory effects of the luteinizing hormone on luteal cells: possible role of small Rho-G-proteins. AB - Lysophosphatidic acid (LPA) is a biologically active phospholipid recently introduced as a new marker for ovarian cancer. Because high concentrations of LPA have also been found in the follicular fluid from healthy subjects, one can presume that this biological mediator may have relevance for normal ovarian physiology as well. We have reported earlier that luteal cells possess specific binding sites for LPA. Using these cells as a model, we show now that LPA is able to modulate the morphological cell shape changes induced by LH in that it inhibits the formation of stellate processes induced by LH. This morphoregulatory effect of LPA is mimicked by cytotoxic necrotizing factor 1, a bacterial toxin known to activate small G-proteins from the Rho family. On the other hand, C3 exotransferase that acts mainly through the inhibition of Rho A mimics the effects of LH. Furthermore, we report here that the morphoregulatory effects of LPA are accompanied by the translocation of Rho proteins from the cytosol to cell membrane, an effect generally considered to be an indicator for the activation of Rho-GTPases. During the development and rescue of the corpus luteum, major morphoregulatory effects are exerted by LH that appear to be modulated by LPA via an activation of Rho proteins. PMID- 11420239 TI - Cloning and characterization of the cyclic guanosine monophosphate-inhibited phosphodiesterase PDE3A expressed in mouse oocyte. AB - In the preovulatory follicle, oocyte meiotic resumption occurs soon after the LH surge and is associated with a decrease in cAMP. Inhibition of cAMP degradation blocks germinal vesicle breakdown as well as activation of meiotic promoting factor, both hallmarks of reentry into the cell cycle. In situ and pharmacological analysis of rodent ovaries suggested the presence of a phosphodiesterase 3 (PDE3) in the germ cell but not the somatic cell compartment. Here we have investigated the structure and properties of the PDE form expressed in mouse oocytes. Polymerase chain reactions using a mouse oocyte cDNA library as a template, and primers based on the conserved sequence of rat and human PDE3As, yielded partial fragments corresponding to mouse PDE3A. Further screening of the mouse oocyte cDNA library and subsequent ligation of individual cDNA clones yielded PDE3A cDNA containing the entire coding region of mouse PDE3A. To determine the kinetic properties of this PDE, the cDNAs encoding the full-length PDE3A and NH(2)-truncation forms Delta 1 (Delta346aa) and Delta 2 (Delta608aa) were expressed in mouse Leydig tumor cells. Whereas the full-length recombinant protein was always found in the particulate fraction, the Delta 1 and Delta 2 truncated PDE3As were recovered mostly in the soluble fraction. The Michaelis constant values for hydrolysis of cAMP of PDE3A Delta 1 and PDE3A Delta 2 were similar to those of intact full-length PDE3A or oocyte PDE (0.2-0.5 microM). More importantly, there was good correlation between the rank of potency of selective and nonselective compounds in inhibiting recombinant PDE3A or PDE activity derived from cumulus-oocyte complexes and in blocking resumption of meiosis. These data provide evidence that the PDE expressed in the oocyte is a soluble form of PDE3A and that activity of this enzyme is involved in the control of resumption of meiosis. PMID- 11420240 TI - Epidermal growth factor regulation of equine glycoprotein hormone alpha subunit expression in trophoblast cells. AB - Primates and equids are the only species known to express the placental glycoprotein hormone, chorionic gonadotropin (CG), a heterodimeric glycoprotein composed of an alpha subunit linked to a hormone-specific beta subunit. The regulatory mechanisms involved in the induction of equine glycoprotein alpha subunit gene expression have not been identified. Epidermal growth factor (EGF) receptor is known to transduce signals that alter a number of different cellular functions (cell proliferation, differentiation, hormone secretion, and gene regulation). In the present study, we investigated the regulation of the equine alpha subunit gene by EGF in trophoblasts. We found that 2800 base pairs of 5' flanking sequence from the equine alpha subunit promoter is sufficient for basal expression in human choriocarcinoma cells. Epidermal growth factor and phorbol 12 myristate 13-acetate (PMA), an activator of protein kinase C (PKC), increased transcriptional activity of the equine alpha subunit promoter (-2800/+21). These responses were blocked by pretreatment with bisindolylmaleimide-I, an inhibitor of PKC, suggesting an involvement of this pathway downstream of EGF. In addition, PD98059, an inhibitor of the extracellular signal-regulated kinase (ERK) pathway, completely blocked activation of the equine alpha promoter by PMA, suggesting that mitogen-activated protein kinase (MAPK) cascade was involved downstream of the PKC pathway. In conclusion, the EGF/PKC/MAPK pathway regulates equine glycoprotein alpha subunit gene expression through a distinct regulatory region ( 2300 to -1900) in trophoblasts, while essential elements for basal expression appear to exist within the -2800 to -1900 region of the promoter. PMID- 11420241 TI - Chromosomal abnormalities and developmental kinetics in in vivo-developed cattle embryos at days 2 to 5 after ovulation. AB - The frequency of chromosome abnormalities was investigated in cattle embryos (n = 256) derived from superovulated heifers (n = 35) on Days 2, 3, 4, and 5 postovulation (PO). Interphase nuclei (n = 4358) were analyzed for chromosome abnormalities using fluorescent in situ hybridization with chromosome 6- and chromosome 7-specific probes and the developmental rate was described by scoring cell numbers. We found that 93%, 85%, 84%, and 69% of the embryos from Days 2, 3, 4, and 5 PO, respectively, displayed a normal diploid chromosome number in all cells. Of the embryos containing abnormal cells, mixoploidy was significantly more frequent than polyploidy. The percentage of mixoploidy at Days 2, 3, 4, and 5 PO was 5%, 13%, 16%, and 31%, respectively, whereas the percentages of polyploidy were 2%, 2%, 0%, and 0%, respectively. The mean number of cells per embryo was 4.7, 8, 11.5, and 48.3, respectively, at Days 2, 3, 4, and 5 PO. Thus, in vivo-developed embryos were significantly more advanced than the in vitro produced (IVP) embryos except for Day 2. In conclusion, a significantly lower frequency of chromosomally abnormal embryos, in particular displaying polyploidy early after fertilization, was seen in in vivo versus IVP embryos, and these chromosomal abnormalities may be inherent to the process of IVP in cattle. PMID- 11420242 TI - Production and endocrine role of inhibin during the early development of bull calves. AB - This study investigated the ontogeny of control of FSH secretion by inhibin during early prepubertal development of bulls by 1) measurements of circulating levels of inhibin and FSH from 1 to 13 wk of age, and 2) immunoneutralization of endogenous inhibin at 7, 21, 60, and 120 days of age. In addition, production and localization of inhibin in testes were examined by immunohistochemistry and Western blots at 7, 21, 60, and 120 days of age. Plasma immunoreactive inhibin levels were relatively low between 1 and 3 wk of age and then showed a tendency to rise (P < 0.1) from 4 wk of age. Circulating concentrations of FSH were low during 3 wk after birth and increased at 5 wk, remained high (P < 0.05) until 16 wk of age. Treatment with inhibin antiserum resulted in a significant (P < 0.05) increase in plasma FSH at 7, 21, 60, and 120 days of age compared to those following injection of control serum; however, the magnitude of the FSH rise after inhibin immunization was greater as bulls aged. There were no significant changes in plasma LH after inhibin immunization. An intense staining of inhibin alpha subunits was found in Sertoli cells within the solid seminiferous cords from 7 to 120 days of age, while no specific immune reaction was found in interstitial cells. Western blot analysis of testicular homogenates isolated from bulls 7-120 days of age revealed presence of a 28.5-kDa molecule that cross reacted with inhibin alpha subunit and beta(B) subunit-specific antibodies. In this study, before 13 wk of age in bull calves, there was no inverse relationship between plasma concentrations of immunoreactive inhibin and FSH. However, the present immunization study clearly indicates that inhibin participates in the regulation of FSH secretion from infancy to early prepubertal stage, although the endocrine significance of inhibin becomes greater in older bulls. The results also indicate that the major production site of inhibin in the testis is Sertoli cells and that these cells produce inhibin that exerts a negative feedback effect on FSH secretion from early stages of development. PMID- 11420243 TI - Ontogeny of steroidogenesis in the fetal sheep gonad. AB - The aim of this study was to determine 1) the time of onset and cellular localization of gene expression for steroidogenic factor-1 (SF-1), steroidogenic acute regulatory protein, 3beta-hydroxysteroid dehydrogenase/Delta(5),Delta(4) isomerase (3beta-HSD), and the cytochrome P450 enzymes for cholesterol side-chain cleavage (P450(scc)), 17alpha-hydroxylase (P450(17alphaOH)), and aromatase (P450(arom)) during gonadal development; and 2) the amount of progesterone, androstenedione, testosterone, and 17beta-estradiol present in the fetal sheep gonad. Fetuses were collected on Days 24, 26, 28, 30, 32, 35, 40, 55, and 75 of gestation, and gene expression was determined by in situ hybridization. The steroid content of gonads collected on Days 30, 35, 55, and 75 of gestation was determined by RIA. Developing gonads collected from both male and female fetuses were steroidogenically active around the time of morphological sexual differentiation. In the female, the steroidogenic cells were initially located at the boundary of the cortex and medulla but become increasingly restricted to the mesonephric-derived cell streams. In the male, once tubules were identifiable, steroidogenesis was restricted to the interstitial regions. Interestingly, expression of both SF-1 and 3beta-HSD was observed prior to morphological sexual differentiation. In addition, expression of both of these genes was more widespread than the other genes in both males and females. PMID- 11420244 TI - Effects of hyperthermia on spermatogenesis, apoptosis, gene expression, and fertility in adult male mice. AB - Testicular heat shock was used to characterize cellular and molecular mechanisms involved in male fertility. This model is relevant because heat shock proteins (HSPs) are required for spermatogenesis and also protect cells from environmental hazards such as heat, radiation, and chemicals. Cellular and molecular methods were used to characterize effects of testicular heat shock (43 degrees C for 20 min) at different times posttreatment. Mating studies confirmed conclusions, based on histopathology, that spermatocytes are the most susceptible cell type. Apoptosis in spermatocytes was confirmed by TUNEL, and was temporally correlated with the expression of stress-inducible Hsp70-1 and Hsp70-3 proteins in spermatocytes. To further characterize gene expression networks associated with heat shock-induced effects, we used DNA microarrays to interrogate the expression of 2208 genes and thousands more expression sequence tags expressed in mouse testis. Of these genes, 27 were up-regulated and 151 were down-regulated after heat shock. Array data were concordant with the disruption of meiotic spermatogenesis, the heat-induced expression of HSPs, and an increase in apoptotic spermatocytes. Furthermore, array data indicated increased expression of four additional non-HSP stress response genes, and eight cell-adhesion, signaling, and signal-transduction genes. Decreased expression was recorded for 10 DNA repair and recombination genes; 9 protein synthesis, folding, and targeting genes; 9 cell cycle genes; 5 apoptosis genes; and 4 glutathione metabolism genes. Thus, the array data identify numerous candidate genes for further analysis in the heat-shocked testis model, and suggest multiple possible mechanisms for heat shock-induced infertility. PMID- 11420245 TI - Effects of matrix proteins on the expression of matrix metalloproteinase-2, -9, and -14 and tissue inhibitors of metalloproteinases in human cytotrophoblast cells during the first trimester. AB - The activity of matrix metalloproteinases (MMPs) specifies the ability of the trophoblast cell to degrade extracellular matrix (ECM) substrates. Usually the process of normal human placentation involves a coordinated interaction between the fetal-derived trophoblast cells and their microenvironment in the uterus. In this study, the effects of ECM proteins on the expression of MMP-2, -9, and -14 (membrane-type MMP-1); and the production of tissue inhibitors of metalloproteinase (TIMP) types -1, -2, and -3 have been investigated. Cytotrophoblast cells at 9 or 10 wk of gestation were cultured on various ECM coated dishes under serum-free conditions. Gelatin zymography analysis showed that cells grown on fibronectin (FN), laminin (LN), and vitronectin (VN) secreted more MMP-9 (about 1.5- to 3-fold more) than cells cultured on collagen I (Col I), whereas the secretion of MMP-9 by cells cultured on collagen IV (Col IV) was only half that by the cells on Col I. Northern Blot analysis gave the same results as zymography, indicating that expression of the MMP-9 gene in cytotrophoblast cells can be affected by matrix proteins. There was no significant difference in the expression of MMP-2 either at protein or mRNA levels among the cells cultured on the different matrix substrates. The expression of MMP-14 was regulated in a manner similar to that of MMP-2. Using ELISA, we detected higher levels of TIMP-1 in the culture medium of cells grown on VN, LN, and FN compared with that grown on Col I. But the expression of TIMP-3 mRNA was remarkably inhibited by VN, and ECM proteins had no effect on TIMP-1 and TIMP-2 mRNA expression. It was also observed that cultured cytotrophoblast cells expressed the corresponding receptors for the tested matrix proteins, such as integrins alpha(1), alpha(5), alpha(6), beta(1), and beta(4). Furthermore, the adhesiveness of cytotrophoblast cells on Col I, Col IV, FN, and LN was increased by 62%, 45%, 21%, and 22%, respectively, when compared with adhesiveness on VN. Isolated cytotrophoblast cells remained stationary when cultured on dishes coated with Col I and Col IV, but they assumed a more motile morphology and aggregated into a network when cultured on LN and VN. These data indicate that human trophoblast cells interact with their microenvironment to control their behavior and function. PMID- 11420246 TI - Cell coupling and maturation-promoting factor activity in in vitro-matured prepubertal and adult sheep oocytes. AB - We examined some differences between prepubertal and adult ovine oocytes; in particular we analyzed the functional status of the cumulus-oocyte complex, protein synthesis during in vitro maturation, and because no information is available on prepubertal and adult sheep, maturation-promoting factor (MPF) fluctuations throughout meiotic progression both in prepubertal and adult sheep oocytes. After 24 h of maturation, percentages of MII oocytes were similar between prepubertal and adult animals. Electron microscopy examinations showed that prepubertal oocytes had fewer transzonal projections than adult oocytes. Methionine uptake was significantly lower in prepubertal cumulus-enclosed oocytes examined through meiotic progression. On the contrary, denuded prepubertal oocytes showed a higher methionine incorporation in the first 4 h of incubation compared with adult oocytes. We also found some differences in MPF activity between prepubertal and adult oocytes at MII stage. In fact, prepubertal MII oocytes had a significantly lower level of MPF activity than adult oocytes did and, after fusion with germinal vesicle oocytes, they were unable to induce nuclear breakdown and chromosome condensation 1-2 h post-fusion, whereas adult MII oocytes could induce these processes. Our findings show that the lesser competence of prepubertal oocytes could be due to morphological anomalies and alterations in physiological activity and that oocytes do not reach full developmental competence until puberty. PMID- 11420247 TI - Parthenogenetic activation of rhesus monkey oocytes and reconstructed embryos. AB - This study determines the efficiency of sequential calcium treatments (electroporation or ionomycin) combined with protein synthesis (cycloheximide) or phosphorylation inhibitors (6-dimethylaminopurine) or the specific maturation promoting factor (MPF) inhibitor, roscovitine, in inducing artificial activation and development of rhesus macaque parthenotes or nuclear transfer embryos. Exposure of oocytes arrested at metaphase II (MII) to ionomycin followed by 6 dimethylaminopurine or to electroporation followed by cycloheximide and cytochalasin B induced pronuclear formation and development to the blastocyst stage at a rate similar to control embryos produced by intracytoplasmic sperm injection. Parthenotes did not complete meiosis or extrude a second polar body, consistent with their presumed diploid status. In contrast, oocytes treated sequentially with ionomycin and roscovitine extruded the second polar body and formed a pronucleus at a rate higher than that observed in controls. Following reconstruction by nuclear transfer, activation with ionomycin/6 dimethylaminopurine resulted in embryos that contained a single pronucleus and no polar bodies. All nuclear transfer embryos activated with ionomycin/roscovitine contained one large pronucleus. However, a third of these embryos emitted one or two polar bodies, clearly containing chromatin material. In summary, we have identified simple yet effective methods of oocyte or cytoplast activation in the monkey, ionomycin/6-dimethylaminopurine, electroporation/cycloheximide/cytochalasin B, and ionomycin/roscovitine, which are applicable to parthenote or nuclear transfer embryo production. PMID- 11420248 TI - Interaction of endophyte-infected fescue and heat stress on ovarian function in the beef heifer. AB - The objective of the experiment was to examine the interaction of endophyte infected tall fescue and environmental temperature on follicular and luteal development and function in beef heifers. Heifers were fed endophyte-free or endophyte-infected tall fescue seed at thermoneutral or heat stress temperatures (n = 6/treatment) 4 wk before and 3 wk after synchronized ovulation. All heifers were subjected to thermoneutral conditions (19 degrees C, 50% relative humidity) from Days -7 to -2; temperature increased incrementally from Days -1 to 0 and cycled between 25 degrees C and 31 degrees C between Days 1 and 20 for heat stressed heifers. Serum was collected and ovaries monitored every other day after induced luteolysis between Days 1 and 23 or until ovulation. Size and location of follicles >4 mm and corpora lutea were recorded. Serum concentrations of prolactin were reduced in heat-stressed heifers fed infected seed and both heat stress and infected seed decreased total cholesterol. Rectal temperature and respiration rate were greatest in heifers fed the infected seed when exposed to maximal temperatures. Heat stress led to reduced diameter of the corpus luteum and serum progesterone compared with thermoneutral conditions. Progesterone was reduced more so in heifers fed infected seed. The combination of infected seed and heat stress was associated with reduced diameter of the preovulatory dominant follicle, and consumption of infected seed led to fewer large follicles during the estrous cycle. Both stressors led to reduced serum estradiol. Impaired follicle function may explain reduced pregnancy rates commonly observed in heifers grazing infected tall fescue pasture. PMID- 11420249 TI - DNA array analysis of changes in preovulatory gene expression in the rat ovary. AB - During the periovulatory period, the mammalian ovary is the site of dramatic functional and structural changes, leading to oocyte maturation, follicle rupture, and corpus luteum formation. To a large extent, these processes result from changes in the transcriptome of various ovarian cell types. To develop a broader view of periovulatory changes in gene expression in the ovary and to identify further genes involved in periovulatory events, we used the recently developed DNA array technology. Immature female eCG-primed rats were killed either immediately before or 6 h after ovulation induction with hCG. Total ovarian RNA was isolated and used to prepare radiolabeled cDNA probes, which were hybridized to DNA arrays representing approximately 600 rat genes. Quantitative analysis identified a multitude of regulated gene messages, including several genes involved in extracellular matrix degradation and lipid/steroid metabolism previously reported to be induced by hCG. This screening also identified a group of candidate genes whose ovarian expression and gonadotropin regulation was hitherto unknown. The induction of three of these genes, encoding cutaneous fatty acid-binding protein, the interleukin-4 receptor alpha chain, and prepronociceptin, was confirmed and further characterized by Northern blot analysis. In addition, in situ hybridization analysis showed that hCG administration resulted in exclusive or predominant expression of all three genes in theca cells. These results demonstrate that DNA arrays can be used to identify genes regulated during the periovulatory period, thus contributing to a more detailed understanding of the molecular mechanisms of ovulation. PMID- 11420250 TI - Developmental expression and characterization of FS39, a testis complementary DNA encoding an intermediate filament-related protein of the sperm fibrous sheath. AB - Proteins immunologically related to intermediate filaments have been identified in the sperm fibrous sheath but remain uncharacterized. We isolated and characterized a novel intermediate filament-related protein (FS39) localized to the fibrous sheath of the sperm tail. We used Northern blot analysis to establish that FS39 is transcribed predominantly in the testis of mice >18-20 days old. At this age, spermatogenesis has proceeded to the development of the first round haploid spermatids. In situ hybridization revealed that FS39 mRNA is first detectable in late step 3 spermatids, is at its highest level during steps 9 and 10, and diminishes in steps 13 and 14. Western blot analysis identified a single protein of 39 kDa in mouse and rat testis and epididymis, suggesting the protein is conserved in rodents. Indirect immunofluorescence localized FS39 to the fibrous sheath of the sperm tail, and in testis sections expression was detected from step 13 and step 14 spermatids onward, indicating FS39 is under translational control. Southern blot analysis showed FS39 to be a single copy gene, and hybridization to human genomic DNA suggested that a human equivalent gene is present. These results demonstrate that FS39 is transcribed in testis tissue during the haploid phase of spermatogenesis, is present in mature sperm, and codes for a novel 39-kDa intermediate filament-related protein of the fibrous sheath. PMID- 11420251 TI - Follicle-stimulating hormone and its alpha and beta subunits in rainbow trout (Oncorhynchus mykiss): purification, characterization, development of specific radioimmunoassays, and their seasonal plasma and pituitary concentrations in females. AB - Gonad development in fish, as in mammals, is regulated by two gonadotropins (GTHs), FSH and LH. The function of LH in fish has been clearly established; however, the function(s) of FSH is less certain. The lack of specific and sensitive assays to quantify FSH and its alpha and beta subunits has hindered studies to assess physiological function. In this study, gel filtration chromatography, ion exchange chromatography, and HPLC were employed to purify FSH and its subunits from pituitary glands of rainbow trout (Oncorhynchus mykiss), and the identities of the isolates were confirmed by amino acid analysis. Polyclonal antibodies were raised against the free GTHalpha2 and free FSHbeta subunits to develop specific RIAs. The sensitivities of the intact FSH, GTHalpha2, and FSHbeta assays were 1 ng/ml, 0.2 ng/ml, and 0.1 ng/ml, respectively, and the cross-reaction of these molecules with each other and with intact LH in the heterologous assays was <10.4% throughout. Pituitary and plasma samples diluted in parallel with the standards in all three assays and spiked sample recoveries were >90% throughout. Measurement of plasma and pituitary concentrations of intact FSH in female rainbow trout confirmed the established seasonal profiles. Concentrations of free GTHalpha2 subunit were elevated both in the plasma and in the pituitary in females at ovulation (maximum concentrations: 34.93 +/- 6.3 ng/ml in plasma; 37.63 +/- 5.79 microg/pituitary). In both the plasma and the pituitary, free FSHbeta subunit was present throughout the reproductive cycle but at very low concentrations when compared with both free GTHalpha2 and intact FSH. The presence of free GTHalpha2 subunit in the plasma similarly occurs in mammals, but its functional significance in fish has yet to be established. PMID- 11420252 TI - Physiological regulation of maternal behavior in heifers: roles of genital stimulation, intracerebral oxytocin release, and ovarian steroids. AB - We tested the hypotheses that 1) epidural anesthesia at parturition would block both peripheral and central release of oxytocin and eliminate the development of maternal behavior in primiparous heifers and 2) estradiol priming, genital stimulation, and appropriate neonatal stimuli would induce maternal behavior in nulliparous heifers. In experiment 1, primiparous crossbred heifers (n = 13) with cannulas in the third cerebroventricle (IIIV) were assigned randomly to receive epidural treatments of saline (SAL; n = 6) or lidocaine HCl (EPI; n = 7) at the onset of labor induced between Days 270 and 280 of gestation. Epidural anesthesia blocked (P < 0.001) both central and peripheral release of oxytocin and markedly reduced (P < 0.05) or eliminated licking behaviors during a 3-h period following parturition as compared with SAL. Following approximately 1 wk of controlled daily suckling, during which calves were permitted access only to the inguinal region of their dams (three times daily for 10 min each time), a second maternal behavior test was performed. Although licking behavior remained markedly reduced (P < 0.001) in the EPI compared with the SAL groups, all heifers accepted their calf at the udder. In experiments 2-4, neither estradiol priming in ovariectomized heifers nor estradiol plus progesterone in intact heifers resulted in an induction of maternal behaviors following genital stimulation and presentation of a neonate wetted with amniotic fluid. Pelvic sensory deficits apparently block oxytocin release and disturb both short-latency and long-term maternal behaviors but do not result ultimately in rejection of the calf. Combinations of hormonal, sensory, olfactory, and visual cues observed previously to induce maternal behavior in nulliparous ewes do not appear adequate for induction of maternal behavior in nulliparous heifers. PMID- 11420253 TI - TSG-6 is concentrated in the extracellular matrix of mouse cumulus oocyte complexes through hyaluronan and inter-alpha-inhibitor binding. AB - During development of ovarian follicles in mammals, cumulus cells and the oocyte form a mucoelastic mass that detaches itself from peripheral granulosa cell layers upon an ovulatory surge. The integrity of this cumulus-oocyte complex (COC) relies on the cohesiveness of a hyaluronan (HA)-enriched extracellular matrix (ECM). We previously identified a serum glycoprotein, inter-alpha inhibitor (IalphaI), that is critical in organizing and stabilizing this matrix. Following an ovulatory stimulus, IalphaI diffuses into the follicular fluid and becomes integrated in the ECM through its association with HA. TSG-6 (the secreted product of the tumor necrosis factor-stimulated gene 6), another HA binding protein, forms a complex with IalphaI in synovial fluid. The purpose of this study was to investigate whether TSG-6 is involved in the ECM organization of COCs. Immunolocalization of TSG-6 and IalphaI in mouse COCs at different ovulatory stages was analyzed by immunofluorescence and laser confocal microscopy. IalphaI, TSG-6, and HA colocolized in the cumulus ECM. Western blot analyses were consistent with the presence of both TSG-6 and TSG-6/IalphaI complexes in ovulated COCs. These results suggest that TSG-6 has a structural role in COC matrix formation possibly mediating cross-linking of separate HA molecules through its binding to IalphaI. PMID- 11420254 TI - Nuclear transfer protocol affects messenger RNA expression patterns in cloned bovine blastocysts. AB - The successful production of embryos by nuclear transfer (NT) employing cultured somatic donor cells depends upon a variety of factors. The objective of the present study was to investigate the effects 1) of two different activation protocols, 2) the use of quiescent or nonquiescent donor cells (G(0) or G(1) of the cell cycle), and 3) passage number of donor cells on the relative abundance (RA) of eight specific mRNAs (DNA methyltransferase, DNMT; mammalian achaete scute homologue, Mash2; glucose transporter-1, Glut-1; heat shock protein 70.1, Hsp; desmocollin II, Dc II; E-cadherin, E-cad; interferon tau, IF; insulin-like growth factor 2 receptor, Igf2r) in single blastocysts employing a semiquantitative reverse transcription-polymerase chain reaction assay. The results were compared with those for their in vitro (IVP)- and in vivo-generated noncloned counterparts. In experiment 1, employing either FBA (fusion before activation) or AFS (fusion and activation simultaneously) to generate NT blastocysts, Hsp mRNAs were not found in NT embryos from either protocol, whereas Hsp transcripts were detectable in IVP embryos. The relative abundance (RA) of IF transcripts was significantly increased in the AFS and IVP groups compared to the FBA treatment. In experiment 2, the use of either G(0) or G(1) donor cells to produce cloned embryos both significantly reduced the relative amount of DNMT transcripts and significantly increased the RA of Mash2 compared to the IVP embryos. In addition, IF transcript levels were significantly elevated in NT blastocysts employing G(1) donor cells for NT compared to IVP embryos and those generated using G(0) cells. In experiment 3, donor cells, either from passsage 5/6 or 8, were employed for NT. DNMT transcripts were significantly decreased, whereas Mash2 transcripts were significantly increased in both NT groups compared to their IVP counterparts. The amount of IF mRNA was significantly higher in P8 derived than in P5/6 and IVP embryos. In experiment 4, the RA of DNMT transcripts was decreased in in vivo-derived blastocysts compared to those produced in vitro. Mash2 expression was increased in in vivo embryos and those IVP embryos produced in medium containing Sigma BSA. The RA of Hsp was higher in IVP embryos produced in serum containing medium than in those produced in Sigma BSA or in vivo. In vivo embryos and those produced in Life Technologies BSA had the lowest expression of IF transcripts. Expression of all other genes was not affected by variation in NT methodology or IVP culture systems throughout experiments 1-4. In conclusion, depending on steps of the cloning procedure NT-derived embryos display marked differences from their IVP- and in vivo-derived counterparts. An aberrant expression pattern in NT embryos was found with respect to genes thought to be involved in stress adaptation, trophoblastic function, and DNA methylation during preimplantation development. PMID- 11420255 TI - Spermatogenesis in Bclw-deficient mice. AB - Bclw is a death-protecting member of the Bcl2 family of apoptosis-regulating proteins. Mice that are mutant for Bclw display progressive and nearly complete testicular degeneration. We performed a morphometric evaluation of testicular histopathology in Bclw-deficient male mice between 9 days postnatal (p9) through 1 yr of age. Germ cell loss began by p22, with only few germ cells remaining beyond 7 mo of age. A complete block to elongated spermatid development at step 13 occurred during the first wave of spermatogenesis, whereas other types of germ cells were lost sporadically. Depletion of Sertoli cells commenced between p20 and p23 and continued until 1 yr of age, when few, if any, Sertoli cells remained. Mitochondria appeared to be swollen and the cytoplasm dense by electron microscopy, but degenerating Bclw-deficient Sertoli cells failed to display classical features of apoptosis, such as chromatin condensation and nuclear fragmentation. Macrophages entered seminiferous tubules and formed foreign-body giant cells that engulfed and phagocytosed the degenerated Sertoli cells. Leydig cell hyperplasia was evident between 3 and 5 mo of age. However, beginning at 7 mo of age, Leydig cells underwent apoptosis, with dead cells being phagocytosed by macrophages. The aforementioned cell losses culminated in a testis-containing vasculature, intertubular phagocytic cells, and peritubular cell "ghosts." An RNA in situ hybridization study indicates that Bclw is expressed in Sertoli cells in the adult mouse testis. Consequently, the diploid germ cell death may be an indirect effect of defective Sertoli cell function. Western analysis was used to confirm that Bclw is not expressed in spermatids; thus, loss of this cell type most likely results from defective Sertoli cell function. Because Bclw does not appear to be expressed in Leydig cells, loss of Leydig cells in Bclw-deficient mice may result from depletion of Sertoli cells. Bclw-deficient mice serve as a unique model to study homeostasis of cell populations in the testis. PMID- 11420256 TI - Why is apoptosis important to clinicians? PMID- 11420257 TI - Depression, suicide, and the national service framework. PMID- 11420258 TI - Environmental factors in stroke rehabilitation. PMID- 11420259 TI - Commission for Health Improvement invents itself. PMID- 11420260 TI - Racism in medicine. PMID- 11420261 TI - Scientists screen embryo for genetic predisposition to cancer. PMID- 11420262 TI - Ethnic minority doctors hit glass ceiling in NHS. PMID- 11420264 TI - Compromise plan put forward for GMC. PMID- 11420265 TI - US judge condemns health plan for not covering contraception. PMID- 11420271 TI - Multidisciplinary rehabilitation for chronic low back pain: systematic review. AB - OBJECTIVE: To assess the effect of multidisciplinary biopsychosocial rehabilitation on clinically relevant outcomes in patients with chronic low back pain. DESIGN: Systematic literature review of randomised controlled trials. PARTICIPANTS: A total of 1964 patients with disabling low back pain for more than three months. MAIN OUTCOME MEASURES: Pain, function, employment, quality of life, and global assessments. RESULTS: Ten trials reported on a total of 12 randomised comparisons of multidisciplinary treatment and a control condition. There was strong evidence that intensive multidisciplinary biopsychosocial rehabilitation with functional restoration improves function when compared with inpatient or outpatient non-multidisciplinary treatments. There was moderate evidence that intensive multidisciplinary biopsychosocial rehabilitation with functional restoration reduces pain when compared with outpatient non-multidisciplinary rehabilitation or usual care. There was contradictory evidence regarding vocational outcomes of intensive multidisciplinary biopsychosocial intervention. Some trials reported improvements in work readiness, but others showed no significant reduction in sickness leaves. Less intensive outpatient psychophysical treatments did not improve pain, function, or vocational outcomes when compared with non-multidisciplinary outpatient therapy or usual care. Few trials reported effects on quality of life or global assessments. CONCLUSIONS: The reviewed trials provide evidence that intensive multidisciplinary biopsychosocial rehabilitation with functional restoration reduces pain and improves function in patients with chronic low back pain. Less intensive interventions did not show improvements in clinically relevant outcomes. PMID- 11420272 TI - Population based intervention to change back pain beliefs and disability: three part evaluation. AB - OBJECTIVE: To evaluate the effectiveness of a population based, state-wide public health intervention designed to alter beliefs about back pain, influence medical management, and reduce disability and costs of compensation. DESIGN: Quasi experimental, non-randomised, non-equivalent, before and after telephone surveys of the general population and postal surveys of general practitioners with an adjacent state as control group and descriptive analysis of claims database. SETTING: Two states in Australia. PARTICIPANTS: 4730 members of general population before and two and two and a half years after campaign started, in a ratio of 2:1:1; 2556 general practitioners before and two years after campaign onset. MAIN OUTCOME MEASURES: Back beliefs questionnaire, knowledge and attitude statements about back pain, incidence of workers' financial compensation claims for back problems, rate of days compensated, and medical payments for claims related to back pain and other claims. RESULTS: In the intervention state beliefs about back pain became more positive between successive surveys (mean improvement in questionnaire score 1.9 (95% confidence interval 1.3 to 2.5), P<0.001 and 3.2 (2.6 to 3.9), P<0.001, between baseline and the second and third survey, respectively). Beliefs about back pain also improved among doctors. There was a clear decline in number of claims for back pain, rates of days compensated, and medical payments for claims for back pain over the duration of the campaign. CONCLUSIONS: A population based strategy of provision of positive messages about back pain improves population and general practitioner beliefs about back pain and seems to influence medical management and reduce disability and workers' compensation costs related to back pain. PMID- 11420273 TI - Questionnaire survey of urinary incontinence in women with cystic fibrosis. AB - Cystic fibrosis is an inherited disease characterised by the production of infected secretions, and it requires lifelong daily treatment by airway clearance. We knew that some women attending our clinic for adults with cystic fibrosis leaked urine when performing airway clearance or spirometry, but they were dismissive during discussion and were rarely forthcoming about the problem. This study was designed to determine the prevalence of urinary incontinence in women with cystic fibrosis, to establish the importance of the problem as perceived by the patients, and to identify those women who wanted help. PMID- 11420274 TI - Modernising primary and community health services. PMID- 11420275 TI - The role of apoptosis in autoimmune thyroid disorders and thyroid cancer. PMID- 11420276 TI - Studies of apoptosis in breast cancer. PMID- 11420278 TI - HIV counselling and the psychosocial management of patients with HIV or AIDS. PMID- 11420279 TI - What is apoptosis, and why is it important? PMID- 11420280 TI - How apoptosis is regulated, and what goes wrong in cancer. PMID- 11420281 TI - Does apoptosis have a role in neurodegeneration? PMID- 11420294 TI - Calcineurin, mitochondrial membrane potential, and cardiomyocyte apoptosis. PMID- 11420295 TI - Ref-1 and transcriptional control of endothelial apoptosis. PMID- 11420296 TI - Microarrays : managing the data deluge. PMID- 11420297 TI - Hypoxic pulmonary vasoconstriction: a radical view. PMID- 11420298 TI - Developing a strategy to define the effects of insulin-like growth factor-1 on gene expression profile in cardiomyocytes. AB - Insulin-like growth factor (IGF)-1 activates intracellular signaling pathways and regulates myocardial structure and function. This study used DNA microarray to define the effects of IGF-1 on gene expression in cardiomyocytes. Despite DNA microarray becoming a popular tool for profiling gene expression, the specificity of DNA microarray results is rarely addressed. Our data showed that the specificity of a DNA microarray study can be increased by repetitive experiments and by excluding minimally expressed genes. In this study, the false-positive rates were reduced to <0.2%. Future DNA microarray studies should incorporate a proper strategy to minimize false-positive results. IGF-1 modulates the expression of genes in 17 functional categories, but most genes clustered around the regulation of intracellular signaling, cell cycle, transcription/translation, cellular respiration and mitochondrial function, cell survival, ion channels and calcium signaling, and humoral factors. To further explore whether extracellular signal-regulated kinase (ERK) and phosphatidylinositol (PI) 3 kinase specifically regulate different sets of genes, the effects of IGF-1 were inhibited with PD98059 or LY294002. The results showed that the majority of genes regulated by IGF-1 required activation of both ERK and PI 3 kinase. Thus, PI 3 kinase and ERK coordinately mediate the transcriptional regulatory effects of IGF-1 in cardiac muscle cells. These findings provide novel insight into how IGF-1 signaling modulates the programming of cardiac muscle gene expression. PMID- 11420299 TI - Calcineurin pathway is required for endothelin-1-mediated protection against oxidant stress-induced apoptosis in cardiac myocytes. AB - Endothelin-1 (ET-1) acts not only as a growth-promoting peptide but also as a potent survival factor against myocardial cell apoptosis. However, the signaling pathways leading to myocardial cell protection by ET-1 are poorly understood. Using a culture system of primary cardiac myocytes derived from neonatal rats, we show in the present study that ET-1 almost completely blocked the hydrogen peroxide-induced increase in the percentage of TdT-mediated dUTP-biotin nick-end labeling-positive myocytes. Apoptosis inhibition by ET-1 was confirmed by cytofluorometric analysis as well as by examination of the ladder formation, morphological features, and caspase-3 cleavage. We have found that ET-1 converts the nuclear factor of activated T lymphocytes (NFATc) in cardiac myocytes into high-mobility forms and translocates cytoplasmic NFATc to the nuclei. In addition, ET-1 stimulates the interaction between NFATc and the cardiac restricted zinc-finger protein GATA4 in these cells. The immunosuppressants cyclosporin A and FK506, which antagonize calcineurin, negated the inhibitory effect of ET-1 on apoptosis. Calcineurin activation de novo was sufficient to inhibit hydrogen peroxide-induced apoptosis. ET-1 induced the expression of an antiapoptotic protein bcl-2 in cardiac myocytes in a cyclosporin A-dependent manner, but it did not alter the expression of bax. Cyclosporin A also attenuated the ET-1-stimulated transcription of the bcl-2 gene in these cells. These findings demonstrate that the calcineurin pathway is required for the inhibitory effect of ET-1 on oxidant stress-induced apoptosis in cardiac myocytes. PMID- 11420300 TI - Overexpression of Ref-1 inhibits hypoxia and tumor necrosis factor-induced endothelial cell apoptosis through nuclear factor-kappab-independent and dependent pathways. AB - We hypothesized that a redox-sensitive transcription factor, redox factor-1 (Ref 1) (HAP1, APE, and APEX), was critical in the regulation of endothelial cell survival in response to hypoxia and cytokines, including tumor necrosis factor (TNF)-alpha. Hypoxia resulted in a significant decrease in Ref-1 protein expression in both human umbilical vein endothelial cells and calf pulmonary artery endothelial cells. The hypoxia-induced decrease in Ref-1 expression was followed by a significant induction of apoptosis as measured by caspase 3 activity and nuclear morphology. Transient upregulation of Ref-1 significantly inhibited hypoxia-induced apoptosis. However, deletion of the redox-sensitive domain of Ref-1 abolished the antiapoptotic effect. We postulated that the antiapoptotic effects of Ref-1 were mediated through nuclear factor-kappaB (NF kappaB). However, blockade of NF-kappaB with a dominant-negative IkappaB (S32A/S36A) expression vector had no effect on Ref-1-mediated survival under hypoxic conditions. The second aim of this study was to test the cytoprotective ability of Ref-1 upregulation in response to TNF-induced apoptosis. Ref-1 inhibition of TNF-induced death was associated with a significant potentiation of NF-kappaB activity. Deletion of the redox-sensitive domain of Ref-1 significantly inhibited TNF-induced NF-kappaB activation. Moreover, loss of the redox-sensitive domain also abolished the antiapoptotic effect of Ref-1 in response to TNF. To test whether Ref-1 induced activation of NF-kappaB was necessary to promote survival, we blocked NF-kappaB activity with a dominant-negative IkappaB (S32A/S36A). Indeed, blockade of NF-kappaB activity abolished the ability of Ref 1 to rescue TNF-induced apoptosis. In conclusion, upregulation of Ref-1 promotes endothelial cell survival in response to hypoxia and TNF through NF-kappaB independent and NF-kappaB-dependent signaling cascades, respectively. Moreover, it seems that Ref-1 may act as a critical cofactor, mediating the TNF-induced NF kappaB response in the vascular endothelium. PMID- 11420301 TI - Sinoatrial nodal cell ryanodine receptor and Na(+)-Ca(2+) exchanger: molecular partners in pacemaker regulation. AB - The rate of spontaneous diastolic depolarization (DD) of sinoatrial nodal cells (SANCs) that triggers recurrent action potentials (APs) is a fundamental aspect of the heart's pacemaker. Here, in experiments on isolated SANCs, using confocal microscopy combined with a patch clamp technique, we show that ryanodine receptor Ca(2+) release during the DD produces a localized subsarcolemmal Ca(2+) increase that spreads in a wavelike manner by Ca(2+)-induced Ca(2+) release and produces an inward current via the Na(+)-Ca(2+) exchanger (NCX). Ryanodine, a blocker of the sarcoplasmic reticulum Ca(2+) release channel, in a dose-dependent manner reduces the SANC beating rate with an IC(50) of 2.6 micromol/L and abolishes the local Ca(2+) transients that precede the AP upstroke. In voltage-clamped cells in which the DD was simulated by voltage ramp, 3 micromol/L ryanodine decreased an inward current during the voltage ramp by 1.6+/-0.3 pA/pF (SEM, n=4) leaving the peak of L-type Ca(2+) current unchanged. Likewise, acute blockade of the NCX (via rapid substitution of bath Na(+) by Li(+)) abolished SANC beating and reduced the inward current to a similar extent (1.7+/-0.4 pA/pF, n=4), as did ryanodine. Thus, in addition to activation/inactivation of multiple ion channels, Ca(2+) activation of the NCX, because of localized sarcoplasmic reticulum Ca(2+) release, is a critical element in a chain of molecular interactions that permits the heartbeat to occur and determines its beating rate. PMID- 11420302 TI - Model for hypoxic pulmonary vasoconstriction involving mitochondrial oxygen sensing. AB - We tested whether mitochondria function as the O(2) sensor underlying hypoxic pulmonary vasoconstriction (HPV). In buffer-perfused rat lungs, rotenone, myxothiazol, and diphenyleneiodonium, which inhibit mitochondria in the proximal region of the electron transport chain (ETC), abolished HPV without attenuating the response to U46619. Cyanide and antimycin A inhibit electron transfer in the distal region of the ETC, but they did not abolish HPV. Cultured pulmonary artery (PA) myocytes contract in response to hypoxia or to U46619. The hypoxic response was abolished while the response to U46619 was maintained in mutant (rho(0)) PA myocytes lacking a mitochondrial ETC. To test whether reactive oxygen species (ROS) derived from mitochondria act as signaling agents in HPV, the antioxidants pyrrolidinedithiocarbamate and ebselen and the Cu,Zn superoxide dismutase inhibitor diethyldithiocarbamate were used. These abolished HPV without affecting contraction to U46619, suggesting that ROS act as second messengers. In cultured PA myocytes, oxidation of intracellular 2',7'-dichlorofluorescin diacetate (DCFH) dye increased under 2% O(2), indicating that myocytes increase their generation of H(2)O(2) during hypoxia. This was attenuated by myxothiazol, implicating mitochondria as the source of increased ROS during HPV. These results indicate that mitochondrial ATP is not required for HPV, that mitochondria function as O(2) sensors during hypoxia, and that ROS generated in the proximal region of the ETC act as second messengers in the response. PMID- 11420303 TI - Mitochondrial ATP-sensitive potassium channels inhibit apoptosis induced by oxidative stress in cardiac cells. AB - Mitochondria can either enhance or suppress cell death. Cytochrome c release from mitochondria and depolarization of the mitochondrial membrane potential (DeltaPsi) are crucial events in triggering apoptosis. In contrast, activation of mitochondrial ATP-sensitive potassium (mitoK(ATP)) channels prevents lethal ischemic injury in vivo, implicating these channels as key players in the process of ischemic preconditioning. We probed the relationship between mitoK(ATP) channels and apoptosis in cultured neonatal rat cardiac ventricular myocytes. Incubation with 200 micromol/L hydrogen peroxide induced TUNEL positivity, cytochrome c translocation, caspase-3 activation, poly(ADP-ribose) polymerase cleavage, and dissipation of DeltaPsi. Pharmacological opening of mitoK(ATP) channels by diazoxide (100 micromol/L) preserved mitochondrial integrity and suppressed all markers of apoptosis. Diazoxide prevented DeltaPsi depolarization in a concentration-dependent manner (EC(50) approximately 40 micromol/L, with saturation by 100 micromol/L), as shown by both flow cytometry and quantitative image analysis of cells stained with fluorescent DeltaPsi indicators. These cytoprotective effects of diazoxide were reproduced by pinacidil, another mitoK(ATP) agonist, and blocked by the mitoK(ATP) channel antagonist 5 hydroxydecanoate (500 micromol/L). Our findings identify a novel mitochondrial pathway that is protective against apoptosis. The results also pinpoint mitoK(ATP) channels as logical therapeutic targets in diseases of enhanced apoptosis and oxidative stress. PMID- 11420304 TI - Acute regulation of fatty acid oxidation and amp-activated protein kinase in human umbilical vein endothelial cells. AB - It is generally accepted that endothelial cells generate most of their ATP by anaerobic glycolysis and that very little ATP is derived from the oxidation of fatty acids or glucose. Previously, we have reported that, in cultured human umbilical vein endothelial cells (HUVECs), activation of AMP-activated protein kinase (AMPK) by the cell-permeable activator 5-aminoimidazole-4-carboximide riboside (AICAR) is associated with an increase in the oxidation of (3)H palmitate. In the present study, experiments carried out with cultured HUVECs revealed the following: (1) AICAR-induced increases in palmitate oxidation during a 2-hour incubation are associated with a decrease in the concentration of malonyl coenzyme A (CoA) (an inhibitor of carnitine palmitoyl transferase 1), which temporally parallels the increase in AMPK activity and a decrease in the activity of acetyl CoA carboxylase (ACC). (2) AICAR does not stimulate either palmitate oxidation when carnitine is omitted from the medium or oxidation of the medium-chain fatty acid octanoate. (3) When intracellular lipid pools are prelabeled with (3)H-palmitate, the measured rate of palmitate oxidation is 3 fold higher, and in the presence of AICAR, it accounts for nearly 40% of calculated ATP generation. (4) Incubation of HUVECs in a glucose-free medium for 2 hours causes the same changes in AMPK, ACC, malonyl CoA, and palmitate oxidation as does AICAR. (5) Under all conditions studied, the contribution of glucose oxidation to ATP production is minimal. The results indicate that the AMPK-ACC-malonyl CoA-carnitine palmitoyl transferase 1 mechanism plays a key role in the physiological regulation of fatty acid oxidation in HUVECs. They also indicate that HUVECs oxidize fatty acids from both intracellular and extracellular sources, and that when this is taken into account, fatty acids can be a major substrate for ATP generation. Finally, they suggest that AMPK is likely to be a major factor in modulating the response of the endothelium to stresses that alter its energy state. PMID- 11420305 TI - Distinct pathways of Ca(2+) sensitization in porcine coronary artery: effects of Rho-related kinase and protein kinase C inhibition on force and intracellular Ca(2+). AB - Alterations of the Ca(2+) sensitivity of contraction have been reported for porcine coronary artery, but the mechanisms are not clearly understood. We investigated the mechanism(s) of Ca(2+) sensitization in response to the thromboxane A(2) analogue (U46619). Our hypothesis is that different mechanisms of Ca(2+) sensitization could be distinguished by their distinct time courses. Therefore, we measured the time course of [Ca(2+)](i) and isometric force simultaneously in an intact artery after a single addition of U46619. The initial transient phase was associated with Ca(2+) release from the sarcoplasmic reticulum, whereas the maintained phase was associated with Ca(2+) influx. Two distinct types of Ca(2+) sensitization characterized these phases with either protein kinase C (PKC)-mediated or Rho-kinase-mediated mechanisms. Their effects were quite distinct on the basis of the time courses over which the sensitization was effective. PKC inhibition (1 micromol/L calphostin C) had a much greater effect in the initial phase, diminishing the size of the transient and prolonging the rise in force and the decline in [Ca(2+)](i). There were limited effects on the sustained force. Rho-kinase inhibition (10 micromol/L Y27632), in contrast, nearly abolished the sustained force but had a lesser effect on the transient phase. Neither inhibitor had any effect on the force versus [Ca(2+)](i) relations for KCl contractures. Our evidence suggests that both PKC-mediated and Rho-kinase mediated Ca(2+) sensitizations are present in coronary arteries, but the latter is dominant in thromboxane A(2) receptor-mediated contraction. PMID- 11420306 TI - Diabetes mellitus enhances vascular matrix metalloproteinase activity: role of oxidative stress. AB - Diabetes mellitus (DM) is a primary risk factor for cardiovascular disease. Although recent studies have demonstrated an important role for extracellular matrix metalloproteinases (MMPs) in atherosclerosis, little is known about the effects of hyperglycemia on MMP regulation in vascular cells. Gelatin zymography and Western blot analysis revealed that the activity and expression of 92-kDa (MMP-9) gelatinase, but not of 72 kDa (MMP-2) gelatinase, were significantly increased in vascular tissue and plasma of two distinct rodent models of DM. Bovine aortic endothelial cells (BAECs) grown in culture did not express MMP-9 constitutively; however, chronic (2-week) incubation with high glucose medium induced MMP-9 promoter activity, mRNA and protein expression, and gelatinase activity in BAECs. On the other hand, high glucose culture did not change MMP-9 activity from vascular smooth muscle cells or macrophages. Electron paramagnetic resonance studies indicate that BAECs chronically grown in high glucose conditions produce 70% more ROS than do control cells. Enhanced MMP-9 activity was significantly reduced by treatment with the antioxidants polyethylene glycol superoxide dismutase and N-acetyl-L-cysteine but not by inhibitors of protein kinase C. In conclusion, vascular MMP-9 activity is increased in DM, in part because of enhanced elaboration from vascular endothelial cells, and oxidative stress plays an important role. This novel mechanism of redox-sensitive MMP-9 expression by hyperglycemia may provide a rationale for antioxidant therapy to modulate diabetic vascular complications. PMID- 11420307 TI - Contraction and intracellular Ca(2+) handling in isolated skeletal muscle of rats with congestive heart failure. AB - A decreased exercise tolerance is a common symptom in patients with congestive heart failure (CHF). This decrease has been suggested to be partly due to altered skeletal muscle function. Therefore, we have studied contractile function and cytoplasmic free Ca(2+) concentration ([Ca(2+)](i), measured with the fluorescent dye indo 1) in isolated muscles from rats in which CHF was induced by ligation of the left coronary artery. The results show no major changes of the contractile function and [Ca(2+)](i) handling in unfatigued intact fast-twitch fibers isolated from flexor digitorum brevis muscles of CHF rats, but these fibers were markedly more susceptible to damage during microdissection. Furthermore, CHF fibers displayed a marked increase of baseline [Ca(2+)](i) during fatigue. Isolated slow-twitch soleus muscles of CHF rats displayed slower twitch contraction and tetanic relaxation than did muscles from sham-operated rats; the slowing of relaxation became more pronounced during fatigue in CHF muscles. Immunoblot analyses of sarcoplasmic reticulum proteins and sarcolemma Na(+),K(+) ATPase showed no difference in flexor digitorum brevis muscles of sham-operated versus CHF rats. In conclusion, functional impairments can be observed in limb muscle isolated from rats with CHF. These impairments seem to mainly involve structures surrounding the muscle cells and sarcoplasmic reticulum Ca(2+) pumps, the dysfunction of which becomes obvious during fatigue. PMID- 11420308 TI - Protein kinase C epsilon-Src modules direct signal transduction in nitric oxide induced cardioprotection: complex formation as a means for cardioprotective signaling. AB - An essential role for protein kinase C epsilon (PKCepsilon) has been shown in multiple forms of cardioprotection; however, there is a distinct paucity of information concerning the signaling architecture that is responsible for the manifestation of a protective phenotype. We and others have recently shown that signal transduction may proceed via the formation of signaling complexes (Circ Res. 2001;88:59-62). In order to understand if the assembly of multiprotein complexes is the manner by which signaling is conducted in cardioprotection, we designed a series of experiments to characterize the associations of Src tyrosine kinase with PKCepsilon in a conscious rabbit model of nitric oxide (NO)-induced late preconditioning. Our data demonstrate that PKCepsilon and Src can form functional signaling modules in vitro: PKCepsilon interacts with Src; the association with PKCepsilon activates Src; and adult cardiac cells receiving recombinant adenoviruses encoding PKCepsilon exhibit increased Src activity. Furthermore, our results show that NO-induced late preconditioning involved PKCepsilon-Src module formation and enhanced the enzymatic activity of PKCepsilon associated Src. Inhibition of PKC blocked cardioprotection, module formation, and PKCepsilon-associated Src activity, providing direct evidence for a functional role of the PKCepsilon-Src module in the orchestration of NO-induced cardioprotection in conscious rabbits. PMID- 11420309 TI - Quinoxalines versus indolinones as inhibitors of restenosis. PMID- 11420310 TI - Novel mechanism for Brugada syndrome: defective surface localization of an SCN5A mutant (R1432G). AB - The SCN5A gene encodes the alpha subunit of the human heart sodium channel (hH1), which plays a critical role in cardiac excitability. Mutations of SCN5A underlie Brugada syndrome, an inherited disorder that leads to ventricular fibrillation and sudden death. This study describes changes in cellular localization and functional expression of hH1 in a naturally occurring SCN5A mutation (R1432G) reported for Brugada syndrome. Using patch-clamp experiments, we show that there is an abolition of functional hH1 expression in R1432G mutants expressed in human tsA201 cells but not in Xenopus oocytes. In tsA201 cells, a conservative positively charged mutant, R1432K, produced sodium currents with normal gating properties, whereas other mutations at this site abolished functional sodium channel expression. Immunofluorescent staining and confocal microscopy showed that the wild-type alpha subunit expressed in tsA201 cells was localized to the cell surface, whereas the R1432G mutant was colocalized with calnexin within the endoplasmic reticulum. The beta(1) subunit was also localized to the cell surface in the presence of the alpha subunit; however, in its absence, the beta(1) subunit was restricted to a perinuclear localization. These results demonstrate that the disruption of SCN5A cell-surface localization is one mechanism that can account for the loss of functional sodium channels in Brugada syndrome. The full text of this article is available at http://www.circresaha.org. PMID- 11420311 TI - MinK-related peptide 1: A beta subunit for the HCN ion channel subunit family enhances expression and speeds activation. AB - The HCN family of ion channel subunits underlies the currents I(f) in heart and I(h) and I(q) in the nervous system. In the present study, we demonstrate that minK-related peptide 1 (MiRP1) is a beta subunit for the HCN family. As such, it enhances protein and current expression as well as accelerating the kinetics of activation. Because MiRP1 also functions as a beta subunit for the cardiac delayed rectifier I(Kr), these results suggest that this peptide may have the unique role of regulating both the inward and outward channels that underlie cardiac pacemaker activity. The full text of this article is available at http://www.circresaha.org. PMID- 11420312 TI - Parents and end of life decisions in neonatal practice. PMID- 11420313 TI - Consent to autopsy for neonates. AB - OBJECTIVES: To determine parents' views on autopsy after treatment withdrawal. DESIGN: Face to face interviews with 59 sets of bereaved parents (108 individual parents) for whose 62 babies there had been discussion of treatment withdrawal. RESULTS: All except one couple were asked for permission for postmortem examination; 38% refused. The main reasons for declining were concerns about disfigurement, a wish to have the child left in peace, and a feeling that an autopsy was unnecessary because the parents had no unanswered questions. The diagnosis, the age of the child, and the approach of the consultant appeared to influence consent rates. Of those who agreed to autopsies, 92% were given the results by the neonatologist concerned. Whether or not they had agreed to the procedure, at 13 months no parent expressed regrets about their decision. CONCLUSIONS: Autopsy rates in the East of Scotland stand at 62%. Parents' perceptions are an important element in consent to postmortem examination. PMID- 11420314 TI - Lingering death after treatment withdrawal in the neonatal intensive care unit. AB - OBJECTIVE: To explore parents' perceptions of treatment withdrawal and the dying process. DESIGN: Face to face interviews with 59 sets of parents of 62 babies in the East of Scotland three months and 13 months after death. RESULTS: 22% of the parents expressed reservations about the length of the dying process, which they reported in these instances had taken from three to 36 hours. Deaths that medical teams had predicted would be quick had, according to the parents' recollections, taken from 1.5 to 31 hours. When a baby died swiftly, this seemed to confirm the wisdom of the decision to stop. When babies lingered, doubts were raised. CONCLUSIONS: Parents need to be adequately prepared for what may happen after treatment withdrawal. The debate should be reopened about the best way to manage protracted deaths in line with parental need. PMID- 11420315 TI - Effect of maternal tocolysis on the incidence of severe periventricular/intraventricular haemorrhage in very low birthweight infants. AB - AIM: To examine the relation between grade III-IV periventricular/intraventricular haemorrhage (PVH/IVH) and antenatal exposure to tocolytic treatment in very low birthweight (VLBW) premature infants. STUDY DESIGN: The study population consisted of 2794 infants from the Israel National VLBW Infant Database, of gestational age 24-32 weeks, who had a cranial ultrasound examination during the first 28 days of life. Infants of mothers with pregnancy induced hypertension or those exposed to more than one tocolytic drug were excluded. Of the 2794 infants, 2013 (72%) had not been exposed to tocolysis and 781 (28%) had been exposed to a single tocolytic agent. To evaluate the effect of tocolysis and confounding variables on grade III-IV PVH/IVH, the chi(2) test, univariate analysis, and a logistic regression model were used. RESULTS: Of the 781 infants (28%) exposed to tocolysis, 341 (12.2%) were exposed to magnesium sulphate, 263 (9.4%) to ritodrine, and 177 (6.3%) to indomethacin. The overall incidence of grade III-IV PVH/IVH was 13.4%. In the multivariate logistic regression analysis, the following factors were related significantly and independently to grade III-IV PVH/IVH: no prenatal steroid treatment, low gestational age, one minute Apgar score 0-3, respiratory distress syndrome, patent ductus arteriosus, mechanical ventilation, and pneumothorax. Infants exposed to ritodrine tocolysis (but not to the other tocolytic drugs) were at significantly lower risk of grade III-IV PVH/IVH after adjustment for other variables (odds ratio = 0.3; 95% confidence interval 0.2 to 0.6). CONCLUSION: This study suggests that antenatal exposure of VLBW infants to ritodrine tocolysis, in contrast with tocolysis induced by magnesium sulphate or indomethacin, was associated with a lower incidence of grade III-IV PVH/IVH. PMID- 11420316 TI - Treatment of neonatal thrombus formation with recombinant tissue plasminogen activator: six years experience and review of the literature. AB - BACKGROUND: Thrombosis is a relatively rare event in children. However, many conditions in the neonatal period result in an increased risk of thrombus formation. The major risk factor is the indwelling intravascular catheter. Numerous small studies have reported experience of thrombolytic treatment for neonatal thrombotic disease with a wide range of different thrombolytic agents in various forms of administration, dosage, and duration, but no conclusions on the most effective treatment for neonates has been reached. OBJECTIVE: To assess the efficacy and safety of thrombolytic treatment of neonatal catheter related thrombus (CRT) formation with recombinant tissue plasminogen activator (rt-PA). METHOD: Over a six year period, 14 neonates with CRT were treated with the same rt-PA protocol (an initial bolus of 0.7 mg/kg over 30-60 minutes followed by infusion of 0.2 mg/kg/h). RESULTS: Complete clot dissolution was documented in 11 patients, and partial clot lysis in two patients, leading to a patency rate of 94%. In two cases, local bleeding occurred, resulting in treatment failure in one case. Finally, antithrombin III substitution was required in one case. No other complications such as severe bleeding were recognised. CONCLUSION: With the use of close clinical and haematological monitoring on a neonatal intensive care unit combined with serial two dimensional colour echocardiography, the present rt-PA protocol was shown to be a safe and effective method of clot dissolution in neonates. PMID- 11420317 TI - Educational and behavioural problems in babies of 32-35 weeks gestation. AB - AIM: To identify incidence of school and behaviour problems at age 7 years in children born between 32 and 35 weeks gestation, and investigate perinatal risk factors. METHOD: The study population consisted of all children born at 32-35 weeks gestation to mothers resident in Oxfordshire in 1990. General practitioners, parents, and teachers were asked about health, behaviour, and education by postal questionnaire. Teachers rated children on level of function in six areas using a five point scale. They also completed the Strengths and Difficulties behaviour questionnaire. Perinatal risk factors were identified for children with poor school performance using a univariate and multivariate analysis. RESULTS: Teacher responses were obtained for 117 (66%) of the 176 children in the cohort. Twenty nine (25%) required support from a non-teaching assistant, five (4%) had required a statement of special educational needs, and three (3%) were at special school. Poor outcome was reported for 32% in writing, 31% in fine motor skills, 29% in mathematics, 19% in speaking, 21% in reading, and 12% in physical education. On the behaviour questionnaire, 19% of the cohort achieved an abnormal hyperactivity score (population norm 10%). Multivariate analysis showed perinatal variables that remained significant, independent of other variables; they were discharge from the special care baby unit > 36 weeks postconceptional age (odds ratio 4.15; 95% confidence interval 1.43 to 12.05) and male sex (odds ratio 3.88; 95% confidence interval 1.42 to 10.6). CONCLUSION: Up to a third of children born between 32 and 35 weeks gestation may have school problems. As there are larger numbers in this gestational category compared with smaller babies, this finding has implications for educational services. PMID- 11420318 TI - Randomised controlled trial of postnatal sodium supplementation in infants of 25 30 weeks gestational age: effects on cardiopulmonary adaptation. AB - BACKGROUND: It has previously been shown that, in preterm babies, routine sodium supplementation from 24 hours after birth is associated with increased risk of oxygen dependency and persistent expansion of the extracellular compartment. OBJECTIVE: To explore whether this is mediated by a delayed fall in pulmonary artery pressure (PAP). Postnatal changes in PAP, estimated as the ratio of time to peak velocity to right ventricular ejection time, corrected for heart rate (TPV:RVET(c)), were compared in preterm infants who received routine sodium supplements that were either early or delayed. METHODS: Infants were randomised, stratified according to sex and gestation, to receive a sodium intake of 4 mmol/kg/day starting either from 24 hours after birth or when a weight loss of 6% of birth weight was achieved. Echocardiographic assessment was made on the day of delivery (day 0), and on days 1, 2, 7, and 14. Babies with congenital heart disease were excluded. RESULTS: There was no difference between the two groups in TPV:RVET(c) measured sequentially after birth. On within group testing, when compared with values at birth, the ratio was higher by day 3 in the early supplemented group, suggesting a more rapid fall in PAP compared with the late supplemented group, in whom a significant fall did not occur until day 14. CONCLUSIONS: The timing of sodium supplementation after preterm birth does not appear to affect the rate of fall in PAP as measured by the TPV:RVET(c) ratio. The previous observation linking routine sodium supplementation from 24 hours after birth with increased risk of continuing oxygen requirement therefore does not appear to be mediated by a delayed fall in PAP. Instead, the increased risk of continuing oxygen requirement is likely to be a direct consequence of persistent expansion of the extracellular compartment and increased pulmonary interstitial fluid, resulting from a sodium intake that exceeded sodium excretory capacity. This adds further weight to the view that clinical management, in this case the timing of routine sodium supplementation, should be individually tailored and delayed until the onset of postnatal extracellular volume contraction, marked clinically by weight loss. PMID- 11420319 TI - Factors affecting the incidence of chronic lung disease of prematurity in 1987, 1992, and 1997. AB - OBJECTIVE: To determine changes in the incidence of chronic lung disease of prematurity between 1987, 1992, and 1997. METHODS: Observational study based on data derived from a geographically defined population: Trent Health Region, United Kingdom. Three time periods were compared: 1 February 1987 to 31 January 1988 (referred to as 1987); 1 April 1992 to 31 March 1993 (referred to as 1992); 1997. All infants of < or = 32 completed weeks gestation born to Trent resident mothers within the study periods and admitted to a neonatal unit were included. Rates of chronic lung disease were determined using two definitions: (a) infants who remained dependent on active respiratory support or increased oxygen at 28 days of age; (b) infants who remained dependent on active respiratory support or increased oxygen at a corrected age of 36 weeks gestation. RESULTS: Between 1987 and 1992 there was a fall in the birth rate, but a significant increase was noted in the number of babies of < or = 32 weeks gestation admitted to a neonatal unit. There was no significant change in survival when the two groups of infants were directly compared. However, mean gestation and birth weight fell. Adjusting for this change showed a significant improvement in survival (28 day survival: odds ratio (OR) = 1.69; 95% confidence interval (95% CI) = 1.23 to 2.33. Survival to 36 week corrected gestation: OR = 1.45; 95% CI = 1.06 to 1.98). These changes were accompanied by a large increase in the incidence of chronic lung disease even after allowing for the change in population characteristics (28 day definition: OR = 2.20; 95% CI = 1.47 to 3.30. 36 week definition: OR = 3.04; 95% CI = 1.91 to 4.83). Between 1992 and 1997 a different pattern emerged. There was a further increase in the number of babies admitted for neonatal care at 0.95 for all models. LM, on the other hand, reconstructs the correct bifurcating tree with a probability of >0.95 for most branch length combinations even under models with varying substitution rates. The parameter domain for which GM recovers the true tree is much smaller. When the exterior branch lengths are larger than a (analytically derived) threshold value depending on the tree shape (rather than the evolutionary model), GM reconstructs a star tree rather than the true tree. We suggest a combined approach of LM and GM for the evaluation of starlike trees. This approach offers the possibility of testing for significant positive interior branch lengths without extensive statistical and computational efforts. PMID- 11420362 TI - Molecular insights into the evolution of the family Bovidae: a nuclear DNA perspective. AB - The evolutionary history of the family Bovidae remains controversial despite past comprehensive morphological and genetic investigations. In an effort to resolve some of the systematic uncertainties within the group, a combined molecular phylogeny was constructed based on four independent nuclear DNA markers (2,573 characters) and three mitochondrial DNA genes (1,690 characters) for 34 bovid taxa representing all seven of the currently recognized bovid subfamilies. The nuclear DNA fragments were analyzed separately and in combination after partition homogeneity tests were performed. There was no significant rate heterogeneity among lineages, and retention index values indicated the general absence of homoplasy in the nuclear DNA data. The conservative nuclear DNA data were remarkably effective in resolving associations among bovid subfamilies, which had a rapid radiation dating back to approximately 23 MYA. All analyses supported the monophyly of the Bovinae (cow, nilgai, and kudu clade) as a sister lineage to the remaining bovid subfamilies, and the data convincingly suggest that the subfamilies Alcelaphinae (hartebeest, tsessebe, and wildebeest group) and Hippotraginae (roan, sable, and gemsbok clade) share a close evolutionary relationship and together form a sister clade to the more primitive Caprinae (represented by sheep, goat, and muskox). The problematic Reduncinae (waterbuck, reedbuck) seem to be the earliest-diverging group of the Caprinae/Alcelaphinae/Hippotraginae clade, whereas the Antilopinae (gazelle and dwarf antelope clade) were always polyphyletic. The sequence data suggest that the initial diversification of the Bovidae took place in Eurasia and that lineages such as the Cephalophinae and other enigmatic taxa (impala, suni, and klipspringer) most likely originated, more or less contemporaneously, in Africa. PMID- 11420363 TI - HIV-1 and HIV-2 LTR nucleotide sequences: assessment of the alignment by N-block presentation, "retroviral signatures" of overrepeated oligonucleotides, and a probable important role of scrambled stepwise duplications/deletions in molecular evolution. AB - Previous analyses of retroviral nucleotide sequences, suggest a so-called "scrambled duplicative stepwise molecular evolution" (many sectors with successive duplications/deletions of short and longer motifs) that could have stemmed from one or several starter tandemly repeated short sequence(s). In the present report, we tested this hypothesis by focusing on the long terminal repeats (LTRs) (and flanking sequences) of 24 human and 3 simian immunodeficiency viruses. By using a calculation strategy applicable to short sequences, we found consensus overrepresented motifs (often containing CTG or CAG) that were congruent with the previously defined "retroviral signature." We also show many local repetition patterns that are significant when compared with simply shuffled sequences. First- and second-order Markov chain analyses demonstrate that a major portion of the overrepresented oligonucleotides can be predicted from the dinucleotide compositions of the sequences, but by no means can biological mechanisms be deduced from these results: some of the listed local repetitions remain significant against dinucleotide-conserving shuffled sequences; together with previous results, this suggests that interspersed and/or local mononucleotide and oligonucleotide repetitions could have biased the dinucleotide compositions of the sequences. We searched for suggestive evolutionary patterns by scrutinizing a reliable multiple alignment of the 27 sequences. A manually constructed alignment based on homology blocks was in good agreement with the polypeptide alignment in the coding sectors and has been exhaustively assessed by using a multiplied alphabet obtained by the promising mathematical strategy called the N-block presentation (taking into account the environment of each nucleotide in a sequence). Sector by sector, we hypothesize many successive duplication/deletion scenarios that fit our previous evolutionary hypotheses. This suggests an important duplication/deletion role for the reverse transcriptase, particularly in inducing stuttering cryptic simplicity patterns. PMID- 11420364 TI - Gene genealogies, cryptic species, and molecular evolution in the human pathogen Coccidioides immitis and relatives (Ascomycota, Onygenales). AB - Previous genealogical analyses of population structure in Coccidioides immitis revealed the presence of two cryptic and sexual species in this pathogenic fungus but did not clarify their origin and relationships with respect to other taxa. By combining the C. immitis data with those of two of its closest relatives, the free-living saprophytes Auxarthron zuffianum and Uncinocarpus reesii, we show that the C. immitis species complex is monophyletic, indicating a single origin of pathogenicity. Cryptic species also were found in both A. zuffianum and U. reesii, indicating that they can be found in both pathogenic and free-living fungi. Our study, together with a few others, indicates that the current list of known fungal species might be augmented by a factor of at least two. However, at least in the C. immitis, A. zuffianum, and U. reesii complexes, cryptic species represent subdivisions at the tips of deep monophyletic clades and thus well within the existing framework of generic classification. An analysis of silent and expressed divergence and polymorphism values between and within the taxa identified by genealogical concordance did not reveal faster evolution in C. immitis as a consequence of adaptation to the pathogenic habit, nor did it show positive Darwinian evolution in a region of a dioxygenase gene (tcrP gene coding for 4-HPPD) known to cause antigenic responses in humans. Instead, the data suggested relative stasis, indicative of purifying selection against mostly deleterious mutations. Two introns in the same gene fragment were considerably more divergent than exons and were unalignable between species complexes but had very low polymorphism within taxa. PMID- 11420365 TI - Y-chromosome mismatch distributions in Europe. AB - Ancient demographic events can be inferred from the distribution of pairwise sequence differences (or mismatches) among individuals. We analyzed a database of 3,677 Y chromosomes typed for 11 biallelic markers in 48 human populations from Europe and the Mediterranean area. Contrary to what is observed in the analysis of mitochondrial polymorphisms, Tajima's test was insignificant for most Y chromosome samples, and in 47 populations the mismatch distributions had multiple peaks. Taken at face value, these results would suggest either (1) that the size of the male population stayed essentially constant over time, while the female population size increased, or (2) that different selective regimes have shaped mitochondrial and Y-chromosome diversity, leading to an excess of rare alleles only in the mitochondrial genome. An alternative explanation would be that the 11 variable sites of the Y chromosome do not provide sufficient statistical power, so a comparison with mitochondrial data (where more than 200 variable sites are studied in Europe) is impossible at present. To discriminate between these possibilities, we repeatedly analyzed a European mitochondrial database, each time considering only 11 variable sites, and we estimated mismatch distributions in stable and growing populations, generated by simulating coalescent processes. Along with theoretical considerations, these tests suggest that the difference between the mismatch distributions inferred from mitochondrial and Y-chromosome data are not a statistical artifact. Therefore, the observed mismatch distributions appear to reflect different underlying demographic histories and/or selective pressures for maternally and paternally transmitted loci. PMID- 11420366 TI - Hsp70 duplication in the Drosophila melanogaster species group: how and when did two become five? AB - To determine how the modern copy number (5) of hsp70 genes in Drosophila melanogaster evolved, we localized the duplication events that created the genes in the phylogeny of the melanogaster group, examined D. melanogaster genomic sequence to investigate the mechanisms of duplication, and analyzed the hsp70 gene sequences of Drosophila orena and Drosophila mauritiana. The initial two-to four hsp70 duplication occurred 10--15 MYA, according to fixed in situ hybridization to polytene chromosomes, before the origin and divergence of the melanogaster and five other species subgroups of the melanogaster group. Analysis of more than 30 kb of flanking sequence surrounding the hsp70 gene clusters suggested that this duplication was likely a retrotransposition. For the melanogaster subgroup, Southern hybridization and an hsp70 restriction map confirmed the conserved number (4) and arrangement of hsp70 genes in the seven species other than D. melanogaster. Drosophila melanogaster is unique; tandem duplication and gene conversion at the derived cluster yielded a fifth hsp70 gene. The four D. orena hsp70 genes are highly similar and concertedly evolving. In contrast, the D. mauritiana hsp70 genes are divergent, and many alleles are nonfunctional. The proliferation, concerted evolution, and maintenance of functionality in the D. melanogaster hsp70 genes is consistent with the action of natural selection in this species. PMID- 11420367 TI - The yeast protein interaction network evolves rapidly and contains few redundant duplicate genes. AB - In this paper, the structure and evolution of the protein interaction network of the yeast Saccharomyces cerevisiae is analyzed. The network is viewed as a graph whose nodes correspond to proteins. Two proteins are connected by an edge if they interact. The network resembles a random graph in that it consists of many small subnets (groups of proteins that interact with each other but do not interact with any other protein) and one large connected subnet comprising more than half of all interacting proteins. The number of interactions per protein appears to follow a power law distribution. Within approximately 200 Myr after a duplication, the products of duplicate genes become almost equally likely to (1) have common protein interaction partners and (2) be part of the same subnetwork as two proteins chosen at random from within the network. This indicates that the persistence of redundant interaction partners is the exception rather than the rule. After gene duplication, the likelihood that an interaction gets lost exceeds 2.2 x 10(-3)/Myr. New interactions are estimated to evolve at a rate that is approximately three orders of magnitude smaller. Every 300 Myr, as many as half of all interactions may be replaced by new interactions. PMID- 11420368 TI - The complete mitochondrial DNA sequence of the basal hexapod Tetrodontophora bielanensis: evidence for heteroplasmy and tRNA translocations. AB - We present the complete 15,455-nt mitochondrial DNA sequence of the springtail Tetrodontophora bielanensis (Arthropoda, Hexapoda, Collembola). The gene content is typical of most metazoans, with 13 protein-coding genes (PCGs), 2 genes encoding for ribosomal RNA subunits, and 22 tRNA genes. The nucleotide sequence shows the well-known A+T bias typical of insect mtDNA; its A+T content is lower (72.7%) than that observed in other insect species, but still higher than that in other arthropodan taxa. The bias appears to be uniform across the whole molecule, unlike other insect taxa, which show increased A+T content in the so-called A+T rich region. However, the bias is slightly higher in the third codon positions of the PCGs (81.4%). Anomalous initiation codons have been observed in the nad2 and the cox1 genes. In the latter, the ATTTAA hexanucleotide is suggested to be involved in the initiation signaling. All tRNAs could be folded into the typical cloverleaf secondary structure, but the tRNA for cysteine appears to be missing the DHU arm. Long tandemly repeated regions (193 nt) were found in the A+T-rich region, which in turn was shown to have the possibility of forming a complex array of secondary structures. One of these structures encompassed the junction between the repeats. The A+T-rich region was also interesting in that it showed heteroplasmy in the number of repeats. Three haplotypes were found, possessing 2, 3, and 4 identical repeats, respectively. The order of protein coding and rRNA genes in the molecule was determined and was identical to that of all insects studied so far. However, two tRNA translocations were found which were unprecedented among Arthropoda. These involved the trnQ, which was found between the rrnS and the A+T-rich region, and the trnS(ucn), which was located between trnM and trnI. A preliminary phylogenetic analysis based on the amino acid sequence of the PCGs failed to find support for the monophyly of Hexapoda. PMID- 11420369 TI - Gene duplication events producing muscle (M) and brain (B) isoforms of cytoplasmic creatine kinase: cDNA and deduced amino acid sequences from two lower chordates. AB - Creatine kinase (CK) is coded for by at least four loci in higher vertebrates- two cytoplasmic isoforms, muscle (M) and brain (B), and two mitochondrial isoforms, sarcomeric and ubiquitous. M is expressed primarily in skeletal muscle, while B is expressed in a variety of cells, including cardiac and smooth muscle fibers, neurons, transport epithelia, and photoreceptors. M and B subunits form very stable homodimers (MM [M-CK], BB [B-CK]) and heterodimers (MB). M-CK is capable of binding to the M line of the myofibril, thereby creating an energy transfer microcompartment; BB and MB CKs are not. M- and B-like CKs are present in all vertebrates yet examined, including fish. Cytoplasmic, dimeric CKs are widely distributed in the invertebrates. The only available amino acid sequence for an invertebrate dimeric CK, that of the protostome polychaete Chaetopterus variopedatus, is just as similar to the vertebrate M isoform as to the B isoform. Echinoderms lack dimeric, cytoplasmic CKs, which appear to be replaced by a dimeric arginine kinase which evolved secondarily from CK. Thus, it is likely that the gene duplication event producing the M and B isoforms occurred after the divergence of the chordates from echinoderms. To narrow down the timing of this duplication event, we obtained the cDNA and deduced amino acid sequences of dimeric CKs from the tunicate Ciona intestinalis (subphylum Urochordata) and the lancelet Branchiostoma floridae (subphylum Cephalochordata). Our results show that these CKs are strikingly similar to both invertebrate and vertebrate CKs. However, phylogenetic analyses by neighbor-joining and parsimony show that these two enzymes appeared to have diverged before the point of divergence of the M and B isoforms. Thus, the gene duplication event for formation of the muscle and brain isoforms of CK most likely occurred during the radiation of the fish, a time noted for gene duplication events at a variety of other loci. PMID- 11420370 TI - The evolutionary history of the coral genus Acropora (Scleractinia, Cnidaria) based on a mitochondrial and a nuclear marker: reticulation, incomplete lineage sorting, or morphological convergence? AB - This study examines molecular relationships across a wide range of species in the mass spawning scleractinian coral genus Acropora. Molecular phylogenies were obtained for 28 species using DNA sequence analyses of two independent markers, a nuclear intron and the mtDNA putative control region. Although the compositions of the major clades in the phylogenies based on these two markers were similar, there were several important differences. This, in combination with the fact that many species were not monophyletic, suggests either that introgressive hybridization is occurring or that lineage sorting is incomplete. The molecular tree topologies bear little similarity to the results of a recent cladistic analysis based on skeletal morphology and are at odds with the fossil record. We hypothesize that these conflicting results may be due to the same morphology having evolved independently more than once in Acropora and/or the occurrence of extensive interspecific hybridization and introgression in combination with morphology being determined by a small number of genes. Our results indicate that many Acropora species belong to a species complex or syngameon and that morphology has little predictive value with regard to syngameon composition. Morphological species in the genus often do not correspond to genetically distinct evolutionary units. Instead, species that differ in timing of gamete release tend to constitute genetically distinct clades. PMID- 11420371 TI - Duplication and concerted evolution of the mitochondrial control region in the parrot genus Amazona. AB - We report a duplication and rearrangement of the mitochondrial genome involving the control region of parrots in the genus Amazona. This rearrangement results in a gene order of cytochrome b/tRNA(Thr)/pND6/pGlu/CR1/tRNA(Pro)/NADH dehydrogenase 6/tRNA(Glu)/CR2/tRNA(Phe)/12s rRNA, where CR1 and CR2 refer to duplicate control regions, and pND6 and pGlu indicate presumed pseudogenes. In contrast to previous reports of duplications involving the control regions of birds, neither copy of the parrot control region shows any indications of degeneration. Rather, both copies contain many of the conserved sequence features typically found in avian control regions, including the goose hairpin, TASs, the F, C, and D boxes, conserved sequence box 1 (CSB1), and an apparent homolog to the mammalian CSB3. We conducted a phylogenetic analysis of homologous portions of the duplicate control regions from 21 individuals representing four species of Amazona (A. ochrocephala, A. autumnalis, A. farinosa, and A. amazonica) and Pionus chalcopterus. This analysis revealed that an individual's two control region copies (i.e., the paralogous copies) were typically more closely related to one another than to corresponding segments of other individuals (i.e., the orthologous copies). The average sequence divergence of the paralogous control region copies within an individual was 1.4%, versus a mean value of 4.1% between control region orthologs representing nearest phylogenetic neighbors. No differences were found between the paralogous copies in either the rate or the pattern in which the two copies accumulated base pair changes. This pattern suggests concerted evolution of the two control regions, perhaps through occasional gene conversion events. We estimated that gene conversion events occurred on average every 34,670 +/- 18,400 years based on pairwise distances between the paralogous control region sequences of each individual. Our results add to the growing body of work indicating that under some circumstances duplicated mitochondrial control regions are retained through evolutionary time rather than degenerating and being lost, presumably due to selection for a small mitochondrial genome. PMID- 11420372 TI - The frequency distribution of nucleotide variation in Drosophila simulans. AB - Patterns of codon bias in Drosophila suggest that silent mutations can be classified into two types: unpreferred (slightly deleterious) and preferred (slightly beneficial). Results of previous analyses of polymorphism and divergence in Drosophila simulans were interpreted as supporting a mutation selection-drift model in which slightly deleterious, silent mutants make significantly greater contributions to polymorphism than to divergence. Frequencies of unpreferred polymorphisms were inferred to be lower than frequencies of other silent polymorphisms. Here, I analyzed additional D. simulans data to reevaluate the support for these ideas. I found that D. simulans has fixed more unpreferred than preferred mutations, suggesting that this lineage has not been at mutation-selection-drift equilibrium at silent sites. Frequencies of polarized unpreferred polymorphisms are not skewed toward rare alleles. However, frequencies of unpolarized unpreferred codons are lower in high-bias genes than in low-bias genes. This supports the idea that unpreferred codons are borderline deleterious mutations. Purifying selection on silent sites appears to be stronger at twofold-degenerate codons than at fourfold-degenerate codons. Finally, I found that X-linked polymorphisms occur at a higher average frequency than polymorphisms on chromosome arm 3R, even though an average X-linked site is significantly less likely to be polymorphic than an average site on 3R. This result supports a previous analysis of D. simulans indicating different population genetics of X-linked versus autosomal mutations. PMID- 11420373 TI - Evidence for recent population expansion in the evolutionary history of the malaria vectors Anopheles arabiensis and Anopheles gambiae. AB - Gene flow in malaria vectors is usually estimated based on differentiation indices (e.g., F(ST)) in order to predict the contemporary spread of genes such as those conferring resistance to insecticides. This approach is reliant on a number of assumptions, the most crucial, and the one most likely to be violated in these species, being mutation-migration-drift equilibrium. Tests of this assumption for the African malaria vectors Anopheles gambiae and Anopheles arabiensis are the focus of this study. We analyzed variation at 18 microsatellite loci and the ND5 region of the mitochondrial genome in two populations of each species. Equilibrium was rejected by six of eight tests for the A. gambiae population from western Kenya and by three tests in eastern Kenya. In western Kenya, all departures from equilibrium were consistent with a recent population expansion, but in eastern Kenya, there were traces of a recent expansion and a bottleneck. Equilibrium was also rejected by two of the eight tests for both A. arabiensis populations; the departure from equilibrium was consistent with an expansion. These multiple-locus tests detected a genomewide effect and therefore a demographic event rather than a locus-specific effect, as would be caused by selection. Disequilibrium due to a recent expansion in these species implies that rates of gene flow, as inferred from differentiation indices, are overestimates as they include a historical component. We argue that the same effect applies to the majority of pest species due to the correlation of their demography with that of humans. PMID- 11420374 TI - The inference of stepwise changes in substitution rates using serial sequence samples. AB - It is frequently true that molecular sequences do not evolve in a strictly clocklike manner. Instead, substitution rate may vary for a number of reasons, including changes in selection pressure and effective population size, as well as changes in mean generation time. Here we present two new methods for estimating stepwise changes in substitution rates when serially sampled molecular sequences are available. These methods are based on multiple rates with dated tips (MRDT) models and allow different rates to be estimated for different intervals of time. These intervals may correspond to the sampling intervals or to a priori--defined intervals that are not coincident with the times the serial samples are obtained. Two methods for obtaining estimates of multiple rates are described. The first is an extension of the phylogeny-based maximum-likelihood estimation procedure introduced by Rambaut. The second is a new parameterization of the pairwise distance least-squares procedure used by Drummond and Rodrigo. The utility of these methods is demonstrated on a genealogy of HIV sequences obtained at five different sampling times from a single patient over a period of 34 months. PMID- 11420375 TI - Widespread distribution of extensive chromosomal fragmentation in ciliates. AB - Ciliates are a diverse group of eukaryotes characterized by their division of nuclear function into a "germ line" micronucleus and a "somatic" macronucleus. After conjugation, chromosomes in the transcriptionally active macronucleus develop by fragmentation, elimination, and amplification of germ line chromosomes. Extensive chromosomal processing that generates a macronucleus with gene-sized fragments has thus far been well documented in members of only one class of ciliates, the Spirotrichea. Here we establish the broad distribution of extensive fragmentation among members of the class Phyllopharyngea and the genera Metopus (order Armophorida) and Nyctotherus (order Clevelandellida). Moreover, analyses of small-subunit rDNA genealogies indicate that gene-sized chromosomes occur in members of the three separate clades: (1) the class Spirotrichea, (2) the class Phyllopharyngea, and (3) the two orders Clevelandellida and Armophorida. Together, these data indicate that the generation of gene-sized chromosomes is widespread and demonstrate multiple origins of extensive fragmentation within ciliates. PMID- 11420376 TI - Bacterial origin for the isoprenoid biosynthesis enzyme HMG-CoA reductase of the archaeal orders Thermoplasmatales and Archaeoglobales. AB - The enzyme 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase or HMGR) fulfills an essential role in archaea, as it is required for the synthesis of isoprenoid ethers, the main component of archaeal cell membranes. There are two clearly homologous but structurally different classes of the enzyme, one found mainly in eukaryotes and archaea (class 1), and the other found in bacteria (class 2). This feature facilitated the identification of several cases of interdomain lateral gene transfer (LGT), in particular, the bacterial origin for the HMGR gene from the archaeon Archaeoglobus fulgidus. In order to investigate if this LGT event was recent and limited in its scope or had a broad and long term impact on the recipient and its related lineages, the HMGR gene was amplified and sequenced from a variety of archaea. The survey covered close relatives of A. fulgidus, the only archaeon known prior to this study to possess a bacterial-like HMGR; representatives of each main euryarchaeal group were also inspected. All culturable members of the archaeal group Archaeoglobales were found to display an HMGR very similar to the enzyme of the bacterium Pseudomonas mevalonii. Surprisingly, two species of the genus Thermoplasma also harbor an HMGR of bacterial origin highly similar to the enzymes found in the Archaeoglobales. Phylogenetic analyses of the HMGR gene and comparisons to reference phylogenies from other genes confirm a common bacterial origin for the HMGRs of Thermoplasmatales and Archaeoglobales. The most likely explanation of these results includes an initial bacteria-to-archaea transfer, followed by a another event between archaea. Their presence in two divergent archaeal lineages suggests an important adaptive role for these laterally transferred genes. PMID- 11420377 TI - Predicting evolutionary potential. I. Predicting the evolution of a lactose-PTS system in Escherichia coli. AB - Genomes contain not only information for current biological functions, but also information for potential novel functions that may allow the host to adapt to new environments. The field of experimental evolution studies that potential by selecting for novel functions and deducing the means by which the function evolved, but until now it has not attempted to predict the outcomes of such experiments. Here I present a model system that is being developed specifically to examine the issue of what kind of information is most useful in predicting how novel functions will evolve. The system is the evolution of a Lac-PTS transport system and a phospho-beta-galactosidase hydrolase system as a novel pathway for metabolism of lactose in Escherichia coli. Two kinds of information, sequence based phylogenetic inference and biochemical activity, are considered as predictors of which E. coli genes will evolve the required new functions. Both biochemical data and phylogenetic inference predict that the cryptic celABC genes, which currently specify a PTS-beta-glucoside transport system, are most likely to evolve into a PTS-lactose transport system. Phylogenetic inference predicts that the bglA gene, which currently specifies a phospho-beta glucosidase, is most likely to evolve into a phospho-beta-galactosidase. In contrast, biochemical data predict that the cryptic bglB gene, which also currently specifies a phospho-beta-glucosidase, is most likely to evolve into a phospho-beta-galactosidase. PMID- 11420378 TI - Fungal Zuotin proteins evolved from MIDA1-like factors by lineage-specific loss of MYB domains. AB - Proteins are often characterized by the presence of multiple domains, which make specific contributions to their cellular function. While the gain of domains in proteins by duplication and shuffling is well established, domain loss is poorly documented. Here, we provide evidence that domain loss has played an important role in the evolution of protein architecture and function by demonstrating that fungal Zuotin proteins evolved from MIDA1-like proteins, present in animals and plants, by complete loss of the carboxyl-terminal MYB domains. Phylogenetic analyses of the DnaJ motif (the J domain) present in both Zuotin and MIDA1 proteins were complicated by the limited length and profound differences in evolutionary rates exhibited by this domain. To rigorously examine J domain phylogeny, we combined the nonparametric bootstrap with Monte Carlo simulation. This method, which we have designated the resampled parametric bootstrap, allowed us to assess type I and type II error associated with these analyses. These results revealed significant support for domain loss rather than domain gain or gene loss involving paralogs. The absence of sequences related to the MIDA1 MYB domains in Saccharomyces cerevisiae further indicates that the domains have been completely lost, consistent with known functional differences between Zuotin and MIDA1 proteins. These analyses suggest that the description of additional examples of complete domain loss may provide a method to identify orthologous proteins exhibiting functional differences using genomic sequence data. PMID- 11420379 TI - Complete mitochondrial genome sequence of the polychaete annelid Platynereis dumerilii. PMID- 11420380 TI - The putative glutamate receptors from plants are related to two superfamilies of animal neurotransmitter receptors via distinct evolutionary mechanisms. PMID- 11420381 TI - The power to detect recombination using the coalescent. PMID- 11420382 TI - Characterization of the genomic organization, localization and expression of four PRY genes (PRY1, PRY2, PRY3 and PRY4). AB - PRY (PTP-BL related on the Y chromosome) has been proposed as a candidate spermatogenesis gene. We report the characterization of the genomic structure, the number of copies on the Y chromosome and the expression of the gene. By comparison of the cDNA sequence with the genomic sequence, five exons were identified. Analysis of GenBank-derived clones on the Y chromosome revealed the presence of two full-length copies in azoospermia factor region b (AZFb) (PRY1 and PRY2) and two shorter versions of the PRY gene containing exons 3, 4 and 5 in AZFc (PRY3 and PRY4). A clone containing sequences homologous to exons 3, 4 and 5 is located in area 5L (between AZFa and AZFb), a clone containing a sequence homologous to exon 5 is located in area 5M (in AZFb) and a clone containing a fragment homologous to exon 3 is located in 6F. A repeat structure of exons 1 and 2 is present on the short arm of the Y chromosome as well as on the long arm. PRY1 and PRY2, two gene copies that are located in AZFb, a region often deleted in patients with severe male infertility, were shown to be expressed in the testis. PRY may therefore play an important role in spermatogenesis. PMID- 11420383 TI - Expression of the transmembrane carbonic anhydrases, CA IX and CA XII, in the human male excurrent ducts. AB - Testicular fluid is concentrated and acidified during its passage through the excurrent ducts. These processes involve bicarbonate absorption, in which carbonic anhydrases are implicated. In this study, the distribution of two transmembrane carbonic anhydrase isozymes (CA IX and CA XII) in the human excurrent ducts was investigated using isozyme-specific antibodies in conjunction with immunohistochemical and immunoblotting techniques. Specific staining for CA XII was present in the basolateral plasma membrane of the epithelial cells in the efferent ducts, predominantly in the non-ciliated cells. In the epididymal duct, CA XII was detected only in sporadic cells, which also contained CA II, thus suggesting that they are apical mitochondria-rich cells. CA IX was also localized to the basolateral plasma membrane of the epithelium in the efferent ducts, but its staining was weaker and less uniform compared to CA XII. No signal for CA IX was detected in the epididymal duct. Western blot analysis from efferent duct samples revealed specific bands for CA IX and CA XII, confirming that the immunohistochemical stainings represent these isozymes. The expression of CA XII and CA IX in the excurrent duct system and co-expression of CA XII with Aquaporin 1 in the same efferent duct epithelial cells suggest their functional involvement in ion transport and concentration processes of testicular fluid. PMID- 11420384 TI - Changes of the major sperm maturation-associated epididymal protein HE5 (CD52) on human ejaculated spermatozoa during incubation. AB - HE5 (CD52) is a glycoprotein which is secreted by the epididymis and which becomes inserted onto maturing spermatozoa. We have previously shown that, in cynomolgus monkey spermatozoa, changes occur upon maturation rendering cryptic the epitope to the monoclonal antibody CAMPATH-1G; the recognition site is then re-exposed during incubation under capacitation conditions. The present study investigated human ejaculated spermatozoa during incubation under similar conditions, using monoclonal antibodies that recognize different epitopes of the HE5 molecule comprising parts of the N-glycan (2E5) or peptide segments, including (CAMPATH-1G) or excluding (097) the glycosylphosphatidylinositol (GPI) anchor, to reveal modifications of sperm surface HE5. Flow cytometric analysis showed equally high percentages (approximately 90%) of viable spermatozoa cross reacting with the antibodies before and after 6 h incubation. However, during incubation, the staining intensity increased 57% with CAMPATH-1G, 31% with 097, but remained unchanged with 2E5. The lymphocyte CD52 antibody CF1D12 stained only approximately 10% of spermatozoa either before or after incubation. Western blotting of sperm protein extracts using lectins indicated an increase in the exposure of sialic acid residues of HE5 after incubation. These results suggest that during incubation in capacitating conditions, there is an opening up of the HE5 glycoprotein molecule, increasing accessibility of some sialic acid residues and of the core peptide, particularly the GPI anchor. PMID- 11420385 TI - Cloning and sequencing of SOB3, a human gene coding for a sperm protein homologous to an antimicrobial protein and potentially involved in zona pellucida binding. AB - We have previously characterized an 18-19 kDa cationic protein, SOB3, that was detected in the epididymis and localized within the acrosome and on the neck region of human spermatozoa. We suggested that it is involved in secondary sperm binding to the zona pellucida. The present study describes its purification to homogeneity by preparative electrophoresis and non-equilibrium pH gradient electrophoresis. Degenerate primers deduced from microsequencing were used to amplify a specific fragment from human epididymal RNA by reverse transcription polymerase chain reaction (RT-PCR). This 164 bp fragment was extended by 5' and 3'-RACE to obtain the 548 bp full length cDNA. The open reading frame encodes a 170 amino acid protein. SOB3 is a single copy gene. It is 98% identical to prepro FALL39 and 100% identical to CAP18, two human genes which were initially identified by screening a human bone marrow (lambda)gt11 library, and which encode an antimicrobial protein. Northern blots of human tissues revealed a 1 kb transcript in corpus and cauda epididymis only, while RT-PCR showed presence of the mRNA in the three epididymal regions and also in round spermatids. The above results suggest that SOB3 has two roles in sperm protection and fertilization, depending on its dual origin and final sperm localization. PMID- 11420386 TI - Anti-SLIP1-reactive proteins exist on human spermatozoa and are involved in zona pellucida binding. AB - Sulpholipid immobilizing protein 1 (SLIP1) is an evolutionarily conserved 68 kDa plasma membrane protein, present selectively in germ cells. We have previously shown that mouse sperm SLIP1 is involved in sperm-zona pellucida (ZP) binding. In this report, we extended our study to the human system. Immunoblotting demonstrated that anti-SLIP1-reactive proteins (mol. wt 68 and 48 kDa) could be extracted from human spermatozoa by an ATP-containing solution, a result that is consistent with observations in other species. Direct immunofluorescence, using Cy3-conjugated anti-SLIP1 IgG, revealed SLIP1 staining over the acrosomal region, with higher intensity at the posterior area. Using the human sperm-ZP binding assay, we demonstrated that pretreatment of human spermatozoa from three donors with anti-SLIP1 IgG revealed lower numbers of zona-bound spermatozoa, as compared to the corresponding control spermatozoa treated with normal rabbit serum IgG. This decrease in zona pellucida binding was not from an antibody-induced decline in sperm motility or an increase in the premature acrosome reaction. The results strongly suggest that anti-SLIP-reactive proteins on human spermatozoa play an important role in ZP binding. PMID- 11420387 TI - Matrix metalloproteinases and tissue inhibitors of metalloproteinases in human fetal testis and ovary. AB - Matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) are major regulators of tissue remodelling of the extracellular matrix (ECM) and may also be involved in the control of growth factor availability. We have investigated their production and localization in the developing human gonad during mid-gestation using zymographic techniques and immunohistochemistry. The secretion of MMP-2, MMP-9 and all four TIMP was demonstrated from both testis and ovary, with the predominant gelatinase produced by both being MMP-2. In the testis, MMP-1, MMP-2, MMP-9 and all TIMP family members were localized to the interstitium and to varying degrees within the tubules. MMP-9 and TIMP-4 were abundant in both Sertoli cells and gonocytes and MMP-1 and TIMP-1 were localized in particular to Sertoli cells. In the ovary, all TIMP and MMP-1, MMP-2 and MMP-9 were localized to the oogonium/oocyte cytoplasm with varying intensities and MMP 1, TIMP-2 and TIMP-3 were also detected in the ovarian stroma. This study demonstrates that MMP-1, MMP-2, MMP-9 and all TIMP family members are secreted by the developing ovary and testis and are localized to specific cell and tissue sites. MMP and TIMP are likely to play a role in ECM remodelling during gonadal development and also in the cell and matrix interactions that control a range of cellular functions. PMID- 11420388 TI - Evidence for the expression of interleukin (IL)-18, IL-18 receptor and IL-18 binding protein in the human endometrium. AB - To see whether the interleukin (IL)-18 system is operative in the endometrium, we examined the expression of IL-18, IL-18 receptor (IL-18R) and IL-18 binding protein (IL-18BP), the substance known to neutralize IL-18 activity, in this tissue. Reverse transcription-polymerase chain reaction analyses showed that IL 18, IL-18R and IL-18BP mRNA were constitutively expressed without significant fluctuation throughout the menstrual cycle. When epithelial cells and stromal cells were cultured separately, the expression levels of IL-18 mRNA in epithelial cells were about 18-fold higher compared to those in stromal cells. Furthermore, the IL-18 precursor protein was detected by Western blot analysis in cultured epithelial cells but not in stromal cells. Recombinant human IL-18 stimulated the secretion of interferon (IFN)-gamma by resident bone marrow-derived cells in the endometrium. On the other hand, IFN-gamma up-regulated the IL-18BP expression both in cultured epithelial cells and stromal cells. Thus, we have presented evidence for the presence of the IL-18 system in the human endometrium. In light of its immunomodulatory roles in a variety of tissues, this system may afford protection against pathogenic micro-organisms and provide a regulatory mechanism for controlled trophoblast invasion by modulating a local cytokine network. PMID- 11420389 TI - Differential and cell-specific expression of calcitonin receptor-like receptor and receptor activity modifying proteins in the human uterus. AB - The calcitonin receptor-like receptor (CRLR) can function as a receptor for either calcitonin gene-related peptide (CGRP) or adrenomedullin (AM), depending upon co-expression with members of a novel family of receptor activity-modifying proteins (RAMP). RAMP1 presents the CRLR at the cell surface as a CGRP/AM receptor. RAMP2- and RAMP3-transported CRLR receptors act as AM-specific receptors. However, it is still unknown if this signalling system operates in vivo. Of particular interest is the uterus, where both peptides and their binding sites are known to be present and where both mitogenic and vasodilatory responses to AM and CGRP have been demonstrated. In this study, we examined whether CRLR and RAMP are co-expressed in the same populations of cells in human uterine tissue. Analysis by in-situ hybridization and immunocytochemistry revealed a heterogeneous and cell type-specific distribution of components of this AM/CGRP signalling system. Adrenomedullin mRNA was expressed and evenly distributed across all cell types. CRLR mRNA was predominantly found in blood vessels. RAMP1 expression was specific to myometrial myocytes and vascular smooth muscle cells in uterine arteries. RAMP2 and RAMP3 mRNA were not detectable by in-situ hybridization. The pattern of differential and cell-specific expression of CRLR and RAMP suggests the involvement of CRLR/RAMP1 in the processes of vasodilation, smooth muscle relaxation and angiogenesis in response to AM and CGRP in the human uterus, but also indicates that other receptors may be implicated. PMID- 11420390 TI - Menstrual cycle-specific inhibition of endometrial stromal cell proliferation by oncostatin M. AB - We have investigated the possible roles of oncostatin M (OSM), which is a member of the interleukin-6 family of cytokines, in endometrial and endometriotic stromal cell growth. Endometrial and endometriotic stromal cells were collected from the uterus or ovarian chocolate cysts. We observed the expression of mRNA transcripts for OSM, OSM receptor subunit beta, leukaemia inhibitory factor receptor subunit (LIFR), and glycoprotein 130 in endometrial and endometriotic stromal cells. We also examined the effects of OSM (0-50 ng/ml) and LIF (0-10 ng/ml) on endometrial and endometriotic stromal cell proliferation and evaluated the effects of OSM on endometrial stromal cell differentiation. The presence of 10-50 ng/ml OSM significantly suppressed endometrial stromal cell growth in secretory phase tissue but not in proliferative phase tissue. In contrast, stromal cells in endometriotic tissues were resistant to the inhibitory effects of OSM. Addition of LIF did not influence the growth of endometrial stromal cells. We also showed that 10 ng/ml OSM stimulated markers of differentiation causing increased prolactin secretion and cyclooxygenase-2 gene expression in endometrial stromal cells from the secretory phase. These results suggest that OSM may play a pivotal role in regulating the growth and differentiation of endometrial cells. Endometriotic cells may behave differently from normal endometrial cells in terms of the inhibitory response to OSM. PMID- 11420391 TI - Oestriol and oestradiol increase cell to cell communication and connexin43 protein expression in human myometrium. AB - Oestradiol increases the protein expression of connexin43 (Cx43) gap junctions in myometrium but the effect of oestriol on gap junction expression has not been described previously. Oestriol is the most abundant free oestrogen in pregnant women and there is a marked surge in oestriol concentrations before term and idiopathic preterm labour. In order to determine whether oestriol may have a physiological action on the myometrium, cultured human myometrial cells obtained from non-pregnant hysterectomy specimens were exposed to 10 nmol/l oestradiol or oestriol. Intercellular communication between myometrial cells was investigated by microinjection of confluent cultured cells with the gap junction-permeant tracer Cascade Blue. There was a progressive increase in coupling after exposure to oestradiol or oestriol (P < 0.0005). An increase in Cx43 protein expression was demonstrated by immunocytochemistry after 1 h (P < 0.01) and 3 days (P < 0.01) exposure, and by Western blotting after 1 h (P < 0.01) and 3 days (P < 0.05) exposure, to both oestradiol and to oestriol. We conclude that oestriol increases gap junction communication in human myometrium by increasing gap junction expression. Elevated oestriol concentrations may thus play a role in the initiation of labour in women, by increasing cell-cell communication in the myometrium. PMID- 11420392 TI - Apoptosis in the normal human amnion at term, independent of Bcl-2 regulation and onset of labour. AB - This study was designed to detect apoptosis in the human amnion and to elucidate the signalling pathway involved in its regulation. Samples of human amnion were obtained from 34 women (weeks 11-42 of gestation) and studied using the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL) method with light microscopy (LM) and transmission electron microscopy (TEM). Apoptotic regulators in the samples were studied by immunohistochemistry and caspase activity assay. The TUNEL method with LM demonstrated that the percentage of TUNEL-positive cells in the amniotic epithelium was the highest in weeks 40-41 of gestation (P < 0.05) independent of the onset of labour, and the cells were often detached from the epithelium into the amniotic cavity at term. The TUNEL method with TEM clearly showed the characteristic features of apoptosis such as the nuclear condensed chromatin with abundant free 3'-OH DNA ends, cell shrinkage and a decrease in the number of desmosomes, except for the presence of apoptotic bodies. Fas and Fas ligand (FasL) were constantly expressed on apical membranes of amniotic epithelial cells from weeks 16-27 through to 40-41 of gestation, while no Bcl-2 expression was observed throughout the gestational periods. Activities of caspase-3 and caspase-8, but not of caspase-9, were higher in weeks 40-41 than those from weeks 16-27 of gestation (P < 0.01). We conclude that apoptosis in term amniotic epithelium is independent of Bcl-2 regulation and onset of labour, and may play an important role in the fragility and rupture of human fetal membranes at term. PMID- 11420393 TI - The role of USP9Y and DBY in infertile patients with severely impaired spermatogenesis. PMID- 11420395 TI - Radiation-induced chromosome aberrations in human euchromatic (17cen-p53) and heterochromatic (1cen-1q12) regions. AB - The constitutively heterochromatic 1q12 band and the primarily euchromatic 17cen p53 region comprise a similar size in terms of percentage of the total human genome but have a completely distinguishable chromatin structure. The aim of this study is to unravel whether this structural difference has an impact on the formation and processing of radiation-induced chromosome aberrations. To do so, we have analysed the initial induction and the long-term persistence of radiation induced (3 Gy gamma-rays) chromosomal aberrations with breakpoints in either the 1q12 band or the 17cen-p53 region in comparison with the behaviour of the overall genome. The fusigenic potential of euchromatic and heterochromatic ends was also compared. This time course experiment was performed in a human lymphoblastoid cell line with sampling times at 1, 3, 7, 14 and 56 days after irradiation. The outcome of this study, with 68 000 metaphases studied by multicolour FISH, with centromeric (1cen and 17cen), paracentric (1q12) and locus specific (p53 gene) probes, revealed: (i) a similar radiosensitivity of all regions analysed irrespective of their chromatin configuration; (ii) a possible enhanced fusigenic potential of heterochromatic chromosome ends; (iii) a rapid decline of 1q12 translocations; and (iv) a similar long-term behaviour of translocations involving 1q12 and 17cen-p53. The implications of these findings in biomonitoring studies are discussed. PMID- 11420396 TI - Inter-individual differences in repair of DNA base oxidation, measured in vitro with the comet assay. AB - There is a need for a reliable, robust and sensitive assay for DNA repair, suitable for use with human lymphocyte samples in molecular epidemiological investigations. The comet assay (single cell alkaline gel electrophoresis) has been modified to measure the ability of a simple subcellular extract of lymphocytes to carry out the initial step of repair, i.e. incision, on a DNA substrate carrying specific lesions--namely, oxidized bases introduced by visible light in the presence of photosensitizer. The cell extract is free of non specific nuclease activity, incising DNA only if the DNA has been treated with photosensitizer and light. The activity varies between individuals, but consistency is seen between samples from each individual taken on occasions several months apart. The lack of activity of extract from Ogg1(-) mouse cells (deficient in the glycosylase that excises 8-oxoguanine) in this assay confirms that the activity measured is predominantly excision repair of oxidized bases. This new DNA repair assay is simple, rapid and requires only small quantities of lymphocyte extract (obtainable from 10 ml blood). PMID- 11420397 TI - Mutagenic potential of bisphenol A diglycidyl ether (BADGE) and its hydrolysis derived products in the Ames Salmonella assay. AB - The mutagenicity of bisphenol A diglycidyl ether (BADGE), its first and second hydrolysis products (the diol epoxide and bis-diol of BADGE, respectively) and the bis-chlorohydrin of BADGE were investigated using the Ames Salmonella assay with strains TA98, TA100, TA1535 and TA1537. The assays were performed in the absence and presence of various concentrations of rat liver S9 fraction. The results obtained confirm the mutagenic power of BADGE in strains TA100 and TA1535 and show a positive response to the diol epoxide of BADGE in these strains, although the latter compound was approximately 10 times less potent than the former. A lack of mutagenic activity of the bis-diol of BADGE and the chlorohydrin under study is also shown. These findings suggest that BADGE and, to a much lesser extent, the diol epoxide of BADGE may constitute a genotoxic hazard, but not the bis-diol or bis-chlorohydrin of BADGE. PMID- 11420398 TI - Intestinal tumours induced by the food carcinogen 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine in multiple intestinal neoplasia mice have truncation mutations as well as loss of the wild-type Apc(+) allele. AB - C57BL/6J-Min/+ (multiple intestinal neoplasia) is a murine model for familial adenomatous polyposis (FAP), where the mice are heterozygous for a nonsense Apc(Min) (adenomatous polyposis coli) mutation, and therefore develop numerous spontaneous adenomas in the small intestine and colon. Neonatal exposure of Min/+ mice to the food carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) (eight subcutaneous injections of 25 or 50 mg/kg PhIP to pups or 50 mg/kg PhIP to lactating dams) markedly increased (2--9-fold) the number of intestinal tumours, especially in the small intestine. We examined whether the Apc gene was affected in small intestinal and colonic tumours induced by PhIP. In spontaneous tumours formed in these mice, the main mechanism for tumour induction is loss of the wild-type Apc(+) allele, i.e. loss of heterozygosity (LOH). Also in the PhIP induced tumours, this is a major mechanism, since large fractions of PhIP-induced tumours had LOH in APC: However, mechanisms other than LOH must also prevail, since a lower frequency of LOH was found in the small intestinal tumours from male mice exposed to PhIP either via breast milk (65%) or by direct injection (68%), compared with the untreated controls (92%). Tumours that had retained the wild-type Apc(+) allele were further analysed for presence of truncated Apc proteins with in vitro synthesized protein (IVSP) assay. Truncated Apc proteins, indicating truncation mutations in exon 15 of the Apc gene, were detected in 20% (8 of 40) of the tumours not showing LOH from the small intestine after PhIP exposure, all in segment 2 (codons 686--1217). Seventeen percent (2 of 12) of the colonic tumours had a truncated Apc protein in segment 3 (codons 1099--1693). Importantly, no truncated proteins were detected in tumours from unexposed mice with apparently retained wild-type Apc(+) allele. These results show that PhIP induces intestinal tumours in the Min/+ mice both by causing LOH and truncation mutations in the wild-type Apc(+) allele. PMID- 11420399 TI - Normal human lymphocytes exhibit a wide range of methionine-dependency which is related to altered cell division but not micronucleus frequency. AB - The underlying cause(s) of methionine-dependency and its relevance to cancer remains unclear. We aimed to determine whether (i) normal human lymphocytes exhibit methionine-dependency, (ii) baseline levels of genetic damage are related to methionine-dependency and (iii) methionine-dependence can be explained, in part, by common polymorphisms in methionine synthase and methylenetetrahydrofolate reductase (MTHFR). Genetic damage was measured in lymphocytes of 52 volunteers (29--65 years) using the cytokinesis-block micronucleus assay. Methionine-dependency was assessed by culturing cells in serum-free media containing 0.1 mM L-methionine and 0 mM D,L-homocysteine (met(+)hcy(-)) or 0 mM L-methionine and 0.2 or 0.4 mM D,L-homocysteine (0.2/0.4 hcy(+))(met(-)hcy(+)). Mitogenesis was stimulated with phytohaemagglutinin. Cytokinesis was inhibited by adding cytochalasin B at 44 h. Ninety-six hours after PHA, cells were transferred to microscope slides. Cell proliferation was measured by counting binucleated cell frequency and calculating nuclear division index. Volunteers were classified into tertiles of methionine-dependence according to the growth of their cells in met(-)hcy(+) media (relative to growth in met(+)hcy(-) media). Average cell division, as a percentage of division in met(+)hcy(-) media, was approximately 5, 26 and 70% in 0.2-hcy(+) media and 29, 70 and 142% in 0.4-hcy(+) media for the high, mid and low tertiles of methionine dependence, respectively. Micronucleus frequency did not vary between these tertiles (P > 0.6). In both met(+)hcy(-) and met(-)hcy(+) media, cell division was not affected by polymorphisms in MTHFR (C677T, A1298C) or methionine synthase (A2756G). Cell division in met(-)hcy(+) media was negatively correlated with division in met(+)hcy(-) media (P = 0.05 and 0.007 for 0.2 and 0.4-hcy(+), respectively). Methionine-dependent lymphocytes had higher levels of cell proliferation in met(+)hcy(-) media than methionine-independent lymphocytes (P = 0.089 and 0.01 for 0.2 and 0.4-hcy(+), respectively). However, this difference was not apparent in previous experiments when cells were grown in media containing 10% fetal calf serum. These findings show that there is a wide inter individual variation in the degree of methionine-dependency of normal human lymphocytes in vitro. Methionine-dependency does not appear to alter the risk for chromosomal mutation as measured by the micronucleus assay. We discuss the possible relevance to cancer of increased cell division in methionine-dependent cells under methionine-replete and serum-free media conditions. PMID- 11420400 TI - Spontaneous and induced chromosome damage in somatic cells of sporadic and familial Alzheimer's disease patients. AB - Alzheimer's disease (AD) is a neurodegenerative disorder of the elderly with a complex etiology due to the interaction between genetic and environmental factors. At least 15% of cases are inherited as an autosomal dominant mutation, but the majority are sporadic. We evaluated cytogenetic alterations, both spontaneous and chemical-induced [aluminium (Al) and griseofulvin (GF)], by means of the micronucleus (MN) test in lymphocytes or skin fibroblasts of 14 patients with sporadic and eight with familial Alzheimer's disease (FAD), respectively. The spontaneous MN frequencies of sporadic (20.8 +/- 9.2) and familial (20.7 +/- 4.6) AD patients are significantly higher than those of the respective control groups (9.0 +/- 6.8 and 6.7 +/- 3.4). In all AD patients, GF significantly increased the spontaneous MN frequency of somatic cells to a lesser extent (P < 0.05) as compared with the control group. Al treatment did not induce MN in AD patients. The results of the present study indicate that different types of somatic cells from sporadic and familial AD patients show comparable levels of spontaneous cytogenetic anomalies, and MN induction is partially reduced or lacking according to the type of chemical treatments. PMID- 11420401 TI - The male rat carcinogens limonene and sodium saccharin are not mutagenic to male Big Blue rats. AB - Limonene and sodium saccharin are male rat specific carcinogens giving rise to renal and bladder tumours, respectively. Both compounds give negative results in genetic toxicity assays suggesting a non-genotoxic mode of action for their carcinogenicity. The alpha 2U-globulin accumulation theory has been invoked to explain the renal carcinogenicity of limonene: the accumulation of micro masses of calcium phosphate in the bladder, coupled with a high pH environment in the male rat bladder, has been suggested to be responsible for the bladder carcinogenicity of sodium saccharin. The implication of these proposed mechanisms is that limonene and sodium saccharin will not be mutagenic to the rat kidney and bladder, respectively. This proposal has been evaluated by assessing the mutagenic potential of the two chemicals to male lacI transgenic (Big Blue) rats. Male Big Blue rats were exposed for 10 consecutive days to either limonene in diet, at a dose level in excess of that used in the original National Toxicology Program gavage carcinogenicity bioassay, or to sodium saccharin in diet at the dose known to induce bladder tumours. The multi-site rat carcinogen 4 aminobiphenyl was used as a positive control for the experiment. Limonene failed to increase the mutant frequency in the liver or kidney of the rats, and sodium saccharin failed to increase the mutant frequency in the liver or bladder of the rats. 4-Aminobiphenyl was mutagenic to all three of these tissues. These results add further support to a non-genotoxic mechanism of carcinogenic action for both limonene and sodium saccharin. PMID- 11420402 TI - Studies of dose distribution, premutagenic events and mutation frequencies for benzo[a]pyrene aiming at low dose cancer risk estimation. AB - Cancer risk assessment of polycyclic aromatic hydrocarbons (PAH) is complicated by several of these compounds exerting a promoter action leading to high tumour incidences at high doses. Cancer risks at low doses corresponding to the uptake from air and food in the general environment would best be estimated on the basis of measurement of in vivo target doses of genotoxic (mutagenic) intermediates and a determination of mutation frequency per unit of dose. In experiments ultimately aiming at a risk assessment of environmental PAH from in vivo doses benzo[a]pyrene (BaP) was chosen as a model. gamma-Radiation has earlier been used as a reference standard in cancer risk estimation of genotoxic chemicals where dose equivalents (rad-equivalents) have been shown to give reliable risk estimates for several alkylating agents. Variation in dose of BaP diolepoxide between organs was studied by measurement of deoxyguanosine-N(2) adducts in DNA after administration of BaP by gavage to mice of a strain with reduced DNA repair (Xpa(-/-)). The adduct levels in spleen, forestomach, stomach and small intestine were approximately the same; with the adduct level in spleen as reference it was twice as high in liver and lung and about half as high in colon tissue. A chemical or radiation dose is proportional to the cumulative frequency of putatively premutagenic changes (premutagenic hits) in DNA. The mutation frequency per premutagenic hit (genotoxic chemicals) and per unit of dose (gamma radiation) were calculated from acutely exposed V79 cells in order to determine the mutagenic effectiveness of each agent. Based on the mutagenic effectiveness determined in this study 10(-4) Gy can be regarded equally effective in causing phenotypically expressed HPRT mutations as the dose of BaP which causes the formation of one deoxyguanosine-N(2) adduct per cell. PMID- 11420403 TI - Effect of chemicals on the duration of male meiosis in mice detected with laser scanning cytometry. AB - Aneuploidy studies in sperm such as the sperm-FISH assay require a precise knowledge of the duration of spermatogenesis, especially of the meiotic stages. This is important in order to sample sperm from the epididymis at appropriate intervals after animal treatment. However, aneugens may delay the cell cycle. The progression from meiotic divisions to epididymal sperm was determined by labelling the last S-phase before meiosis with the thymidine analogue 5-bromo-2' deoxyuridine (BrdU) and treating the animals 13 days later with the test chemicals. In a time frame of 20--24 days after treatment, BrdU-containing sperm were identified with a FITC-labelled anti-BrdU antibody and green fluorescent sperm were scored with a laser scanning cytometer (LSC). We studied the effects of the chemicals acrylamide, colchicine, diazepam, griseofulvin, taxol, thiobendazole, trichlorfon and vinblastine on the duration of meiotic divisions in male mice. Colchicine treatment prolonged the duration of meiotic divisions by about 48 h. On days 21 and 22, the frequencies of BrdU-labelled sperm in the colchicine group were 11.7 and 9.4%, respectively, while they were 28.4 and 30.6%, respectively, in the concurrent controls (P > 0.01). On day 24 after treatment, the frequency of labelled sperm in the colchicine group reached the control level. Etoposide treatment resulted in an elevation of BrdU-labelled sperm at 23 rather than 22 days. The other chemicals showed no significant effect of prolonging meiotic cell cycle progression. On the basis of the colchicine and etoposide data, it is suggested that the effect of a chemical on the meiotic cell cycle progression is determined first in order to chose the appropriate sperm sampling time to detect aneuploidy induction. PMID- 11420404 TI - Evaluation of the cytogenetic damage induced by the antihypertensive drug nimodipine in human lymphocytes. AB - The aim of this work was a study of the genotoxic potential of chronic long-term therapy with the antihypertensive drug nimodipine by measures of sister chromatid exchanges (SCE) and micronuclei (MN) in peripheral human lymphocytes of patients with long-term exposure to this drug. Peripheral human lymphocytes of control individuals exposed in vitro to nimodipine were also studied to assess the effect of the drug itself. Fluorescence in situ hybridization (FISH) with a centromeric probe was performed to determine the origin of the induced MN. The in vivo study was carried out on five patients under antihypertensive treatment with nimodipine. The in vitro study was performed on five control individuals by adding the drug to the culture medium at a final concentration similar to the levels found in plasma (controls/medium). The in vivo study showed no genotoxic effects of long-term therapy with nimodipine because the frequencies of SCE and MN in exposed patients did not show significant differences as compared with control individuals. A statistically significant increase in the frequency of MN was detected in controls/medium as compared with control individuals without the drug. FISH analysis revealed statistically significant differences with respect to the frequency of centromeric signals in nimodipine-induced MN in vitro. With regard to the in vivo results, chronic long-term therapy with nimodipine is not associated with increased genotoxicity. The differing results in vivo and in vitro could be due to extensive metabolism of nimodipine, indicating that the cytogenetic effect observed was due to the drug itself rather than its metabolites or to an adaptive response to nimodipine in vivo. PMID- 11420405 TI - Time- and dose-dependent DNA binding of PAHs derived from diesel particle extracts, benzo[a]pyrene and 5-methychrysene in a human mammary carcinoma cell line (MCF-7). AB - Cultures of a human mammary carcinoma cell line (MCF-7) were exposed to the soluble organic fraction of diesel particle emissions, benzo[a]pyrene (B[a]P) and 5-methylchrysene (5-MeCHR) to study time- and dose-related PAH-DNA binding. The concentrations of 14 PAHs in three extracts were analyzed by HPLC and PAH-DNA adducts were measured by (32)P post-labeling assay. Time-dependent DNA adducts formation of 2.5 microM B[a]P was lower than that of 2.5 microM 5-MeCHR. In comparison with B[a]P, 2-fold higher adduct formation by 5-MeCHR was observed at 12 h exposure, after which BPDE adducts decreased and 5-MeCHR continued to form adducts linearly during 48 h exposure. The data for these two PAH compounds demonstrate a large variation in adduct-forming potency, which should be taken into account when estimating DNA adducts formed by mixtures of unknown PAHs. A clear dose-response effect on formation of DNA adducts was obtained for B[a]P and a Standard Reference Material (SRM) of diesel particulate matter. The amount of B[a]P contributed more to total DNA adduct formation by SRM than by three diesel extracts. Thus, no conclusions can be drawn from diesel particle-derived B[a]P as to the adduct-forming potency of other carcinogenic PAHs. There was little change in adduct levels formed by three diesel extracts from 0 to 12 h exposure. Thereafter, the number of adducts formed by RD2 increased more rapidly than those formed by RD1 and EN97. The concentrations of 14 PAHs and adduct levels analyzed at 24 and 48 h did not change in the same proportion between the extracts. Neither could PAH-DNA adduct levels be explained by the sum of strong and weak adduct-forming PAHs analyzed in the extracts. This indicates that other PAHs in the extracts RD1, RD2 and EN97 contributed to adduct formation more than the carcinogenic adduct-forming PAHs analyzed in this study. PMID- 11420406 TI - Chromosomal aberration and single cell gel electrophoresis (Comet) assay in the longitudinal risk assessment of occupational exposure to pesticides. AB - In recent years the use of pesticides in agriculture has been increasing steadily. At present there are more than 1000 chemicals classified as pesticides. Therefore, the widespread use of pesticides and their potential genetic hazard suggests that evaluation of their genotoxicity should be extended using the newer assays now available. In the present study chromosomal aberration analysis and the alkaline single cell gel electrophoresis (Comet) assay were used to evaluate the extent of DNA damage and DNA repair in peripheral blood lymphocytes of subjects employed in pesticide production. In order to determine possible primary genotoxic effects in workers blood samples were taken after an 8 month long period of exposure to a complex mixture of pesticides. To detect the possible occurrence of DNA repair in lymphocytes of the same subjects the second blood sample was taken after an 8 month long period of absence from the pesticide exposure zone. Regardless of the period of sampling, in the exposed group statistically significantly increased numbers of aberrant cells, chromatid and chromosome breaks, acentric fragments and dicentric chromosomes compared with the controls were found. After the workers had spent 8 months out of the pesticide exposure zone the number of aberrant cells and all types of chromatid and chromosome aberrations decreased significantly compared with sampling after the high exposure period, but it still remained significantly higher in comparison with the control group. After the period of high exposure to a mixture of pesticides statistically significantly increased levels of DNA damage in the Comet assay in terms of tail length and tail moment were found. After the workers were removed from production for 8 months both Comet assay end-points decreased significantly compared with the first sampling point, but they remained increased compared with the control. PMID- 11420407 TI - Ionizing radiation damage repair: a role for topoisomerases? AB - In parallel with the developing field of DNA topoisomerase poisons in tumor chemotherapy, the basic features of these nuclear enzymes have been unfolded. The role of topoisomerases in fundamental processes involving DNA metabolism has been shown to outpace by far the initial expectations. While DNA topoisomerases are involved in relaxation of chromatin to relieve tension during DNA replication and transcription, as well as for recombinational processes and chromosome segregation and condensation, the possible role, either direct or indirect, of these enzymes in DNA repair is still a matter of discussion. In this survey the possible relationship of topoisomerases with the repair of ionizing radiation damage in mammalian cells is considered, on the basis of attractive 'clues' and in the light of a number of observations. PMID- 11420410 TI - Familial small upper airways and sleep-disordered breathing: relationship to idiopathic apparent-life-threatening events. PMID- 11420411 TI - Should we do lung recruitment maneuvers when giving surfactant? PMID- 11420412 TI - Impaired interferon gamma-mediated immunity and susceptibility to mycobacterial infection in childhood. AB - Mendelian susceptibility to poorly virulent mycobacteria such as bacillus Calmette-Guerin (BCG) and environmental nontuberculous mycobacteria is a clinically heterogeneous syndrome. The clinical features of affected children cover a continuous spectrum from disseminated lethal bacillus Calmette-Guerin infection to local recurrent nontuberculous mycobacterial infection. Different types of mutations in four genes (IFNGR1, IFNGR2, IL12B, IL12RB1) have revealed both allelic and nonallelic heterogeneity and result in eight different disorders whose common pathogenic pathway is impaired interferon gamma (IFNgamma) mediated immunity. The severity of the clinical phenotype depends on the genotype. Complete IL-12 p40 and IL-12 receptor beta1 deficiencies and partial IFNgamma receptor 1 (IFNgammaR1) and IFNgammaR2 deficiencies generally lead to curable infections at various ages, and antibiotics supplemented with IFNgamma if required are likely to be effective. Complete IFNgammaR1 and IFNgammaR2 deficiencies predispose to overwhelming infection in early childhood, which may respond to antibiotics but relapse when antibiotics are discontinued. Rapid discrimination between complete IFNgammaR1 and IFNgammaR2 deficiency and other defects, therefore, is an important diagnostic step for planning clinical management. PMID- 11420413 TI - Sleep-disordered breathing and upper-airway anomalies in first-degree relatives of ALTE children. AB - From 1985 through 1995, 348 infants aged 3 wk-3 mo were referred to the Stanford Sleep Clinic for "apparent life-threatening events" (ALTE). A small group of 48 infants with no history of sleep-disordered breathing (SDB) was also recruited and used as controls (they comprised group C). We conducted a systematic investigation of relatives (parents, siblings, and grandparents) of the infants, including a clinical evaluation, craniofacial investigation, and the completion of an extensive (189-question) validated sleep/wake questionnaire. All data were calculated before the subdivision of ALTE infants into two groups. The subdivision was based on a blind scoring of the infants' polygraphic recordings; 42.5% of the infants were negative for SDB (Group A), whereas 57.5% of the infants were positive for SDB (Group B). Groups A and C were not significantly different from each other. Forty-three percent of the relatives of Group B infants had been treated for SDB (with nasal CPAP, surgical or dental appliance treatments) compared with 7.1% of Group A relatives. Clinical investigation indicated a significantly higher presence of small upper airways in the families of infants with SDB. About twice as many relatives reported the presence of asthma in Group B compared with Group A. Naso-oro-maxillomandibular anatomic traits that may lead to small upper airways in parents may be risk factors for abnormal breathing during sleep in their infants. PMID- 11420414 TI - Inhibition of K(+) channel activity in human pulmonary artery smooth muscle cells by serum from patients with pulmonary hypertension secondary to congenital heart disease. AB - Activity of K(+) channels regulates cytosolic free Ca(2+) concentration by controlling membrane potential. A rise in cytosolic free Ca(2+) concentration in pulmonary artery smooth muscle cells (PASMC) triggers pulmonary vasoconstriction and stimulates PASMC proliferation. Whether serum from children with pulmonary hypertension (PH) secondary to congenital cardiopulmonary diseases contains a factor(s) that inhibits K(+) channel function in PASMC was investigated using patch clamp techniques. PASMC isolated from normal subjects were cultured in media containing 5% serum from normotensive (NPH) or PH patients. Cell growth rate and the currents through voltage-gated K(+) channels were determined and compared between the cells treated with serum from NPH and PH patients. In the absence of growth factors, incubation of PASMC in media containing NPH serum for 48 h increased cell numbers by 2.5-fold, whereas incubation of the cells in media containing PH serum increased cell numbers by 4.5-fold (p < 0.001). Amplitude of whole-cell voltage-gated K(+) currents in NPH serum-treated cells (1119 +/- 222 pA at +80 mV, n = 43) was 3.5-fold greater than in PH serum-treated cells (323 +/ 34 pA, n = 43, p < 0.001). Consistently, membrane potential was much more depolarized in PASMC treated with PH serum (-28 +/- 2 mV, n = 29) than cells treated with NPH-serum (-47 +/- 2 mV, n = 28; p < 0.001). These results suggest that a circulating mitogenic agonist, which induces membrane depolarization by inhibiting voltage-gated K(+) channel activity in PASMC, may be produced or up regulated in pediatric patients with secondary PH. PMID- 11420415 TI - Oxidative stress and increased type-IV collagenase levels in bronchoalveolar lavage fluid from newborn babies. AB - Oxidative stress may increase lung permeability by up-regulation of matrix metalloproteinase-9 (MMP-9), a type-IV collagenase that can disrupt alveolar basement membranes. We have compared a marker of oxidative stress (protein carbonyl residues) with levels of MMP-9 and its inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1), in bronchoalveolar lavage samples from newborn babies. Bronchoalveolar lavage samples (n = 87, two from each time point) were taken in the first 6 postnatal days from 41 ventilated babies: 18 of <29 wk gestation, 10 of 29-36 wk, 9 term with persistent fetal circulation, and 4 term without lung disease. Respiratory disease severity at the time of bronchoalveolar lavage was assessed using the arterial-alveolar oxygen tension ratio. One sample from each time point was used for the measurement of MMP-9 by zymography and TIMP 1 by ELISA. The second sample was used to measure carbonyl group concentrations, also using an ELISA. Correlations were calculated between protein carbonyls, arterial-alveolar oxygen tension ratio, and MMP-9 and TIMP-1 concentrations. Significant correlations were found between carbonyl concentrations and arterial alveolar oxygen tension ratio (r = -0.325, p = 0.0031, n = 81), MMP-9 (r = 0.331, p < 0.0029, n = 79), and TIMP-1 (r = 0.436, p < 0.0001, n = 87). Worsening respiratory disease in newborn babies is associated with increased carbonyl concentrations in neonatal bronchoalveolar lavage fluid, and these correlated with MMP-9 and TIMP-1 levels. Increased oxidative stress may damage the lung by increasing type-IV collagenase activity, causing disruption of the extracellular matrix. PMID- 11420416 TI - Alveolar recruitment promotes homogeneous surfactant distribution in a piglet model of lung injury. AB - Uneven distribution of exogenous surfactant contributes to a poor clinical response in animal models of respiratory distress syndrome. Alveolar recruitment at the time of surfactant administration may lead to more homogeneous distribution within the lungs and result in a superior clinical response. To investigate the effects of three different volume recruitment maneuvers on gas exchange, lung function, and homogeneity of surfactant distribution, we studied 35 newborn piglets made surfactant deficient by repeated airway lavage with warm saline. Volume recruitment was achieved by either a temporal increase in tidal volume or an increase in end-expiratory pressure during surfactant administration, yielding an increase in dynamic compliance of the respiratory system of 77% in the first group and an increase in functional residual capacity of 108% in the second group. A third group of piglets (all n = 7) received a combination of both volume recruitment maneuvers, with increases in dynamic compliance of the respiratory system of 100% and in functional residual capacity of 192%. Those animals subjected to increased tidal volume showed an improved surfactant response in terms of oxygenation, ventilation, lung volumes, lung mechanics, and homogeneity of surfactant distribution. Increased end-expiratory volume augmented the surfactant effect only to some extent. The combination of both volume recruitment maneuvers, however, needed lung volumes beyond total lung capacity (approximately 56 mL/kg), thus probably inducing early sequelae of ventilator-induced lung injury. We conclude that volume recruitment by means of increased tidal volumes at the time of surfactant administration leads to a superior surfactant effect owing to more homogeneous surfactant distribution within a collapsed lung. PMID- 11420417 TI - Resistance of different surfactant preparations to inactivation by meconium. AB - A disease similar to acute respiratory distress syndrome may occur in neonates after aspiration of meconium. The aim of the study was to compare the inhibitory effects of human meconium on the following surfactant preparations suspended at a concentration of 2.5 mg/mL: Curosurf, Alveofact, Survanta, Exosurf, Pumactant, rabbit natural surfactant from bronchoalveolar lavage, and two synthetic surfactants based on recombinant surfactant protein-C (Venticute) or a leucine/lysine polypeptide. Minimum surface tension, determined with a pulsating bubble surfactometer, was increased >10 mN/m at meconium concentrations >or=0.04 mg/mL for Curosurf, Alveofact, or Survanta, >or=0.32 mg/mL for recombinant surfactant protein-C, >or=1.25 mg/mL for leucine/lysine polypeptide, and >or=20 mg/mL for rabbit natural surfactant. The protein-free synthetic surfactants Exosurf and Pumactant did not reach minimum surface tension <10 mN/m even in the absence of meconium. We conclude that surfactant activity is inhibited by meconium in a dose-dependent manner. Recombinant surfactant protein-C and leucine/lysine polypeptide surfactant were more resistant to inhibition than the modified natural surfactants Curosurf, Alveofact, or Survanta but less resistant than natural lavage surfactant containing surfactant protein-A. We speculate that recombinant hydrophobic surfactant proteins or synthetic analogs of these proteins can be used for the design of new surfactant preparations that are relatively resistant to inactivation and therefore suitable for treatment of acute respiratory distress syndrome. PMID- 11420418 TI - Progressive familial intrahepatic cholestasis with high gamma glutamyltranspeptidase levels in Taiwanese infants: role of MDR3 gene defect? AB - MDR3 P-glycoprotein mediates canalicular phospholipid transport in hepatocytes. Defects in the MDR3 gene have been found to cause a subtype of progressive familial intrahepatic cholestasis (PFIC) with high gamma-glutamyltranspeptidase (GGT) levels. Affected children develop proliferation of biliary epithelium, portal inflammation, and biliary cirrhosis. The frequency of MDR3 mutations in patients with high GGT-PFIC is unclear. There have been no Asian patients reported to carry MDR3 mutations. To determine the role of MDR3 defects in chronic cholestatic patients, we studied six Taiwanese children from five families who presented high GGT-PFIC among 47 patients with infantile onset chronic intrahepatic cholestasis. Sequence analysis of MDR3 cDNA from liver tissues was performed. Only one patient had mutation in the MDR3 gene. This patient had a homozygous 719-bp deletion (nucleotide 287 to 1005) of liver cDNA encompassing exon 5 to 9 and leading to protein truncation. The onset age was 1 y in contrast with the other five patients who presented neonatal cholestasis. Four patients without mutation, including one sibling pair, exhibited histologic features of prominent portal fibrosis leading to advanced biliary cirrhosis that were indistinguishable from the case of MDR3 mutation. We concluded that mutations in MDR3 accounted for approximately 2% (1/47) of infantile onset chronic cholestasis in Taiwan. Those patients presenting high GGT-PFIC with early onset cholestasis but without MDR3 mutation probably had inheritable disorders remaining to be clarified. PMID- 11420419 TI - Reduced Na(+), K(+)-ATPase activity in erythrocyte membranes from patients with phenylketonuria. AB - Na(+), K(+)-ATPase activity was determined in erythrocyte membranes from 12 phenylketonuric patients of both sexes, aged 8.8 +/- 5.0 y, with plasma phenylalanine levels of 0.64 +/- 0.31 mM. The in vitro effects of phenylalanine and alanine on the enzyme activity in erythrocyte membranes from healthy individuals were also investigated. We observed that Na(+), K(+)-ATPase activity was decreased by 31% in erythrocytes from phenylketonuric patients compared with normal age-matched individuals (p < 0.01). We also observed a significant negative correlation between erythrocyte Na(+), K(+)-ATPase activity and plasma phenylalanine levels (r = -0.65; p < 0.05). All PKU patients with plasma phenylalanine levels higher than 0.3 mM had erythrocyte Na(+), K(+)-ATPase activity below the normal range. Phenylalanine inhibited in vitro erythrocyte Na(+), K(+)-ATPase activity by 22 to 34%, whereas alanine had no effect on this activity. However, when combined with phenylalanine, alanine prevented Na(+) K(+) ATPase inhibition. Considering that reduction of Na(+), K(+)-ATPase activity occurs in various neurodegenerative disorders leading to neuronal loss, our previous observations showing a significant reduction of Na(+), K(+)-ATPase activity in brain cortex of rats subjected to experimental phenylketonuria and the present results, it is proposed that determination of Na(+), K(+)-ATPase activity in erythrocytes may be a useful peripheral marker for the neurotoxic effect of phenylalanine in phenylketonuria. PMID- 11420420 TI - Multiple acyl-CoA-dehydrogenase deficiency (MADD): use of acylcarnitines and fatty acids to monitor the response to dietary treatment. AB - The treatment of multiple acyl-CoA-dehydrogenase deficiency (MADD) includes a low fat, low-protein, high-carbohydrate diet, avoiding long fasting periods. However, there is no useful biochemical marker to determine the response to different diets or fasting periods. The aims of this study are to report a patient with MADD, diagnosed through a newborn screening program using tandem mass spectrometry, to assess her response to different feedings, and to evaluate the usefulness of acylcarnitines and FFA to monitor the response to dietary changes. The patient was diagnosed at 6 d. Family history revealed three dead siblings. Five tests were performed, one with breast milk and the subsequent four after giving the patient a bottle of a low-fat, low-protein formula (F), F with glucose polymers (GP), F+GP plus uncooked corn starch (CS), or F+GP+CS preceded by amylase. The results showed that acylcarnitines, FFA, and total nonesterified fatty acids levels were greatly improved at 2 and 4 h on F+GP compared with breast milk. At 6 mo of age, the test with F+CS was repeated to assess the response to a longer fast. The results were similar at 2 and 4 h, but showed a marked increase of acylcarnitines, FFA, and total nonesterified fatty acids at 6 h. The increase of these metabolites could not be avoided by the use of F+GP+CS, but was prevented when amylase was used simultaneously. The patient is currently 3.9 y old and has normal growth and development. We conclude that diagnosis of MADD through a newborn screening program using tandem mass spectrometry is suitable; acylcarnitines and FFA are useful to monitor the response to treatment; and exogenous amylase allows the use of CS in small children with MADD. This therapeutic approach may be an alternative to the use of continuous overnight feedings used for young children with severe fatty acid oxidation defects. Early diagnosis and treatment may change the natural history of MADD. PMID- 11420421 TI - TGF-beta1 regulates the expression of multiple max-interacting transcription factors in Balb/MK cells: implications for understanding the mechanism of action of TGF-beta1. AB - Appropriate transforming growth factor-beta1 (TGF-beta1) signaling is required to preserve homeostasis of diverse tissues during development. At the cellular level, one function of TGF-beta1 that is critical for preserving homeostasis is the ability to arrest cell growth. TGF-beta1 arrests growth by blocking the function of the c-myc proto-oncogene. c-myc function is determined by the level of c-myc expression relative to other Max-interacting transcription factors, and TGF-beta1 has been shown to inhibit c-myc expression by inhibiting c-myc transcription. However, whether TGF-beta1 might also increase the expression of a Max-interacting factor that blocks myc function by competing with myc for Max binding is not known. Therefore, we determined the effect of TGF-beta1 on the expression of Max-interacting transcription factors in Balb/MK cells. We found unexpectedly that Balb/MK cells express both N-myc and c-myc. The pattern of N myc expression during the cell cycle differs from that of c-myc, indicating that mRNA accumulation is controlled by mechanisms specific to each gene. TGF-beta1 rapidly inhibits N-myc mRNA expression; thus N-myc is a novel target of TGF-beta1 in Balb/MK cells. More importantly, we found that TGF-beta1 induces the expression of the putative tumor suppressor genes Mad4 and Mxi1 in both the Balb/MK and Mv1Lu cell lines. Mad4 and Mxi1 are novel targets of TGF-beta1, known to inhibit cell growth by antagonizing the interaction of Myc with Max. Thus, our results suggest that the induction of Mad4 and Mxi1 may function in tandem with the inhibition of N-myc and c-myc to mediate the growth inhibitory function of TGF-beta1. PMID- 11420422 TI - Contribution of reactive oxygen species to 3-hydroxyglutarate neurotoxicity in primary neuronal cultures from chick embryo telencephalons. AB - Glutaryl-CoA dehydrogenase deficiency is an autosomal recessively inherited neurometabolic disorder with a distinct neuropathology characterized by acute encephalopathic crises during a vulnerable period of brain development. 3 Hydroxyglutarate (3-OH-GA), which accumulates in affected patients, has been identified as an endogenous neurotoxin mediating excitotoxicity via N-methyl-D aspartate receptors. As increased generation of reactive oxygen species (ROS) and nitric oxide (NO) plays an important role in excitotoxic neuronal damage, we investigated whether ROS and NO contribute to 3-OH-GA neurotoxicity. 3-OH-GA increased mitochondrial ROS generation in primary neuronal cultures from chick embryo telencephalons, which could be prevented by MK-801, confirming the central role of N-methyl-D-aspartate receptor stimulation in 3-OH-GA toxicity. ROS increase was reduced by alpha-tocopherol and--less effectively-by melatonin. alpha-Tocopherol revealed a wider time frame for neuroprotection than melatonin. Creatine also reduced neuronal damage and ROS formation but only if it was administered >or=6 h before 3-OH-GA. NO production revealed only a slight increase after 3-OH-GA incubation. NO synthase inhibitor N(omega)-nitro-L arginine prevented NO increase but did not protect neurons against 3-OH-GA. The NO donor S-nitroso-N-acetylpenicillamine revealed no effect on 3-OH-GA toxicity at low concentrations (0.5-5 microM), whereas it potentiated neuronal damage at high concentrations (50-500 microM), suggesting that weak endogenous NO production elicited by 3-OH-GA did not affect neuronal viability. We conclude from our results that ROS generation contributes to 3-OH-GA neurotoxicity in vitro and that radical scavenging and stabilization of brain energy metabolism by creatine are hopeful new strategies in glutaryl-CoA dehydrogenase deficiency. PMID- 11420423 TI - Altered expression and function of mitochondrial beta-oxidation enzymes in juvenile intrauterine-growth-retarded rat skeletal muscle. AB - Uteroplacental insufficiency and subsequent intrauterine growth retardation (IUGR) affects postnatal metabolism. In juvenile rats, IUGR alters skeletal muscle mitochondrial gene expression and reduces mitochondrial NAD(+)/NADH ratios, both of which affect beta-oxidation flux. We therefore hypothesized that gene expression and function of mitochondrial beta-oxidation enzymes would be altered in juvenile IUGR skeletal muscle. To test this hypothesis, mRNA levels of five key mitochondrial enzymes (carnitine palmitoyltransferase I, trifunctional protein of beta-oxidation, uncoupling protein-3, isocitrate dehydrogenase, and mitochondrial malate dehydrogenase) and intramuscular triglycerides were quantified in 21-d-old (preweaning) IUGR and control rat skeletal muscle. In isolated skeletal muscle mitochondria, enzyme function of the trifunctional protein of beta-oxidation and isocitrate dehydrogenase were measured because both enzymes compete for mitochondrial NAD(+). Carnitine palmitoyltransferase I, the trifunctional protein of beta-oxidation, and uncoupling protein 3 mRNA levels were significantly increased in IUGR skeletal muscle, whereas mRNA levels of isocitrate dehydrogenase and mitochondrial malate dehydrogenase were unchanged. Similarly, trifunctional protein of beta-oxidation activity was increased in IUGR skeletal muscle mitochondria, and isocitrate dehydrogenase activity was unchanged. Interestingly, skeletal muscle triglycerides were significantly increased in IUGR skeletal muscle. We conclude that uteroplacental insufficiency alters IUGR skeletal muscle mitochondrial lipid metabolism, and we speculate that the changes observed in this study play a role in the long-term morbidity associated with IUGR. PMID- 11420424 TI - Intellectual and psychological performance in males born small for gestational age with and without catch-up growth. AB - Infants born small for gestational age (SGA) have an increased risk of neurologic and intellectual dysfunction. Most of these infants catch up in growth and attain normal height, although some do not. Whether catch-up growth influences intellectual function is not known. To analyze whether intellectual and psychological performance of males in early adulthood are associated with body size at birth or by catch-up growth in height among boys, a population-based cohort was studied. This cohort included all male singletons born without congenital malformations in Sweden from 1973 to 1978 and alive at 18 y (n = 276,033). Information from the Swedish Birth Register was individually linked to the Swedish Conscript Register. Of 254,426 conscripted males, information on intellectual and psychological performance was available for 97% and 91%, respectively. Low birth weight, short birth length, small head circumference at birth, and preterm birth increased the risk of subnormal intellectual and psychological performance. Among SGA-born males, the most important predictor was the absence of catch-up growth. Being born SGA is associated with increased risk of subnormal intellectual and psychological performance. The data strongly support the view that, for males born SGA, it is an advantage to have catch-up growth in length. PMID- 11420425 TI - Inappropriate early hypotension in adolescents: a form of chronic orthostatic intolerance with defective dependent vasoconstriction. AB - Instantaneous orthostatic hypotension (INOH) has been reported in children and adolescents as a new entity of orthostatic intolerance in children who underwent rapid standing as an orthostatic stress test. Children with INOH were discovered among patients presenting with symptoms of chronic orthostatic intolerance, which is often related to orthostatic tachycardia. We used head-up tilt table testing at 70 degrees to investigate children presenting with symptoms of chronic orthostatic intolerance. We compared 24 patients aged 12-17 y, with chronic orthostatic intolerance and symptoms for >or=3 mo, with 13 healthy normal control patients. We recorded continuous heart rate, blood pressure, and respiratory rate and used venous occlusion strain gauge plethysmography to measure calf and forearm blood flow while supine and calf blood flow during head-up tilt. Patients with chronic orthostatic intolerance fulfilled criteria for the postural orthostatic tachycardia syndrome. Postural orthostatic tachycardia syndrome patients were divided into two groups by the occurrence of INOH. Supine forearm and calf arterial resistance was decreased in patients with INOH (n = 8) compared with postural orthostatic tachycardia syndrome patients without INOH (n = 16) and compared with control (n = 13). Resting calf venous pressure was elevated, suggesting excess venous filling because of vasodilation. During early head-up tilt, calf blood flow increased markedly in INOH, less in No-INOH, postural orthostatic tachycardia syndrome patients and least in control patients. Flow was temporally related to calf swelling and negatively correlated to hypotension. The data suggest that INOH occurs in patients with chronic orthostatic intolerance and orthostatic tachycardia and is related to rapid caudal blood flow when upright because of a vasoconstrictor defect. PMID- 11420426 TI - Ornithine aminotransferase messenger RNA expression and enzymatic activity in fetal porcine intestine. AB - In most neonatal animals, the small intestinal epithelium is responsible for endogenous arginine production. The ability of neonatal enterocytes to synthesize arginine immediately after birth suggests that the enzymes involved are present prenatally. Pyrroline-5-carboxylate is the common intermediate in the intestinal pathways for the synthesis of citrulline and arginine from both glutamine and proline and is interconverted into ornithine by ornithine aminotransferase (OAT). In this study, OAT enzymatic activity and mRNA expression in the intestine of fetal pigs from 30 to 110 d of gestation were determined. Enzymatic activity (nanomoles per minute per milligram of protein) peaked at d 45 of gestation and increased again between d 60 and 110 of gestation. At 30 and 35 d of gestation, OAT mRNA expression was detected throughout the mucosal epithelium of the small intestine. Throughout the remainder of gestation, OAT expression was notably higher in the villus epithelium than in the crypt epithelium. The presence of OAT in the small intestinal epithelium throughout gestation suggests that the porcine small intestine is capable of interconverting ornithine and pyrroline-5 carboxylate during fetal development. This capability may be important for synthesis of arginine, proline, ornithine, and polyamines for development and metabolic activity of the intestine during gestation or for somatic growth of the fetus. PMID- 11420427 TI - Apoptosis in various organs of preterm infants: histopathologic study of lung, kidney, liver, and brain of ventilated infants. AB - Apoptosis, the well-characterized form of active programmed cell death, is a physiologic phenomenon in embryonal and fetal life in developing organs. Severe hypoxia, which occurs in most preterm infants, also leads to cell death, which may be necrotic or apoptotic. The aim of our study was to examine the incidence of apoptosis in various organs (such as lung, kidney, and brain) of preterm infants who suffered from clinically proven respiratory distress causing infantile respiratory distress syndrome (IRDS), cardiac failure, and periventricular leukomalacia (PVL). Twenty-four autopsy cases were studied histologically to detect the apoptotic ratio, which was performed on the basis of hematoxylin and eosin staining and validated by terminal deoxynucleotidyl transferase-mediated nick end-labeling (TUNEL) reaction. Elevated apoptotic ratio was found in stages II, III, and IV of bronchopulmonary dysplasia (BPD) among alveolar and bronchiolar cells. The apoptotic activity was very low in stage I of BPD. High apoptotic ratio was detected in hypoxic injuries of the central nervous system (CNS) of preterm infants. Features of apoptosis were present in proximal and excreting tubules of the kidney. Significant elevation of apoptotic activity may play a role in the development of BPD, ischemic brain lesions, and renal failure. PMID- 11420428 TI - Glucose production, gluconeogenesis, and insulin sensitivity in children and adolescents: an evaluation of their reproducibility. AB - The prevalence of overweight and obese children has doubled, and the incidence of type 2 diabetes in children (0-19 y) has increased 4-fold during the past several decades. As a result we can anticipate an increased number of metabolic studies in children. There are few data on measures of glucose metabolism in normal children, and virtually none relating to their reproducibility. The aims of this study were 1) to provide new data on energy expenditure and glucose, lipid, and protein metabolism in nonobese, healthy children and adolescents; 2) to evaluate their reproducibility; and 3) on the basis of these data, to perform power calculations for metabolic studies. Eight nonobese subjects (8-16 y) were studied on two occasions, preceded by 7 d of a diet with identical energy content and macronutrient distribution. Gluconeogenesis, measured by deuterium oxide, accounted for 50% of glucose production. Insulin sensitivity, measured by the labeled minimal model, averaged 4.9 x 10(-4) mL(mU x min)(-1). Glucose appearance rate was significantly higher (p < 0.01) in the children than in the adolescents. Furthermore, we demonstrated that for energy intake and expenditure, plasma concentrations of glucose and C-peptide, and rates of appearance of glucose and leucine, a 10% difference can be detected in fewer than five subjects with a power of 80% and a type I error of 5%. Insulin concentration, gluconeogenesis, insulin secretory indices, insulin sensitivity, and glucose effectiveness were more variable, but with the above power a difference of 25% could be detected in 7-11 subjects using a paired study design. PMID- 11420429 TI - A reliable method for measurement and normalization of pediatric hippocampal volumes. AB - A robust standardized method for segmentation, quantification, and normalization of pediatric hippocampal volumes using magnetic resonance imaging is presented. The method will find application in time course measurements of hippocampal volumes in pediatric patients who suffer from temporal lobe epilepsy and was tested prospectively on six control patients (13-60 mo of age). The un-normalized hippocampal volumes obtained using our segmentation method ranged from 3.85 to 6.38 mL, in agreement with previously published results. Inter- and intraobserver variability of the segmentation method was determined to be 13.3% and 2.8%, respectively. Four different methods of volume normalization were tested. Normalization is required to adjust for age-related increases in hippocampal volume. The normalization approach that seemed to compensate best for growth related hippocampal volume changes was based on a simple estimation of intracranial volumes. This is the first report of a consistent and reliable method for segmentation and normalization of hippocampi from pediatric patients that can be used to study the progression of neurologic diseases in children. PMID- 11420430 TI - Proteins with beta-(thienopyrrolyl)alanines as alternative chromophores and pharmaceutically active amino acids. AB - L-beta-(Thieno[3,2-b]pyrrolyl)alanine and L-beta-(thieno[2,3-b]pyrrolyl)alanine are mutually isosteric and pharmaceutically active amino acids that mimic tryptophan with the benzene ring in the indole moiety replaced by thiophene. Sulfur as a heteroatom causes physicochemical changes in these tryptophan surrogates that bring about completely new properties not found in the indole moiety. These synthetic amino acids were incorporated into recombinant proteins in response to the Trp UGG codons by fermentation in a Trp-auxotrophic Escherichia coli host strain using the selective pressure incorporation method. Related protein mutants expectedly retain the secondary structure of the native proteins but show significantly changed optical and thermodynamic properties. In this way, new spectral windows, fluorescence, polarity, thermodynamics, or pharmacological properties are inserted into proteins. Such an engineering approach by translational integration of synthetic amino acids with a priori defined properties, as shown in this study, proved to be a novel and useful tool for protein rational design. PMID- 11420431 TI - Quantitative evaluation of the lengths of homobifunctional protein cross-linking reagents used as molecular rulers. AB - Homobifunctional chemical cross-linking reagents are important tools for functional and structural characterization of proteins. Accurate measures of the lengths of these molecules currently are not available, despite their widespread use. Stochastic dynamics calculations now provide quantitative measures of the lengths, and length dispersions, of 32 widely used molecular rulers. Significant differences from published data have been found. SUPPLEMENTAL MATERIAL: See www.proteinscience.org PMID- 11420432 TI - Effect of the N2 residue on the stability of the alpha-helix for all 20 amino acids. AB - N2 is the second position in the alpha-helix. All 20 amino acids were placed in the N2 position of a synthetic helical peptide (CH(3)CO-[AXAAAAKAAAAKAAGY]-NH(2)) and the helix content was measured by circular dichroism spectroscopy at 273K. The dependence of peptide helicity on N2 residue identity has been used to determine a free-energy scale by analysis with a modified Lifson-Roig helix-coil theory that includes a parameter for the N2 energy (n2). The rank order of DeltaDeltaG((relative to Ala)) is Glu(-), Asp(-) > Ala > Glu(0), Leu, Val, Gln, Thr, Ile, Ser, Met, Asp(0), His(0), Arg, Cys, Lys, Phe > Asn, > Gly, His(+), Pro, Tyr. The results correlate very well with N2 propensities in proteins, moderately well with N1 and helix interior preferences, and not at all with N-cap preferences. The strongest energetic effects result from interactions with the helix dipole, which favors negative charges at the helix N terminus. Hydrogen bonds to side chains at N2, such as Gln, Ser, and Thr, are weak, despite occurring frequently in protein crystal structures, in contrast to the N-cap position. This is because N-cap hydrogen bonds are close to linear, whereas N2 hydrogen bonds have poor geometry. These results can be used to modify protein stability rationally, help design helices, and improve prediction of helix location and stability. PMID- 11420433 TI - Conformational characterization of oligomeric intermediates and aggregates in beta-lactoglobulin heat aggregation. AB - In one of the first studies of isolated intermediates in protein aggregation, we have used circular dichroism and fluorescence spectroscopy to characterize metastable oligomers that are formed in the early steps of beta-lactoglobulin heat aggregation. The intermediates show typical molten globule characteristics (secondary structure content similar to the native and less tight packing of the side chains), in agreement with the belief that partly folded states play a key role in protein aggregation. The far-UV CD signal bears strong resemblance to that of a known folding intermediate. Cryo-transmission electron microscopy of the aggregates reveals spherical particles with a diameter of about 50 nm and an internal threadlike structure. Isolated oligomers as well as larger aggregates bind the dye thioflavin T, usually a signature of the amyloid superstructures found in many protein aggregates. This result suggests that the structural motif recognized by thioflavin T can be formed in small oligomers. PMID- 11420434 TI - Correlated conformational fluctuations during enzymatic catalysis: Implications for catalytic rate enhancement. AB - Correlated enzymatic conformational fluctuations are shown to contribute to the rate of enhancement achieved during catalysis. Cytidine deaminase serves as a model system. Crystallographic temperature factor data for this enzyme complexed with substrate analog, transition-state analog, and product are available, thereby establishing a measure of atomic scale conformational fluctuations along the (approximate) reaction coordinate. First, a neural network-based algorithm is used to visualize the decreased conformational fluctuations at the transition state. Second, a dynamic diffusion equation along the reaction coordinate is solved and shows that the flux velocity through the associated enzymatic conformation space is greatest at the transition state. These results suggest (1) that there are both dynamic and energetic restrictions to conformational fluctuations at the transition state, (2) that enzymatic catalysis occurs on a fluctuating potential energy surface, and (3) a form for the potential energy. The Michaelis-Menten equations are modified to describe catalysis on this fluctuating potential energy profile, leading to enhanced catalytic rates when fluctuations along the reaction coordinate are appropriately correlated. This represents a dynamic tuning of the enzyme for maximally effective transformation of the ES complex into EP. PMID- 11420435 TI - The energetic cost of induced fit catalysis: Crystal structures of trypsinogen mutants with enhanced activity and inhibitor affinity. AB - The contribution of induced fit to enzyme specificity has been much debated, although with little experimental data. Here we probe the effect of induced fit on enzyme specificity using the trypsin(ogen) system. BPTI is known to induce trypsinogen to assume a trypsinlike conformation. Correlations are observed between BPTI affinity and the values of k(cat)/K(m) for the hydrolysis of two substrates by eight trypsin(ogen) variants. The slope of both correlations is 1.8. The crystal structures of the BPTI complexes of four variant trypsinogens were also solved. Three of these enzymes, K15A, DeltaI16V17/D194N, and DeltaI16V17/Q156K trypsinogen, are 10- to 100-fold more active than trypsinogen. The fourth variant, DeltaI16V17 trypsinogen, is the lone outlier in the correlations; its activity is lower than expected based on its affinity for BPTI. The S1 site and oxyanion hole, formed by segments 184A-194 and 216-223, are trypsinlike in all of the enzymes. These structural and kinetic data confirm that BPTI induces an active conformation in the trypsin(ogen) variants. Thus, changes in BPTI affinity monitor changes in the energetic cost of inducing a trypsinlike conformation. Although the S1 site and oxyanion hole are similar in all four variants, the N-terminal and autolysis loop (residues 142-152) segments have different interactions for each variant. These results indicate that zymogen activity is controlled by a simple conformational equilibrium between active and inactive conformations, and that the autolysis loop and N-terminal segments control this equilibrium. Together, these data illustrate that induced fit does not generally contribute to enzyme specificity. PMID- 11420436 TI - Heat capacity changes upon burial of polar and nonpolar groups in proteins. AB - In this paper we address the question of whether the burial of polar and nonpolar groups in the protein locale is indeed accompanied by the heat capacity changes, DeltaC(p), that have an opposite sign, negative for nonpolar groups and positive for polar groups. To accomplish this, we introduced amino acid substitutions at four fully buried positions of the ubiquitin molecule (Val5, Val17, Leu67, and Gln41). We substituted Val at positions 5 and 17 and Leu at position 67 with a polar residue, Asn. As a control, Ala was introduced at the same three positions. We also replaced the buried polar Gln41 with Val and Leu, nonpolar residues that have similar size and shape as Gln. As a control, Asn was introduced at Gln41 as well. The effects of these amino acid substitutions on the stability, and in particular, on the heat capacity change upon unfolding were measured using differential scanning calorimetry. The effect of the amino acid substitutions on the structure was also evaluated by comparing the (1)H-(15)N HSQC spectra of the ubiquitin variants. It was found that the Ala substitutions did not have a considerable effect on the heat capacity change upon unfolding. However, the substitutions of aliphatic side chains (Val or Leu) with a polar residue (Asn) lead to a significant (> 30%) decrease in the heat capacity change upon unfolding. The decrease in heat capacity changes does not appear to be the result of significant structural perturbations as seen from the HSQC spectra of the variants. The substitution of a buried polar residue (Gln41) to a nonpolar residue (Leu or Val) leads to a significant (> 25%) increase in heat capacity change upon unfolding. These results indicate that indeed the heat capacity change of burial of polar and nonpolar groups has an opposite sign. However, the observed changes in DeltaC(p) are several times larger than those predicted, based on the changes in water accessible surface area upon substitution. PMID- 11420437 TI - Aberrant mobility phenomena of the DNA repair protein XPA. AB - The DNA repair protein XPA recognizes a wide variety of bulky lesions and interacts with several other proteins during nucleotide excision repair. We recently identified regions of intrinsic order and disorder in full length Xenopus XPA (xXPA) protein using an experimental approach that combined time resolved trypsin proteolysis and electrospray ionization interface coupled to a Fourier transform ion cyclotron resonance (ESI-FTICR) mass spectrometry (MS). MS data were consistent with the interpretation that xXPA contains no post translational modifications. Here we characterize the discrepancy between the calculated molecular weight (31 kDa) for xXPA and its apparent molecular weight on SDS-PAGE (multiple bands from approximately 40-45 kDa) and gel filtration chromatography ( approximately 92 kDa), as well as the consequences of DNA binding on its anomalous mobility. Iodoacetamide treatment of xXPA prior to SDS PAGE yielded a single 42-kDa band, showing that covalent modification of Cys did not correct aberrant mobility. Determination of sulfhydryl content in xXPA with Ellman's reagent revealed that all nine Cys in active protein are reduced. Unexpectedly, structural constraints induced by intramolecular glutaraldehyde crosslinks in xXPA produced a approximately 32-kDa monomer in closer agreement with its calculated molecular weight. To investigate whether binding to DNA alters xXPA's anomalous migration, we used gel filtration chromatography. For the first time, we purified stable complexes of xXPA and DNA +/- cisplatin +/- mismatches. xXPA showed at least 10-fold higher affinity for cisplatin DNA +/- mismatches compared to undamaged DNA +/- mismatches. In all cases, DNA binding did not correct xXPA's anomalous migration. To test predictions that a Glu-rich region (EEEEAEE) and/or disordered N- and C-terminal domains were responsible for xXPA's aberrant mobility, the molecular weights of partial proteolytic fragments from approximately 5 to 25 kDa separated by reverse-phase HPLC and precisely determined by ESI-FTICR MS were correlated with their migration on SDS-PAGE. Every partial tryptic fragment analyzed within this size range exhibited 10%-50% larger molecular weights than expected. Thus, both the disordered domains and the Glu-rich region in xXPA are primarily responsible for the aberrant mobility phenomena. PMID- 11420438 TI - Ligand-modulation of the stability of the glucose transporter GLUT 1. AB - The glucose transporter GLUT 1 was isolated from human erythrocytes and reconstituted into endogenous membrane lipids. Results from thermal denaturation studies, using differential scanning calorimetry, indicate that the thermal denaturation temperature of GLUT 1 is significantly lower in the presence of ATP. The lowering of this transition temperature is very dependent on pH. At more acidic pH, ATP has a greater effect of lowering the thermal denaturation temperature of the protein. For example, with 4.8 mM ATP, the denaturation endotherm is lowered by over 10 degrees at pH 4.3, whereas at pH 7.4, ATP does not alter this transition temperature. However, a change in pH alone, in the absence of ATP, has very little effect on the denaturation temperature. Both glucose and salt partially reverse the lowering of the temperature of thermal denaturation caused by ATP. Studies of acrylamide quenching of the Trp residues of GLUT 1 indicate that at neutral pH, ATP increases the Stern-Volmer quenching constant, while glucose lowers it. The results indicate that ATP binds to GLUT 1 and destabilizes the native structure, leading to a lowering of the thermal denaturation temperature and an increase in acrylamide quenching. The effects of ATP are reversed in part by glucose and are also partly electrostatic in nature. PMID- 11420439 TI - Mass spectrometric study of the Escherichia coli repressor proteins, Ic1R and Gc1R, and their complexes with DNA. AB - In Escherichia coli, the IclR protein regulates both the aceBAK operon and its own synthesis. Database homology searches have identified many IclR-like proteins, now known as the IclR family, which can be identified by a conserved C terminal region. We have cloned and purified one of these proteins, which we have named GclR (glyoxylate carboligase repressor). Although purification is straightforward, both the IclR and GclR proteins are difficult to manipulate, requiring high salt (up to 0.6 M KCl) for solubility. With the advent of nanospray ionization, we could transfer the proteins into much higher concentrations of volatile buffer than had been practical with ordinary electrospray. In 0.5 M ammonium bicarbonate buffer, both proteins were stable as tetramers, with a small amount of dimer. In a separate experiment, we found that IclR protein selected from a random pool a sequence which matched exactly that of the presumed binding region of the GclR protein, although IclR does not regulate the gcl gene. We designed a 29 bp synthetic DNA to which IclR and GclR bind, and with which we were able to form noncovalent DNA-protein complexes for further mass spectrometry analysis. These complexes were far more stable than the proteins alone, and we have evidence of a stoichiometry which has not been described previously with (protein monomer : dsDNA) = (4 : 1). PMID- 11420440 TI - Elongation of the BH8 beta-hairpin peptide: Electrostatic interactions in beta hairpin formation and stability. AB - An elongated version of the de novo designed beta-hairpin peptide, BH8, has allowed us to gain insight into the role of electrostatic interactions in beta hairpin stability. A Lys-Glu electrostatic pair has been introduced by adding a residue at the beginning and at the end of the N-terminal and C-terminal strands, respectively, of the beta-hairpin structure, in both orientations. The two resulting peptides and controls having Ala residues at these positions and different combinations of Ala with Lys, or Glu residues, have been analyzed by nuclear magnetic resonance (NMR), under different pH and ionic strength conditions. All of the NMR parameters, in particular the conformational shift analysis of Calpha protons and the coupling constants, (3)J(HNalpha), correlate well and the population estimates are in reasonable agreement among the different methods used. In the most structured peptides, we find an extension of the beta hairpin structure comprising the two extra residues. Analysis of the pH and salt dependence shows that ionic pairs contribute to beta-hairpin stability. The interaction is electrostatic in nature and can be screened by salt. There is also an important salt-independent contribution of negatively charged groups to the stability of this family of beta-hairpin peptides. PMID- 11420441 TI - Backbone dynamics of the regulatory domain of calcium vector protein, studied by (15)N relaxation at four fields, reveals unique mobility characteristics of the intermotif linker. AB - CaVP is a calcium-binding protein from amphioxus. It has a modular composition with two domains, but only the two EF-hand motifs localized in the C-terminal domain are functional. We recently determined the solution structure of this regulatory half (C-CaVP) in the Ca(2+)-saturated form and characterized the stepwise ion binding. This paper reports the (15)N nuclear relaxation rates of the Ca(2+)-saturated C-CaVP, measured at four different NMR fields (9.39, 11.74, 14.1, and 18.7 T), which were used to map the spectral density function for the majority of the amide H(N)-N vectors. Fitting the spectral density values at eight frequencies by a model-free approach, we obtained the microdynamic parameters characterizing the global and internal movements of the polypeptide backbone. The two EF-hand motifs, including the ion binding loops, behave like compact structural units with restricted mobility as reflected in the quite uniform order parameter and short internal correlation time (< 20 nsec). Comparative analysis of the two Ca(2+) binding sites shows that site III, having a larger affinity for the metal ion, is generally more rigid, and the amide vector in the second residue of each loop is significantly less restricted. The linker fragment is animated simultaneously by a larger amplitude fast motion and a slow conformational exchange on a microsecond to millisecond time scale. The backbone dynamics of C-CaVP characterized here is discussed in relation with other well-characterized Ca(2+)-binding proteins. SUPPLEMENTAL MATERIAL: See www.proteinscience.org PMID- 11420442 TI - Enzyme specificity under dynamic control II: Principal component analysis of alpha-lytic protease using global and local solvent boundary conditions. AB - The contributions of conformational dynamics to substrate specificity have been examined by the application of principal component analysis to molecular dynamics trajectories of alpha-lytic protease. The wild-type alpha-lytic protease is highly specific for substrates with small hydrophobic side chains at the specificity pocket, while the Met190-->Ala binding pocket mutant has a much broader specificity, actively hydrolyzing substrates ranging from Ala to Phe. Based on a combination of multiconformation analysis of cryo-X-ray crystallographic data, solution nuclear magnetic resonance (NMR), and normal mode calculations, we had hypothesized that the large alteration in specificity of the mutant enzyme is mainly attributable to changes in the dynamic movement of the two walls of the specificity pocket. To test this hypothesis, we performed a principal component analysis using 1-nanosecond molecular dynamics simulations using either a global or local solvent boundary condition. The results of this analysis strongly support our hypothesis and verify the results previously obtained by in vacuo normal mode analysis. We found that the walls of the wild type substrate binding pocket move in tandem with one another, causing the pocket size to remain fixed so that only small substrates are recognized. In contrast, the M190A mutant shows uncoupled movement of the binding pocket walls, allowing the pocket to sample both smaller and larger sizes, which appears to be the cause of the observed broad specificity. The results suggest that the protein dynamics of alpha-lytic protease may play a significant role in defining the patterns of substrate specificity. As shown here, concerted local movements within proteins can be efficiently analyzed through a combination of principal component analysis and molecular dynamics trajectories using a local solvent boundary condition to reduce computational time and matrix size. PMID- 11420443 TI - A critical investigation of the Tanford-Kirkwood scheme by means of Monte Carlo simulations. AB - Monte Carlo simulations are used to assess the adequacy of the Tanford-Kirkwood prescription for electrostatic interactions in macromolecules. Within a continuum dielectric framework, the approach accurately describes salt screening of electrostatic interactions for moderately charged systems consistent with common proteins at physiological conditions. The limitations of the Debye-Huckel theory, which forms the statistical mechanical basis for the Tanford-Kirkwood result, become apparent for highly charged systems. It is shown, both by an analysis of the Debye-Huckel theory and by numerical simulations, that the difference in dielectric permittivity between macromolecule and surrounding solvent does not play a significant role for salt effects if the macromolecule is highly charged. By comparison to experimental data, the continuum dielectric model (combined with either an approximate effective Hamiltonian as in the Tanford-Kirkwood treatment or with exact Monte Carlo simulations) satisfactorily predicts the effects of charge mutation on metal ion binding constants, but only if the macromolecule and solvent are assigned the same or similar permittivities. PMID- 11420444 TI - Kinetic properties and inhibition of the dimeric dUTPase-dUDPase from Leishmania major. AB - Kinetic properties of the dimeric enzyme dUTPase from Leishmania major were studied using a continuous spectrophotometric method. dUTP was the natural substrate and dUMP and PPi the products of the hydrolysis. The trypanosomatid enzyme exhibited a low K(m) value for dUTP (2.11 microM), a k(cat) of 49 s(-1), strict Michaelis-Menten kinetics and is a potent catalyst of dUDP hydrolysis, whereas in other dUTPases described, this compound acts as a competitive inhibitor. Discrimination is achieved for the base and sugar moiety showing specificity constants for different dNTPs similar to those of bacterial, viral, and human enzymes. In the alkaline range, the K(m) for dUTP increases with the dissociation of ionizable groups showing pK(a) values of 8.8, identified as the uracil moiety of dUTP and 10, whereas in the acidic range, K(m) is regulated by an enzyme residue exhibiting a pK(a) of 7.1. Activity is strongly inhibited by the nucleoside triphosphate analog alpha-beta-imido-dUTP, indicating that the enzyme can bind triphosphate analogs. The existence of specific inhibition and the apparent structural and kinetic differences (reflected in different binding strength of dNTPs) with other eukaryotic dUTPases suggest that the present enzyme might be exploited as a target for new drugs against leishmaniasis. PMID- 11420445 TI - Experimental determination and calculations of redox potential descriptors of compounds directed against retroviral zinc fingers: Implications for rational drug design. AB - A diverse set of electrophilic compounds that react with cysteine thiolates in retroviral nucleocapsid (NC) proteins and abolish virus infectivity has been identified. Although different in chemical composition, these compounds are all oxidizing agents that lead to the ejection of Zn(II) ions bound to conserved structural motifs (zinc fingers) present in retroviral NC proteins. The reactivity of a congeneric series of aromatic disulfides toward the NC protein of the human immunodeficiency virus type 1 (HIV-1), NCp7, has been characterized by HPLC separation of starting reagents from reaction products. We calculated the absolute redox potentials of these compounds in the gas phase and in aqueous solvent, using a density functional theory method and a continuum solvation model. Pulsed polarography experiments were performed and showed a direct correlation between calculated and experimentally determined redox propensities. A dependence between protein reactivity and redox potential for a specific compound was shown: Reaction with NCp7 did not take place below a threshold value of redox potential. This relationship permits the distinction between active and nonactive compounds targeted against NCp7, and provides a theoretical basis for a scale of reactivity with retroviral zinc fingers. Our results indicate that electrophilic agents with adequate thiophilicity to react with retroviral NC fingers can now be designed using known or calculated electrochemical properties. This may assist in the design of antiretroviral compounds with greater specificity for NC protein. Such electrophilic agents can be used in retrovirus inactivation with the intent of preparing a whole-killed virus vaccine formulation that exhibits unaffected surface antigenic properties. PMID- 11420446 TI - Cavities of alpha(1)-antitrypsin that play structural and functional roles. AB - The native form of inhibitory serine protease inhibitors (serpins) is strained, which is critical for their inhibitory activity. Previous studies on stabilizing mutations of alpha(1)-antitrypsin, a prototype of serpins, indicated that cavities provide a structural basis for the native strain of the molecule. We have systematically mapped the cavities of alpha(1)-antitrypsin that play such structural and functional roles by designing cavity-filling mutations at residues that line the walls of the cavities. Results show that energetically unfavorable cavities are distributed throughout the alpha(1)-antitrypsin molecule, and the cavity-filling mutations stabilized the native conformation at 8 out of 10 target sites. The stabilization effect of the individual cavity-filling mutations of alpha(1)-antitrypsin varied (0.2-1.9 kcal/mol for each additional methylene group) and appeared to depend largely on the structural flexibility of the cavity environment. Cavity-filling mutations that decreased inhibitory activity of alpha(1)-antitrypsin were localized in the loop regions that interact with beta sheet A distal from the reactive center loop. The results are consistent with the notion that beta-sheet A and the structure around it mobilize when alpha(1) antitrypsin forms a complex with a target protease. PMID- 11420447 TI - Increasing protein stability using a rational approach combining sequence homology and structural alignment: Stabilizing the WW domain. AB - This study shows that a combination of sequence homology and structural information can be used to increase the stability of the WW domain by 2.5 kcal mol(-1) and increase the T(m) by 28 degrees C. Previous homology-based protein design efforts typically investigate positions with low sequence identity, whereas this study focuses on semi-conserved core residues and proximal residues, exploring their role(s) in mediating stabilizing interactions on the basis of structural considerations. The A20R and L30Y mutations allow increased hydrophobic interactions because of complimentary surfaces and an electrostatic interaction with a third residue adjacent to the ligand-binding hydrophobic cluster, increasing stability significantly beyond what additivity would predict for the single mutations. The D34T mutation situated in a pi-turn possibly disengages Asn31, allowing it to make up to three hydrogen bonds with the backbone in strand 1 and loop 2. The synergistic mutations A20R/L30Y in combination with the remotely located mutation D34T add together to create a hYap WW domain that is significantly more stable than any of the protein structures on which the design was based (Pin and FBP28 WW domains). PMID- 11420448 TI - Application of high-precision isotope ratio monitoring mass spectrometry to identify the biosynthetic origins of proteins. AB - Isotope ratio monitoring (IRM) mass spectrometry was used to measure the relative abundance of stable isotopes in several samples of adult human hemoglobin expressed in E. coli, yeast, and human blood. The results showed significant differences in the distribution of (15)N and (13)C isotopes among hemoglobin samples produced in these organisms. This indicates that IRM mass spectrometry can be used in forensic protein chemistry to identify the origin of protein expression. PMID- 11420449 TI - A normalized root-mean-square distance for comparing protein three-dimensional structures. AB - The degree of similarity of two protein three-dimensional structures is usually measured with the root-mean-square distance between equivalent atom pairs. Such a similarity measure depends on the dimension of the proteins, that is, on the number of equivalent atom pairs. The present communication presents a simple procedure to make the root-mean-square distances between pairs of three dimensional structures independent of their dimensions. This normalization may be useful in evolutionary and fold classification studies as well as in simple comparisons between different structural models. PMID- 11420450 TI - The wounded healer. AB - This paper deals with the emergence, elaboration, and use of the concept of "the wounded healer." The term refers to a person whose personal experience of illness and/or trauma has left lingering effects on him--in the form of lessons learned that later served him in ministering to other sufferers, or in the form of symptoms or characteristics that usefully influenced his therapeutic endeavors. While such persons and their actions have been noted across the ages, in other cultures, and in many contexts, it was not until the early twentieth century that the patterns in the behaviors of such persons were recognized, named, explained, and categorized as "healing." Early in the century, the concept was commonly used in the fields of pastoral counseling and analytical psychology; by the end of the century it had been vastly expanded and extended and no longer referred mainly to a healer of psychological suffering. The term wounded healer is now in common use in areas such as rehabilitation medicine, medical-career choice, Alcoholics Anonymous and the self-help movement, and chronic-illness support groups, as well as in the original areas of psychotherapy and pastoral care. PMID- 11420451 TI - The origins of informed consent: the International Scientific Commission on Medical War Crimes, and the Nuremburg code. AB - The Nuremberg Code has generally been seen as arising from the Nuremberg Medical Trial. This paper examines developments prior to the Trial, involving the physiologist Andrew Conway Ivy and an inter-Allied Scientific Commission on Medical War Crimes. The paper traces the formulation of the concept of a medical war crime by the physiologist John West Thompson, as part of the background to Ivy's code on human experiments of 1 August 1946. It evaluates subsequent responses by the American Medical Association, and by other war crimes experts, notably Leo Alexander, who developed Ivy's conceptual framework. Ivy's interaction with the judges at Nuremberg alerted them to the importance of formulating ethical guidelines for clinical research. PMID- 11420452 TI - Sex research at the borders of gender: transvestites, transsexuals, and Alfred C. Kinsey. AB - Sex researcher Alfred Kinsey's vision of sexual taxonomy continued to evolve after he published his first landmark volume on human sexuality, and his research into sexual subcultures went beyond his initial studies of homosexuality and prostitution. In the late 1940s and early 1950s, he developed a new interest in cross-dressing and cross-gender identification. This article outlines how and why he began to interview transvestites and transsexuals, and places his emerging vision of gendered behavior and gender identity within the scientific theories of his day. Kinsey rejected the prevailing views, preferring instead a behaviorist model of gender. He saw cross-dressing and crossgender identification as male phenomena and used them to speculate about sex differences in the capacity for psychological conditioning. In his usual style, he did not condemn transvestites or transsexuals, but he disapproved of the genital surgery requested by male-to female transsexuals. It was here that Kinsey hit the limits of his well-known sexual liberalism in which he approved of all sexual variations that did not involve coercion. PMID- 11420453 TI - Lupines, manganese, and devil-sickness: an Anglo-Saxon medical response to epilepsy. AB - The most frequently prescribed herb for "devil-sickness" in the vernacular medical books from Anglo-Saxon England, the lupine, is exceptionally high in manganese. Since manganese depletion has been linked with recurring seizures in both clinical and experimental studies, it is possible that lupine administration responded to the particular pathophysiology of epilepsy. Lupine is not prescribed for seizures in classical Mediterranean medical sources, implying that the Northern European peoples (if not the Anglo-Saxons themselves) discovered whatever anticonvulsive properties the herb may exhibit. PMID- 11420454 TI - Mucoepidermoid carcinoma: a clinicopathologic study of 80 patients with special reference to histological grading. AB - We sought to review our experience with salivary mucoepidermoid carcinoma (MEC) over two decades to confirm the validity and reproducibility of histologic grading and to investigate MIB-1 index as a prognosticator. Diagnosis was confirmed on 80 cases, and chart review or patient contact was achieved for 48 patients, with follow-up from 5 to 240 months (median 36 months). Immunohistochemistry with citrate antigen retrieval for MIB-1 was performed on a subset of cases. Kaplan-Meier survival curves were generated for each stage, site, and grade according to our proposed grading system. To address the issue of grading reproducibility, 20 slides were circulated among five observers, without prior discussion; slides were categorized as low-, intermediate-, or high-grade according to one's "own" criteria, and then according to the AFIP criteria proposed by Goode et al.10 Weighted kappa (kappa) estimates were obtained to describe the extent of agreement between pairs of rating. The Wilcoxon signed rank test or the Friedman test as appropriate tested variation across ratings. There was no gender predominance and a wide age range (15-86 years, median 49 years). The two most common sites were parotid and palate. All grade 1 MECs presented as Stage I tumors, and no failures were seen for this category. The local disease failure rates at 75 months for grades 2 and 3 MEC were 30% and 70%, respectively. Tumor grade, stage, and negative margin status all correlated with disease-free survival (DFS) (p = 0.0091, 0.0002, and 0.048, respectively). The MIB index was not found to be predictive of grade. Regarding the reproducibility of grading, the interobserver variation for pathologists using their "own" grading, as expressed by the kappa value, ranged from good agreement (kappa = 0.79) to poor (kappa = 0.27) (average kappa = 0.49). A somewhat better interobserver reproducibility was achieved when the pathologists utilized the standardized AFIP criteria (average kappa = 0.61, range 0.38-0.77). This greater agreement was also reflected in the Friedman test (statistical testing of intraobserver equality), which indicated significant differences in using one's own grading systems (p = 0.0001) but not in applying the AFIP "standardized" grading (p = 0.33). When one's own grading was compared with the AFIP grading, there were 100 pairs of grading "events," with 46 disagreements/100 pairs. For 98% of disagreements, the AFIP grading "downgraded" tumors. This led us to reanalyze a subset of 31 patients for DFS versus grade, for our grading schema compared with the AFIP grading. Although statistical significance was not achieved for this subset, the log rank value revealed a trend for our grading (p = 0.0993) compared with the Goode schema (p = 0.2493). This clinicopathologic analysis confirms the predictive value of tumor staging and three-tiered histologic grading. Our grading exercise confirms that there is significant grading disparity for MEC, even among experienced ENT/oral pathologists. The improved reproducibility obtained when the weighted AFIP criteria were used speaks to the need for an accepted and easily reproducible system. However, these proposed criteria have a tendency to downgrade MEC. Therefore, the addition of other criteria (such as vascular invasion, pattern of tumor infiltration [i.e., small islands and individual cells vs cohesive islands]) is necessary. We propose a modified grading schema, which enhances predictability and provides much needed reproducibility. PMID- 11420455 TI - Schwannomas in the colon and rectum: a clinicopathologic and immunohistochemical study of 20 cases. AB - Schwannomas of the colon and rectum are uncommon and incompletely characterized tumors, and only a small number of cases have been reported. This study was undertaken to determine the clinicopathologic profile of such tumors. A total of 20 colorectal schwannomas were identified and analyzed in a review of 600 mesenchymal tumors of the colon and rectum from the files of the Armed Forces Institute of Pathology. The schwannomas occurred equally in men (n = 9) and women (n = 11) in a wide age range (18-87 years; median age 65 years). The most common location was cecum (n = 7), followed by sigmoid and rectosigmoid (n = 6), transverse colon (n = 3), descending colon (n = 2), and rectum (n = 1); the location of one tumor had not been specified. The tumors commonly presented as polypoid intraluminal lesions, often with mucosal ulceration. Rectal bleeding, colonic obstruction, and abdominal pain were the most common presenting symptoms. The most common histologic variant (n = 15) was a spindle cell schwannoma with a trabecular pattern and vague or no Verocay bodies. These tumors ranged from 0.5 to 5.5 cm in diameter. A lymphoid cuff with germinal centers typically surrounded these tumors and focal nuclear atypia was often present, but mitotic activity never exceeded 5 per 50 HPF. All four epithelioid schwannomas occurred in the descending colon or sigmoid, three of them as small submucosal tumors. There was one plexiform schwannoma in the sigmoid composed of multiple nodules of prominently palisading schwann cells similar to those seen in conventional soft tissue schwannomas. All tumors studied were strongly positive for S-100 protein and also for low affinity nerve growth factor receptor (p75), collagen IV, and GFAP. Three tumors had CD34-positive cells, but all were negative for CD117 (KIT), neurofilament proteins, smooth muscle actin, and desmin. The percentage of MIB-1-positive cells was usually less than 1% and never higher than 3%. Colorectal schwannomas behaved in a benign fashion with no evidence of aggressive behavior or connection with neurofibromatosis 1 or 2, based on follow-up information on 18 patients. PMID- 11420456 TI - Rhabdomyosarcoma of the urinary bladder and vagina: a clinicopathologic study with emphasis on recurrent disease: a report from the Kiel Pediatric Tumor Registry and the German CWS Study. AB - Rhabdomyosarcomas (RMS) of the urinary bladder and vagina vary in their biologic and clinical behavior and require different types of treatment. Anatomically the two organs are close, and the reason for these differences in behavior is unknown. We investigated tumor specimens of 51 urinary bladder RMS and 14 vaginal RMS with regard to histologic subtype, growth pattern, differentiation, and proliferation morphologically and immunohistochemically. Recurrences and/or "second look" specimens from 15 patients after chemotherapy were compared with the primary tumors. Within the 65 specimens we found 31 "classical" embryonal RMS, 26 embryonal RMS of botryoid subtype (BRMS), 3 embryonal RMS of spindle cell subtype, and 5 alveolar RMS. BRMS is more common in the vagina (11 BRMS of 14 cases) than in the urinary bladder RMS (15 BRMS of 54 cases). Classical embryonal RMS with a polypoid (exophytic) growth pattern is associated with a more favorable prognosis (92% 10-year survival) than the same type with a diffuse intramural (endophytic) growth pattern (68% 10-year survival, p = 0.02). The proliferation rate was associated with the degree of differentiation, but neither showed a correlation with prognosis. A marked maturation after chemotherapy was seen in the majority of recurrences and SL specimens, associated with lowered proliferation activity. Two of 12 patients with recurrences showing chemotherapy induced maturation died of the disease. In conclusion, we determined that polypoid embryonal RMS of both the urinary bladder and the vagina have a comparably good prognosis. This includes all botryoid RMS. The poorer prognosis of the group of urinary bladder RMS as a whole was caused by a high incidence of diffusely growing RMS, which have a less favorable prognosis than polypoid RMS. Maturation after chemotherapy occurs frequently in RMS. In contrast to the excellent prognosis reported in other studies, we had two patients with fatal outcome despite chemotherapy-induced maturation in the recurrences. PMID- 11420457 TI - Bone marrow involvement in lymphomas with hemophagocytic syndrome at presentation: a clinicopathologic study of 11 patients in a Western institution. AB - Hemophagocytic syndrome (HPS) is a clinicopathologic syndrome that can reveal a non-Hodgkin's lymphoma. The pathologic features of lymphoma associated with HPS remain ill defined. We studied 11 lymphomas associated with HPS on initial bone marrow biopsies, consecutively diagnosed during a 6-year period in a Western institution. There were seven diffuse large B-cell lymphomas (DLBCLs), three T cell lymphomas (one peripheral T-cell lymphoma unspecified, two hepatosplenic gammadelta T-cell lymphomas [HS gammadeltaTLs]), and one aggressive NK-cell lymphoma/leukemia (NKL). These lymphomas shared common clinicopathologic features with a systemic presentation, a poor outcome (nine patients died within 2 years), and a mild interstitial lymphoid infiltrate of the bone marrow at presentation in nine patients. This equivocal lymphoma infiltrate was blending with normal hematopoietic cells, and CD20 and CD3 immunolabelings were essential for its detection. A high number of reactive T (CD3+) cells, most often with a predominant cytotoxic (CD8+ TiA1+) phenotype, was present in all DLBCLs. By in situ hybridization, Epstein-Barr virus was detected in neoplastic cells of three cases (one DLBCL, one HS gammadeltaTL, and one NKL), which also showed serum viral DNA. Polymerase chain reaction studies disclosed HHV6 DNA sequences in tumor tissues of two DLBCLs, whereas HHV8 DNA was not detected. Because tumor mass indicative of lymphoma was not striking in most patients, bone marrow biopsy appears to be of great value for the diagnosis of an HPS-associated lymphoma, which may be, in Western patients, of B- as well as T- or NK-cell type. Immunostaining for CD3 and CD20 is essential to identify the common subtle lymphoma involvement. Together with a better understanding of the pathogenic processes, an early diagnosis may improve the prognosis of HPS-associated lymphoma. PMID- 11420458 TI - Cutaneous follicular B-cell lymphoma: description of a series of 18 cases. AB - The lack of precise and homogeneous criteria for the recognition of primary cutaneous follicular lymphoma has hindered gaining data on the frequency and clinical and molecular features of this entity. In the course of a review of a series of primary cutaneous lymphoma from different Spanish hospitals, we collected a series of 18 cases of primary cutaneous follicular lymphoma and analyzed its clinical, morphologic, and biologic characteristics. In this review only cases with a follicular pattern of growth, germinal center cytology, and restriction to the skin in a minimum follow-up of 6 months have been included. Cases of primary cutaneous follicular lymphoma were characterized by the expression of classic markers of the germinal center, such as bcl6, CD10, and the presence of aggregates of follicular dendritic cells. They frequently express bcl2 protein, although classical t(14;18) was not found in any of the cases analyzed. Analysis of the bcl6 noncoding first exon showed somatic mutations in two of four cases analyzed, as would be expected in lymphoma deriving from the germinal center. Clinically, most cases showed initial involvement of the head and neck, with relapses in eight cases (involving the skin in five cases, both skin and lymph node in two cases, and lymph node in one case). No death attributable to the tumor was recorded. These data seem to imply that follicular lymphoma may present initially in the skin, lacking the characteristic t(14;18) and having a relatively indolent course. Recognition of these tumors and elucidation of their molecular alterations could lead to properly adapted staging and treatment protocols for these patients. PMID- 11420459 TI - Ki-67, cyclin E, and p16INK4 are complimentary surrogate biomarkers for human papilloma virus-related cervical neoplasia. AB - Prior studies of Ki-67, cyclin E, and p16 expression have suggested that these biomarkers may be preferentially expressed in cervical neoplasia. This study examined and compared the distribution of staining for these three antigens in 1) normal and reactive epithelial changes, 2) diagnostically challenging cases (atypical metaplasia and atypical atrophy), 3) squamous intraepithelial lesions (SIL), and 4) high-and low-risk human papilloma virus (HPV) type-specific SIL. One hundred four epithelial foci from 99 biopsies were studied, including low grade squamous intraepithelial lesions (LSIL; 24), high-grade squamous intraepithelial lesions (HSIL; 36), mature or immature (metaplastic) squamous epithelium (29), and atrophic or metaplastic epithelium with atypia (15). Cases were scored positive for Ki-67 expression if expression extended above the basal one third of the epithelium, for cyclin E if moderate to strong staining was present, and for p16 if moderate to strong diffuse or focal staining was present. HPV status was scored by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of extracted DNA. Immunohistochemical findings were correlated with histologic and viral data. Overall, a histologic diagnosis of SIL correlated strongly with all of the biomarkers used (p <0.001). Positive scores for Ki-67, cyclin E, and p16 were seen in 68.4%, 96.7%, and 100% of LSILs and 94.7%, 91.6%, and 100% of HSILs, respectively. Positive predictive values of these three biomarkers for HPV were 82.4%, 89.5%, and 91.4%, respectively. The positive predictive value for HPV of either cyclin E or p16 was 88.7%. Strong diffuse staining for p16 was significantly associated with high-risk HPV associated lesions. Normal or reactive epithelial changes scored positive for the three biomarkers in 7.7%, 8.0%, and 12%, respectively. Limitations in specificity included minimal or no suprabasal staining for Ki-67 in immature condylomas and occasional suprabasal staining of reactive epithelial changes (10%), diffuse weak nuclear cyclin E staining in some normal or metaplastic epithelia, and diffuse weak basal p16 staining and occasional stronger focal positivity in normal epithelia. Ki-67, cyclin E, and p16 are complementary surrogate biomarkers for HPV-related preinvasive squamous cervical disease. (Because cyclin E and p16 are most sensitive for LSIL and HSIL [including high-risk HPV], respectively, use of these biomarkers in combination for resolving diagnostic problems, with an appreciation of potential background staining, is recommended.) PMID- 11420460 TI - In situ adenocarcinoma of the bladder. AB - In situ adenocarcinoma of the bladder has not been well studied. Only one other case not associated with infiltrating adenocarcinoma has been reported in the literature. We identified 19 biopsies of in situ adenocarcinoma of the bladder without concurrent infiltrating adenocarcinoma or villous adenoma from the surgical pathology files of the Johns Hopkins Hospital between May 1984 and July 2000. The majority of patients (89%) were seen in consultation. The mean age at diagnosis was 70.4 years (range 48-88 years) and 79% were male. None of the patients developed a pure infiltrating adenocarcinoma; however, two patients had invasive urothelial carcinoma with focal glandular differentiation on prior or subsequent specimens. Two cases were pure in situ adenocarcinoma and 10 were seen with carcinoma in situ and/or papillary transitional cell cancer without invasion. Most patients (74%) had invasive carcinoma on either concurrent or subsequent specimens (five small cell and nine transitional cell [four micropapillary]). The majority (84%) of in situ adenocarcinomas were papillary, often seen in combination with either cribriform or flat architecture. In most cases the in situ adenocarcinoma was the predominant component when it was present with another in situ urothelial carcinoma. Seventy-nine percent of in situ adenocarcinomas showed >5 mitoses/10 HPF and 42% showed >10 mitoses/10 HPF. Moderate to severe nuclear pleomorphism was seen in 84% of cases. All cases showed apoptosis, and only one case showed focal necrosis. Seven patients were treated with cystectomy within 2-12 months. Of the other 12 patients, 10 were followed for a mean of 19.3 months (range 1-62 months). Ten (52%) patients were treated with bacille Calmette-Guerin, of whom four had no residual tumor on subsequent biopsy or cystectomy specimens. Three patients developed metastatic disease. In situ adenocarcinoma is a rare lesion that has a high incidence of association with small cell and micropapillary transitional cell carcinomas. When identified, in situ adenocarcinoma may indicate subsequent development of specific types of prognostically poor invasive carcinomas. PMID- 11420461 TI - Solitary fibrous tumor of the oral soft tissues: a clinicopathologic and immunohistochemical study of 16 cases. AB - We describe the clinicopathologic and immunohistochemical features of 16 solitary fibrous tumors (SFTs) originating within the oral soft tissues. There were 10 women and six men with a mean age at diagnosis of 56 years. In all cases the tumors were slow-growing, asymptomatic, submucosal growths of variable size and duration. The buccal mucosa was involved in 12 cases, and the tongue and lower lip were affected in two cases, respectively. Thirteen tumors developed on the left side of the mouth. Follow-up information was available in 10 cases and averaged 44.7 months, with no evidence of recurrence or metastasis. All 16 tumors were well circumscribed and demonstrated histologic features that were invariably benign. In all cases they were composed of an admixture of alternating hypercellular and hypocellular, collagenous zones; haphazardly arranged, cytologically bland spindle and ovoid cells that in areas were intimately intermixed with collagen; a prominent vasculature; and perivascular hyalinization. Immunohistochemically, the tumors were consistently positive for CD34, bcl-2, factor XIIIa, and vimentin, whereas 75% of the tumors were reactive for CD99. To further define the clinicopathologic profile of intraoral SFT, we include a review of the previously reported cases. We also include a comparison and brief review of the clinicopathologic and immunohistochemical features of other spindle cell tumors of the oral cavity, from which SFTs must be differentiated. PMID- 11420462 TI - Analysis of Hurthle cell neoplasms of the thyroid by interphase fluorescence in situ hybridization. AB - Recent studies have indicated that numerical chromosomal abnormalities including changes in p53 and cyclin D1 may be involved in Hurthle cell tumorigenesis. We analyzed a series of Hurthle cell neoplasms of the thyroid to evaluate the diagnostic and prognostic utility of numerical anomalies by DNA fluorescent probes for cyclin D1 and p53 gene loci and chromosomes 5, 7, 11, 12, 17, and 22. Interphase fluorescence in situ hybridization (FISH) analysis was performed on paraffin-embedded tissue sections from 10 Hurthle cell adenomas, 19 Hurthle cell carcinomas, and 7 normal thyroid tissues used as controls. Directly labeled fluorescent DNA probes for the centromere region of chromosomes 7, 11, 12, and 17 and locus-specific probes for chromosomes 5 and 22, cyclin D1, and p53 were utilized for dual-probe hybridizations. Sixty percent (6 of 10) Hurthle cell adenomas and 63% (12 of 19) Hurthle cell carcinomas showed chromosome gains. Twenty percent (2 of 10) Hurthle cell adenomas and 26% (5 of 19) Hurthle cell carcinomas showed chromosome losses. Normal thyroid tissues used as controls showed no chromosomal abnormalities. Among Hurthle cell tumors with chromosomal abnormalities, adenomas averaged 2.7 gains and 0.3 losses per case, and carcinomas averaged 3.3 gains and 0.6 losses per case. The two adenomas with chromosome losses each showed loss of one chromosome, whereas the five carcinomas with losses averaged 1.8 losses per case. Chromosome 22 was the most common loss identified, occurring in three of the 11 patients who died of disease. These results indicate that chromosomal imbalances as gains are common in both benign and malignant Hurthle cell neoplasms, but Hurthle cell carcinomas tend to have more chromosome losses than adenomas. Among Hurthle cell carcinomas in this study, chromosome losses were identified only from patients who died of disease. The loss of chromosome 22 may have prognostic value in Hurthle cell carcinoma of the thyroid. PMID- 11420463 TI - Adult neuroblastoma of the retroperitoneum and abdomen: clinicopathologic distinction from primitive neuroectodermal tumor. AB - Adult neuroblastoma (ANB) is a rare and poorly recognized entity among a histologically defined group of small, round-cell tumors arising in the retroperitoneum and abdomen. Eight cases of ANB were compared with seven cases of primitive neuroectodermal tumor (PNET) in these locations to identify clinicopathologic features that could be used to distinguish between the two lesions. The ANB study group included four men and four women 22-74 years of age (mean 38 years). Five patients with ANB presented with inflammatory symptoms or elevated levels of catecholamines and their metabolites. Five of the ANB tumors were classified as undifferentiated and three as poorly differentiated with a background of neuropil. These cases often showed immunoreactivity for multiple neural markers such as CD56, chromogranin A, synaptophysin, neurofilament, and neuron-specific enolase, but were negative for CD99, cytokeratins, desmin, myogenin, smooth muscle actin, muscle-specific actin, CD34, S-100 protein, and CD45. In contrast, all of the PNETs were positive for CD99, and four (57%) were also positive for cytokeratins. Two cases of ANB of the undifferentiated subtype had ultrastructural features characteristic of neuroblastoma and lacked a chimeric transcript (EWS-FLI1or ERG), which is specific for PNET. All five patients with the undifferentiated subtype of ANB and six of the seven patients with PNET died of their disease within 3 years of discovery of the lesion. Our results show that ANB, although rare, should be considered in the differential diagnosis of patients with small, round-cell tumors in the retroperitoneum and abdomen. Appropriate immunohistochemical studies and laboratory examination enable pathologists to distinguish ANB from other differential diagnoses, especially PNET. PMID- 11420464 TI - TARC, a CC chemokine, is frequently expressed in classic Hodgkin's lymphoma but not in NLP Hodgkin's lymphoma, T-cell-rich B-cell lymphoma, and most cases of anaplastic large cell lymphoma. AB - Thymus and activation-regulated chemokine (TARC) has been identified as a lymphocyte-directed CC chemokine that attracts activated T-helper type 2 (Th2) cells in humans. Recent studies showed that the T cells surrounding Reed Sternberg cells in Hodgkin's lymphomas (HL) are Th2 type. Anaplastic large cell lymphomas (ALCL), T-cell-rich B-cell lymphoma (TCRBCL) can mimic HL in some instances. This study aimed to establish the pattern of TARC expression in these diseases. Immunohistochemical stain using a polyclonal goat anti-human antibody to TARC was performed on 119 cases of confirmed HL; 99 were classical type (43 mixed cellularity, 43 nodular sclerosis, 5 lymphocyte depleted, 4 lymphocyte rich, 4 unclassifiable) and 20 lymphocyte predominant HL. Additional 27 ALCL (9 T , 18 null-cell phenotype), 16 T-cell and 8 B-cell non-Hodgkin's lymphoma (NHL) were studied. A total of 85.8% of the classical HL, one case of ALCL, and one case of large cell B-cell lymphoma with anaplastic morphology showed positive TARC expression in the tumor cells. The expression was paranuclear and/or diffuse in the cell cytoplasm. The tumor cells in all cases of lymphocyte predominant HL, TCRBCL, null ALCL, and T-NHL did not express TARC. The high frequency of TARC expression in the Reed-Sternberg cells of classical HL may explain the characteristic T-cell infiltrate in this disease. The absence in other types that may be morphologically similar indicates that staining for TARC may aid in differential diagnosis. PMID- 11420465 TI - Prospective multicenter evaluation of the morphometric D-score for prediction of the outcome of endometrial hyperplasias. AB - Prospective multicenter evaluation of the WHO classification and the morphometric D-score to predict endometrial hyperplasia cancer progression. In 132 endometrial hyperplasias WHO classification was performed by two experienced gynecologic pathologists. The D-score was assessed blindly by technicians in a routine diagnostic setting. Development of endometrial carcinoma during a 1-10-year follow-up was used as the end point. Eleven of 132 patients (8%), 10 of 61 (16%) atypical hyperplasias, and 1 of 71 (1%) nonatypical hyperplasias developed cancer. Twenty-six curettings had a D-score < or = 0 ("unfavorable" or endometrial intraepithelial neoplasia) of which 10 (38%) developed cancer. None of the 86 cases with a D-score > 1 ("favorable") and one of the 20 (5%) cases with 0 < D-score < or = 1 ("uncertain") developed cancer. Sensitivity of the D score was 100%, specificity 82%, the positive and negative predictive values were 38% and 100%, respectively. These values are similar to those in three prior retrospective D-score studies but higher than the WHO values (which are 91%, 58%, 16%, and 99%, respectively). The D-score in endometrial hyperplasias is a more sensitive and specific marker for cancer prediction than the WHO classification, can be assessed in a routine clinical setting on standard hematoxylin and eosin sections (15-30 minutes per case), and is highly reproducible and cost-effective (U.S. $50 per case). PMID- 11420466 TI - Molecular pathologic analysis enhances the diagnosis and management of Muir-Torre syndrome and gives insight into its underlying molecular pathogenesis. AB - The Muir-Torre syndrome (MTS) is an autosomal dominantly inherited disorder, characterized by visceral malignancies and sebaceous skin lesions. In a subset of MTS families the disease is due to an underlying DNA mismatch-repair defect. We have identified a MTS family whose spectrum of reported neoplasia included adenocarcinomas of numerous gastrointestinal sites, carcinomas of the endometrium, ovary and breast, papillary transitional cell carcinoma of the ureter, a range of cutaneous tumors, as well as keratoacanthomas. All tumors were tested for microsatellite instability and immunohistochemically stained for expression of MLH1 and MSH2 proteins. All tumors were found to be microsatellite unstable and lacking in MSH2 protein expression. The subsequent mutation detection focused on hMSH2, and a germline mutation was identified (CAA-->TAA, Gln-->STOP, codon 337). This mutation was subsequently found in a family member with a single skin lesion only. We propose that the combination of immunohistologic and microsatellite instability analysis can be exploited to screen individuals with characteristic skin lesions even before development of visceral tumors and to direct the subsequent germline mutation search. The profile of microsatellite instability and the genes rendered dysfunctional differed between tumor samples, suggesting that the molecular pathogenesis varied between lesions, despite a common germline mutation. PMID- 11420467 TI - The immunohistochemical mucin expression pattern distinguishes different types of intraductal papillary mucinous neoplasms of the pancreas and determines their relationship to mucinous noncystic carcinoma and ductal adenocarcinoma. AB - Intraductal papillary-mucinous neoplasms of the pancreas seem to comprise various types, whose relationship to ductal adenocarcinoma and mucinous noncystic carcinoma is unclear. We analyzed the mucin immunophenotype and the DPC4/SMAD4 expression in intraductal papillary-mucinous neoplasms, ductal carcinomas, and mucinous noncystic carcinomas to define features that may help to distinguish between different types of intraductal papillary-mucinous neoplasms and to establish their relationship to other neoplasms of the exocrine pancreas. A series of 51 intraductal papillary-mucinous neoplasms, three mucinous noncystic carcinomas (two with an intraductal component), and 35 ductal adenocarcinomas were screened immunohistochemically for their expression of MUC1, MUC2, MUC5, and DPC4/SMAD4. All intraductal papillary-mucinous neoplasms and mucinous noncystic carcinomas were positive for MUC5. Thirty-two intraductal papillary-mucinous neoplasms and three mucinous noncystic carcinomas abundantly expressed MUC2 but no (or only little) MUC1. The remaining intraductal papillary-mucinous neoplasms showed either mainly MUC1 expression or focal MUC1 and MUC2 expression. All ductal carcinomas but one were MUC2 negative and MUC1 and MUC5 positive. DPC4 was not expressed in two intraductal papillary-mucinous neoplasms that showed a tubular invasion pattern. Twelve of 23 ductal adenocarcinomas were DPC4 positive. Intraductal papillary-mucinous neoplasms can be divided into at least three different mucin immunophenotypes. The first and largest group of intraductal papillary-mucinous neoplasms and mucinous noncystic carcinomas is MUC1 negative and MUC2 positive and probably forms one tumor entity. The second group seems to be related to ductal carcinoma because of its MUC1 positivity in the absence or very weak MUC2 staining. The third group shows focal MUC1/MUC2 expression and is characterized by oncocytic histology. PMID- 11420468 TI - SHP-1 expression by malignant small B-cell lymphomas reflects the maturation stage of their normal B-cell counterparts. AB - SHP-1 is a protein phosphotyrosine phosphatase that plays an important role in modulating intracellular signaling, which regulates cell activation, proliferation, differentiation, and migration. It is a negative regulator of signal transduction induced by a number of cell receptors. Our immunohistochemical examination of paraffin-embedded reactive lymph nodes and lymphoid tissues revealed that B lymphocytes in follicle germinal centers do not express SHP-1. A weak staining of the B cells in the germinal center light zones was detected when an ultrasensitive amplification system was used. In contrast, normal B cells in mantle and marginal zones as well as interfollicular B lymphocytes and plasma cells displayed strong immunoreactivity. This pattern of SHP-1 expression was repeated in small B-cell lymphomas. All cases of mantle cell lymphoma (12 of 12), marginal zone lymphoma (10 of 10), and chronic lymphocytic leukemia/small lymphocytic lymphoma (13 of 13) expressed SHP-1 protein. However, only 1 of 30 cases of grade 1 follicle center cell lymphoma expressed SHP-1. Our observations highlight the biologic functions of SHP-1 and demonstrate that the SHP-1 expression pattern by small B-cell lymphomas reflects the maturation stage of their normal cell counterparts. These results indicate that determination of SHP-1 expression may help in the differential diagnosis of small B-cell lymphomas. PMID- 11420469 TI - Atypical bile duct adenoma, clear cell type: a previously undescribed tumor of the liver. AB - A variable proportion of bile duct adenomas of the liver are still confused with metastatic well-differentiated adenocarcinoma by surgeons and pathologists. We present here three examples of previously undescribed primary hepatic bile duct tumors that were composed almost entirely of clear cells that closely mimicked metastatic renal cell carcinoma. They were interpreted as atypical bile duct adenomas and occurred in two males and one female whose ages ranged from 25 to 64 years. All three tumors were incidental findings and measured from 0.8 to 1.1 cm. The clear neoplastic cells showed mild nuclear atypia and no mitotic activity. They were arranged in tubules and nests that focally infiltrated the hepatic parenchyma. For comparison, a case of clear cell cholangiocarcinoma and 13 conventional bile duct adenomas were examined. The clear cell cholangiocarcinoma was larger (6.0 cm) and had the tubular pattern of conventional cholangiocarcinoma and an abundant desmoplastic stroma. The clear cells of this tumor exhibited greater nuclear atypia and increased mitotic activity. All three atypical bile duct adenomas expressed cytokeratin (CK) 7, p53 protein, epithelial membrane antigen (EMA), and carcinoembryonic antigen (CEA); they were negative for CK20, vimentin, Hep Par 1, chromogranin, and prostatic specific antigen (PSA) and exhibited less than 10% of Ki-67-positive nuclei. One atypical bile duct adenoma displayed luminal immunoreactivity for villin. With the exception of Ki 67 reactivity, the 13 conventional bile duct adenomas and the clear cell cholangiocarcinoma had essentially a similar immunohistochemical profile as that of the atypical clear cell bile duct adenomas. The absence of an extrahepatic primary tumor, the histologic features, the immunohistochemical profile, and the fact that all patients are symptom-free 2 months to 18 years after wedge liver biopsy support the interpretation of atypical clear cell bile duct adenoma. The differential diagnosis with clear cell hepatocellular carcinoma and metastatic clear cell carcinomas is discussed. PMID- 11420470 TI - ADASP recommendations for processing and reporting lymph node specimens submitted for evaluation of metastatic disease. PMID- 11420471 TI - Rhabdoid papillary meningioma. AB - We have studied an uncommon case of rhabdoid papillary meningioma in a 15-year old boy with a dura-based mass arising in the left posterior fossa. The patient exhibited prominent extracranial extension during the past 6 years, consisting of a mixture of both perivascular pseudopapillary growth and rhabdoid cytologic features of neoplastic meningothelial cells. The meningothelial features were evidenced by the focal whorl formation of tumor cells, coexpression of epithelial membrane antigen and vimentin, and ultrastructural findings of interdigitated cytoplasmic process and intercellular junction. However, the regional and histologic resemblances to ependymoma were further complicated by unexpected focal expression of glial fibrillary acidic protein, neurofilament, and alpha smooth muscle actin of the tumor cells. The rhabdoid morphology was characterized by sheets of tumor cells with eccentric nuclei and abundant eosinophilic cytoplasm with often recognizable intracytoplasmic hyaline inclusions. These inclusions revealed ultrastructural paranuclear whorls of intermediate filaments, ruling out the other forms of intracytoplasmic eosinophilic inclusions resembling rhabdoid morphology. Diagnosis of an unusual rhabdoid papillary meningioma with aggressive behavior is resoluble by immunohistochemical and ultrastructural analyses. PMID- 11420472 TI - Nuchal-type fibroma is positive for CD34 and CD99. PMID- 11420473 TI - Hepatosplenic T-cell lymphoma of alphabeta lineage. PMID- 11420474 TI - Synovial sarcoma in the kidney. PMID- 11420475 TI - Renal angiomyolipoma. A clinicopathologic, immunohistochemical, and follow-up study of 46 cases. PMID- 11420476 TI - Telerobotic laparoscopic cholecystectomy: initial clinical experience with 25 patients. AB - OBJECTIVE: To determine the safety and feasibility of performing telerobotic laparoscopic cholecystectomies. This will serve as a preliminary step toward the integration of computer-rendered three-dimensional preoperative imaging studies of anatomy and pathology onto the patient's own anatomy during surgery. SUMMARY BACKGROUND DATA: Computer-assisted surgery (CAS) increases the surgeon's dexterity and precision during minimally invasive surgery, especially when using microinstruments. Clinical trials have shown the improved microsurgical precision afforded by CAS in the minimally invasive setting in cardiac and gynecologic surgery. Future applications would allow integration of preoperative data and augmented-reality simulation onto the actual procedure. METHODS: Beginning in September 1999, CAS was used to perform cholecystectomies on 25 patients at a single medical center in this nonrandomized, prospective study. The operations were performed by one of two surgeons who had previous laboratory experience using the computer interface. The entire dissection was performed by the surgeon, who remained at a distance from the patient but in the same operating room. The operation was evaluated according to time of dissection, time of assembly/disassembly of robot, complications, immediate postoperative course, and short-term follow-up. RESULTS: Twenty of the 25 patients had symptomatic cholelithiasis, 1 had a gallbladder polyp, and 4 had acute cholecystitis. Twenty four of the 25 laparoscopic cholecystectomies were successfully completed by CAS. There was one conversion to conventional laparoscopic cholecystectomy. Set-up and takedown of the robotic arms took a median of 18 minutes. The median operative time for dissection and the overall operative time were 25 and 108 minutes, respectively. There were no intraoperative complications. There was one postoperative complication of a suspected pulmonary embolus, which was treated with anticoagulation. All patients were tolerating diet at discharge. CONCLUSIONS: Laparoscopic cholecystectomy performed by CAS is safe and feasible, with operative times and patient recovery similar to those of conventional laparoscopy. At present, CAS cholecystectomy offers no obvious advantages to patients, but the potential advantages of CAS lie in its ability to convert the surgical act into digitized data. This digitized format can then interface with other forms of digitized data, such as pre- or intraoperative imaging studies, or be transmitted over a distance. This has the potential to revolutionize the way surgery is performed. PMID- 11420477 TI - Surgery of the 21st century. PMID- 11420478 TI - DNA methylation: an alternative pathway to cancer. AB - OBJECTIVE: To provide an introduction to the concept of DNA methylation and its function in normal cells, and to explain the possible mechanisms as to how abnormalities in this phenomenon can relate to carcinogenesis. The clinical implications with reference to common malignancies encountered in surgical practice are discussed. SUMMARY BACKGROUND DATA: Methylation of DNA is a heritable, enzyme-induced modification to DNA structure without alteration of the specific sequence of the base pairs responsible for encoding the genome. DNA methylation can both directly inhibit the expression of genes and also increase the probability that affected genes undergo a mutational event. Although DNA methylation plays an essential role in normal biologic processes, distinct and abnormal patterns of methylation are observed in cancers. In particular, there has been increased documentation that methylation of the promoter regions of several genes, including known tumor suppressor genes, results in the subsequent failure to express their functional proteins. Consequently, DNA methylation may represent an early and fundamental step in the pathway by which normal tissue undergoes neoplastic transformation. Further, an assessment of the methylation profiles within neoplastic tissues may provide key information in enhancing the diagnosis, predicting the clinical behavior, and designing specific treatment plans for individual patients. METHODS: Published literature from 1925 to 2000 contributing to an understanding of the purpose of DNA methylation and how pathology of this phenomenon could contribute to cancer are reviewed. Theories on these issues and the evidence that led to them are described. The present status of the subject in a clinical context is discussed. RESULTS: Gene expression can be significantly modulated by alterations in DNA methylation patterns. Methylation within the promoter regions of tumor suppressor genes causes their silencing, and methylation within the gene itself can induce mutational events. These mechanisms may play a fundamental role in precipitating the development of a large and diverse number of human cancers. CONCLUSIONS: DNA methylation is an important factor in the development of cancer. A greater understanding of the relationship between DNA methylation events at the molecular level and its interaction in the clinical context may provide the basis for future advances in the surgical and pharmacologic management of malignant diseases. PMID- 11420479 TI - Sutureless closed hemorrhoidectomy: a new technique. AB - OBJECTIVE: To compare a new technique of radical hemorrhoidectomy using an electrothermal device originally devised to seal vessels in abdominal operations, with the conventional open Milligan-Morgan procedure performed with diathermy. SUMMARY BACKGROUND DATA: Hemorrhoidectomy is one of the most commonly performed anorectal operations. Two well-established methods, the "open" Milligan-Morgan excision and the "closed" Ferguson technique, both carry risks of postoperative bleeding, urinary retention, and late anal stenosis. The convalescence is similarly long and difficult after both operations. The quest for an improved technique of radical excision of hemorrhoids is justified. METHODS: In this case control study, two groups of patients were alternatively allocated into study and control groups. In the study group (n = 40), an electrothermal system was used. The tissue fusion produced by this device consists of melting of collagen and elastin. This technique essentially achieves a sutureless closed hemorrhoidectomy. The operative time, postoperative complications, and time off work were compared with the group undergoing conventional Milligan-Morgan hemorrhoidectomy (control group, n = 40). RESULTS: The operative time and time off work were significantly shorter in the study group. There were also fewer postoperative complications in this group. CONCLUSIONS: The "tissue-welding" properties of this device and the shape of the electrode handpiece may be successfully applied to the performance of an operation most appropriately described as a "modified sutureless closed hemorrhoidectomy." This pilot study shows that this new technique is simple and safe, significantly shortens the operation, and is followed by a significantly easier and shorter recovery. PMID- 11420480 TI - Skeletonizing en bloc esophagectomy for cancer. AB - OBJECTIVE: To evaluate the long-term outcome of patients with esophageal cancer after resection of the extraesophageal component of the neoplastic process en bloc with the esophageal tube. SUMMARY BACKGROUND DATA: Opinions are conflicting about the addition of extended resection of locoregional lymph nodes and soft tissue to removal of the esophageal tube. METHODS: Esophagectomy performed en bloc with locoregional lymph nodes and resulting in a real skeletonization of the nonresectable anatomical structures adjacent to the esophagus was attempted in 324 patients. The esophagus was removed using a right thoracic (n = 208), transdiaphragmatic (n = 39), or left thoracic (n = 77) approach. Lymphadenectomy was performed in the upper abdomen and lower mediastinum in all patients. It was extended over the upper mediastinum when a right thoracic approach was used and up to the neck in 17 patients. Esophagectomy was carried out flush with the esophageal wall as soon as it became obvious that a macroscopically complete resection was not feasible. Neoplastic processes were classified according to completeness of the resection, depth of wall penetration, and lymph node involvement. RESULTS: Skeletonizing en bloc esophagectomy was feasible in 235 of the 324 patients (73%). The 5-year survival rate, including in-hospital deaths (5%), was 35% (324 patients); it was 64% in the 117 patients with an intramural neoplastic process versus 19% in the 207 patients having neoplastic tissue outside the esophageal wall or surgical margins (P <.0001). The latter 19% represented 12% of the whole series. The 5-year survival rate after skeletonizing en bloc esophagectomy was 49% (235 patients), 49% for squamous cell versus 47% for glandular carcinomas (P =.4599), 64% for patients with an intramural tumor versus 34% for those with extraesophageal neoplastic tissue (P <.0001), and 43% for patients with fewer than five metastatic nodes versus 11% for those with involvement of five or more lymph nodes (P =.0001). CONCLUSIONS: The strategy of attempting skeletonizing en bloc esophagectomy in all patients offers long-term survival to one third of the patients with resectable extraesophageal neoplastic tissues. These patients represent 12% of the patients with esophageal cancer suitable for esophagectomy and 19% of those having neoplastic tissue outside the esophageal wall or surgical margins. PMID- 11420481 TI - Patient evaluation and management with selective use of magnetic resonance cholangiography and endoscopic retrograde cholangiopancreatography before laparoscopic cholecystectomy. AB - OBJECTIVE: To assess the utility of triage guidelines for patients with cholelithiasis and suspected choledocholithiasis, incorporating selective use of magnetic resonance cholangiography (MRC) and endoscopic retrograde cholangiopancreatography (ERCP) before laparoscopic cholecystectomy (LC). SUMMARY BACKGROUND DATA: ERCP is the most frequently used modality for the diagnosis and resolution of choledocholithiasis before LC. MRC has recently emerged as an accurate, noninvasive modality for the detection of choledocholithiasis. However, useful strategies for implementing this diagnostic modality for patient evaluation before LC have not been investigated. METHODS: During a 16-month period, the authors prospectively evaluated all patients before LC using triage guidelines incorporating patient information obtained from clinical evaluation, serum chemistry analysis, and abdominal ultrasonography. Patients were then assigned to one of four groups based on the level of suspicion for choledocholithiasis (group I, extremely high; group 2, high; group 3, moderate; group 4, low). Group 1 patients underwent ERCP and clearance of common bile duct stones; group 2 patients underwent MRC; group 3 patients underwent LC with intraoperative cholangiography; and group 4 patients underwent LC without intraoperative cholangiography. RESULTS: Choledocholithiasis was detected in 43 of 440 patients (9.8%). The occurrence of choledocholithiasis among patients in the four groups were 92.6% (25/27), 32.4% (12/37), 3.8% (2/52), and 0.9% (3/324) for groups 1, 2, 3, and 4, respectively (P <.001). MRC was used for 8.4% (37/440) of patients. Patient triage resulted in the identification of common bile duct stones during preoperative ERCP in 92.3% (36/39) of the patients. Unsuspected common bile duct stones occurred in six patients (1.4%). CONCLUSIONS: The probability of choledocholithiasis can be accurately assessed based on information obtained during the initial noninvasive evaluation. Stratification of risks for choledocholithiasis facilitates patient management with the most appropriate diagnostic studies and interventions, thereby improving patient care and resource utilization. PMID- 11420482 TI - Effect of preoperative biliary decompression on pancreaticoduodenectomy associated morbidity in 300 consecutive patients. AB - OBJECTIVE: To examine the relationship between preoperative biliary drainage and the morbidity and mortality associated with pancreaticoduodenectomy. SUMMARY BACKGROUND DATA: Recent reports have suggested that preoperative biliary drainage increases the perioperative morbidity and mortality rates of pancreaticoduodenectomy. METHODS: Peri-operative morbidity and mortality were evaluated in 300 consecutive patients who underwent pancreaticoduodenectomy. Univariate and multivariate logistic regression analyses were done to evaluate the relationship between preoperative biliary decompression and the following end points: any complication, any major complication, infectious complications, intraabdominal abscess, pancreaticojejunal anastomotic leak, wound infection, and postoperative death. RESULTS: Preoperative prosthetic biliary drainage was performed in 172 patients (57%) (stent group), 35 patients (12%) underwent surgical biliary bypass performed during prereferral laparotomy, and the remaining 93 patients (31%) (no-stent group) did not undergo any form of preoperative biliary decompression. The overall surgical death rate was 1% (four patients); the number of deaths was too small for multivariate analysis. By multivariate logistic regression, no differences were found between the stent and no-stent groups in the incidence of all complications, major complications, infectious complications, intraabdominal abscess, or pancreaticojejunal anastomotic leak. Wound infections were more common in the stent group than the no-stent group. CONCLUSIONS: Preoperative biliary decompression increases the risk for postoperative wound infections after pancreaticoduodenectomy. However, there was no increase in the risk of major postoperative complications or death associated with preoperative stent placement. Patients with extrahepatic biliary obstruction do not necessarily require immediate laparotomy to undergo pancreaticoduodenectomy with acceptable morbidity and mortality rates; such patients can be treated by endoscopic biliary drainage without concern for increased major complications and death associated with subsequent pancreaticoduodenectomy. PMID- 11420483 TI - Value of lipiodol computed tomography and digital subtraction angiography in the era of helical biphasic computed tomography as preoperative assessment of hepatocellular carcinoma. AB - OBJECTIVE: To compare the diagnostic accuracies of Lipiodol computed tomography (CT) and helical biphasic CT as preoperative imaging modalities for hepatocellular carcinoma (HCC). SUMMARY BACKGROUND DATA: Lipiodol CT after digital subtraction angiography has long been used as a highly sensitive imaging modality for HCC. The recent advent of helical CT has allowed scanning the entire liver during both the arterial and portal venous phase of contrast enhancement. METHODS: The authors analyzed data from 164 patients who underwent hepatic resection for HCC to calculate the sensitivity and specificity of these modalities. Findings of intraoperative ultrasonography followed by histologic confirmation were set as the gold standard. RESULTS: Although sensitivity decreased with both modalities as tumors became small and well differentiated, helical CT showed a higher sensitivity than Lipiodol CT in detecting well differentiated HCC nodules smaller than 2 cm. In contrast, Lipiodol CT was superior to helical CT for the detection of small but moderately to poorly differentiated nodules. The overall sensitivity of helical CT was higher than that of Lipiodol CT. These findings suggest that helical CT is superior in delineating early HCC, whereas Lipiodol CT is specific to the detection of intrahepatic metastases. In terms of specificity, helical CT was superior to Lipiodol CT. CONCLUSIONS: Helical CT and Lipiodol CT are complementary modalities. At present, helical biphasic CT does not obviate the need for invasive techniques such as angiography and Lipiodol CT as preoperative examinations for HCC. PMID- 11420484 TI - Improving survival results after resection of hepatocellular carcinoma: a prospective study of 377 patients over 10 years. AB - OBJECTIVE: To investigate whether the survival results after resection of hepatocellular carcinoma (HCC) have improved within the past decade by an analysis of a prospective cohort of patients over a 10-year period. SUMMARY BACKGROUND DATA: The surgical death rate after resection of HCC has greatly improved in recent years, but the long-term prognosis remains unsatisfactory. It remains unknown whether the survival results after resection of HCC have improved within the past decade. METHODS: The clinicopathologic and follow-up data of 377 patients who underwent curative resection of HCC between January 1989 and January 1999 were prospectively collected. These patients were categorized according to two time periods: before 1994 (group 1, n = 136) and after 1994 (group 2, n = 241). The two groups were compared for clinicopathologic data and survival results. The prognostic factors for disease-free survival were further analyzed to identify the factors that might have led to improved survival outcomes. RESULTS: The overall and disease-free survival results were significantly better in group 2 compared with group 1. Patients in group 2 had significantly higher proportions of subclinical presentation, small tumors, and tumors of early pTNM stage. There were also significantly lower frequencies of histologic margin involvement, less intraoperative blood loss, and a lower transfusion rate in group 2. By multivariate analysis, early pTNM stage, subclinical HCC, and no perioperative transfusion were independent favorable prognostic factors for disease-free survival. CONCLUSIONS: Significant improvement of overall and disease-free survival results after resection of HCC has been achieved within the past decade as a result of advances in the diagnosis and surgical management of HCC. Earlier diagnosis of HCC by better imaging modalities, increased detection of subclinical HCC by screening of high-risk patients, and a reduced perioperative transfusion rate were identified as the major contributory factors for the improved outcomes. PMID- 11420485 TI - Improved results of liver resection for hepatocellular carcinoma on cirrhosis give the procedure added value. AB - OBJECTIVE: To review a single-center experience to update the performance indexes of liver resection (LR). SUMMARY BACKGROUND DATA: Several therapies have been proposed in the treatment of hepatocellular carcinoma (HCC) on cirrhosis, although LR was the first to be widely applied. METHODS: Of 408 patients with cirrhosis admitted for HCC in the period 1983 to 1998, 264 had a LR. Patient selection, surgical technique, 30-day deaths, long-term survival, recurrence rate, and recurrence treatment were reviewed after stratifying patients according to the year of surgery. Mean follow-up was 34.5 +/- 29.1 months. RESULTS: The number of Child A patients who underwent surgery after the discovery of the tumor at routine evaluation increased significantly from 64.5% to 87.9% during the study period. Procedures carried out without blood transfusions increased from 31.4% to 76.9%. The overall operative death rate was 4.9%. Actuarial survival rates were 63.1% and 41.1% after 3 and 5 years, respectively; actuarial tumor free survival rates were 49.3% and 27.9% at the same intervals. After 1992, surgical deaths decreased from 9.3% to 1.3%. Actuarial survival rates increased from 52.9% and 32.3% to 71.7% and 49.4% after 3 and 5 years, respectively. There was no difference in the actuarial recurrence rate between the two periods, but the chance to treat recurrence increased over time from 22.4% to 53.7% with a concomitant, significant improvement in survival. CONCLUSIONS: LR represents a well-established therapy for HCC on cirrhosis. It remains one of the fundamentals in the multidisciplinary approach to this tumor and should be considered as the first option for patients with preserved hepatic function and limited disease. Today, LR should offer a surgical death rate of less than 1.5%, a 5-year survival rate of approximately 50%, and a 5-year tumor-free survival rate of 28% when performed in specialized centers. PMID- 11420486 TI - Halsted revisited: internal mammary sentinel lymph node biopsy in breast cancer. AB - OBJECTIVE: To investigate the feasibility of internal mammary sentinel lymph node biopsy as a method to refine and thereby improve nodal staging in breast cancer. SUMMARY BACKGROUND DATA: The internal mammary lymph node status is a major prognostic factor in breast cancer. If positive, prognosis is less favorable. However, staging this regional nodal basin is not performed routinely, thus discarding additional staging information. METHODS: In a consecutive series of 256 patients with primary breast cancer, sentinel node biopsy was performed based on lymphoscintigraphy, intraoperative gamma probe detection, and blue dye mapping using 10 mCi (370 MBq) (99m)Tc-nanocolloid injected peritumorally and 0.5 to 1.0 mL Patent Blue V injected intradermally. During surgery, whenever possible, both axillary and internal mammary sentinel nodes were sampled. RESULTS: Lymphoscintigraphy showed axillary sentinel nodes in 95% (243/256) and additional internal mammary sentinel nodes in 25.3% (65/256). The overall success rate of axillary sentinel node biopsy was 97% (249/256). Sampling the internal mammary basin, based on the results of lymphoscintigraphy, was successful in 63% (41/65). In three patients a small pleural lesion resulted from staging this basin. This technique revealed internal mammary metastases in 26.8% (11/41). In 7.3% (3/41), internal mammary nodes showed metastatic involvement without accompanying axillary metastases. CONCLUSIONS: Internal mammary sentinel node biopsy is feasible without serious additional complications. It improves nodal staging in breast cancer by identifying higher-risk subgroups with internal mammary nodal metastases, which might benefit from altered adjuvant treatment regimens. PMID- 11420487 TI - Expression of adrenocorticotropin receptor gene in adrenocortical adenomas from patients with Cushing syndrome: possible contribution for the autonomous production of cortisol. AB - OBJECTIVE: To examine whether inhibition of endogenous adrenocorticotropin (ACTH) secretion in patients with Cushing syndrome affects the expression of the ACTH receptor (ACTH-R) gene in adrenocortical adenoma and attached atrophic normal gland. SUMMARY BACKGROUND DATA: ACTH increases adrenal cell growth and steroidogenesis by means of ACTH-R. In vivo and in vitro studies have shown that expression of ACTH-R is upregulated by its own ligand ACTH in several species. In patients with Cushing syndrome resulting from adrenocortical adenoma, there is autonomous production of cortisol from the adenoma. This strongly inhibits endogenous ACTH secretion, giving rise to the speculation that the expression of the ACTH-R gene in these patients is also suppressed. However, previous studies have shown that administration of exogenous ACTH to these patients leads to a further increase in the production of cortisol, suggesting the expression of functional ACTH-R in the adenoma. The authors, therefore, examined the expression of the ACTH-R gene in these patients. METHODS: Fourteen patients with Cushing syndrome were studied. Glucocorticoid excess resulting from autonomous production from the adenomas was ascertained, and unilateral adrenalectomy was performed. The levels of ACTH-R and cytochrome P450 side chain cleavage enzyme (P450scc) mRNAs were determined by Northern blot analysis. The entire coding region of the ACTH-R gene in these patients was sequenced. RESULTS: ACTH-R mRNA abundance in the attached atrophic normal adrenals was suppressed and invariably less than that in the normal gland obtained from a patient with renal cancer. However, the expression of ACTH-R mRNA was not suppressed in any of the adenomas. Expression of ACTH-R mRNA in the adenomas was four- to sixfold greater than that in the attached atrophic gland. No mutation in the coding sequence of the ACTH-R gene in the adenoma was detected in any of the patients. The mRNA in the adenomas appeared to be translated into functionally active receptor because intramuscular administration of ACTH resulted in significant increases in plasma cortisol before surgery but not 3 months after surgery. In addition, there was a positive linear correlation between the expressions of ACTH-R and P450scc mRNAs in the adenoma tissue. CONCLUSIONS: Suppressed ACTH secretion in patients with Cushing syndrome results in reduction of the ACTH-R mRNA expression in nonneoplastic adrenocortical cells. However, the regulatory mechanism of ACTH-R expression might be different in adenoma. Persistent expression in the adenoma of ACTH-R alone, even in the absence of ACTH, might result in increased basal adenyl cyclase activity, as observed in the case of thyroid-stimulating hormone receptor, and thereby might play a role in the autonomous production of cortisol. PMID- 11420488 TI - Oral feeding with glutamine prevents lymphocyte and glutathione depletion of Peyer's patches in endotoxemic mice. AB - OBJECTIVE: To determine the effect of oral glutamine feeding on lymphocyte subpopulations and glutathione metabolism in Peyer's patches (PPs) of healthy and endotoxemic mice. SUMMARY BACKGROUND DATA: Recent data indicate that nutrients both maintain nitrogen and energy balances and modulate cell and organ function. In particular, glutamine has an impact on gut and immune function. This is of special importance in the perioperative phase. METHODS: Female Balb/c mice were fed a glutamine-enriched diet or a control diet for 10 days. On day 7 25 microg lipopolysaccharide (LPS) or saline was injected. On day 3 after the challenge, mice were killed, total cell yield was determined, and lymphocyte subpopulations (total T cells, CD4+, CD8+ cells, and B cells) were analyzed by flow cytometry. One experimental group was treated with buthionine sulfoximine, a specific inhibitor of glutathione synthesis. The glutathione content in PPs was measured by high-performance liquid chromatography. RESULTS: Glutamine administration led to a significant increase in total cell yield, including T and B cells, in PPs. The LPS-induced reduction of T cells (-45%) and of B cells (-30%) was significantly lower in glutamine-treated mice. Endotoxemia caused a 42% decrease of glutathione in control animals, but not in glutamine-treated animals. As with LPS, buthionine sulfoximine also lowered lymphocyte numbers and glutathione content of the PPs. CONCLUSIONS: Administration of glutamine prevents LPS stimulated lymphocyte atrophy in PPs, possibly by increasing the glutathione content in the PPs. Therefore, oral glutamine supply seems to be a suitable approach for improving intestinal immunity in immunocompromised patients. PMID- 11420489 TI - Protective effect of ischemic preconditioning on cold preservation and reperfusion injury associated with rat intestinal transplantation. AB - OBJECTIVE: To define the protective effect of ischemic preconditioning on cold ischemia and reperfusion injury associated with intestinal transplantation, and the role of nitric oxide in this process. SUMMARY BACKGROUND DATA: Ischemia/reperfusion injury continues to be a significant obstacle in small bowel transplantation. Preconditioning is a mechanism that protects against this injury. METHODS: To study the capacity of preconditioning to prevent cold ischemia-associated injury and the inflammatory response associated with intestinal transplantation, the authors studied a control group of animals, cold ischemia groups with or without previous preconditioning and with or without previous administration of L-NAME or NONOS, and intestinal transplantation groups with or without previous preconditioning and with or without previous administration of L-NAME or NONOS. RESULTS: Histologic findings and the release of lactate dehydrogenase into the preservation solution showed that preconditioning protects against cold ischemic preservation-associated injury. Preconditioning also prevented the inflammatory response associated with intestinal transplantation, measured by the above parameters and by neutrophil recruitment in the intestine. Inhibition of nitric oxide eliminates the protective effect. CONCLUSIONS: Preconditioning protects the intestinal grafts from cold preservation and reperfusion injury in the rat intestinal transplantation model. Nitric oxide is involved in this protection. PMID- 11420490 TI - Effect of prolonged selective intramesenteric arterial vasodilator therapy on intestinal viability after acute segmental mesenteric vascular occlusion. AB - OBJECTIVE: To evaluate the effect of selective intramesenteric artery vasodilator infusion on intestinal viability in a rat model of acute segmental mesenteric vascular occlusion. SUMMARY BACKGROUND DATA: Although intramesenteric arterial vasodilator infusion may be an effective treatment for nonocclusive mesenteric ischemia, it has also been advocated to increase collateral blood flow after mesenteric vascular occlusion. However, the authors have previously found that intraarterial vasodilators actually reduce collateral blood flow acutely, by preferentially dilating the vasculature of adjacent, nonischemic mesenteric vascular beds, a phenomenon well established in other organs. METHODS: A segment of rat ileum was acutely devascularized, with blood flow provided only by collateral arterial vessels from adjacent, nonischemic bowel. Papaverine (30 or 40 microg/kg/min), isoproterenol (0.06 microg/kg/min), norepinephrine (0.1 or 0.2 microg/kg/min), or vehicle saline was continuously infused into the cranial (superior) mesenteric artery for 48 hours. Viability was then assessed using previously established, objective gross and microscopic criteria. RESULTS: Although papaverine increased total mesenteric blood flow in normally vascularized rats, it not only failed to improve but actually significantly reduced the length of the devascularized segment maintained viable by collateral blood flow after 48 hours. Isoproterenol had a similar effect. Norepinephrine infusion decreased both normal mesenteric blood flow and viable segment length. CONCLUSIONS: These findings suggest that intraarterial vasodilator therapy fails to improve intestinal viability after segmental mesenteric vascular occlusion. PMID- 11420491 TI - Intravascular extension of Wilms tumor. AB - OBJECTIVE: To define the incidence and manifestations of and optimal therapy for children with intravascular extension of Wilms tumor. METHODS: Children on a collaborative study of Wilms tumor who had intravascular extension into the inferior vena cava (IVC) or atrium were identified. Surgical checklists and surgical and pathology reports were reviewed. RESULTS: One hundred sixty-five of 2,731 patients had intravascular extension of Wilms tumor. The level of extension was IVC in 134 and atrium in 31. Sixty-nine had received preoperative therapy (55 with IVC extension and 14 with atrial extension) for a median of 8 weeks. Complications during preoperative chemotherapy were seen in five patients (tumor embolism and tumor progression in one each, and three with adult respiratory distress syndrome, one of which was fatal). The intravascular extension of the tumor regressed in 39 of 49 children with comparable pre- and posttherapy radiographic studies, including 7 of 12 in whom the tumor regressed from an atrial location, thus obviating the need for cardiopulmonary bypass. Surgical complications occurred in 36.7% of the children in the atrial group and 17.2% in the IVC group. The frequency of surgical complications was 26% in the primary resection group versus 13.2% in children with preoperative therapy. When all the complications of therapy were considered, including those that occurred during the interval of preoperative chemotherapy (one of the five also had a surgical complication), the incidence of complications among those receiving preoperative therapy was not statistically different from the incidence among those who underwent primary resection. The difference in 3-year relapse-free survival (76.9% for 165 patients with intravascular extension, 80.3% for 1,622 patients with no extension) was not statistically significant whether or not it was adjusted for stage and histology. CONCLUSIONS: Preoperative treatment of these children may facilitate resection by decreasing the extent of the tumor thrombus, but the overall frequency of complications is similar in both groups. PMID- 11420492 TI - Reoperation after recurrent groin hernia repair. AB - OBJECTIVE: To analyze reoperation rates for recurrent and primary groin hernia repair documented in the Swedish Hernia Register from 1996 to 1998, and to study variables associated with increased or decreased relative risks for reoperation after recurrent hernia. METHODS: Data were retrieved for all groin hernia repairs prospectively recorded in the Swedish Hernia register from 1996 to 1998. Actuarial analysis adjusted for patients' death was used for calculating the cumulative incidence of reoperation. Relative risk for reoperation was estimated using the Cox proportional hazards model. RESULTS: From 1996 to 1998, 17,985 groin hernia operations were recorded in the Swedish Hernia Register, 15% for recurrent hernia and 85% for primary hernia. At 24 months the risk for having had a reoperation was 4.6% after recurrent hernia repair and 1.7% after primary hernia repair. The relative risk for reoperation was significantly lower for laparoscopic methods and for anterior tension-free repair than for other techniques. Postoperative complications and direct hernia were associated with an increased relative risk for reoperation. Day-case surgery and local infiltration anesthesia were used less frequently for recurrent hernia than for primary hernia. CONCLUSIONS: Recurrent groin hernia still constitutes a significant quantitative problem for the surgical community. This study supports the use of mesh by laparoscopy or anterior tension-free repair for recurrent hernia operations. PMID- 11420493 TI - Re: Bowrey DJ et al. Gastroesophageal reflux disease in asthma. Effects of medical and surgical antireflux therapy on asthma control. Ann Surg 2000; 231:161 72. PMID- 11420494 TI - Re: Kuo PC et al. Laparoscopic donor nephrectomy with a 23-hour stay. A new standard for transplantation surgery. Ann Surg 2000; 231:772-779. PMID- 11420495 TI - Re: Stapfer M et al. Management of duodenal perforation after endoscopic retrograde cholangiopancreatography and sphincterotomy. Ann Surg 2000; 232:191-8. PMID- 11420496 TI - Re: Orozco et al. A comparative study of the elective treatment of variceal hemorrhage with beta-blockers, transendoscopic sclerotherapy and surgery. A prospective, controlled, and randomized trial during 10 years. Ann Surg 2000; 232:216-9. PMID- 11420497 TI - Tissue expansion using osmotically active hydrogel systems for direct closure of the donor defect of the radial forearm flap. AB - Although widely used, the radial forearm flap has been criticized for the poor quality of its donor site. Attempts to avoid donor-site problems have concentrated on the elaboration of the split-thickness and full-thickness skin graft methods of reconstruction. Skin grafts frequently fail over the flexor carpi radialis tendon, leading to chronic skin breakdown or, at best, tendon adhesion. Tissue expansion appears to be a good alternative that allows the use of local tissues to ultimately improve the forearm donor-site appearance. To avoid the disadvantages of traditional silicone balloon expanders (such as pressure peaks, infection, the valve at a distance from the expander, postoperative fillings), an osmotically active system was used. In an 18-month prospective study, 10 osmotically active hydrogel tissue expanders were placed on the forearms of 10 patients. The radial forearm flap was performed for intraoral reconstruction after surgical resection of oral cavity malignancies. The study showed that, in nine out of 10 patients, the expanded skin achieved was sufficient to cover the donor site after raising the forearm flap. Additionally, the expansion-related swelling pressure was well tolerated by the patients, the cosmetic results were very satisfactory, and the incidence of complications was very low. By using osmotically active hydrogel tissue expanders, there is no postoperative filling and no risk of complications arising from defective balloon expanders, filling valves, or missing ports. PMID- 11420499 TI - Eyelids and eye socket reconstruction using the expanded forehead flap and scapha composite grafting. AB - After trauma or excision of malignant tumor, it is difficult to achieve satisfactory results when reconstructing deformed eyelids and the socket for an ocular prosthesis. The authors demonstrate examples of successful reconstruction for a prosthetic eye that provided adequate and aesthetic soft-tissue support achieved by applying a three-step surgical procedure of reconstruction of the eye socket, the eyelids, and the tarsus and eyelid margin. Because it is highly vascularized and its distal end can be divided into two or three portions for easy three-dimensional reconstruction, the expanded forehead flap alone, with a galea flap, or with a free rectus abdominis muscle perforator flap was used. The expanded forehead flap also provides excellent thin upper lid contour and good color-matching with a recipient site. For the eye socket, sufficient volume of tissue was provided from the expanded forehead flap with or without a galea or a free rectus abdominis muscle perforator flap, and a deep and convex fornix was formed. This resulted in a good fit and in stability of the ocular prosthesis. The surface and the inner lining of the eyelids were reconstructed using portions of the expanded forehead flap. For the tarsus and eyelid margin, conventional reconstruction techniques use cartilage of the concha, which has limitations of length and which does not fit the shape of the tarsal margin. The authors used the scapha composite graft, and a natural shape and good elasticity resulted. PMID- 11420500 TI - Results with Furlow palatoplasty in management of velopharyngeal insufficiency. AB - A retrospective study was undertaken to assess speech outcomes in patients undergoing Furlow palatoplasty. Since 1994, the authors have used the position of the levator veli palatini musculature to determine type of surgical intervention recommended for the management of velopharyngeal insufficiency. Furlow palatoplasty has been used in patients with clinical evidence of sagittally oriented levator veli palatini musculature. Forty-eight patients who underwent a Furlow palatoplasty between June of 1994 and August of 1998 were included. All patients underwent preoperative and postoperative perceptual speech analyses to describe velopharyngeal insufficiency severity, nasal air emissions, and resonance, and preoperative nasendoscopy to assess velopharyngeal gap size and palatal and lateral pharyngeal wall movement. Other patient characteristics considered included gender, age at time of surgery, previously repaired cleft palate, submucous cleft palate, and syndrome diagnosis. Speech outcomes were determined on the basis of postoperative perceptual speech analyses and were categorized in one of three ways: (1) complete resolution of velopharyngeal insufficiency, (2) substantial improvement of velopharyngeal insufficiency, and (3) audible residual velopharyngeal insufficiency. Complete resolution of velopharyngeal insufficiency was defined as normal resonance and an absence of nasal air emissions. Substantial improvement of velopharyngeal insufficiency was defined as an improvement of at least two categories in velopharyngeal insufficiency severity in those patients without complete resolution. Audible residual velopharyngeal insufficiency refers to patients with postoperative velopharyngeal insufficiency severity ratings of mild, moderate, or severe. The male:female ratio in the study was 27:21. Twelve patients were syndromic; three had velocardiofacial syndrome. The median age at surgery was 6.5 years (range, 2 to 22 years). The average duration of follow-up was 14.7 months (range, 1.3 to 58.6 months). Postoperatively, the severity of velopharyngeal insufficiency was rated as none in 19 of the 48 patients (39.6 percent), minimal in eight (16.7 percent), mild in six (12.5 percent), moderate in nine (18.75 percent), and severe in six (12.5 percent). Substantial improvement was seen in seven of the 29 patients without complete resolution. There was a significant association between male gender and complete resolution of velopharyngeal insufficiency (p < 0.05). Presence of syndrome and female gender was associated with audible residual velopharyngeal insufficiency (p < 0.05). The main complication was palatal fistula (two cases). In conclusion, most patients who underwent a Furlow palatoplasty had a complete resolution or substantial improvement of velopharyngeal insufficiency postoperatively, and there were few surgical complications. PMID- 11420501 TI - The crumpled-ear deformity. AB - The common characteristics of the crumpled-ear deformity, the steps for surgical correction, and the authors' experience are described in this article. Commonly, the ear has a folded-over appearance; a normal length and width when unfurled; and wrinkling of the skin and cartilage of the helical rim, scapha, antihelix, and concha. Principles of correction of this deformity include (1) superficial scoring of the concave segments of the cartilage to open the wrinkled segments, (2) creating the antihelical fold by a mattress-suture technique, (3) repositioning of the helical rim, and (4) repositioning of the prominent ear lobe, if present. A total of 12 crumpled ears were evaluated in six patients, three of whom underwent surgery during their teenage years. No complications have been observed. A new classification of an ear deformity has been proposed based on morphology that to the authors' knowledge has not been described in the literature. Identification of the common features of the deformity allows for precise correction using already well-defined techniques. PMID- 11420502 TI - Treatment of mandible fractures using low-profile titanium miniplates: preliminary study. AB - This study evaluated the short-term results of patients treated with low-profile titanium miniplates for fractures of the mandible. Thirty-one fractures of the mandible in 23 patients were treated by open reduction and internal fixation using thin, low-profile miniplates and 1.3-mm self-threading screws. Duration of intermaxillary fixation ranged from 0 to 25 days. Patients were evaluated for complications during a follow-up period ranging from 6 to 24 months. Seven patients (30.4 percent) experienced complications. These included infection (n = 1), premature occlusal contact (n = 1), wound dehiscence (n = 1), temporomandibular joint disorder (n = 1), and paresthesia (n = 3). All complications were minor and adequately managed with incision and drainage, medication, and elastic traction. Low-profile titanium miniplates can be adequately used for internal fixation in selective mandibular fractures. Advantages of these types of plates include comfort due to the thinness of miniplates and ease of application. PMID- 11420503 TI - Preserved costal cartilage homograft application for the treatment of temporomandibular joint ankylosis. AB - Ankylosis of the temporomandibular joint has been a daunting problem in oral and maxillofacial surgery. Condylectomy with gap arthroplasty is the basic technique for treatment of the fully grown patient. In the past, reconstruction has primarily been accomplished with alloplastic materials or with autogenous tissue harvested from the patient. Joints reconstructed with alloplastic materials have been subject to complications such as acute infection and chronic inflammatory problems as a result of foreign-body reaction with the immune system. Biologic reconstruction with autogenous materials does expose the patient to the risk of complications at the donor site. In the last 4 years, we have treated seven patients between the ages of 20 and 42 years who had complete temporomandibular joint ankylosis. In each patient, the affected joint was exposed through an extended preauricular incision. The ankylosed mandibular condyle with the surrounding abnormal bone, together with the coronoid process, was resected and removed. The ankylosed area was resected until an improvement of at least 15 mm in the interincisal opening distance was obtained. A solvent-preserved homologous cartilage graft was sculpted according to the size and shape of the gap and was then placed in it as interpositional material. Physical therapy, including active and passive mouth-opening exercises, began on the second postoperative day and continued for 6 months. Patients were observed for 6 months to 4 years. During this period, no major complications were noted, and satisfactory results were obtained. The initial mean interincisal opening distance was 15.2 mm after surgery, and the final mean interincisal opening distance was 32 mm after completion of physiotherapy. No recurrence was seen during the 4 years of follow up. This technique seems to be an effective, time-saving, and simple alternative to other methods of joint reconstruction in adults who have fairly extensive ankylosis of the temporomandibular joint. In this article, a description of the surgical technique, a review of all cases, and recommendations for the use of this type of graft material are discussed. Our clinical experience over the past 4 years with the use of preserved homologous costal cartilage grafts as interpositional material has been encouraging. PMID- 11420504 TI - Scanning electron microscopy characterization of surgical instrument damage to breast implants. AB - In this article, mechanisms of breast-implant failure caused by surgical instruments commonly used to perform implantation, breast biopsies, needle localization procedures, cyst aspirations, and explantation are described. Failure was artificially induced in breast-implant shells using various types of surgical instruments, including scalpels, suture needles, hypodermic needles, hemostats, and Adson forceps. Field-emission scanning electron microscopy (SEM) was used to document the morphology of the failure sites produced by these instruments. Micrographs were used to categorize failure according to a specific type of surgical instrument. SEM micrographs were also obtained on explants that failed in situ, and the morphology of the corresponding failure sites was examined. The study was designed to document a range of failure mechanisms associated with gel-filled, saline-filled, double-lumen (saline-gel), and soybean oil-filled implants. The results of the study also demonstrate that SEM can often be used to determine the cause of breast-implant failure. PMID- 11420505 TI - Review of data describing outcomes that are used to assess changes in quality of life after reduction mammaplasty. AB - Although plastic surgeons have empirically "known" of the benefits of reduction mammaplasty for their patients, a paucity of outcome studies have been reported. For this study, an attempt to perform a meta-analysis of outcomes in reduction mammaplasty was undertaken. A computer literature search was performed of the MEDLINE database for the period between 1966 and September of 1997 for the Medical Subject Headings mammaplasty and outcome measures. Reference lists were used for additional reports. No trials were identified that met the criteria for meta-analysis. Seventeen publications met less restrictive review criteria that evaluated quality-of-life outcome measures. A systematic evaluation of patient focused outcome measures demonstrated that consistent improvement in physical symptoms was found across most studies, as was a high degree of patient satisfaction (78 to 95 percent very or moderately satisfied), and some have shown improvement in body image and psychological well-being. However, although this review does identify consistent improvements in patient quality of life after reduction mammaplasty, inconsistencies among study designs do not allow formal meta-analysis. PMID- 11420506 TI - Treatment of Bowen disease using the ultrasonic surgical aspirator. AB - In most cases, patients with Bowen disease lesions can be easily treated by surgical excision and direct closure; however, the ultrasonic surgical aspirator, an instrument in common use in many fields, offers a good treatment alternative with few drawbacks for patients with lesions that are large and/or located in joint areas, areas that are hard to heal, or areas that are surgically problematic such as the ears, nose, eyelids, and mucous membranes. The clinical application of the ultrasonic surgical aspirator in the treatment of Bowen disease promises considerable benefit in terms of patient comfort because removal of the lesion is easy, is noninvasive, and does not require hospitalization or result in restricted mobility. All of these factors, although benefiting patients, particularly the elderly, can also substantially lower the cost of treatment. PMID- 11420507 TI - How frequent is postmastectomy breast reconstructive surgery? A study linking two statewide databases. AB - Studies of the frequency of reconstructive surgery among breast cancer patients have involved non-population-based sources, incomplete ascertainment, or both. Postmastectomy breast reconstructive surgery among 4688 breast cancer patients (diagnosed between 1992 and 1996) was estimated by linking a population-based cancer registry with a statewide hospital discharge database in Connecticut. Of these 4688 patients, 585 (12.5 percent) had reconstruction coded in one database or both databases. The reconstruction rates were higher than in a previous study in Connecticut and increased from 9 percent in 1992 to 16 percent in 1996. Reconstruction was not related to the patient's tumor size, marital status, or race (black versus white), but declined with increasing age at diagnosis and with poverty rate (of the census tract of residence). These associations were similar to those reported from previous studies. PMID- 11420508 TI - Comparison of immediate and delayed free TRAM flap breast reconstruction in patients receiving postmastectomy radiation therapy. AB - Tumor pathologic features and the extent of nodal involvement dictate whether radiation therapy is given after mastectomy for breast cancer. It is generally well accepted that radiation negatively influences the outcome of implant-based breast reconstruction. However, the long-term effect of radiation therapy on the outcome of breast reconstruction with the free transverse rectus abdominis myocutaneous (TRAM) flap is still unclear. For patients who need postmastectomy radiation therapy, the optimal timing of TRAM flap reconstruction is controversial. This study compares the outcome of immediate and delayed free TRAM flap breast reconstruction in patients who received postmastectomy radiation therapy. All patients at The University of Texas M. D. Anderson Cancer Center who received postmastectomy radiation therapy and who also underwent free TRAM flap breast reconstruction between January of 1988 and December of 1998 were included in the study. Patients who received radiation therapy before delayed TRAM flap reconstruction were compared with patients who underwent immediate TRAM flap reconstruction before radiation therapy. Early and late complications were compared between the two groups. Early complications included vessel thrombosis, partial or total flap loss, mastectomy skin flap necrosis, and local wound healing problems, whereas late complications included fat necrosis, volume loss, and flap contracture of free TRAM breast mounds. Late complications were evaluated at least 1 year after the completion of radiation therapy for patients who had delayed reconstruction and at least 1 year after reconstruction for patients who had immediate reconstruction. During the study period, 32 patients had immediate TRAM flap reconstruction before radiation therapy and 70 patients had radiation therapy before TRAM flap reconstruction. Mean follow-up times for the immediate reconstruction and delayed reconstruction groups were 3 and 5 years, respectively. The mean radiation dose was 50 Gy in the immediate reconstruction group and 51 Gy in the delayed reconstruction group. One complete flap loss occurred in the delayed reconstruction group, and no flap loss occurred in the immediate reconstruction group. The incidence of early complications did not differ significantly between the two groups. However, the incidence of late complications was significantly higher in the immediate reconstruction group than in the delayed reconstruction group (87.5 percent versus 8.6 percent; p = 0.000). Nine patients (28 percent) in the immediate reconstruction group required an additional flap to correct the distorted contour from flap shrinkage and severe flap contraction. These findings indicate that, in patients who are candidates for free TRAM flap breast reconstruction and need postmastectomy radiation therapy, reconstruction should be delayed until radiation therapy is complete. PMID- 11420509 TI - Reconstructive surgery with a dermal regeneration template: clinical and histologic study. AB - Integra artificial skin was introduced in 1981 and its use in acute surgical management of burns is well established, but Integra has also been used in patients undergoing reconstructive surgery. Over a period of 25 months, the authors used Integra to cover 30 anatomic sites in 20 consecutive patients requiring reconstructive surgery and then analyzed the clinical and histologic outcomes. The most common reason for surgery was release of contracture followed by resurfacing of tight or painful scars. The authors assessed patients' satisfaction using a visual analog scale and scar appearance using a modified Vancouver Burn Index Scale. They evaluated the progress of wound healing by examining weekly punch-biopsy specimens with standard and immunohistochemical stains. Patients reported a 72 percent increase in range of movement, a 62 percent improvement in softness, and a 59 percent improvement in appearance compared with their preoperative states. Pruritus and dryness were the main complaints, and neither was improved much. Four distinct phases of dermal regeneration could be demonstrated histologically: imbibition, fibroblast migration, neovascularization, and remodeling and maturation. Full vascularization of the neodermis occurred at 4 weeks. The color of the wound reflected the state of neodermal vascularization. No adnexa, nerve endings, or elastic fibers were seen in any of the specimens. The new collagen was histologically indistinguishable from normal dermal collagen. The authors conclude that Integra is a useful tool in reconstructive surgery. The additional cost of its use can be justified by its distinct benefits compared with current methodology. PMID- 11420510 TI - Stimulated healing of recalcitrant wounds by topical application of enriched cell culture medium: a clinical report. AB - This study was designed to test the efficacy of enriched cell culture medium as a wound dressing. The rationale was to create within the wound space an optimal microenvironment, conducive to cellular proliferation, vascular granulation tissue formation, and epithelialization. This study was performed on various wounds that failed to respond to previous conventional treatments.A total of 288 wounds were within the inclusion criteria, with only contaminated and neoplastic wounds excluded. Most of the patients (80 percent) were ambulatory, and the wounds were examined by the attending physician once every 7 to 14 days at an outpatient clinic. The remaining 20 percent of patients were admitted to the study while hospitalized. Cell culture medium MCDB, supplemented with insulin, thyroxin, and growth hormone, was gelled. The gel was self-applied once a day to freshly washed wounds, covered with a gauze pad, and anchored with netting. Healing started 7 to 14 days after the initiation of treatment with enriched cell culture medium. However, the criterion for success of the treatment was determined on complete wound closure, which was achieved in 189 of 288 wounds (65.6 percent). Wound closure was correlated with the initial wound volume, stage, and origin. The average time required for closure of wounds caused by systemic pathologies (n = 181) and those based on regional status (n = 107) were 12.0 and 4.4 weeks, respectively, compared with 290 and 10.3 weeks of the previous conventional treatment. In 19 extensive wounds, when vascularized granulation tissue was established, a successful surgical closure was attained. Most wounds of patients who did not continue the enriched cell culture medium treatment (34.4 percent) manifested reduced wound volume, ranging from 11 to 98 percent of initial volume. Discontinuation of treatment was associated with difficulties in reaching the clinic for the weekly examination, rather than for reasons directly related to the treatment itself, and occurred significantly earlier during the treatment period.Thus, enriched cell culture medium was effective in stimulating wound healing in recalcitrant wounds. The healing was rapid with minimum scarring and pain. No side effects or allergic reactions were reported or observed. PMID- 11420511 TI - A comprehensive review of epinephrine in the finger: to do or not to do. AB - The prohibition against the use of local anesthetics with epinephrine for digital blocks or infiltration is an established surgical tradition. The present article provides a comprehensive review of all reported digital necrotic and ischemic complications with epinephrine in the digits in an effort to understand whether the current prohibition is based on documented reports. A comprehensive review of articles showing the successful use of local anesthetic with epinephrine in the digits is presented.A review of Index Medicus from 1880 to 1966 and a computer review of the National Library of Medicine database from 1966 to 2000 were performed using multiple keywords. Selected major textbooks from 1900 to 2000 were also reviewed.A total of 48 cases of digital gangrene after anesthetic blocks (mostly using cocaine or procaine) have been reported in the world literature. Only 21 cases involved the use of epinephrine; 17 involved an unknown concentration based on manual dilution. Multiple other concurrent conditions (hot soaks, tight tourniquets, and infection) existed in these case reports, making it difficult to determine the exact cause of the tissue insult. There have been no case reports of digital gangrene using commercial lidocaine with epinephrine (introduced in 1948). Multiple studies involving thousands of patients support the premise that the use of lidocaine with epinephrine is safe in the digits. An extensive literature review failed to provide consistent evidence that our current preparations of local anesthesia with epinephrine cause digital necrosis, although not all complications are necessarily reported. However, as with all techniques, caution is necessary to balance the risks of this technique with the dangers of mechanical tourniquets and upper extremity block anesthesia. PMID- 11420512 TI - Perforator-based flap in rats: a new experimental model. AB - A new type of flap, the perforator-based flap, has been described in the last decade. It has been used successfully as a pedicle or free flap by many plastic surgeons. There is no animal model for research, although these flaps have gained popularity in clinical use. We created a perforator-based flap model in the rat (a perforator-based flap group and two control groups; 10 rats in each group) and evaluated the survival characteristics of the new flap. The abdominal skin flap was elevated based on the second perforator of the right superior deep epigastric artery and then sutured to its original bed. In the first control group, the same flap was elevated with a subcutaneous pedicle without any perforator; in the second control group, a right-sided, random-pattern pedicle abdominal skin flap with the same dimensions and location was elevated and sutured to its original bed. Flap survival was studied, and microangiography and histologic studies were performed. The amount of viable skin in the three groups was compared 1 week later. The area of surviving skin paddles in the experimental group ranged from 74 to 83 percent; in the first control group, it was 0 percent; and in the second control group, it ranged from 29 to 44 percent (p < 0.001 and p < 0.001, respectively). There was a predictable and constant area of necrosis in the model. The results of this study demonstrate that most of the abdominal skin of the rat can survive on the basis of a single musculocutaneous perforator vessel. This flap can be easily elevated, and it can be used as a reliable model for flap research. PMID- 11420513 TI - Sensory restoration of the skin graft on a free muscle flap: experimental rabbit study. AB - Transplantation of a muscle flap with free skin graft for wound coverage is a common procedure in reconstructive microsurgery. However, the grafted skin has little or no sensation. Restoration of the sensibility of the grafted skin on the transferred muscle is critically important, especially in palmar hand, plantar foot, heel, and oral cavity reconstruction. The purpose of this study was to investigate the possibility of sensory restoration of the grafted skin on a trimmed muscle surface that has been sensory neurotized after sensory nerve-to motor nerve transfer, using the rabbit gracilis muscle as an animal model. The ipsilateral saphenous nerve (sensory) was transferred to the motor nerve of the gracilis muscle for sensory neurotization. A 4 x 4-cm2 area of skin island over the midportion of the gracilis muscle was harvested as a full-thickness skin graft. The upper half of the gracilis muscle was then excised, becoming a rough surface. The harvested skin was reapplied on the trimmed rough surface of the muscle. After 6 months, retrograde and antegrade horseradish peroxidase labeling studies were performed through skin and muscle injection. The group with a free skin graft was compared with the group with an intact surface of the gracilis muscle. This study clearly shows that sensory nerves can regenerate and penetrate into the trimmed muscle surface and grow into the overlying grafted skin. However, if the muscle surface is intact as with the compared group, sensory reinnervation of the grafted skin is not possible. PMID- 11420514 TI - Microsurgical tongue revascularization following oromandibular cancer resection. PMID- 11420515 TI - Unusual facial impalement injury. PMID- 11420516 TI - Closure of infratip nasal defect by two triangular flaps. PMID- 11420517 TI - Use of an extended glabellar fascial-cutaneous flap for reconstruction of the nasopharynx. PMID- 11420519 TI - Gracilis muscle free flap transfer using a radial artery/venae comitantes composite vascular pedicle. PMID- 11420520 TI - Venous superdrainage in deep inferior epigastric perforator flap breast reconstruction. PMID- 11420521 TI - Developments in craniomaxillofacial surgery: use of self-reinforced bioabsorbable osteofixation devices. AB - Because of the problems associated with the conventional osteofixation devices used in craniomaxillofacial surgery, absorbable devices present an appealing alternative. Devices made of the polymers polylactide, polyglycolide, and their copolymers (PLGA and P[L/DL]LA) are currently the most commonly used. Ultrahigh strength implants can be manufactured from these polymers with the self reinforcing technique. Over the authors' almost two decades of study, both in experimental and clinical settings, self-reinforced devices have proved to be biocompatible, easy to handle, and mechanically strong, even for the fixation of femoral neck fractures. In craniomaxillofacial surgery, the authors have used self-reinforced devices for over 8 years without complications. Because of the more favored degradation characteristics, currently the copolymeric self reinforced devices (P[L/DL]LA, Biosorb FX and PLGA, Biosorb PDX; Elite Performance Technologies, Solana Beach, Calif.) represent the advancing front in the application of absorbable devices in craniomaxillofacial surgery. The authors' share their experience and their studies of self-reinforced devices, which possess the highest strength and ductility of all bioabsorbable products. PMID- 11420522 TI - Primary repair of bilateral cleft lip and nasal deformity. AB - LEARNING OBJECTIVES: After studying this article, the participant should be able to: 1. List five principles that guide synchronous repair of bilateral complete cleft lip and nasal deformity. 2. Explain how different growth rates for the principal nasolabial features are applied during primary repair. 3. Describe two approaches for positioning the alar cartilages to form the columella. 4. Discuss the influences on referral patterns for a newborn with bilateral cleft lip. - Traditional repair of bilateral cleft lip focused on labial closure but accentuated the nasal deformities, which were addressed later. By the end of the past century, single-staged labial closure had replaced the old multistaged procedures and the technical emphasis had begun to shift from secondary to primary nasal correction. Now, presurgical maxillary orthopedics sets the bony foundation for synchronous nasolabial repair and for closure of the alveolar clefts. The study of normal nasolabial growth and the typical stigmata of the conventional methods provides the necessary foreknowledge to guide surgical sculpture in three dimensions and to anticipate the fourth dimension. The convergence of several forces are changing referral lines for children born with bilateral cleft lip. These include affirmation of centers of excellence, surgeons' self-regulation, prenatal diagnosis, economics of health-care delivery, and increasing parental sophistication. These pressures are not necessarily in conflict. Care by a subspecialized plastic surgeon and experienced team is in the best interests of the child and the third-party payer. PMID- 11420523 TI - Common traits for thinking out-of-the-box: the evolution of cosmetic surgery. PMID- 11420524 TI - Endoscopically assisted, intraorally approached corrective rhinoplasty. AB - In the field of facial surgery, operations that require guesswork can result in unexpected complications. One example of such "blind" facial surgery is the lateral osteotomy procedure in corrective rhinoplasty. In most conventional corrective rhinoplasties, the postoperative results of a lateral osteotomy can be controlled by the surgeon's visual perception or manual dexterity; therefore, an experienced surgeon is indispensable in such elaborate operations. Until now, reports have focused on the endoscopic approach through the nasal dorsum or septum through the nostril. However, because of the difficulty in handling the endoscope with osteotomy instruments, it is considered difficult to perform a precise lateral osteotomy procedure using that approach. The authors think the intraoral endoscopic approach should be considered a viable alternative in corrective rhinoplasty. Through small, bilateral gingivobuccal incisions, both the piriform apertures and nasal bones can be easily exposed, and the exact level of the lateral osteotomy can be confirmed directly under the endoscope. The lateral osteotomy is made simply with a reciprocating saw, and symmetrical cutting can be ascertained during the operation. Sometimes, a particular osteotomy level or the proper repositioning of osteotomed segments can be readily evaluated with assistance from the endoscope during the operation. Eleven cases using this procedure were performed over the past 3 years. These endoscopic repairs for a deviated nose were quite helpful for visual confirmation and accurate correction. No complications occurred when using the endoscope with this procedure. PMID- 11420526 TI - Treatment guidelines for botulinum toxin type A for the periocular region and a report on partial upper lip ptosis following injections to the lateral canthal rhytids. AB - Inactivation of the orbicularis oculi muscle by chemodenervation with botulinum toxin type A (Botox, Allergan, Inc., Irvine, Calif.) as a sole procedure or in conjunction with blepharoplasty has proved to be a reliable method to improve the appearance of the periocular area. Botox has the unique and ideal characteristic in that, with repeated use, there is potential for a prolonged clinical effect with smaller dosages. In addition, if a complication does arise--while not aesthetically acceptable and potentially untoward--it is time-limited, and the anatomical area will eventually return to its pretreatment baseline status. In this study, in three cases [in more than 1000 crow's feet treatment sessions (2000 sides)] over the course of 1 year, partial lip ptosis resulting from weakening of the zygomaticus major muscle after the injection of Botox into the periocular region are reported. This article reviews suggested treatment guidelines and anatomic considerations for the periocular region to maintain injection standardization and improve the safety profile of Botox as the aesthetic indications for its use expand and the number of individuals who inject it increases. PMID- 11420528 TI - Surgical correction of the face with the square jaw and weak chin: angle-to-chin bone transfer. PMID- 11420530 TI - Analysis of the effects of deep mechanical massage in the porcine model. AB - Deep mechanical massage has been advocated as an alternative or adjunctive therapy for the contouring of subcutaneous fat and as a treatment for cellulite. We evaluated the effects of deep mechanical massage using two pig models. Yucatan pigs were divided into three groups (n = 4). One side of each body received 4, 10, or 20 treatments and the other side served as a control. Full-thickness tissue sections, including the underlying muscle, were harvested from identical treated and untreated regions. Examination of these regionally matched samples revealed an accumulation of dense, longitudinal collagen bands in the middle dermal and deep subdermal regions, which progressively increased with the number of treatments. Distortion and disruption of adipocytes was noted. In Yorkshire pigs, force-transducing balloon catheters were surgically placed between the deep subcutaneous tissue and muscle fascia. Catheters were inserted into two regions with different skin and subcutaneous tissue characteristics, the midflank and the hip. Standardized maneuvers were performed at suction settings 3, 5, 7, and 9 to record baseline tissue forces. Each maneuver carried a unique force signature. The measurement of tissue forces was repeated on the opposite side after 10 standardized treatment sessions. Analysis showed a significant reduction of measured forces at the midflank after the treatments. The actual force measured with each particular maneuver varied between different operators but not with different suction settings, suggesting that the technique of administering the treatments is the primary factor in creating the force within the tissue. This leads to the conclusion that deep mechanical massage is highly dependent on the individual operator of the device. PMID- 11420531 TI - Blood loss in major liposuction procedures: a comparison study using suction assisted versus ultrasonically assisted lipoplasty. AB - The blood loss that accompanies liposuction procedures has always been a concern. Tumescent injection of the targeted area of liposuction with dilute lidocaine and epinephrine solution has minimized intraoperative blood loss. Proponents of a newer ultrasonically assisted lipoplasty technique have claimed many benefits over traditional suction-assisted lipoplasty. However, few quantitative data are available on the intraoperative blood loss and the significance of postoperative anemia using the ultrasonic method. A prospective clinical observational design was used to investigate 38 patients undergoing suction-assisted lipoplasty and 37 patients undergoing ultrasound-assisted lipoplasty in whom the liposuction aspirate was expected to be more than 1000 ml. These patients were investigated with preoperative measurement of hemoglobin, platelet count, prothrombin time, partial thromboplastin time, and postoperative measurement of hemoglobin on the seventh postoperative day. In addition, hemoglobin concentration and whole blood volume were calculated from the infranatant portion of the liposuction aspirate. The mean +/- SD volume of the liposuction aspirate was 2901 +/- 1471 ml for suction-assisted compared with 2741 +/- 1086 ml for ultrasound-assisted lipoplasty. The mean +/- SD of whole blood volume in liposuction aspirate per case was 36 +/- 50.82 ml for suction-assisted lipoplasty and 36 +/- 28.62 ml for ultrasound-assisted lipoplasty. The mean +/- SD of the preoperative hemoglobin concentration was 13.93 +/- 0.99 g/dl for suction-assisted lipoplasty and 14.05 +/- 1.16 g/dl for ultrasound-assisted lipoplasty, whereas the mean +/- SD of the postoperative hemoglobin concentration was 13 +/- 1.42 g/dl for suction-assisted lipoplasty and 13.05 +/- 1.32 g/dl for ultrasound-assisted lipoplasty. The mean decrease in hemoglobin on the seventh postoperative day was 0.93 +/- 0.92 g/dl for suction-assisted lipoplasty and 1 +/- 0.64 g/dl for ultrasound-assisted lipoplasty. The volume of whole blood loss was estimated to be 12.4 ml in each 1000 ml of liposuction aspirate when using suction-assisted lipoplasty versus 13.1 ml when using ultrasound-assisted lipoplasty. All procedures were done under general anesthesia, and patients were discharged home on the same day. No blood transfusion was required. This study shows that blood loss using the ultrasonic technique is slightly higher, though insignificant, than when using suction. However, this study did not demonstrate a difference in the postoperative hemoglobin decrease between the two techniques. PMID- 11420533 TI - Port-wine gingivo-alveolar enlargement: the solution. PMID- 11420534 TI - Standards for cosmetic surgery articles. PMID- 11420535 TI - The theory of exclusivity. PMID- 11420536 TI - Standards for cosmetic surgery articles. PMID- 11420537 TI - An adjunct to substernal dissection in esophageal reconstruction. PMID- 11420538 TI - Is there an etiological link between the computer and clefts of the lip? PMID- 11420540 TI - Rejuvenation of the midface. PMID- 11420539 TI - A simple time-saving technique for nasal taping after rhinoplasty. PMID- 11420541 TI - Cleft earlobes in males. PMID- 11420542 TI - The "tie-over dressing" refined. PMID- 11420543 TI - Has your finger ever been scratched by suture? PMID- 11420544 TI - Auricular cartilage scoring and correcting a flattened antihelix. PMID- 11420545 TI - Massive prepatellar bursa: a case of natural tissue expansion: anatomic and histologic implications. PMID- 11420546 TI - Repositioning of a drain. PMID- 11420547 TI - Fibrous ring "pulls plug" on saline-filled implant. PMID- 11420548 TI - Using Dermabond. PMID- 11420549 TI - A wound after contact between a child and a dog does not necessarily mean an animal bite. PMID- 11420550 TI - Botox for contraction of pectoral muscles. PMID- 11420551 TI - Patients having removal of silicone breast implants. PMID- 11420552 TI - The nasal bone graft for nasal augmentation. PMID- 11420553 TI - Plastic surgeon's responsibility following breast reconstruction. PMID- 11420563 TI - [Metallic elements in tissues surrounding internal rigid fixation (IRF) devices]. AB - BACKGROUND: Plates and other devices made by several alloys have been introduced to reach the stability of bone fractured fragments. Elements constituting alloys could be detected especially in organs, yet also in local tissues. Aim of the present study is the analysis of tissues surrounding IRF devices analyzing the morphology of released particles and studying the behavior of adjacent tissues to check metallic elements diffusion. METHODS: Biopsies were retrieved from 18 patients, aged 20 to 76 years. The patients received IRF by plates, screws and grids from 4 months to 9 years. They were divided into five groups according to the local phlogistic degree. Ordinary light microscopy, scanning electron microscopy and X-ray microprobe analysis (EDS system) was used to perform morphological investigations and identification of metal particles and elements. RESULTS: Metal particles or elements arising from plates, screws or grid may undergo tissular diffusion and cellular uptake. Not only Chromium, Iron or Aluminium but also Titanium may be easily released from devices and engulfed in tissues. In particular Titanium diffusion is evident in fibrous tissue surrounding IRF devices. Aluminium appears to be particularly accumulated in a persistent way in fibrous tissues and shows a characteristic embedding pattern in lamellar bone. CONCLUSIONS: The degree of local phlogosis appears to be strictly correlated to metallosis. Chromium, Iron, Aluminium and also Titanium, even if at different degree, give rise to phlogistic effects. Metallosis and phlogosis can produce a cascade process in which they are both the cause and the effect at the same time. The abundant release of Titanium, which does not normally produce clinical phlogosis as i.e. Aluminium, should be worthy of further investigations on its cellular effects. PMID- 11420564 TI - [Opinions and attitudes on oral cancer in a sample of students attending a state secondary school in Rome. A pilot study]. AB - BACKGROUND: This study aimed to evaluate knowledge and awareness in a group of secondary school students on the subject of risk factors and strategies used in the prevention of oral cancer. METHODS: The study took the form of a questionnaire which was filled in by a group of 106 secondary school students. The questionnaire was divided into 3 sections: the first analysed the general characteristics of the sample population; the second investigated their knowledge on the specific subject of this study; the last regarded exposure to risk factors and the role played by the dentist and family doctor in the prevention of oral cancer. RESULTS: Of the 106 students taking part in the study, 42% were male and 58% female. 30% of the group felt it was not possible to prevent cancer in general. 6% associated cigarette smoking with oral cancer, whereas 15% identified alcohol as a risk factor for the same pathology. 30% of the group thought that it was not possible to prevent oral cancer. In the event of a suspected oral lesion, 44% would consult the family doctor, 25% would go to the dentist and 3% to a dental technician. CONCLUSIONS: This study highlights the need improve knowledge of prevention methods in oral cancer. Although young people do not have a high risk of developing oral cancer, they represent a means of conveying information in a social and familial context, thus enabling an early diagnosis, and they also represent the preferred target for primary prevention activities. PMID- 11420565 TI - [Biology of root resorption process in deciduous teeth. Review of the literature]. AB - Aim of this paper, is to examine, with a review of the literature the different aspects of physiological process of root resorption of deciduous teeth. The process of development of mouth and teeth and their structure is also described. Studies made during the last period, in most part by Japanese authors, improved our knowledge about the process of root resorption of deciduous teeth, even if many aspects are still unknown. The involvement of the pulp and the periodontal ligament and the presence of enzimatic activity and immunologic cells have been confirmed; all these elements seem to have a definite role in the process of root resorption of the deciduous teeth. PMID- 11420566 TI - [Circulating biomarkers association in the follow-up of patients with oral cancer]. AB - The goal of this study is to analyze the importance of circulating biomarkers association in the management of patients affected by oral cancer. In this study a survey is made of the international experience from 1980 to 1990 based on the presence of CEA, LASA, SCC Ag, TPA, ferritina, CA-50 and others in patients affected by oral cancer and the sensitivity and specificity of these circulating biomarkers association are assessed. In patients with active disease, the results obtained at the time of diagnosis of oral cancer are not satisfactory due to poor specificity of these circulating biomarkers association. The conclusions is drawn that the circulating biomarkers association (especially CEA, SCC Ag, LASA, ferritin, TPA and CA-50) appears to be useful in the prognosis and staging of oral cancer, while their presence is not significative for the diagnosis. PMID- 11420567 TI - [Extracorporeal shock wave lithotripsy in the therapy of salivary calculi: efficacy, risks and limits. Review of the literature. Note 1]. AB - The authors carry out a review of the international literature on an alternative methodology to surgical therapy of lithiasis of the salivary glands, indicating information, limitations and possible complications. More than fifty papers by international authors were analysed regarding their experience on the use of electro-magnetic waves in the treatment of salivary gland lithiasis. It was observed that extracorporeal lithotripsy is an alternative and valid method to surgery, considering that the complications are negligible. Considering the expectations and the relationship between cost and advantage, extracorporeal lithotripsy can be considered as first treatment with surgery as second therapeutic choice. PMID- 11420568 TI - [Morphofunctional aspects of the stomatognathic apparatus in the totally edentulous patient. Prosthetic rehabilitation]. AB - The authors examined the morphofunctional aspects of the various structures of the stomatognathic apparatus in the totally edentulous patient with the aim of carrying out and incorporating full prosthetic restoration. The following aspects are evaluated: 1) the reactive behaviour or resilience of the fibromucous support in the various areas under the functional load and its functional conditioning; 2) the vertical and horizontal intermaxillary ratios. Its modifications in an edentulous subject; 3) the evaluation and optimisation of the verticentre, namely the correct stable functional centre reference; 4) the regulation of muscle strength in relation to the length of contraction of muscle fibre; 5) type 3 lever mechanism in mandibular function and the creation of a balance distribution of occlusal forces in the centre. A suitable functional occlusive scheme in the total prosthesis achieves a correct adjustment of muscle contraddiction and a balanced distribution of load forces on the supporting tissues and on the temporomandibular joints. It constitutes a functional centre reference which centralises and stabilises the jaw and the prosthesis. In addition to correct technical installation, this also depends on a detailed preliminary diagnostic evaluation of the anatomofunctional conditions of all the structures in the stomatognathic apparatus of the subject to be treated. PMID- 11420569 TI - [Scanning electron microscopy and light microscopy under polarized light of a submandibular salivary gland calculus]. AB - BACKGROUND: The salivary calculus is an ancient pathologic condition, called sublingual stone by Hyppocrates. It is well-defined from a anatomo-pathologic, diagnostic and topographic viewpoint, though some pathogenesis-related doubts still exist. This work aims at relating the micro-morpho-structural aspect of a salivary calculus of remarkable dimensions with the current calculogenetic hypotheses. METHODS: A 1.1 g salivary calculus removed from the initial section of Wrthon s duct in the right-hand submandibular gland of a 42 year old male has been studied. Following the fixation in 4% buffered formalin and the inclusion in resin, it was serially sectioned in 15-20 micron slices. Some sections were coloured with toluidine blue O and acid fuchsin. The exeresis of the calculus was carried out intraorally, with marsupialization under local anesthesia. The analysis was performed using a SEM with backscattered electrons and a polarised and transmitted light microscope. RESULTS: The investigations have outlined the presence of various organic cores and a concentric, stratiform architecture interrupted by radial elements. CONCLUSIONS: The formation of this calculus may be due to mixed secondary lithiasis resulting from the formation of colloids and crystalloids. PMID- 11420570 TI - The selective serotonin reuptake inhibitor sertraline: its profile and use in psychiatric disorders. AB - The naphthylamine derivative sertraline is a potent and selective inhibitor of serotonin reuptake into presynaptic terminals. Sertraline has a linear pharmacokinetic profile and a half-life of about 26 h. Its major metabolite, desmethylsertraline does not appear to inhibit serotonin reuptake. Sertraline mildly inhibits the CYP2D6 isoform of the cytochrome P450 system but has little effect on CYP1A2, CYP3A3/4, CYP2C9, or CYP2C19. It is, however, highly protein bound and may alter blood levels of other highly protein bound agents. Sertraline is a widely used serotonin reuptake inhibitor that has been shown to have both antidepressant and antianxiety effects. Many clinical trials have demonstrated its efficacy in depression compared with both placebo and other antidepressant drugs. Its efficacy has also been demonstrated in randomized, controlled trials of patients with obsessive-compulsive disorder, panic disorder, social phobia, and premenstrual dysphoric disorder. In short-term, open-label studies it has appeared efficacious and tolerable in children and adolescents and in the elderly, and data are positive for its use in pregnant or lactating women. Typical side effects include gastrointestinal and central nervous system effects as well as treatment-emergent sexual dysfunction; withdrawal reactions may be associated with abrupt discontinuation of the agent. The safety profile of sertraline in overdose is very favorable. Sertraline's efficacy for both mood and anxiety disorders, relatively weak effect on the cytochrome P450 system, and tolerability profile and safety in overdose are factors that contribute to make it a first-line agent for treatment in both primary and tertiary care settings. PMID- 11420571 TI - Paroxetine: a review. AB - Paroxetine is a potent and selective serotonin reuptake inhibitor (SSRI) with currently approved indications for the treatment of depression, obsessive compulsive disorder, panic disorder and social phobia. It is also used in the treatment of generalized anxiety disorder, post traumatic stress disorder, premenstrual dysphoric disorder and chronic headache. Paroxetine, a phenylpiperidine derivative, is the most potent inhibitor of the reuptake of serotonin (5-hydroxytryptamine, 5-HT) of all the currently available antidepressants including the class of SSRIs. It is a very weak inhibitor of norepinephrine (NE) uptake but it is still more potent at this site than the other SSRIs. The selectivity of paroxetine, i.e., the ratio of inhibition of uptake of norepinephrine to serotonin (NE/5-HT) is amongst the highest of the SSRIs. Paroxetine has little affinity for catecholaminergic, dopaminergic or histaminergic systems and by comparison with tricyclic antidepressants (TCAs) has, therefore, a reduced propensity to cause central and autonomic side effects. Paroxetine exhibits some affinity for the muscarinic cholinergic receptor but much less than the TCAs. In addition, the adaptive changes of somatodendritic (5 HT(1A)) and terminal (5-HT(1B/1D)) autoreceptors observed with paroxetine are different to those observed with TCAs; it also inhibits nitric oxide synthase. It is both a substrate and an inhibitor of cytochrome isoenzyme P450 2D6. Paroxetine is well absorbed orally and undergoes extensive first pass metabolism that is partially saturable. Its metabolites are pharmacologically inactive in vivo. Steady state levels are achieved after 4-14 days and an elimination half-life of 21 h is consistent with once-daily dosing. There is wide inter-individual variation in the pharmacokinetics of paroxetine in adults as well as in the young and the elderly with higher plasma concentrations and slower elimination noted in the latter. Elimination is also reduced in severe renal and hepatic impairment. Serious adverse events are, however, extremely rare even in overdose. In summary, paroxetine is well tolerated and effective in the treatment of both depressive and anxiety disorders across the age range. PMID- 11420572 TI - Sevoflurance: approaching the ideal inhalational anesthetic. a pharmacologic, pharmacoeconomic, and clinical review. AB - Sevoflurane is a safe and versatile inhalational anesthetic compared with currently available agents. Sevoflurane is useful in adults and children for both induction and maintenance of anesthesia in inpatient and outpatient surgery. Of all currently used anesthetics, the physical, pharmacodynamic, and pharmacokinetic properties of sevoflurane come closest to that of the ideal anesthetic (200). These characteristics include inherent stability, low flammability, non-pungent odor, lack of irritation to airway passages, low blood:gas solubility allowing rapid induction of and emergence from anesthesia, minimal cardiovascular and respiratory side effects, minimal end-organ effects, minimal effect on cerebral blood flow, low reactivity with other drugs, and a vapor pressure and boiling point that enables delivery using standard vaporization techniques. As a result, sevoflurane has become one of the most widely used agents in its class. PMID- 11420573 TI - International conferences: international opportunities. PMID- 11420574 TI - Framework Convention Alliance: building support for global tobacco control. PMID- 11420575 TI - To the extreme. PMID- 11420576 TI - Electrocardiographic findings in athletes: the prevalence of left ventricular hypertrophy and conduction defects. AB - OBJECTIVES: To determine whether there are electrocardiographic differences or distinctive abnormalities between athletes and sedentary subjects, and to verify the relationship between vagal activity measured by heart rate variability (SD of all normal-to-normal intervals [SDNN]) and possible electrocardiographic abnormalities. SUBJECTS AND METHODS: Resting electrocardiograms and heart rate variability measurements were performed separately during a single visit on 100 athletes and 50 nonathlete control subjects aged 18 to 55 years. The athletes were from the following various sports disciplines: long-distance running, mountain biking, cross-country skiing, biathlon, speed skating, swimming and triathlon. RESULTS AND CONCLUSIONS: There were significantly longer RR intervals, PR intervals and QT intervals in athletes than in control subjects (all P<0.05). The QRS complex and QTc did not show significant differences (both P>0.05). The prevalence of left ventricular hypertrophy (LVH) and incomplete right bundle branch block (IRBBB) was 10% and 7%, respectively, in athletes, but these conditions were absent in control subjects; among athletes, 2% presented with both conditions. LVH and IRBBB were more common among long-distance runners (six of 14 and four of 14, respectively) and could be attributed to normal, long term adaptation to intense, repeated exercise. LVH was related to age (P=0.04), whereas IRBBB was influenced by the number of years of training in the respective sports discipline (P=0.03). The mean SDNN value was significantly more elevated in athletes (P=0.0001), reflecting a higher parasympathetic tone than in sedentary control subjects. However, there was no relationship between vagal activity and LVH or IRBBB (both P>0.05). PMID- 11420577 TI - The apolipoprotein E genotype influences the risk for vasospastic angina. AB - It has been suggested that people with the epsilon4 allele of the apolipoprotein E (apoE) polymorphism and the deletion (D) allele of the insertion (I/D) polymorphism of angiotensin-converting enzymes, are at a greater risk for coronary artery disease. However, only a few studies have examined the relationships between vasospastic angina (VSA) and genotype, especially with the apoE polymorphism. In the present study, 76 patients with VSA without significant fixed coronary artery stenosis, 149 patients with ischemic heart disease (IHD) who had 75% or more luminal diameter narrowing and 213 healthy subjects were enrolled. The odds ratio for VSA of the epsilon4 allele carriers relative to the epsilon3/3 allele subjects compared with subjects with IHD and control subjects combined was 0.44 (95% CI 0.21 to 0.93, P=0.021), and that compared with control subjects alone was 0.36 (95% CI 0.17 to 0.78, P=0.005), implying that the presence of the epsilon4 allele indicates resistance to the development of VSA. In contrast, people with the epsilon2 allele showed a tendency to develop VSA more frequently than did patients with IHD (P=0.009), although the frequency of the epsilon2 allele did not differ between patients with VSA and control subjects. On the other hand, no recessive and dominant effects of the D alleles on VSA were found. These findings suggest that the risk of the occurrence of VSA may be reduced by the epsilon4 allele and increased by the epsilon2 allele. The ApoE polymorphism may be associated with IHD and VSA, probably due to the modulation of lipid metabolism. PMID- 11420578 TI - Differential coronary artery calcification detected by electron beam computed tomography as an indicator of coronary stenosis among patients with stable angina pectoris. AB - BACKGROUND: The detection of coronary artery calcification by electron beam computed tomography (EBCT) has been suggested as an indicator of atherosclerosis and coronary artery disease (CAD). There is no consensus on the correlation between coronary calcification and angiographically significant stenosis on an artery-by-artery basis. OBJECTIVE: To examine the relationship between coronary calcification score (CCS) and the presence of significant CAD on an artery-by artery basis in patients with stable angina pectoris. METHODS AND RESULTS: EBCT and coronary angiogram (CAG) were evaluated in 71 patients with stable angina and in nine control subjects. The CCSs of each of the four major coronary arteries were highest in patients with significant CAD (n=43), followed by patients with insignificant CAD (n=5), patients with syndrome X (n=23) and control subjects, respectively. Calcification scores of the four major coronary arteries appeared to have different predictive power for significant stenosis on the same vessel. For left main (LM) and left anterior descending (LAD) coronary arteries, CCSs of vessels with significant stenoses were not different from those without significant stenoses (values expressed as medians: LM 0 versus 1; LAD 98.5 versus 70; not significant). Calcification scores of left circumflex (LCX) and right coronary arteries (RCA) were significantly higher in vessels with significant stenosis (LCX 49.5 versus 0; RCA 53 versus 1; P<0.05). CCSs appeared to be moderately useful to predict significant stenoses in these two vessels (areas under receiver operating characteristic curves: LCX 0.68+/-0.08, 95% CI 0.52 to 0.81; RCA 0.71+/-0.08, 95% CI 0.55 to 0.84). CONCLUSIONS: The CCSs of RCA and LCX arteries, but not those of LM and LAD arteries, may predict significant angiographic stenosis on an artery-by-artery basis among patients with stable angina pectoris. PMID- 11420579 TI - Rationale and design of the Canadian Continuous Positive Airway Pressure Trial for Congestive Heart Failure patients with Central Sleep Apnea--CANPAP. AB - BACKGROUND: Cheyne-Stokes respiration with central sleep apnea (CSR-CSA) is commonly observed in patients with congestive heart failure (CHF), in which it is an independent risk factor for death. Treating CSR-CSA may, therefore, improve outcomes in patients with CHF. OBJECTIVE: The Canadian Continuous Positive Airway Pressure Trial for Congestive Heart Failure Patients with Central Sleep Apnea (CANPAP) is a multicentre, randomized, controlled clinical trial designed to test the hypothesis that treating CSR-CSA with continuous positive airway pressure (CPAP) will reduce the combined rate of all-cause mortality and cardiac transplantation in patients with CHF. Secondary outcomes include the severity of CSR-CSA, left ventricular volumes and function, submaximal exercise capacity, quality of life and hospital admissions. PATIENTS AND METHODS: The aim is to enroll 408 patients with CHF (New York Heart Association class II to IV and left ventricular ejection fraction of less than 40%) and CSR-CSA over a five-year period. Patients are randomly assigned to either standard medical therapy for CHF or standard medical therapy plus CPAP. The trial has 80% power to detect a 35% between-group treatment difference for the all-cause mortality-cardiac transplantation rate. Substudies will assess the role of oximetry as a screening tool for detecting CSR-CSA in patients with CHF, and the effects of CPAP on cardiac arrhythmias and plasma concentrations of natriuretic peptides and catecholamines. CONCLUSIONS: The CANPAP trial will help to define the role of CPAP better as a nonpharmacological intervention for the treatment of patients with CHF who have CSR-CSA. PMID- 11420580 TI - Impaired left ventricular systolic function after a half-ironman race. AB - OBJECTIVE: To assess the effects of prolonged strenuous exercise in the form of a half-ironman (HI) race (2 km swim, 90 km bike ride and a 21 km run) on left ventricular systolic function. METHODS AND RESULTS: The study participants consisted of nine male triathletes (mean age +/- SD 32+/-5 years) who competed in the Great White North HI race. Two-dimensional transthoracic echocardiograms were obtained two to three days before the HI (prerace), immediately after completion of the HI (postrace) and 24 h after cessation of exercise. Compared with before the race, performing an HI was associated with a decline in systolic blood pressure (prerace 127.2+/-15.0 mmHg compared with after the race 116.1+/-10.2 mmHg, P<0.05), the systolic blood pressure to end-systolic cavity area ratio (a surrogate for left ventricular contractility - prerace 14.3+/-3.0 mmHg/cm(2) compared with postrace 11.0+/-2.2 mmHg/cm(2), P<0.05) and the fractional area change (prerace 54.1+/-3.8% compared with postrace 47.4+/-5.5%, P<0.05). There was also a concomitant increase in heart rate (prerace 56.3+/-9.4 beats/min compared with postrace 74.1+/-10.7 beats/min, P<0.05), the end-systolic cavity area (prerace 9.2+/-2.2 cm(2) compared with postrace 10.8+/-1.9 cm(2), P<0.05) and the end-systolic cavity area to end-systolic myocardial area ratio (prerace 0.39+/-0.08 compared with postrace 0.51+/-0.1, P<0.05), which returned toward baseline values 24 h after cessation of the HI. CONCLUSIONS: Performing an HI appears to be associated with a transient impairment in left ventricular contractility and a subsequent decline in left ventricular systolic function that tends to return toward normal values within 24 h after cessation of exercise. PMID- 11420581 TI - The evaluation of physician patterns of practice in cardiac patients at two British Columbia tertiary referral hospitals. AB - OBJECTIVE: To evaluate the patterns of care in patients hospitalized with acute coronary syndrome or congestive heart failure. DESIGN: A pilot study - retrospective chart review. SETTING: Two British Columbia tertiary referral teaching hospitals. PATIENTS: One hundred patients admitted with stable angina, acute ischemic syndrome or congestive heart failure at each hospital. RESULTS: Patient profiles showed that women at both hospitals were significantly older and had significantly longer lengths of stay than men. Patterns of practice were comparable except when it came to lipid testing and lipid-lowering agents being prescribed to patients at the time of discharge. At hospital A, patients were more likely to have a lipid profile performed compared with patients at hospital B. CONCLUSIONS: The addition of lipid testing to the clinical pathway for patients with myocardial infarction led to more patients at hospital A having a lipid profile performed. The routine assessment of lipid profiles in patients admitted for acute ischemic syndromes led to more aggressive pharmacological treatment with hydroxy-methyl-glutaryl coenzyme A reductase inhibitors - agents that are known to reduce mortality and recurrent infarction. Methods to improve treatment are proposed to help reduce morbidity and/or mortality in patients with ischemic heart disease and congestive heart failure. PMID- 11420582 TI - Lest we forget: Early Canadian contributions to the care of children with congenital heart malformations. AB - In a century of remarkable progress in medicine, one of the outstanding stories is the development of successful treatment of congenital heart malformations. This achievement is the outcome of the ideas and research of many people. Many of the early 'transforming' advances originated in Canada. Maude Abbott catalogued and classified heart malformations in a clinically meaningful manner. Arthur Charles and David Scott in Toronto, Ontario, produced a clinically useful heparin preparation, and the studies of Bill Bigelow led to the application of hypothermia in cardiac surgery. John Keith and Bill Mustard at Toronto's Hospital for Sick Children, and Arnold Johnson and Tony Dobell at the Montreal Children's Hospital, Montreal, Quebec, established the first Canadian programs devoted to the correction of congenital heart defects in childhood. Mustard devised the first widely successful operation for transposition of the great arteries. Flavio Coceani and Peter Olley discovered the role of prostaglandin E in the ductus arteriosus, and applied that knowledge clinically. The turn of the century is an appropriate time to celebrate these Canadian successes. PMID- 11420583 TI - The beneficial effects of plant sterols on serum cholesterol. AB - Phytosterol-enriched margarines are a recent addition to the list of so-called 'functional foods'. The ingestion of phyto-sterols lowers the serum cholesterol by inhibiting intestinal uptake of the sterol. The phytosterols available in consumer products are comprised predominantly of beta-sitosterol and sitostanol. The esterified form of these phytosterols increases their solubility and enhances their residence time in the small intestine. Their ability to displace cholesterol from micelles in the small intestine underlies the mechanism that inhibits cholesterol absorption, leading to a 10% reduction in total serum cholesterol. Numerous well designed studies have documented the beneficial actions of these phytosterols on serum cholesterol. PMID- 11420584 TI - Papillary muscle rupture complicating an acute myocardial infarction. PMID- 11420585 TI - More on health care. PMID- 11420586 TI - Canadian guidelines for cardiac rehabilitation and atherosclerotic heart disease prevention: a summary. AB - Atherosclerotic heart disease (AHD) is the leading cause of death in Canadian women and men. Cardiac rehabilitation has been repeatedly shown to reduce cardiac morbidity and mortality significantly among patients with documented AHD. The Canadian Association of Cardiac Rehabilitation (CACR) has defined cardiac rehabilitation as "the enhancement and maintenance of cardiovascular health through individualized programs designed to optimize physical, psychological, social, vocational and emotional status. This process includes the facilitation and delivery of secondary prevention through heart hazard (risk factor) identification and modification in an effort to prevent disease progression and the recurrence of cardiac events". This summary presents a limited amount of background information and the majority of clinical practice recommendations contained within the previously published CACR Guidelines. These evidence-based clinical recommendations are intended as guidelines to good clinical practice rather than as standards of care. The key focus of this summary is the need for complete and targeted intervention of all heart hazards in patients at high or very high risk for, or with documented, AHD. To achieve this goal, the CACR Guidelines and this summary present risk stratification strategies designed to determine unambiguously a patient's risk of exercise-related cardiac events (short term absolute risk or disease prognosis) and their risk of recurrent AHD events (long term absolute risk from disease progression). The establishment of the short term and long term absolute AHD risks can then be used to determine heart hazard targets and the type of exercise program prescribed for patients with AHD. Despite the use of evidence-based medical practices, none of the recommendations presented in this document can replace the expert judgment of properly trained and experienced cardiac rehabilitation professionals. Health care providers must always be free to choose where and when clinical practice guidelines are applied, modified or superceded, depending on individual patient circumstances. PMID- 11420587 TI - Too late, again? PMID- 11420588 TI - The changing medical research scene. PMID- 11420589 TI - The epidemiology of childhood asthma in Red Deer and Medicine Hat, Alberta. AB - OBJECTIVES: To document the prevalence of asthma among school-aged children in two Alberta communities, to understand host and indoor environmental factors associated with asthma, and to compare these factors between the two communities. DESIGN: A cross-sectional study with a nested, case-control follow-up. SETTING: Red Deer and Medicine Hat, Alberta. PATIENTS AND METHODS: Questionnaires were sent to families of children aged five to 19 years in Red Deer (n=5292) and Medicine Hat (n=5372) to identify children with current asthma. A random sample of 592 children with current asthma and 443 with no history of asthma constituted a case-control population; they were followed up by telephone to obtain responses to the European Respiratory Health Survey and, in children with current asthma, the Pediatric Quality of Life Questionnaire. RESULTS: Cross-sectional response rates were 84% and 73% for Red Deer and Medicine Hat, respectively. The prevalence of asthma was higher in Medicine Hat (17.0%) than in Red Deer (12.8%). In the follow-up study, factors associated with the presence of asthma were parental asthma or allergies, number of siblings, presence of cats, serious respiratory illnesses before five years of age, sex, age, presence of mould and/or mildew and use of a gas cooking stove. The presence of mould and/or mildew was a significant risk factor in Red Deer but not in Medicine Hat. CONCLUSIONS: Asthma prevalence among school children in Red Deer was consistent with recently published Canadian data; the prevalence in Medicine Hat was higher than expected, especially given the low relative humidity. Risk factor data are consistent with other studies in that parental asthma, especially maternal asthma, was a significant predictor of childhood asthma. Cats in the house (both communities) and environmental tobacco smoke (Medicine Hat only) were significantly less common among children with asthma, suggesting that preventive actions may have been taken in the homes of some children with asthma. PMID- 11420590 TI - Comparison of intramuscular betamethasone and oral prednisone in the prevention of relapse of acute asthma. AB - OBJECTIVE: To compare the relapse rate after a single intramuscular injection of a long acting corticosteroid, betamethasone, with oral prednisone in patients discharged from the emergency department (ED) for acute exacerbations of asthma. PATIENTS AND METHODS: Patients with acute exacerbations of asthma who were suitable for discharge from the ED were enrolled in a double-blind, randomized, placebo controlled pilot study. At discharge, patients were randomly assigned to receive either intramuscular betamethasone 12 mg and placebo capsules, or a placebo intramuscular injection and prednisone 50 mg daily for seven days. At days 7 and 21, patients were contacted by telephone to determine relapse. Relapse was defined as an unscheduled visit to a physician for treatment of continuing or worsening symptoms of asthma. RESULTS: One hundred and seventy-one patients were enrolled, of whom 87 were randomly assigned to the betamethasone group and 84 to the prednisone group. Baseline characteristics were matched evenly between the groups, with the exception of asthma duration (15.5 versus 21.2 years, respectively) and use of inhaled corticosteroids (46% versus 64.3% respectively) (P<0.05). Using intention-to-treat analysis, the relapse rates for betamethasone and prednisone at day 7 were 14.9% (13 of 87 patients) and 25% (21 of 84 patients), respectively (P=0.1), and at day 21, the rates were 36.8% (32 of 87 patients) and 31% (26 of 84 patients), respectively (P=0.4). There were no differences in symptom score, peak flows and adverse effects between the two groups at days 7 and 21. CONCLUSIONS: A single dose of intramuscular betamethasone 12 mg was safe and as efficacious as prednisone in preventing the relapse of acute asthma. There was a trend toward a reduced relapse rate at seven days. In select ED patients discharged for acute asthma, intramuscular betamethasone may be an effective alternative to prednisone. PMID- 11420591 TI - Underdiagnosis of chronic obstructive pulmonary disease: a rationale for spirometry as a screening tool. AB - Chronic obstructive pulmonary disease (COPD) is associated with significant morbidity and mortality. It is the fourth leading cause of death in the United States, and its impact on quality of life can be severe. The debate on spirometry as a screening tool has been raised and dropped on various occasions over the past 30 years. This paper readdresses the debate in the light of recent evidence from population studies. Spirometry is an underused, easy to perform office-based procedure that has been further facilitated by the advent of modern technology. Despite the fact that spirometry is the gold standard for the diagnosis and assessment of COPD, mass screening using this tool remains controversial. This article provides a discussion based on a recent review of the literature regarding the current and future status of spirometry as a screening tool. A thoughtful approach to spirometry screening should include assessments of the magnitude of underdiagnosis, potential effectiveness of intervention, predictive value of spirometry and clinical profile of patients with COPD. PMID- 11420592 TI - Clinical practice guidelines for suctioning the airway of the intubated and nonintubated patient. AB - OBJECTIVE: To provide physicians, physiotherapists, nurses and respiratory therapists with guidelines for the application of airway suctioning. DESIGN: This clinical practice guideline was developed using the model by Browman and colleagues. A working group of representatives from four professional colleges (nurses, physicians and surgeons, physiotherapists and respiratory therapists) and research experts was formed to conduct a systematic review, develop evidence based recommendations and generate clinical practice guidelines. MEDLINE (1966 to 1998), CINAHL (1982 to 1997) and EMBASE (1974 to 1996) as well as the reference lists of identified articles were searched. Inclusion of articles was determined by at least two group members, and studies were classified according to type. Randomized, controlled; randomized; and nonrandomized crossover and comparative cohort trials were grouped by type of intervention and population for use in the development of recommendations. Other observational and animal studies dealing with adverse effects of suctioning were included in the review but were not used in the development of recommendations. Input on the evidence-based recommendations was sought and incorporated from members of all four professions and from experts on content and methodology. SETTING: Any setting (hospital or home) where suctioning is performed. POPULATION: Intubated and nonintubated adults, infants and children. RESULTS AND CONCLUSIONS: An attempt was made to develop recommendations in each of the subcategories of suctioning techniques addressed by at least one study. In some subcategories, definite recommendations were made (13 in adults, and three in children and infants); in other subcategories, insufficient evidence precluded recommendations. The recommendations addressed the following aspects of suctioning: preoxygenation, hyperinflation, insufflation, hyperoxygenation, hyperventilation, saline instillation, adaptor use, medication use, open and closed systems, and various types of catheters. PMID- 11420593 TI - Cystic angiomatosis of the bone appearing as intrathoracic lung masses. AB - A rare case of cystic angiomatosis of the bone (CAB) manifesting as bilateral intrathoracic masses is described. In 1996, a 47-year-old woman was referred to the chest clinic at St Boniface Hospital for evaluation of chronic cough, and her chest radiograph showed large, bilateral pleural-based intrathoracic masses. Ten years before, she had been diagnosed with CAB following bone biopsies of her hip and knee. Computed tomography revealed two chest wall masses with internal septations of bone. CAB is a rare, multicentric disease characterized by involvement of the vascular and lymphatic systems, producing diffuse cystic lesions in the skeleton. Although the ribs are frequently involved, the presentation with intrathoracic masses is uncommon. PMID- 11420594 TI - Kiss and run mechanism in exocytosis. PMID- 11420595 TI - Modeling the current-voltage characteristics of charophyte membranes. II. The effect of salinity on membranes of Lamprothamnium papulosum. AB - Lamprothamnium is a salt-tolerant charophyte that inhabits a broad range of saline environments. The electrical characteristics of Lamprothamnium cell membranes were modeled in environments of different salinity: full seawater (SW), 0.5 SW, 0.4 SW, and 0.2 SW. The cells were voltage-clamped to obtain the I/V (current-voltage) and G/V (conductance-voltage) profiles of the cell membranes. Cells growing at the different salinities exhibited one of three types of I/V profiles (states): pump-, background- and K(+)-states. This study concentrates on the pump- and background-states. Curved (pump-dominated) I/V characteristics were found in cells with resting membrane PDs (potential differences) of -219 +/- 12 mV (in 0.2 SW: 6 cells, 16 profiles), -161 +/- 12 mV (in 0.4 SW: 6 cells, 7 profiles), -151 +/- 12 mV (in 0.5 SW: 6 cells, 12 profiles) and -137 +/- 12 mV (in full SW: 8 cells, 13 profiles). The linear I/V characteristics of the background-state were found in cells with resting PDs of -107 +/- 12 mV (in 0.4 SW: 7 cells, 12 profiles), -108 +/- 12 mV (in 0.5 SW: 7 cells, 10 profiles) and 104 +/- 12 mV (in full SW: 3 cells, 5 profiles). The resting conductance (G) of the cells progressively increased with salinity, from 0.5 S x m(-2) (in 0.2 SW) to 22.0 S x m(-2) (in full SW). The pump peak conductance only rose from 2 S x m( 2) (0.2 SW) to 5 S x m(-2) (full SW), accounting for the increasingly depolarized resting PD observed in cells in more saline media. Upon exposure to hypertonic medium, both the pump and an inward K+ rectifier were stimulated. The modeling of the I/V profiles identified the inward K+ rectifier as an early electrical response to hypertonic challenge. PMID- 11420596 TI - IP3-gated channels and their occurrence relative to CNG channels in the soma and dendritic knob of rat olfactory receptor neurons. AB - Olfactory receptor neurons respond to odorants with G protein-mediated increases in the concentrations of cyclic adenosine 3',5'-monophosphate (cAMP) and/or inositol-1,4,5-trisphosphate (IP3). This study provides evidence that both second messengers can directly activate distinct ion channels in excised inside-out patches from the dendritic knob and soma membrane of rat olfactory receptor neurons (ORNs). The IP3-gated channels in the dendritic knob and soma membranes could be classified into two types, with conductances of 40 +/- 7 pS (n = 5) and 14 +/- 3 pS (n = 4), with the former having longer open dwell times. Estimated values of the densities of both channels from the same inside-out membrane patches were very much smaller for IP3-gated than for CNG channels. For example, in the dendritic knob membrane there were about 1000 CNG channels x microm(-2) compared to about 85 IP3-gated channels x microm(-2). Furthermore, only about 36% of the dendritic knob patches responded to IP3, whereas 83% of the same patches responded to cAMP. In the soma, both channel densities were lower, with the CNG channel density again being larger ( approximately 57 channels x microm(-2)) than that of the IP3-gated channels ( approximately 13 channels x microm(-2)), with again a much smaller fraction of patches responding to IP3 than to cAMP. These results were consistent with other evidence suggesting that the cAMP-pathway dominates the IP3 pathway in mammalian olfactory transduction. PMID- 11420597 TI - Hydrogen peroxide inhibits gap junctional coupling and modulates intracellular free calcium in cochlear Hensen cells. AB - The double whole-cell patch-clamp configuration was applied to analyze gap junctional conductance (Gj) of isolated pairs of cochlear supporting Hensen cells of guinea pig under control conditions and in the presence of hydrogen peroxide (H2O2). Under control conditions, the dependence of Gj on transjunctional voltage (Vj) appeared to vary between different cell pairs with a maximum value of about 40 nS at Vj close to 0 mV. The voltage dependence and the maximum amplitude of Gj stayed constant for at least 2 hr. Addition of H2O2 to the bath at concentrations above 0.08 mm caused a significant decrease of Gj, but the membrane potential of about -30 mV was not affected. In parallel, intracellular free calcium ([Ca2+]i) was followed using fura-2. At 0.8 mm H2O2, a sustained increase of [Ca2+]i was observed, while 0.08 mm H2O2 evoked an oscillating-like behavior of [Ca2+]i. We propose that the H2O2-evoked inhibition of gap junctional coupling of Hensen cells is closely related to pathophysiological conditions such as noise- induced hearing loss, aminoglycoside-related ototoxicity and presbycusis, which are known to be associated with production of free radicals. PMID- 11420598 TI - Single water channels of aquaporin-1 do not obey the Kedem-Katchalsky equations. AB - The Kedem-Katchalsky (KK) equations are often used to obtain information about the osmotic properties and conductance of channels to water. Using human red cell membranes, in which the osmotic flow is dominated by Aquaporin-1, we show here that compared to NaCl the reflexion coefficient of the channel for methylurea, when corrected for solute volume exchange and for the water permeability of the lipid membrane, is 0.54. The channels are impermeable to these two solutes which would seem to rule out flow interaction and require a reflexion coefficient close to 1.0 for both. Thus, two solutes can give very different osmotic flow rates through a semi-permeable pore, a result at variance with both classical theory and the KK formulation. The use of KK equations to analyze osmotic volume changes, which results in a single hybrid reflexion coefficient for each solute, may explain the discrepancy in the literature between such results and those where the equations have not been employed. Osmotic reflexion coefficients substantially different from 1.0 cannot be ascribed to the participation of other 'hidden' parallel aqueous channels consistently with known properties of the membrane. Furthermore, we show that this difference cannot be due to second-order effects, such as a solute-specific interaction with water in only part of the channel, because the osmosis is linear with driving force down to zero solute concentration, a finding which also rules out the involvement of unstirred-layer effects. Reflexion coefficients smaller than 1.0 do not necessitate water-solute flow interaction in permeable aqueous channels; rather, the osmotic behaviour of impermeable molecular-sized pores can be explained by differences in the fundamental nature of water flow in regions either accessible or inaccessible to solute, created by a varying cross-section of the channel. PMID- 11420599 TI - Inhibition of glial Na+ and K+ currents by tamoxifen. AB - Tamoxifen (tmx) is a non-steroidal triphenylethylene derivative that is predominantly known as a competitive antagonist at the estrogen receptor and is used in the treatment of breast cancer. Recent studies suggest that tamoxifen is also beneficial in the treatment of brain metastases and primary brain tumors. Tmx accumulates in brain and its concentration can be up to 46-fold higher than in serum. Therefore, astrocytes may be exposed to tmx in vivo. We use the whole cell patch-clamp technique to examine the effects of tmx on voltage-dependent cation currents in rat cortical cultures. Using biophysical and pharmacological methods, we isolate sustained and transient outward potassium currents (I(KS) and I(KT), respectively), inwardly rectifying potassium currents (I(KIR)), and transient inward sodium currents (I(Na)). We show that that TTX-sensitive I(Na) is completely inhibited by 10 microm tmx within 5 min. Similarly, tmx blocks I(KS), but does not inhibit I(KT) or I(KIR) at these concentrations. Tmx effects are irreversible with 10 min wash. Interestingly, the currents sensitive to tmx are important in growth control of glial cells (MacFarlane & Sontheimer, 1997). Therefore, we examine cytotoxic and proliferative effects of tmx. Tmx (10 microm) is not cytotoxic as judged by trypan blue exclusion. However, incubation with tmx significantly reduces cell proliferation as examined by 3[H]-thymidine uptake. PMID- 11420600 TI - Intracellular domains of mouse connexin26 and -30 affect diffusional and electrical properties of gap junction channels. AB - To evaluate the influence of intracellular domains of connexin (Cx) on channel transfer properties, we analyzed mouse connexin (Cx) Cx26 and Cx30, which show the most similar amino acid sequence identities within the family of gap junction proteins. These connexin genes are tightly linked on mouse chromosome 14. Functional studies were performed on transfected HeLa cells stably expressing both mouse connexins. When we examined homotypic intercellular transfer of microinjected neurobiotin and Lucifer yellow, we found that gap junctions in Cx30 transfected cells, in contrast to Cx26 cells, were impermeable to Lucifer yellow. Furthermore, we observed heterotypic transfer of neurobiotin between Cx30 transfectants and HeLa cells expressing mouse Cx30.3, Cx40, Cx43 or Cx45, but not between Cx26 transfectants and HeLa cells of the latter group. The main differences in amino acid sequence between Cx26 and Cx30 are located in the presumptive cytoplasmic loop and C-terminal region of these integral membrane proteins. By exchanging one or both of these domains, using PCR-based mutagenesis, we constructed Cx26/30 chimeric cDNAs, which were also expressed in HeLa cells after transfection. Homotypic intercellular transfer of injected Lucifer yellow was observed exclusively with those chimeric constructs that coded for both cytoplasmic domains of Cx26 in the Cx30 backbone polypeptide chain. In contrast, cells transfected with a construct that coded for the Cx26 backbone with the Cx30 cytoplasmic loop and C-terminal region did not show transfer of Lucifer yellow. Thus, Lucifer yellow transfer can be conferred onto chimeric Cx30 channels by exchanging the cytoplasmic loop and the C-terminal region of these connexins. In turn, the cytoplasmic loop and C-terminal domain of Cx30 prevent Lucifer yellow transfer when swapped with the corresponding domains of Cx26. In chimeric Cx30/Cx26 channels where the cytoplasmic loop and C-terminal domains had been exchanged, the unitary channel conductance was intermediate between those of the parental channels. Moreover, the voltage sensitivity was slightly reduced. This suggests that these cytoplasmic domains interfere directly or indirectly with the diffusivity, the conductance and voltage gating of the channels. PMID- 11420602 TI - The involvement of sphingolipids in multidrug resistance. AB - Administration of most chemotherapeutic agents eventually results in the onset of apoptosis, despite the agents' variety in structure and molecular targets. Ceramide, the central molecule in cellular glycosphingolipid metabolism, has recently been identified as an important mediator of this process. Indeed, one of the events elicited by application of many cytotoxic drugs is an accumulation of this lipid. Treatment failure in cancer chemotherapy is largely attributable to multidrug resistance, in which tumor cells are typically cross-resistant to multiple chemotherapeutic agents. Different cellular mechanisms underlying this phenomenon have been described. Of these the drug efflux pump activity of P glycoprotein and the multidrug resistance-associated proteins are the most extensively studied examples. Recently, an increased cellular capacity for ceramide glycosylation has been recognized as a novel multidrug resistance mechanism. Indeed, virtually all multidrug-resistant cells exhibit a deviating sphingolipid composition, most typically, increased levels of glucosylceramide. On the other hand, several direct molecular interactions between sphingolipids and drug efflux proteins have been described. Therefore, in addition to a role in the multidrug resistance phenotype by which ceramide accumulation and, thus, the onset of apoptosis are prevented, an indirect role for sphingolipids might be envisaged, by which the activity of these efflux proteins is modulated. In this review, we present an overview of the current understanding of the interesting relations that exist between sphingolipid metabolism and multidrug resistance. PMID- 11420603 TI - Sodium blocking induced by a point mutation at the C-terminal end of the pore helix of the KAT1 channel. AB - A plant hyperpolarization-activating K+ channel, KAT1, is highly selective for K+ over Na+ and is little affected by external Na+, which is crucial to take up K+ effectively in a Na+-containing environment. It has been shown that a mutation at the location (Thr256) preceding the selectivity signature sequence dramatically enhanced the sensitivity of the KAT1 channel to external Na+. We report here electrophysiological experiments for the mechanism of action of external Na+ on KAT1 channels. The Thr256 residue was substituted with either glutamine (Q) or glutamate (E). The wild-type channel was insensitive to external Na+. However, the activity of both mutant channels was significantly depressed by Na+ with apparent dissociation constants of 6.7 mm and 11.3 mm for T256Q and T256E, respectively. The instantaneous current-voltage relationships revealed distinct blocking mechanisms for these mutants. For T256Q a typical voltage-dependent fast blocking was shown. On the other hand, the blocking for the T256E mutant was voltage-independent at low Na+ concentrations and became voltage-dependent at higher concentrations. At extreme hyperpolarization the blocking was relieved significantly. These data strongly suggest that the mutation at the end of the pore helix rearranged the selectivity filter and allows Na+ to penetrate into the pore. PMID- 11420604 TI - GABA concentration sets the conductance of delayed GABAA channels in outside-out patches from rat hippocampal neurons. AB - GABAA channels were activated by GABA in outside-out patches from rat cultured hippocampal neurons. They were blocked by bicuculline and potentiated by diazepam. In 109 of 190 outside-out patches, no channels were active before exposure to GABA (silent patches). The other 81 patches showed spontaneous channel activity. In patches containing spontaneous channel activity, rapid application of GABA rapidly activated channels. In 93 of the silent patches, channels could be activated by GABA but only after a delay that was sometimes as long as 10 minutes. The maximum channel conductance of the channels activated after a delay increased with GABA concentration from less than 10 pS (0.5 microm GABA) to more than 100 pS (10 mm GABA). Fitting the data with a Hill-type equation gave an EC50 value of 33 microm and a Hill coefficient of 0.6. The channels showed outward rectification and were chloride selective. In the presence of 1 microm diazepam, the GABA EC50 decreased to 0.2 microm but the maximum conductance was unchanged. Diazepam decreased the average latency for channel opening. Bicuculline, a GABA antagonist, caused a concentration-dependent decrease in channel conductance. In channels activated with 100 microm GABA the bicuculline IC50 was 19 microm. The effect of GABA on channel conductance shows that the role of the ligand in GABAA receptor channel function is more complex than previously thought. PMID- 11420605 TI - Distinct mechanisms of [Ca2+]i oscillations in HSY and HSG cells: role of Ca2+ influx and internal Ca2+ store recycling. AB - This study examined [Ca2+]i oscillations in the human salivary gland cell lines, HSY and HSG. Relatively low concentrations of carbachol (CCh) induced oscillatory, and higher [CCh] induced sustained, steady-state increases in [Ca2+]i and KCa currents in both cell types. Low IP3, but not thapsigargin (Tg), induced [Ca2+]i oscillations, whereas Tg blocked CCh-stimulated [Ca2+]i oscillations in both cell types. Unlike in HSG cells, removal of extracellular Ca2+ from HSY cells (i) did not affect CCh-stimulated [Ca2+]i oscillations or internal Ca2+ store refill, and (ii) converted high [CCh]-induced steady-state increase in [Ca2+]i into oscillations. CCh- or thapsigargin-induced Ca2+ influx was higher in HSY, than in HSG, cells. Importantly, HSY cells displayed relatively higher levels of sarcoendoplasmic reticulum Ca2+ pump (SERCA) and inositoltrisphosphate receptors (IP3Rs) than HSG cells. These data demonstrate that [Ca2+]i oscillations in both HSY and HSG cells are primarily determined by the uptake of Ca2+ from, and release of Ca2+ into, the cytosol by the SERCA and IP3R activities, respectively. In HSY cells, Ca2+ influx does not acutely contribute to this process, although it determines the steady-state increase in [Ca2+]i. In HSG cells, [Ca2+]i oscillations directly depend on Ca2+ influx; Ca2+ coming into the cell is rapidly taken up into the store and then released into the cytosol. We suggest that the differences in the mechanism of [Ca2+]i oscillations HSY and HSG cells is related to their respective abilities to recycle internal Ca2+ stores. PMID- 11420606 TI - Adrenergic regulation of ion transport across adult alveolar epithelial cells: effects on Cl- channel activation and transport function in cultures with an apical air interface. AB - The effect of beta-adrenergic receptor stimulation on Cl- channel activation was investigated in alveolar epithelial cells grown in monolayer culture and in freshly isolated cells. Monolayers cultured under apical air interface conditions exhibited enhanced amiloride-sensitive Na+ transport compared to apical liquid interface monolayers. Amiloride or benzamil inhibited most (66%) of the basal short circuit current (Isc) with half-maximal inhibitory concentration (IC50) values of 0.62 microm and 0.09 microm respectively. Basolateral addition of terbutaline (2 microm) produced a rapid decrease in Isc followed by a slow recovery that exceeded the basal Isc. When Cl- was replaced with methanesulfonate in either intact monolayers or basolateral membrane permeabilized monolayers, the response to terbutaline (2 microm) was completely inhibited. No effect of terbutaline on amiloride-sensitive Na+ current was detected. beta-Adrenergic agonists and 8-chlorothiophenyl cyclic adenosine monophosphate (8-ctp cAMP) directly stimulated a Cl- channel in freshly isolated alveolar epithelial cells. The current was blocked by glibenclamide (100 microm) and had a reversal potential of -22 mV. No increase in amiloride-sensitve current was detected in response to terbutaline or 8-cpt cAMP stimulation. These data support the conclusion that beta-adrenergic agonists produce acute activation of apical Cl- channels and that monolayers maintained under apical air interface conditions exhibit increased Na+ absorption. PMID- 11420607 TI - Osmotic stimulation of the Na+/H+ exchanger NHE1: relationship to the activation of three MAPK pathways. AB - The Na+/H+ exchanger (NHE) becomes activated by hyperosmolar stress, thereby contributing to cell volume regulation. The signaling pathway(s) responsible for the shrinkage-induced activation of NHE, however, remain unknown. A family of mitogen-activated protein kinases (MAPK), encompassing p42/p44 Erk, p38 MAPK and SAPK, has been implicated in a variety of cellular responses to changes in osmolarity. We therefore investigated whether these kinases similarly signal the hyperosmotic activation of NHE. The time course and osmolyte concentration dependence of hypertonic activation of NHE and of the three sub-families of MAPK were compared in U937 cells. The temporal course and dependence on osmolarity of Erk and p38 MAPK activation were found to be similar to that of NHE stimulation. However, while pretreatment of U937 cells with the kinase inhibitors PD98059 and SB203580 abrogated the osmotic activation of Erk and p38 MAPK, respectively, it did not prevent the associated stimulation of NHE. Thus, Erk1/2 and/or p38 MAPK are unlikely to mediate the osmotic regulation of NHE. The kinetics of NHE activation by hyperosmolarity appeared to precede SAPK activation. In addition, hyperosmotic activation of NHE persisted in mouse embryonic fibroblasts lacking SEK1/MKK4, an upstream activator of SAPK. Moreover, shrinkage-induced activation of NHE still occurred in COS-7 cells that were transiently transfected with a dominant-negative form of SEK1/MKK4 (SEK1/MKK4-A/L) that is expected to inhibit other isoforms of SEK as well. Together, these results demonstrate that the stimulation of NHE and the activation of Erk, p38 MAPK and SAPK are parallel but independent events. PMID- 11420608 TI - A triple mutant, K319N/H322Q/E325Q, of the lactose permease cotransports H+ with thiodigalactoside. AB - In a previous study, we characterized a lactose permease mutant (K319N/E325Q) that can transport H+ ions with sugar. This result was surprising because other studies had suggested that Glu-325 plays an essential role in H+ binding. To determine if the lactose permease contains one or more auxiliary H+ binding sites, we began with the K319N/E325Q strain, which catalyzes a sugar-dependent H+ leak, and isolated third site suppressor mutations that blocked the H+ leak. Three types of suppressors were obtained: H322Y, H322R, and M299I. These mutations blocked the H+ leak and elevated the apparent Km value for lactose. The M299I and H322Y suppressors could still transport H+ with beta-d thiodigalactoside (TDG), but the H322R strain appeared uncoupled for H+/sugar cotransport. Four mutant strains containing a nonionizable substitution at codon 322 (H322Q) were analyzed. None of these were able to catalyze uphill accumulation of lactose, however, all showed some level of substrate-induced proton accumulation. The level seemed to vary based on the substrate being analyzed (lactose or TDG). Most interestingly, a triple mutant, K319N/H322Q/E325Q, catalyzed robust H+ transport with TDG. These novel results suggest an alternative mechanism of lactose permease cation binding and transport, possibly involving hydronium ion (H3O+). PMID- 11420609 TI - Molecular basis of pituitary dysfunction in mouse and human. PMID- 11420610 TI - Fine mapping of Ath6, a quantitative trait locus for atherosclerosis in mice. AB - Ath6 is a novel quantitative trait locus associated with differences in susceptibility to atherosclerosis between C57BL/6J (B6) and C57BLKS/J (BKS) inbred mouse strains. Combining data from an intercross and a backcross (1593 meioses) between mice from B6 and BKS strains and from The Jackson Laboratory interspecific backcross panels, (C57BL/6J x Mus spretus) F1 x C57BL/6J and (C57BL/6J x SPRET/Ei) F1 x SPRET/Ei, we constructed a consensus genetic map and narrowed Ath6 to a 1.07 +/- 0.26 cM interval between the anonymous DNA marker D12Pgn4 and the gene Nmyc1. This region is near the proximal end of murine Chromosome (Chr) 12, which is homologous to the human chromosomal region 2p24 p25. Marker order in the Ath6 region was concordant among the two crosses and The Jackson Laboratory interspecific backcross panels. This high resolution map rules out candidate genes encoding apolipoprotein B, syndecan 1, and Adam17. The two Ath6 crosses have a combined potential resolution of 0.06 cM. PMID- 11420611 TI - Quantitative trait loci affecting the behavior of A/J and CBA/J intercross mice in the elevated plus maze. AB - How allelic diversity affects neural mechanisms to produce behavioral variation is largely unknown. The elevated plus maze, consisting of open and closed arms, has been used as a model of behavioral variation in rodent exploration. Under dim illumination the nature of the sensory stimuli that influence arm choice is uncertain. Two inbred mouse strains, A/J (Tyrc/Tyrc, the albino phenotype, mutation in tyrosinase) with a strong preference for closed arm entry, and CBA/J (Pdebrdl/Pdebrdl, the retinal degeneration phenotype, mutation in the beta subunit of rod cGMP phosphodiesterase), with a weak preference for open arm entry, were studied under varying light. Because behavioral differences persist under red light, variation in light perception is not likely to fully account for variation in arm choice. To identify genetic factors influencing arm choice (100 x Open arm entries/Total arm entries) quantitative trait loci analyses (QTL) were performed on (A/J x CBA/J)F2 mice. Two QTLs, one of which includes PDEB, were identified on Chr 5 (LOD > 10) and account for > 30% of the behavioral variation in arm preference. Tyr (Chr 7, 44 cM) was linked to closed arm entries but not arm preference, and is unlikely to be acting through a direct effect on light perception, because A/J arm entries were not affected by red light and there was no interaction with PDEB in the (A/J x CBA/J)F2 mice. Whether the candidate QTLs on Chr 5 affect arm choice through an effect on light perception is unknown, but phenotypic differences between F2 mice with retinal degeneration and CBA/J mice and F2 mice with albinism and A/J mice suggest that factors other than light sensitivity contribute to arm preference in these two strains. PMID- 11420612 TI - Comparative physical mapping of targeted regions of the rat genome. AB - The comparative mapping and sequencing of vertebrate genomes is now a key priority for the Human Genome Project. In addition to finishing the human genome sequence and generating a 'working draft' of the mouse genome sequence, significant attention is rapidly turning to the analysis of other model organisms, such as the laboratory rat (Rattus norvegicus). As a complement to genome-wide mapping and sequencing efforts, it is often important to generate detailed maps and sequence data for specific regions of interest. Using an adaptation of our previously described approach for constructing mouse comparative and physical maps, we have established a general strategy for targeted mapping of the rat genome. Specifically, we constructed a framework comparative map of human Chromosome (Chr) 7 and the orthologous regions of the rat genome, as well as two large (>1-Mb) P1-derived artificial chromosome (PAC) based physical maps. Generation of these physical maps involved the use of mouse derived probes that cross-hybridized with rat PAC clones. The first PAC map encompasses the cystic fibrosis transmembrane conductance regulator gene (Cftr), while the second map allows a three-species comparison of a genomic region containing intra- and inter-chromosomal evolutionary rearrangements. The studies reported here further demonstrate that cross-species hybridization between related animals, such as rat and mouse, can be readily used for the targeted construction of clone-based physical maps, thereby accelerating the analysis of biologically interesting regions of vertebrate genomes. PMID- 11420613 TI - Marsupials and Eutherians reunited: genetic evidence for the Theria hypothesis of mammalian evolution. AB - The three living monophyletic divisions of Class Mammalia are the Prototheria (monotremes), Metatheria (marsupials), and Eutheria ('placental' mammals). Determining the sister relationships among these three groups is the most fundamental question in mammalian evolution. Phylogenetic comparison of these mammals by either anatomy or mitochondrial DNA has resulted in two conflicting hypotheses, Theria and Marsupionta, and has fueled a "genes versus morphology" controversy. We have cloned and analyzed a large nuclear gene, the mannose 6 phosphate/insulin-like growth factor II receptor (M6P/IGF2R), from representatives of all three mammalian groups, including platypus, echidna, opossum, wallaby, hedgehog, mouse, rat, rabbit, cow, pig, bat, tree shrew, colugo, ringtail lemur, and human. Statistical analysis of this nuclear gene unambiguously supports the morphology-based Theria hypothesis that excludes monotremes from a clade of marsupials and eutherians. The M6P/IGF2R was also able to resolve the finer structure of the eutherian mammalian family tree. In particular, our analyses support sister group relationships between lagomorphs and rodents, and between the primates and Dermoptera. Statistical support for the grouping of the hedgehog with Feruungulata and Chiroptera was also strong. PMID- 11420614 TI - Assignment of 64 genes expressed in 28-day-old pig embryo to radiation hybrid map. AB - A swine resource family was constructed at the National Institute of Animal Industry, Japan, in order to determine the genetic regions responsible for economically important traits, including fetus development. To identify genes expressed in the early stage of embryo development, we cataloged and mapped genes expressed in a 28-day-old normal pig embryo. In this effort, we have mapped 64 genes, which have map information in human genome onto a swine radiation hybrid (RH) map, IMpRH. These mappings provided additional chromosomal homologies between swine and human to improve the comparative map between the two species. The distribution of the genes assigned to swine chromosomes are as follows: 9 genes were assigned on SSC6; 6 genes each assigned on SSC5 and SSC14; 5 genes each assigned on SSC3, SSC4, and SSC8; 4 genes each assigned on SSC1, SSC7, SSC9, and SSC15; 3 genes each assigned on SSC2, SSC13 and SSCX; and 1 gene each assigned on SSC10, SSC11, and SSC16. Moreover, the present findings revealed 18 new chromosomal homologies between pig and human. Briefly, SSC3 regions were indicated to correspond with HSA1 and HSA10; SSC4 with HSA6; SSC5 with HSA2, HSA15, and HSA16; SSC6 with HSA3, HSA6, and HSA20; SSC7 with HSA11; SSC8 with HSA3, HSA6, and HSA7; SSC9 with HSA8; SSC13 with HSA1; SSC14 with HSA13; SSC15 with HSA19; SSC16 with HSA9. PMID- 11420615 TI - Directed isolation and mapping of microsatellites from swine Chromosome 1q telomeric region through microdissection and RH mapping. AB - Several quantitative trait loci (QTLs) (vertebrate number, birth weight, age at puberty, growth rate, gestation length, and backfat depth) have been independently mapped to the distal region of swine Chromosome (SSC) 1q in several resource populations. In order to improve the map resolution and refine these QTLs more precisely on SSC1q, we have isolated and mapped additional microsatellites (ms), using chromosome microdissection and radiation hybrid (RH) mapping. Five copies of the telomeric region of SSC1q were microdissected from metaphase spreads and pooled. The chromosomal fragment DNA was randomly amplified by using degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR), enriched for ms, and subcloned into a PCR vector. Screening of subsequent clones with ms probes identified 23 unique ms sequences. Fifteen of these (65%) were subjected to radiation hybrid (RH) mapping by using the INRA-University of Minnesota porcine RH panel (IMpRH); and the remaining eight were not suited for the RH mapping. Twelve microsatellites were assigned to SSC1q telomeric region of IMpRH map (LOD >6), and three remain unlinked (LOD <6). Out of the 15 microsatellite markers, 9 were polymorphic in NIAI reference population based on the Meishan and Gottingen miniature pig. In summary, we have used microdissection and radiation hybrid mapping to clone and map 12 new microsatellites to the swine gene map to increase the resolution of SSC1q in the region of known QTLs. PMID- 11420616 TI - Isolation and characterization of hepatic and intestinal expressed sequence tags potentially involved in trait differentiation between cows of different metabolic type. AB - mRNA differential display was applied to identify hepatic and intestinal expressed sequence tags (ESTs) in lactating cows of different metabolic types (milk type, meat/milk type, meat type) that are potentially associated with energy turnover and involved in the regulation of these processes. Altogether, 277 ESTs (liver: 161, intestine: 116) were identified. For 150 transcripts (liver: 99, intestine: 51), the sequences showed similarity to previously described genes and ESTs. Many of these homologous sequences are reported to be involved in hepatic metabolism. Ninety-four ESTs (liver: 43, intestine: 51) did not match with any database entries. Semi-quantitative RT-PCR revealed quantitative differences in transcript represented by randomly chosen ESTs in liver samples of animals of the Holstein and Charolais breeds. One hundred twenty two ESTs were mapped physically by using a bovine-hamster somatic cell hybrid panel (SCP) and a 5000-rad bovine whole genome radiation hybrid panel (WGRH). These ESTs were assigned to the bovine syntenic groups and positioned in the recently established RH-based ordered comparative map of the cattle and human genomes. The mapped, differentially expressed sequence tags are a useful prerequisite for cloning of genetic variation underlying economic traits. PMID- 11420617 TI - Expression and conservation of processed copies of the RBMX gene. AB - RBMX and RBMY are members of an ancient pair of genes located on the sex chromosomes that encode RNA-binding proteins involved in splicing. These genes have differentiated and evolved separately on the X and Y Chromosomes. RBMY has acquired a testis-specific function, whereas, as shown here, RBMX is ubiquitously expressed and is subject to X inactivation. We have also found that multiple processed copies of RBMX are present in the human genome. RBMX-like sequences (RBMXLs) located on human Chrs 1, 4, 6, 9 (9p13 and 9p24), 11, 20, and X lack introns and thus probably result from retroposition events. We found RBMXLs to be conserved in primates and great apes at corresponding chromosomal locations, indicating that they arose prior to the divergence of human. Some of the RBMXLs show insertions, deletions, and stop codons, which would probably result in nonfunctional proteins. The RBMXL on Chr 20 is deleted in some individuals. Two of the largely intact RBMXLs, located on Chrs 1 and 9p13, are expressed in different tissues and may encode novel proteins involved in splicing in a tissue specific manner. The RBMXL located at 9p13 is specifically expressed in testis, and to a lesser extent in brain, and may therefore play a role in testis function. This autosomal, testis-specific copy of RBMX could potentially compensate for RBMX that is presumably inactivated in male germ cells, in a manner analogous to autosomal retroposed copies of other X-linked genes. PMID- 11420618 TI - Quantitative trait loci influencing morphine antinociception in four mapping populations. AB - Analgesia (pain reduction, or antinociception) is a classical and clinically important effect of morphine administration, and in rodent models sensitivity to morphine has been shown to be strongly influenced by genotype. For example, several studies have reported marked differences in morphine antinociception between the insensitive C57BL/6 (B6) and sensitive DBA/2 (D2) inbred mouse strains on the hot-plate assay. This prompted the present genome-wide search for quantitative trait loci (QTLs) that are chromosomal sites influencing the magnitude of antinociception, by using four mapping populations derived from the B6 and D2 progenitor inbred strains. These four were the BXD recombinant inbred (RI) strain set, an F2 (B6D2F2) population, short-term selective breeding for antinociception from a B6D2F2 founding population, and incipient or completed congenic strains. In the BXD RI set and in the B6D2F2, a genome-wide search identified 10-12 provisional QTLs at a nominal p <.05. The other populations were subsequently used as confirmation steps to test each of the provisional QTL regions. Based on all available mapping populations, four QTLs emerged as significant (p <.00005) on proximal Chromosome (Chr) 1 (females only), proximal Chr 9 (females only), mid Chr 9, and proximal Chr 10. The Chr 10 QTL comaps to the same region as the micro-opioid receptor gene (Oprm); this receptor is a known mediator of morphine's antinociceptive effects. The Chr 1 QTL was evident only in females and comapped with the kappa-opioid receptor gene, Oprk. PMID- 11420619 TI - Evolutionary conservation, developmental expression, and genomic mapping of mammalian Twisted gastrulation. AB - The twisted gastrulation gene (tsg) encodes a secreted protein required for the correct specification of dorsal midline cell fate during gastrulation in Drosophila. We report that tsg homologs from human, mouse, zebrafish, and Xenopus share 72-98% identity at the amino acid level and retain all 24 cysteine residues from Drosophila. In contrast to Drosophila where tsg expression is limited to early embryos, expression is found throughout mouse and human development. In Drosophila, tsg acts in synergy with decapentaplegic (dpp), a member of the TGF beta family of secreted proteins. The vertebrate orthologs of dpp, BMP-2 and -4, are crucial for gastrulation and neural induction, and aberrant signaling by BMPs and other TGF-beta family members results in developmental defects including holoprosencephaly (HPE). Interestingly, human TSG maps to the HPE4 locus on Chromosome 18p11.3, and our analysis places the gene within 5 Mbp of TG interacting factor (TGIF). PMID- 11420620 TI - Comparative mapping of BTA15 and HSA11 including a region containing a QTL for meat tenderness. AB - The starting point of the present study was the reported identification of a chromosomal region on bovine Chromosome (Chr) 15 (BTA15) carrying loci affecting meat tenderness. A comparative linkage map of BTA15 and human Chr 11 (HSA11) was constructed to identify potential positional candidate genes and to provide a resource of genetic markers to support marker-assisted selection (MAS). Relative rearrangements between the bovine and human genomes for these chromosomes are the most complex observed in comparative mapping between the two species, with nine alternating blocks of conserved synteny between HSA11 and bovine Chrs 15 and 29. The results of this study were the addition of nine genes to the HSA11/BTA15 comparative linkage map, and development of five microsatellite markers within the quantitative trait locus (QTL) interval. One gene with known effects on muscle development (MYOD1) was mapped to the interval. A second gene (CALCA) involved in regulation of calcium levels, a key factor in postmortem tenderization, also mapped within the interval. Refinement of the comparative map and QTL position will reduce the interval on the human transcription map to be scanned in search of candidates, reducing the effort and resources required to identify the allelic variation responsible for the genetic effect. PMID- 11420621 TI - Comparative analysis of aryl-hydrocarbon receptor interacting protein-like 1 (Aipl1), a gene associated with inherited retinal disease in humans. AB - Mutations in AIPL1 cause Leber congenital amaurosis (LCA), the most severe form of inherited blindness in children; however, the function of this protein in normal vision remains unknown. To determine amino acid subsequences likely to be important for function, we have compared the protein sequence of several species. Sequence conservation is highest across the three Aipl1 tetratricopeptide (TPR) motifs and extends across the protein, except for a proline-rich amino acid sequence present only at the C-terminus of primate Aipl1. The length of the proline-rich region varies within primates; however, the length differences between human and primate Aipl1 do not correlate with evolutionary distance. These observations reinforce the importance of the TPR domains for function, the similarity of Aipl1 to a family of proteins that act as molecular chaperones, and the importance of comparative sequencing data for determination of whether AIPL1 sequence variants in patients are likely to cause retinopathy. PMID- 11420622 TI - Assessment of genetic variation explained by markers for wool traits in sheep via a segment mapping approach. PMID- 11420624 TI - A comparison of diagnostic accuracy of a newly developed photofluorographic panoramic system with conventional panoramic radiography. AB - OBJECTIVES: To compare the diagnostic accuracy of a prototype photofluorographic panoramic system with that of a conventional panoramic system. METHODS: We used 14 human skulls with four artificial apical bone defects in each and 13 clinical cases with 21 periapical lesions. Six dental radiologists and six senior dental students rated the presence of defects on each image on a five-point scale. Receiver Operating Characteristic (ROC) analysis was performed and the area under each curve (Az) was compared as an index of accuracy. Statistical significance was set at P<0.05. RESULTS: There were no significant differences between the photofluorographic and the conventional panoramic systems (P=0.074) in detecting artificial apical defects. However, there were significant differences (P=0.03) in detecting natural defects. CONCLUSIONS: The diagnostic accuracy of the prototype photofluorographic panoramic system appears comparable with that of the conventional panoramic system. However, variability may arise from the observers' diagnostic experience and training. PMID- 11420625 TI - Comparison of computed tomography with conventional radiography for midfacial fractures. AB - OBJECTIVE: To compare the clinical effectiveness of computed tomography (CT) with conventional radiography in midfacial fractures. METHODS: The conventional radiographs (CM) and CT scans of 40 consecutive patients with complex midfacial fractures were assessed independently by two examiners. The number and site of fractures of the orbit, zygoma and maxilla were compared by the Wilcoxon Matched Pairs Signed Rank test. The best method for classification of the fracture was determined. RESULTS: Coronal CT (CCT) proved superior in the diagnosis of orbital fractures (P<0.001). There was no significant difference between any of the imaging methods for fractures of the zygoma. Axial CT (ACT) was the most effective method in imaging of maxillary fractures (ACT-CM; P<0.001, ACT-CCT; P<0.01). CCT was the most useful in classification of orbital and maxillary fracture. CONCLUSION: CCT is superior to CM for the assessment of complex midface fractures PMID- 11420626 TI - Sensitometric comparison of speed group E and F dental radiographic films. AB - OBJECTIVE: To evaluate the sensitometric characteristics of Insight, (Eastman Kodak, Rochester, NY, USA) a new F-speed film, in fresh and depleted processing solutions and compare them with Ektaspeed Plus. METHODS: Two sets each of Insight (IP) and Ektaspeed Plus (EP) films were exposed to radiation levels ranging from 10.7 to 685.2 microGy. One set of films was processed in fresh chemicals while the other set was processed in solutions that had been used for 5 days to process over 500 radiographs. Unexposed films of both types were processed in both solutions to determine base-plus-fog density. Speed and contrast were measured according to ISO definitions and at other levels of density. RESULTS: IP was in speed group F as measured at optical density 1 above base-plus-fog when processed under both conditions. It was 25% faster than EP when both were processed in new solutions and 35% faster in the old solutions, permitting a 20-24% reduction in exposure time. The speeds of both film types decreased when processed in used solutions, but the decrease was smaller for IP than for EP. Speeds at other density levels were greater for IP than EP. Contrast as defined by ISO, and over other density ranges, was similar for both films. CONCLUSIONS: Insight is an F speed film with a speed at least 25% greater than Ektaspeed Plus. IP is more resistant than Ektaspeed Plus to decreases in speed when processed in used chemicals. Contrast of IP and EP is comparable over several density ranges. PMID- 11420627 TI - Volumetry of simulated bone grafts in the edentulous maxilla by computed tomography: an experimental study. AB - OBJECTIVE: To determine the precision of CT for calculating the volume of bone grafts to the maxilla. METHODS: Plaster of Paris was used to simulate bone as onlay and inlay grafts to the maxillary alveolus and sinus in a dry skull. Two mm thick axial CT scans were obtained. The area of the graft on each slice was measured and volumes calculated according to the formula V(graft)=Sigma of areas x the thickness of each section. Scans were repeated with the skull at approximately +5 degrees and -5 degrees to the horizontal plane and the calculations repeated. The real volume was calculated by the displaced water technique. Precision was estimated as reliability coefficient. RESULTS: The reliability coefficients for the right and left inlay and the onlay grafts were 0.97, 0.6 and 0.96 respectively. CONCLUSION: Calculations of the volume of both inlay and onlay grafts to the maxilla from axial CT is precise. PMID- 11420628 TI - A clinical study of changes in the volume of bone grafts in the atrophic maxilla. AB - OBJECTIVES: To evaluate the volumetric changes in bone grafts to the severely atrophic edentulous maxilla over 6 months. METHODS: Ten edentulous patients received autogenous bone as buccal onlay grafts to the maxilla and bilaterally particulate bone grafts to the maxillary sinus. Two mm contiguous axial CT scans were obtained 1 to 2 weeks and 6 to 7 months postoperatively. The area of each graft was measured and the volume calculated according to the formula V(graft)=Sigma of areas x the thickness of each section. RESULTS: The volume of the inlay and onlay grafts was reduced by an average of 49.5 and 47% respectively of the initial volume. The coefficient of variation was greater in the onlay group (0.79) compared with the inlay group (0.38). CONCLUSIONS: The volumetric changes in buccal onlay and sinus inlay grafts after 6 months of healing seem similar although there was a wide range in both groups. PMID- 11420629 TI - A study of the relationship between the position of the condylar head and displacement of the temporomandibular joint disk. AB - OBJECTIVES: To examine the possible relationship between condylar position and disk displacement in the TMJ. METHODS: Forty-eight joints with no disk displacement (NDD), 84 joints with a reducible displaced disk (RDD) and 99 joints with a permanently displaced disk (PDD) were selected. The positions of the condyle and TMJ disk were calculated from lateral MR images. RESULTS: There were significant differences in condylar position between the joints with NDD and RDD, while there was no difference between those with NDD and PDD (Kruskal-Wallis and post-hoc test, P<0.05). There was also a significant correlation between the position of the condylar and the displaced disk (Spearman's rank correlation, P<0.05). The condyle was located more anteriorly with advancing disk displacement. CONCLUSIONS: We suggest that if the disk displacement is slight, the condyle is displaced posteriorly: as the disk displacement becomes more severe (i.e. more anteriorly displaced), the condyle returns to the concentric position. PMID- 11420630 TI - Scanning resolution and the detection of approximal caries. AB - OBJECTIVE: To determine a proper scanning resolution for digitizing bitewing radiographs in the detection of approximal caries. METHODS: Fifty-two premolars and 48 molars were mounted in blocks and imaged on conventional film (Ektaspeed Plus, Eastman-Kodak, Rochester, NY USA) simulating a bitewing projection. The 15 bitewing radiographs were then scanned with a flatbed scanner at three resolutions 150, 300 and 600 d.p.i. The digitized images were displayed in random order on a high-resolution cathode ray tube monitor. Ten observers assessed the caries status of 200 approximal surfaces. They scored lesion presence on a 5 point confidence scale and depth on a 3-point scale. The observer's scores were compared with the results from a histological examination. Data were analysed using analysis of variance, by calculating signed observer error, absolute observer error and observer confidence. RESULTS: Lesion depth had a significant effect on confidence of lesion recognition. The main effect of resolution and the interaction between resolution and lesion depth were significant. Pair-wise comparison showed a significant difference between resolutions in case of sound surfaces and surfaces with dentinal lesions for absolute error. The confidence increased as the resolution increased but no significant difference was found between 300 and 600 d.p.i. The best score for depth estimation was obtained at the 300 d.p.i. scanning resolution. CONCLUSIONS: When bitewing radiographs are scanned with a flatbed scanner, a resolution of 300 d.p.i. seems the best choice. At this resolution the digital file size is manageable without significant loss of the information necessary for caries diagnosis. PMID- 11420631 TI - Integration of a picture archiving and communication system with videocapture and computed radiography in a dental hospital. AB - A method for the integration of a picture archiving and communication system (PACS) at Kyushu University Dental Hospital with radiological information (RIS) and hospital information (HIS) systems is described. CT, US and DSA from different manufacturers were integrated by videocapture and then subsequently integrated with computed radiography (CR) by means of DICOM. The approximate amount of data stored each month on optical discs is 2 GB. The system does not incorporate intra-oral radiography. PMID- 11420632 TI - Analysis of fractal dimensions of rat bones from film and digital images. AB - OBJECTIVES: (1) To compare the effect of two different intra-oral image receptors on estimates of fractal dimension; and (2) to determine the variations in fractal dimensions between the femur, tibia and humerus of the rat and between their proximal, middle and distal regions. METHODS: The left femur, tibia and humerus from 24 4-6-month-old Sprague-Dawley rats were radiographed using intra-oral film and a charge-coupled device (CCD). Films were digitized at a pixel density comparable to the CCD using a flat-bed scanner. Square regions of interest were selected from proximal, middle, and distal regions of each bone. Fractal dimensions were estimated from the slope of regression lines fitted to plots of log power against log spatial frequency. RESULTS: The fractal dimensions estimates from digitized films were significantly greater than those produced from the CCD (P=0.0008). Estimated fractal dimensions of three types of bone were not significantly different (P=0.0544); however, the three regions of bones were significantly different (P=0.0239). The fractal dimensions estimated from radiographs of the proximal and distal regions of the bones were lower than comparable estimates obtained from the middle region. CONCLUSIONS: Different types of image receptors significantly affect estimates of fractal dimension. There was no difference in the fractal dimensions of the different bones but the three regions differed significantly. PMID- 11420633 TI - A rare case of intraosseous polymorphous low-grade adenocarcinoma (PLGA) of the maxilla. AB - A rare case of intraosseous, polymorphous low-grade adenocarcinoma (PLGA) of the maxilla is presented. The lesion appeared to be cystic radiographically and the only finding which suggested malignancy was an irregular cortical border. PMID- 11420634 TI - Ectopic parotid gland as an unusual cause of cheek swelling. AB - There are numerous causes for chronic cheek swelling, including masseteric hypertrophy, lymphadenopathy, diffuse inflammatory changes and neoplasia. We report an unusual case where the swelling was the result of ectopic parotid tissue. PMID- 11420635 TI - A book review dissected. PMID- 11420637 TI - Development of a suspension packaging cell line for production of high titre, serum-resistant murine leukemia virus vectors. AB - To date, only adherent cell lines have been used for the generation of packaging cells for the production of type C retrovirus vectors. The large-scale production of high titre retrovirus vectors could benefit from the development of packaging cells growing in suspension. Here, we describe the ability of two different lymphoid cell lines, one B- and one T-lymphoblastoid cell line (Namalwa and CEM, respectively), to produce MLV-based vectors. Upon transfection with a third generation packaging construct, the virus particle production by Namalwa cells was characterised by low RT-activity, and by CEM cells as high RT activity as previously established adherent packaging cells. An amphotropic packaging cell line (CEMFLYA) was therefore established from CEM cells. Upon introduction of a lacZ vector genome, the novel packaging cell line produced vector particles routinely in the region of 10(7) infectious units/ml. The vectors were helper free and highly stable in fresh human serum. The potential for scaled up vector production was demonstrated by continuous culture of the new packaging cells for 14 days in a 250 ml spinner flask. These suspension packaging cells should be applicable to large bioreactor systems to bulk produce high titre, complement resistant retrovirus vectors for gene therapy. PMID- 11420638 TI - Intravenous infusion of a replication-selective adenovirus (ONYX-015) in cancer patients: safety, feasibility and biological activity. AB - Although genetically engineered adenoviruses hold promise for the treatment of cancer, clinical trial reports have utilized intratumoral injection to date. To determine the feasibility of intravenous delivery of ONYX-015, an E1B-55kD gene deleted replication selective adenovirus with demonstrated clinical safety and antitumoral activity following intratumoral injection, we performed a clinical trial in patients with metastatic solid tumors. ONYX-015 was infused intravenously at escalating doses of 2 x 10(10) to 2 x 10(13) particles via weekly infusion within 21-day cycles in 10 patients with advanced carcinoma metastatic to the lung. No dose-limiting toxicity was identified. Mild to moderate fever, rigors and a dose-dependent transient transaminitis were the most common adverse events. Neutralizing antibody titers significantly increased within 3 weeks in all patients. IL-6, gamma-IFN, TNF-alpha and IL-10 increased within 24 h following treatment. Evidence of viral replication was detectable in three of four patients receiving ONYX-015 at doses > or = 2 x 10(12) particles and intratumoral replication was confirmed in one patient. In conclusion, intravenous infusion of ONYX-015 was well tolerated at doses up to 2 x 10(13) particles and infection of metastatic pulmonary sites with subsequent intratumoral viral replication was seen. The intravenous administration of genetically altered adenovirus is a feasible approach. PMID- 11420639 TI - Transfection of urothelial cells using methyl-beta-cyclodextrin solubilized cholesterol and Dotap. AB - the murine urothelial cell line, mb49 was transfected with the reporter gene pcmvlacz using a number of commercial transfection agents. the transfection efficiency of these agents, as determined by beta-galactosidase activity, is in the order of dotap>superfect>Fugene. The addition of methyl-beta-cyclodextrin solubilized cholesterol (MBC) to Dotap and Superfect further improved their transfection efficiency by 3.8-fold and 2.6-fold, respectively. beta Galactosidase activity was detectable within 1 h of transfection and peaked at 48 h. Nuclear and cytoplasmic separation showed that with Dotap + methyl-beta cyclodextrin solubilized cholesterol (DMBC), the DNA plasmid complex was found in both the nucleus and the cytoplasm. In vivo, murine bladders were transfected with an intravesical instillation of DMBC + DNA for 2 h. Two days later the bladder, lungs, liver, spleen and heart were assayed for the presence of the beta galactosidase gene by staining and PCR. Expression of the gene was confined to the bladder. Both in vitro and in vivo expression was observed after as little as a 15 min exposure to DMBC:DNA. Expression of the marker gene was present up to 30 days after transfection in vivo. From our data it appears that DMBC is the best nonviral agent for the transfection of urothelial cells in vitro and in vivo. PMID- 11420640 TI - Expression of interleukin-4 but not of interleukin-10 from a replicative herpes simplex virus type 1 viral vector precludes experimental allergic encephalomyelitis. AB - We have used interleukin (IL)-4 and -10-producing HSV-1 gamma(1)34.5 deletion viruses in gene therapy of a BALB/c model of experimental allergic encephalomyelitis (EAE), a T cell-mediated demyelinating disease of the central nervous system. It is known that in EAE of mice the Th2-type cytokines are down regulated and the Th1-type cytokines up-regulated during the onset and relapse of the disease. Therefore, we tested two HSV-1 recombinants expressing the Th2-type cytokines IL-4 and IL-10. The recombinant viruses were injected intracranially (i.c.) in BALB/c mice 6 days after induction of EAE. As control groups we used mice without any infection, mice infected with backbone virus R3659 and mock infected mice. Weights and symptoms of the mice were recorded daily and the tissue specimens were collected at specific time-points. The results indicate that the intracranial infection with IL-4-producing virus (1) precludes EAE symptoms, (2) protects the spinal cord from massive leukocyte infiltrations and (3) prevents demyelination and axonal loss. The IL-10-expressing virus R8308 did not have a similar favorable effect on the recovery of the mice as did the IL-4 virus R8306. PMID- 11420641 TI - Transplanted primary neonatal myoblasts can give rise to functional satellite cells as identified using the Myf5nlacZl+ mouse. AB - Myoblast transplantation is a potential therapeutic approach for the genetic modification of host skeletal muscle tissue. To be considered an effective, long lived method of delivery, however, it is essential that at least a proportion of the transplanted cells also retain their proliferative potential. We sought to investigate whether transplanted neonatal myoblasts can contribute to the satellite cell compartment of adult skeletal muscle by using the Myf5nlacZ/+ mouse. The Myf5nlacZ/+ mouse has nlacZ targeted to the Myf5 locus resulting in beta-galactosidase activity in quiescent satellite cells. Following transplantation, beta-galactosidase-labelled nuclei were detected in host muscles, showing that donor cells had been incorporated. Significantly, beta galactosidase-positive, and therefore donor-derived, satellite cells were detected. When placed in culture, beta-galactosidase marked myogenic cells emanated from the parent fibre. These observations demonstrate that cell transplantation not only results in the incorporation of donor nuclei into the host muscle syncytia, but also that the donor cells can become functional satellite cells. The Myf5nlacZ/+ mouse therefore provides a novel and specific marker for determining the contribution of transplanted cells to the satellite cell pool. PMID- 11420642 TI - Angiographically guided utero-placental gene transfer in rabbits with adenoviruses, plasmid/liposomes and plasmid/polyethyleneimine complexes. AB - We examined the feasibility of gene transfer to rabbit placenta using adenoviruses, plasmid/liposomes and plasmid/polyethyleneimine (PEI) complexes. Pregnant New Zealand White rabbits (n = 17) were anesthetized and local gene transfer was done via a catheter inserted in uterine arteries under direct angiographic control. Either nuclear targeted LacZ adenoviruses (1.0 x 10(10) p.f.u.), nuclear targeted LacZ plasmid (500 microg)/liposome (DOTMA:DOPE 1:1) complexes or nuclear targeted LacZ plasmid (250 microg)/PEI (25 kDa) complexes (charge ratio +/-4) were used. Animals were killed 3 days later and detection of the transgene expression was done by X-gal staining and RT-PCR. Adenovirus mediated gene transfer resulted in a high transfection efficiency (34 +/- 10%) in placental trophoplastic cells. Very little, if any, transfection was seen in fetal membranes. Plasmid/liposomes and plasmid/PEI complexes led to a very low (<0.01%) transfection efficiency in trophoblastic cells, but some transfection was seen in fetal membranes. A total of 25 fetuses were analyzed for the presence of transgene at the time of death. In most fetuses expression of the LacZ gene was below the sensitivity of the X-gal staining, but expression was detected by PCR in 50%, 50% and 42% of the analyzed fetuses after adenoviral, plasmid/PEI and plasmid/liposome gene transfer, respectively. No major inflammatory changes were present in the transfected placentas as analyzed by general histology and macrophage- and T cell-specific immunostainings. We conclude that catheter mediated intravascular gene transfer with adenoviruses can be used for the transfection of placental trophoplastic cells, but plasmid complexes are inefficient for this purpose. However, selective angiographically guided gene transfer also led to leakage of the vector to fetuses. Therefore, if gene therapy is developed for the treatment of placental disorders, the gene-vector combination should not be harmful to the fetus and the expression of the transgene should only occur in placenta. PMID- 11420643 TI - A modified adenovirus can transfect cochlear hair cells in vivo without compromising cochlear function. AB - The loss of cochlear hair cells, or the loss of their capacity to transduce acoustic signals, is believed to be the underlying mechanism in many forms of hearing loss. To develop viral vectors that allow for the introduction of genes directly into the cochleae of adult animals, replication-deficient (E1(-), E3(-)) and replication-defective (E1(-), E3(-), pol(-)) adenovirus vectors were used to transduce the bacterial beta-galactosidase gene into the hair cells of the guinea pig cochlea in vivo. Distortion product otoacoustic emissions, which monitor the functional status of outer hair cells, were measured throughout the viral infection periods to identify hair cell ototoxicity. The results demonstrated that the use of the (E1(-), E3(-)) adenovirus vectors containing CMV-driven LacZ, compromised cochlear function when gradually introduced into scala tympani via an osmotic pump. However, when (E1(-), E3(-), pol(-)) adenoviral vectors containing CMV-driven LacZ were used to transduce cochlear hair cells, there was no loss of cochlear function over the frequency regions tested, and beta-galactosidase (beta gal) was detected in over 80% of all hair cells. Development of a viral vector that infects cochlear hair cells without virus-induced ototoxic effects is crucial for gene replacement strategies to treat certain forms of inherited deafness and for otoprotective strategies to prevent hair cell losses to treat progressive hearing disorders. Moreover, in vivo (E1(-), E3(-), pol(-)) adenovirus mediated gene-transfer techniques applied to adult guinea pig cochleae may be useful in testing several hypotheses concerning what roles specific genes play in normal cochlear function. PMID- 11420644 TI - 3PO, a novel nonviral gene delivery system using engineered Ad5 penton proteins. AB - This study describes the development of 3PO, a nonviral, protein-based gene delivery vector which utilizes the highly evolved cell-binding, cell-entry and intracellular transport functions of the adenovirus serotype 5 (Ad5) capsid penton protein. A penton fusion protein containing a polylysine sequence was produced by recombinant methods and tested for gene delivery capability. As the protein itself is known to bind integrins through a conserved consensus motif, the penton inherently possesses the ability to bind and enter cells through receptor-mediated internalization. The ability to lyse the cellular endosome encapsulating internalized receptors is also attributed to the penton. The recombinant protein gains the additional function of DNA binding and transport with the appendage of a polylysine motif. This protein retains the ability to form pentamers and mediates delivery of a reporter gene to cultured cells. Interference by oligopeptides bearing the integrin binding motif suggests that delivery is mediated specifically through integrin receptor binding and internalization. The addition of protamine to penton-DNA complexes allows gene delivery in the presence of serum. PMID- 11420645 TI - Protection against ischemia/reperfusion injury and myocardial dysfunction by antisense-oligodeoxynucleotide directed at angiotensin-converting enzyme mRNA. AB - Angiotensin-converting enzyme (ACE) activity in the myocardium and angiotensin-II (Ang-II) levels in plasma increase after myocardial ischemia, which lead to exacerbation of myocardial injury and cardiac dysfunction. We examined the protective role of novel antisense-oligodeoxynucleotide (AS-ODN) directed at ACE mRNA in myocardial ischemic injury. Sprague-Dawley rats were treated with ACE-AS ODN (200 microg per rat, n = 8, i.v.) or inverted-ODN (IN-ODN, 200 microg per rat, n = 8, i.v.), given with 600 microg per rat of liposome DOTAP/DOPE. Hearts from AS-ODN- or IN-ODN-treated rats were excised, perfused in vitro, and subjected to 25 min of global ischemia followed by 30 min of reperfusion. Parallel groups of rats were given ACE inhibitor captopril (5 mg/kg, n = 8) or saline (n = 8) before excising the hearts. Ischemia/reperfusion resulted in myocardial dysfunction (increase in coronary perfusion pressure and LV end diastolic pressure and a decrease in developed LV pressure) in the saline-treated rats. Myocardial dysfunction was associated with evidence of lipid peroxidation and enzyme leakage (MDA and LDH levels in the myocardium) and up-regulation of ACE protein expression. Administration of AS-ODN or captopril, but not IN-ODN, reduced Ang-II levels in the plasma, decreased ischemia/reperfusion-mediated cardiac functional deterioration and lipid peroxidation, and preserved LDH in the myocardium (all P < 0.05 versus the saline group). AS-ODN and captopril had equipotent effects on cardiac dynamics. ACE protein expression (western blot) was decreased in the hearts of the AS-ODN-treated group, but not in IN-ODN-treated rat hearts. In contrast, ACE protein expression was significantly increased in captopril-treated rat hearts. These observations suggest that AS-ODN directed at ACE mRNA can ameliorate myocardial dysfunction and injury after ischemia/reperfusion, and its use is associated with decreased expression of ACE protein in the ischemic myocardium. PMID- 11420646 TI - Retroviral vector-mediated expression of HoxB4 in hematopoietic cells using a novel coexpression strategy. AB - Retroviral vector-mediated expression of the homeoboxgene, HoxB4, in hematopoietic cells has been reported to mediate a benign expansion of gene modified hematopoietic stem and precursor cells in vivo. In the present study, we used a novel coexpression strategy for coordinated expression of HoxB4 along with a cytoplasmic protein from a retroviral vector. The novel coexpression strategy, based on cotranslational protein separation mediated by the 2A sequence of foot and-mouth disease virus (FMDV), allows an indirect quantification of HoxB4 expression levels when inserting a reporter such as the enhanced green fluorescent protein (GFP) in the retroviral vector. Presence of the 2A sequence did not interfere with the correct subcellular localization of HoxB4 (nuclear) and GFP (cytoplasmic), nor with the titer of bicistronic vectors, and mediated functional long-term coexpression (at least 1 year) of GFP and HoxB4 after transplantation of transduced mouse bone marrow cells in mice. PMID- 11420647 TI - Phenotypic and genotypic characterization of Cryptosporidium species and isolates. AB - Recent outbreaks of cryptosporidiosis from contaminated water supplies have led to a need for the detection of Cryptosporidium oocysts from various hosts and contaminating sources. The presence of nonpathogenic species or strains of Cryptosporidium is important for diagnostic purposes as there is a potential for false- positive detection of pathogenic parasites. The present review focuses on phenotypic differences and recent advances in genotypic analyses of the genus Cryptosporidium with an emphasis on detecting various isolates and identifying differences in Cryptosporidium parvum and other species in this genus. The information currently available demonstrates important patterns in DNA sequences of Cryptosporidium, and our understanding of macro- and microevolutionary patterns has increased in recent years. However, current knowledge of Cryptosporidium genetic diversity is far from complete, and the large amount of both phenotypic and genotypic data has led to problems in our understanding of the systematics of this genus. PMID- 11420648 TI - Inhibition of bacilli in industrial starches by nisin. AB - The properties of Bacillus coagulans and of other bacilli that contaminate paper and paperboard manufacturing processes were investigated under simulated industrial conditions. Nisin (0.05 to 0.125 microg ml(-1) blocked growth of indigenous bacilli that contaminate sizing starches. B. coagulans starch isolates, B. licheniformis, B. amyloliquefaciens, and B. stearothermophilus grew at > or = 50 degrees C in industrial starch and produced alpha-glucosidase and cyclodextrins. The industrial isolates and reference strains of B. amyloliquefaciens, B. cereus, B. coagulans, B. flexus, B. licheniformis, B. pumilus, B. sporothermodurans, B. stearothermophilus and Alicyclobacillus acidoterrestris were inhibited by < or = 0.125 microg of nisin on agar. B. coagulans and B. stearothermophilus were similarly inhibited by < or = 0.025 microg of nisin ml(-1) and by 3 microg of the biocide DBNPA ml(-1) in industrial starch. B. licheniformis and B. amyloliquefaciens strains were less sensitive. About 40% of nisin added to starch was retained after cooking. Fifty percent of the nisin remained active after 11 h of storage at 60 degrees C. The results show that nisin has potential as a preservative for modified industrial starches. PMID- 11420649 TI - Efficient production of menaquinone (vitamin K2) by a menadione-resistant mutant of Bacillus subtilis. AB - Efficient production of menaquinone (MK) by Bacillus subtilis was achieved. An edible strain of B. subtilis, isolated from the traditional Japanese food natto, was mutated to improve MK productivity. A menadione-resistant mutant producing 30% more MK than its parent strain was obtained. Soybean extract and glycerol were the best nitrogen and carbon sources, respectively, among the sources tested. Addition of yeast extract also increased MK productivity. The maximum concentration of MK reached about 35.0 mg/l after 4 days of culture in a jar fermenter. The pH of the medium decreased to 5.5 after the start of cultivation, then spontaneously increased to 7.7-8.0. This pH change might be important in the production of MK because only small amounts of MK were obtained when pH was controlled at 5.7, 6.0, 7.0, 7.5 or 8.0. PMID- 11420650 TI - Kinetic analysis on formation of poly(3-hydroxybutyrate) from acetic acid by Ralstonia eutropha under chemically defined conditions. AB - Batch cultures of Ralstonia eutropha in chemically defined media with acetic acid (HAc) as the sole carbon source were conducted to investigate acetate utilization, formation of poly(3-hydroxybutyrate) (PHB) and growth of active biomass (ABM) under different carbon to nitrogen (C/N) weight ratios. The specific acetate utilization rate based on ABM approached 0.16 g/g ABM h(-1), which was not affected very much by the extracellular HAc concentration from 1 to 5 g/l, but was affected by the C/N weight ratio. A low C/N ratio or high nitrogen supply sped up the specific acetate utilization rate to produce more ABM and less PHB. A high HAc concentration (>6 g/l), however, depressed acetate utilization as well as the ABM growth and PHB formation. A high cell mass concentration enhanced the tolerance of R. eutropha to the toxicity of HAc at pH 7 to 8.5. The viscosity average molecular size of PHB generally increased first and then declined in batch cultures. Larger PHB molecules and less PHB per ABM were produced at a low C/N ratio with enough nutrient nitrogen than those under a high C/N ratio with less nutrient nitrogen available. PMID- 11420651 TI - Themophilic biofiltration of methanol and alpha-pinene. AB - Biofiltration systems utilizing thermophilic (55 degrees C) bacteria were constructed and tested for the removal of methanol and alpha-pinene--two important volatile organic compounds (VOCs) in the forest products industry. Thermophilic bacterial mixtures that can degrade both methanol and alpha-pinene were obtained via enrichment techniques. Two bench-scale thermophilic biofiltration systems (1085 and 1824 cm3) were used to examine compound removals at different residence times, with influent concentrations of 110 ppmv methanol and 15 ppmv alpha-pinene. At a residence time of 10.85 min, the smaller system had removal efficiencies of >98% for methanol, but only 23% for alpha-pinene. The larger system was operated with the same parameters to evaluate residence time and surfactant effects on compound removals. At a residence time of 18.24 min, both methanol and alpha-pinene removal rates were > or = 95%. However, a-pinene removal dropped to 26% at a residence time of 6.08 min; methanol removal was not affected. Subsequent addition of a surfactant mixture increased a-pinene removal to 94% at the shortest residence time. No residual alpha-pinene was detected with the support medium Celite R-635, indicating that the surfactant may increase mass transfer of alpha-pinene. PMID- 11420652 TI - Biogenic production of cyanide and its application to gold recovery. AB - Chromobacterium violaceum is a cyanogenic (cyanide-producing) microorganism. Cyanide is used on an industrial scale to complex and recover gold from ores or concentrates of ores bearing the precious metal. A potentially useful approach in gold mining operations could be to produce cyanide biologically in relatively small quantities at the ore surface. In this study, C. violaceum grown in nutrient broth formed a biofilm and could complex and solubilize 100% of the gold on glass test slides within 4-7 days. Approximately 50% of the cyanide- recoverable gold could be mobilized from a biooxidized sulfidic-ore concentrate. Complexation of cyanide in solution by gold appeared to have a beneficial effect on cell growth--viable cell counts were nearly two orders of magnitude greater in the presence of gold-coated slides or biooxidized ore substrates than in their absence. C. violaceum was cyanogenic when grown in alternative feedstocks. When grown in a mineral salt solution supplemented with 13.3% v/v swine fecal material (SFM), cells exhibited pigmentation and suspended cell concentrations comparable to cultures grown in nutrient broth. Glycine supplements stimulated production of cyanide in 13.3% v/v SFM. In contrast, glycine was inhibitory when added at the time of inoculation in the more concentrated SFM, decreasing cell numbers and reducing ultimate bulk-solution cyanide concentrations. However, aeration and addition of glycine to stationary phase cells grown on 13.3% v/v SFM anaerobically resulted in rapid production and high concentrations (up to 38 mg l(-1)) of cyanide. This indicates that biogenesis of cyanide may be supported in remote areas using locally produced and inexpensive agricultural feedstocks in place of commercial media. PMID- 11420653 TI - Metabolism of the veterinary fluoroquinolone sarafloxacin by the fungus Mucor ramannianus. AB - To investigate the microbial biotransformation of veterinary fluoroquinolones, Mucor ramannianus was grown in sucrose/peptone broth with sarafloxacin for 18 days. Cultures were extracted with ethyl acetate and extracts were analyzed by liquid chromatography. The two metabolites (26% and 15% of the A280, respectively) were identified by mass and 1H nuclear magnetic resonance spectra as N-acetylsarafloxacin and desethylene-N-acetylsarafloxacin. The biological formation of desethylene-N-acetylsarafloxacin has not been previously observed. PMID- 11420654 TI - Ethanol production from hardwood spent sulfite liquor using an adapted strain of Pichia stipitis. AB - Conditions have been optimized for fermentation of pretreated hardwood spent sulfite liquor (HSSL) using an adapted strain of Pichia stipitis. The pretreatments, consisting of boiling and overliming with Ca(OH)2 of HSSL, to partially remove inhibitors, and adaptation of the yeast strain to HSSL, were both critical for a successful fermentation. Ethanol concentration was increased from 6.7 to 20.2 g l(-1) using adapted P. stipitis (A) and pretreated HSSL. The maximum ethanol yield (Yp/s) and productivity (Qp) were 0.41 g g(-1) and 0.44 g l(-1) h(-1), respectively, at an oxygen transfer rate of 2.0 mmol O2 l(-1) h(-1). The optimized results with this strain were compared to those of other xylose fermenting yeasts and Saccharomyces cerevisiae (SSL-acclimatized) currently used at an industrial plant for the fermentation of spent sulfite liquor. PMID- 11420655 TI - Characterizing the production of a wild-type and benomyl-resistant Fusarium lateritium for biocontrol of Eutypa lata on grapevine. AB - Benomyl-resistant (BR) and wild-type (WT) strains of Fusarium lateritium were examined for their tolerance to benomyl on potato dextrose agar (PDA) containing benomyl and control of the Eutypa lata in grapevine bioassays. The WT strain grew on PDA containing 1 microg/ml benomyl at 13, 26 and 29 degrees C. The BR strain grew on PDA containing 10 microg/ml benomyl at 4 degrees C, on PDA containing 100 microg/ml benomyl at 29 degrees C, and on PDA containing 1,000 microg/ml benomyl at 13 degrees C and 26 degrees C. The BR strain was also able to colonize grapevine segments and control E. lata in the presence of 1,000 microg/ml benomyl. Both strains were amenable to production via liquid fermentation and both achieved 100% control of E. lata in grapevine bioassays. Neither the duration of fermentation nor incubation temperature during grapevine bioassays influenced the efficacy of either strain against E. lata. The results suggest that application of BR F. lateritium alone or in combination with benomyl may provide good control of E. lata. PMID- 11420656 TI - Purification and properties of a thermostable extracellular beta-D-xylosidase produced by a thermotolerant Aspergillus phoenicis. AB - A beta-D-xylosidase was purified from cultures of a thermotolerant strain of Aspergillus phoenicis grown on xylan at 45 degrees C. The enzyme was purified to homogeneity by chromatography on DEAE-cellulose and Sephadex G-100. The purified enzyme was a monomer of molecular mass 132 kDa by gel filtration and SDS-PAGE. Treatment with endoglycosidase H resulted in a protein with a molecular mass of 104 kDa. The enzyme was a glycoprotein with 43.5% carbohydrate content and exhibited a pl of 3.7. Optima of temperature and pH were 75 degrees C and 4.0 4.5, respectively. The activity was stable at 60 degrees C and had a Km of 2.36 mM for p-nitrophenyl-beta-D-xylopiranoside. The enzyme did not exhibit xylanase, cellulase, galactosidase or arabinosidase activities. The purified enzyme was active against natural substrates, such as xylobiose and xylotriose. PMID- 11420657 TI - Predicting vegetative inoculum performance to maximize phytase production in solid-state fermentation using response surface methodology. AB - Microbial phytase is used to reduce the environmental loading of phosphorus from animal production facilities. The limiting factors in the use of this enzyme in animal feeds can be overcome by solid-state fermentation (SSF), which is a promising technology for commercial enzyme production with lower production costs. Inoculum quality and the influence of inoculum quality on phytase production are important factors which need in-depth investigation before scaling up of high-yielding fermentation process. A full factorial experimental design for 240 h with sampling at every 24 h was used to determine the effects of the treatments, inoculum age (plate and liquid culture), media composition and the duration of SSF on the production of fungal biomass and phytase in SSF systems using Aspergillus niger. The optimal treatment combination for maximal phytase production was determined by statistically comparing all treatments at each sampling time. Both 7- and 14-day plate cultures and M1 + medium composition with 72-h-old liquid inoculum treatments resulted in optimal phytase production at 144 h of SSF, which was the shortest duration observed for maximal phytase production. This resulted in maximal phytase production with a mean of 884 +/- 121 U/g substrate, while the maximal phytase production observed at 216 h of SSF (mean phytase activity of 1,008 +/- 121 U/g substrate), with the same treatment combinations, was not statistically significant from that at 144 h of SSF. Phytase production was strongly growth-associated with younger inocula. The significant treatment variables, age of liquid inoculum and the duration of SSF, were used to predict the system response for phytase production using response surface methodology. From the response surface model, the optimal response of the experiment was predicted and the reliability of the prediction was checked with the verification experiment. PMID- 11420658 TI - Effects of acetic acid and lactic acid on the growth of Saccharomyces cerevisiae in a minimal medium. AB - Specific growth rates (mu) of two strains of Saccharomyces cerevisiae decreased exponentially (R2 > 0.9) as the concentrations of acetic acid or lactic acid were increased in minimal media at 30 degrees C. Moreover, the length of the lag phase of each growth curve (h) increased exponentially as increasing concentrations of acetic or lactic acid were added to the media. The minimum inhibitory concentration (MIC) of acetic acid for yeast growth was 0.6% w/v (100 mM) and that of lactic acid was 2.5% w/v (278 mM) for both strains of yeast. However, acetic acid at concentrations as low as 0.05-0.1% w/v and lactic acid at concentrations of 0.2-0.8% w/v begin to stress the yeasts as seen by reduced growth rates and decreased rates of glucose consumption and ethanol production as the concentration of acetic or lactic acid in the media was raised. In the presence of increasing acetic acid, all the glucose in the medium was eventually consumed even though the rates of consumption differed. However, this was not observed in the presence of increasing lactic acid where glucose consumption was extremely protracted even at a concentration of 0.6% w/v (66 mM). A response surface central composite design was used to evaluate the interaction between acetic and lactic acids on the specific growth rate of both yeast strains at 30 degrees C. The data were analysed using the General Linear Models (GLM) procedure. From the analysis, the interaction between acetic acid and lactic acid was statistically significant (P < or = 0.001), i.e., the inhibitory effect of the two acids present together in a medium is highly synergistic. PMID- 11420659 TI - Binary culture biofilm formation by Stenotrophomonas maltophilia and Fusarium oxysporum. AB - Binary culture biofilm formation by Stenotrophomonas maltophilia and Fusarium oxysporum was investigated using the recirculating modified Robbins device batch culture system. Sequential attachment studies were carried out in the Robbins device on PVC and glass surfaces, with each species as either the first or the second colonizer. Different surfaces had no significant effect on total numbers of S. maltophilia and F. oxysporum in the binary population biofilm. The attachment of the second colonizer was not influenced significantly by the previous attachment of the first colonizer. These results were confirmed using scanning electron micrographs. PMID- 11420660 TI - Constitutive activation of Stat3 by the Src and JAK tyrosine kinases participates in growth regulation of human breast carcinoma cells. AB - Constitutive activation of signal transducer and activator of transcription (STAT) proteins has been detected in a wide variety of human primary tumor specimens and tumor cell lines including blood malignancies, head and neck cancer, and breast cancer. We have previously demonstrated a high frequency of Stat3 DNA-binding activity that is constitutively-induced by an unknown mechanism in human breast cancer cell lines possessing elevated EGF receptor (EGF-R) and c Src kinase activities. Using tyrosine kinase selective inhibitors, we show here that Src and JAK family tyrosine kinases cooperate to mediate constitutive Stat3 activation in the absence of EGF stimulation in model human breast cancer cell lines. Inhibition of Src or JAKs results in dose-dependent suppression of Stat3 DNA-binding activity, which is accompanied by growth inhibition and induction of programmed cell death. In addition, transfection of a dominant-negative form of Stat3 leads to growth inhibition involving apoptosis of breast cancer cells. These results indicate that the biological effects of the Src and JAK tyrosine kinase inhibitors are at least partially mediated by blocking Stat3 signaling. While EGF-R kinase activity is not required for constitutive Stat3 activation in breast cancer cells, EGF stimulation further increases STAT DNA-binding activity, consistent with an important role for EGF-R in STAT signaling and malignant progression. Analysis of primary breast tumor specimens from patients with advanced disease revealed that the majority exhibit elevated STAT DNA-binding activity compared to adjacent non-tumor tissues. Our findings, taken together, suggest that tyrosine kinases transduce signals through Stat3 protein that contribute to the growth and survival of human breast cancer cells in culture and potentially in vivo. PMID- 11420661 TI - Phosphoinositide-3 kinase-PKB/Akt pathway activation is involved in fibroblast Rat-1 transformation by human T-cell leukemia virus type I tax. AB - Activated phosphoinositide 3-kinase (PI3K) and its downstream target Akt are essential for the fibroblast transformation induced by many viral products. Tax, encoded by human T-cell leukemia virus type I (HTLV-I), has been demonstrated to induce the transformation of rat fibroblast Rat-1 cell through NF-kappaB activation. By stable transfection of Rat-1 cells with expressing constructs of Tax and its mutant M47, which is defective in HTLV-I LTR transactivation, we selected their transformed clones, which have characteristics of NF-kappaB activation and colony formation beyond the cell monolayer (a malignant phenotype). However, these two characteristics in the transformed clones of Tax and M47 disappear after these cells have been treated with wortmannin, a specific inhibitor of PI3K. Further, increased activity of the PI3K/Akt is observed in the transformed clones of Tax and M47 as compared to the clones of empty vector Neo and the M148, which is defective in NF-kappaB activation and cell transformation. Increased activity of PI5K is present in the transformed clones of both Tax and M47 and in the M148 clone as compared to that in the Neo cell. It is known that the efficiency of Tax-induced cell transformation is not high; a minority of Tax expressing clones show transformation, although the majority of Tax-expressing clones show activated NF-kappaB. A Tax-expressing, nontransformed clone after transfection with an active form of the catalytic subunit of PI3K, p110alpha, becomes transformed. Consistent with these results, a Tax highly-expressing human T-cell line MT2 exhibits both higher polyphosphoinositide turnover and higher activities of PI3K and PI5K than those of Jurkat or MT1 and HTLV-I-negative and a Tax-unexpressing cell line, respectively. These results demonstrate that the activation of the PI3K/Akt signaling pathway, excepting for the NF-kappaB, is also required for the cell transformation induced by Tax. PMID- 11420662 TI - Regulation of p53 expression by the RAS-MAP kinase pathway. AB - Activation of MAP kinase leads to the activation of p53-dependent pathways, and vice-versa. Although the amount of p53 protein increases in response to MAP kinase-dependent signaling, the basis of this increase is not yet fully understood. We have isolated the mutant cell line AP14, defective in p53 expression, from human HT1080 fibrosarcoma cells, which have an activated ras allele. The expression of p53 mRNA and protein is approximately 10-fold lower in AP14 cells than in the parental cells. The high constitutive phosphorylation and activities of the MAP kinases ERK1 and ERK2 in HT1080 cells are greatly reduced in AP14 cells, although the levels of these proteins are unchanged, suggesting that the defect in the mutant cells affects the steady-state phosphorylation of ERK1 and ERK2. Overexpression of ERK2 in AP14 cells restored both MAP kinase activity and p53 expression, and incubation of the mutant cells with the phosphatase inhibitor orthovanadate resulted in strong coordinate elevation of MAP kinase activity and p53 expression. The levels of expression of the p53 regulated gene p21 parallel those of p53 throughout, showing that basal p21 expression depends on p53. The levels of p53 mRNA increased by 5-8-fold when activated ras was introduced into wild-type cells, and the levels of the p53 and p21 proteins decreased substantially in wild-type cells treated with the MEK inhibitor U0216. We conclude that MAP kinase-dependent pathways help to regulate p53 levels by regulating the expression of p53 mRNA. PMID- 11420663 TI - Transcriptional upregulation and activation of p55Cdc via p34(cdc2) in Taxol induced apoptosis. AB - Paclitaxel (Taxol) is a potent and highly effective antineoplastic agent for the treatment of advanced, drug-refractory, metastatic breast cancers. Taxol not only induces tubulin polymerization, stabilizes microtubules, blocks cell cycle progression, and induces apoptosis, but it also alters gene expression. Here, we have identified that Taxol can upregulate expression of the gene encoding the cell cycle protein p55Cdc by using cDNA array technique. Taxol induced p55Cdc mRNA expression through activation of the p55Cdc promoter, which led to increase p55Cdc protein expression. Taxol also activated p55Cdc-associated kinase. In addition, overexpression of the p55Cdc gene resulted in cell death in both HeLa cells and NIH3T3 cells in a dose-dependent manner. A dominant-negative mutant of p34(cdc2) blocked Taxol-induced p55Cdc activation and inhibited p55Cdc-induced and Taxol-induced cell death. Our data suggest that transcriptional upregulation of p55Cdc and activation of p55Cdc by Taxol-mediated p34(cdc2) activation play a critical role in Taxol-induced cell death. PMID- 11420664 TI - A targeted mouse Brca1 mutation removing the last BRCT repeat results in apoptosis and embryonic lethality at the headfold stage. AB - A mouse model with a targeted mutation in the 3' end of the endogenous Brca1 gene, Brca1(1700T), was generated to compare the phenotypic consequences of truncated Brca1 proteins with other mutant Brca1 models reported in the literature to date. Mice heterozygous for the Brca1(1700T) mutation do not show any predisposition to tumorigenesis. Treatment of these mice with ionizing radiation or breeding with Apc, Msh-2 or Tp53 mutant mouse models did not show any change in the tumor phenotype. Like other Brca1 mouse models, the Brca1(1700T) mutation is embryonic lethal in homozygous state. However, homozygous Brca1(1700T) embryos reach the headfold stage but are delayed in their development and fail to turn. Thus, in contrast to Brca1(null) models, the mutant embryos do not undergo growth arrest leading to a developmental block at 6.5 dpc, but continue to proliferate and differentiate until 9.5 dpc. Homozygous embryos die between 9.5-10.5 dpc due to massive apoptosis throughout the embryo. These results indicate that a C-terminal truncating Brca1 mutation removing the last BRCT repeat has a different effect on normal cell function than does the complete absence of Brca1. PMID- 11420665 TI - Diverged nuclear localization of Werner helicase in human and mouse cells. AB - Werner syndrome (WS) is a rare autosomal recessive genetic disorder causing premature aging and rare cancers. A gene responsible for WS (WRN) encodes a protein with 1432 amino acids (a.a.) homologous to the E. coli RecQ-type DNA helicase. Transcriptional activation facilitated nucleolar localization of human WRN protein (hWRNp) and serum starvation induced translocation of hWRNp from the nucleoli to the nucleoplasm in human cultured cells, suggesting a nucleolar nucleoplasm trafficking of hWRNp depending on transcriptional state. Mutant hWRNp lacking the C-terminal 30 a.a. residues (Delta1403-1432) failed to localize in the nucleolus, whereas Delta1405-1432 can migrate into the nucleolus. Here we identify a region putative for nucleolar localization signal (NoLS) containing a sequence of two positively charged amino acids (Arg(1403)-Lys(1404)) in the C terminal area of hWRNp. By contrast, the mouse homolog (mWRNp) exists only in the nucleoplasm. We show that the inability of mWRNp to migrate into the nucleolus is due to a difference of a sequence in the region corresponding to the NoLS of hWRNp. In addition, mouse cells cannot recognize the NoLS of hWRNp. Our study suggests that defect in nucleolar function of hWRNp may be linked to the premature aging which is not observed in mWRN(-/-) mice. PMID- 11420666 TI - Retinoic acid activates p53 in human embryonal carcinoma through retinoid receptor-dependent stimulation of p53 transactivation function. AB - Although retinoids are known to regulate gene transcription by activating retinoid receptors, the targets of retinoid receptors are largely unknown. This study indicates effective all-trans retinoic acid (RA)-induced differentiation of human embryonal carcinoma cells engages p53. Unexpectedly, RA has been found to activate the transactivation function of p53 in the human embryonal carcinoma cell line, NT2/D1, in a retinoid receptor-dependent manner. A derived RA resistant line, NT2/D1-R1, is deficient in this activity and is co-resistant to cisplatin. This indicates that RA and cisplatin responses may share a common pathway involving p53 in embryonal carcinomas. RA has no effect on p53 steady state protein levels in either line. RA enhances endogenous p53 transactivation activity in NT2/D1 but not NT2/D1-R1 cells. In addition, RA induces transactivation activity of a gal4-p53 fusion protein, suggesting that RA activates p53 independent of increasing p53 levels or sequence-specific DNA binding. This activity is absent in retinoic acid receptor gamma (RARgamma) deficient NT2/D1-R1 cells but can be restored upon co-transfection with specific RARs. Transient transfection of a dominant-negative p53 construct in NT2/D1 cells blocks the RA-mediated transcriptional decline of a differentiation-sensitive reporter plasmid and enhances survival of NT2/D1 cells following cisplatin treatment. Taken together, these findings indicate that RA activates the intrinsic activation function of p53 by a novel mechanism independent of effects on p53 stability or DNA binding and that this activation may be a general mechanism that contributes to RA-mediated G1 arrest. PMID- 11420667 TI - The retinoblastoma-related Rb2/p130 gene is an effector downstream of AP-2 during neural differentiation. AB - Rb2/p130, a member of the Retinoblastoma family of growth and tumour suppressor genes, is extensively implicated in the control of cell cycle and differentiation. The minimal promoter region of Rb2/p130 in T98G human glioblastoma cells was identified and its analysis revealed the presence of a KER1 palindromic sequence able to bind the transcription factor AP-2, a regulatory protein that plays a crucial role in ectodermal differentiation. This KER1 site interacted in vitro with AP-2, and AP-2 overexpression increased Rb2/p130 transcription and translation. We also found that rat PC12 pheochromocytoma cells, when induced to differentiate by NGF, displayed an increase of AP-2 protein levels and of Rb2/p130 transcription and protein levels. AP-2-transfected PC12 cells displayed enhanced transcription and translation of Rb2/p130 and of the cdk inhibitor p21(WAF1/CIP1), a gene known to be under the control of AP-2, but unable by itself to elicit PC12 differentiation. Overexpression of either AP-2 or Rb2/p130 elicited per se cell differentiation in the absence of NGF, while coexpression of AP-2B, a negative regulator of AP-2 transcriptional activity, inhibited only AP-2-induced differentiation. Altogether, these results indicate that Rb2/p130 is a critical effector of AP-2 in sustaining ectodermal differentiation. PMID- 11420668 TI - In situ detection of unexpected patterns of mutant p53 gene expression in non small cell lung cancers. AB - Many solid tumors, including non-small cell lung cancers (NSCLCs), are characterized by heterogenous expression of p53 protein in the neoplastic cells. To analyse the molecular implications of this finding, we examined topographic distribution of p53 mutations using in situ polymerase chain reaction (PCR) in primary NSCLCs, showing distinct patterns of variable p53 overexpression by immunohistochemistry. Unique sets of primers for each mutation were designed, and optimal PCR conditions were determined by standard PCR using DNA from cloned mutants or cell lines established from these tumors. All tumor cell nuclei, regardless of the status of p53 overexpression, demonstrated homogeneous distribution of mutant p53 with specific primers, indicating that only subgroups of the mutated cells overexpressed p53 protein. In situ reverse transcription (RT)-PCR was applied to detect mutant mRNA in the individual tumor cells using specific primers. We found that in each case the distribution of mutant p53 mRNA coincided with that of immunohistochemical overexpression of p53 protein. Our results suggest that the regulation of mutant p53 expression, but not the genotype, is heterogeneous in the neoplastic cells. The topographic genomapping of p53 in NSCLC using in situ PCR provides a novel approach to view molecular mechanisms of lung carcinogenesis. PMID- 11420669 TI - Functional analysis and intracellular localization of p53 modified by SUMO-1. AB - p53 tumor suppressor is a subject of several post-translational modifications, including phosphorylation, ubiquitination and acetylation, which regulate p53 function. A new covalent modification of p53 at lysine 386 by SUMO-1 was recently identified. To elucidate the function of sumoylated p53, we compared the properties of wild type p53 and sumoylation-deficient p53 mutant, K386R. No differences were found between wild type p53 and K386R mutant of p53 in transactivation or growth suppression assays. Moreover, overexpression of SUMO-1 has no effect on p53-regulated transcription. Biochemical fractionation showed that sumoylated p53 is localized in the nucleus and is tightly bound to chromatin structures. p53 and SUMO-1 co-localized in PML nuclear bodies in 293 cells and the nucleoli in MCF7 and HT1080 cells. However, sumoylation-deficient p53 mutant showed a similar pattern of intranuclear localization, suggesting that SUMO-1 does not target p53 to subnuclear structures. These data indicate that SUMO-1 modification of p53 at lysine 386 may not be essential for p53's cellular localization, transcriptional activation, or growth regulation. PMID- 11420670 TI - Rearrangements of minisatellites in the human telomerase reverse transcriptase gene are not correlated with its expression in colon carcinomas. AB - Telomerase activation is crucial in human carcinogenesis. The limiting component of telomerase, the catalytic subunit (hTERT), is undetectable in normal somatic cells but present in most tumor cells, including the earliest stages of colon carcinoma. The mechanisms involved in the differential expression in normal and tumor cells are not understood. In normal cells hTERT expression is shut down by a repressor, and upregulation could be a consequence of cis-acting changes in the hTERT gene, making it resistant to repression. We have identified a polymorphic and a monomorphic minisatellite in the second intron of the hTERT gene, and polymorphic one in intron 6. The polymorphic minisatellite in intron 2 contains binding sites for c-Myc, which has been shown to upregulate hTERT transcription. Screening colon carcinoma DNAs for rearrangements of hTERT minisatellites we detected no changes in 33 samples from tumors, most of which express hTERT. This indicates that size rearrangements of the hTERT minisatellites are not required for telomerase expression in colon carcinomas. Minor changes and one LOH were seen in five tumors. PMID- 11420671 TI - Sustained activation of the Raf/MEK/Erk pathway in response to EGF in stable cell lines expressing the Hepatitis C Virus (HCV) core protein. AB - Chronic hepatitis C virus (HCV) infection is a leading cause of liver cirrhosis and hepatocellular carcinoma (HCC) worldwide. The HCV capside core is a multifunctional protein with regulatory functions that affects transcription and cell growth in vitro and in vivo. Here, we show that both HCV genotype 1a and 3 core proteins activate MEK1 and Erk1/2 MAP kinases and that the costitutive expression of the HCV core results in a high basal activity of Raf1 and MAP/kinase/kinase, as determined by endogenous Raf1 in vitro kinase assay and immunodetection of hyperphosphorylated Erk1 and Erk2 even after a serum starvation. Moreover, the activation of both Erk1/2 and the downstream transcription factor Elk-1 in response to the mitogenic stimulus EGF is significantly prolonged. The sustained response to EGF in cells expressing the HCV core occurs despite a normal induction of the MAP phosphatases MKP regulatory feedback and is likely due to the costitutive activation of Raf-1 activity. The ability of HCV core proteins to directly activate the MAP kinase cascade and to prolong its activity in response to mitogenic stimuli may contribute to the neoplastic transformation of HCV infected liver cells. PMID- 11420672 TI - The role of tetramerization in p53 function. AB - The tumour suppressor gene p53 is extensively studied for its importance in cancer. In its active conformation, p53 is tetrameric and one domain - the tetramerization domain - permits the oligomerization of this protein. Until recently, little attention was given to this domain because, in contrast to the DNA-binding domain, it is not often mutated in cancer. However, various experimental studies have shown evidence that the tetramerization domain is essential for DNA binding, protein-protein interactions, post-translational modifications, and p53 degradation. Moreover, single mutations in the tetramerization domain can inactivate the wild-type protein in a manner similar to that seen with mutations in the DNA-binding domain. Interestingly, the phenotype of several tetramerization domain mutants differs from that observed with DNA-binding domain mutants. In this review, current knowledge about the importance of the tetramerization domain to the function of p53 will be summarized. PMID- 11420673 TI - Specific association of Type I c-Abl with Ran GTPase in lipopolysaccharide mediated differentiation. AB - Each of several isoforms of c-Abl may be involved in different biological functions. Type I c-Abl has been shown to be involved in LPS-induced differentiation and Type IV c-Abl, apoptosis. Ran has recently been shown to be involved in LPS endotoxin signal transduction. Here we show that Type I c-Abl associates with Ran. Formation of this complex is specific, as Ran did not associate with the highly homologous Type IV c-Abl isoform. In non-stimulated lymphoid B cells, Type I c-Abl tyrosine kinase is inactive, whereas Type IV kinase is active. Formation of Type I c-Abl/Ran complex and activation of Type I c-Abl kinase activity are LPS dose-dependent. This complex is detectable in B cells of endotoxin-sensitive inbred mice but absent in B cells of endotoxin resistant mice. These findings therefore suggest that Type I c-Abl and Ran are important targets in lipopolysaccharide-induced biological responses of hematopoietic cells. PMID- 11420674 TI - Different modes and qualities of tyrosine phosphorylation of Fak and Pyk2 during epithelial-mesenchymal transdifferentiation and cell migration: analysis of specific phosphorylation events using site-directed antibodies. AB - Integrin signaling is activated during epithelial-mesenchymal transdifferentiation (EMT) and cell migration, processes serving as models for carcinogenesis. We have shown that paxillin and p130Cas become highly tyrosine phosphorylated during these processes in NMuMG cells. Here, we examined the regulation of Fak and Pyk2, kinases implicated in this phosphorylation. Pyk2 became phosphorylated at the major autophosphorylation site (Tyr-402) and the potential Grb2-binding site (Tyr-881) during EMT. In contrast, phosphorylation of Fak at the corresponding autophosphorylation site (Tyr-397) occurred even in sedentary epithelial cells, whereas phosphorylation at Tyr-407 and Tyr-861 was induced during EMT. During cell migration, these phosphorylation events, except Fak Tyr-397, were augmented further, and phosphorylation of Fak Tyr-577 and the corresponding Pyk2 Tyr-580, both within the kinase activation loops, was also induced. In all cases, phosphorylation of the putative Grb2-binding site in Fak (Tyr-925) was almost undetectable. Although Fak and Pyk2 have several phosphorylation sites in common, Tyr-407 and Tyr-861 are unique to Fak. Our results revealed that Fak and Pyk2 are non-equivalent in the tyrosine phosphorylation events and thereby likely to evoke different downstream signaling cascades during EMT and cell migration of NMuMG cells. We also show that Fak Tyr 397 phosphorylation occurs exclusively at the cytoplasm, but not at focal contacts, in the sedentary epithelial cells. In contrast, all other tyrosine phosphorylated forms of Fak and Pyk2 are predominantly localized to focal adhesions and the cell periphery in motile cells, all colocalized with paxillin and p130Cas. PMID- 11420675 TI - Autocrine and paracrine effects of an ES-cell derived, BCR/ABL-transformed hematopoietic cell line that induces leukemia in mice. AB - During differentiation in vitro, Embryonic Stem (ES) cells generate both primitive erythroid and definitive myeloid lineages in a process that mimics hematopoiesis in the mammalian yolk sac. To investigate leukemic transformation of these embryonic hematopoietic progenitors, we infected differentiating cultures of ES cells with the Chronic Myeloid Leukemia-specific BCR/ABL oncoprotein. Following a period of liquid culture, we isolated two transformed subclones, EB57 and EB67, that retained characteristics of embryonic hematopoietic progenitors and induced a fatal leukemia in mice characterized by massive splenomegaly and granulocytosis. Histopathology of the spleen revealed an abundance of undifferentiated blast-like cells. Investigation of the clonal origins of the granulocytes in the peripheral blood demonstrated that the injected donor cells contributed modestly to the granulocyte population while the majority were host-derived. EB57 secretes IL-3 and unidentified cytokines that can stimulate autocrine and paracrine cell proliferation, presumably accounting for the reactive granulocytosis in diseased mice. These BCR/ABL transformed hematopoietic derivatives of ES cells recapitulate the relationship of BCR/ABL expression to IL-3 production that has been described for primitive hematopoietic progenitors from human CML patients, and illustrates the potential for autocrine and paracrine effects of BCR/ABL-infected cells in murine models. PMID- 11420676 TI - Characterization of germline TP53 splicing mutations and their genetic and functional analysis. AB - Germline TP53 splicing mutations have been described infrequently (>2%) in the literature, however in a series of 40 patients and families identified by our group in which there are germline TP53 mutations, seven affect splicing (18%). The low figure reported in the literature might reflect the method of mutation detection, which in many studies does not include all splice junctions. These data indicate that a significant proportion of TP53 germline mutations are currently unrecognized. We have carried out detailed studies of the effects of the different mutations on splicing, and see distinct variations in the effects of the same mutation in different patients. Furthermore we have identified the usage of a non-consensus splice donor site in four families with an intron 4 splice donor mutation. PMID- 11420677 TI - Paracrine and autocrine effects of fibroblast growth factor-4 in endothelial cells. AB - Recombinant Fibroblast Growth Factor-4 (FGF4) and FGF2 induce extracellular signal-regulated kinase-1/2 activation and DNA synthesis in murine aortic endothelial (MAE) cells. These cells co-express the IIIc/Ig-3 loops and the novel glycosaminoglycan-modified IIIc/Ig-2 loops isoforms of FGF receptor-2 (FGFR2). The affinity of FGF4/FGFR2 interaction is 20-30 times lower than that of FGF2 and is enhanced by heparin. Overexpression of FGF2 or FGF4 cDNA in MAE cells results in a transformed phenotype and increased proliferative capacity, more evident for FGF2 than FGF4 transfectants. Both transfectants induce angiogenesis when applied on the top of the chick embryo chorioallantoic membrane. However, in contrast with FGF2-transfected cells, FGF4 transfectants show a limited capacity to growth under anchorage-independent conditions and lack the ability to invade 3D fibrin gel and to undergo morphogenesis in vitro. Also, they fail to induce hemangiomas when injected into the allantoic sac of the chick embryo. In conclusion, although exogenous FGF2 and FGF4 exert a similar response in MAE cells, significant differences are observed in the biological behavior of FGF4 versus FGF2 transfectants, indicating that the expression of the various members of the FGF family can differently affect the behavior of endothelial cells and, possibly, of other cell types, including tumor cells. PMID- 11420678 TI - N-myc translation is initiated via an internal ribosome entry segment that displays enhanced activity in neuronal cells. AB - Eukaryotic translation can be initiated either by a cap-dependent mechanism or by internal ribosome entry, a process by which ribosomes are directly recruited to structured regions of mRNA upstream of the initiation codon. We analysed the 5' untranslated region (UTR) of the proto-oncogene N-myc, and demonstrated by transfections in a dicistronic vector system that it contains a potent internal ribosome entry segment (IRES). The IRES is similar in length to the c-myc IRES and the activities of these IRESs are comparable in non-neuronal cells. Transfections were also carried out in cell lines derived from neuroblastomas, in which N-myc is expressed, and in a neuronal precursor cell line. In these cells the N-myc IRES is up to seven times more active than that of c-myc, suggesting that neuronal-specific non-canonical trans-acting factors are used by the N-myc but not the c-myc IRES. N-myc expression is increased by gene amplification in many neuroblastomas, but this is the first example of a translational mechanism by which N-myc expression could be further increased. The discovery of an IRES that displays enhanced activity in neuronal cell lines has important potential as a tool for protein expression in neural tissue. PMID- 11420679 TI - Alterations in p53 and E2F-1 function common to immortalized chicken embryo fibroblasts. AB - A number of non-virally and non-chemically immortalized chicken embryo fibroblast (CEF) cells have been established recently in continuous cell culture. All immortal CEF cells tested showed common genetic alterations in the expression patterns of p53 and E2F-1 mRNA and protein which were down- and up-regulated, respectively. The biological effects of differentially regulated p53 and E2F-1 were determined by reporter gene transcriptional activity assays, DNA binding assays, and Northern blot analysis of the expression patterns of down-stream genes. In addition, expression of most of the cyclin genes was up-regulated in immortal CEF cells, which may be associated with the rapid cell division rates and serum-independent growth patterns seen in immortal CEF cells. The telomeric lengths and chromosome integrity were maintained in all immortal CEF cell lines without detectable telomerase activity. Although the functional inactivations of the p53 and Rb regulatory pathways are known to be common events for cellular immortalization, the genetic changes leading to alteration of p53 and E2F-1 function through transcriptional and post-transcriptional regulation seem to be unique in immortal CEF cells. PMID- 11420680 TI - Transcription factors Oct-1 and NF-YA regulate the p53-independent induction of the GADD45 following DNA damage. AB - The p53-regulated GADD45 gene is one of the important players in cellular response to DNA damage, and probably involved in the control of cell cycle checkpoint, apoptosis and DNA repair. There are both the p53-dependent and independent pathways that regulate GADD45 induction. Following ionizing radiation, induction of the GADD45 gene is regulated by p53 through the p53 binding motif located in the third intron of the GADD45 gene. In contrast, GADD45 induction by methyl methanesulfonate (MMS), UV radiation (UV), and medium starvation is independent of p53 status although p53 may contribute to these responses. However, the regulatory elements that control the p53-independent induction of GADD45 remain uncertain. In this report, we have performed detailed analyses to characterize the responsive components that are required for the induction of the GADD45 promoter. We have found that the region between -107 and 62 of the GADD45 promoter is crucial for the induction. Sequence analysis indicates that there are two OCT-1 sites and one CAAT box located in this region. Site-directed mutations of both OCT-1 and CAAT motifs substantially abrogate the induction of the GADD45 promoter by DNA damage. In addition, both Oct-1 protein (binding to OCT-1 site) and NF-YA protein (binding to CAAT box) are induced after cell exposure to DNA damaging agents. Moreover, the Electrophoretic Mobility Shift Assay (EMSA) has demonstrated the direct bindings of Oct-1 and NF-YA proteins to their consensus sequences in the GADD45 promoter. Therefore, these results have presented the novel observation that transcription factors Oct-1 and NF-YA participate in the cellular response to DNA damage and are involved in the regulation of stress-inducible genes. PMID- 11420681 TI - Identification of functional domains involved in BTG1 cell localization. AB - We have previously shown that BTG1 stimulates myoblast differentiation. In addition, this protein displays a major nuclear localization in confluent myoblasts, decreasing during the early steps of differentiation, and is essentially detected in the cytoplasm of mature myotubes. To identify the domains involved in the cellular trafficking of BTG1, we observed the localization of several BTG1 sequences fused to betaGalactosidase. The highly conserved B box among all members of the BTG family induces a significant nuclear localization of the betaGal moiety, enhanced by presence of the BTG1 carboxy-terminal sequence. In addition, a functional Nuclear Export Signal (NES) overlaps the B box. Moreover, presence of the first 43 NH(2)-terminal amino acids reduced the nuclear localization of each chimeric protein tested. Last, the BTG1 amino-terminal domain bears an LxxLL motif favouring nuclear accumulation, and another region encompassing the A box inhibiting nuclear localization. In contrast to a BTG1 mutant exclusively localized in the cytoplasm, transient expression of a mutant displaying a nuclear localization enhanced myoblasts withdrawal from the cell cycle and terminal differentiation, thus mimicking the myogenic influence of BTG1. In conclusion, several regions of BTG1 are implicated in its cellular localization, and BTG1 myogenic activity is induced at the nuclear level. PMID- 11420682 TI - Expression profiling and identification of novel genes in hepatocellular carcinomas. AB - Liver cancer is the fifth most common cancer worldwide and unlike certain other cancers, such as colon cancer, a mutational model has not yet been developed. We have performed gene expression profiling of normal and neoplastic livers in C3H/HeJ mice treated with diethylnitrosamine. Using oligonucleotide microarrays, we compared gene expression in liver tumors to three different states of the normal liver: quiescent adult, regenerating adult, and newborn. Although each comparison revealed hundreds of differentially expressed genes, only 22 genes were found to be deregulated in the tumors in all three comparisons. Three of these genes were examined in human hepatocellular carcinomas and were found to be upregulated. As a second method of analysis, we used Representational Difference Analysis (RDA) to clone mRNA fragments differentially expressed in liver tumors versus regenerating livers. We cloned several novel mRNAs that are differentially regulated in murine liver tumors. Here we report the sequence of a novel cDNA whose expression is upregulated in both murine and human hepatocellular carcinomas. Our results suggest that DEN-treated mice provide an excellent model for human hepatocellular carcinomas. PMID- 11420683 TI - LUCA-15 suppresses CD95-mediated apoptosis in Jurkat T cells. AB - The candidate tumour suppressor gene, LUCA-15, maps to the lung cancer tumour suppressor locus 3p21.3. Overexpression of an alternative RNA splice variant of LUCA-15 has been shown to retard human Jurkat T cell proliferation and to accelerate CD95-mediated apoptosis. An antisense cDNA to the 3'-UTR of this splice variant was able to suppress CD95-mediated apoptosis. Here, we report that overexpression of LUCA-15 itself suppresses CD95-mediated apoptosis in Jurkat cells. This suppression occurs prior to the final execution stage of the CD95 signalling pathway, and is associated with up-regulation of the apoptosis inhibitory protein Bcl-2. LUCA-15 overexpression is also able to inhibit apoptosis induced by the protein kinase inhibitor staurosporine, but is not able to significantly suppress apoptosis mediated by the topoisomerase II inhibitor etoposide. These findings suggest that LUCA-15 is a selective inhibitor of cell death, and confirm the importance of the LUCA-15 genetic locus in the control of apoptosis. PMID- 11420684 TI - p53-interacting protein 53BP2 inhibits clonogenic survival and sensitizes cells to doxorubicin but not paclitaxel-induced apoptosis. AB - 53BP2 was initially identified as a protein interacting with p53 in a yeast two hybrid screen and subsequently shown to enhance p53 transcriptional transactivation and induce apoptosis when transiently overexpressed in cell lines. In order to further study the biologically relevant effects of 53BP2, we have constructed HEK293 stable cell lines where 53BP2 expression can be regulated using an ecdysone inducible expression system. Our results indicate that the response of cells is dependent on the amount of 53BP2 that is expressed. High levels of 53BP2 expression (>or=140-fold above endogenous) impede cell cycle progression and induce apoptosis. Lower levels of 53BP2 expression (6-11-fold above endogenous) suppress colony formation but do not lead to detectable perturbations in the cell cycle or apoptosis. Lower levels of 53BP2 expression sensitized cells to apoptosis induced by DNA damaging chemotherapy agents doxorubicin, ara-C and VP16, but not microtubule active agents paclitaxel and vinblastine. Our results demonstrate that high levels of 53BP2 expression have profound biological effects ultimately leading to apoptosis, whereas lower levels of 53BP2 expression have more subtle effects on growth and sensitize cells to some chemotherapy agents. PMID- 11420685 TI - Tumor suppressor protein VHL is induced at high cell density and mediates contact inhibition of cell growth. AB - In spite of the general recognition of von Hippel-Lindau (VHL) as a tumor suppressor gene, the physiological and pathological importance of VHL protein in cell growth regulation and tumorigenesis remains unclear. Here we show that in normal human renal proximal tubule epithelial cells (RPTEC), the steady-state amount of VHL protein is strictly regulated by cell density. The cellular VHL content is more than 100-fold higher in dense cultures than in sparse cultures. The increase in VHL protein at high cell density was also observed for NIH3T3 fibroblasts, suggesting the generality of the phenomenon. The growth rates of renal cell carcinoma cells lacking an intact VHL gene and their derivatives with wild-type or mutant VHL expression vector do not differ significantly when they are growing in log-phase. Importantly, however, there is a difference when they reach confluency: cells lacking wild-type VHL grew continuously, while cells expressing exogenous VHL protein showed relatively limited cell growth. Using an ecdysone-inducible VHL expressing cell line, we also show that the growth inhibition at high cell density can be released by attenuating the VHL expression. Taken together, we propose that VHL protein functions as a growth suppressor at high cell density, and this might be the basis of the tumor suppressor function of VHL. PMID- 11420686 TI - Exploitation of a non-apoptotic caspase to regulate the abundance of the cdkI p27(KIP1) in transformed lymphoid cells. AB - Expression of the cyclin dependent kinase inhibitor p27(KIP1) is intimately linked to the control of proliferation, and is itself regulated by transcription, translation, phosphorylation, protein stability or sequestration. p27(KIP1) is also regulated during apoptosis; cleavage occurs at DPSD(139)S and ESQD(108)V, by a sub-set of Z-VAD-fmk-sensitive caspases. We have identified a related but distinct mechanism that regulates p27(KIP1) in proliferating lymphoid cell lines. In a B-lymphoid cell line (BJAB), the abundance of p27(KIP1) oscillates inversely to proliferation; loss of full-length p27(KIP1) correlates with the appearance of a truncated version corresponding to cleavage at DPSD(139)S. A direct correlation exists between the appearance of truncated p27(KIP1) and the presence of an activity able to cleave peptides representing DPSD(139)S and a caspase-8 substrate (Ac-IETD-AMC) in vitro. This activity is inhibited by Ac-IETD-CHO but not Z-VAD-fmk in vitro. Furthermore a requirement for caspase-8 has been excluded. The activity differs from the apoptosis related p27(KIP1)-cleaving activity; indeed few cells undergoing apoptosis are present in the population of proliferating cells. The activity is further distinguished by its inability to cleave a peptide based on ESQD(108)V in vitro, together with the lack of a corresponding cleavage product in vivo. Inhibition of the caspase activity in vivo promotes an accumulation of full length p27(KIP1), as well as a decrease in cell proliferation. Together these studies highlight the importance of non apoptotic caspases in regulating p27(KIP1) in transformed lymphoid cells. PMID- 11420687 TI - Overexpression of caspase-3 restores sensitivity for drug-induced apoptosis in breast cancer cell lines with acquired drug resistance. AB - In this study, we asked whether overexpression of caspase-3, a central downstream executioner of apoptotic pathways, might sensitize breast cancer cells with acquired drug resistance (MT1/ADR) to drug-induced apoptosis. As control, we employed caspase-3 negative and caspase-3-transfected MCF-7 cells. Whereas mock transfected MCF-7 cells were resistant to epirubicin, etoposide and paclitaxel (taxol), the same drugs led to breakdown of nuclear DNA in caspase-3-transfected MCF-7 cells. MT1/ADR cells express low levels of wild type caspase-3 but show defective caspase activation and apoptosis upon drug exposure. These cells also display a less efficient activation of the mitochondrial permeability transition. Caspase-3-transfected MT1/ADR clones showed a 2.8-fold increase in the protein level and a 3.7-fold higher specific enzyme activity. Procaspase-3 overexpression was not toxic and did not affect background apoptosis. Interestingly, procaspase 3-transfected MT1/ADR cells were more sensitive to cytotoxic drugs as compared with vector-transfected controls and DNA fragmentation nearly reached the levels of the original drug sensitive MT1 cells. Thus, overexpression of caspase-3 enhances chemosensitivity especially in situations where activation of the mitochondrial apoptosome is disturbed. PMID- 11420688 TI - Down-regulation of MET, the receptor for hepatocyte growth factor. AB - The ligand-dependent degradation of activated tyrosine kinase receptors provides a means by which mitogenic signalling can be attenuated. In many cell types the ligand-dependent degradation of the tyrosine kinase receptor Met is completely dependent on the activity of the 26S proteasome (Jeffers et al., 1997b). We now show that degradation also requires trafficking to late endosomal compartments and the activity of acid dependent proteases as determined by the effects of a dominant negative form of dynamin (K44A) and a vacuolar-ATPase inhibitor, concanamycin. We show that in the presence of the proteasome inhibitor lactacystin, Met fails to redistribute from the plasma membrane to intracellular compartments. This observation is most consistent with the interpretation that proteasome activity is required for Met internalization and only indirectly for its degradation. PMID- 11420689 TI - AP-1 blockade inhibits the growth of normal and malignant breast cells. AB - We have previously demonstrated that basal AP-1 transcriptional activity is high in normal human mammary epithelial cells, intermediate in immortal breast cells, and relatively low in breast cancer cells. In this study we investigated whether differences in AP-1 transcriptional activity reflect differences in breast cells' dependence on AP-1 for proliferation. The cJun dominant negative, TAM-67, was used to determine the effect of AP-1 blockade on the growth of normal, immortal and malignant breast cells. We first showed that TAM-67 inhibits AP-1 activity in normal and malignant breast cells. We then determined whether this AP-1 inhibitor affected colony forming efficiency of the immortalized and malignant breast cells. The AP-1 inhibitor reduced colony formation of immortal breast cells by over 50% (by 58% in 184B5 cells and 62% in MCF10A cells), and reduced colony formation in the breast cancer cell line MCF7 by 43%, but did not reduce colony formation in the other breast cancer cell lines (T47D, MDA MB231 and MDA MB 435). We also determined the effect of AP-1 blockade on the growth of normal breast cells using a single cell proliferation assay. Using this assay, the growth of normal breast cells was extremely sensitive to AP-1 blockade, while immortal breast cells were moderately sensitive. We next directly tested the effect of TAM 67 expression on the growth of MCF7 breast cancer cells, using cells stably transfected with TAM-67 under the control of a doxycycline-inducible promoter. Upon induction, TAM-67 was expressed and AP-1 activity was inhibited in these cells. We then measured the growth of these cells in the presence or absence of TAM-67. The results of these studies show that the growth of MCF7 cells was suppressed by the AP-1 inhibitor, TAM-67. These results demonstrate that normal and immortalized breast cells, and some breast cancer cells (such as MCF7), require AP-1 to transduce proliferative signals, while other breast cancer cells (such as T47D, MDA MB 231 and MDA MB 435) do not. These studies suggest that the AP-1 transcription factor is a potential target for future agents for the prevention or treatment of breast cancer. PMID- 11420690 TI - Dolichol-phosphate-mannose-3 (DPM3)/prostin-1 is a novel phospholipase C-gamma regulated gene negatively associated with prostate tumor invasion. AB - The most ominous development in tumor progression is the transition to an invasive and metastatic phenotype. Little is known, however, about the molecular alterations that cause a tumor to become invasive. In view of this, we have used microarray expression analysis to evaluate the expression profiles of a unique panel of human DU145 prostate cancer sublines that vary in their invasive potential. The three DU145 sublines expressed epidermal growth factor (EGF) receptors that differed in their ability to activate phospholipase C-gamma (PLC gamma). Three-way analyses yielded 11 genes out of 4608 genes screened that associated directly or inversely with invasive potential. The gene whose expression correlated most strongly with lack of invasion was identified as a potential invasion suppressor and called prostin-1. Pharmacological inhibition of PLC gamma (U73122) confirmed that PLC gamma signaling suppressed prostin-1 in that U73122 treatment caused induction of prostin-1 in PLC gamma competent cells. The prostin-1 gene, conserved through phylogeny, is induced by androgen in LNCaP cells and encodes a 92 amino acid protein. The protein shares no extensive homologies with other known genes, yet was recently identified as a small stabilizer subunit of the dolichol-phosphate-mannose (DPM) synthase complex. That DPM3/prostin-1 might suppress tumor progression was supported by the finding that exogenous expression in COS cells leads to apoptosis. These findings support the use of model cell lines to identify putative tumor suppressors and promoters. PMID- 11420691 TI - FGF-8b increases angiogenic capacity and tumor growth of androgen-regulated S115 breast cancer cells. AB - Fibroblast growth factor 8 (FGF-8) is a secreted heparin-binding protein, which has transforming potential. Alternative splicing of the mouse Fgf-8 gene potentially codes for eight protein isoforms (a-h) which differ in their transforming capacity in transfected cells. S115 mouse mammary tumor cells express a transformed phenotype and secrete FGF-8 in an androgen-dependent manner. In order to study the role of FGF-8 isoforms in the induction of transformed phenotype of breast cancer cells, we over-expressed FGF-8 isoforms a, b and e in S115 cells. Over-expression of FGF-8b, but not FGF-8a or FGF-8e, induced androgen and anchorage independent growth of S115 cells. FGF-8b transfected S115 cells formed rapidly growing tumors with increased vascularization when injected s.c. into nude mice. FGF-8a also slightly increased tumor growth and probably tumor vascularization but FGF-8e was not found to have any effects. The angiogenic activity of FGF-8b and heparin-binding growth factor fraction (HBGF) of S115 cell conditioned media was tested in in vitro and in vivo models for angiogenesis using immortomouse brain capillary endothelial cells (IBEC) and chorion allantoic membrane (CAM) assays. Recombinant FGF-8b protein was able to stimulate proliferation, migration, and vessel-like tube formation of IBECs. In addition, stimulatory effect of S115-HBGF on IBE cell proliferation was evident. A positive angiogenic response to FGF-8b was also seen in CAM assay. The results demonstrate that the expression of Fgf-8b is able to promote vessel formation. Angiogenic capacity probably markedly contributes to the ability of FGF-8b to increase tumor growth of androgen-regulated S115 mouse breast cancer cells. PMID- 11420692 TI - Inhibition of the NF-kappa B transcription factor increases Bax expression in cancer cell lines. AB - The NF-kappa B transcription factor has been shown to inhibit apoptosis in several experimental systems. We therefore investigated whether the expression of the Bax proapoptotic protein could be influenced by NF-kappa B activity. Increased Bax protein expression was detected in HCT116, OVCAR-3 and MCF7 cells stably expressing a mutated unresponsive I kappa B-alpha inhibitory protein that blocks NF-kappa B activity. Northern blots showed that bax mRNA expression was increased as a consequence of mutated I kappa B-alpha expression in HCT116 cells. A careful examination of the human bax gene promoter sequence showed three putative binding sites for NF-kappa B, and the kappa B2 site at position -687 could indeed bind NF-kappa B complexes in vitro. Transient transfection of a bax promoter luciferase construct in HCT116 cells showed that NF-kappa B proteins could partially inhibit the transactivation of the bax promoter by p53. Mutations or deletions of the kappa B sites, including kappa B2, indicated that this NF kappa B-dependent inhibitory effect did not require NF-kappa B DNA-binding, and was thus an indirect effect. However, cotransfection of expression vectors for several known cofactors failed to identify a competition between p53 and NF-kappa B for a transcription coactivator. Our findings thus demonstrate for the first time that NF-kappa B regulates, through an indirect pathway, the bax gene expression. PMID- 11420693 TI - Myc down-regulation induces apoptosis in M14 melanoma cells by increasing p27(kip1) levels. AB - In recent years, increasing evidence indicated the importance of a deregulated c myc gene in the melanoma pathogenesis. We have previously demonstrated that treatment of melanoma cells with c-myc antisense oligodeoxynucleotides can inhibit cell proliferation and activate apoptosis. To gain insight into the mechanisms activated by Myc down-regulation, we have now developed an experimental model that allows modulating Myc protein expression in melanoma cells. This was achieved by originating stable melanoma cell clones expressing ecdysone-inducible c-myc antisense RNA. We show that the induction of c-myc antisense RNA in M14 melanoma cells leads to an inhibition of cell proliferation characterized by accumulation of cells in the G(1) phase of the cell cycle (up to 80%) and activation of apoptosis (50%). These data are associated with an increase of p27(kip1) levels and a significant reduction of the cdk2-associated kinase activity. In addition, we show that an ectopic overexpression of p27(kip1) in this experimental model can enhance the apoptotic rate. Our results indicate that down-regulation of Myc protein induces a G(1) arrest and activates apoptosis by increasing p27(kip1) content in melanoma cells, that are known to be defective for the p16-cyclinD/cdk4-pRb G(1) checkpoint. This is particularly relevant for identifying new therapeutic strategies based on the re-establishment of the apoptotic pathways in cancer cells. PMID- 11420694 TI - Transcriptional activation of TRADD mediates p53-independent radiation-induced apoptosis of glioma cells. AB - Survival of patients with Glioblastoma Multiforme (GM), a highly malignant brain tumor, remains poor despite concerted efforts to improve therapy. The median survival of patients with GM has remained approximately 1 year regardless of the therapeutic approach. Since radiation therapy is the most effective adjuvant therapy for GM and nearly half of GM tumors harbor p53 mutations, we sought to identify genes that mediate p53-independent apoptosis of GM cells in response to ionizing radiation. Using broad-scale gene expression analysis we found that following radiation treatment, TRADD expression was induced in a uniquely radiosensitive GM cell line but not in radioresistant GM cell lines. TRADD over expression killed GM cells and activated NF-kappa B. We found that blocking the TRADD-mediated pathway using a dominant-negative mutant of FADD (FADD-DN) enhanced radiation resistance of GM cells, as reflected in both susceptibility to apoptosis and clonogenic survival following irradiation. Conversely, stable expression of exogenous TRADD enhanced radiation-induced apoptosis of GM cell lines, reflecting the biological significance of TRADD regulation in p53 independent apoptosis. These findings generate interest in utilizing TRADD in gene therapy for GM tumors, particularly in light of its dual function of directly inducing rapid apoptosis and sensitizing GM cells to standard anti neoplastic therapy. PMID- 11420695 TI - Bcl-2 oncoprotein protects the human prostatic carcinoma cell line PC3 from TRAIL mediated apoptosis. AB - The role of Bcl-2 in TRAIL-induced apoptosis has been investigated in lymphoid cells. Here we show that the human prostatic carcinoma cell line PC3 was sensitive to TRAIL treatment whereas PC3 overexpressing of Bcl-2 was resistant. TRAIL receptors ligation in PC3 activated caspases -2, -3, -7, -8, and -9, induced Bid processing, dissipation of mitochondrial transmembrane potential (Delta Psi(m)), and cytochrome c release. We have detected caspases -8 and -3 only in the cytosolic fraction of cells, but caspases -2, -7, and -9 were found both in cytosolic and mitochondrial fractions. Bcl-2 overexpression did not affect caspase-8 activation although it did change the processing pattern of caspase-3. At the same time, Bcl-2 overexpression inhibited the activation of mitochondrial localized caspases -2, -7, and -9. Bcl-2 also abrogated TRAIL induced cytochrome c release and dissipation of Delta Psi(m). These findings suggest that TRAIL-induced apoptosis in the epithelial cell line PC3 depends both on mitochondrial integrity and caspase activation. PMID- 11420696 TI - The TES gene at 7q31.1 is methylated in tumours and encodes a novel growth suppressing LIM domain protein. AB - Many studies suggest that a multi-tissue tumour suppressor gene is located at human chromosome 7q31.1. We have cloned and characterized a novel gene at this locus. The TES gene lies within the minimal region of overlap of several LOH studies and appears to possess the properties of a tumour suppressor. TES is widely expressed and is predicted to encode a protein of 421 amino acids, with three C-terminal LIM domains. Mutation analysis of the coding TES exons in 21 human tumour-derived cell lines revealed the presence of a frameshift mutation in one allele in the breast cancer cell line ZR-75. Methylation of the CpG island at the 5' end of TES appears to be a remarkably frequent finding, occurring in seven out of 10 ovarian carcinomas and in each of the 30 tumour-derived cell lines tested. Moreover, forced expression of TES in HeLa or OVCAR5 cells, resulted in a profound reduction in growth potential, as determined by the colony formation assay. We believe that TES is a tumour suppressor gene that is inactivated primarily by transcriptional silencing resulting from CpG island methylation. PMID- 11420697 TI - Global analysis of differential gene expression after transformation with the v-H ras oncogene in a murine tumor model. AB - Mouse PB-3c mast cells stably transfected with the v-H-ras oncogene induce tumor formation in vivo when implanted into mice. Such tumor cells are characterized by an autocrine IL-3 loop. DNA microarrays allow simultaneous transcript imaging of several thousand genes and the technique was applied in this tumor model to analyse gene expression following malignant transformation. Using three independent tumor lines derived from the same precursor the expression of about 400 out of 11 000 genes was modulated in each tumor. A subset of only 75 genes (0.68%) is shared and up- or downregulated in all three lines. A significant portion of this gene pool possesses functions related to tumorigenesis such as cell adhesion, signaling or transcriptional regulation. Apart from a number of expressed sequence tags (EST's) we find downregulation of four interferon inducible genes in the tumor lines. Finally, when we extrapolate our data to the complete mouse genome, we estimate that about 500 genes are differentially expressed in tumor cells compared to the precursor cell PB-3c. PMID- 11420698 TI - p53 contains a DNA break-binding motif similar to the functional part of BRCT related region of Rb. AB - The BRCT regions are two repeating structures at BRCA1 carboxyl-terminus and are ubiquitous in some proteins involved in DNA repair and cell cycle checkpoints. It was shown that BRCTs of TopBP1, BRCA1, and BRCT-Ws of Rb bound DNA ends and breaks. We indicate here that the C-terminus of p53 tumor suppressor contains a DNA binding motif (residues 327-333 in human), whose features are similar to those of the part of BRCT-W in Rb with DNA binding activity. The short motif was required for the gel retardation activity of DNA fragments, since residues 311 333 showed the activity while residues 311-326 showed no activity. Significant numbers of total p53 and its fragments with the motif formed multimerizing complexes and associated with DNA ends and breaks. These results suggest the common presence of DNA binding motifs that can recognize DNA ends or damages in major tumor suppressors, Rb, BRCA1 and p53. The oncogenic activity of p53 C terminus (residues 311-393) required both the DNA damage recognition motif and the repair enzyme-associating domain. PMID- 11420699 TI - Protein kinases required for segregation of vimentin filaments in mitotic process. AB - Vimentin, one of type III intermediate filament (IF) proteins, is expressed not only in mesenchymal cells but also in most types of tumor cells. In the present study, we introduced several types of vimentin mutated at putative phosphorylation sites in its amino-terminal head domain into type III IF-negative T24 cells. Site-specific mutation induced the formation of an unusually long bridge-like IF structure between the unseparated daughter cells, although these mutants formed the filament network similar to wild type in interphase cells. Together with sites phosphorylated by Rho-kinase and protein kinase C (PKC), vimentin-Ser72, which can not be phosphorylated by any known vimentin kinase, was one of the mutation sites essential for this phenotype. We further demonstrated that vimentin-Ser72 was phosphorylated specifically at the cleavage furrow during cytokinesis. These observations suggest the existence of a novel protein kinase responsible for vimentin filament separation through the cleavage furrow-specific vimentin phosphorylation. We propose that Rho-kinase, PKC, and an unidentified vimentin-Ser72 kinase may play important roles in vimentin filament separation during cytokinesis. PMID- 11420700 TI - Defective caspase-3 relocalization in non-small cell lung carcinoma. AB - Many anticancer drugs exert their cytotoxicity through DNA damage and induction of apoptosis. Small cell lung carcinoma (SCLC) and non-small cell lung carcinoma (NSCLC) have different sensitivity to treatment with radiation and chemotherapeutic agents with SCLC being more sensitive than NSCLC both in vitro and in vivo. This difference might be related to the different susceptibility of small and non-small cell lung carcinoma to undergo apoptosis. The aim of this study was to investigate if deficiencies in the apoptotic pathways can explain the intrinsic resistance of NSCLC to anti-cancer treatment. Three different triggers were used to induce apoptosis. Etoposide and gamma-radiation, which are important parts of clinical lung cancer treatment, induce DNA-damage, whereas Fas ligation induces receptor-mediated apoptotic pathways. NSCLC cells were cross resistant to all treatments, whereas SCLC cells, which do not express pro-caspase 8, were resistant to alphaFas-, but not to DNA-damage-induced apoptosis. Cytochrome c release, activation of caspase-9 and the executioner caspase-3 were observed in both types of lung cancer cells. However, cleavage of known nuclear substrates for caspase-3, such as PARP and DFF45/ICAD, was documented only in the sensitive SCLC cells but not in the resistant NSCLC cells. Moreover, relocalization of active caspase-3 from the cytosol into the nucleus upon treatment was observed only in the SCLC cell line. These results indicate that the inhibition of apoptosis in NSCLC occurs downstream of mitochondrial changes and caspase activation, and upstream of nuclear events. PMID- 11420701 TI - Accumulation of high levels of the p53 and p130 growth-suppressing proteins in cell lines stably over-expressing cyclin-dependent kinase 6 (cdk6). AB - Cyclin-dependent kinase 6(cdk6) is present in randomly proliferating cultures of 3T3 cells but has little detectable enzymatic activity. Significant activity is detected only during a short period in early G1 phase. To examine the possible functions of cdk6 in 3T3 cells, lines stably over-expressing cdk6 were constructed and compared to normal 3T3 cells or cell lines with reduced cdk6 levels due to expression of a dominant-negative form of the protein. Over expression of cdk6 in cells, which led to high levels of activity even in proliferating cultures, had dramatic effects. Cell lines stably over-expressing wild-type cdk6 had a markedly reduced growth rate compared to parental 3T3 cells or lines expressing a dominant-negative form of cdk6. They also over-produced the p53 and p130 proteins and had increased sensitivity to UV-irradiation. Irradiation resulted in accumulation of the Bax protein and rapid cell death. Levels of p53 and p130 proteins were down-regulated and the growth rate of the cells was increased by introduction of the dominant-negative form of cdk6 into cells over-expressing cdk6, indicating that cdk6 is involved in the overproduction of p53 and p130. The results suggest that cdk6, through regulation of growth-suppressing molecules, may play a role in halting cellular growth when proliferation is inappropriate. PMID- 11420702 TI - Biased distribution of chromosomal breakpoints involving the MLL gene in infants versus children and adults with t(4;11) ALL. AB - Derivative chromosomes of 40 patients diagnosed with t(4;11) acute lymphoblastic leukemia (ALL) were analysed on the genomic DNA level. Chromosomal breakpoints were identified in most cases within the known breakpoint cluster regions of the involved MLL and AF4 genes. Due to our current knowledge of the primary DNA sequences of both breakpoint cluster regions, specific features were identified at the chromosomal fusion sites, including deletions, inversions and duplications of parental DNA sequences. After separation of all t(4;11) leukemia patients into two age classes (below and above 1 year of age), the analysis of chromosomal fusion sites revealed significant differences in the distribution of chromosomal breakpoints and led to the definition of two hotspot areas within the MLL breakpoint cluster region. This may point to the possibility of different age linked mechanisms that were leading to t(4;11) chromosomal translocations. PMID- 11420703 TI - Mmip-2/Rnf-17 enhances c-Myc function and regulates some target genes in common with glucocorticoid hormones. AB - Members of the Mad family of basic-helix-loop-helix-leucine zipper proteins inhibit the transcriptional activity of the c-Myc oncoprotein. Mmip-2/Rnf-17 is a RING-finger protein that interacts with all four known Mad proteins, redistributes them to the cytoplasm, and thus enhances c-Myc function. We generated cell lines in which Mmip-2/Rnf-17 was rendered glucocorticoid (GC) inducible. Stable expression of Mmip-/Rnf-17 resulted in the expected transport of the most abundant endogenous mad protein, Mxi1, to the cytoplasm. Compensatory increases in Mxi1 and Mad3 transcripts, similar to those previously described in Mad1 null hematopoietic cells, were also seen. Mmip-2/Rnf-17 also sensitized cells to several different pro-apoptotic stimuli and regulated a subset of c-Myc target genes. Unexpectedly, some of these genes were also found to be modulated solely by GCs. Thus, the inhibition of Mad proteins by Mmip-2/Rnf-17 modulates c Myc function by enhancing its ability to regulate a subset of its potential target genes. Our results also identify a previously unrecognized overlap between genes regulated by c-Myc- and GCs and provide a potential molecular basis for their regulation of common cellular functions. PMID- 11420704 TI - Sequential two-step cleavage of the retinoblastoma protein by caspase-3/-7 during etoposide-induced apoptosis. AB - During cellular apoptosis, retinoblastoma protein (RB) is subjected to cleavage near the carboxyl terminus by a caspase-3-like protease. In addition, an heretofore unidentified protease cleaves RB internally, generating fragments of 68 and 48 kDa. Internal cleavage abrogates the ability of RB to associate with E2F. To investigate the mechanism of RB internal cleavage, we developed and employed an in vitro cleavage assay. Incubation of in vitro translated (35)S-RB with apoptotic cell extracts led to RB cleavage at the C-terminus, followed by internal cleavage. The caspase peptide inhibitors z-VAD-FMK or z-DEVD-FMK blocked both cleavage events. Rapid C-terminal and internal cleavage were also observed when recombinant caspase-3 was added to (35)S-RB. Moreover, when caspase-3 was added to nonapoptotic cell extract, efficient internal cleavage of cellular RB was observed. Caspase-mediated internal cleavage occurred following RB residue aspartate(349) in the sequence DSID(349). This sequence is consistent with a DXXD recognition motif for caspase-3-like enzymes. Interestingly, we also observed RB internal cleavage in caspase-3-deficient MCF-7 cells, indicating that other caspases are capable of cleaving RB internally. Indeed, caspase-7, a member of the caspase-3 subfamily, was found to cleave (35)S-RB at both the carboxyl terminus, and following aspartate(349). By contrast, caspases that are not members of the caspase-3 subfamily failed to cleave RB. Taken together, our findings demonstrate that during apoptosis, a caspase-3-like protease is responsible for degradation and functional inactivation of RB by cleaving the protein internally following aspartate(349). PMID- 11420705 TI - Indole-3-carbinol (I3C) induced cell growth inhibition, G1 cell cycle arrest and apoptosis in prostate cancer cells. AB - Prostate cancer is one of the most common cancers in men and it is the second leading cause of cancer related death in men in the United States. Recent dietary and epidemiological studies have suggested the benefit of dietary intake of fruits and vegetables in lowering the incidence of prostate cancer. A diet rich in fruits and vegetables provides phytochemicals, particularly indole-3-carbinol (I3C), which may be responsible for the prevention of many types of cancer, including hormone-related cancers such as prostate. Studies to elucidate the role and the molecular mechanism(s) of action of I3C in prostate cancer, however, have not been conducted. In the current study, we investigated whether I3C had any effect against prostate cancer cells and, if so, attempts were made to identify the potential molecular mechanism(s) by which I3C elicits its biological effects on prostate cancer cells. Here we report for the first time that I3C inhibits the growth of PC-3 prostate cancer cells. Induction of G1 cell cycle arrest was also observed in PC-3 cells treated with I3C, which may be due to the observed effects of I3C in the up-regulation of p21(WAF1) and p27(Kip1) CDK inhibitors, followed by their association with cyclin D1 and E and down-regulation of CDK6 protein kinase levels and activity. The induction of p21(WAF1) appears to be transcriptionally upregulated and independent of the p53 responsive element. In addition, I3C inhibited the hyperpohosphorylation of the Retinoblastoma (Rb) protein in PC-3 cells. Induction of apoptosis was also observed in this cell line when treated with I3C, as measured by DNA laddering and poly (ADP-ribose) polymersae (PARP) cleavage. We also found an up-regulation of Bax, and down regulation of Bcl-2 in I3C-treated cells. These effects may also be mediated by the down-regulation of NF-kappaB observed in I3C treated PC-3 cells. From these results, we conclude that I3C inhibits the growth of PC-3 prostate cancer cells by inducing G1 cell cycle arrest leading to apoptosis, and regulates the expression of apoptosis-related genes. These findings suggest that I3C may be an effective chemopreventive or therapeutic agent against prostate cancer. PMID- 11420706 TI - Adrenomedullin inhibits hypoxic cell death by upregulation of Bcl-2 in endometrial cancer cells: a possible promotion mechanism for tumour growth. AB - Regions of hypoxia are a common feature of solid tumours. When tumour cells are exposed to hypoxic stress, transcription of a battery of genes is initiated. The angiogenic factor adrenomedullin (ADM) is a hypoxia regulated gene. ADM is thought to act through the G protein-coupled receptor calcitonin receptor-like receptor (CRLR), with specificity being conferred by the receptor associated modifying protein 2 (RAMP2). Here we report for the first time that ADM treated or stably transfected Ishikawa cells overexpressing ADM show increased resistance to hypoxia induced apoptosis. These cells also show an upregulation of the oncoprotein Bcl-2, which is protective against hypoxic cell death when transiently transfected into Ishikawa cells. Since Ishikawa cells express the putative ADM-receptor CRLR-RAMP2 the production and secretion of ADM with the consecutive upregulation of Bcl-2 could establish an autocrine/paracrine mechanism rescuing malignant cells from hypoxic cell death. These results, taken together with our previous findings that ADM is an angiogenic factor which is upregulated by the nonsteroidal antiestrogen tamoxifen (TAM) in endometrial cells, implicate this peptide as a promoter of tumour growth and a possible target for anticancer strategies. PMID- 11420707 TI - Loss of p53 tumor suppressor function is required for in vivo progression of Friend erythroleukemia. AB - A role for p53 in the in vivo progression of Friend virus-induced erythroleukemia has been suggested but not clearly defined. We developed a Friend virus sensitive, p53-deficient mouse model to directly address the role of p53 in Friend erythroleukemia. When infected with the polycythemia-inducing strain of Friend virus (FVP), p53 null mice exhibited accelerated progression to erythroleukemia and accelerated death following diagnosis when compared to wild type mice. Confirmation that p53 mutations were required for disease progression was provided by sequence analysis of p53 transcripts in leukemic wild type and heterozygous mice. All transcripts evaluated had point mutations, deletions or insertions in the p53 gene. The ability to grow tumor colonies in vitro and derive cell lines was enhanced in FVP-infected p53 null animals. Although PU.1 oncogene overexpression is a common mutation observed in cell lines derived from Friend virus-infected p53 wild type mice, it was not a universal finding in cell lines derived from p53 null animals. Our data conclusively demonstrate that loss of p53 function is a requirement for progression of Friend erythroleukemia in vivo. Further, the data demonstrate that erythroleukemias arising in Friend virus infected p53 null mice are biologically and genetically distinct from those that occur in wild type animals, suggesting that the temporal order of PU.1 and p53 mutations is an important parameter in the pathogenesis of leukemic development. PMID- 11420708 TI - Negative regulation of G(1)/S transition by the candidate bladder tumour suppressor gene DBCCR1. AB - Deletion of all or part of chromosome 9q is the most common genetic alteration in all stages and grades of bladder cancer. DBCCR1 (deleted in bladder cancer chromosome region candidate 1) maps to the chromosome region 9q32-33, a candidate tumour suppressor locus for bladder cancer. Although no mutations of DBCCR1 have been detected in bladder tumours, expression of DBCCR1 is silenced by promoter hypermethylation in 50% of bladder cancer cell lines analysed. Here we sought to provide functional evidence to authenticate DBCCR1 as a tumour suppressor using gene-transfer methods. Exogenous expression of DBCCR1 protein or an HA epitope tagged fusion protein, HA-DBCCR1 in NIH3T3 cells and human bladder tumour cell lines resulted in suppression of proliferation. Cell cycle analyses in NIH3T3 cells revealed that DBCCR1-mediated growth inhibition was due to an increase in the number of cells in the G(1) phase of the cell cycle. The levels of apoptosis were not altered. These results demonstrate a role for DBCCR1 in cell cycle control, thereby supporting the hypothesis that this is the tumour suppressor gene targeted by 9q32-33 deletion in bladder cancer. PMID- 11420709 TI - Structural and functional analysis of a chimeric protein COL1A1-PDGFB generated by the translocation t(17;22)(q22;q13.1) in Dermatofibrosarcoma protuberans (DP). AB - Dermatofibrosarcoma protuberans (DP), an infiltrative skin tumour of intermediate malignancy, presents specific cytogenetic features such as reciprocal translocations t(17;22)(q22;q13.1) or supernumerary ring chromosomes derived from t(17;22). We have previously shown that both rings and translocated chromosomes derived from t(17;22) presented the same molecular rearrangement with fusion of the COL1A1 gene on chromosome 17 to the PDGFB gene on chromosome 22. To study the structure and function of the COL1A1-PDGFB chimeric protein, we used a tumour derived chimeric COL1A1-PDGFB cDNA to perform stable and transient transfections in the Chinese hamster lung fibroblastic cell line PS200 and the human epithelial cell line HEK293. We demonstrated that the stably transfected clones that expressed the COL1A1-PDGFB chimeric protein became growth factors independent and tumorigenic in nude mice. In addition, COL1A1-PDGFB transfected cell supernatants significantly stimulated fibroblastic cell growth, through the activation of the PDGFB receptor pathway. By using anti-PDGFB and specific anti-COL1A1-PDGFB antibodies, we showed that, similar to PDGFB, the COL1A1-PDGFB chimeric proteins are processed in transfected cells into mature PDGFB dimers. These results strongly suggest that the COL1A1-PDGFB chimeric gene expression associated with DP, induces tumours formation through production of mature PDGFB, in an autocrine or paracrine way. Strikingly, mutagenesis experiments indicated that uncleaved COL1A1-PDGFB forms are mitogenic and therefore could contribute, as well as mature PDGFB, to the establishment of a transformed phenotype. PMID- 11420710 TI - A novel germline 1.8-kb deletion of hMLH1 mimicking alternative splicing: a founder mutation in the Chinese population. AB - We have previously reported that there is a high incidence of microsatellite instability (MSI) and germline mismatch repair gene mutation in colorectal cancer arising from young Hong Kong Chinese. Most of the germline mutations involve hMSH2, which is different from the mutation spectrum in the Western population. It is well known that alternative splicing is common in hMLH1, which complicates RNA based mutation detection methods. In contrast, large deletions in hMLH1, commonly observed in some ethnic groups, tend to escape detection by exon-by-exon direct DNA sequencing. Here we report the detection of a novel germline 1.8 kb deletion involving exon 11 of hMLH1 in a local hereditary non-polyposis colorectal cancer family. This mutation generates a mRNA transcript with deletion of exons 10-11, which is indistinguishable from one of the most common and predominant hMLH1 splice variants. A diagnostic test based on PCR of the breakpoint region led to the identification of an additional young colorectal cancer patient with this mutation. Haplotype analysis suggests that they may share a common ancestral mutation. Our results caution investigators in the interpretation of alternative splicing and have important implications for the design of hMLH1 mutation detection strategy in the Chinese population. PMID- 11420711 TI - A p53 dose-response relationship for sensitivity to DNA damage in isogenic teratocarcinoma cells. AB - Teratocarcinomas are tumors that arise from primordial germ cells and are readily curable with DNA-damaging chemotherapeutic drugs. Teratocarcinoma cells ex vivo in tissue culture are also relatively chemosensitive and undergo apoptotic death in response to DNA damage. We have previously hypothesized that the observed sensitivity of this tumor type to DNA damage is related to high basal expression of wild-type p53 protein. We have now addressed this issue by characterizing the DNA damage response of isogenic teratocarcinoma cells that differ only in their level of expression of wild-type p53 protein. We find a clear p53 dose-response relationship in these cells for rapid apoptosis following DNA damage that correlates with diminished colony formation in clonogenic survival assays. These results suggest that strategies to increase basal wild-type p53 protein expression prior to treatment with DNA-damaging drugs may improve curability in other tumor types. PMID- 11420713 TI - Chromatin remodeling: why it is important in cancer. AB - A typical human cell expresses only a few thousand of the more than 30 000 genes contained within our chromosomes. The chromosomal infrastructure is essential for gene control, determining both active and repressed states. It is important not only to turn the right genes on but also to turn the right genes off. Histones and chromatin components have key roles in this decision making process. Mistakes have severe consequences. If as few as three inappropriate genes are turned off, a normal cell can be converted into a cancer cell. This epigenetic silencing of genes underlies a new approach to cancer therapy. Advances in the biochemistry and genetics of chromatin remodeling reveal that gene inactivation depends on the recruitment of enzymes that control the display of DNA within the chromosome. Mistargeting of these enzymes leads to tumorigenesis, but inhibition of their activity presents a novel approach to therapy. PMID- 11420714 TI - Chromatin remodeling and transcriptional activation: the cast (in order of appearance). AB - The number of chromatin modifying and remodeling complexes implicated in genome control is growing faster than our understanding of the functional roles they play. We discuss recent in vitro experiments with biochemically defined chromatin templates that illuminate new aspects of action by histone acetyltransferases and ATP-dependent chromatin remodeling engines in facilitating transcription. We review a number of studies that present an 'ordered recruitment' view of transcriptional activation, according to which various complexes enter and exit their target promoter in a set sequence, and at specific times, such that action by one complex sets the stage for the arrival of the next one. A consensus emerging from all these experiments is that the joint action by several types of chromatin remodeling machines can lead to a more profound alteration of the infrastructure of chromatin over a target promoter than could be obtained by these enzymes acting independently. In addition, it appears that in specific cases one type of chromatin structure alteration (e.g., histone hyperacetylation) is contingent upon prior alterations of a different sort (i.e., ATP-dependent remodeling of histone-DNA contacts). The striking differences between the precise sequence of action by various cofactors observed in these studies may be - at least in part - due to differences between the specific promoters studied, and distinct requirements exhibited by specific loci for chromatin remodeling based on their pre-existing nucleoprotein architecture. PMID- 11420715 TI - An embarrassment of niches: the many covalent modifications of histones in transcriptional regulation. PMID- 11420716 TI - Role of protein methylation in chromatin remodeling and transcriptional regulation. AB - Recent findings suggest that lysine and arginine-specific methylation of histones may cooperate with other types of post-translational histone modification to regulate chromatin structure and gene transcription. Proteins that methylate histones on arginine residues can collaborate with other coactivators to enhance the activity of specific transcriptional activators such as nuclear receptors. Lysine methylation of histones is associated with transcriptionally active nuclei, regulates other types of histone modifications, and is necessary for proper mitotic cell divisions. The fact that some transcription factors and proteins involved in RNA processing can also be methylated suggests that protein methylation may also contribute in other ways to regulation of transcription and post-transcriptional steps in gene regulation. In future work, it will be important to develop methods for evaluating the precise roles of protein methylation in the regulation of native genes in physiological settings, e.g. by using chromatin immunoprecipitation assays, differentiating cell culture systems, and genetically altered cells and animals. It will also be important to isolate additional protein methyltransferases by molecular cloning and to characterize new methyltransferase substrates, the regulation of methyltransferase activities, and the roles of new methyltransferases and substrates. PMID- 11420717 TI - Histone H3 phosphorylation and cell division. AB - Histone H3 is specifically phosphorylated during both mitosis and meiosis in patterns that are specifically coordinated in both space and time. Histone H3 phosphorylation may initiate at different phases of the cell division in different organisms, but metaphase chromosomes are always found to be heavily phosphorylated. Upon exit of mitosis/meiosis a global dephosphorylation of H3 takes place. Potential candidates for H3 kinases are described and their hypothetical mechanism of action on highly condensed chromatin templates is discussed. In addition, a novel hypothesis for the role of histone H3 phosphorylation during cell division is proposed. This hypothesis, termed the 'ready production label' model, explains the results in the literature and suggests that phosphorylation of histone H3 is a part of a complex signaling mechanism. PMID- 11420718 TI - In vivo analysis of the model tyrosine aminotransferase gene reveals multiple sequential steps in glucocorticoid receptor action. AB - We are studying the mechanisms of transcriptional activation by nuclear receptors and we focus our studies on the glucocorticoid regulation of the model tyrosine aminotransferase gene. Rather than using in vitro biochemical approaches, we determine the actual events occurring in the cells. Our experimental approaches include genomic footprinting, chromatin immunoprecipitation, in situ hybridization and transgenic mice. Our results show that the glucocorticoid receptor uses a dynamic multistep mechanism to recruit successively accessory DNA binding proteins that assist in the activation process. Chromatin is first remodelled, DNA is then demethylated, and the synthesis of an accessory factor is induced. Efficient transcription induction is finally achieved upon the formation of a 'stable' multiprotein complex interacting with the regulatory element. We discuss: the relative contribution of histone acetyltransferases and ATP dependent remodelling machines to the chromatin remodelling event; the nature of the remodelled state; the contribution of regulated DNA demethylation to gene memory during development; the mechanisms of regulated DNA demethylation; the dynamics of protein recruitment at regulatory elements; the control of the frequency of transcription pulses and the control levels of the cell-type specificity of the glucocorticoid response. PMID- 11420719 TI - Glucocorticoid receptor-mediated chromatin remodeling in vivo. AB - The compaction of DNA into chromatin provides an additional level of gene regulation in eukaryotes that may not be available to prokaryotes. When packaged as chromatin, most promoters are transcriptionally repressed, and transcription factors have reduced access to their binding sites. The glucocorticoid receptor (GR) is a ligand-activated transcription factor that regulates the activity of genes involved in many physiological processes. To regulate eukaryotic genes, the GR binds to target sites within promoter regions of genes assembled as chromatin. This interaction alters the nucleosomal architecture to allow binding of other transcription factors, and formation of the preinitiation complex. The mouse mammary tumor virus (MMTV) promoter has been used extensively as a model to explore the processes by which the GR remodels chromatin and activates transcription. Significant progress has been made in our understanding of the mechanisms used by the GR to modify chromatin structure, and the limits placed on the GR by post-translational modifications of histones. We will describe recent developments in the processes used by the GR to activate transcription in vivo via chromatin remodeling complexes, histone H1 phosphorylation, and recruitment of diverse coactivators. PMID- 11420720 TI - Nuclear receptors coordinate the activities of chromatin remodeling complexes and coactivators to facilitate initiation of transcription. AB - Recent advances in the field of in vitro chromatin assembly have led to in vitro transcription systems which reproduce in the test tube, in vivo characteristics of ligand-dependent transcriptional activation by nuclear receptors. Dissection of these systems has begun to provide us with information concerning the underlying molecular mechanisms. Through recruitment of coactivator proteins, nuclear receptors act first to remodel chromatin within the promoter region and then to recruit the transcriptional machinery to the promoter region in order to initiate transcription. Here we present a possible sequential mechanism for ligand-dependent transcriptional activation by nuclear receptors and discuss the in vitro and in vivo data that support this model. PMID- 11420721 TI - Chromatin silencing and activation by Polycomb and trithorax group proteins. AB - The Polycomb group (PcG) of repressors and the trithorax group (trxG) of activators maintain the correct expression of several key developmental regulators, including the homeotic genes. PcG and trxG proteins function in distinct multiprotein complexes that are believed to control transcription by changing the structure of chromatin, organizing it into either a 'closed' or an 'open' conformation. The hallmark of gene regulation by PcG/trxG proteins is that it can lead to a mitotically stable pattern of gene expression, often referred to as epigenetic regulation. Although much remains to be learned, recent studies have provided insights into how this epigenetic switch is set, how PcG/trxG proteins might be linked to cis-acting DNA elements and what potential mechanisms underlie stable inheritance of gene expression status over multiple cell divisions. Finally, the study of the evolutionarily conserved PcG/trxG factors has recently gained additional urgency with the realization that they play a pertinent role in certain human cancers. PMID- 11420722 TI - When the SWI/SNF complex remodels...the cell cycle. AB - Mammalian cells contain several chromatin-remodeling complexes associated with the Brm and Brg1 helicase-like proteins. These complexes likely represent the functional homologs of the SWI/SNF and RSC complexes found in Saccharomyces cerevisiae. The mammalian chromatin-remodeling complexes are involved in both activation and repression of a variety of genes. Several lines of evidence also indicate that they play a specific role in the regulation of cell growth. Brm is down-regulated by ras signaling and its forced re-expression suppresses transformation by this oncogene. Besides, the Brg1 gene is silenced or mutated in several tumors cell lines and a Brg1-associated complex was recently found to co purify with BRCA1, involved in breast and ovarian cancers. Finally, the gene encoding SNF5/Ini1, a subunit common to all mammalian SWI/SNF complexes, is inactivated in rhabdoid sarcomas, a very aggressive form of pediatric cancer. The current review will address observations made upon inactivation of Brm, Brg1 and SNF5/Ini1 by homologous recombination in the mouse, as well as the possible implication of these factors in the regulation of the Retinoblastoma pRb-mediated repression of the transcription factor E2F. PMID- 11420723 TI - ATP-dependent chromatin remodeling factors: nucleosome shufflers with many missions. AB - This review addresses recent developments in the field of ATP-dependent chromatin remodeling factors. These factors use the energy of ATP hydrolysis to introduce superhelical torsion into DNA, which suggests a common mechanistic basis of action. Chromatin remodeling factors function both in transcriptional activation and repression, but they may have roles outside of transcriptional regulation such as DNA repair. A study of the nucleosome dependent ATPase ISWI in yeast illustrates the involvement of ATP-dependent chromatin remodeling in transcriptional repression by setting up inaccessible chromatin structures at promoters. However, factors such as ISWI are also involved in the restructuring of large chromatin domains and even whole chromosomes. Transcriptional regulation by ATP-dependent chromatin remodeling factors occurs in concert with histone modifying enzymes such as histone acetyltransferases and histone deacetylases: In yeast, SWI/SNF targeting is a requirement for histone acetyltransferases activity at promoters that are active at late stages of mitosis, when the chromatin is still condensed. This demonstrates that ATP-dependent remodeling factors facilitate covalent histone modifications. However, they are also regulated by histone modifications and in some circumstances they function in parallel with histone modifications towards the same goal. PMID- 11420724 TI - Chromatin remodelling and DNA replication: from nucleosomes to loop domains. AB - Organization of DNA into chromatin is likely to participate in the control of the timing and selection of DNA replication origins. Reorganization of the chromatin is carried out by chromatin remodelling machines, which may affect the choice of replication origins and efficiency of replication. Replication itself causes a profound rearrangement in the chromatin structure, from nucleosomes to DNA loop domains, allowing to retain or switch an epigenetic state. The present review considers the effects of chromatin remodelling on replication and vice versa. PMID- 11420725 TI - Chromatin alteration, transcription and replication: What's the opening line to the story? AB - Polymerase accessibility to chromatin is a limiting step in both RNA and DNA synthesis. Unwinding DNA and nucleosomes during polymerase complex binding and processing likely requires priming by chromatin restructuring. The initiating step in these processes remains an area of speculation. This review focuses on the physical handling of chromatin during transcription and replication, the fate of nucleosomes assembled on DNA during unwinding and processing the chromatin substrate, and how these alterations in chromatin structure may affect gene expression. Transcription or replication may alter chromatin structure during synthesis, enabling regulatory factor binding and, potentially, future rounds of transcription. As chromatin remodeling and transcription factor binding augment transcription and replication, and are themselves increased by these processes, a temporal model of structural alterations and gene activation is built that may be more circular than linear. PMID- 11420726 TI - Avian erythroleukemia: a model for corepressor function in cancer. AB - Transcriptional regulation at the level of chromatin plays crucial roles during eukaryotic development and differentiation. A plethora of studies revealed that the acetylation status of histones is controlled by multi-protein complexes containing (de)acetylase activities. In the current model, histone deacetylases and acetyltransferases are recruited to chromatin by DNA-bound repressors and activators, respectively. Shifting the balance between deacetylation, i.e. repressive chromatin and acetylation, i.e. active chromatin can lead to aberrant gene transcription and cancer. In human acute promyelocytic leukemia (APL) and avian erythroleukemia (AEL), chromosomal translocations and/or mutations in nuclear hormone receptors, RARalpha [NR1B1] and TRalpha [NR1A1], yielded oncoproteins that deregulate transcription and alter chromatin structure. The oncogenic receptors are locked in their 'off' mode thereby constitutively repressing transcription of genes that are critical for differentiation of hematopoietic cells. AEL involves an oncogenic version of the chicken TRalpha, v ErbA. Apart from repression by v-ErbA via recruitment of corepressor complexes, other repressors and corepressors appear to be involved in repression of v-ErbA target genes, such as carbonic anhydrase II (CAII). Reactivation of repressed genes in APL and AEL by chromatin modifying agents such as inhibitors of histone deacetylase or of methylation provides new therapeutic strategies in the treatment of acute myeloid leukemia. PMID- 11420727 TI - Histone deacetylases: a common molecular target for differentiation treatment of acute myeloid leukemias? AB - Recent discoveries have identified key molecular events in the pathogenesis of acute promyelocytic leukemia (APL), caused by chromosomal rearrangements of the transcription factor RAR (resulting in a fusion protein with the product of other cellular genes, such as PML). Oligomerization of RAR, through a self-association domain present in PML, imposes an altered interaction with transcriptional co regulators (NCoR/SMRT). NCoR/SMRT are responsible for recruitment of histone deacetylases (HDACs), which is required for transcriptional repression of PML-RAR target genes, and for the transforming potential of the fusion protein. Oligomerization and altered recruitment of HDACs are also responsible for transformation by the fusion protein AML1-ETO, extending these mechanisms to other forms of acute myeloid leukemias (AMLs) and suggesting that HDAC is a common target for myeloid leukemias. Strikingly, AML1-ETO expression blocks retinoic acid (RA) signaling in hematopoietic cells, suggesting that interference with the RA pathway (genetically altered in APL) by HDAC recruitment may be a common theme in AMLs. Treatment of APLs with RA, and of other AMLs with RA plus HDAC inhibitors (HDACi), results in myeloid differentiation. Thus, activation of the RA signaling pathway and inhibition of HDAC activity might represent a general strategy for the differentiation treatment of myeloid leukemias. PMID- 11420728 TI - Transcription therapy for cancer. AB - In the post genome era it will soon be possible to associate a specific tumor type with a specific gene expression profile and to define each molecular lesion characteristic of any given cancer. It is intuitive that a successful therapeutic strategy for cancer should aim at blocking the aberrant biochemical activity triggered by the oncogene or the lack of tumor suppressor gene activity that ultimately leads to full-blown neoplastic transformation. However, an attractive alternative approach entails the blockade of the transcriptional consequences of such oncogenic activities irrespective of their original biochemical nature, thus antagonizing the key transcriptional events underlying cancer pathogenesis in any specific neoplastic cellular population. This approach is now rendered possible by major advances along several lines of investigation: (i) the possibility of analysing gene expression through high throughput methods; (ii) a more detailed knowledge of the regulatory regions and of the transcription factors that control gene expression also facilitated in the future by a comprehensive whole genome comparative analysis of these regulatory sequences; (iii) the ability of modulating gene expression at the single gene level through various approaches both pharmacological and biochemical; (iv) the opportunity of directly antagonizing the aberrant activities of oncogenic transcription factors through a detailed knowledge of their abnormal transcriptional function; (v) the possibility of validating, in vivo, in animal models the relevance for neoplastic transformation of specific transcriptional events as well as of testing the efficacy of 'transcription therapy' in faithful animal models of human cancer. Here, we will review the facts, the existing applications and the hypothesis underlying such therapeutic modality for cancer therapy. PMID- 11420729 TI - The Rb/chromatin connection and epigenetic control: opinion. AB - The balance between cell differentiation and proliferation is regulated at the transcriptional level. In the cell cycle, the transition from G1 to S phase (G1/S transition) is of paramount importance in this regard. Indeed, it is only before this point that cells can be oriented toward the differentiation pathway: beyond, cells progress into the cycle in an autonomous manner. The G1/S transition is orchestrated by the transcription factor E2F. E2F controls the expression of a group of checkpoint genes whose products are required either for the G1-to-S transition itself or for DNA replication (e.g. DNA polymerase alpha). E2F activity is repressed in growth-arrested cells and in early G1, and is activated at mid-to-late G1. E2F is controlled by the retinoblastoma tumor suppressor protein Rb. Rb represses E2F mainly by recruiting chromatin remodeling factors (histone deacetylases and SWI/SNF complexes), the DNA methyltransferase DNMT1, and a histone methyltransferase. This review will focus on the molecular mechanisms of E2F repression by Rb during the cell cycle and during cell-cycle exit by differentiating cells. A model in which Rb irreversibly represses E2F regulated genes in differentiated cells by an epigenetic mechanism linked to heterochromatin, and involving histone H3 and promoter DNA methylation, is discussed. PMID- 11420730 TI - Rb-mediated chromatin structure regulation and transcriptional repression. PMID- 11420731 TI - DNA methylation, methyltransferases, and cancer. AB - The field of epigenetics has recently moved to the forefront of studies relating to diverse processes such as transcriptional regulation, chromatin structure, genome integrity, and tumorigenesis. Recent work has revealed how DNA methylation and chromatin structure are linked at the molecular level and how methylation anomalies play a direct causal role in tumorigenesis and genetic disease. Much new information has also come to light regarding the cellular methylation machinery, known as the DNA methyltransferases, in terms of their roles in mammalian development and the types of proteins they are known to interact with. This information has forced a new view for the role of DNA methyltransferases. Rather than enzymes that act in isolation to copy methylation patterns after replication, the types of interactions discovered thus far indicate that DNA methyltransferases may be components of larger complexes actively involved in transcriptional control and chromatin structure modulation. These new findings will likely enhance our understanding of the myriad roles of DNA methylation in disease as well as point the way to novel therapies to prevent or repair these defects. PMID- 11420732 TI - DNA methylation, chromatin inheritance, and cancer. AB - Cancer is a process driven by the accumulation of abnormalities in gene function. While many of these changes are genetic, epigenetically mediated changes in gene expression are being increasingly appreciated. This latter process emphasizes the need to understand two key components of heritable, but reversible, modulation of gene promoter function that are closely tied to one another - formation of chromatin which modulates transcription and establishing patterns of DNA methylation. The link lies first in the recruitment to methylated cytosines of a family of methyl-CpG binding domain proteins (MBDs), which are direct transcriptional repressors and can complex with transcriptional corepressors including histone deacetylases (HDACs). Additionally, the proteins that catalyze DNA methylation, DNA methyltransferases (DNMTs), also directly repress transcription and associate with HDACs. Regulation of these above chromatin-DNA methylation interactions as a function of DNA replication timing is emerging as a key event in the inheritance of transcriptionally repressed domains of the genome. Importantly, synergy between HDAC activity and DNA methylation is operative for a key epigenetic abnormality in cancer cells, transcriptional silencing of tumor suppressor genes. This change has now been recognized for genes that are essential for normal regulation of virtually every major cell function including cell growth, differentiation, apoptosis, DNA repair, and cell cell, cell-substratum interaction. Understanding the molecular determinants of both normal and abnormal patterns of chromatin formation and DNA methylation thus holds great promise for our understanding of cancer and for means to better diagnose, prevent, and treat this disease. PMID- 11420733 TI - Methyl CpG binding proteins: coupling chromatin architecture to gene regulation. AB - A correlation between DNA methylation and transcriptional silencing has existed for many years. Recently, substantial progress has been reported in the search for proteins that interpret the regulatory information inherent in DNA methylation and translate this information into functional states, resulting in the identification of a family of highly conserved proteins, the MBD family. Direct connections between these proteins and histone modification enzymes have emerged as a common theme, implying that DNA methylation exerts its effects primarily through repressive chromatin architecture. Recent structural determinations of the DNA binding domain of two MBD family members, MeCP2 and MBD1, provide a framework to model the interactions of this family with DNA. Comparative sequence analysis and experimental DNA binding data can be interpreted using this structural framework allowing one to contrast the members of the MBD family with each other and to predict the properties of new family members. The identification of mutations in MeCP2, the founding member of this family, as causal for the neurological developmental disorder Rett Syndrome provides additional information regarding amino acid residues crucial to the functions of this interesting protein family. PMID- 11420734 TI - Bowel dysfunction following spinal cord injury. AB - STUDY DESIGN: Review. OBJECTIVES: To outline the present knowledge of bowel dysfunction following spinal injury, and look at future directions of management and research. SETTING: Spinal Unit and Colorectal Unit, Christchurch, New Zealand. METHODOLOGY: Review. RESULTS: The underlying physiology of colorectal motility and defecation is reviewed, and consequences of spinal cord injury on defecation are reported. A discussion of present management techniques is undertaken and new directions in management and research are suggested. CONCLUSION: There is need for more intervention in regard to bowel function that could improve quality of life, but there is also a need for more research in this area. PMID- 11420735 TI - A comparison of bowel care patterns in patients with spinal cord injury: upper motor neuron bowel vs lower motor neuron bowel. AB - STUDY DESIGN: A face-to-face interview survey. OBJECTIVE: To compare bowel care patterns in spinal cord injury (SCI) patients based on type of neurogenic bowel. SETTING: Department of Physical Medicine and Rehabilitation of a tertiary university hospital in Suwon, Korea. METHODS: Among chronic SCI patients, 22 patients with upper motor neuron bowel (UMNB) and 20 patients with lower motor neuron bowel (LMNB) participated in an interview survey for the evaluation of bowel care patterns. RESULTS: The patients with LMNB demonstrated increased frequency of defecation, increased frequency of fecal incontinence, increased use of oral medications for bowel care, increased required time for defecation and more diet modification than those with UMNB (P < 0.05). However, there was no significant difference in the subjective difficulty of bowel care. Among several available bowel care methods, suppositories were used most frequently by the UMNB group, whereas the Valsalva maneuver was the most frequently used method by the LMNB group. CONCLUSIONS: Patients with LMNB tend to suffer more difficulties in management of their neurogenic bowel than those with UMNB. Therefore, more intensive and aggressive bowel care programs should be provided for SCI patients with LMNB. PMID- 11420736 TI - Osteoporosis and risk of fracture in men with spinal cord injury. AB - STUDY DESIGN: Cross-sectional study to evaluate bone mineral density (BMD) and fracture history after spinal cord injury (SCI). OBJECTIVES: To determine frequency of osteoporosis and fractures after SCI, correlate extent of bone loss with frequency of fractures after SCI, and determine fracture risk in SCI patients. SETTING: The Hines Veterans Affairs Hospital in Hines, Illinois, USA. METHODS: Femoral neck BMD was measured in 41 individuals with a history of traumatic or ischemic SCI using dual-energy X-ray absorptiometry (DEXA Lunar Whole Body Densitometer Model). RESULTS: Twenty-five patients (61%) met the World Health Organization (WHO) criteria for osteoporosis, eight (19.5%) were osteopenic, and eight (19.5%) were normal. Fracture after SCI had occurred in 14 patients (34%). There were significant differences between the femoral neck BMD and SCI duration in patients with a fracture history compared to those without. For patients in the same age group, each 0.1 gm/cm(2) and each unit of standard deviation (SD) (t-value) decrement of BMD at the femoral neck increased the risk of fracture 2.2 and 2.8 times, respectively. Considered simultaneously with age, duration of SCI, and level of SCI, BMD was the only significant predictor of the number of fractures. CONCLUSION: Osteoporosis and an increased frequency of fractures occur after SCI. Measurement of femoral neck BMD can be used to quantify fracture risk in SCI patients. PMID- 11420737 TI - Protective effect of glove on median nerve compression in the carpal tunnel. AB - OBJECTIVES: To determine the protective effect of gel padded glove on median nerve compression in the carpal tunnel. METHODS: Median nerve conduction parameters, skin temperature, laser Doppler flowmetry and pain modifications were measured during and after a 30-min carpal tunnel external compression protocol performed with and without glove in a random order on six healthy volunteers. RESULTS: Compression induced a rapidly reversible increase in sensory and motor distal latencies, a decrease in sensory amplitude, finger laser Doppler flowmetry and hand skin temperature supporting the hypothesis of a reversible conduction block of ischemic origin. There was no statistical difference between the tests (with or without glove) except for pain that was significantly reduced by glove protection. CONCLUSION: Gel padded glove does not seem to have a protective effect on the carpal tunnel syndrome induced by compression but provides significant comfort. PMID- 11420738 TI - Management of spinal brucellosis and outcome of rehabilitation. AB - STUDY DESIGN: Review of cases. OBJECTIVE: To review cases of brucellosis in order to clarify diagnostic guidelines, treatment regimes and prognosis. SETTING: University Hospital, Turkey. METHODS: Study of 11 patients (9 male, 2 female) with either brucellar spondylitis or epidural brucellar abscess. Diagnosis made on clinical presentation, laboratory findings, radiographic evidence and a minimum brucellar anti body of 1 : 160, a positive bacteriological culture and/or histological finding of inflammation of granulomatous tissue. All patients were treated with a combination of oral antibiotics. Surgery was performed in 8 patients. RESULTS: At least 6 months antibiotic therapy using Rifampicin and Doxycycline is recommended. Surgery is indicated in the presence of spinal instability, cord compression or radiculopathy. In our series eight patients required surgery. CONCLUSION: The patients complaining of back pain, particularly in endemic areas should be investigated as possible cases of brucellosis. PMID- 11420739 TI - Headaches in the whiplash syndrome. AB - STUDY DESIGN: A prospective clinical analysis of headaches in whiplash injury. OBJECTIVES: To provide a detailed clinical account of the nature, characteristics and natural history of headaches, and to examine their possible relation to the neck injury. SETTING: One medicolegal practice in UK. METHODS: This study consists of a prospective clinical analysis of headache symptoms in consecutive patients referred to the author for medicolegal assessment of whiplash injury, with no special reference to headaches. RESULTS: All 80 patients noted neck pain. Headaches were related as a consequence of the accident by 48 patients (60% of the total). The circumstances of injury did not differ from whiplash victims who did not have prominent headache. The headache onset was maximum in the first 24 h after injury. The failure to recall well documented pre-accident headaches in almost one fifth of patients is significant. The common types of headaches were non-specific, generalised, dull, aching pain (25 patients), a mixtures of aching and tightness, and tension type headache. Only three (6%) had migraine without aura. CONCLUSION: Post-whiplash headache is a genuine common but short lived affair, constant headaches disappearing within 3 weeks in 85% subjects. In the minority complaining of headache after that period there was no evidence of persisting physical injury, nor of inability to work or other disability caused by headaches. PMID- 11420740 TI - Pressure ulcers: an unusual complication of indwelling urethral catheter. AB - INTRODUCTION: Pressure ulcers are common among patients with spinal cord disorders (SCD) and occur due to unrelieved pressure on soft tissues. CASE REPORTS: Two ladies with paraplegia following acute transverse myelitis developed pressure ulcers over medial aspects of thighs due to indwelling urethral catheter. Absence of sensation, weakness of both legs and lack of knowledge about catheter care contributed to ulcer formation. CONCLUSION: Indwelling urethral catheter may unusually result in pressure ulcers over the thighs in patients with SCD. Among health professionals involved in the care of these subjects awareness is essential for preventing this complication. PMID- 11420741 TI - The occurrence of the ilio-lumbar syndrome in a spinal cord injury patient. AB - STUDY DESIGN: Case report of a 71-year-old man having sustained a spinal cord injury (SCI) for 26 years. He started to suffer from low back pain 21 years after his onset and this was diagnosed as ilio-lumbar syndrome (ILS). OBJECTIVES: To describe the problem of low back pain occurring after a change of wheelchair in a SCI, and to indicate the importance of the ilio-lumbar syndrome as a possible cause of low back complaint. OUTCOME: Clinical findings supported by echotomography confirmed the diagnosis of ILS in our case. CONCLUSION: ILS should be considered when some changes and deterioration of the status of SCI occur due to low back complaint. PMID- 11420742 TI - Complication of penile sheath drainage in a spinal cord injury patient: calculus impacting in the urethra proximal to the rim of a condom. PMID- 11420744 TI - Loss of expression of human spectrin src homology domain binding protein 1 is associated with 10p loss in human prostatic adenocarcinoma. AB - The gene encoding human spectrin Src homology domain binding protein 1, or Hssh3bp1, which is a marker of macropinocytic vesicles and a potential regulator of macropinocytosis, co-localizes to a YAC containing chromosome 10p sequences at loci D10S89 and D10S111 that are frequently deleted in prostate tumors. Expression of Hssh3bp1 was evaluated at the protein level in 17 paired normal and malignant prostate tumor samples using the monoclonal antibody 2G8 to Hssh3bp1. These experiments demonstrated that 4/6 tumors (67%) with 10p deletion failed to express Hssh3bp1 protein compared to 5/11 (46%) tumors with intact 10p. Thus, loss of Hssh3bp1 expression is concordant with allelic loss of adjacent 10p sequences in human prostate tumors. In addition, two prostate tumor cell lines contain an exon skipping mutation in the Hssh3bp1 gene that leads to the abnormal splicing of the mRNA and loss of a portion of Abl tyrosine kinase SH3 domain binding site in the protein. These data are consistent with a role for Hssh3bp1 as a candidate tumor suppressor gene inactivated during prostate tumorigenesis. PMID- 11420745 TI - Detection of MYCN gene amplification in neuroblastoma by fluorescence in situ hybridization: a pediatric oncology group study. AB - To assess the utility of fluorescence in situ hybridization (FISH) for analysis of MYCN gene amplification in neuroblastoma, we compared this assay with Southern blot analysis using tumor specimens collected from 232 patients with presenting characteristics typical of this disease. The FISH technique identified MYCN amplification in 47 cases, compared with 39 by Southern blotting, thus increasing the total number of positive cases by 21%. The major cause of discordancy was a low fraction of tumor cells (< or =30% replacement) in clinical specimens, which prevented an accurate estimate of MYCN copy number by Southern blotting. With FISH, by contrast, it was possible to analyze multiple interphase nuclei of tumor cells, regardless of the proportion of normal peripheral blood, bone marrow, or stromal cells in clinical samples. Thus, FISH could be performed accurately with very small numbers of tumor cells from touch preparations of needle biopsies. Moreover, this procedure allowed us to discern the heterogeneous pattern of MYCN amplification that is characteristic of neuroblastoma. We conclude that FISH improves the detection of MYCN gene amplification in childhood neuroblastomas in a clinical setting, thus facilitating therapeutic decisions based on the presence or absence of this prognostically important biologic marker. PMID- 11420746 TI - An N-terminal region of mot-2 binds to p53 in vitro. AB - The mouse mot-2 protein was earlier shown to bind to the tumor suppressor protein, p53. The mot-2 binding site of p53 was mapped to C-terminal amino acid residues 312-352, which includes the cytoplasmic sequestration domain. In the present study, we have found that both mot-1 and mot-2 bind to p53 in vitro. By using His-tagged deletion mutant proteins, the p53-binding domain of mot-2 was mapped to its N-terminal amino acid residues 253-282, which are identical in mot 1 and mot-2 proteins. Some peptides containing the p53-binding region of mot-2 were able to compete with the full-length protein for p53 binding. The data provided rationale for in vitro binding of mot-1 and mot-2 proteins to p53 and supported the conclusion that inability of mot-1 protein to bind p53 in vivo depends on secondary structure or its binding to other cellular factors. Most interestingly, the p53-binding region of mot-2 was common to its MKT-077, a cationic dye that exhibits antitumor activity, binding region. Therefore it is most likely that MKT-077-induced nuclear translocation and restoration of wild type p53 function in transformed cells takes place by a competitional mechanism. PMID- 11420747 TI - Tumor cells express FcgammaRI which contributes to tumor cell growth and a metastatic phenotype. AB - High levels of circulating immune complexes containing tumor-associated antigens are associated with a poor prognosis for individuals with cancer. The ability of B cells, previously exposed to tumor-associated antigens, to promote both in vitro and in vivo tumor growth formed the rationale to evaluate the mechanism by which immune complexes may promote tumor growth. In elucidating this mechanism, FcgammaRI expression by tumor cells was characterized by flow cytometry, polymerase chain reaction, and sequence analysis. Immune complexes containing shed tumor antigen and anti-shed tumor antigen Ab cross-linked FcgammaRI expressing tumor cells, which resulted in an induction of tumor cell proliferation and of shed tumor antigen production. Use of selective tyrosine kinase inhibitors demonstrated that tumor cell proliferation induced by immune complex cross-linking of FcgammaRI is dependent on the tyrosine kinase signal transduction pathway. A selective inhibitor of phosphatidylinositol-3 kinase also inhibited this induction of tumor cell proliferation. These findings support a role for immune complexes and FcgammaRI expression by tumor cells in augmentation of tumor growth and a metastatic phenotype. PMID- 11420748 TI - Resistance of muscle to tumor metastases: a role for a3 adenosine receptor agonists. AB - Tumor metastases are extremely rare in striated muscles. Lately, we have found that muscle cell conditioned medium (MCM) inhibits the proliferation of various tumor cells while maintaining the growth of normal murine bone marrow cells. This dual activity was confirmed in vivo when the MCM was administered orally, i.e., it inhibited the development of tumor growth in mice and prevented the myelotoxic effects of chemotherapy. Adenosine was found to be one of the active components of MCM, inhibiting tumor cell growth while maintaining bone marrow cell proliferation in vitro. Adenosine is known to act as an important regulatory molecule through its binding to specific G-protein-associated A1, A(2a), A(2b) and A3 cell surface receptors. In distinction from MCM, adenosine did not suppress tumor development in mice and was not active as a chemoprotective agent when administered orally or intravenously. Thus, the in vivo activity of MCM could not be attributed to adenosine. In this study, MCM from which adenosine was enzymatically removed still retained its dual activity that was also found to be mediated through the A3 adenosine receptor (A3AR). This result led to the conclusion that natural agonists to A3AR were responsible for the activity of MCM. We further tested synthetic agonist to the A3AR and demonstrated that it possessed the same in vitro and in vivo activity profile as MCM. Taken together, muscle cells, in addition to adenosine, secrete natural agonists to A3AR. These agonists are stable nondegradable molecules and may contribute to the systemic anticancer and chemoprotective activity exerted by MCM. This group of molecules may account for the rarity of tumor metastases in muscle. PMID- 11420749 TI - Detection of abundantly transcribed genes and gene translocation in human immunodeficiency virus-associated non-Hodgkin's lymphoma. AB - Several novel, differentially transcribed genes were identified in one centroblastic and one immunoblastic HIV-associated B-cell non-Hodgkin's lymphoma (B-NHL) by subtractive cloning. In both lymphomas, we detected an upregulated transcription of several mitochondrial genes. In the centroblastic B-NHL, we found a high level transcription of nuclear genes including the interferon inducible gene (INF-ind), the immunoglobulin light chain gene (IgL), the set oncogene, and several unknown genes. The data obtained on upregulated expression of the genes in human B-NHL of HIV-infected patients considerably overlap with those obtained earlier for the B-NHL of simian immunodeficiency virus-infected monkeys. In the centroblastic lymphoma, one transcript revealed a fusion of the 3'-untranslated region of the set gene and the C-terminal region of the IgL gene. This chimeric sequence was confirmed by a site-directed polymerase chain reaction performed with total cDNA and genomic DNA. The expected amplification product was obtained in both cases pointing to a genomic rearrangement. The IgL-set fusion sequence was not found in cDNA preparations and genomic DNA of the immunoblastic HIV-associated B-NHL. Further studies are necessary to determine whether these genes contribute to lymphoma development or can be used as therapeutic targets. PMID- 11420750 TI - Vascular differences detected by MRI for metastatic versus nonmetastatic breast and prostate cancer xenografts. AB - Several studies have linked vascular density, identified in histologic sections, to "metastatic risk." Functional information of the vasculature, not readily available from histologic sections, can be obtained with contrast-enhanced MRI to exploit for therapy or metastasis prevention. Our aims were to determine if human breast and prostate cancer xenografts preselected for differences in invasive and metastatic characteristics established correspondingly different vascular volume and permeability, quantified here with noninvasive MRI of the intravascular contrast agent albumin-GdDTPA. Tumor vascular volume and permeability of human breast and prostate cancer xenografts were characterized using MRI. Parallel studies confirmed the invasive behavior of these cell lines. Vascular endothelial growth factor (VEGF) expression in the cell lines was measured using ELISA and Western blots. Metastasis to the lungs was evaluated with spontaneous as well as experimental assay. Metastatic tumors formed vasculature with significantly higher permeability or vascular volume (P<.05, two-sided unpaired t test). The permeability profile matched VEGF expression. Within tumors, regions of high vascular volume usually exhibited low permeability whereas regions of low vascular volume exhibited high permeability. We observed that although invasion was necessary, without adequate vascularization it was not sufficient for metastasis to occur. PMID- 11420752 TI - DNA fragmentation factor 45 (DFF45) gene at 1p36.2 is homozygously deleted and encodes variant transcripts in neuroblastoma cell line. AB - Recently, loss of heterozygosity (LOH) studies suggest that more than two tumor suppressor genes lie on the short arm of chromosome 1 (1p) in neuroblastoma (NB). To identify candidate tumor suppressor genes in NB, we searched for homozygous deletions in 20 NB cell lines using a high-density STS map spanning chromosome 1p36, a common LOH region in NB. We found that the 45-kDa subunit of the DNA fragmentation factor (DFF45) gene was homozygously deleted in an NB cell line, NB 1. DFF45 is the chaperon of DFF40, and both molecules are necessary for caspase 3 to induce apoptosis. DFF35, a splicing variant of DFF45, is an inhibitor of DFF40. We examined 20 NB cell lines for expression and mutation of DFF45 gene by reverse transcription (RT)-polymerase chain reaction (PCR) and RT-PCR-single strand conformation polymorphism. Some novel variant transcripts of the DFF45 gene were found in NB cell lines, but not in normal adrenal gland and peripheral blood. These variants may not serve as chaperons of DFF40, but as inhibitors like DFF35, thus disrupting the balance between DFF45 and DFF40. No mutations of the DFF45 gene were found in any NB cell line, suggesting that the DFF45 is not a tumor suppressor gene for NB. However, homozygous deletion of the DFF45 gene in the NB-1 cell line may imply the presence of unknown tumor suppressor genes in this region. PMID- 11420751 TI - Suppression of angiogenesis and therapy of human colon cancer liver metastasis by systemic administration of interferon-alpha. AB - The purpose of this study was to determine whether systemic administration of interferon-alpha (IFN-alpha) can inhibit liver metastasis produced in nude mice by human colon cancer cells. KM12L4 (IFN-alpha-sensitive) or KM12L4 IFN(R) (IFN alpha-resistant) cells were injected into the spleen of nude mice. Seven days later, the mice were treated with subcutaneous (s.c.) injections of IFN-alpha (70,000 units/week) at different dosing schedules (1, 2, or 7 times/week). Significant inhibition of tumor growth, vascularization and expression of basic fibroblast growth factor (bFGF) or matrix metalloproteinase-9 (MMP-9) mRNA and protein occurred in mice given daily injections of IFN-alpha. Kinetic analysis of therapy showed that daily s.c. administrations of 10,000 units of IFN-alpha induced apoptosis in liver metastasis-associated endothelial cells, followed by inhibition of tumor cell division and apoptosis of tumor cells. These data suggest that the antiangiogenic activity of IFN-alpha-2a depends on frequent administration of the optimal biologic dose. PMID- 11420754 TI - Tracking issue attention: specifying the dynamics of the public agenda. AB - While interest in issues shifts daily, tracking these changes is currently done by multiple surveys, which are conducted months apart. In this article, we adapt political tracking poll methods to model the functional form of issue attention changes through three phases, including equilibrium, developing interest, and decline. We find that opinions do follow discernible cycles. Further, we test and find strong support for the "boredom" explanation that reactions to external events that are brought to the attention of the public through the media are much greater during the developing interest phase than the saturation phase. The evidence gathered from the newly adapted method has implications for modeling the issue-attention cycle and testing the effects of media coverage and external events on public interest in issues. PMID- 11420755 TI - Who Votes by Mail?: A Dynamic Model of the Individual-Level Consequences of Voting-by-Mail Systems. AB - Election administrators and public officials often consider changes in electoral laws, hoping that these changes will increase voter turnout and make the electorate more reflective of the voting-age population. The most recent of these innovations is voting-by-mail (VBM), a procedure by which ballots are sent to an address for every registered voter. Over the last 2 decades, VBM has spread across the United States, unaccompanied by much empirical evaluation of its impact on either voter turnout or the stratification of the electorate. In this study, we fill this gap in our knowledge by assessing the impact of VBM in one state, Oregon. We carry out this assessment at the individual level, using data over a range of elections. We argue that VBM does increase voter turnout in the long run, primarily by making it easier for current voters to continue to participate, rather than by mobilizing nonvoters into the electorate. These effects, however, are not uniform across all groups in the electorate. Although VBM in Oregon does not exert any influence on the partisan composition of the electorate, VBM increases, rather than diminishes, the resource stratification of the electorate. Contrary to the expectations of many reformers, VBM advantages the resource-rich by keeping them in the electorate, and VBM does little to change the behavior of the resource-poor. In short, VBM increases turnout, but it does so without making the electorate more descriptively representative of the voting-age population. PMID- 11420756 TI - Economic class and popular support for franklin roosevelt in war and peace. AB - History has judged FDR as one of America's greatest leaders in part because he maintained the public's confidence in seeing the nation through the travails of the Depression and World War II. During this era, the two most widely employed explanatory variables in contemporary presidential popularity scholarship-the economy and war-assumed their most extreme values of the twentieth century. Hence, not only is understanding Roosevelt's public support historically important, but it represents a valuable case for filling in our understanding of the opinion dynamics of presidential support more generally. Yet, surprisingly, Roosevelt's approval ratings have attracted little systematic scrutiny. Compiling time-series data from 1937 to 1943, partially disaggregated by economic class, we investigate FDR's popular support among different classes during both national crises. We find that Roosevelt's peacetime support divided along class lines, while during the war class divisions blurred. Roosevelt's support was indeed conditioned by external events, refracted through the interests of different societal groups. We conclude that public support for modern presidents should be similarly studied as the sum of opinions among heterogeneous constituencies. PMID- 11420757 TI - Web survey design and administration. AB - Many claims are being made about the advantages of conducting surveys on the Web. However, there has been little research on the effects of format or design on the levels of unit and item response or on data quality. In a study conducted at the University of Michigan, a number of experiments were added to a survey of the student population to assess the impact of design features on resulting data quality. A sample of 1,602 students was sent an e-mail invitation to participate in a Web survey on attitudes toward affirmative action. Three experiments on design approaches were added to the survey application. One experiment varied whether respondents were reminded of their progress through the instrument. In a second experiment, one version presented several related items on one screen, while the other version presented one question per screen. In a third experiment, for one series of questions a random half of the sample clicked radio buttons to indicate their answers, while the other half entered a numeric response in a box. This article discusses the overall implementation and outcome of the survey, and it describes the results of the imbedded design experiments. PMID- 11420758 TI - Urban chinese perceptions of threats from the United States and Japan. PMID- 11420759 TI - The Polls-Trends: Attitudes about Agricultural Biotechnology and Genetically Modified Organisms. PMID- 11420760 TI - The fallacy of the mean. PMID- 11420761 TI - Hemodynamic profiles of advanced heart failure: association with clinical characteristics and long-term outcomes. AB - BACKGROUND: Classifying patients with advanced congestive heart failure (CHF) by baseline measures of congestion and perfusion has been used to estimate hemodynamic status and to select and titrate therapy. We describe clinical characteristics of 4 hemodynamic profiles-wet/cold, wet/warm, dry/cold, and dry/warm-in patients with advanced CHF and assess relations between symptoms, physical signs, and outcomes with each profile. METHODS AND RESULTS: We retrospectively assessed baseline symptoms, physical-examination variables, and 1 year outcomes of 440 patients in a randomized trial. With univariable and multivariable logistic regression, we examined relations of physical-examination variables to hemodynamic profiles. We also assessed the rates of death and death or readmission by profile. Severity of CHF symptoms did not predict the wet versus-dry profile or cold-versus-warm status, despite significant differences in hemodynamics among groups. Of the physical-examination variables, only a lower proportional pulse pressure was a significant multivariable predictor of the wet category. Among wet patients (n = 348), this same variable was the only significant multivariable predictor of the cold category. For dry patients (n = 92), the cold category was predicted in multivariable analysis by supine heart rate and hepatomegaly. Survival was similar among profiles: wet/cold, 54.2% (n = 91); wet/warm, 58.3% (n = 105); dry/cold, 78.9% (n = 15); and dry/warm, 67.1%, P =.13 (n = 49). Event-free survival also was similar among profiles: wet/cold, 22.0% (n = 37); wet/warm, 29.4% (n = 53); dry/cold, 42.1% (n = 8); and dry/warm, 31.5%, P =.44 (n = 23). CONCLUSIONS: The patient's history and physical examination alone may lead to inaccurate estimation of hemodynamic status and thus suboptimal management for patients with advanced CHF. PMID- 11420762 TI - A randomized multicenter study comparing the efficacy and safety of intravenous milrinone and intravenous nitroglycerin in patients with advanced heart failure. AB - A randomized, open-label, parallel-group design was used to determine the percentage of patients achieving improvements in predetermined baseline hemodynamic end points (>20% to 30% increase in cardiac index depending on baseline values and >25% decrease in pulmonary capillary wedge pressure), assessed at hour 0 (end of initial dose titration) and 1, 2, 4, 8, and 24 hours after the infusion of milrinone or nitroglycerin. In total, 125 patients (60 milrinone, 65 nitroglycerin) enrolled in this study, and 119 (58 milrinone, 61 nitroglycerin) were evaluable for the efficacy analysis. A significantly greater proportion of milrinone-treated patients reached (45% v 14%, P =.005) and maintained (24% v 6%, P =.026) hemodynamic goals than did nitroglycerin-treated patients; the time to achieve hemodynamic goals was significantly less in milrinone-treated patients (33 +/- 2 v 54 +/- 10 minutes, P <.001). Milrinone was also significantly more effective in decreasing systemic vascular resistance (P =.004), increasing stroke volume (P =.008), and improving global clinical status. Inodilator therapy with milrinone seems more efficacious in attaining sustained hemodynamic improvement than does pure intravenous vasodilator therapy with nitroglycerin in treating patients with decompensated heart failure. PMID- 11420763 TI - Activity of the neurohormonal system and its relationship to autonomic abnormalities in decompensated heart failure. AB - BACKGROUND: The syndrome of congestive heart failure (CHF) entails complex autonomic and hormonal responses. Profound abnormalities in autonomic function, characterized by sympathetic overactivity and parasympathetic withdrawal, exert direct deleterious effects on the heart and contribute to progressive circulatory failure. We investigated the relationship of heart rate variability (HRV) with levels of neurohormones in plasma. METHODS AND RESULTS: We studied 64 patients admitted to the hospital for treatment of decompensated CHF (mean age, 59 +/- 2 years; New York Heart Association class III [72%] and IV [28%]). Time- and frequency-domain HRV indices were obtained from 24-hour Holter recordings. Neurohormonal activation was assessed by measuring plasma renin activity and aldosterone and norepinephrine levels. In the time domain, norepinephrine correlated negatively with average NN interval (r = -.34; P =.007), SDNN (r = .35; P =.005), and SDANN (r = -.36; P =.004). In the frequency domain, norepinephrine was negatively associated with the total power (r = -.39; P =.001) and ultralow power (r = -.43; P =.0005). No correlation was found between indices indicative of parasympathetic modulation, except for a borderline correlation with the high-frequency power (r = -.25; P =.048). CONCLUSIONS: Reduced HRV may be associated with increased norepinephrine levels in patients with severe CHF. The ability of long-term HRV parameters to reflect in part the activation of diverse hormonal systems may explain their greater prognostic power for risk stratification in patients with CHF. PMID- 11420764 TI - beta-Adrenergic and endothelin receptor interaction in dilated human cardiomyopathic myocardium. AB - BACKGROUND: Although end-stage dilated cardiomyopathy (DCM) is characterized by defects in beta-adrenergic receptor (beta-AR) activity and increased endothelin-1 (ET-1), possible interactions between these 2 systems remain to be defined. Accordingly, the goal of this study was to determine the effects of ET receptor activation on beta-AR signaling through measurement of cyclic adenosine monophosphate (cAMP) in normal and DCM myocardium. METHODS AND RESULTS: Myocardial sarcolemmal preparations were prepared from normal human (n = 6), dilated cardiomyopathic (n = 10), and ischemic cardiomyopathic (ICM, n = 10) tissue. Basal cAMP production was measured in the presence of ET-1 alone (10(-6) to 0(-9) mol/L) as well as after (-)isoproterenol (10(-6) to 10(-2) mol/L) or forskolin (0.05 to 30.0 micromol/L) stimulation. beta-AR and ET receptor profiles were determined by radiolabeled ligand assays. ET-1 inhibited basal cAMP production in all preparations in a concentration-dependent manner. However, beta AR-stimulated cAMP production by either isoproterenol or forskolin was not significantly affected by ET-1. beta-AR receptor density was reduced, and a selective reduction of the ET(B) receptor occurred in both forms of DCM. CONCLUSIONS: Under basal conditions, ET receptor stimulation reduced cAMP levels, which may influence contractility, particularly with DCM. PMID- 11420765 TI - Mechanical alternans in patients with chronic heart failure. AB - BACKGROUND: Clinical implications of mechanical alternans in patients with chronic heart failure have remained uncertain. In this study, prevalence, characteristics, and prognostic implications of mechanical alternans were investigated. METHODS AND RESULTS: Consecutive 51 patients with dilated cardiomyopathy underwent diagnostic cardiac catheterization using a micromanometer-tipped catheter. Under basal conditions, 7 of 35 patients with sinus rhythm showed mechanical alternans. Physiologic tachycardia (110 bpm) induced mechanical alternans in another 15 patients with sinus rhythm and in another 10 of 16 patients with atrial fibrillation. Low doses of dobutamine also induced mechanical alternans in another 8 patients, but a high dose of dobutamine eliminated mechanical alternans. Consequently, 40 patients (78%) showed mechanical alternans. Mechanical alternans was always accompanied by alternating changes of positive dP/dt, a parameter of contractility during isovolumetric contraction time, but negative dP/dt was occasionally constant. Concordant mechanical alternans between both ventricles was more prevalent than discordant alternans. The left ventricular end-diastolic volume indices and end-systolic volume indices of patients with mechanical alternans were larger than those of patients without. The left ventricular ejection fraction of patients with alternans was significantly lower than that of patients without. CONCLUSIONS: Mechanical alternans was highly prevalent in patients with chronic heart failure. The origin of mechanical alternans seems to exist before or at the isovolumetric contraction time. PMID- 11420766 TI - Early manifestations of "sick euthyroid" syndrome in patients with compensated chronic heart failure. AB - BACKGROUND: A "sick euthyroid" syndrome occurs in patients with severe decompensated chronic heart failure (CHF) and other chronic illnesses and is related to adverse prognosis, but it has not been described in patients with compensated CHF. The aim of this study was to determine whether manifestations of the sick euthyroid syndrome occur in patients with compensated CHF caused by ischemic heart disease. METHODS AND RESULTS: Thyroid hormonal responses to thyrotropin-releasing hormone (TRH) stimulation were compared in 8 patients with New York Heart Association class I/II CHF considered secondary to ischemic heart disease and 7 control patients after serial 10-minute blood sampling over 3-hour periods. Secretory dynamics of TRH-induced thyroid-stimulating hormone (TSH) release were compared by using deconvolution analysis. Changes in serum thyroxine (T4), triiodothyronine (T3), reverse T3 (rT3), and rT3/T4 concentration ratios were compared. Patients with CHF had lower baseline serum T3 concentrations (P <.001), with lower maximum serum T(3) (P <.01) and higher maximum serum rT(3) (P <.05) concentrations after TRH stimulation but similar estimated TRH-induced TSH secretory burst amplitude, mass, and 3-hour production rates, compared with control patients. CONCLUSIONS: Patients with compensated CHF display the derangements in thyroid hormone metabolism of impaired peripheral conversion of T4 and T3 and increased production of rT(3) in the presence of normal dynamic function of the hypothalamic-pituitary-thyroid axis, which are consistent with early manifestations of a sick euthyroid state. PMID- 11420767 TI - A health perception score predicts cardiac events in patients with heart failure: results from the IMPRESS trial. AB - BACKGROUND: New York Heart Association (NYHA) class and treadmill exercise test variables are widely used for estimating prognosis and measuring the outcomes of treatment in patients with heart failure, but they do not take patients' perceptions into account. METHODS AND RESULTS: Five hundred forty-five patients enrolled in a multicenter 24-week comparison of the effects of omapatrilat and lisinopril on functional capacity in patients with heart failure reported a visual analog scale (VAS) score of their overall health perception at week 12 of the study. A total of 27 first events, defined as death or worsening heart failure (hospitalization, emergency room visit, or study discontinuation), occurred in the subsequent 12 weeks. The mean (+/-SD) health perception scores were 0.43 +/- 0.31 and 0.68 +/- 0.20 in patients with and without events, respectively (P =.0006). The risk ratio (RR) for an event associated with a decile change in the health perception score was 0.74 (95% confidence interval [CI], 0.61-0.88; P =.001). The RR was unaltered by adjustment for demographic variables, treadmill time, and NYHA functional class. Although the week 12 NYHA functional class was predictive of events (RR = 2.1; 95% CI, 1.2-4.6; P =.04), treadmill time was not (RR = 0.87; 95% CI, 0.73-1.03; P = 0.11). CONCLUSIONS: A patient-reported measure of perceived health predicts events in patients with heart failure. PMID- 11420768 TI - Comparison of amlodipine or nifedipine treatment with developing congestive heart failure: effects on myocyte contractility. AB - BACKGROUND: Past studies have suggested that amlodipine, a dihydropyridine L-type Ca(2+) channel antagonist, may exert useful effects in congestive heart failure (CHF). The present study examined the effects of amlodipine or nifedipine treatment in a model of developing CHF on left ventricular (LV) pump function and myocyte contractility. METHODS AND RESULTS: Pigs (25 kg) were randomly assigned to 1 of 4 groups: 1) pacing-induced CHF (rapid atrial pacing at 240 bpm) for 3 weeks (n = 9), 2) concomitant Ca(2+) channel blockade with amlodipine (1.5 mg/kg/day) and rapid pacing (n = 7), 3) concomitant Ca(2+) channel blockade with nifedipine (0.7 mg/kg twice daily) and rapid pacing (n = 7), and 4) sham controls (n = 7). LV fractional shortening fell with pacing CHF from baseline values (17% +/- 1% v 42% +/- 1%, P <.05). With rapid pacing and concomitant amlodipine treatment, LV fractional shortening increased from pacing CHF values (24% +/- 1%, P <.05) but was unchanged with concomitant nifedipine treatment (20% +/- 2%, P =.2). LV myocyte velocity of shortening, as measured by high speed videomicroscopy, was reduced with pacing CHF compared with controls (42 +/- 2 microm/s v 87 +/- 9 microm/s, P <.05), and increased from pacing CHF values with amlodipine or nifedipine treatment (62 +/- 8 microm/s, 64 +/- 4 microm/s, respectively; P <.05). Inotropic response to extracellular Ca(2+) (8 mmol/L) was reduced with pacing CHF (94 +/- 5 microm/s v 160 +/- 15 microm/s, P <.05) and increased from CHF values with amlodipine or nifedipine treatment (132 +/- 14 microm/s and 133 +/- 7 microm/s, respectively, P <.05) CONCLUSIONS: These results suggest that the primary mechanism for the effects of amlodipine on myocyte contractility in developing CHF is because of direct Ca(2+) channel blockade. PMID- 11420769 TI - UV light synergistically enhances the cardiotoxic effects of interleukin 1beta through peroxynitrite formation. AB - BACKGROUND: Proinflammatory cytokines play an important role in chronic cardiac diseases. METHODS AND RESULTS: Neonatal rat cardiomyocytes were exposed to interleukin (IL)-1beta (2 ng/mL) for 4 days. We assessed contractility through videomicroscopy and calcium transients with the Ca(2+)-sensitive dye fura-2. In IL-1beta-treated cells, the UV excitation (380 nm) necessary to induce dye fluorescence effected a rapid cessation of Ca(2+) transients and contraction, accompanied by calcium overload originating from an intracellular compartment. This occurred in the absence of fura-2 but required UV illumination. Incubation with 10 mmol/L N-acetylcysteine prevented this response, suggesting a free radical-mediated event. However, exposure to IL-1beta either increased or did not change the activity of the free radical scavengers superoxide dismutase, catalase, and glutathione peroxidase. In contrast, lipid peroxidation increased by 600% (P < or =.0001) in the IL-1beta plus UV-treated cells, an effect eliminated by L-NMMA. L-NMMA also completely abolished the UV-mediated cytotoxicity. We used immunohistochemistry to localize nitrotyrosine accumulation in the myocytes cotreated with IL-1beta and UV, an effect that was also blocked by L-NMMA. CONCLUSIONS: We hypothesize that the toxic radical peroxynitrite, arising from nitric oxide and superoxide anion, may be responsible for tetany and acute cardiomyocyte death. These results demonstrate the potential role of peroxynitrite in cardiotoxicity, which may be important in cardiac diseases associated with proinflammatory cytokines. PMID- 11420770 TI - Proposal for a new clinical end point to evaluate the efficacy of drugs and devices in the treatment of chronic heart failure. AB - Clinical trials designed to evaluate the effect of drugs and devices on the symptoms and clinical status in chronic heart failure have frequently produced conflicting, inconclusive, or misleading results. These difficulties can be explained by the fact that previous studies have relied on efficacy measures that have inherent limitations and have been analyzed using statistical approaches that ignored episodes of clinical deterioration. Recognition of these pitfalls has led to the development of a new clinical composite score, which combines changes in the New York Heart Association class and the global assessment together with the information provided from the occurrence of major clinical events. Use of this score would have correctly distinguished active therapy from placebo in earlier trials and thus would have avoided some of their misleading conclusions. The new clinical composite score has been prospectively incorporated into the design of studies evaluating the efficacy of endothelin antagonists, cytokine antagonists, vasopressin antagonists, and cardiac resynchronization in the treatment of chronic heart failure. In the trials that have been completed to date, the clinical composite score has been more sensitive than conventional approaches in discerning the presence or absence of a true treatment effect. PMID- 11420771 TI - B-type natriuretic peptide levels: a potential novel "white count" for congestive heart failure. AB - Finding a simple blood test to aid in the diagnosis and treatment of patients with congestive heart failure could have a favorable impact on the costs associated with the disease. B-type natriuretic peptide (BNP) is synthesized in the cardiac ventricles, and its level correlates with left ventricular pressure, amount of dyspnea, and the state of neurohormonal modulation, thus making peptide the first potential "white count" for heart failure. Data indicate that serial point-of-care testing of BNP should be helpful in patients presenting to urgent care clinics with dyspnea. BNP may also serve as a screen for patients referred for echocardiography. A low BNP level makes left ventricular dysfunction (both systolic and diastolic) highly unlikely. BNP may also provide an effective means of improving in-hospital management of patients admitted with decompensated congestive heart failure. In some cases, BNP level observations may obviate the need for invasive hemodynamic monitoring and, when such monitoring is used, may help tailor treatment of the decompensated patient. Finally, the role of BNP in the outpatient cardiac or primary care clinic may be one of critical importance in titration of therapies as well as in assessing the state of neurohormonal compensation of the patient. PMID- 11420772 TI - Health-related quality of life measurement in heart failure: challenges for the new millennium. AB - Health-related quality of life (HRQL), representing a patient-driven end point, has been increasingly emphasized in randomized clinical trials of new heart failure therapies. Measurement of HRQL depends on the use of validated instruments, with attention paid to the timing of administration and analysis of data in the context of conventional morbidity and mortality end points. In a review of HRQL measurement in heart failure drug trials published from 1966 to 1999, we found that important data, such as the number of participating subjects, are often lacking. HRQL is analyzed as a stand-alone end point without consideration of the underlying clinical trajectory of the disease. Improvements in trials methodology are warranted if quality-of-life data are to be meaningful in the determination of drug efficacy in heart failure. PMID- 11420773 TI - The prevention of osteoporosis using sequential low-dose hormone replacement therapy with estradiol-17 beta and dydrogesterone. AB - Low-dose hormone replacement therapy (HRT) in postmenopausal women may produce fewer side-effects but its efficacy in the prevention of bone loss and osteoporosis is not established. To address this we compared the effect of 1 mg estradiol-17 beta with a 2 mg dose, in combination with cyclical dydrogesterone, in the prevention of postmenopausal bone loss. We conducted a multicenter double masked prospective randomized, placebo-controlled study in 595 apparently healthy postmenopausal women randomized to either placebo, or continuous oral estradiol 17 beta 1 mg or 2 mg with sequential dydrogesterone for 2 years. The primary endpoint was the percentage change from baseline in bone mineral density (BMD) in the lumbar spine (LS) and femoral neck (FN) of actively treated groups compared with placebo. Women taking either 1 mg or 2 mg estradiol-17 beta showed a significant increase in BMD of the LS (mean +/- SD, 5.2 +/- 3.8% and 6.7 +/- 4.0% respectively, both p < 0.001) whilst BMD in the placebo group decreased (-1.9 +/- 4.0%). Increases were also observed in FN BMD in both treated groups (2.7 +/- 4.2% and 2.5 +/- 5.2% respectively, both p < 0.001) in contrast to the placebo group (-1.8 +/- 4.8%). The oldest women showed the greatest treatment response. One milligram estradiol-17 beta in combination with dydrogesterone is effective in conserving LS and proximal femur bone mass, both of which are clinically important sites of osteoporotic fracture, and is as effective as 2 mg in preventing FN bone. The lower dose of estradiol-17 beta is a particularly suitable treatment for osteoporosis management in older women since it should minimize side-effects and improve the acceptability of HRT. PMID- 11420774 TI - Bone mineral density at the hip predicts mortality in elderly men. AB - Low bone density as assessed by calcaneal ultrasound has been associated with mortality in elderly men and women. We examined the relationship between bone density measured at the hip and all cause and cardiovascular mortality in elderly men. Men aged 65-76 years from the general community were recruited from general practices in Cambridge between 1991 and 1995. At baseline survey, data collection included health questionnaires, measures of anthropometry and cardiovascular risk factors, as well as bone mineral density (BMD) measured using dual energy X-ray absorptiometry. All men have been followed up for vital status up to December 1999. BMD was significantly inversely related to mortality from all causes and cardiovascular disease, with decreasing rates with increasing bone density quartile, and an approximate halving of risk between the bottom and top quartile (p < 0.002, test for trend all causes and p < 0.025, test for trend for cardiovascular deaths). In multivariate analyses using the Cox proportional hazards model, an increase of 1 standard deviation (0.144 g/cm2) in total hip bone density was significantly associated with an age-adjusted 0.77 relative risk (95% CI 0.66-0.91) for all-cause mortality and 0.76 relative risk (95% CI 0.62 0.93) for cardiovascular disease mortality. The association remained significant after adjusting for age, body mass index, cigarette smoking status, serum cholesterol, systolic blood pressure, past history of heart attack, stroke or cancer and other lifestyle factors which included use of alcohol, physical activity and general health status. Low bone density at the hip is thus a strong and independent predictor of all-cause and cardiovascular mortality in older men. PMID- 11420775 TI - Oral glucocorticoid-induced fall in cortical bone volume and density in postmenopausal asthmatic patients. AB - Despite a deepening understanding of the influence of glucocorticoids (GC) on trabecular bone, little is known about GC-induced cortical bone loss. To elucidate the mechanism of GC-induced loss of cortical bone strength with particular reference to cortical bone loss, changes in cortical density, relative cortical volume, and the Strength Strain Index (SSI) based on biomechanical analyses of the geographic distribution of cortical bone material were measured. These parameters were compared, using peripheral quantitative computed tomography (pQCT), among the following age-matched groups: 68 postmenopausal asthmatic patients receiving high-dose oral GC in addition to inhaled GC (oral GC group), 68 postmenopausal asthmatic patients receiving only inhaled GC (inhaled GC group) and 69 postmenopausal controls without asthma or GC therapy (control group). Cortical bone mineral density (BMD) was measured, relative cortical volume was obtained by dividing the cortical area by the total bone area using pQCT (Stratec XCT960), and the Strength Strain Index (SSI) was calculated in the radius based on the density distribution around the axis. Spinal fracture was assessed on lateral radiographs. The number of vertebral fractures per patient correlated highly with cortical BMD, relative cortical volume and SSI values at the radius. The number of vertebral fractures per patient and the number of patients with fracture were similar between the control and inhaled GC group, both being significantly lower than those in the oral GC group. Total BMD, trabecular BMD, cortical BMD, relative cortical volume and SSI were similar between the first two, being significantly higher than in the last group. The slopes of cortical volume-density relationship, however, were identical among the three groups, indicating the persistence of cortical bone remodeling and a similar degree of calcification regardless of GC administration. PMID- 11420776 TI - Economic implications of hip fracture: health service use, institutional care and cost in Canada. AB - As the burden of illness associated with hip fracture extends beyond the initial hospitalization, a longitudinal 1 year cohort study was used to analyze levels of health service use, institutional care and their associated costs, and to examine patient and residency factors contributing to overall 1 year cost. Patients in the study were aged 50 year and over, and had been admitted to an acute care facility for hip fracture in the Hamilton-Wentworth region of Canada from 1 April 1995 to 31 March 1996. Health care resources assessed included initial hospitalization, rehospitalization, rehabilitation, chronic care, home care, long term care (LTC) and informal care. Regression analysis was used to determine the effects of age, gender, residence, survival and days of follow-up on 1 year cost. The mean 1 year cost of hip fracture for the 504 study patients was 26,527 Canadian dollars (95% Cl: $24,564-$28,490). One year costs were significantly different for patients who returned to the community ($21,385), versus those who were transferred to ($44,156), or readmitted to LTC facilities ($33,729) (p < 0.001). Initial hospitalization represented 58% of 1 year cost for community dwelling patients, compared with 27% for LTC residents. Only 59.4% of community dwelling patients resided in the community 1 year following hip fracture, and 5.6% of patients who survived their first fracture experienced a subsequent hip fracture. Linear regression indicated place of residence, age and survival were all important contributors to 1 year cost (p < 0.001). While the average 1 year cost of care was $26,527, the overall cost varied depending on a patient's place of residence, age, and survival to 1 year. Annual economic implications of hip fracture in Canada are $650 million and are expected to rise to $2.4 billion by 2041. PMID- 11420777 TI - Comparison of bone and total alkaline phosphatase and bone mineral density in postmenopausal osteoporotic women treated with alendronate. AB - Alendronate therapy in osteoporotic women decreases bone turnover and increases bone mineral density (BMD). Optimal patient management should include verification that each patient is responding to therapy. Markers of bone turnover and BMD have both been proposed for this purpose. We have investigated changes resulting from alendronate therapy with an enzyme immunoassay for bone alkaline phosphatase (BAP) and compared it with total alkaline phosphatase (TAP) and BMD of the lumbar spine, hip, and total body. Subjects were drawn from a multicenter randomized, placebo-controlled trial of alendronate in postmenopausal women with osteoporosis. BAP and TAP levels were measured at baseline and following 3, 6 and 12 months of therapy with either placebo (n = 180) or alendronate 10 mg/day (n = 134). All subjects also received 500 mg/day supplemental calcium. BMD was measured at baseline and following 3, 6, 12, 18, 24 and 36 months of therapy. To compare BAP, TAP and BMD at each site for identifying women that experienced a skeletal effect of alendronate, we calculated least significant change (LSC) values from the long-term intraindividual variability in each placebo-treated woman. Median levels of BAP decreased by 34%, 44% and 43% at 3, 6 and 12 months, respectively, in alendronate-treated women (p < 0.0001 compared with baseline and with placebo). These changes were significantly greater (p < 0.0001) than changes observed for TAP. Following 6 months of alendronate therapy, 90% of the women had experienced a decrease in BAP exceeding the LSC compared with only 71% for TAP. The greatest number of women similarly identified with BMD at any site (i.e. a gain in BMD exceeding the LSC) was 81% for spinal BMD at 36 months. All other sites were less than 70% at 36 months. Short-term changes in BAP and TAP were modestly associated with subsequent changes in BMD at all sites (Spearman's rho 0.22 to -0.52, p < 0.05). Compared with TAP and BMD, BAP testing rapidly and sensitively identified skeletal effects of alendronate thus enabling appropriate drug monitoring of osteoporotic women. Though BAP and TAP changes were modestly predictive of BMD changes, the value of the bone marker tests is their ability to detect rapidly a skeletal effect of therapy. PMID- 11420778 TI - Population bone mineral density measurements for Chinese women and men in Hong Kong. AB - The aim of the study was to establish population ranges of bone mineral density (BMD) for Hong Kong Chinese men and women for the Hologic QDR 2000 bone densitometer, to compare these values with the manufacturer's reference ranges, to compare these values with population ranges for women obtained for the Norland X26 bone densitometer, and to examine variations between the two densitometers. The subjects were 164 men aged 40-79 years and 436 women aged 20-89 years, who were all ethnic Chinese, recruited from volunteers, social centers for the elderly and general practice clinics. BMD in women began to decline rapidly between ages 50 and 79 years, averaging about 10% loss per decade from the young adult (20-29 years) mean. The percentage losses from young adult mean values in the spine, femroal neck, trochanter and total femur were 23%, 30%, 31% and 33%, respectively, from 20 to 79 years. In the ninth decade no further decrease in BMD occurred with the exception of a further 4% at the hip sites. In men, no decrease in spine BMD occurred between 40 and 70 years. Compared with BMD in the fourth decade, 10%, 13%, and 11% of BMD was lost at the femoral neck, trochanter and total femur, respectively, by the seventh decade. These values show differences compared with the manufacturer's reference ranges for Caucasians and Japanese. BMD values for the spine were comparable between Hologic and Norland densitometers, but Hologic values for femoral neck and trochanteric regions were lower than the Norland values. Data provided by this study may thus be used as normative values for the Hologic QDR2000 bone densitometer, instead of values provided by the manufacturer. BMD values at the hip sites are not interchangeable between Norland and Hologic bone densitometers, and estimation of numbers of the population with osteoporosis will depend on the model of densitometer used. PMID- 11420779 TI - Unbalanced diet to lower serum cholesterol level is a risk factor for postmenopausal osteoporosis and distal forearm fracture. AB - The purpose of this study was to assess whether dietary changes aimed at reducing serum cholesterol can increase the risk of osteoporosis (OP) and fracture. The study group consisted of 311 postmenopausal women with high serum cholesterol levels and following a diet low in dairy products (calcium intake estimated at less than 300 mg/day) for 27.3 +/- 29.1 months. This sample was compared with a case-control group of 622 healthy postmenopausal women paired for age and age at menopause and with a calcium intake estimated at more than 1 g/day. Bone mineral density was measured at the lumbar spine by dual-energy X-ray absorptiometry. Prevalence of OP was significantly higher in women with a low dairy calcium intake (42.1% vs 22.3%; p < 0.0001), as was the number of Colles' fractures occurring after menopause (4.5% vs 1.6%; p = 0.008). Multiple logistic regression analyses demonstrated that a diet low in dairy calcium was a risk factor for OP (OR = 2.52, 95% CI 1.84-3.45) and Colles' fracture (OR = 2.72, 95% CI 1.18-6.26). In the low dairy calcium group, diet duration significantly influenced the risk of OP (OR = 1.13, 95% CI 1.01-1.25 for 1 year of diet). No differences in further risk factors for coronary heart disease were found between the groups, but the proportion of women physically active was lower in the women with high serum cholesterol levels. A diet that severely limits calcium intake from dairy products in an attempt to correct raised serum cholesterol levels is a risk factor for postmenopausal OP and Colles' fracture. Dietary intervention methods to lower serum cholesterol in postmenopausal women should maintain an adequate calcium intake by providing calcium from low-fat dairy products or calcium supplements. PMID- 11420780 TI - Semiquantitative evaluation of prevalent vertebral deformities in men and their relationship with osteoporosis: the MINOS study. AB - Epidemiologic studies have shown a high prevalence of vertebral deformities in men without a steep increase with aging, suggesting that a substantial number of these deformities are not related to osteoporosis. To determine which vertebral deformities are likely to be osteoporotic fractures, we compared vertebral deformities and bone mineral density (BMD) in a cohort of 786 men aged 51-85 years (the MINOS study). Normal vertebral height ratios were defined in a group of 120 healthy men aged 21-50 years. We classified vertebral deformities by using the semiquantitative method described by Genant et al., which was slightly modified at the level of thoracic kyphosis (T6-T9). At that level, grade 1 wedge deformities were defined as a 25-30% decrease in anterior vertebral height and grade 2 by a 30-40% decrease. The same cutoff of 40% was used for grade 3 for all vertebrae from T4 to L4. BMD was measured with a Hologic 1500 device at the lumbar spine, hip and whole body and with an Osteometer DTX 100 device at the forearm. Z-scores were calculated in 10-year age groups. The prevalence of vertebral deformities increased significantly with age. After adjustment for age and body weight, BMD did not differ between those with and without vertebral deformities. In patients having grade 2 and 3 deformities, BMD was lower than in men having no deformities or only grade 1 deformities when adjusted for age and body weight. Using the age- and body-weight-adjusted test of linear trend for sextiles of BMD, prevalence of grade 2 and 3 vertebral deformities increased with a decrease in BMD at all the sites of measurement. Grade 1 deformities were not correlated with BMD at any site. Among 126 patients who had only grade 1 vertebral deformities, 32 deformities in 30 men were confirmed as vertebral fractures according to their morphology but their BMD did not differ from the nonfractured men. These findings were confirmed when vertebral deformities were measured by the conventional morphometric method in a subgroup of 131 men. Our data suggest that a cutoff of 30% for wedge deformities from T6 to T9 and of 25% for other deformities has a high specificity and a moderate sensitivity for identifying vertebral deformities related to low BMD in men. Grade 1 deformities are often either false positive or deformities related to nonosteoporotic disease of the spine. PMID- 11420781 TI - Hip fracture incidence rates in Singapore 1991-1998. AB - In this population-based study, we determined the incidence rates of hip fracture among Singapore residents aged 50 years and above. Information was obtained from a centralized database system which captured admissions with the primary diagnosis of a closed hip fracture (ICD-9 codes 820, 820.0, 820.2 and 820.8, n = 12,927) from all health care establishments in the country from 1991 to 1998 inclusive. After removing duplicates, hospital transfers, readmissions and non acute care admissions, the total number of hip fractures was 9406. Based on the national population census 1990 (n = 464,100) and yearly population estimates, the age-adjusted hip fracture rates for 1991-1998 (per 100,000) were 152 in men and 402 in women. This was 1.5 and over 5 times higher than corresponding rates in the 1960s. From 1991 to 1998, these hip fracture rates tended to increase by 0.7% annually in men and by 1.2% annually in women. Among the three major racial groups, in men, the Chinese had significantly higher age-adjusted hip fracture rates (per 100,000): 168 (95% confidence interval (CI) 158-178) compared with 128 (95% CI 105-152) for Indians and 71 (95% CI 54-88) for Malays. A similar pattern occurred in women: 410 (95% CI 395-425), for Chinese compared with 361 (95% CI 290-432) for Indians and 264 (95% CI 225-303) for Malays. Since the 1960s, the main increases in hip fracture rates have been seen in the Chinese and Malays, with the rates in Indians appearing to decrease. Hip fracture incidence rates in Singapore have risen rapidly over the past 30-40 years, particularly in women, and are now among the highest in Asia. Significant racial differences in hip fracture rates occur within the same community. Time trends in hip fracture rates differed between races. PMID- 11420782 TI - The effect of season and vitamin D supplementation on bone mineral density in healthy women: a double-masked crossover study. AB - Vitamin D status is known to be an important determinant of bone mineral density (BMD). There is a significant seasonal variation in serum vitamin D, and some studies have reported an associated seasonal variation in BMD. The present study was devised to investigate whether a seasonal variation in BMD could be detected in healthy normal subjects, along with associated variations in serum parathyroid hormone (PTH), intestinal calcium absorption and biochemical markers of bone turnover. A second aim was to investigate whether, if such variations were identified, they could be suppressed by vitamin D supplementation. The subjects were 70 healthy female volunteers (mean age 47.2 years, range 24-70 years) recruited into a double-masked crossover study and followed over 2 years. During the first year 35 subjects received a daily oral supplement containing 800 IU (20 micrograms) cholecalciferol (group 1) and 35 subjects received a placebo preparation (group 2). During the second year the treatment each group received was reversed. Lumbar spine (L1-L4), left proximal femur and total body BMD were measured by DXA at 3-month intervals. Serum 25-hydroxyvitamin D (25-OHD), serum PTH, bone markers (bone-specific ALP (BSAP) and urinary crosslinks (DYPD/creatinine)) and calcium absorption were also measured at each visit. Cholecalciferol treatment increased serum 25-OHD by 25.4 nmol/l (p < 0.001), while a reciprocal decrease in serum PTH of 6.6 ng/l (p = 0.011) was seen in subjects in the lowest quartile of baseline serum 25-OHD. The treatment had no significant effect on spine, femur or total body BMD, calcium absorption or bone markers. When Fourier analysis was used to analyze the data for seasonal effect (defined as twice the amplitude of the 1-year period variation) a highly significant effect for 25-OHD of 18 nmol/l (p < 0.001) was found. However, no effect was found for BMD, PTH, calcium absorption or bone markers. The analysis set a 95% confidence limit to the seasonal effect of less than 0.6% for spine, total hip and total body BMD. It was concluded that in the population of healthy women studied there was no evidence of seasonal variation in spine, femur or total body BMD, serum PTH, calcium absorption or bone markers. Vitamin D supplementation was found to have no effect on BMD. PMID- 11420783 TI - The Sp1 COLIA1 gene polymorphism, and not vitamin D receptor or estrogen receptor gene polymorphisms, determines bone mineral density in postmenopausal Greek women. AB - Several genetic polymorphisms are implicated as determinants of bone mineral density (BMD) in postmenopausal women. These include the Sp1 polymorphism of the collagen type I alpha 1 (COLIA1) gene, the FokI and BsmI polymorphisms of the vitamin D receptor (VDR) gene, and the PvuII and XbaI polymorphisms of the estrogen receptor (ER) gene. The relative importance and the independence of these genetic effects have not been studied simultaneously in the same population. We evaluated the effects of these polymorphisms on lumbar spine BMD among 154 postmenopausal Greek women. BMD tended to differ across Sp1 genotypes (mean 0.842 g/cm2 in SS, 0.851 g/cm2 in Ss, 0.763 in ss, age-adjusted p = 0.056), mostly because ss homozygotes had lower BMD (p = 0.018 compared with SS and Ss). No other polymorphisms were associated with BMD in this population (p = 0.53 for FokI, p = 0.94 for BsmI, p = 0.80 for PvuII, p = 0.91 for XbaI). In multivariate modeling, the effect of ss homozygosity was clinically and statistically significant (-0.105 g/cm2, p = 0.013) after adjusting for age, weight, height, hormone replacement use, and the other four polymorphisms. None of the other four polymorphisms was retained as an independent predictor of BMD in a backward elimination model and no significant synergistic effects were observed when gene interactions were tested. When all five polymorphisms are considered simultaneously, the Sp1 COLIA1 polymorphism seems to have the most unequivocal effect on BMD, at least in postmenopausal women. PMID- 11420784 TI - Osteoporosis and apolipoprotein E genotype in older adults: the Rancho Bernardo study. AB - Recent studies reported an association between apolipoprotein E (ApoE) 4 and osteoporosis. We examined the association of ApoE 4 genotype with bone mineral density (BMD), bone loss and fracture risk in 596 men and 332 community-dwelling women aged 45-95 years. Women were postmenopausal and not using estrogen. At the baseline visit, BMD was measured at the ultradistal and midshaft radius using single photon densitometry, and at the hip and lumbar spine using dual-energy X ray absorptiometry. Hip and lumbar spine BMD levels were remeasured 4 years later. Self-reported fractures were confirmed by radiology reports in 95% of cases. ApoE allele distribution did not vary by age; 25% of men and 20% of women had one ApoE 4 allele. There were no differences in BMD at the lumbar spine, total hip, ultradistal or midshaft radius in men or women with the ApoE 4 allele compared with men or women without the ApoE 4 allele. After an average 4 year interval, there were also no differences in the annualized percent change in BMD at the hip or lumbar spine in men or women with or without an ApoE 4 allele. One or more clinical fractures were reported by 55 men and 109 women. Fewer, not more, clinical fractures were reported in men and women with an ApoE 4 allele; these differences were not statistically significant (p = 0.21 and p = 0.62, respectively). These data do not support the hypotheses that there is an association between ApoE genotype and BMD, bone loss or osteoporotic fractures in older community-dwelling men or women. PMID- 11420785 TI - Can proprioception really be improved by exercises? AB - There is little question that ankle disc training can improve ankle muscle motor performance in a unipedal balance task, most likely through improved strength and coordination [62] and possibly endurance. How much of the observed improvement in motor performance is due to improved ankle proprioception remains unknown. We have reviewed a number of theoretical ways in which training might improve proprioception for moderately challenging weight-bearing situations such as balancing on one leg. Although the relevant experiments have yet to be performed to test this hypothesis, any improvement would theoretically help to reduce injuries at these moderate levels of challenge. We question, however, whether these exercises can ever improve the reactive response required to prevent injury under the most challenging time-critical situations. If confirmed, this limitation needs to be acknowledged by authors and practitioners alike. Alternative protective strategies for the most challenging time-critical situations should be sought. We conclude that, despite their widespread acceptance, current exercises aimed at "improving proprioception" have not been demonstrated to achieve that goal. We have outlined theoretical scenarios by which proprioception might be improved, but these are speculative. The relevant experiments remain to be conducted. We argue that even if they were proven to improve proprioception, under the best circumstances such exercises could only prevent injury under slow to intermediate rate provocations to the joint musculoligamentous complex in question. PMID- 11420786 TI - Reconstruction of the anterior cruciate ligament with the iliotibial band autograft in patients with chronic knee instability. AB - We performed combined internal and external anterior cruciate ligament (ACL) reconstruction with the iliotibial band autograft in 169 consecutive patients with chronic ACL insufficiency who were followed up for 24-61 months. Of these, 155 (91%) agreed to an additional independent observer follow-up after 24-92 months. Eight patients (5%) had sustained a rerupture/elongation of the graft and were operated on again; nine (6%) had sustained a tear of the contralateral ACL. Knee function and activity increased after the reconstruction. Lysholm scores improved from median 81 preoperatively to 99 at follow-up and Tegner scores from median 4 to 7. At follow-up 97 (71%) were active at the same level as prior to injury. In 17 of the 40 patients (12%) dropping to a lower activity level this was due to knee problems. The side-to-side difference in anterior-posterior knee laxity was more than 3 mm in 18 knees (13%) and more than 5 mm in 3 knees (2%). Including eight reruptures, this results in a "stability" failure rate of 8.8%. The overall IKCD rating showed normal knee function in 88 (73%) and nearly normal knee function in 30 (25%). Anterior knee pain was present in 14 (10%) of the patients at follow-up. Patients with isolated ACL injury had higher Lysholm scores and Tegner scores than patients with associated injuries. No clinical signs of varus knee development were seen. Of the 155 patients 94% would have the procedure repeated if necessary with the knowledge that they have today. The combined internal and external iliotibial band procedure can restore knee stability and function in the majority of chronic ACL-insufficient knees. PMID- 11420787 TI - Activity-related knee injuries and pain in athletic adolescents. AB - By collecting data from 45 students at a ski high school, we found that a total of 73% of the students reported activity-related pain/injuries of the knee. Sixty one percent had overuse injuries, 27% malalignment, and 12% had indistinct knee pain. Females suffered more knee pain/injuries (88%) than males (57%). Significantly higher Q-angle degrees were recorded for females (16) than for males (10). "Jumper's knee" was found in all competitive students with a KT manual maximum difference (MMD) of 3 mm or more (mean 4 mm), with a hard endpoint, whereas this was less common among the other competitive students (P < 0.05). The students were given counselling about training and physiotherapy. In the follow-up study 1 year later, a significant reduction of knee pain/overuse injuries, from 73% to 35%, was recorded. This may be related to better equipment, the development of techniques, and training of the muscles. A high volume of training and knee instability, with MMD of 3 mm or more, seemed to be correlated with an increased risk for "jumper's knee" and, possibly, for skiing injuries. By identifying those at increased risk, preseason recommendations can be made and ski injuries may be prevented. PMID- 11420788 TI - Patellar fractures associated with medial-third bone-patellar tendon-bone autograft ACL reconstruction. AB - Patella fractures following anterior cruciate ligament (ACL) reconstruction are a recognized but rarely reported complication. To our knowledge, 24 reports of patella fractures after ACL reconstruction using the central-third patella-tendon autograft have been reported in the literature. Patellar fractures associated with the use of the medial-third bone-patellar tendon-bone autograft have not been reported. This article describes four cases of patellar fractures in 478 ACL reconstructions between 1992 and 1999, using the medial third of the patellar tendon graft. All of them were transverse fractures of the patella but only one was displaced. All patients suffered local injury to the donor knee between 2 and 4 months postoperatively. No significant differences in the final outcome were noticed between the cases complicated with patellar fracture and those with uncomplicated ACL reconstructions. PMID- 11420789 TI - Patello-femoral tracking in the weight-bearing knee: a study of asymptomatic volunteers utilising dynamic magnetic resonance imaging: a preliminary report. AB - Normal patello-femoral tracking is not well defined, and conventional radiological techniques do not allow imaging in the physiological, weight-bearing stance. A vertical-access open configuration magnetic resonance scanner allows imaging of patello-femoral tracking during weight-bearing and through a wide range of knee flexion. We imaged 40 asymptomatic knees in this way, producing axial scans which were analysed qualitatively and quantitatively using sulcus angle, congruence angle, lateral patello-femoral angle and patellar centralisation, to assess patellar tilt and displacement. Mild lateral tilting in hyperextension with the quadriceps relaxed was seen, but quantitative assessment of this was impeded by internal rotation of the femur in extension. One-half of the knees were slightly laterally displaced in hyper-extension, becoming central during the first 30 degrees of knee flexion. During passive flexion of the knee in a seated position, fewer knees were laterally tilted or displaced, and no consistent change was seen during flexion. These results indicate that mild lateral tilting and displacement can be normal phenomena in the weight-bearing knee in early flexion and should not necessarily be taken as evidence of abnormal tracking in symptomatic patients. Lateral to medial movement of the patella occurs during normal knee flexion. In addition, imaging in the weight-bearing knee can provide valuable information not gained by imaging during passive knee flexion. PMID- 11420790 TI - Tendon lengthening repair and early mobilization in treatment of neglected bilateral simultaneous traumatic rupture of the quadriceps tendon. AB - Bilateral simultaneous traumatic rupture of the quadriceps tendon is a rare injury that is most frequently seen in elderly patients with predisposing diseases such as gout, hyperparathyroidism and diabetes. Delay in diagnosis is not uncommon. One of the main problems in treatment is loss of motion, especially flexion, after surgical repair. We report a case that was diagnosed 5 months after the trauma and was treated by Scuderi's tendon lengthening technique. Range of-motion exercises were started early without using the generally recommended 4 6 weeks of immobilization in plaster cylinder or knee brace. Five years of follow up showed full range of motion in both knees with sound tendons. Stable fixation makes starting early motion and accelerated rehabilitation feasible and thus the most common complication, loss of motion, is prevented. PMID- 11420791 TI - Arthroscopic reduction and internal fixation (ARIF) of lateral tibial plateau fractures. AB - Arthroscopic reduction and internal fixation of tibial plateau fractures has been well documented over the last 15 years. Better visualization, less traumatic surgery, reconstruction of accompanying injuries and early mobilization have encouraged arthroscopic surgeons to use this technique more widely. With experience, surgeons became more comfortable in using arthroscopy-assisted surgery not only in split fractures of the lateral tibial plateau (41-B1) but also in more complex tibial and femoral fractures. In this study, 31 patients with intra-articular fractures of the lateral tibial plateau were operated on between 1991 and 1996. The fractures were classified as follows: x12 AO-type B1, x7 B2, x10 B3, x2 C3. Diagnostic arthroscopy and repair of soft tissue injuries was followed by arthroscopic reduction in all patients and supplemented with internal fixation in 29 patients. Postoperatively, an aggressive physical therapy regimen was performed with immediate mobilization and early partial weight bearing. All patients were examined for follow-up after 15-32 months (mean 25.1). In all cases the fractures were stable and allowed full weight-bearing. At follow up, 25 patients had anatomic reduction. Anatomical reduction cannot be restored in all cases of open reconstruction because of cartilage defects. Fracture reduction using arthroscopic techniques is a suitable alternative for joint fracture repair because additional soft tissue damage can be minimized. PMID- 11420792 TI - Extensive osteolytic cystlike area associated with polyethylene wear debris adjacent to an aseptic, stable, uncemented unicompartmental knee prosthesis: case report. AB - We present the case of a patient who after uncemented unicompartmental knee arthroplasty developed a large osteolytic cystlike area in the lateral aspect of the tibial metaphysis, contralateral to a well-fixed tibial component at revision surgery. The lesion contained fibrotic soft tissue, evidence of a foreign-body giant cell reaction and polyethylene particles, but no metal wear debris, infection, or malignancy. This case demonstrates that there is a direct communication between the joint cavity and the cyst. PMID- 11420793 TI - 360 degrees arthroscopic capsular release in patients with adhesive capsulitis of the glenohumeral joint--indication, surgical technique, results. AB - Adhesive capsulitis of the glenohumeral joint is said to be a self-limiting process. However, in some patients the disease can last much longer than 1 year, which may lead patients to more invasive treatment than merely undergoing physiotherapy. Other patients do not accept this severe limitation and choose treatment options that restore the range of motion (ROM) more rapidly. Conventional open release techniques generally improve motion but involve extensive dissection. The purpose of this study was to develop a safe and reproducible technique of arthroscopic capsular release (ACR) and to present the results of this technique in the clinical situation. The technique for ACR was first defined in a cadaver study and then applied in 28 patients with primary adhesive capsulitis of the glenohumeral joint. The patients were selected for the arthroscopic release when conservative therapy had failed for at least 6 months. All of the patients had a global loss of shoulder motion and had motion restored with a combined anterior, posterior, superior, and inferior release of the of the capsule (360 degrees release). Additionally, in all patients synovectomy with electrocautery was performed. We documented the ROM in the different planes as well as the Constant score. The Constant score improved a mean of 41 points. Range of motion for all planes significantly improved (P < 0.01). Abduction improved from 75 degrees preoperatively to 165 degrees intraoperatively; 6 weeks after surgery, mean abduction was 168 degrees and at the time of follow-up it was 167 degrees. Mean external rotation in adduction improved from 3 degrees preoperatively to 75 degrees intraoperatively. After 6 weeks, the mean external rotation in adduction was 72 degrees and at the time of follow-up the external rotation reached 76 degrees. Mean external rotation in abduction improved from 4 degrees preoperatively to 81 degrees intraoperatively, 80 degrees after 6 weeks and 85 degrees at the time of the last follow-up. Internal rotation in abduction was 17 degrees preoperatively. Intraoperatively, mean internal rotation was 59 degrees. An angle of 58 degrees was documented at 6 weeks follow-up, and at the last follow-up an angle of 63 degrees was documented. No postoperative lesion of the axillary nerve was present. We concluded that arthroscopic capsular release is a reliable method for restoring motion with minimum morbidity in carefully selected patients. When performing an ACR the incision of the glenohumeral joint capsule should be undertaken at the glenoidal insertion in the abducted and external rotated shoulder. PMID- 11420794 TI - The intra- and inter-observer reliability of the Samilson and Prieto grading system of glenohumeral arthropathy. AB - Severity of glenohumeral arthropathy following shoulder instability is often graded radiologically on the basis of osteophyte size according to Samilson and Prieto. Intra- and inter-observer reliability of this popular grading system was determined using standard radiographs of 20 arthritic cadaveric humeri. Using two observers on two occasions, reliability of the grading system using the antero posterior radiograph varied between fair and poor (kappa coefficients 0.07-0.33). This study shows that the Samilson and Prieto grading of glenohumeral arthropathy is unreliable and, in clinical use, when magnification and rotation is less easy to control, it is likely to be even more so. Another validated classification system must therefore be found. PMID- 11420795 TI - EJPH Policy Forum: risk adjustment strategies in three social health insurance countries. PMID- 11420796 TI - Body height, birth cohort and social background in Finland and Sweden. AB - BACKGROUND: Poor childhood living conditions are associated with short stature. Before the Second World War Finland had much lower living standards than Sweden, but this gap had largely disappeared by the 1970s. Body height differences were examined by birth cohort, economic difficulties in childhood and adult socioeconomic position in Finland and Sweden. METHODS: Two nationally representative data sets were used (n = 7,300 in Finland and n = 4,551 in Sweden). Three indicators of social background were included, i.e. economic difficulties in childhood, education and occupational class. The methods used were direct age-standardisation, index of dissimilarity and regression analysis. RESULTS: In the cohort born in 1920-1929 body height was taller in Sweden (175.8 cm among men and 163.7 cm among women) than in Finland (173.9 and 161.2 cm respectively). Body height by birth cohort increased faster in Finland, with the result that, in the cohort born in 1960-1969, the gap between the countries had narrowed to 0.8 cm among men and 0.3 cm among women. Body height differences by social background were larger in Finland than in Sweden. Socioeconomic body height differences have remained largely stable over the birth cohorts in both countries. CONCLUSIONS: The results suggest that differential economic development is partly seen in the narrowing of body height differences between Finland and Sweden. However, socioeconomic differences in body height have remained largely similar over the birth cohorts studied and between Finland and Sweden. PMID- 11420797 TI - Social influences and low leisure-time physical activity in young Danish adults. AB - BACKGROUND: Many youths and adults maintain a sedentary life style and a better understanding of the factors which influence physical activity is needed. This study analyses the association between social factors and low leisure-time physical activity in young adults. METHODS: Three hundred and seventeen children aged 6-18 years participated in a health survey in 1979 and again 13 years later. In this cohort the influence of family factors during childhood on the subsequent risk of low leisure-time activity was analysed using multiple logistic regression. RESULTS: Young women were significantly less physically active during leisure time if over 25 years of age, poorly educated and smokers. In multivariate analysis, childhood smoking was the only variable significantly related to low leisure-time physical activity (OR = 5.6 and 95% CI: 1.4-23.6) in young women. In young men, low leisure-time physical activity was associated with parental work activity. Men whose parents had reported high physical activity during work at the examination in 1976-1978 were less often physically inactive during leisure time (OR = 0.2 and 95% CI: 0.1-0.6) at the follow-up in 1992. CONCLUSIONS: Young smoking girls seem to be a key group in the prevention of both smoking and a sedentary life style. PMID- 11420798 TI - Health-related mobility, health inequalities and gradient constraint. Discussion and results from a Norwegian study. AB - BACKGROUND: Recent studies have argued that health-related mobility does not widen social class health differentials, but rather moderates them. This is termed gradient constraint. This paper examines gradient constraint from a theoretical and empirical angle. METHODS: How health-related mobility influences social class health differentials is discussed using hypothetical models. In a Norwegian survey with data on intergenerational mobility (N = 1,853 males aged 30 69 years), mean health and height values for different subsections of the sample were analysed. RESULTS: When initial social class health differences are large and mobility widespread, health-related mobility may lead to smaller differentials, but the result depends on how closely mobility varies with health. This empirical study found non-significant increases in height and health differentials from class of origin to class of destination. The interpretation has to consider effects of both social mobility and social causation. Health is measured in the post-mobility situation and the effects of social mobility and social causation are difficult to separate from each other for most of the health indicators analysed. However, this limitation does not apply to height which is not influenced by social causation during adulthood. In these data mobility did not reduce the height differential. CONCLUSION: Health-related mobility can either lead to smaller or larger social class health differentials. The specific effects of social mobility cannot be determined without knowing how social causation has interfered. The intergenerational mobility process analysed in this paper does not show gradient constraint as regards the height differential between the worker and higher occupational categories. PMID- 11420799 TI - Body mass index and weight change attempts among adult women. The Norwegian Women and Cancer Study. AB - BACKGROUND: Based on concern about the widespread increase in being overweight and obesity along with a substantial demand for leanness, we wanted to estimate body mass index (BMI) and the prevalence of weight change attempts in a population-based sample of Norwegian women. Furthermore, we wanted to examine how weight loss attempts are related to BMI and to age, socioeconomic status, reproductive factors, lifestyle and diet. METHODS: A nationwide, cross-sectional study applying a mailed questionnaire was used. Out of a random sample of 20,000 women aged 45-69 years 10,249 women participated. RESULTS: Based on self-reported data, the mean BMI was 24.6 kg m-2 and 40% of the women had a BMI of > or = 25 kg m-2. More than 50% of the women were trying to lose weight and weight loss attempts were very strongly associated with BMI. Age, education, income, smoking status and perception of diet's importance to health were also significant predictors of weight loss attempts. The effect of age, education and income on weight loss attempts was modified by the level of BMI. Women trying to lose weight reported a different diet than those not trying to lose weight, irrespective of BMI. CONCLUSION: A large proportion of middle-aged women are trying to lose weight. BMI is predominant in explaining weight loss attempts. After adjusting for BMI, age, lifestyle and socioeconomic status also contribute to explaining weight loss attempts. PMID- 11420800 TI - Is there equity in access to health services for ethnic minorities in Sweden? AB - BACKGROUND: This paper addresses the extent to which equity of treatment according to need, as defined by self-reported health status, is received by members of ethnic minorities in Swedish health services. METHODS: The study was based on a multivariate analysis of cross-sectional data from the Swedish Survey of Living Conditions and Immigrant Survey of Living Conditions in 1996 on use of health services, morbidity and socioeconomic indicators. The study population consisted of 1,890 Swedish residents aged 27-60 years born in Chile, Poland, Turkey and Iran and 2,452 age-matched, Swedish-born residents. MAIN RESULTS: Residents born in Chile, Iran and Turkey were more likely to have consulted a physician during the 3 months prior to the interview compared to Swedish-born residents; odds ratios (ORs) 1.4 (95% CI: 1.2-1.7), 1.3 (95% CI: 1.1-1.7) and 1.5 (95% CI: 1.3-1.9) respectively. The higher consultation rate in these ethnic minorities was primarily explained by a less satisfactory, self-reported health status compared to Swedish-born residents. Thirty-eight percent of the minority study groups reported exposure to organised violence in their country of origin, which was associated with a higher level of use of consultations with a physician (OR 1.3, 95% CI: 1.1-1.6). CONCLUSIONS: This study did not indicate any gross pattern of inequity in access to care for ethnic minorities in Sweden. Systems for allocating resources to health authorities need to consider the possibility that ethnic minorities in Sweden and in particular victims of organised violence, use health services more than is suggested by socioeconomic indicators only. PMID- 11420801 TI - Effects of peer-led AIDS education aimed at Turkish and Moroccan male immigrants in The Netherlands. A randomised controlled evaluation study. AB - BACKGROUND: An evaluation study was conducted in The Netherlands into acquired immune deficiency syndrome (AIDS) education for immigrants given in their native language by peers. Turkish and Moroccan men were trained to educate people from their own ethnic group. The effect of peer education on the perceived threat of AIDS and beliefs about condom use were studied. METHODS: Places where male immigrants met, i.e. coffee houses, mosques and bars, were matched and randomly assigned to experimental and control groups. The experimental group filled out a short questionnaire at the end of the education session (post-test), whereas the control group was pre-tested and had the opportunity of following the AIDS education after participation in the questionnaire. RESULTS: Using multilevel logistic regression analysis, an effect could be established on misunderstandings regarding human immunodeficiency virus (HIV) transmission (OR = 5.9 and 95% CI: 2.3-15.3) and risk appraisal for HIV infection (OR = 2.9 and 95% CI: 1.3-6.3). The perceived benefits of the protective effect of condom use were affected in men 30 years and older, the perceived barrier of diminished satisfaction if using condoms was changed among unmarried men, condom self-efficacy was affected in men who valued peer education as important and an effect on intention to use condoms was found among Moroccans. CONCLUSION: Continuation of peer-led AIDS education for immigrants and adaption of the message to the needs of specific target groups is recommended. PMID- 11420802 TI - Social democratic government and spatial distribution of health care facilities. The case of hospital beds in Germany. AB - BACKGROUND: In this paper, the hypothesis that the spatial distribution of hospital beds is more even in countries with socialist or social democratic governments than in countries with conservative or Christian democratic governments was tested. To avoid the confounding influences of historical and institutional differences between countries, we used the Federal Republic of Germany as a case study. The German federal states have their own governments who play an important role in creating structures for the planning of hospital facilities. METHODS: The test of the hypothesis was largely quantitative. At the level of federal states the rank correlation was computed between the weighted number of years of left-wing government participation and the coefficient of variation in the number of hospital beds per 1000 inhabitants. In addition to this, the hospital plans of two federal states were studied. RESULTS: The hypothesis was supported by the data, showing a positive association between the number of years of left-wing government participation and regional variation in the number of hospital beds. A comparison of the hospital plans of two contrasting federal states showed less government interference in hospital planning in the state with a tradition of right-wing government. CONCLUSION: There seems to be a relation between left-wing government participation in West German states and a more equal distribution of the number of hospital beds per 1,000 inhabitants. PMID- 11420803 TI - Length of stay. How short should hospital care be? AB - BACKGROUND: Reducing length of hospital stay (LOS) is a policy aim for many health care systems and is thought to indicate efficiency. METHODS: A MedLine search was undertaken for articles relating to 'LOS', 'early discharge' or 'patient discharge' between 1983 and 1997 and a selective search was undertaken for material published before 1983. RESULTS: Routine data showed that there were variations in LOS between countries, regions and hospitals. The trends in LOS showed a decrease over time in all regions. Research consistently fails to show an adverse effect on health outcomes of reducing LOS, but there may nevertheless be an ethical or moral minimum LOS. Two recent examples illustrate this. There has been an outcry at some ultrashort stays, for example 'drive-through mastectomy' and 'lunchtime abortion' and these are discussed in the review. CONCLUSIONS: There are a number of reasons for the perceived lack of relationship between LOS and health outcomes. Clearly reducing days of care at the low intensity end of a hospital stay may not necessarily affect health outcomes. There is a case to be made for tailoring care more exactly to an individual's needs by looking at the actual components of care rather than the place of care- within or outside hospital walls. PMID- 11420804 TI - Public health training in Europe. Development of European masters degrees in public health. AB - BACKGROUND: Changing political and economic relations in Europe mean that there are new challenges for public health and public health training. There have been several attempts to develop training at the master's level in public health which is focused on meeting the new needs. These have failed due to being too inflexible to allow participation by schools of public health. METHODS: A project funded by the European Union involving public health trainers has developed a new approach which allows participating schools to retain their national differences and work within local rules and traditions, but which aims to introduce the European dimension into public health training. This paper reports the conclusions of this project. CONCLUSIONS: A network of schools wishing to develop European Master's degrees is being established and other schools offering good quality programmes will be able to join. PMID- 11420805 TI - Risk structure compensation in Germany's statutory health insurance. AB - In 1996 insurees in Germany's statutory health insurance system were given a right to choose their sickness fund. To ensure that all funds had an equal starting position, a risk structure compensation scheme based on income and average expenditure by age and sex was introduced. From an analysis of expenditure and transfers, data on sickness fund membership and a published survey, the following effects can be identified: sickness funds merged, with a reduction in number from 1,221 to 420 between 1993 and 2000; the risk compensation scheme narrowed differences in contribution rates; insurees left the more expensive funds for cheaper ones; and increasing transfer sums indicate further risk segregation. Thus, the compensation mechanism will have to be retained permanently, although modifications are likely. PMID- 11420806 TI - Implementing prospective budgeting for Dutch sickness funds. AB - Most if not all social policies entail redistribution of scarce public resources from central government to regional and local authorities, to individual citizens or non-government agencies. Governments use a wide variety of instruments to allocate public funds, including direct state provision of subsidies and goods and services, setting budgets at different levels, and regulation of social insurance schemes. Most industrialised countries have developed budget models based on implicit or explicit allocation criteria. Governments usually start by determining global budgets for an entire category of public spending and then specifying the amounts allocated for categories of spending, and next, the budgets for individual agencies. Within such a 'cascading' model, the lower level budgets may be more controversial than the global budgets, as they directly affect the amounts available to individual actors in the system, e.g. hospitals or health insurance agencies. Setting budgets not only shifts decision-making authority but also financial risks from the central government to decentralised actors. The introduction of the prospective budgeting model for the Dutch sickness funds illustrates why determining budgets is not merely a matter of choosing objective allocation criteria, but also, of interaction between state and stakeholders. In the typical Dutch neocorporatist policy arena, where organised interests share responsibilities with government for the shaping and implementation of social policies, the health insurance agencies actively participated in the development of the budget model. PMID- 11420807 TI - Risk-adjusted capitation. The Israeli experience. AB - As in a number of countries during the 1990s, Israel's health system has been undergoing structural reform based on public contracting and regulated markets. The main element of the reform was the enactment of the National Health Insurance Law (NHI), which went into effect on 1 January 1995. According to the Law, the sick funds receive risk-adjusted capitation payments, which place them fully at risk for the cost of supplying a legally mandated basket of health benefits. The paper analyses the effects of the NHI on the Israeli competitive health insurance market and discusses the major policy issues facing the Israeli system. PMID- 11420808 TI - A comparison of high-risk sexual behaviour and HIV testing amongst a bar-going sample of homosexual men in London and Edinburgh. AB - BACKGROUND: This study compared high-risk sexual and HIV testing behaviour amongst homosexual men recruited from gay bars in London and Edinburgh. METHODS: A cross-sectional survey monitoring high-risk sexual and HIV testing behaviour using a self-completed questionnaire was conducted in November and December 1996. RESULTS: Two thousand, three hundred and ninety-seven questionnaires were returned (1,366 recruited in London and 1,031 in Edinburgh), with a response rate of 77%. A larger proportion of men surveyed in London had had unprotected anal intercourse (UAI) with one or more male partners in the previous year (35%) than in Edinburgh (30%). Men recruited in Edinburgh were less likely to have had an HIV test (54%) than men in London (63%). In both surveys, 25% of men who reported UAI with partners of the same HIV status as themselves also reported never having had an HIV test. CONCLUSIONS: The observed dissimilarities in the HIV epidemic in the two cities may be accounted for by the differences in self-reported high-risk sexual and HIV testing behaviours between the two populations. A large proportion of men in both cities continue to engage in high-risk sexual behaviour suggesting continued transmission of HIV in these populations. Thus, there is a continued need for innovative and relevant health promotion amongst homosexual men in the UK. PMID- 11420809 TI - Does the liberalisation of abortion laws increase the number of abortions? The case study of Spain. AB - BACKGROUND: Over the course of the 1980s a public debate on abortion took place in Spain culminating in a more permissive social climate and, in 1985, the partial decriminalisation of abortion. Before this, women were forced to abort illegally or abroad in countries which had decriminalised abortions. The aim of this study is to present jointly the evolution of abortions in Spanish women in England and Wales between 1974 and 1995, The Netherlands between 1980 and 1995 and Spain since the start of the register in 1987 through to 1995 and to compare trends both before and after the law in Spain. METHODS: Incidence rates were calculated in each of the countries studied and the slopes of the curves for 1974 1984 and 1987-1995 were compared. Data were obtained from reports published by the offices of abortion surveillance in England and Wales, The Netherlands and Spain. RESULTS: The rates increased constantly throughout the study period. From 1974 to 1985, a total of 204,736 Spanish women aborted in England and Wales and The Netherlands. After the law was passed, 34,895 Spanish women had abortions in those countries over the period 1986-1995. During 1987-1995, 340,214 Spanish women terminated their pregnancies in Spain. The regression coefficients before and after the passing of the law were beta = 0.3538 (0.307-0.400) and beta = 0.319 (0.243-0.394) respectively; no difference was observed. CONCLUSIONS: During the study period a significant proportion of reproductive-aged Spanish women had abortions in England and Wales and The Netherlands. Decriminalisation has had no observed effect on the trends in abortion, but rather it has benefited Spanish women by making abortion available locally and, therefore, reducing the inequalities implied by lack of access to proper health care services. These data demonstrate the impact of the liberalisation of abortion on the trends of procedures performed in other countries. PMID- 11420810 TI - The prevalence of the use of androgenic anabolic steroids by adolescents in a county of Sweden. AB - BACKGROUND: The prevalence of the use of androgenic anabolic steroids has been poorly studied in Europe. This study was undertaken to examine the prevalence of the misuse--the non-medical use--of androgenic anabolic steroids among adolescents in a county of Sweden. METHODS: The total population of 16 and 17 year old male and female adolescents in a county on the south-west coast of Sweden was studied. The investigation was done by an anonymous multiple-choice questionnaire. The questionnaire was completed by 5,827 pupils and statistically analysed. The participation rate was 95%. RESULTS: Among male adolescents 16 and 17 years old, 3.6% and 2.8% had misused androgenic anabolic steroids, respectively. These male adolescents had also misused alcohol, growth hormones and narcotic drugs more than the steroid hormone non-users. Among female adolescents there was no recorded misuse of these drugs (0.0%). CONCLUSIONS: The misuse of androgenic anabolic steroids is a reality in both small and large municipalities in Sweden. The prevalence figures are higher among 16 year old compared to 17 year old male adolescents. There is an association between this drug misuse and other substance misuse such as narcotic drugs. Female adolescents do not misuse steroid hormones. The findings indicate the need for preventive work among male adolescents in order to induce adolescents not to start misusing androgenic anabolic steroids. PMID- 11420811 TI - A capture-recapture study of the prevalence and implications of opiate use in Dublin. AB - BACKGROUND: To date there have been no studies estimating the hidden prevalence of opiate use in Dublin. METHODS: A multisource enumeration followed by the application of the capture-recapture method with log-linear modelling including age and gender stratification to remove heterogeneity was implemented to provide an estimate of the unknown size of the opiate-using population. Two medical and one legal data sources were used. RESULTS: It was found that the ratio of known to unknown opiate users was 1:1.15 with a total of 13,460 (95% CI: 12,037-15,306) users estimated in Dublin in 1996. CONCLUSION: The findings of this study have important ramifications for service delivery. PMID- 11420812 TI - Litigation on behalf of victims of exposure to environmental tobacco smoke. The experience from the USA. AB - BACKGROUND: For a quarter of a century, lawsuits have been filed on behalf of individuals who have been harmed by exposure to environmental tobacco smoke (ETS). METHODS: A review of hundreds of legal cases where exposure to ETS was the basis for the lawsuit was undertaken. The history of ETS-based lawsuits was categorised and analysed. RESULTS: While ETS-based lawsuits have arisen in a variety of divergent settings, there have been an increasing number of instances in which individuals have achieved relief from exposure to ETS and/or monetary compensation for having been so exposed. Such successful outcomes make it easier for others to achieve similar results through litigation. CONCLUSION: Lawsuits will probably continue to be a viable remedy in assisting those being harmed by exposure to ETS, both-inside and outside the USA. PMID- 11420813 TI - Smoking and passive smoking in Estonia, Lithuania and Finland. Identifying target groups of tobacco policy. AB - BACKGROUND: The aim of this study was to identify population groups which are the most crucial as targets for anti-tobacco action. METHODS: A comparison was made of the prevalence and patterns of smoking and passive smoking in Estonia, Finland and Lithuania. RESULTS: Total exposure to smoking, both one's own and passive smoking, was more common in two Baltic countries than in Finland. In these Baltic countries passive smoking was notably common among women. In all countries the exposure was more prevalent among the younger and less educated, but no difference emerged between urban and rural areas. CONCLUSIONS: The following target groups were identified as priorities for anti-tobacco actions: i) men of all ages in Estonia and Lithuania to stop their already established tobacco use, ii) young women in all three countries to prevent their starting a career of tobacco use and iii) young and less educated women in Estonia and Lithuania to prevent passive smoking, i.e. their exposure to environmental tobacco smoke. PMID- 11420814 TI - Smoking and quitting with the aid of nicotine replacement therapies in the English adult population. Results from the Health Education Monitoring Survey 1995. AB - BACKGROUND: Many clinical trials have shown the usefulness of nicotine replacement therapies, family physician advice and other cessation aids in enhancing smoking cessation. However, many trials have taken place amongst selected patients and doctors. This paper assesses cessation amongst a representative general adult population in England in order to see whether such aids are as successful amongst more typical smokers. METHODS: Descriptive information on the pattern of smoking cessation aids use in the English population is presented for the first time. This is followed by multivariate probit analysis of the 'Health in England 1995' survey data set in order to predict successful smoking cessation as a function of several variables including use of nicotine gum and patches, willpower, advice from family physicians, social class, education, gender, marital status and family composition. RESULTS: The results tend to confirm that lower social class and lower educational achievement are linked to continuing smoking status. However, users of nicotine gum and patches were found to be less likely to have given up smoking than non-users. This may be explained by the fact that only those who find it difficult to give up will go to the time and expense of considering such aids. These findings could also be explained by the fact that gum and patches have only been available recently in England. The pattern of results also suggests that 'willpower' is a marker for success in cessation as opposed to a determinant of giving up in its own right. CONCLUSION: This research confirms a lot of other research on the determinants of smoking cessation and has for the first time looked at the use and impact of nicotine replacement therapy on giving up in a representative English adult population. More research is needed on observational data sets in order to tease out the true impact of nicotine replacement on cessation rates in representative smoking populations. PMID- 11420815 TI - Tobacco use by early adolescents in Norway. AB - BACKGROUND: This study examined trends in the prevalence of smoking and snuff use among Norwegian adolescents in lower secondary school (ages 13-15 years) from a national survey conducted every 5 years between 1975 and 1995. METHODS: Pupils completed a brief written questionnaire on tobacco use and related information. Local school administrators coordinated data collection and forwarded a sample of completed questionnaires to the National Council on Tobacco and Health for compilation and analysis. Effects for age, gender and survey year were examined using logistic regression analysis. Overall, 24,127 pupils in five different cohorts were included. RESULTS: Smoking was highest in 1975 when 45.5% of youth reported smoking either daily or occasionally (figures adjusted for age and gender). Smoking declined each year thereafter through to 1990 (23.6%) but increased in 1995 (26.0%), primarily due to an increase in occasional smoking. Like smoking, snuff use declined between 1985 and 1990 but increased in 1995. All changes across survey years were statistically significant. Smoking was higher among girls than boys, while snuff use was much higher among boys. CONCLUSION: Several hypotheses that might account for the recent increase were examined with the most likely factor being Norway's low allocation of resources for educational interventions and public information campaigns during the years studied. However, in the past 2 years Norway has undertaken a number of new tobacco control initiatives that may result in reversal of the most recent trends. PMID- 11420816 TI - The widening social class gap of preventive health behaviours in Spain. AB - As a way of forecasting future evolution of inequalities in disease burden and mortality, trends in preventive health behaviours by social class were examined. Using the Spanish National Health Survey from 1987 to 1997 the evolution of the Health Practices Index (smoking, alcohol consumption, physical exercise, sleep hours and the Quetelet index) is described for the Spanish non-institutionalised adult population. In the last 10 years the disadvantaged population of Spain (social class groups IV and V) has not opted for healthier behavioural choices. On the contrary the Health Practices Index figures have worsened during the study period. Conversely, the more affluent groups (social class groups I, II and III) have exhibited a net gain in the index. Due to this, an increase in the existing gap in health inequalities in Spain in terms of morbidity and mortality can be forecast for the near future. As time trends of a summary indicator of preventive health behaviours by social class have not been described in other countries, a replication of this study in different cultural, social and economic milieus could produce valuable information. PMID- 11420817 TI - Prenatal screening and genetics. AB - Although the term 'genetic screening' has been used for decades, this paper discusses how, in its most precise meaning, genetic screening has not yet been widely introduced. 'Prenatal screening' is often confused with 'genetic screening'. As we show, these terms have different meanings, and we examine definitions of the relevant concepts in order to illustrate this point. The concepts are i) prenatal, ii) genetic screening, iii) screening, scanning and testing, iv) maternal and foetal tests, v) test techniques and vi) genetic conditions. So far, prenatal screening has little connection with precisely defined genetics. There are benefits but also disadvantages in overstating current links between them in the term genetic screening. Policy making and professional and public understandings about screening could be clarified if the distinct meanings of prenatal screening and genetic screening were more precisely observed. PMID- 11420818 TI - Health economics. A bridge over troubled water. PMID- 11420819 TI - Alternative functions for TRAIL receptors in eosinophils and neutrophils. AB - BACKGROUND: Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in many tumour cells but only rarely in normal cells. The receptors of TRAIL belong to the superfamily of tumour necrosis factor (TNF)/nerve growth factor (NGF) receptors. Here we investigate TRAIL receptor expression and function in eosinophils and neutrophils. METHODS: Granulocytes were isolated from human blood and purified using standard protocols. Receptor expression was analysed by reverse transcription (RT)-PCR and flow cytometry. Cell death was analysed by the ethidium bromide exclusion test. Apoptosis was determined by analysing phosphatidyl serine (PS) surface exposure and morphological evaluation. RESULTS: Freshly purified eosinophils and neutrophils expressed TRAIL-R1, TRAIL-R3, and TRAIL-R4, but not TRAIL-R2 surface proteins. Stimulation of eosinophils with TRAIL resulted in either inhibition of apoptosis or no effect, depending on the individual from whom the cells were isolated. In neutrophils, TRAIL stimulation did not influence apoptosis. In eosinophils and neutrophils which showed no effect on TRAIL stimulation alone, TRAIL partially blocked cytokine-mediated antiapoptosis. CONCLUSIONS: Both eosinophils and neutrophils express functional TRAIL receptors on their surface. In contrast to tumour cells, TRAIL does not induce apoptosis in granulocytes but rather induces survival in eosinophils from approximately 50% of the donors. Alternatively, TRAIL may limit cytokine-mediated antiapoptosis under certain inflammatory conditions. PMID- 11420820 TI - Improved results after lung transplantation--analysis of factors. AB - Better recipient selection, sophisticated postoperative surveillance and new immunosuppressive and anti-infective regimens can improve the results of lung transplantation. We compared the results of lung transplants performed between 1992 and 1996 (early period; 47) and between 1997 and 2000 (recent period; 46) in a cohort study to assess which factors influenced survival. Estimates of relative hazards were adjusted for possible confounding effects with the use of Cox regression analysis. Overall 2-year survival was 70%. Survival by this time was significantly better in the recent period (82% vs. 60%; p = 0.0093). Acute rejection episodes and death due to BOS were less frequent in the recent period. There were no technical failures, and the cumulative incidence of BOS was low (34% at 5 years). The beneficial effect of the transplantation date 1997 or later at a hazard ratio of 0.33 (95% CI, 0.13-0.84) was materially changed only by the adjustment for ganciclovir prophylaxis (0.50; 95% CI, 0.09-2.91) and immunosuppression with mycophenolate mofetil (0.80; 95% CI, 0.27-2.36). After adjustment for both ganciclovir and mycophenolate mofetil, the beneficial time period effect was completely removed (1.24; 95% CI, 0.14-11.39). Immunosuppressive therapy with mycophenolate mofetil and use of ganciclovir prophylaxis in addition to careful postoperative surveillance and surgical expertise can lead to improved results after lung transplantation. PMID- 11420821 TI - No benefit from post-caesarean wound drainage. AB - AIM OF THE STUDY: A prospective randomized controlled trial to determine the benefit of caesarean wound drainage in 305 low-risk pregnant women. METHODS: Pregnant women at low risk of haemorrhage undergoing caesarean section in the Department of Obstetrics, University Hospital, Zurich, between June 1998 and July 1999 were randomised after informed consent into a no-suction group (n = 154) without post-caesarean wound drainage versus a control group with wound drainage (subfascial and subcutaneous) (n = 151). Outcome measures were perioperative decrease in haemoglobin (Hb), postpartum fever (> 38.5 degrees C for > 2 days), sonographic haematoma and other complications requiring revision, cumulative opiate dose adjusted to body weight, length of hospitalisation and operation time. RESULTS: 305 patients completed the study. Decrease in Hb and the rates of fever, haematoma and revision were similar in both groups. However, cumulative opiate dose was lower in the no-suction group (4.5 +/- 1.8 vs 2.8 +/- 1.4 injections, p = 0.0001), and hospital stay was shorter (6.5 +/- 2.4 vs 7.4 +/- 2.8 days, p = 0.0058), as was operation time (32.7 +/- 11.3 v 36.1 +/- 10.5 min; p = 0.0071). CONCLUSIONS: Routine post-caesarean wound drainage is not only useless but cost-ineffective. In the light of our results, wound drainage may be questioned and should be analysed generally. PMID- 11420822 TI - Use of the Internet by medical doctors in Switzerland. AB - QUESTIONS UNDER STUDY: To investigate the utilisation of the Internet by primary care physicians for medical purposes during their daily practice, and to clarify the reasons for use or non-use of this technology. METHODS: Cross-sectional postal survey in German-speaking Switzerland employing a purpose-designed pre validated 69-item questionnaire. A random sample of 2009 primary care physicians participated in the investigation. MAIN OUTCOME MEASURES: Number of primary care physicians with access to the Internet; reasons for using the Internet during consultations; sources of information in solving medical problems arising from concurrent patient care. RESULTS: 55% of the physicians returned the completed questionnaire. 75% of respondents reported access to the Internet. Only 7% use the Internet during patient consultations. The main reasons for not using the Internet were time pressure and concerns about potential negative interaction with physician-patient communication. To solve patient-specific problems arising during daily practice, 59% of the practitioners consult text-books or colleagues. Only 14% of respondents report regularly finding useful information on the Internet. Internet users assess information quality by checking on authorship, institution, publishing company, or whether the information is sponsored by a third party with a potential conflict of interest. CONCLUSIONS: Access to the Internet is widespread amongst German-speaking Swiss primary care physicians. Only a small minority use the Internet for information retrieval during consultation hours. Electronic information systems need to be tailored to the needs of primary care physicians. PMID- 11420823 TI - [Only 2 glasses of beer per day. Heart is healthy, liver destroyed?]. PMID- 11420824 TI - [Initial success with topical immunomodulators. A creme for skin cancer? (interview by Renate Jackle)]. PMID- 11420825 TI - [Treatment of scarlet fever with homeopathy? MMW readers remember the pre penicillin era]. PMID- 11420826 TI - [Tinnitus: investigating etiology, then control. Water sounds instead of ear whistling]. PMID- 11420827 TI - [An asthmatic patient with concomitant heart disease. Can beta mimetics be administered?]. PMID- 11420828 TI - [Long-term weight reduction. Current therapy approaches in obesity]. AB - A large percentage of obese patients can lose weight with a combination of various dietary measures. The following possibilities are available: 1. The fat reduced, carbohydrate-enhanced diet. 2. Formula diets, which are applied when a mixed diet has failed, or, alternatively, as an "introduction" to a weight reducing program. In addition, suitable exercise programs and behavior-modifying measures aimed at stabilizing weight losses over the long-term, are also recommended. If this basic program fails, adjunctive drug treatment is indicated. Surgical treatment may be considered when various conservative forms of treatment have proved unsuccessful over a period of years. PMID- 11420829 TI - [Insulin therapy in the type 2 obese diabetic patient. Supplementing the deficit]. AB - The aim of insulin therapy in obese type 2 diabetics is to achieve a near-normal glycemic serum sugar metabolism while avoiding any further increase in weight and hypoglycemia. A further aim is to achieve maximum flexibility in the patient's lifestyle. For this purpose, in addition to bedtime administration of HPH insulin aimed at achieving a morning blood sugar level of < 100 mg/dl, regular insulin or a rapid-acting insulin analog should be administered at mealtimes. Administration of NPH insulin during the day is not necessary in most overweight type 2 diabetics--since the residual capacity for endogenous insulin secretion is adequate--and should be applied only when a requirement has been confirmed (basal rate test). Through the additive administration of oral (metformin, acarbose), metabolic control can be improved and the insulin dose simultaneously reduced, with associated positive effects on the patient's weight. With the aim of lowering the roughly three-fold increase in cardiovascular mortality rate in type 2 diabetics, optimal antihyperglycemic control should be accompanied by optimal management of blood lipids (LDL < 100 mg/dl, HDL > 45 mg/dl, triglycerides < 150 mg/dl), and blood pressure (RR < 135/80 mm Hg). PMID- 11420830 TI - [The new oral antidiabetics. Type 2 diabetes under control only with tablets?]. AB - The new oral antidiabetics have expanded our possibilities of treating type 2 diabetes and associated diseases. AGIs and insulin sensitizers represent new principles of action. The insulinotropic substances improve the dynamics of insulin secretion. They enable pathophysiologically-oriented treatment that takes account of the patient's need for simple, low-stress management. This improves the quality of life, in particular of the elderly and disabled patient. However, this fact does not justify delaying treatment with insulin, which is always needed when the goals of treatment cannot be achieved with oral preparations. PMID- 11420831 TI - [Emergencies in general practice, 16: Transitory ischemic attacks. When suspected ASS and heparin are contraindicated!]. PMID- 11420832 TI - [Gas station attendant with chronic recurrent abdominal pain. Unusual lead exposure lead to secondary porphyria]. PMID- 11420833 TI - [The wound. 3: Specific treatment principles]. PMID- 11420834 TI - [Sudden onset in the long distance driver--"strange protrusion" of the upper arm. Rupture of the tendon of the long biceps brachii muscle head]. PMID- 11420835 TI - [Heart and lung disease. Are beta blockers allowed in asthma?]. PMID- 11420836 TI - [Decubitus ulcer. Biological wound treatment stimulates the healing process]. PMID- 11420837 TI - [Chronic polyarthritis]. PMID- 11420838 TI - [Cardiovascular prevention by moderate alcohol drinking? Not every heart benefits!]. PMID- 11420839 TI - [Animal models of psychiatric diseases: possibilities, limitations, examples and demonstration of use]. AB - Animal models of psychopathology serve as a central tool for psychopharmacologists in their attempts to develop new, more efficient medications for psychiatric disorders, and in the efforts to explore the mechanisms of conventional and novel drugs. The development of efficient models for psychiatric diseases is complicated since the mechanisms of the disorders are not clear, major parts of the diagnosis depend on verbal communication with the patient and many of the symptoms are expressed mainly through the subjective experiences of the afflicted individual. Inspite of these difficulties, effective models were developed for most psychiatric diseases. The development of such models is based on their validation in three different dimensions: face validity- behavioral similarities between the model and the disorder; construct validity- similarities between the mechanisms related to the model and a mechanistic theory of the disease; predictive validity--that the model response to the conventional medications that are effective in the disease and will not respond to drugs that are not effective in the disease. The present paper presents three known models of depression; each induced in a different way and therefore represents a group of models: Reserpine-induced hypoactivity--represents the group of pharmacologically induced models. Forced swim test--represents the group of behaviorally induced models. Flinders Sensitive Line--represents the group of genetically induced models. The use of these models in an attempt to examine the range of action of a new potential antidepressant and its mechanisms of action is demonstrated with a recent set of experiments with inositol. PMID- 11420840 TI - [Laparoscopy as part of the evaluation and management of ovarian and cervix neoplasms]. AB - INTRODUCTION: Surgery is the treatment of choice in most early stages of cervical cancer and advanced stages of ovarian cancer. Failing to preoperatively diagnose para-aortic and parametrial metastases in cervical cancer or a non-resectable, ovarian cancer, may results in a superfluous laparotomy. AIM: To evaluate the advantage of using laparoscopy in cervical and ovarian cancer. PATIENTS AND METHODS: Study population includes patients with ovarian or cervical cancer referred between 1997-1999. A CT scan and a trans-vaginal sonography were used to detect involvement of pelvic and para-aortic lymph nodes, parametrium and other metastases. In patients with cervical cancer, para-aortic lymph node dissection was laparoscopically performed. When para-aortic nodes were negative and parametrium was clear, radical hysterectomy and pelvic lymph nodes dissection was conducted through laparotomy. When para-aortic lymph nodes or parametrium were positive, patients were referred for radiation therapy. In ovarian cancer patients, the upper abdomen and the pelvis were examined laparoscopically to evaluate the possibility of optimal debulking surgery. Staging was done for patients who were not candidates for optimal debulking surgery and second debulking surgery was considered. RESULTS: Nine patients with stage 1b-11a cervical cancer were included. Preoperatively, 2 of them were suspected for lymph node involvement. Following laparoscopy one was confirmed to have para-aortic lymph node involvement and the other did not. In the remaining 8 patients, one was found to have parametrial involvement and laparotomy was avoided, while the rest were treated surgically. Eighteen ovarian cancer patients were included in this study. Laparoscopy revealed an extensive disease in 7 patients and therefore staging laparoscopy was completed. In the remaining 11 patients, laparotomy was performed, operable disease was found and complete debulking surgery was conducted. In only one patient of the 11 complete debulking surgeries was not possible. CONCLUSIONS: In accordance with the experience and skills of the surgical team, we propose utilizing laparoscopy in cases where laparotomy may be avoided. PMID- 11420841 TI - [Sentinel lymph node biopsy in breast cancer: a validation study and preliminary results]. AB - INTRODUCTION: Sentinel lymph node biopsy (SLNB) has been recently proven to be an accurate staging method for breast cancer, replacing axillary lymph node dissection (ALND) in selected cases. We present our initial experience and the process of introduction and implementation of SLNB in a University Hospital setting. MATERIAL AND METHODS: 46 SLNB were performed in 42 consecutive female patients with invasive breast cancer. Treatment included 0.4mCi-2mCi of Tc-99m rhenium colloid injected either 2 hours before surgery (0.4 mCi) or the night before surgery (2 mCi). Four milliliters of Patent Blue V were injected peritumoral 10 minutes prior to skin incision in all patients. Following SLNB all women underwent subsequent ALND. Sentinel nodes were processed both with multiple (10-15) H&E sections and immunohistochemistry with cytokeratin antibodies stain. RESULTS: Blue dye, isotope or the combination of both identified 43/46 (93%) of the sentinel lymph nodes. ALND was performed only unilaterally in 4 patients with bilateral breast cancer bringing the total evaluable SLNB to 39. In the 39 patients in whom the sentinel node was successfully identified and underwent ALND, the SLNB was true positive (TP) in 17/39 (44%) true negative (TN) in 20/39 (51%) and false negative in 2/39 [(5%), both T2 lesions] with overall accuracy of 95%. In the last 10 cases all sentinel nodes were successfully identified with 70% TP and 30% TN. CONCLUSIONS: Experience with at least 30-40 consecutive cases for safe implementation of SLNB in clinical practice. Specific training and dedication is required for the entire team involved, including surgeons, nuclear medicine physicians and technicians and pathologists. PMID- 11420842 TI - [Eosinophilic ascites as a presenting symptom of the hypereosinophilic syndrome]. AB - We report a case of a 22 year old man with a history of bronchial asthma, suffering from diarrhea, eosinophilic ascites and prominent blood and bone marrow eosinophilia. The patient responded to corticosteroid treatment. Later, hydroxyurea was added to this treatment because of recurrence of eosinophilia, ascites and hepatosplenomegaly. Eosinophilic ascites is rarely the outstanding symptom of the hypereosinophilic syndrome. Following the exclusion of other reasons for eosinophilia, concomitant unexplained hepatosplenomegaly suggested the diagnosis of a hyperereosinophilic syndrome. PMID- 11420843 TI - ["Stealing madness"--kleptomania]. AB - Kleptomania is one of the common less diagnosed impulse control disorders, which could be treated by the combination of psychological and pharmacological therapy. The most effective treatment regimens include cognitive behavioral therapy and interpersonal psychotherapy in the psychological field, and antidepressants and mood stabilizers as pharmacological treatment. Most of the patients with kleptomania are initially treated only for a comorbid psychiatric disorder, since kleptomaniac symptoms had not been raised in the anamnesis. The aim of our article is to inform the physicians about the possible diagnosis and treatment options for this disorder in order to prevent co-morbidity with other psychiatric disorders. Some specific cases are presented in order to explain symptomatology. PMID- 11420844 TI - [The lack of awareness of the Israeli population regarding gastrointestinal complications from non-steroidal anti-inflammatory drugs]. AB - Non-steroidal anti-inflammatory drugs (NSAID) are potent analgesic agents. They are indicated for rheumatoid arthritis, osteoarthritis, severe acute and prolonged chronic pain. A wide range of analgesic agents are available. The objective of this study was to assess the patient's knowledge of his illness, his knowledge familiarity with NSAID medications and their side effects and thereupon, to evaluate the awareness, utilization, satisfaction and expectations associated with current available treatment. During the year 2000, a randomized telephone screening questionnaire, was completed over a 2 month period for a sample group of the general Israeli population over 50 years of age. Of the 3179 persons contacted, 2028 responded and 292 persons were treated with NSAIDs and further investigated. Fifty percent suffered from chronic musculoskeletal pain, 22% had osteoarthritis, 10% had rheumatoid arthritis, and 18% had undefined chronic pain. The average age of the respondent was 65 years old. The average duration of treatment was 11 years. The most common drugs used by the 292 respondents that were taking NSAIDs were diclofenac (Voltaren)--51% and naproxen (Naxyn)--21%. Only 4% were treated with COX-2 specific inhibitors. Twenty percent of the respondents utilized more than one NSAID simultaneously. Forty seven percent of the patients had peptic ulcer disease but only 39% of them were simultaneously treated with an NSAID and a gastroprotective agent. Sixty eight percent of patients expected amelioration of symptoms while taking NSAIDs. Seventy seven percent of respondents were dissatisfied with treatment. The patients expressed that they did not have a reliable easily accessible source of information on their disorder or on available treatment options. Many respondents were not aware of their medical diagnosis. In general, patients were not aware of all the available NSAIDs or the side effects of gastrointestinal complications. Enhancement of information available to the patient is warranted to improve the provision of efficient therapy, reduce side effects and increase patient satisfaction from therapy. PMID- 11420845 TI - [Aggressive surgical treatment for cholangiocarcinoma]. AB - Surgery remains the only curative approach to cholangiocarcinoma. Despite operative difficulties concerning tumor location, vascular involvement or hepatic invasion curative resection (i.e. achievement of tumor free margins) carries good survival rates and is associated with low morbidity and mortality. Our experience with 31 patients with cholangiocarcinoma operated during the years 1993-1999 is presented. Twenty-five patients were found to have hilar cholangiocarcinoma, 3 carcinoma of mid-choledochus and 3 distal cholangiocarcinoma. All patients were evaluated by a diagnostic protocol including laparoscopy and laparoscopic ultrasound. Surgery consisted of local resection of the tumor with Roux en Y hepaticojejunostomy reconstruction. Three patients had concomitant hepatic resection for hilar cholangiocarcinoma while 3 patients had pancreaticoduodenectomy for distal cholangiocarcinoma. Adenocarcinoma was diagnosed in all patients. Curative resection was achieved in 19 patients and palliative resection (microscopically involved resected margins) in 12. One patient died at 60 days postop (3.5% mortality). Postoperative complications included cholangitis occurring in 10 patients who were referred with biliary drains, intraabdominal abscess (2 patients) and biliary leakage (1 patient) all treated conservatively. Overall survival rate was 82% at one year, 73% at two years and 45% at 5 years. When comparing curative resection with palliative resection, survival rates were 92% versus 71% for 1 year, 72% versus 50% for 2 year and 57% versus 14% for 5 years. Selection of patients using Laparoscopic Ultrasound combined with aggressive surgical approach enable curative or palliative resection 15 the only treatment modality which is significantly improved survival. PMID- 11420846 TI - [Treatment of traumatic false aneurysm of the thoracic aorta with stent graft]. AB - The treatment of traumatic false aneurysm of the thoracic aorta by endovascular stent graft may have advantages over conventional surgery. This is a case study of two women suffering from false aneurysm of the thoracic aorta caused in one by a knife injury and in the other by a car accident. After the patients became hemodynamically stable, a commercially available endothelial stent graft (Talent, Gor) was deployed. Recovery was rapid in the first patient. The second patient required emergency laparotomy for venous bleeding one day after stent placement; she died two weeks later, mainly from organ failure. CONCLUSIONS: Endovascular techniques can be used in selected cases to treat thoracic false aneurysms thereby avoiding the complexity and morbidity of conventional surgery. PMID- 11420847 TI - [Clinical mentorship as a bridge to life in the treatment of eating disorders]. AB - The aim of this article is to describe a community-based multi-modality intensive treatment program, which demonstrates a significant reduction in symptoms of eating disorders while providing an opportunity to deal with the functional and social skills. Treatment is delivered through a multidisciplinary team. The uniqueness of the team is in the inclusion of clinical mentors. These are social workers, art therapists and graduate level psychology students who are trained to connect with clients in an informal, intensive manner (10-40 hours a week). The mentors focus on legitimizing healthy attitudes to food, eating and life, while highlighting the pain and loss associated with the disorder. The mentors serve as meal companion and calming figure, representing the healthy self-caring voice. The relationship established during the many hours spent together becomes a powerful experience, completely different from the isolation of the disorder. The treatment intervenes in most areas of life, focusing on an active search for possibilities, in contrast to the emptiness associated with the disorder. This article presents the results and principles of a 2.5-year practice. Seventeen patients, ill for 6 years and more, completed this program more than a year ago. An assessment of their BMI and their general outcome using Eckert scales was performed. One year after completing the program, 76% of the patients were defined as recovered and 12% were almost recovered with only a few remaining symptoms. All of these patients function satisfactorily in the community, both in social and occupational aspects. Six percent were partially recovered and 6% suffered from regression during the first year of follow-up. This treatment provides the intensity required in hospitalizations, while enabling patients to stay in the community and maintain those activities that survive the disorder. This program has proven to work well with chronic patients. Further data, not analyzed as yet, also indicates the efficacy of this treatment in acute cases. PMID- 11420848 TI - [Following 90 patients with antiphospholipid syndrome with antibody titers and correlations with clinical manifestations: symptoms of the disease, a new antibody and correlations with clinical manifestations in the Israeli population]. AB - The antiphospholipid syndrome (APS) is defined by the presence of antiphospholipid antibodies (aPLs) with venous or arterial thrombosis, recurrent pregnancy loss or thrombocytopenia. The syndrome can be either primary or secondary to an underlying condition, most commonly systemic lupus erythematosus (SLE). The purpose of the study was to review the manifestations of the APS in Israeli patients and to investigate the difference between patients with primary or secondary syndrome. Analysis of the correlation between the manifestations of the disease and the antibody titers [LA (lupus anticoagulant), IgG aCL (anticardiolipin), IgM aCL] was also performed. In the study there were 90 patients with APS in the Sheba Medical Center. The clinical findings for all patients were recorded according to established protocol. RESULTS: Arterial thrombotic events were present in 51.1% of the patients; cerebral ischemic attacks were the most frequent arterial events. Venous thrombotic events were present in 45.6% patients; deep venous thrombosis was the most frequent venous event. Obstetric complications were found in 37.3% of patients; the most frequent complication was abortions. The clinical findings of patients with primary or secondary syndrome were similar. Patients with APS associated with lupus had a high prevalence of hemolytic anemia (28.6% v 3.3%; P = 0.001), and ANA (antinuclear antibody) (75% v 12.9%; P = 0.0001). There was no correlation between the antibody titers (LA, IgG aCL, IgM aCL) and clinical manifestations. The patients received anticoagulant treatment and antiaggregant drugs to prevent recurrence of symptoms. Clinical manifestations in Israeli patients did not differ from those in non-Israeli patients. CONCLUSIONS: Venous or arterial thrombosis and obstetric complications are the most frequent clinical findings in APS. There was no different in clinical manifestations between primary and secondary syndrome. The findings of autoimmune hemolytic anemia and ANA were more frequent in patients with secondary APS. There was no correlation between antiphospholipid antibody titers and manifestations of disease. Overall, the patients require therapy with anticoagulant treatment and antiaggregant drugs. The clinical manifestations were similar in Israeli and non-Israeli patients. PMID- 11420849 TI - [Surgery of the carotid body tumors]. AB - Carotid body tumor is a rare tumor of obscure origin, usually benign, and commonly present as a non-painful cervical mass. Resection is the treatment of choice. When the tumor is very adherent to the carotid artery, it may be necessary to also resect the artery and to reconstruct the arterial continuity using a saphenous or synthetic interposition graft. The most common complication of surgery is damage to the cranial nerves in the vicinity of the artery. We present 8 patients with carotid body tumor who underwent surgery in our department in 1996-1999. Two had a large tumor penetrating the intima which mandated resection of the involved carotid artery and a saphenous interposition graft reconstruction. Our experience with resection of carotid body tumors is detailed, with emphasis on precautions taken to prevent damage to the adjacent cranial nerves. PMID- 11420850 TI - [The unexpected features of the vaccine against chickenpox]. PMID- 11420851 TI - [Fasting before surgery: is it time to slaughter the sacred cow?]. PMID- 11420852 TI - [Idiopathic hypereosinophilic syndrome--update]. PMID- 11420853 TI - [Intra-articular injections of hyaluronic acid in the treatment of knee osteoarthritis]. PMID- 11420854 TI - [Leptin--pathology and physiology]. PMID- 11420855 TI - [Sepsis associated cholestasis in adults]. AB - Hepatic injury presenting as jaundice, hypglycemia, encephalopathy, coagulation disturbances and a significant increase in transaminases in a late well recognized complication of sepsis. Less known is a cholestatic liver injury, which can precede sepsis. This type of early liver injury is characterized by a rise of direct bilirubin and less prominent increase in liver enzymes. Experimental work in animal models demonstrates that sepsis associated cholestasis is mediated by a variety of gram negative bacterial endotoxins and cytokines which interfere with the function of bile acid intracellular transporters. Although sepsis associated cholestasis is quite common, its clinical and prognostic characteristics are not widely appreciated. This often results in performance of unnecessary procedures and delayed diagnosis. Here we review the relevant updated literature regarding the pathophysiological basis of this phenomenon and its clinical presentation and implications. PMID- 11420856 TI - [Thalidomide and cancer--revival of a drug]. PMID- 11420857 TI - [Pericardial involvement in scleroderma]. PMID- 11420858 TI - [The ultrasound markers of the aneuploid fetus in the first and second trimesters of pregnancy]. PMID- 11420859 TI - [The revival of an old diagnostic marker: C-reactive protein]. PMID- 11420860 TI - [The Swan-Ganz catheter--past, present and future]. PMID- 11420861 TI - [The representation of the egocentric space in the posterior parietal cortex]. PMID- 11420862 TI - [Contraception during lactation]. PMID- 11420863 TI - [A star is born]. PMID- 11420864 TI - The longitudinal case study: from Schon's model to self-directed learning. AB - BACKGROUND: Rapid changes observed in information technologies, medical practice, and learning methods encourage physicians to develop new updating strategies. To test its feasibility and to help physicians devise new learning and updating strategies, the knowing-in-action model developed by Schon was applied in planning and evaluating an interactive workshop. Acquisition of knowledge was tested. METHODS: The office and hospital charts of a family physician were reviewed. They were used to prepare a longitudinal case study, based on the real life story of a hypertensive patient followed by her doctor over a period of 15 years. The clinician's approach to solving clinical problems was triangulated for credibility with general practitioners, specialists, and the information available in the literature. This longitudinal case study was used to develop an interactive educational workshop. The workshop was presented to physicians who had registered in an accredited continuing medical education event. Changes in pre- and postevent knowledge among the participants were assessed using touch pad technology to evaluate the effectiveness of this approach on the acquisition of knowledge related to management of arterial hypertension and associated clinical problems. RESULTS: A comparison of pre- and post-test data showed a significant improvement in knowledge for participants who answered all questions on both questionnaires (n = 8/37). The average score of these participants increased from 5.5 of 10 before the workshop to 8.3 of 10 after the workshop (p < .05). Participants reported a high satisfaction rate for the event. FINDINGS: A workshop using the longitudinal case study enables physicians to perceive their daily practice through a continuing education activity in which they experience the processes of reflection in action and reflection on action described by Schon. It also increases awareness of the gap between current practice and experts' recommendations and provides an opportunity to evaluate the means for bridging or closing this gap. It sensitizes the physician to patients' changing needs and prompts the clinician to reflect on the who, what, when, where, and how of learning. PMID- 11420865 TI - A continuing medical education lecture and workshop, physician behavior, and barriers to change. AB - BACKGROUND: Continuing medical education (CME) is undertaken with the intention that it will affect the practice of medicine at the level of choices made by individual physicians. Inherent in this effort is the assumption that CME is sufficient to effect a change in physician behavior. METHODS: To further examine the relationship between a CME activity and physician behavior, we conducted a study of behavior and barriers to change associated with a CME lecture and workshop on breast cancer risk assessment and treatment. Using the assessment of learning outcomes model of the International Association of Continuing Education and Training, we developed an instrument for assessing physician behavior and barriers to change. RESULTS: Throughout the United States and Canada, the instrument was administered on-site immediately after a CME activity implemented at 79 hospitals and cancer centers. It was administered again 6 months after the CME activity. There were 1,244 responses collected from 4,537 participants. This study reports the survey findings of 176 physician-paired responses to both the first and second waves of surveys. Some physicians changed their behavior with regard to performing risk assessments on all of their eligible patients. Ninety two of the 176 physicians indicated that they had changed their practice regarding the use of tamoxifen therapy. Twenty-one physicians indicated that they were already using tamoxifen in their practice setting. Three influential barriers to change were identified: a lack of consensus among colleagues and peers, lack of time for assessment and patient counseling, and lack of reimbursement by the patient's insurance companies. FINDINGS: The CME activity was effective in changing the self-reported behavior of some physicians. Others attended the CME activity to obtain more information or to become more skilled about a procedure they had already implemented. Because of formidable barriers, it is unlikely that a single educational intervention will be sufficient to effect a change in the clinical practices of all physicians who participate in a CME activity. PMID- 11420866 TI - Tolerance of ambiguity among MD/MBA students: implications for management potential. AB - BACKGROUND: Health care is increasingly characterized by uncertainty and turbulence. In an environment of rapid change, flexibility is critical to the success of managers and organizations. Future physician executives must also be open to change and must be able to deal with the uncertainties of management; they must be able to tolerate the ambiguity in management situations. METHOD: This study uses tolerance of ambiguity measures to analyze students at six medical schools offering dual-degree (MD/MBA) programs. Students enrolled in dual degree programs were assessed and compared with a control group of traditional medical students. RESULTS: MD/MBA students exhibit a higher tolerance of ambiguity than traditional medical students. FINDINGS: As a characteristic associated with leadership ability, tolerance of ambiguity offers a potential indicator of future success as a physician executive. As such, tolerance of ambiguity might be used for selective admissions to medical school and as an indicator of a student's potential to transition between clinical and management functions. As students match personality traits with career choices, those who serve their learning needs must anticipate differences across selected disciplines, roles, and responsibilities. PMID- 11420867 TI - Learning, evaluation, and the tao of continuing medical education. PMID- 11420868 TI - Reconsidering "good teaching" across the continuum of medical education. AB - There is no shortage of sustained inquiry into the nature and evaluation of teaching in medical education. For the most part, however, this growing and respectable body of inquiry has uncritically adopted a single model of effective teaching that is assumed to be appropriate across variations in context, learners, and teachers. This article presents five alternative views of "good teaching" and challenges the trend toward any single, dominant view of what constitutes good teaching. Based on 10 years of research, in five different countries, studying hundreds of educators in adult and higher education across a wide range of disciplines, contexts, and cultures, we have evidence of five different perspectives on good teaching: transmission, developmental, apprenticeship, nurturing, and social reform. Each perspective represents a philosophical orientation to knowledge, learning, and the role and responsibility of being an educator. A "snapshot" of each perspective is provided, including an example from continuing medical education (CME), a set of key beliefs, primary responsibilities, typical strategies, and common difficulties. Readers are encouraged to use the five perspectives as a means of identifying, articulating, and revisiting assumptions and beliefs they hold regarding their view of effective teaching. They are also encouraged to resist a "one-size-fits-all" approach to the investigation, improvement, or evaluation of teaching in CME. PMID- 11420869 TI - Commitment to change statements: a way of understanding how participants use information and skills taught in an educational session. AB - BACKGROUND: Commitment to change has gained increasing use in assessing short course effectiveness. This study examined the changes that learners intended to make in practice following an intensive day-long course offered at multiple sites, counted changes relative to the curriculum's focus, and analyzed which changes were implemented in practice. METHODS: Participants at a course on the management of male sexual dysfunction were asked to identify the changes to which they would commit. Six months after the course, they were asked to indicate which changes they implemented fully, partially, or not at all. RESULTS: A total of 352 physicians attended the courses held in 21 centers. A majority of attendees (344 or 97.7%) completed forms at the end of the course, providing 1,635 commitment statements. Six months later, 197 (57.3%) physicians provided follow-up data about 935 (55.4%) of the commitment statements originally submitted. Of these, 602 (66.52%) were completely implemented. Many of the changes related to two specific aspects of the course, namely, sexual history taking and medical intervention, accounting for 45.93% of the intended commitments and 47.67% of the changes completely implemented. Slightly over half (58%) of the course time was devoted to these two areas. There was a significant correlation between the number of changes and the amount of time allocated to that content within the course. FINDINGS: Commitment to change statements offered by course participants can be used to examine the impact of a course relative to its learning focus. Continuing medical education providers must take a critical look at commitment to change statements as an "intervention" in their own right and determine how the tool can best be used as a continuing medical education intervention. PMID- 11420870 TI - Value of unstructured time (breaks) during formal continuing medical education events. AB - BACKGROUND: Unstructured time (breaks) at formal continuing medical education (CME) events is nonaccredited in some jurisdictions. Program participants, however, perceive this time as valuable to their learning. The purpose of this research was to determine what activities occur during unstructured time in formal CME events and how these activities impact learning for physicians. METHODS: A qualitative method based on grounded theory was used to determine themes of behavior. Both individual and focus group interviews were conducted. Data were analyzed and coded into themes, which were then further explored and validated by the use of a questionnaire survey. RESULTS: One hundred ninety-seven family physicians were involved in the study. Several activities related to the enhancement of learning were identified and grouped into themes. There were few differences in the ranking of importance between the themes identified, nor were differences determined based on gender or type of CME in which the break occurred. FINDINGS: The results suggest that unstructured time (breaks) should be included in formal CME events to help physician learners integrate new material, solve individual practice problems, and make new meaning out of their experience. The interaction between colleagues that occurs as a result of the provision of breaks is perceived as crucial in aiding the process of applying knowledge to practice. PMID- 11420871 TI - Evaluation of the effectiveness of an international diploma course in tropical medicine. AB - BACKGROUND: Numerous impediments to conducting continuing education (CE) courses in remote sites, particularly those courses that take place in developing countries, can include challenges associated with planning, infrastructure, and financial risk. This study reports the effectiveness of a course planned in the United States and executed in Peru, the Gorgas Course in clinical tropical medicine. METHODS: A survey was conducted of participants who had completed the Gorgas Course as recently as 6 months and as long as 3 years earlier. The questionnaire sought to determine each participant's reason for participation, whether the course was instrumental in the participant's reaching the personal goal associated with participation, and whether the participant considered the course to be worth the time and money spent to enable participation. RESULTS: Forty-nine participants responded to the questionnaire, all of whom indicated that the Gorgas Course enabled achievement of the personal goal associated with participation. Fully 100% of course participants stated that participation was worth the time and monetary expenditure, most often citing their having access to patients with tropical diseases and the personal enrichment of living overseas as reasons the course was worth its high cost. FINDINGS: It is logistically and financially feasible to conduct CE courses in developing countries, provided that the organization in the planning country has strong, pre-established relationships with the host institution(s). Continued collaboration between planning partners and frequent, rigorous course evaluations are necessary to enable an international CE course to become a stable, continuous academic offering. PMID- 11420872 TI - When is it time to start dialysis? PMID- 11420873 TI - Elevated serum creatinine concentration--a marker for adverse operative outcome. PMID- 11420874 TI - Advanced glycation end products: specific fluorescence changes of pentosidine like compounds during short daily hemodialysis. AB - BACKGROUND: Advanced glycation end products (AGE) accumulate in uremia and represent an important etiopathogenetic cause of morbidity in dialyzed patients. Conventional hemodialysis treatment seems to be ineffective in lowering AGE levels. We wished to investigate whether daily hemodialysis (DHD), a treatment that seems to result in better clinical condition in end-stage renal disease patients, is effective in the reduction of these compounds. METHODS: We evaluated 10 non-diabetic patients on standard hemodialysis (SHD = 3 x 4 h/week) for more than 6 months by a crossover study. These patients were assigned randomly to 6 months of DHD (6 x 2 h/week) or 6 months of SHD. Then, they were switched to 6 months of the alternative treatment. At the end of these two periods, we studied pentosidine-like AGE compounds by measuring the total fluorescence at a wavelength characteristic for these substances: Ex: 335nm/Em:385nm; we also measured protein-linked pentosidine at the same time points. Finally, we determined the AGE-related total fluorescence in the deproteinized serum of 13 uremic patients on peritoneal dialysis (CAPD) and of 10 healthy controls. RESULTS: Pre-HD AGE-related total fluorescence obtained after 6 months of DHD was significantly lower than that obtained with standard HD (DHD = 201.3 +/- 36.4 AU/ml vs. SHD = 267.5 +/- 141.4 AU/ml, p = 0.03). The extraction rate per minute of dialysis was slightly, but not significantly higher during DHD than SHD (0.29 +/- 0.11% vs. 0.23 +/- 0.04, p = 0.07). AGE-related total fluorescence pre-HD values in patients treated by SHD and DHD were about 20-fold higher than in control subjects. They did not differ from CAPD patients. The pre-dialysis level of protein-linked pentosidine was significantly lower in DHD than in SHD (DHD = 16.12 +/- 4.71 pmol/mg protein, SHD = 22.64 +/- 6.86 pmol/mg protein, p < 0.01). CONCLUSIONS: DHD showed a reduction in AGE-related total fluorescence, although the mean value remained higher than in control subjects. DHD is also accompanied by a decrease in protein-linked pentosidine. PMID- 11420875 TI - Short- and long-term mechanical cardiac assistance. AB - With the increase in high risk patients undergoing cardiac surgery and the substantial mortality among patients waiting for cardiac transplantation, the need for mechanical circulatory support is growing. Several devices are currently available, ranging from the intra-aortic balloon pump to fully implantable ventricular assist devices. Each system has its own features, and proper patient selection as well as the timing of implantation is sometimes difficult. Algorithms for stepwise management in subgroups of patients remain controversial and the concepts of weaning patients after myocardial recovery during mechanical circulatory support need further evaluation for their long-term effects. Future identification of valuable prognostic and risk factors may help in decision making and allow for improved survival of these often very ill patients. In this report we review the concepts of mechanical circulatory support at our institution with emphasis on a detailed overview of technical features of extracorporeal life support. PMID- 11420876 TI - IABP assistance: a test bench for the analysis of its effects on ventricular energetics and hemodynamics. AB - IABP assistance is frequently used to support heart recovery, improving coronary circulation and re-establishing the balance between oxygen availability and consumption. Hemodynamic and energetic parameters (endocardial viability ratio, ventricular energetics) are used to evaluate its effectiveness which depends on internal (timing, balloon volume and position) and external factors (circulatory conditions). Considering short, medium and long-term effects of IABP, the first depends on its mechanical action, the latter on the changes induced in circulatory parameters. The analysis of the first is important because conditions for the onset of a virtuous cycle able to support ventricular recovery are created. Simulation systems could be helpful in this analysis for the implicit reliability and reproducibility of the experiments, provided that they are able to reproduce both hemodynamic phenomena and energetic relationships. The aim of this paper is to present a system originally developed to test mechanical heart assist devices and modified for IABP testing. Data reported here are obtained from in vitro experiments. A partial verification, obtained from the literature is presented. PMID- 11420877 TI - Influence of ischemic time and temperature on endothelial cell growth after transport. AB - BACKGROUND: The preparation of tissue-engineered material is a complex procedure. The possibility to transport tissue between laboratories without losing endothelial cell (EC) function was examined. METHODS: In 3 month old juvenile sheep (n = 6) a piece of vein (n = 14) was harvested and transported over 900 km to the tissue laboratory in Dulbecco's Modified Eagle's Medium (= DMEM). Vein material of each animal was transported at 4 degrees C (Group I, n = 6) and 25 degrees C (Group II, n = 8). EC growth potential was evaluated in function of the medium temperature and the ischemic time (between 8-24 hours). At the end of the first passage the EC of Group I and II were put together to save autologous serum of the sheep. After the 2nd passage the EC were cryopreserved at -80 degrees C to evaluate if EC viability would change. RESULTS: The growth potential of hypothermic Group I was equal in 16.7% (n = 1), higher in 33.3% (n = 2) and lower in 50% (n = 3) than Group II which had the same ischemic time during transport. Increase in ischemic time up to 24 hours showed no decrease of growth potential. Cryopreservation had no significant influence on EC viability. Viability at the end of the second passage, after recultivation and at the end of the third passage was 97.4% +/- 1.52, 95.5% +/- 1.34 and 94.5% +/- 1.08 respectively. CONCLUSIONS: In sheep there is no need to transport the EC at a temperature of 4 degrees C. Up to 24 hours growth potential and viability are maintained also at 25 degrees C. PMID- 11420878 TI - A mathematical model of the fetal cardiovascular system based on genetic algorithms as identification technique. AB - The development of fetal cardiac surgery, considered the ultimate goal in the treatment of congenital cardiac malformations, needs to be supported by detailed knowledge of the blood circulation in the fetal cardiovascular system. The hemodynamic behavior in distal territories is usually inferred from vessel resistance indices, which give limited physiological information. This study presents a mathematical model of the human fetal global cardiovascular system, developed to clarify the relationships and differences existing between upper and lower body circulation. We modelled the heart with two time-varying capacitances, each representing the respective ventricle's pressure-volume relationship. The fetal vascular system was represented using two six-element Windkessel models, for the upper and lower body respectively. We obtained the identification of the set of circuital and elastance function parameters of the model using Genetic Algorithms (GAs), which follow the laws of evolutionary theory. We compared the results of our numerical study on the model identified with data collected from measurements and literature, to validate the proposed global cardiovascular system model of the human fetus. This model is intended as an instrument to investigate the differences in blood distribution between the different vascular districts in the upper and lower fetal body and the role of the aortic isthmus, the small tract of vessel connecting upper and lower fetal vascular beds; it may also represent a useful tool in the assessment of dynamic balance during mechanical assistance of circulation. PMID- 11420879 TI - Severe hepatitis treated with an artificial liver support system. AB - We designed an artificial liver support system (ALSS) including plasma exchange, charcoal hemoperfusion, plasma bilirubin absorption, charcoal plasma perfusion, hemofiltration and hemodialysis. We chose different methods or their combinations according to the patient's conditions. We investigated the effect of ALSS in 154 patients with severe hepatitis, 72 of whom survived. All data were analyzed by SPSS. The effectiveness of ALSS treatment was compared at different stages (i.e. early, middle and end stages). After each ALSS treatment, the liver function of these patients greatly improved, serum endotoxin and HBV-DNA concentrations were significantly decreased, and the serum concentration of aromatic amino acids (AAA) such as methionine decreased while BCAA/AAA ratio increased. Patients treated with ALSS in the early or middle stages of disease had much higher survival rates than patients in the end stage of disease. Thus, we concluded that ALSS is a reliable therapy for advanced liver diseases and treatment in early or middle stages is appropriate. PMID- 11420880 TI - Design and analysis of coupling methods for modular endoprosthetic systems as an alternative to the conical coupling. AB - Modular endoprostheses are often used in bone tumor management. However, the conical coupling that connects the various modules has several shortcomings. As an alternative, four new couplings have been developed. To find out if they have sufficient strength and show no movement during loading, each coupling was analysed using the finite element method. Bolt force and friction coefficient was varied to examine their influence. From the analysis it was concluded that coupling B, a dovetail coupling, meets all requirements and is the best alternative to the conical coupling. Sensitivity to bolt force and friction coefficient is very limited. PMID- 11420881 TI - Requirements for an artificial intervertebral disc. AB - Intervertebral disc degeneration is an important social and economic problem. Presently available artificial intervertebral discs (AIDs) are insufficient and the main surgical intervention is still spinal fusion. The objective of the present study is to present a list of requirements for the development of an AID which could replace the human lumbar intervertebral disc and restore its function. The list addresses geometry, stiffness, range of motion, strength, facet joint function, center of rotation, fixation, failsafety and implantation technique. Date are obtained from the literature, quantified where possible and checked for consistency. Existing AIDs are evaluated according to the presented list of requirements. Endplate size is a weak point in existing AIDs. These should be large and fit vertebral bodies to prevent migration. Disc height and wedge angle should be restored, unless this would overstretch ligaments. Finally, stiffness and range of motion in all directions should equal those of the healthy disc, except for the axial rotation to relieve the facet joints. PMID- 11420882 TI - Preparation and characterization of SNO-PEG-hemoglobin as a candidate for oxygen transporting material. AB - Acellular hemoglobin (Hb) derivates developed as oxygen carriers are known to cause hypertensive reactions due to their nitric oxide (NO) scavenging action. To modulate this undesired activity, we have developed a new Hb derivative, s nitrosylated polyethylene glycol (PEG)-modified hemoglobin (SNO-PEG-Hb), which can deliver oxygen and NO. After human Hb was modified with PEG to increase its molecular weight, the free sulfhydryl groups of Hb were s-nitrosylated with s nitrosoglutathione. Administration of unmodified Hb into anesthetized rats caused a hypertensive reaction, while s-nitrosylated Hb derivatives such as SNO-Hb and SNO-PEG-Hb did not raise blood pressure. The plasma half-lives of heme and NO bound to SNO-PEG-Hb were 11.5 and 2.4 hours respectively, indicating that the s nitrosylated Hb derivative may act as a slow-releasing agent for NO. Based on these findings, SNO-PEG-Hb is a useful candidate for a blood substitute and tool for oxygen therapeutics. PMID- 11420883 TI - The in vitro biosynthesis and stability measurement with agyl-glycuronide isoformes of the main metabolite of ipriflavone. AB - The formation and stability of 1-beta-glucuronide conjugate of the main metabolite of ipriflavone [7-(1-carboxy-ethoxy)-isoflavone] (CI)--were studied by using liver microsomes, hepatocytes, and isolated perfused liver of untreated and 3-methylcholanthrene (MC) treated dog and rat, and human liver microsomes. MC treatment enhanced the rate of conjugation twice as much as that of the control in the microsomes of both dogs and rats. Conjugation of CI by microsomes results in two metabolites, both sensitive to pH and temperature. Other two glucuronide forms appeared in experiments with hepatocytes and perfused liver. Mass spectrometry supported. The conclusion, assumption that both metabolites produced by microsomes are glucuronide conjugate isoforms of CI, and that they could be distinguished according to the intensity of peaks on FAB-MIKE spectra. The beta glucuronidase enzyme hydrolysed only the 1-beta-glucuronide isomer, the other, migrated form remained unchanged. D-saccharic-acid-1,4-lactone, a specific inhibitor of beta-glucuronidase enzyme, decreased the rate of enzymatic cleavage. Standard curves of CI were prepared by HPLC, and 1-beta-CI-glucuronide was quantified according to the amount of CI formed by hydrolysis. The stability of conjugates greatly depends on pH and temperature, and the rate of degradation and isomerization is sensitive to the value of both. Lowering the pH from 7.4 to 5.0 and the temperature from 37 degrees C to 18 degrees C increased the stability of glucuronides. Increasing the pH to 12.0 results in very rapid acyl migration and hydrolysis. PMID- 11420884 TI - Uridine 5'-diphosphoglucuronic acid (UDPGLcUA) in the human fetal liver, kidney and placenta. AB - The endogenous concentration of uridine 5'-diphosphoglucoronic acid (UDPGLcUA), the endogenus substrate of UDP-glucuronosyltransferase, was measured in the human fetal and adult liver and kidney and in the placenta. The concentrations (mumol/Kg wet weight) of UDPGLcUA were 59.4 +/- 11.3 (fetal liver), 301 +/- 119 (adult liver), 11.9 +/- 3.2 (fetal kidney), 17.4 +/- 3.0 (adult kidney), 17.8 +/- 1.8 (mid-term placenta) and 17.0 +/- 1.7 (term placenta). UDPGLcUA is present in the human fetal liver at a concentration 5-fold lower than in the adult liver indicating a potential limiting factor for glucuronidation ind the human fetus. PMID- 11420885 TI - Mefloquine pharmacokinetics in healthy subjects and in peptic ulcer patients after cimetidine administration. AB - The pharmacokinetics of orally administered mefloquine were determined in six healthy male subjects and in six ulcer patients before and after a 3-day course of cimetidine (400 mg morning and evening). Peak plasma concentrations Cmax and AUC0-infinity were similarly and significantly (P < 0.05) increased after cimetidine pretreatement in both healthy subjects and peptic ulcer patients Cmax was increased by 42.4% and 20.5% while AUC0-infinity was increased by 37.5% in healthy and peptic ulcer subjects respectively. The values of t1/2ab absorption and t1/2 beta elimination, total crearance CLT/F and volume of distribution were altered to varying levels after cimetidine treatment but the changes were not statistically significant in both healthy and peptic ulcer subjects. The established long t1/2 beta and this apparent interaction between mefloquine and cimetidine which resulted in increased mefloquine plasma concentration might be of clinical significant in patients with neurological/psychiatric history. PMID- 11420886 TI - Absence of significant pharmacokinetic and pharmacodynamic interactions between artemether and quinoline antimalarials. AB - The study was carried out to investigate the pharmacokinetic and pharmacodynamic interactions between artemether (ARTEM) and quinoline antimalarials namely mefloquine (MQ), quinine (QN) and primaquine (PQ) when given concurrently. A randomised comparative, seven way cross-over design was performed in eight healthy male Thais following the administrations of seven drug regimens on seven occasions i.e. a single oral dose of ARTEM (300 mg), or MQ (750 mg), or QN (600 mg), or PQ (45 mg) alone, or the combination of ARTEM (300 mg) with MQ (750 mg), or QN (600 mg), or PQ (45 mg). All clinical and laboratory parameters were normal in all subjects, before, during and after the study. The eight subject experienced no adverse effect after ARTEM, QN, PQ alone regimens, or combination of ARTEM with QN and PQ. After administration of MQ in either occasion, 3 subjects had weakness, nausea, abdominal pain, and diarrhoea; one subject complained of dizziness. All symptoms were mild and occurred during the first day of MQ administration. The fitting of the concentration-time curves of ARTEM, QN and PQ, to a one-compartment model with first order absorption yielded satisfactory results in all subjects. The best fit model for MQ was two compartment model with first order absorption. The pharmacokinetics of all investigated drug, when given alone or in combination were not significantly different. PMID- 11420887 TI - Prevention by theophylline of beta-2-receptor down regulation in healthy subjects. AB - Adrenergic down-regulation can occur rapidly in many tissues. Therefore beta 2 agonists might have a rapidly decreasing effect in time, which is a potential problem for the treatment of bronchial asthma. This in vivo study tested the hypothesis that theophyline can prevent adrenergic down-regulation. A randomised, double blind, placebo-controlled cross-over study was performed in eight healthy subjects. Terbutaline concentration-effect relationships were studied before and after one week of dosing of terbutaline, with or without theophylline. Slow release terbutaline 5 mg daily was administered for 7 days in combination with either placebo or slow-release theophylline. Concentration-effect relationships of terbutaline after a single subcutaneous injection were studied before and after the 7 day terbutaline treatment. Eosinopenia and hypokalemia were the systemic effect parameters. Terbutaline concentration-time courses were described with a two-compartment model and those of theophylline with a polynomial equation. A hypothetical effect compartment model was applied to link terbutaline plasma concentration via an Emax model to the studied effects. The interaction of theophylline and terbutaline was described with a non-competitive pharmacodynamic model. After one week of oral terbutaline, the mean EC50 (ng/L) of terbutaline increased for the eosinopenia from 1.87 +/- 1.66 to 3.78 +/- 2.18 (+102%) (p = 0.012) with placebo, and to 2.73 +/- 1.99 (+46%) (p = 0.025) with theophylline; for the hypokalemia the EC50 increased from 4.70 +/- 2.91 to 8.52 +/- 7.26 (+81%) (p = 0.012) with placebo, and to 5.64 + 2.59 (+20%) (p = 0.16) with theophylline. The results indicate that the non-specific phosphodiesterase inhibitor theophylline can prevent terbutaline-induced adrenergic down-regulation to a substantial degree. PMID- 11420888 TI - Effect of vigabatrin and gabapentin on phynytoin pharmacokinetics in the dog. AB - The study was aimed at investigating whether or not the kinetics of intravenously administered phenytoin (PT) was altered by oral administration of vigabatrin (VGB) or gabapentin (GBP). A group of five beagle dogs were given a daily dose of PT (12 mg/kg, i.v.) for a period of 1 week. On day 8, plasma samples were serially collected over 24 hr. after administration of the PT dose. PT administration was continued with oral supplementary dose of VGB (60 mg/kg) for another week and then plasma samples were collected for analysis of PT levels. The same protocol was followed for the PT (12 mg/kg, i.v.)-GBP (300 mg caps., p.o.) study on a separate group (n = 5) of dogs. Orally administered GBP did not significantly alter the pharmacokinetic parameters of parental PT. VGB, however markedly changed the drug's kinetics as evidenced by a 31% (P = 0.015) reduction in total body clearance (CL) and increase of over 45% in half-life (t1/2), (P = 0.013) and area under the plasma PT concentration-time curve (AUC), (P = 0.044). GBP does not appear to have any pharmacokinetic interaction with PT, while coadministration of VGB and PT results in marked reduction in systemic clearance of the latter in the dog. PMID- 11420889 TI - Pharmacokinetics of reboxetine in healthy, elderly volunteers. AB - The pharmacokinetic characteristics of reboxetine, a unique selective noradrenaline reuptake inhibitor (selective NRI) for the treatment of depression, were studied in 12 healthy, elderly volunteers (mean age 81 years +/- 9 years). All subjects received a single 4-mg dose of reboxetine, and plasma reboxetine concentrations were measured by HPLC. Reboxetine was well tolerated by all subjects. Exposure to reboxetine was higher in this group of very elderly subjects, compared with data obtained in a similar study of young, healthy volunteers. Cmax in the elderly was 271 +/- 86 ng/ml, compared with 111 +/- 28 ng/ml in the young subjects after a single 4-mg dose, although in both groups Cmax was observed after 2 h. The AUC infinity was nearly four times that in the younger subjects (8345 +/- 3107 ng.h/ml vs. 2106 +/- 881 ng.h/ml) and the t1/2 was twice as long (24 +/- 6 h vs. 12 +/- 3 h). Renal clearance was also reduced. Reboxetine 8-10 mg/day has been effective and well tolerated in clinical trials in non-elderly depressed patients. The increased exposure to reboxetine observed in our very elderly subjects supports a reduction of the starting dose to 4 mg/day (in two divided doses) in the elderly. PMID- 11420890 TI - Disposition of amiodarone in rats after single and multiple intra-peritoneal doses. AB - The pharmacokinetics of amiodarone was studied after single and multiple dosing in two groups of male Wistar and Albino rats. The first group (40 rats) received a single intraperitoneal (i.p.) dose of amiodarone (100 mg/kg) and 4 rats sacrificed 1, 2, 4, 6, 12, 18, 24, 36, 48 and 72 hours post dosing. The second group (42 rats) received amiodarone (50 mg/kg, i.p., daily) for 5 days a week for 5 weeks and 6 rats were sacrificed at 1, 2, 3, 4, 5, 6 and 8 weeks. Rats of both group were sampled for blood, heart, lung and fat and the concentrations of amiodarone in these samples were determined using. The elimination of amiodarone from plasma after single dose followed a biphasic pattern with a terminal half life of 54.7 +/- 8.2 hours. The concentrations of amiodarone in the tissues were halved within 26.8, 34.9 and 37.45 hours in the heart, lung and fat, respectively. The average concentrations of amiodarone in plasma, heart, lung and fat after single dose were 1.24 micrograms/ml, 1.73 micrograms/mg, 7.61 micrograms/mg and 29.01 micrograms/mg, respectively. The concentration of amiodarone after multiple dosing were halved within 8.4, 5.5, 6.4 and 9.8 days, for the plasma, heart, lung and fat, respectively. The average concentrations of amiodarone in plasma, heart, lung and fat during multiple doses were 0.97 microgram/mg, 7.63 micrograms/mg and 65.01 micrograms/mg respectively. In conclusion, after multiple dosing, the elimination half-life of amiodarone and its fat contents were 3.7 and 2.8 times greater than that after single dosing. The excessive amount of amiodarone observed in fat tissues after multiple dosing is probably the reason for the prolonged elimination half-life. Based on the elimination half-lives data, the time to steady state is about two weeks and the drug should be withheld for less than a mont if a patient required discontinuation because of serious adverse effects. PMID- 11420891 TI - Bioinversion of ibuprofen enantiomers after administration in dogs: estimation of a novel index. AB - This study compares the pharmacokinetics and bioinversion of two chemical forms of ibuprofen administered intravenously or orally. Dogs were given the free acid form of the S(+) isomer p.o. or i.v., or the racemate, as the free acid or sodium salt, p.o., in a cross-over design. The main kinetic parameters were calculated and formation and bioinversion curves plotted. The values of Cmax, Tmax and AUC were higher for the S(+) isomer. The percentage bioinversion averaged between 35 70% according to the form. This study proposes a new index for the calculation of bioinversion, independently of any i.v. administration, and confirms its self limiting nature. PMID- 11420892 TI - Co-administration of melatonin reverses the tin-protoporphyrin (SnPP) induced decline of cytochrome P450 content in vivo in rats. AB - Melatonin (N-acetyl-5 methoxytryptamine) is a low molecular weight antioxidant and is an endogeneous defense system against the deleterious actions of the extremely reactive hydroxyl radical. Among the enzymes that participate in the antioxidant functions is cytochrome P-450, a stalwart of the detoxification system in the body. Our results revealed that tin-protoporphyrin administration brought about a marked decline in cytochrome P-450 levels. This decline was, however, reversed by the coadministration of the antioxidant, melatonin. Thus, the enhanced antioxidant status in melatonin-treated rats may act as a protective mediator of various pharmacological functions altered during tin-protoporphyrin (an antihyperbilirubemenic agent) administration to Wistar rats. PMID- 11420893 TI - Pharmacokinetics of midazolam in critically ill pediatric patients. AB - Midazolam is frequently used to produce sedation in critically ill pediatric patients. We studied the pharmacokinetics of midazolam in 22 patients (age 8 days to 16 years). The intravenous infusion rate to produce sedation ranged from 49 385 mcg/kg/hr. The blood samples were obtained at steady-state and midazolam was measured by gas chromatography with electron capture. The steady-state plasma concentrations of midazolam ranged from 49-385 ng/mL. The total clearance, apparent volume of distribution, and elimination half-life ranged from 0.1-3.1 L/kg/hr, 0.2-3.5 L/kg, and 0.3-10.9 hours, respectively. The marked interpatient variability in pharmacokinetics explains in part, the substantial variation in dosage requirements of midazolam to produce sedation in critically ill pediatric patients. PMID- 11420894 TI - Pharmacokinetic interactions of Mentat with carbamazepine and phenytoin. AB - In the present study, Mentat, a herbomineral psychotropic preparation, was studied for its pharmacokinetic interaction with the commonly used anti-epileptic drugs, carbamazepine and phenytoin. The interaction of carbamazepine and phenytoin with Mentat was studied in rabbits. Thirty two rabbits were divided into four groups of eight each. Animals of Group I were treated with carbamazepine (50 mg/kg b.wt. p.o.), Group II were treated with carbamazepine (50 mg/kg b.wt. p.o.) + Mentat (500 mg/kg b.wt. p.o.), Group III were treated with phenytoin (50 mg/kg b.wt. p.o.) and Group IV were treated with phenytoin (50 mg/kg b.wt. p.o.) + Mentat (500 mg/kg b.wt. p.o.) for a period of 8 days. On day 0 and day 8, plasma carbamazepine and phenytoin levels were estimated at different time intervals. A simultaneous treatment with Mentat resulted in a significant increase in plasma AUC of carbamazepine as well as phenytoin as compared to carbamazepine or phenytoin alone. Cmax and Tmax of carbamazepine and phenytoin also were evaluated. The results suggest that co-administration of Mentat could improve the effectiveness of anti-epileptic drugs due to the increased bioavailability of the latter. However, this has to be done with critical medical supervision to avoid any toxic reactions and preferably with therapeutic drug monitoring (TDM) which could also help in dose optimization. PMID- 11420895 TI - Pharmacokinetics of ketoprofen in rabbit after a single topical application. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly used by topical application in management of joint pain and inflammation. Little is known, however, about their pharmacokinetics, especially in the synovial compartment versus the plasma compartment, following topical administration. Ketoprofen, a NSAID, was delivered by a single topical application (KETUM 2.5% gel) on the rabbit knee-joint region of one hind limb. Concentrations of ketoprofen were measured in plasma, synovial fluid, joint capsule and synovial fat tissue at 2, 4, 6 and 12 hours after application. Whatever the time period after application, ketoprofen concentrations in synovial fluid were much higher than in plasma. The time-course of the decrease in ketoprofen plasma concentrations was more rapid than that in synovial fluid. Similarly, concentrations in joint capsule were higher than those found in synovial fat tissue. Finally, while ketoprofen concentrations decreased rapidly in plasma and in synovial fat tissue, concentrations in joint capsule and particularly in synovial fluid were more sustained. The increase in residence time of ketoprofen in synovial fluid could be in favour of its efficiency in the management of joint pain and inflammation. PMID- 11420896 TI - Designing population pharmacokinetic studies: performance of mixed designs. AB - The interplay of the following factors: population design (PDN), the cost function in terms of maximum cost (Max. C) (i.e., maximum number of samples/sample size), sample size, and intersubject variability [restricted (30%) to moderate (60%)] on the estimation of pharmacokinetic parameters from population pharmacokinetic data sets obtained using mixed designs was investigated in a simulation study. A two compartment model with multiple bolus intravenous inputs was assumed, and the residual variability was set at 15%. The sample size (N) investigated ranged from 30 to 200 with the associated cost function varying accordingly with the five individual and sixteen population designs studied. Accurate and precise estimates of structural model parameters were obtained for N > or = 50 (Max. C > or = 150) irrespective of the intersubject variability (ITV) and PDN investigated. When ITV was 30%, all structural model parameters were well estimated irrespective of the PDN. Robust estimates of clearance and its variability were obtained for all N at all levels of ITV with Max. C > or = 90 (PDN > or = 4). Imprecise estimates of ITV in V1, V2, and Q were obtained at 60% ITV irrespective of N, PDN, or Max. C. Positive bias was associated with the estimation of variability in V1, V2, and Q with PDN < or = 4 (Max. C < or = 150). This was due in part to a greater proportion of subjects sampled only once. Correspondingly, residual variability was underestimated. It is of utmost importance to avoid this artifact by ensuring that at least a moderate subset of subjects contributing data to a population pharmacokinetic study contribute data more than once. Given a sample size and ITV, the cost function must be considered in designing a population pharmacokinetic study using mixed designs. PMID- 11420897 TI - The existence of sequence effect in cross-over bioequivalence trials. AB - A generic drug product (T) in order to be approved for marketing authorization a bioequivalence trial is required. In the trial the generic product is compared to the innovator product (R) in terms of the pharmacokinetic parameters AUC and Cmax. The regulatory requirement for bioequivalence is that the 90% confidence intervals for the ratio (T/R) of the generic to innovator product pharmacokinetic parameter averages lies within the limits (80%, 125%). The design of the trial is usually a two-period crossover. This design has the limitation that if the statistical analysis reveal significant sequence effect then the bioequivalence results may be biased and their interpretation is difficult. The sequence effect is confounding with the unequal residual effect and with the formulation by period interaction. Since the existence of the sequence effect questions the quality of the trial, the applicant should provide possible explanations and information on the subjects, the trial conditions, the clinical settings and the assay methodology. An additional statistical analysis on the data from the first period of the trial may support the bioequivalence. If it is proven that the sequence effect is a true effect then the generic may be approved for marketing authorization. PMID- 11420898 TI - Dynamic study about metal release from titanium miniplates in maxillofacial surgery. AB - It has been shown that titanium (Ti) has had dramatic success in many surgical procedures as a result of its excellent mechanical properties and resistance to corrosion. There is still concern, however, about the release of metal and controversy surrounding whether or not the plates should be removed after bone healing. This study has been conducted to investigate whether or not there is a relationship between duration of plating and metal release from Ti miniplates in maxillofacial surgery. A prospective cohort study design was used. The concentration of Ti, in the soft tissues covering the plates, was examined in all patients who underwent removal of Ti miniplates from January 1998 to April 1999 (51 cases). Inductively Coupled Plasma-Optical Emission Spectrometry was used to measure Ti. Total ti and soluble Ti levels were compared to duration of plating (ranged from 15 days to 3 years, mean = 8 months). Correlation coefficients and two-way ANOVA were data processed. The average amount of total Ti in the soft tissues surrounding the plates was 1306 micrograms/g dry tissue. The mean of soluble Ti was 0.53 microgram/g dry tissue. The results of this study do not support the existence of a relationship between duration of plating and total Ti (correlation coefficient = 0.093 (P > 0.1) nor soluble Ti (correlation coefficient = 0.009 (P > 0.1) in the soft tissue surrounding the plates. Moreover, the only independent factor of Ti release found was associated with mechanical constraints during surgery. Almost 100% of Ti is released during the osteosynthesis. Then Ti levels remain constant in the surrounding tissues. Most of the time, Ti seems to be clinically inert. Compared to the possible risks of a second operation, removal of Ti miniplates should not be a routine procedure except in the case of complaints from patients, particularly in the case of infection, hypersensitivity, dehiscence or screw loosening. PMID- 11420899 TI - Treatment of temporomandibular joint ankylosis with temporalis muscle and fascia flap. AB - This study sought to determine the efficacy of interpositional arthroplasty with temporalis muscle and fascia flap in the treatment of unilateral temporomandibular joint (TMJ) ankylosis in adults. This retrospective study of seven cases evaluated the postoperative results of interpositional arthroplasty on temporalis muscle and fascia flap in adults. The operative protocol for unilateral TMJ ankylosis entailed, (1) resection of ankylotic mass, (2) intraoral ipsilateral coronoidectomy, (3) contralateral coronoidectomy when necessary, (4) interpositional tissue transfer to the TMJ with temporalis muscle and fascia flap, (5) maxillomandibular fixation (MMF), and (6) early mobilization and aggressive physiotherapy. The results of this protocol were encouraging, while the functional results of interpositional arthroplasty on temporalis muscle and fascia flap were satisfactory. The findings of this study support the use of temporalis muscle and fascia flap in adult patients with unilateral TMJ ankylosis. Early postoperative initial exercise, physiotherapy, and strict follow up play an important role in preventing postoperative adhesions. PMID- 11420900 TI - Effect of pumping with injection of sodium hyaluronate and the other factors related to outcome in patients with non-reducing disk displacement of the temporomandibular joint. AB - We retrospectively examined the effect of pumping with injection of sodium hyaluronate into the temporomandibular joint (TMJ) and the other factors influencing outcome in patients with non-reducing disk displacement of the TMJ. Fifty-nine patients underwent pumping with injection of sodium hyaluronate into the TMJ. As control, 62 patients were observed without any treatment. Both groups were observed for 12 months. The relation between outcome and the following clinical characteristics was also studied: sex, age, range of motion for maximal mouth opening, TMJ pain, TMJ noise, tenderness of masticatory muscles, locking duration, intercuspal occlusions, angle of posterior slope of articular eminence and degenerative bony changes of the condyle. Logistic regression analysis revealed that pumping with injection of sodium hyaluronate was related to a good outcome. Clinical characteristics of presentation significantly related to a good outcome were a large maximal mouth opening, a short locking duration, and a steep posterior slope of articular eminence. We conclude that pumping with injection of sodium hyaluronate into the TMJ is an effective treatment method for non-reducing disk displacement of the TMJ and that some clinical characteristics also influence outcome. PMID- 11420901 TI - A comparison of aesthetic, functional and patient subjective outcomes following lip-split mandibulotomy and mandibular lingual releasing access procedures. AB - Good access to the site of oral cancer is essential to allow for adequate three dimensional resection of the tumour. Splitting the lower lip in conjunction with a mandibulotomy offers excellent access to all areas of the mouth and pharynx, but this inevitably produces a facial scar and there is morbidity associated with the healing of the mandibulotomy. An alternative approach is the mandibular lingual releasing technique, which provides good access to the oral cavity and avoids the morbidity associated with lip-split mandibulotomy. The aim of this study was to compare aesthetic, functional and patient subjective outcomes between the two access procedures. One hundred and fifty patients had oral access procedures between 1992-95 (ninety lip-split mandibulotomy and sixty mandibular lingual release). Thirty patients fulfilled selection criteria (primary surgery as treatment, tumour size < 5.1 cm, anterior oral cavity tumours, and reconstructed with a radial forearm free flap) and 10 patients from each group were able to attend a review appointment for objective clinical assessment of their speech, tongue mobility, lip competence and temperomandibular signs. Using items from the University of Washington quality of life questionnaire patient subjective outcomes were assessed. Using standardised photographs the clinician and lay persons assessed the overall post-operative facial appearance. The patients also assessed their own facial appearance using a similar scoring method. Resection margins were similar in both groups and it would seem that both methods provide adequate access to the anterior oral cavity. Clinical examination showed no differences in function between the two access procedures. Although there was a small number, the lip-split mandibulotomy group reported significantly better speech, swallowing and chewing. Previous concerns about a possible detrimental effect on appearance following lip-split, were not borne out in this study. PMID- 11420902 TI - Tumours and tumour-like lesions of the oral and perioral structures of Nigerian children. AB - One hundred and fifty-eight children less or equal to 15 years were isolated from a retrospective 20-year (1979-1998) survey of oral and perioral tumours and tumour-like lesions in Nigeria. These children represent 16.8% of the cases seen. Lesions of children less or equal to 15 years were benign non-odontogenic (n = 70, 44.3%), benign odontogenic (n = 41, 25.9%) and malignant (n = 47, 27.7%). Predominantly Children between 11 and 15 years (n = 89, 56.3%) were afflicted and the male to female ratio was 1.4 to 1.0. Sites recorded were the jawbones, cheeks, gingival, forearm, testis and the spleen. The ratio of malignant lesions in Nigerian and African children appears to be higher than in the Caucasian population (2.4:1). PMID- 11420903 TI - The relationship between length of stay and health-related quality of life in patients treated by primary surgery for oral and oropharyngeal cancer. AB - A patient's length of hospital admission is simple to measure, whilst their health related quality of life (HRQOL) is much more challenging. Similar clinical and demographic factors may exist between an increased length of stay (LOS) and a worse HRQOL outcome. The aim of this study was to assess whether LOS could serve as a proxy marker of HRQOL. One hundred and thirty consecutive patients with previously untreated oral and oropharyngeal squamous cell carcinoma were recruited. All had primary surgery. The University of Washington questionnaire (UW-QOL) and the European Organisation for Research and Treatment core cancer questionnaire (EORTC C30) were self-completed preoperatively, after 6 months and 1 year. The median LOS following laser/primary closure/split skin grafts (21 patients) was 2 days compared to 16 days following microvascular free flap reconstruction (105 patients). In the free flap group the best predictors of LOS were age, tumour size and early medical/surgery complications. Patients with a LOS greater than 16 days (median) reported significantly worse chewing (P = 0.008), swallowing (P = 0.002) and cumulative UW-QOL score (P = 0.01). No significant differences were seen in the EORTC C30. Length of stay is potentially a useful indicator of health related quality of life because it is linked by tumour size, however, the relationship is confounded by age, which tends to influence length of stay more than health related quality of life. PMID- 11420904 TI - Patient compliance to instructions after oral surgical procedures. AB - The purpose of this study was to evaluate patient compliance to postoperative instructions from the oral surgeon. A total of 180 patients who underwent minor oral surgical procedures anonymously answered a questionnaire 1 week postsurgery. This questionnaire was used to evaluate how patients remembered the mode of transmission of instructions (written, verbal), their compliance to the postoperative instructions and to the antibiotic treatment and their comments. Of the patients, 40% did not remember receiving both written and verbal instructions, 36% remembered only the written instructions. Twelve percent did not comply with the postoperative instructions regarding mouthwashes and 67% did not comply with the antibiotic prescriptions; 43% took more antibiotics and 31% less, and 4% who did not receive an antibiotic prescription took them on their own initiative. The academic level of the patients did not influence patient compliance. Verbal and written recommendations worded simply with details are important for patient compliance and reduced postoperative stress and complaints. Better instructions on antibiotics use are necessary to ameliorate patient compliance to the antibiotic treatment. PMID- 11420905 TI - Comparison of different distraction rates in the mandible: an experimental investigation. AB - This study evaluated bone distracted in rabbit mandibles at different intervals and with different daily rates of distraction histologically with the goal of attaining a universally accepted distraction protocol. Osteogenesis was investigated in mandibles distracted at different rates in 24 New Zealand rabbits using a custom-made submerged distractor. Distraction was started on the third postoperative day for a total of 10 mm. The animals were divided into four groups each containing six rabbits. Group 1 was distracted 0.5 mm twice a day; Group 2 was distracted 1.0 mm once a day; Group 3 was distracted 1.0 mm twice a day and Group 4 was distracted 2.0 mm once a day. All the animals were sacrificed 6 weeks after completion of distraction. Half of the distracted mandibles were decalcified for H&E staining and polarized light microscopy studies. Sections of the undecalcified half of the samples were stained with Goldner's stain. The results indicate that a distraction rate of 1.0 mm per day produced the best osteogenesis among the tested rates. There was no great difference in osteogenesis between 1.0 mm once a day and 0.5 mm twice a day. However, 0.5-mm distraction may result in immature bone healing. Distraction of 1.0 mm twice a day resulted in incomplete osteogenesis, while distraction of 2.0 mm once a day resulted in fibrous union. It is clear from these results that a shorter period of device fixation should be achieved by methods other than rapid distraction. PMID- 11420906 TI - Effect of calcium sulphate on the rate of osteogenesis in distracted bone. AB - The effect of applying resorbable calcium sulphate material to newly distracted bone to hasten osteogenesis and consolidation was investigated using 24 New Zealand rabbits. The animals were divided into four groups each containing six rabbits. Groups 1 and 2 were test groups and Groups 3 and 4 were controls. A custom-made submerged distractor was used to distract the rabbit mandibles. The distractor was first used on the third postoperative day at a rate of 1 mm/day for a total of 10 mm. After the completion of distraction, a medical grade resorbable calcium sulphate mixture was applied on the distracted bones in the test groups, whereas the distracted bones in the control groups were exposed postdistraction and closed without intervention. Groups 1 and 3 were sacrificed 11 days postdistraction and Groups 2 and 4 were sacrificed 21 days postdistraction. Undecalcified sections of the distracted bone were examined. Half of the distracted mandibles in each group were stained with Goldner's trichome stain to identify the amount and maturity of the new bone in the distraction zone. The other half of each group was stained with von Kossa stain to identify the amount of calcification and calcium deposition in the distracted bone. Osteogenesis was seen as early as 11 days postdistraction in test animals. Rich lamellar bone deposition was noted in Group 2 (21 days postdistraction +CaSO4). Calcification of the newly formed bone was predominant in the distraction zone in the test groups, especially Group 2, compared to the control groups, which showed incomplete bone deposition and calcification. In conclusion, the application of calcium sulphate to newly distracted bone increased the rate of osteogenesis and calcification. PMID- 11420907 TI - Mineralization dynamics of regenerate bone during mandibular osteodistraction. AB - Distraction osteogenesis offers an alternative approach for the correction of craniofacial deformities, yet little substantive data is available regarding mineralization of the forming regenerate. The purpose of this study was to evaluate, by plain film radiography and digital subtraction radiography, the mineralization dynamics of regenerate bone during osteodistraction. Thirteen skeletally mature male beagle dogs underwent 10 mm of bilateral mandibular distraction osteogenesis, at which time the bone was allowed to consolidate for either 4, 6, or 8 weeks. The tissues were then evaluated clinically and radiographically. The results demonstrate that several different types of regenerate are evident during mineralization of the distraction gap. A classification system is presented based on the length, width, and density of the mineralizing regenerate, as well as the presence or absence of an interzone. It appears that important factors for determining regenerate stability are the presence or absence of an interzone, as well as the width of the regenerate relative to the host bone segments. Finally, in cases where an interzone is present, subtraction radiography may provide a means of evaluating the level of mineralization within the interzone as well as the dynamics of mineralization over the course of distraction and consolidation. PMID- 11420908 TI - Renal morphology in connexin43 knockout mice. AB - Connexins (Cx) are a family of proteins that constitute the intercellular membrane channels of gap junctions. These junctions permit intercellular movement of ions and other molecules between cells, a property vital to organogenesis. Cx43 is a member of the family of channel-forming proteins that are essential for cell-cell communication of developmental signals. Studies demonstrate that Cx43 is observed in mesenchymal cells of 12-day gestation mouse kidney, a crucial period of renal development. In order to study the significance of Cx43 on renal developmental morphology, we evaluated the kidneys of embryos lacking the gene encoding for Cx43. Polymerase chain reaction (PCR) from tail specimens identified wild-type (WT), heterozygote (HT) and knockout (KO) progeny. In situ RT-PCR displayed abundant Cx43 staining in glomeruli, vasculature, and tubules in kidneys obtained from WT progeny. In contrast, Cx43 expression was completely absent in kidneys isolated from the KO. Renal histology in all three groups displayed no significant differences. Renal size was similar and there was no evidence of dysplasia or cyst formation in the KO. Our results indicate that absence of Cx43, heretofore considered essential for renal development, does not affect early renal morphological development. PMID- 11420909 TI - The effect of vitamin A on the course of renal ablation nephropathy. AB - Renal scarring due to pyelonephritis was shown to improve in rats given vitamin A. We evaluated the effect of vitamin A in a renal ablation nephropathy model. Four groups, each including 7 rats with 5/6 nephrectomy, were formed: group I (no vitamin A), group II (60 kIU vitamin A), group III (120 kIU vitamin A), and group IV (180 kIU vitamin A). Four sham-operated rats comprised the control group. After 6 weeks of 5/6 nephrectomy, the rats were sacrificed and serum creatinine, vitamin A, and beta-carotene levels were determined in addition to histopathological evaluation of the remnant kidneys. The tubulointerstitial and glomerular changes were graded as "0-3" and "0-5" respectively, in accordance with the severity of the lesions. Tubulointerstitial score (TIS), mean glomerulosclerosis score (MGS, arithmetical mean of the sclerosis scores of 100 glomeruli), and severity of glomerulosclerosis index (SGI, ratio of the number of glomeruli with grade > or = 3 sclerosis to the total number of glomeruli examined) were calculated for each rat. Serum creatinine levels were higher in the study groups than the control rats (P < 0.05), but there was no significant difference between the study groups (although the levels increased as the dose of vitamin A increased). Serum vitamin A levels were significantly higher in the groups given vitamin A than the control rats and group I (P < 0.05). In addition, serum vitamin A levels increased significantly in parallel to increasing doses of vitamin A (P < 0.05). Serum beta-carotene levels did not differ between the groups, except for group II, which had lower levels than controls (P = 0.01). MGS and SGI were significantly higher in the study groups than control rats (P < 0.05), but did not differ between the study groups. Study and control rats were not different with respect to TIS, but there was a difference between the control group and group III (P = 0.04). Group II had the lowest MGS, SGI, and TIS scores among the study groups. When all the rats were considered together, vitamin A levels did not correlate with the MGS and SGI, but correlated positively with the TIS (r = 0.391, P = 0.027). beta-Carotene levels also did not correlate with the MGS, SGI, and TIS. In conclusion, vitamin A administration did not significantly affect the clinical and pathological course of renal ablation nephropathy in rats. Furthermore, higher doses of vitamin A might even damage renal tissue. PMID- 11420910 TI - Postnatal disappearance of type A intercalated cells in carbonic anhydrase II deficient mice. AB - Despite chronic acidosis, collecting ducts in adult carbonic anhydrase II deficient (CAD mice) are depleted of intercalated cells, including those of type A, which are acid-secreting cells. We hypothesized that this depletion could occur during postnatal development. Principal cells were identified by immunofluorescence using an antibody to rat aquaporin-2 (AQP-2), and type A intercalated cells using an antibody specific for anion exchanger (AE1). In CAD mice the proportion of AQP2-positive cells, normal at 11 days, increased progressively in the cortical (CCD) and outer medullary collecting duct (OMCD), to reach almost 100% in the OMCD in adults. The percentage of AE1-positive cells in the OMCD of CAD mice decreased by half by 6 weeks of age and further by adulthood. In controls, however, the proportion of AQP2-positive cells and that of AE1-positive cells in the OMCD remained stable after 10 days of age. AE1 positive cells accounted for the majority of intercalated cells in the OMCD. The mechanisms leading to selective postnatal cell depletion in the collecting duct in CAD mice remain to be determined. PMID- 11420911 TI - Histopathological evidence of poor prognosis in patients with vesicoureteral reflux. AB - Patients with vesicoureteral reflux (VUR) often develop reflux nephropathy with focal segmental glomerular sclerosis (FSGS), although the exact mechanisms leading to the development of this complication are unknown. To determine the early changes in glomeruli of VUR patients that ultimately cause poor renal outcome, we examined morphometrically renal biopsies of 16 young patients (age 10 20 years) with VUR at baseline pre-operatively. Patients were divided into two groups, those who subsequently showed good prognosis and those with poor renal prognosis at the end of a 10-year follow-up period. Patients with poor prognosis had worse proteinuria and lower creatinine at baseline than those with good prognosis. We also examined 40 age-matched control cases with previous temporal microhematuria and/or proteinuria but normal renal function and histology. Although the mean diameter of glomerular capillary did not change in VUR cases irrespective of prognosis, glomerular capillary length increased by 125% in cases with good prognosis, and 335% in cases with poor prognosis (P < 0.01). Cystically expanded capillaries, with diameter > or = 95% of that in age-matched control, were detected in five of eight patients with poor prognosis, but only in one of eight patients with good prognosis. In VUR, the number of podocytes/capillary diminished with increased length of the capillaries. Tuft adhesion to Bowman's capsule and podocyte detachment were primarily found in patients with poor prognosis. Our results suggest that lengthening of glomerular capillaries in young patients with VUR is a compensatory reaction to hyperfiltration. The appearance of cystic capillary expansion, podocyte detachment and/or tuft adhesion to Bowman's capsule in such glomeruli may be important indicators of renal prognosis in patients with VUR. These changes may lead to FSGS due to podocyte injury in patients with VUR, with subsequent deterioration of renal function. PMID- 11420912 TI - Early urinary changes in Tamm-Horsfall protein and epidermal growth factor in diabetic children. AB - Both glomerular and tubular markers have been used to follow diabetic nephropathy. However, neither albumin nor proximal tubular markers have proven useful in prepubertal diabetes. Hence we studied two markers derived from the distal tubular cells, Tamm-Horsfall protein (THP) and epidermal growth factor (EGF). The urinary excretion of THP and EGF was examined in samples obtained during the first 20 days and 1 year after diagnosis of diabetes in children aged 4-15 years. Fourteen children without and 18 with ketonuria were examined, and 17 age-matched healthy children participated as controls. The excretion rate of EGF was increased at diagnosis, while that of THP was not. After 20 days of treatment the excretion of EGF had normalized, while the excretion of THP was decreased. Similar results were obtained after 1 year. In conclusion, in spite of good metabolic control a reduced excretion of THP persisted for at least 1 year after the diagnosis of diabetes. Whether the finding of reduced excretion of THP has any biological significance awaits further study. PMID- 11420913 TI - Angiotensin-II and endothelin-1 levels in children with renoprival hypertension. AB - Children with end-stage renal disease (ESRD) often remain hypertensive despite bilateral nephrectomy and aggressive fluid removal on hemodialysis. We speculated that an extrarenal source of renin might contribute to the release of "tissular" angiotensin-II (AT-II) generating hypertension in anephric patients. At the same time, experimental evidence supports that peripheral AT-II vasoconstrictive effect is likely mediated by endothelin-1 (ET-1). Thus, it is conceivable that hypertension in ESRD patients may be due, in part, to a cascade involving vascular production and secretion of AT-II and ET-1. In order to establish the relationship between AT-II, ET-1, and blood pressure we performed a pilot study to measure predialysis systolic and diastolic blood pressures (SBP and DBP, respectively) and serum AT-II and ET-1 levels in 12 anephric children receiving hemodialysis. Predialysis AT-II and ET-1 levels were similar in all patients, and neither value correlated with their mean SBP or DBP. In patients with postdialysis hypertension, there was no correlation between predialysis AT-II and ET-1 plasma levels. We therefore find no evidence to suggest that vascular mediated AT-II and/or ET-1 contributes significantly to hypertension in anephric patients. PMID- 11420914 TI - Does albumin and furosemide therapy affect plasma volume in nephrotic children? AB - Albumin infusions transiently increase plasma volume (PV) and oncotic pressure, and may restore diuretic responsiveness in nephrotic edema. To determine if albumin and furosemide therapy have an effect on PV in nephrotic children, 14 severely edematous children with minimal change nephrotic syndrome were evaluated with standard clinical parameters (heart rate, blood pressure, body weight, pretibial edema, abdominal circumference) and echocardiography [inferior vena cava index (IVCI), inferior vena cava collapsibility index (IVCCI)] before, 1 h and 24 h after albumin (20%, 0.5 g/kg, 1 h) and furosemide (2 mg/kg, i.v.) therapy. An increase in IVCI (P < 0.05), decrease in IVCCI (P < 0.05), edema (P < 0.005), and hematocrit (P < 0.005) were statistically significant 1 h after albumin and furosemide therapy, with a transient effect 24 h later. Body weight (P < 0.005), abdominal circumference (P < 0.05), and edema (P < 0.005) decreased significantly at 24 h. It is concluded that albumin and furosemide therapy increases PV transiently in nephrotic edema, returning to baseline values at 24 h with a decrease in body weight, abdominal circumference, and edema. PMID- 11420915 TI - Henoch-Schonlein purpura in Wiskott-Aldrich syndrome. AB - Wiskott-Aldrich syndrome (WAS) is a rare immune deficiency disease. Sialophorin glycosylation is defective in WAS. Although it is not very common, renal involvement including IgA nephropathy (IgAN) was reported. Abnormal glycosylation plays a key role in the pathogenesis of IgAN. We present an 8-year-old boy with WAS who had recurrent episodes of Henoch-Schonlein purpura with renal involvement following upper respiratory tract infections. His renal function did not deteriorate. Both IgAN and WAS have glycosylation defects, but there must be some other factors (genetic and environmental) to explain their rare association. PMID- 11420916 TI - Role of Tc-99m DMSA scintigraphy in the diagnosis of culture negative pyelonephritis. AB - The aim of this work was to evaluate prospectively the proportion of children with a clinical and biological presentation of acute pyelonephritis, abnormal Tc 99m dimercaptosuccinic acid (DMSA) scintigraphy and negative or equivocal urine cultures. All patients aged 6 weeks to 15 years suspected of acute pyelonephritis (APN) were admitted to the Paediatric Department and underwent Tc-99m DMSA scintigraphy within 3 days after admission and at 6 months. Of 166 patients enrolled in the study, 15 (9%) had negative or equivocal urine culture despite clinical and scintigraphic evidence of APN. Of these 15 children, renal ultrasound was normal in 7 patients and vesicoureteric reflux was found in 9 patients. Control DMSA 6 months after acute episode showed the disappearance of cortical lesions in eight and partial improvement in four patients. In this prospective series, as many as 9% of patients with APN would have been missed on the basis of equivocal or negative urine cultures. It is suggested that Tc-99m DMSA scintigraphy should be performed in children with severe infection without clear aetiology, especially in those with abnormal urinalysis. PMID- 11420917 TI - Nephronophthisis and ulcerative colitis in siblings: a new association. AB - Nephronophthisis (NPH) is a chronic tubulointerstitial nephritis leading to terminal renal insufficiency. The disease is heterogeneous, but usually the inheritance pattern is autosomal recessive. In 80% of cases, the disease is caused by a homozygous deletion in NPHP1 gene in chromosome 2q13. Ulcerative colitis is an inflammatory bowel disease with chronic diarrhea, rectal bleeding and characteristic histological findings. Its etiology is suggested to be multifactorial, consisting of genetic susceptibility and unknown exogenous factors. We present two siblings with NPH and ulcerative colitis. As NPH in this family is not linked to 2q13, this association may represent a new, syndromic form of NPH. PMID- 11420918 TI - Reversible nephrotoxicity after overdose of colloidal bismuth subcitrate. AB - Although toxicity due to acute and chronic use of bismuth salts is well known, nephrotoxicity after ingestion of colloidal bismuth has been reported in few cases so far. Here we report the first case of acute renal failure (ARF) due to colloidal bismuth subcitrate overdosage in childhood. A 2-year-old boy was admitted to the hospital 6 h after ingestion of 28 De-Nol tablets (colloidal bismuth subcitrate 8.4 g). On admission, physical examination was unremarkable and he showed no signs of encephalopathy. Initially gastric lavage was performed then appropriate fluid therapy was started. ARF associated with uremia and oliguria developed on day 2 and peritoneal dialysis therapy was prescribed on day 4 for 10 days. Blood and urine bismuth levels were 739 micrograms/l and 693 micrograms/l, respectively, 10 days after the pills had been taken. His urine volume gradually increased and plasma BUN and creatinine levels decreased during peritoneal dialysis. On day 20 post-admission, plasma BUN and creatinine were 14 mg/dl and 0.7 mg/dl, respectively. Blood bismuth levels were 96 micrograms/l on day 60 and 12 micrograms/l on day 105. Now the patient is well and has no problem. This case suggests that ARF may develop in children following colloidal bismuth subcitrate overdosage; the prognosis is good, and peritoneal dialysis may be useful in these cases. PMID- 11420919 TI - Steroid responsive nephrotic syndrome associated with bilateral renal dysplasia. AB - Renal dysplasia is characterized by hypoplastic kidneys that contain elements of primitive tubules. Patients may develop end-stage renal failure early in life. Nephrotic syndrome is one of the most common renal diseases in childhood and may occur in association with renal dysplasia. We report a case of a child with bilateral dysplastic kidneys and steroid responsive nephrotic syndrome (SRNS). An association between renal dysplasia with chronic renal failure and SRNS has not previously been reported in the English literature. PMID- 11420920 TI - Systolic hypertension in children: benign or beware? AB - Systolic blood pressure (SBP) has become the major criterion for the diagnosis, staging, and treatment of hypertension in adults, based on the epidemiology and pathophysiology of adult hypertension, linkage between SBP levels and disease, and benefits of treatment of isolated SBP hypertension. Although children do not typically suffer overt hypertensive disease, an accumulation of data suggests that SBP elevation is as important a factor in the morbidity of hypertension in children as in adults. SBP hypertension is more common in children, whether examining an unselected sample of patients by routine screening or a selected sample of referred hypertensive patients. Mild-to-moderate BP elevation in children is associated with increased left ventricular mass (LVM), with SBP more closely linked to LV morphology than diastolic blood pressure (DBP). Furthermore, SBP is associated with increased LVM even in patients with SBP within the "normal" range. Among hypertensive children, the reported prevalence of LVH ranges from 30% to 70%, and LV hypertrophy is more closely related to SBP than to DBP. These data suggest that treatment of hypertension should be directed at normalization of SBP, even when DBP is within the normal range. In addition, trials of anti-hypertensive medications in children should incorporate SBP hypertension into study inclusion criteria. PMID- 11420921 TI - Management of the initial episode of steroid-responsive nephrotic syndrome. PMID- 11420922 TI - Treatment of steroid-sensitive nephrotic syndrome. PMID- 11420923 TI - Pediatric renovascular hypertension. PMID- 11420924 TI - How can one apply rescue indomethacin therapy to a 1-month-old baby with antenatal Bartter syndrome in the case of severe vomiting? PMID- 11420925 TI - Issues and trends in instruction and deafness: American Annals of the Deaf 1996 to 2000. PMID- 11420926 TI - Literacy publications: American Annals of the Deaf 1996 to 2000. PMID- 11420927 TI - Evaluation of the dermatophyte test medium RapidVet-D. AB - The performance of the dermatophyte test medium (DTM) RapidVet-D was assessed using hair samples collected from experimentally infected guinea pigs. Three dermatophyte species were included in the study: Microsporum canis, Trichophyton mentagrophytes and Trichophyton equinum. DTM substrates were inoculated with infected hairs and scales, incubated at 18, 21, 24, 27 or 37 degrees C and examined daily for 15 days. The rapidity of colour change was clearly related to the incubation temperature and to the number of infected hairs deposited on the reactive substrates. With the optimum incubation temperature 27 degrees C, a systematic colour change could be observed only a few days post-inoculation: 3 days with M. canis infected hairs, 4 days with T. equinum and 5 days with T. mentagrophytes. PMID- 11420928 TI - TNF-alpha dependent NF-kappa B activation in cultured canine keratinocytes is partly mediated by reactive oxygen species. AB - The cytokine TNF-alpha plays a major role in inflammatory and immunological reactions of canine skin. With respect to a possible therapeutic modulation, we investigated the role of the transcription factor NF-kappa B and the involvement of reactive oxygen species (ROS) in the TNF-alpha signalling pathway in cultured canine keratinocytes. TNF-alpha treatment resulted in activation of NF-kappa B which was partially inhibited by the antioxidant alpha-lipoic acid. Using the cytochrome c reduction test no superoxide production could be detected in the supernatant of TNF-alpha stimulated keratinocytes. However, TNF-alpha dependent intracellular hydrogen peroxide production was demonstrated spectroscopically. With electron energy loss spectroscopy (EELS) significant hydrogen peroxide formation was detected in the mitochondria, the endoplasmic reticulum, the cytosol and partially on the plasma membrane of the keratinocytes. Hence, ROS possibly play an important role in the TNF-alpha signalling pathway leading to NF kappa B activation in canine skin. An adjunctive therapy with natural potent antioxidants modulating NF-kappa B overactivation in canine cutaneous inflammation may be of therapeutic benefit. PMID- 11420929 TI - MIB-1 immunoreactivity correlates with biologic behaviour in canine cutaneous melanoma. AB - The growth fraction of 68 canine cutaneous melanomas was determined by immunostaining with MIB-1, a monoclonal antibody to a Ki-67 epitope that recognizes all proliferating cells. Fifty tumours were classified histologically as benign and 18 as malignant. The Ki-67 proliferative index (percentage of positive cells over 500 neoplastic cells) was low (< 15%) in 55 cases and high (> or = 15%) in 13 cases. High Ki-67 proliferative index and histological malignancy were both associated with significantly poorer 2-year survival (P < 0.0001). However, the predictive value of the Ki-67 proliferative index (97%) was higher than the predictive value of classical histology (91%). The evaluation of the growth fraction by the Ki-67 proliferative index is highly predictive of the biological behaviour of canine cutaneous melanoma. PMID- 11420930 TI - Intradermal skin test reactivity to histamine and substance P is blunted in dogs with atopic dermatitis. AB - Skin reactivity to intradermal injections (0.1, 0.5 and 1 nM) of substance P (SP) was evaluated in 20 clinically normal dogs and 20 dogs with atopic dermatitis (AD). Saline and histamine were used as negative and positive controls, respectively. Wheal diameters were measured. Reactions were evaluated for erythema and induration and a subjective score, on a scale from 0 to 4+, was given. Evaluations were performed at 3, 5, 10, 15 and 30 min after the injections. Wheal diameters for histamine and SP injections were significantly smaller in dogs with AD compared with clinically normal dogs. In both groups, reactions to the various concentrations of SP were not significantly different from each other and were always smaller than histamine reactions. Erythema was not seen with SP injections. In addition, subjective scores for SP injections were significantly lower in dogs with AD compared with controls. The results of this study are similar to those reported in human medicine, where a role for SP in AD is proposed and desensitization of receptors to both SP and histamine is hypothesized. Further studies are needed to investigate the role of SP in the pathogenesis of canine AD. PMID- 11420931 TI - Induction of feline flea allergy dermatitis and the incidence and histopathological characteristics of concurrent indolent lip ulcers. AB - The objectives of this study were to characterize the role of intermittent vs. continual flea exposure in the development of flea allergy dermatitis (FAD) in cats, assess the accuracy of intradermal skin testing (IDST) and in vitro testing, and document the incidence and histopathological features of indolent lip ulcers. Ten flea-naive cats were divided into two groups. One group received intermittent flea exposure for 120 days. Thereafter, both groups of cats received continuous flea exposure for 120 days. In vitro testing for flea salivary antibody and IDST utilizing both whole flea antigen and flea salivary antigen were performed. Eight of 10 cats developed clinical signs of FAD within 3 months and five of these eight cats developed lip ulcers which where characterized histopathologically by ulceration with predominantly neutrophilic inflammation and surface bacterial colonization. There was no association between the presence or absence of clinical signs and positive IDST or in vitro results, and no difference in the development of clinical signs was noted between the two groups of cats. PMID- 11420932 TI - Evaluation of the clinical efficacy of marbofloxacin (Zeniquin) tablets for the treatment of canine pyoderma: an open clinical trial. AB - The efficacy and field safety of marbofloxacin (Zeniquin) for the treatment of superficial and deep bacterial pyoderma were evaluated. Seventy-two dogs were treated with 2.75 mg kg-1 of marbofloxacin orally once daily for 21 or 28 days. Sixty-two dogs (86%) had superficial pyoderma and 10 (14%) had deep pyoderma. A history of prior pyoderma was reported in 39/72 dogs. Pretreatment aerobic bacteriologic cultures of skin lesions were performed in 47 cases and the predominant pathogen isolated was Staphylococcus intermedius. Treatment was successful in 62/72 (86.1%) dogs, improvement was noted in 6/72 (8.3%) dogs and treatment failed in 4/72 (5.6%) dogs. Adverse effects associated with treatment included listlessness, anorexia, vomiting, soft stool, flatulence and polydipsia; these adverse effects were seen in only 6/81 dogs. Marbofloxacin was safe and effective for the treatment of superficial and deep pyoderma in dogs at the dosage used in this study. PMID- 11420933 TI - Antimicrobial activity of enrofloxacin against Staphylococcus intermedius strains isolated from canine pyodermas. AB - This study examined and compared the minimal inhibition concentrations (MICs) of enrofloxacin against 393 Staphylococcus intermedius strains isolated in France from canine pyodermas during three different years, 1995 (174 isolates), 1997 (101 isolates) and 1999 (118 isolates). The MICs of enrofloxacin against these strains ranged from 0.063 to 64 mg L-1, with MIC50 and MIC90 equal to 0.125 and 0.25 mg L-1, respectively. Two resistant strains were found, but only among isolates collected in 1999. The data show that resistance to enrofloxacin among S. intermedius strains is still rare in dogs, but the selection in vitro of variants in which the MICs were increased 4-16-fold after 10 serial passages in subinhibitory concentrations of enrofloxacin suggests that inappropriate use might favour the development of resistant strains in vivo. PMID- 11420934 TI - Feline atopy in three littermates. AB - This case report describes the history, clinical signs and diagnosis of a pruritic skin disease in three sibling cats living the same household. Clinical signs consistent with pruritus (i.e. hair pulling, hair loss, excessive grooming and face rubbing) were first noted when the cats were 6 months of age. The cats were treated for a possible ear mite and/or flea infestation; there was no response to treatment and clinical signs progressed. Although the presence of pruritus in a multiple cat household suggested an infectious or contagious aetiology, none could be identified. There was no improvement in clinical signs after a 60-day flea control trial, three treatments of ivermectin, an 8-week restricted diet or removal from the home for 10 days. A diagnosis of feline atopy was made on the basis of elimination of other causes of pruritus, consistent history and clinical signs, a positive intradermal skin test and response to therapy. PMID- 11420935 TI - Real time imaging of the production and movement of nitric oxide in the retina. PMID- 11420936 TI - Integration and segregation of visual signals by bipolar cells in the tiger salamander retina. PMID- 11420937 TI - Transmission at the photoreceptor synapse. PMID- 11420938 TI - Organization of ON- and OFF-pathways in the zebrafish retina: neurotransmitter localization, electrophysiological responses of bipolar cells, and patterns of axon terminal stratification. PMID- 11420939 TI - Retinal information processing and ambient illumination. PMID- 11420940 TI - Plasticity of AII amacrine cell circuitry in the mammalian retina. PMID- 11420941 TI - Neuromodulation of voltage-dependent K+ channels in bipolar cells: immunocytochemical and electrophysiological studies. PMID- 11420942 TI - Synaptic mechanisms shaping the light-response in retinal ganglion cells. PMID- 11420943 TI - Parallel processing in the mammalian retina: lateral and vertical interactions across stacked representations. PMID- 11420944 TI - Pre- and postsynaptic mechanisms of spontaneous, excitatory postsynaptic currents in the salamander retina. PMID- 11420945 TI - Physiological responses associated with kainate receptor immunoreactivity in dissociated zebrafish retinal neurons: a voltage probe study. PMID- 11420946 TI - GABA transporter function in the horizontal cells of the skate. PMID- 11420947 TI - Comparative anatomy of major retinal pathways in the eyes of nocturnal and diurnal mammals. PMID- 11420948 TI - Physiological and pharmacological characterization of glutamate and GABA receptors on carp retinal neurons. PMID- 11420949 TI - The GABAC receptors of retinal neurons. PMID- 11420950 TI - Cellular organization of the vertebrate retina. PMID- 11420951 TI - Retinoic acid, a neuromodulator in the retina. PMID- 11420952 TI - Properties of turtle retinal ganglion cell GABA receptors. PMID- 11420953 TI - The rhodopsin cycle: a twist in the tale. PMID- 11420954 TI - Insights into the rod rhodopsin regeneration process using the excised mouse eye. PMID- 11420955 TI - The response gradient along the rod outer segment: cGMP, age and calcium. PMID- 11420956 TI - The flash response of rods in vivo. PMID- 11420957 TI - Dark adaptation. PMID- 11420958 TI - Evaluation of the contributions of recoverin and GCAPs to rod photoreceptor light adaptation and recovery to the dark state. PMID- 11420959 TI - Light adaptation and contrast in the outer retina. PMID- 11420960 TI - Synaptic mechanisms of network adaptation in horizontal cells. PMID- 11420961 TI - Triphasic adaptation of teleost horizontal cells. PMID- 11420962 TI - Potassium conductances and the glutamate transporter in Muller cells of the turtle retina and their role in potassium siphoning. PMID- 11420963 TI - Some aspects of the oscillatory response of the retina. PMID- 11420964 TI - Light and circadian regulation of retinomotor movement. PMID- 11420965 TI - Circadian and efferent modulation of visual sensitivity. PMID- 11420966 TI - Circadian clock regulation of neuronal light responses in the vertebrate retina. PMID- 11420967 TI - Spinules and nematosomes in retinal horizontal cells: a "thorny" issue. PMID- 11420968 TI - Synaptic organization in the fly's optic lamina: few cells, many synapses and divergent microcircuits. PMID- 11420969 TI - Understanding retinal cell fate determination through genetic manipulations. PMID- 11420970 TI - Genetic and epigenetic analysis of visual system functions of zebrafish. PMID- 11420971 TI - Genetic analysis of initial and ongoing retinogenesis in the zebrafish: comparing the central neuroepithelium and marginal zone. PMID- 11420972 TI - Retinoic acid synthesis and breakdown in the developing mouse retina. PMID- 11420973 TI - Postnatal development of the rat retina and some of its neurotransmitter systems in vitro. PMID- 11420974 TI - The function of the cholinergic system in the developing mammalian retina. PMID- 11420975 TI - Legacy of the RCS rat: impact of a seminal study on retinal cell biology and retinal degenerative diseases. PMID- 11420976 TI - Retinal disease in vertebrates. PMID- 11420977 TI - Photoreceptor rescue in an organotypic model of retinal degeneration. PMID- 11420978 TI - Rod-cone interdependence: implications for therapy of photoreceptor cell diseases. PMID- 11420979 TI - Genes and diseases in man and models. PMID- 11420980 TI - Experimental retinal detachment: a paradigm for understanding the effects of induced photoreceptor degeneration. PMID- 11420981 TI - The origin of photo-oxidative stress in the aging eye. PMID- 11420982 TI - Complexities of retinal circuitry revealed by neurotransmitter receptor localization. PMID- 11420983 TI - Retinal ganglion cells, glaucoma and neuroprotection. PMID- 11420984 TI - Synaptic inputs to dopaminergic neurons in mammalian retinas. PMID- 11420985 TI - Molecular diversity of gap junctions between horizontal cells. PMID- 11420986 TI - Concepts and challenges in retinal biology. A tribute to John E. Dowling. Introduction. PMID- 11420987 TI - Identification of the first vancomycin-resistant Enterococcus faecalis harbouring vanE in Canada. PMID- 11420988 TI - Outbreak of community-acquired pneumonia caused by Mycoplasma pneumoniae- Colorado, 2000. PMID- 11420989 TI - ECG data compression using wavelets and higher order statistics methods. AB - This paper evaluates the compression performance and characteristics of two wavelet coding compression schemes of electrocardiogram (ECG) signals suitable for real-time telemedical applications. The two proposed methods, namely the optimal zonal wavelet coding (OZWC) method and the wavelet transform higher order statistics-based coding (WHOSC) method, are used to assess the ECG compression issues. The WHOSC method employs higher order statistics (HOS) and uses multirate processing with the autoregressive HOS model technique to provide increasing robustness to the coding scheme. The OZWC algorithm used is based on the optimal wavelet-based zonal coding method developed for the class of discrete "Lipschitizian" signals. Both methodologies were evaluated using the normalized rms error (NRMSE) and the average compression ratio (CR) and bits per sample criteria, applied on abnormal clinical ECG data samples selected from the MIT-BIH database and the Creighton University Cardiac Center database. Simulation results illustrate that both methods can contribute to and enhance the medical data compression performance suitable for a hybrid mobile telemedical system that integrates these algorithmic approaches for real-time ECG data transmission scenarios with high CRs and low NRMSE ratios, especially in low bandwidth mobile systems. PMID- 11420990 TI - A knowledge-based boundary delineation system for contrast ventriculograms. AB - Automated left-ventricle (LV) boundary delineation from contrast ventriculograms has been studied for decades. Unfortunately, no accurate methods have ever been reported. A new knowledge based multistage method to automatically delineate the LV boundary at end diastole (ED) and end systole (ES) is discussed in this paper. It has a mean absolute boundary error or about 2 mm and an associated ejection fraction error of about 6%. The method makes extensive use of knowledge about LV shape and movement. The processing includes a multiimage pixel region classification, shape regression, and rejection classification. The method was trained and cross-validated tested on a database of 375 studies whose ED and ES boundary had been manually traced as the ground truth. The cross-validated results presented in this paper show that the accuracy is close to and slightly above the interobserver variability. PMID- 11420991 TI - Blind separation of slow waves and spikes from gastrointestinal myoelectrical recordings. AB - Myoelectrical recordings of the gut contains slow waves (slow rhythmicity) and spikes (fast rhythmicity). While the slow wave determines the frequency and propagation of gastrointestinal contractions, spike activities are directly associated with the contractions. Traditionally, spikes are extracted from the myoelectrical recording using high-pass/bandpass filters. Due to sharp rising edge (high-frequency component) of the slow wave, the conventional method is not accurate in the separation of the slow wave and spikes, although it has been used for years. In this paper, a novel and fast blind source separation algorithm was developed. Experimental results showed that the proposed method was able to accurately extract spike activities from the myoelectrical recordings obtained in dogs and that its performance was not affected by the high-frequency components of the slow wave. PMID- 11420992 TI - A neural classifier enabling high-throughput topological analysis of lymphocytes in tissue sections. AB - A neural cell detection system (NCDS) for the automatic quantitation of fluorescent lymphocytes in tissue sections is presented in this paper. The system acquires visual knowledge from a set of training cell-image patches selected by a user. The trained system evaluates an image in 2 min calculating: the number, the positions, and the phenotypes of the fluorescent cells. For validation, the NCDS learning performance was tested by cross validation on digitized images of tissue sections obtained from inherently different types of tissue: diagnostic tissue sections across the human tonsil and across an inflammatory lymphocyte infiltrate of the human skeletal muscle. The NCDS detection results were compared with detection results from biomedical experts and were visually evaluated by our most experienced biomedical expert. Although the micrographs were noisy and the fluorescent cells varied in shape and size, the NCDS detected a minimum of 95% of the cells. In contrast, the cellular counts based on visual cell recognition of the experts were inconsistent and largely unreproducible for approximately 80% of the lymphocytes present in a visual field. The data indicate that the NCDS is rapid and delivers highly reproducible results and, therefore, enables high throughput topological screening of lymphocytes in many types of tissue, e.g., as obtained by routine diagnostic biopsy procedures. High-throughput screening with the NCDS provides the platform for the quantitative analysis of the interrelationship between tissue environment, cellular phenotype, and cellular topology. PMID- 11420993 TI - Magnetic resonance image analysis by information theoretic criteria and stochastic site models. AB - Quantitative analysis of magnetic resonance (MR) images is a powerful tool for image-guided diagnosis, monitoring, and intervention. The major tasks involve tissue quantification and image segmentation where both the pixel and context images are considered. To extract clinically useful information from images that might be lacking in prior knowledge, we introduce an unsupervised tissue characterization algorithm that is both statistically principled and patient specific. The method uses adaptive standard finite normal mixture and inhomogeneous Markov random field models, whose parameters are estimated using expectation-maximization and relaxation labeling algorithms under information theoretic criteria. We demonstrate the successful applications of the approach with synthetic data sets and then with real MR brain images. PMID- 11420994 TI - CBIT--context-based image transmission. AB - Few networks offer sufficient bandwidth for the transmission of high resolution two- and three-dimensional medical image sets without incurring significant latency. Traditional compression methods achieve bit-rate reduction based on pixel statistics and ignore visual cues that are important in identifying visually informative regions. This paper describes an approach to managing image transmission in which spatial regions are selected and prioritized for transmission so that visually informative data is received in a timely manner. This context-based image transmission (CBIT) scheme is a lossless form of progressive image transmission (PIT) in which gross structure, represented by an approximate iconic image, is transmitted first. Each part of this iconic image is progressively updated, using a simple set of rules that take into account viewing requirements. CBIT is realized using knowledge about image composition to segment, label, prioritize, and fit geometric models to regions of an image. Tests, using neurological images, show that, with CBIT, a valuable transmitted image is received with a latency that is about one-tenth that of traditional PIT schemes. Frequently, the necessary regions of the image are transmitted in about half the time taken to transmit the full image. PMID- 11420995 TI - Adopting telemedicine services in the airline framework. AB - This paper gives a general overview of the telemedicine service on board airplanes, by considering the problems associated with it, the institutions that are already operating in the field, and the main projects (public and private) that are investigating in this direction. It also reports a brief discussion about the potential market and concludes with a number of issues related to such a service. Most of this information comes from the authors' active participation in several European projects that are particularly focused on telemedicine. PMID- 11420996 TI - Developing a regional healthcare information network. AB - The Internet and associated technology is transforming the dissemination of healthcare information. As this occurs, means must be developed to manage and coordinate it effectively. One approach is through community healthcare information networks (CHINs), which benefit both information providers and consumers. This paper reports on a regional CHIN operational in Scotland. PMID- 11420997 TI - Three-dimensional virtual-reality surgical planning and soft-tissue prediction for orthognathic surgery. AB - Complex maxillofacial malformations continue to present challenges in analysis and correction beyond modern technology. The purpose of this paper is to present a virtual-reality workbench for surgeons to perform virtual orthognathic surgical planning and soft-tissue prediction in three dimensions. A resulting surgical planning system, i.e., three-dimensional virtual-reality surgical-planning and soft-tissue prediction for orthognathic surgery, consists of four major stages: computed tomography (CT) data post-processing and reconstruction, three dimensional (3-D) color facial soft-tissue model generation, virtual surgical planning and simulation, soft-tissue-change preoperative prediction. The surgical planning and simulation are based on a 3-D CT reconstructed bone model, whereas the soft-tissue prediction is based on color texture-mapped and individualized facial soft-tissue model. Our approach is able to provide a quantitative osteotomy-simulated bone model and prediction of postoperative appearance with photorealistic quality. The prediction appearance can be visualized from any arbitrary viewing point using a low-cost personal-computer-based system. This cost-effective solution can be easily adopted in any hospital for daily use. PMID- 11420998 TI - [Certain features of culturing Saccharomyces cerevisiae (Kiev strain) in a wine medium]. AB - The growth of yeast Saccharomyces cerevisiae Kyivsky strain, used in production of champagne, has been studied as affected by a number of factors. The optimum values of saccharose concentration and partial pressure of oxygen dissolved in the medium under which the carbohydrate source use efficiency achieved 9 mln cells/mg of saccharose have been established. The strain growth on the nutrient medium with 2% of saccharose traditionally used in the champagne production has been investigated under the conditions of periodic cultivation. It has been shown that at the beginning of cultivation the low values of the economic coefficient were determined by the Crabtree effect. The saccharose concentration in the culture liquid being decreased to 1%, the carbohydrate utilization efficiency decreases considerably. A chemostatic method of the strain cultivation under growth limitation has been proposed to achieve high concentrations of yeast. Under periodical cultivation the saccharose concentration in the culture liquid would be supported at the level of 0.5-1.0% by its fractional introduction to the fermenter. PMID- 11420999 TI - [Restriction analysis of streptomyces plasmid PSS14]. AB - The plasmid pSS14 having a unique restriction site for endonuclease BamH1 and several identification sites for different restrictases: 5 sites of BglI, 2 sites of BglII, 8 sites of Bsp120I and 3 sites of SacII has been found in the strain Streptomyces sp. 14. Basing on the results of plasmid DNA of pSS14, its molecule weight 13.1 +/- 0.3 tcn has been established. PMID- 11421000 TI - [Ability of representatives of Pantoea agglomerans, as well as Bacillus subtilis and some Pseudomonas species to suppress the development of phytopathgenic bacteria and micromycetes in regulating plant growth]. AB - The ability of representatives of Pantoea agglomerans (Erwinia herbicola (Lohnis) Dye [21]), Bacillus subtilis and some species of Pseudomonas genus to inhibit the growth of phytopathogenic bacteria and micromycetes and to regulate the growth of plants has been comparatively studied. The ability to inhibit the growth of mycellium of phytopathogenic Fusarium avenaceum, F. gibbosum, F. oxysporum was found out in all of 13 investigated strains of P. agglomerans, while the growth of F. culmorum is inhibited by 2 strains and Bipolaris sorokiniana is inhibited by 7 strains. The strains of P. agglomerans and Bacillus subtilis inhibit the growth of mycellium of these mycromycetes to the greater extent than the representatives of Pseudomonas genus. The mycellium growth of B. sorokiniana is better inhibited by B. subtilis and representatives of Pseudomonas genus. Besides the antifungal action 8 strains of P. agglomerans manifested the antagonistic activity in respect to phytopathogenic Agrobacterium tumefaciens and representatives of genera Clavibacter, Erwinia, Pseudomonas, Xanthomonas and also in respect to the microflora which is present in the cabbage and wheat seeds. The strains have been revealed which, parallel with high antagonistic activity in respect to phytopathogenic micromycetes and bacteria, stimulate the seed germination and increase the weight of the cabbage and wheat sprouts. PMID- 11421001 TI - [Prospects for storing streptomycetes in atmospheres of various gases]. AB - The possibility of Streptomyces cultures preservation in the atmosphere of argon, nitrogen, carbon dioxide and free air (the latter as a control) has been first studied. The possibility of short-term (4-6 months) preservation of Streptomyces cultures in the atmosphere of argon and nitrogen has been demonstrated. Some cultures could be preserved in the argon and nitrogen atmosphere for 26 years. PMID- 11421002 TI - [Comparative evaluation of effectiveness of the effect of antibiotics and bacterial substances on clinical trains of Staphylococcus auerus isolated from patients with nonspecific ulcerative colitis]. AB - The article presents of comparative analysis of sensitivity of Staphylococcus aureus clinical strains which were isolated from patients with non-specific ulcerative colitis to 9 widely used antibiotics. Biosporin action against these strains was also studied and a comparison of biosporin and antibiotics actions on the same strain was performed. The high degree of bacilli antagonism to polyresistant S. aureus strains has been revealed. The clear advantage of the therapeutic complex including biosporin as the most effective complex for growth inhibition and elimination of S. aureus from the contents of large intestine has been established. PMID- 11421003 TI - [Increased antibacterial activity of antibiotics with etonium in vitro]. AB - The activity of compositions of antibiotics cefasolin, benzylpenicillin and gentamycin with etonium in respect to museum strains Staphylococcus aureus 209, Escherichia coli K-12, Proteus vulgaris 410, P. mirabilis, Pseudomonas aeruginosa 19, Klebsiella pneumoniae 5054 and polyresistance strains S. aureus, E. coli, P. mirabilis in vitro was researched. The increase of the compositions activity as a result of synergy in the action of their component 4 up to 4496 times has been established. PMID- 11421004 TI - [Baculoviruses--a promising alternative to toxic chemicals in the battle against plant pests]. AB - The economic, ecologic, medical, sanitary and hygienic expediency has been substantiated for the use of Baculoviridae family representatives in creation of preparations for the long-term integrated programs of pest control in natural biocenoses and agrobiocenoses. The program was based on the properties of this unique family. The problems and prospects of biotechnology of baculovirus preparation for pest control in plants in Ukraine have been discussed. PMID- 11421005 TI - [Current methods of laboratory diagnosis of HIV infection]. AB - The paper is a review of scientific data obtained in 1990-1998 and concerning the up-to-date diagnostical approaches used in the laboratory practice. The main widely used methods of HIV-specific antigens and antibodies detection in biological fluids are discussed. The paper deals with the basic methods of HIV infection diagnosis, an analysis of their specificity, sensitivity, their advantages and possible pitfalls. The principal approach permitting one to improve the current test-systems is the elaboration of automated and express tests side by side with the increasing of their sensitivity and specificity. PMID- 11421006 TI - [Functional groups in the alpha-galactosidase active site in Cladosporium cladosporioides]. AB - The activity of alpha-galactosidase isolated from culture fluid of micromycete Cladosporium cladosporioides (Fres.) de Vries 16,038 has been studied as affected by cations, anions and specific chemical reagents (p-chlormercurybenzoate (p ChMB), iodacetamide, N-ethylmaleimide, L-cysteine, dithiotreitol, beta mercaptoethanol, EDTA, o-phenanthroline, sodium azide). It has been established that Ag+ ions inhibited competitively alpha-galactosidase at pH 4.0 and 6.0, the inhibition constants (Ki) made 3.6 x 10(-5) M and 4.3 x 10(-6) M, respectively. Galactose in concentration of 1 mM to 5 mM preserved the enzyme from the negative effect of Ag+ ions, while L-cysteine did not manifest the protective effect. Ions of Hg2+ p-ChMB inhibited noncompetitively the activity of alpha-galactosidase, Ki for Hg2+ and p-ChMB made 5.7 x 10(-7) M and 4.7 x 10(-6) M, respectively. Preincubation with galactose does not preserve alpha-galactosidase from the inhibiting effect of Ag+ and p-ChMB, but th[not readable: see text] compounds (L cysteine, dithiotreitol, beta-mercaptoethanol) restore the enzyme activity. Participation of histidine imidazole group in the catalytic action is supposed on the basis of the inhibitory and kinetic analysis. Sulphydryl groups do not take part in the catalysis but play an important role in supporting the active conformation of the protein molecule. The groups containing the atoms of metals are absent in the alpha-galactosidase molecule. PMID- 11421007 TI - Analysis of predisposing clinical and laboratory findings for the development of endogenous fungal endophthalmitis. A retrospective 12-year study of 79 eyes of 46 patients. AB - PURPOSE: To analyze the predisposing factors for the development of endogenous fungal endophthalmitis (EFE) for its early diagnosis and treatment. SUBJECTS AND METHODS: Seventy-nine eyes of 46 patients with EFE treated in the 12-year period between 1986 and 1998 were included. A retrospective analysis was conducted in respect to age, sex, underlying disease, visual acuity, findings in the anterior and posterior segments, fungal culture of surgical specimens, fever of unknown origin, neutrophils < or = 500/mL, Cand-tec > or = x 4, beta-D-glucan > or = 20 pg, and final visual acuity. RESULTS: The patients were 34 men (74%) and 12 women (26%) between 18 and 78 years of age (mean 57.2 years). Thirty-three of the 46 patients (72%) also were diagnosed with cancer. Fungal infiltration limited to the retina (Stage I) was noted in 13%, budding in the vitreous cavity (Stage II) in 40%, vitreous opacity (Stage III) in 29%, and retinal detachment with Stage III (Stage IV) in 18% of 79 eyes with EFE. Forty patients (87%) were undergoing intravenous hyperalimentation (IVH). The mean interval between the start of IVH and the onset of disease was 11 days. Vitreous surgery was performed in 26 eyes (33%). Candida albicans was detected from surgical specimens in 38%. Fever of unknown origin was noted in 76%, neutrophils < or = 500/mL in 67%, Cand-tec > or = x 4 in 57%, and beta-D-glucan > or = 20 pg in 90% of subjects. CONCLUSION: In patients susceptible to opportunistic infection, beta-D-glucan > or = 20 pg (90%), IVH (87%), fever of unknown origin (76%), male sex (74%), the presence of cancer (72%), neutrophils < or = 500/mL (67%), and Cand-tec > or = x 4 (57%) were considered to be predisposing factors for the development of EFE. PMID- 11421008 TI - Intraocular vancomycin levels after intravitreal injection in post cataract extraction endophthalmitis. AB - PURPOSE: Intravitreal antibiotics are the mainstay of treatment of endophthalmitis following cataract surgery. The purpose of this study is to determine the range of intraocular vancomycin found after intravitreal therapy and assess the optimum time for repeat injections. METHODS: Aqueous and vitreous vancomycin was assayed at the time of reinjection in 14 patients with endophthalmitis showing a poor clinical response after their primary injection. Nine patients received vancomycin 2 mg and another five received vancomycin 1 mg. In six patients the injection was repeated at 48 hours and in seven at 72 hours. Two patients received three injections. RESULTS: Aqueous vancomycin varied from 8.4 to 170 mg/L and the vitreous vancomycin level ranged from 21.2 to 220 mg/L. CONCLUSIONS: In the current study vitreous vancomycin levels were variable, but well within the therapeutic range for sensitive Gram-positive organisms. At times they exceeded the putative retinotoxic levels (100 mg/L). Higher aqueous levels were found after an injection of 2 mg than after 1 mg. Vancomycin levels were still very high 3 days after injection of 2 mg where results were available. Assay at the time of repeat injection may provide insight into the adequacy of vitreous levels and guide future therapy. PMID- 11421009 TI - Erosion and intrusion of silicone rubber scleral buckle. Presentation and management. AB - OBJECTIVE: To describe the clinical presentation and management of erosion and intrusion of silicone rubber implants that are used in scleral buckling procedures for the treatment of retinal detachment. METHODS: The authors identified four patients from their practices during the last 20 years (1978 1998) who had erosion or intrusion of silicone rubber scleral buckles that were used to manage retinal detachment. Approximately 4400 scleral buckling procedures were performed during this period. A retrospective review of the medical records of all patients was performed. Factors that influenced management decisions concerning the intruding buckle are emphasized. RESULTS: All four patients had myopia. The interval between placement of the scleral buckle and development of intrusion ranged from 1 to 20 years. The buckles were intrascleral in three cases and episcleral in one. Recurrent detachment and vitreous hemorrhage were indications for surgical intervention in three cases. After the surgical removal of buckling elements, visual acuity stabilized in all patients and the retina remained attached in all cases. CONCLUSIONS: Erosion and intrusion of scleral buckle are rare complications of scleral buckling procedures. The intruding buckle may be left intact unless there is significant threat to the integrity of ocular structures, recurrent detachment, or hemorrhage. Manipulation of the encircling band or buckle does not necessarily alter the visual acuity or the status of the retina. PMID- 11421010 TI - Vitrectomy for retinal detachments with both peripheral retinal breaks and macular holes. An assessment of outcome and the status of the macular hole. AB - PURPOSE: To establish the effectiveness of vitrectomy and gas tamponade for treating retinal detachments due to peripheral retinal breaks with an associated macular hole and to discover the status of the macular hole at long-term follow up. METHODS: Twenty-three consecutive patients with combined peripheral break and macular hole retinal detachments were treated by pars plana vitrectomy. The main outcome measures were reattachment of the retina and status of the macular hole. RESULTS: Seventy-eight percent of the operations were successful in reattaching the retina initially, improving to 87% after two patients had another operation. Three patients declined further surgery. Long-term follow-up of macular hole status was possible in 16 cases. Closure rate was 31%. CONCLUSION: Pars plana vitrectomy with gas tamponade is an effective method of treating this form of retinal detachment. Some macular holes close after this surgery. PMID- 11421011 TI - Presumed acquired ocular toxoplasmosis in deer hunters. AB - PURPOSE: To describe acquired ocular toxoplasmosis in deer hunters. METHODS AND RESULTS: The authors describe five young men presenting with flu-like symptoms followed by visual loss due to a unilateral, focal necrotizing retinitis. All five men gave a history of ingesting undercooked or uncooked venison. All five had elevated toxoplasma serology, and all five improved clinically with an antitoxoplasma regimen. CONCLUSION: In previously healthy young men, flu-like symptoms associated with visual loss and retinitis should prompt questioning about hunting and raw game meat ingestion, especially when toxoplasmosis is suspected. PMID- 11421012 TI - Indocyanine green angiographic findings in Behcet disease. AB - BACKGROUND: The authors studied indocyanine green (ICG) angiographic features in proven cases of ocular Behcet disease. METHODS: Twenty-six patients (18 male, 8 female; mean age 39.9 +/- 8.9 years) with Behcet disease underwent simultaneous ICG and fluorescein angiography (FA) according to a uveitis angiographic standard protocol. Patients were divided into three groups based on their ocular disease duration: Group A (9 patients), less than 3-year duration; Group B (8 patients), 4- to 10-year duration; Group C (9 patients), more than 10-year duration. The relation between ICG angiographic findings and ocular disease duration and FA signs was delimited. RESULTS: Three findings were disclosed by ICG angiography: 1) poorly defined areas of intermediate and late hyperfluorescence (50% of eyes); 2) well-defined hypofluorescent areas becoming isofluorescent in the late phase (26.92% of eyes); and 3) large, poorly defined hypofluorescent areas visible up to the late phase (30.77% of eyes). The presence of ICG hypofluorescent areas up to the late phase was related to disease duration (P = 0.01), whereas ICG hypofluorescent areas becoming isofluorescent in the late phase were predominant in patients in early stages of ocular disease (P = 0.02). The presence or absence of FA signs did not indicate any significant correlation with the presence or absence of signs revealed by ICG angiography. CONCLUSION: Indocyanine green angiography enabled the identification of different choroidal abnormalities related to the ocular disease duration. The presence of some ICG findings undetectable with FA suggests that ICG and FA are complementary means to diagnose and monitor ocular vascular involvement in patients with Behcet disease. PMID- 11421013 TI - Aging changes of the choroidal dye filling pattern in indocyanine green angiography of normal subjects. AB - PURPOSE: To study the aging changes in the choroid of healthy volunteers with indocyanine green (ICG) angiography. METHODS: Video ICG angiography with adjunctive computer-assisted image analysis was performed on 35 eyes of 30 healthy volunteers (age range, 21-81 years; mean +/- standard deviation, 50.5 +/- 16.2 years) to observe the aging changes of the choroid. RESULTS: In patients in the second and third decades of life, the subfoveal choroidal arterioles fluoresced initially with subsequent rapid filling of the feeding arterioles and choriocapillaris. The watershed zone was clearly observed. In patients older than age 50, the choroidal vasculature filled more slowly. Eventually, the margin of the watershed zone became blurred. The quantitative analysis showed that the number of choroidal arterioles and the fluorescent intensity in the macular region were reduced with age (P < 0.005). In the early venous phase, hypofluorescent patches seen in all ages increased in size and number and remained with aging. The mean fluorescence intensity was not correlated statistically with age. CONCLUSIONS: The features of normal aging patterns of the choroid that we investigated are essential to the interpretation of ICG angiography and may help in understanding the physiologic and pathologic conditions of the choroidal circulations and the choroid and retina themselves. PMID- 11421014 TI - Variations of Weiss's ring. AB - BACKGROUND: Weiss's ring is an important index for diagnosing a posterior vitreous detachment (PVD). The authors studied the configurations of Weiss's ring because the ring has several variations. METHODS: Weiss's ring was examined biomicroscopically in 223 eyes with PVD and documented videographically using a scanning laser ophthalmoscope. The configurations of Weiss's ring were classified into four groups: a complete ring, a partial ring, a ball-like opacity, and a hole without a ring. RESULTS: In the 223 eyes with PVD, Weiss's ring was categorized as a complete ring in 63 eyes (28.3%), a partial ring in 128 eyes (57.4%), a ball-like opacity in 17 eyes (7.6%), and a hole without a ring in 15 eyes (6.7%). A complete ring was observed in 8 (57.1%) of 14 eyes with hyperopia (+3 diopters or more), in 25 (28.7%) of 87 eyes with emmetropia (between -1 and +1 diopter), and in 10 (16.1%) of 62 eyes with myopia (-3 diopters or more). Six months after the initial examinations, the contour of Weiss's ring remained unchanged in 86 (96.6%) of 89 eyes with fresh PVD. CONCLUSIONS: Scanning laser ophthalmoscope is a useful tool for observing Weiss's ring. A "classic" annular opacity around a hole in the prepapillary posterior vitreous cortex represents less than one third of the configurations of Weiss's ring, indicating that the term "ring" is erroneous. PMID- 11421015 TI - Perfluorodecalin-induced intravitreal inflammation. AB - OBJECTIVE: To report an unusual case of intravitreal inflammation in a human eye caused by the presence of residual perfluorodecalin in a case of giant retinal tear and retinal detachment. METHODS: The posterior capsule of the lens, which was infiltrated with deposits, was collected during surgery. The specimen was stained with hematoxylin and eosin, with periodic acid-Schiff, and for melanin. Part of it was examined with electron microscopy. Immunohistochemical staining was performed to demonstrate CD68 antigens, cytokeratin, and glial fibrillary acid protein. RESULTS: Vacuolated macrophages and retinal pigment epithelial cells infiltrated the posterior capsule. Electron microscopy showed the presence of membrane-lined vacuoles within the macrophages. A monolayer of epithelial cells covered the cellular infiltration. CONCLUSION: Residual perfluorodecalin can induce an intraocular chronic macrophage response. PMID- 11421016 TI - Diffuse unilateral hemorrhagic retinopathy associated with accidental perinatal strangulation. A clinicopathologic report. AB - OBJECTIVE: To report an unusual case of diffuse retinal hemorrhage associated with strangulation in a neonate with a tightly wrapped nuchal cord around his neck at birth who was noted to have eyelid and subconjunctival petechial hemorrhages upon delivery. METHODS: Clinical diagnostic examination as well as postmortem gross and histopathologic examination of ocular and central nervous system structures was performed. RESULTS: Funduscopic and gross pathologic examination of the eyes revealed extensive unilateral retinal hemorrhage of the right eye only. This was corroborated by histopathologic studies, which revealed unilateral diffuse hemorrhage throughout all nuclear layers of the retina with a particularly dense nerve fiber layer and sub-internal limiting membrane hemorrhage in the macula along with extraocular muscle and episcleral hemorrhage. Clinical, gross, and microscopic examination also revealed multiple areas of hemorrhage involving the right side of the brain and throughout the cerebellum. CONCLUSION: Although retinal findings in nonaccidental trauma are common, accidental strangulation retinopathy in neonates is a rare occurrence. To the authors' knowledge, this case is the only documented histopathologic study of hemorrhagic retinopathy associated with strangulation in the literature. PMID- 11421017 TI - Diagnostic and therapeutic challenges. PMID- 11421018 TI - Retinal detachment after pneumatic displacement for subfoveal hemorrhage. PMID- 11421019 TI - Retinal venous telangiectasia and anastomoses with cerebellar venous and cavernous malformation. PMID- 11421020 TI - Ultrasound biomicroscopic diagnosis of an occult intrascleral foreign body: an unusual case of ocular siderosis. PMID- 11421021 TI - Combined hamartoma of the sensory retina and retinal pigment epithelium involving the optic disk associated with choroidal neovascularization. PMID- 11421022 TI - Pars plana vitrectomy and endocryocoagulation for paracentral Coats' disease. PMID- 11421023 TI - Pars plana vitrectomy for epiretinal membrane secondary to cat scratch neuroretinitis. PMID- 11421024 TI - Hereditary optic pit and iris coloboma in three generations of a single family. PMID- 11421025 TI - Does silicone oil penetrate the lens capsule? PMID- 11421026 TI - Non-recordable electroretinogram in siderosis bulbi might not indicate poor visual outcome. PMID- 11421027 TI - Chronic postoperative endophthalmitis secondary to Ochrobactrum anthropi. PMID- 11421028 TI - Focal retinal phlebitis as a presenting sign of systemic Bartonella henselae infection. PMID- 11421029 TI - Primary intraocular lymphoma diagnosed by fine needle aspiration biopsy of a subretinal lesion. PMID- 11421030 TI - Are congenital prepapillary arterial loops changeable? PMID- 11421031 TI - An invisible crisis: Pennsylvania's disappearing doctors. PMID- 11421032 TI - In Jeopardy for HCV? PMID- 11421034 TI - Taking the mystery out of health insurance. PMID- 11421033 TI - What did the doctor say? PMID- 11421035 TI - Getting a good nights sleep. PMID- 11421036 TI - Warning: keep your child safe from this summertime danger. PMID- 11421037 TI - Time to clean out that medicine cabinet! PMID- 11421038 TI - Emergency room drama: be prepared before a crisis happens. PMID- 11421039 TI - Emergency contraception and family physicians. An ounce of prevention when it really counts. PMID- 11421040 TI - A tale of two towns. Issues affecting family physicians' choice of practice venue. PMID- 11421041 TI - Another source might explain the pain. PMID- 11421042 TI - Congenital cytomegalovirus infection. Is there a breakthrough? AB - QUESTION: My 26-year-old patient is planning her first pregnancy in the coming month. She works in a day-care centre. Recently, two cases of cytomegalovirus (CMV) infection were diagnosed in her class. What tests should she have before and during the pregnancy, and how should I care for her? ANSWER: Cytomegalovirus infection, the most common congenital viral infection in humans, carries high risk of long-term morbidity and mortality. Seronegative mothers of children in day-care centres are at as high risk of acquiring the infection as day-care workers themselves. The immune status of at-risk patients should be evaluated as pregnancy progresses. Evidence of fetal infection does not necessarily mean fetal disease or damage. With a primary-infected fetus, termination of pregnancy might be discussed with the parents. PMID- 11421043 TI - Cybersearch. Quick clicks to answer clinical questions. PMID- 11421044 TI - Emergency case. Hymenoptera stings. PMID- 11421045 TI - Practice tips. Better safe than sorry. Providing condoms in a family practice. PMID- 11421046 TI - Endometrial sampling for postmenopausal bleeding. Should we put the sampling tools away? PMID- 11421047 TI - What drugs are our frail elderly patients taking? Do drugs they take or fail to take put them at increased risk of interactions and inappropriate medication use? AB - OBJECTIVE: To determine whether there were discrepancies between what medications frail elderly outpatients took and what physicians thought they took and whether discrepancies put patients at risk of taking inappropriate drugs and of increasing the potential for drug interactions. DESIGN: Case series. SETTING: Day Hospital Program at St Mary's of the Lake Hospital in Kingston, Ont. PARTICIPANTS: One hundred twenty community-living elderly patients attending the Day Hospital Program in 1998. Three patients and two family physicians declined to participate. MAIN OUTCOME MEASURES: Lists of medications being taken by patients compared with lists of medications in physicians' charts. Category according to explicit criteria that each drug fell into and risk of drug interactions as determined by the Clinidata Drug Interaction Program. RESULTS: Of the 120 patients, 115 had at least one discrepancy between their lists of medications and their physicians' lists. Of the 1390 medications on the lists, 521 (37%) were being taken by patients without their doctors' knowledge, 82 (6%) were not being taken by patients when doctors thought they were, and 133 (10%) were on both patients' and their doctors' lists but with dosages or frequency of administration that were different. More potential drug interactions were identified on patients' lists than on physicians' lists. No increase in risk of inappropriate drug use was identified. CONCLUSION: Family physicians are often unaware of all the medications their patients are actually taking. Medications used by patients without physicians' knowledge increase the likelihood of drug interactions. Family physicians should look at and inquire about all medications, including over-the-counter drugs, their patients are actually taking. PMID- 11421048 TI - Short report: functional mobility assessment at home. Timed up and go test using three different chairs. PMID- 11421049 TI - [The development of ethics. Identifying what training in medical ethics is needed by family physicians]. AB - OBJECTIVE: To identify what training in medical ethics physician teachers need. DESIGN: Qualitative research study using a modified nominal group technique. SETTING: Family practice units affiliated with the Department of Family Medicine at Laval University in Quebec. PARTICIPANTS: Fifty-three physician teachers in six family practice units. METHOD: During seven meetings, the teachers shared information on clinical situations that had posed ethical problems. Data were analyzed using Strauss and Corbin's method. MAIN OUTCOME FINDINGS: The 277 clinical situations were classified under nine themes: ethics; confidentiality; consent, refusal of treatment, and the right to information; level of care and abstention from and cessation of treatment; relationships with pharmaceutical companies and the ethics of research; ethics of teaching; allocation of resources; influence of third parties; and euthanasia and assisted suicide. Learning objectives were developed. CONCLUSION: This research forms the basis of the ethics curriculum in the family medicine residency program at Laval University. It also offers a strategy for integrating ethics into daily teaching activities because the learning objectives derive directly from the concerns of the teaching faculty. PMID- 11421050 TI - Role of diagnostic labeling in antibiotic prescription. AB - OBJECTIVE: To evaluate the association between diagnostic labeling of respiratory tract infections (RTIs) and antibiotic prescription rates in family practice. DESIGN: Descriptive analysis of outpatient chart review supplemented by interviews with physicians. Charts of patients attending 73 general practitioners were reviewed between October 1997 and February 1998. Two days of practice were evaluated per physician. SETTING: Urban family practices in greater St John's, Nfld. PARTICIPANTS: Of 96 family physicians contacted, 73 (76%) agreed to participate. MAIN OUTCOME MEASURES: Rates of diagnoses and antibiotic prescriptions for acute infections. Physicians were divided into "low prescribers" and "high prescribers" based on overall rates of prescription to patients with infections. Low prescribers were compared with high prescribers with respect to physician characteristics, patient characteristics, and diagnoses assigned. RESULTS: Of all patients seen, 22% were seen for acute infections; RTIs accounted for 76% of diagnoses. Low prescribers and high prescribers were of similar ages and saw similar numbers of patients of similar ages with very similar presenting complaints. Both groups diagnosed urinary tract and skin and soft-tissue infections at similar rates, but differed markedly in their rates of diagnoses of RTIs. High prescribers diagnosed bacterial RTIs in 65.4% (147/225) of their patients; low prescribers diagnosed bacterial RTIs in 31.0% (66/213 (P < .001). CONCLUSION: Family doctors frequently prescribe antibiotics. The difference in rates of prescription between high prescribers and low prescribers is largely explained by assignment of diagnoses of RTIs. PMID- 11421051 TI - Does a third year of emergency medicine training make a difference? Historical cohort study of Queen's University graduates. AB - OBJECTIVE: To describe outcomes of a third-year residency (PGY-3) program in family medicine/emergency medicine in terms of its graduates' practice characteristics and their self-assessed preparedness for practising emergency medicine. DESIGN: A questionnaire was sent to graduates of Queen's University's family medicine residency programs. SETTING: Recent graduates' practices. PARTICIPANTS: All 30 graduates of Queen's University's Family Medicine/Emergency Medicine Program (PGY-3s) from 1988 to 1997 and 90 matched controls chosen randomly from among the 250 graduates of the 2-year family medicine residency program (PGY-2s) during the same period. Six of the 120 were excluded. Response rate was 89%. MAIN OUTCOME MEASURES: Current practice of family and emergency medicine, leadership activities in emergency medicine, self-assessment of preparedness to practise and to lead others at the end of training, self-report of frequency of emergency care situations in subsequent practice for which physicians felt unprepared by their training, and catchment population and "rurality" of location of current practice. RESULTS: Compared with controls, more PGY-3s practiced and took leadership roles in emergency medicine in their hospitals and communities. At the end of their training, PGY-3s reported higher levels of preparedness for practicing and providing leadership in emergency medicine. Both groups reported the same frequency of encountering emergency situations in subsequent practice for which they felt inadequately prepared. Both groups practised in communities of similar size and location. CONCLUSION: Graduates of Queen's University's third-year emergency medicine program appear to practise in accordance with their extra training. PMID- 11421052 TI - Elderly and sun-affected skin. Distinguishing between changes caused by aging and changes caused by habitual exposure to sun. AB - OBJECTIVE: To review and distinguish between skin changes produced by aging and changes produced by habitual exposure to sun. QUALITY OF EVIDENCE: The literature was searched from 1969 to 1999 for articles on dermatoheliosis and sun-damaged skin. Surprisingly few were found comparing the difference between elderly skin and sun-damaged skin. A few articles focused on certain small aspects of sun damaged skin. Many excellent articles described particular changes (e.g., actinic keratosis), but few covered all the changes due to aging and to sun. MAIN MESSAGE: Skin changes due to aging can be distinguished from those due to sun damage. All changes due to sun exposure can be grouped under the term dermatoheliosis; five parts of the skin are involved: epidermis (actinic keratosis), dermis (solar elastosis), blood vessels (telangiectasia), sebaceous glands (solar comedones), and melanocytes (diffuse or mottled brown patches). Habitual exposure to sun and a white skin are prerequisites for developing these changes. Knowing the difference between changes caused by sun and by aging can help physicians predict which patients are most likely to get skin cancers. CONCLUSION: Knowledge of these common skin changes will help physicians diagnose and manage the skin abnormalities of elderly people and of people with dermatoheliosis. PMID- 11421053 TI - Diagnostic challenges in osteoporosis. Indications for bone densitometry and establishing secondary causes. AB - OBJECTIVE: To review indications for assessing bone mineral density (BMD) and to review patient characteristics and diseases associated with osteoporosis. QUALITY OF EVIDENCE: This paper is based on data from longitudinal observational studies of how BMD and other risk factors affect development of fragility fractures and on several peer-reviewed publications describing pathophysiology of bone turnover and pathogenesis of osteoporosis. Indications for obtaining BMD and monitoring treatment are based on the recommendations of the Osteoporosis Society of Canada derived from the consensus opinion of a panel of experts in osteoporosis and based on their review of the primary literature. MAIN MESSAGE: Measurement of BMD provides the best single objective predictor of the relative risk of fracture at sites such as the vertebrae, hip, and wrist, predicting the likelihood of fracture with as much accuracy as measurement of elevated blood pressure predicts stroke. In addition to making the diagnosis of osteoporosis, BMD measurements are used to monitor progression of osteoporosis and effects of therapy. At this date, dual energy x-ray absorptiometry is preferred for measuring BMD. The most likely causes of osteoporosis in any patient are age, hormone withdrawal (in both men and women), and drugs (particularly corticosteroids). Secondary causes, particularly hyperparathyroidism and multiple myeloma, should be excluded by performing appropriate laboratory tests. CONCLUSION: A BMD measurement should be obtained for patients at high risk of osteoporosis and fragility fractures to guide initiation and monitor success of therapy. PMID- 11421054 TI - Case report: sudden painful blindness. Complication of anticoagulant and antiplatelet therapy in patients with disciform macular degeneration. PMID- 11421055 TI - Taking the first steps. Research career program in family medicine. AB - PROBLEM BEING ADDRESSED: Research is not new to family medicine, yet it is pursued less than in other clinical disciplines. We need to establish a critical mass of family medicine researchers. OBJECTIVE OF PROGRAM: To establish a departmental research organization using a strategy implemented in 1995 by the Department of Family and Community Medicine at the University of Toronto. MAIN COMPONENTS OF PROGRAM: We set out to establish a critical mass of researchers. Applicants were required to complete credible and feasible 3- to 5-year research plans and to have formal support from their clinical chiefs. Once selected, researchers were supported for 40% of their time. Support was provided for 3 years and was renewable according to progress on their research plans. Researchers were expected to publish on average two papers yearly and be involved as principal investigator or co-principal investigator on at least one successful grant after the first 3 years. Since implementation in 1996, funded researchers have become principal investigators in 80% of the grants in which they are involved compared with 20% before the support program. Nine of 15 Medical Research Council grants held by family physicians in Canada have department members as principal investigators. Faculty-supported researchers contributed more than 200 peer-reviewed publications to the literature between 1996 and 2000. CONCLUSION: Four years of experience allows for early assessment of the first step taken to build a thriving family medicine research organization using limited departmental resources. PMID- 11421056 TI - Emergency contraception. Summary of the Society of Obstetricians and Gynaecologists of Canada's clinical practice guidelines. PMID- 11421057 TI - New neonatal resuscitation program guidelines. PMID- 11421058 TI - Von Willebrand disease. Little known bleeding disorder. PMID- 11421059 TI - Taking control of your appointment schedule. Part 1: Reconciling income expectations with patient visits. PMID- 11421060 TI - Medical students' career choices. Part 1: Pressure on today's graduates. PMID- 11421061 TI - Evaluation of 1-naphthol as a convenient fluorescent probe for monitoring ethanol induced interdigitation in lipid bilayer membrane. AB - In this work we have tried to evaluate the usefulness of 1-naphthol as an excited state proton transfer fluorescent probe for studying the ethanol-induced interdigitation in lipid bilayer membranes. When ethanol concentration in lipisome is progressively increased, the neutral form fluorescence of 1-naphthol is found to decrease with corresponding increase in the anionic form intensity. This behavior is in contrast to that observed in the absence of lipid where a reverse effect is noticed. Modification of lipid bilayer is known to occur in the presence of ethanol, which increases the packing density of the membrane. Due to this induction of interdigitated gel phase, redistribution of naphthol between the inner core and interfacial region of the lipid bilayer takes places, accounting for the reduction in neutral form fluorescence intensity. The partition coefficient values and the quenching studies also support the redistribution of 1-naphthol in the liposome membrane. The neutral form fluorescence of 1-naphthol successfully monitors the shift in phase transition temperature due to ethanol-induced interdigitation. It also explains the prevention of interdigitation in lipid bilayer at high cholesterol concentration. PMID- 11421062 TI - Gamma and ultraviolet radiation cause DNA crosslinking in the presence of metal ions at high pH. AB - M-DNA is a novel duplex conformation in which metal ions such as Co2+, Ni2+ or Zn2+ replace the imino protons of every base pair. An ethidium fluorescence assay was used to estimate lesions in M-DNA induced by gamma- and UV radiation. General damage to DNA was assessed from the loss of ethidium fluorescence after irradiation of calf thymus DNA. Crosslinks were measured from the return of ethidium fluorescence after a heating and cooling step. Strand breaks were estimated from the loss of fluorescence in covalently closed circular plasmid DNA after a heating and cooling step. For the Co2+ form of M-DNA, gamma-radiation caused the very efficient formation of crosslinks which was not observed with B DNA nor with the Ni2+ or Zn2+ forms of M-DNA. The crosslinks occurred in both A-T and G-C base pairs but did not form in the presence of a free radical scavenger. Crosslinks induced by UV radiation also formed at a faster rate in the Co2+, Ni2+ and Zn2+ forms of M-DNA compared to B-DNA; crosslinking occurred in all DNA but was more prominent in AT-rich sequences and was not inhibited by a free radical scavenger. Therefore, the presence of certain metal ions may lead to large increases in the formation of radiation-induced crosslinks in DNA. PMID- 11421063 TI - Photophysical properties of fluorescent DNA-dyes bound to single- and double stranded DNA in aqueous buffered solution. AB - The absorption and fluorescence spectra, fluorescence quantum yields, lifetimes and time-resolved fluorescence spectra are reported for nine different fluorescent DNA-dyes. The work was initiated in search of a quantitative method to detect the ratio of single-to-double stranded DNA (ssDNA/dsDNA) in solution based on the photophysics of dye-DNA complexes; the result is a comprehensive study providing a vast amount of information for users of DNA strains. The dyes examined were the bisbenzimide or indole-derived stains (Hoechst 33342, Hoechst 33258 and 4',6-diamidino-2-phenylindole), phenanthridinium stains (ethidium bromide and propidium iodide) and cyanine dyes (PicoGreen, YOYO-1 iodide, SYBR Green I and SYBR Gold). All were evaluated under the same experimental conditions in terms of ionic strength, pH and dye-DNA ratio. Among the photophysical properties evaluated only fluorescence lifetimes for the cyanine stilbene dyes allowed a convenient differentiation between ssDNA and dsDNA. The bisbenzimide dyes showed multiexponential decays when bound to either form of DNA, making lifetime-based analysis cumbersome with inherent errors. These dyes also presented biexponential decay when free in aqueous buffered solutions at different pH. A mechanism for their deactivation is proposed based on two different conformers decaying with different kinetics. The phenanthridinium dyes showed monoexponential decays with ssDNA and dsDNA, but there was no discrimination between them. High dye-DNA ratios (e.g. 1:1) resulted in multiexponential decays for cyanine dyes, resulting from energy transfer or self quenching deactivation. Shifts in both absorption and fluorescence maxima for both ssDNA and dsDNA DNA-cyanine dye complexes were small. Broadening of dye ssDNA absorption and fluorescence bands for the cyanine dyes relative to dye dsDNA bands was detected and attributed to higher degrees of rotational freedom in the former. PMID- 11421064 TI - Photosensitization of the sunscreen octyl p-dimethylaminobenzoate by UVA in human melanocytes but not in keratinocytes. AB - Sunscreens penetrate human epidermis and modify the biology of proliferating cells. This study addressed the question whether the UV response of cultured human cells is affected by direct treatment with nontoxic levels of sunscreens. Cell survival following exposure to UVC or unfiltered UBV was not altered by preincubation with 25 micrograms/mL of octyl p-dimethylaminobenzoate (o-PABA), 2 ethylhexyl p-methoxycinnamate (EHMC) or oxybenzone. However, UVA or UVB filtered to reproduce the solar UV spectrum penetrating to the basal layer of the epidermis, highly sensitized cells to killing by o-PABA but not by its hydrolysis product, 4-dimethylaminobenzoic acid. Sensitization was found in all cell types tested, except normal keratinocytes, and could be prevented by certain antioxidants particularly pyruvate and the hydroxyl radical scavenger mannitol. o PABA and EHMC applied without UV reduced the adherence of cells. The results indicate that sunscreens may increase cell mobility and the combination of o-PABA with solar UV may selectively damage melanocytes in the skin. PMID- 11421065 TI - Photostabilizing milled wood lignin with benzotriazoles and hindered nitroxide. AB - The inhibitory processes operating when a 2(-2'-hydroxyphenyl) benzotriazole based ultraviolet absorber (UVA) and a hindered nitroxide free radical are applied to high-yield pulps were studied using milled wood lignin and filter paper as a model. Using quantitative 31P NMR it was determined that the UVA is almost completely photostable during irradiation, suggesting that its protective mechanism is based primarily on ultraviolet absorption. Furthermore, the nitroxide was found to protect the UVA from photodegradation. Analysis of irradiated lignin samples involved derivatization followed by reductive cleavage of beta-arylether groups. The phenolic-OH groups thus released were quantified using 31P NMR. The benzotriazole/nitroxide stabilizing system was found to cooperatively inhibit the cleavage of beta-arylether groups. Furthermore, this system was found to have a synergistic inhibitory effect on the formation of catechol structures. These results suggest that the synergism observed between benzotriazole-based UV screens and nitroxyl radicals on the inhibition of yellowing could be a result of their ability to slow the formation of catechol structures and beta-O-4 cleavage in addition to the possibility of a UVA regeneration mechanism. PMID- 11421066 TI - The light environment and cellular optics of the snow alga Chlamydomonas nivalis (Bauer) Wille. AB - The alga Chlamydomonas nivalis lives in a high-light, cold environment: persistent alpine snowfields. Since the algae in snow receive light from all angles, the photon fluence rate is the critical parameter for photosynthesis, but it is rarely measured. We measured photon irradiance and photon fluence rate in the snow that contained blooms of C. nivalis. On a cloudless day the photon fluence rate at the snow surface was nearly twice the photon irradiance, and it can be many times greater than the photon irradiance when the solar angle is low or the light is diffuse. Beneath the surface the photon fluence rate can be five times the photon irradiance. Photon irradiance and photon fluence rate declined exponentially with depth, approximating the Bouguer-Lambert relationship. We used an integrating sphere to measure the spectral characteristics of a monolayer of cells and microscopic techniques to examine the spectral characteristics of individual cells. Astaxanthin blocked blue light and unknown absorbers blocked UV radiation; the penetration of these wavelengths through whole cells was negligible. We extracted astaxanthin, measured absorbance on a per-cell basis and estimated that the layer of astaxanthin within cells would allow only a small percentage of the blue light to reach the chloroplast, potentially protecting the chloroplast from excessive light. PMID- 11421067 TI - UV doses of Americans. AB - The UV doses of Americans were never measured, but are needed for assessing the risks of UV-related health effects. We calculated these doses using a novel approach. The Environmental Protection Agency's (EPA) National Human Activity Pattern Survey (NHAPS) recorded the activity profiles of 9386 Americans over 24 months to assess their exposure to environmental pollutants, one of which is UV radiation. NHAPS used randomized telephone interviews to get their previous day's minute-by-minute activities. From NHAPS we extracted only the outdoor-daylight data of the northern and southern indoor workers (95%), stratifying by season, sex and age (0-21, 22-40, 41-59 and 60+ years) to find the average time Americans spend outdoors. Knowing the total daylight time and that while outdoors Americans are exposed to about 30% of the available solar UV (on a horizontal plane), we calculated their percent ambients. The average American's percent ambients are 2.6 and 2.5% for northern and southern females, respectively, and 3.5 and 3.6% for northern and southern males, respectively. Men over 40 years of age have the highest ambients (4%). From their ambients we calculated their annual doses using seasonal averages of UV measurements taken daily for over 2 years by EPA Brewer spectrophotometers located in four quadrants of the United States: Atlanta, GA; Boston, MA; Bozeman, MT and Riverside, CA. The average erythemal UV doses of Americans are about 25,000 J/m2/year, 22,000 for females and 28,000 for males, or 33,000 J/m2/year including a conservative continental U.S. vacation (7800 J/m2). Thus, we can now assess the risks of UV-related health effects for Americans. PMID- 11421068 TI - Nanoscale photosynthesis: photocatalytic production of hydrogen by platinized photosystem I reaction centers. AB - A study of the photocatalytic production of molecular hydrogen from platinized photosystem I (PSI) reaction centers is reported. At pH 7 and room temperature metallic platinum was photoprecipitated at the reducing end of PSI according to the reaction, [PtCl6]2- + 4e- + hv-->Pt decreases + 6Cl-, where it interacted with photogenerated PSI electrons and catalyzed the evolution of molecular hydrogen. The reaction mixture included purified spinach PSI reaction centers, sodium ascorbate and spinach plastocyanin. Experimental data on real-time catalytic platinum formation as measured by the onset and rates of hydrogen photoevolution as a function of time are presented. The key objective of the experiments was demonstration of functional nanoscale surface metalization at the reducing end of isolated PSI by substituting negatively charged [PtCl6]2- for negatively charged ferredoxin, the naturally occurring water-soluble electron carrier in photosynthesis. The data are interpreted in terms of electrostatic interactions between [PtCl6]2- and the positively charged surface of psaD, the ferredoxin docking site situated at the stromal interface of the photosynthetic membrane and which is presumably retained in our PSI preparation. A discussion of the rates of hydrogen evolution in terms of the structural components of the various PSI preparations as well as of those of the intact thylakoid membranes is presented. PMID- 11421069 TI - Autofluorescence microscopy of fresh cervical-tissue sections reveals alterations in tissue biochemistry with dysplasia. AB - Fluorescence spectroscopy offers an effective, noninvasive approach to the detection of precancers in multiple organ sites. Clinical studies have demonstrated that fluorescence spectroscopy can provide highly sensitive, specific and cost-effective diagnosis of cervical precancers. However, the underlying biochemical mechanisms responsible for differences in the fluorescence spectra of normal and dysplastic tissue are not fully understood. We designed a study to assess the differences in autofluorescence of normal and dysplastic cervical tissue. Transverse, fresh tissue sections were prepared from colposcopically normal and abnormal biopsies in a 34-patient study. Autofluorescence images were acquired at 380 and 460 nm excitation. Results showed statistically significant increases in epithelial fluorescence intensity (arbitrary units) at 380 nm excitation in dysplastic tissue (106 +/- 39) relative to normal tissue (85 +/- 30). The fluorophore responsible for this increase is possibly reduced nicotinamide adenine dinucleotide. Stromal fluorescence intensities in the dysplastic samples decreased at both 380 nm (102 +/- 34 [dysplasia] vs 151 +/- 44 [normal]) and 460 nm excitation (93 +/- 35 [dysplasia] vs 137 +/- 49 [normal]), wavelengths at which collagen is excited. Decreased redox ratio (17-40% reduction) in dysplastic tissue sections, indicative of increased metabolic activity, was observed in one-third of the paired samples. These results provide valuable insight into the biological basis of the differences in fluorescence of normal and precancerous cervical tissue. PMID- 11421070 TI - Cooperative phenomena in two-pulse, two-color laser photocoagulation of cutaneous blood vessels. AB - A novel laser system has been developed to study the effects of multiple laser pulses of differing wavelengths on cutaneous blood vessels in vivo, using the hamster dorsal skin flap preparation and in vitro, using cuvettes of whole or diluted blood. The system permits sequenced irradiation with well-defined intrapulse spacing at 532 nm, using a long-pulse frequency-doubled Nd:YAG laser, and at 1064 nm, using a long-pulse Nd:YAG laser. Using this system, we have identified a parameter space where two pulses of different wavelengths act in a synergistic manner to effect permanent vessel damage at radiant exposures where the two pulses individually have little or no effect. Using a two-color pump probe technique in vitro, we have identified a phenomenon we call greenlight induced infrared absorption, where a pulse of green light causes photochemical and photothermal modifications to the chemical constituents of blood and results in enhanced infrared absorption. We identify a new chemical species, met hemoglobin, not normally present in healthy human blood but formed during laser photocoagulation which we believe is implicated in the enhanced near-infrared absorption. PMID- 11421071 TI - In vitro induction of PDT resistance in HT29, HT1376 and SK-N-MC cells by various photosensitizers. AB - Our approach to examine the mechanism(s) of action for photodynamic therapy (PDT) has been via the generation of PDT-resistant cell lines. In this study we used three human cell lines, namely, human colon adenocarcinoma (HT29), human bladder carcinoma and human neuroblastoma. The three photosensitizers used were Photofrin, Nile Blue A and aluminum phthalocyanine tetrasulfonate. The protocol for inducing resistance consisted of repeated in vitro photodynamic treatments with a photosensitizer to the 1-10%-survival level followed by regrowth of single surviving colonies. Varying degrees of resistance were observed. The three induced variants of the HT29 cell line were the most extensively studied. Their ratios of increased survival at the LD90 level range between 1.5- and 2.62-fold more resistant. PMID- 11421072 TI - Induction of mRNA for matrix metalloproteinase 1 and tissue inhibitor of metalloproteinases 1 in human skin in vivo by solar simulated radiation. AB - Repeated exposure to solar ultraviolet radiation results in premature skin aging due, in part, to the degradation of dermal collagen by fibroblast collagenase (matrix metalloproteinase 1 [MMP-1]). We have established TaqMan reverse transcription (RT) polymerase chain reaction (PCR) systems to quantify the messenger RNA (mRNA) expression of MMP-1 and its specific inhibitor TIMP-1 in human buttock skin exposed in vivo to solar simulated radiation (SSR). A time course study (n = 6) with two minimal erythema doses (MED) of SSR showed maximal induction of MMP-1 and TIMP-1 at 24 h. A dose-response study (n = 6) sampled at 24 h revealed that doses of about 1 MED were necessary to induce expression of MMP-1 mRNA, and our data suggest that the response is saturated at about 2 MED. We also investigated SSR-induced gene expression in the dermis and epidermis separately (n = 5). MMP-1 was present in both tissues, but TIMP-1 was only detected in the dermis. In general, we could only measure MMP-1 mRNA in the nonirradiated control skin of volunteers who were smokers. We hypothesize very large interpersonal variation with MMP-1 induction compared with TIMP-1 which was detected in all the control sites. This suggests a lack of relationship between MMP-1 and TIMP-1 mRNA expression. The large donor variability for MMP-1 in all the studies demonstrates that it is important to analyze gene expression individually. PMID- 11421073 TI - Long-term effects of photodynamic therapy on fluorescence spectroscopy in the human esophagus. AB - Clinical interest in laser-induced fluorescence (LIF) spectroscopy and photodynamic therapy (PDT) is growing rapidly and may ultimately lead to close parallel use of these techniques. However, variations in LIF due to photosensitizer retention as well as tissue damage and healing processes may interfere with autofluorescence-based diagnostic methods. We have investigated the compatibility of these two techniques by quantifying PDT-induced changes in LIF in the human esophagus. Fluorescence spectra were collected endoscopically at excitation wavelengths (lambda ex) of 337, 400 and 410 nm in 32 patients. Measurements were performed immediately before and after PDT treatment with porfimer sodium and during follow-up procedures. In the months following PDT regions of reepithelialized squamous showed reduced autofluorescence in comparison with untreated squamous regions (P = 0.0007). Photosensitizer fluorescence was undetectable with lambda ex = 337 nm during follow-up procedures, whereas for lambda ex = 400 and 410 nm porfimer sodium fluorescence was noted for nearly a year after treatment. Therefore, residual photosensitizer fluorescence is likely to affect certain LIF-based diagnostic techniques during a period when patients are at high risk for tumor recurrence. Modification of LIF systems and/or the use of alternative photosensitizers may be required to optimize the detection of lesions in the post-PDT patient. Given the potential of LIF as a method for surveillance following cancer therapy, further investigation of the compatibility of specific LIF approaches with cancer pharmaceuticals may be warranted. PMID- 11421074 TI - Monitoring tumor response during photodynamic therapy using near-infrared photon migration spectroscopy. AB - Benzoporphyrin-derivative (BPD)-monoacid-ring A photodynamic therapy (PDT) was performed on subcutaneous tumor implants in a rat ovarian cancer model. In order to assess PDT efficacy the tumor and normal tissue optical properties were measured noninvasively prior to and during PDT using frequency-domain photon migration (FDPM). FDPM data were used to quantify tissue absorption and reduced scattering properties (given by the parameters mu a and mu's, respectively) at four near-infrared (NIR) wavelengths (674, 811, 849 and 956 nm). Tissue physiologic properties, including the in vivo concentration of BPD, deoxy hemoglobin (Hb), oxy-hemoglobin (HbO2), total hemoglobin (TotHb), water (H2O) and percent tissue hemoglobin oxygen saturation (%StO2), were calculated from optical property data. PDT efficacy was also determined from morphometric analysis of tumor necrosis in histologic specimens. All the measured tumor properties changed significantly during PDT. [Hb] increased by 9%, while [HbO2], [TotHb] and %StO2 decreased by 18, 7 and 12%, respectively. Using histologic data we show that long term PDT efficacy is highly correlated to mean BPD concentration in tumor and PDT induced acute changes in [HbO2], [TotHb] and %StO2 (correlation coefficients of 0.829, 0.817 and 0.953, respectively). Overall, our results indicate that NIR FDPM spectroscopy is able to quantify noninvasively and dynamically the PDT induced physiological effects in vivo that are highly correlated with therapeutic efficacy. PMID- 11421075 TI - Effects of ultraviolet-A exposure on ultraviolet-B-induced accumulation of specific flavonoids in Brassica napus. AB - Many plant species are able to acclimate to changes in ultraviolet-B radiation (UVB) (290-320 nm) exposure. Due to the wide range of targets of UVB, plants have evolved diverse repair and protection mechanisms. These include increased biosynthesis of UVB screening compounds, elevated antioxidant activity and increased rates of DNA repair. We have shown previously that Brassica napus L. cv Topas plants can acclimate quite effectively to environmentally relevant increases in UVB through the accumulation of specific flavonoids in the leaf epidermis. However, B. napus was found to lose other flavonoids when plants are exposed to ultraviolet-A radiation (UVA) (320-400 nm) and/or UVB (Wilson et al. [1998] Photochem. Photobiol. 67, 547-553). In this study we demonstrate that the levels of all the extractable flavonoids in the leaves of B. napus plants are decreased in a dose-dependent manner in response to UVA exposure. Additionally, the accumulation of the extractable flavonoids was examined following a shift from photosynthetically active radiation (PAR) + UVA to PAR + UVB to assess if preexposure to UVA affected UVB-induced flavonoid accumulation. UVA preexposures were found to impede UVB-induced accumulation of some flavonoids. This down regulation was particularly evident for quercetin-3-O-sophoroside and quercetin-3 O-sophoroside-7-O-glucoside, which is interesting because quercetins have been demonstrated to be induced by UVB and correlated with UVB tolerance in some plant species. The photobiological nature of these UVA-mediated effects on flavonoid accumulation implies complex interactions between UVA and UVB responses. PMID- 11421076 TI - Photochemically modified alpha-crystallin: a model system for aging in the primate lens. AB - The purpose of this study was to quantitatively study the changes that occur upon irradiation of 3-hydroxykynurenine (3-HK) in the presence of alpha-crystallin under conditions similar to those in the lens. The samples were prepared in 10 mM phosphate buffer at pH 7.4, bubbled with O2 or Ar and irradiated with 300-400 nm light. The amount of light absorbed by the samples (Iabs) was measured using azobenzene as an actinometer. Modifications to alpha-crystallin were monitored by ultraviolet-visible and fluorescence spectroscopy. Aerobic samples had increased absorption around 320 nm and above 400 nm while the 3-HK maximum at 368 nm decreased. The isolated modified protein showed that there was increased absorption throughout the spectrum. Changes in the anaerobic samples were similar to those of the aerobic but occurred more slowly. As irradiation time increased fluorescence emission of the isolated protein red shifted and quantum yields of fluorescence (phi f) were calculated at different irradiation time intervals by comparison to 3-HK. By comparing OD320/OD365 for the model system to values from primate lenses, Iabs can be correlated with age and transmission of the sample in the blue region of the spectrum and thus allows lenticular aging to be quantitated. PMID- 11421077 TI - Recombinant phytochrome A in yeast differs by its spectroscopic and photochemical properties from the major phyA' and is close to the minor phyA": evidence for posttranslational modification of the pigment in plants. AB - Previously, two pools of phytochrome A (phyA' and phyA") have been detected by in situ low-temperature fluorescence spectroscopy and photochemistry; it was suggested that they might differ in the nature of their posttranslational modification. In order to verify this possibility Arabidopsis and rice (Oryza) phyA were expressed in yeast and the pigments were assembled in vivo with phycocyanobilin (PCB) and phytochromobilin (P phi B). The resulting recombinant phytochromes in the red-light-absorbing form (Pr) were characterized in the yeast cell by (1) the fluorescence emission spectra; (2) the temperature dependence of Pr fluorescence intensity and activation energy of fluorescence decay; and (3) the extent of photoconversion of Pr into photoproduct lumi-R (gamma 1) or far-red light absorbing form (Pfr) (gamma 2). Both Arabidopsis phyA/PCB and Oryza phyA/P phi B had low gamma 1 of ca 0.05, allowing their attribution to the Pr" phenomenological type of phytochrome comprising phyA", phyB and cryptogam phytochromes. The spectroscopic properties of Oryza phyA/P phi B were also very close to phyA". However, both investigated holoproteins differed from phyA", both with respect to the character of temperature dependence of the fluorescence yield and activation energy. Thus, recombinant Oryza phyA/P phi B is similar but not identical to phyA". The data demonstrate that the low-abundance-fraction plant phyA (phyA") comes from the same gene as the major (phyA') fraction. Because both endogenous phyA fractions differ from the phytochrome expressed in yeast, they appear to be posttranslationally modified and/or bound to partner proteins or cellular substructures. However, the character of the presumed chemical modification is different in phyA' and phyA" and its extent is more profound in the case of the former. PMID- 11421078 TI - Spectroscopic detection of a phytochrome-like photoreceptor in the myxomycete Physarum polycephalum and the kinetic mechanism for the photocontrol of sporulation by Pfr. AB - Sporulation of the true slime mold Physarum polycephalum (Myxomycetales) can be triggered by the far-red/red reversible Physarum phytochrome. Physarum plasmodia were analyzed with a purpose-built dual-wavelength photometer that is designed for phytochrome measurements. A photoreversible absorbance change at 670 nm was monitored after actinic red (R) and far-red (FR) irradiation of starved plasmodia, confirming the occurrence of a phytochrome-like photoreceptor in Physarum spectroscopically. These signals were not found in growing plasmodia, suggesting the Physarum phytochrome to be synthesized during starvation, which makes the cells competent for the photoinduction of sporulation. The photoconversion rates by R and FR light were similar in the phytochromes of Physarum and etiolated oat shoots. In dark-grown Physarum plasmodia that had not been preexposed to any light only R induced a detectable absorbance change while FR did not. This indicates that most (at least 90%) of the photoreversible pigment occurs in the red-absorbing form. Since the effectiveness of FR in triggering sporulation was enhanced by preirradiation with R, it is concluded that at least part of the Pr can be photoconverted to the active Pfr photoreceptor species. We propose a kinetic mechanism for the photocontrol of sporulation by photoconversion of Pfr, which may also hold for the high irradiance response to FR in Arabidopsis and Cuscuta. PMID- 11421079 TI - Photochemical oxygen consumption, oxygen evolution and spectral changes during UVA irradiation of EMT6 spheroids. AB - Remarkable rates of oxygen consumption are observed via microelectrode measurements immediately upon the onset of 325 nm irradiation of multicell tumor spheroids. Consumption is irradiance dependent over the range 20-200 mW cm-2, and its magnitude is comparable to that observed previously in the same system using exogenous photosensitizers. Oscillations in the oxygen concentrations suggest that oxygen is also being evolved during irradiation. Oxygen evolution is likely the result of enzymatic dissociation of hydrogen peroxide, which is formed through UV-induced photochemistry. Irradiation of spheroids at 442 and at 514 nm produces a much more modest but detectable oxygen consumption. The dynamics of oxygen concentration changes are quite different at these wavelengths, suggesting a different photochemical mechanism. In these cases, initial oxygen depletion is followed immediately by a more gradual, monotonic increase in the oxygen concentration, consistent with irreversible photobleaching. No oscillations in the oxygen concentration are detectable. At 662 nm, no oxygen consumption was observed over the range of irradiances studied. Fluorescence spectra of cells prior to irradiation include contributions from anthranilic acid and reduced nicotinamide adenine dinucleotide (NADH). During 325 nm irradiation, anthranilic acid is rapidly and irreversibly bleached, while NADH emission undergoes only modest reduction. PMID- 11421080 TI - Depth perception in telemedical consultations. AB - This study aimed at quantifying diminished depth perception in telemedicine due to the two-dimensional image and to devise coping strategies for the problem. Two hundred and thirty-five patients in the telemedicine room of a Minor Accident and Treatment Service were studied. The magnitude of impaired depth perception was noted. Seven coping strategies were used and the resolution of the problem was measured. Depth perception was judged to be less than 90% of binocular vision in 235 cases. This improved to more than 90% of binocular vision in 99 of the 234 cases (42.13%) when using of all strategies. Improvement by rotation of the camera 30 degrees at a time in the axial plane was the most useful strategy and it occurred in all 235 (100%) cases. Light adjustment and angulation occurred in 206 of 235 cases (87.66%). Comparison with the opposite side helped in 179 of 235 cases (76.17%), skin color and texture in 139 of 235 cases (59.15%), shutting one eye in 103 of 235 cases (43.83%), enlarging the image in 85 of 235 cases (36.17%), and diminishing depth of field of lens in 77 of 235 cases (32.77%). Other visual cues occurred in 63 of 235 cases (26.81%). Impaired depth perception is a significant problem in telemedicine. It can be improved to make a confident diagnosis in most cases by adopting a variety of strategies that are described in this paper. PMID- 11421081 TI - A telemedicine model for emergency care in a short-term correctional facility. AB - The primary objectives of this research were to determine the effectiveness of a personal computer-based telemedicine system for semi- and nonurgent complaints at a short-term correctional facility and to evaluate the system as a potential model for providing emergency care to remote locations. We performed a retrospective review of medical records of patients referred to the emergency department in person or via telemedicine during a 12-month period. The data included system utilization, chief complaints, physical examination, diagnostic testing, disposition, and outcomes in patients evaluated via telemedicine. Also identified were nursing diagnostic and procedure skills associated with successful evaluation via telemedicine. A total of 530 emergency care records were reviewed with 126 telemedicine consultations performed. Eighty-one of 126 (64%) telemedicine patients remained at the facility following consultation with the remaining 45 (36%) being transported to the emergency department. Rates of return to the emergency department within 7 days following consultation were comparable, patient acceptance and satisfaction was high, and there were no untoward outcomes in the group. Average total time of telemedicine consultation was 30 minutes versus a 2-hour and 45-minutes turnaround time for an emergency department evaluation. A variety of emergency complaints were managed effectively using relatively low-cost computer-based telemedicine technology, thereby eliminating the need for transportation of the patient to the emergency department. This system provides an emergency physician-nurse model for conduction limited emergency care in remote settings. PMID- 11421082 TI - We benefit from fewer OSHA regulations, not more. PMID- 11421083 TI - Sample screen ignored ladder safety. PMID- 11421084 TI - Safety eyewear lens selection. PMID- 11421085 TI - Carpe diem. PMID- 11421086 TI - Posture and power: dynamic use of your body at your computer. PMID- 11421087 TI - A safety double play. PMID- 11421088 TI - QMS for industrial & chemical glove manufacturing. PMID- 11421089 TI - MUTCD's longer reach. PMID- 11421090 TI - Mitigating noise-induced hearing loss. PMID- 11421091 TI - Evolving your safety process. PMID- 11421092 TI - Ergognomics made easy. PMID- 11421093 TI - Emissions control. PMID- 11421094 TI - Solving the work zone puzzle. PMID- 11421095 TI - Safety orientation. PMID- 11421096 TI - Vigilance pays off. PMID- 11421097 TI - Better safety award programs. PMID- 11421098 TI - Lemon juice or lemonade? PMID- 11421099 TI - A solid utility plan. PMID- 11421100 TI - 20 years of safety. PMID- 11421101 TI - Emergency response management. PMID- 11421102 TI - Georgia-Pacific puts safety in prime time. PMID- 11421103 TI - [Why are bone marrow minitransplants being carried out?]. PMID- 11421104 TI - [Atrial flutter. To anticoagulate or not to anticoagulate? That is the question]. PMID- 11421105 TI - Heterozygosity for the H63D mutation in the hereditary hemochromatosis (HFE) gene may lead into severe iron overload in beta-thalassemia minor: observations in a thalassemic kindred. AB - Heterozygosity for beta-thalassemia (minor) by itself does not lead into iron overload; however, when it is inherited together with a homozygous state for either the H63D or the C282Y mutations of the hereditary hemochromatosis gene (HFE gene), iron overload may ensue. We describe here a kindred in which the propositus, being heterozygote for beta-thalassemia and the H63D mutation of the HFE gene, developed severe iron overload and in turn, chronic liver failure with portal hypertension. Other members of the family with either beta-thalassemia or heterozygous for the H63D gene mutation did not develop iron overload. The interaction between beta-thalassemia and hereditary hemochromatosis is briefly discussed and speculations about other possible genetic mutations leading into familial iron loading are done. PMID- 11421106 TI - [Antibiotic therapy in secondary peritonitis: towards a definition of its optimal duration]. AB - BACKGROUND: The optimal time period of antibiotic treatment in secondary peritonitis is still unknown. AIM: To prospectively evaluate: 1) The preferred time period of antibiotic treatment in secondary peritonitis by means of a survey applied to a sample of surgeons, and 2) The outcome of a series of patients with secondary peritonitis in whom the length of antibiotic treatment was based on clinical and laboratory indicators. PATIENTS AND METHODS: A survey presenting the case of a patient with acute appendicitis and secondary peritonitis was applied to 100 Mexican surgeons. The optimal length of antibiotic treatment in that particular patient was asked. Results were tabulated and analyzed. Subsequently, 35 patients with secondary peritonitis were prospectively evaluated. Antibiotics were discontinued when fever relapsed and the white blood count normalized. Clinical characteristics, diagnosis, surgical treatment and outcome were analyzed. Recurrence of sepsis was investigated as the main outcome variable. RESULTS: The survey revealed that 96% surgeons recommended antibiotic treatment for a period longer than one week. In our study group median age was 33 years, 21 were male and 14 female. All patients had a surgical abdominal condition associated to secondary peritonitis. Median period of antibiotic treatment was 3 days. In a follow-up of one month there was no persistent or recurrent sepsis in any patient. CONCLUSIONS: This information may impact in determining the total length of antibiotic therapy if our results are reproduced in studies with more patients. PMID- 11421107 TI - [Enamel hypoplasia in tuberous sclerosis]. AB - Dental enamel hypoplasia is a constant and pathognomonic sign in patients with TS. The defect consists in small cavities (pits) that measure average 80 microns and affect only the enamel without producing lesion to the dentin. We studied the morphology of retained dental pieces of three patients with TS. The pits showed conical and cylindrical shape and measured from 50 to 500 microns. The identification of enamel hypoplasia in deciduous and permanent teeth is of a great value for early diagnosis when other anomalies of TS are not yet produced. PMID- 11421108 TI - [Migration from a rural zone to an urban one is associated with android distribution of body fat in obese women]. AB - INTRODUCTION: Studies about migration to industrialized countries have shown an increased prevalence of diabetes, obesity and dyslipidaemias, all of them related to android body fat distribution. Migration status might be influence body fat distribution but it has not been sufficiently investigated. The aim of this study is to determine the relationship between body fat distribution and migration from rural to urban areas in Mexico. MATERIAL AND METHODS: This sequential sample of 433 women were seen in the outpatient obesity clinic of four federal states: Tabasco (n = 81), Mexico City (n = 166), Coahuila (n = 80), and Yucatan (n = 106). Migration history from rural to urban area, familial history of diabetes, ages of onset of obesity, height and weight circumferences were obtained. A regression logistic model was used and maintained as dependent variable body fat distribution. Age and federal state were considered as confounders and they adjusted the model. RESULTS: Migrating women from rural to urban area were 121 (27.9%). The waist circumference was higher in Tabasco (102.2 +/- 12 cm), and lesser in Yucatan (93.6 +/- 15 cm, p < 0.001); no differences were found for hip circumference. The logistic regression model showed that body fat distribution is associated to migration from rural to urban area, and also to diabetes of mother and age of onset of obesity. CONCLUSIONS: Migrating from rural to urban area is a risk factor for android body fat distribution and this risk increases with age, history of diabetes in mother and adulthood onset o obesity. PMID- 11421109 TI - Absence seizures effects on reading revealed by video-electroencephalography. AB - OBJECTIVE: The effects of absence seizures on reading have not been studied. The purpose of this paper is to show some alterations in reading during absence seizures in patients with video-electroencephalographic recordings. METHODS: Patients were selected if absences seizures were the only type of seizures. They were studied at rest with eyes closed and while reading aloud a text by simultaneous bipolar electroencephalographic and video recordings. RESULTS: Eight patients were studied with 34 events. Absences ranged from 1-12 events in twenty five minutes recording, seizures duration ranged from 1.5-16 seconds. Brief seizures results on non-stop reading, but reading arrest with latency of 1-3 seconds after seizure onset were present in the longer events. Post-absence amnesia ranged from 1-4.5 seconds. Re-start reading occurs mainly in other reading place in the text, but two patients in one event re-started reading in the right place. Classic symptoms during absence were present and reading alterations found were: reading arrest, lost of reading point, bradylexia, dysprosodic reading, grapheme substitution, reading loudness reduction, repeating of phrases. Events occurred without patient seizure awareness, except in one patient by means of the time lost between onset to the end of the event. CONCLUSIONS: This study showed that acute reading alterations during absence seizures are frequent in longer seizures, producing many neuropsychological symptoms that alter reading without seizure awareness. PMID- 11421110 TI - [Homocysteine metabolism and risk of cardiovascular diseases: importance of the nutritional status on folic acid, vitamins B6 and B12]. AB - Homocysteine is a thiol-containing amino acid derived from methionine metabolism that can be degraded through two enzymatic pathways: remethylation and trans sulfuration. In remethylation, homocysteine regenerates methionine. In the trans sulfuration pathway, homocysteine forms cysteine. Due to the rapid metabolic utilization, the plasma concentration of this amino acid is low. Homocysteine circulates as free thiol, homocystine, or bound to free cysteine or to cysteine residues of proteins. Genetic defects of some enzymes in the homocysteine metabolism, or nutritional deficiencies of folic acid, vitamin B6 and B12 lead to an increase in homocysteine plasma concentration and is associated to an increment in cardiovascular diseases. On the basis of clinical and epidemiological studies, homocysteine plasma concentration is considered to be an independent risk factor for the development of atherothrombotic and cardiovascular diseases. The present review describes the homocysteine metabolism, the epidemiological evidence showing the association between homocysteine and the incidence of cardiovascular diseases. The mechanisms by which homocysteine produces vascular damage are indicated. Finally, some recommendations are given for the nutritional therapy of patients with hyperhomocysteinemia. PMID- 11421111 TI - [Hypoalbuminemia in dialysis. Is it a marker for malnutrition or inflammation?]. AB - Hypoalbuminemia in dialysis is a highly prevalent condition associated with morbidity and mortality. Hypoalbuminemia, although not synonymous of malnutrition, is highly related to it. Poor nutrient intake, frequently observed in uremia, may cause malnutrition and subsequently hypoalbuminemia. In addition, it has been recently reported that a systemic inflammatory response may participate in developing hypoalbuminemia in chronic renal failure. Uremia per se, or through mechanisms stimulated by the use of current dialysis membranes and/or solutions, seems to trigger the inflammatory process, remarkably associated with hypoalbuminemia. Infections, to which patients on dialysis are particularly predisposed, stimulate production of the inflammatory response as well. Other conditions non-associated to inflammatory response, such as the protein losses through dialysis, may cause and increase malnutrition. Overhydration, frequently present in patients with renal failure, on the one hand causes dilution of serum albumin concentrations, and on the other hand, is cause of onset and/or enhancement of congestive cardiac failure, which in turn may be associate with malnutrition. Patients with chronic renal failure develop hypoalbuminemia due to a complex setting of conditions, with systemic inflammatory response as a major cause; notwithstanding, other factors such as malnutrition and overhydration can also play a relevant role. Therefore, diagnostic and therapeutic approaches should be individualized. PMID- 11421112 TI - [ANCAS, one, two or three Fates?]. PMID- 11421113 TI - [Zona pellucida antigens in the human ovum: its importance in contraceptive strategies]. AB - The zona pellucida (ZP) is the extracellular matrix surrounding the mammalian oocyte. This matrix consists of three families called ZP1, ZP2 and ZP3. These proteins suffer several posttraductional modifications to give them different immunological and functional properties. In mice has been demonstrated the important role of ZP3 as a receptor of sperm. In the past, research in this field was limited for the difficulty to get enough biological material from different mammalian species, especially from human sources. Recently, several laboratories have expressed ZP recombinant proteins, allowing the study of the proteins under physiologic and pathophysiological conditions, giving the possibility to utilize ZP as a contraceptive target. PMID- 11421114 TI - [Criticism and knowledge]. PMID- 11421115 TI - A case of bilateral seminoma in the setting of persistent mullerian duct syndrome. AB - We are reporting on the case of a 46XY male patient that presented a bilateral testicular seminoma in the setting of bilateral cryptorchidia and left inguinal hernia. The patient received five courses of cisplatin/ciclophosphamide followed by 3,000 cGy of radiation. The patient then, underwent resection of the residual mass and of an unrecognizable pelvic structure. The histopathological analysis revealed necrosis of the residual seminoma and an atrophic uterus. Currently, the patient is alive and asymptomatic at 88 months of follow-up. This is the third case reported of a patient with persistent mullerian duct syndrome and bilateral seminoma. PMID- 11421116 TI - [Genetic effects of acute and chronic ionizing irradiation on Pinus sylvestris L., inhabiting the Chernobyl' meltdown area]. AB - The main results of the 12-year radiation-genetic monitoring of radiobiological, cytogenetic, and genetic parameters in Pinus sylvestris forest plantation from the Chernobyl meltdown area are presented. The acute ionizing irradiation at doses > 1 Gy was shown to induce formation of morphoses and depressed growth; at doses > 2 Gy, the reproductive ability of the trees declined. The radiobiological parameters showed a linear or close to linear dose-dependence relationship. The acute irradiation at a dose of 0.5 Gy induced cytogenetic and genetic effects that were significantly higher than the corresponding control values. The relationship between the cytogenetic effects and the absorbed dose was exponential. The dependence of the mutation frequency at specific loci on the absorbed dose was described by a nonlinear curve. The results of cytogenetic analysis of seedlings obtained from seeds annually collected in zones of slight, moderate, and strong damage of Pinus sylvestris L. are presented. PMID- 11421117 TI - [Chromosomal evolution of the Common Shrew Sorex araneus L.from the Southern Ural and Siberia in the postglacial period]. AB - This paper summarizes a series of studies on chromosomal geography of the common shrew Sorex araneus L. in Siberia and the Southern Urals. Chromosomal races inhabiting the Southern Urals and the Western Siberian Plain sequentially replace each other in the latitudinal direction. In this region, karyotypes of each two adjacent races differ from each other by a single whole-arm reciprocal translocation. In the Eastern Siberian branch, the neighboring races differ mainly in the number or set of metacentric chromosomes. Analysis of the race distribution in the common shrew in the context of paleophysiology of the glacial period allowed us to reconstruct the sequence of events leading to the establishment of the present-day structure of the species. PMID- 11421118 TI - [In silico analysis of the restriction fragments length distribution in the human genome]. AB - The Restriction On Computer (ROC) program (freely available at http://www.mcb.harvard.edu/gilbert/ROC) was developed and used to analyze the restriction fragment length distribution in the human genome. In contrast to other programs searching for restriction sites, ROC simultaneously analyzes several long nucleotide sequences, such as the entire genomes, and in essence simulates electrophoretic analysis of DNA restriction fragments. In addition, this program extracts and analyzes DNA repeats that account for peaks in the restriction fragment length distribution. The ROC analysis data are consistent with the experimental data obtained via in vitro restriction enzyme analysis (taxonomic printing). A difference between the in vitro and in silico results is explained by underrepresentation of tandem DNA repeats in genomic databases. The ROC analysis of individual genome fragments elucidated the nature of several DNA markers, which were earlier revealed by taxonomic printing, and showed that L1 and Alu repeats are nonrandomly distributed in various chromosomes. Another advantage is that the ROC procedure makes it possible to analyze the nonrandom character of a genomic distribution of short DNA sequences. The ROC analysis showed that a low poly(G) frequency is characteristic of the entire human genome, rather than of only coding sequences. The method was proposed for a more complex in silico analysis of the genome. For instance, it is possible to simulate DNA restriction together with blot hybridization and then to analyze the nature of markers revealed. PMID- 11421119 TI - [Molecular genetic analysis of Thrithorax-like gene encoded transcriptional factor GAGA in Drosophila melanogaster]. AB - The Drosophila melanogaster Trithorax-like (Trl) gene is classed with the trx-G genes and codes for several isoforms of the GAGA transcription factor (GAF) which regulates expression of homeotic and numerous other genes. GAF acts as a transcriptional antirepressor, i.e., its interaction with nucleosomal DNA results in the open chromatin conformation in promoter gene regions. The regions thereby become accessible to other transcription factors. As mutations of the Trl gene enhance position effect variegation and disturb chromosome segregation in mitosis and meiosis, GAF is thought to play another, more significant role in determining the chromatin structure. To study the molecular basis of its pleiotropic effect, the Trl gene was subjected to a structural analysis. The genomic Trl gene was sequenced, the sizes of its exons and introns was established, and a complex structure of the 5' and 3' gene regions was demonstrated. The Trl13C, Trl62, DfTrlR67, and DfTrlR85 mutations were exactly mapped. In addition, four insertions of the P element were identified as Trl alleles (Trll(3)s2325, TrlEP(3)3184, TrlEP(3)3191, and TrlEP(3)3609). The viability at various developmental stages was studied in homozygotes for the Trl mutations and in interallelic compounds. The following lethality stages were established: hatching, (Trl13C, DfTrlR85, TrlEP(3)3609), larval molts (Trll(3)s2325), pupation, metamorphosis (DfTrlR67, Trl62), and eclosion (several compounds). PMID- 11421120 TI - [The demonstration of natural hybridization between two swallowtail species Parnassius nomion and Parnassius bremeri (Lepidoptera, Papilionidae) using RAPD PCR technique]. AB - Genetic evidence for interspecific hybridization between Parnassius nomion and Parnassius bremeri in nature is presented. To demonstrate hybridization between these species, RAPD analysis was used. By testing 25 decamer primers, three and two diagnostic markers were revealed for P. nomion and P. bremeri, respectively. Out of 28 animals examined, 4 were shown to be interspecific hybrids. According to the distribution of diagnostic markers, the interspecific hybrids were intermediate with regard to the parental species. Ecological and biological characteristics of two swallowtail species that promote their hybridization in nature are discussed. PMID- 11421121 TI - [Prolongation of MGE 412 transposition induction after gamma-irradiation in an isogenic line of Drosophila melanogaster]]. AB - The dose dependence of the rate of gamma-induced transpositions and consequent dynamics of the MGE 412 pattern after gamma-irradiation were investigated in isogenic line 49 in generations F1, F12, F140, and F170. It was shown that the results on dose dependence of transpositions was very similar with the corresponding results of the classic works by Timofeeff-Ressovsky et al. (1935). It is suggested that the transcribed copies of retrotransposon 412 "cure" gamma radiation-induced double-strand DNA breaks. The phenomenon of prolongation of MGE transposition induction during early generations after treatment was shown. In this period (F1-F12), the maximum transposition rate (lambda approximately equal to 2 x 10(-2) events per MGE copy, per haploid genome, per generation) and the maximum number of heterozygous MGE copies were achieved. In the late generations (F140 and F170), the reduced induction level (lambda approximately 10(-3) was established. In the population of effective size Ne = 2000 individuals, this corresponds to the state when lambda >> 1/4Ne, i.e., when the transposition flow prevails over the MGE copy loss by genetic drift. These data together with some indirect evidence argue for the hypothesis that the spontaneous transposition rate is proportional to the average number of heterozygous MGE copies per diploid genome. PMID- 11421122 TI - [Identification of mutant gene responsible for cotyledons necrosis in developing seedlings of Arabidopsis thaliana]. AB - Genetic and molecular analyses of an Arabidopsis thaliana mutant with necrotic cotyledons from the collection of insertion mutants obtained earlier were conducted. The mutation under study showed incomplete dominance and represented a single insertion of the T region of pLD3 vector used for transformation of germinating seeds to the plant genome during the creation of the collection. Using TAIL-PCR, a fragment of the mutant DNA adjacent to the left border of the T DNA insertion was isolated and sequenced. Computer r-aided analysis showed that the insertion was located on the left arm of chromosome 1. The open reading frame containing the insertion has one exon and encodes a protein of 446 amino acids, whose functions are unknown. PMID- 11421123 TI - [Viral infectious and chromosome aberrations in Bank Vole from natural and laboratory populations]. AB - The frequency of chromosome damage was studied in the carriers of virus of the hemorrhagic fever with renal syndrome (Puumala virus) and in noninfected animals from two laboratory colonies and two natural populations of bank vole. In the laboratory colony, where Puumala virus persisted for three years, multiaberrant ("rogue") cells were found in the bone marrow; the mean frequencies of both structural and numeral chromosome abnormalities were significantly enhanced. In the other laboratory colony, no Puumala virus was detected during all 30 years of its existence, but the mean frequencies of structural chromosome damage were increased to the same degree probably due to the prolonged breeding under laboratory conditions, which resulted in suppression of immunity and DNA repair. The voles from the natural populations were more resistant to the clastogenic viral effect, but they also had multiaberrant cells which served as indicators of viral infection. The data obtained support the hypothesis that viral infections increase mutation rate, contributing thereby to the evolution process. PMID- 11421124 TI - [Polymorphism of cattle prolactin gene: microsatellites, PSR-RFLP]. AB - In the samples of Russian Ayrshire and Gorbatov Red cattle breeds, distribution of frequencies of prolactin (PRL) gene alleles generated due to the presence of polymorphic RsaI site in exon 3 were studied. In the breeds, the frequencies of the B allele of the PRL gene (with RsaI(+) site) detected by the PCR-RFLP method were 14.1 and 8.6%, respectively. In Black Pied, Ayrshire and Gorbatov Red cattle breeds, variation of the microsatellite dinucleotide repeat in the regulatory region of the gene PRL was also studied. Gorbatov Red breed was monomorphic at the microsatellite locus with the only allele 164 bp in length. Two alleles (164 bp and 162 bp) were detected in the other breeds studied. The frequencies of 164 bp allele of the microsatellite locus were 93.7 and 90.0% in Black Pied and Ayrshire breeds, respectively. In Gorbatov Red breed of dairy type with good beef qualities and low milk-fat yield, lower level of heterozygosity for PRL gene was demonstrated compared to Ayrshire and Black Pied breeds with high milk-fat yield. In three cattle breeds, higher mean estimate of polymorphism information content of PCR-RFLP in exon 3 (PIC = 0.21) was revealed compared with the same estimate (PIC = 0.09) for the microsatellite locus variability in the regulatory region of the PRL gene. Characteristics of allele B distribution of the PRL gene in the representatives of the Bovidae family are considered. PMID- 11421125 TI - [Destabilizing effect of chicken selection using the functional adrenal reserves criteria]. AB - Chicken lines produced by divergent selection for the functional adrenal reserves showed significant between-line differences in the content of corticosterone and other hormones (thyroxin, progesterone), as well as in body weight, early maturation, and egg yield. DNA fingerprinting with the pGB725 probe revealed molecular changes in genomic DNA of the chicken lines subjected to plus and minus selection. The genetic distances between the original population and the selected chicken lines, which were estimated from the molecular hybridization patterns, reflected the history of breeding. Analysis of mixed DNA from several individuals of each line revealed specific hybridization bands that could serve as DNA markers during selection for the high and low corticosterone levels in blood. PMID- 11421126 TI - [A2G--a new system of alpha-2-globulin allotypes in pig blood serum]. AB - Combinations of four alpha-2-globulin allotypes were studied for their distribution in pigs of nine different breeds and hybrid groups. Based on this analysis, a new, previously unpublished polyallelic genetic system designated A2G was postulated. The complex alleles of this system control alpha-2-globulin allotypes and are suggested to be encoded by genes of two subloci. One of these subloci is virtually monomorphic, whereas the other has at least four allelic variants. PMID- 11421127 TI - [Genetic association between the apolipoprotein E (ApoE) gene alleles and various forms of Alzheimer's disease]. AB - Allele epsilon 4 of the apolipoprotein E (APOE) gene is associated with higher risk of Alzheimer's disease (AD) in many, though not all, ethnic groups. The APOE allele and genotype frequency distributions were studied in 207 AD patients without cerebrovascular disorders, 62 AD patients with cerebrovascular disorders (combined AD), and 206 control individuals (ethnic Russians from the Russian population). The frequency of allele epsilon 4 in patients with early-onset and late-onset AD was three times higher than in control individuals (p < 0.000001). Compared with control people, patients with cerebrovascular disorders displayed a twofold higher frequency of allele epsilon 4; the difference between the two groups was significant (p = 0.0019). Relative risk of AD in carriers of allele epsilon 4 was five times higher than in carriers of alleles epsilon 2 and epsilon 3 (p < 0.000001). Allele epsilon 2 had a protective effect with respect to AD onset until 65 years of age (p = 0.015). Thus, APOE allele epsilon 4 proved to be a universal factor of early-onset, late-onset, and combined AD in ethnic Russians from Russia. PMID- 11421128 TI - [Characterization of the marriage structure and migration in Chuvash population]. AB - Parameters of the marriage structure and migration were analyzed on the basis of marriage records in the Kanash, Cheboksary, and Morgaushi raions of the Chuvash Republic. Ethnic assortiveness was not detected in Chuvashes and was 4.16-18.05 in Russians. Gene flow between Chuvashes and Russians was 5.1-8.3%; the degree of endogamy was 0.61-0.70. PMID- 11421129 TI - [Polymorphism of the angiotensin-converting enzyme gene in patients with coronary heart disease from Moscow population]. AB - The insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme (ACE) gene was studied in patients with coronary heart disease (CHD) and healthy individuals randomly sampled from the Moscow population. The ACE gene proved to be associated with the plasma apolipoprotein B (ApoB) content in CHD patients, but not associated with HCD development in individuals with elevated serum cholesterol and triglycerides. An association was not revealed between the alleles of the ACE gene and hypertension in CHD patients. PMID- 11421130 TI - [Serotonin type 2a (5-HTR2A) receptor gene polymorphism and personality traits in patients with endogenous psychoses]. AB - Genetic polymorphism of the serotonin receptor (5-HTR2A) gene has been reported to be associated with the expression of clinical signs characteristic of major psychoses, including schizophrenia and affective disorders. In this study, personality traits of patients with these diseases and the associations of these traits with 5-HTR2A allelic polymorphisms were studied. It was demonstrated that schizophrenic and affective patients with the 2/2 genotype of serotonin receptor had lower scores on the anxiety scale and on the anxiety-related hypochondriasis and neuroticism scales than subjects with the 1/1 and 1/2 genotypes. PMID- 11421131 TI - [The database for analysis of quantitative characteristics of chromosome aberration frequencies in the culture of human peripheral blood lymphocytes]. AB - The data on spontaneous chromosome aberration rates in cultures of human peripheral blood lymphocytes obtained in the past 30 years have been collected to form a database. The database contains the results of analysis of more than 330,000 metaphases in lymphocytes from more than 1200 subjects. The frequency of aberrant metaphases in the control group has been estimated at 0.0213 +/- 0.00085. No differences between sexes have been found with respect to either the total chromosome aberration rate or the rates of individual aberration types. The total chromosome aberration rate did not depend on age; however, it has been found that the number of fragments increased and the number of exchanges decreased with age. Smoking has been found to increase the frequency of chromosome aberrations in individuals with occupational hazards, but not in those who are not occupationally exposed to radiation or chemicals. Alcohol consumption increased the frequency of paired fragments, whereas the frequencies of other aberrations did not differ from the control values. PMID- 11421132 TI - [Analysis of Apolipoprotein E gene polymorphism in populations of the Volgo-Ural region]. AB - Polymorphism at the apolipoprotein E gene (ApoE) in populations of the Volga-Ural region was studied by means of polymerase chain reaction. In the region examined the population-specific patterns of the ApoE alleles and genotypes frequency distribution were established. The results obtained were compared with the literature data on the ApoE polymorphism in other world populations. Substantial heterogeneity of different ethnic populations in respect to the ApoE genotypes distribution and frequency was revealed. PMID- 11421133 TI - [Genetic differentiation of the Tuva population with respect to the Alu insertions]. AB - Polymorphism of three rural populations of the Tuva Republic was examined using a set of five autosomal Alu insertions at the ACE, PLAT, PV92, APOA1, and F13B loci. The allele frequency distribution patterns revealed in Tuvinians were typical to Mongoloid populations of Asia and were characterized by relatively high frequency of the Alu-repeat insertion at the PV92 and F13B loci along with relatively low insertion frequency at the APOA1 locus. With respect to the test systems used, Tuvinian populations examined displayed high levels of genetic diversity. The mean expected heterozygosity values in the populations of Kugurtug, Toora-Khem, and Teeli were 0.433, 0.407, and 0.437, respectively. The level of genetic diversity in the pooled Tuvinian sample was 0.432. The coefficient of genetic differentiation in the three populations studied was 1.45 pointing to relatively low level of genetic subdivision of the indigenous Tuvinian populations. However, estimates of genetic differentiation of the Tuvinian gene pool made by use of the Alu-repeat system were higher compared to those performed using classical protein systems, mtDNA, or Y-chromosomal haplotypes. Even though Tuvinian populations were characterized by common gene pool, some features specific to Western Tuvinian population could be distinguished. These features could be associated with higher contribution of the Caucasian component to the gene pool of this population. Phylogenetic analysis demonstrated close genetic relationships between the Tuvinian and Altaic ethnic populations. PMID- 11421134 TI - [Lysine overproduction mutations in the yeast Saccharomyces cerevisiae and its transfection into industrial Yeast strains ]. AB - Yeast mutants resistant to a toxic lysine analog, thialysine were obtained by a method described in the literature. A strain excreting the maximum amount of lysine (0.45 g/l) was selected from these mutants. The intracellular content of lysine was also increased by 30%. The genetic nature of lysine overproduction was studied in this strain. An increase in the amount of excreted lysine was shown to be determined by at least two genes, one of which carries a mutation of thialysine resistance manifesting the pleiotropic effect of lysine overproduction (Th1R) and the other is involved in the regulation of lysine production (PRL). Linkage groups of these genes were determined: the first gene was mapped to the IV chromosome and the second, to the XV chromosome. Both genetic characters were introduced into industrial baker's yeast strains via a series of backcrosses. The stabilization of the genome in the newly derived strains was confirmed by electrokaryotyping. PMID- 11421135 TI - [Production of a SCAR marker in pea Pisum sativum L. using RAPD analysis]. AB - A polymorphic 750-bp fragment, RAPD marker, specific to particular pea genotypes (line L-111 and the Nord cultivar) was identified. Using this RAPD marker, SCAR was obtained. SCAR inheritance in the first and second generations was studied and its dominant character was shown. PMID- 11421136 TI - [Nocturnal street traffic noise and stress hormone excretion in the human]. PMID- 11421137 TI - Finding information in all the wrong places. PMID- 11421138 TI - Prevention and early detection of cancer in the United States. PMID- 11421139 TI - Nurses in all practice settings will benefit from cancer prevention and early detection supplement. PMID- 11421140 TI - Quality of life in patients receiving radiation therapy for non-small cell lung cancer. AB - PURPOSE/OBJECTIVES: To determine whether perceptions of quality of life (QOL) change over time in patients with non-small cell lung cancer (NSCLC) who receive curative radiation therapy (XRT). DESIGN: Descriptive, longitudinal. SETTING: Radiotherapy clinic of a comprehensive cancer center. SAMPLE: 23 patients with NSCLC, selected by nonprobability, consecutive sampling, receiving curative XRT. METHODS: Subjects completed the Functional Assessment of Cancer Therapy-Lung (FACT-L) before, during, and twice after completion of the XRT treatment course. Data were analyzed using descriptive statistics and a multivariate approach to analysis of variance for repeated measures. MAIN RESEARCH VARIABLE: QOL. FINDINGS: FACT-L scores were significantly lower during XRT than before XRT, were significantly higher one month after XRT than before or during XRT, and were not significantly different from the pretreatment level four months after XRT. CONCLUSION: Perceptions of QOL change over time in patients with NSCLC receiving curative XRT. If the study findings are validated in a larger sample, nurses may be able to counsel patients with NSCLC receiving XRT. Nurses can inform patients that although QOL declines during XRT, the change is temporary because QOL will return to a level at least as high as the pretreatment level. IMPLICATIONS FOR NURSING PRACTICE: Nurses need to assess patients perceptions of QOL throughout the course of XRT and assess for sequelae of treatment that affect QOL. Nursing interventions need to be developed and implemented to more effectively manage treatment sequelae and maintain QOL in patients with NSCLC while they receive curative XRT. PMID- 11421141 TI - The value of breast self-examination: meta-analysis of the research literature. AB - PURPOSE/OBJECTIVES: To review 20 studies examining the relationship between breast self-examination (BSE) behaviors, BSE education, the stage of breast cancer at diagnosis, and the mortality or survival rates for breast cancer. DATA SOURCES: 20 studies from medical and nursing journals. DATA SYNTHESIS: In the articles reviewed, the relationship between the value of BSE and BSE behaviors and education (defined as the stage of breast cancer at diagnosis and the mortality or survival rates for breast cancer) was unclear. Methodologic issues such as research design, confounding variables, operational definitions, and sampling were inconsistent and weak. CONCLUSIONS: Meta-analysis suggests that the healthcare research community should state BSE research findings in terms of recommendations for further studies. Until a large number of prospective, randomized, and controlled studies of the relationship between BSE behaviors and education and the value of BSE are conducted, researchers will be unable to make a definitive statement regarding whether BSE provides advantages in detecting breast cancer at an earlier stage, reducing mortality, and increasing survival rates for women with breast cancer. IMPLICATIONS FOR NURSING PRACTICE: Nurse researchers play a key role in conveying the methodologic issues involved in BSE research to the healthcare community and are encouraged to publish their studies in medical journals. When the relationship between the value of BSE and BSE behaviors and education can be confirmed systematically, the healthcare community will recognize and use nurses' research findings to better inform women about breast health. PMID- 11421142 TI - Childhood acute lymphoblastic leukemia: current perspectives. AB - PURPOSE/OBJECTIVES: To provide an overview of childhood acute lymphoblastic leukemia (ALL), including epidemiology, clinical presentation, diagnostic classification, prognostic factors, current treatment, long-term sequelae, and nursing management. DATA SOURCES: Journal articles, books, and clinical experience. DATA SYNTHESIS: Childhood ALL is a heterogeneous disorder, and current treatment is tailored to risk factors (e.g., initial white blood count, cytogenetic properties of the leukemic blasts). Risk-directed therapy ensures that children with a higher risk of relapse receive more intensive treatment, whereas those with lower risk disease receive less toxic therapy with decreased potential for treatment-related morbidity. Quality of life in long-term survivors is a significant issue. Late sequelae of treatment can include neurocognitive difficulties, endocrine dysfunction, secondary malignancies, and cardiomyopathy. CONCLUSIONS: With risk-directed therapy, cure rates for childhood ALL continue to improve. At least 80% of children diagnosed with ALL today are expected to survive their disease. IMPLICATIONS FOR NURSING PRACTICE: Nurses caring for children with ALL can have a significant impact on the children's overall health, from diagnosis through long-term follow-up. Nursing interventions encompass the domains of physical and psychosocial care, as well as patient and family education. Assisting the child and family to maintain normalcy in the face of chronic illness, as well as fostering the family's hope for the future and their belief in the child's potential for survival, are key nursing strategies that promote the child's growth, development, and psychological health. PMID- 11421143 TI - Nurse documentation: not done or worse, done the wrong way--Part II. AB - PURPOSE/OBJECTIVES: To focus on nursing documentation and how it can lead to a malpractice lawsuit. DATA SOURCES: Nursing, non-nursing healthcare and legal journals, case law, and related Internet sources. DATA SYNTHESIS: To avoid liability for inadequate or inaccurate documentation, nurses must be aware of how their documentation can either lead to a malpractice claim or actually decrease their chances of ever being named in a malpractice lawsuit. Malpractice cases often are decided based on documentation. The only viable way to defend against allegations of professional negligence is accurate and complete patient charting or defensive documentation. CONCLUSIONS: By examining case law involving inadequate or inaccurate documentation, nurses will be able to effectively adopt documentation practices or policies to decrease potential litigation. IMPLICATIONS FOR NURSING PRACTICE: Educating nurses about the principles of documentation and the importance of implementing risk-reduction practices will help guard against liability and ultimately improve patient care. PMID- 11421144 TI - Caregiving tasks among family caregivers of patients with lung cancer. AB - PURPOSE/OBJECTIVES: To describe family caregivers' perceptions of time spent and difficulty experienced with performing specific caregiving tasks for patients with lung cancer. To compare adult child and spousal caregivers in relation to time and difficulty of tasks. DESIGN: A secondary analysis of data from a study using a cross-sectional, descriptive, comparison design. SETTING: A university outpatient oncology center, two veterans administration outpatient clinics, and a private outpatient oncology practice. SAMPLE: 78 family caregivers of patients with lung cancer (62 spouses and 16 adult children). METHODS: Data that previously had been collected using a structured interview guide were analyzed using descriptive statistics, comparison of item means, and multivariate analysis of variance. MAIN RESEARCH VARIABLES: Time and difficulty with caregiving tasks. FINDINGS: The most time-consuming tasks for adult children and spouses were providing emotional support, transportation, and monitoring symptoms. The most difficult duties were emotional support, behavioral management, monitoring symptoms, and household tasks. Time and difficulty of tasks did not differ significantly between adult child and spousal caregivers. CONCLUSIONS: Some caregivers may require guidance in providing transportation, emotional support, behavioral management, and symptom management. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses can make a significant impact in assisting family members in providing care by addressing the provision of transportation, emotional support, behavioral management, and symptom management. PMID- 11421145 TI - Music as a therapeutic intervention for anxiety in patients receiving radiation therapy. AB - PURPOSE/OBJECTIVES: To determine whether music moderates the level of anxiety that patients experience during radiation therapy. DESIGN: Experimental, longitudinal, random assignment to music or no music therapy. SETTING: Urban radiation oncology center in a Department of Veterans Affairs hospital in the southeastern United States. SAMPLE: Forty-two men (19 in the experimental group, 23 in the control group) aged 39-80 years (74% white, 12% African American, 12% Hispanic, and 2% other) receiving definitive external beam radiation therapy for pelvic or abdominal malignancies. METHODS: Patients in the experimental group listened to music of their choice provided via audiotapes and headphones before and during their simulation and daily treatments for the duration of the planned course of therapy. The control group received standard care. The State-Trait Anxiety Inventory was administered initially to participants in both groups at the time of evaluation (time 1), post-simulation (time 2), at the end of the first week (time 3), at the end of the third week (time 4), and at the end of the fifth week or end of radiation therapy (time 5). MAIN RESEARCH VARIABLE: State anxiety. FINDINGS: No significant difference existed between the two groups to suggest that music moderated the level of anxiety during radiotherapy. However, post-hoc analyses identified changes and trends in state anxiety scores, suggesting a possible benefit of music therapy during radiotherapy. CONCLUSIONS: Despite a lack of group differences, early intervention with music therapy for patients with high levels of anxiety may be beneficial. IMPLICATIONS FOR NURSING PRACTICE: Nurses and other clinicians may administer state anxiety scales at the initial visit or prior to pretreatment radiation planning (simulation). Individuals who have high state anxiety scores may receive nursing interventions tailored to reduce anxiety during simulation and the early part of radiotherapy. PMID- 11421146 TI - The influence of an organizational pain management policy on nurses' pain management practices. AB - PURPOSE/OBJECTIVES: To examine the influence of a formal organizational pain management policy on nurses' pain management practices. DESIGN: Descriptive correlational. SETTING: Tertiary-care medical center. SAMPLE: 91 nurses providing direct patient care on five study units during a 72-hour study period. METHODS: Off-going nurses completed three self-administered questionnaires one-half hour before the end of each eight-hour work shift. Opioid administration data also were collected. Data were analyzed using correlational, t test, chi-square, and analysis of variance analyses and descriptive statistics. MAIN RESEARCH VARIABLES: Nurses' knowledge of the healthcare organization's chronic pain management policy; nurses' knowledge of pain and pain management; nurses' perceived accountability for pain management activities; the ratio of actual amounts of opioid analgesics administered compared with maximum amounts ordered. FINDINGS: Nurses' knowledge of pain management and their perceived accountability for pain management were significantly related to knowledge of the organization's chronic pain management policy. Correlations were lower than expected, based on theoretical relationships proposed in the open system study framework. CONCLUSIONS: Further research is needed to explore the influence of a high quality pain management policy on nursing practice and clinical pain management. IMPLICATIONS FOR NURSING PRACTICE: Better understanding of the influence of organizational policy could lead to much needed improvements in pain management. PMID- 11421147 TI - Accept me for myself: African American women's issues after breast cancer. AB - PURPOSE/OBJECTIVES: To identify the personal issues and concerns of African American women who are breast cancer survivors. DESIGN: Exploratory. SETTING: Southeastern United States; urban community. SAMPLE: A total of 24 women were recruited from churches and the community; 16 women participated in focus groups. METHODS: Two focus group sessions were held in a community library. Audiotaped interviews were transcribed and analyzed for themes that described issues the women had to deal with after treatment for breast cancer. MAIN RESEARCH VARIABLE: Women's perceptions of the impact breast cancer had placed on their personal lives, including sexuality. FINDINGS: Five themes emerged-body appearance, social support, health activism, menopause, and learning to live with a chronic illness. CONCLUSIONS: African American women have concerns that are similar to, but different from, those of Caucasian women. Further research is needed to identify culturally appropriate care. IMPLICATIONS FOR NURSING PRACTICE: Assess the effects of treatment on women's personal lives. Know where women can purchase prostheses that match their skin tones. Refer minority women to support groups specifically designed for them. PMID- 11421148 TI - SNAP-shots: scenes from nursing action plans--the cultivation of leadership in oncology nursing. AB - PURPOSE/OBJECTIVES: To describe the experiences of oncology nurses who participated in a national workshop to prepare nurses to lead the transformation of cancer care and to illustrate the range of possibilities available to nurses motivated to affect positive change. DATA SOURCES: Literature on leadership and negotiation are referenced and related to the personal experiences and the observations of participants in the workshop. DATA SYNTHESIS: Leadership is presented in historical context. A case study approach was used to organize the personal accounts of participants' plans to affect positive organizational change in oncology care settings. Scenes from action plans illustrate how the workshop's core curriculum, which focuses on leadership models, processes, and skills, was realized in practice. CONCLUSIONS: Leadership is a dynamic process that shapes and is shaped by all stakeholders during the process of planned change. Education, risk taking, and active participation are vital components of the process of leadership development. IMPLICATIONS FOR NURSING PRACTICE: The contributions of nurses may be simple of complex and affect single individuals, groups, or institutions. In all cases, oncology nursing practice and cancer care may be enhanced. PMID- 11421149 TI - Information-seeking behaviors of women with breast cancer. AB - PURPOSE/OBJECTIVES: To explore the information-seeking behaviors of women with breast cancer. DESIGN: Sequential, between-methods triangulation. SETTING: Community-based. SAMPLE: 156 women with breast cancer took part in a mailed survey, and 30 women with breast cancer participated in three focus group discussions (n = 11, 12, and 7). METHODS: Women with breast cancer completed the Miller Behavioral Style Scale (MBSS). Other women with breast cancer participated in one of three age-stratified focus group discussions. MAIN RESEARCH VARIABLE: Information-seeking behaviors of women with breast cancer. FINDINGS: The information-seeking behaviors of women with breast cancer, as measured by the monitoring subscale of MBSS, were not significantly associated with their demographic (e.g., age, socioeconomic class) and illness-related characteristics (e.g., months since diagnosis, surgery, therapeutic regimens). The focus group discussions suggested that the information-seeking behaviors of women with breast cancer were highly individualistic. Although some women actively sought information, others avoided information. In addition, women sometimes fluctuated between seeking and avoiding information. Women sought information to cope with breast cancer, regain a sense of control, increase their feelings of confidence, and help facilitate the decision-making process. Women avoided information to escape from worry, fear, and feelings of negativity and depression. CONCLUSIONS: The information-seeking behaviors of women with breast cancer are highly individualistic and are not related to demographic or disease-related characteristics. IMPLICATIONS FOR NURSING PRACTICE: Breast-care nurses need to be sensitive to the information-seeking behaviors of women with breast cancer. Information seekers should be given maximum information, and information avoiders should be given minimum information. In addition, more research needs to be conducted into the information-seeking behaviors of women with the disease. PMID- 11421150 TI - [Physical exercise and sports: from the best to the worst]. PMID- 11421151 TI - [Hormonal-metabolic adaptations to muscular exercise]. AB - Physical exercise induces numerous metabolic and hormonal changes that may be influenced by various factors, among which the intensity and the duration of the exertion seem to play a major role besides the possible influence of the intake of exogenous substrates. Such modifications are necessary to satisfy the marked increase in energy demand by exercising muscles, while maintaining plasma glucose concentrations within physiological range. The purpose of the present concise review is to analyse the most important fuel-hormonal adaptations observed during short heavy muscular exercise or during prolonged exercise of moderate intensity as well as those observed after physical training in normal subjects. PMID- 11421152 TI - [Sports nutrition]. AB - All nutrition recommendations for athletes must be based on current scientific data and the needs of athletes as individuals. Exercise performance may be improved by adequate food intake. Adequate energy intake needs to be consumed during times of high-intensity training to maintain body weight and maximize the training effect. Carbohydrates are important to maintain blood-glucose and replace muscle glycogen. During exercise, carbohydrates must be provided at the rate of 30 to 60 g per h. Fat intake should not be restricted. No vitamin and mineral supplements should be required if an athlete is consuming adequate energy from a variety of foods. Deshydratation decreases exercise performance. Two hours before exercise 400 to 600 ml of fluid should be consumed and during exercise 150 to 350 ml of fluid every 15 to 20 min. After exercise, the dietary goal is to provide adequate energy and carbohydrate to replace muscle glycogen and ensure rapid recovery. PMID- 11421153 TI - [Physical activity and health in young persons]. AB - Epidemiological studies highlighted an inverse relationship between the level of usual physical activity and the risk of cardiovascular accident. Physical inactivity during childhood and adolescence is believed to be a significant risk factor for various health disorders in later life. One makes a clear distinction between on the one hand, exercise and physical fitness and, on the other hand, activity and health. An investigation into the life style (practice of out-of school physical and sports activities, their frequency, their duration, practice in club and competitive practice) of young Europeans was carried out in nine countries or areas. In the French Community of Belgium, 730 boys and 726 girls of 12 and 15 years were administered an internationally validated questionnaire. The results obtained in all the countries were in agreement with the data of the literature. More than 50% of the girls participated in physical activities only with a very limited frequency and at a low intensity. This is an obvious deficit when compared with the recommendations concerning exercise for health. One can predict without much risk of error that in the adulthood, they will be also classified among the sedentary people. PMID- 11421154 TI - [Children in sports]. AB - A sedentary lifestyle has been linked to the development of cardiovascular disease, diabetes mellitus and obesity. Although these conditions are predominantly found in adults, they are lifelong processes with their origins in childhood. Therefore promotion of physical activity is important in pediatrics. Although some children may be too sedentary, others are participating in training programs and competitive sports that are inappropriate for age. Guidelines for sports participation must be based on a careful consideration of the child's physical fitness, developmental requirements and limitations. PMID- 11421155 TI - [Sports and pregnancy]. AB - Pregnant women consult often their obstetricians for counselling about their way of living. Particularly answering questions concerning physical activity and sports during pregnancy require a profound knowledge on the physiological adaptations of pregnancy and, on the other hand, on performance and sports physiology. On the basis of the current state of research, physical exercise and sport are to be recommended during pregnancy so long as women are aware of potential dangers and contraindications. Maternal benefits and fetal benefits have today been demonstrated. PMID- 11421156 TI - [Multiple benefits of physical exercise in menopausal women]. AB - In postmenopausal women, metabolic and cardiovascular risks are increased not only because of aging but also in relation to estrogen deprivation, decreased physical activity and dietary changes. Accordingly, an increase in total fat mass and its intra-abdominal component, a decrease in lean body mass, an atherogenic dyslipidemia, decreased glucose tolerance, insulin resistance and an increase in procoagulant factors are ensuing, in a manner similar to the metabolic syndrome X and its accompanying risk for cardiovascular disease. Physical exercise concurs with an appropriate diet to weight loss, increases lean body mass and altogether energy expenditure, decreases total body fat mass and visceral fat, improves insulin sensitivity, reduces fibrinogenemia and optimizes hemodynamic parameters. Additionally, physical exercise contributes to protecting bone mass and may be associated with a decreased risk of estrogen-dependent breast and endometrial carcinomas at the postmenopause. In association with a balanced diet, and estrogen administration, physical exercise significantly contributes to metabolic fitness and decreased cardiovascular risk in postmenopausal women. PMID- 11421157 TI - [Physical exercise in the aged]. AB - In the future, physical activities should be more frequent in the elderly population, because of free time and better quality of life. Muscular exercise is very useful; muscular strength and effort ability are preserved in conditioned aged people. It reduces loss of muscle mass, so called sarcopenia; it improves deconditioning and quality of life. In healthy elderly, exercise improves postural status, bone mass and broken risk, cognitive function. However, the impact on falls and immunosenescence is not well known. In ill elderly, quality of life seems better in cardiac and Alzheimer disease, if a control physical activity is performed. Exercise should be adapted and rehabilitation should be done every time it's possible. Social contact and relationships also have good effects, which must be encouraged. PMID- 11421158 TI - [Exercise spirometry in medical practice]. AB - Cardiopulmonary exercise testing is a unique tool to assess the limits and mechanisms of exercise tolerance. It is also useful for establishing the profiles and adequacy of the responses of the systems at submaximal and maximal exercises. The measure of VO2 max (maximal oxygen uptake) has become the "benchmark" to quantify cardiovascular functional capacity and aerobic fitness. PMID- 11421159 TI - [Physiologic evaluation of explosive force in sports]. AB - Although we all possess the capability for anaerobic and aerobic energy metabolism, the capacity for each form of energy transfer varies considerably among individuals. Sports such as football or weightlifting, rely almost exclusively on energy derived from the muscles's pool of high-energy phosphates. Researchers have proposed different tests to estimate the power-generating capacity of high energy phosphates: stair-sprinting power tests, jumping-power tests or sprint cycling. In reality, however, it is difficult to obtain precise physiological or biochemical data during all-out exercise of brief duration. PMID- 11421160 TI - [Beneficial effects of physical activity on cardiovascular risk factors]. AB - Numerous observational (epidemiological surveys) or interventional (controlled trials) studies demonstrated that regular physical activity increases insulin sensitivity in normal subjects and decreases insulin resistance in patients with obesity and/or type 2 diabetes. These favourable effects are at least partially linked to a reduction in abdominal fat mass. This results in a significant improvement of lipid profile, with a decrease in the concentrations of total cholesterol, LDL cholesterol and triglycerides, associated with an increase of HDL cholesterol level. Such favourable metabolic effects related to regular physical activity probably explain the better cardiovascular prognosis observed in regularly exercising subjects as compared to sedentary individuals. These observations should motivate any practitioner to promote endurance physical exercise in every subject, especially in individuals at high cardiovascular risk. PMID- 11421161 TI - [Physical exercise for preventing obesity, promoting weight loss and maintaining weight management]. AB - Obesity is a multifactorial disease making difficult the determination of the precise role of one specific factor such as physical activity or sedentary. Nevertheless, numerous arguments, derived from epidemiological observational studies or from randomized controlled interventional trials, support a favourable role of regular physical activity in the control of body weight. Physical activity contributes to prevent the occurrence of weight excess, especially in children or adolescents, to obtain a satisfactory weight loss, in general in combination with reduced-energy diets, and to maintain long-term weight loss. In addition, physical activity improves both the metabolic profile and health of obese subjects. PMID- 11421162 TI - [Scuba diving: risks for whom and why?]. AB - Scuba diving is an occupation with a still growing expansion. Like every sport it has its own specific pathologies. Some accidents can be very serious and life threatening. We discuss the 3 main groups of accidents: barotraumatism, gas toxicity and accidents related to aquatic environment. The knowledge of the mechanisms underlying these accidents leads to a less risky approach of scuba diving. PMID- 11421163 TI - [Sports under the sun]. AB - The outdoor sports during summer and winter are often performed under uncontrolled exposure to ultraviolet irradiation from sunlight. Dangers are not small for the skin, the eyes and the immune system. Adequate sun protection is recommended. Caution is important in young children. The daily UV index represents a standardized assessment having a regional predictive value for the intensity of the ultraviolet irradiation reaching the biosphere. PMID- 11421164 TI - [The skin, cold and winter sports]. AB - Winter sports are responsible for various dermatoses which could be often avoided by simple preventive procedures. Both the severity and duration of cold exposure combined with wind speed, altitude and environmental hygrometric value govern the potential types of cold injuries. PMID- 11421165 TI - [Skin betrayal, above athletic performance]. AB - Several compounds listed as illicit doping agents can express some effects on the skin. The cutaneous signs are diverse. The clue of the intake of such compounds can be supported by objective non-invasive biometrological assessments. However, such evaluations do not bring the irrefutable proof. The skin can also present unwanted reactions indicating intolerance to the doping agent. Such physiopathological manifestations are not limited to the sport competition, but can also affect some groups of the population searching for a look reminiscent of the ideal young and performing athlete. PMID- 11421166 TI - [Doping in sports]. AB - Doping consists in the use of artificial means or substances with the unique aim of improving performance despite adverse effects on health. Amphetamines stimulate the central nervous system by increasing motivation and vigilance. Often consumed in association with analgesics, they increase the fatigue threshold during prolonged or repeated exercise. Addiction and dependency to these substances are extremely rapid. Side-effects include insomnia, exhaustion, violence and can lead to serious heart diseases. By enhancing capacity for intensive training, anabolic steroids improve strength, alertness and speed. This action is often further strengthened by the use of growth hormones DHEA and IGF 1. Extremely high dosage is used and is in no way comparable with natural secretions or those necessary to re-balance an exhausted glandular system. During prolonged endurance exercise, doping aims at improving the circulation of oxygen in the blood and thus its availability to the muscles. Firstly, the blood haemoglobin concentration was increased by blood transfusions. At present the production of red blood cells is stimulated by repeated injections of exogenous erythropoietin. The extreme viscosity of the blood leads to a risk of vascular thromboses and high blood pressure and accentuates greatly and sometimes even fatally the possibility of brachycardia which is common with sportsmen. PMID- 11421167 TI - [Legal aspects of medicine and sports doping]. AB - Classically, doping is envisaged in terms of the penal or disciplinary consequences it can entail for the sportsman or his (her) sport physician. In our Community, the sportsman who uses doping will in the future not be prosecuted. Another question remains: is a sportsman who was given doping substances by his physician and suffered from this treatment entitled to bring an action against the physician? PMID- 11421168 TI - [Ethical reflections regarding the decree promoting health through sports]. AB - The Parliament of the French Community has edited a decree concerning the promotion of health by sport, the fight against doping in the French Community. This decree declares that federations are responsible for the public health in sport practice. This establishes a link between sport and health. The sport physician must add to his medical practice an ethical dimension. The decree proposes a profound reflection about the good practice in sport medicine. PMID- 11421169 TI - [Pre-competition health screening of young high-level athletes]. AB - The unexpected death of an athlete during the practice of his sport is generally an event largely covered by the newspapers. It raises the problem of screening the athletic population in the aim to disclose anomalies (generally cardiac in origin) that predisposes to sudden death during exercise. In young people, most of these episodes are due to cardiac disorders that can be detected before initiation of the training program. The low prevalence of these disorders, the large number of people to screen and the cost of the examinations however appear to be limiting factors for this type of mass screening. Despite this notion, a scheme of a screening program is proposed in order to prevent as much as possible the occurrence of sudden death in young athletes. PMID- 11421170 TI - [Attitude of general practitioners regarding health certificates for sports participation]. AB - The general practitioner is often asked to produce a certificate indicating an individual's fitness to pratice sport activities. This requires a careful clinical history and physical examination. The opinion of a specialist may at times, be necessary. PMID- 11421171 TI - [Asthma and obesity]. PMID- 11421172 TI - [Comparative study on diagnostic probes: spirometry vs plethysmography in asymptomatic adult asthmatics]. AB - MATERIAL AND METHOD: The study to aim to do the comparison the sensibility between spirometry and pletismography in 20 asthmatic adult free of acute symptoms in basals conditions and to use subsequent of salbutamol spray. RESULTS: Were found to equal value for vital capacity (CV) with p = 0.02 for the both methods. For the total capacity pulmonary (CPT) with p = 0.05, p = 0.02 and increment in residual volume (VA) with p = 0.001 in the pletismography in both conditions (basal and post-salbutamol). The specific conductance (CE) to show p = < 0.001. The flows velocities (VF50, VF25-75) and (VF75, VF75-85) to show similar obstructions for aerial track smaller and smallest with p = < 0.20. CONCLUSION: Than both methods to have the same specificity, but no sensibility. The pletismography apparently to be more sensitive to diagnostic obstruction in airway tract of caliber medium. PMID- 11421173 TI - [Results of an education program for adult asthmatics]. AB - Bronchial asthma is a chronically disease with huge social economical consequences. Educational programs, which have been design for asthmatic patients and diminish asthma mobility and reduce medical cost. They also improve the patients quality of life. We show the results of our educational programs for asthmatic adults which were evaluated through a questionnaire applied to these patient one year after the program was introduced. The results show decreased of medicine required asthmatic crisis. PMID- 11421174 TI - [Clinical diagnostic assay of allergic patients with pollen extract from Parthenium hysterophorous L]. AB - AIMS: There are no antecedents in Cuba of studies developed to prove the efficacy of on allergy test to pollen Parthenium hysterophorus L. OBJECTIVE: Evaluate the allergenic extract pollen Parthenium hysterophorus L elaborated at the reference hospital William Soler City Havana. MATERIAL AND METHODS: We selected 100 patients with a positive clinic history of inhalant allergy and 100 healthy person. All the persons selected were skin prick tested with dillution's of the extract (1:100 and 1:1000 P/V) and positive and negative control. We measured the average diameter of the wheal and the perpendicular diameter, then we calculated the average. We also calculated size of the wheal in the duplicated, which constituted the size of the reaction. We estimated the validity of the test calculating the sensibility and specificity. We used the EPITABLE program to do this. RESULTS: The skin prick tested positive in 79% of the patients and 9% in the healthy persons, for a sensitivity of 79% the interval of confidence (IC), 69.5, 86.2 and specificity of 91%, IC 83.2, 95.5, the predictive positive value 89.8% IC 81.04, 94.9 and the predictive negative value 81.3%, IC 72.5, 87.8. CONCLUSIONS: The allergenic extract of Parthenium hysterophorus L in very sharp to test the allergy to his pollen produced by the allergology laboratory of Hospital William Soler, is efficacy by diagnostic of the alterations caused by this pollen. PMID- 11421175 TI - [Asthma and gastroesophageal reflux. Diagnostic correlation between gammography and digestive endoscopy]. AB - AIMS: Incidence of gastroesophageal reflux (RGE) in asthmatic it is considered be of the rang of 33% until 89%. RGE can be classic or atypical and 30% will be payees of silent reflux. It considers at the moment so much as standard of gold to 24 hours pH esophagus like endoscopic digestive and biopsy, being determined a smaller sensibility to gastric gammagraphic but maybe a bigger utility in relation to their readiness and access easiness. OBJECTIVE: Comparative study is presented that reports sensibility/specificity of gastric gammagraphic, relating it with digestive endoscopic study and biopsy report. METHOD: All asthmatic patient of difficult control and of up to 6 years, selected of the External Consultation of the Service. Gastric gammagraphic was determined and later on the panendoscopic study with taking of biopsy. It integrated sensibility specificity for gammagraphic gastric. RESULTS: Study that understood 8 months, with a total of 44 patients, with a range of 1.4 year-old age to 6 years. Gastric gammagram was reported positive in 25 patients, (56.8%) and negative in 19,(43.18%). Endoscopic studies demonstrated incompetent hiatus in 33 (75%), esophagitis of variable degree in 20 patients, (45.4%), Gastritis in 8 (18.1%) and normal study in 8, (18.1%). Histopathologies: Esophagitis of variable degree in 33 patients (75%), chronic gastritis in 2 patients, (4.5%), Esophagus of Barrett in 2 patients, (4.5%), and normal histopathologic study in 7,(15.9%). Of the 44 patients, 81.5% showed significant reflux. It is integrated a sensibility and specificity of the gastric gammagraphic of 71.4% and 54.2%, respectively. CONCLUSIONS: Gastric gammagraphic contributes sensibility it mediates but acceptable for diagnose of RGE. In asthmatic of difficult control, it should be considered initially endoscopic study. The probability of RGE should always take into account in all asthmatic patient. PMID- 11421176 TI - [Comparative treatment between thalidomide and transfer factor in severe atopic dermatitis]. AB - AIMS: The atopic dermatitis is an chronic inflammatory illness of the skin. It exists an interrelation complex of factors gene, environmental, and psychological that contribute to the development and severity of the illness. The immunol aberrations significant is the answer increased of IgE specific antibodies toward antigens common, the liberation is increased of immunol mediators by the basophils and mast cells, eosinophils peripheral and local, besides enlarges the biphasic activity Th1/Th2 with liberation of cytokines (IL-4, IL-5, IL-13), GM C5F, and decrease of IFN-gamma by the cells Th1. Leung to report a knowledge upon the bases immunopathologies of it atopic dermatitis has immunopathologies clinical important for the diagnosis and processing. Alternatives multiples of processing by the same complexity of the illness exist. OBJECTIVE: To compare the security and the clinical efficacy of the thalidomide and the factor of transfer in the atopic dermatitis severe. MATERIAL AND METHOD: Were studied patient with diagnosis of atopic dermatitis severe in agreement with the criterions of Hanifin and Rajka that they entered to the service of Allergy and Immunology Clinical of the Hospital Regional Lic. Adolfo Lopez Mateos (public hospital). They were included 19 patient (women 12 and men 7, with age average 30 +/- 4 years). They were distributed in two groups. The first group of 5 patient administration thalidomide 200 mg/d during six months. The second group am administered the factor of transfer a total of 15 units by road oral during six months. Studies of laboratory for appraisal were requested immunology and metabolic pretreatment and pretreatment. RESULTS: In the group A dealt with thalidomide 5 patient and the group B dealt with FT, both presented a statistically significant decrease, as for the extension of the wounds (p < 0.01), and 1 am observed greater reduction in the intensity of the symptoms, the SCORAD total (p < 0.001 and p < 0.001 respectively) with statistical difference among them. None presented alterations immunologies and metabolic secondary to the use of the two drugs and not there was the need to suspend the processing. During the period of study, the patient were maintained controlled to the allergic rhinitis and the asthma. DISCUSSION: In the atopic dermatitis by its secondary clinical complexity to the multifactors etiologic, the alternatives of processing utilized in the present study are an option the security and efficacy, I am observed better clinical. PMID- 11421177 TI - HIV surveillance in hard-to-reach populations. PMID- 11421178 TI - HIV surveillance among injecting drug users. AB - Injecting drug users (IDUs) should be considered a 'partially hidden population' at high risk for HIV infection. In almost all locations it should be possible to locate and conduct research with IDUs, but it will probably never be possible to enumerate or draw random samples from an IDU population. Surveillance research studies with IDUs should include risk behaviors, as surveillance of HIV infection only will not be sufficiently time sensitive, and be used to develop and refine HIV prevention programming for the population. Contacts with IDUs can be developed at multiple settings, including voluntary treatment programs, law enforcement settings, and through 'street outreach.' Each type of setting has different advantages, disadvantages and ethical concerns. HIV testing as part of surveillance also raises additional important ethical concerns. The primary risk behaviors that should be included in surveillance studies are 'sharing' of drug injection equipment, the potential for rapid partner change among risk partners, and sexual risk behavior. Additional important objectives for surveillance research include: (1) the size of the local IDU population, (2) patterns of drug use, (3) availability injection equipment, (4) participation in prevention activities, and (5) access to and use of anti-retroviral treatments. HIV incidence is an ultimate objective for surveillance research, but there are no currently available cost-efficient methods for studying HIV incidence, so estimation from indirect measurements is usually required. PMID- 11421179 TI - HIV surveillance among men who have sex with men. AB - OBJECTIVES: To review current approaches to HIV surveillance among men who have sex with men (MSM), identify illustrative best practices and lessons learned, and outline ways to enhance surveillance systems. METHODS: Review of the literature and institutional guidelines for HIV/AIDS, sexually transmitted infection (STI), and behavioral surveillance and summary of results of an international workshop. RESULTS: On-going formative research, HIV/AIDS and STI case reporting, HIV prevalence and incidence studies, and behavioral surveys are essential components of an effective HIV surveillance system for MSM. Alliances with key organizations and actors in MSM communities provide points of access, assist in the development of measures, and guide appropriate use of data. Sampling techniques (convenience, snowball, quota, microsite, time-location, and population-based) offering a range of methods, complexity, and cost have been successfully implemented in MSM communities. Plausible estimates of the size of MSM populations, which are ultimately crucial to the interpretation of surveillance data, can be improved upon using primary and secondary data. CONCLUSIONS: The purpose of HIV surveillance among MSM is not only to monitor disease occurrence and its antecedents, but to regularly use data to plan and evaluate prevention and care programs, advocate for prevention resources, and improve the health, social welfare and human rights of MSM. Practical, incremental steps can be taken to improve HIV surveillance among MSM in all regions of the world in all stages of the epidemic. PMID- 11421180 TI - HIV surveillance among female sex workers. AB - Female sex workers are at high risk for infection with HIV, and their clients may act as a bridging population by spreading HIV to the general population. Comprehensive HIV surveillance among sex workers includes surveillance of HIV infection, of sexually transmitted infections and of risk behavior. Surveillance of HIV infection among sex workers is critical for countries with low-level or concentrated HIV epidemics, and can help in monitoring the response to the HIV epidemic in countries with a generalized epidemic. Sex workers are a vulnerable population, and particular attention needs to be paid to human rights issues including consent, confidentiality and stigma. Collaborations with key players in the local sex work scene--sex workers themselves in the first place--and alliances with salient institutions and groups are key to the success of surveillance among sex workers. Surveillance activities should have a strong link to interventions targeted at sex workers. Surveillance for HIV infection among sex workers can be institution- or community-based. Institutional settings include screening programs for registered sex workers, of sexually transmitted diseases clinics, and re-education camps. Specific sources of bias need to be considered in different settings, and must be measured--through the collection of socio-demographic and behavioral data--to allow a correct interpretation of prevalence data and time trends. Community-based HIV infection surveillance can be conducted in a probability sample of the sex worker population, thereby reducing selection bias. This requires the mapping of sex workers' contact venues, and drawing a random sample from the resulting sampling frame. PMID- 11421181 TI - HIV surveillance in complex emergencies. AB - Many studies have shown a positive association between both migration and temporary expatriation and HIV risk. This association is likely to be similar or even more pronounced for forced migrants. In general, HIV transmission in host migrant or host-forced-migrant interactions depends on the maturity of the HIV epidemic in both the host and the migrant population, the relative seroprevalence of HIV in the host and the migrant population, the prevalence of other sexually transmitted infections (STIs) that may facilitate transmission, and the level of sexual interaction between the two communities. Complex emergencies are the major cause of mass population movement today. In complex emergencies, additional factors such as sexual interaction between forced-migrant populations and the military; sexual violence; increasing commercial sex work; psychological trauma; and disruption of preventive and curative health services may increase the risk for HIV transmission. Despite recent success in preventing HIV infection in stable populations in selected developing countries, internally displaced persons and refugees (or forced migrants) have not been systematically included in HIV surveillance systems, nor consequently in prevention activities. Standard surveillance systems that rely on functioning health services may not provide useful data in many complex emergency settings. Secondary sources can provide some information in these settings. Little attempt has been made, however, to develop innovative HIV surveillance systems in countries affected by complex emergencies. Consequently, data on the HIV epidemic in these countries are scarce and HIV prevention programs are either not implemented or interventions are not effectively targeted. Second generation surveillance methods such as cross sectional, population-based surveys can provide rapid information on HIV, STIs, and sexual behavior. The risks for stigmatization and breaches of confidentiality must be recognized. Surveillance, however, is a key component of HIV and STI prevention services for forced migrants. It is required to define the high risk groups, target interventions, and ultimately decrease HIV and STI transmission within countries facing complex emergencies. It is also required to facilitate regional control of HIV epidemics. PMID- 11421182 TI - Estimating the size of hard-to-reach populations: a novel method using HIV testing data compared to other methods. AB - OBJECTIVE: To estimate population size of hard-to-reach groups such as injecting drug users and men who have sex with men. DESIGN: Several different methods were used to estimate the size of these populations in Canada's three largest cities (Toronto, Montreal and Vancouver). METHODS: A novel method (referred to as the indirect method) was developed for use in Toronto and Vancouver that combines HIV serodiagnostic information with data on HIV testing behavior. Population size estimates were obtained by dividing the number of injecting drug users or men who have sex with men recorded in HIV serodiagnostic databases in a given year by the proportion of the corresponding group that reported being tested in a 1-year period. Results of this method were compared with four other methods: (1) population surveys; (2) capture-recapture (for injecting drug users only); (3) a modified Delphi technique; and (4) a method based on the proportion of never married men aged 45 and over (for men who have sex with men only). Only these other methods were used in Montreal. RESULTS: The survey method gave the lowest estimates which are best viewed as minimum estimates given the relative inability of surveys to access these populations and the reluctance of participants to admit to sensitive behaviors. The indirect method produced results more closely comparable with those obtained by other methods, but they are probably slight overestimates, at least for injecting drug users, due to possible underestimation of the proportion tested for HIV. Point estimates using the indirect method were 17,700 and 17,500 for injecting drug users in Toronto and Vancouver, respectively, and 39,100 and 15,900 for men who have sex with men. In Toronto, results for the other methods ranged from 12,300-13,360 for injecting drug users and 18,800-35,000 for men who have sex with men. For Vancouver, these ranges were 6400-11,670 and 7000-26,500, respectively. In Montreal, ranges were 4300-12,500 for injecting drug users and 18,500-40,000 for men who have sex with men. CONCLUSIONS: This novel method provides estimates of population size of hard-to reach groups such as injecting drug users and men who have sex with men that are comparable with results derived by other methods. These estimates may be useful for the purposes of planning, implementing and evaluating prevention and care services, especially when they are combined with the results of other estimation methods to improve the degree of confidence in the resulting estimates. PMID- 11421183 TI - Laboratory testing and rapid HIV assays: applications for HIV surveillance in hard-to-reach populations. AB - Most HIV surveillance has been performed through serologic surveys in relatively stable, accessible populations. Similar surveillance, with or without counseling and testing, in populations that are hard-to-reach, presents logistical challenges, including the selection of laboratory testing strategy and algorithm. The advent of rapid serologic assays for HIV now allows for on-site testing, including confirmatory testing, and rapid provision of test results and counseling. The possibility of only a single contact makes repeat sampling, which current diagnostic testing recommendations include, difficult. To address the logistical complexities in surveillance in hard-to-reach populations and the increased availability of rapid tests, we propose adapting the testing strategies for HIV of the World Health Organization/the joint United Nations Programme on HIV/AIDS in order to facilitate this surveillance, including, where carried out, the provision of test results back to individuals. The choice of enzyme-linked immunosorbent assay (ELISA) versus rapid testing for these settings is discussed, as is the choice of specimen--blood, oral fluid, or urine. Three appendices summarize: (1) test algorithms for the various testing strategies; (2) advantages and disadvantages of ELISA and of rapid test formats, and (3) the characteristics and status of currently available rapid HIV tests. We also discuss the potential application of the recently developed 'detuned' methodology for estimating HIV incidence in hard-to-reach populations. PMID- 11421184 TI - [The electronic medical record]. PMID- 11421185 TI - [Driver's licence to patients with heart disease]. PMID- 11421186 TI - [Multidisciplinary electronic medical record and clinical database all in one. SCIBase]. PMID- 11421187 TI - [Improved treatment of diabetic foot problems]. PMID- 11421188 TI - [Epigenetic modification of the genetic material. Genomic imprinting and its significance for disease in human beings]. AB - Genomic imprinting is the epigenetic differential marking of maternally and paternally inherited alleles of specific genes or chromosomal subregions during gametogenesis, leading after fertilization to differential expression during development. Expression is thus monoallelic, with one parental allele being expressed, the other silenced. Imprinting implies the existence of a reversible imprinting signal, which is erased in the gonads to be reset according to the sex of the individual. Mutations in imprinted genes are not inherited in a regular Mendelian fashion. The number of identified imprinted genes is now around 35. Three congenital human disorders are known to be caused by errors in the expression pattern of imprinted genes: Prader-Willi syndrome, Angelman syndrome and Beckwith-Wiedemann syndrome. A number of cancers are also caused by errors in imprinted genes. PMID- 11421189 TI - [The effect of supervised rehabilitation on ankle joint function and the risk of recurrence after acute ankle distortion]. AB - INTRODUCTION: The effect of an early rehabilitation programme, including postural training, on ankle joint function after an ankle ligament sprain was investigated prospectively. METHODS: Ninety-two subjects, matched for age, sex, and level of sports activity, were randomised to a control or training group. All subjects received the same standard information about early ankle mobilisation. In addition, the training group participated in supervised physical therapy rehabilitation (one hour, twice weekly) with emphasis on balance training. Postural sway, position sense, and isometric ankle strength were measured six weeks and four months after the injury, and at 12 months data on re-injury were collected. RESULTS: In both the training group and the control group, there were a significant difference between the injured and the uninjured side for all variables except for position sense at six weeks. The side-to-side differences in per cent were similar for both groups for all variables (p > 0.05) at six weeks, and there were no such differences at four months. Re-injury occurred in 11/38 (29%) is the control group, but in only 2/29 (7%) in the training group (p < 0.05). CONCLUSION: These data showed that an ankle injury led to reduced ankle strength and postural control at six weeks, but that these variables had become normal at four months, irrespective of supervised rehabilitation. However, the findings also showed that supervised rehabilitation may reduce the number of re injuries, and may therefore play a role in injury prevention. PMID- 11421190 TI - [Focus group interview. A method of study and implementation]. AB - INTRODUCTION: The aim of this article is to present the qualitative focus group interview as a useful method of evaluating psycho- and milieu therapeutic treatment. MATERIAL AND METHODS: We conducted two focus group interviews with former inpatients of a psychiatric ward specialising in group therapy. To enhance the quality of the data by triangulation, the staff, representing both milieu- and psychotherapists, were also interviewed. RESULTS: Analysis of the results revealed the following dominant themes: The continuation of the treatment was jeopardised by the existence of a welcome group. There was a need for further information as soon as the patient came into contact with the hospital. Moreover, an earlier and increased involvement of the family was required. After their own interview, the staff participated in deciding which results should lead to alterations in treatment procedures, thereby becoming involved in implementing the results. DISCUSSION: The focus group interview is a valuable method of evaluating psycho- and milieu therapeutic treatment. Interviewing the staff served as triangulation and eased the implementation of the results remarkably. PMID- 11421191 TI - [Osteomyelitis. A rare and serious complication of inguinal hernia surgery]. AB - We report two cases of osteomyelitis following herniotomy ad modum Lichtenstein with implantation of a polypropylene mesh. With the new prosthetic materials, the hernia recurrence rate and sick leave are reduced. In the case of persistent wound infection serious complications might occur. PMID- 11421192 TI - [Picture of the month: Peritonsillar abscess]. PMID- 11421193 TI - [Current psychiatry and modern depression]. PMID- 11421194 TI - [Psychological aspects of nutritional therapy]. PMID- 11421195 TI - [Regeneration of post-infarction heart muscle with bone marrow stem cells]. PMID- 11421196 TI - [Om trans fatty acids from cow's stomach]. PMID- 11421197 TI - [Does DAMP belong in child and adolescent psychiatry?]. PMID- 11421198 TI - [The eight decade--heart massage-no, thank you]. PMID- 11421199 TI - Web alert. PMID- 11421200 TI - Knockouts and transgenics confirm the importance of adrenomedullin in the vasculature. PMID- 11421201 TI - Neuropeptide FF: new molecular insights. PMID- 11421202 TI - Non-surmountable antagonism: a general drawback of pre-steady-state measurement? PMID- 11421203 TI - Visual vignette. Albright's hereditary osteodystrophy(AHO). PMID- 11421204 TI - [Drug transporter: Molecular mechanism of polyspecific substrate recognition]. PMID- 11421205 TI - Coinheritance of factor V Leiden (F5 1691A) and prothrombin 20210 A (F2 20210A) in an Austrian family as cause for early onset of venous thromboembolism. PMID- 11421206 TI - Drug therapy for management of obesity. PMID- 11421207 TI - Drug therapy for management of obesity. PMID- 11421208 TI - Drug-resistant tuberculosis. PMID- 11421209 TI - Efficacy of atovaquone-proguanil? PMID- 11421210 TI - Ethics of research. PMID- 11421211 TI - Omeprazole, its isomers, and the carcinoid question. PMID- 11421212 TI - Tamoxifen's effect in women with breast cancer. PMID- 11421213 TI - Tamoxifen's effect in women with breast cancer. PMID- 11421214 TI - Bladder dysfunction in infants. PMID- 11421215 TI - Nutritional status in children. PMID- 11421216 TI - Conn's syndrome and atrial fibrillation. PMID- 11421217 TI - Targeting voltage-gated Ca2+ channels. PMID- 11421218 TI - Antenatal antibody screening. PMID- 11421219 TI - Ciprofloxacin resistance in Neisseria gonorrhoeae. PMID- 11421220 TI - Squirrel monkey (Saimiri spp.) research and resources. PMID- 11421221 TI - Squirrel monkey behavior in research. PMID- 11421222 TI - Role of the squirrel monkey in parasitic disease research. PMID- 11421223 TI - Pneumococcal conjugate vaccines in Europe, Berlin, Germany, 23-25 August 2000. Report of a European advisory board meeting. PMID- 11421224 TI - Bibliography. Current world literature. Lipid metabolism. PMID- 11421225 TI - Clinical recommendations for oxcarbazepine. AB - Oxcarbazepine (OXC) is indicated for treating partial-onset with or without secondary generalized tonic-clonic seizures, in both adults and children aged over 6 years, as monotherapy or adjuvant therapy. Trials data and extensive clinical experience demonstrate generally good tolerability and antiepileptic efficacy similar to carbamazepine (CBZ), sodium valproate or phenytoin. Since the UK launch of OXC in March 2000, UK collaborators have pooled experience to optimize prescribing recommendations for adults. Many patients are successfully managed using the prescribing information recommended titration schedule. However, evolving clinical experience suggests a slower introduction is preferable (e.g. 150 mg day one, then 300 mg daily, increased by 300 mg weekly) both for monotherapy and adjuvant therapy. Overnight 'switch' from CBZ to OXC (using CBZ:OXC ratio of 1:1.5) has been used for patients responsive to CBZ, but with dose-related side-effects. Owing to individual variations in CBZ enzyme autoinduction, however, overnight switching is advised only for those on CBZ < 800 mg daily; otherwise, slower switching is recommended. OXC is not the first choice alternative for patients developing a CBZ rash owing to increased OXC rash rate in CBZ sensitive subjects. Hyponatraemia may be more common (albeit often asymptomatic) than trials data suggest, especially in the elderly. Serum sodium monitoring is unnecessary, however, unless relevant risk factors or pointers exist. Severe haematological dyscrasias have not been reported with OXC. The enzyme inducing interaction of OXC with ethinyloestradiol and levonorgestrel necessitates additional precautions for women using hormonal contraception. PMID- 11421227 TI - Higher incidence of diabetic nephropathy in type 2 than in type 1 diabetes in early-onset diabetes in Japan. PMID- 11421228 TI - ACE inhibition or angiotensin receptor blockade: impact on potassium in renal failure. PMID- 11421229 TI - Health outcomes associated with calcium antagonists. PMID- 11421230 TI - Steroids in the nervous system: a Pandora's box? PMID- 11421231 TI - 'Saccadic suppression'- no need for an active extra-retinal mechanism. PMID- 11421232 TI - BDNF and epilepsy--the bad could turn out to be good. PMID- 11421233 TI - On the amount of prismatic correction in cases of angular ametropia and diplopia: a preliminary communication. PMID- 11421234 TI - Prothrombin 20210A-associated thrombosis may need concurrency of another prothrombotic factor. PMID- 11421235 TI - [Orthodontics online]. PMID- 11421236 TI - [Research center for dental products and materials]. PMID- 11421237 TI - [Question of the month: presence of spirochetes?]. PMID- 11421238 TI - The case of the missing record (and its consequences). PMID- 11421239 TI - Behavioral state-related changes of extracellular serotonin concentration in the pedunculopontine tegmental nucleus: a microdialysis study in freely moving animals. AB - Neurons of the cholinergic mesopontine tegmentum preferentially discharge during REM sleep and are thought to promote this state. It has been hypothesized they are inhibited during wakefulness by serotonergic input. The present study used the microdialysis sampling procedure coupled to microbore HPLC to measure extracellular serotonin levels in the pedunculopontine tegmental nucleus (PPT) in naturally sleeping cats. Extracellular serotonin levels were found to be highest during periods of wakefulness, lower during slow wave sleep, and lowest during periods of REM sleep. During wakefulness serotonin levels (mean A+/-SEM) measured in 10 A microliter samples were 1.14 A+/- 0.13 fmol/sample, whereas during slow wave sleep levels declined significantly to 72% of the wakefulness baseline (0.85 A +/- 0.11 fmol/sample), and dropped further to 45% of the wakefulness baseline in REM samples (0.52 A +/- 0.10 fmol/sample; all p's<0.003). The decrease in PPT serotonin levels during sleep may be an important determinant in the timing of REM sleep cyclicity. The data support the hypothesis that, during slow wave sleep and REM sleep, the declining levels of serotonin release the PPT REM-promoting neurons from serotonergic inhibition, which, in turn, leads to increases in acetylcholine release in terminal areas, facilitating the emergence of REM sleep. PMID- 11421240 TI - Why are doctors so unhappy? It's not all doom and gloom. PMID- 11421241 TI - Bibliography. Current world literature. Emergency and critical care pediatrics and pulmonology. PMID- 11421242 TI - [Cardiogenic shock complicating acute myocardial infarction--etiologies, management and outcome: a report from the SHOCK Trial Registry]. PMID- 11421243 TI - Haplotype analysis involving a novel polymorphism in bovine alpha-lactalbumin 5' flanking region. PMID- 11421244 TI - Plasticity of skeletal myosin in endurance-trained rats (I). A quantitative study. AB - The aim of this study was to quantify modifications in the expression of skeletal myosin light chain (MLC) and myosin heavy chain (MHC) isoforms of five muscles, according to their fiber composition and function, following endurance training in rats. Rodents were assigned randomly to one of two groups: caged sedentary controls (C) or endurance-trained rats (T). In T rats, three out of the four fast and mixed muscles studied exhibited a significant increase in the expression of MLC1s, 1f and 2s and a significant decrease in MLC2f and 3f, the exception being the plantaris muscle. In two out of the four muscles we observed a significant increase in MHCI and IIa, the exception being both gastrocnemii, where the expression of MHCI did not change. In the soleus of T rats, the expression of MLC1s, 2f and 3f decreased significantly, while that of MLC2s increased significantly, compared with those of C rats. The expression of MHCIIa in T rats decreased significantly compared with that of C rats, while the expression of MHCI increased significantly. In all muscles studied, a significant slowing of myosin isoforms was observed after endurance training. PMID- 11421245 TI - Oral fluid therapy of cholera among Bangladesh refugees. 1973. PMID- 11421246 TI - HIV-related diarrhea is multifactorial and fat malabsorption is commonly present, independent of HAART. AB - OBJECTIVE: Highly active antiretroviral therapy (HAART) has significantly decreased the incidence of infectious diarrhea affecting HIV-infected patients. Still, diarrhea remains a common symptom in HIV. We sought to determine the incidence of fat malabsorption as a cause of diarrhea in HIV patients receiving non-HAART (nucleoside analog only) and HAART (protease inhibitor-containing) antiretroviral regimens. METHODS: From June, 1995, to April, 1999, 88 HlV infected patients underwent evaluation for diarrhea, which included endoscopy. We examined the incidence of fat malabsorption with a 24-h stool collection for fecal fat in a cohort of these patients (N = 33). Patients were divided into two groups, those receiving protease inhibitor-containing HAART and those receiving less intensive, nucleoside analog-only, non-HAART regimens. RESULTS: Thirty of 33 patients (90.9%) had fat malabsorption. Twenty of 21 patients not receiving HAART (95.2%) had fat malabsorption with a mean of 34 +/- 38 g of stool fat and a mean stool weight of 797 +/- 454 g. Ten of 12 patients receiving HAART (83.3%) had fat malabsorption with a mean of 46 +/- 86 g of stool fat and a mean stool weight of 800 +/- 647 g. Stool weight correlated with the degree of fat malabsorption (R = 0.77). CONCLUSION: Fat malabsorption represents a commonly undiagnosed entity in HIV-infected patients with diarrhea, whether or not they are receiving HAART therapy. Fecal fat determination should be considered a routine part of the diagnostic workup of HIV-infected patients experiencing diarrhea. PMID- 11421247 TI - Orlistat maintains biliary lipid composition and hepatobiliary function in obese subjects undergoing moderate weight loss. AB - OBJECTIVES: Orlistat, an intestinal lipase inhibitor, has recently been approved by the US Food and Drug Administration for treatment of obesity. The effects of orlistat on hepatobiliary function have not been previously defined. A 4 wk study was performed involving modest weight loss in obese subjects to observe any short term hepatobiliary responses that occur after initiating treatment with orlistat and a hypocaloric diet. METHODS: A total of 23 obese (BMI 30-41 kg/m2) subjects were randomized to a double blind t.i.d. treatment with 120 mg of orlistat or a placebo in conjunction with a hypocaloric diet (1200-1500 kcal/day). The study was designed to achieve similar modest weight loss in both groups in order to be able to directly assess the effects of orlistat. Cholesterol saturation, bile composition, and gallbladder motility were measured. RESULTS: At the end of the treatment period, mean weight loss of 3.8 kg was achieved in the orlistat group (vs 2.3 kg with placebo, p = NS). Total bile acid concentration decreased significantly with placebo (-18.57 +/- 6.99 mmol/L; 95% CI = -32.26 to -4.87), but not with orlistat. Biliary phospholipid concentration decreased significantly with placebo (-4.38 +/- 1.91 mmol/L; 95% CI = -8.13 to -0.64) but not with orlistat. Mean changes from the baseline in cholesterol saturation index and gallbladder motility were similar in both groups. Microscopy of bile failed to reveal cholesterol microcrystals before or after treatment in either group. CONCLUSIONS: Our findings indicate a primary initial effect of weight loss is a reduction in biliary bile acids and phospholipids. Orlistat blocks these adverse changes in biliary lipid composition and maintains hepatobiliary function. We speculate that the risk of formation of gallstones during weight loss may actually be lowered with orlistat. PMID- 11421248 TI - Ionizing radiation, part 2: some internally deposited radionuclides. Views and expert opinions of an IARC Working Group on the Evaluation of Carcinogenic Risks to Humans. Lyon, 14-21 June 2000. PMID- 11421249 TI - Relationships and diseases among drug users and nonusers. AB - Human immunodeficiency virus (HIV) and syphilis are relationship-based diseases that are typically transmitted by the cooperative activities (sex or drug injection) of two persons. A sample of 215 drug users and 52 sociodemographically matched nonusers was collected to examine the behaviors and relationships related to HIV and syphilis transmission. Results showed that, although drug users had more risk opportunities (more sex partners and, of course, more injection partners) than nonusers, actual sex risk behaviors (never using condoms) did not differ appreciably among drug users and nonusers or with opposite-sex partners and same-sex partners. The similarity of sexual risk was supported by the similar levels of syphilis between drug users and nonusers. The unique risk to drug users was drug injection, although drug users were found to engage in fewer risky injection behaviors (sharing of drug injection equipment) than the risky sexual behaviors in which all participants engaged. Although drug users interacted as frequently with partners as nonusers, nonuser relationships were longer lasting and emotionally closer. PMID- 11421250 TI - Children of injection drug users: impact of parental HIV status, AIDS, and depression. AB - We investigated the association between parental factors (including infection with human immunodeficiency virus [HIV], acquired immunodeficiency syndrome [AIDS] diagnosis, parental medical illness, and depression) and children's behavioral and emotional problems among children of injection drug users (IDUs). IDUs were recruited through community outreach. The sample included 73 parents of 73 children, aged 4 to 12 years. Parental depression (odds ratio [OR] = 4.61) and medical illness (OR = 4.70) were found to be significantly associated with internalizing (depressive and anxiety-related symptoms), but not with externalizing (aggressive and disruptive behaviors) symptoms in the children of IDUs. The clinical implications are that children of IDUs are known to be at high risk for psychiatric symptoms and disorders; these data suggest that children of depressed and/or medically ill IDU parents may be at even higher risk of internalizing symptoms (depression and anxiety symptoms) than children of IDUs who do not suffer from these conditions. PMID- 11421251 TI - Muscarinic agonists and antagonists in the treatment of Alzheimer's disease. AB - Alzheimer's disease (AD) is a neurodegenerative disease characterized by cognitive impairment and personality changes. The development of drugs for the treatment of the cognitive deficits of AD has focused on agents which counteract loss in cholinergic activity. Although symptoms of AD have been successfully treated with acetylcholinesterase inhibitors (tacrine, donepezil. rivastigmine, galanthamine), limited success has been achieved with direct M1 agonists, probably due to their lack of selectivity versus other muscarinic receptor subtypes. Muscarinic M2 antagonists have been reported to increase synaptic levels of acetylcholine after oral administration to rats (e.g. BIBN-99, SCH 57790), but their selectivity versus other muscarinic receptor subtypes is modest. Exploration of a series of piperidinylpiperidines has yielded the potent and selective M2 antagonist SCH-217443. This antagonist has excellent bioavailability in rats and dogs and shows activity in a rat model of cognition. PMID- 11421252 TI - Synthesis of 4(1H)-pyridinone derivatives and investigation of analgesic and antiinflammatory activities. AB - This paper describes recent results of a research program aimed at the synthesis and pharmacological evaluation of new 4(1H)-pyridinone derivatives belonging to the 1,3-disubstituted series (4-11). These compounds were structurally planned by applying the molecular hybridization strategy on previously described 1,2 disubstituted-4(1H)-pyridinone derivatives, considered as lead compounds, which present potent analgesic properties (M.D. Aytemir, T. Uzbay, D.D. Erol, Arzneim. Forsch. (Drug Res.) 49 (1999) 250). Their chemical structures have been proved by means of their IR and 1H NMR data and by elemental analysis. The analgesic profile of the title compounds (4-11), evaluated by the model of abdominal constrictions induced by acetic acid, showed that all the 4(1H)-pyridinone derivatives were active, exhibiting an analgesic activity comparable with that of aspirin (acetyl salicylic acid) used as a standard. The antiinflammatory profile of the synthesized compounds, evaluated by the model of carrageenan rat paw edema, showed that all compounds were active and were comparable with indomethacin used as a standard. PMID- 11421253 TI - Glycosyl derivatives of 2-bromosugar of selected non-steroidal anti-inflammatory drugs. Synthesis and QSAR data. AB - On the basis of molecular modeling and calculation of physicochemical SAR data we obtained new derivatives of 2-bromosugars 5a-d and 6 for widely used non steroidal anti-inflammatory drugs (NSAIDs). Compounds 5a-d and 6 were synthesized as stable, crystalline compounds from the reaction of phosphoroates 1-3 with salicylic acid (4a), aspirin (4b), diclofenac (4c) and indomethacin (4d) in an aprotic solvent. The reaction took place in the presence of silver carbonate as the activator of leaving groups with high stereoselectivity and good yields. The structures of the new derivatives of 5a-d and 6 for NSAIDs were established by spectroscopic methods 1H and 13C NMR, and elemental analyses. PMID- 11421254 TI - Synthesis and pharmacological evaluation of aryl/heteroaryl piperazinyl alkyl benzotriazoles as ligands for some serotonin and dopamine receptor subtypes. AB - Thirteen [(aryl/heteroaryl-piperazinyl)alkyl]benzotriazoles were prepared as potential trazodone- and buspirone-like drugs. The synthesized compounds displayed from moderate to good affinity to the serotonin 5-HT1A receptor and only modest or poor affinity to the dopamine D2 receptor, similar to buspirone. The introduction of substituents on the benzotriazole ring did not improve the affinity to the 5-HT1A receptor, compared to the previously described unsubstituted derivatives. In a general pharmacological screening, which concerned only three of these compounds so far (5, 7 and 13), several in vitro and in vivo activities were observed. The guinea pig ileum contractions, induced either electrically or by several agonists, were strongly inhibited; at higher concentrations also the spontaneous tone of the guinea pig trachea was reduced. Compound 13 exhibited good analgesic activity in mice in the formalin-induced algesia and in the writhing test. The same at 30 mg kg(-1) p.o. also displayed antihypertensive activity probably related to calcium channel blockade and adrenergic alpha1 antagonism. In binding assays, 13 showed a IC50 = 580 nM for displacing [3H]prazosin from alpha1 receptor. Finally, compound 5 (and, to a minor extent, compound 13) protected mice against potassium cyanide induced hypoxia. PMID- 11421255 TI - Some reactions with ketene dithioacetals part I: synthesis of antimicrobial pyrazol. AB - Pyrazolo[1,5-a]pyrimidines were synthesized via the reaction of ketene dithioacetals and 5-aminopyrazoles. The antibacterial and antifungal activities of some selected compounds are also reported. PMID- 11421256 TI - Synthesis and analgesic activity of some 1-benzyl-2-substituted-4,5-diphenyl-1H imidazole derivatives. AB - In this study, derivatives of 1-benzyl-2-substituted-4,5-diphenyl-1H-imidazole were synthesized and their analgesic activity assayed in two tests. 1,2,4,5 Tetrasubstituted imidazole compounds were obtained by the treatment of purified imidazole compounds with benzyl chloride in the presence of sodium hydride. The structure elucidation of the compounds was performed by IR, 1H-NMR and mass spectroscopic data and elemental analysis results. Generally the prepared compound exhibited only moderate analgesic activity in mice at the dose of 100 mg/kg i.p.; however, a few of them exhibited good activity, almost equivalent to that of morphine at 1 mg/kg i.p. was observed. At the above dosage, no toxicity was observed for all compounds. PMID- 11421257 TI - Use of 7-fluoro-4-nitrobenzo-2-oxo-1,3-diazole (NBD-F) for the determination of ramipril in tablets and spiked human plasma. AB - Ramipril, as a secondary amine compound, reacts with 7-fluoro-4-nitrobenzo-2-oxo 1,3-diazole (NBD-F) producing the corresponding fluorescent NBD-ramipril. According to this fact, spectrophotometric and fluorimetric methods for the determination of ramipril were developed. The effect of these parameters on the reaction product were carefully studied to optimize reaction conditions. The relationship between the absorbance at 465 nm and the concentration was found to be linear over the range 1-10 microg/ml. Moreover, the fluorescence intensity was also found to be directly proportional at the concentration over the range of 20 100 ng/ml at 530 nm after excitation at 465 nm. The proposed procedure was successfully applied to the determination of ramipril in both tablet dosage form and in plasma. Spectrophotometric determination of ramipril tablets yielded a percentage recovery of 98.66+/-0.38, while the percentage recovery of spectrofluorimetric determination of ramipril in spiked human plasma was 99.08+/ 1.11%. The results obtained are in good agreement with those obtained by the reference method. No interference could be observed from the co-administered drug (hydrochlorothiazines). PMID- 11421258 TI - Cytotoxic cyclolignans related to podophyllotoxin. AB - The cyclolignan family of natural products includes compounds with important antineoplastic and antiviral properties such as podophyllotoxin and two of their semisynthetic derivatives, etoposide and teniposide. The latter are included in a wide variety of cancer chemotherapy protocols. Due to these biological activities, cyclolignans have been the objective of numerous studies focused to prepare better and safer anticancer drugs. Several cyclolignans related to podophyllotoxin have been prepared and evaluated for their cytotoxic activities on four neoplastic cell lines (P-388, A-549, HT-29 and MEL-28); some of them have antiviral and immunosuppressive activities. PMID- 11421259 TI - Aromatic sialic acid analogues as potential inhibitors of influenza virus neuraminidase. AB - The influenza virus neuraminidase (NA) is an enzyme essential for viral infection and offers a potential target for antiviral drug development. We aimed our research at the synthesis of non-carbohydrate molecules able to inhibit NA as transition-state analogues. Aromatic sialic acid analogues (compound 5 and compound 10) were synthesised in good yields starting from commercially available benzoic acids using a suitable synthetic strategy. PMID- 11421260 TI - Synthesis and antiplatelet activity of some 2,7-di(N-cycloamino)-3-phenyl-1,8 naphthyridine derivatives. AB - Several 2,7-di(N-cycloamino)-3-phenyl-1,8-naphthyridine derivatives were synthesized and tested for their ability to inhibit human platelet aggregation in vitro induced by arachidonic acid, collagen and ADP. Only five compounds showed any appreciable activity, and the results of all the active derivatives were similar to those of papaverine in the test with arachidonic acid and collagen. Moreover, the most active compounds were investigated in the test with ADP and again showed a significant activity. The tested compounds that possessed the best activity were also shown to increase the c-AMP level significantly without involving the adenylyl cyclase system. PMID- 11421261 TI - Synthesis and biological activity of glycosyl conjugates of N-(4 hydroxyphenyl)retinamide. AB - Three glycosyl (glucosyl, galactosyl, mannosyl) conjugates of 4 hydroxyphenylretinamide have been synthesized and tested on a broad variety of tumor cells. All three compounds are active on promyelocytic leukemia cell lines HL60 but less than the parent compound 4-HPR. Among them, the mannosyl analog stands out by its very low toxicity. PMID- 11421262 TI - Synthesis and antimicrobial properties of N-substituted derivatives of (E)-4 azachalcones. AB - Syntheses of 11 new N-bromoalkyl substituted bromides of (E)-4-azachalcone and N o-(m- and p-) halobenzyl substituted halides of (E)-3'-hydroxy-4-azachalcone of antimicrobial activity are reported. Compounds 3d, 3e, 6b, 6c, and 6e reveal good antimicrobial activity against Staphylococcus aureus and Enterococcus faecalis as well as moderate activity against Escherichia coli and Klebsiella pneumoniae. PMID- 11421263 TI - Synthesis and biological evaluation of pseudostellarin B. AB - A new potent bioactive cyclic peptide pseudostellarin B has been synthesised. The structure was elucidated by elemental analyses, IR, 1H NMR, 13C NMR and FAB mass spectral data. The synthesised compound was also screened for its antibacterial, antifungal, antiinflammatory and anthelmintic activities. PMID- 11421264 TI - Anti-inflammatory effect of Pimenta racemosa var. ozua and isolation of the triterpene lupeol. AB - Pimenta racemosa var. ozua (Myrtaceae) is a tropical plant, used in different inflammatory processes by the folk medicine of the Caribbean region. From the methanol extract of the leaves a terpenic compound identified as lupeol has been isolated for the first time in this species. The anti-inflammatory activity of the extract has been evaluated against two experimental models of acute inflammation: paw edema in rats, using carrageenan or dextran as phlogogen agents, and ear edema in mice, inducing the inflammation with 12-o tetradecanoylphorbol acetate (TPA). Myeloperoxidase activity (MPO) was also assayed as an indicator of leukocytary migration in the inflamed ears. In the carrageenan test, the methanol extract (125 and 250 mg kg(-1) p.o.) had a dose dependent and significant effect at different time intervals. On the contrary, when the dextran was injected in paw, the extract did not reduce the inflammation provoked. This behavior was similar to indomethacine (25 mg kg(-1)) used as a standard drug. In the TPA-induced ear edema, the methanol extract (0.5, 1 and 3 mg ear(-1)) significantly reduced the inflammation. In the MPO assay a significant inhibition of the enzyme was observed in the inflamed tissue in all the samples assayed. These results show that the methanol extract from the leaves of Pimenta racemosa var. ozua, is effective against acute inflammation processes, by oral route and when topically applied. The anti-inflammatory behavior of the extract was similar to that exhibited by the selective cyclo-oxygenase inhibitor, indomethacin. On the other hand, the reduction of MPO activity shows that the action mechanism is clearly related with the neutrophil migration. PMID- 11421265 TI - Muslim families: Donating organs and asking for post mortems. PMID- 11421266 TI - Pediatric ethics in the Holy Quran. PMID- 11421267 TI - Medicines for children and gentamicin toxicity. PMID- 11421268 TI - The rise of a protein family: ATP-binding cassette systems. PMID- 11421269 TI - ABC transporters: physiology, structure and mechanism--an overview. AB - ABC transporters form one of the largest of all protein families with a diversity of physiological functions. In Escherichia coli almost 5% of the genome is occupied by genes encoding components of these transporters, and there are examples in all species from microbes to man. In this overview, the importance of studies on bacteria in elucidating many basic principles pertaining to ABC transporters is emphasised. The family is described and a general overview of the structure and function of these transporters is presented. PMID- 11421270 TI - The ABC of ABCS: a phylogenetic and functional classification of ABC systems in living organisms. AB - ATP binding cassette (ABC) systems constitute one of the most abundant superfamilies of proteins. They are involved not only in the transport of a wide variety of substances, but also in many cellular processes and in their regulation. In this paper, we made a comparative analysis of the properties of ABC systems and we provide a phylogenetic and functional classification. This analysis will be helpful to accurately annotate ABC systems discovered during the sequencing of the genome of living organisms and to identify the partners of the ABC ATPases. PMID- 11421271 TI - A new family of high-affinity ABC manganese and zinc permeases. AB - Phylogenetic analysis of 47 extracellular putative metal binding receptors (MBRs) belonging to the newly defined cluster suggests the existence of two subclusters. The question of substrate specificity of the corresponding ATP binding cassette (ABC) permeases is discussed, based on data collected from 19 of them concerning their regulation, metal requirement of permease mutants, metal uptake and metal binding. The proposal that the two subclusters correspond to paralogous metal permeases dedicated primarily to manganese and to zinc transport is made. The question of a direct role of MBRs as adhesins of Gram-positive bacteria is then discussed and the importance of metal permeases for cellular processes and host bacteria interactions is reviewed. PMID- 11421272 TI - Peptides and ATP binding cassette peptide transporters. AB - In this review our knowledge of ATP binding cassette (ABC) transporters specific for peptides is discussed. Besides serving a role in nutrition of the cell, the systems participate in various signaling processes that allow (micro)organisms to monitor the local environment. In bacteria, these include regulation of gene expression, competence development, sporulation, DNA transfer by conjugation, chemotaxis, and virulence development, and the role of ABC transporters in each of these processes is discussed. Particular attention is paid to the specificity determinants of peptide receptors and transporters in relation to their structure and to the mechanisms of peptide binding. PMID- 11421273 TI - Bacterial ABC transporters of amino acids. AB - There are two subfamilies of ABC uptake systems for amino acids in bacteria, the polar amino acid transport family and the hydrophobic amino acid transport family. We consider the general properties of these families and we examine the specific transporters. Focusing on some of the best-studied ATP binding cassette transporters we also examine the mechanism of amino acid uptake, paying particular attention to the question of bidirectionality of solute movement. PMID- 11421274 TI - Polyamine uptake systems in Escherichia coli. AB - The polyamine content of cells is regulated by biosynthesis, degradation, and transport. In Escherichia coli, the genes for three different polyamine transport systems have been cloned and characterized. Two uptake systems (putrescine specific and spermidine-preferential) are ABC transporters, each consisting of a periplasmic substrate binding protein, two transmembrane proteins, and a membrane associated ATPase. The third transport system, catalyzed by PotE, mediates both uptake and excretion of putrescine. In this article, the properties of the first two polyamine uptake systems are reviewed in detail. PMID- 11421275 TI - Bacterial transporters for sulfate and organosulfur compounds. AB - Microorganisms require sulfur for growth, and obtain it either from inorganic sulfate or from organosulfur compounds such as sulfonates, sulfate esters, or sulfur-containing amino acids. Transport of sulfate into the cell is catalyzed either by ATP binding cassette (ABC)-type transporters (SulT family) or by major facilitator superfamily-type transporters (SulP family). By contrast, the sulfonate and sulfate ester transporters identified to date are all ABC-type systems, whose synthesis is tightly regulated by the sulfur supply to the cell, mediated by the CysB protein and other transcriptional regulators of the LysR family. PMID- 11421276 TI - ABC transporter-mediated uptake of iron, siderophores, heme and vitamin B12. AB - Microbes have developed a number of different strategies to utilize iron, which is a vital element for most organisms but not always readily available from the environment. Based on experimental studies and sequence analysis data, this article gives a short overview of ABC transporters related to iron uptake: components of three distinct families mediate the translocation of iron, siderophores, heme and vitamin B12 across the cytoplasmic membrane of bacteria. PMID- 11421277 TI - ABC transporters catalyzing carbohydrate uptake. AB - ATP binding cassette (ABC) transporters mediating the uptake of carbohydrates comprise two subfamilies (CUT1, CUT2) that differ with respect to the chemical nature of their substrates, subunit composition, and conserved sequence motifs. In this article, current knowledge of members of each family is summarized with special emphasis on the well-characterized transport systems for maltose/maltodextrin and ribose, respectively, of enterobacteria. PMID- 11421278 TI - Molybdate transport. AB - In both bacteria and archaea, molybdate is transported by an ABC-type transporter comprising three proteins, ModA (periplasmic binding protein), ModB (membrane protein) and ModC, the ATPase. The modABC operon expression is controlled by ModE Mo. In the absence of the high-affinity molybdate transporter, molybdate is also transported by another ABC transporter which transports sulfate/thiosulfate as well as by a nonspecific anion transporter. Comparative analysis of the molybdate transport proteins in various bacteria and archaea is the focus of this review. PMID- 11421279 TI - ABC transporters associated with cytochrome c biogenesis. AB - It is generally agreed that cytochrome c biogenesis requires that the apocytochrome and heme be transported separately to their site of function and assembly. In bacteria, this is outside the cytoplasmic membrane, whereby the apocytochromes c use sec-dependent signals for their translocation. Two different hypotheses have recently emerged as to how heme is exported: one involves an helABCD-encoded ATP binding cassette (ABC) transporter complex and the second does not. The second hypothesis concludes that an (HelAB)2 heterodimeric ABC transporter does not transport heme but some other substrate for cytochrome c biogenesis. The evidence supporting each of these two hypotheses and the role of this ABC transporter is discussed. PMID- 11421280 TI - Mitochondrial ABC transporters. AB - In contrast to bacteria, mitochondria contain only a few ATP binding cassette (ABC) transporters in their inner membrane. The known mitochondrial ABC proteins fall into two major classes that, in the yeast Saccharomyces cerevisiae, are represented by the half-transporter Atm1p and the two closely homologous proteins Mdl1p and Mdl2p. In humans two Atm1p orthologues (ABC7 and MTABC3) and two proteins homologous to Mdll/2p have been localized to mitochondria. The Atm1p like proteins perform an important function in mitochondrial iron homeostasis and in the maturation of Fe/S proteins in the cytosol. Mutations in ABC7 are causative of hereditary X-linked sideroblastic anemia and cerebellar ataxia (XLSA/A). MTABC3 may be a candidate gene for the lethal neonatal syndrome. The function of the mitochondrial Mdl1/2p-like proteins is not clear at present with the notable exception of murine ABC-me that may transport intermediates of heme biosynthesis from the matrix to the cytosol in erythroid tissues. PMID- 11421281 TI - The role of ABC transporters in antibiotic-producing organisms: drug secretion and resistance mechanisms. AB - Knowledge about biosynthetic gene clusters from antibiotic-producing actinomycetes is continuously increasing and the presence of an ABC transporter system is a fairly general phenomenon in most of these clusters. These transporters are involved in the secretion of the antibiotic through the cell membrane and also contribute to self resistance to the produced antibiotic. PMID- 11421282 TI - ABC transporters of staphylococci. AB - Some members of the genus Staphylococcus are important human pathogens, such as Staphylococcus aureus and Staphylococcus epidermidis. ATP binding cassette (ABC) transporters of pathogenic staphylococci are often responsible for antibiotic resistance and for the uptake of essential solutes and are therefore being considered as targets for novel antistaphylococcal drugs. Other ABC transporters are involved in the biosynthesis of a special class of peptide antibiotics, the lantibiotics, and in self-protection of the producer against them. PMID- 11421283 TI - ABC transporters and the export of capsular polysaccharides from gram-negative bacteria. AB - In this review, we discuss the kps cluster of Escherichia coli as the paradigm for the ABC capsular polysaccharide exporter (CPSE) family. Components of the cluster form a multimeric protein complex consisting of both biosynthetic and export machinery. We compare the Kps exporter with capsule export systems from other members of the CPSE family. PMID- 11421284 TI - Multidrug transport by ATP binding cassette transporters: a proposed two-cylinder engine mechanism. AB - The elevated expression of ATP binding cassette (ABC) multidrug transporters in multidrug-resistant cells interferes with the drug-based control of cancers and infectious pathogenic microorganisms. Multidrug transporters interact directly with the drug substrates. This review summarizes current insights into the mechanism(s) by which ATP hydrolysis is coupled to drug transport in bacterial LmrA and its human homolog P-glycoprotein. In addition, the relevance of these insights for other ABC transporters will be discussed. PMID- 11421285 TI - Fungal ABC proteins: pleiotropic drug resistance, stress response and cellular detoxification. AB - A number of prominent genetic diseases are caused by mutations in genes encoding ATP-binding cassette (ABC) proteins (Ambudkar, Gottesmann, 1998). Moreover, several mammalian ABC proteins such as P-glycoprotein (P-gp) (Gottesman et al., 1995) and multidrug-resistance-associated proteins (MRPs) (Cole, Deeley, 1998) have been implicated in multidrug resistance (MDR) phenotypes of tumor cells highly resistant to many different anticancer drugs. The characteristics of MDR phenomena include the initial resistance to a single anticancer drug, followed by the development of cross-resistance to many structurally and functionally unrelated drugs. Similar mechanisms of MDR exist in pathogenic fungi, including Candida and Aspergillus (Vanden Bossche et al., 1998), and also in parasites such as Plasmodium and Leishmania (Ambudkar, Gottesmann, 1998), as well as in many bacterial pathogens (Nikaido, 1998). To dissect the mechanisms of MDR development and to elucidate the physiological functions of ABC proteins, many efforts have been made during the past decade. Importantly, yeast orthologues of mammalian disease genes made this unicellular eukaryote an invaluable model system for studies on the molecular mechanisms of ABC proteins, in order to better understand and perhaps improve treatment of ABC gene-related disease. In this review, we provide an overview of ABC proteins and pleiotropic drug resistance in the budding yeast Saccharomyces cerevisiae and the fission yeast Schizosaccharomyces pombe. Furthermore, we discuss the role of ABC proteins in clinical drug resistance development of certain fungal pathogens. PMID- 11421286 TI - Translational regulation by ABC systems. AB - Elongation factor 3 is a cytosolic protein required by the fungal ribosomes for in vitro protein synthesis and for in vivo growth. EF-3 stimulates binding of EF 1:GTP:aa-tRNA ternary complex to the ribosomal A site by facilitated release of the deacylated tRNA from the E site. The reaction requires ATP hydrolysis. EF-3 contains two ATP binding sequence (NBS) motifs. NBSI is sufficient for the intrinsic ATPase activity. NBSII is essential for the ribosome-stimulated functions. PMID- 11421287 TI - Role of ATP hydrolysis by UvrA and UvrB during nucleotide excision repair. AB - Nucleotide excision repair in eubacteria is a process that repairs DNA damages by the removal of a 12-13-mer oligonucleotide containing the lesion. Recognition and cleavage of the damaged DNA is a multistep ATP-dependent reaction that requires the UvrA, UvrB and UvrC proteins. Both UvrA and UvrB are ATPases, with UvrA having two ATP binding sites which have the characteristic signature of the family of ABC proteins and UvrB having one ATP binding site that is structurally related to that of helicases. PMID- 11421288 TI - Enhanced low shear stress induced platelet aggregation by Shiga-like toxin 1 purified from Escherichia coli O157. AB - The effect of Shiga-like toxin 1 (Stx1) produced by Escherichia coli O157 on platelets was studied with an argon laser light-assisted shear-induced platelet aggregometer and with binding assays. Stx1 markedly enhanced the platelet aggregation under low shear stress but did not affect it under high shear stress. Minimal concentration of Stx1 required for the enhancement was 0.25 ng/ml, and almost maximal enhancement was observed at a final concentration of > or =2.5 ng/ml. This enhanced platelet aggregation disappeared after leukocyte depletion from normal platelet-rich plasma with a specific filter. In contrast, a standard platelet aggregometer was unable to detect this enhanced platelet aggregation in either the presence or the absence of ADP. 125I-labeled purified Stx1 did not specifically bind to normal washed platelets depleted of leukocytes, and thin layer chromatographic analysis of glycolipids extracted from normal platelet lysates also confirmed that leukocyte-depleted normal platelets lack Stx1 specific receptor globotriaosylceramide (Gb3). Supernatant from the monocyte suspension stimulated with Stx1 exhibited the enhanced low shear stress induced platelet aggregation, but that from the polymorphonuclear cell suspension did not. Several cytokines produced from monocytes reproduced this event in vitro. Further, plasmas from six out of seven patients with hemolytic uremic syndrome (HUS) had activity similar to the purified Stx1. This activity was almost totally impaired after treatment of HUS plasmas with Gb3 in accord with reduction of plasma Stx1 levels. Taken together, these results indicate that platelets lack Gb3, and Stx1 appears to modulate platelet aggregation in an indirect fashion, presumably by the release of cytokines or chemical compounds from the target tissues. PMID- 11421289 TI - Autoimmune thrombocytopenia in Waldenstrom's macroglobulinemia. AB - Autoimmune phenomena are well-recognised complications of Waldenstrom's macroglobulinemia (WM) and IgM monoclonal gammopathy. Peripheral neuropathy and cold agglutinin hemolytic anemia are the most common reported and occur in 5-10% of patients. Autoimmune thrombocytopenia has been rarely reported in WM and its incidence is not known. In this study we report the case of a 67-year-old man who presented with autoimmune thrombocytopenia who was subsequently found to have WM. Laboratory investigation demonstrated that platelet-associated IgM (PAIgM) but not PAIgG was clearly elevated compared to normal controls. In addition the patient's serum reacted strongly with a panel of donor platelets analysed with an indirect platelet immunofluorescence assay utilising an anti-IgM secondary antibody. Glycoprotein specificity could not however be demonstrated by ELISA techniques for platelet glycoproteins IIbIIIa, IaIIa, IbIXa, and IV. We also reviewed the case records of 104 additional cases of WM diagnosed at our institution between 5/93 and 5/99. Three further cases with clinically significant autoimmune thrombocytopenia were identified. The overall incidence of autoimmune thrombocytopenia (4/105, 3.8%) in this cohort of patients was similar to the incidence of peripheral neuropathy (7/105, 6.7%) and cold agglutinins (3/105, 2.9%). PMID- 11421290 TI - Monocytic leukemia cutis diagnosed simultaneously with refractory anemia with monocytosis: a case report. AB - A case of leukemia cutis (LC) of monocytic lineage in a patient with myelodysplastic syndrome (MDS) is presented. Cutaneous infiltrates were recognized concurrent with diagnosis of refractory anemia (RA) with monocytosis. Skin infiltrates subsequently spontaneously regressed although MDS progressed with increasing monocytosis, anemia, and thrombocytopenia. Death occurred 6 months after diagnosis with evolution of acute monoblastic leukemia complicated by sepsis. This case supports previous observations of poor prognosis associated with leukemia cutis. LC associated with MDS is reviewed including the role of monocytes. PMID- 11421291 TI - Multiple myeloma and human immunodeficiency virus-1 (HIV-1) infection. AB - Multiple myeloma (MM) in three human immunodeficiency virus (HIV)-infected patients is reported. HIV infection predisposes to the development of high-grade B-cell lymphomas, but few cases of plasma cell tumours in association with HIV have been reported. The coincidence of HIV infection and neoplasia highlights the distinct roles of immunodeficiency and infection with herpesviridae, including HIV itself, in the pathogenesis of HIV-related tumours. In addition, a number of cytokines (e.g., interleukin-6 [IL-6]) and angiogenic factors (e.g., vascular endothelial growth factor [VEGF] and basic fibroblastic growth factor [bFGF]) may play a role in the initiation, maintenance, and progression of multiple myeloma (MM). Infection was the first clinical consideration to the cause of the illness in two of our HIV-seropositive patients. The diagnosis of MM may be difficult in patients with advanced HIV infection as they often have renal failure, bone marrow plasmacytosis, repeated infections, and polyclonal hypergammaglobulinaemia, due to HIV infection itself, opportunistic pathogens, and/or medication. PMID- 11421292 TI - Case report of sideroblastic anemia caused by ingestion of coins. AB - This is a report of a 54-year-old schizophrenic patient with a 15-year history of ingesting metal objects (pica). He presented with severe anemia (hemoglobin of 3 g/dl and hematocrit of 8.3%) and leukopenia (white blood count of 1,300/mm3). Work-up revealed copper deficiency (copper level of <0.05 microg/ml) and elevated zinc levels (280 microg/ml). The zinc toxicity was produced by the zinc content in the coins ingested by the patient over a period of many years. He was initially treated with -acetylcysteine and sodium bicarbonate followed by intravenous copper sulfate. He was also placed on Adolph's meat tenderizer and pancreatin thrice a day orally to loosen the massive amount of metallic objects including coins in his bowel and allow them to pass out in his feces. He was also continued on oral copper sulfate. His copper levels began to rise and reached a maximum of 0.72 microg/ml, and his zinc level fell to 153 microg/ml. However, as he refused surgery to remove the metal objects from his bowel and continued to ingest more coins, there was continued absorption of zinc, which later overcame the efforts to reduce the zinc level and increase copper levels in his blood. He finally succumbed to sepsis and multiorgan failure. Autopsy revealed a coin mass in the stomach weighing 1,870 grams in addition to a sigmoid volvulus caused by another coin bezoar in the colon. PMID- 11421293 TI - Platelet factor V New York: a defect in factor V distinct from that in factor V Quebec resulting in impaired prothrombinase generation. AB - Studies were performed on a patient with a longstanding bleeding disorder whose major defects were impaired platelet prothrombinase activity in the absence of added factor Va, and a platelet factor V value that was either decreased or at the lower limit of normal when assayed on multiple occasions. In contrast, plasma factor V values were consistently normal. Unlike Scott Syndrome, in which platelet prothrombinase activity is decreased in both the presence and absence of added factor V, her platelets appeared to utilize added factor Va normally in supporting the generation of prothrombinase activity. These findings suggest an intrinsic defect in platelet factor V as the basis of her platelet prothrombinase defect. This defect appears to be different than that described in the Quebec platelet disorder (factor V Quebec). Immunoblot analyses of washed platelet lysates demonstrated a pattern of variably sized factor V molecules that was entirely similar to that observed in normal platelets, and both the heavy and light chains of her factor V after thrombin cleavage were of the same size as that observed in normal platelets. In addition, her platelet multimerin was normal and immunoblot analysis excluded the type of generalized granular protein defect and pathological proteolysis that has been suggested to explain the factor V defect in the Quebec platelet disorder. The findings in this patient thus suggest a new type of platelet factor V defect as the basis for the impaired capacity of her activated platelets to support prothrombinase generation. The findings further support an important role for platelet factor V in hemostasis. PMID- 11421294 TI - Treatment of Kasabach-Merritt syndrome by embolisation of a giant liver hemangioma. AB - We report the case of a 14-month-old child with Kasabach-Merritt Syndrome, due to a giant liver hemangioma. The therapeutic approach consisted of peripheral transcatheter embolisation of the right hepatic artery with Ivalon microspheres without the addition of thrombogenic material. This procedure brought to a sensible permanent reduction of the size of the liver hemangioma with normalisation of the previous altered coagulation parameters after 6 years of follow-up. PMID- 11421295 TI - Successful treatment of paraneoplastic pemphigus in follicular NHL with rituximab: report of a case and review of treatment for paraneoplastic pemphigus in NHL and CLL. AB - Paraneoplastic pemphigus is a severe mucocutaneous disease associated in most cases with B-cell lymphoproliferative disorders. Independent of the course of the underlying malignancy, this autoimmune skin disease is resistant to any treatment in most cases and may lead to death by infectious complications. We observed a patient with recovery of paraneoplastic pemphigus associated with follicular non Hodgkin's-lymphoma after therapy with rituximab. In addition, we reviewed the literature and analyzed the few cases with comparable outcome with other treatment modalities. PMID- 11421296 TI - Familial nasal NK/T-cell lymphoma and pesticide use. AB - Familial occurrence of nasal NK/T-cell lymphoma (NNKTCL) in pesticide users is presented. The proband (71 years old, male) and son (39 years old) were both diagnosed with NNKTCL within interval of 26 months. Laboratory data showed slight anemia, with no abnormal cells in peripheral blood. They and their wives were farmers and used large amounts of pesticides (fungicides and insecticides) in the hothouse. NNKTCL did not develop in the wives. Proband's father was diagnosed with malignant lymphoma of the neck and died of the disease. Genetic analyses of the peripheral blood leukocytes and tumor tissues did not show p53 and k-ras gene mutations and microsatellite instability. Metaphase cells from peripheral blood leukocytes bore specific marker chromosomes (father, 44XY,-14,-17,-18,-22,+2mar; son, 46XY,-17,+1mar). Environmental exposures to pesticides in conjunction with familial or genetic factors might increase the risk for NNKTCL. PMID- 11421297 TI - Treatment of HIV-associated multicentric Castleman's disease with oral etoposide. AB - Multicentric Castleman's disease (MCD) is a lymphoproliferative disorder that can be defined based upon both clinical and pathological characteristics. The clinical features of this frequently fatal disease include fever, generalized lymphadenopathy, fatigue, splenomegaly, hepatomegaly, and pancytopenia. Recently, severe forms of this disease have been diagnosed in HIV positive patients. Human herpesvirus type 8 (HHV-8) DNA sequences have been detected in peripheral blood mononuclear cells (PBMCs) of patients with Kaposi's sarcoma and MCD, regardless of HIV infection status. Treatment and outcomes in HIV associated MCD are generally unfavorable. We recently treated two HIV-positive patients diagnosed with aggressive MCD with daily oral etoposide (50 mg). The first patient had relapsed on several occasions despite previous therapy with doxil, paclitaxel, and oral ganciclovir. The second patient was treatment naive. Both patients had HHV-8 detectable by polymerase chain reaction in PBMCs, widespread tumor, and B type symptoms when therapy was initiated. In both cases remissions (documented by computerized tomography) have been durable, 1.5 and 6 months, respectively, with minimal side effects. Oral etoposide may be a safe, tolerable, and active agent in MCD. PMID- 11421298 TI - Intravenous gammglobulin-associated acute renal failure. AB - Intravenous gammaglobulins are used for the treatment of various auto-immune hematological disorders. Renal failure is a relatively rare, but an increasingly recognized side effect of gammaglobulin therapy. Although the renal failure is usually reversible, renal replacement therapy is required occasionally. A high index of suspicion, early recognition and appropriate intervention can prevent this complication. We herewith describe two patients with an immune hematological disorder, who developed acute renal failure after treatment with intravenous gammaglobulins. A brief review of the possible risk factors, pathophysiology and management of this complication is provided. PMID- 11421299 TI - Mild hyperhomocysteinemia and penile venous thrombosis. PMID- 11421300 TI - Successful treatment of acquired pure red cell aplasia and acquired amegakaryocytic thrombocytopenia with anti-thymocyte globulin. PMID- 11421301 TI - Quality of life improvement for patients with sickle cell disease. PMID- 11421302 TI - Intense immunosuppressive therapy followed by autologous peripheral blood selected progenitor cell reinfusion for severe autoimmune disease. AB - Autologous stem cell transplantation (HSCT) has been shown to be effective in curing a large spectrum of autoimmune disorders. Case reports are being collected in the EBMT/EULAR Autoimmune Disease Stem Cell Project registry, which reports transplant-related mortality (TRM) of 6%. In order to reduce TRM and preserve the anti-autoimmune effect we evaluated a more immunoablative as opposed to myeloablative conditioning regimen for the autotransplant of severe immunomediated diseases. We enrolled patients affected by systemic lupus erythematosus (SLE: 3 patients), by autoimmune thrombocytopenic purpura (AITP: one patient), by thrombotic thrombocytopenic purpura (TTP: one patient), by pure red cell aplasia (PRCA: one patient), and by a severe cryoglobulinemia (one patient). All patients were mobilized with cyclophosphamide (Cy) 4 g/m2 + G-csf. Conditioning regimen consisted of Cy 50 mg/kg/day (days -6 and -5); anti-T globulin (ATG) 10 mg/kg/day and 6-methylprednisolone (PDN) 1 g/day (days -4, -3, and -2). Immunomagnetically selected CD34+ cells were re-infused on day 0. In three patients neutrophil count fell below 0.5 x 10(9)/l, while a PLT count below 20 x 10(9)/l was registered in two patients. Extrahematological toxicity was very low. Four patients (2 SLE, 1 TTP, 1 cryoglobulinemia) are in complete corticosteroid-free remission with a median follow up of 335 days. The third SLE patient improved considerably; however, he still needs low-dose corticosteroid maintenance. The AITP and PRCA patients achieved a CR but soon relapsed; nevertheless, the procedure restored a steroid-sensitive status. The use of this immunoablative conditioning regimen in auto-HSCT transplant was shown to be effective in controlling disease progression and could be a valuable strategy in reducing TRM. PMID- 11421303 TI - Successful treatment of refractory anemia by high-dose methylprednisolone associated with an increment in CD68-positive cells in bone marrow. AB - Refractory anemia has a relatively low incidence of the subsequent development of acute leukemia and a relatively long survival among the myelodysplastic syndromes (MDS). We observed hematological improvement due to high-dose methylprednisolone in 9 of 18 patients with refractory anemia. The patients' age range was from 28 to 78 years old (mean age: 54), including 14 male and 4 females. A complete response was obtained in 5 patients, minimal response in 4 patients, and no response in 9 patients. Laboratory data of peripheral blood counts and differential counts of bone marrow aspirates were not different, except that fewer chromosomal abnormalities (P = 0.086) were observed in the responder group. Side effects were seen in two patients but were controllable. Overall survival was significantly longer in the responder group (Log-rank P = 0.040, Wilcoxon P = 0.045). The overall survival of responders did not reach the median and 85% of the patients were alive after 180 months, while the median overall survival of the nonresponders was 61.8 months. Disease progression was more frequently seen in the non-responder group (P = 0.045). Furthermore, we investigated retrospectively immunohistochemical bone marrow staining, and a significantly higher percentage of CD68-positive (22.6% +/- 7.1%) and CD45RA-positive cells was observed in the responder group compared to the non-responder group (6.5% +/- 1.3%). Our present results indicate that high-dose methylprednisolone is valuable as a primary treatment before other immunosuppressive treatments, because of its ease of use. High efficacy with high-dose methylprednisolone is expected, especially in patients in which increments in CD68-positive cells in bone marrow are observed. PMID- 11421304 TI - Serum levels of fluconazole in patients after cytotoxic chemotherapy for hematological malignancy. AB - We performed a prospective evaluation of pharmacokinetics of fluconazole administered for prophylactic purposes to 19 patients after cytotoxic chemotherapy for hematological malignancies. On days 7 and 15, we obtained 5 ml of blood from each patient. If fluconazole was administered orally, blood samples were drawn 2, 8, and 24 hr after ingestion of the drug. If it was administered intravenously, blood samples were drawn 1, 8, and 24 hr post-injection. Serum fluconazole levels were analyzed by HPLC with ultraviolet light detection. In patients receiving 200 or 400 mg of fluconazole per day, maximal serum levels were 7.9 and 15.6 mg/l and minimum levels were 5.0 and 10.3 mg/l, respectively. There was no significant difference in serum fluconazole levels comparing the levels after oral and intravenous administration, and pharmacokinetic parameters of fluconazole were comparable at each time point within one dose level. However, considerable variation in serum fluconazole levels was noted in this study, as the maximal serum levels ranged from 4.0 to 13.3 mg/l and from 8.7 to 26.9 mg/l in patients receiving 200 and 400 mg of fluconazole orally, respectively. These variations may be associated with prophylactic failures for patients with insufficient fluconazole concentrations. Multiple regression analysis showed significant correlation between serum fluconazole levels and some variables including dose of fluconazole, age, serum aspartate aminotransferase levels and blood urea nitrogen levels. These variations may be associated with disturbance of body water balance, such as massive hemorrhage and dehydration. PMID- 11421305 TI - Response to measles, mumps, and rubella revaccination among HIV-positive and HIV negative children and adolescents with hemophilia. Hemophilia Growth and Development Study. AB - The effect of human immunodeficiency virus (HIV) infection on response to measles, mumps, and rubella revaccination in children and adolescents with hemophilia was evaluated. Antibody levels of measles, mumps, and rubella were assayed at baseline and two annual examinations in 207 HIV-positive and 126 HIV negative hemophiliacs participating in the Hemophilia Growth and Development Study (HGDS). Response to revaccination was analyzed for participants whose antibody levels were below the cut-off at the start of a year-long observation period. Among HIV-positive participants, antibody levels were below cut-off in 52 subjects for measles, in 71 for mumps, and in 96 for rubella. Among HIV-negative participants, antibody levels were low in 23 subjects for measles, in 23 for mumps, and in 31 for rubella. For measles and mumps antigens, revaccination was associated with a significant increase in redraw antibody levels for HIV-negative participants. Although there was an increase in the mean measles titers for revaccinated HIV-positive participants, it was not significant. Revaccination was associated with an increase in rubella antibodies in HIV-positive and HIV negative participants. Revaccination with measles and mumps was associated with an increase in antibody levels in HIV-negative participants but not in HIV positive participants. Both HIV-positive and HIV-negative participants responded to rubella revaccination with an increase in antibody levels. PMID- 11421306 TI - Agamma-haplotypes: a new group of genetic markers for thalassemic mutations inside the 5' regulatory region of the human Agamma-globin gene. AB - This study illustrates the relationship between a group of nucleotide variations within the 5' regulatory region of the Agamma-globin gene [Agamma-588 A-->G, Agamma-499 T-->A and the 4-bp deletion (Agamma-225 to -222 AGCA)] and the spectrum of delta- and beta-thalassemia mutations in the Hellenic population. These sequence variations, screened by means of denaturing gradient gel electrophoresis, form four separate frameworks (Agamma-haplotypes), each one of which was found to be linked in cis with certain delta- and beta-thalassemia mutations found in the Hellenic population. In addition, two novel base substitutions inside the 5' regulatory region of the Agamma-globin gene (Agamma 521 C-->A and Ay-500 C-->T) were identified during this study, which together with Agamma-haplotypes seem to be silent polymorphisms during adult life, as indicated by transient expression assays. Our data show that Agamma-haplotypes represent genetic markers for the spectrum of thalassemic mutations, found in the Hellenic population and can constitute an important genetic repository upon which mutations leading to thalassemia and hemoglobinopathies occurred. PMID- 11421307 TI - Correspondence as conditional stimulus control: insights from experiments with pigeons. AB - Correspondence between saying and doing, typically studied in young children and individuals with developmental disabilities, was examined as an instance of conditional stimulus control. In Experiment 1, 3 pigeons were exposed to a two component repeated-trials procedure. In the first-sample or say-component, two response keys transilluminated by different colored lights were presented and the pigeon pecked one of the keys. After 1 s of darkness in the chamber, the second choice or do-component was presented, in which the two keys again were transilluminated, one by the color selected in the first component and the second by another color. Selecting the color that matched that selected in the say component resulted in access to food. Selecting the other color produced a blackout of the chamber. After an intertrial interval (ITI), the next say component was programmed, and the procedure was repeated. Correspondence remained at chance levels through several manipulations of ITI duration and sample response requirement. When a correction procedure was added such that only the originally selected sample stimulus was re-presented until a correct choice response occurred, reliable correspondence developed in 2 pigeons. This correspondence was eliminated by making reinforcement independent of correspondence and subsequently was reestablished when reinforcement again depended on correspondence. In Experiment 2, 3 other pigeons rapidly acquired correspondence under the final procedure used in Experiment 1. Increasing the time interval between the say and do components diminished correspondence. The results of the two experiments suggest how correspondence may be considered an instance of conditional stimulus control and that it is possible to construct a homologue of human say-do correspondence with pigeons. PMID- 11421308 TI - Assessing reinforcers under progressive schedule requirements. AB - Recent research findings suggest that reinforcing stimuli may be differentially effective as response requirements increase. We extended this line of research by evaluating responding under increasing schedule requirements via progressive ratio schedules and behavioral economic analyses. The differential effectiveness of preferred stimuli in treating destructive behavior maintained by automatic reinforcement also was examined. Results showed that one of two stimuli was associated with more responding under increasing schedule requirements for the 4 participants. Furthermore, stimuli associated with more responding under increasing schedule requirements generally were more effective in treating destructive behavior than stimuli associated with less responding. These data suggest that progressive-ratio schedules and behavioral economic analyses may be useful for developing a new technology for reinforcer identification. From a clinical perspective, these results suggest that two reinforcers may be similarly effective for low-effort tasks and differentially effective for high-effort tasks. PMID- 11421309 TI - A descriptive analysis of social consequences following problem behavior. AB - The social consequences delivered for problem behavior during functional analyses are presumed to represent common sources of reinforcement; however, the extent to which these consequences actually follow problem behavior in natural settings remains unclear. The purpose of this study was to determine whether access to attention, escape, or tangible items is frequently observed as a consequence of problem behavior under naturalistic conditions. Twenty-seven adults who lived in a state residential facility and who exhibited self-injurious behavior, aggression, or disruption participated. Observers recorded the occurrence of problem behavior by participants as well as a variety of consequences delivered by caregivers. Results indicated that attention was the most common consequence for problem behavior and that aggression was more likely to produce social consequences than were other forms of problem behavior. PMID- 11421310 TI - Longitudinal analysis of leisure-item preferences. AB - Twenty-two individuals with developmental disabilities participated in two leisure-item preference assessments, spaced approximately 16 months apart. Results showed (a) an overall increase in item contact across assessments for 13 participants and (b) at least some overlap across assessments in the five most highly ranked items for every participant. These results highlight individual differences in the stability of preference over time and suggest the need for research to identify the determinants of temporal shifts in preference. PMID- 11421311 TI - Increasing motorist compliance and caution at stop signs. AB - This study evaluated strategies to improve motorist compliance and caution at three stop-sign-controlled intersections with a history of motor vehicle crashes. The primary intervention was a light-emitting diode (LED) sign that featured animated eyes scanning left and right to prompt drivers to look left and right for approaching traffic. Data were scored from videotape on the percentage of drivers coming to a complete stop and the percentage of drivers looking right before entering the intersection. Observational data were collected on the percentage of right-angle conflicts (defined as braking suddenly or swerving from the path to avoid an intersection crash). The introduction of the LED sign according to a multiple baseline across the three intersections was associated with an increase in the percentage of vehicles coming to a complete stop at all three intersections and a small increase in the percentage of drivers looking right before entering the intersections. Conflicts between vehicles on the major and minor road were also reduced following the introduction of the animated eyes prompt. PMID- 11421312 TI - Functional analysis and treatment of self-injury associated with transitions. AB - We applied functional analysis methodology to the assessment and treatment of 2 individuals' self-injurious behavior (SIB), which was reported to be occasioned by transitions from one activity or location to another. A structural (task) analysis of activity transitions identified at least three separate components that might influence behavior either alone or in combination: (a) termination of a prechange activity, (b) initiation of a postchange activity, and (c) movement from one location to another. Results of preference and avoidance assessments were used to identify activities to which participants were exposed in varying arrangements during transitions in a functional analysis. Results of 1 participant's functional analysis indicated that his SIB was maintained by avoidance of having to change locations, regardless of the activity terminated prior to the change or the activity initiated following it. The 2nd participant's analysis revealed the same function but also an additional one: avoidance of certain task initiations. This information was used to identify transition contexts during intervention and to design treatment procedures appropriate for a given context and behavioral function. A procedure involving advance notice of an upcoming transition had no effect on SIB, and differential reinforcement of alternative behavior (DRA) had limited effects in the absence of extinction. Sustained decreases in SIB were observed when DRA was combined with extinction and response blocking. Further extensions of functional analysis methodology to the assessment of problem behavior in situations characterized by multiple or protracted stimulus changes are discussed. PMID- 11421313 TI - Behavioral economic influences on treatments designed to decrease destructive behavior. AB - In behavioral economics terms, response allocation is viewed as an exchange between the price of and the demand for reinforcers associated with various responses. In this study, behavioral economics principles were used to develop and evaluate a treatment package that reduced destructive behavior to zero while communication and compliance were increased. PMID- 11421314 TI - Habit reversal as a treatment for chronic skin picking in typically developing adult male siblings. AB - In this study, habit reversal was evaluated as a treatment for skin picking in typically developing adult male siblings using a nonconcurrent multiple baseline across participants design. Results showed socially valid decreases in reported picking as a result of treatment. PMID- 11421315 TI - Proximal and distal effects of play on child compliance with a brain-injured parent. AB - Individuals with brain injury may experience severe cognitive and other impairments. For brain-injured parents, such deficits may be associated with child behavior problems, including noncompliance. We assessed the effects of a play period conducted by a brain-injured father on the compliance of his son, who had become uncooperative with his father after the injury. The child consistently demonstrated improved compliance during proximal and distal compliance sessions that followed father-son play periods. PMID- 11421316 TI - Assessment and treatment of low-rate high-intensity problem behavior. AB - Functional analysis of low-rate aggression was conducted during extended observation periods and showed behavior to be maintained by positive reinforcement. Treatment consisted of variable-momentary differential reinforcement of other behavior and was successful in reducing problem behavior throughout these extended observation periods. PMID- 11421317 TI - Assessment and treatment of problem behavior maintained by escape from attention and access to tangible items. AB - The results obtained from two consecutive functional analyses conducted with a 6 year-old child with autism are described. In the initial functional analysis, the highest rates of problem behavior occurred in the play condition. In that condition, the delivery of attention appeared to occasion problem behaviors. A second functional analysis was conducted wherein an escape from attention condition and a tangible condition were added. In the second functional analysis, higher rates of responding were observed in the escape from attention and tangible conditions. The results suggested that problem behavior was maintained by negative reinforcement in the form of escape from attention and positive reinforcement in the form of gaining access to preferred tangible items. Problem behavior was treated using functional communication training combined with noncontingent reinforcement. PMID- 11421318 TI - Functional assessment of hand flapping in a general education classroom. AB - A functional assessment of hand flapping exhibited by a 5-year-old boy was conducted in a general education classroom. After a descriptive analysis ruled out several potential variables maintaining hand flapping, an experimental analysis was used to test the hypothesis that teacher-delivered task demands were functionally related to hand flapping. Results of the experimental analysis were used to develop a simple intervention for hand flapping. PMID- 11421319 TI - Antecedent manipulations in a tangible condition: effects of stimulus preference on aggression. AB - After a functional analysis indicated that aggression of an 8-year-old boy with autism was maintained by access to preferred items, antecedent manipulations involving the relative preference of restricted and noncontingently available stimuli were conducted. Restricting highly preferred items evoked the highest rates of aggression regardless of the preference level of the noncontingently available alternative items. Restricting less preferred stimuli was associated with moderate rates of aggression even when the alternative items were more preferred. PMID- 11421320 TI - Reinforcement contingencies and social reinforcement: some reciprocal relations between basic and applied research. AB - Reinforcement contingencies and social reinforcement are ubiquitous phenomena in applied behavior analysis. This discussion paper is divided into two sections. In the first section, reinforcement contingencies are discussed in terms of the necessary and sufficient conditions for reinforcement effects. Response-stimulus dependencies, conditional probabilities, and contiguity are discussed as possible mechanisms of, and arrangements for, reinforcement effects. In the second section, social reinforcement is discussed in terms of its functional subtypes and reinforcement context effects. Two underlying themes run throughout the discussion: (a) Applied research would benefit from a greater understanding of existing basic research, and (b) basic research could be designed to specifically address some of the issues about reinforcement that are central to effective application. PMID- 11421321 TI - Responses to heat stress in commercial and local broiler stocks. 1. Performance traits. AB - 1. This study investigated effects of conditioning at 5 d of age and food restriction during a hot period on responses to prolonged heat stress of two fast (G1 and G2) and one slower-growing (G3) broiler stocks. 2. Chicks from each stock were divided into 3 groups: control, conditioned (chicks exposed to 36 degrees C for 24 h at 5 d of age) and food restricted (during the heat stress food was withdrawn 2 h before the hot period and chicks were fed between 17.00 and 08.00 h). Prolonged heat stress was induced daily from 28 to 49 d by heating until the ambient temperature reached 32 degrees to 35 degrees C between 10.00 and 17.00 h. 3. Body weights, mortality and rectal temperatures were lower and food conversion higher for stock G3 than G1 or G2. 4. Management procedures used in this study improved food conversion and lowered mortality without affecting body weight. Rectal temperatures of the treated groups were similar to those of the controls. PMID- 11421322 TI - Responses to heat stress in commercial and local broiler stocks. 2. Developmental stability of bilateral traits. AB - 1. Developmental stability of bilaterally symmetrical traits was measured in two fast (G1 and G2) and one slower growing (G3) broiler stocks when conditioned (chicks exposed to 36 degrees C for 24 h at 5 d of age) or food restricted (food withdrawn 2 h before the hot period and chicks fed between 17.00 and 08.00 h) during heat stress administered daily from 28 to 49 d of age. 2. Length of face, length, width and weight of shanks, weights of P. major and P. minor muscles, and weights of lung and caeca were chosen as bilateral traits representative of skeletal, muscle, respiratory and digestive systems. 3. Development of skeletal bilateral traits was affected primarily by genotype and sex. The heart:lung ratio decreased with conditioning in stocks G1 and G2 at 21 d. Face length, shank length and shank weight exhibited antisymmetry, whereas shank width and weights of P. minor, lung and caeca exhibited fluctuating asymmetry and antisymmetry. For P. major weight, there was directional asymmetry. 4. In general, relative asymmetry of skeletal traits was lower than for traits associated with muscle and with respiratory and digestive systems. Relative asymmetry of P. major weight was lower in conditioned and G3 broilers than in their respective counterparts. 5. If relative asymmetry is a valid indicator, item heat stress may be less in slower growing and conditioned broilers. PMID- 11421323 TI - Influence of environmental enrichment on injurious pecking and perching behaviour in young turkeys. AB - 1. In order to reduce injurious pecking, the influence of environmental enrichment on pecking and perching behaviour was studied in young male and female turkeys. 2. Two different types of enrichment and a control treatment (TO) were tested: T1, metal objects and straw; T2, similar to T1 + wood perches. Birds were housed in 36 m2 pens at a light intensity of 5 lux with 4 replicates per treatment in a 2x3 factorial design. 3. Pecking at objects and perching behaviour were observed weekly. Behaviour was video recorded at weeks 5 and 10. Birds were examined daily for the occurrence of injuries. At the end of the rearing period, an ACTH challenge was performed and H/L ratio was measured. 4. Objects were regularly pecked at. Perching was more common in females, peaked at week 5 (10% to 13% of birds perched) and declined to 0% by week 10. Aggressive pecking was more frequently observed in males in T0 than in T1 or T2 at week 10. Wing (in males and females), tail and head (in males) injuries were more common in T0 than in T1 or T2. T1 and T2 were similar Response to ACTH challenge and H/L ratio were not consistently influenced by treatments. 5. It is concluded that metal objects and straw reduced injurious pecking in young female and male turkeys by redirecting, pecking activity. PMID- 11421324 TI - Release of heat, moisture and carbon dioxide in an aviary system for laying hens. AB - 1. Production of total heat, divided into sensible and latent heat, together with carbon dioxide and animal activity were determined at different ambient temperatures under full-scale conditions in an aviary system with loose-housed laying hens. 2. Sensible heat production decreased approximately linearly with increasing ambient temperature and was lower during the day than at night. One explanation may be that some sensible heat produced by the hens was converted to latent heat by evaporation of moisture due to increased activity of the hens during the day (scratching in the bedding and drinking/waste water). 3. Latent heat production increased with increasing ambient temperature and was higher during the day than at night. This confirms that the hens, by agitating the bedding during the day and by spilling drinking water, transferred some sensible heat to latent heat by evaporation. 4. Total heat production decreased with increasing temperature because the hens by thermoregulation decreased their metabolism in order to maintain a constant body temperature. The difference between day and night values of total heat production was less pronounced than in the case of sensible and latent heat. In comparison with current guidelines the measurements showed a higher total heat production (22% higher at 20 degrees C). 5. There was a large diurnal variation in carbon dioxide production, closely correlated to measured animal activity; on average carbon dioxide production during the 12-h dark period was only 66% of the production during the day. PMID- 11421325 TI - Large granular lymphocytes in the oviduct of developing and hormone-treated chickens. AB - 1. In the present study the distribution of large granular lymphocytes (LGLs) and influence of oestrogen on these cells was investigated in the oviduct of chickens by the observation of H and E stained sections. 2. The LGLs first appeared in the oviduct at 9 weeks and their frequency of occurrence increased with age and was highest in the infundibulum and vagina from 21 to 32 weeks of age. The frequency of occurrence of LGLs from magnum to uterus (glandular part) was low. These cells were located at the base, middle and apical parts of the lining epithelium as well as in the sperm-storage glandular epithelium. 3. When chickens of 21 weeks were treated with DES, LGLs abundantly infiltrated in the oviducal epithelium in comparison to the control. 4. These results suggested that the development of LGLs in the oviduct of chickens correlates with oestrogen secretion. PMID- 11421326 TI - Effects of strain, maternal age and sex on morphological characteristics and composition of tibial bone in broilers. AB - 1. This study was to determine the effects of strain, age of the maternal flock and sex on morphological characteristics and composition of tibial bone of broilers from hatch to 48 d of age. 2. A total of 600 chicks was obtained from 2 strains of broiler breeder flocks (150/chicks/strain/maternal age). Maternal flock age was classified as young (32 to 35 weeks of age) or old (56 to 58 weeks of age). Birds were reared under standard feeding and lighting regimes. 3. On day 1, 16, 32 and 48, twelve birds were selected at random from each maternal group, strain and sex and killed. The wet bone weight and volume were measured. Morphological characteristics of tibia were determined using radiography. Bone breaking strength was tested. Tibia dry matter, ash content, mineral density and collagen level were determined. 4. A quadratic increase occurred with increase in age of broilers for all variables, except proximal width, medial cortex thickness and distal condyle width which increased in a linear manner. 5. Maternal age had a significant effect only on the variably measured at the time of hatch. On day of hatch bone weight, ash content and bone volume were affected by maternal age, but the extent of this also depended on the strain. 6. The differences observed between strains for bone anatomy and bone mineralisation during the rapid growth period of 16 d were not significant at later ages, with the exception of bone volume. Differences between sexes were evident from 16 to 49 d of age with females having lower values. PMID- 11421327 TI - Monthly model for genetic evaluation of laying hens 1. Fixed regression. AB - 1. This paper addresses the possibility of using a monthly model for the genetic evaluation of laying hens, based on the definition of a test day model with fixed regression as used in dairy cattle, in which monthly records were treated as repeated measurements of the same trait. 2. Production records of 6450 hens, daughters of 180 sires and 1335 dams were analysed using an animal model with restricted maximum likelihood (REML). The traits considered were individual monthly egg production and cumulative egg production in 11 months. Four different models were fitted to various combinations of monthly and cumulative records. The covariates were derived from the regression of Ali and Schaeffer (1987). 3. Spearman rank correlations were computed to compare breeding values from different models. Two types of correlations were computed: between individual breeding values and between sire breeding values based on subsets of full-sib records. 4. The results indicated that a monthly model with nested covariates produced higher heritability and permanent environmental variance than the models with non-nested or without covariates. The estimates of heritability obtained from monthly model were lower than the estimates from the cumulative model. The monthly model resulted in higher correlations of sire breeding values between two subsets of full-sib records than those from cumulative models. 5. In conclusion, the monthly model with nested covariates appears to be better than the model with non-nested covariates or without covariate. Although the heritability estimates obtained from the monthly model were lower, the monthly model with nested covariates could be better than the cumulative model for genetic evaluation of laying hens in the 1st cycle of laying period when using either full or part records. The use of information from odd months of production could be of interest for the evaluation of full records. PMID- 11421328 TI - Effects of dietary spirulina on meat colour in muscle of broiler chickens. AB - 1. The present study was undertaken to determine the effects of dietary spirulina on growth performance and pigmentation in the muscle of growing broiler chickens and to examine the possibility that zeaxanthin in spirulina may affect yellow colour development in the meat. 2. Twenty-four, 21-d-old, male broiler chicks were fed an experimental diet containing spirulina at 0, 40, or 80 g/ kg for 16 d. No significant differences among treatments were observed in body weights, nor weights or yields (as a percentage of body weight) for any of the selected traits, including liver, abdominal fat, kidney and Pectoralis profundus. 3. Spectrocolourimetric analyses revealed that the redness of Pectoralis superficialis, profundus and Sartorius muscles reached a maximum in chicks fed the 40 g/kg spirulina diet, while the yellowness of all fillets, including the Semitendinosus muscle, increased in a sub-linear fashion with increased spirulina in the diet. The overall correlation between the yellowness and zeaxanthin content in the Pectoralis muscle was significant. 4. This study provides the first conclusive evidence that dietary spirulina influences both the yellowness and redness of broiler flesh and that the increments in yellowness with dietary spirulina content may possibly be reflected in the common yellow pigment related to the accumulation of zeaxanthin within the flesh. PMID- 11421329 TI - The influence of pelleted feed on the response of growing pullets to photoperiods of less than ten hours. AB - 1. An experiment was designed to test the hypothesis that delayed sexual maturity in pullets reared on very short (4 h) constant photoperiods might be partly attributable to limitation of food intake and that offering a pelleted feed might circumvent this effect. 2. The factors investigated were 2 strains (Amber Link and Hyline Brown), 3 photoperiods (4, 7 and 10 h) and 2 forms of food (mash throughout rearing or crumbs from 0 to 4 weeks followed by pellets). All 12 combinations of these factors were tested with 14 replications of 18 pullet chicks allocated to each combination. 3. Mean ages at first egg for 4, 7 and 10 h rearing photoperiods were 189, 184 and 162 d respectively. Pullets given the pelleted diet ate 2% less food to 20 weeks but were 6% heavier at that age. However, the pellet-fed birds were 6 d later in mean age at 50% lay. There was no interaction between form of food and photoperiod in the data for age at first egg. 4. It is concluded that constant short photoperiods during rearing cause delayed sexual maturity entirely due to the effect of light on gonadal development and that limitation of food intake is not a factor in this response. PMID- 11421330 TI - The effect of dietary chloride and bicarbonate on blood pH, haematological variables, pulmonary hypertension and ascites in broiler chickens. AB - 1. The effect of supplementing grower diets with bicarbonate or chloride on haematological variables, pulmonary hypertension syndrome and ascites in broilers exposed to cold temperature was investigated. 2. High concentrations of dietary chloride had no effect on the pH of the venous blood but a low chloride/high bicarbonate diet significantly increased blood pH. There was no consistent effect of dietary chloride or bicarbonate concentrations on growth performance, although in 1 experiment birds given a low chloride/high bicarbonate diet consumed less food and gained less weight than controls. 3. Birds fed on high-chloride diets tended to have a higher incidence of ascites and pulmonary hypertension than controls. Birds fed on low-chloride and high-bicarbonate diets had significantly lower pulmonary hypertension and lower heart weights, which may have indicated a decrease in pulmonary and systemic blood pressure. 4. We conclude that increasing dietary bicarbonate and reducing dietary chloride has potential as a low cost and effective method to reduce the pulmonary hypertension which leads to ascites in broiler chickens. PMID- 11421331 TI - Nitrogen retention and performance of brown laying hens on diets with different protein content and constant concentration of amino acids and energy. AB - 1. The aim of this study was to determine the nitrogen balance and the performance of laying hens fed on diets with a protein content lower than the diets currently used in commercial practice but with adequate concentrations of lysine, sulphur amino acids, tryptophan and threonine. 2. Ninety-six Hy-Line Brown hens, 24 weeks old, were divided into 3 groups of 8 replicates and received, for 16 weeks, diets formulated to have 3 different protein concentrations: 170 (control), 150 and 130 g/kg CP and the same energy content. For each protein concentration, the contents of lysine, methionine, methionine+cystine, tryptophan and threonine were maintained at minimum requirement concentrations by supplying synthetic amino acids. 3. In the first half of the trial, egg production and egg weight were similar in all groups. From the 9th week onwards group 150 CP laid heavier eggs and had a slightly lower egg deposition and total mass. Food conversion ratio was best in the control group. 4. Nitrogen intake was related to the protein concentration of the diet, the food intake being almost the same in the 3 experimental groups. Faecal nitrogen content significantly and linearly decreased with reduction in dietary protein content and was about 50% of the intake. Considering the nitrogen faecal/intake ratio, the 150 CP group showed better nitrogen utilisation at each sampling time. PMID- 11421332 TI - Effects of preservation method and beta-glucanase supplementation on ileal amino acid digestibility and feeding value of barley for poultry. AB - 1. Effects of preservation method (drying or air-tight storage of whole grain and ensiling of rolled high-moisture grain) and beta-glucanase supplementation (Econase) on apparent ileal amino acid digestibilities and metabolisable energy content of barley were evaluated with Ross broiler chickens. In addition, the effect of barley preservation method was assessed using Leghorn cockerels. 2. Birds were given either a semi-purified soyabean meal basal diet or a mixture of the basal diet and barley (50:50 on dry matter basis). Apparent ileal digestibilities (AID) of nutrients were assessed using the slaughter technique. AID of nutrients and nutrient digestibility measured using excreta (AED) were determined using chromium mordanted straw as an indigestible marker. 3. In broilers, AID of amino acids, dry matter and organic matter were lower for dried than air-tight stored barley, particularly for diets based on ensiled barley. In cockerels, barley preservation method had no effect on amino acid AID. The AED of nutrients and nitrogen corrected apparent metabolisable energy content (AMEn) was highest for ensiled barley across both experiments. 4. beta-glucanase supplementation increased nutrient digestibility, phosphorus retention and AMEn content of air-tight stored and dried barley diets in particular but had only negligible effects on ensiled barley. Beta-glucanase improved the AID of amino acids in dried barley but not in air-tight stored or ensiled barley. 5. Amino acid digestibilities were lower in broilers than cockerels and the effect of barley preservation on feeding value of barley was different for broilers and cockerels. PMID- 11421333 TI - The effects of dietary L-carnitine supplementation on the performance, organ weights and circulating hormone and metabolite concentrations of broiler chickens reared under a normal or low temperature schedule. AB - 1. The present study examined the effects of the addition of 100 mg/kg L carnitine to the basal starter (containing 17.8 mg/kg L-carnitine) and finisher (containing 22.9 mg/kg L-carnitine) diets on performance, organ weights and plasma hormone and metabolite concentrations of male and female broiler chickens. The broiler chickens were reared either in a room with a normal temperature (NT) program or with a low temperature (LT) program (rapid decrease from 28 degrees C to 20 degrees C at 14 d of age). 2. Broiler chickens reared under the LT schedule consumed more food and attained a greater body weight at 42 d of age than their counterparts reared under NT schedule, without any difference in food efficiency or abdominal fat content. Dietary L-carnitine supplementation had no significant effects on any of these production parameters, except for a reduction in the abdominal fat content of female NT chickens. However, the LT schedule and dietary L-carnitine supplementation greatly increased absolute and proportional heart weights. The elevated heart weights were not due to right ventricle hypertrophy. 3. Both the LT program and L-carnitine supplementation increased circulating plasma triiodothyronine concentrations. There were also some transient effects of both experimental variables on plasma growth hormone, glucose and triglyceride concentrations. 4. L-carnitine did not improve broiler performance. However, this result does not mean that L-carnitine supplementation cannot have beneficial effects in other circumstances. In view of the elevated proportional heart weights, it can also be argued that L-carnitine is a potential agent for reducing the incidence of metabolic diseases in broiler chickens. PMID- 11421334 TI - Nutritional value of raw and autoclaved kabuli and desi chickpeas (Cicer arietinum L.) for growing chickens. AB - 1. Two experiments were carried out to determine the effect of inclusion of raw (kabuli and desi) and autoclaved (desi) chickpea seeds in wheat-based starter diets in chickens grown to 28 d of age on the performance, digestive organ sizes, nitrogen-corrected apparent metabolisable energy (AMEn), ileal apparent protein and starch digestibilities (APD and ASD) and intestinal alpha-amylase and trypsin activities. 2. In the first experiment, diets were formulated to contain 0, 150, 300 and 450 g/kg of raw kabuli chickpea seeds. Increasing the proportion of seed in the diet negatively influenced body weight gain, food intake and food efficiency. The relative weights of the pancreas, liver and gizzard and the relative lengths of duodenum, jejunum, ileum and caeca were increased significantly when the chickpea seeds were included in the diets. Correspondingly, APD, ASD, alpha-amylase and trypsin activities and AMEn were reduced significantly when the chickpea seed was incorporated in the diets. 3. In the 2nd experiment, diets were formulated to contain 75 and 150 g/kg of raw and autoclaved desi chickpea seeds. Weight gain and food intake of the chicks given desi chickpea diets were significantly reduced compared with those fed on the control diet. Increasing the proportion of seed in the diet negatively influenced body weight gain, food intake and food efficiency. Moreover, a significant increment in the relative weights of liver and pancreas, and in the relative lengths of duodenum, ileum and caeca was observed when the concentration of chickpea seeds in the diets was increased. Feeding autoclaved seeds significantly increased the weight gains and the food intakes. However, food efficiency was not modified by the autoclaving. Relative weights of gizzard and liver and relative lengths of ileum were decreased significantly by the inclusion of autoclaved desi chickpea in the diet. 4. We concluded that the inclusion of kabuli (up to 450 g/kg) and desi (up to 150 g/kg) chickpea seeds produced a negative effect on the performance of the birds, and an increment in the relative weights and lengths of the digestive organs. In addition, the incorporation of kabuli chickpea produced a reduction of protein and starch digestibilities, alpha-amylase and trypsin activities, and AMEn of food compared with the birds given the control diet. Autoclaved treatment of desi chickpea improved the performance of the birds. PMID- 11421335 TI - Comparative evaluation of the effect of two maternal diets on fatty acids, vitamin E and carotenoids in the chick embryo. AB - 1. The fatty acid profile of egg yolk and vitamin E and carotenoid accumulation in the egg yolk and embryonic tissues were investigated in relation to the maternal diet. 2. Two hundred fertile eggs (Ross 308 Broiler Breeder), obtained from hens fed on a maize-based (M-diet) or a wheat-based diet (W-diet), were incubated using standard conditions. 3. The egg yolk and embryo tissues (residual yolk, yolk sac membrane, liver, kidney, lung, muscles, adipose tissue and plasma) were collected on d 18 of incubation and on d 21 (newly-hatched chicks) and analysed for fatty acids, vitamin E and carotenoids. 4. The diets did not differ in terms of fatty acid or alpha-tocopherol concentrations. The concentration of carotenoids in the M-diet was 11.8 mg/kg and in the W-diet was 5.6 mg/kg with lutein and zeaxanthin being major carotenoids. 5. Eggs from the M-group contained higher (P<0.01) concentrations of beta+gamma-tocopherols, total carotenoids, lutein and zeaxanthin. Chickens hatched from those eggs were characterised by the increased concentrations of total carotenoids and zeaxanthin in all the tissues studied. The concentration of beta+gamma-tocopherol was enhanced only in the liver and yolk sac membrane. 6. It is concluded that the maternal diet plays an important role in antioxidant systems formation during chick embryonic development; the M-diet can increase the antioxidant potential of the egg yolk and embryonic tissues compared to the antioxidant potential provided by parent birds fed the W-diet. PMID- 11421336 TI - Sequential studies of skeletal calcium reserves and structural bone volume in a commercial layer flock. AB - 1. Production-induced osteoporosis in caged laying hens is thought to represent a major constraint to continued genetic development. 2. The relationship between body weight, egg production, skeletal abnormalities characteristic of osteoporosis, femur calcium and bone histology was examined in a flock of ISA Brown layers from 16 to 68 weeks of age. 3. Experiment 2 examined a flock of Lohmann browns for skeletal abnormalities characteristic of osteoporosis at 45 weeks of age and the severity of abnormalities was then related to body weight and production between 18 and 45 weeks of age. 4. Average body weight declined in the ISA flock between 35 and 45 weeks of age, which correlated with a loss of skeletal calcium reserves (15% to 20%) and with the induction of osteoporosis. Between 42 and 68 weeks of age, birds were able to replenish femur calcium levels. 5. Birds in the Lohmann flock showing severe skeletal abnormalities at 45 weeks of age experienced weight loss between 27 and 31 weeks of age, which was associated with a decrease in egg production of 18%. After 35 weeks of age, egg production of these birds recovered to similar levels as unaffected or mildly affected birds. 6. It seems likely that better standardisation of the equilibrium between growth, skeletal reserves, food intake and egg production can reduce osteoporosis, as well as improving the productive potential of modern laying strains. PMID- 11421337 TI - Sperm-egg penetration in laying breeder flocks: a technique for the prediction of fertility. AB - 1. An experiment was conducted to evaluate indices of fertility including the sperm penetration (SP) assay as a technique for the prediction of fertility. Forty-eight males consisting of White Leghorn (WL), New Hampshire (NH), Iraqi Brown (IBr) and Iraqi Barred (IBa) (12 males each) and 64 WL hens were divided at random into 4 groups of 4 replicates of 3 males and 4 females each. 2. At the beginning of each week semen was collected from males and pooled by breed of male. Hens in each breeding group were inseminated once weekly, by breeding group, for 4 consecutive weeks with pooled semen from WL, NH, IBr and IBa males (WLxWL, NHxWL, IBrxWL and IBaxWL). 3. The differences in percentage of dead sperm, acrosomal abnormalities, mass motility, individual motility and spermatocrit between the experimental breeds demonstrated the superiority of WL and NH males in all these quantitative characters of the semen. On the other hand, WL hens inseminated with spermatozoa from NH males had significantly more sperm-egg penetration (SP) holes than WL hens inseminated with spermatozoa from other breeds of males. The breed of males used for insemination affected fertility, hatchability and embryonic mortality. 4. The highest fertility and hatchability and lowest embryonic mortality were observed in eggs laid by hens inseminated with spermatozoa from WL and NH males in comparison with hens inseminated with spermatozoa from Iraqi males. 5. There was a strong positive correlation between SP values and fertility for WLxWL, NHxWL, IBrxWL and IBaxWL. The correlation for all breeds combined was also significant. In addition, SP was also positively correlated with hatchability and negatively correlated with embryonic mortality. PMID- 11421338 TI - Influence of hydration on the mechanical performance of duck down feathers. AB - 1. We report the first measurements to be made on the effects of hydration on the mechanical properties of commercially-prepared duck down feathers. Tensile tests were performed, to failure, on isolated barbs of down feathers at 50% RH and with fully-hydrated (wet) feathers. 2. The Young's modulus (stiffness) and ultimate stress (strength) of the feathers was higher at 50% RH than when wet. Wet feathers, however, failed at greater extension than dry feathers. The implications for down feather performance and for the use of feathers as a structural material are discussed. PMID- 11421339 TI - Inheritance of the black hackle of the Indian Game bantam. AB - 1. An investigation was conducted among the progeny from crosses between pea combed Indian Game and single combed double-laced Barnevelder bantams in an effort to determine the inheritance of the black hackle phenotype of the former. 2. The double-laced feather pattern phenotype of both breeds has been shown to depend upon homozygosity of both the linked eumelanin extension melanotic ML*M and the feather pattern arranging gene PG*P. However the hackles of the Indian Game are black, whilst those of the Barnevelder are partially striped, which suggests the presence of an extra eumelaniser in the genome of the Indian Game. This could not be due to variation at the extended black-E*-locus since the Indian Game and the Barnevelder have been shown respectively to depend upon the wheaten E*WH and brown E*B alleles; the former normally less heavily eumelanised. 3. A mating of a Barnevelder male with an Indian Game female produced 11 chicks, all of which were black in hackle. The mating of 2 F1 males with 5 Barnevelder females produced 35 chicks in the backcross to the double recessive, all of which were of parental phenotype. These comprised 18 peacomb, black in hackle and 17 single comb, partially striped hackle; classification being clear injuvenile plumage. 4. The absence of crossovers in the backcross suggest allelism or extremely close linkage between the loci of peacomb and the dominant eumelanin extension which causes the black in hackles. 5. The eumelanin extension isolated in this work appears therefore to be allelic to, if not a further manifestation of the well-established charcoal CHA*C gene. PMID- 11421340 TI - Discrimination between individual humans by domestic fowl (Gallus gallus domesticus). AB - 1. We demonstrate that Barred-Rock and Isa-Brown hens can discriminate between the presence of 2 different humans and use this information as a cue for whether or not to make an operant response. 2. This demonstration in domestic fowl, the first of its kind in any avian subject, is consistent with a growing body of evidence that many species, including agricultural animals, are surprisingly adept at differentiating between humans. 3. Such a discriminative ability carries considerable implications for issues related to husbandry, production and welfare. PMID- 11421341 TI - An appreciation of Elvin A. Kabat (1914-2000): scientist, educator and a founder of modern carbohydrate biology. PMID- 11421342 TI - A journey to the world of glycobiology. AB - Finding of the deletion phenomenon of certain oligosaccharides in human milk and its correlation to the blood types of the donors opened a way to elucidate the biochemical basis of blood types in man. This success led to the idea of establishing reliable techniques to elucidate the structures and functions of the N-linked sugar chains of glycoproteins. N-Linked sugar chains were first released quantitatively as oligosaccharides by enzymatic and chemical means, and labelled by reduction with NaB3H4. After fractionation, structures of the radioactive oligosaccharides were determined by a series of methods developed for the studies of milk oligosaccharides. By using such techniques, structural rules hidden in the N-linked sugar chains, and organ- and species-specific N-glycosylation of glycoproteins, which afforded a firm basis to the development of glycobiology, were elucidated. Finding of galactose deficiency in the N-linked sugar chains of serum IgG from patients with rheumatoid arthritis, and malignant alteration of N glycosylation in various tumors opened a new research world called glycopathology. However, recent studies revealed that several structural exceptions occur in the sugar chains of particular glycoproteins. Finding of the occurrence of the Galbeta1-4Fucalpha1- group linked at the C-6 position of the proximal N-acetylglucosamine residue of the hybrid type sugar chains of octopus rhodopsin is one of such examples. This finding indicated that the fucosyl residue of the fucosylated trimannosyl core should no more be considered as a stop signal as has long been believed. Furthermore, recent studies on dystroglycan revealed that the sugar chains, which do not fall into the current classification of N and O-linked sugar chains, are essential for the expression of the functional role of this glycoprotein. It was found that expression of many glycoproteins is altered by aging. Among the alterations of the glycoprotein patterns found in the brain nervous system, the most prominent evidence was found in P0. This protein is produced in non-glycosylated form in the spinal cord of young mammals. However, it starts to be N-glycosylated in the spinal cord of aged animals. These evidences indicate that various unusual sugar chains occur as minor components in mammals, and play important roles in particular tissues. PMID- 11421343 TI - The joys of HexNAc. The synthesis and function of N- and O-glycan branches. AB - This review covers discoveries made over the past 30-35 years that were important to our understanding of the synthetic pathway required for initiation of the antennae or branches on complex N-glycans and O-glycans. The review deals primarily with the author's contributions but the relevant work of other laboratories is also discussed. The focus of the review is almost entirely on the glycosyltransferases involved in the process. The following topics are discussed. (1) The localization of the synthesis of complex N-glycan antennae to the Golgi apparatus. (2) The "evolutionary boundary" at the stage in N-glycan processing where there is a change from oligomannose to complex N-glycans; this switch correlates with the appearance of multicellular organisms. (3) The discovery of the three enzymes which play a key role in this switch, N acetylglucosaminyltransferases I and II and mannosidase II. (4) The "yellow brick road" which leads from oligomannose to highly branched complex N-glycans with emphasis on the enzymes involved in the process and the factors which control the routes of synthesis. (5) A short discussion of the characteristics of the enzymes involved and of the genes that encode them. (6) The role of complex N-glycans in mammalian and Caenorhabditis elegans development. (7) The crystal structure of N acetylglucosaminyltransferase I. (8) The discovery of the enzymes which synthesize O-glycan cores 1, 2, 3 and 4 and their elongation. PMID- 11421344 TI - Achievements and challenges of sialic acid research. AB - Slalic acids are one of the most important molecules of life, since they occupy the terminal position on macromolecules and cell membranes and are involved in many biological and pathological phenomena. The structures of sialic acids, comprising a family of over 40 neuraminic acid derivatives, have been elucidated. However, many aspects of the regulation of their metabolism at the enzyme and gene levels, as well as of their functions remain mysterious. Sialic acids play a dual role, not only are they indispensable for the protection to and adaptation of life, but are also utilised by life-threatening infectious microorganisms. In this article the present state of knowledge in sialobiology, with an emphasis on my personal experience in this research area, is outlined including a discussion of necessary future work in this fascinating field of cell biology. PMID- 11421345 TI - Unravelling the biochemical basis of blood group ABO and Lewis antigenic specificity. AB - The ABO blood-group polymorphism is still the most clinically important system in blood transfusion practice. The groups were discovered in 1900 and the genes at the ABO locus were cloned nearly a century later in 1990. To enable this goal to be reached intensive studies were carried out in the intervening years on the serology, genetics, inheritance and biochemistry of the antigens belonging to this system. This article describes biochemical genetic investigations on ABO and the related Lewis antigens starting from the time in the 1940s when serological and classical genetical studies had established the immunological basis and mode of inheritance of the antigens but practically nothing was known about their chemical structure. Essential steps were the definition of H as the product of a genetic system Hh independent of ABO, and the establishment of the precursor product relationship of H to A and B antigens. Indirect methods gave first indications that the specificity of antigens resided in carbohydrate and revealed the immunodominant sugars in the antigenic structures. Subsequently chemical fragmentation procedures enabled the complete determinant structures to be established. Degradation experiments with glycosidases revealed how loss of one specificity by the removal of a single sugar unit exposed a new specificity and suggested that biosynthesis proceeded by a reversal of this process whereby the oligosaccharide structures were built up by the sequential addition of sugar units. Hence, the primary blood-group gene products were predicted to be glycosyltransferase enzymes that added the last sugar to complete the determinant structures. Identification of these enzymes gave new genetic markers and eventually purification of the blood-group A-gene encoded N acetylgalactosaminyltransferase gave a probe for cloning the ABO locus. Blood group ABO genotyping by DNA methods has now become a practical possibility. PMID- 11421346 TI - The chemistry and immunochemistry of blood group A, B, H, and Lewis antigens: past, present and future. AB - This article traces reseach on the chemistry and immunochemistry of blood group A, B, H, and Lewis antigens from early work on the identification of soluble sources of these antigens, through the elucidation of the structures of the carbohydrate epitopes responsible for these specificities, to recent work on exploring their possible use as cancer vaccines. The various approaches used in the isolation of oligosaccharides from mucins for use in structural studies are discussed, as are recent efforts in the chemical systhesis of blood group-active oligosaccharides. PMID- 11421347 TI - Affinity enhancement by multivalent lectin-carbohydrate interaction. AB - The binding of simple carbohydrate ligands by proteins often requires affinity enhancement to attain biologically relevant strength. This is especially true for endocytotic receptors and the molecules that engage in the first-line of defense. For such purposes, nature often utilizes a mode of affinity enhancement that arises from multiple interactions between the binding proteins and the carbohydrate ligands, which we term glycoside cluster effect. In this review article we give a number of examples and describe important factors in the multi valent interactions that govern the degree of affinity enhancement. PMID- 11421348 TI - Progress in deciphering the information content of the 'glycome'--a crescendo in the closing years of the millennium. AB - The closing years of the second millennium have been uplifting for carbohydrate biology. Optimism that oligosaccharide sequences are bearers of crucial biological information has been borne out by the constellation of efforts of carbohydrate chemists, biochemists, immunochemists, and cell- and molecular biologists. The direct involvement of specific oligosaccharide sequences in protein targeting and folding, and in mechanisms of infection, inflammation and immunity is now unquestioned. With the emergence of families of proteins with carbohydrate-binding activities, assignments of information content for defined oligosaccharide sequences will become more common, but the pinpointing and elucidation of the bioactive domains on oligosaccharides will continue to pose challenges even to the most experienced carbohydrate biologists. The neoglycolipid technology incorporates some of the key requirements for this challenge: namely the resolution of complex glycan mixtures, and ligand binding coupled with sequence determination by mass spectrometry. PMID- 11421349 TI - So what do your sugars do? AB - Up to about fifteen years ago the above question was quite often asked of glycoconjugates enthusiasts. It was always a silly question, the answer being 'How many doings do you want to hear about?' Since then our understanding of the structures, biosynthesis and varied functions of the glycoconjugates has developed incredibly and come to have a major impact on much of present-day biological and medical research. PMID- 11421351 TI - 'Heparin'--from anticoagulant drug into the new biology. AB - This overview attempts to cover, from a personal viewpoint, the development of the 'heparin' field during the last four decades. In particular, it emphasizes the metamorphosis of heparan sulfate (HS), from a disturbing contaminant in heparin production to the present-day key player in cell and developmental biology. Our understanding of the structural properties of the polysaccharides has been greatly promoted by studies of their biosynthesis. We now have a fairly detailed view of the various enzymatic reactions, that convert the initial [4GlcAbeta1-4GlcNAcalpha1-]n polymer into sulfated products with highly variable proportions of GlcA/IdoA and of N-acetyl, N-sulfate and O-sulfate substituents. It is also recognized that the variously substituted domains of the polysaccharide serve to interact, in more or less specific fashion, with a multitude of proteins, and that these interactions are essential to the biological functions of the proteins. Molecular genetics has unravelled the gene structures for almost all of the enzymes required to synthesize a heparin or HS chain, and has shown that several of these proteins exhibit genetic polymorphism. While differences in substrate specificity between enzyme isoforms may help to explain the structural variability of, in particular, HS chains, we still only partly understand the key features of heparin/HS biosynthesis and its regulation. PMID- 11421350 TI - Protein-bound carbohydrates on cell-surface as targets of recognition: an odyssey in understanding them. AB - Multidisciplinary approaches by a number of investigators have established that cell-surface carbohydrates are integral components of recognition systems regulating survival, migration, adhesion, growth and differentiation of various cells. Our own experience and contributions to this exciting field are described. We discovered Endo D as the first endoglycosidase acting on glycoproteins, found complementary specificity of two endoglycosidases (Endo D and Endo H), and applied these enzymes for glycoprotein research. Endo-beta-galactosidase C, which hydrolyzes Galalpha1-3Galbeta1-4GlcNAc xenoantigenic determinant, was later found and molecularly cloned. We also found highly branched poly-N-acetyllactosamines in early embryonic cells, and demonstrated developmentally regulated carbohydrate changes during early mammalian development. The binding site for Dolichos biflorus agglutinin was introduced as a new differentiation marker. Basigin and embigin, two related members of the immunoglobulin superfamily, a sialomucin MGC 24 and other glycoproteins were discovered as carriers of developmentally regulated carbohydrate markers. We proposed enhancement of integrin action as a function of sugar chains with Lewis X epitope, and observed a relationship between the expression of carbohydrate markers and invasive properties of human carcinoma. Midkine, a heparin-binding growth factor, was discovered more recently and its interaction with heparin and oversulfated chondroitin sulfate was elucidated. N-Acetylglucosamine-6-sulfotransferase was cloned and used to reconstitute L-selectin ligands. Gene knockout was applied to reveal in vivo function of basigin, syndecan-4 and chondroitin 6-sulfate. Throughout my research on all these subjects, I have been fortunate in obtaining unexpected observations and enjoying fruitful collaborations. PMID- 11421352 TI - Hyaluronan, a common thread. AB - Hyaluronan, nature's simplest, but still exceptionally versatile glycosaminoglycan, is currently the focus of attention across a wide front of research; from cell biology, morphogenesis, matrix organization, pathobiology to tissue engineering. This macromolecule has entangled me in a number of puzzling and challenging projects over the past 3 decades. These entertaining encounters are outlined in this retrospective. PMID- 11421353 TI - Terminal glycosylation and disease: influence on cancer and cystic fibrosis. AB - Terminal glycosylation has been a recurring theme of the laboratory. In cystic fibrosis (CF), decreased sialic acid and increased fucosyl residues in alpha1,3 position to antennary N-acetyl glucosamine is the CF glycosylation phenotype. The glycosylation phenotype is reversed by transfection of CF airway cells with wtCFTR. In neuronal cells, polymers of alpha2,8sialyl residues are prominent in oligodendrocytes and human neuroblastoma. These findings are discussed in relationship to early studies in our laboratories and those of other investigators. The potential extension of these concepts to future clinical therapeutics is presented. PMID- 11421354 TI - Traveling for the glycosphingolipid path. AB - Our studies on glycosphingolipids (GSLs) were initiated through isolation and structural characterization of lacto-series type 1 and 2 GSLs, and globo-series GSLs. Lacto-series structures included histo-blood group ABH and I/i antigens. Our subsequent studies were focused on GSL changes associated with: (i) ontogenic development and differentiation; (ii) oncogenic transformation and tumor progression. Various novel types of GSLs such as extended globo-series, sialyl Le(x) (SLe(x)), sialyl-dimeric-Le(x) (SLe(x)-Le(x)), dimeric-Le(x) (Le(x)-Le(x)), Le(y)-on-Le(x), dimeric-Le(a) (Le(a)-Le(a)), Le(b)-on-Le(a), etc. were identified as tumor-associated antigens. These studies provide an essential basis for up- or down-regulation of key glycosyltransferase genes controlling development, differentiation, and oncogenesis. GSL structures established in our laboratory are summarized in Table 1, and structural changes of GSLs associated with ontogenesis and oncogenesis are summarized in Sections 2 and 3. Based on these results, we endeavored to find out the cell biological significance of GSL changes, focused on (i) cell adhesion, e.g., the compaction process of preimplantation embryo in which Le(x)-to-Le(x), Gb4-to-GalGb4 or -nLc4 play major roles; and (ii) modulation of signal transduction through interaction of growth factor receptor tyrosine kinase with ganglioside, e.g., EGF receptor tyrosine kinase with GM3. Recent trends of studies on i and ii lead to the concept that GSL clusters (microdomains) are organized with various signal transducer molecules to form 'glycosignaling domains' (GSD). GSL-dependent adhesion occurs through clustered GSLs, and is coupled with activation of signal transducers (cSrc, Src family kinase, Rho A, etc.). Clustered GSLs involved in cell adhesion are recognized by GSLs on counterpart cells (carbohydrate-to-carbohydrate interaction), or by lectins (e.g., siglecs, selectins). Our major effort in utilization of GSLs in medical science has been for: (i) cancer diagnosis and treatment (vaccine development) based on tumor-associated GSLs and glycoepitopes; (ii) genetically defined phenotype for susceptibility to E. coli infection; (iii) clear identification of physiological E-selectin epitope (myeloglycan) expressed on neutrophils and myelocytes; (iv) characterization of sialyl poly-LacNAc epitopes recognized as male-specific antigens. Utilization of these GSLs or glycoepitopes in development of anti-adhesion approach to prevent tumor metastasis, infection, inflammation, or fertilization (i.e., contraceptive) is discussed. For each approach, development of mimetics of key GSLs or glycoepitopes is an important subject of future study. PMID- 11421355 TI - MUC1 in carcinoma-host interactions. AB - Many carcinoma-associated markers are glycoconjugates whose expression undergoes temporal or spatial regulation. Mucin-1 (MUC1), discovered through monoclonal antibody technology, is a well-documented example of such a molecule and influences numerous pathophysiological behaviors, such as the invasion and metastasis of carcinoma cells. Levels of MUC1 expression in carcinomas correlate with the clinical stage of the cancer and inversely correlate with the survival prospects of patients. The MUC1 immune response is known to provide a protective host defense mechanism against cancer. The multiple functions of MUC1 in carcinoma-host interactions are believed to be dependent on the polymorphic nature of MUC1, particularly its glycosylation status. PMID- 11421356 TI - Safe as mother's milk: carbohydrates as future anti-adhesion drugs for bacterial diseases. AB - The majority of infectious diseases are initiated by adhesion of pathogenic organisms to the tissues of the host. In many cases, this adhesion is mediated by lectins present on the surface of the infectious organism that bind to complementary carbohydrates on the surface of the host tissues. Lectin-deficient mutants often lack ability to initiate infection. Soluble carbohydrates recognized by the bacterial lectins block the adhesion of the bacteria to animal cells in vitro. Moreover, they have also been shown to protect against experimental infection by lectin-carrying bacteria in different organs of mammals such as mice, rabbits, calves and monkeys. In a phase II clinical trial, a pentasaccharide shown to have anti-adhesive activity against Streptococcus pneumoniae and Hemophilus influenzae in vitro failed to protect young children from nasopharyngeal colonization with these organisms and from developing otitis media. This could be because insufficient drug was delivered via nasal spray, because bacteria express multiple specificities, the inhibition of which may require a cocktail of oligosaccharides, or because children have different carbohydrate receptors from those of adults. The results of a clinical trial in which N-acetylneuraminyl(alpha2-3)lactose was administered orally to Helicobacter pylori positive patients in an effort to reduce or eradicate bacterial colonization, are awaited with interest. Although the high cost of production of the required oligosaccharides is falling with the recent introduction of enzymatic methods of synthesis, new technologies, in particular the use of engineered bacteria, promise to lower it even further. Attachment of the oligosaccharides to soluble polymeric carriers will increase greatly their effectiveness as antiadhesion agents. There is no doubt that anti-adhesive oligosaccharides will in the near future join the arsenal of drugs for the therapy of bacterial diseases. PMID- 11421357 TI - 3' splice site recognition in nematode trans-splicing involves enhancer-dependent recruitment of U2 snRNP. AB - Trans-splicing requires that 5' and 3' splice sites be independently recognized. Here, we have used mutational analyses and a sensitive nuclease protection assay to determine the mechanism of trans-3' splice site recognition in vitro. Efficient recognition of the 3' splice site is dependent upon both the sequence of the 3' splice site itself and enhancer elements located in the 3' exon. We show that the presence of three distinct classes of enhancers results in increased binding of U2 snRNP to the branchpoint region. Several lines of evidence strongly suggest that the increased binding of U2 snRNP is mediated by U2AF. These results expand the roles of enhancers in constitutive splicing and provide direct support for the recruitment model of enhancer function. PMID- 11421358 TI - Functional selection of splicing enhancers that stimulate trans-splicing in vitro. AB - The role of exonic sequences in naturally occurring trans-splicing has not been explored in detail. Here, we have identified trans-splicing enhancers through the use of an iterative selection scheme. Several classes of enhancer sequences were identified that led to dramatic increases in trans-splicing efficiency. Two sequence families were investigated in detail. These include motifs containing the element (G/C)GAC(G/C) and also 5' splice site-like sequences. Distinct elements were tested for their ability to function as splicing enhancers and in competition experiments. In addition, discrete trans-acting factors were identified. This work demonstrates that splicing enhancers are able to effect a large increase in trans-splicing efficiency and that the process of exon definition is able to positively enhance trans-splicing even though the reaction itself is independent of the need for the 5' end of U1 snRNA. Due to the presence of internal introns in messages that are trans-spliced, the natural arrangement of 5' splice sites downstream of trans-splicing acceptors may lead to a general promotion of this unusual reaction. PMID- 11421359 TI - The role of U2AF35 and U2AF65 in enhancer-dependent splicing. AB - Splicing enhancers are RNA sequence elements that promote the splicing of nearby introns. The mechanism by which these elements act is still unclear. Some experiments support a model in which serine-arginine (SR)-rich proteins function as splicing activators by binding to enhancers and recruiting the splicing factor U2AF to an adjacent weak 3' splice site. In this model, recruitment requires interactions between the SR proteins and the 35-kDa subunit of U2AF (U2AF35). However, more recent experiments have not supported the U2AF recruitment model. Here we provide additional evidence for the recruitment model. First, we confirm that base substitutions that convert weak 3' splice sites to a consensus sequence, and therefore increase U2AF binding, relieve the requirement for a splicing activator. Second, we confirm that splicing activators are required for the formation of early spliceosomal complexes on substrates containing weak 3' splice sites. Most importantly, we find that splicing activators promote the binding of both U2AF65 and U2AF35 to weak 3' splice sites under splicing conditions. Finally, we show that U2AF35 is required for maximum levels of activator-dependent splicing. We conclude that a critical function of splicing activators is to recruit U2AF to the weak 3' splice sites of enhancer-dependent introns, and that efficient enhancer-dependent splicing requires U2AF35. PMID- 11421360 TI - Differential alternative splicing activity of isoforms of polypyrimidine tract binding protein (PTB). AB - Polypyrimidine tract binding protein (PTB) is an RNA-binding protein that regulates splicing by repressing specific splicing events. It also has roles in 3'-end processing, internal initiation of translation, and RNA localization. PTB exists in three alternatively spliced isoforms, PTB1, PTB2, and PTB4, which differ by the insertion of 19 or 26 amino acids, respectively, between the second and third RNA recognition motif domains. Here we show that the PTB isoforms have distinct activities upon alpha-tropomyosin (TM) alternative splicing. PTB1 reduced the repression of TM exon 3 in transfected smooth muscle cells, whereas PTB4 enhanced TM exon 3 skipping in vivo and in vitro. PTB2 had an intermediate effect. The PTB4 > PTB2 > PTB1 repressive hierarchy was observed in all in vivo and in vitro assays with TM, but the isoforms were equally active in inducing skipping of alpha-actinin exons and showed the opposite hierarchy of activity when tested for activation of IRES-driven translation. These findings establish that the ratio of PTB isoforms could form part of a cellular code that in turn controls the splicing of various other pre-mRNAs. PMID- 11421361 TI - Substrate recognition by ADAR1 and ADAR2. AB - RNA editing catalyzed by ADAR1 and ADAR2 involves the site-specific conversion of adenosine to inosine within imperfectly duplexed RNA. ADAR1- and ADAR2-mediated editing occurs within transcripts of glutamate receptors (GluR) in the brain and in hepatitis delta virus (HDV) RNA in the liver. Although the Q/R site within the GluR-B premessage is edited more efficiently by ADAR2 than it is by ADAR1, the converse is true for the +60 site within this same transcript. ADAR1 and ADAR2 are homologs having two common functional regions, an N-terminal double-stranded RNA-binding domain and a C-terminal deaminase domain. It is neither understood why only certain adenosines within a substrate molecule serve as targets for ADARs, nor is it known which domain of an ADAR confers its specificity for particular editing sites. To assess the importance of several aspects of RNA sequence and structure on editing, we evaluated 20 different mutated substrates, derived from four editing sites, for their ability to be edited by either ADAR1 or ADAR2. We found that when these derivatives contained an A:C mismatch at the editing site, editing by both ADARs was enhanced compared to when A:A or A:G mismatches or A:U base pairs occurred at the same site. Hence substrate recognition and/or catalysis by ADARs could involve the base that opposes the edited adenosine. In addition, by using protein chimeras in which the deaminase domains were exchanged between ADAR1 and ADAR2, we found that this domain played a dominant role in defining the substrate specificity of the resulting enzyme. PMID- 11421362 TI - A purine-rich intronic element enhances alternative splicing of thyroid hormone receptor mRNA. AB - The mammalian thyroid hormone receptor gene c-erbAalpha gives rise to two mRNAs that code for distinct isoforms, TRalpha1 and TRalpha2, with antagonistic functions. Alternative processing of these mRNAs involves the mutually exclusive use of a TRalpha1-specific polyadenylation site or TRalpha2-specific 5' splice site. A previous investigation of TRalpha minigene expression defined a critical role for the TRalpha2 5' splice site in directing alternative processing. Mutational analysis reported here shows that purine residues within a highly conserved intronic element, SEa2, enhance splicing of TRalpha2 in vitro as well as in vivo. Although SEalpha2 is located within the intron of TRalpha2 mRNA, it activates splicing of a heterologous dsx pre-mRNA when located in the downstream exon. Competition with wild-type and mutant RNAs indicates that SEalpha2 functions by binding trans-acting factors in HeLa nuclear extract. Protein-RNA crosslinking identifies several proteins, including SF2/ASF and hnRNP H, that bind specifically to SEalpha2. SEalpha2 also includes an element resembling a 5' splice site consensus sequence that is critical for splicing enhancer activity. Mutations within this pseudo-5' splice site sequence have a dramatic effect on splicing and protein binding. Thus SEa2 and its associated factors are required for splicing of TRalpha2 pre-mRNA. PMID- 11421363 TI - Imaging of single hairpin ribozymes in solution by atomic force microscopy. AB - The hairpin ribozyme is a short endonucleolytic RNA motif isolated from a family of related plant virus satellite RNAs. It consists of two independently folding domains, each comprising two Watson-Crick helices flanking a conserved internal loop. The domains need to physically interact (dock) for catalysis of site specific cleavage and ligation reactions. Using tapping-mode atomic force microscopy in aqueous buffer solution, we were able to produce high quality images of individual hairpin ribozyme molecules with extended terminal helices. Three RNA constructs with either the essential cleavage site guanosine or a detrimental adenosine substitution and with or without a 6-nt insertion to confer flexibility to the interdomain hinge show structural differences that correlate with their ability to form the active docked conformation. The observed contour lengths and shapes are consistent with previous bulk-solution measurements of the transient electric dichroism decays for the same RNA constructs. The active docked construct appears as an asymmetrically docked conformation that might be an indication of a more complicated docking event than a simple collapse around the interdomain hinge. PMID- 11421364 TI - Identification of cis-acting elements involved in 3'-end formation of Saccharomyces cerevisiae 18S rRNA. AB - In yeast, the 3' end of mature 18S rRNA is generated by endonucleolytic cleavage of the 20S precursor at site D. Available data indicate that the major cis-acting elements required for this processing step are located in relatively close proximity to the cleavage site. To identify these elements, we have studied the effect of mutations in the mature 18S and ITS1 sequences neighboring site D on pre-rRNA processing in vivo. Using clustered point mutations, we found that alterations in the sequence spanning site D from position -5 in 18S rRNA to +6 in ITS1 reduced the efficiency of processing at this site to different extents as demonstrated by the lower level of the mature 18S rRNA and the increase in 20S pre-rRNA in cells expressing only mutant rDNA units. More detailed analysis revealed an important role for the residue located 2 nt upstream from site D (position -2), whereas sequence changes at position -1, +1, and +2 relative to site D had no effect. The data further demonstrate that the proposed base pairing between the 3' end of 18S rRNA and the 5' end of ITS1 is not important for efficient and accurate processing at site D, nor for the formation of functional 40S ribosomal subunits. These results were confirmed by analyzing the accumulation of the D-A2 fragment derived from the mutant 20S pre-rRNA in cells that lack the Xrn1p exonuclease responsible for its degradation. The latter results also showed that the accuracy of cleavage was affected by altering the spacer sequence directly downstream of site D but not by mutations in the 18S rRNA sequence preceding this site. PMID- 11421365 TI - An unexpected, conserved element of the U3 snoRNA is required for Mpp10p association. AB - The U3 small nucleolar ribonucleoprotein (snoRNP) is composed of a small nucleolar RNA (snoRNA) and at least 10 proteins. The U3 snoRNA base pairs with the pre-rRNA to carry out the A0, A1, and A2 processing reactions that lead to the release of the 18S rRNA from the nascent pre-rRNA transcript. The yeast U3 snoRNA can be divided into a short 5' domain (nt 1-39) and a larger 3' domain (73 to the 3' end) separated by a stretch of nucleotides called the hinge region (nt 40-72). The sequences required for pre-rRNA base pairing are found in the 5' domain and hinge region whereas the 3' domain is largely covered with proteins. Mpp10p, one of the protein components unique to the U3 snoRNP, plays a role in processing at the A1 and A2 sites. Because of its critical role in U3 snoRNP function, we determined which sequences in the U3 snoRNA are required for Mpp10p association. Unlike fibrillarin and all the previous U3 snoRNP components studied in this manner, sequences in the 3' domain are not sufficient for Mpp10p association. Instead, a conserved sequence element in the U3 snoRNA hinge region is required, placing Mpp10p near the 5' domain that carries out the pre-rRNA base pairing interactions in the functional center of the U3 snoRNP. PMID- 11421366 TI - The 3'-end-processing factor CPSF is required for the splicing of single-intron pre-mRNAs in vivo. AB - We describe a new approach to elucidate the role of 3'-end processing in pre-mRNA splicing in vivo using the influenza virus NS1A protein. The effector domain of the NS1A protein, which inhibits the function of the CPSF and PABII factors of the cellular 3'-end-processing machinery, is sufficient for the inhibition of not only 3'-end formation but also the splicing of single-intron pre-mRNAs in vivo. We demonstrate that inhibition of the splicing of single-intron pre-mRNAs results from inhibition of 3'-end processing, thereby establishing that 3'-end processing is required for the splicing of a 3' terminal intron in vivo. Because the NS1A protein causes a global suppression of 3'-end processing in trans, we avoid the ambiguities caused by the activation of cryptic poly(A) sites that occurs when mutations are introduced into the AAUAAA sequence in the pre-mRNA. In addition, this strategy enabled us to establish that the function of a particular 3'-end processing factor, namely CPSF, is required for the splicing of single-intron pre mRNAs in vivo: splicing is inhibited only when the effector domain of the NS1A protein binds and inhibits the function of the 30-kDa CPSF protein in 3'-end formation. In contrast, the 3'-end processing factor PABII is not required for splicing. We discuss the implications of these results for cellular and influenza viral mRNA splicing. PMID- 11421367 TI - Molecular epidemiology of malaria in Yaounde, Cameroon. VII. Analysis of recrudescence and reinfection in patients with uncomplicated falciparum malaria. AB - In an endemic area where malaria transmission is intense and continuous, reappearance of asexual parasites may be ascribed to either recrudescence or reinfection. To distinguish between recrudescence and reinfection after oral treatment with chloroquine, amodiaquine, pyronaridine, sulfadoxine-pyrimethamine, halofantrine, or artesunate, three polymorphic markers (circumsporozoite protein, merozoite surface antigens 1 and 2) from pre-treatment and post-treatment samples were amplified by the polymerase chain reaction, and the in vitro response to chloroquine was determined for comparison. Of 52 paired samples, 22 (42%) were reinfections. Recrudescence occurred more frequently on or before Day 14 (22 of 30 cases, 73%). Except for one case, all reinfections were observed beyond Day 14. The phenotype determination was not sufficiently precise to distinguish between recrudescence and reinfection. Our results suggest that beyond Day 14 (and until Day 42), recrudescence and reinfection cannot be distinguished at our study site unless molecular techniques are used and that some results derived from the polymerase chain reaction need to be compared with the microscopic examination of thick blood smear to exclude gametocyte carriers without asexual parasites after treatment. PMID- 11421368 TI - The epidemiology of Plasmodium falciparum malaria in two Cameroonian villages: Simbok and Etoa. AB - In support of ongoing immunologic studies on immunity to Plasmodium falciparum, demographic, entomologic, parasitologic, and clinical studies were conducted in two Cameroonian villages located 3 km apart. Simbok (population = 907) has pools of water present year round that provide breeding sites for Anopheles gambiae, whereas Etoa (population = 485) has swampy areas that dry up annually in which A. funestus breed. Results showed that individuals in Simbok receive an estimated 1.9 and 1.2 infectious bites per night in the wet and dry season, respectively, whereas individuals in Etoa receive 2.4 and 0.4 infectious bites per night, respectively. Although transmission patterns differ, the rate of acquisition of immunity to malaria appears to be similar in both villages. A prevalence of 50 75% was found in children < 10 years old, variable levels in children 11-15 years old, and 31% in adults. Thus, as reported in other parts of Africa, individuals exposed to continuous transmission of P. falciparum slowly acquired significant, but not complete, immunity. PMID- 11421369 TI - The contrasting epidemiology of Cyclospora and Cryptosporidium among outpatients in Guatemala. AB - We compared epidemiologic characteristics of Cryptosporidium and Cyclospora in surveillance data from outpatient departments in Guatemala. Routinely-submitted stool specimens were screened by microscopy. Age, sex, and symptom data were collected. Cyclospora was detected in 117 (2.1%) and Cryptosporidium in 67 (1.2%) of 5,520 specimens. The prevalence of Cyclospora peaked in the warmer months, while Cryptosporidium was most common in the rainy season. Both affected children more than adults, but Cryptosporidium affected children at a younger age than Cyclospora (median age 2 years versus 5 years; P < 0.001). Cyclospora showed a stronger association with diarrhea than Cryptosporidium, even when data were stratified by age. These contrasts may reflect differences in the relative importance of transmission modes, the frequency of exposure, and the development of immunity. PMID- 11421370 TI - Hepatitis C in a community in Upper Egypt: I. Cross-sectional survey. AB - The prevalence of antibody to hepatitis C virus (anti-HCV) was determined in a cross-sectional survey in a village in Upper Egypt. Exposure and demographic characteristics were obtained through a questionnaire. Antibody to hepatitis C virus was assessed using a second generation enzyme immunoassay, and the presence of HCV RNA was tested using a reverse transcriptase-polymerase chain reaction. Collection of blood samples was targeted at those > or = 5 years old, and obtained from 62.8%. This report describes the community, the HCV infection characteristics of the subjects, and evaluates some factors associated with presence of anti-HCV. Of the 6,031 participants, 522 (8.7%) were anti-HCV positive. Prevalence was higher among males than females (11.3% versus 6.5%; P < 0.001). It was greater among those > 30 years of age than among those < or = 30 years of age (20.0% versus 3.6%; P < 0.001). Those who were less educated, farmed, provided health care, and were currently married had a significantly higher anti-HCV prevalence than those who were not; however, these associations were not significant after adjusting for age. Although active infections with Schistosoma haematobium were not associated with anti-HCV, a history of past infection was (age-adjusted risk ratio [RR] = 2.1, 95% confidence interval [CI] = 1.8, 2.4); 134 persons who had a history of receiving parenteral anti schistosomal therapy had a higher age-adjusted RR (3.0; 95% CI = 2.5, 3.7) for anti-HCV than those who did not. Hepatitis C virus RNA was detected in 62.8% of the anti-HCV positive subjects, without significant variation by age, gender, education, or marital status. The prevalence of anti-HCV in Upper Egypt is high, albeit lower than in Lower Egypt, with continuing but limited transmission indicated by the lower prevalence in residents < or = 30 years old. PMID- 11421371 TI - Hepatitis C in Peru: risk factors for infection, potential iatrogenic transmission, and genotype distribution. AB - A large seroepidemiologic and genotyping study of hepatitis C virus (HCV) was conducted in Lima, Peru, during the periods of 1986 to 1993 (cohort A) and 1994 (cohort B). Anti-HCV seroprevalence rates were 15.6% (216 of 1,389) and 11.7% (168 of 1,438), respectively. Low rates were seen among volunteer blood donors (1.1% and 0.8%). Anti-HCV rates were much higher among patients undergoing hemodialysis (43.7% and 59.3%), hemophiliacs (60.0% and 83.3%), in those more than 39 years old (18.2% and 26.0%), in females (25.0% and 27.4%), and in less educated persons (16.9%). Age- and gender-adjusted risk factors in cohort B included blood transfusion history (adjusted odds ratio [AOR] = 29.8), prior organ transplantation (AOR = 9.1) or a history of hepatitis (AOR = 4.9), previous hospitalization (AOR = 3.7), a history of intravenous drug use (AOR = 3.5), prior major surgery (AOR = 2.6), a history of acupuncture (AOR = 2.1), previous dental procedures (AOR = 1.2), and prior medical injections (AOR = 1.04). The most prevalent HCV genotype was type 1 (86%), followed by type 3 (10%) and type 2 (2%). Transmission through unsafe injection-related and medical/dental procedures appears to play an important role in HCV infection among Peruvians. PMID- 11421372 TI - Asymptomatic infection and risk factors for leptospirosis in Nicaragua. AB - As part of an investigation of a 1995 outbreak of leptospirosis in Nicaragua, a cross-sectional serologic survey was conducted in the town of El Sauce. Of 566 persons, 85 (15%) were positive for IgM anti-Leptospira antibodies, indicating recent leptospirosis infection. Asymptomatic leptospirosis infection was common, with only 25 (29.4%) of the 85 seropositive inhabitants reporting a febrile illness in the 2 months before the survey. Multivariable analysis revealed that having an indoor water source remained independently protective against leptospirosis. Gathering wood was independently associated with infection. These findings suggest that asymptomatic infection with Leptospira is common in endemic areas of Leptospira transmission. Improvement in water sanitation and behavioral modifications to reduce environmental exposure may reduce the risk of leptospirosis in endemic regions. PMID- 11421373 TI - Detection of pathogenic Leptospira spp. infections among mammals captured in the Peruvian Amazon basin region. AB - To identify potential zoonotic reservoirs of pathogenic leptospires in the Peruvian Amazon basin, wild mammals were trapped from July 1997 to December 1998 near the city of Iquitos. After extraction of nucleic acids from animal kidneys, DNA of pathogenic leptospires was identified by polymerase chain reaction (PCR) assays using one of two primer sets, one amplifying a region of the 23S rRNA gene, and the other amplifying a gene fragment specific for Leptospira spp (G1/G2 primers). Overall, 29% (40 of 136) of the mammals tested showed evidence of renal infection by Leptospira spp., including 20% (13 of 64) of the rodents, 39% (20 of 51) of the marsupials, and 35% (7 of 20) of the chiropterans (bats). Marsupials and chiropterans were implicated as more significant reservoir hosts of leptospires pathogenic to humans than previously recognized. PMID- 11421374 TI - Antimicrobial agent resistance in bacterial isolates from patients with diarrhea and urinary tract infection in the Sudan. AB - Antimicrobial sensitivity tests were performed on four-hundred and ninety-seven bacterial isolates from Sudanese patients with diarrhea or urinary tract infections. Shigella dysenteriae type I and enteropathogenic Escherichia coli showed high resistance rates (percentage of isolates showing antibiotic resistance) against the commonly-used antimicrobial agents: ampicillin, amoxicillin, chloramphenicol, tetracycline, cotrimoxazole, nalidixic acid, sulfonamide, and neomycin, and were completely sensitive to ciprofloxacin. Eighteen resistance patterns against nine antimicrobial agents tested were observed in enteric pathogens. Resistance to ampicillin, amoxicillin, tetracycline, cotrimoxazole, and sulfonamide was the most frequent pattern. The common urinary pathogens, E. coli, Klebsiella pneumoniae, and Proteus mirabilis showed high rates of resistance to ampicillin, amoxicillin, cotrimoxazole, tetracycline, sulfonamide, trimethoprim, streptomycin, and carbenicillin. We recommend that physicians seek updated knowledge of the common antibiotic sensitivity patterns when starting empirical antibiotic therapy in Sudanese patients with diarrhea or urinary tract infection. PMID- 11421375 TI - The efficacy and tolerability of triclabendazole in Cuban patients with latent and chronic Fasciola hepatica infection. AB - Current chemotherapy for the treatment of infections caused by the liver fluke Fasciola hepatica is not satisfactory. Therefore, the efficacy and tolerability of triclabendazole (TCZ) was assessed for this indication. Eighty-two patients (51 female, 31 male, age 15-81 yr, mean 42 yr) with chronic or latent F. hepatica infection refractory to previous anti-helminthic chemotherapy were enrolled in a 60-day open, non-comparative trial. Patients received 20 mg/kg TCZ as two doses of 10 mg/kg administered after food 12 hr apart. Efficacy of treatment was assessed by stool microscopy, determination of Fasciola excretory-secretory antigen (FES) in feces, and by ultrasonography (US) which were systematically performed pre-therapy and on Days 1-7, 15, 30, and 60 post-therapy. For continuous safety assessment, patients were hospitalized during the first week after therapy and then monitored at home for the appearance of any adverse events. Clinical chemistry and hematology tests were carried out on Days 1, 3, 7, 15, and 60, and whenever an adverse effect occurred possibly related to therapy. Seventy-one (92.2%) of the 77 patients who completed the 60-day follow-up period became egg-negative. Efficacy of therapy was supported by the disappearance or decrease of FES antigen and of ultrasonography abnormalities. In the 6 remaining patients, parasitological cure was achieved by another single TCZ dose of 10 mg/kg on Day 60. A total of 74 adverse events possibly related to therapy was reported by 54 patients. The most important adverse event was colic-like abdominal pain (40 patients [49%]) consistent with the expulsion of the parasite through the bile ducts as confirmed by US on Days 2-7. Most adverse events (53) were graded as mild, 20 as moderate, and only 1 as severe (a biliary colic responding to spasmolytic therapy within two hours). Triclabendazole 20 mg/kg is an effective therapy for the treatment of F. hepatica infection in patients who have failed to respond to other antihelminthic agents. Biliary colics reflecting the expulsion of dead or damaged parasites usually occur during Day 3-7 and respond well to spasmolytic therapy. PMID- 11421376 TI - Effect of dose increase or cimetidine co-administration on albendazole bioavailability. AB - The low bioavailability of albendazole affects the therapeutic response in patients with echinococcosis. Cimetidine co-administration is reported to improve bioavailability. To analyze the assumed dose-dependent bioavailability of albendazole, we administered 5 to 30 mg/kg albendazole to 6 male volunteers in a randomized cross-over study. To assess the effect of cimetidine (10 mg/kg twice daily), the drug was given with albendazole (20 mg/kg). A dose-dependent bioavailability was not observed. This was due to inter-individual variability of the maximal concentration (Cmax 38%-72%) of albendazole sulphoxide (ABZSX), the active metabolite of albendazole. Cmax was 0.21+/-0.14 mg/L after 5 mg/kg and 0.39+/-0.19 mg/L after 30 mg/kg albendazole (P = 0.217). Cimetidine tended to decrease Cmax by 52% (P = 0.109) and significantly inhibited ABZSX breakdown as indicated by the prolongation of ABZSX elimination half-life from 7.4+/-3.3 hr to 19.0+/-11.7 hr (P = 0.028). Remarkably, the inter-individual variability of Cmax was significantly lower during cimetidine co-administration: 14% versus 72%. PMID- 11421377 TI - Atovaquone plus proguanil versus halofantrine for the treatment of imported acute uncomplicated Plasmodium falciparum malaria in non-immune adults: a randomized comparative trial. AB - In endemic zones, the atovaquone-proguanil (AP) combination is well tolerated and effective in treating acute, uncomplicated malaria. Trials involving non-immune patients are lacking, however. We conducted a randomized, multicenter open-label trial to determine the efficacy and tolerability of the AP combination (1,000 mg + 400 mg once daily for 3 days) in comparison with halofantrine (HF) (1,500 mg in 3 doses) in non-immune adults with imported uncomplicated Plasmodium falciparum malaria. Follow-up visits were programmed on Days 7, 14, 21, 28, and 35 after hospital discharge. Out of 48 patients enrolled in the study, 41 were assessable for the cure rate (21 in the AP group and 20 in the HF group). All the patients were cured. The mean parasite clearance time was longer (63+/-23 hours) in the AP group than in the HF group (48+/-15 hours) (P = 0.02). The frequency of gastrointestinal adverse events was higher in the AP group. No noteworthy electrocardiographic changes were observed, particularly in the QTc interval. The AP combination appears to be a valuable alternative treatment in non-immune adults. PMID- 11421378 TI - 16alpha-bromoepiandrosterone, a dehydroepiandrosterone (DHEA) analogue, inhibits Plasmodium falciparum and Plasmodium berghei growth. AB - Dehydroepiandrosterone (DHEA) and its analogue, 16alpha-bromoepiandrosterone (alpha-epi-Br), may have activity against viral and parasitic infections, including human immunodeficiency virus (HIV) and Cryptosporidium parvum. Therefore, we evaluated its antimalarial effects on Plasmodium falciparum and Plasmodium berghei. In vitro, chloroquine (CQ)-sensitive and resistant strains of P. falciparum parasitized red blood cells were incubated with escalating doses of alpha-epi-Br or CQ. In vivo, 62 rats were infected with P. berghei and treated with CQ or alpha-epi-Br. At the highest doses tested against a CQ-sensitive strain, parasitemias decreased from 25.4% in the saline control group to 4.3% and 4.8% in the alpha-epi-Br and CQ groups, respectively (P < 0.05). Against two CQ resistant strains, parasitemias decreased from 22.3-28.8% and 24.8-30% in the CQ and saline groups, respectively, to 2.5-2.7% in the alpha-epi-Br groups (P = 0.003). In vivo, on Day 4, parasitemias decreased from 23% in the saline group to 9-12% and 12% in the in alpha-epi-Br and CQ groups, respectively (P < 0.05). These data demonstrate that alpha-epi-Br shows activity against CQ-sensitive and resistant strains of P. falciparum in vitro. At the doses tested against P. berghei in vivo in rats, alpha-epi-Br is comparable to CQ. PMID- 11421379 TI - Use of intravaginal microbicides to prevent acquisition of Trichomonas vaginalis infection in Lactobacillus-pretreated, estrogenized young mice. AB - D2A21, a novel peptide antibiotic has in vitro activity against a wide spectrum of sexually transmitted diseases (STD). In this study we tested the hypothesis that intravaginal D2A21 would interfere with acquisition of Trichomonas vaginalis infection in a modified mouse model. T. vaginalis infections of estrogenized young mice pretreated with Lactobacillus vaginalis or Lactobacillus rhamnosus were more frequent and persistent than those in mice pre-treated with Lactobacillus gasseri or Lactobacillus acidophilus. One hundred percent T. vaginalis infection was achieved for 2-4 days post-challenge when intravaginal L. rhamnosus pre-treatments were given to estrogenized mice 48 hr prior to a single T. vaginalis challenge. Estrogenized mice pre-treated with L. rhamnosus were pre medicated with intravaginal placebo gel, 0.5% or 2% D2A21 gel, or 500 microg/mL metronidazole gel prior to T. vaginalis challenge. Both 2% D2A21 and metronidazole gels were significantly more efficacious (10% or none infected) than placebo gel (53% infected) in preventing vaginal T. vaginalis infections in mice. PMID- 11421380 TI - Clinical trial of sequential treatments of moderately severe and severe malaria with dihydroartemisinin suppository followed by mefloquine in Thailand. AB - One hundred and fifty patients with severe falciparum malaria were administered sequential combination of dihydroartemisinin suppository followed by an oral mefloquine tablet. Dihydroartemisinin suppositories (80 mg/capsule) were given rectally once daily for 3 days with a total dose of 8-10 mg/kg. Two doses of mefloquine, 15 mg/kg/dose and 10 mg/kg/dose, were given at 72 hr and 84 hr, respectively. The mean [SD] parasite clearance time and fever clearance time were 46.1 [15.7] hr and 82.5 [59.6] hr, respectively. No death or major adverse drug effects occurred. The cure rate at 28 days of inpatient follow-up was 95% (113 of 119 patients). In severe malaria, dihydroartemisinin suppository followed by oral mefloquine is a suitable alternative treatment to parenteral drugs. PMID- 11421381 TI - Self-testing for falciparum malaria with antigen-capture cards by travelers with symptoms of malaria. AB - Kits for self-diagnosis of malaria by travelers in remote areas have been advocated. Antigen-capture test cards work reliably in the laboratory, but there is limited data regarding self-testing by ill patients. One hundred and fifty three symptomatic patients presenting for a malaria test attempted self-diagnosis using these kits. Nine percent failed to perform a valid test. Against blood-film proven malaria, specificity was 97% (95% confidence intervals [CI] 93-99%) and sensitivity 95% (95% CI 74-99%). Although these results are encouraging, technical problems need to be addressed and test sensitivity proven by additional field-testing before this technology can be marketed to travelers. PMID- 11421382 TI - Safety and immunogenicity of a new equine tetanus immunoglobulin associated with tetanus-diphtheria vaccine. AB - In a single-center double-blind, randomized trial in West Africa, we evaluated the safety and immunogenicity of a new pasteurized, pepsin-digested equine tetanus immunoglobulin (heat-treated equine tetanus immunoglobulin [HT-ETIG]) in the post-exposure prophylaxis of tetanus compared with the reference product, equine tetanus immunoglobulin (ETIG). A total of 134 adults presenting to Garoua Hospital, Cameroon with a tetanus-prone wound were randomized to receive a 3,000 international units (IU) intramuscular injection (deltoid) of either HT-ETIG or ETIG, simultaneously with a tetanus-diphtheria vaccine. No serious adverse reactions were reported. The incidences of local and systemic reactions were similar in the two groups. Repeated measures of equine tetanus-antibody levels measured from Day 0 to Day 28 showed that titers were significantly higher in the HT-ETIG group (P = 0.017). At Day 7, a higher percentage of subjects in the HT ETIG group had equine antibody levels > or = 0.1 IU/ml (80.4% versus 37.9%; P < 0.0001). No cases of tetanus occurred during the follow-up, attesting to the efficacy of the combined prophylactic treatment. PMID- 11421383 TI - Tc45, a dimorphic Trypanosoma cruzi immunogen with variable chromosomal localization, is calreticulin. AB - We demonstrate that Tc45, a polypeptide described as an immunogenetically restricted Trypanosoma cruzi antigen in mice, is calreticulin, a dimorphic molecule encoded by genes with variable chromosomal distribution. Previously we showed that IgG from A.SW (H2s) mice immunized with T. cruzi trypomastigotes or epimastigotes and sera from infected humans recognize Tc45, a 45 kD parasite polypeptide. Herein we describe the cloning, sequencing, and expression of the Tc45 gene. A 98% homology in the deduced amino acid sequence was found with a T. cruzi calreticulin-like molecule and 41% with Leishmania donovani and human calreticulin. In the T. cruzi CL Brener clone and in the Tulahuen strain, the gene is located in two and four chromosomes, respectively. Calreticulin was detected in several T. cruzi clones, in the Tulahuen strain, and in T. rangeli, displaying alternative 43 and 46 kD forms. PMID- 11421384 TI - Inhibition of local hemorrhage and dermonecrosis induced by Bothrops asper snake venom: effectiveness of early in situ administration of the peptidomimetic metalloproteinase inhibitor batimastat and the chelating agent CaNa2EDTA. AB - The effectiveness of the chelating agent CaNa2EDTA and the peptidomimetic matrix metalloproteinase inhibitor batimastat (BB-94) to inhibit local tissue damage induced by Bothrops asper snake venom was studied in mice. Both compounds totally inhibited proteolytic, hemorrhagic, and dermonecrotic effects, and partially reduced edema-forming activity, when incubated with venom prior to injection. Much lower concentrations of batimastat than of CaNa2EDTA were required to inhibit these effects. In addition, batimastat, but not CaNa2EDTA, partially reduced myotoxic activity of the venom. When the inhibitors were administered at various time intervals after envenomation at the same site of venom injection, both compounds were effective in neutralizing local hemorrhage and dermonecrosis if administered rapidly after venom. Inhibition was not as effective as the time lapse between venom and inhibitor injections increased. Owing to the relevance of metalloproteinases in the pathogenesis of local tissue damage induced by B. asper and other pit viper venoms, it is suggested that administration of peptidomimetic metalloproteinase inhibitors or CaNa2EDTA at the site of venom injection may represent a useful alternative to complement antivenoms in the neutralization of venom-induced local tissue damage. PMID- 11421385 TI - An aid to accurate planning of alar reconstruction in rhinoplasty. AB - Accurate placement of intranasal incisions to achieve external alar symmetry is difficult. A detailed anatomical knowledge of intranasal anatomy is required to allow an appropriate incision to be made within the nasal vestibule. By marking the level of the intracartilaginous incision on the external surface using a pair of alice forceps, the marking is subsequently transferred onto the vestibular nasal surface. The initial incision can therefore be made on the vestibular nasal surface along the marking made by the alice forceps. PMID- 11421386 TI - Selection for rehabilitation: acute care discharge patterns for stroke and orthopaedic patients. AB - This study aimed to develop predictive models to assist clinicians working in acute care to determine which stroke and lower limb orthopaedic patients should be discharged to nursing homes, inpatient rehabilitation, or home. The subjects were 223 patients with stroke and 125 patients with lower limb orthopaedic problems, aged 60 years and over, and the 22 acute care and rehabilitation clinicians who cared for these patients. Patient status was measured within 72 hours of discharge using the Adult FIM(SM) (Guide for the Uniform Data Set for Medical Rehabilitation, 1993) and ten additional items from either the RICFAS (Rehabilitation Institute of Chicago, 1987) or developed for this research. Four mathematical models were developed to predict discharge destination using stepwise discriminant function analysis. Using between three and seven items from the FIM(SM) alone or FIM(SM) plus additional variables, these models correctly classified between 74.9% and 80.5% of patients. In conclusion, clinicians are offered a selection of models to guide their discharge destination decisions for two large groups of patients. While these models cannot replace clinical judgement, they increase the transparency of decisions. The use of objective models to guide clinical decisions are essential, particularly given increasing pressure to justify patient access to costly rehabilitation services. PMID- 11421387 TI - The rehabilitation process of children with motor disabilities in the Dutch therapeutic toddler class: main phases and parent involvement. AB - This study describes the implementation of the main phases of the rehabilitation process of children with motor disabilities who visit Dutch therapeutic toddler classes. Parent involvement, as well as parents' and professionals' satisfaction were also investigated. It was found that in about half of the cases there was no initial assessment of the child, nor a jointly formulated intervention plan. When there was an assessment, several quality standards were not met. In most cases parents were involved in the evaluation phase of the rehabilitation process. However, the proportion of parents involved in the phases of assessment and intervention planning were much lower. The results are discussed within the context of current developments in The Netherlands with respect to team communication and recent research findings. PMID- 11421388 TI - Functional independence measure (FIM) assessing outcome in medical rehabilitation of neurologically ill adolescents. AB - With the Functional Independence measure, which is a reliable and valid instrument, data can be collected for large groups of persons with different disabilities. For the first time the FIM has been used for outcome assessment in medical rehabilitation of a cohort of neurologically ill adolescents. The instrument is appropriate for this purpose. The severity of disability, which changes during rehabilitation of adolescents, is adequately estimated with FIM. The motor FIM and the cognitive FIM can be summed up separately for evaluation. In our investigation the construct validity of the cognitive FIM was demonstrated: the cognitive FIM showed a high correlation with the degree of neuropsychological impairment following brain damage. Psychiatric disorder does not correlate with the motor FIM and the severity of disability. PMID- 11421389 TI - Impairment, disability, handicap and participation in post-poliomyelitis subjects. PMID- 11421390 TI - The establishment of a community multiple sclerosis team. PMID- 11421391 TI - Student teachers' willingness to mainstream pupils with special needs in relation to track and severity of the disability. PMID- 11421392 TI - The effect of congenital handicap on personality development in adolescence. PMID- 11421393 TI - Validation of the Complete Minnesota Dexterity Test for adults with moderate graded mental retardation in Hong Kong. PMID- 11421394 TI - Coping responses in patients with spinal cord injury and adjustment difficulties. PMID- 11421395 TI - Prosthetic rehabilitation following below-knee amputation in elderly persons. PMID- 11421396 TI - Relating familial stress to the psychosocial adjustment of adolescents with Duchenne muscular dystrophy. AB - Adolescence is a time that offers significant challenges to families dealing with Duchenne muscular dystrophy (DMD). A major developmental task in the adjustment process for the adolescent is the achievement of a sense of self-identity. Mastering stress in families of adolescents with DMD has also been shown to be a major task in the adjustment process for the family. In this study, the relationships between familial stress and psychosocial adjustment in adolescents with DMD were investigated. The study participants (36 caregivers and 32 adolescents) were all registered with the Muscular Dystrophy Association of Canada and all resided in the province of Ontario. The Offer Self-Image Questionnaire for Adolescents (Offer et al., 1982, 1989) was used to measure psychosocial adjustment of adolescents with DMD. Family stress was measured by the Questionnaire on Resources and Stress (Holroyd, 1987). All but a few of the participants were interviewed and completed self-report instruments in their own homes. The other families were interviewed in other settings (e.g., clinic, adolescent's school). The results indicate that, overall and in several specific areas, the adolescents demonstrate lower levels of psychosocial adjustment than their normal reference group. The results also indicate that familial stress is not related to sociodemographic variables, but is associated with psychosocial adjustment in the adolescent (P = 0.0004), and with intellectual function of the adolescent (P = 0.001). In fact, the results of a regression analysis show that the level of stress experienced by the family predicts the degree of psychosocial adjustment of the adolescent with DMD (R2 = 0.38, P = 0.0003). These findings underscore the importance of a holistic approach with these families. Limitations of the study, and implications of the results, are discussed in terms of clinical practice and future research. PMID- 11421397 TI - Current use of outcome measures for stroke and low back pain rehabilitation in five European countries: first results of the ACROSS project. AB - Objectives were to obtain a view on the current use of outcome measures for stroke and low back pain rehabilitation in five European countries. A postal questionnaire, comparable for different cultural situations, was distributed in August 1998 to 581 rehabilitation facilities in Ireland, Germany, Italy, Austria and the Netherlands. Of these, 102 settings responded. In stroke rehabilitation the Barthel Index is the dominant outcome measure, followed by the Functional Independence Measure and the Frenchay Activities Index. Besides the Visual Analogue Scale, the most used outcome measures in low back pain rehabilitation were the Oswestry Pain Disability Questionnaire and the Roland Disability Questionnaire. Outcome measures are more frequently used in stroke than in low back pain rehabilitation. The purpose of use is mainly for the measurement of effectiveness, while a relation with quality management is seldom made. There appears to be little agreement on which outcome measures to use. Little attention was found to have been given to the assessment of handicap, quality of life and patient satisfaction. PMID- 11421398 TI - A perception of the advantages of a personal dental service. PMID- 11421399 TI - Influence of dental care systems on dental status. A comparison between two countries with different systems but similar living standards. AB - OBJECTIVE: To evaluate the influence of two different dental care systems on dental status, taking into account relevant socio-economic factors. BASIC RESEARCH DESIGN: Questionnaire studies on randomly sampled subjects in Denmark and Sweden using questionnaire forms as identical as possible with regard to the different languages. SETTING: The studies were performed late in 1998 in both countries. PARTICIPANTS: Questionnaires were sent to 1,175 subjects aged 45-69 years in Denmark (response rate 73%) and to 1,001 subjects aged 55-79 years in Sweden (response rate 67%). MAIN OUTCOME MEASURES: Questions about dental status and about socioeconomic factors and attitudes toward dental care were included. In logistic regression models, various dichotomies of dental conditions were used as dependent variables. State (Denmark vs. Sweden) was used as an independent variable together with socioeconomic factors and attitudes. RESULTS: There were great differences between the countries in dental status. In the regression model with 'wearing removable denture(s)' as the dependent variable, state was the strongest predictor with an OR of above 4.1 for Denmark compared to Sweden. much stronger than variables such as age, income, education and residence. CONCLUSION: The results indicate that the Swedish dental care system has been superior to the Danish one regarding dental status in middle aged and older populations in these two countries. PMID- 11421400 TI - Self-reported dental health, treatment need and attendance among older adults in two areas of Finland. AB - OBJECTIVE: The purpose of the study was to assess differences in dental health, need for care and attendance among older adults in two areas of Finland with contrasting socio-cultural and economic backgrounds. A further aim was to analyse and compare the determinants of oral health and dental visiting patterns. BASIC RESEARCH DESIGN AND PARTICIPANTS: FINRISK senior survey was conducted by the National Public Health Institute in the two areas in 1997. Data were collected through a postal questionnaire, a health examination and an interview, which included questions of number of extracted teeth, dental treatment need and utilisation. The sample consisted of 1,500 persons, 250 women and 500 men aged 65 74 years in both areas; the response rate was 86%. RESULTS: Forty-one per cent of the respondents reported having lost all their teeth and 44% had visited a dentist during the past year. Differences between the areas concerning both dental health and attendance were considerable. Respondents living in North Karelia had lost more teeth and visited a dentist more seldom than respondents in the Helsinki region. Risk indicators for the number of extracted teeth differed between the areas, while determinants for the dental attendance during the previous year were almost identical. The major predictors for a dental visit during the previous year were number of teeth and household income. Self perceived need for dental care was minor. Functional capacity, medically compromising diagnoses or medication did not play any apparent role in dental attendance. CONCLUSIONS: The number of teeth together with income are the principal determinants for dental visits among older persons in Finland. PMID- 11421401 TI - Interrelationship between dental age and chronological age in Somali children. AB - OBJECTIVE: To compare dental age (DA) with chronological age (CA) in Somali and white Caucasian children, resident in Sheffield. DESIGN: Cross-sectional study. SETTING: Paediatric dentistry clinic, Charles Clifford Dental Hospital, Sheffield. SUBJECTS: Somali children under 16 years of age and age- and gender matched white Caucasian subjects. OUTCOME MEASURES: Dental age was determined for each subject, using their existing panoramic radiographs. Comparisons of the difference between dental age and chronological age (DA-CA) were made for gender and ethnic group, using independent sample t tests at the 0.05 level of significance. RESULTS: The sample group comprised 162 subjects: 84 Somali and Caucasian boys (mean age 10.55 years) and 78 Somali and Caucasian girls (mean age 11.24 years). The mean difference between DA and CA was found to be 1.01 years for Somali boys, 0.19 years for Caucasian boys, 1.22 years for Somali girls, and 0.52 years for Caucasian girls. The difference between DA and CA was significantly greater in Somali subjects than in Caucasian children. A few Somali subjects showed a marked discrepancy between chronological age and dental age (range -1.75 to 5.42 years), which was most evident in 8- to 12-year-old children. CONCLUSION: Somali children are significantly more dentally advanced than their Caucasian peers. This finding highlights the need for population specific dental development standards for accurate dental age assessment. PMID- 11421402 TI - The relationship between normative orthodontic treatment need and measures of consumer perception. AB - AIM: The first aim was to investigate whether there was an association between normative (clinician measured) orthodontic treatment need and the following consumer values, 1) child self-perceived aesthetic need, 2) child self-esteem, 3) oral aesthetic subjective impact score (OASIS). The secondary aim was to investigate whether there was an association between child self-esteem and 1) child self-perceived aesthetic need 2) OASIS. STUDY TYPE: Prospective, cross sectional. SAMPLE: A random sample of 439, 11-12 year-old children was selected from schools in Greater Manchester. METHOD: Normative and child self-perceived orthodontic treatment need were measured using the Index of Orthodontic Treatment Need (IOTN). The subjects completed questionnaires to measure their self-esteem (Piers Harris) and OASIS score. RESULTS: Children with higher normative IOTN scores had more negative psycho-social impact from their malocclusion (P<0.001). However, there was no association between clinician IOTN grades and child self esteem. Clinician and child-rated IOTN aesthetic component (IOTN AC) grades were statistically significantly different (P<0.05). Higher child self-esteem scores were associated with lower child rated IOTN AC grades (P<0.05) and lower OASIS scores (P<0.001). CONCLUSIONS: Only one of the consumer measures studies (OASIS) reflected normative/clinician IOTN grades. A high child self-esteem appears to be related to their self-perceived malocclusion and its psycho-social impact. It is still important to have additional information derived from consumer based measures rather than rely solely on normative measures of need. It remains to be seen whether these factors subsequently influence demand and uptake of orthodontic treatment. PMID- 11421403 TI - Epidemiology of malocclusion and orthodontic treatment need of 12-13-year-old Malaysian schoolchildren. AB - OBJECTIVE: Data on malocclusion and orthodontic treatment need in Malaysia are limited. The purpose of this study was to evaluate malocclusion and orthodontic treatment need in a sample of 12-13-year-old schoolchildren using the Dental Aesthetic Index (DAI), and to assess the relationship between malocclusion and socio-demographic variables, perceptions of need for orthodontic treatment, aesthetic perception and social functioning. METHOD: The sampling procedure involved a multistage, clustered and stratified random sampling. The sample comprised of 1,519 schoolchildren attending 20 secondary government and government-aided schools in urban and rural areas of Klang District in Peninsular Malaysia. There were 772 males and 747 females. Each subject was administered a questionnaire eliciting standard demographic information such as gender, parents' income and ethnic origin and questions on perception of need for orthodontic treatment and satisfaction with dental appearance and function. Intra-oral examination for occlusal status using the DAI was performed for each subject. RESULTS: Most subjects (62.6%) require no orthodontic treatment. Only about 7% had handicapping malocclusion that needed mandatory treatment. Malocclusion, as defined in this study, was found to be significantly associated with gender and subjects' area of residence. There were no significant differences in mean DAI scores for Malays, Chinese and Indian children. Significant associations were found between DAI scores and perception of need for orthodontic treatment, satisfaction with dental appearance and social functioning (P<0.01). CONCLUSION: The findings will be useful for the public dental service to determine priority for orthodontic treatment as part of the comprehensive care provided by the School Dental Service (SDS). PMID- 11421404 TI - Matching dental practice board data to the health authority population register. AB - OBJECTIVE: To evaluate the process of matching Dental Practice Board (DPB) registration data to the health authority (HA) register for a defined locality. This included testing for the presence of socio-economic bias in the postcodes held in the DPB data sets and to determine if patients travelling to obtain primary dental care would have an effect on the matching process at health authority level. BASIC RESEARCH DESIGN: All 21 general dental practitioners in Ellesmere Port, South Cheshire gave written consent for the DPB to provide files of their registered patients to the health authority. The registered patients were electronically matched against residents in the HA register in a four stage process. Testing for the presence of a socio-economic bias in the postcodes included in the DPB data set was undertaken by constructing frequency distributions of the proportions of postcoded, and non or partially postcoded DPB records by Super Profiles Lifestyle group. A chi square test was used to test for significant differences. The extent of patients travelling to access primary dental care was assessed by comparing the proportions of registered patients of under 6 years, 25-60 years and those aged 65+ who lived outside South Cheshire Health Authority yet obtained their GDS care within Ellesmere Port. MAIN OUTCOME MEASURES: Out of a total of 39,474 records, 458 (1.2%) were duplicates. Some 35,447 (90.9%) of DPB records could be matched to those contained in the HA register. No significant (chi2 =10.85, df=9, P=0.27) socio-economic difference was found in the proportions of DPB records originally with, and without postcodes by Super Profiles Lifestyle group. Some 1,688 (47.3%) of the unmatched subjects were resident in health authorities other than South Cheshire and therefore could not be expected to be found in the HA register. Nearly 90% of children under 6 years registered with an Ellesmere Port GDP were South Cheshire residents, compared to approximately 85% of adults of working age and older adults. CONCLUSIONS: In this locality DPB data could be matched to the HA register with a reasonable degree of success. However, because of the time taken to clean and make the data sets compatible, the methodology used is inappropriate for routine use. In this locality no socio-economic bias could be found in the records with missing postcodes in the DPB data set. Patients travelling to access dental care would have little influence on the matching process at health authority level in this locality, irrespective of the age group under study. PMID- 11421405 TI - The mouth-body split: conceptual models of oral health and their relationship to general health among ethnic minorities in South Thames Health Region. AB - OBJECTIVE: To identify and describe conceptual models of oral health shared by people from different minority ethnic groups, in particular the relationship between 'oral health' and 'general health'. To identify how these conceptions vary across social factors. BASIC RESEARCH DESIGN: In-depth interviews. Analysis of the interview data to identify the conceptual models used by participants in discussing oral health and its relationship to general health. PARTICIPANTS: Ninety-five individuals from different ethnic groups. RESULTS: Two models of 'health' were identified. The first reflected a traditional model which related health to the absence of disease. The second encompassed a broad definition of health including not only physical but social and psychological well-being and the ability to carry out everyday functions. The model within which an individual operated was influenced by ethnicity and gender. Additionally, two models of the relationship between oral health and general health were identified: one in which the two concepts were seen as separate but related; the other in which oral health and general health were viewed as inseparable aspects of a single dimension. CONCLUSIONS: People from minority ethnic groups vary in their understanding of oral health and its relationship to general health. This variation can be explained in part by ethnic and cultural factors. This finding has important implications both for the design of measures which seek to assess oral health related quality of life, and for the development of health promotional materials. PMID- 11421406 TI - The use of general anaesthesia for orthodontic extractions. AB - AIM: The aims of the study were to evaluate 1) the uptake of general anaesthesia 2) the dental factors influencing use of general anaesthesia for routine orthodontic extractions. RESEARCH DESIGN: Retrospective cross-sectional. PARTICIPANTS: All patients, aged 16 years and under (n=145), currently undergoing orthodontic treatment at the University Dental Hospital of Manchester, who had received extractions, for their current course of treatment. METHOD: Patients were asked to complete a questionnaire after appliance adjustment. MAIN OUTCOME MEASURES: 1) type of anaesthesia received for the most recent orthodontic extractions 2) whether a choice of anaesthesia was given 3) number of permanent teeth removed for the current course of orthodontic treatment 4) previous experience of general anaesthesia. RESULTS: The response was 87%. Twenty-two per cent of patients received general anaesthesia, 64% local anaesthesia and 14% inhalation sedation. Sixty-five per cent of patients were not given a choice of anaesthesia for extractions. Stepwise logistic regression analysis revealed that a patient was more likely to receive a general anaesthetic if: 1) they had received a GA in the past (for fillings or extractions) 2) more than four teeth were extracted. CONCLUSIONS: Local anaesthesia was predominantly used for orthodontic extractions although the use of general anaesthesia was still quite high. The majority of patients were not given a choice of anaesthesia for routine extractions. Previous exposure to general anaesthesia and removal of more than four teeth increased the likelihood of a patient receiving general anaesthesia. PMID- 11421407 TI - The dental caries experience of 5-year-old children in Great Britain. Surveys coordinated by the British Association for the Study of Community Dentistry in 1999/2000. AB - OBJECTIVE: This paper reports the results of standardised clinical caries examinations of 199,440 5-year-old children from across Great Britain. These 1999/2000 coordinated surveys are the latest in a series which seek to monitor the dental health of children and to assess the delivery of dental services. METHOD: The criteria and conventions of the British Association for the Study of Community Dentistry were used. Representative samples were drawn from participating health authorities and boards and caries was diagnosed at the caries into dentine threshold using a visual method without radiography or fibre optic transillumination. RESULTS: The results again demonstrated a wide variation in prevalence across Britain, with mean values for d3mft for the current English regions (of the National Health Service) and for Wales and Scotland ranging from 0.94 in the West Midlands to 2.55 in Scotland. Mean d3mft across Great Britain was 1.57 (d3t=1.14. mt=0.22, ft=0.21). Overall, 40% of children had evidence of dentinal caries experience (d3mft>0), although the means ranged between 30% (West Midlands) and 55% (Scotland). The distribution of caries was highly skewed. Thus the British mean caries experience for those with the disease was 3.88, as opposed to the overall mean of 1.57. Trends over time demonstrate a modest improvement of 4% in overall d3mft for Britain since 1997/98, compared with the 8.6% improvement seen for the two previous years. All three components with dmft have also fallen. The care index has remained virtually unchanged in Britain as a whole (13.6% in 1999/2000, compared to 13.9% in 1997/8). Regional/country means for 1999/2000 ranged from 8-20%. This indicator has not, however, regained the levels seen in the past. CONCLUSION: There has been some improvement in the dental health of 5-year-old children. Overall, the provision of operative care for those with dental decay has not changed; significant groups remain within the population of 5-year-old children who have dental disease and who are in need of dental care. PMID- 11421408 TI - I wish to express my concern about the methods described in the paper by Lo et al. (2000) and the standards of data analysis in that paper. PMID- 11421409 TI - Does dental care improve the oral health of older adults? AB - OBJECTIVE: To assess the relationship between self-perceived change in oral health status and the provision of dental treatment in an older adult population. DESIGN: A longitudinal study with data collection at baseline and after three years. Information on change in oral health was obtained by interviews with study subjects and information on dental treatment over three years was obtained from subjects' dentists. SUBJECTS: Nine hundred and seven subjects took part at baseline and 611 at follow-up. Of the latter, 495 reported at least one dental visit during the three-year observation period and dental treatment information was available for 408. Outcome measures Global transition judgements and change scores derived from four oral health indexes were used to assess change in oral health status. RESULTS: Over the three-year period, one-tenth of subjects reported that their oral health had improved and one-fifth that it had deteriorated. Those who improved made significantly more dental visits and received significantly more dental services that those who deteriorated or did not change (P<0.0001). They also received a broader range of diagnostic, preventive and therapeutic services. The association between change and dental service provision remained after controlling for other potential determinants of oral health. CONCLUSION: The study suggests that improvements in the oral health of older adults depend upon access to comprehensive dental treatments which can address fully their clinical and self-perceived needs. PMID- 11421410 TI - Prospective cohort studies using hydrolysed formulas for allergy prevention in atopy-prone newborns: a systematic review. AB - The aim of this study is to give a systematic overview on publications having investigated the allergy preventive effect of extensively and/or partially hydrolysed infant formulas. Publications were searched by several strategies. Inclusion criteria were: prospective cohort study, randomisation, family history of atopy of the subjects, follow-up for at least the 1st year of life. Studies were described systematically as to methods, study participants, interventions and co-interventions, and study outcomes. The methodological quality was judged according to an instrument assessing the reduction of bias (score between 0 and 7 points). Data of studies having examined partial hydrolysates are presented as odds ratios. Of 16 studies found, 10 fulfilled the inclusion criteria. Study designs showed considerable differences as to methods, intervention and co intervention procedures and definition of outcomes. Methodological quality varied between 0 and 6 points. The comparison of exclusively test- and control formula fed groups showed a uniform tendency towards allergy protection in studies with partial hydrolysates. Study results on the effect of extensive hydrolysates were not comparable because of major differences in study designs. CONCLUSION: More well-designed studies with an adequate statistical power are needed to compare the allergy preventive effect of partially and extensively hydrolysed formulas with a standard infant formula. PMID- 11421411 TI - High-birth weight and other risk factors for Wilms tumour: results of a population-based case-control study. AB - Wilms tumour, or nephroblastoma, is one of the childhood cancers included in two recent population-based case-control studies in West Germany. Altogether, 177 children under the age of 10 years with Wilms tumour diagnosed between 1988 and 1994 and 2006 control children sampled from population registration files participated. Information on potential risk factors was obtained from the parents using a questionnaire and by subsequent telephone interview. We found an association with a high birth weight >4000 g (odds ratio 1.58; 95% confidence interval 1.01-2.48), which was somewhat stronger for children aged 2 years or older. Findings for young maternal age at birth and certain parental occupationally related exposures were not reported by previous studies and thus may be chance findings. As opposed to previous studies, we failed to confirm associations with high parental age at birth, maternal coffee and tea consumption during pregnancy, and exposure to pesticides. CONCLUSION: Based on this large population-based case-control study, high birth weight may play a role in the aetiology of Wilms tumour, but many risk factors previously suggested are of less importance. PMID- 11421412 TI - Deciding on treatment limitation for neonates: the parents' perspective. AB - The reported study aimed to explore parents' perceptions of treatment withdrawal/withholding; their experience and their opinions. In-depth face-to face interviews at 3 and 13 months after the baby's death were conducted with 108 parents of 62 babies from the East of Scotland. Of the parents, 56% (60/180) believed they had decided to stop treatment. A further 4/59 sets of parents subsequently wished they had taken responsibility. Only one parent who did decide felt guilt 3 months after the event. Parents identified two essential factors which minimise doubt: full and honest information and concrete evidence of a poor prognosis. CONCLUSION: The majority of parents wish to be active in decision making on behalf of their baby. Doing so does not appear to have adverse consequences. The pacing of events in the process of deciding and managing the dying is critical. Dissatisfaction is reduced if parents are given time and evidence to help them assimilate the reality at each stage. PMID- 11421413 TI - Plasma levels of interleukin-6 and interleukin-10 in preterm neonates evaluated for sepsis. AB - In a prospective study, plasma interleukin-6 (IL-6) and interleukin-10 (IL-10) levels were measured by enzyme-linked immunosorbent assay in 45 premature neonates (25-34 weeks gestational age) with signs and symptoms of suspected sepsis at 0, 12 and 24 h; C-reactive protein (CRP) was measured at 0-24 h after enrolment. Six subjects were excluded due to insufficient blood sampling. The remaining 39 neonates were assigned to one of three groups: 25 newborns with sepsis (blood culture positive), seven with pneumonia (positive results on broncho-alveolar lavage fluid culture and characteristic chest radiography) and seven with necrotising enterocolitis (NEC) (characteristic intestinal and radiological signs according to the criteria of Bell et al.). A group of 20 healthy preterm neonates represented control subjects. On admission, higher levels of IL-6, IL-10 and CRP were observed in neonates with sepsis: IL-6 (median 1500 pg/ml, range 487-10000 pg/ml), IL-10 (median 113 pg/ml, range 70-196 pg/ml), CRP (median 22 mg/l, range 4-80 mg/l); pneumonia: IL-6 (median 1500 pg/ml, range 747-8000 pg/ml, IL-10 (median 84 pg/ml, range 76-92 pg/ml), CRP (median 10 mg/l, range 8-33 mg/l) and NEC: IL-6 (median 6650 pg/ml, range 1595-7950 pg/ml), IL-10 (median 80 pg/ml, range 61-147 pg/ml), CRP (median 3 mg/l, range 2.8-8 mg/l) as compared to controls (IL-6 median 208 pg/ml, range 198-349 pg/ml; IL-10 median 36 pg/ml, range 19-50 pg/ml; CRP median < 2 mg/l) (P < 0.05). In neonates with sepsis, IL-6 levels were significantly correlated with IL-10 levels (r = 0.65; P = 0.04) at the time of the second sample. The highest IL-6 levels were observed at onset, while IL-10 was predominant 12 h later. On admission, IL-10 and CRP levels were significantly higher in non-survivors (IL-10 median 507 pg/ml, range 422-753 pg/ml; CRP median 123 mg/l, range 20-219 mg/l) than in survivors (IL-10 median 76 pg/ml, range 61-143 pg/ml; CRP median 8 mg/l range 3-46 mg/l), while IL 10 levels were significantly higher (P < 0.05) also 12 h after admission (non survivors: IL-10 median 600 pg/ml, range 538-800 pg/ml; survivors: IL-10 median 74 pg/ml, range 53-161 pg/ml). IL-6 and IL-10 levels were significantly correlated with CRP levels on admission (r = 0.45; P = 0.05). CONCLUSION: Preterm neonates with sepsis, pneumonia or necrotising enterocolitis showed increased interleukin-6, interleukin-10 and C-reactive protein levels. High interleukin-10 concentration was associated with mortality and could be an early indicator of prognosis. PMID- 11421414 TI - Human herpes virus type 7 DNA in the cerebrospinal fluid of children with central nervous system diseases. AB - Human herpes virus type 7 (HHV-7) has been associated with unspecific febrile syndrome, exanthem subitum (ES), viral rashes and Epstein-Barr virus (EBV) like syndrome. Neurological complications such as hemiplegia or seizures have been described in a few children with ES. Whether HHV-7 may also affect the CNS in the absence of ES is unknown. In this study, we investigated CSF samples from children with different neurological diseases for the presence of HHV-7 specific DNA. A HHV-7 specific nested polymerase chain reaction (PCR) was established amplifying a 478 bp DNA sequence of the glycoprotein U23 of HHV-7 strain SB. 68 children with CNS diseases with inflammatory CSF findings (n = 24), CNS diseases without inflammatory CSF findings (n = 18) and febrile seizures (n = 26) were examined. A total of 26 children with infectious diseases in the absence of neurological disease and 11 children without signs of a peripheral infection and without neurological disease served as controls. The CSF samples of six children from the study groups were HHV-7 PCR positive, but none from the controls. These children were diagnosed with aseptic meningitis (n = 1), viral encephalitis/meningoencephalitis (n = 2), facial palsy (n = 1), vestibular neuritis (n = 1) and febrile seizure (n = 1). CONCLUSION: These results indicate that human herpes virus type 7 infection is associated with central nervous system disease in children and should be considered in children whether inflammation in the cerebrospinal fluid is present or not. PMID- 11421415 TI - Growth hormone treatment enhances bone mineralisation in children with chronic renal failure. AB - Dual energy X-ray absorptiometry of the whole body and the lumbar spine was performed to study bone mineralisation before and after 1 year of recombinant human growth hormone (rhGH) treatment in ten children with chronic renal failure. At the start, median age was 7.3 years (range 2.0-8.8 years) and median glomerular filtration rate 15 ml/min per 1.73 m2 (range 7-41 ml/min per 1.73 m2). Total body mineral content (TBMC), lumbar spine mineral content (LBMC), total body bone mineral density (TBMD) and lumbar spine mineral density (LBMD) improved significantly (P < 0.05) after 1 year of treatment. Bone mineral data before and after treatment were compared with two groups of controls, i.e. ten healthy children matched for age and ten healthy children matched for height. Patients' TBMC, LBMC, TBMD and LBMD data before treatment were no different from those of height-matched controls; the same was true after 1 year of treatment except for the patients' significantly better LBMD (P < 0.05). When compared with age matched controls, patients had significantly lower baseline TBMC and LBMC levels before treatment; after treatment LBMC was no longer different. However, there were no differences in TBMD or LBMD between patients and age-matched controls at baseline or after rhGH. CONCLUSION: Recombinant human growth hormone treatment for 1 year results in a significant increase in both growth velocity and bone mineralisation. Comparison with height-matched controls shows a similar bone mineralisation at baseline and a better bone mineral density after treatment. PMID- 11421416 TI - Ethical dilemmas in neonatology: recommendations of the Ethics Working Group of the CESP (Confederation of European Specialists in Paediatrics). AB - Neonatal intensive care has greatly improved the survival chances of a very sick infant. At the same time, it has also given rise to serious ethical problems. In all circumstances, however, parents and paediatricians and other healthcare team workers should continuously evaluate together what is in the best interest of the infant and react accordingly. It is also clear that the principle "the best interests of the infant" can be interpreted in different ways; therefore no simple guideline is possible. PMID- 11421417 TI - Colonic pneumatosis intestinalis in preterm infants: different to necrotising enterocolitis with a more benign course? AB - Necrotising enterocolitis (NEC) is the predominant immaturity-associated disease of the bowel in the preterm neonate and leads to substantial mortality and long term morbidity. Diagnostic features of NEC include, apart from the clinical presentation, laboratory and radiological parameters. Pneumatosis intestinalis (PI) on abdominal radiographs is regarded as a criterion of definitive proof for this disease entity. The objective of this presentation is to link the topographic pattern of PI to the clinical course in cases of suspected NEC. We report two cases of radiological appearance of colonic PI indicative of NEC without associated laboratory and/or macroscopic evidence of the disease, and with minor clinical symptoms only. Data from animal studies indicate that decreased blood supply to the small bowel is associated with more extensive bowel damage as compared to impaired colonic perfusion. Therefore the topographical distribution of PI might be a more specific sign as the general presence or absence of PI. CONCLUSION: The topographical pattern of pneumatosis intestinalis may be predictive of the clinical course of inflammatory bowel disease in preterm infants. PMID- 11421418 TI - Cerebellar defect associated with Schimke immuno-osseous dysplasia. AB - We report the finding of an absent cerebellar hemisphere and partial absence of the cerebellar vermis in a child with dysmorphic features, spondyloepiphyseal dysplasia, steroid resistant nephrotic syndrome secondary to focal segmental glomerulosclerosis and T-cell lymphopenia (Schimke immuno-osseous dysplasia). These findings have not, to our knowledge, been described before and are likely to represent the consequence of a vascular event either in-utero or in early infancy. CONCLUSION: Cerebral imaging should be performed early in the course of the disease and should be repeated if further neurological events develop. PMID- 11421419 TI - Familial polythelia over three generations with polymastia in the youngest girl. AB - Supernumerary nipples or polythelia are developmental abnormalities located along the embryonic mammary lines. It is the most common form of accessory breast tissue malformation and usually occurs sporadically but familial aggregation has been reported. Polythelia has been reported in association with congenital malformations, in particular with renal anomalies. Polymastia in female patients has been reported to manifest usually during pregnancy or lactation. We report on a pedigree with six cases of polythelia over three generations and one case of polythelia and polymastia in the youngest member of this family. The girl (11 years old ) had in addition to six supernumerary nipples, an accessory breast gland located under the normal left breast. No other congenital malformations could be detected. This girl will remain under follow-up until the end of puberty when the accessory breast gland will be removed. Manifestation of polymastia during puberty rarely has been reported. CONCLUSION: Polymastia may appear with familial polythelia even without renal anomalies. PMID- 11421420 TI - Severe and prolonged sedation in five neonates due to persistence of active diazepam metabolites. AB - Five neonates who suffered from an unexpected long period of respiratory failure, muscular hypotonia, and drowsiness were observed in a retrospective study. Prior to this general depression, unusually high doses of diazepam were administered to all patients via intravenous bolus injection. Serum levels of diazepam and its active metabolites were substantially elevated in the course of the disease. The persistence of the very long-acting N-desmethyldiazepam with considerable extension in neonates and even more exaggerated in premature infants is emphasized due to the reduced capacity of the hepatic biotransformation system. CONCLUSION: The intravenous application of diazepam imposes a risk of marked and prolonged general depression in neonates. Pronounced adverse effects are to be expected for prematures even after a single diazepam intravenous bolus if the dosage is not appropriate. Diazepam should not be used for short sedation and is not the drug of choice for anticonvulsant therapy in neonates. PMID- 11421421 TI - Protein malnutrition due to replacement of milk by rice drink. AB - We present the case of an 8-month-old boy with failure to thrive and hypoalbuminaemia due to protein malnutrition caused by the replacement of milk by a rice drink. All symptoms resolved with an appropriate feeding. CONCLUSION: Rice drinks are no alternatives to infant milk formulas and carry a serious risk for the development of protein malnutrition. PMID- 11421422 TI - Nosocomial necrotising enterocolitis outbreaks: epidemiology and control measures. AB - Necrotising enterocolitis (NEC) is one of the most serious gastrointestinal diseases among newborns and it mainly affects those in intensive care units. The aetiology of the disease has been reported to be multifactorial and both sporadic cases and nosocomial outbreaks have occurred. In this report, we review 17 epidemics of NEC reported in the literature between 1973 and 1999. The number of confirmed cases ranged from 1 to 32 with an average of 10.5 confirmed cases. On average, 16.15% of cases required surgery (range 0-66.6%). The average mortality rate was 6.25% (range 0-87.5%). The mean age at disease onset was 9.5 days (range 6.6-29 days). Most of the infants had low birth weight (median weight 1,395 g; range 1,112-2,788 g, calculated on the reported mean weights). The main risk factors associated with NEC were: low birth weight, low gestational age, low Apgar score, perinatal complications, hyaline membrane disease, and umbilical catheterisation. The bacteria involved often included Enterobacteriaceae, particularly Escherichia coli, Klebsiella pneumoniae and Enterobacter cloacae type 3305573. The causative role of Clostridia in NEC is controversial. With regard to viral agents, coronarovirus, rotavirus and enterovirus, such as echovirus type 22, were isolated during some of the epidemics. The recommended control measures for NEC epidemics are those used for epidemics of other orofaecally transmitted infections. CONCLUSION: Understanding the epidemiology of necrotising enterocolitis is fundamental if adequate preventive control measures are to be developed and applied. PMID- 11421423 TI - Jugular venous malformation in an 8-year-old boy: treatment with endovascular sclerotherapy. AB - An 8-year-old male presented with a mass in the left supraclavicular region first noted 3 months earlier and which gradually became more prominent. Ultrasound showed a lobular, well-delineated hypoechoic lesion which increased in size on Valsalva manoeuvre. Doppler waveform analysis suggested a slow flow vascular lesion. Venography showed a saccular, multilobular venous malformation which connected with the external jugular and subclavian veins. With an angiographic catheter, the venous malformation was treated by endovascular sclerotherapy. Four weeks later, ultrasonography showed a resolution of the lesion. CONCLUSION: Endovascular sclerotherapy appears to be an effective and safe treatment for jugular venous malformation. PMID- 11421424 TI - A small-for-dates newborn girl with a mummified left arm. PMID- 11421425 TI - Is low serum complement 3 in Henoch Schonlein purpura unusual and of prognostic value? PMID- 11421426 TI - Varicella zoster virus infection complicated by Neisseria meningitidis bacteraemia in two children. PMID- 11421427 TI - Isolated 6th nerve palsy in a child associated with asymptomatic Chlamydia pneumoniae infection and elevated anti-GQ1b antibody. PMID- 11421428 TI - Neuroleptic malignant syndrome following withdrawal of amantadine in a patient with influenza A encephalopathy. PMID- 11421429 TI - Primary congenital lymphoedema with unilateral arm involvement. PMID- 11421430 TI - Prehospital thrombolysis. PMID- 11421431 TI - Community thrombolysis in the Coromandel region. Audit of the "Cardiac Events in the Coromandel-Assessment Strategy and Triage" (CE-COAST) pilot program. AB - AIM: To audit the experience of a pilot program for community thrombolysis undertaken within the Coromandel region. METHODS: Community thrombolysis for patients suffering acute myocardial infarction (MI) was undertaken in areas within the Coromandel peninsula greater than half an hour by road from Thames Hospital. Thrombolytic therapy (Retelapse) was given following a discussion and review of a digitally transmitted ECG with the cardiology registrar. Treatment times and patients demographics were prospectively recorded. Subsequent clinical events were by chart review. Comparison of treatment times were made with an historical cohort for the same population which had received in-hospital thrombolysis between 1993 and 1998. RESULTS: Between July 1998 and December 1999, nineteen patients received thrombolysis in the community. There were no arrhythmic events during transportation and no deaths or reinfarctions during hospital stay. Median time from pain onset to thrombolysis was 135 (mean 175.5 +/ 144.9 SD) minutes which equated to a reduction in median time delay of 135 minutes compared to that experienced by the historical cohort (median 270, mean 316.7 +/- 145.8 SD minutes), p=0.0003. CONCLUSION: Community thrombolysis is logistically feasible within the New Zealand setting and results in major time reductions in the treatment of patients with acute MI. PMID- 11421432 TI - Adverse events regional feasibility study: methodological results. AB - AIMS: To assess the feasibility of research into the occurrence, causation and prevention of adverse events (AEs) in New Zealand public hospitals. METHODS: A two-stage retrospective review was carried out on 1,575 medical records selected by systematic list sample from admissions for 1995 in three public hospitals in the Auckland region. Following initial screening, medical records were subject to structured implicit review using a standardised protocol. Feasibility measures, using international benchmarks where possible, were: adequacy of sample selection; completeness of medical records; reliability and validity of screener and reviewer judgements; internal consistency and face validity of AE determination and preventability assessment. RESULTS: The sample selection procedure was effective, although nearly 10% of records could not be secured. Information in medical records was sufficient for the identification and analysis of AEs. Adequate levels of agreement were achieved for screener and reviewer judgements, with kappa scores ranging between 0.302 and 0.622 and positive predictive values between 50.0% and 89.7%. The criteria for AE determination showed internal consistency and face validity, as did those for preventability. CONCLUSIONS: Research into the occurrence, causation and prevention of AEs in New Zealand health care settings is methodologically feasible and meets international benchmark standards. PMID- 11421433 TI - Adverse events regional feasibility study: indicative findings. AB - AIMS: To identify substantive findings of potential clinical and managerial significance from a regional feasibility study of adverse events (AEs). METHODS: A standardised protocol using structured implicit review was applied to 142 AEs generated in an audit study of three public hospitals in the Auckland region for admissions in 1995. Areas of potential significance addressed were: timing, location and impact of AEs; preventability; and clinical context and predictability. RESULTS: 142 cases were identified as AEs (10.7% of 1,326 screened records). In 102 cases, 7.7% of all screened records, it was considered to be more likely than not that health care management contributed to the AE. About half the reported AEs occurred before the index admission, the majority outside hospital. Over half of all events resulted in disability that was resolved within a month. An average 6.7 extra days stay in hospital were attributable to AEs. For 60% of AEs the evidence for preventability was either low or nonexistent. Areas of potential prevention were predominantly educational. Over half of all AEs occurred in a surgical context. Medical AEs were more likely to have occurred outside hospital, to be drug-related, to be associated with an acute admission, to be classified as highly preventable, and to have a greater impact on hospital stay. CONCLUSIONS: Although the data generated by a feasibility study must be treated with caution, the pattern of results is consistent with comparable Australian findings and is of potential clinical and managerial significance. PMID- 11421434 TI - The community services card: does it make a difference to pharmaceutical utilisation? AB - AIMS: To estimate the utilisation rate, and amount of state subsidy of prescription items per head by age, sex and community Services Card (CSC) for the year ended June 1999. METHODS: Data from a market research company (IMS Health), Health Benefits Limited, Statistics New Zealand and Work and Income New Zealand were used to calculate average per head per year pharmaceutical utilisation rate and subsidy cost for CSC holders and non-holders. RESULTS: For both sexes, and for all age groups, CSC-holders tended to use more prescription items per head and incur higher subsidy cost than non-holders. The standardised CSC utilisation rate was 2.6 times the non-CSC rate. For children, average per-item subsidy cost for CSC-holders was lower than for non-holders; the reverse was true for adults. CONCLUSION: CSC holders had higher pharmaceutical utilisation rates than non holders at a national level (but not necessarily at a local level). If non-uptake of cards and health status were taken into account, however, it is possible that pharmaceutical utilisation rates were suboptimal amongst those most in need of services. Analyses are urgently required to examine prescribing patterns at a regional level. PMID- 11421435 TI - Heart disease and diabetes risk factors in Pacific Islands communities and associations with measures of body fat. AB - AIMS: To describe the prevalence of obesity and other coronary heart disease and Type 2 diabetes risk factors by age and ethnic group in Pacific Island communities and to determine the associations between these risk factors and body mass index. METHODS: Cross-sectional data from commuity-based intervention projects were combined to provide anthropometric, blood sample and blood pressure data on 1,175 Pacific Islands people (467 men, 708 women) aged 20 years and over from church communities in South, Central and West Auckland. Self-reported data on diabetes status and leisure-time physical activity were also collected. RESULTS: Based on an ethnic-specific mass index (BMI) cut-off (> 32 kg/m2), 45% of men and 66% of women were obese. The age-standardised prevalence of known diabetes was 12%. Men and women aged 40-60 years had the highest risk factor levels and were the most sedentary. Tongans had higher risk factor levels than Samoans. In men, BMI and waist circumference were associated (p<0.05), in the direction of greater disease risk, with blood pressure and concentrations of total cholesterol, HDL-cholesterol, LDL-cholesterol, triglycerides, and blood glucose. In women, these associations were similar but less consistent. CONCLUSIONS: While these data are not representative for all Pacific people living in New Zealand, they do show an extremely high prevalence of obesity and significant associations between obesity and other cardiovascular risk factors. These communities warrant a very high priority as part of public health efforts to address New Zealand's growing obesity epidemic. PMID- 11421436 TI - Should decisions on treatment be based on absolute benefit rather than absolute risk? PMID- 11421437 TI - Recommending particular treatment options: the vitamin K experience. PMID- 11421438 TI - Unethical to not offer voluntary HIV screening to all pregnant women. PMID- 11421439 TI - Problems with assessing the value of D-dimer for excluding deep vein thrombosis. PMID- 11421440 TI - Ethnicity and poverty are not deterrents to high childhood immunisation rates. PMID- 11421441 TI - Facilitating global medical registration. PMID- 11421442 TI - Patient safety: a fence at the top of the cliff. PMID- 11421443 TI - Clinical risk management--patient safety and informed consent. PMID- 11421444 TI - Paraquat resistance of weeds--the case of Conyza canadensis (L.) Cronq. AB - The paper gives an overview of literature on paraquat resistance of weeds and the proposed mechanism of resistance. New results we achieved on horseweed (Conyza canadensis /L./, Cronq.) are discussed in detail. It was demonstrated that there is no significant constitutive difference related to the paraquat resistance between untreated susceptible and paraquat-resistant horseweed plants. The lower sensitivity of flowering resistant plants may be due to the fact that paraquat content in treated leaves of flowering resistant plants was only 25% as compared to those measured at rosette stage. Our results confirm that paraquat resistance is not based on elevated level and activity of antioxidant enzyme system. The hypothesized role of polyamines in the resistance mechanisms can be excluded. The higher putrescine and total polyamine content of paraquat treated resistant leaves can rather be regarded as a general stress response, than as a symptom of paraquat resistance. A paraquat-inducible protein is supposed to play a role in the resistance, which presumably functions by binding paraquat to an inactivating site and/or by carrying paraquat to metabolically inactive cell compartment (vacuole, cell wall). From model experiments it is concluded that paraquat and diquat preferentially form hydrophylic interactions with proteins containing a higher amount of lysine and glutamic acid. Consequently, the reason for paraquat resistance in horseweed is probably a hydrophylic interaction of paraquat with a protein, leading to inactivation of paraquat through forming a conjugate and/or sequestration into the vacuole or the cell wall. PMID- 11421445 TI - Lignan and coumarin glycosides from Haplophyllum suaveolens. AB - Two new lignan glycosides of dibenzylbutyrolactol type (haplomarin) and arylnaphthalene type (haploborin) were isolated from the aerial parts of Haplophyllumn suaveolens together with the known arylnaphthalene lignan glycoside arabelline and hydroxycoumarin glycoside xeroboside. So far arabelline is found only in H. buxbaunii. This is the first report of the isolation of xeroboside from Haplophyllum. PMID- 11421446 TI - Tricarboxylic acid cycle enzymes of the ectomycorrhizal basidiomycete, Suillus bovinus. AB - In crude cell extracts of the ectomycorrhizal fungus, Suillus bovinus, activities of citrate synthase, aconitase, isocitrate dehydrogenase, succinate dehydrogenase, fumarase, and malate dehydrogenase have been proved and analyzed. Citrate synthase exhibited high affinities for both its substrates: oxaloacetate (Km = 0.018 mM) and acetyl-CoA (Km = 0.014 mM). Aconitase showed better affinity for isocitrate (Km = 0.62 mM) than for citrate (Km = 3.20 mM). Analysis of isocitrate dehydrogenase revealed only small maximum activity (60 nmol x mg protein(-1) x min(-1)), the enzyme being exclusively NADP+-dependent. Using the artificial electron acceptor dichlorophenol indophenol, activity and substrate affinity of succinate dehydrogenase were rather poor. Fumarase proved Fe2+ independent. Its affinity for malate was found higher (Km = 1.19 mM) than that for fumarate (Km = 2.09 mM). High total activity of malate dehydrogenase could be separated by native PAGE into a slowly running species of (mainly) cytosolic (about 80%) and a faster running species of (mainly) mitochondrial origin. Affinities for oxaloacetate of the two enzyme species were found identical within limits of significance (Km = 0.24 mM and 0.22 mM). The assumed cytosolic enzyme exhibited affinity for malate (Km = 5.77 mM) more than one order of magnitude lower than that for oxaloacetate. FPLC on superose 12 revealed only one activity band at a molecular mass of 100 +/- 15 kDa. Activities of 2-oxoglutarate dehydrogenase and of succinyl-CoA synthetase could not be found. Technical problems in their detection, but also existence of an incomplete tricarboxylic acid cycle are considered. Metabolite affinities, maximum activities and pH dependences of fumarase and of malate dehydrogenase allow the assumption of a reductive instead of oxidative function of these enzymes in vivo. PMID- 11421447 TI - Betalains and phenolics in red beetroot (Beta vulgaris) peel extracts: extraction and characterisation. AB - The extraction of red beetroot (Beta vulgaris) peel betalains and phenolics was compared with two extraction methods and solvents. The content of total phenolics in the extracts was determined according to a modification of the Folin-Ciocalteu method and expressed as gallic acid equivalents (GAE). The profiles of extracts were analysed by high-performance liquid chromatography (HPLC). The compounds of beetroot peel extracted with 80% aqueous methanol were characterised from separated fractions using HPLC- diode array detection (HPLC-DAD) and HPLC electrospray ionisation-mass spectrometry (HPLC-ESI-MS) techniques. The extraction methods and the choice of solvent affected noticeably the content of individual compounds in the extract. The betalains found in beetroot peel extract were vulgaxanthin I, vulgaxanthin II, indicaxanthin, betanin, prebetanin, isobetanin and neobetanin. Also cyclodopa glucoside, N-formylcyclodopa glucoside, glucoside of dihydroxyindol-carboxylic acid, betalamic acid, L-tryptophan, p coumaric acid, ferulic acid and traces of unidentified flavonoids were detected. PMID- 11421448 TI - Aminophosphonate-induced changes of betacyanine and ionic efflux. AB - Betacyanine and ionic leakage from red beet (Beta vulgaris ssp. L. rapacea) roots and lilac (Syringa vulgaris L.) leaves under the influence of new aminophosphonates were studied by spectroscopic and conductometric methods. It was found that the leakage of dye or electrolytes depended both on the concentration of the compounds used and their structural features. The results compared to those obtained for the well known herbicide Buminafos (dibutyl 1 butylamino-1-cyclohexanephosphonate) enabled to conclude that some of the compounds studied exhibited comparable or better activity than this herbicide. That makes them potentially good herbicides. It is possible that the effects observed are the result of action on cell membranes of the tissues used. The possible role of the structural features of aminophosphonates in this action is discussed. PMID- 11421449 TI - Phytochemical analysis of Phillyrea latifolia L., a new source of oleuropeoside. AB - As a part of our studies on the biologically active substances from Spanish plants, we have undertaken an investigation of the chemical constituents of a typical mediterranean species, Phillyrea latifolia L. (Oleaceae). Two secoiridoid glycosides, three phenylpropanoid glycosides, one lignane and two triterpenic acids were isolated from the leaves of this species and identified. The phytochemical analysis of the aerial parts of P. latifolia revealed that it is a rich source of oleuropeoside. PMID- 11421450 TI - Seasonal variation of the chemical constituents from Croton species. AB - The terpenes, sterols, alkaloid (glaucine) and alpha-tocopherol show seasonal variation for Croton hemiargyreus hemiargyreus and Croton echinocarpus. The amounts of triterpenes are higher during the tropical summer and in most samples the major sesquiterpene was characterized as caryophyllene. The seasonal variation of glaucine showed a maximum between June and October for C. hemiargyreus, and was present only in January and June in C. echinocarpus. PMID- 11421451 TI - Wavelength shifts correlation between near infrared and ultraviolet regions of the LHII bacteriochlorophyll spectrum from Ectothiorhodospira sp. AB - Bacteriochlorophyll a has a maximum at 258 nm previously related to the ring E ester system interacting with the pi-system of the macrocycle. In this work, we compared the effect of lauryldimethylamine-N-oxide (LDAO) and alkaline pH on both the near infrared and the ultraviolet region of the LHII spectrum from Ectothiorhodospira sp. While LDAO induces only a shift of the 850 nm band arising from the Qy transition of the bacteriochlorophyll a, alkaline pH also causes a concomitant and reversible 10-nm shift from 258 to 248 nm. Both shifts have similar apparent pKs (12.3 and 12.6, respectively). Interestingly, the presence of NaCl reduces these pKs values to 11.4 and 11.7. PMID- 11421452 TI - Effect of membrane fluidity on photoinhibition of isolated thylakoids membranes at room and low temperature. AB - The relationship between thylakoid membrane fluidity and the process of photoinhibition at room and low (4 degrees C) temperature was investigated. Two different membrane perturbing agents--cholesterol and benzylalcohol were applied to manipulate the fluidity of isolated pea thylakoids. The photochemical activity of photosystem I (PSI) and photosystem II (PSII), polarographically determined, were measured at high light intensity for different time of illumination at both temperatures. The exposure of cholesterol- and benzylalcohol-treated thylakoid membranes to high light intensities resulted in inhibition of both studied photochemical activities, being more pronounced for PSII compared to PSI. Time dependencies of inhibition of PSI and PSII electron transport rates for untreated and membranes with altered fluidity were determined at 20 degrees C and 4 degrees C. The effect is more pronounced for PSII activity during low-temperature photoinhibition. The data are discussed in terms of the determining role of physico-chemical properties of thylakoid membranes for the response of photosynthetic apparatus to light stress. PMID- 11421453 TI - Transformation of Catalpa ovata by Agrobacterium rhizogenes and phenylethanoid glycosides production in transformed root cultures. AB - Transformed root cultures of Catalpa ovata were established following shoots infection with four agropine strains of Agrobacterium rhizogenes. Frequency of root formation was dependent on the bacterial strain and the presence of acetosyringone in the incubation medium. It is the first report concerning the possibility of transforming Catalpa ovata by A. rhizogenes. Both transformed and untransformed root cultures of C. ovata were studied for their growth and phenylethanoid glycoside production. As with the roots of intact plants, cis- and trans-verbascoside as well as martynoside were produced in transformed and untransformed root cultures of C. ovata. In hairy roots, total (cis + trans) verbascoside production could be stimulated up to three-fold of that of roots of 6-month-old plants grown in a greenhouse, by using an appropriate root line cultured in liquid 1/2 B5 Gamborg medium containing indole-3-butyric acid (0.1 mg/l) in the dark but not light conditions. Transformed and untransformed root cultures of C. ovata were also found to have 10 times higher martynoside production than roots of intact plants. PMID- 11421454 TI - Acetylcholinesterase and insect growth inhibitory activities of Gutierrezia microcephala on fall armyworm Spodoptera frugiperda J.E. Smith. AB - From the aerial parts of Gutierrezia microcephala (Asteraceae), four oxyflavones were isolated, namely 5,7,2'-trihydroxy-3,6,8,4',5'-pentamethoxyflavone (1); 5,7,4'-trihydroxy-3,6,8-trimethoxyflavone (2); 5,7,2',4'-tetrahydroxy-3,6,8,5' tetramethoxyflavone (3); 5,2'-dihydroxy-3,6,7,8,4',5'-hexamethoxyflavone (4), and an ent-clerodane, bacchabolivic acid (5). Compounds 1-5, the synthetic methyl ester (6), n-hexane and MeOH extracts were evaluated against the fall armyworm (Spodoptera frugiperda). Gedunin, a known insect growth regulator isolated from Cedrela spp. was used as a positive control. When tested for activity on neonate larvae into the no-choice artificial diet bioassay, flavone (1), clerodane (5), its methyl ester (6), MeOH and n-hexane extracts caused significant larval mortality with MC50 of 3.9, 10.7, 3.46, 7.95 and 7.5 ppm at 7 days, respectively, as well as growth reduction. They also increased the development time of surviving larvae and a significant delay in time to pupation and adult emergence. Acute toxicity against adults of S. frugiperda was also found, 5, 6, gedunin and n-hexane extract had the most potent activity with LD50 value of 6.59, 15.05, 10.78, and 12.79 ppm, respectively. In addition, MeOH, n-hexane extracts, 5, 6 and gedunin caused acetylcholinesterase inhibition with 93.7, 100, 90.2, 62.0 and 100% at 50.0 ppm, respectively; whereas 1-4 exhibited only moderate inhibitory activity. Compounds 1, 5 and 6 showed inhibitory activities comparable with gedunin. These compounds could be responsible of the insect growth inhibitory activity of this plant. PMID- 11421455 TI - Organometallics and quaternary ammonium salts affect calcium ion desorption from lecithin liposome membranes. AB - The objective of the present work was to compare the effects of groups of tin and lead organometallic compounds and their mixtures with amphiphilic quaternary ammonium salts (QAS) on the process of calcium ion desorption from lecithin liposome membranes, as dependent on the properties of the hydrophilic and hydrophobic parts of QAS. In the investigations the method of radioactive labels was applied. Synergism and antagonism in the action of both groups of compounds were found. The effectiveness of the cooperation depended more on chain length of QAS compounds than on the size and polarity of their hydrophobic parts. The most effective of all compounds studied was a the mixture of benzyldimethylammonium chloride in a mixture with tripropyltin. Since the rate of calcium desorption proved to be a good measure of efficacy of biologically active surfactants, it seems that the conclusions reached in this paper may be useful for choosing compounds which are able to decontaminate the environment polluted with heavy metals. PMID- 11421456 TI - Computer simulations of the electric interactions between the phospholipid head groups and ionic admixtures in the membrane surface. AB - Some phospholipids (e.g. lecithin) form a system of electric dipoles on the membrane surface layer. In the case of lecithin the positive dipole charge is located on the choline and the negative one on the phosphoric molecule group. These dipoles are arranged almost parallel to the membrane surface. Taking the dipole membrane structure as a base for further investigations, a computer model of the electrostatic interaction between the dipole system and the ionic admixture was investigated. The model presumes hexagonal centered or a rectangular flat geometry of the 121 dipoles distribution. The dipoles may rotate freely around round the motionless symmetry axis perpendicular to the system surface. The initial state is given by fixing the geometry of the dipole matrix and ionic admixture distribution. Subsequently this system underwent a computer simulation which consisted of a calculation of resultant force moments acting on each dipole caused by other dipoles and ions. These force moments lead the system to the equilibrium state (minimum of the binding energy). The minimum energy value of the dipoles system depends on concentration and charge of the admixtured ions. The results of repeated simulations indicate that the system achieve the least of all binding energy (the most stable equilibrium state) at 1.5% concentration of admixtured ions in case of ion charge equal to 1Q (where Q denotes arbitrary unit of ion charge) and at 2.5% concentration of admixtured ions in case of ion charge equal to 2Q. The calculated results are in a good agreement with the experimental. PMID- 11421457 TI - Counterion effects on interaction of amphiphilic quaternary ammonium salts with model membranes. AB - The micellization as well as the interaction with model membranes of dodecyltrimethylammonium halides (DTAX) and N-dodecyl-N,N-dimethyl-N benzylammonium halides (DBeAX) were studied at 298K and 313K by means of titration calorimetry. The calorimetric curves reflect both the counterion and benzyl group effects on the interaction of the surfactants studied with the lipid bilayer. Bromide as counterion enhanced the interactions more than chloride of both DTAX and DBeAX compounds with model membranes. Further, we studied the influence of DTAX and DBeAX on calcium ion desorption from the liposome membrane using a radioactive tracer method. DBeAX proved more efficient in desorption of calcium than DTAX. Iodides of these compounds enhanced this process more than bromides and chlorides. PMID- 11421458 TI - Site-directed mutagenesis of proline-285 to leucine in Cephalosporium acremonium isopenicillin-N-synthase affects catalysis and increases soluble expression at higher temperatures. AB - The conversion of delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (ACV) to isopenicillin N is dependant on the catalytic action of isopenicillin N-synthase (IPNS), an important enzyme in the penicillin and cephalosporin biosynthetic pathway. One of the amino acid residues suggested by the Aspergillus nidulans IPNS crystal structure for interaction with the valine isopropyl group of ACV is proline-283. Site-directed mutagenesis of the corresponding proline-285 to leucine in Cephalosporium acremonium IPNS resulted in non-measurable activity but an increased soluble expression at higher temperatures in a heterologous E. coli host. PMID- 11421459 TI - Penaeus monodon (tiger shrimp) hemocyanin: subunit composition and thermostability. AB - Penaeus monodon (class Crustacea, order Decapoda) is one of the largest shrimps of the Penaeidea family from the Indo-West Pacific region. The dioxygen transporting protein hemocyanin, isolated from the hemolymph of this invertebrate, is composed of three 75-76 kDa structural/functional subunits designated as Pm1, Pm2 and Pm3. The N-terminal sequences of the chains were determined and compared with those of other decapodan hemocyanin subunits. Pm2 and Pm3 are highly homologous and electrophoretically undistinguishable polypeptides. In comparison to Pml, they have an extension of six residues. Pm1 is closely related to the subunit Pv2 of the Penaeus vannamei hemocyanin. Probably, subunits like Pm1 and Pv2 are family-specific for the Penaeidea hemocyanins and the other subunits are species-specific. Comparison of N-terminal sequences of respiratory proteins from the sub-orders Natantia and Reptantia demonstrated family- and sub-order-specific sequences. A melting point of 69 degrees C, lower than those for the di-hexameric decapodan hemocyanins, was determined from the temperature dependence of ellipticity of the mono-hexameric Penaeus monodon hemocyanin. Thermostability of decapodan hemocyanins depends on their aggregation state. PMID- 11421460 TI - Sex pheromone of the carpenterworm, Holcocerus insularis (Lepidoptera, Cossidae). AB - By means of thin-layer chromatography (TLC), electroantennogram (EAG), gas chromatography (GC), gas chromatography-mass spectrometry (GC-MS) and field tests, (Z)-3-tetradecenyl acetate(Z3-14:Ac), (E)-3-tetradecen-1-ol(E3-14:OH), and (Z)-3-tetradecen-1-ol(Z3-14:OH) at a ratio of 51:39:10 were identified from the female sex pheromone gland extracts of the carpenterworm, Holcocerus insularis Staudinger (Lepidoptera, Cossidae). The average amounts of Z3-14:Ac, E3-14:OH and Z3-14:OH in a single sex pheromone gland of calling moth were 7.29+/-2.72 ng, 5.72+/-2.43 ng and 1.44+/-0.56 ng, respectively. This is the first time that Z3 14:Ac was identified as a component of lepidopteran sex pheromone. Traps baited with rubber septa impregnated with Z3-14:Ac (500 microg/septum) were more effective than the traps baited with virgin female. The addition of the E3-14:OH and Z3-14:OH to rubber septa baited with Z3-14:Ac did not modify H. insularius male attraction, but E3-14:Ac slightly enhanced trap catch. PMID- 11421461 TI - Composition of the labial gland secretion of the bumblebee males Bombus pomorum. AB - Labial gland secretions of 22 males of the bumblebee Bombus pomorum, collected in the Czech Republic, were analysed separately for each individual. The secretions contained 70 compounds among which saturated and unsaturated hydrocarbons strongly dominated. The proportion of hydrocarbons in the secretion was unusually high (85-100%) compared to other bumblebee species studied so far (3-15%). Methyl and ethyl esters of fatty acids, known from many other bumblebee species, formed only minor components (less than 1% in sum) of the secretions of several B. pomorum individuals. No terpenic compounds, typical for males' marking secretion of many bumblebee species, were detected in B. pomorum. The absolute quantities of hydrocarbons present in the labial gland extracts were comparable with those usually present in other species. The composition of hydrocarbons found in the labial glands was different from the profile of the cuticular hydrocarbons. Despite our expectations in species exhibiting a regular patrolling and scent marking behaviour, the labial gland extracts obtained from B. pomorum males were unusually low concentrated and their chemical composition was atypical with respect of the proportions of hydrocarbons when compared with other patrolling species. This is the first report on the analysis of the labial gland secretion of the B. pomorum males. PMID- 11421462 TI - Effects of seasonality and fasting on the body mass and plasma growth hormone concentrations of the raccoon dog (Nyctereutes procyonoides) and the blue fox (Alopex lagopus). AB - Growth hormone (GH) promotes growth and endochondral ossification, but it is also important in the response to fasting due to its effects of increasing gluconeogenesis and lipolysis. In this study eleven raccoon dogs and blue foxes were followed for six months and their body mass and GH levels were measured. In November half of the animals of both species were put to a three-week fast. There were no significant differences in the GH levels between the animals of different ages and the subadults and adults both had quite low GH levels in the summer. Fasting had no effect on the GH levels of the raccoon dogs, but the fasting blue foxes had lower GH concentrations than the controls in Nov 16th. The control blue foxes experienced a significant increase in the GH levels in early November and the fasting blue foxes in late November. The GH concentrations of all the raccoon dogs rose in early December. As fasting did not cause an elevation in the GH levels but the concentrations increased with decreasing temperature and shortening daylength, the autumnal GH secretion of these species could be regulated by endogenous seasonal rhythms entrained by exogenous Zeitgebers such as temperature or photoperiod. The autumnal increase of GH levels contributes to the response to fasting as an adaptation to survive the winter months with inadequate nutrition. The raccoon dog which spends the coldest part of the winter in winter sleep seems to be better adapted to a total fast than the actively wintering blue fox. PMID- 11421463 TI - Tannins and related compounds: killing of amastigotes of Leishmania donovani and release of nitric oxide and tumour necrosis factor alpha in macrophages in vitro. AB - The antileishmanial and immunomodulatory potencies of a series of 28 polyphenols were evaluated in terms of extra- and intracellular leishmanicidal activity and macrophage activation for release of nitric oxide (NO), tumour necrosis factor (TNF) and interferon (IFN)-like properties. For this, several functional bioassays were employed including an in vitro model for leishmaniasis in which murine bone marrow-derived macrophages (BMMphi) were infected with the obligate intracellular parasite Leishmania donovani, an extracellular Leishmania proliferation assay, a fibroblast-lysis assay (TNF-activity), and a biochemical assay for NO. Except for gallic acid, its methyl ester, shikimic acid and catechin (EC50 25.8-67.9 nM) all polyphenols tested significantly inhibited the intracellular survival of L. donovani amastigotes (EC50 0.4-13.9 nM) when compared with the clinically used agent, sodium stibogluconate (EC50 10.6 nM). In contrast, none of the samples proved to be directly toxic for the extracellular promastigote form of the parasite. Noteworthy, the phenolic samples showed only moderate or no cytotoxicity against the murine host cells (EC50 10 to >144 nM). Although NO is an important effector molecule in macrophage microbicidal activity, the inducing potential of the test compounds for its release was found to be very moderate ranging from 7-54 microM (IFN-gamma/LPS 119 microM). On the other hand, inhibition of NO production had no apparent effect on intracellular leishmanicidal activity of polyphenols. Their in vitro TNF-inducing potential producing 50% lysis in murine L929 cells increased in the order of simple phenols and flavanols (34-48 U/ml) < A-type proanthocyanidins (53-80 U/ml) < B-type proanthocyanidins (64-200 U/ml) < hydrolyzable tannins (287-350 U/ml) at the host cell subtoxic concentration of 50 microg/ml. Furthermore, gallic acid and some hydrolyzable tannins showed appreciable IFN-like activities (14-23 U/ml) as reflected by inhibition of the cytopathic effect of encephalomyocarditis virus on fibroblast L 929 cells. The results provide a rational basis for the recorded anti-infectious efficacy of traditionally used herbal medicines containing tannins in vivo, in the light of both only moderate direct antimicrobial activities of distinct polyphenols in vitro and the limited knowledge on their uptake in humans. PMID- 11421464 TI - Constant expression of cyclooxygenase-2 gene in prostate and the lower urinary tract of estrogen-treated male rats. AB - Expression of cyclooxygenase-2 (E. C. 1.14.99.1) in prostate and the lower urinary tract (LUT) of the neonatally estrogenized male rat has been studied by using a COX-2's PCR fragment of 724 nt spanning 3 introns and a 478nt internal standard for quantitative RT-PCR. The same fragment of 724 nt was used for RNA probe in Northern hybridization. Neonatal estrogenization (10 microg/day of diethylstilbestrol on days 1-5) had no effect on COX-2 expression in prostatic urethra, prostatic lobes, or bladder. Acute estrogen treatment of castrated animals did not induce COX-2 expression, either. In addition the differential expression of basal level of COX-2 in the different lobes of prostate in normal rat was demonstrated. Our results suggest a constant expression of COX-2 gene in prostate and the lower urinary tract of the neonatally estrogenized (neoDES) rats. The present study indicates that the increased expression of COX-2 is probably not essential for the estrogen-driven development of stromal inflammation or hyperplastic and dysplastic alterations in the prostate of neoDES rats. PMID- 11421465 TI - Self-organization of local cortical circuits and cortical orientation maps: a nonlinear Hebbian model of the visual cortex with adaptive lateral couplings. AB - A nonlinear, recurrent neural network model of the visual cortex is presented. Orientation maps emerge from adaptable afferent as well as plastic local intracortical circuits driven by random input stimuli. Lateral coupling structures self-organize into DOG profiles under the influence of pronounced emerging cortical activity blobs. The model's simplified architecture and features are modeled to largely mimik neurobiological findings. PMID- 11421466 TI - Nuclei of plants as a sink for flavanols. AB - Onion cepa (L.) and Tsuga canadensis (L.) Carr. were investigated histochemically on the association of flavanols to nuclei. The young roots of Onion cepa are totally devoid of flavanol structures. Therefore, the excised roots tips were directly incubated into different solutions of flavanols. After 3 h of incubation a flavanol binding on the nuclei was recognizable, as seen by a yellowish-brown tanning reaction. Still to ensure the presence of flavanols on the nuclei, subsequent staining with the p-dimethylaminocinnamaldehyde reagent (DMACA) resulted in an intense blue colouration. Tsuga canadensis has significant amounts of vacuolar flavanol deposits in all parts of the tree as indicated by the DMACA reagent. It is obvious that also the nuclei were associated strongly with flavanols which can be demonstrated particularly elegant in the cells of the seed wings by histochemical methods. However, the mode of flavanol release from the original deposits is not yet clear. PMID- 11421467 TI - Familial related risk-factors in the development of chronic bronchitis/emphysema as compared to asthma assessed in a postal survey. AB - There is a lack of knowledge to which extent heredity or familial risk factors are involved in the development of chronic bronchitis/emphysema (CBE). Smoking is regarded as the most important risk factor, but only about 15% of smokers develop airway obstruction. We evaluated the importance of familial risk factors compared to smoking and ex-smoking using an epidemiological approach. In 1992, a postal questionnaire was distributed to a study sample. In all, 43 questions were asked, in a previously evaluated questionnaire, regarding respiratory symptoms, self reported lung diseases, smoking habits and familial occurrence of chronic bronchitis and asthma. The questionnaire was sent to 12,073 adults living in the southernmost part of Sweden. The age range was 20-59 years with an equal gender distribution. The study sample was drawn from the population records. The questionnaire was answered by 8469 subjects (70.1%), of whom 392 subjects (4.6%) stated that they had or had had CBE and 469 subjects (5.5%) stated that they had or had had asthma. In a model with logistic regression using the five explanatory variables gender, age, familial occurrence for asthma, familial occurrence for CBE and current or ex-smoking the most important risk factors for CBE were familial occurrence for chronic bronchitis [Odds ratios (OR): 5.19, 95% confidence interval (CI): 4.09-6.60, p = 0.000] and current or ex-smoking (OR: 1.74, 95% CI: 1.41-2.14, p = 0.000). The most important risk factors for asthma were familial occurrence for asthma (OR: 3.71, 95% CI: 3.06-4.51, p = 0.000) and current or ex-smoking (OR: 1.33, 95% CI: 1.09-1.61, p = 0.004). We have found that familial occurrence for CBE in first degree relatives together with smoking is a stronger risk factor for the development of CBE than is smoking. PMID- 11421468 TI - Plasma retinol and tocopherol levels in greek elderly population from an urban and a rural area: associations with the dietary habits. AB - Data on plasma concentrations of retinol and alpha-tocopherol in elderly (over 65 years) living in the Mediterranean countries are sparse. In the current study, plasma retinol and alpha-tocopherol concentrations were determined in 200 healthy subjects (82 men and 112 women) residing in an urban and in a rural area in South Western Greece. High prevalence of low plasma retinol concentration was observed for both sexes in each area. The 16.7% of males in urban and 25.4% in rural area had plasma retinol levels below 0.3 mg/l. The corresponding values for female subjects were 26% and 20.3%. Plasma alpha-tocopherol was found to be within normal range in the rural area (only one male had plasma alpha-tocopherol below 5 mg/l) while in the urban area 68.3% of men and 62% of women had plasma alpha tocopherol values below 5 mg/l. Differences in dietary habits between subjects living in the rural and in the urban area were revealed through a weekly food frequency questionnaire. This could partly explain the difference in alpha tocopherol concentrations between the two areas. The results of this study revealed high risk of vitamin A and E deficiency and suggests that the health care system in Greece should pay more attention on the social and health status of its elderly population. PMID- 11421469 TI - Prediction of mortality for congestive heart failure patients: results from different wards of an Italian teaching hospital. AB - Congestive heart failure (CHF) constitutes an important public health problem in Italy, evidenced by the high number of hospital admissions each year. Significant inter-hospital as well as interward differences in mortality rates for CHF patients that have been described may, in part, be explained by the differences in the severity of the illness of admitted patients. The goal of this study was to predict 30-day severity-adjusted mortality risk in patients with CHF admitted to wards of a teaching hospital in Siena, Italy, in 1997. A 30-day mortality was determined by linking hospital discharge files with the Tuscany Mortality Registry database. The 3M all patient refined diagnosis related group (APR-DRG) software was used as a risk assessment method. The relationships between death and the following variables were studied by univariate analyses: APR-severity risk, APR-mortality risk, age, sex, length of stay and, discharge ward. Multivariate analysis was also performed to verify the associations between death and those parameters found to be significant by univariate analysis. Unadjusted mortality proportions ranged from 4.3 to 44.0%. Logistic regression analysis demonstrated that APR-mortality risk, length of stay, and discharge ward were significantly and independently associated with 30-day mortality risk in patients with CHF. In summary, 30-day mortality risk varied significantly according to the ward of discharge in an Italian teaching hospital, even after adjustment for severity of illness. PMID- 11421470 TI - Environmental and serological evidence for the presence of toxocariasis in the urban area of Ancona, Italy. AB - To evaluate the Toxocara spp. eggs environmental contamination of the soil of the urban or suburban area Ancona (Italy), 22 public playgrounds were selected and several cores of soil were taken from any selected areas. To study the Toxocara seroprevalence in the inhabitants of Ancona, blood samples were collected from selected groups of individuals. One hundred and sixty-three blood samples were tested using an enzyme linked immunosorbent assay (ELISA) technique (Lofarma Lab, Milan, Italy) for the detection of IgG-specific antibodies to T. canis excretory secretory antigens. Toxocara spp. eggs were found in the soil samples from 14 (63.6%) playgrounds. Toxocara seroprevalence was detected in blood specimens from six (8.1%) out of 74 symptomatic individuals and from one (1.1%) out of 89 asymptomatic controls. Among symptomatic individuals, the association between Toxocara seroprevalence and eosinophilia resulted statistically significant (p = 0.029). The high environmental contamination frequency found make necessary to prompt preventive public health measures, such as control of stray animals, treatment of infected pets and hygiene education of the population. PMID- 11421471 TI - The relationship between urban airborne pollution and short-term mortality: quantitative and qualitative aspects. AB - The influence of airborne pollution on mortality rates has been examined since the well known severe pollution episodes of Pennsylvania (1948) and London (1952). Three main epidemiological approaches are available: transversal studies, time series studies, prospective studies. The most frequently used method is the time series analysis, which retrospectively determines daily mortality rates as compared to daily pollution levels in a defined period. The vast majority of studies applying this approach confirm the existence of a positive correlation between mortality and airborne pollution. The relative risk of death during the most severely polluted days as compared to the least polluted days ranges between 1.02 and 1.13. Airborne pollution affects mortality as related to cardiovascular and chronic obstructive pulmonary diseases mainly in subjects previously affected by these pathologies. The dose-response mortality-pollution curve shows linear increments of mortality at low pollution levels, weak increments at high pollution levels. This phenomenon is attributed to the existence of subjects highly susceptible to pollution induced adverse health effects. These subjects, dying at low pollution levels, are not available to increase mortality rates at high pollution levels. Mortality is mainly related to the concentration of single pollutants, such as suspended particle matter, sulfur oxides, nitric oxides and ozone. Elderly are highly susceptible to pollution-induced mortality increases. Therefore, the study of mortality-pollution correlation is of particular interest in cities characterized by a relative prevalence of the elderly in the population. The latency period between pollution increases and related mortality increases falls in the 1-3 days interval range. The pollution-mortality relationship is influenced by many factors, such as the occurrence of exceedingly high or low environmental temperatures, influenza epidemics, etc. Many hypotheses has been raised to support the causality of the mortality-pollution association, including a direct effect of pollutants on cardiovascular and respiratory apparatus, and the release of inflammatory mediators affecting blood viscosity and pneumocytes homeostasis. On the whole, available data suggest that pollution is able to increase mortality only if associated to other risk factors determining an increased susceptibility in the exposed population. PMID- 11421472 TI - Enterovirus surveillance of Italian healthy children. AB - Surveillance of acute flaccid paralysis (AFP) is the golden strategy recommended by the WHO to verify the condition of polio eradication in a country. Because of the difficulty to detect all of the expected AFP cases and to reach the target incidence of 1/100,000 requested by WHO, the surveillance of enteroviruses in the population has been adopted by several countries as an important additional method to verify the absence of wild-poliovirus circulation. To complete the results of AFP surveillance set up in Italy in 1996, we have conducted a wild poliovirus surveillance by examining stool samples from 1551 healthy children aged less than 5 years, collected during the period January 1997 to January 1998. The children were from three cities (Parma, Rome and Bari) located in northern, central and southern Italy. Thirty-nine polioviruses, 72 non-polio enteroviruses and 50 enteric, nonentero (NE), viruses were isolated from stool specimens. Polioviruses identified were nine type 1, seven type 2 and twenty-three type 3. Characterization of isolates by both antigenic and molecular methods showed that all polioviruses were of vaccine origin. As expected, most polioviruses, especially types 2 and 3, presented retromutations known to be associated with loss of the Sabin attenuated phenotype. The results of this study support the data obtained from the active AFP surveillance conducted in Italy in the same period--on the absence of paralytic disease due to wild poliovirus--and altogether demonstrate the effectiveness of the vaccination program. PMID- 11421473 TI - Immunity to diphtheria in Izmir, Turkey. AB - In order to assess immunity to diphtheria in Izmir, Turkey, a total of 743 persons 1-70 years of age were selected with cluster sampling. The information on socio-demographic characteristics, vaccination status and diphtheria history was gathered for each participant. Diphtheria antitoxin levels were measured qualitatively by using micro-enzyme immune assay. Of studied population, 79.1% had fully protective antitoxin levels (> or = 0.1 IU/ml). Diphtheria protection rates showed a gradual age-related decrease, reaching minimum in the 30-44 age group, in which 40.2% of these subjects had antibody titre below the full protective level. The diphtheria antitoxin geometric mean titer was highest in the 5-9 year age group (1.05 IU/ml). Then, geometric mean titer decreased with increasing age, and reached the minimum level in the 30-44 age group (0.19 IU/ml). These results suggest that in Izmir, Turkey, full serological protection against diphtheria is only detectable in 60% of the adult population. The enhancement of diphtheria immunity by booster vaccinations in adolescents and adults should be considered in Turkey. PMID- 11421474 TI - Prevalence and incidence of bloodborne viral infections among Danish prisoners. AB - In order to determine the prevalence and incidence of bloodborne viral infections among prisoners, we conducted a prospective study in a Danish medium security prison for males. The prisoners were offered an interview and blood test for hepatitis and human immunodeficiency virus HIV at inclusion as well as at release from prison or end of study. Of 403 prisoners available 325 (79%) participated in the initial survey and for 142 (44%) a follow-up test was available. 43% (140/325) of the participants were injecting drug users (IDUs) of whom 64% were positive for hepatitis B (HBV) and 87% for hepatitis C (HCV) markers. No cases of HIV or human T lymphotropic virus (HTLV) were found. 32% of all prisoners could transmit HBV and/or HCV by blood contact. 70% of IDUs had shared injecting equipment, and 60% had injected inside prison. Only 2% of IDUs were vaccinated against HBV. Duration of injecting drug use, numbers of imprisonments, and injecting in prison were independently and positively associated with the presence of HBV antibodies among IDUs by logistic regression analysis. The HBV incidence was 16/100 PY (95% CI: 2-56/100 PY) and the HCV incidence 25/100 PY (1 140) among injecting drug users (IDUs). We conclude that IDUs in prison have an incidence of hepatitis B and C 100 times higher than reported in the general Danish population. They should be vaccinated against hepatitis B and new initiatives to stop sharing of injecting equipment in and outside prison is urgently needed. PMID- 11421475 TI - Epidemiology and prognostic factors in meningococcal disease in a small island population: Malta 1994-1998. AB - STUDY OBJECTIVE: To review the epidemiology of meningococcal disease in Malta over the period 1994-1998, and to identify factors at presentation and in the management of meningococcal disease which may influence mortality. DESIGN: All admissions with meningococcal disease to a national hospital in a population based study over the period 1994-1998 were studied retrospectively. MAIN RESULTS: Fifty-six cases were diagnosed over 1994-1998, the incidence rising from 0.8/100,000 to 7.2/100,000 total population (p < 0.0001). The median time interval from arrival at hospital to administration of parenteral antibiotic decreased over the 5-year period from 4.4 to 1.2 hours (p = 0.025), with no significant change in the case-fatality rate. There was no association between the time interval from arrival at hospital to parenteral antibiotic administration, and mortality. The following features at presentation were associated with increased mortality: older age (p = 0.03), meningococcaemia compared with meningitis (p = 0.05), shock (p < 0.0001), disseminated intravascular coagulation (p = 0.0001), a normal/low white blood cell count (p = 0.0003), a low platelet count (p = 0.0001) and a high serum creatinine (p = 0.003). CONCLUSIONS: The upsurge of cases in the population was accompanied by a decrease in intervention time in the general hospital, probably due to increased awareness of the disease. This study did not show a positive relationship between early in-hospital administration of antibiotics and improved survival, probably because antibiotics were given earlier to those with fulminant disease and, with therefore, an inherently worse outcome. Stratification of cases by severity on admission is recommended in future studies. PMID- 11421476 TI - Surveillance of wild polioviruses in patients with acute flaccid paralysis in Malta during 1998 and 1999. AB - Certification of global poliomyelitis eradication can only be accomplished when all countries have been certified as having achieved wild poliovirus eradication. In order to achieve certification of eradication of poliomyelitis in Malta an active acute flaccid paralysis (AFP) surveillance system comprising the whole population was set up. Surveillance became effective in January 1998. During 1998, there were nine reported cases of AFP (non-polio AFP rate: 2.38 per 100,000 population) two of whom were children under 15 years of age (non-polio AFP rate: 2.52 per 100,000 for the population aged <15 years). In 1999 five cases of AFP were reported (non-polio AFP rate: 1.32 per 100,000 population). One case occurred in a child under 15 years (non-polio AFP rate: 1.28 per 100,000 for population aged <15 years). Virological investigation did not detect any wild or Sabin-like polioviruses. PMID- 11421477 TI - Variation in survival in coastal and continental regions of Croatia--results of a longitudinal study. AB - After 25 years of follow-up, regional variations in survival were analysed on a sample of 3343 participants (1780 female, 1563 male) from three urban and three rural municipalities in Croatia. Age of participants was in the range 35-54 years at the beginning of the study (1969). Cox regression for general mortality singled out one continental rural municipality (Virovitica) with the lowest survival in both genders. The relationship between the risk of death and age at the beginning of the study was approximately linear throughout its range for men, and after the age of 45 for women. Men showed a trend of better survival in the coastal region, which was consistent with findings of a similar concurrent study of rural Croatian population. PMID- 11421478 TI - Seroprevalence of hepatitis A, B and C markers in Greek warship personnel. AB - A seroepidemiological study was conducted to assess the seroprevalence of hepatitis A, B and C markers in 285 males (mean age: 24.4+/-4.4 years) aboard a Greek warship. Two hundred and sixty three serum samples were tested. None was found to be positive for HAV antibodies, three persons (1.1%) were positive for HBsAg, four persons (1.5%) were positive for anti-HBc and one person (0.4%) was positive for anti-HCV. Forty-five persons (17.1%) had developed titles anti-HBs > 10 IU/L. The establishment of a vaccination policy against hepatitis A among warship personnel is strongly recommended. PMID- 11421479 TI - Quality of the information contained in the minimum basic data set: results from an evaluation in eight hospitals. AB - To assess the quality of the information included in the minimum basic data set (MBDS) of the eight public hospitals of the Murcia region in order to ascertain what should be improved to be valid and reliable. An external encoder performed a recoding of a random sample of hospital discharges, using the patients hospital records and comparing afterwards the information obtained with the one reflected in the MBDS databases. Quality was assessed using 12 criteria. The reviewed discharges sample consisted at least of 96 cases per hospital (Type I error = 0.05, Type II = 0.10, for the most unfavorable case). A total of 796 cases were reviewed. The MBDS disagreement percentages with the patient record data were higher for the clinical data, with 41.6% for the main diagnosis and 33.5% for the main surgical procedure, being in both cases higher in those hospitals that had used to codify just the discharge record with regard to those that did so with the complete patient record. The variation rate in the diagnosis-related group (DRG) assignment was of 29.6%, and there was a decrease in the case-mix index of 1.07397 when reviewing with the patient record to 1.05555 in the MBDS. Within the administrative data, the highest disagreement rate was for the physician that signs the discharge (60.5%) and the patient's address (31.6%). In many of these assessed aspects there are significant differences between hospitals. A reliability problem was identified in the collected data, which mainly affects the clinical variables. It is therefore advisable to carefully assess the use of this information (both the MBDS directly as well as its grouping through the use of patient classification systems), and the indicators derived from it as its quality is not guaranteed. Systematic assessment and quality control of the MBDS production is advised. PMID- 11421480 TI - Investigation of accuracy of death certificate completion and implications on mortality statistics in Greece. PMID- 11421481 TI - Epidemiologic characteristics of kidney malformations. AB - Infants with kidney agenesis or dysgenesis, infants with cystic kidneys, and infants with horseshoe kidneys were studied, based on data from three large and population-based congenital malformation registers: a total of 2666 infants among 5.83 million births. There is a strong variability between programs of the rates of registered unilateral kidney malformations and these are strongly over represented in dead infants or infants with other malformations. There is a male excess but this varies in strength between different types of kidney malformations and between bilateral and unilateral forms. An increased twinning rate was found. The different types of kidney malformation differed with respect to kind of associated non-urological malformations in multimalformed infants. We conclude that for monitoring purposes one should restrict analysis to bilateral forms but that unilateral forms can be of interest in epidemiological analyses. PMID- 11421482 TI - Assessment of the relationship of chronic opisthorchiasis to Epstein-Barr virus infection as well as some cytogenetical and immunological parameters in two comparable Siberian regions. AB - Previous studies have shown an increase in the frequency of chromosomal abnormalities in the peripheral blood lymphocytes from opisthorchiasis patients. Some evidence suggests that there is an association between chronic opisthorchiasis and certain herpes viruses. To study the relationship of opisthorchiasis to the reactivation of Epstein-Barr virus (EBV) infection as well as the influence of opisthorchis infection on some cytogenetical and immunological parameters, we used the indirect immunofluorescence for measuring some virus specific antibodies, the cytokinesis-block lymphocyte micronucleus assay, and the quantitative immunodiffusion method for measuring immunoglobulin concentrations in serum. A total of 1,580 people were monitored in two comparable Siberian regions: in the Ob River region which is endemically related to opisthorchiasis caused by Opisthorchisfelineus and in the nonendemic control Yenisey River region. There was no significant difference in each of the tested parameters between two uninfected controls from the endemic Ob and nonendemic Yenisey regions. We have found significant difference (p < 0.01) in the frequency of micronucleated cytokinesis-block lymphocytes and the antibody levels against certain EBV antigens between the examined inhabitants of the opisthorchis infected Ob and uninfected Yenisey regions. Furthermore, there were a good correlation (r = + 0.72) between the increase in titres of antibody to the EBV capsid antigen and the high frequency of micronucleated lymphocytes in the opisthorchis-infected Ob population. Also, levels of both IgG and IgM were increased in opisthorchiasis patients. This study confirms an association between chronic opisthrochiasis and reactivation of EBV which may be implicated in the development of cancer in opisthorchiasis patients. PMID- 11421483 TI - Systemic proliferative changes and clinical signs in cynomolgus monkeys administered a recombinant derivative of human epidermal growth factor. AB - Epidermal growth factor (EGF) effects have been explored extensively in vivo in rodents, but little is known about trophic responses in nonhuman primates. A previous publication reports the hyperplastic epithelial/parenchymal changes noted in the digestive tract (tongue, esophagus, stomach, intestine, liver, gallbladder, pancreas, and salivary glands) of adult cynomolgus monkeys treated with recombinant human EGF(1-48) (rhEGF(1-48)). This report documents clinical findings and structural effects in the remaining epithelium-containing tissues of these animals. Two monkeys/sex/dose received rhEGF(1-48) by intravenous bolus at 0 (vehicle), 10, 100, 500 (females only), or 1,000 microg/kg/day (males only) daily for up to 2 weeks. Treatment- and dose-related clinical findings included emesis, fecal alterations (soft feces and diarrhea), lacrimation, nasal discharge, hypoactivity, transient hypotension, and salivation after dosing. Male monkeys administered 1,000 microg/kg became moribund after 5 days of treatment and were necropsied. All other monkeys completed the 2-week treatment period. Necropsy findings in nongastrointestinal tissues were: enlarged, pale kidneys at 100 microg/kg and greater; small thymuses seen sporadically at all doses; and enlarged adrenals and small thyroids in males at 1,000 microqg/kg. Respective organ-to-brain weight ratios at 500 and 1,000 microg/kg for kidneys were 1.5- and 2.6-fold greater and for heart were 1.7- and 1.3-fold greater than controls. Microscopically, pronounced dose-related epithelial hypertrophy and hyperplasia were evident in kidney, urinary bladder, skin (epidermis and adnexa), mammary gland, prostate, seminal vesicles, epididymis, uterus, cervix, vagina, thyroid, thymus, tonsillar crypts, cornea, trachea, and pulmonary airways. Epitheliotrophic effects were conspicuous in many tissues at 100 to 1,000 microg/kg. Changes to renal collecting ducts were present at 10 microg/kg, suggesting that kidneys were a relatively sensitive target. Proliferative alterations were not apparent in testes, intraocular structures, brain ependyma and choroid plexus at any dose. Aside from the noted exceptions, rhEGF(1-48) was a pantrophic epithelial mitogen in cynomolgus monkeys when used intravenously at suprapharmacologic doses. PMID- 11421484 TI - Toxicologic lesions associated with two related inhibitors of oxidosqualene cyclase in the dog and mouse. AB - Two novel hypolipidaemic agents, both members of the aminopyrimidine series, with a mode of action of inhibition of oxidosqualene cyclase (OSC), were administered orally to dogs and mice for 14 and 28 days. Both compounds produced a similar spectrum of pathologic changes. In dogs, the agents produced equatorial single cell necrosis and cataract in the lens (also observed clinically); atrophy, ulceration, and inflammation of the cornea; hyperkeratosis, acanthosis, hair papillary atrophy, and inflammation of the skin; and epithelial degeneration and sperm granuloma in the epididymides. One female dog showed signs of liver toxicity. In mice, severe cataract formation was seen with both compounds, and liver toxicity was produced by one of the compounds. The severity and speed of onset of the cataract formation were very marked. The changes seen were dissimilar to those reported with the most commonly used class of hypolipidaemic agents in the clinic, the hydroxymethyl glutaryl coenzyme A (HMGCoA) reductase inhibitors but were reminiscent of those reported for the hypolipidaemic agent Triparanol. which was predictive of toxicity seen in man. PMID- 11421485 TI - Dose-dependent induction of glandular stomach preneoplastic and neoplastic lesions in male F344 rats treated with catechol chronically. AB - The dose-dependence of catechol glandular stomach carcinogenesis was investigated in male F344 rats. Groups of 30 male animals were fed catechol at dietary levels of 0 (control). 0.1, 0.2, 0.4, and 0.8% for up to 104 weeks. Five rats of each group were killed at 34 weeks and the remaining animals were sacrificed at the termination, all undergoing histopathological examination. Moderate retardation of body weight increase was observed in the 0.8% group. but no adverse effects were found in terms of survival. Submucosal hyperplasias and adenomas of the pyloric glands developed in the 0.4 and 0.8% groups, only very minor changes being noted in the 0.1 and 0.2% groups at week 34. Incidences of adenocarcinoma development in the pylorus were 4% and 8% in 0.4% and 0.8% groups, respectively, and 0 in the 0.1% and 0.2% groups, at the termination. Adenomas and submucosal hyperplasias were found in nearly all animals fed 0.2% catechol or more, the incidences of those in 0.1% group being 0% and 56%, respectively. Serum gastrin levels were significantly increased in the 0.2, 0.4, and 0.8% groups at 34 weeks, and in all treated groups at the termination, at extents comparable with the induction of proliferative lesions in the pylorus. The results thus demonstrated that dietary levels of 0.4% and 0.8% catechol long-term induce adenocarcinomas in the pyloric glands, while 0.1 and 0.2% cause benign proliferative lesions, all accompanied by increase in serum gastrin levels. As a no-effect level could not be decided in the present study, further investigation of lower doses is needed to determine whether a threshold exists. PMID- 11421486 TI - Pathology of ocular irritation with acetone, cyclohexanol, parafluoroaniline, and formaldehyde in the rabbit low-volume eye test. AB - The ocular irritation responses to 11 different surfactants and two concentrations of acetic acid and sodium hydroxide have been shown to depend on the extent of initial injury, despite marked differences in the processes leading to tissue damage. The purpose of these studies was to determine the extent to which this fundamental relationship applies to other nonsurfactants. Ten microl of acetone (ACT). cyclohexanol (CY), parafluoroaniline (PF), or 37% formaldehyde (FA) was directly applied to the cornea of the right eye of each rabbit. Eyes and eyelids were macroscopically scored for signs of irritation beginning 3 hours after dosing and periodically until recovery or 35 days. Tissues were obtained for light microscopic examination after 3 hours and on days 1, 3, and 35. Initial corneal injury was characterized quantitatively at 3 hours and I day using in vivo confocal microscopy (CM) and by postmortem quantitation of dead corneal epithelial cells and keratocytes using a Live Dead Assay (L/D, Molecular Probes) and scanning laser CM. Corneal changes over time were characterized quantitatively using in vivo CM performed at 3 hours and 1, 3, 7, 14, and 35 days. The changes with ACT were consistent with mild irritation. Corneal injury was limited to the epithelium and superficial stroma, with the mean normalized depth of injury (NDI) being less than 10% with the majority of regions showing no stromal injury. Changes with CY and PF were consistent with moderate to severe irritation, and FA caused severe irritation. Specifically, corneal injury by CY and PF tended to involve the epithelium and anterior stroma, with the mean NDI being 10.4% to 23.8%, while injury with FA involved the epithelium, deep stroma, and at times the endothelium. Interestingly, with FA significantly less injury was observed at 3 hours with a dramatic increase in injury observed at 1 day and thereafter. In conclusion, these results continue to support and extend our hypothesis that ocular irritation is principally defined by the extent of initial injury despite clear differences in the means by which irritants cause tissue damage. We believe this approach can be applied to developing alternative assays based on injury to ex vivo eyes or injury to an in vitro corneal equivalent system. PMID- 11421487 TI - Suppressive effects on allergic contact dermatitis by short-term fasting. AB - Fasting alters various hormonal and immune conditions. It has been reported that delayed type immune response to the injection of keyhole limpet hemocyanin was depressed by short-term fasting. In this study, we adopted the computer-assisted image analyzer for histopathological analysis and evaluated the influence of short-term fasting on allergic contact dermatitis induced by 2,4 dinitrofluorobenzene (DNFB). Mice were sensitized by painting of DNFB to the abdomen. After the sensitization, mice were challenged by DNFB painting to the ear. Fasting started 24 hour before (48-hour fasted group) or immediately after (24-hour fasted group) the challenging. Fasting without DNFB treatment did not induce remarkable change of ear thickness, ear tissue, serum albumin, serum total protein, serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase. or serum creatine phosphokinase. In contrast, lasting suppressed the increment of ear thickness in the DNFB-treated group in this study. We could also demonstrate, using the computerized image analyzer, that both lymphocyte infiltration and the edema in the dermis were suppressed in fasted mice treated with DNFB. Further, edema in the dermis was inhibited more strongly in 48-hour fasted mice than in 24-hour lasted mice. These findings indicate that short-term fasting induce histopathological changes in the state of contact dermatitis. PMID- 11421488 TI - Glutamate receptors in peripheral tissues: current knowledge, future research, and implications for toxicology. AB - We illustrate the specific cellular distribution of different subtypes of glutamate receptors (GluRs) in peripheral neural and non-neural tissues. Some of the noteworthy locations are the heart, kidney, lungs, ovary, testis and endocrine cells. In these tissues the GluRs may be important in mediating cardiorespiratory, endocrine and reproductive functions which include hormone regulation, heart rhythm, blood pressure, circulation and reproduction. Since excitotoxicity of excitatory amino acids (EAAs) in the CNS is intimately associated with the GluRs, the toxic effects may be more generalized than initially assumed. Currently there is not enough evidence to suggest the reassessment of the regulated safety levels for these products in food since little is known on how these receptors work in each of these organs. More research is required to assess the extent that these receptors participate in normal functions and/or in the development of diseases and how they mediate the toxic effects of EAAs. Non-neural GluRs may be involved in normal cellular functions such as excitability and cell to cell communication. This is supported by the wide distribution in plants and animals from invertebrates to primates. The important tasks for the future will be to clarify the multiple biological roles of the GluRs in neural and non-neural tissues and identify the conditions under in which these are up- or down-regulated. Then this could provide new therapeutic strategies to target GluRs outside the CNS. PMID- 11421489 TI - Peroxisome proliferator-activated receptors in atherosclerosis and inflammation- an update. AB - Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear receptor subfamily of transcription factors with pleiotropic effects on intra- and extracellular lipid metabolism, glucose homeostasis, cell proliferation, control of inflammation, and atherosclerosis. Three PPARs, namely alpha, delta, and gamma have been identified with distinct tissue distribution patterns and metabolic functions. PPAR-alpha is predominantly expressed in brown adipose tissue, liver, kidney, duodenum, heart, skeletal muscle, and vascular endothelial cells and is involved in the control of lipoprotein metabolism, fatty acid oxidation, and the cellular uptake of fatty acids. PPAR-gamma is highly expressed in brown and white adipose tissues and, to lesser extent, in large intestine, retina, and some parts of the immune system, and plays a critical role in adipocyte differentiation and fat deposition. PPAR-delta shows a widespread tissue distribution but its regulation and functions are not yet known. Considerable evidence indicates that PPARs (PPAR-alpha and PPAR-gamma) have beneficial effects in inflammatory diseases, including atherosclerosis, through regulation of cytokine production, adhesion molecule expression on the endothelial cells, fibrinolysis, and modulation of monocyte-derived macrophages. In this review, the general and specific roles of the PPAR isotypes and their implications in the control of vascular inflammation and atherosclerosis are discussed. PMID- 11421490 TI - Specific genomic alterations in rat renal cell carcinomas induced by N-ethyl-N hydroxyethylnitrosamine. AB - To characterize genetic alterations occurring in renal tumorigenesis, EHEN induced renal cell tumors were examined using restriction landmark genomic scanning (RLGS) analysis, an electrophoretic separation technique that detects gene amplifications and deletions. Comparison of DNAs from tumor against those from corresponding nontumorous kidney and/or EHEN-treated kidney without development of renal tumors yielded specific alterations in terms of both amplified and reduced DNA spots. Two amplified spots were detected only in renal cell tumors and an additional four spots were frequent in EHEN-treated kidneys. One reduced spot was common to all tumor samples, and another was frequently detected in the tumors analyzed but not in EHEN-treated kidneys. A subset of the altered spots thus appeared to be specific for EHEN-induced renal tumorigenesis. PMID- 11421491 TI - Morphological and immunohistochemical characteristics of the heterogeneous prostate-like glands (paraurethral gland) seen in female Brown-Norway rats. AB - Morphological and immunohistochemical characteristics of the prostate-like glands (paraurethral gland) seen spontaneously in female Brown-Norway (BN) rats were investigated by gross, and light and electron microscopic examination. At 9- to 10-weeks-old, the paraurethral gland was detected in 50 out of 52 female animals examined (96.2%), and it was observed as single or paired structures located ventrolaterally in the urethra just caudal to neck of the urinary bladder. Microscopically, the glandular acini consisted of flat to cuboidal secretory epithelium surrounded by the smooth muscle. The glands displayed modest secreting activity, and a few secreting materials were observed in the acinar lumens. The main peripheral ducts were located in the urethral wall, and drained into the urethra on both sides. Ultrastructurally, abundant rough endoplasmic reticulum (RER), numerous mitochondria and lysosomes, and secretory granules in the apical portion of the epithelial cells were noted, and basal cells were also observed. These gland epithelial cells showed positive reactions when stained for androgen receptor (AR), prostate specific antigen (PSA), or prostate specific acid phosphatase (PSAP). The nature of this paraurethral gland resembled that of the prostate gland in male rats. Thus, the paraurethral glands seen in the females were considered homologous to the prostate gland in males. PMID- 11421492 TI - Protective effect of rhIL-11 in a murine model of acetaminophen-induced hepatotoxicity. AB - Acetaminophen intoxication results in hepatotoxicity associated with increased serum concentrations of hepatocellular leakage enzymes such as aspartate aminotransferase, lactate dehydrogenase, and alanine aminotransferase, centrilobular degeneration and necrosis, and activation of Kupffer cells. Recombinant human Interleukin-11 (rhIL-11) downregulates the production of proinflammatory mediators from activated macrophages and has direct effects on hepatocyte gene expression. Based on these biological activities of rhIL-11, the effect of pretreatment with rhIL-11 in a murine model of acetaminophen-induced hepatotoxicity was examined. Administration of 500 microg/kg acetaminophen to B6C3F1 mice resulted in progressive hepatotoxicity as demonstrated by elevated serum concentrations of hepatocellular leakage enzymes and TNFalpha and histopathology. Pretreatment with 250 or 500 microg/kg of subcutaneously administered rhIL-11 2 hours before acetaminophen administration reduced serum concentrations of hepatocellular leakage enzymes and TNFalpha by 40-50%. This was associated with a statistically significant decrease in mean severity score for centrilobular hemorrhage and necrosis from grade 3 to grade 2 for rhIL-11-treated animals compared to vehicle. These results indicate that treatment with rhIL-11 has a protective effect in a model of acetaminophen-induced liver damage. PMID- 11421493 TI - Subchronic sodium chlorate exposure in drinking water results in a concentration dependent increase in rat thyroid follicular cell hyperplasia. AB - Chlorine dioxide (ClO2) is an effective drinking water disinfectant, but sodium chlorate (NaClO3) has been identified as a potentially harmful disinfection by product. Studies were performed to describe the development of thyroid lesions in animals exposed to NaClO3 in the drinking water. Male and female F344 rats and B6C3F1 mice were exposed to 0, 0.125, 0.25, 0.5, 1.0, or 2.0 g/L NaClO3 for 21 days. Additional male F344 rats were exposed to 0, 0.001. 0.01. 0.1, 1.0. or 2.0 g/L NaClO3 for 90 days. Female F344 rats were exposed to 0, 0.5, 1.0, 2.0, 4.0, or 6.0 g/L of NaClO3 for 105 days. Thyroid tissues were processed by routine methods for light microscopic examination, and follicular cell hyperplasia was diagnosed using a novel method. Thyroid hormone levels were altered significantly after 4 and 21 days. NaClO, treatment induced a concentration-dependent increase in the incidence and severity of thyroid follicular cell hyperplasia. Male rats are more sensitive to the effects of NaClO3 treatment than females. Follicular cell hyperplasia was not present in male or female B6C3F1 mice. These data can be used to estimate the human health risk that would be associated with using ClO2, rather than chlorine, to disinfect drinking water. PMID- 11421494 TI - Pulmonary function in nonsmokers following exposure to sidestream cigarette smoke. AB - Ten healthy male and 10 healthy female, "never-smoking" subjects (ages 21-50) participated in a 5-day environmental room study to determine if an acute exposure to a high level of fresh diluted sidestream smoke (FDSS) would alter pulmonary function. On Monday, Tuesday, Thursday and Friday, the twenty subjects sat in an environmental room for 7.33 hours and were exposed to filtered and humidified air. On Wednesday, the twenty subjects were exposed in an environmental room for 7.33 hours to an average respirable suspended particle (RSP) concentration of 179 micrograms per m3 of FDSS generated by machine smoking Kentucky 1R4F reference cigarettes. This level of FDSS is 3.3 times the 95th percentile concentration of workplace environmental tobacco smoke exposure levels previously measured in the US. FVC and FEV1 decreased approximately 1.6% (p < 0.05) in both males and females after exposure. Similarly, PEF decreased approximately 1.3% (p < 0.03) following exposure. The observed decrease in pulmonary function was consistent with a "stress" related norepinephrine-induced alteration in blood flow leading to transient bronchoconstriction. Alternatively, a cholinergic reflex due to activation of bronchopulmonary C fibers may have also played a role in the transient bronchoconstriction. These small exposure-related decrements in pulmonary function were reversible. PMID- 11421495 TI - Draft recommendations on classification of rodent neoplasms for Peto analysis. AB - The Executive Committee of the Society of Toxicologic Pathology appointed an ad hoc working group to review the current use of the Peto model for statistical analysis of rodent carcinogenicity study data and to provide recommendations for pathologists regarding appropriate and consistent classification of neoplasms for analysis by the Peto model. In the Peto model, neoplasms that are detected as in life observations are classified as Mortality Independent and the incidences of these neoplasms are analyzed by the onset-rate method. For neoplasms that are not detected in-life in animals that come to necropsy before the end of the treatment period, the original Peto model requires pathologists to classify neoplasms as Fatal or Incidental (Non-fatal). Each Fatal neoplasm is modeled statistically as rapidly fatal regardless of how long the neoplasm has been present in the animal, and the date of death is used as a surrogate for date of onset of Fatal neoplasms. Fatal neoplasms are analyzed by the death-rate (life-table analysis) method, while Incidental neoplasms are analyzed by the prevalence method. In practice, most neoplasms that cause death are not rapidly fatal, e.g., a slowly progressive pituitary adenoma in a rat. The Peto model does not appropriately model slowly growing or slowly progressive neoplasms that cause death. RECOMMENDATIONS: 1. For animals that die or are killed prior to scheduled sacrifice, all neoplasms should be classified into one of four categories on an animal-by-animal basis: Observed In Life (Mortality-lndependent)--Neoplasms that were observed during in-life observations and for which the day of onset (first observation) was recorded: Incidental--Neoplasms that were not observed prior to necropsy and likely did not contribute to death of the animal or bringing the animal to necropsy; Rapidly Fatal--Neoplasms that were not observed prior to necropsy, likely contributed to death or bringing an animal to necropsy, and appeared to grow and develop rapidly. For these neoplasms, the day of death would serve as a surrogate for the day of onset of the neoplasm: Not Rapidly Fatal- Neoplasms that were not observed prior to necropsy, likely contributed to death or bringing an animal to necropsy, and did not appear to grow or develop rapidly. 2. Other guidelines for pathologists interpreting carcinogenicity studies include: More than one neoplasm may be Rapidly Fatal, Not Rapidly Fatal, and/or Incidental in a single animal. An individual animal may have 2 or more Rapidly Fatal neoplasms if the pathologist believes that each neoplasm had a rapid onset and likely contributed to bringing the animal to necropsy: Each pathologist should use scientific judgment when differentiating Rapidly Fatal and Not Rapidly Fatal neoplasms. This scientific judgment is similar to that used in determining whether neoplasms contribute to the cause of death. Criteria for malignancy (tissue invasion, metastasis, necrosis, high mitotic index, anaplasia, etc.) can be helpful as guides for classifying a neoplasm as Rapidly Fatal. 3. Industry and regulatory statisticians should continue to explore alternatives to the Peto method that do not require classification of neoplasms according to their impact on the death of the animal, and should use these methods if they offer equivalent or superior analytical capabilities. PMID- 11421496 TI - A survey of discriminant methods used in toxicological histopathology. AB - A cluster survey of United Kingdom (UK)-based toxicological pathologists has been made of the methods used to discriminate treatment-related from spontaneous variation in histopathological material from toxicity studies. The response rate was 78% of 102 polled. Respondents overwhelmingly preferred methods based on grading or scoring lesions and comparatively avoided methods based on ranking, measuring, counting, pairwise comparisons, or overviewing. This preference was in spite of regarding parametric and ranking methods as both more powerful and specific than some commonly used scoring methods. Respondents were evenly divided on the use of blind examination and tended to avoid using historic material to form an opinion. PMID- 11421497 TI - Diet modification alone did not improve the health and survival of rats. PMID- 11421498 TI - New hope for the treatment of multi-drug-resistant tuberculosis? PMID- 11421499 TI - A review of long-term oxygen therapy for chronic obstructive pulmonary disease. AB - This study aimed to review the evidence for the use of long-term oxygen therapy for patients with chronic obstructive pulmonary disease (COPD). The design was a systematic Cochrane review of randomized controlled trials (RCTs) of long-term oxygen therapy for COPD and main outcome measure was survival on home oxygen therapy. Five RCTs were identified. Data from two trials of nocturnal oxygen therapy in mild to moderate hypoxaemia were aggregated. Data from the other three trials could not be aggregated because of differences in trial design and patient selection. Treatment with continuous versus nocturnal oxygen therapy produced a significant improvement in mortality after 24 months [Peto odds ratio 0.45, 95% confidence interval (95% CI) 0.25-0.81] for the continuous therapy group. Treatment with oxygen therapy versus no oxygen therapy showed a significant improvement in mortality after five years in the group receiving oxygen therapy (Peto odds ratio 0.42, 95% CI 0.18-0.98). There was no difference in mortality for patients with COPD and mild to moderate daytime hypoxaemia and nocturnal desaturation receiving nocturnal oxygen therapy versus no oxygen therapy or sham treatment. Long-term oxygen therapy versus no oxygen therapy in patients with COPD and moderate hypoxaemia had no effect on survival. In conclusion, long-term oxygen therapy improved survival in a selected group of COPD patients with severe hypoxaemia but few co-morbidities. Long-term oxygen therapy did not improve survival in patients with moderate hypoxaemia or in those with mild to moderate hypoxaemia and arterial desaturation at night. PMID- 11421500 TI - Does immunotherapy with heat-killed Mycobacterium vaccae offer hope for the treatment of multi-drug-resistant pulmonary tuberculosis? AB - The ability of immunotherapy with heat-killed Mycobacterium vaccae (NCTC 11659), as an addition to the available chemotherapy, to improve the outcome in patients with multi-drug-resistant tubercle bacilli (MDRTB) who had not been cured by chemotherapy alone was evaluated in tuberculosis centres in Estonia, Iran, Kuwait, New Zealand, Romania, Vietnam and the U.K. A total of 337 patients in the above countries received intradermal injections of M. vaccae in addition to chemotherapy. Patients were grouped according to the length of their histories of disease: less than or greater than 2 years duration. Initially, single doses of M. vaccae were given but subsequently up to 12 doses at 2-month intervals were given. Chemotherapy varied from isoniazid alone to drugs selected according to susceptibility tests. Most patients had failed to respond to repeated courses of chemotherapy and the majority, were expected to die from their disease. Results were assessed by sputum smear and culture and by clinical observations. Cured patients were followed for 18-24 months to exclude relapse. Eighteen of 22 (82%) patients with disease for less than 2 years were bacteriologically cured by one or two doses of M. vaccae. Among 315 chronic patients, 24 (7.6%) were cured after one dose, 37.9% after seven doses and 41.6% after 12 doses. Sixty-six chronic patients were lost to follow-up, or died, during the multi-dose regimens. Nine of 33 patients (27%) with advanced disease unaffected by several courses of chemotherapy and discharged on isoniazid alone in Vietnam were cured by 3-12 injections of M. vaccae. The data provide preliminary evidence that the addition of immunotherapy with M. vaccae to chemotherapy improves the rate of cure of MDRTB, most effectively in patients with short histories of disease, but multiple dosing can have beneficial effects in chronic patients in whom chemotherapy has failed. A randomized clinical trial of this immunotherapy in MDRTB patients is therefore required. PMID- 11421501 TI - Long-term administration of N-acetylcysteine decreases hydrogen peroxide exhalation in subjects with chronic obstructive pulmonary disease. AB - Patients with chronic obstructive pulmonary disease (COPD) exhale more hydrogen peroxide (H2O2) and lipid peroxidation products than healthy subjects. This may reflect oxidative stress in the airways that plays important role in the development and progression of COPD. N-acetylcysteine (NAC), a mucolytic drug, possesses antioxidant properties as it is a precursor of reduced glutathione that together with glutathione peroxidase may decompose H2O2 and lipid peroxides. We aimed to determine the effect of NAC, 600 mg effervescent tablets (Fluimucil), once a day for 12 months, and placebo on the concentration of H2O2 and thiobarbituric acid reactive substances (TBARs) in expired breath condensate and serum levels of two lipid peroxidation products (TBARs, lipid peroxides) in patients with COPD. The study was performed as a double-blind, double-dummy comparison between active drug and placebo in two parallel groups. Forty-four outpatients with stable COPD (22 in the NAC group and 22 in the placebo group) completed the study. Specimens of expired breath condensate and serum were collected at the randomization visit and then every 3 months over 1 year. The concentration of TBARs and H2O2 in expired breath condensate was measured spectrofluorimetrically by the thiobarbituric acid and homovanillic acid methods, respectively. Serum levels of lipid peroxides were determined spectrophotometrically after extraction with butanol and pyridine. Initially, H2O2 exhalation did not differ between the placebo and NAC groups up to 6 months of treatment. After this the significant differences were observed. After 9 and 12 months of treatment NAC group exhaled 2.3-fold (0.17+/-0.33 microM vs. 041+/ 0.26 microM, P<0.04) [median 0.01 microM, quartile range (qr)=0.22 vs. median 0.15 microM, qr =0.43] and 2.6-fold (0.15+/-0.23 microM vs. 0.40+/-0.25 microN, P<0.05) median = 0.00 microM, qr = 0.23 vs. median = 0.36 microM, qr = 0.51] less H2O2 than placebo receivers, respectively. No significant effect of NAC administration on TBARs exhalation and serum levels of TBARs and lipid peroxides were noted over the whole treatment period. Also no significant associations between exhaled H2O2 and concentrations of lipid peroxidation products were noted in both treatment groups at any time-point. These results indicate that long-term oral administration of NAC attenuates H2O2 formation in the airways of COPD subjects and prove anti-oxidant action of drug. However, further studies are necessary to estimate the clinical significance of this finding. PMID- 11421502 TI - Relationship between airway sensitivity to adenosine 5' monophosphate and the shape of the concentration-response curve to methacholine in subjects with allergic rhinitis. AB - The objective of this study was to determine differences in airway sensitivity to adenosine 5'-monophosphate (AMP) between allergic rhinitis subjects with plateau and those without evidence of plateau on the concentration response curves to methacholine. A total of 51 adults (38 subjects with allergic rhinitis and 13 healthy controls) were challenged with increasing concentrations of methacholine and AMP. The methacholine challenge was terminated when there was a 40% or more decrease in forced expiratory volume in 1 sec (FEV1), whereas the AMP challenge was stopped when FEV1 had fallen by more than 20%. A plateau for methacholine was detected in all 13 healthy controls and in 27 patients with allergic rhinitis (AR plateau group), whereas 11 subjects with allergic rhinitis did not exhibit a plateau (AR-non-plateau group). The median (range) PC20 AMP (provocative concentration required to produce a 20% fall in FEV1) value for the AR-non plateau group was 44.0 mg ml(-1) (3.3-400.0), compared with 400.0 mgml(-1) (12.1 400.0) in the AR-plateau group (P=0.03) and 400.0 mgml(-1) in the healthy control group (P=0.007). The proportion of subjects who showed bronchoconstriction in response to AMP was higher in the AR-non-plateau group (73%) than in the AR plateau group (30%) (P=0.03). However, three subjects with allergic rhinitis who had normal sensitivity to methacholine and plateau showed bronchoconstriction in response to AMP. We conclude that, in subjects with allergic rhinitis, the absence of plateau on the concentration response curves to methacholine is associated with a higher prevalence and degree of bronchoconstriction in response to AMP. However, the two bronchoconstrictor stimuli were not identifying the same abnormalities of the airways. PMID- 11421503 TI - Airway responsiveness as a direct factor contributing to the dyspnoea perception in asthma. AB - It is not clear whether airway responsiveness is directly related to the perception of bronchoconstriction in asthma. The purpose of this study is to directly compare the perception of induced bronchoconstriction among the groups classified according to the degree of airway responsiveness. Two hundred and twenty-seven patients with the definitive or suspected asthma underwent a methacholine provocation test. During the test, the degree of dyspnoea was assessed by a modified Borg scale. The perception of induced bronchoconstriction was indicated by the slope in the linear regression analysis between changes in Borg score and the reduction in forced expiratory volume in 1 sec (FEV1) as a percentage of baseline value. The provocative concentration of methacholine resulting in 20% fall in FEV1 (PC20) was calculated. The degree of airway responsiveness to methacholine was categorized as moderate to severe airway hyper responsiveness (AHR) if PC20 was < 1 mgml(-1), mild AHR if PC20 was > or =1 but < or =4 mgml(-1), borderline AHR if PC20 was >4 but < or =16 mgml(-1), and normal airway responsiveness (negative AHR) if PC20 was > 16 mgml(-1). Positive AHR was defined as PC20< or =4 mgml(-1). Another index of bronchial responsiveness (BR index) was calculated as the log [(% decline in FEV1/log final methacholine concentration as mg dl(-1)+10]. We found that the geometric mean of the slope was lower in subjects with positive AHR (0.12, n=115) than in subjects with negative AHR (0.17, n=72; P<0.01). The geometric mean of the slope in subjects with borderline AHR (0.14, n=40) was between the two groups. Furthermore, the slope was decreased in asthmatics with moderate to severe AHR compared with mild AHR (P <0.05), although the baseline FEV1 did not differ between the two groups. In multiple regression analysis, airway responsiveness expressed as BR index had a significant effect on the perception of bronchoconstriction. We conclude that the perception of bronchoconstriction is diminished in patients with AHR and the degree of airway responsiveness may be directly related to the perception of bronchoconstriction in asthmatic subjects. PMID- 11421504 TI - Erythropoietic response to hypoxaemia in diffuse idiopathic pulmonary fibrosis, as opposed to chronic obstructive pulmonary disease. AB - The aim of this study was to assess the erythropoietic response to hypoxaemia in patients with diffuse idiopathic pulmonary fibrosis (DIPF), and to speculate on the underlying mechanisms. Patients on an established chronic respiratory failure due to DIPF or chronic obstructive pulmonary disease (COPD) were studied. The erythropoietic response to hypoxaemia in both conditions was assessed. We studied 18 patients with DIPF and 29 patients with COPD in respiratory failure in a stable stage, free from acute infection and congestive heart failure. Blood gases, erythrocytic parameters, as well the serum levels of iron, ferritin and erythropoietin were determined. All the DIPF patients studied, apart from two, had normal or subnormal haematocrit values. The patients with COPD had an inconsistant response to hypoxaemia; 12 had normal or subnormal haematocrit values and the remaining 17 were erythraemic. The mean value of erythropoietin (EPO) in both DIPF and COPD patients was significantly higher than normal. In conclusion, patients with DIPF exhibit a lack of erythropoietic response to hypoxaemia, despite the augmented erythropoietin levels. This may reflect a defective bone marrow erythropoietic response in DIPF patients. It is suggested that the pathophysiology of DIPF underlies this mechanism. PMID- 11421505 TI - Does the mode of inhalation affect the bronchodilator response in patients with severe COPD? AB - Spacing devices improve lung deposition of aerosols from metered dose inhalers (MDI) but it is sometimes difficult for dyspnoeic patients to perform maximal breaths with breath-holds needed to inhale the aerosols from them. Our aim was to determine whether the response to bronchodilators (BD) depended on the method of inhalation. We studied 20 patients with moderately severe chronic obstructive pulmonary disease (COPD) with a mean age of 68 years and a mean of forced expiratory volume in 1 sec (FEV1) of 41% predicted. In a randomized, cross-over fashion they inhaled terbutaline 1.5 mg (six puffs) followed by ipratropium 120 microg (six puffs) via MDI and nebuhaler with either two inspirations to total lung capacity and a 10-sec breath-hold per puff or with six tidal breaths per puff. Before and after BDs we measured FEV1, forced vital capacity (FVC), airways resistance using interrupter method (Rint) and 6-min walking distance (6MWD). Subsequently, we re-tested nine of these patients with the two methods of inhalation, before and after conventional doses (terbutaline 500 microg+ipratropium 40 microg), then after terbutaline 1 mg and ipratropium 80 microg and finally after nebulized terbutaline 5 mg and ipratropium 500 microg to sec whether there was a dose-dependent difference in effect between the two methods. Spirometry, slow vital capacity (SVC). inspiratory capacity and shuttle walking tests were monitored. In the original 20 patients there were highly significant improvements in all parameters after inhalers, with no significant difference between methods of inhalation. Median improvements after BDs were: FEV1 0.221 and 0.191, FVC 0.501 and 0.381 and 6MWD 40 m and 44 m, for maximal breaths and tidal breathing, respectively. For nine patients, tidal and maximal breaths produced similar effects on lung function and exercise tolerance at both doses of BDs. Nebulized BDs only improved shuttle distances slightly when compared with either method of inhalation from MDI and spacer but had no additional effect on lung function. In conclusion, in patients with moderately severe COPD, BDs given by metered dose inhaler via nebuhaler have similar effects whether given by six easy tidal breaths or the more difficult two maximal breaths with breath-hold. This holds true at small or larger doses of BD. Either method of inhaling six puffs of the BDs can be used as an effective alternative to nebulized aerosol. PMID- 11421506 TI - Protection against cold air and exercise-induced bronchoconstriction while on regular treatment with Oxis. AB - This study aimed to compare the duration of protection against exercise-induced bronchoconstriction (EIB) after inhalation of formoterol (Oxis) Turbuhaler with that of terbutaline Turbuhaler and placebo Turbuhaler in asthmatic patients treated regularly with formoterol Turbuhaler 9 microg b.i.d. and inhaled steroids. The study. performed at three centres (Goteborg and Lund, Sweden, and Trondheim, Norway), consisted of an open-label part with formoterol Turbuhaler 9 microg b.i.d. and a randomized, double-blind, cross-over part with a single dose (on top of the regular treatment) of either formoterol Turbuhaler 9 microg, terbutaline Turbuhaler 0.5 mg or placebo Turbuhaler. The patients attended the clinic six times: twice for screening visits, three times for randomized treatment and once for a follow-up visit. Patients received regular b.i.d. treatment with formoterol 9 microg for a mean period of 16 days. Formoterol gave a post-exercise fall of 12, 10, 15 and 17% in forced expiratory volume in 1 sec (FEV1) 15 min, 4, 8 and 12 h after inhalation. The differences compared with placebo (falls of 26, 22, 23 and 22%) and terbutaline (falls of 17, 18, 22 and 22%) were all statistically significant (P<0.05 for all comparisons). Patients on regular treatment with formoterol Turbuhaler 9 microg b.i.d. have a significant protection against EIB up to 12 h after inhalation of formoterol 9 microg. The protection was also significantly better than that of terbutaline Turbuhaler 0.5 mg. PMID- 11421507 TI - Nitric oxide (NO) in exhaled air after experimental ozone exposure in humans. AB - We hypothesized that ozone, a common air pollutant, potent in producing airway inflammation, would increase the production of exhaled nitric oxide (NO). If so, measurement of exhaled NO could potentially be a valuable tool in population studies of air pollution effects. Eleven healthy non-smoking volunteers were exposed to 0.2 ppm ozone (O3) and filtered air for 2h on two separate occasions. Exhaled NO and nasal NO were measured before and on five occasions following the exposures. Changes in exhaled and nasal NO after ozone exposure were adjusted for changes after air exposure. There was a slight decrease in exhaled NO (-0.6; -3.1 1.2 ppb) (median and 95% confidence interval) and of nasal NO (-57; -173-75 ppb) directly after the ozone exposure. No significant changes in exhaled or nasal NO were however found 6 or 24 h after the exposure. Within the examined group, an O3 exposure level proven to induce an airway inflammation caused no significant changes in exhaled or nasal NO levels. Hence, the current study did not yield support for exhaled NO as a useful marker of ozone-induced oxidative stress and airway inflammation after a single exposure. This contrasts with data for workers exposed to repeated high peaks of ozone. The potential for exhaled NO as a marker of oxidative stress therefore deserves to be further elucidated. PMID- 11421508 TI - Added value of co-morbidity in predicting health-related quality of life in COPD patients. AB - The extent to which a chronic obstructive pulmonary disease (COPD) patient is impaired in health-related quality of life (HRQoL) is only to a small extent reflected in clinical and demographical measures. As the influence of comorbidity on HRQoL is less clear, we investigated the added value of 23 common diseases in predicting HRQoL in COPD patients with mild to severe airways obstruction. COPD patients from general practice who appeared to have an forced expiratory volume in 1 sec/inspiratory vital capacity (FEV1/IVC) < predicted -1.64 SD, FEV1 <80% predicted, FEV1 reversibility < 12% and a smoking history, were included (n=163). HRQoL was assessed with the short-form-36 (SF-36) and the presence of comorbidity was determined by a questionnaire, which asked for 23 common diseases. All domains of the SF-36 were best predicted by the presence of three or more co morbid diseases. FEV1 % predicted, dyspnoea and the presence of one or two diseases were second-best predictors. Co-morbidity explained an additional part of the variance in HRQoL, particularly for emotional functioning (delta R2=0.11). When individual diseases were investigated, only insomnia appeared to be related to HRQoL. As HRQoL is still only partly explained, co-morbidity and other patient characteristics do not clearly distinguish between COPD patients with severe impairments in HRQoL and COPD patients with minor or no impairments in HRQoL. Therefore, it remains important to ask for problems in daily functioning and well being, rather than to rely on patient characteristics alone. PMID- 11421509 TI - Adding formoterol to budesonide in moderate asthma--health economic results from the FACET study. AB - The FACET (Formoterol and Corticosteroid Establishing Therapy) study established that there is a clear clinical benefit in adding formoterol to budesonide therapy in patients who have persistent symptoms of asthma despite treatment with low to moderate doses of an inhaled corticosteroid. We combined the clinical results from the FACET study with an expert survey on average resource use in connection with mild and severe asthma exacerbations in the U.K., Sweden and Spain. The primary objective of this study was to assess the health economics of adding the inhaled long-acting beta2-agonist formoterol to the inhaled corticosteroid budesonide in the treatment of asthma. The extra costs of adding the inhaled beta2-agonist formoterol to the corticosteroid budesonide in asthmatic patients in Sweden were offset by savings from reduced use of resources for exacerbations. For Spain the picture was mixed. Adding formoterol to low dose budesonide generated savings, whereas for moderate doses of budesonide about 75% of the extra formoterol costs could be recouped. In the U.K., other savings offset about half of the extra cost of formoterol. All cost-effectiveness ratios are within accepted cost-effectiveness ranges reported from previous studies. If productivity losses were included, there were net savings in all three countries, ranging from Euro 267-1183 per patient per year. In conclusion, adding the inhaled, long-acting beta2-agonist formoterol to low-moderate doses of the inhaled corticosteroid budesonide generated significant gains in all outcome measures with partial or complete offset of costs. Adding formoterol to budesonide can thus be considered to be cost-effective. PMID- 11421510 TI - Cyclodextrin as a potential drug carrier in salbutamol dry powder aerosols: the in-vitro deposition and toxicity studies of the complexes. AB - This research was carried out to develop a new carrier in dry powder aerosols. Two types of cyclodextrin were chosen; gamma cyclodextrin (GCD) and dimethyl-beta cyclodextrin (DMCD) as carriers in dry powder formulations. Salbutamol was used as a model drug and a control formulation containing lactose and the drug was included. A twin-stage impinger (TSI) was used to evaluate in delivery efficiency of those dry powder formulations. The toxicity of cyclodextrin complexes was investigated in the rat by monitoring blood urea nitrogen (BUN) and urinary creatinine, as well as determining haemolysis of human red blood cells. The release of salbutamol from dry powder formulations was also studied over a period of time. From the results obtained, it was found that the formulation containing GCD-enhanced drug delivery to the lower stage of the TSI (deposition = 65%) much greater than that of both formulations containing DMCD (50%) and the control formulation (40%) (P<0.05). After injecting the GCD complex BUN and creatinine levels in rats were similar to those obtained in the control while those receiving DMCD complex had higher BUN and creatinine. The haemolysis of red blood cells incubated with the DMCD complex was higher than that obtained in the GCD complex. The drug release in both formulations containing GCD and DMCD was fast (over 70% was released in 5 min) and nearly all the drug was released within 30 min. It can be concluded that GCD and DMCD are able to promote salbutamol delivery in dry powder inhaler compared to a formulation containing lactose. In addition, GCD is relatively safe in the rat if the amount of GCD in the formulation is similar to this experiment. PMID- 11421511 TI - Exhaled nitric oxide in patients with PiZZ phenotype-related alpha1-anti-trypsin deficiency. AB - There is no report of exhaled NO (eNO) in subjects with different phenotypes of alpha1-anti-trypsin (AAT) deficiency. Exhaled nitric oxide was evaluated by means of single-breath chemiluminescence analysis (fractional exhaled concentration at the plateau level [plFE(NO)]) in 40 patients with AAT deficiency. Patients were divided according to the protease inhibitor (Pi) phenotype: PiMZ/MS, n = 25; PiSZ n = 6; PiZZ, n = 9. Nineteen healthy subjects served as controls. Levels of eNO in PiZZ patients were also compared with those of subjects, without AAT deficiency (PiMM), matched for diagnosis, sex, age, smoking habit and forced expiratory volume in 1 sec (FEV1). In AAT deficiency subjects airway hyper responsiveness to methacholine (PD20 FEV1) was also assessed. plFE(NO) was significantly lower in the PiZZ group (4.5+/-1.4 ppb) than in matched PiMM subjects (8.2+/-3.8 ppb), in healthy controls (9.3+/-2.8 ppb) and in patients of other phenotypes. Dynamic lung volumes and DL(CO) were significantly lower in PiZZ than in other AAT-deficient patients. Bronchial hyper-responsiveness was not different among AAT phenotypes. These results suggest that eNO may be significantly reduced in PiZZ as compared to healthy control subjects and to AAT subjects with other phenotypes, independent of the level of airway obstruction. Whether, at least potentially, eNO may be considered as an early marker of lung involvement in AAT deficiency must be confirmed with studies on larger number of subjects. PMID- 11421512 TI - Evaluation of an inspiratory muscle trainer in healthy humans. AB - The Powerbreathe is an inspiratory muscle trainer promoted as improving inspiratory muscle strength (and consequently exercise performance) in athletes and patients with respiratory disease. No published evidence supports its efficacy. We performed a prospective randomized controlled study in which 12 normal subjects received either Powerbreathe training or sham training for a 6 week period. The primary outcome measure was diaphragm strength evaluated as twitch transdiaphragmatic pressure (Tw Pdi) but secondary outcome measures were provided by full respiratory muscle assessment and cardiopulmonary exercise testing. An advantage to training was observed when outcome was assessed by maximal static inspiratory mouth pressure (mean advantage 14.5 cm H2O, 95% CI 2.2 26.9 cm H2O, P=0.025). However. no significant difference was observed between the groups in any other parameter. In particular the deltaTw Pdi was not different between groups (mean 'advantage' 0.7 cmH2O, 95% CI- 7.0+/-5.5 cmH2O, P=0.8). The continued sale and use of the Powerbreathe device is not justified by our data. A sample size calculation showed that 234 subjects would need to be randomized to definitively refute the hypothesis that Powerbreathe improves Tw Pdi and we argue that such a study is required. PMID- 11421513 TI - Acute eosinophilic pneumonia with increased soluble ST2 in serum and bronchoalveolar lavage fluid. PMID- 11421514 TI - Equivalent therapeutic ratio, of salbutamol given by Turbuhaler and Diskus (Respir Med 2000, 94: 574-577). PMID- 11421515 TI - Prevalence of denervation in paraspinal and foot intrinsic musculature. AB - OBJECTIVE: The primary purpose of this investigation was to determine the prevalence of abnormal spontaneous activity (positive sharp waves (PSWs) and fibrillation potentials (FPs)) in selected lumbosacral paraspinal and foot intrinsic muscles in an asymptomatic healthy population. DESIGN: This was a prospective assessment of 50 individuals without history or physical findings suggestive of peripheral neuromuscular disease whereby a monopolar needle electrode was located in the unilateral L4 and L5 paraspinal as well as abductor hallucis and extensor digitorum brevis muscles. These muscles were extensively evaluated for the presence of PSWs, FPs, and fasciculation potentials. RESULTS: Ten subjects per decade from 20-59 yr and ten subjects from 60-80 yr comprised the 50 participants (28 women), resulting in a mean age of 45+/-15.9 (range, 20 76) yr. A single individual (prevalence, 2%) demonstrated fibrillation potentials in the extensor digitorum brevis, and FPs and PSWs were detected in two subjects' (4% prevalence) L4/L5 paraspinal muscles. Ninety-four percent of the subjects had fasciculation potentials in the abductor hallucis, whereas 60% had these waveforms in the extensor digitorum brevis. Only 6% of subjects had fasciculation potentials in the L4 but not L5 paraspinal muscles. All subjects demonstrated both prototypical and "atypical" appearing endplate spikes in all of the muscles examined. CONCLUSIONS: We failed to confirm the previously reported prevalence of FPs and PSWs in both the paraspinal and foot intrinsic musculature. Atypical appearing endplate spikes, however, display configurations similar to FPs and PSWs and were present in all subjects. Failure to pay close attention to the discharge rate and rhythm of endplate spikes can lead to misinterpreting these waveforms as FPs and PSWs. It is likely that the previously reported high prevalence of spontaneous activity in healthy persons resulted from not fully appreciating the similarity between innervated and denervated spontaneous single muscle fiber discharge configurations. PMID- 11421516 TI - H reflex and F wave latencies to soleus normal values and side-to-side differences. AB - OBJECTIVE: Electromyographers must reliably differentiate between H reflexes and F waves when recording from the soleus muscle in the evaluation of S1 radiculopathy. The use of F waves in root-level injuries is questioned, whereas H reflexes have shown value in the evaluation of S1 radiculopathy. We studied the relationship between the tibial H reflex and F wave latencies in the limbs of 40 subjects. DESIGN: After recording the H wave latency, we changed the gain to 200 microV/cm and increased the stimulation to supramaximal for ten additional responses without moving the recording or stimulating electrodes. We also calculated the predicted H wave latency with the standard formula. Forty subjects, mean age 32 yr, consented and participated. RESULTS: The mean of the average F wave was 1.76 ms longer than the ipsilateral H reflex latency. The mean side-to-side difference of the average F wave was 0.56 ms. The H reflex latency side-to-side difference was 0.36 ms. CONCLUSION: The findings suggest that the average F wave latencies have a predictive value in the clinical context similar to the H reflex. PMID- 11421517 TI - Comparative analysis of biofeedback and physical therapy for treatment of urinary stress incontinence in women. AB - OBJECTIVE: To compare the effectiveness of an intensive group physical therapy program with individual biofeedback training for female patients with urinary stress incontinence. DESIGN: Randomized study of two therapeutic interventions consisting of a specific physical therapy program (PT) or biofeedback training (BF) daily for 4 wk, followed by a 2-mo, unsupervised home exercise program in both groups in an outpatient clinic of a large university hospital. Forty women, referred by gynecologists for nonoperative treatment of genuine stress incontinence of mild-to-moderate severity, were included. Measurements of daytime/nocturnal urinary frequency and subjective improvement of incontinence were the main outcome measures at initial presentation, after completion of the therapy program, and at follow-up after 3 mo. Standardized examinations of digital contraction strength, speculum tests, and manometric measurements were documented as secondary outcome measures. RESULTS: In the PT group, the daytime urination frequency decreased 22% after 4 wk of therapy and 19% after 3 mo (P < 0.05) from baseline. The nocturnal urination frequency was reduced by 66% after 4 wk of therapy and 62% after 3 mo (P < 0.001). In the BF group, the daily urination frequency decreased 10% after 4 wk of therapy and 5% after 3 mo (P > 0.05). The nocturnal urination frequency declined 36% after 4 wk of therapy and 66% after 3 months (P < 0.05). Subjective assessment after 3 mo showed that in the PT group, 28% of patients were free of incontinence episodes, 68% reported improved symptoms (incontinence episodes improved by >50%), and 4% were unchanged. In the BF group, 62% were free of incontinence episodes, and 38% were improved. Results of the digital contraction strength assessments, speculum tests, and manometric measurements showed statistically significant improvement in all variables in both groups after 3 months. CONCLUSION: Four weeks of both intensive group physical therapy or individual biofeedback training followed by an unsupervised home exercise program for 2 mo are effective therapies for female urinary stress incontinence and result in a significantly reduced nocturnal urinary frequency and improved subjective outcome. Only group physical therapy resulted in reduced daytime urinary frequency. BF therapy resulted in a better subjective outcome and higher contraction pressures of the pelvic floor muscles. PMID- 11421518 TI - Progressive resistance muscle strength training of hospitalized frail elderly. AB - OBJECTIVE: To determine whether frail elderly patients recuperating from acute illnesses could safely participate in and gain appreciable improvement in muscle strength from progressive resistance muscle strength training. DESIGN: Muscle strength (one repetition maximum), functional abilities (sit-to-stand maneuver and 20-sec maximal safe gait speed), and body composition were measured before and at the conclusion of a 10-wk program of lower limb progressive resistance muscle strength training. The nonrandomized study was conducted in a 30-bed geriatric rehabilitation unit of a university-affiliated Veterans Affairs hospital and a 28-bed transitional care unit of a community nursing home. Participants included 19 recuperating elderly subjects (14 male, 5 female; 13 ambulatory, 6 nonambulatory) >64 yr (mean age, 82.8+/-7.9 yr). RESULTS: The one repetition maximum increased an average of 74%+/-49% (median, 70%; interquartile range, 38%-95%, and an average of 20+/-13 kg (P = 0.0001). Sit-to-stand maneuver times improved in 15 of 19 cases (79%). Maximum safe gait speeds improved in 10 of 19 cases (53%). Four of the six nonambulatory subjects progressed to ambulatory status. No subject experienced a complication. CONCLUSIONS: A carefully monitored program of progressive resistance muscle strength training to regain muscle strength is a safe and possibly effective method for frail elderly recuperating from acute illnesses. A randomized control study is needed to examine the degree to which progressive resistance muscle strength training offers advantages, if any, over routine posthospital care that includes traditional low-intensity physical therapy. PMID- 11421519 TI - Long-term causes of death after traumatic brain injury. AB - OBJECTIVE: To determine which causes of death are more frequent in persons with traumatic brain injury (TBI), and by how much, compared with the general population. Our focus was the period beginning 1 yr after injury. DESIGN: Subjects were 2320 Californians with long-term mental disability after a TBI at age 10 yr or more, followed up between 1988 and 1997. The units of study were person-years, each linked to the subject's age, gender, level of ambulation, time since injury, and cause of death (if any) for the specific year. Observed numbers of cause-specific deaths were compared with numbers expected according to general population mortality rates. RESULTS: Mortality was higher between 1.0 and 5.0 yr postinjury than after 5.0 yr and was strongly related to reduced mobility. Death rates were elevated for circulatory diseases, respiratory diseases, choking/suffocation, and seizures, with seizure deaths being relatively frequent, even among the most ambulatory. CONCLUSIONS: Death rates for several causes are elevated in persons with long-term sequelae of TBI. The increased risk of choking/suffocation should be of interest to caregivers. Life expectancy seems to be reduced, even for patients who are fully ambulatory. PMID- 11421520 TI - Insurance case managers' perception of quality in back pain programs: a focus study group. AB - OBJECTIVES: Insurance case managers commonly interact with physiatrists and rehabilitation programs. They influence referrals and patients' decision making. This study was designed to determine which factors affect case managers' perception of back pain program quality. DESIGN: Repeated focus group interview in a neutral facility in an urban Midwestern United States community. Subjects were two groups (n = 12 and 11) of insurance case managers employed by case management firms (large and small), insurers, and self-insured employers. Outcome measures included group and individual responses to a pre-scripted interview and were collected on tape, transcribed, and interpreted by two different persons: the independent expert interviewer and a pain psychologist. RESULTS: There was substantial agreement between the two interpreters. Both groups overwhelmingly chose physiatrists over other specialists. They emphasized timeliness, communication, functionally oriented programs, concrete program goals and time frames, physician knowledge of the legal aspects of disability, and rapid communication of patient noncompliance. CONCLUSIONS: Rehabilitation programs may strive to meet many of these qualities but, in doing so, should be aware that the legal and ethical roles of case managers differ from that of clinicians. PMID- 11421521 TI - The influence of insurance case managers. PMID- 11421522 TI - Assistive technology outcomes research: challenges met and yet unmet. AB - This article highlights the special requirements, achievements, and yet unmet challenges of assessing the outcomes of assistive technology services. The current status of this research is considered from the standpoint of developmental stages that seem to characterize many areas of outcomes research. Those stages include exhortation, sober appraisal, infrastructure building, and "getting on with it." The status of measuring assistive technology outcomes is described, and efforts to develop new measures are critically reviewed. Three as yet unmet challenges are discussed that are faced alike by assistive technology outcomes research and by rehabilitation outcomes research in general. They are as follows: (1) operationalizing a multiple-stakeholder approach to outcomes research; (2) formulating adequate treatment theories; and (3) creating shared databases. PMID- 11421523 TI - Nonoperative treatment of an interosseous ganglion cyst. AB - Ganglion cysts of the knee are being reported more frequently secondary to an increased rate of magnetic resonance imaging studies. Although knee pain is the impetus for imaging, ganglion cysts are often incidental findings. Nonoperative treatment is a successful therapeutic option. We report a patient with variable pain presentations over the course of her treatment. The pattern of complaints pointed to different primary etiologies about the knee, but all were common to an interosseous ganglion cyst. A stepwise assessment and expansion of the differential diagnosis allowed for appropriate utilization of modalities and limited morbidity with nonoperative therapy. PMID- 11421524 TI - Improvement of sleep apnea in a patient with cerebral palsy. AB - Intrathecal baclofen (ITB) can reduce spasticity in adults and children with cerebral palsy. Benefits of ITB therapy include improved Ashworth scores, activities of daily living, and mobility. The impact of ITB therapy on sleep apnea in patients with cerebral palsy has not been reported. This case report describes a 29-yr-old female with mixed spastic athetoid quadriparetic cerebral palsy with dystonia, gross motor function IV, who had sleep apnea, requiring nightly continuous positive airway pressure. She received ITB with the goal to improve her wheelchair positioning and decrease her excessive movements. After the initiation of the ITB, reduction of her spasticity and dystonia was noted, as well as improvement of her sleep apnea. This case suggests that ITB therapy may improve respiratory function through reduction of respiratory muscle spasticity. PMID- 11421525 TI - Isolated sural neuropathy presenting as lateral ankle pain. AB - A case of job-related, unilateral traumatic sural neuropathy causing severe lateral ankle pain and impaired work performance for a 26-yr-old female grocery clerk is reported. This diagnosis is made both clinically and electrophysiologically. We review the pertinent electrophysiologic features, anatomy, and clinical findings in our patient with an isolated sural neuropathy. A review of the literature demonstrates that trauma is the most common cause of this unusual isolated neuropathy. Despite its rare occurrence, it should be considered in patients who present with lateral ankle pain and concomitant loss of sensation in the sural nerve distribution. The establishment of a neuropathic origin assists with management strategies that will differ from the more common musculoskeletal causes of lateral ankle pain. After an appropriate diagnosis and treatment, an excellent outcome resulted for our patient. PMID- 11421526 TI - Conducting clinical trials to establish drug efficacy in chronic pain. AB - Design of clinical trials to establish drug efficacy in chronic pain is a complicated issue, and numerous factors must be considered in identifying an optimal study design. Investigators should begin by identifying a focused and testable research question, with the outcome variables operationalized in a way that allows appropriate quantitative analysis. The prospective, randomized, placebo-controlled, doubled-blind study using validated quantitative measures is considered the optimal clinical trial design. Within this general study type, the between-subjects design has less statistical power than does the crossover design, in which the patient serves as his or her own control. However, potential problems with the drug effects from the first condition carrying over into, and confounding, the second drug condition present a noteworthy limitation that must be addressed through adequate washout periods and statistical control if crossover designs are used. Retrospective designs may be useful primarily to take advantage of samples of convenience for development of pilot data that provide the basis for conducting better-controlled prospective studies. Sample selection issues must be considered during study design, including the sample size required (based on statistical power analysis), appropriate inclusion and exclusion criteria, and likely availability of qualifying patients. There are numerous statistical options for analyzing data that must be selected based on whether data are parametric or nonparametric, whether within-subject (crossover) or between-subject comparisons are used, and whether baseline values of outcome measures affect the degree of change in these measures over the course of the study. Involvement of a biostatistical consultant is recommended during all phases of clinical trials. PMID- 11421527 TI - Prevalence of osteoporosis in women: variation with skeletal site of measurement of bone mineral density. AB - OBJECTIVE: To determine retrospectively the prevalence of osteoporosis in a referral population of female patients and to compare the sensitivity for diagnosing osteoporosis by dual-energy x-ray absorptiometry (DXA) measurements of bone mineral density (BMD) at multiple skeletal sites. METHODS: We studied the data from 625 consecutive women (mean age, 57.3 +/- 13.9 years), who had been referred to our center for lumbar spine (anteroposterior [AP] and lateral region) and hip (femoral neck [FN], Ward's triangle [WT], trochanter, intertrochanteric region, and total hip) BMD measurements with use of DXA (Hologic QDR-2000) between June 1994 and July 1998. RESULTS: Osteoporosis (based on the World Health Organization definition--T-score of -2.5 or lower for BMD) was diagnosed by DXA at the following sites: AP spine in 21.7%, lateral spine in 43.2%, FN in 33.6%, WT in 49.1%, trochanter in 26.1%, intertrochanteric region in 25.9%, and total hip in 28.4% of study patients. Significant site differences were found in the prevalence of osteoporosis between the lateral and AP spine (P < 0.001), as well as between WT and the FN, trochanter, intertrochanteric region, and total hip (P < 0.001). In a subgroup of 71 women, forearm (ultradistal radius and radius 1/3 region) BMD results indicated low sensitivity for diagnosing osteoporosis, similar to that seen at the AP spine, trochanter, and intertrochanteric region. Not surprisingly, the prevalence of osteoporosis increased with advancing age (15.5% in patients younger than 50 years, in comparison with 59.6% in those older than 69 years of age). The frequency of misclassification of patients (osteoporosis at one site and normal BMD at another) with use of the seven measurement sites was 16.6% (104 of the 625 patients). CONCLUSIONS: For diagnosis of osteoporosis, DXA BMD measurements are significantly more sensitive at the lateral spine than at the AP spine, as well as at WT than at the FN, trochanter, intertrochanteric region, and total hip sites. PMID- 11421528 TI - Testosterone treatment in hypogonadal men: prostate-specific antigen level and risk of prostate cancer. AB - OBJECTIVE: To assess prostate-specific antigen (PSA) levels in hypogonadal men after testosterone replacement by three different methods and attempt to determine any possible relationship between hypogonadism and prostate cancer in this study population. METHODS: A total of 90 consecutive men who had erectile dysfunction and were found to have hypogonadism were monitored with digital rectal examination (DRE) and measurement of PSA levels before and after testosterone replacement therapy. The patients were treated with one of three options: (1) testosterone enanthate by intramuscular injections, 200 or 300 mg every 2 or 3 weeks (N = 25); (2) testosterone nonscrotal patches, 5 mg daily (N = 16); or (3) clomiphene citrate, 50 mg orally three times a week, in patients with functional secondary hypogonadism (N = 49). Treatment was continued for 2 to 3 months, after which PSA levels were reassessed. Patients with suspicious results on DRE and increased PSA levels before or after treatment with testosterone underwent prostate biopsy. For statistical analysis, patients were categorized into two age-groups--40 to 60 years old and 61 to 80 years old. RESULTS: With all methods of testosterone replacement, PSA levels increased in both age-groups. Endogenous testosterone elevation from clomiphene stimulation raised PSA levels the highest, and testosterone patches yielded the least PSA response. Ten men underwent biopsy of the prostate. In one patient, a nodule was found on DRE; the other nine men underwent biopsy because of suspicious PSA levels. Of these patients, two were found to have adenocarcinoma, and a third man who underwent rebiopsy was also found to have cancer. Therefore, 3 of the 90 patients (3.3%) had prostate cancer. CONCLUSIONS: PSA levels increased in response to all types of testosterone replacement, regardless of whether the testosterone level was raised endogenously or exogenously. PSA levels are inappropriately low in hypogonadal men and may mask an underlying cancer. Determining PSA levels before and after testosterone treatment is recommended. Elevated PSA levels before or after testosterone therapy should prompt performance of a urologic evaluation for possible prostate biopsy. PMID- 11421529 TI - Role of thyroid-stimulating immunoglobulin in aggressiveness of well differentiated thyroid cancer. AB - OBJECTIVE: To assess whether a relationship exists between thyroid-stimulating antibodies and increased aggressiveness of thyroid cancer. METHODS: We analyzed clinical, histologic, and biochemical data, including thyroid-stimulating antibodies, from 26 patients (24 women and 2 men) who had had well-differentiated thyroid carcinoma for 1 to 5 years and had undergone total thyroidectomy and radioactive iodine ablative therapy. For analysis, the overall study cohort was divided into two groups: group 1 (N = 16), with stable disease and no evidence of metastatic activity, and group 2 (N = 10), with aggressive disease and substantiated metastatic involvement. RESULTS: The thyroid-stimulating antibodies ranged from 92 to 129% in group 1 and from 95 to 118% in group 2. Thus, both study groups had thyroid-stimulating antibody levels within the normal range (normal, <130%). CONCLUSIONS: Apparently, thyroid-stimulating antibodies had no contributory role in the growth of the metastatic lesions in the 10 patients with aggressive disease. Further studies should be undertaken to investigate other potential factors involved in stimulating the progression of thyroid cancer. PMID- 11421530 TI - A cluster of cases of factitious Cushing's syndrome. AB - OBJECTIVE: To review the patient profiles, laboratory data, and diagnostic approaches in factitious administration of glucocorticoids. METHODS: Four cases of surreptitious use of glucocorticoids are presented. Clinical and laboratory data as well as imaging studies are summarized. Pertinent case reports in the literature are reviewed. RESULTS: We report four cases of surreptitious use of glucocorticoids encountered within a 2-year period. All four patients were women without significant psychiatric histories. In three patients, the question of factitious Cushing's syndrome was suspected because of physical evidence or symptoms of Cushing's syndrome (or both) in the setting of suppressed cortisol levels. The fourth patient had undetectable cortisol levels in both serum and 24 hour urine samples but did not have signs or symptoms of adrenal insufficiency. In three cases, the diagnosis was confirmed by direct measurement of synthetic glucocorticoids in the patient's urine or serum. The fourth case was diagnosed by correlating increased cortisol levels with decreased precursor adrenal steroids. CONCLUSIONS: Exogenous corticosteroid use in the absence of a medical indication poses a serious risk to a patient. This possibility should be considered in patients with signs and symptoms consistent with Cushing's syndrome but with low serum and urinary cortisol levels. Similarly, this diagnosis should be suggested in patients without symptoms of adrenal insufficiency and with low cortisol levels. Laboratory measurement of synthetic steroids can be helpful in confirming the diagnosis. PMID- 11421531 TI - Successful pregnancy in a woman with acromegaly treated with octreotide. AB - OBJECTIVE: To report a successful pregnancy in a 43-year-old woman with acromegaly and treatment during pregnancy with the somatostatin analogue octreotide, to review the physiologic aspects of acromegaly during pregnancy, and to discuss treatment options. METHODS: A case report is presented, and the current literature is reviewed. RESULTS: In a 40-year-old woman with acromegaly and secondary amenorrhea, menstrual periods resumed after treatment with octreotide. At age 43 years, she became pregnant and decided to continue octreotide therapy because of the relatively large size of the tumor and severe headaches when use of the medication was discontinued. Growth hormone and insulin like growth factor I levels remained normal throughout pregnancy, and a normal full-term infant was born. Several months after delivery, severe cholecystitis developed, and the patient underwent laparoscopic cholecystectomy. CONCLUSIONS: This case report describes the oldest documented woman with acromegaly who has completed a pregnancy and only the second patient treated with octreotide for the entire pregnancy. During a normal pregnancy, placental growth hormone suppresses pituitary growth hormone secretion; in patients with acromegaly, pituitary growth hormone secretion remains autonomous. Treatment of acromegaly with octreotide, bromocriptine, irradiation, or surgical intervention may restore menses and fertility to women of reproductive age. Bromocriptine has been used throughout pregnancy without teratogenic effects in many patients. Octreotide has been used during the first trimester of pregnancy in seven other women with acromegaly (throughout pregnancy in one of those patients) and for the entire pregnancy in one woman with a thyrotropin-secreting pituitary tumor; no adverse effects were noted. Lanreotide was also used in one patient briefly during the first trimester without adverse consequences. Although the outcomes of the current and previous pregnancies have been favorable, insufficient data are available to recommend the routine use of octreotide in all pregnant women with acromegaly. PMID- 11421532 TI - Human recombinant DNA insulin-induced lipoatrophy in a patient with type 2 diabetes mellitus. AB - OBJECTIVE: To present the first case of lipoatrophy occurring in a patient with type 2 diabetes mellitus who was treated with human recombinant DNA (rDNA) derived insulin as her only exposure to exogenous insulin. METHODS: We describe the clinical findings in a 47-year-old woman with lipoatrophy after injection of human rDNA insulin for type 2 diabetes, and we review the related literature. RESULTS: Although a few case reports have documented lipoatrophy in patients with type 1 diabetes treated with human insulin, to our knowledge no previous reports have described patients with type 2 diabetes in whom lipoatrophy developed after injection of human rDNA insulin. This was our patient's first exposure to exogenous insulin. No definite factor was found to be contributory to the development of lipoatrophy in this case, even though several factors may be related. CONCLUSIONS: This is the first published case report of lipoatrophy occurring in a patient with type 2 diabetes mellitus who received human rDNA insulin as her only exposure to exogenous insulin. PMID- 11421533 TI - A case of unilateral adrenal hyperplasia: the diagnostic dilemma of hyperaldosteronism. AB - OBJECTIVE: To report the successful laparoscopic surgical management of a case of right unilateral adrenal hyperplasia. METHODS: We present a case of unilateral adrenal hyperplasia and review the pertinent literature. The effectiveness of surgical treatment of unilateral adrenal hyperplasia is evaluated. RESULTS: A 50 year-old man had a 7-year history of hypertension, which had been treated with antihypertensive medications. On initial assessment, he was taking long-acting diltiazem, 240 mg twice a day, and his blood pressure was 150/84 mm Hg. He was noted to have recurrent hypokalemia, low plasma renin activity, increased plasma aldosterone concentration, and high urinary aldosterone levels. Magnetic resonance imaging of the adrenal glands revealed a normal left adrenal gland and fullness of the right adrenal gland. Selective adrenal vein catheterization showed findings consistent with a right-sided adrenal gradient. Surgical removal of the right adrenal gland laparoscopically alleviated his symptoms and cured his hypertension and hypokalemia. Pathologic examination of the right adrenal gland revealed diffuse hyperplasia of the zona fasciculata and zona reticularis. Plasma aldosterone, renin, and potassium levels returned to normal postoperatively, and his blood pressure declined to 120/70 mm Hg. At 1-year follow-up, the patient remained normotensive without the need for antihypertensive agents. CONCLUSIONS: This rare type of hyperaldosteronism, known as unilateral adrenal hyperplasia, is difficult to categorize not only because it fails to typify a conventional subtype but also because it is difficult to confirm the pathologic diagnosis. Recognition of the variety of provocative, biochemical, and imaging results that may be seen in this disorder is important in making the correct diagnosis. Determining unilaterality of disease by selective adrenal vein sampling can result in a cure of hypertension and hypokalemia by surgical resection. PMID- 11421534 TI - Diabetic ketoacidosis in association with acanthosis nigricans. AB - OBJECTIVE: To report the occurrence of diabetic ketoacidosis in three patients with acanthosis nigricans. METHODS: Case reports with clinical and laboratory data are presented, and the syndrome of acanthosis nigricans is discussed. RESULTS: Three obese male patients--two American Indians and one Polynesian man- sought medical attention because of symptoms of diabetic ketoacidosis and were noted to have acanthosis nigricans. No factor was identified that could have precipitated the diabetic ketoacidosis, which was resolved by treatment with insulin. CONCLUSIONS: The unexpected association of diabetic ketoacidosis and acanthosis nigricans is documented in these three cases. The hyperinsulinemia associated with acanthosis nigricans results from insulin resistance, and when insulin resistance occurs in combination with impaired insulin secretion, diabetic ketoacidosis can result. PMID- 11421535 TI - Using bone mineral density T-scores to diagnose postmenopausal osteoporosis. PMID- 11421536 TI - Testosterone treatment in older men: effects on the prostate. PMID- 11421537 TI - Thyroid cancer and Graves' disease: the controversy ten years later. PMID- 11421538 TI - Efficacy of insulin pump therapy: mealtime delivery is the key factor. AB - OBJECTIVE: To investigate, in a clinical setting, the effect of implementation of continuous subcutaneous insulin infusion (CSII) on control of plasma glucose and to identify factors associated with improved glycemic control in patients with type 1 diabetes mellitus. METHODS: Nineteen patients (16 women and 3 men) with type 1 diabetes were studied retrospectively. Their mean age was 42.6 years (range, 30 to 58), and the mean duration of diabetes was 21 years. The subjects underwent follow-up for a mean of 14 months after conversion to CSII therapy. With use of paired t tests, pre-CSII and follow-up data were evaluated relative to changes in weight, insulin dosing, and glycosylated hemoglobin (HbA(1c)). RESULTS: At follow-up, the total daily dose of insulin had decreased by 18%, from a baseline mean value of 45.2 IU to 37.1 IU (P = 0.02). HbA(1c) was reduced from 8.4% to 7.7% (P<0.01). The total daily insulin-to-weight ratio also significantly decreased from 0.66 IU/kg to 0.53 IU/kg (P<0.05). Before insulin pump use, the regular/NPH insulin ratio was 0.5 IU; at follow-up, the pump bolus/basal insulin ratio was 1.0 IU (P = 0.02). No weight gain was observed; the mean weight of the study patients decreased 0.2 kg, from 69.4 kg at baseline to 69.2 kg at follow-up (not significantly different). CONCLUSION: In a clinical setting, CSII therapy in patients with type 1 diabetes improves glycemic control and lowers the total daily basal insulin dose without affecting weight. Improved glycemic control was associated with a shift in insulin therapy from a high percentage of intermediate acting insulin to a greater percentage of insulin administered in a meal associated bolus form. This study emphasizes the importance of mealtime insulin adjustment for tight glycemic control in patients using CSII therapy. Future studies evaluating the benefits of decreased total insulin and an increased bolus/basal insulin ratio may be important in helping to understand how to avoid long-term complications of diabetes. PMID- 11421539 TI - Thyroid peroxidase antibodies in Mexican-born healthy pregnant women, in women with type 2 or gestational diabetes mellitus, and in their offspring. AB - OBJECTIVE: To search for differences in the frequency of thyroid peroxidase antibodies (TPO-Ab) among 150 pregnant Mexican women who were healthy, had type 2 diabetes mellitus (DM), or had gestational diabetes mellitus (GDM). METHODS: Fifty healthy women, 50 women with type 2 DM, and 50 women with GDM were studied at delivery. In addition, 142 of their offspring were included in the study. TPO Ab were determined by enzyme immunoassay, and total triiodothyronine, free thyroxine, and thyroid-stimulating hormone (thyrotropin) were measured by radioimmunoanalysis. RESULTS: TPO-Ab were < or = 70 U/mL (negative) in 50% of the healthy women and in 60% and 60% of women with type 2 DM and GDM, respectively (no significant difference). TPO-Ab were 71 to 250 U/mL (slightly positive) in 40% of healthy women and in 30% and 34% of women with type 2 DM and GDM, respectively (no significant difference). TPO-Ab were > or =251 U/mL (strongly positive) in 10% of healthy women and in 10% and 6% of women with type 2 DM and GDM, respectively. One healthy woman had subclinical hypothyroidism, and the rest were euthyroid. The newborn offspring of these Mexican women were euthyroid and had similar frequencies of TPO-Ab (all had TPO-Ab <250 U/mL). CONCLUSION: (1) The frequency of TPO-Ab > or =251 U/mL was similar in pregnant Mexican women with GDM in comparison with those who were healthy or had type 2 DM. (2) The similar high frequencies of slightly positive TPO-Ab in the three groups of pregnant women can partially be explained by the existence of an ethnic factor, the very strong family history of DM in a substantial percentage of them, and the use of a more sensitive and specific assay for detection of TPO-Ab. PMID- 11421540 TI - Pheochromocytoma: clinical observations from a Brooklyn tertiary hospital. AB - OBJECTIVE: To report eight cases of pheochromocytoma, diagnosed and treated at our tertiary hospital during a 42-month period. METHODS: We review clinical manifestations as well as historical and family data. Biochemical and radiologic methods for diagnosis and tumor localization at our institution were compared with methods at other teaching centers. RESULTS: Among 4,180 patients with essential hypertension encountered during a 42-month interval, 8 had pheochromocytoma. The most common clinical findings were diaphoresis, chest or abdominal pain, palpitations, headaches, and nausea. Clinical features were remarkable only for the significant correlation between urinary excretion of vanillylmandelic acid and tumor volume (r = 0.925; P<0.01). Only one patient, with a history of medullary thyroid carcinoma, had multiple endocrine neoplasia. No unusual familial concentration of pheochromocytomas was found in our community, and our diagnostic techniques were similar to those reported from other institutions. We calculated an average annual incidence rate of 0.5 per 100,000 person-years, a figure comparable to that reported at the Mayo Clinic in Rochester, Minnesota. CONCLUSION: Because our institution does not see the volume of referral patients as does the Mayo Clinic, we suggest that the frequency of pheochromocytoma may be underestimated in the patient population that we serve. PMID- 11421541 TI - Benefits of computerized outcome analysis in diabetes management. AB - OBJECTIVE: To observe the effects of a computerized relational database designed to measure outcomes and enhance adherence to recommended diabetes management guidelines in a specialty private practice setting. METHODS: DiaTrends, a software program specifically designed to monitor the processes and outcomes of diabetes care, was introduced in a private endocrinology practice in 1996. Information on all patients with diabetes (N = 1,096) was recorded in the database. Reports ("queries") were developed to monitor the adherence to recommended guidelines and to assess the outcomes of care for patients with diabetes treated with glucose-lowering medications. Glycosylated hemoglobin (HbA(1c)) and lipid control were selected from the many outcomes and processes of care monitored as the basis of this report. RESULTS: Data were collected on 1,096 patients between 1996 and 1998. We were able to monitor adherence to published diabetes guidelines and to document the outcomes. Parallel trends toward improvement were noted in both HbA(1c) and lipid variables. The average HbA(1c) achieved for the entire patient population was 7.3%. CONCLUSION: Using a computerized database to monitor glycemic control and lipid management is effective in evaluating the outcomes of care and provides a focused approach that enhances diabetes care. These improvements in metabolic outcomes likely reflect a change in the practice patterns of the physicians as a consequence of using this computerized database system. PMID- 11421542 TI - Pigmented adrenal hyperplasia with myelolipomatous changes and bilateral testicular enlargement in an untreated man with 21-hydroxylase deficiency. AB - OBJECTIVE: To report the occurrence of a giant left adrenal tumor and bilateral testicular masses (adenomatous hyperplasia of Leydig cells) in a young man with congenital adrenal hyperplasia due to 21-hydroxylase deficiency. METHODS: The clinical, radiologic, endocrinologic, and pathologic features of this case are correlated with the findings in the literature. RESULTS: The interesting elements in this case are the rare pathologic features of the left adrenal lesion (pigmented adrenal hyperplasia with myelolipomatous changes) and the association with the infrequent testicular adrenal rest tumors. The absence of enlargement of the right adrenal gland was unexplained. CONCLUSION: The presence of these two rare complications seemed to be associated with poor adherence to medical treatment recommendations for congenital adrenal hyperplasia. PMID- 11421543 TI - Thyroid carcinoma and Graves' disease. AB - OBJECTIVE: To describe an unusual patient with thyroid carcinoma who had metastatic pulmonary nodules that regressed transiently during the active phase of Graves' disease. METHODS: The clinical, pathologic, and laboratory findings in a female patient with thyroid carcinoma, pulmonary lesions, and Graves' ophthalmopathy are presented. In addition, the patient's clinical course and results of serial studies are reviewed. RESULTS: A 62-year-old woman underwent excision of a follicular carcinoma of the thyroid and then total thyroidectomy in 1984. Two years later, chest radiography disclosed multiple pulmonary lesions; open-lung biopsy revealed follicular carcinoma. Thyroglobulin determinations continued to increase, and computed tomography of the chest showed an increase in the size and number of pulmonary nodules. Clinically, the patient remained unchanged until 4 years postoperatively, when she noted rapid onset of diplopia, proptosis, and diminished visual acuity. During this time, a thyroid-stimulating immunoglobulin assay was strongly positive, thyroglobulin levels were decreased, and computed tomography and radiography of the chest showed evidence of tumor regression. Orbital decompression partially restored vision, and the inflammatory signs decreased gradually. Concurrently, the thyroid-stimulating immunoglobulin level declined, the thyroglobulin level increased, and the pulmonary lesions increased in size and number. The patient ultimately died of metastatic disease. CONCLUSION: The reduction in size of the pulmonary nodules and decrease in thyroglobulin levels when the thyroid-stimulating immunoglobulin level was high and the regrowth of lesions when it declined suggest a causal relationship. The hypothesis of an operative autoimmune factor cannot be substantiated without serologic evidence in this case but should prompt physicians to search for other such occurrences. PMID- 11421544 TI - Isolated familial hyperparathyroidism with a novel mutation of the MEN1 gene. AB - OBJECTIVE: To describe two cases of familial hyperparathyroidism with genetic analysis and report a new gene mutation. METHODS: Clinical and laboratory details of a brother and sister with hyperparathyroidism and nephrolithiasis are presented. The family history is reviewed, and results of direct sequencing analysis for mutations in the MEN1 gene are discussed. RESULTS: Isolated familial hyperparathyroidism is uncommon. Whether this condition is a partial expression of multiple endocrine neoplasia type I, a well-known genetic disorder, or an independent entity is being debated and studied. Two cases of hyperparathyroidism were diagnosed in a brother and a sister from a family that has expressed isolated hyperparathyroidism in several family members as the only manifestation thus far. A novel genetic mutation involving a CGG to CGT alteration in codon 219 of the MEN1 gene was discovered; to our knowledge, this particular variation has not been reported previously. CONCLUSION: In patients with isolated familial hyperparathyroidism, a search for a mutation in the MEN1 gene may eventually help clarify this uncommon familial condition. PMID- 11421545 TI - Inflammation, insulin resistance, infection, diabetes, and atherosclerosis. AB - OBJECTIVE: To review the relationship between poor control of blood glucose in patients with diabetes and potential for occurrence of the acute coronary syndrome. METHODS: The role of advanced glycosylated end products in endothelial dysfunction is discussed, and the possible contributions of inflammation and infection in the rupture of atheromatous plaques are described. RESULTS: Hyperglycemia predisposes to infection by decreasing the efficacy of leukocytes and allowing increased formation of advanced glycosylated end products. The associated endothelial permeability facilitates bacterial infiltration of atheromatous plaques. Reported associations between chronic infection and coronary artery disease have involved primarily Chlamydia pneumoniae. The insulin resistance syndrome is also characterized by compromised endothelial function, which may predispose patients to unstable plaques and cardiac events. CONCLUSION: In patients with diabetes, poor glycemic control may lead to excessive advanced glycosylated end products, inflammation and infection of atheromatous plaques, and plaque rupture that can cause angina or myocardial infarction. PMID- 11421546 TI - Insulin pump therapy: no longer a fad. PMID- 11421547 TI - Thyroid peroxidase antibodies and pregnancy. PMID- 11421548 TI - Clarification of the terms accredit, certify, credential, and verify. PMID- 11421549 TI - Follicular variant of papillary thyroid carcinoma: a comparative study of histopathologic features and cytology results in 141 patients. AB - OBJECTIVE: To characterize the histopathologic features of follicular variant of papillary thyroid carcinoma (FVPC) and its cytology results on fine-needle aspiration (FNA) biopsy and compare them with those of papillary thyroid carcinoma (PC). METHODS: We searched the University of Massachusetts Medical Center pathology database for all surgical specimens associated with a diagnosis of FVPC or PC between January 1992 and February 1998 and reviewed the related pathology reports. In addition, the associated preoperative FNA results were analyzed. RESULTS: On initial assessment, FVPC was associated with a significantly lower incidence of cervical lymph node metastatic involvement in comparison with PC (5.6% versus 35.7%; P<0.001). Even though the mean size of FVPC was larger than that of PC (2.57 cm versus 1.75 cm; P<0.05), FVPC showed a lower incidence of thyroid capsule invasion (5.6% versus 11.4%), infiltrative resection margins (2.8% versus 20.0%; P = 0.01), local soft tissue invasion (7.0% versus 25.7%; P<0.005), and multicentricity (25.4% versus 47.1%; P<0.01). Lymphocytic thyroiditis was a common feature of both FVPC (36.6%) and PC (35.7%). FNA biopsy revealed the presence of malignant cells in 9.8% of patients with FVPC in comparison with 67.5% of patients with PC. Most cytology specimens of FVPC (58.8%) were interpreted as suspicious for a malignant lesion or as a follicular neoplasm. CONCLUSION: FVPC is associated with a significantly lower incidence of cervical lymph node metastatic lesions and invasive histologic features than is PC. Long-term prospective clinical studies are needed to determine whether these findings translate into a more benign natural history for this variant of PC. Results of FNA biopsy in FVPC are more commonly interpreted as suspicious rather than malignant; this factor has major implications for preoperative planning. PMID- 11421550 TI - Effect of immediate hemoglobin A1c results on treatment decisions in office practice. AB - OBJECTIVE: To assess the effect of an immediately available hemoglobin A1c (HbA1c) result on glycemic control and physician decisions about pharmacologic therapy in an office practice. METHODS: In a 1-year retrospective review of medical records, HbA1c results were analyzed in 115 patients beyond the age of 65 years, who had type 2 diabetes and were referred for the first time to a private endocrinology practice between April 1, 1997, and March 31, 1998. These patients were classified into two groups: group A (N = 93, insured by standard Medicare) had immediate HbA1c results (during the patient encounter) and group B (N = 22, insured by Medicare health maintenance organization [HMO]) had commercial laboratory HbA1c results available within 2 to 3 days. We reviewed the changes in the HbA1c level during the 12-month period and the presence or absence of a change in therapy at each visit. HbA1c levels were measured by ion-exchange low pressure liquid chromatography in group A and by one of three capitated commercial laboratories (depending on HMO contracts) in group B. RESULTS: At the end of the 12 months, the mean HbA1c decrease was 1.03 +/- 0.33% in group A and 0.33 +/- 0.83% in group B. During the first visit, 52% of the patients in group A had pharmacologic treatment interventions, whereas only 27% in group B had such interventions. CONCLUSION: Rapid availability of the HbA1c results during the clinical encounter improves the ability of the physician to make appropriate therapeutic decisions and results in improved glycemic control. PMID- 11421551 TI - Role of ultrafast Papanicolaou-stained scrape preparations as an adjunct to frozen sections in the surgical management of thyroid lesions. AB - OBJECTIVE: To assess the value of intraoperative cytology on scrape preparations of thyroid lesions. METHODS: We conducted a retrospective review of 73 cases of various thyroid lesions in which ultrafast Papanicolaou-stained scrape preparations were performed. Frozen sections were performed in 19 cases (26%). Fine-needle aspiration diagnosis was available in 65 cases. The correlation between the scrape diagnosis and the final diagnosis was analyzed. RESULTS: The agreement rate between the scrape diagnosis and the final diagnosis was 89%. All cases diagnosed as suspicious for papillary thyroid carcinoma on fine-needle aspiration were definite for malignant papillary carcinoma on scrape preparations. Three cases from this category were diagnosed as papillary carcinoma on both frozen section and scrape preparations, leading to total thyroidectomy. CONCLUSION: We conclude that intraoperative consultation is useful for thyroid lesions diagnosed as suspicious for papillary thyroid carcinoma on fine-needle aspiration and that ultrafast Papanicolaou-stained scrape preparations can increase the efficacy of thyroid frozen sections and obviate a second procedure for completion thyroidectomy. PMID- 11421552 TI - Effects of iodized salt consumption on goiter prevalence in Isfahan: the possible role of goitrogens. AB - OBJECTIVE: To evaluate the success of the Iranian Iodine Deficiency Disorders Committee in achieving World Health Organization (WHO) goals for reducing the prevalence of goiter in children by adding iodine to table salt beginning in 1989. METHODS: In 1997, 8,000 male and female 6- to 18-year-old students were selected by cluster sampling in schools of Isfahan. Their thyroids were examined by four endocrinologists, and goiter was staged on the basis of the WHO classification. As an index of iodine consumption, urinary iodine concentrations were measured in 3,000 students. RESULTS: Goiter was observed in 62% of the students. Of the overall study group, 94% had sufficient iodine consumption (urinary iodine concentration of 10.0 microg/dL or more). Of those students who had sufficient iodine intake, 63.2% had goiter. Of the 6% of students with iodine deficiency, 5% had mild, 0.9% had moderate, and only 0.1% had severe iodine deficiency. Goiter was absent in half of the students with severe iodine deficiency. The prevalence of goiter in 6- to 10-year-old children was 65%. CONCLUSION: Despite sufficient iodine intake, the prevalence of goiter is still high in Isfahan City. Apparently, either this high prevalence has no relationship to iodine deficiency and possibly other unknown goitrogens are involved in the pathogenesis of goiter in Isfahan or the period of iodine intake has been too brief to affect thyroid sizes. Inasmuch as goiter prevalence is also high in the 6- to 10-year-old children, who have had iodized salt available for most of their lives, the second option is less probable. Another possibility is an increased rate of autoimmune thyroid diseases (because of iodine repletion) that resemble goiter during their early stages. PMID- 11421553 TI - Use of recombinant human thyrotropin in a complicated case of metastatic papillary thyroid carcinoma. AB - OBJECTIVE: To describe a complicated case of metastatic papillary thyroid carcinoma and the use of recombinant human thyrotropin (thyroid-stimulating hormone) (rhTSH) in conjunction with treatment with radioactive iodine (131I). METHODS: We present a detailed case report and discuss the use of rhTSH in this setting. RESULTS: A 62-year-old Filipino man with a history of incompletely treated papillary thyroid cancer sought medical assistance in the emergency department because of weakness of the right upper extremity. Imaging studies showed a large mediastinal mass in association with C7 vertebral body erosion and spinal cord impingement. A cervical lymph node biopsy revealed papillary thyroid carcinoma, follicular variant. Further imaging studies disclosed widely metastatic disease. The patient subsequently had a complicated treatment course and received rhTSH in conjunction with 131I treatment. CONCLUSION: This unusual case demonstrates possible complications of metastatic papillary thyroid carcinoma along with innovations in treatment options, including the use of rhTSH to stimulate the uptake of radioiodine by thyroid cancer cells. PMID- 11421554 TI - Isolated adrenocorticotropic hormone deficiency, thyroid autoimmunity, and transient hyperprolactinemia. AB - OBJECTIVE: To describe a case of isolated adrenocorticotropic hormone (ACTH) deficiency associated with thyroid autoimmunity, subclinical hypothyroidism, and transient hyperprolactinemia. METHODS: We present a detailed case report, including results of laboratory studies and magnetic resonance imaging, and discuss potential contributing factors in this setting. RESULTS: In a 23-year-old woman with isolated ACTH deficiency accompanied by thyroid autoimmunity (Hashimoto's thyroiditis), subclinical primary hypothyroidism, and hyperprolactinemia, magnetic resonance imaging of the pituitary showed normal findings but dynamic stimulation testing of the pituitary gland indicated an isolated ACTH deficiency with intact growth hormone and gonadotropin secretory reserves. The cortisol response to the short ACTH stimulation test was subnormal. Therapy with prednisolone (5 mg/day) and levothyroxine (100 microg/day) was initiated. Results of thyroid function tests were normalized after 1 month, the prolactin level decreased to normal after 1 year, and titers of thyroid autoantibodies decreased substantially after 1.5 years of treatment. CONCLUSION: The correction of the related glucocorticoid deficiency resulted in resolution of the hyperprolactinemia and a decrease in titers of thyroid autoantibodies. PMID- 11421555 TI - Papillary carcinoma of the thyroid with manifestations resembling Graves' disease. AB - OBJECTIVE: To describe a case of papillary carcinoma of the thyroid that mimicked a diffuse toxic goiter. METHODS: We present a detailed case report, review related cases in the literature, and discuss the possible role of autoantibodies in promoting the growth of papillary thyroid tumor. RESULTS: A 28-year-old woman had a 6-month history of weight loss, palpitations, increased anxiety, and enlargement of her thyroid gland. Laboratory studies confirmed the presence of hyperthyroidism. A 1-cm nodule was identified at the left upper pole of the thyroid. Fine-needle aspiration biopsy of this nodule revealed papillary carcinoma of the thyroid. Total thyroidectomy and a modified radical neck dissection were performed. Microscopic examination showed that the entire gland was replaced by a papillary carcinoma with a predominantly follicular pattern of growth. Of 44 lymph nodes tested, 31 were positive for metastatic papillary thyroid carcinoma. The presence of thyroid antibodies may have had a role in the aggressiveness of the patient's disease. CONCLUSION: To our knowledge, this is the first published report of a papillary thyroid cancer with manifestations resembling Graves' disease. PMID- 11421556 TI - Orbital lymphoma misdiagnosed as Graves' ophthalmopathy. AB - OBJECTIVE: To describe a case of bilateral orbital lymphoma mistakenly diagnosed as Graves' ophthalmopathy. METHODS: We present a case report, with laboratory data and photographic documentation, and discuss the differential diagnosis in patients with orbital masses. RESULTS: A 65-year-old man with bilateral exophthalmos and substantial weight loss was referred to the Endocrine Clinic for evaluation of possible Graves' disease. A 6-cm mass was detected in the left axilla. Biopsy of this mass revealed the histopathologic diagnosis of anaplastic B-cell lymphoma. Treatment with intrathecally administered methotrexate and orally administered dexamethasone promptly resulted in decreased proptosis. CONCLUSION: The most frequent cause of bilateral proptosis is Graves' ophthalmopathy, and when it is associated with weight loss in an elderly patient, the initial diagnostic consideration is thyrotoxic Graves' disease. This case should remind physicians that bilateral orbital lymphoma, although uncommon, may mimic Graves' ophthalmopathy. PMID- 11421557 TI - Selective estrogen receptor modulators: from bench to bedside and back. AB - OBJECTIVE: To provide a brief review of the history of the development of selective estrogen receptor modulators (SERMs), the current data assessing the effect of SERMs at the organ level, and the mechanism of action of these agents. METHODS: All the pertinent medical literature was reviewed, and the effects of SERMs on various end-organs were summarized. RESULTS: SERMs have been available for clinical use since the late 1960s. By the late 1980s, several SERMs had become available that influenced clinical practice. Multiorgan effects of these compounds include variable clinical efficacy for treatment of menopausal symptoms involving the central nervous system, variable effects on the genitourinary tract, and, in general, positive effects on serum lipid levels. SERMs seem to affect bone density positively, albeit to variable degrees, depending on the agent being used. The greatest effect of SERMs has been on the breast, and current SERMs seem to have efficacy for prevention of breast cancer as opposed to the controversial effect of estrogen on the breast. Disadvantages of SERMs include exacerbation of menopausal symptoms and, as with estrogen, an increased incidence of venous thrombosis and pulmonary emboli. SERMs act by modifying the configuration of the estrogen receptor. Effects at the gene transcription level seem to be tissue specific, a factor that likely accounts for the variability of clinical action seen. CONCLUSION: SERMs are a viable option for treatment of various problems associated with menopause. PMID- 11421558 TI - Tissue triglyceride levels in type 2 diabetes and the role of thiazolidinediones in reversing the effects of tissue hypertriglyceridemia: review of the evidence in animals and humans. PMID- 11421559 TI - Histologic variants of papillary thyroid carcinoma. PMID- 11421560 TI - Rapid availability of hemoglobin a1c value in the care of patients with diabetes. PMID- 11421561 TI - Loss of heterogeneous ribonucleoprotein A(2)/B(1) expression in thyroid neoplasms. AB - OBJECTIVE: To determine whether a difference exists in the immunohistochemical expression of heterogeneous ribonucleoprotein (hnRNP) A(2)/B(1) between benign and malignant thyroid lesions and to assess whether a gradient of expression could be found in normal thyroid, adenomas, and thyroid malignant tumors. METHODS: Formalin-fixed, paraffin-embedded archival tissues from 32 cases (8 nodular goiters, 8 follicular adenomas, 8 follicular carcinomas, and 8 papillary carcinomas) were immunostained with monoclonal antibody 703D4, directed against hnRNP A(2)/B(1), applied at a concentration of 10 microg/mL. The streptavidin biotin peroxidase method was used to label bound monoclonal antibody. Positivity was independently scored by two pathologists, who used a three-tiered scale. RESULTS: The benign thyroid tissues, including the hyperplastic and adenomatous lesions, demonstrated 3+ granular cytoplasmic staining for hnRNP A(2)/B(1), except in two cases (one nodular goiter and one follicular adenoma) in which 2+ staining intensity was noted. In contrast, the papillary and follicular carcinomas failed to stain with the antibody, except in two cases that showed weak (1+) staining. CONCLUSION: hnRNP A(2)/B(1) immunostaining appears to distinguish benign from malignant thyroid lesions. Loss of hnRNP A(2)/B(1) expression seems to be a characteristic feature of thyroid malignant lesions. PMID- 11421562 TI - Comparison of effects of thiazolidinediones on cardiovascular risk factors: observations from a clinical practice. AB - OBJECTIVE: To compare short-term glycosylated hemoglobin (HbA(1c)), lipid, weight, tolerability, and hepatic effects after switching patients with type 2 diabetes from troglitazone to either pioglitazone or rosiglitazone treatment. METHODS: This study compared the effects of conversion from maintenance troglitazone therapy to pioglitazone versus rosiglitazone. HbA(1c), lipids, weights, adverse effects, and hepatic status were monitored, providing no other major therapeutic change had been made. Of 163 study candidates, 144 and 125 patients fulfilled the criteria for comparison of HbA(1c) and lipids, respectively. RESULTS: HbA(1c) decreased an absolute mean of 0.08% for each treatment group, after a mean 3.2-month observation. Mean cholesterol, triglyceride, and low-density lipoprotein (LDL) cholesterol levels decreased in the pioglitazone group by 4.7%, 11.3%, and 7.3% but increased 8.4%, 38.4%, and 8.1%, respectively, in the rosiglitazone group. Mean high-density lipoprotein (HDL) increased 2.6% with pioglitazone and decreased 6.3% with rosiglitazone therapy. Patients receiving a statin concomitantly when switched to rosiglitazone treatment had a 51.9% mean triglyceride increase versus a 25.7% increase for those not taking a statin, whereas the patients switched to pioglitazone therapy had respective decreases of 14.2% and 6.2%. Both drugs were generally well tolerated; patients in both groups had similar slight weight increases and no hepatic dysfunction. CONCLUSION: Patients switched from maintenance troglitazone treatment to either pioglitazone or rosiglitazone therapy had similar glycemic control. Conversion to pioglitazone therapy caused a trend toward improvement in all lipid variables, but switching to rosiglitazone therapy caused significantly increased levels of cholesterol, triglycerides, and LDL and a trend toward decreased HDL. Patients already receiving statins when switched to rosiglitazone therapy had particularly notable triglyceride worsening. Whether these effects will lead to changes in cardiovascular outcome or will be maintained over a longer period remains to be established. PMID- 11421563 TI - Longitudinal assessment of economic burden and clinical outcomes in acromegaly. AB - OBJECTIVE: To determine costs and outcomes over time for surgical and various medical regimens in the management of patients with acromegaly. METHODS: We studied a sample of 53 consecutive Canadian patients with acromegaly who underwent a transsphenoidal pituitary surgical procedure only (N = 27) or in conjunction with medical therapy (N = 26). Outcomes were analyzed as person months spent in various health state outcomes, which were defined on the basis of growth hormone and insulin-like growth factor I levels. Costs are reported in 1998 Canadian dollars. RESULTS: The mean duration of follow-up was 49 months. Of the 53 patients, 25 (47%) had microadenomas at admission. Patients spent as much as 65% of the time in "uncured" health states. Patients with less extensive disease had better outcomes. The mean annual cost per patient was $8,111 (95% confidence interval, $5,848 to $10,374). Medications were the largest contributor to overall cost (38%). Although per patient surgical costs themselves were high (ranging from approximately $2,800 to $9,200), when averaged over the 4 years the mean annual cost was approximately $2,400, less than the cost of medications. Treatment of macroadenomas cost more than treatment of microadenomas ($11,425 versus $4,442 annually). The treatment of acromegaly costs $14.7 million annually in Canada (95% confidence interval, $10.6 to $18.8 million) and, if patterns of care are similar, about $139 million annually in the United States. CONCLUSION: Treatment of acromegaly is no more costly than therapy for other chronic diseases, especially those with a surgical component. Early diagnosis (at the stage of microadenoma) resulted in better outcomes and lower costs. Thus, from the standpoint of economics and well-being, a continued aggressive attitude toward screening programs and treatment of persistently active acromegaly seems warranted. PMID- 11421564 TI - Anticardiolipin antibodies in Hashimoto's disease. AB - OBJECTIVE: To ascertain the presence and significance of anticardiolipin antibodies in patients with Hashimoto's disease. METHODS: We studied 19 patients (15 women and 4 men; age range, 26 to 79 years) with Hashimoto's disease. IgA, IgG, and IgM anticardiolipin antibodies were measured by enzyme-linked immunosorbent assay. RESULTS: Of the 19 study patients, 4 (21%) tested positive for anticardiolipin antibodies (IgG in 2 and IgM in 2). Patients with anticardiolipin antibodies did not exhibit a higher titer of antithyroglobulin or anti-thyroid peroxidase antibodies in comparison with patients without anticardiolipin antibodies. Moreover, no relationship was found between anticardiolipin antibody levels and thyroid hormone replacement therapy. None of the patients with high levels of IgG or IgM anticardiolipin antibodies had clinical features of the antiphospholipid syndrome. CONCLUSION: Our results showed an increased incidence of anticardiolipin antibodies in patients with Hashimoto's disease. The presence of these antibodies may be a nonspecific marker of activation of the immune system. PMID- 11421565 TI - Liver involvement and abnormal iron variables in undiagnosed Addison's disease. AB - OBJECTIVE: To describe a case of untreated Addison's disease manifesting as severe gastrointestinal symptoms, persistently increased liver enzymes, substantially increased ferritin, and hepatic iron deposition and to document changes in these variables after corticosteroid replacement. METHODS: We thoroughly reviewed the clinical history and results of laboratory tests before and after treatment in a 23-year-old man during a period of 18 months. The relevant medical literature was also reviewed. RESULTS: The study patient had frequent episodes of severe abdominal symptoms, hemodynamic instability, and electrolyte imbalance. He underwent extensive laboratory investigations and was prescribed various treatment regimens. Increased levels of serum transaminases, ferritin, and transferrin saturation led to a liver biopsy, which showed lymphocytic infiltration and increased iron deposition. Eventually, cosyntropin stimulation (250 microg) confirmed the presence of adrenal insufficiency, and these abnormalities resolved after institution of daily administration of glucocorticoids and mineralocorticoids. CONCLUSION: Addison's disease can be a cause of unexplained hypertransaminasemia and profoundly increased ferritin levels. These changes are reversible but may lead to diagnostic confusion and delay in commencement of lifesaving corticosteroid therapy. PMID- 11421566 TI - Parathyromatosis: a cause for recurrent hyperparathyroidism. AB - OBJECTIVE: To report a case of parathyromatosis as a cause for recurrent hyperparathyroidism. METHODS: We present the case history, laboratory results, operative interventions, and pathologic findings in a 36-year-old woman. Relevant reports from the literature are reviewed. RESULTS: Our patient, who had been undergoing long-term hemodialysis because of renal failure, presented with secondary hyperparathyroidism and progressive bone pain. After an uneventful subtotal parathyroidectomy (removal of 3-1/2 glands), her symptoms resolved in conjunction with normalization of parathyroid hormone levels. Subsequently, however, recurrent hyperparathyroidism and severe bone pain necessitated second and third neck explorations, during which parathyromatosis was discovered. A total thyroidectomy was performed because of the bilateral nature of the disease. Postoperatively, the patient's bone pain resolved substantially, although her parathyroid hormone levels remained high. CONCLUSION: Parathyromatosis is a rare cause of recurrent hyperparathyroidism after parathyroidectomy. It consists of hyperfunctioning parathyroid tissues scattered throughout the neck, due either to intraoperative tissue spillage and subsequent implantation or to hyperplasia of parathyroid rests from embryologic development. This is one of the few case reports of parathyromatosis and the first case report of a mixed form of the disease, consisting of features of both subcapsular parathyroid rests and extracapsular implantation. PMID- 11421567 TI - Use of soy protein supplement and resultant need for increased dose of levothyroxine. AB - OBJECTIVE: To report a case of difficulty in achieving suppressive serum levels of thyroid hormone because of malabsorption of exogenous levothyroxine attributable to daily ingestion in close temporal relationship to the intake of a soy protein-containing food supplement. METHODS: We present the relevant history and laboratory data of the current case and provide supportive documentation from the literature. RESULTS: A 45-year-old woman who had hypothyroidism after a near total thyroidectomy and radioactive iodine ablative therapy for papillary carcinoma of the thyroid required unusually high oral doses of levothyroxine to achieve suppressive serum levels of free thyroxine (T(4)) and thyrotropin (thyroid-stimulating hormone or TSH). She had routinely been taking a "soy cocktail" protein supplement immediately after her levothyroxine. Temporal separation of the intake of the soy protein cocktail from the administration of the levothyroxine resulted in attainment of suppressive serum levels of free T(4) and TSH with use of lower doses of levothyroxine. CONCLUSION: Administration of levothyroxine concurrently with a soy protein dietary supplement results in decreased absorption of levothyroxine and the need for higher oral doses of levothyroxine to attain therapeutic serum thyroid hormone levels. PMID- 11421568 TI - Practical application of recombinant thyrotropin testing in clinical practice. AB - OBJECTIVE: To review the indications for use of recombinant thyrotropin (rTSH) and outline the details of implementation of rTSH diagnostic testing in patients with treated thyroid cancer. METHODS: We discuss the results of published clinical trials that have compared rTSH-stimulated testing with conventional withdrawal of thyroid hormone suppressive therapy. Appropriate candidates for rTSH testing are described, and the typical schedule for rTSH testing and follow up is presented. An overview of coding and documentation for reimbursement is also provided. RESULTS: Clinical studies have found no significant difference in the combined sensitivity of (131)I scans and serum thyroglobulin measurements for detection of recurrent thyroid cancer after rTSH stimulation versus withdrawal of thyroid hormone therapy. As expected, patients have fewer symptoms and a more favorable mood state after use of rTSH. Patients with thyroid cancer who have undergone total or near-total thyroidectomy followed by 131I ablation can be considered for rTSH testing. For low-risk patients, two cycles of rTSH testing 1 to 2 years apart, followed by testing every 3 to 5 years, are recommended. For moderate- to high-risk patients who have undergone one cycle of negative levothyroxine-withdrawal testing, two cycles of rTSH testing at a 6- to 12-month interval, followed by testing every 1 to 3 years for at least the first decade of follow-up, are recommended. Most commercial insurance, Medicare, and Medicaid carriers now cover rTSH, either in a prescription drug plan or under major medical benefits. CONCLUSION: Radioiodine scanning and serum thyroglobulin measurement after intramuscular injection of rTSH are valuable new monitoring options in patients with treated thyroid cancer, avoiding the adverse effects of hypothyroidism. PMID- 11421569 TI - Hope or hype? PMID- 11421570 TI - Potential cardiovascular effects of thiazolidinediones. PMID- 11421571 TI - Effect of risperidone dose on serum prolactin level. PMID- 11421572 TI - Endoscopic pelvic osteotomy for the treatment of hip dysplasia. AB - Adolescent and adult hip dysplasia can be surgically treated by rotating the acetabulum into a better weight-supporting position; however, open pelvic osteotomies are among the most invasive of all pediatric orthopaedic procedures. Endoscopic pelvic osteotomy offers the theoretical advantages of magnified visualization of the bone cuts, minimized surgical dissection, and rapid postoperative recovery. The technique of endoscopically assisted triple innominate osteotomy requires the combination of endoscopic skills and facility with more standard surgical approaches. PMID- 11421573 TI - Bone regeneration: new findings and potential clinical applications. AB - Bone is a biologically privileged tissue in that it has the capacity to undergo regeneration as part of a repair process. Fracture healing is the most common and recognizable form of bone regeneration, but several other examples of bone regeneration have been observed in humans, suggesting that the ability to regulate bone regeneration as a therapeutic tool should be possible. Historically, efforts at limb lengthening have led to procedures for regenerating bone, such as the method of Ilizarov. This procedure, known as distraction osteogenesis, has applications in a variety of skeletal conditions, including the restoration of large skeletal defects, the transport of bone in cases of severe trauma with bone loss, and the correction of skeletal deformities. Fibrodysplasia ossificans progressiva is an example of how an abnormal metabolic condition can be viewed as evidence for the capacity of humans to regenerate large amounts of bone if the cellular and molecular signaling events are altered. Elucidation of the cellular and molecular basis for bone regeneration in humans - particularly the role of the human genome in relation to the expression of various growth factors and cytokines, such as the bone morphogenetic proteins - offers great potential for the treatment of orthopaedic conditions. Development of specific bone morphogenetic proteins as therapeutic substances to induce bone regeneration in patients is well under way. As methods for enhancing fracture healing, distraction osteogenesis, and other procedures are refined, the development of protein- and gene-based therapies for regulating bone formation should lead to a new era of orthopaedic practice. PMID- 11421574 TI - Acute hematogenous osteomyelitis in children. AB - Acute hematogenous osteomyelitis in children is a relatively uncommon but potentially serious disease. Improvements in radiologic imaging, most notably magnetic resonance imaging, and a heightened awareness of this condition have led to earlier detection and resultant marked decreases in morbidity and mortality. Staphylococcus aureus, which has the ability to bind to cartilage, produce a protective glycocalyx, and stimulate the release of endotoxins, accounts for 90% of infections in all age groups. Infections with Haemophilus influenzae have become rare in immunized children. A careful history and a thorough physical examination remain important. Positive cultures are obtained in only 50% to 80% of cases; the yield is improved by the use of blood cultures and evolving molecular techniques. Improvements in antibiotic treatment have lessened the role of surgery in managing these infections. Sequential intravenous and high-dose oral antibiotic therapy is now an accepted modality. Evaluation of response to treatment by monitoring C-reactive protein levels has decreased the average duration of therapy to 3 to 4 weeks with few relapses. The emergence of antibiotic resistance, particularly resistance to methicillin and vancomycin by S aureus organisms, is of increasing concern. Long-term sequelae and morbidity are primarily due to delays in diagnosis and inadequate treatment. PMID- 11421575 TI - Nonarticular proximal tibia fractures: treatment options and decision making. AB - Nonarticular proximal-third fractures account for 5% to 11% of tibial shaft injuries and occur as a result of a variety of mechanisms. Treatment is more challenging than for more distal fractures, and the rates of compartment syndrome and arterial injury are higher, especially for displaced fractures. Closed management often leads to varus malunion, especially when the fibula is intact. Closed treatment should therefore be reserved for nondisplaced or minimally displaced fractures with little soft-tissue injury. Plating of the proximal tibia has become a less popular alternative because of the high incidence of infection and fixation failure. However, judicious use of lateral plates as an adjunct to medial external fixation in comminuted fractures can be effective. External fixation remains the most versatile method. It is indicated for fractures with short proximal fragments and in cases of extensive soft-tissue injury that would preclude use of other surgical techniques. Temporary joint-spanning external fixation has a role in the initial management of certain fracture patterns, particularly when accompanied by severe soft-tissue injury. Although intramedullary nailing can lead to valgus malunion in a sizable percentage of patients with this injury, it can be useful for stabilizing fractures with proximal fragments longer than 5 to 6 cm. Placing the entry portal more proximal and lateral, locking in extension, and using specific techniques, such as blocking screws, can improve alignment after nailing. Use of an algorithm that takes into account the severity of soft-tissue injury, the length of the fracture fragment, and the degree of fracture stability allows effective decision making among current treatment techniques. PMID- 11421576 TI - Magnetic resonance imaging of the foot and ankle. AB - Magnetic resonance (MR) imaging of the foot and ankle is playing an increasingly important role in the diagnosis of a wide range of foot and ankle abnormalities, as well as in planning for their surgical treatment. For an optimal MR study of the foot and ankle, it is necessary to obtain high-resolution, small-field-of view images using a variety of pulse sequences. The most common indication for MR imaging of the foot and ankle is for the evaluation of tendon and bone abnormalities, such as osteomyelitis, occult fractures, and partial and complete tears of the Achilles, tibialis posterior, and peroneal tendons. Magnetic resonance imaging has also been shown to be helpful in the diagnosis of several soft-tissue abnormalities that are unique to the foot and ankle, such as plantar fasciitis, plantar fibromatosis, interdigital neuromas, and tarsal tunnel syndrome. PMID- 11421577 TI - Wrist arthroscopy: principles and clinical applications. AB - With the development of better and smaller equipment, arthroscopy of the wrist offers the same benefits achievable with arthroscopy of the knee, shoulder, or elbow - not only diagnostic information but also a therapeutic option. Standardized techniques of performing wrist arthroscopy have been developed to evaluate the treat various wrist disorders, such as lesions of the triangular fibrocartilage complex, intra-articular distal radius fractures, and scaphoid fractures. Arthroscopy is now performed in the treatment of dorsal-wrist ganglion cysts and interosseous ligament disruptions, as well as for bone incisions, such as radial styloidectomy, distal ulnar excision (wafer procedure), and proximal row carpectomy. Compared with other techniques, arthroscopic procedures, such as repair of the triangular fibrocartilage complex, demonstrate better results and improved localization of the injury with a low complication rate. In addition, arthroscopic procedures involve lesssurgical dissection, less postoperative pain, a shorter recovery time, and an earlier return to work for the patient. PMID- 11421578 TI - Bone-graft harvesting from iliac and fibular donor sites: techniques and complications. AB - The ilium and the fibula are the most common sites for bone-graft harvesting. The different methods for harvesting iliac bone graft include curettage, trapdoor or splitting techniques for cancellous bone, and the subcrestal-window technique for bicortical graft. A tricortical graft from the anterior ilium should be taken at least 3 cm posterior to the anterior superior iliac spine (ASIS). Iliac donor site complications include pain, neurovascular injury, avulsion fractures of the ASIS, hematoma, infection, herniation of abdominal contents, gait disturbance, cosmetic deformity, violation of the sacroiliac joint, and ureteral injury. The neurovascular structures at risk for injury during iliac bone-graft harvesting include the lateral femoral cutaneous, iliohypogastric, and ilioinguinal nerves anteriorly and the superior cluneal nerves and superior gluteal neurovascular bundle posteriorly. Violation of the sacroiliac joint can be avoided by limiting the harvested area to 4 cm from the posterior superior iliac spine (PSIS) and by not penetrating the inner cortex. The caudal limit for bone harvesting should be the inferior margin of the roughened area anterior to the PSIS on the outer table to keep from injuring the superior gluteal artery. Potential complications of fibular graft harvesting include neurovascular injury, compartment syndrome, extensor hallucis longus weakness, and ankle instability. The neurovascular structures at risk for injury during fibular bone-graft harvesting include the peroneal nerves and their muscular branches in the proximal third of the fibular shaft and the peroneal vessels in the middle third. PMID- 11421579 TI - Metallothionein treatment reduces proinflammatory cytokines IL-6 and TNF-alpha and apoptotic cell death during experimental autoimmune encephalomyelitis (EAE). AB - Experimental autoimmune encephalomyelitis (EAE) is an animal model for the human autoimmune disease multiple sclerosis (MS). Proinflammatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) are considered important for induction and pathogenesis of EAE/MS disease, which is characterized by significant inflammation and neuroglial damage. We have recently shown that the exogenous administration of the antioxidant protein zinc metallothionein-II (Zn-MT-II) significantly decreased the clinical symptoms, mortality, and leukocyte infiltration of the CNS during EAE. However, it is not known how EAE progression is regulated nor how cytokine production and cell death can be reduced. We herewith demonstrate that treatment with Zn-MT-II significantly decreased the CNS expression of IL-6 and TNF-alpha during EAE. Zn MT-II treatment could also significantly reduce apoptotic cell death of neurons and oligodendrocytes during EAE, as judged by using TUNEL and immunoreactivity for cytochrome c and caspases 1 and 3. In contrast, the number of apoptotic lymphocytes and macrophages was less affected by Zn-MT-II treatment. The Zn-MT-II induced decrease in proinflammatory cytokines and apoptosis during EAE could contribute to the reported diminution of clinical symptoms and mortality in EAE immunized rats receiving Zn-MT-II treatment. Our results demonstrate that MT-II reduces the CNS expression of proinflammatory cytokines and the number of apoptotic neurons during EAE in vivo and that MT-II might be a potentially useful factor for treatment of EAE/MS. PMID- 11421580 TI - Role of apolipoprotein E receptors in regulating the differential in vivo neurotrophic effects of apolipoprotein E. AB - Apolipoprotein E (apoE) is known to bind to at least five receptors, including the low-density lipoprotein (LDL) receptor-related protein (LRP), very low density LDL receptor (VLDL-R), LDL-R, apoE receptor 2 (apoER2), and megalin/gp330. In this context, the main objective of the present study was to better understand the contributions of LRP and LDL-R to the in vivo neurotrophic effects of apoE. For this purpose, apoE-deficient and receptor-associated protein (RAP)-deficient mice were infused with recombinant apoE3, RAP, or saline. Infusion of apoE3 into apoE-deficient mice resulted in amelioration of degenerative alterations of pyramidal neurons, but had no effect on somatostatin producing interneurons. In contrast, infusion of apoE3 into RAP-deficient mice resulted in amelioration of degenerative alterations of somatostatin-producing interneurons. LRP and LDL-R levels were significantly reduced in RAP-deficient mice, but significantly increased in the apoE-deficient mice. In contrast, levels of apoE were reduced in the RAP-deficient mice compared to wildtype controls, suggesting that neurotrophic effects of apoE3 in the RAP-deficient mice were related to a combined deficit in endogenous apoE and selected apoE receptors. Furthermore, in apoE-deficient mice, infusion of apoE3 had a neurotrophic effect on somatostatin-producing interneurons only when combined with RAP, suggesting that increased expression of apoE receptors in apoE-deficient mice prevented apoE from rescuing somatostatin-producing neurons. This study supports the contention that some of the in vivo neurotrophic effects of apoE are mediated by LRP and LDL R and that a critical balance between levels of apoE and its receptors is necessary for the differential neurotrophic effects to appear. PMID- 11421581 TI - Calpain activity in the rat brain after transient forebrain ischemia. AB - Activity of the Ca(2+)-dependent protease calpain is increased in neurons after global and focal brain ischemia, and may contribute to postischemic injury cascades. Understanding the time course and location of calpain activity in the post-ischemic brain is essential to establishing causality and optimizing therapeutic interventions. This study examined the temporal and spatial characteristics of brain calpain activity after transient forebrain ischemia (TFI) in rats. Male Long Evans rats underwent 10 min of normothermic TFI induced by bilateral carotid occlusion with hypovolemic hypotension (MABP 30 mm Hg). Brain calpain activity was examined between 1 and 72 h after reperfusion. Western blot analysis of regional brain homogenates demonstrated a bimodal pattern of calpain-mediated alpha-spectrin degradation in the hippocampus, cortex, and striatum with an initial increase at 1 h followed by a more prominent secondary increase at 36 h after reperfusion. Immunohistochemical analysis revealed that calpain activity was primarily localized to dendritic fields of selectively vulnerable neurons at one hour after reperfusion. Between 24 and 48 h after reperfusion neuronal calpain activity progressed from the dorsal to ventral striatum, medial to lateral CA1 hippocampus, and centripetally expanded from watershed foci in the cerebral cortex. This progression was associated with fragmentation of dendritic processes, calpain activation in the neuronal soma and subsequent neuronal degeneration. These observations demonstrate a clear association between calpain activation and subsequent delayed neuronal death and suggest broad therapeutic window for interventions aimed at preventing delayed intracellular Ca(2+) overload and pathologic calpain activation. PMID- 11421582 TI - Selective antibody-induced cholinergic cell and synapse loss produce sustained hippocampal and cortical hypometabolism with correlated cognitive deficits. AB - The physiological interrelationships between cognitive impairments, neurotransmitter loss, amyloid processing and energy metabolism changes in AD, cholinergic dementia and Down's syndrome are largely unknown to date. This report contains novel studies into the association between cognitive function and cerebral metabolism after long-term selective CNS cholinergic neuronal and synaptic loss in a rodent model. We measured local cerebral rates of glucose utilization ((14)C-2-deoxyglucose) throughout the brains of awake rats 4.5 months after bilateral intraventricular injections of a cholinotoxic antibody directed against the low-affinity NGF receptor (p75 NGF) associated with cholinergic neurons (192 IgG-saporin). Permanent cholinergic synapse loss was demonstrated by [(3)H]-vesamicol in vitro autoradiography defining presynaptic vesicular acetylcholine (ACh) transport sites. While other metabolic studies have defined acute and transient glucose use changes after relatively nonspecific lesions of anatomical regions containing cholinergic neurons, our results show sustained reductions in glucose utilization in brain regions impacted by cholinergic synapse loss, including frontal cortical and hippocampal regions, relative to glucose use levels in control rats. In the same animals, impaired cognitive spatial performance in a Morris water maze was correlated with reduced glucose use rates in the cortex and hippocampus at this time point, which is consistent with increased postmortem cortical and hippocampal amyloid precursor protein (APP) levels (45, 46). These results are consistent with the view of cholinergic influence over metabolism, APP processing, and cognition in the cortex and hippocampus. PMID- 11421583 TI - Isolation of multipotent neural precursors residing in the cortex of the adult human brain. AB - Multipotent precursors able to generate neurons, astrocytes, and oligodendrocytes have previously been isolated from human brain embryos and recently from neurogenic regions of the adult human brains. The isolation of multipotent neural precursors from adult human should open new perspectives to study adult neurogenesis and for brain repair. The present study describes the in vitro isolation from adult human brains of a progenitor responsive to both epidermal and basic fibroblast growth factors that forms spheres as it proliferates. Single spheres derived from various regions of the brain generate in vitro neurons, astrocytes, and oligodendrocytes. The clonal origin of the spheres was revealed by genomic viral insertion using lentiviral vector. Interestingly, this vector appears to be a potent tool for gene transfer into human neural progeny. Ninety six percent of the spheres investigated were multipotent. Multipotent precursors were isolated from all brain regions studied, including the temporal and the frontal cortex, the amygdala, the hippocampus, and the ventricular zone. This study is the first evidence that primitive precursors such as multipotent precursors exist in the adult human cortex and can reside far from the ventricles. Neurogenesis derived from adult human progenitors differ to murine neurogenesis by the requirement of laminin for oligodendrocyte generation and by the action of basic-fibroblast growth factor and platelet derived growth factor that prevented the formation of oligodendrocytes and neurons. Moreover, the differentiation of human adult precursors seems to differ from fetal ones: adult precursors do not necessitate the removal of mitogen for differentiation. These results indicate that the study of adult multipotent precursors is a new platform to study adult human neurogenesis, potentially generate neural cells for transplantation, and design protocols for in vivo stimulation. PMID- 11421584 TI - Phosphorylation of signal transducer and activator of transcription-3 (Stat3) after focal cerebral ischemia in rats. AB - JAK-STAT is the major downstream signal pathway of interleukin-6 (IL-6) cytokine family and is regulated by Tyr705 phosphorylation of Stat3. The present study examined the extent and the localization of phosphorylated Stat3 protein in brain tissue after focal ischemia in rats. The localizations of unphosphorylated and phosphorylated Stat3 were immunohistochemically examined in rats after 0.5 to 168 h of reperfusion following 1.5 h of middle cerebral artery occlusion (MCAO), induced by the intraluminal suture method. Absolute phosphorylated Stat3 immunoreactive cell counts were made in the cerebral cortex (ischemic core, peri ischemia region, and contralareral cortex) and lateral striatal regions on both the ischemic and the contralateral sides. Stat3 protein was localized diffusely in cortical and striatal neurons in the sham-operated animals. Although weak Stat3 staining was detected in damaged neurons in the ischemic region, activated microglia, astrocytes, and endothelial cells clearly expressed Stat3 in this region. On the other hand, the sham group showed no phosphorylated Stat3 immunoreactivity. Phosphorylated Stat3 immunoreactivity was first detected in neurons after 3.5 h of reperfusion in each cortical and striatal region. Thereafter, Stat3 phosphorylation was marked in neurons in the peri-infarct region, peaked at 24 h, and then gradually declined throughout the reperfusion period. Endothelial cells expressed phosphorylated Stat3 in the ischemic core at 48 h of reperfusion. To identify the cellular source of phosphorylated Stat3, lectin histochemical study and immunohistochemical study with anti-microtubule associated proten-2 and anti-glial fibrillary acidic protein antibodies were carried out. Double-staining immunohistochemistry with these cellular makers revealed phosphorylated Stat3 to be present in neurons, but in neither astrocytes nor microglia/macrophages. These results were also confirmed be western blot analysis. The present results indicate that Stat3 activation occurs in neurons and endothelial cells only during post-ischemic reperfusion despite widespread expression of IL-6 cytokines. PMID- 11421585 TI - The regrowth of axons within tissue defects in the CNS is promoted by implanted hydrogel matrices that contain BDNF and CNTF producing fibroblasts. AB - In this study we demonstrate the potential for combining biocompatible polymers with genetically engineered cells to elicit axon regrowth across tissue defects in the injured CNS. Eighteen- to 21-day-old rats received implants of poly N-(2 hydroxypropyl)-methacrylamide (HPMA) hydrogels containing RGD peptide sequences that had been infiltrated with control (untransfected) fibroblasts (n = 8), fibroblasts engineered to express brain-derived neurotrophic factor (BDNF) (n = 5), ciliary neurotrophic factor (CNTF) (n = 5), or a mixture of BDNF and CNTF expressing fibroblasts (n = 11). Fibroblasts were prelabeled with Hoechst 33342. Cell/polymer constructs were inserted into cavities made in the left optic tract, between thalamus and superior colliculus. After 4-8 weeks, retinal projections were analyzed by injecting right eyes with cholera toxin (B-subunit). Rats were perfused 24 h later and sections were immunoreacted to visualize retinal axons, other axons (RT97 antibody), host astrocytes and macrophages, donor fibroblasts, and extracellular matrix molecules. The volume fraction (VF) of each gel that was occupied by RT97(+) axons was quantified. RT-PCR confirmed expression of the transgenes prior to, and 5 weeks after, transplantation. Compared to control rats (mean VF = 0.02 +/- 0.01% SEM) there was increased ingrowth of RT97(+) axons into implants in CNTF (mean VF = 0.33 +/- 0.19%) and BDNF (mean VF = 0.62 +/-0.19%) groups. Axon growth into hydrogels in the mixed BDNF/CNTF group (mean VF = 3.58 +/- 0.92%) was significantly greater (P < 0.05) than in the BDNF or CNTF fibroblast groups. Retinal axons exhibited a complex branching pattern within gels containing BDNF or BDNF/CNTF fibroblasts; however, they regrew the greatest distances within implants containing both BDNF and CNTF expressing cells. PMID- 11421586 TI - Pegylated brain-derived neurotrophic factor shows improved distribution into the spinal cord and stimulates locomotor activity and morphological changes after injury. AB - The neurotrophin brain-derived neurotrophic factor (BDNF) shows promise for the treatment of central nervous system (CNS) trauma and disease. Effective delivery methods are required, however, for BDNF to be useful as a therapeutic agent. To this end, we examined the penetration of intrathecally infused N-terminal pegylated BDNF (peg-BDNF) compared to similar infusion of native BDNF after spinal cord injury (SCI). Pegylation dramatically improved delivery of BDNF to the spinal cord and induced the expression of Fos in spinal cord neurons. To test whether enhanced delivery would improve the modest effects on behavioral recovery and axonal outgrowth observed with native BDNF infusion, we assessed the efficacy of 2-week 25 microg/day peg-BDNF treatment, beginning 12-24 h (early) or 15 days (delayed) after midthoracic spinal contusion. Similar to native BDNF, early treatment with peg-BDNF accelerated the recovery of stepping in the open-field and acutely stimulated locomotor central pattern generator activity, as seen by the activation of hindlimb airstepping during either period of administration. The infusion of peg-BDNF, regardless of the timing of delivery, was related to enhanced sprouting of putative cholinergic fibers, like that observed after high dose native BDNF treatment. Despite improved delivery, however, neither axonal responses nor the extent of locomotor recovery were enhanced compared to native BDNF treatment. This suggests that alternative strategies, such as neurotrophin treatment in conjunction with cell transplantation techniques, or treatment nearer the cell bodies of target neurons might be employed in an attempt to effect significant repair after SCI. PMID- 11421587 TI - Short-term estrogen replacement increases beta-preprotachykinin mRNA levels in uninjured dorsal root ganglion neurons, but not in axotomized neurons. AB - Dorsal root ganglion (DRG) neurons that mediate nociception express the high affinity NGF receptor (trkA) gene and the preprotachykinin (PPT) gene. NGF has been shown to regulate both of these DRG neuronal genes. Our laboratory has shown that these genes are also regulated by estrogen. Long-term daily estrogen replacement, in adult ovariectomized (OVX) rats, causes a coordinate decline in trkA and beta-PPT mRNA levels in lumbar DRG neurons, while short-term estrogen replacement increases trkA mRNA levels in uninjured as well as in axotomized lumbar DRG neurons. The purpose of the current study was to test the hypothesis that short-term estrogen replacement increases DRG beta-PPT mRNA levels in lumbar DRG neurons of OVX rats and that the increase is dependent on target-derived NGF. Sciatic nerve transection (SNT) was used to eliminate target-derived NGF in L4 and L5 DRGs in adult OVX rats. Seven days later, one-half of the SNT and one-half of the animals that had received sham sciatic nerve transactions (SHAM) received two daily injections of estradiol benzoate (EB). The remaining rats received two daily injections of vehicle alone. Quantitative in situ hybridization analyses of sections from L4 and L5 DRGs showed that two daily injections of EB significantly increased beta-PPT mRNA levels in DRGs of SHAM animals, but had no effect on beta PPT mRNA levels in DRGs from SNT animals. These data coupled with our earlier observations of the effect of short-term estrogen replacement on DRG trkA mRNA levels, indicate that the regulation of DRG beta-PPT mRNA levels by estrogen requires target-derived NGF. PMID- 11421588 TI - Spatiotemporal analysis of Fos expression associated with cocaine- and PTZ induced seizures in prenatally cocaine-treated rats. AB - We previously reported that prenatal cocaine exposure (40 mg/kg s.c., E10-E20) increased susceptibility to convulsant-induced seizures later in life, with female rats becoming more sensitive to seizures induced by cocaine and pentylenetetrazol (PTZ), and males more sensitive to PTZ-induced seizures (Snyder Keller and Keller, 1995, 2000). In order to determine the locus of enhanced seizure susceptibility in the brains of prenatally cocaine-treated rats, we examined the distribution and density of Fos-immunoreactive cells after cocaine- and PTZ-induced seizures in mature rats. Subconvulsive cocaine doses induced c fos in cortical areas as well as densely dopamine-innervated regions such as striatum and nucleus accumbens. Following cocaine-induced seizures, intense c-fos induction was observed in piriform cortex, amygdala, and hippocampus. Quantification of the number of Fos-immunoreactive cells in the brains of prenatally cocaine-treated versus prenatally saline-treated rats revealed differences in piriform cortex and amygdala that were indicative of a lower threshold in prenatally cocaine-treated female rats. Following PTZ-induced seizures, the same pattern of limbic structures were recruited with increasing seizure severity. Only females exhibited changes in the number of Fos immunoreactive cells as a result of prenatal cocaine treatment. Pretreatment with the noncompetitive NMDA antagonist MK-801 blocked both cocaine- and PTZ-induced seizures, and Fos expression in limbic areas was also blocked. The dopamine D1 antagonist SCH 23390 blocked cocaine-induced seizures and associated c-fos induction, but not PTZ-induced seizures or Fos. Examination of the pattern of Fos expression at 15-20 min postseizure revealed that the initial site of c-fos induction associated with PTZ-induced seizures appeared to be the piriform cortex, whereas cocaine-induced seizures induced early expression in both piriform cortex and lateral amygdala. These findings suggest that neural alterations residing in the piriform cortex and amygdala are likely to account for the increased seizure susceptibility of prenatally cocaine-treated rats. PMID- 11421589 TI - Evaluation of nigrostriatal dopaminergic function in adult +/+ and +/- BDNF mutant mice. AB - Deletion of a single copy of the BDNF gene has been shown to affect the nigrostriatal dopaminergic system of young adult BDNF mice. In the present report we evaluated various indices of nigrostriatal dopaminergic function between 9 month-old wild-type (+/+) and heterozygous (+/-) BDNF mutant mice. Performance in a sensorimotor beam walking task was significantly decreased in +/- mice as indicated by increased times required to traverse both a wide (21 mm) and narrow (6 mm) beam. No differences in spontaneous locomotor behavior were observed between the +/+ and +/- mice. Amphetamine-stimulated (5 mg/kg) locomotor behavior was increased to a greater degree in the +/- mice, with the number of movements performed by these mice being significantly greater than their +/+ controls. Corpus striatal dopamine concentrations were significantly greater in the +/- BDNF mice. The absence of any significant differences for dopamine concentrations within the hypothalamus and olfactory bulb of these mice, as well as an absence of any difference in striatal norepinephrine concentrations, suggested a relative specificity of these effects to the corpus striatum. Both the +/- and +/+ mice showed similar reductions in striatal dopamine concentrations in response to a neurotoxic regimen of methamphetamine (20 mg/kg). Collectively these data show increased levels of striatal dopamine concentrations associated with altered behavioral responses involving the nigrostriatal dopaminergic system within the heterozygous BDNF mutant mice. PMID- 11421590 TI - Differential neuroprotection from human heat shock protein 70 overexpression in in vitro and in vivo models of ischemia and ischemia-like conditions. AB - We previously showed that overexpressing the 70-kDa inducible heat shock protein in primary astrocyte cultures and in a rodent stroke model using viral vectors resulted in protection from ischemia and ischemia-like injury. However, viral transfection could potentially provoke a stress response itself; therefore, we examined whether transgenic mice constitutively expressing human heat shock protein 70 were protected from ischemic insults. Astrocyte cultures from brains of heat shock protein 70 transgenic mice were resistant to hydrogen peroxide injury in a dose-dependent fashion, but were less resistant to hypoglycemia and oxygen-glucose deprivation. Because hydrogen peroxide exposure and glucose deprivation are partially dependent on glutathione levels, we determined whether heat shock protein 70 transgenic cultures had altered glutathione levels under normal growth conditions. However, there was no significant difference in glutathione levels between heat shock protein 70 transgenic and wildtype astrocytes. Hippocampal, but not cortical neuron cultures from these same transgenic mice were also protected against oxygen-glucose deprivation and glutamate toxicity. In an in vivo model of permanent focal cerebral ischemia, there was no significant difference in infarct size assessed 24 h postinsult. These results suggest that heat shock protein 70 protects against some but not all kinds of central nervous system injury. The protective effects may be related to the nature and severity of the insults, as well as subpopulations of brain cells and dose-dependent effects of HSP70 overexpression. PMID- 11421591 TI - Regional expression of Par-4 mRNA and protein after fluid percussion brain injury in the rat. AB - Regional levels of prostate apoptosis response-4 (Par-4) protein and mRNA were measured after lateral fluid percussion (FP) brain injury in rats. Immunochemical studies indicated that Par-4 immunoreactivity (ir) is present in cortical neurons and hippocampal CA1-CA3 pyramidal neurons in uninjured rats. Increases of Par-4 ir were observed in the CA3 neurons of the ipsilateral hippocampus (IH), but not in injured left cortex (IC) at 48 h after FP brain injury. Levels of the Par-4 mRNA measured by RT-PCR assay and protein measured by Western blot procedure were significantly increased in the injured IC and IH, but not in the contralateral right cortex and hippocampus after brain injury. Levels of both Par-4 protein and mRNA were significantly increased in the IC and IH as early as 2 h and stayed elevated at 24 and 48 h after injury. These data show that the induction of proapoptotic Par-4 mRNA and protein occurs only in the IC and IH that have been observed to undergo apoptosis and neuronal cell loss after lateral FP brain injury. Because increased expression of Par-4 has been observed to contribute to apoptosis and cell death in cultured neurons, the present temporal pattern of Par 4 expression is consistent with a role for Par-4 in apoptosis and neuronal cell death after traumatic brain injury. PMID- 11421592 TI - Streptozotocin-induced diabetes causes metabolic changes and alterations in neurotrophin content and retrograde transport in the cervical vagus nerve. AB - Abnormal availability of neurotrophins, such as nerve growth factor (NGF), has been implicated in diabetic somatosensory polyneuropathy. However, the involvement of neurotrophins in diabetic neuropathy of autonomic nerves, particularly the vagus nerve which plays a critical role in visceral afferent and in autonomic motor functions, is unknown. To assess the effects of hyperglycemia on the neurotrophin content and transport in this system, cervical vagus nerves of streptozotocin (STZ)-induced diabetic rats were studied at 8, 16, and 24 weeks after the induction of diabetes. Elevations in vagus nerve hexose (glucose and fructose) and polyol levels (sorbitol), and their normalization with insulin treatment, verified that the STZ treatment resulted in hyperglycemia-induced metabolic abnormalities in the nerve. Neurotrophin (NGF and neurotrophin-3; NT-3) content and axonal transport were assessed in the cervical vagus nerves from nondiabetic control rats, STZ-induced diabetic rats, and diabetic rats treated with insulin. The NGF, but not the NT-3, content of intact vagus nerves from diabetic rats was increased at 8 and 16 weeks (but not at 24 weeks). Using a double-ligation model to assess the transport of endogenous neurotrophins, the retrograde transport of both NGF and NT-3 was found to be significantly reduced in the cervical vagus nerve at later stages of diabetes (16 and 24 weeks). Anterograde transport of NGF or NT-3 was not apparent in the vagus nerve of diabetic or control rats. These data suggest that an increase in vagus nerve NGF is an early, but transient, response to the diabetic hyperglycemia and that a subsequent reduction in neuronal access to NGF and NT-3 secondary to decreased retrograde axonal transport may play a role in diabetes-induced damage to the vagus nerve. PMID- 11421593 TI - Neuroprotective effects of lamotrigine and remacemide on excitotoxicity induced by glutamate agonists in isolated chick retina. AB - The possible neuroprotective effects of two recently developed antiepileptic compounds, lamotrigine (LTG) and remacemide (REMA), against glutamate agonist induced excitotoxicity have been investigated in the isolated chick embryo retina model. Retina segments from 15- or 16-day-old embryos were incubated in 1 ml of balanced salt solution, at 25 degrees C for 30 min, in the presence or absence of N-methyl-d-aspartate (NMDA), kainic acid (KA), or alpha-amino-3-hydroxy-5-methyl 4-isoxazolepropionic acid (AMPA) (10 to 200 microM). LTG, REMA, and the active desglycinyl metabolite of REMA (d-REMA) (10-200 microM) were added separately 5 min before glutamate agonists. Retina damage was assessed after 24 h (i) by measuring LDH activity present in the medium, expressed as percentage of total retina LDH activity, and (ii) by histological analysis of retina specimens through scoring for the presence or absence of edema, necrosis, nuclear pyknosis, and cell layer damage. LTG, REMA, and d-REMA reduced LDH release produced by NMDA 58-70% in a dose-dependent manner, with d-REMA being the most potent (EC(50): d REMA, 25.75 +/- 3.27 microM; REMA, 64.75 +/- 7.75 microM; LTG, 60.50 +/- 6.80 microM; P < 0.001). The drugs had less effect on the LDH release produced by AMPA and KA. Histological analysis confirmed these biochemical results, with all three compounds reducing edema and the number of necrotic and pyknotic nuclei in the ganglion layer. d-REMA provided almost complete protection of the ganglion cell layer against damage produced by NMDA. Combinations of d-REMA and LTG showed additive rather than potentiative effects against NMDA-induced cell injury. The present data provide pharmacological evidence that LTG, REMA, and d-REMA decrease glutamate agonist-induced excitotoxicity in isolated chick retina, findings that might have therapeutic implications for various neurological disorders. PMID- 11421594 TI - N-methyl-D-aspartate receptor mRNA levels change during reproductive senescence in the hippocampus of female rats. AB - Estrogen interacts with N-methyl-d-aspartate (NMDA) receptors to regulate multiple aspects of morphological and functional plasticity. In the hippocampus, estrogens increase both dendritic spine density and synapse number, and NMDA antagonists block these effects. This plasticity in the hippocampus mediated by estrogen may be of particular importance in the context of aging when estrogen levels change and cognitive function is often impaired. Therefore, the present study was designed to investigate effects of aging and reproductive status on NMDA receptor (NR) subunit mRNA levels in the hippocampus. NR1, NR2A, and NR2B mRNA levels were measured by RNase protection assay in young (3-4 month), middle aged (12-13 month), and aged (24-25 month) Sprague-Dawley rats in different phases of the estrous cycle in cycling animals and in acyclic subjects. Our results demonstrated that NMDA receptor subunit mRNA levels were much more prominently affected by the chronological age than by the reproductive status of the animals. Age-related changes were observed in NR1, NR2A, and NR2B in the ventral hippocampus and in NR1 and NR2B in the dorsal hippocampus. However, the only relationship with reproductive status was seen for NR1 mRNA, and this was restricted to the ventral hippocampus. An interaction between chronological age and reproductive status was found, with higher levels of NR1 mRNA seen in young animals in proestrus than in those in diestrus I (high and low estrogen levels, respectively). However, this relationship was not seen in the aged subjects. These results demonstrate that the hippocampus is subjected to age-related alterations in NMDA receptor subunit mRNA levels and that animals of different ages are influenced differently by reproductive status. This shift in the NMDA receptor mRNA levels may be a possible molecular mechanism contributing to alterations in cognitive behavior during normal aging. PMID- 11421595 TI - Dimethyl sulfoxide suppresses NMDA- and AMPA-induced ion currents and calcium influx and protects against excitotoxic death in hippocampal neurons. AB - Dimethyl sulfoxide (DMSO) is widely used in neuroscience research as a solvent for various pharmacological agents in both cell culture and in vivo studies and is also used in humans to treat musculoskeletal problems and pain. We now report that concentrations of DMSO to which neurons are typically exposed in experimental studies and in human patients (0.5-1.5%) inhibit glutamate responses in hippocampal neurons. DMSO suppresses, in a rapidly reversible manner, electrophysiological responses and calcium influx induced by glutamate, N-methyl d-aspartate, and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate. Moreover, DMSO can prevent excitotoxic death of the neurons. These findings have important implications for the use of DMSO as a solvent in studies that involve glutamatergic neurotransmission. Our data also identify a mechanism that might explain clinical effects of DMSO on both peripheral and CNS neurons and suggest a potential use for DMSO in the treatment of excitotoxic neurodegenerative conditions. PMID- 11421596 TI - A novel approach for studying endogenous abeta processing using cultured primary neurons isolated from APP transgenic mice. AB - The central component of senile amyloid plaques in Alzheimer's disease (AD) is the beta-amyloid peptide (Abeta), derived from proteolytic processing of the amyloid precursor protein (APP). In this study, we developed an in vitro model to measure and identify soluble Abeta from primary cortical neurons. Neurons were isolated from mice transgenic for human APP695 containing the K670N, M671L double mutation. We characterized soluble Abeta using Western blot and ELISA assays. We found that the Abeta levels in conditioned media from these neurons were readily detectable and almost five times higher than in CSF. The majority of Abeta in the media was Abeta1-40; however, Abeta1-42 was also detectable. When the neurons were exposed to Phorbol 12-myristate 13-acetate (PMA), alpha1-antichymotrypsin, or alpha1-antitrypsin, the alterations of soluble Abeta levels were consistent with other models reported. Most importantly, the soluble Abeta in our model was remarkably stable, and aliquots were unchanged after prolonged incubations or repeated freeze/thaw cycles. The Abeta appeared to be monomeric by Western blot analysis. Soluble Abeta coimmunoprecipitated with endogenous mouse apolipoprotein E from the primary cultures. Taken together, our data demonstrated that using a Western blot assay to detect soluble Abeta from transgenic mouse overexpressing APP695 is sensitive, specific, and reliable and provides an accessible model for examining the neuronal metabolism of APP and Abeta. PMID- 11421597 TI - Increased ischemia-reperfusion blood flow impairs the skeletal muscle contractile function. AB - BACKGROUND: The ultimate aim of replantations and transplantations of skeletal muscle is to improve impaired function. The purpose of this study was to examine the contribution of varying durations of ischemia to postischemic blood flow in the skeletal muscle and the contribution of modulation of postischemic blood flow to skeletal muscle function and viability, using an ischemic revascularized hind limb model in rats. MATERIALS AND METHODS: Warm ischemia produced by vascular pedicle clamping was sustained for 90 min, 3 h, or 6 h. Postischemic blood flow was measured by a Doppler flowmeter or microsphere technique. In another series of experiments of 3-h ischemia, either saline or N(G)-methyl-l-arginine acetate (l-NMMA) was infused for the first 2 h of reperfusion. Postischemic blood flow was also measured. Muscle contractile function and viability were determined after 24 h of reperfusion. RESULTS: Postischemic blood flow was significantly increased during the first 10 min of reperfusion in the 90-minute ischemic group and during the first 2 h in the 3-h ischemic group compared with contralateral control blood flow. No significant increase in postischemic blood flow was noted in the 6-h ischemic group. Postischemic blood flow was significantly decreased by the l-NMMA infusion. Contractile function and viability of the tibialis anterior muscle and contractile function of the gastrocnemius muscle in the l-NMMA group were significantly increased. CONCLUSION: Reperfusion blood flow increased time dependently until 3 h of warm ischemia. Hyperemia deteriorated skeletal muscle contractile function, although it was well preserved by l-NMMA infusion to restrict the postischemic hyperemia. PMID- 11421598 TI - Perflubron emulsion reduces inflammation during extracorporeal circulation. AB - The recovery from cardiac surgery and cardiopulmonary bypass can be complicated by an acute inflammatory response. Circulating blood through an extracorporeal circuit (ECC) contributes to this complication. Perfluorocarbon-based blood substitutes (PFCs) are under investigation for use as a component of the ECC "prime" solution, because PFCs increase the oxygen-carrying capacity of the diluted blood. Some PFCs may provide the additional benefit of attenuating the ECC-induced inflammatory response. Earlier, we reported that perflubron emulsion (PFE, Alliance Pharmaceutical Corp.) reduced neutrophil (PMN) activation in vivo. However, the potential of PFE to reduce ECC-induced PMN activation has not been investigated. In this study, we used a small-scale ECC model to quantify the extent of PMN activation during circulation and to examine if PFE treatment attenuated PMN activation. ECC circuits were filled with a mixture of blood and Plasmalyte. Two groups were studied: an untreated group containing blood plus PlasmaLyte and a treated group in which some of the Plasmalyte was substituted with PFE (4.5 g/100 ml). Hematology and measures of whole blood PMN activation were made from blood samples taken periodically throughout the 120-min ECC circulation period. We found, for the untreated group, a significant decrease in the number of circulating PMNs and an increase in PMN activation with time. PMN activation was demonstrated as a significant increase in the expression of the PMN adhesion protein CD11b (P < 0.05) and an increase in PMN oxygen free radical production (reactive oxygen species (ROS)). After 120 min of circulation, the PMNs remained capable of a significant response to a second inflammatory stimulus, but PFE treatment significantly attenuated the fMLP-induced increase in PMN ROS at t = 120 min (P < 0.05). These results suggest that PFE may have dual utility in cardiac surgery, to increase oxygen delivery and to serve as an antiinflammatory agent. PMID- 11421599 TI - Contribution of serotonin to liver injury following canine small-intestinal ischemia and reperfusion. AB - Intestinal ischemia and reperfusion (I/R) has been shown to be associated with multiple organ damages. Serotonin (5-hydroxytriptamine; 5-HT), which is synthesized in the enterochromaffin cells in the intestine and stored in platelets, is known to play an important role in platelet aggregation and vasoconstriction and may ultimately enhance such organ injuries. The purpose of this study was to investigate the association between liver damage and 5-HT levels in the liver after intestinal I/R. The entire canine small intestine, isolated on a vascular pedicle that consisted of the proximal superior mesenteric artery and superior mesenteric vein, was subjected to 4-h ischemia by clamping these vessels and the marginal arteries supplying the proximal and distal ends of the small intestine. Hepatic blood flow, liver tissue blood flow, bile flow rate, and hepatic venous ketone body ratio (HVKBR) were measured before and at the end of intestinal ischemia and at 5, 15, and 30 min, and 1 and 2 h after reperfusion. 5-HT levels in plasma of the portal vein and hepatic vein were assayed at the same intervals. Time-matched, sham-operated animals served as controls. Intestinal I/R significantly decreased the liver tissue flow, bile flow rate, and HVKBR. Compared to those in controls, 5-HT levels in the portal vein and hepatic vein were markedly increased after reperfusion. Furthermore, intravenous administration of 5-HT receptor antagonists attenuated the liver dysfunction after intestinal reperfusion. These results suggest that intestinal I/R induces continuous disturbance of hepatic microcirculation, leading to liver dysfunction, and that 5-HT may be implicated as one of the mediators of liver dysfunction after intestinal I/R. PMID- 11421600 TI - Nitric oxide protects the intestine from the damage induced by laparotomy and gut manipulation. AB - BACKGROUND: The intestine is highly susceptible to free radical-induced damage, and our earlier work has shown that surgical stress induces the generation of oxygen free radicals in enterocytes, resulting in intestinal damage along with ultrastructural changes. Since nitric oxide (NO) is an important mediator of gastrointestinal function, this study looked at the effect of NO on surgical stress-induced intestinal alterations. MATERIALS AND METHODS: Control rats and rats pretreated with the NO donor l-arginine were subjected to surgical stress by opening the abdominal wall and handling the intestine as done during laparotomy. Enterocytes were isolated and homogenate prepared, and the protection offered by l-arginine against damage due to surgical stress was determined and compared with normal controls. Protection to structural as well as functional aspects of the intestine was also examined. RESULTS: Intestinal manipulation affected intestinal structure as assessed by electron microscopy. Functional impairment of the enterocyte was also evident, with increased xanthine oxidase activity resulting in production of superoxide anion. This impairment is more dramatic in the crypt cells. Increased protease activity was also seen following laparotomy and handling. Pretreatment with the NO synthase substrate l-arginine prevented these damaging effects. Arginine protection was abolished in the presence of the NO synthase inhibitor NG-nitro-l-arginine methyl ester, indicating the role of NO. CONCLUSION: Stress in the small intestine due to any surgery can affect enterocyte structure and function. These damaging effects can be prevented by NO, an important modulator of cellular function. PMID- 11421601 TI - Ventral incisional hernia recurrence. AB - During the period October 1993 to December 1996, 31 patients were operated on by the author for primary or recurrent ventral incisional hernia (VIH). Three patients were excluded from analysis because their records were unavailable for review. The median age of the 28 remaining patients at their initial procedure was 57.5 years (range, 37-78 years). The repair was performed with interrupted O Ethibond sutures in all but 3 cases where Prolene suture was used secondary to noniatrogenic contamination or recurrent hernia. There were no unplanned enterotomies in the entire series and prophylactic intravenous antibiotics were used in all cases. The only significant complications were skin hyperemia after five repairs in 3 patients who were treated empirically with intravenous antibiotics, and 1 patient who had an antibiotic-associated rash. There were no 30-day mortalities. Prolene mesh was used exclusively in all repairs performed with mesh. Seven of these repairs (25%) were for recurrent VIH. Three of these seven patients had previous mesh repairs. Six of these seven patients who presented with recurrent VIH had a mesh repair and four developed a recurrence. Five of seven were active smokers, with one having severe obstructive lung disease. Four of seven related significant occupational lifting. Of the 21 patients having initial repair of VIH, mesh was used in 8 (38%). After a median follow-up of 13 months, there were 2 recurrent hernias (25%). The remaining 13 patients had primary closure of their hernias. After median follow-up of 25 months, there were 5 recurrences (38%). A total of 34 VIH repairs were performed on these 28 patients, of which 13 were for recurrent hernias. Five of thirteen (38%) of the mesh repairs for recurrent VIH failed. The median body mass index (BMI) for the 13 patients having primary repair was 26.4, and that for all 21 cases having mesh repair was 28.8. Patients with recurrent VIH frequently recur despite use of mesh, avoidance of contamination, and consistent technique. No difference in BMI was apparent in those who recurred. Continued smoking and occupational lifting may be important risk factors for recurrent VIH. PMID- 11421602 TI - Blood flow rate and energy charge in the isolated perfused canine liver. AB - Background. Liver viability depends on blood flow rate, which varies among isolated perfused livers used in different studies. We aimed to identify the minimal blood flow capable of keeping the energy charge compatible with liver viability. Materials and methods. Isolated canine livers were assigned to different groups (n = 4) with blood flow rates of 0.5 (Group I), 1.0 (Group II), and 1.5 ml/g/min (Group III). Blood gases, hematocrit, and glucose were measured over 180 min. Nucleotides, serum osmolarity, and electrolytes were also measured. Morphological changes were recorded at the end of each experiment. Results. Acid base balance was better in livers from Group III. Electrolytes, osmolarity, hematocrit, and glucose showed no differences. Higher energy charges correlated with higher blood flow rates. On histopathology, there was evidence of more hydrostatic pressure-related changes in Group III, with no difference in necrosis or inflammatory-related alterations. Conclusions. (1) Energy charge correlates strongly with blood flow rate. (2) A rate of 1.5 ml/g/min allows for a better electrolyte, osmolarity, and acid-base balance. (3) Morphological analysis is not a sensitive predictor of organ viability as assessed by energetic potential. (4) The minimal blood flow rate required to attain an energy charge above 0.75 is 1.33 ml/g/min. PMID- 11421603 TI - Glutamine enhances the gut-trophic effect of growth hormone in rat after massive small bowel resection. AB - Background. Bowel-specific nutrient, glutamine, growth hormone, and modified diet have been reported to jointly improve nutrient absorption in patients with short bowel syndrome. However, controversy exists about the exact treatment factor. In this study we attempted to analyze the individual and combined effect of glutamine supplementation and growth hormone on small bowel adaptation by using tube feeding to control luminal nutrition supply. Materials and methods. Thirty two adult male Sprague-Dawley rats (278 +/- 8 g) underwent 85% mid-small bowel resection and were randomly assigned to four groups: Control, receiving control liquid diet via a gastrotomy tube; GLN, receiving liquid diet enriched with 20 g. L(-1) glutamine; GH, receiving subcutaneous growth hormone (GH) (0.3 IU, bid); and GLN + GH, receiving both glutamine supplementation and GH treatment. All animals were provided with isocaloric (60 kcal/day) and isonitrogenous (0.686 g/day) nutrition. Absorption tests were performed in the form of oral nutrient tests with (14)C-labeled glucose and (3)H-labeled palmitic acid on the Postoperative Day 12. Results. GH treatment significantly increased the plasma insulin-like growth factor I (IGF-I) level, body weight, jejunal and ileal villous height and mucosal thickness, and peak plasma (14)C and (3)H levels. Glutamine supplementation did not produce a significant difference; however, combined with GH treatment, glutamine supplementation further increased body weight, plasma IGF-I level, jejunal and ileal villous height and mucosal thickness, and peak plasma (14)C and (3)H levels significantly. Conclusions. After massive small bowel resection, enteral glutamine supplementation alone has no remarkable beneficial effect on bowel adaptation. However, glutamine supplementation enhanced the gut-trophic effect of GH. PMID- 11421604 TI - Nonlinear analysis of arterial oscillated flow in experimental stenosis and microsurgical anastomosis. AB - BACKGROUND: Thrombotic vascular occlusion is one of the main complications that can occur during microsurgical anastomosis and is frequent when the blood becomes turbulent. The aim of this ex vivo study was to test the use of nonlinear mathematical tools to detect turbulence flow upstream and downstream of an arterial stenosis and of a microsurgical anastomosis technique in arteries with diameters in the range of microsurgical practice. MATERIALS AND METHODS: Rat carotid arteries (0.8 to 1.2 mm diameter) were transferred to a flow chamber and perfused with Krebs solution. An oscillated vascular flow was initiated with a peristaltic pump and a transit time flowmeter was used to measure flow with two probes. An arterial stenosis was created by a ligature and progressively increased ranging from 0 to 95%. For each flow signal three nonlinear analytical procedures were applied: time-delayed procedures, correlation dimension, and computing of the largest Lyapunov exponent. RESULTS: Our results indicated that the level of turbulence flow is correlated with the area reduction stenosis. In the range of 60-95% area reduction stenosis, we noted an experimental increase of turbulence flow. We also founded that a classical end-to-end anastomosis technique induced an increase of the turbulence flow in comparison with a control artery. CONCLUSIONS: Thus nonlinear analysis can be useful in characterizing the complexity of an oscillated flow in small arteries submitted to stenosis or microsurgical anastomosis and may have clinical uses in detecting high level turbulent flow after microsurgery. PMID- 11421605 TI - Blocking CTL-based cytotoxic pathways reduces apoptosis of transplanted hepatocytes. AB - BACKGROUND: A major obstacle in allogenic hepatocyte transplantation is increased apoptosis of grafted cells due to CTL-based cytotoxicity. However, whether blockade of Fas- and granzyme-mediated pathways of CTL-based cytotoxicity may provide immune protection to transplanted hepatocytes is poorly defined. Our study aimed to reduce apoptosis of allogenic transplanted hepatocytes by inhibiting granzyme B (GraB) activity and blocking Fas-FasL interaction. MATERIALS AND METHODS: Hepatocyte transplantation was performed by inoculating isolated liver cells from ACI rats (allogenic) or Lewis rats (syngenic) into the spleens of Lewis rats. Recipients were treated with FLIM58, an inhibitory anti FasL mAb, and GraB inhibitor I alone or a combination of the two drugs for 5 days after transplantation, and were sacrificed at Day 7. Apoptosis of transplanted hepatocytes was detected in situ by TUNEL assay and M30 immunostaining. Glutamate dehydrogenase (GLDH) activity in recipient spleens was examined to evaluate survival of transplanted cells. Recipient spleens were assayed for FasL level with Western blotting and for GraB activity by hydrolysis of GraB substrate. RESULTS: FLIM58 or GraB inhibitor I significantly reduced the percentage of TUNEL positive and M30-positive hepatocytes and markedly increased GLDH levels in allogenic, but not syngenic, recipient spleens. These effects were more pronounced when the two drugs were used in combination (P < 0.05). Additionally, elevation of FasL and GraB levels in allogenic recipient spleens can be significantly reduced by FLIM58 and GraB inhibitor I, respectively. CONCLUSIONS: Inhibition of GraB activity and blockade of Fas-FasL interaction reduce the apoptosis of allogenic transplanted hepatocytes, and thus improve their survival. PMID- 11421606 TI - Affinity of Staphylococcus epidermidis to various prosthetic graft materials. AB - BACKGROUND: Abdominal wall hernias have always been a major problem for general surgeons. The techniques of repairing primary, recurrent, and incisional hernias have evolved throughout the years at an accelerating trend, especially after production of prosthetic graft materials. Although looked upon with suspicion due to infection, fistula formation, and foreign body reaction, prosthetic graft materials are used deliberately in primary and recurrent hernias. The present study was designed to evaluate bacterial adherence to frequently used prosthetic graft materials. MATERIALS AND METHODS: The study was carried out in five different groups with each group consisting of 10 identical samples of the same kind of prosthetic graft material. The prosthetic graft materials used in the study were polypropylene, polyglactin 910, polyester fibers, steel, and polytetrafluoroethylene (PTFE). These prosthetic graft materials were incubated in vitro with a Staphylococcus epidermidis strain which was ++++ adhesion positive. The degree of adhesion of S. epidermidis to prosthetic graft materials was assessed by the ELISA method. RESULTS: Vicryl grafts showed significantly minimal bacterial adhesion whereas PTFE grafts tended to have more adhesion but this did not reach a statistical significance. Other graft materials did not show any difference for bacterial adhesion (Table 3). CONCLUSION: These results suggest that in vitro S. epidermidis adhesion to Vicryl grafts is less than other types of prosthetic graft materials (P < 0.05 for all comparisons). Further in vitro and in vivo studies are required to confirm these results and to understand the complex interactions among bacteria, graft material, microenvironment, and surgical technique. PMID- 11421607 TI - In situ detection of hTERT mRNA relates to Ki-67 labeling index in papillary thyroid carcinoma. AB - BACKGROUND: Telomerase is activated in most human cancers but is inactivate in adult somatic tissues except for some proliferating cell lineages. The maintenance of telomerase activity may be a critical step of cellular immortalization and transformation. MATERIALS AND METHODS: We analyzed the expression of human telomerase reverse transcriptase (hTERT) using in situ hybridization and compared it to Ki-67 immunoreactivity in 29 cases of papillary thyroid carcinoma (PTC) and 17 cases of benign thyroid disease. RESULTS: The hTERT messenger RNA (mRNA) was expressed in the cytoplasm of carcinoma cells with moderate (n = 10) to strong intensity (n = 10) in 69% (20 of 29) PTC cases. Human TERT was found in only 29% (5 of 17) cases of benign thyroid disease. Human TERT gene expression was preferentially detected in PTC (P = 0.021). The Ki-67 labeling index was observed in 16 cases of PTC (16 of 29; 55.2%). This result was significantly different from that of benign thyroid disease (P = 0.014). The Ki 67 labeling index related to the intensity of hTERT mRNA expression (r = 0.51; P = 0.005) and was inversely associated with the follicular variant of PTC (r = 0.413; P = 0.026). No statistically significant difference was found between hTERT expression and histological subtype of PTC. CONCLUSIONS: Our results demonstrated that expression of hTERT could be detected using in situ hybridization in PTCs and was significantly distinguishable from that of benign thyroid disease. Human TERT expression was related to the Ki-67 labeling index, indicating that coupling of telomerase activation with cell proliferation was the associated mechanism for tumorigenesis. PMID- 11421608 TI - Interleukin-8 administration enhances venous thrombosis resolution in a rat model. AB - BACKGROUND: Therapy for deep vein thrombosis (DVT) resolution in those patients in whom a complication or contraindication to anticoagulation occurs is limited. As prior work suggests that thrombus maturation involves early influx of neutrophils (PMN) and neovascularization, we hypothesized that administering the proinflammatory/proangiogenic chemokine interleukin (IL)-8 might accelerate thrombus resolution. MATERIALS AND METHODS: An established rodent model of DVT (inferior vena cava [IVC] ligation) was used whereby daily intravenous recombinant human IL-8 (1 microg) or vehicle control was administered, with sacrifice at 4 and 8 days. Prior to sacrifice and at harvest, duplex ultrasound of the DVT and femoral venous pressure measurements were performed. Thrombi were analyzed by immunohistochemical techniques for PMN, monocytes, and neovascularization; for chemokines, by enzyme-linked immunoassay; and fibrosis, by hydroxyproline assay and trichrome staining. RESULTS: IL-8 accelerated thrombus dissolution 4 days after IVC ligation, with 6-fold increased thrombus blood flow by duplex ultrasound and a 23% increased absolute femoral venous pressure compared with controls (both P < 0.05). These findings may be partially explained by the fact that animals receiving IL-8, as compared with controls, had 2.5-fold greater thrombus neovascularization (with a trend continuing to 8 days) and increased PMN at 4 days. Thrombus vascular endothelial growth factor was significantly reduced at 8 days postligation, while monocyte chemotactic protein 1 and macrophage inflammatory protein-1alpha were not altered by IL-8 administration. At 8 days post-IVC-ligation, fibrosis was 12-fold greater with IL 8 treatment compared with controls. CONCLUSIONS: A proinflammatory/proangiogenic thrombus milieu, as conferred by IL-8, enhances thrombus resolution and underscores the important relationship between neovascularity and inflammation. PMID- 11421609 TI - The initiating factors of late preconditioning in skeletal muscle. AB - PURPOSE: The goal of these studies was to determine the initiating factors for late preconditioning in the microcirculation of skeletal muscle. MATERIALS AND METHODS: The cremaster muscle of male Sprague-Dawley rats underwent 4 h of ischemia and then 60 min of reperfusion. Ischemic preconditioning (IPC) consisted of 45 min of ischemia but was done 24 h before the 4 h of ischemia. To mimic the effects of IPC in the late phase, adenosine (ADO) or sodium nitroprusside (SNP) was given 24 h before the prolonged ischemia via local intraarterial infusion. To block the effects of IPC in the late phase, 8-sulfophenyl-theophylline (a nonspecific ADO receptor blocker) or N(W)-nitro-l-arginine (a nonselective nitric oxide synthase antagonist) was given prior to IPC. Microvascular response to IPC and pharmacological preconditioning were determined by measuring arteriole diameters and capillary perfusion using intravital microscopy. RESULTS: Administration of ADO or SNP on day 1 without IPC produced a similar microvascular protection against prolonged ischemia/reperfusion on day 2 as that induced by IPC alone. In contrast, blocking ADO receptors or nitric oxide synthase on day 1 just prior to IPC eliminated the IPC-induced microvascular protection seen on day 2. In addition, inhibition of nitric oxide synthase on day 1 diminished the protection induced by ADO, but blocking ADO receptors on day 1 did not compromise the protection induced by SNP. CONCLUSION: The results from these studies suggest that up regulation of ADO is the initiating factor with secondary up regulation of nitric oxide in late preconditioning. Both ADO and nitric oxide contribute to initiating microvascular protection in the late phase of IPC. PMID- 11421610 TI - Inhibition of neointimal formation in porcine coronary artery by a Ras mutant. AB - BACKGROUND: Therapeutic approaches to reduce the neointimal formation caused by balloon injury have been focused mainly on experimental models of restenosis in the rat carotid artery. However, restenosis in rat carotid artery may not replicate the coronary arterial responses to injury in larger animals and humans. METHODS: In this study, we used pig coronary arteries as an animal model to evaluate the preventive effects of a virus-mediated dominant negative mutant RasN17 on balloon injury-induced restenosis. The viral particles were delivered to the balloon-injured coronary arteries via a dispatch catheter to keep the virus in a confined arterial segment for 10 min to reach optimal transfection. Six weeks after balloon injury, the pigs were sacrificed and the left anterior descending arteries were isolated for histological analysis. RESULTS: Neointima formation was prominent in the group receiving balloon injury as compared with the uninjured controls. A remodeling process with migration of collagen was also found in the injured coronary arteries. The application of AdRasN17 led to a 56% decrease in neointima formation and a 75% increase in lumen size, as compared with the balloon-injured vessels treated with AdLacZ control. CONCLUSIONS: These results suggest that AdRasN17 is an effective therapeutic gene in preventing balloon injury-induced neointimal formation in pig coronary arteries. PMID- 11421611 TI - Effects of profound hemodilution on small-intestinal wound healing in rabbits. AB - BACKGROUND: Wound healing is influenced by tissue oxygen tension and blood perfusion, but not by moderate anemia or hemodilution. The effect of perioperative profound hemodilution on small-intestinal wound healing remains unclear. METHODS: We performed jejunectomy followed by end-to-end anastomosis in rabbits subjected to a variety of perioperative hemodilutions: HD((HES)), hemodiluted with hydroxyethylstarch; HD((P+HES)), hemodiluted with autologous plasma and hydroxyethylstarch; HD((HES))/R, hemodiluted with hydroxyethylstarch and retransfused afterward. Intraoperative hemoglobin levels were 5 g 100 ml(-1). On Postoperative Day 5, the tensile strength (TS) of the anastomosis was measured and histological specimen was obtained. The time courses of hemoglobin, serum albumin (Alb), plasma fibrinogen (Fbg), and plasma activity of factor XIII (F XIII) were measured. RESULTS: TS in HD((HES))/R (236.0 +/- 52.2 gf) was similar to that in control (266.5 +/- 41.6 gf); however, TS in HD((HES)) (179.8 +/- 17.9 gf) and HD((P+HES)) (165.5 +/- 14.7 gf) decreased significantly. The histological findings in HD((HES))/R were similar to those of control, whereas they demonstrated a delayed healing process in HD((HES)) and HD((P+HES)). Hemoglobin levels were still lower on Postoperative Day 5 in HD((HES)) and HD((P+HES)), but increased to 10.0 g 100 ml(-1) after retransfusion in HD((HES))/R. Hemodilution caused significant decreases in Alb, Fbg, and F XIII, but the values after retransfusion in HD((HES))/R were similar to postoperative values in HD((P+HES)). CONCLUSION: Intraoperative profound hemodilution does not interfere with small intestinal wound healing as long as postoperative hemoglobin levels were maintained above 10 g 100 ml(-1). Postoperative levels of other plasma constituents may not influence wound healing. PMID- 11421612 TI - Intestinal and hemodynamic impairment following mesenteric ischemia/reperfusion. AB - BACKGROUND: Clinical intestinal ischemia/reperfusion (I/R) injury results in local and systemic dysfunction. A rat model of transient mesenteric occlusion has been used to study this phenomenon. However, a systematic analysis of the rat model with respect to intestinal permeability and hemodynamics has not been carried out. MATERIALS AND METHODS: In anesthetized rats, the superior mesenteric artery was occluded for 60 min, followed by reperfusion for 4 h. Intestinal impairment was evaluated via histological examination and by measuring ex vivo apparent permeability coefficients (Papp) of mannitol (0.18 kDa), inulin (5 kDa), and dextran (70 kDa). Hemodynamic effects of intestinal I/R were determined by monitoring mean arterial pressure (MAP) and heart rate (HR) via a catheter placed in the femoral artery. RESULTS: The animal model was associated with increased ex vivo Papp for mannitol and inulin. Although I/R injury was accompanied by significant histological disruption, there was no observable alteration in dextran permeability, suggesting that the loss in normal barrier function was limited to low-molecular-weight compounds. Hemodynamic measurements indicated that reperfusion induced a precipitous and sustained fall in MAP. HR values fell sharply following reperfusion but gradually increased and eventually "overshot" to values greater than baseline. CONCLUSIONS: Our findings demonstrate the selective loss of barrier function of the small bowel following intestinal I/R. Furthermore, these results also illustrate the importance of selecting appropriate permeability markers for the evaluation of intestinal damage. In light of the significant hemodynamic disruption accompanying the animal model, our investigation also points toward the need for developing therapeutic strategies that mitigate the local and systemic effects of intestinal I/R injury. PMID- 11421613 TI - Intestinal mitochondrial dysfunction in surgical stress. AB - BACKGROUND: Surgical stress is associated with altered intestinal function. Our earlier study using a rat model indicated that oxidative stress plays an important role in this process. Since mitochondria are crucial to cellular function and survival and are both a target as well as a source of reactive oxygen species, the present study looks at the changes in enterocyte mitochondria during surgical stress. METHODS: Surgical stress was induced by opening the abdominal wall and handling the intestine as done during laparotomy. Mitochondria were prepared from the isolated enterocytes at different time periods after surgical stress. The effect of surgical stress on enterocyte mitochondrial ultrastructure, respiration, anti-oxidant enzyme activity, thiol redox status, calcium flux, permeability, and matrix enzymes was then studied. RESULTS: Surgical stress resulted in alterations in mitochondrial respiration and thiol redox status. It was also associated with altered mitochondrial matrix enzyme activity, decreased superoxide dismutase activity, induction of mitochondrial permeability transition, and swelling, as well as impairment of mitochondrial calcium flux. These alterations were seen at a maximum of 60 min following surgical stress and were reversed by 24 h. CONCLUSIONS: Laparotomy and mild intestinal handling itself results in enterocyte mitochondrial damage. Since mitochondria are important cellular organelles, this damage can probably lead to compromised intestinal function. PMID- 11421614 TI - Heat shock protein 27 inhibits apoptosis in human neutrophils. AB - BACKGROUND: Prolonged neutrophil(PMN) survival has been implicated in tissue injury following sepsis. A variety of bacterial products have been identified which inhibit PMN apoptosis including lipopolysaccharide(LPS). Extracellular heat shock proteins(Hsp) have recently been identified as potent regulatory signals for the innate immune system during the inflammatory response. We hypothesized that Hsp 27 can affect PMN phenotype with respect to apoptosis and cytokine profile. MATERIALS AND METHODS: PMN were isolated from the peripheral blood of healthy human volunteers by red blood cell sedimentation and gradient centrifugation. Cells were placed in media and cultured for 18 h with and without recombinant human Hsp 27 at various concentrations. In parallel experiments, PMN were stimulated with LPS, a known inhibitor of PMN apoptosis, for comparison. Apoptosis was quantified using annexin V and propidium iodide staining with flow cytometric analysis. Culture supernatants were assayed for secretion of TNF alpha, IL-10, and IL-12. RESULTS: Hsp 27 significantly inhibits PMN apoptosis [control; 81.8 +/- 3.6%, vs Hsp 27, 60.4 +/- 4.1% p < 0.05]. The reduction is similar to that signaled by LPS, alone. Together their effect is not synergistic. The Hsp 27 response is dose-dependent. Hsp 27 does not induce secretion of TNF alpha, IL-10, or IL-12, whereas LPS does signal IL-12 and TNF-alpha secretion. CONCLUSION: These data demonstrate that exogenous Hsp 27 may play a role in neutrophil-mediated tissue injury during trauma and sepsis via its ability to inhibit neutrophil apoptosis. However, Hsp 27 does not significantly alter neutrophil phenotype with respect to cytokine production profile. PMID- 11421615 TI - The role of tumor necrosis factor-alpha and interleukin-1beta in ischemia reperfusion injury of the rat small intestine. AB - BACKGROUND: To determine the role of tumor necrosis factor (TNF) and interleukin (IL)-1 in small-intestinal ischemia-reperfusion (I-R) injury, we investigated the effect of FR 167653, a specific IL-1 and TNF inhibitor, on warm I-R injury of the rat small intestine. MATERIALS AND METHODS: Male rats treated with either saline (NS group) or FR 167653 (FR group) underwent 150 min of warm small-intestinal ischemia by applying a vascular clip at the origin of the superior mesenteric artery. In addition to the survival analyses, we investigated plasma TNF-alpha and endotoxin levels, intestinal tissue TNF-alpha and IL-1beta levels, hematocrit values and the amount of exudates in the intestinal lumen, glutamic aspartate aminotransferase (AST), and histological findings up to 120 min after reperfusion. RESULTS: TNF-alpha and IL-1beta levels in the intestinal tissue, and plasma TNF-alpha and endotoxin levels, were significantly (P < 0.05) reduced in the FR group. Severe mucosal damage on histological findings (120 min after reperfusion) and a large amount of intraluminal exudates (60 min after reperfusion) were shown in the NS group, but these findings were significantly (P < 0.05) ameliorated in the FR group. Serum AST levels in the NS group increased 120 min after reperfusion, but this change was significantly (P<0.05) reduced in the FR group. The 30-day survival rate was 80% in the FR group and 30% in the NS group (P<0.05). CONCLUSIONS: Dual inhibition of TNF and IL-1 effectively alleviated intestinal I-R injury, suggesting the key role of TNF and IL-1 in this pathophysiology. PMID- 11421616 TI - Fetal therapy with rhIGF-1 in a rabbit model of intrauterine growth retardation. AB - BACKGROUND: Intrauterine growth retardation (IUGR) may, in part, be due to a deficiency of insulin-like growth factor-1 (IGF-1). The objectives of this study were to determine the relationship between fetal serum IGF-1 levels and fetal and placental size in a rabbit model of IUGR and to compare two techniques of selective, exogenous IGF-1 administration (transamniotic and branch uterine arterial catheter infusion) to growth-retarded fetuses in utero. MATERIALS AND METHODS: Pregnant rabbits (n = 6) had their fetuses harvested near term (31 days) for fetal and placental weighing and serum collection. Growth-retarded fetuses were selectively infused for 7 days with recombinant human IGF-1 (rhIGF-1; 1,440 ng/day) either through a transamniotic catheter (n = 8) or via an adjacent uterine arterial branch catheter (n = 6). Opposite horn runts were sham catheterized, but not infused. At term, the fetal runt pairs and their placentas were harvested and weighed, and their serum was collected. The correlation between fetal and placental weight and endogenous serum IGF-1 was calculated (Pearson coefficient, r), while paired t-tests were used to compare the means between the IGF-1-infused and control groups. RESULTS: There was a significant correlation between fetal (r = 0.4230; P = 0.022) and placental weight (r = 0.4166; P = 0.025) and endogenous serum levels of IGF-1. Transamniotic infusion of rhIGF-1 was associated with an increase in serum IGF-1 level (254 +/- 79 vs 351 +/- 101 ng/ml, P = 0.04) and placental weight (5.4 +/- 2.3 vs 7.1 +/- 3.2 g, P = 0.005), and with a trend toward increased fetal weight between matched fetal runt pairs. Fetal mortality in the uterine arterial catheterized group was 76%, and there was no significant difference in fetal or placental weight or IGF-1 levels between infused and noninfused survivors. CONCLUSIONS: Endogenous fetal serum levels correlate with fetal and placental size in the rabbit IUGR model. Transamniotic administration of rhIGF-1 significantly increases serum IGF-1 levels and placental weight of fetal runts, while uterine vessel catheterization results in prohibitive fetal mortality and does not increase fetal or placental growth or IGF-1 levels. PMID- 11421617 TI - Hypoxia regulates osteoblast gene expression. AB - Vascular disruption secondary to fracture creates a hypoxic gradient of injury wherein the oxygen tension at the center of the wound is very low. In vivo this hypoxic microenvironment stimulates the expression of a variety of cytokines from inflammatory cells, fibroblasts, endothelial cells, and osteoblasts. In order to begin to dissect this complex system, we have examined the effects of hypoxia on isolated osteoblast gene expression in vitro. Understanding gene expression in this system may facilitate the development of targeted therapeutic modalities designed to accelerate fracture repair and reduce complications. Using an established model of in vitro hypoxia, we have analyzed the expression of genes involved in bone matrix production and turnover. Subconfluent neonatal rat calvarial osteoblasts were exposed to hypoxia (pO(2) = 35-40 mm Hg) and total cellular RNA was collected at 0, 3, 6, 24, and 48 h. Northern analysis was used to analyze the expression patterns of (1) transforming growth factors (TGFs) beta1, -beta2, and -beta3 and their type I receptor; (2) collagens I and III; and (3) tissue inhibitor of metalloproteinase-1. We have demonstrated a marked elevation of TGF-beta1 gene expression within 3 h of hypoxia. Although neither TGF-beta2 nor TGF-beta3 expression was affected by hypoxia, the TGF-beta type I receptor was substantially upregulated within 6 h. In addition, extracellular matrix scaffolding molecules (collagens I and III) were markedly, but differentially, upregulated. Finally, we have demonstrated that the expression of an inhibitor of extracellular matrix turnover, the tissue inhibitor of metalloproteinase-1, was strikingly decreased in response to hypoxia. These results imply that hypoxia can affect osseous healing by altering the expression of cytokines, bone-specific extracellular matrix molecules, and their regulators. PMID- 11421618 TI - Range of motion physiotherapy reduces the force deficit in antagonists to denervated rat muscles. AB - BACKGROUND: We used a rat hindlimb model of tibial nerve transection to determine if a loss of mechanical function exists in innervated antagonists compared with denervated muscles. We tested two hypotheses: (1) denervation of the rat ankle plantar flexors results in decreased force production of the ankle dorsiflexors, and (2) daily passive ankle range of motion (ROM) physiotherapy prevents or reduces the force deficit. METHODS: Adult Lewis rats were assigned to one of three groups: (1) a sham (S) group, in which the tibial nerve was exposed but not transected; (2) a no rehabilitation (NR) group, in which a 2-cm segment of tibial nerve was excised at midthigh to denervate the ankle plantar flexors; or (3) a rehabilitation (R) group, in which a 2-cm segment of tibial nerve was excised and the animals were subjected to ankle passive ROM physiotherapy for two 5-min sessions each day. After 14 days, maximum isometric tetanic force (F(0)) and specific force (sF(0)) were measured in the extensor digitorum longus (EDL) muscle, an ankle dorsiflexor. RESULTS: Compared with those from animals in the S group, EDL muscles from animals in the NR group demonstrated a 22% decrease in both F(0) and sF(0). In the EDL from animals in the R group, daily passive ROM physiotherapy diminished the deficit in F(0) but not in sF(0). CONCLUSIONS: These data support the hypotheses that nerve injuries result in impaired mechanical function in the innervated antagonists to denervated muscles and that passive ROM physiotherapy can improve force production in these muscles. PMID- 11421619 TI - Sudden infant death syndrome Part 2: the response of the reticuloendothelial system to hypoxemia and infection. AB - The incidence of ascites in chicks raised in a high-altitude chamber doubled from 6500 feet to 8000 feet. A similar condition developed in calves transported to pasture at high altitude. Chicks raised in a high-altitude chamber (compared to controls) produced more plasma cells in the germinal centres of the spleen about four days after an antigen challenge. Children usually suffering from a mild respiratory infection at sea level often developed pulmonary edema (HAPE) on transfer to high altitude. Sudden infant death syndrome (SIDS) victims produced more plasma cells in the germinal centers of the spleen. In one survey of SIDS, about half of the infants suffered an upper respiratory tract infection in the two weeks prior to death and the lungs were filled with fluid at autopsy. Elevated levels of hypoxanthine indicated hypoxemia before death, and a presumed response to hypoxemia in SIDS was the presence of extramedullary hematopoiesis in the liver. The effect of prolonged hypoxemia and infection are additive in increasing vascular permeability and the accumulation of edema fluid. The preferential uptake of zinc by edema fluid proteins at the expense of inflammatory cells increases the motility and metabolism of zinc-deprived activated macrophages. Activated macrophages release cytokines which in turn stimulate the release of pro-inflammatory peptides which increase vascular permeability and mortality. These inflammatory peptides are under proteolytic control. The neutral endopeptidase (NEP) is a cell-surface zinc metalloproteinase which modulates toxic shock.Zinc also modulates the inflammatory response of the activated macrophage. Interleukin-12 (IL-12), predominantly a product of macrophages, is involved in regulating both hematopoiesis and the adaptive immune response. IL-12 promotes interferon gamma (IFNgamma) production by T cells. IFNgamma acts on macrophages to release large amounts of nitric oxide (NO). An elevated immune response leads to NO overload, dilation of the cardiovascular system and toxic shock. A mechanism resulting in cardiovascular failure and a shock-like sequence is described in some cases of SIDS.Bradycardia, recorded on cardiorespiratory monitors in six SIDS infants, was considered a late event. Cytokines regulate all aspects of the immune response. Extramedullary hematopoiesis in the liver was one anatomical marker of hypoxemia in SIDS. This survey traces the function of the activated macrophage with the cytokines regulating extramedullary hematopoiesis and the precocious immune response in SIDS. PMID- 11421620 TI - Attempts to identify a control system for chemical reactivity in the living state using virtual energy. AB - This thesis explores the activation of chemicals in metabolic systems from the viewpoint that this activation is under the control of elements of the space-sea in which the chemicals are immersed. Themselves inert, the chemicals are theorised to exploit a force or action issuing from space (fluctuation) and characterized by the homogeneity (termed symmetry) of this medium. The fluctuation is heterogenized upon collision with matter from the intervention of well recognized fields of gravity and electromagnetism at the instant of its issue to form the near field of radiation. Fractions of original space waves and of their intrinsic spin are produced resulting in the activation of the orbitals (valency) in the chemical itself. The thesis continues: the disturbed fluctuation must return to space, obliging in turn, a prior return to the homogeneous state requiring special restorative wave rearrangements known as resonance. The success of the restorative resonance is signalled by a singularity of the fluctuation now propelled to infinity (space), and the contingent chemical reactions thereby terminated. Compromise to this return can occur from many causes and, in its presence, activation of the orbitals continues. They now effectively constitute autonomous reactions alienated from the system as a whole. The thesis is supported from evidence from diverse fields such as space theory, history of quantum field theory in attempts to derive its meaning, dielectrics and the near field of electromagnetic radiation, electron-space interactions at the Fermi surface during phase transitions and evolution of equilibrium conditions in resonance phenomena. The utility of the hypothesis rests on recognition of the resonance condition at various points in the system sufficiently macroscopic as to be available clinically as an abrupt interface between physiology and pathology. PMID- 11421621 TI - A control system for chemical reactivity in the living state: supportive evidence from laboratory studies. AB - This paper supports a thesis on control of chemical systems occurring in metabolism which makes extensive use of energy forms with an indirect or vicarious route to their targets in the chemicals concerned. In the recent decade or so, quantum chemists in studying the same problem, control of chemical reactions by indirect routes, not necessarily with a specific bio-function goal, have produced almost identical protocols known as quantum sculpting of the chemical wave function in their laboratory experiments. Parallels and modulations of the two approaches, native and man-made, are discussed in this paper with the aim of supporting the hypothesis at a level displaying high rationale for an intransigent aspect of physical chemistry of eventual clinical implication, that of control of complex chemical systems. PMID- 11421622 TI - Does an ultra violet photooxidation of the manganese-loaded/copper-depleted prion protein in the retina initiate the pathogenesis of TSE? AB - Ecosystems supporting clusters of sporadic transmissible spongiform encephalopathy (TSE) are characterized by common properties of high-manganese/low copper, zinc, selenium mineral status, and high-altitude/snow-covered/pre cambrian mountain terrain where above-average intensities of ultra violet/ozone oxidants are prevalent. Cell culture trials have confirmed the hypothesis that manganese (Mn) substitutes at Prion Protein's (PrP's) vacated copper (Cu) domain, whereupon PrP loses its Cu-mediated antioxidant function, transforming into a protease-resistant misfolded isoform that aggregates into fibril 'tombstone' structures - the key hallmark distinguishing TSE central nervous system (CNS) pathology. The cellular localisation of PrP suggests PrP serves a 'front line' contributory role in neutralizing radicals generated by incoming environmental oxidants, whilst an intensive expression of PrP messenger ribonucleic acid (mRNA) in the retina, melanocytes, epidermis, etc., suggests PrP performs a key antioxidant role as a 'photooxidative shock absorber'; binding of porphyrin IX, Congo red and other photosensitisers to PrPc suggests PrPc serves as an integral associate of the porphyrin/melanin chromophore electron transfer chain; thereby serving as a quencher of singlet O2/superoxide generated by photoenergised chromophores/xeno photosensitisers. It is proposed that sporadic TSE pathogenesis is initiated in the retina of environmentally/genetically predisposed individuals via a two-stage chronic toxic process - Mn substitution at PrP's Cu domain forming a stable Mn2+-PrP complex, followed by an ultra violet in situ photo oxidization of the Mn2+ component; whereby the latent 'Jekyll and Hyde' capacity of the Mn2+-PrP conjugate is activated into the fully fledged, 'infectious' lethal auto-oxidizing, Mn3+-PrP 'prion' agent. Thus, PrPc's Cu-mediated antioxidant function is replaced by a Mn3+-mediated autooxidant dysfunction. Could the UK's increased loading of a cocktail of environmental oxidants that penetrated the CNS of the UK bovine (ultra violet microwaves/ozone/systemic cu chelating insecticides) account for a more virulent Mn4+ mediated acceleration of the TSE degenerative process in Mn-contaminated/genetically predisposed individuals, manifesting as the widespread emergence of new-variant bovine spongiform encephalopathy (BSE)/variant Creutzfeldt-Jacob disease (VCJD)/FSE in younger mammals? PMID- 11421623 TI - Sickle cell disease: a chronic inflammatory condition. AB - Homozygous sickle cell disease (SCD) has a wide spectrum of clinical manifestations which varies from an almost asymptomatic condition to severe illness, despite the fact that all subjects with this disease have the same base change in their DNA. The source of this variation is partly environmental, but a large part of this variability can derive from the presence of genetic modulators which are not fully understood. It was postulated that some degree of immunodeficiency should be associated with this condition, but no deficiency, directly related to a given component of the immune system, was observed that could explain the high levels of recurrent infections presented by sickle cell disease patients. Reviewing data from the literature we suggest that the influence of the immune system in the variation of clinical manifestations presented by SCD patients is not related with any immunodeficiency but is rather the result of a chronic inflammatory condition. PMID- 11421624 TI - Hunting genes in Parkinson's disease from the roots. AB - Parkinson's disease (PD), a common, neurodegenerative disorder, has a worldwide distribution. The genetic basis of PD is not well understood, although some recent leads have emerged. Epidemiological studies suggest that there is significant variation in the prevalence of PD between different populations and rates are highest in populations of European origin. Significant differences in molecular pathology in PD and control brain tissue have been observed between African, British and Indian populations. In view of this epidemiological and pathological evidence, it is proposed that allelic variations in genes that predispose to PD may account for the ethnic variation. Advances in our knowledge about the human genome will allow us to make detailed comparisons between affected and control subjects in different populations. This may help us to understand the reasons for the variation, and a better understanding of the genetic processes underlying the disease process. PMID- 11421625 TI - Hydrogen peroxide: a potent cytotoxic agent effective in causing cellular damage and used in the possible treatment for certain tumours. AB - H2O2, a highly reactive agent, can react under certain conditions with a variety of cellular components. These reactions include the lipid peroxidation of membrane and hydroxylation of proteins and DNA. The reactions can take place in the presence of oxygen and are fairly rapid, the H2O2 being converted to water and oxygen. Experiments were carried out in vitro to assess the ability of this agent to destroy cancer cells without generating dangerous by-products. The direct administration of aqueous H2O2 into solid tumours has the potential to cause tumour cell death. The efficacy of the use of H2O2 for treating 'solid' cancers will necessitate its delivery to the tumour site, for example by direct special multiple injection of H2O2 into a detectable tumour mass. We anticipate that, if suggested mode of delivery can be obtained, H2O2 can act as an anti cancer drug with two distinct advantages over conventional chemotherapeutic agents: to produce minimal short- and long-term side-effects and is relatively cheap and cost effective. PMID- 11421626 TI - Iron loading: a risk factor for Whipple's disease? AB - Because of impairment of microbial iron acquisition ability, some potential pathogens can cause disease only in iron loaded hosts. Tropheryma whippelii, the etiologic agent of Whipple's disease, is a possible example. Whipple's disease is non-contagious, occurs mainly in middle-aged white males, and displays many, but not all, of the complications of hereditary haemochromatosis. Tropheryma whippelii is a gastrointestinal commensal that causes disease in persons who have a Th1-Th2 imbalance. Host susceptibility may be exacerbated by iron loading. Consideration should be given to have patients evaluated for levels of interferon gamma and interleukin-4 as well as for serum ferritin and transferrin iron saturation. PMID- 11421627 TI - Increased facial temperature as an early warning in Sudden Infant Death Syndrome. AB - The promotion of supine sleeping position in young infants has resulted in significant declines in the incidence of Sudden Infant Death Syndrome although little is understood in terms of mechanisms. We hypothesize that supine sleeping position promotes appropriate thermal regulation via the face and head which is the major source of infant heat loss. By facilitating temperature regulation, the supine position ensures that the centre for thermoregulation in the hypothalamus does not become dysfunctional due to local temperature fluctuations. Because these hypothalamic, thermoregulatory neurones are synaptically linked to those regulating respiration in the medulla, adequate temperature control by the infant maintains normal respiration. In contrast, an increase in face and head temperature over and above core temperature would suggest thermoregulatory stress and an increased likelihood of respiratory apnoea. PMID- 11421628 TI - Diabetes: energetics, development and human evolution. AB - The recent emergence of the thrifty phenotype as an explanation for metabolic efficiency has brought evolutionary perspectives on diabetes, as represented by the thrifty genotype, under scrutiny. However, the logic of natural selection along with evidence from non-human primates supports the role for energetic constraints in the evolution of metabolic efficiency, particularly in skeletal muscle physiology. Environmental fluctuation during human evolution would have provided selective pressures for the development of efficient skeletal muscle starting prenatally and continuing throughout the lifespan. Such mechanisms including, glucose transporters, mitochondrial gene expression, leptin receptors and uncoupling proteins, should be present in all humans, though some living populations may exhibit particular 'thriftier' alleles. A focus on physical activity and the factors underlying efficient muscle physiology has implications for prevention of diabetes in both developing and developed societies. PMID- 11421629 TI - Cognitive theory and illness behavior in disability syndromes. AB - There are many controversial disability syndromes, representing medicolegal and social dilemmas for a variety of medical disciplines. While illness behavior and sick role phenomena are often invoked to explain many of these syndromes, the extent to which such phenomena are under volitional control has not been thoroughly explored. The volitional control of illness behavior has important treatment implications, and may explain why cognitive therapy can be effective in these patients. Further understanding of the relevance of cognitive theory to illness behavior, the sick role, secondary gain, and disability may render even more effective cognitive therapy approaches. This review explores the consciousness states, the role of each state in information processing (in this case processing illness information), the automaticity and hence volitional state of each level of information processing, and the likelihood that illness behavior in disability syndromes is volitional. The cognitive model of these syndromes considers the interaction of automaticity, volition, and illness behavior and likely has numerous clinical, social, and legal applications. PMID- 11421630 TI - An insight into cortisol and polymyositis control with steroid therapy. AB - In the present report we have developed a mathematical model to describe the processes involved during high steroid therapy. We have shown that the model can predict clinical observations as well as determine the optimum steroid regimen without relying on trial and error methods. The model incorporates rate processes that simplify the physiological complexity as a few representative steps. In this way, it is possible to simulate clinical observations and then to predict clinical outcomes due to drug concentrations or additional reactions within the body. Specific examples are provided from patients with polymyositis and Becker's disease. For the first time, we provide an explanation into the control of cortisol production, and determine an optimum prednisolone concentration in order to minimize steroid side effects while maintaining adequate control of muscle degradation. PMID- 11421631 TI - Is induction of type 2 programmed death in cancer cells from solid tumors directly related to mitochondrial mass? AB - Many solid cancers respond to chemo or radiotherapy with a type 2 form of programmed cell death. This requires direct participation of mitochondria with release of cytochrome c and other factors that activate the 'execution' phase of the process. It is believed that as solid cancers progress, less differentiated clones containing fewer mitochondria evolve. Consequently, the mitochondrial 'switch' that activates the type 2 process will become less effective, as the number of elements available and their mass-effect declines. The opportunity for successful therapy, considered to depend upon the ability to activate programmed cell death, therefore becomes progressively less probable. PMID- 11421632 TI - A new application of rTMS: the sleeping brain and depression. AB - A new method of modifying brain function is rTMS (rapid transcranial magnetic stimulation) which has a positive effect on depression. Rapid transcranial magnetic stimulation during sleeping should be tested for a possible effect on sleep normalization. PMID- 11421633 TI - A bihormonal model of normal sexual stimulation; the etiology of premature ejaculation. AB - The physiological sexual excitation is mediated both by the sympathetic and the parasympathetic nervous system. The antagonism between testosterone (Tt) and estrogens (Es) as well as the fact that the dynamics of the sexual excitation in the male are antagonistic to those in the female are nowadays well-known; hence, the hypothesis was emitted that the sympathoparasympathetic sexual excitation is bihormonal mediated, consisting in a very active sexual hormone associated with a weak antagonistic hormone. Most studies show a serotonergic (ejaculatory) involvement in premature ejaculation (PE). Nevertheless, an effective treatment of PE is rather difficult with selective serotonin reuptake inhibitor drugs and other clinical data even suggest that changes in PE actually involve at first the sexual excitation process and only secondly the ejaculation reflex.Thus, a therapeutic model for PE is set up, starting from the physiological aspects described and from the presumed pathophysiological mechanism in PE. PMID- 11421634 TI - Apoptotic death by ceramide: will the real killer please stand up? AB - Many studies have shown that the addition of ceramide to an incubation medium, or procedures that lead to increased ceramide concentrations, can begin a process that leads to slowing of cell growth or apoptotic cell death. Only a few studies have examined the nature of the accumulating ceramide: is it composed of a fatty acid and sphinganine, or a fatty acid and sphingosine? Of the studies involving addition of ceramide to a cell culture, almost all have found that the sphingosine amide is active, not the sphinganine amide. Nearly all of these studies have utilized the rare form of ceramide, containing an acetyl group rather than the commonly found palmitoyl, stearoyl, or longer group. Acetyl sphingosine produces some unexpected effects with cells, the most striking being the formation of reactive oxygen species and mitochondrial damage. This mitochondrial damage appears to be an essential step in apoptosis. The possibility should be considered that the reactive oxygen species appear as the result of oxidation of the allylic alcohol group in unsaturated ceramides. This question is very relevant to a host of ceramide functions, particularly cancer chemotherapy and cell growth. PMID- 11421635 TI - Etiology and pathogenesis of anxiety disorders. AB - The author suggests that biological abnormalities related to the development of anxiety disorders can be classified as real or relative. Individuals with 'real abnormalities' are not able to function under any circumstances, including the circumstances that are natural for humans. Persons with 'relative abnormalities' can function under the natural circumstances, but are not able to function in situations that are unnatural for humans. Unnatural situations include being in elevators, flying airplanes, driving cars, etc. The author suggests that all humans can be classified into four groups: 1. Individuals with excellent 'adaptational reserve' never develop anxiety disorders; 2. Individuals with good adaptational reserve develop anxiety disorders only if they are subjected to extremely stressful, unusual situations (e.g. being a hostage); 3. Individuals with moderate adaptational reserve develop anxiety disorders if they are exposed to unnatural situations. They have 'relative abnormalities'; 4. Individuals with poor adaptational reserve develop anxiety disorders in situations that are natural for humans. They have 'real abnormalities'. PMID- 11421636 TI - DNA quadruplexes and dynamical genetics. AB - In a recent paper, we have put forward the hypothesis that there exist smart purposive mechanisms - tandem repeat length managers - which regulate the length of some tandem repeat, or cause rearrangements, and are almost always driven by some variable number tandem repeat. We have called the framework in which such mechanisms act 'dynamical genetics'. The purpose of this paper is to contribute to lay the foundations of a molecular study of the above mechanisms, by proposing a hypothesis, based on various kinds of supporting evidence and plausibility arguments, about the special importance of DNA quadruplexes for dynamical genetics, and by considering the involved enzymes. This hypothesis states that a tandem repeat length manager acts almost always by monitoring a DNA tract that has the characteristics of being a variable number tandem repeat and/or forming a DNA quadruplex, and that it is almost always driven by at least one of them. PMID- 11421637 TI - Cerebral malaria and developmental biology: making parasites grow-up. AB - Erythrocytes infected with the protozoan parasite Plasmodium falciparum, in the final stages of asexual development, sequester and adhere to the lining of capillaries in the brain; killing 1-3 million (mostly children under the age of 5) each year. I will argue here that the rapidly evolving tools of developmental biology be employed to find a way of inducing gametocytogenesis, thereby making the parasite 'grow-up' prematurely, and in large numbers alleviating the severe symptoms in the brain. PMID- 11421638 TI - Make PEEC, not war: does evasive transdifferentiation protect from carcinogenesis? AB - DNA repair and immune surveillance are established mechanisms that secure a programmed escape from the evolution of cancer (PEEC) by healing and elimination, respectively. Evasive transdifferentiation, i.e. silencing of carcinogenic events, is an emerging concept with potentially broad applicability that paradigmatically includes the claim that common acquired melanocytic nevi represent a programmed rescue from malignity for initiated melanocytes. This hypothesis is difficult to test. Research into the molecular mechanisms that govern the development of acquired melanocytic nevi, however, should result in the identification of a molecular master switch and an associated set of genes that - beyond the processes involved in apoptosis - may help bypass carcinogenesis and may be of therapeutic benefit in the future. PMID- 11421639 TI - Iron(III)-mediated intra-articular crystal deposition in arthritis: a therapeutic role for iron chelators. AB - Crystal deposition in arthritic diseases has attracted much interest. Many reports have established the presence of calcium pyrophosphate (CPPD), hydroxyapatite (HAP) and urate crystals throughout the range of arthritic diseases. In particular, HAP crystals have been detected in 30-60% of synovial fluid (SF) samples from patients suffering from osteoarthritis (OA) and 33% of those suffering from rheumatoid arthritis (RA). In OA, crystal deposition has been linked to greater joint deterioration. The mechanism of intra-articular calcification is unknown. Nucleation is required to transform a 'metastable' phosphate- and calcium-rich biofluid into one that generates crystals. Ferric ions have been demonstrated to induce crystallization of these stable supersaturated solutions via the process of nucleation. The inflamed arthritic joint is prone to iron loading. Microbleeding from compromised vasculature contributes to intra-articular iron loading in arthritic conditions. Low molecular-mass redox-active iron complexes have been detected in SF in inflammatory joint diseases. These species are credited with mediating oxidative stress via interaction with peroxides and superoxide. In addition, adventitious low-molecular-mass iron complexes can cause nucleation leading to crystal growth within the joint. Decorporating agents capable of removing this misplaced iron from the arthritic joint would have the joint benefit of relieving oxidative stress and preventing crystal nucleation. Systemic side effects could be overcome by the targeting suitable chelators using bioreductive delivery systems that are activated in hypoxic inflamed synovial tissue. PMID- 11421640 TI - Induction of immunological tolerance to the chemical carcinogens in early ontogenesis. AB - It is postulated that the induction of immunological tolerance to chemical carcinogens is due to the penetration of carcinogens and its albumin and DNA adducts from the maternal organism into the embryo. Maternal serum lgG-antibodies to carcinogens promote the transportation of carcinogens and adducts into the embryo through placenta and thus stimulate the tolerance induction. On the contrary, maternal secretory lgA-antibodies to carcinogens protect maternal, embryonal and new-born organisms from environmental carcinogens and thus inhibit the tolerance induction. The low individual maternal secretory lgA/serum lgG ratio leads to the immunological tolerance to carcinogens both in embryos and children. PMID- 11421641 TI - A pupillary aqueous pump. AB - Summary The existence of a pupillary aqueous pump moving the aqueous humour from the posterior to the anterior eye chamber is postulated. PMID- 11421642 TI - Electromechanical dissociation in the human skeletal muscle. AB - It is not clear whether electromechanical dissociation can be a significant cause of clinical weakness in the human skeletal muscle. One type of muscle disorder suggests that this may occur. The condition results from the use of large doses of corticosteroids in conjunction with a neuromuscular blocking agent. In this situation, the thick myosin filaments of the sarcomere are preferentially disrupted. This may selectively compromise the contractile properties of the skeletal muscle fiber, without necessarily implicating the integrity of the muscle membrane. Thus, a muscle fiber might still be able to depolarize electrically, producing a membrane action potential, without necessarily generating an appropriate mechanical response. PMID- 11421643 TI - Possible role of juxtaglomerular apparatus in low cardiac output syndrome and multiple organ failure: modulation by high sodium load. AB - Sympathetic overdrive in acute low cardiac output syndrome, diverts blood from cutaneous and visceral circulation centripetally. Microcirculation in general, and renal circulation in particular, deteriorates during these circulatory adjustments leading to multi-organ failure (MOF). Decreased afferent glomerular arteriolar blood flow, increased renal sympathetic nerve discharge and a resultant decreased sodium chloride delivery around macula densa stimulates the Juxta-glomerular apparatus (JGA) and triggers renin-angiotensin-aldosterone mechanism. This, along with increased ADH production results in a state of vasoconstriction, increased after-load, and continued fluid retention, further compromising the visceral microcirculation. Initially the fluid retention under the effect of aldosterone and ADH is iso-osmotic, but later under inappropriate ADH action more water than salt is retained, as evidenced by the presence of hyponatraemia and 'water-logging' in the endstage of this condition.The author hypothesizes that: although physiological, the persistent stimulation of the JGA during the low cardiac output state plays an important role in perpetuating a negative cardiovascular vicious cycle and further aggravating it into MOF. Furthermore, by infusing hypertonic saline and hence increasing the sodium chloride delivery to the distal tubules and the macula densa, the JGA could be inhibited. This strategy should work like angiotensin-converting-enzyme-inhibitor drugs in chronic cardiac failure, except by acting at the root cause and inhibiting Renin production at its source. It should further help by stimulating atrial natriuretic peptide secretion. PMID- 11421644 TI - Detection and taxonomic placement of endophytic fungi within frond tissues of Livistona chinensis based on rDNA sequences. AB - The 5.8S gene and flanking internal transcribed spacers (ITS1 and ITS2) of the rDNA were amplified from total DNA extracted from frond tissues of Livistona chinensis with universal and fungal-specific primers. These amplified fragments were cloned and sequenced. Phylogenetic analysis based on the 5.8S gene sequences indicated that the six clone sequences obtained were of different origins. Five sequences, P1-9, P2-6, P4-4, P4-5, and P4-7, belonged to the fungi and one sequence, P3-2, belonged to the plants. P1-9 was inferred to belong to the Basidiomycota based on the phylogenetic analysis of the 5.8S gene sequences but could not be identified to lower taxonomic levels. Further identification of the other four fungal clones to lower taxonomic levels was attempted based on phylogenetic analysis and sequence comparison of both the conserved 5.8S gene and the variable ITS regions. The origin of P2-6 was identified to be Glomerella and its anamorph Colletotrichum, the origins of P4-5 and P4-7 were Mycosphaerella and its anamorph Cladosporium, and the origin of P4-4 was the Herpotrichiellaceae. The direct approach to detection and taxonomic placement of endophytic fungi within host tissue without the need for conventional in vitro culturing is discussed. PMID- 11421645 TI - Evolution and biogeography of Alectryon (Sapindaceae). AB - Phylogenetic analysis of nucleotide sequences from four plastid loci (matK, partial trnK-matK introns, rps16 intron) and one nuclear locus (the internal transcribed spacer of rDNA; ITS-1) was conducted for 14 species of Alectryon and five related genera in Sapindaceae. Both matK and rps16 intron provide few informative characters within Alectryon, whereas ITS-1 provides the largest number of parsimony-informative characters and has the greatest sequence divergence between taxa. Support for branches in cladograms produced in PAUP increased markedly upon inclusion of ITS-1 data to matK and rps16 intron data. Analyses of each region alone or combined produced congruent results, suggesting that the regions are complementary. Phylogenetic analysis indicates that there are two main lineages within Alectryon, with A. subcinereus sister to the remaining sampled Alectryon taxa. Two morphological characters, presence/absence of petals and aril patterning, are congruent with the molecular phylogeny. One robustly supported clade is characterized by smooth arils and petals, in contrast to the taxa in the other major clade which have patterned arils and an absence of petals. These analyses also support a number of revised subgeneric groupings for Alectryon. The decision to submerge Heterodendrum in Alectryon is supported, although taxa belonging to Heterodendrum do not form a clade. The majority of the Australian Alectryon appear to belong to the tropical monsoonal/arid flora with species from both lineages being found in representative vine thickets across northern Australia. It appears that the seasonally dry rainforest communities comprise a number of elements that do not share common evolutionary histories within this genus. PMID- 11421646 TI - Higher-level phylogeny of new world vireos (aves: vireonidae) based on sequences of multiple mitochondrial DNA genes. AB - Interfamilial relationships of the New World songbird family Vireonidae are uncertain. Thus, we sequenced 3069 bp of four mitochondrial genes (cyt b, ND2, ND3, COI) from 19 taxa in five families and two outgroups, to examine higher level alliances with proposed relatives. We also sequenced cyt b and ND2 from an additional five vireonids to examine intergeneric relationships within the Vireonidae and incorporated 14 sequences of cyt b from GenBank to test the effects of taxon sampling on gene tree resolution. Families appeared monophyletic in all analyses, and the affinity of vireonids to Old World corvoids was corroborated. However, relationships among the Vireonidae and other families were not resolved. Sequences of vireonids revealed high levels of divergence within and between genera, with either Cyclarhis or Vireolanius positioned basally, depending on the analysis. On the basis of mitochondrial DNA and biogeographic evidence, vireonids represent a deep lineage derived from an Old World ancestor that colonized the New World, most likely via Beringia, with subsequent radiation in the Middle American tropics. We hypothesize postcolonization dispersal of the ancestor into Middle America, followed by extinction of the ancestor in North America. This extinction event left the North Temperate Zone unoccupied by any vireonid until northward reinvasion by some species of Vireo. Although the closest living relative of vireonids remains unidentified, broad-scale sequencing of additional extant corvoids with multiple molecular markers should further elucidate Old World alliances. PMID- 11421647 TI - A phylogenetic analysis of Doronicum (Asteraceae, Senecioneae) based on morphological, nuclear ribosomal (ITS), and chloroplast (trnL-F) evidence. AB - A phylogenetic analysis of the Old World genus Doronicum (26 species, 4 subspecies) based on sequence data of the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA, the chloroplast spacer trnL-F, and morphology is presented. Congruence among the three data sets was explored by the computing of several indices, all of which suggest homogeneity between only the two molecular matrices. We argue that the morphological data set contains poor phylogenetic signal and advocate simultaneous analysis of the three data sets (total evidence approach) so that morphological characters are tested for homology by congruence with molecular data. The resulting phylogenetic hypothesis allows several well-supported conclusions including the placement of a Corsican endemic (D. corsicum), sister to the remainder of the genus, and the inference that an early southern European or Mediterranean diversification took place in the genus. Shifts in morphological characters (e.g., homocarpy to heterocarpy) are confirmed to have evolved several times. Results from comparative studies of sequence data of the chloroplast gene ndhF support inclusion of Doronicum in tribe Senecioneae. PMID- 11421648 TI - Molecular phylogeny of the subfamilies in Geometridae (Geometroidea: Lepidoptera). AB - Molecular sequence data from three gene fragments were used to examine critically a provisional phylogenetic classification based on morphological characters of the Geometridae, one of the most species-rich families of moths. The sister group relationship between Geometridae and Drepanidae gained further support from the molecular analysis, which was based on the ND1 mitochondrial gene and the first and second expansion segments of the 28S ribosomal RNA gene. Although the alignment of the second expansion segment contained regions with many gaps, it provided the most resolution of the gene fragments. Parsimony analysis of the combined data resulted in a cladogram in which species belonging to Drepanidae, Larentiinae, and Sterrhinae formed monophyletic groups. The Ennominae did not form a monophyletic group but rather were contained within a broader monophyletic group including Archiearinae, Geometrinae, and Alsophilinae (represented by only one species per group in the present study). The molecular results were used to explore further the relationship between Sterrhinae and Larentiinae, the question as to whether Ennominae actually represent a monophyletic group, and the relationships between Ennominae and some of the other subfamilies. PMID- 11421649 TI - Molecular phylogenetics of the sexually deceptive orchid genus Ophrys (Orchidaceae) based on nuclear and chloroplast DNA sequences. AB - We present a phylogenetic analysis of the major lineages of the sexually deceptive orchid genus Ophrys based on nuclear ribosomal (nr) DNA (internal transcribed spacer region) and noncoding chloroplast (cp) DNA (trnL-trnF region) sequences. Sequence divergence within and among major Ophrys lineages was low for both nrDNA and cpDNA sequences. Separate analyses resulted in similar but poorly resolved trees. An incongruence length difference test revealed that nrDNA and cpDNA data sets were not incongruent. A combined analysis resulted in a better resolved phylogenetic hypothesis of relationships among the major Ophrys lineages. Our data strongly support a division of Ophrys into two groups. These groups do not correspond to the earlier proposed sections Euophrys and Pseudophrys and are thus in conflict with traditional classifications. Our results support a well-resolved monophyletic group that contains the geographically widespread O. bombyliflora, O. speculum, O. tenthredinifera, and the O. fusca-lutea lineage. Relationships in the other group are poorly resolved. Based on our observations that taxa with identical sequences at presumably rapidly evolving loci clearly differ in floral morphology, we hypothesize that the diversity in the genus Ophrys is the result of a recent radiation in this orchid lineage. PMID- 11421650 TI - Phylogenetic relationships inferred from ribosomal its sequences and biogeographic patterns in representatives of the genus Calopteryx (Insecta: Odonata) of the West Mediterranean and adjacent West European zone. AB - Western Europe is a reinvasion zone for the riverine dragonfly genus Calopteryx (Insecta: Odonata). Reinvasion may have been from central West Asia or from the West Mediterranean refugium. Phylogenetic relationships of West Mediterranean and West European taxa of the genus Calopteryx from different localities were inferred from sequences of the internal transcribed spacers (ITS1 and ITS2) of the nuclear ribosomal RNA genes. Twenty-six taxa belonging to the species groups C. splendens, C. meridionalis, C. haemorrhoidalis, C. virgo, C. xanthostoma, and C. exul were analyzed, with two North American species, C. amata and C. aequabilis, as outgroup. Sequence data and phylogenetic analyses were used to infer biogeographical patterns. The ribosomal spacers (ITS1 and ITS2) and the intervening 5.8S rDNA gene were amplified by PCR and sequenced. The ITS2 sequences of the West Mediterranean and West European calopterygids show no length variation but the ITS1 region was slightly variable in length. The sequence variation for ITS1 and ITS2 regions between different West Mediterranean and West European calopterygids was 14.5 and 6.1%, respectively. Phylogenetic relationships inferred from ITS sequences only partly confirm morphological data. A monophyletic origin of all West Mediterranean and West European species emerged. They are separated into two main clades; the splendens-like forms and the virgo/meridionalis/haemorrhoidalis group. Intraspecific variability, indicating different stages of speciation, was detected only in West Mediterranean representatives (e.g., C. xanthostoma) but not in invasive representatives in West Europe. The North African endemic C. exul is more closely related to the Italian C. s. caprai than to C. splendens sensu strictu. Based on the present information, Cretan populations are the only splendens-like taxa in addition to C. s. caprai that deserve subspecies status. PMID- 11421651 TI - The horizontal transfer of Agrobacterium rhizogenes genes and the evolution of the genus Nicotiana. AB - With the aim of understanding better the distribution and evolution of Agrobacterium rhizogenes genes transferred in the genus Nicotiana, 42 species were screened for presence of rolB, rolC, ORF13, and ORF14. The transferred sequences were then compared within the genus and with current bacterial sequences. The results obtained showed the presence of at least one bacterial gene in 15 species belonging to different subgenera. Sequence analyses supported the hypothesis of coevolution of bacterial and plant sequences, thus suggesting a possible role for the transferred genes in the early events of Nicotiana species differentiation. The high level of conservation of Agrobacterium sequences and the dependence of their expression from the plant physiological context along with previous data suggesting their involvement in the determination of the plant hormonal balance were all consistent with this hypothesis. The results are finally discussed also as to their relevance for the hypothesis of mono and multi ancient infection by Agrobacterium. PMID- 11421652 TI - Patterns of morphological, biochemical, and molecular evolution in the Oeneis chryxus complex (Lepidoptera: Satyridae): a test of historical biogeographical hypotheses. AB - Surveys of allozyme allele frequency and mitochondrial DNA (mtDNA) sequence variation were employed to test historical biogeographical hypotheses on the origin and unique distribution of the synchronized biennial, high-altitude butterflies of the Oeneis chryxus complex in western North America. Populations of O. c. stanislaus and O. ivallda from the central and northern Sierra Nevada are indistinguishable by use of allozyme allele frequency data, possessed nearly identical mtDNA cytochrome oxidase subunit 1 (COI) haplotypes, and were found to be relatively distantly related to O. c. chryxus from the Snake Range in eastern Nevada. However, individuals of O. ivallda from Piute Pass in the southern Sierra Nevada are more variable, with some individuals sharing mtDNA characteristics with O. c. chryxus. We find little support for the hypothesis proposed by W. Hovanitz in 1940 that O. c. stanislaus invaded the central Sierra Nevada from across the Great Basin and displaced O. ivallda, but cannot reject the hypothesis that ancestral Oeneis dispersed across the Great Basin to California. This result is congruent with hypotheses of dispersal across the Great Basin for the origin of some Sierran alpine organisms. PMID- 11421653 TI - Phylogeny of feather mite subfamily Avenzoariinae (Acari: Analgoidea: Avenzoariidae) inferred from combined analyses of molecular and morphological data. AB - Phylogenetic relationships among feather mites of the subfamily Avenzoariinae (Acari: Analgoidea: Avenzoariidae) were reconstructed by parsimony analysis of a combined data matrix. We analyzed 41 morphological characters and 246 molecular characters from a fragment of the 16S rDNA. Morphological trees were well supported at deep branches (genera and above), but showed much less support and resolution within genera. Molecular analyses produced trees with better resolution and support on terminal branches and worse support on basal branches. I(MF) index for the combined matrix pointed to the significant congruence of both data subsets with the whole of the data. The topology of the combined tree was close to the morphological tree in the deep branches and had well-resolved terminal branches as in the molecular tree. This suggests a considerable level of complimentarity between the two data sets. An analysis of association patterns of the mites and their hosts was conducted based on the results of the combined analyses for the Avenzoariinae and a phylogeny of their charadriiform hosts (compiled from various bird phylogeny hypotheses). The trees could be reconciled by the invoking of 12-13 cospeciation events, 6-7 duplications, 2 host shifts, and 26-29 sorting events. This suggests a high degree of cospeciation. PMID- 11421654 TI - Elongation factor-2: a useful gene for arthropod phylogenetics. AB - Robust resolution of controversial higher-level groupings within Arthropoda requires additional sources of characters. Toward this end, elongation factor-2 sequences (1899 nucleotides) were generated from 17 arthropod taxa (5 chelicerates, 6 crustaceans, 3 hexapods, 3 myriapods) plus an onychophoran and a tardigrade as outgroups. Likelihood and parsimony analyses of nucleotide and amino acid data sets consistently recovered Myriapoda and major chelicerate groups with high bootstrap support. Crustacea + Hexapoda (= Pancrustacea) was recovered with moderate support, whereas the conflicting group Myriapoda + Hexapoda (= Atelocerata) was never recovered and bootstrap values were always <5%. With additional nonarthropod sequences included, one indel supports monophyly of Tardigrada, Onychophora, and Arthropoda relative to molluscan, annelidan, and mammalian outgroups. New and previously published sequences from RNA polymerase II (1038 nucleotides) and elongation factor-1alpha (1092 nucleotides) were analyzed for the same taxa. A comparison of bootstrap values from the three genes analyzed separately revealed widely varying values for some clades, although there was never strong support for conflicting groups. In combined analyses, there was strong bootstrap support for the generally accepted clades Arachnida, Arthropoda, Euchelicerata, Hexapoda, and Pycnogonida, and for Chelicerata, Myriapoda, and Pancrustacea, whose monophyly is more controversial. Recovery of some additional groups was fairly robust to method of analysis but bootstrap values were not high; these included Pancrustacea + Chelicerata, Hexapoda + Cephalocarida + Remipedia, Cephalocarida + Remipedia, and Malaocostraca + Cirripedia. Atelocerata (= Myriapoda + Hexapoda) was never recovered. Elongation factor-2 is now the second protein-encoding, nuclear gene (in addition to RNA polymerase II) to support Pancrustacea over Atelocerata. Atelocerata is widely cited in morphology-based analyses, and the discrepancy between results derived from molecular and morphological data deserves greater attention. PMID- 11421655 TI - Historical biogeography and a mitochondrial DNA phylogeny of grouse and ptarmigan. AB - We sequenced 2690 nucleotides of mitochondrial DNA (mtDNA) including the entire control region (CR), partial 12S and 16S ribosomal RNAs, NADH dehydrogenase subunit 2, and cytochrome b genes from representatives of all the 17 living species of grouse and ptarmigan (Aves; Galliformes; subfamily Tetraoninae). Substitution rates and phylogenetic signals were variable among genes, with the CR being more informative than protein-coding and rRNA genes. Phylogenetic trees, computed with the CR or the concatenated sequences, indicate that: (1) genus Bonasa is monophyletic and basal within the subfamily, (2) all the other currently recognized genera of Tetraoninae are monophyletic, except Dendragapus; (3) D. obscurus is related to Centrocercus urophasianus and divergent from former D. canadensis and D. falcipennis, which, accordingly, may be ascribed to the distinct genus Falcipennis; (4) Tympanuchus, Dendragapus, and Centrocercus form a clade comprising taxa distributed exclusively in North America; and (5) the North American species of Bonasa (B. umbellus) and Lagopus (L. leucurus) are basal to their Eurasian and Holarctic congeneric species. These findings, and a dispersal vicariance analysis, support a North American origin of the subfamily and of all the genera of Tetraoninae, with the possible exception of Tetrao. Present species distributions might have been attained by at least three dispersal events from North America to Eurasia, involving the ancestors to Palearctic Bonasa, the ancestors to circumpolar Lagopus mutus/L. lagopus, and the clade leading to Tetrao/Falcipennis. According to a "standard calibration" of the mtDNA molecular clock (2% sequence divergence per million years), Bonasa split about 5-6 million years ago, the other genera diverged during the upper Pliocene, and most of the congeneric species with North American and Eurasian distributions (Bonasa, Lagopus, and Falcipennis) originated during the lower Pleistocene, well before the last interruption of the Beringian land bridge. PMID- 11421656 TI - Growth of arterioles precedes that of capillaries in stretch-induced angiogenesis in skeletal muscle. AB - Arteriolar growth accompanying capillary angiogenesis has been linked with hemodynamic factors resulting from increased blood flow. Here we describe the growth of arterioles occurring in rat skeletal muscles stretched by an overload due to the removal of agonist muscles, where blood flow was not increased, and we provide morphological evidence for the type of cells involved in this growth. Rat extensor digitorum longus (EDL) and extensor hallucis proprius (EHP) were overloaded by unilateral extirpation of their agonist, tibialis anterior. EDL muscles were taken for immunohistochemistry in cryostat sections to mark endothelial cells (Griffonia simplicifolia I, GSI lectin), smooth muscle cells and pericytes (alpha smooth muscle actin, alphaSMA), and "mature" arterioles (smooth muscle myosin heavy chains). EHP muscles were used for corresponding evaluation by confocal and electron microscopy. The number of capillaries surrounding muscle fibers was not significantly different after 1 week of stretch but was higher after 2 weeks (5.15 +/- 0.2 vs 4.3 +/- 0.2 in controls, P < 0.05). Similarly, capillary density (CD) and capillary/fiber ratio (C/F) gradually increased (CD 778 +/- 86 at 2 weeks vs 593 +/- 35 mm(-2) in controls, C/F 2.07 +/ 0.13 vs 1.38 +/- 0.06, respectively). In contrast, the number of alphaSMA positive vessels around fibers increased after 1 week (2.16 +/- 0.09 vs 0.25 +/- 0.02 in controls) and was lower after 2 weeks (1.42 +/- 0.24, P < 0.05, vs 1 week). Arteriolar density was higher at 1 (110.9 +/- 7.5 mm(-2)) and 2 weeks (70.7 +/- 12.1) with respect to controls (31.0 +/- 1.6 mm(-2)). The increased density was greater in alphaSMA-positive vessels <10 microm in diameter (controls 18.0 +/- 1.04, 1 week 77.2 +/- 4.5, 2 wk 42.2 +/- 9.0 mm(-2)) than in vessels >10 microm (13.0 +/- 0.8, 33.7 +/- 4.0, 29.5 +/- 4.7 mm(-2)). Electron microscopy showed "activated" (TEM fine structure) and proliferating (immunogold labeling for BrdU) fibroblasts in the vicinity of capillaries, some of which were embedded in the capillary basement membrane, consistent with a transformation into pericytes and possibly later smooth muscle cells. Confocal microscopy indicated that some mesenchymal cells became GSI positive and formed extended processes which contacted capillaries via tapered endings. Growth of arterioles in stretched muscles appears to involve proliferation of fibroblasts, which may migrate toward capillaries and precedes any apparent increase in capillarization. PMID- 11421657 TI - Preovulatory treatment of mice with anti-VEGF receptor 2 antibody inhibits angiogenesis in corpora lutea. AB - Adult mammalian angiogenesis occurs predominantly in female reproductive organs: the ovary and the uterus. Angiogenesis is very active during corpus luteum formation. A key regulator of angiogenesis is vascular endothelial growth factor (VEGF), which is highly expressed during corpus luteum formation. Inhibition of VEGF activity can block the formation and function of the corpora lutea by preventing angiogenesis. The VEGF receptor 2 (VEGF-R2) mediates the angiogenic action of VEGF and is expressed during corpus luteum formation. We hypothesized that treatment with an antibody against VEGF-R2 would inhibit luteal angiogenesis by blocking VEGF/VEGF-R2 interaction. Immature mice were induced to superovulate with PMSG/hCG resulting in neovascularization in the corpora lutea, as evidenced by abundant staining for the endothelial-specific adhesion molecule PECAM. Multiple doses of a monoclonal antibody against the VEGF-R2 (DC101) were administered to immature mice. Treatment was initiated 2 days prior to the induction of superovulation with PMSG/hCG. This antibody inhibited luteal angiogenesis as evidenced by the lack of PECAM staining in the center of the corpora lutea. Multiple dose treatment with antibody initiated prior to gonadotropin administration could not dissociate the luteal inhibition from the consequences of inhibition of angiogenesis in the developing follicle. Administration of a single, preovulatory dose of anti-VEGF-R2 antibody, such that follicular angiogenesis would not be affected, also inhibited luteal development, demonstrating that luteal angiogenesis is required for corpus luteal development. We conclude that VEGF acting through VEGF-R2 has an obligatory role in luteal angiogenesis and corpus luteum formation. PMID- 11421658 TI - Decorin inhibits endothelial migration and tube-like structure formation: role of thrombospondin-1. AB - Interactions between endothelial cell receptors and the extracellular matrix (ECM) play a critical, yet poorly understood role in angiogenesis. Based on the anti-adhesive role of decorin, we hypothesized that decorin binding to ECM molecules such as thrombospondin-1 (TSP-1) plays a regulatory role in endothelial tube-like structure (TLS) formation. To test this hypothesis, endothelial cells were plated on TSP-1, decorin, or mixed substrates of TSP-1 plus decorin. TLS formation was induced by applying type I collagen on the confluent endothelial monolayer. Cartilage decorin inhibited the formation of TLSs in a concentration dependent manner. On substrates of high decorin concentrations (2.5 and 5.0 microg/cm(2)) the reduction in TLSs was due either to a reduction in the number of adhering cells or to decreased cell migration. At low decorin concentrations (0.05 and 0.25 microg/cm(2)) the reduction in TLSs was independent of the number of attached cells. Time-lapse video microscopy revealed that decorin substrates facilitated homotypic aggregation and isolated cord formation at the expense of endothelial migration and TLS formation. Consistent with the reduced migration, endothelial cells formed fewer vinculin-positive focal adhesions and actin-stress fibers on decorin substrates. Endothelial migration and TLS formation were also significantly inhibited by skin decorin and the protein core of cartilage decorin. The inhibition of TLS formation by the protein core of cartilage decorin was potentiated by TSP-1. These findings suggest that decorin alone or in combination with TSP-1 interferes with the activation of endothelial cell receptors by ECM molecules, thus blocking intracellular signals that induce cytoskeletal reorganization, migration, and TLS formation. PMID- 11421659 TI - Protective role of an endothelin-converting enzyme inhibitor (FR901533) in hepatic ischemia/reperfusion injury. AB - INTRODUCTION: There is evidence that endothelin (ET) contributes to disturbances of the hepatic microcirculation after warm ischemia/reperfusion (I/R) by causing vasoconstriction and by enhancing leukocyte endothelium interactions. The aim of this study was to investigate a possible protective role of the endothelin converting enzyme (ECE) inhibitor FR901533 in this setting. METHODS: In an in vivo model (42 Wistar rats), hepatic ischemia was induced for 30 min by Pringle's maneuver. Sham operated (I), untreated ischemic (II), and treatment (III) groups with FR901533 (1 mg/kg bw iv) were investigated. The effect of FR901533 in I/R was assessed by in vivo microscopy (30-90 min after reperfusion), measurement of local tissue pO2 (30 and 60 min after reperfusion), and determination of AST/ALT levels (2 h, 6 h, and 2, 6, and 14 days after reperfusion). RESULTS: In the untreated ischemic group (II) sinusoidal constriction to 76.3 +/- 4.2% of basic diameters was observed, leading to significant decreases in perfusion rate (82.3 +/- 3.6% of sham group) and in liver tissue pO(2) (43.5 +/- 3.2% of sham group) (P < 0.05). In addition, we found an increased percentage of stagnant leukocytes in sinusoids (138.3 +/- 9.8) and sticking leukocytes in postsinusoidal venules (155.2 +/- 3.3% of sham group) (P < 0.05). Hepatocellular damage (AST/ALT increase to 430.6 +/- 47.7 U/L/200.2 +/- 23.8 U/L, pre: 27.4 +/- 2.7 U/L/28.1 +/- 2.7 U/L) was detected 6 h after reperfusion (P < 0.05). Administration of the ECE inhibitor before ischemia significantly reduced I/R injury. Sinusoidal diameters were maintained (102.2 +/- 1.7%), while perfusion rate (93.1 +/- 1.8%) and tissue pO2 (105.3 +/- 2.7%) increased significantly (P < 0.05). Hepatocellular damage was decreased (AST/ALT levels after 6 h of reperfusion: 166.6 +/- 26.3 U/L/132.4 +/- 22.5 U/L, P < 0.05) and leukocyte sticking and rolling were significantly reduced (P < 0.05). CONCLUSION: Our results provide evidence that the new therapeutic approach with an ECE inhibitor is effective in reducing hepatic I/R injury. PMID- 11421660 TI - Measurement of membrane potential and intracellular Ca(2+) of arteriolar endothelium and smooth muscle in vivo. AB - We have developed an intensity analysis technique for fluorescence microscopy that allows us to measure, in real time, the diameter and the membrane potential or intracellular calcium ([Ca(2+)]i) of in vivo arteriolar endothelium or smooth muscle. Cheek pouch arterioles of anesthetized hamsters were luminally or abluminally labeled with Di-8-ANEPPS, a voltage-sensitive dye, or Fura PE3, a calcium indicator. The peak fluorescence intensities of the images were used to locate the endothelium or smooth muscle. The changes in membrane potential or [Ca(2+)]i were determined based on the ratiometric analysis of fluorescence intensity of the endothelium or smooth muscle. Membrane depolarization of the smooth muscle using KCl caused a decrease in the ratio of emission, 620 nm/560 nm ( approximately 6 mV/% ratio). The ratio of excitation, 340 nm/380 nm, increased with increasing free Ca(2+). Methacholine, a muscarinic receptor agonist, caused arteriolar dilation (12.2 +/- 0.9 µm). It produced hyperpolarization of the endothelium and smooth muscle (2.8 +/- 0.6% and 2.3 +/- 0.3% in ratio). Methacholine also induced an increase in [Ca(2+)]i (11.0 +/- 1.1% in ratio) of the endothelium. In contrast, methacholine caused a biphasic change in [Ca(2+)]i of the smooth muscle, a rapid reduction (-3.4 +/- 0.2% in ratio) followed by a prolonged increase (2.4 +/- 0.2% in ratio). These results demonstrate that the peak intensity analysis can be used to determine in real time the changes in membrane potential or [Ca(2+)]i of in vivo endothelium or smooth muscle. PMID- 11421661 TI - Mathematical analysis of hemoglobin spectrophotometry in microvessels. AB - Spectrophotometry of hemoglobin in microvessels is commonly performed by collecting light either from a small region around the vessel centerline or from the entire lumen of the vessel. In the latter instance, parallel rays of light may not encounter the same amount of absorbing species. Hence, a phenomenon similar to the sieve effect reported in the literature on hemoglobin spectrophotometry may be expected to occur. Although it has been observed that under such circumstances nonlinearities in calibration characteristics arise, the implications of this effect on the interpretation of the spectrophotometric mean concentration have never been addressed so far. Mathematical analysis of hemoglobin spectrophotometry in microvessels, performed in this study, reveals that for practical situations the calibration curve is indeed nonlinear. Moreover, the spectrophotometric mean oxygen saturation is an overestimate of the mean oxygen saturation during oxygenation and an underestimate of the mean oxygen saturation during deoxygenation. These deviations depend upon the manner in which the total heme concentration is distributed within the lumen. Application of the analysis to artificial microvessels showed that the observed superior oxygen transport characteristics of flowing erythrocyte suspensions and hemoglobin solution mixtures could in part be due to the assumptions underlying the procedure used to interpret the experimental results. The implications of this result on models for oxygen transport in microvessels are discussed along with possible resolutions. PMID- 11421662 TI - Laser-Doppler imaging of forearm skin: perfusion features and dependence of the biological zero on heat-induced hyperemia. PMID- 11421663 TI - Influence of mast cells on leukocyte-independent plasma extravasation during endotoxemia. PMID- 11421664 TI - In situ Fe K-edge X-ray absorption spectroscopy of a nitrosyl iron(II) porphyrin adduct adsorbed on a high-area carbon electrode in aqueous electrolytes. PMID- 11421666 TI - The first triple-layer 2-D coordination polymer:. PMID- 11421665 TI - Proton mediated partial degradation of closo-dicarbaboranes. PMID- 11421667 TI - Bent and stretched but not yet to the breaking point: C(8)-C(16) sp carbon chains that span two platinum atoms and the first structurally characterized 1,3,5,7,9,11,13,15-octayne. PMID- 11421668 TI - Composition and structure of the anion layer in the organic superconductor kappa' (ET)(2)Cu(2)(CN)(3): optical study. PMID- 11421669 TI - An axial phosphine diadduct of diruthenium(II,III) tetraacetate. PMID- 11421670 TI - Protonation of platinated adenine nucleobases. Gas phase vs condensed phase picture. AB - Protonation of adenine carrying a Pt(II) moiety either at N7, N3, or N1 is possible in solution, but the site of protonation is influenced by the location of the Pt(II) electrophile and to some extent also by the overall charge of the metal entity (+2, +1, 0, -1), hence the other ligands (NH(3), Cl(-), OH(-)) bound to Pt(II). Quantum chemical calculations based on density functional theory (DFT) have been carried out for intrinsic protonation energies of adenine complexes carrying the following Pt(II) species at either of the three ring N atoms: [Pt(NH(3))(3)](2+) (1), trans- [Pt(NH(3))(2)Cl](+) (2a), cis-[Pt(NH(3))(2)Cl](+) (2b), trans-[Pt(NH(3))(2)Cl(2)] (3a), cis-[Pt(NH(3))Cl(2)] (3b), [PtCl(3)](-) (4), trans-[Pt(NH(3))(2)OH](+) (5a), cis-[Pt(NH(3))(2)(OH)](+) (5b), trans [Pt(NH(3))(OH)(2)] (6a), cis-[Pt(NH(3))(OH)(2)] (6b), and [Pt(OH)(3)](-) (7). The data have been compared with results derived from solution studies (water) and X ray crystallography, whenever available. The electrostatic effects associated with the charge of the metal entity have the major influence on the calculated intrinsic (gas phase) proton affinities, unlike the condensed phase data. Nevertheless, the relative gas phase trends correlate surprisingly well with condensed phase data; i.e., variation of the pK(a) values measured in solution is consistent with the calculated gas phase protonation energies. In addition to a systematic study of the ring proton affinities, proton transfer processes within the platinated adenine species were often observed when investigating Pt adducts with OH(-) ligands, and they are discussed in more detail. To the best of our knowledge, this is the first study attempting to find a systematic correlation between gas phase and condensed phase data on protonation of metalated nucleobases. The gas phase data provide a very useful complement to the condensed phase and X-ray experiments, showing that the gas phase studies are capable of valuable predictions and contribute to our understanding of the solvent and counterion effects on metal-assisted proton shift processes. PMID- 11421671 TI - A combined quantum mechanical and statistical mechanical study of the equilibrium of trimethylaluminum (TMA) and oligomers of (AlOCH(3))(n) found in methylaluminoxane (MAO) solution. AB - Density Functional Theory (DFT) has been used to calculate the energies of over 30 different structures with the general formula (AlOMe)(n).(AlMe(3))(m) where n ranges from 6 to 13 and m ranges between 1 and 4, depending upon the structure of the parent (AlOMe)(n) cage. The way in which TMA (trimethylaluminum) bonds to MAO (methylaluminoxane) has been determined as well as the location of the acidic sites present in MAO caged structures. Topological arguments have been used to show that TMA does not bind to MAO cages where n = 12 or n > or = 14. The ADF energies in conjunction with frequency calculations based on molecular mechanics have been used to estimate the finite temperature enthalpies, entropies, and free energies of the TMA containing MAO structures. Using the Gibbs free energies found for pure MAO structures calculated in a previous work, in conjunction with the free energies of TMA containing MAO structures obtained in the present study, it was possible to determine the percent abundance of each TMA containing MAO within the temperature range of 198.15 K-598.15 K. We have found that very little TMA is actually bound to MAO. The Me/Al ratio on the MAO cages is determined as being approximately 1.00, 1.01, 1.02, and 1.03 at 198, 298, 398, and 598 K, respectively. Moreover, the percentage of Al found as TMA has been calculated as being 0.21%, 0.62%, 1.05%, and 1.76% and the average unit formulas of (AlOMe)(18.08).(TMA)(0.04), (AlOMe)(17.04).(TMA)(0.11), (AlOMe)(15.72).(TMA)(0.17), and (AlOMe)(14.62).(TMA)(0.26) have been determined at the aforementioned temperatures. PMID- 11421672 TI - Fluxional behavior of the dinitrogen ligand 2,9-dimethyl-1,10-phenanthroline in cationic methyl platinum(II) complexes. AB - The ionic methylplatinum(II) complexes [Pt(Me)(L)(dmphen)]X (dmphen = 2,9 dimethyl-1,10-phenanthroline, L = Me(2)SO, X = PF(6)(-) 1a, BF(4)(-) 1b, CF(3)SO(3)(-) 1c, ClO(4)(-) 1d, B(C(6)H(5))(4)(-) 1e, [B(3,5 (CF(3))(2)C(6)H(3))(4)](-) 1f; L = n-Bu(2)SO, X = CF(3)SO(3)(-) 1g; L = PPh(3), X = PF(6)(-) 2a, BF(4)(-) 2b, CF(3)SO(3)(-) 2c, ClO(4)(-) 2d, B(C(6)H(5))(4)(-) 2e, [B(3,5-(CF(3))(2)C(6)H(3))(4)](-) 2f; X = CF(3)SO(3)(-), L = CyNH(2) 3a, i PrNH(2) 3b, 2,6-Me(2)py 3c, EtNH(2) 3d, AsPh(3) 3e, dimethylthiourea (Me(2)th) 3f and the uncharged [Pt(Me)(X)(dmphen)] (X = SCN(-) 4a, SeCN(-) 4b) complexes have been synthesized and fully characterized. In chloroform, as well as in acetone or methanol, complexes 1a-1g, 2a-2h (X = Cl(-) g, NO(2)(-) h, formed "in situ"), and 3e show dynamic behavior due to the oscillation of the symmetric chelating ligand dmphen between nonequivalent bidentate modes. All the other compounds feature a static structure in solution. The crystal structure of 2a shows a tetrahedral distortion of the square planar coordination geometry, a loss of planarity of the dmphen ligand, and, most notably, a rotation of the dmphen moiety, around the N1 N2 vector, to form a dihedral angle of 42.64(8) degrees with the mean coordination plane. The hexafluorophosphate ion lies on the side of the phenanthroline ligand. The interionic structures of 2a, 2b, and 2f were investigated in CDCl(3) at low temperature by (1)H-NOESY and (19)F[(1)H]-HOESY NMR spectroscopies. Whereas PF(6)(-) (2a) and BF(4)(-) (2b) show strong contacts with the cation [Pt(Me)(PPh(3))(dmphen)](+), being located preferentially on the side of the phenanthroline ligand, the [B(3,5-(CF(3))(2)C(6)H(3))(4)](-) (2f) ion does not form a tight ion pair. The dynamic process was studied by variable temperature NMR spectroscopy for 1a-1f and 2a-2h in CDCl(3). The activation energies DeltaG(298) for the sulfoxide complexes 1a-1f are lower than those of the corresponding phosphine complexes 2a-2f by approximately 10 kJ mol(-)(1). The nature of the counteranion exerts a tangible influence on the fluxionality of dmphen in both series of complexes 1 and 2. The sequence of energies observed for 2a-2h encompasses an overall difference of about 16 kJ mol(-)(1), increasing in the order Cl(-) approximately NO(2)(-) << CF(3)SO(3)(-) < ClO(4)(-) < B(C(6)H(5))(4)(-) < BF(4)(-) approximately PF(6)(-) < B(3,5 (CF(3))(2)C(6)H(3))(4)(-). Acetone and methanol have an accelerating effect on the flipping. Concentration-dependent measurements, carried out in CDCl(3) for 2a with n-Bu(4)NPF(6) and the ligands dmphen, n-Bu(2)SO, sec-Bu(2)SO, and sec-Bu(2)S showed that the rate of the fluxional motion is unaffected by added n Bu(4)NPF(6), whereas in the other cases this increases linearly with increasing ligand concentration, according to a pattern of behavior typical of substitution reactions. Dissociative and associative mechanisms can be envisaged for the observed process of flipping. Dissociation can be prevalent within the ion pair formed by a "noncoordinating" anion with the metallic cationic complex in chloroform. Among the possible associative mechanisms, promoted by polar solvents or by relatively strong nucleophiles, a consecutive displacement mechanism is preferred to intramolecular rearrangements of five-coordinate intermediates. PMID- 11421673 TI - Structural, computational, and (59)Co NMR studies of primary and secondary amine complexes of Co(III) porphyrins. AB - Four novel low-spin bis(amine) Co(III) porphyrins [Co(TPP)(BzNH(2))(2)](SbF(6)), 1, [Co(TPP)(1-BuNH(2))(2)](SbF(6)), 2, [Co(TPP)(PhCH(2)CH(2)NH(2))(2)](SbF(6)), 3, and [Co(TPP)(1-MePipz)(2)](SbF(6)), 4, have been synthesized and characterized by low-temperature X-ray crystallography, IR, electronic, and NMR ((1)H, (13)C, and (59)Co) spectroscopy. The mean Co-N(p) distance for the four structures is 1.986(1) A. The Co-N(ax) distances for the 1 degrees amine derivatives average to 1.980(5) A; the axial bonds of the 2 degrees amine derivative are significantly longer, averaging 2.040(1) A. The porphyrin core conformation of 4 is significantly nonplanar (mixture of S(4)-ruf and D(2d)-sad distortions) due to a staggered arrangement of the axial ligands over the porphyrin core and meso phenyl group orientations < 90 degrees. The X-ray structures have been used with the coordinates for [Co(TPP)(Pip)(2)](NO(3)) (Scheidt et al. J. Am. Chem. Soc. 1973, 95, 8289-8294.) to parametrize a molecular mechanics (MM) force field for bis(amine) complexes of Co(III) porphyrins. The calculations show that two types of crystal packing interactions (van der Waals and hydrogen bonding) largely control the crystallographically observed conformations. Gas phase conformational energy surfaces have been computed for these complexes by dihedral angle driving methods and augmented with population distributions calculated by MD simulations at 298 K; the calculations demonstrate that the bis(1 degrees amine) complexes are significantly more flexible than the bis(2 degrees amine) analogues. (59)Co NMR spectra have been acquired for a range of [Co(TPP)(amine)(2)]Cl derivatives as a function of temperature. The (59)Co chemical shifts increase linearly with increasing temperature due to population of thermally excited vibrational levels of the (1)A(1) ground state. Activation energies for molecular reorientation (tumbling) have been determined from an analysis of the (59)Co NMR line widths as a function of 1/T; lower barriers exist for the conformationally rigid 2 degrees amine derivatives (2.6-3.8 kJ mol(-1)). The (59)Co chemical shifts vary linearly with the DFT-calculated radial expectation values (3d) for the Co(III) ion. The correlation leads to the following order for the sigma-donor strengths of the axial ligands: BzNH(2) > or = Cl(-) > 1-BuNH(2) > PhCH(2)CH(2)NH(2) > 1 Bu(2)NH > Et(2)NH. The (59)Co NMR line widths are proportional to the square of the DFT-calculated valence electric field gradient at the Co nucleus. Importantly, this is the first computational rationalization of the (59)Co NMR spectra of Co(III) porphyrins. PMID- 11421674 TI - Polynuclear polypyridine complexes incorporating Ru(II), Os(II), and Pt(II): decanuclear dendrimeric antennas. AB - The "complexes as metal/complexes as ligand" strategy has been used to prepare new polynuclear heterometallic complexes containing two, 3, 4, and ten metal units. The metal ions employed are Ru(2+), Os(2+), and Pt(2+); the bridging ligands are 2,3- and 2,5-dpp (dpp = bis(2-pyridyl)pyrazine), and the terminal ligands are Cl(-) and bpy (bpy = 2,2'-bipyridine). [Os(bpy)(2)(mu-2,3-dpp)Ru[(mu 2,3-dpp)PtCl(2)](2)](PF(6))(4) (OsRuPt(2)) and [Os[(mu-2,3-dpp)Ru[(mu-2,3 dpp)PtCl(2)](2)](3)](PF(6))(8) (OsRu(3)Pt(6)), which have a dendritic structure, are the first examples of polynuclear complexes based on the dpp bridging ligand simultaneously containing three different metals. All of the new compounds feature absorption spectra with intense ligand-centered transitions in the UV region and metal-to-ligand charge transfer (MLCT) bands in the visible region. The complexes are luminescent both at room temperature in acetonitrile solution and at 77 K in butyronitrile rigid matrix. OsRuPt(2) and OsRu(3)Pt(6) luminesce in the near-infrared spectral region at 975 and 875 nm, respectively, at room temperature. This indicates that the emitting state is a (formally triplet) MLCT involving Os(2+) and the bridging ligand mu-2,3-dpp and that all the absorbed energy is efficiently channeled to the single Os-based unit. OsRu(3)Pt(6) appears to be the first decanuclear species displaying such efficient antenna effect. PMID- 11421675 TI - Synthesis, characterization, Langmuir-Blodgett film-forming properties, and second-harmonic-generation studies of ruthenium(II) complexes with long hydrocarbon chains. AB - Several ruthenium(II) surfactants of general formula [Ru(bpy)(2)L](2+) [L = N-(2 pyridylmethylene)-3,4-bis(tetradecyloxy)benzenamine, N-(2-pyridylmethylene)-3,4 bis(octadecyloxy)benzenamine, 9-(3,4-bis(tetradecyloxy)benzenamino)-4,5 diazafluorene, 9-(4-(tetradecyloxy)-4'-azabenzenamino)-4,5-diazafluorene, 4,7 dinonadecyl-1,10-phenanthroline] and [Ru(L)(3)](2+) (L = 4,7-dinonadecyl-1,10 phenanthroline) were synthesized and characterized by elemental analysis, (1)H NMR, UV-vis, luminescence spectroscopy, and cyclic voltammetry. Their Langmuir Blodgett (LB) film formation properties and surface pressure-area (pi-A) isotherms were also studied. Two complexes were found to show second-harmonic generation (SHG) behavior. PMID- 11421676 TI - Ab initio structure determination of a small-pore framework sodium stannosilicate. AB - The structure of a small-pore framework sodium stannosilicate Na(2)SnSi(3)O(9).2H(2)O (AV-10) has been determined ab initio from powder X-ray diffraction data (XRD). The unit cell is orthorhombic (space group C222(1), Z = 4) with cell dimensions a = 7.9453(5), b = 10.3439(7), c = 11.6252(7) A, V = 955 A(3). The structure of AV-10 is composed of corner sharing SnO(6) octahedra and SiO(4) tetrahedra, forming a three-dimensional framework structure. The SiO(4) tetrahedra form helix chains along [001] interconnected by SnO(6) octahedra. The SnO(6) octahedra are isolated by SiO(4) tetrahedra and, thus, there are no Sn-O Sn linkages. AV-10 has been characterized by chemical analysis, powder XRD, scanning electron microscopy, (29)Si, (119)Sn, single- and (FAM) triple-quantum (23)Na MAS NMR spectroscopy, thermogravimetry (TGA), and nitrogen adsorption isotherms. The zeolitic water of AV-10 is reversibly lost. The dehydrated material has been studied in situ by powder XRD, TGA, and, in particular, triple quantum (23)Na MAS NMR. PMID- 11421677 TI - Autoxidation of substituted phenols catalyzed by cobalt Schiff base complexes in supercritical carbon dioxide. AB - This first study of O(2) oxidation (autoxidation) of substituted phenols catalyzed by a dioxygen carrier in supercritical carbon dioxide (scCO(2)) provides additional insights into the established mechanism of reactions that have been much studied in conventional solvents. As has been long believed, the cobalt(II) dioxygen carriers of the class represented by [[N,N'-bis(3,5-di-tert butylsalicylidene)-1,2-cyclohexanediaminato(2-)]cobalt(II)], Co(salen), show both oxidase and oxygenase activities during oxygenation of substituted phenols in scCO(2). The catalytic autoxidation of 2,6-di-tert-butylphenol (DTBP) and 3,5-di tert-butylphenol (35-DTBP) in scCO(2) was studied by analysis of products in batch reactions with carefully controlled variables, in the presence of a large excess of O(2), at 207 bar of total pressure and a reaction temperature of 70 degrees C. The oxidation of 35-DTBP yielded only traces of products under the same experimental conditions that converted DTBP totally to a mixture of the oxygenation product 2,6-di-tert-butyl-1,4-benzoquinone (DTBQ) and the related product of radical coupling 3,5,3',5'-tetra-tert-butyl-4,4'-diphenoquinone (TTDBQ). The effects on conversion of DTBP to products and on selectivity between the two products were studied for variations in temperature and the concentrations of catalyst, oxygen, and methylimidazole. Selectivity in favor of the O-transfer product DTBQ over the self-coupling of the phenoxy radical was observed upon changing the oxygen concentration. In contrast, selectivity remained unaffected over a wide range of temperatures and catalyst concentrations. The oxygen dependence of both the conversion and selectivity showed saturation effects identifying the dioxygen complex as the effective oxidant in both the initial radical formation step and the oxygenation of that radical. No direct reaction is observed between the electrophilic phenoxy radical and O(2). PMID- 11421678 TI - Bonding analysis of inorganic transition-metal cubic clusters. 6. Copper(I) dithiolato species and related compounds. AB - Extended Huckel and density functional calculations carried out on 128-MVE Cu(8)(dithiolato)(6) edge-bridged cubic clusters indicate that their stability is mainly driven by the chelating effect of the ligands, which provide a stable 16 electron configuration to the approximately trigonal planar metal centers. Nevertheless, a weak but significant d(10)-d(10) bonding interaction is present which is rather independent from the dithiolato bite effect. The metal centers have a nonbonding 4p(z)() vacant AO pointing to the center of the cube available for bonding to an encapsulated atom. The electronic closed-shell requirement is satisfied for the 136-MVE and 140-MVE counts, respectively, when a main-group atom or a transition-metal atom is incorporated in the middle of the cube. The bonding within these dithiolato compounds is compared to other edge-bridged M(8) cubic clusters. In particular, it is shown that clusters of higher nuclearity but containing an M(8) cubic core are related to the dithiolato species. Indeed, their outer metal atoms can be considered as belonging to the ligand shell, interacting with the M(8) cube in a way similar to the dithiolato ligands in the Cu(8) species. PMID- 11421679 TI - Structure and electron affinity of platinum fluorides. AB - The structure, stability, and electron affinity of the even numbered molecular platinum fluorides PtF(2)(n) (n = 1-4) were studied by scalar relativistic density functional and coupled cluster methods. The di, tetra, and hexafluorides possess triplet ground states, while PtF(8) is a singlet. Formation of the latter from PtF(6) and F(2) is found to be endothermic. Differences between adiabiatic and vertical electron affinities are only significant for PtF(2). PMID- 11421680 TI - Exchange pathways in tris-mu-thiolato-bridged [Cr(2)L(3)](ClO(4)2Cl*H2O*MeOH (L=2,6-bis(aminomethyl)-4-tert-butyl-thiophenolate). AB - From polarized optical absorption and emission spectra of the tris-mu-thiolato bridged [Cr(2)L(3)](ClO(4))(2)Cl.H(2)O.MeOH (L = 2,6-bis(aminomethyl)-4-tert butyl-thiophenolate) 1 in the visible and near UV, the exchange splittings of the (4)A(2) ground and the (2)E and (2)T(1) excited states are determined. In view of the large Cr-Cr distance of 3.01 A, the antiferromagnetic ground state splitting with J = 78 cm(-1) (H = J(S(A).S(B))) is large compared to other triply bridged Cr(3+) dimers. This can be rationalized using a model based on a valence bond approach, in which the exchange splittings are derived from configuration interactions between the ground electron configuration and ligand-to-metal (LMCT) as well as metal-to-metal (MMCT) charge-transfer configurations. It allows us to distinguish the interactions via ligand orbitals from the direct interactions between the metal-centered orbitals. We compare our results with those obtained for the tri-mu-hydroxo-bridged [Cr(2)(OH)(3)(tmtame)(2)](NO(3))(3) (tmtame = N,N',N' '-trimethyl-1,1,1-tris(aminomethyl)ethane) 2. In the latter the interactions via the ligands are negligible, whereas they play an important role in 1, due to the softness of the sulfur ligand atoms. PMID- 11421681 TI - An X-ray study of the effect of the bite angle of chelating ligands on the geometry of palladium(allyl) complexes: implications for the regioselectivity in the allylic alkylation. AB - X-ray crystal structures of a series of cationic (P-P)palladium(1,1 (CH(3))(2)C(3)H(3)) complexes (P-P = dppe (1,2-bis(diphenylphosphino)ethane), dppf (1,1'-bis(diphenylphosphino)ferrocene), and DPEphos (2,2' bis(diphenylphosphino)diphenyl ether)) and the (Xantphos)Pd(C(3)H(5))BF(4) (Xantphos = 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene) complex have been determined. In the solid state structure, the phenyl rings of the ligand are oriented in the direction of the nonsymmetrically bound [1,1-(CH(3))(2)C(3)H(3)] moiety. An increase of the bite angle of the chelating ligand results in an increase of the cone angle. In complexes containing ligands having a large cone angle, the distances between the phenyl rings and the allyl moiety become small, resulting in a distortion of the symmetry of the palladium-allyl bond. In solution, two types of dynamic exchange have been observed, the pi-sigma rearrangement and the apparent rotation of the allyl moiety. At the same time, the folded structure of the ligand changes from an endo to an exo orientation or vice versa. The regioselectivity in the palladium-catalyzed allylic alkylation of 3-methyl-but-2-enyl acetate is determined by the cone angle of the bidentate phosphine ligand. Nucleophilic attack by a malonate anion takes place preferentially at the allylic carbon atom having the largest distance to palladium. Ligands with a larger cone angle direct the regioselectivity to the formation of the branched product, from 8% for dppe (1) to 61% found for Xantphos (6). The influence of the cone angle on the regioselectivity has been assigned to a sterically induced electronic effect. PMID- 11421682 TI - Polyhedral-based nonlinear optical materials. 2. Theoretical investigation of some new high nonlinear optical response compounds involving polyhedral bridges with charged aromatic donors and acceptors. AB - A theoretical study of several new classes of polyhedral-based molecules has shown that these species display large calculated nonlinear optical responses. These new classes of molecules are based on charged aromatic subunits connected through polyhedral cluster bridges, such as closo-[1-(C(7)H(6))-12 (C(5)Me(4))C(2)B(10)H(10)]. These compounds show calculated first hyperpolarizabilities (beta) ranging from 6.5 to 8413.9 x 10(-30) cm(5) esu(-1). A basis for understanding the origin of these large responses is proposed based on the two-state model and consideration of the orbital and electronic features of the molecules. In general, the highest occupied molecular orbitals for these species are localized on the aromatic donor rings, such as the cyclopentadienyl system, while the lowest unoccupied molecular orbitals are largely on the aromatic acceptor rings, such as the tropylium system. The electronic properties of these polyhedral-based systems appear to be significantly different from the analogous organic [5.6.7]quinarene system (tropyliumcyclopentadienylbenzene). The organic quinarene appears to behave as a completely electron-delocalized system over all three rings while the polyhedral-based compounds can best be described as consisting of two relatively independent, highly polarized regions. PMID- 11421683 TI - Polyhedral-based nonlinear optical materials. 3. Synthetic studies of cyclopentadiene- and cycloheptatriene-substituted polyhedral compounds: synthesis of 1,12-[C(7)H(7)C(2)B(10)H(10)(C(5)H(3)Me(2))] and related species. AB - The synthesis and characterization of a series of new olefin-substituted closo 1,12-[C(2)B(10)H(12)] compounds is described. The reaction of deprotonated closo 1,12-dicarbadodecaborane with 7-methoxycycloheptatriene yields the cycloheptatriene-substituted carborane compound, closo-[1-(1-C(7)H(7))-12-(H) C(2)B(10)H(10)] (3). Deprotonation of 3 with butyllithium and subsequent reaction with 3,4-dimethyl-2-cyclopenten-1-one was found to yield the bis-olefin substituted carbinol compound [1-(1-C(7)H(7))-12-(C(5)H(4)-1-(OH)-3,4-(CH(3))(2)) C(2)B(10)H(10)] (5) in good yield. Acidic dehydration of 5 quantitatively converted it into the simple bis-olefin cage compound [1-(1-C(7)H(7))-12 (C(5)H(3)-3,4-(CH(3))(2))-C(2)B(10)H(10)] (6). Finally, thermal treatment of 6 in refluxing toluene was employed to prepare the isomerized cycloheptatriene compound [1-(4-C(7)H(7))-12-(C(5)H(3)-3,4-(CH(3))(2))-C(2)B(10)H(10)] (7) in high yield. These compounds represent the first metal-free cyclopentadiene and bis olefin large cluster species reported in which the C(5) ring is attached directly to the cage. The complete characterization of products by multinuclear NMR ((1)H, (11)B, and (13)C), infrared, UV-visible, and mass spectroscopic analyses is presented. The X-ray crystal structures of 3 and 7 are also reported. X-ray data for 3: triclinic system, space group P&onemacr; with cell constants a = 6.6807(3) A, b = 10.2939(3) A, c = 10.3962(4) A, alpha = 89.342(2) degrees, beta = 74.610(2) degrees, gamma = 83.373(2) degrees, Z = 2, R1 = 0.0487 (wR2 (all data) = 0.1272). X-ray data for 7: monoclinic system, space group P2(1)/c with cell constants a = 7.7207(7) A, b = 15.8730(14) A, c = 15.5493(13) A, beta = 99.146(2) degrees, Z = 4, R1 = 0.0761 (wR2 (all data) = 0.2050). PMID- 11421684 TI - Coordination trends in alkali metal crown ether uranyl halide complexes: the series [A(crown)]2[UO(2)X(4)] where A=Li, Na, K and X=Cl, Br. AB - UO(2)(C(2)H(3)O(2))(2).2H(2)O reacts with AX or A(C(2)H(3)O(2) or ClO(4)) (where A = Li, Na, K; X = Cl, Br) and crown ethers in HCl or HBr aqueous solutions to give the sandwich-type compounds [K(18-crown-6)](2)[UO(2)Cl(4)] (1), [K(18-crown 6)](2)[UO(2)Br(4)] (2), [Na(15-crown-5)](2)[UO(2)Cl(4)] (3), [Na(15-crown 5)](2)[UO(2)Br(4)] (4), [Li(12-crown-4)](2)[UO(2)Cl(4)] (5), and [Li(12-crown 4)](2)[UO(2)Br(4)] (6). The compounds have been characterized by single-crystal X ray diffraction, powder diffraction, elemental analysis, IR, and Raman spectroscopy. The [UO(2)X(4)](2-) ions coordinate to two [A(crown)](+) cations through the four halides only (2), through two halides only (3), through the two uranyl oxygens and two halides (3, 4), or through the two uranyl oxygen atoms only (5, 6). Raman spectra reveal nu(U-O) values that correlate with expected trends. The structural trends are discussed within the context of classical principles of hard-soft acid-base theory. PMID- 11421685 TI - Self-assembly of a porphyrin array via the molecular recognition approach: synthesis and properties of a cyclic zinc(II) porphyrin trimer based on coordination and hydrogen bonding. AB - Self-assembly of [5-(pyrazol-4-yl)-10,20-bis(p-tolyl)-15- (2 ethoxycarbonylphenyl)porphyrinato]-zinc(II) (1), designed to have both a coordination site and a hydrogen bonding site, leads to a stable cyclic trimer array where coordination of the pyrazole nitrogen to the zinc(II) ion as well as hydrogen bonding between carbonyl oxygen and pyrazole NH holds each zinc(II) porphyrin. The recognition event for pyrazole has been confirmed preliminarily in the model studies using [5-(2-ethoxycarbonylphenyl)tris(p-tolyl)porphyrinato] zinc(II) (3). The zinc(II) porphyrin 3 has large affinity for pyrazole due to the hydrogen bond between pyrazole and the 2-ethoxycarbonyl group in addition to the coordination bonding accompanied by the conformational change of the ethoxycarbonyl group in the coordination process. The (1)H NMR, IR, and UV-vis spectra of 1 and its ESI-MS and VPO measurements have revealed the cyclic trimer structure with an overall association constant of 6.0 x 10(13) M(-2) at 22 degrees C. The contribution of the hydrogen bond to the total free energy change in trimer formation is estimated to be 7.5 kcal/mol based on a reference trimer system without a hydrogen bonding site. The trimer geometry causes characteristic exitonic interaction between porphyrin units to yield a broad Soret band which is deconvoluted into four components by UV-vis and MCD spectral analyses. Electrochemical measurements have shown that only the first ring-oxidation process proceeds stepwise in the trimer. PMID- 11421686 TI - Energy-transfer rate in crystals of double-complex salts composed of [Ru(N N)3](2+) (N-N = 2,2'-bipyridine or 1,10-phenanthroline) and [Cr(CN)6](3-): effect of relative orientation between donor and acceptor. AB - A block single-crystal was obtained using a diffusion method with a concentrated acetone-water (vol. 1/1) solution of [Ru(phen)(3)]Cl(2).6H(2)O (phen = 1,10 phenanthroline) and a concentrated aqueous solution of K(3)[Cr(CN)(6)], without evaporating solvents. The crystal was identified as a double-complex salt including two acetone and fourteen solvent water molecules, [Ru(phen)(3)](2)[Cr(CN)(6)]Cl.2(CH(3))(2)CO.14H(2)O (1). Measurement of the X-ray diffraction pattern of the double-complex salt was performed using an X-ray diffractometer with an Imaging-Plate (IP) Weissenberg camera. 1 crystallizes in the triclinic space group P1, with a = 13.930(5) A, b = 14.783(5) A, c = 11.137(6) A, alpha = 89.87(4) degrees, beta = 107.47(3) degrees, gamma = 96.68(3) degrees, and Z = 2. The crystal structure is very different from that of [Ru(bpy)(3)](2)[Cr(CN)(6)]Cl.8H(2)O (2) (bpy = 2,2'-bipyridine), which could be obtained using the same procedure and crystallizes in the monoclinic space group C2, with a = 22.414(2) A, b = 13.7686(15) A, c = 22.207(2) A, beta = 90.713(8) degrees, and Z = 4. The distance between the central-metal ions of ruthenium(II) and chromium(III) complexes in [Ru(phen)(3)](2)[Cr(CN)(6)]Cl.2(CH(3))(2)CO.14H(2)O (7.170 A) is shorter than that in [Ru(bpy)(3)](2)[Cr(CN)(6)]Cl.8H(2)O (9.173 A) by about 2 A, while the rate of energy transfer from the (3)MLCT state of [Ru(N-N)(3)](2+) to the (2)E(g) state of [Cr(CN)(6)](3-) in the former salt (9.5 x 10(5) s(-1)) is far slower than that in the latter one (6.0 x 10(6) s(-1)) at 77 K. These results indicate that the energy-transfer rate strongly depends, not upon the distance between central metal ions, rather, upon the mutual relative orientation between the donor and the acceptor complexes in double-complex salts. PMID- 11421687 TI - Crowded Cu(I) complexes involving benzo[h]quinoline: pi-stacking effects and long lives excited states. AB - The Friedlander condensation was employed to synthesize two series of 3,3' polymethylene bridged ligands, L, based on 2-(2'-pyridyl)-benzo[h]quinoline and 2,2'-bibenzo[h]quinoline (BHQ) along with the fully aromatic naphtho[1,2-b]-1,10 phenanthroline. Complexes [Cu(L)(2)](+) were prepared as their perchlorate or hexafluorophosphate salts. The solution state structures were analyzed by NMR and shielding effects reflected significant interligand pi-stacking interaction in the complexes. Solid-state structures of the complexes where L = 3,3' tetramethylene-2,2'-bibenzo[h]quinoline or naphtho[1,2-b]-1,10-phenanthroline were determined by X-ray analysis. The tetramethylene bridged complex showed a highly distorted coordination geometry with the BHQ rings of opposing ligands pi stacked at a interplanar distance of about 3.37 A. Complexes of the BHQ series showed a pronounced MLCT absorption maximum which shifted bathochromically from 496 to 610 nm as the 3,3'-bridge decreased from 4 to 2 carbons. The BHQ complexes luminesced strongly in CH(2)Cl(2) solution and the tetramethylene-bridged system showed the longest yet recorded excited-state lifetime for a copper MLCT excited state, tau = 5.3 micros and Phi = 0.10. PMID- 11421688 TI - Factors affecting the electronic ground state of low-spin iron(III) porphyrin complexes. AB - To determine the factors affecting the ground-state electron configuration of low spin Fe(III) porphyrin complexes, we have examined the (1)H NMR, (13)C NMR, and EPR spectra of a series of low-spin bis-ligated Fe(III) porphyrin complexes [Fe(Por)L(2)](+/-), in which the positions of porphyrin substituents and the coordination ability of axial ligands are different. The seven porphyrins used in this study are meso-tetraalkylporphyrins (TRP: R is propyl, cyclopropyl, or isopropyl), meso-tetraphenylporphyrin (TPP), meso-tetrakis(2,3,4,5,6 pentafluorophenyl)porphyrin, and 5,10,15,20-tetraphenyl-2,3,7,8,12,13,17,18 octaalkylporphyrins (ORTPP: R is methyl or ethyl). The porphyrin cores of TRP are more or less S(4)-ruffled depending on the bulkiness of the alkyl substituents, while those of ORTPP are highly S(4)-saddled. Three types of axial ligands are examined which have the following characteristics in ligand field theory: they are (i) strong sigma-donating imidazole (HIm), (ii) strong sigma-donating and weak pi-accepting cyanide (CN(-)), and (iii) weak sigma-donating and strong pi accepting tert-butyl isocyanide ((t)BuNC). In the case of the bis(HIm) complexes, only the isopropyl complex, [Fe(T(i)PrP)(HIm)(2)](+), has shown the less common (d(xz), d(yz))(4)(d(xy))(1) ground state; the other six complexes have exhibited the common (d(xy))(2)(d(xz), d(yz))(3) ground state. When the axial imidazole is replaced by cyanide, even the propyl and cyclopropyl complexes have shown the (d(xz), d(yz))(4)(d(xy))(1) ground state; the TPP and ORTPP complexes have still maintained the common (d(xy))(2)(d(xz), d(yz))(3) ground state. In the case of the bis((t)()BuNC) complexes, all the complexes have shown the (d(xz), d(yz))(4)(d(xy))(1) ground state. However, the contribution of the (d(xz), d(yz))(4)(d(xy))(1) state to the electronic ground state differs from complex to complex; the (d(xz), d(yz))(4)(d(xy))(1) contribution is the largest in [Fe(T(i)PrP)((t)()BuNC)(2)](+) and the smallest in [Fe(OETPPP)((t)BuNC)(2)](+). We have then examined the electronic ground state of low-spin [Fe(OEP)((t)BuNC)(2)](+) and [Fe(ProtoIXMe(2))((t)BuNC)(2)](+); OEP and ProtoIXMe(2) represent 2,3,7,8,12,13,17,18-octaethylporphyrin and protoporphyrin IX dimethyl ester, respectively. These porphyrins have a(1u) HOMO in contrast to the other seven porphyrins that have a(2u) HOMO. The (13)C NMR and EPR studies have revealed that the contribution of the (d(xz), d(yz))(4)(d(xy))(1) state in these complexes is as small as that in [Fe(OETPP)((t)BuNC)(2)](+). On the basis of these results, we have concluded that the low-spin iron(III) porphyrins that have (i) strong axial ligands, (ii) highly saddle shaped porphyrin rings, (iii) porphyrins with a(1u) HOMO, and (iv) electron withdrawing substituents at the meso positions tend to maintain the common (d(xy))(2)(d(xz), d(yz))(3) ground state. PMID- 11421689 TI - Syntheses, photophysics, and photochemistry of trinuclear copper(I) thiolate and hexanuclear copper(I) selenolate complexes: X-ray crystal structures of [Cu(6)(mu dppm)4(mu(3)-SePh)4](BF(4))2 and [Cu(6)(mu-(Ph(2)P)(2)NH(4))(mu(3) SePh)4](BF(4)2. AB - A series of trinuclear copper(I) thiolate complexes, [Cu(3)(mu-dppm)(3)(mu(3) SR)(2)]BF(4) (R = C(6)H(4)Cl-4, C(6)H(4)CH(3)-4, C(6)H(4)OCH(3)-4, C(6)H(4)(OCH(3))(2)-3,4, C(6)H(4)-benzo-15-crown-5, or (t)()Bu), [Cu(3)(mu dppm)(3)(mu(3)-S(t)()Bu)](BF(4))(2), and [Cu(3)(mu-dppm)(3)(mu(3)-SR)(mu(3) Cl)]BF(4) (R = C(6)H(4)CH(3)-4, C(6)H(4)(t)()Bu-4, or C(6)H(4)(CH(3))(3)-2,4,6) and two hexanuclear copper(I) selenolate complexes, [Cu(6)(mu-P(wedge)P)(4)(mu(3) SePh)(4)](BF(4))(2) (P(wedge)P = dppm, (Ph(2)P)(2)NH), have been synthesized, and their photophysical properties have been studied. The X-ray crystal structures of both copper(I) selenolate complexes have been determined. These complexes have been shown to exhibit long-lived low-energy emission in solution, attributed to an excited state of predominantly ligand-to-metal charge-transfer [chalcogenolate to copper(I)] origin. PMID- 11421690 TI - Zeolite-entrapped organized molecular assemblies. New evidence for highly efficient adjacent cage dyad formation and constrained rotational mobility of tris-ligated polypyridine complexes. AB - The primary objectives of the present work are to investigate the extent to which an entrapped, tris-ligated polypyridine complex of divalent ruthenium is restricted from rotating within the supercage of Y-zeolite and to provide additional evidence for the high efficiency of synthetic procedures developed for the preparation of adjacent cage dyads entrapped within the Y-zeolite framework. Specifically, the Y-zeolite entrapped complex, Z-[Ru(bpy)(2)(pypz)(2+) ] (where the prefix, Z, indicated a zeolite entrapped complex, bpy = 2,2'-bipyridine and pypz = 2-2-pyridylpyrazine), which contains only one reactive peripheral nitrogen atom, is shown to react with the reagent, (H(2)O)Ru(NH(3))(5)(2+), to yield the entrapped Z-[Ru(bpy)(2)(pypz)-Ru(NH(3))(5)](4+) complex. Treatment with a large excess of bpy, according to previously documented procedures, leads to the formation of the entrapped adjacent cage dyad, Z [Ru(bpy)(2)(pypz)(2+)/Ru(bpy)(3)(2+)], wherein the two-component complexes are entrapped in adjacent supercages. Spectroscopic measurements confirm the integrity of the component complexes and document a strong interaction between them. Most importantly, it is shown that a second treatment of this material with a large excess of the[(H(2)O)Ru(NH(3))(5)](2+) reagent does not lead to formation of significant amounts of Z-[Ru(bpy)(2)(pypz)-Ru(NH(3))(5)](4+); i.e., the presence of the Ru(bpy)(3)(2+) in the cage adjacent to the primary complex shields the latter from further reaction with the ruthenium pentammine reagent. This result demonstrates that, during the formation of the Ru(bpy)(3)(2+) secondary complex, the detached Ru(NH(3))(5)(2+) fragment does not drift into remote supercages, thereby providing unequivocal evidence for the high efficiency of the synthetic procedure for adjacent cage dyad formation. Furthermore, this result also makes it clear that rotation of the tris-ligated primary complex is restricted by the confinement of the Y-zeolite supercage to the extent that the single reactive peripheral nitrogen cannot be repositioned to a different window of the surrounding supercage. PMID- 11421691 TI - Ligand preorganization in metal ion complexation: molecular mechanics/dynamics, kinetics, and laser-excited luminescence studies of trivalent lanthanide complex formation with macrocyclic ligands TETA and DOTA. AB - The molecular mechanics and dynamics calculations, kinetics, and laser-excited luminescence studies were carried out for trivalent lanthanide (Ln(3+)) complexes of macrocyclic polyaminopolycarboxylate ligands TETA and DOTA (where TETA is 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid and DOTA is 1,4,7,10 tetraazacyclododecane-1,4,7,10-tetraacetic acid) to further understand the observed thermodynamic, kinetic, and structural properties and to examine how ligand preorganization affects metal ion complexation. Excitation spectroscopy (emission monitored at 614.0 nm) of the (7)F(0) --> (5)D(0) transition of Eu(3+) was used to study the aqueous properties of the Eu(3+)-TETA system. A stopped flow spectrophotometric method was used to study the formation kinetics of the aqueous Ce(3+)-TETA/DOTA systems in the pH range 6.1-6.7. Molecular mechanics calculation results are consistent with the proposed mechanism of Ln(DOTA)(-) formation, i.e., formation of a carboxylate O-bonded precursor, followed by metal ion moving into the preformed macrocyclic cavity. For Ln(TETA)(-) formation, at least two carboxylate O-bonded intermediates have been predicted and Ln(3+) ion assisted reorganization of the TETA ligand is present. The calculated bond distances and overall structures of Ln(DOTA)(-) and Ln(TETA)(-) were in agreement with the single-crystal and solution NMR structural data. The origin of the difference in thermodynamic stability of Ln(DOTA)(-) and Ln(TETA)(-) complexes and the corresponding formation intermediates is mainly due to the differences in water-occupancy energy (i.e., whether there is an apical coordinated water molecule), the ligand strain energy, and the cation-ligand interaction energy. Kinetic studies revealed that the formation rates of the Ce(TETA)(-) complex are smaller at lower pH and temperature but become greater at higher pH and temperature, as compared to those of the Ce(DOTA)(-) complex. This is attributed to the lanthanide ion and both mono- and di-hydroxide ion assisted TETA conformational reorganization and higher kinetic activation parameters. The presence of a di-hydroxide ion assisted intermediate rearrangement pathway could make the Ce(TETA)(-) complex formation rate faster at higher pH, and the higher activation barrier makes Ce(TETA)(-) complex formation rate slower at lower pH, as compared to those of the Ce(DOTA)(-) complex. PMID- 11421692 TI - Edge-bridged octahedral tungsten-oxygen-chlorine clusters: synthesis and characterization of two D(3d)-symmetric [W(6)O(6)Cl(12)](2-) isomers and [W(6)O(7)Cl(11)](3-). AB - Initial access to the chemistry of hexanuclear tungsten oxohalide clusters is provided through the reduction of WOCl(4) with bismuth metal at 360 degrees C. Reactions targeting W(6)O(6)Cl(10) produce an amorphous black solid, which, upon treatment with concentrated aqueous HCl, releases the edge-bridged octahedral cluster [alpha-W(6)O(6)Cl(12)](2-) into solution. The cluster exhibits a D(3d) symmetry structure in which the six oxygen atoms bridge the edges between two opposing triangular faces of a trigonally compressed W(6) octahedron. Reactions incorporating additional bismuth metal yield a mixture of soluble clusters, including a 5:7 ratio of [alpha-W(6)O(6)Cl(12)](2-) and another D(3d)-symmetry isomer, [beta-W(6)O(6)Cl(12)](2-). The latter species displays a different core structure, in which the six oxygen atoms are situated on the edges comprising two opposing triangular faces of a trigonally elongated W(6) octahedron. Isolated as the BuN(+) salts, the two isomers can be separated by a process relying on the differences in crystal morphology. Cyclic voltammetry of acetonitrile solutions shows two reversible one-electron reductions for each cluster, the alpha isomer being slightly more easily reduced. Density functional theory calculations indicate that the two isomers of [W(6)O(6)Cl(12)](2-) are nearly identical in energy, with the beta isomer lying just 1.4 kcal/mol below the alpha isomer. The other major product isolated from the reaction with additional bismuth is [W(6)O(7)Cl(11)](3-), a cluster at least formally related to [beta W(6)O(6)Cl(12)](2-) by substitution of an O(2)(-) ion for a core Cl(-) ion. In acetonitrile solution, this cluster displays a single reversible one-electron reduction. It is anticipated that the reactions elaborated here will lead to a general method for synthesizing metastable metal oxohalide clusters. PMID- 11421693 TI - Synthesis and characterization of ReCl(H(2))(AsMePh(2))(4), a classical hydride complex; reexamination of ReCl(H(2))(PMePh(2))(4) and theoretical calculations on model compounds. AB - The synthesis of ReCl(H(2))(AsMePh(2))(4) is reported and verified on the basis of (1)H NMR and FAB-MS. The minimum T(1) time for the metal-bonded hydrogen atoms was determined to be 44 ms (-78 degrees C) and 89 (-48 degrees C) ms at 200 and 400 MHz, respectively. From this minimum T(1) time, and, allowing for contributions from other nuclei to the relaxation, a metal-bonded H to H atom distance of 1.57 A can be assessed. The complex is reversibly oxidized at a potential E(1/2)(ox) = +0.07 V. The J(HD) coupling in the related ReCl(HD)(AsMePh(2))(4) compound was at 3.3 Hz. These data for ReCl(H(2))(AsMePh(2))(4) suggest that this molecule contains classical hydrides, and interpretations on T(1) and X-ray data published previously for ReCl(H(2))(PMePh(2))(4) suggest that this molecule contains a very long metal bonded H-H interaction of 1.39 A. Theoretical calculations on model compounds ReCl(H(2))(XR(3))(4) (X = P, As; R = H, Me) give virtually identical average d(H H) values of 1.5 (XH(3)) and 1.63 (XMe(3)) A. PMID- 11421694 TI - Syntheses, reactivity, and pi-donating ligand metathesis reaction of five coordinate sixteen-electron manganese(I) complexes: crystal structures of [Mn(CO(3)(-TeC(6)H(4)-o-NH-)](-), [(Mn(CO)(3))2(mu-SC(6)H(4)-o-S--S--C(6)H(4)-o mu-S--)], [(CO)(3)Mn(mu-SC(6)H(4)-o-NH(2)-)]2, and [(CO)(3)Mn(mu-SC(8)N(2)H(4)-o S-)](2)(2-). AB - The preparation of the varieties of five-coordinate sixteen-electron manganese(I) complexes [Mn(CO)(3)(-EC(6)H(4)-o-E'-)](-) (E = Te, Se, S, O; E' = NH, S, O) by (a) oxidative addition of 2-aminophenyl dichalcogenides to anionic manganese(0) carbonyl, (b) pi-donating ligand metathesis reaction of complex [Mn(CO)(3)( TeC(6)H(4)-o-NH-)](-), and (c) reduction /deprotonation of the neutral dimetallic [(Mn(CO)(3))(2)(mu-SC(6)H(4)-o-S-S-C(6)H(4)-o-mu-S-)]/[(CO)(3)Mn(mu-SC(6)H(4)-o NH(2)-)](2) proved successful approaches in this direction. The IR nu(CO) data of the coordinatively and electronically unsaturated [Mn(CO)(3)(-EC(6)H(4)-o-E'-)]( ) (E = Te, Se, S, O; E' = NH, S, O) complexes suggest the relative order of pi donating ability of the series of bidentate ligands being [TeC(6)H(4)-o-NH](2)(-) > [SeC(6)H(4)-o-NH](2)(-) > [SC(6)H(4)-o-NH](2)(-) > [SC(6)H(4)-o-S](2)(-) > [SC(6)H(4)-o-O](2)(-) > [OC(6)H(4)-o-O](2)(-). Proton NMR spectra of the [Mn(CO)(3)(-EC(6)H(4)-o-NH-)](-) (E = Te, Se, S) derivatives show the low-field shift of the amide proton ((1)H NMR (C(4)D(8)O): delta 9.66 (br) ppm (E = Te), 9.32 (br) ppm (E = Se), 8.98 (br) ppm (E = S)). The formation of the dimetallic [(CO)(3)Mn(mu-SC(8)N(2)H(4)-o-S-)](2)(2-) can be interpreted as coordinative association of two units of unstable mononuclear [(CO)(3)Mn(-SC(8)N(2)H(4)-o-S )](-) and reflects the pi-donating ability of the bidentate ligand is responsible for the formation of pentacoordinate, sixteen-electron manganese(I) carbonyl complexes. The neutral bimetallic manganese(I)-bismercaptophenyl disulfide complex [(Mn(CO)(3))(2)(mu-SC(6)H(4)-o-S-S-C(6)H(4)-o-mu-S-)] with internal S-S bond length of 2.222(1) A and the five-coordinate sixteen-electron complex [Mn(CO)(3)(-SC(6)H(4)-o-S-)](-) are chemically interconvertible. In a similar fashion, treatment of complex [Mn(CO)(3)(-SC(6)H(4)-o-NH-)](-) with HBF(4) yielded neutral dinuclear complex [(CO)(3)Mn(mu-SC(6)H(4)-o-NH(2)-)](2) and showed that the amine deprotonation is reversible. Investigations of pi-donating ligand metathesis reactions of complex [Mn(CO)(3)(-TeC(6)H(4)-o-NH-)](-) revealed that the stable intermediate, not the pi-donating ability of bidentate ligands, is responsible for the final protonation/oxidation product. This argument is demonstrated by reaction of [Mn(CO)(3)(-TeC(6)H(4)-o-NH-)](-) with 1,2 benzenedithiol, hydroxythiophenol, and catechol, respectively leading to the formation of [Mn(CO)(3)(-EC(6)H(4)-o-E'-)](-) (E = S, O; E' = S, O), although any pi-donor containing the amido group is a more effective donor than any other pi donor lacking an amido group. Also, the reactions of [Mn(CO)(3)(-TeC(6)H(4)-o-NH )](-) with electrophiles occurring at the more electron-rich amide site support that the more electron-rich amide donor of the chelating 2-tellurolatophenylamido occupies an equatorial site as indicated by a shorter Mn(I)-N bond length of the distorted trigonal bipyramidal [Mn(CO)(3)(-TeC(6)H(4)-o-NH-)](-). PMID- 11421695 TI - Hydrothermal synthesis and structural characterization of the (C(n)H(2n+6)N(2)[Mn(3)(HPO(3))4] (n = 3-8) new layered inorganic-organic hybrid manganese(II) phosphites. Crystal structure and spectroscopic and magnetic properties of (C(3)H(12)N(2)[Mn(3)(HPO(3))4]. AB - The (C(n)H(2n+6)N(2))[Mn(3)(HPO(3))(4)] (n = 3-8) compounds have been prepared by hydrothermal synthesis and characterized by X-ray diffraction data and spectroscopic techniques. The crystal structure of (C(3)H(12)N(2))[Mn(3)(HPO(3))(4)] has been solved from single-crystal X-ray diffraction. The unit-cell parameters are a = 9.502(1), b = 5.472(1), c = 14.523(4) A, beta = 95.01(3) degrees, monoclinic, C2/m, with Z = 2. The compound shows a layered structure stacked along the c-axis with the alkyldiammonium cations placed in the interlayer space. The sheets are formed by Mn(3)O(12) trimer units extended in the ab-plane and connected by (HPO(3))(2-) anions. The study of the (C(n)H(2n+6)N(2))[Mn(3)(HPO(3))(4)] (n = 4-8) phases by X-ray powder diffraction indicates an isotype relation with the propanediammonium compound. The Dq and Racah parameters calculated for (C(3)H(12)N(2))[Mn(3)(HPO(3))(4)] are Dq = 880, B = 660 and C = 3610 cm(-1). The ESR spectra show isotropic signals with a g-value of 2.008. Magnetic measurements indicate the presence of antiferromagnetic interactions inside the [Mn(3)(HPO(3))(4)](2-) sheets. The J/K value has been estimated to be -15 K by considering that the system behaves like an isolated trimer at high temperatures. PMID- 11421696 TI - Preparation and properties of gallaborane, GaBH(6): structure of the gaseous molecule H(2)Ga(mu-H)(2)BH(2) as determined by vibrational, electron diffraction, and ab initio studies, and structure of the crystalline solid at 110 K as determined by X-ray diffraction. AB - Gallaborane (GaBH(6), 1), synthesized by the metathesis of LiBH(4) with [H(2)GaCl](n) at ca. 250 K, has been characterized by chemical analysis and by its IR and (1)H and (11)B NMR spectra. The IR spectrum of the vapor at low pressure implies the presence of only one species, viz. H(2)Ga(mu-H)(2)BH(2), with a diborane-like structure conforming to C(2v) symmetry. The structure of this molecule has been determined by gas-phase electron diffraction (GED) measurements afforced by the results of ab initio molecular orbital calculations. Hence the principal distances (r(alpha) in A) and angles ( angle(alpha) in deg) are as follows: r(Ga.B), 2.197(3); r(Ga-H(t)), 1.555(6); r(Ga-H(b)), 1.800(6); r(B-H(t)), 1.189(7); r(B-H(b)), 1.286(7); angleH(b)-Ga-H(b), 71.6(4); and angleH(b)-B-H(b), 110.0(5) (t = terminal, b = bridging). Aggregation of the molecules occurs in the condensed phases. X-ray crystallographic studies of a single crystal at 110 K reveal a polymeric network with helical chains made up of alternating pseudotetrahedral GaH(4) and BH(4) units linked through single hydrogen bridges; the average Ga.B distance is now 2.473(7) A. The compound decomposes in the condensed phases at temperatures exceeding ca. 240 K with the formation of elemental Ga and H(2) and B(2)H(6). The reactions with NH(3), Me(3)N, and Me(3)P are also described. PMID- 11421697 TI - Photophysics of pyridylporphyrin Ru(II) adducts: heavy-atom effects and intramolecular decay pathways. AB - Eight adducts between different pyridylporphyrins and ruthenium complexes, MPyP[RuCl(2)(DMSO)(2)(CO)], c-DPyP[RuCl(2)(DMSO)(2)(CO)](2), TrPyP[RuCl(2)(DMSO)(2)(CO)](3), TPyP[RuCl(2)(DMSO)(2)(CO)](4), (MPyP)(2)[RuCl(2)(DMSO)(2)], [c-DPyP[RuCl(2)(DMSO)(2)]](2), MPyP[RuCl(2)(CO)(3)], and [c-DPyP[RuCl(2)(CO)(2)]](2), have been investigated. The results show that in all the adducts the porphyrin singlet is quenched, to a greater or lesser extent, relative to the parent-free molecule. This study provides insight into the mechanisms of singlet quenching in the adducts. Two mechanisms for singlet quenching, both related to the "heavy-atom effect" of the ruthenium center and experimentally distinguishable by transient spectroscopy, are examined. Enhanced intersystem crossing within the porphyrin chromophore is demonstrated for the series of adducts MPyP[RuCl(2)(DMSO)(2)(CO)], c-DPyP[RuCl(2)(DMSO)(2)(CO)](2), TrPyP[RuCl(2)(DMSO)(2)(CO)](3), and TPyP[RuCl(2)(DMSO)(2)(CO)](4), where a nice correlation is observed between the magnitude of the effect and the number of ruthenium centers attached to the pyridylporphyrin chromophore. Singlet-triplet energy transfer from the pyridylporphyrin chromophore to the ruthenium center(s) is an additional efficient quenching channel for adducts containing ruthenium centers with weak field ligands and low triplet energies, such as (MPyP)(2)[RuCl(2)(DMSO)(2)] and [c-DPyP[RuCl(2)(DMSO)(2)]](2). PMID- 11421698 TI - Metal-capped alpha-cyclodextrins: squaring the circle. AB - This report describes the four-step synthesis of 6A,6D-diamino-6A,6D-dideoxy 2(A),2(B),2(C),2(D),2(E),2(F),3(A),3(B),3(C),3(D),3(E),3(F), 6(B),6(C),6(E),6(F) hexadeca-O-methylcyclomaltohexaose (5), a methylated alpha-CD that bears two CH(2)NH(2) ligands located on diametrically opposed glucopyranose rings. Reaction of 5 with K(2)PtCl(4) afforded the water-soluble chelate complex [PtCl(2)(5)] where the metal center is bonded to cis-arranged nitrogen atoms. A single-crystal X-ray structure of the latter reveals the high distortion imposed on the CD structure by the short metallo-organic cap. The cyclodextrin core adopts an unprecedented elongated, almost rectangular shape, the shortest and longest O(4)(n). O(4)(n+3) distances being respectively 5.44 and 9.98 A. Two opposing glucose rings are no longer in the usual (4)C(1) chair conformation, but adopt an elongated (O)S(2) skew-boat structure. The observed CD-flattening produces a highly preorganized hydrophilic pocket that complexes through multiple hydrogen bonding a single water molecule. Complex [PtCl(2)(5)] and the corresponding synthetic intermediates were characterized by elemental analysis, MS, and IR and NMR spectroscopy. PMID- 11421699 TI - On the electronic coupling of the C=O stretching vibrations in Cl(2)Pt(malonate)( ). AB - Ab initio calculations on the complex Cl(2)Pt(malonate)(-) have been performed to estimate the magnitude and the source of the splitting between the symmetric and antisymmetric C=O stretches. It is shown that a large part of the splitting can result from pure electronic coupling mediated via the pi-structure of the ligand. Calculations have also been performed on the free ligand malonate(-) and the neutral malonaldehyde that show decreasing electronic coupling, which is consistent with the decreasing resonance stabilization. A simple effective C=O interaction potential has been produced for Cl(2)Pt(malonate)(-) that is used to estimate the electronic splitting of the 1600 cm(-1) IR band to 137 cm(-1). Anharmonic contributions seem to reduce this value somewhat, but the magnitude and sign of the splitting is similar to the experimental value of 183 cm(-1) found for the similar Cl(2)Pt(acac)(-) complex. PMID- 11421700 TI - Solvent effects on uranium(VI) fluoride and hydroxide complexes studied by EXAFS and quantum chemistry. AB - The structures of the complexes UO(2)F(n)(H(2)O)(5-n)(2-n), n = 3-5, have been studied by EXAFS. All have pentagonal bipyramid geometry with U-F of and U-H(2)O distances equal to 2.26 and 2.48 A, respectively. On the other hand the complex UO(2)(OH)(4)(2-) has a square bipyramid geometry both in the solid state and in solution. The structures of hydroxide and fluoride complexes have also been investigated with wave function based and DFT methods in order to explore the possible reasons for the observed structural differences. These studies include models that describe the solvent by using a discrete second coordination sphere, a model with a spherical, or shape-adapted cavity in a conductor-like polarizable continuum medium (CPCM), or a combination of the two. Solvent effects were shown to give the main contribution to the observed structure variations between the uranium(VI) tetrahydroxide and the tetrafluoride complexes. Without a solvent model both UO(2)(OH)(4)(H(2)O)(2-) and UO(2)F(4)(H(2)O)(2-) have the same square bipyramid geometry, with the water molecule located at a distance of more than 4 A from uranium and with a charge distribution that is very near identical in the two complexes. Of the models tested, only the CPCM ones are able to describe the experimentally observed square and pentagonal bipyramid geometry in the tetrahydroxide and tetrafluoride complexes. The geometry and the relative energy of different isomers of UO(2)F(3)(H(2)O)(2-) are very similar, indicating that they are present in comparable amounts in solution. All calculated bond distances are in good agreement with the experimental observations, provided that a proper model of the solvent is used. PMID- 11421701 TI - Hybrid molecular materials based upon organic pi-electron donors and metal complexes. Radical salts of bis(ethylenethia)tetrathiafulvalene (BET-TTF) with the octahedral anions hexacyanoferrate(III) and nitroprusside. The first kappa phase in the BET-TTF family. AB - The synthesis, structure, and physical characterization of two new radical salts formed with the organic donor bis(ethylenethia)tetrathiafulvalene (BET-TTF) and the octahedral anions hexacyanoferrate(III), [Fe(CN)(6)](3-), and nitroprusside, [Fe(CN)(5)NO](2-), are reported. These salts are (BET TTF)(4)(NEt(4))(2)[Fe(CN)(6)] (1) (monoclinic space group C2/c with a = 38.867(7) A, b = 8.438(8) A, c = 11.239(6) A, beta = 90.994(9) degrees, V = 3685(4) A(3), Z = 4) and (BET-TTF)(2)[Fe(CN)(5)NO].CH(2)Cl(2) (2) (monoclinic space group C2/c with a = 16.237(6) A, b = 18.097(8) A, c = 12.663(7) A, beta = 106.016(9) degrees, V = 3576(3) A(3), Z = 4). In salt 1 the organic BET-TTF molecules are packed in orthogonal dimers, forming the first kappa phase observed for this donor. The analysis of the bond distances and the electronic and IR spectra suggests a degree of ionicity of 1/4 per BET-TTF molecule, in agreement with the stoichiometry of the salt. The electrical properties show that 1 is a semiconductor with a high room-temperature conductivity (11.6 S cm(-1)) and a low activation energy (45 meV), in agreement with the band structure calculations. The magnetic susceptibility of 1 shows, besides the paramagnetic contribution from the anion, a temperature-independent paramagnetism (TIP) of the Pauli type due to the electronic delocalization observed at high temperatures in the organic sublattice. This Pauli type paramagnetism is confirmed by the ESR spectra that also show a Dysonian line when the magnetic field is parallel to the conducting plane, typical of metallic and highly conducting systems. Salt 2 presents an unprecedented packing of the organic molecules that form zigzag tunnels where the anions and the solvent molecules are located. The stoichiometry indicates that all the BET-TTF molecules bear a charge of +1, and accordingly, 2 behaves as a semiconductor with a very low room-temperature conductivity. The magnetic properties of this salt indicate that the unpaired electrons on the organic molecules are strongly antiferromagnetically coupled, giving rise to a diamagnetic behavior of 2, as the nitroprusside anion is also diamagnetic. PMID- 11421702 TI - "Intermolecular" trapping of a nonheme Fe(IV)=O intermediate. AB - The reactions of Fe(III)(TPA) (TPA = tris(2-pyridylmethyl)amine) complexes with 2 methyl-1-phenyl-2-propyl-hydroperoxide (MPPH) delivered by syringe pump under anaerobic conditions afford nearly quantitative conversion of MPPH to products derived from the benzyl radical. These results unequivocally show that MPPH breaks down by O-O bond homolysis, leading to the formation of the benzyl radical and a high valent Fe(IV)=O species. Without added substrates, the benzyl radical reacts with the high valent species to form benzyl alcohol or benzyl halides. The Fe(IV)=O species can also effect the two-electron oxidation of added substrates such as thioanisole, cyclohexanol, and cyclooctene under appropriate conditions. The oxidation of thioanisole and cyclohexanol is likely facilitated by pre equilibrium binding of the substrate to the metal center, allowing these substrates to intercept the high valent iron-oxo species as it forms. These results suggest the importance of close proximity to direct the high valent metal center down a desired pathway. PMID- 11421703 TI - Preparation, crystal structure, and spectroscopic characterization of [(Se(2)SN(2)Cl](2). AB - The reaction of [(Me(3)Si)(2)N](2)S with an equimolar amount of SeCl(4) in dioxane at 50 degrees C affords [(Se(2)SN(2))Cl](2) (1) in excellent yield. Crystals of 1 are orthorhombic, space group Pbca, with a = 8.5721(7) A, b = 7.8336(6) A, c = 15.228(1) A, and Z = 8. The crystal structure contains two planar Se(2)SN(2)(*)(+) rings which are linked by intermolecular Se.Se interactions [d(Se-Se) = 3.0690(7) A]. The EI mass spectrum shows Se(2)SN(2)(*)(+) as the fragment of highest mass. Both the (14)N and (77)Se NMR spectra show a single resonance (-52 and 1394 ppm, respectively). The solid [(Se(2)SN(2))Cl](2) gives a strong ESR signal indicating the presence of a Se(2)SN(2)(*)(+) radical. The Raman spectrum was assigned through normal coordinate treatment involving a general valence force field. The vibrational analysis yielded a good agreement between the observed and calculated wavenumbers. PMID- 11421704 TI - Syntheses, structures, and physicochemical properties of diruthenium compounds of tetrachlorocatecholate with metal-metal bonded Ru(3+)(mu-OR)(2)Ru(3+) and Ru(3.5+)(mu-OR)(2)Ru(3.5+) cores (R = CH(3) and C(2)H(5)). AB - Metal-metal bonded Ru(3+)(mu-OR)(2)Ru(3+) and Ru(3.5+)(mu-OR)(2)Ru(3.5+) (R = CH(3) and CH(3)CH(2)) compounds with tetrachlorocatecholate (Cl(4)Cat) have been synthesized in the corresponding alcohol, MeOH and EtOH, from a nonbridged Ru(2+) Ru(3+) compound, Na(3)[Ru(2)(Cl(4)Cat)(4)(THF)].3H(2)O.7THF (1). In alcohol solvents, compound 1 is continuously oxidized by oxygen to form Ru(3+)(mu OR)(2)Ru(3+) and Ru(3.5+)(mu-OR)(2)Ru(3.5+) species. The presence of a characteristic countercation leads to selective isolation of either Ru(3+)(mu OR)(2)Ru(3+) or Ru(3.5+)(mu-OR)(2)Ru(3.5+) as a stable adduct species. In methanol, Ph(4)PCl and dibenzo-18-crown-6-ether afford Ru(3+)(mu-OMe)(2)Ru(3+) species, [A](2)[Ru(2)(Cl(4)Cat)(4)(mu-OMe)(2)Na(2)(MeOH)(6)] ([A](+) = Ph(4)P(+) (2), [Na(dibenzo-18-crown-6)(H(2)O)(MeOH)](+) (3)), while benzo-15-crown-5-ether provides a Ru(3.5+)(mu-OMe)(2)Ru(3.5+) species, [Na(benzo-15-crown 5)(2)][Ru(2)(Cl(4)Cat)(4)(mu-OMe)(2)Na(2)(MeOH)(6)] (4). The air oxidation of 1 in a MeOH/EtOH mixed solvent (1:1 v/v) containing benzo-15-crown-5-ether provides a Ru(3.5+)(mu-OMe)(2)Ru(3.5+) species, [Na(benzo-15-crown 5)(H(2)O)][Ru(2)(Cl(4)Cat)(2)(mu-OMe)(2)Na(2)(EtOH)(2)(H(2)O)(2)(MeOH)(2)].(benzo 15-crown-5) (5). Similarly, the oxidation of 1 in ethanol with Ph(4)PCl provides a Ru(3.5+)(mu-OEt)(2)Ru(3.5+) species, (Ph(4)P)[Ru(2)(Cl(4)Cat)(4)(mu OEt)(2)Na(2)(EtOH)(6)] (7). A selective formation of a Ru(3+)(mu-OEt)(2)Ru(3+) species, (Ph(4)P)(2)[Ru(2)(Cl(4)Cat)(4)(mu-OEt)(2)Na(2)(EtOH)(2)(H(2)O)(2)] (6), is found in the presence of pyrazine or 2,5-dimethylpyrazine. The crystal structures of these compounds, except 2 and 7, have been determined by X-ray crystallography, and all compounds have been characterized by several spectroscopic and magnetic investigations. The longer Ru-Ru bonds are found in the Ru(3+)(mu-OR)(2)Ru(3+) species (2.606(1) and 2.628(2) A for 3 and 6, respectively) compared with those of Ru(3.5+)(mu-OMe)(2)Ru(3.5+) species (2.5260(6) A and 2.514(2) A for 4 and 5, respectively). These structural features and magnetic and ESR data revealed the electronic configurations of sigma(2)pi(2)delta(2)delta(2)pi(2) and sigma(2)pi(2)delta(2)delta(2)pi(1) for Ru(3+)(mu-OR)(2)Ru(3+) and Ru(3.5+)(mu-OR)(2)Ru(3.5+), respectively, in which the former is diamagnetic and the latter is paramagnetic with S = (1)/(2) ground state. Compound 5 forms a one-dimensional chain with alternating arrangement of a Ru(3.5+)(mu-OMe)(2)Ru(3.5+) unit and a free benzo-15-crown-5-ether molecule by intermolecular hydrogen bonds (O(H(2)O).O(crown-ether) = 2.91-3.04 A). The cyclic voltammetry in DMF affords characteristic metal-origin voltammograms; two reversible and two quasi-reversible redox waves were observed. The feature of cyclic voltammograms for the Ru(3+)(mu-OR)(2)Ru(3+) species (2, 3, and 6) and the Ru(3.5+)(mu-OR)(2)Ru(3.5+) species (4 and 7) are similar to each other, indicating that both species are electrochemically stable. The isolation of the pyrazine-trans-coordinated species, [Ph(4)P][Ru(Cl(4)Cat)(2)(L)(2)] (L = pyrazine (8), 2,5-dimethylpyrazine (9)), revealed the selective isolation of 6 from pyrazine-containing solution. UV-vis spectral variation by ethanolysis for 9 demonstrated the selective conversion from the pyrazine-trans-coordinated species to the Ru(3+)(mu-OEt)(2)Ru(3+) species without an oxidation to the Ru(3.5+)(mu OEt)(2)Ru(3.5+) species. This result suggests the presence of equilibrium between [Ru(Cl(4)Cat)(2)(L)(2)](-) and Ru(3+)(mu-OEt)(2)Ru(3+) species in the synthetic condition for 6. PMID- 11421705 TI - Technetium(V) oxo complexes of substituted propylene diamine dioxime (PnAO) ligands: water-dependent interconversion between syn and anti isomers. AB - 99mTc and (99)Tc complexes of PnAO (propylene diamine dioxime) ligands monosubstituted in the 6-position [PnAO-6-R] were prepared and studied. Ligands substituted with an alkyl group or with no substituent (R = H, CH(3), or CH(2)CH(CH(3))(2)), gave only one Tc complex. However, for several other nonalkyl substituents (R = COOCH(3), OH, OCH(3), OCH(2)CH(3), F, CN, NHCOCH(3), and NHCOCH(2)CH(3)), two Tc complexes A and B were formed. Products A and B were assigned to the anti and syn TcO(PnAO-6-R) species, respectively, based on (1)H NMR results. X-ray structure analyses supported these assignments. The A (anti) isomer of TcO(PnAO-6-OH) had the chemical formula TcC(13)H(25)N(4)O(4) and crystallized in an orthorhombic system with space group P2(1)2(1)2(1) and Z = 4; a = 12.744(2) A, b = 13.591(2) A, c = 9.976(2) A. The B (syn) isomer of TcO(PnAO 6-CN) had the chemical formula TcC(14)H(24)N(5)O(3) and was a 1:4 mixture of two monoclinic polymorphs: individual rectangular prisms (space group P2(1)/c, Z = 4) and clusters of intergrown twinned rectangular rods (space group Cc, Z = 8). For the prisms, a = 12.457(1) A, b = 13.932(1) A, c = 10.336(1) A, and for the rods, a = 31.344(5) A, b = 6.993(1) A, c = 21.657(2) A. The syn and anti isomers interconverted in the presence of water; nonequilibrium mixtures of epimers remained unchanged under dry conditions. The HPLC behavior under reversed phase conditions was consistent with on-column interconversion (poor resolution), whereas the two isomers were cleanly resolved under drier normal phase conditions. An oxo inversion mechanism involving trans water attack is proposed for the interconversion process. Water also influenced the position of equilibrium of the two isomers. The syn isomer was stabilized in water relative to the anti isomer. PMID- 11421706 TI - Toward designed assembly of microporous coordination networks constructed from silver(I)-hexamethylenetetramine layers. AB - Eight interesting microporous networks based on 2-D [Ag(mu(3)-hmt)] (hmt = hexamethylenetetramine) layers were obtained via rational synthetic strategies. Out of these products isolated and structurally characterized, six present a metal-to-hmt molar ratio of 1:1 and contain 2-D coordination layers assembled by 2-D Ag(I)-hmt networks with aromatic monocarboxylates, which are [Ag(mu(3)-hmt)(p nba)].2.5H(2)O (1), [Ag(mu(3)-hmt)(m-nba)].2.5H(2)O (2), [Ag(mu(3)-hmt)(dnba)] (3), [Ag(mu(3)-hmt)(alpha-hna)](EtOH) (4), [Ag(mu(3)-hmt)(beta-hna)](EtOH) (5), and [Ag(mu(3)-hmt)(noa)](H(2)O)(EtOH) (6) (p-nba = 4-nitrobenzoate, m-nba = 3 nitrobenzoate, dnba = 3,5-dinitrobenzoate, alpha-hna = 1-hydroxy-2-naphthate, beta-hna = 3-hydroxy-2-naphthate, and noa = 2-naphthoxyacetate), and two present a metal-to-hmt molar ratio of 2:1 and contain 3-D microporous networks constructed from the 2-D Ag-hmt layers and linear dicarboxylates as molecular pillars, which are [Ag(2)(mu(3)-hmt)(2)(fa)].4H(2)O (7) and [Ag(2)(mu(3) hmt)(2)(adp)].8H(2)O (8) (fa = fumarate and adp = adipate). The result shows that the pore sizes may be tuned with different carboxylates. Gas adsorption measurements were performed to confirm the microporosity of these coordination open frameworks. PMID- 11421707 TI - [N(2)H(5)](+)(2)[N(4)C-N=N-CN(4)](2-): a new high-nitrogen high-energetic material. AB - The reaction of [N(2)H(5)](+)(2)[SO(4)](2-) with barium 5,5'-azotetrazolate gave new high-energy-density materials (HEDM) based on the 5,5'-azotetrazolate dianion. The dihydrazinium salt of [N(4)C-N=N-CN(4)](2-) 1, its dihydrate 2, and its dihydrazinate 3 were prepared in high yield. Synthesis in water afforded yellow needles of [N(2)H(5)](2)(+)[N(4)C-N=N-CN(4)](2-).2H(2)O (2): monoclinic, P2/c, a = 8.958(2) A, b = 3.6596(7) A, c = 16.200(3) A, beta = 96.834(3) degrees, V = 527.3(2) A(3), Z = 2; synthesis in anhydrous hydrazine gave yellow [N(2)H(5)](2)(+)[N(4)C-N=N-CN(4)](2-).2N(2)H(4) (3): triclinic, P1, a = 4.6208(6) A, b = 8.585(1) A, c = 9.271(1) A, alpha = 108.486(2) degrees, beta = 95.290(2) degrees, gamma = 102.991(2) degrees, V = 334.51(8) A(3), Z = 1. The compounds were characterized by elemental analysis and vibrational (IR, Raman) and multinuclear NMR spectroscopy ((1)H, (13)C, (14)N, (15)N). The new compounds represent new high-nitrogen HEDMs with one of the highest nitrogen contents reported to date ([N(2)H(5)](+)(2)[N(4)C-N=N-CN(4)](2-) 85.2%; [N(2)H(5)](+)(2)[N(4)C-N=N-CN(4)](2-).2H(2)O 73.3%; [N(2)H(5)](+)(2)[N(4)C-N=N CN(4)](2-).2N(2)H(4) 85.7%). The standard heat of formation of the solvate-free compound 1 was computed at the MP2(FULL)/6-311+G(d,p) level of theory to be DeltaH degrees (f) = 264 kcal mol(-1), which translates to 1147 kcal kg(-1) and is one of the highest ever reported. The compounds are stable at room temperature, almost insensitive to friction and impact, but detonate violently when the explosion is initiated, e.g., by rapid heating over the decomposition temperature or by using an initiator. PMID- 11421708 TI - Correlation of structure and triboluminescence for tetrahedral manganese(II) compounds. AB - To investigate the structural basis of triboluminescence, several known tetrahedrally coordinated Mn(II) complexes have been synthesized according to literature methods and their crystal structures have been determined by X-ray diffraction. Among them, (MePh(3)P)(2)[MnCl(4)] (2), a = 15.4804(4) A, cubic, space group P2(1)3, Z = 4; (Et(4)N)(2)[MnBr(4)] (4), a = 13.362(1) A, c = 14.411(1) A, tetragonal, space group P42(1)m, Z = 4; MnBr(2)(OPPh(3))(2) (7), a = 9.974(1) A, b = 10.191(3) A, c = 10.538(2) A, alpha = 65.32(1) degrees, beta = 63.49(1) degrees, gamma = 89.44(2) degrees, triclinic, space group P1, Z = 1; and MnBr(2)(OAsPh(3))(2) (10), a = 17.816(3) A, b = 10.164(1) A, c = 18.807(3) A, orthorhombic, space group Pca2(1), Z = 4 were reported to be triboluminescent and (Me(4)N)(2)[MnCl(4)] (3), a = 9.016(3) A, b = 36.90(2) A, c = 15.495(3) A, beta = 90.72(3) degrees, monoclinic, space group P2(1)/n, Z = 12, and MnI(2)(OAsPh(3))(2) (11), a = 10.094(4) A, b = 10.439(2) A, c = 34.852(2) A, alpha = 83.17(4) degrees, beta = 86.09(2) degrees, gamma = 75.16(3) degrees, triclinic, space group P1, Z = 4, were reported to be not triboluminescent. The result supports the correlation between space group acentricity and triboluminescence activity. PMID- 11421709 TI - Electrochemical assembling/disassembling of helicates with hysteresis. AB - A series of eight tetradentate, ditopic, bisimino bisheterocyclic ligands (1-8), and their complexes with Cu(I) and Cu(II), have been studied in CH(3)CN solution, by means of (1)H NMR, mass, and UV/vis spectroscopy, while the crystal and molecular structure of the Cu(II) complexes [Cu(3)](CF(3)SO(3))(2) and [Cu(4)](CF(3)SO(3))(2) and of the Cu(I) complexes [Cu(2)(4)(2)](ClO(4))(2) and [Cu(2)(5)(2)](ClO(4))(2) have been determined by X-ray diffraction methods. The Cu(II) complexes are monomeric, almost square-planar structures, both in solution and in the solid state, while the Cu(I) complexes are two-metal, two-ligand dimers which can be both helical and "box-like" in the solid, while they adopt a simple helical configuration in acetonitrile solution. The systems made of ligands 1-8 and copper are bistable, as under the same conditions either the Cu(I) helical dimers or the Cu(II) monomers can be obtained and are stable. The electrochemical behavior of the 16 copper complexes has been studied in acetonitrile solutions by cyclic voltammetry. One reduction and one oxidation wave were found in all cases, which display no return wave and are separated by a 500-1000 mV interval. Irreversibility is due to the fast self-assembling process that follows the reduction of [Cu(II)(L)](2+) and to the fast disassembling process that follows the oxidation of [Cu(I)(2)(L)(2)](2+) (L = 1-8). However, the overall [oxidation+disassembling] or [reduction+self-assembling] processes, i.e., [Cu(I)(2)(L)(2)](2+) = 2[Cu(II)(L)](2+) + 2e(-), are fully reversible. Moreover, CV profiles show that solutions containing copper and L undergo hysteresis on changing the applied electrochemical potential: in the same potential interval, the systems can exist in solution as either [Cu(I)(2)(L)(2)](2+) or [Cu(II)(L)](2+), depending on the electrochemical history of the solution. Moreover, by changing the structural or donor features of the ligands it is possible to modulate the potentials at which the system undergoes a transition from one to the other of its two possible states, in the hysteresis cycle. In addition, the spectral properties of the Cu(I) and Cu(II) complexes of the considered ligands make these systems good candidates for storing information in solution, which can be electrochemically written or erased and spectroscopically read. PMID- 11421710 TI - La(3+)-catalyzed methanolysis of phosphate diesters. Remarkable rate acceleration of the methanolysis of diphenyl phosphate, methyl p-nitrophenyl phosphate, and bis(p-nitrophenyl) phosphate. AB - The interaction of La(3+) with diphenyl phosphate, methyl p-nitrophenyl phosphate, and bis(p-nitrophenyl) phosphate (4, 5, and 3) and the ensuing catalysis of methanolysis has been studied in methanol under completely homogeneous conditions of known pH. (31)P NMR shows that phosphates 3 and 4 are each associated with La(3+) as a rapidly interconverting mixture of complexes having 2:1, 1:1 (or 2:2), and 1:2 stoichiometry. At [La(3+)] > 2 x 10(-4) M, the kinetically dominant species in the pH regions of interest are phosphate-bound La(3+) dimers having 2-5 associated methoxides. Potentiometric titration reveals that 4 mM La(3+) alone in MeOH exhibits two apparent pK(a)s at 7.86 and 10.44 consuming 1 and 1.5 CH(3)O(-) per metal, the suggested structures being La(3+) dimers with first two and then five associated methoxides. In the presence of 0.5 equiv of 4 the potentiometric titration reveals strong complexation of 4 to La(3+) and a profile with apparent pK(a)s at 7.8 and 11.1 consuming 1 and 1.5 CH(3)O(-) per metal corresponding to phosphate-bridged La(3+) dimers with first two and then five associated methoxides. Methanolysis of 3-5 is strongly accelerated by La(3+). At pH 8.9 or 11.1, respectively, as little as 5 x 10(-4) M La(OTf)(3) accelerates the methanolysis of 3 or 4 by 10(10)-fold relative to the background reaction. Detailed kinetic studies of the methanolysis of 5 at varying [La(3+)] and pH indicate that both La(3+) monomers and dimers with associated methoxides are reactive species. The rate constants for La(3+) monomer and dimer catalysis of the methanolysis of 5 are dependent on pH, the slopes of the log plots being 0.35 and 0.5, respectively. A scheme is proposed to account for the non-first-order dependence on [CH(3)O(-)] wherein the La(3+)(CH(3)O(-))(x) + 5 <- > La(3+)(CH(3)O(-))(x):5 and (La(3+))(2)(CH(3)O(-))(y) + 5 <--> 5:(La(3+))(2)(CH(3)O(-))(y) equilibria are driven to the left with increasing numbers of associated methoxides and that attack on these complexes, whether by external or metal-associated methoxide, is slower due to the reduced net positive charge on the complex. PMID- 11421711 TI - K(2)SrIn(7): an electron-deficient indium network structure that reflects limitations of cation accommodation. synthesis, structure, and bonding. AB - A new phase in the K-Sr-In system was discovered following direct fusion of the neat elements in a niobium tube at 900 degrees C and equilibration at 700 degrees C for 5 days. Single-crystal X-ray diffraction analysis reveals that K(2)SrIn(7) crystallizes in an orthorhombic system, space group Cmcm, Z = 4, a = 5.0455(5) A, b = 11.960(2) A, c = 19.762(4) A. The structure contains a three-dimensional In network built of sheets of condensed pentagonal prismatic columns interbonded along. Two rather different types of channels are separately occupied by K and Sr atoms, the latter centered in 15-atom indium polyhedra. Band structure calculations and resistivity and susceptibility measurements indicate that the compound is metallic and diamagnetic. The one-electron deficiency in the valence band per formula unit brought on by the limited cation count is analyzed in terms of the character of the bonding, some of that from multicenter In bonding falling above E(F). PMID- 11421712 TI - Subtle bite-angle influences on N(2)S(2)Ni complexes. AB - A new N(2)S(2)Ni complex based on the 1,4-diazacycloheptane (dach) framework allows the study of the effects of ring size, in fused diamines, on the structural and chemical properties of nickel(II) dithiolate and dithioether complexes. Compared to its 1,5-diazacyclooctane (daco) derivatives, the dithiolate complex (bmedach)Ni and the S-templated, macrocyclic dithioether complex (propyl-bmedach)NiBr(2) show decreased cavity sizes with narrower angleN Ni-N angles (by ca. 6 degrees ) and wider angleS-Ni-S angles (also by ca. 6 degrees ). The electrochemical properties of the dithiolate complexes based on dach and daco are nearly identical, while the (propyl-bmedach)NiBr(2) complex shows a 140 mV destabilization of the Ni(I) oxidation state relative to its daco analogue. Molecular structures for the (bmedach)Ni and (propyl-bmedach)NiBr(2) complexes and their respective electrochemical and spectroscopic properties are reported. PMID- 11421714 TI - Is borazine aromatic? Unusual parallel behavior between hydrocarbons and corresponding B-N analogues. PMID- 11421713 TI - Cross-electron-transfer reactions of the [Cu(II/I)(bite)](2+/+) redox couple. AB - A series of outer-sphere cross-electron-transfer reactions involving the [Cu(bite)](2+/+) redox couple has been investigated in acetonitrile at 25 degrees C. In this complex, the bite ligand is a macrocyclic N(2)S(2) ligand with a 2,2' biphenyl moiety as its backbone. The reaction of [Cu(II)(bite)](2+) with [Ru(hfac)(3)](-) produces [Cu(I)(bite)](+) and [Ru(hfac)(3)] reversibly with K = 1.9. The rate law is second order in both directions, with a rate constant of (8.22 +/- 0.27) x 10(2) M(-1) s(-1) in the forward direction. Rate constants were also obtained for the irreversible reactions of three Co(II) clathrochelates with [Cu(bite)](2+). The oxidation of [Cu(bite)](+) by [Fe(bpy)(3)](3+) was studied in order to obtain a rate constant for oxidation as well as reduction. Application of the Marcus cross relationship to these rate constants gives apparent self exchange rate constants that are reasonably consistent yet unusually low, with an average value of 1.0 x 10(-2) M(-1) s(-1). The self-consistence of the apparent self-exchange rate constants implies that all of the cross reactions proceed through the same intermediate, and hence, the outer-sphere self-exchange reaction should have a second-order rate law with k = 1.0 x 10(-2) M(-1) s(-1). The much faster first-order self-exchange process reported previously for the [Cu(bite)](2+/+) couple in acetone implies a more efficient mechanism for the self-exchange reaction than for the cross reactions, such as an inner-sphere mechanism. Cyclic voltammograms of [Cu(bite)](n+) are strongly sensitive to the nature of the working electrode, thus precluding the use of these data in interpreting the homogeneous redox kinetics. PMID- 11421715 TI - Ferromagnetic coupling in a one-dimensional molecular railroad copper(II) azido compound containing a defective double cubane motif. PMID- 11421716 TI - Synthesis, crystal structure, and magnetic properties of the mixed-ligand complex [Gd(CF(3)CO(2)(3)(phen)(2)(H2O)]. PMID- 11421717 TI - Natural abundance (15)N CP MAS NMR as a novel tool for investigating metal binding to nucleotides in the solid state. PMID- 11421718 TI - Variable coordination modes in dialkyldithiophosphinato complexes of group 13 metals: the X-ray single crystal structures of tris(diisobutyldithiophosphinato)gallium(III) and -indium(III). PMID- 11421719 TI - Iron(II) complexes containing a ferrocenyl framework attached to 2,2'-bipyridine or 1,10-phenanthroline subunits: formation of stable Fe-bis(2,2'-bipyridine)-like and Fe-bis(1,10-phenanthroline)-like complexes. PMID- 11421720 TI - Improved syntheses of one-dimensional cyanide-bridged lanthanide-transition-metal arrays. PMID- 11421721 TI - Tricyclohexylphosphine derivatives of bis(2,6-difluorophenoxide)cadmium: a solution and solid-state NMR study. PMID- 11421723 TI - Synthesis and single-crystal characterization of the square planar complex [Ni[(SPPh(2)(2)N-S,S']*2THF. PMID- 11421722 TI - Structurally homologous beta- and meso-amidinium porphyrins. PMID- 11421724 TI - Bis(tetrahydrofuran)(2,7,12,17-tetrapropylpor- phycenato)iron(III) perchlorate: a pure intermediate spin complex. PMID- 11421725 TI - A new dehydrogeranylgeraniol antioxidant from Saururus cernuus that inhibits intracellular reactive oxygen species (ROS)-catalyzed oxidation within HL-60 cells. AB - A new triene, 12,13-dehydrogeranylgeraniol (1), was isolated from the aquatic plant Saururus cernuus and its structure determined spectroscopically. Compound 1 inhibits PMA-induced peroxide-catalyzed oxidation of 2',7' dichlorodihydrofluorescein dye (DCFH) by reactive oxygen species (ROS) within human promyelocyctic HL-60 cells. PMID- 11421726 TI - New bromoterpenes from the red alga Laurencia luzonensis. AB - Extraction of a sample of Laurencia luzonensis collected off the coast of Kudaka Island, Okinawa, yielded the known sesquiterpenes palisol (1), palisadin B (2), palisadin A (3), pacifigorgiol (4), and aplysistatin (5), together with five new bromosesquiterpenes, isopalisol (6), luzonensol (7), luzonensol acetate (8), luzonensin (9), and (3Z,6E)-1-bromo-2-hydroxy-3,7,11-trimethyldodeca-3,6,10 triene (10). In addition, a new bromoditerpene of unusual structure, 3 bromobarekoxide (11), possessing a seven-membered ring fused to trans-decalin, was isolated. PMID- 11421727 TI - New prenylated bi- and tricyclic phloroglucinol derivatives from Hypericum papuanum. AB - Five new prenylated tricyclic and three new bicyclic acylphloroglucinol derivatives have been isolated by bioactivity-guided fractionation of the petroleum ether extract of the dried aerial parts of Hypericum papuanum. The tricyclic compounds (1--5) were named papuaforins A--E. The bicyclic compounds were isolated together with their corresponding tautomers and were named hyperguinones A and B (6/6a,7/7a) and hyperpapuanone (8/8a), respectively. Their structures were elucidated on the basis of extensive 1D and 2D NMR experiments, as well as mass spectrometry. Furthermore, the cytotoxicity toward KB nasopharyngeal carcinoma cells and the antibacterial activity of the isolated compounds were determined. PMID- 11421728 TI - Chemical constituents from Drypetes littoralis. AB - Chemical investigation of Drypetes littoralis yielded three new tricyclic diterpenes, drypetenones A, B, and C (1--3), and one new xanthone (4). Spectral analyses and chemical correlations established the structures as 10S-12-hydroxy 11-methoxy-13-methylpodocarpa-1,5,8,11,13-pentaene-3,7-dione, (1), 10S-12-hydroxy 11-methoxy-13-methylpodocarpa-5,8,11,13-tetraene-3,7-dione (2), 10S-12-hydroxy 6,11-dimethoxy-13-methylpodocarpa-1,5,8,11,13-pentaene-3,7-dione (3), and 1 hydroxy-7-hydroxymethyl-6-methoxyxanthone (4). Complete (13)C NMR assignment of boehmenan D (5) is also made. PMID- 11421729 TI - Five new prenylated stilbenes from the root bark of Lonchocarpus chiricanus. AB - Besides the known compounds longistylines C (1), D (2), and 3,5-dimethoxystilbene (5), five new prenylated stilbenes, named chiricanines A--E (3, 4, 6--8), have been isolated from the root bark of Lonchocarpus chiricanus. Their structures were resolved on the basis of spectrometric methods including (1)H, (13)C, and 2D NMR experiments and mass spectrometry. Compound 3 was the only prenylated stilbene to demonstrate antifungal effects against Cladosporium cucumerinum. Four of the isolated compounds showed toxic properties against larvae of the yellow fever-transmitting mosquito Aedes aegypti. Compound 5 was found to be as potent as rotenone in larvicidal dilution tests. PMID- 11421730 TI - Somamides A and B, two new depsipeptide analogues of dolastatin 13 from a Fijian cyanobacterial assemblage of Lyngbya majuscula and Schizothrix species. AB - Somamides A (1) and B (2) were isolated from assemblages of the marine cyanobacteria Lyngbya majusculaand Schizothrix sp. from the Fijian Islands. These new depsipeptides are analogous in structure to the cyanobacterial metabolite symplostatin 2 (4) as well as dolastatin 13 (3), originally isolated from Dolabella auricularia, further demonstrating the cyanobacterial origin of the dolastatins. PMID- 11421731 TI - Trypanocidal withanolides and withanolide glycosides from Dunalia brachyacantha. AB - Two new withanolide glycosides, (20R,22R)-O-(3)-alpha-L-rhamnopyranosyl(1-->4) beta-D-glucopyranosyl-1 alpha,12 beta-diacetoxy-20-hydroxywitha-5,24-dienolide (3) and (20R,22R)-O-(3)-beta-D-xylopyranosyl(1-->3)-[beta-D-xylopyranosyl(1-->4)] beta-D-glucopyranosyl-1 alpha-acetoxy-12 beta,20-dihydroxywitha-5,24-dienolide (4), were isolated from the leaves and root of Dunalia brachyacantha. Their aglycones, (20R,22R)-1 alpha,12 beta-diacetoxy-3 beta,20-dihydroxywitha-5,24 dienolide (or 1 alpha,12 beta-diacetyldunawithagenine) and (20R,22R)-1 alpha acetoxy-3 beta,12 beta,20-trihydroxywitha-5,24-dienolide (or 1 alpha-acetyl-12 beta-hydroxydunawithagenine), are novel. The known 18-acetoxywithanolide D (1) and 18-acetoxy-5,6-deoxy-5-withenolide D (2) were also isolated from the leaves. These last two compounds were shown to be responsible for the trypanocidal, leishmanicidal, and bactericidal activities manifested by the crude ethanolic extract. The structures were deduced from spectroscopic data and on the basis of chemical evidence. PMID- 11421732 TI - Dendocarbins A--N, new drimane sesquiterpenes from the nudibranch Dendrodoris carbunculosa. AB - Fourteen new sesquiterpenes of the drimane series dendocarbins A--N (1--14) were obtained from ethanol extracts of the Japanese nudibranch Dendrodoris carbunculosa, together with two known compounds, isodrimeninol (15) and 11 epivaldiviolide (16). All structures were elucidated mainly from spectral data, and most of these sesquiterpenes were found to exhibit cytotoxicity. In addition, isodrimeninol (15), the major sesquiterpene of this animal, was found to have a sharp peppery taste. PMID- 11421733 TI - Two new cytotoxic and virucidal trisulfated triterpene glycosides from the Antarctic sea cucumber Staurocucumis liouvillei. AB - Two new trisulfated triterpene glycosides, liouvillosides A (1) and B (2), have been isolated from the Antarctic sea cucumber Staurocucumis liouvillei. Their structures have been elucidated by spectroscopic analysis (NMR and FABMS) and chemical transformations. Liouvillosides A (1) and B (2) are two new examples of a small number of trisulfated triterpene glycosides from sea cucumbers belonging to the family Cucumariidae. Both glycosides were found to be virucidal against herpes simplex virus type 1 (HSV-1) at concentrations below 10 microg/mL. PMID- 11421734 TI - Megastigmane glycosides and an acylated triterpenoid from Eriobotrya japonica. AB - Two new megastigmane glycosides, eriojaposides A (1) and B (2), and a new acylated triterpenoid (3) were isolated along with nine known compounds from a leaf extract of Eriobotrya japonica. The structures of 1--3 were characterized as (6R,9R)-3-oxo-alpha-ionyl-9-O-beta-xylopyranosyl-(1' '-->6')-beta glucopyranoside, (6R,9R)-3-oxo-alpha-ionyl-9-O-alpha-rhamnopyranosyl-(1' '-->6') beta-glucopyranoside, and 3 alpha-trans-feruloyloxy-2 alpha-hydroxyurs-12-en-28 oic acid, respectively, on the basis of spectral and chemical evidence. PMID- 11421735 TI - Gelliusterols A--D, new acetylenic sterols from a sponge, Gellius species. AB - New acetylenic sterols, gelliusterol A (1, 26,27-bisnorcholest-5-en-23-yn-3 beta,7 alpha-diol), its corresponding 7-ketone, gelliusterol B (2, 26,27 bisnorcholest-5-en-23-yn-3 beta-ol-7-one), and gelliusterols C (4, cholest-5-en 23-yn-3 beta,7-one) and D (5, cholest-5-en-23-yn-3 beta,25-diol-7-one), were isolated from an unidentified species of sponge, Gellius sp. The structures of the steroids were established from spectroscopic data. PMID- 11421736 TI - Cox-2 inhibitory effects of naturally occurring and modified fatty acids. AB - In the search for new cyclooxygenase-2 (COX-2) selective inhibitors, the inhibitory effects of naturally occurring fatty acids and some of their structural derivatives on COX-2-catalyzed prostaglandin biosynthesis were investigated. Among these fatty acids, linoleic acid (LA), alpha-linolenic acid (alpha-LNA), myristic acid, and palmitic acid were isolated from a CH(2)Cl(2) extract of the plant Plantago major by bioassay-guided fractionation. Inhibitory effects of other natural, structurally related fatty acids were also investigated: stearic acid, oleic acid, pentadecanoic acid, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). Further, the inhibitory effects of these compounds on COX-2- and COX-1-catalyzed prostaglandin biosynthesis was compared with the inhibition of some synthesized analogues of EPA and DHA with ether or thioether functions. The most potent COX-2-catalyzed prostaglandin biosynthesis inhibitor was all-(Z)-5-thia-8,11,14,17-eicosatetraenoic acid (2), followed by EPA, DHA, alpha-LNA, LA, (7E,11Z,14Z,17Z)-5-thiaeicosa-7,11,14,17-tetraenoic acid, all-(Z)-3-thia-6,9,12,15-octadecatetraenoic acid, and (5E,9Z,12Z,15Z,18Z)-3 oxaheneicosa-5,9,12,15,18-pentaenoic acid, with IC(50) values ranging from 3.9 to180 microM. The modified compound 2 and alpha-LNA were most selective toward COX-2, with COX-2/COX-1 ratios of 0.2 and 0.1, respectively. This study shows that several of the natural fatty acids as well as all of the semisynthetic thioether-containing fatty acids inhibited COX-2-catalyzed prostaglandin biosynthesis, where alpha-LNA and compound 2 showed selectivity toward COX-2. PMID- 11421737 TI - Triterpene saponins from Tupidanthus calyptratus. AB - Five new bisdesmosidic saponins (1--5) were isolated from the aerial parts of Tupidanthus calyptratus. Their structures were determined by (1)H--(1)H correlation spectroscopy (COSY, TOCSY, ROESY) and (1)H--(13)C correlation (HSQC, HMBC) NMR experiments, FABMS, and chemical data. PMID- 11421738 TI - Milolides, new briarane diterpenoids from the western Pacific octocoral Briareum stechei. AB - Eleven new briarane-type diterpene lactones, designated milolides (1--8, 10--12), together with four known diterpene lactones (9, 13--15), were isolated from the Micronesian octocoral Briareum stechei collected at Yap, Federated States of Micronesia. One aspect of the stereochemistry of one of the known compounds, solenolide C (9), is revised. Structures of the new compounds were determined from spectral data. PMID- 11421739 TI - Synthetic derivatives of abietic acid with radical scavenging activity. AB - In this work, studies on the arylation of anilines derived from dehydroabietic acid, the main component of disproportionated rosin, are presented. The redox properties of the new diarylamines were investigated by cyclic voltammetry, and their free radical scavenging activity was tested by reduction of the 2,2 diphenyl-1-picrylhydrazyl (DPPH) radical. Three of the diarylamines with lower oxidation potential proved to be as active as isopropyldiphenylamine (IPPD) and superior to tert-butylhydroxytoluene (BHT), both commercially available synthetic antioxidants. PMID- 11421740 TI - New triterpenoid saponins from the sponge Erylus nobilis. AB - Erylosides G--J (1--4), four new triterpenoid saponins, were isolated from the sponge Erylus nobilis collected from Jaeju Island, Korea. On the basis of the results of combined chemical and spectral analyses, the structures of the aglycones were determined to be lanostane-based, modified penasterols. The oligosaccharide portions were composed of one unit each of L-arabinose, D galactose, and 2-N-acetyl-D-glucosamine (1 and 3) or two units of L-arabinose and one unit of 2-N-acetyl-D-glucosamine (2 and 4). These compounds exhibited moderate cytotoxicty against a human leukemia cell line. PMID- 11421741 TI - Antimalarial compounds from Rhaphidophora decursiva. AB - Bioassay-directed fractionation led to the isolation of 14 compounds, six of which possess antimalarial activity, from the dried leaves and stems of Rhaphidophora decursiva. Polysyphorin (1) and rhaphidecurperoxin (6) showed strong activities against Plasmodium falciparum. Rhaphidecursinol A (2), rhaphidecursinol B (3), grandisin (4), and epigrandisin (5) were less active against the same organism. Among the isolates, rhaphidecursinol A (2) and rhaphidecursinol B (3) were determined to be new neolignans, and rhaphidecurperoxin (6) is a new benzoperoxide. Known compounds isolated include polysyphorin (1), grandisin (4), epigrandisin (5), (+)-medioresinol, (-) pinoresinol, (-)-syringaresinol, (+)-glaberide I, (+)-dehydrovomifoliol, (-) liliolide, (-)-hydroxydihydrobovolide, and N-butylbenzamide, of which compound 1 appears worthy of further evaluation as an antimalarial agent. Structure elucidation and identification were accomplished by spectroscopic means including 1D and 2D NMR analyses. PMID- 11421742 TI - Neoclerodane diterpenoids from Scutellaria caerulea. AB - Five new neoclerodane diterpenoids have been isolated from Scutellaria caerulea: (11S*)-6 alpha-acetoxy-7 beta,11-diisobutiryloxy-1 beta,8 beta-dihydroxy-4(18),13 neoclerodadien-15,16-olide (scuterulein A) (1); (13R*)-1 beta-6 alpha-7 beta triacetoxy-11 beta-benzoyloxy-8 beta,13-epoxy-4(18)-neocleroden-15,16-olide (scuterulein B) (2); (11S*)-1 beta,6 alpha,11-triacetoxy-7 beta-isobutiryloxy-8 beta-hydroxy-4(18),13-neoclerodadien-15,16-olide (scuterulein C) (3); (11S*)-6 alpha,11-diacetoxy-7 beta-isobutiryloxy-1 beta,8 beta-dihydroxy-4(18),13 neoclerodadien-15,16-olide (deacetyl scuterulein C) (4), and (11E)-6 alpha acetoxy-7 beta-isobutiryloxy-1 beta,8 beta-dihydroxy-4(18),11,13-neoclerodatrien 15,16-olide (scuterulein D) (5). Structures were established by spectroscopic and chemical methods. An X-ray analysis was carried out on scuterulein B (2). PMID- 11421743 TI - Withanolides from Salpichroa origanifolia. AB - Five new withanolides, 5 alpha,6 alpha:22,26:24,25-triepoxy-16 alpha,26-dihydroxy 18(13-->17)-abeo-ergosta-2,13-dien-1-one (salpichrolide N, 1), 5 alpha,6 alpha:22,26:24,25-triepoxi-15 alpha,26-dihydroxyergosta-2,16-dien-1-one (salpichrolide L, 2), 5 alpha,6 alpha:22,26-diepoxi-24,25,26-trihydroxy-17(13- >18)-abeo-ergosta-2,13,15,17-tetraen-1-one (salpichrolide M, 3a), 5 alpha,6 alpha:22,25:22,26-triepoxy-24-hydroxy-17(13-->18)-abeo-ergosta-2,13,15,17-tetraen 1-one (salpichrolide J, 4), and 5 alpha,6 alpha:22,26-diepoxy-22,24,25-trihydroxy 17(13-->18)-abeo-ergosta-2,13,15,17-tetraen-1-one (salpichrolide K, 5), were isolated from the leaves of Salpichroa origanifolia and characterized by a combination of spectroscopic (1D and 2D NMR, MS) and chemical methods. PMID- 11421744 TI - Norditerpenoid and diterpenoid alkaloids from Turkish Consolida orientalis. AB - From the aerial parts of Consolida orientalis collected in Turkey, a new diterpenoid alkaloid designated as consorientaline has been isolated along with the known norditerpenoid alkaloids delsoline, delcosine, gigactonine, and takaosamine. The structure of consorientaline (1) was established by spectroscopic studies and chemical correlation with dihydroajaconine (2). PMID- 11421745 TI - Ma'iliohydrin, a cytotoxic chamigrene dibromohydrin from a Philippine Laurencia species. AB - A new cytotoxic tribrominated chamigrene with a dibromohydrin functionality was isolated from an undetermined species of red alga, Laurencia sp. Its structure was determined by spectroscopic methods. PMID- 11421746 TI - Chemical constituents from Lippia sidoides and cytotoxic activity. AB - Eleven known compounds and a new prenylated naphthoquinone, lippsidoquinone (13), were isolated from ethanol extracts of Lippia sidoides. Their structures were established by a combination of 1D and 2D NMR, IR, and EIMS spectral data analysis. The cytotoxic properties of compounds 3--13 were evaluated against HL60, SW1573, and CEM cell lines. Only tectol (6) and lippsidoquinone (13) exhibited significant activity against human leukemia cell lines HL60 and CEM. PMID- 11421747 TI - Iridoids from Rothmannia macrophylla. AB - A new iridoid glucoside with an ether linkage between C-3 and C-10 and a novel nonglycosidic iridoid with an ether linkage between C-3 and C-6 and a lactonic linkage at C-1, named macrophylloside (1) and macrophyllide (2), respectively, were isolated from the leaves of Rothmannia macrophylla, along with six known iridoids. Their structures were established by NMR and MS spectroscopies. PMID- 11421748 TI - Iridoid glycosides from the leaves of Morinda citrifolia. AB - A new iridoid glucoside (1), named citrifolinoside A, was isolated from the leaves of Morinda citrifolia along with the known iridoids asperuloside and asperulosidic acid. The structure of 1 was established by interpretation and full assignments of NMR spectroscopic data. PMID- 11421749 TI - New sesquiterpene hydroquinones from a Taiwanese marine sponge, Hippospongia metachromia. AB - In addition to metachromins A (1), B (2), and E (9), two new sesquiterpene hydroquinones, hippochromins A and B (3, 4), were isolated from the Taiwanese marine sponge Hippospongia metachromia. The structures of 4--6 were established by extensive 2D NMR analysis. Metachromins A (1) and B (2), hippochromin A diacetate (5), and metachromin B monoacetate (8) exhibited potent cytotoxicities against human colon (COLO-205) and nasopharyngeal (KB) tumor cells, while compounds 6, 7, and 9 were inactive. PMID- 11421750 TI - Porwenins A and B, new clerodane diterpenoids from Portulaca okinawensis. AB - Two new clerodane-type diterpenes, porwenins A (1) and B (2), were isolated from Portulaca okinawensis, and the structures were elucidated by spectroscopic data. PMID- 11421751 TI - Verbenachalcone, a novel dimeric dihydrochalcone with potentiating activity on nerve growth factor-action from Verbena littoralis. AB - A novel dimeric dihydrochalcone, verbenachalcone (1), was isolated from the aerial parts of Verbena littoralis. Its structure was elucidated, on the basis of spectral data interpretation, as 4,2',4',2' ",4' "-pentahydroxy-3' '-methoxy-3-O 4' '-tetrahydrobichalcone. This compound caused a significant enhancement of nerve growth factor-mediated neurite outgrowth from PC12D cells. PMID- 11421752 TI - Bombardolides: new antifungal and antibacterial gamma-lactones from the coprophilous fungus Bombardioidea anartia. AB - Chemical studies of organic extracts from cultures of the coprophilous fungus Bombardioidea anartia have led to the discovery of bombardolides A--D (1--4), a series of new antifungal and antibacterial metabolites. Three of these metabolites (1--3) were obtained as inseparable pairs of geometric isomers. A new 3-substituted phenol (5) and the known compound asterriquinone B4 were also encountered. The structures of compounds 1--5 were determined by analysis of NMR and MS data. PMID- 11421753 TI - Stelliferin riboside, a triterpene monosaccharide isolated from the Fijian sponge Geodia globostellifera. AB - A triterpene monosaccharide, stelliferin riboside (1), was isolated from the Fijian marine sponge Geodia globostellifera. In addition, stellettins A and B (2, 3) were also isolated. This is the first report of a stelliferin from this species and the first example of a saccharide derivative of a stelliferin. PMID- 11421754 TI - Two new antimicrobial lysoplasmanylinositols from the marine sponge Theonella swinhoei. AB - Two new antimicrobial lysoplasmanylinositols have been isolated from the marine sponge Theonella swinhoei. Their structures were elucidated on the basis of spectroscopic and chemical analysis. PMID- 11421755 TI - Thalibealine, a novel tetrahydroprotoberberine--aporphine dimeric alkaloid from Thalictrum wangii. AB - A novel tetrahydroprotoberberine-aporphine dimeric alkaloid, (-)-thalibealine (1), was isolated from the roots of Thalictrum wangiii, and its structure established via spectroscopic analysis. Three other alkaloids were isolated, including the benzyltetrahydroisoquinoline-aporphine dimer (+)-thalmelatidine, the aporphine (+)-magnoflorine, and the protoberberine berberine. This is the first reported isolation of a tetrahydroprotoberberine-aporphine dimer from nature, as well as the first reported isolation of constituents from Thalictrum wangii. PMID- 11421756 TI - Synthesis of thalprzewalskiinone, a revision of structure. AB - A direct comparison of the spectral data for synthetic 2-methyl-6,7-dimethoxy-3' methoxy-4'-hydroxyoxobenzylisoquinoline iodide (1) and its positional isomer 2 methyl-6,7-dimethoxy-3'-hydroxy-4'-methoxyoxobenzylisoquinoline iodide (2) with the data obtained for the oxobenzylisoquinoline alkaloid thalprzewalskiinone revealed that the original structural assignment of the alkaloid as 1 was in error. These results mandate the revision of structure of thalprzewalskiinone to 2-methyl-6,7-dimethoxy-3'-hydroxy-4'-methoxyoxobenzylisoquinoline iodide (2). PMID- 11421757 TI - Allelopathic prenylflavanones from the fallen leaves of Macaranga tanarius. AB - Two new prenylflavanones, tanariflavanones A (1), and B (2), and one known compound, (-)-nymphaeol-C (3), were isolated from the fallen leaves of Macaranga tanarius. Compounds 1 and 2 showed inhibition of radical growth of lettuce seedlings at 200 ppm. Their structures were elucidated primarily by NMR, circular dichroism, and mass spectroscopic methods. PMID- 11421758 TI - New diterpenes from Jatropha divaricata. AB - Extracts of the stems of Jatropha divaricata have yielded the two new diterpenes ent-3 beta,14 alpha-hydroxypimara-7,9(11),15-triene-12-one (3) and the rearranged pimarane ent-15(13-->8)abeo-8 beta(ethyl)pimarane (4), which appears to be a new skeletal type. The rare cleistanthane diterpenes spruceanol (1) and cleistanthol (2) were also obtained. PMID- 11421759 TI - New dihydrophenanthrene and phenyldihydroisocoumarin constituents of Trema orientalis. AB - In the course of the chemical investigation of extracts of the trunk bark and root bark of Trema orientalis, three new compounds were isolated, namely, (9S*,10S*)-3-[7-(3,10-dihydroxy-9-hydroxymethyl-2,5-dimethoxy)-9,10 dihydrophenanthrenyl]propenal (1), (9S*,10S*)-3-[7-(5-O-beta-glucopyranosyl-10 hydroxy-9-hydroxymethyl-2,6-dimethoxy)-9,10-dihydrophenanthrenyl]propenal (2), and (3R*,3aR*,4R*,5S*)-6-O-alpha-arabinopyranosyl-8-hydroxy-3-(4-hydroxyphenyl)-4 (4-hydroxyphenyl)-5-(3,5-dihydroxyphenyl)-3,3a-dihydrocyclopenta[1,2,3 de]isobenzopyran-1-one (3, orientoside A). The structures of 1-3 were determined by spectral methods. PMID- 11421760 TI - Dihydrocarbazole alkaloids from Aspergillus tubingensis. AB - Investigation of the fungus Aspergillus tubingensis has led to the isolation and identification of two dihydrocarbazole-containing compounds (1 and 2). Details of the purification and structure elucidation of 1 and 2 are described. This is the first known report of dihydrocarbazole-containing compounds to be isolated from a living system. PMID- 11421761 TI - Alkaloids of the flowers of Hippeastrum vittatum. AB - Investigation of an ethanolic extract of the fresh flowers of Hippeastrum vittatum yielded the new alkaloids vittacarboline (1) and O-methylismine (3), together with the known compound ismine (2). The structures of 1--3 were established by spectroscopic methods including one- and two-dimensional NMR and mass spectrometry. PMID- 11421762 TI - Steroidal alkaloids from the leaves of Sarcococca coriacea of Nepalese origin. AB - Two new steroidal alkaloids, (-)-vaganine D (1) [(20S)-20-(N,N-dimethylamino)-3 beta-(senecioylamino)-5 alpha-pregn-16-en-4 beta-yl acetate], and (+) nepapakistamine A (2) [(20S)-20-(N-methylamino)-3 beta-(tigloylamino)-5 alpha pregn-16-en-2 beta,4 beta-diacetate], were isolated from the leaves of Sarcococca coriacea. Their structures were elucidated on the basis of their spectral properties. Compounds 1 and 2 were found to be cholinesterase inhibitors. PMID- 11421763 TI - Bromolysis and iodolysis of alpha,beta-epoxycarboxylic acids in water catalyzed by indium halides. AB - The ring opening of alpha,beta-epoxycarboxylic acids by bromide and iodide ions has been efficiently carried out in water in high regio- and stereoselective fashion. The iodolysis of trans-beta-monoalkylated epoxycarboxylic acids at pH 4.0 was completely alpha-regioselective and anti diastereoselective. The InCl(3) catalyzed iodolysis of a variety of alpha,beta-epoxycarboxylic acids at pH 1.5 gave the corresponding anti beta-iodohydrins in 88-95% yields. The one-pot synthesis of the alpha- and beta-hydroxyhexanoic acids, starting from the corresponding alpha,beta-epoxycarboxylic acid 1a by iodolysis followed by reduction of the resulting iodohydrins 4a and 4b by NaBH(4)-InCl(3) in water, has been performed. PMID- 11421764 TI - Engineering reactions in crystalline solids: photochemical generation of secondary and tertiary enol radical pairs from crystalline ketodiesters. AB - The photochemical decarbonylation of several crystalline 1,3 acetonedicarboxylates has been analyzed in solution and in the solid state. It is shown that the efficiency of the solid-state reaction depends on the stability of the intermediate acyl-alkyl and alkyl-alkyl radical pairs. Reactions proceeding through tertiary enol radicals are more efficient than reactions proceeding through secondary enol radical centers. Solid-state reactions that require the intermediacy of primary enol radicals do not occur. It is also shown that the selectivity of product formation in crystals depends on the structure of the reactant solid phase. PMID- 11421765 TI - A strategy for the assembly of multiple porphyrin arrays based on the coordination chemistry of Ru-centered porphyrin pentamers. AB - An approach which employs pentameric porphyrin arrays as building blocks toward larger porphyrin arrays is described. Two flexible, and one relatively rigid, Ru centered porphyrin pentamers (1-3) were synthesized and fully characterized. Their potential as building blocks toward larger porphyrin arrays has been studied via their coordination chemistry using bidentate and tetradentate ligands. DABCO (diazabicyclo[2.2.2]octane) can bind two monomeric porphyrins but was found to be too small to allow the complete formation of a 10-porphyrin array. On the other hand, titration of a larger bridging dipyridyl porphyrin ligand 17 (0.5 equiv) with 1 or 2 and tetrapyridyl ligand 18 (0.25 equiv) with 3 results in the formation of the 11-porphyrin and 21-porphyrin arrays, respectively, with the 21-porphyrin array containing porphyrins in three different metalation states. Changes in the chemical shift of the inner NH protons as well as the ortho- and meso-protons of the pyridyl groups of the porphyrin ligand clearly indicate the formation of large multiple porphyrin complexes. These studies demonstrate that by use of carefully designed building blocks and suitable bridging ligands, porphyrin arrays can be constructed with a dramatic increase in size in relatively few steps. Exploiting the fact that the strength of binding of pyridyl ligands is Ru > Zn > Ni, intra- vs intermolecular competition has been used to investigate aspects of the folding of the array. The photophysical properties of 3 are also described. PMID- 11421766 TI - Mechanism of electroreduction of allyl alcohol at platinized platinum electrode in acidic aqueous solution. AB - The electroreduction of allyl alcohol to form propene at the platinized platinum electrode in acidic aqueous solution has been studied using CV plots, IR, ESR, and MS spectra, and a semiempricial MO method (MOPAC7/AM1, PM3). From the determinations of charge-transfer coefficients, reaction orders and apparent activation energy for the given reaction, the detection of the intermediates such as C(3)H(5)(+), C(3)H(5)(*), and C(3)H(5)(-) species, and PM3 calculations of charge distribution and frontier orbital energies of the reaction species C(3)H(5)OH and C(3)H(5)(+), the authors suggest that in acidic aqueous solution the production of propene via reductive splitting of the C-OH bond situated in the allyl position of allyl alcohol obeys a carbonium ion-carbanion mechanism. PMID- 11421767 TI - Synthesis of a series of oligo(ethylene glycol)-terminated alkanethiol amides designed to address structure and stability of biosensing interfaces. AB - A strategy for the synthesis of a series of closely related oligo(ethylene glycol)-terminated alkanethiol amides (principally HS(CH(2))(m)CONH(CH(2)CH(2)O)(n)H; m = 2, 5, 11, 15, n = 1, 2, 4, 6, 8, 10, 12) and analogous esters has been developed. These compounds were made to study the structure and stability of self-assembled monolayers (SAMs) on gold in the prospect of designing new biosensing interfaces. For this purpose, monodisperse heterofunctional oligo(ethylene glycols) with up to 12 units were prepared. Selective monoacylation of the symmetrical tetra- and hexa(ethylene glycol) diols as their mesylates with the use of silver(I) oxide was performed. The synthetic approach was based on carbodiimide couplings of various oligo(ethylene glycol) derivatives to omega-(acetylthio) carboxylic acids via a terminal amino or hydroxyl function. SAM structures on gold were studied with respect to thickness, wettability (water contact angles approximately 30 degrees ), and conformation. A good fit was obtained for the relation between monolayer thickness (d) and the number of units in the oligo(ethylene glycol) chain (n): d = 2.8n + 21.8 (A). Interestingly, the corresponding infrared spectroscopy analysis showed a dramatic change in conformation of the oligomeric chains from all-trans (n = 4) to helical (n > or = 6) conformation. A crystalline helical structure was observed in the SAMs for n > 6. PMID- 11421768 TI - Unexpected catalyzed C=C bond cleavage by molecular oxygen promoted by a thiyl radical. AB - Olefin oxidation with molecular oxygen, promoted by a transition metal catalyst and a thiophenol, involved C=C bond cleavage into the corresponding carbonyl derivatives. This new reaction proceeds under one atmosphere of oxygen, at room temperature, in the presence of an excess of thiophenol and a catalyst such as MnL(2) 3a or VClL(2) 3c. It was applied to aromatic and aliphatic olefins, as well as to functionalized or unfunctionalized acyclic compounds, providing the corresponding ketones and aldehydes in up to 98% yield. The synthetic interest of this catalytic oxidation was illustrated by a one-step preparation of the fragrance (-)-4-acetyl-1-methylcyclohexene 7e in 73% isolated yield. The C=C bond cleavage probably results from a catalyzed decomposition of the beta hydroperoxysulfide intermediate 12 that is formed by the radical addition of thiophenol to the olefin in the presence of oxygen. Although an excess of the thiophenol was used, it was transformed into the disulfide which could then be reduced without purification in 83% overall yield, thereby allowing for recycling. In addition, the C=C bond cleavage under oxygen could be promoted by catalytic quantities of the thiyl radical, generated by photolysis of the disulfide; thus, in the presence of 0.1 equiv of bis(4-chlorophenyl) disulfide 4b and 5% of the manganese complex 3a, trans-methylstilbene 1b gave, under radiation, benzaldehyde 6a and acetophenone 7a in up to 95% yield. This new reaction offers an alternative to the classical C=C bond cleavage procedures, and further developments in the fields of bioinorganic and environmental chemistry are likely. PMID- 11421769 TI - Sequenced reactions with samarium(II) iodide. Sequential intramolecular Barbier cyclization/Grob fragmentation for the synthesis of medium-sized carbocycles. AB - Samarium(II) iodide was used to access eight-, nine-, and ten-membered carbocycles via a domino reaction composed of a cyclization/fragmentation process. 2-(Iodoalkyl)-, 2-(iodomethyl)allyl-, and 2-(2-iodomethyl)benzyl-2 methyl-3-(methanesulfonyloxy)cycloalkanones were subjected to Barbier-type reductive coupling conditions. Intermediate cycloalkanedione derivatives were also treated under similar conditions, providing bicyclic hydroxy ketones with complete diastereoselectivity and high yields. This method represents a general and efficient approach to a variety of highly functionalized, stereodefined carbocycles. PMID- 11421770 TI - Trialkyltetrathiafulvalene-sigma-tetracyanoanthraquinodimethane [R(3)TTF-sigma TCNAQ] diads: synthesis, intramolecular charge-transfer properties, and X-ray crystal structure. AB - We report the use of the electron-donating 4,5-dipentyl-4'-methyl-TTF (TTF = tetrathiafulvalene) moiety in combination with the electron acceptor 11,11,12,12 tetracyanoanthraquinodimethane (TCNAQ) unit in the novel D-sigma-A diad molecules 11, 17, and 18. These compounds display a weak, broad, low-energy intramolecular charge-transfer (ICT) band in the UV-vis spectra (lambda(max) 430-450 nm). Cyclic voltammetric studies show two reversible one-electron oxidation processes for the R(3)TTF moiety, and a reversible two-electron reduction process for the TCNAQ moiety. The electron affinity of TCNAQ is significantly enhanced by the electron withdrawing sulfonamide and sulfonic ester groups (compounds 17 and 18, respectively). Simultaneous electrochemistry and EPR (SEEPR) experiments show no significant intramolecular interaction between the R(3)TTF and TCNAQ moieties in compounds 11 and 18. X-ray crystallographic data are presented for 5, 11, and 20. The structure of 5 reveals hydrogen-bonded dimers. In molecule 11 the bond lengths and conformations of both donor and acceptor moieties are typical for neutral species. Compound 20 is an unusual calcium complex of TCNAQ derivative obtained by dicyanomethylation of anthraquinone-2-carboxylic acid. PMID- 11421771 TI - Efficient asymmetric synthesis of biologically important tryptophan analogues via a palladium-mediated heteroannulation reaction. AB - A novel and concise synthesis of optically active tryptophan derivatives was developed via a palladium-catalyzed heteroannulation reaction of substituted o iodoanilines with an internal alkyne. The required internal alkyne 14a or 25 was prepared in greater than 96% de via alkylation of the Schollkopf chiral auxiliary 19 employing diphenyl phosphate as the leaving group. The Schollkopf chiral auxiliary was chosen here for the preparation of L-tryptophans would be available from D-valine while the D-isomers required for natural product total synthesis would originate from the inexpensive L-valine (300-g scale). Applications of the palladium-catalyzed heteroannulation reaction were extended to the first asymmetric synthesis of L-isotryptophan 38 and L-benz[f]tryptophan 39. More importantly, the optically pure 6-methoxy-D-tryptophan 62 was prepared by this protocol on a large scale (>300 g). This should permit entry into many ring-A oxygenated indole alkaloids when coupled with the asymmetric Pictet-Spengler reaction. In addition, an improved total synthesis of tryprostatin A (9a) was accomplished in 43% overall yield employing this palladium-mediated process. PMID- 11421772 TI - Bis(acylsilanes) and trifluoromethyltrimethylsilane: a useful system for the synthesis of cyclic 2,2-difluoro-3-trialkylsilyl-1,3-ketols and cyclic 2-fluoro 1,3-diketones(1). AB - We report a one-pot reaction of bis(acylsilanes) with trifluoromethyltrimethylsilane (TFMTMS) leading to a new family of 2,2-difluoro-3 trialkylsilylketols 4. These compounds were submitted to a facile and effective defluorosilylation. The overall process constitutes a new synthesis of cyclic six and seven-membered 2-fluoro-1,3-diketones 8, with regiospecific introduction of fluorine. The keto-enol equilibrium of cyclic 1,3-diketones and the mechanism of the defluorosilylation reaction were also studied. PMID- 11421773 TI - Sensitivity of (1)J[C(1)-H(1)] magnitudes to anomeric stereochemistry in 2,3 anhydro-O-furanosides. AB - The magnitude of the one-bond coupling constant between C(1) and H(1) in 2,3 anhydro-O-furanosides has been shown to be sensitive to the stereochemistry at the anomeric center. A panel of 24 compounds was studied and in cases where the anomeric hydrogen is trans to the epoxide moiety, (1)J[C(1)-H(1)] = 163-168 Hz; and when this hydrogen is cis to the oxirane ring, ((1)J[C(1)-H(1)] = 171-174 Hz. In contrast, for 2,3-anhydro-S-glycosides, the size of the (1)J[C(1)-H(1)] is not sensitive to C(1) stereochemistry. Computational studies on all four methyl 2,3 anhydro-O-furanosides (5-8) demonstrated that (1)J[C(1)-H(1)] was inversely proportional to the length of the C(1)-H(1) bond. A previously reported equation, which relates C(1)-H(1) bond distance and atomic charges to (1)J[C(1)-H(1)] magnitudes, could be used to accurately predict the J values in the alpha-lyxo (5) and beta-ribo (8) isomers. In contrast, with the beta-lyxo (6) and alpha-ribo isomers (7), this equation underestimated the size of these coupling constants by 10-20 Hz. PMID- 11421774 TI - Facile enzymatic aldol reactions with dihydroxyacetone in the presence of arsenate. AB - Aldol reactions of in situ formed dihydroxyacetone arsenate with different aldehydes were catalyzed by bacterial D-fructose-1,6-bisphosphate aldolase (FruA). Aldolases from bacteria were found to be much more stable and active than FruA from rabbit muscle. Arsenate acts as a phosphate mimic and can, in principle, be used in catalytic amounts. The use of inorganic arsenate and dihydroxyacetone afforded high yields with hydrophobic aldehydes. Cosolvents increased the solubility of hydrophobic aldehydes and afforded higher reaction rates and enzyme stability. Insight is given, for the first time, in the influence of arsenate on the stereoselectivity of the aldol reaction. PMID- 11421775 TI - Palladium-copper catalyzed synthesis of benzofused heterocycles with two heteroatoms: novel and highly regio- and stereoselective syntheses of (E)-2-(2 arylvinyl)-3-tosyl-2,3-dihydro-1,3-benzothiazoles and (E)-2-alkyl(aryl)idene-3,4 dihydro-2H-1,4-benzothiazines. AB - A highly novel, general, and convenient palladium and copper-catalyzed procedure has been developed for the synthesis of (E)-2-(2-arylvinyl)-3-tosyl-2,3-dihydro 1,3-benzothiazoles 28-40. 3-(2-Aminophenylthio)prop-1-yne 1 reacts with aryl iodides 2-14 under palladium-copper catalysis to yield the disubstituted alkynes 15-27 which after tosylation undergo a novel cyclization with CuI in the presence of triethylamine in THF to (E)-2-(2-arylvinyl)-3-tosyl-2,3-dihydro-1,3 benzothiazoles 28-40 rather than to the expected 3-alkylidene-4-tosyl-3,4-dihydro 2H-1,4-benzothiazines 41. The reaction is highly regio- and stereoselective. The synthesis of 2-(2-arylethyl)-3-tosylbenzothiazolines 42-47, 2-(2 arylvinyl)benzothiazoles 48-54, and a novel 5-substituted uracil derivative 55 of potential biological importance is also being reported. Similarly, the palladium copper-catalyzed arylation of S-[2-(N-prop-2'-ynyl)aminophenyl]-N,N dimethylthiocarbamate 58 with aryl iodides yields the disubstituted alkynes 59 which on cyclization with KOH in methanol leads to (E)-2-(2-aryl)methylidene-3,4 dihydro-2H-1,4-benzothiazines 61. The reaction of the diiodo compounds 12-14a, however, with 58 under palladium-copper-catalyzed reactions involves the participation of only one of the iodo groups in the heteroannulation process giving compounds 61i and 61j. These are amenable to further palladium-catalyzed reactions and afford polyunsaturated heteroaromatic compounds 62 and 63. PMID- 11421776 TI - Electrophilic oxidant produced in the photodeoxygenation of 1,2-benzodiphenylene sulfoxide. AB - We report that the photodeoxygenation of 1,2-benzodiphenylene sulfoxide, 1, generates an intermediate capable of oxidizing the solvent benzene to phenol. The reactivity of the intermediate was probed with various substrates (2 methylbutane, chloride ion, and para-substituted aryl sulfides). The intermediate produced in the sulfoxide photodeoxygenation displays an electrophilic oxidation chemistry. Our data on 1 contrast with the behavior of hydroxyl radical but resemble the chemistry observed for gas-phase atomic oxygen [O((3)P)] and for solution-phase photodeoxygenations of dibenzothiophene sulfoxide, 3, and pyridine N-oxide, 5. Correlations are made between the ionization potential of the acceptor molecules and the logarithm of the relative rate constants in order to advance the idea that the oxidizing agent of the title reaction may be solution phase O((3)P). PMID- 11421777 TI - Conformational analysis of dehydrodidemnin B (aplidine) by NMR spectroscopy and molecular mechanics/dynamics calculations. AB - Dehydrodidemnin B (DDB or aplidine), a potent antitumoral natural product currently in phase II clinical trials, exists as an approximately 1:1 mixture of two slowly interconverting conformations. These are sufficiently long-lived so as to allow their resolution by HPLC. NMR spectroscopy shows that this phenomenon is a consequence of restricted rotation about the Pyr-Pro(8) terminal amide bond of the molecule's side chain. The same technique also indicates that the overall three-dimensional structures of both the cis and trans isomers of DDB are similar despite the conformational change. Molecular dynamics simulations with different implicit and explicit solvent models show that the ensembles of three-dimensional structures produced are indeed similar for both the cis and trans isomers. These studies also show that hydrogen bonding patterns in both isomers are alike and that each one is stabilized by a hydrogen bond between the pyruvyl unit at the terminus of the molecule's side chain and the Thr(6) residue situated at the junction betwen the macrocycle and the molecule's side chain. Nevertheless, each conformational isomer forms this hydrogen bond using a different pyruvyl carbonyl group: CO(2) in the case of the cis isomer and CO(1) in the case of the trans isomer. PMID- 11421778 TI - Peptides constrained by an aliphatic linkage between two C(alpha) sites: design, synthesis, and unexpected conformational properties of an i,(i + 4)-linked peptide. AB - A novel route for the synthesis of cyclic peptides constrained by an aliphatic bridge between two C(alpha)sites, using a triply orthogonal protecting group strategy, is described. The synthesis of the orthogonally protected bis-amino acid 1, via an enantioselective route utilizing the Schollkopf and Evans methodologies, is first described. This is then incorporated into a short, alanine-rich peptide 13, using a novel triply orthogonal protecting group strategy to couple first one, then the other, amino acid moiety in such a way that an aliphatic bridge is formed between the i and i + 4 positions. Unexpectedly, the resulting constrained peptide does not adopt a helical conformation: instead, it is shown by CD at low temperature to adopt a left handed type II beta-turn conformation in aqueous media and a right-handed type I beta-turn conformation in TFE. PMID- 11421779 TI - General approach to the synthesis of persubstituted hydrophilic and amphiphilic beta-cyclodextrin derivatives. AB - Heptakis(2,3,6-tri-O-allyl)-beta-cyclodextrin 2 was converted to heptakis[2,3,6 tri-O-(2,3-dihydroxypropyl)]-beta-cyclodextrin 3 by osmium tetroxide-catalyzed dihydroxylation. A diastereomeric mixture of 3 was treated with sodium periodate followed by sodium borohydride to give heptakis[2,3,6-tri-O-(2-hydroxyethyl)] beta-cyclodextrin 5 in 86% yield. Compound 5 could be quantitatively transformed into heptakis(2,3,6-tri-O-carboxymethyl)-beta-cyclodextrin 6 by TEMPO-mediated oxidation. The same reaction sequence was also applied to heptakis(2,6-di-O-allyl 3-O-methyl)-beta-cyclodextrin 8, heptakis(2,3-di-O-allyl-6-O-methyl)-beta cyclodextrin 12, and heptakis(2,3-di-O-allyl-6-O-butyl)-beta-cyclodextrin 16; the analogous corresponding hydroxyethyl and carboxymethyl derivatives were isolated in high yields. All products were proved to be chemically uniform. PMID- 11421780 TI - Studies on porphyrin-quinhydrone complexes: molecular recognition of quinone and hydroquinone in solution. AB - Free-base and zinc(II) porphyrins bearing either one, two, or four hydroquinone entities at the meso positions are shown to bind quinones in solutions via a quinhydrone pairing mechanism. Electrochemical studies reveal that the quinhydrone complexes are stabilized by charge-transfer interactions between the donor (hydroquinone) and the acceptor (quinone). The redox potentials of the quinhydrone complexes are governed by the potentials of the quinones utilized to form quinhydrone. The (1)H NMR studies reveal that the quinhydrone complexes are stabilized by H-bonding in addition to the charge-transfer interactions. Singlet emission studies have shown that the fluorescence quenching of the porphyrin increases with an increase in the number of receptors, i.e., hydroquinone entities on the porphyrin macrocycle. Control experiments performed by using zinc porphyrin bearing a dimethoxyphenyl group, i.e., a receptor entity with no H bonding ability, indicate that the H-bonding plays an important role in quinhydrone formation. Porphyrin-quinhydrone formed by using covalently linked porphyrin-quinone and hydroquinone present in solution shows fluorescence enhancement. The measured fluorescence quantum yields, phi(f), are found to depend on the metal ion in the porphyrin cavity and the oxidation potential of the employed hydroquinones. The present studies also reveal that the measured phi(f) values depend on how the quinhydrone is linked to the porphyrin macrocycle, i.e., either through quinone or hydroquinone. Generally, porphyrin quinhydrone formed by hydroquinone-appended porphyrins shows decreased phi(f) values as compared to porphyrin-quinhydrone formed by quinone-appended porphyrins. PMID- 11421781 TI - Synthesis of novel acetal thia-cage compounds. AB - The synthesis of novel acetal thia-cage compounds has been accomplished by the direct substitution for the oxygen atom by the sulfur atom in the reaction of the acetal groups of oxa-cages with Lawesson reagent (LR). Reaction of the tetraoxa cage compound 2 with LR in dichloromethane at 25 degrees C sequentially gave the monothia-, dithia-, trithia-, and tetrathia-cage compounds 3, 6, 7, and 9. The reaction mechanism for the conversion from oxa-cages into thia-cages was proposed. The diacetal trioxa-cages 18-20 and 24-26 were also transformed into the thia-cages 21-23 and 27-29, respectively. Reaction of the trioxa-cages 34 and 35 with LR under the same reaction conditions gave the thia-cages 36 and 37 with the carbonyl group intact. Treatment of the pentaoxa[5]peristylane 40 with LR in chloroform under supersonic shaking at refluxing temperature gave the monothia[5]peristylane 41 and the dithia[5]peristylane 42. Attempts to synthesize the thia[5]peristylanes from the tetraoxa-cage 51 and the transformation from the parent (unsubstituted) pentaoxa[5]peristylane 46 to the thia-cages have been made. Reaction of the pentaoxa[5]-peristylane 40 with P(2)S(5) in refluxing toluene gave 41, 42, and a rearrangement product 47. The synthesis of new heterocyclic cage compounds 59 and 60, which contain oxygen, nitrogen, and sulfur atoms in the same molecule, was also accomplished. PMID- 11421782 TI - Kinetic resolution of acyclic secondary allylic silyl ethers catalyzed by chiral ketones. AB - Kinetic resolution of acyclic secondary allylic silyl ethers by chiral dioxiranes generated in situ from chiral ketones (R)-1 and (R)-2 and Oxone was investigated. An efficient and catalytic method has been developed for kinetic resolution of those substrates with a CCl(3), tert-butyl, or CF(3) group at the alpha-position. In particular, high selectivities (S up to 100) were observed for kinetic resolutions of racemic alpha-trichloromethyl allylic silyl ethers 7 and 9-15 catalyzed by ketones (R)-2. Both the recovered substrates and the resulting epoxides were obtained in high enantiomeric excess. On the basis of steric and electrostatic interactions between the chiral dioxiranes and the racemic substrates, a model was proposed to rationalize the enantioselectivities and diastereoselectivities in the chiral ketone-catalyzed kinetic resolution process. PMID- 11421783 TI - A convenient route to higher sugars by two-carbon chain elongation using Wittig/dihydroxylation reactions. AB - The combination of a Wittig olefination and a dihydroxylation reaction constitutes a facile synthetic protocol for the transformation of unprotected carbohydrates into higher sugars. The Wittig reaction is carried out with tert butyl or diphenylmethyl ester stabilized phosphoranes to give (E)-configured alpha,beta-unsaturated esters as the only products in most cases. These are dihydroxylated in a diastereoselective fashion using OsO(4)/NMO. The stereochemical outcome in the dihydroxylation follows Kishi's empirical rule and gives high diastereoselectivity (5:1-8:1) when starting from sugars with the 2,3 threo configuration. When starting from sugars with the 2,3-erythro configuration, the diastereoselectivity in the dihydroxylation is low (2:1 2.5:1). As a result, the Wittig/dihydroxylation protocol is most effective for producing higher sugars with the galacto configuration at the reducing end. The two steps can either be carried out individually or, more efficiently, as a one pot procedure. PMID- 11421784 TI - Efficient synthesis of enantiopure conduritols by ring-closing metathesis. AB - Two short synthetic approaches to enantiopure conduritols are described starting from the chiral pool. In both cases, the cyclohexene ring is assembled via ring closing olefin metathesis. The terminal diene precursers for the metathesis reaction are prepared either from octitols or from tartaric acids. The former route involves a new method for selective bromination of the primary positions in long-chain carbohydrate polyols. Subsequent reductive elimination with zinc then generates the diene. The latter route uses a highly diastereoselective addition of divinylzinc to tartaric dialdehydes for preparation of the dienes. PMID- 11421785 TI - Novel carbon-carbon bond-forming reactions using carbocations produced from substituted propargyl silyl ethers by the action of TMSOTf. AB - Highly useful carbon-carbon bond forming reactions using stable allenyl, propargyl, or allyl-propargyl hybrid cations have been developed. These carbocations could be generated from silyl 1-(pi-donor)-substituted propargyl ethers by the action of trimethylsilyl trifluoromethanesulfonate in dichloromethane at -78 degrees C to room temperature and could be attacked nucleophilically by electron rich arenes, allylsilanes, or enol silyl ethers, giving rise to allenes, alkynes, and their derivatives. A novel method for regio- and stereoselective synthesis of conjugated enynes utilizing allyl-propargyl hybrid cations has also been established. PMID- 11421786 TI - Convergent synthesis of noncovalent metallodendrimers containing hydrophobic dendrons at the periphery. AB - The noncovalent synthesis of "layer-block" metallodendrimers containing hydrophobic shells of covalent dendritic wedges at the periphery is described. Starting from first- and second-generation Frechet wedges having phosphines at their focal point, convergent dendritic growth yields third- and fourth generation metallodendrimers in which the coordination of nitriles, pyridines, and phosphines to SCS Pd(II) pincers is used as the assembly motif. In this convergent growth, the number of terminal hydrophobic phosphine wedges increases with generation. The solubility of the dendritic structures in apolar organic solvents such as chloroform and dichloromethane increases accordingly, in contrast to previously reported metallodendrimers. All dendritic structures were characterized by (1)H and (31)P NMR spectroscopy, elemental analysis, and MALDI TOF mass spectrometry. PMID- 11421787 TI - Nickel(0)-catalyzed fluoroalkylation of alkenes, alkynes, and aromatics with perfluoroalkyl chlorides. AB - Treatment of perfluoroalkyl chlorides (R(F)Cl) with alkenes, alkynes, or aromatics in the presence of 0.1 equiv of nickel dichloride, 1.5 equiv of zinc powder, and 0.4 equiv of triphenylphosphine in DMF at 95-100 degrees C for 6-8 h give the corresponding perfluoroalkylated products in good yields. A single electron-transfer mechanism is suggested. PMID- 11421788 TI - Synthesis of oligosaccharides of motifs D and E of arabinogalactan present in Mycobacterium tuberculosis. AB - Syntheses of the ethyl glycosides of 5-O-(beta-D-galactofuranosyl)-beta-D galactofuranose and 5-O-(alpha-D-arabinofuranosyl)-6-O-(beta-D-galactofuranosyl) beta-D-galactofuranose present in motifs D and E of Mycobacterium tuberculosis arabinogalactan, respectively, have been presented. The pentenyl-mediated O glycosylation reaction was utilized to obtain the disaccharide of motif D. The first coupling reaction to prepare the inner disaccharide portion of motif E was accomplished by trichloroacetamidate method while the installation of the terminal sugar by pentenyl glycosylation approach was successful. PMID- 11421789 TI - [4 + 2] cycloadditions of nitroalkenes in water. Highly asymmetric synthesis of functionalized nitronates. AB - The [4 + 2] cycloadditions of (E)-2-aryl-1-cyano-1-nitroalkenes 1 with achiral and enantiopure vinyl ethers 2 and 3 carried out in sole water are reported. These reactions occur in a heterogeneous phase under mild conditions and are fast and highly stereoselective. By using (-)-N,N-dicyclohexyl-(1S)-isoborneol-10 sulfonamide as a chiral auxiliary, the cycloadditions are totally asymmetric. The face selectivity is discussed in terms of the shape of the chiral auxiliary and the reactive conformation of vinyl ether. PMID- 11421790 TI - Facile and stereoselective access to nonracemic tricyclic cyclobutanes by asymmetric intramolecular Michael-aldol reaction: thermodynamic equilibrium and activation by iodonium ion. AB - Intramolecular Michael-aldol reactions of (-)-phenylmenthyl enoates tethered to cycloalkanone affords tricyclic cyclobutanes with high degrees of diastereochemical control. Kinetic and thermodynamic studies revealed that the Michael-aldol reaction is reversible under conditions in which trimethylsilyl iodide is used in the presence of hexamethyldisilazane at ambient temperature. Different levels of diastereoselectivity are observed when this cyclization process is carried out under kinetic vs thermodynamic conditions. Finally, an influence of added iodonium donors on the reaction rate has been noted. PMID- 11421791 TI - Biosynthesis of lipstatin. Incorporation of multiply deuterium-labeled (5Z,8Z) tetradeca-5,8-dienoic acid and octanoic acid. AB - Fermentation experiments with Streptomyces toxytricini were performed using (5Z,8Z)-[10,11,12,12-(2)H]tetradeca-5,8-dienoic acid or a mixture of [2,2 (2)H(2)]- and [8,8,8-(2)H(3)]octanoic acid as supplements. (2)H NMR and mass spectroscopy confirmed the incorporation of (5Z,8Z)-[10,11,12,12-(2)H]tetradeca 5,8-dienoic acid into the C(13) side chain as well as into the C(6) side chain of lipstatin. Moreover, deuterium was incorporated into the C(6) side chain of lipstatin from the 8-position but not from the 2-position of octanoate. The data establish that the beta-lactone moiety of lipstatin is formed by condensation of a C(8) and a C(14) fatty acid with a concomitant exchange of the H-2 atoms of the C(8) fatty acid. PMID- 11421792 TI - Stereoselectivity and regioselectivity in the segment-coupling Prins cyclization. AB - The scope of the segment-coupling Prins cyclization has been investigated. The method is outlined in Scheme 1 and involves esterification of a homoallylic alcohol (1), reductive acetylation to give the alpha-acetoxy ether (3), and cyclization on treatment with a Lewis acid to produce a tetrahydropyran (4). Alkene geometries dictate the product configurations, with E-alkenes leading to equatorial substituents and Z-alkenes leading to axial substituents (Table 1). Not unexpectedly, applying the method to allylic alcohols leads to fragmentation rather than a disfavored 5-endo-trig cyclization. Dienols in which one alkene is allylic and the other alkene is homoallylic cyclize efficiently and produce the tetrahydropyrans 49-54, Table 3. Dienols with two homoallylic alkenes cyclize with modest to high regioselectively, generating tetrahydropyrans 40-45, Table 2. The relative rates for cyclization decrease in the order of vinyl > Z-alkene > E alkene > alkyne. The configurations of the products are consistent with cyclization via a chair conformation, Figure 1. The 2-oxonia Cope rearrangement may be a factor in the regioselectivity of diene cyclizations and in the erosion of stereoselectivity with Z-alkenes. This investigation establishes the stereoselectivity and regioselectivity for a number of synthetically useful segment-coupling Prins cyclizations. PMID- 11421793 TI - New building blocks for fluorinated imidazole derivatives: preparation of beta fluoro- and beta,beta-difluorohistamine. AB - We demonstrate that "FBr" addition to 1-trityl-4-vinyl-1H-imidazole (7) provides a convenient route to side-chain-fluorinated histamines. Thus, addition of "FBr" to the double bond of 7 occurs with Markovnikov regioselectivity to produce 4-(2 bromo-1-fluoroethyl)-1-trityl-1H-imidazole (8). Substitution with azide, reduction, and removal of the trityl group provide beta-fluorohistamine (1) as the dihydrochloride. Elimination of HBr from 8 followed by a second addition of "FBr" gives 4-(2-bromo-1,1-difluoroethyl)-1-trityl-1H-imidazole (15). This was similarly converted to beta,beta-difluorohistamine (2) as the dihydrochloride. PMID- 11421794 TI - Stereoselective synthesis of naturally occurring alpha-methylenebis-gamma butyrolactones: an application of novel oxiranyl "remote" anions. AB - Stereospecific deprotonation of the epoxy proton at the beta-position of the alpha,beta-epoxy esters 5 and 6 yielded oxiranyl "remote" anions 7 and 8, which could then be used for alkylation. The anions 7 and 8 underwent a consecutive aldol lactonization to give, respectively, epoxy lactones 11 and 13 with high stereoselectivity. Generation of the remote anions as well as their stereoselective reactions served as a new synthetic route to the naturally occurring alpha-methylenebis-gamma-butyrolactones, 1. PMID- 11421795 TI - Pi-aromatic and sulfur nucleophilic partners in cationic pi-cyclizations: intramolecular amidoalkylation and thioamidoalkylation cyclization via omega carbinol lactams(1, 2). AB - NaBH(4) reduction of imides 1 and 6a,b,c followed by a pi-cyclization of the resultant N-acyliminium ions, generated in trifluoroacetic acid conditions, afforded two positional isomers, isoindolobenzothiazolinones 4 and 8, respectively. These ring closures proceeded via an intramolecular alpha amidoalkylation with the classical pi-aromatic or the atypical sulfur atom as an internal nucleophile. A ready access to the related six-membered N,S-heterocyclic compounds such as isoindolobenzothiazinones 20a and 21a is also described. During this reaction, we have shown that omega-carbinol lactam precursor 14a led to endocyclic and exocyclic N-acyliminium ions 18a and 19a in equilibrium via the cyclic aza-sulfonium ion A. The latter furnished the expected products 20a and 21a in good yields. Similarly, different omega-carbinol lactams 14b-e substituted at C-angular position afforded the corresponding isoindolobenzothiazinones 20b-e and 21b-e bearing an angular alkyl, aralkyl, or aryl group. In the case of methyl 14b and benzyl 14e groups, an additional amount of the dehydration products 16b and 31 was isolated. These results indicate that the isomerization-pi-cyclization takes place via the cleavage of the thioether linkage in acidic medium. PMID- 11421796 TI - Cyclopentannulation of 3-alkylindoles: a synthesis of a tetracyclic subunit of the kopsane alkaloids. AB - Indoles which bear an alkyl substituent in the 3-position undergo a [3 + 2] annulation reaction when treated with 1,1-cyclopropane diesters in the presence of Yb(OTf)(3) resulting in 2,3-cyclopentanoindolines. Typically, the reactions are performed at elevated temperatures or at ultrahigh pressures. In cases where steric crowding is an issue, ultrahigh pressures are required. In reactions involving substituted cyclopropanes, significant regio- and diastereocontrol was observed. When the substituent was aromatic or olefinic, the reactions took place at ambient temperature and pressure. The applicability of this methodology to the preparation of a key tetracyclic subunit of the kopsane alkaloids was demonstrated. PMID- 11421797 TI - Iridium-catalyzed transfer hydrogenation of alpha,beta-unsaturated and saturated carbonyl compounds with 2-propanol. AB - The selective transfer hydrogenation of alpha,beta-unsaturated carbonyl compounds to saturated ones was achieved by the use of 2-propanol as a hydrogen donor under the influence of catalytic amounts of [Ir(cod)Cl](2), 1,3 bis(diphenylphosphino)propane (dppp), and Cs(2)CO(3). Thus, a variety of conjugated enones were allowed to react with 2-propanol in the presence of the [Ir(cod)Cl](2)/dppp/Cs(2)CO(3) system to give the corresponding saturated carbonyl compounds in good to excellent yields without formation of allylic alcohols. Both dppp and Cs(2)CO(3) were essential components to achieve the reduction satisfactorily. Additionally, the reduction of carbonyl compounds to alcohols was also promoted by the same catalytic system. When the reaction of a 1:1 mixture of a conjugated ketone and a saturated ketone with 2-propanol was carried out in the presence of [Ir(cod)Cl](2) combined with dppp and Cs(2)CO(3), the reduction of the alpha,beta-unsaturated ketone was found to take place in preference to that of the saturated ketone. PMID- 11421798 TI - Olefin epoxidation with bis(trimethylsilyl) peroxide catalyzed by inorganic oxorhenium derivatives. Controlled release of hydrogen peroxide. AB - The replacement of organometallic rhenium species (e.g., CH(3)ReO(3)) by less expensive and more readily available inorganic rhenium oxides (e.g., Re(2)O(7), ReO(3)(OH), and ReO(3)) can be accomplished using bis(trimethylsilyl) peroxide (BTSP) as oxidant in place of aqueous H(2)O(2). Using a catalytic amount of a proton source, controlled release of hydrogen peroxide helps preserve sensitive peroxorhenium species and enables catalytic turnover to take place. Systematic investigation of the oxorhenium catalyst precursors, substrate scope, and effects of various additives on olefin epoxidation with BTSP are reported in this contribution. PMID- 11421799 TI - Lewis-acid catalyzed organic reactions in water. The case of AlCl(3), TiCl(4), and SnCl(4) believed to be unusable in aqueous medium. AB - Classical Lewis acids such as AlCl(3), TiCl(4), and SnCl(4), believed to be unusable as catalysts in aqueous medium, efficiently catalyzed regio- and stereoselective azidolysis and iodolysis of alpha,beta-epoxycarboxylic acids in water at pH 4.0 and 1.5, respectively. The concept of water-tolerant metal-salt is reexamined in direct relationship to the aqua ion hydrolysis constant. PMID- 11421800 TI - 2-Alkylthio-4-oxo-3-quinazolineacetonitriles and analogous thieno[3,2 d]pyrimidineacetonitriles: reaction with thiols via trapped thioimidates. PMID- 11421801 TI - Electronic structure of terpenoids. PMID- 11421802 TI - Photosensitized oxidation of furans. 20.(1) A novel thermal rearrangement of suitably substituted alkoxyfuran endoperoxides via neighboring-group mechanism: synthesis and reactivity of the first functionalized 2-oxetanyl hydroperoxides. PMID- 11421803 TI - Lipase/ruthenium-catalyzed dynamic kinetic resolution of hydroxy acids, diols, and hydroxy aldehydes protected with a bulky group. PMID- 11421804 TI - Thermolysis of a fluorinated indolylfulgide features a novel 1,5-indolyl shift. PMID- 11421806 TI - Location of template on the human ribosome as revealed from data on cross-linking with reactive mRNA analogs. AB - In this review we summarize data on the location of template on the human ribosome that we obtained from cross-linking (affinity labeling) experiments using reactive mRNA analogs. Types of mRNA analogs, model complexes of these analogs with 80S ribosomes, and methods for analysis of the ribosomal components (proteins and rRNA nucleotides) cross-linked with the mRNA analogs are reviewed. From analysis of the cross-linking data, we suggest a scheme for the arrangement of mRNA on the human ribosome and compare the organization of the mRNA binding center on human and Escherichia coli ribosomes. PMID- 11421807 TI - Differences in lipid composition and proliferative activity of rat hepatoma-27 depending on the target organ. AB - Proliferative activity and lipid composition (phospholipids and gangliosides) were studied in rat hepatoma-27 transplanted subcutaneously or intrahepatically (as models for primary and metastasizing tumors). The mitotic index of subcutaneously transplanted hepatoma far exceeded that of the intrahepatically transplanted tumor. The overall amounts of both phospholipids and gangliosides increased appreciably in the subcutaneously growing hepatoma (in contrast to the intrahepatically growing tumor) in comparison to the control hepatic tissue. The ganglioside composition in the tumors differs from that in the liver: ganglioside GD3 appears, whereas gangliosides GD1b and GT1b decrease in amount in the intrahepatic tumor compared to the control liver and disappear in the subcutaneously transplanted hepatoma. In both tumor types, the amounts of both phosphatidylethanolamine and sphingomyelin exceed the control values. Comparison of these results with previously reported data concerning the phospholipid and ganglioside composition in the regenerating rat liver indicates that the difference in the lipid composition between the subcutaneously and intrahepatically growing hepatomas-27 is due to their different proliferative status and also their microenvironment. PMID- 11421808 TI - Peroxidase-catalyzed co-oxidation of 3,3',5,5'-tetramethylbenzidine with 2-amino 4-nitrophenol, 4,4'-dihydroxydiphenylsulfone, and their polydisulfides in aqueous and micellar media. AB - The effects of different concentrations of 2-amino-4-nitrophenol (ANP) and of its polydisulfide (poly(ADSNP)) on peroxidase-catalyzed oxidation of 3,3',5,5' tetramethylbenzidine (TMB) were studied at 20 degrees C in reversed micelles of AOT (0.2 M) in heptane and in mixed reversed micelles of AOT (0.1 M)-Triton X-100 (0.1 M) in isooctane supplemented with 15% hexanol. The oxidation of TMB was activated nearly twofold in the presence of ANP and nearly fourfold in the presence of poly(ADSNP) in reversed micelles of AOT, whereas in the mixed micelles oxidation of the TMB-ANP pair was associated with inhibition of TMB conversion and poly(ADSNP) activated oxidation of TMB. The co-oxidation of TMB with 4,4'-dihydroxydiphenylsulfone (DDS) and with its polydisulfide (poly(DSDDS)) at different concentrations of phenol components was accompanied by activation of TMB conversion in 0.01 M phosphate buffer (pH 6.4) supplemented with 5% DMF and in reversed micelles of AOT in heptane. The effect of pH of the aqueous solution on the initial oxidation rate of the TMB-DDS and TMB-poly(DSDDS) pairs and also the effect of hydration degree of reversed micelles of AOT on conversion of the same pairs by peroxidase were studied. A scheme of peroxidase-dependent co oxidation of "aromatic amine-phenol" pairs is proposed and discussed. A significant part of this scheme is a nonenzymatic exchange of phenoxyl radicals with amines and of aminyl radicals with phenols. PMID- 11421809 TI - Isolation and study of some properties of laccase from the basidiomycetes Cerrena maxima. AB - A new strain producing extracellular laccase (Cerrena maxima 0275) was found by screening of isolates of Basidiomycetes, and the dynamics of laccase biosynthesis by this strain was studied. The enzyme was purified to homogeneity. The molecular weight of the enzyme is 57 kD, and its pI is 3.5. The activity is constant at pH values in the range 3.0-5.0. The temperature optimum for activity is 50 degrees C. The thermal stability of the laccase was studied. The catalytic and Michaelis constants for catechol, hydroquinone, sinapinic acid, and K4Fe(CN)6 were determined. The standard redox potential of type 1 copper in the enzyme is 750 +/ 5 mV. Thus, the investigated laccase is a high redox potential laccase. PMID- 11421810 TI - Effect of two conserved amino acid residues on DREB1A function. AB - Transcription factors of the DREBP subgroup and the EREBP subgroup contain conserved DNA-binding domains called AP2/EREBP domains, which specifically bind to DRE cis-element and GCC-box, respectively. The 14th and 19th amino acid residues of AP2/EREBP domains are absolutely conserved in the transcription factors of the DREBP subgroup as well as in the EREBP subgroup. However, these two residues of transcription factors of the DREBP subgroup are different from those of the EREBP subgroup. To assess the functional significance of these two residues in binding to the target sequence, the Val (14th residue) and Glu (19th residue) of the AP2/EREBP domain of DREB1A (a transcription factor of the DREBP subgroup) were mutated individually or doubly to Ala and Asp, respectively. This made the 14th and 19th amino acid residues of mutant DREB1A identical to the corresponding residues of transcription factors of the EREBP subgroup. Yeast in vivo analysis showed that: 1) on a selective medium plate of SD/His- Ura- Trp- + 30 mM approximately 60 mM 3-AT, the growth of yeast cells containing HIS and lacZ double reporter genes was normal in the transformation of the 19th singly mutated DREB1A, obviously inhibited in the transformation of the 14th singly mutated DREB1A, and seriously inhibited in the transformation of the 14th/19th doubly mutated DREB1A; 2) quantitative assay of beta-galactosidase activity showed that the intensities of lacZ expression decreased in the transformations of the 14th singly mutated and 14th/19th doubly mutated types. The experimental results revealed that the 19th site mutation did not affect the binding of the DREB1A transcription factor to the DRE cis-element; the 14th site mutation obviously inhibited their binding; and the double mutation of the 14th/19th sites seriously inhibited their binding. This suggests that the conserved Val (14th) and Glu (19th) residues are crucial in the regulation of the binding activity of DREB1A to the DRE cis-element. PMID- 11421811 TI - Investigation of calcium accumulation in mitochondria in cells undergoing apoptosis. AB - One of the earliest features of apoptosis is the induction of the mitochondrial permeability transition (MPT) due to opening of a pore in the mitochondrial membrane. We estimated the Ca2+ capacity of mitochondria (a threshold level of Ca2+ that induces the release of this cation from mitochondria) during apoptosis. Incubation of thymocytes at 37 degrees C for 4 h equally decreased the mitochondrial Ca2+ capacity both in the presence and the absence of dexamethasone, an inducer of apoptosis. At the same time, dexamethasone significantly stimulated internucleosomal DNA fragmentation, which is one of the manifestations of apoptosis. Cyclosporin A prevented the time-dependent decrease in the Ca2+ capacity of mitochondria but did not affect internucleosomal DNA fragmentation. Therefore, induction of apoptosis assessed by internucleosomal DNA fragmentation is not mediated by the mitochondrial permeability transition. PMID- 11421812 TI - Regulation of free radical processes by delta-sleep inducing peptide in rat tissues under cold stress. AB - An intraperitoneal injection of an exogenous delta-sleep inducing peptide (DSIP) at a dose of 12 microg/100 g body weight shifted the prooxidant-antioxidant balance of free radical process (FRP) in tissues and erythrocytes of rats: the activities of antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase) and the concentrations of antioxidants (reduced glutathione in particular) increased. The DSIP stimulated the myeloperoxidase activity in blood neutrophils and had no effect on the activity of xanthine oxidase, a prooxidant enzyme, in the brain and liver. Cold stress displaced the prooxidant-antioxidant balance by increasing the xanthine oxidase activity in tissues and decreasing the myeloperoxidase activity in blood neutrophils; it also inhibited the enzyme antioxidant activities in tissues and erythrocytes that was neutralized by an increased ceruloplasmin activity in blood plasma and by an elevated level of antioxidants in rat blood and tissues. Preliminary administration of DSIP to animals exposed to cold stress restored the prooxidant-antioxidant balance: it normalized the myeloperoxidase activity in blood neutrophils, decreased the xanthine oxidase activity, and increased the activity of antioxidant enzymes in tissues and erythrocytes restoring the antioxidant level. The molecular regulation mechanism of free radical processes by DSIP in tissues under stressful conditions is discussed. PMID- 11421813 TI - Hydrogen peroxide inhibits photosynthetic electron transport in cells of cyanobacteria. AB - The effect of H2O2 on photosynthetic O2 evolution and photosynthetic electron transfer in cells of cyanobacteria Anabaena variabilis and Anacystis nidulans was studied. The following experiments were performed: 1) directly testing the effect of exogenous H2O2; 2) testing the effect of intracellular H2O2 generated with the use of methyl viologen (MV); 3) testing the effect of inhibiting intracellular H2O2 decomposition by salicylic acid (SA) and 3-amino-1,2,4-triazole (AT). H2O2 inhibited photosynthetic O2 evolution and light-induced reduction of p benzoquinone (BQ) + ferricyanide (FeCy) in the Hill reaction. The I50 value for H2O2 was ~0.75 mM. Photosynthetic electron transfer in the cells treated with H2O2 was not maintained by H2O2, NH2OH, 1,5-diphenylcarbazide, tetraphenylboron, or butylated hydroxytoluene added as artificial electron donors for Photosystem (PS) II. The H2O --> CO2, H2O --> MV (involving PSII and PSI) and H2O --> BQ + FeCy (chiefly dependent on PSII) electron transfer reactions were inhibited upon incubation of the cells with MV, SA, or AT. The N,N,N,N-tetramethyl-p phenylenediamine --> MV (chiefly dependent on PSI) electron transfer was inhibited by SA and AT but was resistant to MV. The results show that H2O2 inhibits photosynthetic electron transfer. It is unlikely that H2O2 could be a physiological electron donor in oxygenic photosynthesis. PMID- 11421814 TI - Isolation, purification, and characterization of catalase from the methylotrophic yeast Pichia pastoris. AB - Catalase (CATpp) with molecular weight 223 kD was isolated from the methylotrophic yeast Pichia pastoris and purified 90-fold by ion-exchange chromatography and gel filtration. Quantitative parameters of absorption and CD spectra of CATpp solutions and of its membrane-concentrated form (CATpp-conc) were studied. Rates of H2O2 decomposition and kinetic characteristics Km and kcat of CATpp and CATpp-conc were determined in 10 mM phosphate buffer (pH 7.4) at 30 degrees C, as well as the effective constant kin of the enzyme inactivation rate during the catalysis and the constant k2 of the interaction rate of the Complex I catalases with H2O2. Thermal inactivation of CATpp in solutions at 45 degrees C was characterized by the effective rate constant kin*, and the low-frequency (27 kHz) ultrasonic inactivation of CATpp at 20 degrees C was characterized by the first-order rate constant kin(US). All spectral and kinetic characteristics of CATpp and CATpp-conc were compared with the corresponding values for catalase from bovine liver (CAT) and for catalase from the methylotrophic yeast Candida boidinii (CATcb). All three catalases were rather similar in their spectral properties but strongly varied in their kinetic parameters, and their comparison suggests that CATpp should be the best enzyme in its overall properties as it displayed the maximal efficiency in terms of kcat/Km, thermal stability comparable with the thermal stability of CAT in terms of kin*, the minimal kin, and high stability in the ultrasonic cavitation field at the US power of 60 W/cm2. PMID- 11421815 TI - Inhibition of human platelet aggregation by amides and ester of salicylic acid with platelet-activating factor analogs. AB - The influence of acetyl salicylic acid (ASA) derivatives with platelet-activating factor (PAF) lipid analogs on PAF-induced human platelet aggregation has been studied. It was found that the ASA amide with an ethanolamine plasmalogen PAF analog (1-0-alk-1'-enyl-2-acetyl-sn-glycero-3-phospho-(N-2' acetoxybenzoyl)ethanolamine) and the ASA ester with a choline plasmalogen PAF analog (1-0-alk-1'-enyl-2-(2'-acetoxybenzoyl)-sn-glycero-3-phosphocholine) at concentrations of 10-7-10-6 M effectively inhibit PAF-induced aggregation of human platelets. In contrast to these compounds, the ASA amide with an alkyl PAF analog (1-0-alkyl-2-acetyl-sn-glycero-3-phospho-(N-2' acetoxybenzoyl)ethanolamine) did not inhibit PAF-induced platelet aggregation. As possible mechanisms of action of the studied compounds, the blockade of PAF receptor and cyclooxygenase inhibition are proposed. PMID- 11421816 TI - Influence of acidic exopolysaccharide of Xanthomonas campestris IBPM 124 on the kinetic parameters of extracellular bacteriolytic enzymes. AB - Interactions of a negatively charged exopolysaccharide of Xanthomonas campestris IBPM 124 with its extracellular enzymes (muramidase, endopeptidase, and neutral phosphatase) and also with egg lysozyme, lysostaphin, muramidase of Streptomyces globisporus, and a bacteriolytic enzyme complex of Streptomyces albus were studied. All these enzymes were positively charged under the conditions of their maximal activity. It was shown that interaction of the acidic exopolysaccharide from X. campestris with these enzymes changed their kinetic parameters. The change was either positive (increase in reaction rate) or negative (decrease in reaction rate) and depended on the enzyme and type of substrate cleaved. Due to such interactions, the acidic exopolysaccharide secreted by X. campestris into the environment not only retained and transported positively charged exoenzymes into the near-cellular space, but also regulated their activity. PMID- 11421817 TI - Site-directed mutagenesis of cytochrome b5 for studies of its interaction with cytochrome P450. AB - We have shown earlier that microsomal cytochrome b5 can form a specific complex with mitochondrial cytochrome P450 (cytochrome P450scc). The formation of the complex between these two heme proteins was proved spectrophotometrically, by affinity chromatography on immobilized cytochrome b5, and by measuring the cholesterol side-chain cleavage activity of cytochrome P450scc in a reconstituted system in the presence of cytochrome b5. To further study the mechanism of interaction of these heme proteins and evaluate the role of negatively charged amino acid residues Glu42, Glu48, and Asp65 of cytochrome b5, which are located at the site responsible for interaction with electron transfer partners, we used site-directed mutagenesis to replace residues Glu42 and Glu48 with lysine and residue Asp65 with alanine. The resulting mutant forms of cytochrome b5 were expressed in E. coli, and full-length and truncated forms (shortened from the C terminal sequence due to cleavage of 40 amino acid residues) of these cytochrome b5 mutants were purified. Addition of the truncated forms of cytochrome b5 (which do not contain the hydrophobic C-terminal sequence responsible for interaction with the membrane) to the reconstituted system containing cytochrome P450scc caused practically no stimulation of catalytic activity, indicating an important role of the hydrophobic fragment of cytochrome b5 in its interaction with cytochrome P450scc. However, full-length cytochrome b5 and the full-length Glu48Lys and Asp65Ala mutant forms of cytochrome b5 stimulated the cholesterol side-chain cleavage reaction catalyzed by cytochrome P450scc by 100%, suggesting that residues Glu48 and Asp65 of cytochrome b5 are not directly involved in its interaction with cytochrome P450scc. The replacement of Glu42 for lysine, however, made the Glu42Lys mutant form of cytochrome b5 about 40% less effective in stimulation of the cholesterol side-chain cleavage activity of cytochrome P450scc, indicating that residue Glu42 of cytochrome b5 is involved in electrostatic interactions with cytochrome P450scc. Residues Glu42 and Glu48 of cytochrome b5 appear to participate in electrostatic interaction with microsomal type cytochrome P450. PMID- 11421818 TI - Interaction between duodenase and alpha1-proteinase inhibitor. AB - The interaction between duodenase, a newly recognized serine proteinase belonging to the small group of Janus-faced proteinases, and alpha1-proteinase inhibitor (alpha1-PI) from human serum was investigated. The stoichiometry of the inhibition was 1.2 mol/mol. The presence of a stable enzyme-inhibitor complex was shown by SDS-PAGE. The mechanism of interaction between duodenase and alpha1-PI was shown to be of the suicide type. The equilibrium and inhibition constants are 13 +/- 3 nM and (1.9 +/- 0.3).105 M-1.sec-1, respectively. Based on the association rate constant of the enzyme-inhibitor complex and localization of duodenase and alpha1-PI in identical compartments, alpha1-PI is suggested to be a duodenase inhibitor in vivo. PMID- 11421819 TI - 16alpha,17alpha-cycloalkane derivatives of progesterone intensively bind to a rat serum protein. AB - The interaction of 6alpha-methyl-[1,2-3H]16alpha,17alpha-cyclohexanoprogesterone with rat serum proteins has been studied. Specific binding of this ligand characterized by Kd = 0.36 +/- 0.10 microM and concentration of binding sites (Bmax) of about 1 microM (27.8 +/- 12.5 pmol/mg total protein) was found. According to competitive analysis, the affinity of the studied progestins to a protein that differs from transcortin was to some extent correlated with their hydrophobicity. The dissociation kinetics of 3H-ligand-protein complexes were biphasic, the binding sites forming stable and labile complexes with 3H-ligand being eluted in the same region during ion-exchange chromatography. In overall properties, the serum protein differs from the progesterone receptor and the pregna-D'-pentarane-specific protein from rat uterus. It is suggested that the revealed protein may provide high progestagenic activity of 6alpha-methyl 16alpha,17alpha-cyclohexanoprogesterone by prolonging its retention in the bloodstream. PMID- 11421820 TI - Properties of bacteriophage T4 baseplate protein encoded by gene 8. AB - Gene product 8 (gp8, 344 amino acids per monomer) of bacteriophage T4 is one of the baseplate structural proteins. We constructed an expression vector of gp8 and developed a method for purification of recombinant protein. CD spectroscopy showed that gp8 is an alpha/beta type structural protein. Its polypeptide chain consists of nearly 40% beta-structure and 15% alpha-helix. These data agree with results of prediction of secondary structure based on the amino acid sequence of the protein. The sedimentation coefficient under standard conditions (S20,w) is 4.6S. Analytical ultracentrifugation results demonstrated that gp8 in solution has two types of oligomers--dimer and tetramer. The tetramer of gp8 may be included in the wedge (1/6 of the baseplate), and the dimer may be an intermediate product of association. PMID- 11421821 TI - Meta-analysis - a valuable but easily misused tool. PMID- 11421822 TI - The influence of a dominating centre on a quantitative systematic review of granisetron for preventing postoperative nausea and vomiting. AB - BACKGROUND: We performed a meta-analysis on granisetron in the prevention of postoperative nausea and vomiting (PONV) and further investigated whether total results and the dose-response characteristics may be significantly affected by a single centre. METHODS: Systematically searched randomised controlled trials (RCT) using granisetron for the prevention of PONV after general anaesthesia were included in the analysis. The pooled relative risks (RR) and numbers needed to treat (NNT) with their corresponding 95%-confidence intervals (CI) were calculated. For all centres, one dominating centre and other centres pooled, comparisons were performed according to all doses, low dose (20 microg/kg) granisetron. RESULTS: A total of 27 RCT with 2938 patients were included in the analysis. RR (CI) to suffer from PONV with granisetron when all comparisons were considered was 0.46 (0.39-0.54), 0.7 (0.6 0.81) and 0.34 (0.28-0.41) for all doses, low and high dose, respectively. RR of the dominating centre (1867 patients) were significantly better compared to the remaining centres (1071 patients), with 0.41 (0.34-0.49) and 0.60 (0.49-0.73), respectively. In the dominating centre low dose granisetron was ineffective with a RR of 0.84 (0.68-1.04), while high dose granisetron led to a strong decrease with a RR of 0.30 (0.26-0.36). In contrast, the RR of other centres pooled for low and high dose granisetron were comparable with 0.62 (0.49-0.79) and 0.56 (0.42-0.75), respectively. CONCLUSIONS: Overall results and dose-response characteristics of meta-analyses may be significantly altered by one dominating centre. Further, if data of a dominating centre do not appear to be valid for other centres, it may seem advisable to either exclude them from the analysis or to perform sub-group analyses so that results without the data from the dominating centre are available. PMID- 11421823 TI - The systemic inflammatory response after cardiac surgery with cardiopulmonary bypass in children. AB - Paediatric cardiac surgery often requires cardiopulmonary bypass (CPB) during the surgical intervention. CPB is known to elicit a systemic inflammatory response with activation of the complement and coagulation systems, stimulation of cytokine production, cellular entrapment in organs, neutrophil activation with degranulation, platelet activation, and endothelial dysfunction. These changes are associated with a risk of postoperative organ dysfunction and increased morbidity and mortality in the postoperative period. Clinical studies have concentrated on measurement of inflammatory markers and mediators in peripheral blood, where the systemic inflammatory response in the paediatric cardiac patient seems to be different from the adult case. Looking at the organ level, experimental studies have the advantage of providing information contributing to a better understanding of the pathological events that may lead to the deteriorated organ function. This review focuses on the systemic inflammatory response after cardiac surgery with CPB in children and experimental CPB models. PMID- 11421824 TI - Lack of agreement between thermodilution and carbon dioxide-rebreathing cardiac output. AB - BACKGROUND: A continuous, accurate, non-invasive monitor of cardiac output would represent a major step forward in patient management. A cardiac output computer, NICO2, based on the Fick principle and an automatic partial carbon dioxide (CO2) rebreathing technique has just become available. We compared the performance of this monitor with the standard thermodilution method. METHODS: Thirty patients were investigated after cardiac surgery. Replicate measurements were performed simultaneously with the thermodilution and NICO2 techniques. An Altman-Bland analysis was used to assess repeatability of each of the two methods and to determine the agreement between the two techniques. RESULTS: The repeatabilities of thermodilution and CO2-rebreathing cardiac output were excellent, with coefficients of repeatability of 0.35 l/min and 0.60 l/min. Mean thermodilution and NICO2 cardiac output were 4.4 l/min (SD 0.9, range 2.7-6.1) and 4.6 l/min (SD 1.3, range 1.6-6.9). A comparison of the methods, however, revealed excessive limits of agreement (+/-1.80 l/min). CONCLUSION: The agreement between the NICO2 derived cardiac output and the de facto standard - thermodilution cardiac output is poor. The methods are not interchangeable with the present version of the NICO2. The repeatability of the partial CO2-rebreathing technique holds promise that a sufficient accuracy may be obtained by suitable modifications of the monitor's algorithms. PMID- 11421825 TI - High incidence of pruritus after large doses of hydroxyethyl starch (HES) infusions. AB - BACKGROUND: There are several studies indicating a correlation between treatment with hydroxyethyl starch (HES) and pruritus. In order to see whether there is a possible dose-response relationship between HES and pruritus, we retrospectively studied 50 patients who had received HES in varying doses (cumulative dose 500 19500 ml) as hemodilution therapy after subarachnoid hemorrhage. METHODS: Of 50 consecutive patients, 6 were excluded due to severe neurological sequelae. A questionnaire was sent to the remaining 44 patients at 6 months (5-12 months) median (range) after the end of HES treatment. RESULTS: We received answers from 37 patients, of whom 54% reported pruritus. On average pruritus lasted for 15 weeks. There was significantly more pruritus in patients who received more than 5000 ml of HES versus those who received less than 5000 ml (P=0.023). Pruritus had a delayed onset and appeared as pruritic crises lasting for 2-30 min. It had a patchy distribution in most patients and no predilected locations. In 4 patients (20%) the pruritus lasted longer than 21 weeks. CONCLUSION: Our study indicates that there is a dose-dependency for the incidence of HES-induced pruritus, and that in some cases the pruritus may be severe and long-lasting. PMID- 11421826 TI - The pressure at the lower inflexion point has no relation to airway collapse in surfactant-treated premature lambs. AB - BACKGROUND: The lower inflexion point (LIP) on the inspiratory part of the pressure-volume (PV) loop has been suggested to be related to the pressure at which air spaces collapse. Our hypothesis is that airway collapse might instead be assessed from the upper inflexion point on the expiratory part of the PV-loop (UIPexp), where lung volume starts to decrease significantly. We therefore examined whether there was a relation between LIP and UIPexp in premature surfactant-treated lambs. METHODS: Ten lambs, at 119-141 days of gestational age, were delivered by cesarean section and given 200 mg/kg modified natural porcine surfactant before the first breath. The lambs were then connected to a ventilator and PV-loops using airway pressures of 0-35-0 (ZEEP-loop) and 5-35-5 cmH2O (PEEP loop) were obtained after lung recruitment at 15, 60 and 120 min after birth. From the loops, LIP, UIPexp, upper inflexion point of the inspiratory part of the loop (UIP insp), inspiratory capacity (IC) as well as inspiratory and expiratory maximal compliance of the respiratory system (Crs(insp) and Crs(exp)) were calculated. RESULTS: The ZEEP-loop showed a substantial hysteresis with a distinct LIP at 19+/-2 cmH2O (mean+/-SD), which was different (P<0.001) from UIPexp (9+/-2 cmH2O). The pressures at LIP and UIPexp were unrelated (r2=0.06). UIPinsp was located at 28+/-2 cmH2O. Crs(insp) was 2.1+/-0.6 ml x cmH2O(-1) x kg( 1), which was lower (P<0.001) than Crs(exp) (2.8+/-0.6 ml x cmH2O(-1) x kg(-1)). IC was 26+/-6 ml/kg. The PEEP-loop had a minimal hysteresis with an expiratory part coinciding with that of the ZEEP-loop. CONCLUSION: In surfactant-treated premature lambs the pressures at LIP and UIPexp are not related, showing that LIP does not indicate the pressure at which airways collapse. PMID- 11421827 TI - Dynamic changes in cerebral oxygenation related to deep hypothermia and circulatory arrest evaluated by near-infrared spectroscopy. AB - BACKGROUND: Total circulatory arrest in deep hypothermia, which is used in corrective surgery of complex cardiovascular malformations, has been said to cause brain injury. Near-infrared spectroscopy (NIRS) is a new non-invasive method that potentially monitors changes in cerebral oxygenation and tissue oxygen utilisation. The aim of this experimental study in rabbits was to evaluate the change in intravascular and intracellular oxygenation patterns during cooling, deep hypothermic circulatory arrest and rewarming using a commercially available NIRS-cerebral oximeter. METHODS: Ten New Zealand White male rabbits (weight, 3.1+/-0.25 kg BW) were included in this study. All animals underwent cardiopulmonary bypass (CPB), cooling to a rectal temperature below 15 degrees C, 60 min of deep hypothermic circulatory arrest (DHCA) followed by reperfusion and rewarming. Cerebral oxyhaemoglobin (HbO2), deoxyhaemoglobin (HHb) and cytochrome oxidase aa3 (CytOxaa3) concentrations were continuously measured during the entire procedure using the Cerebral RedOx Monitor 2020 (Criticon cerebral redox monitor 2020, Johnson & Johnson Medical). Total haemoglobin concentration (tHb) and regional cerebral oxygen saturation (rSO2) values were calculated by integrated algorithm. RESULTS: In all animals an initial increase of oxygenated haemoglobin (HbO2, rSO2) and a fall in deoxygenated haemoglobin (HHb) were found during cooling on bypass. A slight decrease in CytOxaa3 signal was observed in response to initial cooling. Variation in intravascular haemoglobin oxygenation parameters (HbO2, HHb) were related to haemodynamic changes associated with fluid loading, initiation and termination of CPB, bypass flow rate and cooling and rewarming. When the pump flow was stopped all NIRS parameters, except the HHb value, decreased precipitously during the DHCA-period (P<0.01). After reperfusion and rewarming, all haemoglobin oxygen saturation readings returned nearly to pre CPB levels (P=0.09), but the CytOxaa3 was still significantly lower than the pre CPB levels (P<0.05). CONCLUSION: The change in the NIRS-derived haemoglobin oxygenation parameters may reflect physiological changes in systemic and cerebral haemodynamics. CytOxaa3 values may represent related effects on cellular oxygenation. Thus, continuous, real-time NIRS-monitoring may identify critical periods with inadequate brain tissue oxygenation, particularly during DHCA. The neurological implications of the observed changes in NIRS oxygenation parameters, however, require further quantitative morphological evaluation of the brain in animals surviving a longer reperfusion and observation period. PMID- 11421828 TI - Heart-lung interactions during positive pressure ventilation: left ventricular pressure-volume momentary response to airway pressure elevation. AB - BACKGROUND: Left ventricular (LV) pressure and volume changes are known to occur in response to positive airway pressure (PAP). We aimed to further describe the immediate LV response to increased PAP as demonstrated in successive heart cycles with LV pressure and volume alterations. We postulated that these acute systematic LV events during institution of PAP can follow a distinct pattern that would allow calculation of parameters of systolic function, including end systolic elastance (Ees) and preload recruitable stroke work (PRSW). We also aimed to examine the relationship of PAP-derived Ees and PRSW to the same parameters derived from vascular occlusion. METHODS: Eight anesthetized adult pigs were studied with invasive circulatory measurements including LV pressure and volume (conductance). The PAP intervention was an airway pressure plateau of 15 cm H2O for 6 s (APP). Venous occlusion was performed by transient balloon inflation in the inferior vena cava (IVCO). Ees and PRSW were derived for each APP and IVCO intervention. RESULTS: Central circulatory variables during APP and IVCO are reported. LV systolic function parameters could be derived from each of the heart-lung interactions during APP sequences. Ees and PRSW derived from APP showed a significant positive bias in relation to those derived from the IVCO sequence. CONCLUSIONS: We conclude that the heart-lung interactions during APP of the magnitude and duration shown here can allow derivation of Ees and PRSW. These parameters are not interchangeable with Ees and PRSW derived from IVCO. PMID- 11421829 TI - Risk factors for nosocomial intensive care infection: a long-term prospective analysis. AB - BACKGROUND: To identify risk factors for nosocomial infection in intensive care and to provide a basis for allocation of resources. METHODS: Long-term prospective incidence study of risk factors for nosocomial infection in the surgical-medical intensive care unit of a university hospital. RESULTS: A total of 2671 patients were admitted during four years, and 562 of 574 patients staying >48 h were observed during 4921 patient days (median length of stay 5 days, range 2-114). Of these, 196 (34%) patients had 364 nosocomial infections after median 8 10 days, an infection rate of 14/100 admissions. Infection prolonged length of stay 8-9 days and doubled the risk of death. The infections were 17% blood stream, 26% pneumonias, 34% wound, 10% urinary tract and 13% other infections. The incidence of bloodstream infection declined significantly during the study years, from 12% to 5%. In multiple regression analysis, the important variables for infection were central venous catheter, mechanical ventilation, pleural drainage and trauma with open fractures. High age, immunosuppression and infection on admission did not influence the risk of acquiring infection. Trauma patients constituted 24% of the study population. Trauma with open fractures increased the risk of infection more than twice (P=0.003), mainly due to wound infections. CONCLUSION: Trauma cases, with open fractures, were the patients most at risk of infection, despite low disease severity scores. Resources to prevent nosocomial infection should be allocated to these patients. PMID- 11421830 TI - Studies on fluid extravasation related to induced hypothermia during cardiopulmonary bypass in piglets. AB - BACKGROUND: Hypothermia, commonly used for organ protection during cardiopulmonary bypass (CPB), has been associated with changes in plasma volume, hemoconcentration and microvascular fluid shifts. Fluid pathophysiology secondary to hypothermia and the mechanisms behind these changes are still largely unknown. In a recent study we found increased fluid needs during hypothermic compared to normothermic CPB. The aim of the present study was to characterize the distribution of the fluid given to maintain normovolemia. In addition, we wanted to investigate the quantity and quality of the fluid extravasated during hypothermic compared to normothermic CPB. METHODS: Two groups of anesthetized piglets were studied during 2 h of hypothermic (28 degrees C) (n=7) or normothermic (38 degrees C) (n=7) CPB. Net fluid balance (input-output) was recorded. Changes in colloid osmotic pressures of plasma (COPp) and interstitial fluid (COPi), plasma volume (PV), hemoglobin (Hb), hematocrit (HCT), mean corpuscular volume (MCV), s-osmolality, s-albumin and s-total protein was followed throughout the experiments. Fluid extravasation rate was calculated. In addition, total tissue water content was measured and compared with a control group (n=6) (no CPB). RESULTS: During hypothermic compared with normothermic CPB, the average net positive fluid balance from 10-120 min of extracorporeal circulation was 1.35+/-0.06 ml x kg(-1) x min(-1) and 0.33+/-0.03 ml x kg(-1) x min(-1) respectively (P<0.0001). We found a marked increase in fluid extravasation during hypothermic CPB. The extravasation rate during hypothermia was 1.8+/-0.2 ml x kg(-1) x min(-1), (1st hour) and 1.1+/-0.2 ml x kg(-1) x min( 1) (2nd hour) compared with 0.8+/-0.2 ml x kg(-1) x min(-1), and 0.1+/-(0.1) ml x kg(-1) x min(-1) during normothermia, respectively (P<0.01). The total intravascular protein and albumin masses remained constant in both groups. Following hypothermic CPB, the water content increased significantly in all tissues and organs. CONCLUSION: During hypothermic CPB an increased extravasation of fluid from the intravascular to the interstitial space was found. As no leakage of proteins could be demonstrated, based on stable values for albumin and protein masses throughout the experiments, the extravasated fluid contained mainly water and small solutes. PMID- 11421831 TI - Always flush the sampling port before flushing the arterial cannula in pediatric patients. AB - BACKGROUND: Blood sampling from arterial lines is a frequent event in anesthesia and critical care. To avoid clot formation, both the stopcock outlet and the cannula must be flushed after sampling. We investigated in a bench experiment whether fluid flow through the cannula is affected by the sequence of flushing procedures. METHODS: Continuity of fluid delivery from a vascular cannula was gravimetrically determined using two different flushing techniques with either a syringe pump flush system or a bag flush system. The procedures comprised first flushing the stopcock towards the cannula and then towards the stopcock sampling outlet or the reverse order. Experiments were repeated in triplicate and two sets for each flushing system at hydrostatic pressures of 37 mm Hg and 74 mm Hg. RESULTS: The main finding of the study was that flushing the stopcock towards the outlet after flushing the cannula resulted in considerable retrograde aspiration volumes and zero flow times, in particular in combination with syringe pump flush systems. At a hydrostatic pressure of 74 mm Hg, the observed zero flow time at the cannula tip amounted to (mean+/-SD) 0.1+/-0.01 min with the bag flush system and 7.7+/-0.5 min with the syringe pump flush system. The related retrograde aspiration volumes were 2.2+/-0.7 microl with the bag system and 30.0+/-2.0 microl with the syringe pump system. No backflow was recorded when the stopcock was first flushed to ambient pressure and then afterwards towards the cannula. CONCLUSION: Opening a flush system to ambient pressure affects the continuity of fluid delivery, particularly when using syringe pump flush systems. After blood sampling, the stopcock outlet should be flushed first followed by cannula flushing. PMID- 11421832 TI - Gentamicin dosing in critically ill patients. AB - Gentamicin is used worldwide in the treatment of serious infections in critically ill patients. The therapeutic efficacy of gentamicin is correlated to the peak serum concentration and the adverse effects to the trough concentrations. Information concerning the pharmacodynamics in critically ill patients is scarce, but pharmacokinetic data are available. A once-daily dosage regimen has replaced multiple dosing of gentamicin in most intensive care units. No studies evaluating the superiority of either of these dosage recommendations in critically ill patients have ever been conducted. Based on 8 meta-analyses performed addressing this issue on a wide range of patients and theoretical considerations, we consider a once-daily dosage regimen feasible in critically ill patients. In septic patients the volume of distribution is significantly increased compared to normal patients, implying that the initial dose should be increased in this patient population. Additionally a general trend towards using higher loading doses (5-7 mg/kg) has been observed in USA, and the appropriateness of this dosing strategy is based on a large descriptive American study. We recommend that the initial dosage of gentamicin in critically ill hyperdynamic septic patients should be 7 mg/kg. Optimal and appropriate monitoring of the treatment with gentamicin in the critically ill patient is still an issue for further investigation. The treatment period with gentamicin should be short (3-5 days), bearing the pharmacological properties of aminoglycosides (small volume of distribution and poor tissue penetration) in mind. In patients with reduced renal function the initial dose of gentamicin should also be increased and maintenance dose reduced preferentially by prolonging the dosing intervals. However, the use of aminoglycosides in a high dose regimen in oliguric or anuric patients or patients who present with a rapidly decreasing renal function needs further consideration. PMID- 11421833 TI - Effects of prilocaine and articaine on human leucocytes and reactive oxygen species in vitro. AB - BACKGROUND: The aim of this study was to investigate the ability of local anaesthetics to inhibit reactive oxygen and nitrogen species generated by either stimulated human leucocytes or cell-free systems using luminol chemiluminescence (CL). METHODS: Free radical generation was stimulated in leucocyte assay by formyl-methionyl-leucyl-phenylalanine (FMLP, 2 microM). In cell-free experiments, hydrogen peroxide (H2O2) 3.5 mM, sodium hypochloride 5 microM, ferrous sulphate (FeSO4) 40 nM, peroxynitrite 50 nM and xanthine 0.1 mmol l(-1) plus xanthine oxidase 0.25 U ml(-1) were used to produce H2O2, hypochlorous acid (HOCl), hydroxyl radical, peroxynitrite and superoxide-induced CL, respectively. RESULTS: Prilocaine inhibited FMLP-induced CL in leucocytes (94+/-1%, at 1 mM), whereas articaine showed an activation (59+/-7%) at high concentration (1 mM) and inhibition (13+/-6%) at low concentration (0.1 mM). In cell-free experiments, prilocaine (22+/-6%, at 1 mM) and articaine (85+/-1%, at 1 mM) caused concentration-dependent inhibition in xanthine-xanthine oxidase-induced CL. Although articaine had no effect on H2O2-induced CL, prilocaine significantly attenuated the H2O2 signal (97+/-0.3%, at 1 mM). Prilocaine (99+/-0.04%, 1 mM) and articaine (70+/-6%, 1 mM) markedly inhibited HOCl-induced CL, whereas these drugs had no effect on FeSO4-induced CL. Articaine inhibited peroxynitrite CL (63+/-6%, 1 mM), but prilocaine did not produce any depression on this signal. CONCLUSION: Prilocaine interacted with superoxide, HOCl and H2O2, whereas articaine reacted with superoxide, HOCl, and peroxynitrite. The direct scavenging properties of these drugs might be involved in the inhibition observed in leucocyte assay and could provide experimental support for investigating the potential benefit of using these local anaesthetics in patients presenting pathologies associated with free radical reactions. PMID- 11421834 TI - Frequency of haemoglobin desaturation with the use of succinylcholine during rapid sequence induction of anaesthesia. AB - BACKGROUND: The perceived safety of the use of succinylcholine is based on the fact that recovery from its effects will occur before oxygen desaturation occurs in case of failure to intubate or ventilate. The purpose of this study was to examine the incidence of oxygen desaturation after the use of succinylcholine prior to resumption of spontaneous ventilation following four different preoxygenation techniques. METHODS: Twenty-five patients each were randomly allocated to preoxygenation with 4 deep breaths of 100% oxygen or by breathing oxygen for 1, 3 or 5 min following which they received a rapid sequence induction of anaesthesia with fentanyl 1 microg kg(-1), a sleep dose of thiopentone and succinylcholine 1 mg kg(-1). Oxygen saturation was monitored continuously using a finger probe. Ventilation was not assisted unless the saturation decreased to 60 s). The estimated degree of difficulty was not significantly different. CONCLUSION: In conclusion, both video-assisted intubation devices, the video-optical intubation stylet and the angled video intubation laryngoscope, can be considered to be simple and effective tools which facilitate tracheal intubation and which require only brief instruction and training. PMID- 11421838 TI - Ultrasound control for presumed difficult epidural puncture. AB - BACKGROUND: The efficacy of epidural anaesthesia depends on the accurate identification of the epidural space (ES). Abnormal anatomical conditions may make the procedure difficult or impossible. The aim of this study was to investigate whether pre-puncture ultrasound examination of the spinal anatomy might be beneficial in expected cases of difficult epidural anaesthesia. METHODS: We used digital ultrasound equipment with a 5-MHz transducer to assess the anatomy of the ES and the posterior parts of the spinal column. We examined 72 parturients with abnormal anatomical conditions who were scheduled for epidural anaesthesia. The women were randomised into two equal groups. In all patients, the standard loss of resistance technique was used. In the ultrasound group, an ultrasound examination of the appropriate spinal region was conducted prior to epidural puncture. ES depth seen on the ultrasound images was compared to the ES depth measured by the needle. We compared the number of puncture attempts with the standard method (control group) to the number of attempts under ultrasound guidance. RESULTS: Ultrasonography significantly improved operating conditions for epidural anaesthesia. The maximum VAS scores and patient acceptance were significantly better. CONCLUSIONS: With ultrasound measurement of the ES depth, the quality of epidural anaesthesia was enhanced. PMID- 11421839 TI - Posture-related distribution of hyperbaric bupivacaine in cerebro-spinal fluid is influenced by spinal needle characteristics. AB - BACKGROUND: No studies have evaluated the relationship between duration of time sitting and spinal needle type on the maximal spread of local anaesthetics. The few trials available have studied the influence of time spent sitting on the spread of anaesthesia without standardising spinal needle types, and have not found any effect. METHODS: In this randomised, blinded study, 60 patients scheduled for elective orthopaedic surgery of the lower limbs were divided into 4 groups. With the patient sitting erect, 15 mg hyperbaric bupivacaine were injected in a standard manner through a 24G Sprotte or a 27G Whitacre needle and patients were placed supine after 1 min (24G/1 group and 27G/1 group) or 4 min (24G/4 group and 27G/4 group). RESULTS: Time to achieve maximum block height after injection was similar in all groups. Block height levels were significantly lower at all time points for the 24G/4 group. Maximum block heights were Th4 in the 24G/1, 27G/1 and 27G/4 groups, and Th6 in the 24G/4 group (P<0.0001). CONCLUSION: In a standard spinal anaesthesia procedure, when different lengths of time spent sitting are compared, spinal needle characteristics influence the maximum spread of hyperbaric bupivacaine. However, within the limits of our study, a two-segment difference in block height is too small to consider using spinal needles as valuable tools to control block height during spinal anaesthesia in our daily practice. PMID- 11421840 TI - Effects of hypertonic 75 mg/ml (7.5%) saline on extracellular water volume when used for preloading before spinal anaesthesia. AB - BACKGROUND: Prevention of hypotension during spinal anaesthesia is commonly achieved using fluid preloading. This may result in a substantial amount of excess free water retained in the body after spinal anaesthesia. We aimed to evaluate the effects of 7.5% hypertonic saline on extracellular water volume and haemodynamics when used for fluid preloading before spinal anaesthesia. METHODS: This randomised double-blind study evaluated the effects of 75 mg/ml (7.5%) hypertonic saline (HS) on extracellular water volume and haematocrit in patients undergoing arthroscopy or other lower limb surgery under spinal anaesthesia. Amounts of 1.6 ml/kg of HS (20 patients) or 13 ml/kg of 9 mg/ml normal saline (20 patients) were administered for preloading before spinal anaesthesia with a 10 mg dose of 0.5% hyperbaric bupivacaine. Etilefrine was administered in order to maintain mean arterial pressure (MAP) at >or=80% of its baseline value. Whole body impedance cardiography-derived cardiac index (CI) and extracellular water (ECW) were measured. RESULTS: There were no significant differences in demographic data or in the number of blocked segments. ECW remained similar in both groups despite the much smaller amount of infused free water in the HS group. There were no significant differences between the groups in CI values during the study. The amount of etilefrine administered was similar in the treatment groups. Dilution of haematocrit was also similar in both groups. CONCLUSION: Hypertonic 75 mg/ml (7.5%) saline is an alternative for preloading before spinal anaesthesia in situations where excess free water administration is not desired. It is effective in small doses of 1.6 ml/kg, which increase the extracellular water, plasma volume and cardiac output, and thus maintain haemodynamic stability during spinal anaesthesia. PMID- 11421841 TI - Postoperative analgesia by combined continuous infusion and patient-controlled epidural analgesia (PCEA) following hip replacement: ropivacaine versus bupivacaine. AB - BACKGROUND: Ropivacaine is a new local anaesthetic, which compared to bupivacaine is less toxic and shows greater sensory and motor block dissociation. We hypothesised that treatment of postoperative pain with a combined regimen of continuous epidural infusion and Patient-Controlled Epidural Analgesia (PCEA) using ropivacaine could have given better results compared with those we had obtained using bupivacaine. METHODS: Patients undergoing total hip replacement were randomly assigned to two groups. They received epidural analgesia for postoperative pain treatment using ropivacaine, 2 mg x ml(-1) or bupivacaine 2 mg x ml(-1). Both drugs were administered as a constant infusion of 6 ml x h(-1) supplemented by PCEA bolus doses of 2 ml. Patients in both groups received morphine intravenously on demand from a patient-controlled analgesia (PCA) device. An independent observer recorded pain scores, intensity of motor block and morphine consumption at regular intervals during the first 24 h after surgery. RESULTS: Fifty-one patients were evaluated. Ropivacaine and bupivacaine, in similar amounts, provided similar results assessed as adequate to very good postoperative analgesia, whereas motor block was significantly more intense in patients treated with bupivacaine. CONCLUSIONS: Despite similar analgesic effects, epidural infusion of ropivacaine combined with PCEA provides higher patient satisfaction than equal doses of bupivacaine due to lack of motor block. PMID- 11421842 TI - Caudal bupivacaine-tramadol combination for postoperative analgesia in pediatric herniorrhaphy. AB - BACKGROUND: Administration of bupivacaine caudally has been used for postoperative analgesia after urogenital, rectal and lower abdominal surgery in children. Caudal opioids may offer analgesic advantages over bupivacaine alone but have been associated with side effects such as respiratory depression. Tramadol is an analgesic assumed to lack a respiratory depressant effect and has been shown to provide effective, long-lasting analgesia after epidural administration in adults and children. The aim of this study was to determine whether the addition of tramadol to bupivacaine caudally prolongs the duration of analgesia compared with bupivacaine alone, with respect to side effects, and whether caudal tramadol alone provides satisfactory analgesia. METHODS: Sixty boys, aged 12-84 months, undergoing unilateral herniorrhaphy, were allocated randomly to three groups. Children in group B received 0.25% plain bupivacaine 1 ml kg(-1), group BT received an identical local anesthetic dose mixed with tramadol 1.5 mg kg(-1) and group T received caudal tramadol 1.5 mg kg(-1) in 0.9% sodium chloride in the same total volume (1 ml kg(-1)). Pain and demeanour assessments were made 1, 2, 3, 4, 6, 12 and 24 h after recovery from anesthesia with reference to a three-point scale. RESULTS: Analgesia time (time between caudal injection and first administration of analgesic) in group BT (13.5+/-2.2 h) was significantly longer than in the other two groups (P<0.05). In group T, more patients required additional analgesia after surgery than in the other two groups (P<0.05). Pain scores in the three groups were similar up to 4 h after operation but the mean score in group T was higher than groups B and BT 4 and 6 h after operation (P<0.05). Significantly more patients who had received caudal bupivacaine alone or with tramadol had lower pain and demeanour scores during the first 24 h after operation compared with those in the tramadol group. CONCLUSION: Caudal administration of bupivacaine with the addition of tramadol resulted in superior analgesia with a longer period without demand for additional analgesics compared with caudal bupivacaine and tramadol alone without an increase of side effects. PMID- 11421843 TI - Effects of changes in frequency and inspiratory time on arterial oxygenation and CO2 elimination during high-frequency jet ventilation in a child with laryngotracheal papillomata. AB - An 11-yr-old female without any pulmonary disorders underwent laser resections of laryngotracheal tumors using high-frequency jet ventilation (HFJV) 6 times over a period of 17 months at our institute. In this series of surgeries, we studied the effects on PaO2 and PaCO2 during HFJV of changes of either frequency or inspiratory time. Increasing the frequency from 100 to 400/min decreased the PaO2 and increased the PaCO2. Decreasing the inspiratory time from 30% to 20% increased the PaCO2, although it did not affect the PaO2. All the procedures were uneventfully carried out without critically impairing gas exchange during HFJV. In this patient with normal lung function, CO2 elimination during HFJV appeared to be facilitated by either decreasing the frequency or increasing the inspiratory time. Arterial oxygen tension during HFJV was higher at lower frequencies. PMID- 11421844 TI - Hot chili, a cook and EMLA. PMID- 11421845 TI - Colorectal symptoms in patients with neurological diseases. AB - Several neurological diseases cause constipation or faecal incontinence restricting social activities and influencing quality of life. As several new treatment modalities have become available within the last few years, doctors treating patients with neurological diseases should be aware of the symptoms and have a basic knowledge of relevant treatment options. Constipation and faecal incontinence are common symptoms in patients with traumatic spinal cord injuries, spina bifida, multiple sclerosis, diabetic polyneuropathy, Parkinson's disease, stroke, and cerebral palsy. New treatment modalities are: prokinetic agents, enemas administered through the enema continence catheter or through an appendicostomy, and biofeed-back. Sacral nerve stimulation is still mostly at an experimental level and colostomy should be restricted to the few patients with most severe problems resistant to other therapy. PMID- 11421846 TI - Prognostic factors in the surgical treatment of medically intractable epilepsy associated with mesial temporal sclerosis. AB - OBJECTIVES: To assess the prognostic factors determining seizure remission after temporal lobectomy for intractable epilepsy associated with mesial temporal sclerosis (MTS) at pathology. METHODS: The clinical and investigative features of 116 consecutive patients who had temporal lobe surgery for drug-resistant epilepsy and MTS at pathology were assessed using actuarial statistics and logistic regression analysis. RESULTS: At a median follow-up of 63 months the probability of achieving at least a 1-year period of continuous seizure freedom was 67%. Factors contributing to a favourable outcome were interictal EEG localization to the operated lobe and the absence of secondarily generalized seizures. These were also selected in the multivariate analysis, although at lower statistical significance (P=0.08 and 0.09, respectively). Perinatal complications were associated with a significantly worse outcome but overall, complicated febrile convulsions and congruent neuropsychological deficits were not significantly predictive variables. CONCLUSIONS: The present findings may aid in the non-invasive presurgical assessment of patients with intractable TLE and clinical and neuroimaging evidence of MTS. PMID- 11421847 TI - Pre--and postoperative general neurocognitive status and memory in 70 epilepsy surgery patients. AB - PURPOSE: To describe cognitive function preoperatively and cognitive change 2 years postoperatively in 70 consecutive adults in the Goteborg epilepsy surgery series. METHODS: Analysis of general neurocognitive status and memory for the total group and for three subgroups: 26 patients with seizure onset in dominant temporal lobe, 28 patients with onset in non-dominant temporal lobe and 12 with frontal lobe onset. Preoperative performance was compared with a control group (n=30). RESULTS: Preoperatively, patients were more impaired in general neurocognitive status than controls. Patients in the temporal lobe subgroups performed worse than controls in two memory variables. No postoperative negative change was found at group level. There was a small increase in Full Scale IQ and Performance IQ for the total patient group, and in Performance IQ for the frontal lobe subgroup. At individual level, many patients suffered losses or obtained gains in a number of memory variables without any consistent intra-pattern. CONCLUSIONS: Cognitive stability was the main postoperative result. The importance of considering inter- and intra-individual variability of negative as well as positive change is emphasized. PMID- 11421848 TI - PET with 11C-deuterium-deprenyl and 18F-FDG in focal epilepsy. AB - OBJECTIVES: This study compares positron emission tomography (PET) using 11C deuterium-deprenyl (DED) with PET using 18F-fluorodeoxyglucose(18F-FDG) for examining epileptogenic regions in patients with focal epilepsy. MATERIAL AND METHODS: Twenty-three patients undergoing evaluation for epilepsy surgery were subjected to PET with DED. Fourteen patients had mesial temporal lobe epilepsy (TLE) and 9 patients had seizures of neocortical origin. In addition, 6 healthy control subjects were examined. Pixel-by-pixel analysis was used to generate graphical images of tracer distribution volume (intercept) and the accumulation rate (slope). Asymmetries with respect to relative intercept and slope were compared in patients with temporal lobe epilepsy (TLE), in patients with extra temporal lobe epilepsy (exTLE), and in the control subjects. The results were compared with 18F-FDG-PET. RESULTS: Among the patients with TLE, significant differences between the epileptogenic and the contralateral lobe were found with DED intercept and FDG-uptake. No significant differences were found with DED slope. The exTLE and the control groups showed no significant differences between sides or lobes. CONCLUSIONS: This study indicates that PET with 11C-deuterium deprenyl is a useful method for identifying TLE and is equivalent to PET with 18F FDG in this sense. The method has little localizing value in seizures originating from neocortical structures. PMID- 11421849 TI - Behaviour in frontotemporal dementia, Alzheimer's disease and vascular dementia. AB - OBJECTIVES: The study aimed to increase understanding of behavioural changes in frontotemporal dementia (FTD) and identify features that best differentiate FTD from Alzheimer's disease (AD) and cerebrovascular dementia (CvD). METHODS: A semi structured questionnaire was administered to carers of 30 FTD, 75 AD and 34 CvD patients. RESULTS: Behavioural changes that strongly discriminated FTD from AD and to a lesser extent CvD were loss of emotions and insight, selfishness, disinhibition, personal neglect, gluttony and sweet food preference, wandering, motor and verbal stereotypies, loss of pain, echolalia and mutism. Irritability, hyposexuality and hypersomnia did not discriminate. Emotional, eating and stereotyped behaviours correctly classified 95% of patients using regression analysis. CONCLUSIONS: Behavioural characteristics accurately differentiate FTD from AD and CvD. The findings highlight the particular importance of affective change in FTD, and underline the role of the frontotemporal lobes in emotion. PMID- 11421850 TI - Astheno-emotional disorder after aneurysmal SAH: reliability, symptomatology and relation to outcome. AB - OBJECTIVES: Neuropsychiatric symptoms commonly found after aneurysmal SAH are covered in the astheno-emotional disorder (AED) of Lindqvist & Malmgrens diagnostic system for organic psychiatry. This study aims to describe the reliability and symptomatology of AED and its relationship with social outcome. MATERIAL AND METHODS: Patients referred due to aneurysmal SAH (n=78) were assessed after 1-6 months for AED inter-rater reliability (n=36) and after 12 months for AED severity grade, symptomatology and Glasgow Outcome Scale (GOS) (n=63). RESULTS: There was no systematic error in diagnosing and grading AED, and the agreement was 85% and 67% respectively (kappa=0.65 and 0.52). Fatigability, concentration difficulties and memory difficulties were the most frequent symptoms of AED. The relationship between AED and "moderate disability" of GOS was highly significant (P<0.00006). CONCLUSIONS: AED affects social outcome, and can be diagnosed with high reliability even without neuropsychological testing. Use of the AED diagnosis for evaluation of organic mental symptoms is encouraged. PMID- 11421851 TI - Peripheral neuropathy in systemic lupus erythematosus--a longitudinal study. AB - OBJECTIVE: Peripheral neuropathy (PN) is reported to occur in 5-27% of patients with systemic lupus erythematosus (SLE) mostly as a length-dependent sensorimotor axonopathy. Studies over time have not been performed. Design - Longitudinal study. SUBJECTS AND METHODS: Thirty-three Caucasian SLE patients consented to participate in the study and were subjected to clinical examination, laboratory tests, and nerve conduction velocity (NCV) studies. At the follow-up 7 years later, 7 patients (21%) were dead, 4 refused to participate, and 2 did not want to perform NCV studies. Twenty patients were thus available for longitudinal study. RESULTS: When all SLE patients were considered on a group basis at follow up, 8 (33%) out of 24 NCV parameters showed significant deterioration despite correction for time, while 16 (67%) were unchanged. Analysis of change from baseline showed that, except for F-responses, several NCV changes were highly dependent (negative regression coefficients) on baseline levels at start of study. No demographic, laboratory, or disease associated quantitative factor was associated with these changes in NCV parameters over time. Nor was a consistent effect on NCV parameters from any qualitative demographic or disease associated factor confirmed by Repeated Measures ANOVA analyses. CONCLUSIONS: A modest progressive neuropathic process exists in patients with SLE. Important is also the finding that, over time, the abnormalities of NCV parameters fluctuate in the individual patients, and the impairments are not necessarily irreversible. This study also shows no association to medication, demographic-, or other disease associated factors. PMID- 11421852 TI - Maximal respiratory pressures in myasthenia gravis. Relation to single fiber electromyography. AB - OBJECTIVES: The respiratory muscle weakness was evaluated in mild forms of myasthenia gravis (MG) and was compared with single fiber electromyography results (SFEMG) in the extensor digitorum communis muscle. MATERIAL AND METHODS: We assessed 61 treated MG patients without clinical respiratory involvement (13 ocular forms, 28 form IIa and 20 form IIb patients) by maximal respiratory pressure (MRP) to ascertain whether they were consistent with SFEMG results. RESULTS: Maximal expiratory pressure (MEP) and maximal inspiratory pressure (MIP) were reduced below the lower limit for healthy subjects in 85% and 77% of ocular patients; in 93% and 89% of form IIa, respectively; and in 95% of form IIb. The highest percentage of MEP and MIP reduction showed a positive correlation with increased jitter (r=0.45, P<0.001; r=0.27; P<0.05 respectively) and impulse blocking (r=0.35, P<0.01; r=0.28, P<0.05 respectively). CONCLUSION: We consider MRP measure useful for assessing respiratory muscle weakness in MG patients with subclinical respiratory involvement. PMID- 11421853 TI - Complications after LP related to needle type: pencil-point versus Quincke. AB - OBJECTIVES: We studied the incidence of complications after diagnostic lumbar puncture (LP) related to needle type. MATERIAL AND METHODS: A 5 months' observational study of routine diagnostic LP in 83 patients was conducted. RESULTS: Significantly more headache was observed after LP using thicker cutting needles (20G Quincke) compared with thinner cutting or non-cutting needles (22G Quincke or pencil-point). No significant difference in complications after LP was found between the 22G Quincke and pencil-point needles. CONCLUSION: The size of the needle and not the needle shape seems to be the main determinant for post dural puncture headache (PDPH). PMID- 11421854 TI - Regional cardiac expression and concentration of natriuretic peptides in patients with severe chronic heart failure. AB - The purpose of this study was to examine the regional cardiac mRNA expression and concentration of brain natriuretic peptide (BNP) and atrial natriuretic peptide (ANP) in relation to the circulating peptide concentrations in patients with chronic heart failure (CHF). The myocardial mRNA levels and peptide concentrations of BNP and ANP were analysed in seven different regions of the heart from patients undergoing cardiac transplantation. Autopsy samples from individuals without known cardiovascular disease were used as controls. The plasma levels of natriuretic peptides and their N-terminal propeptides, Nt-proBNP and Nt-proANP, were measured in the CHF patients and healthy volunteers. In the autopsy specimens, the atrial regions appeared to contain the highest peptide levels for BNP as well as ANP, the atrioventricular ratio being 12-262 and 72-637 fold, respectively. In the CHF patients there was a relative shift towards the ventricle for BNP, reducing the atrioventricular ratio to 6-16-fold. The circulating concentrations of BNP/Nt-proBNP in the CHF patients correlated closely to the BNP mRNA expression in most myocardial regions including the left ventricle (r = 0.72, P < 0.001). For circulating concentrations of ANP/Nt-proANP, such correlation were limited to the left atrium free wall (r = .66, P < 0.002). Thus, of the two natriuretic peptides, BNP/Nt-proBNP may be a better reflector of left ventricular overload. PMID- 11421855 TI - Mechanisms of hypotensive effects of a posture change from seated to supine in humans. AB - The hypothesis tested was that the hydrostatic stimulation of carotid baroreceptors is pivotal to decrease mean arterial pressure at heart level during a posture change from seated to supine. In eight males, the cardiovascular responses to a 15-min posture change from seated to supine were compared with those of water immersion to the xiphoid process and to the neck, respectively. Left atrial diameter and cardiac output (rebreathing) increased similarly during the posture change and water immersion to the xiphoid process and further so during neck immersion. Mean arterial pressure decreased by 12 +/- 2 mmHg during the posture change, by 5 +/- 1 mmHg during xiphoid immersion, and was unchanged during neck immersion. Arterial pulse pressure increased by 12 +/- 3 mmHg during the posture change (P < 0.05) and less during xiphoid and neck immersion by 7 +/- 3 mmHg (P < 0.05). Total peripheral vascular resistance decreased similarly during the posture change and neck immersion and slightly less during xiphoid immersion (P < 0.05). In conclusion, the hydrostatic stimulation of carotid baroreceptors combined with some additional increase in arterial pulse pressure, which also stimulates aortic baroreceptors, accounts for more than half of the hypotensive response at heart level to a posture change from seated to supine. PMID- 11421856 TI - Nitric oxide modulates acetylcholine-induced vasodilatation in the hepatic arterial vasculature of the dual-perfused rat liver. AB - The role of nitric oxide in the modulation of hepatic arterial vascular reactivity was investigated in an isolated dual-perfused rat liver preparation. Twelve male Wistar rats (200-250 g) were anaesthetized with sodium pentobarbitone (60 mg kg-1 i.p.). The livers were then excised and perfused in vitro through hepatic arterial and portal venous cannulae at constant flow rates. Concentration dependent dose-response curves to acetylcholine (10(-8)-10(-5) M), sodium nitroprusside (10-6(-5) x 10(-4) M), and adenosine triphosphate (ATP) (10(-8)-10( 5) M) in the hepatic artery were constructed after the tone was raised by addition of methoxamine (3 micorM L(-1)). Acetylcholine-induced vasodilatation in the hepatic artery was significantly attenuated with inhibition of nitric oxide synthase by using NG-nitro-L-arginine methyl ester (30 microM), Emax = 51.7 +/- 2.8 vs. 32.5 +/- 3.1 mmHg, before vs. after NG-nitro-L-arginine methyl ester, respectively. ATP-induced hepatic arterial vasoconstriction which was significantly enhanced with L-NAME, Emax = 94.0 +/- 9.3 vs. 127.0 +/- 8.0 mmHg, before vs. after NG-nitro-L-arginine methyl ester, respectively. Sodium nitroprusside-induced hepatic arterial vasodilatation remained unchanged with NG nitro-L-arginine methyl ester, Emax = 57.0 +/- 3.4 vs. 57.0 +/- 4.1, before vs. after NG-nitro-L-arginine methyl ester, respectively. The data from the present study suggest that acetylcholine-induced vasodilatation in the intrahepatic arterial vasculature of the rat liver is at least, in part, mediated by the release of nitric oxide. In addition, ATP-induced hepatic arterial vasoconstriction is also modulated by the release of nitric oxide (*P < 0.05, Student's paired t-test). PMID- 11421857 TI - Effects of lactoferrin on rat dermal interstitial fluid pressure (Pif) and in vitro endothelial barrier function. AB - We recently demonstrated that intravenous (i.v.) injection of the iron-binding protein lactoferrin (Lf) followed by antilactoferrin (aLf) antibodies or iron saturated Lf alone increased albumin extravasation in vivo in several tissues including skin. Increased driving pressure for blood-tissue exchange or direct effects of Lf on the endothelial barrier are possible mechanisms. We therefore, firstly, measured interstitial fluid pressure (Pif) in dermis of rats given 1 mg Lf i.v. followed 30 min later by aLf or saline and circulatory arrest 1 or 5 min thereafter and compared with controls. Secondly, transmonolayer passage of Evans blue labelled albumin (EB-albumin) was evaluated in porcine pulmonary artery endothelial cells exposed to iron-free or iron-saturated Lf (both 100 microg mL 1) in the absence and presence of 0.5 mM hydrogen peroxide. Pif increased significantly at 11-30 min following Lf to +2.1 +/- 0.3 and +1.7 +/- 0.2 mmHg at 11-20 and 21-30 min, respectively, compared with +0.1 +/- 0.2 mmHg before Lf (P < 0.05, n=25). Endothelial transmonolayer passage of EB-albumin during 3 h was not affected by iron-free or iron-saturated Lf neither in the absence nor presence of hydrogen peroxide that increased passage 3.5 times compared with controls. In conclusion, Lf-induced increase in albumin extravasation in rat skin is not explained by changes in Pif (because Lf raised Pif significantly) or direct effects of Lf on the endothelial barrier. PMID- 11421858 TI - Insulin insensitivity and delayed transcapillary delivery of insulin in oophorectomized rats treated with testosterone. AB - The importance of transcapillary insulin delivery as a regulated step was explored in an insulin resistant rat model. Oophorectomized female rats were exposed to testosterone (OVX + T) for 8 weeks and examined with insulin clamps, muscle microdialysis, and analyses of insulin distribution kinetics. The results were compared with those obtained in sham-operated control rats. After OVX + T, onset of glucose uptake in skeletal muscle was significantly (P < 0.001-0.05) delayed compared with controls as measured by the glucose infusion rate (GIR) during a euglycaemic, hyperinsulinaemic clamp (5 mU kg-1 min-1). The increase in interstitial insulin concentrations was also significantly (P < 0.05) delayed (15 20% lower) in OVX + T treated rats compared with control rats, but to such a small magnitude that this alone could not explain the late onset of the insulin effect. Skeletal muscle capillary density, examined histochemically, was diminished (P < 0.01-0.001) by 20-25% after treatment with OVX + T compared with control animals, as was the peripheral blood flow (P < 0.05) by 40-45%, measured with the microsphere technique. Insulin binding was reduced in proportion to the reduced (P < 0.01) vascular surface area by OVX + T treatment. Transcapillary transport rate of insulin, measured by comparisons of the kinetics of inulin and insulin spaces in muscle with time, tended (ns) to be lower after OVX + T compared with control rats (30-40%) as a reflection of the lower capillary surface area. The data suggest that the delayed onset of insulin action after OVX + T results from combined defects in the muscle cell at a postreceptor level and, to a lesser extent, from retarded transcapillary delivery of insulin. PMID- 11421859 TI - Effect of force development on contraction induced glucose transport in fast twitch rat muscle. AB - A previous study has shown that in fast twitch frog sartorius muscle contraction stimulated glucose transport depends only on stimulation frequency and not on workload. In contrast, we have recently shown that in rat slow twitch muscle stimulated to contract at constant frequency, glucose transport varies directly with force development and, in turn, metabolism. The present study was carried out to clarify whether the discrepancy between the earlier studies reflected differences in physiological behaviour between fast and slow twitch muscle. We investigated the effect of force development on glucose transport in incubated fast twitch rat flexor digitorum brevis (rich in type 2a fibres) and epitrochlearis (rich in type 2b fibres) muscle. Muscles were electrically stimulated to perform repeated tetanic contractions at 1 Hz for 10 min. Resting length was adjusted to achieve either no force or maximum force. Glucose transport (2-deoxyglucose uptake) increased when force was produced compared with when it was not (P < 0.05) in both flexor digitorum brevis (19 +/- 7 (basal), 163 +/- 14 (no force) and 242 +/- 17 (max force) nmol x g(-1) x 5 min(-1)) and epitrochlearis (60 +/- 4 (basal), 100 +/- 7 (no force) and 125 +/- 6 (max force) nmol x g(-1) x 5 min(-1)). In both muscles glucose transport increased in parallel with metabolic rate, as reflected by muscle lactate concentrations and 5' AMP-activated protein kinase activity, during contractions. In conclusion, as previously shown for rat soleus muscle, at a given stimulation frequency glucose transport varies directly with force development in rat flexor digitorum brevis and epitrochlearis muscle. Accordingly, force development enhances glucose transport in all mammalian muscle fibre types. The influence of force development probably reflects effects of enhanced 5' AMP-activated protein kinase activity resulting from reduced intra-cellular energy status and pH. PMID- 11421860 TI - Cell volume-induced changes in K+ transport across the rat colon. AB - The effect of cell swelling and cell shrinkage on K+ transport across the rat colonic epithelium was studied by measuring unidirectional fluxes, uptake and efflux of 86Rb+, a marker for K+. Exposure to a hypotonic medium stimulated the secretory, serosa-to-mucosa flux of K+, whereas exposure to a hypertonic medium inhibited the absorptive, mucosa-to-serosa flux of K+ in the distal, but not in the proximal colon. Neither manoeuvre had any effect on the uptake of K+ across the apical or the basolateral membrane. Cell swelling induced a sustained increase in the apical and basolateral K+ efflux from both colonic segments, whereas cell shrinkage reduced the efflux. Ba2+ (10(-2) mol l(-1)) inhibited the swelling-induced stimulation of the apical, quinine (10(-3) mol l(-1)) that of the basolateral K+ efflux in the distal colon. Incubation of the tissue in Ca2+ free buffer or La3+, which blocks Ca2+-influx into the epithelium, strongly reduced the basal K+ efflux across the basolateral membrane. The same was observed with brefeldin A, a blocker of the transport of newly synthesized proteins out of the endoplasmatic reticulum. Swelling-induced K+ efflux, however, was not reduced. In the presence of colchicine, an inhibitor of the polymerization of microtubules, swelling evoked only a transient increase in mucosal efflux, which, especially in the proximal colon, fell after 6 min to the level of the isotonic control period. These results demonstrate that the cell volume is involved in the regulation of transepithelial K+ transport across the rat colonic epithelium and suggest a role of the cytoskeleton in the control of a part of the volume-sensitive K+ channels. PMID- 11421861 TI - Caffeine-induced depolarization in amphibian skeletal muscle fibres: role of Na+/Ca2+ exchange and K+ release. AB - Caffeine (4 mM) produces a depolarization of about 10 mV in frog muscle fibres (Leptodactylus ocellatus). The aim of this work was to study the mechanisms of this effect. An approximately threefold rise in membrane resistance [Cl--free (SO(4)2-) medium] substantially increased, and both Na+-free medium and Ni2+ (5 mM) reduced, the caffeine-induced depolarization. In voltage-clamped (-60 mV) short fibres from lumbricalis muscle of the toad (Buffo arenarum), caffeine generated an inward current of 4.13 +/- 0.48 microA cm(-2). This caffeine-induced current was reduced by 60% in Na+-free medium, 44% in the presence of 5 mM amiloride and 48% by 5 mM Ni2+, suggesting that the activation of the Na+-Ca2+ exchanger in its forward mode may play a role in the observed electrical effects of the drug. Caffeine also produced a marked release of K+. Net K+ efflux increased from 3.5 +/- 0.2 (control) to 22.1 +/- 2.3 pmol s(-1) cm(-2) (caffeine). It is shown that in the presence of the drug, [K+] in the lumen of the T tubules may well increase to levels which could produce, in part, both the observed depolarization and the caffeine-induced current under voltage clamp conditions. The caffeine-induced K+ efflux was not reduced by 5 mM Ni2+. At a holding potential of 30 mV the caffeine-induced current was reversed (outward) and roughly halved by 5 mM Ni2+. The Ni2+-sensitive fraction of the caffeine induced current, assumed to represent the Na+-Ca2+ exchanger current, had an estimated reversal potential close to 12 mV ([Na+]o = 115 mM; [Ca2+]o = 1 mM). In conclusion, the depolarizing effect of caffeine described here would be produced by two mechanisms: (a) an inward current generated by the activation of the Na+ Ca2+ exchanger in its forward mode, and (b) the rise of the external [K+] in restricted spaces like the T tubules. PMID- 11421862 TI - Infusion of vasopressin is associated with increased milk flow in the goat. PMID- 11421863 TI - The pharmacology of the internal anal sphincter and new treatments of ano-rectal disorders. AB - Surgical options for faecal incontinence in the presence of intact sphincters are limited. Furthermore, in patients with fissures, lateral sphincterotomy reduces anal sphincter hypertonia but there has been concern about complications. A greater understanding of the basic pharmacology of the internal anal sphincter has led to the development of novel treatments for both these disorders. A Medline review was undertaken for internal anal sphincter pharmacology, anal fissures and faecal incontinence. This review is based on these articles and those found by further cross-referencing. Nitric oxide released from non adrenergic non-cholinergic nerves is the main inhibitory agent in the internal anal sphincter. Relaxations are also mediated through beta-adrenoceptors and muscarinic receptors. Stimulation of alpha-receptors results in contraction. Calcium and its entry through L-type calcium channels is important for the maintenance of tone. Nitric oxide donors produce reductions in resting anal tone and heal fissures but are associated with side-effects. Muscarinic agents and calcium channel antagonists show promise as low side-effect alternatives. Botulinum toxin appears more efficacious than other agents in healing fissures. To date, alpha-receptor agonists have been disappointing at improving incontinence. Further understanding of the pharmacology of the internal anal sphincter may permit the development of new agents to selectively target the tissue with greater efficacy and fewer side-effects. PMID- 11421864 TI - Photodynamic therapy and the alimentary tract. AB - Photodynamic therapy offers the possibility of relatively selective tumour necrosis and normal tissue healing. It has many potential applications but as yet no clear role. Articles, editorials and case reports published primarily in English and listed in Medline/ISI up to April 2000 or identified by a manual search have been reviewed in an attempt to provide a comprehensive overview of the use of photodynamic therapy in the alimentary tract. It is concluded that photodynamic therapy can be an effective treatment for superficial pre-malignant mucosal lesions and early cancers, especially in diffuse disease. Suitable patients include those wishing to avoid surgery, high risk subjects or those in whom other forms of treatment have failed. Superiority over other methods of ablation has not so far been demonstrated. Cheaper and more effective photosensitizers and improved techniques of light delivery are likely to increase the application of photodynamic therapy. PMID- 11421865 TI - Proton pump inhibitors versus H2-antagonists: a meta-analysis of their efficacy in treating bleeding peptic ulcer. AB - PURPOSE: To evaluate whether proton pump inhibitors are more effective than H2 antagonists (H2-A) for the treatment of bleeding peptic ulcer. DATA SOURCES: PubMed database until January 2000. STUDY SELECTION: Comparative randomized trials of proton pump inhibitors (omeprazole, lansoprazole, or pantoprazole) vs. H2-A (cimetidine, ranitidine or famotidine). DATA EXTRACTION: Meta-analysis combining the odds ratios (OR) of the individual studies in a global OR (Peto method). OUTCOMES EVALUATED: Persistent or recurrent bleeding, need for surgery, or mortality. DATA SYNTHESIS: Eleven studies fulfilled the inclusion criteria and contained data for at least one of the planned comparisons. Persistent or recurrent bleeding was reported in 6.7% (95% CI: 4.9-8.6%) of the patients treated with proton pump inhibitors, and in 13.4% (95% CI: 10.8-16%) of those treated with H2-A (OR 0.4; 95% CI: 0.27-0.59) (chi2-homogeneity test, 18; P=0.09). Surgery was needed in 5.2% (95% CI: 3.4-6.9%) of the patients treated with proton pump inhibitors, and in 6.9% (95% CI: 4.9-8.9%) of the patients treated with H2-A (OR 0.7; 95% CI: 0.43-1.13). Respective percentages for mortality were 1.6% (95% CI: 0.9-2.9%) and 2.2% (95% CI: 1.3-3.7%) (OR 0.69; 95% CI: 0.31-1.57). SUB-ANALYSIS: Five studies evaluated the effect of both therapies given in bolus injections on persistent or recurrent bleeding rate, which was 6% (95% CI: 3.6-8.3%) and 8.1% (95% CI: 5.3-10.9%), respectively (OR, 0.57; 95% CI: 0.31-1.05). Persistent or recurrent bleeding in high risk patients (Forrest Ia, Ib and IIa) occurred in 13.2% (95% CI: 7.9-8%) of the patients treated with proton pump inhibitors and in 34.5% (27-42%) of those treated with H2-A (OR 0.28; 95% CI: 0.16-0.48). In patients not having endoscopic therapy, persistent or recurrent bleeding was reported, respectively, in 4.3% (95% CI: 2.7-6.7%) and in 12% (95% CI: 8.7-15%) (OR 0.24; 95% CI: 0.13-0.43). Less marked differences were observed in patients having adjunct endoscopic therapy: 10.3% (95% CI: 6.7-13.8%) and 15.2% (11.1-19.3%) (OR 0.59; 95% CI: 0.36-0.97). Moreover, the significance disappeared in this group when a single outlier study was excluded. CONCLUSIONS: Proton pump inhibitors are more effective than H2-A in preventing persistent or recurrent bleeding from peptic ulcer, although this advantage seems to be more evident in patients not having adjunct sclerosis therapy. This beneficial effect seems to be similar or even more marked in patients with Forrest Ia, Ib or IIa ulcers. However, proton pump inhibitors are not more effective than H2-A for reducing surgery or mortality rates. Nevertheless, the data are too scarce and heterogeneous to draw definitive conclusions, and further comparative trials are clearly warranted. PMID- 11421866 TI - The new proton pump inhibitor esomeprazole is effective as a maintenance therapy in GERD patients with healed erosive oesophagitis: a 6-month, randomized, double blind, placebo-controlled study of efficacy and safety. AB - BACKGROUND: Esomeprazole, the S-isomer of omeprazole, is the first proton pump inhibitor to be developed as an optical isomer. In patients with erosive oesophagitis, esomeprazole has produced significantly greater healing rates and improved symptom resolution vs. omeprazole. AIM: This study assesses the efficacy of esomeprazole for preventing relapse in patients with healed oesophagitis. METHODS: In this 6-month US multicentre randomized double-blind placebo controlled trial, 375 Helicobacter pylori-negative patients with endoscopically healed oesophagitis received esomeprazole 40 mg, 20 mg, 10 mg, or placebo once daily. The primary efficacy end-point was maintenance of healing at 6 months. Secondary end-points assessed changes in symptoms, and long-term safety and tolerability. RESULTS: Significantly (P < 0.001) more patients remained healed with esomeprazole 40 mg (87.9%), 20 mg (78.7%), or 10 mg (54.2%), than with placebo (29.1%). Relapse, when it occurred, was later with esomeprazole. Sustained resolution of heartburn was observed in the 40 mg and 20 mg groups; there was a high correlation between absence of heartburn and maintenance of healing. Adverse effects were mild, infrequent and not significantly different between groups. CONCLUSIONS: Esomeprazole is effective and well-tolerated in the maintenance of healing of erosive oesophagitis. Esomeprazole 40 mg and 20 mg offer significant clinical benefit to patients. PMID- 11421867 TI - The pharmacokinetics of the novel promotile drug, tegaserod, are similar in healthy subjects-male and female, elderly and young. AB - BACKGROUND: Tegaserod (HTF 919) is a selective 5-HT4 receptor partial agonist in development for the treatment of irritable bowel syndrome. AIM: This study aimed to assess the effect of age and gender on the single-dose pharmacokinetics of tegaserod. METHODS: In a parallel-group, open-label study, a single dose of tegaserod (12 mg) was administered to four groups of healthy young male, young female, elderly male and elderly female subjects (n=10 per group). Blood samples were collected from 0 to 24 h postdose. Non-compartmental pharmacokinetics evaluation and statistical analysis (ANOVA and Wilcoxon signed ranks test for tmax) were performed. RESULTS: Tegaserod was well tolerated in all groups. There was no effect of age or gender on tmax and Cmax. Gender did not affect AUC0 infinity and AUC0-tz; there was a statistically significant effect of age on these parameters. AUC0-infinity and AUC0-tz in the elderly were greater than in the young (AUC0-infinity ratio 1.37, P < 0.001; AUC0-tz ratio 1.23, P=0.029). This increase in exposure is judged not to be clinically relevant because it is within the variability in the pharmacokinetics parameters of tegaserod and because the dose-response relationship of tegaserod is relatively shallow. CONCLUSIONS: No dose adjustment for age or gender is recommended in tegaserod therapy. PMID- 11421868 TI - Healthcare resource utilization in the management of oesophageal adenocarcinoma. AB - OBJECTIVE: To study the natural history of oesophageal adenocarcinoma in terms of utilization of the healthcare resources and associated medical costs. METHODS: All 29 patients treated at the University of New Mexico Health Center between 1 January 1992 and 1 December 1998 for an oesophageal adenocarcinoma were included in the study. For each individual patient, all medical resources utilized, facility costs, and physician fees were retrieved from the computerized databases of the collection departments. RESULTS: During the progress of the disease, the same type of diagnostic or therapeutic procedure became necessary a number of times, and patients underwent, on average, three (range one to eight) upper gastrointestinal endoscopies, five (range one to 11) computerized tomography scans, 35 (five to 143) X-ray examinations, and 28 (three to 46) radiation therapies or 39 (10 to 74) chemotherapies. The large variation in the numbers of resources utilized was also reflected by a corresponding variation in total healthcare costs. The mean cost per patient was $48 127, ranging between $13 454 and $139 721. Facility costs comprised 88% of all costs compared to physician fees which comprised 12%. The largest cost items, in declining order, were physician encounters ($16 916), radiation plus chemotherapy ($9909 plus $4891, respectively), and pharmacy prescriptions ($7565). CONCLUSIONS: The prolonged disease process and the many diagnostic and therapeutic procedures result in complications, side-effects, inconclusive tests, or failed therapies that all markedly increase the use of healthcare resources. Because such outcomes are the rule rather than the exception, the management of oesophageal adenocarcinoma is expensive. PMID- 11421869 TI - High-dose prolonged combination therapy in non-responders to interferon monotherapy for chronic hepatitis C. AB - BACKGROUND: Therapy of chronic hepatitis C non- responders to interferon monotherapy with standard doses of interferon plus ribavirin is usually ineffective. AIM: To evaluate the efficacy and tolerability of high-dose prolonged combination retreatment in non- responder patients. METHODS: Patients were retreated for 6 months with 6 MU alphaIFN on alternate days and 1000 or 1200 mg/day ribavirin. HCV-RNA negative patients continued therapy for an additional 6 months. RESULTS: Forty patients (29 males, mean age 49.7 years, 34 genotype 1b, 11 with F3 fibrosis) were treated. At 6 months, 20 (50%) patients were HCV-RNA negative but six of them discontinued therapy because of adverse events. A sustained response was achieved in 28% of patients (11/40). A sustained response was more frequent among patients with genotype non-1b than in those with genotype 1b (67 vs. 21%, P=0.005) and clearance of HCV-RNA in the first 3 months had a high predictive value for sustained response (100% of sustained responders vs. 24% of non-responders, P=0.0001). CONCLUSIONS: High-dose prolonged combination therapy in non-responders to IFN monotherapy leads to a higher rate of sustained response than the standard combination regimen. Tolerability may be a rate limiting factor. Maximal effectiveness can be predicted in patients with non-1b genotype and in those who clear HCV-RNA soon after starting retreatment. PMID- 11421870 TI - Characteristics of patients with irritable bowel syndrome recruited from three sources: implications for clinical trials. AB - BACKGROUND: Variation in the characteristics of irritable bowel syndrome patients recruited for clinical trials from different sources could affect their response and the generalizability of trial results. AIM: To describe and compare the characteristics of three different groups of irritable bowel syndrome patients recruited into a 'mock clinical trial.' METHODS: We enrolled 245 irritable bowel syndrome patients from three sources: (i) 121 from British primary practitioners; (ii) 72 from California newspaper advertisements; and (iii) 52 from a California gastroenterologist's practice. We obtained demographic, clinical, and Hospital Anxiety and Depression (HAD) Scale data. RESULTS: Most patients were young to middle-aged women; the majority reported symptoms for > 5 years in all three groups. Subject characteristics varied among the groups. Typically, primary care patients were anxious, smokers and daily alcohol drinkers who had sought care recently for irritable bowel syndrome and tried antispasmodic drugs. Their symptoms were intermediate in severity between those of the other two groups. Advertisement subjects were the oldest, most highly educated, most often depressed, and were least likely to have sought care recently for symptoms, which were almost uniformly only moderate in severity. Gastroenterologist patients tended to be anxious and had nearly all sought care recently for symptoms, which were the most severe and most likely to include all three pain-related Rome I criteria. CONCLUSION: Recruitment methodology affects important characteristics of an irritable bowel syndrome study group. PMID- 11421871 TI - Octreotide 24-h prophylaxis in patients at high risk for post-ERCP pancreatitis: results of a multicenter, randomized, controlled trial. AB - BACKGROUND: Pharmacological prophylaxis of post-ERCP pancreatitis is costly and not useful in most non-selected patients, in whom the incidence of pancreatitis is 5% or less. However, it could be useful and probably cost-effective, in patients at high risk for this complication, where the post-procedure pancreatitis rate is 10% and more. AIM: To assess the efficacy of octreotide in reducing the incidence and severity of post-ERCP pancreatitis and procedure related hospital stay, in subjects with known patient-related risk factors. METHODS: A total of 120 patients were randomly allocated to receive octreotide or not, in a multicentre, randomized, controlled trial. The drug was given subcutaneously, 200 microg t.d.s., starting 24 h before the ERCP procedure, in patients with either sphincter of Oddi dysfunction, or a history of relapsing pancreatitis or post-ERCP pancreatitis, or who were aged under 35 years, or who had a small common bile duct diameter (< 8 mm). RESULTS: A total of 114 patients (58 in the octreotide group and 56 in the control group) completed the trial. Post-procedure pancreatitis occurred in seven octreotide-treated patients (12.0%) and eight controls (14.3%). The two groups showed no significant differences in the incidence or severity of pancreatitis. Twenty-four hours after the procedure, severe hyperamylasemia (more than five times the upper normal limit) without pancreatic-like pain was recorded in three octreotide-treated patients (5.2%) and six controls (10.7%), the difference being not significant. CONCLUSION: Twenty four-hour prophylaxis with octreotide proved ineffective in preventing post-ERCP pancreatitis and in avoiding 24-h severe hyperamylasemia in high-risk patients. PMID- 11421872 TI - Impaired bioavailability of vitamin A in adults and children with persistent diarrhoea in Zambia. AB - BACKGROUND: We have previously demonstrated a strong relationship between low serum retinol concentration and mortality in Zambian AIDS patients with diarrhoea, but were unable to detect any benefit from oral micronutrient supplementation. AIM: To test the hypothesis that this is related to impaired availability of vitamin A, we analysed serum retinol concentration changes over 6 h following oral mega-dose therapy (60, 120 or 180 mg retinol). METHODS: Twenty four men without diarrhoea, 15 adults with persistent diarrhoea and 11 children (six girls, five boys) with persistent diarrhoea were studied. RESULTS: Men with persistent diarrhoea had lower baseline serum retinol concentrations (median 0.39 micromol/L, interquartile range 0.21-0.56) than controls (median 1.16 micromol/L, interquartile range 0.84-1.47; P=0.0003). After 60 mg retinol, the rise in serum retinol in HIV seropositive controls (median 0.63 micromol/L, interquartile range 0.35-0.77) did not differ significantly from that observed in HIV seronegative controls (median 0.35 micromol/L, interquartile range - 0.04-0.56; P=0.20). Increasing the dose to 120 mg or 180 mg retinol did not enhance the increase in serum retinol concentration. The increase in serum retinol was less in adults with persistent diarrhoea (median 0.25 micromol/L, interquartile range 0.04-0.35) and in children (median 0.11 micromol/L, interquartile range 0.04-0.46) than in men without diarrhoea (median 0.44 micromol/L, interquartile range 0.26-0.74; P=0.03). Adults and children with diarrhoea had greater losses of retinol in urine over a 24-h period than controls, but less than 1% of the ingested dose was excreted. CONCLUSIONS: These results suggest that persistent diarrhoea in this population is associated with reduced bioavailability of retinol. Further work is required to determine the metabolic fate of therapeutic doses of retinol and to determine appropriate replacement strategies for HIV infected individuals. PMID- 11421873 TI - Reliability and validity of the digestive health status instrument in samples of community, primary care, and gastroenterology patients. AB - AIM: We evaluated a previously reported digestive health status instrument in community, primary care, and gastroenterology practice populations. Multiple types of reliability, validity and responsiveness were assessed to determine the performance of the questionnaire. METHODS: Study populations included community, primary care and gastroenterology subjects. Psychometric analyses included internal consistency and test-retest reliability, criterion and construct validity, and responsiveness. RESULTS: Acceptable internal consistency was seen on all scales in all three populations. Test-retest reliability was excellent in a speciality population with reflux disease. Criterion validity was demonstrated by strong correlation of reflux scale scores and results on 24-h pH monitoring. Scale scores varied predictably in those receiving gastrointestinal tract imaging and according to diagnosis, indicating construct validity. The reflux scale and pain index were sensitive to change with treatment for reflux disease. Multi trait scaling analyses from the community sample revealed a structure equivalent to that reported from a primary care sample. CONCLUSIONS: The reliability and validity of the digestive health status instrument on multiple measures in multiple settings have been demonstrated. The instrument was responsive to change with treatment for reflux disease. The demonstrated robustness attests to the suitability for future studies and clinical application. PMID- 11421874 TI - Expression of hepatocyte growth factor, transforming growth factor alpha, apoptosis related proteins Bax and Bcl-2, and gastrin in human gastric cancer. AB - BACKGROUND: Gastric cancer is one of the most frequent neoplasms and a leading cause of the death world-wide. In recent years, epidemiological and animal studies demonstrated a link between gastric cancer and chronic infection with H. pylori. The exact mechanism responsible for the development of gastric cancer in H. pylori-infected patients still remains unclear. There is evidence that the up regulation of certain growth factors could play an important role in the promotion of the gastric carcinogenesis. AIMS: The present study was designed to determine the gene expression of major known growth factors such as transforming growth factor alpha (TGFalpha), hepatocyte growth factor (HGF) and gastrin in the gastric cancer tissue, the surrounding mucosa and, for comparison, in the normal gastric mucosa. Furthermore, the luminal and plasma levels of gastrin in patients with gastric cancer were determined. In addition, the gene and protein expressions of apoptosis-related proteins such as Bax and Bcl-2 were investigated by reverse transcription-polymerase chain reaction and Western blot. Twenty-five gastric cancer patients and 40 age- and gender-matched control subjects hospitalized with non-ulcer dyspepsia were included into this study. RESULTS: An overall H. pylori-seropositivity among gastric cancer patients was about 72% and was significantly higher than in the controls (56%). The prevalence of CagA positive strains was also significantly higher among gastric cancer patients than in controls (56% vs. 32%). The gene expression of HGF and TGFalpha was detected more frequently in gastric cancer tissue samples than in normal gastric mucosa (52% vs. 12% for HGF and 48% vs. 24% for TGFalpha). The extent of protein expression in Western blotting analysis for HGF and TGFalpha correlated with the mRNA expression of these factors. Gene expression of gastrin was detected in the antrum of all tested patients and in the majority (84%) of gastric cancer patients. The median plasma and luminal concentrations of gastrin in gastric cancer patients were significantly higher than in controls. The gene expression of bcl-2 was detected in all (100%) and that of proapoptotic bax only in 56% of gastric cancer samples. In comparison to the surrounding non-tumorous tisssue, the gene expression of bax was significantly down-regulated and the gene expression of bcl-2 was up-regulated in gastric cancer tissue. At the protein level, Bax was not detectable and Bcl-2 was seen in 80% of gastric cancer samples. CONCLUSIONS: It is concluded that the patients infected with H. pylori, especially with CagA-positive strains, are at a higher risk of developing a gastric cancer. An increased production and release of gastrin, as well as an over-expression of growth factors such as HGF and TGFalpha, might contribute to the gastric carcinogenesis. In addition, a dysregulation of the Bax/Bcl-2 system with significant up-regulation of Bcl-2 is observed in gastric cancer. PMID- 11421875 TI - The Loiano-Monghidoro population-based study of Helicobacter pylori infection: prevalence by 13C-urea breath test and associated factors. AB - OBJECTIVES: As part of a cross-sectional study on cholelithiasis, 1533 out of 1840 residents in Loiano/Monghidoro, a rural area in Northern Italy (792 men, 741 women, age range 28-80 years), agreed to be further evaluated in relation to their Helicobacter pylori status. METHODS: Each participant performed a 13C-urea breath test (13C-UBT) and provided information on sociodemographic, lifestyle and clinical characteristics. RESULTS: The 13C-UBT was positive in 1041 subjects (67.9%; men: 69%; women: 67%) and was positively associated with increasing age (P < 0.001), alcohol consumption (P < 0.01), a higher number of siblings (P < 0.001) and a personal history of peptic ulcer (P < 0.01), but inversely with a nonmanual occupation (P < 0.001). Overall, H. pylori infection was unrelated to smoking, house pets, and a family history of gastroduodenal diseases. The prevalence of H. pylori infection was 72% in subjects reporting one or more dyspeptic symptoms and 65% among asymptomatic participants (P < 0.001); a multivariate analysis showed that only epigastric pain was significantly, although weakly, associated with 13C-UBT positivity. CONCLUSIONS: This large population-based study showed a prevalence of H. pylori infection higher than that reported by serologic surveys in urban areas. Current H. pylori infection was strongly associated with indicators of lower socioeconomic status, alcohol consumption and increasing age. A role of H. pylori infection in determining epigastric pain was suggested. PMID- 11421876 TI - Economic analysis of four triple regimens for the treatment of Helicobacter pylori-related peptic ulcer disease in in-patient and out-patient settings in Hong Kong. AB - BACKGROUND: One-week triple regimens have been shown to be effective for the treatment of Helicobacter pylori-related peptic ulcer disease. AIM: To conduct an economic analysis of four triple regimens for the treatment of H. pylori-related peptic ulcer disease from the perspective of a public health organization in Hong Kong. METHODS: Records of 200 patients with H. pylori-related peptic ulcer disease, who had previously participated in a randomized comparison of ranitidine bismuth citrate (RBC) with amoxicillin and clarithromycin (RAC), omeprazole with amoxicillin and clarithromycin (OAC), RBC with metronidazole and tetracycline (RMT), or, colloidal bismuth subcitrate with metronidazole and tetracycline (BMT) in either in-patient or out-patient setting were reviewed. RESULTS: Fifty patients were excluded because of incomplete documentation or lack of peptic ulcer. In the out-patient group (n=72), the median direct costs of the RAC group (HK $ 5094) were lower those of the BMT (HK $ 5400), RMT (HK $ 5394), or OAC (HK $ 5440) groups, but the difference was significant only between the RAC and BMT groups (P < 0.05). In the in-patient group (n=78), the median direct costs of the RMT group (HK $ 8524) were significantly lower than those of the OAC (HK $ 13 871) and RAC (HK $ 12 092) groups (P < 0.05), but were similar to those of the BMT group (HK $ 8758). CONCLUSIONS: RAC and RMT are the least costly regimens for out-patient and in-patient treatment, respectively, of H. pylori-related peptic ulcer disease in Hong Kong. PMID- 11421877 TI - Ranitidine bismuth citrate-based triple therapies after failure of the standard 'Maastricht triple therapy': a promising alternative to the quadruple therapy? AB - BACKGROUND: Triple therapy with proton pump inhibitor, clarythromycin, and amoxicillin has been proposed in Maastricht as the first-line treatment of H. pylori infection. AIM: To determine whether ranitidine bismuth citrate (RBC) based regimens may be used as second-line treatments after 'Maastricht therapy' failure. METHODS: A total of 285 patients with H. pylori infection were given a 7 day treatment with pantoprazole 40 mg b.d., clarythromycin 500 mg b.d., and amoxicillin 1 g b.d. Patients who were still infected were randomly given one of the following 14-day treatments: RBC 400 mg b.d. plus amoxicillin 1 g b.d. and tinidazole 500 mg b.d. (RAT group), RBC 400 mg b.d. plus amoxicillin 1 g b.d. and clarythromycin 500 mg b.d. (RAC group), and RBC 400 mg b.d. plus clarythromycin 500 mg b.d. and tinidazole 500 mg b.d. (RCT group). RESULTS: The 'Maastricht therapy' achieved an eradication rate of 59% (95% CI: 54-65) on intention-to treat analysis. The RAT, RAC, and RCT regimens achieved eradication rates of 81% (95% CI: 67-94), 43% (95% CI: 26-60), and 62% (95% CI: 44-80), respectively, on intention-to-treat analysis. Patient compliance was optimal in RAT and RAC groups. CONCLUSION: RBC plus tinidazole and either amoxicillin or clarythromycin can be used as second-line therapies after failure of the Maastricht triple therapy. PMID- 11421878 TI - Predictors of failure of Helicobacter pylori eradication with the standard 'Maastricht triple therapy'. AB - BACKGROUND: Triple therapy with proton pump inhibitor, clarithromycin and amoxicillin has recently been proposed in Maastricht as first-line treatment for H. pylori infection. AIM: To determine predictors of unsuccessful eradication. METHODS: Two hundred and forty-eight patients underwent endoscopy with biopsies for rapid urease test, histology and culture with antibiotic susceptibility tests, and 13C-UBT. All infected patients were given pantoprazole (40 mg b.d.), clarithromycin (500 mg b.d.) and amoxicillin (1 g b.d.) for 1 week. Eradication was assessed by UBT at 4-6 weeks after therapy. RESULTS: One hundred and sixty two of 248 patients (65%) were infected. Culture was positive in 144 (89%). Prevalence rates of metronidazole, clarithromycin and amoxicillin resistance were 14, 8 and 3%, respectively. Eradication rates (95% CI) were 63% (54.7-70.6) by intention-to-treat analysis and 67% (59.4-75.4) by per protocol analysis. Drug compliance was excellent and side-effects were mild. Age > or = 45 years (OR: 2.35, CI: 1.30-4.25), smoking (OR: 1.37, CI 1.01-1.87) and high pre-treatment UBT results (OR: 1.36, CI: 1.08-1.72) were independent predictors of eradication failure. Gender, endoscopic findings, alcohol intake, and clarithromycin and amoxicillin resistance did not predict treatment failure. CONCLUSION: Despite the low prevalence of primary antibiotic resistance in our geographical area, triple therapy with pantoprazole, amoxicillin and clarithromycin achieves low eradication rates. Smoking, age and pre-treatment UBT results are predictors of potential eradication failure. PMID- 11421879 TI - Cure of Helicobacter pylori infection in elderly patients: comparison of low versus high doses of clarithromycin in combination with amoxicillin and pantoprazole. AB - BACKGROUND: Advancing age may influence clarithromycin's pharmacokinetics. No studies have yet compared the effects of different dosages of clarithromycin in combination with a proton pump inhibitor and amoxicillin in elderly patients. AIM: To compare the efficacy and tolerability of clarithromycin 250 mg vs. clarithromycin 500 mg twice daily (b.d.) in combination with pantoprazole and amoxicillin in elderly patients. METHODS: One hundred and fifty-four elderly patients with H. pylori-associated ulcer disease or chronic gastritis were consecutively randomized to receive pantoprazole 40 mg daily plus amoxicillin 1 g, and either clarithromycin 250 mg b.d. (PAC 250) or clarithromycin 500 mg b.d. (PAC 500). Two months after therapy, endoscopy and gastric biopsies were repeated. RESULTS: The cure rates of H. pylori infection in the PAC 250 and PAC 500 groups were, respectively, 83% and 79% (ITT analysis) and 94% and 88% (PP analysis) (P=N.S.). Significant decreases in chronic gastritis activity both in the body (P < 0.00001) and the antrum (P < 0.0001) of the stomach were found in H. pylori-cured patients, independently of clarithromycin dosage. Four patients in PAC 250 (5%) and seven in PAC 500 (9%) reported adverse events (P=N.S.). One patient in PAC 250 (25%) and three in PAC 500 (43%) discontinued the study because of these drug-related side-effects (P=N.S.). CONCLUSIONS: In elderly patients, 1-week triple therapy with a proton pump inhibitor, amoxicillin and clarithromycin is a highly effective and well tolerated anti-H. pylori treatment. With this combination, clarithromycin at the lower dose of 250 mg b.d. achieved excel- lent cure rates and minimized adverse events and costs. PMID- 11421880 TI - Is a one-week course of triple anti-Helicobacter pylori therapy sufficient to control active duodenal ulcer? AB - BACKGROUND: Triple therapy currently forms the cornerstone of the treatment of patients with Helicobacter pylori-positive duodenal ulcer. AIM: To establish whether prolonged antisecretory therapy is necessary in patients with active duodenal ulcer. METHODS: A total of 77 patients with H. pylori-positive duodenal ulcer were included in a prospective, controlled, double-blind study. All patients received a 7-day treatment with omeprazole 20 mg b.d., clarithromycin 500 mg b.d. and amoxicillin 1000 mg b.d. Patients in the omeprazole group underwent an additional 14-day therapy with omeprazole 20 mg; patients in placebo group received placebo. Endoscopy was performed upon inclusion in the study and after 3 and 8 weeks. RESULTS: Seventy-four patients were eligible for a per protocol analysis after 3 weeks, and 65 after 8 weeks. After 3 weeks, the healing rate was 89% in the omeprazole group and 81% in the placebo group (P=0.51). After 8 weeks, the ulcer healed in 97% of the patients in the total group (95% CI: 92.7 100%). H. pylori was eradicated in 88% of patients in the omeprazole group and in 91% in the placebo group (P=1.0). No statistically significant differences between the groups were found in ulcer-related symptoms or in ulcer healing. CONCLUSION: In patients with H. pylori-positive duodenal ulcer, a 7-day triple therapy alone is sufficient to control the disease. PMID- 11421881 TI - Arrest of chronic acid suppressant drug use after successful Helicobacter pylori eradication in patients with peptic ulcer disease: a six-month follow-up study. AB - BACKGROUND: It remains controversial whether successful H. pylori eradication leads to relief of dyspepsia and the subsequent arrest or tapering of acid suppressant drug therapy, or to an aggravation of acid-related dyspepsia requiring more acid-suppressant drug intake. AIM: To evaluate prospectively the effect of H. pylori eradication on the requirement of acid-suppressant drug or antacids and the evolution of dyspeptic symptoms in chronic acid-suppressant drug users with peptic ulcer disease. MATERIALS AND METHODS: The use of acid suppressant drugs, rescue antacids and predominant symptoms were recorded prospectively during 24 weeks after H. pylori eradication therapy in 75 peptic ulcer disease patients. RESULTS: In 71 patients with complete follow-up, ulcers were healed at follow-up endoscopy and H. pylori was successfully eradicated. After 6 months, 93% (66 out of 71) of chronic acid-suppressant drug users had stopped acid-suppressant drug intake. The mean daily acid-suppressant drug dosage per patient decreased from 1.72 at entry to 0.03 units acid-suppressant drug (98%; P < 0.0001) during follow-up. The mean number of antacid tablets/day/patient was 0.26 during follow-up for the relief of mild inter current dyspeptic symptoms. Medication use was not different in peptic ulcer disease patients with or without gastro-oesophageal reflux disease at baseline. The prevalence of gastro-oesophageal reflux disease decreased from 42% before to 35% after H. pylori eradication (N.S.). CONCLUSION: Successful H. pylori eradication in peptic ulcer disease patients almost completely eliminates the need for acid-suppressant drug regardless of the presence or absence of gastro oesophageal reflux disease at entry. PMID- 11421882 TI - Helicobacter pylori infection as a risk factor for gastrointestinal symptoms in patients using aspirin to prevent ischaemic heart disease. AB - BACKGROUND: Aspirin use in the secondary prevention of ischaemic heart disease may provoke gastrointestinal discomfort. OBJECTIVE: To register gastrointestinal symptoms and complications in patients with cardiovascular disease using aspirin and to relate these symptoms to infection with H. pylori. METHODS: Blood samples were obtained from 398 consecutive patients in the Coronary-Care Unit, University Hospital Nijmegen and analysed for serum antibody levels to H. pylori infection. Questionnaires were sent 2 weeks after discharge to assess gastrointestinal symptoms. RESULTS: Questionnaires were returned by 314 patients (79%). A total of 183 out of 314 patients (46%) reported gastrointestinal symptoms. Of 238 patients using 80-100 mg aspirin daily, 145 (61%) recorded gastrointestinal symptoms. Besides aspirin, the use of calcium antagonists was correlated with gastrointestinal symptoms. Of the 128 patients using calcium antagonists, 84 (66%) reported gastrointestinal symptoms. The prevalence of gastrointestinal symptoms in H. pylori-positive and -negative patients using aspirin was 48% and 52%, respectively. CONCLUSIONS: Two weeks after discharge almost 50% of the patients with cardiovascular disease experienced gastrointestinal symptoms, especially patients using aspirin or calcium antagonists. Patients seropositive for H. pylori and using aspirin or calcium antagonists did not have more gastrointestinal discomfort compared to non-infected patients. PMID- 11421883 TI - Modified seven-day, quadruple therapy as a first line Helicobacter pylori treatment. AB - BACKGROUND: Cure rates of 7-day triple therapy seem to be decreasing. Quadruple therapies may be an alternative, although their complex administration makes patient acceptance difficult. OBJECTIVE: To test the usefulness of a thrice a day, quadruple therapy to cure Helicobacter pylori infection. PATIENTS AND METHODS: A total of 122 consecutive patients with peptic ulcer and Helicobacter pylori infection were treated with omeprazole 20 mg b.d., tetracycline chlorhydrate 500 mg t.d.s., metronidazole 500 mg t.d.s., and bismuth subcitrate 120 mg t.d.s. administered with meals for 7 days. Cure was tested by either endoscopy or breath test after 2 months, and by urea breath test 6 months after therapy. RESULTS: Seven patients were lost to follow-up. Of the remaining 115, 110 were cured at the first control, giving an intention-to-treat cure rate of 90.2% (95% CI: 83-95%) and a per protocol cure rate of 95.7% (95% CI: 90-98%). One hundred three patients returned for a 6-month breath test; all but one were cured. Side-effects were minimal or minor in 47 patients (40.8%) and moderate in four (3.4%). Compliance was good, 95% of patients taking more than 90% of the pills. Six (5%) patients stopped treatment after 1, 2, 4 (two patients) and 6 (two patients) days. CONCLUSION: Thrice a day quadruple therapy shows excellent cure rates, far above 90%, is well-tolerated and compliance is easy. Head-to-head comparison with triple therapies as first line Helicobacter pylori treatment seems warranted. PMID- 11421884 TI - What is the optimal length of proton pump inhibitor-based triple therapies for H. pylori? A cost-effectiveness analysis. AB - BACKGROUND: Triple therapy with a proton pump inhibitor, clarithromycin and amoxicillin is widely used for H. pylori infection. The appropriate length of treatment remains controversial. AIM: To determine whether length of treatment has an impact on the cost-effectiveness of triple therapy. METHODS: The study took the form of a cost-effectiveness analysis spanning 2 years. The perspective was societal and the setting, ambulatory care. Subjects were Helicobacter pylori positive patients with a duodenal ulcer. The triple therapy trials spanned 7, 10 or 14 days and the main outcome measures were cost per patient and marginal cost for additional cured patient calculated for a low cost-of-care setting (Spain), for a high-cost setting (USA), and for two follow-up strategies: (i) systematic 13C-urea breath test after eradication; (ii) clinical follow-up, breath-test if symptoms recurred. RESULTS: Base-case analysis showed that for both the 13C-UBT and the clinical follow-up branches, lowest costs were obtained with 7-day schedules both in Spain and the USA. Sensitivity analysis showed that in Spain, 10-day therapies would have to increase 7-day cure rates by 10-12% to become cost effective. In contrast, in the USA only a 3-5% increase was needed. The corresponding figures for 14-day therapy were 25-35% and 8-11%, respectively. CONCLUSIONS: Seven-day therapies seem the most cost-effective strategy. However, in high-cost areas the differences were less evident. PMID- 11421885 TI - Comparison of the effects of concanavalin-A and epidermal growth factor on epithelial cell proliferation in the rat intestine. AB - BACKGROUND: Concanavalin-A, the lectin present in Jack beans, binds to mannose- and glucose-containing residues and can interact with the epidermal growth factor receptor and moderate cell proliferation in vitro. AIM: To compare the actions of concanavalin-A and epidermal growth factor on the gastrointestinal tract in vivo. METHODS: Rats maintained on total parenteral nutrition were given intragastric concanavalin-A, intravenous epidermal growth factor or concanavalin-A and epidermal growth factor. Cell proliferation and crypt fission were assayed in 'micro-dissected' crypts. RESULTS: Concanavalin-A and epidermal growth factor both significantly elevated proliferation in the small intestine and colon. No significant interaction between the effects of these two agents was seen, except in the mid small intestine where there was a synergistic interaction. Concanavalin-A had no effect on crypt branching. Epidermal growth factor significantly reduced branching in the distal small intestine and mid colon. CONCLUSION: The effects of the two agents appeared to be separate, except in the mid small intestine where they were additive. This is in marked contrast with the actions reported in vitro, where concanavalin-A is a powerful inhibitor of epidermal growth factor-induced cell proliferation. Concanavalin-A thus has potential for enhancing the functions of the small intestine. PMID- 11421886 TI - Potential gastrointestinal effects of long-term acid suppression with proton pump inhibitors. PMID- 11421889 TI - Exposure to environmental tobacco smoke and adult asthma. PMID- 11421890 TI - Immunity, hormones, and the brain. PMID- 11421891 TI - Cross-reactivity of IgE antibodies to allergens. AB - The cross-reactivity of IgE antibodies is of interest for various reasons, three of which are discussed. Firstly, from the clinical view, it is important to know the patterns of cross-reactivity, because they often (but not always) reflect the pattern of clinical sensitivities. We discuss the cross-reactivities associated with sensitization to pollen and vegetable foods: PR-10 (Bet v 1-related), profilin, the cross-reactive carbohydrate determinant (CCD), the recently described isoflavone reductase, and the (still elusive) mugwort allergen that is associated with celery anaphylaxis; cross-reactivities between allergens from invertebrates, particularly tropomyosin, paramyosin, and glutathione S transferase (GST); and latex-associated cross-reactivities. Clustering cross reactive allergens may simplify diagnostic procedures and therapeutic regimens. Secondly, IgE cross-reactivity is of interest for its immunologic basis, particularly in relation to the regulation of allergic sensitization: are IgE antibodies to allergens more often cross-reactive than IgG antibodies to "normal" antigens? If so, why? For this discussion, it is relevant to compare not only the structural relation between the two allergens in question, but also the relatedness to the human equivalent (if any) and how the latter influences the immune repertoire. Thirdly, prediction of IgE cross-reactivity is of interest in relation to allergic reactivity to novel foods. Cross-reactivity is a property defined by individual antibodies to individual allergens. Quantitative information (including relative affinity) is required on cross-reactivity in the allergic population and with specific allergens (rather than with whole extracts). Such information is still scarce, but with the increasing availability of purified (usually recombinant) allergens, such quantitative information will soon start to accumulate. It is expected that similarity in short stretches of the linear amino-acid sequence is unlikely to result in relevant cross-reactivity between two proteins unless there is similarity in the protein fold. PMID- 11421892 TI - In utero and perinatal complications preceding asthma. AB - BACKGROUND: It has been suggested that pregnancy and early life may influence the development of asthma in the offspring, but published studies have not carefully controlled for potential biases. METHODS: In a large British birth cohort of 4065 natural children of 2583 mothers, we investigated whether in utero and perinatal influences contribute to the development and the severity of asthma in childhood, allowing for possible confounders of the relationship, and considering the nonindependence of familial data. RESULTS: Child asthma (10.1%) was more frequently reported by mothers when there had been health complications during pregnancy (prevalence =14.3%; adjusted odds ratio [ORadj] =2.01; 95% confidence interval, 1.52-2.67), labor, or delivery (19.3%, ORadj =1.35, 1.01-1.81); child illness or health complications during the first week of life (22.6%, ORadj =1.35, 1.01-1.82); and birth weight of < 2.5 kg (7.0%, ORadj =1.57, 1.10-2.25). Specific causes of health complications during pregnancy which significantly related to asthma were early or threatened labor (ICD: 644) (4.8%, ORadj =1.58, 1.03-2.40) and the malposition or malpresentation of the fetus (ICD: 652) (1.6%, ORadj =3.63, 1.47-8.91). CONCLUSION: The results provide further evidence that in utero and perinatal factors may increase the risk of developing asthma. PMID- 11421893 TI - A well-tolerated grass pollen-specific allergy vaccine containing a novel adjuvant, monophosphoryl lipid A, reduces allergic symptoms after only four preseasonal injections. AB - BACKGROUND: We present data showing that a Th1-inducing adjuvant can reduce the number of injections required for allergy vaccination. Allergy vaccination is the only treatment for type 1 hypersensitivity that can alter the underlying disease process. A switch of specific T-cell activity from Th2 >Th1 to Th1 >Th2 is believed to be an important change seen after long-term vaccination therapy. An immunologic adjuvant that enhances such a switch could be used to reduce the number of injections required. This would improve compliance with the treatment and provide pharmacoeconomic advantages. Such an adjuvant is 3-deacylated monophosphoryl lipid A (MPL adjuvant, Corixa). METHODS: A multicentre, placebo controlled, randomized, double-blind clinical study was performed with a new standardized allergy vaccine comprising a tyrosine-adsorbed glutaraldehyde modified grass pollen extract containing MPL adjuvant. Four subcutaneous injections of the active product were given preseasonally to 81 grass pollen sensitive subjects, and 60 received placebo injections (tyrosine alone). Diary cards were used to record symptoms and medication taken during approximately 30 days of the grass pollen season. RESULTS: There was a statistical advantage in favour of the active treatment for nasal (P = 0.016) and ocular (P = 0.003) symptoms and combined symptom and medication scores (P=0.013). Titrated skin prick testing revealed a significant reduction of skin sensitivity in the active group compared to placebo (P = 0.04). Grass-pollen-specific IgG antibody was raised by active treatment (P < 0.01). A rise in IgE antibody was seen in the placebo group during the season (P < 0.01). The first year's treatment rise of IgE was not seen in the active group, and no rise occurred during the pollen season. More local adverse events were seen in the active group. There was no difference in generalized adverse events. CONCLUSION: A new, well-tolerated allergy vaccine, incorporating a Th1-inducing adjuvant, MPL, was efficacious and after only four preseasonal injections produced antibody changes normally associated with long injection schedules. This may encourage wider application of allergy vaccination. The vaccine is now available in a number of countries as Pollinex Quattro. PMID- 11421894 TI - A prospective study of the relationship between exposure and specific antibodies in workers exposed to organic acid anhydrides. AB - BACKGROUND: The exposure-response relationships for the induction of specific IgE and IgG were evaluated in a prospective study of workers exposed to organic acid anhydrides (OAAs). Special attention was paid to the modifying effects of atopy and smoking. METHODS: The subjects were 163 previously unexposed persons exposed to epoxy resins with hexahydro-, methylhexahydro-, and methyltetrahydrophthalic anhydride as curing agents. The levels of OAAs in air and of specific IgE and IgG in serum were recurrently monitored. The mean observation time was 32 (1-105) months. RESULTS: The mean combined OAA exposure of the subjects was 15.4 (< 1 189) microg/m3. Positive specific IgE was demonstrated by 21 (13%) subjects with a mean induction time of 8.8 (1-35) months. The incidence of sensitization was 4.1 cases/1000 months at risk. The relative risk (OR) for atopics was 5.4 (1.9 15.3; 95% CI). An exposure-response relationship was demonstrated by an increasing risk of sensitization with increasing exposure. CONCLUSION: An association between exposure and atopy, respectively, and the induction of specific antibodies against OAAs were observed. The risk for atopics was comparable with the risk for the subjects in the most exposed group. PMID- 11421895 TI - Systemic ketoconazole is an effective treatment of atopic dermatitis with IgE mediated hypersensitivity to yeasts. AB - BACKGROUND: IgE-mediated hypersensitivity to yeasts is often seen in atopic dermatitis (AD) patients, especially when dermatitis is located in the head, neck, and shoulder regions. Two studies have shown the efficacy of ketoconazole in the treatment of this type of AD, in contrast to results of topical treatment. The objective was to assess the clinical efficacy of antifungal treatment in AD in a randomized, double-blind, placebo-controlled study with oral ketoconazole and yeast-specific IgE levels and saprophytic yeast growth monitored simultaneously. METHODS: Eighty patients with AD and positive P. ovale and/or C. albicans RAST/skin prick test results were randomized to receive ketoconazole or placebo for 30 days. The yeast growth of skin and pharynx; P. ovale, C. albicans, andS. cerevisiae RAST; serum total IgE; and the severity of the eczema (SCORAD) were assessed at day 0 and thereafter at 1 and 3 months. RESULTS: A significant improvement was seen in the SCORAD scale in the ketoconazole group at the second visit in comparison to the first visit (P<0.0005; n=36), but not in the placebo group (n=39). Of the individual determinants of the SCORAD, itching (P<0.005), the extent of dermatitis (area percentage), excoriation, lichenification (P<0.01), erythema, papulation, and dryness (P<0.05) improved significantly in the ketoconazole group. In the placebo group, only the extent of dermatitis (area percentage) decreased significantly (P<0.05). In the ketoconazole group, the number of positive P. ovale cultures decreased from 60% to 31% (n=35) compared to the placebo group (64% to 56%; n=39). The clinical response was most significant in female patients with positive yeast cultures. CONCLUSION: Saprophytic yeasts may be a source of allergens in AD. Thus, patients with AD, yeast growth, and elevated IgE levels to yeasts should be offered antifungal treatment. PMID- 11421896 TI - Double-blind, randomized, placebo-controlled trial of effect of nedocromil sodium on clinical and inflammatory parameters of asthma in children allergic to dust mite. AB - BACKGROUND: The purpose of this study was to determine the clinical effect of nedocromil sodium and its relationship with serum levels of inflammatory mediators by monitoring lung function and noninvasive markers of airway inflammation, such as eosinophil blood counts; serum ECP, sIL-2R, IL-4 and sICAM; and total IgE. Anti-inflammatory medications cause a reduction in the markers of airway inflammation, decrease the intensity of airway hyperresponsiveness, and improve clinical symptoms of asthma. Among the available choices is nedocromil sodium, which is favored in the treatment of asthmatic children due to its very mild side-effects. It has been previously shown to improve the clinical parameters of asthma, but there are limited data on its effect on inflammatory mediators in the serum of asthmatic children. METHODS: In this double-blind, randomized, placebo-controlled 8-week trial, 39 children, aged 9-16 years, with moderate atopic asthma were randomly allocated to receive either nedocromil sodium, two puffs twice daily, 0.002 g/puff, or placebo, two puffs twice daily. The primary end points were the clinical parameters of asthma measured by asthma symptom score, FEV1, and PC20H. Other end points included the serum levels of various inflammatory markers - ECP, sIL-2R, IL-4, sICAM, and IgE. RESULTS: Clinical and inflammatory parameters improved with the use of nedocromil sodium, compared with placebo. Nedocromil significantly decreased serum levels of inflammatory markers, as shown in the following table. No correlation was found between any of the measured parameters. CONCLUSION: Nedocromil sodium provided effective anti-inflammatory treatment for children with moderate atopic asthma. PMID- 11421897 TI - Sex-related differences in the initiation of allergic rhinitis in mice. AB - BACKGROUND: Several clinical and epidemiologic studies have investigated sex differences in the prevalence of allergic rhinitis. At present, however, no reports have demonstrated such differences in experimental models with local, but not parenteral, sensitization with antigens that may reflect natural exposure to allergens. We have recently developed murine models of allergic rhinitis after repeated intranasal sensitization with antigens in the absence of adjuvants. In this study, we investigated the role of sex in the initiation of the disease in vivo. METHODS: Male and female CBA/J and BALB/c mice were sensitized intranasally with phospholipase A2 (PLA2) and Schistosoma mansoni egg antigen (SEA), respectively, in the absence of adjuvants. After the repeated sensitization, serum Ab titers against the sensitizing antigen and nasal eosinophilia were determined. In addition, the involvement of androgen in IgE synthesis was investigated in castrated CBA/J male mice with or without testosterone administration. RESULTS: Females produced significantly higher levels of PLA2 specific IgE than males in CBA/J mice sensitized with PLA2. On the other hand, both titers of PLA2-specific IgG1 and nasal eosinophilia did not significantly differ between the two groups. Castrated male mice produced significantly higher amounts of PLA2-specific IgE than sham-treated male mice. In addition, PLA2 specific IgE production decreased in castrated mice treated with testosterone. Sexual differences in the production of Ag-specific IgE were not seen in BALB/c mice after the sensitization with SEA. CONCLUSION: These results suggest that sex is responsible for the production of Ag-specific IgE, but not IgG1 or nasal eosinophilia, and that androgen appears to be involved in the in vivo production of specific IgE in male mice. PMID- 11421898 TI - Presence of fluticasone propionate on human nasal mucosal surface and in human nasal tissue over a period of 24 h after intranasal application. AB - BACKGROUND: Once-daily use of nasally applied glucocorticoids was demonstrated to be effective in the treatment of allergic rhinitis. The aim of the study was to measure concentrations of fluticasone propionate (FP) in nasal secretion and nasal tissue over a period of 1 day after a single application of 100 microg FP. METHODS: Twenty-six patients applied nasal FP spray at different time intervals before surgery. Cotton swabs, used to clean the mucosal surface, and resected nasal tissue were extracted. FP concentrations were determined by RIA. RESULTS: FP was found in nasal secretions in concentrations from 15 to 1 microg/g over a period of 20 h, and in nasal tissue in concentrations from 200 to 13 ng/g up to 24 h after the single application. CONCLUSION: The long-persisting high concentrations of FP provide the pharmacokinetic basis for once-daily treatment. PMID- 11421899 TI - Specific IgE and IgG to gelatin in children with systemic cutaneous reactions to Japanese encephalitis vaccines. AB - BACKGROUND: Systemic allergic reactions to Japanese encephalitis (JE) vaccine that include urticaria, angioedema, and rash have been reported. In Japan, children who suffered from allergic immediate-type reactions to JE vaccine had antigelatin IgE in their sera. However, the immunologic mechanism of allergic nonimmediate-type reactions that consist of cutaneous signs appearing several hours or more after JE vaccination has not been defined. METHODS: Serum samples were taken from 28 children who showed allergic nonimmediate-type cutaneous reactions to JE vaccine. Furthermore, serum samples were taken from 10 children who showed allergic immediate-type reactions with cutaneous signs and/or respiratory symptoms to JE vaccine. We have defined an immediate-type reaction as one occurring within 1 h after vaccination. RESULTS: Of 10 children who showed immediate-type reactions, all had antigelatin IgE and IgG. Of 28 children who showed systemic nonimmediate-type reactions, one had antigelatin IgE and nine (32%) had antigelatin IgG. The child who had antigelatin IgE showed urticaria 2 h after JE vaccination. CONCLUSION: These results suggest that some children who showed allergic nonimmediate-type reactions to JE vaccine were sensitized to gelatin. PMID- 11421900 TI - An Internet-based survey of anaphylactic reactions to foods. AB - BACKGROUND: A favorable outcome of anaphylaxis depends on the rapidity of adequate initial management and epinephrine injection. However, few data on the management of anaphylactic reactions are available. The aim of this study was to investigate the management and awareness of anaphylaxis to foods by mean of an Internet-based survey. METHODS: Visitors to a website with information on food allergy were invited to join the survey. Items in the survey included the management of anaphylactic reactions, investigations done by the diagnosing physician, and information given to the responder in anticipation of a new anaphylactic reaction. RESULTS: Almost all of the 264 responders were from North America, Europe, Australia, or New Zealand (263/264). The most recent reaction was treated by epinephrine injection in 68.7% (180/262) of cases, or by antihistamines in 14.1% (37/262). Epinephrine was the first treatment for the most severe reaction in only 43.9% (101/230), while antihistamines were given first in 43.5% (100/230). One-third (62/210 = 29.5%) of the responders diagnosed by a physician received neither a diagnostic blood test nor a skin test. Responders diagnosed by an allergist were more often investigated (91/105 = 86.7%) than those diagnosed by a pediatrician or an internist (29/44 = 65.9%), a general practitioner (22/45 = 48.9%), or another physician (6/16 = 37.5%) (P < 0.001). Most responders had received instructions on how to deal with a new episode of anaphylaxis (244/263 = 92.8%). Responders instructed by an allergist were most frequently satisfied with the instructions (115/131 = 87.8%). CONCLUSION: A large number of responders did not receive epinephrine for treatment of their most severe, or most recent anaphylactic reaction, and did not undergo allergy tests. The conventional management of anaphylaxis might still be improved. PMID- 11421901 TI - Comparison of ELISA for tissue transglutaminase autoantibodies with antiendomysium antibodies in pediatric and adult patients with celiac disease. AB - BACKGROUND: Tissue transglutaminase (t-TG) is the main autoantigen recognized by the endomysium antibodies (EMA) observed in patients with celiac disease (CD). The aim of the study was to assess an ELISA method for t-TG antibodies (t-TGA) with respect to EMA IF assay in pediatric and adult patients. METHODS: t-TGA were analyzed by ELISA in 220 sera samples: 82 patients with biopsy-proven untreated CD (23 adults and 59 children), 14 CD children on gluten-free diet, 18 asymptomatic relatives of CD patients, and 106 age-matched control patients with gluten-unrelated gastrointestinal diseases (58 adults and 48 children). Serum IgA EMA were tested on umbilical cord sections in all patients. RESULTS: The great majority (92.7%) of untreated CD patients (both adults and children) were t-TGA positive (values ranging from 20.1 to > 300 AU). None of the child control patients and only two out of 58 (3.4%) of the adults with unrelated gastrointestinal diseases had serum t-TGA positivity; two out of 18 first-degree relatives with biopsy-proved silent CD were t-TGA (as well as EMA) positive. Finally, two out of 14 CD children, assuming a gluten-free diet, had serum t-TGA (as well as EMA). A highly significant correlation (P < 0.001) was observed between t-TGA concentrations and EMA. t-TGA showed a sensitivity of 87% and 95%, a specificity of 97% and 100% for adults and children, respectively. CONCLUSION: The method is highly sensitive and specific in the diagnosis of CD and is promising as a tool for routine diagnostic use and population screening, especially in children. PMID- 11421902 TI - Diagnostic evaluation of grass- and birch-allergic patients with oral allergy syndrome. AB - BACKGROUND: Patients with birch and grass allergy often suffer from oral allergy symptoms when ingesting cross-reacting fresh fruits and vegetables. However, fruit and vegetable allergen extracts are often readily degradable or contain clinically irrelevant cross-reacting epitopes, resulting in diagnostic discrepancies when fruit and vegetable allergic reactions are evaluated. The risk of using nonstandardized fresh food extracts for skin testing may also be of concern. The objective was to compare and evaluate the clinical utility of selected recombinant grass and birch cross-reacting food allergens with fresh and commercial melon, hazelnut, and apple extracts. METHODS: Thirty-six grass- and or birch-allergic patients and 17 control subjects consented to participate in the study. All subjects were skin prick tested and had basophil histamine-release tests done with fresh fruits and various extracts of hazelnut, apple, and melon. The diagnosis of oral allergy syndrome was confirmed by oral challenges. In addition, histamine release to recombinant Bet v 1 and Bet v 2, and recombinant Phl p 1 and Phl p 2, Phl p 5 was performed. RESULTS: The skin prick test with fresh hazelnut, apple, and melon showed sensitivities of 0.97, 0.92, and 0.89, respectively. The corresponding specificities were 0.78, 0.72, and 0.82, respectively. In contrast, the histamine-release test with hazelnut, apple, and melon gave sensitivities of 0.87, 0.71, and 1.00, respectively. The corresponding specificities were 0.65, 0.93, and 0.43. The skin prick test showed excellent negative predictive value (> 90%). No added value of recombinant allergen testing was noted. Oral challenge did not result in severe systemic reactions, and no systemic reactions were observed with skin prick tests with fresh fruits. CONCLUSION: The skin prick test showed an almost optimal diagnostic value with a satisfactory sensitivity (> 89%) and excellent negative predictive value with fresh fruits. When the skin prick test with fresh nut and apple cannot be performed, histamine release is a diagnostic alternative. Histamine release with melon showed lack of specificity. This was probably due to extensive IgE cross reactivity with pollen, since these patients also responded to recombinant Phl p 1 and Bet v 1. Skin testing and challenges with fresh fruits were safe. PMID- 11421903 TI - Duration of the antihistaminic effect after discontinuation of ebastine. AB - BACKGROUND: The inhibitory effect of antihistamines on allergen-induced skin reactions can impair the results of allergen skin testing, which are necessary for the diagnosis of atopic diseases. This study was designed to determine the time period required for the inhibitory effect of ebastine on allergen-induced skin reactivity to disappear completely. METHODS: This was a double-blind, placebo-controlled, parallel-group study including 23 out of 27 randomized patients. They received either ebastine 20 mg or placebo once daily for 7 days. At the end of treatment, allergen challenge was performed daily for 7 days. Histamine challenge was performed on day 1 (6 and 24 h) and day 5 after treatment. The wheal and flare surface areas were measured and analyzed. RESULTS: Highly significant inhibition of the wheal and flare response induced by allergen was observed after ebastine treatment on days 1 and 2 as compared with placebo (P < 0.01 for both). The inhibition was reduced, although still significant, by day 3 (P < 0.05). No significant difference was observed by day 4 between the ebastine and the placebo groups. The effects of histamine challenge were significantly reduced in the ebastine compared with the placebo group at day 1 (6 and 24 h), and were similar at day 5 after treatment. CONCLUSION: Our results show that the wheal and flare response to allergen after ebastine discontinuation returns to placebo values after 4 days. Therefore, patients using ebastine need to be antihistamine-free for 4 days before the skin prick test. This is valuable information for the allergologist seeking to diagnose allergen sensitivity. PMID- 11421904 TI - Trichophyton sensitivity in allergic and nonallergic asthma. AB - BACKGROUND: Although the role of inhaled fungi in inducing asthma has been repeatedly confirmed, there are few reports about the association of asthma with dermatophyte sensitivity and the causal role of Trichophyton allergy in asthma. The objective was to investigate the presence of Trichophyton sensitivity among patients with allergic and nonallergic asthma in combination with tinea, and to compare the situation with several control groups in order to evaluate the factors determining Trichophyton sensitivity. METHODS: A total of 86 subjects (55 female, 31 male) with a mean age of 38.6 +/- 11.1 years were included in the study. The patients were divided into five groups: 1) nonallergic asthma plus tinea (n = 19) 2) allergic asthma plus tinea (n = 15) 3) asthma without tinea (n = 22) 4) tinea without asthma (n = 17) 5) healthy controls (n = 13). Skin tests with standardized extracts of T. rubrum and specific IgE measurements were performed in all subjects. All patients were also subjected to microscopic evaluation and fungal culture for dermatophyte infection. RESULTS: The skin test positivity rate to Trichophyton extract of groups 1 (63.1%), 2 (46.7%), and 4 (47.1%) was higher than that in groups 3 (4.4%) and 5 (7.7%) (P < 0.05). Although not significant, the rates of sensitivity to T. rubrum (63.1%) and of severe asthma (31.6%) were higher in the group with nonallergic asthma with tinea (group 1) than in other groups. Among 51 patients in whom direct microscopic evaluation revealed dermatophyte infection, 60.8% had positive fungal cultures for T. rubrum (58.1%), T. mentagrophytes (35.5%), and Candida (6.4%). CONCLUSION: According to our data, the presence of fungal infection seems to be an important determinant in hypersensitivity to Trichophyton whether or not the subject is asthmatic and/or allergic. Since a greater proportion of patients with nonallergic asthma- in whom the rate of severe asthma was also higher - showed positive skin tests to Trichophyton extracts in this study, we believe that patients with severe, intrinsic asthma should be examined for signs of fungal infection and tested to determine immediate hypersensitivity to dermatophyte antigens. PMID- 11421905 TI - Analysis of allergens in rat fur and saliva. AB - BACKGROUND: The fur is an important source of allergens in many mammal species, but this source has not been extensively studied in rats. Rat room dust contains high-molecular-weight allergens that have been found to cross-react with fur and its presumed salivary contaminants. The role of rat fur and saliva as a source of respiratory allergens merits further investigation. The objective was to describe the allergens present in the fur and saliva of male rats. METHODS: Allergen extracts were prepared from the saliva and fur of the pelts from adult male rats. Immunoblotting was used to describe the allergens present in rat fur and saliva with serum from 76 and 25 individuals, respectively. RESULTS: There was considerable variation between the individuals in the binding of IgE to the separated fur and saliva allergens. Immunoblot analysis identified 23 allergens in rat fur. "Major" allergens were found at the "origin", and at 55, 51, 19, and 17 kDa, and "intermediate" allergens at 74, 67 (probably albumin), and 21.5 (diffuse) kDa. Seventeen salivary allergens were described with "major" allergens at 21.5, 19.5, 19, 18, and 17.5 kDa. Many subjects had IgE to the 67 kDa (56%) and 43 kDa (64%) allergens but the density of staining was weak. CONCLUSION: Rat fur and saliva are the sources of many allergens. Fur contained five "major" allergens and was a complex source of allergens of relatively high molecular weight (>22 kDa). The most important salivary allergens have molecular weights of less than 22 kDa. Fur is the most probable source of the high-molecular-weight allergens found in rat room dust. PMID- 11421906 TI - Variations of serum eosinophil cationic protein and tryptase, measured in serum and saliva, during the course of immediate allergic reactions to foods. AB - BACKGROUND: Subjective complaints and reactions after placebo administration during food challenges (FC) may make their outcome difficult to interpret. We determined serum ECP and tryptase as tryptase in saliva during FC, looking for markers to support challenge outcomes. METHODS: Twelve patients with systemic reactions after food intake and nine presenting oral allergy syndrome (OAS) underwent skin tests; total and specific IgE determination; double-blind, placebo controlled FC (DBPCFC); and open challenges. Blood samples were collected before and 1, 2, and 5 h after challenge and saliva before and 5, 30, and 60 min after challenge. ECP and tryptase were quantified by ImmunoCAP (Pharmacia-Upjohn, Sweden). Serum tryptase of > 10 microg/l was considered positive. RESULTS: After positive DBPCFC (n = 8), ECP rose significantly (P < 0.05) at 1-h - 16.03 (12.8) microg/l (mean [standard deviation]) - and 2-h intervals - 17.56 (10.7) microg/l compared to basal level of 9 (6.4) microg/l. After negative DBPCFC (n = 6), ECP increased from basal 9.63 (3.9) microg/l to 24.84 (14.17) microg/l at the 2-h time point. There were nonsignificant differences in ECP between patients with positive and negative FC. Two patients with positive challenge showed a tryptase level of >10 microg/l and only one patient with OAS showed 5.6 microg/l of tryptase 5 min after FC. CONCLUSIONS: ECP and tryptase in serum and saliva were not useful markers for FC outcomes. PMID- 11421907 TI - Specific IgE antibodies to peanut in western Sweden--has the occurrence of peanut allergy increased without an increase in consumption? AB - BACKGROUND: Sensitization to peanut has seldom been investigated in Sweden. Therefore, all IgE-specific tests for peanut during a 5-year period were reviewed to study the relation between the levels of specific IgE antibody to peanut and age, sex, symptoms, and other atopic manifestations. METHODS: All serum samples were analyzed for IgE antibodies to peanut in relation to sex, age, clinical reactions, and other food allergens. A subgroup was asked to answer a questionnaire about symptoms and atopic manifestations in relation to IgE antibody levels. RESULTS: During the study period, 2417 tests were made for peanut. There was an increased prevalence of detectable IgE antibodies during the years studied. More than 80 individuals under 2 years of age were sensitized to peanut. In the subgroup, individuals with detectable IgE antibodies reported a shorter reaction time after eating peanuts than individuals with normal IgE antibody levels (P < 0.05). CONCLUSION: The reaction pattern to peanuts in Sweden is similar to that in many other countries despite a reported steady and low consumption. The severity of symptoms was connected to age and IgE antibody level. Patients with normal or low IgE antibody levels were not always free of symptoms even though their risk of allergic symptoms was reduced. PMID- 11421908 TI - Sublingual swallow or spit? PMID- 11421909 TI - Propolis, a new bee-related allergen. PMID- 11421910 TI - IgE-mediated anaphylaxis to antacid. PMID- 11421911 TI - Adverse reactions to sculptured nails. PMID- 11421912 TI - Allergenicity of the nan-gai nut. PMID- 11421913 TI - Airborne allergic contact dermatitis. PMID- 11421914 TI - Ficus antigen in dust. PMID- 11421915 TI - Are human milk polyunsaturated fatty acids (PUFA) related to atopy in the mother and her child? PMID- 11421916 TI - The role of CD30 in atopic disease. PMID- 11421917 TI - Home allergen monitoring and control--improving clinical practice and patient benefits. PMID- 11421918 TI - In vitro T-cell responses to beta-lactam drugs in immediate and nonimmediate allergic reactions. AB - BACKGROUND: beta-Lactam drugs may induce both cellular and humoral allergic reactions, and there is evidence that T cells play an important role in the pathogenesis of these reactions. The aim of this work was to assess the sensitivity and specificity of the lymphocyte transformation test (LTT) as an in vitro diagnostic tool, in patients with either an immediate or a nonimmediate reaction to penicillin G and/or amoxicillin. METHODS: Fifty patients with a well documented history of allergic reactions to beta-lactams (31 immediate and 19 nonimmediate) were studied by means of skin tests (prick and intradermal), radioallergosorbent test (RAST), and, when necessary, controlled administration of the drug. Twenty-eight healthy subjects with good tolerance to penicillins served as controls. LTT was performed in all subjects. RESULTS: Skin tests were positive in 77.4% of the patients with immediate reactions and in 36.8% of those with nonimmediate reactions. The overall sensitivity of LTT in the allergic patients was 62%, but, when analyzed separately, sensitivity was 64.5% for the immediate group and 57.9% for the nonimmediate group. The LTT specificity was 92.8%. CONCLUSION: The LTT should be considered a useful in vitro diagnostic tool to identify subjects allergic to penicillins, especially patients with nonimmediate reactions where the LTT has a better diagnostic value than skin tests. Interestingly, positive T-cell proliferative responses can be observed 10 or more years after the occurrence of the reaction without further exposure to the drug. PMID- 11421919 TI - Identification of four novel potato (Solanum tuberosum) allergens belonging to the family of soybean trypsin inhibitors. AB - BACKGROUND: We have previously identified patatin (Sol t 1) of potato tubers as a major food allergen among atopic children. In addition to Sol t 1, concomitant IgE binding to other, then unidentified, potato proteins was observed. METHODS: Purification and identification of the putative allergens were done by both standard and advanced methods of protein chemistry. The patient series comprised 39 children with positive skin prick test (SPT) to raw potato. Immunoblotting and ELISA were used to examine IgE-binding ability and skin prick testing to assess in vivo reactivity of the purified potato proteins. RESULTS: Four IgE-binding potato proteins with molecular masses ranging from 16 to 20 kDa were purified and identified as cathepsin D-, cysteine-, and aspartic protease inhibitors belonging to the family of soybean trypsin inhibitors (Kunitz type). The proteins were designated Sol t 2, Sol t 3.0101, Sol t 3.0102, and Sol t 4. In ELISA, 51% of the sera of the 39 atopic children showed specific IgE to Sol t 2, 43% to Sol t 3.0101, 58% to Sol t 3.0102, and 67% to Sol t 4, respectively. All these four allergens were able to produce positive wheal-and-flare responses in SPT. CONCLUSION: In addition to Sol t 1, potato tubers contain several proteins belonging to the family of soybean trypsin inhibitors against which atopic children with positive SPT responses to raw potato have in vitro and in vivo reactive IgE antibodies. PMID- 11421920 TI - Nasal and exhaled nitric oxide in response to occupational latex exposure. AB - BACKGROUND: Latex sensitivity is an increasing problem among health-care workers. Allergic responses are associated with changes in nitric oxide (NO) generation and the changes secondary to latex have not been described. METHODS: A total of 22 subjects comprising equal numbers of control volunteers and subjects with self reported latex sensitivity were recruited to undergo latex skin prick testing. Symptom scores, exhaled nitric oxide (eNO), and nasal nitric oxide (nNO) were studied 1) before and after a controlled latex challenge (n=16), and 2) at the beginning and end of the working week, during exposure to latex (n=18). RESULTS: Latex challenge caused a significant fall in nNO levels in latex-sensitive subjects, compared to normal control subjects (P=0.04). eNO levels also decreased in the latex-sensitive subjects after latex challenge, but to a lesser degree. There were no significant differences between the beginning and end of the working week in terms of eNO or nNO in either group, although symptom scores showed a nonsignificant increase in latex-sensitive subjects. CONCLUSION: Fall in nasal NO after latex challenge is associated with reported symptomatic latex sensitivity, and this corresponds to latex skin prick test positivity. Neither nNO nor eNO showed a clear relationship to routine workplace exposure. PMID- 11421921 TI - Polyunsaturated fatty acids in maternal diet, breast milk, and serum lipid fatty acids of infants in relation to atopy. AB - BACKGROUND: The increased consumption of n-6 polyunsaturated fatty acids (PUFA) has been shown to coincide with the increased prevalence of atopic diseases. We aimed to investigate whether maternal diet and atopic status influence the PUFA composition of breast milk and the serum lipid fatty acids of infants. METHODS: Maternal diet was assessed by a food questionnaire. The PUFA composition of breast milk obtained at 3 months from 20 allergic and 20 healthy mothers and of their infants' (10 atopic and 10 nonatopic/group of mothers) serum lipids was analyzed. RESULTS: Although no differences in maternal PUFA intake were observed, the breast milk of allergic mothers contained less gamma-linolenic acid (18:3 n 6) than that of healthy mothers. Similarly, atopic infants had less gamma linolenic acid in phospholipids than healthy infants, although n-6 PUFA were elevated in other serum lipid fractions in atopic infants. The serum lipid fatty acids in atopic infants did not correlate with those in maternal breast milk. CONCLUSION: Our results suggest that dietary n-6 PUFA are not as readily transferred into breast milk or incorporated into serum phospholipids, but may be utilized for other purposes, such as eicosanoid precursors, in allergic/atopic individuals. Subsequently, high dietary proportions of n-6 PUFA, or reduced proportions of regulatory PUFA, such as gamma-linolenic acid and n-3 PUFA, may be a risk factor for the development of atopic disease. PMID- 11421922 TI - The mechanisms of serum-treated zymosan (STZ)-induced oxidative metabolism by human eosinophils and the effects of IL-5 priming. AB - BACKGROUND: The aim of this work was to study the mechanisms of action of IL-5 on the subsequent stimulation of the oxidative metabolism of blood eosinophils by serum-treated zymosan (STZ), in terms of signal transduction characteristics, and by comparing the response of cells from healthy and allergic subjects during environmental exposure to birch pollen. METHODS: Eosinophils from healthy controls and allergic patients were purified to over 95% by Percoll gradients and the MACS system. Oxidative metabolism was measured by a lucigenin-enhanced chemiluminescence (CL) assay. Eosinophils were primed with IL-5 and subsequently stimulated with STZ. The signal transduction mechanisms of IL-5 priming were studied with the MEK inhibitor PD 98059,the PkC inhibitors staurosporine and Ro 318220, and the PI3 kinase inhibitor wortmannin. RESULTS: IL-5 increased the maximum radical production (P=0.0079) and reduced the t(1/2) rise (0.000018) of the CL reactions. The t(1/2) rise was PkC dependent and MEK independent, while the maximum radical production was PkC, MEK, and PI3 kinase dependent. During the pollen season, IL-5 reduced the total STZ-induced CL response in the patients' cells (P=0.016), but not in the control cells, whereas it primed the response to STZ of both cell populations in terms of the t(1/2) rise (P=0.012 and 0.00066, respectively). CONCLUSION: STZ-induced oxidative metabolism consists of different stages. The initial stage (t(1/2) rises of the curves) is PkC dependent and MEK independent, while the end stage (maximum radical production) is PkC, MEK, and PI3 kinase dependent. IL-5 shortened the initial stage, and increased the end stage. During allergen exposure, however, the end stage was reduced by IL-5. This could be due to increased amounts of hypodense eosinophils and/or some abnormality in cell responses. PMID- 11421923 TI - Influence of early and current environmental exposure factors on sensitization and outcome of asthma in pre-school children. AB - BACKGROUND: Exposure to furred pets in early life has been considered to increase the risk of allergic sensitization and consequent development of asthma later in children. However, recently, it has been suggested that early exposure to pets prevents sensitization. The aim of this study was to evaluate the importance of early exposure to pets and other environmental risk factors in asthmatic children. METHODS: This is a follow-up study after 2 years of a previously investigated group of 193 asthmatic children, aged 1-4 years. The study was completed by 181 children, who were clinically examined; serum IgE antibodies were also measured and a questionnaire was answered. RESULTS: Children with reported exposure to cats during the first 2 years of life were more likely to have developed sensitization to cat by 4 years of age than unexposed children. High levels of cat allergen (Fel d 1> or =8 microg/g dust) were associated with an increased risk of sensitization to cat and, in combination with tobacco smoke, also with the development of more severe asthma. CONCLUSION: In young asthmatic children, early exposure to cat and tobacco smoke increased the risk of allergic sensitization and further development of more severe asthma later in childhood. PMID- 11421924 TI - Mizolastine in the treatment of seasonal allergic rhinoconjunctivitis: a European clinical experience with 5408 patients managed in daily practice (PANEOS SAR Study). AB - BACKGROUND: Mizolastine, a potent H1 antihistamine with additional antiallergic properties, is marketed for the treatment of allergic rhinoconjunctivitis and urticaria. The objective was to investigate the safety and effectiveness of mizolastine under conditions of daily practice in patients with seasonal allergic rhinoconjunctivitis (SAR). METHODS: In an open multicenter study, mizolastine 10 mg daily was administered for 14 days during the pollen season. Nasal and ocular symptoms, time to onset of symptom relief, and effect of the drug on diurnal alertness were evaluated. Safety was evaluated on the basis of self-reported adverse events (AE). RESULTS: A total of 5408 patients (36+/-14 years of age, females=57%) with a history of SAR for 8+/-9 years were treated for a mean of 17.1+/-5.0 days. SAR symptoms improved in 93% and decreased by at least 50% in 86% of patients; 78% reported improvement after the first drug intake and 51% from the first hour. Sixty-nine percent considered mizolastine more effective than other antihistamines taken previously. The incidence of AE was low (3.8%). CONCLUSION: The high responder rate, the rapid onset of action, and the low incidence of AE observed in this large multicenter study confirm the previously reported beneficial efficacy and safety of mizolastine in the management of SAR. PMID- 11421925 TI - Allergenic cross-reactivity between the nematode Anisakis simplex and the dust mites Acarus siro, Lepidoglyphus destructor, Tyrophagus putrescentiae, and Dermatophagoides pteronyssinus. AB - BACKGROUND: The nematode Anisakis simplex is a common parasite on fish and other seafood. It is considered to be a food allergen and to induce IgE-mediated reactions. Allergenic cross-reactivity between A. simplex and other nematodes has been reported, as has cross-reactivity with arthropods: red mosquito larvae and German cockroach. We have here studied the allergenic relationship between A. simplex and four different dust-mite species. METHODS: Serum samples collected from 69 farmers allergic to dust mites were analyzed for IgE to A. simplex by CAP FEIA. Allergenic cross-reactivity between A. simplex and dust mites was studied in two of the sera by CAP FEIA and immunoblotting inhibition. RESULTS: We found that 14/69 farmers had detectable levels of IgE antibodies to A. simplex. The IgE response in CAP FEIA to A. simplex was inhibited to various degrees in the two studied sera by extracts of the dust mites Acarus siro, Lepidoglyphus destructor, Tyrophagus putrescentiae, and Dermatophagoides pteronyssinus. In the reverse inhibition experiment, extract of A. simplex inhibited the response in both sera to A. siro and T. putrescentiae, but not to L. destructor. The IgE binding to D. pteronyssinus was inhibited in one of the two sera. In blotting inhibition experiments, the IgE binding to several allergens in A. simplex was inhibited by each of the four mite extracts, especially by A. siro and T. putrescentiae, which completely inhibited the IgE binding to several allergens. CONCLUSION: The results show allergenic cross-reactivity between several allergens in A. simplex and four dust-mite species. The clinical significance of this cross-reactivity remains to be evaluated. PMID- 11421926 TI - Prevalence of and risk factors for IgE sensitization to Anisakis simplex in a Spanish population. AB - BACKGROUND: The number of allergic reactions to A. simplex reported in Spain has increased dramatically in the last decade. Nevertheless, there have been no studies of the prevalence of and possible risk factors for IgE sensitization to this parasite, possibly because suitably specific diagnostic methods have only recently become available. The objective was to investigate the prevalence of and risk factors for IgE sensitization to A. simplex in Galicia, a region of northwestern Spain with a population of about 3 million and high average fish consumption (78.5 g/person per day). METHODS: The study was performed with a random sample of 2801 healthy blood donors distributed in 53 geographic areas, proportional to the density of donors. IgE sensitization to A. simplex was tested by a capture ELISA method that has proved to be the most specific method currently available. RESULTS: The results showed a total of only 12 positive subjects, of whom five also showed IgG1 sensitization. All positive subjects and 101 randomly selected seronegative subjects were then included in a case-control study of risk factors for sensitization to A. simplex, based on a telephone interview about fish consumption (especially raw and undercooked fish). All seropositive subjects (but only 25% of seronegative subjects) reported consumption of undercooked fish or homemade raw-fish products. CONCLUSIONS: Our results strongly suggest that sensitization to A. simplex is caused only by live larvae, and not by allergens contained in fish tissues, and that ingestion of homemade boquerones (anchovies [Engraulis encrasicholus] in vinegar), and to a much lesser extent of undercooked fish, are the main risk factors for IgE sensitization to Anisakis in this region. PMID- 11421927 TI - Expression of adhesion molecules on cord-blood-derived, cultured human mast cells and effect of dexamethasone on intercellular adhesion molecule-1 expression on the mast cells treated by phorbol myristate acetate. AB - BACKGROUND: Increase in mast-cell number at sites of allergic inflammation has been observed, and glucocorticoids applied to the sites have been shown to result in a significant reduction in mast cells. However, the expression of adhesion molecules on cultured human mast cells and their regulation by glucocorticoids is poorly understood. METHODS: Cultured human mast cells were raised from human umbilical cord-blood cells, and the expression of adhesion molecules on the mast cells was analyzed by flow cytometry. The cells were also incubated with 10 ng/ml phorbol myristate acetate (PMA) for the indicated time, and the effect of dexamethasone on adhesion molecule expression on PMA-treated, cultured human mast cells was examined. RESULTS: Cord-blood-derived, cultured human mast cells constitutively expressed intercellular adhesion molecule-1 (ICAM-1), very late antigen-4 (VLA-4), and macrophage-1 antigen (Mac-1). Weak expression of lymphocyte function-associated antigen-1 (LFA-1) was observed on the cells, whereas they failed to express vascular cell adhesion molecule-1 (VCAM-1). Kinetic studies showed that after a transient downregulation reaching a minimum at 8 h, the expression of ICAM-1 was markedly upregulated on PMA-treated mast cells after a 24-h incubation. In contrast, the expression of VLA-4 and Mac-1 was decreased after the incubation with PMA for 24 h. The PMA-induced upregulation of ICAM-1 was inhibited by dexamethasone in a concentration-dependent manner. CONCLUSION: Our results indicate that cord-blood-derived, cultured human mast cells constitutively express integrins and ICAM-1, but not VCAM-1, and demonstrate for the first time that dexamethasone inhibits the upregulation of ICAM-1 on PMA-treated, cultured human mast cells. PMID- 11421928 TI - Allergy to Diplotaxis erucoides pollen: occupational sensitization and cross reactivity with other common pollens. AB - BACKGROUND: Diplotaxis erucoides is a common weed of the Brassicaceae family widespread in southern and central Europe. METHODS: A total of 410 consecutive patients referred for allergy study of rhinoconjunctivitis and/or asthma were skin tested with D. erucoides pollen, 14 proving positive. A purified D. erucoides pollen extract was prepared to perform quantitative skin tests, provocation tests, immunoblotting, and EIA inhibition in the 14 sensitized patients. RESULTS: Three patients, directly involved in viniculture, had rhinoconjunctivitis related to D. erucoides pollen. No D. erucoides-related symptoms were observed in most patients, who were also sensitized to Artemisia pollen. RAST was positive in 12/14 patients and nasal provocation tests in 9/12. The molecular masses of the most prevalent IgE-binding proteins ranged from 26 to 27.5 and from 31 to 34 kDa. D. erucoides pollen inhibited the IgE-binding of other sensitizing pollens in the three viniculture workers, whereas both Artemisia and D. erucoides pollen produced similar heterologous inhibition in the pooled serum of the remaining, nonclinically affected, D. erucoides-sensitized patients. CONCLUSION: D. erucoides pollen may be an important prevalent aeroallergen, particularly in rural areas. It may act as an occupational allergen in vineyard workers, in whom it seems to be the primary sensitizing agent, playing a secondary cross-reactive role in other sensitized patients. PMID- 11421929 TI - Sensitization to the fungus Hemileia vastatrix (coffee leaf rust). AB - BACKGROUND: The aim of the present study was to determine the existence of sensitization to the fungus Hemileia vastatrix (coffee leaf rust) in a sample of the Brazilian population. This fungus attacks coffee plantations, and in some regions of Brazil the concentration of its spores in the air can be very high. METHODS: A total of 378 individuals underwent skin tests (prick and intradermal tests) with H. vastatrix extract. The subjects were divided into four groups according to the occurrence of atopy and the region where they lived (coffee growing or non-coffee-growing regions), and another group (V) consisted of 50 rural workers employed on coffee plantations. The presence of specific IgE against the fungus H. vastatrix in sensitized individuals was demonstrated in vitro by immunoblotting. RESULTS: There was no statistical difference in comparing the results with intradermal or prick tests. The incidence of positive tests was significantly higher among atopic individuals residing in coffee growing regions (14.7%). Among rural workers, 10% showed positive tests. Immunoblotting revealed specific IgE against 20 protein bands of H. vastatrix. CONCLUSION: The fungus H. vastatrix may be an important allergen in coffee producing countries. The sensitization is more frequent among atopic individuals residing in coffee-growing regions. PMID- 11421930 TI - Cysteinyl-leukotriene release test (CAST) in the diagnosis of immediate drug reactions. AB - BACKGROUND: The diagnosis of allergic reactions to drugs is difficult. Most skin tests are not standardized, and in vitro tests are needed to avoid provocation tests. Cross-linking of IgE on basophils is known to cause the release of both cysteinyl leukotriene (Cys-LT) and histamine. We aimed to evaluate the diagnostic utility (sensitivity, specificity, and efficiency) of measurement of sulfidoleukotrienes in drug allergy. METHODS: We performed a prospective study in 55 patients with proven immediate adverse reactions to drugs (30 to beta-lactams, six to acetaminophen, and 19 to aspirin) and 64 drug-exposed nonallergic controls. Positive diagnosis was established by history, skin tests, and, if needed, oral provocation tests. Cys-LT release was determined after drug-allergen stimulation by the cellular antigen stimulation test (CAST(R)) technique. Histamine release was also assessed on the same samples by enzyme immunoassay. Spontaneous and anti-FcepsilonRIalpha-induced mediator release was also studied in all subjects. Sensitivity, specificity, and efficiency were calculated. RESULTS: Net Cys-LT release was over the maximal threshold given by the manufacturer in 19/55 patients and in 9/64 controls. Net histamine release was over 5% of total histamine content in 28/55 patients and 34/64 controls. The efficiency of both tests was low. CONCLUSION: Thus, in most cases, the in vitro Cys-LT test has little or no diagnostic utility and is not superior to histamine release. PMID- 11421931 TI - Increased serum nitrate levels in infants with atopic dermatitis. AB - BACKGROUND: The pathogenesis of atopic dermatitis (AD) is still unknown. A recent study has shown that inducible nitric oxide synthase (iNOS) is expressed in the atopic skin lesion, suggesting the involvement of nitric oxide in the skin inflammation of AD. The purpose of the study was to examine serum nitrate (NO3) levels in relation to the disease severity in children with AD. METHODS: Serum nitrate levels were assessed in relation to the skin scores in 88 patients with atopic dermatitis (AD) (aged 0.4-8 years: mean+/-SD, 2.2+/-1.9, 41 boys and 47 girls) and 12 nonatopic children (aged 0.8-4 years: mean+/-SD, 1.8+/-0.9, seven boys and five girls). RESULTS: Serum nitrate levels of patients with AD were significantly increased as compared to nonatopic controls and were also correlated with the disease severity. The skin scores were significantly correlated with serum nitrate levels as well as peripheral eosinophil counts. CONCLUSION: Our results indicate that nitric oxide may be involved in the pathogenesis of vasodilation and erythema in AD skin. PMID- 11421932 TI - A baker's occupational allergy to flour moth (Ephestia kuehniella). AB - BACKGROUND: Allergy to insects is common. However, few reports cover occupational sensitization to flour moth (Ephestia [syn. Anagasta] kuehniella). We describe a baker who suffered from IgE-mediated occupational respiratory allergy to flour moth. METHODS: The skin prick test (SPT) and serum IgE tests were used to evaluate the patient's sensitivity to flour moth. Allergen cross-reactivity with mites was evaluated in IgE-inhibition studies. Clinical sensitivity was evaluated by nasal challenge test. Pulmonary function tests were repeatedly monitored. RESULTS: SPT with flour moth gave a 6-mm wheal, and an elevated level of flour moth-specific IgE was measured in the patient's serum (1.9 PRU/ml, RAST class 2). Immunoblotting with the patient's serum revealed at least seven heavy IgE-binding bands with molecular masses of 22, 35, 43, 53, 65, 77, and >86 kDa in the extract of flour moth. Allergen cross-reactivity with mites was demonstrated in inhibition studies. Immediate-type allergy to flour moth was confirmed by nasal challenge. Increased daily variability of PEF values was observed during workplace exposure. CONCLUSION: A baker's occupational respiratory allergy to flour moth was confirmed. PMID- 11421933 TI - Allergy to tetanus toxoid vaccine. PMID- 11421934 TI - Introduction: leukotriene receptor antagonists--exciting new approach to the management of childhood asthma. PMID- 11421935 TI - Pathophysiology of the cysteinyl leukotrienes and effects of leukotriene receptor antagonists in asthma. AB - Cysteinyl leukotrienes, synthesized de novo from cell membrane phospholipids, are proinflammatory mediators that play an important role in the pathophysiology of asthma. These mediators are among the most potent of bronchoconstrictors and cause vasodilation, increased microvascular permeability, exudation of macromolecules and edema. The cysteinyl leukotrienes also have potent chemoattractant properties for eosinophils, causing an influx of eosinophils into the airway mucosa, which further fuels the inflammatory process. In addition, the cysteinyl leukotrienes are potent secretagogues and reduce ciliary motility, which may hinder mucociliary clearance. Asthmatic patients demonstrate increased production of cysteinyl leukotrienes during naturally occurring asthma and acute asthma attacks as well as after allergen and exercise challenge. The leukotriene receptor antagonists montelukast, zafirlukast and pranlukast inhibit bronchoconstriction in asthmatic patients undergoing allergen, exercise, cold air or aspirin challenge. They attenuate the hallmarks of asthmatic inflammation, including eosinophilia in the airway mucosa and peripheral blood. Moreover, exhaled nitric oxide concentrations, another correlate of airway inflammation, are decreased during montelukast treatment in children. Cysteinyl leukotriene synthesis is not blocked by corticosteroid therapy. This important observation suggests that the leukotriene receptor antagonists represent a novel therapeutic approach, one that may provide benefits that are additive with corticosteroid therapy. This supposition is supported by clinical observations that treatment with leukotriene receptor antagonists significantly improve asthma control when added to inhaled corticosteroid therapy. Moreover, the bronchodilator properties of the leukotriene receptor antagonists are additive with those of beta agonists. These data provide strong support for the use of leukotriene receptor antagonists for treating asthma. PMID- 11421936 TI - Choices of therapy for exercise-induced asthma in children. AB - Cough and wheezing interferes with sport and other forms of physical activity in half of asthmatic children. Airway obstruction can be induced by a standard exercise test in over 70% of children with asthma. A beta-adrenergic agonist or cromone taken by inhalation beforehand will usually inhibit bronchoconstriction provoked by a free running exercise test. The duration of protective effect with salbutamol, terbutaline and cromones is less than 4 h. The long acting beta adrenergic agonists formoterol and salmeterol give protection against exercise induced airway obstruction for up to 12 h, which implies that treatment given in the morning will offer protection from the effects of physical activity throughout the day. However, the duration of protective effect after a morning dose declines if these compounds are given regularly. Leukotriene receptor antagonists (LRAs) also provide good protection against exercise-induced asthma. Regular administration of LRAs is not associated with tolerance and loss of protective effect. The oral route makes for unobtrusive administration and this may help adherence to prescribed regimens. PMID- 11421937 TI - Issues and challenges in pediatric asthma. PMID- 11421938 TI - The role of leukotriene receptor antagonists in the treatment of chronic asthma in childhood. AB - A considerable increase in the prevalence of childhood asthma over the last few decades has been mirrored by a dramatic increase in usage of anti-asthma drugs; however, there has been no reduction in the numbers of patients dying of asthma. Concern has been expressed about the development of tolerance with continuous use of inhaled beta-agonist bronchodilators and about the potential adverse systemic effects of high-dose inhaled corticosteroids in children. Moreover, patient compliance with inhaled therapy tends to be poor. The leukotriene receptor antagonists, including montelukast, pranlukast and zafirlukast, are orally administered agents with proven benefits in asthma. In a large, placebo controlled pediatric trial, montelukast significantly (P < 0.02) reduced requirements for rescue beta-agonist bronchodilators, improved quality of life, reduced the circulating level of blood eosinophils and produced improvements in lung function. In adult studies, montelukast reduced sputum eosinophils and attenuated early and late phase allergen-induced reactions. Montelukast has also demonstrated protective effects against exercise-induced bronchospasm in both adults and children, and this protection was maintained during the trough period at the end of the once-daily administration interval (namely, 20-24 h post-dose). Several studies have demonstrated that the formation of cysteinyl leukotrienes in the airways of asthmatic patients is not suppressed by corticosteroids; thus, it is not surprising that montelukast demonstrates complementary effects when given with inhaled corticosteroids. Currently, the most compelling evidence from published trials suggests that leukotriene receptor antagonists can be used as add-on therapy to inhaled corticosteroids to allow tapering of corticosteroid dose and reduction in beta-agonist use. Recent clinical trial results suggest there may also be a role for these agents as first-line therapy in children with mild asthma. PMID- 11421939 TI - Breed demarcation and potential for breed allocation of horses assessed by microsatellite markers. AB - Population demarcation of eight horse breeds was investigated using genotype information of 306 horses from 26 microsatellite loci. The breeds include the indigenous Norwegian breeds Fjord Horse, Nordland/Lyngen Horse, Dole Horse and Coldblooded Trotter together with Icelandic Horse, Shetland Pony, Standardbred and Thoroughbred. Both phylogenetic analysis and a maximum likelihood method were applied to examine the potential for breed allocation of individual animals. The phylogenetic analysis utilizing simple allele sharing statistics revealed clear demarcation among the breeds; 95% of the individuals clustered together with animals of the same breed in the phylogenetic tree. Even breeds with a short history of divergence like Dole Horse and Coldblooded Trotter formed distinct clusters. Implementing the maximum likelihood method allocated 96% of the individuals to their source population, applying an assignment stringency of a log of the odds ratio larger than 2. Lower allocation stringency assigned nearly all the horses. Only three individuals were wrongly allocated a breed by both methods. In conclusion, the study demonstrates clear distinction among horse breeds, and by combining the two assignment methods breed allocation could be determined for more than 99% of the individuals. PMID- 11421940 TI - Mapping of expressed sequence tags from a porcine early embryonic cDNA library. AB - The goal of this study was to identify and map genes expressed during the elongation phase of embryogenesis in swine. Expressed sequence tags were analysed from a previously described porcine cDNA library prepared from elongating swine embryos. Average insert length of randomly selected clones was approximately 600 bp, with a range from < 100 to > 2500 bp. Single-pass, coding strand sequences from 1132 independent clones were compared with the GenBank non-redundant (nr) database via BLASTN analysis to identify potential porcine homologous of known genes. Among these sequences, 781 (69%) showed significant (score > 300) homology to non- mitochondrial sequences previously deposited in GenBank. Sequences matching interleucin 1 beta and thymosin beta 10 were most frequently observed (24 and 18 clones, respectively), in addition to matches with 310 other distinct genes. No significant match in the GenBank nr database was obtained for 303 sequences. Analysis demonstrated that 151 (50%) had open reading frames (ORF) extending at least 50 codons from the first base of the clone insert. Genetic markers were developed and used to map a subset of 17 genes, selected on the basis of function or of the ability to design primers that successfully amplified porcine genomic DNA, to 10 different porcine chromosomes, providing a set of mapped markers corresponding to genes expressed during conceptus elongation. PMID- 11421941 TI - cDNA cloning and genetic polymorphism of the swine major histocompatibility complex (SLA) class II DMA gene. AB - cDNA clones corresponding to the swine histocompatibility complex (SLA: swine leucocyte antigen)-DM alpha chain were isolated using the polymerase chain reaction (PCR) products from the third exon in the human HLA-DMA gene as a probe. Amino acid comparative analysis revealed that these clones were more closely related to the bovine and human DMA genes than to the other swine class II genes alpha chain genes, DRA, DQA and DOA. These results suggest that the SLA-DMA gene is expressed and may function, like HLA-DM, as an important modulator in class II restricted antigen processing in swine. Furthermore, based on the sequences and PCR-restriction fragment length polymorphism (PCR-RFLP) patterns in the SLA-DMA gene, no allelic variation was recognized in the second exon, but five allelic variations were recognized in the third exon in five different breeds of swine. These DMA alleles were defined by variation at four nucleotide positions. Two of these alleles resulted in an amino acid substitution. These results suggest that SLA-DMA has little polymorphism as observed in HLA-DMA and mouse H2-Ma. PMID- 11421942 TI - Polymorphism identification within 50 equine gene-specific sequence tagged sites. AB - The continued discovery of polymorphisms in the equine genome will be important for future studies using genomic screens and fine mapping for the identification of disease genes. Segments of 50 equine genes were examined for variability in 10 different horse breeds using a pool-and-sequence method. We identified 11 single nucleotide polymorphisms (SNPs) in 9380 bp of sequenced exon, and 25 SNPs, six microsatellites, and one insertion/deletion in 16961 bp of sequenced intron. Of all genes studied 52% contained at least one polymorphism, and polymorphisms were found at an overall rate of 1/613 bp. Several of the putative SNPs were tested and verified by restriction enzyme analysis using natural restriction sites or ones created by primer mutagenesis. The lowest allele frequency for a SNP detected in pooled samples was 10%. Three of the SNPs verified in the diverse horse pool were further tested in six breed-specific horse pools and were found to be reasonably variable within breeds. The pool-and-sequence method allows identification of polymorphisms in horse populations and will be a valuable tool for future disease gene and comparative mapping in horses. PMID- 11421943 TI - Differential introgression of uniparentally inherited markers in bison populations with hybrid ancestries. AB - Historical hybridization between Bison bison (bison) and Bos taurus (cattle) has been well documented and resulted in cattle mitochondrial DNA (mtDNA) introgression, previously identified in six different bison populations. In order to examine Y chromosome introgression, a microsatellite marker (BYM-1) with non overlapping allele size distributions in bison and cattle was isolated from a bacterial artificial chromosome (BAC) clone, and was physically assigned to the Y chromosome by fluorescence in situ hybridization. BYM-1 genotypes for a sample of 143 male bison from 10 populations, including all six populations where cattle mtDNA haplotypes were previously identified, indicated that cattle Y chromosome introgression had not occurred in these bison populations. The differential permeability of uniparentally inherited markers to introgression is consistent with observations of sterility among first generation hybrid males and a sexual asymmetry in the direction of hybridization favouring matings between male bison and female cattle. PMID- 11421944 TI - Comprehensive linkage map of bovine chromosome 11. AB - The results of genotypic data contributed to the International Society of Animal Genetics (ISAG) Bovine Chromosome 11 (BTA11) Workshop are presented. Six laboratories contributed a total of 26 199 informative meioses from 80 loci. Thirty-six loci were typed by at least two independent laboratories and were used to construct a consensus linkage map of the chromosome. The remaining loci were subsequently incorporated into a comprehensive map. The sex-averaged consensus map covered 128.9 cM. The female consensus map was 101.2 cM, while the male consensus map was 129.8 cM. The comprehensive sex-averaged map was 134.2 cM and the average genetic distance between loci was 1.72 cM. PMID- 11421945 TI - Comprehensive linkage map of bovine chromosome 27. AB - The results of genotypic data contributed to the International Society for Animal Genetics (ISAG) Bovine Chromosome 27 Workshop are presented. Eight laboratories contributed 23 261 informative meioses from 44 loci. Eighteen loci were typed by at least two laboratories and were used to construct a consensus linkage map. Twenty-one loci were subsequently incorporated into a comprehensive map. The sex averaged consensus map covered 66.9 cM. The sex-averaged comprehensive map was 75.5 cM, while the female and male maps were 73.1 and 63.7 cM, respectively. Five loci were excluded from the analysis because of ambiguous position in the linkage group and a low LOD score (less than 2.0). Average distance between loci in the comprehensive map was 1.98 cM. PMID- 11421946 TI - Linked markers exclude KIT as the gene responsible for appaloosa coat colour spotting patterns in horses. AB - The appaloosa coat colour pattern of the horse is similar to that caused by the rump-white (Rw) gene in the mouse. In the mouse Rw colour pattern is the result of an inversion involving the proto-oncogene c-kit (KIT). Therefore, we investigated KIT as a candidate gene that encodes the appaloosa coat colour gene (Lp) in horses. KIT plays a critical role in haematopoiesis, gametogenesis, and melanogenesis and encodes a transmembrane tyrosine kinase receptor that belongs to the PDGF/CSF-1/c-KIT receptor subfamily. Half-sib families segregating for Lp were uninformative for a reported polymorphism in KIT. However, KIT is located on horse chromosome 3 close to albumin (ALB), serum carboxylesterase (ES), vitamin D binding protein (GC) and microsatellite markers ASB23, LEX007, LEX57, and UCDEQ437. Indeed, KIT and ASB23 were localized to ECA3q21-22.1 and 3q22.1-22.3, respectively, by fluorescent in situ hybridization. Family studies were conducted to investigate linkage of Lp to these markers using eight half-sib families in which Appaloosa stallions were mated to solid coloured mares. Linkage of Lp to the chromosome region containing ES, ALB, GC, ASB23, UCDEQ437, LEX57, and LEX007 was investigated by a multipoint linkage analysis using the computer program GENEHUNTER. LOD scores over the interval under investigation ranged from -4.28 to -12.48, with a score of -12.48 at the location for ASB23. Therefore, it was concluded that appaloosa (Lp) is not linked to any of the tested markers on ECA3, and thus Lp is unlikely to be the product of KIT. PMID- 11421947 TI - Physical mapping of CSF2RA, ANT3 and STS on the pseudoautosomal region of bovine chromosome X. AB - The pseudoautosomal region (PAR) of bovine chromosome X (BTA X) has a particularly low representation of genes and markers, making comparative gene mapping in this region difficult. We describe the localization of three genes, colony-stimulating factor 2 receptor alpha (CSF2RA), ADP/ATP translocase 3 (ANT3) and steroid sulphatase (STS) on PAR of BTA X using a 5000 rad whole-genome radiation hybrid panel. The relationship of these genes to a number of previously mapped simple sequence repeat (microsatellite) markers is determined by physical and radiation hybrid mapping methods. The resulting radiation hybrid map resolves a discrepancy between the two major bovine linkage maps in the PAR of BTA X. PMID- 11421948 TI - Identification of two polymorphic microsatellite repeats on a bovine TNFA containing BAC clone. PMID- 11421949 TI - Linkage mapping of four genes (OTC, SERPINA7, SLC25A5 and FMR1) on porcine chromosome X. PMID- 11421950 TI - The porcine sarcolipin (SLN) gene: identification of an SNP and linkage mapping to chromosome 9. PMID- 11421951 TI - Linkage and radiation hybrid mapping of the porcine gene for subunit C of succinate dehydrogenase complex (SDHC ) to chromosome 4. PMID- 11421952 TI - Consensus and comprehensive linkage maps of bovine chromosome 17. PMID- 11421953 TI - Consensus and comprehensive linkage maps of bovine chromosome 25. PMID- 11421954 TI - Consensus and comprehensive linkage maps of the bovine sex chromosomes. PMID- 11421955 TI - Characterization of equine microsatellite loci, TKY102-TKY112. PMID- 11421956 TI - COM184: a polymorphic microsatellite marker for CKB on chicken chromosome 5. PMID- 11421957 TI - Linkage mapping of the tyrosinase gene to bovine chromosome 29. PMID- 11421958 TI - Assignment of the SERPINA7 gene to porcine chromosome band Xq22 by FISH. PMID- 11421959 TI - Aboriginal health. PMID- 11421960 TI - Development of oral health training for rural and remote aboriginal health workers. AB - Research data exists that highlight the discrepancy between the medical/dental status experienced by Aboriginal people compared with that of their non Aboriginal counterparts. This, coupled with a health system that Aboriginal people often find alienating and difficult to access, further exacerbates the many health problems they face. Poor oral health and hygiene is an issue often overlooked that can significantly impact on a person's quality of life. In areas where Aboriginal people find access to health services difficult, the implementation of culturally acceptable forms of primary health care confers significant benefits. The Aboriginal community has seen that the employment and training of Aboriginal health workers (AHW), particularly in rural and remote regions, is significantly beneficial in improving general health. In the present study, an oral health training program was developed and trialled. This training program was tailored to the needs of rural and remote AHWs. The primary objective was to institute a culturally appropriate basic preventative oral health delivery program at a community level. It is envisaged that through this dental training program, AHWs will be encouraged to implement long-term preventive measures at a local level to improve community dental health. They will also be encouraged to pursue other oral health-care delivery programs. Additionally, it is considered that this project will serve to strengthen a trust-based relationship between Aboriginal people and the health-care profession. PMID- 11421961 TI - Aboriginal health: a sick minority skews statistics. AB - Overall morbidity and mortality rates are higher among Aboriginal people than in the general Australian population. It is unclear, however, whether this is because of general poor health or a minority with very high levels of disease within a reasonably healthy Aboriginal community. At Cherbourg Community, almost all health care is given by the hospital. All visits to the casualty/outpatient departments at the hospital have been recorded since 1993. These data have been analysed to find the patterns of visiting by person, by diagnosis and by year. Over the 5 years with complete data, most people made less than five visits per year (low visit group) to the hospital ambulatory clinic, but at the other extreme, approximately 14% of people made 50% of the visits (high visit group). People who made many visits in 1 year made approximately three times as many visits as the low visit group in other years. When the high and low visit groups were compared, the high visit group was more likely to be female and to come for problems related to infection, but were less likely to come with injuries or accidents. The age distribution was similar in both groups. If the number of visits to a health service is an indicator of health, then most Aboriginal people at Cherbourg have reasonable health, but a sizeable minority have health problems that extend over several years. This group skews the overall statistics of Aboriginal health and should be targeted for extra attention. PMID- 11421962 TI - An introduction to indigenous health and culture: the first tier of the Three Tiered Plan. AB - The objective of the present study was to prepare new doctors with an awareness of cultural and health issues to facilitate positive experiences with indigenous patients. The study incorporated the 1998 intern orientation programs in Queensland public hospitals. The study method included tier one of the Three Tiered Plan, which was implemented and audited. Indigenous liaison officers, directors of clinical training and medical education officers were surveyed prior to this implementation to determine whether any or similar initiatives had been carried out in previous years and/or were planned. Post-implementation feedback from interns was obtained by using questionnaires. Follow-up telephone interviews with the directors of clinical training, medical education officers and indigenous hospital liaison officers detailed the format and content of tier one at each hospital. The results indicate that this active intervention improved the implementation rate of tier one from nine of 19 (47%) Queensland public hospitals in 1997 to 17 (90%) in 1998. The 14 indigenous hospital liaison officers (100%) involved in the intervention perceived it as beneficial. Forty-three (67%) of interns who responded to the survey indicated they had encountered an indigenous patient within the last 2-4 months. The level of knowledge of indigenous health and culture self-reported by interns was between the categories 'enough to get by' and 'inadequate'. In conclusion, it appears that tier one has been successful and is to be a formal component of intern orientations in Queensland public hospitals. Further initiatives in indigenous health and culture targeting medical staff (i.e. tier two and tier three), are needed. PMID- 11421963 TI - Diabetic foot care: developing culturally appropriate educational tools for Aboriginal and Torres Strait Islander peoples in the Northern Territory, Australia. AB - Evidence shows that Aboriginal and Torres Strait Islander people have the highest national percentage of morbidity in relation to diabetes. Aboriginal and Torres Strait Islander people also suffer the greatest risk of amputation as a complication of diabetes. This participatory action research project sought to discover the opinions of a range of people, including registered nurses, general practitioners, Aboriginal health workers, cross-cultural liaison officers and Aboriginal and Torres Strait Islander people with diabetes. Focus groups provided valuable information regarding relevant issues of foot care education in the Northern Territory. The emergent themes included communication issues, educational resources, changing behaviour and other practical resources required for health education. The themes provided evidence of the inherent issues of foot care for Aboriginal and Torres Strait Islander people and guidance for the development of a visual educational tool. The results have lead to the development of a foot care educational tool that will be used by health-care professionals and clients in urban, community, rural and remote areas. The use of a participant action research process will ensure that the educational tool will be owned by Aboriginal and Torres Strait Islander People and health-care professionals. PMID- 11421964 TI - Fractures of the femoral neck in Australian Aboriginals and Torres Strait Islanders. AB - The objective was to study patients of Aboriginal and Torres Strait Islander origin who were admitted to Cairns Base Hospital with the diagnosis of femoral neck fracture. An analysis of all 232 admissions with this diagnosis between November 1997 and July 2000 was carried out. Information was gathered from data accumulated on the Clinical Pathways database; other local data was also considered. Patients registered as being of Aboriginal and Torres Strait Islander origin have a lower incidence of these fractures than might be expected on an overall population basis, but similar rates on age-standardised data. The female age profile is substantially older than the female non-indigenous osteoporotic fracture group. Indigenous females develop osteoporotic type fractures of the femoral neck at a later age than do non-indigenous females. This may reflect a genetic difference in bone mineral density or a healthy lifestyle in earlier days. Further research is suggested. PMID- 11421965 TI - Pancreatitis in remote Australia: an indigenous perspective. AB - In Australia, pancreatitis has been associated with gallstones in approximately 35% of cases and alcohol in approximately 25% of cases. The present study was performed to analyse the aetiology of pancreatitis in patients admitted to a remote Australian hospital that has a population base comprising both indigenous and non-indigenous people. A retrospective study was conducted of all cases of pancreatitis admitted to the Alice Springs Hospital from August 1993 to August 1999. One hundred and two patient records were examined, and 77% of these patients were Aboriginal. Alcohol was the most common aetiological factor (64%), with gallstone disease being the next most common. Aboriginal patients with pancreatitis were younger than non-Aboriginal patients ( P = 0.0304) and were also more likely to have alcohol as their cause of pancreatitis ( P = 0.035), especially if they were male. In males, alcohol was implicated in at least 70% of cases. In females, the causes were gallstone disease (41%) and alcohol (49%). Patients with gallstone disease were significantly older than patients with other causes of pancreatitis ( P = 0.000). There was an overall 30% increase in admission rate per year, which was primarily due to an increase in the number of Aboriginal admissions (36% per year) ( P = 0.000). The present study highlights a difference in the aetiology of pancreatitis in Aboriginal and non-Aboriginal people from a remote Australian population. PMID- 11421966 TI - Remote area aboriginal health services managers: key practice challenges. AB - The following reflections on the author's management practice are based on the text of an address given by the author at the 1999 International Conference of the Royal Australasian College of Medical Administrators in Sydney. These reflections arise out of the author's experience for the past 5 years as manager of Nganampa Health Council, an Aboriginal community-controlled health organisation located in the remote north-west of South Australia. Nganampa Health Council is a large regional service with a national reputation for clinical and administrative excellence. It has several leading-edge health programs, which provide an exemplar for other remote health services across Australia. The author discusses three generic key management issues that remote health services managers typically encounter and argues that services are likely to be most effective when resources are applied in a focused and strategic manner and when management practices that are pragmatic and culturally appropriate are adopted. PMID- 11421967 TI - Why revised standards of practice will not be enough. PMID- 11421968 TI - The emancipation of nursing practice: applying anti-psychiatry to the therapeutic community. AB - This paper raises issues about the process and conduct of clinical relationships with people diagnosed as mentally ill who live in therapeutic communities. This clinical work is of particular importance in the late 1990s due to the changing socio-cultural climate of interaction with people living with mental illness. This climate has a focus of care on recovery in the community and not on long term hospitalization. The paper takes the position of anti-psychiatry as a preferred model of intervention because it is person and not diagnosis oriented. The nature of the therapeutic community is explored in relation to its importance in the context of destigmatizing mental illness, its structure, and in its ability to empower the person from a philosophically driven and experiential perspective. PMID- 11421969 TI - Best practice in psychogeriatric care. AB - This paper presents a best practice model for psychogeriatric care. Best practice is becoming one of the most common expressions used in the area of health care, and is often referred to in government reports and documents. The definition of 'best practice', however, is still evolving. What then, is best practice? And how can the principles of best practice be applied and integrated into the clinical speciality of psychogeriatrics? The article emphasizes the importance of evidence based interventions and the need to focus on the pragmatic aspects of providing best practice in the clinical area of psychogeriatrics; that is, what works best in practice? The position taken by the authors of this paper is that the conceptualization of a best practice model in psychogeriatrics is necessary in order to describe and explain the different components involved in the service provided. This conceptualization also communicates and articulates the role of the major stakeholders, and the key players in the achievement of best practice. A psychogeriatric service may become more coherent, more goal-orientated and more efficient if a model is utilized. This paper outlines a proposed model of best practice in psychogeriatrics, and discusses the potential implications for achieving desirable clinical outcomes. PMID- 11421970 TI - Dissociative identity disorder and the nurse-patient relationship in the acute care setting: an action research study. AB - This paper presents the results of an action research study into the acute care experience of Dissociative Identity Disorder. The study, which was grounded in principles of critical social science, utilized focus group interviews and narrative construction. Nurses and patients are under-represented in all clinical evaluation and their voices need to be heard if services are to be truly collaborative. Findings of the study extend intrapsychic theories of trauma to emphasize the interpersonal relationship between nurse and person who can work together to facilitate recovery from trauma, make connections both intra and interpersonally and build resilience. PMID- 11421971 TI - Australian Indigenous mental health. AB - Understanding the complexity of another culture's health concerns is fraught with difficulty, yet 'ways forward' abound. Many researchers, including Indigenous people, have recorded cultural understandings of health, and made recommendations that have influenced the planning of Indigenous peoples' mental health care. Indeed, there is anticipation with vision for the future. Australian Indigenous people have suffered many losses, which have resulted in much social unrest, and mental and spiritual sorrow. The difficulty of belonging and adjusting to two different cultural contexts has led to particular physical health and mental health concerns for some. Health for Indigenous people is viewed within a holistic and community lifestyle framework, which is related to both past and present issues, and it is not necessarily individualized or compartmentalized. A closer liaison between the health traditions of both cultures, working together with education, good will and understanding of each other's health business, and working together within mainstream health services may assist with healing, reconciliation and improved Aboriginal holistic health. PMID- 11421972 TI - Mental health nurses' beliefs about likely outcomes for people with schizophrenia or depression: a comparison with the public and other healthcare professionals. AB - This paper explores mental health nurses' beliefs about prognosis, long-term outcomes and discrimination in response to vignettes describing a person with either schizophrenia or depression. Mental health nurses were added to previous research describing the beliefs of professionals (psychiatrists, general practitioners and clinical psychologists) and the public. This study used a self- completion, postal survey procedure. Mental health nurses tended to be more negative than the community but more positive than the other professional groups (particularly the medical professions) about long-term outcomes and prognosis. Nurses were similar to the medical professional groups in believing that considerable discrimination occurs within the community. Given the potential impact on consumers and the public, all practitioners need to be aware of their own beliefs and those of their professional colleagues. PMID- 11421973 TI - The relationship between emotions and stress among mental health nurses. AB - This paper presents findings from an exploratory correlational study that examined the relationships between Emotional Competency, Trait Affectivity, Stress and Experienced Emotions among 43 mental health nurses in Australian regional hospitals. A significant relationship was found between Emotional Competency and Personal Self-doubt in male nurses only; however, no association was found between Emotional Competency and Experienced Emotions. Trait Affectivity was found to be associated with Experienced Emotions but not Stress. Gender differences were found in Trait Affectivity and Experienced Emotions. The results of the study have implications for the retention of mental health nurses in their profession. PMID- 11421974 TI - Re-awakenings? A discourse analysis of the recovery from schizophrenia after medication change. AB - This paper explores the construction of recovery from schizophrenia after medication change through the analysis of people living with schizophrenia. The study is framed by a discourse analysis which assumes that the language used to discuss schizophrenia and its treatment by medication is imbued with the power relations of mental health. The analysis uses research literature, pharmaceutical literature and previous studies of schizophrenia as the discursive background that frames how recovery can be talked about. The discussion highlights how the discourses of medical science construct recovery as a linear event that silences the embodiment of schizophrenia. People with schizophrenia refuse this construction, through finding a 'niche' for themselves. In conclusion, the paper suggests how such analysis opens up for exploration of the silencing of 'insanity', and establishes a beginning dialogue with people who live with the continuing presence of schizophrenia. PMID- 11421975 TI - Survey of stakeholders' opinions of community psychiatric nursing services. AB - The paper describes a telephone survey of the views of representatives of users, purchasers and providers of community psychiatric nursing services on services in Scotland. The telephone interview protocol was based on seven themes derived from interviewing 63 users, purchasers and providers throughout the UK. Descriptive statistics were used to present the responses in the survey to the forced choice questions. Responses to the open ended questions were coded, categorized and computed. The paper describes the responses by the major stakeholders to each of the themes. Overall, the quality of the existing services was placed in the 'good' category. However, criticism was directed at the insufficient numbers of Community Psychiatric Nurses in the service, the inadequate amount of time spent with clients and their poorly organized method of working. Other findings and stakeholders' recommendations are discussed. PMID- 11421976 TI - Negative attitudes towards clients with drug and alcohol related problems: finding the elusive solution. AB - The difficulties experienced by nurses in providing care to clients with drug and alcohol related problems within general hospital settings have been clearly identified in the literature. Despite this, the problem continues with little or no evidence of improvement. This paper reports the results of a research project undertaken in a large private hospital in metropolitan Melbourne, Victoria, Australia. The study sought to ascertain whether a difference in attitudes, confidence and perceived knowledge related to the care of clients with drug and alcohol problems would be evident between those nurses who used consultation and liaison services from a specialist drug and alcohol unit and those who did not. A questionnaire was administered to 200 nurses, with a 53% completion rate (n = 106). The results showed very little difference between the groups with the exception of the perceived knowledge category which indicated a statistically significant difference. PMID- 11421977 TI - A consumer's perspective of a suicide intervention programme. AB - The objective of this study was to assess the impact of a suicide intervention program from a consumer perspective. Self-administered questionnaires were distributed to consumers who had been referred to a suicide intervention counsellor in the 2-year period of the programme in rural southwest Western Australia. Three-quarters of respondents were positive about their experience with the service, with half of the respondents no longer having thoughts of suicide and only 20% of all respondents reporting having attempted deliberate self-harm postcounselling. Reported suicidal ideation and attempted self-harm were much higher in the dissatisfied group. Dissatisfaction of respondents stemmed from the history of their treatment and 'the hassle created by the many systems for them to access care'. However, the overall outcome of this study is that, from the consumer's perspective, a high intensity approach to suicide intervention resolved or improved the presenting problem and their ability to deal with it. PMID- 11421978 TI - Professional convergence in forensic practice. AB - This paper outlines the development and convergence of forensic science and secure psychiatric services in the UK, locating the professionalization of forensic nursing within a complex web of political, economic, and ideological structures. It is suggested that a stagnation of the therapeutic enterprise in high and medium security provision has witnessed an intrusion of medical power into the societal body. Expanding technologies of control and surveillance are discussed in relation to the move from modernity to postmodernity and the ongoing dynamic of medicalized offending. Four aspects of globalization are identified as impacting upon the organization and application of forensic practice: (i) organized capitalism and the exhaustion of the welfare state; (ii) security versus danger and trust versus risk; (iii) science as a meta-language; and (iv) foreclosure as a mechanism of censorship. Finally, as a challenge for the profession, some predictions are offered about the future directions or demise of forensic nursing. PMID- 11421979 TI - Undergraduate nursing students' views of mental health nursing in the forensic environment. AB - The literature clearly demonstrates that mental health nursing tends to be viewed negatively by undergraduate nursing students. While positive clinical experiences have been found to encourage more favourable attitudes towards mental health nursing, suitable placements are becoming scarce. An evaluation of clinical placements at the Victorian Institute of Forensic Mental Health (VIFMH) was undertaken to determine whether appropriate learning opportunities were offered, and identify the impact of the placement on students' attitudes to mental health nursing. The results suggest that VIFMH provides valuable learning experiences to increase students' understanding of mental health nursing and is effective in producing more positive attitudes towards this area of practice. PMID- 11421995 TI - Interim analyses and sequential designs in phase III studies. AB - Recruitment of patients to a clinical trial usually occurs over a period of time, resulting in the steady accumulation of data throughout the trial's duration. Yet, according to traditional statistical methods, the sample size of the trial should be determined in advance, and data collected on all subjects before analysis proceeds. For ethical and economic reasons, the technique of sequential testing has been developed to enable the examination of data at a series of interim analyses. The aim is to stop recruitment to the study as soon as there is sufficient evidence to reach a firm conclusion. In this paper we present the advantages and disadvantages of conducting interim analyses in phase III clinical trials, together with the key steps to enable the successful implementation of sequential methods in this setting. Examples are given of completed trials, which have been carried out sequentially, and references to relevant literature and software are provided. PMID- 11421996 TI - Beclomethasone dipropionate: absolute bioavailability, pharmacokinetics and metabolism following intravenous, oral, intranasal and inhaled administration in man. AB - AIMS: To assess the absolute bioavailability, pharmacokinetics and metabolism of beclomethasone dipropionate (BDP) in man following intravenous, oral, intranasal and inhaled administration. METHODS: Twelve healthy subjects participated in this seven-way cross-over study where BDP was administered via the following routes: intravenous infusion (1000 microg), oral (4000 microg, aqueous suspension), intranasal (1344 microg, aqueous nasal spray) and inhaled (1000 microg ex-valve, metered dose inhaler). The contribution of the lung, nose and gut to the systemic exposure was assessed by repeating the inhaled, intranasal and oral dosing arms together with activated charcoal, to block oral absorption. Blood samples were collected for 24 h postdose for the measurement of BDP, beclomethasone-17 monopropionate (B-17-MP) and beclomethasone (BOH) in plasma by liquid chromatography tandem mass spectrometry. RESULTS: Intravenous administration of BDP (mean CL 150 l h-1, Vss 20 l, t(1/2) 0.5 h) was associated with rapid conversion to B-17-MP which was eliminated more slowly (t1/2 2.7 h). In estimating the parameters for B-17-MP (mean CL 120 l h-1, Vss 424 l) complete conversion of BDP to B-17-MP was assumed. The resultant plasma concentrations of BOH were low and transient. BDP was not detected in plasma following oral or intranasal dosing. The mean absolute bioavailability (%F, 90% CI; nominal doses) of inhaled BDP was 2% (1-4%) and not reduced by coadministration of charcoal. The mean percentage F of the active metabolite B-17-MP was 41% (31-54%), 44% (34-58%) and 62% (47-82%) for oral, intranasal and inhaled dosing without charcoal, respectively. The corresponding estimates of nasal and lung absorption, based on the coadministration of charcoal, were < 1% and 36% (27-47%), respectively. CONCLUSIONS: Unchanged BDP has negligible oral and intranasal bioavailability with limited absorption following inhaled dosing due to extensive (95%) presystemic conversion of BDP to B-17-MP in the lung. The oral and intranasal bioavailabilities of the active metabolite B-17-MP were high and similar, but direct absorption in the nose was insignificant. The total inhaled bioavailability of B-17-MP (lung + oral) was also high (62%) and approximately 36% of this was due to pulmonary absorption. Estimates of oral bioavailability and pulmonary deposition based on total BOH were approximately half those found for B-17-MP. PMID- 11421997 TI - Influence of cirrhosis on lamotrigine pharmacokinetics. AB - AIMS: Lamotrigine, an antiepileptic drug, is cleared from the systemic circulation mainly by glucuronidation. The possibility of changes in the pharmacokinetics of lamotrigine in plasma owing to hepatic dysfunction has been evaluated. METHODS: Thirty-six subjects, including 24 patients with various degrees of liver cirrhosis and 12 healthy volunteers received a single 100 mg dose of lamotrgine. Blood samples were taken for 7 days in all subjects, except nine with severe cirrhosis, who had a 29 day blood sampling period. RESULTS: The pharmacokinetics of lamotrigine were comparable between the patients with moderate cirrhosis (corresponding to Child-Pugh grade A) and the healthy subjects. Plasma oral clearance mean ratios (90% confidence interval) in patients with severe cirrhosis without or with ascites (corresponding, respectively, to Child-Pugh grade B and C) to healthy subjects were, respectively, 60% (44%, 83%) and 36% (25%, 52%). Plasma half-life mean ratios (90% confidence interval) in these two patient groups to healthy subjects were, respectively, 204% (149%, 278%) and 287% (202%, 408%). CONCLUSIONS: Lamotrigine administered as a single oral dose of 100 mg was well tolerated in all groups. Initial, escalation and maintenance doses should generally be reduced by approximately 50 or 75% in patients with Child-Pugh Grade B or C cirrhosis. Escalation and maintenance doses should be adjusted according to clinical response. PMID- 11421998 TI - Dopamine D2-like receptors and amino acid-induced glomerular hyperfiltration in humans. AB - AIMS: In rodents, blockade of dopamine D2-like receptors abolishes both the physiological increase in glomerular filtration rate (GFR) induced by amino acids and the pathological hyperfiltration in experimental diabetes mellitus. This study addressed the contribution of dopamine D2-like receptors to changes in renal haemodynamics after amino acid infusion in humans. METHODS: Twelve healthy volunteers participated in this double-blind, randomized, cross-over study. GFR and renal blood flow (RPF) were assessed by renal clearance of inulin and p aminohippuric acid (PAH), respectively. Following infusion of 0.45% saline at baseline, an electrolyte-balanced solution of mixed amino acids (10%) was infused. Prior to the experiments, the subjects received orally either placebo, or sulpiride (10 mg kg-1), a centrally and peripherally acting D2-like receptor antagonist, or domperidone (1 mg kg-1) which affects only peripheral D2-like receptors. RESULTS: In the placebo series, amino acid infusion significantly increased GFR and RPF by up to 15.8 +/- 5.3% and 14.4 +/- 6.1%, respectively, while mean blood pressure and heart rate remained unchanged. Pretreatment with domperidone only marginally altered the renal response to amino acids (maximal increase by 13.2 +/- 5.6 and 11.9 +/- 4.0% in GFR and RPF, respectively), while sulpiride completely abolished the renal haemodynamic changes induced by amino acids. Total and fractional urinary sodium excretion as well as urinary osmolality were similar at baseline and increased in response to amino acids, to the same extent, in all series. No changes in renal dopamine excretion occurred. CONCLUSION: The results indicate that in man dopamine D2-like receptors are involved in the renal haemodynamic response to amino acid infusion. Whether dopamine D2-like receptor blockade diminishes glomerular hyperfiltration in pathological states requires clinical investigations. PMID- 11421999 TI - Concentration-effect relationships of two infusion rates of the imidazoline antihypertensive agent rilmenidine for blood pressure and development of side effects in healthy subjects. AB - AIMS: The aim of this study was to compare the effect profiles of i.v. administered rilmenidine aimed at the same target plasma concentration, but attained with different rates. METHODS: The study was placebo-controlled, randomized, double-blind, double-dummy, three-way, cross-over in nine healthy volunteers. All subjects randomly received either a fast infusion, a slow infusion or a placebo infusion. The target concentration was set at 8 ng ml-1 with a fivefold difference in the time to reach the maximum concentration. Saccadic eye movements, electroencephalography (EEG), blood pressure and heart rate were measured every half hour. Drug plasma concentrations, adverse events, salivary flow and visual analogue scales were obtained every hour. RESULTS: Changes in systolic/diastolic blood pressure at the end of the infusion were (mean +/- s.d.) 13.3 +/- 16.4/7.9 +/- 7.5 mmHg with the fast infusion and 16.3 +/ 12.7/10.2 +/- 7.9 mmHg during the slow infusion, compared with 0.0 +/- 13.2/1.3 +/- 8.3 mmHg for the placebo occasion. Decrease of saccadic peak velocity (SPV) at the end of the infusion was 18.0 +/- 5.2% during the fast infusion, 23.0 +/- 7.0% with the slow infusion, and 6.1 +/- 5.2% with placebo. PK/PD analysis showed similar concentration-dependent linear reduction in SPV and blood pressure up to the maximum observed rilmenidine plasma level for both the fast and the slow infusion. CONCLUSIONS: The rate of infusion of rilmenidine in healthy volunteers does not influence the PK/PD relationship of saccadic eye movements and blood pressure up to the maximum observed rilmenidine plasma concentrations. Rilmenidine causes clear concentration-dependent blood pressure reductions in healthy volunteers. PMID- 11422000 TI - The effect of anastrozole on the single-dose pharmacokinetics and anticoagulant activity of warfarin in healthy volunteers. AB - AIMS: The aims of this study were to determine the effects of the nonsteroidal, selective aromatase inhibitor, anastrozole, at steady-state concentrations, on the pharmacokinetics and pharmacodynamics of warfarin, and to assess whether or not anastrozole alone has any anticoagulant activity. METHODS: This was a randomized, double-blind, placebo-controlled, two-way crossover trial conducted at a single centre. The study comprised two treatment periods of 11 days, separated by a 3 week washout. Healthy male volunteers (n = 16, median age 28.5 years) were randomized to receive either anastrozole (7 mg loading dose on day 1, followed by 1 mg daily on days 2-11) in the first treatment period and placebo in the second treatment period, or vice versa. In addition to their randomized treatment, all volunteers received a single dose of 25 mg warfarin on day 3 of each treatment period. Blood samples for pharmacokinetic and pharmacodynamic assessment were taken at frequent intervals during each treatment period. The safety of volunteers was monitored throughout the study. RESULTS: Administration of anastrozole resulted in no clinically significant changes in the pharmacokinetics of either R- or S-warfarin compared with placebo for AUC (ng ml 1 h) (glsmean, R-warfarin; anastrozole 93619.9, placebo 91127.91, 95%CI 0.988 1.068; S-warfarin; anastrozole 57129.21, placebo 55676.34, 95%CI 0.979-1.076), CL/F (ml min-1) (glsmean, R-warfarin; anastrozole 2.23, placebo 2.29, 95%CI 0.937 1.012; S-warfarin; anastrozole 3.65, placebo 3.74, 95%CI 0.929-1.021) and t1/2 (h) (lsmean, R-warfarin; anastrozole 55.40, placebo 55.15, 95%CI-2.083-2.592; S warfarin; anastrozole 39.38, placebo 40.98, 95%CI-6.189-2.996). In addition, anastrozole had no clinically significant effect on the pharmacodynamic effects of warfarin, as assessed 240 h after warfarin dosing by measurement of prothrombin time (s) (glsmean, anastrozole 11.56, placebo 11.31, 95%CI 0.987 1.059), thrombin time (s) (glsmean, anastrozole 19.06, placebo 18.75, 95%CI 0.980 1.054) activated partial thromboplastin time (s) (glsmean, anastrozole 29.94, placebo 29.74, 95%CI 0.968-1.047) and factor VII (%) (glsmean, anastrozole 97.81, placebo 107.26, 95%CI 0.821-1.012). Anastrozole alone had no effect on these indicators of the clotting process. CONCLUSIONS: Overall, there was no evidence to suggest that anastrozole has any clinically relevant effects on the pharmacokinetics of warfarin. Anastrozole had no effect on clotting mechanisms or on the pharmacodynamic activity of warfarin, as assessed by prothrombin time, thrombin time, activated partial thromboplastin time, and factor VII. PMID- 11422001 TI - Effect of MAO-A inhibition on the pharmacokinetics of almotriptan, an antimigraine agent in humans. AB - AIMS: To assess the effect of a reversible MAO-A inhibitor, moclobemide, on the single-dose pharmacokinetics of almotriptan and assess the clinical consequences of any interaction. METHODS: Twelve healthy volunteers received the following treatments in a randomized, open-label, two-way crossover design (with a 1 week washout between treatments): (A) one 150 mg moclobemide tablet every 12 h for 8 days and one 12.5 mg almotriptan tablet on the morning of day 8; and (B) one 12.5 mg almotriptan tablet on day 8. Plasma almotriptan was quantified by h.p.l.c.-MS MS, while urinary concentrations were measured by h.p.l.c.-u.v. Vital signs, ECGs, and adverse events were evaluated after almotriptan administration. Treatment effects on pharmacokinetics and vital signs were assessed by analysis of variance. RESULTS: Mean almotriptan AUC was higher (483 +/- 99.9 vs 352 +/- 75.4 ng ml-1 h, P = 0.0001) and oral clearance was lower (26.6 +/- 4.00 vs 36.6 +/- 5.89 l h-1, P = 0.0001) when almotriptan was administered with moclobemide. Mean half-life was longer (4.22 +/- 0.78 vs 3.41 +/- 0.45 h, P = 0.0002) after coadministration with moclobemide. Renal clearance of almotriptan was unaffected by moclobemide. No serious adverse events occurred and no clinically significant vital sign changes were observed. CONCLUSIONS: Moclobemide increased plasma concentrations of almotriptan on average by 37%, but the combined administration of these two compounds was well tolerated. The degree of interaction was much less than that seen previously for sumatriptan or zolmitriptan given with moclobemide. PMID- 11422002 TI - Effect of itraconazole on the pharmacokinetics of prednisolone and methylprednisolone and cortisol secretion in healthy subjects. AB - AIMS: Itraconazole is a potent inhibitor of CYP3A4 activity and is often used in combination with corticosteroids. Since the latter are partly metabolized by CYP3A4, we studied the interaction between itraconazole, prednisone and methylprednisolone in healthy male subjects. METHODS: The effects of 4 days administration of oral itraconazole (400 mg on the first day then 200 mg day-1 for 3 days) on the pharmacokinetics of prednisolone after a single oral dose of prednisone (60 mg) and the pharmacokinetics of methylprednisolone after single oral dose of methylprednisolone (48 mg) were studied in 14 healthy male subjects in a two-period cross-over trial. Plasma cortisol concentrations were determined as a pharmacodynamic index. RESULTS: Itraconazole increased the mean area under the methylprednisolone concentration-time curve from 2773 ng ml-1 h to 7011 ng ml 1 h (P < 0.001) and the elimination half-life from 3.2 h to 5.5 h (P < 0.001). The pharmacokinetics of prednisolone were unchanged. Cortisol concentrations at 24 h were lower after administration of methylprednisolone with itraconazole than after methylprednisolone alone (24 ng ml-1 vs 109 ng ml-1, P < 0.001). CONCLUSIONS: Itraconazole increased methylprednisolone concentrations markedly with enhanced suppression of endogenous cortisol secretion, but had no effect on prednisolone pharmacokinetics. The pharmacokinetic interaction between methylprednisolone and itraconazole is probably related to inhibition of hepatic CYP3A4 activity by itraconazole. PMID- 11422003 TI - Enterocytic CYP3A4 in a paediatric population: developmental changes and the effect of coeliac disease and cystic fibrosis. AB - AIMS: To investigate the effects of age and disease states on the expression and activity of intestinal CYP3A4 in a paediatric population. METHODS: Duodenal biopsies and surgical sections were collected from 104 paediatric patients (age range 2 weeks to 17 years) and from 11 foetuses. An S9 fraction was prepared in each case. CYP3A4 expression was assessed by Western blotting and by immunohistochemistry; activity was measured by the rate of formation of 6beta hydroxytestosterone from testosterone. Villin expression was used as a marker of enterocyte harvest to normalize CYP3A4 expression and activity data. RESULTS: In the 74 histologically normal paediatric biopsies there were statistically significant increases in CYP3A4 expression (r2 = 0.19, P = 0.001) and activity (r2 = 0.17, P = 0.02) with age. CYP3A4 was practically absent in fetal duodenum and was expressed at relatively low levels in neonates (P < 0.05 between neonates and children > 5 years). Active coeliac disease resulted in significant (P < 0.001) decreases in CYP3A4 expression and activity. CONCLUSIONS: Duodenal CYP3A4 is present at significantly lower levels in neonates and in patients with active coeliac disease. This may have clinical significance with respect to the oral bioavailability of CYP3A4 substrates. PMID- 11422004 TI - Inhibitory effects of verapamil and diltiazem on simvastatin metabolism in human liver microsomes. AB - AIMS: To determine the effects of verapamil and diltiazem on simvastatin metabolism in human liver microsomes and to compare their inhibitory potencies and CYP3A4 inactivation parameters with those reported previously for mibefradil. METHODS: Simvastatin metabolism was investigated in human liver microsomes in the presence and absence of verapamil or diltiazem (0.1-250 microM). Kinetics of CYP3A4 inactivation by verapamil and diltiazem were determined using testosterone as the substrate. RESULTS: When verapamil was coincubated with simvastatin, IC50 values ranged from 23 to 26 microM for all major metabolites. The IC50 values ranged from 4.8 to 5.6 microM on preincubation of verapamil for 30 min in the presence of an NADPH-generating system. Corresponding IC50 values for diltiazem ranged from 110-127 microM and from 21-27 microM, respectively. Verapamil and diltiazem inhibited testosterone 6beta-hydroxylation in a time- and concentration dependent manner, key features of mechanism-based inactivation. Values for the inactivation parameters kinact and KI were 0.15 +/- 0.04 min-1 (mean +/- s.d.) and 2.9 +/- 0.6 microM, respectively, for verapamil and 0.07 +/- 0.01 min-1 and 3.3 +/- 1.5 microM, respectively, for diltiazem. CONCLUSIONS: The IC50 values for coincubation of verapamil and diltiazem were 46- and 220-fold higher, respectively, than those reported previously for mibefradil, and 16- and 71-fold higher, respectively, for preincubation. Thus, the results of this study suggest that verapamil and diltiazem are less likely than mibefradil to cause acute drug interactions with simvastatin in vivo. However, verapamil and diltiazem are moderate mechanism-based inhibitors of CYP3A4 and therefore may still cause significant inhibition of simvastatin metabolism in vivo during chronic therapy. PMID- 11422005 TI - Mephenytoin as a probe for CYP2C19 phenotyping:effect of sample storage, intra individual reproducibility and occurrence of adverse events. AB - AIMS: To further evaluate mephenytoin as a probe for CYP2C19 phenotyping. METHODS: Healthy subjects (n = 2638) were phenotyped using the urinary (S) mephenytoin to (R)-mephenytoin ratio. This method was evaluated for (a) the stability of the S/R-ratio following sample storage, (b) the intraindividual reproducibility of the ratio, and (c) the occurrence of adverse events. RESULTS: After prolonged storage, the S/R-ratio of samples from extensive metabolisers (EM) increased up to 85%. In 1.5% of the cases (1 out 66), this led to incorrect classification of phenotype. In EMs, but not in poor metabolisers (PMs), the S/R ratio increased after acid treatment. The intraindividual reproducibility of the mephenytoin phenotyping procedure was 28%. No major side-effects were observed and there was no relationship between the incidence of side-effects and the phenotype of the subject. CONCLUSIONS: After prolonged storage the S/R-ratio significantly increased in EMs and, although low, the risk of incorrect classification should not be ignored. Our data support the use of mephenytoin as a safe drug for CYP2C19 phenotyping. PMID- 11422006 TI - Frequencies of thiopurine S-methyltransferase mutant alleles (TPMT*2, *3A, *3B and *3C) in 151 healthy Japanese subjects and the inheritance of TPMT*3C in the family of a propositus. AB - AIMS: To determine the frequencies of four thiopurine S-methyltransferase (TPMT) mutant alleles, TPMT*2, *3A, *3B and *3C in a normal Japanese population. METHODS: Genotypes were determined in 151 Japanese subjects and in six family members of a propositus using polymerase chain reaction (PCR)-restriction fragment length polymorphism and allele-specific PCR assays. RESULTS: Only one TPMT*3C heterozygote was identified (gene frequency 0.3%). TPMT*2, *3A and *3B were not detected. In addition, TPMT*3C was found to have been inherited from the mother and passed on to the son of the propositus. CONCLUSIONS: TPMT*3C appears to be most prevalent among the known mutant allele of TPMT in a Japanese population which may have some relevance for the treatment of Japanese patients with thiopurine drugs. PMID- 11422007 TI - Quinapril and its metabolite quinaprilat in human milk. AB - AIMS: To measure the milk to plasma ratio (M/P) of quinapril and its active metabolite quinaprilat in lactating mothers and to assess likely infant exposure. METHODS: A single dose of quinapril 20 mg was administered to six healthy mothers who had been breastfeeding their infants for at least 2 weeks. Blood was sampled for the measurement of quinapril and quinaprilat at 0, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, 16 and 24 h. Milk was collected for measurement of quinapril and quinaprilat concentrations over the periods -4-0, 0-4, 4-8, 8-12, 12-18, 18-24 h. The areas under the plasma and milk concentration-time curves were estimated and an M/P ratio derived for both quinapril and quinaprilat. RESULTS: The M/P ratio for quinapril was 0.12 (95% CI 0.09,0.14). No quinapril was detected in milk after 4 h. No quinaprilat was detected in any of the milk samples. The estimated 'dose' of quinapril that would be received by the infant was 1.6% (95% CI 1.0,2.2) of the maternal dose, adjusted for respective weights. CONCLUSIONS: Quinapril appears to be 'safe' during breastfeeding according to conventional criteria, although as always, the risk:benefit ratio should be considered when it is to be given to a nursing mother. PMID- 11422008 TI - Influence of ACE-inhibition on salt-mediated worsening of pulmonary gas exchange in heart failure. AB - AIMS: In congestive heart failure (CHF), pulmonary gas exchange, as evaluated by carbon monoxide diffusion (DLCO), is impaired. ACE-inhibition improves DLCO. Infusion of saline worsens DLCO, because of upregulated sodium and water transport to the alveolar interstitium, which thickens the alveolar-capillary interface and lengthens the diffusion path for gas exchange. We investigated whether enalapril can readjust the capillary permeability to sodium. METHODS: In 10 NYHA class II-III CHF patients, we measured DLCO, its two subcomponents (VC, capillary blood volume available for gas exchange, and DM, alveolar-capillary membrane diffusion), left and right ventricular filling pressures, plasma noradrenaline, aldosterone and renin activity, at baseline and following saline infusion in the main pulmonary artery stem, before and after 1 week enalapril treatment (20 mg daily). RESULTS: Saline (150 ml) significantly reduced DLCO ( 9.1%) and DM (-9.8%) and augmented VC (+ 10.7%). Responses to 750 ml saline were somewhat greater and qualitatively similar. Enalapril produced a significant improvement of DLCO and DM at rest as well as after saline, that was not associated with variations in ventricular filling pressures, cardiac output and left ventricular ejection fraction, and was not accounted for by humoral changes. CONCLUSIONS: In CHF, ACE-inhibition attenuates the deterioration of pulmonary gas transfer produced by saline infusion, suggesting an ability to readjust the upregulated sodium transport across the pulmonary microvascular endothelium. PMID- 11422009 TI - Hepatitis B vaccine and first episodes of central nervous system demyelinating disorders: a comparison between reported and expected number of cases. PMID- 11422010 TI - Pulse wave analysis. PMID- 11422011 TI - Stopping rules for phase II studies. AB - This paper, the second in a series of three papers concerned with the statistical aspects of interim analyses in clinical trials, is concerned with stopping rules in phase II clinical trials. Phase II trials are generally small-scale studies, and may include one or more experimental treatments with or without a control. A common feature is that the results primarily determine the course of further clinical evaluation of a treatment rather than providing definitive evidence of treatment efficacy. This means that there is more flexibility available in the design and analysis of such studies than in phase III trials. This has led to a range of different approaches being taken to the statistical design of stopping rules for such trials. This paper briefly describes and compares the different approaches. In most cases the stopping rules can be described and implemented easily without knowledge of the detailed statistical and computational methods used to obtain the rules. PMID- 11422012 TI - 6-Thioguanine in children with acute lymphoblastic leukaemia: influence of food on parent drug pharmacokinetics and 6-thioguanine nucleotide concentrations. AB - AIMS: Since relatively little is known about the pharmacokinetics of 6 thioguanine (6TG) in children receiving 6-thioguanine for maintenance therapy of acute lymphoblastic leukaemia (ALL), we studied plasma drug concentrations under standardized conditions and investigated the effect of food on parent drug pharmacokinetics and the accumulation of the active metabolites 6-thioguanine nucleotides (6-TGNs) in red cells. METHODS: Single oral doses of 40 mg of 6-TG were administered both in the fasting and fed state to children with ALL. Pharmacokinetic sampling was performed up to 6 h post dose. Daily oral doses of 40 mg m(-2) of 6-TG were administered both fasting and after food over two 4 week periods. Twice weekly samples were taken for metabolite concentrations. The study design was cross-over with each child receiving dosing in either fasted or after food over a 4 week period in each phase. RESULTS: Eleven patients were studied. A wide interindividual variation in Cmax (median 313 pmol ml(-1), range 51-737) and AUC (median 586 pmol ml(-1) h, range 156-1306) was observed in the fasted state. Concomitant food administration resulted in a significant reduction in Cmax (median 71 vs 313 pmol ml(-1), P = 0.006, CI from 36 to 426), AUC (median 200 vs 586 pmol ml(-1) h, P = 0.006, 95% CI from 109 to 692), and time to reach Cmax (median 1.5 vs 3 h, P = 0.013, 95% CI from 0.74 to 2.73). There was no difference in the steady state concentration of red cell 6-TGNs observed after a 4 week period of 6-TG administered fasting or after food. CONCLUSIONS: Children with ALL demonstrate significant interindividual variation in 6-TG pharmacokinetics. Although there would appear to be a reduction in parent drug Cmax and AUC with food there was no difference in 6-TGN concentrations after 4 weeks of 6-TG. Taking the drug on an empty stomach may not be necessary. PMID- 11422013 TI - Oral bioavailability of dihydroartemisinin in Vietnamese volunteers and in patients with falciparum malaria. AB - AIMS: To obtain comprehensive bioavailability data for artesunate (ARTS) and its active metabolite dihydroartemisinin (DHA) following their separate oral administration to Vietnamese volunteers and to patients with acute, uncomplicated falciparum malaria. METHODS: Volunteers were randomized to receive either i.v. ARTS (120 mg) followed by oral ARTS (150 mg) 8 h later (Group 1, n = 10), or i.v. ARTS (120 mg) followed by oral DHA (120 mg) 8 h later. Patients, also received oral ARTS (150 mg; Group 3, n = 8) or DHA (120 mg; Group 2, n = 7), in a randomized cross-over study design. Multiple blood samples were collected after each administration and plasma ARTS and/or DHA concentrations were determined by h.p.l.c. Pharmacokinetic descriptors were obtained from noncompartmental analysis and bioavailability was calculated from AUC data. In the patients, the time to 50% parasite clearance (PCT50) and fever clearance time (FCT) also were measured. RESULTS: In Group 1 (volunteers), the mean (95% CI) absolute bioavailability of oral ARTS was 80% (62,98%), while in Group 2 (volunteers), the bioavailability of oral DHA was 45% (34,56%). In the patients (Group 3), the bioavailability of oral DHA relative to oral ARTS was 88% (49,127%). The median PCT50 and FCT were 2.3 and 28 h, respectively. CONCLUSIONS: The study shows that the absolute bioavailability of DHA was significantly lower than that for ARTS in healthy volunteers. The bioavailability of ARTS in volunteers was consistent with previous studies in patients with uncomplicated falciparum malaria. The dose normalized Cmax and AUC(0,infinity) for DHA were significantly greater in patients with falciparum malaria than in healthy volunteers. The high relative bioavailability of DHA in the patients may have been due to lower first-pass clearance. We conclude that, for the treatment of malaria, DHA is likely to be a suitable oral substitute for ARTS. Based on our mean AUC measurements, it appears that equal doses of DHA and ARTS (mg basis) should give equivalent systemic exposure to bioactive DHA in uncomplicated falciparum malaria. PMID- 11422014 TI - Simultaneous administration of a cocktail of markers to measure renal drug elimination pathways: absence of a pharmacokinetic interaction between fluconazole and sinistrin, p-aminohippuric acid and pindolol. AB - AIMS: Previous studies suggest that estimated creatinine clearance, the conventional measure of renal function, does not adequately reflect changes in renal drug handling in some patients, including the immunosuppressed. The aim of this study was to develop and validate a cocktail of markers, to be given in a single administration, capable of detecting alterations in the renal elimination pathways of glomerular filtration, tubular secretion and tubular reabsorption. METHODS: Healthy male subjects (n = 12) received intravenously infused 2500 mg sinistrin (glomerular filtration) and 440 mg p-aminohippuric acid (PAH; anion secretion), and orally administered 100 mg fluconazole (reabsorption) and 15 mg rac-pindolol (cation secretion). The potential interaction between these markers was investigated in a pharmacokinetic study where markers (M) or fluconazole (F) were administered alone or together (M + F). Validated analytical methods were used to measure plasma and urine concentrations in order to quantify the renal handling of each marker. Plasma protein binding of fluconazole was measured by ultrafiltration. All subjects had an estimated creatinine clearance within the normal range. The renal clearance of each marker (mean+/- s.d.) was calculated as the ratio of the amount excreted in urine and the area-under-the-concentration time curve. Statistical comparisons were made using a paired t-test and 95% confidence intervals were reported. RESULTS: The renal clearances of sinistrin (M: 119 +/- 31 ml min(-1); M + F: 130 +/- 40 ml min(-1); P = 0.32), PAH (M: 469 +/- 145 ml min(-1); M + F: 467 +/- 146 ml min(-1); P = 0.95), R-pindolol (M: 204 +/- 41 ml min(-1); M + F: 190 +/- 41 ml min(-1); P = 0.39; n = 11), S-pindolol (M: 225 +/- 55 ml min(-1); M + F: 209 +/- 60 ml min(-1); P = 0.27; n = 11) and fluconazole (F: 14.9 +/- 3.8 ml min(-1); M + F: 13.6 +/- 3.4 ml min(-1); P = 0.16) were similar when the markers or fluconazole were administered alone (M or F) or as a cocktail (M + F). CONCLUSIONS: This study found no interaction between markers and fluconazole in healthy male subjects, suggesting that a single administration of this cocktail of markers of different renal processes can be used to simultaneously investigate pathways of renal drug elimination. PMID- 11422015 TI - Increased oral ganciclovir bioavailability in HIV-infected patients with chronic diarrhoea and wasting syndrome--a population pharmacokinetic study. AB - AIMS: Despite a lack of data, the antiviral agent ganciclovir is not indicated in AIDS patients with diarrhoea because of its presumed poor oral bioavailability. To assess the effect of diarrhoea on ganciclovir intestinal absorption, we conducted a pharmacokinetic study in 42 HIV-infected patients categorized into three groups: A, HIV stage A and B (n = 15); B, AIDS stage C (n = 13); C, AIDS with chronic diarrhoea and wasting syndrome (n = 14). METHODS: Each patient was evaluated for nutritional (body mass index, albumin, transferrin serum levels), inflammatory (haptoglobin, orosomucoid), immunological (CD4 count, plasma viral load) and intestinal (D-xylose test, faecal fat and nitrogen output, intestinal permeability) status. Ganciclovir (1 g) was administered orally to fasted patients. Six blood samples were collected over 24 h. Serum was analysed for ganciclovir by h.p.l.c. Population pharmacokinetic analysis was performed using a nonlinear mixed effects modelling program, MP2. RESULTS: Mean intestinal permeability (lactulose/mannitol urinary ratio) was increased in group C (0.2) compared with group A (0.05) and B (0.1) patients. Drug concentration-time profiles were best described by a two-compartment model. Apparent oral clearance (CL/F) and central volume of distribution (V1/F) were influenced by clinical status (group). For groups A and B combined, final parameter estimates of CL/F and V1/F were 256 +/- 98 l h(-1) and 1320 +/- 470 l, respectively. Final parameter estimates for group C were 118 +/- 108 l h(-1) and 652 +/- 573 l for CL/F and V1/F, respectively. The 95% confidence intervals on differences between A and B combined and C were statistically significant ([ + 70, + 206] for CL/F, and [+ 314, + 1022] for V1/F). Compared with groups A and B, ganciclovir CL/F was significantly decreased in group C patients. CONCLUSIONS: AIDS patients with diarrhoea and severe disease may benefit from ganciclovir therapy, but a dose adjustment may be required according to their digestive and immunological status. PMID- 11422016 TI - Population pharmacokinetics of carbamazepine in Singapore epileptic patients. AB - AIMS: To document the population pharmacokinetics of carbamazepine in patients with epilepsy living in Singapore, the majority of whom are of Chinese origin and others of minority races. METHODS: Steady-state plasma carbamazepine concentration data were gathered during routine care from various hospitals in Singapore for patients with epilepsy. Age, body weight, gender, race, formulation and concurrent medication (for other illnesses) were the fixed effects (covariates) tested simultaneously for their influence on the population mean of carbamazepine clearance, using the nonlinear mixed-effects model, in the NONMEM program. RESULTS: No age, gender, race, or formulation-related effect was found. Body weight (W), age (A) and concurrent medication with phenobarbitone (PB) emerged as the determinants of carbamazepine clearance (CL). The final regression model for carbamazepine clearance found best to describe the data was CL = 40.7 x A(0.494) x W(-1.17) x 1.44PB where CL is in l day(-1) kg(-1), A is in years, W is in kg and PB = 0 for a patient on carbamazepine only and PB = 1 for a patient on concomitant PB. The corresponding interindividual variability (CV%) in CL, described by using an exponential model, was 21.4%, and the residual error, described by using an exponential error model, was 18.2%. Predictive performance of this population covariate model was evaluated by Bayesian forecasting in a similar, but independent cohort of patients. There was no statistically significant bias between predicted and measured plasma carbamazepine concentrations. The population mean value of carbamazepine clearance obtained was similar to that previously reported for patients with a very different ethnic (Caucasians and Blacks) or geographical background (South Africa, Europe and USA). CONCLUSIONS: The derived covariate regression model reasonably predicted concentrations in the separate validation Singapore patient data set. The correlation between carbamazepine clearance and patient-specific characteristics may thus allow dosage adjustment to be made to achieve target steady-state plasma concentrations. PMID- 11422017 TI - Platelet CD62 expression and PDGFAB secretion in patients undergoing PTCA and treatment with abciximab. AB - AIMS: To investigate a correlation of the platelet activation marker CD62 and secretion of the growth factor PDGF from platelets in coronary patients under therapy with the GPIIb/IIIa-inhibitor abciximab. METHODS: Flow cytometric assessment of fibrinogen binding (GPIIb/IIIa-binding site) and CD62 expression, as well as PDGF release of human platelets (immunoassay) and platelet aggregation with 20 microM ADP and 2 microg ml(-1) collagen were evaluated in nine patients with stable coronary artery disease. Patients were undergoing elective balloon angioplasty and were treated with aspirin (100 mg day(-1)), heparin (ACT < 220 s) and abciximab (bolus and infusion over 12 h). Blood samples were obtained before initiation of abciximab therapy (under aspirin and heparin) (I), 3 h after angioplasty under abciximab (II) and 12 h after termination of abciximab infusion (III). RESULTS: Compared with sample I before abciximab therapy, fibrinogen binding was reduced to 37% (+/- 34 s.d., P < 0.05) (II) and 55% (+/- 40 s.d., P < 0.05) (III). Reduced fibrinogen binding also led to a significant reduction of the aggregation response to ADP (down to 37% +/- 20) and collagen (down to 0%). Mean fluorescence intensity of CD62-expression was 78 units (+/- 20 s.d.) (I), 72 units (+/- 14 s.d.) (II) and 64 units (+/- 12 s.d., P < 0.05) (III). PDGF release from isolated, washed platelets was 99 (+/- 33 s.d.) ng/10(9) platelets at (I), 82 (+/- 31 s.d.) ng/10(9) platelets and 96 (+/- 30 s.d.) ng/10(9) platelets. CONCLUSIONS: The results indicate that despite a strong reduction of GPIIb/IIIa binding and platelet aggregation, CD62 as a marker of platelet secretion and the secretion product PDGF were only slightly reduced under abciximab treatment. No direct correlation between CD62 expression and PDGF release could be demonstrated. PMID- 11422018 TI - Comparison of the effects of nadolol and bisoprolol on the isoprenaline-evoked dilatation of the dorsal hand vein in man. AB - AIMS: We attempted to explore the possible differential involvement of beta adrenoceptor subtypes in the dilator response of the human dorsal hand vein to isoprenaline by examining the ability of bisoprolol, a selective beta1 adrenoceptor antagonist, and nadolol, a nonselective beta1/beta2-adrenoceptor antagonist, to antagonize the response. METHODS: Twelve healthy male volunteers participated in four weekly sessions. In the preliminary session a dose-response curve to the vasoconstrictor effect of phenylephrine was constructed and the dose producing 50-75% maximal response was determined for each individual. In each of the remaining three (treatment) sessions, nadolol (40 mg), bisoprolol (5 mg) or placebo was ingested, and isoprenaline hydrochloride (3.33-1000 ng min(-1)) was infused locally into the dorsal hand vein along with a constant dose of phenylephrine hydrochloride (to preconstrict the vein) 2 h after the ingestion of the drugs. Changes in vein diameter were monitored with the dorsal hand vein compliance technique. Subjects were allocated to treatment session according to a double-blind balanced cross-over design. Systolic and diastolic blood pressure, and heart rate were also measured. RESULTS: Isoprenaline produced dose-dependent venodilatation which was antagonized by nadolol but remained unaffected by bisoprolol (ANOVA with repeated measures: P < 0.025; Dunnett's test: placebo vs nadolol, P < 0.01; placebo vs bisoprolol, P = NS). Mean log ED50 (ng min-1) was significantly increased in the presence of nadolol and remained unchanged in the presence of bisoprolol (ANOVA, P < 0.025; Dunnett's test: placebo vs nadolol, P < 0.005; placebo vs bisoprolol, P = NS; differences between mean log ED50 [95% CI]: placebo vs bisoprolol -0.11 [-0.38, 0.16], placebo vs nadolol 0.32[0.09, 0.72], bisoprolol vs nadolol -0.43 [-0.71, -0.15]). Mean Emax did not differ in the three treatment conditions. CONCLUSIONS: The failure of bisoprolol to attenuate isoprenaline-evoked venodilatation in the human dorsal hand vein argues against the involvement of a beta1-adrenoceptor-mediated component in the isoprenaline evoked venodilatory responses. The possibility cannot be excluded that the consequences of beta1-adrenoceptor blockade by bisoprolol might have been obscured by a possible venodilator effect of bisoprolol. PMID- 11422019 TI - Pharmacokinetic interaction between mefloquine and ritonavir in healthy volunteers. AB - AIMS: To evaluate the pharmacokinetic interaction between ritonavir and mefloquine. METHODS: Healthy volunteers participated in two separate, nonfasted, three-treatment, three-period, longitudinal pharmacokinetic studies. Study 1 (12 completed): ritonavir 200 mg twice daily for 7 days, 7 day washout, mefloquine 250 mg once daily for 3 days then once weekly for 4 weeks, ritonavir restarted for 7 days simultaneously with the last mefloquine dose. Study 2 (11 completed): ritonavir 200 mg single dose, mefloquine 250 mg once daily for 3 days then once weekly for 2 weeks, ritonavir single dose repeated 2 days after the last mefloquine dose. Erythromycin breath test (ERMBT) was administered with and without drug treatments in study 2. RESULTS: Study 1: Ritonavir caused less than 7% changes with high precision (90% CIs: -12% to 11%) in overall plasma exposure (AUC(0,168 h)) and peak concentration (Cmax) of mefloquine, its two enantiomers, and carboxylic acid metabolite, and in the metabolite/mefloquine and enantiomeric AUC ratios. Mefloquine significantly decreased steady-state ritonavir plasma AUC(0,12 h) by 31%, Cmax by 36%, and predose levels by 43%, and did not affect ritonavir binding to plasma proteins. Study 2: Mefloquine did not alter single dose ritonavir pharmacokinetics. Less than 8% changes in AUC and Cmax were observed with high variability (90%CIs: -26% to 45%). Mefloquine had no effect on the ERMBT whereas ritonavir decreased activity by 98%. CONCLUSIONS: Ritonavir minimally affected mefloquine pharmacokinetics despite strong inhibition of CYP3A4 activity from a single 200 mg dose. Mefloquine had variable effects on ritonavir pharmacokinetics that were not explained by hepatic CYP3A4 activity or ritonavir protein binding. PMID- 11422020 TI - Public health impact of adverse bone effects of oral corticosteroids. AB - AIMS: The objective of this study was to estimate the number of fractures attributed to oral corticosteroid use. METHODS: Information was obtained from the General Practice Research Database which contains medical records of general practitioners in the UK. The total number of corticosteroid-related fractures during a course of treatment was estimated using the formula for attributable risk among the exposed. RESULTS: A total of 244 235 patients was prescribed an oral corticosteroid. The rate of hip fractures increased exponentially with age in both males and females. The excess number of hip fracture cases among females aged 85 years or older using 7.5 mg prednisolone per day or more was 1.4 cases per 100 patients per year. About 47% of all hip and 72% of all vertebral fractures that occurred can be attributed to oral corticosteroid use. Among 10 000 female users of higher doses, 99.7 nonvertebral, 31.6 hip and 45.8 vertebral fractures can be attributed to use of oral corticosteroids. CONCLUSIONS: The targeting of high-risk patients will be important for implementing preventative strategies in a cost-effective manner. PMID- 11422021 TI - A 6-month large-scale study into the safety of tamsulosin. AB - AIMS: Tamsulosin is an alpha1-adrenoceptor antagonist for the treatment of symptomatic benign prostatic hyperplasia with a tolerability similar to that of placebo in short-term, placebo-controlled studies with limited patient numbers. The present study was designed to test the safety of tamsulosin treatment in a large cohort of men during a prolonged period of time, particularly with regard to comedications. METHODS: A multicentre, open-label phase IIIb study with 1784 patients receiving 0.4 mg o.d. tamsulosin for 6 months was performed according to good clinical practice guidelines. The analysis was performed on an intention-to treat basis and powered to detect adverse events (AE) occurring in 0.15% of patients with 95% confidence. RESULTS: During a total drug exposure time of 811 patient years, 386 AE were recorded in 253 patients (14.2%; 95% confidence intervals [CI] 12.0-15.2%). Twenty-nine patients suffered 44 serious AE including five fatal events (CI 0.12-0.73%) due to myocardial infarction (n = 3) and to pneumonia and a car accident (one each), but all deaths were judged to be unlikely to be related to study medication. The frequency of AE in patients without any comedication (n = 1095) was 13.0% (CI 11.3-14.9%). In a logistic regression analysis beta-adrenoceptor blockers, converting enzyme inhibitors, antidiabetics and diuretics did not significantly affect the odds ratio for having AE. However, concomitant alpha-adrenoceptor antagonists (a protocol violation) and treatment with verapamil (which also has alpha-adrenoceptor antagonist activity) significantly enhanced the odds ratio for having AE to 3.87 (CI 1.52-9.85) and 3.17 (CI 1.52-6.58), respectively. Minor increases in the odds ratio, which did not reach statistical significance, were also observed for Ca2+ antagonists other than verapamil and for nitrates. CONCLUSIONS: We conclude that tamsulosin has a good safety profile relative to AE rates in the placebo arms of previous studies on tamsulosin even in the presence of most potentially complicating comedications. No major unexpected severe AE were recorded during our 6 months study. PMID- 11422022 TI - Drug therapy in the elderly: what doctors believe and patients actually do. AB - AIMS: To examine the medication adherence among old persons living in their own homes, to assess their knowledge of their medication, and to indicate target areas for intervention. METHODS: A cross-sectional study of data collected from randomly selected samples of 348 persons, aged 75 years recruited from a population-based register in the municipality of Aarhus, Denmark. Information on all drugs was collected from the subjects during a home visit, and their drug storage was examined. Information was collected from the general practitioners (GP). The measures of adherence were scores of agreements between the GPs' lists and the subjects' actual drug consumption. RESULTS: We found disagreement between the drug information collected from the study population and from the GPs: concerning drugs in 22% of the study-population, concerning doses in 71%, and concerning regimens prescribed by the GP in 66%. Twenty-four percent stated that they did not always follow prescriptions. Most of the deviations from prescriptions were toward lower doses and less frequent drug intake. The drugs most often involved in deviations were hypnotics, analgesics, bronchodilators and diuretics. Sixty percent of the participants knew the purpose of medication, and 21% knew the consequences of omission of the drugs. Less than 6% of the subjects knew about the toxic risks, side-effects, or potential drug interactions. The participants' knowledge of the drugs was positively associated with their adherence. We found a correlation between an increased number of prescribed frequency of drug intake per day and deviation from the regimen (r = 0.25, P = 0.01). There was a positive association between nonadherence and the use of three or more drugs (odds ratio (OR) 2.5; 95% confidence interval (CI) 1.5,4.1), prescriptions from more than one doctor (OR 2.5; 95% CI 1.3,4.8), and probability of dementia (OR 9.0; 95% CI 1.1,72.5). Moreover compliance aids facilitated adherence (OR 4.4; 95% CI 1.6,12.3). Persons living alone were more prone to medication errors (OR 2.0; 95% CI 1.1,3.5). CONCLUSIONS: A differentiated evaluation of adherence by considering the drug, the dose, and the regimen separately produced quantifiable data concerning the subjects' medication habits. Non-adherence ranged from 20 to 70% depending on the measuring method. The participants' knowledge of the treatment was poor. Our results suggest that better information on medication and the use of compliance aids may prevent nonadherence. Special attention should be paid to persons receiving three or more drugs, living alone, receiving drugs from other doctors, and to persons with predementia symptoms, as they are at higher risk of nonadherence. PMID- 11422023 TI - Recording previous adverse drug reactions--a gap in the system. AB - AIMS: To measure the accuracy of recording of previous adverse drug reaction (ADR) history in patients admitted to a teaching hospital before and after an education programme. METHODS: One month survey of patients on one medical and one surgical ward, repeated after a 1 month education programme. Patients answered a questionnaire about previous ADRs and this information was compared with that in all relevant sections of their medical records and medication charts. RESULTS: Of 117 patients at baseline, 50 had a total of 81 previous ADRs. Only 75% were recorded on medication charts and 57% and 64%, respectively, in medical and nursing notes. In the post education survey of 124 patients, 56 had 105 previous ADRs, 85% were recorded on medication charts and 64% and 70% in medical and nursing records. These differences were not significant. Serious ADRs were also poorly recorded at baseline but, due to intervention by ward pharmacists, their recording on medication charts improved significantly after education. Pharmacists also significantly improved the quality of description of previous ADRs in both parts of the study. CONCLUSIONS: Previous ADR history obtainable from hospital patients is poorly recorded in medical records and an intensive education programme only produced a significant change in recording by ward pharmacists. Better strategies are needed to improve this essential aspect of history taking. PMID- 11422024 TI - Is cytochrome P450 2C9 genotype associated with NSAID gastric ulceration? AB - AIMS: The aim of this study was to explore whether genetic variation of cytochrome P450 2C9 (CYP2C9) contributes to NSAID-associated gastric ulceration. The hypothesis tested was that CYP2C9 poor metabolizer genotype would predict higher risk of gastric ulceration in patients on NSAIDs that are metabolized by CYP2C9, due to higher plasma NSAID concentrations. METHODS: Peripheral blood DNA samples from 23 people with a history of gastric ulceration attributed to NSAIDs metabolized by CYP2C9, and from 32 people on NSAIDs without gastropathy, were analysed to determine CYP2C9 genotype. RESULTS: The following genotypes were found: *1/*1 (wild type) in 70% of cases and 58% of controls, *1/*2 in 17% of cases and 29% of controls, *1/*3 in 13% of cases and 13% of controls. The difference between case and control nonwild-type genotype frequency was 11.5% (95% CI -14,37%), with the direction of the difference being against the hypothesis. No individuals with homozygote poor metaboliser genotype were identified. The differences in genotype frequencies between the two groups were not significant and the frequencies were similar to those in a large published population study. Ninety-five percent binomial confidence interval analysis confirms that there is no apparent clinically significant relationship between CYP2C9 genotype and risk of gastric ulceration although a small difference in risk in poor metabolizers cannot be excluded. CONCLUSIONS: These results do not support the hypothesis that gastric ulceration resulting from NSAID usage is linked to the poor metabolizing genotypes of CYP2C9. PMID- 11422025 TI - The effect of fluconazole on ritonavir and saquinavir pharmacokinetics in HIV-1 infected individuals. AB - AIMS: To study the effect of fluconazole on the steady-state pharmacokinetics of the protease inhibitors ritonavir and saquinavir in HIV-1-infected patients. METHODS: Five subjects treated with saquinavir and three with ritonavir received the protease inhibitor alone (saquinavir 1200 mg three times daily, ritonavir 600 mg twice daily) on day 1, and the same protease inhibitor in combination with fluconazole (400 mg on day 2 and 200 mg on days 3 to 8). Pharmacokinetic parameters were determined on days 1 and 8. RESULTS: In the saquinavir group, the median increase in the area under the plasma concentration vs time curve was 50% from 1800 microg l(-1) h to 2700 microg l(-1) h (P = 0.04, median increase: 900 microg l(-1) h; 2.5 and 97.5 percentile: 500-1300), and 56% for the peak concentration in plasma (from 550 to 870 microg l(-1), P = 0.04; median increase: 320 microg l(-1) h, 2.5 and 97.5 percentile: 60-450 microg l(-1)). In the ritonavir group, there were no detectable changes in the pharmacokinetic parameters on addition of fluconazole. CONCLUSIONS: Because of the favourable safety profile of saquinavir, dose adjustments are probably not necessary with concomitant use of fluconazole, as is the case for ritonavir. PMID- 11422026 TI - Are we going OTT about ITT? PMID- 11422027 TI - Paraneoplastic pemphigus: the role of tumours and drugs. PMID- 11422028 TI - Cellular localization of fractalkine at sites of inflammation: antigen-presenting cells in psoriasis express high levels of fractalkine. AB - BACKGROUND: Chemokines play a key role in cell trafficking at sites of inflammation. The fractalkine CX3C chemokine is unique in several aspects. Fractalkine is expressed on activated endothelial cells and exists in two forms, either membrane anchored or in a soluble form. The soluble form is a potent chemotactic agent for T cells/monocytes and the anchored form functions as an adhesion molecule. In view of these specific functions fractalkine is capable of controlling the key regulatory mechanisms of cell trafficking at sites of inflammation. OBJECTIVES: Little is known about the significance of this important molecule in inflammatory diseases. We undertook this study to elucidate the role of fractalkine in inflammatory diseases of the skin. METHODS: We used a polyclonal antifractalkine antibody (immunoperoxidase and immunofluorescence stainings) in cryosections obtained from tissues of normal skin and that of selected cutaneous inflammatory diseases (psoriasis, lichen planus, eczema). RESULTS: Increased expression of fractalkine was observed in the dermal blood vessels of lichen planus, eczema and psoriasis tissues. The most striking finding was that the dermal dendrocytes in the papillary dermis of psoriasis tissues expressed high levels of fractalkine. Compared with 186.64 +/- 51.69 fractalkine positive dermal dendrocytes per mm2 of the upper dermis of psoriatic tissue, the number of positive cells in lichen planus, eczema, and normal skin were 17.29 +/- 12.50, 12.50 +/- 6.75 and 5.93 +/- 3.53, respectively. We also performed double label immunofluorescence staining with nerve growth factor receptor (NGF-R) antibody and fractalkine antibody. NGF-R-positive terminal cutaneous nerves were in close contact with the fractalkine-positive dermal dendrocytes in psoriatic lesions. CONCLUSIONS: The results of this study confirm that fractalkine is upregulated at sites of inflammation. Thus, it is likely that this molecule plays a key part in cell trafficking. An increased expression of fractalkine at the dermal papillae provides a plausible explanation for the migration and accumulation of T cells at these sites in psoriasis. Earlier studies have reported an increased number of dermal dendrocytes in psoriatic tissue; however, the functional role of these cells in the pathogenesis of psoriasis is largely unknown. Expression of fractalkine on the surface of dermal dendrocytes suggests an active role for these cells in localization and activation of lesional T cells. PMID- 11422029 TI - The antipsoriatic drug dimethylfumarate strongly suppresses chemokine production in human keratinocytes and peripheral blood mononuclear cells. AB - BACKGROUND: The effectiveness of systemic treatment of psoriasis with fumaric acid esters has been proven, but their mode of action at the cellular and molecular level has not yet been fully elucidated. OBJECTIVES: To study the effect of dimethylfumarate (DMF) on the production of the chemokines CXCL1, CXCL8, CXCL9, CXCL10 and CXCL11, formerly known as GROalpha, interleukin-8, Mig, IP-10 and IP-9/I-TAC, respectively, in human keratinocytes and peripheral blood mononuclear cells (PBMC). METHODS: Cultured keratinocytes were stimulated with interferon (IFN) -gamma to produce CXCL9, CXCL10 and CXCL11 and with phorbol myristate acetate to produce CXCL1 and CXCL8 in the absence and presence of DMF (5, 15 and 45 micromol L(-1)). PBMC were stimulated with either IFN-gamma to produce CXCL9 and CXCL10 or lipopolysaccharide to produce CXCL8, in the absence and presence of DMF (5, 15 and 45 micromol L(-1)). RNA preparations from isolated keratinocytes were analysed by Northern blotting; protein production by keratinocytes and PBMC was monitored by an enzyme-linked immunosorbent assay. RESULTS: Northern blot analysis on isolated keratinocyte RNA preparations showed a dose-dependent inhibition of CXCL1, CXCL8, CXCL9, CXCL10 and CXCL11 transcription by DMF. At 45 micromol L(-1) the inhibition was almost complete. In addition, keratinocytes and PBMC showed in the presence of DMF a dose-dependent inhibition of CXCL8, CXCL9 and CXCL10 protein production. CONCLUSIONS: These results show the ability of DMF to inhibit the production of chemokines that may be critically involved in the development and perpetuation of psoriatic lesions. This might explain, at least in part, the beneficial effects of treatment with fumaric acid esters in psoriasis patients. PMID- 11422030 TI - Effect of propylthiouracil on adenosine deaminase activity and thyroid function in patients with psoriasis. AB - BACKGROUND: T-cell activation has been implicated in the pathogenesis of psoriasis; adenosine deaminase (ADA) activity has been considered as a marker of T-cell activation. The antithyroid drug propylthiouracil (PTU) has recently been shown to have beneficial effects on psoriatic lesions, probably by acting on the immune system. OBJECTIVES: To investigate whether ADA activity may be related to psoriasis and whether oral PTU affects ADA activity and gives clinical improvement in psoriatic patients. METHODS: ADA activities were measured in plasma, erythrocyte and tissue samples of patients with psoriasis before and after 2 months of treatment with either PTU 100 mg three times daily or PTU plus thyroxine 25 microg once daily (to prevent possible hypothyroidism, which may be induced by PTU) as well as in healthy controls. The severity of the disease was evaluated before and after treatment according to Psoriasis Area and Severity Index (PASI) scores. Routine analyses and thyroid function tests were also carried out during the study. RESULTS: All patients showed significant clinical improvement in their lesions and decreased PASI scores after the treatments. Elevated baseline ADA activities in skin and plasma were found to be lower, and decreased baseline erythrocyte ADA was higher, after the treatments in all patients, and they were not different from control values. Although thyroid function tests were not affected by the treatments, serum thyroid-stimulating hormone levels were found to be higher after the treatments, and there was a larger increase in patients treated with PTU alone. However, none of the patients had clinical hypothyroidism or cytopenia. CONCLUSIONS: ADA activity may be clinically useful for indicating T-cell activation in psoriasis. Because of its antiproliferative and immunomodulatory effects, antioxidant potential and low toxicity, PTU may be an effective agent in the treatment of psoriasis. PMID- 11422031 TI - Liver cytochrome P450 CYP1A2 is markedly inhibited by systemic but not by bath PUVA in dermatological patients. AB - BACKGROUND: Methoxsalen (8-MOP) may cause important pharmacokinetic drug interactions as it has been shown to inhibit and/or induce several drug metabolizing enzymes in vitro, in animal models and in humans. OBJECTIVES: In order to assess the clinical importance of acute and chronic 8-MOP effects on the liver cytochrome P-450 enzyme CYP1A2 in vivo, we measured caffeine clearance in dermatological patients before the onset of systemic or bath psoralen + ultraviolet A radiation (PUVA) (8-MOP + UVA) therapy, on the first day and after 1 week of treatment. METHODS: Data from four patients with systemic PUVA and seven patients with bath PUVA were available (age range 23-71 years, five women and six men). RESULTS: For all of the patients, individual pre-PUVA caffeine clearance values were above the lower limit of previously assessed reference ranges. Systemic PUVA markedly decreased caffeine clearance by factors of 0.17 [90% confidence interval (CI) 0.07-0.42] on the first day and 0.14 (90% CI 0.05 0.36) after 1 week of treatment, respectively, and values thus dropped below the reference ranges. In contrast, bath PUVA had no obvious effect on pre-PUVA clearance values as the latter changed by factors of 1.00 (90% CI 0.81-1.23) and 1.05 (90% CI 0.75-1.49) on the first day and after 1 week of treatment, respectively. CONCLUSIONS: Systemic PUVA causes pronounced inhibition of liver CYP1A2, while bath PUVA has no such effect. The extent of interaction makes a dose adjustment for most CYP1A2 substrates such as theophylline mandatory in patients undergoing systemic PUVA. PMID- 11422032 TI - Perforin and granzyme B may contribute to skin inflammation in atopic dermatitis and psoriasis. AB - BACKGROUND: Infiltration of the skin by pathogenic T cells is regarded as a key factor in the development of inflammatory skin diseases such as atopic dermatitis (AD) and psoriasis. OBJECTIVES: To investigate whether T cells containing cytotoxic proteins may contribute to the generation of skin inflammation in these skin diseases. METHODS: Skin biopsy specimens were obtained from non-lesional and lesional skin of patients with chronic AD (n = 8) and psoriasis (n = 6), and from non-atopic controls with normal skin (n = 6). Expression of perforin and granzyme B was investigated by immunohistochemistry. RESULTS: A significant enhancement of perforin and granzyme B expression was observed in lesional AD skin as compared with normal skin, non-lesional AD skin and psoriasis. Expression of these cytotoxic proteins was also increased in psoriasis as compared with normal skin and non-lesional psoriatic skin. Immunoreactivity for perforin and granzyme B was mainly found in the cytoplasm of lymphocytic cells located in the perivascular infiltrate. In AD increased numbers of positive cells were also observed focally at sites of spongiosis in the epidermis. Double immunostaining revealed that both CD4+ and CD8+ T cells are capable of expressing perforin and granzyme B. CONCLUSIONS: Our data suggest that cytotoxic CD4+ and CD8+ T cells containing perforin and granzyme B may play an integral part in eliciting cutaneous inflammation in AD. PMID- 11422033 TI - CD4+ CD8+ (thymocyte-like) T lymphocytes present in blood and skin from patients with atopic dermatitis suggest immune dysregulation. AB - BACKGROUND: Atopic dermatitis (AD) is a chronic inflammatory skin disease expressed early in life. Disease development is primarily determined by as yet unknown genetic factors, leading to the accumulation of activated T lymphocytes in the skin. OBJECTIVES: To investigate the nature of these T cells. METHODS: T cell lines could be established from AD skin biopsies, but not from normal skin or AD peripheral blood, when placed in RPMI 1640 medium with 10% human AB serum, antibiotics, and the T-lymphocyte growth factors interleukins 2 and 4. The cell lines were subjected to phenotypic analysis using a fluorescence-activated cell sorter and compared with lymphocytes from AD and normal control peripheral blood. RESULTS: T-cell lines from 22 of 24 consecutive skin biopsies taken from 24 adult patients with AD were established. All cells were T lymphocytes expressing several activation markers. A significant proportion of the lymphocytes had stable expression of a CD4+ CD8+ phenotype (26% +/- 6%; mean +/- SEM). Such double-positive T lymphocytes are normally only seen in the thymus and not in the peripheral immune system. CD4+ CD8+ cells in peripheral blood of the patients (12.5% +/- 3.3%) were also detected. CONCLUSIONS: We suggest that a basic pathophysiological change in AD may be a faulty maturation of the T-lymphocyte system, leading to skin inflammation with CD4+ CD8+ T lymphocytes resembling immature T cells. This is likely to lead to skewing of many immune reactions in the patients. PMID- 11422034 TI - Serum levels of soluble stem cell factor and soluble KIT are elevated in patients with atopic dermatitis and correlate with the disease severity. AB - BACKGROUND: Mast cell infiltration in skin lesions of atopic dermatitis (AD) is considered to play an important role in the pathogenesis of the disease. The most common factor that stimulates mast cell growth, migration and differentiation is stem cell factor (SCF), and the interaction of SCF and its receptor, KIT (tyrosine kinase transmembrane receptor), appears to be the key event in the recruitment and proliferation of mast cells. OBJECTIVES: To determine whether any altered metabolism of SCF and/or KIT is present in patients with AD. METHODS: We measured serum levels of soluble SCF (sSCF) and soluble KIT (sKIT) using enzyme linked immunosorbent assay in 54 patients with AD, five patients with erythrodermic psoriasis vulgaris and 64 healthy individuals. RESULTS: Serum levels of both peptides in AD patients were significantly higher than those in healthy individuals, whereas patients with psoriasis vulgaris did not show any difference from healthy controls. Both sSCF and sKIT levels were positively correlated with the disease severity in AD patients, and decreased after effective treatment with topical corticosteroids. Conclusion Serum levels of sSCF and sKIT may be useful indicators for evaluation of the activity and severity of AD. PMID- 11422035 TI - Differences in efficacy between intention-to-treat and per-protocol analyses for patients with psoriasis vulgaris and atopic dermatitis: clinical and pharmacoeconomic implications. AB - BACKGROUND: Pharmacoeconomic outcome research is based on three criteria: (i) evaluation of objective therapeutic effects; (ii) quality of life; and (iii) treatment costs. Evaluation of therapeutic effect is mainly based on the results of clinical trials using objective clinical measures, e.g: Psoriasis Area and Severity Index (PASI) (score for psoriasis vulgaris) and the Severity Scoring of Atopic Dermatitis (SCORAD) (score for atopic dermatitis). In most studies, only results for a treatment-optimized subpopulation (patients treated according to the protocol) are presented in publications. The relevance of such data for daily routine therapy is doubtful. OBJECTIVES: Our purpose was to investigate the expected loss of effectiveness of switching from a clinical trial to daily routine therapy for the synchronous application of narrow-band ultraviolet (UV) B phototherapy (311 nm) and bathing in 10% Dead Sea salt solution (synchronous balneophototherapy) for patients with psoriasis vulgaris and atopic dermatitis. METHODS: We conducted a multicentre, uncontrolled observational study of outpatients. To achieve data for 'clinical trial' and 'daily routine' situations, two populations were compared: (i) all patients strictly treated according to the protocol (ATP) with no protocol deviations (data published in clinical trials), and (ii) all patients participating in the study who received active treatment at least once, despite treatment irregularities, non-compliance, early withdrawal or other protocol violations [intention-to-treat-population (ITT), model for 'daily routine']. RESULTS: A total of 2526 patients were included in the ITT analysis for psoriasis vulgaris (n = 487 for atopic dermatitis), of which 818 patients could be analysed according to protocol (n = 104 for atopic dermatitis). Striking differences in the therapeutic effect between both groups (ITT and ATP) were found using relative PASI and SCORAD score improvement: 11% (57% 'daily routine' vs. 68% in 'clinical trial') for psoriasis vulgaris and 16% (39% 'daily routine' vs. 55% 'clinical trial') for atopic dermatitis. The main reasons for excluding patients from the 'clinical trial' group were early study withdrawal in 29% (atopic dermatitis, 47%) of patients and fewer treatments per week than planned in the protocol in 24% (atopic dermatitis, 52%). CONCLUSIONS: Our data clearly indicate that for the prediction of the therapeutic effect for daily routine therapy the ITT data appear to be more relevant than the ATP results (i.e. those presented in clinical trials). Although these data are only a first step for evaluating the 'real' therapeutic effect of a treatment modality in daily routine, they seem to support the requirements for ITT analyses in efficacy studies and demonstrate the necessity of ITT data for pharmacoeconomic evaluation. PMID- 11422036 TI - Ultraviolet AI exposure of human skin results in Langerhans cell depletion and reduction of epidermal antigen-presenting cell function: partial protection by a broad-spectrum sunscreen. AB - BACKGROUND: Ultraviolet (UV) B-induced effects on the skin immune system have been extensively investigated, but little is known regarding the immunological changes induced by UVA exposure of human skin. Recent data assessing the protection afforded by sunscreens against photoimmunosuppression stress the need for broad-spectrum sunscreens with an adequate UVA protection. OBJECTIVES: The purpose of this study was first to determine the changes observed in epidermal Langerhans cells (ELC) density and epidermal antigen-presenting cell (APC) activity after exposure of human skin to UVAI (340-400 nm) radiation, and secondly to assess the immune protection afforded in vivo by a sunscreen formulation containing a long wavelength UVA filter with a low UVA protection factor (UVA-PF = 3). METHODS: Epidermal cell (EC) suspensions were prepared from skin biopsies 3 days after exposure to a single dose of UVAI (either 30 or 60 J cm(-2)). RESULTS: Flow-cytometric analysis of EC suspensions revealed that exposure to 60 J cm(-2) UVAI resulted in a decreased number of ELC without infiltration of CD36+ DR+ CD1a- antigen-presenting macrophages into the epidermis, and a significant reduction of HLA-DR expression on viable ELC. In vivo exposure to both 30 and 60 J cm(-2) resulted in a decreased allogeneic CD4+ T-cell proliferation induced by UVAI-irradiated ECs. The sunscreen application partially prevented (57 +/- 9%) the decrease in epidermal allogeneic APC activity induced by 60 J cm(-2) UVAI. CONCLUSIONS: In vivo UVAI exposure of human skin results in a decreased number of ELC and in a downregulation of epidermal APC activity. This last effect is partially prevented by prior application of a sunscreen with a low UVAI-PF value. These results indicate that increasing the absorption of UV filters for long UVA wavelengths may lead to an improved immune protection. PMID- 11422037 TI - Cutaneous side-effects in cancer patients treated with the antiepidermal growth factor receptor antibody C225. AB - BACKGROUND: C225 is an antibody to the epidermal growth factor receptor (EGFR), and inhibits growth of various tumour cells. The antibody is currently being used as a therapeutic agent in several clinical trials of patients with carcinomas. Objectives To determine and investigate the cutaneous side-effects in cancer patients treated with C225. Methods We clinically examined 10 patients treated with C225, and performed immunohistochemical and in situ hybridization studies on skin biopsies. Results The most common cutaneous reaction to C225 therapy was the development of an acneiform follicular eruption, which was most pronounced on the face, chest and upper back and typically manifested a week after the onset of treatment. The consistency of the morphology and timing of the clinical findings in 10 different patients following monotherapy with C225 strongly suggested a direct biological effect of the antibody. Additional dermatological side-effects included focal areas of tender paronychial inflammation of toes and fingers and small aphthous ulcers of the oral mucosa. Serial punch biopsies of chest skin before and after treatment (at 8 days) revealed two main reaction patterns: a superficial dermal inflammatory cell infiltrate surrounding hyperkeratotic and ectatic follicular infundibula, and a suppurative superficial folliculitis. In two biopsies focal intraepidermal acantholysis was found. Microbiological cultures failed to reveal an infectious aetiology. Immunohistochemical and in situ hybridization studies on a subset of the biopsies showed an increase in the expression of p27Kip1 in epidermal keratinocytes after treatment with C225. Conclusions Our findings support the concept that p27Kip1 plays a part in the in vivo regulation of follicular and epidermal homeostasis by EGFR. PMID- 11422038 TI - The clinical transition between pemphigus foliaceus and pemphigus vulgaris correlates well with the changes in autoantibody profile assessed by an enzyme linked immunosorbent assay. AB - BACKGROUND: There are a number of reports of pemphigus with clinical shifting between pemphigus foliaceus (PF) and pemphigus vulgaris (PV). On the other hand, a novel enzyme-linked immunosorbent assay (ELISA) against recombinant baculoproteins of desmoglein 1 (Dsg1) (PF antigen) and Dsg3 (PV antigen) has been established and found to be extremely sensitive and specific. OBJECTIVES: To characterize the change in the antibody profiles in a series of pemphigus cases with mixed features of PF and PV by various methods, including the novel ELISA. Patients/methods Sera were obtained from eight cases undergoing a shift between PF and PV and three cases of coexistent PF and PV. The autoantigens were analysed by ELISA, as well as by immunofluorescence using normal human skin sections and immunoblotting using normal human epidermal extracts. RESULTS: The results of the ELISA, immunofluorescence and immunoblotting studies showed that the transition between PF and PV correlates well with the changes of autoantibodies against either Dsg1 or Dsg3. CONCLUSIONS: The clinical phenotype at each stage is defined by the anti-Dsg antibody profile in the serum of these pemphigus patients showing mixed features of PF and PV. In addition, ELISA using recombinant baculoproteins was particularly useful in distinguishing PF and PV. PMID- 11422039 TI - IgG, IgA and IgE autoantibodies against the ectodomain of desmoglein 3 in active pemphigus vulgaris. AB - BACKGROUND: IgG autoantibodies against desmoglein (Dsg) 3 play a key part in the pathogenesis of pemphigus vulgaris (PV), the most severe autoimmune bullous disorder. OBJECTIVES: To determine whether immunoglobulin isotypes other than IgG are detectable in the sera of patients with PV and whether a particular immunoglobulin subtype is associated with a distinct clinical phenotype of PV. METHODS: Sera from 41 patients with acute-onset, chronic active, and remittent PV disease with mucosal and cutaneous lesions were assayed against a baculovirus expressed Dsg3 protein by immunoblot analysis. RESULTS: In acute-onset PV, Dsg3 reactive IgG1 was detected in nine of 15 (60%), IgG4 in 14 of 15 (93%), IgA in nine of 15 (60%) and IgE in two of 15 (13%) sera. In chronic active PV, Dsg3 reactive IgG1 was detected in 11 of 18 (61%), IgG4 in 16 of 18 (89%), IgA in 13 of 18 (72%) and IgE in two of 18 (11%) sera. In contrast, sera from patients with remittent PV disease contained only Dsg3-reactive IgG1 in six of eight (75%) and IgG4 in four of eight (50%) cases, but not Dsg3-reactive IgA or IgE. CONCLUSIONS: In extension of previous findings, our study demonstrates that, in addition to IgG autoantibodies, IgA and occasionally IgE autoantibodies reactive with Dsg3 are present in acute and chronic active PV. The detection of Dsg3-reactive autoantibodies of the IgG4, IgA and IgE subclasses in active PV provides additional evidence that PV is a T-helper 2-regulated autoimmune disorder. PMID- 11422040 TI - Immunohistochemical study of angiotensin receptors in normal human sweat glands and eccrine poroma. AB - BACKGROUND: Angiotensin II exerts its actions through its specific receptors. However, expression of these receptors has not been determined in sweat glands. OBJECTIVES: To clarify the expression and localization of the angiotensin receptors in normal human sweat glands and eccrine poroma. METHODS: Expression of angiotensin type 1 (AT1) and type 2 (AT2) receptors in normal human eccrine and apocrine sweat glands and 12 cases of eccrine poroma was studied using immunohistochemistry. RESULTS: In eccrine sweat glands, the acrosyringium and the inner surfaces and luminal cells of the intradermal duct showed positive staining with AT1. In apocrine sweat glands, the intraepithelial duct and luminal cells of the intradermal duct showed positive staining with AT1. In 12 cases of eccrine poroma, some of the tumour cells in the tumour strands and cells surrounding the luminal structures stained positively. There were no positive findings with AT2. CONCLUSIONS: Studying AT1 distribution may be useful in understanding the pathophysiology of sweat glands and sweat gland tumours. PMID- 11422041 TI - Pattern of expression of c-Myc, Max and Bin1 in human anagen hair follicles. AB - BACKGROUND: We recently reported the presence of c-Myc immunoreactivity in two distinct regions of the inner root sheath (IRS) of human anagen hair follicles; they corresponded to the regions where keratinocytes of Henle's and Huxley's layers enter the terminal differentiation phase that will lead to their exfoliation in the pilary canal. These regions were denoted lower (LR) ring and upper ring (UR). OBJECTIVES: To extend these observations to other genes connected to c-Myc and specifically to Max and Bin1. Max is the best known heterodimeric partner of c-Myc, interacting with its C-terminal domain, and Bin1 is an adaptor protein interacting with its N-terminal domain. METHODS: Human anagen hair follicles were processed for c-Myc, Max and Bin1 immunohistochemistry and immunofluorescence. The presence of different isoforms of Bin1 was evaluated by Western blot analysis. RESULTS: Analysis of sections cut in several planes, including tangential, demonstrated the presence of a third ring of c-Myc-positive cells (intermediate ring; IR) in the cuticle of the IRS corresponding to the region where this thin layer undergoes keratinization. Max immunoreactivity was observed in the three layers of the IRS starting in the lower bulbar region and ending in each of them at the level of the corresponding c-Myc-positive ring. Bin1 immunoreactivity was clearly distinguished only in Huxley's layer and in the cuticle, starting in some cells below the UR and terminating at the level of the latter. The companion layer of the outer root sheath was also labelled up to the infundibular region. Max and Bin1 immunostaining were less consistently observed in other skin adnexae and in the epidermis. CONCLUSIONS: The results indicate that the asynchronous differentiation along the axis of the hair follicle of the different layers of the IRS and of the companion layer involves the expression of different genes that are interrelated in the so-called 'Myc network'. The specific localization of c-Myc in the IRS only at the level of the discrete and limited regions of the three rings appears to be the hallmark of the switch from differentiation to terminal differentiation/cell deletion. PMID- 11422042 TI - Genital squamous cell carcinoma in men treated by photochemotherapy. A cancer registry-based study from 1978 to 1998. AB - BACKGROUND: One single report from the U.S. 16-centre-trial indicated that psoralen and ultraviolet A radiation (PUVA) therapy may induce an increased risk of genital tumours in men, and protection of the genital area is, therefore, recommended. OBJECTIVES: To evaluate the relevance of this risk in routine clinical practice. METHODS: Two groups of patients were included in a 1978-98 retrospective study. Case records of men with genital squamous cell carcinoma (SCC) identified from the Cancer Registry of the Doubs area of France were examined for a history of PUVA therapy, topical tar treatment, psoriasis, human papillomavirus infection or genital dermatitis. In addition, all the dermatologists of the Doubs area (in public and private practice) using PUVA therapy were asked to provide information on the number of patients having received PUVA therapy and whether the genital area was exposed during treatment. RESULTS: Between 1978 and 1998, among the 48 men who had developed a genital SCC in the Doubs area, only one had a history of intensive PUVA therapy. About 150,000 treatments with PUVA therapy had been performed by 15 dermatologists in the Doubs area for 5400 patients since 1978. No case of genital SCC had been reported, despite the fact that the genital area had not been protected during UVA exposure. CONCLUSIONS: Although retrospective, our study demonstrates that the occurrence of genital SCC in men treated with PUVA therapy is a very rare event in common dermatological practice. PMID- 11422043 TI - Systemic photodynamic therapy with aminolaevulinic acid delays the appearance of ultraviolet-induced skin tumours in mice. AB - BACKGROUND: Photodynamic therapy (PDT) with topical aminolaevulinic acid (ALA) has recently been approved by the US Food and Drug Administration for the treatment of actinic keratoses. OBJECTIVES: To determine whether weekly systemic suberythemogenic ALA-PDT could prevent the appearance of ultraviolet (UV) induced skin tumours in hairless mice. METHODS: One group of 20 mice received daily UV radiation from FS 20 tubes, and weekly intraperitoneal injections of ALA 40 mg kg(-1), each followed 3 h later by 12 J cm(-2) of white light (ALA-PDT). Control groups consisted of mice exposed only to UV, to UV and ALA without white light, or UV and white light without ALA, as well as untreated mice. RESULTS: The tumour-free survival was significantly longer for mice exposed to daily UV and weekly ALA-PDT as compared with the control groups. Neither the mortality nor the incidence of large skin tumours was higher in the UV/ALA-PDT group than in mice exposed only to UV. In vivo fluorescence spectroscopy showed that the 635-nm fluorescence emission within tumours was lower than in normal skin 3 h after ALA administration. This was also confirmed by quantitative fluorescence microscopy. CONCLUSIONS: Systemic ALA-PDT can delay the appearance of UV-induced skin tumours in mice without increasing mortality or the incidence of large tumours. PMID- 11422044 TI - A case of Rombo syndrome. AB - Rombo syndrome is a rare entity characterized by the presence of atrophoderma vermiculatum of the face, multiple milia, telangiectases, acral erythema and a propensity to develop basal cell carcinomas. We describe a patient whose clinical and histopathological abnormalities are consistent with this diagnosis. PMID- 11422045 TI - Systemic lupus erythematosus presenting with oral mucosal lesions: easily missed? AB - Two caucasian patients are described in whom oral mucosal lesions were the first manifestation of systemic lupus erythematosus. In both cases the diagnosis was delayed despite histological examination of oral lesions. Treatment with antimalarials and azathioprine was of significant benefit. In the absence of cutaneous or systemic features, distinguishing oral lupus erythematosus from lichen planus and epidermal dysplasia can be difficult, both clinically and on histology, and requires a high index of suspicion. PMID- 11422046 TI - Pyoderma vegetans and ulcerative colitis. AB - Pyoderma vegetans (PV) is a chronic, vegetating pustular disorder characterized clinically by erythematous vesiculopustular vegetating cutaneous plaques. Marked epidermal hyperplasia, intraepidermal and subepidermal neutrophilic microabscesses and a dermal inflammatory infiltrate are the prominent histopathological findings. We describe a patient with PV associated with ulcerative colitis and mammary Paget's disease. Pustular eruptions associated with ulcerative colitis are reviewed. PMID- 11422047 TI - Chronic urticaria associated with intra-articular methylprednisolone. AB - Reports of allergic reactions following treatment with systemic corticosteroids are rare, despite their widespread use. A 47-year-old man developed widespread urticaria, resistant to antihistamines, coinciding with injections of local anaesthetic and methylprednisolone for cervical spondylosis. He underwent immediate and delayed hypersensitivity tests. Intradermal tests showed immediate type sensitivity to methylprednisolone and hydrocortisone. Patch tests were positive to 21 of 26 corticosteroids tested. A diagnosis of both an immediate and a delayed-type hypersensitivity to corticosteroids was made. With avoidance of all corticosteroids he has been free from urticarial rash for 9 months and has been able to stop all medication. PMID- 11422048 TI - Anticonvulsant hypersensitivity syndrome associated with reactivation of cytomegalovirus. AB - Recently, it has been proposed that viral infection is involved in the pathogenesis of hypersensitivity syndrome. Cytomegalovirus (CMV), one of the aetiological agents of infectious mononucleosis, has never been reported as an organism associated with hypersensitivity syndrome. We describe a 64-year-old man with severe phenytoin-induced hypersensitivity syndrome associated with CMV infection. Twenty-five days after the patient was started on phenytoin, he developed high fever and a generalized erythematous rash followed by jaundice, renal failure and disseminated intravascular coagulopathy (DIC). CMV-specific IgG antibodies were significantly increased 7 weeks after the onset of clinical symptoms and the increase was associated with the appearance of CMV-specific IgM. CMV DNA was detected in the serum of the patient. Coinfection with other viruses, such as Epstein-Barr virus and human herpesviruses 6 and 7, could be excluded because antibody titres to those viruses did not increase during the clinical course of his illness. We suggest that reactivation of CMV may contribute, at least in some cases, to the development of hypersensitivity syndrome. PMID- 11422049 TI - Sodium valproate-induced cutaneous pseudolymphoma followed by recurrence with carbamazepine. AB - We report a patient with a sodium valproate-induced cutaneous pseudolymphoma, presenting with an erythematous papule, histologically mimicking a non epidermotropic T-cell lymphoma. Polymerase chain reaction study of the skin biopsy revealed monoclonal rearrangement of the T-cell receptor gamma gene. Withdrawal of sodium valproate was followed by regression of the lesion, but 5 months after substitution by carbamazepine, two further papules appeared, with similar histological features and a T-cell clone identical to the initial one. Carbamazepine was stopped and the lesions disappeared without relapse over a 4 year follow-up. Sodium valproate is very rarely responsible for a hypersensitivity syndrome, and our case is the first report of sodium valproate induced cutaneous pseudolymphoma. The recurrence with carbamazepine may be due to a common effect on T-cell lymphocyte function. The return of the same monoclonal population shows that the recurrence of monoclonal T cells may be observed in benign conditions and is not an exclusive hallmark of cutaneous lymphoma. PMID- 11422050 TI - Intralesional rituximab for cutaneous B-cell lymphoma. AB - Rituximab, a chimeric anti-CD20 monoclonal antibody, has been approved for systemic treatment of relapsed or refractory CD20-positive B-cell non-Hodgkin's lymphoma. As cutaneous B-cell lymphoma (CBCL) also expresses the CD20 molecule, three patients with histologically and immunohistochemically confirmed CBCL without systemic involvement were treated with low-dose intralesional rituximab in a pilot study. Single doses applied ranged from 10 to 30 mg per lesion, according to lesion extent, with a cumulative dose of up to 350 mg. Injections were given two or three times weekly for 3-5 weeks, with a second cycle after 6 weeks in one patient with incomplete remission. Complete and lasting remission was achieved in each patient; this has persisted for up to more than 1 year. The observed adverse events were of grade 1 severity. Results suggest that intralesional rituximab may be a safe and effective new therapy modality for CBCL. PMID- 11422051 TI - Lymphoma-type adult T-cell leukaemia-lymphoma with a bulky cutaneous tumour showing multiple human T-lymphotropic virus-1 DNA integration. AB - Human T-lymphotropic virus-1 (HTLV-1) is considered to be the cause of adult T cell leukaemia-lymphoma (ATL). Monoclonal integration of HTLV-1 proviral DNA, as is analysed by Southern blotting, has been demonstrated in ATL patients. Unusual integration patterns of HTLV-1 proviral DNA have occasionally been described, and it is suggested that the patterns have clinical implications for ATL pathophysiology. Multiple, complete and defective types of integration patterns, in that order, are apparently associated with prognoses from good to poor. We report a 73-year-old Japanese woman with lymphoma-type ATL and a bulky cutaneous tumour on the left thigh. Four bands of slightly differing intensity were seen after EcoRI digestion of skin and lymph node samples on Southern blot analysis of HTLV-1 proviral DNA. Analysis for T-cell receptor-beta gene revealed five novel bands after restriction enzyme digestion with HindIII, indicating that the patient has four separate tumour cell clones, each of which carries one copy of the provirus. She was treated with chemotherapy and radiation and remains under reasonable control despite some relapsing cutaneous nodules. The indolent course in this present case could be related to the multiple integration pattern of HTLV 1 proviral DNA detected. PMID- 11422052 TI - A localized variant of paraneoplastic pemphigus: acantholysis associated with malignant melanoma. AB - We report a 72-year-old male patient with a nodular malignant melanoma that was associated with focal suprabasal acantholysis (FSA). This phenomenon, which is regarded as an incidental finding by dermatopathologists, may be associated with inflammatory and also neoplastic skin diseases. Haematoxylin and eosin stained sections from an erythematous plaque surrounding the patient's tumour showed FSA, direct immunofluorescence (DIF) and indirect immunofluorescence (IIF) on normal human skin, monkey oesophagus and rat urinary bladder were negative. On electron microscopy few desmosomes could be detected in the basal cell layer of the acantholytic areas and there was a nearly complete loss of these structures in the spinous cell layer. Only remnants of cytoplasmic plaques and keratin filaments could be observed in those areas. In contrast, adherens junctions appeared to be well preserved. An enzyme-linked immunosorbent assay (ELISA) using recombinant fusion proteins as antigens did not show circulating autoantibodies against desmoglein 1 (Dsg1) or desmoglein 3 (Dsg3). In contrast, immunoblotting revealed autoantibodies directed against keratinocyte antigens with a molecular weight of 85 kDa and 250 kDa, the first band corresponding to the molecular weight of comigrating plakoglobin. Immunoprecipitation with patient serum also revealed a 85-kDa band. We conclude that these autoantibodies, probably in conjunction with cofactors produced by the tumour, could play a part in the pathogenesis of this variant of FSA, for which we propose the term 'localized paraneoplastic pemphigus.' PMID- 11422053 TI - Paraneoplastic pemphigus: an association with fludarabine? AB - Paraneoplastic pemphigus is a relatively recently described immunobullous disease with characteristic features. We report three cases of paraneoplastic pemphigus in adult men with chronic lymphocytic leukaemia arising within a week of completion of treatment with fludarabine. In all cases, withdrawal of fludarabine and treatment of the blistering was associated with marked cutaneous improvement. Fludarabine, a synthetic nucleoside analogue, which has only been available in Britain since 1994, is known to be associated with autoimmune phenomena and may have been involved in the development of paraneoplastic pemphigus in these cases. PMID- 11422054 TI - Topical 5-aminolaevulinic acid photodynamic therapy for tumour-stage mycosis fungoides. PMID- 11422055 TI - Topical application of mycophenolate mofetil in plaque-type psoriasis. PMID- 11422056 TI - Vitiligo following narrow-band TL-01 phototherapy for psoriasis. PMID- 11422057 TI - Repigmentation of localized vitiligo with the xenon chloride laser. PMID- 11422058 TI - Pulsed dye laser treatment for inflammatory linear verrucous epidermal naevus. PMID- 11422059 TI - Port-wine stains and eczema. PMID- 11422060 TI - Intact nerve supply for eczema. PMID- 11422062 TI - Cutaneous inflammatory pseudotumour (plasma cell granuloma). PMID- 11422063 TI - Subcutaneous cellular neurothekeoma: a pseudosarcomatous tumour. PMID- 11422064 TI - A case of fibro-osseous pseudotumour of the digits. PMID- 11422065 TI - Post-inflammatory depigmentation following allergic contact dermatitis to chloroxylenol. PMID- 11422066 TI - Propolis-induced granulomatous contact dermatitis accompanied by marked lymphadenopathy. PMID- 11422067 TI - Endothelial cells express an alpha-melanocyte-stimulating hormone-like immunoreactivity in prurigo nodularis. PMID- 11422068 TI - Progressive macular leucoderma in a patient with congenital ichthyosiform erythroderma. PMID- 11422069 TI - Suppurative panniculitis associated with alpha 1-antitrypsin deficiency (PiSZ phenotype) treated with doxycycline. PMID- 11422070 TI - Coma associated with scleromyxoedema and interferon alfa therapy. Full recovery after steroids and cyclophosphamide combined with plasmapheresis. PMID- 11422071 TI - Kindler syndrome complicated by squamous cell carcinoma of the hard palate: successful treatment with high-dose radiation therapy and granulocyte-macrophage colony-stimulating factor. PMID- 11422072 TI - Burning fingers, but where is the fire? PMID- 11422073 TI - Early use of percutaneous endoscopic gastrostomy feeding in toxic epidermal necrolysis. PMID- 11422074 TI - Non-pigmenting fixed drug eruption: a new case due to eperisone hydrochloride. PMID- 11422075 TI - Extensive fixed drug eruption due to lamotrigine. PMID- 11422076 TI - Fixed drug eruption due to naproxen; lack of cross-reactivity with other propionic acid derivatives. PMID- 11422077 TI - Thalidomide-induced amenorrhoea: two cases. PMID- 11422078 TI - Discoloured tongue: a new cause? PMID- 11422079 TI - Enzymes on microbial pathogens and Trichomonas vaginalis: molecular mimicry and functional diversity. PMID- 11422080 TI - Fluorescent Plasmodium berghei sporozoites and pre-erythrocytic stages: a new tool to study mosquito and mammalian host interactions with malaria parasites. AB - To track malaria parasites for biological studies within the mosquito and mammalian hosts, we constructed a stably transformed clonal line of Plasmodium berghei, PbFluspo, in which sporogonic and pre-erythrocytic liver-stage parasites are autonomously fluorescent. A cassette containing the structural gene for the FACS-adapted green fluorescent protein mutant 2 (GFPmut2), expressed from the 5' and 3' flanking sequences of the circumsporozoite (CS) protein gene, was integrated and expressed at the endogenous CS locus. Recombinant parasites, which bear a wild-type copy of CS, generated highly fluorescent oocysts and sporozoites that invaded mosquito salivary glands and were transmitted normally to rodent hosts. The parasites infected cultured hepatocytes in vitro, where they developed into fluorescent pre-erythrocytic forms. Mammalian cells infected by these parasites can be separated from non-infected cells by fluorescence activated cell sorter (FACS) analysis. These fluorescent insect and mammalian stages of P. berghei should be useful for phenotypic studies in their respective hosts, as well as for identification of new genes expressed in these parasite stages. PMID- 11422081 TI - Burkholderia pseudomallei kills the nematode Caenorhabditis elegans using an endotoxin-mediated paralysis. AB - We investigated a non-mammalian host model system for fitness in genetic screening for virulence-attenuating mutations in the potential biowarfare agents Burkholderia pseudomallei and Burkholderia mallei. We determined that B. pseudomallei is able to cause 'disease-like' symptoms and kill the nematode Caenorhabditis elegans. Analysis of killing in the surrogate disease model with B. pseudomallei mutants indicated that killing did not require lipopolysaccharide (LPS) O-antigen, aminoglycoside/macrolide efflux pumping, type II pathway secreted exoenzymes or motility. Burkholderia thailandensis and some strains of Burkholderia cepacia also killed nematodes. Manipulation of the nematode host genotype suggests that the neuromuscular intoxication caused by both B. pseudomallei and B. thailandensis acts in part through a disruption of normal Ca2+ signal transduction. Both species produce a UV-sensitive, gamma-irradiation resistant, limited diffusion, paralytic agent as part of their nematode pathogenic mechanism. The results of this investigation suggest that killing by B. pseudomallei is an active process in C. elegans, and that the C. elegans model might be useful for the identification of vertebrate animal virulence factors in B. pseudomallei. PMID- 11422082 TI - Cytochrome c-mediated caspase-9 activation triggers apoptosis in Streptococcus pyogenes-infected epithelial cells. AB - Epithelial cells are the initial sites of host invasion by group A Streptococcus pyogenes (GAS), and their infection of epithelial cells has been suggested to induce apoptosis. However, the mechanism responsible for bacteria-host interaction and the induction of apoptosis has not been clearly understood. We demonstrate here that human pharyngeal epithelial HEp-2 cells became apoptotic with DNA fragmentation by invasion of GAS strains JRS4 (M6+, F1+) and JRS145 (M6 , F1+ mutant of JRS4), whereas apoptotic cellular changes were not observed in SAM1 (M6+, F1- mutant) or SAM2 (M6-, F1- mutant) infected HEp-2 cells. Confocal microscopy revealed that Bax translocation to mitochondria and cytochrome c release occurred after 4 h of infection. Western blot analyses showed that the amounts of Bcl-2 and Bcl-xL were decreased in the mitochondria of infected cells. In addition, we demonstrated that the release of nuclear histone from infected cells was prevented by the addition of caspase-9 inhibitor (Ac-LEHD-CHO). We conclude that the internalization of GAS in epithelial cells is necessary and sufficient for the induction of apoptosis, which is initiated by mitochondrial dysfunction, and the mechanism of GAS-induced apoptosis is clearly different from that induced by other intracellular invasive bacteria, e.g. Shigella and Salmonella species. PMID- 11422083 TI - Unusual intracellular trafficking of Salmonella typhimurium in human melanoma cells. AB - Salmonella spp. are enterobacteria capable of invading and replicating in both professional and non-professional phagocytes. Here, we investigate the fate of S. typhimurium in human melanoma MelJuSo cells. The bacterium entered MelJuSo cells by a trigger mechanism and resided within a unique organelle, the Salmonella containing vacuole (SCV). The SCV acquired early endosomal markers transiently and then underwent a series of membrane modifications. In HeLa cells, vacuole maturation is characterized by the simultaneous acquisition of the lysosomal membrane glycoproteins (Lgps) Lamp1, CD63 and vacuolar (v)-ATPase; in MelJuSo cells, however, acquisition of CD63 and v-ATPase preceded that of Lamp1. A very striking event in MelJuSo cells was the arrest of bacterial septation starting from 8 h after infection. Bacteria nevertheless continued to elongate, remained morphologically intact and viable and were eventually exocytosed. This original feature was observed in several skin-related cells including melanocytes, suggesting that it may provide the basis for an efficient host defence mechanism against Salmonella infection. PMID- 11422084 TI - Role of sipA in the early stages of Salmonella typhimurium entry into epithelial cells. AB - Salmonella virulence depends on an ability to invade host cells, which is in turn dependent on a type III protein secretion system encoded in Salmonella pathogenicity island 1 (SPI1). Several protein targets of the SPI1-encoded secretion system are translocated into host cells, where they subvert cellular processes that contribute to bacterial invasion, actin rearrangement, membrane ruffling and other aspects of virulence. We examined the role of sipA (encoding the translocated protein SipA) and found that a sipA mutant was significantly less invasive in Madin-Darby canine kidney (MDCK) cells than in its parental strain at the earliest stages of infection (5 min). The invasion defect associated with sipA was no longer apparent after 15 min of infection. Confocal microscopy of F-actin in tetramethyl rhodamine isothiocyanate (TRITC)-phalloidin stained MDCK cells revealed no difference in either the frequency or the morphology of membrane ruffles induced by wild-type and sipA mutant strains of S. typhimurium. Time-lapse phase-contrast microscopy of membrane ruffle propagation in live cells confirmed that the sipA mutant induced membrane ruffles as efficiently as the wild-type bacteria. These studies also revealed that, after ruffle propagation, individual sipA mutant S. typhimurium either invaded more slowly than wild-type bacteria or failed to invade at all. Furthermore, although wild-type S. typhimurium typically maintained a position central to the developing membrane ruffle, sipA mutant bacteria frequently moved initially to the periphery of the spreading ruffle and were sometimes observed to detach from it. A wild-type pattern of invasion was restored to the sipA mutant after the introduction of sipA on a plasmid. Together, these data indicate that loss of sipA significantly decreases the efficiency of S. typhimurium invasion at the early stages of infection without affecting its ability to induce membrane ruffles. It thus appears that the secreted effector protein SipA promotes invasion by a previously unrecognized mechanism separate from the induction of membrane ruffling per se. PMID- 11422085 TI - Low iron availability modulates the course of Chlamydia pneumoniae infection. AB - Chlamydiae are obligate intracellular bacteria residing exclusively in host cell vesicles termed inclusions. We have investigated the effects of deferoxamine mesylate (DAM)-induced iron deficiency on the growth of Chlamydia pneumoniae and Chlamydia trachomatis serovar L2. In epithelial cells subjected to iron starvation and infected with either C. pneumoniae or C. trachomatis L2, small inclusions were formed, and the infectivity of chlamydial progeny was impaired. Moreover, for C. trachomatis L2, we observed a delay in homotypic fusion of inclusions. The inhibitory effects of DAM were reversed by adding exogenous iron saturated transferrin, which restored the production of infectious chlamydiae. Electron microscopy examination of iron-deprived specimens revealed that the small inclusions contained reduced numbers of C. pneumoniae that were mostly reticulate bodies. We have previously reported specific accumulation of transferrin receptors (TfRs) around C. pneumoniae inclusions within cells grown under normal conditions. Using confocal and electron microscopy, we show here a remarkable increase in the amount of TfRs surrounding the inclusions in iron starved cultures. It has been shown that iron is an essential factor in the growth and survival of C. trachomatis. Here, we postulate that, for C. pneumoniae also, iron is an indispensable element and that Chlamydia may use iron transport pathways of the host by attracting TfR to the phagosome. PMID- 11422086 TI - Ophthalmoplegic migraine. PMID- 11422087 TI - SUNCT syndrome or trigeminal neuralgia with lacrimation. PMID- 11422088 TI - Ophthalmoplegic migraine: a recurrent demyelinating neuropathy? AB - The demonstration by magnetic resonance imaging (MRI) scanning of thickening and enhancement of the cisternal part of the oculomotor nerve in patients diagnosed as "ophthalmoplegic migraine" prompts reconsideration of this uncommon disorder. The case histories of five patients, three male and two female, varying in age from 6 to 30 years, are presented here. Recurrent painful ophthalmoplegia started in infancy in two cases, childhood in two instances and adult life in one. One child had his first attacks at 3, 5 and 12 months of age, on each occasion 10 days after an injection of triple vaccine. The possibility of this condition being a recurrent demyelinating neuropathy is considered and its possible relationship to migraine explored. PMID- 11422089 TI - Intravenous metamizol (Dipyrone) in acute migraine treatment and in episodic tension-type headache--a placebo-controlled study. AB - Acute headache is a very frequent symptom, responsible for significant demand at primary care units and emergency rooms. In such sets in Brazil, metamizol is easily found but, on the other hand, neither ergotics nor triptans are available. The aim of this study is to compare intravenous metamizol with placebo in the acute treatment of migraine with aura, migraine without aura and episodic tension type headache. Fifty-four migraine with aura patients, 95 migraine without aura patients and 30 tension-type headache patients were treated with metamizol. Ninety patients (30 migraine with aura, 30 migraine without aura and 30 tension type headache patients) received placebo. Pain intensity, nausea, aura, photo- and phonophobia were investigated at 30 min and 60 min after the administration of the drug. Significant improvement of pain after 30 min and 60 min post-dosage was achieved from metamizol groups compared with placebo groups. Significant improvement of all other symptoms was achieved after 60 min post-dosage. Side effects were mild and with small incidence. Metamizol is an effective, safe and low price drug. It may be regarded as a good alternative drug for the treatment of common acute primary headaches. PMID- 11422090 TI - Neuropeptide localization in the "migraine generator" region of the human brainstem. AB - Evidence from animals and humans suggests that brainstem nuclei such as the raphe nuclei, the locus coeruleus (LC) and the periaqueductal grey matter (PAG), are involved in the pathophysiology of migraine. In order to understand possible neurotransmitters involved we have, by means of indirect immunocytochemistry, analysed these regions for the occurrence and distribution of calcitonin gene related peptide (CGRP), substance P (SP), pituitary adenylate-cyclase activating peptide (PACAP) and vasoactive intestinal polypeptide (VIP). CGRP-immunoreactive (-ir) cell bodies, but no fibres, were found to occur in high numbers, constituting 80% of all nerve cell bodies in the LC. A smaller number of these nerve cell bodies (40%) in the LC proved to be PACAP-ir. The LC neurones also stored the vesicular monoamine transporter (VMAT)- and the C-terminal flanking peptide of neuropeptide Y (C-PON)-ir, illustrating their adrenergic nature. Double immunostaining revealed that all VMAT-and C-PON-containing neurones, in addition, stored CGRP. Immunoreactive cell bodies were not seen in the nucleus raphe magnus (NRM) or PAG. Numerous SP-ir nerve fibres were observed in the NRM, the LC and the PAG. Few PACAP-ir nerve fibres were detected in the PAG and few VIP-ir nerve fibres were seen in the NRM and the PAG. PMID- 11422091 TI - Lanepitant, an NK-1 antagonist, in migraine prevention. AB - Lanepitant, a potent non-peptide neurokinin-1 receptor antagonist, inhibits neurogenic dural inflammation, and may have a role in migraine therapy. This study evaluated the effect of lanepitant taken daily for migraine prevention. Patients with migraine headaches with and without aura by International Headache Society classification criteria were enrolled in a 12-week double-blind, parallel design study comparing the effect of 200 mg qd lanepitant (n = 42) and placebo (n = 42) on reduction of migraine frequency. The primary outcome measure was response rate, i.e. the proportion of patients with a 50% reduction in days of headache. Of the 84 patients enrolled, 90.5% were female. The endpoint response rate for lanepitant-treated patients (41.0%) was not statistically significantly (P = 0.065) greater than that for placebo-treated patients (22.0%). No efficacy variables differed significantly between treatments, except for response rates at month 3 (P = 0.045). Higher plasma concentrations were no more effective than lower concentrations. In this study lanepitant was not effective in preventing migraine, but was well tolerated. These results do not support a role for NK-1 antagonism in migraine prevention. PMID- 11422092 TI - Referred pain after painful stimulation of the greater occipital nerve in humans: evidence of convergence of cervical afferences on trigeminal nuclei. AB - Cranial sensory innervation is supplied mainly by the trigeminal nerves and by the first cervical nerves. Excitatory and inhibitory interactions among those nerve roots may occur in a mechanism called nociceptive convergence, leading to loss of somato-sensory spatial specificity. Three volunteers in an experimental trial had sterile water injected over their greater occipital nerve on one side of the neck. Pain intensity was evaluated 10, 30 and 120 s after the injection. Two of the patients reported intense pain. Trigeminal autonomic features, suggestive of parasympathetic activation, were seen associated with trigeminally distributed pain. These data add to and reinforce previous evidence of convergence of cervical afferents on the trigeminal sensory circuit. PMID- 11422093 TI - Alpha1-adrenoceptor subtypes mediating vasoconstriction in the carotid circulation of anaesthetized pigs: possible avenues for antimigraine drug development. AB - It has recently been shown that the alpha-adrenoceptors mediating vasoconstriction of porcine carotid arteriovenous anastomoses resemble both alpha1- and alpha2-adrenoceptors, but no attempt was made to identify the specific subtypes (alpha1A, alpha1B and alpha1D) involved. Therefore, the present study was designed to elucidate the specific subtype(s) of alpha1-adrenoceptors involved in the above response, using the alpha1-adrenoceptor agonist phenylephrine and alpha1-adrenoceptor antagonists 5-methylurapidil (alpha1A), L 765 314 (alpha1B) and BMY 7378 (alpha1D). Ten-minute intracarotid infusions of phenylephrine (1, 3 and 10 microgkg-1.min-1) induced a dose-dependent decrease in total carotid and arteriovenous anastomotic conductance, accompanied by a small tachycardia. These carotid vascular effects were abolished by L-765 314 (1000 microgkg-1; i.v.), while these responses were only attenuated by 5-methylurapidil (1000 microgkg-1; i.v.), and BMY 7378 (1000 microgkg-1; i.v.). Furthermore, intravenous bolus injections of phenylephrine (3 and 10 microgkg-1) produced a dose-dependent vasopressor response, which was only affected by 1000 microgkg-1 of 5-methylurapidil, while the other antagonists were ineffective. These results, coupled to the binding affinities of the above antagonists at the different alpha1-adrenoceptors, suggest that both alpha1A- and alpha1B-adrenoceptors mediate constriction of carotid arteriovenous anastomoses in anaesthetized pigs. In view of the less ubiquitous nature of alpha1B- compared to alpha1A adrenoceptors, the development of potent and selective alpha1B-adrenoceptor agonists may prove to be important for the treatment of migraine. PMID- 11422094 TI - A comparative study of oral acetylsalicyclic acid and metoprolol for the prophylactic treatment of migraine. A randomized, controlled, double-blind, parallel group phase III study. AB - This study was a multinational, multicentre, double-blind, active controlled phase III trial designed to investigate efficacy and safety of 300 mg acetylsalicyclic acid (ASA) (n = 135) vs. 200 mg metoprolol (n = 135) in the prophylaxis of migraine. In total 270 (51 male and 219 female) patients, aged 18 65 years, suffering between two and six migraine attacks per month were recruited. The main objective was to show equivalence with respect to efficacy, defined as a 50% reduction in the rate of migraine attacks. A run-in phase was carried out with placebo for 4 weeks, followed by a 16-week drug phase. In both treatment groups the median frequency of migraine attacks improved during the study period, from three to two in the ASA group and from three to one in the metoprolol group; 45.2% of all metoprolol patients were responders compared with 29.6% with ASA. Medication-related adverse events were less frequent in the ASA group (37) than in the metoprolol group (73). The findings from this trial show that metoprolol is superior to ASA for migraine prophylaxis but has more side effects. Acetylsalicylic acid is better tolerated than metoprolol. Using a strict responder criterion ASA showed a responder rate comparable with the placebo rate in the literature. PMID- 11422095 TI - Meta-analysis of rizatriptan efficacy in randomized controlled clinical trials. AB - Data from seven randomized, placebo-controlled, double-blind phase III clinical trials were analysed to further evaluate the efficacy of rizatriptan 10 mg (n = 2068) in comparison with placebo (n = 1260) and rizatriptan 5 mg (n = 1486) for the acute treatment of a migraine attack. Migraine was diagnosed according to International Headache Society criteria. Headache severity, associated migraine symptoms and functional disability were measured immediately before dosing and at 0.5, 1, 1.5 and 2 h. Headache recurrence (return of moderate or severe headache after an initial response) was also recorded. In addition to conventional pain relief (reduction of moderate or severe headache to mild or none) and pain free measures, the analysis looked at the elimination of associated migraine symptoms and disability in patients who had symptoms or disability at baseline. Maintenance of pain relief or pain-free status over 24 h was also analysed. At 2 h, rizatriptan 10 mg was significantly more effective than placebo for pain relief (71% vs. 38%, P < 0.001), and for elimination of pain, nausea, photophobia, phonophobia and functional disability. The benefit was maintained over 24 h; 37% of patients on rizatriptan 10 mg had sustained pain relief vs. 18% for placebo (P < 0.001). Rizatriptan 10 mg was also more effective than rizatriptan 5 mg, with a significant superiority at 2 h on all measures except for elimination of nausea. The benefit was maintained over 24 h; 38% of patients on rizatriptan 10 mg had sustained pain relief vs. 32% for rizatriptan 5 mg (P = 0.001). PMID- 11422096 TI - Evidence for activation of the coagulation system in migraine with aura. AB - Migraine with aura has been shown to be an independent risk factor for stroke. Although the precise mechanism of migraine-related stroke is not known, risk factors for hypercoagulability have been found in migraineurs. Prothrombin factor 1.2 (F1.2) is a cleavage product of prothrombin. Elevated plasma F1.2 has been shown to be a sensitive and a specific marker of ongoing thrombin generation, and thus may serve as an indicator of hypercoagulability. In this study we determined plasma F1.2 levels in 35 patients with migraine (22 with aura and 13 without aura) and in 24 healthy age- and sex-matched volunteers. Elevated F1.2 levels were found in 11 of 22 (50%) patients with migraine with aura (1.25-3.5 nmol/l). None of the patients with migraine without aura nor any of the healthy volunteers had elevated plasma F1.2 levels (normal < 1.1 nmol/l). We conclude that prothrombin F1.2 levels are elevated in a significant number of patients with migraine with aura but not in patients with migraine without aura. This finding suggests that there is activation of the clotting system in certain patients with migraine with aura. PMID- 11422097 TI - Treatment of migraine attacks with intranasal alniditan: an open study. AB - In this open phase-II clinical tolerability trial 17 neurologists enrolled a total of 112 patients and instructed them to administer a maximum of two doses of intranasal alniditan, a 5-HT1B/D receptor agonist, for the treatment of three consecutive migraine attacks of moderate to severe intensity. A second dose of the trial medication was allowed within 1-24 h after the first administration. At 1 h after intranasal administration, 70/103 (68%) patients had responded to treatment (reduction from severe or moderate headache before treatment to mild or no headache) after their first migraine attack, 65/94 (69%) after their second and 52/75 (71%) after their third. In 187/270 (69%) of all attacks, patients were considered responders at 1 h. The median time to onset of effect was 30 min. The migraine headache recurred in 44% (attack 1), 55% (attack 2) and 44% (attack 3) after 4-5 h. Sixty-eight per cent of the patients reported nasal irritation, 19% taste disturbance and 44% throat irritation. Alniditan 2 mg, administered via the intranasal route, was effective in relieving migraine headaches in over two thirds of the patients at 1 h. PMID- 11422098 TI - Application of the International Headache Society classification criteria in 652 cluster headache patients. AB - We applied the International Headache Society (IHS) classification coding parameters to a study population of 652 cluster headache (CH) patients, in order to determine how many patients did not fulfil the diagnostic criteria for group 3.1 and to find out any diagnostic elements that could be changed in the upcoming revision of the classification to make it more relevant to current clinical practice. Ninety-nine patients were found to have cluster-like disorder (3.3), including 74 (74.7%) who did not fulfil the diagnostic criteria for CH, because either pain was not associated with any of the accompanying autonomic phenomena listed in the classification or it was not located orbitally, supraorbitally and/or temporally. A review of our total sample showed that 72.0% of patients reported frontal and occipital pain location; in 61.8%, 33.4% and 39.1% of cases, attacks were also accompanied by restlessness/agitation, nausea and photophobia, respectively. In a coding system that took into account the diagnostic elements that we considered in our study, group 3.1 of the existing IHS classification would actually include 51 of the 99 patients currently coded as 3.3. PMID- 11422099 TI - SUNCT syndrome or trigeminal neuralgia with lacrimation and conjunctival injection? PMID- 11422100 TI - SUNCT syndrome: a treatment option with local opioid blockade of the superior cervical ganglion? A case report. PMID- 11422101 TI - SUNCT syndrome and seborrheic dermatitis associated with craneosynostosis. PMID- 11422102 TI - Thyroid cancer in the UK: can we do it better? PMID- 11422103 TI - Diversity in developmental actions of androgens. PMID- 11422104 TI - Body composition and quality of life in adults with growth hormone deficiency; effects of low-dose growth hormone replacement. AB - OBJECTIVE: Adult growth hormone deficiency (AGHD) is characterized by abnormalities in body composition and a poor perceived quality of life (QoL). Weight-based high-dose growth hormone replacement (GHR) results in improvements in body composition and QoL in AGHD. However, a high patient percentage reported side-effects on high-dose GHR resulting in a high rate of patient withdrawal from growth hormone (GH) treatment. High-dose GH therapy also leads to supraphysiological serum insulin-like growth factor-I (IGF-I) concentrations that have been associated with breast and prostate cancer, raising major concerns over the use of such high-dose GH regimen in AGHD. The aim of this study was to assess the effects of low-dose growth hormone replacement (GHR) on body composition and QoL as early as 1 and 3 months. STUDY DESIGN: A prospective, open treatment design study to determine the early effects of low-dose GH administration on body composition and QoL. GH was initiated at a daily dose of 0.4-0.5 IU, and titrated up to achieve and maintain IGF-I standard deviation score (IGF-I SDS) between the median and upper end of the age-related reference range. PATIENTS: Forty-six, post-pituitary surgery, severe AGHD patients (22 women), defined as peak GH response < 9 mU/l to provocative testing. The mean age was 50.4 years (range 26 72). Forty-three patients required additional pituitary replacement hormones following pituitary surgery and were on optimal doses at recruitment. MEASUREMENTS: Body composition and QoL were assessed prior to GHR and subsequently at 1 and 3 months after initiating GHR. Body mass index (BMI) and waist hip ratio (WHR) were calculated from measurements of height, weight, and waist and hip circumference, respectively. Bioelectrical impedance analysis (BIA) was used to determine body fat and lean body mass. QoL was assessed using the disease-specific 'QoL-assessment of growth hormone deficiency in adults (QoL AGHDA)' questionnaire. Serum IGF-I was measured at each visit to assess the adequacy of GHR. RESULTS: IGF-I and IGF-I SDS increased significantly at 1 and 3 months (P < 0.001) after commencing GHR. The increase in IGF-I (P < 0.05) and IGF I SDS (P < 0.01) was significant between 1 and 3 months in the absence of any significant increase in GH dose (P = ns) during this period. Eighty-five per cent of patients achieved IGF-I SDS levels between median and upper end of the age related reference range after 3 months of GHR, and no side-effects were reported during this period. There was a significant reduction in body fat percentage (BFP) from 36.1 +/- 9.1% at baseline to 34.9 +/- 9.3% (P < 0.01) at 1 month and 34.1 +/- 9.2% (P < 0.001) at 3 months. Body fat mass (BFM) reduced from 32.8 +/- 13.6 kg at baseline to 31.9 +/- 13.9 kg at 1 month (P < 0.05) and 31.1 +/- 13.6 kg at 3 months (P < 0.001). These changes in BFP and BFM occurred in the absence of any significant change in BMI and WHR (P = ns). Lean body mass (LBM) was 55.9 +/- 11.1 kg at baseline and increased to 57.1 +/- 11.3 kg after 1 month (P < 0.01) and to 57.6 +/- 11.5 kg (P < 0.001) after 3 months of GHR. Significant improvement was observed in the perceived QoL with the AGHD assessment scores reducing from 13.3 +/- 6.4 to 11.5 +/- 6.6 within 1 month (P < 0.01) and 10.0 +/- 6.6 at 3 months (P < 0.001). There was no significant correlation between improvement in QoL and changes in body fat percentage (r = 0.01 at 1 month and r = 0.12 at 3 months, P = ns) or IGF-I levels (r = 0.04 and r = 0.003, P = ns at 1 and 3 months, respectively). The improvement in body composition and QoL was significant between 1 and 3 months. CONCLUSIONS: Low-dose GHR improves body composition and QoL as early as 1 month after commencement and the beneficial effects continue at 3 months. Most importantly, these changes occur in the absence of side-effects. We therefore suggest the use of low-dose GH therapy, maintaining IGF-I between the median and upper end of the age-related reference range, for the treatment of AGHD. PMID- 11422105 TI - An audit of management of differentiated thyroid cancer in specialist and non specialist clinic settings. AB - OBJECTIVE: Thyroid cancer is the most common endocrine malignancy but is none the less rare. Some aspects of its management remain controversial. Previous audits of patient management in the United Kingdom have revealed deficiencies, especially in communication between specialists. We have audited patient management in a large university-associated teaching hospital, assessing points of good practice identified from published guidelines and reviews, and have compared findings in groups of patients managed jointly by specialists with an interest in thyroid cancer (including surgeon, endocrinologist and oncologist) with a group managed by other clinicians outside that setting. DESIGN AND PATIENTS: Retrospective case-note review of 205 patients with differentiated (papillary or follicular) cancer including group A (n = 134; managed in a specialist multi-disciplinary clinic setting) and group B (n = 71; managed in other clinic settings). Points of good practice investigated were adequacy of surgery, surgical complications, prescription and adequacy of T4 treatment, adequacy of monitoring by measurement of serum thyroglobulin and action taken and appropriate administration of ablative radioiodine. RESULTS: Deficiencies in management of the cohort as a whole were identified, including inadequate surgery and inadequate TSH suppression in approximately one-fifth of the cases. Monitoring with thyroglobulin measurements and action when serum thyroglobulin was high were also inadequate in some cases and ablative radioiodine was not given, despite being indicated in 11.7% of the cohort. Inadequate surgery and failure to administer radioiodine were less common in those managed in a specialist clinic setting than in those managed in other clinic settings. CONCLUSIONS: The findings highlight the need for locally agreed protocols in managing relatively rare endocrine disorders such as thyroid cancer and argue in favour of centralization of expertise and patient management in multi disciplinary specialist clinic settings. PMID- 11422106 TI - Polymorphic differences from normal in the aldosterone synthase gene (CYP11B2) in patients with primary hyperaldosteronism and adrenal tumour (Conn's syndrome). AB - OBJECTIVE: The hypertension of Conn's syndrome is due to autonomous aldosterone production by a unilateral adrenocortical adenoma. The source of tumour initiation and the reasons for excess aldosterone production as opposed to cortisol are not known, although variations in the promoter region of the gene coding for aldosterone synthase (CYP11B2) might account for the altered rate of aldosterone secretion. DESIGN: In a series (n = 27) of well-characterized Conn's syndrome cases, the aldosterone synthase gene (CYP11B2) was screened by single strand conformational polymorphism (SSCP) for differences from the consensus sequence. RESULTS: No new mutations were found. The frequencies of two previously described linked polymorphisms, one a change of -344C to T in a putative steroidogenic factor-1 (SF-1) binding site and the other an exchange of intron 2 for that of CYP11B1 (conversion) were measured in tumour and genomic DNA. The frequency of the SF-1 T allele (P < 0.0001) and the conversion allele (P < 0.001) were markedly different between the Conn's syndrome group and the normal controls. However, the frequency did not differ between tumour and genomic DNA in the patient group. CONCLUSION: While it is unlikely that this difference from normal is related to tumour growth, these genotypes may predispose the tumour to aldosterone production. PMID- 11422107 TI - Decreased dopaminergic tone and increased basal bioactive prolactin in men with human immunodeficiency virus infection. AB - OBJECTIVE: The aims of the study were: (1) to assess dopaminergic tone in a group of HIV infected men and the bioactivity and the molecular species of their circulating PRL in comparison with healthy men and (2) to search for a correlation between serum PRL and CD4+ T lymphocytes and viral load. DESIGN: In a cross-sectional study the effect of acute dopaminergic blockade with intravenous metoclopramide on serum PRL (both immunoreactive and biologically active), TSH and PRL circulating molecular isoforms was evaluated. PATIENTS: Twenty untreated HIV infected men category C2 or C3, mean (SD) age 26.9 (6.3) years, were compared to 14 clinically healthy HIV-negative men, age 25.4 (2.3) years. MEASUREMENTS: Under fasting conditions and following metoclopramide administration duplicate measurements of serum immunoreactive PRL, bioactive PRL (PRL dependent Nb2 lymphoma cell assay) and immunoreactive TSH were performed. The molecular species of circulating PRL were determined by immunoblot analysis, CD4+ T lymphocytes by flow cytometry and the viral load using a nucleic acid sequence-based amplification assay. RESULTS: In HIV infected men fasting bioactive (but not immunoreactive) PRL was higher (P = 0.03), but the stimulated PRL (both immunoreactive and bioactive) was lower than in healthy men throughout the test (P < or = 0.01). Fasting serum TSH was similar in HIV-infected and healthy men while its response to metoclopramide was absent in the former but not in the latter (P = 0.049). A 23.5-kD PRL was the predominant circulating isoform both in patients and healthy men. Considering HIV-infected and healthy men, CD4+ T lymphocytes correlated negatively with fasting bioactive PRL (P = 0.008) and positively with the area under the PRL (both immunoreactive and bioactive) curves (P < 0.001). The viral load was negatively correlated with the area under the curve of the bioactive/immunoreactive ratio (P = 0.008). CONCLUSIONS: The raised fasting bioactive PRL, the diminished response of both immunoreactive and bioactive PRL and the absent TSH response to metoclopramide in HIV infected men, suggest the existence of a decreased, but not absent dopaminergic tone. A monomeric form of PRL was the predominant circulating species, as in healthy men, and this hormone seems to be associated both with CD4+ T lymphocytes and the viral load. PMID- 11422108 TI - Effects of transdermal testosterone gel on bone turnover markers and bone mineral density in hypogonadal men. AB - OBJECTIVE: Androgen replacement has been reported to increase bone mineral density (BMD) in hypogonadal men. We studied the effects of 6 months of treatment with a new transdermal testosterone (T) gel preparation on bone turnover markers and BMD. DESIGN: This was a prospective, randomized, multicentre, parallel clinical trial where 227 hypogonadal men, mean age 51 years (range: 19-68 years) were studied in 16 academic and research institutions in the USA. Subjects were randomized to apply 1% T gel containing 50 or 100 mg T (delivering approximately 5-10 mg T/day) or two T patches (delivering 5 mg T/day) transdermally for 90 days. At day 91, depending on the serum T concentration, the T gel dose was adjusted upward or downward to 75 mg T/day until day 180. No dose adjustment occurred in the T patch group. MEASUREMENTS: Serum T, free T and oestradiol, bone turnover markers and BMD were measured on days 0, 30, 90 and 180 before and after treatment. RESULTS: Application of T gel 100 mg/day resulted in serum T concentrations 1.4 and 1.9-fold higher than in the T gel 50 mg/day and the T patch groups, respectively. Proportional increases occurred in serum oestradiol. Urine N-telopeptide/creatinine ratio, a marker for bone resorption, decreased significantly (P = 0.0019) only in the T gel 100 mg/day group. Serum bone osteoblastic activity markers (osteocalcin, procollagen and skeletal alkaline phosphatase) increased significantly during the first 90 days of treatment without intergroup differences but declined to baseline thereafter. BMD increased significantly both in the hip (+1.1 +/- 0.3%) and spine (+2.2 +/- 0.5%) only in the T gel 100 mg/day group (P = 0.0001). CONCLUSIONS: Transdermal testosterone gel application for 6 months decreased bone resorption markers and increased osteoblastic activity markers for a short period, which resulted in a small but significant increase in BMD. Ongoing long-term studies should answer whether the observed increases in BMD are sustained or continue to be dependent on the dose of testosterone administered. PMID- 11422109 TI - Genetic analysis of the cytochrome P-450c17alpha (CYP17) and aldosterone synthase (CYP11B2) in Japanese patients with 17alpha-hydroxylase deficiency. AB - OBJECTIVE: To determine the clinical and molecular genetic characterization of two Japanese patients with 17alpha-hydroxylase deficiency, we analysed the 17alpha-hydroxylase/17,20-lyase gene (CYP17). Next, to clarify the mechanism of hypoaldosteronism in 17alpha-hydroxylase deficiency, we analysed the expression of aldosterone synthase (CYP11B2) messenger RNA and sequenced CYP11B2 in these patients. PATIENTS: Patient 1 (46 XY), phenotypically female, sought medical attention for hypertension, amenorrhea and infantile genitalia. Patient 2 (46 XX), phenotypically female, presented for hypertension and amenorrhea. Hormonal data in both patients showed decreased levels of sex steroids, cortisol, aldosterone and plasma renin activity and extreme elevation of deoxycortisol. DESIGN: Direct sequencing of CYP17 and CYP11B2 was performed using genomic DNA from the patients. An expression studies of mutated forms of CYP17 was performed using COS-1 cells. The expression of CYP11B2 messenger RNA in mononuclear leucocytes (MNLs) of these patients and normal subjects was measured using the competitive polymerase chain reaction METHOD: The effect of renin secretion stimulation on the levels of CYP11B2 messenger RNA in MNLs of normal subjects was also studied. RESULTS: We detected two novel genetic defects in 17alpha hydroxylase. Sequence analysis revealed one base pair deletion (T) at codon 243 in exon 4 in patient 1. CYP17 in patient 2 contained a point mutation (C to T) at position 415 in exon 8. Transfected cells of mutant from patient 1 had no 17alpha hydroxylase or 17,20-lyase activity. The R415C mutant protein showed very weak activity of 17alpha-hydroxylase or 17,20-lyase activity. In the renin secretion stimulating test, the increase in CYP11B2 messenger RNA levels in MNLs was parallel with that of plasma aldosterone concentration. The expression of CYP11B2 mRNA in NMLs of these patients was lower compared to controls. No mutations in CYP11B2, including the 5' flanking region, were found. CONCLUSIONS: These results indicate that the novel mutations of the CYP17 gene found in these patients inactivate cytochrome P450c17 function, and that hypoaldosteronism in these patients may be partly explained by a decreased activity of aldosterone synthase, which is regulated at the transcriptional level. PMID- 11422110 TI - Ghrelin and growth hormone (GH) secretagogue receptor (GHSR) mRNA expression in human pituitary adenomas. AB - OBJECTIVE: The level of growth hormone (GH), growth hormone secretogogue (GHS) and GHS receptor (GHSR) messenger ribonucleic acid (mRNA) expression has been reported as being higher in GH-producing pituitary adenomas than in other types of pituitary adenomas. Recently, ghrelin, an endogenous ligand specific for GHSR, was isolated. Therefore, we attempted to clarify whether ghrelin mRNA is expressed in various types of human pituitary adenoma by competitive reverse transcription-polymerase chain reaction (RT-PCR). We also examined the relationship between the levels of ghrelin or GHSR mRNA and hormonal and tumour characteristics in patients with pituitary adenomas. PATIENTS: Pituitary adenoma tissue was obtained at surgery from 13 patients with acromegaly, 4 with prolactinomas, 5 with gonadotrophin (Gn)-producing adenomas, 4 with non functioning adenomas, 2 with ACTH-producing adenomas and 2 with TSH-producing adenomas. METHODS: The expression levels of human ghrelin mRNA and GHSR mRNA were quantified using a competitive RT-PCR method. RESULTS: Ghrelin mRNA was detected in all pituitary adenoma tissues examined, with the highest mean level detected in non-functioning adenomas, a moderate level in GH-producing adenomas and Gn producing adenomas, and the lowest level in prolactinomas. The level of ghrelin mRNA expression in GH-producing adenomas correlated negatively with the size of the adenoma (n = 13) (r = - 0.756, P = 0.0028). Furthermore, the mean level of ghrelin mRNA expression in high-grade (III and IV of Hardy classification) GH producing adenomas was significantly lower than that in low-grade (I and II) GH producing adenomas (P = 0.0016). GHSR mRNA was also detected in all pituitary adenomas with the highest mean level in GH-producing adenomas, a moderate level in nonfunctioning adenoma, and the lowest level in prolactinoma and Gn-producing adenomas. CONCLUSIONS: Ghrelin mRNA, in addition to GHSR mRNA, is expressed in various types of pituitary adenoma with different levels of expression in each type. Our findings suggest that ghrelin produced in pituitary adenoma may play some role in the mechanism underlying the development of adenoma cells through autocrine and/or paracrine pathways. PMID- 11422111 TI - Hormone replacement therapy and risk of falling in early postmenopausal women - a population-based study. AB - OBJECTIVE: The purpose of this study was to examine the effect of hormone (oestrogen) replacement therapy (HRT) on the risk of falling among early postmenopausal women. METHODS: We assessed the incidence of falls in HRT users compared to non-users using population-based data from the Kuopio Osteoporosis Risk Factor and Prevention (OSTPRE) Study. The study group consisted of 9792 postmenopausal women who responded to the OSTPRE baseline and follow-up inquiries. RESULTS: A total of 3049 women reported sustaining a fall during the previous 12 months. The association between current continuous use of HRT and overall risk of falling was non-significant - 9% (P = 0.10). However, current continuous HRT use was associated with a decreased risk (- 30%) of non-slip falls (N = 1129) (P = 0.0001) but not with the risk (+ 9%) of slip falls (N = 1757) (P = 0.23). In early postmenopausal women (time since menopause < 5 years) the protective effect of current continuous HRT was strengthened: the risk of non slip falls was 71% lower in HRT users than non-users (P = 0.0035) if menopause had occurred within the past 2.5 years, and 43% lower (P = 0.0015) if time since menopause was 2.5-5 years. CONCLUSION: Hormone replacement therapy may reduce the risk of non-slip falls in early postmenopausal women. PMID- 11422112 TI - Increased serum inhibin B levels after varicocele treatment. AB - OBJECTIVE: Inhibin B is secreted by Sertoli cells in response to FSH and is the major feedback regulator of FSH secretion in man. The serum inhibin B level has emerged as a good marker of spermatogenesis and Sertoli cell function. Varicocele has been associated with infertility and disturbed spermatogenesis. We have studied the effect of varicocele treatment on serum inhibin B levels, with the aim of investigating the effect on spermatogenesis and the involvement of the Sertoli cell in varicocele pathophysiology. DESIGN AND PATIENTS: In a pre-post test design, the effect of varicocele surgery on inhibin B levels was studied in 30 infertile men. MEASUREMENTS: Endocrinology (inhibin B, FSH, LH, SHBG and testosterone) and semen analysis (sperm concentration, motility and morphology). RESULTS: In men receiving varicocele treatment, a significant increase in serum inhibin B levels was observed from 133.9 +/- 13.4 pretreatment to 167.8 +/- 16.1 ng/l after treatment (mean +/- SEM, P < 0.0001). No significant changes were observed in serum levels of FSH, LH and testosterone. The serum SHBG level decreased from 32.9 +/- 3.5 to 28.6 +/- 3.4 nmol/l (mean +/- SEM, P = 0.04) and the free androgen index was significantly increased from 66 +/- 5.9 pretreatment to 85 +/- 6.8 after treatment (P = 0.02, mean +/- SEM). Semen analysis showed a significant improvement in sperm concentration, from 6.5 +/- 1.9 pretreatment to 19.3 +/- 4.9 x 106/ml after treatment (P = 0.003, mean +/- SEM), and in sperm motility from the baseline level of 17 +/- 3 to 32 +/- 4% after treatment (P = 0.001, mean +/- SEM). CONCLUSIONS: Varicocele treatment can increase serum inhibin B levels, indicating improvement of spermatogenesis and Sertoli cell function. This finding suggests that the pathophysiology of varicocele involves impairment of Sertoli cell function or a different distribution of germ cell stages. PMID- 11422113 TI - The release of leptin and its effect on hormone release from human pituitary adenomas. AB - BACKGROUND: Leptin is the protein product of the obese gene, known to play an important role in body energy balance. The leptin receptor exists in numerous isoforms, the long isoform being the major form involved in signal transduction. Leptin expression has recently been demonstrated in the human pituitary, both in normal tissue and in pituitary adenomas. The long isoform of the leptin receptor has also been shown to be present in pituitary adenomas; however, contrasting results have been obtained regarding its expression in the normal human pituitary. AIM: The aim of this study was (i) to investigate the presence and pattern of distribution of leptin mRNA and the long isoform of its receptor mRNA in the normal pituitary and in different types of pituitary adenomas with RT-PCR; (ii) to study leptin secretion from human pituitary tumours in culture and (iii) to assess in vitro pituitary hormone release following stimulation with human leptin. RESULTS: Leptin receptor long isoform expression was detected in 2/4 GH secreting adenomas, 12/17 non-functioning adenomas, 5/9 ACTH-secreting adenomas, 1/2 prolactinomas, 2/2 FSH-secreting adenomas and 5/5 normal pituitaries. The receptor long isoform did not segregate with any particular tumour type, and varying levels of expression were detected between the tissues studied. Leptin mRNA was detected at a low level of expression in 2/7 GH-secreting adenomas, 9/14 non-functioning adenomas, 2/3 ACTH-secreting adenomas, 1/3 prolactinomas and 1/3 FSH-secreting adenomas. We were unable to detect leptin mRNA in any of the five normal pituitaries removed at autopsy; however, immunostaining of a non-tumorous pituitary adjacent to an adenoma removed at transsphenoidal surgery showed scattered leptin positive cells. Culture of pituitary adenomas showed that 16/47 released leptin into the incubation media. Leptin release did not correlate with tumour type or with any of the other pituitary hormones released. In vitro leptin stimulation of pituitary tumours caused stimulation of FSH and alpha-subunit secretion from a non-functioning adenoma and TSH secretion from a somatotroph adenoma. CONCLUSION: We conclude that not only is leptin stored within the pituitary, but it may also be released from pituitary cells and modulate other pituitary hormone secretion. Pituitary leptin may therefore be a novel paracrine regulator of pituitary function. PMID- 11422114 TI - Characterization of premature atherosclerosis of carotid arteries in acromegalic patients. AB - OBJECTIVE: Acromegalic patients have increased mortality from vascular diseases. Although atherosclerotic risk factors such as hypertension, diabetes mellitus and dyslipoproteinaemia are highly associated with acromegaly, the prevalence of premature atherosclerosis in acromegalic patients and its relationship to these risk factors have not been reported. DESIGN: We measured mean intima-media thickness (IMT) of the carotid arteries in 21 acromegalic patients without symptomatic atherosclerotic vascular disease, by ultrasound high-resolution B mode imaging. In analysis 1, it was compared with the predicted mean IMT based on data from existing risk factors (age, male sex, dyslipoproteinaemia, hypertension, diabetes mellitus, smoking status) in 282 non-acromegalic subjects. In analysis 2, the mean IMT in the 21 acromegalic patients was compared with that in 42 non-acromegalic subjects matched for age, sex and the other atherosclerotic risk factors. We also analysed clinical characteristics between the acromegalic patients with and without the atherosclerosis. RESULTS: Mean IMT in 21 acromegalic patients was 0.92 +/- 0.21 (mean +/- SD) mm. It was significantly (P < 0.05) lower than the mean IMT (1.03 +/- 0.12 mm) predicted from their existing risk factors (analysis 1). It was also less than that in 42 non-acromegalic subjects matched for atherosclerotic risk factors (1.07 +/- 0.37 mm; P < 0.05) (analysis 2). Among the acromegalic patients, 10 patients (48%) had increased mean IMT (> or = 1.1 mm) and/or plaque lesions whereas the other 11 had no such atherosclerotic changes. In the patients without the atherosclerotic changes, plasma insulin-like growth factor-I (IGF-I) concentration was significantly (P < 0.01) higher, and the prevalence of hypertension was significantly (P < 0.05) lower than in those with the atherosclerotic changes. CONCLUSIONS: The extent of carotid atherosclerosis in the acromegalic patients was not higher than that in non-acromegalic subjects, considering their atherosclerotic risk factors. Increased concentration of IGF-I might be involved in the lack of susceptibility to atherosclerosis in some acromegalic patients. PMID- 11422115 TI - The improvement of insulin resistance in patients with adrenal incidentaloma by surgical resection. AB - OBJECTIVE: Several recent studies have indicated that patients with adrenal incidentaloma often have disturbed glucose tolerance or/and hypertension. It is unclear whether these metabolic conditions could be caused by adrenal incidentaloma. We investigated the prevalence of disturbed glucose tolerance, hypertension and insulin resistance in the patients with non-functioning adrenal incidentaloma and evaluated the changes of the parameters such as glucose tolerance, blood pressure and insulin sensitivity after adrenalectomy. PATIENTS AND METHODS: Among 15 patients with incidentally discovered adrenal tumours in our department from 1996 to 1999, 4 patients were diagnosed as having pre clinical Cushing's syndrome and the other 11 as having non-functioning tumours based on detailed endocrinological examinations including dexamethasone suppression testing. Four tumours with pre-clinical Cushing's syndrome and 8 tumours out of 11 patients with non-functioning tumours were diagnosed histopathologically as adrenocortical adenomas and the other 3 as of non-adrenal origin including a myelolipoma, an adrenal vascular cyst and an endothelioma. The prevalence of disturbed glucose tolerance was determined with an oral glucose tolerance test, and insulin sensitivity was evaluated by the method of steady state of plasma glucose (SSPG). RESULTS: All 12 patients with adrenocortical adenoma exhibited insulin resistance based on the SSPG (6.9-13.2 mmol/l). Before surgical removal of the tumours, the SSPG titre was relatively higher in the patients with pre-clinical Cushing's syndrome than in those with non-functioning with adrenocortical adenoma (mean value 11.65 vs. 8.99 mmol/l), whereas 2 of the 3 patients with non-adrenocortical tumours did not have insulin resistance. Among the 12 patients with adrenocortical adenoma, 7 (58%) and 9 (75%) patients exhibited hypertension and disturbed glucose tolerance, respectively. After removal of the tumours, SSPG of the patients with adrenocortical adenoma, but not that of the other 3 patients with non-cortical tumours, was significantly decreased compared to pre-adrenalectomy values. There are no significant differences in the changes of SSPG titres between in pre-clinical Cushing's syndrome and in non-functioning adrenocortical adenoma. Systolic blood pressure, but not diastolic blood pressure, was also significantly decreased in the patients with adrenocortical adenoma. CONCLUSION: High prevalences of disturbed glucose tolerance, insulin resistance and hypertension were found among the patients with non-functioning adrenocortical tumours. Adrenocortical adenoma may be one of the risk factors for insulin resistance that is believed to induce disturbed glucose tolerance and/or hypertension. Therefore, it is useful to evaluate insulin resistance for the patients with adrenal incidentalomas since results are likely to be helpful in deciding whether to remove the tumour by surgery. PMID- 11422116 TI - Urate and arteriosclerosis in primary hyperparathyroidism. AB - OBJECTIVE: An increased mortality due to cardiovascular disease has been reported in patients with primary hyperparathyroidism (pHPT). An association between urate and cardiovascular disease has been suggested. Metabolic abnormalities in pHPT may include urate. We therefore evaluated the metabolic arteriosclerotic risk profile in pHPT with special focus on the role of urate. DESIGN: Retrospective analysis of data before and 1 year after surgery for pHPT. PATIENTS: 130 consecutive patients, over the age of 44 years, who underwent surgery for pHPT. MEASUREMENTS: Biochemical variables known to reflect risk of arteriosclerotic disease (AD) and renal function tests including measurement of glomerular filtration rate (GFR) were investigated before and 1 year after surgery. RESULTS: pHPT patients with AD (n = 40) were older and had higher serum levels of urate and triglyceride, and more impaired renal function in comparison with patients without AD. PTH and calcium values did not differ. Multiple logistic regression analysis indicated that urate was an independent risk factor for AD in pHPT (P < 0.01). Three variables were shown to be positively associated with urate; male gender (P < 0.01), fasting blood glucose (P < 0.05) and serum level of triglyceride (P < 0.05). CONCLUSIONS: Urate was found to be an independent risk factor for arteriosclerotic disease in primary hyperparathyroidism. Serum level of urate could (in addition to gender) be associated with a metabolic disorder comprising increased glucose and triglyceride levels. PMID- 11422117 TI - An influence of variation in the aldosterone synthase gene (CYP11B2) on corticosteroid responses to ACTH in normal human subjects. AB - OBJECTIVE: Previous evidence suggests that the efficiency of 11beta-hydroxylase is at least partly heritable and also that it may be mildly impaired in essential hypertension. In both cases, assessment of activity was based on the response of 11-deoxycorticosterone (DOC) and 11-deoxycortisol to ACTH. The gene (CYP11B1) coding for this enzyme is highly homologous with and lies a relatively short distance downstream from the gene coding for aldosterone synthase (CYP11B2) on chromosome 8. Two polymorphisms of CYP11B2 have been described. The first involves a change of -344C to T in a putative steroidogenic factor-1 (SF-1) binding site and the other, the intron conversion, an exchange of intron 2 for that of CYP11B1. These polymorphisms are in linkage dysequilibrium. Their effects on 11beta-hydroxylation were studied. METHODS AND RESULTS: Normal subjects (n = 135) were genotyped and those homozygous for either or both the polymorphisms were given ACTH (250 microg, i.v.). Plasma was sampled before and 30 minutes after administration. Basal concentrations of DOC, corticosterone, 11 deoxycortisol and cortisol and responses of corticosterone and cortisol to ACTH were not affected by genotype. However, the responses of DOC (P = 0.002 and P = 0.001, respectively) and 11-deoxycortisol (P = 0.025 and P = 0.002, respectively) were significantly greater in subjects homozygous for SF-1 T and/or intron conversion than in those homozygous for SF-1 C and/or normal intron. CONCLUSIONS: These results indicate different 11beta-hydroxylase efficiencies. Thus, variation in CYP11B2 appears to affect the product of CYP11B1. The mechanism is unclear. The close proximity of the two genes may lead to competition for transcription factors or specific differences in intron 2 may affect transcription. Alternatively, the polymorphisms may be acting as markers for adjacent functional genetic variations. PMID- 11422118 TI - The influence of growth hormone replacement on heart rate variability in adults with growth hormone deficiency. AB - OBJECTIVE: Growth hormone (GH) deficiency is associated with increased cardiovascular morbidity and mortality. Abnormalities in heart rate variability (HRV), a surrogate marker of cardiac autonomic tone, have also been found in untreated growth hormone deficient (GHD) patients. Similar abnormalities have been found in patients with complications postmyocardial infarction. DESIGN AND MEASUREMENTS: This study was designed to investigate whether GH treatment leads to normalization of cardiac autonomic tone. HRV measurements were obtained from 15 minute resting ECG recordings in 11 untreated adult GHD patients (7 females; mean age 39.2 years), 10 GHD patients (6 females; mean age 46.2 years) reCENving GH replacement (mean duration, 52.7 months) and 12 healthy controls (7 females; mean age 44.6 +/- 2.9 years) who were all of similar age, weight and BMI. The untreated GHD patients were then commenced on GH and HRV measurements repeated after 3 and 6 months of treatment. RESULTS: In accord with our previous findings, HRV analysis using Fast Fourier Transform (FFT) showed a distinct pattern of abnormality in GHD patients compared with controls. Specifically, there was an increase in nHF power (P = 0.04) and a reduction in nLF power (P = 0.04) (representing parasympathetic and sympathetic activity, respectively), a reduction in nVLF power (P = 0.03) and a 50% reduction in LF/HF ratio (P = 0.02) (a measure of sympathovagal balance) in GHD patients when compared with controls. HRV results in patients who have been on long-term GH replacement were indistinguishable from controls. After 3 months GH replacement in the untreated GHD patient group, nVLF power had increased by 28% (P = 0.03) at 3 months and this was sustained at 6 months. However, no significant changes in LF and HF power were seen. CONCLUSIONS: These results suggest that abnormalities of the cardiac autonomic system in GHD patients may be correctable. Longer duration of prospective follow-up will be required to determine at what time point improvements in the other frequency bands occur. PMID- 11422119 TI - A novel mutation (N233K) in the transactivating domain and the N756S mutation in the ligand binding domain of the androgen receptor gene are associated with male infertility. AB - OBJECTIVE: Resistance to androgens has been suggested as a possible cause of male infertility. This hypothesis is based mainly on binding studies in genital skin fibroblasts but the molecular evidence is sparse. DESIGN: Molecular studies of the androgen receptor gene were performed in 10 azoo- or oligozoospermic men, presenting with clinical signs of low androgen activity-poor virilization and high serum LH despite elevated testosterone levels, but without genital malformations. PATIENTS: Ten men with serum LH >10 IU/l and testosterone >30 nmol/l as well as a low sperm concentration < 20 x 106/ml. MEASUREMENTS: Genomic DNA was prepared from peripheral leucocytes and PCR-amplification of the coding region of androgen receptor was performed, followed by direct sequencing. Identified mutations were reconstructed by site-directed mutagenesis and the functional properties of the mutants were analysed, using transient expression in COS-1 cells and subsequent transactivation assays. Hormone binding assays were performed in genital skin fibroblasts from the patients. RESULTS: Two of the 10 men were shown to have a mutation in the androgen receptor gene. Subject 1, who presented with azoospermia, serum testosterone (T) 50 nmol/l and LH 20 IU/l, had a mutation in exon 1, changing amino acid asparagine 233 to lysine (N233K). In fibroblasts cultured from genital skin, the receptor affinity for 5alpha dihydrotestosterone (DHT) was normal as compared to healthy controls, but the receptor-hormone complex was thermolabile at 42 degrees C. Subject 2 exhibited severe oligozoospermia and a similar endocrine pattern (T = 50 nmol/l and LH = 25 IU/l). He had a mutation in exon 5 changing asparagine 756 to serine (N756S). The affinity for DHT in cultured genital fibroblasts from this patient was reduced. Transactivation was abnormal for both mutants, N233K reaching 46% and N756S 38% of wild type activity when stimulated with 10 nmol/l DHT. CONCLUSIONS: Androgen receptor mutations may affect sperm production without resulting in genital malformations. Thus, in infertile men with a clinical presentation of poor androgen activity and an endocrine profile compatible with androgen resistance, mutations in the androgen receptor should be taken into consideration. PMID- 11422120 TI - Undermasculinized genitalia in a boy with an abnormally expanded CAG repeat length in the androgen receptor gene. AB - We report an 11-year-old boy with undermasculinized genitalia and an abnormally expanded CAG repeat length at exon 1 of the androgen receptor (AR) gene. He had microphallus and scrotal hypospadias with chordee, and underwent urethroplasty at 4 years of age. At 11 years of age, a gonadotropin releasing hormone (GnRH) test yielded a relatively high leutinizing hormone (LH) response (0.7-->20.4 IU/l) and a relatively low follicle-stimulating hormone (FSH) response (1.7-->4.8 IU/l), and an human chorionic gonadotropin (hCG) test showed sufficient responses of testosterone (0.7-->23.0 nmol/l) and dihydrotestosterone (0.38-->2.95 nmol/l). The CAG repeat length was 44 for the boy and ranged from 12 to 32 for 100 control males. The DNA sequences of the AR gene were normal for the exons 1-8 and for the splice donor, splice acceptor and branch sites. The markedly expanded CAG repeat length appears to be relevant to the undermasculinized genitalia of this boy, because such an expandsion, which has previously been reported only in spinal and bulbar muscular atrophy, is known to reduce AR function. PMID- 11422121 TI - Ectopic ACTH syndrome caused by pulmonary carcinoid tumourlets. AB - The differential diagnosis of Cushing's syndrome is a major challenge to clinical endocrinologists, especially those infrequent cases referred to as occult ectopic ACTH syndromes. Although bronchial carcinoids are well known to be a cause of Cushing's syndrome due to ectopic ACTH secretion, very few cases of carcinoid tumourlets causing an ACTH ectopic syndrome have been reported, and their origin remains controversial. For some authors, tumourlets and typical carcinoids represent distinct pathological entities, whilst others hold that tumourlets are merely microscopic carcinoid tumours. We report a patient with an aggressive Cushing's syndrome that required bilateral adrenalectomy, diagnosed 22 years before a 3-cm lung nodule became apparent on routine chest X-ray. The biopsy after lung surgery revealed a typical peripheral bronchial carcinoid surrounded by tumourlets. Both tumourlets and carcinoid tumour showed strongly positive ACTH immunostaining. Recently, Arioglu et al. (1998) reported a case of Cushing's syndrome caused by pulmonary carcinoid tumourlets, concluding that this entity should be considered in the differential diagnosis of occult ectopic ACTH syndrome. Furthermore, we consider that the carcinoid tumourlets found in our patient, were the initial source of ACTH, leading to Cushing's syndrome with a rapid onset, and that a loss of cell proliferation control in one of such tumourlets many years later, could have resulted in the development of a typical carcinoid tumour, reinforcing the theory of a common origin of these lesions. PMID- 11422122 TI - Re: hyponatremia. PMID- 11422123 TI - IL-10: a key regulator of allergic disease. PMID- 11422124 TI - On allergens and asthma (again): does exposure to allergens in homes exacerbate asthma? PMID- 11422125 TI - Rhinitis and asthma: a continuum of disease? PMID- 11422126 TI - Pathogenesis of systemic lupus erythematosus (SLE). PMID- 11422127 TI - Management of systemic lupus erythematosus (SLE). PMID- 11422128 TI - Inhibition of IgE-induced activation of human mast cells by IL-10. AB - BACKGROUND: IL-10 exhibits anti-inflammatory effects on activated rodent mast cells (MC) in vitro and inhibits allergen-induced airway inflammation in vivo in murine models. The effects of IL-10 on the allergic activation of human MC are presently unknown. OBJECTIVE: In light of the well-known heterogeneity of mast cell reactivity between animal species, one cannot readily predict the response of human MC to IL-10. Moreover, the impact of IL-10 on MC-derived proinflammatory mediators is still unknown. Thus, the objective of this study was to investigate the effects of IL-10 on the release of inflammatory mediators by IgE/anti-IgE challenged human cord blood-derived mast cells (CBMC), used as an in vitro model of MC phenotypically similar to human lung MC. MATERIALS AND METHODS: Highly purified human MC were obtained by a first step of long-term culture of cord blood mononuclear cells in the presence of human recombinant stem cell factor (rhSCF) and of human recombinant IL-6 (rhIL-6), followed by a second step of purification by depletion of contaminating cells with an immunomagnetic METHOD: The cells were treated with human IgE, then challenged with anti-human IgE, in the presence or the absence of recombinant rhIL-10 used at various concentrations. Histamine, tumour necrosis factor-alpha (TNF-alpha), IL-5 and IL 8 were measured in the various supernatants collected at different times after the beginning of the challenge. RESULTS: IL-10 inhibited the release of TNF-alpha and of IL-8, but not of IL-5, by activated CBMC. Interestingly, IL-10 also inhibited the release of histamine by activated CBMC, contrasting with data reported for rodent MC. CONCLUSIONS: These findings suggest that IL-10 might have anti-inflammatory effects on IgE/anti-IgE-challenged human MC by inhibiting their release of TNF-alpha, IL-8 and histamine. These data provide new insights into the control of human mast cell activation and might lead to a better knowledge of the cellular mechanisms controlling allergic reactions. PMID- 11422129 TI - Der p 1 and Der p 2 induce less severe late asthmatic responses than native Dermatophagoides pteronyssinus extract after a similar early asthmatic response. AB - BACKGROUND: The models for exposure to house dust in research and clinical practice are selected with respect to their role in IgE-mediated immediate hypersensitivity. The use of isolated major allergens instead of complex allergen extracts is becoming increasingly popular as it offers some important advantages for quantitative measures in diagnosis and research. OBJECTIVE: To compare house dust mite extract and isolated mite major allergens with respect to their ability to induce early and late asthmatic responses and bronchial hyperreactivity. METHODS: Bronchial responses to house dust mite (HDM, Dermatophagoides pteronyssinus) extract and isolated major allergens from HDM (Der p 1 and Der p 2) were compared in a double-blind, randomized, cross-over study in 20 patients with mild to moderate asthma who were allergic to HDM. Allergen was titrated to a standardized early asthmatic response. Bronchial hyper-responsiveness to histamine (PC20histamine) was determined before and after allergen inhalation to assess allergen-induced bronchial hyper-responsiveness and IL-5 was measured in serum. In addition, the allergens were applied in intracutaneous skin tests and activation of basophil leucocytes and proliferation of peripheral blood mononuclear cells was tested in vitro. RESULTS: After a similar early asthmatic response (mean Deltaforced expiratory volume in 1 s (FEV1),max -29.4 (SD 7.2) vs. -33.1 (8.6) %; mean difference 3.6 (95% CI -0.9 to 8.2) %), the late asthmatic response (mean DeltaFEV1,max -45.9 (21.9) vs. -32.7 (22.3) %; mean difference 13.2 (3.8-22.3) %), the degree of allergen-induced bronchial hyper-responsiveness (mean DeltaPC20histamine, 1.8 (1.0) vs. 1.2 (0.9) doubling dose; mean difference 0.6 (0.2-1.1) doubling dose) and serum IL-5 at 6 h were found to be significantly higher after bronchial challenge with HDM extract than after challenge with an isolated HDM major allergen. Likewise, there was an increased late skin reaction with HDM compared with isolated major allergen after a similar early skin reaction. CONCLUSION: Constituents of HDM extract, other than Der p 1 or Der p 2, with no significant influence on the IgE-mediated early asthmatic response contribute significantly to the allergen-induced late asthmatic response and bronchial hyper-reactivity. PMID- 11422130 TI - Low domestic exposure to house dust mite allergens (Der p 1) is associated with a reduced non-specific bronchial hyper-responsiveness in mite-sensitized asthmatic subjects under optimal drug treatment. AB - BACKGROUND: Airway inflammation in asthma causes symptoms, airflow limitation and bronchial hyper-responsiveness. The strategy of asthma management is to reduce airway inflammation by drug treatment and avoidance of triggers, including allergens. OBJECTIVE: We determined the effect of exposure to house dust mite (HDM) allergens on bronchial responsiveness in asthmatics sensitive to mites while under optimal drug treatment. METHODS: We studied 71 mild to moderate HDM sensitive asthmatics. Drug treatment sufficient to keep asthma under control was administered to each patient for 1 year. Subjects were divided into two groups, according to the amount of Der p 1 in their bedrooms measured after standard HDM reduction measures: low Der p 1 exposure (0.64 +/- 0.5 microg/g dust) (Group 1, n = 34) and high Der p 1 exposure (12.5 +/- 11.4 microg/g) (Group 2, n = 37). Bronchial responsiveness to methacholine (PD20FEV1) was determined at the beginning and end of the study. RESULTS: In Group 1, PD20FEV1 increased 2.15-fold at the end of the study from 57 to 123 microg (P < 0.05), whereas in Group 2 no significant changes were observed. The subjects in Group 2 tended to increase the use of inhaled steroids and bronchodilators in the autumn months compared with subjects in Group 1, but the difference was not significant. CONCLUSION: This long-term study shows that exposure to lower levels of mite allergens in the bedroom is associated with a decrease of bronchial hyper-responsiveness in sensitized asthmatic subjects under optimal drug treatment. PMID- 11422131 TI - Inhaled corticosteroids decrease vascularity of the bronchial mucosa in patients with asthma. AB - BACKGROUND: Increased vascularity in airway mucosa is a distinctive feature of airway remodelling in asthma. While corticosteroids have proved most effective in modifying airway inflammation, the effect of inhaled corticosteroids on increased airway mucosal vascularity in asthmatics has been little studied. OBJECTIVE: We examined the effect of inhaled corticosteroid on airway vascularity in bronchial biopsy specimens taken from asthmatic patients. SUBJECTS AND METHODS: We studied bronchial biopsies from 28 asthmatic patients before and after treatment with inhaled beclomethasone dipropionate (BDP) 800 microg/daily, or placebo, for 6 months in a double-blind manner. Biopsy specimens were evaluated for number of vessels and percentage of area occupied by vessels, using computerized image analysis after staining for type IV collagen in vessel walls. Specimens were also examined for extent of collagen III in the subepithelial basement membrane. In addition, we compared asthmatic specimens with biopsy specimens taken from non asthmatic control subjects. RESULTS: There was a significant increase in number of vessels (P < 0.01) and percent vascularity (P < 0.001) in the submucosa of asthmatic patients compared with control subjects. After 6 months of treatment, we observed significant improvements in forced expiratory volume in 1 s (FEV1), FEV1% and airway responsiveness (P < 0.05, each) in the BDP treatment group compared with the placebo group. This was accompanied by significant decreases in both vessel number and percent vascularity in the airways of BDP-treated patients (P < 0.05, each). We also observed a significant correlation between change in percent vascularity and change in collagen III thickness in the BDP-treated patients (rs = 0.90, P < 0.001). Furthermore, the change in percent vascularity was inversely correlated with both FEV1 (rs = -0.49, P < 0.05) and airway responsiveness (rs = -0.36, P < 0.05). CONCLUSION: These findings suggest that inhaled corticosteroid treatment of asthma reduced airway wall vascularity during airway remodelling. PMID- 11422132 TI - IFN-gamma but not IL-4 T cells of the asthmatic bronchial wall show increased incidence of apoptosis. AB - BACKGROUND: Previous observations have established that IFN-gamma production is depressed in CD4+ T cells from atopic asthmatics compared with non-asthmatics. OBJECTIVE: The aim of this study was to determine if decreased IFN-gamma production could be due to a dissociation between levels of apoptosis within the T cell subsets of the asthmatic bronchial wall. METHODS: Twenty asthmatics (10 atopic and 10 non-atopic) and eight non-atopic non-asthmatics underwent bronchoscopy. Cryostat sections of these biopsies were investigated using immunohistological techniques to determine the relative number of CD4/FAS+ and CD4/Bcl-2+ cells. Detection of IFN-gamma+ and IL-4+ was combined with TUNEL staining to determine the proportions of the Th1 and Th2 cells undergoing apoptosis. RESULTS: Experiments revealed raised proportions of activated CD4+ T cells as assessed by expression of HLA-DR and CD25+ expression in the asthmatic samples. Expression of Bcl-2 by the CD4+ cell population was significantly reduced in the asthmatic compared with the control group (P = 0.002). There was no significant difference in the expression of CD4+ Fas-ligand or the number of CD4+ undergoing apoptosis in the asthmatic and non-asthmatic groups. However, the IFN-gamma+ (P = 0.04) but not IL-4+ T cells in the asthmatic biopsies had significantly higher proportions of apoptotic cells compared with the control group. CONCLUSION: The evidence supports the hypothesis that Th1/Th2 imbalance in asthmatic inflammation may be a result of premature apoptosis within the Th1 subset. PMID- 11422133 TI - Wasp venom immunotherapy induces a shift from IL-4-producing towards interferon gamma-producing CD4+ and CD8+ T lymphocytes. AB - BACKGROUND: Venom immunotherapy (VIT) has proven to be very effective in hymenoptera venom anaphylaxis. However, the underlying immunoregulatory mechanisms of venom immunotherapy remain poorly understood. Recent studies measured the total amount of cytokine in culture supernatans, suggesting a shift in cytokine production from Th2 to a Th1 cytokine profile during VIT. We wanted to examine the contribution of specific T lymphocyte subpopulations, which is impossible using an extracellular method to determine cytokines in supernatants. OBJECTIVE: The present study was designed to evaluate the effect of VIT on the percentages of type 1 (IL-2, interferon-gamma (IFN-gamma)) and type 2 (IL-4) CD4+ and CD8+ T lymphocytes of patients with wasp venom anaphylaxis during immunotherapy. METHODS: Peripheral blood mononuclear cells of 20 individuals with a history of wasp sting anaphylaxis and a positive serum wasp venom specific IgE were isolated and in vitro stimulated with phorbol-12-myristate-13-acetate and ionomycin before VIT, at the end of a 5-day semirush VIT and at 6 months during VIT. Three-colour flow cytometric analysis was used for intracellular cytokine (IL-2, IFN-gamma, IL-4) detection in CD4+ (CD3+CD8-) T lymphocytes and CD8+ (CD3+CD8+) T lymphocytes. RESULTS: At the end of a 5-day semirush VIT, there was a significant decrease in percentage of IL-4-producing CD4+ and CD8+ T cells, compared with cytokine-producing cells before VIT (P = 0.0002 and 0.004). After 6 months of VIT, patients showed an increased number of IL-2-producing stimulated CD4+ and CD8+ lymphocytes compared with values before VIT (P = 0.002 and P = 0.0003). A higher amount of IFN-gamma-producing stimulated CD4+ and CD8+ cells was found after 6 months of VIT (P = 0.001 and P = 0.0006). There was no correlation between cytokine-producing cells and specific IgE for wasp. CONCLUSION: Venom immunotherapy induced a shift from IL-4-producing towards IFN gamma-producing CD4+ as well as CD8+ T lymphocytes. PMID- 11422134 TI - Diet and chronic obstructive pulmonary disease: independent beneficial effects of fruits, whole grains, and alcohol (the MORGEN study). AB - BACKGROUND: In recent years antioxidants, foods rich in antioxidants (e.g. fruits, vegetables) and fish have been suggested to protect against chronic obstructive pulmonary disease (COPD). There are also indications for a protective effect of whole grain intake and of consuming moderate amounts of alcohol. It is, however, not clear whether the effects of the different dietary factors on COPD are independent of each other and if so, whether their effects are additive. OBJECTIVE: To gain more insight into the potential protective effect of diet on COPD, we studied fruit, vegetable, fish, alcohol and whole grain consumption simultaneously in relation to pulmonary function and COPD symptoms. METHODS: Analysed were cross-sectional data collected in 13 651 men and women aged 20-59 years participating between 1994 and 1997 in the MORGEN study (monitoring project on risk factors and health in The Netherlands). Regression models were adjusted for age, gender, height (for pulmonary function only), smoking, BMI and energy intake. RESULTS: Fruit and whole grain intake showed independent beneficial associations with COPD (P-trend < 0.001). Furthermore, in subjects with low alcohol consumption (1-30 g/day) the forced expiratory volume in 1 s (FEV1) was higher and the prevalence of COPD symptoms lower than in non-drinkers (P < 0.001). The effects of a favourable intake of fruits (> 180 g/day), whole grains (> 45 g/day) and alcohol (1-30 g/day) were largely additive. In the 2998 subjects with a favourable intake of the three foods, the FEV1 was 139 mL higher and the prevalence of COPD symptoms lower (odds ratio (OR) = 0.44) than in subjects (n = 1406) with unfavourable intakes of fruits, whole grains and alcohol (P < 0.001). A similar effect was observed in those who had never smoked. Fish and vegetable intake did not show independent beneficial associations with COPD. CONCLUSIONS: Our results suggest independent beneficial effects of fruits, whole grains and alcohol on COPD that are largely additive and cannot be explained by smoking habits. PMID- 11422135 TI - A simple diagnostic index for asthma. AB - BACKGROUND: Asthma is becoming increasingly prevalent and a number of research groups are investigating its genetic and environmental basis. OBJECTIVE: To produce a brief screening tool suitable for determining phenotype in asthma research. METHODS: The scores from eight questions on symptoms and history were obtained from 678 adults and 244 children from high asthma-incidence caucasian families. An independent physician diagnosis was also obtained with the use of a modified NHLBI-CSGA questionnaire and pulmonary function test. Stepwise logistic regression was applied to determine which of the eight questions had greatest predictive value for asthma, and the quality of the resultant models was evaluated using an independent set of 643 adults and 239 children. RESULTS: For adults, the most parsimonious model used responses from three of the eight questions. It had sensitivity and specificity of 0.94 and 0.96, respectively. For children, responses to two questions gave a model with sensitivity and specificity of 0.97. For both age groups, negative predictive values were above 0.87. Positive predictive values were 0.58 and 0.78 for adults and children respectively. The latter emphasize the need for conformation, by physician, of "affected" calls made by this initial screen. CONCLUSION: The brief questionnaires described are potentially useful in a research setting, as a preliminary screening mechanism of low cost. Their use will reduce the numbers of subjects that must undergo detailed phenotyping. PMID- 11422136 TI - Cloning and expression of Aspergillus fumigatus allergen Asp f 16 mediating both humoral and cell-mediated immunity in allergic bronchopulmonary aspergillosis (ABPA). AB - BACKGROUND: Aspergillus fumigatus, a ubiquitous fungus, is responsible for a number of lung disorders in atopic and non-atopic individuals. Standardized, pure, and relevant allergens are desirable for reliable immunodiagnosis of the disease and to understand the structural and functional properties of these allergens and the role they play in causing ABPA. OBJECTIVE: Molecular cloning and characterization of a relevant allergen from A. fumigatus cDNA library. MATERIALS AND METHODS: A cDNA library was constructed from 96 h old mycelium of A. fumigatus using lambda ZAP expression vector. A novel gene encoding an A. fumigatus allergen was identified by screening the library with sera from ABPA patients. The gene was cloned and the allergen over-expressed in Escherichia coli. This recombinant allergen, Asp f 16, was evaluated in ELISA and Western blots using sera from patients and normal subjects and peripheral blood mononuclear cells (PBMC) for antigen-induced stimulation. RESULTS: Seventy percent of the patients with ABPA demonstrated high levels of serum IgE antibodies to Asp f 16, a 43-kDa protein, whereas patients with allergic asthma, Aspergillus skin test-positive asthmatics without clinical evidence of ABPA, and normal controls failed to show Asp f 16-specific IgE binding by ELISA. The deduced amino acid sequences of Asp f 16 showed extensive sequence homology to 30.6-kDa Asp f 9 at the N-terminal region of the protein. PBMC from the majority of patients with ABPA exhibited significant proliferation with the recombinant Asp f 16 allergen. CONCLUSION: Specific humoral and cell-mediated immune responses of Af-sensitized patients against Asp f 16 suggest its usefulness in the immunodiagnosis of hypersensitivity diseases due to Af and understanding the pathophysiology of ABPA. PMID- 11422137 TI - Identification of mutations in the genes for the pollen allergens of eastern red cedar (Juniperus virginiana). AB - BACKGROUND: Cedar pollens are important causes of seasonal allergic disease in diverse geographical areas. However, pollens from different families and species vary in their propensity to induce allergic responses. OBJECTIVE: To compare the structure of potential allergens from eastern red cedar (Juniperus virginiana) pollen with those of the highly allergenic cedar (mountain cedar, J. ashei) pollens. MATERIALS AND METHODS: The cDNAs for potential pollen allergens, Jun v 1 and Jun v 3, were amplified by reverse transcriptase-polymerase chain reaction, cloned and sequenced. Expression of the native proteins in pollen was characterized by SDS-PAGE and immunoblotting. RESULTS: The cDNA sequence for one potential major allergen, Jun v 1, was highly homologous to those of the other cedar pollens. The second potential allergen, Jun v 3, was also highly homologous to its counterpart in mountain cedar, but a stop codon in the mRNA would result in a protein of only 91 amino acids, which would lack potential N-glycosylation sites and the IgE binding epitopes of the 199 amino acid homologue from mountain cedar pollen, Jun a 3. IgE from the sera of patients with hypersensitivity to cedar pollen bound to eastern red cedar proteins of four different sizes. N terminal amino acid sequence analysis indicated that two of these proteins (43 and 30 kDa) were either isoforms or processed Jun v 1. No Jun v 3 protein was detected. The N-terminal sequence of an additional 145-kDa allergen, termed Jun v 4, was not homologous to any previously described allergens. CONCLUSION: These findings suggest that mutations in the genes or post-translational modifications of two potentially allergenic proteins might help to explain why the pollen of eastern red cedar is reported to be less allergenic than those of other members of Cupressaceae and Taxodiaceae families. PMID- 11422138 TI - IgE-mediated occupational asthma induced by herbal medicine, Banha (Pinellia ternata). AB - There have been few reported cases of occupational asthma induced by Pinellia ternata (Banha), and the mechanism responsible for this type of asthma is still undetermined. We report a case of Banha-induced occupational asthma with IgE mediated mechanism. The patient had positive skin responses to Banha extract and Banha-specific bronchial challenge elicited an early asthmatic response. The serum-specific IgE binding to Banha extract was detectable and completely inhibited with the additions of 0.1 microg/mL of Banha extract on ELISA inhibition. Seven IgE binding components to Banha extract (6.5, 22, 24, 32, 34, and 48 kDa) were detected using SDS-PAGE and immunoblot analysis. In conclusion, the results of this study suggest that P. ternata (Banha)-derived allergens are able to cause IgE-mediated bronchoconstriction in exposed workers. PMID- 11422139 TI - Eosinophilic rhinitis accompanies the development of lower airway inflammation and hyper-reactivity in sensitized mice exposed to aerosolized allergen. AB - BACKGROUND: Allergic rhinitis is a risk factor for the development of asthma. About 80% of asthmatic patients also have rhinitis. However, the pattern of induction of allergic rhinitis and asthma remains unclear. OBJECTIVE: The purpose of this study was to investigate the development of upper airway inflammation in mice during the development of an asthma-like disease and after an acute allergen provocation. METHODS: BALB-c mice were sensitized intraperitoneally (i.p) to ovalbumin (OA, days 1-13) and were challenged with aerosols of either OA or saline on 8 consecutive days (days 33-40). In a second experiment, chronic exposure for 8 days was followed by 10 days of rest and then an acute nebulized allergen provocation was performed (day 50). Inflammatory parameters were investigated at different time-points. RESULTS: Upper and lower eosinophilic airway inflammation were simultaneously induced in the course of repeated inhalations of nebulized OA, as shown by analyses of nasal and broncho-alveolar lavage fluids and histological sections of the nose and bronchi. Mice that developed bronchial hyper-responsiveness also had increased thickness of the nasal mucosa on magnetic resonance image (MRI) scans. When chronic exposure was followed by acute allergen provocation, the latter caused a systemic increase in IL-5 levels, with a concomitant rise in blood and airway eosinophils, primarily in the nose. CONCLUSIONS: Simultaneous induction of eosinophilic inflammation in the nose and lungs was found in a mouse model of respiratory allergy. These findings support the viewpoint that upper and lower airway disease represent a continuum of inflammation involving one common airway and provide evidence for the concept of global airway inflammation after inhalation of allergen. PMID- 11422140 TI - Cytokine mRNA expression in patients with mild allergic asthma following low dose or cumulative dose allergen provocation. AB - BACKGROUND: Allergen provocation is a very useful way to study the inflammatory response in asthmatic patients. Although cumulative dose regimens are most often applied, another provocation model with repeated inhalations of low doses of allergens has recently come into use. OBJECTIVE: We were interested to compare these two allergen provocation models. To evaluate the inflammation induced by either model, we examined the mRNA expression of several cytokines that are implicated in the orchestration of the inflammatory response observed in asthma. METHODS: Interleukin (IL)-4, IL-5, IL-13 and interferon (IFN)-gamma mRNA expression was analysed in bronchoalveolar lavage (BAL) cells and peripheral blood (PB) CD4+ and PB CD8+ T cells following any of the two provocation regimens. IL-4 and IFN-gamma mRNA expression was analysed by a competitive reverse transcriptase-polymerase chain reaction (RT-PCR) method, while IL-5 and IL-13 were analysed semiquantitatively, before and after allergen provocation with either model. RESULTS: After low dose provocations none of the cell populations studied showed a clear change in the pattern of IL-4 or IFN-gamma gene expression. In contrast, after cumulative dose provocation there was a clear tendency towards an increased IL-4 mRNA expression in BAL cells, correlating with a significant increase in IL-4 mRNA in PB CD4+ as well as in CD8+ T cells (P = 0.005 and P = 0.04, respectively). Regardless of the allergen provocation method used, in PB IL-4 mRNA was preferentially expressed by CD4+ cells while IFN-gamma was expressed more by CD8+ cells. IL-5 transcripts increased after low dose provocations in PB CD4+ T cells in six of eight patients, while after cumulative dose provocation IL-5 mRNA increased in BAL cells in six out of nine patients but decreased especially in PB CD8+ T cells in six out of eleven patients, suggesting an accumulation of IL-5 expressing cells to the lungs. CONCLUSION: Thus, the cumulative dose provocation regimen can induce a more pronounced Th2-like immune response in asthmatic patients than the low dose provocation model. PMID- 11422141 TI - Housing characteristics and mite allergen levels: to humidity and beyond. PMID- 11422142 TI - Does leukotriene modulation of eosinophil function explain the therapeutic effectiveness of receptor antagonists in some patients with asthma? PMID- 11422143 TI - Mutual activation of pulmonary fibroblasts and eosinophils, and modulation by drugs in relation to asthma. PMID- 11422144 TI - Integrin VLA-5: modulator and activator of mast cells. PMID- 11422145 TI - Management of seasonal allergic conjunctivitis (SAC): current therapeutic strategies. PMID- 11422146 TI - The importance of housing characteristics in determining Der p 1 levels in carpets in New Zealand homes. AB - BACKGROUND: A previous study of homes in Wellington, New Zealand showed that having carpets on floors was the most important determinant of floor Der p 1 levels, but there was much unexplained variability between houses in carpet levels. OBJECTIVE: To determine to what extent housing characteristics might explain this variability in Der p 1 levels between houses. METHODS: We returned to a selection of houses with carpets and sampled living room dust from 1 square metre for 1 min and from the whole floor at 5 m(2) per min. Der p 1 levels were estimated by double monoclonal antibody ELISA and are expressed as geometric mean microg/g and microg/m(2) (95% confidence intervals). Questionnaires were used to collect information on housing characteristics. RESULTS: Der p 1 levels were significantly higher in the 1 square metre sample (40.0, 31.9-50.2 microg/g; 53.4, 41.4-68.9 microg/m(2)) than in the whole room (25.8, 21.3-31.1 microg/g; 5.3, 3.8-7.4 microg/m(2)). However, results from the different sampling methods were correlated (r = 0.51, P = 0.001 for microg/g and r = 0.58, P = 0.001 for microg/m(2)). After controlling for possible confounders, houses with insulation or a room or garage below the living room had approximately half the Der p 1 concentration (P = 0.05 for both samples) and the amount of Der p 1 per m(2) (P = 0.004 for the 1 square meter sample, P = 0.06 for the whole room sample) than houses without these features. Having more than two children was associated with higher levels of Der p 1 in 1 square meter, significant (P = 0.05) for microg/m(2). Carpet underlay less than 8 mm thick was associated with an almost 3 fold increase in microg/m(2) Der p 1 (P = 0.03) and a 1.6-fold increase in microg/g Der p 1 (P = 0.08) in the whole room sample, when compared with thicker carpet underlays. CONCLUSION: The presence of insulation is the single most important housing characteristic explaining the between-house variability in Der p 1 levels on carpeted living room floors. PMID- 11422147 TI - Effect of a specific cysteinyl leukotriene-receptor 1-antagonist (montelukast) on the transmigration of eosinophils across human umbilical vein endothelial cells. AB - BACKGROUND: Leukotrienes have been implicated in the selective infiltration of eosinophils into the bronchial mucosa in asthma. OBJECTIVE: We studied whether eosinophil transmigration through cultured human umbilical vein endothelial cells (HUVECs) can be blocked by a specific cysteinyl LT1-receptor-antagonist. METHODS: Unstimulated and stimulated eosinophils from patients with asthma and normal controls were subjected to confluent human umbilical vein endothelial cell (HUVEC) monolayers separating the upper and lower chamber of Transwell culture plates. Unstimulated eosinophils or cells pre-incubated in the presence of the eosinophil activating cytokines GM-CSF or IL-13 were placed in the upper chambers while PAF, a potent chemoattractant factor for eosinophils, was added to the lower chamber. Migration of eosinophils was quantified by a beta-glucuronidase assay. RESULTS: The assumption that eosinophils express CysLT1 (cysteinyl leukotriene 1)-receptors was based on our demonstration of mRNA-expression for the CysLT-1-receptor by polymerase chain reaction on purified eosinophils. The chemotactic response to PAF was significantly reduced when eosinophils were pre incubated with montelukast for 15 min. When eosinophils were pre-incubated with GM-CSF and/or IL-13, the migratory response to PAF was also significantly reduced by montelukast. CONCLUSION: From these data we conclude that the specific cysteinyl LT1-receptor antagonist montelukast can inhibit PAF-induced eosinophil transmigration through cultured HUVEC monolayers. PMID- 11422148 TI - Long-term asthma control with oral montelukast and inhaled beclomethasone for adults and children 6 years and older. AB - BACKGROUND: Leukotriene receptor antagonists have demonstrated clinical benefits in chronic asthma studies of up to 3 months in duration. The effects of these agents over extended periods of time have not been reported. OBJECTIVE: To describe the long-term effect of oral montelukast, a potent and specific cysteinyl leukotriene receptor antagonist, compared with inhaled corticosteroids in both adult and paediatric patients with chronic asthma. METHODS: Male and female patients with chronic, stable asthma (adults aged 15-85 years, children aged 6-14 years), who had completed double-blind, placebo-controlled clinical studies, participated in three extension studies with oral montelukast taken once daily (10 mg tablet for adults, 5 mg chewable tablet for paediatric patients) or inhaled corticosteroids (beclomethasone 200 microg twice daily for adults, beclomethasone 100 microg or equivalent three times daily for children). A double blind adult extension study was 37 weeks in duration; open-label adult extension studies were 156 (adults) and 112 (paediatric) weeks in duration. A total of 436, 374, and 245 patients entered these extension studies, respectively. RESULTS: Treatment with both montelukast and inhaled corticosteroids resulted in improvement in multiple parameters of asthma control. Improvements in daytime symptom scores were generally comparable among treatment groups. No tachyphylaxis to the effects of montelukast was evident. In the adult open-label study, however, the effect of beclomethasone on mean forced expiratory volume in 1 second (FEV1) gradually decreased from start of the study to the end of the follow-up treatment period. CONCLUSION: Both montelukast and inhaled corticosteroids were effective in controlling mild to moderate chronic asthma; the relative effectiveness of montelukast and beclomethasone were similar in open label conditions. The hypothesis, that clinical practice conditions (e.g., adherence) may have a significant impact on the effectiveness of these therapies, should be tested in future clinical trials. PMID- 11422149 TI - Comparison of once daily fluticasone propionate aqueous nasal spray with once daily budesonide reservoir powder device in patients with perennial rhinitis. AB - BACKGROUND: Previous studies comparing the corticosteroids fluticasone propionate (FP) and budesonide (BUD) in both perennial and seasonal rhinitis have shown no consistent difference between treatments. However, the therapeutic outcomes may have been influenced by study design. OBJECTIVE: To compare the effect of FP aqueous nasal spray (ANS; 200 microg/day) with BUD reservoir powder device (RPD; 200 microg/day) on rhinitis symptoms, productivity loss and device preference in patients with perennial rhinitis. METHODS: After a 2-week run-in period, 440 patients were randomized to receive either FPANS, BUD RPD or matched placebo (ANS or RPD) for 8 weeks, followed by an open-label 4-week follow-up treatment with FPANS. Patients completed diary card visual analogue scores for nasal symptoms, and questionnaires on satisfaction with the treatment and preferred choice of device. RESULTS: During weeks 1-4, the visual analogue total nasal symptom scores (VATNS) in the FPANS group were significantly lower than scores in the BUD RPD group (mean difference = -17.8; 95% CI = -34.4, -1.3; P = 0.036). FPANS also significantly reduced the VATNS compared with the ANS placebo at all time-points assessed (P < or = 0.005). BUD RPD did not significantly differ from the RPD placebo at weeks 5-8 (P = 0.167), or the ANS placebo at any time-point (P < or = 0.151). Over the 8-week treatment period FPANS was significantly more effective than BUD RPD at reducing sneezing (mean difference = -4.4; 95% CI = -8.6, -0.3; P = 0.036) and nasal itching (mean difference = -5.3; 95% CI = -9.9, -0.8; P = 0.022), and was significantly superior to the ANS placebo for all symptoms assessed at weeks 1-4 and 1-8 (P < 0.016). At the same time-points BUD RPD was no better at alleviating nasal itching than the RPD placebo (P < or = 0.306), and compared with the ANS placebo, significantly reduced only one symptom; nasal blockage (P < or = 0.016). After 8 weeks of treatment, patients preferred the ANS device to the RPD (P < 0.001), and at 12 weeks a significantly greater number of patients were satisfied with FPANS treatment compared with BUD RPD (P = 0.0019) or the respective placebos (P = 0.0001). CONCLUSION: FPANS and BUD RPD are effective therapies with a good safety profile for the treatment of perennial rhinitis but, in this direct placebo-controlled comparison, FPANS was more efficacious than BUD RPD, and the patients preferred the ANS device to the RPD. PMID- 11422150 TI - Evidence for an inflammatory pathophysiology in idiopathic rhinitis. AB - BACKGROUND: The pathophysiology of idiopathic rhinitis is unknown but the disease is classified as being non-allergic on the basis of negative serum IgE radioallergosorbent assay (RAST) and skin prick tests. In contrast, allergic rhinitis has a Th2 type inflammatory pathology mediated by IgE and mast cells. OBJECTIVE: To test the null hypothesis that there would be no difference in the cellular infiltrate for key Th2-associated inflammatory cells between allergic and idiopathic rhinitis. METHODS: We applied strict selection criteria in the recruitment of allergic and idiopathic rhinitis cases. In contrast to previous studies which used cytology or small biopsies, we studied all layers of the mucosa by using whole, full-thickness nasal turbinate specimens with an average length of 2.5 cm. Immunohistochemistry and in situ hybridization techniques were used to compare the distribution and cell populations of mast cells, IgE positive (IgE+) cells, eosinophils and plasma cells in perennial allergic (n = 11) and idiopathic (n = 17) rhinitis, and control nasal mucosal tissue (n = 9). RESULTS: Mast cells and IgE+ cells were significantly increased within the epithelium of allergic and idiopathic mucosa compared to normal mucosa (P < 0.05). More IgE+ cells were present in the allergic group compared to the idiopathic group with the majority of IgE+ cells being mast cells. Both rhinitic groups showed increased eosinophilia localized to the superficial submucosa compared to normal mucosa (P < 0.05). More plasma cells were present in the allergic rhinitic tissue. CONCLUSION: Idiopathic and allergic rhinitic mucosa show similarities in their inflammatory infiltrate suggesting that both groups share a highly localized Th2, IgE-mediated cellular immunopathology. PMID- 11422151 TI - Influenza virus A stimulates expression of eotaxin by nasal epithelial cells. AB - BACKGROUND: Respiratory virus is one of the most common causes of airway inflammation, but its pathogenic mechanisms are not well understood. Eotaxin is a potent eosinophil chemoattractant and is a selective agonist for C-C chemokine receptor 3 (CCR3). Although it has recently been demonstrated that epithelial cells express eotaxin, both in vivo and in vitro, there are few data concerning the expression in viral infection. OBJECTS: We hypothesized that eotaxin may play an important role in attracting inflammatory cells into the airway after viral infection and analysed whether viral infection induces eotaxin in nasal epithelial cells in vitro. METHODS: Nasal epithelial cells obtained from polypectomy for nasal polyp were infected with influenza virus A (subtype H3N2). The cells and supernatants were collected 8, 24 and 48 h after infection. Eotaxin mRNA was analysed by RT-PCR. Eotaxin concentration in the supernatants was analysed by enzyme-linked immunosorbent assay. We also examined a blocking assay to analyse the intervention of pro-inflammatory cytokines, TNF-alpha and IL-1beta in eotaxin production induced by influenza virus. RESULTS: The results showed that eotaxin was expressed constitutively in uninfected cells, but was up regulated for both mRNA and protein levels in infected cells. Blocking experiments using anti-TNF-alpha and anti-IL-1beta antibodies showed no effects of these agents on the level of eotaxin. In addition, UV-inactivated virus did not enhance the expression of eotaxin. CONCLUSIONS: These results suggest that influenza virus A infection in nasal epithelial cells stimulates the expression of eotaxin, and may play an important role in the pathogenesis of airway inflammation by inducing eotaxin. PMID- 11422152 TI - Increased nitric oxide production in nasal epithelial cells from allergic patients--RT-PCR analysis and direct imaging by a fluorescence indicator: DAF-2 DA. AB - BACKGROUND: Nitric oxide (NO) is believed to participate in the regulation of airway clearance and non-specific cellular immunity. Recent studies have suggested that airway epithelial cells of allergic and non-allergic individuals may differ in their ability to produce this molecule. OBJECTIVE: The aim of this study was to detect the difference in NO production in human nasal epithelial cells between normal subjects and patients with perennial allergic rhinitis (AR), and to assess the relationship between the expression of nitric oxide synthase (NOS) isoforms and the severity of the disease. METHODS: Nasal epithelial cells were obtained from the inferior turbinate. The expression of mRNAs encoding constitutive endothelial NOS (eNOS) and inducible NOS (iNOS) was studied by reverse transcription-polymerase chain reaction (RT-PCR). Direct NO production in living cells was visualized and quantified by a fluorescent indicator, DAF-2 DA. RESULTS: RT-PCR analysis demonstrated that AR patients with a RAST score of 5 or 6 showed significant increases in the levels of iNOS mRNA and slight reductions in those of eNOS mRNA. Patients with a RAST score of 2-4 also revealed the same tendency however, the difference was not significant. DAF-2 DA imaging demonstrated that epithelial cells, especially the ciliated cells, produced a larger amount of NO than non-epithelial inflammatory cells. Preincubation with L NAME resulted in an approximate 40% decrease in both groups. CONCLUSION: These results directly indicate that nasal epithelial cells of AR patients overall produce higher levels of NO through the concomitant expression of different NOS isoforms. Continuous NO production by the epithelial cells in normal subjects further support the hypothesis that NO derived from epithelium may play dual roles in the regulation of nasal airway clearance and in the host defense. In addition, the use of DAF-2 DA provides a reliable method to visualize and quantify the direct NO production of living cells. PMID- 11422153 TI - IgE-dependent release of myeloperoxidase by neutrophils from allergic patients. AB - BACKGROUND: The three forms of IgE receptor: the heterotrimeric high-affinity receptor for IgE (FcepsilonRI), the low-affinity receptor for IgE (FcepsilonRII/CD23) and the Mac-2/IgE-binding protein (epsilonBP), have previously been found on human neutrophils. We have previously shown that specific allergens are able to activate functional responses by neutrophils from allergic patients sensitized to those allergens. Neutrophils are present in the sites of allergic inflammation. The primary (azurophilic) granules of neutrophils contain a variety of enzymes that might potentiate inflammation, such as myeloperoxidase (MPO). It is not known whether specific allergens are able to elicit MPO release by neutrophils from allergic patients. METHODS: Neutrophils were challenged in vitro with the specific allergen that produced clinical symptoms in asthmatic patients. Also, the cells were challenged with allergens that the patients were not sensitive to. Neutrophils from normal subjects were also challenged with allergens. RESULTS: The in vitro challenge of neutrophils with allergens that the patients were sensitive to elicited a release of MPO by these cells. The in vitro activation of neutrophils was highly allergen-specific, in such a way that allergens other than those accounting for clinical symptoms did not evoke MPO release, and allergens were ineffective on neutrophils from healthy donors. CONCLUSION: An IgE-dependent mechanism might promote MPO release by neutrophils at allergic sites. PMID- 11422154 TI - Decreased T lymphocyte infiltration in bronchial biopsies of subjects with severe chronic obstructive pulmonary disease. AB - BACKGROUND: Studies on the inflammatory process in the large airways of patients with mild/moderate COPD have shown a prevalent T lymphocyte and macrophage infiltration of the bronchial mucosa. However, bronchial inflammation in more severe disease has not been extensively studied. OBJECTIVE: The aim of the present study was to characterize the lymphocyte infiltration in the bronchial mucosa of subjects with severe, compared to mild, COPD, and to examine the relationship between airflow limitation and T lymphocyte numbers in the bronchial mucosa. METHODS: We examined bronchial biopsies obtained from nine smokers with severe airflow limitation, nine smokers with mild/moderate airflow limitation and 14 smokers with normal lung function. Immunohistochemical methods on cryostat sections were used to assess the number of CD3+, CD4+, CD8+ cells and the number of CD3+ cells coexpressing the chemokine receptor CCR5 (CCR5+CD3+) in the subepithelium. RESULTS: Subjects with severe COPD had lower numbers of CD3+, CD8+ and CCR5+CD3+ cells than mild/moderate COPD (P < 0.012, P < 0.02 and P < 0.02, respectively) and control smokers (P < 0.015, P < 0.005 and P < 0.015, respectively). In subjects with airflow limitation the number of CD3+ and CD8+ cells was inversely correlated with the degree of airway obstruction (r = 0.59, P < 0.015 and r = 0.52, P < 0.032, respectively). CONCLUSIONS: Bronchial inflammation in severe COPD is characterized by lower numbers of CD3+ and CD8+ cells and decreased numbers of CD3+ cells coexpressing the chemokine receptor CCR5. T lymphocyte infiltration is inversely correlated with the degree of airflow limitation. PMID- 11422155 TI - Validation of the Chinese translated version of ISAAC core questions for atopic eczema. AB - BACKGROUND: The International Study of Asthma and Allergies in Childhood (ISAAC) was designed to allow international comparison of epidemiological data on atopic conditions in childhood. In so doing, further aetiological information would be obtained that in turn would provide a framework for future studies. The global ISAAC results on the prevalence of atopic dermatitis indicated a 60-fold variation recorded in different countries. Such a degree of difference may be partially due to the translated questionnaires that were not validated in all of the involved countries. OBJECTIVE: To validate the Chinese version of the ISAAC core questions for atopic eczema. METHODS: One thousand nine hundred and twenty children aged between 3 and 5 were randomly recruited from 13 kindergartens in Hong Kong. Using a dermatologist's clinical examination as the gold standard, we validated the Chinese version of the ISAAC core questions for atopic eczema. The Youden's Indexes obtained in our study were compared with those obtained in the United Kingdom's validation study. RESULTS: The Youden's Indexes obtained in our study were significantly lower than those from the United Kingdom. The low scores were likely to be due to a reduction in the sensitivity of the Chinese questionnaire, which ranged from 23.5% to 70.6%. CONCLUSION: Our findings indicate that the translated questionnaire is less effective than the English version in assessing the prevalence of atopic eczema. The indication of a low prevalence of atopic eczema among the Chinese population reported in previous studies was at least partially due to problems with the translated questionnaire. PMID- 11422156 TI - Does tobacco smoke prevent atopic disorders? A study of two generations of Swedish residents. AB - BACKGROUND: Earlier studies have given conflicting results regarding the effect of exposure to tobacco smoke on atopic sensibilization. METHODS: A cross sectional study of present and former smoking habits in relation to atopic disorders from data on 6909 young and middle-aged adults (16-49 years) and their 4472 children (3-15 years) from the Swedish Survey of Living Conditions in 1996 97. RESULTS: The prevalence of allergic asthma and allergic rhino-conjunctivitis decreased, in a dose-response manner (P = 0.03 and P = 0.004, respectively), with increasing exposure to tobacco smoke in the adult study population. This pattern was little changed when potential confounders (sex, age, education, domicile, country of birth) were entered into a multivariate analysis: the adjusted odds ratio (OR) for allergic rhino-conjunctivitis was 0.5 (0.4-0.7) for those who smoked at least 20 cigarettes a day and OR 0.7 (0.6-0.9) for those smoking 10-19 cigarettes, compared with those who reported that they never had smoked Former smokers had a tendency for a slightly lower risk: OR 0.9 (0.8-1.0). In a multivariate analysis, children of mothers who smoked at least 15 cigarettes a day tended to have lower odds for suffering from allergic rhino-conjunctivitis, allergic asthma, atopic eczema and food allergy, compared to children of mothers who had never smoked (ORs 0.6-0.7). Children of fathers who had smoked at least 15 cigarettes a day had a similar tendency (ORs 0.7-0.9). CONCLUSIONS: This study demonstrates an association between current exposure to tobacco smoke and a low risk for atopic disorders in smokers themselves and a similar tendency in their children. There is a need for further studies with a prospective design to certify the causal direction of this association. Smoking habits and atopic disorder in parents should not be considered independent variables in epidemiological studies of the connection between exposure to tobacco smoke and atopy in children. PMID- 11422157 TI - In vitro and in vivo responses to the recombinant bovine dander allergen Bos d 2 and its fragments. AB - BACKGROUND: About one in every four cases of occupational rhinitis recorded in Finland is animal-induced. Bovine allergens are the most important in this respect and the largest patient group consists of dairy farmers. Allergen immunotherapy, if proven effective, safe and feasible, would be ideal for their treatment. The development of recombinant allergens has offered new potential therapeutic prospects. Fragments of recombinant Bos domesticus (Bos d 2) allergen could be suitable for this purpose because they are recognized by T cells but their IgE-binding capacity is attenuated. OBJECTIVE: The aim of this study was to verify whether the potential of the two fragments of recombinant Bos d 2 (corresponding to amino acids 1-131 and 81-172) to induce immediate allergic reaction in a shock organ (nose) was decreased compared to the complete recombinant allergen, which would be an advantageous property for a preparation intended for allergen immunotherapy. METHODS: The study group consisted of 10 dairy farmers with cow-induced allergic rhinitis. We used the IgE titres against native Bos d 2 measured by indirect IgE ELISA to characterize the level of sensitization and compared the IgE titres in the rhinitis patients with 12 cow sensitized asthmatic farmers and 12 healthy students. In vitro reactivity against recombinant Bos d 2 and its two fragments was studied by indirect IgE ELISA and in vivo reactivity by nasal provocation tests. RESULTS: The IgE titres against native Bos d 2 of patients with rhinitis tended to be lower than the titres of asthmatics. The healthy students did not exhibit any detectable IgE reactivity to native Bos d 2. In the patients with rhinitis, there was no statistically significant difference between IgE responses against native and recombinant Bos d 2, whereas with both in vitro and in vivo, the reactivity to both fragments of recombinant Bos d 2 was lower than the reactivity to the complete recombinant allergen. CONCLUSIONS: Due to the decreased in vivo capacity to induce immediate allergic reactions, the fragments may be better tolerated in allergen immunotherapy than the complete allergen. PMID- 11422158 TI - Fusarium solani major allergen peptide IV-1 binds IgE but does not release histamine. AB - BACKGROUND: Fusarium solani (FS) is an important allergen source afflicting 4% of the nasobronchial allergy patients. Fus s I3596*, a 65 kDa major glycoprotein allergen of FS reacts with 95% fungus sensitive patients. OBJECTIVES: To purify and characterize a potent peptide from Fus s I3596* which may be useful for therapeutic purposes. METHODS: The 65 kDa protein was sequentially cleaved with trypsin and cyanogen bromide (CNBr). The cleaved products were purified on reverse phase high performance liquid chromatography (rpHPLC) column and functionally characterized by in vitro and in vivo methods for its IgE binding and histamine release. RESULTS: The protein on cleavage showed 11 peaks (I to XI). Of these, peaks I, III, IV and V were highly allergenic as determined by IgE ELISA. These peaks were further purified and peptide IV-1 was most potent in comparison to other peptides by ELISA-inhibition. This peptide showed IgE binding but could not evoke intradermal response in Fusarium-sensitive patients. Heparinized blood challenged with peptide IV-1 does not release histamine. Preincubation of heparinized blood with peptide IV-1 and challenging with crude extract blocked histamine release in a dose dependent manner. CONCLUSION: Peptide IV-1 binds to IgE but does not release histamine, demonstrating its potential use in therapy of Fusarium-allergic patients. PMID- 11422159 TI - Measurement of personal exposure to aerosols of Nephrops norvegicus (scampi) using a monoclonal-based assay. AB - BACKGROUND: The industrial processing of scampi (Nephrops norvegicus) generates bioaerosols which can cause occupational respiratory disease in exposed individuals. The objective of this study was to develop a monoclonal antibody based immunoassay to measure scampi proteins collected using standard personal air sampling methods. METHODS: A monoclonal antibody was generated which predominantly bound to a single 97 Kd scampi protein, which we have shown to be a major allergen. This protein was shown by SDS PAGE immunoblotting to be present in various aqueous extracts from wastes produced during scampi processing. A quantitative immunoslotblot method was developed to quantify scampi from air sample filters. RESULTS: We demonstrated the utility of this method by showing a significant difference (P = 0.015) in the level of exposure of two groups of 12 workers processing scampi at a traditional open workstation (GM = 607 ng m(-3)) and at an enclosed workstation (GM = 451 ng m(-3)). CONCLUSION: We conclude that this assay can be used, in conjunction with health surveillance, to monitor exposure and the efficacy of measures to reduce exposure to airborne scampi allergen in an occupational environment. PMID- 11422160 TI - IL-5 deficiency abolishes aspects of airway remodelling in a murine model of lung inflammation. AB - BACKGROUND AND OBJECTIVES: Lung remodelling is a recognized feature of chronic asthma. In the present study, we have used IL-5-deficient mice to evaluate the role of this cytokine and eosinophilic inflammation in the initial stages of the structural changes occurring in the lung after antigen challenge. METHODS: Ovalbumin-sensitized wild type and IL-5-deficient mice were daily challenged for 5 consecutive days and killed 3 or 7 days after the last challenge to study the inflammatory and remodelling events, respectively. RESULTS: Wild type mice challenged with ovalbumin exhibited an accumulation of eosinophils in the bronchoalveolar lavage (BAL) fluid, associated with a production of BAL cellular fibronectin. Histological analysis also revealed an antigen-specific increase in epithelial and alveolar cell proliferation together with an increase in mucus producing epithelial cells. Eosinophilic infiltration and the associated lung remodelling were totally abrogated in IL-5-deficient mice. In wild type mice, treated intranasally with 1 microg of murine IL-5 for 5 consecutive days, no BAL eosinophilia and structural changes of the lungs could be observed. CONCLUSION: Our results demonstrate that eosinophil accumulation, but not IL-5 alone, plays a central role in the initial stages of the lung remodelling process and suggests that therapies directed at inhibiting eosinophilic inflammation may be beneficial in treating chronic asthma. PMID- 11422161 TI - A3 receptors mediate rapid inflammatory cell influx into the lungs of sensitized guinea-pigs. AB - BACKGROUND: Inhaled adenosine causes bronchoconstriction in asthmatics and may modulate inflammatory cell activity. Elevated adenosine levels occur in the lungs after antigen challenge of asthmatics. OBJECTIVES: The aim of this study was to investigate whether the bronchoconstrictor effects of the adenosine derivative, 5'-AMP, were associated with altered migration of inflammatory cells into the airways using a sensitized atopic guinea-pig model previously shown to display a bronchoconstrictor response. Comparisons were made with the effects of inhaled antigen. METHODS: Airway responses of conscious sensitized guinea-pigs to inhalation exposures of 5'-AMP were determined by whole body plethysmography as the change in specific airway conductance (sGaw). Influx of leucocytes into the airways was determined by bronchoalveolar lavage (BAL). RESULTS: 5'-AMP caused bronchoconstrictor airway responses in sensitized animals. Dose-dependent infiltration of inflammatory cells into the lungs occurred 1 h after 5'-AMP exposure. No bronchoconstriction or cell influx was seen in unsensitized guinea pigs. Exposure to ovalbumin (OA) also caused influx of inflammatory cells. Twenty four hours after an OA exposure, 5'-AMP produced no bronchoconstriction. The P1 receptor antagonists, 8-PT and 8-SPT, inhibited the 5'-AMP-induced bronchoconstriction, indicating that the bronchoconstriction seen in sensitized animals is mediated by A1 or A2 receptors. They had no effect on the cell influx, whereas the A3 antagonist, MRS-1220, significantly inhibited cellular infiltration, suggesting mediation through A3 receptors. At 24 h after an OA challenge and accompanying the cellular influx, there was airway hyper responsiveness to the bronchoconstriction by histamine. In contrast, no hyper responsiveness to histamine was seen 1 h after 3 mM or 24 h after 300 mM 5'-AMP. CONCLUSIONS: 5'-AMP caused a rapid migration of eosinophils and macrophages into the airways only in sensitized guinea-pigs, and this was blocked by the A3 antagonist MRS-1220. This was not associated with bronchial hyper-reactivity to histamine. PMID- 11422162 TI - Novel B and T cell epitopes of chicken ovomucoid (Gal d 1) induce T cell secretion of IL-6, IL-13, and IFN-gamma. AB - BACKGROUND: Chicken ovomucoid (OM, Gal d 1) has an important role in the pathogenesis of IgE-mediated allergic reactions to hen's egg white. OBJECTIVES: The purpose of this study was to clarify the mechanisms of T cell recognition of ovomucoid using intact OM and chemically modified, characterized and homogeneous solid phase synthetic peptides covering the whole molecule. METHODS: Eighteen overlapping peptides were prepared by solid phase F-moc polyamide peptide synthesis (SPPS), characterized and high-pressure liquid chromatography (HPLC) purified. The peptides, together with intact, denatured and oxidized OM, were used to stimulate patient-derived cell cultures for mapping T cell epitopes. Proliferation responses, T cell phenotype and cytokine secretion using peripheral blood mononuclear cells (PBMC) from eight individuals and T cell lines (TCL) derived from six hen's egg-allergic patients, were examined. In addition, intact, denatured, oxidized and deglycosylated OM, as well as the peptides solely or with their keyhole limpet haemocyanin (KLH) complexes, were tested. For locating IgE and IgG B cell epitopes, seven egg-allergic patient sera and three OM-polyclonal sera were used. Healthy non-allergic individuals were included as controls. RESULTS: Seven peptides were recognized by specific IgE, while OM-specific TCL recognized 10 peptides. Six of the OM peptides were commonly recognized both by patient S-IgE and blood-derived TCL. Among those, one novel epitope, peptide OM 61-74, had the ability to bind IgE. Another peptide, OM 101-114, was recognized by IgE and IgG sera, but not by any of the TCLs. In contrast, the peptides OM 41 56, OM 71-84, OM 131-144 and OM 171-186 were exclusively T cell epitopes with no affinity to specific antibodies. Abundant TCL secretion of IFN-gamma, IL-6, IL-4, IL-13, IL-10 and TNF-alpha in response to OM stimulation indicates the contribution of Th2 as well as Th1/Th0 CD4+ cell subsets. For allergic patients moderate amounts of IFN-gamma, IL-13, and high amounts of IL-6, were secreted in response to TCL stimulation by OM peptides. High amounts of IL-6 were secreted in response to all molecular forms of OM (intact-, modified-OM and the peptides 71 84 and 51-64) when TCLs from two non-allergic donors were used. CONCLUSIONS: One novel B cell epitope (OM 61-74) and 10 T cell epitopes have been identified. The most reactive epitopes of the OM molecule comprise the motifs 1-14 to 71-84, the overlapping peptide-pairs OM 121-134 and OM 131-144 and peptides OM 161-174 and 171-186. Peptides OM 1-14 and 171-186 are the only ones capable of inducing IL-4 secretion. Only one peptide (OM 11-24) induces IL-10 secretion. Those peptides recognized as both T and B cell epitopes or only T cell epitopes, have the potential to induce T cell secretion of moderate to high amounts of IL-13, IFN gamma and particularly IL-6. PMID- 11422163 TI - The management of porphyria cutanea tarda. AB - Porphyria cutanea tarda (PCT), the commonest of all porphyrias, is usually characterized by blisters and fragility of skin in light-exposed areas. It can be clinically indistinguishable from other disorders including variegate porphyria and the diagnosis can only be made by rigorous biochemical analysis. PCT does not cause acute attacks of porphyria. It is usually an acquired condition caused by inhibition of the uroporphyrinogen decarboxylase enzyme in the liver. Hereditary haemochromatosis, hepatitis C virus infection, alcohol, oestrogens and a family history of PCT are the major risk factors for the condition and should be searched for specifically in all patients. Liver disease, including hepatocellular carcinoma, is common in patients with PCT, and should be investigated for at presentation by means of a liver biopsy where possible. Patients with severe hepatic pathology or longstanding untreated PCT need to be monitored for the development of hepatocellular carcinoma in the long term. Low dose twice weekly chloroquine is the mainstay of treatment, but venesection should be used in patients with severe iron overload or hepatitis C-related liver disease. Subsequently, long-term follow-up is needed in all patients to monitor for relapse. PMID- 11422164 TI - Improvement in psoriasis after intradermal administration of delipidated, deglycolipidated Mycobacterium vaccae (PVAC): results of an open-label trial. AB - The aim of new treatments for psoriasis is to induce extended remissions with fewer side-effects. Previous studies suggest that Mycobacterium vaccae, a harmless organism prepared as a heat-killed suspension, may induce periods of remission in some psoriasis patients after intradermal administration. To assess a more potent derivative of M. vaccae, we conducted an open-label study in which 20 patients with moderate to severe psoriasis (Psoriasis Area and Severity Index of 12-35) received two intradermal inoculations of heat-killed, delipidated, deglycolipidated M. vaccae (DD-MVAC or 'PVAC') in lesion-free deltoid skin, separated by a period of 3 weeks. Twelve weeks after the injections, 13 out of 20 patients (65%) showed marked improvement in the PASI score (> 50% reduction), three were unchanged (< 25% reduction), three had worsened (> 5% increase), and one was withdrawn from the trial because of an exfoliative flare. At 24 weeks, 13 out of 19 patients continued to show > 50% improvement that, in some, lasted for 6 months or longer. Patients classified as good responders at 12 or 24 weeks were then offered additional PVAC injections after 24 weeks if the PASI reached 8 or higher. Intra-dermal administration of PVAC was safe, well tolerated, and induced clinically significant improvement in many psoriasis patients. A randomized, double-blind, controlled study is warranted. PMID- 11422165 TI - Epstein-Barr virus-associated peripheral T-cell lymphoma in adults with hydroa vacciniforme-like lesions. AB - We describe two Korean adult patients who had necrotizing papulovesicles mainly on their faces. Skin biopsy specimens showed perivascular and periadnexal infiltrate of atypical lymphoid cells with vasculitis in the dermis and subcutaneous tissue. In situ hybridization demonstrated a latent infection of Epstein-Barr virus in the majority of lymphoid cells in the dermis. These patients were diagnosed as having T-cell lymphoma. Interestingly, large granular lymphocytosis was found in the peripheral blood of Case 2. PMID- 11422166 TI - Severe cutaneous reaction to CS gas. AB - Tear gas is used throughout the World for control of riots and civil disobedience. CS gas as used by the UK police force is issued as a 'spray' and is 5% CS in methylisobutylketone (MIBK), a potent irritant. Assaults on police officers in forces issued with CS spray have fallen significantly over the past 3 years, whilst having risen in areas without it. Thus, CS gas appears to be an effective deterrent. However, significant cutaneous reactions can occur as a result of exposure. We report a severe contact dermatitis to CS gas to highlight the clinical features. The nature of CS gas and potential cutaneous adverse reactions are discussed. PMID- 11422167 TI - Neonatal eosinophilic pustular folliculitis. AB - Eosinophilic pustular folliculitis (EPF) of infancy is a rare disorder which may begin in the neonatal period and cause considerable parental anxiety. It must be distinguished from other causes of a pustular eruption in neonates, including infection and erythema toxicum neonatorum, and rare disorders such as transient neonatal pustular melanosis, infantile acropustulosis and Langerhans' cell histiocytosis. Skin smears and occasionally skin biopsy may be necessary to reach a diagnosis. We report a case of a Caucasian child with an unusually early onset of EPF in the first day of life. We wish to emphasize the importance of recognizing this self-limiting condition in order to prevent inappropriate antimicrobial treatment. PMID- 11422168 TI - Widespread biphasic amyloidosis: response to acitretin. AB - We report a patient who presented with an intensely pruritic rash, with hyperpigmentation of the whole back and multiple hyperpigmented and hyperkeratotic papules over his forearms and lower extremities. Histologically, he showed deposits of amyloid that reacted positively with monoclonal antibodies against cytokeratins. The lesions were resistant to antihistamines and corticosteroids. Treatment with oral acitretin was started and great improvement of the lesion was noted; the patient became asymptomatic. We review the literature on the effect of retinoids in cutaneous amyloidosis. PMID- 11422169 TI - Rifampicin-induced lupus erythematosus. AB - Rifampicin has been prescribed throughout the world for over 20 years, yet only four cases of rifampicin-induced lupus erythematosus (LE) have been reported. Rifampicin-induced LE is associated with combination therapy with clarithromycin or ciprofloxacin. These drugs are all metabolized through the cytochrome P450 liver enzyme system and combined usage may lead to higher rifampicin blood levels. Drug-induced LE differs from systemic LE; cutaneous manifestations, although uncommon, are an important clue to the diagnosis. We report a case of rifampicin-induced LE presenting with florid cutaneous features. PMID- 11422170 TI - Acquired crateriform hyperkeratotic papules of the lower limbs: an unusual variant of acrokeratoelastoidosis of Costa. AB - We report a man with a 3-year-history of a striking eruption of small, 2-3 mm diameter keratotic, crateriform papules distributed symmetrically over the lower limbs. Morphologically the individual papules closely resembled those described in acrokeratoelastoidosis (AK) of Costa and in focal acral hyperkeratosis of Dowd (FAH). The case is unusual, however, in that there was dramatic involvement of the legs but the hands and feet were largely spared. Current terms used for these entities are unsatisfactory. PMID- 11422171 TI - Endemic (African) Kaposi's sarcoma presenting as a plantar tumour. AB - We present a case of the aggressive variant of African endemic Kaposi's Sarcoma (AKS) which presented as a large fungating and ulcerated plantar mass. Our patient responded extremely well to chemotherapy with no recurrence for 9 months after treatment completion. AKS is one of the most common cutaneous neoplasms in black Africans and although rare in Europe, it may be seen more frequently in the future because of the ease of international travel. The existing classification of KS into five different types (classic, African-endemic, iatrogenically immunocompromised patients, epidemic HIV-related and Mediterranean-endemic) cannot address fully the many anomalies described in the disease. The detection of a new herpes simplex-like viral DNA sequence (HHV-8) in different types of KS helps to explain some of the enigma described in this disease. PMID- 11422172 TI - Lipoatrophy semicircularis induced by trauma. AB - Lipoatrophy semicircularis is thought to be a rare condition characterized by band-like horizontal depressions of the skin typically involving the lower limbs. Previous literature has suggested that repetitive trauma to the lower limbs could explain this condition; however, no direct causal link has ever been clearly established. There have also been several reports where no preceding history of trauma could be found. In our series we report seven cases of lipoatrophy semicircularis representing over one-third of the staff in a particular office. We propose that the most logical explanation for the indentations present in these individuals is repetitive trauma to their thighs by the sharp edge of the desks. The consistent nature of the distance between the floor and the horizontal indentations on the lower limbs despite differences in height weight and body mass index support this theory. It is likely that this condition is more common than initially thought. PMID- 11422173 TI - Hand rash in trichinosis. AB - Trichinosis patients may develop peri-orbital oedema, conjunctival haemorrhages, splinter haemorrhages of the fingernails, and nonspecific skin rashes. Here, we describe an unusual hand rash noted in several patients enrolled in a treatment study for trichinosis. PMID- 11422174 TI - Psoriasis, psoriatic arthropathy and relapsing orbital myositis. AB - Orbital myositis is an inflammatory disorder of the orbital muscles causing orbital pain and restriction of eye movements. Although rare in children, it is most frequently seen after orbital trauma or as a post-infectious process. We describe a child with chronic relapsing psoriasis, juvenile psoriatic arthritis and relapsing bilateral orbital myositis. PMID- 11422175 TI - Terbinafine treatment of Trichophyton equinum infection in a child. AB - A case of tinea faciae caused by Trichophyton equinum affecting a 5-year-old boy is described. The boy had ridden a pony a month earlier and responded to a 6-week course of treatment with oral and topical terbinafine. PMID- 11422176 TI - Searching for candidate genes in the new millennium. AB - Completion of the entire sequence of the human genome is having a profound effect on the strategies biological scientists use to identify disease-associated genes. Laborious positional cloning approaches and traditional functional studies are gradually being transformed by emerging genomic and proteomic databases. Some of the exciting challenges investigators now face are the identification of new genes, determining the function of these genes, defining disease associations, and elucidating correlation between genotype and phenotype. To demonstrate how investigative methods for single-gene disorders are changing, we illustrate one possible approach in the search for the gene underlying the autosomal recessive genodermatosis, acrodermatitis enteropathica. PMID- 11422177 TI - Oxidative stress in physical urticarias. AB - The pathogenesis of the physical urticarias has not been completely defined. Indeed, different stimuli can induce similar clinical manifestations, some of which are capable of generating reactive oxygen species. In order to evaluate whether the generation of an oxidative stress response could be a common pathogenetic mechanism of the disease, we have determined the profile of a number of chemical and enzymatic antioxidants in blood samples from a group of patients with physical urticarias. Compared with controls, a systemic imbalance of the antioxidants was detected in the patient group with a decrease of both plasma vitamin E and cellular catalase and glutathione peroxidase activities along with an increase of superoxide dismutase activity. Moreover, an increase in the percentage of plasma polyunsaturated fatty acids, as a target for peroxidative damage, was also observed. These alterations may lead to an increased percentage of peroxidable compounds in skin and to the intracellular generation of reactive oxygen species and could therefore provide one possible explanation for the patients' urticarial response to stimuli. Even if the alteration of the antioxidant status is secondary to changes in cytokine or complement activation, our results suggest a common biochemical profile in patients with different forms of physical urticaria. PMID- 11422178 TI - Phosphatidylinositol-specific-phospholipase C cleaves urokinase plasminogen activator receptor from the cell surface and leads to inhibition of pemphigus-IgG induced acantholysis in DJM-1 cells, a squamous cell carcinoma line. AB - We showed previously that pemphigus IgG enhanced both the activity of urokinase plasminogen activator (uPA) in cultured cells and the expression of its receptor (uPAR) on uPA-binding keratinocytes. In the present study, to clarify whether uPAR and uPA-activated plasmin are actually involved in the blistering process after pemphigus IgG binding to the cell surface, we examined the effects of the following on uPAR expression and on cell-cell detachment in DJM-1 cells, a squamous cell carcinoma line: (i) phosphatidylinositol-specific phospholipase C (PI-PLC) - which releases uPAR from the membrane surface into the culture medium by cleaving the glycosylphosphatidylinositol anchor thus inhibiting uPAR activity, and (ii) uPA inhibitors (tranexamic acid, aprotinin, p-aminobenzonic acid and dexamethasone). Preincubation with PI-PLC decreased dramatically the pemphigus IgG-induced uPAR expression in a dose-dependent manner, and inhibited pemphigus IgG-induced cell-cell detachment at 10 microg/mL. On the other hand, tranexamic acid (15 mM) inhibited pemphigus IgG-induced cell-cell detachment without reduction of uPAR expression, although aprotinin, p-aminobenzonic acid and dexamethasone failed to alter either of these parameters. Although uPAR expression on the pemphigus IgG-bound cell surface and uPA activation may contribute significantly to the pathogenesis of acantholysis in pemphigus, the mechanisms are complicated and should be defined further. PMID- 11422179 TI - Keratinocyte adhesion and the missing link: from Dowling-Meara to Hay-Wells. St John's Hospital Dermatological Society Annual Oration 2000. AB - Maintaining a protective barrier against the environment is an essential function of normal skin. Critical to this role are several structural proteins and glycoproteins that contribute to adhesive junctions linking adjacent keratinocytes and basal keratinocytes to the underlying dermis, as well as other regulatory proteins involved in aspects of epidermal development, differentiation and proliferation. Inherited abnormalities in the genes that encode these components may give rise to a range of genodermatoses, many of which are characterized structurally by a 'missing' or perturbed adhesive link and clinically by congenital skin blistering. This oration reviews some of the original clinical descriptions and observations made in this field, as well as providing an update on the corresponding recent molecular discoveries. The emphasis is on contributions made by past and present members of the St John's Hospital Dermatological Society. PMID- 11422182 TI - Psoriasis--epidemiology and clinical spectrum. AB - Despite psoriasis being a common skin disease, there are still a number of unanswered questions. One of these is the prevalence of the disease, as there is a lack of specific data, with the majority of studies reporting estimates only. Population based studies are rare and longitudinal observations on changing prevalence rates are lacking. This contrasts with other T-cell mediated autoimmune diseases where the number of those affected is rising. Epidemiological studies revealed that a distinct group of diseases is quite frequently associated with psoriasis, e.g. arthritis, colitis, diabetes and hypertension. In contrast, atopic dermatitis and allergies are less frequently seen compared to normal rates of occurrence. As the psoriatic immune response pattern relates to activated Th-1 cells, psoriasis and atopic dermatitis appear to be mutually exclusive due to the Th-1/Th-2 dichotomy. PMID- 11422183 TI - Genetic aspects of psoriasis. AB - Epidemiological studies indicate that genes play an important role in the pathogenesis of psoriasis. Multiple genes are likely to be involved, interacting not only with each other but also with the environment to cause disease expression. Molecular genetic studies indicate that there are multiple susceptibility loci present throughout the human genome. It is clear that a gene or genes of major impact on psoriasis is present on chromosome 6 within the major histocompatibility complex (MHC). Linkage disequilibrium studies indicate this gene to reside within a 300 kb interval centred around the centromeric end of class I MHC. Known candidate genes in this region are HLA-C, corneodesmosin and HCR, although novel genes, as yet unknown, may also exist. There is accumulating evidence that HLA-C is not itself the causative gene but rather a marker for it. Identification of the genes involved in psoriasis susceptibility will represent a step forward in our understanding of the disease and our future ability to help patients with psoriasis. PMID- 11422184 TI - Psoriasis vulgaris--a sterile antibacterial skin reaction mediated by cross reactive T cells? An immunological view of the pathophysiology of psoriasis. AB - The understanding of the pathogenesis of psoriasis vulgaris has advanced significantly since the therapeutic efficacy of immunosuppressive drugs has drawn attention to the role of immune mechanisms in psoriasis manifestation. Today, the results of many experimental studies provide evidence that psoriasis is largely a T-cell mediated disorder. It may result from antigen-specific activation of T cells in the skin of genetically predisposed individuals. These T cells apparently have a particular functional differentiation and promote the psoriatic skin changes by secreting a certain set of cytokines. Based on the fact that streptococcal throat infections are a trigger of guttate psoriasis, the putative psoriatic antigens are assumed to be in keratinocyte proteins that share structural homologies with streptococcal proteins and thus induce cross-reactive responses of antibacterial T cells against skin components. Together with the particular phenotype of psoriatic skin lesions these findings suggest that psoriasis represents a sterile antibacterial tissue reaction, which is mediated by streptococci-specific T cells that cross-react against epidermal autoantigens. PMID- 11422185 TI - Pathogenesis of psoriatic arthritis. AB - Psoriasis and psoriatic arthritis have been recognized for more than 100 years. Neither the link between psoriasis of the skin or nails and arthritis of the joints nor the pathogenesis of either condition alone or in combination has yet been explained. Our understanding of the mechanisms of inflammation of the skin and the joints has improved over the past 30 years and there are some interesting common threads of knowledge that may bring us closer to understanding psoriasis and psoriatic arthritis. This article will explore further the areas of immunogenetics, infection, autoimmunity, vascular morphology/angiogenesis, trauma and the nervous system in respect of psoriasis and psoriatic arthritis highlighting in particular those aspects that previously have not received due reference. PMID- 11422186 TI - Psychological influences in psoriasis. AB - It has long been recognized that living with a chronic condition, such as psoriasis, can have considerable impact on the individual concerned. In turn there is increased understanding that the psychological distress encountered as a result of this experience can have implications for the course of the disease. This short review takes this as a starting point and reviews psychological influences in psoriasis. The nature of the link between psoriasis and stress is explored and its implications for the patient are discussed in psychological and clinical terms. There seems little doubt that stress, either environmental or psoriasis induced, impacts on our patients and has important implications for the management of psoriasis. PMID- 11422187 TI - Phototherapy for psoriasis. AB - This review covers the current practice of phototherapy with ultraviolet (UV) radiation without sensitizers and of psoralen photochemotherapy (PUVA) in the treatment of psoriasis. Both treatment modalities are well established in today's therapeutic armamentarium. Phototherapeutic regimens use repeated controlled UV exposures to alter cutaneous biology, aiming to induce remission of skin disease. Although UVB has been used for a longer time than PUVA, the latter has been evaluated and validated in a more detailed and coordinated fashion. PMID- 11422188 TI - Dermatology day care treatment centres. AB - Day care treatment centres provide the best solution for the treatment of most patients with psoriasis. The centre is not only ideal for treatment but has other roles, such as education of patients and nurses. The specialist dermatology nurse is the key to success. Out patient treatment of psoriasis is less expensive than in patient treatment. The development of a treatment centre should be seen as an additional facility and not as a substitute for in patient beds. PMID- 11422189 TI - Therapeutic strategies: rotational therapy and combinations. AB - Long-term management of psoriasis requires an individualized approach. Some treatments such as calcipotriol, methotrexate and acitretin may be used as maintenance treatment for many months. However, most anti-psoriasis treatments should be prescribed for restricted periods of time. Rotational treatment is a practical approach to reduce the cumulative toxicity of anti-psoriasis treatments. The selection of a treatment is based on the clinical presentation of psoriasis and whether contraindications might exist. Combination treatment is another approach, which is used by the majority of patients. Useful combinations are calcipotriol-acitretin, calcipotriol-cyclosporin, calcipotriol-PUVA, calcipotriol-topical corticosteroids, dithranol-UVB, dithranol-tar, coaltar-UVB, acitretin-UVB and acitretin-PUVA. Combinations which are contraindicated are coaltar-PUVA, UVB-cyclosporin, PUVA-cyclosporin and methotrexate-acitretin. Combined use of UVB-methotrexate, UVB-PUVA; PUVA-methotrexate; methotrexate cyclosporin and cyclosporin-acitretin require careful monitoring and might be helpful in patients with severe and recalcitrant psoriasis. Depending on the individual presentation of psoriasis, previous anti-psoriatic treatments and side effects, treatment adjustments are made. PMID- 11422190 TI - Advances in systemic therapy for psoriasis. AB - Severe cases of psoriasis and psoriasis arthritis require systemic treatment. Although a number of established drugs are in clinical use, there is a need for new compounds with an improved risk-benefit ratio with a major emphasis on long term safety. Furthermore, patients with moderate psoriasis ask for systemic drugs to make therapy easier and to avoid excessive local treatments. This article aims to give a brief overview about new drugs or groups of compounds together with an evaluation of their present status in the treatment of psoriasis and their future role with particular respect to efficacy, tolerability, safety and usability. PMID- 11422191 TI - From black magic to science: understanding the rationale for the use of intravenous immunoglobulin to treat inflammatory myopathies. PMID- 11422192 TI - Slow recovery of follicular B cells and marginal zone B cells after chemotherapy: implications for humoral immunity. AB - Although most chemotherapeutic agents are known to cause primarily reduction or suppression of immune responses, surprisingly little is known about the influence of cytostatic agents on lymphoid tissue compartments such as the splenic marginal zone. The marginal zone plays an important role in the defence against encapsulated bacteria, which are potential candidates for postchemotherapeutic infections. We studied the effect of three different cytostatic agents (cisplatin, methotrexate, and cyclophosphamide) on B cell subpopulations in a rat model. Rats received a single dose of a single cytostatic agent and were sacrificed at different time points after treatment. Bone marrow, blood, mesenteric lymph nodes and spleens were analysed by flow-cytometry and immunohistochemistry. All three cytostatic agents showed severe bone marrow depression. CP and MTX showed only mild reduction of cell populations in the spleen. CyPh showed a severe reduction of recirculating follicular B (RF-B) cells and marginal zone B (MZ-B) cells. At day 24 most populations were already recovered, but RF-B cells and MZ-B cells were still reduced. The reduction of the marginal zone and late recovery may imply that, beside the overall increased infection risk due to neutropenia, patients treated with chemotherapy are at risk for developing infections from encapsulated bacteria for a considerable period of time after treatment, extending beyond the period of bone marrow depression. PMID- 11422193 TI - Membrane cofactor protein (MCP; CD46) expression in transgenic mice. AB - Human membrane cofactor protein (MCP; CD46) is a widely distributed complement regulator. In the mouse, expression of MCP is largely restricted to the testis while a related, widely expressed protein (Crry) appears to perform MCP's (CD46) regulatory activity. We have developed two mouse strains transgenic for human MCP (CD46) utilizing an approximately 400 kb YAC clone carrying the complete gene. A third mouse strain was generated using an overlapping YAC clone isolated from a second library. The expression of human MCP (CD46) in these mouse strains was characterized by immunohistochemistry, FACS, Western blotting and RT-PCR. No differences were detected in the isoform pattern or distribution among the three strains, although the expression level varied according to how many copies of the gene were integrated. The expression profile closely mimicked that observed in humans, including the same pattern of isoform expression as the donor. In addition, tissue-specific isoform expression in the kidney, salivary gland and brain paralleled that observed in man. The transgenic mice expressed low levels of MCP (CD46) on their E, in contrast to humans but in line with most other primates. These mice should be a useful tool to analyse tissue-specific expression, to establish animal models of infections and to characterize the role of MCP (CD46) in reproduction. PMID- 11422194 TI - Pregnancy induces nonimmunoglobulin insulin-binding activity in both maternal and cord blood serum. AB - To evaluate whether pregnancy has any effect on insulin antibody levels and to test the concordance between a conventional radioimmunoassay and a new microassay for the detection of insulin antibodies, insulin antibodies were analysed in 104 mothers in early pregnancy and at delivery and in their newborn infants. Thirty eight of the mothers had type 1 diabetes. The concordance between the assays was high in the samples taken in early pregnancy (95%), but substantially lower in the samples taken at delivery (40%) and in the cord blood samples (68%). A considerable proportion of the mothers at delivery, especially the unaffected mothers (71%), and the newborn infants of the unaffected mothers (32%) were positive for insulin antibodies in the conventional assay but not in the microassay. Insulin antibody levels increased in the mothers, significantly so in the unaffected mothers (P < 0.001), during pregnancy in the conventional assay, whereas in the microassay they decreased significantly (P < 0.01) in affected mothers and remained negative in the unaffected mothers. Since immune complexes are precipitated with protein A specific for IgG in the microassay and with polyethylene glycol lacking specificity for immunoglobulins in the conventional assay, our data indicate that insulin antibody levels decrease on average during pregnancy and that the increasing non-IgG anti-insulin activity observed in the conventional assay is induced by pregnancy and is present in both the maternal and the foetal circulation. PMID- 11422195 TI - Fas/FasL mediated apoptosis of thyrocytes in Graves' disease. AB - We examined in the present study the possible involvement of Fas and its ligand (FasL) in the process of Graves' disease. Immunohistochemical analysis showed that few normal thyrocytes expressed Fas but many thyrocytes in Graves' disease expressed this molecule. The percentage of FasL-positive thyrocytes in Graves' thyroids was, however, less than in normal thyroids. Several apoptotic thyrocytes and infiltrating mononuclear cells (MNCs) were detected scattered throughout Graves' thyroid tissues and abundant proliferating cell nuclear antigen (PCNA) positive thyrocytes were present. Apoptotic cells, as well as PCNA-positive cells, were scarcely detectable in normal thyroid glands, however. In vitro treatment of thyrocytes by IL-1beta a cytokine found to be expressed in Graves' thyroid glands, increased Fas but reduced FasL expression. IL-1beta-stimulated thyrocytes became sensitive to apoptosis by anti-Fas IgM monoclonal antibody (mAb). Activated T cells, which strongly expressed FasL, showed cytotoxic activity toward IL-1beta-stimulated thyrocytes but not toward unstimulated thyrocytes. This cytotoxic activity involved the Fas/FasL pathway. Importantly, unstimulated thyrocytes could kill activated, but not resting, T cells. IL-1beta stimulated thyrocytes, with down-regulated FasL expression, could not efficiently kill activated T cells. The cytotoxic activity of unstimulated thyrocytes toward activated T cells was inhibited by anti-FasL mAb. Interestingly, unstimulated thyrocytes induced apoptosis in IL-1beta-stimulated thyrocytes but not in unstimulated thyrocytes. These interactions were also blocked by anti-FasL mAb. Our results suggest that the apoptotic cell death of both thyrocytes and infiltrating MNCs found in Graves' thyroid glands is regulated by IL-1beta through Fas/FasL interactions. PMID- 11422196 TI - Colonic epithelial cells induce endothelial cell expression of ICAM-1 and VCAM-1 by a NF-kappaB-dependent mechanism. AB - Epithelial cells are positioned in close proximity to endothelial cells. A non contact coculture system was used to investigate whether colonic epithelial cells activated with various cytokines are able to provide signals that can modulate ICAM-1 and VCAM-1 expression on endothelial cells. Coculture of human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC-1) with TNF-alpha/IFN-gamma-stimulated human colon epithelial cell lines led to a significant up-regulation of endothelial ICAM-1 and VCAM-1 expression. Increased ICAM-1 and VCAM-1 expression by endothelial cells was accompanied by an increase in endothelial cell NF-kappaB p65 and NF-kappaB-DNA-binding activity. Inhibition of endothelial NF-kappaB activation using the proteosome inhibitors MG-132 and BAY 11-7082 resulted in a significant decrease of ICAM-1 expression, indicating an important role for NF-kappaB in this response. This cross-talk may represent a biological mechanism for the gut epithelium to control the colonic inflammatory response and the subsequent immune cell recruitment during inflammation. PMID- 11422197 TI - Mechanism of NK cell activation induced by coculture with dendritic cells derived from peripheral blood monocytes. AB - Dendritic cells (DCs) have been regarded as one of the effective antigen presenting cells, but the relationship between DCs and lymphocytes, in particular natural killer (NK) cells, remains unclear. In this study, we evaluated how DCs interact with both lymphocytes and NK cells using a coculture system. The number of lymphocytes increased significantly when cocultured with DCs (1.8-fold increase). In particular, the proliferation of NK cells was prominent. Furthermore, the coculture of DCs with lymphocytes induced a marked increase in IL-12 and IFN-gamma secretion. When contact between the DCs and lymphocytes was prevented, the secretion of both IL-12 and IFN-gamma was markedly reduced. IFN gamma production was completely blocked by an anti-IL-12 antibody, indicating that IFN-gamma secretion was dependent on IL-12 secretion. The stimulating effect of the DCs on the proliferation of the lymphocytes was partially suppressed by anti-IL-12 antibodies, and was completely attenuated when cellular contact was prevented. Furthermore, the NK cell proliferation induced by coculture with DCs was significantly blocked by the inhibition of the interaction of either CD40 CD40L or CD28-B7 molecule. The coculture with DCs enhanced NK activity by 40%, and this was partially suppressed by anti-IL-12 antibodies and was completely blocked by the inhibition of cell-to-cell contact. These results indicate that the activation of NK cells by DCs is partially mediated by IL-12 secretion, and that direct contact between DCs and NK cells play a major role in this response. PMID- 11422198 TI - Differential recognition of obligate anaerobic bacteria by human mannose-binding lectin. AB - Deficiency of the innate, humoral immune component mannose-binding lectin (MBL) predisposes individuals to a variety of infections, but the importance of MBL in infection by anaerobes has not been addressed. The attachment of MBL to a wide range of anaerobic bacteria associated with human disease and colonization was surveyed. The results suggest that for the species we examined, resistance to MBL binding may be associated with organisms that are more commonly pathogenic and that MBL binding to some bacteria may be phase variable. PMID- 11422199 TI - Differential requirement for interferon-gamma to restrict the growth of or eliminate some recently identified species of nontuberculous mycobacteria in vivo. AB - In recent years, a number of newly identified species of the genus Mycobacterium (M.) have been isolated from tissues of both immunocompetent and immunocompromised patients, e.g. M. celatum, M. intermedium, M. interjectum, M. bohemicum, M. conspicuum, M. confluentis, M. heidelbergense, M. lentiflavum, and M. branderi. Little is known about their in vivo virulence characteristics and the host factors predisposing to infection with these strains. In an effort to elucidate the pathogenesis of these nontuberculous mycobacterial species, BALB/c and syngeneic IFNgamma-deficient (GKO) mice were intravenously infected with 106 colony forming units of each of these species, and bacterial growth in infected organs and the development of splenomegaly and granulomatous liver lesions were examined for a period of 3 months. Based on their in vivo virulence, mycobacterial strains could be divided into three major groups: (i) Most species examined either grew progressively or persisted at plateau levels in the livers and spleens of immunocompetent mice, and their growth was increased in GKO mice. (ii) M. heidelbergense, M. intermedium and another species not officially accorded separate taxonomical status were eliminated in BALB/c mice, but persisted in GKO mice. (iii) M. confluentis, M. lentiflavum and another novel species were eradicated even in the absence of IFNgamma. Nontuberculous mycobacterial species differed widely in their capacity to induce splenomegaly and lymphadenopathy in GKO mice. In conclusion, IFNgamma is a crucial determinant of infection outcome with most, but not all opportunistic mycobacterial species. PMID- 11422200 TI - Effects of recombinant granulocyte-colony stimulating factor administration during Mycobacterium avium infection in mice. AB - Granulocyte colony-stimulating factor (G-CSF) administration in vivo has been shown to improve the defence mechanisms against infection by different microbes. Here we evaluated a possible protective role of this molecule in a mouse model of mycobacterial infection. The administration of recombinant G-CSF promoted an extensive blood neutrophilia but failed to improve the course of Mycobacterium avium infection in C57Bl/6 or beige mice. G-CSF administration also failed to improve the efficacy of a triple chemotherapeutic regimen (clarithromycin + ethambutol + rifabutin). G-CSF treatment did not protect interleukin-10 gene disrupted mice infected with M. avium. Spleen cells from infected mice treated with G-CSF had a decreased priming for antigen-specific production of interferon gamma compared to control infected mice. Our data do not substantiate previous reports on the protective activity of G-CSF in antimycobacterial immunity using mouse models. PMID- 11422201 TI - Expression of perforin on HIV-1-specific CD8+ lymphocytes after immunization with a gp120-depleted, whole-killed HIV-1 immunogen. AB - We examined HIV-1 antigen specific intracellular expression of perforin on CD4+ and CD8+ lymphocytes in subjects with chronic HIV-1 infection on antiviral drug therapy after immunization with a gp120-depleted, whole killed HIV-1 immunogen (inactivated, gp120-depleted HIV-1 in IFA, REMUNE). Based upon previous results, we hypothesized that the restoration of adequate T helper immune responses by vaccination against HIV-1 could result in the augmentation of CD8+ lymphocyte immune responses measured as perforin expression. In the current study we observed an increase in the frequency of perforin in CD8+ lymphocytes in HIV infected individuals immunized with a gp120-depleted HIV-1 immunogen while on antiviral drug therapy. Furthermore, the frequency of HIV-specific CD8+ perforin expressing cells correlated with the T helper immune response as measured by the lymphocyte proliferative response (LPR). The induction of such responses with immunization may have direct antiviral consequences and is being studied in ongoing clinical trials. PMID- 11422202 TI - Induction and abrogation of LACK reactive cells in the evolution of human leishmaniasis. AB - Peripheral blood mononuclear cells (PBMC) from cutaneous leishmaniasis patients with ongoing Leishmania aethiopica infection and individuals cured/under treatment from L. infantum or L. donovani infection were stimulated in vitro with LACK, the Leishmania homologue of receptors for activated C kinase. The LACK protein is conserved in related leishmanial species and is expressed both in the promastigote and amastigote stages of Leishmania. Our results show that LACK induced marked NK and some CD8+ cell proliferation in PBMC from cutaneous leishmaniasis patients with active disease. These responses were coupled with high levels of IFN-gamma and IL-10 production. At the concentration tested, the proliferative responses to freeze-thawed Leishmania antigen (Ft-Leish) were higher, while the levels of IFN-gamma were consistently lower than that of LACK. Although cells from individuals cured of leishmaniasis could respond to whole Leishmania lysate by proliferation and IFN-gamma production, there was no evident response to LACK. Ethiopian controls tested at the same time also showed LACK induced proliferation with IFN-gamma and IL-10 responses. Thus LACK reactivity in terms of proliferation and cytokine induction were present in cells from some healthy donors and most of the patients with active lesions, while this response was absent in individuals cured of L. infantum or L. donovani leishmaniasis. Since cure from leishmaniasis often results in life-long protection, and active but not cured patients showed in vitro responses to LACK stimulation, questions arose as to how this highly immunodominant molecule functions during human leishmanisasis. Some possible mechanisms are discussed. PMID- 11422203 TI - Reactive oxygen intermediates, nitrite and IFN-gamma in Indian visceral leishmaniasis. AB - Reactive oxygen intermediates (ROI), nitrite and interferon-gamma (IFN-gamma) production were investigated at different times during treatment in 10 patients with visceral leishmaniasis (VL). Hydrogen peroxide (H2O2), superoxide (O2-) and IFN-gamma production by cultured monocytes from patients with active VL were significantly lower compared with the healthy controls. In contrast, nitrite levels in the supernatants from monocyte cultures of VL patients were comparable to healthy controls and increased significantly during antileishmanial therapy. On day 20 of treatment, a significant increase in the release of H2O2, O2- and IFN-gamma was observed. However, at follow-up, 4 months after the end of treatment, the production of H2O2, O2-, IFN-gamma and nitrite had declined significantly. Thus, the impairment in hydrogen peroxide and superoxide production suggests that down-regulation of these mediators may be involved in the reduced killing of parasites by monocytes of active VL patients. Furthermore, the monocytes regained respiratory burst activity as the antileishmanial therapy progressed, suggesting that an immune-based mechanism is involved in successful drug therapy. PMID- 11422204 TI - Anti-galectin-1 autoantibodies in human Trypanosoma cruzi infection: differential expression of this beta-galactoside-binding protein in cardiac Chagas' disease. AB - The pathogenesis of Chagas' disease has been subject of active research and still remains to be ascertained. Galectin-1 (Gal-1), a member of a conserved family of animal beta-galactoside-binding proteins, localized in human heart tissue, has been suggested to play key roles in immunological and inflammatory processes. In the present study we demonstrated the occurrence of anti-Gal-1 autoAb in sera from patients in the acute and chronic stages of Chagas' disease (ACD and CCD) by means of ELISA and Western blot analysis. We found a marked increase in the level and frequency of Ig E anti-Gal-1 antibodies in sera from patients with ACD, but a low frequency of Ig M anti-Gal-1 immunoreactivity. Moreover, Ig G immunoreactivity to this beta-galactoside-binding protein was found to be correlated with the severity of cardiac damage in CCD, but was absent in nonrelated cardiomyopathies. We could not detect immunoreactivity with Trypanosoma cruzi antigens using a polyclonal antibody raised to human Gal-1 and no hemagglutinating activity could be specifically eluted from a lactosyl-agarose matrix from parasite lysates. Moreover, despite sequence homology between Gal-1 and shed acute phase antigen (SAPA) of T. cruzi, anti-Gal-1 antibodies eluted from human sera failed to cross-react with SAPA. In an attempt to explore whether Gal-1 immunoreactivity was originated from endogenous human Gal-1, we finally investigated its expression levels in cardiac tissue (the main target of Chagas' disease). This protein was found to be markedly upregulated in cardiac tissue from patients with severe CCD, compared to cardiac tissue from normal individuals. PMID- 11422205 TI - Pentoxifylline inhibits the synthesis and IFN-gamma-inducing activity of IL-18. AB - The effect of phosphodiesterase-inhibiting anti-inflammatory drug pentoxifylline (PTX) on LPS-induced IL-18 synthesis and IL-18-mediated IFN-gamma-induction were investigated. In a dose-dependent manner PTX inhibited production of IL-18 in LPS treated cultures of murine spleen cells and bone marrow-derived macrophages. Similarly, PTX treatment significantly reduced blood IL-18 levels and expression of spleen IL-18 mRNA in LPS-challenged mice. The inhibitory effect of PTX was specific for IL-18, since LPS-induced IL-12 p40 release was not suppressed either in splenocyte cultures or blood of LPS-injected animals. Synergistic induction of IFN-gamma by combined IL-12/IL-18 treatment was also inhibited by PTX in vitro and in vivo. Experiments with IL-12 pretreatment of splenocytes, followed by IL 18 stimulation, revealed that PTX suppressed both IL-12 and IL-18 signals responsible for IFN-gamma induction. These results suggest that interference with IL-18 synthesis and IFN-gamma-inducing activity might contribute to anti inflammatory actions of PTX. PMID- 11422206 TI - Intravenous immunoglobulin prevents experimental autoimmune myositis in SJL mice by reducing anti-myosin antibody and by blocking complement deposition. AB - High-dose intravenous immunoglobulin (IVIG) therapy has been effective in many autoimmune and systemic inflammatory diseases including polymyositis (PM) and dermatomyositis (DM). In the present study we evaluated the efficacy of IVIG using experimental models of PM and DM. An experimental autoimmune myositis (EAM) model was produced in SJL/J mice by an immunization with rabbit myosin B (MB) fraction. In this model, the plasma level of anti-MB antibody was elevated, and mouse IgG and complement C3 were deposited in the muscle fibres. Administration of IVIG dose-dependently reduced the incidences of necrotic and inflammatory changes in the skeletal muscle. IVIG treatment also decreased the elevation of anti-MB antibody level, as well as the deposition of IgG and C3. We next evaluated the effect of IVIG in adoptive EAM mice made by an intravenous injection of lymph node cells previously stimulated with MB. Adoptive EAM mice showed similar lesions in skeletal muscle as EAM mice and IVIG inhibited the lesion development. In vitro experiments demonstrated that IVIG inhibited complement-mediated lysis of human erythrocytes sensitized with anti-human erythrocyte antibodies. The binding of C1q, C4 and C3 to the same cells was also inhibited by IVIG. Taken together these findings suggest that IVIG prevents the development of myositis in EAM and adoptive EAM models by several mechanisms, such as reducing anti-myosin antibody and by blocking complement activation. Our present findings might account for the clinical efficacy of IVIG in PM and DM patients. PMID- 11422207 TI - Regulation of LPS induced IL-12 production by IFN-gamma and IL-4 through intracellular glutathione status in human alveolar macrophages. AB - Interleukin-12 (IL-12) is secreted from monocytes and macrophages; it exerts pleiotropic effects on T cells and natural killer (NK) cells, and stimulates interferon-gamma (IFN-gamma) secretion. Glutathione tripeptide regulates the intracellular redox status and other aspects of cell physiology. We examined whether IFN-gamma and IL-4 affect the balance between intracellular reduced glutathione (GSH) and oxidized (GSSG) glutathione, as this may affect IL-12 production in human alveolar macrophages (AM). We used both AM from healthy non smokers obtained by bronchoalveolar lavage and the monocytic THP-1 cell line in this study. Incubation of AM for 2 h with the GSH precursor N-acetylcysteine (NAC) increased the intracellular GSH/GSSG ratio, and enhanced lipopolysaccharide (LPS)-induced IL-12 secretion by AM. In THP-1 cells, NAC increased the GSH/GSSG ratio and the expression of LPS-induced IL-12 mRNA, whereas L-buthionine-[S,R] sulphoximine (BSO) decreased these. NAC and BSO offset their own effects on the intracellular GSH/GSSG ratio and the expression of LPS-induced IL-12 mRNA. Furthermore, exposure of AM to the helper T cell type 1 (Th1) cytokine IFN-gamma or the helper T cell type 2 (Th2) cytokine IL-4 for 72 h increased and decreased the GSH/GSSG ratio, respectively. Lipopolysaccharide (LPS)-induced secretion of IL-12 in AM was enhanced by IFN-gamma but inhibited by IL-4. These results suggest that IFN-gamma and IL-4 oppositely affect the GSH/GSSG balance, which may regulate IL-12 secretion from AM in response to LPS. PMID- 11422208 TI - Identification of structural variations in the carboxyl terminus of Alzheimer's disease-associated beta A4[1-42] amyloid using a monoclonal antibody. AB - The accumulation of amyloid plaques and amyloid congophilic angiopathy (ACA) in the brains of affected individuals is one of the main pathological features of Alzheimer's disease. Within these deposits, the beta A4 (Ass) polypeptide represents a major component with the C-terminal 39-43 amino acid variants being most abundant. Using a mouse IgG1 MoAb produced by hybridoma beta A4[35-43]-95.2 3B9, which reacts with the epitope is defined by the amino acid residues beta A438[GVV]40, this study has identified a unique conformation within the carboxyl terminus of human beta A4[1-42]. Although the beta A438[GVV]40 sequence is present within the C-termini of human beta A4[1-40] and beta A4[1-43] and the beta A4-containing region of human APP, the beta A4[35-43]-95.2 3B9 MoAb (designated MoAb 3B9) does not bind these polypeptides, demonstrating a high degree of specificity for the beta A438[GVV]40 epitope as presented within the beta A4[1-42] sequence. The beta A4[1-42] epitope bound by MoAb 3B9 is sensitive to heating (100 degrees C for 5 min) and is denatured by SDS but not by oxidative radio-iodination of beta A4 or by adsorption to plastic surfaces or nitrocellulose. The recognition of beta A4 plaque deposits and ACA by MoAb 3B9 within formalin-fixed sections of human AD brain demonstrates the potential of these antibodies for investigating the role of the unique beta A4[1-42] conformation in the development of Alzheimer's disease. PMID- 11422209 TI - Altered phenotype and function of blood dendritic cells in multiple sclerosis are modulated by IFN-beta and IL-10. AB - Multiple sclerosis (MS) is assumed to result from autoaggressive T cell-mediated immune responses, in which T helper type 1 (Th1) cells producing cytokines, e.g. IFN-gamma and lymphotoxin promote damage of oligodendrocyte-myelin units. Dendritic cells (DCs) as potent antigen presenting cells initiate and orchestrate immune responses. Whether phenotype and function of DCs with respect to Th1 cell promotion are altered in MS, are not known. This study revealed that blood derived DCs from MS patients expressed low levels of the costimulatory molecule CD86. In addition, production of IFN-gamma by blood mononuclear cells (MNCs) was strongly enhanced by DCs derived from MS patients. IFN-beta and IL-10 inhibited the costimulatory capacity of DCs in mixed lymphocyte reaction (MLR) and showed additive effects on suppression of IL-12 production by DCs. Correspondingly, DCs pretreated with IFN-beta and IL-10 significantly suppressed IFN-gamma production by MNCs. IFN-beta in vitro also upregulated CD80 and, in particular, CD86 expression on DCs. In vitro, anti-CD80 antibody remarkably increased, while anti CD86 antibody inhibited DC-induced IL-4 production in MLR. We conclude that DC phenotype and function are altered in MS, implying Th1-biased responses with enhanced capacity to induce Th1 cytokine production. In vitro modification of MS patients' DCs by IFN-beta and IL-10 could represent a novel way of immunomodulation and of possible usefulness for future immunotherapy of MS. PMID- 11422210 TI - Differential responses of CD45+ve T-cell subsets to MBP in multiple sclerosis. AB - The proliferative response of preparations of whole PBMC populations from 20 healthy individuals and 28 multiple sclerosis (MS) patients to purified protein derivative (PPD) and myelin basic protein (MBP) was monitored in a kinetic assay over a period of up to 10 days. PPD produced a classical secondary response in both groups, the magnitude being significantly reduced in the MS cohort. The magnitude and pattern of response to MBP did not differ between the two populations. The kinetic profile characteristic of a primary response was observed in both groups. Enrichment of the CD45RO+ve and CD45RA+ve T-cell subsets in PBMC led to a secondary response to PPD in the RO+ve and primary response in the RA+ve population in both groups. The response to MBP in both RO+ve and RA+ve populations exhibited primary kinetics in both MS patients and healthy individuals. However, the use of T-cell subset enriched populations allowed a finer dissection of the response to MBP which highlighted the more active role of RO-positive cells in MS patients. The most striking difference between patients and healthy individuals occurred on day 4 of culture when a greater response to MBP occurred in the CD45RO enriched population, paralleling the response to PPD, in the majority of patients. Futhermore in 4/8 patients and only 1/8 healthy individuals the response in the RO+ve cultures was maintained at a higher level than that seen in the corresponding RA+ve cultures throughout the culture period. This data indicates that a measurable memory response to MBP exists in MS patients implying prior activation of MBP reactive T lymphocytes during the course of disease. PMID- 11422211 TI - The role of C5a in the development of thrombotic glomerulonephritis in rats. AB - Thrombus formation is the important pathologic finding observed in glomerulonephritis induced by antiglomerular basement membrane (GBM) antibodies. Although strong deposition of C3 and membrane attack complex (MAC) is observed in this disease, the role of complement has not been fully elucidated. The aim of this work was to investigate the role of complement, especially an anaphylatoxin C5a, in a rat model of thrombotic glomerulonephritis. Rats were first pretreated with subclinical dose of lipopolysaccharide (LPS). Thrombotic glomerulonephritis was then induced by intravenous injection with rabbit antirat GBM (RbAGBM) (Group I). For the evaluation of the role of complement, the soluble complement receptor type 1 (sCR1) (Group II) or the C5a receptor antagonist peptide (C5aR-AP) (Group III) was intravenously administered 30 min before RbAGBM injection. For exploring the role of neutrophils, rats were pretreated with cyclophosphamide before induction of disease (Group IV). All rats were sacrificed at 6 h, and histological examination was performed. Rats in Group I developed severe glomerular thrombosis. Leucocyte accumulation and strong binding of C3 and MAC were observed in the glomeruli. In rats treated with sCR1 (Group II) and C5aR-AP (Group III), both leucocyte accumulation and thrombus formation in the glomeruli were significantly inhibited. C3 and MAC were negative in the glomeruli in Group II rats, while they were strongly observed in Group III. In neutrophil depleted rats (Group IV), there was also deposition of C3 and MAC in the glomeruli but thrombus formation was not observed. These findings indicated that glomerular thrombosis is dependent on the leucocytes, and mediated in part by the anaphylatoxin C5a but not MAC in the present model. PMID- 11422212 TI - Imbalance between interleukin-6 and adrenomedullin mRNA levels in peripheral blood mononuclear cells of patients with lupus nephritis. AB - In this study, we measured the mRNA levels of adrenomedullin (AM), C-type natriuretic peptide, vascular endothelial growth factor, interleukin-1beta (IL 1beta) and interleukin-6 (IL-6) in peripheral blood mononuclear cells (PBMC) of 34 patients with lupus nephritis (LN) (15 active and 19 inactive) and 30 healthy volunteers. mRNA levels were measured using a real-time quantitative PCR METHOD: Compared with healthy volunteers, IL-6 mRNA levels were elevated in LN patients (P < 0.005), while AM mRNA levels were decreased (P < 0.05). Also, IL-6 mRNA levels were higher and AM mRNA levels lower in active LN patients compared with inactive LN patients. In addition, IL-6 mRNA levels positively correlated and AM mRNA levels negatively correlated with SLE disease activity index and laboratory findings, such as blood urea nitrogen, serum creatinine, 50% haemolytic unit of complement and urinary excretion of protein over 24 h. Furthermore, IL-6 mRNA levels were negatively correlated with AM mRNA levels within the same LN patients. With regard to pathological findings, our results showed that IL-6 mRNA levels were higher, and AM mRNA levels significantly lower in patients with a high activity index compared to those with a low activity index. Following treatment with prednisolone, IL-6 mRNA levels in active LN patients decreased and AM mRNA levels increased to levels comparable to those in inactive LN and healthy volunteers. In vitro studies further demonstrated that elevated IL-6 mRNA levels in active LN patient PBMC were suppressed by the addition of adrenomedullin. Our results suggest that an imbalance between IL-6 and AM levels may play an important role in the progression of SLE, and that the mRNA levels of these genes in PBMC may be used as a disease activity index for SLE. PMID- 11422213 TI - N-acetylcysteine induces shedding of selectins from liver and intestine during orthotopic liver transplantation. AB - In orthotopic liver transplantation (OLT), N-acetylcysteine (NAC) reduces ischaemia/reperfusion (I/R) injury, improves liver synthesis function and prevents primary nonfunction of the graft. To further elucidate the mechanisms of these beneficial effects of NAC, we investigated influence of high-dose NAC therapy on the pattern of adhesion molecule release from liver and intestine during OLT. Nine patients receiving allograft OLT were treated with 150 mg NAC/kg during the first hour after reperfusion; 10 patients received the carrier only. One hour after reperfusion, samples of arterial, portal venous and hepatic venous plasma were taken and blood flow in the hepatic artery and the portal vein was measured. Absolute concentrations of sICAM-1, sVCAM-1, sP-selectin and sE selectin were not markedly different. However, balance calculations showed release of selectins from NAC-treated livers as opposed to net uptake in controls (P < or = 0.02 for sP-selectin). This shedding of selectins might be a contributing factor to the decrease in leucocyte adherence and improved haemodynamics found experimentally with NAC-treatment. PMID- 11422214 TI - Fibroblast growth factors in the developing central nervous system. AB - 1. It is now clear that members of the fibroblast growth factor (FGF) family have multiple roles during the formation of the central nervous system (CNS). 2. There are at least 23 members of the FGF family and, of these, 10 are expressed in the developing CNS, along with four FGF receptors (FGFR-1-4). 3. The present review discusses the roles of these FGFs, with emphasis on FGF-2, FGF-8, FGF-15 and FGF 17. Fibroblast growth factors-2 and -15 are generally expressed throughout the developing CNS, whereas FGF-8 and FGF-17 are tightly localized to specific regions of the developing brain and are only expressed in the embryo during the early phases of proliferation and neurogenesis. 4. Expression studies on FGFRs in the chick and mouse indicate that FGFR-1 is most generally expressed, whereas FGFR-2 and FGFR-3 show highly localized but changing patterns of expression throughout CNS development. The FGFR-4 has been localized to the developing CNS in fish but not at a detailed level, as yet, in chick or mouse. 5. A picture is emerging from these studies that particular FGFs signal through specific receptors in a highly localized manner to regulate the development of different regions of the brain. 6. This picture has been demonstrated so far for the developing cortex (FGF-2-/- mice), the forebrain and midbrain (FGF-8 hypomorphs) and the cerebellum (FGF-17/FGF-8 mutant mice). In addition, generation of mutant animals deleted for FGFR-1 and FGFR-2b IIIb demonstrate their importance in FGF signalling. 7. However, there are significant gaps in our knowledge of the localization of members of the FGF family and their receptors. More detailed information on the spatio-temporal mapping of FGFs and FGFR isoforms is required in order to understand the molecular mechanisms through which FGFs signal. PMID- 11422215 TI - Maternal environment alters renal response to angiotensin II in the spontaneously hypertensive rat. AB - 1. The level of hypertension displayed by the adult spontaneously hypertensive rat (SHR) may be reduced by 20-30 mmHg if SHR pups are cross-fostered to a normotensive dam at birth (SHRX). The mechanisms involved are largely unknown, but may involve the kidney and the renin-angiotensin system (RAS) because renal responses to angiotensin (Ang) II are enhanced in the SHR and brief blockade of the RAS in the young rat permanently lowers blood pressure in the SHR. Accordingly, the aim of the present study was to investigate the effect of cross fostering on the renal response to AngII in the SHR. 2. Renal function was studied in anaesthetized 4-week-old SHR, cross-fostered SHRX and control Wistar Kyoto (WKY) rats. Standard clearance methods were used to investigate the renal haemodynamic and tubular effects of AngII. The nitric oxide synthase (NOS) inhibitor NG-nitro-L-arginine methyl ester (L-NAME) was coinfused in some experiments to abolish the counterbalancing effect of nitric oxide. 3. Angiotensin II induced a fall in glomerular filtration rate (GFR) in both SHR and SHRX, but not in WKY rats. This was accompanied by a significant reduction in sodium excretion by SHR but not SHRX pups. This effect appeared to be tubular in origin, because sodium clearance was comparable between the two strains after AngII, but fractional sodium excretion was significantly higher in the SHRX. Coinfusion of the NOS inhibitor L-NAME had little further effect on sodium excretion in SHR and SHRX, but restored GFR in SHRX to levels comparable with those in WKY rats. 4. These data suggest that renal tubular sensitivity to AngII in the SHR can be altered during the early stages of postnatal development, which may contribute to the blood pressure- lowering effect of cross-fostering in the SHR. PMID- 11422216 TI - Ouabain-induced coronary vasoconstriction in cats is not neurally mediated. AB - 1. Previous reports indirectly implicate a neural mechanism for coronary constriction to centrally administered digitalis. However, autoregulatory changes in coronary resistance due to changes in arterial pressure may have influenced the interpretation of these studies. 2. We tested directly the hypothesis that cardiac sympathetic innervation is responsible for coronary constriction to ouabain by examining the effects of ouabain (intravenous (i.v.) and intracerebroventricular (i.c.v.)) before and after bilateral stellate ganglionectomy. 3. Cats were anaesthetized and instrumented for the measurement of heart rate, blood pressure and coronary blood flow velocity using an epicardial-attached suction Doppler probe. Animals were treated with atenolol and the effects of either i.v. or i.c.v. injections of ouabain were examined. 4. In seven cats treated with atenolol, i.v. ouabain (0.11 mg/kg) produced maximal increases in arterial pressure and coronary vascular resistance index (CVRI) of 66 +/- 7 mmHg and 37 +/- 9%, respectively. Following bilateral stellate ganglionectomy (n = 7), ouabain produced similar increases in arterial pressure (70 +/- 9 mmHg) and CVRI (39 +/- 7%). A higher dose of i.v. ouabain (1.1 mg/kg) produced maximal increases in arterial pressure (115 +/- 4 mmHg) and coronary resistance (86 +/- 14%) in intact cats (n = 6) that were similar to responses seen in cats in which stellate ganglionectomy had been performed (n = 6; arterial pressure 104 +/- 13 mmHg; coronary resistance 114 +/- 6%). The increases in coronary resistance to ouabain at both doses were significantly greater than increases in coronary resistance to passive elevation of arterial pressure during aortic constriction. Thus, autoregulation does not explain fully the coronary constriction to ouabain. 5. To further examine a central mechanism, i.c.v. perfusion with 0.3 mmol/L ouabain was performed in six cats, resulting in increases in arterial pressure (122 +/- 7 mmHg) and coronary resistance (58 +/- 14%). Similar increases in arterial pressure (117 +/- 16%) and coronary resistance (84 +/- 20%) were seen in separate studies (n = 6) following stellate ganglionectomy. 6. These results indicate that coronary constriction to ouabain does not require intact cardiac sympathetic innervation, but probably involves a direct or humorally mediated effect. PMID- 11422217 TI - Effect of metabolic control on nitrite and nitrate metabolism in type 2 diabetic patients. AB - 1. The measurement of nitrite and nitrate levels in plasma and urine is an approach to assess the activity of the endogenous nitric oxide (NO) system. The aim of the present study was to evaluate whether metabolic control may affect plasma levels and urinary excretion of nitrates and nitrites in type 2 diabetic patients. 2. Twenty consecutive type 2 diabetic patients were studied twice: first (study 1), under poor metabolic control; and, second, after improved metabolic conditions. Determinations of the main metabolic parameters and of plasma nitrates and nitrites (NOX) were performed in the fasting state. A 24 h urinary specimen was obtained for glycosuria, NOX and creatinine. Diet compliance and home blood glucose monitoring was evaluated on a weekly basis until study 2 was performed after 32 +/- 7 days: then, an identical protocol was repeated (study 2). 3. Fasting plasma glucose was lower in study 2 (8.27 +/- 2.11 vs 10.77 +/- 3.88 mmol/L, respectively; P < 0.05); similarly glycosylated haemoglobin (HbA1c) improved significantly. Plasma NOX levels were similar between the first and second studies (14.3 +/- 7.8 vs 13.5 +/- 8.1 micromol/L, respectively); nor were any differences observed in urinary NOX excretion rates (726 +/- 607 vs 689 +/- 444 micromol/day, respectively). The urinary excretion fraction of NOX was higher during study 1 than during study 2 (3.22 +/- 2.38 vs 1.88 +/- 1.98%, respectively; P = 0.031). A relationship was observed between fasting plasma glucose levels and the urinary excretion fraction of NOX (r2 = 0.12; P = 0.026). 4. In type 2 diabetic patients, plasma and urinary levels of NOX do not change after improvement of metabolic control. A worse metabolic control is associated with an increased urinary fraction excretion of NOX: thus, changes in plasma NOX concentration may reflect the effect of hyperglycaemia in the renal handling of these compounds rather than the effects on the L-arginine-NO pathway. PMID- 11422218 TI - Pirfenidone inhibits dimethylnitrosamine-induced hepatic fibrosis in rats. AB - 1. In the present study, we investigated the preventive effects of pirfenidone (PFD), an antifibrotic agent, on experimental hepatic fibrosis induced by dimethylnitrosamine (DMN) in rats. 2. Treatment with DMN caused a significant decrease in bodyweight and liver weight. Oral PFD (500 mg/kg daily for 4 weeks) essentially prevented this DMN-induced loss in bodyweight and tended to suppress the loss in liver weight. There were no significant differences in liver weight and serum L-alanine aminotransferase levels between PFD-treated and -untreated groups. Pirfenidone has no major side effects in vivo. 3. Pirfenidone suppressed the induction of hepatic fibrosis determined by histological evaluation and reduced hepatic hydroxyproline levels. Expression of mRNA for type I collagen and transforming growth factor-beta in the liver was also suppressed by PFD treatment. 4. Because hepatic stellate cells (HSC) are the major cellular source of extracellular matrix in hepatic fibrosis, we examined the effects of PFD on type I collagen production in vitro using rat primary HSC cultures. Pirfenidone inhibited collagen production in HSC culture in a dose-dependent manner. 5. These results demonstrate that the inhibitory effects of PFD against hepatic fibrosis may be due, at least in part, to blockade of collagen production by HSC and suggest that PFD may be potentially useful in the prevention of the development of hepatic fibrosis. PMID- 11422219 TI - Effects of L-arginine and furosemide on blood pressure and renal function in volume-expanded rats. AB - 1. The aim of the present study was to investigate the effects of L-arginine (L Arg) on blood pressure and water and electrolyte excretion in control and extracellular fluid volume-expanded rats (10% bodyweight with 0.9% NaCl) and to determine whether diuretic treatment with furosemide (FUR) can be optimized by the administration of L-Arg in this model. 2. Both groups of animals were anaesthetized, divided into groups and treated with either 7.5 mg/kg FUR, 250 mg/kg L-Arg, 1 mg/kg NG-nitro-L-arginine methyl ester (L-NAME), FUR + L-NAME or FUR + L-Arg. Mean arterial pressure (MAP), diuresis, natriuresis and kaliuresis were determined. 3. Extracellular fluid volume expansion induced no changes in MAP in control and volume-expanded rats (92+/-6 vs 100+/-8 mmHg, respectively). The hypotension induced by FUR in control and volume-expanded rats (69+/-7 and 76+/-5 mmHg, respectively) was significantly (P < 0.01) enhanced by the administration of L-Arg (54+/-3 and 64+/-3 mmHg, respectively). 4. Injection of L NAME increased MAP and diminished diuresis, natriuresis and kaliuresis in both groups. 5. Furosemide-induced water and electrolyte excretion was blunted by the administration of L-NAME. 6. The combination of L-Arg + FUR increased diuresis induced by FUR alone (control rats: 29.33+/-1.68 vs 12.91+/- 0.41 microL/min per 100 g, respectively; volume-expanded rats: 248.5+/-25.4 vs 112,6+/-8.3 microL/min per 100 g, respectively; P < 0.01). 7. The administration of the combination of L Arg + FUR promoted a decrease in the sodium/water excretion ratio compared with the administration of FUR alone (control rats: 0.230+/-0.018 vs 0.45+/-0.03, respectively, P < 0.001; volume-expanded rats: 0.091+/-0.010 vs 0.22+/-0.03, respectively, P < 0.01). 8. The potassium/water excretion rate induced by FUR alone and in the presence of L-Arg followed a pattern similar to that seen for natriuresis (control rats: 0.35+/-0.05 vs 0.20+/-0.05 microEq/min per 100 g, respectively; volume-expanded rats: 0.045+/-0.008 vs 0.014+/-0.003 microEq/min per 100 g, respectively; P < 0.01). 9. The decrease in the electrolyte/water excretion ratio observed with FUR + L-Arg in volume-expanded rats was greater than in control animals. 10. The results of the present study show that the administration of FUR with L-Arg contributes to enhanced hypotensive and diuretic effects of FUR, thus diminishing the relative electrolyte excretion in normal conditions and in extracellular fluid volume expansion. PMID- 11422220 TI - Airway protection following simulated gastro-oesophageal reflux in sedated and sleeping neonatal piglets during active sleep. AB - 1. In infants, promethazine has been implicated in the pathogenesis of sleep apnoea, apparent life threatening events (ALTE) and the Sudden Infant Death syndrome (SIDS). The aim of the present study was to investigate, in a neonatal animal, the effects of a commonly used promethazine-containing medication on airway protective mechanisms and cardiorespiratory reflexes following simulated gastro-oesophageal reflux (GER) to different levels in the oesophagus and pharynx. 2. Physiological and radiographic recordings were made in 21 naturally sleeping (controls) and 21 sedated (1.5 mg/kg, p.o., promethazine) piglets. On 3 consecutive days physiological recordings were made in all piglets during active sleep. Gastro-oesophageal reflux was simulated by the injection of boluses of 0.5 mL HCl, pH 2 or 3, or NaCl (0.9%) at 37 degrees C into the pharynx, upper or lower oesophagus. 3. In healthy neonatal piglets, minimal sedation with promethazine, which did not affect behaviour during wakefulness, revealed previously unreported findings during active sleep. 4. The most significant effects were observed following simulated GER to the pharynx, with no effect observed in the lower oesophagus. In sedated piglets, compared with naturally sleeping piglets, there was a significant reduction in swallowing (P < 0.01), delayed radiological clearance of fluid (P < 0.05), a reduction in breathing rate, oxygen saturation and heart rate and an increase in apnoea. 5. These findings are consistent with a low dose of promethazine producing a significant attenuation of airway protective mechanisms and, thus, stimulation of the laryngeal chemoreflex. The results suggest a mechanism for the association observed between promethazine use and the occurrence of ALTE and SIDS. The results support continued caution and suggest the need for greater regulation of promethazine-containing medications in infants. PMID- 11422221 TI - Mechanisms of endothelin-1-induced potentiation of noradrenaline response in rat mesenteric artery. AB - 1. Subthreshold concentrations of endothelin (ET)-1 enhance the contractile responses to noradrenaline (NA). We investigated possible mechanisms underlying the ET-1-induced enhancement of vasoconstrictor responses to NA in rat perfused mesenteric arteries. 2. Perfusion of arteries with subpressor dose of ET-1 (3 x 10-10 mol/L) significantly potentiated the pressor responses to NA (10-6, 3 x 10 6 and 10-5 mol/L) and this action of ET-1 was endothelium independent. 3. The protein kinase C (PKC) inhibitors staurosporine (10-8 mol/L) and calphostin C (10 7 mol/L) markedly attenuated the ET-1-induced enhancement of NA responses. Vasoconstrictor responses to NA were potentiated when vessels were perfused with phorbol 12-myristate 13-acetate (10-8 mol/L). 4. The potentiating effect of ET-1 was efficiently suppressed by Y-27632 (10-6 mol/L), a selective Rho-kinase inhibitor. In the presence of both staurosporine and Y-27632, contractile responses to NA alone were decreased markedly and ET-1-induced potentiation was abolished. 5. Both staurosporine and Y-27632 decreased contractile responses to NA in arteries of deoxycorticosterone acetate (DOCA)-salt hypertensive rats to levels observed in normotensive control animals. 6. These findings suggest that ET-1-mediated potentiation of responses to NA occurs through activation of either PKC or Rho-kinase. This mechanism seems to contribute to the enhanced vasoconstrictor responces to NA observed in DOCA-salt hypertensive rats, in which the responses to NA are enhanced tonically by endogenous vascular ET-1. PMID- 11422222 TI - Prophylactic effects of pilocarpine hydrochloride on xerostomia models induced by X-ray irradiation in rats. AB - 1. In the present study, we investigated the prophylactic effects of pilocarpine hydrochloride on xerostomia models induced by either single (15 Gy) or repeated (8.6 Gy x3 days) X-ray irradiation in rats. Pilocarpine hydrochloride was administered orally 90 min before each irradiation session. Then, 7 days later, salivary volume, amylase activity and protein concentration in the saliva secreted from the right parotid gland were measured before and after a subsequent administration of pilocarpine hydrochloride (intraduodenal). 2. In irradiated no pretreatment rats, irradiation induced a significant reduction in both spontaneous and pilocarpine hydrochloride-stimulated secretion (both total salivary volume and flow rate), regardless of the protocol used for X-ray exposure. In irradiated, pilocarpine hydrochloride-pretreated rats, salivary secretion was increased after stimulation by pilocarpine hydrochloride (intraduodenal) to a degree that depended on the pretreatment dose of pilocarpine hydrochloride (p.o.) in both xerostomia models. 3. There were no differences in amylase or protein concentrations between irradiated rats pretreated with pilocarpine hydrochloride and irradiated no-pretreatment control rats. 4. A decrease in the weight of the parotid gland was observed in rats exposed to either the single dose or repeated irradiation protocols. Changes in the submandibular gland were less marked than those in the parotid gland. These changes in gland weight were not affected by pilocarpine hydrochloride pretreatment. 5. The responsiveness of the parotid gland to subsequent stimulation with pilocarpine hydrochloride was apparently preserved in both xerostomia models by pretreatment with pilocarpine hydrochloride, which itself increased salivary secretion. This suggests that pilocarpine hydrochloride may exert functional protective effects against xerostomia that occurs following irradiation therapy through a stimulation of salivary secretion. PMID- 11422223 TI - The pathological consequences of anaemia. AB - Many patients with chronic diseases such as chronic renal failure, chronic inflammatory bowel disease and rheumatoid arthritis are anaemic. Recently congestive heart failure (CHF) has also been found to be associated with anaemia. In all these diseases this anaemia or chronic disease is at least partially due to excessive production of cytokines and leukotrines that interfere both with the effect of erythropoietin (EPO) at the bone marrow and the release of stored iron in the reticuloendothelial system. Treating this anaemia with subcutaneous EPO and IV iron improves the weakness, fatigue, cachexia, nutritional state, mood, cognitive function and quality of life. In the case of CHF it also improves cardiac function and patient functional class, prevents deterioration of renal function and markedly reduces hospitalization. Very few agents in medicine improve so many aspects of the patient so well and so quickly. Unfortunately (for the suffering patient) this anaemia is often ignored and goes untreated. PMID- 11422224 TI - Flow-cytometric analysis of erythrocytes and reticulocytes in congenital dyserythropoietic anaemia type II (CDA II): value in differential diagnosis with hereditary spherocytosis. AB - Congenital dyserythropoietic anaemia type II (CDA II) is the most common congenital dyserythropoietic anaemia. CDA II is frequently misdiagnosed as Hereditary Spherocytosis (HS) due to the presence of mild chronic haemolytic anaemia with splenomegaly, increased osmotic fragility, and presence of microspherocytes. Accurate diagnosis of CDA II is important to prevent severe iron overload. Erythrocyte and reticulocyte indices were assessed in 10 patients from six families with CDA II, 18 patients from eight families with HS, and 50 normal controls. Characteristic increases in distribution width were present in CDA II for cell volume (RDW, anisocytosis) and in HS for cell haemoglobin concentration (HDW, anisochromia), resulting in an RDW/HDW ratio which was significantly greater in CDA than HS (P < 0.0002). A cut-off value for RDW/HDW of 5.34 resulted in 89% sensitivity and 70% specificity in distinguishing CDA II from HS. Distribution width for cell haemoglobin content of reticulocytes (CHDWr) was characteristically increased in CDA II, resulting in a CHDW/CHDWr ratio significantly lower in CDA II than HS (P < 0.0002). A cut-off value of 0.98 provided 89% sensitivity and 80% specificity in distinguishing CDA II from HS. These differences in distribution widths of flow-cytometric parameters of reticulocytes and mature erythrocytes reflect the different pathogeneses of the two diseases and are helpful for the differential diagnosis of these two conditions. PMID- 11422225 TI - High serum folates and the simplification of red cell folate analysis. AB - Recent substantial increases in clinical blood folate concentrations are noted. Since red cell folates (RCF) are calculated from whole blood folates (WBF) by subtraction of the endogenous serum folate (SF) component, the reporting of clinical RCF results may be delayed because an ever increasing proportion (15%) of diagnostic SF levels are high (> 20 ng/ml) and need a repeat analysis. We evaluated 'plasma replacement' as a simple preanalytical procedure in which endogenous blood plasma is removed from red cells by washing and substituted with 'low-folate' plasma (serum) as an alternative conjugase (gamma-glutamyl carboxypeptidase) source for folate polyglutamate hydrolysis. Washed and conventional RCF assays compared well after both manual (n = 115, r = 0.98, y = 1x + 1.26) and automated washing of red cells (n = 170, r = 0.96, y = 0.96x - 0.73 ng/ml) and were not significantly different. The interassay reproducibility of folate results from washed blood samples was good (CV = < 6%). This novel 'plasma replacement' step halves the cost of a valid RCF assay by eliminating the need for endogenous SF analysis, and it expedites the reporting of clinical results. PMID- 11422226 TI - The feasibility of estimating the erythrocyte sedimentation rate within a few minutes by using a simple slide test. AB - The acute phase response is accompanied by the appearance of aggregated red blood cells in the peripheral blood. The Westergren erythrocyte sedimentation rate (ESR) is an indirect measurement of this enhanced aggregability. We adopted a simple slide test and image analysis to reveal the adhesiveness/aggregation of red blood cells. A significant correlation was found between the erythrocyte adhesiveness/aggregation test (EAAT) and the ESR. A predictive model for ESR based on EAAT and the age of the patients was created. This new approach will enable us to obtain within a few minutes a good estimate of whether a given individual has a mild moderate or significant acute phase response. With further development, we will be able to use a bedside small cartridge that will deliver the extrapolated ESR at low costs and within a couple of minutes. PMID- 11422227 TI - Patients with pulmonary and cardiac disease show an elevated proportion of immature reticulocytes. AB - The value of reticulocyte analysis has been largely confined to classification of anaemia. In the present study, we have investigated the value of undertaking automated fluorescent reticulocyte analysis (Abbott Cell-Dyn 4000) in patients with pulmonary or cardiac diseases. A control group of nonanaemic (n = 367) and anaemic patients (n = 57) was established and thereafter compared with a group of patients with pulmonary disease (172 without anaemia, 92 with anaemia) and another group with cardiac disease (520 without anaemia, 254 with anaemia). The Hb level and reticulocyte RNA content (as measured by the immature reticulocyte fraction, IRF) of the study subjects were inversely correlated (r = -0.41). The mean IRF of nonanaemic patients with pulmonary (mean = 0.331, SD=0.124) and cardiac disease (mean = 0.266, SD = 0.079) were both significantly higher (P < 0.05) than in control subjects (mean = 0.220, SD = 0.062). Nonanaemic and anaemic patients with cardiac disease both showed significantly higher (P < 0.05) reticulocyte percentage counts than the equivalent anaemic and nonanaemic controls. Each of these phenomena can all be attributed to hypoxia mediated EPO production. Our observation that 35% of patients with pulmonary disease have elevated levels of IRF, may prove useful in screening for tissue hypoxia in nonanaemic patients. Furthermore, serial monitoring of the IRF may prove valuable in observing the efficacy of therapy in these patients. PMID- 11422228 TI - The analysis of eosinophil and lymphocyte phenotype following single dose of high dose methylprednisolone in two siblings with marked hypereosinophilia. AB - The effect of single high-dose methylprednisolone (HDMP) on eosinophil and lymphocyte phenotype in two siblings with marked hypereosinophilia was investigated. Since conventional dose steroids failed to produce a haematological and clinical response in case one, both children were given HDMP (20 mg/kg/day). Eosinophils and lymphocytes showed an activated state before treatment, characterized by marked expression of CD11b, CD18, CD45RO on eosinophils and increased HLA-DR, CD95, CD18, CD38 on lymphocytes. Twenty-four hours after administration of HDMP a dramatic reduction in peripheral blood eosinophil count was observed in both patients associated with phenotypic changes characterized by decreased expression of CD11b, CD18, CD13 and increased CD95 expression in one. Furthermore, HDMP treatment induced a drop in the expression of CD95 and CD18 on lymphocytes. These changes may suggest a suppressive role for HDMP on eosinophil and lymphocyte activation which may have contributed to the haematological and clinical response. PMID- 11422229 TI - Quinolone resistance in neutropenic patients: the effect of prescribing policy in the UK and Pakistan. AB - Quinolones are increasingly used as prophylaxis in neutropenic patients to prevent serious Gram-negative septicaemias but practice is not uniform because of the controversial evidence as to their effectiveness. It is unclear if they are of real benefit in patients with short episodes of neutropenia such as those resulting from treatment for solid tumours and lymphomas. The concern over the use of ciprofloxacin in such settings is the increasing development of quinolone resistant Gram-negative bacteria. We have retrospectively analysed our bacterial isolate resistance patterns in the Queen Elizabeth Hospital (QE) and in the Haematology Department of the Aga Khan Hospital (AKU), Pakistan where all patients would receive ciprofloxacin prophylaxis when neutropenic. Seven out of 57 (12.2%) and 18 out of 55 (32.7%) Gram-negative organisms isolated from blood cultures at the QE and AKU Haematology Departments, respectively, were resistant to ciprofloxacin (P < 0.01). In the Birmingham community this was significantly lower (P < 0.01) (55 out of 6423: 0.85%). We also showed a higher level of E. coli resistance at the AKU (18 out of 31: 58%) where ciprofloxacin use was more widespread than at the QE (1 out of 11, P < 0.01). We conclude that ciprofloxacin should not be used indiscriminately. PMID- 11422230 TI - Low level leucocyte counting: a critical variable in the validation of leucodepleted blood transfusion components as highlighted by an external quality assessment study. AB - Leucocyte counts of < 5 x 106 per blood transfusion product are currently recommended in the UK in order to reduce transfusion-related infections and febrile reactions. Routine leucocyte depletion, however, requires the development of reliable internal and external quality assurance (EQA) programmes. We report preliminary findings from the UK NEQAS for Low-Level Leucocyte Counting from 18 UK Transfusion Centres over a four month period. Data analysis showed that the IMAGN 2000 had the lowest CVs (range 7.5-36%, mean 16.7) for samples with counts of 5-30 cells/microl when compared to the flow cytometric (range 13.8-88%, mean 29.5) and Nageotte methods (range 20.6-117%, mean 61.8). In addition, laboratories using commercial nuclear stains (LeucoCOUNTTM) had consistently lower CVs than those using 'in-house' propidium iodide staining methods. Important differences in flow cytometric gating strategies were also identified. This study highlights the current variability in low level leucocyte counting, especially within the critical range of 5-30 cells/microl (equating to < 5 x 106/l). The acceptance of consensus protocols, including gating strategies and nuclear staining techniques, is required to reduce the observed interlaboratory variation. Finally, we demonstrate that stabilized blood preparations can be successfully used to provide a national/international low-level leucocyte EQA scheme. PMID- 11422231 TI - Haemoglobin Q-Thailand and hereditary spherocytosis in a Chinese family. AB - A Chinese family with concurrent hereditary spherocytosis (HS) and haemoglobin (Hb) Q-Thailand is described. The Hb Q-Thailand mutation was found on the remaining alpha1 globin gene on a chromosome 16 containing the (-alpha 4.2) deletion. Active haemolysis in members of this family is segregated with the HS phenotype, and the Hb Q-Thailand in the heterozygous state does not seem to show any modulating effect on HS. PMID- 11422232 TI - Otological manifestations of acute leukaemia: report of two cases and review of literature. AB - Otological manifestations exceptionally reveal acute leukaemia, whereas leukaemic infiltration of any tissue is frequent at postmortem examination. We present two cases of acute myeloblastic leukaemia revealed by a loss of hearing due to a middle-ear leukaemic infiltration. The characteristics of such a clinical and radiological finding are emphasized. It is suggested that middle and inner ear can be a 'sanctuary' localization, which might sometimes require radiation therapy to achieve durable and complete remission. PMID- 11422233 TI - Prothrombin time inhibition detected with recombinant but not with conventional thromboplastins in two patients with high-titre IgM and moderate-titre IgA anticardiolipin antibodies. AB - We report two cases of high-titre IgM and moderate-titre IgA anticardiolipin antibodies (ACA) in whom prothrombin times (PT) derived using recombinant thromboplastins (rTP) were prolonged but were normal when measured with conventional thromboplastins. The anticoagulant nature of these antibodies cannot be categorized as the classical lupus type. We suggest that routine screening for the presence of antiphospholipid antibodies (APA) should be performed in patients who fall into this category. PMID- 11422234 TI - Platelet apheresis for digital gangrene due to thrombocytosis in chronic myeloid leukaemia. AB - Thrombocytosis is a frequent presenting feature of myeloproliferate disorders and is associated with increased incidence of thrombotic and haemorrhage complications. However, these complications are rare in chronic myeloid leukaemia (CML). We describe a case of CML which presented with digital gangrene due to thrombocytosis. Reduction of the platelet count by plateletpheresis lead to rapid symptomatic relief and recovery from the gangrene. PMID- 11422235 TI - Metastatic transitional cell carcinoma or nonsecretory multiple myeloma? PMID- 11422236 TI - Purpuric rash following treatment with 2-chlorodeoxyadenosine. PMID- 11422237 TI - Polio eradication and after. PMID- 11422238 TI - European guidelines for urinalysis: a collaborative document produced by European clinical microbiologists and clinical chemists under ECLM in collaboration with ESCMID. PMID- 11422239 TI - Molecular-based strategies for assessment of CMV infection and disease in immunosuppressed transplant recipients. AB - OBJECTIVES: Molecular assays are now considered to be the "gold standard" for assessment of human cytomegalovirus (CMV) infection and disease in those at risk from severe associated clinical manifestations. There is, however little consistency in the methods used in different centres. This study was undertaken to compare different qualitative molecular-based approaches for assessment of CMV activation from latency in samples from immunosuppressed transplant recipients. METHODS: Nucleic acid amplification techniques based on the polymerase chain reaction (PCR) and nucleic acid sequence-based amplification (NASBA) were undertaken for the assessment of CMV replication and associated disease in immunosuppressed transplant recipients. Samples from 32 transplant recipients were tested during this study using three molecular-based strategies: (1) detection of CMV DNA in whole blood extracts (positive after a single round of PCR considered "high-level" positive, N = 55); (2) detection of cell-free CMV DNA in plasma (two methods, N = 55 for each); and (3) detection of late pp67 CMV mRNA after NASBA (N = 51). Results using a commercial pp65 antigenemia assay were available for comparison from 40 samples. RESULTS: Seven samples were positive for CMV by all methods and 36 were negative by all methods undertaken. The other 12 samples gave discordant results using different molecular methods. The correlation between whole blood "high-level" PCR, NASBA for pp67 mRNA and antigenemia results was generally good. Results presented show that plasma PCR results do not always correlate with methods utilizing whole blood as the substrate and that inhibitors in these samples could be problematic. Whole blood PCR gave more positive results than the other assays but use of a nested assay on whole blood or plasma led to detection of CMV in individuals who had no other indicators of virus replication and who did not develop associated disease (low specificity). Although the number of confirmed CMV disease episodes was low in this study, the problems of low positive predictive value for sensitive, qualitative PCR assays was clearly demonstrated. CONCLUSION: Assays based on qualitative detection of viral nucleic acid may provide information useful for management of CMV but caution is necessary when making comparisons between results using different molecular strategies. It remains to be proven in large, comparative clinical studies in which the approach and method give the best balance between sensitivity, specificity and clinical relevance for different patient groups. PMID- 11422240 TI - Polycolonization of Helicobacter pylori among Chinese subjects. AB - OBJECTIVE: To investigate the situation among Chinese patients with regard to infection with multiple strains of Helicobacter pylori. METHODS: Biopsy specimens for culture of H. pylori were obtained from gastric antrum, body and fundus of 20 patients during endoscopic investigation of upper gastrointestinal symptoms. H. pylori was identified by culture from one site in 16 and two or more sites in 10 of the 16 patients. Five isolated colonies of six strains of H. pylori from gastric antrum were subcultured and used for further analysis. Antibiotic susceptibility to metronidazole and clarithromycin was determined by disk diffusion test. Protein profiles of isolates were compared by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). DNA diversity of the isolates was determined by arbitrarily primed polymerase chain reaction (AP PCR) fingerprinting. RESULTS: Of the 10 patients with multiple isolates, 70% (7/10) exhibited variation in susceptibility to metronidazole and 20% (2/10) to clarithromycin between different sites. In 83% of (5/6) single colonies, no variability was seen in metronidazole and clarithromycin susceptibility; they were either susceptible or resistant. Protein profiles of all isolates by SDS PAGE were similar. Isolates from different patients produced clearly different AP PCR fingerprints. In 50% of H. pylori strains isolated from different sites of the stomach, genetic diversity was demonstrated by different AP-PCR fingerprints. In 67% (4/6) strains, five single-colony fingerprints were similar. CONCLUSIONS: Genetic variability has been found in H. pylori strains. Individual patients are infected with a single predominant genotype at a single site but can be colonized by multiple strains, and they may show different antibiotic susceptibilities. Individual colonies of the H. pylori population from a single site may not always yield identical DNA fingerprints and antibiotic sensitivities. PMID- 11422241 TI - Phenotypic and genotypic markers of Staphylococcus epidermidis virulence. AB - OBJECTIVES: To analyze Staphylococcus epidermidis strains, previously tested for their virulence in a mouse model of subcutaneous infection, for various phenotypic traits (biofilm density, extracellular polysaccharide, slime associated antigen (SAA)) and for the presence of the ica gene cluster, to determine which of these phenotypic and genotypic methods best correlates with virulence in the mouse model. METHODS: The quantitative biofilm assay was performed on 10 strains of S. epidermidis, comprising (1) RP62A (ATCC 35984), (2) the strongest and weakest biofilm producers in our collection, (3) a pair of phenotypic variants, and (4) a strain whose biofilm density was enhanced in iron limited media. Biofilm density was measured after growth at 37 degrees C and at ambient temperature, in trypticase soy broth (TSB) with and without glucose supplementation and using both chemical and heat fixation. Strains were assayed for SAA using a double immunodiffusion method. Extracellular polysaccharide was detected by transmission electron microscopy (TEM). A 546-base-pair segment of the ica gene cluster was amplified by PCR. RESULTS: Biofilm formation in TSB, glucose-enriched TSB, extracellular polysaccharide (observed by TEM), expression of SAA and presence of the ica gene predicted virulence of nine, nine, nine, eight and eight of 10 strains, respectively. The phenotypic expression of biofilm and related properties was medium and temperature dependent. We encountered one ica-positive strain that failed to express biofilm in standard TSB at 37 degrees C, but was virulent in a mouse model, and another strain that lacked ica, produced biofilm and was virulent in the model. CONCLUSIONS: Mouse virulence in our model can be predicted by any of the phenotypic or genotypic methods examined for > or = 80% of strains. Medium and incubation conditions affect the expression of phenotypic markers by some strains. For the remaining strains, possible reasons for inconsistencies between the presence of the ica gene, phenotypic markers and mouse virulence include (1) dependence of biofilm on genes other than ica, (2) sequence differences in ica, (3) dependence of biofilm expression in vivo on strain characteristics and media used to prepare inocula for in vivo studies. PMID- 11422242 TI - Evaluation of recomWell Treponema, a novel recombinant antigen-based enzyme linked immunosorbent assay for the diagnosis of syphilis. AB - OBJECTIVE: To evaluate the diagnostic performance of an enzyme immunosorbent assay (recomWell Treponema) for the diagnosis of syphilis. The novel recombinant antigens Tpn47, TpN17 and TpN15 were utilized. METHODS: A total of 782 human serum specimens, belonging to four different categories (blood donors, n = 200; routine laboratory screening for syphilis, n = 400; syphilis patients, n = 122; potential cross-reactors, n = 60), were evaluated to compare the sensitivity and specificity of the recomWell Treponema kit with a standard whole Treponema pallidum cell lysate antigen-based ELISA (Syphilis Screening) and with micro haemagglutination (MHA-TP). RESULTS: The overall specificity and sensitivity of the recomWell Treponema IgG was 98.9% and 98.3%, respectively. The specificity and sensitivity of Syphilis Screening ELISA was 98.7% and 98.3%, respectively. The agreement between recomWell Treponema and Syphilis Screening was 100%, 97.8%, 95.9% and 95% among the blood donor specimens, screening samples, syphilis specimens and the potential cross-reactors, respectively. Values of concordance varying from 96.7% to 98.3% were found in the different groups of sera between recomWell Treponema and MHA-TP. In addition, recomWell Treponema demonstrated a good diagnostic performance when used to detect the IgM to T. pallidum. No false positive sera were identified and, in 17/19 samples from primary infection, an IgM immune response was found. CONCLUSIONS: recomWell Treponema was shown to be a highly specific and sensitive method in all stages of syphilis screening and it can be considered as alternative to other ELISA tests based on native antigen preparations. PMID- 11422243 TI - Lack of antimicrobial activity of sodium heparin for treating experimental catheter-related infection due to Staphylococcus aureus using the antibiotic-lock technique. AB - OBJECTIVE: To elucidate the potential antimicrobial activity of sodium heparin in the treatment of catheter-infection using the antibiotic-lock technique. METHODS: We performed in vitro studies of the antibiotic susceptibility, stability and synergy of sodium heparin, vancomycin and ciprofloxacin. Efficacy studies were performed in a new animal model of Staphylococcus aureus catheter-related infection in which infection was produced via the endoluminal route. White New Zealand rabbits were surgically implanted with a sylastic catheter into the inferior cava vein. Immediately afterwards, infection was induced by filling and locking the catheters with 0.7 mL of broth culture containing 108 colony-forming units of S. aureus. Eighteen hours later the antibiotic-lock technique was started. Treatment groups were: control without treatment, sodium heparin at 2500 IU/mL, vancomycin at 2500 mg/L, ciprofloxacin at 1000 mg/L, vancomycin plus heparin and ciprofloxacin plus heparin. RESULTS: Sodium heparin showed an MIC90 higher than 6000 UI/mL against S. aureus causing catheter infection. Studies of antimicrobial synergy by the time-kill method between vancomycin and ciprofloxacin at MIC with sodium heparin at 2500 IU/mL showed no interactions. Vancomycin (2000 microg/mL) and ciprofloxacin (1000 microg/mL) in a solution containing sodium heparin (2500 IU/mL) were stable at 37 degrees C for a 72-h period. Two sets of in vivo experiments were carried out using differents strains of S. aureus. In both cases, sodium heparin showed no therapeutic efficacy when compared to control group and did not increase the antibiotic efficacy when used in combination with vancomycin or ciprofloxacin. CONCLUSION: Sodium heparin lacked antibacterial activity against S. aureus causing catheter-related infections. PMID- 11422244 TI - Microbiological characteristics of subgingival microbiota in adult periodontitis, localized juvenile periodontitis and rapidly progressive periodontitis subjects. AB - OBJECTIVE: To describe the prevalence of the cultivable subgingival microbiota in periodontal diseases and to draw attention to the polymicrobial nature of periodontic infections. METHODS: The study population consisted of 95 patients, 51 females and 44 males, aged 14-62 years. Twenty-nine patients exhibited adult periodontitis (AP), six localized juvenile periodontitis (LJP), and 60 rapidly progressive periodontitis (RPP). Two to four pooled bacterial samples were obtained from each patient. Samples were collected with sterile paper points from the deepest periodontal pockets. The samples were cultured under anaerobic and microaerophilic conditions using selective and non-selective media. Isolates were characterized to species level by conventional biochemical tests and by a commercial rapid test system. RESULTS: Prevotella intermedia and Capnocytophaga spp. were the most frequently detected microorganisms in all diagnostic groups. Porphyromonas gingivalis and Peptostreptococcus micros were found more frequently in AP and RPP patients, while Actinobacillus actinomycetemcomitans and Eikenella corrodens were associated with AP, LJP and RPP patients. The other bacterial species, including Actinomyces spp., Streptococcus spp. and Eubacterium spp., were detected at different levels in the three disease groups. CONCLUSIONS: The data show the complexity of the subgingival microbiota associated with different periodontal disease groups, indicating that the detection frequency and levels of recovery of some periodontal pathogens are different in teeth affected by different forms of periodontal disease. PMID- 11422245 TI - In vitro activity of cefpirome and vancomycin in combination against gentamicin susceptible and gentamicin-resistant Staphylococcus aureus. PMID- 11422246 TI - Biochemical characterization of chromosomal cephalosporinases from isolates belonging to the Acinetobacter baumannii complex. PMID- 11422247 TI - Detection of antibodies to Candida albicans germ tubes in heroin addicts with systemic candidiasis. PMID- 11422248 TI - 'Good laboratory practice' in diagnostic laboratories using nucleic acid amplification methods. PMID- 11422249 TI - Ecological effects on the oro- and nasopharyngeal microflora in children after treatment of acute otitis media with cefuroxime axetil or amoxycillin-clavulanate as suspensions. AB - OBJECTIVE: To evaluate if the extent of normal microflora disturbances differed between treatment with amoxycillin-clavulanate administered in an active form and cefuroxime axetil administered as an inactive prodrug. METHODS: Twenty-eight children, 0.5-5 years old, diagnosed with acute otitis media (AOM), were treated with either amoxycillin-clavulanate (13.3 mg/kg 3 times daily) or cefuroxime axetil (15 mg/kg twice daily) for 7 days. Saliva samples and nasopharyngeal swabs were collected before, directly after and 2 weeks after treatment. The saliva samples were quantitatively and qualitatively analyzed and the nasopharyngeal swabs were qualitatively analyzed. All isolated strains were tested for beta lactamase production. RESULTS: Both treatment regimens gave rise to similar alterations of the normal oropharyngeal microflora. In both groups, the amount of Streptococcus salivarius was significantly reduced (P < 0.05). The most common causative pathogens of acute otitis were S. pneumoniae, Haemophilus influenzae and Moraxella catarrhalis. On the day of enrollment, approximately half of the patients, in both groups, were infected with more than one pathogen. The rate of infection or colonization with more than one potential pathogen was low on day 7 but recurred 2 weeks after treatment to similar levels as on day 0. The total number of patients with reinfection, recolonization or recurrence of pathogens on day 21 was 11/12 in the amoxycillin-clavulanate group and 4/7 in the cefuroxime axetil group. The most common beta-lactamase producer was M. catarrhalis. CONCLUSION: The local high concentration of antibiotics in the oropharynx immediately after intake of antibiotic suspensions seem to have little or no impact on the extent of disturbance of the microflora in this region. Children of this age group seem prone to either reinfection, recolonization or persistence of pathogens within 2 weeks after treatment. Furthermore, co-infection with more than one pathogen seems common in children with AOM and infection with beta lactamase producing microorganisms occurs frequently. PMID- 11422250 TI - Typing of Pseudomonas aeruginosa strains in Norwegian cystic fibrosis patients. AB - OBJECTIVES: Typing of Pseudomonas aeruginosa isolates from Norwegian cystic fibrosis (CF) patients with chronic Pseudomonas lung infection in order to see whether cross-infection might have occurred. METHODS: Isolates from 60 patients were collected during the years 1994-98, and typed by pulsed field gel electrophoresis. RESULTS: Seventy-one strains were identified. One large cluster of identical strains included 27 patients, and 13 smaller clusters of 2-4 patients were found (26 patients). Seven patients had a strain not shared by other patients (private strains). Harboring the main cluster strain was significantly associated with participation in summer camps and training courses (P = 0.004, chi-squared test). There were no associations with regular admissions to hospital (intravenous antibiotic courses) or smaller social gatherings of short duration. Small clusters and private strains were not associated with any of the risk factors. All strains were sensitive to colistin. The minimal inhibitory concentrations were generally lower in Norwegian P. aeruginosa strains compared with isolates from Danish patients. CONCLUSIONS: Our results indicate that cross-infection with P. aeruginosa between cystic fibrosis patients has occurred. PMID- 11422251 TI - Amplified fragment length polymorphism genotyping of metronidazole-resistant Helicobacter pylori infecting dyspeptics in England. AB - OBJECTIVE: Intra-specific diversity of Helicobacter pylori infecting stomachs of different individuals was investigated by numerical analysis of amplified fragment length polymorphisms (AFLP), to determine the existence of clones within the strain population and the effect that antibiotic treatment, particularly with metronidazole (Mtz), had on the balance of types/subtypes present before and after treatment. METHODS: The 92 cultures studied comprised 89 single or multiple (pre- and post-treatment) isolates from gastric biopsies from 35 dyspeptic patients at two geographical locations in England, and three reference strains. HindIII restriction fragments tagged with specific adaptors were used as template DNA for AFLP. Patterns were coded in binary format according to deduced sizes of amplified fragments, and numerical analysis was performed. RESULTS: H. pylori isolated from different individuals were highly diverse (43 AFLP types) with a continuum of similarities that included three putative strain clusters at the 55% similarity level. Twelve sets each comprised identical isolates but subclonal variants with similarities of 82-99% coexisted in isolate sets from 19 patients. Seven sets contained strains with different AFLP types which for several corresponded with vacA/cagA genotypic differences. Mtz resistance was a feature of clonal as well as unrelated isolates. CONCLUSIONS: AFLP profiling was a robust, reproducible and highly discriminatory means of indexing H. pylori strain diversity, and the numerical analysis enabled clonal/subclonal variants infecting an individual to be defined and contrasted with the general species diversity. The majority (65%) of patients had co-infections with different strain types/subtypes but antibiotic treatment apparently did not markedly modify H. pylori population diversity in individual stomachs. Mtz sensitivity was generally associated with greater strain diversity as several subtypes often coexisted in sensitive pretreatment strain sets. In contrast, Mtz-resistant strain populations were less diverse, which was attributed to selection by previous exposure to nitroimidazoles in the same or a different host. PMID- 11422252 TI - Detection of human cytomegalovirus (HCMV) pp67-mRNA and pp65 antigenemia in relation to development of clinical HCMV disease in renal transplant recipients. AB - OBJECTIVE: To evaluate the performance of the recently introduced method based on detection of human cytomegalovirus (HCMV) pp67 mRNA in blood by the nucleic acid sequence-based amplification (NucliSens), in comparison to semiquantitative detection of pp65 HCMV antigen in white blood cells, in relation to development of clinical HCMV disease. METHODS: Thirty patients, recipients of renal transplants, were monitored prospectively for the presence of pp67 mRNA, the presence and level of pp65 antigenemia, IgG and IgM antibodies, and the development of clinical HCMV disease. A total of 148 samples were examined during the observation period. RESULTS: Twenty-five samples were positive for pp67-mRNA and 45 samples contained at least one pp65 positive cell, with 68% agreement between the two assays. Both assays predicted correctly the development of clinical disease in five patients, giving a sensitivity of 100%. However, the specificity of the pp67-mRNA test was 72%, and of the pp65 antigenemia test from 20 to 64%, depending on the level of antigenemia chosen for cut-off. pp67-RNA appeared somewhat earlier than pp65 antigenemia, and responded earlier to treatment. Sero-conversion and appearance of IgM antibodies were of very little clinical value. CONCLUSION: Both the pp67-mRNA and the pp65 antigenemia assay predicted correctly the development of clinical HCMV disease in renal transplant recipients. However, the specificity of both tests with respect to development of HCMV disease, especially the pp65 antigen test was moderate. Significantly positive tests not necessarily prove the development of clinical disease. Testing for pp67-mRNA may improve the diagnosis and management of HCMV disease in renal transplant patients. PMID- 11422253 TI - Cryptosporidiosis in Kuwaiti children: seasonality and endemicity. AB - OBJECTIVE: To detect the incidence of cryptosporidial infection in children presenting with gastrointestinal symptoms at the local hospital in Kuwait. METHODS: Over a period of 3 years, September 1995 to August 1997, a single fecal sample from 3549 children was analyzed by modified saframin-methylene blue (SM-B) staining and a direct immunofluorescence test for the presence of Cryptosporidium oocysts. RESULTS: Cryptosporidium oocysts were detected in 51 (10%) children with diarrhea. Prevalence was highest (73%) in children > 2 years of age compared with children < 2 years of age. The maximum number of cases, 38 of 51 (75%), was seen during the months January to April, indicating a marked seasonal variation. Polyparasitism was common in children with diarrhea; however, 43 of the 51 (84%) children with cryptosporidiosis had cryptosporidium infection only. Blastocystis hominis and Endolimax nana were the most common parasites detected (38% and 15%, respectively). Forty-seven of the 51 (90%) children with cryptosporidiosis were Kuwaiti and gave no history of travel abroad, suggesting that the infection was acquired indigenously. Sociodemographic information on children with cryptosporidiosis suggests three possible modes of transmission of infection: drinking contaminated water stored in overhead water tanks, person to person, or contact with infected animals. CONCLUSION: In this study, we observed water-borne transmission of cryptosporidium infection in children with diarrhea. The infection is seasonal and endemic. PMID- 11422254 TI - Incidence of Staphylococcus aureus with reduced susceptibility to glycopeptides in two French hospitals. AB - OBJECTIVES: To determine the incidence of Staphylococcus aureus isolates with reduced susceptibility to glycopeptides among all clinical isolates collected consecutively in two French hospitals between November 1998 and April 1999. METHODS: Methicillin-resistant and -susceptible S. aureus isolates were screened on vancomycin- or teicoplanin-supplemented agar plates. Glycopeptide MICs were determined by the E test procedure with a high inoculum and by an agar dilution technique. Glycopeptide-intermediate S. aureus isolates were identified as homogeneously or heterogeneously resistant to vancomycin by performing population analysis. RESULTS: Of the 640 isolates recovered from 518 patients, three from the same patient and two from two different patients showed homogeneous or heterogeneous intermediate resistance to vancomycin. CONCLUSION: The incidence of glycopeptide-intermediate S. aureus (homogeneously or heterogeneously resistant) in a non-selected patient population, i.e. regardless of predisposing factors and glycopeptide therapeutics, remains low in the two French hospitals involved in the study, representing 0.6% of isolates. PMID- 11422255 TI - Clinical microbiological case: thermointolerant microorganism growth in blood cultures and catheter tip. PMID- 11422256 TI - Endocarditis due to Neisseria mucosa after tongue piercing. PMID- 11422257 TI - Acute pulmonary hypertension following paclitaxel in a patient with AIDS-related primary effusion lymphoma. PMID- 11422258 TI - Prevalence of extended-spectrum beta-lactamases in group-1 beta-lactamase producing isolates. PMID- 11422259 TI - Linezolid breakpoints. PMID- 11422260 TI - Validity and ethics of the human 4-h patch test as an alternative method to assess acute skin irritation potential. AB - For more than 50 years, the Draize rabbit skin irritation test has reigned supreme as the regulatory method of choice for the identification of skin irritant chemicals. To date no in vitro alternative test has been validated as an adequate replacement. However, one potential option, to test the endpoint of concern (skin irritation) in the species of concern (man) has been overlooked. The advent of predictive in vitro tools for the identification of substances corrosive to the skin has opened up the practical possibility of carrying out safe and ethical studies on small panels of humans. The human 4-h patch test has been developed to meet the needs of identifying chemical skin irritation potential, providing data which is inherently superior to that given by a surrogate model, such as the rabbit. This paper reviews in detail the present state of the human 4-h patch test, highlighting its advantages and noting its utility as the 'gold standard' on which to build future in vitro models. PMID- 11422261 TI - Contact dermatitis in Korean dental technicians. AB - The high risk of occupational contact dermatitis in dental personnel are well accepted throughout the world. There are few reports concerning occupational skin disease in dental personnel in Korea. The purposes of this study were to investigate the frequency, characteristics and causative factors of contact dermatitis in Korean dental technicians. Recording of personal history, physical examination and patch tests with the Korean standard series and dental screening series were performed in 49 dental technicians. Most of the subjects were exposed to a variety of compounds, including acrylics, metals, plaster, alginate, etc. 22 (44.9%) subjects had contact dermatitis, present or past, and the site involved was the hand in all 22. The most common clinical feature of hand dermatitis was itching (77.3%); scaling, fissuring and erythema were other common clinical features. Metals, including potassium dichromate (24.5%), nickel sulfate (18.4%), mercury ammonium chloride (16.3%), cobalt chloride (12.2%) and palladium chloride (10.2%), showed high positive rates in patch test results of 49 dental technicians. 7 positive reactions to the various acrylics were found in 3 subjects. In our study, the frequency and clinical features of the contact dermatitis showed a similarity to other reports, though the patch test results were somewhat different; a higher patch-positive reaction to metals and a relatively lower patch-positive reaction to acrylics than the patch test results reported in Europe. PMID- 11422262 TI - Occupational contact dermatitis has an appreciable impact on quality of life. AB - In this study, we examined the impact of occupational contact dermatitis on quality of life. 181 patients, diagnosed with occupational contact dermatitis over a period of 3 years (1996-1999 inclusive), were sent a questionnaire based on the Dermatology Life Quality Index (DLQI) and the Short Form-36 (SF-36). 60 (32%) patients were in industrial occupations and 27 (14%) in health care. An overall response rate of 39% (n=70) was obtained. The median DLQI score was 5, with a mean score 6.6 (SD 6.4), which is similar to that seen in Behcet's syndrome and urticaria. There was no statistically-significant difference between male and female median scores (p=0.98) and no significant correlation between age and DLQI score nor between DLQI score and time from diagnosis. The most problematic quality of life areas were symptoms and feelings. Males scored highest in problems associated with work, relationships and treatment, whereas females scored highest in problems associated with symptoms and feelings, daily activities and leisure. The SF-36 scores showed an association between physical problems and emotional problems affecting work. From this study, it can be seen that occupational contact dermatitis has an appreciable impact on quality of life. PMID- 11422263 TI - Standardization of the TRUE Test imidazolidinyl urea and diazolidinyl urea patches. AB - The preservatives imidazolidinyl urea (IMID, Germall 115) and diazolidinyl urea (DU, Germall II) are commonly used in cosmetic products and are well-known sensitizers. The aim of the present study was to establish the optimal patch test concentration in hydrophilic dried-in vehicle (TRUE Test) for IMID and DU. 181 patients were included in the study. Of these, 150 were patients referred for patch testing, 12 were patients with known allergy to IMID and 19 were patients with known allergy to DU. 76 consecutive patients and the 12 IMID-allergic patients were patch tested with a dilution series IMID (0 to 600 microg/cm(2)), formaldehyde (180 microg/cm(2)) and DU (200 microg/cm(2)). 74 consecutive patients and the 19 DU-allergic patients were patch tested with a dilution series of DU (0 to 600 microg/cm(2)), formaldehyde (180 microg/cm) and IMID (200 microg/cm(2)). A positive dose-response relationship was found. The number of doubtful reactions decreased with increasing test concentrations. No late reactions were observed. A patch test concentration in hydrophilic dried-in vehicle (TRUE Test) of 600 microg/cm(2) was found to be adequate and safe for both IMID and DU. PMID- 11422264 TI - Diluted Compositae mix versus sesquiterpene lactone mix as a screening agent for Compositae dermatitis: a multicentre study. AB - The objective of this study was to compare diluted Compositae mix with sesquiterpene lactone mix as a marker for Compositae dermatitis. Patients undergoing diagnostic patch testing in 8 British Isles patch test centres were also patch tested to sesquiterpene lactone mix (0.1% pet.) and Compositae mix (3% pet. or 0.6% pet.). Of a total of 5574 patients tested, 71 (1.3%) reacted to Compositae mix, 57 (1.0%) reacted to sesquiterpene lactone mix, and 45 to both. Diluted Compositae mix remained a more sensitive screening marker for Compositae allergy. We recommend that patients are patch tested to both agents. PMID- 11422265 TI - An 8-year experience with routine SL mix patch testing supplemented with Compositae mix in Denmark. AB - Routine patch testing with sesquiterpene lactone (SL) mix, supplemented with Compositae mix (CM) and other Compositae extracts and allergens where appropriate, was evaluated over an 8-year period. 190 of 4386 patients tested (4.3%) were Compositae-sensitive, 143 females (mean age 51.5 years) and 47 males (mean age 55 years), and 83% of reactions considered clinically relevant. 22% were suspected of occupational sensitization/dermatitis. 62% had a contact allergy to 2 or more compounds, most often to nickel, fragrance and colophonium. SL mix detected 65%, CM 87% of Compositae-allergic patients, and the overall detection rate with both mixes was 93%. Few irritant reactions and no cases of clear-cut active sensitization were recorded with the mixes, but our results emphasize the importance of differentiating late-appearing reactivation reactions from patch test sensitization. The weakly positive CM reactions could reflect some irritancy, but as they were associated with fragrance and/or colophonium allergy to a higher degree than weakly positive SL mix reactions, they probably represented cross-reactions. In conclusion, the detection rate with SL mix was high enough to support its continued use as a screening mix and it was very well and rather safely supplemented by aimed testing with CM. PMID- 11422266 TI - Allergic contact dermatitis from phenoxyethoxy ethylacrylates in optical fiber coating, and glue in an insulin pump set. PMID- 11422267 TI - Occupational airborne allergic contact dermatitis from succinimidyl carbonates. PMID- 11422268 TI - Contact dermatitis from Cnidoscolus angustidens. PMID- 11422269 TI - Occupational allergic contact dermatitis from N,N-bis(3-aminopropyl)dodecylamine and dimethyldidecylammonium chloride in 2 hospital staff. PMID- 11422270 TI - Oral allergy syndrome from kiwi fruit after a lover's kiss. PMID- 11422271 TI - Connubial allergic contact balanitis due to chlorhexidine. PMID- 11422272 TI - Positive patch test in vancomycin allergy. PMID- 11422273 TI - Nipple dermatitis -- not all what it 'seams'. PMID- 11422274 TI - Occupational nickel dermatitis in fritter making. PMID- 11422275 TI - Sensitization to methyl methacrylate in the plastic catheter of an insulin pump infusion set. PMID- 11422276 TI - Occupational allergic contact urticaria and rhinoconjunctivitis from a detergent protease. PMID- 11422277 TI - Allergic contact dermatitis due to dorzolamide eyedrops. PMID- 11422278 TI - Occupational allergic contact dermatitis from meropenem. PMID- 11422279 TI - Occupational allergic contact dermatitis from tetrahydrofurfuryl acrylate in a medical-device adhesive. PMID- 11422280 TI - Airborne contact dermatitis due to chloroacetamide in wall paint. PMID- 11422281 TI - Allergic contact dermatitis from cycloaliphatic epoxide in jet aviation hydraulic fluid. PMID- 11422282 TI - Allergic contact dermatitis from colophonium in the sawdust of Asturian cider bars. PMID- 11422283 TI - Allergic contact dermatitis from sodium pyrithione in metalworking fluid. PMID- 11422284 TI - Type IV hypersensitivity to oral nystatin. PMID- 11422285 TI - Occupational allergic contact dermatitis in carpenters. PMID- 11422286 TI - Rosemary cheilitis: one to remember. PMID- 11422287 TI - Cell proliferation and protein synthesis as initial factors in determination of axial polarity. AB - The rate of cell proliferation relative to that of protein synthesis appears to have an initial role in establishment of axial polarities in developing animal embryos. An increase in this ratio leads to anterior or dorsal differentiation, while reduction allows posterior or ventral differentiation in a number of organisms. The role that various growth factors play in the regulation of proliferation and protein synthesis is examined. PMID- 11422288 TI - Overexpression of Fyn tyrosine kinase causes abnormal development of primary sensory neurons in Xenopus laevis embryos. AB - The expression and function of the Src family protein tyrosine kinase Fyn in Xenopus laevis embryos have been examined. In situ hybridization analysis demonstrated nervous system-specific expression of Fyn mRNA in tail-bud embryos. However, a class of primary sensory neurons; that is, Rohon-Beard (RB) neurons, which is positive for immunoglobulin superfamily cell adhesion molecules (CAM), neural cell adhesion molecule (N-CAM) and contactin, is devoid of Fyn expression. Injection of Fyn mRNA into one of the blastomeres at the 2-cell stage led to overexpression of Fyn in the injected half of the tail-bud embryos. Immunolabeling of the embryos with anti-HNK-1 antibody revealed that the peripheral axons of RB neurons were partially misguided and bound to each other to form abnormal subcutaneous fascicles. Similar abnormality was induced by injection of the Fyn overexpression vector. The incidence of abnormality appeared dose-dependent, being 68-92% of the injected embryos at 50-400 pg of mRNA. Co injection of the contactin antisense vector depleted contactin mRNA accumulation without affecting Fyn overexpression and reduced the incidence of the abnormal RB cell phenotype. However, the N-CAM antisense was ineffective in reducing this abnormality. These results suggest that Fyn can modify signals regulating axonal guidance or fasciculation in the developing X. laevis nervous system and that contactin may affect this action of Fyn. PMID- 11422289 TI - Left-right asymmetry in Drosophila melanogaster gut development. AB - While left-right (LR) asymmetric morphogenesis is common to various animal species, there have been no systematic studies of the LR asymmetry of body structures of Drosophila melanogaster. In the present paper the LR asymmetric development of the Drosophila gut is described, in which three major parts, the foregut, midgut and hindgut, show almost invariant LR asymmetry. The asymmetry is generated by a twist of each part in particular orientations, resulting in a left handed (sinistral) convolution as a whole. The frequency of spontaneous reversal of LR orientations is very low (< 0.6%) and reversal of each part of the gut occurs independently. The bicoid mutation causes duplication of the posterior half of the gut, essentially keeping the left-handed twist, suggesting that the LR asymmetry may depend on some intrinsic nature of the cells or tissues rather than a graded distribution of morphogens in the egg. The handedness of particular gut parts was randomized or became symmetric in mutants of brachyenteron, huckebein and patched, suggesting that different gene pathways can interfere in determining LR asymmetry of the gut. It is noteworthy that all of these genes are expressed LR symmetrically. PMID- 11422290 TI - Role of a vitelline layer-associated 350 kDa glycoprotein in controlling species specific gamete interaction in the sea urchin. AB - The process of sperm-egg binding is one of the barriers to cross-fertilization between related sea urchin species. A 350 kDa glycoprotein in the egg vitelline layer of Strongylocentrotus purpuratus has been shown to be a sperm-binding protein (SBP). Sulfated O-linked oligosaccharide chains on the 350 kDa glycoprotein, as well as domains of the polypeptide chain, serve as ligands for this binding process. The hypothesis that species-specific sperm-egg binding is attributed to the interaction between the sperm and the 350 kDa glycoprotein was tested using S. purpuratus and S. franciscanus. It was found that both species had a 350 kDa glycoprotein on the egg surface that cross-reacted immunologically using antibodies prepared against a recombinant form of the SBP. Because earlier studies had implicated the carbohydrate chains of the 350 kDa glycoprotein of S purpuratus in sperm binding, differences in carbohydrate chains on the 350 kDa glycoproteins of these species were examined. It was found that among the lectins tested only wheat germ agglutinin and Sambucus nigra agglutinin showed a significant difference in reactivity to the 350 kDa glycoproteins between species. Finally, using a bead-binding assay, it was shown that the isolated 350 kDa glycoproteins exhibited species-specific sperm-binding activity. PMID- 11422291 TI - Two members of the IgLON family are expressed in a restricted region of the developing chick brain and neural crest. AB - The precise expression patterns of two IgLON genes, CEPU-1 and limbic system associated membrane protein (LAMP), were studied during early embryogenesis. It was found that expression of both was localized to restricted regions of the brain and neural crest. In the developing neural tube, CEPU-1 was expressed in the isthmus and a restricted region of the hindbrain, whereas LAMP was expressed in the anterior midbrain. Most neural crest cells expressed LAMP, whereas CEPU-1 expression was limited to crest cells derived from the hindbrain. These results suggest that members of the IgLON family have important roles during embryogenesis, particularly in brain formation and differentiation. PMID- 11422292 TI - Ectoderm exerts the driving force for gastrulation in the sand dollar Scaphechinus mirabilis. AB - How the ectodermal layer relates to the invagination processes was examined in the sand dollar Scaphechinus mirabilis. When the turgor pressure of blastocoele was increased, invagination was completely blocked. In contrast, an increase in turgor pressure did not affect elongation of the gut rudiment in the regular echinoid Hemicentrotus pulcherrimus. Rhodamine-phalloidin staining showed that the distribution of actin filaments was different between two species of embryos. In S. mirabilis gastrulating embryos, abundant actin filaments were seen at the basal cortex of ectoderm in addition to archenteron cells, while the intense signal was restricted to the archenteron in H. pulcherrimus. To investigate whether actin filaments contained in the ectodermal layer exert the force of invagination, a small part of the ectodermal layer was aspirated with a micropipette. If S. mirabilis embryos were aspirated from the onset of gastrulation, invagination did not occur at all, irrespective of the suction site. Even after the archenteron had invaginated to one-half of its full length, further elongation of the archenteron was severely blocked by suction of the lateral ectoderm. In contrast, suction of the ectodermal layer did not affect the elongation processes in H. pulcherrimus. These results strongly suggest that the ectodermal layer, especially in the vegetal half, exerts the driving force of invagination in S. mirabilis. PMID- 11422293 TI - Spatial expression patterns of genes involved in cyclic AMP responses in Dictyostelium discoideum development. AB - The spatial expression patterns of genes involved in cyclic adenosine monophosphate (cAMP) responses during morphogenesis in Dictyostelium discoideum were analyzed by in situ hybridization. Genes encoding adenylyl cyclase A (ACA), cAMP receptor 1, G-protein alpha2 and beta subunits, cytosolic activator of ACA (CRAC and Aimless), catalytic subunit of protein kinase A (PKA-C) and cAMP phosphodiesterases (PDE and REG-A) were preferentially expressed in the anterior prestalk (tip) region of slugs, which acts as an organizing center. MAP kinase ERK2 (extracellular signal-regulated kinase-2) mRNA, however, was enriched in the posterior prespore region. At the culmination stage, the expression of ACA, CRAC and PKA-C mRNA increased in prespore cells in contrast with the previous stage. However, no alteration in the site of expression was observed for the other mRNA analyzed. Based on these findings, two and four classes of expression patterns were catalogued for these genes during the slug and culmination stages, respectively. Promoter analyses of genes in particular classes should enhance understanding of the regulation of dynamic and coordinated gene expression during morphogenesis. PMID- 11422294 TI - A novel putative transmembrane protein, IZP6, is expressed in neural cells during embryogenesis. AB - Gene trapping in mouse embryonic stem cells is an efficient method for identifying new genes and examining their functions. This method has been used in an effort to identify some novel genes involved in mouse development. In the present paper, one such gene named IZP6 is reported. Expression of the IZP6 gene, as monitored by beta-galactosidase expression in heterozygous mice, was detected in a developmentally regulated fashion: the expression pattern has two phases during the embryogenesis. In the first phase, from embryonic day 11.5 (E11.5) until E14.5, the reporter gene is mainly expressed in the forebrain. In the second phase, from E15.5 until birth, expression in the forebrain becomes weaker but is still observed in the olfactory bulb and the skin around the eyes, nose, limbs and tail. Thus, IZP6 gene expression changes from the central nervous system (the first phase) to the peripheral tissues (the second phase) during development. The IZP6 gene encodes a protein of 228 amino acids. Analysis of the secondary structure of the IZP6 protein revealed four hydrophobic regions, indicating that the IZP6 protein is a four transmembrane region protein. These results suggest that IZP6 is a transmembrane protein related to neurogenesis in the mouse. PMID- 11422295 TI - Nucleus: cell volume ratio directs the timing of the increase in blastomere adhesiveness in starfish embryos. AB - Blastomeres of starfish embryos begin to increase in adhesiveness after the eighth cleavage and form a monolayered hollow blastula. To investigate factors that affect the timing of the adhesiveness increase, we changed the volume of the cytoplasm or the ploidy of embryos and examined the morphologic changes in the descendent blastomeres during early cleavage stages. In parthenogenetic embryos, in which the ploidy is doubled, the timing of the increase in adhesiveness was accelerated by one cell cycle. In contrast, the timing was delayed by approximately one cell cycle in a large-sized embryo formed by the fusion of an egg and a non-nucleate egg fragment. These two sets of observations are in accord with the expectation from the classical concept that the DNA: cytoplasmic ratio may direct the timing of events in early development. However, observations of small-sized embryos with a reduced amount of cytoplasm were contradictory to the expectation based on the DNA: cytoplasmic ratio; the timing of the increase in adhesiveness in half-sized embryos was almost the same as in control embryos and the timing was delayed by only one cell cycle in quarter-sized embryos. Measurement of the diameters of nuclei showed that the size of nuclei was variable, depending on the stage of development, the volume of cytoplasm and ploidy. We calculated a volume ratio of nucleus to cytoplasm (N: C volume ratio) for tetraploid, large-, half- and quarter-sized embryos. We found that the embryonic cells begin to adhere always when their N: C volume ratio reaches 0.06. A plausible model for the cellular timing mechanism of cell contact is proposed. PMID- 11422296 TI - A hatching enzyme substrate in the Xenopus laevis egg envelope is a high molecular weight ZPA homolog. AB - The Xenopus laevis egg envelope is composed of six or more glycoproteins, three of which have been cloned and identified as the mammalian homologs ZPA (ZP2), ZPB (ZP1) and ZPC (ZP3). The remaining glycoproteins are a triplet of high molecular weight components that are selectively hydrolyzed by the hatching enzyme. We have isolated one of these proteins and cloned its cDNA. The mRNA for the protein was found to be expressed only in early stage oocytes, as are other envelope components. From the deduced amino acid sequence, it was indicated to be a secreted glycoprotein with a characteristic ZP domain in the C-terminal half of the molecule. The N-terminal half was unrelated to any known glycoprotein. Comparative sequence analysis of the ZP domain indicated that it was derived from an ancestor of ZPA and ZPB, with the greatest identity to ZPA. This envelope component has been designated ZPAX. PMID- 11422297 TI - Effects of microgravity on the larval development, metamorphosis and reproduction of the urodele amphibian Pleurodeles waltl. AB - The FERTILE experiment was twice performed onboard the Mir space station during the Cassiopee and Pegase French space missions. The goal was to analyze the effects of microgravity on fertilization and embryonic development, and then on further development on the ground in the amphibian Pleurodeles waltl. The present paper reports development that occurred in the laboratory after landing. Recovered on the ground at the hatching stage, young larvae reared at room temperature underwent metamorphosis and became adults without obvious abnormalities. Of particular interest was the rearing temperature that induced a delayed metamorphosis for animals from the Cassiopee space mission, but not for animals from the Pegase mission. The rate of development and the morphology were analogous in these animals and in ground controls reared in a similar annual period. Analysis of offspring was performed using these animals. Males born in space were first mated with control ground-born females and then with females born in space. The mating gave progeny that developed normally. Depending on the methods used and on the limits of the analyses, the results clearly demonstrated that animals born in space were able to live and reproduce after return to the ground. PMID- 11422298 TI - Mechanical sutures in esophageal replacement: fashion or resource? PMID- 11422299 TI - Jejunum interposition after cervical esophageal resection. PMID- 11422300 TI - Adenocarcinoma of the rat esophagus in the presence of a proton pump inhibitor: a pilot study. AB - This study examines the effects of a proton pump inhibitor on a rat model of duodenogastric reflux. Duodenoesophageal reflux was induced in 60 rats by performing a duodenesophagostomy. The study group received daily intraperitoneal injections of a proton pump inhibitor for 6 months and the control group received an equivalent injection of saline. Rats were examined at death for macroscopic tumor, dysplasia, adenocystic changes, papillomatosis, and adenocarcinoma. Five out of 19 rats in the study group and three out of 20 rats in the control group developed dysplastic/adenocarcinomatous changes. Ten of the rats in the study group died before the end of the study, as opposed to one in the control group (this is not statistically significant). There was no difference in the number of cancers that developed in the two groups. However, there was an insignificant trend to earlier appearance of detectable disease in the study group. PMID- 11422301 TI - Acidic fibroblast growth factor is expressed sequentially in the progression from Barrett's esophagus to esophageal adenocarcinoma. AB - Acidic fibroblast growth factor 1 (FGF-1) is sequentially accumulated in Barrett's esophagus and its expression in glandular dysplasias is independent of esophageal adenocarcinoma. This suggests that FGF-1 immunohistochemistry could be used as an adjunct to the routine histopathologic diagnosis of dysplasia in Barrett's esophagus. The data also underscore the important role of fibroblast growth factors in tumorigenesis. PMID- 11422302 TI - Localization of small esophageal cancers for radiation planning using endoscopic contrast injection: report on a series of eight cases. AB - Recently, Barrett's esophagus and early adenocarcinomas have been detected increasingly frequently in routine follow-up of patients with gastroesophageal reflux. Although surgery is the treatment of choice, some patients are medically unfit for esophagectomy and, in this case, the only alternative curative therapy is radical chemoradiation therapy. In addition, some patients who present with symptoms have small tumors that cannot be localized accurately using routine imaging techniques. This report describes a series of eight patients with small esophageal cancers in whom the tumors were successfully localized following endoscopic injection of contrast, and treated with chemoradiation therapy. The treatment was successful in seven patients. This method of tumor localization demonstrated that conventional techniques are mostly unreliable when applied to very early cancers. PMID- 11422303 TI - Ratio of invaded to removed lymph nodes as a prognostic factor in adenocarcinoma of the distal esophagus and esophagogastric junction. AB - This study examined the influence of nodal harvest and the proportion of positive nodes on survival in 59 patients with adenocarcinoma of the distal esophagus and esophagogastric junction undergoing esophagectomy with curative intent. A total of 754 lymph nodes were harvested (median 13, range 0-28). Two hundred and twenty eight positive nodes were found on histology (median 4, range 1-23) in 43 (79%) patients with a higher incidence from T3/T4 than T1/T2 lesions (P < 0.003). Overall 1- and 3-year survival rates were 73% and 47% respectively. Node positivity increased with increased total nodal harvest, but was not influenced by the site of tumors or surgical approaches. There was no survival benefit for patients with <20% over >20% nodal positivity (P=0.31). Only negative lateral resection margin emerged as a significant factor in both univariate (P < 0.01) and multivariate analysis (P < 0.05). We conclude that the degree of nodal positivity in adenocarcinoma is less important than resection margin status as a prognostic factor. PMID- 11422304 TI - Ketoconazole and pulmonary failure after esophagectomy: a prospective clinical trial. AB - Thromboxane is a key mediator in pulmonary injury after esophageal resection. In this prospective trial we studied the clinical course and development of pulmonary alterations in patients undergoing esophagectomy and prophylactic treatment with a thromboxane synthase inhibitor. Thirty-eight consecutive patients undergoing esophageal resection were treated pre- and perioperatively with 3 x 200 mg ketoconazole. The clinical course was studied and pulmonary function was assessed according to the Murray score. A historical group of 118 patients undergoing esophagectomy for benign and malignant esophageal diseases served as controls. Patients in both groups were similar in terms of age, sex, and preoperative pulmonary function, as well as in the anesthetic and surgical procedures performed. However, in the ketoconazole group, more patients were at risk of pulmonary failure by receiving neoadjuvant radiochemotherapy (22/38) or undergoing thoracotomies (33/38) than control subjects (14/118 and 80/118, P < 0.05). Two out of 38 ketoconazole-treated patients developed acute lung injury after esophagectomy, as did 20/118 control patients (P < 0.05). This prospective non-randomized clinical study (in patients subjected to esophagectomy) provides further evidence that prophylactic thromboxane synthase inhibition by ketoconazole reduces the incidence of acute lung injury in patients at risk. PMID- 11422305 TI - Gastroesophageal reflux disease: clinical, endoscopic, and intraluminal esophageal pH monitoring evaluation. AB - One hundred and twenty-two patients with gastroesophageal reflux disease were studied (90 with and 32 without esophagitis) with the objective of analyzing possible differences between those with and without esophagitis. Evaluation consisted of clinical interview, endoscopy of the high digestive tract, esophageal manometry, and pH monitoring. There was no significant difference between the groups in age, sex, or symptoms. The incidence of hiatal hernia was greater in the group with esophagitis. Although the frequency of motor changes was similar, the type of anomaly was different. The reflux pattern was very similar in both groups. Therefore, the concept of reflux disease, esophagitis, and pathological reflux still needs a broader definition for greater diagnostic precision and for comparing the results of different studies on the subject. Normal reflux (confirmed using pH esophageal monitoring) in 12.2% of patients with esophagitis suggests that other factors are implicated in the etiology of the disease besides those measured using this examination. PMID- 11422306 TI - Obesity and its effect on outcome of laparoscopic Nissen fundoplication. AB - Obesity has long been suspected as predisposing to gastroesophageal reflux disease, and it has also been claimed that it is an important cause of poor outcome following laparoscopic anti-reflux surgery. This study was performed to determine the validity of this proposition. The outcome of 194 patients from an overall experience of 971 laparoscopic anti-reflux procedures was determined in this study. Patients were included if they had undergone a laparoscopic Nissen fundoplication, had completed a minimum 12 months follow-up using a structured questionnaire, and had data available for the calculation of their preoperative body mass index (BMI). Patients were divided into three groups based on BMI: normal weight (BMI < 25), overweight (BMI 25-29.9), and obese (BMI >30). The association between BMI and outcome data from their most recent follow-up was analyzed. There was no correlation between increasing BMI and a poorer overall outcome. There was a slight trend toward less satisfaction with the surgical outcome in patients of normal weight. Preoperative obesity is not associated with a poorer outcome following laparoscopic Nissen fundoplication. PMID- 11422307 TI - Vector volume analysis of the lower esophageal sphincter in achalasia and the effect of balloon dilation. AB - Assessment of the effect of balloon dilation of the lower esophageal sphincter in achalasia currently relies on symptom scores, radiologic studies, or manometry. We have used vector volume analysis which constructs a three-dimensional (3D) pressure profile to examine the effects of balloon dilation on the sphincter. Studies were performed in 26 normal subjects and in 11 patients with achalasia in whom vector volume was determined before and/or after balloon dilation. Vector volume analysis was performed using a multilumen manometry catheter with eight side-reading ports. The catheter was withdrawn through the sphincter using a continuous pull-through technique. A 3D pressure profile was constructed. Vector volume of the lower esophageal sphincter shows a marked radial and linear asymmetry in normal subjects. 3D pressure profiles showed a symmetrical increase in lower esophageal sphincter pressure in untreated achalasia. Mean vector volumes pre and postdilation fell from 254 to 88 cm3 respectively. This was associated with a fall in the mean radial symmetry index from 0.83 to 0.76. Improvement in dysphagia occurred in those patients in whom pressure fell to 15 mmHg or below in at least three adjacent segments. This technique may be a useful tool in the assessment of patients with achalasia and postdilation results may help to predict the long-term outcome of treatment. PMID- 11422308 TI - Abnormal esophagocardiac reflex in patients with non-cardiac chest pain. AB - As the mechanoreceptor stimulation of the esophagus activates an esophagocardiac inhibitory reflex, with possible cardiac hypokinetic arrhythmias, we investigated whether patients with non-cardiac chest pain have this reflex, which could represent a source of risk in predisposed individuals during the intraesophageal balloon distension test. Electrocardiogram readings were recorded in nine patients with non-cardiac chest pain (group A), the esophageal origin of which was diagnosed with cardiac and esophageal examinations, in 10 patients with hyperkinetic esophageal motor disorders without chest pain (group B), and in eight normal subjects used as controls (group C), after swallowing solid boluses and during intraesophageal balloon inflation at 100 mmHg for 10 s. The percent variation of the R-R interval from its mean basal value to its highest value observed after stimulation was calculated. Solid swallows induced an increase in heart rate followed by a decrease that was significantly higher in group B than group C, while group A was not significantly different from group C. Balloon inflation induced a significant decrease in heart rate in all groups, but in group A the degree of decrease was significantly lower than in groups B and C. In conclusion, esophageal wall distension, either as a result of solid bolus or balloon inflation, elicits an inhibitory esophagocardiac reflex that is higher than normal in patients with hyperkinetic esophageal motor disorders without pain and lower than normal in patients with non-cardiac chest pain of esophageal origin, who, consequently, have nothing to fear from this procedure. PMID- 11422309 TI - Epiphrenic diverticula: minimal invasive approach and repair in five patients. AB - Epiphrenic esophageal diverticula are rare and often asymptomatic. If surgery is mandatory, a thoracotomy is used to resect the diverticulum. The results of a minimal invasive approach and repair in five patients are presented. These patients, who all presented with an epiphrenic diverticulum, were evaluated using barium swallow study, esophagoscopy, and manometry. The diverticula were approached by thoracoscopy in all patients and a description of the surgical technique is given. The diverticula were resected using a right-sided approach in four patients. One patient with a diverticulum in the distal esophagus required conversion to laparoscopy. A myotomy was performed in two patients because of high pressures in the lower esophageal sphincter. The postoperative course was uncomplicated in four patients. One patient with Ehlers-Danlos disease had a complicated course owing to leakage, resulting in two re-operations by means of thoracotomy. There was no mortality. The minimal invasive approach of epiphrenic diverticula is feasible. The long-term results are awaited. PMID- 11422310 TI - Manometric study in Kearns-Sayre syndrome. AB - Although swallowing difficulties have been described in patients with Kearns Sayre syndrome (KSS), the spectrum of manometric characteristics of dysphagia is not yet well known. Moreover, it is conceivable that a combination of various degrees of swallowing difficulties with different patterns in manometric studies exist, each playing a major role in the prognosis, natural history, and quality of life of KSS patients. An 18-year-old girl diagnosed at the age of 5 years with KSS (muscle biopsy) was admitted to our department with an upper respiratory tract infection and dysphagia. Clinical examination revealed growth retardation, external ophthalmoplegia, pigmentary retinopathy, impaired hearing, and ataxia. An electrocardiogram revealed cardiac conduction defects (long Q-T), and brain magnetic resonance imaging showed abnormalities in the cerebellar hemispheres. A manometric and motility study for dysphagia was conducted and the pharynx and upper esophageal sphincter (UES) resting pressures were similar to control group values, but the swallowing peak contraction pressure of the pharynx and the closing pressure of the UES were very low and could not promote effective peristaltic waves. Relaxation and coordination of the UES were not affected although pharyngeal and upper esophagus peristaltic waves proved to be very low and, consequently, were practically ineffective. The patient was started on treatment comprising a diet rich in potassium, magnesium, and calcium, and oral administration of vitamin D and co-enzyme Q10 100 mg daily; she was discharged 6 days later with apparent clinical improvement. PMID- 11422311 TI - Chylous ascites complicating esophagectomy. PMID- 11422312 TI - Adenocarcinoma of the esophagus with choroidal metastasis. AB - In this report, a case is presented of an adenocarcinoma in a Barrett's esophagus metastatic to the choroid. A 54-year-old woman presented with a rapidly progressive decrease of vision in the right eye 8 months after intentionally curative esophagectomy for an adenocarcinoma. Fundoscopy, ultrasonography, and magnetic resonance imaging findings were suggestive of a metastasis. The patient received palliative external beam irradiation to the right eye for visual restoration, but she died before any beneficial effect was achieved. PMID- 11422313 TI - Laparoscopic management of pseudoachalasia, esophageal diverticulum, and benign esophageal stromal tumor. AB - A case report is presented of a 59-year-old woman who was suspected of having a paraesophageal hernia, but at operation was found to have an epiphrenic diverticulum of the esophagus, a benign stromal tumor of the esophagus, and pseudoachalasia. The stromal tumor was resected laparoscopically together with a laparoscopic Heller's myotomy and partial posterior fundoplication. PMID- 11422314 TI - Perforation of a gastric tube peptic ulcer into the thoracic aorta. AB - We present a case of a 52-year-old male patient who died from massive hematemesis as a result of perforation of a benign peptic ulcer into the descending thoracic aorta, 1 year after esophagectomy for esophageal cancer and gastric tube interposition. We also review the literature for mechanisms of ulceration in intrathoracic gastric grafts and for complications of such ulcers. PMID- 11422315 TI - Esophageal replacement by an isoperistaltic stomach tube. PMID- 11422316 TI - Thermophilic methanogens in rice field soil. AB - The soil temperature in flooded Italian rice fields is generally lower than 30 degrees C. However, two temperature optima at approximately 41 degrees C and 50 degrees C were found when soil slurries were anoxically incubated at a temperature range of 10-80 degrees C. The second temperature optimum indicates the presence of thermophilic methanogens in the rice field soil. Experiments with 14C-labelled bicarbonate showed that the thermophilic CH4 was exclusively produced from H2/CO2. Terminal restriction fragment length polymorphism (T-RFLP) of archaeal SSU rRNA gene fragments revealed a dramatic change in the archaeal community structure at temperatures > 37 degrees C, with the euryarchaeotal rice cluster I becoming the dominant group (about 80%). A clone library of archaeal SSU rRNA gene fragments generated at 49 degrees C was also dominated (10 out of 11 clones) by rice cluster I. Our results demonstrate that Italian rice field soil contains thermophilic methanogenic activity that was most probably a result of members of the as yet uncultivated euryarchaeotal rice cluster I. PMID- 11422317 TI - Linking the composition of bacterioplankton to rapid turnover of dissolved dimethylsulphoniopropionate in an algal bloom in the North Sea. AB - The algal osmolyte, dimethylsulphoniopropionate (DMSP), is abundant in the surface oceans and is the major precursor of dimethyl sulphide (DMS), a gas involved in global climate regulation. Here, we report results from an in situ Lagrangian study that suggests a link between the microbially driven fluxes of dissolved DMSP (DMSPd) and specific members of the bacterioplankton community in a North Sea coccolithophore bloom. The bacterial population in the bloom was dominated by a single species related to the genus Roseobacter, which accounted for 24% of the bacterioplankton numbers and up to 50% of the biomass. The abundance of the Roseobacter cells showed significant paired correlation with DMSPd consumption and bacterioplankton production, whereas abundances of other bacteria did not. Consumed DMSPd (28 nM day(-1)) contributed 95% of the sulphur and up to 15% of the carbon demand of the total bacterial populations, suggesting the importance of DMSP as a substrate for the Roseobacter-dominated bacterioplankton. In dominating DMSPd flux, the Roseobacter species may exert a major control on DMS production. DMSPd turnover rate was 10 times that of DMS (2.7 nM day(-1)), indicating that DMSPd was probably the major source of DMS, but that most of the DMSPd was metabolized without DMS production. Our study suggests that single species of bacterioplankton may at times be important in metabolizing DMSP and regulating the generation of DMS in the sea. PMID- 11422318 TI - Transcript and activity levels of different Pleurotus ostreatus peroxidases are differentially affected by Mn2+. AB - The white-rot fungus Pleurotus ostreatus produces both manganese-dependent peroxidase (MnP) and versatile peroxidase (VP) in non-manganese-amended peptone medium (PM). We studied the effect of Mn2+ supplementation on MnPs and VPs in P. ostreatus by analysing the enzymatic and transcript abundance profiles of the peroxidases, as well as the lignin mineralization rate. The fungus was grown in PM under solid-state conditions using perlite as an inert solid support. Mn2+ amendment resulted in a 1.7-fold increase in [14C]-lignin mineralization relative to unamended medium. Anion-exchange chromatography was used to resolve the fungal peroxidase's enzymatic activity profile. Five peaks (P1-P5) of VP and one peak (P6) of MnP activity were detected in unamended medium. In Mn2+-amended medium, a reduction in the activity of the VPs was observed. On the other hand, a sharp increase in the MnP activity level of peak P6 was detected. The P6 isoenzyme was purified and showed manganese-dependent peroxidation of phenolic substrates. Internal sequence analysis of the purified enzyme revealed 100% identity with the deduced amino acid sequence of P. ostreatus MnP3 (GenBank AB016519). The effect of Mn2+ on the relative abundance of gene transcripts of three VPs and one MnP from P. ostreatus was monitored using reverse transcription-polymerase chain reaction (RT-PCR) with oligonucleotide primer sets synthesized on the basis of non-conserved sequences of the different peroxidases. The reduction in VP gene transcript abundance and the increase in mnp3 transcript level were collinear with the changes observed in the enzyme activity profiles. These results indicate that the activity of peroxidases is regulated at the transcriptional level. We suggest that the expression of MnP and VP may be differentially regulated by the presence of Mn2+. PMID- 11422319 TI - Phylogenetic analysis of ribosomal RNA operons from uncultivated coastal marine bacterioplankton. AB - Analyses of small subunit ribosomal RNA genes (SSU rDNAs) have significantly influenced our understanding of the composition of aquatic microbial assemblages. Unfortunately, SSU rDNA sequences often do not have sufficient resolving power to differentiate closely related species. To address this general problem for uncultivated bacterioplankton taxa, we analysed and compared sequences of polymerase chain reaction (PCR)-generated and bacterial artificial chromosome (BAC)-derived clones that contained most of the SSU rDNAs, the internal transcribed spacer (ITS) and the large subunit ribosomal RNA gene (LSU rDNA). The phylogenetic representation in the rRNA operon PCR library was similar to that reported previously in coastal bacterioplankton SSU rDNA libraries. We observed good concordance between the phylogenetic relationships among coastal bacterioplankton inferred from SSU or LSU rDNA sequences. ITS sequences confirmed the close intragroup relationships among members of the SAR11, SAR116 and SAR86 clades that were predicted by SSU and LSU rDNA sequence analyses. We also found strong support for homologous recombination between the ITS regions of operons from the SAR11 clade. PMID- 11422320 TI - Lipopolysaccharide O-chain microheterogeneity of Salmonella serotypes Enteritidis and Typhimurium. AB - Variability in the lipopolysaccharide (LPS) of the two most prevalent Salmonella serotypes causing food-borne salmonellosis was assessed using gas chromatography analysis of neutral sugars from 43 Salmonella enterica serovar Enteritidis (S. Enteritidis) and 20 Salmonella enterica serovar Typhimurium (S. Typhimurium) isolates. Four substantially different types of O-chain chemotypes were detected using cluster analysis of sugar compositions; these were low-molecular-mass (LMM) LPS, glucosylated LMM LPS, high-molecular-mass (HMM) LPS and glucosylated HMM LPS. Nineteen out of 20 S. Typhimurium isolates yielded glucosylated LMM. In contrast, S. Enteritidis produced a more diverse structure, which varied according to the source and history of the isolate: 45.5% of egg isolates yielded glucosylated HMM LPS; 100% of stored strains lacked glucosylation but retained chain length in some cases; and 83.3% of fresh isolates from the naturally infected house mouse Mus musculus produced glucosylated LMM LPS. A chain length determinant (wzz) mutant of S. Enteritidis produced a structure similar to that of S. Typhimurium and was used to define what constituted significant differences in structure using cluster analysis. Fine mapping of the S. Enteritidis chromosome by means of a two-restriction enzyme-ribotyping technique suggested that mouse isolates producing glucosylated LMM LPS were closely related to orally invasive strains obtained from eggs, and that stored strains were accumulating genetic changes that correlated with suppression of LPS O-chain glucosylation. These results suggest that the determination of LPS chemotype is a useful tool for epidemiological monitoring of S. Enteritidis, which displays an unusual degree of diversity in its LPS O-chain. PMID- 11422321 TI - Nitrogenase activity in cyanobacteria measured by the acetylene reduction assay: a comparison between batch incubation and on-line monitoring. AB - A new on-line method for measuring acetylene reduction is described. It consists of a gas-flow cell connected to an electronic gas-mixing system and an automatic sample loop in the gas chromatograph. Alternatively, ethylene can be determined by using laser-based trace gas detection. The laser-based trace gas detection technique achieves a detection limit that is three orders of magnitude better than gas chromatography. We have applied the on-line method to the measurement of nitrogen fixation in a culture of the heterocystous cyanobacterium Nodularia spumigena and compared it with conventional batch-type incubations. Incubation of N. spumigena in the gas-flow cell resulted in very short response times with a steady-state flux of ethylene obtained within 2 min. Nitrogenase was shown to respond immediately to changes in light and oxygen. Monitoring of nitrogenase activity could be continued for several hours without having a negative impact on nitrogen fixation rates in N. spumigena. This was not the case in batch incubations, in which changes in nitrogenase activities were recorded during incubations, probably as a result of varying oxygen concentrations. It was therefore concluded that the on-line method is superior to batch incubations when rates of nitrogenase activity are to be measured. The method is suitable for natural samples (water or sediment). PMID- 11422322 TI - Soil DNA and the microbial metagenome. An annotated selection of World Wide Web sites relevant to the topics in Environmental Microbiology Web alert. PMID- 11422323 TI - Usefulness of the head-upright tilt test for distinguishing syncope and epilepsy in children. AB - PURPOSE: Episodic loss of consciousness in children, whether or not associated with hypertonia or short-duration clonic movements, presents a diagnostic challenge to the pediatrician and child neurologist. We provide some evidence of the usefulness of the head-upright tilt test for investigating the causes of transient loss of consciousness in children, and for distinguishing between syncope, convulsive syncope, and epilepsy. METHODS: We studied nine children previously diagnosed as epileptic on the basis of compatible clinical events and epileptiform findings in routine EEGs who were treated over the long term with antiepileptic drugs, but whose clinical records suggested syncope or convulsive syncope rather than epilepsy on reevaluation. All subjects underwent head-upright tilt testing. RESULTS: The tilt-test result was positive in all nine cases, with the patients reporting the same symptoms as in the previously considered epileptic attacks. CONCLUSIONS: Inadequate histories and misuse/overinterpretation of EEG results often lead to misdiagnosis of epilepsy in children. The head-upright tilt test is a useful and reliable diagnostic technique, allowing syncopal events to be induced and evaluated under controlled conditions. In a subset of patients, it may help to distinguish epilepsy from simple or convulsive syncope. PMID- 11422324 TI - Incidence of status epilepticus in adults in Germany: a prospective, population based study. AB - PURPOSE: To determine the incidence and case-fatality rate of status epilepticus (SE) in adults in Hessen, Germany, we performed a prospective, population-based study from July 1997 through June 1999. METHODS: All adult patients residing within the zip-code area 35 (area-35) with SE were included. Area-35 had 743.285 adult inhabitants, including 123.353 adult inhabitants of the primary service area of the University Hospital Marburg (PS-area). Patients were reported by 16 hospitals in the area and were prospectively identified and carefully reviewed within 5 days by one of the authors. Based on the crude annual incidence of SE and a rate of underascertainment of 10% determined for the PS-area, the corrected, age-adjusted incidence of SE in area 35, more representative of the population of Germany, was calculated. RESULTS: The crude annual incidence in the PS-area was 15.8/100,000 [95% confidence interval (CI), 11.2-21.6]. The calculated, corrected, age-adjusted incidence of SE in area 35 was 17.1/100,000. It was higher for men compared with women (26.1 vs. 13.7) and for those aged 60 years and older (54.5 vs. 4.2/100,000, p < 0.0001). The etiology was mainly remote symptomatic due to cerebrovascular disease. Epilepsy was previously diagnosed in only 50% of the patients. The case-fatality rate was 9.3%. CONCLUSIONS: Based on our data, at least 14,000 patients would be affected by SE in Germany, associated with approximately 1,300 deaths annually. The incidence of SE in Germany is similar to that found in the white United States population. Furthermore, this study confirms the higher incidence of SE in male patients and in the elderly population. This may be due to a higher incidence of cerebrovascular disease in these subpopulations. PMID- 11422325 TI - Acute psychosis during intracranial EEG monitoring: close relationship between psychotic symptoms and discharges in amygdala. AB - PURPOSE: This report examined the underlying mechanism of psychosis associated with epilepsy. METHODS: An adult patient with epilepsy manifesting acute psychosis during long-term EEG monitoring is presented, together with a literature review on this subject. RESULTS: A 25-year-old woman with intractable temporal lobe epilepsy developed acute psychosis while she underwent long-term intracranial EEG monitoring. After a clustering of seizures, she manifested psychotic symptoms including hallucination, stupor, and repeated fear. The transition of psychotic symptoms corresponded to the changes in frequency and morphology of seizure discharges restricted to the left amygdala. Improvement of psychosis coincided with disappearance of seizure discharges. CONCLUSIONS: This case confirmed a close relationship between psychotic symptoms and seizure discharges in the left amygdala. It is suggested that paroxysmal bombardment of the medial temporal lobe structure may be a pathogenetic factor of acute psychosis associated with epilepsy. PMID- 11422326 TI - Distinguishing subtypes of temporal lobe epilepsy with background hippocampal activity. AB - PURPOSE: Two subtypes of temporal lobe epilepsy (TLE) can be defined through clinical observations and analysis of hippocampal tissue resected during surgical procedures for intractable TLE: (a) mesial temporal sclerosis (MTS), which is characterized by extensive changes to the hippocampus and good surgical outcome; and (b) paradoxical temporal lobe epilepsy (PTLE), which is characterized by minimal cell loss and comparatively poorer surgical outcome. Patients in both subtypes have seizures that appear to begin in the medial temporal lobe, but documented differences in substrate and outcome between these subtypes has defined a need to distinguish MTS and PTLE patients before surgery. This report describes a retrospective study to investigate the feasibility of doing so during intracranial monitoring. METHODS: Background EEG epochs, 5 min in duration, were recorded from the anterior hippocampus in 14 (10 MTS and four PTLE) patients with consistent localization of seizure onset to medial temporal structures. The power spectral density (PSD) of the EEG epochs was calculated by a Fourier spectral estimator, and the total signal power and power of the delta, theta, alpha, beta, and gamma frequency bands were submitted to group-to-group comparison. RESULTS: Spectral peaks were observed in the delta band in all PSD estimates and in the theta band in nine of 14 (seven MTS, two PTLE) estimates. The MTS and PTLE subtypes could be distinguished by the total signal power and delta band power. These power measurements were greater in the PTLE subtype. CONCLUSIONS: Both delta and theta spectral components are present in hippocampal background EEGs recorded from patients with TLE. The results indicate that group differences exist in spectral measures of background hippocampal signals recorded from MTS and PTLE subtypes. This suggests both that substrate differences in cellular composition and connectivity are reflected in hippocampal background EEGs and that spectral measurements of these signals may hold promise for tests to identify the group membership of individual patients. PMID- 11422327 TI - Is amygdalohippocampectomy really selective in medial temporal lobe epilepsy? A study using positron emission tomography with (18)fluorodeoxyglucose. AB - PURPOSE: Selective amygdalohippocampectomy (SAH) is a surgical technique effective for the treatment of medial temporal lobe epilepsy, which selectively removes the epileptogenic hippocampus and amygdala but spares the temporal neocortex. However, the benefit of SAH in terms of functional outcome is debated. In this study, we aimed to assess the metabolic consequences of SAH. METHODS: Volumetric magnetic resonance imaging (MRI) and fluorodeoxyglucose positron emission tomography (PET) studies were performed in nine patients with medial temporal lobe epilepsy associated with hippocampal sclerosis before and after SAH. Regions of interest were delineated on MRIs and then replaced on PET images using an automatic 3D image registration. We calculated absolute metabolic rates of glucose and normalized metabolic values in each region of interest. RESULTS: The comparison between preoperative and postoperative metabolic values showed a statistically significant worsening of the hypometabolism on the ipsilateral temporal pole on the superior and the hippocampal levels (p < 0.05 and 0.0045, respectively). A postoperative increase of the metabolic activity also was noted in the contralateral anterior hippocampus (p < 0.05) and the orbitofrontal cortex bilaterally (p < 0.002 and 0.001, respectively) CONCLUSIONS: SAH functional benefit is controversial. SAH worsened significantly the hypometabolism of a temporal structure that was not surgically removed (i.e., the temporal pole), and it improved postoperatively the metabolic activity in the contralateral hippocampus and the orbitofrontal cortex. Whether this postoperative improvement is linked to the selectivity of the surgical procedure must be further clarified. PMID- 11422328 TI - The regulation of serum sodium after replacing carbamazepine with oxcarbazepine. AB - PURPOSE: To evaluate changes in serum electrolyte balance and underlying regulatory mechanisms in 10 male patients with epilepsy 2 and 6 months after replacing long-term carbamazepine (CBZ) monotherapy with oxcarbazepine (OCBZ) monotherapy. Arginine vasopressin (AVP) is thought to be most important underlying mechanism of CBZ-related hyponatremia via direct or kidney tubular mechanisms. Furthermore, AVP is as well hormonally regulated by the renin angiotensin-aldosterone system and atrial natriuretic peptide (ANP). METHODS: The medication of the patients was changed from CBZ to OCBZ. Serum electrolytes, creatinine, albumin, aldosterone, and the N-terminal fragment of ANP (NT-proANP) concentrations were measured before and 2 and 6 months after the change in the medication. RESULTS: The mean serum sodium level diminished after the medication was changed. Serum sodium levels decreased below the reference range in two (20%) patients during OCBZ medication. Serum sodium levels decreased altogether in four patients, and remained unaltered in six patients. Serum aldosterone levels increased in the six patients whose serum sodium concentrations did not decrease, but no increase was found in the patients with decreased sodium levels during OCBZ medication. Serum NT-proANP levels decreased in all patients. CONCLUSIONS: Serum sodium levels decrease after replacing CBZ with OCBZ. The low serum NT proANP concentrations appear to reflect the decreased serum sodium levels, but a compensatory aldosterone response may prevent the development of hyponatremia in some patients during OCBZ medication. PMID- 11422329 TI - Acetazolamide in women with catamenial epilepsy. AB - PURPOSE: Catamenial epilepsy is a condition characterized by an increase in seizures during particular phases of the menstrual cycle. The incidence of catamenial epilepsy varies widely, partly because of a lack of a universally adopted definition. Specific treatment options for these patients are limited. The use of acetazolamide (AZ) has been based largely on anecdotal reports demonstrating efficacy in small or poorly characterized populations. The purpose of this study was to analyze retrospectively the efficacy, safety profile, and tolerability of AZ in women with catamenial epilepsy. METHODS: Women with catamenial epilepsy identified from 1990 through 1999 were invited to participate in a retrospective telephone questionnaire addressing the relationship of seizures and the menstrual cycle and the use, efficacy, and adverse effects of AZ. Seizure outcome was classified as: seizure free (SF), significantly improved (SI), or not significantly improved (NSI). Responses to AZ were compared in women with different types of epilepsy and comparing continuous versus intermittent dosing using Fisher's exact tests. RESULTS: Twenty women were identified who had received or were currently taking AZ. The drug was given continuously in 55% and intermittently in 45% of patients. A > or =50% decrease in the seizure frequency was reported by 40% of subjects. Response rates were similar in generalized and focal epilepsy and in temporal and extratemporal epilepsy. There was no significant difference in effectiveness comparing continuous with intermittent dosing. A loss of efficacy over 6-24 months was reported by 15% of women. CONCLUSIONS: Despite our small sample and retrospective design, AZ appears to demonstrate efficacy for catamenial epilepsy. PMID- 11422330 TI - Repeated neural tube defects and valproate monotherapy suggest a pharmacogenetic abnormality. AB - Valproate (VPA) is an effective, widely used antiepileptic drug. Unfortunately its use in pregnant women is associated with neural tube defects in the offspring. Although the etiology of neural tube defects is multifactorial, there is evidence that underlying genetic susceptibility plays a part. We describe two women taking moderate doses of VPA who repeatedly bore children with neural tube defects, despite folate supplementation. This suggests a pharmacogenetic susceptibility to the teratogenic effects of VPA. PMID- 11422331 TI - Incidence of drug-induced aggravation in benign epilepsy with centrotemporal spikes. AB - PURPOSE: Benign epilepsy with centrotemporal spikes (BECTS) is characterized by an excellent prognosis. Drug therapy is necessary in only a minority of patients. Carbamazepine (CBZ) and phenobarbital (PB) have been reported to cause electroclinical aggravation in some cases. The incidence of drug-induced aggravation in BECTS has never been established. METHODS: We retrospectively studied 98 consecutive cases of BECTS, examined at the Centre Saint Paul between 1984 and 1999; 82 patients had received one or more treatments, often successively and in association. RESULTS: We found only one case of electroclinical aggravation with CBZ among 40 patients exposed to CBZ (35 in monotherapy, five in polytherapy). An additional case showed a marked EEG aggravation on CBZ + PB among 14 patients taking PB (nine with monotherapy and five with polytherapy), and PB was apparently responsible. No patient treated with valproate or benzodiazepines showed aggravation. CONCLUSIONS: Aggravation of BECTS caused by antiepileptic drugs happens only rarely. There is a minor risk of aggravation with CBZ and also probably with PB. Drug-induced aggravation may occur only during certain periods coinciding with spontaneous worsening of BECTS. PMID- 11422332 TI - Prognostic significance of failure of the initial antiepileptic drug in children with absence epilepsy. AB - PURPOSE: In children with childhood absence epilepsy (CAE) and juvenile absence epilepsy (JAE), to determine the impact of failure of initial antiepileptic drug (AED) for lack of efficacy in eventual seizure control and long-term remission of epilepsy. METHODS: Centralized EEG records for the province of Nova Scotia allowed identification of all children seen with CAE or JAE between 1977 and 1985. Information regarding success or failure of initial AED in fully controlling seizures and long-term seizure control and remission of epilepsy was collected by patient questionnaire and chart review. RESULTS: Eighty-six of 92 eligible patients were followed up (75 CAE, 11 JAE). Initial AED treatment was successful in 52 (60%) of 86. Success tended to be greater for valproate (VPA) than for other AEDs (p = 0.07), and lower if generalized tonic-clonic or myoclonic seizures coexisted (p < 0.004 and p < 0.03). Terminal remission was more likely if the initial AED was successful than if it had failed (69% vs. 41%; p < 0.02). Compared with those in whom the initial AED was successful, subjects whose initial AED had failed were more likely to progress to juvenile myoclonic epilepsy (JME) at last follow-up (32% vs. 10%; p < 0.02) and to develop intractable epilepsy (17% vs. 2%; p < 0.04). CONCLUSIONS: Initial AED was successful in 60% of children with AE. If the first AED failed, the outcome was less favorable, with a lower rate of terminal remission and a higher rate of progression to JME and intractable epilepsy. PMID- 11422333 TI - Comparative cognitive effects of carbamazepine and gabapentin in healthy senior adults. AB - PURPOSE: This study compared the cognitive effects of carbamazepine (CBZ) and gabapentin (GBP) in healthy senior adults by using a randomized, double-blind crossover design. METHODS: Thirty-four senior adults were randomized to receive one of the two drugs followed by a 5-week treatment period. A 4-week washout phase preceded initiation of the second drug. Antiepileptic drugs (AEDs) were titrated to target doses of either CBZ (800 mg/day) or GBP (2,400 mg/day). Primary outcome measures were standardized neuropsychological tests of attention/vigilance, psychomotor speed, motor speed, verbal and visual memory, and the Profile of Mood State (POMS), yielding a total of 17 variables. Each subject received cognitive testing at predrug baseline, end of first drug phase, end of second drug phase, and 4 weeks after completion of the second drug phase. RESULTS: Fifteen senior adults (mean age, 66.5 years; range, 59-76 years) completed the study. Seniors completing the study did not differ significantly from noncompleting seniors in terms of demographic features or baseline cognitive performances. Fifteen of the 19 seniors not completing the study dropped out while receiving CBZ. Adverse events were frequently reported for both AEDs, although they were more common for CBZ. Mean serum levels for the completers were within midrange clinical doses (CBZ, 6.8 microg/ml; GBP, 7.1 microg/ml). Significant differences between CBZ and GBP were found for only one of 11 cognitive variables, with better attention/vigilance for GBP, although the effect was modest. Performances on the nondrug average were significantly better on 45% of cognitive variables compared with CBZ and 36% compared with GBP. The overall pattern of means favored GBP over CBZ on 15 of 17 (p < 0.001), nondrug over CBZ on 17 of 17 (p < 0.0000), and nondrug over GBP on eight of 17 (NS). CONCLUSIONS: Mild cognitive effects were found for both AEDs compared with the nondrug average condition. The magnitude of difference between the two AEDs across the cognitive variables was modest. Self-reported mood was not significantly affected by either AED. However, overall tolerability and side-effect profile of CBZ were poorer than those of GBP in senior adults at doses and titration rates reported in this study. PMID- 11422334 TI - Utility of the scalp-recorded ictal EEG in childhood epilepsy. AB - PURPOSE: To evaluate the usefulness of the scalp-recorded ictal EEGs in diagnosing childhood epilepsy. METHODS: We analyzed the ictal EEGs of 259 seizures in 183 patients who visited the department of child neurology, Okayama University Medical School, during the past 6 years. RESULTS: We divided all seizures into the following four categories, according to the diagnostic usefulness of ictal EEGs in determining the seizure type: 1. (a) Ictal EEGs confirmed the diagnosis of the seizure type based on seizure symptoms (101 seizures); (b) Ictal EEGs aided in the classification of the seizure type based on the seizure symptoms (101 seizures); (c) Ictal EEGs corrected errors in the classification (37 seizures); and (d) Ictal EEGs revealed previously unreported/undocumented seizure type (20 seizures). 2. Of the 37 misdiagnosed seizures (group C), 11 were nonepileptic seizures misdiagnosed as epileptic seizures, eight were complex partial seizures (CPS) misdiagnosed as the other seizure types, and 10 were other seizure types misdiagnosed as CPSs. 3. Of the 20 previously unreported/undocumented seizures (group D), nine were myoclonic seizures, five were absence seizures, five were CPS, and one was tonic spasms. 4. Seventy-two patients had CPS. Among them, 11 patients showed no epileptic spikes in their interictal EEG recordings. Therefore, ictal recordings confirmed the diagnosis of epilepsy. CONCLUSIONS: Ictal EEG recording is a very useful diagnostic tool not only for determining seizure types, but also for uncovering the existence of the unsuspected seizure types. It supplies the physician with useful information for the classification and the treatment of epilepsy. In particular, ictal EEGs are useful in diagnosing patients with CPS. PMID- 11422335 TI - The absence of interictal spikes with documented seizures suggests extratemporal epilepsy. AB - PURPOSE: To determine the demographic and clinical characteristics of patients who have documented epileptic seizures on long-term video-EEG monitoring who do not have interictal spikes. METHODS: The records of 1,223 monitoring studies from 919 patients who underwent noninvasive long-term video-EEG monitoring were reviewed. In 28 patients (3.0% of monitored patients, 4.4% of patients with electrographic evidence of epilepsy), no interictal spikes were found despite the occurrence of at least one recorded electrographic seizure. The demographic, medical, neuropsychological, and EEG data of these patients were compared with those of 28 matched control patients with documented interictal spikes. RESULTS: Extratemporal seizures were significantly more frequent in the patients with at least one recorded epileptic seizure but without interictal spikes compared with patients with epileptic seizures and interictal spikes (p = 0.031). The only other significant difference between the groups (p = 0.016) was a later age at seizure onset (18.3 vs. 10.7 years) for the patients without interictal spikes. Age at evaluation, gender, handedness, clinical seizure type, family history of epilepsy, history of febrile seizures, neuropsychological testing, and neurologic and psychiatric history did not differ between the two groups. CONCLUSIONS: In patients with documented epilepsy without interictal spikes on EEG monitoring, the possibility of an extratemporal focus should be considered. PMID- 11422336 TI - Lack of association between an interleukin 1 beta (IL-1beta) gene variation and refractory temporal lobe epilepsy. AB - PURPOSE: We attempted to confirm recent findings of Kanemoto et al. that demonstrated a positive association (p < 0.017) between a polymorphism in the promoter region of the interleukin 1-beta (IL-1beta) gene and the clinical phenotype of temporal lobe epilepsy with hippocampal sclerosis (TLE+HS). METHODS: We determined the frequency of this polymorphism in a group of 61 TLE+HS patients of European ancestry and compared it with that found in 119 ethnically matched control subjects. RESULTS: Analysis of genotype and allele frequencies showed no statistically significant difference in the distribution of the polymorphism between the two groups (p = 0.10). CONCLUSIONS: These data suggest that this IL 1beta promoter polymorphism does not act as a strong susceptibility factor for TLE+HS in a population of individuals of European ancestry. PMID- 11422337 TI - The difference in perceptions of educational need between epilepsy patients and medical personnel. AB - PURPOSE: This study was undertaken to determine what patients with epilepsy need to know about their condition, and to discern what differences exist between patients' perceptions of this need and the medical profession's perception of what patients should know. METHODS: Seventy-five consecutive patients with epilepsy and 56 medical personnel (residents and nurses) who were working in either Neurology or Neurosurgery Units were studied using a structured questionnaire consisting of 3 subsets with a total of 27 questions. RESULTS: Using a Likert scale, epilepsy patients gave high priority to their need for more information about "how epilepsy is diagnosed," "the structure of the brain" (p < 0.05, p < 0.01, respectively), and "the diet that might prevent the attack" (p < 0.05) than did medical personnel. Our study also revealed that men were more concerned with questions regarding smoking and drinking than were women (p < 0.05, respectively), whereas married patients gave higher scores in the category of employment (p < 0.01) and contraception/pregnancy (p < 0.05) than did unmarried patients. The patients with one or more seizures per month rated higher scores on questions concerning the first aid at the time of attack and dosage of antiepileptic drugs (AEDs; p < 0.05, respectively) than did the patients with fewer than one seizure per month. The patients taking poly-AED treatment gave higher scores on six items related to AED therapy than did those receiving monotherapy. No significant differences were found with regard to the length of time patients had the condition, nor to educational level or current employment status. CONCLUSIONS: An educational program for epilepsy patients should be developed on the basis of understanding that there are differences in perspectives among patients with different sociocultural contexts as well as between patients and medical personnel. PMID- 11422338 TI - Myoclonus in epilepsy patients with anticonvulsive add-on therapy with pregabalin. AB - PURPOSE: To report on the occurrence of myoclonus in patients receiving pregabalin (PGB) for the treatment of focal epilepsy. METHODS: Clinic records of 19 patients who were consecutively enrolled at a tertial referral epilepsy center in a randomized, double-blind and/or open add-on study with PGB were reviewed. RESULTS: In four patients treated with PGB, focal myoclonus newly developed. The side effect appeared with PGB doses of 50-600 mg/day; the intensity showed some dose dependency. All patients had medically refractory focal epilepsy and received other antiepileptic drugs (AEDs) besides the study medication. One patient showed focal myoclonic jerks of the left arm, whereas the other patients developed multifocal myoclonus. Polygraphic studies including electromyogram (EMG)-triggered back-averaging of the EEG in the patient with the highest frequency of myoclonic jerks showed no visible correlate of the myoclonus. In this patient, frequency and intensity of myoclonic jerks significantly decreased after dose reduction of PGB. In the other cases, myoclonus was only subtle and did not significantly interfere with daily activities, so that a dose reduction of PGB was not considered necessary. CONCLUSIONS: These data indicate a relatively high incidence (four of 19) of myoclonus associated with PGB therapy. The rate seems to be at least as high as reported in patients receiving the structurally similar anticonvulsant gabapentin. PMID- 11422339 TI - Oxcarbazepine in focal epilepsy and hepatic porphyria: a case report. AB - PURPOSE: Despite the development of new antiepileptic agents (AEDs), the therapy of epilepsies along with hepatic porphyrias remains difficult. Most AEDs such as carbamazepine (CBZ), phenytoin (PHT), valproate (VPA), and lamotrigine (LTG) may precipitate clinically latent porphyria by inducing hepatic metabolism and increasing hepatic heme synthesis. Actually, only gabapentin (GBP), an AED without any hepatic metabolism, is known as a potential therapy for partial seizures in patients having hepatic forms of porphyria. METHODS: We present the case of a 28-year-old man with porphyria cutanea tarda (PCT) who has had pharmacoresistant epilepsy with complex partial and secondarily generalized seizures since early childhood. Despite having undergone several AED therapies over the years, no seizure-free interval had been observed. Only CBZ could cause a seizure reduction, but this treatment had to be discontinued as an elevation of the transaminases as well as pruritus and erythema were noted. The patient was then started on oxcarbazepine (OCBZ), a ketoanalogue of CBZ similar in its pharmacologic mechanism as well as its clinical use, but which, in contrast to CBZ, has only a low hepatic induction of microsomal enzymes. A final maintenance dose four times higher than that of CBZ was prescribed. RESULTS: In the follow up, the patient stopped having seizures, and his liver functions became normal. CONCLUSIONS: It can be concluded that OCBZ can successfully be administered to patients with hepatic porphyria and focal epilepsy who did not respond to treatment with GBP. PMID- 11422340 TI - A proposed diagnostic scheme for people with epileptic seizures and with epilepsy: report of the ILAE Task Force on Classification and Terminology. PMID- 11422341 TI - Guidelines for essential services, personnel, and facilities in specialized epilepsy centers in the United States. PMID- 11422343 TI - Learning disability in epilepsy: definitions and classification. PMID- 11422344 TI - The epidemiology of epilepsy and learning disability. PMID- 11422345 TI - Proposal for a multicenter study on epilepsy and learning disorders in children. PMID- 11422346 TI - Pathogenesis of learning disabilities in epilepsy. PMID- 11422347 TI - Cognitive performance, subtle seizures, and the EEG. PMID- 11422349 TI - Learning disabilities and language function in epilepsy. PMID- 11422350 TI - Chromosome abnormalities and epilepsy. PMID- 11422352 TI - Depression and anxiety in patients with epilepsy. PMID- 11422353 TI - Measurement and testing in adults. PMID- 11422355 TI - Generalized versus selective cognitive impairments resulting from brain damage sustained in childhood. PMID- 11422356 TI - Psychological and neuropsychological assessment before and after surgery for epilepsy: implications for the management of learning-disabled people. PMID- 11422357 TI - Evidence-based prescribing in adults with learning disability and epilepsy. PMID- 11422358 TI - Effects of antiepileptic drugs on cognition. PMID- 11422360 TI - Treatment of state-dependent learning disability. PMID- 11422361 TI - Multidisciplinary and multiagency contributions to care for those with learning disability who have epilepsy. PMID- 11422363 TI - Epilepsy and learning disabilities:closing comments. PMID- 11422364 TI - Purified components of the Escherichia coli Tat protein transport system form a double-layered ring structure. AB - The Escherichia coli twin arginine translocation (Tat) system mediates Sec independent export of protein precursors bearing twin arginine signal peptides. The genes tatA, tatB, tatC and tatE code for integral membrane proteins that are components of the Tat pathway. Cells co-overexpressing tatABCDE show an increased rate of export of a signal peptide-defective Tat precursor protein and a complex containing the TatA and TatB proteins can be purified from the membranes of such cells. The purified TatAB complex has an apparent molecular mass of 600 kDa as measured by gel permeation chromatography and, like the membranes of wild-type cells, contains a large molar excess of TatA over TatB. Negative stain electron microscopy of the complex reveals cylindrical structures that may correspond to the Tat protein transport channel. PMID- 11422365 TI - C-Terminal region of human NAD+-dependent 15-hydroxyprostaglandin dehydrogenase is involved in the interaction with prostaglandin substrates. AB - NAD+-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH) catalyzes the oxidation of the 15(S) hydroxyl group of prostaglandins to a 15-keto group resulting in a significant reduction of the biological activities of prostaglandins. Although the key residues involved in NAD+ binding and in catalytic activity have been partially identified, the sites of interaction of the enzyme with the prostaglandin substrates are yet to be determined. Homology analysis of the primary structures of 15-PGDH from human, mouse and rat indicates that the sequences are almost homologous except for two regions near the C terminus. The involvement of the C-terminal region in catalytic activity was examined by studies on C-terminally truncated enzymes and on human/rat chimeric enzymes. When three to four amino acids were removed successively from the C terminal end of human 15-PGDH, the truncated enzymes exhibited decreasing Vmax/Km ratios and increasing Km values for PGE2 as the chain was shortened. Similarly, when the C-terminal 14 amino acids of human 15-PGDH were replaced by the C terminal 14 amino acids of rat 15-PGDH or vice versa, the Vmax/Km ratios and the Km values for prostaglandin E2 of the chimeric enzymes were in between those of the two wild-type enzymes. This indicates that the catalytic effectiveness of human 15-PGDH decreases as the C-terminal region is gradually removed or replaced by rat sequences. The C-terminal region appears to be more important for the interaction of the enzyme with the prostaglandin substrates than with the coenzyme. PMID- 11422366 TI - Methionine oxidation and its effect on the stability of a reconstituted subunit of the light-harvesting complex from Rhodospirillum rubrum. AB - An additional component in the purified core light-harvesting complex (LH1) from wild-type purple photosynthetic bacterium Rhodospirillum rubrum has been identified as an oxidized species of alpha-polypeptide by MALDI-TOF mass spectrometry. This component appears as a slightly earlier-eluting peak in the RP HPLC chromatogram compared with the authentic alpha-polypeptide. The oxidation site has been determined to be the N-terminal methionine residue by high resolution NMR spectroscopy, where the methionine is oxidized to methionine sulfoxide in a diastereoisomeric form. Interconversion between the oxidized and authentic alpha-polypeptides has been confirmed by selective oxidation and reduction. The oxidative modification of methionine is shown to have discernible effects on the ability to form B820 subunit with beta-polypeptide and bacteriochlorophyll a, and on the stability of the reconstituted B820 subunit. Both the ability and the stability for the samples using the oxidized alpha polypeptide are moderately reduced, indicating that the oxidation-induced conformational change in the N-terminal domain of alpha-polypeptide may affect the pigment-binding environment through a long-range interaction. The MALDI-TOF mass results also reveal that the N-terminus of alpha-polypeptide is formylated and no phosphorylation has occurred in this polypeptide. PMID- 11422367 TI - Phosphorylation of proteins in the light-dependent signalling pathway of a filamentous cyanobacterium. AB - The genome of the filamentous cyanobacterium Calothrix sp. PCC7601 contains two genes, cphA and cphB, encoding proteins with similarity to plant phytochromes and bacterial histidine kinases. In vitro, CphA and CphB readily attach a tetrapyrrole chromophore to develop spectrally active holoproteins that are photointerconvertible between a red light-absorbing and a far-red light-absorbing form. Together with the putative response regulators, RcpA and RcpB, the putative histidine kinases, CphA and CphB, are suggested to constitute two two-component systems of light-dependent signal transduction. In this report, we demonstrate the kinase activity of both CphA and CphB. In vitro experiments carried out on the purified proteins show that CphA and CphB are autophosphorylated in the presence of ATP and that phospho-CphA is capable of efficient phosphotransfer to RcpA as is phospho-CphB towards RcpB. The autophosphorylation and the phosphorelay are dependent on light. Both activities are reduced under red light vs. far-red light irradiation. No phosphoryl transfer occurred between phospho CphA and RcpB or between phospho-CphB and RcpA. The response regulators RcpA and RcpB can receive a phosphoryl moiety also from the small phospho-donor acetyl phosphate. The stability of the phosphorylated regulators is not affected by CphA and CphB or light. PMID- 11422368 TI - Organization of the genes involved in dimethylglycine and sarcosine degradation in Arthrobacter spp.: implications for glycine betaine catabolism. AB - The nucleotide sequences of two cloned DNA fragments containing the structural genes of heterotetrameric sarcosine oxidase (soxBDAG) and dimethylglycine dehydrogenase (dmg) from Arthrobater spp. 1-IN and Arthrobacter globiformis, respectively, have been determined. Open reading frames were identified in the soxBDAG operon corresponding to the four subunits of heterotetrameric sarcosine oxidase by comparison with the N-terminal amino-acid sequences and the subunit relative molecular masses of the purified enzyme. Alignment of the deduced sarcosine oxidase amino-acid sequence with amino-acid sequences of functionally related proteins indicated that the arthrobacterial enzyme is highly homologous to sarcosine oxidase from Corynebacterium P-1. Deletion and expression analysis, and alignment of the deduced amino-acid sequence of the dmg gene, showed that dmg encodes a novel dimethylglycine oxidase, which is related to eukaryotic dimethylglycine dehydrogenase, and contains nucleotide-binding, flavinylation and folate-binding motifs. The recombinant dimethylglycine oxidase was purified to homogeneity and characterized. The DNA located upstream and downstream of both the soxBDAG and dmg genes is predicted to encode enzymes involved in the tetrahydrofolate-dependent assimilation of methyl groups. Based on the sequence analysis reported herein, pathways are proposed for glycine betaine catabolism in Arthrobacter species, which involve the identified folate-dependent enzymes. PMID- 11422369 TI - Synthesis of novel anti-inflammatory peptides derived from the amino-acid sequence of the bioactive protein SV-IV. AB - SV-IV is a basic, thermostable, secretory protein of low Mr (9758) that is synthesized by rat seminal vesicle (SV) epithelium under strict androgen transcriptional control. This protein is of obvious pharmacological interest because it has potent nonspecies-specific immunomodulatory, anti-inflammatory, and pro-coagulant activities. In evaluating the clinical relevance and the possible use in medicine of SV-IV, we became interested in the study of its structure-function relationships and aimed to identify in its polypeptide chain specific peptide fragments possessing the marked anti-inflammatory properties of the protein not associated with other biological activities (pro-coagulation and immunomodulation) typical of this molecule. By using two different experimental approaches (the fragmentation of the protein into peptide derivatives by chemical methods and the organic synthesis on solid phase of selected peptide fragments), data were obtained showing that in this protein: (a) the immunomodulatory activity is related to the structural integrity of the whole molecule; (b) the anti-inflammatory activity is located in the N-terminal region of the molecule, the 8-16 peptide fragment being the most active; (c) the identified anti inflammatory peptide derivatives do not seem to possess pro-coagulant activity, even though this particular function has been located in the 1-70 segment of the molecule. PMID- 11422370 TI - Generation and epitope mapping of high-affinity scFv to eukaryotic elongation factor 1A by dual application of phage display. AB - To generate specific tools for, in particular, localization studies of the eukaryotic elongation factor 1A (eEF1A), we have applied phage display in various formats to affinity-improve and map epitopes of two previously isolated, low affinity single-chain Fv (scFv) G3 and D1. The scFv differ in their reactivity toward the eEF1A isoforms, eEF1A-1 and eEF1A-2. By PCR-based randomization of six residues within the variable light chain CDR3 (LCDR3), and subsequent phage-based affinity-selection, two 'families' of affinity-improved scFv were obtained. The scFv of highest affinity, A8, has a Kd of 9 nM to eEF1A-1. Interestingly, two affinity-improved scFvs have abnormally short LCDR3 consisting of two and four residues compared to 11 in the parental scFv. Hence, the LCDR3 of the parental clones may play a modulating rather than a direct role in antigen-binding. Despite different preferences for the eEF1A isoforms, both families of scFv recognize antigenic determinant(s), which was mapped to residues 413-450 of eEF1A 1/2 by Western blot analysis of recombinant human eEF1A (hEF1A) fragments. Prior to the Western blotting analysis, the epitope location had been suggested using a novel approach where phage-antibody repertoire derived scFv were used to select phage-displayed peptides. Hereby, peptides containing a SFXD motif, matching the SFSD(414-418) sequence found in hEF1A-1 were isolated. The structure of eukaryotic EF1A from yeast indicates a discontinuous nature of the epitope with distal functional elements juxtaposed by the protein fold. Finally, the scFv A8 was applied for immunofluorescence studies of transformed human amnion cells and MCF-7 fibroblasts. In both cases a perinuclear localization of hEF1A was observed. No evidence for the reported nuclear localization of hEF1A was obtained. PMID- 11422371 TI - The ABC transporter Pdr5p mediates the efflux of nonsteroidal ecdysone agonists in Saccharomyces cerevisiae. AB - We have previously shown that the synthetic nonsteroidal ecdysone agonist tebufenozide (RH-5992) is actively excluded by resistant cells of insects. To identify the transporter that could be involved in the efflux of RH-5992, the role of three ATP binding cassette transporters, Pdr5p, Snq2p and Ycf1p, has been studied using transporter-deletion mutants of yeast Saccharomyces cerevisiae. PDR5 (pleiotropic drug resistance 5) deletion mutants (Deltapdr5 and Deltapdr5Deltasnq2) retained significantly higher levels of 14C-radiolabeled RH 5992 within the cells when compared to wild-type strain or single deletion mutants of SNQ2 (Deltasnq2) and YCF1 (Deltaycf1). Introduction of an expression vector containing the PDR5 gene into the PDR5 single deletion mutant reversed the effect, resulting in the active exclusion of [14C]RH-5992 from these cells as efficiently as the wild-type cells. These results demonstrated that the ABC transporter Pdr5p but not Snq2p or Ycf1p was responsible for the active exclusion of [14C]RH-5992 in yeast. This exclusion was temperature-dependent and was blocked by the ATPase inhibitors oligomycin and vanadate, indicating that the efflux was an active process. The mutants with the PDR5 deletion can also selectively accumulate [14C]RH-0345 and [14C]RH-2485, but not [14C]RH-5849, indicating that these three compounds share the same transporter Pdr5p for efflux. PMID- 11422372 TI - Proteolytic processing of a human salivary proline-rich protein precursor by proprotein convertases. AB - Salivary proline-rich proteins (PRPs) are synthesized as precursors that are cleaved before secretion giving rise to glycosylated PRPs which have lubricating function and basic PRPs which are potent precipitators of dietary tannins. The putative cleavage sites in the precursors for basic and glycosylated PRPs all conform to the sequence RSXR downward arrowS (X can be A, S or P) in agreement with the recognition sequence (RXXR downward arrow) for various proprotein convertases. PRB4S, a proprotein giving rise to a basic PRP (IB-5) as well as a glycosylated PRP (II-1) was synthesized by in vitro transcription-translation. It was cleaved by furin at RSAR downward arrowS(173-178) giving rise to two proteins II-1 and IB-5. Similarly another precursor with the sequence RSAR downward arrowS(173-178) was also cleaved by furin. This together with previous results show that in vitro furin can cleave all RSXR downward arrowS sequences in the proproteins that give rise to glycosylated and basic PRPs. To demonstrate cellular cleavage, a human submandibular cell line (HSG) was transfected with a vector encoding PRB4S. This resulted in secretion of II-1 and IB-5. The degree of cleavage was enhanced by coexpressing furin and PRB4S. No cleavage occurred if the cells expressed a mutant PRB4S, R177Q, where the furin cleavage site had been destroyed. Cleavage was also inhibited if a furin inhibitor was coexpressed with PRB4S. Incubating the cells at 20 degrees C which blocks exit of proteins from the trans-Golgi network demonstrated that cleavage occurs before exit of the proteins from this network. These results show that furin may be responsible for in vivo cleavage of PRP precursors. Transfecting furin-deficient RPE.40 cells with a vector encoding PRB4S also led to secretion of II-1 and IB-5 showing that convertases other than furin can also cleave PRB4S in tissue culture. PMID- 11422373 TI - Samui, a novel cold-inducible gene, encoding a protein with a BAG domain similar to silencer of death domains (SODD/BAG-4), isolated from Bombyx diapause eggs. AB - Cellular responses to cold-acclimation have not yet been studied in depth. To explore this field, we focussed on insect diapause development. Although embryonic diapause of Bombyx mori is sustained at 25 degrees C, chilling at 5 degrees C for 2 months causes diapause termination, a transition that is marked when the sorbitol dehydrogenase gene (SDH) is activated. To clarify the relationship between this activation and incubation at 5 degrees C, we isolated a novel cold-inducible gene, Samui. Expression of Samui mRNA and protein was activated after incubation at 5 degrees C for 5-6 days, lasted for another 30 days and then weakened. Exposure to 25 degrees C suppressed both mRNA and protein expression. In nondiapause eggs incubated at 5 degrees C, Samui was also up regulated, although the expression was weaker. Samui contained nuclear localization-signals, a ssDNA-binding motif and a BAG domain similar to that of SODD/BAG-4. Because Samui could bind to HSP70, it is a member of BAG protein family. It is proposed that Samui serves to transmit the '5 degrees C signal' for SDH expression in diapause eggs, while also protecting against cold-injures in nondiapause eggs, through binding to respective partners. This is the first report that a member of BAG protein family is up-regulated by cold. PMID- 11422374 TI - Oxidative and hydrolytic properties of beta-amyloid. AB - beta-Amyloid protein is the major component of senile plaques found in the brains of Alzheimer's patients. Previously, a new biochemical property of amyloid, its ability to disrupt ester and peptide bonds, was described [Elbaum, D., Brzyska, M., Bacia, A. & Alkon, D. (2000) Biochem. Biophys. Res. Commun. 267, 733-738]. In the present work we compare the ability of beta-amyloid to hydrolyse and oxidize model fluorescent derivatives of dichlorofluorescein [dichlorodihydrofluorescein (H2DCF) or dichlorofluorescein diacetate (DCF-DA), respectively] to the same final product (dichlorofluorescein). Although there is accumulating evidence of oxidative properties of beta-amyloid, little is known about its hydrolytic abilities. Chemical modification studies revealed that hydrolytic properties are related to a His, Ser and Asp/Glu triad, while residues of His, Tyr and Met are involved in the oxidative activity of amyloid. Studies with the rat homologue of human beta-amyloid (1-40), containing three amino-acid substitutions (Arg5-->Gly, Tyr10-->Phe and His13-->Arg) confirmed a role of His in the studied processes. Reduction of the hydrolysis product caused by inhibitors of Ser esterases (phenylmethylsulphonyl fluoride and eserine) suggests that beta-amyloid-mediated hydrolysis is Ser sensitive. Antioxidants and metal chelators that reduced H2DCF oxidation did not change or increase DCF-DA hydrolysis. Solvent isotope effects suggest the involvement of hydrogen bonds in the hydrolysis reaction. Hydrolysis was inhibited by redox-active metal ions and was practically oxygen independent while the oxidation process was redox-active-metal enhanced [Cu(II) and Fe(II) primarily], and oxygen dependent. Product formation was significantly inhibited by catalase and superoxide dismutase as well as benzoquinone, a specific superoxide anion radical scavenger. Increase of fluorescence by oxidation was strongly inhibited by azide and His and enhanced in samples prepared with deuterated phosphate buffer, suggesting singlet oxygen intermediacy. Our data are consistent with superoxide-mediated singlet oxygen intermediate in this Fenton mechanism-driven reaction. These results indicate that hydrolytic and oxidative properties of beta-amyloid are distinct features of this peptide and probably require different mechanisms to occur, but both of them may contribute to beta amyloid toxicity. PMID- 11422375 TI - Sequence-dependent binding of bis-amidine carbazole dications to DNA. AB - The conventional wisdom argues that DNA intercalators possess a condensed polyaromatic ring whereas DNA minor groove binders generally contain unfused aromatic heterocycles, frequently separated by amide bonds. Recently, this view has been challenged with the discovery of powerful intercalating agents formed by unfused aromatic molecules and groove binders containing a polyaromatic nucleus. Bis-amidinocarbazoles belong to this later category of drugs having a planar chromophore and capable of reading the genetic information accessible within the minor groove of AT-rich sequences [Tanious, F.A., Ding, D., Patrick, D.A., Bailly, C., Tidwell, R.R. & Wilson, W.D. (2000) Biochemistry 39, 12091-12101]. But in addition to the tight binding to AT sites, we show here that bis amidinocarbazoles can also interact with GC sites. The extent and mode of binding of 2,7 and 3,6 substituted amidinocarbazoles to AT and GC sequences were investigated by complementary biochemical and biophysical methods. Absorption, fluorescence, melting temperature and surface plasmon resonance (SPR) measurements indicate that the position of the two amidine groups on the carbazole ring influences significantly the drug-DNA interaction. SPR and DNase I footprinting data confirm the AT-preference of the compounds and provide useful information on their additional interaction with GC sequences. The 3,6-carbazole binds approximately twice as strongly to the GC-containing hairpin oligomer than the 2,7-regioisomer. The high tendency of the 3,6 compound to intercalate into different types of DNA containing G.C base pairs is shown by electric linear dichroism. This work completes our understanding of the sequence-dependent DNA binding properties of carbazole dications. PMID- 11422376 TI - The effect of nucleotides and mitochondrial chaperonin 10 on the structure and chaperone activity of mitochondrial chaperonin 60. AB - Mitochondrial chaperonins are necessary for the folding of newly imported and stress-denatured mitochondrial proteins. The goal of this study was to investigate the structure and function of the mammalian mitochondrial chaperonin system. We present evidence that the 60 kDa chaperonin (mt-cpn60) exists in solution in dynamic equilibrium between monomers, heptameric single rings and double-ringed tetradecamers. In the presence of ATP and the 10 kDa cochaperonin (mt-cpn10), the formation of a double ring is favored. ADP at very high concentrations does not inhibit malate dehydrogenase refolding or ATP hydrolysis by mt-cpn60 in the presence of mt-cpn10. We propose that the cis (mt cpn60)14.nucleotide.(mt-cpn10)7 complex is not a stable species and does not bind ADP effectively at its trans binding site. PMID- 11422377 TI - Characterization of human muscle type cofilin (CFL2) in normal and regenerating muscle. AB - Cofilins are actin binding proteins and regulate actin assembly in vivo. Numerous cofilin homologues have been characterized in various organisms including mammals. In mice, a ubiquitously expressed cofilin (CFL1) and a skeletal muscle specific cofilin (CFL2) have been described. In the present study, we identified and characterized a human CFL2 gene localized on chromosome 14, with high homology to murine CFL2. Furthermore, we provide evidence for differentially spliced CFL2 transcripts (CFL2a and CFL2b). CFL2b is expressed predominantly in human skeletal muscle and heart, while CFL2a is expressed in various tissues. Genetic defects of CFL2 were excluded for one human muscle disorder, the chromosome 14 linked distal myopathy MPD1, and shown to be only possible to be a rare cause of another, nemaline myopathy. In a mouse model of mechanically induced muscle damage the changes of cofilin expression were monitored during the first 10 days of regeneration, with dephosphorylated CFL2 being the major isoform at later stages of muscle regeneration. A similar predominance of dephosphorylated CFL2 was observed in chronically regenerating dystrophin deficient muscles of Duchenne muscular dystrophy patients. Therefore, the CFL2 isoform may play an important role in normal muscle function and muscle regeneration. PMID- 11422378 TI - Characterization of a new brain-specific isoform of the EWS oncoprotein. AB - EWS and related TAFII68 and TLS/FUS genes are fused with different genes encoding transcription factors in various human cancers. The products of these genes have the ability to bind RNA and have been shown to be part of splicing and transcription complexes. We show that the EWS, TAFII68 and TLS/FUS proteins are expressed to various levels in all adult murine tissues. We characterize a new isoform of EWS that is specifically expressed in the central nervous system, in both mice and humans. It is shown to be related to a splice variant which includes a new 18-bp exon, termed 4', between exon 4 and 5. The detection of this isoform in spontaneously differentiating SH-SY5Y neuroblastoma cells and in nerve growth factor-induced PC12 cells further links this isoform to neural differentiation. RT-PCR experiments indicate that the level of expression of the brain-specific EWS isoform is stable during brain development whereas that of the ubiquitous EWS isoform decreases during this period. The two isoforms show a parallel decrease in expression after birth. The 4' exon is not detected in tumour-specific EWS fusion transcripts, suggesting that its presence may impair their oncogenic properties. Interestingly, sequences of the 4' exon and flanking regions show remarkable similarities to that of the neural-specific c-src exon, suggesting common mechanisms for the alternative splicing of these exons. The phylogenetic conservation and relationship to neural differentiation strongly suggests an important functional role for this exon. PMID- 11422379 TI - Fibrate induction of the adrenoleukodystrophy-related gene (ABCD2): promoter analysis and role of the peroxisome proliferator-activated receptor PPARalpha. AB - X-linked adrenoleukodystrophy (X-ALD) is a neurodegenerative disease due to a defect in the ABCD1 (ALD) gene. ABCD1, and the two close homologues ABCD2 (ALDR) and ABCD3 (PMP70), are genes encoding ATP-binding cassette half-transporters of the peroxisomal membrane. As overexpression of the ABCD2 or ABCD3 gene can reverse the biochemical phenotype of X-ALD (reduced beta-oxidation of very-long chain fatty acids), pharmacological induction of these partially redundant genes may represent a therapeutic approach to X-ALD. We previously reported that the ABCD2 and ABCD3 genes could be strongly induced by fibrates, which are hypolipidaemic drugs and peroxisome-proliferators in rodents. We provide evidence that the induction is dependent on peroxisome proliferator-activated receptor (PPARalpha) as both genes were not induced in fenofibrate-treated PPARalpha -/- knock-out mice. To further characterize the PPARalpha pathway, we cloned and analysed the promoter of the ABCD2 gene, the closest homologue of the ABCD1 gene. The proximal region (2 kb) of the rat promoter displayed a high conservation with the human and mouse cognate sequences suggesting an important role of the region in regulation of the ABCD2 gene. Classically, fibrate-induction involves interaction of PPARalpha with a response element (PPRE) characterized by a direct repeat of the AGGTCA-like motif. Putative PPRE motifs of the rat ABCD2 promoter were studied in the isolated form or in their promoter context by gel-shift assay and transfection of COS-7 cells. We failed to characterize a functional PPRE, suggesting a different mechanism for the PPARalpha-dependent regulation of the ABCD2 gene. PMID- 11422380 TI - Transfection of the c-erbB2/neu gene upregulates the expression of sialyl Lewis X, alpha1,3-fucosyltransferase VII, and metastatic potential in a human hepatocarcinoma cell line. AB - The pCMV4 plasmid containing the cancer-promoting gene, c-erbB2/neu, was cotransfected into the human hepatocarcinoma cell line 7721 with the pcDNA3 vector, which contains the 'neo' selectable marker. Several clones showing stable expression of c-erbB2/neu were established and characterized by determination of c-erbB2/neu mRNA and its encoded protein p185. Expression of Lewis antigens and alpha1,3-fucosyltransferases and the biological behavior of 7721 cells after c erbB2/neu transfection were studied using mock cells transfected with the vectors pCMV4 and pcDNA3 as controls. SLe(x) expression on the surface of mock cells was high, whereas expression of SDLe(x), Lex and SLe(a) was absent or negligible. This is compatible with the abundant expression of alpha1,3-fucosyltransferase VII, very low expression of alphafucosyltransferase III/VI, and almost absent expression of alpha1,3-fucosyltransferase IV in the mock cells. After transfection of c-erbB2/neu, expression of SLe(x) and alpha1,3-fucosyltransferase VII were simultaneously elevated, but that of alphafucosyltransferase III/VI was not altered. The expression of both SLe(x) and alpha1,3-fucosyltransferase VII correlated positively with the expression of c-erbB2/neu in different clones, being highest in clone 13, medium in clone 6, and lowest in clone 7. In addition, the adhesion of 7721 cells to human umbilical vein endothelial cells (HUVECs) or P-selectin, as well as cell migration and invasion, were increased in c-erbB2/neu transfected cells. These increases also correlated positively with the expression intensities of c-erbB2/neu, SLe(x) and alpha1,3-fucosyltransferase VII in the different clones, whereas cell adhesion to fibronectin correlated negatively with these variables. mAbs to SLe(x) (KM93) and SDLe(x) (FH6) significantly and slightly, respectively, abolished cell adhesion to HUVECs or P-selectin and cell migration and invasion. mAbs to SDLe(x) and SLe(a) did not suppress cell adhesion to HUVECs nor inhibit cell migration and invasion. Transfection of alpha1,3 fucosyltransferase VII cDNA into 7721 cells showed similar results to transfection of c-erbB2/neu, and the increased adhesion to HUVECs, cell migration, and invasion were also inhibited significantly by KM93 and slightly by FH6. These results indicate that expression of alpha1,3-fucosyltransferase VII and its specific product, SLe(x), and their capacity for cell adhesion, migration and invasion are closely related. Therefore, the c-erbB2/neu gene is proposed to be a metastasis-promoting gene, and its effects are at least partially mediated by the increased expression of alpha1,3-fucosyltransferase VII and SLe(x). PMID- 11422381 TI - Platelet collagen receptor integrin alpha2beta1 activation involves differential participation of ADP-receptor subtypes P2Y1 and P2Y12 but not intracellular calcium change. AB - In agonist-induced platelet activation, the collagen platelet receptor integrin alpha2beta1 is activated to high-affinity states through ADP involvement [Jung, S.M. & Moroi, M. (2000) J. Biol. Chem. 275, 8016-8026]. Here we determined the ADP-receptor subtypes involved and their relative contributions to alpha2beta1 activation (assessed by soluble-collagen binding) using the P2Y12 antagonist AR C69931MX and P2Y1 antagonists adenosine 3',5'-diphosphate (Ado(3,5)PP) and adenosine 3'-phosphate 5'-phosphosulfate (AdoPPS). All three inhibited alpha2beta1 activation induced by low or high ADP, low thrombin, or low collagen related peptide (CRP) concentrations; however, AR-C69931MX was markedly more inhibitory than the P2Y1 antagonists, suggesting the greater contribution of P2Y12. Inhibition patterns by various combinations of AR-C69931MX, AdoPPS, and wortmannin suggested that P2Y1 and P2Y12 mediate alpha2beta1 activation through different pathways, with possible involvement of phosphoinositide 3-kinase in both. Low concentrations of the acetoxy-methyl derivative of 1,2-bis(o aminophenoxy) ethane-N,N,N',N'-tetra-acetic acid (calcium chelator) markedly decreased alpha2beta1 activation by low thrombin or CRP, but did not affect that by low or high ADP. Measurements of intracellular Ca2+ level (fluorimetric method) and alpha2beta1 activation (soluble-collagen binding) in the same platelet preparation indicated that alpha2beta1 activation via ADP receptors was independent of intracellular Ca2+ release. Our data indicate that integrin alpha2beta1 activation by ADP occurs through an inside-out signaling mechanism involving differential contributions by P2Y1 and P2Y12 wherein each contributes to some portion of the activation, with the stronger contribution of P2Y12. Furthermore, intracellular Ca2+ increase is not directly related to integrin alpha2beta1 activation, meaning that it is separate from the calcium mobilization pathways that these two ADP receptors are involved in. PMID- 11422382 TI - Human neutrophils employ the myeloperoxidase/hydrogen peroxide/chloride system to oxidatively damage apolipoprotein A-I. AB - The structural integrity of apolipoprotein A-I (apo A-I) is critical to the physiological function of high-density lipoprotein (HDL). Oxidized lipoproteins are thought to be of central importance in atherogenesis, and oxidation products characteristic of myeloperoxidase, a heme protein secreted by activated phagocytes, have been detected in human atherosclerotic tissue. At plasma concentrations of halide ion, hypochlorous acid is a major product of the myeloperoxidase-hydrogen peroxide-chloride system. We therefore investigated the effects of activated human neutrophils, a potent source of myeloperoxidase and hydrogen peroxide, on the protein and lipid components of HDL. Both free and HDL associated apo A-I exposed to activated human neutrophils underwent extensive degradation as monitored by RP-HPLC and Western blotting with a polyclonal antibody to apo A-I. Replacement of the neutrophils with reagent HOCl resulted in comparable damage (at molar oxidant : HDL subclass 3 ratio = 100) as observed in the presence of activated phagocytes. Apo A-I degradation by activated neutrophils was partially inhibited by the HOCl scavenger methionine, by the heme inhibitor azide, by chloride-free conditions, by the peroxide scavenger catalase, and by a combination of superoxide dismutase (SOD)/catalase, implicating HOCl in the cell-mediated reaction. The addition of a protease inhibitor (3,4 dichloroisocoumarin) further reduced the extent of apo A-I damage. In contrast to the protein moiety, there was little evidence for oxidation of unsaturated fatty acids or cholesterol in HDL3 exposed to activated neutrophils, suggesting that HOCl was selectively damaging apo A-I. Our observations indicate that HOCl generated by myeloperoxidase represents one pathway for protein degradation in HDL3 exposed to activated phagocytes. PMID- 11422383 TI - Changes in the levels of low-abundance brain proteins induced by kainic acid. AB - Low-abundance gene products are of interest in proteomic studies, because they are probably involved in disease-related changes and their altered levels or modifications may carry significant biological information. Detection of low abundance proteins of a proteome is one of the major limitations of proteomics and a scientific challenge. We investigated the changes in the levels of low abundance rat brain cytosolic proteins after administration of kainic acid, a potent neurotoxin and excitatory amino acid. The cytosolic proteins from controls and animals treated with kainic acid were fractionated on an ion-exchange column. The fractions collected were analyzed by 2D electrophoresis, and the proteins with altered levels were identified by matrix-assisted laser desorption ionization or ion-spray MS. We found a manifold decrease in annexin VII, heat shock cofactor HOP/p60 and SP-22 and a manifold increase in heparin-binding protein p30. The results suggest, respectively, the involvement of an apoptotic pathway, recruitment of the heat-shock protein machinery, generation of an antioxidant response, and, probably, induction of repair mechanisms. Three of the four proteins with altered levels had not been previously detected in the cytosolic fraction, and detection of the altered levels was possible only after the protein-enriching step. PMID- 11422384 TI - In vitro processing of the proproteins GrdE of protein B of glycine reductase and PrdA of D-proline reductase from Clostridium sticklandii: formation of a pyruvoyl group from a cysteine residue. AB - GrdE and PrdA of Clostridium sticklandii are subunits of glycine reductase and D proline reductase, respectively, that are processed post-translationally to form a catalytic active pyruvoyl group. The cleavage occurred on the N-terminal side of a cysteine residue, which is thus the precursor of a pyruvoyl moiety. Both proproteins could be over-expressed in Escherichia coli and conditions were developed for in vitro processing. GrdE could be expressed as full-size protein, whereas PrdA had to be truncated N-terminally to achieve successful over expression. Both proproteins were cleaved at the in vivo observed cleavage site after addition of 200 mM NaBH4 in Tris buffer (pH 7.6) at room temperature as analysed by SDS/PAGE and MS. Cleavage of GrdE was observed with a half-time of approximately 30 min. Cys242, as the precursor of the pyruvoyl group in GrdE, was changed to alanine, serine, or threonine by site-directed mutagenesis. The Cys242 ->Ser and Cys242-->Thr mutant proteins were also cleaved under similar conditions with extended half-times. However, the Cys242-->Ala mutant protein was not cleaved indicating a pivotal role of the thiol group of cysteine or hydroxyl group of serine and threonine during the processing of pyruvoyl group-dependent reductases. PMID- 11422385 TI - Mechanism of fatty acid desaturation in the green alga Chlorella vulgaris. AB - The hypothesis that the Delta9 desaturase of Chlorella vulgaris might operate by a synchronous mechanism has been tested using a kinetic isotope effect (KIE) approach. Thus the intermolecular primary deuterium KIE on the individual C-H bond cleavage steps involved in Delta9 desaturation have been determined by incubating growing cultures of C. vulgaris (strain 211/8K) with mixtures of the appropriate regiospecifically deuterated fatty acid analogues. Our analysis shows that the introduction of a double bond between C-9 and C-10 occurs in two discrete steps as the cleavage of the C9-H bond is very sensitive to isotopic substitution (kH/kD = 6.6 +/- 0.3) whereas a negligible isotope effect (kH/kD = 1.05 +/- 0.05) was observed for the C10-H bond-breaking step. Similar results were obtained for linoleic acid biosynthesis (Delta12 desaturation). These data clearly rule out a synchronous mechanism for these reactions. PMID- 11422386 TI - Cleavage of AML1/MTG8 by asymmetric hammerhead ribozymes. AB - The chromosomal translocation t(8;21) is one of the most frequent aberrations associated with acute myeloid leukaemia. It joins the 5' section of the AML1 gene with the almost complete open reading frame of MTG8 (ETO). The resulting fusion RNA represents a leukaemia-specific target for antisense/ribozyme inhibition. We tested several asymmetric hammerhead ribozymes targeted against the fusion site for their ability to cleave the AML1/MTG8 RNA at low magnesium concentrations. One ribozyme cleaves AML1/MTG8 RNA with high catalytic efficiency without binding or cleaving the wild-type AML1 transcript. The presence of cellular RNA does not affect the cleavage. Injection of AML1/MTG8 RNA and ribozyme RNA into Xenopus eggs or oocytes causes a specific reduction of AML1/MTG8 protein expression. Asymmetric anti-AML1/MTG8 ribozymes may be valuable modulators of AML1/MTG8 expression in leukaemic cells. PMID- 11422387 TI - Site-directed mutagenesis of the ionizable groups in the active site of Zymomonas mobilis pyruvate decarboxylase: effect on activity and pH dependence. AB - Pyruvate decarboxylase (PDC, EC 4.1.1.1) is a thiamin diphosphate-dependent enzyme about which there is a large body of structural and functional information. The active site contains several absolutely conserved ionizable groups and all of these appear to be important, as judged by the fact that mutation diminishes or abolishes catalytic activity. Previously we have shown [Schenk, G., Leeper, F.J., England, R., Nixon, P.F. & Duggleby, R.G. (1997) Eur. J. Biochem. 248, 63-71] that the activity is pH-dependent due to changes in kcat/Km while kcat itself is unaffected by pH. The effect on kcat/Km is determined by a group with a pKa of 6.45; the identity of this group has not been determined, although H113 is a possible candidate. Here we mutate five crucial residues in the active site with ionizable side-chains (D27, E50, H113, H114 and E473) in turn, to residues that are nonionizable or should have a substantially altered pKa. Each protein was purified and characterized kinetically. Unexpectedly, the pH-dependence of kcat/Km is largely unaffected in all mutants, ruling out the possibility that any of these five residues is responsible for the observed pKa of 6.45. We conjecture that the kcat/Km profile reflects the protonation of an alcoholate anion intermediate of the catalytic cycle. PMID- 11422388 TI - Structural analysis of the carbohydrate components of the outer membrane of the lipopolysaccharide-lacking cellulolytic ruminal bacterium Fibrobacter succinogenes S85. AB - The polysaccharides from the outer membrane of the Gram-negative ruminal bacterium Fibrobacter succinogenes were isolated by phenol/water extraction and separated by size-exclusion chromatography in the presence of deoxycholate detergent into a lower-molecular-mass fraction designated 'glycolipid' and a high molecular-mass 'capsular polysaccharide' fraction. Both fractions lacked typical lipopolysaccharide components including 2-keto-3-deoxyoctulosonic acid and 3 hydroxy fatty acids. Carbohydrate components of these fractions were represented by two polysaccharides and one oligosaccharide (possibly glycolipid) with the following structures: : : where HEAEP is N-(2-hydroxyethyl)-2 aminoethylphosphonic acid, found for the first time in natural compounds. The polysaccharides contained pentadecanoic acid and anteisopentadecanoic acid, possibly present as the acyl components. All constituent monosaccharides except L rhamnose had a D-configuration. In addition to having a structural role in the outer membrane, these polysaccharides may provide protection for this lipopolysaccharide-less bacterium in the highly competitive ruminal environment, as phosphonic acids covalently linked to membrane polymers have in the past been attributed the function of stabilizing membranes in the presence of phosphatases and lipases. PMID- 11422389 TI - 2-Methylisocitrate lyases from the bacterium Escherichia coli and the filamentous fungus Aspergillus nidulans: characterization and comparison of both enzymes. AB - In Escherichia coli and Aspergillus nidulans, propionate is oxidized to pyruvate via the methylcitrate cycle. The last step of this cycle, the cleavage of 2 methylisocitrate to succinate and pyruvate is catalysed by 2-methylisocitrate lyase. The enzymes from both organisms were assayed with chemically synthesized threo-2-methylisocitrate; the erythro-diastereomer was not active. 2 Methylisocitrate lyase from E. coli corresponds to the PrpB protein of the prp operon involved in propionate oxidation. The purified enzyme has a molecular mass of approximately 32 kDa per subunit, which is lower than those of isocitrate lyases from bacterial sources ( approximately 48 kDa). 2-Methylisocitrate lyase from A. nidulans shows an apparent molecular mass of 66 kDa per subunit, almost equal to that of isocitrate lyase of the same organism. Both 2-methylisocitrate lyases have a native homotetrameric structure as identified by size-exclusion chromatography. The enzymes show no measurable activity with isocitrate. Starting from 250 mM pyruvate, 150 mM succinate and 10 microM PrpB, the enzymatically active stereoisomer could be synthesized in 1% yield. As revealed by chiral HPLC, the product consisted of a single enantiomer. This isomer is cleaved by 2 methylisocitrate lyases from A. nidulans and E. coli. The PrpB protein reacted with stoichiometric amounts of 3-bromopyruvate whereby the activity was lost and one amino-acid residue per subunit became modified, most likely a cysteine as shown for isocitrate lyase of E. coli. PrpB exhibits 34% sequence identity with carboxyphosphoenolpyruvate phosphonomutase from Streptomyces hygroscopicus, in which the essential cysteine residue is conserved. PMID- 11422390 TI - Molecular cloning of bovine zona pellucida glycoproteins ZPA and ZPB and analysis for sperm-binding component of the zona. AB - The zona pellucida, a transparent envelope surrounding the mammalian oocyte, comprises three glycoproteins, ZPA, ZPB and ZPC, and plays important roles in fertilization. We have previously reported that apparent relative molecular masses of bovine zona glycoproteins on SDS/PAGE under nonreducing conditions after removal of poly N-acetyllactosamine at the nonreducing portion of sugar chains with endo-beta-galactosidase are 72 000, 58 000 and 45 000 [Noguchi, S., Yonezawa, N., Katsumata, T., Hashizume, K.,Kuwayama, M., Hamano, S., Watanabe, S. & Nakano, M. (1994) Biochim. Biophys. Acta 1201, 7-14]. The N-terminal amino-acid sequences and crossreactivity to antibodies specific to each porcine zona component show that the bovine components correspond to porcine ZPA, ZPB and ZPC, respectively. In this study, we deduced amino-acid sequences of bovine ZPA and ZPB by cDNA cloning and sequencing. Identities in amino-acid sequences between bovine and porcine counterparts were 77% for ZPA and 75% for ZPB, whereas between bovine and murine counterparts identities were 57% for ZPA and 37% for ZPB. The positions of Cys were completely conserved in bovine ZPA and ZPB compared with counterparts of other mammalian species. Bovine ZPA was processed between Ala and Asp on fertilization, suggesting that the consensus motif for the processing is Ala-Asp-Asp/Glu. We purified bovine zona components and examined their sperm binding activity with an in vitro competition assay and sperm-bead-binding assay. As a result, ZPB showed the strongest sperm-binding activity among the components. ZPC also showed sperm-binding activity and the activity per molecule was about one-sixth that of ZPB according to the result of the sperm-bead-binding assay. We could not determine if ZPA has significant sperm-binding activity, but the activity may be much lower than that of ZPB even if ZPA has significant activity. Thus, ZPB may play a major role in sperm binding in bovine zona. PMID- 11422391 TI - Crystal structures of recombinant and native soybean beta-conglycinin beta homotrimers. AB - The crystal structures of recombinant and native beta homotrimers of soybean beta conglycinin were determined by X-ray crystallography at 2.7 and 2.8 A resolutions, respectively. The crystals of the recombinant and native beta homotrimers belong to space group P21 with cell parameters a = 80.51 A, b = 63.48 A, c = 131.43 A, and beta = 90.01 degrees and with cell parameters a = 82.78 A, b = 69.47 A, c = 125.33 A and beta = 97.22 degrees, respectively. The beta monomers consist of amino-terminal and carboxyl-terminal modules that are very similar to each other and are related by a pseudo-dyad axis. Each module of the beta monomer is subdivided into a core and a loop domain. These structural features of both beta homotrimers are consistent with those of canavalin and phaseolin, which are similar vicilin class proteins. The superposition of the models of the native and recombinant beta monomers shows a root mean square deviation of 0.43-0.51 A for 343 common Calpha atoms within 2.0 A. This result indicates that the N-linked glycans do not influence the final structure of the beta homotrimer. Comparison of the models of beta-conglycinin, phaseolin and canavalin indicates that beta conglycinin resembles canavalin rather than phaseolin, and that canavalin and phaseolin differ the most among them. The evolutional relationships are discussed. PMID- 11422392 TI - Poly(A) polymerase from Escherichia coli adenylylates the 3'-hydroxyl residue of nucleosides, nucleoside 5'-phosphates and nucleoside(5')oligophospho(5')nucleosides (NpnN). AB - The capacity of Escherichia coli poly(A) polymerase to adenylylate the 3'-OH residue of a variety of nucleosides, nucleoside 5'-phosphates and dinucleotides of the type nucleoside(5')oligophospho(5')nucleoside is described here for the first time. Using micromolar concentrations of [alpha-32P]ATP, the following nucleosides/nucleotides were found to be substrates of the reaction: guanosine, AMP, CMP, GMP, IMP, GDP, CTP, dGTP, GTP, XTP, adenosine(5')diphospho(5')adenosine (Ap2A), adenosine (5')triphospho(5')adenosine (Ap3A), adenosine(5')tetraphospho(5')adenosine (Ap4A), adenosine(5')pentaphospho(5')adenosine (Ap5A), guanosine(5')diphospho(5') guanosine (Gp2G), guanosine(5')triphospho(5')guanosine (Gp3G), guanosine(5')tetraphospho(5')guanosine (Gp4G), and guanosine(5')pentaphospho(5')guanosine (Gp5G). The synthesized products were analysed by TLC or HPLC and characterized by their UV spectra, and by treatment with alkaline phosphatase and snake venom phosphodiesterase. The presence of 1 mM GMP inhibited competitively the polyadenylylation of tRNA. We hypothesize that the type of methods used to measure polyadenylation of RNA is the reason why this novel property of E. coli poly(A) polymerase has not been observed previously. PMID- 11422393 TI - Differentiation between conformational and autoproteolytic stability of the neutral protease from Bacillus stearothermophilus containing an engineered disulfide bond. AB - The introduction of a disulfide bond into the neutral protease from Bacillus stearothermophilus by the double mutation G8C/N60C had resulted in an extremely thermostable enzyme with a half-life of 35.9 min at 92.5 degrees C [Mansfeld, J., Vriend, G., Dijkstra, B.W., Veltman, O.R., van den Burg, B., Venema, G., Ulbrich Hofmann, R. & Eijsink, V.G. (1997) J. Biol. Chem. 272, 11152-11156]. The study in guanidine hydrochloride of this enzyme and the respective wild-type enzyme allowed us to distinguish between the stability toward global unfolding and autoproteolysis. At low protease concentrations (20 microg.mL-1) and short periods of incubation with guanidine hydrochloride (5 min), transition curves without the interference by autoproteolysis could be derived from fluorescence emission measurements. The effect of the disulfide bond on the global unfolding of the protein proved to be smaller than expected. In contrast, the measurement of autoproteolysis at higher protein concentrations (100 microg.mL-1) by quantitative evaluation of the bands of intact protein on SDS/PAGE revealed a strong stabilization toward autoproteolytic degradation by the disulfide bond. The rate of autoproteolysis in guanidine hydrochloride was found to be much lower than that of thermal denaturation, which can be attributed to the inhibition of the proteases by this denaturant. The results suggest that the disulfide bond stabilizes the protease against autoproteolysis more than against global unfolding. Autoproteolysis starts as soon as the cleavage sites in flexible external structural regions become accessible. It is suggested that the stabilizing effect of the disulfide bond is caused by the fixation of the crucial loop region 56-69 or by hindrance of the primary cleavage in this region by the amino acid exchanges. PMID- 11422394 TI - Plaque cholesterol and calcium: the value of EBCT in the detection of coronary atherosclerosis. PMID- 11422395 TI - Electron-beam computed tomography of the heart: what do we see and what is concealed? PMID- 11422396 TI - Quantification of coronary artery calcification in patients with FH using EBCT. AB - BACKGROUND: Calcification of the coronary vessel wall is regarded as a marker of advanced coronary atherosclerosis. METHODS: To test whether patients with heterozygous familial hypercholesterolemia (FH) exhibit excessive calcification of the coronary vessel wall, we quantified coronary artery calcium in LDL apheresis treated FH-patients with known severe coronary artery disease (CAD) (n = 10), in patients with moderate hypercholesterolemia and known severe CAD (n = 10), and in asymptomatic controls (n = 10) using electronic beam CT. The total coronary calcium score (Agatston-Score), the number of calcified lesions and the calcified plaque volume were evaluated for this study. RESULTS: CAD-patients with FH, although on average 10 years younger, had a significantly higher total coronary calcium score (702/2018/2890), number of lesions (34/43/49) and calcified plaque volume (700/1818/2313) compared to patients with CAD only (480/641/1362, 10/16.5/22, 480/588/1209, respectively) and controls (10/47/137, 2/4/10, 15/50/144, respectively). Furthermore, we observed a significant correlation (r = 0.93; P < 0.01) between LDL-cholesterol levels (pretreatment levels of the CAD-FH group) and the total coronary calcium score in all three groups. Our results demonstrate that coronary artery calcification is more extensive in CAD-patients with FH than in CAD-patients with moderate hypercholesterolemia. In addition, we provide evidence that the amount of calcium in the coronary vessel wall in FH patients result from a long lasting history of elevated LDL-Cholesterol levels. CONCLUSION: These findings emphasize the significance of LDL-cholesterol as a risk factor for atherosclerosis and underline the importance of early diagnosis of CAD and early cholesterol lowering therapy in FH patients. PMID- 11422397 TI - Aspects of gene polymorphisms in cardiovascular disease: the renin-angiotensin system. AB - The renin-angiotensin system (RAS) plays a central role in cardiovascular homeostasis. Angiotensin is the key peptide of the RAS, and exerts its influence on the heart and blood vessels both through its haemodynamic effects (via its influence on after-load and pre-load and determining coronary vasoconstriction) and through its direct cellular effects (via its actions on cell proliferation). Numerous studies in the past 10 years have demonstrated that the pharmacological inhibition of angiotensin converting enzyme (ACE), one of the two critical enzymes of the RAS, improves the outcome in patients with several cardiovascular disorders (hypertension, heart failure, ischaemic heart disease). These studies suggest a role of the RAS as a major determinant of cardiovascular risk. Recent data suggest that genetics may in turn contribute to modulating the effects of angiotensin on coronary vascular biology and ischaemia. This paper reviews the physiologic characteristics of the RAS and recent research developments related to angiotensin cell biology and pathobiology in heart disease. In particular, this review will cover the genetic aspects of RAS and their implications in cardiovascular disease. PMID- 11422398 TI - Mitochondrial DNA deletion associated with the reduction of adenine nucleotides in human atrium and atrial fibrillation. AB - BACKGROUND: Structural changes in the number, size, and shape of mitochondria (mt) have been observed in the atrial muscles of patients with atrial fibrillation (AF) and of animals with rapid atrial pacing, however, it is not known whether the mitochondrial function is impaired in human atrium with AF. MATERIALS AND METHODS: We determined adenine nucleotides concentrations and mtDNA deletions in 26 human right atria obtained at the time of cardiac surgery, using HPLC and PCR amplification, and studied the relationship between mtDNA deletions and clinical manifestations, the haemodynamic parameters of the patients and adenine nucleotide concentrations in their atrium. RESULTS: The age and the prevalence of AF were significantly higher in the patients with a mtDNA deletion of 7.4 kb than in those without a deletion; there were no significant differences regarding haemodynamic parameters between the two groups. The concentrations of ATP, ADP, AMP and total adenine nucleotides in the right atrium were significantly lower in the patients with mtDNA deletions than the patients without a deletion. In a gender- and diseased-matched population, the mtDNA deletion was still significantly associated with age and a decreased concentration of adenine nucleotides in the atrium. Using quantitative PCR analysis, the proportion of mtDNA deletion to normal mtDNA of the atrium, was estimated to be 0.3-2% in four cases. CONCLUSION: These results suggest that the deletion of mtDNA associated with ageing or AF can lead to a bioenergetic deficiency due to an impaired ATP synthesis in the human atrium; however, no conclusion can be made whether mtDNA deletion were the result or the cause of an impaired ATP synthesis, ageing, hemodynamic deterioration, or AF. PMID- 11422399 TI - Leucine suppresses acid-induced protein wasting in L6 rat muscle cells. AB - BACKGROUND: Metabolic acidosis induces protein wasting in skeletal muscle cells, accompanied by decreased glycolysis and compensatory increased consumption of other metabolic fuels, implying that protein wasting arises from fuel starvation and might be rectified by fuel supplements. Design To test this hypothesis, total protein and protein degradation (release of 14C-phenylalanine) were measured in L6 skeletal muscle cells cultured in Eagle's Minimum Essential Medium at pH 7.1 7.5 for 3 days with metabolic inhibitors or metabolic fuel supplements. RESULTS: Inducing metabolic fuel starvation with inhibitors (1 mmol L(-1) 2-deoxyglucose or 0.1 mmol L(-1) KCN [potassium cyanide]) failed to stimulate protein degradation or net protein wasting under nonacidaemic conditions (pH 7.5). Conversely metabolic fuel supplements (1 mmol L(-1) octanoate, pyruvate or alanine) failed to increase the protein content of the cultures at any pH tested, in spite of significant consumption of the fuels by the cells. Only leucine (1-3 mmol L(-1)) increased protein content and suppressed protein degradation in opposition to the catabolic effect of acidaemia (pH 7.1). Conclusion Leucine exerts a beneficial anabolic effect on cultured skeletal muscle cells in the face of metabolic acidaemia. The failure of other metabolic fuels to do this, and of the metabolic inhibitors to exert a catabolic effect, suggests that leucine acts as a specific modulator of protein turnover and not as a nonspecific source of carbon for oxidation as a fuel. PMID- 11422400 TI - Role of various tissues in apo(a) fragmentation and excretion of fragments by the kidney. AB - BACKGROUND: Lipoprotein(a) [Lp(a)] is an atherothrombotic plasma lipoprotein with unknown function. Little is known about the catabolism of this lipoprotein, in particular the steps related to apolipoprotein(a) [apo(a)] fragmentation and excretion by the kidney. MATERIAL AND METHODS: High plasma levels (up to 9 mg dL( 1)) of the N-terminal fragment of apo(a) were expressed in mice by adenovirus mediated gene transfer. Plasma of such N-apo(a) mice was injected into acceptor mice and the fragmentation and urinary secretion of N-apo(a) were followed by immunochemical techniques. RESULTS: Mice transduced with N-Ad expressed apo(a) fragments with 3-11 kringle-IV (KIV) repeats. Injection of N-apo(a)-plasma from donor mice into acceptor mice resulted in fragmentation of N-apo(a)s with 3-11 KIVs yielding smaller peptides down to 2 KIVs. Secretion of N-apo(a)-fragments with 2 to maximally 6 KIVs into urine occurred as early as 2 min after injection. Immunohistochemical studies of kidney suggested filtration as a mechanism of apo(a)-fragment excretion. When N-apo(a) was incubated in vitro with various tissues from perfused mice, skeletal muscle and kidney followed by liver and spleen contributed to fragmentation. Tissues from unperfused organs, or the addition of normal mouse plasma, caused marked reduction in N-apo(a) fragmentation. EDTA, and not aprotinin or leupeptin, prevented apo(a) cleavage. CONCLUSION: Here we provide evidence that apo(a) is cleaved by metalloproteinases located on skeletal muscle, kidney and other organs. Small apo(a)-fragments up to a size of 6 KIVs are excreted into urine, yet a major portion of apo(a) fragments is removed from circulation extrarenally. PMID- 11422401 TI - Effects of OPB-9195, anti-glycation agent, on experimental diabetic neuropathy. AB - BACKGROUND: Nonenzymatic glycation of neural proteins and their end-products (advanced glycation end-products, AGE) have been implicated in the pathogenesis of diabetic neuropathy. We need a development of effective ant-glycation agents for future clinical use. MATERIALS AND METHODS: We examined the effects of OPB 9195 (OPB), a new inhibitor of glycation, on the peripheral nerve structure and function in diabetic rats. Eight-week-old Wistar rats were made diabetic by streptozotocin (40 mg kg(-1), i.v.) and OPB (60 mg kg(-1) day(-1)) was given by gavage for 24 weeks. Age- and sex-matched normal Wistar rats were used for comparison. RESULTS: During the experimental period, OPB treatment did not affect the reduced body weight, elevated levels of blood glucose and glycated haemoglobin in diabetic rats. At the end of the experiment, delayed tibial motor nerve conduction velocity was significantly improved (by 60%) in treated diabetic rats, with reduction of serum AGE levels. Expression of immunoreactive AGE in the sciatic nerve was reduced in treated diabetic rats compared with those in untreated rats. Sciatic nerve (Na+, K+)-ATPase activity was also restored in treated diabetic rats. On the cross-sectioned sciatic nerves, positive cells with oxidative stress-related DNA damage, as expressed by 8-hydroxy-2'-deoxyguanosine, were less in the peripheral nerve of treated diabetic rats compared with those of untreated rats. CONCLUSION: The current study suggested that OPB is beneficial for the reduction of serum AGE and the prevention of diabetic neuropathy. PMID- 11422402 TI - Fatty acid binding protein 2 and insulin resistance. AB - BACKGROUND: An A54T polymorphism of the fatty acid binding protein 2 (FABP2) gene was found to be associated with insulin resistance in nondiabetic Pima Indians. Design This is a cross-sectional study to examine the role of this polymorphism in insulin resistance in 71 healthy and normotensive Caucasian subjects with normal glucose tolerance. Insulin sensitivity (%S, ISI(M), ISI(S)) and beta-cell function (%B, dI/dG, 1stPHS, 2ndPHS) were estimated based on published models. Their genotypes were determined using a polymerase chain reaction-restriction fragment length polymorphism assay. The relationship between genotypes and phenotypes was examined. RESULTS: After genotyping, we identified 34 AA, 32 AT and five TT subjects. The TT subjects were pooled together with the AT subjects during the analysis due to their low number. No difference was noted in gender distribution, clinical features, or fasting lipid profile between the two genotypic groups (AA vs. AT/TT). The AT/TT group had lower %S and ISI(S) than the AA group (P = 0.0118 and P = 0.0170, respectively). The difference in ISI(M) was marginal (P = 0.0544). However, no difference was noted in beta-cell function between the two groups. Multivariate analysis revealed that this polymorphism was an independent but modest determinant for %S (P = 0.0149), ISI(M) (P = 0.0489) and ISI(S) (P = 0.0175). It independently contributed 6.04% (95% CI, 0.02 20.53%), 4.28% (95% CI, 0.08-17.63%) and 4.94% (95% CI, 0.01-18.75%) of the variation of %S, ISI(M) and ISI(S), respectively. CONCLUSIONS: We demonstrated that the A54T polymorphism at the FABP2 locus is a risk factor for insulin resistance in a Caucasian population. PMID- 11422403 TI - Noninvasive diagnosis of the degree of hepatic fibrosis using ultrasonography in patients with chronic liver disease due to hepatitis C virus. AB - BACKGROUND: Chronic liver disease is characterized by progressive hepatic fibrosis and changes in hepatic haemodynamics. This study has addressed the possibility of a noninvasive diagnosis of the degree of hepatic fibrosis by evaluating the velocity of blood in the hepatic vasculature. Materials and methods The maximum velocity of blood at the portal vein and hepatic artery was measured in 80 patients with chronic liver diseases (19 with liver cirrhosis; 61 with chronic hepatitis) and in 20 normal volunteers by Doppler ultrasonography. The arterio-portal ratio (A/P ratio) was calculated by dividing the maximum velocity of blood (Vmax) in the hepatic artery with the Vmax in the portal vein. Multivariate analysis was used to disclose the independent predictors of the degree of hepatic fibrosis. RESULTS: The levels of A/P ratio were significantly higher in patients with liver cirrhosis (LC) compared to those with chronic hepatitis (CH) and normal controls. Probit analysis revealed that the value of A/P ratio at which CH becomes LC was A/P >or= 3.5. The levels of A/P ratio were also significantly higher in patients with severe fibrosis compared with mild (P < 0.0001) and moderate (P < 0.0001) fibrosis. Multivariate analysis disclosed right A/P ratio (P = 0.0001), left A/P ratio (P = 0.013), and platelet counts (P = 0.0172), as the only independent predictors of the degree of hepatic fibrosis. CONCLUSIONS: A/P ratio may be used for the noninvasive diagnosis of the degree of hepatic fibrosis in patients with chronic liver diseases. PMID- 11422404 TI - Interferon-gamma in healthy subjects: selective modulation of inflammatory mediators. AB - BACKGROUND: It is suggested that interferon-gamma (IFN-gamma), like other cytokines, is a mediator in the host inflammatory response, which could be of importance in the pathophysiology of sepsis. The role of IFN-gamma in human host inflammatory responses, however, has not been studied. DESIGN: In a placebo controlled trial we studied the acute effects of IFN-gamma administration on host inflammatory mediators in healthy men: i.e. the cytokine/chemokine cascade system, acute-phase proteins, activation markers of the innate cellular immunity and coagulation/fibrinolysis parameters. RESULTS: IFN-gamma increased plasma levels of interleukin-6 (IL-6), IL-8 and IFN-gamma-inducible protein-10 (IP-10) (P < 0.05), but did not affect plasma levels of other cytokines (IL-4, IL-10, tumour necrosis factor-alpha, IL-12p40/p70). Plasma concentrations of C-reactive protein and secretory phospholipase A2 both increased (P < 0.05). Plasma levels of the leucocyte activation marker elastase-alpha1-antitrypsin complexes increased after IFN-gamma administration (P < 0.05), IFN-gamma increased the percentage of high-affinity Fcgamma-receptor (FcgammaRI) -positive neutrophils (P < 0.05), but did not affect the mean fluorescence intensity of FcgammaRI on neutrophils. Procoagulant and profibrinolytic effects of IFN-gamma were evidenced by increased plasma levels of prothrombin fragment F1 + F2, tissue-plasminogen activator and plasmin-alpha2-antiplasmin complexes (P < 0.05). CONCLUSION: We conclude that IFN-gamma selectively affects host inflammatory mediators in humans. PMID- 11422405 TI - Impaired nutritional status in common variable immunodeficiency patients correlates with reduced levels of serum IgA and of circulating CD4+ T lymphocytes. AB - BACKGROUND: Common variable immunodeficiency (CVI) is a primary defect of the immune system. Infections, persistent diarrhoea and malabsorption may result in malnutrition, which may in turn contribute to increased morbidity. In this paper, the prevalence of malnutrition in CVI was evaluated. PATIENTS AND METHODS: Forty CVI patients (20 male, 20 female, aged 17-75 years) underwent anthropometric measurements from which body mass index, arm fat and muscle area were calculated. Body mass index values < 18.5 and arm fat and muscle area values < 10th percentile were considered indicative of malnutrition. Patients were divided into four groups according to circulating CD4+ T cells (lower or greater than 300 microL(-1)) and serum immunoglobulin A (IgA) levels (detectable and undetectable). RESULTS: Body mass index < 18.5, arm fat and muscle area < 10th percentile were observed in 23%, 58% and 44%, respectively, of patients. Lower values of body mass index, arm fat and muscle area were more frequent in patients with low CD4+ cells and undetectable IgA. Low arm fat values were more frequent in patients with diarrhoea (P = 0.03). Infectious episodes were more frequent in undetectable IgA than in detectable IgA patients (P = 0.04). CONCLUSIONS: Anthropometric measurements revealed an increased rate of malnutrition in CVI patients, particularly in those with low CD4+ and undetectable IgA, suggesting that selected CVI subjects could be considered for standard or specialized nutritional support. PMID- 11422406 TI - Effects of smoking on activation markers, Fas expression and apoptosis of peripheral blood lymphocytes. AB - BACKGROUND: Smoking influences numbers and function of peripheral blood lymphocytes (PBL) by a process that is badly understood. We conducted this study to evaluate whether the immune impairment of smoking might be related to changes in the expression or functionality of Fas, a cell surface molecule that plays a central role in immune homeostasis and cytotoxic activity. METHODS: PBL from 10 smoking and 10 nonsmoking healthy volunteers were isolated. Flow cytometry was performed to measure the state of activation, Fas expression and apoptosis of PBL. Functionality of Fas was tested by assessing apoptosis after incubation of isolated lymphocytes with agonistic anti-Fas antibodies in four smoking and four nonsmoking individuals. RESULTS: Smoking was associated with an increase in the percentage of Fas-expressing CD4+ T and B lymphocytes. A decrease in the percentage of activated (CD38+) B cells was observed. In vitro Fas-induced apoptosis did not appear different between smokers and nonsmokers. No differences in the percentages of circulating apoptotic lymphocytes could be demonstrated between smoking and nonsmoking individuals. Conclusion Smoking is associated with increased Fas expression on PBL in general, and on B cells in particular. This might render these cells more susceptible for apoptosis. As Fas is functionally intact this may also explain the reduced percentage of activated (CD38+) B cells found in smoking individuals. The latter may contribute to the reduced humoral immune response observed in smokers. PMID- 11422407 TI - Immunomodulatory effects of moxifloxacin in comparison to ciprofloxacin and G-CSF in a murine model of cyclophosphamide-induced leukopenia. AB - We analyzed the effect of the two quinolones moxifloxacin and ciprofloxacin on the repopulation of hematopoietic organs and on the production of cytokines by various organs of cyclophosphamide (CP)-induced leukopenic mice. The effect was compared to that of G-CSF. Cyclophosphamide injection induced a severe leukopenia, with nadir at day 4 post-injection. All the quinolone and G-CSF treated animals showed WBC>500/microL at the nadir, compared to 50% of saline treated mice. Cyclophosphamide induced a marked decrease in the number of myeloid progenitors (CFU-C) in bone marrow (BM) and spleen. Quinolone or G-CSF treatment resulted in a 1.4-4.3-fold increase in CFU-C numbers in the BM; no enhancement was observed in the spleen. Treatment with CP resulted in enhanced colony stimulating activity (CSA) in bone shaft and spleen and decreased activity in bladder and lung. Treatment of CP-injected mice with quinolones significantly enhanced CSA in the bone shaft, spleen, lung and bladder on different days. In normal mice the highest levels of GM-CSF and IL-6 were observed in lung conditioned medium (compared to bone shaft, spleen and bladder). Injection of CP resulted in a 22.5- and 93-fold decrease in GM-CSF and IL-6 levels, respectively, in lung-conditioned medium, while treatment with quinolones resulted in 2-4-fold increase in GM-CSF with no effect on IL-6 production. G-CSF treatment had no enhancing effect on GM-CSF nor on IL-6 production. We conclude that moxifloxacin and ciprofloxacin administered to CP-injected mice revert some of the immune suppressive effects of cyclophosphamide. PMID- 11422408 TI - Identification of numerical and structural chromosome aberrations in 15 high hyperdiploid childhood acute lymphoblastic leukemias using spectral karyotyping. AB - Spectral karyotyping (SKY) on metaphase spreads from 15 high hyperdiploid (>51 chromosomes) childhood acute lymphoblastic leukemias (ALL), which typically display a poor chromosome morphology, was performed in order to investigate the pattern of numerical abnormalities, reveal the chromosomal origin of marker chromosomes, and identify translocations and other interchromosomal rearrangements not detected by G-banding analysis. In all cases the numerical changes could be fully characterized, and a non-random pattern of chromosomal gain was identified, with chromosomes X, 21, 14, 17, 6, 18, 4, and 10 being most frequently gained. The numerical changes had been partly misinterpreted in 12 of the 15 ALL patients using G-banding, and the present study hence emphasizes the importance of SKY in identifying such anomalies, some of which, i.e. +4 and +10, have been suggested to be prognostically important. The chromosomal origin of all marker chromosomes and of seven structural rearrangements, one of which was the prognostically important Philadelphia chromosome, could be identified. Five rearrangements [der(1)t(1;14)(q32;q21), der(2)t(2;8)(q36;?), der(3)t(2;3)(q21;?), der(8)t(8;14)(?;?), and t(9;21)(q12;q22)] have previously not been reported in ALL, emphasizing the value of SKY in identifying novel chromosomal rearrangements. PMID- 11422409 TI - Haemorheological adaptation during pregnancy in a Latin American population. AB - OBJECTIVE: To investigate haemorheological changes during pregnancy in a Latin American population and compare to previously published data from Caucasian populations. DESIGN: Cross-sectional study. POPULATION: 75 pregnant women at 10 36 wk of gestation and 17 non-pregnant female controls in Lima, Peru. All the women and their ancestors for three generations were born and lived at sea level. METHODS: Viscosity, haematocrit and plasma fibrinogen, albumin and total protein concentrations were determined in blood samples obtained after an overnight period of fasting. RESULTS: At 10 wk of gestation, total protein concentration and plasma viscosity were above non-pregnant levels by about 15% and subsequently decreased linearly with gestation. Fibrinogen concentration was increased in the first trimester; it then decreased to a nadir at about 20 wk and subsequently increased. Albumin concentration decreased linearly with gestation. Haematocrit decreased from pre-pregnancy levels at 10 wk to a nadir at about 26 wk. Blood viscosity increased in the first trimester and then decreased with gestation to a nadir at about 26 wk. CONCLUSION: In the first trimester of pregnancy blood and plasma viscosity are increased and they subsequently fall with advancing gestation. Plasma viscosity reflects the changes in total protein concentration, and blood viscosity is dependent on the interplay of changes in plasma viscosity and haematocrit. PMID- 11422410 TI - Atypical hemoglobin H disease in a Thai patient resulting from a combination of alpha-thalassemia 1 and hemoglobin Constant Spring with hemoglobin J Bangkok heterozygosity. AB - A case of hemoglobin H disease in combination with hemoglobin Constant Spring and a beta-globin chain variant is reported in a 3-yr-old Thai girl. On routine cellulose acetate electrophoresis, one abnormal band in addition to the hemoglobins A, A2, H, Bart's and Constant Spring was detected. The amount of this abnormal band movement towards more anodic to the hemoglobin A was 35.7%. DNA analysis of the alpha-globin gene cluster by polymerase chain reaction (PCR) revealed a combination of defects caused by the SEA-type alpha-thalassemia 1 and the alpha-Constant Spring gene. Analysis of beta-globin gene by PCR and DNA sequencing also detected the heterozygosity for the GGC-GAC mutation at codon 56, leading to a substitution of aspartic acid for glycine resulting in the hemoglobin J Bangkok. The hematologic data of this unusual case of hemoglobin H disease are presented and compared with two compound heterozygotes for hemoglobin J Bangkok and alpha-thalassemia 1 found in the patient's father and grandfather. A simple DNA assay based on an allele-specific PCR for rapid diagnosis of the hemoglobin J Bangkok is also described. PMID- 11422411 TI - Hydroxyurea and periodicity in myeloproliferative disease. AB - Three patients, one with polycythaemia vera (PV) and two with chronic myeloid leukaemia (CML), are described who had cycling of blood counts which became apparent whilst receiving hydroxyurea therapy. Significant periodicity was confirmed with the use of the Lomb periodogram. This is Fourier power spectral analysis tailored for unevenly sampled data. The patient with PV had marked oscillations of platelet counts with a periodicity of 29 d and an amplitude of (202-588)x10(9)/L. Smaller oscillations of neutrophil, monocyte and lymphocyte numbers and Hb levels occurred with a similar periodicity. Anticipatory changes in hydroxyurea dosage or the maintenance of a constant dose did not abolish periodicity, but a change in therapy to the non-cycle-specific drug anagrelide dampened and abolished the cycling. One of the patients with CML had tremendous and clear oscillations in white cell, platelet and Hb levels, with a mean periodicity of 74 d. The other had erratic counts which were confirmed to be significantly periodic (64 d), on spectral analysis. A change in therapy to busulphan in both these patients again dampened and abolished the cycling. Hydroxyurea, which is a cell-cycle-specific agent, probably exacerbates the periodicity which may be present in some patients with myeloproliferative disease. A change in therapy to non-cycle-acting compounds such as busulphan or anagrelide results in much more stable counts in such patients. PMID- 11422412 TI - Myelofibrosis with myeloid metaplasia in adult individuals 30 years old or younger: presenting features, evolution and survival. AB - Myelofibrosis with myeloid metaplasia (MMM) usually affects older people and is associated with a median survival of 3.5-5 yr. Survival of MMM patients 55 yr old or younger is longer, but there is no information on the youngest subset. The presenting features, evolution and survival were analyzed in 9 patients with MMM aged 30 yr or less, representing 2.8% of two series including 323 cases. Age ranged from 17 to 30 yr; 3 patients were males and 6 females. Five patients were asymptomatic and none had constitutional symptoms. Anemia was observed in 4 patients, being severe in only one. The WBC count was normal in 7 patients and 2 had mild leukocytosis; moderate thrombocytosis was observed in 7 patients. No patient showed blood blast cells or bone marrow cytogenetic abnormalities. With a median follow-up of 6.8 (range 0.8-28) yr, 2 patients died 10.7 and 9.9 yr from diagnosis, one from complications of bone marrow transplantation performed after progression of the disease and the other from gastrointestinal bleeding secondary to portal hypertension. Among remaining patients, one developed Budd-Chiari syndrome, another was lost to follow-up at 3 yr, and 5 remain asymptomatic and currently without treatment (one had been splenectomized at diagnosis). These results indicate that in most young adults with MMM the disease presents without adverse prognostic factors and may remain stable for years, which may be of interest when considering the treatment of such patients. PMID- 11422413 TI - Cost-utility analysis of high-dose melphalan with autologous blood stem cell support vs. melphalan plus prednisone in patients younger than 60 years with multiple myeloma. AB - We evaluated the costs and the cost utility of high-dose melphalan and autologous stem cell support followed by interferon maintenance relative to conventional treatment with melphalan and prednisone, in patients less than 60 yr of age with multiple myeloma. From March 1994 to July 1997, 274 patients with newly diagnosed, symptomatic multiple myeloma were enrolled in a prospective, non randomized, population-based, multicenter study to evaluate the treatment with high-dose melphalan and autologous blood stem cell support. Health-related quality-of-life was measured prior to treatment and during follow-up, using the EORTC QLQ-C30 questionnaire. Resource consumption was also recorded prospectively. The intensive treatment yielded a significant increase in median survival time from 44 to 62 months compared to conventionally treated patients. The corresponding gain in quality-adjusted life years (QALY) was found to be 1.2. Cost per QALY gained by the treatment with high-dose melphalan and autologous blood stem cell support was estimated at NOK 249,000 (USD 27,000). PMID- 11422414 TI - In multiple myeloma increased thrombopoietin (Tpo) production may be involved in the maintenance of platelet production. AB - In multiple myeloma (MM), suppression of haematopoiesis occurs as a result of expansion of malignant cells in the bone marrow. Thrombopoietin (Tpo) levels in patients with impaired platelet production are generally found to be highly elevated. To examine the circulating Tpo levels in patients with MM, Tpo levels were measured in 50 serum samples from 34 patients. Tpo levels were subsequently related to disease stage, and cell numbers and markers, i.e. platelet count, leukocyte count and haemoglobin (Hb) concentration. Elevated Tpo levels were found in association with decreased platelet counts (n=8), but also in patients with normal platelet counts (n=14). The latter group included patients without and with signs of impaired haematopoiesis, i.e. with decreased Hb concentration and decreased leukocyte count. These results show that neither platelet counts nor Tpo levels are reliable parameters to judge bone-marrow failure in patients with MM. Furthermore, in patients with MM, increased Tpo levels may play a role in the maintenance of thrombocytopoiesis. The origin of the increased Tpo levels remains to be determined. PMID- 11422415 TI - Increased serum levels of soluble Fas in progressive B-CLL. AB - Clinical progression of B-cell chronic lymphocytic leukemia (B-CLL) depends on survival and accumulation of leukemic cells, regulated in part by physical cell contact and soluble molecules. Here we have studied the Fas/FasL system in relation to clinical progression in B-CLL. Serum levels of soluble Fas (sFas) and FasL (sFasL) were determined by ELISA in 43 progressive and 40 non-progressive B CLL patients and in 21 control individuals. Correlation between sFas serum levels and clinical progression, stage and survival were statistically analyzed. We found high levels of sFas in B-CLL sera correlated with disease progression (p<0.01). In addition, higher sFas levels were found in patients in stages II, III and IV in comparison to patients in stage 0 (p<0.05, p<0.01, p<0.03, respectively). Survival was significantly shorter for patients with > or =6 ng/ml sFas serum levels, although a multivariate analysis did not show sFas to be a significant independent prognostic factor. Fresh B-CLL cells showed only low levels of membrane expression, which were not correlated to sFas levels in serum. In vitro activation of B-CLL cells increased Fas expression, as reported earlier, and induced cells to release sFas into the supernatant. In conclusion, our results indicate that sFas in serum may be a useful parameter for the prediction of clinical progression in B-CLL. PMID- 11422416 TI - Cyclosporin A in the treatment of refractory immune thrombocytopenia purpura in children. AB - Patients with refractory autoimmune thrombocytopenia do not respond to standard therapy with high-dose corticosteroids, intravenous immunoglobulin, and splenectomy. We describe the cases of two patients with refractory autoimmune thrombocytopenia treated with oral cyclosporin A (CsA) to evaluate the efficacy of this alternative therapy. Blood pressure and hepatic and renal function were in the normal range before initiation of treatment. Induction therapy with pulses of high-dose methylprednisolone was used for 3 consecutive days to improve the initial immune suppression. Gradual dose reduction of CsA, according the platelet count, minimized the long-term adverse effects of CsA. Oral CsA with pulses of high-dose methylprednisolone induced remission of the thrombocytopenia. Gradual weaning of CsA over months, according the platelet count, produced no observable adverse effects of the CsA. Rapid dose reduction caused thrombocytopenia, which resolved with higher dosages of CsA. Our cases show the efficacy of CsA for refractory immune thrombocytopenia. This therapeutic option with oral CsA as an additional salvage option may avoid splenectomy and the adverse effects of long term corticosteroids. Larger clinical investigations are necessary to establish the indications and therapeutic regimen for CsA in immune thrombocytopenia. PMID- 11422417 TI - Successful treatment of an acquired haemorrhagic diathesis due to factor X deficiency with chemotherapy. AB - A 70-yr-old woman presented with a severe haemorrhagic diathesis due to an acquired factor X deficiency. A plasma infusion study showed that exogenous factor X was eliminated very effectively from the patient's circulation. A bone marrow biopsy was consistent with plasma cell dyscrasia. Neither an abdominal fat biopsy nor the bone marrow biopsy confirmed an amyloidosis, although clinically no other diagnosis seemed possible. Treatment with intermittent chemotherapy, consisting of vincristine, cytoxan and prednisone, yielded definite clinical and laboratory improvement. PMID- 11422418 TI - Pseudomonas pickettii (Biovar VA-II): a rare cause of bacteremias in haematologic patients. PMID- 11422419 TI - Nomenclature for factors of the HLA system, 2000. PMID- 11422420 TI - Lack of association between HLA genotype and chronic fatigue syndrome. AB - Although the aetiology of chronic fatigue syndrome is controversial, evidence that infective agents including viruses may have a role in the development of the condition has led to studies seeking an association with the immunomodulatory HLA genes. In the present study, we sought to extend previous work using a well characterized patient group and modern HLA genotyping techniques. Fifty-eight patients were phenotyped for HLA A and B by microcytotoxicity and genotyped for HLA DRB, DQB and DPB by PCR oligoprobing, and the frequencies of antigens so assigned were compared with those from a control group of 134. No significant differences in HLA frequencies were found between patient and control groups. Thus, this study does not confirm previous findings of an HLA association with chronic fatigue syndrome, suggesting that neither presentation of viral antigen by HLA class I nor antigen processing genes in the HLA region is a major contributory factor in the development of the disease. PMID- 11422421 TI - Cattle MHC genes DOA and DOB: sequence polymorphisms and assignments to the class IIb region. AB - In a study of the genetic polymorphism of the second exons of the cattle DOA and DOB genes, two and four allelic variants were detected, respectively. In the predicted amino acid sequence, the DOA polymorphism corresponded to variation at the respective residue position, whereas the nucleotide substitutions in the DOB gene were non-informative. PCR-RFLP assays were developed for DOA and DOB typing, and both loci were genetically mapped to the BoLA class IIb region by linkage analysis in the International Bovine Reference Panel. The single nucleotide polymorphisms detected in the BoLA-DOA and -DOB genes enable these loci to be used as markers in genetic trait analyses. PMID- 11422422 TI - New chimpanzee MHC class I alleles cloned by polymerase chain reaction. AB - We describe polymerase chain reaction (PCR) cloning of full-length Patr-A, -B and -Cw locus alleles from a chimpanzee using one sense/antisense primer pair. Of the six alleles cloned here, two have not previously been described. Comparison of nucleotide sequences between the two novel Patr alleles and various HLA alleles suggests that the new Patr-A allele belongs to the A1/A3/A11 family, whereas the new Patr-Cw allele is not closely related to any known HLA-Cw allele. PMID- 11422423 TI - Nomenclature for factors of the HLA System, update January 2001. PMID- 11422425 TI - Linking structural, metabolic and functional changes in multiple sclerosis. AB - In patients with multiple sclerosis (MS), conventional magnetic resonance imaging (MRI) has markedly improved our ability to detect the macroscopic abnormalities of the brain and spinal cord. New quantitative magnetic resonance (MR) approaches with increased sensitivity to subtle normal-appearing white matter (NAWM) and grey matter changes and increased specificity to the heterogeneous pathological substrates of MS may give information complementary to conventional MRI. Magnetization transfer imaging (MTI) and diffusion-weighted imaging (DWI) have the potential to provide important information on the structural changes occurring within and outside T2-visible lesions. Magnetic resonance spectroscopy (MRS) adds information on the biochemical nature of such changes. Functional MRI might quantify the efficiency of brain plasticity in response to MS injury and improve our understanding of the link between structural damage and clinical manifestations. The present review summarizes how the application of these MR techniques to the study of MS is dramatically changing our understanding of how MS causes irreversible neurological deficits. PMID- 11422426 TI - EFNS Task Force on Molecular Diagnosis of Neurologic Disorders: guidelines for the molecular diagnosis of inherited neurologic diseases. First of two parts. PMID- 11422427 TI - Depression is an independent predictor of poor long-term functional outcome post stroke. AB - The influence of depression on the long-term outcome of stroke patients was examined among 390 of 486 consecutive patients aged 55-85 years. They completed, at 3 months after ischaemic stroke, a detailed medical, neurological, and radiological stroke evaluation, structured measures of emotion (Beck's Depression Inventory, BDI), handicap (Rankin scale, RS), and assessment of activities of daily living (Barthel Index, BI). Further RS and BI was evaluated at 15-month follow-up from these 390 patients and BDI in 276 patients. A group of 256 patients completed, in addition to the 15-month follow-up, a comprehensive psychiatric evaluation, including the Present State Examination 3 months after stroke. The DSM-III-R criteria were used for diagnosis of the depressive disorders. BDI identified depression (cut-off point > or = 10 for depression) in 171 (43.9%) of 390 and in 123 (44.6%) of 276 patients at 3- and 15-month follow up. DSM-III-R major depression was diagnosed in 66 (25.8%), and minor depression in 32 (12.5%), of 256 patients 3 months after stroke. Patients with BDI > or = 10, or major, but not minor, depression more often had poor functional outcome (RS > II and BI < 17) at 15 months. Poor functional outcome at 3 months also correlated with depression at 15 months. In logistic regression analysis, depression at 3 months (Beck > or = 10) correlated with poor functional outcome at 15 months (RS > II) (OR 2.5, 95% CI 1.6-3.8). More careful examination and treatment of depression in stroke patients is emphasized. PMID- 11422428 TI - Increased T cell expression of CD154 (CD40-ligand) in multiple sclerosis. AB - CD154 (CD40-ligand, gp39), expressed on activated T cells, is crucial in T cell dependent immune responses and may be involved in the pathogenesis of multiple sclerosis (MS). We studied cerebro-spinal fluid and peripheral blood T cell expression of CD154 in MS by flow cytometry. Patients with secondary progressive MS (SPMS) had constitutive CD154 expression on CD4 and CD8 T cells in blood. Constitutive CD154 expression was not observed in patients with relapsing remitting MS (RRMS) or clinically isolated syndromes (CIS) suggestive of demyelinating disease. After ex vivo activation CD154 was, however, expressed on a higher percentage of T cells from patients with CIS or RRMS than from healthy control subjects. These results suggest involvement of CD154 in the pathogenesis of MS, and the shift from a relapsing-remitting to a secondary progressive disease course may be associated with constitutive, systemic CD154 expression. PMID- 11422429 TI - Local insulin injection improves median nerve regeneration in NIDDM patients with carpal tunnel syndrome. AB - It has been suggested that insulin has an effect on nerve regeneration similar to that of nerve growth factor (NGF). Therefore, we aimed to evaluate the effectiveness of local insulin injection on median nerve in patients with non insulin-dependent diabetes (NIDDM) mellitus who have mild-to-moderate carpal tunnel syndrome (TS). We carried out a prospective, randomized, double-blind, placebo-controlled study in these patients. At the baseline, 20 mg methylprednisolone was injected directly into the carpal tunnel in all patients [n=43 (62 hands)]. A week after prednisolone, the placebo or NPH insulin (12 U) was injected into the carpal tunnel weekly for 7 weeks. The patients were followed up for 23 weeks. A significant improvement in mean median nerve motor distal latency (MNMDL), median nerve sensory velocity (MNSV), and global symptom score (GSS) occurred in both groups (with the exception of mean MNMDL in the placebo group). A more significant improvement in the mean MNMDL, MNSV, and GSS was observed in the insulin group when compared with the placebo group. This study suggests that local insulin treatment may be of great potential benefit in the improvement of nerve functions in NIDDM patients with mild-to-moderate CTS who opt for conservative treatment. PMID- 11422430 TI - Increased intracortical facilitation in patients with autosomal dominant pure spastic paraplegia linked to chromosome 2p. AB - There are at least seven clinically indistinguishable but genetically different types of autosomal dominant pure spastic paraplegia (ADPSP). In this study we investigated electrophysiological characteristics in patients with ADPSP linked to chromosome 2p (SPG4). Twelve patients from six different families with ADPSP linked to chromosome 2p and 15 control persons were included. Electromyography (EMG), motor and sensory nerve conduction, and motor evoked potentials using single and paired transcranial magnetic stimulation (PTMS) was performed. From the peripheral nervous system we found signs of motor and sensory axonal neuropathy. Motor evoked potentials disclosed greatly reduced corticospinal tract conduction velocity and amplitude of evoked potentials to the lower extremities indicating that the very marked spasticity predominantly seems to rely on dysfunction of the fast conducting axons of the pyramidal tract. PTMS showed an increased intracortical facilitation (ICF), which may reflect an impaired function of gamma-aminobutyric acid (GABA)-controlled interneuronal circuits in the motor cortex, alternatively an increased glutamatergic transmission or a compensatory recruitment of a larger number of neurones with corticospinal projections. PMID- 11422431 TI - Pulmonary function tests in Parkinson's disease. AB - Morbidity and mortality are usually caused by respiratory disorders in Parkinson's disease (PD) because of pulmonary functional impairments. The purpose of this study was to determine the effects of PD on ventilatory function and that the use of pulmonary function tests (PFT) may serve as an indicator of PD severity. PFT have been performed in 21 patients with PD (15 non-smoker and six exsmoker with 36.17 +/- 26.54 pack-years of smoking history; mean age 64.67 +/- 10.76 years) and 16 normal age-matched control subjects who never smoked. The clinical disability was indicated by a Hoehn-Yahr (H-Y) scale. MEF25% [maximal flow rate at 25% of remaining forced vital capacity (FVC)] and FEV1 (the volume of air expired during the first second of the FVC) in exsmoker PD group was lower than non-smoker PD group (P < 0.05). The two effort dependent variables' peak expiratory flow (PEF) and the maximal flow rate at 75% of the remaining FVC (MEF75%) percent predicted values were 70.66 +/- 24.15 and 69.05 +/- 24.39 in non smoker PD group whereas 90.18 +/- 17.24 and 90.00 +/- 18.97% predicted were in control group, respectively (P < 0.05). The maximal voluntary ventilation (MVV) was found to be 52.83 +/- 15.52 and 91.52 +/- 13.80% in PD and control group, respectively (P < 0.0001). MVV was the most effected parameter that was inversely correlated with the PD severity (r=-0.87, P < 0.0001). We concluded that less coordinated and less explosive muscle force has contributed to decrease in PEF and MEF75% values, and MVV decreases in PD as a result of the impaired performance and reduced efficiency during repetitive motor tasks which in part reflects abnormal agonist-antagonist muscle activity. So, spirometric studies may serve as a useful indicator of patients' neurophysiological conditions for the purpose of anticipating and preventing complications because of pulmonary impairment. PMID- 11422432 TI - A severe frontal-parietal lobe syndrome following cerebellar damage. AB - We report a case study of a frontal and parietal lobe syndrome with memory loss after unilateral left-sided cerebellar damage caused by a stroke in a patient with right cerebellar unusual developmental agenesis. The syndrome consisted of severe deficits in planning an organized sequence of events, in visuo constructive abilities and inappropriate jocularity. These changes are ascribed in part to cerebellar-pontine lesions with resulting frontal lobe diaschisis as documented by single-photon emission computed tomography in the absence of morphological damage to the neocortex. PMID- 11422433 TI - Anti-GQ1b antibody status, magnetic resonance imaging, and the nosology of Bickerstaff's brainstem encephalitis. PMID- 11422434 TI - Epilepsia partialis continua and frontal features as a debut of anti-Hu paraneoplastic encephalomyelitis with focal frontal encephalitis. PMID- 11422435 TI - Rivastigmine in subcortical vascular dementia: a comparison trial on efficacy and tolerability for 12 months follow-up. PMID- 11422436 TI - Intracerebral haemorrhage under interferon-beta therapy. PMID- 11422437 TI - Isolated muscular sarcoidosis. PMID- 11422438 TI - Chronic lymphocytic leukemia complicated by progressive multifocal leukoencephalopathy without apparent immunodepression. PMID- 11422439 TI - Cognitive function and Epworth Sleepiness Scale in 'minimal' hepatic encephalopathy. PMID- 11422440 TI - How much brain is necessary for vision? PMID- 11422441 TI - Celiac disease associated with recurrent stroke: a coincidence or cerebral vasculitis? PMID- 11422442 TI - Comparison of the aggregation properties, secondary structure and apoptotic effects of wild-type, Flemish and Dutch N-terminally truncated amyloid beta peptides. AB - The Dutch (E22Q) and Flemish (A21G) mutations in the betaAPP region of the amyloid precursor protein (APP) are associated with familial forms of Alzheimer dementia. However, patients with these mutations express substantially different clinical phenotypes. Therefore, secondary structure and cytotoxic effects of the three Abeta(12-42) variants [wild-type (WT), Dutch and Flemish] were tested. At a concentration of 5 microM the aggregation of these peptides followed the order: Abeta(1-42) WT > Abeta(12-42) WT > Abeta(12-42) Flemish > Abeta(12-42) Dutch. The stability of the secondary structure of these peptides upon decreasing the trifluoroethanol (TFE) concentration in the buffer was followed by circular dichroism measurements. WT peptides progressively lost their alpha-helical structure; this change occurred faster for both the Flemish and Dutch peptides, and at higher percentages of TFE in the buffer, and was accompanied by an increase in beta-sheet and random coil content. Apoptosis was induced in neuronal cells by the Abeta(12-42) WT and Flemish peptides at concentrations as low as 1-5 microM, as evidenced by propidium iodide (PI) staining, DNA laddering and caspase 3 activity measurements. Even when longer incubation times and higher peptide concentrations were applied the N-truncated Dutch peptide did not induce apoptosis. Apoptosis induced by the full length Abeta(1-42) peptide was weaker than that induced by its N-truncated variant. These data suggest that N truncation enhanced the cytotoxic effects of Abeta WT and Flemish peptides, which may play a role in the accelerated progression of dementia. PMID- 11422443 TI - NMDA receptor subunits GluRepsilon1, GluRepsilon3 and GluRzeta1 are enriched at the mossy fibre-granule cell synapse in the adult mouse cerebellum. AB - Cerebellar N-methyl-D-aspartate (NMDA) receptors are concentrated in the granular layer and are involved in motor coordination and the induction of long-term potentiation at mossy fibre-granule cell synapses. In the present study, we used immunohistochemistry to examine the distribution of NMDA receptor subunits in the adult mouse cerebellum. We found that appropriate pepsin pretreatment of sections greatly enhanced the sensitivity and specificity of immunohistochemical detection. As a result, intense immunolabelling for GluRepsilon1 (NR2A), GluRepsilon3 (NR2C), and GluRzeta1 (NR1) all appeared in synaptic glomeruli of the granular layer. Double immunofluorescence showed that these subunits were colocalized in individual synaptic glomeruli. Within the glomerulus, NMDA receptor subunits were located between centrally-located huge mossy fibre terminals and peripherally-located tiny Golgi axon terminals. By immunoelectron microscopy, all three subunits were detected at the postsynaptic junction in granule cell dendrites, forming synapses with mossy fibre terminals. Consistent with the known functional localization, GluRepsilon1, GluRepsilon3, and GluRzeta1 are, thus, anatomically concentrated at the mossy fibre-granule cell synapse. By contrast, immunohistochemical signals were very low in Purkinje cell somata and dendrites in the molecular layer. The lack of GluRzeta1 immunolabelling in Purkinje cells was unexpected because the cells express GluRzeta1 mRNA at high levels and high levels of GluRzeta1 protein in the molecular layer were revealed by immunoblot. As Purkinje cells are exceptionally lacking GluRepsilon expression, the discrepant result may provide in vivo evidence suggesting the importance of accompanying GluRepsilon subunits in synaptic localization of GluRzeta1. PMID- 11422444 TI - Ceramide-induced apoptosis in cortical neurons is mediated by an increase in p38 phosphorylation and not by the decrease in ERK phosphorylation. AB - Ceramide, the central molecule of the sphingomyelin pathway, serves as a second messenger for cellular functions ranging from proliferation and differentiation to growth arrest and apoptosis. In this study we show that c2-ceramide induces apoptosis in primary cortical neuron cultures and that this effect correlates with differential modulation of mitogen-activated protein kinase (MAPK) cascades. Phosphorylation of extracellular signal-regulated kinases (ERKs) and their upstream activators MAPK kinases (MEKs), as measured by immunoblotting is rapidly decreased by c2-ceramide. However, the MEK inhibitor PD98059 alone does not induce apoptosis and in combination with c2-ceramide it does not modify c2 ceramide-induced apoptosis. Treatment with c2-ceramide increases p38 and c-Jun N terminal kinase (JNK) phosphorylation before and during caspase-3 activation. The p38 inhibitor SB203580 partially protects cortical neurons against c2-ceramide induced apoptosis, implicating the p38 pathway in this process. The c2-ceramide treatment also increases levels of c-jun, c-fos and p53 mRNA in primary cortical neuron cultures, but this is independent of p38 activation. Our study further elucidates the time-courses of MAPK cascade modulation, and of c-jun, c-fos and p53 activation during c2-ceramide-induced neuronal apoptosis. It reveals that one of the activated kinases, p38, is necessary for this apoptosis. PMID- 11422445 TI - ATP but not adenosine inhibits nonquantal acetylcholine release at the mouse neuromuscular junction. AB - The postsynaptic membrane of the neuromuscular synapse treated with antiacetylcholinesterase is depolarized due to nonquantal release of acetylcholine (ACh) from the motor nerve ending. This can be demonstrated by the hyperpolarization produced by the application of curare (H-effect). ATP (1 x 10-5 M) decreased the magnitude of the H-effect from 5 to 1.5 mV. The membrane input resistance and the ACh sensitivity were unchanged, and so changes in these cannot explain the ATP effect. Adenosine alone was without effect on the nonquantal release. On the other hand, both ATP and adenosine depressed the frequency of spontaneous miniature endplate potentials, to 56% and 43% respectively. The protein kinase A inhibitor Rp-cAMP or the guanylyl cyclase inhibitor 1H [1,2,4]oxidiazolo[4,3-a]quinoxalin-1-one did not affect the inhibitory influence of ATP on the H-effect, whereas staurosporine, an inhibitor of protein kinase C, completely abolished the action of ATP. Suramin, an ATP antagonist, enhanced the H-effect to 8.6 mV and, like staurosporine, prevented the inhibitory effect of ATP. ATP thus suppresses the nonquantal release via a direct action on presynaptic metabotropic P2 receptors coupled to protein kinase C, whilst adenosine exerts its action mainly by affecting the mechanisms underlying quantal release. These data, together with earlier evidence, show that nonquantal release of ACh can be modulated by several distinct regulatory pathways, in particular by endogenous substances which may or may not be present in the synaptic cleft at rest or during activity. PMID- 11422446 TI - DNA elements of the type 1 adenylyl cyclase gene locus enhance reporter gene expression in neurons and pinealocytes. AB - The Ca2+-stimulated type 1 adenylyl cyclase (AC1) contributes to several forms of synaptic plasticity and is the only known neurospecific adenylyl cyclase. Furthermore, the protein and mRNA levels of AC1 undergo a circadian oscillation in the pineal gland, and AC1 may play a pivotal role in regulating nocturnal melatonin synthesis. To better understand the expression of AC1, we isolated mouse genomic DNA clones of AC1. The transcription and translation start regions of mouse AC1 share extensive homologies with the bovine counterpart. The upstream proximal region has potential binding sites for transcription factors, including the steroid receptor family, the E-box factors, and Sp1. A 280-bp fragment that contains the transcription start site directed reporter gene expression in cultured cortical neurons and pinealocytes functioning as a basal neuro- and pineal-directed promoter. Interestingly, pinealocyte expression of the reporter gene was inhibited by increases in cAMP. This cAMP sensitivity may explain why AC1 mRNA in the pineal is low at night when cAMP is elevated and high during the day when cAMP signals drop. An adjacent 330-bp fragment interacted specifically with nuclear factor(s) that we designate binary E-box factor (BEF). Methylation interference and DNase I footprinting identified the BEF-binding site sequence as 5'-CCAAGGTCACGTGGC-3'. When linked to the basal tissue-directed promoter, this 15 bp sequence further enhanced reporter expression in neurons and pinealocytes. We propose that this 15-bp sequence may contribute to increased expression of AC1 in neurons and pinealocytes relative to other cells. PMID- 11422447 TI - Antiproliferative properties of sphingosine-1-phosphate in human hepatic myofibroblasts. AB - Sphingosine-1-phosphate (S1P) is a potent lysophospholipid mediator mostly released by activated platelets. It is involved in several functions in peripheral tissues, but its effects in the central nervous system are poorly documented. Therefore, we have examined the effects of S1P on the proliferation of striatal astrocytes from the mouse embryo. These cells have been found to express mRNAs for the S1P receptors, Edg-1 and Edg-3. S1P stimulated thymidine incorporation and induced activation of extracellular signal-regulated kinases (Erks). Both effects were prevented by U0126, an Erk kinase inhibitor. The S1P evoked activation of Erk1 was totally blocked in astrocytes pretreated with a combination of either phorbol ester (24 h) and LY294002, or phorbol ester (24 h) and pertussis toxin (PTX). Each individual treatment only partially inhibited Erk1 activation. This suggests that several separate mechanisms mediate this process, one involving protein kinase C and another involving Gi/Go proteins and phosphatidylinositol 3-kinase. In contrast, the stimulatory effect of S1P on astrocyte proliferation was totally blocked by either PTX or LY294002, but not by a downregulation of protein kinase C. S1P dramatically inhibited the evoked production of cyclic AMP, a response that was impaired by PTX. Finally, S1P stimulated the production of inositol phosphates and increased intracellular calcium by mobilization from thapsigargin-sensitive stores. These latter effects were mainly insensitive to PTX. Probably, Gi/Go protein activation and phosphoinositide hydrolysis are early events that regulate the activation of Erks by S1P. Altogether, these observations show that astrocytes are targets for S1P. Their proliferation in response to S1P could have physiopathological consequences at sites of brain lesions and alterations of the blood-brain barrier. PMID- 11422448 TI - Effect of the selective noradrenergic reuptake inhibitor reboxetine on the firing activity of noradrenaline and serotonin neurons. AB - Reboxetine is a non-tricyclic antidepressant with selective noradrenergic (NA) reuptake-blocking effects. The effects of acute and sustained administration of reboxetine, on the firing activity of locus coeruleus NA neurons and dorsal raphe 5-HT neurons, were assessed using in vivo extracellular unitary recording in rats anaesthetized with chloral hydrate. Reboxetine (0.1-1.25 mg/kg, i.v.) dose dependently decreased the firing activity of NA neurons (ED50 = 480 +/- 14 microg/kg). A 2-day treatment with reboxetine at 1.25, 2.5, 5, or 10 mg/kg per day (using osmotic minipumps implanted subcutaneously) produced significant decreases of 52%, 68%, 81%, and 83%, respectively, of NA firing activity. When the reboxetine treatment (2.5 mg/kg per day) duration was prolonged to 7 days, a 66% decrease in NA firing activity was observed which further decreased to 80% after 21 days of treatment. In contrast, 5-HT neuron firing rate remained unaltered following short- and long-term reboxetine treatments. The suppressant effect of the alpha2-adrenoceptor agonist clonidine on the firing activity of NA neurons was unchanged in long-term reboxetine-treated rats, but its effect on the firing activity of 5-HT neurons was blunted. The enhancement of NA firing activity by the 5-HT1A agonist 8-OH-DPAT was abolished in long-term reboxetine treated rats, whereas, the inhibitory effect of the 5-HT2 agonist DOI was attenuated by about three-fold. In conclusion, sustained NA reuptake blockade by reboxetine lead to profound alterations in the function of NA neurons and of 5-HT receptors modulating their firing activity. PMID- 11422449 TI - Slow IPSC kinetics, low levels of alpha1 subunit expression and paired-pulse depression are distinct properties of neonatal inhibitory GABAergic synaptic connections in the mouse superior colliculus. AB - Remodelling of visual maps in the superior colliculus (SC) depends on neuronal activity. Synaptic inhibition could contribute to this process because spontaneous spike discharge in the SC was modulated by GABA(A) receptor activation at postnatal days (P) 1-3. To investigate the functional capacity of GABAergic synaptic transmission at this early stage of development, whole-cell patch-clamp recordings were made from wide field neurons (WFNs) in horizontal slices comprising the superficial grey layer of the SC. Focal stimulation in the vicinity of WFNs evoked tetrodotoxin-sensitive stimulus-locked inhibitory postsynaptic currents (eIPSCs). The failure rate of eIPSCs was low ( approximately 0.2), and the maximal amplitude of evoked unitary eIPSCs exceeded the amplitude of average miniature IPSCs (mIPSCs) by a factor of 4-5, suggesting that action potential-mediated GABA release was more effective than spontaneous release. Some of the properties of GABAergic synaptic transmission in the neonatal SC were age-specific. In contrast with eIPSCs in the more mature SC at P20-22, neonatal eIPSCs decayed more slowly, preferentially fluctuated in duration, not amplitude, and mostly lacked temporal summation, due to depression at shorter intervals. The paired-pulse ratio (eIPSC2 : eIPSC1) was inversely related to the duration of eIPSCs. PCR analysis showed, in addition, that the ratio of alpha1 : alpha3 subunit expression was lower in the neonatal SC. Together, these results suggest that, at a young age, efficacy of GABAergic synaptic transmission is primarily constrained by the slow kinetics and the saturation of postsynaptic GABA(A) receptors. PMID- 11422450 TI - Colocalization of CGRP with 5-HT1B/1D receptors and substance P in trigeminal ganglion neurons in rats. AB - Vasodilatation in the dura mater has been implicated in migraine pathogenesis. Anti-migraine triptan drugs block vasodilatation by binding to 5-HT1B/1D receptors localized on the peripheral sensory terminals and dural blood vessel smooth muscles. Previous studies suggest that calcitonin gene-related peptide (CGRP) released from Adelta-fibres plays a more important role than substance P (SP) released from C-fibres in inducing dural vasodilatation and that one of the antimigraine mechanisms of triptan drugs is inhibiting CGRP release. In the present study, the relationship between CGRP and 5-HT1B/1D receptors, and between CGRP and SP in the trigeminal ganglion neurons in rats was examined by double immunohistochemical staining. CGRP, 5-HT1B, 5-HT1D and SP-positive trigeminal ganglion neurons were all predominantly small and medium-sized. In the trigeminal ganglia, approximately 50% of CGRP-positive neurons were 5-HT1B positive. Similarly, approximately 55% of CGRP-positive neurons were 5-HT1D immunoreactive. Approximately 50% of CGRP-positive neurons were SP-positive, while 93% of SP positive neurons were CGRP-positive, suggesting that nearly all SP-positive neurons also contain CGRP. The fibre types of the 5-HT1B- and 5-HT1D-positive neurons were further investigated with an antibody against the A-fibre marker 200 kDa neurofilaments (NF200). Approximately 46% of the 5-HT1B-positive and 43% of the 5-HT1D-positive trigeminal ganglion neurons were also NF200 positive, indicating that many A-fibre trigeminal neurons express 5-HT1B or 5-HT1D receptors. These results support the hypothesis that one important action of antimigraine drugs is the inhibition of CGRP release and that Adelta-fibres may play an important role in migraine pathogenesis. PMID- 11422451 TI - VR1 protein expression increases in undamaged DRG neurons after partial nerve injury. AB - Changes in phenotype or connectivity of primary afferent neurons following peripheral nerve injury may contribute to the hyperalgesia and allodynia associated with neuropathic pain conditions. Although earlier studies using partial nerve injury models have focused on the role of damaged fibres in the generation of ectopic discharges and pain, it is now thought that remaining undamaged fibres may be equally important. We have examined the expression of the sensory neuron-specific cation channel Vanilloid Receptor 1 (VR1), an important transducer of noxious stimuli, in three models of nerve injury in the rat, using anatomical separation or fluorescent retrograde tracers to identify damaged or undamaged sensory neurons. After total or partial sciatic nerve transection, or spinal nerve ligation, VR1-immunoreactivity (IR) was significantly reduced in the somata of all damaged dorsal root ganglion (DRG) neuronal profiles, compared to controls. However, after partial transection or spinal nerve ligation, VR1 expression was greater in the undamaged DRG somata than in controls. Unexpectedly, after L5 spinal nerve ligation, VR1-IR of the A-fibre somata increased approximately 3-fold in the uninjured L4 DRG compared to controls; a much greater increase than seen in the somata with C-fibres. Furthermore, we found that VR1-IR persisted in the transected sciatic nerve proximal to the lesion, despite its down-regulation in the damaged neuronal somata. This persistence in the nerve proximal to the lesion after nerve section, together with increased VR1 in DRG neurons left undamaged after partial nerve injury, may be crucial to the development or maintenance of neuropathic pain. PMID- 11422452 TI - Potentiation of 3-hydroxyglutarate neurotoxicity following induction of astrocytic iNOS in neonatal rat hippocampal cultures. AB - Neuronal damage in glutaryl-CoA dehydrogenase deficiency (GDD) has previously been addressed to N-methyl-D-aspartate (NMDA) receptor-mediated neurotoxicity of the accumulating neurotoxic metabolite 3-hydroxyglutarate. However, acute encephalopathic crises in GDD patients are typically precipitated by febrile illness or even routine vaccinations, suggesting a potentiating role of inflammatory cytokines. In the present study we investigated the effect of interleukin-1beta and interferon-gamma on 3-hydroxyglutarate toxicity in rat cortical astrocyte cultures and neonatal rat hippocampal cultures. A cotreatment of both culture systems with interleukin-1beta and interferon-gamma induced the protein expression of astrocytic inducible nitric oxide synthase (iNOS), resulting in increased nitric oxide (NO) production. Cytokine pretreatment alone had no effect on cell viability but potentiated 3-hydroxyglutarate neurotoxicity. NOS inhibition by aminoguanidine and L-NAME prevented an iNOS-mediated potentiation of 3-hydroxyglutarate neurotoxicity but failed to protect neurons against 3-hydroxyglutarate alone. In contrast, superoxide dismutase/catalase as well as MK-801 prevented toxicity of 3-hydroxyglutarate alone as well as its potentiation by iNOS, supporting a central role of NMDA receptor stimulation with subsequently increased superoxide anion production. It is concluded that the potentiation of 3-hydroxyglutarate neurotoxicity is most probably due to an induction of astrocytic iNOS and concomitantly increased NO production, enabling enhanced peroxynitrite formation. Thus, we provide evidence for a neuroimmunological approach to the precipitation of acute encephalopathic crises in GDD by inflammatory cytokines. PMID- 11422453 TI - Distribution of mossy fibre rosettes in the cerebellum of cat and mice: evidence for a parasagittal organization at the single fibre level. AB - Mossy fibres are the main afferent input to the granular layer of the cerebellar cortex. In this study, the spatial distribution of the mossy fibres' presynaptic enlargements - the so-called rosettes - were analysed on the single fibre level. Data obtained from the cerebella of cat and mice were compared to look for species differences, and the cerebella of the adult and young mice were also compared to look for developmental changes. The results show that there is a spatial anisotropy in all mossy fibres studied, with neighbouring rosettes being about three times further away from each other along the parasagittal axis and closer to each other in the mediolateral direction. Furthermore, these results suggest that this anisotropy is established at an early developmental stage. The anisotropic orientation of mossy fibres at the single fibre level supports the hypothesis of a timing mechanism in cerebellar function. PMID- 11422454 TI - Oligodendroglial tau filament formation in transgenic mice expressing G272V tau. AB - Genetic evidence indicates that several mutations in tau, including G272V, are linked to frontotemporal dementia with parkinsonism. We expressed this mutation in mouse brains by combining a prion protein promoter-driven expression system with an autoregulatory transactivator loop that resulted in high expression of human G272V tau in neurons and in oligodendrocytes. We show that G272V tau can form filaments in murine oligodendrocytes. Electron microscopy established that the filaments were either straight or had a twisted structure; these were 17-20 nm wide and had a periodicity of approximately 75 nm. Filament formation was associated with tau phosphorylation at distinct sites, including the AT8 epitope 202/205 in vivo. Immunogold electron microscopy of sarcosyl-extracted spinal cords from G272V transgenic mice using phosphorylation-dependent antibodies AT8 or AT100 identified several sparsely gold-labelled 6-nm filaments. In the spinal cord, fibrillary inclusions were also identified by thioflavin-S fluorescent microscopy in oligodendrocytes and motor neurons. These results establish that expression of the G272V mutation in mice causes oligodendroglial fibrillary lesions that are similar to those seen in human tauopathies. PMID- 11422455 TI - Presynaptic kynurenate-sensitive receptors inhibit glutamate release. AB - Kynurenic acid is a tryptophan metabolite provided with antagonist activity on ionotropic glutamate and alpha7 nicotinic acetylcholine receptors. We noticed that in rats with a dialysis probe placed in the head of their caudate nuclei, local administration of kynurenic acid (30-100 nM) significantly reduced glutamate output. Qualitatively and quantitatively similar effects were observed after systemic administration of kynurenine hydroxylase inhibitors, a procedure able to increase brain kynurenate concentrations. Interestingly, in microdialysis studies, methyllycaconitine (0.3-10 nM), a selective alpha7 nicotinic receptor antagonist, also reduced glutamate output. In isolated superfused striatal synaptosomes, kynurenic acid (100 nM), but not methyllycaconitine, inhibited the depolarization (KCl 12.5 mM)-induced release of transmitter or previously taken up [3H]-D-aspartate. This inhibition was not modified by glycine, N-methyl-D aspartate or subtype-selective kainate receptor agents, while CNQX or DNQX (10 microM), two AMPA and kainate receptor antagonists, reduced kynurenic acid effects. Low concentrations of kynurenic acid, however, did not modify [3H] kainate (high and low affinity) or [3H]-AMPA binding to rat brain membranes. Finally, because metabotropic glutamate (mGlu) receptors modulate transmitter release in striatal preparations, we evaluated, with negative results, kynurenic acid (1-100 nM) effects in cells transfected with mGlu1, mGlu2, mGlu4 or mGlu5 receptors. In conclusion, our data show that kynurenate-induced inhibition of glutamate release is not mediated by glutamate receptors. Nicotinic acetylcholine receptors, however, may contribute to the inhibitory effects of kynurenate found in microdialysis studies, but not in those found in isolated synaptosomes. PMID- 11422456 TI - Conscious recollection and illusory recognition: an event-related fMRI study. AB - In this event-related functional magnetic resonance imaging (fMRI) study we examined the neuronal correlates of the subprocesses underlying recognition memory. In an explicit memory task, participants had to discriminate studied ('old') words from semantically related and unrelated 'new' (unstudied) words. We examined whether the correct rejection of semantically related words which were similar to old words, which had elicited correct responses, was based on conscious recollection of study phase information. In this task, false-positive responses to semantically related new words can be assumed to result from the assessment of the semantic similarity between test words and studied words with minimal recollection. For correct identification of old words and correct rejection of new related words we found common activation in a variety of brain areas that have been shown to be involved in conscious recollection, among them the left middle frontal gyrus, the precuneus, the retrosplenial cortex, the left parahippocampal gyrus and the thalamus. For correct responses to old words, the frontomedian wall, the posterior cingulate cortex and the nucleus accumbens were additionally activated, suggesting an emotional contribution to these judgements. Correct rejections of related new words were associated with additional activation of the right middle frontal gyrus, suggesting higher monitoring demands for these more difficult recognition judgements. False-positive responses to semantically related new words were associated with enhanced activation in the frontomedian wall. The results point to an important role of the prefrontal cortex as well as medial temporal and medial parietal regions of the brain in successful memory retrieval and conscious recollection. PMID- 11422457 TI - The group I metabotropic glutamate receptor antagonist S-4-CPG modulates the locomotor response produced by the activation of D1-like, but not D2-like, dopamine receptors in the rat nucleus accumbens. AB - Functional interactions between dopamine (DA) and glutamate neurotransmissions in both the dorsal and the ventral striatum have been described for long time. However, there is much controversy as to whether glutamate transmission stimulates or attenuates DA release and locomotor activity. We investigated the functional interactions on locomotor activity between group I metabotropic glutamatergic receptors (mGlu receptors) and both D1-like and D2-like DA receptors in the rat nucleus accumbens. Intra-accumbens administration of the selective group I mGlu receptor antagonist S-4-CPG (0.2 or 2 microg per side), which had no effect when injected alone, prevented the increase in locomotor activity produced by the selective D1-like receptor agonist SKF 38393 (1 microg per side). Co-administration with S-4-CPG of the group I mGlu receptor agonist DHPG, but not of the group II mGlu receptor agonist APDC or the group III mGlu receptor agonist AP4, reversed the antagonistic effect of S-4-CPG on the SKF 38393-induced increase in locomotor activity. This indicates that the antagonistic effect of S-4-CPG could result from an action at the group I mGlu receptors. In contrast, administration of S-4-CPG showed no effect on the locomotor responses produced by either the selective D2-like receptor agonist LY 171555 (1 microg per side) or a mixed solution of SKF 38393 + LY 171555 (1 microg per side each). Altogether, these results confirm that glutamate transmission may control locomotor function through mGlu receptors in a DA-dependent manner, and further indicate that group I mGlu receptors would interact with D1-like receptors, but not D2-like receptors, to modulate DA transmission and locomotor activity. PMID- 11422458 TI - Localized immediate early gene expression related to the strength of song learning in socially reared zebra finches. AB - Recent evidence showed that exposure of tape-tutored zebra finch (Taeniopygia guttata castanotis) males to the tutor song involves neuronal activation in brain regions outside the conventional 'song control pathways', particularly the caudal part of the neostriatum (NCM) and of the hyperstriatum ventrale (CMHV). Zebra finch males were reared with a live tutor during the sensitive period for song learning. When, as adults, they were re-exposed to the tutor song, the males showed increased expression of Fos, the protein product of the immediate early gene c-fos, in the NCM and CMHV, compared with expression in two conventional 'song control nuclei', high vocal centre (HVC) and Area X. The strength of the Fos response (which is a reflection of neuronal activation) in the NCM (but not in the other three regions) correlated significantly and positively with the number of song elements that the birds had copied from the tutor song. Thus, socially tutored zebra finch males show localized neural activation in response to tutor song exposure, which correlates with the strength of song learning. PMID- 11422459 TI - Enhanced inactivation and acceleration of activation of the sodium channel associated with epilepsy in man. AB - Generalized epilepsy with febrile seizures-plus (GEFS+) is a benign Mendelian syndrome characterized by childhood-onset febrile and afebrile seizures. Three point mutations within two voltage-gated sodium channel genes have been identified so far: in GEFS+ type 1 a mutation in the beta1-subunit gene SCN1B, and in GEFS+ type 2 two mutations within the neuronal alpha-subunit gene SCN1A. Functional expression of the SCN1B and one of the SCN1A mutations revealed defects in fast channel inactivation which are in line with previous findings on myotonia causing mutations in SCN4A, the skeletal muscle sodium channel alpha subunit gene, all showing an impaired fast inactivation. We now studied the second GEFS+ mutation (T875M in SCN1A), using the highly homologous SCN4A gene (mutation T685M). Unexpectedly, the experiments revealed a pronounced enhancement of both fast and slow inactivation and a defect of channel activation for T685M compared to wild-type channels. Steady-state fast and slow inactivation curves were shifted in the hyperpolarizing direction, entry into slow inactivation was threefold accelerated, recovery from slow inactivation was slowed by threefold and the time course of activation was slightly but significantly accelerated. In contrast to other disease-causing mutations in SCN1A, SCN1B and SCN4A, the only mechanism that could explain hyperexcitability of the cell membrane would be the acceleration of activation. Because the enhancement of slow inactivation was the most obvious alteration in gating found for T685M, this might be the disease causing mechanism for that mutation. In this case, the occurrence of epileptic seizures could be explained by a decrease of excitability of inhibitory neurons. PMID- 11422460 TI - GFRalpha3 is expressed predominantly in nociceptive sensory neurons. AB - Activation of the RET receptor tyrosine kinase by glial-derived neurotrophic factor family members is dependent on a family of coreceptors, GFRalpha1-4. GFRalpha3 preferentially binds the newest member of the glial-derived neurotrophic factor family of ligands, artemin. The major site of GFRalpha3 expression is in the dorsal root ganglion; however, the class of sensory neurons that expresses GFRalpha3 has not been reported previously. Using immunohistochemical methods, we show that the majority of dorsal root ganglion cells that express GFRalpha3 also express vanilloid receptor type 1, peripherin, RET, trkA and calcitonin gene-related peptide. In addition, a significant subpopulation of GFRalpha3-expressing cells also binds the lectin IB4. We demonstrate that GFRalpha3 artemin neurons are immunopositive for markers expected of nociceptors and include a subset of neurons distinct from the GDNF responsive population. Our results indicate artemin may exert selective effects on pain sensation. PMID- 11422461 TI - In search of the Holy Grail of Heliocobacter pylori remedies. AB - In this issue, Gisbert et al. (pp. 157-62) present the results of a noncomparative study evaluating a twice daily, 5-day regimen of ranitidine bismuth citrate, amoxicillin, clarithromycin and metronidazole twice daily for Helicobacter pylori cure. This study is one of a few stuides that evaluate a 5 day triple antimicrobial regimen in combination with a antisecretory agent. Although the study design precludes making any definite conclusion, it does encourage additional investigation of these types of regimens. Randomized controlled trials (RCTs) using regimens containing multiple agents should consider both standard-of-care comparator regimens and comparator regimens that will provide a better understanding of why regimens are more effective or better tolerated. The goal of treatment should be to maintain a lower bound 95% confidence interval (CI) of the point estimate of greater than 80% and a 'delta' (lower bound 95% CI of the difference in rates) of less than 10%. All RCTs should conduct susceptibility testing to evaluate the impact of resistance on efficacy and explain eradication failures. Finally, consideration should be given to the inclusion of patients with functional dyspepsia in H. pylori studies evaluating H. pylori cure since patients with peptic ulcer disease are becoming harder to find. PMID- 11422462 TI - Helicobacter pylori and early duodenal ulcer status post-treatment: a review. AB - BACKGROUND: Data submitted to the FDA were reviewed to analyze the relationship between Helicobacter pylori infection and the incidence of early duodenal ulcers, within 6 weeks, following treatment. MATERIALS AND METHODS: Retrospective analyzes were performed on data from three H. pylori development programs submitted to the FDA: ranitidine-bismuth-citrate (RBC), lansoprazole (L) and omeprazole (O). Efficacy assessments for the RBC, L and O programs were made at end of a 4-week treatment period, 4-6 weeks following the end of a 14-day treatment period, and 4 weeks following the end of a 4-week treatment period, respectively. RESULTS: Overall, there was a 15%, 21% and 23% decrease in the number of patients in the RBC, L and O programs, respectively, with ulcers among H. pylori cleared/eradicated patients post-treatment compared with patients with persistent infection. Among patients who did not have cleared/eradicated H. pylori in the RBC and O programs, where antisecretory agents were continued beyond the antimicrobial treatment period, the number of ulcers was lower in the antisecretory plus antimicrobial subgroups compared with the antimicrobial alone subgroups (37% vs. 46% for RBC and 33% vs. 42% for O). Among patients with cleared/eradicated H. pylori, the number of patients with ulcers in the antimicrobial alone subgroups and antisecretory plus antimicrobial subgroups were similar within each program. Antimicrobials alone had significantly lower rates of ulcers among patients with cleared/eradicated H. pylori as compared with patients without clearance/eradication. CONCLUSIONS: The early incidence of duodenal ulcers is significantly decreased in patients with H. pylori clearance/eradication. PMID- 11422463 TI - Cell lysis is responsible for the appearance of extracellular urease in Helicobacter pylori. AB - BACKGROUND: Helicobacter pylori is a neutralophilic bacterium that colonizes the acidic human gastric surface using the neutralizing capacity of a constitutively produced urease. Urease is present both in the cytoplasm and bound to the outside surface of the bacteria. The origin of the surface urease continues to be controversial. This study provides additional evidence that the origin of surface urease is cell lysis, not secretion. METHODS: H. Pylori was transformed with a plasmid encoding green fluorescent protein (GFP), a non-native cytoplasmic protein. Cultures supplemented with beta-cyclodextrin or horse serum were collected over various time periods and spun through a ficoll cushion to gently separate whole bacteria from released protein. The pellet and supernatant fractions were analyzed by fluorimetry, SDS-PAGE and Coomassie blue or Western analysis. RESULTS: GFP fluorescence and antigenic reactivity in the supernatant increased at each time point. GFP, the non-native cytoplasmic protein, and UreB, a native cytoplasmic protein, increased over time in the supernatant and both proteins were always present in the pellet fraction. UreI, an inner membrane protein, was only present in the pellet fraction. beta-galactosidase, a protein not found in H. pylori, was used as a negative control. CONCLUSIONS: Since it is unlikely that there is an intrinsic secretion system for GFP, a non-native protein, its increasing presence over time in the supernate fraction along with UreB, and retention of UreI in the pellet fraction implies that cell lysis accounts for the presence of urease on the surface of H. pylori. PMID- 11422464 TI - Probing 23S ribosomal RNA cleavage sites in coccoid Helicobacter pylori. AB - BACKGROUND: Previous studies have revealed that extensive nonrandom fragmentation of ribosomal RNA occurs during conversion of Helicobacter pylori to the coccoid form. The 16S rRNA fragmentation has been characterised in some detail. The aim of the present study was to define corresponding cleavage-sites in the 3'-half of the 23S rRNA molecule. MATERIALS AND METHODS: Northern blot analysis using 23S rRNA specific antisense riboprobes and a 5'-end-labelled oligonucleotide probe was used to analyse the 23S rRNA fragmentation pattern in coccoid H. pylori type strain CCUG 17874T and H. pylori 26695, for which the genome has been sequenced. A double-stranded cDNA-dependent (ds-cDNA) primer-extension analysis technique using 23S rRNA ds-cDNA and a primer targeting the vicinity of the peptidyl transferase centre was used to determine cleavage sites at the nucleotide level. RESULTS: We report here the mapping of putative cleavage sites within domains IV and V, enclosing the peptidyl transferase centre, in the 3'-half of the 23S rRNA molecule. Three cleavage sites were located in domain IV. Two other cleavage sites were located in the peptidyl transferase centre, and one presumptive multiple-break site between helices 77 and 78 in domain V. The DNA motifs were different from the postulated A + U rich single-strand cleavage sites recognised by RNase E, which has been implicated in rRNA degradation in Escherichia coli. CONCLUSIONS: The present analysis suggests that a hitherto unknown mechanism is responsible for the nonrandom fragmentation of rRNA in coccoid H. pylori, which may have important consequences for the growth, and survival of the bacterium. PMID- 11422465 TI - Helicobacter pylori induces apoptosis in human epithelial gastric cells by stress activated protein kinase pathway. AB - BACKGROUND: The pathway by which Helicobacter pylori induces apoptosis in gastric epithelial cells is not known. The aim of this study was to determine whether H. pylori-induced apoptosis is associated with SAPK/JNK activity in human gastric cancer KATO III cells. MATERIALS AND METHODS: H. pylori VacA toxin positive strain was incubated with KATO III cells for 0.5, 1, 2 or 24 hours. The SAPK/JNK protein was harvested from the KATO III cell lysate by precipitation with a C-jun fusion protein and its activity was measured by C-jun phosphorylation utilizing transblotting and phosphoserine antibody. Cellular apoptosis was demonstrated by DNA fragmentation. In addition, cell growth in coculture with H. pylori was determined over 72 hours. RESULTS: H. pylori significantly stimulated SAPK/JNK activity in KATO III cells with a peak at the 0.5 hour time point (3.6-fold vs. control, p <.05), but a return to basal levels by 2 hours. In addition, significant DNA fragmentation was observed after 24 hours in these cells but not in the control KATO III cells. Cell growth was inhibited in a dose dependent fashion in coculture with H. pylori. CONCLUSION: These results show that H. pylori triggers an increase in apoptosis in KATO III cells as reflected by DNA fragmentation. This effect was preceded and correlated with an increase in SAPK/JNK activity suggesting that the H. pylori-induced apoptosis in human gastric epithelial cells may be mediated by the SAPK/JNK pathway. PMID- 11422466 TI - Relation between cytokines and Helicobacter pylori in gastric cancer. AB - BACKGROUND: Helicobacter pylori is etiologically involved in the development of gastric cancer and infected gastric mucosa has been shown to possess elevated levels of cytokines [for example interleukin (IL)-1beta, IL-6 and IL-8]. Because specific cytokines have also been shown to enhance the development of certain cancers, we examined the relationship between the levels of cytokines, the type and stage of gastric cancers, and the H. pylori infection. MATERIALS AND METHODS: Cytokines were measured from gastric cancer tissues, adjacent normal appearing mucosa, and the serum in 66 patients with early or advanced gastric cancer and from controls using semiquantitative RT-PCR and ELISA. RESULTS: IL-6 and IL-8 levels were more than 10-fold increased in cancer tissues as compared with normal gastric tissues. IL-8 levels in cancer tissues were more than 2-fold higher in advanced gastric cancer as compared with early gastric cancer irrespective of H. pylori status. IL-6 levels were significantly higher in early gastric cancer with active H. pylori infection as compared with early cancer without H. pylori infection (8.7 + 1.4 vs. 1.2 + 0.3 pg/mg protein, p <.001) and decreased significantly after the cure of H. pylori (11.1 + 2.9-8.2 + 2.3 pg/mg protein, p <.05). CONCLUSIONS: IL-8 levels in gastric cancer tissue are largely independent of H. pylori infection. In contrast, tissue IL-6 levels were high in H. pylori infected early gastric cancer and fell significantly after the cure of H. pylori suggesting a relationship between H. pylori infection and early gastric cancer. PMID- 11422467 TI - Inhibitory action of a novel proton pump inhibitor, rabeprazole, and its thioether derivative against the growth and motility of clarithromycin-resistant Helicobacter pylori. AB - BACKGROUND: Clarithromycin-resistant Helicobacter pylori (CRHP) has increasingly been isolated from patients in Japan. The aim of our study was to test whether proton pump inhibitors (PPIs) and their thioether derivatives, which are secreted into the gastric mucosa, could inhibit the growth and motility (a factor in colonization) of CRHP. MATERIALS AND METHODS: CRHP was isolated from patients who had experienced gastritis or peptic ulcers in Tokyo and Niigata. Drugs and related agents tested were omeprazole, lansoprazole, rabeprazole, the thioether derivative of rabeprazole (rabeprazole-TH), clarithromycin, amoxicillin and metronidazole. The MICs of the drugs and agents for H. pylori strains were determined by the agar dilution METHOD: Bacterial swimming in a liquid layer was examined under an inverted, phase-contrast microscope. RESULTS: The PPIs and rabeprazole-TH, but not the anti-H. pylori agents, inhibited the motility of CRHP at both pH 7.4 and 6.0. The concentrations (microg/ml) necessary to inhibit 50% of the motility at pH 7.4 were 0.25-0.5, 8-32, 8-16 and 128-256 for rabeprazole TH, rabeprazole, lansoprazole and omeprazole, respectively. Rabeprazole-TH exhibited the strongest inhibitory effect against the growth of CRPH (MIC, 0.5 microg/ml). CONCLUSION: Rabeprazole-TH, which is secreted into the gastric mucosa, had the strongest inhibitory action against both the growth and motility of CRHP, suggesting that it is a potential novel agent for CRHP eradication. PMID- 11422468 TI - Discrepancy between polymerase chain reaction assay and Western blot analysis in the assessment of CagA status in dyspeptic patients. AB - BACKGROUND: Infection with CagA-positive Helicobacter pylori may be diagnosed by detecting cagA gene by polymerase chain reaction assay (PCR) or serum antibodies against CagA by Western blot analysis. The aim of this study is to evaluate whether results of PCR and Western blot analysis are in agreement in CagA status assessment. PATIENTS AND METHODS: Thirty-six dyspeptic patients with unknown H. pylori status underwent upper gastrointestinal endoscopy to assess the presence of mucosal lesions and to collect six gastric biopsies (three from the antrum and three from the body) for evaluation of H. pylori infection (rapid urease test, histology and PCR for ureA gene) and gastritis. CagA status was assessed by PCR (cagA gene) on two biopsy specimens and by Western blot analysis of serum (CagA antibodies) in each patient. RESULTS: At endoscopy, nine patients showed normal mucosa, 15 a duodenal ulcer and 12 antral erosions. Twenty-eight patients were found to be H. pylori-positive and eight H. pylori-negative. Of the 28 H. pylori positive patients, 17 were CagA-positive and five were CagA-negative by both methods, five were CagA-positive by Western blot analysis but not by PCR and one was CagA-positive by PCR but not by Western blot analysis. Of the eight H. pylori negative patients, none was CagA-positive by PCR, while six were CagA-positive by Western blot analysis. Therefore, the two tests agreed in only 24 patients (67%). In those patients in whom the PCR and Western blot analysis were not in agreement, the histological features appear to suggest that the results of the Western blot analysis should be considered false positives or false negatives. CONCLUSIONS: PCR and Western blot analysis failed to provide comparable data in many cases. Western blot analysis seems to be more likely to give misleading results than PCR. Thus, PCR seems to be the method of choice to assess CagA status. PMID- 11422469 TI - Interactions among gastric somatostatin, interleukin-8 and mucosal inflammation in Helicobacter pylori-positive peptic ulcer patients. AB - BACKGROUND: To investigate whether Helicobacter pylori infection, but not drugs, affects gastric somatostatin, interleukin-8 (IL-8), histological inflammation through eradication therapy, and interactions among these parameters. METHODS: Twenty-eight H. pylori-positive patients (21 males; mean age 47.0 years) with either gastric ulcer (GU: n = 11) or duodenal ulcer (n = 17) diagnosed endoscopically were treated with dual therapy. Eradication was defined as negative microbiologic tests and 13C-urea breath test. Levels of antral and gastric juice somatostatin and mucosal IL-8 were measured by radioimmunoassay and enzyme-linked immunosorbent assay, respectively. Histology was assessed by the Sydney system. RESULTS: H. pylori was eradicated in 15 patients (10 males, 6 GU) out of 28 (54%). The patients' backgrounds did not affect the eradication of H. pylori. Successes in eradication significantly increased antral and juice somatostatin contents, and dramatically decreased IL-8 levels and histological gastritis. In contrast, persistent H. pylori infection did not affect somatostatin and histological gastritis. An inverse correlation was present between changes in somatostatin levels and histological activity. No relationship was observed in changed values between antral somatostatin and IL-8. CONCLUSIONS: These results indicate that eradication of H. pylori, but not the drugs used, induced an increase in somatostatin levels in the antrum and gastric juice, suggesting a close relationship between H. pylori and gastric somatostatin regulation. A close correlation between an increase in gastric somatostatin levels and the normalization of histological activity was present, suggesting that certain peptide-immune interactions in the gastric mucosa exist in H. pylori infection. PMID- 11422470 TI - Correlation between Helicobacter pylori infection, gastric inflammation and serum homocysteine concentration. AB - BACKGROUND: Epidemiological studies have suggested a link between chronic Helicobacter pylori infection and ischemic heart disease but the underlying mechanism remains elusive. We hypothesized that H. pylori-associated chronic gastritis causes impairment of absorption of vitamin cofactors that are essential in the metabolism of homocysteine and results in hyperhomocysteinemia. MATERIALS AND METHODS: Forty-nine dyspeptic patients were studied. H. pylori infection was defined by rapid urease test and histology. Fasting serum homocysteine level, which was measured by a validated commercial fluorescence polarization immunoassay, was correlated with H. pylori infection statuses and gastric histology. H. pylori-infected patients were followed up for 24 weeks post eradication for changes in serum homocysteine concentration. RESULTS: Univariate analyses showed that serum homocysteine level correlated with increasing age (p <.001), male sex (p =.003) and smoking habit (p =.025). There was no significant difference in serum homocysteine levels between H. pylori infected and uninfected subjects (median 10.5 vs. 10.2 micromol/l). After successful eradication of the bacterium, there was no significant reduction in homocysteine level. Moreover, there was no correlation between homocysteine level and gastric histology including H. pylori density, activity and inflammation scores, presence of atrophy or intestinal metaplasia. CONCLUSIONS: The postulated link between H. pylori infection and ischemic heart disease, if it actually exists, is unlikely to be mediated through hyperhomocysteinemia. PMID- 11422471 TI - Is eradication of Helicobacter pylori with colloidal bismuth subcitrate quadruple therapy safe? AB - BACKGROUND: When standard triple therapy fails to eradicate Helicobacter pylori, quadruple 'rescue' therapy is often used which, in Europe, generally comprises colloidal bismuth subcitrate (CBS) based triple therapy and a proton pump inhibitor. Since hypochlorhydria could greatly increase absorption of the toxic bismuth ion from CBS, we investigated the bismuth status of patients receiving anti-H. pylori quadruple therapy. MATERIALS AND METHODS: In a prospective open label study 34 patients with nonulcer dyspepsia or peptic ulcer disease, who had failed to eradicate H. pylori with standard triple therapy, were subsequently treated with CBS, omeprazole, amoxycillin and metronidazole (BOAM). A further 35 patients received triple therapy for the eradication of H. pylori: CBS, amoxycillin and metronidazole (BAM) (n = 18); placebo bismuth, amoxycillin and metronidazole (AM) (n = 9); or omeprazole, amoxycillin and metronidazole (OAM) (n = 8). Whole blood bismuth levels were determined before and within 24 hours of completing treatment. Analysis of bismuth was by inductively coupled plasma mass spectrometry, and concentrations were compared between groups and with the Hillemand 'alarm level' for blood bismuth (50-100 microg/l). RESULTS: BOAM gave higher blood bismuth levels than BAM (difference in means 13.1, CI 6.0-20.2, p <.001); three (8.8%) patients taking BOAM had concentrations within the Hillemand alarm level at 54.2, 64.7 and 91.8 microg/l. OAM and AM did not alter baseline blood bismuth levels. CONCLUSIONS: Caution should be observed in prescribing CBS with gastric acid suppression, and alternative bismuth preparations should be considered. PMID- 11422472 TI - High efficacy of ranitidine bismuth citrate, amoxicillin, clarithromycin and metronidazole twice daily for only five days in Helicobacter pylori Eradication. AB - AIM: The combination of a proton pump inhibitor (PPI) or ranitidine-bismuth citrate (Rbc) and two antibiotics for 7-10 days are, at present, the preferred treatments in Helicobacter pylori eradication. However, therapies for fewer than 7 days have been scarcely evaluated and it is unknown whether the length of treatment can be shortened, without a lost of efficacy, if three instead of two antibiotics are used. The aim of our study was to evaluate the efficacy of Rbc plus three antibiotics for only 5 days in H. pylori eradication. METHODS: We prospectively studied 80 patients (34% duodenal ulcer, 66% functional dyspepsia) infected by H. pylori. At endoscopy, biopsies were obtained for histological study and rapid urease test, and a 13C-urea breath test was carried out. Urea breath test was repeated 4 weeks after completing eradication treatment with Rbc [400 mg twice a day (bid)], amoxicillin (1 g bid), clarithromycin (500 mg bid) and metronidazole (500 mg bid). All drugs were administered together after breakfast and dinner for 5 days only, and no treatment was administered thereafter. Compliance with therapy was determined from the interrogatory and the recovery of empty envelopes of medications. RESULTS: In 79 out of the 80 patients, H. pylori eradication success or failure was assessed after therapy (one patient was lost from follow-up). All but one of these 79 patients took all the medications (one patient stopped treatment on the day 3 due to nausea/vomiting). Per protocol eradication was achieved in 72/78 (92%; 95% CI, 84 96%) and in 72/80 (90%; 81-95%) by intention-to-treat. Therapy was more effective in patients with duodenal ulcer than in those with functional dyspepsia [100% (87 100%) vs. 85% (73-92%) by intention-to-treat; p <.05]. Adverse effects were described in ten patients (12%), and included the perception of a metallic taste (eight patients), nausea/vomiting (two patients, one of them abandoned the treatment due to this), and diarrhea (two patients). CONCLUSION: The combination of Rbc, amoxicillin, clarithromycin and metronidazole for only 5 days represents a promising therapy for H. pylori infection, due to its high efficacy, simple posology, low cost and excellent tolerance. PMID- 11422473 TI - CagA status in dyspeptic patients with and without peptic ulcer disease in Turkey: association with histopathologic findings. AB - BACKGROUND: CagA seropositivity is closely associated with that of vacuolating cytotoxin (VacA). Helicobacter pylori strains positive for both VacA and CagA were reported to be strongly associated with peptic ulcer disease. Different results reporting that cagA gene is not associated with more serious diseases, lowers the importance of CagA protein as a marker. In this study, CagA seropositivity is examined in Turkish peptic ulcer and nonulcer dyspepsia patients; histopathologic scores of CagA (+) and CagA (-) groups were compared. MATERIALS AND METHODS: Sixty consecutive patients (one gastric ulcer, 13 duodenal ulcer and 46 nonulcer dyspepsia) (mean age 40.9 +/- 14.7; 33 women, 27 men) with dyspeptic complaints who underwent upper gastrointestinal (GI) endoscopy were included. Biopsies from the antrum and corpus were used for histopathologic examination and for rapid urease test. H. pylori-negative patients comprised the control group. Histopathologic findings were graded using a previously described grading system (for inflammation, activity, atrophy, intestinal metaplasia and H. pylori, grades from 0 to 3 were used to quantify the findings). In H. pylori positive patients, antibodies against CagA protein were determined using an ELISA METHOD: RESULTS: H. pylori was (+) in 46 patients. One duodenal ulcer and 13 nonulcer dyspepsia patients were negative for H. pylori. CagA positivity is significantly higher in peptic ulcer patients [12/12] than in nonulcer dyspepsia patients [25/33]. While inflammation, activity and atrophy scores were significantly higher in CagA positive patients, intestinal metaplasia and H. pylori load scores were not. Although the histopathologic scores in controls were lower than CagA (-) group, statistical significance was observed only in inflammation and intestinal metaplasia scores. CONCLUSION: Development of more prominent gastritis and severe atrophy in CagA (+) patients is an indicator of the importance of CagA rather than H. pylori load. Therefore, we suggest that nonulcer dyspepsia patients should also be tested for CagA status along with the tests for H. pylori status; and a positive CagA testing should be considered as an indication for eradication treatment. If CagA is negative, further assesment should be performed to decide whether or not to treat the patient. PMID- 11422474 TI - Helicobacter pylori in school children from the Western province of Sri Lanka. AB - BACKGROUND: Little is known about the prevalence of Helicobacter pylori in Sri Lanka and nothing is known about its prevalence in children. Therefore the prevalence of H. pylori in a group of school children in Sri Lanka was determined. MATERIALS AND METHODS: The presence of H. pylori colonisation was determined by detection of faecal antigen and salivary antibody (IgG) by enzyme immuno assay, in 184 children aged between 5 and 19 years, in the Western Province-Colombo district of Sri Lanka. RESULTS: Overall, only 12/184 (6.5%) had detectable H. pylori antigen in their stools and were considered infected with H. pylori, while 51/184 (27.7%) had H. pylori IgG in saliva. H. pylori salivary IgG declined with age while H. pylori antigen detection increased with age. H. pylori infection, as determined by salivary antibody (66%), was greater in children living in overcrowded conditions, although this was not statistically significant. CONCLUSION: The prevalence of H. pylori among school children in Sri Lanka was 6.5% by detection of faecal antigen and 27.7% by detection of salivary antibody, respectively. Initial infection with H. pylori appeared to occur in early childhood whilst active disease began in late childhood. Overcrowding appears to facilitate the transmission of the organism. Overall the prevalence of H. pylori was low in Sri Lanka compared with other countries in South-east Asia. PMID- 11422475 TI - Intra-abdominal spindle cell lesions: a review and practical aids to diagnosis. AB - Intra-abdominal spindle cell lesions are uncommon and often present a diagnostic challenge. An important group of such lesions are the gastrointestinal stromal tumours. Other intra-abdominal spindle cell lesions include fibromatosis, various sarcomas-in particular, leiomyosarcoma, liposarcoma, and malignant peripheral nerve sheath tumour-and, in women, endometrial stromal sarcoma. Less common lesions are inflammatory myofibroblastic tumours, the mesenteric spindle cell reactive lesions, retroperitoneal fibrosis, and solitary fibrous tumour. A variety of intra-abdominal tumours of nonmesenchymal origin may have a spindle cell/sarcomatoid morphology; these include sarcomatoid carcinoma, malignant melanoma and, in women, sarcomatoid granulosa cell tumour. Finally, metastatic sarcomas from pelvic or extra-abdominal organs need also be considered. A set of practical aids to the diagnosis of intra-abdominal spindle cell lesions is presented to assist pathologists dealing with such lesions, particularly with regards to the consideration of differential diagnoses. PMID- 11422476 TI - Immunohistochemical staining for calretinin is useful in the diagnosis of ovarian sex cord-stromal tumours. AB - AIMS: Ovarian sex cord-stromal tumours are a heterogeneous group of neoplasms which may be confused morphologically with a wide variety of tumours. Calretinin positivity has previously been demonstrated in a small number of ovarian sex cord stromal tumours. The aim of this study was to investigate calretinin staining in a series of these tumours and their histological mimics in order to determine the value of calretinin staining in a diagnostic setting. METHODS AND RESULTS: Seventy-two neoplasms, including 37 ovarian sex cord-stromal tumours and 35 miscellaneous neoplasms which may enter into the differential diagnosis, were stained with a commercially available polyclonal antibody against calretinin. All sex cord-stromal tumours exhibited positivity except for a single fibrothecoma. In this group of tumours staining was generally diffuse and strong. Small numbers of the miscellaneous group of neoplasms exhibited positivity but this tended to be focal and weak, although this was not always the case. There was consistent strong positive staining of granulosa cells in follicular cysts and corpora lutea. There was also positive staining of luteinized stromal cells in two cases of ovarian stromal hyperplasia and hyperthecosis. CONCLUSIONS: Calretinin is a sensitive immunohistochemical marker of ovarian sex cord-stromal tumours and may be useful in a diagnostic setting. However, the value is somewhat limited since occasional neoplasms which enter into the morphological differential diagnosis may be positive. Be that as it may, calretinin positivity may be of value in the diagnosis of an ovarian sex cord-stromal tumour and its differentiation from other neoplasms. In this regard, calretinin should always be used as part of a larger panel. PMID- 11422477 TI - Over-diagnosis of hydatidiform mole in early tubal ectopic pregnancy. AB - AIMS: Tubal ectopic hydatidiform moles are rare lesions, and only 40 cases have been reported in the world literature. We investigated the apparently high incidence of tubal ectopic hydatidiform moles in women referred for treatment to a Supraregional Trophoblastic Tumour Screening and Treatment Centre between 1986 and 1996. METHODS AND RESULTS: Of 4261 women referred during the study period, 25 (0.6%) had a suspected tubal ectopic hydatidiform mole and paraffin-embedded tissue was available in 20 (80%) of these. Each case was reviewed by two pathologists and DNA flow cytometric analysis was undertaken when the histological diagnosis was initially deemed equivocal or suggestive of hydatidiform mole. On review, 17 cases (85%) showed no evidence of hydatidiform mole (circumferential trophoblastic proliferation, hydrops, scalloped villi, and stromal karyorrhexis). Of these, 11 cases (65%) showed features of early placentation and six (35%) showed hydropic abortion. DNA flow cytometry was performed in 14 (82%) of these cases and revealed a diploid population in each case. Three cases of molar pregnancy (15%) were identified. Each of these cases had the histological features of an early complete hydatidiform mole. Sufficient tissue was available for DNA flow cytometric analysis in two of these cases and confirmed the presence of diploidy in each. CONCLUSION: Our results show that tubal ectopic hydatidiform mole is a rare entity and demonstrate that it is over diagnosed. Polar trophoblastic proliferation and hydropic villi are features of early placentation and of hydropic abortion. Sheets of extravillous trophoblast may be particularly prominent in tubal ectopic gestation. In the absence of circumferential trophoblastic proliferation combined with hydropic change a diagnosis of gestational trophoblastic disease should be avoided. PMID- 11422478 TI - Specific staining of human chorionic gonadotropin beta in benign and malignant gastrointestinal tissues with monoclonal antibodies. AB - AIMS: Human chorionic gonadotropin (hCG) beta in serum is a promising tumour marker for gastrointestinal malignancies. Our aim was to investigate the expression of hCGbeta by immunohistochemistry in various gastrointestinal cancers and benign tissues. METHODS AND RESULTS: A monoclonal antibody (MAb) specific for free hCGbeta was used to stain 107 tissue samples from various gastrointestinal malignancies and 36 benign or normal tissue samples. The specificity of the staining was verified and the results compared with those obtained with a widely used commercial polyclonal antibody (PAb) which reacts with both free hCGbeta and intact hCG, as well as with luteinizing hormone beta. With the MAb, we observed positive immunohistochemical staining in 24% of the malignant gastrointestinal tumours. Gastric (60%) and pancreatic (56%) carcinomas, as well as extrahepatic cholangiocarcinomas (36%), were positive most frequently. We also discovered immunoreactivity in half of the non-malignant samples from pancreatic and biliary tissues. With the PAb, hCG immunoreactivity was evident more frequently in some cancers, but the staining was diffuse and occasionally polymorphonuclear leucocytes were strongly stained. CONCLUSIONS: This study shows that our MAbs specific for hCGbeta are well suited for immunohistochemistry. Our results confirm previous findings on gastrointestinal cancers and, furthermore, we demonstrate hCGbeta tissue expression in pancreatic adenocarcinoma. The results support reports on hCGbeta as a serum tumour marker for digestive tract diseases. PMID- 11422479 TI - Lung carcinoma with rhabdoid cells: a clinicopathological study and survival analysis of 14 cases. AB - AIMS: We determined the clinicopathological features of primary lung carcinomas with rhabdoid cells by defining the immunophenotype of rhabdoid cells and analysing survival. METHODS AND RESULTS: Rhabdoid cells are distinctive in having an eccentric nucleus and a large intracytoplasmic inclusion on routinely stained sections. Based on the number of rhabdoid cells, 45 cases of large cell carcinoma were divided into the following three types: lung tumour with a rhabdoid phenotype (LTRP) (n=4), lung carcinoma with a small number of rhabdoid cells (LCSR) (n=10), large cell carcinoma containing no rhabdoid cells (LCNR) (n=31). LTRP is composed of at least 10% rhabdoid cells. In LCSR the percentage of rhabdoid cells is less than 10%. LTRP and LCSR are associated with locally advanced disease. Immunohistochemical stains were positive for epithelial markers in all LTRP and eight LCSR, for neuroendocrine markers in one LTRP and three LCSR. The outcome is worse for patients with LTRP than LCSR or LCNR. LCSR shows a trend close to LCNR. Stage-matched survival analysis, however, revealed no statistically significant difference among the histological subtypes. CONCLUSIONS: Rhabdoid cells are heterogeneous except for epithelial markers and vimentin positivity. Less than 5% of rhabdoid cells has a negligible effect on prognosis. PMID- 11422480 TI - Pulmonary artery dissection in patients without underlying pulmonary hypertension. AB - AIMS: Pulmonary artery (PA) dissection is a rare event which usually occurs in patients with underlying pulmonary hypertension. We describe two patients who developed PA dissection without pre-existing pulmonary hypertension and present an extensive review of the literature. METHODS AND RESULTS: In the first patient (a 59-year-old woman), acute-onset dyspnoea was initially thought to have been caused by pulmonary thrombosis, and thromboendarterectomy was performed. Histologically, pulmonary dissection without external rupture was evident, chiefly in the right main PA. In the second patient, an 85-year-old man who had hypergammaglobulinaemia of unknown cause and died from a haemorrhagic gastric ulcer, arterial dissection was detected at autopsy. There was no underlying pulmonary hypertension in either patient. Although the true reason for the development of dissection is unclear, pre-existing inflammation was considered to be related to its formation, at least in the second case. CONCLUSIONS: A literature review indicated that idiopathic and inflammation-related PA dissection is extremely unusual. Since PA dissection is very rare, it is important to be aware of its features in order to make a correct diagnosis. PMID- 11422481 TI - Lipomembranous fat necrosis in three cases of testicular torsion. AB - AIMS: To describe for the first time a lesion termed lipomembranous fat necrosis (LFN) in three patients with spermatic cord torsion. METHODS AND RESULTS: We reviewed 386 testes and their epididymides and spermatic cords which had been removed for testicular infarction. For the three cases showing LFN, a battery of histochemical tests (including periodic acid-Schiff (PAS), orcein, Sudan black and Perls stains) was applied and clinical histories and laboratory data were also investigated. Findings were similar in the three specimens. The testes showed a central group of necrotic seminiferous tubules which were surrounded by granulation tissue consisting of macrophages, multinucleated giant cells, lymphocytes, plasma cells and fibrous connective tissue at the periphery of the lesion. The spermatic cord showed thrombosed veins surrounded by fat necrosis showing cystic cavities which were bounded by wavy hyaline membranes. These stained with Sudan black, PAS (before and after diastase digestion) and orcein and presented yellowish-green autofluorescence. CONCLUSIONS: Lipomembranous fat necrosis of the spermatic cord is a distinctive entity which seems to be related to spermatic cord torsion and the differential diagnosis of which should be established with regard to the presence of parasites, sclerosing lipogranuloma and granuloma evoked by rupture of a testicular prosthesis. PMID- 11422482 TI - Spitzoid malignant melanoma in teenagers: an entity with no better prognosis than that of other forms of melanoma. AB - AIMS: A rare form of melanoma in teenagers closely simulates Spitz naevus and is claimed to have a good prognosis. The aim of this study is to identify the clues for a confident diagnosis of this entity and to confirm the peculiarly good prognosis. METHODS AND RESULTS: Two cases of melanoma with Spitzoid features were compared with Spitz naevus and it was found that the major distinctive criteria are: mitoses and single cell necrosis in the deepest part of the lesion, cellular and particularly nuclear and nucleolar pleomorphism, and growth pattern in solid sheets of cells. More subtle clues were the asymmetric distribution of pigment and the thinning of the epidermis with parakeratosis and exudate in the cornified layer. Both of the lesions reached the mid-dermis. There was a fatal outcome in both patients after generalized metastatic spread. The metastatic disease in one of the cases appeared 15 years after the excision of the primary lesion. CONCLUSIONS: Spitzoid melanoma in teenagers can be distinguished from Spitz naevus if strict criteria are followed. Spitzoid melanoma does not show a better prognosis than other types of melanoma if the follow-up is prolonged enough. PMID- 11422483 TI - Proliferating trichilemmal tumour: p53 immunoreactivity in association with p27Kip1 over-expression indicates a low-grade carcinoma profile. AB - AIMS: Alterations of cell-cycle regulatory molecules in tumorigenesis may predict the biological behaviour of neoplasms and greatly contribute to their proper classification. Since the behaviour of proliferating trichilemmal tumour (PTT) is controversial, we decided to explore the possible significance of altered p53 and p27Kip1 immunohistochemical expression patterns in PTT. METHODS AND RESULTS: We evaluated the percentage and distribution of positive tumour cells and compared the results with those obtained from usual trichilemmal cysts (TC) and squamous cell carcinomas with trichilemmal differentiation (SCCT). PTT showed p53 immunoreactivity (50.4 +/- 29.6, mean +/- standard deviation) that was not statistically different from that seen in SCCT (75.2 +/- 36.3). On the other hand, p53 immunostaining was virtually absent in TC cases (positivity for p53 was observed in only one instance in < 1% of cells). As for p27Kip1, the mean percentage of positive cells in PTT (82.7 +/- 9.9) was slightly lower than in TC (90.6 +/- 4.6) but significantly higher than in SCCT (53.4 +/- 30). CONCLUSIONS: The similar p53 immunoreactivity in both PTT and SCCT favour the interpretation of the former as carcinoma, albeit one whose behaviour would be tempered by the well-known regulatory effect exerted by p27Kip1 on the cell cycle. PMID- 11422484 TI - Host ethnicity influences non-Hodgkin's lymphoma subtype frequency and Epstein Barr virus association rate: the experience of a multi-ethnic patient population in Malaysia. AB - AIMS: The pattern of malignant lymphoma is known to vary in different populations. This study aims to elucidate the effect of ethnicity on subtype frequency of non-Hodgkin's lymphoma and EBV association rate. METHODS AND RESULTS: A total of 232 reconfirmed lymphoma cases in Malaysian patients were retrieved from the archives in the Department of Pathology, University Hospital, Kuala Lumpur. There were 24 (10%) Hodgkin's and 208 (90%) non-Hodgkin's lymphomas, 173 of the latter were in adult group (aged > or = 15 years). The ethnic composition were 41 Malays, 107 Chinese, 21 Indians and four none of the above. A male : female ratio of 2.4 : 1 was observed. Complete immunohistochemical studies in 158 cases revealed 36 (23%) T-cell, 121 (76%) B cell and one (1%) null-cell phenotype. Seventy-five percent of the T-cell lymphomas were peripheral T/NK-cell types. Among the classifiable lesions, low grade/indolent lymphomas constituted 17%: 2% were the lymphocytic subtype and 10% were follicular lymphomas. Approximately one-third of the follicular lymphomas occurred in Indian patients. The largest group of high-grade lymphoma was diffuse large B-cell type (46%), followed by peripheral T/NK-cell (18%). A predominance of NK/T-cell lymphomas occurred in Chinese (5/7), and all were EBV associated. Burkitt's lymphoma accounted for 5% (eight cases), all were Chinese males, with a 38% EBV-association rate. The frequency of EBV-associated B-cell lymphoma is three times more common in Chinese than Malays. The EBV positivity rate among lymphomas in ethnic Malay, Chinese and Indian patients was 5%, 15% and 22%, respectively, and in T- and B-cell lymphomas was 36% and 7%, respectively. CONCLUSIONS: This Malaysian series reveals differences in the subtype frequencies of non-Hodgkin's lymphomas and EBV association rate amongst patients of various ethnic groups residing in the same environment. PMID- 11422485 TI - Angiogenesis is redundant for tumour growth in lymph node metastases. AB - AIMS: Angiogenesis is essential for the growth of solid tumours. As the role of angiogenesis is unclear in the pathogenesis of primary lymph node (LN) tumours, we wondered whether neoangiogenesis was important in supporting and promoting the growth of tumours in LNs. METHODS AND RESULTS: We investigated 16 cases of squamous carcinoma involving oral cavity (n=9) and larynx/pyriform fossa (n=7), all of whom had lymph node metastases. Sections of the primary tumour, uninvolved mucosa, metastatic LN and nonmetastatic LN were double-immunostained with factor VIII-related antigen and MIB-1. Proliferating blood vessels, i.e. neoangiogenesis, was identified by coexpression of factor VIII-related antigen stained blood vessels and MIB-1 staining of the endothelial cell nuclei. Counts were performed in an area of 4 mm2. Primary tumours (127-188.7), uninvolved mucosa (54-84.5) and metastatic LNs (123.5-167) had significantly lower vessel counts than nonmetastatic LNs (194-253.9) (P=0.003; P < 0.001; P < 0.001, respectively). With regard to neoangiogenesis, primary tumours (1.7-5) had significantly higher counts than uninvolved mucosa (0-0.4), nonmetastatic LNs (0 0.4) and metastatic LNs (0.9-2.4) (P < 0.001; P < 0.001; P=0.047, respectively). CONCLUSIONS: Our data suggest that because of the rich native vascularity of lymph nodes, neoangiogenesis is redundant for the growth of metastatic tumour. PMID- 11422486 TI - Immunohistochemistry and the diagnosis of malignant mesothelioma. PMID- 11422488 TI - Immunohistochemical evidence for mesothelial origin of paratesticular adenomatoid tumour. PMID- 11422489 TI - Pericardial rhabdomyomatous spindle cell thymoma with mucinous cystic degeneration. PMID- 11422490 TI - Cell cycle-related proteins p21 and bcl-2: markers of differentiation in the human fallopian tube. PMID- 11422491 TI - Myofibrosarcoma of the nasal cavity and paranasal sinus. PMID- 11422492 TI - Clinical implications of telomerase detection. AB - In 1994 a sensitive method for the detection of telomerase was described. This assay, which was based on the polymerase chain reaction, suggested that telomerase activity was associated with immortal and cancer cells. Since then more than a thousand studies have documented the expression and activity of the enzyme in diseased tissues, primarily tumours. This review gives an overview of the biological significance of telomerase expression and methods for detecting its activity. This is followed by an organ system-based discussion of expression in normal tissues and disease states. We finish with speculation as to the future role of telomerase detection in diagnostic histopathology. PMID- 11422493 TI - Myofibroblastic differentiation in malignant fibrous histiocytoma (pleomorphic myofibrosarcoma): a clinicopathological study. AB - AIMS: We compared the clinical and pathological features of pleomorphic malignant fibrous histiocytoma (MFH)-like soft tissue sarcomas with and without myofibroblastic differentiation on electron microscopy. METHODS AND RESULTS: Fifty-three soft tissue tumours designated as MFH by light and electron microscopy were reassessed. Eighteen were specifically diagnosed and excluded, and follow-up (FU) information obtained for 24 of the other 35 cases. Myofibroblastic ultrastructure was seen in 7/24 (29%). Seventeen of 24 (71%) lacked myofibroblasts on electron microscopy, which showed fibroblastic or undifferentiated cells. Histologically, all tumours but one had storiform pleomorphic areas; one myofibroblastic neoplasm was fascicular and myxoid. No other morphological differences were seen. In seven myofibroblastic cases, smooth muscle in four cases and muscle-specific actin in two cases, desmin in three cases and S100 in one case were present. In 15 other tumours, smooth muscle in five cases and muscle-specific actin in one case, and desmin in one case were present; none of these cases expressed S100. CD34 was found in the myxoid areas of one myofibrosarcoma and 3/15 other tumours. Positivity for bcl-2 was seen only in non-myofibroblastic sarcomas (4/14). On follow-up (median 41 months), 2/7 (29%) myofibroblastic tumours recurred, 5/7 (71%) metastasized, and 3/7 (43%) patients died of disease. Among the non-myofibroblastic sarcomas, with a median follow-up of 47 months, 6/17 cases (35%) recurred, 10/17 (59%) metastasized, and 7/17 patients (41%) died of disease. CONCLUSIONS: Pleomorphic sarcomas with and without myofibroblastic differentiation on electron microscopy are clinically and histologically similar. The former display myoid immunohistochemical markers more frequently. PMID- 11422494 TI - Development of follicular dendritic cell sarcoma in hyaline-vascular Castleman's disease of the nasopharynx: tracing its evolution by sequential biopsies. AB - AIMS: Hyaline-vascular Castleman's disease (HVCD) and follicular dendritic cell (FDC) sarcoma occurring in the nasopharynx are both extremely rare. We report the first case of transformation of the former into the latter as documented by sequential biopsies. The steps involved in the transformation were described in detail and the possible role of p53 studied. METHODS AND RESULTS: The patient presented at the age of 23 years with nasopharyngeal HVCD. Hyaline- vascular Castleman's disease with FDC overgrowth was diagnosed in a recurrence 8 years later, and a frank FDC sarcoma developed at the same site 11 years after initial presentation. The patient remained disease-free 3 years after excision and adjuvant chemotherapy. The FDC sarcoma comprised swirling fascicles of spindly cells with indistinct cell borders. The tumour cells expressed the FDC markers CD21, CD35 and CNA.42 and in-situ hybridization for Epstein-Barr virus-encoded RNAs was negative. Over-expression of p53 protein was observed in the FDC sarcoma and an increased number of weakly p53-positive spindly cells could also be demonstrated in the HVCD specimen. This finding suggested a possible role of p53 in the evolution from HVCD to FDC sarcoma. Critical analysis of the literature shows that, among the 13 reported cases of FDC sarcoma associated with Castleman's disease, possible progression from the latter to the former is documented in only two cases. CONCLUSIONS: The sequential changes observed in the current case provide further evidence to strengthen the role of HVCD as a possible precursor of FDC sarcoma. There is a possible role of p53 in the transformation process but confirmation by future studies is needed. PMID- 11422495 TI - The spectrum of micronodular thymic epithelial tumours with lymphoid B-cell hyperplasia. AB - AIMS: A rare type of thymoma, micronodular thymoma with lymphoid B-cell hyperplasia, was recently reported by Suster and Moran. Thymic epithelial tumours with a similar pattern but with varied cytological features of the tumour cells are analysed. METHODS AND RESULTS: A total of 11 cases of thymic epithelial tumours characterized by micronodular proliferation of tumour cells separated by abundant lymphoid stroma with prominent germinal centres were reviewed clinicopathologically and examined immunohistochemically. The presence of Epstein Barr virus (EBV) genome was also examined by in-situ hybridization. Based on the morphology of tumour epithelial cells, cases were subdivided into four groups: group 1 (two cases) having spindle epithelial cells; group 2 (two cases) showing an admixture of spindle and polygonal epithelial cells; group 3 (five cases) having polygonal epithelial cells, with mild to moderate cytological atypia in four cases, and group 4 (two cases) representing lymphoepithelioma-like carcinoma. The degree of cytological atypia and the number of tumour cells positive for MIB-1 and p53 gradually increased towards group 4. The abundant lymphoid stroma in all cases contained many CD20-positive B-cells and CD3 and CD45RO-positive T-cells. CD99-positive immature T-cells were present in all cases of groups 1 and 2 and in most cases of group 3, but not in both cases of group 4 tumours. IgG, IgM and IgD-positive plasma cells and lymphocytes were also present in all cases, more prominent in those of groups 3 and 4. The EBV genome was detected in only a few lymphocytes in five cases. CONCLUSIONS: The tumours in this series belong to a distinct category of thymic epithelial tumours and each of the above groups may constitute a spectrum in the continuum of cytological atypia. The aetiological relationship of EBV with these tumours could not be proved. The lymphoid B-cell hyperplasia may result from a host immune response and may suggest a favourable clinical course of this type of tumour. PMID- 11422496 TI - Combined use of novel epithelial (MOC-31) and mesothelial (HBME-1) immunohistochemical markers for optimal first line diagnostic distinction between mesothelioma and metastatic carcinoma in pleura. AB - AIMS: To determine the value of immunohistochemistry in differentiation of malignant pleural mesothelioma from carcinoma in a pleural biopsy we optimized a double panel of MOC-31 and HBME-1 and compared the results with others from the literature. METHODS AND RESULTS: A multi-antibody panel was applied to biopsy samples from 44 cases of malignant pleural mesothelioma and 23 cases of carcinoma metastatic to the pleura. We used monoclonal antibodies against keratins, epithelial membrane antigen (EMA), epithelial antigen Ber-EP4, carcinoembryonic antigen (CEA), tumour-associated glycoprotein (B72.3), LeuM1, vimentin, desmin, epithelial related antigen (MOC-31) and mesothelial cell (HBME-1). Positivity for MOC-31 and Ber-EP4 was found to have the highest nosologic sensitivity (94.1% and 84.6%, respectively) and specificity (86.3% both antibodies) for carcinoma. Positive staining for HBME-1 and vimentin had the highest sensitivity (90.9% and 100%, respectively) and specificity (91.3% and 60%, respectively) for mesothelioma. A two-marker antibody panel with HBME-1 and MOC-31 was the most efficient for the distinction between carcinoma and malignant pleural mesothelioma. CONCLUSION: A combination of MOC-31 (an anti- epithelial marker) and HBME-1 (an anti-mesothelial marker) has a diagnostic efficiency of 76.1% for the distinction between carcinoma and mesothelioma in pleura. PMID- 11422497 TI - p27 immunostaining is related to prognosis in malignant mesothelioma. AB - AIMS: To evaluate the prognostic value of the proliferation-associated antigen p27 in malignant mesothelioma. METHODS AND RESULTS: Sections from 36 patients with malignant mesothelioma were immunohistochemically stained for the p27 antigen. The results were quantified by recording the proportion of positively stained nuclei and these data were tested for association with patient survival. p27 expression was also compared with tumour type, MIB-1 expression, apoptotic and mitotic indices. Patients with pleural malignant mesothelioma showing low p27 expression (< 53% of cells positive) survived for significantly shorter periods of time than those with high p27 expression (> or = 53% of cells positive) (P=0.04). The 50% survival time of subjects with low p27 was 4-6 months whereas for those with high p27 expression it was 10-11 months. There was no significant association between p27 expression and MIB-1, apoptotic or mitotic indices. CONCLUSIONS: Low p27 expression in pleural malignant mesothelioma is associated with a significantly worse prognosis. p27 appears to be an independent prognostic variable which may be of value in counselling patients with malignant mesothelioma. PMID- 11422498 TI - Monoclonal antibody SM047 as an immunohistochemical marker of ovarian adenocarcinoma. AB - AIMS: This study describes the generation of a monoclonal antibody designated SM047 which binds to an epitope that is displayed by a multivalent antigen associated with the glycocalyx of ovarian adenocarcinoma cells. The study also investigates SM047 staining in adenocarcinomas of diverse sites in order to determine whether the antibody is specific for ovarian adenocarcinoma and of value in the confirmation of an ovarian origin when the site of primary tumour is unknown. METHODS AND RESULTS: SM047, an IgM monoclonal antibody, was the product of hybridoma cells derived from fusion of SP2 myeloma cells with splenocytes of a mouse that had been immunized with a membrane preparation of tumour (ovarian serous cystadenocarcinoma) and boosted with cells from a cell line established from a similar tumour in a different patient. Sixty-two primary ovarian adenocarcinomas (28 serous, 23 mucinous, five endometrioid and six clear cell), 69 adenocarcinomas arising primary at other sites and 10 mesotheliomas were stained with SM047. There was positive membrane staining, which was usually strong and widespread, in 27 of 28 ovarian serous carcinomas and in all ovarian endometrioid and clear cell carcinomas. Most ovarian mucinous tumours were negative or exhibited weak cytoplasmic staining. Staining was variable in the other tumours but there was positive staining of most endometrial, endocervical and pancreatic adenocarcinomas. Most colonic adenocarcinomas were negative or exhibited weak cytoplasmic staining. CONCLUSIONS: SM047 is strongly expressed in most ovarian serous adenocarcinomas and in other female genital tract adenocarcinomas, with the exception of ovarian mucinous tumours. The antibody may be useful in confirming the ovarian origin of an adenocarcinoma when used as part of a larger panel. This is especially so in the distinction between a non mucinous ovarian adenocarcinoma, which usually exhibits strong membranous staining, and a colonic adenocarcinoma which is usually negative or exhibits weak cytoplasmic staining. These findings need to be confirmed by further study of larger numbers of cases. PMID- 11422499 TI - Expression of MUC1 and MUC2 and carbohydrate antigen Tn change during malignant transformation of biliary papillomatosis. AB - AIM: Biliary papillomatosis is characterized by papillary proliferations of biliary lining cells without invasion or metastasis. The neoplastic character and biological behaviour of this disease remain still speculative. These issues were examined in this study. METHODS AND RESULTS: Mucin core protein MUC1, MUC2, MUC3, MUC5AC and carbohydrate antigens (T, Tn and sialosyl Tn) were immunohistochemically examined, using 11 lesions of biliary papillomatosis from seven patients, and five lesions of biliary papillomatosis with foci of carcinoma from four patients. Five cases of papillary intrahepatic cholangiocarcinoma and 12 histologically normal livers were used as a control. Patients with biliary papillomatosis alone or with carcinoma were middle-aged or elderly (five men and six women). Microscopically, biliary papillomatosis showed a villous, papillo tubular, papillary, or papillo-villous pattern with a thin fibrovascular core. Cytologically, they were classifiable into biliary epithelial or pyloric gland like type. The former was frequent in the cases associated with carcinoma. Expression of MUC1, Tn antigen and sialosyl Tn antigen was frequent and marked in biliary papillomatosis alone and with carcinoma and also intrahepatic papillary carcinoma. In addition, marked expression of MUC1 and Tn antigen were rather frequent in biliary papillomatosis with carcinoma and intrahepatic biliary papillary carcinoma compared with biliary papillomatosis. MUC2 was rather frequent and marked in biliary papillomatosis alone compared to other two disease groups. Focal expression of MUC5AC and MUC2 was rather frequent and infrequent irrespective of disease group, respectively. Focal expression of T antigen was frequent in papillary ICC. CONCLUSION: Biliary papillomatosis could undergo overt malignant transformation along with altered phenotypic expression of MUC proteins and mucin carbohydrate antigens. PMID- 11422500 TI - Histological changes in the hypofunctional pituitary gland following conventional radiotherapy for adenoma. AB - AIMS: Although delayed hypopituitarism is a common complication of conventional radiotherapy of sellar tumours, histological changes that may account for it have been rarely reported. To elucidate the changes, hypofunctional pituitary glands following irradiation were studied. METHODS AND RESULTS: Two pituitary glands obtained at autopsy from patients who had been irradiated for adenoma and exhibited hypopituitarism were examined. In both cases diffuse fibrosis was observed in the adenohypophysis, whereas the neurohypophysis remained unchanged. Immunohistochemistry showed that stellate-shaped S100 protein-positive cells were increased in number and distributed among the endocrine cells. Some irradiated endocrine cells showed dense granular immunoreactivity for mitochondrial protein, cytochrome oxidase and manganese-superoxide dismutase. In addition to faint reactivity with anti-cytokeratin 8, 18 antibody, many cells were densely positive with anti-cytokeratin 1, 5, 10, 14 antibody. CONCLUSIONS: These results indicate that radiation- induced fibrosis is associated with an increased number of folliculo-stellate cells and the presence of metabolic dysfunctional mitochondria resembling mitochondria in oncocytes. Squamous metaplasia in the irradiated endocrine cells was also noted. Various intracellular changes may participate in delayed pituitary hypofunction following radiotherapy. PMID- 11422501 TI - Increased microvessel density in malignant and borderline mammary phyllodes tumours. AB - AIMS: Tumour vascularity is considered a prognostic indicator in breast carcinoma, but its utility in mammary phyllodes tumour has not been explored. The authors report the correlation between intratumoral microvessel density and the histological grade of phyllodes tumour. METHODS AND RESULTS: Forty cases of phyllodes tumour were reviewed for stromal cellularity, overgrowth, cytological pleomorphism, mitotic count and margin pattern. Using established criteria, these were diagnosed as benign (n=28), borderline (n=10) and malignant (n=2). Microvessel density was counted on CD31-stained slides as the number of vessels per high power field. For benign phyllodes tumour, the range was 7-26.2 (mean 13.1); for borderline phyllodes tumour the range was 17.2-32.5 (mean 22.4); for malignant phyllodes tumour the range was 25.9-33.3 (mean 29.6). The difference between the benign and borderline groups was significant (P < 0.0001) but that between the borderline and malignant groups was not, due to the small number of malignant cases. CONCLUSIONS: There is a significant difference in stromal microvessel density between benign and borderline phyllodes tumour. Although the small number of cases of malignant phyllodes tumour limits further interpretation, we believe that microvessel density can be used as an additional objective histological parameter in the evaluation of phyllodes tumour. PMID- 11422502 TI - Histomorphometric assessment of bone turnover in uraemic patients: comparison between activation frequency and bone formation rate. AB - AIMS: The histomorphometric assessment of bone formation rate (BFR/BS) in bone biopsies from uraemic patients is of crucial importance in differentiating low from high turnover types of renal osteodystrophy. However, since BFR/BS relies on osteoblasts, activation frequency (Ac.f), encompassing all remodelling phases, has recently been preferred to BFR/BS. This study was carried out to consider whether estimation of Ac.f is superior, in practical terms, to that of BFR/BS in distinguishing between different rates of bone turnover in uraemic patients. METHODS AND RESULTS: Bone biopsies from 27 patients in predialysis (20 men and seven women; mean age 53 +/- 12 years) and 37 in haemodialysis (22 men and 15 women; mean age 53 +/- 12 years) were examined. The types of renal osteodystrophy were classified on the basis of morphology. Bone formation rate and Ac.f were evaluated according to standardized procedures. The Ac.f was calculated both as a ratio between BFR/BS and wall thickness (W.Th) and as a reciprocal of erosion, formation and quiescent periods (EP, FP and QP). Patients were affected by renal osteodystrophy with predominant hyperparathyroidism (two predialysis and 16 dialysis), predominant osteomalacia (three predialysis and seven dialysis) or that of advanced (nine predialysis and five dialysis) or mild (seven predialysis and four dialysis) mixed type or adynamic type (six predialysis and five dialysis). Activation frequency, which with either formula requires the measurement of W.Th, i.e. the thickness of bone structural units (BSUs), was not calculated in three dialysis patients with severe hyperparathyroidism and in one predialysis and four dialysis patients with severe osteomalacia, because only incomplete BSUs were found. In dialysis, EP was higher in the adynamic than in the other types of osteodystrophy. During both predialysis and dialysis, FP was higher in osteomalacia than in the other forms of osteodystrophy, and in adynamic osteopathy than in hyperparathyroidism or in advanced and mild mixed osteodystrophy. During predialysis and dialysis, QP was higher in the adynamic than in the other forms of osteodystrophy. Correlations were found between BFR/BS and Ac.f, during predialysis (r=0.97) and dialysis (r=0.95). CONCLUSIONS: The superiority of Ac.f in assessing bone turnover, in comparison to BFR/BS, is conceptual rather than practical. The highest values for FP in osteomalacia and for QP in adynamic bone allow a clearer characterization of these low turnover conditions. PMID- 11422503 TI - Portals and meta-indices. PMID- 11422504 TI - Squamous cell carcinoma in a giant oesophageal fibrovascular polyp. PMID- 11422505 TI - Spindle cell haemangioendothelioma and germinoma of the mediastinum. PMID- 11422506 TI - Genetic transformation of the housefly Musca domestica with the lepidopteran derived transposon piggyBac. AB - The piggyBac transposable element was successfully used for stable genetic transformation of the housefly Musca domestica. The construct contains the EGFP marker under the control of Pax-6 binding sites, which can drive eye-specific expression in insect species as distantly related as Drosophila melanogaster and Tribolium castaneum [Berghammer, A.J., Klingler, M. and Wimmer, E.A. (1999) Nature 402: 370-371]. We obtained seven independent integration events among 41 fertile G0 Musca flies. Most of the transformed lines contained two or more chromosomal insertions of the EGFP marker which were stably inherited over more than 15 generations. piggyBac-mediated transposition was verified by identifying the characteristic TTAA duplication at the insertion sites. This first report of stable transmission of a genetic marker in Musca confirms the use of this vector marker system for effective gene transfer in a broad range of insect species. PMID- 11422507 TI - Molecular characterization of a peroxiredoxin from the hard tick Haemaphysalis longicornis. AB - Antioxidant enzymes in eukaryotes play an important role in protection against the oxygen radicals generated during aerobic metabolism. Here we report the cloning and characterization of a cDNA encoding the antioxidant enzyme peroxiredoxin from the hard tick Haemaphysalis longicornis (HlPrx). HlPrx is 939 bp long and contains a 101 bp non-translated sequence at the 5' end and a polyadenylation singnal followed by a poly(A) tail at the 3' end. HlPrx encodes a full-length protein with a predicted molecular mass of 26 kDa that possesses one cysteine residue at amino acid 49 that is conserved among Prx proteins of various species. GenBanktrade mark analysis showed that the deduced amino acid sequence had significant similarity to mammalian and plant Prxs at the amino acid level. A DNA-nicking assay revealed that Escherichia coli-expressed recombinant HlPrx (rHlPrx) inhibited oxidative-nicking of supercoiled plasmid DNA. Two-dimensional immunoblot analysis with mouse antirHlPrx serum showed reaction with a major constituent protein spot in extracts of adult ticks. In addition, immunoblot analysis showed that rHlPrx was immunoreacted with serum from rabbits repeatedly infested with H. longicornis. Localization analysis using mouse antirHlPrx serum revealed that native HlPrx was highly expressed in the salivary gland of the tick. Moreover, Northern blot analysis showed that the level of HlPrx transcripts was increased during blood sucking. The present results indicate that HlPrx may be an important detoxifying enzyme during the normal life span as well as during blood sucking in ticks. PMID- 11422508 TI - Cloning of two novel P450 cDNAs from German cockroaches, Blattella germanica (L.): CYP6K1 and CYP6J1. AB - Two novel P450 cDNAs, CYP6K1 and CYP6J1, were isolated from German cockroaches, Blattella germanica (L). Both CYP6K1 and CYP6J1 are typical microsomal P450s and their deduced amino acid sequences share a number of common characteristics with other members of the P450 superfamily. Both CYP6K1 and CYP6J1 showed the highest per cent identity (based on the deduced amino acid sequence) to CYP6L1 from B. germanica and CYP6H1, a putative ecdysone 20-hydroxylase from Locusta migratoria. Using a CYP6K1 probe, two mRNA signals (~2.5 and ~2.1 kb) were detected in all life stages. Both signals were just detectable in the eggs and became stronger in later instars. The strongest signals were detected in the fifth and sixth instars as well as in adults. These two bands were also detected in the abdomens and in the remainder of bodies of both male and female adults. Southern blots suggest the two mRNA bands detected in the Northern blot might be a result of alternative splicing. No signal could be detected at any life stage using the CYP6J1 probe, suggesting that CYP6J1 was expressed at a low level. PMID- 11422509 TI - Expression and induction of three family 4 cytochrome P450 (CYP4)* genes identified from insecticide-resistant and susceptible western corn rootworms, Diabrotica virgifera virgifera. AB - We have previously determined that cytochrome P450-based oxidation is involved in resistance to the insecticides methyl parathion and carbaryl in geographically distinct Nebraska western corn rootworm populations. Three new family 4 cytochrome P450 (CYP4) gene fragments (CYP4AJ1, CYP4G18 and CYP4AK1) were cloned and sequenced from insecticide-resistant and -susceptible western corn rootworms. Insecticide bioassays indicated the resistant population employed in this study was significantly resistant to the insecticides methyl parathion and carbaryl. CYP4AJ1 and CYP4G18 were cloned from both genomic PCR and RT-PCR products, although only CYP4AJ1 contains an intronic region. Alignments of inferred amino acid sequences with other homologous insect CYP4 genes indicates a high degree of similarity. Northern analysis concurrently employing mixed probes representing each of the three rootworm CYP4 fragments identified increased mRNA transcript signals (i) in resistant rootworms and (ii) following induction by the P450 inducer pentamethyl benzene. These results support our previous documentation of P450-based insecticide resistance and suggest increased CYP4 transcript abundance can serve as a molecular resistance-associated marker. PMID- 11422510 TI - The juvenile hormone binding protein of silkworm haemolymph: gene and functional analysis. AB - A cDNA fragment of haemolymph juvenile hormone binding protein (hJHBP) from larvae of Bombyx mori was amplified by RT-PCR using degenerate primers based on the N-terminal amino acid sequence of purified hJHBP and a conserved region near the C-terminus of other lepidopteran hJHBPs. 5'- and 3'-ends were amplified by RACE to yield cDNAs, hJHBP1 and hJHBP2, encoding 225 amino acids with three substitutions. hJHBP-mRNA levels in the fat body were constant in the 4th instar, but decreased in the 5th. JHBP protein was constant until wandering, then declined. Recombinant hJHBP1 expressed in E. coli migrated on SDS-PAGE with a Mr of 32 kDa and showed a Kd of 4.5 x 10-7 M with JH III, both similar to those of native hJHBP. PMID- 11422511 TI - Hermes-mediated germ-line transformation of the Mediterranean fruit fly Ceratitis capitata. AB - We report the use of the Hermes transposable element for germ-line transformation of the Mediterranean fruit fly, Ceratitis capitata. Hermes was able to genetically transform this insect at an estimated frequency between 0.6 and 1.1%, which is comparable to the transformation frequencies obtained for this species when using other transposable elements. Hermes integrates into the medfly genome by a cut-and-paste mechanism and the sequences integrated into the genome are delimited by the terminal nucleotides of the Hermes inverted terminal repeats. Integration resulted in the generation of 8 bp target site duplications, the sequences of which conformed to the target site duplications generated by hAT element transposition in insects. The Hermes element is one additional genetic tool that can be deployed in manipulating and characterizing the medfly genome. PMID- 11422512 TI - Wolbachia in the Asian rice gall midge, Orseolia oryzae (Wood-Mason): correlation between host mitotypes and infection status. AB - Using a PCR-based method, we detected Wolbachia in the Asian rice gall midge. Furthermore, results showed that all females across all biotypes are infected with Wolbachia. However, all male flies are not infected and show different infection frequency in different biotypes. We have also identified three mitotypes, in the rice gall midge, based on DraI restriction pattern of a portion of the 12S rRNA gene that was PCR amplified using primers specific to this gene. All the females and infected male flies had type 1 mtDNA while uninfected males showed only type 2 or 3 mtDNA. Inheritance patterns of mtDNA revealed the existence of a correlation between mtDNA type and Wolbachia infection in the Asian rice gall midge. Evidence for paternal inheritance of mtDNA in Wolbachia free gall midge is also presented. PMID- 11422513 TI - Cloning of a catalytic subunit of cAMP-dependent protein kinase from the honeybee (Apis mellifera) and its localization in the brain. AB - In the honeybee the cAMP-dependent signal transduction cascade has been implicated in processes underlying learning and memory. The cAMP-dependent protein kinase (PKA) is the major mediator of cAMP action. To characterize the PKA system in the honeybee brain we cloned a homologue of a PKA catalytic subunit from the honeybee. The deduced amino acid sequence shows 80-94% identity with catalytic subunits of PKA from Drosophila melanogaster, Aplysia californica and mammals. The corresponding gene is predominantly expressed in the mushroom bodies, a structure that is involved in learning and memory processes. However, expression can also be found in the antennal and optic lobes. The level of expression varies within all three neuropiles. PMID- 11422514 TI - cDNA cloning and transcript distribution of two different neuroparsin precursors in the desert locust, Schistocerca gregaria. AB - Neuroparsins were originally identified in locust corpus cardiacum extracts as folliculostatic or 'antigonadotropic' neuropeptides. This paper presents the cloning of two different neuroparsin precursor cDNAs from the brain of the desert locust, Schistocerca gregaria. The first transcript encodes the precursor (Scg NPP1) of S. gregaria neuroparsin A and B, whereas the second codes for a novel neuroparsin-related peptide precursor (Scg-NPP2). Both precursors display significant sequence similarities with each other and with the Locusta migratoria neuroparsin (Lom-NPP) and Aedes aegypti ovary ecdysteroidogenic hormone (Aea OEH1) precursors. Northern blot analysis revealed that these neuroparsin transcripts are present in larval and adult locust brains. Interestingly, the Scg NPP2 mRNA content proved to be strongly regulated during the reproductive cycle in both adult males and females. PMID- 11422515 TI - Pemphigus and diet: does a link exist? PMID- 11422516 TI - Unusual manifestations of dermatophytoses. PMID- 11422517 TI - Dermatoepidemiology: wherefore art thou in this perilous time of need? PMID- 11422518 TI - Descriptive epidemiology of skin cancer on Aruba: 1980-1995. AB - BACKGROUND: An epidemiology unit was established in Aruba in 1994. The primary focus was the development of an infrastructure for both surveillance and health information data systems. This effort resulted in the first analysis of available skin cancer data. METHODS: A retrospective study of cancer and particularly skin cancer cases at the local histopathology laboratory for the period 1980-1995 was performed. All available records with a cancer diagnosis were evaluated. RESULTS: During the study period, an increase in age-adjusted rates for both men and women was observed for basal cell carcinomas and squamous cell carcinomas. No temporal trend was discerned for malignant melanoma or other skin cancers because the numbers of cases were small. Lesions were most often seen on the nose, face, and arm. Dermatologists provided the majority of skin cancer diagnoses and demonstrated the greatest diagnostic accuracy. CONCLUSIONS: The presence of a central histopathology laboratory may provide small island states, like Aruba, with important public health data. This first description of Aruba's skin cancer epidemiology provides information that may be utilized for future public health action. PMID- 11422519 TI - Type III Ehlers-Danlos syndrome: correlations among clinical signs, ultrasound, and histologic findings in a study of 35 cases. AB - BACKGROUND: Patients with type III Ehlers-Danlos syndrome show hypermobile joints, luxation, and minimal atrophic scars. The disease has an incidence of 9 16% and the molecular defect that underlies this subtype is unknown. In order to widen diagnostic investigations in type III Ehlers-Danlos syndrome, skin and articular areas were studied by ultrasound (US) test. METHODS: Thirty-five patients, 20 women and 15 men, aged from 18 to 25 years, with type III Ehlers Danlos syndrome were enrolled in this study. Patients showed hypermobile joints (35 cases), minimal atrophic wrinkled scars (35 cases), and a few ice-pick acne scars. Hypermobility of phalangeal joints was studied by means of the Beighton scoring system and by a US test (performed by Sonora Logic 400 MD unit with 10 MHz probe). The following US parameters were considered: the distance of the intra-articular space and the thicknesses of the extensor tendon and of the overhanging dermis and subcutis. RESULT: US reports showed an increase in distance of the intra-articular space and a reduction in the thickness of the extensor tendon and of the dermis and subcutis covering it. The dermis showed high and homogeneous echogenicity with irregular hyperechogenic lines. CONCLUSION: The Beighton scoring system, the US findings, and the aspect and diameter of the scars suggested that the disease can be divided into three different stages which allow the correlation, over progressive intervals of values, of the articular hypermobility and atrophic wrinkled scars. Our results showed that US test is able to confirm the clinical diagnosis and to evaluate the seriousness of hypermobile joints. PMID- 11422520 TI - Post-kala-azar dermal leishmaniasis in Nepal. AB - BACKGROUND: Post-kala-azar dermal leishmaniasis (PKDL) manifests as a skin eruption after healing of visceral leishmaniasis (VL), either spontaneously or as a result of treatment. This study was undertaken to describe the demographic, clinical, and histopathologic features of PKDL in Nepal. METHODS: Demographic, clinical, microbiologic, and histopathologic features and response to treatment were studied in 22 patients with PKDL from April 1998 to March 2000. RESULTS: PKDL accounted for 0.13% of all new dermatologic cases. There were 13 (59.1%) males and nine (40.9%) females. A past history of kala-azar was present in all but one patient. A family history of kala-azar was noted in eight (36.4%) patients. All patients presented with multiple types of lesion, except for two in whom only macular lesions were seen. Oral lesions in the form of nodules and plaques were seen in four patients. Generalized lymphadenopathy was present in five patients. Slit skin smears revealed Leishman-Donovan bodies (LDBs) in nine (40.9%) patients. In macular lesions, there was a sparse infiltrate of plasma cells, lymphocytes, or histiocytes in the upper dermis. There was a dense chronic inflammatory infiltrate comprising plasma cells, lymphocytes, histiocytes, and epithelioid cells in the entire dermis from papules, plaques, or nodules. Giemsa staining of biopsy specimens revealed LDBs in seven (38.9%) patients only. Fine needle aspiration from epitrochlear lymph nodes in two patients demonstrated LDBs. All patients responded well to treatment with minimal side-effects. CONCLUSIONS: This study emphasizes the need to be aware of the possibility of cases of PKDL in endemic regions of leprosy, as the conditions may be difficult to distinguish clinically and histopathologically. PMID- 11422521 TI - TNFalpha and IL-6 are mediators in the blistering process of pemphigus. AB - BACKGROUND: Pemphigus is an autoimmune disease characterized by intraepidermal blisters induced by pemphigus IgG. In addition to autoantibodies, molecular mechanisms involved in acantholysis remain largely unknown. For this reason, we address a possible role of the inflammatory cytokines IL-6 and TNFalpha in pemphigus lesions. METHODS: Sixteen biopsies from patients with different types of pemphigus were studied by in situ hybridization using DNA fluorescent probes for IL-6 and TNFalpha mRNA. RESULTS: Fifty-six percent of lesional biopsies exhibited cytokine gene expression, which was poorly expressed in noninvolved skin. Deposits of TNFalpha and IL-6 were products of in situ transcription at the epidermal level. CONCLUSIONS: Inflammatory cytokine expression around the blister could play a mediator role in pemphigus lesions by increasing epithelial damage. PMID- 11422522 TI - Neurologic variant of epidermal nevus syndrome with a facial lipoma. AB - A 1-year-old girl presented for evaluation of a linear plaque on her forehead. She was born at 36 weeks' gestation following an uncomplicated pregnancy and delivery. At birth, she was noted to have an enlarged right cheek. She had no seizure history, but developed grand mal seizures 1 year later. On examination, she had a yellow plaque on her forehead which extended onto her nose. Under her right jaw, extending onto her anterior neck, there was a cafe-au-lait macule within which there was a yellow plaque which followed the lines of Blaschko. Her right cheek was enlarged and was erythematous (Fig. 1). Magnetic resonance imaging (MRI) of her face showed a mass in the right cheek deep to the subcutaneous fat tissue layer. The signal from the mass was identical to that from the fat, indicating that the mass represented a lipoma. This was later excised surgically and was histologically a lipoma. MRI of the brain demonstrated enlargement of the right lateral ventricle in addition to enlargement of the right cerebral hemisphere. There was also evidence of abnormal gyral architecture. Computerized tomography (CT) three-dimensional reconstruction of the skull demonstrated overgrowth of the right maxilla, right mandible, and right orbit (Fig. 2). PMID- 11422523 TI - Two cases of reactive perforating collagenosis arising at the site of healed herpes zoster. PMID- 11422524 TI - Erythema induratum with pulmonary tuberculosis: histopathologic features resembling true vasculitis. PMID- 11422525 TI - Primary cutaneous nocardiosis associated with intra-articular corticosteroid injection. PMID- 11422526 TI - Early congenital syphilis. PMID- 11422527 TI - Rosacea fulminans (pyoderma faciale): successful treatment of a 3-year-old girl with oral isotretinoin. PMID- 11422528 TI - Re-infection (secondary) inoculation cutaneous tuberculosis. PMID- 11422529 TI - Investigator-masked comparison of tazarotene gel q.d. plus mometasone furoate cream q.d. vs. mometasone furoate cream b.i.d. in the treatment of plaque psoriasis. PMID- 11422530 TI - Melasma treated with hydroquinone, tretinoin, and a fluorinated steroid. PMID- 11422531 TI - Anthrax: an overview within the Indian subcontinent. PMID- 11422532 TI - Basis of occlusive therapy in psoriasis: correcting defects in permeability barrier and calcium gradient. AB - BACKGROUND: Although occlusive dressings have great potential in the management of psoriasis vulgaris, the therapeutic mechanism is not completely understood. Occlusion artificially restores and corrects the defective barrier in psoriasis plaques. Additionally, occlusion is know to normalize the epidermal calcium gradients in hyperproliferative murine skin models. METHODS: To investigate the basis of the therapeutic effect of occlusion on psoriatic plaques, we investigated the ultrastructural morphology of intercorneocyte lipid layers, lamellar bodies, and calcium gradient in chronic plaque-type psoriasis after occlusion with a water vapor-impermeable membrane. The specimens were processed for electron microscopy using: (i) ruthenium tetroxide postfixation; and (ii) ion capture cytochemistry for calcium localization. RESULTS: Occlusion for 7 days resulted in a nearly mature pattern of intercellular multilamellar structures, re establishment of the near-normal epidermal calcium gradient, and disappearance of calcium precipitates from the stratum corneum interstices. CONCLUSIONS: The normalization of the permeability barrier and epidermal calcium gradient may play important roles in the therapeutic effects of occlusive dressings in chronic plaque-type psoriasis. PMID- 11422533 TI - Azone enhances clinical effectiveness of an optimized formulation of triamcinolone acetonide in atopic dermatitis. AB - BACKGROUND: Atopic dermatitis is a chronic, relapsing condition affecting up to 14% of the population in Western countries. Topical corticosteroids are the mainstay of treatment. Triamcinolone acetonide, a corticoid of intermediate potency, has proven useful in the treatment of atopic dermatitis. AIM: To evaluate the effectiveness of a triamcinolone acetonide-laurocapram combination in the treatment of atopic dermatitis. METHODS: One hundred and fifty patients were enrolled in a three-arm, parallel group, controlled clinical trial evaluating the effectiveness of a triamcinolone acetonide (0.05%) and laurocapram combination, applied twice daily for 2 weeks, in the treatment of atopic dermatitis. Fifty patients received triamcinolone acetonide-laurocapram (TNX), 50 triamcinolone acetonide (TN), and 50 a vehicle control formulation (AN). Response to treatment was evaluated by change in disease severity at 6 h, at 3, 8, and 15 days after the start of treatment, and by the global change in disease status. RESULTS: TNX effected a significantly higher degree of improvement in the signs and symptoms of atopic dermatitis (erythema, induration, and pruritus) and a greater overall improvement in disease status compared with treatment with TN or AN. Treatment-associated side-effects were local reactions, occurring in three, two, and six patients in the TNX, TN, and AN groups, respectively. CONCLUSIONS: The results suggest that the incorporation of laurocapram in the formulation enhances the effectiveness of triamcinolone acetonide, without compromising its safety profile. PMID- 11422534 TI - Head lice infestations in different ethnic groups. PMID- 11422535 TI - Chronic phlebopathic cutaneous ulcer: a therapeutic proposal. PMID- 11422536 TI - Viruses and cancer. PMID- 11422537 TI - Strategies in subversion: de-regulation of the mammalian cell cycle by viral gene products. PMID- 11422539 TI - Experimental observations in the rat on the influence of cadmium on skin wound repair. AB - Wound healing in the skin depends upon the availability of appropriate trace metals as enzyme cofactors and structural components in tissue repair. The present study forms part of a series of experimental investigations to examine the influence of xenobiotic elements with no known nutritional function and which are known to compete with essential trace metals. It was designed to investigate further the importance of trace metals in wound healing as an aid to wound management and to identify mechanisms of nonhealing which constitute a major problem in human medicine. Surgically induced skin wounds in young adult male Wistar rats were exposed topically to 0.2 ml of 0.01, 0.10 or 1.0% cadmium chloride (aq.) daily for up to 10 days. Control wounds received de-ionized water only. Wounds exposed to cadmium chloride at 0.01 or 0.10% healed in a similar fashion to controls and exhibited a comparable histological profile with metallothionein distribution. Wounds receiving 1.0% cadmium chloride failed to heal or fully re-epithelialize within 7 days and animals were humanely killed. They showed a persistent mass of inflammatory cell infiltration, oedema, wound debris and aberrant epidermal cell growth. Metallothionein concentrations in the epidermis and fibroblasts of the papillary dermis increased greatly by 5 days postwounding and remained high through the observation period. Cadmium was identified in the liver, kidney and wound sites. In the wound, 1.0% cadmium chloride induced statistically significant (P > 0.001) changes in local concentrations of zinc and calcium at key stages in the healing process, and as a consequence disturbed the trace metal balance necessary for normal wound repair. Zinc levels were increased twofold after 7 days, but calcium was markedly reduced. Local changes in the distribution of metallothionein indicate interaction of cadmium and trace metal carrier proteins as a probable mechanism for impaired wound healing. The cytotoxicity of cadmium is considered to be largely responsible. PMID- 11422538 TI - Biology of human papillomaviruses. AB - Human papillomaviruses (HPVs) cause squamous cancers of epithelial surfaces, of which genital cancers are the most common. In this article we have attempted to describe the properties and functions of the viral proteins of HPV type 16, a common cause of genital cancers, and have tried to suggest how their expression may lead to a dysregulated cell which may become malignant. These viruses are attempting to replicate in terminally differentiating keratinocytes and must stimulate G1 to S-phase progression for the replication of their genome. As part of the successful completion of replication and assembly of infectious virus particles, the virus needs at least partial differentiation to occur. Therefore, at the same time as differentiation is occurring, the nuclei of infected cells are in S-phase. While the mechanisms of action of the viral proteins are not completely understood, researchers are making progress and this article strives to bring together the conclusions from some of this work. PMID- 11422540 TI - Intravenous 1alpha, 25[OH]2 vitamin D3 (calcitriol) pulse therapy for bone lesions in a murine model of chronic cadmium toxicosis. AB - The aim of the present study was to clarify the therapeutic effects of 1alpha, 25[OH]2 vitamin D3 (calcitriol) pulse injection on bone lesions induced in a rat model of chronic cadmium toxicosis. Ovariectomized (OVX) and control-operated (sham-OVX) rats were given repeated intravenous injections of 0.5 mg/kg/day CdCl2 for 70 weeks. The rats were then treated intravenously with 0.02 microg/kg/day calcitriol 3 days per week for 8 weeks. CdCl2 treatment induced increases in osteoid volumes of the femur cortex and trabecula. This change was accompanied by an increase in the volume of iron deposition at the mineralization front of the trabeculae and a reduction in mineral density. Abnormalities of bone metabolic parameters, which were increases in the blood calcium, inorganic phosphorous, bone-specific alkaline phosphatase, parathyroid hormone (PTH) and osteocalcin levels, and in the urine deoxypyridinoline (D-PYR) level, were also induced. Calcitriol treatment increased the blood calcium and inorganic phosphorous levels, and reduced the blood PTH level. Decreases in blood tartrate-resistant acid phosphatase and urine d-PYR levels were also induced indicating that bone resorption was suppressed. The findings indicated that the increased osteoid volume of the cortex and Fe-deposition volume of the trabecula were improved. These effects or improvements were observed in the sham-OVX rats but not in the OVX rats. PMID- 11422541 TI - Skeletal muscle necrosis and regeneration after injection of Thalassophryne nattereri (niquim) fish venom in mice. AB - Stings by Thalassophryne nattereri are responsible for envenomation of fishermen in north-eastern Brazil. Its venom induces prominent local tissue damage, characterized by pain, oedema and necrosis. The pathogenesis of acute muscle damage induced by T. nattereri venom was studied in mice. Intramuscular injection induced myonecrosis within the first hours. Some muscle cells presented a hypercontracted morphology, but most necrotic fibres were not hypercontracted, being instead characterized by a disorganization of myofibrils, with Z line loss, mitochondrial swelling and sarcolemmal disruption. In addition, thrombosis was observed histologically in venules and veins, together with vascular congestion and stasis, evidenced by intravital microscopy. Venom induced a rapid increment in serum creatine kinase (CK) levels, concomitant with a reduction in gastrocnemius muscle CK activity, whereas no increments in muscle lactic acid were detected. A rapid cytolytic effect was induced by the venom on C2C12 murine myoblasts in culture. The inflammatory reaction in affected muscle was characterized by oedema and scarce cellular infiltrate of polymorphonuclear leucocytes and macrophages, with a consequent delay in the removal of necrotic material. Skeletal muscle regeneration was partially impaired, as evidenced by the presence of regenerating fibres of variable size and by the increase of fibrotic tissue in endomysium and perimysium. It is suggested that T. nattereri venom affects muscle fibres by a direct cytotoxic effect, and that the vascular alterations described preclude a successful regenerative process. PMID- 11422542 TI - The adrenaline rush: nursing students' experiences with the Northern Ireland Ambulance Service. AB - AIM: The aim of this project was to explore nursing students' experiences whilst on placement with the Northern Ireland Ambulance Service. BACKGROUND: The literature suggests that experiential learning can enhance understanding, knowledge and skills and has the potential for reducing the theory-practice gap. DESIGN: A qualitative approach was utilized, employing diaries and focus group interviews to gather data. One hundred and fifty-four nursing students submitted diaries, producing a response rate of 69%, while 190 students partook in interviews (86%). FINDINGS: Content analysis indicated that the students gained a new appreciation of the concept of holistic care, and that increased interprofessional understanding provides the foundation for improved quality of patient care. PMID- 11422543 TI - Beneath the surface: collaboration in alcohol and other drug treatment. An analysis using Foucault's three modes of objectification. AB - AIM OF STUDY: This study investigated the perceptions of alcohol and other drug (AOD) clinicians and consumers regarding the concepts of collaboration and empowerment. BACKGROUND/RATIONALE: Working collaboratively is seen as a cornerstone of empowerment, a concept used extensively over the last decade in an attempt to isolate power and control issues. It is generally assumed that health professionals strive to achieve collaboration, but little research has been undertaken. DESIGN/METHODS: The study was carried out in a variety of AOD treatment facilities in a large regional city in Australia. Facilities included an inpatient detoxification unit, an outpatients' clinic, a methadone maintenance clinic, a residential detoxification and rehabilitation unit using a 12-step treatment model and a residential unit using cognitive behavioural therapy (CBT). The nonprobability sample consisted of 57 staff and consumers who were interviewed using a semi-structured format. Data were analysed thematically using a Foucauldian analysis of discourse. RESEARCH FINDINGS: Findings suggest that clinicians may be unwittingly imposing their beliefs and prejudices on consumers and in fact disempowering the very people they want to empower. Collaboration is not happening. Foucault's theory of objectification illuminates the processes involved. CONCLUSIONS: There is much more happening than originally thought between consumers and staff when Foucault's three modes of objectification are used to examine this practice area. Analysing discourse enables us to see how health workers interact with consumers and the effect this may have on treatment objectives, outcomes and self-esteem. It also illuminates the work environment where the therapeutic encounter takes place. The need for further effort in empowering both consumers and staff and working collaboratively becomes evident. PMID- 11422544 TI - An examination of the role and function of psychiatric nurses in clinical practice in Ireland. AB - AIM OF THE STUDY: The aim of the study was to describe the role and function of all grades of psychiatric nurse in clinical practice so as to clarify the nature and scope of psychiatric nursing services. BACKGROUND AND RATIONALE: The psychiatric nursing role in Ireland and the United Kingdom (UK) in recent years has undergone a period of great change. There is a new emphasis on health promotion, early intervention, community development, with nursing being provided closer to where people live and work as well as making access to services easier for vulnerable groups of the population. Role ambiguity and the difficulty with defining psychiatric nursing work is a constant theme in the psychiatric nursing literature and this leaves the profession of psychiatric nursing rather vulnerable during this period of intense change. DESIGN AND METHODS: A descriptive qualitative research design with a multimethods approach to data collection was utilized involving three disparate but complimentary methods. RESULTS AND FINDINGS: Following data analysis, nine categories of nursing role were identified; these included both independent and interdependent roles. Clearly psychiatric nursing occupied a pivotal role in all mental health care settings. A major proportion of psychiatric nursing related to caring interactions and this appears to be a central nursing element. The assessment and maintenance of patient's safety was also important as mental health problems may place the patient or others in a position where their physical safety is threatened. CONCLUSION: It is concluded that Irish psychiatric nurses have been innovative and initiated many new services in response to emerging needs. Our understanding of psychiatric nursing is far from complete and the immediate challenge is to determine the knowledge and skills base required for independent therapeutic roles in response to changing mental health care needs. PMID- 11422545 TI - Using psychological insights to help people quit smoking. AB - AIMS: This paper sets out to evaluate the possibility that smoking cessation interventions which make use of current psychological theories and constructs can be more successful than programmes based largely on nicotine replacement therapy and will be more satisfying to participants. RATIONALE: Nicotine replacement therapy is currently the most widely used method for helping smokers give up the habit. Numerous studies have shown this to be a successful approach for many smokers, but the majority still fail to benefit. Typically three quarters of smokers given nicotine replacement are smoking again one year later. This study investigates whether nicotine replacement can be enhanced by the addition of psychological techniques. DESIGN: Smokers recruited via publicity in the local media were randomly assigned to one of two treatment conditions. The first condition consisted of a series of group sessions in which volunteers were instructed in nicotine replacement, and a number of psychological techniques, the most important being cognitive counter conditioning. The second condition was identical to the first but without the cognitive counter conditioning. Finally background quit rate was determined using waiting list controls. RESULTS: Both interventions were successful in helping smokers quit the habit, based upon an analysis at 6 months, compared with waiting list controls. The experimental condition incorporating cognitive counter conditioning produced a much higher quit rate than the condition based largely upon nicotine replacement, although the difference was not significant. CONCLUSIONS: This study is highly suggestive that nicotine replacement therapy can be enhanced by the inclusion of psychological techniques in group work, resulting in abstention rates higher than nicotine replacement alone and increasing participant satisfaction. Further work is needed with larger numbers to verify that this is indeed a significant gain and to investigate whether psychological techniques can give longer term benefits. PMID- 11422546 TI - Evaluating the self-assessed support needs of women with breast cancer. AB - AIMS OF THE STUDY: The first aim of the study was to identify the self-assessed support needs of women with breast cancer at various points of illness and, secondly, to establish if these needs formed clusters which could provide the basis for developing a standardized scale of needs for use by breast care teams in the evaluation of care. BACKGROUND: It has been found that support given to women with breast cancer has a positive effect upon their reactions to the illness and may even prolong their survival. Given that breast cancer affects a large number of women it is obviously important that those affected receive, in addition to the best available medical treatment, the type of support that best meets their needs. This study aimed to provide information on the impact of breast cancer and the need for various types of support by examining women's own assessment of their needs at different stages of their illness. DESIGN AND METHODS: A purposive sample of 12 women with a diagnosis of breast cancer was selected in one health authority in England, United Kingdom (UK). Women selected represented a wide age range (between 26 and 58), were married or in long-term cohabiting relationships and were at different points on the illness trajectory. Women were invited to take part in a semi-structured interview about their experiences of breast cancer. The data from these interviews were analysed using the software package Qualitative Solutions and Research, Nonnumerical Data Indexing, Searching and Theorizing (QSR*NUDIST). Following this content analysis, a questionnaire was formulated which divided statements into seven categories: diagnosis, treatment, support, femininity and body image, family and friends, information and after care, to be rated on a Likert scale ranging from "of no importance" to "extremely important". Questionnaire data were analysed by means of a one-way analysis of variance (for three independent variables) or t-test for two independent variables. Results. The questionnaire was sent to 971 women and achieved a response rate of 50.4%. The mean score for statements of need reached the level of point 4 on the Likert scale (important) with three exceptions: having professional help with family problems and domestic upheaval, coping with feelings of anger and dealing with the question "why me?" CONCLUSION: With the above three exceptions, women experienced a high level of need associated with a diagnosis of breast cancer. Studies of this kind should enable resources to be targeted to areas of highest need. PMID- 11422547 TI - Communication skills of nurses during interactions with simulated cancer patients. AB - AIM: In this paper the balance of affective and instrumental communication employed by nurses during the admission interview with recently diagnosed cancer patients was investigated. RATIONALE: The balance of affective and instrumental communication employed by nurses appears to be important, especially during the admission interview with cancer patients. METHODS: For this purpose, admission interviews between 53 ward nurses and simulated cancer patients were videotaped and analysed using the Roter Interaction Analysis system, in which a distinction is made between instrumental and affective communication. RESULTS: The results reveal that more than 60% of nurses' utterances were of an instrumental nature. Affective communication occurred, but was more related to global affect ratings like giving agreements and paraphrases than to discussing and exploring actively patients feelings by showing empathy, showing concern and optimism. CONCLUSION: In future, nurses should be systematically provided with (continuing) training programmes, in which they learn how to communicate effectively in relation to patients' emotions and feelings, and how to integrate emotional care with practical and medical tasks. PMID- 11422548 TI - Adherence to self-care and glycaemic control among people with insulin-dependent diabetes mellitus. AB - AIM OF THE STUDY: Factors associated with adherence to self-care and glycaemic control were studied in 213 people with insulin-dependent diabetes mellitus using a self-report questionnaire and a biochemical indicator (glycosylated haemoglobin). METHODS: The data were collected in the Oulu Health Center and the Central Hospital of Lapland in Northern Finland. The response rate was 76%. In order to verify the reliability and validity of the instruments, we used correlation coefficients, factor analysis and item-total analysis. Internal consistency was checked by Cronbach's alpha. The connections between self-care and the background variables were examined by cross-tabulation. FINDINGS: The majority of subjects accomplished their insulin treatment as scheduled, but had more difficulties with the other aspects of self-care. According to the findings, a fifth (19%) of the respondents were neglecting their self-care. The others undertook flexible (46%), regimen-adherent (16%) or self-planned self-care (19%). The subjects who were adherent to self-care had better metabolic control than those who neglected self-care. According to logistic regression analysis, poor metabolic control (P=0.003), smoking (P=0.009) and living alone (P=0.014) were associated with neglect of self-care. Gender, concurrent diseases and complications as a result of diabetes increased the risk, but had no significant association with adherence to or neglect of self-care. CONCLUSION: The findings demonstrated that adherence to self-care does not always lead to good metabolic control, but neglect of self-care is likely to lead to poor metabolic control. PMID- 11422549 TI - Health-related quality of life after myocardial infarction: an interview study. AB - AIM: The aim of this qualitative study was to explore and gain insights into the effects of myocardial infarction on health-related quality of life. BACKGROUND: The study was undertaken as part of a larger on-going project to develop and validate a disease-specific health-related quality of life instrument suitable for use with patients after myocardial infarction. METHODS: A consecutive sample of 31 patients admitted to a district general hospital in the North of England was recruited and interviewed at home. Semi-structured interviews were conducted based on a guide developed from a review of the literature pertaining to quality of life and expert opinion. Interviews were audio-tape recorded and transcribed verbatim. Transcripts were subjected to latent and manifest content analysis and inter-rater reliability was confirmed by a researcher not involved with the interview process. FINDINGS: Analysis of the data revealed seven major categories: physical activity/symptoms; insecurity; emotional reactions; dependency; lifestyle modification; concern over medication; and side-effects. Breathlessness, insecurity and feelings of over-protection were major problems, as was dissatisfaction with information and support. CONCLUSION: Myocardial infarction resulted in a variety of health-related problems which affected quality of life. Systematic monitoring and evaluation of health status should be performed routinely. This is likely to be aided by the development and use of a health-related quality of life instrument for this patient group. PMID- 11422550 TI - What do nurses and therapists think about the positioning of stroke patients? AB - BACKGROUND: At present, there are a number of different positioning strategies for stroke patients, but these are mainly based on clinical experience rather than research. Prior to developing a study to evaluate the effect of positioning on outcome after stroke, it was important to establish if nurses in our hospital had given much thought to the positioning of stroke patients. AIM OF THE STUDY: This study aimed to explore whether nurses working on the stroke unit at a Scottish teaching hospital held different views on the positioning of conscious and unconscious stroke patients to nurses working on other wards with stroke patients and therapists. METHODS: Questionnaires on various aspects of patient positioning were sent to 150 nurses and 25 therapists working in five specialities where stroke patients are cared for in a large teaching hospital. FINDINGS: Overall, the majority of nurses and therapists (74%) believed that the best position for conscious stroke patients was sitting in a chair. Also, 80% of them believed that the best position for unconscious stroke patients was lying on the nonparetic side. There was less of a consensus between nurses and therapists working in the five specialities as to whether it was appropriate for conscious or unconscious stroke patients to lie on their paretic side, lie supine or sit propped-up in bed in either a 30 or 70 degrees angle. CONCLUSIONS: The lack of consensus between nurses working in the five specialities is probably because at present there is little research to guide nursing practices for the positioning of stroke patients. Therefore, research to confirm which positions improves or hinders outcome after stroke is essential. Indeed, positioning is a simple inexpensive strategy, which could have a substantial public health impact, as stroke is so common. PMID- 11422551 TI - Patterns of elderly spousal caregiving in dementia care: an observational study. AB - AIM: The aim of this study was to describe which caring activities eight spouses performed when caring for a partner with dementia, and in what way these activities were carried out. BACKGROUND: Family caregivers are recognized as being the primary source of care for the community's older people. The largest group is comprised of spouses, with wives as the predominant caregivers. This informal care seems to be more or less invisible and performed in silence within the family. Despite the wealth of studies, the essence of family caregiving is not well understood. METHODS: Data collection was conducted by observing the dyads in their homes. A qualitative approach inspired by grounded theory was chosen to discover qualities and describe patterns of spousal caregiving in dementia care. RESULTS: The analysis yielded four broad themes, which included nine categories. Findings from the study shed some light on the invisible aspects besides the traditional hands-on caregiving. CONCLUSION: The elderly carers were engaged in demanding and time-consuming care ranging from supervision to heavy physical responsibility. They were caring for as well as about their partners. The study also showed that spouses were successful in managing their situation in different ways. The results reported in this article are unique as they come from direct observations in family home settings where a spouse cared for a partner with dementia. Knowledge about family caregiving is valuable for nurses as there is an emphasis on collaboration between family caregivers and professionals. PMID- 11422552 TI - Nurse-led intermediate care: an opportunity to develop enhanced roles for nurses? AB - BACKGROUND: Nurse-led intermediate care units are being set up across the UK primarily as potential solutions to hospital bed crises. AIMS: This paper draws on data collected as part of a comprehensive evaluation of one 10-bedded nurse led unit (NLU) located in the South of England. It explores the potential for enhanced nursing roles provided by such units by focusing on the views of NLU nursing staff and other professional groups within the Hospital Trust where the unit is located. METHODS: A total of 38 in-depth audio-taped qualitative interviews were conducted with NLU nursing staff and with a range of other professional groups (managers, acute ward nurses and doctors). FINDINGS: These data indicated that models of nurse-led postacute care do provide opportunities for nurses to develop enhanced nursing roles in which care associated with concepts of therapeutic nursing can be provided. However, even though the nurses derived satisfaction from their work on the NLU this model of care was seen by junior and middle grade nurses and other professional groups as being of low status. In contrast to senior nurses' views, they did not equate work on the NLU with the continuing professionalization of nursing. Senior nurses viewed the route to developing nursing on the NLU as involving nurses as doctor substitutes (extended roles) rather than as working in separate but complementary therapeutic domains (enhanced roles). CONCLUSIONS: NLUs provide opportunities for nurses to develop enhanced roles in which they can work autonomously in providing elements of therapeutic nursing aimed at improving patient outcomes at discharge. However, education, training and leadership will be needed to ensure that such opportunities are well understood and are optimized to the benefit of nurses and their patients. PMID- 11422553 TI - Health education, behavioural change and social psychology: nursing's contribution to health promotion? AB - AIMS: To critically review the complex processes that underpin the modification of a client's health-related behaviour. This paper also seeks to contextualize the operational differences between health-educating and health-promoting activities - as a means of rationalizing current practice. BACKGROUND: In "health promotional" encounters, there is a plethora of evidence that suggests that nurses work predominantly within a "traditional" preventative framework of practice. The prevalence of a culturally inherent biomedical framework, governing most nursing practice, tends to reduce health-related client interventions to little more than one-off, reductionist information-giving exercises. The expectation on clients to respond to and subsequently modify their health behaviour, when presented with such information, is unrealistic in most cases. Nurses are often unaware of the extremely complex human phenomena associated with modifying health-related behaviours and the resultant change processes. In nursing-related health encounters, the planned or unplanned intervention and the subsequent outcomes are mostly viewed within a too simplistic and superficial context. DESIGN: A selective review of the relevant literature. CONCLUSION: Where many nurses believe themselves to be health promotionalists, the likelihood is that they are instead more likely to be traditional health educationalists. Not that this is the main problem, in itself--but if nursing is to progress on this issue, it must first become more effective in delivering its current health education initiatives. Armed with further knowledge and understanding of their practices, health educators are far more likely to achieve a degree of success in their behavioural-change encounters as well as approach the intervention with a far more realistic expectation of outcome. Without this further understanding, it is argued that the integration of health educational initiatives into nursing practice will generally do little or nothing to change the health status of clients. PMID- 11422554 TI - Spiritual coping strategies: a review of the nursing research literature. AB - AIMS OF THE PAPER: This paper reviews some of the limited nursing research-based literature, orientated towards the use of spiritual coping strategies in illness. This review aims at identifying those spiritual coping strategies used by the believers and nonbelievers followed by implications for holistic nursing care. LITERATURE SEARCH: The CINAHL and MEDLINE CD Rom databases were searched, identifying literature published from 1975 onwards which amounted to 187 articles. The majority of the literature traced were found anecdotal with only few studies investigating directly spiritual coping strategies. Following scrutiny of the available articles, only five research studies explored directly the spiritual coping strategies used in various illness, four of which were conducted in USA and one in UK. Because of the small scale research studies, generalization of the findings of this review is limited to the samples used. THEORETICAL BACKGROUND: Research suggests that spiritual coping strategies, involving relationship with self, others, Ultimate other/God or nature were found to help individuals to cope with their ailments. This may be because of finding meaning, purpose and hope, which may nurture individuals in their suffering. Spirituality is oftenly referred by literature as being synonymous with religiosity. Thus the use of spiritual coping strategies is restricted to individuals who hold religious beliefs. However, the definition of spirituality indicates that this concept is broader than religiosity. The theories on stress coping (Folkman & Lazarus 1984) and the numinous experience (Otto 1950) outline the rationale for the use of these strategies which are applicable to both the believers and nonbelievers. IMPLICATIONS: This review suggests that the onset of illness may render the individual, being a believer or nonbeliever to realize the lack of control over his/her life. However the use of spiritual coping strategies may enhance self-empowerment, leading to finding meaning and purpose in illness. This implies that holistic care incorporates facilitation of various spiritual coping strategies to safeguard the wholeness and integrity of the patients. PMID- 11422555 TI - Changes, challenges and choices for the primary health care workforce: looking to the future. AB - AIMS: The main aim of this paper is to draw attention to problems facing the primary health care workforce in terms of demand for treatment of minor illness over the next two decades. These predictions have implications for the community nursing workforce in particular and the flexibility of primary health care teams in general. BACKGROUND: Care delivered in the primary care sector influences, and is influenced by, the characteristics of the health care workforce. These characteristics fall into two main groups: firstly, the shape of the present medical and nursing workforce and manpower trends; and secondly, the changes in doctors' and nurses' workloads. DESIGN: This paper draws on two studies, both commissioned by the Department of Health; the first study focusing on skill mix and delegation in primary health care teams and the second addressing the implications of skill mix for medical workforce scenarios in the changing policy environment. FINDINGS: From the first study, general practitioners across ten general practices were prepared to delegate at least one topic from over a third of 836 consultations and a further 17% of entire consultations. This potential delegation fell mainly to practice nurses and nurse practitioners. The second study used data extracted from the National Morbidity Surveys of 1981 and 1991 predicting that minor consultations are set to increase by 11 million from the 1990s to 2020--a minimal estimate. The authors argue that many of these extra predicted consultations will find their way onto practice nurses' and nurse practitioners' caseloads. CONCLUSIONS: Workforce issues and questions of professional roles and boundaries, in the context of the "greying" community nursing workforce, demand solutions if patient/client demand is to be met over the next two decades. PMID- 11422556 TI - Taxonomy of Campylobacter, Arcobacter, Helicobacter and related bacteria: current status, future prospects and immediate concerns. PMID- 11422557 TI - The physiology and metabolism of Campylobacter jejuni and Helicobacter pylori. PMID- 11422558 TI - Surface components of Campylobacter and Helicobacter. PMID- 11422559 TI - Post genome analysis of Campylobacter jejuni. PMID- 11422560 TI - Pathogenesis of enteric Campylobacter infection. PMID- 11422561 TI - Animal models of Campylobacter jejuni colonization and disease and the lessons to be learned from similar Helicobacter pylori models. PMID- 11422562 TI - Campylobacters in water, sewage and the environment. PMID- 11422563 TI - Helicobacter in water and waterborne routes of transmission. PMID- 11422564 TI - Current epidemiological issues in human campylobacteriosis. PMID- 11422565 TI - Poultry as a source of Campylobacter and related organisms. PMID- 11422566 TI - Campylobacter spp. in the kitchen: spread and persistence. PMID- 11422567 TI - Practical intervention strategies for Campylobacter. PMID- 11422568 TI - Helicobacter pylori: a clinical update. PMID- 11422569 TI - Antibiotic resistance of Helicobacter pylori. PMID- 11422570 TI - Guillain--Barre syndrome and Campylobacter jejuni infection. PMID- 11422572 TI - Methyl orange antagonizes uridine 5' triphosphate and not alpha,beta-methylene adenosine 5' triphosphate-evoked depolarization of rat superior cervical ganglia. AB - 1. Compared with the effects of adenosine 5' triphosphate (ATP) on the nervous system, the actions of pyrimidine nucleosides and their 5'-nucleotides, such as uridine 5' triphosphate (UTP), have received less attention. In part, this is because there is a need for a selective antagonist for responses mediated by UTP activated receptors. The objective of this study was to discover such an antagonist. 2. Superior cervical ganglia isolated from male rats were superfused with a physiological salt solution. Responses to alpha,beta-methylene-ATP (alpha,beta-Me-ATP), potassium, adenosine and UTP were determined before and in the presence of 1-300 microM methyl orange. 3. Methyl orange at 1-100 microM did not alter resting potential or depolarizing responses to alpha,beta-Me-ATP, potassium, or adenosine-evoked hyperpolarizations, but at 10 and 100 microM methyl orange significantly antagonized UTP-evoked depolarizations (P < 0.05). 4. Although the antagonistic effects of methyl orange were not dramatic, this is the first report of a putative pyrimidinoceptor antagonist. These observations also support the idea of distinct receptors for UTP and ATP on rat superior cervical ganglia. PMID- 11422573 TI - Vascular responses to beta-adrenoceptor subtype-selective agonists with and without endothelium in rat common carotid arteries. AB - 1. Using the cannula inserting method, vasodilator responses to beta-adrenoceptor agonists (isoprenaline, denopamine and procaterol) were investigated in isolated and perfused rat common carotid arteries. 2. Each beta-adrenoceptor agonist induced a vasodilation in preparations preconstricted by phenylephrine in a dose related manner. The potencies were in the order of isoprenaline > procaterol >> denopamine. 3. Denopamine-induced dilations were significantly inhibited by 1 nmol betaxolol (a selective beta1-adrenoceptor antagonist), but it was not influenced by 1 nmol ICI 118,551 (a selective beta2-adrenoceptor antagonist). On the other hand, procaterol-induced vasodilations were significantly inhibited by 1 nmol ICI 118,551 but not modified by 10 nmol betaxolol. 4. ACh-induced vasodilations disappeared after intraluminal saponin injection to remove endothelium, but procaterol- and denopamine-induced dilations were not modified by removal of the endothelium. 5. Pretreatment with L-NG-nitroarginine methyl ester (L-NAME) readily inhibited ACh-induced vasodilations. However, neither procaterol- or denopamine-induced vasodilation was modified by L-NAME treatment. 6. From these results, it is concluded that in the rat common carotid arteries (1) there are abundant beta2- and a few beta1-adrenoceptors, and (2) there is no participation of the endothelium-dependent mechanism in beta-adrenoceptor mediated vasodilations. PMID- 11422574 TI - The effect of streptozotocin-induced diabetes on cardiac beta-adrenoceptor subtypes in the rat. AB - 1. The present study investigates the effect of short-term experimental diabetes of 14-days duration on the beta-adrenoceptor subtypes of the rat heart. 2 .beta adrenoceptor-mediated functional responses to submaximal doses of isoprenaline were enhanced in Langendorff-perfused hearts from diabetic rats, manifested as greater changes in tension, heart rate and rates of tension development (+dT/dt) and decline (-dT/dt). 3. Radioligand binding data demonstrated that total cardiac beta-adrenoceptor density and affinity for [3H]-dihydroalprenolol was unchanged by diabetes, although a decrease in beta1-adrenoceptor density and increase in beta2-adrenoceptor density was observed. 4. In conclusion, hearts from 14-day streptozotocin-induced diabetic rats demonstrate a number of alterations within the beta-adrenoceptor system. However, the enhanced beta-adrenoceptor-mediated responses to isoprenaline were not caused by an overall increase in density of beta-adrenoceptors, but were accompanied by changes in the ratio of the beta adrenoceptor subtypes. PMID- 11422575 TI - The effects of thiol compounds and ebselen on nitric oxide activity in rat aortic vascular responses. AB - 1. Thiols have been implicated to play a role in a variety of aspects of nitric oxide (NO) generation and activity. Thiol dependence of nitric oxide synthase (NOS) has remained controversial and its mechanism is not clear. This study investigates possible mechanisms between thiol (SH group) and NOS activation, through thiol compounds (glutathione, dithiothreitol, N-acetyl-L-cysteine) and Ebselen [2-phenyl-1,2-benzisoselenazole-3(2H)-one] on rat aortic vascular responses. 2. In rat thoracic aorta, acetylcholine (10(-6)-10(-9) M) induced a relaxation of phenylephrine (PE) (10(-7) M)-induced tone, which was inhibited dose dependently by increasing concentration of ebselen (1-10 microM). 3. In rings of rat thoracic aorta, ebselen and NOS inhibitors (NG-monomethyl-L arginine, NG-nitro-L-arginine methyl ester) produced an augmentation of phenylephrine (10(-7) M)- induced tone and acetylcholine induced a relaxation of PE (10(-7) M)-induced tone in rat thoracic aorta, which was inhibited by ebselen (10 microM) like NOS inhibitor. 4. The thiol compounds (glutathione, dithiothreitol, and N-acetyl-L-cysteine) alone did not change vascular tone in rat thoracic aorta. Pretreatment with thiol compounds before ebselen treatment, however, reversed the inhibitory effect of ebselen which acts like the NOS inhibitor in rat thoracic aorta. Posttreatment with thiol compounds after ebselen treatment did not reverse the inhibitory effect of ebselen by as much as pretreatment. 5. Calcium ionophore A23187 (10(-7) M)-induced vasodilation was inhibited in ebselen pretreated rat thoracic aorta, but sodium nitroprusside (SNP, 10(-7) M)-induced relaxation was not inhibited by ebselen. This suggests that NOS is involved in the inhibitory effect of ebselen on rat thoracic aorta relaxation. 6. These results suggest that ebselen exerts an inhibitory action on the nitric oxide synthesis in rat thoracic aorta by interacting with thiol groups. PMID- 11422576 TI - Activation of presynaptic A1-receptors by endogenous adenosine inhibits acetylcholine release in the guinea-pig ileum. AB - 1. It is well established that presynaptic adenosine A1-receptor activation inhibits acetylcholine (ACh) release in the guinea-pig ileum. The present study extends this observation and examines a possible role for endogenous adenosine in modulating cholinergic nerve function. 2. The actions of the adenosine uptake blocker, dipyridamole, the adenosine deaminase inhibitor, erythro-9(2-hydroxy-3 nonyl)adenine (EHNA) and the A1-receptor antagonist, 1,3-dipropyl-8 cyclopentylxanthine (DPCPX) were examined on electrically evoked neurogenic, cholinergic twitch contractions of the guinea-pig ileum. Some additional studies measuring [3H]-ACh release were also performed. 3. Adenosine and the selective A1 receptor agonist, 2-chloroadenosine (2-CA), inhibited electrically evoked contractions and, in the case of 2-CA, [3H]-ACh release. The actions were antagonized by DPCPX. At low concentrations, dipyridamole and EHNA enhanced the effect of adenosine causing a leftward shift of the concentration-response curve. In contrast, inhibition induced by 2-CA was unaffected by either dipyridamole or EHNA. 4. When applied alone at higher concentrations, EHNA and dipyridamole produced a concentration-dependent suppression of cholinergic neurotransmission. In both cases, the effect could be reversed by DPCPX. At the same concentration, DPCPX alone produced a small but consistent increase in twitch height and [3H] ACh release. 5. The data confirm the existence of inhibitory presynaptic adenosine A1-receptors modulating cholinergic nerve function in the guinea-pig ileum and suggests that these receptors can be activated by endogenous adenosine released either as adenosine itself or as an ATP metabolite. PMID- 11422577 TI - Arrhythmogenesis in isolated rat hearts with enhanced alpha-adrenoceptor-mediated responsiveness. AB - 1. It has been postulated that stimulation of myocardial alpha-adrenoceptors is one of the primary mediators of the dysrhythmias which occur during periods of myocardial ischaemia and reperfusion. This study examines arrhythmogenesis during coronary artery occlusion and reperfusion in isolated perfused rat hearts from control animals and from rats with enhanced myocardial alpha-adrenoceptor responsiveness. 2. Rats were administered propylthiouracil (PTU) in their drinking water for 8 weeks. This treatment resulted in an enhanced responsiveness of isolated left atria to the alpha-adrenoceptor agonist phenylephrine compared with atria from control animals. 3. In Langendorff-perfused isolated hearts, the spontaneous rate of contraction was significantly lower in the PTU-pretreated group than in either age-matched or weight-matched controls. Occlusion of the left anterior descending artery (LAD) for 25 min resulted in ventricular tachycardia (VT) of similar incidence and duration in all groups and ventricular fibrillation (VF) in both control groups but not the PTU-pretreated group. 4. Following the 25-min ischaemic period the myocardium was reperfused for 10 min. The incidence and duration of VT and VF during this period was similar in all groups except that the duration of VF in the PTU-pretreated group was significantly lower than in controls. 5. In perfused hearts paced at 4 Hz, the incidence and duration of dysrhythmias during ischaemia and reperfusion was again similar in all groups, only the duration of VF being affected (reduced) by PTU pretreatment. 6. In conclusion, this study does not lend support to the hypothesis that myocardial alpha-adrenoceptors have a primary role in arrhythmogenesis, but the data would support a role for these receptors in myocardial protection. PMID- 11422578 TI - Presynaptic alpha2-adrenoceptor-mediated modulation of adenosine 5' triphosphate and noradrenaline corelease: differences in canine mesenteric artery and vein. AB - 1. The modulatory effects of agonists and antagonists of prejunctional alpha2 adrenoceptors on the electrical field stimulation (EFS, 0.3 ms, 12 V)-induced release of endogenous noradrenaline (NA) and the cotransmitter adenosine 5' triphosphate (ATP) were measured in endothelium-denuded segments of canine inferior mesenteric artery and compared with effects in mesenteric vein. The overflow of NA and ATP was evoked by long-duration (2 min) EFS at low frequency (4 Hz) and high frequency (16 Hz) of stimulation and was analysed using HPLC techniques with electrochemical detection and fluorescence detection, respectively. 2. The EFS-evoked overflow of both NA and ATP was significantly reduced by tetrodotoxin (1 microM) and guanethidine (10 microM) in the artery and vein. Desipramine (10 microM), a blocker of neuronal uptake of NA, increased the EFS (4 and 16 Hz)-evoked overflow of NA in both artery and vein. EFS-evoked overflow of NA in vein exceeded the NA overflow in artery at both 4 and 16 Hz in control preparations as well as in the presence of desipramine. However, the EFS evoked overflow of ATP was equal in the artery and vein. 3. Stimulation of alpha2 adrenoceptors with clonidine (0.1 microM) and oxymethazoline (0.3 microM) reduced the EFS evoked overflow of NA in both artery and vein at 4 Hz, whereas the NA overflow at 16 Hz remained unchanged in both blood vessels. The overflow of ATP as well as of ADP (and hence ATP:ADP ratio) was unaffected by the alpha2 adrenoceptor agonists in the artery and vein. 4. In artery, blockade of alpha2 adrenoceptors with yohimbine at a concentration of 0.1 microM caused no effect on the NA overflow neither at 4 Hz nor at 16 Hz of EFS. Yohimbine at a concentration of 1 microM increased the overflow of NA at 4 Hz but not 16 Hz of EFS. In vein, however, yohimbine (0.1 and 1 microM) increased NA overflow at both 4 and 16 Hz of stimulation. Idazoxan (1 microM) increased the NA overflow in artery only at 4 Hz, whereas in vein idazoxan increased the NA overflow at both 4 and 16 Hz. No changes of EFS-evoked ATP overflow were observed in the presence of 0.1 microM yohimbine in both artery and vein. Greater concentration of yohimbine (i.e. 1 microM) increased the overflow of ATP in both the artery and vein only at 4 Hz EFS. Idazoxan (1 microM) enhanced the ATP overflow only at 16 Hz in vein. The overflow of ADP was affected by both yohimbine and idazoxan in a similar manner to the ATP overflow so that the ATP:ADP ratios were not changed. 5. In conclusion, sympathetic nerves in both mesenteric arteries and veins appear to release ATP along with NA. Release of NA in veins exceeds release of NA in arteries, whereas both the canine artery and vein release equal amount of ATP. At long-duration nerve stimulation (as might occur during stress) the alpha2 adrenoceptors appear to rather modulate release of NA than release of the cotransmitter ATP. The prejunctional autoinhibition of NA release is more effective at lower frequencies of nerve stimulation. The alpha2-adrenoceptor mediated neuromodulation plays a greater role in veins than arteries. Quantitative differences in alpha2-adrenoceptor-mediated neuromodulation in the arteries and veins may participate to differing contributions of mesenteric blood vessels to the control of blood flow and volume distribution in splanchnic circulation. PMID- 11422579 TI - Modulation of M-channel conductance by adenosine 5' triphosphate in bullfrog sympathetic B-neurones. AB - 1. Adenosine 5' triphosphate (ATP) (0.5-500 microM) or muscarine (0.1-1.0 microM) suppressed M-current/conductance (IM/gM) in B-cells of bullfrog sympathetic ganglion. Both agonists suppressed steady-state M-conductance (gM) at -30 mV and there was either no change or a slight increase in the time constants for gM activation (tau(a) at -30 mV) and deactivation (tau(d) at -50 mV). 2. It has previously been shown that experimental elevation of intracellular Ca2+ concentration ([Ca2+]i) suppresses gM and this is associated with decreases in both tau(a) and tau(d). As these changes in kinetics differ from those we observe with agonist application, our results cast doubt on the hypothesis that elevation of [Ca2+]i is involved in the transduction mechanism for ATP- or muscarine induced gM suppression. PMID- 11422580 TI - Change in subgingival microbial profiles in adult periodontitis subjects receiving either systemically-administered amoxicillin or metronidazole. AB - AIM: The current investigation evaluated changes in levels and proportions of 40 bacterial species in subgingival plaque samples during, immediately after and up to 1 year after metronidazole or amoxicillin therapy combined with SRP. METHOD: After baseline clinical and microbiological monitoring, 17 adult periodontitis subjects received full mouth SRP and 14 days systemic administration of either metronidazole (250 mg, TID, n=8) or amoxicillin (500 mg, TID, n=9). Clinical measurements including % of sites with plaque, gingival redness, bleeding on probing and suppuration, pocket depth (PD) and attachment level (AL) were made at baseline, 90, 180 and 360 days. Subgingival plaque samples were taken from the mesial surface of all teeth in each subject at baseline, 90, 180 and 360 days and from 2 randomly selected posterior teeth at 3, 7, and 14 days during and after antibiotic administration. Counts of 40 subgingival species were determined using checkerboard DNA-DNA hybridization. Significance of differences over time was determined using the Quade test and between groups using ANCOVA. RESULTS: Mean PD was reduced from 3.22+/-0.12 at baseline to 2.81+/-0.16 (p<0.01) at 360 days and from 3.38+/-0.23 mm to 2.80+/-0.14 mm (p<0.01) in the amoxicillin and metronidazole treated subjects respectively. Corresponding values for mean AL were 3.21+/-0.30 to 2.76+/-0.32 (p<0.05) and 3.23+/-0.28 mm to 2.94+/-0.23 mm (p<0.01). Levels and proportions of Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola were markedly reduced during antibiotic administration and were lower than baseline levels at 360 days. Counts (x10(5), +/-SEM) of B. forsythus fell from baseline levels of 0.66+/-0.16 to 0.04+/-0.02, 0.13+/-0.04, 0.10+/-0.03 and 0.42+/-0.19 in the amoxicillin group at 14, 90, 180 and 360 days respectively (p<0.001). Corresponding values for metronidazole treated subjects were: 1.69+/-0.28 to 0.02+/-0.01, 0.20+/-0.08, 0.22+/-0.06 and 0.22+/-0.08 (p<0.001). Counts of Campylobacter species, Eubacterium nodatum, Fusobacterium nucleatum subspecies, F. periodonticum and Prevotella nigrescens were also detected at lower mean levels during and immediately after therapy, but gradually increased after withdrawal of the antibiotics. Members of the genera Actinomyces, Streptococcus and Capnocytophaga were minimally affected by metronidazole. However, amoxicillin decreased the counts and proportions of Actinomyces species during and after therapy. CONCLUSIONS: The data suggest that metronidazole and amoxicillin are useful in rapidly lowering counts of putative periodontal pathogens, but must be accompanied by other procedures to bring about periodontal stability. PMID- 11422581 TI - In vitro effect of oral antiseptics on human immunodeficiency virus-1 and herpes simplex virus type 1. AB - AIM: The antiviral effectiveness of widely used commercial mouthrinses has not been well studied. A project was undertaken to evaluate and compare the in vitro antiviral effectiveness of essential oil-containing mouthrinses (LA & TLA) and chlorhexidine mouthrinses (PX & CHX) on 2 different enveloped viruses, human immunodeficiency virus (HIV-1) and Herpes simplex virus (HSV-1) McIntyre strain. METHOD: HIV-1(89.6) (1x10(5)/ml) and HSV-1 (1x10(6)/ml) in RPMI-1640 medium were treated with two commercially available forms of LA & TLA (tartar control LA), and 2 formulations of chlorhexidine [(PX), 0.12% chlorhexidine & (CHX), 0.2% chlorhexidine] for 30 sec. The antiviral effect was estimated by inhibition of the syncytia formation or the cytopathic effect (CPE) for HIV-1 on MT-2 cells and by inhibition of the plaque formation for HSV-1 on Vero cell monolayers. RESULTS: Undiluted LA, TLA, PX and CHX completely inhibited both HIV-189.6 and HSV-1 McIntyre strain. PX and CHX inhibited HIV-1 up to 1:4 dilution, whereas, LA and TLA inhibited HSV-1 up to 1:2 dilution. The antiviral effects of LA and TLA were found to be similar and also the antiviral effect of PX and CHX were also found to be comparable. CONCLUSIONS: The methods used in this investigation allow easy and reproducible evaluations of antiviral efficacy. The anti-HIV-1 and anti-HSV-1 effects of LA, TLA, PX and CHX as evidenced in our in vitro study suggest that we should investigate potential in vivo effects during the use of essential oil containing or chlorhexidine containing products when used by patients as mouthrinses. If the clinical studies confirm the in vitro data, pre-procedural use by clinicians may be beneficial in reducing viral contamination of bio aerosols during the delivery of dental care. PMID- 11422582 TI - Untreated periodontal disease in Indonesian adolescents. Subgingival microbiota in relation to experienced progression of periodontitis. AB - BACKGROUND/AIMS: In an Indonesian population deprived of regular dental care, the experienced progression of disease between baseline (1987) and follow-up (1994) was investigated in relation to the composition of the subgingival microbiota at follow-up. At baseline the age ranged from 15 to 25 years. Clinical and microbiological evaluation was completed in 158 of the 167 subjects available at follow-up. METHODS: Plaque index (PI), pocket depth (PD), bleeding on probing (BOP), and attachment loss (AL) were scored at the approximal surfaces of all teeth and subgingival calculus on the approximal surfaces of the Ramfjord teeth only (number of sites with subgingival calculus: NSC). A pooled sample of the deepest pocket in each quadrant was evaluated using microbiological culture techniques. RESULTS: At baseline the mean values of the clinical parameters were AL=0.35 mm, PI=1.01, BOP=0.80 PD=3.25 mm and NSC=6.04 and at follow-up AL=0.75 mm, PI=1.16, BOP=1.19, PD=3.34 mm and NSC=5.85. All parameters except PD and NSC showed a statistically significant increase. At follow-up the prevalence of Actinobacillus actinomycetemcomitans was 40%, of Porphyromonas gingivalis 67%, of Prevotella intermedia 66%, of Fusobacterium nucleatum 79%, of Bacteroides forsythus 16%, of Campylobacter rectus 4%, and of P. micros 6%. No differences in clinical parameters were found between groups with or without these micro organisms. In 129 subjects AL of > or =2 mm at > or =1 site was found. Logistic regression showed three significant odds-ratio's for experienced progressive periodontitis: Plaque index (12.2), gender (3.4) and Actinobacillus actinomycetemcomitans (2.9). CONCLUSIONS: The results of this retrospective study suggest that plaque is the most important parameter related to experienced disease progression, and that the presence of A. actinomycetemcomitans may be associated with increased chance of disease progression. PMID- 11422583 TI - Marginal bone loss around implants retaining hinging mandibular overdentures, at 4-, 8- and 12-years follow-up. AB - AIM: The aim of the present study was to study the effect of occlusal factors, oral hygiene, gender and age on the long-term maintenance of alveolar bone height around screw-shaped machined surface commercially pure (c.p.) titanium implants retaining hinging mandibular overdentures. METHOD: In this retrospective study a long-term (4-, 8-, up to 12-years) follow-up of such implant-retained overdentures was performed. From the population of patients so treated at the University Hospital of the Catholic University Leuven, Department of Periodontology and Department of Prosthetic Dentistry, 158 patients were selected on the basis of being treated before March 1994, and the availability of intra oral radiographs (paralleling technique) present at abutment connection (baseline) and at years 4, 8, and 12. For each patient, all relevant clinical data were recorded, and measurements of marginal bone height around the implants were performed on intra-oral radiographs by means of a digital sliding caliper. RESULTS: Results indicated that only the factor "time" had a significant influence on marginal bone loss. The age and gender of the patients, dental/prosthetic status in the antagonistic jaw, oral hygiene variables, and location of the implants had no significant effect. CONCLUSION: The very high cumulative success rate (97.2%) and limited marginal bone loss (on average 1.7 mm) after 12-years, encourage this kind of treatment using this type of implants. PMID- 11422584 TI - A multifactorial investigation of the ability of oral health care products (OHCPs) to alleviate oral malodour. AB - AIM, BACKGROUND: Oral malodour (halitosis) is generally ascribable to oral microbial putrefaction generating malodorous volatile sulphur compounds which predominantly comprise dihydrogen sulphide and methyl mercaptan. This study assesses the relative effectiveness of 6 oral health care products in reducing oral cavity volatile sulphur compound concentrations. METHOD: A mixed model 3 factor factorial experimental design involving 6 volunteers, 7 treatment regimens (products I-VI* and water placebo) and 5 time-points (0.00-5.29 h) was undertaken. Electron-donating volatile sulphur compound levels were determined in triplicate using a sulphide monitor (Interscan model 1170) both prior to (0.00 h) and following oral rinsing (20 ml of 5 of the products) or chewing (2 capsules of the remaining product) episodes with each product examined (0.29, 1.29, 2.29 and 5.29 h post-administration). RESULTS: Results were recorded as peak and steady state volatile sulphur compound equivalents (ppb). With the exception of one of the products, each oral health care product tested was found to reproducibly reduce volatile sulphur compound concentrations within 20 min of treatment; the mean % decreases in peak (and corresponding steady-state) levels ranging from 3.6 (0.0) to 16.8 (16.4)%. Subsequently, volatile sulphur compound concentrations returned to their zero-control (baseline) values within 5 h, the rate of this regression being in the reverse of the order observed for the magnitude of the primary 20 min reduction for both peak and steady-state measurements. As expected, the water placebo exerted no influence on oral cavity volatile sulphur compound levels. The most effective oral health care products contained admixtures of chlorite anion and chlorine dioxide (both of these agents have the ability to directly oxidise volatile sulphur compounds to non-malodorous products and the latter is also powerfully cidal towards odourigenic micro-organisms). CONCLUSIONS: We therefore conclude that oral health care products containing such oxohalogen oxidants may provide a useful therapeutic strategy for the treatment of oral malodour. PMID- 11422585 TI - A comparative in vitro study of a magnetostrictive and a piezoelectric ultrasonic scaling instrument. AB - BACKGROUND: The effects of magnetostrictive ultrasonic instruments and piezoelectric ultrasonic instruments on tooth surfaces seem to differ with regards to root debridement. AIM: The purpose of this study was to compare a magnetostrictive ultrasonic scaling instrument with a piezoelectric ultrasonic scaling instrument and a hand curette regarding time taken, calculus removal, tooth surface roughness (Ra), and SEM examination before and after instrumentation. METHODS: 30 extracted human teeth with subgingival calculus were assigned to one of three treatment groups (n=10). The working force was standardised for both ultrasonic instruments at 200 g and for the curette at 500 g. RESULTS: The results revealed that the time needed for instrumentation was 126.1+/-38.2 s for the curette, significantly more than for the piezoelectric ultrasonic instrument (74.1+/-27.6 s; p<0.05) and 104.9+/-25.4 s for the magnetostrictive ultrasonic instrument. Remaining calculus was similar for all three groups. The end Ra values were significantly worse for the piezoelectric instrument (2.02+/-0.41; p<0.05) compared to 1.42+/-0.48 for the curette and 1.36+/-0.41 for the magnetostrictive instrument. The SEM examination revealed the smoothest surfaces but, subjectively, the most tooth substance loss after the curette, followed by the magnetostrictive instrument, with the least substance loss, and then the piezoelectric instrument, with medium substance loss. CONCLUSION: The piezoelectric ultrasonic scaler was more efficient than the magnetostrictive ultrasonic scaler in removing calculus but left the instrumented tooth surface rougher. PMID- 11422586 TI - CD9 and HLA-DR expression by crevicular epithelial cells and polymorphonuclear neutrophils in periodontal disease. AB - BACKGROUND, AIMS: The composition of gingival crevicular fluid (GCF) is likely to reflect inflammatory modifications that take place in the gingiva during periodontal diseases. METHOD: In this study, GCF was collected at 3 different sites from 23 periodontal patients. The sites were assessed to be healthy, presenting gingivitis or periodontitis. 10 healthy individuals without any form of periodontal disease formed the control group and were sampled at one site each. The cell content of GCF was collected using Durapore Millipore strips, and 2 types of cells were studied: epithelial cells (EC) and polymorphonuclear neutrophils (PMN). The expression of CD9 and HLA-DR within or on the surface of these cells was studied in immunofluorescence on cytospin smears. RESULTS: Both CD9 and HLA-DR expression on EC differed significantly from control subjects, and the latter decreased according to the severity of the pathology. None of the PMN found in controls expressed CD9 or HLA-DR. However, in periodontal patients, the expression of HLA-DR within PMNs was detectable and increased according to the severity of lesions. CD9 expression on PMNs also increased with inflammation. CONCLUSION: This study shows that clinically healthy sites of periodontal patients already present signs of immunological activation characterised by a down modulation of HLA-DR expression on EC and an upregulation of these 2 molecules in PMN. PMID- 11422587 TI - Human histology of new attachment after root coverage using subepithelial connective tissue graft. AB - BACKGROUND, AIMS: Connective tissue grafts are used successfully in periodontal therapy for root coverage. However, reports on the histologic interface between the root surface and the grafted tissue have been few in number. This report describes a case study in which a subepithelial connective tissue graft was placed in a 27-year-old female on the maxillary left side. METHODS: The graft (15 mm long, 10 mm wide, 1.5 mm thick) included palatal periosteum and was placed with the periosteal side facing the exposed bone and root surfaces. RESULTS: 15 weeks after grafting, the teeth presented with residual recessions of 1 mm, and buccal probing depths were approximately 1 mm. 14 months post-surgery, the 1st maxillary premolars on both sides were extracted for orthodontic therapy. Clinical parameters at the graft site remained as at 15 weeks. Histologic analysis of tooth #24 showed that the sulcular epithelium was keratinized; epithelium lining the dentin exhibited rete ridges projecting into the gingival connective tissue; and junctional epithelium extended over new cementum. New connective tissue attachment was also observed, including periodontal ligament. CONCLUSION: Biological width was comparable pre- and post-surgery, indicating a real gain in attachment of 3.9 mm. PMID- 11422588 TI - Periodontal disease status of an indigenous population of Guatemala, Central America. AB - OBJECTIVES: This study was designed to determine the periodontal disease status of an indigenous Indian community of rural Central America (San Juan La Laguna, Guatemala), for comparison with results of similar studies in other populations, and with a view to performing future studies to address familial clustering of adult periodontitis. METHODS & RESULTS: An initial screen of 239 subjects aged 12 75 years from extended families suggested a high disease prevalence according to full-mouth pocket probing depths (PPDs), with more than 75% of subjects with one or more pockets of PPD > or =5 mm. A more detailed study was performed in 125 unrelated subjects > or =18 years, recording full-mouth PPDs and clinical attachment levels (CALs). The high prevalence of pocketing was confirmed and 90% of adults > or =35 years had at least one site with CAL > or =6 mm. However, extensive disease was restricted to a small minority, with only 10% of adults > or =35 years having 20% or more sites with CAL > or =6 mm. CONCLUSION: The study results highlight the importance of performing a detailed examination and appropriate analysis. In both studies, tooth retention was high (mean number of teeth recorded was 26.4 and 28.0 respectively), smoking unusual, and families large and localised to the village. This community thus affords several advantages over populations in developed countries when considering familial studies of adult periodontitis. PMID- 11422589 TI - Evaluation of the effect of tooth vitality on regenerative outcomes in infrabony defects. AB - BACKGROUND, AIMS: This investigation was designed to evaluate the null hypothesis of no differences in GTR outcomes in intrabony defects at vital and successfully root-canal-treated teeth. METHOD: 208 consecutive patients with one intrabony defect each were enrolled. Based on tooth vitality, the treated population was divided at baseline into 2 groups: one with 41 non-vital teeth and the other with 167 vital teeth. The 2 groups were similar in terms of patient and defect characteristics. RESULTS: A slight unbalance in terms of depth of the intrabony component was observed in the non-vital group compared to the vital group (6.9+/ 2.1 mm versus 6.2+/-2.3 mm, p=0.08). All defects were treated with GTR therapy. At 1 year, the non-vital and the vital groups showed a clinical attachment level (CAL) gain of 4.9+/-2.2 mm and of 4.2+/-2 mm, respectively. The difference was statistically significant (p=0.03). To correct for the baseline unbalance in defect depth, data were expressed as a % of clinical attachment level gains with respect to the original intrabony depth of the defect. % CAL gains were 72.8+/ 42.2% and 73+/-26.4% for vital and non-vital teeth, respectively: the difference was not statistically significant (p=0.48). Average residual pocket depths were 2.8+/-1 mm in the vital and 2.8+/-0.9 mm in the non-vital group. Tooth vitality was assessed at baseline, at 1-year and at follow-up (5.4+/-2.8 years after surgery): all teeth vital at baseline were still vital at follow-up with the exception of 2 teeth that received endodontic treatment for reconstructive reasons and for caries. At follow-up visit, the difference in CAL with respect to 1-year measurements was -0.9+/-0.8 mm in the vital group and -0.7+/-0.8 mm in the non-vital group, indicating stability of the regenerated attachment at the majority of sites. CONCLUSIONS: Data from this study demonstrate that root canal treatment does not negatively affect the healing response of deep intrabony defects treated with GTR therapy; furthermore GTR therapy in deep intrabony defects does not negatively influence tooth vitality. PMID- 11422590 TI - Tobacco smoking and periodontal hemorrhagic responsiveness. AB - OBJECTIVE: The influence of smoking on the periodontal hemorrhagic responsiveness was investigated in 2 different populations, one exclusively consisting of patients with established periodontal disease, the other of dental hospital admissions in general. MATERIAL AND METHODS: The hemorrhagic responsiveness of the patient was clinically found from "bleeding on gentle probing" expressed as the relative frequency of bleeding sites (%). In the periodontal patient population, gingival bleeding was assessed by 1 examiner with known probing force, whereas in the dental hospital admissions population assessments were done by a great number of non-calibrated examiners with unknown probing force. RESULTS: Smokers exhibited a significantly lower hemorrhagic responsiveness than non-smokers. This held principally true for both populations but the effect was much more clearly detectable in periodontal patients than in dental hospital patients in general. A dose-response effect was typically evident in the periodontal patient population. Accounting for the periodontal disease severity, however, the effect of smoking became clearly detectable also in the general patient population. CONCLUSION: Tobacco smoking is associated with a clinically suppressed hemorrhagic responsiveness of the periodontium. PMID- 11422591 TI - Mutans streptococci in subgingival plaque of treated and untreated patients with periodontitis. AB - BACKGROUND: The etiology of root caries is thought to be identical to coronal caries, though root caries seem to be more complicated because of the higher susceptibility of exposed roots (dentin) by periodontal therapy to demineralization than intact enamel. This implies that mutans streptococci are the most likely pathogens in the development of root caries. Although it is known that both the numbers of mutans streptococci and the frequency of isolation in root caries lesions are negatively correlated with the distance from the gingival margin, the subgingival sulcus has not been considered a possible habitat for mutans streptococci. However, subgingival occurrence of mutans streptococci in both untreated and treated periodontal patients has not been documented well in the literature. OBJECTIVE: To investigate the presence and levels of mutans streptococci in the subgingival plaque of patients (n=154) in different stages of periodontal therapy. The subgingival sulcus may be a possible habitat for mutans streptococci. This localisation of mutans streptococci may be of importance in the development of root caries after periodontal surgery. MATERIALS AND METHODS: In this cross-sectional study, subgingival plaque samples from 154 consecutive adult periodontitis patients were tested for presence and levels of mutans streptococci and putative periodontal pathogens by anaerobic cultures. These patients were divided into 4 groups based on their stage of periodontal treatment: (1) untreated patients; (2) patients after initial periodontal therapy only; (3) patients in the maintenance phase who not underwent periodontal surgery; (4) patients after periodontal surgery. RESULTS: The prevalence of mutans streptococci in the 4 study groups varied from 82% in untreated patients to 94% in maintenance patients. The mean proportion of mutans streptococci was 6.65% in maintenance patients versus 1.86% in untreated patients (p=0.005) and 2.51% in patients after scaling and root planing (p=0.041). CONCLUSIONS: The subgingival area is a microbial habitat for mutans streptococci that may be of importance in the development of root caries in periodontitis patients. PMID- 11422592 TI - Orofacial granulomatosis with gingival onset. AB - BACKGROUND, AIMS: Orofacial granulomatosis (OFG) is a descriptive term used for granulomatous disorders of the face and oral cavity that may occur for a variety of reasons, some of which result in significant morbidity and mortality. Although rarely, a granular enlargements of the gingiva may be the first clinical manifestation of OFG, preceding other local or systemic manifestations. METHOD: We will report a case of OFG that showed an atypical and monosymptomatic onset with a generalized hyperplastic gingivitis that preceded other facial and mucosal features by several weeks. RESULT: Considering the variable clinical onset of OFG and its apparent increase in incidence, we emphasize that in some cases, the periodontologist, as first consulted health care professional, plays an important role to detect this disorder. Early diagnosis of OFG is a crucial step to prevent and cure its unsightly sequelae and sometimes avoid progression of systemic potentially life-threatening OFG-related diseases. CONCLUSION: Thus, when none of the common causes of gingival enlargement can be detected, OFG diagnosis should be suspected. PMID- 11422593 TI - Comparative antimicrobial activities of antiseptic mouthrinses against isogenic planktonic and biofilm forms of Actinobacillus actinomycetemcomitans. AB - BACKGROUND: Bacteria contained in biofilms have been shown to have a decreased susceptibility to antimicrobial agents compared to those in planktonic form. Thus, in vitro biofilm models have been developed for screening oral antimicrobial formulations in an effort to produce findings more predictive of clinical activity. This study compared the antimicrobial activity of three mouthrinse formulations when tested against isogenic strains of Actinobacillus actinomycetemcomitans (Aa), one of which was a clinical isolate which forms tenacious biofilms in vitro and the other of which was a spontaneous variant which always grows planktonically. METHOD: Biofilm-forming Aa strains CU1000 and NJ4300, obtained as clinical isolates, and their respective spontaneous planktonic variants, CU1060 and NJ4350, were grown under standard laboratory conditions and exposed for 15 s to either a negative control (phosphate buffered saline [PBS]), an essential-oil containing mouthrinse (Listerine Antiseptic [LA]), an amine fluoride/stannous fluoride-containing mouthrinse (Meridol [M]), or a triclosan and PVM/MA copolymer-containing mouthrinse (Plax [P]). The cells were then washed, serially diluted, plated, and incubated for enumeration of viable bacteria. Colony-forming units (CFU)/ml were log10 transformed and the mouthrinse groups were compared to the PBS group using analysis of variance. RESULTS: All 3 mouthrinses produced statisically significant 99.99% reductions (p< or =0.0001) in both planktonic strains compared to the PBS control. Effects on the biofilm forms of the organisms were more variable. Exposure to LA produced statistically significant (p< or =0.0001) reductions in strains CU1000 and NJ4300 of 98.20% and 96.47%, respectively, compared to PBS. M and P produced much smaller reductions which were not statistically significant. CONCLUSIONS: The results of this study, in which antimicrobial mouthrinses were tested against biofilm-forming and planktonic strains of the same organism, provide a clear demonstration of the resistance to antimicrobial agents conferred by biofilm formation and provide additional support for employing tests using biofilms to more accurately assess the relative activities of antiplaque agents in vitro. PMID- 11422594 TI - Capnocytophaga granulosa and Capnocytophaga haemolytica: novel species in subgingival plaque. AB - BACKGROUND: The oral cavity accommodates one of the most diverse microfloras in the human body. Knowledge of this microflora, and of the periodontal microflora in particular, proves crucial towards an understanding of the bacterial-host interactions which lead to the development of infectious inflammatory periodontal diseases. Capnocytophaga species have been implicated as putative periodontal pathogens. To date, only 3 members of this genus (C. gingivalis, C. ochracea and C. sputigena) have been isolated from subgingival plaque. AIM: This communication reports the isolation of 2 recently-speciated strains, namely C. granulosa and C. haemolytica, from subgingival plaque collected from adult periodontitis patients. MATERIAL AND METHODS: Subgingival plaque was collected from 29 patients with chronic adult periodontitis. Plaque samples were inoculated onto fastidious anaerobe agar and incubated anaerobically for 5 days. Routine identification of clinical isolates was performed by 16S rRNA PCR-RFLP analysis, using Cfo I as restriction enzyme and corroborated by 16S rRNA gene sequencing. RESULTS: 16 of 29 patients (55%) tested positive for either C. granulosa and or C. haemolytica. A total of 70 isolates (63 C. granulosa and 7 C. haemolytica) were cultivated from subgingival plaque. 15 (51%) patients tested positive for C. granulosa, and 3 (10%) patients tested positive for C. haemolytica. CONCLUSION: This is the 1st report which recounts the presence of C. granulosa and C. haemolytica in subgingival plaque. Further research is required to establish the relative proportions of these species subgingivally in health and disease. PMID- 11422595 TI - Cyclosporin A-induced gingival overgrowth is unrelated to allograft function in renal transplant recipients. AB - BACKGROUND: Severe gingival hyperplasia (GH) is one of the most frequent side effects associated with the prescription of Cyclosporine-A (CsA). AIM: This study statistically modeled the medical and dental risk factors for the development of GH following CsA administration to determine whether renal function post transplantation was related to the incidence or extent of GH in 236 consecutive renal transplant patients. METHOD: All patients were at least 6 months post transplant and medicated with both traditional oral CsA (n=220 individuals) and the new microemulsion form CsA-Me (n=229 individuals). Patients had either received CsA alone (n=45 individuals) or cyclosporine and nifedipine (n=191 individuals). Gingival overgrowth was assessed and computerized data, available for all patients included; pre- and post-transplant medical history and post transplant renal function, i.e., serum creatinine levels, documented rejection episodes and glomerular filtration rates (GFR). These data together with CsA serum levels and last-recorded dose of CsA, CsA-Me, nifedipine, azathioprine and prednisolone, were analysed by multivariate regression analysis using SPSS. RESULTS: The extent and severity of hyperplasia was significantly correlated with the dosage and serum level of CsA at 3, 6 and 12 months post-transplantation; last recorded dosage, however (p<0.0001), was the most accurate predictor of hyperplasia. Gingivitis (p<0.0001) and plaque (p<0.002), were associated with hyperplasia. Duration of renal replacement therapy, age at transplantation, post transplant interval serum creatinine levels and documented rejection episodes were unrelated with the extent and severity of GH. Of all the renal variables only the correlation of GFR with last recorded doses of CsA and CsA-Me, approached significance; this was then considered for inclusion in the model. CONCLUSION: In a multiple regression analysis including GFR, however, only last CsA (and CsA-Me) doses and gingivitis score were selected for inclusion in the final model. These data demonstrate that inter-patient variation in the extent and severity of GH and renal function post-transplantation are unrelated and are mediated independently. PMID- 11422596 TI - Elevated blood glucose levels in patients with severe periodontal disease. AB - BACKGROUND, AIMS: The role of diabetes mellitus in various forms of periodontal disease was investigated intensively in the past; some studies have also indicated an impact of periodontal inflammation on diabetic balance. The purpose of this study was to evaluate the reciprocal relationship between abnormal serum glucose levels and elevated CPITN scores in non-diabetic patients. MATERIAL AND METHODS: 10,590 subjects were analyzed for normal and abnormal (higher than 120 mg/dl) serum glucose levels, their periodontal status reflected as CPITN was correlated with their glucose level category. RESULTS: The results of the present study reflect a significant association between elevated blood glucose levels (higher than 120 mg/dl) and CPITN scores of 4.5 with an odds ratio of 2.46. CONCLUSION: A strong association exists between abnormal serum glucose level and periodontal disease as manifested by CPITN score. PMID- 11422597 TI - The noncompliance blame game. PMID- 11422598 TI - Itraconazole oral solution and intravenous formulations: a review of pharmacokinetics and pharmacodynamics. AB - Itraconazole is a triazole antifungal agent with a broad spectrum of activity. It is well tolerated and highly efficacious, particularly because its main metabolite, hydroxy-itraconazole, also has considerable antifungal activity. Two new formulations of itraconazole, an oral solution and an intravenous formulation, have recently been developed, which combine lipophilic itraconazole with cyclodextrin. These formulations have improved the solubility of itraconazole, leading to enhanced absorption and bioavailability compared with the original capsule formulation, without having an impact on the tolerability profile of itraconazole. The oral solution and intravenous formulations of itraconazole produce consistent plasma concentrations and are ideal for the treatment of systemic fungal infections in a wide range of patient populations. The additional flexibility offered by the different routes of administration means that itraconazole treatment can be specifically tailored for use in all patients, including children and those requiring intensive care. PMID- 11422599 TI - Identification of the precipitate in alkalinized solutions of mepivacaine and bupivacaine at 37 degrees C. AB - Commercial and control solutions of bupivacaine (0(.)75%) and mepivacaine (1(.)5%) were alkalinized with bicarbonate until cloudy at room temperature. The solutions were heated to 37 degrees C for 2(.)5 h. The precipitates were filtered, lyophylized and analysed by fast atom bombardment mass spectrometry. Analysis showed the precipitates to be predominantly the free base of the local anaesthetic. The precipitate of the commercial bupivacaine solution also contained a small amount of the hydrochloride salt. The mepivacaine control crystals contained an unknown at molecular weight 528, which may represent a dimer of the free base and hydrochloride salt. PMID- 11422600 TI - Effect of temperature on serum protein binding characteristics of phenytoin in monotherapy paediatric patients with epilepsy. AB - OBJECTIVES: To determine the effects of temperature on binding characteristics of phenytoin to serum proteins in paediatric patients with epilepsy. METHOD: Serum samples examined in the study were obtained from 41 paediatric patients (23 male, 18 female) with epilepsy on phenytoin monotherapy. Their age ranged from 1 to 15 years (mean +/- SD, 10;2 +/- 4;0 years). Protein binding of phenytoin was evaluated by ultrafiltration under current laboratory routine conditions (25 +/- 3 degrees C) or at a temperature of 37 degrees C. The in vivo binding parameters of phenytoin to serum proteins were determined using a binding equation derived from the Scatchard equation for a one-site binding model. RESULTS: Significant differences were observed in serum concentrations of unbound phenytoin at the two temperatures (P < 0;05). The mean association constant L/micromol (K) of phenytoin to serum proteins is 0.016 L/micromol at 25 +/- 3 degrees C and 0;009 L/micromol at 37 degrees C, while mean total concentration of binding sites (n(Pt)) seems to be similar between the two temperatures (682 micromol/L for 25 +/- 3 degrees C and 746 micromol/L for 37 degrees C). Significant differences were observed in binding characteristics of phenytoin to serum proteins for the different temperature conditions of ultrafiltration (P < 0;05). CONCLUSION: Our study confirms that binding affinity for phenytoin-serum protein interaction is approximately 44% lower at 37 degrees C than at 25 +/- 3 degrees C and consequently, binding potential (K.n(Pt)) is approximately 38% lower at 37 degrees C than at 25 +/- 3 degrees C. PMID- 11422601 TI - Cerebral embolism and hormone replacement therapy. AB - Few studies have focused on the relationship between hormone replacement therapy (HRT) for postmenopausal women or those with breast cancer and the occurrence of cerebral embolism. Results are conflicting as to whether there is a link between the two. We describe three patients who experienced cerebral embolism during HRT. A 73-year-old woman had a transient ischemic attack (TIA) 6 years prior to the present admission. She then took HRT oestrogen plus medroxyprogesterone acetate for about 6 years. The HRT had been prescribed by a gynaecologist for amelioration of postmenopausal symptoms. Six years after beginning HRT, she experienced sudden onset left hemiparesis due to cerebral embolism. Two other patients had been taking HRT for breast cancers. One, a 47-year-old woman, had taken medroxyprogesterone acetate for more than one year, for recurrence of breast cancer. She had developed sudden complete left hemiparesis due to an embolism at the carotid bifurcation. The other patient, a 72-year-old woman who was taking tamoxifen citrate for prevention of breast cancer relapse, experienced cerebral embolism just 2 months after beginning tamoxifen. The risk of cerebral embolism in those on HRT should be emphasized, along with the beneficial effects in terms of postmenopausal symptoms and prevention of breast cancer recurrence. PMID- 11422602 TI - Influence of seven beverages on salicylate disposition in humans. AB - OBJECTIVE: Aspirin administered orally is one of most widely self-prescribed drugs to treat headaches or other pains. The aim of this study was to evaluate whether the influence of different beverages may be used to help in the ingestion of an aspirin tablet on the pharmacokinetic parameters of this drug. METHOD: This study was undertaken in five healthy volunteers. Seven beverages were tested: water, tea, coffee, orange juice, milk, beer and 40 degrees distilled alcohol. After plasma extraction, aspirin and salicylic acid were measured by HPLC with UV detection. The main pharmacokinetic parameters were determined by the compartimental method and drug disposition profiles by the Wagner-Nelson modified method. RESULTS: Elimination was not modified by any of the beverages but absorption was affected. Two opposing effects were observed: 40 degrees alcohol seemed to increase AUC and Cmax. Milk and beer seemed to decrease these parameters. With 40 degrees alcohol and tea, the amount absorbed and the disposition rate were higher. For milk and orange juice, the amount absorbed was lower and the disposition rate was unaffected. For beer, both the amount absorbed and the disposition rate increased. For coffee, both the amount absorbed and disposition rate were not significantly modified. CONCLUSION: The bioavailability of salicylates on the healthy volunteers in this study was significantly affected by concomitant administration of 40 degrees alcohol (spirit), beer and milk. The beverages seem to interfere with aspirin absorption and the drug disposition profile was modified. PMID- 11422603 TI - Antibiotic prescribing by single handed general practitioners: secondary analysis of data. AB - AIM: To determine the contribution of various doctor and patient factors on the frequency of antibiotic prescribing. METHODS: Secondary analyses of data on 155 single handed general practitioners. RESULTS: Three variables explained 25% of variation in antibiotic prescribing. Doctors qualified from the Indian subcontinent issued more antibiotics than U.K.-qualified doctors. Patients from the non-manual social class were issued fewer antibiotics than those from the manual class and the most deprived patients received significantly more antibiotics. CONCLUSION: Very little of prescribing of antibiotics by doctors is explained by these doctor-patient factors. Prescribing is a complex process and the search for factors must continue in order to address the rising antibiotic resistance. PMID- 11422604 TI - A leucine(7)-to-proline(7) polymorphism in the signal peptide of neuropeptide Y was not identified in the Japanese population. AB - OBJECTIVE: A Leu(7)-to-Pro(7) polymorphism exists in the signal peptide part of prepro-neuropeptide Y (NPY) and this polymorphism is associated with high serum cholesterol and LDL cholesterol levels in both Finnish and Dutch people. There was a significant difference in the allelic frequencies among populations with different genetic backgrounds. Therefore, the present study was designed to test whether Japanese people had the Leu(7)-to-Pro(7) polymorphism in the NPY signal peptide. METHODS: A total of 102 healthy Japanese volunteers supplied blood samples, from which leucocyte DNA was isolated. Primer pairs for the polymerase chain reaction (PCR) were made according to Karvonen et al. Direct sequencing of the PCR product was carried out by the dye terminator method using a capillary electrophoresis sequencer. RESULTS: All 102 young healthy Japanese subjects showed Leu(7)/Leu(7) but not Leu(7)/Pro(7) or Pro(7)/Pro(7) in the NPY signal peptide. CONCLUSION: The incidence of the Leu(7)-to-Pro(7) polymorphism in the signal peptide of NPY was extremely low in Japanese people, suggesting that the polymorphism producing Pro(7) is not one of the genetic determinant factors affecting serum cholesterol levels in Japanese people. PMID- 11422605 TI - Heterogeneity of the CYP2D6 gene among Malays in Malaysia. AB - BACKGROUND: Although Malays shared an origin with Chinese, their evolution saw substantial divergences. Phenotyping studies suggested that they differed in CYP2D6 polymorphism, with higher PM prevalence but lesser right-shift for debrisoquine MRs. OBJECTIVE: To study the genotype distribution of CYP2D6 among the Malays in Malaysia. METHOD: We obtained DNA from 107 Malays and used PCR to determine common CYP2D6 alleles. RESULT: CYP2D6*1 occurred at a frequency of 36.0%, duplicated gene, 0.93%, CYP2D6*4, 2.8%, CYP2D6*5, 5.1%, CYP2D6*9, 3.3%, CYP2D6*10, 49.5% and CYP2D6*17, 0.5%. The findings of CYP2D6*17 and CYP2D6*9 were novel for Asia. The frequency for CYP2D6*10 was lower than in other Asian races. The most frequent genotypes were CYP2D6*1/*10 at 39.3%. Two subjects had genotypes that predicted PM phenotype, 35% showed genotypes that predicted intermediate metabolizers and one subject had a genotype that predicted ultra rapid metabolism. CONCLUSION: The genetic polymorphism of CYP2D6 in Malays is different from Chinese and Far Eastern races. They may be intermediate between East Asians and Caucasians in CYP2D6 activity. Further study in relation to the evolution of races and disease prevalence may help to identify the contributions of the polymorphism in alleged susceptibility to diseases apart from delineating its contributions to ethnic differences in the pharmacology of CYP2D6 drugs. PMID- 11422606 TI - Population pharmacokinetic modelling of carbamazepine by using the iterative Bayesian (IT2B) and the nonparametric EM (NPEM) algorithms: implications for dosage. AB - OBJECTIVE: To estimate individual and population postinduction pharmacokinetics of carbamazepine (CBZ) in epileptic adult and paediatric patients who received chronic CBZ monotherapy. METHODS: We have used the USC*PACK collection of PC programs for the estimations. The preinduction CBZ metabolism was also estimated in 16 volunteers after a single dose of CBZ (200 mg). We used a linear one compartmental model with oral absorption and found the pharmacokinetic parameter values of CBZ behaviour to be in good agreement with those reported earlier. RESULTS: Serum CBZ concentrations correlated poorly with daily doses in both the adult and child populations. Because of the diversity within the population, use of the mean population model without knowledge of an individual patient's pharmacokinetic characteristics gives poor prediction. In contrast, the individual Bayesian posterior models gave good prediction for all subjects in the population, due to the removal of the interindividual variability. CONCLUSION: This approach permits one to individualize drug therapy for patients even when only sparse therapeutic drug monitoring (TDM) data are available. Future individual CBZ serum level predictions were acceptable from a clinical point of view (mean absolute error = 13.2 +/- 9.7%). The optimal sampling strategy approach helped to design an optimal cost-effective TDM protocol for CBZ therapy management. PMID- 11422607 TI - Severe toxic hepatitis associated with amoxycillin and clavulanic acid. AB - Toxic hepatitis secondary to amoxycillin-clavulanic acid is an infrequent clinical picture. Most of the cases are reported to have a benign course. We report two cases of severe hepatic failure following amoxycillin-clavulanic acid use. One of the cases had cholestatic features primarily, and the other had hepatocellular injury prominently. The first case had also findings of trombotic trombositic purpura and had a fatal course. PMID- 11422608 TI - Role of coagulation in the natural history and treatment of portal vein thrombosis. PMID- 11422609 TI - Concurrent hepatitis B and C virus infection: Is hepatitis C virus stronger? PMID- 11422610 TI - Lipid peroxidation in hepatic steatosis in humans is associated with hepatic fibrosis and occurs predominately in acinar zone 3. AB - BACKGROUND AND AIMS: Hepatic steatosis has been shown to be associated with lipid peroxidation and hepatic fibrosis in a variety of liver diseases including non alcoholic fatty liver disease. However, the lobular distribution of lipid peroxidation associated with hepatic steatosis, and the influence of hepatic iron stores on this are unknown. The aim of this study was to assess the distribution of lipid peroxidation in association with these factors, and the relationship of this to the fibrogenic cascade. METHODS: Liver biopsies from 39 patients with varying degrees of hepatic steatosis were assessed for evidence of lipid peroxidation (malondialdehyde adducts), hepatic iron, inflammation, fibrosis, hepatic stellate cell activation (alpha-smooth muscle actin and TGF-beta expression) and collagen type I synthesis (procollagen alpha1 (I) mRNA). RESULTS: Lipid peroxidation occurred in and adjacent to fat-laden hepatocytes and was maximal in acinar zone 3. Fibrosis was associated with steatosis (P < 0.04), lipid peroxidation (P < 0.05) and hepatic iron stores (P < 0.02). Multivariate logistic regression analysis confirmed the association between steatosis and lipid peroxidation within zone 3 hepatocytes (P < 0.05), while for hepatic iron, lipid peroxidation was seen within sinusoidal cells (P < 0.05), particularly in zone 1 (P < 0.02). Steatosis was also associated with acinar inflammation (P < 0.005). alpha-Smooth muscle actin expression was present in association with both lipid peroxidation and fibrosis. Although the effects of steatosis and iron on lipid peroxidation and fibrosis were additive, there was no evidence of a specific synergistic interaction between them. CONCLUSIONS: These observations support a model where steatosis exerts an effect on fibrosis through lipid peroxidation, particularly in zone 3 hepatocytes. PMID- 11422611 TI - Intestinal dysfunction in liver cirrhosis: Its role in spontaneous bacterial peritonitis. AB - Spontaneous bacterial peritonitis is a common illness in patients with cirrhosis and ascites that occurs without any apparent focus of infection. Bacterial translocation plays an important role in spontaneous bacterial peritonitis and it is evident from a variety of studies that the gut is a major source of this bacteria. Gut motility alterations, along with bacterial overgrowth and changes in intestinal permeability, probably play a role in this bacterial translocation. The present review looks at the role of the intestine in spontaneous bacterial peritonitis induced by liver cirrhosis and the factors influencing bacterial translocation in this disease. PMID- 11422613 TI - Influence of climatic factors in the incidence of upper gastrointestinal bleeding. AB - BACKGROUND AND AIM: Previous reports have indicated seasonal fluctuations in the incidence of peptic ulcer activity, but the reasons for the seasonal pattern are not clear. We assessed the seasonal incidence of hematemesis caused by peptic ulcers or gastroesophageal varices, and the correlations between those and climatic factors. METHODS: We examined the number of cases of upper gastrointestinal (GI) bleeding caused by gastric ulcer (GU), duodenal ulcer (DU), or gastroesophageal varices (varix) diagnosed by urgent endoscopies between 1 January 1996 and 31 December 1999 in our hospital (Tokyo Metropolitan Bokutou Hospital). We evaluated the monthly and seasonal incidence of them and investigated correlations among the incidence and climatic factors. RESULTS: Four hundred and forty-one patients participated in this study, including 275 patients with GU (62.4%), 51 (11.6%) with DU, and 115 (26.0%) with varix. The number of cases of hematemesis caused by GU showed significant monthly and seasonal fluctuations (P = 0.0002, P = 0.0018): it decreased in summer and increased in autumn-winter. Moreover, there were inverse relations between the monthly number of cases of hematemesis caused by GU and the mean temperature (P = 0.0016) and vapor pressure (P = 0.0013), and a parallel relation to the mean atmospheric pressure (P = 0.0057). In contrast, the number of cases of hematemesis caused by DU and varices did not show any monthly or seasonal fluctuations. CONCLUSIONS: We found that the incidence of hematemesis because of GU had an inverse relationship to temperature and vapor pressure, and had a parallel relation to atmospheric pressure. Therefore, climatic factors may play an important role in hemorrhage from GU. PMID- 11422612 TI - Strategy for retreatment of therapeutic failure of eradication of Helicobacter pylori infection. AB - BACKGROUND AND AIM: A proton pump inhibitor (PPI)-based triple therapy consisting of a PPI, amoxicillin (A) and clarithromycin (C) or metronidazole (M) provides an eradication rate ranging from 80 to 90%. However, there have been few controlled studies with regard to the most effective regimen to re-treat patients after failure of the first-line therapy. Accordingly, we retrospectively reviewed our experiences and compared regimens with different combinations of antimicrobials to determine the optimal retreatment regimen. METHODS: Out of 133 patients who had received second-line therapy after failure of first-line PPI/AC therapy, we selected, for review, patients who took the prescribed drugs for first-line therapy equal to, or more than 80%. As a result, data on 114 patients (83 males and 31 females; mean age 49.1 +/- 13.0 years; peptic ulcer n = 89; non-ulcer dyspepsia, n = 25) were eligible for evaluation. They had either repeated the PPI/AC regimen (n = 34; 5-14 days), or had been administered the PPI/AM regimen (n = 80; 10 days). The cure rates of the two regimens were compared. RESULTS: The eradication rates for second-line therapy with the PPI/AC regimen versus the PPI/AM regimen were 52.9% (95% CI, 35-70) versus 81.3% (95% CI, 71-89) by intention-to-treat analysis (P < 0.01), and 62.1% (95% CI, 42-79) versus 91.4% (95% CI, 81-97) by per-protocol analysis (P < 0.01). CONCLUSION: The eradication rate for the PPI/AM retreatment regimen was significantly higher than for the repeated PPI/AC regimen, suggesting that a 10-day PPI/AM regimen can be recommended as a retreatment regimen for patients who had first-line eradication therapy by PPI/AC regimens. PMID- 11422614 TI - Gastrointestinal transit and anorectal manometry in children with colonic substance P deficiency. AB - BACKGROUND AND AIMS: Severe intractable constipation in children may be associated with a reduction of substance P (SP)- containing fibers in colonic circular muscle. The aim of this study was to characterize gastrointestinal transit (GIT), anorectal manometry (ARM) and electromyographic (EMG) changes in these children. METHODS: Seromuscular laparoscopic biopsies of the colon were obtained from 35 children with severe constipation. Immunofluorescent staining for SP and vasoactive intestinal peptide (VIP) were then performed on these specimens. The cohort of patients studied included a SP-deficient group (SPD, n = 25) who had reduced numbers of SP-immunoreactive nerve fibers. The other group consisted of patients with normal staining for both SP and VIP (SPN, n = 10). Gastrointestinal transit studies (gastric emptying, orocecal and colonic transit) suitable for analysis were available for 17 patients (SPD, n = 9 and SPN, n = 8). The colon was divided into segments and radioactivity counts in each segment were expressed as a percentage of the total colonic count at each time point (6, 24, 32 and 48 h). The geometric center (GC), ARM, EMG, clinical and demographic data characteristics of both groups of patients were compared. RESULTS: There were no differences in demographic data, gastric emptying, orocecal transit or geometric center of transit in the colon between the two patient groups. The ARM and EMG studies suggested that the SPN group have a higher mean threshold volume of balloon distension required to initiate a rectoanal inhibitory reflex, and a higher incidence of anismus; however, this did not reach statistical significance. CONCLUSIONS: These data suggest a trend that the SPN patients have a greater problem with obstructive defecation and abnormal rectal sensation than those with SPD. We were unable to confirm any defect in colonic transit in the SPD patients compared with the SPN group. PMID- 11422615 TI - Metabolic genotypes and risk for colorectal cancer. AB - BACKGROUND: Inherited polymorphisms that influence carcinogen metabolism or the composition of bile may influence the risk for the development of colorectal cancer. METHODS: The frequency of polymorphisms involving N-acetyltransferase 1 (NAT1), NAT2, cytochrome P450 1A1 (CYP1A1), CYP2D6, CYP2E1, glutathione S transferase M1 (GSTM1), GSTT1 and apolipoprotein E were compared in 219 white adults with sporadic colorectal cancer and 200 white controls attending for blood donation at a blood bank. Polymorphisms were determined by using restriction fragment length polymorphism (RFLP) after amplification of genomic DNA by polymerase chain reaction (PCR). Data were analyzed by using standard statistical methods for a case- control study, and reported as odds ratios (OR) with 95% confidence intervals (CI). RESULTS: None of the genotypes, either alone or in combination, showed a strong association with colorectal cancer. Inheritance of the GSTT1 null genotype conferred a twofold risk of cancer that was statistically significant with crude data (OR 2.18; 95% CI 1.38-3.43), but not after adjustment for age (OR 1.91; 95% CI 0.99-3.70). There was also a trend towards a lower risk for proximal (right-sided) cancers in patients with apolipoprotein epsilon4 (OR 0.64; 95% CI 0.31-1.33). CONCLUSION: No strong associations have been found between metabolic genotypes and colorectal cancer risk in Australia. Large studies will be required to confirm weak associations and to establish relationships between cancer risk, metabolic genotypes and exposure to dietary or other environmental carcinogens. PMID- 11422616 TI - Influence of hepatitis C virus on the profiles of patients with chronic hepatitis B virus infection. AB - BACKGROUND: Hepatitis B virus (HBV) and hepatitis C virus (HCV) are the most common causes of chronic liver disease, cirrhosis and hepatocellular carcinoma. The influence of HCV infection on the clinicopathological and virological profiles of chronic HBV infection was investigated. METHODS: A total of 100 chronic HBV carriers with histopathological diagnoses by liver biopsy were studied. Hepatitis B e antigen (HBeAg) and anti-HCV antibody were tested. Serum HCV-RNA was detected by using a nested reverse transcription-PCR assay. A branched DNA (bDNA) assay was used to detect HBV-DNA and quantitate the serum levels. RESULTS: Eighteen (18%) of 100 patients were positive for anti-HCV and HCV-RNA. Patients with concurrent HCV and HBV infection were significantly older than those without HCV infection (P < 0.05). The positive rates of HBeAg and HBV DNA as well as the serum levels of HBV-DNA in patients with concurrent HCV and HBV infection were significantly lower than those without concurrent HCV and HBV infection (P < 0.01, P < 0.05, and P < 0.001, respectively). By using multivariate analysis, the factors of seroconversion of HBeAg and decreasing level of HBV-DNA were significantly correlated to concurrent HCV and HBV infection in chronic HBV carriers. The factors of increasing age and concurrent HCV and HBV infection were significantly correlated to seroconversion of HBeAg. CONCLUSIONS: The concurrent HCV and HBV infection in chronic HBV carriers might result in a suppression of HBV replication that presented with a lower level of serum HBV-DNA and HBeAg seroconversion. Nevertheless, neither more obvious increase in biochemical parameters nor histopathological progression to more advanced liver diseases was observed. PMID- 11422617 TI - Coagulation profile and platelet function in patients with extrahepatic portal vein obstruction and non-cirrhotic portal fibrosis. AB - BACKGROUND AND AIMS: Coagulation disorders commonly develop in patients with cirrhosis of the liver. They have also been reported in patients with non cirrhotic portal fibrosis (NCPF) and extra-hepatic portal venous obstruction (EHPVO); the two conditions with portal hypertension and near-normal liver functions. The spectrum and prevalence of coagulation abnormalities and their association with the pathogenesis of these diseases and with hypersplenism was prospectively studied. METHODS: Eighteen EHPVO patients that included an equal number of NCPF patients and 20 healthy controls were prospectively studied. The coagulation parameters assessed included: international normalized ratio, partial thromboplastin time, and fibrinogen and fibrinogen degradation products. Platelet aggregation and malondialdehyde levels were measured. RESULTS: Both EHPVO (83%) and NCPF (78%) patients had a significantly prolonged international normalized ratio and a decrease in fibrinogen and platelet aggregation. The EHPVO patients had a significant prolongation in partial thromboplastin time (67% patients), with increased levels of fibrinogen degradation product levels occurring in all patients; these were normal in NCPF patients. Platelet malondialdehyde levels were normal in both groups. Hypersplenism was present in four EHPVO and seven NCPF patients. It did not significantly influence the coagulation profile in either NCPF or EHPVO patients. CONCLUSIONS: Coagulation anomalies are common and significant in both NCPF and EHPVO patients, suggestive of a mild disseminated intravascular coagulation disorder. These imbalances could be caused by chronic subclinical endotoxemia and cytokine activation after the initial portal thromboembolic event. The persistence of these abnormalities in adolescent patients indicates an ongoing coagulation derangement. PMID- 11422618 TI - Relationship among gastric motility, autonomic activity, and portal hemodynamics in patients with liver cirrhosis. AB - BACKGROUND AND AIMS: We examined the effects of the autonomic nervous function and the volume of portal blood flow to clarify the mechanism of the abnormal gastric motility in patients with liver cirrhosis. METHODS: Heart rate variability, electrogastrogram (EGG), and volume of portal blood flow were measured before and after a meal in 27 patients with liver cirrhosis (LC group) and in 20 normal subjects (N group). Autonomic nervous function was evaluated by using spectral analysis of heart rate variability. We used the cine phase contrast (PC) method, using magnetic resonance imaging (MRI) to measure the portal flow, while the peak frequency and spectral power of the EGG were measured at pre- and postprandial change. RESULTS: The ratio of low frequency power to high frequency power (LF/HF) was significantly higher, and the HF power was significantly lower in the LC group than in the N group both before and after a meal. In both groups, the electrogastrographic peak power ratio before and after a meal showed a positive correlation with the HF ratio, and an inverse correlation with the LF/HF ratio. In addition, portal blood flow volume was significantly decreased in the LC group than in the N group. However, the increased rate of portal blood flow after a meal correlated positively with the increased rate of electrogastrographic peak power. Moreover, gastric motility was positively correlated with esophageal varices and coma scale with the use of multivariate analysis. CONCLUSIONS: Parasympathetic hypofunction, sympathetic hyperfunction and portal hemodynamics were closely related with gastric motility in cirrhotic patients. In addition, gastric motility was decreased, at least in part, by the ingestion of food in cirrhotic patients because of abnormalities in autonomic functions and portal blood flow following a meal. PMID- 11422619 TI - Radiation therapy in combination with transcatheter arterial chemoembolization for hepatocellular carcinoma with extensive portal vein involvement. AB - AIM: The aim of this study was to examine the effectiveness and toxicity of radiation therapy in combination with transcatheter arterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) with extensive portal vein tumor thrombus (PVTT). METHODS: The combined therapy was performed in 24 HCC patients with extensive PVTT. External radiation targeted for PVTT (50 Gy in 2 Gy fractions) was performed in combination with repetitive TACE for intralobar lesions using 30-60 mg epirubicin every 3 months, and associations of the following variables with the survival rate were evaluated: gender, age, viral etiology, Child's class, performance status, extrahepatic metastasis, size and number of HCC, and location of PVTT. RESULTS: The local response confined to PVTT was complete response (CR) in four patients, partial response (PR) in eight patients, no change (NC) in eight patients, and progressive disease (PD) in four patients. By using the stepwise Cox's regression analysis, only Child's class was associated with the survival rate. The survival rates after 1 and 2 years were 73 and 21% in Child's A, 10 and 0% in Child B or C, and 61 and 21% in patients in whom the local response was CR or PR, and 19 and 9% in those in whom the local response was NC or PD, respectively. By using the multiple logistic regression analysis, Child's class was the only factor associated with the local response (P = 0.006). CONCLUSIONS: The combined therapy is feasible and may be useful to reverse PVTT in patients with good hepatic function reserve. PMID- 11422620 TI - Profile of hepatocellular carcinoma in India: an insight into the possible etiologic associations. AB - BACKGROUND: Several etiologic factors including hepatitis viruses, alcohol and aflatoxin have been implicated in the pathogenesis of hepatocellular carcinoma (HCC). There is, however, limited information from the Indian subcontinent. METHODS: Seventy-four consecutive cases of HCC were studied. A detailed history, tests for hepatitis B virus (HBV; HBsAg, HBeAg, anti-HBe, IgG anti-HBc, anti-HBs and HBV-DNA), hepatitis C virus (HCV; anti-HCV and HCV-RNA) infection, liver histopathology and HBV-DNA integration by using Southern blot hybridization were studied. A p53 gene mutation was also studied by using PCR and single-strand conformation polymorphism. RESULTS: Hepatocellular carcinoma patients were predominantly males (mean age 49.5 +/- 14.0 years). Portal hypertension and cirrhosis were seen in 56 (76%) patients, more often (P < 0.05) in viral marker positive cases. Forty-five percent of patients had features of hepatic decompensation at presentation. Evidence of HBV infection was present in 53 (71%) patients. Twenty-six (49%) of these patients had either HBeAg + ve, HBV-DNA + ve (n = 12), or HBsAg - ve, HBV-DNA + ve (n = 14) forms of HBV infection. Hepatitis B virus DNA integration in the liver tissue was seen in 10 of 17 (59%) patients. Infection with HCV alone was detected in three (4%) and dual HBV and HCV infection in six (8%) patients. A majority (78.5%) of the chronic alcoholics had associated viral infection. The etiology of HCC remained undetermined in 15 (20%) patients. The p53 gene mutations were detected only in three of 21 (14%) liver tissues. Aflatoxin toxicity, oral contraceptive use or metabolic disorder were not seen. CONCLUSIONS: In India: (i) HBV infection is the predominant factor for the development of HCC, often related to mutant forms of HBV; (ii) a majority of the HCC patients have overt cirrhosis of the liver; and (iii) HCV and alcohol per se are uncommonly associated. PMID- 11422621 TI - Autoimmune liver disease in children. AB - BACKGROUND AND AIM: Autoimmune liver disease (AILD) in children progresses to cirrhosis and liver failure if not diagnosed and managed in time. We prospectively analyzed our patients with liver disease for autoimmune etiology and their outcome with treatment. METHODS: All patients with liver disease were evaluated with liver function tests, abdominal ultrasonography, endoscopy, liver biopsy, viral markers and investigations for Wilson's disease. Immunoglobin (Ig)M hepatitis A virus, hepatitis E virus (HEV) and IgM hepatitis B core antibody were tested if acute viral hepatitis was suspected. Antinuclear antibody (ANA), antismooth muscle antibody (SMA), and liver kidney microsomal antibody (anti-LKM 1) were done in all cases. Autoimmune liver disease was diagnosed when one or more autoantibodies tested positive (> 1:40), and no other etiology of liver disease was identified. We also applied criteria proposed by the International Autoimmune Hepatitis Group. Cases diagnosed to have AILD were treated with immunosuppressive drugs. RESULTS: Autoimmune liver disease constituted 3.9% (6/153; median age and duration of illness 8.5 years and 3 months, respectively) of chronic liver disease cases. Four patients had acute hepatitis-like presentation. Of the six cases, two each were ANA and SMA +; one was anti-LKM-1 +, and the other was positive for both SMA and anti-LKM-1. Three of the patients achieved remission with combination therapy of oral prednisolone (OP) and azathioprine (AZT), and one with only OP. The other two patients were not treated. Two of the patients in remission have been weaned off from immunosuppressive therapy, and one is in a withdrawal phase. Another patient, while in biochemical remission developed superimposed anicteric acute HEV infection. CONCLUSION: Although AILD is uncommon in children, its search is rewarding, as remission is achieved with immunosuppressive therapy. Superimposed acute viral hepatitis can occur in endemic areas. PMID- 11422622 TI - Role of UGT1A1 mutation in fasting hyperbilirubinemia. AB - BACKGROUND AND AIM: Low-grade fasting hyperbilirubinemia is a common observation in healthy subjects (HS), whereas high-grade fasting hyperbilirubinemia is believed to be a characteristic finding of Gilbert's syndrome. This study was undertaken to assess the role of mutation in bilirubin UDP- glycosyltransferase gene (UGT1A1) on fasting hyperbilirubinemia. METHODS: Analysis of UGT1A1 and a caloric restriction test (400 kcal for 24 h) were performed in 56 healthy subjects (25 males, 31 females), and 28 patients with Gilbert's syndrome (18 males, 10 females). There were 29 healthy subjects with no mutation in UGT1A1, and 27 healthy subjects and 26 Gilbert's syndrome patients with mutations in the coding and/or promoter (TATA box) regions of UGT1A1. RESULTS: The mean increment of serum bilirubin (DeltaSB) was 7.6 micromol/L [corrected] (males) and 4.1 micromol/L (females) in subjects with no UGT1A1 mutation. Subjects with mutation in UGT1A1 showed higher levels of DeltaSB than individuals without mutation. Among healthy subjects, gender difference in DeltaSB values was observed only in individuals with the wild type of UGT1A1, but not in those with mutations in this gene. CONCLUSION: The results of the present study suggest that UGT1A1 mutation has a role in the development of high-grade fasting hyperbilirubinemia after caloric restriction. PMID- 11422623 TI - Effects and mechanisms of somatostatin analogs on apoptosis of pancreatic acinar cells in acute pancreatitis in mice. AB - AIM: To investigate the effects of somatostatin analogs (SSa) on apoptosis of pancreatic acinar cells and apoptosis-regulated gene bax, and p53 in treating acute pancreatitis in mice. METHODS: In cerulein-induced pancreatitis, with or without treatment of somatostatin, analogs (Octreotide) in CD-1 (BALB/c x DBetaAlpha/1) mice, apoptosis of pancreatic acinar cells was detected by using the TdT-mediated dUTP nick-end labeling (TUNEL) method, and the expression of apoptosis-regulated gene bax and p53 was determined by using the streptavidin peroxidase immunohistochemical technique and the RT-PCR method, respectively. RESULTS: On HE staining, acinar cells in the pancreas showed pyknotic nuclei and the formation of apoptotic bodies, which are the typical morphological features of apoptosis. Regarding TUNEL use, the apoptotic index of pancreatic acinar cells in the non-treated group at 5 and 14 h after induction of acute pancreatitis was significantly lower than those of the SSa-treated group, respectively (P < 0.01). On immunohistochemistry and RT-PCR, there was an expression of neither bax nor p53 in normal pancreatic tissues. The expression of bax in the SSa-treated group at 5 and 14 h after treatment of SSa was markedly higher than those of the non treated group, respectively (P < 0.01), but there was no significant difference in the expression of p53 between the SSa-treated group and the non-treated group. CONCLUSIONS: The induction of apoptosis in pancreatic acinar cells injury to reduce inflammatory reaction might be one of the mechanisms of SSa in treating acute pancreatitis in mice, and the mechanisms of apoptosis probably correlated with the expression of apoptosis-regulated gene bax, but have no relationship with the expression of p53. PMID- 11422624 TI - A meta-analysis of peritoneal lavage for acute pancreatitis. AB - BACKGROUND AND AIM: It has been postulated that continuous irrigation of the peritoneal cavity with crystalloid solutions in patients with acute pancreatitis can improve mortality and morbidity. The aim of the study is to perform a meta analysis of available randomized prospective clinical trials, to evaluate whether lavage influences mortality and morbidity in patients with acute pancreatitis. METHODS: We performed a computer search of Medline for all available literature on the use of lavage in patients with acute pancreatitis. A meta-analysis was conducted on eight randomized, prospective, clinical trials (a total of 333 patients) evaluating continuous peritoneal lavage in patients with acute pancreatitis. The end-points were mortality and morbidity (i.e. pancreatic necrosis, peripancreatic fluid collections, intra-abdominal abscess formation, septicemia, organ system failure). RESULTS: Continuous lavage did not improve either mortality (weighted mean difference 1.6%, 95% CI -6.7% to 9.9%, not significant (n.s.)) or morbidity (weighted mean difference 6.2%, 95% CI -3.2% to 15.6%, n.s.) when compared with control patients. CONCLUSIONS: The use of continuous peritoneal lavage in patients with acute pancreatitis has not been found to be associated with any significant improvement in mortality or morbidity. PMID- 11422625 TI - Hepatobiliary and pancreatic: a woman with abdominal pain. Giant hemangioma of the liver. PMID- 11422626 TI - Gastrointestinal: Small bowel enterolith ileus. PMID- 11422628 TI - Wilson's disease presenting with rapidly progressive visual loss: another neurologic manifestation of Wilson's disease? AB - Wilson's disease (WD) is a rare autosomal recessive disorder of copper metabolism resulting in copper-induced tissue damage that primarily involves the liver and central nervous system. The neurologic manifestations of WD almost universally involve a derangement of basal ganglia function or psychiatric disturbance. We report the case of a 46-year-old man presenting with end-stage liver disease caused by WD who had associated rapidly progressive optic neuropathy. We also discuss the possible association between the two conditions. PMID- 11422629 TI - Cardiac tamponade caused by spontaneous rupture of mediastinal lymph node metastasis of hepatocellular carcinoma. AB - Rupture of a hepatocellular carcinoma (HCC) is a well-known cause of death in patients with HCC. This report describes a rare case of HCC presenting as cardiac tamponade caused by a spontaneous rupture of mediastinal lymph node metastasis into the pericardial space. A transcatheter arterial embolization (TAE) of internal thoracic artery successfully controlled the bleeding, and the patient was rescued from cardiac tamponade. Although there was no rebleeding, the patient died from liver failure 2 months later. An autopsy revealed a poorly differentiated HCC in the liver, lung and mediastinal lymph nodes. PMID- 11422630 TI - Post-partum acute exacerbation of chronic hepatitis in a hepatitis C-carrier mother. AB - Hepatitis C virus infection is a global health problem; however, the interaction between pregnancy and chronic hepatitis C remains controversial. A Taiwanese woman with chronic hepatitis C had an uncomplicated pregnancy and gave birth to a female baby through spontaneous vaginal delivery. The serum levels of alanine aminotransferase and hepatitis C virus (HCV) RNA were measured before pregnancy, in the first and third trimesters, 1 and 3 months after delivery, respectively. During her pregnancy, the serum aminotransferase levels became normalized, while the serum HCV-RNA levels declined significantly and consecutively with the lowest viral load in the third trimester. One month after delivery, she had an abrupt elevation of serum HCV-RNA level, paralleling a hepatitis flare with serum aminotransferase level more than 20-fold the upper normal limit. The serum HCV RNA levels declined thereafter, and serum aminotransferase levels became normalized 3 months postdelivery. She was infected with HCV genotype 1a throughout the entire follow-up period, and other causes of hepatitis flare were excluded. In conclusion, post-partum acute exacerbation of chronic hepatitis may occur in HCV-carrier mothers, and an abrupt elevation of serum HCV-RNA level may be associated with the acute exacerbation. PMID- 11422631 TI - Influence of physician specialty on adoption and relinquishment of calcium channel blockers and other treatments for myocardial infarction. AB - OBJECTIVE: Recent reports have linked calcium channel blockers (CCBs) with an increased risk of acute myocardial infarction (AMI). We sought to determine to what extent physicians relinquished CCBs following these adverse reports and if there were differences in the use of CCBs and other AMI therapies across 3 levels of specialist involvement: generalist attendings, collaborative care (generalist with cardiologist consultation), and cardiologist attendings. DESIGN: We measured use of CCBs during hospitalization for AMI before (1992--1993) and after (1995- 1996) the adverse CCB reports, controlling for hospital-, physician-, and patient level variables. We also examined use of effective medications (aspirin, beta blockers, thrombolytic therapy) and ineffective AMI treatments (lidocaine). SETTING: Thirty-seven community-based hospitals in Minnesota. PATIENTS: Population-based sample of 5,347 patients admitted with AMI. MEASUREMENTS: The primary outcome was prescription of a CCB at the time of discharge from hospital. Secondary outcomes included use of other effective and ineffective AMI therapies during hospitalization and at discharge. MAIN RESULTS: Compared with cardiologists, generalist attendings were less likely to use aspirin (37% vs 68%; adjusted odds ratio [OR], 0.58; 95% confidence interval [95% CI], 0.42 to 0.80) and thrombolytics (29% vs 64%; adjusted OR, 0.18; 95% CI, 0.13 to 0.25), but not beta-blockers (20% vs 46%; adjusted OR, 0.93; 95% CI, 0.66 to 1.31). From 1992- 1993 to 1995--1996, the use of CCBs in patients with AMI decreased from 24% to 10%, the net result of physicians starting CCBs less often and discontinuing them more often. In multivariate models, the odds of CCB relinquishment after the adverse reports (adjusted OR, 0.33; 95% CI, 0.27 to 0.39) were independent of, and not modified by, the involvement of a cardiologist. CONCLUSIONS: Compared with cardiologists, generalist physicians were less likely to adopt some effective AMI therapies, particularly those associated with risk such as thrombolytic therapy. However, generalists were as likely as cardiologists to relinquish CCBs after the adverse reports. This pattern of practice may be the generalist physicians' response to an expanding, but increasingly risky and uncertain, pharmacopoeia. PMID- 11422632 TI - Relation of physician specialty and HIV/AIDS experience to choice of guideline recommended antiretroviral therapy. AB - BACKGROUND: Controversy exists regarding who should provide care for those with HIV/AIDS. While previous studies have found an association between physician HIV experience and patient outcomes, less is known about the relationship of physician specialty to HIV/AIDS outcomes or quality of care. OBJECTIVE: To examine the relationship between choice of appropriate antiretroviral therapy (ART) to physician specialty and HIV/AIDS experience. DESIGN: Self-administered physician survey. PARTICIPANTS: Random sample of 2,478 internal medicine (IM) and infectious disease (ID) physicians. MEASUREMENTS: Choice of guideline-recommended ART. RESULTS: Two patients with HIV disease, differing only by CD4+ count and HIV RNA load, were presented. Respondents were asked whether ART was indicated, and if so, what ART regimen they would choose. Respondents' ART choices were categorized as "recommended" or not by Department of Health and Human Services guidelines. Respondents' HIV/AIDS experience was categorized as moderate to high (MOD/HI) or none to low (NO/LO). For Case 1, 72.9% of responding physicians chose recommended ART. Recommended ART was more likely (P <.01) to be chosen by ID physicians (88.2%) than by IM physicians (57.1%). Physicians with MOD/HI experience were also more likely (P <.01) to choose recommended ART than those with NO/LO experience. Finally, choice of ART was examined using logistic regression: specialty and HIV experience were found to be independent predictors of choosing recommended ART (for ID physicians, odds ratio [OR], 4.66; 95% confidence interval [95% CI], 3.15 to 6.90; and for MOD/HI experience, OR, 2.05; 95% CI, 1.33 to 3.16). Results for Case 2 were similar. When the analysis was repeated excluding physicians who indicated they would refer the HIV "patient," specialty and HIV experience were not significant predictors of choosing recommended ART. CONCLUSIONS: Guideline-recommended ART appears to be less likely to be chosen by generalists and physicians with less HIV/AIDS experience, although many of these physicians report they would refer these patients in clinical practice. These results lend support to current recommendations for routine expert consultant input in the management of those with HIV/AIDS. PMID- 11422633 TI - Drug dosage in patients with renal failure optimized by immediate concurrent feedback. AB - OBJECTIVE: To examine the impact of immediate concurrent feedback on dose adjustment in patients with renal failure. DESIGN: Prospective 12-month study in patients with various degrees of renal failure, with comparison to a retrospective control group. SETTING: A 39-bed unit of a university hospital providing primary and tertiary care. PATIENTS: Patients with renal failure (estimated creatinine clearance < or = 50 mL/min) receiving at least 1 pharmacologically active drug. INTERVENTIONS: Education of physicians and immediate concurrent feedback on the ward giving estimated creatinine clearance and dose recommendations for renally eliminated drugs adjusted to individual renal function. MEASUREMENTS AND MAIN RESULTS: The percentage of dosage regimens adjusted to renal function and cost assessment of drug therapy were calculated. Overall, 17% of the patients had at least 1 estimated creatinine clearance < or = 50 mL/min. In the intervention group, the dose of 81% of renally eliminated drugs was adjusted to renal function, compared with 33% in the control group ( P <.001). The mean difference in cost between standard and adjusted dose of renally eliminated drugs in the intervention and control groups was 5.3 +/- 12.3 and 0.75 +/- 2.8 Swiss francs (approximately US$3.5 and US$0.5), respectively ( P <.001), accounting for 16.5% and 2.8%, respectively, of daily medication costs of all drugs. CONCLUSIONS: The proportion of doses of renally eliminated drugs adjusted to renal function can be substantially increased by immediate concurrent feedback. This saves drug costs and has the potential to prevent adverse drug reactions. PMID- 11422634 TI - Antibiotic use in Ontario facilities that provide chronic care. AB - OBJECTIVE: To determine the incidence and variability of antibiotic use in facilities which provide chronic care and to determine how often clinical criteria for infection are met when antibiotics are prescribed in these facilities. DESIGN: A prospective, 12-month, observational cohort study. SETTING: Twenty-two facilities which provide chronic care in southwestern Ontario. PARTICIPANTS: Patients who were treated with systemic antibiotics over the study period. MEASUREMENTS: Characteristics of antibiotic prescriptions (name, dose, duration, and indication) and clinical features of randomly selected patients who were treated with antibiotics. RESULTS: A total of 9,373 courses of antibiotics were prescribed for 2,408 patients (66% of all patients in study facilities). The incidence of antibiotic prescriptions in the facilities ranged from 2.9 to 13.9 antibiotic courses per 1,000 patient-days. Thirty-six percent of antibiotics were prescribed for respiratory tract infections, 33% for urinary infections, and 13% for skin and soft tissue infections. Standardized surveillance definitions of infection were met in 49% of the 1,602 randomly selected patients who were prescribed antibiotics. Diagnostic criteria for respiratory, urinary, and skin infection were met in 58%, 28%, and 65% of prescriptions, respectively. One third of antibiotic prescriptions for a urinary indication were for asymptomatic bacteriuria. Adverse reactions were noted in 6% of prescriptions for respiratory and urinary infections and 4% of prescriptions for skin infection. CONCLUSIONS: Antibiotic use is frequent and highly variable amongst patients who receive chronic care. Reducing antibiotic prescriptions for asymptomatic bacteriuria represents an important way to optimize antibiotic use in this population. PMID- 11422635 TI - Prevalence and significance of unrecognized lower extremity peripheral arterial disease in general medicine practice*. AB - OBJECTIVE: To determine the prevalence of unrecognized lower extremity peripheral arterial disease (PAD) among men and women aged 55 years and older in a general internal medicine (GIM) practice and to identify characteristics and functional performance associated with unrecognized PAD. DESIGN: Cross-sectional. SETTING: Academic medical center. PARTICIPANTS: We identified 143 patients with known PAD from the noninvasive vascular laboratory, and 239 men and women aged 55 and older with no prior PAD history from a GIM practice. Group 1 consisted of patients with PAD consecutively identified from the noninvasive vascular laboratory (n = 143). Group 2 included GIM practice patients found to have an ankle brachial index less than 0.90, consistent with PAD (n = 34). Group 3 consisted of GIM practice patients without PAD (n = 205). MEASUREMENTS AND MAIN RESULTS: Leg functioning was assessed with the 6-minute walk, 4-meter walking velocity, and Walking Impairment Questionnaire (WIQ). Of GIM practice patients, 14% had unrecognized PAD. Only 44% of patients in Group 2 had exertional leg symptoms. Distances achieved in the 6-minute walk were 1,130, 1,362, and 1,539 feet for Groups 1, 2, and 3, respectively, adjusting for age, gender, and race (P <.001). The degree of difficulty walking due to leg symptoms as reported on the WIQ was comparable between Groups 2 and 3 and significantly greater in Group 1 than Group 2. In multiple logistic regression analysis including Groups 2 and 3, current cigarette smoking was associated independently with unrecognized PAD (odds ratio [OR], 6.82; 95% confidence interval [95% CI], 1.55 to 29.93). Aspirin therapy was nearly independently associated with absence of PAD (OR, 0.37; 95% CI, 0.12 to 1.12). CONCLUSION: Unrecognized PAD is common among men and women aged 55 years and older in GIM practice and is associated with impaired lower extremity functioning. Ankle brachial index screening may be necessary to diagnose unrecognized PAD in a GIM practice. PMID- 11422636 TI - The evaluation of two methods to facilitate shared decision making for men considering the prostate-specific antigen test. AB - OBJECTIVE: California law (Grant H. Kenyon Prostate Cancer Detection Act) requires physicians to inform all patients older than aged 50 years who receive a prostate examination about the availability of the prostate-specific antigen (PSA) test. Physicians are not given guidance on how this information should be presented. We sought to evaluate the effects upon PSA screening rates of informing patients about PSA testing by 2 different techniques. DESIGN: Factorial comparison of discussion versus video formats for presenting information about the PSA test. SETTING: Patients were recruited through the Health Appraisal screening program in the Department for Preventive Medicine, Kaiser Permanente, San Diego, Calif. PARTICIPANTS: Male patients undergoing health appraisal screening participated in 1 of 4 groups providing information about PSA screening: usual care ( n=43), discussion about risks and benefits of PSA ( n=45), shared decision-making video ( n=46), or video plus discussion ( n=42). Participants were sequentially assigned to 1 of the 4 groups. RESULTS: No significant differences in demographics or family history was demonstrated between the groups at the time of group assignment. Participants in the intervention groups rated the information as clear, balanced, and fair. There were significant differences in the number of men requesting a PSA test, with the highest rate in the usual care group (97.7%), followed by discussion (82.2%), video (60.0%), and video plus discussion (50.0%). CONCLUSION: Providing information about PSA screening in the form of video or discussion is feasible and significantly alters PSA screening rates. PMID- 11422637 TI - Teaching resource and information management using an innovative case-based conference. AB - Physicians play a critical role in controlling resource use in medicine. This paper describes an innovative, interdisciplinary conference that teaches housestaff and medical students about resource and information management in the hospital setting. The objectives are to help foster communication between physicians and other members of the health care team, to improve the understanding of hospital reimbursement, and to influence attitudes toward practicing cost effectiveness. The conference structure includes the following components: case presentation by the treating physician and follow-up information provided by the primary care physician, a review of the itemized hospital bill, discussion of coding issues, discussion of hospital reimbursement comparing case data to institutional and state averages, and a summary of key take-home points and lessons. PMID- 11422638 TI - Is food insufficiency associated with health status and health care utilization among adults with diabetes? AB - OBJECTIVES: Preliminary studies have shown that among adults with diabetes, food insufficiency has adverse health consequences, including hypoglycemic episodes and increased need for health care services. The purpose of this study was to determine the prevalence of food insufficiency and to describe the association of food insufficiency with health status and health care utilization in a national sample of adults with diabetes. METHODS: We analyzed data from adults with diabetes ( n =1,503) interviewed in the Third National Health and Nutrition Examination Survey. Bivariate and multivariate analyses were used to examine the relationship of food insufficiency to self-reported health status and health care utilization. RESULTS: Six percent of adults with diabetes reported food insufficiency, representing more than 568,600 persons nationally (95% confidence interval, 368,400 to 768,800). Food insufficiency was more common among those with incomes below the federal poverty level (17% vs 4%, P < or = .001). Adults with diabetes who were food insufficient were more likely to report fair or poor health status than those who were not (63% vs 43%; odds ratio, 2.2; P=.05). In a multivariate analysis, fair or poor health status was independently associated with poverty, nonwhite race, low educational achievement, and number of chronic diseases, but not with food insufficiency. Diabetic adults who were food insufficient reported more physician encounters, either in clinic or by phone, than those who were food secure (12 vs 7, P<.05). In a multivariate linear regression, food insufficiency remained independently associated with increased physician utilization among adults with diabetes. There was no association between food insufficiency and hospitalization in bivariate analysis. CONCLUSIONS: Food insufficiency is relatively common among low-income adults with diabetes and was associated with higher physician utilization. PMID- 11422639 TI - "I can't afford that!": dilemmas in the care of the uninsured and underinsured. AB - When patients lack sufficient health care insurance, financial matters become integrally intertwined with biomedical considerations in the process of clinical decision making. With a growing medically indigent population, clinicians may be compelled to bend billing or reimbursement rules, lower standards, or turn patients away when they cannot afford the costs of care. This article focuses on 3 types of dilemmas that clinicians face when patients cannot pay for needed medical services: (1) whether to refer the individual to a safety net provider, such as a public clinic; (2) whether to forgo indicated tests and therapies because of cost; and (3) whether to reduce fees by fee waivers or other adjustments in billing. Clinicians' responses to these dilemmas impact on quality of care, continuity, safety net providers, and the liability risk of committing billing violations or offering nonstandard care. Caring for the underinsured in the current health care climate requires an understanding of billing regulations, a commitment to informed consent, and a beneficent approach to finding individualized solutions to each patient care/financial dilemma. To effect change, however, physicians must address issues of social justice outside of the office through political and social activism. PMID- 11422640 TI - Teaching about cost-effective use of medical resources: still trying after all these years. PMID- 11422641 TI - Differences between generalists and specialists: knowledge, realism, or primum non nocere? PMID- 11422642 TI - The contribution of science in facilitating the inclusion of people with intellectual disability into the community. PMID- 11422643 TI - Sexual abuse in children and adolescents with intellectual disability. AB - The present authors conducted a study of the occurrence of victimization and the perpetration of sexual abuse among 43 in-patients with intellectual disability aged between 9 and 21 years who were admitted to a child and adolescent psychiatric in-patient department over a period of 5 years. A retrospective case note review was employed that explored the nature and severity of abuse in relation to the age, gender and level of disability. The prevalence of abuse or abusive behaviour, i.e. 14% of 300 admissions, did not change over time. In 13 out of the 43 cases, the issue of sexual abuse was identified after admission. Victimization alone occurred in 21 cases, perpetration alone in six cases, and both victimization and perpetration in 16 cases. Fifty per cent of the victims had been abused by a member of their close or extended family. Most cases (62%) were adolescents. There was only one instance of a victim being abused by a female. However, there were five girls who were perpetrators, all of whom had previously been victims. By contrast, 11 out of the 17 male perpetrators had been victims. Despite difficulties of disclosure, it was possible to establish that severely disabled patients had suffered sexual abuse. The present data support theories which (1) recognize gender differences in sexual abuse patterns and (2) have a developmental perspective, incorporating the influence of adolescence. PMID- 11422644 TI - Self-assessment of relationships with peers in children with intellectual disability. AB - Interaction with peers is important for the development of children, but children with special needs may feel rejected by their peers. The present study examines self-assessment of relationships with peers by children with intellectual disability (ID; n = 20) and children from the general population (n = 20). All participants attended a regular primary school and were aged between 7 and 10.5 years; both sexes were represented in the samples. The Behavior Rating Profile was applied. The results of the children with ID on the "Student Rating Scale: Peers" did not show statistically significant differences from the results of children from general population, with both groups responding similarly to the self-perception scale. However, the sociometric results obtained from their peers clearly show that children with ID are not accepted by their classmates. Peers frequently and more often refuse to study, sit together in class or socialize after classes with children with ID than is the case for children without ID. Despite the high frequency of rejection, it is concluded that children with ID of younger primary school age have average confidence in their own abilities and in the success of their relationships with their peers. Further education among the children who reject them could have a negative impact on their self-esteem. PMID- 11422645 TI - The prevalence of constipation in institutionalized people with intellectual disability. AB - Constipation is a common problem in people with intellectual disability (ID). Laxatives are frequently prescribed with disappointing results. The prevalence of constipation was investigated in a random population of 215 people with ID (IQ < 50) and constipation was correlated with clinical symptoms. All subjects were scored for bowel habits. Constipation was defined as having a bowel movement less than three times a week or the necessity of using laxatives more than three times a week. Further possible accompanying factors were evaluated. The control subjects were defined as individuals who did not use laxatives. Subjects with constipation were defined as patients and were compared to subjects without constipation (controls). One hundred and forty-nine out of 215 cases (69.3%) showed constipation. Constipation was significantly correlated with non ambulancy, cerebral palsy, the use of anticonvulsive medication or benzodiazepines, H2-receptor antagonists or proton pump inhibitors, food refusal, and an IQ < 35. Fifty-eight per cent of the patients used bisacodyl or magnesium oxide, 39% lactulose, 13% sodiumlaurylsulphoacetate/sodium citrate/sorbitol and only 10% were given sodium phosphate enemas. Faecal soiling was found in 15% of subjects, while manual evacuation of faeces was performed in nearly 7% of cases. Constipation was randomly demonstrated in almost 70% of the population with ID. Subjects with the above-mentioned accompanying factors are especially at risk for constipation. Contrary to the general population, constipation in people with ID is associated with little use of phosphate enemas, microlax, a low incidence of faecal soiling and manual evacuation of faeces, suggesting an aetiology without distal faecal impaction. The regimen and effect of therapy has to be studied to define adequate treatment schedules. PMID- 11422646 TI - Helicobacter pylori: has the killer escaped from the institution? A possible cause of increased stomach cancer in a population with intellectual disability. AB - Advancing knowledge of the existence of Helicobacter pylori and its association with gastrointestinal tract malignancy, and previous research showing higher-than expected gastrointestinal tract malignancy in institutionalized adults with intellectual disability (ID) prompted a review of all deaths as a result of cancer in the Stoke Park group of hospitals for people with ID between 1946 and 1996. A 50-year, retrospective case note analysis of all deaths from cancer in an institution for people with ID was undertaken. Death from stomach cancer accounted for up to 48% of all cancer deaths. A further 25 residents had died of perforated stomach ulcers. The higher proportion of deaths specifically caused by stomach cancer in a population with ID has not been noted previously. It is postulated that the high levels of H. pylori infection found in institutionalized populations may be instrumental in this higher mortality rate and that the closure of the institutions without evaluation of H. pylori status transfers the problem unresolved to the community. Existing guidelines for the screening and eradication of H. pylori developed for the general population are inadequate when applied to people with ID, and therefore, the value of population screening and mass eradication programmes is explored. PMID- 11422647 TI - Audit and comprehensive health assessment programme in the primary healthcare of adults with intellectual disability: a pilot study. AB - International research has demonstrated significant shortcomings in the health of adults with intellectual disability (ID). Because general practitioners (GPs) are the main providers of primary healthcare for this population, strategies to improve general practice care are an important aspect of rectifying these shortcomings. The present pilot study aimed to determine the effect of various interventions on health maintenance activities and to assess their acceptability to GPs, with a view to informing larger scale studies. The GPs were recruited through an earlier questionnaire-based postal survey. The GPs identified all their adult patients with ID, then obtained consent for participation from three patients randomly selected by the investigators. The GPs completed two self evaluation forms and case note audits 12 months apart, read a synopsis of the relevant literature provided by the researchers, and completed a comprehensive health assessment (CHA) of their three patients. Forty-five GPs agreed to participate in the CHA programme (CHAP), and 15 completed the project. Thirty eight patients completed the project. The number of patient-GP dyads who completed the project was too small to demonstrate statistically significant changes in health issues over time. The GPs found that the synopsis of the literature was the best intervention for increasing knowledge and was also the most practical to use in general practice. The CHAP was the intervention that prompted the most action from the GP which would not have been undertaken otherwise. The CHAP appeared to provide a superior review process compared to the other interventions used in the present study. The numbers of health maintenance activities found to be overdue and the number of health issues detected as a result of the process were considerable. The CHAP served as a communication tool and an educative instrument, providing a basis for future studies and strategies to improve the general practice care of adults with ID. PMID- 11422648 TI - Complexity of staff communication and reported level of understanding skills in adults with intellectual disability. AB - Staff reports of the communication acts taking place with 22 adults with intellectual disability were compared with video observations of the communication acts used by staff with 12 of these service users. The interactions were coded in terms of the form of communication used, the function of the act and the level of complexity. The results show that staff tend to underestimate their own use of verbal communication and overestimate their use of non-verbal communication. The findings also indicate a mismatch between the reported level of understanding of the service user and the level of complexity of the language used. Staff appeared unable to adapt their communication to the skills of the service user and an average of 45% of communicative acts were outside the reported understanding skills of the individual. The implications of these findings are discussed and possible explanations for staff behaviour are suggested. PMID- 11422649 TI - Psychopathology of sexual abuse in young people with intellectual disability. AB - The present study addressed two issues using a sample of child and adolescent victims and perpetrators of sexual abuse: (1) the extent of post-traumatic stress disorder (PTSD) in the sample; and (2) the possible distinction between perpetrators whose motivations were sexually impulsive, and those who were controlling and abuse-reactive. Retrospective case material from 43 cases (21 victims only, and 22 perpetrators, of whom 16 were also victims) provided the data. Post-traumatic symptomatology was not common. Only one case of PTSD was found. Perpetrators could be distinguished by whether they had suffered sexual abuse alone, sexual and physical abuse, or neither. The proposed distinction between perpetrators received support. Sexual abuse directed at younger victims was associated with earlier experience of multiple forms of abuse. The present data does not support the view that post-traumatic symptoms following victimization are a mediator of sexual abuse perpetration. It is argued that an elaboration of the Williams & New developmental model of perpetration better fits the data. PMID- 11422650 TI - Breast cancer screening for older women with intellectual disability living in community group homes. AB - Breast cancer is one of the commonest cancers to affect women. Present health service guidelines call for screening and mammography for all women aged between 50 and 65 years in an effort to increase early detection and improve survival rates. Nulliparity is one of the associated risk factors for breast cancer. Women with intellectual disability (ID) are increasing in longevity and are frequently nulliparous, and therefore, they are at increased risk of developing breast cancer. The aim of the present study was to review the uptake and knowledge of women with ID living in the community of breast screening programmes. A postal survey of women aged > or = 50 years with ID living in community group homes was used to gather data. Only one-third of the women carried out regular breast examination and a similar proportion had received invitations to mammography. General practitioners and practice nurses were currently playing very minor roles in breast screening these women. Primary health care professionals may be missing opportunistic health promotion opportunities and the support services for women with ID living in the community could be provided with better training and resources to improve breast cancer screening in this vulnerable group. PMID- 11422651 TI - Factors associated with intended staff turnover and job search behaviour in services for people with intellectual disability. AB - Staff turnover is a major problem in services for people with intellectual disability (ID). Therefore, understanding the reasons for staff turnover is vital for organizations seeking to improve their performance. The present study investigates the factors directly and indirectly associated with an intention to leave an organization and actual job search behaviour amongst staff in services for people with ID. As part of a large-scale survey of staff in services for people with ID, information was collected from 450 staff concerning intended turnover, job search behaviour and a wide range of factors potentially associated with these outcomes. Path analyses revealed that work satisfaction, job strain, younger staff age and easier subjective labour conditions were directly associated with intended turnover. The same factors, with the exception of younger staff age, were also directly associated with job search behaviour. Factors indirectly associated with these outcomes included wishful thinking, alienative commitment to the organization, lack of staff support, role ambiguity, working longer contracted hours, having a low-status job, a lack of influence over decisions at work and less orientation to working in community settings with people with ID. The models of staff turnover empirically derived in the present study confirm and extend previous research in this area. The implications for organizations are discussed. PMID- 11422652 TI - A microswitch for vocalization responses to foster environmental control in children with multiple disabilities. AB - The present study assessed whether two children with multiple disabilities could learn to successfully operate a special microswitch through vocalization responses so as to obtain environmental stimulation. The study was carried out using a non-concurrent multiple baseline design across individuals. The treatment phase of the second child started after he had received twice the number of baseline sessions used for the first child. The results were positive, with both children increasing the frequencies of their vocalization responses during the treatment. The importance of using such responses (which do not require excessive efforts and are highly valued) for operating a microswitch with children with multiple disabilities is discussed. PMID- 11422654 TI - The role of gene-environment interaction in the aetiology of human cancer: examples from cancers of the large bowel, lung and breast. AB - It has become increasingly clear that cancer can be considered neither purely genetic nor purely environmental. A relatively new area of cancer research has focused on the interaction between genes and environment in the same causal mechanism. Primary candidates for gene-environment interaction studies have been genes that encode enzymes involved in the metabolism of established cancer risk factors. There are common variant forms of these genes (polymorphisms), which may alter metabolism and increase or decrease exposure to carcinogens, thus impacting the risk of cancer. We present an overview of enzymes involved in carcinogen metabolism, present epidemiological tools to evaluate gene-environment interactions, and provide examples from cancers of the breast, lung and large bowel. PMID- 11422655 TI - The relationship between educational level and mortality. The Reykjavik Study. AB - OBJECTIVE: Epidemiological studies have indicated an association between socioeconomic factors and health. It has not been clearly established whether this association is wholly or partly independent of classical risk factors. Our objective was to estimate the relationship between educational level and coronary artery disease (CAD), mortality and all-cause mortality. The Reykjavik Study involving 18 912 participants followed-up 4-30 years provides an ideal opportunity to address this question. DESIGN AND SUBJECTS: The participants were aged 33-81 years and living in the Reykjavik area. They were divided into four groups according to education. The standard risk factors were assessed on entry and mortality, and cause of death registered during follow-up. Multiple Cox regression analysis was applied to assess the relationship between age at examination, year of examination, educational level and mortality. RESULTS: The all-cause mortality and CAD mortality was significantly related to education, even after adjustment for classical risk factors. For men, 14% (95% CI: 2-24) reduction was found in CAD mortality for those having high school education relative to elementary school. The figures for junior college and university education were 17% (95% CI: 1-31) and 38% (95% CI: 21-32), respectively. These figures were only slightly lower when major CAD risk factors were controlled for and still significant. Similar figures were found for all-cause mortality. For women 34% (95% CI: 18-48) reduction was found in CAD mortality for high school education and 55% (95% CI: 22-74) for junior college, but too few had university education for reliable results. The figures were lower for all-cause mortality, but significant. The figures were reduced when major CAD risk factors were controlled for, but still significant. CONCLUSION: Education is a strong protective factor both for all-cause and CAD mortality. Only a small part of this effect can be explained through conventional risk factors. PMID- 11422656 TI - Increased levels of tPA antigen and tPA/PAI-1 complex in myotonic dystrophy. AB - OBJECTIVE: To assess the fibrinolytic system in myotonic dystrophy (DM1), a disease connected to features of the metabolic syndrome, including a prominent insulin resistance, increased body fat mass, and hypertriglyceridaemia. We hypothesized that abnormalities in the fibrinolytic system are linked to metabolic dysfunction in DM1. DESIGN: Circulating morning levels of tissue plasminogen activator (tPA) and plasminogen activator inhibitor type 1 (PAI-1) antigens, tPA/PAI-1 complex, lipids and insulin were determined. Genetic analyses, including calculation of allele size, were performed in all patients. Body fat mass was estimated with bioelectrical impedance analysis. SETTING: Out patient clinic in collaboration with Umea University Hospital. SUBJECTS: A total of 42 otherwise healthy patients with DM1 (22 men, 20 women; median age 41.5 years) and 50 controls (27 men, 23 women; median age 42.0 years). MAIN OUTCOME MEASURES: The tPA and PAI-1 antigens, tPA/PAI-1 complex, blood lipids and body fat mass. RESULTS: The tPA antigen and tPA/PAI-1 complex levels were significantly increased in DM1 patients (P < 0.001 and P < 0.05, respectively) whilst levels of PAI-1 did not differ from controls. Triglyceride levels were increased (P < 0.001) whereas HDL cholesterol levels were lower in DM1 patients (P < 0.05). Body fat mass was increased in DM1 patients (P < 0.001). CONCLUSIONS: The fibrinolytic system is disturbed in DM1 patients, with increased levels of tPA and tPA/PAI-1 complex but paradoxically unaltered levels of PAI-1, in spite of a severely increased body fat mass. This may imply an abnormal function of adipose tissue in DM1, and calls for further studies of the fibrinolytic system in this disease.bstra PMID- 11422657 TI - Plasma fibrinogen level and long-term prognosis in Norwegian middle-aged patients with previous myocardial infarction. A 10 year follow-up study. AB - OBJECTIVES: To investigate the prognostic value of plasma fibrinogen level amongst middle-aged survivors of myocardial infarction (MI). DESIGN: Prospective cohort study. SETTINGS: Determination of fibrinogen and other prognostic variables in MI patients recruited in a presumably stable phase of coronary heart disease (CHD). SUBJECTS: A total of 247 middle-aged CHD patients (54 women and 193 men) who had their first MI at age < or = 60 (women) or < or = 55 (men) were recruited at least 3 months after (mean 2.1 years) the most recent MI. MAIN OUTCOME MEASURES: The primary endpoint was total mortality, and the secondary endpoint was cardiac deaths. The tertiary endpoint was major cardiac events (cardiac death, MI and cardiac arrest). RESULTS: During a follow-up period of 10 years a total of 44 patients had died, 36 from cardiac causes. Major cardiac event occurred in 70 patients. After adjusting for age, ejection fraction (EF), total serum cholesterol (TC), smoking and hypertension, patients in the top quartile of fibrinogen (> or = 4.0 g L-1) had a relative risk (RR) of 1.8 (95% CI 1.0-3.6) (P = 0.07) for death of all causes. The top quartile of fibrinogen was a stronger predictor of cardiac death; RR = 2.2 (95% CI 1.1-4.4) (P = 0.03), whilst the effect on the endpoint major cardiac event was not significant; RR=1.1 (95% CI 0.6-1.9) (P = 0.69). CONCLUSIONS: A plasma fibrinogen level in the top quartile predicted cardiac death in middle-aged patients who had suffered MI. PMID- 11422658 TI - Microalbuminuria is common, also in a nondiabetic, nonhypertensive population, and an independent indicator of cardiovascular risk factors and cardiovascular morbidity. AB - OBJECTIVES: To assess the prevalence of microalbuminuria in the general population, especially in nondiabetic and nonhypertensive subjects, and its association with known cardiovascular risk factors and cardiovascular morbidity. DESIGN: Cross-sectional cohort study. SETTING: Inhabitants of the city of Groningen, the Netherlands. SUBJECTS: All inhabitants, aged between 28 and 75 years, were send a postal questionnaire and a vial to collect an early morning urine sample (n = 85 421). Of these 40 856 subjects (47.8%) responded. Cardiovascular risk factors and morbidity were validated in a well defined nondiabetic and nonhypertensive group of 5241 subjects. MAIN OUTCOME MEASURES: Microalbuminuria, self-reported cardiovascular risk and cardiovascular morbidity in the total study cohort, and additionally more detailed measurements in a subset of the total population. RESULTS: Microalbuminuria (20-200 mg L-1) was present in 7.2% of the subjects and independently associated with age, gender, hypertension, diabetes, smoking, previous myocardial infarction and stroke. Some of these associations were already observed at albuminuria levels of 10-20 mg L 1. After exclusion of the diabetic and hypertensive subjects, microalbuminuria was still prevalent in 6.6% of the subjects. CONCLUSIONS: Microalbuminuria appears to be common not only in the general population but also in a nondiabetic, nonhypertensive population and is independently associated with increased cardiovascular risk factors and cardio-vascular morbidity. Importantly, some of these associations are present at urinary albumin levels currently considered to be normal. These findings suggest that urinary albumin measurements may be useful in early risk profiling and prevention of cardiovascular disease in the population at large. PMID- 11422659 TI - Risk assessment in patients with acute myocardial infarction treated with thrombolytic therapy. AB - OBJECTIVE: Several noninvasive methods have prognostic information regarding mortality and new coronary events after an acute myocardial infarction (AMI). The practical for clinical decision-making in the immediate postmyocardial infarction (MI) period is, however, less evident. We investigated consecutive patients with AMI treated with thrombolysis to further clarify this issue. DESIGN: A total of 100 patients (27% women) aged 64 +/- 9 years (mean +/- SD) were studied. Risk assessment based on a clinical score system, myocardial perfusion scintigraphy single photon emission computed tomography (SPECT) at rest and during adenosine stress, echocardiography, radionuclide angiography, symptom-limited exercise stress test, and 24-h Holter ECG recording with ST-analysis and analysis of heart rate variability (HRV) were performed 5-8 days after hospital admission. Mortality, nonfatal reinfarction, and the need for revascularization were followed during 12 months. SETTING: A university hospital. RESULTS: A total of 6 patients died, seven had a nonfatal reinfarction, and 23 were revascularized. Inability to perform an exercise test (P = 0.004) and an ejection fraction (EF) < 40% (P = 0.002) were the only parameters separating those who died from the survivors. No method could predict a nonfatal reinfarction. Patients suffering either death or nonfatal reinfarction had a clinical risk assessment score 2 points higher (8.8 vs. 6.7, P = 0.05) than the group without such events. A positive symptom-limited exercise stress test (P = 0.027), ST-depressions on Holter ECG (P = 0.04), and reversibility on myocardial perfusion scintigraphy (P = 0.029) predicted the need for revascularization. CONCLUSION: Risk assessment based on clinical information, exercise stress testing, and an estimate of left ventricular function (e.g. via echocardiography) contribute with prognostic information in thrombolysed MI-patients. Additional noninvasive investigations such as adenosine-SPECT, analysis of HRV, and Holter-monitoring do not add to these commonly available tools in risk stratification of subjects at low to medium risk. PMID- 11422660 TI - Nicotine infusion acutely impairs insulin sensitivity in type 2 diabetic patients but not in healthy subjects. AB - OBJECTIVES: The aim of this study was to examine if an acute nicotine infusion alters insulin sensitivity to a similar degree in type 2 diabetic patients as in healthy control subjects. DESIGN: . Double-blind, cross-over, placebo-controlled, randomized experimental study. Nicotine 0.3 microg kg-1 min(-1) or NaCl was infused (2 h) during a euglycaemic hyperinsulinaemic clamp (4 h) to assess insulin sensitivity. SETTING: University research laboratory. SUBJECTS: Six male and female type 2 diabetic patients [DM2; age 54 +/- 10 (mean +/- SD) years; body mass index (BMI) 25.6 +/- 2.9 kg m(-2)] treated with diet or one oral hypoglycaemic agent and six age- and BMI-matched control subjects (Ctr). MAIN OUTCOME MEASURE: Insulin sensitivity (rate of glucose infusion per kg fat free body mass and minute), nicotine and free fatty acid (FFA) levels, pulse rate and blood pressure. RESULTS: The infusions produced similar nicotine levels in both groups. In the absence of nicotine, DM2 were more insulin resistant than Ctr (6.7 +/- 0.4 vs. 10.9 +/- 0.3 mg kg-1 LBM min(-1), respectively; P < 0.0001). This insulin resistance was further aggravated by the nicotine infusion in DM2 but not in Ctr (4.6 +/- 0.3 vs. 10.9 +/- 0.3 mg kg(-1) LBM min(-1); P < 0.0001). Only minor differences were seen in FFA levels, pulse rates and blood pressure. CONCLUSIONS: At this low infusion rate, nicotine aggravated the insulin resistance in DM2 but not in Ctr. This finding may be because of the (dysmetabolic) diabetic state per se or to an increased sensitivity to environmental factors associated with a genetic predisposition for type 2 diabetes. These results show that diabetic subjects are particularly susceptible to the detrimental effects of nicotine. PMID- 11422661 TI - Heart rate variability and n-3 polyunsaturated fatty acids in patients with diabetes mellitus. AB - OBJECTIVES: Dietary n-3 polyunsaturated fatty acids (PUFA) derived from fish may reduce the incidence of sudden cardiac death (SCD). The aim of the present study was to examine associations between n-3 PUFA and 24-h heart rate variability in patients with type 1 and type 2 diabetes mellitus (DM). DESIGN: Observational study. SETTING: The out-patient's diabetic clinic at Hjorring Hospital, Hjorring, Denmark. SUBJECTS: Forty-three patients with type 1 DM and 38 patients with type 2 DM. MAIN OUTCOME MEASURES: The patients fulfilled a food-questionnaire regarding fish consumption, whilst the content of n-3 PUFA in platelets was measured and 24-h heart rate variability (HRV) was obtained. RESULTS: The patients fish consumption was strongly related to their content of n-3 PUFA in platelets. Furthermore, in patients with type 1 DM a close positive association was found between the content of n-3 PUFA in platelets and 24-h HRV. This association was not significant in patients with type 2 DM. CONCLUSIONS: The positive association between n-3 PUFA in platelets and HRV may indicate a beneficial effect of n-3 PUFA on HRV in patients with type 1 DM. Further studies are warranted to clarify whether supplementation with n-3 PUFA reduce the risk of SCD amongst patients with DM. PMID- 11422662 TI - Large differences in serum leptin levels between nonwesternized and westernized populations: the Kitava study. AB - OBJECTIVES: To compare serum leptin between nonwesternized and westernized populations. SETTING: (i) The tropical island of Kitava, Trobriand Islands, Papua New Guinea and (ii) the Northern Sweden MONICA study population. Design. Cross sectional survey. METHODS: Fasting levels of serum leptin were analysed in 163 randomly selected Kitavans aged 20-86 years and in 224 Swedes aged 25-74. MAIN OUTCOME MEASURE: Mean and determinants of serum leptin. RESULTS: Geometric mean of serum leptin in Kitavan males and females were 1.5 and 4.0 vs. 4.9 and 13.8 ng mL-1 in Swedish male and females (P < 0.0001 for both sexes). In Kitavans, observed geometric mean were close to predicted levels (1.8 ng mL(-1) for males and 4.5 ng mL-1 for females) based on multiple linear regression equations including body mass index (BMI), triceps skinfolds (TSF) and age from the Swedish population-based sample. In Kitavans serum leptin was positively related to TSF amongst both sexes and, amongst females, to BMI. In Kitavans leptin was not related to fasting serum insulin. TSF explained 55% of the variation of leptin amongst females. There was a slight age-related increase of leptin amongst males. In Kitava leptin was not related to fasting serum insulin which was substantially lower than in Sweden. CONCLUSION: The low concentrations of serum leptin amongst Kitavans probably relates to the absence of overweight and hyperinsulinaemia. At a population level serum leptin can apparently be predicted from simple measures of adiposity. PMID- 11422663 TI - A high prevalence of hypovitaminosis D in Finnish medical in- and outpatients. AB - OBJECTIVE: To study the prevalence of hypovitaminosis D [serum 25(OH)D < or = 37 nmol L-1)] in Finnish medical in- and outpatients in a cross-sectional study. METHODS: The subjects were 106 consecutive medical inpatients (57 females, 49 males with mean ages of 65 and 58 years) from the Peijas Hospital, Vantaa, Finland, and 99 ambulatory patients (48 females, 51 males with mean ages of 42 and 46 years) contacting a private outpatient centre in Helsinki, Finland. Serum 25(OH)D, vitamin D binding protein (DBP), free vitamin D index (FDI), intact PTH (iPTH), and albumin-corrected calcium were measured. RESULTS: Serum 25 hydroxyvitamin D [25(OH)D] was 37 nmol L(-1) or less in 70% of female and in 61% of male inpatients and in 44% of female and in 37% of male outpatients. In the whole population, a statistically significant inverse association (P < 0.0001) was detected between iPTH and 25(OH)D levels; the iPTH concentration appeared to start increasing when 25(OH)D concentration was 50 nmol L(-1) or less. The association remained the same (P < 0.0001) when FDI was used instead of 25(OH)D in the calculations. When the sexes were analysed separately, the statistically significant association was found only in females (P < 0.0001 for iPTH versus 25(OH)D; P < 0.0001 for iPTH versus FDI) but not in males. CONCLUSION: Hypovitaminosis D is very common amongst Finnish in- and outpatients in both sexes, causing secondary hyperparathyroidism in females. More extensive studies are warranted to elucidate the vitamin D status of the Finnish population. PMID- 11422664 TI - Amiodarone-induced adult respiratory distress syndrome after nonthoracotomy subcutaneous defibrillator implantation. PMID- 11422665 TI - Improved spatial discrimination of protein reaction states in cells by global analysis and deconvolution of fluorescence lifetime imaging microscopy data. AB - The deconvolution of fluorescence lifetime imaging microscopy (FLIM) data that were processed with global analysis techniques is described. Global analysis of FLIM data enables the determination of relative numbers of molecules in different protein reaction states on a pixel-by-pixel basis in cells. The three-dimensional fluorescence distributions of each protein state can then be calculated and deconvolved. High-resolution maps of the relative concentrations of each state are then obtained from the deconvolved images. We applied these techniques to quantitatively image the phosphorylation state of ErbB1 receptors tagged with green fluorescent protein in MCF7 cells. PMID- 11422666 TI - Space-multiplexed multifocal nonlinear microscopy. AB - Standard forms of nonlinear microscopy rely on single beam scanning, but the usually weaker signal and the need to image in real-time call for parallelization of the image formation. Since the nonlinear susceptibilities necessitate a comparatively large illumination power, with current laser systems the brightness or field of view of any parallelized nonlinear microscope is limited by the brightness of the laser. For example, by producing an array of high aperture foci, multifocal multiphoton microscopy (MMM) provides real-time, light-efficient three-dimensional fluorescence imaging at high-resolution. The available power limits the degree of parallelization and hence codetermines the field of view. As the utilization of all the laser power is imperative, the focal intensity can be adjusted only through altering the number of foci. This compromises to some extent the flexibility to adjust the focal intensity to benign and effective levels. Here we introduce space-multiplexing (SMX) as a novel option in parallelized nonlinear microscopy, which enables an improved exploitation of the total laser power and facilitates changing the intensity levels in selected regions, without attenuating the total laser power. The basic idea of SMX is to overlap arrays of slightly offset coherent focal fields whose interference modulates the intensity across the sample. For a given degree of parallelization and power, SMX increases the two- and three-photon excited signal of parallelized nonlinear microscopy by a factor of up to 1.5 and 2.5, respectively. To some extent, sensitive regions may be spared out, whereas in regions with weaker nonlinear susceptibilities the intensity is increased. SMX is relevant to all modes of nonlinear microscopy, including parallelized second- and third-harmonic imaging, coherent anti-Stokes Raman scattering, and wide field multiphoton excitation. PMID- 11422667 TI - GTG banding pattern on human metaphase chromosomes revealed by high resolution atomic-force microscopy. AB - Surface topography of human metaphase chromosomes following GTG banding was examined using high resolution atomic force microscopy (AFM). Although using a completely different imaging mechanism, which is based on the mechanical interaction of a probe tip with the chromosome, the observed banding pattern is comparable to results from light microscopy and a karyotype of the AFM imaged metaphase spread can be generated. The AFM imaging process was performed on a normal 2n = 46, XX karyotype and on a 2n = 46, XY, t(2;15)(q23;q15) karyotype as an example of a translocation of chromosomal bands. PMID- 11422668 TI - Cylindrical diameters method for calibrating section thickness in serial electron microscopy. AB - An estimate of section thickness is required for measuring structures in serial section microscopy. Mean section thickness is estimated reliably by averaging the ratios of the diameters of cylindrical objects, such as mitochondria, to the number of sections they span. This cylindrical diameters method improves the accuracy of section thickness as inferred from the colour of sections floating in water. The cylindrical diameters method gives the same answer as that obtained by the minimal folds METHOD: It is preferable because it can be done in a series that has no folds that can distort and obscure the objects that are being measured. PMID- 11422669 TI - Stereological estimation of eye volume using the Pappus method. AB - The estimation of object volume from rotationally randomised sections relies on an ancient geometrical principle due to the Alexandrian mathematician Pappus. In this paper we describe two studies that make estimates of eye volume using a stereological version of the Pappus principle. The first study uses a design based version of the Pappus theorem to make estimates of eye volume in Sprague Dawley rats and the second uses a model-based implementation for estimating eye volume in juvenile Dover sole. In the sole study we compare the estimates of volume with estimates made using the Cavalieri method and show that the Pappus method gives identical volume estimates with a markedly lower coefficient of variation (2.5%) than the Cavalieri method (13.5%). PMID- 11422670 TI - Imaging of three-dimensional objects in emission electron microscopy. AB - Under investigation by emission electron microscopy, the shape and size of three dimensional objects are distorted because of the appearance of a characteristic potential relief and a possible contact potential difference between the particles and the substrate. An estimation of these effects for spherical particles is made. It is shown that the apparent size of particles observed in an emission electron microscope (EEM) could be increased as well as decreased depending on the relation between the work functions of the particle and the substrate. The corresponding formulae are given and several possibilities are shown which permit us to determine from the EEM image the real size of particles and their work function relative to the substrate. PMID- 11422671 TI - Quantitative analyses of the biomineralization of different hard tissues. AB - The primary crystallites of the different developing hard tissues have an apatite structure. However, they have crystal lattice distortions representing an intermediate state between amorphous and fully crystalline. We have applied energy-filtering transmission electron microscopy in the selected area electron diffraction mode to analyse different stages of crystal formation in dentine, bone, enamel and inorganic apatite mineral. We have obtained quantitative information on the degree of crystal lattice distortion using the paracrystal theory of Hosemann and Bagchi. We have found that the early formed crystallites of the hard tissues being analysed have a paracrystalline character comparable to biopolymers. However, with maturation, the lattice fluctuations of the crystallites of the hard tissues bone, enamel and dentine decrease to form a typical (para)crystalline character. Also the decrease of the organic proportion in the matrix corresponds to the decrease of the lattice fluctuation of the crystallites in the different hard tissues during maturation. PMID- 11422672 TI - A nanometre-scale non-periodic structural variation in high temperature superconducting ceramics and the implications for properties. AB - Non-periodic structural variation has been found in the high Tc cuprates, YBa2Cu3O7-x and Hg0.67Pb0.33Ba2Ca2Cu3O8+delta, by image analysis of high resolution transmission electron microscope (HRTEM) images. We use two methods for analysis of the HRTEM images. The first method is a means for measuring the bending of lattice fringes at twin planes. The second method is a low-pass filter technique which enhances information contained by diffuse-scattered electrons and reveals what appears to be an interference effect between domains of differing lattice parameter in the top and bottom of the thin foil. We believe that these methods of image analysis could be usefully applied to the many thousands of HRTEM images that have been collected by other workers in the high temperature superconductor field. This work provides direct structural evidence for phase separation in high Tc cuprates, and gives support to recent stripes models that have been proposed to explain various angle resolved photoelectron spectroscopy and nuclear magnetic resonance data. We believe that the structural variation is a response to an opening of an electronic solubility gap where holes are not uniformly distributed in the material but are confined to metallic stripes. Optimum doping may occur as a consequence of the diffuse boundaries between stripes which arise from spinodal decomposition. Theoretical ideas about the high Tc cuprates which treat the cuprates as homogeneous may need to be modified in order to take account of this type of structural variation. PMID- 11422673 TI - Micromechanical and structural properties of a pennate diatom investigated by atomic force microscopy. AB - The mechanisms behind natural nanofabrication of highly structured silicas are increasingly being investigated. We have explored the use of a standard Nanoscope III Multimode atomic force microscope (AFM) to study the silica shell of diatoms. The delicate structures of the shell surface of the diatom Navicula pelliculosa (Breb.) Hilse were imaged and the shell's micromechanical properties were measured semi-quantitatively with a resolution down to approximately 10 nm. The technique to measure elasticity and hardness with the AFM was demonstrated to be useable even on these hard glass-like surfaces. Different experimental configurations and evaluation methods were tested. They gave a consistent result of the shell micromechanical properties. The first results showed that the diatom shell's overall hardness and elasticity was similar to that of known silicas. However, regions with different mechanical properties were distinguished. The elastic modulus varied from 7 to 20 GPa, from 20 to 100 GPa and from 30 to hundreds of GPa depending on the location. In general, the hardness measurements showed similar spatial differences. The hardness values ranged from 1 to 12 GPa but one specific part of the shell was even harder. Hence, certain localized regions of the shell were significantly harder or more elastic. These regions coincide with known characteristic features and mechanisms appearing at the different stages of the shell's growth. These results show that this method serves as a complementary tool in the study of silica biomineralization, and can detect eventual crystalline phases. PMID- 11422674 TI - The use of the transmandibular implant system in extreme atrophy of the mandible; a retrospective study of the results in two different hospital situations. AB - This retrospective study aims to investigate and compare the results obtained with the transmandibular implant (TMI) in two different hospital situations. Group I were patients treated in a teaching hospital since 1984 by a varying group of surgeons and prosthodontists with a varying degree of experience with the TMI. Group II were patients treated in 1993 and 1994 by experienced surgeons and prosthodontists in the hospital where the TMI was originally developed. In group I, a relatively high frequency of reversible (45.4%) and irreversible--i.e. implant loss--(15.2%) complications occurred. In group II, no implants were lost and reversible complications were reported in 10.8% of the patients. In general, the patients of both groups were quite satisfied with the result of the treatment. In both groups, limited bone loss around the implant posts could be observed. An increase in the height of the mandibular bone distal of the lateral posts was frequently observed, especially in group II. It is concluded that both very favourable and less favourable results can be obtained with the TMI. In this study the best results are found in a highly specialized TMI centre, where patients are treated--with only uncompromised implant-posts--by a small group of experienced surgeons and prosthodontics with accurate application of the existing guidelines. PMID- 11422675 TI - Is the superior belly of the lateral pterygoid primarily a stabilizer? An EMG study. AB - The aim of the present study was to compare the activity levels within the two bellies of the lateral pterygoid muscle between different jaw positions to test the hypothesis that the upper head is primarily a stabilizer. Electromyographic (EMG) recordings, using monopolar concentric needle electrodes, were made from 14 healthy subjects during mandibular rest position (RP), clenching in intercuspal position and jaw opening, first about 10 mm and then about 25 mm. Both bellies had very little activity during RP. The activity level of the superior belly was high during clenching and large opening (LO) with a dip during low opening degree. This pattern differed from that of the inferior belly where the activity was relatively low during clenching and then gradually increased to its highest level during LO. The results support that the lower belly is primarily a jaw opener while the superior belly acts as a stabilizer keeping the disc and condyle in a functionally stable position during clenching and jaw movements. PMID- 11422676 TI - The distribution of cyclooxygenase-1 in human temporomandibular joint samples: an immunohistochemical study. AB - Cyclooxygenase-1,2 (COX-1,2) or prostaglandin (PG) H synthase, is the first enzyme of the pathway in which arachidonic acid is oxidized to PGs. Thus, we examined the expression of COX-1 in 16 human temporomandibular joint (TMJ) samples with internal derangement and in 10 control specimens by an immunohistological technique using paraffin-embedded tissue and specific antihuman COX-1 polyclonal antibody. There was obvious distinction of COX-1 immunoreactivity between the control specimens and internal derangement cases, at the endothelial cells and fibroblasts, in the region of posterior and/or anterior loose connective tissues and synovial membrane. The findings of the present study suggest that COX-1 might be an important mechanism for maintaining normal homeostasis at the endothelial cells and fibroblasts with internal derangement of TMJ. PMID- 11422677 TI - Numerical investigation of the factors affecting interfacial stresses in an MOD restored tooth by auto-meshed finite element method. AB - Many researches have addressed the high correlation between the fracture of restored teeth and the prepared cavity geometry. In addition, concerns about bonding versus debonding dental materials from cavity walls and different occlusal force conditions could also alter the mechanical responses in a restored tooth. This study employed an automatic mesh procedure to investigate the mechanical interactions between different interfacial conditions and cavity parameters such as pulpal wall depth under different chewing functions. The results indicated that when occlusal force was applied directly on the tooth, it could increase unfavourable stress dramatically. When interfacial fixation was simulated as the contact condition between the tooth tissue and restorative material, it might increase the fracture potential exponentially compared with the bonded interface. For pulpal wall depth analyses, greater risks of fracture for the remaining tooth were observed in deeper cavity of mesio-occlusal-distal (MOD) restorations and the existence of a pulpal wall is essential even it is only 1 mm above the gingival wall. PMID- 11422678 TI - Candida albicans colonization on thermal cycled maxillofacial polymeric materials in vitro. AB - In the present study, the colonization of a single isolate of Candida albicans on saliva-, serum-coated or protein free (uncoated), thermocycled (4-70 degrees C for 1 min, respectively; 0, 1000 and 10 000 times) 15 commercial maxillofacial materials were investigated, by adenosine triphosphate (ATP) analysis. In the case of control specimens (not thermocycled and uncoated), the fungal colonization appeared to depend upon the type of commercial products used. Thus, the lowest colonization was observed with additional silicone materials, soft acrylic liners with visible light curing, except for one product, whereas visible light curing liners comprising of single paste or single gel exhibited the highest colonization capacity, and cold cured acrylic liners exhibited the intermediate. However, the fungal colonization on the materials was significantly promoted both by thermal cycling (ANOVA; P < 0.01) and a layer of protein coating (saliva, P < 0.01; serum, P < 0.01). When the inter-relation between the fungal colonization and the surface hydrophobicity of the materials were analysed, fungal colonization on 1000- and 10 000-thermocycled materials correlated well with the contact angles of the materials (Student's t-test, P < 0.01), being consistent with the thermodynamic theory. These results, taken together, suggest that the ageing of the materials and the biological fluids of the host promote yeast colonization on maxillofacial materials. PMID- 11422679 TI - Pain measurement in TMD patients: evaluation of precision and sensitivity of different scales. AB - The aim of this study was to evaluate the precision and sensitivity of four different pain rating scales in 59 temporomandibular disorders (TMD) patients. The capacity of describing changes in symptoms during treatment was also addressed. All patients were asked to answer four pain scales: Visual Analogue Scale (VAS), Numerical Scale (NS), Behaviour Rating Scale (BRS) and Verbal Scale (VS). Two measurements were taken before any treatment. After beginning it, follow-ups and repeated measurements were taken 1 week, 15 days, and every month for 6 months. Statistical analysis showed significant differences (P < 0.05) for all scales, but the NS (P > 0.05), when the two initial measurements were analysed. Regarding the sensitivity, all pain scales demonstrated general symptom improvement of 30-50%, when initial and final figures were compared (P < 0.01). Also, the most significant improvement occurred in the first 2 months after beginning the management programme. Authors concluded that the NS was more accurate to measure reproducibility of pain. As for the capacity of expressing changes during the treatment, all scales demonstrated symptom decrease of 30-50% in a period of 6 months. Caution when analysing the results is recommended because of the subjective aspect of pain measurement, the absence of a 'gold standard' for comparison and the natural fluctuation of TMD symptoms. PMID- 11422680 TI - In vitro assessment of corrosive properties of titanium as a biomaterial. AB - Titanium (Ti) is thought to be a highly biocompatible material, and its clinical applications are becoming increasingly frequent. However, there have recently been some clinical papers reporting hypersensitivity and allergic reactions to Ti. The purpose of this study was to assess the corrosive properties of Ti in the intra-oral environment in vitro. Cast pure Ti specimens were immersed in artificial saliva, physiological saline solution, and 128 mmol x L(-1) of lactic, formic and acetic acids for 3 weeks at 37 degrees C with shaking. The colour, weight, surface morphologies and chemical binding state of specimens were observed before and after immersion. Marked discoloration was recognized on the surface of specimens immersed in formic acid, and a significant difference was found between the immersion solutions. Weight changes also varied with solutions; a tendency to increase in formic acid and to decrease in lactic acid. A slight loss was observed in specimens immersed in lactic acid and artificial saliva. The oxide layer composed mainly of Ti-oxide on the surface of the immersed specimens was thinnest in lactic acid, and thickest in formic acid. The present study indicates that both hydrogen evolution type and oxygen diffusion type corrosion on Ti surfaces are possible in the living body. (Our results also confirm some clinical studies reporting Ti accumulation in surrounding tissues and Ti causing allergic reactions.) PMID- 11422681 TI - Effect of curing with a plasma light on the properties of polymerizable dental restorative materials. AB - Specimens of light-curable dental restoratives have been prepared using either a conventional dental curing lamp (for 20 or 30 s) or a plasma light (for 1 or 2 s). The specimens were then stored in water until their mass equilibrated, then dried to constant mass. Most specimens lost material in this process but the losses in all specimens cured with the plasma light were significantly greater than those cured with the conventional lights (P < 0.05). Longer cure times gave slightly reduced losses in water in most cases. The specimens were then returned to water and allowed to re-equilibrate and their equilibrium water uptake determined. There was no simple trend in this latter property because elution of loosely bound hydrophilic species may have resulted in a less hydrophilic specimen, whose equilibrium water content was therefore correspondingly lower. Overall, the losses through dissolution in water suggest that plasma curing is less effective for these materials than conventional light curing, as it probably results in material with lower molar mass. The losses for the resin-modified glass-ionomer were much greater than for other materials, and it was concluded that the more rapid polymerization with plasma light caused a significant inhibitation of the acid-base part of the setting process. These findings suggest that long-term durability of materials may be compromised by employing plasma light cure rather than a conventional cure system and further studies of this point are recommended. PMID- 11422682 TI - A histomorphometric analysis on bone dynamics in denture supporting tissue under continuous pressure in streptozotocin-induced diabetic rat. AB - The purpose of this study was to investigate bone dynamics in denture supporting tissue under continuous pressure in the diabetic condition by using bone histomorphometry in relation to initial intensity of continuous pressure exerted through the denture base. The experimental denture base, which was designed to load initial continuous pressure of 0.0, 1.0, 10.0 or 20.0 kPa to the denture supporting tissue, was applied to the molar region of hard palate of streptozotocin-induced diabetic rats. Fluorescent labelled palatal bone tissue was stained with Villanueva bone stain and was prepared for the undecalcified grinding section. In 0.0 kPa group, no bone resorption was observed and bone formation was transiently inhibited after the denture insertion. In 1.0 kPa group, although no bone resorption was observed, the beginning of bone formation after the inhibition of bone formation was later than that in 0.0 kPa group and bone formation dynamics after the resumption of bone formation was similar to that in 0.0 kPa group. In 10.0 and 20.0 kPa groups, bone resorption was observed until 3 and 4 weeks, respectively, and the amount of bone resorption for each group was 60 +/- 16 and 87 +/- 18 microm, respectively. The resumption of bone formation in 10.0 and 20.0 kPa groups were observed at the same stage with 1.0 kPa group, and the bone formation dynamics after the resumption of bone formation in 10.0 and 20.0 kPa groups were also similar to that in 0.0 kPa group. From the results of this study, it was revealed that bone formation following bone resorption did not cause equivalent recovery of the bone surface level to the level observed in the case without bone resorption in the diabetic condition. PMID- 11422683 TI - Variation of heads of lateral pterygoid muscle and morphology of articular disc of human temporomandibular joint--anatomical and histological analysis. AB - To grasp the documented evidences available regarding the cause of the anteriorly displaced human temporomandibular joint (TMJ) disc. With 20 specimens of human TMJ from cadavers, at first the number of heads of lateral pterygoid muscle (LPM) bundles were analysed with macroanatomical method. Secondary histological procedure with haematoxyline and eosine staining and orceine staining were carried out for the same specimens. Macroscopically, in 13 specimens an upper and a lower head of the LPM were present. In the other seven specimens, not only an upper and a lower head, but also a third inner head of the LPM covered with a separate fascia was observed. Histochemically, the specific localization of certain bundles of thick elastic fibres in the antero-medial parts of the anteriorly displaced deformed TMJ disc were observed. PMID- 11422684 TI - Prediction of demand for treatment of temporomandibular disorders. AB - Baseline (BL) data on signs and symptoms characteristic of temporomandibular disorders (TMDs) were analysed in 67 young non-patients participating as controls in a clinical trial. Data on the incidence of demand for treatment of TMD were collected during the 4-year follow-up period, and, by means of a mailed questionnaire, during an additional 2 years. The variables analysed included palpation and auscultation of the joints, palpation of the muscles of mastication, experience of joint sounds, of pain on jaw movements, of ear symptoms, awareness of a clenching habit, and frequency of headache. A present absent dichotomy was applied throughout. Palpation tenderness of masticatory muscles was significantly (P=0.02) more often present at BL in those who subsequently requested treatment than in those who did not. The same was true of the combination of palpatory tenderness and clenching habit (P=0.007), whereas the clenching habit alone failed to show a significant difference (P=0.06). However, the predictive values remained too low to be clinically useful, with a possible exception of the negative predictive value of muscle tenderness. The findings indicate that the variables conventionally used to describe the functional status of the masticatory system are at best of modest value in the estimation of individual risk of TMD in healthy young subjects. PMID- 11422685 TI - Multifactorial analysis of an MOD restored human premolar using auto-mesh finite element approach. AB - The use of an mesial-occlusal-distal (MOD) restoration in repairing a large carious lesion depends on many factors. Biomechanical performance is one of the most important. It has been recognized that resistance to restoration failure is not solely a biological concern (e.g. toxicity), but that the cavity shape, dimensions, and the state of stress must all be taken into account. In the present study, a newly developed auto-mesh program was used to generate 30 three dimensional (3D) finite element (FE) models simulating the biomechanics for multiple factorial design of the MOD gold restoration in a maxillary second premolar. Stress levels were related to individual design factors (e.g. pulpal wall depth [P], isthmus width [W] and interaxial thickness [T]) and to their interactions under the worst physiological scenario: a concentrated bite force acting on lingual cusp with debonded interfaces between cavity walls and restorations. The results showed that enlarging the volume of the MOD cavity significantly increased stresses in enamel but did not intentionally affect stresses in dentin. The alternation of individual design parameters significantly changed the peak stresses (P < 0.05). For all three parameters, except for the width, the peak stress increased as the cavity dimension increased. Stress elevation rate (termed as 'volumetric stress rate'--stress elevation by increasing one unit volume of the restored materials) was different among three design factors. Depth was the most critical factor governing the stress elevation in enamel (1.76 MPa mm(-3)) while length (interaxial thickness) was the most important parameter in dentin (0.49 MPa mm(-3)). Width was the least compromising factor to the remaining tooth, 0.32 MPa mm(-3) for enamel and -0.23 MPa mm(-3) for dentin. The findings, at its core, did not fully agree with the traditional concept that the preservation of tooth substances will reduce risk of tooth fracture. This study leaves open possibility for the structural optimization of the MOD restoration. PMID- 11422686 TI - Degree of conversion and temperature rise during polymerization of composite resin samples with blue diodes. AB - To ensure an adequate clinical composite filling light source for photopolymerization is of great importance. In everyday clinical conditions commonly used unit for polymerization of composite material is halogen curing unit. The development of new blue superbright light emitting diodes (LED) of 470 nm wavelengths comes as an alternative to standard halogen curing unit of 450-470 nm wavelengths. The purpose of this study was to compare the degree of conversion (DC) and temperature rise of four hybrid composite materials: Tetric Ceram, Pertac II, Valux Plus and Degufill Mineral during 40 s illumination with standard halogen curing unit Heliolux GTE of 600 mW cm(-2) intensity, Elipar Highlight soft-start curing unit of 100 mW cm(-2) (10 s) and 700 mW cm(-2) (30 s) intensity and 16 blue superbright LED of minimal intensity of 12 mW cm(-2) on the surface and 1 mm depth. The results revealed only a little bit higher DC values in case of polymerization with even 66 times stronger halogen curing units which showed twice higher temperature than blue diodes. Temperature and DC obtained are higher on the surface than on 1 mm depth regardless on the light source used. PMID- 11422687 TI - Comparison of maximum bite force and dentate status between healthy and frail elderly persons. AB - The purpose of the present study was to (1) determine the standard value of maximum bite force and to (2) compare the maximum bite force of the elderly between healthy and frail subjects. Subjects included 349 healthy elderly individuals (149 males, 200 females) and 24 frail elderly individuals (seven males, 17 females) ranging from 65 to 74 years of age. Maximum bite force was evaluated using a Dental Prescale system. The maximum bite force of the healthy subjects was significantly higher than that of the frail subjects in both males (P=0.020) and females (P=0.015). However, no significant difference was observed in the number of present teeth between the healthy and frail subjects. Median of maximum bite force in healthy males was 408.0 N, and that of the healthy females was 243.5 N. These results suggest that the frail elderly have latent bite force problems. PMID- 11422688 TI - Analysis of stiffness and stress in I-bar clasps. AB - An I-bar clasp is one of the most popular direct retainers for distal-extension removable partial dentures. However, no adequate information is available on the shape associated with biomechanics. This study aimed (1) to establish a three dimensional (3D) finite-element modelling method of I-bar clasps, and (2) to clarify the effect of the shape on the stress and stiffness of I-bar clasps. 3D computer models of I-bar clasps were created with vertical and horizontal straight sections connected with a curved section with six parameters: thickness of the clasp tip (T), width of the clasp tip (W), radius of the curvature (R), horizontal distance between the base and the vertical axis (H), vertical dimension between the tip and the horizontal axis (V), taper (change of width per unit length along the axis)(Tp). Stress decreased as T, W, R and Tp increased, and as V decreased. Stiffness (which is proportional to retention) increased as T, W, R and Tp increased, and as H and V decreased. In both stress and stiffness, the effects of T and Tp were especially large. From the results, a systematic formula between the clasp shape and the stiffness was derived. PMID- 11422689 TI - The pattern of toothwear amongst 12-18-year-old students in a Nigerian population. AB - The purpose of this study was to investigate the pattern of toothwear in the permanent teeth of 12-18-year-old Nigerian students. The study group comprised 176 individuals (67 males and 109 females) who had no carious cavity, restoration or fracture on any of their permanent teeth. Within the group, 86% had tooth surface loss that was limited to enamel and dentine. No statistically significant differences were observed between the sexes. The most commonly affected tooth was the lower first molar, while the occlusal surface was the most frequently involved. One student had toothwear with a predominantly erosive component as a result of frequent ingestion of low pH drinks, while another displayed toothbrush abrasion. Of those with toothwear, two (13.4%) had parafunctional activities. The cause of toothwear in 73.2% of the study population could not be ascertained. Some of it may be connected with the coarse, abrasive diets that many of the students admitted to consuming on a regular basis. PMID- 11422690 TI - Denture base polymer Alldent Sinomer: mechanical properties, water sorption and release of residual compounds. AB - The aim of this study was to determine flexural properties, water sorption, solubility and release of residual compounds of a new paste type denture base polymer Alldent Sinomer. In addition, an effort was made to reinforce the denture base polymer with a polymer-pre-impregnated glass fibre reinforcement (Stick). Six rhombic test specimens were fabricated for the flexural test in accordance to the ISO 1567 standard. Water sorption and solubility was also determined as described in the ISO standard. Residual compound release into water was determined with the high performance liquid chromatography (HPLC) METHOD: The flexural strength of Sinomer polymer was 85.8 MPa and flexural modulus 2.73 GPa. Storing of test specimens in water reduced the flexural strength and the modulus. By adding the glass fibre reinforcement into the test specimen, the flexural strength and the modulus increased considerably (ANOVA, P < 0.001). The increase was minor with the reinforcement of low fibre content and high with the reinforcement of higher fibre content. Water sorption of the Sinomer polymer was 26.4 microgram mm-3 and solubility was 1.52 microgram mm-3. High performance liquid chromatography analysis revealed the release of 0.18 wt% fraction of compound which was determined as methylmethacrylate (MMA) monomer. The results of this study suggest that the new denture base polymer Sinomer passed the requirements of the ISO standards and that its mechanical properties can be increased considerably by an incorporation of a glass fibre reinforcement. The release of MMA monomer into water from the polymer should be taken into account when the polymer is used in denture bases of patients with an allergy towards MMA. PMID- 11422691 TI - "The first bite of the cherry": Intra-oral manipulation prior to the first bite in humans. AB - The breakdown of food in the mouth during mastication can be described in terms of two parameters: a breakage function, which describes the fragmentation of food after a bite, and a selection function which defines the probability of particle fracture. The non-zero value of the selection function depends on the manipulation of food particles by the tongue. Little, however, is known about this. As a first step, this study investigated the manipulation of wax sheets of differing sizes and shapes by the tongue after ingestion. It was found that subjects tended to orientate rectangular and square wax wafers so that the long axis of the particle was parallel to the tooth row, independent of the initial orientation given when they were introduced into the mouth. Circular wafers were randomly oriented relative to initial orientation. If this could be extrapolated to the start of mastication, then it suggests that the tongue tends to align food particles so that the post-canines produce close to the greatest surface area possible by fragmenting them along their longest axis. PMID- 11422692 TI - Curing depth of different shades of a photo-activated prosthetic composite material. AB - This study determined the depth of cure of different shades of a prosthetic composite material with the aim of evaluating the influence of shade variation on post-curing material properties. Four light shades having small tabs (A1, B1, C1 and D2) and four dark shades having higher tabs (A4, B4, C4 and D4) of a prosthetic composite (Artglass) for body paste based on the Vita Lumin Shade guide were selected. Specimens of each shade were exposed with the proprietary photo-curing unit (UniXS) for periods 20, 30, 60 and 90 s. The curing depth of the material for each shade was determined with a scraping technique described by the International Organization for Standardization (ISO 4049), and average values of groups of five specimens were compared using analysis of variance (ANOVA) and Scheffe's S intervals (P < 0.05). The L*a*b* colour parameters of five specimens after 90 s exposure were measured using a small-area dental colorimeter (ShadeEye) in order to determine the colorimetric differences. Three-factor ANOVA revealed that the depth of cure was influenced by shade letter (A, B, C or D) and shade tab (1 and 2, or 4) as well as by the exposure period (P=0.05). Curing depth of the light shades was consistently greater than that of the dark shades. Among the eight shades selected, B1 shade demonstrated the greatest curing depth, while A4 shade exhibited the lowest curing depth. For all shades, longer exposure increased the depth of cure. All of the light shades exhibited higher L* values than any of the four dark shades. Curing depth of the composite material was found to be related to the Vita shade variation and the exposure period. PMID- 11422693 TI - Oral habits among adolescent girls and their association with symptoms of temporomandibular disorders. AB - AIM: To evaluate the contribution of certain parafunctional activities to the presence of temporomandibular disorder (TMD) symptoms among teenage girls, with special emphasis to gum chewing and jaw play. METHODS: A total of 323 girls, aged 15-16 years, were randomly selected from a religious junior high school. The girls responded to a questionnaire on oral habits and TMD symptoms. RESULTS: Gum chewing was a very prevalent habit (62.4%), performed daily (mean chewing time 3.95 h day-1). Girls who chewed intensively (more than 4 h day-1) showed associations with pain in the ear area during function and at rest, as well as with joint noises. Jaw play, although reported by only 14.3%, was significantly associated with pain in the ear area during function and at rest, feeling of tiredness of the jaw while chewing, joint noises, catch and lock. All oral parafunctions, except chewing gum, were associated with jaw play and with each other. CONCLUSIONS: Jaw play was the most detrimental habit in TMD; intensive gum chewing was a potentially contributing factor for joint noises and pain. Oral parafunctions (except chewing gum) were significantly associated between themselves and suggest a behavioural pattern of "jaw hyperactivity". PMID- 11422694 TI - Gingival blood flow measurement with a non-contact laser flowmeter. AB - A non-contact laser flowmeter was used to measure the changing of the gingival blood flow. Five university students with healthy oral condition were selected in this study. The blood flow measurement on the extensor digitorum (above the head of third metacarpal), with the changing of distance and angle between the probe and the tissue was used as a pre-study experiment. Blood flow rate was determined in the labial gingiva (2 mm above the cervical line) of upper central incisor using a stent fixing the probe at a 3-mm distance from the tissue. A basal level of gingival blood flow was taken two times each day for 5 days. The effects of water of different temperatures on the gingival blood flow are discussed. With the changing of distance, the blood flow rate became smaller, but there was no significant effect from the angle. The reproducibility was acceptable through the 5-day measurement. After stimulating with warm and body temperature water, the blood flow first increased significantly and then went back to the basal line (faster with the body temperature water). With cold water, different reactions between the subjects were observed. PMID- 11422695 TI - Long-term clinical evaluation of fracture and pulp injury following glass-ionomer cement or composite resin applied as a base filling in teeth restored with amalgam. AB - The aim of this research was to analyse the long-term clinical behaviour of two dental materials applied as filling under silver amalgam restorations: glass ionomer cement (GIC) and composite resin with adhesive system (CR). In this study, 117 posterior teeth (29 premolars and 88 molars) were selected with carious lesions which resulted in great loss of dentin and cusps with unsupported enamel. After caries removal, cavities were prepared and totally filled with GIC or with CR. In a following visit, new cavities were prepared, leaving the employed filling material as a base and support for the enamel, which were then restored with silver amalgam. Restorations were evaluated periodically after 6 months and up to 5 years. Both fracture and pulpal involvement rates were low. Although differences could be observed in the behaviour of the materials, statistical survival estimation showed that the performances of GIC and CR as filling material were similar. There was a significant association both between kind of tooth (molar or premolar) and long-term survival of the restorations; and between degree of unsupported enamel and the same long-term survival. Our results confirmed that the technique in which GIC or CR are used as filling under silver amalgam restorations is clinically acceptable. PMID- 11422696 TI - Clinical evaluation of a chemically cured hard denture relining material. AB - The purpose of the current study was to evaluate the clinical performance of a chemically cured denture relining material after 1-year service period. A powder liquid material designed for denture base relining (Tokuso Rebase Normal-set, Tokuyama Corp., Tokyo, Japan.) was assessed. Fifty adult patients, who required denture reline treatment, participated in the current project. Adaptation of each denture was examined with a silicone elastomeric material, and the surface to be relined was reduced by means of a rotary cutting instrument, then conditioned with dichloromethane. The relining material was spatulated, poured into the relining area, inserted and adjusted after setting. Each relined denture was subjected to base line evaluation immediately after relining and end evaluation after 1 year. Standardized criteria were used for direct evaluation of the material. After an observation period of 1 year, the rate of the relining cases considered to be in clinically ideal condition (Alfa) was 90% for staining, 90% for discoloration, 94% for peeling, 96% for adaptation and 100% for irritation to oral tissue. Only one case was judged as clinically unacceptable (Charlie), and this was for staining. On the basis of a 1-year observation, the Tokuso Rebase material is considered to be a clinically reliable hard denture relining agent. PMID- 11422697 TI - Surface roughness and wettability of enamel and dentine surfaces prepared with different dental burs. AB - The aim of dental adhesive restorations is to produce a long lasting union between the restoration and the tooth structure. This bond depends on many variables including the geometry of the preparation and the type of bonding agent or luting cement. It is therefore suggested that the topography of the tooth surface may influence the wettability and the bonding quality of adhesive systems. This study measured the surface roughness and wettability of enamel and dentine after preparation with different dental burs. The mesial and distal surfaces of 15 extracted sound human premolar teeth were prepared with a tungsten carbide crown bur, a diamond bur and a tungsten carbide finishing bur and finished in enamel or dentin, respectively. The prepared surfaces were analysed with a surface profilometer and scanning electron microscopy (SEM). The contact angle of distilled water on each of the prepared surfaces was used as the measure of wettability. The differences in average surface roughness (Ra) were significant between the rotary instrument groups, as revealed by a two-way ANOVA test. No differences were detected between enamel and dentine surfaces prepared with the same type of dental bur. The smoothest surfaces were those completed with tungsten carbide finishing burs. The diamond bur preparations were intermediate in the roughness assessment and the tungsten carbide crown burs gave the roughest surfaces. There were no significant differences in the contact angle measurements for the various groups. It was concluded that the surface roughness of enamel and dentine prepared by different rotary instruments had no significant influence on the wettability of distilled water on these surfaces. PMID- 11422698 TI - A study of factors for successful splint capture of anteriorly displaced temporomandibular joint disc with disc repositioning appliance. AB - The purpose of this study was to analyse the factors which influenced the success of disc recapture by the insertion of a disc repositioning appliance. Fifty-one joints with joint clicking that occurred at both middle to late opening and late closing (near maximum cuspation) of the mandible were splinted with a mandibular full-coverage repositioning appliance. The clinical and MR findings were compared between the joints with successful and unsuccessful splint disc capture. Thirty two clicking joints with reducibly displaced discs (DDWR) had successful disc recapture, while six of 19 joints with displaced disc without reduction (DDWOR). Unsuccessful joints with DDWOR had significantly higher prevalence of deformed disc and joint effusion, higher VAS quantitative pain score, and severe disc displacement especially in medial part of the joint (P < 0.05). From the results of this study joints with DDWR can be expected to have successful disc recapture with the insertion of the appliance. In joints with DDWOR, presence of inflammatory conditions, changed disc morphology and extensive disc displacement in medial part of the joint are negative factors. PMID- 11422699 TI - Flexural behaviour of post-cured composites at oral-simulating temperatures. AB - Post-curing treatments have been known to improve the mechanical stability of visible light-cured composites. After individual post-curing treatment, the flexural strength (FS) of four commercial direct/indirect placement composite materials which differ greatly in composition [oligocarbonate dimethacrylate (OCDMA)-based Conquest C & B (CQT), Bisphenol-A glycidyl dimethacrylate (BisGMA) based Charisma, urethane dimethacrylate (UDMA)-based Concept (CCT), and BisGMA/UDMA-based Dentacolor] was evaluated under water in the temperature range of 12-50 degrees C. A control series was tested in air at room temperature (25 +/ 1 degrees C). Data were analysed using ANOVA and Duncan's test. Flexural strengths overall decreased (20-40%, P < 0.01) with increasing temperatures except with Conquest C & B. Surprisingly, higher FS values were found in wet conditions than in dry conditions at 25 degrees C. UDMA-based materials much more easily undergo softening in water and by temperature change than do BisGMA- or OCDMA-based materials. Post-cured composites can be significantly affected by exposure to oral environments. Different composition determines the degree of influence. PMID- 11422700 TI - Oral functional characteristics and gastrointestinal pathology: an epidemiological approach. AB - One of the main problems for aged people is that of having a balanced diet, improved by correct conditioning of the alimentary bolus. Does the masticatory status influence the appearance of gastrointestinal disorders? Such a question justifies the present study. The present work concerned all the patients (211) of eight geriatric institutions, and was completed by a retrospective approach (case control study). The main result brought by the case-control study, showed that poor oral functional characteristics represent a risk for gastrointestinal pathology (age being neutralized; odds ratio (OR): 2.62). This result indicates that the loss of teeth must be compensated by functionally effective dentures. Not only will the patients' physical and psychic health improve but also medical and public health problems will be solved. PMID- 11422701 TI - Influence on force curve exerted by jaw tapping force. AB - The purpose of this study, which made use of visual biofeedback, was to determine how methods of regulating jaw tapping force differed depending on the strength of the tapping, using the force curve as an index. Nine healthy examinees were asked to make 30-35 jaw tapping movements, reproducing the defined target tapping force as accurately as possible. We measured the duration of the tooth contact phase, the time to peak force, the first time derivative of force (peak dF/dt), and the time to peak dF/dt. The results indicated that the duration of the tooth contact phase and the time to peak force increased with the target value (P < 0.01). As the target rose, the peak dF/dt increased significantly (P < 0.01), but the time to peak dF/dt was not significant (P=0.134). We found that the higher the target value, the greater the degree of dependency on feedback information. We also found that both the peak dF/dt and the time to peak dF/dt were determined for each examinee prior to movement. PMID- 11422702 TI - Fracture strength of copy-milled and conventional In-Ceram crowns. AB - The purpose of this study was to compare the fracture resistance of copy-milled and conventional In-Ceram crowns. Four groups of 10 uniform sized all-ceramic anterior crowns were fabricated for this test: (1) In-Ceram Spinell (2) In-Ceram Alumina (3) Celay In-Ceram Spinell, and (4) Celay In-Ceram Alumina crowns. All specimens were cemented on stainless steel master die with resin cement and stored in 37 degrees C water for one day prior to loading into a universal testing machine. Using a steel ball at a crosshead speed of 0.5 mm min-1, the crowns were loaded at 30 degrees C angle until catastrophic failure occurred. Mean fracture strength was analysed and compared. Under the conditions of this study and the materials used, the following conclusions were drawn: 1. The strength of Celay In-Ceram anterior crowns had a slightly higher fracture strength than conventional In-Ceram crowns. 2. In-Ceram Alumina crowns had a significantly higher fracture strength than In-Ceram Spinell crowns in both conventional and copy milling methods. PMID- 11422703 TI - Three-dimensional finite element analysis of the facial skeleton on simulated occlusal loading. AB - Development of predictive models of occlusal loading of the facial skeleton will be of value for prosthetic design in oral rehabilitation. A 3-D finite element (FE) model of a human skull, based on CT scans, was constructed to analyse strain and stress distribution in the facial skeleton caused by simulated occlusal loading. Vertical loads were applied simulating loading of the full maxillary arch and unilateral single point occlusal loading of maxillary molar, pre-molar, canine and incisor sites. Strain and stress regimes from Von Mises (VM) failure criteria and extension and compression diagrams showed even distribution of strain following loading of the full maxillary arch throughout the facial elements. For individual points, the highest VM concentrations were consistently located on the facial aspect several mm above the loading site. Strain trajectories divided into a 'V-shaped' pattern, from the loading point into medial and lateral branches with higher VM values in the medial. As the same load was applied from the posterior to anterior region, VM values increased on all facial areas. Strain patterns were less symmetric and there was an increase in strain in the alveolar arch and around the rim of the nasal cavity. The overall picture of the facial skeleton is of a vertical plate enabling it to withstand occlusal stresses by in-plane loading and bending in its own plane. The most efficient distribution of load was on maxillary full arch loading with the most unfavourable strain concentrations occurring on loading in the anterior region. PMID- 11422704 TI - Histomorphometric analysis on bone dynamics in denture supporting tissue under masticatory pressure in rat. AB - The bone dynamics under denture base has not been clarified sufficiently. The purpose of this study was to evaluate histomorphometrically the bone dynamics in denture supporting tissue with relation to the intensity of the masticatory pressure. The removable denture bases, which exerted four different regulated masticatory pressures (0.0, 1.0, 20.0 or 40.0 kPa), were inserted to the hard palate of the molar region of 22-week-old rats. The palatal tissues were excised from 1 to 12 weeks after denture insertion at intervals of 1 week and embedded in methyl-methacrylate. No bone resorption was observed throughout the experimental period in the 0.0 and 1.0 kPa groups. In the 1.0 kPa group compared with the 0.0 kPa group, bone formation parameters except for bone formation rate significantly increased at 4 weeks, but there was no significant difference between these groups in the other experimental periods. In the 20.0 and 40.0 kPa groups, bone resorption was observed until 3 and 5 weeks, respectively, and the amounts of bone resorption (AoBR) were 45 +/- 25 and 104 +/- 21 microm, respectively. After bone resorption, bone formation parameters in these two groups showed transiently significant decrease compared with the 0.0 kPa group, but were the same time course as the 0.0 kPa group following this decrease. In conclusion, masticatory pressure caused different time courses of the bone dynamics in denture supporting tissue depending on the initial intensity of the pressure. The AoBR responded to the initial intensity of masticatory pressure. Bone formation following bone resorption did not cause equivalent recovery of the bone surface level to the level observed in the case without bone resorption in the present study. PMID- 11422706 TI - Recovery following laparoscopic cholecystectomy in either a 23 hour or an 8 hour facility. AB - Research confirms that laparoscopic cholecystectomy (LC) results in shorter lengths of hospital stay and earlier return to usual activity than the traditional cholecystectomy procedure. Research in this area, however, focuses more on the medical aspects of patient recovery, but very few studies have evaluated how these patients manage their recovery at home or what types of problems they encounter. A total of 28 LC patients were randomly assigned to two groups: (1) 23 h stay (overnight) in a general surgical ward or (2) day procedure unit (DPU) stay. Data was collected by a self-administered Postoperative Symptoms Diary and telephone interview. Results showed no significant difference between the two groups of patients recovery symptoms scores. Problems with mobility, pain and elimination recorded the highest mean scores for both groups of patients. Overnight patients also experienced problems with tiredness and eating. All DPU patients were able to manage their postoperative symptoms, compared to only 44% of patients who had stayed in overnight. Carer assistance was needed with regard to activities of daily living, child care and reassurance. Results showed that with careful selection of patients, LC cases performed as day procedures did not impact at all on the patients' recovery trajectory. PMID- 11422708 TI - Accessing residential care from an acute hospital: can we be more efficient? AB - Hospitalized patients who require admission to residential care are often thought to make prolonged and inappropriate use of hospital resources. There are no Australian data on the factors that contribute to length of hospital stay for such patients. The aim of this study was to determine the timing of critical steps in discharge planning for hospitalized patients who need residential care. We prospectively audited 100 consecutive referrals to an Aged Care Assessment Team (ACAT) from one acute hospital in South Australia. Case notes were examined to determine the timings of critical events in discharge planning. We found 47% of patients were discharged to a nursing home, 16% to a hostel, 11% died, 10% returned home and 16% went to another facility. The average length of hospital stay was 27.2 days, and an average of 8.4 days elapsed before a decision to seek residential care was first recorded. A further 4.5 days elapsed before ACAT referral, 4.6 days before ACAT approval and 9.7 days before a residential care bed became available. We conclude that people admitted to our hospital from the community and who subsequently need residential care, spend 36% of their stay awaiting a residential care bed. Most of their hospital stay has elapsed before residential care is considered necessary and referral and approval processes have been activated. Strategies to reduce length of stay should perhaps focus on the earlier recognition of the need for residential care and accelerated referral and assessment processes. Earlier involvement by social work and occupational therapy should be considered. PMID- 11422710 TI - Question of 'patients' versus 'clients'. AB - Our study showed that of the 300 consecutive people interviewed at the General Hospital at Port-of-Spain, Trinidad, most persons (80%) preferred to be called 'patients' as opposed to 7% choosing the term 'client' and 6% opting for the term 'customer'. This proves our hypothesis that in the developing world people prefer to maintain the doctor-patient relationship and leave the word 'client' and 'customer' for the businessman. PMID- 11422712 TI - Diabetes in general practice: Tongans in Tonga and South Auckland. AB - The aim of this study was to compare the management and treatment outcomes of Tongan diabetic patients in Tonga and South Auckland, New Zealand. This was achieved by comparison of Tongan diabetic patients within clinical audits from a general practice in Tonga (n = 124) with those from participating general practices in South Auckland (n = 168). Our results indicate that some measures of diabetes care and outcomes were similar or even better in Tonga, while some interventions were not available in Tonga. Control of weight, glucose, blood pressure and cholesterol remained difficult in both areas. Severe diabetic foot damage was more common among Tongan patients in Tonga (6.5% vs 1.8%, P < 0.05). This international comparison shows that Tongans in both Tonga and New Zealand remain at high-risk of complications independent of the health system under which care is being delivered. While barriers to implementation may differ in the two settings, improvements in the co-ordination of care are likely to be of benefit in both settings. PMID- 11422714 TI - Retrospective audit of standards and quality in colposcopy services in a district hospital. AB - A retrospective audit to compare the performance of a colposcopy clinic with the standards set by National Health Service Cervical Screening Programme (NHSCSP) was carried out at Caerphilly District Miners Hospital, South Wales, United Kingdom. The study sample size consisted of 150 women who underwent colposcopy and diathermy large loop excision of the transformation zone (LLETZ) for abnormal cytology during January l995 to January 1998. The results showed deficiencies in the areas of communication for clinic appointments for women with high-grade cytology and in information given to general practitioners about test results. The services fell short of the standards in recording of colposcopy findings and grading of lesions. The rest of the parameters audited reached the NHSCSP standards. Recommendations to improve communication and to increase the objectivity of recording colposcopy findings were made. A more practical approach to set standards in areas of communication was suggested. PMID- 11422716 TI - Audit of sedated versus unsedated gastroscopy: do patients notice a difference? AB - Unsedated diagnostic gastroscopy has become widely accepted as a diagnostic procedure which avoids the risk of an anaesthetic. It also provides advantages for patients and hospitals in converting the procedure to an ambulatory care investigation. Patient perception of the procedure can sometimes differ from that of medical and nursing staff. We have decided to report our usual clinical practice by auditing 100 consecutive patients undergoing this procedure in a large rural private hospital. Patient tolerance was analyzed in various categories including degree of comfort, degree of pain, ease of breathing and willingness to repeat the procedure under the same conditions. The perceived comfort rating was compared between the patient, the endoscopist and the endoscopy nurse. A total of 100 consecutive patients were evaluated; 55 chose to be sedated and 45 were unsedated. Of the 100 patients tested, 88% stated they would have the procedure the same way if a repeat procedure was required. There was no significant difference between male/female or sedated/unsedated patients. The most important consideration for patients who chose to have the procedure unsedated was the ability to speak to the endoscopist immediately post-procedure. Patient rating of pain was not significantly different between the sedated and unsedated groups. There was no significant difference in the independent assessment by the endoscopist and the nurse with respect to patient comfort in both the sedated and the unsedated groups. However, their assessment differed significantly from the patients own rating, as endoscopists and gastrointestinal (GI) nurses rated the patient degree of comfort as higher than the patients themselves (P < 0.01 for doctor/patient and nurse/patient score, Student's t test). No complications were reported in either group of patients during the audit. Unsedated diagnostic gastroscopy is perceived to be an acceptable alternative to a sedated procedure by the majority of patients. Patients rate the procedure as more uncomfortable than their health care professionals, but the majority of patients would still have the repeat procedure the same way. PMID- 11422717 TI - Clinical governance and external audit. AB - This paper describes a model of clinical governance that was developed at South Auckland Health during the period 1995 to 2000. Clinical quality and safety are core objectives. A multidisciplinary Clinical Board is responsible for the development and publicising of sound clinical policies together with monitoring the effects of their implementation on quality and safety. The Clinical Board has several committees, including an organization-wide Continuous Quality Improvement Committee to enhance the explicit nature of the quality system in terms of structure, staff awareness and involvement, and to develop the internal audit system. The second stream stems from the Chief Medical Officer and clinical directors in a clinical management sense. The Audit Committee of the Board of Directors covers both clinical and financial audit. The reporting lines back to that committee are described and the role of the external auditor of clinical standards is explained. The aim has been to create a supportive culture where quality initiatives and innovation can flourish, and where the emphasis is not on censure but improvement. PMID- 11422718 TI - Improving the quality of health-care: personal reflections on some opportunities and impediments. AB - There are considerable opportunities for improving quality of health-care but also significant impediments. Identifying, investigating and responding to adverse events in a way that will limit their chance of recurrence is probably the single greatest opportunity for quality improvement. However, this requires a shift from a culture of blame to one that recognises such events as almost always a system failure. A general rule, the shorter the duration of a hospital admission, the safer it will be, but regrettably this is not well understood by the community. The considerable variation in rates of many medical and surgical interventions between private and public patients and between patients living in different localities provides a great opportunity for determining what rates produce the best health outcomes. It is probable that reducing the health problems of the socioeconomically disadvantaged, a major quality issue, will require innovative approaches to delivery of their health-care. PMID- 11422719 TI - Making the best of consumer participation. AB - Organizations that have engaged consumers in their work using traditional patient satisfaction tools have accomplished a number of significant outcomes. However, organizations are now being challenged to move into more meaningful collaborations with consumers regarding evaluating and improving services. This study describes and evaluates the effectiveness of consumer partnerships established to cover a project that lasted for about 4 months. Surveys were conducted with staff and consumers involved in the project. Structured questionnaires were used for both groups and the consumers were invited to focus interviews to explore specific issues. Issues regarding recruitment, preparation and management of consumers are important if satisfactory outcomes are to be realised. Further, staff attitudes and behaviours were examined that indicated staff shared the view that engagement of consumers was a worthwhile exercise. Quality consumer collaborations can bring about organizations being a highly valued community assets. PMID- 11422720 TI - Phenomenon of quality and health-care: snowball or an avalanche? AB - The quality-of-care in health as a phenomenon is developing through small incremental steps. Some suggest this approach is too slow and more rapid change is warranted. It is possible to achieve more rapid change by reaching a 'tipping point' or a critical level. To reach a critical level requires (i) identifying key people, (ii) having an idea that sticks, and (iii) having the right context. Examples from selected aspects of quality in health-care including the use of report cards, recognition and remedial action for adverse events, the substantial reviews of health systems for quality-of-care suggest the critical level has been achieved. However, when the three rules for reaching the 'tipping point' are examined closely, it would seem that much more work is required to transform quality in health-care from a snowball into an avalanche. PMID- 11422721 TI - ACHS Quality Awards 2000. Quality is the way we do business. AB - Hollywood Private Hospital recognized that the use of quality management processes can achieve numerous benefits; however, for this to occur quality must be regarded as normal business practice rather than a separate programme. Therefore, the means of ensuring a quality service must be embedded in the strategic plans of both the organization and individual departments. The Hollywood Private Hospital Executive committed the organization to this approach further building on the 'core values' of the hospital by: integrating quality into the Strategic Planning of the organization; integrating risk management into the existing quality system; further embedding of the core values into the culture of the organisation; introducing systems thinking into the organization; taking a process improvement approach to improving quality; involving staff in Quality Action Teams and utilizing the Evaluation and Quality Improvement Programme as the management framework to co-ordinate all the above. PMID- 11422722 TI - Increased cerebral response during a divided attention task following sleep deprivation. AB - We recently reported that the brain showed greater responsiveness to some cognitive demands following total sleep deprivation (TSD). Specifically, verbal learning led to increased cerebral activation following TSD while arithmetic resulted in decreased activation. Here we report data from a divided attention task that combined verbal learning and arithmetic. Thirteen normal control subjects performed the task while undergoing functional magnetic resonance imaging (FMRI) scans after a normal night of sleep and following 35 h TSD. Behaviourally, subjects showed only modest impairments following TSD. With respect to cerebral activation, the results showed (a) increased activation in the prefrontal cortex and parietal lobes, particularly in the right hemisphere, following TSD, (b) activation in left inferior frontal gyrus correlated with increased subjective sleepiness after TSD, and (c) activation in bilateral parietal lobes correlated with the extent of intact memory performance after TSD. Many of the brain regions showing a greater response after TSD compared with normal sleep are thought to be involved in control of attention. These data imply that the divided attention task required more attentional resources (specifically, performance monitoring and sustained attention) following TSD than after normal sleep. Other neuroimaging results may relate to the verbal learning and/or arithmetic demands of the task. This is the first study to examine divided attention performance after TSD with neuroimaging and supports our previous suggestion that the brain may be more plastic during cognitive performance following TSD than previously thought. PMID- 11422723 TI - Temporal and stagewise distribution of high frequency EEG activity in patients with primary and secondary insomnia and in good sleeper controls. AB - In the present study, we evaluate the temporal and stagewise distribution of high frequency EEG activity (HFA) in primary and secondary insomnia. Three groups (n=9 per group) were compared: primary insomnia (PI), Insomnia secondary to major depression (MDD), and good sleeper controls (GS). Groups were matched for age, sex and body mass. Average spectral profiles were created for each sleep epoch. Grand averages were created for each NREM cycle and each stage of sleep after removing waking and movement epochs and epochs containing micro or miniarousals. It was found that HFA (in terms of relative power) tends to increase across NREM cycles, occurs maximally during stage 1 and during REM sleep, and that both these effects are exaggerated in patients with PI. In addition, HFA was found to be inversely associated with Delta activity and the three groups in our study appear to exhibit characteristic Delta/Beta patterns. Our data are consistent with the perspective that HFA is related to CNS arousal to the extent that Beta/Gamma activity occurs maximally during shallow stages of sleep and maximally in subjects with PI. PMID- 11422724 TI - Age, gender and early morning highway accidents. AB - Accident register data, time budget studies and road traffic flow data were used to compute the age and gender-dependent relative risk [odds ratio (OR)] of being involved in a driving accident in which the driver was injured or killed. Alcohol related accidents were excluded from the analysis. The results showed that the night-time risk, compared with that of the forenoon, was dramatically increased (OR=5) for young drivers (18--24 years) and reduced for old (65+) drivers. In direct comparison, the young drivers had 5-10 times higher risk of being involved in an accident during late night than during the forenoon, with the excess risk during the daytime being considerably lower. Women had a less pronounced night time peak than men. In direct comparison, men had twice as high a risk as women during the late night hours. The results clearly demonstrate a strong effect of young age on night-time accident risk, together with a moderate effect of (male) gender. PMID- 11422725 TI - Is there a link between subjective daytime somnolence and sickness absenteeism? A study in a working population. AB - A number of studies have highlighted the increasing incidence and financial cost of sleep-related disorders in the general population, but little research has been carried out on the impact of subjective daytime somnolence on health status. The existence of a survey of the health of employees of the French National Gas and Electricity Board has allowed us to investigate this question and measure the link between subjective daytime somnolence and sickness absenteeism, used here as a general health indicator. In order to evaluate the quality of sleep over the previous 3 months, a questionnaire was given to each participant. The association between subjective daytime somnolence and absence as a result of sickness was explored using the data of sickness absenteeism provided by the company's social security department during a 12-month follow-up period. Of our 1105 subjects, 6.7% reported severe subjective daytime somnolence of 3 days or more a week and 30% of our study population had at least one spell of sickness absence during the 12-month period of follow-up. A strong association was observed between subjective daytime somnolence and sickness absence, which remained significant even after adjustment for potential confounding variables (age, sex, employment grade, sleep symptoms and self-reported diseases). The odds-ratio for absence as a result of sickness during the follow-up period associated with subjective daytime somnolence of 3 days or more a week was 2.2 (95% CI: 1.3--3.8). Employees suffering from severe subjective daytime somnolence lose more working days for health reasons than their more alert colleagues. This may have long-term implications for employees' health. PMID- 11422726 TI - Impact of attachment styles on dream recall and dream content: a test of the attachment hypothesis of REM sleep. AB - We tested the hypothesis (McNamara 1996; Zborowski and McNamara 1998) that dream recall and dream content would pattern with interpersonal attachment styles. In study I, college student volunteers were assessed on measures of attachment, dream recall, dream content and other psychologic measures. Results showed that participants who were classified as 'high' on an 'insecure attachment' scale were significantly more likely to (a) report a dream, (b) dream 'frequently', and (c) evidence more intense images that contextualize strong emotions in their dreams as compared with participants who scored low on the insecure attachment scale. In study II, 76 community dwelling elderly volunteers completed measures of attachment, and dream recall. Participants whose attachment style was classified as 'preoccupied' were significantly more likely to report a dream and to report dreams with higher mean number of words per dream as compared with participants classified as 'securely' attached or as 'avoidant' or as 'dismissing.' Dream recall was lowest for the avoidant subjects and highest for the preoccupied subjects. These data support the view that rapid eye movement (REM) sleep and/or dreaming function, in part, to promote attachment. PMID- 11422727 TI - SSRI treatment suppresses dream recall frequency but increases subjective dream intensity in normal subjects. AB - Clinical lore and a small number of published studies report that the selective serotonin reuptake inhibitors (SSRIs) intensify dreaming. This study examines the dream effects of paroxetine and fluvoxamine in order to both increase clinical knowledge of these agents and to test an important potential method for probing the relationship between REM sleep neurobiology and dreaming in humans. Fourteen normal, paid volunteers (4 males, 10 females; mean age 27.4 year, range 22--39) free of medical or neuropsychiatric symptoms as well as of psychotropic or sleep affecting drugs completed a 31-day home-based study consisting of: 7 days drug free baseline; 19 days on either 100 mg fluvoxamine (7 Ss) or 20 mg paroxetine (7 Ss) in divided morning and evening doses; and 5 days acute discontinuation. Upon awakening, subjects wrote dream reports, self-scored specific emotions in their reports and rated seven general dream characteristics using 5-point Likert scales. Dream reports were independently scored for bizarreness, movement and number of visual nouns by three judges. REM sleep-related measures were obtained using the Nightcap ambulatory sleep monitor. Mean dream recall frequency decreased during treatment compared with baseline. Dream report length and judge rated bizarreness were greater during acute discontinuation compared with both baseline and treatment and this effect was a result of the fluvoxamine-treated subjects. The subjective intensity of dreaming increased during both treatment and acute discontinuation compared with baseline. Propensity to enter REM sleep was decreased during treatment compared with baseline and acute discontinuation and the intensity of REM sleep increased during acute discontinuation compared with baseline and treatment. The decrease in dream frequency during SSRI treatment may reflect serotonergic REM suppression while the augmented report length and bizarreness during acute SSRI discontinuation may reflect cholinergic rebound from serotonergic suppression. PMID- 11422728 TI - The synchronizing influence of Substantia Innominata on the thalamus of the cat. AB - We examined the stimulating effect of Substantia Innominata pars anterior (SIa), during the waking state, on the 'central' part of the Mediodorsal nucleus of the thalamus (MD), combining electrophysiological and anatomical techniques in restrained, undrugged, unanaesthetized cats. Thalamic MD units were recorded, after electrical stimulation of the Substantia Innominata, at 1 Hz, with a single pulse or short trains of four pulses. Responses were studied by poststimulus histograms. In about 64 of the 84 recorded MD neurones (76%), stimulation of the Substantia Innominata, during the waking state, induced a brief cell excitation, followed first by prolonged inhibition of firing and then by a strong excitatory rebound discharge; after this comes a second sequence of inhibition and excitation, of decreasing amplitude. After stimulation of the Substantia Innominata, the MD units tended to start a repetitive discharge at 4--7 Hz. To investigate the connections of Substantia Innominata cells upon the areas where MD units were recorded we injected horseradish peroxidase wheat germ agglutinin (WGA-HRP), combined with immunohistochemistry for glutamic acid decarboxylase (GAD) and choline acetyl transferase (ChAT). Of the total population of retrogradely labelled cells in the Substantia Innominata 53% were GAD positive while less than 16% were ChAT positive. The GAD positive MD-projecting cells in the Substantia Innominata were triangular to fusiform and small to medium in size. These findings indicate that GABAergic input from the Substantia Innominata may contribute to increasing the hyperpolarizing inhibitory pressure on MD cells in the 'central' part during slow wave sleep (SWS). PMID- 11422729 TI - Between-sleep states transitions in premature babies. AB - To assess the manner in which between-sleep state transitions occur in infants, we examined polysomnography (PSG) studies in 25 clinically and neurologically normal, appropriate-for-gestational-age, 30-to-36-week-gestational age (GA) infants. Twenty infants underwent paper PSG and five infants digitised PSG. Sleep states were coded based on concordance of REMs and the electroencephalogram (EEG) pattern. Data were analysed using a multivariate linear model, with the subject factor as a cluster. Duration of active sleep (AS) to quiet sleep (QS) transitions (median 4.8 min) was significantly longer than duration of QS to AS transitions (1.7 min) and was independent from GA and from the recording method (paper vs. digitized PSG). The sequence of modifications in parameters (REM and EEG) was invariable: REM cessation was the first change in AS to QS transitions, and REM appearance was the last change in the QS to AS transitions. Our study demonstrates a stable, well-organized pattern of between-sleep-states transitions in healthy 30-to-36-week GA premature infants. These findings are similar to those described in full-term newborns and are in agreement with our previous observations of well-defined sleep states at the age investigated here. PMID- 11422730 TI - Decreased sleep quality in patients suffering from retinitis pigmentosa. AB - The purpose of this study was to assess the prevalence of sleep disturbance in subjects diagnosed with retinitis pigmentosa (RP), as well as the influence of age and gender. Sleep quality was assessed, by means of the Pittsburgh Sleep Quality Index (PSQI), in people with RP (n=177) and gender-and age-matched normally sighted individuals (n=491). The population was divided, according to their age, in eight decade groups. People on shift-work, with affective disorders or with visual impairment other than RP, were excluded. The influence of cataracts in sleep quality was also studied in non-RP people (n=57), with cataracts significantly impairing visual acuity. Another group of healthy controls (n=190) was studied in different seasons of the year for a possible seasonality in sleep disturbance. Global sleep quality decreased in an age dependent manner in RP-patients, especially from the second decade of life. Retinitis pigmentosa-patients showed, in relation to age-matched controls: lower subjective sleep quality and efficiency, longer sleep latency, shorter sleep duration, higher daytime dysfunction and a higher use of sleeping medication. No significant differences in sleep quality were found among RP-patients or controls depending either on their gender or on the presence of cataracts. Normal sighted individuals did not show seasonality in their sleep quality. We conclude that the sleep quality of RP-patients decreases in an age-dependent manner and points to the probably degeneration of photoreceptors mediating the photic input to the suprachiasmatic nuclei of the hypothalamus in this disease. PMID- 11422731 TI - Infectious complications of the hemodialysis access. AB - Infectious complications of the vascular access are a major source of morbidity and mortality among hemodialysis (HD) patients. Numerous reports implicate the vascular access in up to 48 to 73% of all bacteremias in HD patients. The incidence of vascular access-related infection is highest when central venous dialysis catheters are employed. Native arteriovenous fistulas carry the lowest risk of infection. Unfortunately, prosthetic arteriovenous grafts, which represent the most common type of HD access in the United States, have been repeatedly shown to be a risk factor for bacteremic and nonbacteremic infections. Silent infection in old nonfunctional clotted prosthetic arteriovenous grafts has recently been recognized as a frequent cause of bacteremia and morbidity among HD patients. High proportions of infections related to the vascular access are caused by staphylococcal organisms, which carry high rates of mortality, recurrence, and metastatic complications. Management of vascular access-related infection has two aspects: The first relates to the choice, duration, and mode of administration of antibiotic therapy. Empiric antibiotic therapy, guided by demographic data and severity of illness, should be employed when the causative organisms are unknown. Prolonged administration of specific parenteral antibiotics is crucial in decreasing complications of infection, especially in cases of staphylococcal bacteremia. The second aspect relates to management of the vascular access. Efforts directed toward bacteriological cure should be concurrent with efforts to preserve native venous access sites whenever possible. Efforts to prevent vascular access-related infection should focus on increasing placement of arteriovenous fistulas and minimizing insertion of central venous dialysis catheters. Careful inspection and monitoring of the vascular access is of paramount importance in early detection of vascular access site-related infections. Several new approaches aimed at preventing catheter and prosthetic graft-related infection are being explored. PMID- 11422732 TI - Peroxisome proliferator-activated receptors (PPARs): novel therapeutic targets in renal disease. AB - Peroxisome proliferator-activated receptors (PPARs): Novel therapeutic targets in renal disease. Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor superfamily of ligand-dependent transcription factors. PPARs play an important role in the general transcriptional control of numerous cellular processes, including lipid metabolism, glucose homeostasis, cell cycle progression, cell differentiation, inflammation and extracellular matrix remodeling. Three PPAR isoforms, designated PPARalpha, PPARbeta and PPARgamma, have been cloned and are differentially expressed in several tissues including the kidney. PPARalpha primary regulates lipid metabolism and modulates inflammation. PPARalpha is the molecular target of the hypolipidemic fibrates including bezafibrate and clofibrate. PPARbeta participates in embryonic development, implantation and bone formation. PPARgamma is a key factor in adipogenesis and also plays an important role in insulin sensitivity, cell cycle regulation and cell differentiation. Antidiabetic thiazolidinediones (TZDs) such as troglitazone and rosiglitazone are specific ligands of PPARgamma, and this interaction is responsible for the insulin-sensitizing and hypoglycemic effect of these drugs. The kidney has been shown to differentially express all PPAR isoforms. PPARalpha is predominantly expressed in proximal tubules and medullary thick ascending limbs, while PPARgamma is expressed in medullary collecting ducts, pelvic urothelium and glomerular mesangial cells. PPARbeta is ubiquitously expressed at low levels in all segments of nephron. Accumulating data has begun to emerge suggesting physiological and pathophysiological roles of PPARs in several tissues including the kidney. The availability of PPAR-selective agonists and antagonists may provide a new approach to modulate the renal response to diseases including glomerulonephritis, glomerulosclerosis and diabetic nephropathy. PMID- 11422733 TI - Nephron endowment in glial cell line-derived neurotrophic factor (GDNF) heterozygous mice. AB - BACKGROUND: The exact molecular mechanisms that regulate ureteric branching morphogenesis in the developing metanephros have not been fully elucidated. However, in vivo and in vitro evidence indicates that glial cell line-derived neurotrophic factor (GDNF) is a key regulator of the initiation of ureteric branching. GDNF knockout mice show renal agenesis or severe dysgenesis and die 24 hours after birth from renal failure. Inhibition of GDNF activity in metanephric organ culture inhibits ureteric branching. Since nephron initiation only occurs at the tips of ureteric branches, the aim of the present study was to determine whether nephron number in GDNF heterozygous mice is reduced. METHODS: Male GDNF heterozygous mice of hybrid 129/Sv and C57/BL genetic background were mated with C57BL/6 females. Offspring were genotyped at postnatal day 30 (PN30) by polymerase chain reaction. Left kidneys were used for estimating kidney volume and total nephron number. We also estimated absolute and relative volumes of ureteric duct epithelium. Unbiased stereological methods were used throughout (Cavalieri method, physical disector/fractionator combination). RESULTS: GDNF wild-type and heterozygous mice had similar body weights at PN30. However, heterozygous kidneys were 25% smaller than wild-type kidneys (wild-type, 114.75 +/- 16.46 mm3; heterozygous, 87.11 +/- 21.84 mm3, P < 0.001) and contained approximately 30% fewer nephrons (wild-type, 11886 +/- 1277; heterozygous, 8573 +/- 2240, P < 0.01). In addition, the absolute ureteric duct volume was significantly reduced in heterozygous mice (P < 0.001). CONCLUSIONS: : These results indicate that the loss of one GDNF allele results in reduced nephron endowment in the adult kidney, presumably as the result of reduced branching morphogenesis of the ureteric bud. PMID- 11422734 TI - Angiogenesis in autosomal-dominant polycystic kidney disease. AB - BACKGROUND: Autosomal-dominant polycystic kidney disease (ADPKD) is a genetic disorder that is responsible for approximately 10% of all cases of end-stage renal disease (ESRD). It is characterized by the formation of epithelial cell cysts, an increase in the extracellullar matrix, and vascular alterations believed to be the result of compression by the cysts. Our recent observations demonstrated a rich vascular network on the surface of the cysts, and thus, we postulated that angiogenesis could be a factor in the progression of ADPKD. METHODS: Kidneys removed from patients with ADPKD were studied using (1) angiographs, (2) immunostaining [factor VIII-related antigen, vascular endothelial growth factor (VEGF), VEGF receptors 1 and 2 (VEGFR-1 and VEGFR-2), metalloproteinase-2 (MMP-2), and integrin alphavbeta3], and (3) Western blot analysis and enzyme-linked immunosorbent assay. The expression of VEGF165 in ADPKD cells in culture was determined. RESULTS: There was (1) an extensive capillary network in the cyst wall of ADPKD kidneys, (2) morphological evidence of vascular malformations, (3) expression of VEGF165 in cyst cells of VEGFR-2 in endothelial cells and an absence of VEGFR-1 in endothelial cells, (4) secretion of VEGF165 by ADPKD cyst cells in culture, and (5) coexpression of matrix MMP-2 and integrin alphavbeta3 in vessels from ADPKD. CONCLUSIONS: There is angiogenesis in ADPKD. This process may be necessary for cyst cells to grow and may be responsible for increased vascular permeability facilitating fluid secretion into the cysts. Neovascularization may result in the formation of aneurysms responsible for the renal bleeding in this disease. PMID- 11422735 TI - Genetic polymorphisms of the renin-angiotensin-aldosterone system in end-stage renal disease. AB - BACKGROUND: Hypertension contributes to the progression to renal failure. A genetic susceptibility to hypertension may predispose to the development of end stage renal disease (ESRD) and promote a more rapid progression to ESRD in patients with renal diseases. Genes encoding for angiotensinogen (AGT), angiotensin-converting enzyme (ACE), and aldosterone synthase (CYP11B2) are candidates for abnormal blood pressure regulation. METHODS: Genotyping was performed in 327 control subjects and 260 ESRD patients for the M235T-AGT, the insertion/deletion (I/D)-ACE, and the -344T/C-CYP11B2 gene polymorphisms using polymerase chain reaction, gel analysis, and appropriate restriction digest when required. RESULTS: Genotype frequencies did not differ significantly between ESRD patients and controls. When ESRD diabetic subjects were compared with diabetic patients without nephropathy, the prevalence of the AGT-MM genotype was lower (28.1 vs. 52.8%, P < 0.01), while the AGT-TT genotype was higher (15.6 vs. 2.7%, P < 0.05). The AGT-TT genotype was associated with a faster progression to ESRD in patients with glomerulonephritis (P < 0.05). In the total ESRD population, progression of renal disease was faster with the ACE-DD than with the DI and II alleles (P < 0.05). This association was particularly strong when the interaction with the AGT genotype was analyzed, with a rapid progression in ACE-DD as compared with ACE-DI and II in patients with the AGT-MM genotype (P < 0.01). CONCLUSIONS: Susceptibility for ESRD and faster progression to ESRD are linked with the AGT genotype in diabetic patients. Faster progression to ESRD is associated with the ACE genotype when the total population with ESRD and with the AGT genotype when patients with glomerulonephritis are considered. Thus, genes of the renin-angiotensin-aldosterone system are candidate genes for further understanding of the interindividual differences in the development and course of ESRD. PMID- 11422736 TI - High glucose induces MCP-1 expression partly via tyrosine kinase-AP-1 pathway in peritoneal mesothelial cells. AB - BACKGROUND: High glucose in peritoneal dialysis solutions has been implicated in the pathogenesis of peritoneal fibrosis in chronic ambulatory peritoneal dialysis (CAPD) patients. However, the mechanisms are not very clear. Peritoneal macrophages seem to participate in the process of peritoneal fibrosis and monocyte chemoattractant protein-1 (MCP-1) plays a key role in the recruitment of monocytes toward the peritoneal cavity. However, little is known about the effect of high glucose on MCP-1 expression and its signal transduction pathway in human peritoneal mesothelial cells. METHODS: Mesothelial cells were cultured with glucose (5 to 100 mmol/L) or mannitol chronically for up to seven days. MCP-1 expression of mRNA and protein was measured by Northern blot analysis and enzyme linked immunosorbent assay (ELISA). Chemotactic activity of high-glucose conditioned culture supernatant was measured by chemotactic assay. To examine the roles of the transcription factors activator protein-1 (AP-1) and nuclear factor kappaB (NF-kappaB), electrophoretic mobility shift assay (EMSA) was performed. RESULTS: Glucose induced MCP-1 mRNA expression in a time- and dose-dependent manner. MCP-1 protein in cell culture supernant was also increased. Equivalent concentrations of mannitol had no significant effect. High-glucose-conditioned supernatant possessed an increased chemotactic activity for monocytes, which was neutralized by anti-MCP-1 antibody. EMSA revealed that glucose increased the AP-1 binding activity in a time- and dose-dependent manner, but not NF-kappaB. Curcumin, an inhibitor of AP-1, dose-dependently suppressed the induction of MCP 1 mRNA by high glucose. Tyrosine kinase inhibitors such as genistein (12.5 to 50 micromol/L) and herbimycin A (0.1 to 1 micromol/L) inhibited the high-glucose induced MCP-1 mRNA expression in a dose-dependent manner, and also suppressed the high-glucose-induced AP-1 binding activity. CONCLUSIONS: : High glucose induced mesothelial MCP-1 expression partly via the tyrosine kinase-AP-1 pathway. PMID- 11422737 TI - Apoptosis and JNK activation are differentially regulated by Fas expression level in renal tubular epithelial cells. AB - BACKGROUND: In chronic renal disease, renal tubular epithelial cell (RTC) Fas expression is up-regulated, leading to apoptotic RTC deletion and tubular atrophy. In vitro, cytokine- or hypoxia-induced up-regulation of Fas expression is associated with RTC apoptosis. In contrast, constitutively expressed, low level RTC Fas does not mediate apoptosis, suggesting that Fas may be coupled to expression level-dependent RTC signaling pathways. Fas is known to signal through JNK in many systems, but the requirement of JNK activation for apoptosis remains controversial. METHODS: To determine if RTC Fas regulates JNK activity and apoptosis, human RTC were transfected with graded concentrations of a eukaryotic expression vector for murine Fas. Apoptosis was measured by annexin V, TUNEL and PARP cleavage assays. JNK activity was determined by immune complex kinase assay and/or immunoblots with phospho-specific JNK antibodies, in the presence or absence of co-expressed dominant negative JNK constructs. RESULTS: Fas antibody stimulation of RTC with high Fas expression levels (to model RTC phenotype in renal disease) caused a tenfold increase in apoptosis, while RTC with low level Fas expression (to model normal RTC phenotype) were apoptosis-resistant. Fas ligation activated JNK in RTC expressing low levels of Fas, but not in apoptosis sensitive RTC with increased Fas expression. Dominant negative JNK co-expression failed to inhibit apoptosis in RTC expressing high levels of Fas, suggesting that JNK is neither necessary, nor sufficient, for Fas-dependent apoptosis. CONCLUSIONS: At high levels of expression, RTC Fas promotes apoptosis in a JNK independent manner. At low basal expression, Fas induces JNK activation, but not apoptosis, consistent with novel roles for RTC Fas as a mediator of cell stress or chronic inflammation. PMID- 11422738 TI - Phosphorylated osteopontin peptides suppress crystallization by inhibiting the growth of calcium oxalate crystals. AB - BACKGROUND: Osteopontin isolated from human urine [uropontin (uOPN)] is a potent inhibitor of calcium oxalate (CaOx) monohydrate (COM) crystallization. However, specific structural features responsible for its effects on CaOx crystallization were not previously known. The present studies were designed to define molecular features responsible for interactions of uOPN with COM crystals and the inhibition of crystallization. METHODS: Peptides and phosphopeptides with sequences corresponding to potential crystal binding domains within the protein sequence of osteopontin were synthesized. Then the effects of these peptides on COM crystal growth and crystal aggregation were investigated and their secondary structures analyzed. RESULTS: Growth of COM crystals was inhibited by approximately 50% at 1000 nmol/L concentrations by the two unmodified peptides with the closest clustering of aspartic acid residues. Growth was not inhibited by the other two unmodified peptides, with aspartic residues more evenly distributed within their sequences. Phosphorylation markedly increased inhibition of COM crystal growth, so that each of the four phosphorylated peptides inhibited growth by at least 50% at concentrations of < or =200 nmol/L. Phosphorylation of these peptides did not cause changes in secondary structure that would favor interaction with COM crystal surfaces. CONCLUSIONS: These studies of synthetic peptides identify molecular features within the osteopontin molecule that contribute to the inhibition of one aspect of COM crystallization. The inhibition of crystal growth induced by phosphorylation appears to result from altered local patterns of charge density, since conformational changes favoring interaction with crystals were not caused by phosphorylation. PMID- 11422739 TI - Angiotensin II infusion increases plasma erythropoietin levels via an angiotensin II type 1 receptor-dependent pathway. AB - BACKGROUND: Angiotensin-converting enzyme inhibitors (ACEIs) have been shown to lower hematocrit and erythropoietin (EPO), but a direct link between angiotensin II (Ang II) and EPO in humans has not been shown. METHODS: Placebo or Ang II was infused for six hours in nine healthy male volunteers with and without blockade of the Ang II subtype 1 receptor (AT1R). EPO concentrations were measured 3, 6, 12, and 24 hours after the start of the infusion. RESULTS: Ang II raised the mean arterial pressure by about 20 mm Hg. Consistent with the known diurnal variation, EPO levels rose significantly (P < or = 0.02) during the day in all groups. During Ang II infusion, EPO levels rose to significantly higher levels after 6 and 12 hours compared with placebo [9.9 +/- 3.5 vs. 7.2 +/- 3.1 mU/mL (3 h, P = NS); 16.9 +/- 4.5 vs. 8.8 +/- 3.7 mU/mL (6 h, P = 0.01); 17.0 +/- 8.6 vs. 11.1 +/ 4.7 mU/mL (12 h, P = 0.01)] and returned to baseline after 24 hours (7.9 +/- 3.8 vs. 10.6 +/- 8.6 mU/mL, P = NS). With AT1R blockade, blood pressure remained normal during Ang II infusion, and EPO levels were never significantly different from placebo [6.8 +/- 4.8, 10.5 +/- 5.6, 13.1 +/- 9.0, and 12.4 +/- 10.1 mU/mL at 3, 6, 12, and 24 h after infusion, respectively, P = NS]. CONCLUSIONS: Ang II increases EPO levels in humans. This increase requires the participation of AT1R. PMID- 11422740 TI - Effects of 1,25(OH)2D3 in experimental mesangial proliferative nephritis in rats. AB - BACKGROUND: 1,25-Dihydroxyvitamin D3 [1,25(OH)2D3], the active form of vitamin D3, is a potent immunomodulatory agent on several cell types such as monocytes and mesangial cells. Recruitment of inflammatory cells, as well as stimulation of resident cells and mesangial matrix accumulation are key features of various experimental and human glomerular diseases. Here we show that 1,25(OH)2D3 attenuates the morphologic and functional alterations in anti-Thy-1.1. nephritis, an experimental model of mesangial proliferative glomerulonephritis. METHODS: The anti-Thy-1.1 group (group I) comprised 24 rats that at day 0 received 0.5 mL of saline containing 400 microg of monoclonal antibodies (mAb) anti-Thy-1.1 IgG. The anti-Thy-1.1 treated with 1,25(OH)2D3 group (group II) were 24 rats given 1,25(OH)2D3 at the dose of 25 ng/100 g body wt/day, from day -3 to day 14. At day 0, the rats received 400 microg of anti-Thy-1.1 monoclonal IgG. The control group (group III) comprised 12 rats injected with vehicle alone; the control group treated with 1,25(OH)2D3 (group IV)-12 rats were given 1,25(OH)2D3 as in group II without mAb administration. Proteinuria and urinary interleukin-6 excretion were measured daily. Blood urea nitrogen and creatinine, creatinine clearance, calcium, and phosphate were measured at days 0, 4, 7, and 14. In addition to conventional periodic acid-Schiff staining, binding of anti-Thy-1.1 IgG and C3b complement fraction, His48- and ED1-positive cells were studied by immunofluorescence. Mesangial proliferation was studied by the proliferating cell nuclear antigen (PCNA) technique. Apoptosis was evaluated by the TUNEL assay. RESULTS: The anti-Thy-1.1 treated with 1,25(OH)2D3 group versus the anti-Thy-1.1 alone group showed a significant reduction in urinary protein (at day 7, 424 +/- 228 vs. 66 +/- 30 mg/mg urinary creatinine, P = 0.02) and interleukin-6 excretion (at day 3, 537 +/- 360 pg/mL vs. 110 +/- 34 pg/mg urinary creatinine, P = 0.015), reduced glomerular diameters (at day 7, 283 +/- 38 vs. 261 +/- 48 microm, P < 0.01), decreased neutrophil (at day 4, 20 +/- 12 His48-positive cells/glomerulus vs. 3.7 +/- 1.3 His48-positive cells/glomerulus, P < 0.001), and monocyte accumulation (day 7, 4.9 +/- 2.9 ED1-positive cells/glomerulus vs. 2.8 +/- 2.9 ED1-positive cells/glomerulus, P < 0.05), and attenuated glomerular cells proliferation (day 7, 13 +/- 3.2 PCNA-positive cells/glomerulus vs. 9.4 +/- 3 PCNA-positive cells/glomerulus, P < 0.01). Apoptosis showed a biphasic behavior with an early peak at day 4 in the anti-Thy-1.1 group (2.3 +/- 2.2 TUNEL-positive cells/glom) related to cellular lysis and a late peak at day 14 related to the recovery phase. CONCLUSIONS: 1,25(OH)2D3 can reduce glomerular hypercellularity, inflammatory infiltration in anti-Thy-1.1 nephritis, preserving the apoptotic response of the reparative phase. PMID- 11422741 TI - Connective tissue growth factor in tubulointerstitial injury of diabetic nephropathy. AB - BACKGROUND: Chronic interstitial fibrosis, which follows the onset of glomerular proteinuria, importantly contributes to progressive renal failure in diabetic nephropathy. The present studies examine the potential role of tubular connective tissue growth factor (CTGF). METHODS: The expression of CTGF was examined in rats with diabetic nephropathy. Regulation and actions of CTGF were studied in in vitro cell culture models. RESULTS: CTGF mRNA levels were increased in the renal cortex of rats with streptozotocin-induced diabetes compared with controls. Immunohistology indicated that CTGF was expressed in renal cortex of diabetic rats, in contrast to controls in some tubular cross-sections, particularly dilated-appearing proximal tubules, in which it tended to colocalize with insulin like growth factor-I (IGF-I). Glomerular ultrafiltrate from diabetic rats, which contained bioactive transforming growth factor-beta (TGF-beta) and hepatocyte growth factor (HGF), induced increased CTGF expression in tubular cells. TGF beta1 and, to a lesser extent, HGF also raised CTGF expression in cultured proximal tubular cells. In contrast, high glucose (25 mmol/L) did not increase the secretion of CTGF. In cultured tubular cells, rhCTGF moderately increased fibronectin but not collagen (Col) type I and type III expression. In NRK-49F renal interstitial fibroblasts, CTGF raised Col alpha1III and thrombospondin-1 levels. CTGF has an IGF-binding domain and binds to IGF-I. In NRK-49F cells, IGF I increased the activity of CTGF towards the expression of Col alpha1III. CONCLUSIONS: CTGF is expressed and regulated downstream from TGF-beta and HGF in proximal tubular cells, is induced by diabetic rat glomerular ultrafiltrate, and has moderate profibrogenic activity in tubular cells and renal interstitial fibroblasts, where its activity is IGF-I dependent. By these means, CTGF may act downstream of TGF-beta and HGF and may contribute to chronic tubulointerstitial fibrosis in diabetic nephropathy. PMID- 11422742 TI - Podocyte injury underlies the progression of focal segmental glomerulosclerosis in the fa/fa Zucker rat. AB - BACKGROUND: The progression of diabetic nephropathy to chronic renal failure is based on the progressive loss of viable nephrons. The manner in which nephrons degenerate in diabetic nephropathy and whether the injury could be transferred from nephron to nephron are insufficiently understood. We studied nephron degeneration in the fa/fa Zucker rat, which is considered to be a model for non insulin-dependent diabetes mellitus. METHODS: Kidneys of fa/fa rats with an established decline of renal function and of fa/+ controls were structurally analyzed by advanced morphological techniques, including serial sectioning, high resolution light microscopy, transmission electron microscopy, cytochemistry, and immunohistochemistry. In addition, tracer studies with ferritin were performed. RESULTS: The degenerative process started in the glomerulus with damage to podocytes, including foot process effacement, pseudocyst formation, and cytoplasmic accumulation of lysosomal granules and lipid droplets. The degeneration of the nephron followed the tuft adhesion-mediated pathway with misdirected filtration from capillaries included in the adhesion toward the interstitium. This was followed by the formation of paraglomerular spaces that extended around the entire glomerulus, as well as via the glomerulotubular junction, to the corresponding tubulointerstitium. This mechanism appeared to play a major role in the progression of the segmental glomerular injury to global sclerosis as well as to the degeneration of the corresponding tubule. CONCLUSIONS: The way a nephron undergoes degeneration in this process assures that the destructive effects remain confined to the initially affected nephron. No evidence for a transfer of the disease from nephron to nephron at the level of the tubulointerstitium was found. Thus, each nephron entering this pathway to degeneration appears to start separately with the same initial injuries at the glomerulus. PMID- 11422743 TI - Adenovirus-mediated transfer of the 39 kD receptor-associated protein increases fibrinolytic capacity. AB - BACKGROUND: The mesothelium has an important role in maintaining an adequate fibrinolytic capacity in the peritoneal cavity and thus in preventing the formation of fibrinous peritoneal adhesions by secreting the fibrinolytic enzyme tissue-type plasminogen activator (t-PA). The fibrinolytic activity of human mesothelial cells (HMCs) is counteracted by rapid uptake of t-PA via the low density lipoprotein receptor-related protein (LRP). The 39 kD receptor-associated protein (RAP) is an inhibitor of binding of t-PA to LRP, but RAP itself is also rapidly degraded via LRP. METHODS: Adenovirus-mediated RAP gene transfer technology was used to evaluate the effect of prolonged overexpression of RAP on t-PA accumulation in conditioned medium of HMCs under basal and inflammatory conditions. RESULTS: Infection of HMCs with a recombinant adenovirus carrying the RAP cDNA resulted within one day in t-PA levels that were maximally twofold to threefold increased as compared with noninfected or adenovirus-beta-galactosidase infected cells. Whereas upon prolonged incubation, t-PA levels in the conditioned medium of uninfected cells leveled off because of rapid uptake and degradation via LRP, t-PA concentrations in the medium of adenovirus-RAP-infected cells continued to increase, reaching fivefold control levels after 72 hours. The increased t-PA accumulation persisted for seven days and then slowly returned to control values over the next few weeks. In contrast, the production of a specific inhibitor of t-PA, plasminogen activator inhibitor-1 (PAI-1), was not affected by adenoviral RAP gene transfer. Northern blotting analysis showed that t-PA, PAI-1, and LRP mRNA concentrations were not changed after adenoviral infection, underlining that the elevated t-PA levels are the result of RAP-blocked uptake and degradation of t-PA rather than increased t-PA synthesis. RAP gene transfer also restored diminished fibrinolytic activity of cytokine-treated mesothelial cells. CONCLUSIONS: Adenovirus-mediated transfer of the RAP gene provides an efficient way of transiently increasing the fibrinolytic capacity of mesothelial cells. PMID- 11422744 TI - Expression of inter-alpha-trypsin inhibitor and tumor necrosis factor-stimulated gene 6 in renal proximal tubular epithelial cells. AB - BACKGROUND: Our previous studies have demonstrated that renal proximal tubular epithelial cells (PTCs) may contribute to renal interstitial fibrosis by the generation of transforming growth factor-beta1 (TGF-beta1). In these in vitro experiments, TGF-beta1 was, however, secreted in its latent form. Plasmin has been implicated as a potential physiological activator of TGF-beta1. The inter alpha-trypsin inhibitor (IalphaI) family of serum protease inhibitors together with tumor necrosis factor-stimulated gene 6 (TSG-6) recently have been implicated in the regulation of this protease pathway. The aim of the current study was to examine PTC synthesis of these proteins and to relate it to alterations of plasmin-protease activity. METHODS: PTCs were grown to confluence and stimulated under serum-free conditions with either interleukin-1beta (IL 1beta) or 25 mmol/L D-glucose. Alterations in IalphaI and TSG-6 generation were detected by Western analysis of both membrane extracts and supernatant samples. Alterations in gene expression were examined by reverse transcription-polymerase chain reaction. The effect of alteration in synthesis of TSG-6 on plasmin activity was determined by quantitating plasmin inhibitory activity of supernatant samples by in vitro calorimetric assay prior to and following TSG-6 immunoprecipitation. RESULTS: The data demonstrate that human PTCs constitutively express mRNA for bikunin and heavy chain 3 (H3) of IalphaI. Neither IL-1beta (1 ng/mL) nor 25 mmol/L D-glucose influenced their mRNA expression nor protein synthesis. In contrast, the addition of either IL-1beta or 25 mmol/L D-glucose increased TSG-6 mRNA expression. This was accompanied by an early up-regulation of TSG-6 protein expression following IL-1beta stimulation (24 h) and a late up regulation after the addition of 25 mmol/L D-glucose (96 h) in the cell culture supernatant and associated with the cell membranes. Early induction of TSG-6 mRNA by IL-1beta was unaffected by the addition of the protein synthesis inhibitor cycloheximide. In contrast, the later glucose-stimulated induction of TSG-6 mRNA was abrogated by the addition of cycloheximide. Stimulation of TSG-6 by either IL 1beta or 25 mmol/L D-glucose was associated with an inhibition of total percentage plasmin activity. Immunoprecipitation of TSG-6 in these samples returned plasmin activity to control levels. CONCLUSIONS: : The data demonstrate that human PTCs constitutively express the bikunin and H3 components of the IalphaI family of serum protease inhibitors. Moreover, the addition of IL-1beta or 25 mmol/L D-glucose up-regulates the expression of TSG-6 in these cells, resulting in an inhibition of plasmin activity. PMID- 11422745 TI - Enhanced expression of complement C5a receptor mRNA in human diseased kidney assessed by in situ hybridization. AB - BACKGROUND: Anaphylatoxin C5a mediates inflammatory responses through interaction with a specific C5a receptor (C5aR), the expression of which is thought to be restricted to peripheral blood leukocytes. Although the presence of C5aR on cultured mesangial cells and tubular epithelial cells has recently been documented, the tissue distribution of C5aR in diseased kidney has not yet been determined. METHODS: Immunohistochemistry and nonradioactive in situ hybridization for C5aR were performed in 34 tissue samples of kidneys from patients with various renal diseases, including 4 with minimal change nephrotic syndrome (MCNS), 5 with membranous nephropathy (MN), and 25 with mesangial proliferative glomerulonephritis (mesGN; 15 patients with IgA nephropathy, 5 with non-IgA mesGN, and 5 with lupus nephritis). Normal portions of surgically resected kidney served as the control. RESULTS: In normal kidneys, C5aR protein was detected in tubular epithelial cells, while C5aR mRNA was detected in a few glomerular cells, tubular epithelial cells, and vascular endothelial and smooth muscle cells. In MCNS, the distribution of C5aR protein and mRNA was similar to that in normal kidneys. In MN and mesGN, C5aR protein and mRNA were detected in mesangial cells, glomerular epithelial and endothelial cells, Bowman's capsule cells, tubular cells, infiltrating cells, and vascular endothelial and smooth muscle cells. The glomerular expression of C5aR mRNA and protein correlated positively with the degree of mesangial hypercellularity and mesangial matrix expansion in mesGN. In the tubulointerstitium, interstitial expression of C5aR mRNA correlated positively with the degree of tubular atrophy and interstitial broadening in mesGN. Furthermore, the interstitial expression of C5aR mRNA correlated positively with the level of serum creatinine. CONCLUSIONS: Our results indicate that renal cells produce C5aR and that activation of C5a/C5aR pathway on renal cells may be involved in tissue injury in mesGN. PMID- 11422746 TI - Fibroblast growth factor receptors and their ligands in the adult rat kidney. AB - BACKGROUND: Fibroblast growth factors (FGFs) are a family of at least 21 heparin binding proteins involved in many biological processes, both during development and in the adult, including cell proliferation, differentiation, and angiogenesis. FGFs mediate their effects through high-affinity tyrosine kinase receptors (FGFRs), which are encoded by four genes. The aims of the present study were to localize FGFR-1 through FGFR-3 in the normal adult rat kidney and to determine which functional FGFR variants and FGFs were expressed. METHODS: Avidin biotin-enhanced horseradish peroxidase immunohistochemistry was used on paraffin sections of rat kidney to localize FGFR-1 through FGFR-3, whereas reverse transcriptase-polymerase chain reaction was used to examine expression of the receptor variants and also of FGF-1 through FGF-10 in cortex, outer medulla, and inner medulla. RESULTS: By immunohistochemistry, each receptor was localized to distinct and overlapping nephron segments, such that one or more FGFRs were localized to all nephron and collecting duct epithelia. FGFR-1 and FGFR-3 were localized to glomeruli, FGFR-3 to proximal tubules and FGFR-1 to thin limbs. FGFR 1 through FGFR-3 were localized to distal straight tubules, with FGFR-1 and FGFR 3 localized to distal convoluted tubules. FGFR-1 and FGFR-3 were localized to medullary collecting ducts. In addition, FGFR-1 was localized to the smooth muscle of renal arteries. All seven FGFR variants were expressed in the cortex and outer medulla, with fewer FGFRs in the inner medulla. FGF-1, FGF-2, FGF-7, FGF-8, and FGF-9 were expressed in the kidney, with FGF-10 expression found only in the cortex. CONCLUSIONS: Mapping of these receptors is critical to the determination of the effects of FGF ligands in discrete regions of the kidney. The distributions of the FGFRs in the normal adult kidney and the restricted expression of FGF ligands suggest that specific FGFs have distinct and important roles in the maintenance of normal kidney structure and function. PMID- 11422747 TI - Expression of multidrug resistance P-glycoprotein in kidney allografts from cyclosporine A-treated patients. AB - BACKGROUND: The multidrug resistance (MDR) gene product P-glycoprotein (P-gp) is a transmembrane efflux pump for hydrophobic, potentially toxic compounds, including the immunosuppressant cyclosporine A (CsA). We have previously shown that CsA increases P-gp expression in proximal tubule and endothelial cells in vitro. The aim of the present study was to investigate the in vivo relevance of these observations in renal allograft biopsies from CsA-treated patients. METHODS: P-gp expression was determined by immunohistochemistry of paraffin sections using two different monoclonal antibodies (UIC2 and MRK16). Biopsies were taken from CsA-treated renal transplant patients with different histopathological diagnoses (N = 79) and were compared with biopsies from normal human kidneys (N = 13) or with allograft biopsies from patients under a CsA-free immunosuppression (N = 15). Moreover, biopsies from 10 donor kidneys before implantation and during rejection episodes ("zero biopsies") were investigated. RESULTS: P-gp expression in biopsies with acute tubular necrosis (ATN; N = 10) after CsA treatment was significantly higher in arterial endothelia, proximal tubules, and epithelial cells of Bowman's capsule (BC), whereas P-gp was sparsely induced in CsA nephrotoxicity (N = 19) compared with controls. Acute cellular (N = 30) and vascular rejection (N = 10) or chronic allograft nephropathy (N = 10) after CsA was associated with strong P-gp expression in infiltrating leukocytes and increased P-gp expression in arterial endothelia, proximal tubules, and BC. In contrast, biopsies of patients treated with a CsA-free immunosuppression regimen did not show increases in P-gp expression compared with controls. Zero biopsies showed a weak, homogeneous, nonpolarized expression of P-gp in tubules and an increased expression of P-gp after CsA therapy in the brush border, arterial endothelia, and BC. CONCLUSIONS: CsA treatment was associated with increased P-gp expression in parenchymal cells of kidney transplants with ATN, acute or chronic transplant rejection, but P-gp was not increased in patients with CsA nephrotoxicity. This indicates that CsA induces its own detoxification by P-gp and that inadequate up-regulation of P-gp in renal parenchymal cells contributes to CsA nephrotoxicity. Increased expression of P-gp in infiltrating leukocytes correlated with the severity of allograft rejection, suggesting that P gp may decrease the immunosuppressive efficacy of CsA. Thus, individual differences in the P-gp induction response of CsA-exposed renal parenchymal cells and/or infiltrating leukocytes may predispose to either CsA nephrotoxicity or rejection, respectively. PMID- 11422748 TI - Role of stem cell factor and mast cells in the progression of chronic glomerulonephritides. AB - BACKGROUND: Mast cells (MCs) have been implicated in the pathogenesis of atherosclerosis and tissue fibrosis. However, the role of MC in the development of renal fibrosis has not been fully elucidated. Stem cell factor (SCF; the ligand for MC c-kit receptor) is thought to attract and activate MCs. METHODS: The intensity of MC infiltration and SCF expression in renal biopsies from 56 patients with different forms of primary and secondary glomerulonephritis and five controls were investigated by immunohistochemistry, using a monoclonal anti human MC tryptase antibody and a polyclonal antihuman SCF antibody. RESULTS: A large number of MCs were detected in the renal interstitium of the diseased kidneys. Immunostainable SCF was detected in tubular as well as interstitial cells. MC infiltration was significantly higher in glomerulonephritis (16.9 +/- 10.2 cells/field) compared with controls (2.8 +/- 2.1 cells/field, P = 0.03). Similarly, immunostainable SCF was 0.6 +/- 0.3% for controls and 3.3 +/- 2.1% in the glomerulonephritis group (P = 0.02). MC infiltration was highly correlated with SCF expression in diseased kidneys (r = 0.93, P = 0.0001). Double immunostain showed them to colocalize in some interstitial cells. Analysis of MC proliferation [proliferating cell nuclear antigen (PCNA) positivity] and apoptosis (in situ end labeling of DNA) showed these cells to be terminally differentiated. Both MCs and SCF were correlated with interstitial fibrosis (R = 0.71 for MC and R = 0.62 for SCF, P = 0.0001) and interstitial alpha-smooth muscle actin (R = 0.69 for MC and R = 0.60 for SCF P = 0.0001). Using regression analysis, the number of MC infiltration was found to be a very powerful determinant of interstitial fibrosis in the glomerulonephritis group (R2 = 91.4%). CONCLUSION: MCs as an infiltrating hematopoietic cell and its growth factor (SCF) seem to be up-regulated in glomerulonephritis, and may play a role in the development of renal fibrosis. PMID- 11422750 TI - Vascular function of the peripheral circulation in patients with nephrosis. AB - BACKGROUND: Nephrotic syndrome is associated with abnormal lipoprotein metabolism and increased risk of coronary heart disease. Endothelial dysfunction, an early phase of atherogenesis that manifests as impaired flow-mediated dilation (FMD) of the peripheral circulation, may link these associations. METHODS: We examined endothelial function of the brachial artery and forearm resistance arteries in 15 patients with nephrosis (NP), 15 patients with primary hyperlipidemia (HL) alone, and 15 normolipidemic, nonproteinuric subjects (NC) matched for age, sex, and weight. The NP and HL groups had similar serum cholesterol and triglyceride concentrations. Post-ischemic FMD (endothelium-dependent) and glyceryl trinitrate mediated dilation (GTNMD; endothelium-independent) of the brachial artery were studied using ultrasonography and computerized edge detection software. Postischemic forearm blood flow was also measured using plethysmography. RESULTS: Postischemic FMD of the brachial artery was significantly lower in the NP and HL groups compared with NC group (mean +/- SE): NP 4.91 +/- 0.8%, HL 4.53 +/- 0.6%, NC 8.45 +/- 0.5% (P < 0.001). There were no significant differences among the groups in baseline diameter and GTNMD of the brachial artery, nor in maximal forearm blood flow and flow debt repayment of the forearm microcirculation. Significant differences in FMD among the groups were principally related to differences in serum low-density lipoprotein cholesterol. CONCLUSIONS: Patients with NP have abnormal endothelium-dependent but preserved endothelium-independent dilation of the brachial artery following an ischemic stimulus. Postischemic forearm microcirculatory function is unimpaired. Dyslipoproteinemia is probably the principal cause of endothelial dysfunction of conduit arteries in patients with NP and the basis for their increased risk of cardiovascular disease. PMID- 11422749 TI - Role of glucocorticoids in the maturation of the rat renal Na+/H+ antiporter (NHE3). AB - BACKGROUND: Neonates have a lower Na+/H+ antiporter activity on the apical membrane of proximal tubule than that of adults. The maturational increase in Na+/H+ antiporter activity occurs at the time when there is a rise in serum glucocorticoid levels in rats. The purpose of the present study was to examine whether glucocorticoids are responsible for the postnatal increase in Na+/H+ antiporter activity. METHODS: Nine-day-old Sprague-Dawley rats were compared with rats studied at 30 days of age who had either a sham operation or adrenalectomy (ADX) at nine days of age and with rats that had an adrenalectomy and physiologic corticosterone replacement (ADX-Cort) to determine whether glucocorticoid deficiency prevented the maturational increase in Na+/H+ antiporter activity. Na+/H+ antiporter activity was measured in proximal convoluted tubules perfused in vitro by the change in cell pH (pHi) following luminal sodium removal. NHE3 mRNA abundance was measured using Northern blot analysis, and NHE3 protein abundance was measured by immunoblot. RESULTS: Na+/H+ antiporter activity was 93.8 +/- 17.7, 157.0 +/- 18.0, 356.7 +/- 29.9, and 402.5 +/- 14.5 pmol/mm. min in nine-day-old, ADX, ADX-Cort, and sham control groups, respectively. The ADX-Cort and sham control were higher than the 9-day-old and the 30-day-old ADX group (P < 0.05). Brush-border membrane NHE3 protein abundance in the nine-day-old and ADX groups were sixfold less than ADX-Cort and sham control groups (P < 0.001). Nine day-old neonates had fivefold less renal cortical NHE3 mRNA than the ADX, ADX Cort, and sham-operated control groups (P < 0.01). CONCLUSIONS: These data demonstrate that glucocorticoids play a role in the postnatal maturation of the proximal tubule Na+/H+ antiporter activity and brush-border membrane NHE3 protein abundance. Glucocorticoid deficiency does not completely prevent the maturational increase in Na+/H+ antiporter activity and does not affect NHE3 mRNA abundance. PMID- 11422751 TI - Connexins 40 and 43 are differentially regulated within the kidneys of rats with renovascular hypertension. AB - BACKGROUND: Connexin 40 (Cx40) is a gap junction protein expressed in endothelial cells of vessels, endocardium, and conducting cells of the His-Purkinje system of heart. Its distribution in the kidney remains to be fully investigated. METHODS: To address this distribution, we generated rabbit antibodies directed to a polypeptide comprising amino acids 231 to 330 of the carboxy-terminal domain of rodent Cx40, which specifically recognizes this protein at gap junctions. RESULTS: Immunolabeling and in situ hybridization of kidney sections showed that Cx40 and its transcript were coexpressed by most endothelial cells of vessels and glomeruli, as well as by renin-secreting cells. This distribution contrasted with that of Cx43, which was expressed in some tubules of medulla and sparse endothelial cells. Cx40 and Cx43 expression were investigated further in a renin dependent model of hypertension, in which rats showed an increase in arterial mean blood pressure four weeks after clipping one renal artery [two kidney, one clip (2K1C) model]. Northern blot analysis of polyA+ RNA demonstrated that, compared with sham-operated animals, the hypertensive 2K1C animals featured an increase in Cx40 mRNA expression in both left (clipped) and right (unclipped) kidneys. In contrast, Cx43 mRNA expression was only increased in the latter organ. Antibodies confirmed that the levels of Cx40 were actually increased in the kidneys of hypertensive animals (Western blots) and this was caused, at least in part, by enhanced expression of this protein in the renin-secreting cells of the afferent arteriole (immunofluorescence). CONCLUSIONS: Cell-to-cell communication mediated by Cx40 may be implicated in the function of renin secreting cells, hence participating in the control of blood pressure. PMID- 11422752 TI - Diabetes-induced microvascular dysfunction in the hydronephrotic kidney: role of nitric oxide. AB - BACKGROUND: Renal hemodynamics in early diabetes are characterized by preglomerular and postglomerular vasodilation and increased glomerular capillary pressure, leading to hyperfiltration. Despite intensive research, the etiology of the renal vasodilation in diabetes remains a matter of debate. The present study investigated the controversial role of nitric oxide (NO) in the renal vasodilation in streptozotocin-induced diabetic rats. METHODS: In the renal microcirculation, basal tone and response to NO synthase blockade were studied using the in vivo hydronephrotic kidney technique. L-arginine analog N-nitro-L arginine methyl ester (L-NAME) was administered locally to avoid confounding by systemic blood pressure effects. The expression of endothelial NO synthase (eNOS) was investigated in total kidney by immunocytochemistry and in isolated renal vascular trees by Western blotting. Urinary excretion of nitrites/nitrates was measured. RESULTS: Diabetic rats demonstrated a significant basal vasodilation of all preglomerular and postglomerular vessels versus control rats. Vasoconstriction to L-NAME was significantly increased in diabetic vessels. After high-dose L-NAME, there was no difference in diameter between diabetic and control vessels, suggesting that the basal vasodilation is mediated by NO. Immunocytochemically, the expression of eNOS was mainly localized in the endothelium of preglomerular and postglomerular vessels and glomerular capillaries, and was increased in the diabetic kidneys. Immunoblots on isolated renal vascular trees revealed an up-regulation of eNOS protein expression in diabetic animals. The urinary excretion of nitrites/nitrates was elevated in diabetic rats. CONCLUSION: The present study suggests that an up-regulation of eNOS in the renal microvasculature, resulting in an increased basal generation of NO, is responsible for the intrarenal vasodilation characteristic of early diabetes. PMID- 11422753 TI - Aldose reductase expression is induced by hyperglycemia in diabetic nephropathy. AB - BACKGROUND: Despite good metabolic control, many patients with type 1 diabetes still develop nephropathy, implicating a role for genetic factors. Recent studies examining the regulatory region of the aldose reductase (ALR2) gene, the rate limiting enzyme of the polyol pathway, support its role as a candidate gene for nephropathy. Here we report the quantitation of ALR2, together with sorbitol dehydrogenase mRNA in the peripheral blood mononuclear cells (PBMCs) of type 1 diabetic patients with (N = 29) and without nephropathy (N = 11) following stimulation with high levels of D-glucose. METHODS: PBMCs from patients and normal controls were cultured for five days with phytohemagglutinin in either normoglycemia (11 mmol/L D-glucose) or supplemented with 10 mmol/L D-glucose (moderate hyperglyemia) or 20 mmol/L D-glucose (hyperglycemia). The RNA was extracted and analyzed by ribonuclease protection assay. RESULTS: ALR2 mRNA levels were significantly elevated with increasing D-glucose concentration (normal to hyperglycemic) in those patients with nephropathy (P < 0.0001). In marked contrast, in those without nephropathy and in the normal healthy controls, there was no change in mRNA expression. Furthermore, those patients with nephropathy and the Z-2/X susceptibility genotype had the greatest increase in ALR2 mRNA compared with those with low-risk genotypes (P < 0.007). CONCLUSION: These results show that patients with nephropathy exhibit marked disturbances in the expression of the enzyme components of the polyol pathway. Ultimately this leads to tissue damage and ischemia. PMID- 11422754 TI - Microalbuminuria in type 1 diabetes: rates, risk factors and glycemic threshold. AB - BACKGROUND: The occurrence of microalbuminuria in type 1 diabetes is strongly predictive of renal and cardiovascular disease and is still likely to occur despite improvements in glycemic control. A better understanding of microalbuminuria is required to inform new interventions. We determined the incidence and risk factors for microalbuminuria [albumin excretion rate (AER) 20 to 200 microg/min] in the EURODIAB Prospective Complications Study. METHODS: This is a seven-year follow-up (between 1988 and 1991) of 1134 normoalbuminuric men and women (aged 15 to 60) with type 1 diabetes from 31 European centers. Risk factors and AER were measured centrally. RESULTS: The incidence of microalbuminuria was 12.6% over 7.3 years. Independent baseline risk factors were HbA1c (7.1 vs. 6.2%, P = 0.0001) and AER (9.6 vs. 7.8 microg/min, P = 0.0001) and, independent of these, fasting triglyceride (0.99 vs. 0.88 mmol/L, P = 0.01), low-density lipoprotein cholesterol (3.5 vs. 3.2 mmol/L, P = 0.02), body mass index (24.0 vs. 23.4 kg/m2, P = 0.01), and waist to hip ratio (WHR; 0.85 vs. 0.83, P = 0.009). Triglyceride and WHR risk factors were nearly as strong as AER in predicting microalbuminuria (standardized regression effects of 1.3 for triglyceride and WHR and 1.5 for AER). Blood pressure at follow-up, but not at baseline, was also raised in those who progressed. There was no evidence of a threshold of HbA1c on microalbuminuria risk. CONCLUSIONS: The incidence of microalbuminuria in patients with type 1 diabetes remains high, and there is no apparent glycemic threshold for it. Markers of insulin resistance, such as triglyceride and WHR, are strong risk factors. Systemic blood pressure is not raised prior to the onset of microalbuminuria. PMID- 11422755 TI - Long-term beneficial effect of ACE inhibition on diabetic nephropathy in normotensive type 1 diabetic patients. AB - BACKGROUND: The purpose of this study was to assess whether long-term (8 years) inhibition of angiotensin-converting enzyme (ACE) protects kidney function in normotensive type 1 diabetic patients with diabetic nephropathy. METHODS: We performed an open randomized follow-up study of normotensive type 1 diabetics with nephropathy either treated (N = 15) or not (N = 17) with captopril twice per day (average 74, range 12.5 to 125 mg/day). The main outcome measures were arterial blood pressure, albuminuria, and glomerular filtration rate (GFR; 51Cr EDTA plasma clearance, twice yearly). RESULTS: Arterial blood pressure (mm Hg) was kept constant in the captopril group, at baseline (mean, SEM), 128/78 (3/2) and during follow-up 129/77 (4/1) but increased significantly in the control group from 127/79 (2/1) to 137/84 (5/2) (P < 0.01). Furthermore, 8 out of the 17 control subjects required treatment with blood pressure-lowering drugs because they developed hypertension. The fractional albumin clearance (x10-5) remained unchanged in the captopril group: baseline [10.8 (1.25) geometric mean and antilog (SEM)] during the eight years [11.8 (1.47)], while a significant rise occurred in control patients: 13.3 (1.23) to 26.2 (1.42) (P < 0.05). Baseline GFR was nearly identical: 111 (6) and 115 (4) mL/min/1.73 m2 in the captopril and control group, respectively. The median (range) rate of decline in GFR (mL/min/year) was 1.7 (10.7 to -2.0) in the captopril group versus 2.8 (17.7 to 2.6) in the control group (P = NS). CONCLUSIONS: The beneficial effect of captopril in arresting the rise in systemic blood pressure and albuminuria is long lasting. A loss in GFR is minimal in most patients with diabetic nephropathy if normotension is sustained by prospective treatment with ACE inhibitors or restored by implementation of other antihypertensive medications with the development of hypertension. PMID- 11422756 TI - Effects of dietary protein restriction on fibrinogen and albumin metabolism in nephrotic patients. AB - BACKGROUND: Nephrotic syndrome (NS) is characterized by profound changes in albumin and fibrinogen levels. Dietary protein restriction has been advocated in the treatment of patients with NS, but its effects on albumin and fibrinogen metabolism have not been fully elucidated. METHODS: We evaluated the effects of dietary protein restriction on endogenous leucine flux (ELF), fibrinogen and albumin metabolism in seven patients with NS who consumed either a normal protein diet (NPD; 1.20 +/- 0.06 g/kg/day), or a low protein diet (LPS; 0.66 +/- 0.04 g/kg/day) for four weeks. Seven normal subjects served as controls. The postabsorptive ELF value, fractional synthesis rate (FSR) and absolute synthesis rate (ASR) of both albumin and fibrinogen were evaluated during the last 120 minutes of a five-hour 5,5,5-D3-L-leucine infusion. RESULTS: During the NPD regimen. ELF was increased, serum albumin was reduced, plasma fibrinogen was increased, albumin FSR and ASR were both increased, fibrinogen FSR was normal, and fibrinogen ASR was greater in patients with NS compared to controls. In patients with NS the LPD regimen reduced proteinuria, ELF, albumin FSR and ASR, plasma fibrinogen levels, fibrinogen ASR, and increased serum ulbumin levels. Dietary-induced changes in albumin and fibrinogen synthesis were significantly correlated (r = 0.719, P < 0.05). CONCLUSIONS: Patients with NS treated with LPD show: (1) a reduction of proteinuria, albumin ASR and FSR, with an increase in serum albumin levels and its intravascular pool; (2) a decrease of fibrinogen ASR, with a reduction in both plasma fibrinogen levels and intravascular pool; and (3) a reduced rate of whole body proteolysis. PMID- 11422757 TI - The natural history of renal disease in Australian Aborigines. Part 1. Changes in albuminuria and glomerular filtration rate over time. AB - BACKGROUND: The purpose of this study was to describe changes over time in albuminuria and glomerular filtration rate (GFR) in a cohort of Australian Aborigines from a community with high rates of renal disease and renal failure. METHODS: Participants were 486 adult community members (20+ years at first exam) who were screened for renal disease and related factors on at least two occasions (mean 2.7 occasions), at least a year apart, between 1990 and 1997. Renal function was assessed by the albumin:creatinine ratio (ACR; g/mol) on a random urine specimen and by the GFR estimated from the Cockcroft-Gault formula. Evolution over time was expressed as the average annual changes in these parameters. RESULTS: On baseline examination, 70% of participants had albuminuria (ACR 1.1+ g/mol) There was a significant net increase in ACR and a fall in GFR in the cohort over time. Among individuals, however, changes were strongly correlated with ACR levels at baseline. There was no loss of GFR in persons with normal renal parameters at baseline and a rapid loss of GFR in those with substantial levels of albuminuria at baseline. Other factors significantly correlated with progression of ACR included age, baseline body mass index and systolic blood pressure, the presence of diabetes (or levels of fasting glucose), and elevated levels of serum gamma glutamyl transferase. Factors significantly associated with loss of GFR included body mass index, diabetes, systolic and diastolic blood pressures, microscopic hematuria, and marginally high cholesterol levels. CONCLUSION: Albuminuria progresses and GFR is lost over time in individuals in this community, at rates that are strongly dependent on levels of pre-existing albuminuria. Much loss of GFR and all renal failure should be avoided by preventing the development of albuminuria and minimizing its progression. This depends on improving the weight, blood pressure, and metabolic profile of the entire community and reducing infections. Modification of the course in people with established disease depends on vigorous control of blood pressure and the metabolic profile and the specific use of angiotensin-converting enzyme inhibitors. PMID- 11422758 TI - The natural history of renal disease in Australian Aborigines. Part 2. Albuminuria predicts natural death and renal failure. AB - BACKGROUND: The purpose of this study was to describe the relationship of albuminuria and glomerular filtration rate (GFR) with natural death and renal failure in an Australian Aboriginal community with high rates of renal disease. METHODS: Study subjects were 825 adults (18+ years, mean 33.6 years) or 88% of adults in a remote community who participated in a health screening program offered between 1990 and 1997. The urinary albumin:creatinine ratio (ACR; g/mol) was used as the renal disease marker. Participants were followed for 1.0 to 9.8 years (mean 5.8 years) until renal failure, death, the start of systematic antihypertensive/renal-protective treatment or June 30, 2000. RESULTS: Sixty-five people reached a terminal end point of renal failure or natural death. Sixteen people developed terminal renal failure, all of whom had an ACR of 34+ at baseline exam. There were 49 other natural deaths, which were also strongly correlated with increasing ACR and decreasing GFR over a wide range. This was observed in people without diabetes and in people with normal and elevated blood pressures. It applied to deaths associated with cardiovascular disease and to deaths without an assigned primary or underlying cardiovascular or renal cause. With adjustment for age, the association with death was more robust with ACR than GFR. When compared with people with an ACR <3.4, the hazard ratio (HR; 95% CI) for nonrenal natural death of persons with an ACR 3.4 to 33 was 3.0 (1.1 to 8.4), with an ACR 34 to 99, it was 5.4 (1.8 to 15.9), and with an ACR 100+, it was 6.5 (2.0 to 21). Regression equations predicted that each tenfold increase in the ACR was associated with a 3.7-fold increase in all-cause natural death: a> 400-fold increase in renal deaths, a 4-fold increase in cardiovascular deaths, and a 2.2 fold increase in nonrenal noncardiovascular deaths. Eighty-four percent of all cause natural death was associated with pathologic albuminuria. CONCLUSION: All renal failure develops out of a background of persistent albuminuria in this population. More important, albuminuria and, inversely, GFR are powerful markers of risk for nonrenal natural death, including, but not restricted to, cardiovascular deaths. Most of the risk for premature death can be assessed by a simple urine test, and interventions that prevent development and progression of albuminuria and loss of GFR should not only prevent renal insufficiency, but powerfully reduce mortality from natural causes as well. PMID- 11422759 TI - Effect of chronic renal failure on bone turnover and bone alkaline phosphatase isoforms. AB - BACKGROUND: Biochemical markers of bone turnover are used to monitor metabolic bone disease associated with renal failure. We have applied a comprehensive panel of markers to patients with chronic renal failure (CRF), with particular focus on the isoforms of bone alkaline phosphatase (BALP). METHODS: Twenty CRF patients undergoing hemodialysis (N = 9) and peritoneal dialysis (N = 11) were measured for serum parathyroid hormone (PTH), osteocalcin, total ALP, and four BALP isoforms (B/I, B1x, B1, and B2) by high-performance liquid chromatography. These BALP isoforms were also compared with BALP measured by three commercial immunoassays (Alkphase-B, Tandem-R Ostase, and Tandem-MP Ostase). Type I collagen turnover was assessed by serum samples using the type I procollagen intact amino- and carboxy-terminal propeptides (PINP and PICP) and two fragments (ICTP and CrossLaps) derived from the carboxy-terminal telopeptide of mature matrix collagen by different degradative pathways. RESULTS: Mean levels of bone turnover markers were elevated in CRF, with marked increases in those markers, osteocalcin, ICTP, and CrossLaps, cleared by the kidney. Total ALP activities were increased corresponding to elevated B/I and B2 isoform levels. The B1 isoform level was not significantly different from healthy controls. B1x was detected in 60% of the patients but was not resolved in healthy individuals. Kendall's tau rank correlation showed that B1x correlated significantly (P < 0.05) with B1 (0.53) and PINP (0.55), and was the only marker to correlate with PTH (0.49). B1x was not significantly correlated with any of the commercial BALP immunoassays. Interestingly, the immunoassay calibrators contained high activities of the B/I peak (39 to 80%) compared with human serum (4%). CONCLUSION: There are selective differences between the BALP isoforms in CRF compared with healthy adults. The commercial BALP immunoassays are comparable with each other but are unable to distinguish the BALP isoform-specific differences in CRF patients. PMID- 11422760 TI - Lead chelation therapy and urate excretion in patients with chronic renal diseases and gout. AB - BACKGROUND: It is known that chronic renal insufficiency (CRI) patients with gout may have subtle lead poisoning. In addition, gout episodes frequently aggravate progressive renal insufficiency because of the use of nephrotoxic drugs and urate deposition. Our study was arranged to evaluate the causal effect of environmental lead exposure on urate excretion in CRI patients. METHODS: A cross-section study and a randomized, controlled trial were performed. Initially, 101 patients with CRI and without a history of previous lead exposure received ethylenediaminetetraacetic acid mobilization tests to assess body lead stores (BLS). Then, a clinical trial was performed; 30 CRI patients with gout and high normal BLS and the changes of urate excretion in these patients were compared before and after lead chelating therapy. The treated group received four-week chelating therapy, and the control group received a placebo therapy. RESULTS: The BLS of patients with CRI and gout was higher than that of patients with CRI only, and none had subtle lead poisoning. The BLS, not the blood lead level (BLL), significantly correlated to indices of urate excretion in all CRI patients after related factors were adjusted. In addition, after lead chelating therapy, urate clearance markedly improved after a reduction of the BLS of patients with CRI and gout (study group 67.9 +/- 80.0% vs. control group 1.2 +/- 34.0%, P = 0.0056). CONCLUSION: Our findings suggest that the chronic low-level environmental lead exposure may interfere with urate excretion of CRI patients. Importantly, the inhibition of urate excretion can be markedly improved by lead chelating therapies. These data shed light on additional treatment of CRI patients with gout; however, more studies are needed to confirm our findings. PMID- 11422761 TI - Frequency of renal phosphate leak among patients with calcium nephrolithiasis. AB - BACKGROUND: Nephrolithiasis is a frequent disorder affecting 10 to 15% of the population in Europe and the United States. More than 80% of renal stones are made of calcium oxalate and calcium phosphate. The main identified risks for calcium renal stone formation are hypercalciuria and urinary saturation. A urine phosphate (Pi) loss is often associated with hypercalciuria; furthermore, hyperphosphaturia increases urinary saturation. METHODS: To determine whether urinary phosphate loss is associated with calcium urolithiasis, we measured renal Pi threshold (TmPi) in 207 stone formers with normal parathyroid hormone (PTH) serum concentration and in 105 control subjects. RESULTS: The TmPi followed a normal distribution in both groups. The mean TmPi was significantly lower in stone formers versus controls (0.72 +/- 0.13 vs. 0.87 +/- 0.18 mmol/L, P < 0.0001) because of a shift to the left of the TmPi distribution curve in the stone former population, with no evidence for bimodal distribution. Five percent of the controls had a TmPi <0.63 versus 19% of the stone formers. Daily urinary calcium excretion was significantly higher in stone formers than in controls. Calcium excretion was also significantly higher in stone formers with TmPi <0.63 mmol/L compared with those with TmPi > or =0.63. Serum PTH and ionized calcium concentrations were not different in stone formers and in control subjects, whatever the TmPi value. CONCLUSIONS: : A low TmPi is more frequently encountered in stone formers with a normal PTH concentration than in control subjects and is associated with a high urinary Ca excretion. The hypophosphatemia induced by a renal phosphate leak may predispose the subject to calcium stone formation by increasing the serum calcitriol level, calcium excretion, and urinary saturation. PMID- 11422762 TI - Remission and regression in the nephropathy of type 1 diabetes when blood pressure is controlled aggressively. AB - BACKGROUND: Diabetic nephropathy is a chronic, progressive kidney disease with a mean rate of decline of in glomerular filtration rate (GFR) of 10 to 12 mL/min/year (natural history). The introduction of aggressive antihypertensive treatment has improved the renal prognosis during the last decades. To examine whether remission and regression of diabetic nephropathy are possible in type 1 diabetic patients, we analyzed data from a prospective observational cohort study that was started in 1983. METHODS: We measured GFR with a 51Cr-EDTA plasma clearance technique every year for seven years (range 3 to 14 years) in 301 consecutive type 1 diabetic patients with diabetic nephropathy. Diabetic nephropathy was diagnosed clinically if the following criteria were fulfilled: persistent albuminuria> 200 microg/min, presence of diabetic retinopathy, and no evidence of other kidney or renal tract disease. Blood pressure, albuminuria, glycosylated hemoglobin A1c, and serum cholesterol were measured every three to four months during the study. In total, 271 patients received antihypertensive treatment, 179 patients predominantly with angiotensin-converting enzyme inhibitors. Remission was defined as albuminuria <200 microg/min sustained for at least one year and a decrease of at least 30% from preremission levels (surrogate endpoint), and regression as a rate of decline in GFR (DeltaGFR) equal to the natural aging process: < or =1 mL/min/year during the entire observation period (principal end point). RESULTS: The total number of patients who obtained remission was 92 (31%), with a duration of remission of [median (range)] 3.4 (1.0 to 14.1) years, and regression 67 (22%). The patients were stratified in quintiles by the average value of office mean arterial blood pressure (mean +/- SE): 93 +/- 0.5, 99 +/- 0.2, 103 +/- 0.1, 107 +/- 0.2, and 113 +/- 0.4 mm Hg. The prevalence of patients obtaining remission/regression was 58/42, 33/32, 25/11, 20/20, and 17/7% in each quintile, respectively. Spontaneous remission and regression occurred in 10 and 14 patients from the persistent normotensive group (N = 30), none of whom had ever received antihypertensive treatment. In all 301 consecutive patients, the (mean +/- SE) DeltaGFR was 4.0 +/- 0.2 mL/min/year during the investigation period. CONCLUSIONS: Our study suggests that aggressive antihypertensive treatment in type 1 diabetic patients can induce remission and regression in a sizable fraction of patients with diabetic nephropathy. Lower arterial blood pressure, reduced albuminuria, and better glycemic control were predictors of regression of diabetic nephropathy. PMID- 11422763 TI - Noninvasive transcutaneous determination of access blood flow rate. AB - BACKGROUND: Current indicator dilution techniques for determining the vascular access blood flow rate (Qa) require reversal of the dialysis blood lines and are time consuming. We have recently described an indicator dilution technique for determining Qa using a novel optical transcutaneous hematocrit (Hct) sensor that does not require reversal of the dialysis lines, and have validated the accuracy of this method (TQa) in vitro. METHODS: This study compared results using the TQa method with those obtained using a similar indicator dilution technique but which required reversal of the dialysis lines (HD01 Monitor, Transonic Systems, Ithaca, NY, USA) during routine hemodialysis in 59 patients (25 native fistulas and 34 synthetic grafts). The sensor for the TQa method was placed on the skin directly over the access to measure changes in Hct approximately 25 mm downstream of the venous needle. A single 30 mL bolus of saline was infused into the dialyzer venous line over approximately six seconds without reversal of the dialysis blood lines, and the vascular access flow rate was calculated using indicator dilution methods from the time-dependent decrease in the Hct downstream of the venous needle. Two additional small-scale studies were performed to assess the effect skin pigmentation and to evaluate further the reproducibility of the TQa method. RESULTS: Qa values determined by the TQa method were highly correlated with those determined by the HD01 method (N = 72, R2 = 0.948, P < 0.001) over the range of 153 to 2,042 mL/min. There was no significant difference between vascular access flow rates determined by the TQa method and those determined by the HD01 METHOD: Results from one small-scale study showed that the relationship between Qa values determined by the TQa and the HD01 methods was similar when tested only among black patients (N = 12), suggesting that skin pigmentation is not an important determinant of the accuracy of the TQa METHOD: The second small-scale study showed that the intratreatment coefficient of variation for the TQa method was 7.8 +/- 5.6% (N = 14). CONCLUSIONS: : These results show that transcutaneous measurement of Qa is an accurate, simple, and fast technique for determining Qa without requiring the reversal of the dialysis blood lines. PMID- 11422764 TI - Differential survival after coronary revascularization procedures among patients with renal insufficiency. AB - BACKGROUND: Acute myocardial infarction, cardiac arrest, and other cardiac events are the major cause of mortality among patients with renal insufficiency. Previous studies of interventions for coronary artery disease among patients with renal insufficiency have not controlled for potentially confounding factors such as coronary artery disease severity and left ventricular function. This study investigates the comparative survival for patients with renal insufficiency and coronary artery disease following coronary artery bypass graft (CABG) surgery as compared with percutaneous coronary artery intervention (PCI), while controlling for confounding factors. METHODS: This retrospective cohort study of patients undergoing CABG surgery or PCI discharged between 1993 and 1995 uses the New York Department of Health databases and Cox proportional hazards analyses to estimate the mortality risk associated with CABG as compared with PCI for patients with renal insufficiency. Renal function was categorized as creatinine <2.5 mg/dL (N = 58,329), creatinine > or =2.5 mg/dL (N = 840), and end-stage renal disease (ESRD) requiring dialysis (N = 407). RESULTS: Patients with either ESRD or serum creatinine > or =2.5 mg/dL had more severe coronary artery disease and a greater frequency of comorbid conditions as compared with patients with creatinine <2.5 mg/dL. Creatinine > or =2.5 mg/dL and ESRD were both associated with an increased mortality risk among all distributions of coronary artery disease anatomy. Among patients with ESRD, the risk ratio (RR) of mortality for patients undergoing CABG compared with PCI was 0.39 (95% CI, 0.22 to 0.67, P = 0.0006). Among patients with creatinine > or =2.5 mg/dL, CABG surgery did not convey a survival benefit over PCI (RR, 0.86, 95% CI, 0.56 to 1.33, P = 0.50). CONCLUSIONS: This study demonstrates a survival benefit among patients with ESRD undergoing CABG surgery as compared with PCI, while controlling for severity of coronary artery disease, left ventricular dysfunction, and other comorbid conditions. These results suggest that management decisions among patients with coronary artery disease should be made in the context of not only location and severity of coronary artery lesions, but also on the presence and severity of renal dysfunction. PMID- 11422765 TI - Diagnostic value of iron indices in hemodialysis patients receiving epoetin. AB - BACKGROUND: Iron deficiency remains a common cause of hyporesponsiveness to epoetin in hemodialysis patients. However, considerable controversy exists regarding the best strategies for diagnosis and treatment. METHODS: As part of a multicenter randomized clinical trial of intravenous versus subcutaneous administration of epoetin, we made monthly determinations of serum iron, total iron binding capacity, percentage transferrin saturation, and serum ferritin. If a patient had serum ferritin <100 ng/mL or the combination of serum ferritin <400 ng/mL and a transferrin saturation <20%, he/she received parenteral iron, given as iron dextran 100 mg at ten consecutive dialysis sessions. We analyzed parenteral iron use during the trial, the effect of its administration on iron indices and epoetin dose, and the ability of the iron indices to predict a reduction in epoetin dose in response to parenteral iron administration. RESULTS: Eighty-seven percent of the 208 patients required parenteral iron to maintain adequate iron stores at an average dose of 1516 mg over 41.7 weeks, or 36 mg/week. Only two of 180 patients experienced serious reactions to intravenous iron administration. Two thirds of the patients receiving parenteral iron had a decrease in their epoetin requirement of at least 30 U/kg/week compared with 29% of patients who did not receive iron (P = 0.004). The average dose decrease 12 weeks after initiating iron therapy was 1763 U/week. A serum ferritin <200 ng/mL had the best positive predictive value (76%) for predicting a response to parenteral iron administration, but it still had limited clinical utility. CONCLUSIONS: Iron deficiency commonly develops during epoetin therapy, and parenteral iron administration may result in a clinically significant reduction in epoetin dose. The use of transferrin saturation or serum ferritin as an indicator for parenteral iron administration has limited utility. PMID- 11422766 TI - Post-transplant renal artery stenosis: the hemodynamic response to revascularization. AB - BACKGROUND: Percutaneous transluminal angioplasty and stenting are relatively noninvasive approaches to treat post-transplant renal artery stenosis. However, the real impact of this procedure on renal function recovery has never been quantitated precisely to date. METHODS: In eight consecutive renal transplant patients with renal graft artery stenosis, blood pressure, body weight, and anatomical, functional, and Doppler ultrasound parameters were evaluated before and one month after renal artery transluminal angioplasty and stenting. On both occasions, glomerular filtration rate and renal plasma flow were evaluated by inulin and paraaminohippuric acid renal clearances, and glomerular size-selective function was evaluated by the fractional clearances of neutral dextran macromolecules. RESULTS: The correction of renal artery stenosis, by normalizing renal vascular resistances, fully restored kidney perfusion and decreased arterial blood pressure, relieved water and sodium retention, restored an almost laminar arterial blood flow, and normalized vascular shear stress without appreciable effects on glomerular barrier size-selective function and proteinuria. Preangioplasty and postangioplasty renal resistive indices and peak systolic blood velocity estimated by Doppler ultrasounds were significantly correlated with the effective renal plasma flow and the blood velocity calculated at the site of stenosis. All patients were discharged without sequelae one or two days after angioplasty. CONCLUSIONS: Percutaneous transluminal angioplasty and stenting are safe and effective procedures to normalize the functional changes sustained by hemodynamically significant artery stenosis after renal transplantation. Doppler ultrasound scanning is a reliable and reproducible technique to monitor the renal functional response to vascular reperfusion. PMID- 11422767 TI - Influence of various hemodialysis membranes on the plasma (1-->3)-beta-D-glucan level. AB - BACKGROUND: Recently, invasive fungal infections have increased significantly because of the growing number of immunocompromised hosts. The measurement of plasma (1-->3)-beta-D-glucan has been proposed as a useful diagnostic tool for deep mycosis. In this study, we analyzed the alteration of the plasma (1-->3) beta-D-glucan concentration by using different kinds of hemodialysis (HD) membranes in end-stage renal disease (ESRD) patients and estimated its half-life. METHODS: Twenty-seven patients with ESRD without known fungal infections (with a median age of 66 years old) were enrolled in this study. Cellulose triacetate (CTA) membrane, cellulose (Cu) membrane, and polymethyl methacrylate (PMMA) membrane were used in three consecutive initiations of HD and plasma (1-->3)-beta D-glucan concentration was measured before and after each HD session. RESULTS: The level did not change between before and after HD when CTA and PMMA membranes were used. In contrast, the plasma (1-->3)-beta-D-glucan level increased greatly after HD using the Cu membrane (from 9.4 to 332 pg/mL in median). Hypothesizing that the plasma (1-->3)-beta-D-glucan level declines exponentially, its median half-life is estimated at 20 hours, which is not affected by hepatic or renal function. CONCLUSIONS: Because the plasma (1-->3)-beta-D-glucan level increases dramatically after HD using the Cu membrane, dialyzers with Cu membrane should be avoided for HD in patients with suspected deep mycosis. The measurement of plasma (1-->3)-beta-D-glucan is also useful for the patients with hepatic or renal failure. PMID- 11422768 TI - Risk factors and mortality associated with calciphylaxis in end-stage renal disease. AB - BACKGROUND: We conducted a case control study to determine risk factors and mortality associated with calciphylaxis in end-stage renal disease. METHODS: Cases of calciphylaxis diagnosed between December 1989 and January 2000 were identified. Three controls were identified for each hemodialysis patient, with calciphylaxis matched to the date of initiation of hemodialysis. Laboratory data and medication doses were recorded during the 12 months prior to the date of diagnosis and at the time of diagnosis of calciphylaxis. Conditional logistic regression was used to identify risk factors for calciphylaxis. Cox proportional hazards models were used to estimate the risk of death associated with calciphylaxis. RESULTS: Nineteen cases and 54 controls were identified. Eighteen patients were hemodialysis patients, and one had a functioning renal allograft. Diagnosis was confirmed by skin biopsy in 16 cases. Women were at a sixfold higher risk of developing calciphylaxis (OR = 6.04, 95% CI 1.62 to 22.6, P = 0.007). There was a 21% lower risk of calciphylaxis associated with each 0.1 g/dL increase in the mean serum albumin during the year prior to diagnosis and at the time of diagnosis of calciphylaxis (OR = 0.79, 95% CI, 0.64 to 0.99, P = 0.037, and OR = 0.80, 95% CI, 0.67 to 0.96, P = 0.019, respectively). There was a 3.51 fold increase in the risk of calciphylaxis associated with each mg/dL increase in the mean serum phosphate during the year prior to diagnosis (95% CI, 0.99 to 12.5, P = 0.052). At the time of diagnosis of calciphylaxis, for each 10 IU/L increment in alkaline phosphatase, the risk of calciphylaxis increased by 19% (OR = 1.19, 95% CI, 1.00 to 1.40, P = 0.045). Body mass index, diabetes, blood pressure, aluminum, and higher dosage of erythropoietin and iron dextran were not independent predictors of calciphylaxis. Calciphylaxis independently increased the risk of death by eightfold (OR = 8.58, 95% CI, 3.26 to 22.6, P < 0.001). CONCLUSIONS: Female gender, hyperphosphatemia, high alkaline phosphatase, and low serum albumin are risk factors for calciphylaxis. Calciphylaxis is associated with a very high mortality. PMID- 11422769 TI - Inflammation and dietary protein intake exert competing effects on serum albumin and creatinine in hemodialysis patients. AB - BACKGROUND: Cross-sectional studies have shown an inverse correlation between serum C-reactive protein (CRP) and serum albumin concentration in hemodialysis patients. The net effects of inflammation and dietary protein intake on nutritional markers over time are unknown. METHODS: To explore the effects of CRP and normalized protein catabolic rate (nPCR) on serum albumin and creatinine, we analyzed six consecutive months of laboratory data from 364 hemodialysis patients, using a multivariable Mixed model with conservative biases. RESULTS: The overall trend over time in serum albumin was slightly positive (0.039 g/dL/month) and in serum creatinine slightly negative (-0.052 mg/dL/month). With increasing CRP, serum albumin declined significantly (-0.124 g/dL/month per unit increase in log CRP, adjusted for age, gender, race, diabetes, and nPCR, P < 0.0001). Serum albumin increased with increasing nPCR (0.021 g/dL/month per 0.1 g/kg/day, P < 0.0001). The effect of CRP on albumin was attenuated in African Americans and at a higher nPCR. Corresponding values for creatinine mirrored those for albumin. With increasing CRP, creatinine declined significantly [-0.142 mg/dL/month per unit increase in log CRP, adjusted for age, gender, race, diabetes (time since initiation of dialysis; vintage), Kt/V, and nPCR, P = 0.002]. Serum creatinine increased with increasing nPCR (0.183 mg/dL/month per g/kg/day, P < 0.0001). CONCLUSIONS: Proxies of inflammation and dietary protein intake exert competing effects on serum albumin and creatinine in hemodialysis patients. These data provide a rationale for prospective testing of dietary protein supplementation in hemodialysis patients with biochemical evidence of ongoing inflammation and "malnutrition." PMID- 11422770 TI - Multicenter trial of one HLA-DR-matched or mismatched blood transfusion prior to cadaveric renal transplantation. AB - BACKGROUND: The beneficial effect of blood transfusions before cadaveric renal transplantation on allograft survival, although previously well documented, has become controversial in light of their adverse effects. Recently, it has been suggested that their clinical benefits are due to HLA-DR sharing between the blood donor and recipient. METHODS: In this prospective study, 144 naive patients were randomly assigned to receive one unit of blood matched for one-HLA-DR antigen (N = 49), or one unit of mismatched blood (N = 48), or to remain untransfused (N = 47). Graft survival and acute rejection rate were analyzed in 106 cadaveric renal allograft recipients receiving the same immunosuppressive protocol. RESULTS: Graft survival was similar in the three groups at one and five years: 91.7 and 80% in untransfused patients, 90.3 and 79.3% in patients transfused with one DR-antigen-matched unit, and 92.3 and 83.7% in patients transfused with HLA-mismatched blood. The difference in the incidence of six month post-transplant acute rejections was not statistically significant in the three groups: 12 out of 36, 33.3% in nontransfused patients; 6 out of 31, 19.4% in patients transfused with one DR-matched blood; and 13 out of 39, 33.3% in patients transfused with mismatched blood. CONCLUSION: The results of our prospective randomized trial showed that in a population of naive patients, one transfusion mismatched or matched for one HLA-DR antigen given prior to renal transplantation had no significant effect on the incidence and severity of acute rejection, and did not influence overall long-term graft outcome. Considering the potentially deleterious adverse effects of blood transfusions, the costs, and the considerable logistical efforts required to select and type blood donors, such a procedure cannot be recommended in a routine practice for patients awaiting cadaveric kidney transplantation. PMID- 11422771 TI - Cardiac arrest and sudden death in dialysis units. AB - BACKGROUND: For patients with end-stage renal disease and their providers, dialysis unit-based cardiac arrest is the most feared complication of hemodialysis. However, relatively little is known regarding its frequency or epidemiology, or whether a fraction of these events could be prevented. METHODS: To explore clinical correlates of dialysis unit-based cardiac arrest, 400 reported arrests over a nine-month period from October 1998 through June 1999 were reviewed in detail. Clinical characteristics of patients who suffered cardiac arrest were compared with a nationally representative cohort of> 77,000 hemodialysis patients dialyzed at Fresenius Medical Care North America-affiliated facilities. RESULTS: The cardiac arrest rate was 400 out of 5,744,708, corresponding to a rate of 7 per 100,000 hemodialysis sessions. Cardiac arrest was more frequent during Monday dialysis sessions than on other days of the week. Case patients were nearly twice as likely to have been dialyzed against a 0 or 1.0 mEq/L potassium dialysate on the day of cardiac arrest (17.1 vs. 8.8%). Patients who suffered a cardiac arrest were on average older (66.3 +/- 12.9 vs. 60.2 +/- 15.4 years), more likely to have diabetes (61.8 vs. 46.8%), and more likely to use a catheter for vascular access (34.1 vs. 27.8%) than the general hemodialysis population. Sixteen percent of patients experienced a drop in systolic pressure of 30 mm Hg or more prior to the arrest. Thirty-seven percent of patients who suffered cardiac arrest had been hospitalized within the past 30 days. Sixty percent of patients died within 48 hours of the arrest, including 13% while in the dialysis unit. CONCLUSIONS: Cardiac arrest is a relatively infrequent but devastating complication of hemodialysis. To reduce the risk of adverse cardiac events on hemodialysis, the dialysate prescription should be evaluated and modified on an ongoing basis, especially following hospitalization in high-risk patients. PMID- 11422772 TI - Albumin is the major plasma protein target of oxidant stress in uremia. AB - BACKGROUND: Patients with uremia are exposed to increased oxidative stress. Examination of the oxidation of individual plasma proteins may be useful in establishing specific pathways of oxidative stress in vivo and in determining functional consequences of oxidant stress exposure. We therefore examined oxidative modification of plasma proteins by carbonyl formation using Western blot immunoassay and enzyme-linked immunosorbent assay (ELISA) techniques in patients with chronic renal failure (CRF) and on chronic hemodialysis therapy (HD). METHODS: Plasma was obtained from 25 HD, 20 CRF, and 20 healthy volunteers, derivatized with 2,4 dinitrophenylhydrazine (DNP) and electrophoresed on duplicate 4 to 12% gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels, transferred to nitrocellulose, and stained for DNP for carbonyls and amido black for protein content. Data are recorded as DNP area/protein area and are reported in densitometry units. Total plasma carbonyls were determined by ELISA. RESULTS: Plasma albumin is substantially more oxidized in HD than in healthy volunteers (1.22 +/- 0.14 densitometry units vs. 0.60 +/- 0.08, P = 0.002). There were no significant differences in oxidation of plasma transferrin, immunoglobulin, and fibrinogen in HD versus healthy volunteers. In CRF patients, plasma albumin is more oxidized compared with normal volunteers (1.36 +/- 0.20 densitometry units vs. 0.94 + 0.08, P = 0.09). There were no differences in oxidation of plasma transferrin, fibrinogen, and immunoglobulin in CRF patients versus healthy volunteers. An increased plasma protein carbonyl concentration in CRF patients compared with healthy volunteers was confirmed by ELISA (0.31 +/- 0.07 vs. 0.04 +/- 0.01 nmol/mg protein (P = 0.001). CONCLUSION: Albumin is the major plasma protein target of oxidant stress in CRF and HD patients. PMID- 11422773 TI - In vivo visualization of angiotensin II- and tubuloglomerular feedback-mediated renal vasoconstriction. AB - BACKGROUND: A noninvasive technique to monitor renal microcirculation would be a useful tool for investigation of renal disease and the effects of drugs on the renal system. We have developed a novel, less invasive technique to visualize renal microcirculation in vivo using an intravital tapered-tip (1 mm phi) lens probe (pencil lens-probe) videomicroscopy, which only requires insertion of the probe into superficial renal cortex in situ. METHODS: To assess validity of this technique, the effects of angiotensin II (Ang II) and intrarenal sodium chloride loading (activator of tubuloglomerular mechanism) were examined. The renal microvasculature was successfully visualized and monitored. RESULTS: Administration of Ang II (1, 3, 10 and 30 ng/kg/min) produced a dose-dependent constriction of afferent and efferent arterioles in similar degrees; at 30 ng/kg/min, Ang II elicited 52 +/- 3 (N = 9) and 53 +/- 3% decreases in diameter (N = 9), respectively. The Ang II-induced arteriolar constriction was completely prevented by losartan, an Ang II type 1 (AT1) antagonist. The intrarenal hypertonic saline administration elicited transient increments (from 98 +/- 8 to 122 +/- 7 mL/min, N = 6, P < 0.05), followed by a marked reduction in renal blood flow (RBF; 78 +/- 7 mL/min, P < 0.05). This response was accompanied by prominent constriction of afferent (from 15.0 +/- 1.1 to 8.5 +/- 1.1 microm, N = 6, P < 0.05), but not efferent (from 14.3 +/- 1.2 to 13.8 +/- 1.0 microm, N = 3) arterioles. Furthermore, this response was completely inhibited by furosemide, a tubuloglomerular feedback inhibitor. CONCLUSION: : The intravital pencil lens probe videomicroscopy can be a powerful tool for in vivo observation of renal microcirculation, with intact renal microvascular responses to two important renal homeostatic mechanisms, angiotensin II and tubuloglomerular feedback. PMID- 11422774 TI - Simplified citrate anticoagulation for continuous renal replacement therapy. AB - BACKGROUND: Regional anticoagulation with trisodium citrate is an effective form of anticoagulation for continuous renal replacement therapy (CRRT) for patients with contraindications to heparin. However, because of the metabolic complications of trisodium citrate, it is a complicated technique requiring specialized dialysis solutions. We have designed a simplified protocol for citrate regional anticoagulation for CRRT. METHODS: Two percent trisodium citrate was delivered at 250 mL/h via the prefilter port of a COBE PRISMA device, with the rate adjusted to maintain a postfilter ionized calcium (iCa++) <0.5 mmol/L. A central calcium gluconate infusion was used to maintain a systemic iCa++ at 1.1 mmol/L. A standard dialysate solution consisting of 0.9% saline, KCl 3 mmol/L, and MgSO4 1 mmol/L was delivered at 1000 mL/h. We retrospectively reviewed the outcomes and complications associated with this protoco1 in 29 patients treated from July 1999 to October 1999, evaluating the frequency of clotting of the dialyzer, bleeding complications, citrate toxicity, and patient mortality. RESULTS: The Kaplan-Meier curve for dialyzer survival demonstrated a 61% survival rate at 48 hours. There were no episodes of significant bleeding or citrate toxicity. Seventy-two percent of patients died for reasons unrelated to CRRT. CONCLUSIONS: A CRRT protocol using regional 2% trisodium citrate anticoagulation is not associated with significant bleeding complications or citrate toxicity, and represents a simplified approach compared with previous applications using 4% trisodium citrate. PMID- 11422775 TI - Mast cells find their way to the kidney. PMID- 11422776 TI - Affecting the decline of renal function in diabetes mellitus. PMID- 11422777 TI - Bone remodeling effect of cyclosporine protects against steroid-induced osteopenia. PMID- 11422778 TI - Monogenic hypertension: lessons from the genome. PMID- 11422779 TI - Nitric oxide and the liver. AB - In recent years, the role of nitric oxide (NO) in the pathogenesis of liver disease and its complications has been extensively studied. There remain, however, many areas of controversy. In particular, the effect of NO on vascular function in the systemic circulation and the hepatic microcirculation has received the greatest attention. It has been proposed on the one hand that increased NO synthesis is responsible for the development of the hyperdynamic circulation in cirrhosis, while decreased production of NO within the hepatic microcirculation may be important in the development of parenchymal tissue damage and the onset of portal hypertension. The purpose of this review is to examine the available data concerning the role of NO in liver disease and to discuss some of the controversies and contradictions that surround it. PMID- 11422780 TI - Surgery for biliary atresia. AB - Although the prognosis of biliary atresia has been improved in recent years, particularly in the era of liver transplantation, hepatic portoenterostomy, e.g., the Kasai operation, is still the first line of surgical treatment. Successful hepatic portoenterostomy depends on early diagnosis and operation, adequate operative technique, prevention of postoperative cholangitis, and precise postoperative management. The pathophysiology of the liver and of the intrahepatic bile ducts in this disease is still controversial. PMID- 11422781 TI - Effects of idoxifene and estradiol on NF-kappaB activation in cultured rat hepatocytes undergoing oxidative stress. AB - BACKGROUND/AIMS: Idoxifene is a tissue-specific selective estrogen receptor modulator. Estradiol is a potent endogenous antioxidant, and nuclear factor kappaB (NF-kappaB) is a key transcription factor that induces multiple genes in response to inflammation or oxidative stress. The aim of this study was to explore the inhibitory effects of idoxifene and estradiol on NF-kappaB activation in hepatocytes in a state of oxidative stress. METHODS: Lipid peroxidation was induced in cultured rat hepatocytes by incubation with ferric nitrilotriacetate solution. NF-kappaB activity was evaluated by electrophoretic mobility shift assay. RESULTS: The oxidative stress-induced activation of NF-kappaB and degradation of IkappaB-alpha were maximal at 3-5 h, with an increase in lactate dehydrogenase (LDH) and malondialdehyde (MDA) secretion into the culture medium. Treatment with idoxifene and estradiol inhibited IkappaB-alpha degradation and NF kappaB activation through the attenuation of hepatocyte oxidative bursts and decreased extracellular levels of LDH and MDA. In addition, idoxifene and estradiol inhibited lipid peroxidation in rat liver mitochondria. A potent NF kappaB inhibitor, pyrrolidine dithiocarbamate, prevented NF-kappaB activation by inhibition of IkappaB-alpha degradation and decreased LDH and MDA levels, suggesting that NF-kappaB might be a regulator in a genetic response to increase oxidative stress-induced hepatic injury. CONCLUSIONS: These findings suggest that idoxifene and estradiol function as antioxidants and protect hepatocytes from inflammatory cell injury. PMID- 11422782 TI - Interferon-associated retinopathy in a uniform regimen of natural interferon alpha therapy for chronic hepatitis C. AB - BACKGROUND/AIMS: The development of retinal lesions induced by a uniform regimen of interferon-alpha therapy for chronic hepatitis C was prospectively investigated. METHODS: Eighty-one patients received 6 mega units of natural interferon-alpha intramuscularly daily for 2 weeks and then 3 times a week for 22 weeks. The total dose of interferon-alpha administered was uniformly 478 mega units per patient. Two expert ophthalmologists prospectively examined the patients for retinal lesions before, during and after the therapy. RESULTS: Retinopathy was not found in comparison groups or any of the patients before treatment. In total 34.6% (28/81) of the patients showed cotton-wool spots or minor retinal hemorrhage, or both lesions, during therapy, but these lesions were reversed during or after interferon therapy. The occurrence rates of cotton-wool spots alone, retinal hemorrhage alone and both lesions were 13.6% (11/81), 6.2% (5/81), and 14.8% (12/81), respectively. The appearance of retinopathy did not correlate with patients' background including viral loads and response to the therapy, but was more frequently found in older patients and patients with hypertension and/or diabetes mellitus; disappearance of retinopathy was more prolonged than in patients without these complications. Almost all the lesions appeared 2-4 months after the start of the therapy, and the severity of the lesions did not differ between patients with and without hypertension and/or diabetes mellitus. CONCLUSION: Although it is not clear if interferon-associated retinopathy occurs in a dose-dependent manner, the present study shows a standard pattern of the occurrence of retinopathy in patients with chronic hepatitis C receiving a uniform dosage of natural interferon-alpha. PMID- 11422783 TI - Endothelial nitric oxide synthase and caveolin-1 are co-localized in sinusoidal endothelial fenestrae. AB - BACKGROUND/AIMS: Nitric oxide is synthesized in diverse mammalian tissues by a family of calmodulin-dependent nitric oxide synthases (NOS). Caveolin, the principal structural protein in caveolae, interacts with endothelial NOS leading to enzyme inhibition in a reversible process modulated by Ca++-calmodulin. The aim of the present study was to clarify the ultrastructural localization of eNOS and caveolin-1 in hepatic sinusoidal endothelium by an electron immunogold method. METHODS: Male Wistar rats were used. Liver tissues and hepatic sinusoidal endothelial cells isolated from rat livers by collagenase infusion were studied. For immunohistochemistry, liver specimens were reacted with anti-eNOS or anti caveolin-1 antibody. The ultrastructural localization of eNOS or caveolin-1 was identified by electron microscopy using an immunogold post-embedding method. RESULTS: Immunohistochemical studies using liver tissues localized endothelial NOS in hepatic sinusoidal lining cells, portal veins and hepatic arteries; and caveolin-1 in sinusoidal lining cells, bile canaliculi, portal vein and hepatic arteries. Immunogold particles indicating the presence of eNOS and caveolin-1 were demonstrated on the plasma membrane of sinusoidal endothelial fenestrae in liver tissue and also in isolated sinusoidal endothelial cells. CONCLUSION: Endothelial NOS and caveolin are co-localized on sinusoidal endothelial fenestrae, suggesting that interaction of the two may modulate cellular regulation of NO synthesis. PMID- 11422784 TI - Efficacy and safety of an intravenous monoclonal anti-HBs in chronic hepatitis B patients. AB - BACKGROUND/AIMS: In this study the safety and efficacy of a monoclonal anti-HBs, Tuvirumab (Mab), were investigated. Tuvirumab is a human monoclonal antibody recognizing the stable 'a'-determinant of the HBsAg. METHODS: We included ten chronic hepatitis B patients: four received monotherapy, and six combination therapy with interferon alpha 2b. RESULTS: Because the development of insoluble [HBsAg-HBsAb] complexes led to adverse events, the Mab dose had to be reduced in seven patients. In nine patients treatment was stopped prematurely because of lack of efficacy, i.e. neutralization of HBsAg in serum. However, temporary HBsAg levels were reduced by at least 50% in all patients; in three patients receiving combination therapy, background levels of HBsAg in serum were reached. A loss of serum HBV-DNA was seen in three patients in the combination group, followed by HBeAg seroconversion in two patients. CONCLUSIONS: We conclude that Mab was not effective in achieving primary efficacy as assessed by neutralization of circulating HBsAg. Whether a combination of Mab with an antiviral agent that reduces the HBsAg load--and therefore minimizes the risk of adverse events--may result in clinical efficacy should be investigated. PMID- 11422785 TI - Cholangiopathy after short-term administration of piperacillin and imipenem/cilastatin. AB - We describe a patient who suffered from intestinal perforation after abdominal trauma. Perioperatively, he was treated with a single dose of piperacillin and 9 doses of imipenem/cilastatin over 3 days. The patient was discharged 5 days after surgery in good clinical condition and with normal liver values except for a marginal elevation of alanine aminotransferase. Two weeks after discharge, he developed fatigue, fever and pruritus, necessitating rehospitalization. He was jaundiced and had elevated alkaline phosphatase and transaminases. After exclusion of an intra-abdominal fluid collection, a vascular problem, and infectious or autoimmune liver disease, a liver biopsy was performed. The biopsy revealed centrizonal bilirubinostasis, a portal infiltrate rich in eosinophils and cholangitis. Lymphocyte transformation tests for piperacillin and imipenem/cilastatin were positive, suggesting an immunological mechanism for the observed hepatopathy. Cholestasis gradually decreased but was detectable for several weeks. The patient had a full clinical and biochemical recovery after 3 months. We conclude that short-term therapy with piperacillin, imipenem/cilastatin or the combination of these drugs can lead to the same type of hepatopathy as described for amoxycillin/clavulanic acid or antistaphylococcal penicillins. Liver biopsy and positive lymphocyte transformation are compatible with an immunological mechanism. PMID- 11422786 TI - Concomitant inflammatory pseudotumor of the liver and spleen. AB - We report the case of a 53-year-old man with inflammatory pseudotumor (IPT) of the liver and spleen. This concomitant association has rarely been reported. The patient presented with a hypoechoic mass in the liver and a clinical picture of recurrent sepsis; hematochemical exams and imaging data were nonspecific. Antibiotic therapy improved the clinical course, but did not resolve it definitively. After 50 days of therapy, as the hepatic mass decreased a similar lesion appeared in the spleen. The final diagnosis was made on splenectomy and an intra-operative biopsy of the residual liver lesion. The diagnostic problems encountered in this very rare association of IPT of the liver and spleen were similar to those for isolated IPT in the respective single organ sites. After 15 months of follow-up, the patient is in good health and no recurrence of symptoms or masses has been observed. PMID- 11422787 TI - Effect of bezafibrate in primary biliary cirrhosis: a pilot study. PMID- 11422788 TI - Cell differentiation and apoptosis of monocytic and promyelocytic leukemia cells (U-937 and HL-60) by tryptanthrin, an active ingredient of Polygonum tinctorium Lour. AB - Tryptanthrin, a bioactive ingredient of Polygonum tinctorium Lour., is a member of the Indigo plant family and has potent cytocidal effects on various human leukemia cells in vitro. At low concentrations, tryptanthrin enhanced the expression of cell differentiation (CD) markers in human monocytic (U-937) and promyelocytic (HL-60) leukemia cells indicative of differentiation to monocytes/macrophages. Furthermore, nitroblue tetrazolium (NBT) reductive and alpha-naphthyl butyrate esterase (NBE) activities were markedly increased after treatment. Tryptanthrin was more potent than dimethyl sulfoxide (DMSO) at inducing U-937 cell differentiation into monocytes/macrophages. After treatment with higher concentrations of tryptanthrin for 24 h, cytoplasmic vacuolation and destruction of mitochondria were observed. The leukemia cells died via apoptosis 48 h after treatment. Cytoplasmic vacuolation and apoptotic changes correlated with the dysfunction of mitochondria. Electron microscopic observations revealed marked swelling and destruction of mitochondria after exposure of the leukemia cells to tryptanthrin. Exposure to tryptanthrin enhanced Fas-induced apoptosis and increased caspase-3 activity before induction of apoptosis. These results show that low concentrations of tryptanthrin can induce differentiation of leukemia cells but higher concentrations will kill leukemia cells through apoptosis, possibly through a caspase-3/Fas antigen pathway. PMID- 11422789 TI - Vascular amyloid of unknown origin and senile transthyretin amyloid in the lung and gastrointestinal tract of old age: histological and immunohistochemical studies. AB - The histological and immunohistochemical characteristics and the incidence of amyloid deposits in the tissues of the lung and gastrointestinal tract were investigated in 64 autopsied individuals who were 80 years and older (age range: 80-92 years; mean: 83.3 years). Immunohistochemical examination was performed with antibodies against amyloid A, transthyretin, immunoglobulin lambda and kappa light chain amyloid fibril proteins, beta2-microglobulin, beta protein, apolipoprotein AI, apolipoprotein AII, atrial natriuretic peptide, apolipoprotein E, and amyloid P component. Transthyretin amyloid fibril protein (ATTR) deposits were observed in five cases (7.8%). Gastrointestinal amyloid deposits of unknown origin were observed in the veins of the gastrointestinal tract in 26 cases (40.6%). This amyloid was regarded as portal amyloid with respect to distribution pattern. Pulmonary vascular amyloid deposits of unknown origin were observed in 12 cases (18.8%). These amyloid deposits were found mainly in medium-sized veins in the lungs and did not react with any antibodies against amyloid fibril proteins except apolipoprotein E and amyloid P component. Eleven of the 26 cases (42.3%) showing portal amyloid also showed pulmonary vascular amyloid of unknown origin. The pulmonary vascular amyloid deposits were similar to the portal amyloid deposits with respect to their morphological features and their relation to elastic fibers in the vessels. Further morphological investigation and biochemical analysis of the pulmonary vascular amyloid and portal amyloid will resolve questions of their origins and relation. PMID- 11422790 TI - Immunohistochemical expression of gonadotropin releasing hormone receptor in human breast carcinoma. AB - Gonadotropin releasing hormone (GnRH) analogs can cause regression of hormone dependent breast carcinomas via the specific GnRH receptor (GnRH-R). In an attempt to obtain a better understanding of GnRH actions in human breast carcinoma, the expression of GnRH-R was examined immunohistochemically in 58 invasive ductal carcinomas and correlated with various clinicopathological parameters. GnRH-R was immunolocalized in the cytoplasm of carcinoma cells in 37 of 58 invasive ductal carcinoma cases (64%). Immunoreactivity for GnRH-R was also detected focally in the cytoplasm of morphologically normal glandular epithelia adjacent to the carcinoma. A significant correlation was observed between the immunohistochemical expression of GnRH-R and estrogen receptor labeling index (LI; P = 0.030) or progesterone receptor LI (P = 0.0074). There was a significant inverse correlation between GnRH-R immunoreactivity and Ki-67 LI (P = 0.012). No significant correlations were detected between GnRH-R and other clinicopathological parameters, including patient age, menopausal status, stage, tumor size, lymph node status, histological grade and prognosis. This study indicates that GnRH-R is widely distributed in human breast carcinoma cells and regulates GnRH actions locally. Breast carcinomas positive for GnRH-R maintain some hormonal regulatory mechanisms, and GnRH actions may lead to a low proliferative rate in human breast carcinoma. PMID- 11422791 TI - Relationship between skeinoid fibers and stromal matrix in gastrointestinal stromal tumors: morphometric analysis with quick-freezing and deep-etching method. AB - Our previous study of a gastrointestinal autonomic nerve tumor with skeinoid fibers (SF) using the quick-freezing and deep-etching method, suggested that the distance between one radix and a neighboring radix (DRNR) in pre-existing meshwork structures around the tumor cells is consistent with the periodicity of the SF. Therefore, measurement of the DRNR in the meshwork could clarify the significance of the pericellular matrix for SF development. In the present study, we analyzed the meshwork in three cases of gastrointestinal stromal tumor (GIST), which showed different immunohistochemical stainings, but confirmed to have smooth muscle differentiation (SMD) by immunohistochemistry and/or electron microscopy. The DRNR from the three cases of GIST showed similar histogram patterns (a peak of 20-30 nm, mean values of 28.02, 25.74 and 26.45 nm), which were significantly shorter than the periodicity of SF (a peak of 40-45 nm, mean value of 42.14). Although we need further studies with additional GIST cases, we speculate that the pericellular matrix of GIST with SMD is not suitable for SF development. PMID- 11422792 TI - Histopathological study on mechanism and background of tumor-forming pancreatitis. AB - Fifteen cases of tumor-forming pancreatitis, detected as tumors by diagnostic imaging or by physical examination were histologically examined. Eleven of the 15 patients were heavy drinkers. Tumorous lesions were located in the head of the pancreas in 11 cases and in the body or tail of the pancreas in four cases. Macroscopic examination revealed tumorous swelling or sclerotic appearance in the pancreatic tissue. Histologically, these lesions showed tumorous swelling with (n = 12) or without (n = 3) a background of chronic pancreatitis. In the former, the tumorous lesions consisted of extensive fibrosis, including necrosis or abscesses, stones and reparative granulation tissue, and there was a successive transition to the surrounding chronic pancreatitis pattern. The latter three tumorous lesions presented with inter- and intralobular fibrosis with lymphoid hyperplasia or lymphoplasmacytic infiltration and were adjacent to normal pancreatic tissue. Therefore, tumor-forming pancreatitis shows at least two distinct types: a reparative tumorous swelling with a background of chronic pancreatitis, which is considered to have given rise to the tumor at some stage; and a lymphoid and fibrous proliferation in normal pancreatic tissue, which is considered to represent an autoimmune-related disease process. PMID- 11422793 TI - Epstein-Barr virus infection, drug resistance and prognosis in Korean T- and NK cell lymphomas. AB - T-cell lymphomas are a biologically heterogeneous group of diseases with varying clinical presentations and outcomes. We tried to understand the effect of Epstein Barr virus (EBV) on lymphogenesis, prognostic factors and drug resistance of T cell lymphomas, and to establish their relationship with international prognostic factors. Formalin-fixed paraffin-embedded tissue sections from 35 patients (12 women and 23 men) with T-cell lymphomas were examined to detect the presence of EBV using RNA in situ hybridization for EBV-encoded small nuclear RNA (EBER) 1/2 and immunohistochemical stain for latent membrane protein (LMP)-1. We also tried to establish the expression of p53 and P-glycoprotein (P-gp) using immunohistochemistry. The distribution according to the subgroup was: two T lymphoblastic lymphomas, 13 NK/T-cell lymphomas, one angioimmunoblastic T-cell lymphoma, 17 peripheral T-cell lymphomas, unspecified, and two anaplastic large cell lymphomas. The EBER was detected in 15 of 35 T-cell lymphomas (42.9%) and among these it was detected in five of 17 nodal lymphomas (29.4%) and 10 of 18 extranodal lymphomas (55.6%). There was close correlation between EBER positivity and NK/T-cell lymphoma (P = 0.032). Expression of LMP was found in a proportion of tumor cells in seven of the 15 EBER-positive cases (46.7%). There was no correlation between EBER expression and complete response (CR rate), but coexpression of EBER and p53 was associated with treatment failure (P = 0.047). The 18 patients (51.4%) with p53 expression had significantly poorer outcomes compared with the 17 patients without p53 expression (CR rate, P < 0.0005; overall survival, P = 0.0102). Twenty of 35 patients (57.1%) were positive for P gp expression. P-gp expression was significantly associated with treatment failure (P = 0.001) and overall survival (P = 0.0089). Seventeen of 35 patients (48.6%) treated with systemic chemotherapy or radiation therapy achieved a CR after initial treatment. When the prognostic factors were grouped using the international prognostic index, the CR rate was 58.8% for the low risk group, 50.0% for the low-intermediate risk group, 14.3% for the high-intermediate risk group, and 0% for the high risk group. In conclusion, high incidence of EBV was detected among Korean patients with T-cell lymphomas. Our study supports the prediction that patients who express p53 and P-gp have a poorer prognosis than those who do not and this should be considered when treatment strategies for individual patients are selected. PMID- 11422794 TI - Discrepancies between Gleason scores of needle biopsy and radical prostatectomy specimens. AB - The purpose of this study was to determine the accuracy of Gleason scores in prostate needle biopsy diagnosis and to investigate factors affecting the accuracy of the tumor grade. A single pathologist reviewed 116 sets of prostate cancer biopsies and radical prostatectomy specimens. The following factors were examined to determine their effect on the accuracy of the biopsy Gleason scores: (i) relative tumor differentiation; (ii) pathological stage; (iii) amount of tissue in the biopsy specimen; (iv) amount of cancer tissue in the biopsy specimen; (v) tumor heterogeneity; (vi) clinical findings (prostate specific antigen value and digital rectal examination); and (vii) interobserver variability. In 53 cases the Gleason score of biopsy specimens was identical to the score of prostatectomy specimens (45.7%). Fifty-four cases (46.6%) of biopsy specimens were undergraded. The most common discrepancy was diagnosis of well differentiated carcinoma in the biopsy but diagnosis of moderately differentiated tumor in the corresponding prostatectomy specimen. This discrepancy occurred when the amount of tumor in the biopsy was 3 mm or less. Biopsy and prostatectomy results showed less agreement when the original biopsy tumor grade rendered by nine different pathologists was used, suggesting that interobserver variability can adversely affect the accuracy of tumor grade. Clarifying the histologic criteria for distinguishing each grade, especially between Gleason grades 2 and 3, is important for accurate grading. PMID- 11422795 TI - Imported malaria in a Japanese male: an autopsy report. AB - Fatal cases of malaria are rare in Japan. We report a case of a 47-year-old Japanese man with Plasmodium falciparum malaria. The patient was examined because of fever and headache after a trip to Africa. He was diagnosed with malaria. Chemotherapy begun on day three decreased the percentage of infected red blood cells (RBC) from 25% to 2%, but the patient fell into coma on the same day. The patient was considered brain dead for 3 days before he died, and he was autopsied on day nine. Brain hemispheres were preserved and swollen with meningeal congestion. The ventral area of the pons and medulla oblongata were softened, and the tonsils of the cerebellum were softened and herniated. The spleen was blackish, enlarged and showed a small infarction. The liver was yellowish and enlarged. Many infected RBC were seen in the capillaries of the brain and malaria pigments were seen in the spleen and liver. DNA of P. falciparum was detected by polymerase chain reaction from paraffin-embedded brain materials, however, the DNA could not be detected in other organs. Besides malaria, the patient had latent primary thyroid cancer, which was a small and invasive papillary carcinoma. PMID- 11422796 TI - An autopsy case of pancreatic and ectopic nesidioblastosis. AB - We report an autopsy case of pancreatic and ectopic nesidioblastosis. A five month-old Japanese girl was born at 35 weeks gestation, and showed clinical symptoms of hyper-insulinemic hypoglycemia before death. At autopsy a tumorous nodule was observed at the portion of the jejunum, 90 cm from Treitz's ligament. The nodule measured 30 x 20 x 20 mm. The ectopic pancreas, also revealed nesidioblastosis histologically. Immunohistologically, both nesidioblastoses were stained positive for chromogranin A, insulin, glucagon and somatostatin. The proliferating cell nuclear antigen (PCNA) and Ki-67 indices were less than 4% in the nesidioblastosis. To our knowledge, this is the first reported case of nesidioblastosis demonstrating proliferating activity with PCNA and Ki-67, and is the third reported case of nesidioblastosis arising in the pancreas and ectopic pancreas. PMID- 11422797 TI - Ovarian carcinoma recurring as carcinosarcoma. AB - Malignant mixed mesodermal tumor is a rare tumor of the ovary and its histogenesis is controversial. We report the case of an ovarian tumor that seemed to be a pure carcinoma and recurred as a carcinosarcoma, and suggest a possible histogenesis for this kind of tumor. The patient was a 62-year-old Japanese woman. The primary tumor was confined to the right ovary and was a histologically poorly differentiated endometrioid adenocarcinoma with focal squamous differentiation. The tumor recurred as peritoneal dissemination 9 months later showing a histological appearance of carcinosarcoma of heterologous type. The recurrent tumor also contained intermingled foci of similar histology as the primary tumor. The carcinomatous component of the recurrent tumor showed more obvious differentiation to adenocarcinoma with increased expression of epithelial markers compared to the primary tumor. Epithelial membrane antigen was positive also in a few cells of the sarcomatous component, which implies that this tumor had features of metaplastic carcinoma. The DNA ploidy pattern of the primary ovarian tumor was diploid, while an additional aneuploid subpopulation appeared in the recurrent tumor. These findings suggest the possible histogenesis of carcinosarcoma of the ovary as progression and clonal evolution of endometrioid adenocarcinoma. PMID- 11422798 TI - Primary synovial sarcoma of the kidney: Report of a case confirmed by molecular detection of the SYT-SSX2 fusion transcripts. AB - We describe an unusual case of primary synovial sarcoma of the kidney. A 47-year old woman had a tumor massively replacing the right kidney. There were no primary extrarenal neoplastic lesions. Microscopically, the tumor was composed of a cellular proliferation of relatively uniform spindle-shaped cells having atypical spindle or oval nuclei arranged in fascicles with tumor necrosis, without epithelial areas. Immunohistochemically, a small number of the tumor cells were positive for epithelial markers such as cytokeratin and epithelial membrane antigen. The SYT-SSX2 fusion transcripts were detected by a reverse transcription polymerase chain reaction (RT-PCR) using RNA extracted from formalin-fixed, paraffin-embedded tissue. ETV6-NTRK3 fusion gene transcripts that result from t(12; 15)(p13;q25), which is characteristic of cellular congenital mesoblastic nephroma, were not demonstrated. To our knowledge, this is the ninth case of primary renal synovial sarcoma. This case report indicates that synovial sarcoma should be taken into account for the differential diagnosis of renal spindle cell tumors and the molecular assay detecting the SYT-SSX fusion transcripts is useful for the final diagnosis of synovial sarcoma arising in an unusual location. PMID- 11422799 TI - Histologically low-grade dedifferentiated liposarcoma of the retroperitoneum. AB - A low-grade dedifferentiated liposarcoma in the retroperitoneum of a 52-year-old woman is described. The excised specimens contained six nodules of lipoma-like well-differentiated liposarcoma and a nodule of dedifferentiated liposarcoma. The latter was composed predominantly of loosely arranged, benign-appearing spindle cells and fat cells. A small number of cells with irregularly shaped nuclei were scattered. There were no mitotic figures. The fat cells showed slight variation of size and shape, and a few multivacuolated lipoblasts were found. The spindle cell areas occupied approximately 60% of the tumor. The stroma was somewhat fibrous and myxoid and no dense collagenous matrix was found. The stroma vascularity was not prominent. Immunohistochemically, the spindle cells were positive for vimentin but negative for S-100 protein, desmin, muscle actin, and alpha-smooth muscle actin. Follow up for 5 months showed no evidence of recurrence or metastasis. The tumor, in which the benign-appearing spindle cell component was predominant, was considered to be a low-grade dedifferentiated liposarcoma. Close and long-term follow up is required. In retroperitoneal lipoma like well-differentiated liposarcomas, spindle cell components like the present tumor, which represent dedifferentiation, should not be overlooked. PMID- 11422800 TI - Intravenous leiomyomatosis of the uterus with multiple pulmonary metastases associated with large bullae-like cyst formation. AB - We present a case of uterine intravenous leiomyomatosis associated with multiple pulmonary metastases with bullae-like cystic change. A 53-year-old woman who had undergone hysterectomy 5 years previously underwent an operation for multiple pulmonary nodules with bullae formation. After resection of several large bullae, a subsequent extirpation of the pulmonary nodules was performed, and a pathological examination showed multiple leiomyomatous nodules with occasional cystic change. A review of the previous slides of the uterus and immunohistochemical analysis of the proliferating ability using anti-Ki-67 and anti-proliferating cell nuclear antigen (PCNA) antibodies were performed. Proliferating cells of the uterus had very few mitotic figures for their high cellularity, and the labeling indices of Ki-67 and PCNA indicated very low levels in both uterine neoplasm and pulmonary nodules. From these findings, an intravenous leiomyomatosis associated with multiple pulmonary metastases was diagnosed. PMID- 11422801 TI - Thyrocyte integration, and thyroid folliculogenesis and tissue regeneration: perspective for thyroid tissue engineering. AB - The thyroid gland is composed of many ball-like structures called thyroid follicles, which are supported by the interfollicular extracellular matrix (ECM) and a capillary network. The component thyrocytes are highly integrated in their specific structural and functional polarization. In conventional monolayer and floating culture systems, thyrocytes cannot organize themselves into follicles with normal polarity. In contrast, in 3-D collagen gel culture, thyrocytes easily form stable follicles with physiological polarity. Integration of thyrocyte growth and differentiation results ultimately in thyroid folliculogenesis. This culture method and subacute thyroiditis are two promising models for addressing mechanisms of folliculogenesis, because thyroid-follicle formation actively occurs both in the culture system and at the regenerative phase of the disorder. The understanding of the mechanistic basis of folliculogenesis is prerequisite for generation of artificial thyroid tissue, which would enable a more physiological strategy to the treatment of hypothyroidism caused by various diseases and surgical processes than conventional hormone replacement therapy. We review here thyrocyte integration, and thyroid folliculogenesis and tissue regeneration. We also briefly discuss a perspective for thyroid tissue regeneration and engineering. PMID- 11422802 TI - Spontaneous occurrence of chronic non-suppurative destructive cholangitis and antimitochondrial autoantibodies in MRL/lpr mice: possible animal model for primary biliary cirrhosis. AB - MRL/MP mice bearing the lymphoproliferative gene lpr (known as MRL/MP-lpr/lpr or MRL/Ipr mice) are known to spontaneously develop severe autoimmune diseases such as glomerulonephritis, arteritis and arthritis at an early stage of their life. They have also been reported to develop severe sialadenitis, suggesting that this mouse could be a model for Sjogren's syndrome. Primary biliary cirrhosis, an autoimmune disease characterized by chronic non-suppurative destructive cholangitis and the occurrence of antimitochondrial antibodies, is frequently associated with Sjogren's syndrome. In this study, we examined whether cholangitis and/or antimitochondrial antibodies occur in this mouse model, using more than 100 young and old MRL/Ipr mice. We frequently found portal inflammation associated with cholangitis of small intrahepatic bile ducts, especially in older mice. There was also infiltration of inflammatory cells (monocytes) as well as CD4-positive T cells. Epithelioid granuloma and bile-duct loss were also occasionally found. These histological features resemble primary biliary cirrhosis. In addition, antimitochondrial antibodies were shown by immunocytochemistry to be present in the sera of MRL/Ipr mice. There is currently no established animal model for primary biliary cirrhosis. Therefore, further studies on MRL/Ipr mice, with respect to pathogenesis of primary biliary cirrhosis, are warranted. PMID- 11422803 TI - 3-D analysis of vascular and capsular invasion in thyroid follicular carcinoma. AB - To clarify the mechanism of vascular invasion (VI) and capsular invasion (CI) in thyroid follicular carcinoma (FC), 3-D reconstruction of tumor tissues, vessels and capsules was performed from serial sections in five FC and three follicular adenomas (FA). Outflow veins in the lesions were well developed under the capsule, narrowed just beneath the capsule, and dilated within the capsule. Both VI and CI were observed at the poststenotic intracapsular dilated part of outflow veins. The tumor tissue extended along the intracapsular horizontal veins which flowed in parallel with the capsule in VI, or along the vertical veins which directly flowed outside the capsule in CI. There was tumor tissue in the subendothelial layer of the outflow veins within the capsule. Intravascular tumor nests and nodular lesions within the capsule continued from the main tumors. It can be concluded from these results that most VI and CI may not be true invasions but pre-invasive lesions. PMID- 11422804 TI - Comparative immunochemical study of lectin-binding sites and cytoskeletal filaments in static and reactive mesothelium and adenocarcinoma. AB - In cytological preparations, reactive mesothelial cells (RMC) in serous effusions are sometimes difficult to distinguish from adenocarcinoma cells (AC). RMC and AC can be distinguished by lectin-binding patterns, but the pattern of binding of lectins to normal mesothelium is not well defined. We investigated the expression of cytoskeletal filaments, cytokeratin (CK) and vimentin (VM), and the cell surface binding pattern of 10 lectins (HPA, SBA, ABA, DSA, PNA, RCA-I, UEA-I, LTA, WGA and ConA) in the serosa of 48 adenocarcinoma specimens. We also investigated the usefulness of six lectins (HPA, SBA, RCA-I, UEA-I, LTA and WGA) in identification of RMC and AC in 16 serous effusions. DSA reactivity was significantly higher (P < 0.05) in static mesothelial cells (SMC) than in RMC. Reactivity for LTA and ConA was significantly lower (P < 0.05) in SMC than in RMC. Anti-CK and anti-VM immunoreactivity was always positive in RMC and almost negative in SMC. In serous effusions, HPA, SBA and UEA-I binding was evident in 100, 88 and 81% of AC, respectively. Little to no binding of HPA, SBA or UEA-I was detected in RMC. Our results suggest that the morphological differences between SMC and RMC are likely to be due to differences in cytoskeletal composition, with accompanying changes in cell-surface lectin-binding patterns. HPA, SBA and UEA-I are likely to be useful markers for identification of RMC and AC in cytology. PMID- 11422805 TI - Expression of p53 protein as a prognostic indicator of reduced survival time in diffuse-type gastric carcinoma. AB - To determine whether p53 expression is different in intestinal and diffuse types of gastric carcinoma, we investigated p53 immunohistochemical expression in 178 primary gastric carcinomas. Overexpression of p53 was observed in 50 out of 100 intestinal-type tumors (50.0%) and in 27 out of 78 diffuse-type tumors (34.6%). A significant difference was found in the timing of p53 overexpression between the two types of carcinomas. Overexpression of p53 occurred often in the early stage of intestinal-type tumors, and there was no significant difference in expression between early and advanced cancers. In contrast, p53 overexpression did not occur often in the early stage of diffuse-type tumors, but it increased progressively as the tumor advanced. Analysis of patient survival revealed that p53 overexpression correlates significantly with a poor prognosis in diffuse-type gastric carcinoma (P = 0.003) but not in intestinal-type. Multivariate analysis showed that only pathological stage was an independent prognostic indicator. Our results suggest that p53 overexpression plays a different role in tumor carcinogenesis and progression of these two types of gastric cancers. PMID- 11422806 TI - Methylation of the hMLH1 promoter in multiple gastric carcinomas with microsatellite instability. AB - Hypermethylation of the hMLH1 promoter is observed in the majority of sporadic gastric carcinomas with high frequency microsatellite instability (MSI), and it contributes to the genesis of MSI-positive gastric carcinoma. Multiple gastric carcinoma is known to have a higher frequency of MSI positivity than single gastric carcinoma. However, the molecular basis of MSI in these tumors remains obscure. We investigated the role of hMLH1 promoter hypermethylation in the genesis of multiple gastric carcinoma with MSI. We analyzed 33 tumors from 15 patients with multiple gastric carcinoma (12 double tumors and three triple tumors) for MSI, expression of hMLH1 and hMSH2, and hypermethylation of hMLH1 and hMSH2 promoters. High frequency MSI was found in seven out of 33 tumors (21%) in five out of 15 patients (33%). All of the tumors with high frequency MSI had a lack of hMLH1 expression, with the presence of hMSH2 expression, while all the tumors with no MSI or low frequency MSI were positive for both hMLH1 and hMSH2. All of the tumors with no expression of hMLH1 had hMLH1 hypermethylation, whereas hMLH1 hypermethylation was observed in two out of 26 (8%) tumors with no or low frequency MSI. None of the tumors showed hMSH2 hypermethylation. These results suggest that epigenetic changes in the hMLH1 promoter account for the genesis of multiple gastric carcinoma with high frequency MSI. PMID- 11422807 TI - Prognostic significance of neuroendocrine differentiation, proliferation activity and androgen receptor expression in prostate cancer. AB - Androgen, acting via the androgen receptor (AR), is associated with the development and progression of prostate cancer. Anti-androgen therapy is widely used to manage prostate cancer. However, the conversion of the tumor from a hormone-sensitive to a hormone-insensitive status causes such therapy to fail. Several mechanisms have now been put forward for this conversion, including neuroendocrine (NE) differentiation of the tumor cells. In this study, we evaluated the prognostic significance of tumor-cell proliferation activity, NE differentiation and AR expression. Formalin-fixed, paraffin-embedded sections were prepared from 42 patients with adenocarcinoma of the prostate. Using antibodies to AR, the Ki-67 antigen (MIB-1), chromogranin A and synaptophysin, immunohistochemical expression of AR, tumor proliferation activity and NE differentiation were analyzed. Our study revealed that AR expression was significantly lower in adenocarcinoma (52.2 +/- 27.1%) than in non-tumorous prostate tissue (68.3 +/- 18.3%; P < 0.001). NE differentiation was found in 50% of the tumors, which was correlated with the Gleason score (P < 0.05). An univariate analysis revealed a significant correlation between progression-free survival with both AR expression (P < 0.01) and proliferation activity (P < 0.001). NE differentiation was not a prognostic factor in this study. PMID- 11422808 TI - Histological diversity of vasculitic lesions in MPO-ANCA-positive autopsy cases. AB - To investigate the variety of histological features of vasculitic lesions in myeloperoxidase-specific antineutrophil cytoplasmic antibody (pANCA)-related vasculitis, retrospective pathological analysis was done on 13 autopsy cases, collected from 1990 to 1998 at five hospitals. These cases were classified into three groups: (i) pulmonary-renal syndrome characterized by capillaritis of lung and glomeruli with occasional small-vessel arteritis and/or phlebitis; (ii) glomerular capillaritis without pulmonary involvement associated with significant small-vessel arteritis; and (iii) extensive distribution of small-vessel arteritis with no capillary involvement. The results suggest that pANCA-related vasculitis encompasses a wide variety of vasculitic syndromes, including pulmonary-renal syndrome, microscopic polyarteritis nodosa, and classic polyarteritis nodosa. pANCA may contribute to pathogenesis in all of these cases. PMID- 11422809 TI - Three autopsied cases of cystic fibrosis in Japan. AB - The incidence of cystic fibrosis (CF) is very rare in Japanese, while it is frequent in Caucasians. Here we report on three Japanese cases of CF. One of the patients had consanguineous parents. All three patients initially developed meconium ileus, and hepatobiliary and pancreatic changes became more severe as age increased. The DeltaF508 mutation, the most frequent mutation associated with CF in Caucasians, was not found in these patients. To evaluate the relationship between the severity of hepatic lesions and a history of meconium ileus, we examined hepatic lesions in the present three cases, and we reviewed 22 Japanese autopsied cases of CF in the literature. No correlation was found between the incidence of biliary cirrhosis and a history of meconium ileus, because the cases with meconium ileus showed a high mortality during the neonatal period, before biliary cirrhosis developed. The high incidence of meconium ileus in Japanese CF patients may relate to a clinically severe phenotype and reflect a different genetic background between Caucasians and Japanese. PMID- 11422810 TI - Absence of BAT-26 instability in gastric intestinal metaplasia. AB - BAT-26 instability, a sensitive marker for the high-frequency microsatellite instability (MSI-H) phenotype, was analyzed in samples of gastric cancer and in adjacent intestinal metaplastic mucosae. Although all MSI-H gastric cancer samples showed BAT-26 instability, as assessed using 12 dinucleotide microsatellite markers, BAT-26 instability was not found in the adjacent intestinal metaplastic mucosa in any of the samples. PMID- 11422811 TI - Familial amyloidotic polyneuropathy (ATTR Val30Met) with widespread cerebral amyloid angiopathy and lethal cerebral hemorrhage. AB - We report an autopsy case of familial amyloidotic polyneuropathy (FAP) with cerebral hemorrhage. A 38-year-old woman with a typical FAP pedigree started developing severe diarrhea and sensori-motor polyneuropathy at the age of 28 years; autonomic nervous system, heart and renal dysfunction manifested themselves in the following years. Genetic analysis revealed a single amino acid substitution at codon 30 of transthyretin (ATTR Val30Met). Ten years after her initial symptoms, the patient died of a sudden convulsive attack and respiratory failure. Autopsy revealed lethal cerebral hemorrhages and uremic lungs. Histochemical and immunohistochemical analyses revealed TTR-derived amyloid protein in every tissue examined, particularly in glomeruli and peripheral vessels. Severe meningo-cerebrovascular amyloidosis was also detected. Because uremia causes oxidative damage to the vascular system and amyloid formation is closely associated with oxidative stress, it is possible that uremic endothelial damage facilitated an unusual cerebral amyloid deposition. In typical FAP (ATTR Val30Met), cerebral amyloid angiopathy does not usually have clinical manifestations. However, cerebral amyloid angiopathy should be considered to explain FAP symptoms when some risk factors such as uremic vascular damage are accompanying features. PMID- 11422812 TI - Giant intracranial granular-cell tumor arising from the abducens. AB - We present a case of an adolescent with a giant intracranial granular-cell tumor, who presented with diplopia and worsening visual acuity. The tumor measured 8.2 cm at the maximum diameter and occupied the right middle cranial fossa, with intrasellar and suprasellar involvement. It was composed of solid nests of oval or polygonal eosinophilic cells with a focal pseudo-papillary configuration. Pituitary macroadenoma was suspected from examination of the intraoperative frozen section, but immunohistochemical and ultrastructural findings were consistent with granular-cell tumor, and they suggested the origin to be the peripheral nerve sheath. Considering that abducens palsy was the first clinical manifestation, the tumor was thought to arise from abducens. From histological findings (focal cellular spindling, mitotic activity and a relatively high Ki-67 labeling index [10.1%]) and according to Fanburg-Smith's criteria, the tumor was suspected to have low-grade malignant potential. Granular-cell tumor should be included in the differential diagnosis of parasellar tumors, even in young patients or patients with large lesions. Both immunohistochemical and electron microscopic studies are essential for diagnosis. PMID- 11422813 TI - Composite glandular-endocrine cell carcinoma of the common bile duct. AB - A rare case of composite glandular-endocrine cell carcinoma of the common bile duct is presented. Histologically, this tumor consisted of adenocarcinoma and small-cell neuroendocrine carcinoma, with a transition between the two components. The two distinct areas of the tumor were immunohistochemically different, whereas the transitional zone exhibited characteristics of both areas. These features suggest that the tumor arose from a multipotential stem cell. Although it has been reported that the presence of neuroendocrine differentiation in carcinomas indicates a poor prognosis, the patient in the present case was well at the time of writing this report. This may be due to the fact that adenocarcinoma, which characteristically has a low proliferative activity, constituted the majority of the tumor. PMID- 11422814 TI - Pediatric kidney transplants--who shall donate? PMID- 11422815 TI - Adaptation of split liver grafts in pediatric patients. PMID- 11422816 TI - Varicella-zoster infection in pediatric solid-organ transplant recipients: a hospital-based study in the prevaricella vaccine era. AB - We reviewed 58 cases of varicella-zoster infection that occurred between 1988 and 1998 in 47 pediatric solid-organ transplant recipients. The median age of patients at the time of admission with varicella-zoster infection was 8.0 yr (range 1-17 yr). The median interval between transplantation (Tx) and varicella zoster virus (VZV) infection was 1.6 yr (range 0.06-9.3 yr). Varicella infection occurred at a rate of one case for every seven transplant recipients. Among the 58 cases of VZV infection, 53% were varicella while 47% were herpes-zoster. Varicella infection occurred despite treatment with varicella-zoster immune globulin (VZIG) in 17 of 31 cases of varicella infection. However, the disease was generally mild with severe disease occurring in only two patients. One patient (1.7%) died as a result of bacterial sepsis. There was no significant relationship between VZV infection and specific immune suppressants. Episodes of rejection were more likely to be temporally associated with the occurence of herpes zoster than with varicella infection (p = 0.02). The data generated provide useful background information in our population in the prevaricella vaccine era. PMID- 11422817 TI - Short-term pediatric renal transplant survival: blood pressure and allograft function. AB - Hypertension is prevalent after renal transplantation (Tx) and associated with graft failure in children and adults. However, the effect of blood pressure (BP) on short-term renal allograft function is uncertain. We assessed the associations among BP pretransplant, and 3 months and 1 yr post-transplant, and 1-yr post transplant measured glomerular filtration rate (mGFR) in 61 children with a functioning graft. The GFR was determined using a single intravenous (i.v.) injection of Optiray 350(R). Data were collected between January 1994 and January 2000. The mean mGFR 1 yr after renal transplant was 63.6 +/- 19.9 mL/min/1.73 m2 in 26 live donor recipients and 50.8 +/- 23.3 mL/min/1.73 m2 in 35 cadaveric donors (p = 0.029). Correlation analysis showed significant negative associations of 1-yr mGFR with systolic blood pressure (SBP) and diastolic blood pressure (DBP) 3 months after renal Tx (r = - 0.58, p < 0.0001 and r = - 0.50, p < 0.0001, respectively), and with SBP (r = - 0.37, p = 0.003) and DBP (r = - 0.32, p = 0.01) 1 yr after renal Tx. Multi-variate regression analysis showed that the SBP 3 months after Tx (p < 0.001), number of acute rejections (p = 0.002), donor age (p = 0.02), and cold ischemia time (p = 0.03) were independent predictors for the 1-yr mGFR. These results indicate that a higher SBP in the first few months post renal Tx is associated with decreased renal allograft function in children 1 yr post-Tx. PMID- 11422818 TI - Renin angiotensin system gene polymorphisms in pediatric renal transplant recipients. AB - Genetic variability in the renin angiotensin system may modify renal responses to injury and disease progression. We therefore examined whether the insertion/deletion polymorphism of the angiotensin-converting enzyme (ACE) gene, the Met235-->Thr polymorphism of the angiotensinogen (AGT) gene, and the A1166- >C polymorphism of the angiotensin II type 1 receptor gene (ATR1) were associated with disease progression and outcome after renal transplantation (Tx) in 100 Caucasian pediatric renal transplant recipients. The observed allele frequencies were: DD 31%, DI 41% and II 28%, for ACE; MM 42%, MT 37% and TT 21%, for the methionine (Met)235-->threonine (Thr) polymorphism of AGT; and AA 51%, AC 38% and CC 11%, for the adenine (A)1166-->cytosine (C) gene polymorphism. The slope of 1/creatinine was determined by linear regression analysis of a median of 12 points before and after renal Tx, and the population was divided in two equal groups, according to the slope, both before and after Tx. There were no statistically significant differences for AGT, ACE, and ATR1 polymorphisms with regard to the slope of 1/creatinine before renal Tx. After renal Tx, the ACE II genotype (p = 0.024, chi-square test) and the presence of the I allele (p=0.033, chi-square test) were associated with a favorable slope of 1/creatinine. There was no association of the AGT or the ATR1 polymorphism with outcome after renal Tx, and none of the genotypes were associated with hypertension before or after renal Tx. We suggest that the beneficial association of disease progression after renal Tx with the II genotype and/or the presence of the I allele in our pediatric cohort might be explained by a lower activity of the circulating ACE enzyme associated with the I allele. PMID- 11422819 TI - Comparison of polyclonal induction agents in pediatric renal transplantation. AB - Collective pediatric data suggest that anti-T-cell induction therapy with polyclonal antibodies improves the outcome of both short- and long-term renal allograft survival. Polyclonal agents, including thymoglobulin (Thy), a rabbit anti-thymocyte globulin; Minnesota (horse) anti-lymphoblast globulin (ALG); and ATGAM, a horse anti-thymocyte globulin (ATG), all suppress B and T cells. While no specific T-cell subset marker exists to measure the adequacy of immunosuppression with polyclonal induction, flow cytometric analysis has been used to evaluate the suppression of CD3, CD4, and CD8 cells. Thy is currently undergoing pediatric trials at our center, and we have utilized ATG and ALG in previous pediatric induction protocols. ALG (20 mg/kg/day) and ATG (15 mg/kg/day) were administered over 10 days, whereas Thy (2 mg/kg/day) was given over 5 days. All inductions were accompanied by preoperative intravenous solumedrol (10 mg/kg) followed by oral prednisone (2 mg/kg/day) with taper. Preoperative (1.5 mg/kg/day) and post-operative (2 mg/kg/day) azathioprine was administered to patients receiving ALG or ATG. Mycophenolate mofetil (MMF) (1200 mg/m2/day) was given to the patients receiving Thy. Post-operative cyclosporin A (CsA) (14 mg/kg/day) was started (for all groups) once renal function permitted (creatinine < 50% of baseline with brisk urine output) (trough goal 150-250 ng/mL via HPLC). Values for CD3, CD4, and CD8 T cells were determined by flow cytometry in 2-18-yr old renal transplant recipients, comparing the polyclonal induction agent utilized [Thy (n = 8), mean age 9.7 +/- 2.3 yr; ATG (n = 13), mean age 10.1 +/- 4.1 yr; and ALG (n = 9), mean age 9.3 +/- 3.7 yr] over days 2-10 post-induction. Data were expressed as the average percentage of cells remaining relative to the baseline T-cell subsets (day 1 = 100%), because of the large age variation present in basal T-cell subset values. The flow cytometric data suggest that 5 days of Thy appears to give an equal or greater peripheral blood T-cell suppression by day 10 than a 10-day course of either ATG or ALG. PMID- 11422820 TI - Renal retransplantation of children: is a policy 'first cadaver donor, then live donor' an acceptable option? AB - Retransplantation is often a necessity for children with end-stage renal disease (ESRD), as kidney graft survival is still not infinite. If a suitable live donor is present, the current policy is to use the live donor first, in order to obtain excellent long-term outcome and to prevent human leucocyte antigen (HLA) sensitization. Data from the Eurotransplant International Foundation were analyzed to determine whether the sequence, first a cadaveric donor then a live donor, is acceptable. Between January 1 1983 and December 31 1995, 1305 children received a first renal transplant; 269 of them had a second transplant during the same period. Follow-up of at least 1 yr was available. Categories were made according to the sequence of renal donor source: 217 patients were classified as first cadaver and second cadaver (1cad-2cad) transplant, 26 as first cadaver and second live (1cad-2liv) donor transplant, 23 as first live donor and second cadaver (1liv-2cad) transplant and three patients had two subsequent live donor transplants (1liv-2liv). When a live donor transplant was carried out, either first or second, the donor age was always higher, and the chance of a pre-emptive transplantation or short stay on dialysis was higher, compared with a cadaver transplant. The re-graft survival rate of the '1cad-2liv' was better than the '1cad-2cad' and '1liv-2cad' transplants. At 5 yr, the survival was 76%, 49%, and 61%, respectively. These data suggest that, when a suitable live donor is not available for a first transplantation owing to medical and/or familial reservations, a policy of 'first a cadaver donor then a live donor' transplantation is a viable option and should even be promoted. The pre-emptive stage of the second transplant, probably with a live donor, is additionally advantageous. PMID- 11422821 TI - Echocardiographic findings of hypertrophic cardiomyopathy in children after orthotopic liver transplantation. AB - This study was carried out to compare echocardiographic findings of children taking tacrolimus and cyclosporin A (CsA) after orthotopic liver transplantation (OLT). Echocardiograms of 19 children were reviewed during hospitalizations after OLT, and echocardiograms were performed on 23 children who returned to the clinic for a routine follow-up visit after OLT. Measurements were made of the left ventricle (LV) end-diastolic dimension, and of the thickness of the LV free wall (LVFW) and the inter-ventricular septum (IVS). From these measurements, the LV mass was calculated. LV outflow gradient was measured by using Doppler interrogation. Comparisons were made between patients on CsA and patients on tacrolimus. Children with hypertrophic cardiomyopathy (HCM) were identified. Two patients from the in-patient tacrolimus group were found to have HCM. These two patients had asymmetric septal hypertrophy with dynamic LV outflow obstruction and were successfully treated with propranolol, with or without discontinuing tacrolimus. In the out-patient studies, there was no difference in LVFW and IVS thickness, or LV mass index, between children on CsA and children on tacrolimus. Hence, tacrolimus is associated with the development of HCM in children. The effect of tacrolimus on HCM development may be acute and temporary. More data are needed to determine the incidence of HCM in children on tacrolimus therapy and to establish guidelines for clinicians who follow-up these children. PMID- 11422822 TI - Development of liver size and perfusion after reduced-size liver transplantation in children. AB - The technique of segmental liver transplantation (s-LTx) provides a method to overcome the shortage of suitable livers for small recipients. Patient survival rates are parallel to those obtained with whole liver transplantation (w-LTx). For long-term rehabilitation, adaptive liver growth and adequate perfusion is crucial; however, morphometric and hemodynamic parameters in growing children with s-LTx are not available. Seventeen children who received a s-LTx and 25 with a w-LTx who had follow-up evaluation 1 and 2 yr after LTx were studied. Mean age at time of transplantation was 4.3 +/- 3.5 yr for s-LTx and 10.3 +/- 6.0 yr for w LTx, mean height 98 +/- 21 cm and 122 +/- 30 cm respectively. At follow-up evaluation mean values for liver enzymes, bilirubin and prothrombin time were in the normal ranges for both groups. Liver dimensions were measured by gray scale ultrasound, and hemodynamic parameters by Doppler sonography in the portal vein and hepatic artery using an Acuson 128 machine. Maximal (Vmax), minimal (Vmin) and time-average velocity (TAV) were measured and the resistive index (RI) calculated. We found that 1 and 2 yr after LTx liver dimensions were at a mean in the upper normal range of healthy controls. Spleen size was above the normal range and did not show any tendency towards regression. Mean Vmax in the hepatic artery in s-LTx and w-LTx was 48 cm/sec vs. 28 cm/sec after 1 yr and 30 cm/sec vs. 35 cm/sec after 2 yr, the RI 0.66 vs. 0.55 and 0.59 vs. 0.73, respectively (p for all parameters > 0.05). Maximal portal vein flow was 25 cm/sec in s-LTx vs. 29 cm/sec in w-LTx. Blood flow calculated by vessel diameter and TAV showed no statistical difference between both groups. In conclusion, liver size after s-LTx and w-LTx was increased to the upper normal range, and portal vein blood flow velocities were within the normal range. Vmax in the hepatic artery was reduced in s-LTx; however, the reduction was to the same extent as in w-LTx. In the view of long-term functional adaptation, s-LTx is not inferior to w-LTx. PMID- 11422823 TI - Epstein-Barr virus-related post-transplant lymphoproliferative disease in solid organ transplant recipients, 1988-97: a Canadian multi-centre experience. AB - The aim of this work was to obtain information on the magnitude of the problem, disease characteristics, and clinical practices relating to post-transplant lymphoproliferative disease (PTLD) in Canadian institutions. Adult and pediatric Canadian solid organ transplant groups were sent a questionnaire between July and October 1998. Analyzable data were obtained from 33 transplant groups. For the period 1988-97, 90 cases of PTLD were seen among 4283 solid organ transplant recipients. The incidence of PTLD varied from 0 to 14.6%, with the highest rates in children. Lymph nodes were the sites most frequently affected. Among the classifiable lesions, the majority were monoclonal. The lesions were of B-cell origin in 42.2% and of T-cell in 15.6%. The lesions were classified as monomorphic in 31.1%, polymorphic 18.9%, and hyperplastic in 1.1%. Tumors were reported as low grade in 26.7% and high grade in 10%. The majority of patients (71.1%) received reduced immunosuppression. Anti-viral agents were used in 52.2%. Chemotherapy was used in 27.8%, while immune globulin was used in 22.2%. Surgical resection was used in 20.0%, radiotherapy in 14.4%, and interferon-alpha therapy in 12.2%. The results showed that 48.9% of the patients had died, while 25.6% and 8.9% were regarded as having complete remission and partial remission, respectively. In conclusion, the incidence of PTLD varies widely across Canadian centres. Children are disproportionately affected and the mortality rate is high. Management practices vary significantly, and the need for information sharing was identified as one way of optimizing management. PMID- 11422824 TI - Oral administration of tacrolimus in the presence of jejunostomy after liver transplantation. AB - The feasibility of oral administration of tacrolimus in the presence of an intestinal stoma after liver transplantation (LTx) has not been adequately demonstrated. A 10-month-old girl underwent LTx with biliary reconstruction using a Roux-en Y loop. She developed intestinal perforation and underwent a jejunostomy at 40-50 cm distal to the jejunojejunostomy of the Roux-en Y loop on day 8 post-LTx. Tacrolimus was given twice daily via a nasogastric tube or orally; the initial dose of tacrolimus was 0.10 mg/kg/day. Until the time of intestinal perforation, the trough level of tacrolimus ranged from 13.0 to 19.6 ng/mL. The dose-normalized trough concentration (DNTC) of tacrolimus ranged from 130 to 196 ng.kg.day per mg.mL (control: 80-145 ng.kg.day per mg.mL). For a 2 week period when the patient was septic, the tacrolimus dose was reduced to 0.05 mg/kg/day, with a subsequent trough level of 3.6-5.1 ng/mL (DNTC: 72-102 ng.kg.day per mg.mL). After 3 weeks, the dose was increased to 0.175 mg/kg/day with the disappearance of infection; the trough level ranged from 8.5 to 9.7 ng/mL with a peak level of 26.3 ng/mL (DNTC: 48.5-55.4 ng.kg.day per mg.mL). After the initiation of oral feeding, the dose was slightly increased to 0.20 mg/kg/day with the trough level ranging from 8.1 to 9.8 ng/mL (DNTC: 40.5-49 ng.kg.day per mg.mL). After closure of the jejunostomy, the dose of tacrolimus was reduced to 0.075 mg/kg/day to maintain the same trough level (7.9-9.1 ng/mL) and the DNTC ranged from 105 to 121 ng.kg.day per mg.mL. In conclusion, oral administration of tacrolimus may achieve the therapeutic level, even in the presence of jejunostomy after LTx, although the bioavailability is decreased. PMID- 11422825 TI - Transjugular intrahepatic portosystemic shunt prior to renal transplantation in a child with autosomal-recessive polycystic kidney disease and portal hypertension: A case report. AB - Autosomal-recessive polycystic kidney disease (ARPKD) can cause renal failure and portal hypertension in children. Portal hypertension may complicate the course of renal transplantation (Tx). We report the successful outcome of a patient with end-stage renal disease (ESRD) and portal hypertension treated with transjugular intrahepatic portosystemic shunt (TIPS), a minimally invasive endovascular technique of portosystemic shunt, prior to renal Tx. PMID- 11422826 TI - The 12th Annual Report of the North American Pediatric Renal Transplant Cooperative Study: renal transplantation from 1987 through 1998. AB - The North American Pediatric Renal Transplant Cooperative Study (NAPRTCS) presents an annual report for all transplants registered from January 1987 onwards. In this report we reviewed 6,534 renal transplants recorded for 5,958 patients who had entered the study by January 1999, and attempted to identify changes in practice patterns that had led to improved graft survival. There has been a steady decline in cadaver source transplants nationally and our accrual for 1996 and 1997 reflected this trend. There has also been a decrease in the number of infants and young children receiving a transplant in recent years. From a peak of 23.3% in the 1987-91 cohort, the current report shows that children under 6 yr of age now account for only 20.4% of all transplants. Changing disease patterns and rates of progression of disease have decreased the percentage of Caucasian children in the transplant registry, from 68.5% in the first 5 yr to 62.9% in the most recent cohort. Changing practice patterns have markedly reduced the use of cadaver donor (CD) kidneys (recovered from donors younger than 10 yr of age) from 35% in 1991 to 22% in the current report. Acute rejection patterns are identical for CD and living donor (LD) grafts for the first 2 weeks post transplant. The comparative percentages on days 30 and 45 are 36% and 44% for CD, and 26% and 32% for LD recipients respectively. By the end of the first year post transplant, 45% of LD and 60% CD recipients have had an acute rejection. There has been a marked improvement in our ability to reverse the initial episode of rejection; in 1987, 52% were completely reversed in LD recipients, and in 1997 61% were reversed. Rejection percentages continued to be lower in patients maintained on cyclosporin A (CsA) doses of > 6.4 mg/kg. One-, 3-, and 5-yr graft survival probabilities were 91%, 85%, and 80%, respectively, for LD recipients, and 83%, 73%, and 65% for CD recipients. Comparative 1- and 3-yr figures from 1987 to 1991 were 88% and 81% for LD and 74% and 63% for CD recipients. When short-term graft survival (1 yr) results were compared, a significant improvement was demonstrated from 71.7% in 1987/88 to 92.6% in 1998/1999 for CD transplants. Hence, changing practice patterns have gradually brought the short-term graft survival of CD transplants very close to that of LD transplants. The continuing decrease in the incidence of acute rejections in more recent years should translate into a further delay in the onset of chronic rejection, thus improving graft longevity. PMID- 11422827 TI - A new method to analyze basic rest-activity cycle. AB - A new method was proposed to analyze a basic rest-activity cycle (BRAC), termed BRAC-A. This method calculates a significant cycle length by chi-squared method and the starting points of a resting phase and an active one by analysis of the array consisting of the segmented time series in the same length. Using 20 3-day computer-simulated time series of standard deviation in fetal heart rate fluctuation with known periodicity of the BRAC, we demonstrated that the BRAC-A was a useful tool to reveal the characteristics of the BRAC. PMID- 11422828 TI - Epileptic seizures and sleep-wake rhythm. AB - The subjects were 25 children, including West syndrome, Lennox-Gastaut syndrome, childhood absence epilepsy (CAE) and localization-related epilepsies. Clinical seizures occurred only during waking state in 14 patients (including the cases of West syndrome, CAE and epilepsy with myoclonic absences (EMA)), only during sleeping state (especially during non-REM sleep in five patients, including the case of benign epilepsy with centrotemporal spike), and diffusely during waking and sleeping states in six patients (including the case of early infantile epileptic encephalopathy with suppression burst). These three types were observed in the cases of Lennox-Gastaut syndrome and localization-related epilepsies. Subclinical ictal discharges occurred during REM sleep in West syndrome, and during REM and non-REM sleep especially during non-REM sleep in CAE and EMA. PMID- 11422829 TI - Effects of short nap and exercise on elderly people having difficulty in sleeping. AB - The purpose of the present study was to examine the effects of short nap and exercise on the sleep quality of elderly people who reported difficulty in sleeping. 'Interventions' such as short nap after lunch and moderate-intensity exercise in the evening were carried out for 4 weeks. After the 'interventions', wake time after sleep onset significantly decreased and sleep efficiency significantly increased, which showed that sleep quality was improved. The frequency of nodding in the evening significantly decreased. These results demonstrated that the proper awakening maintenance in the evening was effective in improving sleep quality. After the 'intervention', mental health was also improved with improving sleep quality. PMID- 11422830 TI - Changes in fractal structure of heart rate variability during a nap in one case. AB - The objective of the present study was to analyze fractal structures of adult heart rate (HR) variability during a nap. Fractal analysis was carried out in one case over consecutive 10-min time series of HR, which were simultaneously recorded with electroencephalogram. Scaling relationships showed cross-over patterns characterized by alphas and alphal (i.e. slopes above and below a cross over point). The alphas and alphal were black and white noise at Stage 4 of NREM sleep, and black and 1/f noise in REM sleep. Cross-over points changed from the first to second sleep cycle. We demonstrate the multifractal structures of HR variability during a nap in the present case. PMID- 11422831 TI - Is the sleep initiating process affected by psychological factors? AB - The aim of this study was to investigate the effects of psychological factors on sleep initiating process. The Y-G personality test (Y-G), Lazarus-type stress coping inventory (SCI), Zung self-rating depression scale, General health questionnaire, and Sleep health questionnaire were administered to 418 subjects (mean age = 21.13 +/- 8.76 years), and the data were analyzed by multiple regression analysis (stepwise variable reduction method). Self-rating depression scale; Co-Em, Pla, Sel scale in SCI; C, N, O, Co scale in Y-G were significantly included in the final model. The results showed that psychological factors, such as depression, emotion-focused coping behaviour, prudent planning to solve problems, low self-control, small changes of feeling, nervousness, subjectiveness, and cooperativeness are associated with difficulty initiating sleep. PMID- 11422832 TI - The effects of recreational underwater exercise in early evening on sleep for physically untrained male subjects. AB - The present study investigated the effects of recreational underwater exercise (UWE; 28.9-31.5 degrees C, 16:00-16:50, 36.2 +/- 5.2 %HRres) on thermal response and nocturnal sleep. Rectal temperature (Trec) and sleep parameters of eight physically untrained male subjects were measured in both non-exercise (NE) and UWE days. Underwater exercise consisted of aerobic exercises with various kinds of movements. During UWE, Trec raised slightly in most subjects, but declined in one subject. We could not observe a rapid decline in Trec before sleep on a UWE day. The polysomnographic and subjective sleep parameters showed no significant differences between NE and UWE nights. The quality of sleep did not improve on the night when UWE was performed, and thermal changes might be related to the results. PMID- 11422833 TI - Spatiotemporal changes of slow wave activities before and after 14 Hz/12 Hz sleep spindles during stage 2 sleep. AB - The present study examined the spatiotemporal changes of slow wave (delta and theta bands) activities before and after 14 Hz/12 Hz sleep spindles during stage 2 sleep, using topographic mapping of electroencephalogram (EEG) power. Both types of sleep spindles appeared after slow wave activities of background EEG decreased. Moreover, the appearance of sleep spindles provided increasing EEG slow wave activities in the subsequent period. Further, the present results showed that an appearance of 14 Hz sleep spindle facilitated slow wave activities at the centro-parietal areas, while an appearance of 12 Hz sleep spindle facilitated slow wave activities at the fronto-central areas. These results suggest that sleep spindles provide cortical de-arousal, and serve to maintain sleep. PMID- 11422834 TI - Sleep patterns during 30-m nitrox simulated saturation dives. AB - The sleep patterns were examined during the simulated 30-m nitrox saturation dives. The standard polysomnography of 15 divers was recorded for a total of 255 nights, as were patterns of change or consistency in sleep variables. A reduction of total sleep time in accordance with the lengthening of sleep latency and the wake after sleep onset was observed through the latter part of the bottom period to the post-dive period, but the other sleep variables did not show any changes. These findings suggest that decompression and the psychological stress due to being in the closed environment of a hyperbaric chamber for a long time have effects on divers' sleep. PMID- 11422835 TI - Activation of visual cortex in REM sleep measured by 24-channel NIRS imaging. AB - To visualize dreaming brain functions we studied hemodynamic changes in the visual cortex during the transition from non-rapid eye movement (NREM) to rapid eye movement (REM) sleep, using a 24-channel Near-Infrared Spectroscopy (NIRS) imaging method. Results were compared to the activation in visual cortex by visual stimulation during wakefulness. Subjects were four healthy males between 25 and 49 years of age. Five all-night polysomnographic and NIRS recordings were made. Increases in the oxygenated hemoglobin concentration in visual cortex were observed from nine of 14 REM periods. The activated areas were broader during REM sleep than during visual stimulation. These findings suggest that activation of visual cortex in REM sleep might represent dream-related brain activity. PMID- 11422836 TI - Synchronization between hippocampal theta waves and PGO waves during REM sleep. AB - We investigated the temporal relationship between hippocampal theta waves (referred to as 'theta waves') and ponto-geniculo-occipital (PGO) waves in cats. We measured the time intervals between an occurrence of a PGO wave and the prior and posterior adjacent positive peaks of the theta wave. The unimodal distributions of these intervals suggested a close temporal relationship between PGO and theta waves (i.e. synchronization). In addition, the period of a theta wave during which a PGO wave occurred was not statistically different from that without a PGO wave. This result suggests that the synchronization found here is characterized by phase-locking rather than phase-resetting. Common mechanisms are suggested to underlie the generation of PGO and theta waves. PMID- 11422837 TI - Quantitative characteristics of alpha and theta EEG activities during sensory deprivation. AB - Our previous study showed that theta and alpha electroencephalographic (EEG) activities occurred together during sensory deprivations (SD). The respective activities are expected to manifest different physiological states. In this study, we investigate statistical properties of both types of EEG activities during SD. Wavelet analyses of the original EEG activities during SD and the corresponding surrogation data can distinguish meaningful alpha and theta activities in the original data from accidentally detected ones. As a result, the mean duration of the theta wave is found to be longer than that of the alpha wave. This result could be useful to disclose physiological processes during SD. PMID- 11422838 TI - Spatio-temporal EEG power spectral patterns during a short daytime nap. AB - This is an approach to investigate topographic changes in electroencephalographic (EEG) spectral power during pre- and post-nap wakefulness as well as stages 1 (S1) and 2 (S2) NREM sleep in 12 subjects. Delta- and theta-band power significantly increased in the frontal and central regions during S1 and S2 with an increase in inter- and intra-hemispheric correlations. Beta-band power significantly increased in the frontal, central and parietal regions during S2 with an increase in interhemispheric correlation. In contrast, alpha-band power significantly decreased in the parietal-occipital regions during S1 and S2 with a decrease in interhemispheric correlation. Thus, daytime nap modulated spatio temporal patterns of EEG power spectral patterns in wide scalp regions. PMID- 11422839 TI - Two auto-detection methods for eye movements during eyes closed. AB - Eye movements during closed eyes closely reflect changes of the arousal level during transition from wakefulness to sleep. Because they contain both rapid and slow eye movements (REM and SEM), it has been difficult to detect them automatically. Hiroshige recently developed the method of linear regression analysis for automatic detection of the two types of eye movements, and we have developed a template matching method for autodetection. The aim of the present study was to compare both auto-detection methods and visual scoring for REM and SEM. The results revealed high agreement between the two quantitative methods and the visual scoring, indicating that auto-detection of eye movements is useful for quantitative evaluation of arousal level. PMID- 11422840 TI - Examination of accuracy of sleep stages by means of an automatic sleep analysis system 'Sleep Ukiha'. AB - We examined the differences between the results of an automatic sleep analysis system and inspection decision. Subjects were 10 males (average age 21.6 years). One section consists of 20 s records. The sections that deviated from the algorithm could not be decided. Each sleep stage decided by automatic analysis was compared with the inspection decision. The agreement ratio of stage 3 was 91.6% in the highest, and followed by stage 2, stage 4, stage W and stage 1. The lowest was 62.5% for movement time. The total agreement ratio was 85.8%. The agreement ratios of the apnea index (AI) and the apnea hypopnea index (AHI) were relatively high, but for types of sleep apnea, agreement ratios require improvement. PMID- 11422841 TI - Age and gender difference in habitual sleep-wake rhythm. AB - We analysed the effects of age and gender on the habitual sleep-wake rhythm in 2252 randomly selected subjects living in Shimonoseki, Japan. Subjects were divided into 21 age groups with a matching number of males and females for each. During the period from primary school to adolescence, the bedtime became delayed and sleep length decreased with age. After that period, with increasing age, bedtime became earlier and sleep length increased. The number of awakenings and the length of daytime naps increased markedly with epoch after 50 years and 70 years of age, respectively. Gender differences were considered to be due to the women's social and domestic customs in Japan. PMID- 11422842 TI - Narcolepsy and other non-SAS hypersomnia in sleep breathing disorders clinic. AB - Four of the 708 snorers (0.56%), referred to our sleep breathing disorders clinic for the past 2 years were diagnosed as having narcolepsy-cataplexy. Detecting HLA DRB1*1501/DQB1*0602 positive was informative for differentiating genuine narcolepsy from non-sleep apnea syndrome (non-SAS) hypersomnia in our clinic. A non-SAS obese boy, diagnosed as having essential hypersomnia syndrome, was found to be HLA DRB1*1502/DQB1*0601 positive. His hypocretin concentration was 206 pg/mL in the cerebrospinal fluid. PMID- 11422843 TI - Characteristics of patients with hypnotic-related psychiatric disorders in the nationwide mental hospital survey. AB - We investigated the characteristics of patients with hypnotic-related psychiatric disorders. Subjects were 90 patients reported in the nationwide Mental Hospital Survey in 1998, who were divided into three groups according to their history of substances use. Group A (patients without history of any illicit substance use) was estimated to be the most common and core group with iatrogenic characteristics. Hypnotic use in Group B (patients with history of any illicit substance use) was recognized as one of behavioral disorders in the psychosocial context. Patients in Group C (patients with multiple-substance use and without history of any illicit substance use) showed the most serious disorders in the context of addiction. More specified treatment programs would be required considering patients' characteristics. PMID- 11422844 TI - Study on withdrawal of hypnotics: questionnaire on hypnotic use and its withdrawal. AB - To investigate the situation and problems contingent to hypnotic use and withdrawal, we conducted a questionnaire of outpatients. Only 41% of the patients were satisfied with their sleep and 53% of the patients took hypnotics. As regards the period, 83% of users had used them for more than 1 year and 19% had used them for more than 10 years. Although 90% of patients perceived efficacy of hypnotics, 67% felt more or less anxious about hypnotic use. Sixty-seven per cent of patients had actually withdrawn from the drugs or decreased dosage before. More than half the patients' conditions worsened after the withdrawal or reducing dosage. PMID- 11422845 TI - A survey of junior high school students' sleep habit and lifestyle in Okinawa. AB - A survey was made of the sleep and lifestyle activity patterns of 3754 students from 14 different junior high schools on Okinawa Island. The survey showed that bedtimes became progressively and significantly later as students ascended to higher grades, resulting in adolescent sleep debt. The later adolescents retired to sleep, there appeared significantly greater numbers suffering from insufficient sleep, who found difficulty in waking up, who arose later in the mornings, and who failed to eat breakfast. The study indicated that along with later bedtimes, staying up late resulted in deterioration of sleep health, poor dietary habits and feeling ill, contributing to sleepiness during class. PMID- 11422846 TI - Subjective sleep quality and sleep problems in the general Japanese adult population. AB - To dimensionally describe subjective sleep quality using the Pittsburgh Sleep Quality Index (PSQI) and estimate the prevalence rate of sleep problems (PSQI global score > 5) in the general Japanese adult population, 1871 subjects randomly selected from the 1995 Census were examined. The PSQI component scores (mean +/- SD) widely ranged (e.g. 0.04 +/- 0.31 in hypnotic medication use for males aged 20-29 years, 0.03 +/- 0.18 in hypnotic medication use for females aged 20-29 years, 1.10 +/- 0.94 in sleep latency for males aged 80 or older, 1.52 +/- 1.03 in sleep latency for females aged 80 or older). Statistical significance was found in each component score among age groups by gender. The PSQI global scores (mean +/- SD) by age groups ranged from 4.00 +/- 2.59 to 5.02 +/- 3.89 for males (P < 0.39) and 4.30 +/- 2.34 to 6.75 +/- 4.10 for females (P < 0.001). The respective prevalence rates of sleep problems were 26.4% (95% CI = 23.6, 29.3) for males and 31.1% (95% CI = 28.1, 33.9) for females. PMID- 11422847 TI - Estimated sleep debt and work stress in Japanese white-collar workers. AB - Sleep debt on weekdays in Japanese white-collar workers, estimated using a questionnaire, was associated with age, overtime, and self-rated workload. PMID- 11422848 TI - Light conditions during sleep period and sleep-related lifestyle in Japanese students. AB - The effects of light conditions during night sleep on sleep habits and morning evening preference were studied in Japanese students (18-28 years old). Students who usually used fluorescent or non-fluorescent light on the room ceiling, wall or desk preferred to have a daytime nap significantly later (Mean: 14:50 h) than those who used no light (13:34 h). Students who used no curtain or a half transparent lace-curtain showed shorter sleep latency (duration from going-to-bed to sleep-onset) than those who used a curtain which shuts off lights from outside. Light conditions during the middle of night and early in the morning may affect the timing of sleep in Japanese students based on the circadian system. PMID- 11422849 TI - Seasonal changes in sleep and behavioral problems in a pubescent case with autism. AB - This study reports on the seasonal changes in sleep problems and behavioral problems in a pubescent case with autism. His care diary, kept for 1 year, showed seasonal changes of sleep and behavioral problems, and that his behavioral problems preceded his sleep problems. PMID- 11422850 TI - A longitudinal study of diurnal variation in baseline fetal heart rate in one dichorionic-diamniotic twin pregnancy. AB - A longitudinal study to analyse the diurnal variations in baseline fetal heart rate (FHR) and sustained fetal tachycardia (SFT) in twin pregnancy was performed on one dichorionic twin. Twenty-four hour FHR recordings on twins were made at 32, 34 and 36 weeks of gestation simultaneously. Significant diurnal variations were found in both twins in all gestational weeks. The diurnal variations in baseline FHR of twins were highly correlated with no phase-lag. No coincidence was recognized in the appearance of SFT between twins. These results suggest the maternal influence equally affects FHR in each fetus of dichorionic twin since 32 weeks of gestation, while the appearance of SFT might be independent from maternal influence. PMID- 11422851 TI - Comparisons of sleep patterns between mothers in post-partum from 9 to 12 weeks and non-pregnant women. AB - In order to evaluate two patterns of interrupted and non-interrupted sleep for the post-partum mothers from 9 to 12 weeks after delivery, we compared them with sleep patterns of non-pregnant women. Subjects were 10 primipara and 12 non pregnant women. Their polysomnographic recordings were made using a Medilog recorder at home. In interrupted sleep, low sleep efficiency, decreased total sleep time, and a decreased percentage of stage 2 were significantly observed compared with non-pregnant women. Sleep parameters of non-interrupted sleep, except for increased percentage of stage 4, did not show any significant differences from non-pregnant women. Mothers' sleep fluctuated between interrupted sleep similar to the early post-partum sleep from 1 to 6 weeks and non-interrupted sleep with increased stage 4. PMID- 11422852 TI - Development of body temperature rhythm: 6 years follow up of three cases. AB - The present study was conducted to evaluate longitudinally the relationship between phase of temperature rhythm and sleep habits. Sleep-logs were recorded from three children for 3 weeks during school term and 1 week before the experiment, and oral temperature was recorded for 2 consecutive days during summer vacations for 6 years. Although previous studies have reported that phase of temperature rhythm was established by 5 years old, phase delay of temperature was found in all subjects even after 5 years old. These results suggest that the establishment of temperature phase is after 5 years old. PMID- 11422853 TI - Age-related changes of sleeping pattern during adolescence. AB - A questionnaire concerning sleep-related lifestyle, and physical and mental health was administered to the students of junior high schools (12-15 years), high schools (15-18 years), and universities. While bedtime linearly delayed with age throughout the adolescence (from junior high school to university age), rise time remained constant until high school ages. As a result, total nocturnal sleep time considerably decreased during this period. Rise time then delayed with age in the university sample. Daytime sleepiness increased with age peaking at high school age, then decreased gradually during university age. Several physical and mental health conditions got worse during high school and university ages. Compared with the adolescent samples, daytime sleepiness and mental health complaints were very low in the adult samples. PMID- 11422854 TI - Ethanol effect on sleep electroencephalogram by the correlation dimension. AB - The influence on sleep of alcohol (ethanol) was investigated in nine males using the correlation dimension (CD). Polysomnography (PSG) was recorded on a baseline night (BL-N) and on an ethanol (0.8 g/kg) night (Et-N). The mean correlation dimensions on Et-N, as well as on BL-N, significantly decreased from sleep stage 'awake' to sleep Stages 1, 2 and 3 and increased during rapid eye movement (REM) sleep. The mean correlation dimensions of the sleep EEG during sleep Stage 2 and those for the second sleep cycle on the Et-N were significantly higher than those on the BL-N. The changes in the correlation dimensions between sleep cycles were reduced on Et-N as compared to BL-N. PMID- 11422855 TI - The effects of flumazenil on sleepiness, task performance and nocturnal sleep after anesthesia with midazolam. AB - The purpose of the present study was to clarify the changes in psychophysiological functions after anesthesia with midazolam (intravenous (i.v.) benzodiazepine anesthetic) and to examine the ability of flumazenil (benzodiazepine antagonist) to prevent the adverse effects of anesthesia with midazolam. Clinical dose of midazolam (0.1 mg/kg i.v.) was administered to seven healthy volunteers and either flumazenil (0.3 mg i.v.) or saline was injected at the end of the anesthesia. After anesthesia with midazolam, subjective sleepiness and euphoria increased significantly, but these changes were not observed when flumazenil was administered. In addition, sleep latency was prolonged and sleep efficiency decreased significantly after midazolam anesthesia with and without flumazenil. Slow wave sleep decreased significantly only by co-administration of flumazenil. PMID- 11422856 TI - Effects of isoflurane and ketamine on sleep in rabbits. AB - The postoperative sleep disturbance (POSD) is characterized by reduction of sleep after surgical operation. However, its mechanism is not well known. Therefore, we hypothesized that anesthetics could contribute to the POSD, and studied the effects of isoflurane and ketamine on sleep in rabbits. Rabbit sleep was measured for 21 h after isoflurane exposure or intravenous injection of ketamine. Non rapid eye movement sleep (NREMS) was decreased after isoflurane anesthesia. In contrast, ketamine anesthesia significantly enhanced NREMS. Both anesthetics did not affect rapid eye movement sleep. These results suggest that isoflurane may contribute to the POSD, but ketamine may decrease the POSD. PMID- 11422857 TI - Comparison in symptoms between aged and younger patients with narcolepsy. AB - We investigated the age-related changes in symptoms in narcolepsy. Fifty patients, 65-year-old and over (aged group), were recruited from the National Narcolepsy Registry. Thirty-four patients, younger than 65 (younger group), were selected by random sampling. Although there was no difference in the age of disease onset between the two groups, the age of diagnosis was significantly earlier for the younger group. Methylphenidate was used significantly more in the aged group, and modafinil in the younger group. The aged group had lower total scores on the Ullanlinna Narcolepsy Scale, because the scores for cataplexy were significantly less for the aged group. There was no significant difference in excessive daytime sleepiness between the two groups. PMID- 11422858 TI - Narcoleptic canines display periodic leg movements during sleep. AB - Periodic leg movements during sleep (PLMS) is a high prevalent sleep disorder of unknown etiology. The disease is pharmacologically treated with dopaminergic agonists (i.e. D2/D3 agonists) and opiates. Periodic leg movements during sleep often occur in narcoleptic patients. We observed that narcoleptic canines, like narcoleptic humans, also exhibit jerky, unilateral or bilateral slow leg movements during sleep. The movements in dogs are characterized by repetitive dorsiflexions of the ankle, lasting 0.5-1.5 s, and occur at regular intervals of 3-20 s, thus showing similarities to PLMS in humans. The observation that D2/D3 agonists aggravate cataplexy in narcoleptic dogs suggests that altered dopaminergic regulation in canine narcolepsy may play a critical role in both cataplexy and PLMS. Our canines may therefore be an invaluable resource in PLMS research. PMID- 11422859 TI - Long-term follow-up study on patients with sleep apnea syndrome. AB - We did a 5-year follow-up study on sleep apnea syndrome (SAS)-related symptoms and respiratory disorder variables on 109 patients. At follow up, SAS-related symptoms were improved in a significantly higher percentage of the treatment success group than the treatment failure group. A significant deterioration of the respiratory disorder variables was recognized only in the patients 40-60 years of age. These results indicate that treatment effect for SAS might be maintained in the long term and that the natural evolution of the disorder might be prominent in middle-aged patients. PMID- 11422860 TI - P300 abnormalities in patients with severe sleep apnea syndrome. AB - We investigated the mechanism of abnormal P300 findings in the patients with severe sleep apnea syndrome (SAS). Compared with controls, SAS patients showed significantly longer latency of P300. Not the mean sleep latency in multiple sleep latency test but the percentage of total periods in which percentage desaturation time fell below 90% to total sleep time correlated with P300 latency in the patients. During nasal CPAP treatment, P300 latency was significantly shortened in the patients under 45 years, while the elderly patients did not show statistical change. We speculated that P300 latencies might be prolonged due to nocturnal hypoxia with SAS and the abnormality might be irreversible, especially in elderly patients. PMID- 11422861 TI - A trial study of RhinoSleep for the diagnosis of sleep apnea. AB - RhinoSleep was developed to diagnose obstructive sleep apnea (OSA). The RhinoSleep consists of a soft catheter, a measuring probe, and a computer. The soft catheter is inserted through the naris into the esophagus. The catheter is connected to the measuring probe, which creates and sends a continuous noise signal into the catheter. When the catheter is compressed during an obstruction, the sound reflections change, making it possible to identify both the level and the duration of the obstruction. Sixteen OSA patients were examined using RhinoSleep and daytime polysomnography simultaneously. With RhinoSleep, we successfully examined 13 cases. This trial study identified problems with RhinoSleep, such as vomiting of the catheter, hardware instability, and the criteria decision for the analysis of OSA. PMID- 11422862 TI - A comparison of middle latency auditory-evoked response in obstructive sleep apnea syndrome before and after treatment. AB - The ascending reticular activating system seems to be affected by nocturnal hypoxia in obstructive sleep apnea syndrome (OSAS) patients so that their vigilance level is decreased. To evaluate the change in vigilance level in OSAS patients, polysomnography, the Epworth Sleepiness Scale and middle latency auditory-evoked responses (MLR) were measured before and after treatment in seven men with OSAS (mean age 45.4 +/- 12.9 years, mean body mass index 33.3 +/- 7.6 kg/m2). After treatment there was significant improvement of nocturnal hypoxia in OSAS. Peak amplitude of P1 in MLR increased after treatment. There was also improvement of the electrical field distribution of MLR on the scalp. The results suggest that the decreased vigilance level in OSAS patients is reversible with treatment. PMID- 11422863 TI - Aggregated occurrence of sleep apnea syndrome in a family. AB - We experienced a family in whom sleep apnea syndrome (SAS) was recognized in six members and habitual snoring in seven members among 26 subjects of four generations. In all members with snoring, the disorder was noticed before the age of 20. Hypercapnic response study showed normal findings, and ultrafast magnetic resonance image of the upper airway revealed that the obstruction of the upper airway occurred at the velopharyngeal portion during apneic episodes in all SAS afflicted subjects. In this family, the similar craniofacial structural abnormalities were thought to cause the aggregated occurrences of the disorders at young ages, and obesity was regarded as a contributor for the aggravated symptoms. PMID- 11422864 TI - Association of gastroesophageal reflux disease with weight gain and apnea, and their disturbance on sleep. AB - Obesity is a common predisposition to gastroesophageal reflux disease (GERD) and obstructive sleep apnea syndrome (OSAS). By statistical analysis of the respondents to a questionnaire that was distributed to members of the Kansai Rugby Association, we examined whether weight gain increased the incidence of these diseases and whether GERD alone disturbs sleep. Prevalence distribution of GERD by age differed from another survey, which suggests that predispositions other than age may contribute to GERD. Weight gain tended to increase the incidence of GERD. In our epidemiological study, both GERD (particularly nocturnal reflux) and OSAS significantly contributed to sleep disturbance. Although GERD alone seemed to be one of several independent factors of sleep disturbance, it was not a weak factor. PMID- 11422865 TI - Effects of physiological cardiac pacing on sleep-disordered breathing in patients with chronic bradydysrhythmias. AB - In six patients with chronic bradydysrhythmias, polysomnographies were performed before cardiac pacemaker implantation and over the week following implantation. A patient with third-degree atrioventricular block (AVB) and two patients with sinus node dysfunction (SND) were associated with sleep-disordered breathing (SDB). Their cardiac pacemaker therapies, with the increase in the average heart rate, led to a reduction of apnea-hypopnea index and/or an improvement of Cheyne Stokes breathing. It seems that chronic bradydysrhythmia is one of the causative factors leading to SDB. PMID- 11422866 TI - Comparison of Ricketts analysis and Downs-Northwestern analysis for the evaluation of obstructive sleep apnea cephalograms. AB - To determine which analysis is suitable to examine the dentofacial skeletal pattern characteristics of the obstructive sleep apnea syndrome (OSAS), we took lateral cephalograms of 44 Japanese OSAS patients and 34 Japanese non-OSAS controls. By Ricketts analysis, we found significant differences between OSAS patients and non-OSAS controls on facial axis, lower facial height and total facial height, which showed that Japanese OSAS patients have dolico facial patterns. However, by Downs-Northwestern analysis, we did not find any significant difference between OSAS patients and non-OSAS controls using the same cephalograms as the Ricketts analysis. PMID- 11422867 TI - Uvulopalatopharyngoplasty for obstructive sleep-related breathing disorders: one year follow up. AB - The effectiveness of uvulopalatopharyngoplasty was evaluated in 28 patients (25 men and three women; mean age 47.6 years) with obstructive sleep apnea by comparing preoperative daytime polysomnography with those carried out 6 months and 1 year postoperatively. The mean apnea plus hypopnea index (AHI) in the supine position decreased from 51.0/h to 23.0/h 6 months postoperatively and 57.1% of patients showed a 50% or more reduction of AHI. By changing the sleep position from supine to lateral, 71.4% of the 28 patients indicated a 50% or more reduction in AHI before surgery, which increased to 92.3% of 26 patients 6 months postoperatively. The positive results of uvulopalatopharyngoplasty were maintained at least until 1 year after surgery. PMID- 11422868 TI - Amyotrophic lateral sclerosis associated with insomnia and the aggravation of sleep-disordered breathing. AB - A case of amyotrophic lateral sclerosis (ALS) diagnosed by sleep-disordered breathing is described. The patient's chief complaints were insomnia and nocturnal dyspnea after taking a hypnotic drug. On examination, he showed restrictive ventilatory impairment, alveolar hypoventilation and hypoxia. Polysomnographic examination revealed marked hypoxia during REM sleep periods, decreased duration of REM sleep periods, and increased sleep disruption. Amyotrophic lateral sclerosis was diagnosed by the neurological finding of paraspinal muscle weakness and neurogenic changes revealed by needle electromyography and muscle biopsy. The daytime and nocturnal respiratory insufficiency improved after nasal bilevel positive airway pressure therapy. Amyotrophic lateral sclerosis should be suspected as a cause of insomnia and nocturnal dyspnea. PMID- 11422869 TI - Questionnaire relating to sleep paralysis. AB - A sleep survey was conducted on 8162 citizens. The cumulative experience rate of sleep paralysis was 39.6%. The initial occurrence of sleep paralysis peaked at age 16 years. In addition to being higher in young people than in older subjects, the incidence of sleep paralysis was also higher among women than among men, and was significantly higher among shift worker than non-shift worker, and among persons engaged in the nursing profession than those not engaged in the nursing profession. The experience rate of sleep paralysis demonstrated a strong correlation with the frequency of dreaming, the experience rate of nightmares, times and regularity of going to bed and waking up, and particularly with the degree of insomnia. PMID- 11422870 TI - Melatonin therapy for REM sleep behavior disorder. AB - Rapid eye movement sleep behavior disorder (RBD) is a parasomnia with clinical symptoms that include punching, kicking, yelling and leaping out of bed in sleep. Polysomnographic (PSG) finding showed REM sleep without muscle atonia. Clonazepam is generally used for treating RBD symptoms but melatonin was reported to be effective so we reconfirmed the effect of melatonin on RBD patients in the present study. We used melatonin (3-9 mg/day) which could ameliorate problem sleep behaviors remarkably, as well as %tonic activity in PSG variables. In the present study, melatonin was reconfirmed to be effective in RBD symptoms, especially for patients with low melatonin secretion, while its mechanism was not clearly known in the present study. PMID- 11422871 TI - Basal forebrain microinjections of L-3,3',5-triiodothyronine modify sleep in hypothyroid rats. AB - Male rats were injected bilaterally with various doses of L-3,3',5 triiodothyronine (T3) into basal forebrain areas. The electroencephalogram (EEG), electromyogram (EMG) and brain temperature (Tbr) were then measured in 8-h studies. In the medial preoptic area (MPA), a 4 microg dose of T3 caused significant elevations in REM sleep as compared to control injections. In the median preoptic nucleus (MnPO), both 2 and 4 microg doses of T3 significantly inhibited non-REM sleep. Injections to the diagonal band of Broca did not alter EEG-defined sleep. Influences on Tbr were not significant for any of the treatments. Since these effects of T3 were demonstrated after acute injections, the data are consistent with possible non-genomic actions of thyroid hormones in adult brain. PMID- 11422872 TI - Parasomnia as an occasion for the diagnosis of Parkinson's disease. AB - We report a case of Parkinson's disease (PD) diagnosed by REM sleep behavior disorder (RBD). The patient was a 68-year-old man. On admission, rigidity in the left upper and lower extremities, bradykinesia, and gait disturbance were noted. In addition, polysomnography revealed REM sleep without atonia (RWA), and a diagnosis of untreated PD associated with RBD was made. Polysomnographic data showed that REM density decreased and RWA tended to increase after administration of a combination of L-DOPA and DCI (L-DOPA/DCI). Thus, we considered that the pathophysiological mechanism of RBD in this case was based not only on the dysfunction of the brainstem mechanism of RWA, but also on the impairment of dopaminergic neuron. PMID- 11422873 TI - Re-entrainment of the circadian rhythms of plasma melatonin in an 11-h eastward bound flight. AB - We investigated the re-entrainment of melatonin rhythm in an 11-h eastward-bound flight. Eight male subjects participated in the present study. Blood sampling was carried out once before the flight and twice after the flight. During the daytime the subjects were exposed to natural zeitgeber outdoors on the day except the blood sampling. Seven of eight subjects showed antidromic re-entrainment, and the other subject showed orthodromic re-entrainment. The intensity of natural day light in New York amounted to 20 000 lx. As for the direction of the re entrainment in New York the antidromic re-entrainment is naturally dominant. PMID- 11422874 TI - Nineteen-hour variation of postural sway, alertness and rectal temperature during sleep deprivation. AB - Nineteen-hour variation of postural sway, alertness and rectal temperature during sleep deprivation were studied. Alertness decreased gradually at night and remained low until morning. Postural sway in the eyes-closed condition increased during early morning. In six of the eight subjects the greatest sway was observed during the 3-h period when rectal temperature was at its minimum. It is suggested that unbearable sleepiness during sleep deprivation will give rise to measurable impairment of postural balance especially during the time zone of temperature nadir. PMID- 11422875 TI - Sleep-wake rhythm during stay in an intensive care unit: a week's long-term recording of skin potentials. AB - To monitor the sleep-wake cycle of patients during their stay in the intensive care unit (ICU), we tried continuous and long-term recording of skin potential (SP) levels in patients after surgery. A graph of the week-long SP showed the sleep-wake pattern to be evident until the fourth day. It disappeared beginning on the fifth day, resulting finally in delirium with a relatively high mean SP level. In another record, the administration of sedative agents to calm the excitement lowered the mean SP level and suppressed SP responses evoked by frequent day and night treatment or nursing care. Continuous monitoring of arousal level by SP will be of help in prevention of ICU syndrome. PMID- 11422876 TI - Effects of vitamin B12 on bright light on cognitive and sleep-wake rhythm in Alzheimer-type dementia. AB - The present study investigated the effects of vitamin B12 (VB12) on circadian rhythm in Alzheimer-type dementia (ATD). Twenty-eight ATD patients were treated with bright light therapy (BLT) for 8 weeks. For the latter 4 weeks, half were treated with VB12 with BLT (BLT + VB12). We evaluated the cognitive state with Mini-Mental State Examination and the circadian rhythm with actigraphy after the fourth and eighth week. After the first 4 weeks BLT improved the circadian rhythm disturbances and cognitive state especially in the early stage of ATD. Although the latter 4 week-BLT caused no significant effects on the circadian rhythm; BLT + VB12 improved the vigilance level during the daytime. These results suggest that VB12 has some efficiency to enhance vigilance for ATD patients. PMID- 11422877 TI - Effects of bright light exposure on heart rate variability during sleep in young women. AB - To investigate the effects of evening bright light on the autonomic nervous system, heart rate variability (HRV) during sleep was analyzed in dim light (DL) and bright light (BL) conditions. We recorded polysomnography in nine healthy young women aged 20-21 years. Time series of % delta power was calculated in the 0.49-2.20 Hz band. Heart rate variability was analyzed from a 10-min segment of slow wave sleep. The low- to high-frequency ratio and the low-frequency component decreased significantly in the BL conditions compared with the DL conditions. However, the power of the high-frequency component did not change in the two conditions. These results indicate that evening BL affects the autonomic nervous system during slow wave sleep. PMID- 11422878 TI - Effects of light and sleep stages on heart rate variability in humans. AB - Effects of light intensity and sleep stages on heart rate variability (HRV) were investigated in young healthy subjects. The low-frequency (LF)/high-frequency (HF) ratio was significantly increased by exposing either to bright lights of 10 000 lx or to extreme darkness (< 0.01 lx), while HF and LF components of HRV were not changed, when compared with those under dim light (100 lx). However, LF was significantly increased at REM sleep, when compared with that at the pre-sleep wake. In contrast, HF was increased at all stages of sleep, and the LF/HF ratio was decreased at slow wave sleep during the baseline night. PMID- 11422879 TI - Effects of bright light at lunchtime on sleep of patients in a geriatric hospital I. AB - The effects of lunchtime bright light exposure in patients of a geriatric hospital were investigated. Ten inpatients (six women and four men; mean age +/- SD: 81.2 +/- 8.8 years) with sleep disturbances were studied for 9 weeks. Nurses performed daily ratings for sleep-wakefulness disturbances. Approximately 8000 lx bright light exposure was performed for 3 weeks in the light therapy room. Before and after exposure, ocular function was evaluated. Clinical ratings of sleep wakefulness improved in eight patients. The score of difficulty in falling asleep and drowsiness in the morning declined during the light exposure. The score of drowsiness in the afternoon decreased during the post-light exposure. Post exposure ocular disturbances were not found. PMID- 11422880 TI - Effects of bright light at lunchtime on sleep in patients in a geriatric hospital II. AB - Inpatients with sleep disturbances in a geriatric hospital received 1 h of exposure to approximately 8000 lx bright light per day for 3 weeks. Polysomnogram was recorded for four female patients. Two (aged 68 and 87 years) were non demented patients with weak cataracts and the other two (aged 92 and 93 years) were demented patients with severe cataracts. Electroencephalogram results showed that light exposure decreased the proportion of Stage W, while increased the proportion of Stage 2, and these effects continued for at least 3 weeks after the cessation of light exposure. These results suggest that exposure to bright light is effective in improving the disturbed sleep of patients. PMID- 11422881 TI - A circadian system model with feedback of cross-correlation between sleep-wake rhythm and oscillator. AB - Recent findings suggest that social entrainment factors work on an oscillator other than the temperature and melatonin rhythms. This indicates that there is a feedback loop affected on oscillators. In the present study, a circadian system model, including mutually coupled two oscillators and sleep-wake rhythm, is constructed, where cross-correlation between a sleep-wake rhythm and an oscillator is set to modify the coupling from one oscillator to the other. Based on the model simulation, a mechanism underlying delayed sleep phase syndrome is elucidated by the reduction in the cross-correlation resulting from dissociation between the delayed sleep phase and the oscillator. PMID- 11422882 TI - The effects of oral melatonin on the autonomic function in healthy subjects. AB - We investigated the influence of melatonin on the human autonomic functions by measuring muscle sympathetic nerve activity (MSNA). Five healthy male volunteers took 3 mg of melatonin, and their plasma melatonin concentration, blood pressure, heart rate, and burst rate of MSNA were then recorded. The peak level of melatonin concentration showed a marked interindividual variation. Blood pressure was reduced significantly, while heart rate and burst rate of MSNA did not change significantly. These results indicate that melatonin has a hypotensive effect on blood pressure, and the central cardiovascular regulatory mechanism such as lowering of the baroreflex setpoint would be involved in the effect. PMID- 11422883 TI - Effect of melatonin on sleep and rectal temperature of young healthy evening types. AB - The present study assessed in healthy nocturnal juveniles the presence or absence of the hypnotic action of exogenous melatonin and its effect on core temperature by oral administration of exogenous melatonin at a pharmacologically low dose 15 min before going to bed at the normal time. Both sleep latency and the number of instances of waking up after falling asleep tended to decrease, while the amount of Stage 2 sleep increased significantly. However, no hypothermic action regarding the rectal temperature for changes in the circadian phase were observed. According to those results, it is possible that exogenous melatonin has some direct hypnotic action on habitual night sleep, although the extent of this action is considered to be relatively weak. PMID- 11422884 TI - Hypnotic action of melatonin during daytime administration and its comparison with triazolam. AB - The present study was conducted to assess hypnotic action, effects on rectal temperature and dose dependency by daytime administration of exogenous melatonin (MLT) at 1 mg, 3 mg or 6 mg to subjects consisting of seven healthy juvenile adults. As a result, exogenous MLT significantly increased total sleep time and sleep efficiency, and MLT 6 mg was observed to demonstrate hypnotic effects that were nearly equal to those of triazolam at 0.125 mg. Rectal temperature was significantly decreased at MLT 1mg and 3 mg, there were no significant differences observed in the hypothermic effects at MLT 6 mg. These results indicate that exogenous MLT had dose-dependent hypnotic action on daytime sleep, and it is possible to consider that this hypnotic action was based on a direct acting mechanism. PMID- 11422885 TI - Proposed supplements and amendments to 'A Manual of Standardized Terminology, Techniques and Scoring System for Sleep Stages of Human Subjects', the Rechtschaffen & Kales (1968) standard. PMID- 11422886 TI - Subcutaneous emphysema associated with chest tube drainage. AB - BACKGROUND: Subcutaneous emphysema may complicate a pneumothorax, but may also occur as a consequence of its treatment by chest tube drainage. The aim of this study was to evaluate the factors involved in the association between subcutaneous emphysema and chest tube drainage, and the clinical outcomes in these cases. METHODOLOGY: One hundred and sixty-seven patients undergoing chest tube drainage within a 12-month period were evaluated retrospectively. There were 30 reported cases of subcutaneous emphysema (SE). Comparisons were made between those with subcutaneous emphysema and those who did not develop this complication. RESULTS: A total of 134 patient notes were evaluated (25 SE and 109 non-SE). Subcutaneous emphysema was more commonly associated with trauma, bronchopleural fistulae, large and bilateral pneumothoraces, and mechanical ventilation. Subcutaneous emphysema was also associated with prolonged drainage, poor tube placement, tube blockage, side-port migration, and a greater number of chest tubes. Importantly, those with SE had a longer length of stay and increased mortality. CONCLUSION: Subcutaneous emphysema can be spontaneous or traumatic, but is associated with avoidable causes such as inadequate chest tube drainage, particularly due to poor tube placement, anchorage and blockage, and also with side-port migration into the subcutaneous tissue. It is associated with an increased morbidity and mortality, and may indicate the need for urgent chest tube replacement. PMID- 11422887 TI - Cell type accuracy of transthoracic fine needle aspiration material in primary lung cancer. AB - OBJECTIVE: The aim of this study was to evaluate the diagnostic accuracy of transthoracic fine needle aspiration (TFNA) materials in establishing the specific cell type in primary lung cancer, and to study the influence of several factors on this accuracy. METHODOLOGY: The present study included 129 patients [(12 females, 117 males; mean age 54.6 years (range 25-75)] who underwent thoracotomy. The initial diagnosis was obtained by means of TFNA biopsy in all patients. Transthoracic fine needle aspiration was performed by 22-gauge Chiba needle with fluoroscopy guide in 93 patients and with computed tomography guide in 36 cases. RESULTS: The overall concordance was 73.6% (Kappa = 0.52). The worst agreement was obtained for the large cell carcinoma (40%; Kappa = 0.48). The likelihood of a correct diagnosis using the TFNA specimens was 6.2-fold higher for well-differentiated tumours than for poorly differentiated tumours (P < 0.005). The stage of tumour and diameter of the lesion had no effect on cell agreement. Cell agreement was higher in central lesions than peripheral lesions, but the difference was not statistically significant (P = 0.097). This difference was more significant between patients with central and peripheral epidermoid carcinoma (P = 0.057). CONCLUSION: In our opinion, cell typing by TFNA may lead to incorrect results in the presence of poor differentiation, mixed tumours and peripheral epidermoid carcinomas. PMID- 11422888 TI - Inspiratory muscle performance in endurance athletes and sedentary subjects. AB - OBJECTIVE: The aim of this study was to determine whether whole-body endurance training is associated with increased respiratory muscle strength and endurance. METHODOLOGY: Respiratory muscle strength (maximum inspiratory pressure (PImax)) and endurance (progressive threshold loading of the inspiratory muscles) were measured in six marathon runners and six sedentary subjects. RESULTS: PImax was similar between the two groups of subjects but the maximum threshold pressure achieved was greater in marathon runners (90 +/- 8 vs 78 +/- 10% of PImax, respectively, mean +/- SD, P < 0.05). During progressive threshold loading, marathon runners breathed with lower frequency, higher tidal volume, and longer inspiratory and expiratory time. At maximum threshold pressure, marathon runners had lower arterial O2 saturation, but perceived effort (Borg scale) was maximal in both groups. Efficiency of the respiratory muscles was similar in both groups being 2.0 +/- 1.7% and 2.3 +/- 1.8% for marathon runners and sedentary subjects, respectively. CONCLUSIONS: The apparent increase in respiratory muscle endurance of athletes was a consequence of a difference in the breathing pattern adopted during loaded breathing rather than respiratory muscle strength or efficiency. This implies that sensory rather than respiratory muscle conditioning may be an important mechanism by which whole-body endurance is increased. PMID- 11422889 TI - The relationships between patients' related variables in asthma: iplications for asthma management. AB - OBJECTIVE: Understanding patients' personal characteristics is essential for better asthma management. This study assessed the relationships between patients' related variables in asthma and identified key associations relevant to asthma management. METHODOLOGY: Subjects were recruited from the Alfred Hospital Asthma and Allergy Clinic (Melbourne, Victoria, Australia) and general practices. Their clinical and demographic characteristics, asthma knowledge, impact of asthma on their quality of life, their self-management skills and attitudes to asthma were assessed. RESULTS: One hundred and sixty-nine subjects participated in the study. Fifty-seven per cent had one or more previous hospital admissions, 94% had either moderate or severe asthma, and 51% reported nocturnal symptoms in the last 6 weeks. Patients who spoke only English, had been admitted to an intensive care unit, had a peak flow meter, and an asthma action plan had significantly better asthma knowledge than those who did not. The impact of asthma was greatest in patients who had a peak flow meter, used oral steroids, had exercise limitation, and developed asthma between the ages of 31-45 years. Female patients had better self-management skills than males. Patients with asthma-related distress were more likely to use oral steroids or theophylline and to have a history of previous hospital admissions. Patient self-confidence was negatively correlated with age. CONCLUSIONS: There are significant relationships between many of patients' variables related to asthma, including their personal clinical, demographic and psychological characteristics. The findings have implications for optimizing asthma management. PMID- 11422890 TI - Tachykinin-induced bronchoconstriction in sheep is NK-1 receptor mediated and exhibits tachyphylaxis. AB - OBJECTIVE: Tachykinins are mediators of airway hyper-reactivity and inflammation. There is in vitro evidence that ovine responses to tachykinins correlate closely to human responses. This study was designed to characterize the effect of intravenously administered tachykinins on sheep lung resistance in vivo to determine the effect of dose timing on reproducibility of responses and the induction of tachyphylaxis. We then used this information to help further characterize the response with several pharmacological agents. METHODOLOGY: Substance P (SP) was administered by infusion to conscious merino ewes and lung resistance (RL) was measured. Infusions were given at 30, 60, 120 min and 24 h intervals. The effect of various agents on the response to SP was then assessed. RESULTS: Substance P led to a transient increase in RL, mean (+/- SEM) 754.8 (+/- 139)% of baseline, with marked tachyphylaxis at 30, 60 and 120 min. Phosphoramidon increased the peak response to 1151.5 +/- 196%. Atropine and CP 96 345 abolished the response to SP, while indomethacin, sodium cromoglycate and pyrilamine had no significant effect. Substance P had a greater effect on RL than did neurokinin A. CONCLUSIONS: Substance P increases RL in sheep via a cholinergic mechanism which is mediated by NK-1 receptors, and is subject to tachyphylaxis. These findings have implications for the design of studies using the ovine model in the evaluation of tachykinin antagonists as potential therapeutic agents. PMID- 11422891 TI - Assessment of factors predicting outcome of acute respiratory distress syndrome in North India. AB - OBJECTIVES: Information on the spectrum and outcome of acute respiratory distress syndrome (ARDS) in tropical countries is scanty. This study was designed to assess the factors predicting the outcome of ARDS in North India. METHODOLOGY: Consecutive patients requiring mechanical ventilation for ARDS over a 2 year period at the Respiratory Intensive Care Unit (RICU) of a tertiary referral hospital were studied. Hospital survival was correlated with age, aetiology, disease severity scores (APACHE III, SAPS II, lung injury score) and organ failure using univariate analysis. Factors significantly influencing mortality were examined by multivariate analysis to identify factors independently affecting outcome. RESULTS: Sepsis (28.6%), followed by malaria (21.4%), were the commonest risk factors. Seven out of eight patients (87.5%) with sepsis died. The presence of sepsis, more than three organ failure prior to admission, APACHE III score > 57 and SAPS II score > 39 were significantly associated with mortality. Only APACHE III score > 57 or SAPS II score > 39 were, however, independently predictive of a poor outcome following multivariate analysis. CONCLUSIONS: Sepsis, associated with a very poor outcome, and malaria were important risk factors for the development of ARDS. APACHE III score > 57 or SAPS II score > 39 were associated with increased risk of mortality. PMID- 11422892 TI - Effect of the long-term use of inhaled corticosteroids on bone mineral density in asthmatic women. AB - OBJECTIVE: Inhaled corticosteroids have become a key element in the maintenance treatment of bronchial asthma. Recent studies have shown that administration of inhaled corticosteroids is associated with evidence of derangement in bone turnover. Therefore, we studied the bone mineral density (BMD) of asthmatic women receiving long-term inhaled corticosteroids and compared them with healthy individuals matched for age, sex, menopausal status and body mass index. METHODOLOGY: Thirty-two female patients with bronchial asthma, who had been using inhaled corticosteroids (beclomethasone dipropionate 750-1500 microg/day) regularly for at least 3 months, were included in the study. Bone mineral density measurements were done with dual X-ray absorptiometry in the lumbar area of the spine and the hip. Detailed laboratory examination was also done for the patients and 26 controls. RESULTS: There was a significant decrease in BMD of the patient group at the lumbar region and femur as compared with normal controls. In the patients there was a significant negative correlation between the duration of therapy, daily and cumulative doses, and BMD at the lumbar region but not BMD at the femur. CONCLUSIONS: These results indicate that long-term use of inhaled corticosteroids is associated with significant bone loss in asthmatic women and is especially related to the duration of therapy. Therefore, it is necessary to appropriately screen and give prophylactic treatment to those who are likely to develop osteoporosis from inhaled corticosteroid treatment. PMID- 11422893 TI - Replication-deficient adenovirus-mediated transfer of B7-1 (CD80) cDNA induces anti-tumour immunity in isolated human lung cancer. AB - OBJECTIVE: Interaction between the co-stimulatory molecule B7-1 (CD80) on antigen presenting cells and its counter-receptor CD28 on T lymphocytes plays a key role in the induction of cell-mediated immune responses. Many tumours, including lung cancer, lack expression of B7-1 and this has been suggested to contribute to the failure of immune recognition of these diseases. Based on this knowledge, we hypothesized that the co-stimulatory signal delivered through the B7-1 molecule expressed on lung cancer cells using replication deficient adenovirus vector would induce efficient tumour immunity in T lymphocytes. METHODOLOGY: To evaluate this hypothesis, we constructed two adenovirus vectors: AdCMVhB7 (an E1-deleted Ad5-based vector containing human B7-1 cDNA driven by cytomegalovirus immediate early promoter and enhancer) and AdNull (same vector as above without expression of exogenous gene) as control. Using these adenovirus vectors, efficient generation of tumour immunity in T lymphocytes was studied using primary cultured lung cancer cells and peripheral blood lymphocytes (PBL) obtained from patients with lung cancer. RESULTS: Inoculation of lung cancer cells with 10 multiplicity of infection of AdCMVhB7 resulted in rapid and efficient cell surface expression of B7-1 molecule (> 90% of cells at 24 h). Cytolytic activity of PBL in 51Cr release assay (E/T = 40) demonstrated that effector lymphocytes induced by hB7-1 (+) lung cancer cells treated with AdCMVhB7could lyse parental lung cancer cells hB7-1 (-). In contrast, effector lymphocytes induced by lung cancer cells treated with AdNull as control virus or PBS as control could not lyse parental lung cancer cells at all. Furthermore, cytolytic activity of the effector lymphocytes induced by B7-1-transduced lung cancer cells was inhibited by addition of anti CD3 antibody. CONCLUSIONS: These data demonstrate that primary-cultured lung cancer cells treated with AdCMVhB7 would efficiently generate tumour immunity in T lymphocytes. Adenovirus-mediated-hB7-1 gene transfer may be a useful means for gene therapy of lung cancer using adoptive immunotherapy. PMID- 11422894 TI - Early unplanned readmission of patients with newly diagnosed tuberculosis discharged from acute hospital to ambulatory treatment. AB - OBJECTIVE: The aim of this study was to examine whether patients with newly diagnosed tuberculosis (TB) discharged to ambulatory treatment are at risk of unplanned readmission through the emergency department within 28 days of discharge, and the risk factors associated with such readmission. METHODOLOGY: A cohort of 134 patients admitted to an acute medical department with TB, who were subsequently discharged to ambulatory treatment of TB, were studied by a retrospective record review for unplanned readmission in 28 days. Potential risk factors associated with the readmission were recorded during hospital stay and follow-up visits, including age, sex, length of stay, substance abuse, need of assistance in the activities of daily living (ADL), comorbidities, non compliance, drug complications and use of non-standard drug regimen. RESULTS: Up to 20.1% of patients were readmitted. Factors independently associated with early unplanned readmission were need of assistance in ADL, drug complications, the need to use a non-standard drug regimen and more than three non-chest comorbidities. CONCLUSIONS: A significant readmission rate was found in these patients and potential risk factors were identified. Ambulatory treatment for TB may not be appropriate for selected patients. Local guidelines for the management of TB patients at high risk of readmission is needed. PMID- 11422895 TI - Exercise-induced bronchoconstriction among Malay schoolchildren. AB - OBJECTIVE: Exercise-induced bronchoconstriction (EIB) may be demonstrated in 60 70% of asthmatic children in temperate climates. In areas of high humidity it is postulated to be low. The aim of the study was to determine the prevalence of EIB in a population of schoolchildren with wheezing, living in the humid tropical climate of Kuala Lumpur, Malaysia. METHOD: We performed a cross-sectional study using the International Study of Asthma and Allergies in Childhood questionnaire to identify 7-12-year-old Malay children with asthma symptoms from a primary school in central Kuala Lumpur. Sixty-five of 76 children with 'ever wheeze' performed an exercise challenge test successfully in an uncontrolled environment. A random sample of 80 schoolchildren with no history of wheeze were similarly tested as controls. The relative humidity and temperature were recorded. A fall of > 15% was considered as clinically important. RESULTS: The prevalence of EIB in schoolchildren with 'ever wheeze' was 47.7%. The prevalence of EIB in children with 'current wheeze' was 51.6%. The prevalence of EIB in controls was 7.5%. The relative humidity during the study ranged from 41 to 90%. There was no significant relationship between different humidity levels and EIB (P = 0.58, regression analysis). CONCLUSION: This study demonstrates that EIB is present in asthmatic children despite the highly humid tropical environment. PMID- 11422896 TI - Craniofacial abnormalities in Japanese patients with severe obstructive sleep apnoea syndrome. AB - OBJECTIVE: To clarify that factors besides obesity play an important role in the development of obstructive sleep apnoea syndrome (OSAS) in Japanese patients, we compared craniofacial structures in patients with severe OSAS with those of normal controls. METHODOLOGY: The craniofacial structures of 60 Japanese patients with severe OSAS and 30 normal controls were evaluated using standard cephalometric analysis. Patients were stratified according to body mass index (BMI): non-obese, BMI < 25; moderately obese, BMI = 25-30, severely obese, BMI > 30. RESULTS: The SNA (sella to nasion to subspinale angle) was significantly smaller in the patient groups than in the controls. The SNB (sella to nasion to supramentale angle) and NSBa (cranial base flexure) were significantly smaller in the non-obese and moderately obese patients than in controls. The MP-H (distance from the mandibular plane to the hyoid bone) and the PNS-P (distance from the posterior nasal spine to the tip of the soft palate) were significantly longer in the patient groups than in the controls. The PNS-P was significantly longer in the severely obese patients than in the non-obese patients. CONCLUSIONS: Japanese patients with severe OSAS have enlargement of the soft tissues and palate as well as craniofacial bony structural abnormalities. This is particularly apparent in non-obese patients. PMID- 11422897 TI - Hypoxic ventilatory depression in a patient with mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes. AB - We describe a case of a 21-year-old man with mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS) who presented with hypoxic ventilatory depression. He had chronic hypoventilation, which was not explained by weakness of respiratory muscles. His hypercapnic ventilatory response was not impaired. In contrast, hypoxic ventilatory depression was observed in the isocapnic progressive hypoxic response test. After exposure to hypoxic conditions, his respiratory frequency decreased and tidal volume was unchanged. The hypoxic ventilatory depression was partially blocked by pretreatment with aminophylline. In conclusion, we need to be careful with patients with MELAS who are hypoxaemic because a vicious circle of hypoxia and hypoventilation can occur. PMID- 11422898 TI - Hepatopulmonary syndrome associated with autoimmune liver cirrhosis. AB - A 46-year-old woman presented for evaluation of liver dysfunction and dyspnoea. Laboratory examination showed high levels of gamma-globulin, immunoglobulin (Ig)G, and antinuclear antibodies. Laparoscopy demonstrated hepatic cirrhosis. Despite normal spirometry, hypoxaemia (which was worse in standing position) and a low diffusing capacity were present. The shunt ratio calculated using arterial blood gas was 6.4%, but was 40% when measured using 99mTc-macroaggregated albumin scanning. The discrepancy between the ratios indicated that hypoxaemia was caused by intrapulmonary vascular dilatation. The patient was diagnosed with hepatopulmonary syndrome associated with autoimmune liver cirrhosis. PMID- 11422899 TI - Lemierre's syndrome. AB - This is a case report of Lemierre's syndrome in a 17-year-old male patient. Lemierre's disease consists of suppurative thrombophlebitis of the internal jugular vein (SIJVT) in the presence of oropharyngeal infection and can be complicated by septic pulmonary embolism. Other causes of SIJVT include deep neck infections and central venous catheterization. The disease usually results from Gram-negative anaerobic organisms such as Fusobacterium necrophorum. PMID- 11422900 TI - Il-12: keeping cell-mediated immunity alive. AB - Interleukin (IL)-12 has dominated the field of cell-mediated immunity since its discovery more than 10 years ago, and clearly plays an essential role in the development of Th1 cells under a variety of conditions. Recent studies now indicate that the importance of IL-12 is not limited to initiating an immune response, but may contribute to maintaining immunity. Thus, Th1 responses rapidly wane in the absence of IL-12, leading to a loss in protective immunity against intracellular pathogens, such as Leishmania and Toxoplasma. Determining how IL-12 maintains Th1 cells, and consequently cell-mediated immunity, will provide new insights useful in controlling the immune response, and may thus influence the design of new vaccines and immunotherapies. PMID- 11422901 TI - Regulation of IL-18 expression in virus infection. AB - Since its discovery as an interferon (IFN)-gamma-inducing factor, it has become evident that interleukin (IL)-18 plays a crucial role in the generation of protective immunity against microbial infections. Macrophages are the major source of biologically active IL-18, and they express constitutively IL-18 mRNA and proIL-18 protein. Microbial infections enhance the IL-18 gene expression in macrophages but post-translational processing of proIL-18, rather than transcriptional activation, is predominant in the regulation of IL-18 secretion. This review summarizes the current knowledge of proinflammatory and immunomodulatory properties of IL-18, and focuses on the role of caspases in the proteolytic activation of IL-18 in response to virus infection. PMID- 11422902 TI - Molecular modelling and endoplasmic reticulum retention of mutated TCR/CD3 complexes. AB - T cell receptor (TCR)/CD3 complex assembly takes place in the endoplasmic reticulum (ER). Normal TCR/CD3 complexes egress from the ER to the cis-Golgi, where the interaction with zeta2 homodimers occurs. This interaction leads to further uncontrolled transport of TCR/CD3/zeta molecules to the cell surface. The purpose of the present experiments was to determine firstly the basis for the impact of the Phe195/216 --> Val mutations on TCR/CD3 expression in Jurkat cells, and secondly why mutated J79-cell TCRalphabeta/CD3 hexamers are prevented from interacting with zeta2 homodimers. We found that Phe --> Val mutations cause serious perturbations in a so far undefined hydrophobic area formed by the two Phe195/216 on beta-strand F and aromatic/large hydrophobic amino acids on neighboring beta-strands B and A in Calpha and Cbeta domains, respectively. In addition, TCR/CD3 hexamers and zeta2 homodimers colocalize in normal Jurkat T cells, in revertant J79r58 cells, and in J79 cells transfected with wild-type TCRalpha cDNA but not in J79 mutant cells (confocal microscopy). Furthermore, mutated TCR/CD3 complexes seem to be actively retained in the ER in J79 cells but not in revertant J79r58 cells by a nondominant mechanism. We propose that a hitherto undefined ER-retention molecule controls both the protein structure and egress of TCR/CD3 complexes from the ER of alphabeta and gammadelta T cells. PMID- 11422903 TI - Characterization of mouse mammary tumour virus-induced migration of lymphoid cells into lymph nodes. AB - We previously found that Mtv-2+ lymph nodes (LN) implanted into Mtv-2- mice underwent marked hyperplasia owing to the influx of lymphocytes. LN grafts infected with exogenous mouse mammary tumour viruses (MMTV), MMTV(FM) transmitted by FM mice and MMTV-2 produced by Mtv-2, also swelled in MMTV-free recipients. Mtv-3 and Mtv-7 also displayed this capability. Mtv-2-induced LN hyperplasia was earlier in onset and greater in extent when major histocompatibility complex (MHC) class II I-E was expressed than unexpressed. Mtv-3-induced LN hyperplasia was suppressed completely by Mtv-3 from a different mouse strain and partially by Mtv-6 slightly different from Mtv-3 in superantigen (SAg) Vbeta specificity. LN hyperplasia occurred bidirectionally in LN transplantation between mice carrying Mtv-2 and Mtv-3, which are different SAg Vbeta specificity. LN hyperplasia induced by MMTV-2 carrying SAg responsive to Vbeta14 alone and MMTV(FM) carrying SAg responsive to Vbeta14 and Vbeta8.2 was completely but partially suppressed by MMTV(FM) and MMTV-2, respectively. CD4+ T cells were essential for MMTV-induced LN hyperplasia. LN in situ also underwent significant hyperplasia when infected with MMTV. Thus, MMTV SAg may entice circulating lymphocytes into lymphoid organs and contribute to more efficient dissemination MMTV in vivo. Secondary lymphoid tissue chemokine (SLC) may not be directly involved in this event. PMID- 11422904 TI - Recent thymic emigrants (CD4+) continuously migrate through lymphoid organs: within the tissue they alter surface molecule expression. AB - T-cell progenitors migrate from bone marrow (BM) into the thymus. After maturation they are released as recent thymic emigrants (RTE) into the periphery ensuring the diversification of the T-cell repertoire. Both the kinetics with which RTE migrate through the periphery and the surface molecules they express are still unclear. In 1- and 18-month-old Lewis rats CD4+ RTE were identified in blood, spleen, lymph node, and thoracic duct lymph by flow cytometry (CD45RC- and CD90+), were differentiated from CD4+ naive (CD45RC+) and memory T cells (CD45RC CD90-), and were characterized regarding the expression of surface molecules. Both in 1- and 18-month-old animals the percentage of RTE among the CD4+ population in blood was comparable to that in all other compartments. Surprisingly, RTE expressed alpha4-integrin, LFA-1, and interleukin (IL)-2 receptor at a significantly higher level than naive T cells and more comparable to memory T cells. Within lymphoid tissues RTE, naive, and memory T cells significantly upregulated the expression of CD44 and ICAM-1, and downregulated the expression of L-selectin. These changes were reversed before the cells re entered the blood. Thus, our data indicate that CD4+ RTE travel through the periphery of young and old rats like mature T cells, continuously modulating their surface molecule expression. PMID- 11422905 TI - Possible involvement of IL-12 in reovirus type-2-induced diabetes in newborn DBA/1 mice. AB - This study extends our previous observations that the reovirus type-2(Reo-2) can induce autoimmune insulitis, which may be mediated by T-helper (Th) 1-dependent mechanisms, resulting in diabetes in newborn DBA/1 mice. In this study mRNA expression for Th1-related cytokines including Th1 and Th2 cytokines in splenic cells was examined by reverse transcriptase polymerase chain reaction (RT-PCR) in relation to the development of insulitis. Furthermore, the effect of monoclonal antibody (MoAb) against interleukin (IL)-12(p40) on the development of insulitis and the mRNA expression in the splenic cells was examined. The mRNA expression for IL-12(p40), IL-18, and interferon (IFN)-gamma, but not IL-5, increased in the spleen in parallel with the development of insulitis. The treatment with MoAb to IL-12(p40) reduced the insulitis with diabetes which was associated with a decrease in the mRNA expression for IL-12(p40), IL-18 and IFN-gamma, and an increase of IL-4 mRNA expression in the spleen. The present study suggested that Th1-dominant systemic immune responses, being responsible for the development of autoimmune insulitis, might be induced by IL-12-induced and IL-18-activated mechanisms. PMID- 11422906 TI - Maturation of dendritic cells by recombinant human CD40L-trimer leads to a homogeneous cell population with enhanced surface marker expression and increased cytokine production. AB - Dendritic cells (DC) have been shown to be potent inducers of specific cytotoxic T-cell responses both in vivo and in vitro. Furthermore, exposure to cytokines such as tumour necrosis factor (TNF)-alpha or CD40 triggering changes DC phenotype and cytokine production and may enhance the T-cell activating capacity of the DC. We studied DC phenotype and cytokine production as well as the T-cell proliferation and cytotoxic T lympocyte (CTL) activation induced by DC generated in vitro. In addition, the effect of exposure to recombinant human CD40L-trimer (huCD40LT) on these parameters was investigated. Effective differentiation of monocytes derived from freshly isolated peripheral blood mononuclear cells (PBMC) was obtained with granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin (IL)-4. The DC expression of human leucocyte antigen (HLA) molecules, CD80, CD83, and CD86 was markedly enhanced by exposure to huCD40LT even compared to TNF-alpha exposure. Only a moderate cytokine production was observed initially, while TNF-alpha addition or CD40 triggering, especially, induced enhanced production of IL-6 and IL-12 p40. Surprisingly, comparable induction of T-cell proliferation by a DC allostimulus or through the presentation of PPD, and influenza M1-peptide specific CTL activity was obtained with nonmaturated (CD83-) and maturated (CD83+) DC. In conclusion, a final maturation of monocyte-derived DC through huCD40LT resulted in a highly homogeneous cell population with enhanced surface marker expression and high production of pro-inflammatory cytokines. In addition, the induction of responses to allo or recall antigens presented by huCD40LT maturated DC was comparable to the responses obtained with the DC maturated through TNF-alpha exposure. PMID- 11422907 TI - In vitro evaluation of an enhanced human serum amyloid A (SAA2) promoter regulated soluble TNF receptor fusion protein for anti-inflammatory gene therapy. AB - Tumour necrosis factor (TNF)-alpha contributes to the pathogenesis of many inflammatory diseases. Recombinant soluble TNF receptor fusion proteins (sTNFR:Ig) are potent TNF antagonists, both in vitro and in vivo. The concentration of serum amyloid A (SAA) increases by up to 1000-fold during inflammation, largely owing to cytokine-driven transcriptional upregulation. A reporter plasmid, comprising the proximal 0.7 kb of the human SAA2 promoter fused to a luciferase gene, was used in transient transfection experiments in human HepG2 hepatoma cells to assess the quantitative and qualitative TNF antagonist properties of a construct in which sTNFR:Ig synthesis is under the control of a chimera of the SAA2 promoter and a tat/HIV element. The SAA2-tat/HIV-sTNFR:Ig construct retained the fine-tuned cytokine responsiveness of the SAA2 promoter, while exhibiting the quantitatively enhanced level of protein expression conferred by the tat/HIV element. It produced a biologically significant TNF inhibition that was at least as strong as that achieved using a CMV promoter driven sTNFR:Ig construct. There was a dose- and time-dependent relationship between the pro-inflammatory cytokine used, and the generation of TNF antagonist activity by SAA2-tat/HIV-sTNFR:Ig. Although sTNFR:Ig protein can be induced by either TNF-alpha or interleukin (IL)-1beta, its antagonist activity is limited to the former cytokine. The SAA2-tat/HIV-sTNFR:Ig construct, and derivatives thereof, may therefore be ideally suited to gene therapy applications that require the local production of potent and specific immune modifiers only when there is active pathology. It may consequently be of particular use in the future treatment of diseases such as rheumatoid arthritis. PMID- 11422908 TI - The effect of cytokines on the activation-induced apoptosis of B cells in autoimmune NZB x NZW F1 mice. AB - Programmed cell death (apoptosis) is an essential process in the development of various tissues and its involvement has been proposed for the elimination of self reactive immature T and B lymphocytes when self antigens are first encountered. In order to further investigate the role of apoptosis in the pathogenesis of autoimmune disease, the apoptosis of lipopolysaccharide (LPS)-activated B cells, peritoneal cells from NZB x NZW F1 (NZB/W F1) mice and nonautoimmune BALB/c mice were assayed using an in vitro culture system. Splenic B cells were isolated and then stimulated with LPS before further activated with crosslinking antimu antibody. In addition, the apoptosis of peritoneal cells induced by crosslinking antimu antibody was also analyzed. The data revealed that the specific apoptosis of both activated B cells and peritoneal cells induced by crosslinking antimu antibody was very similar comparing NZB/W F1 and nonautoimmune BALB/c mice. This activation-induced B-cells apoptosis could be rescued, however, with the addition of cytokines such as interleukin (IL)-5 or IL-10, to the culture. The results suggest that there is no endogenous defect in the apoptosis of activated B cells for autoimmune NZB/W F1 comparing nonautoimmune BALB/c mice. Notably, however, abnormally high levels of the type 2 T helper (Th2)-related cytokines such as IL 5 or IL-10 may play an important role in the abnormal expansion of activated B cells in autoimmune NZB/W F1 mice. PMID- 11422909 TI - Elevated levels and different repertoire profile of colostral anti-LPS antibodies may have a significant role in compensating newborn immunity. AB - A high prevalence of systemic infections caused by enterobacteria such as Escherichia coli is observed during the neonatal period. Lipopolysaccharide (LPS) is one of the major factors responsible for septic shock caused by these Gram negative bacteria. We have recently demonstrated the presence of anti-LPS immunoglobulin (Ig)G antibodies in cord blood with a repertoire identical to that found in maternal serum. In the present study, we analyzed anti-LPS O111 antibody isotypes in maternal serum and colostrum from mothers and in cord serum from their respective full-term (n = 30) and preterm (n = 13) neonate infants. The main isotype found in serum samples from mothers of term infants was IgM (range between 28 and 54 mg/l), followed by IgA (1-2 mg/l) and IgG (2-3 mg/l). The range of IgG antibody concentrations in cord blood was between 2 and 3 mg/l, as a result of placental transfer. A novel observation in our study was that the LPS bands recognized by colostral antibodies were completely different from those recognized by IgG in serum. Colostral IgA antibodies recognized several bands not bound by serum IgG antibodies from the respective maternal serum, independently of the antibody quantity. In addition, we verified the pattern of LPS recognition by serum IgA and colostral IgA antibodies was identical, what suggested that the antibody isotype found in serum could probably be derived from differentiated IgA positive cells which were homing to the mucosa through the mucosal homing mechanism. Identical pattern of recognition was obtained comparing the IgA and IgM isotypes in colostrum. Slight differences in the pattern of recognition were found between colostral and serum IgM antibodies. The fact that colostral antibodies recognize much more bands than serum antibodies may be important for the host to mount an effective immune response in the intestinal lumen, in order to prevent excessive absorption of LPS, reducing possible systemic effects caused by the molecule. PMID- 11422910 TI - Acanthamoeba polyphaga in rheumatoid arthritis: possibility for a chronic infection. AB - Acanthamoeba polyphaga (AP) is ubiquitous in nature and frequently infects humans. AP has some features, such as persistence, which makes it an attractive candidate in studies of a possible infectious aetiology in rheumatoid arthritis (RA). In this study the occurrence of AP-specific antibodies was compared between RA patients and matched controls. PMID- 11422911 TI - Inherited complete factor I deficiency associated with systemic lupus erythematosus, higher susceptibility to infection and low levels of factor H. AB - Here we describe two new cases of complete deficiency of factor I (fI) in two sisters from a consanguineous Brazilian family. The eldest sibling (20-year-old) developed systemic lupus erythematosus (SLE) early during childhood while the youngest had been committed on several occasions owing to repeated infections although she was asymptomatic for auto-immune diseases. We also detected lower concentrations of C3 and factor B in both sisters. Biological functions dependent on complement activation such as the production of opsonins and killing of phagocytozed micro-organisms, chemotactic factors and haemolytic activity were all significantly reduced in both probands. Consistent with the absence of fI and low levels of fH, a deregulated production of C3b was observed by bidimensional electrophoresis in sera of both the probands. PMID- 11422912 TI - Hereditary human complement C3 deficiency owing to reduced levels of C3 mRNA. AB - An 8-year-old son (L.A.S.) of consanguineous parents, presented recurrent bacterial infections, vasculitis and extremely low levels of serum C3 (0.15 microg/ml). The classical and alternative pathway haemolytic activities and the generation of opsonins and chemotactic factors derived from the activation of the complement system were markedly affected in the proband's serum. An in vitro addition of purified C3 restored the classical pathway-dependent haemolytic activity of his serum. Autoradiographs of the proband's lipopolysaccharide (LPS) stimulated and 35S-labelled fibroblast supernatants after that the SDS-PAGE revealed no C3 alpha or beta chains. The amount of C3 mRNA synthesized by the proband's fibroblasts, as evaluated by reverse transcription-polymerase chain reaction (RT-PCR) assays, was greatly reduced. PMID- 11422913 TI - How important is atheromatous renal artery stenosis as a cause of end-stage renal disease? AB - In recent years atheromatous renal artery stenosis has been proposed as a common and potentially preventable cause of end-stage renal disease (ESRD). Occlusive renal artery disease causes renal failure that can be reversed following successful revascularization, but this scenario is relatively rare. Nonocclusive renal artery stenosis is commonly found in association with varying degrees of renal impairment. However, recent evidence and clinical observation suggest that a cause and effect relationship is unlikely. Experimentally, split renal function studies in cases of unilateral stenosis find similar degrees of renal impairment in both kidneys. Clinically, severe renal impairment is often found in the presence of unilateral stenoses. Another nephropathic process must be going on, probably a combination of hypertensive and atheroembolic damage. It is not surprising that the two pathologies often coexist, in view of their shared causes (principally hypertension and tobacco smoking). Previous studies of the importance of renal artery stenosis in causing ESRD probably included many cases of hypertensive and atheroembolic nephropathy. Unfortunately the potential for renal revascularization to prevent ESRD has been exaggerated. PMID- 11422914 TI - Hormone replacement therapy in postmenopausal women with end-stage renal disease: a review of the issues. AB - Hormone replacement is an integral part of therapies to prevent osteoporosis in postmenopausal women and may be considered a component in the treatment of dyslipidemia, cardiovascular disease, and possibly cognitive function. The indications for, and efficacy and prescription of, hormone replacement therapy in postmenopausal women with ESRD have been infrequently studied and less than 10% of postmenopausal women on dialysis are receiving hormone replacement. Small studies suggest that hormone replacement therapy is valuable in treating the dyslipidemia of women on dialysis, but indicate that a reduction in the dosage of hormone replacement may be needed. A potential role for hormone replacement therapy in the treatment and/or prevention of osteoporosis and sexual dysfunction in postmenopausal women on dialysis exists as well. PMID- 11422915 TI - Protein intake in patients with renal failure: comments on the current NKF-DOQI guidelines for nutrition in chronic renal failure. AB - The National Kidney Foundation Clinical Practice Dialysis Outcomes Quality Initiative (DOQI) guidelines recently recommended dietary protein intake for patients with chronic renal failure as follows: predialysis patients should receive 0.60 g/kg/day of protein and increase intake to 0.75 g/kg/day for subjects who cannot follow such a diet. For stable maintenance hemodialysis patients, the recommended protein intake is 1.2 g/kg/day, and for chronic peritoneal dialysis patients, 1.2-1.3 g/kg/day. We differ with these recommendations and believe that a dietary protein intake of 0.8 g/kg/day is appropriate for the predialysis population; an intake of 0.9-1.0 g/kg/day and 1.0 1.1 g/kg/day for maintenance hemodialysis patients and peritoneal dialysis patients, respectively, should be adequate. The rationale and the evidence supporting our arguments are outlined and discussed. PMID- 11422916 TI - Profit-making in the treatment of chronic kidney disease: truth and consequences. AB - Many have speculated that the quality of dialysis care differs for patients treated in for-profit and not-for-profit facilities. In 1999 we published a study that demonstrated poorer survival and lower rates of listing for renal transplant for patients in for-profit rather than not-for-profit, freestanding centers. While these findings generated substantial discussion, few have commented on their implications. In this article we first discuss potential sources of bias that could impact on the study's results and place the findings in context. We conclude that these disparities are likely to be real, as they are consistent with theorized differences between for-profit and not-for-profit health care organizations and with other evaluations of dialysis facility ownership. We then discuss several policy options for addressing the quality differences we identified. Reducing the outcome discrepancies will not be easy. Upon considering several policy alternatives, we conclude that a widespread effort to link processes of dialysis care to patient outcomes is best suited to reduce the quality differences between for-profit and not-for-profit dialysis units and improve outcomes in both types of facilities. PMID- 11422917 TI - The dialysis outcomes and practice patterns study (DOPPS): how can we improve the care of hemodialysis patients? AB - Outcomes among dialysis patients vary considerably internationally and across regions within the United States. The Dialysis Outcomes and Practice Patterns Study (DOPPS) is a large, prospective, observational study of representative samples of hemodialysis patients in France, Germany, Italy, Japan, Spain, the United Kingdom, and the United States. The DOPPS collects a wealth of data regarding the patients' demographic characteristics, medical histories, laboratory values, prescriptions, dialysis unit practices, and outcomes. The study seeks to clarify which dialysis practices contribute to improved mortality rates, hospitalization rates, health related quality of life, and vascular access outcomes, after adjusting for the effects of comorbid disease and demographic variables. Over 18,000 patients have been enrolled to date. This paper describes the initial findings and outlines the plans to expand the trial. PMID- 11422918 TI - What clinically important advances in understanding and improving dialyzer function have occurred recently? PMID- 11422923 TI - Blood flow surveillance of hemodialysis grafts and the dysfunction hypothesis. AB - It is widely recommended that hemodialysis graft surveillance programs should be implemented and that significant stenosis should be corrected when it is accompanied by graft dysfunction. The rationale for surveillance depends on the dysfunction hypothesis, which states that stenosis causes graft dysfunction [such as a decrease in graft blood flow (Qa)], and this dysfunction reliably precedes and accurately predicts thrombosis. The usefulness of Qa surveillance depends on accurate prediction of thrombosis so that stenosis can be corrected prior to thrombosis. An analysis of the dysfunction hypothesis indicates that some or all of its underlying assumptions are invalid. Most importantly, the presence of wide hemodynamic variation during Qa measurements makes Qa a relatively inaccurate predictor of thrombosis. A number of studies have evaluated the value of surveillance with intervention in reducing thrombosis rates and prolonging graft life. Review of these studies show that few have been prospective and randomized, and many have included historical control groups. It is debatable whether these studies have established that Qa surveillance with intervention should be applied to all grafts. Data from several studies suggest that severity of stenosis may be at least as accurate as Qa in predicting thrombosis. Consequently, inclusion of stenosis measurements (e.g., by duplex ultrasound) may improve the results of surveillance. These unresolved issues indicate it is premature to recommend routine Qa surveillance with intervention of all hemodialysis patients with grafts. PMID- 11422924 TI - Access flow measurement as a predictor of hemodialysis graft thrombosis: making clinical decisions. AB - Since the introduction of dilution methods for measurement of vascular access blood flow during hemodialysis, more than 170 publications addressing the accuracy, prognostic value, and economic impact of the technology have been presented. Recently researchers (Paulson et al.) have raised concerns about the accuracy of access flow measurements in predicting thrombosis. Our first objective was to address the inadequacies of the analysis by these authors. The second objective was to apply a statistically accepted three-step approach for clinical decision making to assess the utility of access flow surveillance (similar to the K/DOQI guidelines) in the prediction of thrombosis. These steps included 1) estimation of treatment thresholds based on harm-benefit analysis of fistulography-angioplasty versus thrombosis, 2) estimation of prior probability of thrombosis based on patient demographic and clinical characteristics, and 3) application of Bayes' theorem to evaluate whether flow test results provided information that could move patients across the treatment threshold, thus discriminating between patients who should be referred for fistulography angioplasty and those who should not. These data and an analysis of recent publications show that the implementation of an access flow surveillance program decreases thrombosis rates in hemodialysis units and can significantly reduce the costs associated with hemodialysis access maintenance. We conclude that access flow monitoring (K/DOQI flow thresholds) is useful in the clinical decision making process for thrombosis prediction across a wide range of demographic categories. PMID- 11422925 TI - Herbs and supplements in dialysis patients: panacea or poison? AB - The safety of herbal remedies and supplement use is of particular concern in patients with renal disease, and reliable information is not always easy to find. Predialysis patients may be drawn to complementary and alternative medicine (CAM) because they believe it can help prevent the progression of their renal disease. The purpose of this series of articles on alternative medicine for nephrologists is to address concerns and issues specific to CAM use in dialysis patients and to provide a guide to reliable sources of information. This introductory article emphasizes safety issues with a focus primarily on herbal medicine. Lack of regulation means that patients may not actually be taking what they think they are. Independent laboratory analyses have shown a lack of stated label ingredients and many instances of supplements and traditional remedies being contaminated with pesticides, poisonous plants, heavy metals, or conventional drugs. While certain supplements are always unsafe (carcinogenic, hepatotoxic, glandular extracts), others are specifically contraindicated in renal disease. Supplement use may be especially hazardous in renal disease because of unpredictable pharmacokinetics, drug interactions, negative effects on kidney function, nephrotoxicity, hemodynamic alterations, unpredictable effects on blood pressure or blood glucose, or potentiation of electrolyte abnormalities. There are no data on potential dialyzability of either active compounds, or their potentially active or toxic metabolites. Many supplements contain metal ions and other minerals. Transplant recipients are also at risk from potential unpredictable effects on immune function. Recommendations and information resources are listed. PMID- 11422926 TI - Dialysis-induced oxidative stress: biological aspects, clinical consequences, and therapy. AB - Oxidative stress, which results from a rupture in the natural balance between pro and antioxidant systems, is considered as a major factor in dialysis-associated morbidity and mortality. Emerging pharmacologic and dialytic antioxidant therapeutic and dialysis strategies should enable us to reduce the harmful consequences of oxidative stress in dialysis patients. Moreover, since there is increasing evidence of oxidative stress long before the initiation of maintenance dialysis, antioxidant therapeutic strategies should probably be developed very early in the course of renal failure. PMID- 11422927 TI - Chronic dialysis and dialysis doctors in the United States: a nephrologist historian's perspective. AB - In an earlier article in Seminars in Dialysis (9:276-281, 1996), the author described the invention of clinical hemodialysis for acute renal failure and its initially equivocal reception by the emerging specialty of nephrology in the United States. A similar story of blunted enthusiasm played out following the invention of the Quinton-Scribner shunt (whose idea "came in the night"), which allowed maintenance treatment for chronic renal failure. Few centers at first could match Belding Scribner's early successes, and some physiology-oriented university nephrologists envisioned how routine dialysis might swamp other activities. Nonetheless, increasingly complex and successful inventions appeared and prevailed: the chronic dialysis unit, the national dialysis chain. A unique federal entitlement program fostered the spread of maintenance dialysis, but so did the emergence of disposable off-the-shelf supplies and many new nephrologists trained in academia but seeking positions in practice. Indeed, the spread of end stage renal disease (ESRD) care transformed American nephrology. The essay concludes by considering what nephrologists of the ESRD era share with their patients. PMID- 11422928 TI - L-carnitine in dialysis patients. AB - Hemodialysis (HD) patients often have low serum concentrations of free L carnitine and decreased skeletal muscle stores. As L-carnitine is an essential cofactor in fatty acid and energy metabolism, it is possible that abnormal carnitine metabolism in dialysis patients may be associated with clinical problems such as skeletal myopathies, intradialytic symptoms, reduced cardiac function, and anemia. Studies have shown that L-carnitine supplementation in HD patients improves several complications seen in dialysis patients, including cardiac complications (arrhythmias, reduced output, low cardiothoracic ratio), limitation of exercise capacity, increased intradialytic hypotension, and muscle symptoms. The most promising results have been noted in the treatment of erythropoietin-resistant anemia. Routine administration of L-carnitine to all dialysis patients is not recommended at this time; however, a therapeutic trial of L-carnitine can be useful in symptomatic patients with certain clinical features unresponsive to the usual measures. These include intradialytic muscle cramps and hypotension, asthenia, cardiomyopathy, lowered ejection fraction, muscle weakness or myopathy, reduced oxygen consumption, and anemia requiring large doses of EPO. PMID- 11422929 TI - Recent advances in immunosuppressive therapy for renal transplantation. AB - Recent advances in immunosuppression have focused on more effective, safer, and targeted therapies that have resulted in improved short- and intermediate-term renal allograft survival. During the past decade there has been a marked decrease in acute rejection rates following renal transplantation because of the use of newer immunosuppressive agents. Recent data indicate that the average yearly reduction in the relative hazard of graft failure beyond 1 year was 4.2% for all recipients (0.4% for those recipients who had an acute rejection episode and 6.3% for those who did not have an acute rejection). Despite these improvements the currently available immunosuppressive agents are associated with significant cardiovascular risk factors, an increased risk of infection, and the development of malignancies in the long term. Predictive parameters of donor-specific hyporesponsiveness are needed so as to allow identification of patients in whom immunosuppressive therapy can be safely reduced. Immunosuppressive agents that have recently been approved for use in the United States and those that are in clinical and preclinical studies are discussed. PMID- 11422930 TI - A case of uremia-associated pleural effusion in a peritoneal dialysis patient. AB - The development of pleural effusions in ESRD patients is a relatively common occurrence. Uremia, currently a rare cause of effusion, can lead to the development of hemorrhagic pleural fluid collections in patients who are underdialyzed. Uremic pleural effusions should be considered when common etiologies of effusions such as volume overload, congestive heart failure (CHF), infection, and malignancy have been excluded. Uremia-associated serosal injury allows transudation of fluid into the pleural space, and defective platelet function contributes to the hemorrhagic nature of the effusion. Although uremic effusions may occur less frequently in peritoneal dialysis patients, we present a patient who developed this complication due to a peritoneal dialysis regimen that was inadequate by current Dialysis Outcomes Quality Initiative guidelines. The pleural effusion resolved with an intensified dialysis regimen in the absence of a change in the underlying volume status. PMID- 11422931 TI - Safety of intravenous iron during infections. PMID- 11422932 TI - Bone mineral density measurements in dialysis patients. PMID- 11422933 TI - New vitamin D analogs. PMID- 11422934 TI - Hepatitis B surface antigen-positive but DNA-negative patients. PMID- 11422936 TI - Effects of different doses in continuous veno-venous haemofiltration on outcomes of acute renal failure: a prospective randomised trial, by Ronco C, Bellomo R, Homel P, Brendolan A, Dan M, Piccinni P, and La Greca G. Lancet 355:26-30, 2000. PMID- 11422937 TI - Sorting and transport of alpha herpesviruses in axons. AB - The alpha herpesviruses, a subfamily of the herpesviruses, are neurotropic pathogens found associated with most mammalian species. The prototypic member of this subfamily is herpes simplex virus type 1, the causative agent of recurrent cold sores in humans. The mild nature of this disease is a testament to the complex and highly regulated life cycle of the alpha herpesviruses. The cellular mechanisms used by these viruses to disseminate infection in the nervous system are beginning to be understood. Here, we overview the life cycle of alpha herpesviruses with an emphasis on assembly and transport of viral particles in neurons. PMID- 11422938 TI - Neurotransmitter receptor trafficking and the regulation of synaptic strength. AB - Modulation of the strength of synapses is thought to be one of the mechanisms that underlies learning and memory and is also likely to be important in processes of neuropathology and drug tolerance. This review focuses on the emerging role of postsynaptic neurotransmitter receptor trafficking as an essential mechanism underlying the dynamic regulation of synaptic strength. PMID- 11422939 TI - Nuclear localization of the Saccharomyces cerevisiae HMG protein NHP6A occurs by a Ran-independent nonclassical pathway. AB - The Saccharomyces cerevisiae non-histone protein 6-A (NHP6A) is a member of the high-mobility group 1/2 protein family that bind and bend DNA of mixed sequence. NHP6A has only one high-mobility group 1/2 DNA binding domain and also requires a 16-amino-acid basic tail at its N-terminus for DNA binding. We show in this report that nuclear accumulation of NHP6A is strictly correlated with its DNA binding properties since only nonhistone protein 6 A-green fluorescent protein chimeras that were competent for DNA binding were localized to the nucleus. Despite the requirement for basic residues within the N-terminal segment for DNA binding and nuclear accumulation, this region does not appear to contain a nuclear localization signal. Moreover, NHP6A does not bind to the yeast nuclear localization signal receptor SRP1 and nuclear targeting of NHP6A does not require the function of the 14 different importins. Unlike histone H2B1 which contains a classical nuclear localization signal, entry of NHP6A into the nucleus was found to be independent of Ran as judged by coexpression of Ran GTPase mutants and was shown to occur at 0 degrees C after a 15-min induction. These unusual properties lead us to suggest that NHP6A entry into the nucleus proceeds by a nonclassical Ran-independent pathway. PMID- 11422940 TI - The mammalian guanine nucleotide exchange factor mSec12 is essential for activation of the Sar1 GTPase directing endoplasmic reticulum export. AB - The Sar1 GTPase is an essential component of COPII vesicle coats involved in export of cargo from the endoplasmic reticulum of mammalian cells. To begin to elucidate its mechanism of action, we now report the identity of the mammalian homolog to the yeast Sec12 guanine nucleotide exchange factor (18% identity) that promotes Sar1 activation. Mammalian Sec12 (mSec12) is a type II transmembrane protein with a large cytosolic domain, a fragment of which has previously been reported as the transcription factor prolactin regulatory element binding protein (PREB). mSec12 promotes efficient guanine nucleotide exchange on Sar1, but not Arf1 or Rab GTPases. mSec12 is localized to the endoplasmic reticulum and an antibody to the cytosolic domain of mSec12 potently inhibits Sar1 recruitment and the formation of COPII vesicles in vitro. The dominant negative GDP-restricted mutant Sar1[T39N] is shown to be a potent inhibitor of mSec12 activity, consistent with its role in preventing COPII vesicle formation in vitro and during transient expression in vivo. We propose that mSec12 is an evolutionarily distant guanine nucleotide exchange factor directing Sar1 GTPase activation in mammalian cells. Its divergence from yeast Sec12p may reflect the specialized needs of the mammalian endoplasmic reticulum involving the formation of Sar1 dependent transitional elements (Aridor M, et al. J Cell Biol 2001;152:213-229) and selection of cargo into prebudding complexes. PMID- 11422941 TI - The class C Vps complex functions at multiple stages of the vacuolar transport pathway. AB - The Class C Vps complex, consisting of Vps11, Vps16, Vps18, and Vps33, is required for SNARE-mediated membrane fusion at the lysosome-like yeast vacuole. However, Class C vps mutants display more severe and pleiotropic phenotypes than mutants specifically defective in endosome-to-vacuole transport, suggesting that there are additional functions for the Class C Vps complex. A SNARE double mutant which is defective for both Golgi-to-endosome and endosome-to-vacuole trafficking replicates many of the phenotypes observed in Class C vps mutants. We show that genetic interactions exist between Class C vps alleles and alleles of the Class D vps group, which are defective in the docking and fusion of Golgi-derived vesicles at the endosome. Moreover, the Class D protein Vac1 was found to physically bind to the Class C Vps complex through a direct association with Vps11. Finally, using a random mutagenic screen, a temperature-conditional allele which shares many of the phenotypes of mutants which are selectively defective in Golgi-to-endosome trafficking was isolated (vps11-3ts). Collectively, these results indicate that the Class C Vps complex plays essential roles in the processes of membrane docking and fusion at both the Golgi-to-endosome and endosome-to-vacuole stages of transport. PMID- 11422942 TI - Cytoplasmic signals mediate apical early endosomal targeting of endotubin in MDCK cells. AB - Endotubin is an integral membrane protein that targets into apical endosomes in polarized epithelial cells. Although the role of cytoplasmic targeting signals as mediators of basolateral targeting and endocytosis is well established, it has been suggested that apical targeting requires either N-glycosylation of the ectoplasmic domains or partitioning of macromolecules into glycolipid-rich rafts. However, we have previously shown that the cytoplasmic portion of endotubin possesses signals that are necessary for its proper sorting into the apical early endosomes. To further define the targeting signals involved in this apically directed event, as well as to determine if the cytoplasmic domain was sufficient to mediate apical endosomal targeting, we generated a panel of endotubin and Tac antigen chimeras and expressed them in Madin-Darby canine kidney cells. We show that both the apically targeting wild-type endotubin and a basolaterally targeted cytoplasmic domain mutant do not associate with rafts and are TX-100 soluble. The cytoplasmic tail of endotubin is sufficient for apical endosomal targeting, as chimeras with the endotubin cytoplasmic domain and Tac transmembrane and extracellular domains are efficiently targeted to the apical endosomal compartment. Furthermore, we show that overexpression of these chimeras results in their missorting to the basolateral membrane, indicating that the apical sorting process is a saturable event. These results show that cells contain machinery in both the biosynthetic and endosomal compartments that recognize cytoplasmic apical sorting signals. PMID- 11422943 TI - Clathrin-dependent and clathrin-independent endocytosis are differentially sensitive to insertion of poly (ethylene glycol)-derivatized cholesterol in the plasma membrane. AB - We examined the effect of a cholesterol derivative, poly (ethylene glycol) cholesteryl ether on the structure/function of clathrin-coated pits and caveolae. Addition of the compound to cultured cells induced progressive smoothening of the surface. Markedly, when the incorporated amount exceeded 10% equivalent of the surface area, fluid pinocytosis, but not endocytosis of transferrin, became inhibited in K562 cells. In A431 cells, both clathrin-independent fluid phase uptake and the internalization of fluorescent cholera-toxin B through caveolae were inhibited with concomitant flattening of caveolae. In contrast, clathrin mediated internalization of transferrin was not affected until the incorporated poly (ethylene glycol) cholesteryl ether exceeded 20% equivalent of the plasma membrane surface area, at which point opened clathrin-coated pits accumulated. The cells were ruptured upon further addition of poly (ethylene glycol) cholesteryl ether. We propose that the primary reason for the differential effect of poly (ethylene glycol) cholesteryl ether is that the bulk membrane phase and caveolae are both more elastic than the rigid clathrin-coated pits. We analyzed the results with the current mechanical model (Rauch and Farge, Biophys J 2000;78:3036-3047) and suggest here that the functional clathrin-lattice is much stiffer than typical phospholipid bilayers. PMID- 11422944 TI - Pathogen inactivation of labile blood components. PMID- 11422945 TI - Pathogen inactivation of labile blood products. PMID- 11422946 TI - Perceptions and motivations of Canadian autologous blood donors. AB - Preoperative autologous blood donation in Canada has increased in the last decade due to concerns about allogeneic blood safety. As economic policies necessitate the validation of autologous blood donation, it is important to assess potential changes. This study examines the motivations and perceptions of patients participating in a Canadian autologous blood donation programme. The study was conducted at the Ottawa Hospital, Civic Campus. Questionnaires were developed and then administered to 100 consecutive adult autologous blood donors. The questionnaires assessed patient demographics, motivation for participation and perceptions about blood safety. Autologous donation was presented to 82% of patients as an option by their physicians. However, 59% of all patients felt that they were motivated to donate by their own fears. Patients (87%) also felt that directed blood donation should be offered due to their perception that directed donation is safer. Three-quarters of patients would be willing to pay for autologous donation. Patients are concerned about the safety of allogeneic blood and they are motivated to seek other alternatives despite the fact that allogeneic blood is very safe. These perceptions and motivations must be taken into account as health care policy changes are considered. PMID- 11422947 TI - The impact of the new tick-box questionnaire, and the personal donor interview, on donor deferrals in the East of Scotland. AB - The impact of a new tick-box questionnaire (TBQ) and personal donor interview (PDI) on donor and recipient safety was assessed over an 18-month and a 13-month period, respectively, by prospectively studying individual donors prior to and after the introduction of the new methodology. A 'hit' was defined as an instance where the TBQ or PDI prompted a donor to divulge information which they would not otherwise have divulged, with the new information having an impact on donor eligibility. There was a 'hit' rate of 0.19% for TBQ and 0.65% for PDI. Of these donors, 33% in the TBQ category and 14% of PDIs were reinstated, 24% and 32%, respectively, were deferred because of a malaria/chagas risk, and 16% of the 'hits' related to donor safety issues. When assessing recipient safety, particularly risk of a window period viral transmission, PDI is very significantly superior at identifying such donors (14 times better). Such information establishes the important safety aspects of these interventions and requires that further work be done to see whether PDIs, in particular, may be better targeted to specific groups of donors. PMID- 11422948 TI - The changes of cardiac ejection fraction up to 72 h after transfusion in patients with chronic anaemia. AB - The objective was to assess the serial change of the cardiac function after blood transfusion in patients with chronic anaemia. According to a previous study, transfusion would adjust the ejection fraction to the advantage of cardiac function in chronic anaemia patients, and thus the serial change of the cardiac function in a certain period should be evaluated after transfusion in patients with chronic anaemia. In a prospective study, ejection fraction (EF) was determined before and < or = 6 h, 24 h and 72 h after transfusion in 32 patients with chronic anaemia, and the results were compared and analysed. In the left ventricle (LV), the four EFs noted before and < or = 6 h, 24 h and 72 h after transfusion did not significantly differ between one another. In the right ventricle (RV), the EF 72 h after transfusion significantly increased in comparison with the pretransfusion value (P = 0.043). Transfusion of small amounts of RBC did not significantly change the LVEF up to 72 h in patients with transfusion-dependent chronic anaemia; whether it might change the RVEF 72 h after transfusion deserves further investigation. PMID- 11422949 TI - Comparison of two platelet additive solutions. AB - The use of an additive solution for substitution of plasma for storage of leukodepleted platelet concentrates can have many advantages. In this study, a comparison was made between two platelet additive solutions: one containing citrate and acetate (PAS-II), the other also supplemented with additional salts such as magnesium, and with gluconate (Composol-PS). Donor-dependent differences were avoided by applying a paired experimental design (n = 10). The platelet concentrates were prepared by pooling five buffy coats and the additive solution, and prestorage filtration was utilized to remove leucocytes to well below 1 x 106. Storage of platelet concentrates up to 9 days after blood collection revealed that platelet concentrates in Composol-PS maintained an almost constant pH of on average 6.93 from day 2 through day 7, and at 6.90 at day 9. This was in contrast to PAS-II, which showed a gradually decreasing pH from on average 6.97 at day 1 to 6.86 at day 9. In all units stored in both solutions the swirling effect was present during 9 days of storage. In conclusion, both additive solutions allow storage of platelets, derived from pooled buffy coats, for up to 9 days after collection of the whole blood, with maintenance of good quality in vitro. Composol-PS has a slightly better buffering capacity, reflected as a more constant pH throughout the storage period. PMID- 11422950 TI - Improved platelet compatiblity of water vapour glow discharge treated non-woven poly(ethylene terephthalate) leukocyte-reduction filters for different types of platelet concentrates. AB - Non-woven poly[ethylene terephthalate] (NW-PET) filter fabric, usually used for leucocyte removal of red cells, was modified by water vapour glow discharge (WVGD) treatment to improve platelet compatibility. Modified filter material was evaluated with different kinds of platelet concentrates (PCs). In addition, modified filter materials were gamma-sterilized and tested after different time intervals at different storage conditions. Modification of the filter material resulted in an improved platelet recovery after filtration of PC from 57 to about 80%. No significant difference in platelet recovery was observed when filtering either freshly prepared (79 +/- 3.5%, mean +/- SD), overnight-stored single BC-PC (78 +/- 3.3%), overnight-stored single PRP-PC (75 +/- 8.8%) or overnight-stored pooled BC-PC (79 +/- 8.9%). However, freshly prepared pooled BC-PC gave a significantly higher platelet recovery (84 +/- 3.5%). Leukocyte depletion did not differ significantly between the different types of PC. gamma-Sterilization and subsequent storage of the modified filter material for 5, 14 and 26 weeks at 20 degrees C or 37 degrees C had no significant influence on the filtration results of overnight-stored pooled BC-PC. The results of the present study show that WVGD treated NW-PET is platelet compatible and can be used for leucocyte removal from preferably BC-PC. It can be gamma-sterilized and stored for at least 6 months prior to filtration without affecting the platelet recovery and leucocyte removal. PMID- 11422951 TI - The frequency of anti-C + anti-G in the absence of anti-D in alloimmunized pregnancies. AB - Anti-D+C are often initially identified in sera from alloimmunized women. Anti-G may be present in these samples, mimicking anti-D+C, and therefore the differentiation of anti-D, -C and -G may be important. Sera from 27 alloimmunized women, initially identified as containing anti-D + anti-C, were analysed by adsorption/elution studies in the presence of polyethylene glycol (PEG) using R(0)r (D+C-G+) and r'r(D-C+G+) red blood cells (RBC). Additionally, 15/27 samples were tested by adsorption in the presence of PEG and subsequently warm elution, using rGr (D-C-G+) RBC. Anti-G + anti-C, without anti-D, were identified in 4/27 samples (14.8%) and none of the newborn children needed postpartum treatment. The combination of D+G, D+C and D+C+G antibodies occurred in 25.9%, 11.1% and 48.1% of the women, respectively. Overall, anti-G was detected in 24/27 samples (88.9%). Pregnant women shown to have anti-G+C but not anti-D should receive Rh immune globulin. Additionally, the finding of apparent anti-D+C during pregnancy in D-negative spouses may lead to paternity testing and therefore a correct antibody identification is necessary. PMID- 11422952 TI - Workshop report on the genotyping of blood cell alloantigens. AB - The immunization against alloantigens present on platelets, granulocytes and red blood cells (RBCs) is responsible for various clinical syndromes. Since the molecular basis of these antigens has become clear during the last decade, genotyping is nowadays used in several laboratories. However, many DNA-based techniques still have to be evaluated. We therefore organized a workshop on the genotyping of the most relevant alloantigens on platelets and granulocytes as well as on selected RBC alleles. DNA was isolated from peripheral blood lymphocytes or from B-lymphoblastoid cell lines (B-LCL). We distributed samples for the identification of platelet (n = 7), granulocyte (n = 6) and RBC (n = 4) polymorphisms, respectively. There were 33 institutions in Germany, Austria and Switzerland, which participated in at least one part of the workshop. Twenty-four laboratories reported results on HPA-1, and 23 laboratories on HPA-2, -3, and -5 typing. In addition, five laboratories typed for HPA-4 and -6. The HNA-1a/b (NA1/NA2) alleles were identified by eight laboratories, one of which also typed for HNA-1c (SH). The most frequent genes of the ABO (A1, B, O) and Rh (D, C, c, E, e) systems were typed by 12 participating laboratories, and an additional four laboratories restricted their RBC typing to the RHD gene. The typing technique mainly used for all three cell lineages was the polymerase chain reaction with sequence-specific primers. Other techniques were restriction fragment length analysis, oligonucleotide ligation assay, enzyme-linked mini-sequence assay or direct sequence analysis. The following typing errors were observed: HPA: 15/1442 (1.0%), HNA: 4/108 (3.7%), ABO: 5/96 (5.2%) RH 1/320 (0.3%). Our workshop demonstrated the existence of a number of reliable techniques for the genotyping of blood cell alloantigens and a high standard in the participating laboratories. In addition, we could show the usefulness of B-LCL as a source of reference DNA. However, the 5.2% rate of mistyping in the ABO system demonstrated that further efforts are needed to improve the precision of the genotyping techniques. Future workshops will have to challenge methods and participants with rare variants of RBC genes to guarantee reliable genotyping, e.g. in prenatal diagnosis of fetomaternal incompatibility. PMID- 11422953 TI - The BCSH guideline on addressograph stickers: an update. PMID- 11422954 TI - Affordable antiretroviral drugs for developing countries: dreams of the magic bullet. PMID- 11422955 TI - Diagnostic accuracy and case management of clinical malaria in the primary health services of a rural area in south-eastern Tanzania. AB - Malaria control continues to rely on the diagnosis and prompt treatment of both suspected and confirmed cases through the health care structures. In south eastern Tanzania malaria is one of the leading causes of morbidity and mortality. The absence of microscopic examination in most of the health facilities implies that health workers must rely on clinical suspicion to identify the need of treatment for malaria. Of 1558 randomly selected paediatric consultations at peripheral health facilities throughout Kilombero District, 41.1% were diagnosed by the attending health worker as clinical malaria cases and 42.5% prescribed an antimalarial. According to our malaria case definition of fever or history of fever with asexual falciparum parasitaemia of any density, 25.5% of all children attending the health services had malaria. This yielded a sensitivity of 70.4% (IC95% = 65.9-74.8%) and a specificity of 68.9% (IC95% = 66.2-71.5%). Accordingly, 30.4% of confirmed cases left with no antimalarial treatment. Among malaria-diagnosed patients, 10% were underdosed and 10.5% were overdosed. In this area, as in many African rural areas, the low diagnostic accuracy may imply that the burden of malaria cases may be overestimated. Greater emphasis on the functioning and quality of basic health services in rural endemic areas is required if improved case management of malaria is to help roll back this scourge. PMID- 11422956 TI - Therapeutic efficacy of sulphadoxine/pyrimethamine and susceptibility in vitro of P. falciparum isolates to sulphadoxine-pyremethamine and other antimalarial drugs in Malawian children. AB - Since 1993 sulphadoxine/pyrimethamine (SP) has been used as the first-line drug for uncomplicated Plasmodium falciparum malaria in Malawi. To investigate the current efficacy of SP and other antimalarial drug resistance, we studied in vivo and in vitro responses to SP, chloroquine (CQ), mefloquine (MF), quinine (QN), and halofantrine (HF) in Salima, central Malawi. In a follow-up of 14 days, nine (13.8%) of 65 children under five showed RII/RIII parasitological resistance, and in in vitro microtests 18 (62.1%) of 29 isolates showed < 90% inhibition of schizont maturation at pyrimethamine 75 nmol/l blood medium mixture, indicating resistance. The discrepancy between in vivo and in vitro results might be partially explained by acquired immunity in this holoendemic area. In vitro one (3.4%) of 29 isolates failed schizont inhibition at 1.6 micromol/l blood of CQ, indicating resistance. Compared with an in vitro study conducted in 1988 in another region of Malawi using the same cut-off point, the proportion of resistant isolates had decreased significantly (P < 0.01). Although 31% of isolates were borderline, showing schizont maturation at 0.8 micromol/l blood but no schizonts at 1.6 micromol/l in our study, the results suggest possible recovery of CQ sensitivity after long-term absence of drug pressure. Resistance remains a major problem in malaria control. Monitoring resistance patterns in vitro provides early warning signs of impending loss of therapeutic efficacy of the standard treatment, and may detect changing patterns in alternative drug resistance. PMID- 11422957 TI - Enumeration of non-communicable disease in rural South Africa by electronic data linkage and capture-recapture techniques. AB - Non-communicable diseases (NCDs) are becoming increasingly common and important in developing countries, yet their enumeration is problematic. We have attempted to enumerate NCD patients in a rural district of KwazuluNatal, South Africa, using the techniques of electronic data linkage and capture-recapture (CR). We examined four major NCDs (hypertension, diabetes, asthma and epilepsy). Basic patient details were recorded onto EpiInfo software over a 6-week period, from the main hospital clinic at Hlabisa, as well as the 10 outlying peripheral health clinics. Using electronic data linkage of lists from the main hospital, the peripheral clinics, and repeat prescription cards, a district NCD register was produced of 2455 patients. The mean age was 51 +/- 16 years (1 SD) and 76% were female. Of the total NCD patients, 62% had hypertension (age 57 +/- 12 years, 82% female), 16% epilepsy (age 35 +/- 17 years, 49% female), 13% asthma (age 45 +/- 19 years, 60% female) and 12% diabetes (age 54 +/- 13 years, 61% female). Estimated population crude prevalence rates for known NCD cases on the register were 7.4% for hypertension, epilepsy 0.2%, asthma 0.2% and diabetes 0.2%. We also attempted a CR analysis to assess completeness of data, by comparing overlap between patients attending peripheral clinics, and the central Hlabisa Hospital clinic. Matching by name, age, and diagnosis proved feasible, but there was little overlap, and CR calculations were invalid because of the relative independence of sources. We conclude that NCDs are common in rural Africa, and that a simple NCD district register is a potentially feasible and inexpensive option. Capture-recapture analysis is feasible, but requires suitable lists with acceptable overlap of patients. PMID- 11422958 TI - Community perceptions of a mass administration of an antimalarial drug combination in The Gambia. AB - To test the hypothesis that widespread treatment with artemisinin derivatives can reduce malaria transmission, a mass drug administration (MDA) campaign was undertaken in an area of The Gambia in 1999. Coverage of 85% of the target population was achieved, but the intervention did not reduce overall malaria transmission. We studied the perceptions, knowledge and attitudes of the community to the MDA campaign. A validated questionnaire was administered to randomly selected MDA participants (n = 90) and MDA refusers (n = 71). Individuals who believed in the importance of the MDA (adjusted OR 58.3%; 95% CI 17.4-195.8) and those who were aware that a high level of participation was needed for the MDA to be successful (adjusted OR 28.1; 95% CI 10.3-75.9) were more likely to participate. Understanding that the purpose of the MDA was to reduce malaria (adjusted OR 13.9; 95% CI 5.5-35.1) and knowledge of the fact that malaria is transmitted by mosquitoes and of the clinical signs of malaria (adjusted OR 3.4; 95% CI 3.1-9.0) were associated with participation. Individuals who discussed the MDA with other villagers (adjusted OR 5.5; 95% CI 2.2-13.5) and those who attended the sensitization meeting (adjusted OR 2.6; 95% CI 1.1-6.0) were also more likely to participate. Women were significantly more likely to participate in the MDA than men (adjusted OR 3.1; 95% CI 1.5-6.2). Individuals who refused to participate were unlikely to plan participation in future MDAs. One of the most difficult challenges in the implementation of a malaria control strategy such as an MDA is to convince villagers to participate and to make them aware that a high level of participation by the community is needed for success. We found that our sensitization meetings could be improved by giving more information on how the MDA works and finding means to generate small group discussions after the meeting. PMID- 11422959 TI - Intestinal helminth infections, anaemia and labour productivity of female tea pluckers in Bangladesh. AB - We conducted a randomized clinical intervention trial over 24 weeks on a tea estate in north-east Bangladesh to investigate the effect of iron supplementation and anthelmintic treatment on the labour productivity of adult female tea pluckers. A total of 553 full-time tea pluckers, not pregnant and not breastfeeding, were randomly assigned to one of the four intervention groups: group 1 received iron supplementation on a weekly basis, group 2 received anthelmintic treatment at the beginning and half-way through the trial (week 12), group 3 received both iron supplementation as group 1 and anthelmintic treatment as group 2, and group 4 was a control group and received placebos. No significant difference in labour productivity was found between the four intervention groups over the trial period. However, there was a negative association for all three worms (Ascaris lumbricoides, Trichuris trichiura and hookworms) between the intensity of helminth infections (eggs/g faeces) and all measures of labour productivity. Lower haemoglobin values and anaemia (< 120 g/l Hb) were both associated with lower labour productivity and more days sick and absent. Taller women with greater arm circumference were able to pluck more green leaves, earn higher wages and were absent less often. PMID- 11422960 TI - PCR-based ELISA technique for malaria diagnosis of specimens from Thailand. AB - We performed a field evaluation of polymerase chain reaction (PCR)-based enzyme linked immuno-sorbent assays (ELISA) for the diagnosis of malaria. A commercially available PCR-ELISA microplate hybridization (MPH) assay was used. Blood specimens were collected from 300 volunteers seeking care at malaria clinics in Thailand. Examination of 200 high power fields by Giemsa-stained thick and thin smear (GTTS) revealed 51 P. falciparum (Pf), 45 P. vivax (Pv), seven mixed Pf-Pv infections. These plus a random sample of 48 GTTS-negative specimens were selected for this study. All 151 specimens were processed for parasite DNA extraction and assayed by PCR-MPH. The target DNA sequence of the 18S small subunit ribosomal RNA (SSUrRNA) gene was amplified by PCR and hybridized with species-specific probes for Pf, Pv, P. malariae (Pm) and P. ovale (Po) immobilized in the wells of the microtiter plate and detected by colorimetric assay. Colour development was assessed at an optical density (OD) of 405 nm. An absorbance reading of > or = 0.1 was used as a positive cut-off. In comparison with GTTS results, PCR-MPH sensitivity was 91.4% (53/58, 95% CI 84.2-98.6) for Pf, 94.2% (49/52, 87.9-100) for Pv and specificity was 95.8% (46/48, 95% CI 90.2 100). There was statistically significant positive correlation between parasite densities < or = 7000/microl blood and absorbance reading, suggestive of PCR-MPH being semiquantitative. PCR-MPH also detected additional Pf and Pv cases as well as Pm and Po. PMID- 11422961 TI - Molecular tools for the diagnosis of cystic and alveolar echinococcosis. AB - In the past years, the diagnostic tools applied to identify alveolar (AE) and cystic echinococcosis (CE) in human patients have not only increased in number but also substantially improved in quality. The identification and characterization of species-specific parasite proteins/antigens allowed to generate subsequently recombinant or synthetic polypeptide antigens, as well as corresponding monoclonal antibodies. Some of these new tools have already demonstrated operating characteristics superior to conventional tests used for the immunodiagnosis of CE and AE, and thus may be suggested for routine laboratory application. Powerful molecular techniques, such as the polymerase chain reaction (PCR), have been developed and adapted to advance laboratory diagnosis of AE and CE. Detecting minute amounts of parasite DNA and mRNA, not only to identify but also to characterize the biological status of parasite material, thus becomes a complementary method to synergize immunodiagnostic techniques. This review focuses on recent developments of molecular tools, discussing their potential use as a primary or a supporting diagnostic element. We also outline some future developments to be undertaken in the field of molecular diagnosis, linked to clinical and laboratory problems. PMID- 11422962 TI - Predictors of unfavourable prognosis in chronic Chagas' disease. AB - The aim of this study was to detect clinical predictors of left ventricular dysfunction, left ventricular dilatation and apical aneurysm on echocardiography, all known as independent predictors of lethal outcome for patients with chronic Chagas' disease. Seventy-four patients with a positive complement-fixation test for Chagas' disease participated; 44 (59%) had left ventricular dysfunction, 41 (55%) left ventricular dilatation and 15 (20%) apical aneurysm. A stepwise logistic regression analysis showed that systolic blood pressure (P < 0.001) and male sex (P < 0.001) were independent predictors of left ventricular dilatation on echocardiography. A receiver-operating characteristic curve provided a systolic blood pressure of 120 mmHg with a sensitivity of 70% and a specificity of 63% to predict left ventricular dilatation. The combination of male sex and systolic blood pressure of 120 mmHg had a sensitivity of 56% and a specificity of 91% to predict left ventricular dilatation. In a separate stepwise logistic regression analysis, left ventricular systolic dysfunction was independently predicted by systolic blood pressure (P = 0.006) and New York Heart Association functional class (P = 0.01). Receiver-operating curves provided a blood pressure of 120 mmHg with a sensitivity of 72% and a specificity of 59% to predict left ventricular dysfunction, whereas a New York Heart Association functional score of 2 predicted left ventricular systolic dysfunction with a sensitivity of 78% and a specificity of 50%. The combination of New York Heart Association functional class and a systolic blood pressure of 120 mmHg predicted left ventricular dysfunction with a sensitivity of 59% and a specificity of 77%. The apical aneurysm was independently predicted by myocardial necrosis on the resting ECG, but only with a sensitivity of 20%. Hence, echocardiographic markers of cardiac mortality and sudden cardiac death in Chagas' disease can be independently predicted by clinical examination. This may be useful for screening high-risk chagasic patients. PMID- 11422963 TI - Poor food hygiene and housing as risk factors for typhoid fever in Semarang, Indonesia. AB - To identify risk factors for typhoid fever in Semarang city and its surroundings, 75 culture-proven typhoid fever patients discharged 2 weeks earlier from hospital and 75 controls were studied. Control subjects were neighbours of cases with no history of typhoid fever, not family members, randomly selected and matched for gender and age. Both cases and controls were interviewed at home by the same trained interviewer using a standardized questionnaire. A structured observation of their living environment inside and outside the house was performed during the visit and home drinking water samples were tested bacteriologically. Univariate analysis showed the following risk factors for typhoid fever: never or rarely washing hands before eating (OR = 3.28; 95% CI = 1.41-7.65); eating outdoors at least once a week (OR = 3.00; 95% CI = 1.09-8.25); eating outdoors at a street food stall or mobile food vendor (OR = 3.86; 95% CI = 1.30-11.48); consuming ice cubes in beverage in the 2-week period before getting ill (OR = 3.00, 95% CI = 1.09-8.25) and buying ice cubes from a street vendor (OR = 5.82; 95% CI = 1.69 20.12). Water quality and living environment of cases were worse than that of controls, e.g. cases less often used clean water for taking a bath (OR = 6.50; 95% CI = 1.47-28.80), for brushing teeth (OR = 4.33; 95% CI = 1.25-15.20) and for drinking (OR = 3.67; 95% CI = 1.02-13.14). Cases tended to live in houses without water supply from the municipal network (OR=11.00; 95% CI = 1.42-85.2), with open sewers (OR = 2.80; 95% CI = 1.0-7.77) and without tiles in the kitchen (OR = 2.67; 95% CI = 1.04-6.81). Multivariate analysis showed that living in a house without water supply from the municipal network (OR = 29.18; 95% CI = 2.12-400.8) and with open sewers (OR = 7.19; 95% CI = 1.33-38.82) was associated with typhoid fever. Never or rarely washing hands before eating (OR = 3.97; 95% CI = 1.22 12.93) and being unemployed or having a part-time job (OR = 31.3; 95% CI = 3.08 317.4) also were risk factors. In this population typhoid fever was associated with poor housing and inadequate food and personal hygiene. PMID- 11422964 TI - Toxoplasmosis: will the time ever come? PMID- 11422965 TI - Systematic reviews of trials of umbilical artery Doppler: time for more primary research. PMID- 11422966 TI - A critical appraisal of the use of umbilical artery Doppler ultrasound in high risk pregnancies: use of meta-analyses in evidence-based obstetrics. AB - OBJECTIVES: To reanalyze randomized controlled trials on the use of umbilical artery Doppler velocimetry in high-risk pregnancies and determine which high-risk pregnancies benefit from the use of Doppler velocimetry. METHODS: Searching Medline, the Cochrane Library and Embase we found 13 randomized controlled trials on the use of Doppler velocimetry in high-risk pregnancies. Of these, six included pregnancies with strictly defined suspected intrauterine growth restriction and/or hypertensive disease of pregnancy ('well-defined studies'); the rest included a great variety of high-risk pregnancies ('general risk studies'). The studies were analyzed with particular regard to the heterogeneity and to outcome. Audits of the perinatal deaths reported in the randomized controlled trials were performed by a panel of 32 international experts. RESULTS: The 'well-defined studies' had a more uniform study design as compared to the 'general risk studies' and they showed a significant reduction in antenatal admissions (odds ratio, 0.56; 95% confidence interval, 0.43-0.72), inductions of labor (0.78; 0.63-0.96), elective deliveries (inductions of labor and elective Cesarean sections) (0.73; 0.61-0.88) and Cesarean sections (0.78; 0.65-0.94). By perinatal audit it was found that more perinatal deaths in the 'well-defined studies' were potentially avoidable by use of Doppler velocimetry ( P < 0.0005) and the rate of avoidable perinatal deaths was higher among controls (50%) than cases (20%) in this group. CONCLUSION: The randomized controlled trials on umbilical artery Doppler velocimetry show major differences regarding study design and technical and clinical issues and, therefore, they should not be pooled in a simple meta-analysis. By stratification it was found that only in pregnancies with suspected intrauterine growth restriction and/or hypertensive disease of pregnancy will the use of umbilical artery Doppler velocimetry reduce the number of perinatal deaths and unnecessary obstetric interventions. PMID- 11422967 TI - Improved early prediction of pre-eclampsia by combining second-trimester maternal serum inhibin-A and uterine artery Doppler. AB - OBJECTIVE: To evaluate the screening efficacy of second-trimester maternal serum inhibin-A combined with uterine artery Doppler studies performed at 20 weeks' gestation for the prediction of pre-eclampsia. METHODS: Six hundred and eighty nine consecutive unselected women who had inhibin-A levels measured between 15 and 19 weeks' gestation subsequently had color flow pulsed Doppler of both uterine arteries at the time of the anomaly scan (mean 20.5, range 18-22 weeks' gestation). The main outcome measures were pre-eclampsia and pre-eclampsia requiring delivery before 37 weeks' gestation. The presence or absence of notches in the flow velocity waveform was noted, and the resistance index measured. Receiver operator curves were created for inhibin-A and uterine artery Dopplers alone and in combination. Sensitivities for each method were compared at false positive rates of 3% and 7%. RESULTS: Thirty-five women developed pre-eclampsia, of whom 15 required delivery before 37 weeks' gestation. For a false-positive rate of 7%, the sensitivity using bilateral notches/mean resistance index > or = 0.65 was 60% with a positive likelihood ratio of 8.6 (confidence interval 5.7 12.6). For the same false-positive rate, when bilateral notches/mean resistance index > or = 0.55 and unilateral notches/mean resistance index > or = 0.65 were combined with inhibin-A > or = 1.0 multiples of the median, the sensitivity improved to 71% and the positive likelihood ratio to 10.8 (confidence interval 7.4-15.4). For pre-eclampsia requiring delivery before 37 weeks for a false positive rate of 3%, the sensitivity for bilateral notches improved from 27% to 60% and the positive likelihood ratio improved from 9.2 to 20.8 when uterine artery notch data was combined with inhibin-A. The improvement in sensitivity for the combined method, compared to either inhibin-A or uterine artery Dopplers alone, was statistically significant for both pre-eclampsia ( P < 0.05) and preterm pre-eclampsia ( P < 0.02). CONCLUSION: Combination of second-trimester serum inhibin-A and uterine artery Doppler at 20 weeks improves the screening efficacy for the prediction of pre-eclampsia especially when this necessitates early delivery. This combination might form the basis of an effective and practical early screening test for the condition. PMID- 11422968 TI - Three-dimensional (3D) echocardiographic analysis of congenital heart disease in the fetus: comparison with cross-sectional (2D) fetal echocardiography. AB - OBJECTIVE: We attempted to assess the ability of Doppler-gated three-dimensional (3D) fetal echocardiography to reconstruct and display specific cardiac structures in fetuses with cardiac anomalies and to determine whether any advantage is offered by 3D sonographic cardiac examination over conventional fetal echocardiography. DESIGN: After 2D fetal echocardiographic examination, 3D cardiac data were collected prospectively in 22 fetuses with various congenital heart defects. Their ages ranged from 19 to 35 weeks' gestation. Basic echocardiographic key views of the venoatrial, atrioventricular and ventriculoarterial connections were derived from volume data sets and selected for 3D reconstruction and analysis. Comparisons were made with 2D echocardiographic imaging of the fetal hearts and the diagnostic image quality of visualized structural details was evaluated. RESULTS: The underlying cardiac malformation was well or satisfactorily visualized in 20 fetuses using 2D imaging. Gated 3D volume data sets enabled diagnostically acceptable visualization of all affected cardiac structures in 7 of 22 fetuses. High-quality 3D reconstruction of the site and spatial orientation of ventricular septal defects was obtained in 9 of 13 patients. Two-dimensional imaging remained the principal diagnostic modality in all cases with additional structural detail being obtained by 3D imaging in only two fetuses. CONCLUSIONS: Three-dimensional imaging of fetal heart disease is feasible for a wide range of lesions, and may provide additional information of clinical value in a small number of cases when compared with 2D imaging. PMID- 11422969 TI - Abdominal circumference: a single measurement versus growth rate in the prediction of intrapartum Cesarean section for fetal distress. AB - OBJECTIVE: The fetal abdominal circumference is the most sensitive ultrasound biometric measurement for predicting intrauterine growth restriction, which is associated with an increased risk of intrapartum fetal distress. We sought to evaluate and compare whether a third-trimester ultrasound measurement of abdominal circumference made within 1 week prior to delivery better predicts operative delivery for fetal distress when compared with the growth velocity of the abdominal circumference in the third trimester. METHODS: Retrospective analysis was carried out of prospectively collected ultrasound data on 117 patients with singleton gestations who had had at least two ultrasound assessments performed less than 6 weeks apart in the third trimester, with the last ultrasound performed within 1 week prior to delivery. The abdominal circumference value of the last ultrasound prior to delivery was placed into one of three categories: < or = 5% centile, > 5 to < or = 10% centile and > 10% centile for gestational age. The growth velocity of the abdominal circumference per week was placed into one of three categories: < or = 5 mm/week, 6-10 mm/week and > or = 11 mm/week. The chi-squared test was used to compare differences between the incidence of fetal distress between the groups. RESULTS: The incidences of Cesarean section for fetal distress in relation to a single measurement of the abdominal circumference were: < or = 5% centile, 8/23 (35%); > 5 to < or = 10% centile, 3/12 (25%); > 10% centile, 8/81 (10%) ( P < 0.05). The incidences of Cesarean section for fetal distress with the three abdominal circumference growth velocities were: < or = 5 mm/week, 9/55 (16%); 6-10 mm/week, 4/11 (36%); > or = 11 mm/week, 8/51 (16%) ( P = 0.9401). CONCLUSION: A single measure of the fetal abdominal circumference made within 1 week prior to delivery is superior to an assessment of growth rate of the fetal abdomen in the third trimester in discriminating patients who require Cesarean section for fetal distress. PMID- 11422970 TI - The relevance of placental location at 20-23 gestational weeks for prediction of placenta previa at delivery: evaluation of 8650 cases. AB - OBJECTIVE: To determine the correlation between placental position at 20-23 weeks and incidence of birth complications caused by placental position. SUBJECTS AND METHODS: In an ongoing prospective study, placental position was determined by transabdominal sonography as part of anomaly scanning at 20-23 gestational weeks, followed by transvaginal sonography in uncertain or suspicious situations. Examination was performed in 9532 cases; feedback was obtained from 8650 patients (90.7%). RESULTS: Transabdominal sonography was followed by transvaginal scan in 363 of 8650 cases (4.2%). In 8551 cases (98.9%), we found normal placental position, with the placenta not reaching the internal os and a Cesarean section rate of 17.1% (1458/8551). The incidence of 'low placental position', with the placenta reaching the internal os was 0.66% (57/8650), with a Cesarean section rate of 21% (12/57). In 0.49% (42/8650) of cases, the placenta overlapped the internal os at 20-23 weeks; Cesarean section because of placenta previa or bleeding was performed in 28 of 8650 cases (0.32%). Vaginal delivery was possible in 43% of cases (13/30), when the overlap did not exceed 25 mm. If the overlap exceeded 25 mm (12 cases), no vaginal delivery was reported. There was no reported case of placenta previa missed at the 20-23-week scan. CONCLUSION: At 20 23 weeks, a combination of routine transabdominal and indication-based transvaginal location of placental position is a powerful tool in predicting placenta previa at delivery. The advantage of determining placental position at this stage of pregnancy is a low false-positive rate compared to at earlier stages of pregnancy. We conclude that an overlapping placenta at 20-23 weeks has the consequence of a high probability of placenta previa at delivery. An overlap of 25 mm or more at 20-23 weeks seems to be incompatible with later vaginal delivery. PMID- 11422971 TI - Fetal transcerebellar diameter and chromosomal abnormalities. AB - OBJECTIVE: To evaluate the association between chromosomal abnormalities and fetal cerebellar size. DESIGN: A retrospective cross-sectional study. METHODS: Ultrasound measurements of transcerebellar diameter, head and upper-abdominal circumference from 88 fetuses with chromosomal abnormalities were analyzed. Abnormalities included trisomy 21 ( n = 23), trisomy 18 ( n = 17), 'other numerical chromosomal abnormalities' ( n = 9), sex chromosomal abnormalities ( n = 9), mosaicism ( n = 12), balanced translocations ( n = 9) and unbalanced translocations ( n = 9). Multiple regression analysis was performed to compare transcerebellar diameters between the reference group and each of the subsets of chromosomal abnormalities and between trisomies 18 and 21. Also, in the latter two subsets, comparison of the transcerebellar diameter before and after 25 weeks of gestation was carried out. RESULTS: Fetal transcerebellar diameter was reduced in relation to gestational age but was normal when control was made for fetal size in all chromosomal subsets, except for balanced translocations. The transcerebellar diameter in trisomy 18 was significantly smaller than that in trisomy 21. No difference in cerebellar size was found when comparing the gestational age period before and after 25 weeks in each of these two subsets. CONCLUSIONS: A reduction in fetal transcerebellar diameter was demonstrated in all chromosomal abnormalities with imbalance of genetic material. Cerebellar hypoplasia was more severe in trisomy 18 than in trisomy 21. The degree of reduction in fetal transcerebellar diameter in these subsets seems to be independent of the time period during which the transcerebellar diameter measurement was performed. PMID- 11422972 TI - The role of ultrasound in the expectant management of early pregnancy loss. AB - OBJECTIVE: To define the sonographic criteria which best determine the likelihood of successful expectant management of early pregnancy failure (EPF). METHODS: Women with an ultrasound diagnosis of EPF at 7-14 weeks' gestation were offered the option of expectant management or surgical evacuation. RESULTS: Five hundred and forty-five women had a diagnosis of EPF; 298 with incomplete miscarriage and 247 with missed miscarriage or an embryonic pregnancy. A total of 305 women opted for expectant management, with an overall success rate of 86%. The success rate for incomplete miscarriage (96%) was significantly better than that for missed miscarriage (62%). CONCLUSION: This study demonstrates that EPF can be safely managed expectantly. Ultrasound has an invaluable role in predicting the likelihood of successful expectant management enabling patients to make an informed choice about their medical care. PMID- 11422973 TI - The significance of crown-rump length measurement for predicting adverse pregnancy outcome of threatened abortion. AB - OBJECTIVES: To determine if in women with threatened miscarriage the measurement of fetal crown-rump length (CRL) is a useful predictor of spontaneous miscarriage and small for gestational age (SGA) infants. METHODS: Fetal CRL was measured in 310 singleton pregnancies with live fetuses, presenting with threatened miscarriage before 13 weeks of gestation. The relationship between fetal CRL and pregnancy outcome was investigated. RESULTS: In fetuses with CRL < 18 mm there was a significant positive association between the deficit in CRL for gestation and the incidence of subsequent spontaneous miscarriage. In those cases with CRL deficit more than 2 standard deviations (SDs) the incidence of miscarriage was 13.7%, whereas if the CRL was between the mean and -2 SDs the incidence of miscarriage was 8.3%. In fetuses with CRL > or = 18 mm there was a significant positive association between the deficit in CRL for gestation and the incidence of SGA. In those cases with CRL deficit of more than 2 SD the incidence of SGA was 27.3%, whereas if the CRL was between the mean and -2 SDs the incidence of SGA was 18.9%. CONCLUSIONS: The findings of this study suggest that the measurement of fetal CRL may be a useful predictor of spontaneous miscarriage and SGA in pregnancies with threatened miscarriage. PMID- 11422974 TI - Sonographic early fetal gender assignment: a longitudinal study in pregnancies after in vitro fertilization. AB - OBJECTIVES: A longitudinal evaluation by sonography of external genitalia in human embryos/early fetuses with a known time from fertilization is lacking. Our aim was to assign by sonographic evaluation of external genitalia the early fetal gender in a cohort of pregnancies after in vitro fertilization. Sonographic examinations were performed in each case in three subsequent sessions over a period of time early in gestation in order to establish a temporal threshold, expressed in terms of days from fertilization, at which absolute accuracy in gender prediction is achievable. METHODS: Thirty-two fetuses were included in this prospective longitudinal study. Each was examined three times for gender assignment. The first observation was performed between 65 and 69 days from fertilization, the second between 70 and 74 days and the third between 75 and 79 days. Transvaginal and/or transabdominal sonography was used to detect the 'sagittal sign' as a marker of fetal gender. The results of ultrasound examinations were compared with gender at birth or with karyotype obtained from amniotic fluid cells or chorionic villus sampling. RESULTS: Fetal gender assignment was feasible in 29 out of 32 fetuses (90%) at the first examination and in all cases at the second and third examinations. Fetal gender prediction was correct in 76% of cases in which fetal gender was assigned (22/29) at the first examination; accuracy for males was 46% (6/13) and for females 100% (16/16). At the second and third examinations, accuracy for gender prediction achieved 100% for both genders. Concerning the temporal threshold, absolute accuracy in gender prediction was achieved at 69 days from fertilization, corresponding to 11+6 weeks based on the last menstrual period. CONCLUSION: This study provides important information about the earliest stage, expressed in terms of days from fertilization, at which it is possible to make a certain diagnosis of fetal gender by sonography. PMID- 11422975 TI - The feasibility of a 'one stop' ultrasound-based clinic for the diagnosis and management of abnormal uterine bleeding. AB - OBJECTIVES: We have established a 'one stop' clinic for the management of women with abnormal uterine bleeding based on transvaginal sonography and saline contrast sonohysterography. This report reviews our experience with the first 93 patients attending the clinic. DESIGN AND METHODS: Patients were seen with the intention of performing a transvaginal scan, saline contrast sonohysterography, endometrial biopsy, full blood count and thyroid function tests. The findings were prospectively recorded on a computer database and a management plan formulated. RESULTS: A transvaginal scan was performed on 89 (95.7%) women, 70 (75.3%) also undergoing saline contrast sonohysterography. An endometrial biopsy was carried out in 67 (72%) women aged 40 years and above, and 79 (84.9%) had blood tests. The median age of patients was 44 (range, 21-78) years. The majority of women presented with menstrual disorders. Uterine pathology was detected on transvaginal scan in 42 (47.2%) cases. Adnexal pathology was detected in 12 (13.5%) of the patients. Endometrial biopsy detected three (4.5%) cases of endometrial atypia, and three (4.5%) cases of adenocarcinoma. A hemoglobin level of < 10 g/dL was detected in 3 (3.4%) patients. A single clinic visit was thought sufficient for 83 (89.2%) women. Medical therapy was started in 47 (50.5%) patients, 15 (16.3%) were brought back for inpatient diagnostic hysteroscopy, and nine (9.7%) were booked for operative endoscopy, while six (6.5%) had conventional surgery; the remainder were reassured. CONCLUSIONS: This study demonstrates that a 'one stop' management philosophy based on pelvic ultrasound is feasible. Our data suggest that diagnostic hysteroscopy can be decreased using this approach, and challenge the use of hysteroscopy as the first stage test. PMID- 11422976 TI - Two case reports of rare fetal tumors: an indication for surface rendering? AB - Fetal tumors are rare and can be difficult to diagnose in utero. This report describes one case of an extrarenal rhabdoid tumor and one case of a giant congenital melanocytic nevus. Both presented with moderate polyhydramnios and were prenatally detected at 31 weeks of gestation with two-dimensional ultrasound. The application of the surface mode of three-dimensional ultrasound improved the visualization of these tumors especially for the parents and the multidisciplinary team. Three-dimensional sonography proved to be a valuable addition to the prenatal armamentarium for the evaluation of these fetal tumors although it is not mandatory for their assessment and clinical management. PMID- 11422977 TI - Fetal anemia as a consequence of hemorrhage into an ovarian cyst. AB - Fetal ovarian cysts are usually benign in nature and resolve spontaneously. Complications such as torsion and rupture have been described previously. A case of presumptive spontaneous hemorrhage into a fetal ovarian cyst is presented. Serial ultrasound and Doppler assessments revealed the development of fetal anemia, which was managed by intrauterine blood transfusion. PMID- 11422978 TI - Prenatal diagnosis of type A1 brachydactyly. AB - Brachydactyly can occur as an isolated malformation or as part of numerous syndromes. Prenatal assessment of brachydactyly may be especially helpful in multiple anomaly syndromes associated with hand and/or finger anomalies. In isolated type A1 brachydactyly, which is an autosomal dominant disorder, all middle phalanges of the fingers and toes are affected. We present a fetus with type A1 brachydactyly inherited from the mother and grandmother. PMID- 11422979 TI - First-trimester prenatal diagnosis of a familial subtelomeric translocation. AB - A new fluorescent in situ hybridization (FISH) technique utilizes a complete set of telomeric probes to screen for deletions or rearrangements within the subtelomeric regions of all chromosomes on a single slide. Such cryptic chromosome rearrangements would otherwise remain undetected by standard cytogenetic analysis. In this case report, we describe the first-trimester prenatal diagnosis of an unbalanced rearrangement in a family where such a cryptic subtelomeric rearrangement is segregating. Interestingly the fetus was also noted to have an increased nuchal translucency at the time first-trimester chorionic villus sampling was performed and a FISH diagnosis made. The result was subsequently confirmed on fetal material obtained after elective termination of the pregnancy. We believe this to be the first report in the literature (as by Medline, December 1999) of a first-trimester prenatal diagnosis using such subtelomeric probes where confirmation by conventional cytogenetic analysis was not possible. PMID- 11422980 TI - Prenatal sonographic diagnosis of a twinning epigastric heteropagus. AB - Epigastric heteropagus is a rare type of conjoined twinning which results from an ischemic atrophy of one fetus at an early stage of gestation. We present what we believe to be the first case diagnosed antenatally at 22 weeks' gestation. The pelvis and lower limbs of the ischemic fetus (the parasite) were attached to the epigastrium of the well-developed fetus (the autosite), which had a small omphalocele. Antenatal sonography provided an accurate diagnosis, enabling unnecessary abortion to be avoided. PMID- 11422981 TI - Sonographic findings of the umbilical cord: implications for the risk of fetal chromosomal anomalies. AB - In this review we summarize current knowledge on sonographic findings of the umbilical cord and the risk they impose for chromosomal abnormalities of the fetus. A Medline search of the literature was performed and the pertinent English language literature was reviewed. Anatomical and Doppler abnormalities of the umbilical cord may be associated with an increased risk of chromosomal aberrations in the fetus. Therefore, level II prenatal sonography should also include a careful examination of the umbilical cord. PMID- 11422982 TI - Sirenomelia: early prenatal diagnosis with combined two- and three-dimensional sonography. PMID- 11422983 TI - Trisomy 7 following assisted conception treatment. PMID- 11422984 TI - Re: Is obsessional measurement taking worthwhile? PMID- 11422985 TI - Prenatal sonographic diagnosis of Pena-Shokeir syndrome type I. PMID- 11422987 TI - The feline glaucomas: 82 cases (1995-1999). AB - OBJECTIVE: To describe signalment, ophthalmic abnormalities, and response to treatment in cats with glaucoma. DESIGN: Retrospective study. ANIMALS: Eighty-two cats with 93 glaucomatous eyes. CRITERIA FOR INCLUSION: Medical records of all cats with glaucoma presented to the ophthalmology services at two referral specialty hospitals between 1995 and 1999 were reviewed. Cats were included if intraocular pressure (IOP) > 25 mmHg, if buphthalmos was present, or if a shallow anterior chamber was present and IOP > or = 5 mm Hg higher than the contralateral normal eye. RESULTS: Mean +/- SD age was 9.2 +/- 4.4 years. Thirty-one cats were female; 51 were male. Breeds included 69 domestic and 13 pure-bred cats. One eye was affected in 71 cats; both eyes in 11 cats. The most common presenting complaints and ophthalmic abnormalities were cataract, corneal edema, mydriasis, buphthalmos, cloudy eye, and blindness. Mean intraocular pressure in the affected eye was 36.4 +/- 14.7 mmHg. The glaucomas were believed to be secondary in 81 eyes, primary in five eyes, and undetermined in seven eyes. The goal of medical therapy was to maintain IOP in a comfortable range we presumed to be < 30 mm Hg in blind eyes, and to maintain IOP below 25 mmHg to preserve vision in visual eyes. Intraocular pressure was maintained in a comfortable range or normal range in 58% of all eyes using medical therapy alone. Surgery was performed in nine eyes after medical management failed. Sixty-seven eyes were blind, 21 were visual, and five had decreased vision at initial presentation. With treatment, vision was retained in nine eyes, deteriorated in five eyes, and was lost in three eyes. CONCLUSIONS: The glaucomas were most frequently secondary in cats and resulted in blindness prior to presentation in the majority of eyes in our study. Medical management controlled IOP in more than half of the eyes, and maintained vision in almost half of visual eyes. PMID- 11422988 TI - Clinical ocular toxoplasmosis in neonatal kittens. AB - OBJECTIVE: To describe ocular findings in kittens with congenital or early neonatal infection by Toxoplasma gondii and to determine if there are detectable differences in disease caused by three strains. ANIMALS STUDIED: Six adult female cats and the offspring from seven litters. METHODS: Four kittens from uninfected specific pathogen-free (SPF) queens and 21 kittens from SPF queens inoculated at various times late in gestation with Mozart, Maggie, or ME-49 strain of T. gondii were used. Ocular examinations were performed on queens prior to and after delivery, and on kittens weekly to bi-weekly for up to 27 weeks. Whole blood for serology was collected from all kittens at 5(1/2) to 8 weeks of age and again at 12 weeks of age or later. RESULTS: No kittens from noninfected queens developed ocular lesions or antibody to T. gondii. Three of the 24 kittens from infected queens died or were euthanized early in the study. Chorioretinitis was detected in 15 of 21 living kittens from infected queens. Two developed concurrent anterior uveitis that resolved within 1 week. Posterior segment lesions varied ophthalmoscopically between strains. Of 21 kittens from T. gondii-infected queens, six developed positive antibody titers to T. gondii during the study. All seropositive kittens were born to queens infected with Mozart strain of T. gondii. CONCLUSION: Results of this study suggest that ocular toxoplasmosis can occur without other evidence of clinical illness in kittens infected in utero or in the neonatal period, and that T. gondii strains may have varying degrees of ocular pathogenicity in cats. PMID- 11422989 TI - Lipogranulomatous conjunctivitis: clinical findings from 21 eyes in 13 cats. AB - OBJECTIVE: The clinical and, where available, histologic features of lesions fitting a previous description of lipogranulomatous conjunctivitis were reviewed retrospectively from a series of cats. ANIMALS STUDIES: Thirteen cats (21 eyes) with lesions consistent with a diagnosis of lipogranulomatous conjunctivitis were seen over a 4-year period in a referral practice in southern Australia. Affected cats ranged in age from 6 to 16 years (mean 11.2 +/- 3.6 years). RESULTS: Twelve cats were domestic shorthair or longhair cats and one was a British shorthair cat. Five cats were white, four were bicolor, two were orange tabby and two were dilute orange tabby. Lesions were always found in the palpebral conjunctiva, adjacent to the eyelid margin. They affected upper eyelids only in five cases, and upper plus lower eyelids in eight cases. Lesions were bilateral in eight cases. Lesions were excised in eight cases, with resolution of associated irritation and no signs of lesion recurrence during a 4- to 21-month follow-up period. SUMMARY: Histologic features of tissues excised from surgically treated cases were consistent with a previous report of lipogranulomatous conjunctivitis and with descriptions of chalazia. A possible role of actinic radiation in the etiopathogenesis of lesions is discussed. PMID- 11422990 TI - Enrofloxacin-associated retinal degeneration in cats. AB - OBJECTIVE: The objective of this study was to evaluate the possible relationship between the administration of parenteral and/or oral [corrected] enrofloxacin and the onset of acute retinal degeneration in cats. The animals studied included 17 cats that received systemic enrofloxacin and developed retinal degeneration soon thereafter. PROCEDURES: In this retrospective clinical study, cats that received parenteral and/or oral [corrected] enrofloxacin and developed acute blindness were identified. Parameters recorded included breed, age, sex, enrofloxacin dosage (daily dose and number of days administered), medical condition for which the antibiotic had been prescribed, ophthalmic signs, examination results, and the visual outcome. Fundus photographs were obtained in seven cats, and electroretinography was performed in five cats. Histopathology was performed on two eyes from one cat (case 1) that received enrofloxacin 5 months previously and developed retinal degeneration. RESULTS: All cats were the domestic shorthair breed; seven were females (one neutered) and ten were males (seven castrated). Ages ranged from 3 to 16 years old (mean +/- SD; 8.8 +/- 4.6 years). The medical disorders for which enrofloxacin was administered ranged from lymphoma and pancreatitis to otitis and dermatitis, and eight cats had urinary diseases. The daily and total dosage of enrofloxacin and number of days of administration were also highly variable. Presenting clinical signs were most often mydriasis and acute blindness. All cats had diffuse retinal degeneration as evidenced by increased tapetal reflectivity and retinal vascular attenuation. Absence of recordable electroretinographic responses suggested diffuse and extensive outer retinal disease. Vision returned in a few cats, but the retinal degeneration persisted or even progressed. Histopathology of two eyes revealed primarily outer retinal degeneration, with diffuse loss of the outer nuclear and photoreceptor layers, and hypertrophy and proliferation of the retinal pigment epithelium. CONCLUSION: Parenteral and/or oral [corrected] enrofloxacin is potentially retinotoxic in some cats, and may result in acute and diffuse retinal degeneration. Blindness often results, but some cats may regain vision. Practitioners should adhere closely to the manufacturer's current enrofloxacin dosage recommendation (5 mg/kg q 24 h), and continue clinical observations for this drug toxicity in cats. PMID- 11422991 TI - Corneoconjunctival transposition for the treatment of feline corneal sequestra: a retrospective study of 17 cases (1990-1998). AB - OBJECTIVE: To describe and evaluate the use of corneoconjunctival transposition as a surgical treatment for corneal sequestra in cats. ANIMALS STUDIED: Fifteen cats (17 eyes) of various breeds and ages with corneal sequestra treated surgically with corneoconjunctival transposition. PROCEDURE: Retrospective medical records study. RESULTS: Median age was 39 months. Eight patients were castrated males, five were spayed females, and two were intact females. The left eye was affected in eight cats, the right eye was affected in five cats, and both eyes were affected in two cats. Mean (+/- SD) sequestrum diameter was 4.4 +/- 2.4 mm (range 1-10 mm) and the sequestrum was located centrally in all cases. Sequestra occurred in the superficial stroma in 8/17 corneas, in the anterior one third of the stroma in 3/17 corneas, and in the middle one third of the stroma in 6/17 corneas. Median duration of clinical signs prior to presentation was 39 days (range 14-1095 days). Median time to healing, defined as the number of days from surgery to cessation of medical treatment, was 34 days (range 25-86 days). There were no recurrences and minimal scar formation. CONCLUSIONS: Corneoconjunctival transposition is a valid procedure for surgical treatment of corneal sequestra in cats. The procedure results in excellent cosmesis and functional vision with no recurrences. PMID- 11422992 TI - Phacoemulsification in a clouded leopard (Neofelis nebulosa). AB - Phacoemulsification and prophylactic retinopexy were performed on a 3-month-old hand-raised clouded leopard with a unilateral mature cataract. The same procedures were repeated 3 months later when a cataract developed in the fellow eye. Examination procedures, anesthetic protocols, and surgical techniques were similar to those utilized in domestic cats. The patient retains highly functional aphakic vision at the present age of 2.5 years. An inherited basis for the cataracts is proposed. Although the cub was hand raised, the cataracts were unlike nutritional cataracts reported previously in domestic cats and dogs. To the author's knowledge this is the first report of cataract formation and phacoemulsification in a clouded leopard. PMID- 11422993 TI - Feline leukemia virus detection in corneal tissues of cats by polymerase chain reaction and immunohistochemistry. AB - OBJECTIVES: To determine the presence of feline leukemia virus (FeLV) in the corneas of FeLV-infected cats. ANIMALS STUDIED: Thirty-four random source cats. PROCEDURES: Seventeen cats were found positive for FeLV serum p27 antigen by enzyme-linked immunosorbent assay (ELISA). Twelve ELISA positive cats were also positive on peripheral blood samples by immunofluorescent antibody (IFA) testing. Seventeen ELISA negative cats served as negative controls. Full thickness corneal specimens were collected from the left cornea of all cats and analyzed for FeLV proviral DNA and gp70 antigen by polymerase chain reaction (PCR) and immunohistochemical (IHC) testing, respectively. RESULTS: Eleven (64.7%) positive corneal PCR results were obtained from 17 ELISA positive cats. Of 12 cats that were both ELISA and IFA positive on peripheral blood, 10 (83.3%) had positive corneal PCR results. All corneal tissues from ELISA negative cats were PCR negative. IHC staining of corneal sections revealed the presence of FeLV gp70 in corneal tissues of nine (52.9%) ELISA positive cats. Of the 12 cats that were both ELISA and IFA positive on peripheral blood, eight (66.7%) had positive corneal IHC results. Positive IHC staining was localized to the corneal epithelium. Corneal tissues of all ELISA negative cats and all IFA negative cats were negative on IHC testing. CONCLUSIONS: FeLV antigens and proviral DNA are present in corneal tissues of some FeLV-infected cats. Screening corneal donors for FeLV infection is warranted. PMID- 11422994 TI - A comparison of corneal sensitivity between brachycephalic and Domestic Short haired cats. AB - OBJECTIVE: To compare sensitivity of the central and peripheral cornea of brachycephalic and Domestic Short-haired (DSH) cats. DESIGN: Prospective study. ANIMALS: Fifty DSH, thirteen Persian, and seven Himalayan cats. CRITERIA FOR INCLUSION: Healthy DSH, Persian, or Himalayan cats older than 6 months, with a normal ophthalmic examination and Sno-strip values greater than or equal to 5 mm wetting/60 s. MATERIALS AND METHODS: Corneal sensitivity was measured using a Cochet-Bonnet aesthesiometer. The cornea was touched using the tip of the monofilament, decreasing the fiber length in 5 mm increments until a blink reflex occurred. Corneal touch threshold was defined as the stimulus that elicited a corneal blink reflex greater than 50% of the time. RESULTS: Mean central and peripheral corneal touch threshold (CTT, +/- SD in gm/mm2) for the DSH cats were 1.79 +/- 2.33 and 5.01 +/- 5.07 in the right eye (OD), and 1.74 +/- 1.65 and 5.02 +/- 4.55 in the left eye (OS). Mean central and peripheral CTT +/- SD (gm/mm2) for the brachycephalic cats were 4.09 +/- 5.29 and 6.18 +/- 5.65 OD, and 3.18 +/- 3.75 and 7.66 +/- 6.24 OS. Statistical analysis of the data revealed a significantly higher CTT in the central (P = 0.019) and peripheral (P = 0.003) cornea of brachycephalic than DSH cats. When evaluated for gender, this difference persisted in female cats, but did not hold true for male cats. A significant difference in CTT was found between central and peripheral cornea within both groups of cats. CONCLUSIONS: The central cornea is less sensitive in brachycephalic than DSH cats. The central cornea is more sensitive than the peripheral cornea in both brachycephalic and DSH cats. PMID- 11422995 TI - Systemic hypertensive disease and the feline fundus. AB - The fundoscopic appearance and some of the histopathological findings of arterial hypertension in the cat are reviewed in relation to the anatomical and physiological features that place retinal function at particular risk when the eye is subjected to sustained increased arterial blood pressure. The fundus changes fall into three categories: hypertensive retinopathy, hypertensive choroidopathy and hypertensive optic neuropathy, and information from cases with confirmed arterial hypertensive disease is used to provide a basis for discussion and future investigation. PMID- 11422996 TI - Immunohistochemical studies of cone photoreceptors and cells of the inner retina in feline rod-cone degeneration. AB - Previous studies using electroretinography and immunohistochemistry have shown normal cone function and structure in early stages of hereditary rod-cone degeneration of Abyssinian cats. To further investigate the cone photoreceptors and the inner retina of dystrophic cats, antibodies against green- and blue sensitive cones and specific cell types of inner retina were used in seven cats with the recessively inherited rod-cone degeneration, and three normal European short-haired cats. There was a reduction in number of both types of cones early in the disease. Changes at early stages of disease also occurred among horizontal cells in which there was an extension and a thickening of their lateral processes. The regular configuration of bipolar cells was changed in the more advanced stages of disease and their apical dendrites were lost. Abnormalities were not observed in the amacrine cells and in the ganglion cell layer in any of the present cases. This study shows that the cone system is morphologically abnormal in young cats at an earlier stage of disease than previously shown. The present findings also support the assumption that the inner retina is largely preserved throughout the disease process. PMID- 11422997 TI - The use of porcine small intestinal submucosa in ten cases of feline corneal disease. AB - Porcine small intestinal submucosa (SIS) was used as a novel graft material in the management of 10 cases of feline corneal disease. Five cases had stromal ulceration associated with trauma, ocular foreign body and/or suspected infection and required a grafting procedure. Five cases had feline sequestra that were managed by a keratectomy prior to placement of SIS as a graft material. Eight eyes healed with minimal corneal scarring with a very good cosmetic and visual result. One eye with continued aqueous leakage in the immediate postoperative period required a conjunctival pedicle graft to reinforce the SIS graft site. One eye required enucleation 48 h following grafting due to progressive keratomalacia but the SIS material remained intact. PMID- 11422998 TI - Acute vision loss after general anesthesia in a cat. AB - A 6-year-old, male castrated, domestic shorthair cat presented with a 2-week history of blindness following general anesthesia. Behavioral changes, lethargy, inappetence and neurological deficits were also noted. Ophthalmic evaluation revealed blindness, normal pupillary light response and dazzle response, but no other abnormalities. Serodiagnostic testing for common infectious agents was negative and a cerebrospinal fluid analysis was normal. History and postmortem examination following euthanasia revealed cerebrocortical necrosis most consistent with anesthesia related hypoxia. PMID- 11422999 TI - A modification of the Mustarde technique for the surgical repair of a large feline eyelid coloboma. AB - A one-year-old male domestic short haired cat was referred because of an area of keratitis that had developed adjacent to a particularly large unilateral eyelid coloboma. The defect was repaired using an undescribed three-stage surgical technique, which is essentially a combination of existing techniques and consists of using the lower lid to re-construct the upper lid and replacing the resultant lower lid defect with a full thickness graft from the upper lip. This technique is described and the results illustrated by photographs. A good functional and cosmetic result was achieved. PMID- 11423000 TI - Persistent hyperplastic tunica vasculosa lentis and persistent hyperplastic primary vitreous (PHTVL/PHPV) in two cats. AB - Two domestic shorthair cats (6 and 9 months old) were presented for examination of ocular opacities. One cat had bilateral persistent pupillary membranes, unilaterally accompanied by persistent hyperplastic tunica vasculosa lentis, persistent hyperplastic primary vitreous (PHTVL/PHPV), and cataract. The second cat had bilateral PHTVL/PHPV with dense white plaques in the posterior lens capsule and subcapsular cortex. PMID- 11423001 TI - Genomic organization and alternative splicing of the human and mouse RPTPrho genes. AB - BACKGROUND: Receptor protein tyrosine phosphatase rho (RPTPrho, gene symbol PTPRT) is a member of the type IIB RPTP family. These transmembrane molecules have been linked to signal transduction, cell adhesion and neurite extension. The extracellular segment contains MAM, Ig-like and fibronectin type III domains, and the intracellular segment contains two phosphatase domains. The human RPTPrho gene is located on chromosome 20q12-13.1, and the mouse gene is located on a syntenic region of chromosome 2. RPTPrho expression is restricted to the central nervous system. RESULTS: The cloning of the mouse cDNA, identification of alternatively spliced exons, detection of an 8 kb 3'-UTR, and the genomic organization of human and mouse RPTPrho genes are described. The two genes are comprised of at least 33 exons. Both RPTPrho genes span over 1 Mbp and are the largest RPTP genes characterized. Exons encoding the extracellular segment through the intracellular juxtamembrane 'wedge' region are widely spaced, with introns ranging from 9.7 to 303.7 kb. In contrast, exons encoding the two phosphatase domains are more tightly clustered, with 15 exons spanning approximately 60 kb, and introns ranging in size from 0.6 kb to 13.1 kb. Phase 0 introns predominate in the intracellular, and phase 1 in the extracellular segment. CONCLUSIONS: We report the first genomic characterization of a RPTP type IIB gene. Alternatively spliced variants may result in different RPTPrho isoforms. Our findings suggest that RPTPrho extracellular and intracellular segments originated as separate modular proteins that fused into a single transmembrane molecule during a later evolutionary period. PMID- 11423002 TI - Determinants of patient recruitment in a multicenter clinical trials group: trends, seasonality and the effect of large studies. AB - BACKGROUND: We examined whether quarterly patient enrollment in a large multicenter clinical trials group could be modeled in terms of predictors including time parameters (such as long-term trends and seasonality), the effect of large trials and the number of new studies launched each quarter. We used the database of all clinical studies launched by the AIDS Clinical Trials Group (ACTG) between October 1986 and November 1999. Analyses were performed in two datasets: one included all studies and substudies (n = 475, total enrollment 69,992 patients) and the other included only main studies (n = 352, total enrollment 57,563 patients). RESULTS: Enrollment differed across different months of the year with peaks in spring and late fall. Enrollment accelerated over time (+27 patients per quarter for all studies and +16 patients per quarter for the main studies, p < 0.001) and was affected by the performance of large studies with target sample size > 1,000 (p < 0.001). These relationships remained significant in multivariate autoregressive modeling. A time series based on enrollment during the first 32 quarters could forecast adequately the remaining 21 quarters. CONCLUSIONS: The fate and popularity of large trials may determine the overall recruitment of multicenter groups. Modeling of enrollment rates may be used to comprehend long-term patterns and to perform future strategic planning. PMID- 11423003 TI - Location, location, location. PMID- 11423004 TI - Chemical genomics: what will it take and who gets to play? AB - Chemical genomics requires continued advances in combinatorial chemistry, protein biochemistry, miniaturization, automation, and global profiling technology. Although innovation in each of these areas can come from individual academic labs, it will require large, well-funded centers to integrate these components and freely distribute both data and reagents. PMID- 11423005 TI - The many uses of a genome sequence. PMID- 11423006 TI - Where are they going? Directed cell movement in morphogenesis. PMID- 11423007 TI - The human olfactory receptor repertoire. AB - BACKGROUND: The mammalian olfactory apparatus is able to recognize and distinguish thousands of structurally diverse volatile chemicals. This chemosensory function is mediated by a very large family of seven-transmembrane olfactory (odorant) receptors encoded by approximately 1,000 genes, the majority of which are believed to be pseudogenes in humans. RESULTS: The strategy of our sequence database mining for full-length, functional candidate odorant receptor genes was based on the high overall sequence similarity and presence of a number of conserved sequence motifs in all known mammalian odorant receptors as well as the absence of introns in their coding sequences. We report here the identification and physical cloning of 347 putative human full-length odorant receptor genes. Comparative sequence analysis of the predicted gene products allowed us to identify and define a number of consensus sequence motifs and structural features of this vast family of receptors. A new nomenclature for human odorant receptors based on their chromosomal localization and phylogenetic analysis is proposed. We believe that these sequences represent the essentially complete repertoire of functional human odorant receptors. CONCLUSIONS: The identification and cloning of all functional human odorant receptor genes is an important initial step in understanding receptor-ligand specificity and combinatorial encoding of odorant stimuli in human olfaction. PMID- 11423008 TI - Extracting biological information from DNA arrays: an unexpected link between arginine and methionine metabolism in Bacillus subtilis. AB - BACKGROUND: In global gene expression profiling experiments, variation in the expression of genes of interest can often be hidden by general noise. To determine how biologically significant variation can be distinguished under such conditions we have analyzed the differences in gene expression when Bacillus subtilis is grown either on methionine or on methylthioribose as sulfur source. RESULTS: An unexpected link between arginine metabolism and sulfur metabolism was discovered, enabling us to identify a high-affinity arginine transport system encoded by the yqiXYZ genes. In addition, we tentatively identified a methionine/methionine sulfoxide transport system which is encoded by the operon ytmIJKLMhisP and is presumably used in the degradation of methionine sulfoxide to methane sulfonate for sulfur recycling. Experimental parameters resulting in systematic biases in gene expression were also uncovered. In particular, we found that the late competence operons comE, comF and comG were associated with subtle variations in growth conditions. CONCLUSIONS: Using variance analysis it is possible to distinguish between systematic biases and relevant gene-expression variation in transcriptome experiments. Co-variation of metabolic gene expression pathways was thus uncovered linking nitrogen and sulfur metabolism in B. subtilis. PMID- 11423009 TI - Evolution of gene order conservation in prokaryotes. AB - BACKGROUND: As more complete genomes are sequenced, conservation of gene order between different organisms is emerging as an informative property of the genomes. Conservation of gene order has been used for predicting function and functional interactions of proteins, as well as for studying the evolutionary relationships between genomes. The reasons for the maintenance of gene order are still not well understood, as the organization of the prokaryote genome into operons and lateral gene transfer cannot possibly account for all the instances of conservation found. Comprehensive studies of gene order are one way of elucidating the nature of these maintaining forces. RESULTS: Gene order is extensively conserved between closely related species, but rapidly becomes less conserved among more distantly related organisms, probably in a cooperative fashion. This trend could be universal in prokaryotic genomes, as archaeal genomes are likely to behave similarly to bacterial genomes. Gene order conservation could therefore be used as a valid phylogenetic measure to study relationships between species. Even between very distant species, remnants of gene order conservation exist in the form of highly conserved clusters of genes. This suggests the existence of selective processes that maintain the organization of these regions. Because the clusters often span more than one operon, common regulation probably cannot be invoked as the cause of the maintenance of gene order. CONCLUSIONS: Gene order conservation is a genomic measure that can be useful for studying relationships between prokaryotes and the evolutionary forces shaping their genomes. Gene organization is extensively conserved in some genomic regions, and further studies are needed to elucidate the reason for this conservation. PMID- 11423010 TI - Survey of human mitochondrial diseases using new genomic/proteomic tools. AB - BACKGROUND: We have constructed Bayesian prior-based, amino-acid sequence profiles for the complete yeast mitochondrial proteome and used them to develop methods for identifying and characterizing the context of protein mutations that give rise to human mitochondrial diseases. (Bayesian priors are conditional probabilities that allow the estimation of the likelihood of an event - such as an amino-acid substitution - on the basis of prior occurrences of similar events.) Because these profiles can assemble sets of taxonomically very diverse homologs, they enable identification of the structurally and/or functionally most critical sites in the proteins on the basis of the degree of sequence conservation. These profiles can also find distant homologs with determined three dimensional structures that aid in the interpretation of effects of missense mutations. RESULTS: This survey reports such an analysis for 15 missense mutations, one insertion and three deletions involved in Leber's hereditary optic neuropathy, Leigh syndrome, mitochondrial neurogastrointestinal encephalomyopathy, Mohr-Tranebjaerg syndrome, iron-storage disorders related to Friedreich's ataxia, and hereditary spastic paraplegia. We present structural correlations for seven of the mutations. CONCLUSIONS: Of the 19 mutations analyzed, 14 involved changes in very highly conserved parts of the affected proteins. Five out of seven structural correlations provided reasonable explanations for the malfunctions. As additional genetic and structural data become available, this methodology can be extended. It has the potential for assisting in identifying new disease-related genes. Furthermore, profiles with structural homologs can generate mechanistic hypotheses concerning the underlying biochemical processes - and why they break down as a result of the mutations. PMID- 11423011 TI - Evolution of mammalian genome organization inferred from comparative gene mapping. AB - Comparative genome analyses, including chromosome painting in over 40 diverse mammalian species, ordered gene maps from several representatives of different mammalian and vertebrate orders, and large-scale sequencing of the human and mouse genomes are beginning to provide insight into the rates and patterns of chromosomal evolution on a whole-genome scale, as well as into the forces that have sculpted the genomes of extant mammalian species. PMID- 11423012 TI - Endogenous retroviruses in the human genome sequence. AB - The human genome contains many endogenous retroviral sequences, and these have been suggested to play important roles in a number of physiological and pathological processes. Can the draft human genome sequences help us to define the role of these elements more closely? PMID- 11423013 TI - The origin and early evolution of mitochondria. AB - Complete sequences of numerous mitochondrial, many prokaryotic, and several nuclear genomes are now available. These data confirm that the mitochondrial genome originated from a eubacterial (specifically alpha-proteobacterial) ancestor but raise questions about the evolutionary antecedents of the mitochondrial proteome. PMID- 11423014 TI - Bases and spaces: resources on the web for accessing the draft human genome - II after publication of the draft. AB - The volume of human genome sequence and the variety of web-based tools to access it continue to grow at an impressive rate, but a working knowledge of certain key resources can be sufficient to get the most from your genome. This article provides an update to Genome Biology 2000, 1(4):reviews2001.1-2001.5. PMID- 11423016 TI - Current status of injectable agents for female stress urinary incontinence. AB - The purpose of this review is to summarize the results of reports of injectable agents for the treatment of female urinary stress incontinence. Five agents were reviewed: collagen, Teflon, autologous fat, silicone microparticles, and silicone microballoons. Collagen was the most frequently reported agent and yielded short term cure and improved rates of 74%-100%. This deteriorated to approximately 57% with longer term follow-up. Teflon has similarly lower longer term than short term success rates, at 33%-76%. The reported local complications and the potential for particle migration have resulted in its lack of widespread acceptance. Autologous fat has yielded the lowest success rate. Longer term success in a small number of silicone microparticle articles was similar to the other injectables at approximately 60%. Early success with silicone microballoons was 70%. The technique is generally free of major morbidity. The indication for injectables is intrinsic sphincter deficiency but hypermobility is not a contraindication. Long-term durability, cost effectiveness, and some safety issues still have to be addressed by further clinical trials. PMID- 11423015 TI - Importin-beta-like nuclear transport receptors. AB - SUMMARY: In recent years, our understanding of macromolecular transport processes across the nuclear envelope has grown dramatically, and a large number of soluble transport receptors mediating either nuclear import or nuclear export have been identified. Most of these receptors belong to one large family of proteins, all of which share homology with the protein import receptor importin beta (also named karyopherin beta). Members of this family have been classified as importins or exportins on the basis of the direction they carry their cargo. To date, the family includes 14 members in the yeast Saccharomyces cerevisiae and at least 22 members in humans. Importins and exportins are regulated by the small GTPase Ran, which is thought to be highly enriched in the nucleus in its GTP-bound form. Importins recognize their substrates in the cytoplasm and transport them through nuclear pores into the nucleus. In the nucleoplasm, RanGTP binds to importins, inducing the release of import cargoes. In contrast, exportins interact with their substrates only in the nucleus in the presence of RanGTP and release them after GTP hydrolysis in the cytoplasm, causing disassembly of the export complex. Thus, common features of all importin-beta-like transport factors are their ability to shuttle between the nucleus and the cytoplasm, their interaction with RanGTP as well as their ability to recognize specific transport substrates. PMID- 11423017 TI - The value of cystoscopy, prostate biopsy and frozen-section urethral biopsy prior to orthotopic neobladder substitution. AB - INTRODUCTION: The traditional criteria for selecting suitability of male patients with bladder cancer for othtopic neobladder substitution includes: patient motivation, negative transurethral (TUR) prostate biopsy prior to cystectomy, and/or tumor-free biopsy of the urethral margin at the time of cystectomy. In this report we evaluate the value of these preoperative and intraoperative biopsies in the cystoscopically normal prostatic urethra. METHODS: During a 5 year period, cystectomies were performed on 54 men and 13 women with invasive bladder cancer. Prior to this procedure, all of the men had TUR biopsy of the prostate. Forty men with cystoscopically normal prostatic fossa had negative prostatic biopsies, and. had orthotopic neobladder substitution. RESULTS: Pathological examination showed that 3 out of these 40 men had transitional cell carcinoma (TCC) involving the prostate. In all 40 patients, the prostatic apical resection margin was negative. To date, none of the patients developed urethro neobladder recurrence. CONCLUSION: TUR biopsy of the prostatic urethra prior to cystectomy, or biopsy of the prostatic urethral margin at the time of cystectomy in patients with cystoscopically normal, tumor free prostatic urethra has limited value in selecting candidates for orthotopic neobladder substitution. PMID- 11423018 TI - Management of penile toilet seat injury--report of two cases. AB - Blunt trauma to the penis is an uncommon injury in young children. We present two cases of blunt penile trauma secondary to mechanical compression from a toilet seat. PMID- 11423019 TI - Metastatic renal cell carcinoma presenting as an oral tumor. AB - Metastatic cancer presenting as an oral lesion is exceedingly uncommon. To the best of our knowledge this is the first reported instance of renal cell carcinoma presenting initially as an oral lesion. PMID- 11423020 TI - Laparoscopic wedge resection of a renal mass in a solitary kidney. AB - Laparoscopic radical nephrectomy has gained acceptance as a viable means of managing renal neoplasms. Partial nephrectomy has traditionally been performed through a flank or transabdominal incision, with its attendant morbidity and required hospitalization. Laparoscopic renal-sparing surgery may be more technically demanding, but remains beneficial to the patient if performed with adherence to oncological principles. We present a case of laparoscopic renal sparing surgery in a patient with a renal mass in a solitary kidney. We discuss the technical and oncological challenges of this approach. PMID- 11423021 TI - [Health and immigration; talking about AIDS]. PMID- 11423022 TI - [Gaceta and the net]. PMID- 11423023 TI - [HIV seroprevalence and exposure categories among adolescents tested at a sexually transmitted clinic diseases. Madrid, 1986-2000]. AB - OBJECTIVE: To describe the time-trend in exposure categories and HIV seroprevalence among adolescents who underwent to voluntary testing in the period 1986-2000. METHODS: This study covered all adolescents, aged 13 to 19 years, at their first test for HIV in a sexually transmitted disease clinic in Madrid. Gender, age and HIV risk behaviours were collected. HIV diagnosis relies on ELISA test and Western blot confirmation. Time trends in HIV seroprevalence and exposure categories were analysed. RESULTS: A total of 1327 adolescents, 52% women and 22% under 18 years, were studied. The annual number of adolescents remained through the time, but injecting drug users (IDU) and IDU partners declined and female sex workers rose. 108 adolescents were diagnosed with HIV infection -71% were IDU-. HIV seroprevalence was 8.1% -31.3% in IDU-. It declined from 18.2% in 1986 to 1.5% in 1995, and after then it held steady under 4%. This decline involved several risk categories and was statistically significant in homo/bisexual men and female sex workers. The logistic regression analysis, adjusting for changes in exposure categories, showed an annual reduction in HIV seroprevalence (OR = 0.87; 95% CI, 0.81-0.94). CONCLUSIONS: HIV seroprevalence has decreased due to the fall of new young IDU and the decrease of seroprevalence within several exposure categories. HIV infections and risk behaviours continue happened among adolescents. PMID- 11423024 TI - [Changes in condom use among a cohort of prostitutes]. AB - OBJECTIVE: To describe changes in condom use among prostitutes enrolled in a prospective cohort study in Alicante (Spain), and to identify those factors associated with changes in this behaviour. METHODS: Data from this study come from a prospective cohort study of women working at prostitution enrolled in a center for AIDS preventive at Alicante from 1986 to 1996. The proportion of condom use by prostitutes with their clients was recorded during the first visit and the last one, and changes in condom use were estimated as the difference between them. Two logistic regression analyses were done to identify those factors associated with positive or negative changes in condom use. RESULTS: A statistically significant increase in the prevalence of condom use was observed between the first, 69,2%, and the last visit, 83,9% (p < 0,001). Prostitutes working private increased the condom use more than five time than those working in clubs or at the street. The higher the follow-up time the higher the prevalence of condom user, OR > 6 years = 7,6 (95% CI: 2,2-26,0). No variable was associated with a negative change in condom use. CONCLUSION: This study shows an overall increase of condom use in a cohort of prostitutes after 10 years of follow-up. Relapse in risk behavior has been infrequent. In the future, preventive strategies to increase the use of condoms among prostitutes should be adapted to the new forms of sexual commerce. PMID- 11423025 TI - [Health self-perception in men and women among the elderly]. AB - OBJECTIVES: To analyze the differences in health self-perception between men and women in the later stages of life and to assess their association with sociodemographic and health variables. METHODS: Data on 1,459 men and 1,993 women aged 60 or older from the 1994 Catalan Health Survey, were collected and an analysis of health self-perception according to age, gender, social class, reporting of chronic disease and handicaps was performed. Multivariate logistic regression analysis was used, taking into account the survey design. RESULTS: Of the women aged 60 and older, 57.3% reported poor health compared with 43.6% of men of the same age. The proportion of women with one or more handicaps was also greater (41.2%) than that of the men (28.7%), as was the case with chronic disease (92.2% in women and 85.6% in men). The multivariate model also revealed that health self-perception was poorer among women than among men. Other explanatory factors were the number of chronic diseases, having handicaps, and being a member of a lower social class. Age interacted with the number of chronic diseases to mitigate the effect of chronic diseases on perceived health status. CONCLUSIONS: In the elderly population, poor health self-perception was greater among women, even when other significant explanatory variable (social class, age, handicaps and chronic disease) were adjusted for. The most important explanatory factors in health self-perception were having chronic disease and/or handicaps. The impact of chronic disease on poor health self-perception decreased in older age groups. PMID- 11423026 TI - [Documented immunization coverage in second-year primary schoolchildren from the province of Valladolid]. AB - OBJECTIVE: To estimate the documented immunization coverage and degree to which schoolchildren's vaccination cards are kept up to date. METHODS: Transversal descriptive study of children in the second year of primary education in the province of Valladolid during the 1999-2000 academic year. The sample consisted of 698 children with a participation rate of 82%. The percentage of children whose immunization schedule was up to date in accordance with their age, that of children with additional vaccinations and the accuracy of the vaccination cards was quantified. Information was collected through a questionnaire on vaccination cards or, for children without one, from their medical history. RESULTS: The documented immunization coverage was 99.3% (95% CI: 98.6-99.9) for the first three dose of diphtheria toxoid, tetanus toxoid and pertussis vaccine and poliomyelitis vaccine, 98.9% (95% CI: 97.7-99.5) for the measles, mumps and rubella vaccine and 95% (95% CI: 93.4-96.7) for all doses up to the age of six. In addition, 7.2% (95% CI: 5.3-9.2) were immunized against hepatitis B, 14.4% (95% CI: 11.6-17) against Haemophilus influenzae type B and 90.3% (95% CI: 88 92.5) against meningococal A + C. A total of 84.4% of vaccination cards were correctly filled in (95% CI: 81.7-87.2). CONCLUSIONS: Systematic immunization coverage in the schoolchildren was high. Because of their accuracy, vaccination cards were a useful tool for determining immunization coverage. Both the accuracy of the vaccination card and the incidence of non-systematic immunization were higher in urban areas. PMID- 11423027 TI - [Characteristics of pregnant women and routine antenatal care in Barcelona, 1994 1999]. AB - OBJECTIVE: To analyse socio-demographic and pregnancy-control aspects of pregnant women residents in Barcelona city and their evolution during the period 1994-1999 using the controls of the Barcelona Birth Defects Registry. METHODS: The information was collected by mean of an interview to the mother and from hospital records. The Mantel-Haenszel method for lineal association was used to analyse trends. A chi-squared test for proportions was used to compare pregnancy-control variables between public and private centers. RESULTS: Data on 1,337 pregnant women were obtained. An increasing proportion of women older than 34 years is observed, from 19% in 1994-95 to 25% in 1998-99. An increase in the social class and a decrease of housewives is noticed. 40% of pregnancies were not planned and half of these finished in induced abortion. These proportions are higher in less than 25 and more than 39 years old mothers. 97% had their first obstetrical visit during the first trimester, almost all had at least an obstetrical ultrasound with a mean of 5.2, although 25% of the mothers did not undergo an ecography during the 5th month. An increase of invasive procedures and caesarean sections (C-section) is shown, with 33% of C-sections in the 98-99 period. The medical control of pregnancies is higher in mothers delivering in private centers, which are 55% of the total. CONCLUSIONS: There is a high lack of pregnancy planning, as well as an increasing pregnancy medicalization, with positive (first trimester visit) and debatable aspects (obstetrical ultrasound, caesarean section and invasive procedures). PMID- 11423028 TI - [Priority setting, public opinion and health care system in Spain]. AB - OBJECTIVES: To examine the determinants of attitudes towards some "general criteria" guiding the financing, provision and satisfaction of the Spanish health system. First we examine the degree of acceptance of a publicly funded health system such as the use of an intergenerational equity criteria for health care rationing based on patient's age. Second, employing the same sample we analyse the determinants of citizen's satisfaction with the health system in order to identify the profile that defines attitudes of Spanish population to their health system. METHODOLOGY: We undertake a quantitative analysis of the public opinion survey Eurobarometer 49.1 (1998) for a subsample of the Spanish population. The Eurobarometer is a periodical public opinion survey representative of European Union (EU) citizens. Due to the categorical nature of individual responses to public opinion surveys, the model estimated is an ordered probit. The explanatory variables used refer to socio-economic status and political attitudes. RESULTS: There appears to be a consensus on the criteria that public sector should go a way forward from what the public envisages as "essential health care". 73.5% of the populations rejects a libertarian criteria that sustains that individuals are self responsible for funding non-essential health care. This attitude is especially supported by male with leftists political tendencies and high education achievement. The use of age-related criteria to ration health care (fair innings) is rejected by a 81.5% of the population. However, we find that self interest is the main criteria guiding this attitude since elderly and middle and high income individuals tend reject the use of this criteria more than other groups. Satisfaction with the Spanish health system is higher than other southern EU countries, as Italy and Greece but still far from the levels achieved by Scandinavian and northern EU countries. Political attitudes, age and socio economic status are positively associated with a higher satisfaction. CONCLUSIONS: Health systems reforms that significantly reduce the collective funding of health systems would not be accepted by the majority of the population. As it happens in other EU countries, attitudes on the financing and provision of health care are influenced by political attitudes. Health reforms reducing the extent of health care funding would be rejected by the population. The use of and age-related criteria for health care rationing would be envisaged as discriminatory against the elderly. Health system satisfaction is in an intermediate position and its sensitive to demographic and socio-economic composition of the Spanish population, still far from the levels achieved by Scandinavian and northern EU countries. This results show a particular general criteria when evaluating health systems key elements, and may be expected to vary when applied to the concrete decision making scenario. Finally, it should be noted that quantitative analysis of general surveys is subject to large limitations. Thus, caution should be posed when interpreting these results, always should be seen as complementary of other studies using alternative methodologies (those using qualitative and experimental methodologies). PMID- 11423029 TI - [Management of research in healthcare centers. An exploration through nominal group dynamics]. AB - OBJECTIVES: To determine the opinions of hospital and other research organizations managers concerning three questions: 1) the desirability of healthcare centers' managing biomedical research as a specific product; 2) the characteristics that define a "research management" culture as opposed to the current "administration of research" culture and 3) the management instruments needed to implement a "research management" culture. MATERIALS AND METHODS: A meeting was held with 14 experts from healthcare centers or research organizations from Barcelona, Madrid and Valencia to discuss the three questions and evaluate the relative importance of items comprising the last two with a score system from 1 (little relevance) to 9 (extremely relevant). RESULTS: The group was in favor of healthcare centers' managing research as a specific and differentiated product. Keys to achieving a management culture (items scoring higher than 7 with the above system) included the development of a culture to evaluate work and external auditing, transversal support and sharing infrastructures and intellectual capital, specific accounting, unified management, prioritization of research lines and capacity for cooperation, and strategic alliances between centers. Management instruments deemed essential (items scoring 7 or higher with the above system) were: support to research foundations or other organizational formulae to ensure autonomy, specific budgetary control, development of support structures and contractual formulae to support autonomy, creativity and researchers' accountability. CONCLUSIONS: Research conducted in healthcare centers should be managed just as any other product derived from the center's activities. Key points to success are prioritization, evaluation and clear assignation of responsibilities, which requires organizational structures with greater flexibility and specific information systems. PMID- 11423030 TI - [Authorship and data reporting according to gender in four Spanish biomedical journals]. AB - AIMS: To describe authorship according to gender in the articles published in four Spanish scientific journals of general medicine and public health, and to analyse the type of presentation of the results in relation to the sex of the subjects of the study. METHODS: We examined 423 articles from four Spanish scientific journals published during 1998. RESULTS: 70.7% of the articles revised had a man as the first author. The sex difference was significant in the editorials (12.4% of the women vs. 29.8% of the men, p < 0.05). Stratification of the results by sex was presented in 167 articles and in 85.6% of them the information about men. DISCUSSION: Gender inequalities in the publication of scientific works in the field of general medicine and public health are found. PMID- 11423031 TI - [Diagnostic agreement between primary and hospital care in the assessment of basal cell carcinoma]. AB - OBJECTIVE: To evaluate the diagnostic agreement between primary and hospital care. METHODS: Cross-sectional study. 491 patients who sought primary care for symptoms of suspected basal cell carcinoma and were referred to a dermatologist. The Kappa index was used to evaluate diagnostic concordance between primary and hospital care. RESULTS: The more frequent site of diagnosis was face (73.9%). The most frequent differential diagnosis in primary care was nevi (5.9%) while that in specialist care was actinic keratoses (2.2%). The diagnostic agreement was very low (kappa = 0.071; 95% IC = 0.0-0.15). CONCLUSIONS: The degree of agreement between primary and specialist care was low. PMID- 11423032 TI - [Public policy analysis]. AB - This article presents to public health professionals concepts and perspectives from political science relevant for creating a healthier public policy. Currently, there is no uniform vision of what constitutes public interest and the decisions of public administrations tend to be based on compromise. In public debate, what is paramount is the capacity to persuade. From the perspective of public policy analysis, the crucial issue is definition: the final decision depends on the definition of the problem that has emerged triumphant in the public debate among competing actors with different definitions of the problem. From a policy analysis perspective, the problems entering the agenda of public administration does not necessarily correspond to their severity, as competing actors try to impose their point of view. Because of its historical evolution, the Spanish political system has specific traits. The relatively weak democratic tradition tends to make the decision process less visibles, with strong technocratic elements and weaker social articulation. Both the juridical tradition and liberal rhetoric portray lobbying as contrary to public interest, when in fact it is constantly performed by powerful vested interest groups, through both personal contacts and economic connections. Regulatory policies, with concentrated costs and diffuse benefits, seem to be moving from Spain to the European Union. To promote healthier public policies, the development of civil society initiatives and the building of coalitions will play an increasingly greater role in the future. PMID- 11423033 TI - [Smoking prevention as a political issue]. AB - The causes of many health problems such as smoking are beyond individual action. To change them requires the development of interventions dealing with the social factors that cause them, interventions of a political nature. In this paper we review for Spain public policies influencing smoking, with an emphasis on those with greater potential impact (fiscal policies to increase price, and those controlling the promotion and publicity of tobacco products), as well as the role of different government levels in regulation (local, regional central and European). The role of the Central government and of the European Union are crucial today in Spain. Some aspects influencing the adoption of smoking prevention policies (problem definition, private interests, and ideology) are also analyzed, as well as the need for a strategic coalition to promote them. PMID- 11423034 TI - [Estimate of daily cases of influenza from the cases notified to the Reportable Disease System: usefulness in time series studies]. AB - In time series analyses assessing the relationship between risk factors and respiratory diseases or mortality, influenza incidence is a potential cofounder and, therefore, must be controlled. Available influenza data come from weekly compulsory notifiable disease (EDO system). Furthermore, its graphical distribution, suggests that information may be underestimated in holiday periods. We have applied a procedure for estimate daily influenza series from the weekly cases, using less, a non-parametric local fit. Findings show that smoothing could avoid stepping and sudden peaks in the original series. PMID- 11423035 TI - [Exploring the National Cancer Institute's web site]. PMID- 11423038 TI - [Should we chose the cheapest, essentially similar drug?]. PMID- 11423040 TI - [Neonatal hypotonia]. PMID- 11423041 TI - [Neonatal hypotonia of muscular origin: analysis of 50 cases]. AB - BACKGROUND: Neonatal hypotonia is a common request for neurological consultation. The aim of this study is to describe the main clinical features of muscular hypotonia in newborns at the light of the histopathological findings. METHODS: We reviewed 50 medical records of hypotonic neonates with abnormal muscular biopsy. In all of the cases, the serum concentration of creatine kinase was determined and biopsies were examined by a qualified neuropathologist. RESULTS: The most frequent muscular cause of neonatal hypotonia was specific congenital myopathies (23 cases), followed by congenital muscular dystrophy (15 cases), congenital myotonic dystrophy (eight cases) and metabolic myopathies (four cases). The most common specific congenital myopathy was fiber type disproportion (10 cases). The association with joint contractures and the involvement of respiratory muscles were frequent; respiratory complications were the first cause of death. CONCLUSIONS: Muscular biopsy is required for the definitive diagnosis of specific congenital myopathies, congenital muscular dystrophy and metabolic myopathies. In congenital myotonic dystrophy, the mother is almost always affected; neonates with specific congenital myopathies and congenital muscular dystrophy can be very similar to those with congenital myotonic dystrophy; the examination of the mother, specially a careful search for myotonia, is the best diagnostic clue; if there are signs of myotonia, the diagnosis can be made by molecular genetic study. PMID- 11423042 TI - [Five by five test. A brief instrument for the detection of cognitive impediment in clinical settings]. AB - BACKGROUND AND OBJECTIVE: Many psychological tests for the detection of cognitive impediment and dementia are inadequate by being too prolonged, inconsistent or with poor specificity or sensibility. It is presented a brief test of evocation of proper names by classes (TIG) and their properties psicometrics and utility are compared with the Mini-mental State Examination (MEM). SUBJECTS AND METHODS: We selected 196 patients from a neurological clinic, with suggestive symptoms of cognitive impediment. We excluded individuals with focal or chronic cerebral disorders, sensorial disturbances, severe psychiatric diseases and toxicity by drugs. Patients were by grouped the Clinical Dementia Rating scale and they were evaluated neuropsychologically. The internal consistency of TIG was examined and a cross-validation procedure was performed to test for the predictive validity. Likewise, the convergent validity was tested against the scale of dementia of Blessed and the MEM. Finally the diagnostic efficiency of both tests, the effects of the demographics variables and the clinical utility were analyzed. We used non parametrics statistics, analysis of sensibility upon curves ROC and analysis of the post-test probabilities. RESULTS: The TIG has adequate internal consistency and validity. The diagnostic efficiency was high (area under the curve ROC 0,939 [IC 95%: 0,908-0,971], p < 0,0001). The introduction of the demographics covariates in a logistic model wisely improved the diagnostic efficiency. The TIG has some better fulfillment than the MEM. Neither the MEM nor the TIG are tests suitable for communitary screening of cognitive defect. CONCLUSIONS: The TIG is briefer, easier to apply and more efficient than the MEM. Possibly it is useful to detect cognitive impediment in clinical contexts. This study is preliminary. PMID- 11423043 TI - [Gustatory nervous pathway syndromes]. AB - Although the lingual nerve and the chorda tympani are the components of the classic peripheral gustatory pathway, loss of taste in patients after surgery for trigeminal neuralgia supports for the existence of an accessory gustatory pathway through the trigeminal sensory root and the gasserian ganglion. Bell's palsy is the most common pathology of the peripheral gustatory pathway. The central gustatory pathway ascends from the solitary tract nucleus in the medulla up to the upper pons in the ipsilateral central tegmental tract, rather than in the medial lemniscus as proposed in the past. It is not possible to specify whether the central gustatory pathway decussates or not at the lower midbrain level. Interruption of the gustatory pathway in the brainstem usually occurs with stroke or demyelination. The thalamic gustatory relay is located in the most medial aspect of the ventroposteromedial nucleus, immediately adjacent to the somatosensory area for the oral cavity and fingers. Therefore, ageusia associated with the sensory cheiro-oral syndrome may occur with a thalamic lesion. The laterality of the gustatory representation in the thalamus remains unresolved. Studies on epileptic gustatory aura have demonstrated that the insula and the anteromedial temporal lobe are the primary and secondary gustatory cortex, respectively. Taste perception results in patients with corpus callosum section and strokes or tumors involving the insula support the hypothesis that there is a gustatory representation of both hemitongues in the left cerebral hemisphere, whereas only the right hemitongue is represented in the right hemisphere. PMID- 11423044 TI - [On the duality of the sign of fanning of the toes]. AB - Starting from Babinski's original description, the author carries out a revision of semeiological value of the sign "fanning of the toes". It is considered that there are two clinical variants. The first, present in most cases of connatal encephalopathy, is due to involvement of the premotor area, which is usually accompanied by pyramidal, dystonic or athetoid signs. The second variant is present in cases of multiple sclerosis presumably attributable to involvement of the lateral spinal tracts including the reticulospinal one. The author proposes the need of using modern neuroimaging and electrophysiological techniques for further pathophysiological investigation of both clinical variants of the sign. PMID- 11423045 TI - [Brain abscess with air fluid level]. PMID- 11423046 TI - [Cerebral infarction due to spontaneous dissection of the left common carotid artery]. AB - Spontaneous dissection of the extracranial cervicocephalic arteries occurs most often in the internal carotid artery or vertebral artery. Spontaneous dissection of a common carotid artery is rare, with only nine cases having been reported. A 43-year-old man was hospitalized for sudden onset of motor aphasia and right arm weakness due to spontaneous dissection of a left common carotid artery; no sign of aortic disection was evident on aortogram. The most common causes of dissection of the common carotid artery are the extension of an aortic dissection and the complication in an angiogram with direct puncture; spontaneous dissection being very rare. We review the previous literature on this topic. PMID- 11423047 TI - [Intracranial hemorrhage from an arteriovenous malformation (AVM) in a tuberous sclerosis patient]. AB - We describe a previously diagnosed tuberous sclerosis patient that arrived to the hospital with an intracerebral hemorrhage whose origin was an arteriovenous malformation (AVM). A craniectomy was performed and the hematoma was evacuated with the alleviation of the patient. Although the association between tuberous sclerosis and intracerebral hemorrhage is known in the literature it has never been described associated with an AVM. PMID- 11423048 TI - [Main-in-the-barrel syndrome]. PMID- 11423049 TI - Intravascular ultrasound assessment of the stenoses location and morphology in the left main coronary artery in relation to anatomic left main length. AB - Eighty-seven left main stenoses were evaluated by angiography and intravascular ultrasound. Intravascular ultrasound analysis included left main length (bifurcation to ostium), stenosis location, stenosis length, stenosis external elastic membrane, lumen, plaque & media cross-sectional area (CSA), plaque burden (plaque & media/external elastic membrane CSA), calcium arc, calcium length, eccentricity, and remodeling index (stenosis/reference external elastic membrane CSA). Long anatomic left main arteries (length > or =10 mm, n = 43) were compared with short anatomic left main arteries (length <10 mm, n = 44) regarding stenosis location. Ostial (proximal third of left main artery) (n = 32) and nonostial (midthird and distal third) stenoses (n = 55) were compared regarding stenosis morphology. Short anatomic left main arteries developed stenoses more frequently near the ostium (ostium 55%, bifurcation 38%). Conversely, long anatomic left main arteries developed stenoses more frequently near the bifurcation (ostium 18%, bifurcation 77%, p = 0.001). Ostial left main stenoses were more common in women (44% vs 20%, p = 0.02), had larger lumen area (6.2 +/- 2.2 vs 4.6 +/- 2.3 mm(2), p = 0.002), less plaque burden (62 +/- 15% vs 80 +/- 9%, p <0.0001), less calcification (arc = 78 +/- 65 degrees vs 195 +/- 101 degrees, p <0.0001), and more negative remodeling (remodeling index = 0.87 +/- 0.19 vs 1.01 +/- 0.21, p = 0.005) than nonostial left main stenoses. Most ostial left main stenoses were categorized as eccentric (97% vs 76%, p = 0.01). Short and long left main arteries develop stenoses at different locations. Stenosis morphology was significantly different in these 2 locations. PMID- 11423050 TI - Simplified risk score models accurately predict the risk of major in-hospital complications following percutaneous coronary intervention. AB - The objectives of this analysis were to develop and validate simplified risk score models for predicting the risk of major in-hospital complications after percutaneous coronary intervention (PCI) in the era of widespread stenting and use of glycoprotein IIb/IIIa antagonists. We then sought to compare the performance of these simplified models with those of full logistic regression and neural network models. From January 1, 1997 to December 31, 1999, data were collected on 4,264 consecutive interventional procedures at a single center. Risk score models were derived from multiple logistic regression models using the first 2,804 cases and then validated on the final 1,460 cases. The area under the receiver operating characteristic (ROC) curve for the risk score model that predicted death was 0.86 compared with 0.85 for the multiple logistic model and 0.83 for the neural network model (validation set). For the combined end points of death, myocardial infarction, or bypass surgery, the corresponding areas under the ROC curves were 0.74, 0.78, and 0.81, respectively. Previously identified risk factors were confirmed in this analysis. The use of stents was associated with a decreased risk of in-hospital complications. Thus, risk score models can accurately predict the risk of major in-hospital complications after PCI. Their discriminatory power is comparable to those of logistic models and neural network models. Accurate bedside risk stratification may be achieved with these simple models. PMID- 11423051 TI - Long-term (> or =8 years) outcome after Palmaz-Schatz stent implantation. AB - The purpose of this single-center study was to evaluate the long-term (> or =8 years) outcome of Palmaz-Schatz intracoronary stenting and to identify independent predictors of outcome. Although short-term results of Palmaz-Schatz intracoronary stenting have been promising, with a reduction in both angiographic restenosis and clinical cardiac events up to 3 years, longer-term follow-up has not been established. We analyzed clinical outcome in 426 consecutive patients at least 8 years after coronary stenting. Demographic, clinical, and procedural predictors of restenosis, survival, and event-free survival, defined as freedom from death, myocardial infarction (MI), and coronary revascularization (target stented site, target vessel, and any revascularization) were analyzed. Before discharge, 28 patients (6.6%) sustained at least 1 major cardiovascular event: 3 deaths (0.7%), 18 MIs (4.2%), and 17 repeat revascularizations. Surviving patients were followed for 8.9 years (interquartile range 8.4 to 9.4). After discharge, 59 patients (13.9%) died, 47 (11.1%) sustained an MI, and 188 (44.4%) underwent coronary revascularization. The 8-year event-free survival (freedom from death, freedom from death/MI/target-stented site revascularization, and freedom from death/MI/any coronary revascularization) was (mean +/- SE) 0.86 +/- 0.01, 0.62 +/- 0.03, and 0.47 +/- 0.02, respectively. Unstable angina, lower left ventricular ejection fraction, and saphenous vein graft stenting were found to be independent predictors of death during follow-up. Hypertension, unstable angina, multivessel disease, and multiple stent implantation were found to be independent predictors of the composite of death/MI/any coronary revascularization during follow-up. This study provided a useful assessment of very long-term outcome in survival, event-free survival, and predictors of major cardiac events 8 to 10 years after Palmaz-Schatz stent implantation. PMID- 11423052 TI - Fractal correlation properties of heart rate dynamics and adverse events in patients with implantable cardioverter-defibrillators. AB - The aim of this study was to determine the prognostic significance of nonlinear and standard heart rate (HR) variability parameters in predicting future adverse events (AEs) in patients with implantable cardioverter-defibrillators. In postinfarction studies, nonlinear measures of HR variability obtained from long term electrocardiographic recordings have been suggested to be better predictors of adverse outcomes than conventional HR variability measures. Fifty-five high risk patients with reduced left ventricular function and an implantable cardioverter-defibrillator had a 10-minute, high-resolution electrocardiographic recording after which they were followed for 25 months on average. Implantable cardioverter-defibrillator shock or death was determined as the end point. The SD of all normal-to-normal RR intervals, the square root of the mean squared differences of successive normal-to-normal RR intervals, and the proportion of interval differences of successive normal-to-normal RR intervals >50 ms, low frequency and high-frequency powers of the power spectrum and their ratio were calculated as conventional measures of HR variability. The short-term scaling exponent (alpha(1)) and approximate entropy were determined as nonlinear measures of HR variability. AEs occurred in 23 patients (42%). Patients with AEs had significantly lower alpha(1) than event-free patients: 0.81 +/- 0.29 (mean +/- SD) versus 1.01 +/- 0.30 (p = 0.02). None of the other HR variability parameters differed significantly between patients with and without AEs. In the Cox proportional-hazards model including age, gender, ejection fraction, occurrence of ventricular tachyarrhythmia before defibrillator implantation, beta-blocker usage, and alpha(1), only alpha(1) was an independent predictor of AEs: hazard ratio 1.20 (95% confidence interval 1.03 to 1.39) for every 0.10 decrease in alpha(1) (p = 0.020). In conclusion, alpha(1) obtained from a 10-minute electrocardiographic recording yields important prognostic information about the risk of AEs in patients with implantable cardioverter-defibrillators. PMID- 11423053 TI - Effects of kaliuretic peptide on sodium and water excretion in persons with congestive heart failure. AB - Kaliuretic peptide, a 20-amino acid peptide hormone synthesized in the heart, enhances urine flow twofold, whereas atrial natriuretic peptide (ANP) enhances urine flow four- to 11-fold in healthy persons. The present investigation was designed to (1) determine whether kaliuretic peptide may have beneficial diuretic effects in persons with congestive heart failure (CHF), and (2) compare its beneficial effects with ANP in the treatment of CHF. Kaliuretic peptide (100 ng/kg body weight/min) given intravenously for 60 minutes to subjects with New York Heart Association class III CHF increased urine flow fourfold (p <0.001), which was maximal 212 hours after its infusion was stopped. Kaliuretic peptide enhanced sodium excretion threefold in subjects with CHF (p <0.01). Kaliuretic peptide increased the urinary excretion rate of potassium ion and fractional excretion of potassium 3.5- and twofold (p <0.05), respectively. ANP (same concentration) did not significantly enhance urine flow. ANP enhanced sodium excretion two- to sixfold in half of the CHF subjects, whereas it had no effect on sodium excretion in the other half. ANP did not significantly increase fractional excretion of sodium but did increase fractional excretion of potassium (p <0.05) during the first 20 minutes of its infusion. ANP-infused patients with CHF became hypotensive. None became hypotensive secondary to kaliuretic peptide. These data indicate that the diuretic properties of kaliuretic peptide in persons with CHF, as opposed to those of ANP, are not diminished (but rather are increased) compared with their effects in healthy persons. In patients with CHF, kaliuretic peptide causes a natriuresis-a feature not observed in those without sodium retention. PMID- 11423054 TI - Elevated plasma tissue plasminogen activator and anti-THP-1 antibodies are independently associated with decreased graft survival in cardiac transplant recipients. AB - Hemostatic and immunologic factors have been implicated in future cardiac events in patients with coronary artery disease. The role of these factors and their interaction is less established in cardiac transplant recipients. We sought to characterize the role of these factors in these patients. Cardiac transplant patients who presented for surveillance coronary angiography and/or endomyocardial biopsy were eligible for enrollment. Ninety-nine consecutive patients were enrolled. Plasma levels of tissue-type plasminogen activator (t PA), plasminogen activator inhibitor-1, von Willebrand factor, fibrin D-dimer, and anti-t-PA antibody were determined by enzyme-linked immunosorbent assays. Anti-THP-1 cell antibodies directed against a monocytic leukemia cell line were detected by incubating patient plasma with THP-1 cells. Bound antibody was detected using goat peroxidase-labeled immunoglobulin G directed against human immunoglobulins. Lipids were measured by enzymatic methods. Multivariate analysis identified the presence of anti-THP-1 cell antibodies (risk ratio 4.41, p = 0.002), t-PA antigen (risk ratio 1.10, p = 0.033), donor age 20 to 26 years (risk ratio 8.83, p = 0.042), and donor age >36 years (risk ratio 15.53, p = 0.009) as predictors of allograft failure. Altered hemostatic function, as demonstrated by elevated plasma t-PA antigen levels, is predictive of subsequent allograft failure in cardiac transplant recipients. In addition, the presence of anti-THP-1 cell antibodies in these patients is predictive of allograft failure. PMID- 11423055 TI - Comparison of the occurrence of ventricular arrhythmias in patients with acutely decompensated congestive heart failure receiving dobutamine versus nesiritide therapy. AB - Ventricular arrhythmias are common in patients with congestive heart failure (CHF) and may be exacerbated by positive inotropic therapy. Because human B-type natriuretic peptide (nesiritide), an arterial and venodilator, inhibits sympathetic activity, it may decrease the incidence of arrhythmias. Our investigation compares the arrhythmogenicity of dobutamine with nesiritide. A total of 305 patients with decompensated CHF requiring intravenous vasoactive therapy were randomized to receive standard therapy (n = 102) or nesiritide (0.015 microg/kg/min [n = 103] or 0.030 microg/kg/min [n = 100]) to gain additional data on the relative safety and efficacy of nesiritide compared with standard parenteral care. Dobutamine was chosen as the standard care agent in 58 subjects. During study drug infusion, all patients had continuous clinical hemodynamic and electrocardiographic monitoring. The dobutamine and nesiritide groups were similar with respect to baseline use of antiarrhythmic agents, including beta blockers. Serious arrhythmias and the incidence of cardiac arrest were more common in patients who received dobutamine than in those taking nesiritide: sustained ventricular tachycardia, 4 (7%) versus 2 (1%), respectively (p = 0.014); nonsustained ventricular tachycardia, 10 (17%) versus 23 (11%), respectively (p = 0.029); cardiac arrest, 3 (5%) versus 0, respectively (p = 0.011). We conclude that among patients with decompensated CHF for whom dobutamine is selected as standard therapy, the incidence of serious ventricular arrhythmias and cardiac arrest is significantly greater than the incidence of these events in patients randomized to nesiritide. PMID- 11423056 TI - Comparison of a daily fixed 2.5-mg warfarin dose with a 5-mg, international normalized ratio adjusted, warfarin dose initially following heart valve replacement. AB - Patients starting oral anticoagulant therapy after heart valve replacement initially require a lower target international normalized ratio (INR) (2.0, range 1.5 to 2.6) because of a higher risk of bleeding until pericardial wires are removed. In a previous retrospective analysis, we observed a higher sensitivity to warfarin in these patients compared with nonsurgical patients. In a randomized clinical trial, we compared a fixed, lower dose of warfarin (2.5 mg) with the standard treatment consisting of a 5-mg loading dose, then adjusted to the target INR during the first 5 days of anticoagulation. INRs were measured daily, but the fixed dose was only modified on day 3 if the INR was <1.5 or >3.0. One hundred ninety-seven patients were considered eligible for the study. The 2 groups were well matched according to age, gender, body mass index, concomitant treatments, and type of valves implanted. The proportion of INRs >2.6 during the study period was 42.5% in the 5-mg group and 26.2% in the 2.5-mg group (p <0.05), and the proportion of INRs >3.0 on day 3 was 23.9% and 9.5% (p <0.05), respectively. In the 2.5-mg group, 35.7% of patients had an INR <1.5 on day 3 and had the dose increased (vs 3.5%, p <0.001); however, in the 5-mg group, 95.6% had the initial dose reduced, 49.6% had the dose withheld for at least 1 day, and the mean dose during the 5 days of study was 3.08 mg. Average time to achieve therapeutic range was higher in the 2.5-mg group (2.72 vs 1.98 days, p <0.0001), but the approach to the targeted INR was more regular, and the gap between target and mean INR on day 5 was smaller. There were no bleeding or thromboembolic complications in either group. Thus, a lower loading dose of warfarin in patients after heart valve replacement reduces excessive anticoagulation and offers a more regular achievement of the therapeutic target by reducing the number of dose adjustments. Daily monitoring of the INR is still recommended. PMID- 11423057 TI - Comparison of valve resistance with effective orifice area regarding flow dependence. AB - Aortic valve resistance has been proposed to represent the severity of aortic stenosis because some studies observed that it was less affected by change in flow than the valve-effective orifice area, but this issue remains controversial. The objective of this study was to systematically analyze the theoretical and practical determinants of these parameters in relation to changes in flow. Valve area and resistance in different valves were studied in vitro in a pulse duplicator system at different flow rates and in vivo in 90 subjects referred to either exercise or dobutamine infusion. Theoretical analysis and experimental results both demonstrated a unique relation between resistance (RES), valve effective orifice area (EOA), and flow rate (Q): RES = K x (Q/EOA(2)). Accordingly, in fixed stenoses or in mechanical valves, resistance increased markedly with flow rate both in vitro (+0.88 +/- 0.26%/% of flow increase) and in vivo (mechanical valves: +2.09 +/- 4.61, fixed stenotic valves: +0.59 +/- 0.32%/%), whereas valve area did not change significantly (<0.2%/%). In contrast, in valves with a flexible orifice (bioprostheses and some patients with aortic stenosis), resistance was less increased due to the increase in valve area. Thus, both from a theoretical and a practical standpoint, valve resistance is much more flow dependent than valve area, particularly in fixed stenoses. Situations in which resistance does not increase with flow rate are unpredictable and are found in flexible valves when there is a concomitant increase in valve area. PMID- 11423058 TI - Differentiation between pathologic and physiologic left ventricular hypertrophy by tissue Doppler assessment of long-axis function in patients with hypertrophic cardiomyopathy or systemic hypertension and in athletes. AB - To identify new echocardiographic indexes of long-axis function that might differentiate between pathologic and physiologic left ventricular (LV) hypertrophy, we compared 60 subjects with different types of LV hypertrophy (group I: 15 patients with hypertrophic cardiomyopathy, group II: 15 patients with systemic hypertension, and group III: 30 athletes) with 20 normal subjects (group IV). The peak velocities of mitral annular motion at 4 sites were measured from the apex by tissue Doppler echocardiography. There were no differences in mean age and global ejection fraction between groups. Groups I and II had lower long-axis systolic and early diastolic velocities than the athletes (p <0.01) for all 4 sites. The best differentiation of pathologic from physiologic hypertrophy was provided by a mean systolic annular velocity <9 cm/s (sensitivity 87%, specificity 97%). Heterogeneity of annular velocities discriminated between group I and group II. Thus, long-axis systolic and early diastolic velocities are decreased in patients with pathologic hypertrophy, but preserved in athletes. These simple new echocardiographic parameters can differentiate between pathologic and physiologic hypertrophy. PMID- 11423059 TI - Metabolic effects of high-protein, low-carbohydrate diets. PMID- 11423060 TI - Emergency room administration of eptifibatide before primary angioplasty for ST elevation acute myocardial infarction and its effect on baseline coronary flow and procedure outcomes. PMID- 11423061 TI - Excellent prognosis in patients with unstable angina pectoris classified as "low risk" at admission despite presence of severe coronary artery disease. PMID- 11423062 TI - Influence of continuation of beta blockers during dobutamine stress echocardiography for the assessment of myocardial viability in patients with severe ischemic left ventricular dysfunction. PMID- 11423063 TI - Frequency of gross hematuria shortly after percutaneous coronary intervention. PMID- 11423064 TI - Clinical outcomes and costs associated with a first episode of uncomplicated atrial fibrillation presenting to the emergency room. PMID- 11423065 TI - Atrial fibrillation and mortality among patients with acute coronary syndromes without ST-segment elevation: results from the PURSUIT trial. PMID- 11423066 TI - Efficacy and safety of midodrine hydrochloride in patients with refractory vasovagal syncope. PMID- 11423067 TI - Intraventricular conduction delay and functional mitral regurgitation. PMID- 11423068 TI - Late development of atrioventricular block after congenital heart surgery in down syndrome. PMID- 11423069 TI - Influence of pravastatin on lipoproteins, and on endothelial, platelet, and inflammatory markers in subjects with peripheral artery disease. PMID- 11423070 TI - Gradient and spin echo(cardiographic) magnetic resonance imaging: prevalence of descriptive errors in published articles. PMID- 11423071 TI - Bed tilt as an effort-independent maneuver to improve patent foramen ovale assessment by transthoracic contrast echocardiography. PMID- 11423072 TI - Spontaneous fatal rupture of a coronary arterial aneurysm into the right ventricle*. PMID- 11423073 TI - Neuronal properties in cultured ultimobranchial C cells of chick embryos: process outgrowth and expression of TuJ1 and enkephalin. AB - We have analyzed the neuronal properties in cultured ultimobranchial C cells isolated from embryonic chicks at different developmental stages (12--16 days of incubation) by immunohistochemistry and electron microscopy. The ultimobranchial glands mostly consist of C cell solids. In 13-day-old embryos, many C cells cultured for 7 days on laminin-coated slides extended long neurite-like processes, reaching 300 microm in length. Neuritic outgrowth of C cells was regulated developmentally and virtually unaffected by nerve growth factor (NGF). The cultured C cells expressed intense immunoreactivity for calcitonin and enkephalin. It was also confirmed by confocal laser-scanning microscopy that almost all C cells were intensely immunostained by both the calcitonin antiserum and the monoclonal antibody TuJ1, a neuron-specific marker. Scanning electron microscopy identified the outgrowth of long, branching neuritic processes emerging from C cell soma. The processes had numerous varicosities along their course and ended in growth cones. The C cells with processes were usually monopolar and less frequently bipolar or multipolar. Transmission electron microscopy revealed the presence of membrane-bounded secretory granules in the cultured C cells. The neuritic processes of C cells contained aggregations of microfilaments, intermediate filaments and microtubules arranged in parallel to the long axis. In addition, synaptic-like contacts showing desmosome-like membrane-thickenings and accumulations of small clear vesicles and dense-cored vesicles were formed between the endings of the processes and the surface of C cells. These results indicate that the C cells cultured from early chick embryos (12- and 13-day-old) maintain the neuronal characteristics for long periods in vitro. PMID- 11423074 TI - Combination of inflammatory cytokines increases nitrite and nitrate levels in the paraventricular nucleus of conscious rats. AB - Inflammatory cytokines stimulate glial cells in vitro to produce nitric oxide (NO) from inducible NO synthase (iNOS). Whether the stimulation with cytokines produces NO derived from iNOS has not hitherto been demonstrated in the vivo brain. Nitrite and nitrate (NOx(-)) levels in the rat paraventricular nucleus (PVN) were measured before and after intraparenchymal microinjection of cytokines with a microdialysis technique. The cytokines, tumor necrosis factor (TNF)-alpha (10 ng), interleukin (IL)-1 beta (2 ng), and interferon (IFN)-gamma (2 ng) were microinjected. None of the cytokines alone had any effect on the NOx(-) levels for 8 h. But a combination of TNF-alpha and IFN-gamma gradually increased NOx(-) levels beginning at 140 min after the microinjection, and NOx(-) levels reached 1.8 times the basal level at 380 min. A combination of TNF-alpha and IL-1 beta increased NOx(-) beginning at 340 min, reaching 1.7 times the basal level at 440 min, whereas a combination of IL-1 beta and IFN-gamma had no effect. Microinjection of a mixture of all three cytokines increased NOx(-) levels beginning at 120 min, reaching 3.3 times the basal level at 400 min. Aminoguanidine, which is a selective inhibitor of iNOS, reduced NOx(-) levels induced by the mixed cytokine treatment. Semi-quantitative RT-PCR for iNOS mRNA was done. The intensity of the iNOS mRNA band for the cytokine-treated PVN was stronger than that for the vehicle-treated PVN. These results suggest that the increased NOx(-) after the treatment with mixed cytokines were dependent on iNOS activity. This is the first report to indicate that only cytokines induce NOS in vivo in the brain. PMID- 11423075 TI - Evaluation of the effect of stress on the blood--brain barrier: critical role of the brain perfusion time. AB - The Gulf war syndrome has drawn increased attention in the issue of the effect of stress on the blood-brain barrier (BBB). We have applied various stressful modalities and tested BBB disruption as measured by the amount of Evans blue (EB) retained by brain parenchyma. We have evaluated the retention of this marker as a function of the perfusion time of the brain following stress. This was done to distinguish between the marker retained in the lumen of small blood vessels and the marker retained by the brain parenchyma. Mice were exposed to either short swim stress or restraint stress. In mice exposed to either swim or restraint stress that were perfused for 1 min, the amount of EB retained in the brain was significantly higher as compared to non-stressed controls. Fifteen min perfusion markedly reduced the EB brain content to levels found in the non-stressed animals. In rats exposed to neural or metabolic stressful stimuli and perfused for 15 min, the EB content was similar to non-stressed controls. Our results demonstrate that various stress modalities have no effect on the BBB permeability and insufficient wash of blood vessels by perfusion may cause misinterpretation of permeability studies. PMID- 11423076 TI - Agonistic behavior in groups of limbic epileptic male rats: pattern of brain damage and moderating effects from normal rats. AB - Different ratios of normal male rats and male rats in which limbic seizures had been induced by a single systemic injection of lithium and pilocarpine were housed in groups of six. The group ratios ranged along the continuum from all normal rats to all experimental rats. The average numbers of episodes of boxing, biting and mounting--thrusting per rat per hour per group were recorded by direct observation (red light) for 1 h during the midscotophase. Groups that contained less than two normal rats exhibited significantly elevated amounts of agonistic (boxing, biting) behavior but not mounting behavior. Multiple regression analyses showed that combinations of neuronal loss within only two to three areas accommodated at least 50% of the variance in the numbers of these behaviors. PMID- 11423077 TI - Degradation of c-Fos protein expressed by N-methyl-D-aspartic acid in nuclear fractions of murine hippocampus. AB - In both nuclear and cytosolic fractions of murine hippocampus, constitutive expression was seen with Fra-2 protein, but not with other Fos family members tested including c-Fos, Fos-B and Fra-1 proteins. Fos-B protein was only detected in nuclear fractions. The systemic administration of N-methyl-D-aspartic acid (NMDA) induced marked and transient expression of c-Fos protein, but not other family members, in both hippocampal fractions 2 h later. In vitro incubation at 30 degrees C led to more rapid degradation of inducible c-Fos protein than constitutive Fra-2 protein in nuclear fractions obtained 2 h after the administration of NMDA, without significantly affecting that of both member proteins in cytosolic fractions. The addition of phosphatase inhibitors significantly delayed the initial degradation rate of inducible c-Fos protein, with concomitant facilitation of that of constitutive Fra-2 protein, in nuclear fractions. The addition of protease inhibitors also delayed the initial degradation of constitutive Fra-2 protein, without markedly altering that of inducible c-Fos protein, in nuclear fractions. Immunoprecipitation analysis revealed that NMDA induced phosphorylation of c-Fos protein on tyrosine residues in nuclear fractions to a lesser extent than that on serine residues 2 h after administration. These results suggest that NMDA signals may be propagated to the nucleus to induce both expression and degradation of c-Fos protein through a molecular mechanism associated with phosphorylation on serine and/or tyrosine residues in murine hippocampus. PMID- 11423078 TI - Neuroprotective effects of pramipexole in young and aged MPTP-treated mice. AB - This study examined the effect of pramipexole (PPX), a selective dopamine (DA) D(3)/D(2) agonist, on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced damage to the nigrostriatal dopamine system in young (8-week-old) and aged (12 month-old) mice. Co-administration of PPX and MPTP to young or aged mice, followed by 2 or 14 days of additional PPX treatment, significantly attenuated MPTP-induced striatal DA loss. Pramipexole treatment also significantly attenuated the loss of tyrosine hydroxylase immunoreactive neurons (TH-IR) within the substantia nigra pars compacta (SNc) in both young and aged animals. Effects of PPX administration on dopaminergic cell survival were confirmed in Nissl stained sections and by quantitation of retrogradely labeled Fluorogold-positive SNc neurons. Protective effects of PPX on striatal DA levels and SNc DA neuron survival were similar in young and aged animals, although the magnitude of these effects was significantly less in aged animals. These findings support the early initiation of PPX therapy in Parkinson's disease patients. PMID- 11423079 TI - Effects of perinatal undernourishment on neuronal development of the facial motor nucleus in the rat. AB - The facial nucleus (FN) of the rat is composed of multipolar neurons generated between gestational days G12 and G15. This nucleus is involved in the mechanisms underlying muscle contraction during the sucking reflex. After birth, the neuronal substrate of this reflex is gradually organized to allow the performance of other functions such as gnawing, chewing, swallowing and drinking. Undernourishment is known to produce different degrees of delayed brain development, the greatest of which are similar to the characteristics of the premature syndrome. Neuronal morphological alterations are associated with sucking-reflex deficiencies, which interfere with feeding by the newborn. The current study shows that perinatal undernourishment leads to dendritic arbor hypoplasia and small alterations of soma size in Golgi--Cox impregnated FN neurons of rats. The data suggest that these morphological alterations of FN neurons, may be associated with shifts in the input and integration of signals, and may finally modify the elaboration of motoneuron discharges partly modulating bucolabial muscle contraction during sucking movements and facial expression. Additionally, neonatal nutritional rehabilitation modifies the effects on FN neuronal development, ameliorating the influence of early adverse nutritional conditions. PMID- 11423080 TI - Differentiation in the immunocytochemical features of intrinsic and cortically projecting neurons in the rat claustrum -- combined immunocytochemical and axonal transport study. AB - Retrograde axonal transport method of the fluorescent tracer FluoroGold (FG) was combined with immunocytochemistry to investigate the occurrence of nitric oxide synthase (NOS), somatostatin (SOM), neuropeptide Y (NPY) and vasoactive intestinal peptide (VIP) in both intrinsic and cortically projecting neurons of the rat claustrum. Only NOS was detected in both the scattered projecting neurons and internal neurons of the claustrum. Approximately 20% of NOS-immunoreactive neurons in the claustrum were also retrogradely labeled with FG after tracer injections into the frontal cortex. The other substances were exclusively confined to the population of interneurons, which mainly displayed an oval, round or fusiform shape and a medium size. Apart from the neuronal somata, the proximal parts of the dendritic arborization were clearly visible. The immunoreactive neurons were randomly distributed in the claustrum and their neuronal size and shape did not differ in the various parts of the studied structure. Co localization of NOS and SOM or NOS and NPY was reported. In conclusion, SOM, VIP and NPY do not appear to play a significant role in the claustro-cortical projection but are most probably involved in modulation and information transfer in the claustrum. The appearance of NOS in both cortically projecting and intrinsic neurons of the claustrum may be indicative of a fundamentally different role in the functioning of the claustro-cortical loop. PMID- 11423081 TI - Hypodynamia--hypokinesia induced variations in expression of fos protein in structures related to somatosensory system in the rat. AB - There have been many reports describing modifications of the sensory and motor cortex following various types of disuse. Hypodynamia--hypokinesia is characterized by the absence of weight-bearing and by a decrease in motor activity. We have shown a reorganization of the cortical cartography after hypodynamia--hypokinesia. In order to give an anatomical account for this cortical plasticity, we set out to determine whether cerebral and spinal structures exhibited variations of their neuronal activation. For this purpose, immunocytochemical detection of Fos protein was performed in the rat brain and spinal cord. Following stimulation of the sciatic nerve, Fos protein was detected in the primary and secondary somatosensory cortex in control rats and in rats submitted to an episode of 14 days of hypodynamia--hypokinesia. Results showed that the stimulation of the sciatic nerve induced an increase in the number of Fos-immunoreactive neurons in all these structures. Moreover, after hypodynamia- hypokinesia, the number of Fos-immunoreactive neurons was increased in the primary and secondary somatosensory cortex and in the spinal cord. These results provide evidence for a higher activation of cortical cells after hypodynamia- hypokinesia in comparison to controls. These data support the hypothesis that hypodynamia--hypokinesia contributes to the development of functional plasticity. PMID- 11423082 TI - The effect of bone morphogenetic protein-7 (BMP-7) on functional recovery, local cerebral glucose utilization and blood flow after transient focal cerebral ischemia in rats. AB - Bone morphogenetic protein-7 (BMP-7) has been shown to enhance dendritic growth and improve functional recovery after experimental stroke. In this study, we examined the effect of BMP-7 on functional recovery, local cerebral blood flow (LCBF) and local cerebral glucose utilization (LCMRglu) following transient middle cerebral artery occlusion. Sprague--Dawley rats (n=29) were anesthetized with halothane/nitrous oxide and received 2-h middle cerebral artery occlusion (MCAo) by poly-L-lysine-coated intraluminal suture. Rectal and cranial temperatures were regulated at 37.0--37.5 degrees C. BMP-7 or vehicle (volume, 25 microl) was administered intracisternally in a blinded fashion at 24 h after MCAo. Neurological status was evaluated during occlusion (60 min) and daily for 2 days after MCAo. In matched animal groups, LCMRglu was measured autoradiographically with [(14)C]2-deoxyglucose (2-DG) and LCBF with [(14)C]iodoantipyrine 48 h after MCAo. Four animals groups were studied: LCMRglu series (BMP-7, n=7; vehicle, n=8); LCBF series (BMP-7, n=6; vehicle, n=8). Average three-dimensional image data sets were constructed for each group and were compared by pixel-based statistical methods. Rectal and cranial temperatures, mean blood pressure, plasma glucose and blood gases were similar among groups. BMP-7 significantly improved the total neurological score compared to vehicle at 48 h after MCAo (7.3+/-0.4 vs. 9.0+/-0.2, respectively; P<0.0003). Compared to vehicle-rats, BMP-7 enhanced glucose utilization in the basal ganglia ipsilateral to stroke and improved LCBF in ipsilateral subthalamus, but decreased LCBF and LCMRglu in contralateral cortical regions. PMID- 11423083 TI - Neuroprotective effect of aminoguanidine on transient focal ischaemia in the rat brain. AB - Using serial magnetic resonance imaging we have evaluated the effectiveness of aminoguanidine (AG) as a neuroprotective agent in a rat model of transient middle cerebral artery occlusion (MCAO). Because aminoguanidine's neuroprotective properties have primarily been ascribed to its action as iNOS inhibitor, we also performed a biochemical analysis of nitric oxide metabolites and NOS isoforms in our model of ischaemia. Daily injections of AG (100 mg/kg) or saline, were started at 6 h after the occlusion and the effects of this treatment on lesion progression monitored by T(2)-weighted MRI at 6 (pre-treatment scan), 24 and 72 h. Measurements of lesion volumes showed that between 6 and 72 h post-MCAO, lesion growth was slower in AG-treated rats than in control rats. This difference was most pronounced between 24 and 72 h post-MCAO when AG halted the lesion volume expansion observed in control rats. Measurements of plasma NOx (nitrite plus nitrate) at 0, 24, 48 and 72 h after MCAO, showed that NO levels did not differ significantly between the AG- and saline-treated groups at any time-point. Moreover, NOS activity assays revealed that no iNOS activity was present in any of the brains tested and that constitutive neuronal NOS activity was similar across the two hemispheres between both groups. The absence of iNOS protein in the ischaemic and contralateral hemispheres at 48 and 72 h after MCAO (control group only) was confirmed by Western blot analysis. These results suggest that AG treatment reduces the rate of growth of ischaemic lesions, perhaps preserving the functioning of perifocal neurons. Our observations contradict suggestions that high levels of NO generated by iNOS are partially responsible for exacerbating the neuronal damage in the postischaemic phase of MCAO. Although this does not rule out a role for AG as a neuroprotective agent via its ability to inhibit iNOS, these findings indicate that neuroprotective actions of AG may also be mediated via other cellular targets. PMID- 11423084 TI - A-134974: a novel adenosine kinase inhibitor, relieves tactile allodynia via spinal sites of action in peripheral nerve injured rats. AB - Extracellular levels of adenosine (ADO) can be raised through inhibition of adenosine kinase (AK), a primary metabolic enzyme for ADO. AK inhibitors have shown antinociceptive activity in a variety of animal models of nociception. The present study investigated the antinociceptive actions of a novel and selective AK inhibitor, A-134974 (IC(50)=60 pM), in a rat model of neuropathic pain (ligations of the L5/L6 spinal nerves) and explored the relative contributions of supraspinal, spinal and peripheral sites to the actions of A-134974. Systemic A 134974 dose-dependently reduced tactile allodynia (ED(50)=5 micromol/kg, i.p.) for up to 2 h. Fall latencies in the rotorod test of motor coordination were unaffected by systemic administration of A-134974 (at doses up to 30 micromol/kg, i.p.). Administration of A-134974 intrathecally (i.t.) was more potent (ED(50)=10 nmol) in relieving tactile allodynia than delivering the compound by intracerebroventricular (ED(50)>100 nmol, i.c.v.) or intraplantar (ED(50)>500 nmol) routes suggesting that spinal sites of action are the primary contributors to the anti-allodynic action of A-134974. The anti-allodynic effects of systemic A-134974 (10 micromol/kg, i.p.) were antagonized by the non-selective ADO receptor antagonist, theophylline (30-500 nmol) administered i.t. These data demonstrate that the novel AK inhibitor A-134974 potently reduces tactile allodynia through interactions with spinal sites and adds to the growing evidence that AK inhibitors may be useful as analgesic agents in a broad spectrum of pain states. PMID- 11423085 TI - Changes in striatal dopamine D3 receptor regulation during expression of and recovery from MPTP-induced parkinsonism. AB - Striatal dopamine (DA) D3 receptor density (measured by quantitative receptor autoradiography) and mRNA expression (measured by reverse transcriptase polymerase chain reaction) were analyzed in cats symptomatic for and recovered from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced parkinsonism. In symptomatic cats, D3 receptor density was significantly decreased in all regions of the caudate nucleus (CD) (66--77%), the nucleus accumbens (NACC) (52--83%) and the islands of calleja (IC) (67%), all of which returned to normal values in recovered cats. In contrast, D3 receptor mRNA expression was slightly elevated in symptomatic cats, and significantly increased above normal in recovered cats (45% increase in the CD and 91% in the NACC). Thus, reduction of parkinsonian signs was related to normalization of striatal D3 receptor number. These alterations in D3 receptor expression may play an important role in the recovery process observed in this model of parkinsonism. PMID- 11423086 TI - Unique responses of limbic met-enkephalin systems to low and high doses of methamphetamine. AB - A single administration of a low (0.5 mg/kg) or high (10 mg/kg) dose of methamphetamine (METH) significantly altered the met-enkephalin (M-Enk) systems associated with some, but not all, limbic structures examined. Neither treatment influenced M-Enk levels 3 h after drug exposure in any limbic region studied; however, 12 h after drug administration, 0.5 mg/kg of METH reduced the tissue content of this peptide in both the nucleus accumbens shell (NAs) and the frontal cortex (FrCx). This was similar to the effect of this treatment on the anterior striatal region. In contrast, the high dose of METH increased M-Enk content in the frontal cortex and anterior striatum (AS), but had no effect in the nucleus accumbens shell. By 24 h, the effects of METH in the anterior striatum subsided, but decreases in M-Enk levels were still observed after both the low- and the high-dose METH treatments in the nucleus accumbens shell. The levels of M-Enk were not changed at any of the time points examined in the core of the nucleus accumbens (NAc). In general, treatment with a low or high dose of METH causes distinct and regional selective changes in the tissue levels of M-Enk in the limbic system. These changes appear to be mediated by dopamine (DA) D(2) and D(1) receptor activation. PMID- 11423087 TI - The putative OP(4) antagonist, [Nphe(1)]nociceptin(1-13)NH(2), prevents the effects of nociceptin in neuropathic rats. AB - Nociceptin or orphanin FQ (N/OFQ) is the natural ligand of the opioid receptor like 1 receptor (ORL-1), which has been also classified as the fourth member of the opioid family of receptors and named OP(4). Elucidation of the biological role of N/OFQ has been hampered by the lack of compounds that selectively block the OP(4) receptor. Recently, a N/OFQ derivative, [Nphe(1)]N/OFQ(1-13)NH(2), has been found to possess OP(4) antagonistic properties both in vitro and in vivo models. We investigated its spinal effect in the chronic constriction injury of the sciatic nerve in the rat, a model relevant to neuropathic pain in humans. Intrathecal (i.t.) administration of N/OFQ (0.2--20 nmoles) dose-dependently reversed mechanical allodynic-like behavior, while [Nphe(1)]N/OFQ(1-13)NH(2) (20- 120 nmoles, i.t.) was ineffective on its own. [Nphe(1)]N/OFQ(1-13)NH(2) (60--120 nmoles, i.t.) antagonized N/OFQ (about 80% of reduction) but did not modify the activity of morphine (20 nmoles, i.t.). These results further support, for the first time in a chronic model of pain, the specific antagonistic profile of [Nphe(1)]N/OFQ(1-13)NH(2)vs the OP(4) receptor. This pseudopeptide is an interesting pharmacological tool to better clarify the role of N/OFQ in pathophysiology. PMID- 11423088 TI - MAP2 phosphorylation and visual plasticity in Xenopus. AB - Microtubule-associated protein 2 (MAP2) has been implicated in activity-dependent structural changes in dendrites. MAP2 regulates the assembly of cytoskeletal proteins such as microtubules and actin, and its function is phosphorylation dependent. In hippocampus, MAP2 has been reported to be dephosphorylated by activation of the NMDA-type glutamate receptor, a key player in synaptic plasticity. In this work, we used a phospho-specific MAP2 antibody (Ab 305) that recognizes epitopes close to the microtubule-binding domain to investigate the possible role of MAP2 in the Xenopus visual system. The binocular system in Xenopus exhibits activity-dependent synapse rearrangement during a critical period of development. We have found that, in critical period animals, NMDA receptor activation leads to the dephosphorylation of MAP2 at sites recognized by Ab 305 in a dose-dependent manner. We compared the responses of MAP2 to NMDA treatment in animals with high binocular plasticity (critical period juveniles and dark-reared adults) and low plasticity (normal adults). Our results show that, in all groups, NMDA treatment induces the dephosphorylation of MAP2. Tecta from frogs with different degrees of plasticity show no differences in the baseline level of MAP2 phosphorylation or in the NMDA-induced MAP2 dephosphorylation response. These results suggest that activity may modify dendrite structure via the NMDA receptor--MAP2-cytoskeletal protein pathway, but this pathway does not seem to be a determinant of the degree of plasticity. PMID- 11423089 TI - Locomotor behavior of dopamine D1 receptor transgenic/D2 receptor deficient hybrid mice. AB - Mice that incorporate the dopamine D1 receptor transgene controlled by the D1 receptor promoter exhibit a marked increase of D1 binding in several extra striatal brain regions and show a paradoxical hypokinetic response to D1 agonist [Exp. Neurol. 157 (1999) 169]. The agonist-induced locomotor behavior of D1 receptor transgenic mice is similar to baseline locomotor activity manifested by D2 receptor deficient mice [J. Neurosci. 18 (1998) 3470]. The similarity between these two behavioral phenotypes raised the possibility that stimulation of the over-expressed D1 receptors in the transgenic mice could cause a suppression of D2 receptor responses that manifest in hypokinesia. Alternatively, the similar phenotypes could result from altered D1/D2 receptor balance in both animal models. Two different approaches were undertaken to test these alternative hypotheses. (1) The effects of pharmacological blockade of D2 receptors on D1 agonist-stimulated hypokinesia of the D1 over-expressing animals were investigated. (2) The behavioral phenotype of hybrid D1 receptor over expressing/D2 receptor deficient mice generated by crossbreeding the D2 knockout mice and the D1 transgenic animals was studied. The results of these studies suggested that the hypomotor response of the D1 transgenic mice was not a result of an interaction of the over-expressed D1 receptors with the native D2 receptors and that over-expressed D1 receptors likely mediate hypokinesia in the D1 transgenic animals. Considering the significance of the D1 dopamine receptor as a therapeutic target for Parkinson's disease, this D1 receptor over-expressing model provides an important experimental system to probe the basis for altered behavioral responses following stimulation of transgenetically up-regulated receptors. PMID- 11423090 TI - Macrophage invasion into injured cochlear nerve and its modification by methylprednisolone. AB - Post-traumatic invasion of macrophages into the cochlear nerve of the rat and measurement of how their invasion was modified by the administration of methylprednisolone were investigated for the first time by using a reproducible and quantifiable experimental model of cochlear nerve injury. Two weeks after precise cochlear nerve compression, a massive invasion of ED1 immunostained macrophages was observed at the compressed portion of the cochlear nerve, and this invasion of macrophages was markedly reduced in the rats to which methylprednisolone had been administered during the pre- and post-compression period. Concomitantly, the residual number of spiral ganglion cells was found to be greater in the compression+methylprednisolone group than in the control compression group. The tissue loss observed in the lesion epicenter was also significantly less in the compression+methylprednisolone group than in the control compression group. The results of our present study demonstrated the effectiveness of methylprednisolone treatment to ameliorate trauma induced cochlear nerve degeneration in the acute phase. However, these results may reflect the sum effects of methylprednisolone on macrophages, including both its beneficial effect by inhibiting the negative aspects of macrophages through attenuating macrophage recruitment to the lesion site, and at the same time an undesirable effect by sacrificing the positive aspects of macrophage function. Moreover, one reservation should be added that the protective effects of steroid to injured cochlear nerve may have operated via a pathway not related to macrophage function. Besides macrophages, various cells and factors participate in the process of CNS injury, and their effects may potentially work either positively or negatively with respect to CNS protection and regeneration at each particular time during the on-going process of CNS injury. Therefore, future investigation in CNS injury should be directed toward understanding such complex mechanisms involved in this process. PMID- 11423091 TI - Regulation of expression and enzymatic activities of tyrosine and tryptophan hydroxylases in rat brain after acute electroconvulsive shock. AB - Acute electroconvulsive shock (ECS) causes a significant increase of protein synthesis in depressive patients and such an increase raises the possibility that the regulation of specific proteins and enzymatic activities in the brain might be one of the mechanisms required for the induction of long-term adaptive neurochemical changes after electroconvulsive therapy. In current studies, we investigated and compared simultaneously the short- and long-term effects of an acute ECS on the expression and enzymatic activities of both tyrosine and tryptophan hydroxylases (TH and TpOH, respectively) in different rat brain areas. Our results demonstrated that an acute ECS produced: (1) a long-lasting decrease in TH and TpOH protein levels in locus ceruleus (LC), ventral tegmental area (VTA) and in TpOH protein level in the raphe centralis (RC), maximal at 72 h, with concomitant changes in mRNA levels and enzymatic activities in the LC only; (2) large increase of TpOH protein levels in the frontal cortex (Cxf) (+145%) and increase of TH protein levels in the hippocampus (Hip) (+207%), maximal at 72 h and 7 days which was not accompanied by corresponding increase of in vivo enzymatic activities. Furthermore, a second ECS increased in vivo TpOH activity in the Cxf (+19%) while decreasing K(m) value (-50%) for tetrahydrobiopterin cofactor. A stability of the observed findings on TpOH activity in the Cxf after repeated ECS might be one of the mechanisms for the antidepressant effects of electroconvulsive therapy. PMID- 11423092 TI - Learning-related interpositus activity is conserved across species as studied during eyeblink conditioning in the rat. AB - Single-unit activity was monitored in the interpositus nucleus of the cerebellum during standard delay conditioning of the eyeblink response in freely-moving rats. The rats were implanted with recording electrodes in the interpositus nucleus then received paired presentations of a tone-conditioned stimulus (CS) and eye-shock unconditioned stimulus during acquisition training. The acquisition training was followed by CS-alone extinction training. Learning-related activity in the interpositus nucleus developed over the course of acquisition training and then activity returned to baseline levels during subsequent extinction training. These findings are consistent with rabbit studies that have demonstrated similar changes in neuronal activity in the interpositus nucleus over the course of acquisition and extinction of the eyeblink response, thus providing strong evidence for the generality of the neural substrates of eyeblink conditioning across species. PMID- 11423093 TI - Involvement of the cerebellar adenosine A(1) receptor in cannabinoid-induced motor incoordination in the acute and tolerant state in mice. AB - Cannabinoids are known to impair motor function in humans and laboratory animals. We have demonstrated an accentuation of cannabinoid (CP55,940)-induced motor incoordination in mice by the adenosine A(1) receptor-selective agonist N(6) cyclohexyladenosine (CHA) (4 ng) using an intracerebellar (ICB) microinjection method. This effect was mediated by the A(1) receptor because pre-treatment with ICB 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) (100 ng), an adenosine A(1) receptor selective antagonist, completely abolished the accentuation. Furthermore, ICB pre-treatment with DPCPX (100 ng) before ICB CP55,940 (15 microg) attenuated the motor incoordination suggesting a modulation by an endogenous adenosine A(1) system. ICB microinjection of CHA or DPCPX prior to ICB vehicle had no effect on normal motor coordination. ICB microinjection of dipyridamole (25 microg), an adenosine transport inhibitor, significantly accentuated the motor incoordination by ICB CP55,940 (15 microg), providing further support for the involvement of endogenous adenosine in the action of CP55,940. Tolerance to the motor incoordinating effect of ICB CP55,940 was demonstrated following 3 days of i.p. CP55,940 (0.1, 1 or 2 mg/kg every 12 or 24 h; total of six or three injections, respectively). Interestingly, animals which exhibited tolerance to ICB CP55,940 also demonstrated tolerance to the accentuating effect of ICB CHA suggesting cross-tolerance between adenosine agonists and cannabinoids. Cross-tolerance was also demonstrated following 3 days of i.p. CHA (0.25 or 1 mg/kg every 24 h; total of three injections) as further evidence of the modulatory role of the cerebellar adenosine system in the acute manifestation of CP55,940-induced motor incoordination. The involvement of cerebellar adenosine and the A(1) receptor in cannabinoid actions is circumstantially supported by previous evidence that CB(1) receptors and A(1) receptors are both localized on cerebellar granule cell parallel fiber terminals and basket cell neurons where they serve to inhibit the release of neurotransmitters. PMID- 11423094 TI - N-Methyl-D-aspartate-induced c-Fos expression is enhanced in the forebrain structures related to emotion in Fyn-deficient mice. AB - Fyn tyrosine kinase is thought to be involved in the control of neuronal intracellular signal transduction elicited by neurotransmitter stimulation. Emotional disorders, such as fearfulness in Fyn-deficient mice, prompted us to investigate the neural mechanisms that lead to defective emotional expression by using functional neuroanatomical methods. In order to examine the reactivity of a specific neural network to excitatory neurotransmitter administration, we mapped the distribution of c-Fos-immunoreactive neurons after administering N-methyl-D aspartate (NMDA) to control and mutant mice at the subthreshold dose for seizure induction. The induction of neuronal c-Fos-immunoreactivity by NMDA was enhanced in the Fyn-deficient mice, and there was a much greater increase in immunopositive neurons in certain well-defined areas, such as the amygdaloid medial nuclear subdivisions, hypothalamic paraventricular nucleus, and midbrain periaqueductal gray, of the mutant. NMDA-induced c-Fos expression was attenuated by pretreatment with D-(-)-2-amino-5-phosphonovaleric acid, a competitive NMDA antagonist, both in the control and the mutant mice. These findings suggest that the excitability of the projection system from the amygdala to the hypothalamus and midbrain, the main pathways of emotional expression, is enhanced in Fyn deficient mice. PMID- 11423095 TI - Toxic effects of MPP(+) and MPTP in PC12 cells independent of reactive oxygen species formation. AB - MPTP is a toxin presumed to damage dopamine-secreting neurons by an oxygen free radical-mediated mechanism. Two steps in MPTP metabolism are the primary candidates for oxygen free radical generation: (a) MPTP oxidation to MPP(+) by a monoamine oxidase and (b) NADH dehydrogenase inhibition by MPP(+). In order to test the idea that MPTP toxicity is mediated by oxygen free radicals, we assessed lipid peroxidation and the effects of antioxidants in dopaminergic PC12 cells treated with MPTP or MPP(+). For comparison purposes, we also examined the effects of the pro-oxidant tert-butyl-hydroperoxide (TBHP) and of the dopaminergic toxin 6-hydroxydopamine (6-OHDA) in PC12 cells. MPTP and MPP(+), unlike TBHP, failed to induce lipid peroxidation in PC12 cells after a 4-h exposure. All toxins tested (MPTP, MPP(+), TBHP and 6-OHDA) caused a dose dependent decrease in [(3)H]dopamine ((3)H-DA) uptake in PC12 cultures. The hydroperoxide scavengers glutathione and N-acetyl-cysteine and the superoxide and peroxide scavenger EUK-134 protected PC12 cells from TBHP- and 6-OHDA-induced decrease in (3)H-DA uptake. However, no protection by these antioxidants at various concentrations and time regimens was observed against MPTP- or MPP(+) induced decreases in (3)H-DA uptake in PC12 cells. In addition, incubation of PC12 cells with the energy-rich substrate, NADH, attenuated MPP(+)-induced decrease in (3)H-DA uptake. These results suggest that MPTP-induced toxicity in dopaminergic PC12 cell cultures, does not involve oxygen free radical production, but rather may be caused by impairment in energy metabolism. PMID- 11423096 TI - Prism adaptation of reaching is dependent on the type of visual feedback of hand and target position. AB - The present study demonstrated that the magnitude of after-effect due to wedge prisms depends on the form of the visual feedback used to represent hand and target position in fast, targeted, transverse reaches. Trained human subjects made reaches with and without prisms in three visuomotor representations (VR): (1) the subject's actual hand and targets (Direct), (2) a real-time video broadcast of hand and targets (Video), or (3) abstract, computer-generated targets and a cursor representing hand position (Cursor). A significant after effect occurred in each VR. However, the magnitude of the after-effect was significantly different among VRs: the magnitude was greatest in Direct, smaller in Video and smallest in Cursor. A significant after-effect (carryover) also occurred when a subject prism-adapted reaches in one VR and then removed the prisms and made initial reaches in another VR. Our data showed that when reaches were prism-adapted in Direct and then prisms were removed, there was a large carryover to initial reaches in Video or Cursor (D-->V and D-->C). In contrast, when prisms were worn in Video and removed for reaches in Direct (V-->D), there was a significantly smaller carryover than from both D-->V and D-->C. Finally, when prisms were worn in Cursor and removed for reaches in Direct (C-->D), there was very little detectable carryover. Our results suggest that adaptation is context-dependent and that the magnitude of carryover is dependent on the VR in which adaptation occurred. Interpretations of adaptations made in abstract training and experimental conditions may be greatly affected by this finding. PMID- 11423097 TI - Reduction of cerebrospinal fluid amyloid beta after systemic administration of M1 muscarinic agonists. AB - Overproduction of the peptide amyloid beta (A beta) is a critical event in Alzheimer's disease (AD). Systemic administration of 3 M1-selective muscarinic agonists, AF102B, AF150S and AF267B, decreased cerebrospinal fluid (CSF) A beta concentrations; levels of CSF secreted beta-APP were not significantly altered. Rabbits treated for 5 days with s.c. injections of each drug (2 mg/kg/day) had levels of CSF A beta which were between 55 and 71% of control for A beta 1-40 and between 59 and 84% of control for A beta 1--42. PMID- 11423098 TI - Entorhinal cortex projection cells to the hippocampal formation in vitro. AB - Retrogradely labeled cells in superficial and deep layers of the entorhinal cortex (EC) were analyzed following application of the fluorescent tracer rhodamine-dextran-amine in different sites of the hippocampal formation in a slice preparation. The results demonstrate a strong projection from layer IV/V to the dentate gyrus, that is in slices significantly stronger than that from layer II. In deep layers a large number of multipolar cells were found which were only labeled by dye application to the subiculum. Patch-clamp recordings from these cells revealed intrinsic low threshold membrane potential oscillations, suggesting their possible contribution to oscillatory network activity of the EC and subiculum. PMID- 11423099 TI - Immunocytochemical detection of superoxide dismutases (SODs) in the periodontal Ruffini endings of the rat incisor. AB - The expression of immunoreactivities for superoxide dismutases (SODs), Mn-SOD and Cu/Zn-SOD, was immunohistochemically investigated in the lingual periodontal ligament and toe pads of adult rats. Immunocytochemistry for SODs revealed that the axon terminals of both the periodontal Ruffini endings and cutaneous Meissner's corpuscles showed mitochondrial Mn-SOD immunoreactivity, but not cytosolic Cu/Zn-SOD immunoreactivity, indicating Mn-SOD is a useful marker for identifying the mechanoreceptors. It is likely that Mn-SOD in the axon terminals of mechanoreceptors exerts protective action against nerve injury and neuronal death under severe conditions, serving to scavenge free radicals from the axon terminals. PMID- 11423100 TI - Kappa opioid receptor density is consistent along the rostrocaudal axis of the female rat spinal cord. AB - Kappa opioid receptors (KORs) were immunocytochemically localized at four different levels of the spinal cord of normally-cycling female rats in estrus or diestrus. KOR labeling was primarily observed in fine processes and a few neuronal cell bodies in the superficial dorsal horn and the dorsolateral funiculus. Quantitative light microscopic densitometry of the superficial dorsal horn revealed that there were no significant differences in KOR densities among spinal segments C1--C2, T2, T13--L1, and L6--S1 in either the estrus or diestrus phases. These results suggest that the potential for KOR-mediated antinociceptive responses is consistent along the rostrocaudal axis of the female rat spinal cord. PMID- 11423101 TI - The expression and cellular localization of phospholipase D1 in the rodent retina. AB - Phospholipase D (PLD) is one of the intracellular signal transduction enzymes and plays an important role in a variety of cellular functions. We investigated the expression and cellular localization of the PLD isozyme PLD1 in the rodent retina. Western blot analysis showed the presence of PLD1 at the protein level in the rat, mouse and guinea pig retinas. PLD1 immunoreactivity was localized in all Muller cells. Thus, PLD1 protein appears to be important in the functions of these cells in the rodent retina. PMID- 11423102 TI - Post-cultured development of basic electrophysiological properties of spinal neurons obtained from rat embryo. AB - Basic electrical profiles of cultured neurons are modified by multiple factors, such as cell growth, differentiation and cell damage from the isolation procedure. In the present study, we assessed development of electrophysiological properties of rat spinal neurons over the late embryonic and early postnatal period in a neuron-enriched culture. After recovery from acute damage within 2 days after plating, the input conductance and amplitudes of voltage-gated Na(+) and K(+) currents increased parallel to the increase in the cell capacitance. Whether this depended on the period or the growth of the cell area was estimated by normalizing the parameters with the cell capacitance. The input conductance per unit area, the membrane time constant and the Na(+) current density remained constant for two weeks. However, densities of two types of outward K(+) currents, an A-current and a delayed rectifier, required 3--5 days to reach the maximum, although neither thresholds for activation nor sensitivities to blockers (TEA and 4AP) altered. The hyperpolarizing shift of the resting membrane potential became stabilized within 6--8 days, suggesting that the development of the K(+) currents underlay the shift. These results show that passive electrical properties and voltage-gated currents of rat spinal neurons in the neuron-rich culture differ in temporal patterns of development but stabilize at the latest within a week, corresponding to the day of birth. PMID- 11423103 TI - Up-regulation of E2F-1 in Down's syndrome brain exhibiting neuropathological features of Alzheimer-type dementia. AB - We studied the expression of the apoptosis-related protein, E2F-1, in Down's syndrome (DS) brains. The immunoreactivity for E2F-1 was detected in the pyramidal neurons of the cerebral cortex from DS brains exhibiting the neuropathological features of dementia of Alzheimer type (DAT), in accordance with the amyloid beta protein (A beta) deposition in the neuron. Therefore, the implication is that A beta deposition may trigger E2F-1-mediated neuronal apoptosis in DS brains with DAT. PMID- 11423104 TI - Changes in the expression of glial glutamate transporters in the rat brain accompanied with morphine dependence and naloxone-precipitated withdrawal. AB - The expression of mRNAs for the glial glutamate transporters, GLT-1 and GLAST, in the rat brain accompanied with morphine dependence and naloxone-precipitated withdrawal was investigated by Northern blot analysis. The expression of GLT-1 mRNA was significantly decreased in the striatum and thalamus of morphine dependent rats, and significantly increased in the striatum 2 h after the naloxone-precipitated withdrawal, compared with that of naive rats. On the other hand, there were no significant changes in GLAST mRNA level in any brain region. These results suggest the involvement of GLT-1 in the development of morphine dependence and the expression of morphine withdrawal. PMID- 11423105 TI - Synthesis and antibacterial activities of quaternary ammonium salt of chitosan. AB - Chitosan derivatives with quaternary ammonium salt, such as N,N,N-trimethyl chitosan, N-N-propyl-N,N-dimethyl chitosan and N-furfuryl-N,N-dimethyl chitosan were prepared using different 96% deacetylated chitosan of M(v) 2.14x10(5), 1.9x10(4), 7.8x10(3). Amino groups on chitosan react with aldehydes to from a Schiff base intermediate. Quaternized chitosan were obtained by reaction of a Schiff base with methyl iodide. The yields, degree of quaternization and water solubility of quaternized chitosan were influenced by the molecular weight of the chitosan sample. The antibacterial activities of quaternized chitosan against Escherichia coli were explored by calculation of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) in water, 0.25 and 0.50% acetic acid medium. Results show the antibacterial activities of quaternized chitosan against E. coli is related to its molecular weight. Antibacterial activities of quaternized chitosan in acetic acid medium is stronger than that in water. Their antibacterial activities is increased as the concentration of acetic acid is increased. It was also found that the antibacterial activity of quaternized chitosan against E. coli is stronger than that of chitosan. PMID- 11423106 TI - The identification of the catalytic nucleophiles of two beta-galactosidases from glycoside hydrolase family 35. AB - The beta-galactosidases from Xanthomonas manihotis (beta-Gal Xmn) and Bacillus circulans (beta-Gal-3 Bcir) are retaining glycosidases that hydrolyze glycosidic bonds through a double displacement mechanism involving a covalent glycosyl enzyme intermediate. The mechanism-based inactivator 2,4-dinitrophenyl 2-deoxy-2 fluoro-beta-D-galactopyranoside was shown to inactivate beta-Gal Xmn and beta-Gal 3 Bcir through the accumulation of 2-deoxy-2-fluorogalactosyl enzyme intermediates with half lives of 40 and 625 h, respectively. Peptic digestion of these labeled enzymes and analysis by LC-MS identified Glu(260) and Glu(233) as the catalytic nucleophiles involved in the formation of the glycosyl-enzyme intermediate during catalysis by beta-Gal Xmn and beta-Gal-3 Bcir, respectively. These findings confirm the previous prediction of the position of these residues based on primary sequence similarities to other members of the glycoside hydrolase family 35. PMID- 11423107 TI - Dextran acceptor reaction of Streptococcus sobrinus glucosyltransferase GTF-I as revealed by using uniformly 13C-labeled sucrose. AB - A sucrose glucosyltransferase GTF-I from cariogenic Streptococcus sobrinus transferred the uniformly 13C-labeled glucosyl residue ([U-(13)C]Glc) from [U (13)C]sucrose to exogenous dextran T500 at the non-reducing-end, mostly by alpha (1-->6) linkages and partially by alpha-(1-->3) linkages, as revealed by the 13C (13)C NMR coupling pattern. With increasing amounts of [U-(13)C]sucrose, transfer of [U-(13)C]Glc to the alpha-(1-->3)-linked chain became predominant without increase in the number of chains. The transfer of [U-(13)C]Glc to an isomaltopentaose acceptor occurred similarly to its transfer to T500. alpha-(1- >3)-branches in the [U-(13)C]dextran, specifically synthesized from [U (13)C]sucrose by a Streptococcus bovis dextransucrase, were not formed by GTF-I, as judged by the observation that a newly-formed alpha-1,3,6-branched [U (13)C]Glc was not detected, which could have been formed by transferring the unlabeled Glc from sucrose to the internal alpha-(1-->6)-linked [U-(13)C]Glc at C 3. The 13C-(13)C one-bond coupling constants (1J) were also recorded for the C-1- C-6 bond of the internal alpha-(1-->6)-linked [U-(13)C]Glc and of the non reducing-end [U-(13)C]Glc. PMID- 11423108 TI - Purification and characterization of a beta-glucuronidase from Aspergillus niger. AB - A beta-glucuronidase from Pectinex Ultra SP-L, a commercial pectolytic enzyme preparation from Aspergillus niger, was purified 170-fold by ion-exchange chromatography and gel filtration. Apparent M(r) of the purified enzyme, estimated by denaturing gel electrophoresis and size-exclusion chromatography, were 68,000 and 71,000, respectively, indicating that the enzyme is a monomeric protein. It released uronic acids not only from p-nitrophenyl beta-glucosiduronic acid (PNP-GlcA) but also from acidic galactooligosaccharides carrying either beta D-glucosyluronic or 4-O-methyl-beta-D-glucosyluronic residues at the nonreducing termini through beta-(1-->6)-glycosidic linkages. The enzyme exhibited a maximal activity toward these substrates at pH 3.0. A regioisomer, 3-O-beta glucosyluronic acid-galactose, was unsusceptible to the enzyme. The enzyme did act on a polymer substrate, releasing uronic acid from the carbohydrate portion of a radish arabinogalactan-protein modified by treatment with fungal alpha-L arabinofuranosidase. The enzyme produced acidic oligosaccharides by transglycosylation, catalyzing the transfer of uronic acid residues of PNP-GlcA and 6-O-beta-glucosyluronic acid-galactose to certain exogenous acceptor sugars such as Gal, N-acetylgalactosamine, Glc, and xylose. PMID- 11423109 TI - Structure of a colitose-containing O-specific polysaccharide of the marine bacterium Pseudoalteromonas tetraodonis IAM 14160(T). AB - O-specific polysaccharide was isolated by mild acid degradation of the lipopolysaccharide of Pseudoalteromonas tetraodonis type strain IAM 14160(T) and studied by sugar and methylation analyses along with 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, 1H,(13)C HMQC and HMBC experiments. The polysaccharide was found to consist of hexasaccharide repeating units containing one residue each of D-Gal, D-GlcA, D-GalNAc and D-GlcNAc and two residues of 3,6-dideoxy-L xylo-hexose (colitose, Col) and having the following structure:In common with the polysaccharides of some other bacteria, the polysaccharide studied contains a tetrasaccharide fragment alpha-Colp-(1-->2)-beta-D-Galp-(1-->3)-[alpha-Colp-(1- >4)]-beta-D-GlcpNAc, which is a colitose ('3-deoxy-L-fucose') analogue of the Lewis(b) blood group antigenic determinant. PMID- 11423110 TI - Specific degradation of pectins via a carbodiimide-mediated Lossen rearrangement of methyl esterified galacturonic acid residues. AB - A specific, chemical degradation of the methyl esterified galacturonic acid residues of pectins is described. These residues are converted, with hydroxylamine, to hydroxamic acids, and then, with a carbodiimide, to isoureas; the latter undergo a Lossen rearrangement on alkaline hydrolysis. The isocyanates formed are hydrolysed to 5-aminoarabinopyranose derivatives, which spontaneously ring open to give 1,5-dialdehydes. The latter are reduced, in situ, to avoid peeling reactions, with sodium borohydride to give substituted arabitol residues. Thus, overall, partially esterified pectins are specifically cleaved to generate a series of oligogalacturonic acids bearing an arabitol residue as aglycone. Analysis of oligomers so generated discloses the pattern of contiguous nonesterification in a variety of pectins of differing degrees of esterification. Other potential applications are described. PMID- 11423111 TI - Mild tagging procedures for the structural analysis of glycans. AB - The reductive oxyamination of model glycan structures has been investigated as a mild, alternative tagging procedure to reductive amination using O-(4 nitrobenzyl)-hydroxylamine. Oxime formation was quantitative, but the reduction step did not always go to completion. Novel O- and N-substituted 7 hydroxycoumaryl- and 3-methoxybenzylhydroxylamines were synthesized and shown to couple quantitatively with model saccharides by oxime formation and reductive hydroxyamination, respectively, under very mild, aqueous conditions. The fluorescent derivatives produced show good chromatographic and mass spectrometric properties. Both procedures are suitable for the labeling of carbohydrates and oligosaccharide fragments from glycosaminoglycan structures, such as heparin and heparan sulfate. PMID- 11423112 TI - Biotin labeling of the symbiotically important succinoglycan oligosaccharides of Rhizobium meliloti for identification of putative plant receptors. AB - The symbiotically important trimer of the succinoglycan octasaccharide subunit was labeled with a biotin tag through coupling with a 6-biotinamidohexan hydrazide and subsequent reduction with borane. The acetyl and succinyl groups in the molecule were stable to the two-step sequence, while a small percentage of the ketal in the pyruvate groups was reduced to an ether-linked lactic acid moiety attached to either the O-4 or O-6 position of the sugar residue under the reaction conditions. PMID- 11423113 TI - Structural features of arabinogalactan-proteins from the fruit of Lycium chinense Mill. AB - In addition to the other arabinogalactan-proteins (AGPs) (Cp-1-C and -D) already reported, two kinds of AGP (Cp-2-B and Hp-2-C) were obtained from the fruit of Lycium chinense Mill. The ratio of arabinose to galactose was approximately 1:1 in both samples, and the carbohydrate was linked O-glycosidically to serine in Cp 2-B, and to both serine and threonine residues of the protein in Hp-2-C. The weight-average molecular weight was 71,000 for Cp-2-B and 120,000 for Hp-2-C. Both samples also contained non-reducing terminal 3-O- and 4-O-substituted galacturonic acids. The ratio of 6-O-substituted galactose (linear part) and 3,6 di-O-substituted galactose (branching point) was almost unity in both samples, being obviously different from the case of Cp-1-C (predominant in the branching domain) and Cp-1-D (predominant in the linear domain). These results offer fresh insight into the grouping of the AGPs, based on the ratio of 6-O- and 3,6-di-O substituted galactosyl residues. PMID- 11423114 TI - Structural analysis of a new glycosphingolipid from the lipopolysaccharide lacking bacterium Sphingomonas adhaesiva. AB - A new glycosphingolipid, GSL-4B, was isolated from Sphingomonas adhaesiva and found to share the ceramide moiety with GSL-1 and GSL-3 from Sphingomonas capsulata studied earlier [Kawahara, K.; Moll, H.; Knirel, Y. A.; Seydel, U.; Zahringer, U. Eur. J. Biochem. 2000, 267, 1837-1846]. It is heterogeneous with respect to the long-chain bases erythro-2-amino-1,3-octadecanediol (sphinganine), (13Z)-erythro-2-amino-13-eicosene-1,3-diol, and (13Z)-erythro-2-amino-13,14 methylene-1,3-eicosanediol which in GSL-4B are present in the ratios of 1.1:1.0:1.1, and all bearing amide-linked (S)-2-hydroxymyristic acid. Methylation analysis and MALDI-TOF-MS along with 1H and 13C NMR spectroscopy showed that the carbohydrate part of GSL-4B has the structure of alpha-D-Glcp-(1-->4)-alpha-D Galp-(1-->6)-alpha-D-Glcp-(1-->4)-alpha-D-GlcpA-(1-->1)-Cer PMID- 11423116 TI - Genes modulated by histone acetylation as new effectors of butyrate activity. AB - A wealth of evidence correlates the chemopreventive activity of a fiber-rich diet with the production of butyrate. In order to identify the genes transcriptionally modulated by the molecule, we analyzed the expression profile of butyrate-treated colon cancer cells by means of cDNA expression arrays. Moreover, the effect of trichostatin A, a specific histone deacetylase inhibitor, was studied. A superimposable group of 23 genes out of 588 investigated is modulated by both butyrate and trichostatin A. Among them, a major target was tob-1, a gene involved in the control of cell cycle. tob-1 is also up-regulated by butyrate in a neuroblastoma-derived cell line, and its overexpression in the colon cells caused growth arrest. Our findings represent an extensive analysis of genes modulated by butyrate and identify completely new effectors of its biological activities. PMID- 11423117 TI - Different effects of the Ca(2+)-binding protein, KChIP1, on two Kv4 subfamily members, Kv4.1 and Kv4.2. AB - The Ca(2+)-binding protein, K(+) channel-interacting protein 1 (KChIP1), modulates Kv4 channels. We show here that KChIP1 affects Kv4.1 and Kv4.2 currents differently. KChIP1 slows Kv4.2 inactivation but accelerates the Kv4.1 inactivation time course. Kv4.2 activation is shifted in a hyperpolarizing direction, whereas a depolarizing shift occurs for Kv4.1. On the other hand, KChIP1 increases the current amplitudes and accelerates recovery from inactivation of both currents. An involvement of the Kv4 N-terminus in these differential effects is demonstrated using chimeras of Kv4.2 and Kv4.1. These results reveal a novel interaction of KChIP1 with these two Kv4 members. This represents a mechanism to further increase the functional diversity of K(+) channels. PMID- 11423118 TI - A synthetic peptide corresponding to the 550-585 region of alpha-dystroglycan binds beta-dystroglycan as revealed by NMR spectroscopy. AB - We have probed the binding of a synthetic peptide corresponding to the region 550 585 of the alpha subunit of dystroglycan with a recombinant protein fragment corresponding to the N-terminal extracellular region of beta-dystroglycan (654 750), using NMR in solution. In a 30:1 molar ratio, the peptide binds to the recombinant protein fragment in the fast/intermediate exchange regime. By monitoring the peptide intra-residue HN-Halpha peak volumes of the 2D TOCSY NMR spectra, both in the absence and in the presence of the recombinant fragment, we determined the differential binding affinities of each amino acid. We found that the residues in the region 550-565 (SWVQFNSNSQLMYGLP) are more influenced by the presence of the protein, whereas the C-terminal portion is marginally involved. These NMR results have been confirmed by solid-phase binding assays. PMID- 11423119 TI - Requirements of basic amino acid residues within the lectin-like domain of LOX-1 for the binding of oxidized low-density lipoprotein. AB - Lectin-like OxLDL receptor-1 (LOX-1) was identified as the major receptor for oxidized low-density lipoprotein (OxLDL) in aortic endothelial cells. LOX-1 is a type II membrane protein that structurally belongs to the C-type lectin family. Here, we found that the lectin-like domain of LOX-1 is essential for ligand binding, but the neck domain is not. In particular, the large loop between the third and fourth cysteine of the lectin-like domain plays a critical role for OxLDL binding as well as C-terminal end residues. Alanine-directed mutagenesis of the basic amino acid residues around this region revealed that all of the basic residues are involved in OxLDL binding. Simultaneous mutations of these basic residues almost abolished the OxLDL-binding activity of LOX-1. Electrostatic interaction between basic residues in the lectin-like domain of LOX-1 and negatively charged OxLDL is critical for the binding activity of LOX-1. PMID- 11423120 TI - Green fluorescent protein-based halide indicators with improved chloride and iodide affinities. AB - The green fluorescent protein YFP-H148Q is sensitive to halides by a mechanism involving halide binding and a shift in pK(a). However, a limitation of YFP-H148Q is its low halide sensitivity, with K(d)>100 mM for Cl(-). Indicators with improved sensitivities are needed for cell transport studies, particularly in drug discovery by high-throughput screening, and for measurement of Cl(-) concentration in subcellular organelles. YFP-H148Q libraries were generated in which pairs of residues in the vicinity of the halide binding site were randomly mutated. An automated procedure was developed to screen bacterial colonies for improved halide sensitivity. Analysis of 1536 clones revealed improved anion sensitivities with K(d) down to 2 mM for I(-) (I152L), 40 mM for Cl(-) (V163S), and 10 mM for NO(3)(-) (I152L). The anion-sensitive mechanism of these indicators was established and their utility in cells was demonstrated using transfected cells expressing the cystic fibrosis transmembrane conductance regulator chloride channel. PMID- 11423121 TI - Activation of p38MAPK by TGF-beta in fetal rat hepatocytes requires radical oxygen production, but is dispensable for cell death. AB - We have previously found that transforming growth factor-beta (TGF-beta) induces an increase in radical oxygen species (ROS) production that mediates its apoptotic effects in fetal hepatocytes. In this paper we show that TGF-beta activates p38 mitogen-activated protein kinase (p38MAPK) and ROS may be responsible for this activation. Activation of p38MAPK occurs late, coincident with the maximal production of ROS, it is inhibited by radical scavengers and it is accentuated by the presence of glutathione synthesis inhibitors. However, p38MAPK does not appear to be involved in any of the apoptotic events: loss of Bcl-x(L) levels, cytochrome c release, cleavage of caspase substrates and loss of cell viability. PMID- 11423122 TI - Mathematical analysis of a mechanism for autonomous metabolic oscillations in continuous culture of Saccharomyces cerevisiae. AB - Autonomous metabolic oscillations were observed in aerobic continuous culture of Saccharomyces cerevisiae. Experimental investigation of the underlying mechanism revealed that several pathways and regulatory couplings are involved. Here a hypothetical mechanism including the sulfate assimilation pathway, ethanol degradation and respiration is transformed into a mathematical model. Simulations confirm the ability of the model to produce limit cycle oscillations which reproduce most of the characteristic features of the system. PMID- 11423123 TI - A novel plant alpha4-fucosyltransferase (Vaccinium myrtillus L.) synthesises the Lewis(a) adhesion determinant. AB - We have partially characterised an alpha4-fucosyltransferase (alpha4-FucT) from Vaccinium myrtillus, which catalysed the biosynthesis of the Lewis(a) adhesion determinant. The enzyme was stable up to 50 degrees C. The optimum pH was 7.0, both in the presence and in the absence of Mn(2+). The enzyme was inhibited by Mn(2+) and Co(2+), and showed resistance towards inhibition with N ethylmaleimide. It transferred fucose to N-acetylglucosamine in the type I Galbeta3GlcNAc motif from oligosaccharides linked to a hydrophobic tail and glycoproteins (containing the type I motif). Sialylated oligosaccharides containing the type II Galbeta4GlcNAc motif were not acceptors. The catalytic mechanism of the plant alpha4-FucT possibly involves a His residue, and it must have arisen by convergent evolution relative to its mammalian counterparts. PMID- 11423124 TI - HSF1/hsp72 pathway as an endogenous anti-inflammatory system. AB - We investigated the occurrence and the role of HSF1 activation and inducible hsp72 expression in the carrageenin pleurisy in the rat. Molecular analysis performed on pleural cells collected from rat pleural cavity after carrageenin challenge revealed increased HSF1 activation and hsp72 expression. Moreover, local injection of a double-stranded oligodeoxynucleotide (ODN) containing the heat shock element sequence, acting as transcription factor decoy, exacerbated the inflammatory reaction. The exacerbation, induced by wild-type, but not by mutant ODN decoy, was associated to both inhibition of HSF1/DNA binding activity and reduction of hsp72 expression. In conclusion, this study shows that HSF1 activation and hsp72 expression both actually occur in acute inflammation and that the remission of the inflammatory reaction is tightly associated to the HSF1 dependent hsp72 expression, suggesting a a relevant role for the HSF1/hsp72 pathway as an endogenous anti-inflammatory system. PMID- 11423125 TI - Transglutaminase-mediated cross-linking of alpha-crystallin: structural and functional consequences. AB - Aggregation and covalent cross-linking of the crystallins, the major structural proteins of the eye lens, increase light scattering by the lens leading to opacification and cataract. Disturbance of calcium homeostasis in the tissue is one of the factors implicated in cataractogenesis. Calcium-activated transglutaminase (TG)-catalyzed cross-linking of some lens proteins has been reported earlier. We show here that alpha-crystallin, a major structural protein in the lens and a member of the small heat shock protein family, is also a substrate for TG-mediated cross-linking, indicating the presence of donor Lys and acceptor Gln residues in the protein. Upon TG-catalyzed dimerization, the secondary and tertiary structures of the protein are altered, and its surface hydrophobicity reduced. The chaperone-like property of the protein, suspected to be one of its functions in situ, is substantially reduced upon such cross linking. These results, taken together with earlier ones on lens beta-crystallins and vimentin, suggest that TG-mediated events might compromise lens function. Also, since alpha-crystallin occurs not only in the lens but in other tissues as well, such TG-catalyzed cross-linking and the associated alterations in its structure and activity would be of general pathological interest. PMID- 11423126 TI - The fission yeast meiotic regulator Mei2p undergoes nucleocytoplasmic shuttling. AB - Schizosaccharomyces pombe Mei2p is an RNA-binding protein that switches the cell cycle from mitotic to meiotic. Mei2p forms a unique dot in the nucleus prior to meiosis I, aided by a non-coding RNA molecule termed meiRNA. Here we show that Mei2p intrinsically undergoes nucleocytoplasmic shuttling. Artificial acceleration of nuclear migration of Mei2p advances nuclear dot formation, but meiRNA does not appear to promote the dot formation by modulating the migration rate of Mei2p into the nucleus. Rather, this RNA is likely to facilitate the assembly of Mei2p into a dot structure and trap the protein as such in the nucleus. PMID- 11423127 TI - Novel peptides from assassin bugs (Hemiptera: Reduviidae): isolation, chemical and biological characterization. AB - Three novel peptides were isolated from the venomous saliva of predatory reduviids. They were identified by mass spectrometry and HPLC analysis and consist of 34-36 amino acid residues. They are relatively homologous to the calcium channel blockers omega-conotoxins from marine cone snails and belong to the four-loop Cys scaffold structural class. Ptu1, the shortest peptide, was chemically synthesized (sPtu1) and co-eluted with its native form. Circular dichroism spectra of the sPtu1 showed a high content of beta-turns similar to that of omega-conotoxins GVIA and MVIIA. Electrophysiological experiments demonstrated that sPtu1 reversibly blocks the N-type calcium channels expressed in BHK cells. PMID- 11423128 TI - Fibronectin is a binding partner for the myelin-associated glycoprotein (siglec 4a). AB - The myelin-associated glycoprotein (MAG) mediates cell-cell interactions between myelinating glial cells and neurons. Here we describe the extracellular matrix glycoprotein fibronectin as a binding partner of MAG. It has been identified by affinity precipitation with MAG-Fc from NG108-15 cells and by microsequencing of two peptides derived from a 210-kDa protein band. Western blot analysis showed that fibronectin is also present in MAG binding partners isolated from N(2)A (murine neuroblastoma) cells, rat brain and rat spinal cord. Different fibronectin isoforms have been isolated from brains of young and adult rats, indicating that the expression of MAG binding fibronectin changes during development. PMID- 11423129 TI - Amyloid-like fibrils from an 18-residue peptide analogue of a part of the central domain of the B-family of silkmoth chorion proteins. AB - Chorion is the major component of silkmoth eggshell. More than 95% of its dry mass consists of the A and B families of low molecular weight structural proteins, which have remarkable mechanical and chemical properties, and protect the oocyte and the developing embryo from the environment. We present data from negative staining, Congo red binding, X-ray diffraction, Fourier transform-Raman, attenuated total reflectance infrared spectroscopy and modelling studies of a synthetic peptide analogue of a part of the central domain of the B family of silkmoth chorion proteins, indicating that this peptide folds and self-assembles, forming amyloid-like fibrils. These results support further our proposal, based on experimental data from a synthetic peptide analogue of the central domain of the A family of chorion proteins, that silkmoth chorion is a natural, protective amyloid [Iconomidou et al., FEBS Lett. 479 (2000) 141-145]. PMID- 11423130 TI - Homeodomain proteins Mox1 and Mox2 associate with Pax1 and Pax3 transcription factors. AB - Mox1 and Mox2 homeobox genes have been shown to be critical in axial skeleton and in limb muscle development respectively. Pax1 and Pax3 gene products are also implicated in these processes. Mox and Pax expression patterns are highly overlapping both spatially and temporally during embryonic development. We show here for the first time that Mox proteins physically interact with Pax1 and Pax3 using the yeast two-hybrid protein interaction assay as well as in vitro biochemical assays. There is a strong preference of Mox1 to associate with Pax1 rather than Pax3 and of Mox2 to associate with Pax3 rather than Pax1. The observed interactions are mediated through the homeodomain of Mox. PMID- 11423132 TI - The Holy Grail of wood evolution - from wood anatomy to tissue-specific gene expression: to what extent do molecular studies of biosynthesis of cell wall biopolymers help the understanding of the evolution of woody species? PMID- 11423133 TI - Approaches to understanding the functional architecture of the plant cell wall. AB - Cell wall polysaccharides are some of the most complex biopolymers known, and yet their functions remain largely mysterious. Advances in imaging methods permit direct visualisation of the molecular architecture of cell walls and the modifications that occur to polymers during growth and development. To address the structural and functional relationships of individual cell wall components, we need to better characterise a broad range of structural and architectural alterations in cell walls, appearing as a consequence of developmental regulation, environmental adaptation or genetic modification. We have developed a rapid method to screen large numbers of plants for a broad range of cell wall phenotypes using Fourier transform infrared microspectroscopy and Principal Component Analysis. We are using model systems to uncover the genes that encode some of the cell-wall-related biosynthetic and hydrolytic enzymes, and structural proteins. PMID- 11423134 TI - Sucrose synthase localizes to cellulose synthesis sites in tracheary elements. AB - The synthesis of crystalline cellulose microfibrils in plants is a highly coordinated process that occurs at the interface of the cortex, plasma membrane, and cell wall. There is evidence that cellulose biogenesis is facilitated by the interaction of several proteins, but the details are just beginning to be understood. In particular, sucrose synthase, microtubules, and actin have been proposed to possibly associate with cellulose synthases (microfibril terminal complexes) in the plasma membrane. Differentiating tracheary elements of Zinnia elegans L. were used as a model system to determine the localization of sucrose synthase and actin in relation to the plasma membrane and its underlying microtubules during the deposition of patterned, cellulose-rich secondary walls. Cortical actin occurs with similar density both between and under secondary wall thickenings. In contrast, sucrose synthase is highly enriched near the plasma membrane and the microtubules under the secondary wall thickenings. Both actin and sucrose synthase lie closer to the plasma membrane than the microtubules. These results show that the preferential localization of sucrose synthase at sites of high-rate cellulose synthesis can be generalized beyond cotton fibers, and they establish a spatial context for further work on a multi-protein complex that may facilitate secondary wall cellulose synthesis. PMID- 11423135 TI - Cell-wall architecture and lignin composition of wheat developed in a microgravity environment. AB - The microgravity environment encountered during space-flight has long been considered to affect plant growth and developmental processes, including cell wall biopolymer composition and content. As a prelude to studying how microgravity is perceived - and acted upon - by plants, it was first instructive to investigate what gross effects on plant growth and development occurred in microgravity. Thus, wheat seedlings were exposed to microgravity on board the space shuttle Discovery (STS-51) for a 10 day duration, and these specimens were compared with their counterparts grown on Earth under the same conditions (e.g. controls). First, the primary roots of the wheat that developed under both microgravity and 1 g on Earth were examined to assess the role of gravity on cellulose microfibril (CMF) organization and secondary wall thickening patterns. Using a quick freeze/deep etch technique, this revealed that the cell wall CMFs of the space-grown wheat maintained the same organization as their 1 g-grown counterparts. That is, in all instances, CMFs were randomly interwoven with each other in the outermost layers (farthest removed from the plasma membrane), and parallel to each other within the individual strata immediately adjacent to the plasma membranes. The CMF angle in the innermost stratum relative to the immediately adjacent stratum was ca 80 degrees in both the space and Earth-grown plants. Second, all plants grown in microgravity had roots that grew downwards into the agar; they did not display "wandering" and upward growth as previously reported by others. Third, the space-grown wheat also developed normal protoxylem and metaxylem vessel elements with secondary thickening patterns ranging from spiral to regular pit to reticulate thickenings. Fourthly, both the space- and Earth-grown plants were essentially of the same size and height, and their lignin analyses revealed no substantial differences in their amounts and composition regardless of the gravitational field experienced, i.e. for the purposes of this study, all plants were essentially identical. These results suggest that the microgravity environment itself at best only slightly affected either cell wall biopolymer synthesis or the deposition of CMFs, in contrast to previous assertions. PMID- 11423136 TI - Induced compression wood formation in Douglas fir (Pseudotsuga menziesii) in microgravity. AB - In the microgravity environment of the Space Shuttle Columbia (Life and Microgravity Mission STS-78), were grown 1-year-old Douglas fir and loblolly pine plants in a NASA plant growth facility. Several plants were harnessed (at 45 degrees ) to establish if compression wood biosynthesis, involving altered cellulose and lignin deposition and cell wall structure would occur under those conditions of induced mechanical stress. Selected plants were harnessed at day 2 in orbit, with stem sections of specific plants harvested and fixed for subsequent microscopic analyses on days 8, 10 and 15. At the end of the total space mission period (17 days), the remaining healthy harnessed plants and their vertical (upright) controls were harvested and fixed on earth. All harnessed (at 45 degrees ) plant specimens, whether grown at 1 g or in microgravity, formed compression wood. Moreover, not only the cambial cells but also the developing tracheid cells underwent significant morphological changes. This indicated that the developing tracheids from the primary cell wall expansion stage to the fully lignified maturation stage are involved in the perception and transduction of the stimuli stipulating the need for alteration of cell wall architecture. It is thus apparent that, even in a microgravity environment, woody plants can make appropriate corrections to compensate for stress gradients introduced by mechanical bending, thereby enabling compression wood to be formed. The evolutionary implications of these findings are discussed in terms of "variability" in cell wall biosynthesis. PMID- 11423137 TI - Lignification and lignin topochemistry - an ultrastructural view. AB - This review discuses the ultrastructural aspects of cell wall lignification and lignin topochemistry. Lignification results from the enzyme mediated polymerization of monolignols initiated by unknown factors (initiation sites) located at the corners of cells and in the middle lamella. Lignification results in the filling of pores within the carbohydrate matrix following a sequence from the outer regions of the wall towards the lumen. The amount and chemical characteristics of lignin vary across the cell wall, with the presence of reaction wood, and among cell types. PMID- 11423138 TI - Reassessment of qualitative changes in lignification of transgenic tobacco plants and their impact on cell wall assembly. AB - In tobacco plants the effect of antisense down-regulation of various genes encoding enzymes of the monolignol biosynthetic pathway resulted in quantitative and qualitative changes in lignin distribution and in diverse alterations of the secondary wall assembly of modified tobacco plants. Total lignin content, composition in syringyl and guaiacyl units, and absolute proportions of condensed and non-condensed substructures occurring in the cell walls, were differentially modified according to the repressed gene. Immunocytochemical characterisation and visualisation of the distribution of condensed and non-condensed lignin substructure epitopes in transmission electron microscopy (TEM) revealed that some transformations entailed profound and specific alterations in the secondary wall biogenesis. Correlation between micro-morphological cell wall alterations and semi-quantitative immuno-analysis of the topochemical distribution of lignin sub-units suggests that the mode of polymerisation of monolignols into non condensed units, favoured by the microfibril matrix of the secondary wall, plays an important part in the lignified cell wall assembly. PMID- 11423139 TI - Dirigent proteins and dirigent sites in lignifying tissues. AB - Tissue-specific dirigent protein gene expression and associated dirigent (site) localization were examined in various organs of Forsythia intermedia using tissue printing, in situ mRNA hybridization and immunolabeling techniques, respectively. Dirigent protein gene expression was primarily noted in the undifferentiated cambial regions of stem sections, whereas dirigent protein sites were detected mainly in the vascular cambium and ray parenchyma cell initials. Immunolocalization also revealed cross-reactivity with particular regions of the lignified cell walls, these being coincident with the known sites of initiation of lignin deposition. These latter regions are considered to harbor contiguous arrays of dirigent (monomer binding) sites for initiation of lignin biopolymer assembly. Dirigent protein mRNA expression was also localized in the vascular regions of roots and petioles, whereas in leaves the dirigent sites were primarily associated with the palisade layers and the vascular bundle. That is, dirigent protein mediated lignan biosynthesis was initiated primarily in the cambium and ray cell initial regions of stems as well as in the leaf palisade layers, this being in accordance with the occurrence of the lignans for defense purposes. Within lignified secondary xylem cell walls, however, dirigent sites were primarily localized in the S(1) sublayer and compound middle lamella, these being coincident with previously established sites for initiation of macromolecular lignin biosynthesis. Once initiation occurs, lignification is proposed to continue through template polymerization. PMID- 11423140 TI - In situ hybridization and immunolocalization of lignan reductases in woody tissues: implications for heartwood formation and other forms of vascular tissue preservation. AB - Vascular plants have evolved with remarkable ways to form and protect the vasculature apparatus. In certain woody shrubs, the secondary xylem can have within its center a hollowed pith surrounded by secondary xylem, whereas in most trees there is a solid core of heartwood. Both types of woody systems have, however, the commonality of accumulating so-called 'secondary' metabolites, albeit to different extents, whose roles are to protect and preserve the vascular (lignified) tissues. This investigation had as its purpose establishing the nature of the cells involved in the biosynthesis of these specialized 'secondary' metabolites in plants forming heartwood and hollow piths, respectively. This was achieved by determining the tissue-specific expression of two lignan biosynthetic pathway enzymes: pinoresinol-lariciresinol reductase (PLR) and phenylcoumaran benzylic ether reductase (PCBER), soluble enzymes which catalyze analogous benzylic ether reductions of 8-8' and 8-5' linked lignans, respectively. Using Forsythia intermedia, reverse transcription-PCR (RT-PCR) and northern blots revealed that PLR mRNA accumulated mainly in young stems, as well as in young roots and petioles. Furthermore, PLR-specific DIG-labeled riboprobes established that in the stems its mRNA accumulated in the radial parenchyma cells [and to a lesser extent in the developing vessels], as well as in the cambial cells of developing secondary xylem. In addition, immunocytochemical localization of PCBER in Pinus taeda established that it was in the axial and radial parenchyma cells of secondary xylem of stems. That is, irrespective of whether the woody plants formed hollowed piths or heartwood, the 'secondary' metabolite pathways leading to the protective lignans predominantly involved axial and radial parenchyma cells. This is in contrast to monolignol coupling (i.e. the entry point to both the lignans and lignins), which appears to be more restricted to the vascular cambial regions. PMID- 11423141 TI - Biosynthesis and subcellular distribution of hydrolyzable tannins. AB - Pathways to complex gallotannins have been elucidated by enzyme studies, indicating that beta-glucogallin is required as principal acyl donor. Evidence for the in vitro oxidation of pentagalloylglucose, the pivotal metabolite in this sequence, to ellagitannins, is presented. Immunohistochemical studies with antibodies raised against pentagalloylglucose and the galloyltransferase catalyzing the formation of this ester revealed that leaf mesophyll cell walls were a typical site of origin and deposition of hydrolyzable tannins. Seasonal changes of these compounds were studied with extracts from cell walls and intracellular space of oak leaves. PMID- 11423142 TI - Pectins: structure, biosynthesis, and oligogalacturonide-related signaling. AB - Pectin is a family of complex polysaccharides present in all plant primary cell walls. The complicated structure of the pectic polysaccharides, and the retention by plants of the large number of genes required to synthesize pectin, suggests that pectins have multiple functions in plant growth and development. In this review we summarize the current level of understanding of pectin primary and tertiary structure, and describe new methods that may be useful to study localized pectin structure in the plant cell wall. We also discuss progress in our understanding of how pectin is biosynthesized and review the biological activities and possible modes of action of pectic oligosaccharides referred to as oligogalacturonides. We present our view of critical questions regarding pectin structure, biosynthesis, and function that need to be addressed in the coming decade. As the plant community works towards understanding the functions of the tens of thousands of genes expressed by plants, a large number of those genes are likely to be involved in the synthesis, turnover, biological activity, and restructuring of pectin. A combination of genetic, molecular, biochemical and chemical approaches will be necessary to fully understand the function and biosynthesis of pectin. PMID- 11423143 TI - Pectic polysaccharide rhamnogalacturonan II is covalently linked to homogalacturonan. AB - A borate-containing pectin was solubilized from sugar beet (Beta vulgaris L. ) cell walls by treatment with 0.5 M imidazole, pH 7. The molecular weight of the pectin was reduced when the borate ester was hydrolyzed by treatment with 1 N HCl. Treatment of the acid-treated pectin with boric acid in the presence of Pb(2+) gave a product whose molecular weight distribution was similar to the imidazole-soluble pectin. The imidazole-soluble pectin was saponified and then digested with endo- and exo-polygalacturonases. These treatments shifted the boron peak at the high molecular weight region to the low molecular weight (10 kDa), which corresponds to rhamnogalacturonan II-borate ester cross-linked dimer (dRG-II-B). The treatment also generated rhamnogalacturonan I (RG-I), dRG-II-B, monomeric rhamnogalacturonan II and galacturonic acid. These results show that imidazole solubilizes a high molecular weight borate-containing pectic complex composed of homogalacturonan-rhamnogalacturonan II and RG-I. Our data suggest that borate esters formed between rhamnogalacturonan II molecules cross-link the macromolecular pectin. PMID- 11423144 TI - Cell wall biosynthesis: glycan containing oligomers in developing cotton fibers, cotton fabric, wood and paper. AB - A series of oligomeric glycans can be extracted from the cell walls of developing cotton fibers with weak acid. Glycans that produce similar profiles on high pH anion chromatography with pulsed amperometric detection (HPAEC-PAD) are also found in a protein complex extracted from developing fibers and in amorphous aggregates found in association with immature fibers in developing, but not in mature cotton bolls. The quantity and composition of the glycans recovered from the carbohydrate-protein complex varies significantly with the time of day when the bolls are harvested. This diurnal variation is consistent with the hypothesis that secondary cell walls are deposited primarily at night. Incubation of re hydrated cotton fibers in the presence of exogenous oligosaccharides, myo inositol and glycerol substantially alters the apparent quantity of the oligomers extracted from the fibers. The same and similar glycans have also been extracted from cotton fabric, marine algae, various paper products and wood. While many of the oligomers isolated from the various cellulose sources display the same peaks by HPAEC-PAD, the specific number of oligomers and their relative quantities appear unique for each source of cellulosic material. Oligomeric glycans, as described in the preceding, are present in all cellulose sources that have been investigated. Their relative abundance changes in response to source, stage of development and other physiological variables. We hypothesize that the glycans are intermediates in the biological assembly of cellulose, and that their incorporation in cellulose is mediated by physicochemical and enzymatic mechanisms. PMID- 11423145 TI - Bonding of hydroxycinnamic acids to lignin: ferulic and p-coumaric acids are predominantly linked at the benzyl position of lignin, not the beta-position, in grass cell walls. AB - A suspension in dichloromethane-water (18:1, v/v) of various fractions containing hydroxycinnamic acid ester-ether bridges between lignin and polysaccharides prepared from cell walls of matured oat (Avena sativa L.) intemodes, and a solution of their acetates in the same solvent, were treated with 2,3-dichloro 5,6-dicyano-1,4-benzoquinone (DDQ). This reagent selectively cleaves benzyl ether and ester linkages of negatively charged aromatic nuclei. The sample treated with DDQ was directly hydrolysed either under mild (1 M NaOH, overnight at 37 degrees C) or severe (4 M NaOH, for 2 h at 170 degrees C) conditions. The hydroxycinnamic acids released in the hydrolysate were methylated with diazomethane and analysed quantitatively using gas chromatography. Significant portions of ether linkages between hydroxycinnamic acids and lignin were cleaved with DDQ, which suggests that most of the hydroxycinnamic acids were ether-linked at the benzyl position, and not the beta-position, of the lignin side chain as previously claimed. PMID- 11423146 TI - Elucidation of new structures in lignins of CAD- and COMT-deficient plants by NMR. AB - Studying lignin-biosynthetic-pathway mutants and transgenics provides insights into plant responses to perturbations of the lignification system, and enhances our understanding of normal lignification. When enzymes late in the pathway are downregulated, significant changes in the composition and structure of lignin may result. NMR spectroscopy provides powerful diagnostic tools for elucidating structures in the difficult lignin polymer, hinting at the chemical and biochemical changes that have occurred. COMT (caffeic acid O-methyl transferase) downregulation in poplar results in the incorporation of 5-hydroxyconiferyl alcohol into lignins via typical radical coupling reactions, but post-coupling quinone methide internal trapping reactions produce novel benzodioxane units in the lignin. CAD (cinnamyl alcohol dehydrogenase) downregulation results in the incorporation of the hydroxycinnamyl aldehyde monolignol precursors intimately into the polymer. Sinapyl aldehyde cross-couples 8-O-4 with both guaiacyl and syringyl units in the growing polymer, whereas coniferyl aldehyde cross-couples 8 O-4 only with syringyl units, reflecting simple chemical cross-coupling propensities. The incorporation of hydroxycinnamyl aldehyde and 5 hydroxyconiferyl alcohol monomers indicates that these monolignol intermediates are secreted to the cell wall for lignification. The recognition that novel units can incorporate into lignins portends significantly expanded opportunities for engineering the composition and consequent properties of lignin for improved utilization of valuable plant resources. PMID- 11423147 TI - Acidolysis and hot water extraction provide new insights into the composition of the induced "lignin-like" material from squash fruit. AB - Accumulation of "lignin-like" material (L-LM) by plant tissues in response to injury or disease has been observed in a wide variety of plant taxa. The most intensively studied L-LM is that produced by members of the Cucurbitaceae; this material is thought to be an unusual lignin rich in p-coumaryl alcohol derived subunits. Employing acidolysis we found the primary degradation product of L-LM from squash fruit was p-coumaryl aldehyde. These findings conflict with the current concept of L-LM, but would be consistent with L-LM being a polymer derived directly from p-coumaryl aldehyde or a gum containing this compound. Results of hot water extraction support the latter possibility. Further, we report on a simple TLC method useful for rapid qualitative characterization of acidolysis degradation products. PMID- 11423148 TI - A heartwood pigment in Dalbergia cell cultures. AB - In an extensive survey of the genera Baphia, Caesalpinia, Dalbergia, Haematoxylon, and Pterocarpus, we have identified a number of species whose cell cultures accumulated pigments similar to those in heartwood. Thirteen rosewood (Dalbergia) species produced a purple quinonemethide pigment in the callus that was apparently identical between the species. The pigment was first purified from D. retusa cell culture and its structure was elucidated by mass, infrared, and detailed 1H and 13C NMR and NOE spectroscopic studies including 2D experiments (COSY, NOESY, HMQC, and HMBC). Retusapurpurin A (1a) is a C(30) isoflavan of novel skeleton whose formation can be rationalized to occur via regioselective oxidative coupling of an isoflavan to 4,4'-dihydroxy-2'-methoxychalcone. Retusapurpurin A was also isolated from D. parviflora heartwood and cell culture indicating that stress metabolism pathways that are shared with heartwood-type secondary metabolism subpathways are initiated in Dalbergia cell cultures. Therefore, Dalbergia cell cultures afford a good model system for studying heartwood-type metabolic differentiation. PMID- 11423149 TI - Structure and stereochemistry of triflavanoids containing both ether and carbon carbon interflavanyl bonds. AB - The first triflavanoids with both C-C and C-O-C interflavanyl bonds, epioritin (4beta-->6)-epioritin-(4alpha-->4)-epioritin-4beta-ol, epioritin-(4beta-->3) epioritin-(4beta-->6)-epioritin-4beta-ol and epioritin-(4beta-->3)-epioritin (4beta-->6)-epimesquitol-4alpha-ol, were identified in the heartwood of Acacia caffra. The ethereal interflavanyl bond is readily susceptible to reductive cleavage with sodium cyanoborohydride in trifluoroacetic acid/dichloromethane which hence permits the unequivocal assignment of the absolute configuration of constituent flavanyl units. PMID- 11423150 TI - NMR studies of molecular structure in fruit cuticle polyesters. AB - The cuticle of higher plants functions primarily as a protective barrier for the leaves and fruits, controlling microbial attack as well as the diffusion of water and chemicals from the outside environment. Its major chemical constituents are waxes (for waterproofing) and cutin (a structural support polymer). However, the insolubility of cutin has hampered investigations of its covalent structure and domain architecture, which are viewed as essential for the design of crop protection strategies and the development of improved synthetic waterproofing materials. Recently developed strategies designed to meet these investigative challenges include partial depolymerization using enzymatic or chemical reagents and spectroscopic examination of the intact polyesters in a solvent-swelled form. The soluble oligomers from degradative treatments of lime fruit cutin are composed primarily of the expected 10,16-dihydroxyhexadecanoic and 16-hydroxy-10 oxo-hexadecanoic acids; low-temperature HF treatments also reveal sugar units that are covalently attached to the hydroxyfatty acids. Parallel investigations of solvent-swollen cutin using 2D NMR spectroscopy assisted by magic-angle spinning yield well-resolved spectra that permit detailed comparisons to be made among chemical moieties present in the intact biopolymer, the soluble degradation products, and the unreacted solid residue. PMID- 11423155 TI - Development of the visual system of the chick. II. Mechanisms of axonal guidance. AB - The quest to understand axonal guidance mechanisms requires exact and multidisciplinary analyses of axon navigation. This review is the second part of an attempt to synthesise experimental data with theoretical models of the development of the topographic connection of the chick retina with the tectum. The first part included classic ideas from developmental biology and recent achievements on the molecular level in understanding cytodifferentiation and histogenesis [J. Mey, S. Thanos, Development of the visual system of the chick. (I) Cell differentiation and histogenesis, Brain Res. Rev. 32 (2000) 343-379]. The present part deals with the question of how millions of fibres exit from the eye, traverse over several millimetres and spread over the optic tectum to assemble a topographic map, whose precision accounts for the sensory performance of the visual system. The following topics gained special attention in this review. (i) A remarkable conceptual continuity between classic embryology and recent molecular biology has revealed that positional cellular specification precedes and determines the formation of the retinotectal map. (ii) Graded expression of asymmetric genes, transcriptional factors and receptors for signal transduction during early development seem to play a crucial role in determining the spatial identity of neurons within surface areas of retina and optic tectum. (iii) The chemoaffinity hypothesis constitutes the conceptual framework for development of the retinotopic organisation of the primary visual pathway. Studies of repulsive factors in vitro developed the original hypothesis from a theoretical postulate of chemoattraction to an empirically supported concept based on chemorepulsion. (iv) The independent but synchronous development of retina and optic tectum in topo-chronologically corresponding patterns ensures that ingrowing retinal axons encounter receptive target tissue at appropriate locations, and at the time when connections are due to be formed. (v) The growth cones of the retino-fugal axons seem to be guided both by local cues on glial endfeet and within the extracellular matrix. On the molecular level, the ephrins and their receptors have emerged as the most likely candidates for the material substrate of a topographic projection along the anterior-posterior axis of the optic tectum. Yet, since a number of alternative molecules have been proposed for the same function, it remains the challenge for the near future to define the proportional contribution of each one of the individual mechanisms proposed by matching theoretical predictions with the experimental evidence. PMID- 11423156 TI - Social behavior functions and related anatomical characteristics of vasotocin/vasopressin systems in vertebrates. AB - The neuropeptide arginine vasotocin (AVT; non-mammals) and its mammalian homologue, arginine vasopressin (AVP) influence a variety of sex-typical and species-specific behaviors, and provide an integrational neural substrate for the dynamic modulation of those behaviors by endocrine and sensory stimuli. Although AVT/AVP behavioral functions and related anatomical features are increasingly well-known for individual species, ubiquitous species-specificity presents ever increasing challenges for identifying consistent structure-function patterns that are broadly meaningful. Towards this end, we provide a comprehensive review of the available literature on social behavior functions of AVT/AVP and related anatomical characteristics, inclusive of seasonal plasticity, sexual dimorphism, and steroid sensitivity. Based on this foundation, we then advance three major questions which are fundamental to a broad conceptualization of AVT/AVP social behavior functions: (1) Are there sufficient data to suggest that certain peptide functions or anatomical characteristics (neuron, fiber, and receptor distributions) are conserved across the vertebrate classes? (2) Are independently evolved but similar behavior patterns (e.g. similar social structures) supported by convergent modifications of neuropeptide mechanisms, and if so, what mechanisms? (3) How does AVT/AVP influence behavior - by modulation of sensorimotor processes, motivational processes, or both? Hypotheses based upon these questions, rather than those based on individual organisms, should generate comparative data that will foster cross-class comparisons which are at present underrepresented in the available literature. PMID- 11423157 TI - Tau and transgenic animal models. AB - Advances in genetics and transgenic approaches have a continuous impact on our understanding of Alzheimer's disease (AD) and related disorders, especially as aspects of the histopathology and neurodegeneration can be reproduced in animal models. AD is characterized by extracellular Abeta peptide-containing plaques and neurofibrillary aggregates of hyperphosphorylated isoforms of microtubule associated protein tau. A causal link between Abeta production, neurodegeneration and dementia has been established with the identification of familial forms of AD which are linked to mutations in the amyloid precursor protein APP, from which the Abeta peptide is derived by proteolysis. No mutations have been identified in the tau gene in AD until today. Tau filament formation, in the absence of Abeta production, is also a feature of several additional neurodegenerative diseases including progressive supranuclear palsy, corticobasal degeneration, Pick's disease, and frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17). The identification of mutations in the tau gene which are linked to FTDP-17 established that dysfunction of tau can, as well as Abeta formation, lead to neurodegeneration and dementia. In this review, newly recognized cellular functions of tau, and the neuropathology and clinical syndrome of FTDP-17 will be presented, as well as recent advances that have been achieved in studies of transgenic mice expressing tau and AD-related kinases and phosphatases. These models link neurofibrillary lesion formation to neuronal loss, provide an in vivo model in which therapies can be assessed, and may contribute to determine the relationship between Abeta production and tau pathology. PMID- 11423158 TI - Rethinking the role of ceruloplasmin in brain iron metabolism. AB - For more than three decades, it has been widely accepted that ceruloplasmin plays an important role in iron efflux from mammalian cells, including brain cells, via the activity of ferroxidase. However, in light of recent findings, this view might not be completely accurate and the role of ceruloplasmin in brain iron metabolism may need to be re-evaluated. Based on recent studies, we propose in this article that the role of ceruloplasmin in iron uptake by brain neuronal cells might be more important than its role in iron release from the cells. A possible explanation of why the absence of ceruloplasmin induces excessive iron accumulation in neurons in aceruloplasminemia (ceruloplasmin gene mutations) was also discussed. PMID- 11423159 TI - Varieties of human spatial memory: a meta-analysis on the effects of hippocampal lesions. AB - The current meta-analysis included 27 studies on spatial-memory dysfunction in patients with hippocampal damage. Each study was classified on the basis of the task that was used, i.e., maze learning, working memory, object-location memory, or positional memory. The overall results demonstrated impairments on all spatial memory tasks. Clear differences in effect size were found between positional memory on the one hand and maze learning, object-location memory, and working memory on the other hand. Lateralization was found only on maze learning and object-location memory. These findings clearly indicate that specific aspects of spatial memory can be affected in various degrees in patients with hippocampal lesions. Moreover, these results strongly support the notion that the hippocampus is important in the processing of metric positional information, probably in the form of an allocentric cognitive map. PMID- 11423160 TI - Simultaneous modulation of retrieval by dopaminergic D(1), beta-noradrenergic, serotonergic-1A and cholinergic muscarinic receptors in cortical structures of the rat. AB - Retrieval of inhibitory avoidance has been recently shown to require intact glutamate receptors, protein kinases A and C and mitogen-activated protein kinase in the CA1 region of the rat hippocampus and in the entorhinal, posterior parietal and anterior cingulate cortex. These enzymatic activities are known to be modulated by dopamine D(1), beta-noradrenergic, 5HT1A and cholinergic muscarinic receptors. Here we study the effect on retrieval of this task of well known agonists and antagonists of these receptors infused in the same brain cortical regions and into the basolateral amygdala, in rats. The drugs used were SKF38393 (D(1) agonist), noradrenaline, 8-HO-DPAT (5HT1A agonist), oxotremorine (muscarinic agonist), SCH23390 (D(1) antagonist), timolol (beta antagonist), NAN 190 (5HT1A antagonist) and scopolamine (muscarinic antagonist). All were studied at two different dose levels. The localised infusion of SKF38393, noradrenaline, NAN-190 and oxotremorine into any of the cortical structures mentioned 10 min prior to a 24-h retention test session of one-trial step-down inhibitory avoidance enhanced retention test performance. SCH2330, timolol, 8-HO-DPAT and scopolamine hindered retention test performance. In the basolateral amygdala only an enhancing effect of noradrenaline and an inhibitory effect of timolol were seen. Three hours after the infusions, retention test performance returned to normal in all cases. None of the treatments affected locomotion or rearing in an open field or behaviour in the elevated plus maze. Therefore, their effects on retention testing can be attributed to an influence on retrieval. In conclusion, memory retrieval of this apparently simple task requires the participation of CA1, entorhinal, posterior parietal and anterior cingulate cortex, and is strongly modulated by, dopaminergic D(1), beta-noradrenergic, muscarinic cholinergic and 5HT1A receptors in the four areas. The first three types of receptor enhance, and the latter inhibits, retrieval. Only beta-adrenoceptors appears to be involved in the modulation of retrieval of this task by the amygdala. The results bear on the well-known influence of emotion and mood on retrieval, and indicate that this involves many areas of the brain simultaneously. In addition, the results point to similarities and differences between the modulatory mechanisms that affect retrieval and those involved in the consolidation of the same task. PMID- 11423161 TI - Memory disruption in rats with nigral lesions induced by MPTP: a model for early Parkinson's disease amnesia. AB - Intra-nigral administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyrindine (MPTP) caused a lesion in the substantia nigra, compact part (SNc) and a specific loss of dopamine and its metabolites in the striatum of rats. The animals were then tested in the two-way active avoidance task. MPTP-treated animals presented lower learning scores in the training and test sessions, an effect that was not caused by motor impairment or by a decreased sensitivity to footshock since their reaction time to the footshock (unconditioned stimulus - UCS) was not reduced. These lower scores were also not attributable to lower acoustic sensitivity or to a slowing in the association of the sound cue (conditioned stimulus - CS) with the UCS since the reaction time to the CS in the active avoidance response did not differ between MPTP-treated and control groups. Therefore, these results are more properly attributable to an impairment of the memory acquisition and retention processes. In addition, this study is presented as a model of early Parkinson's Disease amnesia and is discussed in terms of the importance of the nigrostriatal pathway to memory acquisition and storage processes. PMID- 11423162 TI - Locomotion towards a goal alters the synchronous firing of neurons recorded simultaneously in the subiculum and nucleus accumbens of rats. AB - Rats were implanted with recording electrodes aimed at the subiculum and nucleus accumbens. They were subsequently placed in a cylindrical environment, where they searched for locations where they would receive rewarding medial forebrain bundle stimulation. At times a tone was sounded, indicating that the reward location was in the center of the environment. Animals quickly learned to switch from random running to goal directed locomotion when the tone was on. To quantify the synchronous firing between simultaneously recorded neurons in the subiculum and nucleus accumbens, a gravitational clustering algorithm was employed. Individual neurons were modeled as particles in N dimensional space. Every spike discharge of the neuron augmented the 'gravitational charge' on its model particle. Synchronous firing between two cells caused their corresponding particles to draw together over time, due to the concurrent appearance of gravitational charge. All pair-wise combinations of cells isolated in subiculum and nucleus accumbens were examined using this algorithm. The firing of nine out of 52 subicular-accumbens cell pairings was significantly more synchronous when the tone was on, and the rat was running towards the central goal. This was also seen for 15 out of 22 subicular-subicular cell pairings. Conversely, only two out of 51 accumbens accumbens pairings displayed significant tone dependent changes in synchronous firing. Thus, synchronous interactions between subiculum and nucleus accumbens occur preferentially when the animal is required to locate a fixed goal in space, i.e., the functional connectivity is altered by the navigational demands of the spatial reward task. PMID- 11423163 TI - Cell activity in the anterior piriform cortex during an olfactory learning in the rat. AB - Several studies have shown that the piriform cortex is involved in learning processes and pyramidal cell activity does not only encode the odour quality but is also related to contextual information about past experience and future action. To study how odour-specific patterns in neuronal activity are established we used an odour discrimination go/no go task with water reinforcement for analysing extracellular single cell activity in anterior piriform cortex in freely moving rats. During conditioning single cells responded to different task events. Of the cells 52% participate in odour sampling and 87% were involved in odour discrimination. More than half of the responses to odours were inhibitory responses. Seventeen percent changed their activity for nose-poke only. The activity of 33% was related to reinforcement. Once established the pattern of reaction to the odour was preserved for several days. It is suggested that the anterior part of the piriform cortex is not involved in odour coding only. However, learning-related plasticity was not observed in this area. PMID- 11423164 TI - A comparison of 129S2/SvHsd and C57BL/6JOlaHsd mice on a test battery assessing sensorimotor, affective and cognitive behaviours: implications for the study of genetically modified mice. AB - Mice from the 129S2/SvHsd (129) and C57BL/6JOlaHsd (C57) strains were assessed for performance on a test battery including sensorimotor, affective and cognitive measures, using established as well as modified paradigms. Marked strain differences were observed in measures of locomotion, motor coordination and anxiety: the 129 mice were typically less active and more anxious. In contrast, the strains did not markedly differ in muscular strength, habituation to an open field or spatial working memory. The data provide baseline parameters of behaviour against which genetically modified lines derived from these particular parental strains can be assessed. PMID- 11423165 TI - Lipopolysaccharide causes deficits in spatial learning in the watermaze but not in BDNF expression in the rat dentate gyrus. AB - We investigated the effects of a single injection and a daily injection of lipopolysaccharide (LPS) on spatial learning and brain-derived neurotrophic factor (BDNF) expression in the rat dentate gyrus. LPS is derived from the cell wall of Gram-negative bacteria and is a potent endotoxin that causes the release of cytokines such as interleukin-1 and tumour necrosis factor. LPS is thought to activate both the neuroimmune and neuroendocrine systems; it also blocks long term potentiation in the hippocampus. Here, we examined the effects of LPS on a form of hippocampal-dependent learning-spatial learning in the water maze. Rats were injected with LPS intraperitoneally (100 microg/kg) and trained in the water maze. The first group of rats were injected on day 1 of training, 4 h prior to learning the water maze task. Groups 2 and 3 were injected daily, again 4 h prior to the water-maze task; group 2 with LPS and group 3 with saline. A number of behavioural variables were recorded by a computerised tracking system for each trial. The behavioural results showed a single injection of LPS (group 1) impaired escape latency in both the acquisition and retention phases of the study, whereas a daily injection of LPS did not significantly impair acquisition or retention. BDNF expression was analysed in the dentate gyrus of all animals. No significant differences in BDNF expression were found between the three groups. PMID- 11423166 TI - Elemental and configural visual discrimination learning following lesions to perirhinal cortex in the rat. AB - Rats were tested in a series of two-choice visual discrimination tasks in a computer-controlled testing apparatus. The discriminations used a range of discriminanda, which varied in complexity. The discriminations included relatively simple form discriminations, more complex form discriminations and discriminations between compound stimuli that shared many features. It was found that rats with perirhinal cortex lesions were unimpaired in all discriminations except those that involved the compound stimuli with overlapping features. Using these stimuli, rats with perirhinal cortex lesions were unimpaired when the stage of learning did not necessitate discriminating stimuli on the basis of more than one feature. However, when efficient performance of the task needed the configuration of more than one feature to be taken into account, perirhinal lesioned rats were impaired. These results are interpreted as revealing the role of the perirhinal cortex in providing multifeature information about the properties of visual objects. PMID- 11423167 TI - Enhancement of immobility induced by repeated phencyclidine injection: association with c-Fos protein in the mouse brain. AB - Immunohistochemistry of c-Fos protein was performed to study changes in neuronal activity in discrete brain areas of mice repeatedly treated with phencyclidine (PCP) showing enhancement of immobility in the forced swimming test, this behavioral change being considered as avolition, which is one of negative symptoms of schizophrenia. Repeated treatment with PCP significantly prolonged immobility time in the forced swimming test, compared with saline treatment. The c-Fos protein expression of mice showing PCP-induced enhancement of immobility was increased in certain brain regions, such as the retrosplenial cortex, pyriform cortices, pontine nuclei, cingulate, frontal cortex and thalamus, compared with that of PCP-treated, non-swimming and saline-treated, swimming groups. These results suggest that increased c-Fos protein is involved in the expression of PCP-induced enhancement of immobility, and c-Fos expression plays a role in negative symptoms-like behavioral changes. PMID- 11423168 TI - Memory deficits in adult rats following postnatal iron administration. AB - Two experiments investigated the effects of Fe(2+), administered postnatally to rat pups on days 10-12, upon tests of memory performance and motor behaviour. In experiment I, Wistar rat pups were administered Fe(2+) at doses of either 2.5, 7.5, 15.0 or 30.0 mg/kg, or vehicle, postnatally, and tested in the open-field at 3 months of age, followed 6 weeks later by testing in the radial arm maze. In the open-field test, only the 30.0 mg/kg Fe(2+) group showed a significantly decreased number of ambulations, but not rearings. In the radial arm maze, all four dose groups, demonstrated deficits in acquisition performance from test days 3 to 5. Retention quotients confirmed the cognitive deficits over all four Fe(2+) groups. In experiment II, rats were administered either 2.5, 7.5 or 22.5 mg Fe(2+) per kg, or vehicle, postnatally, and tested in the inhibitory avoidance (IA) conditioning and retention test at 3 months of age. In the IA conditioning test, groups were either given five 10-min preexposures to the test chamber (preexposed) or simply moved to another cage (non-preexposed). IA retention was blocked in non-preexposed rats administered 7.5 and 22.5 mg Fe(2+) per kg whereas in preexposed rats the 7.5 mg/kg group did not differ from the control (vehicle) group, although the preexposed control group showed significantly better retention than the non-preexposed control group. Postnatal iron administration appears to induce long-lasting detrimental effects upon performance of both appetitively and negatively reinforced tests of memory. Analysis of iron content indicated significant increases in the substantia nigra of the 7.5, 15.0 and 30.0 mg/kg dose groups, but not in the 2.5 mg/kg dose group. Postnatal iron administration appears to induce far-reaching effects upon the performance of certain learned behaviours. PMID- 11423169 TI - Cyamemazine as an anxiolytic drug on the elevated plus maze and light/dark paradigm in mice. AB - Several studies have demonstrated that cyamemazine, a classic antipsychotic compound, possesses anxiolytic properties in humans as well as a potent antagonistic effect on 5-HT(2C) and 5-HT(3) receptors. In this study the level of anxiety of mice was assessed in the light/dark exploration test and the elevated plus maze (EPM) following both acute and chronic administration. Spontaneous locomotor activity was measured using a photoelectric actimeter. Acute or chronic administration of cyamemazine dramatically decreases the spontaneous locomotor activity of mice at the dose of 1 mg/kg in comparison with the control group. In the light/dark exploration test, cyamemazine (0.375 mg/kg) only demonstrated anxiolytic-like activity following acute administration. In the elevated plus maze (EPM), cyamemazine did not induce any anxiolytic like effects after acute administration. However, after chronic administration, cyamemazine at doses of 0.25, 0.375, 0.5 and 1 mg/kg significantly increased the time spent in the open arms. The number of open arm entries was also increased at 0.25 and 0.5 mg/kg. Various serotonergic ligands were then used to examine the role of the various receptors in mediating the effects of cyamemazine in the EPM. Concerning the 5 HT(2) ligands DOI and mCPP antagonised the effects of cyamemazine and N-desmethyl clozapine potentiated the effects. For 2-methyl-5-HT and ondansetron, the 5-HT(3) receptor ligands did not seem to have any effect. It appears that the 5-HT(2C) receptors are more implicated in the function of mediating the anxiolytic effect of cyamemazine in the EPM. The discrepancy of results obtained in the tests, following acute or chronic administration could be the result of the combined activity of dopamine D(2) receptor antagonism with antagonism of 5-HT(2C) and 5 HT(3) receptors. PMID- 11423170 TI - Spatial learning in the guinea pig: cued versus non-cued learning, sex differences, and comparison with rats. AB - This paper provides the first report of spatial learning in guinea pigs using the Morris water maze (MWM). Male and female guinea pigs were trained for 5 consecutive days (8 trials/day; acquisition phase) in either the visible (cued) or the hidden (non-cued) platform version of the MWM. In both tests, guinea pigs learned to navigate to the escape platform, as indicated by a decrease in escape latency over the 5 training days. There were no sex differences in either test version. A comparison of guinea pigs and male Wistar rats showed that performance during acquisition training was not different for the two species in the visible platform test, but rats performed better during the early training days in the hidden platform test. A retention test (probe trial) was given 5 days after the last acquisition training day. Again, there was no sex difference, and no difference between guinea pigs and rats. Finally, acquisition of a new escape response to a shifted platform location was equivalent for rats and guinea pigs of both sexes. These results demonstrate that guinea pigs show robust cued and non-cued learning in the MWM. Both acquisition and retention performance in guinea pigs is similar to that in rats, even though rats appear to have a slight advantage in the acquisition of non-cued, spatial information in this test. We conclude that the MWM provides a valuable paradigm to assess behavior and learning/memory in the guinea pig. PMID- 11423171 TI - Permissible and immunogenic HLA-A mismatches: cytotoxic T-cell precursor frequencies reflect graft survival data. AB - Analysis of the in vivo immunogenicity of single HLA mismatches, in the context of a patient's own human leukocyte antigen (HLA) phenotype, has been used to define permissible and immunogenic HLA mismatches. Kidney graft survival in the case of permissible mismatches was similar to that of completely HLA matched combinations, whereas immunogenic mismatches lead to a significantly poorer graft survival. The present study tested whether such permissible and immunogenic HLA mismatches are reflected in the in vitro cytotoxic T-lymphocyte (CTL) allorepertoire. Limiting dilution experiments were performed to analyze the number of precursor CTL directed against individual HLA class I antigens. In general, the frequency of CTLp directed against permissible HLA-A antigens (n = 70, mean frequency 27 CTLp per million peripheral blood lymphocytes [PBL]) was found to be significantly lower compared with the CTLp directed against immunogenic HLA-A antigens (n = 73, mean frequency 59 CTLp per million PBL). The difference was found both in healthy individuals and a population of renal transplant candidates. These results were confirmed by a retrospective analysis of CTLp frequencies performed between partly mismatched unrelated bone marrow donors and their potential recipients. In conclusion, on the population level the permissible and immunogenic HLA-A mismatches are indeed reflected in the CTL allorepertoire. However, due to the big overlap of the CTLp frequencies in these populations, the permissible or immunogenic nature of a mismatch for a particular patient should be determined on an individual basis. PMID- 11423172 TI - In vivo stability of human chemokine and chemokine receptor expression. AB - Cross-sectional analyses of human PBMC, plasma, and tissue have reported altered chemokine and/or chemokine receptor expression in several inflammatory diseases. Interpretation of such studies is difficult without data on the in vivo stability of such parameters. Using four color flow cytometry, we longitudinally followed CXCR3, CCR5 (Th1-associated), and CCR3 (Th2-associated) expression within CD4+/CD45RO+ and CD8+/CD45RO+ T cell populations in peripheral blood of healthy individuals over a 21 day period. In parallel, we quantified plasma levels of IP 10, Mig, eotaxin and TARC. Chemokine and receptor expression differed markedly between subjects but was highly stable, varying by <5% within individuals. Differences in chemokine receptor expression between subjects were markedly altered when quantified as absolute cell numbers rather than frequencies. Finally, CCR3 expression by CD4+/CD45RO+ T cells was positively correlated with plasma levels of its ligand, eotaxin, whereas strong negative correlations were evident between CXCR3 expression and IP-10 or Mig. These data demonstrate longitudinal stability of chemokine receptor and ligand expression among healthy individuals; reveal that both frequency and absolute cell count analysis is essential for accurate assessment of chemokine receptor expression; and identify inverse relationships between type 1 and type 2 immunity-associated receptors and their ligands in vivo. PMID- 11423173 TI - The Fc receptor for IgG (Fc gamma RII; CD32) on human neonatal B lymphocytes. AB - B cells express an Fc receptor for IgG (FcgammaRII; CD32) which is involved in feedback inhibition of antibody production. Engagement of FcgammaRII during ligation of the antigen receptor provides an inhibitory signal. FcgammaRII exists as several isoforms, with FcgammaRIIb (which carries an immunoreceptor tyrosine based inhibition motif; ITIM) being predominant form on adult B cells. The inhibitory role of FcgammaRIIb may be unhelpful to the infant, since primary exposure to infectious agents is likely to be in the presence of maternal IgG. We hypothesized that neonatal B cells would be less susceptible to feedback inhibition by antibody, either through the expression of activation-competent FcgammaRII isoforms (FcgammaRIIa and FcgammaRIIc) or through reduced expression of the inhibitory FcgammaRIIb isoforms. Cord and adult B cells were examined for expression of FcgammaRII isoforms using monoclonal antibodies and RT-PCR. In vitro assays were performed to assess susceptibility of cord and adult cells to FcgammaRII-mediated suppression. Although there is no phenotypic difference in FcgammaRII expression (FcgammaRIIb predominating on both adult and cord B cells), FcgammaRIIb is expressed at lower levels on cord cells. This quantitative difference in FcgammaRIIb expression may explain the reduced susceptibility of cord B cells to antibody-mediated inhibition observed in these experiments. PMID- 11423174 TI - Beryllium binding to HLA-DP molecule carrying the marker of susceptibility to berylliosis glutamate beta 69. AB - Berylliosis is a chronic granulomatous disorder caused by inhalation of Be dusts that is driven by the accumulation of Be-specific CD4+ Th1-cells at disease sites. Susceptibility to berylliosis has been associated with the supratypic variant of HLA-DP gene coding for glutamate at position beta69 (HLA-DPbetaGlu69). The aim of this study was to test the hypothesis that the HLA-DPbetaGlu69 residue plays a role in the interaction with Be. To this end, soluble HLA-DP2 molecule (carrying betaGlu69) and its mutated form carrying lysine at position beta69 (HLA DP2Lys69) were produced in Drosophila melanogaster and then used in a Be binding assays. BeSO4 (1-1000 microM) was used to compete for the binding of the biotinilated invariant chain-derived peptide CLIP (50 microM). BeSO4 was capable of compete out biotin-CLIP binding from the HLA-DP2 (IC50%: 4.5 microM of BeSO4 at pH 5.0 and 5.5 microM of BeSO4 at pH 7.5), but not from the HLA-DP2Lys69 molecule (IC50%: 480 microM of BeSO4 at pH 5.0 and 220 microM of BeSO4 at pH 7.5). Moreover, the binding of NFLD.M60, a MoAb recognizing an epitope in the HLA DP peptide binding region, to the HLA-DP2, but not to the HLA-DP2Lys69 soluble molecules was inhibited BeSO4. NFLD.M60 binding to HLA-DP2, but not to HLA DP2Lys69 stably transfected murine cells was also inhibited by Be both at pH 5.0 and at pH 7.5. The data indicate a direct interaction of Be with the HLA-DPGlu69 molecule, in the absence of antigen processing. PMID- 11423175 TI - CD28+ intraepithelial lymphocytes with long telomeres are recruited within the inflamed ileal mucosa in Crohn disease. AB - Crohn disease is a chronic inflammatory bowel disease that involves all the intestine but predominantly alters the ileum. The disease largely depends on T cells, but the biologic role of intestinal intraepithelial lymphocytes (IEL) in transmural inflammation remains poorly characterized. To address this issue, a comparison of IEL and lamina propria lymphocytes (LPL) isolated from the uninvolved and the inflamed ileal mucosa of Crohn disease patients was performed. More CD8+ IEL (26% versus 8%) from the inflamed ileal mucosa expressed the CD28 receptor and the CD11a integrin than IEL from the uninvolved ileal mucosa, which were mostly CD28-. IEL had longer telomeres in the inflamed than in the uninvolved areas and a TCR Vbeta repertoire more similar to circulating T cells, suggesting that the increased proportion of CD28+ TCRalphabeta+ IEL within the inflamed mucosa is more likely due to recruited lymphocytes from the periphery that populate the epithelial layer than to the acquisition of the CD28 molecule by activated resident lymphocytes. In the uninvolved ileal mucosa, IEL from Crohn disease patients had shorter telomeric lengths than IEL from control patients, suggesting that they have been chronically stimulated. Such perturbation of the IEL population within the ileal mucosa could contribute to the inflammation in Crohn disease. PMID- 11423176 TI - Crohn disease: susceptibility and disease heterogeneity revealed by HLA genotyping. AB - Predisposition to Crohn disease (CD) seems to be genetically determined but, though several reports on the matter, the association between HLA antigens and the disease is still controversial. PCR-SSP high resolution typing in 107 CD patients, and in subgroups selected according to clinical features, showed a positive association with the rare haplotype DRB1*07, DQB1*0303 both in the overall patients (p = 0.002; pc = ns) and in the subgroup of nonfistulized patients (p = 0.0008; pc = 0.032). Moreover, the protective role of the haplotype DRB1*03, DQB1*0201 (p = 0.029) was confirmed also in Italian patients, whereas no strong association with HLA class I alleles has been found. In addition, variability of the HLA alleles frequency in CD subgroups was observed, supporting the hypothesis of a genetic heterogeneity of the disease and suggesting that HLA alleles distribution in selected groups may allow to identify patients with probably different prognosis or associated complications. PMID- 11423177 TI - A genetically determined high setting of TNF-alpha influences immunologic parameters of HLA-B8,DR3 positive subjects: implications for autoimmunity. AB - The 8.1 ancestral haplotype (AH) is a common Caucasoid haplotype carried by most people who type for HLA-B8,DR3. It seems unique in its association with a wide range of immunopathologic diseases. Healthy subjects bearing this haplotype demonstrate several alterations of immune response. This article will focus on the identification of the mechanism(s) of disease susceptibility of 8.1 AH. In 13 carriers of 8.1 AH, and 43 negative patients, enzyme immune assays serum levels of tumor necrosis factor (TNF)-alpha, soluble endothelial leukocyte adhesion molecule-1 (sELAM-1), cortisol, and interleukin(IL)-10 were determined. In addition, quantification of cytokine produced in vitro after mitogen stimulation was studied, and all subjects were genotyped for alleles at -592, -819, and -1082 nucleotides of IL-10 gene 5' flanking region, which is known to control IL-10 production. Results revealed that 8.1 AH is associated with a high in vivo and in vitro production of TNF-alpha, which in turn seems responsible for increased serum levels of sELAM-1, cortisol, and IL-10. On the contrary, in vitro production of IL-10 is not increased in these patients and there are no differences in allele promoter frequencies between the two groups that might explain the differences in IL-10 serum values. Thus, serum values seem to be the result of the effects of increased serum levels of TNF-alpha and cortisol. In conclusion, the increased spontaneous release of TNF-alpha, which modifies a certain number of immunologic parameters, may be the most characterizing feature of 8.1 AH. The consequent modification of the immunologic scenario might be involved in the predisposition to the impressive number of diseases and the changes in immune response observed in the patients studied. PMID- 11423178 TI - Immune related genetic polymorphisms and schizophrenia among the Chinese. AB - Genetic association studies were conducted among two independent cohorts of Chinese ethnicity. The samples consisted of cases and unrelated controls, ascertained from Guangzhou, China, and Singapore. The studies were prompted by our earlier report of an association between schizophrenia and HLA DQB1 alleles (HLA DQB1*0602 and HLA DQB1*0303) in the Singapore sample. Polymorphisms of HLA DQB1 and flanking markers on chromosome 6p21.3 were investigated in the first part of the study. A significant negative association with HLA DQB1*0402 was detected in the Guangzhou sample (Odds ratio, OR 0.26, 95% confidence intervals, CI 0.1, 0.6; p < 0.02, corrected for multiple comparisons). Additional analysis of the Guangzhou and Singapore samples revealed associations at three other anonymous markers flanking HLA DQB1. In the second part of the study, three polymorphisms at the Interleukin-1 gene cluster (IL-1, chromosome 2q13-q21) were investigated in both cohorts, since associations with schizophrenia have been reported in another sample. Persuasive evidence for an association at IL-1 was not detected in either sample. Our results suggest a susceptibility locus for schizophrenia in the HLA region among the Chinese, but further clarification is necessary. PMID- 11423179 TI - HLA class I and class II are both associated with the genetic predisposition to primary Sjogren syndrome. AB - Primary Sjogren syndrome (pSS) is an autoimmune disease characterized by progressive destruction of the exocrine glands leading to mucosal and conjunctival dryness. It is marked by lymphocytic infiltration of the glands and the accumulation of several types of autoantibodies such as rheumatoid factor (RF), antinuclear, anti-SS-A (anti-Ro) and anti-SS-B (anti-LA) autoantibodies. The susceptibility to pSS and/or the presence of SS-A/SS-B autoantibodies in pSS patients is associated with DRB1*03-DQB1*02 and DRB1*02-DQB1*06 haplotypes, whereas no associations have been described with any HLA class I allele. To define the impact of HLA class I alleles in predisposition to pSS, 46 patients responding to the European criteria and 222 healthy unrelated Caucasians were analyzed for their HLA class I and class II haplotypes. Our results confirm the association of the DRB1*03-DQB1*02 haplotype with SS-A/SS-B autoantibodies positive pSS and demonstrate a significant association of the HLA-A24 with the disease. Moreover, HLA-A24 is more often associated with DRB1*11-DQB1*0301 and/or DRB1*0301-DQB1*02 in pSS patients than in the controls. The novel association of HLA class I alleles with susceptibility to pSS provides new insights to the genetic predisposition to this disease and subsequently to its physiopathology. PMID- 11423180 TI - A unique murine monoclonal antibody recognizing HLA-B53, B37, B51, B52, +/-B44. AB - Monoclonal antibodies have played an important role in studying the biochemistry of the HLA-Class I molecules. Some murine anti-HLA mAbs can identify configurations of HLA epitopes that have never been reported in human allosera. One of these configurations is identified by an IgM mAb designated as: BHA-1441. This antibody was produced using a lymphoblastoid cell line typed as: A*02, A*25; B*38, B*4402/4405; C*0501, C*07, BW4, as the immunogen. A lymphocytotoxicity test of this mAb over a panel of 109 frozen, 452 fresh and, later, 44 DNA typed T cells revealed its specificity as B53, 37, 51, 52, +/- 44. All of the antigens recognized by this mAb share the Bw4 motif at positions 81-83, except for the HLA B37, which shares only 82L and 83R. Furthermore, while B37 and B44 cross-react due to the aspartic acid (D) substitution at position 156, the reactivity with B53, B5 (51,52), B37 and 60% of B44 cells, makes it unlikely that the target epitope could be due only to the primary amino-acid sequence. The antibody binding site might involve changes in tertiary structure and peptides bound by the MHC. BHA-1441 is an interesting tool to study and type the HLA-B53 antigen and its cross-reactive epitopes. PMID- 11423181 TI - Improved methods for the isolation and purification of porcine islets. AB - Recent progress in human islet transplantation demonstrates the feasibility of using purified human islets for treatment of type 1 diabetes mellitus; however, a shortage of human pancreata remains a major obstacle. This report describes methods to isolate porcine islets using a modification of the automated chamber method. The pancreata from 2-year-old sows were trimmed and injected intraductally with Sevac, Sigma, or Liberase PI collagenase. The pancreata was placed in the chamber, shaken, and recirculated at 70 ml/min until an adequate number of islets were liberated. The digest was centrifuged and the pellets pooled with University of Wisconsin Solution + 10% horse serum and incubated at 4 degrees C for 1 h. The islets were purified using a continuous gradient of Hypaque Euroficoll on a refrigerated COBE 2991. The islets were collected in fractions, assessed for purity, sized, and then suspended in Medium 199. Collagenase preparations obtained from Sevac (2919 islet equivalents [IE]/g), Sigma (2543 IE/g), and Liberase PI (2901 IE/g) gave similar results with 94%-95% purity. In summary, we report a successful method for efficient isolation and purification of porcine islets, yielding nearly 3000 IE/gm, with different collagenase products. PMID- 11423182 TI - Detection of IgG antibodies to Neospora caninum and Toxoplasma gondii in dogs examined in a veterinary hospital from Brazil. AB - A total of 163 dogs with neuromuscular, respiratory and/or gastrointestinal disorders, was admitted at the Veterinary Hospital, Federal University of Uberlandia, Brazil, and submitted to serology for Toxoplasma gondii and Neospora caninum. Assays for T. gondii included indirect haemagglutination (IHA), indirect fluorescent antibody (IFAT-Tg), immunoenzymatic (ELISA), and immunoblotting (IB Tg). Assays for N. caninum included IFAT-Nc and immunoprecipitation (IP-Nc). Based on concordant results by three serological tests (IHA, IFAT-Tg and ELISA) for T. gondii, and divergent results further confirmed by IB-Tg for reactivity to TgSAG1, the 163 sera were divided into two groups: 59 (36%) Tg-seropositive samples and 104 (64%) Tg-seronegative samples. Antibodies to Neospora were detected in 11 (6.7%) out of 163 analyzed dog sera, with 5 (3.1%) samples reactive to both parasites (Tg+/Nc+), and 6 (3.7%) reactive only to Neospora (Tg /Nc+). Antibodies only to T. gondii were found in 54 (33%) samples. Among the 11 Neospora-positive sera analyzed by IB-Tg, the five sera Tg+/Nc+ showed strong reactivity to Toxoplasma antigens, especially to TgSAG1 (p30). No reactivity was observed to TgSAG1 in the six samples Tg-/Nc+. By IP-Nc, two highly immunodominant antigens (29 and 35kDa proteins) were recognized by all 11 IFAT-Nc positive sera. Our results suggest that the infection by N. caninum can be concomitantly present in dogs from this area, although less common, and therefore should be considered in the differential clinical diagnosis with T. gondii in dogs presenting neuromuscular, respiratory and/or gastrointestinal disorders. PMID- 11423183 TI - A randomised, blinded, placebo-controlled clinical trial with allopurinol in canine leishmaniosis. AB - A total of 45 non-uremic dogs, with clinical signs indicating leishmaniosis, entered the study. Diagnosis was confirmed by indirect immunofluorescence assay (IFA) on serum and polymerase chain reaction (PCR) on bone marrow samples. The dogs were randomly allocated into Group A (n=37) that received allopurinol (10mg/kg B.W., per os, twice daily) for 4 consecutive months, and Group B (n=8) that were placebo-treated. Clinical signs were scored just before and at monthly intervals throughout the study period, in a blinded and independent fashion. Complete blood count, serum biochemistry profile, urinalysis, lymph node and bone marrow parasitology, IFA and enzyme-linked immunosorbent assay (ELISA) serology and bone marrow PCR were carried out at the beginning and at the end of the trial. A total of three Group A and one Group B dogs died of end stage kidney disease that developed during the trial. In Group A animals that endured the trial there was a significant improvement in the general body condition, conjunctivitis, peripheral lymphadenopathy, splenomegaly, masticatory muscle atrophy, ulcerative stomatitis, epistaxis, exfoliative dermatitis, cutaneous ulcerations, blepharitis and nasodigital hyperkeratosis. The same observation was made for anemia, lymphopenia, hyperproteinemia, hyperglobulinemia, hyperphosphatemia, increased alkaline phosphatase activity and the low albumin/globulin ratio. By contrast, no improvement of any kind was seen in Group B dogs. Lymph node and bone marrow parasite numbers were significantly decreased in Group A animals. In Group B, that occurred only in the lymph nodes. Apart from remission of clinical signs and restoration to normal of clinicopathological abnormalities, allopurinol did not eliminate Leishmania organisms, as the PCR result on bone marrow was still positive in all the dogs that finished the trial. PMID- 11423184 TI - Ovine Echinococcus granulosus transmission dynamics in the province of Rio Negro, Argentina, 1980-1999. AB - In this work, the impact of a hydatidosis control programme for dogs on the intermediate ovine host was specifically analysed to determine the levels of prevalence achieved and the dynamics of parasite transmission, as well as to evaluate the quality of diagnostic systems in slaughterhouses. A field study was conducted in four slaughterhouses (Valcheta, Los Menucos, Jacobacci, Bariloche) that process animals coming from all the departments within the work area. The control programme for dogs entailed treating dogs with 5 mg/kg praziquantel at 2 month intervals for 20 years. Sample size was determined with a 10% error margin and a 95% significance level. Harvested viscera (liver, lungs, kidneys) were preserved in 5% formaldehyde and sent to the laboratory for diagnostic confirmation of both positive and negative specimens. The 61% initial prevalence dropped to 18.3% at the end of the 10-year period, observed differences proving significant (Chi-square=15.454, P=0.00). There were statistically significant increases in infection prevalence with age (Pearson's Chi-square=133.61, P=0.00). Overall, 37.2% of hydatidosis cases diagnosed in slaughterhouses were considered non-hydatid by histological study. On the other hand, 1.1% of those diagnosed as healthy were found to be infected with hydatidosis. The number of hydatid cysts per animal increased with age: 0.04 in lambs and 1.22 in adults (linear regression equation, -0.0539+0.0127 x age), whereas the average for the whole period was 3.7% in lambs and 20.5% in adults. Viability studies indicated that 63.8% of parasitised animals had viable cysts, out of which 53.3% were fertile. Diagnosis of infection in sheep made by means of an adjusted statistical design and with histological confirmation of the presumptive diagnosis made in slaughterhouses demonstrated the flaws of the official systems for epidemic surveillance of hydatidosis. However, there was no overall significant difference in slaughterhouse and laboratory data.Ro was 0.53 and could imply a decrease in reproductive capacity of the parasite and steady state extinction. PMID- 11423185 TI - Epidemiological observations and heterosis analysis of gastrointestinal nematode parasitism in Suffolk, Gulf Coast Native, and crossbred lambs. AB - A crossbreeding program was conducted to evaluate the resistant status to gastrointestinal nematode parasite infection in crossbred (F1) lambs and their Suffolk and Gulf Coast Native (Native) breed counterparts. A total of 253 lambs were included in the study over 2 years. Fecal egg count (FEC) and blood packed cell volume (PCV) of 18-20 monitor lambs of each breed group in each year were collected every other week from birth to 24-30 weeks of age. The FEC and PCV of all lambs were determined at weaning (11-12 weeks of age) and at three subsequent times when anthelmintic treatment was administered. Nematode counts of wether lambs were obtained at 30 weeks of age in the first year. The epidemiological patterns of FEC and PCV of each breed group were similar in both years. The Suffolk group consistently showed the highest FEC and the lowest PCV. Conversely, the Native group had the lowest FEC and highest PCV. For the most part, all parameters for the F1 group fell intermediate to the Native and Suffolk groups. Nematode count followed the same pattern. Heterosis analysis showed that FEC, PCV, nematode count and weight gain of the F1 group favored the Native group. These results suggested that crossbreeding Suffolk to Native sheep may be a suitable way to produce lambs with improved resistance to gastrointestinal nematode infection, but production may be compromised. PMID- 11423186 TI - Preliminary observations on the potential of gut membrane proteins of Haemonchus contortus as candidate vaccine antigens in sheep on naturally infected pasture. AB - In a trial lasting 11 months in South Africa, faecal egg counts and haematocrits of sheep vaccinated with gut membrane proteins of adult Haemonchus contortus were compared with unvaccinated controls grazing pasture contaminated with the parasite. Vaccination reduced egg output by >82% on average during one 4 month period of the trial and simultaneously significantly reduced the degree of anaemia and deaths due to haemonchosis. Although vaccine immunity was not sufficiently long lasting to prevent a surge in egg output which occurred after the onset of a period of irrigation, re-vaccinating the sheep at this point cleared their newly acquired infection and rapidly restored protection to approximately the level observed beforehand. It was clear that a vaccine based on parasite gut membrane proteins could offer substantial benefits in the control of natural haemonchosis. PMID- 11423187 TI - Resistance of leopard tortoises and helmeted guineafowl to Cowdria ruminantium infection (heartwater). AB - Experimental infection trials were conducted to investigate susceptibility of leopard tortoises (Geochelone pardalis) and helmeted guineafowl (Numida meleagris) to infection with Cowdria ruminantium, the causative agent of heartwater, a tickborne disease of domestic and wild ruminants. Ten guineafowl were inoculated intravenously with a virulent dose of C. ruminantium derived from bovine endothelial cell cultures, and four leopard tortoises were exposed to C. ruminantium infection by the feeding of infected Amblyomma hebraeum ticks. Uninfected A. hebraeum ticks (on both tortoises and guineafowl) and Amblyomma marmoreum ticks (on tortoises only) were fed on the animals during weeks 2 and 3 post-exposure in an attempt to detect infection. These ticks were analyzed for C. ruminantium infection by xenodiagnosis and with the C. ruminantium-specific pCS20 polymerase chain reaction (PCR) assay. Attempts to detect infection in ticks fed on either species were negative by both tests. These results suggest that leopard tortoises and helmeted guineafowl are refractory to C. ruminantium infection and, therefore, are unlikely to be capable of introducing heartwater directly into new areas. However, leopard tortoises are efficient hosts of A. marmoreum and A. hebraeum and are likely to be important epidemiologically in the transport and maintenance of these tick vector species. PMID- 11423188 TI - Evaluation of the impact of parasite control with the IVOMEC SR Bolus given at breeding age on first lactation yield in Holstein heifers. AB - This field trial was conducted to test the effect of treatment with a sustained release ivermectin bolus on first lactation milk production (projected to 305 days) and milk composition (percent fat and protein) in breeding-aged Holstein heifers. From 12 herds, 96 heifers were randomly allocated at turnout to treatment with a commercially available ivermectin bolus (n=49) or untreated control (n=47) groups. Animals were weighed at turnout, mid-season, and at the end of the grazing season. Fecal samples were taken and nematode eggs counted. Projected 305 days milk production, fat and protein content were retrieved from PATLQ reports (Quebec DHI records) for heifers that calved during the subsequent 10 months. The heifers had a mean weight of 409 and 404 kg at turnout, respectively, for control and treated groups. Nematode egg excretion was relatively low (mean of 10 egg/g (EPG) for controls at housing) throughout the pasture season. The average daily gain (ADG) and total weight gain (TWG) over the entire pasture season was found to be higher in treated heifers compared to controls (ADG: difference=0.08 kg per day, P=0.010; TWG: difference=12.82 kg, P=0.013). Taking into account a possible farm effect, the 305 days milk production for IVOMEC SR bolus-treated heifers was significantly higher than for untreated animals (mean+/-S.D.; 348+/-476 kg; t-test=2.31, P<0.05). Overall, 8 of the 10 remaining farms in the study demonstrated a positive milk production difference in treated (n=34) versus control heifers (n=33). The results of this study suggest that preventive treatment of breeding-aged, grazing dairy heifers with a sustained-release ivermectin bolus provides a significant weight-gain which may translate to a significant milk production advantage, even in situations with commercial utilization of moderately contaminated pastures. PMID- 11423189 TI - Re-evaluation of ivermectin efficacy against equine gastrointestinal parasites. AB - Two trials were conducted to confirm the efficacy of ivermectin paste against endoparasites of horses. In these trials, 20 ponies were treated with ivermectin oral paste at 200 mcg x kg body weight once on Day 0, and 20 ponies served as unmedicated controls. The animals carried naturally acquired parasite infections as confirmed by pretrial fecal examination. The animals were necropsied for worm recovery on Days 14, 15 or 16. Parasites recovered were identified to species. Horses treated with ivermectin had significantly (P<0.05) fewer (>99.0% reduction) adult small strongyles (Coronocyclus spp including C. coronatus, C. labiatus, C. labratus; Cyathostomum spp including C. catinatum, C. pateratum; Cylicocyclus spp including C. ashworthi, C. elongatus, C. insigne, C. leptostomum, C. nassatus, C. radiatus; Cylicodontophorus bicoronatus; Cylicostephanus spp including C. asymetricus, C. bidentatus, C. calicatus, C. goldi, C. longibursatus, C. minutus; Gyalocephalus capitatus; Parapoteriostomum spp including P. euproctus, P. mettami; Petrovinema poculatum; Poteriostomum spp including P. imparidentatum, P. ratzii) and adult large strongyles (Strongylus edentatus, S. vulgaris; Triodontophorus spp including T. brevicauda, T. serratus; Craterostomum acuticaudatum) than the controls. Ivermectin was also highly effective (94% to >99%, P<0.05-0.01) against Gasterophilus intestinalis larvae, Habronema spp., Oxyuris equi, Parascaris equorum. The data from these two trials confirm that ivermectin paste administered to horses orally at 200mcg x kg(-1) continues to be highly effective for treatment and control of a broad range of small and large strongyle species as well as other species of gastrointestinal parasites. PMID- 11423190 TI - Clostridium perfringens beta toxin and Clostridium septicum alpha toxin: their mechanisms and possible role in pathogenesis. AB - The Clostridium septicum alpha toxin and the Clostridium perfringens beta toxin are examples of pore-forming toxins that exhibit several different features. The cell types that are targeted by these toxins reflect the effect these toxins have on the host during infection with either organism. Alpha toxin elicits a rapid shock-like syndrome, whereas beta toxin appears to induce a variety of neurological effects. The effects of the purified toxins appear to mimic some of the features of the animal and human diseases caused by C. septicum and C. perfringens. This review, examines the current state of knowledge for the cytolytic mechanism, role in pathogenesis and structure of these two toxins. PMID- 11423191 TI - Antigenic properties and experimental transmission to several fish species of a marine birnavirus isolated from sole (Solea senegalensis). AB - A cross-neutralization test was used to study the antigenic relationship of an aquabirnavirus isolated from sole (Solea senegalensis), named solevirus, and several infectious pancreatic necrosis virus (IPNV) strains. Solevirus was antigenically similar to IPNV strain Sp. Transmission of the solevirus to other fish species has been determined by inoculation to freshwater and marine fish species (two salmonids and gilt-head seabream). A higher pathogenicity was obtained for the marine fish species, although solevirus caused an asymptomatic infection in all species tested, as demonstrated by the detection of viral RNA and of viral antigens in fish leucocytes, respectively, using polymerase chain reaction (PCR) and flow cytometry (FC). PMID- 11423192 TI - Molecular analysis of field strains of Mycoplasma capricolum subspecies capripneumoniae and Mycoplasma mycoides subspecies mycoides, small colony type isolated from goats in Tanzania. AB - A molecular analysis of strains of Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae) and Mycoplasma mycoides subsp. mycoides, small colony type (M. mycoides SC) isolated from goats was performed using the amplified fragment length polymorphism (AFLP) and pulsed-field gel electrophoresis (PFGE) fingerprinting techniques. Among the 11 field strains of M. capripneumoniae from Tanzanian goats, two AFLP patterns were demonstrated, with 10 of the strains showing indistinguishable patterns. Five Kenyan strains of M. capripneumoniae produced three AFLP patterns, with two of them being indistinguishable from the 10 identical Tanzanian and one Ugandan strain (M74/93) isolated from sheep. The AFLP pattern of the type strain (F38(T)) was identical to two Kenyan strains (Baringo and G183/82). On PFGE analysis, all the examined M. capripneumoniae strains exhibited identical PFGE profiles.Five field strains of M. mycoides SC isolated from goats displayed identical AFLP patterns except for one strain which differed from others at only one position. The AFLP pattern of the type strain of M. mycoides SC (PG1(T)) was different from the field strains. The five field strains of M. mycoides SC produced identical PFGE profiles, which were, however, different from the type strain. The AFLP and PFGE profiles of M. mycoides SC strains from goats were identical to those of six strains isolated from cattle affected with contagious bovine pleuropneumonia (CBPP) in the same areas. The results of this study suggest a close epidemiological linkage between strains of M. capripneumoniae and between M. mycoides SC type, respectively, isolated from goats in Tanzania. PMID- 11423193 TI - PCR detection and prevalence of alpha-, beta-, beta 2-, epsilon-, iota- and enterotoxin genes in Clostridium perfringens isolated from lambs with clostridial dysentery. AB - Clostridium perfringens isolated from lambs with dysentery (n=117) were analysed by a DNA amplification technique, the polymerase chain reaction (PCR), in order to determine the prevalence of the alpha-, beta-, beta 2-, epsilon-, iota- and enterotoxin genes. The most prevalent toxin type of C. perfringens found was type B, containing the alpha-, beta-, and epsilon-toxin genes, representing 46% of the cases with clostridial dysentery. C. perfringens type C containing the alpha-, and beta-toxin genes was isolated in 20% and type D, which is characterized by the alpha- and epsilon-toxin genes, was isolated in 28% of all isolates. The recently discovered, not yet assigned beta 2-toxigenic type of C. perfringens was represented in 6% of all isolates. No C. perfringens type A containing the alpha toxin alone and no type E, which harbours the ADP-ribosylating iota-toxin, were found in the diseased animals. None of the samples contained the enterotoxin gene. Only one type of C. perfringens was found in a given herd, revealing the epidemiological use of PCR toxin gene typing of C. perfringens. The animals originated from 79 different herds with sizes ranging from 30 to 250 animals, bred in the area of northern Greece. PMID- 11423194 TI - Influence of enrichment media and application of a PCR based method to detect Salmonella in poultry industry products and clinical samples. AB - To attempt the rapid detection of Salmonella enterica, we have coupled a culture procedure with PCR amplification of the genus-specific invE/invA genes. The method was applied to different kinds of samples from the poultry industry and evaluated by using hydrolyzed feather meal, meat meal, litter and viscera, all experimentally inoculated with a known number of Salmonella followed by cultivation in selenite--cystine broth prior to the PCR reaction. The expected 457bp specific DNA fragment could be amplified from dilutions containing as few as 5.7CFU, indicating that the PCR technique can be successfully coupled with culture in an enrichment broth to distinguish Salmonella species from other enteric bacteria present in samples from the poultry industry. Tetrathionate broth proved to be a much better enrichment media compared to selenite-cystine when the presence of Salmonella was evaluated by PCR in 1-day-old chicks experimentally infected with known numbers of Salmonella. Samples included cecal tonsils and viscera, collected at 48h and 7 days postinfection. The PCR technique was more sensitive in detecting infected animals than the standard microbiological procedure, which detected only 47% of all PCR positive samples. PMID- 11423195 TI - Comparison of an indirect ELISA with the Brucella milk ring test for detection of antibodies to Brucella abortus in bulk milk samples. AB - An indirect enzyme-linked immunosorbent assay (ELISA) for the detection of Brucella abortus antibodies in bovine bulk milk samples was evaluated. About 31 individual milk samples from B. abortus infected cows were diluted into bulk milk from a brucellosis free herd. Individual milk samples obtained from 96 negative or positive herds to ELISA or Brucella ring test (BRT), were tested by ELISA. All positive cows were bled and serum samples were tested by the complement-fixation test (CFT) which was considered the definitive test. A herd was considered infected if at least, one cow was positive in the CFT. Four samples were negative in the BRT at the dilution 1:10 but positive in the ELISA. For samples positive in both tests, BRT titers ranged from 1:10 to 1:480 while ELISA titers ranged from 1:10 to 1:3200. Using bulk milk samples, the sensitivity of the ELISA (98.1%) was higher than the BRT (72.2%) but the specificity of BRT (90.5%) was not statistically different (P=1.0) from the ELISA (88.1%). The implications of the results for brucellosis control are discussed. PMID- 11423196 TI - Sequence analysis of a RAPD band differentiating high and low virulence Staphylococcus aureus strains from rabbits. AB - RAPD typing revealed the presence of a nucleotide band in typical high virulence rabbit Staphylococcus aureus strains which was absent in low virulence strains and in an atypical high virulence strain. The nucleotide sequence of this band was determined. Primers within this sequence were developed and PCR products of eight typical high virulence, one atypical high virulence and nine low virulence rabbit S. aureus strains were sequenced. All low virulence strains and the atypical high virulence strain revealed a constant difference with the typical high virulence strains for nucleotide 377 of the 1055bp sequence. The eight typical high virulence strains possessed a guanine base on this site, while the other strains tested showed an adenine base. These findings support the hypothesis on the clonal origin of typical high virulence rabbit S. aureus strains. After comparison with databases, two open reading frames (ORF) were identified within the sequence, which appeared to encode two structural ribosomal proteins. The single nucleotide mutation does not affect the amino acid sequence of the protein it encodes for. PMID- 11423197 TI - Experimental airborne transmission of Streptococcus suis capsular type 2 in pigs. AB - Experimental airborne transmission of Streptococcus suis type 2 was studied in specific pathogen free piglets. Forty piglets were allotted to five groups of eight 7-week-old animals and housed in three separated units. Negative control pigs (group 1) were housed in unit A and infected batches were housed in units B (group 2) and C (groups 4). In units B and C, non-inoculated groups (groups 3 and 5, respectively), 40 cm distant from the respective inoculated group and without any physical contact between them, also took place. Six animals of groups 2 and 4 were inoculated intravenously with 2 x 10(8) colony forming units (cfu) of a mild and a high virulent S. suis strains, respectively. The remaining animals in these groups and pigs from groups 1, 3, 5 received broth medium in the same way. Differences among virulence of S. suis capsular type 2 were observed in inoculated pigs of groups 2 and 4. Pigs from group 2 became carriers, showing only mild symptoms. By contrast, animals from group 4 presented an acute form of the disease. All the indirect contact pigs in groups 3 and 5 had S. suis in palatine tonsils from day 6 after the infection and they presented clinical manifestations similar to those observed in experimentally infected pigs. Two direct contact animals were also contaminated in the upper respiratory tract but surprisingly they did not show any symptoms. Airborne transmission of S. suis in experimentally pigs was demonstrated in the present study. Indirect infections, as described in this study, are a more realistic way to infect pigs than other experimental procedures and may be used to further study the pathogenesis of the infection caused by this important pathogen. PMID- 11423198 TI - Virulence genes and P fimbriae PapA subunit diversity in canine and feline uropathogenic Escherichia coli. AB - In this study, a total of 118 Escherichia coli strains isolated from dogs (93) and cats (25) with urinary tract infection (UTI) were tested in a multiplex polymerase chain reaction for the presence of adhesin-encoding genes (pap, sfa, and afa), hemolysin encoding genes (hly), cytotoxic necrotizing factor 1 (cnf1) and aerobactin (aer) genes. Virulence gene frequencies detected in those isolates which had been randomly collected (68 canine strains) were: 43% pap, 57% sfa, 1% afa, 44% hly, 41% cnf1 and 34% aer. These frequencies were much higher in the remaining 50 hemolytic strains of either cat or dog origin. Virulence factor associations in the 80 hemolytic strains studied revealed that 50/80 simultaneously had two adhesin genes (pap and sfa) and two cytotoxin genes (hly and cnf1), and 15/80 in addition had the aer gene. The major structural subunit and antigenic determinant of P fimbriae of uropathogenic E. coli is PapA. Polymorphism in this subunit was studied by an F antigen-specific papA allele polymerase chain reaction in 51 canine and 22 feline pap positive E. coli strains. The most prevalent canine papA alleles were F10 (39%), F15 (37%) and F12 (35%). In feline strains F15 (50%) was more frequent, other allele frequencies were F12 (45%), F14 and F10 (27%) and F16 (23%). Only nine canine and two feline strains were negative for one of the 11 serologically defined F types of P fimbriae. Three copies of the pap operon were found in 16/51 canine and 9/22 feline UTI E. coli pap positive strains. In this study, we show that a particular combination of virulence genes appears with high frequency in dog and cat urinary tract E. coli strains (pap, sfa, hly, and cnf1). In spite of the more frequent presence of F10, F12 and F15 papA alleles in this virulence gene combination, the occurrence of different papA alleles in strains where up to three copies of the pap operon are present accounts for the observed P fimbriae diversity. PMID- 11423199 TI - Re-examination of the role of the Brucella melitensis HtrA stress response protease in virulence in pregnant goats. AB - Based on previously reported studies describing the experimental infection of pregnant goats with B. melitensis strain RWP5, we proposed that the HtrA protease plays an important role in the virulence of B. melitensis in its natural ruminant host. Subsequent studies, however, have shown that RWP5 is actually an htrA cycL double mutant. In order to definitively evaluate the role of the B. melitensis htrA in virulence, we constructed an authentic htrA mutant and examined this strain in pregnant goats. The findings of these studies indicate that the contribution of the htrA gene product to the virulence of B. melitensis in its natural host is not as great as was previously proposed. PMID- 11423200 TI - Serovars of Erysipelothrix species isolated from the tonsils of healthy cattle in Japan. AB - Serovars of 79 Erysipelothrix isolates recovered from the tonsils of healthy slaughtered cattle over a 1-year period in Japan were determined by an agar double-diffusion precipitation system using typing sera representing all the known serovars, 1 through 23 and type N, of Erysipelothrix. A total of 43 out of the 79 Erysipelothrix isolates could be classified into nine serovars but the remaining 36 isolates were untypable. Of 42 isolates identified as Erysipelothrix rhusiopathiae, 4, 6, 2, 3, 1,12, 13 and 1 isolates belonged to serovars 1b, 2, 5, 9, 12, 13, 19 and 21, respectively. One isolate belonged to Erysipelothrix tonsillarum serovar 3. PMID- 11423201 TI - A variant of the endotheliotropic herpesvirus in Asian elephants (Elephas maximus) in European zoos. AB - Newly discovered, lethal elephant endotheliotropic herpesviruses (EEHV) have been identified in both Asian (Elephas maximus) and African (Loxodonta africana) elephants. Carried by otherwise healthy African elephants they can be fatal mainly for young Asian elephants. Since zoos often harbour both elephant species, we conducted a survey on the presence of EEHV in Asian elephants from 12 European zoos, 3 circuses and 1 Israeli zoo. Here, we demonstrate that all EEHV that have affected Asian elephants so far belong to the EEHV1 group. We also describe the detection and the partial sequencing of an endotheliotropic herpesvirus variant (named EEHV1b) in Asian elephants, being either an EEHV endogenous to Asian elephants or indicating different sources (African elephants) of infection. PMID- 11423202 TI - Detection of antibodies to the nonstructural protein (NS1) of influenza A virus allows distinction between vaccinated and infected horses. AB - Antibodies to the nonstructural protein (NS1) of A/equine/Miami/1/63 (H3N8) influenza virus were detected exclusively in the sera of mice experimentally infected with A/Aichi/2/68 (H3N2) and horses infected with A/equine/Kentucky/1/81 (H3N8) or A/equine/La Plata/1/93 (H3N8), but not in those of the animals immunized with the inactivated viruses, by enzyme-linked immunosorbent assay (ELISA) using a recombinant NS1 as antigen. The results indicate that the present method is useful for serological diagnosis to distinguish horses infected with equine H3 influenza viruses from those immunized with the inactivated vaccine. PMID- 11423203 TI - Application of nucleotide sequence of RNA polymerase beta-subunit gene (rpoB) to molecular differentiation of serovars of Salmonella enterica subsp. enterica. AB - To establish a molecular differentiation method for Salmonella enterica subsp. enterica, a hyper-variable region of RNA polymerase beta-subunit (rpoB) of S. enterica subsp. enterica (I), serotype Typhimurium, and Escherichia coli were investigated through comparison of nucleotide sequence of the region. The hyper variable region was identified at 612-937 of the gene. After PCR amplification of the region in the 17 serotypes and two biotypes of serotype Gallinarum of S. enterica subsp. enterica (I), the nucleotide sequences of the region were determined and compared. All serotypes were distantly related to E. coli with 82.8-84.7% identities in nucleotide sequence while showing 96.6-100% identities with each other. According to the phylogenetic analysis based on the sequenced region with the neighbor-joining method, relatedness of biotype Gallinarum to serotype Enteritidis and biotype Pullorum was determined. Biotype Gallinarum was more closely related to serotype Enteritidis than biotype Pullorum. These results suggested that the 612-937 variable region of rpoB might be useful for molecular evolutionary analysis of serotypes of S. enterica subsp. enterica (I). PMID- 11423204 TI - An investigation into the efficacy of hatchery disinfectants against strains of Staphylococcus aureus associated with the poultry industry. AB - The efficacy of 18 commercial disinfectants was investigated using the type strain, isolate 24 (I. 24), of pulsed-field gel electrophoresis pattern related Staphylococcus aureus that have shown to be associated with clinical disease in Northern Ireland broilers. Eight quartenary ammonium compound (QAC), four peroxygen, three amphoteric (AMP), one phenolic along with two chlorine-based disinfectants were tested at their manufacturer's recommended concentration (MRC) and at three 10-fold dilutions of the MRC. The efficacy of disinfectants against I. 24 was assessed in conditions with no hatchery organic matter (HOM) and in conditions with no HOM present. In addition, 17 S. aureus strains, related and non-related to I. 24 and obtained from the poultry industry were screened for any increase in resistance relative to I. 24. All disinfectants were effective against all test strains when tested in the absence of HOM. Products from the QAC and peroxygen groups were the most potent. The performance of all disinfectants was reduced in the presence of HOM. Under these conditions all chlorine-based, two out of three AMP, and one out of eight QAC disinfectants were not effective against I. 24 when tested at the MRC. The results emphasise the importance of proper application on appropriate areas, using the correct concentration and exposure time for the disinfectant. PMID- 11423205 TI - Serotype and genotype diversity and hatchery transmission of Campylobacter jejuni in commercial poultry flocks. AB - We investigated the genotype and serotype diversity of Campylobacter coli and C. jejuni in two parent flocks of adult hens and their offspring over two rotations in order to evaluate the role of hatchery mediated transmission and/or vertical transmission of campylobacters in broiler flocks. In total, 314 C. jejuni and 32 C. coli isolates from parent and broiler flocks and from the surroundings of broiler houses were typed by flagellin gene PCR/RFLP (fla-typing), and selected isolates were also typed by serotyping and macrorestriction profiling using PFGE (MRP/PFGE). The combined typing results showed that the broiler flocks could be colonised by 1-3 different Campylobacter clones and parent flocks could be colonised by 2-6 different clones. C. coli was isolated from up to 36% of birds in one parent flock, whereas only C. jejuni was isolated from broiler flocks. C. jejuni clones from different flocks were clearly discriminated by fla-typing as well as by MRP/PFGE, except for a few cases where individual isolates belonging to two different clones were found to have altered fla-types. Similarly, one C. coli clone showed pronounced fla-type variation. The present results lead to the conclusion that vertical transmission or horizontal transmission via the hatchery are not significant transmission routes of C. jejuni to broiler chickens under Danish conditions. In the cases where more than one Campylobacter clone simultaneously colonised flocks, we found that the different clones coexisted in flocks rather than excluding each other. PMID- 11423206 TI - Improved sensitivity of PCR for Chlamydophila using pmp genes. AB - Primers targeting the conserved pmp gene family of Chlamydophila abortus were evaluated for their ability to improve the polymerase chain reaction (PCR) sensitivity. In purified DNA, specific pmp primers (named CpsiA and CpsiB) allowed at least a 10-fold increase of the PCR sensitivity compared to the specific ompA primers for C. abortus, but also for C. psittaci and C. caviae strains. No amplification was observed on C. felis, C. pecorum, C. pneumoniae and Chlamydia trachomatis strains. Tested on contaminated specimens such as genital swabs, the PCR sensitivity observed with CpsiA/CpsiB was also better than with the ompA primers. This study demonstrated that these specific pmp primers could serve as valuable, sensitive and common tools for a specific Chlamydophila diagnosis in ruminant, avian and human diseases. Digestion by AluI of the CpsiA/CpsiB fragments allowed a specific discrimination of the strains in function of their hosts and/or their serotypes. PMID- 11423207 TI - Binding of a surface protein of Staphylococcus aureus to cultured ovine mammary gland epithelial cells. AB - Staphylococcus aureus is the most persistent pathogen causing ovine mastitis. This study investigated S. aureus binding to cultured epithelial cells obtained from the mammary gland. A staphylococcal 145kDa cell wall adhesin, originally isolated from a bovine mastitis strain, was detected in lysostaphin-solubilized ovine mastitis strains and in the encapsulated strain A. This adhesin was able to bind to cultured ovine mammary gland epithelial cells (MGEC) and to a rat intestinal epithelial cell line (RIE-1), exhibiting different electrophoretic mobilities that could be attributable to protein polymorphism. Inhibition assays using antibodies against 145kDa adhesin and against whole bacteria showed the specificity of the binding to cells. The role of this protein in adherence was assessed by adherence inhibition tests carried out in vitro with radiolabeled bacteria and cultured epithelial cells. Preincubation of bacteria with antibodies against adhesin 145kDa or against strain c195 resulted in a statistically significant decrease of adherence. These experiments suggest that adherence of S. aureus to MGEC may be critical for colonization. PMID- 11423208 TI - Staphylococcus sciuri as a part of skin, nasal and oral flora in healthy dogs. AB - The coagulase-negative species Staphylococcus sciuri is widespread in nature and is associated with a variety of domestic and wild animals. However, the occurrence of S. sciuri in dogs has received little attention so far. In the present study, we established the prevalence of S. sciuri in a large population of healthy dogs, and characterized isolated strains. Samples from two mucous membrane sites (anterior nares and mouth), and two hair-coated sites (head and withers) were taken from 122 dogs and inoculated into STS agar, a novel selective medium that was introduced and tested in the study. In total, 116 isolates of S. sciuri were obtained from 488 specimens. S. sciuri was isolated from 56 out of 122 (46%) dogs. The occurrence of S. sciuri in the anterior nares and mouth were significantly higher than those in withers and head. No significant association of S. sciuri occurrence in dogs and factors such as sex, age, and living environment (indoor/outdoor) was found. Out of 56 dogs, which tested positive for S. sciuri, 30 (54%) would have it as a resident flora. Thus, we showed that S. sciuri was frequently present as a part of skin, nasal and oral flora in healthy dogs both as a resident and transient carriage. PMID- 11423209 TI - Characterization of leukocytotoxic and superantigen-like factors produced by Staphylococcus aureus isolates from milk of cows with mastitis. AB - Staphylococcus aureus is a major pathogen for cattle, causing various forms of subclinical and clinical mastitis. Two groups of virulence factors (leukotoxins and superantigens) are supposed to play an important role in the initiation and/or the exacerbation of this disease. In order to detect all known and putative members of leukotoxins and SAgs (superantigens), we tested secreted factors of different S. aureus isolates in flow cytometry-based assays. Isolates were sampled from 68 cows of different farms and cultured for 24h in vitro. Supernatants were then coincubated with purified polymorphonuclear granulocytes (PMN) or combinations of blood mononuclear cells (MNC) and PMN. Viable PMN and MNC were determined by quantitative flow cytometry. In addition, we recorded the proliferation-inducing potential of isolate supernatants for bovine MNC. Based on these criteria, the supernatants of S. aureus isolates fell in three groups. The first group (n=32), termed LT-SNs (leukotoxin-containing supernatants), killed purified granulocytes (neutrophils and eosinophils) in vitro. The second group of supernatants (n=20), termed SAg-SN (superantigen-containing supernatants), induced activation and proliferation of mononuclear cells (MNC) and, only in the presence of MNC, resulted in a selective depletion of neutrophils after 24h in vitro. The third group of supernatants (n=16) contained neither LTs or SAgs. Functionally, SAg-SNs behaved like purified staphylococcal enterotoxin A (SEA) or SEB tested in parallel. The absence of SAg-like activity in LT-SNs was confirmed by heat treatment of LT-SNs, which destroyed the leukocytotoxic activity, but did not reveal any MNC-activating potential. This study, therefore, suggests, that pathogenic S. aureus isolates either produce leukotoxins or superantigens and that both groups of virulence factors can easily be differentiated by the functional assays described. The prevalence of leukotoxin- or superantigen producing isolates was comparable among cattle with subclinical (LT=41%; SAg=30.8%) mastitis. The higher frequency of LT-producing isolates in cases of clinical mastitis (LT=55.2%; SAg=27.6%) was not significant. At least, these findings argue against the dominant role of superantigens or leukotoxins in S. aureus-induced bovine mastitis. PMID- 11423210 TI - Mathematical model of the evolution of statoconia. AB - A mathematical model of the evolution of statoconia in statocysts of freshwater snails based on the analysis of experimental data [Wiederhold et al., 1990; Pedrozo et al., 1996; Gao et al., 1997; Gao and Wiederhold, 1997; Wiederhold et al., 1999] is proposed. The growth of statoconia is considered as the process of solution crystallization. The model proposed assumes that two main processes determine the evolution of statoconia in developing snails: the generation of new statoconia and the linear growth of statoconia sizes. The analytical solution of the model and qualitative comparison of theoretical results with the experimental data show: (1) there are at least three periods of statoconia evolution; (2) the generation of new statoconia mainly determines the first period of evolution when the shell diameter of snails D < 4 mm; (3) when D > 6 mm the size distribution of statoconia is determined by the growth of their sizes with a constant rate; (4) on the interval deltaD = 4-6 mm the transformation of size distribution with selective dissolution of statoconia takes place. The model agrees well with the experimental data and makes it possible to estimate some parameters of the statoconia kinetics. Additional experiments, which are necessary for further development of the model, and quantitative estimates of the mechanisms of statoconia evolution are formulated. PMID- 11423211 TI - Metamorphic development of the bronchial columella of the larval bullfrog (Rana catesbeiana). AB - Histological and immunohistochemical analyses of head and respiratory structures in bullfrog (Rana catesbeiana) tadpoles were undertaken to address the hypothesis that the bronchial columella (BC) is the primary sound conduction pathway in these larval anurans. In postembryonic tadpoles, the BC is composed of fibroblasts surrounded by a Type I collagen matrix, with Type II collagen located in basement membranes at the distal ends. It provides a highly flexible tendon like attachment between the round window and the membranous sac of the primary bronchus of the ipsilateral lung. As the animals approach metamorphic climax stages, the fibroblasts decrease in number and the BC becomes almost exclusively collagenous. During metamorphic climax, the BC degenerates and is completely resorbed by the time the animal becomes a postmetamorphic froglet. At all larval stages examined, the BC is structurally and immunohistochemically different from both the opercularis muscle of tadpoles and the tympanic columella (stapes homolog) of postmetamorphic animals. These observations suggest that the BC may not be rigid enough to provide an effective coupling between the lungs and the round window. An alternative hypothesis for the function of the BC, based on its structure, is presented. PMID- 11423212 TI - Electrocochleogram after transection of vestibulo-cochlear nerve in a patient with a large acoustic neurinoma. AB - This study reports pre- and post-operative compound action potentials (CAPs) that were recorded from a 27-year-old woman with an acoustic neurinoma. During surgery it was necessary to totally sever her vestibulo-cochlear nerve to excise a large tumor. A pure tone audiogram changed to the scale-out pattern immediately after operation. However, CAP, the waveform of which was broadened, has been recorded 3 years post-operatively with a threshold elevation of 10 dB over the pre-operative threshold. This phenomenon suggests that CAP may originate from the extreme periphery of the auditory nerve within the cochlea. The broadening of the CAP was assumed to result from enhancement of the negative summating potential included in the CAP. We studied the effect of preceding stimulus on CAP using paired click stimuli pre- and post-operatively. A reduction of CAP amplitude in response to the second click of paired clicks was markedly suppressed in the inter-click interval between 3 and 80 ms post-operatively. We speculate that depletion of adaptation induced the abnormal CAP recovery described above and that the lateral efferent nerve system was involved in abnormal CAP adaptation with transection of the vestibulo-cochlear nerve in this case. PMID- 11423213 TI - Plasticity in the adult human central auditory system: evidence from late-onset profound unilateral deafness. AB - Experience-related changes in central nervous system (CNS) activity have been observed in the adult brain of many mammalian species, including humans. In humans, late-onset profound unilateral deafness creates an opportunity to study plasticity in the adult CNS consequent to monaural auditory deprivation. CNS activity was assessed by measuring long-latency auditory evoked potentials (AEPs) recorded from teens and adults with late-onset (post-childhood) profound unilateral deafness. Compared to monaurally stimulated normal-hearing subjects, the AEPs recorded from central electrode sites located over auditory cortical areas showed significant increases in inter-hemispheric waveform cross correlation coefficients, and in inter-hemispheric AEP peak amplitude correlations. These increases provide evidence of substantial changes from the normal pattern of asymmetrical (contralateral > ipsilateral amplitude) and asynchronous (contralateral earlier than ipsilateral) central auditory system activation in the normal-hearing population to a much more symmetrical and synchronous activation in the unilaterally deaf. These cross-sectional analyses of AEP data recorded from the unilaterally deaf also suggest that the changes in cortical activity occur gradually and continue for at least 2 years after the onset of hearing loss. Analyses of peak amplitude correlations suggest that the increased inter-hemispheric symmetry may be a consequence of changes in the generators producing the N (approximately 100 ms peak latency) potential. These experience-related changes in central auditory system activity following late onset profound unilateral deafness thus provide evidence of the presence and the time course of auditory system plasticity in the adult brain. PMID- 11423214 TI - Hearing loss associated with the modifier of deaf waddler (mdfw) locus corresponds with age-related hearing loss in 12 inbred strains of mice. AB - The modifier of deaf waddler (mdfw) and age-related hearing loss (Ahl) loci were both discovered as inbred strain polymorphisms that affect hearing loss in mice. Both loci map to the same position on chromosome (Chr) 10. The mdfw locus interacts epistatically with the deaf waddler (dfw) mutation on Chr 6, and the Ahl locus is a major contributor to AHL in several inbred strains. To investigate the possibility of allelism, we examined the correspondence of mdfw and Ahl phenotypes among 12 inbred mouse strains. The effects of strain-specific mdfw alleles on hearing loss were assessed in dfw2J/+ F1 hybrids produced from mating BALB-dfw2J/+ mice with mice from each of 12 inbred strains. F1 hybrids were then assessed for hearing by auditory-evoked brainstem response threshold analysis and classified as dfw2J/+ or +/+ by polymerase chain reaction typing. Heterozygosity for dfw2J accelerated hearing loss in F1 hybrids derived from all strains tested, except those produced with the B6.CAST + Ahl congenic strain. dfw2J/+ F1 hybrids derived from parental strains 129P1/ReJ, A/J, BUB/BnJ, C57BR/cdJ, DBA/2J, NOD/LtJ and SKH2/J exhibited a severe hearing loss by 12 weeks of age. Those derived from strains 129T2/SvEmsJ, C3H/HeJ, CBA/CaJ and NON/LtJ exhibited only a slight to intermediate hearing loss at that age. The hearing loss associated with these strain-specific mdfw alleles corresponds with previously determined Ahl allele effects, providing additional evidence that mdfw and Ahl are manifestations of the same gene. A functional relationship therefore may exist between the Ca2+ transporting activity of the dfw gene (Atp2b2) and AHL. PMID- 11423215 TI - Compound gravity receptor polarization vectors evidenced by linear vestibular evoked potentials. AB - The utricle and saccule are gravity receptor organs of the vestibular system. These receptors rely on a high-density otoconial membrane to detect linear acceleration and the position of the cranium relative to Earth's gravitational vector. The linear vestibular evoked potential (VsEP) has been shown to be an effective non-invasive functional test specifically for otoconial gravity receptors (Jones et al., 1999). Moreover, there is some evidence that the VsEP can be used to independently test utricular and saccular function (Taylor et al., 1997; Jones et al., 1998). Here we characterize compound macular polarization vectors for the utricle and saccule in hatchling chickens. Pulsed linear acceleration stimuli were presented in two axes, the dorsoventral (DV, +/- Z axis) to isolate the saccule, and the interaural (IA, +/- Y axis) to isolate the utricle. Traditional signal averaging was used to resolve responses recorded from the surface of the skull. Latency and amplitude of eighth nerve components of the linear VsEP were measured. Gravity receptor responses exhibited clear preferences for one stimulus direction in each axis. With respect to each utricular macula, lateral translation in the IA axis produced maximum ipsilateral response amplitudes with substantially greater amplitude intensity (AI) slopes than medially directed movement. Downward caudal motions in the DV axis produced substantially larger response amplitudes and AI slopes. The results show that the macula lagena does not contribute to the VsEP compound polarization vectors of the sacculus and utricle. The findings suggest further that preferred compound vectors for the utricle depend on the pars externa (i.e. lateral hair cell field) whereas for the saccule they depend on pars interna (i.e. superior hair cell fields). These data provide evidence that maculae saccule and utricle can be selectively evaluated using the linear VsEP. PMID- 11423216 TI - Comparison of the inner ear ultrastructure between teleost fishes using different channels for communication. AB - The anatomy and ultrastructure of the inner ear of three species of gouramis which differ widely in acoustic behavior were studied using scanning electron microscopy. Of the three species. Trichopsis possess a pectoral sound-producing mechanism while Macropodus and Betta lack sonic organs. The general structure of the inner ear and the shapes of the sensory epithelia are very similar, although they do differ on the posterior part of the saccular macula which is more S shaped in Trichopsis and Macropodus than in Betta. The maculae on the three species do not differ either in ciliary bundle type (cells with long kinocilia on the periphery of the maculae and cells with short kinocilia in the central region) or in hair cell orientation pattern. Quantitative measurements of hair cell densities and the size of the sensory epithelia of the saccule did not show significant differences between species. Data presented correlate with physiological results from other investigators showing similar auditory sensitivity in Trichopsis and Macropodus. The similarity in structure and function of the inner ears of gouramis on one hand, and the occurrence of sound generating organs in just one genus, suggests that hearing evolved prior to vocalization and thus acoustic communication in this taxon. PMID- 11423217 TI - A specific mitochondrial DNA deletion (mtDNA4977) is identified in a pedigree of a family with hearing loss. AB - This paper presents a family pedigree of sensorineural hearing loss in patients with a mitochondrial DNA (mtDNA) deletion. Genomic DNA screenings including myo 15 and connexin 26 were normal. MtDNA deletions are associated with many pathophysiologic conditions, including neurological disorders, sensorineural hearing loss, ischemia, cardiomyopathies and aging. Several mitochondrial disorders secondary to mutations or deletions in mtDNA have been identified in association with deafness. The present study describes a pedigree of five individuals with hearing loss who harbor a 4977 bp common aging deletion, in their mtDNA. Chromosomal analysis was normal in all affected individuals. Audiologic and molecular biologic findings of these patients suggest that the common aging deletion of mtDNA may be a predisposing factor in sensorineural hearing loss in this family. PMID- 11423218 TI - Effects of gentamicin and pH on [Ca2+]i in apical and basal outer hair cells from guinea pigs. AB - Aminoglycosides are widely used antibiotics and frequently produce acute ototoxicity. In this study we attempted to comparatively investigate the effects of gentamicin on Ca2+ influx of apical and basal outer hair cells (OHCs) isolated from guinea-pig cochlea. Since the solution of gentamicin sulfate salt is acidic (pH 3.1-3.3), we also explored the effect of external acidification on Ca2+ influx. By means of fura-2 microspectrofluorimetry, we measured the intracellular calcium concentration ([Ca2+]i) of OHCs bathed in Hanks' balanced salt solution (pH 7.40) during either a resting state or high K+-induced depolarization. Our results show that at the resting state, the baseline [Ca2+]i in apical OHCs (94+/ 2.0 nM) was slightly lower than that in basal OHCs (101.1+/-2.4 nM). By contrast, the increase in [Ca2+]i evoked by high K+ depolarization in apical OHCs was about two-fold greater than that in basal OHCs. Nifedipine (30 microM) abolished the increased [Ca2+]i in both types of OHCs, suggesting that Ca2+ influx was mainly through L-type Ca2+ channels of OHCs. While gentamicin and extracellular acidification (pH 7.14) can separately attenuate this increase in [Ca2+]i in both types of OHCs, their suppressive effects are additive in basal OHCs, but not in apical OHCs. The implications of these findings are that: (1) apical and basal OHCs behave differently in response to depolarization-increased [Ca2+]i, and (2) basal OHCs are more vulnerable to the impairment of Ca2+ entry during depolarization by a combination of gentamicin and extracellular acidification, which is correlated with the clinical observation that ototoxicity of aminoglycosides at the basal coil of OHCs is more severe than that at the apical coils. Moreover, the possibility that extracellular acidification may enhance the acute ototoxic effects of aminoglycosides should be considered especially in topical applications. PMID- 11423219 TI - Quantification of solute entry into cochlear perilymph through the round window membrane. AB - The administration of drugs to the inner ear via the round window membrane is becoming more widely used for both clinical and experimental purposes. The actual drug levels achieved in different regions of the inner ear by this method have not been established. The present study has made use of simulations of solute movements in the cochlear fluids to describe the distribution of a marker solute in the guinea pig cochlear fluid spaces. Simulation parameters were derived from experimental measurements using a marker ion, trimethylphenylammonium (TMPA). The distribution of this ion in the cochlea was monitored without volume disturbance using TMPA-selective microelectrodes sealed into the first and second turns of scala tympani (ST). TMPA was applied to perilymph by irrigation of the intact round window membrane with 2 mM solution. At the end of a 90 min application period, TMPA in the first turn, 1.4 mm from the base of ST, reached an average concentration of 330 microM (standard deviation (S.D.) 147 microM, n = 8). TMPA in the second turn, 7.5 mm from the base of ST reached a concentration of 15 microM (S.D. 33 microM, n = 5). The measured time courses of TMPA concentration change were interpreted using the Washington University Cochlear Fluids Simulator (V 1.4), a public-domain program available on the internet at http ://oto.wustl.edu/cochlea/. Simulations with parameters producing concentration time courses comparable to those measured were: (1) round window permeability: 1.9 x 10(-80 cm/s; (2) ST clearance half-time: 60 min; (3) longitudinal perilymph flow rate: 4.4 nl/min, directed from base to apex. Solute concentrations in apical regions of the cochlea were found to be determined primarily by the rate at which the solute diffuses, balanced by the rate of clearance of the solute from perilymph. Longitudinal perilymph flow was not an important factor in solute distribution unless the bony otic capsule was perforated, which rapidly caused substantial changes to solute distribution. This study demonstrates the basic processes by which substances are distributed in the cochlea and provides a foundation to understand how other applied substances will be distributed in the ear. PMID- 11423220 TI - Anemone repair proteins as a potential therapeutic agent for vertebrate hair cells: facilitated recovery of the lateral line of blind cave fish. AB - Blind cave fish use the lateral line sensory system to detect nearby objects. The fish responds to sudden perturbations in the water column by initiating startle responses in which they swim more rapidly. Normal startle responses disappear after trauma caused by a single 15 s immersion in calcium free water, but return within 5 days if the traumatized fish are treated with 'repair proteins' isolated from sea anemones. Polyclonal antibodies raised to fraction beta, a specific chromatographic fraction of repair proteins, bind to hair cells within superficial neuromasts. Likewise, biotinylated fraction beta binds to hair cells in neuromasts. Neuromast hair cells exposed to calcium free water followed by repair proteins have more compact hair bundles than do hair cells exposed only to calcium free water. We propose that anemone repair proteins replace linkages between stereocilia destroyed by exposure to calcium free water. PMID- 11423221 TI - NMDA receptor blockage protects against permanent noise-induced hearing loss but not its potentiation by carbon monoxide. AB - While a clear role has been proposed for glutamate as a putative neurotransmitter at the inner hair cell type I spiral ganglion cell synapse, the possible role of excessive glutamate release in cochlear impairment and of NMDA receptors in such a process is uncertain. The present study compares the protective effects of (+) MK-801, an NMDA receptor antagonist, and the relatively inactive isomer (-)-MK 801 against permanent noise-induced hearing loss (NIHL). The study also asks whether (+)-MK-801 can protect against the NIHL potentiation by carbon monoxide (CO). Rats (n = 6) were exposed to 100-dB, 13.6-kHz octave-band noise for 2 h after receiving injection of (+)-MK-801 hydrogen maleate (1 mg/kg), (-)-MK-801 hydrogen maleate (1 mg/kg), or saline. Other groups of animals were exposed to the combination of noise and CO (1200 ppm) after receiving (+)-MK-801 or saline. Additional subjects received (+)-MK-801, saline or CO exposure alone. Compound action potential (CAP) threshold sensitivities were compared 4 weeks after the exposures. The results show significant protection by (+)-MK-801 against the permanent CAP threshold elevation induced by noise alone, but no protective effect of (-)-MK-801. (+)-MK-801 produced limited protection against threshold shifts induced by the combination of noise and CO. Outer hair cell (OHC) loss was not protected by (+)-MK-801 administration. The data suggest that NMDA receptor stimulation may play a role in NIHL resulting from fairly mild noise exposure. The data do not support a role for NMDA receptor stimulation in the potentiation of NIHL that results from simultaneous exposure to CO and noise. PMID- 11423222 TI - Noise-induced hearing loss: the effect of melanin in the stria vascularis. AB - Conflicting investigations regarding the potential protective effect of melanin against noise-induced sensorineural hearing loss have suggested that eumelanin and pheomelanin may have differing effects within the stria vascularis. Three strains of C57BL/6J mice, (+/+, a/a) wild-types (dark coats/black eyes), (c2j/c2j, a/a), albinos (white coats/pink eyes), and (+/+, Ay/Ay) yellow mice (yellow coats/black eyes), were subjected to five consecutive days of broad band noise exposure at 112 dB(A) SPL for 3 h/day. Cochlear function was evaluated with auditory brainstem response audiometry to pure tones immediately pre-exposure, 5 6 h postexposure, and 14 days post-exposure. No significant difference in the degree of sensorineural hearing loss induced in the three strains of mice was identified. The eumelanin and pheomelanin content of each stria vascularis and amount of protein per stria for both mouse and guinea pig (2/NCR) were determined via high performance liquid chromatography. No pheomelanin was found in the stria of yellow mice, suggesting that coat color is not an accurate predictor of strial melanin content. The melanin content per mg of strial protein was higher in mice than in guinea pigs. A species-specific difference in melanin content does not explain the absence of a protective effect in mice. PMID- 11423223 TI - Quinine-induced alterations of electrically evoked otoacoustic emissions and cochlear potentials in guinea pigs. AB - Quinine is a well-known ototoxic drug which may affect portions of the auditory system with different biochemical effects, causing reversible hearing loss and tinnitus. Recent investigations indicate that quinine at high concentrations can act directly on cochlear outer hair cells to affect their motility and the mechanical response of the basilar membrane. This study aimed to investigate the effect of quinine on the electromotility of outer hair cells in vivo by means of measuring the electrically evoked otoacoustic emissions (EEOAEs), and the relationship between EEOAE and hearing sensitivity alterations in guinea pigs. Quinine was infused into the scala tympani with concentrations between 0.05 and 5 mM. An alternating current (35 microA RMS) swept from 400 Hz to 40 kHz was applied to the round window to evoke the EEOAE. The compound action potential (CAP), cochlear microphonic (CM) and summating potential (SP) were also measured. Results show that quinine affects the EEOAE in a dose-dependent manner and that its effects are reversible. Two aspects of the EEOAE were affected by quinine, depending on concentration: (1) the 'fine structure' only for concentrations below 0.1 mM and (2) the overall amplitude and the 'fine structure' for concentrations above 0.1 mM. At 5 mM the fine structure was completely absent and the mean amplitude of the EEOAE greatly decreased. Multiple component analysis shows the short delay component of the EEOAE is related to the mean value of the amplitude spectrum while the long delay component is related to the fine structure. The alterations of the EEOAE are roughly comparable to that of the cochlear potentials. A 'threshold concentration' for quinine's effects was found at 25 microM. CAP was significantly affected at 25 microM while EEOAE, CM and SP were not. Enhancement of the EEOAE amplitude was noticed in five out of 20 animals in the current study. The enhancement appears only related to the EEOAE mean level or short delay component. The results suggest that quinine can affect in vivo electromotility of outer hair cells at low concentration and therefore change the cochlear amplifier performance via an effect on electro-mechanical transduction. Its effects on the cochlear spiral ganglion neurons and/or their presynaptic process are also suggested, and these are speculated to be the primary sites for quinine's effects on the auditory system. PMID- 11423224 TI - Perilymphatic application of cisplatin over several days in albino guinea pigs: dose-dependency of electrophysiological and morphological effects. AB - Cisplatin, at 0, 3, 30 or 300 microg/ml in saline, was applied to the scala tympani of the cochlea of guinea pigs via osmotic mini-pumps, operating at a pump rate of 0.5 microl/h. Electrocochleographic recordings were made from an implanted round window electrode. When an electrocochleographic criterion of ototoxicity was reached (40 dB loss in compound action potential (CAP) threshold at 8 kHz), or after 1 week if this criterion was not reached, the animals were sacrificed for light microscopy. A subgroup of animals had endocochlear potentials (EPs) measured prior to sacrifice. Hearing remained stable in the 0 microg/ml control group, but a sudden drop of auditory sensitivity across the whole frequency range was observed in all other groups. It took 1-5 days before the drop occurred, dependent on cisplatin concentration. CAP and cochlear microphonics were lost simultaneously. The EP was severely depressed in the affected animals, suggesting that cisplatin effects on the EP are primary. However, histology revealed an accompanying loss of outer hair cells, primarily in the basal turn. It is concluded that if cisplatin is given until ototoxicity becomes apparent electrophysiologically, then the cochlear pathology from intrascalar cisplatin administration resembles that from daily parenteral administration at 1.5-2.0 mg/kg. The cochlear pathology from the parenteral treatment was greater than that observed with 30 microg/ml pumps, and less than that from 300 microg/ml pumps. PMID- 11423225 TI - Spontaneous burst-firing in three auditory cortical fields: its relation to local field potentials and its effect on inter-area cross-correlations. AB - Burst-firing refers to epochs of sharply elevated neural discharge. It has been suggested that correlated firing in different cortical areas in anesthetized animals results from spontaneous burst-firing related to electroencephalogram spindling activity and state of drowsiness. To investigate this, simultaneous recordings of spontaneous firings of neurons in the primary (AI), secondary (AII) and anterior (AAF) fields of the auditory cortex in the lightly anaesthetized cat were obtained. This allowed a study of bursting behavior in the three cortical areas under exactly the same anesthetic state. Burst occurrences were detected using the Poisson-surprise method, and were typically highly synchronized with local field potentials (LFPs) and with burst-firing of other neurons recorded on the same electrode. Burst-firing occurred in 85% of 371 units studied, and in 48 (15%) thereof there were at least 100 bursts per 15 min. Neurons in Al were bursting at a significantly higher rate, but with fewer spikes per burst, than units in AII. The average percentage of the time that a spontaneously firing neuron is in the bursting state is only about 3% (range 0.004, 29%). The average peak cross-correlation coefficients between spikes and LFP triggers were largest for burst-onset spikes, followed by those between all burst spikes and LFP triggers, and smallest when all spikes of the single unit were used in the correlation. This was the case for within- and between-area conditions. Burst onset times in different auditory fields were not correlated. Thus, the major cause of the observed correlation of spontaneous firing in different cortical areas is not synchronous burst-firing. PMID- 11423226 TI - Measurement of stapes mobility in guinea pigs and rabbits. AB - In tympanoplasty, it is essential to know the condition of the stapes. However, it has been difficult to evaluate stapes mobility in routine measurement. With the eye on improving future clinical practice, in this study we developed a new, easy system of measuring stapes mobility quantitatively and, as a first step, applied it to measurement of the relationship between the load and displacement of the stapes in guinea pigs and rabbits. The stapes displacement increased linearly with an increase in load in the small displacement region, and increased nonlinearly in the large displacement region. The slope of the regression line of this stiffness curve in the small displacement region was used as an index of the stapes mobility. The values in the guinea pigs and rabbits were 16+/-7 N/m and 115+/-25 N/m, respectively. A significant difference between the two species was observed. PMID- 11423227 TI - The Newcastle Auditory Battery (NAB). A temporal and spatial test battery for use on adult naive subjects. AB - A battery of tests for assessing the perception of temporal and spatial acoustic cues is described, together with a software platform for implementing the battery. The software runs on a personal computer either with a sound card or with widely used laboratory hardware. The battery is intended for use with neurologically impaired and other naive subjects, to allow inference at the single-subject level for any given subtest. The aim is to allow a systematic psychoacoustic evaluation of complex sound processing in single patients. Normal values are given for the threshold data for 30 naive control subjects aged from 20 to 60 years. Future modifications of the battery are allowed by modular software architecture. PMID- 11423228 TI - Styrene-induced hearing loss: a membrane insult. AB - Styrene is an aromatic solvent widely used as a precursor for polystyrene plastics in many factories which produce glass-reinforced plastic. This solvent has been shown to disrupt the auditory system in both humans and animals. In order to study the sequence of events which could explain the cochlear impairments, a time course experiment was carried out with 6-month-old rats. Male Long Evans rats were exposed to 1000 ppm styrene for 6 h/day, 5 days/week, for either 1, 2, 3, or 4 consecutive weeks. Auditory function was tested by recording the near field evoked potentials from the inferior colliculus, and histological analyses of the cochleae were performed with light and transmission electron microscopy. The electrophysiological results support a toxic mid-frequency process which keeps worsening even after the end of the exposure. The histological findings demonstrate that supporting cells are the first targets of the solvent. Then, the outer hair cells of the third row (OHC3) are disrupted, followed successively by OHC2 and OHC1 from the basal (20 kHz) to the upper turn (4 kHz) of the cochlea. Basically, the disorganization of the membranous structures could be the starting point for the cochlear injury induced by styrene. This paper presents a hypothesis that the accumulation of K+ in the spaces of Nuel underlies the toxic effects of styrene. PMID- 11423229 TI - Relationship between mammary gland infections and some milk immune parameters in Sardinian breed ewes. AB - Over the last few years an increased rate of intramammary infections caused by environmental and opportunistic pathogens has been observed in sheep farms. The presence of these microorganisms is mainly related to poor conditions of environmental hygiene and/or to decreased defenses of the mammary gland. The work we present here is a part of a project on the immune status of ewe mammary gland and its influence on mastitis development.In this paper, we have studied how intramammary infections caused by opportunistic pathogens reflect on immune parameters in ewe milk. Milk samples collected monthly before the morning milking, were screened for the presence of microorganisms and tested for somatic cell counts (SCC), polymorphonuclear neutrophil leukocytes (PMN), phagocytic activity, lysozyme content, and NAGase activity.Data showed that phagocytic activity was significantly higher in bacteriologically negative udders (156mV/1000 PMNs) than in infected udders (10mV/1000 PMNs).These results suggest that intramammary infection might be associated with a decrease of mammary gland immune status. PMID- 11423230 TI - Effect of a controlled-release urea supplementation on feed intake, digestibility, nitrogen balance and ruminal kinetics of sheep fed low quality tropical forage. AB - Four ruminally cannulated crossbred sheep (25+/-3.4kg BW) were divided into a 4x4 Latin square design to measure the effects of controlled-release urea supplement (CRUS). The basal diet consisted of 60% sugar cane tops (Saccharum officinarum), 30% full plant corn stubble (Zea mays), and 10% King grass (Pennisetum purpureum). Feed intake, digestibility, N balance and in situ ruminal kinetics were studied with four diets, D1 (control), D2, D3 and D4, which included the ratios of 100:0%, 90:10%, 80:20% or 70:30% of basal diet with CRUS. Results showed DMI differences (P<0.05) for D4 (822g per day) versus D1, D2 and D3 (580, 659 and 700g per day, respectively). N retention increased (P<0.05) for D4 (35.69g per day) versus D1, D2 and D3 (9.29, 6.85 and 19.10g per day, respectively). In vivo N digestibility was greater (P<0.05) in D4 (79.63%) than in D1 (57.57%). In vivo digestibility of DM, OM, GE, cellulose and hemicellulose was similar among the four groups. Digestibility of cell walls in D4 was higher (P<0.05) at 74.06% versus 67.78% in D1. In situ DM digestibility showed differences (P<0.05) among all diets at 9, 12, 24 and 48h of incubation. Potentially digestible fiber, 52.61%, was higher (P<0.05) in D4 versus 31.00% in D1. Indigestible fiber, 35.29%, was lowest (P<0.05) in D4 compared to 81.51% in D1. Digestion rate constant (k(d)) was different (P<0.05) between the experimental diets and control. Passage constant (k(p)) was different (P<0.05) between all diets (0.036/h in D4 to 0.081/h in D1). True digestibility was higher (P<0.05) in D4 (44.64%) compared to D1 (19.55%), but in D2 (24.54%) and D3 (28.22%) there was no difference. Cellulose in situ digestion rate, the potentially digestible fiber, was higher (P<0.05) in D3 (42.74%) as compared to D1 (22.50%). Time of disappearance of cellulose in D4 (14.79h) was less (P<0.05) than in D1 (24.03h), however there was no difference between D1 and D2. Hemicellulose in situ digestion was different (P<0.05) between D3 (45.48%) and D1 (23.61%). Digestion rate was higher (P<0.05) between D3 and D4 as compared to D1. Passage rate was different (P<0.05) between D4 (0.033/h) and D1 (0.018/h). True digestibility in D3 (34.84%) and D4 (34.62%) was higher (P<0.05) than that in D1 (20.06%) and D2 (25.86%). Half-time disappearance (t(1/2)) for hemicellulose was higher (P<0.05) in D1 (62.36h) than in D3 (28.00h) and D4 (20.64h). This study demonstrated that low quality forages at 70% of the total diet can be efficiently utilized by sheep when controlled-release urea supplementation is 30% of the feeding regime. PMID- 11423231 TI - Effects of water and feed restriction on body weight change and nitrogen balance in desert goats fed high and low quality forages. AB - The effects of water and food restriction on feed utilization, body weight changes and nitrogen balance were investigated in Sudanese desert goats given high or low quality forage. Nine male goats were employed in a 3x3 Latin square design where they were subjected to three treatments: ad libitum water and feed, ad-libitum feed/restricted water, and ad libitum water/restricted feed. These treatments were repeated using lucerne or sorghum hay. For both feed types dry matter intake (DMI) was not affected by treatments. Water intake decreased with feed restriction in goats fed sorghum hay. The ratio of water intake to DMI increased with lucerne hay compared to sorghum hay in both the control and feed restricted group. Water consumption increased with lucerne compared to sorghum hay. Body weight losses were more pronounced with water than with feed restriction but were less severe with lucerne than sorghum hay. Interactions due to treatment and feed type were significant for water intake and body weight change. With lucerne hay, except for crude protein (CP), nutrient digestibility improved with water restriction. Feeding sorghum hay, there were no effects on digestibility except for crude protein (CP) that was reduced with water restriction. Crude fiber (CF) and nitrogen free extract (NFE) digestibility increased with feed restriction in animals on sorghum hay. For all groups, CP digestibility was better with lucerne than sorghum hay. Interactions of the main effects were significant for all apparent digestibility coefficients as well as total digestible nutrients. The treatments did not have significant effects on nitrogen balance, however, nitrogen intake, retained or excreted in faeces was higher with lucerne than sorghum hay. It may be concluded that subjecting animals to water or feed restriction will have a more deleterious effect on feed utilization and nitrogen balance when using a low than a high quality forage. PMID- 11423232 TI - Softness and fatty acid composition of subcutaneous adipose tissue, and methylmalonic acid concentrations in the plasma of intensively reared lambs. AB - The aim of the present work was to study the relationships between lamb growth performance, soft adipose tissue and fatty acid composition of subcutaneous adipose tissue of intensively reared lambs, and to determine if the occurrence of soft fat, and of odd numbered (Odd FA) and methyl-branched-chain fatty acids in subcutaneous adipose tissue was related to plasma concentrations of methylmalonic acid (MMA). For this purpose, a sensitive gas chromatography assay to measure low plasma concentrations of MMA was developed and validated. In all, 49 male lambs were reared from 23.6 to 41.0kg. They were fed ad libitum with hay and pelleted concentrates containing either cereals (80%, C) or sugar beet pulp (70%, BP). Plasma concentrations of MMA were measured 12 days before slaughter. Softness score and fatty acid compositions of caudal adipose tissue were determined after slaughter. At the end of the trial, 75% of the carcasses had soft adipose tissue. The occurrence of soft fat appears to be related to high metabolisable energy intake per kg body weight. Soft fat was characterised by a higher water content, a lower proportion of saturated even-numbered fatty acids and higher proportions of Odd FA and methyl-branched-chain fatty acids in caudal adipose tissue, compared to firm fat. Despite high proportions of Odd FA and branched-chain fatty acids in caudal adipose tissue, plasma concentrations of MMA were low (2.02+/ 1.98&mgr;mol/l). No clear relationships between MMA concentrations and soft fat or the proportions of branched-chain fatty acids other than the Iso and Anteiso acids was observed. It was concluded that the present study does not support the hypothesis that the liver's capacity to metabolise MMA was exceeded. PMID- 11423233 TI - Nutritional evaluation of solar dried sardines as a ruminant protein supplement. AB - Solar dried sardines of various qualities were analyzed for nutrient content and for nutrient digestibility and nitrogen balance in sheep. Additionally, key serum enzymes and metabolites were examined to identify potential toxic effects. Dried sardine protein, ether extract and crude ash content ranged from 65 to 39%, 5.7 to 5.1%, and 22 to 51%, respectively, for high to low quality dried sardines. Visual appraisal of the dried sardines did not appear to be very reliable for determining dried sardine quality. Crude protein content was highly correlated (r(2)=0.962) with crude ash content based on the following equation (dry matter basis): crude protein%=86.0 - (0.961xcrude ash%). Therefore, crude ash could be used to estimate crude protein content and dried sardine quality. Digestibility in Omani sheep was determined on diets composed of a composite of dried sardines collected from Al-Batinah region of the Sultanate (the average crude protein content was 51%) and compared to that of a similar diet using soybean meal (also 51% crude protein) as the major protein source. The digestibility of crude protein, ether extract, total carbohydrates, digestible energy and metabolizable energy in diets utilizing dried sardines versus the soybean meal diet were 74 versus 76%, 69 versus 68%, 67 versus 69%, 64 versus 66%, and 52 versus 56%, respectively. Although, protein digestibility was similar, nitrogen balance data suggested that nitrogen in the dried sardine diet was not as efficiently utilized as nitrogen in the soybean meal diet. Key serum enzymes and metabolites did not suggest that dried sardines at 10% of the total dietary intake would cause metabolic disturbances in sheep. The study suggests that solar dried sardines may be effectively used in ruminant diets but may not have the protein by-pass value of commercial, processed fish meals. PMID- 11423234 TI - Effect of iodine level in mustard (Brassica juncea) cake-based concentrate supplement on nutrient utilisation and serum thyroid hormones of goats. AB - To assess the influence of dietary iodine (I) supplementation on nutritional performance and serum thyroid hormones of goats, 12 adult Barbari goats (average weight 18.8kg) were assigned randomly to three dietary treatments. The goats were fed a mustard (Brassica juncea) cake containing concentrate supplement along with either 0mg (control, I(0)), 0.050mg (I(50)), or 0.075mg (I(75)) I per animal per day for 180 days. Oat hay was given ad libitum as basal roughage. There was no difference in intake and digestibility of various nutrients or N retention among the three groups in a metabolism trial after 90 days of experimental feeding. A second metabolism trial conducted at 165 days post-feeding revealed that daily intake of DM, DCP and ME were 39.9, 41.1 and 44.8g, 2.64, 3.01 and 2.97g, and 366, 414 and 415kJ per unit metabolic body size, respectively, for the I(0), I(50) and I(75) groups. Retention of N by goats was 1.75, 2.58 and 2.56g per day (P>0.05) for the three groups, but one of the control animal was in negative balance. Mean live weight of I(75) animals was higher (P<0.05) at the end of the experimental feeding period with net live weight gains of 2.6, 4.8 and 5.4kg for the three groups. Mean serum concentration of triiodothyronine was 1.20, 1.23 and 1.65ngml(-1), being significantly higher in I(75) group. Mean level of thyroxine was 18.3, 24.9 and 27.4ngml(-1), significantly (P<0.05) higher in both I supplemented groups. It is concluded that supplementing I at the tested levels positively influenced the live weight gain of goats with no significant impact on the utilisation of nutrients. Serum thyroid hormones also increased in response to I supplementation. PMID- 11423235 TI - Light microscopical and ultrastructural characterization of goat preantral follicles. AB - Goat ovarian preantral follicles were morphologically and ultrastructurally described in this work. Primordial follicles are oocytes surrounded by one layer of squamous or squamous-cuboidal granulosa cells; primary follicles have a single layer of cuboidal granulosa cells, and secondary follicles are oocytes surrounded by two or more layers of cuboidal granulosa cells. At all developmental stages a thick layer of glycoproteins, the basement membrane, surrounded the preantral follicles. The quiescent oocyte is spherical or oval and it has a large eccentrically located nucleus with a conspicuous nucleolus. The organelles were uniformly distributed in the cytoplasm. A large number of vesicles were spread throughout the cytoplasm in all the oocytes. The cytoplasm of oocytes also contains numerous rounded mitochondria besides the usual organelles. As the follicle develops, the mitochondria become elongated. The communication between the oocyte and the granulosa cells is apparently mediated through endocytosis as indicated by the abundant coated pits and vesicles noted in the cortical cytoplasm of the oocyte. The oocyte plasma membrane presented projections that penetrated between adjacent granulosa cells and a few short microvilli lying parallel to the oocyte surface. In secondary follicles, patches of zona pellucida material were observed. Overall, the results indicate that the morphological and ultrastructural organization of caprine preantral follicles resembles that of other mammals. However, some particularities were observed, and that may indicate species specific differences. PMID- 11423236 TI - A short note on DNA lactotypes in the Churra and Manchega sheep breeds. AB - The structure and genetic diversity of the Spanish dairy breeds, Churra and Manchega, is analyzed by means of DNA lactotypes observed in alpha(s1)-, alpha(s2)-, beta- and kappa-ovine caseins. The inter-racial distribution of the casein genotypes is also analyzed. The two breeds differ quantitatively as well as qualitatively in their lactotypes. The proportions of unique lactotypes in Churra and Manchega reveal a high degree of within breed diversity. Breed genetic diversity is greater in Manchega (1.28) than Churra (1.53). The probability combination test indicates the existence of significant inter-racial differences. PMID- 11423237 TI - Main causes of mortalities in dairy goat kids from the Mexican plateau. AB - The causes of mortality in kids under extensive conditions in the Mexico plateau were studied. Perinatal mortality of kids (0-7 days) was lower (P<0.05) than mortality in the other age groups (8-90 days). The weights of the animals at death were extremely low, group (i) 0-7 days, 2.3kg; (ii) 7-30 days, 2.8kg and (iii) 30-90 days, 4kg. The principal finding at necropsy was white muscle disease (WMD). The WMD lesions, suggested selenium (Se) deficiency was present in 49 out of the 74 kids evaluated in this study (P<0.05). In many cases, muscular dystrophy was present with other pathological conditions. Enteric and pneumonic conditions were also important causes. In the small intestine villus atrophy was observed in 14 of the 25 kids that died with enteric problems between 8 and 30 days of age. Pneumonic lesions were present in 12 out of 74 kids. Miscellaneous conditions as starvation (five cases), hepatic conditions (three cases), omphalophlebitic infection (two cases) were also present in lower proportions. In conclusion, WMD was considered the main cause of deaths in goat kids between 8 and 90 days of age. Evaluation of Se status of soil, forages and goat tissues is recommended. PMID- 11423238 TI - Diagnosis of selenium status in grazing dairy goats on the Mexican plateau. AB - The objective of this study was to determine the concentration of Selenium (Se) in soils, forages and the tissues of kids and does under extensive grazing conditions in two regions of the Tlaxcala state, Mexico (Carrillo Puerto and Ixtenco) in the dry and rainy season. There were no differences in the concentration of Se in soil (0.051 and 0.047ppm), pH of the soil (6.1 and 5.9), concentration of Se in the forage (0.052 and 0.075ppm) and blood serum of goats (0.02 and 0.021ppm) during the dry and rainy seasons, respectively. The Carrillo Puerto region had a significantly higher content of Se in the soil (31%), soil pH (7%), content of Se in forage (25.9%), and content of Se in blood serum of goats (16%) compared to the corresponding values obtained in Ixtenco. The mean concentration of Se in the serum of kids with clinical signs of white muscle disease, was significantly lower (36.3%) compared to the kids from the same farm that were apparently healthy. The results of this study suggest that soil, forages, and goats have a marginal Se status in both regions during the dry and rainy seasons. Further research is required to find the most appropriate method to correct the Se-deficiency in goats from these regions. PMID- 11423239 TI - Some studies on nutritional muscular dystrophy in Qassim region in Saudi Arabia. Effect of administration of Vitamin E-selenium preparation to pregnant ewes on serum muscle-specific enzymes in their lambs. AB - The prophylactic effect of parentral administration of a single dose of Vitamin E selenium preparation given to pregnant ewes during the last third of pregnancy for protection against nutritional muscular dystrophy (NMD) in their lambs was studied. Based on determination of serum muscle-specific enzymes in the lambs, it was found that administration of Vitamin E-selenium preparation to pregnant ewes from a flock with a known history of NMD was associated with a significant decrease in serum muscle-specific enzymes levels in their lambs when compared with those from non-treated dams in the same flock. It was also observed that serum muscle-specific enzyme levels in lambs from non-treated dams in the flock with a known history of NMD were significantly higher than those in lambs from non-treated dams in a flock with no history of NMD (control group). The serum creatine phosphokinase (CPK) level was significantly higher in lambs of treated dams as compared to lambs from the control group, but no significant differences were seen between the two groups of lambs for the other serum muscle-specific enzymes measured. No lambs from the treated group showed clinical signs of NMD up through the time of weaning. It is concluded that administration of Vitamin E selenium preparation to pregnant ewes in a flock with a known history of NMD was effective in preventing NMD in their lambs. PMID- 11423240 TI - Standing orientation and behaviour of goats during short-haul road transportation. AB - A field study was conducted to observe the standing orientation and behaviour among a group of meat type goats during transit by road. Randomly selected goats were observed from 10 runs (10 goats per run) carried out on the same road and the duration of each run was about 50min. Two observers stood on the deck of the truck and each noted the ongoing activities of five animals. The most frequent standing orientation adopted by goats was parallel to the truck's direction of travel (24.7+/-2.1min) followed by diagonal orientation (11.9+/-1.6min) and perpendicular orientation (9.1+/-1.4min) during the 50min road journey. The goats were not observed to orient themselves opposite to the truck's direction of travel. Although, the animals changed their orientation frequently, apparently to maintain balance, there was a clear bias against the perpendicular and diagonal orientations. Mean frequencies for the various behaviours were: bleating (1.3), jumping (0.7), pushing (0.9), fighting (0.7), falling (3.1), urination (0.5), defecation (0.3) and rumination (0.1). Jumping and bleating mostly occurred at the start of the journey.Virtually all fallings were associated with the driver's changing speed of vehicle by braking (43.4%), cornering (32.4%), speed bump (12.3%) and acceleration (11.9%). These falls, which hamper the well being of animals in transit and are also responsible for bruising which can be prevented by careful driving and by adjusting the speed of the vehicle gradually. PMID- 11423241 TI - Editorial. PMID- 11423242 TI - Kidney ultrasound and Doppler ultrasound findings during and after acute urinary retention. AB - OBJECTIVE: To evaluate changes in kidney ultrasound and Doppler ultrasound images during and subsequent to acute urinary retention (AUR). METHODS: Twenty-five men with a mean age of 69 years suffering AUR for a mean of 31 h were studied by measuring serum creatinine, creatinine clearance and renal ultrasound. Renal Doppler ultrasound was applied in 19 of these cases and all patients were followed for 6 months after acute retention was relieved. RESULTS: During AUR hydronephrosis was noted in three patients; this disappeared during follow-up. During the acute period, after 1 month and after 6 months the average resistive indexes (RI) were 0.71, 0.70 and 0.69, respectively. The changes were not statistically significant. During follow-up, the proportion of patients with normal RI increased from 42 to 64%. Median serum creatinine was normal during retention and follow-up. Median creatinine clearance was reduced during retention and became normal during follow-up (P < 0.05). No correlation was found between RI and serum creatinine at any time-point, nor was any correlation noted between RI and creatinine clearance during retention or at the 1-month follow-up; at 6 months, however, there was a significant inverse correlation between them (P = 0.01). CONCLUSION: AUR caused elevation of RI, which may be interpreted as diminished renal blood flow. Although in the majority of patients it recovered after treatment, elevated RI was still found in one third of the patients, possibly due to previous chronic bladder outlet obstruction. Our findings stress the importance of both fast release of AUR and effective treatment of its cause. PMID- 11423243 TI - Comparison of power Doppler and velocimetry in predicting outcome of high-risk pregnancy. AB - OBJECTIVE: Prospectively evaluate semi-quantitative computer analysis of power Doppler (PD) signals in the placenta, tetal brain, lung, liver, kidney and spleen in high-risk pregnancies in relationship to perinatal outcome and also to compare tissue blood flow in the fetal brain and placenta with Doppler velocimetry. METHODS: PD signals were recorded in 180 high-risk pregnancies between 27 and 41 weeks of gestation. Images from PD angiographic scans were transmitted for computer analysis of pixel intensity. Mean flow signal intensity was recorded for each organ. The PD brain/lung ratio was calculated. The PD results were plotted on reference values and related to perinatal outcome. Middle cerebral (MCA), umbilical (UA) and uterine artery (Ut. A) velocimetry was also performed. RESULTS: High-risk pregnancies displayed lower PD signal intensity from the placenta, fetal lung, liver and kidney as compared to normals. However, the brain and spleen signals showed higher intensities suggesting increased tissue perfusion. PD signals from the fetal brain, lung, placenta and PD brain/lung ratio were correlated with perinatal outcome. The PD signal intensity from the fetal liver, kidney and spleen showed poor correlation with perinatal outcome. Fetal brain tissue blood flow showed better correlation with the outcome than MCA velocimetry. Placental tissue blood flow results were similar in predicting outcome to those obtained by means of UA and Ut. A velocimetry. CONCLUSION: In comparison with conventional Doppler velocimetry, computer analysis of PD signals, give similar results in the prediction of adverse perinatal outcome. PMID- 11423244 TI - Cerebral blood flow velocity and inflammatory response after severe traumatic brain injury. AB - OBJECTIVES: The cerebral blood flow velocity (CBVF) was measured by transcranial Doppler sonography in patients with severe traumatic brain injury (TBI) in order to determine, whether it depends on the posttraumatic inflammatory response. MATERIAL AND METHODS: CBVF in both middle cerebral arteries (MCA) was recorded in 25 comatous TBI patients (male 20; female five; mean age +/- standard deviation (S.D.), 41 +/- 20 years) and correlated to the levels of interleukine-(IL) 6, IL 8 and IL-10 in corresponding CSF/plasma samples, to PaCO2 and to intracranial (ICP), mean arterial (MAP) and cranial perfusion pressure (CPP). RESULTS: CSF IL 6 and IL-8 were clearly higher than the corresponding plasma levels (mean CSF/plasma quotient for IL-6: 159 +/- 582; for IL-8: 143 +/- 311). CBVF did not show large side-to-side differences at each examination indicating that CBFV in both MCAs was determined mostly by systemic conditions and not by severe regional abnormalities. Since all other evaluated variables including interleukines represent also systemic conditions we used the mean value (MCBFV) of both CBFVs for analysis. By stepwise regression analysis between MCBVF (mean +/- S.D., 80 +/ 26 cm/s) and the variables PaCO2 (33 +/- 4 mmHg), MAP (86 +/- 12 mmHg), ICP (20 +/- 11 mmHg), CPP (70 +/- 14 mmHg) and CSF or plasma IL-6, IL-8 and IL-10, it turned out that MCBFV correlated significantly with PaCO2 (r = 0.478; P < 0.01) and CSF IL-8 (r = -0.361; P < 0.05). CONCLUSIONS: When CPP is adequate for brain perfusion, CBFV in large brain supplying arteries depends predominantly on PaCO2 and shows only a slight association to intrathecal IL-8 levels. For clinical interpretation of CBFV data, the inflammatory response seems to be of minor relevance. PMID- 11423245 TI - Coordinated ultrasound screening of infants: Hungary experience. AB - OBJECTIVE: In 1990, three hospitals (the Madarasz Street Children's Hospital in Budapest and the general hospitals in Cegled and Zalaegerszeg) organised a coordinated ultrasound (US) screening programme. The screening was aimed at detecting a predefined series of developmental and acquired disorders. MATERIAL: In 9 years, a total of 46,858 infants were screened by the three hospitals: 20,185 in Budapest, 13,258 in Cegled and 12,415 in Zalaegerszeg. RESULTS: Disorders were detected in 4068 cases. Most (2885) involved abdominal or renal disorders; intracranial anomalies were found in 1062, and the remaining discoveries were of cysts or tumours in the liver, spleen or ovaries. CONCLUSIONS: Many disorders were detected (8.7%) needing either follow-up, or some form of treatment. PMID- 11423246 TI - Endosonography for assessment of anorectal changes in patients with fecal incontinence. AB - Our aim was to evaluate the anorectal changes in patients with fecal incontinence by using different endoultrasound techniques. Eighteen females with fecal incontinence and 14 healthy controls (females) were examined. Rigid linear transducer and 10 MHz radial scanning miniprobe were used. Statistically significant difference was found in the mean percentage of decreasing of the anorectal angle during contraction between healthy subjects (21.4%) and incontinent patients (7.5%). The mean percentage of shortening of the puborectal muscle length during squeezing was significantly lower in patients (9.6%) than in controls (17.9%). By means of miniprobe the anal sphincter defects were clearly demonstrated. Endosonography with a radial scanning miniprobe and rigid linear transducer ensures complex morphological and functional assessment of the anorectal region. PMID- 11423247 TI - Uncommon ultrasound findings in traumatic extracranial vertebral artery dissection. AB - We report a case of internal carotid artery dissection (ICAD) associated with contralateral vertebral artery dissection (VAD). The interest of this case is to discuss an unusual Doppler pattern manifesting by a spectrum of an alternating vertebral artery flow suggesting a hemodynamic contribution from the contralateral vertebral artery (VA) and a clear depiction of both antegrade (red) and retrograde (blue) flow within the false and true lumen of the VAD by color Duplex flow imaging. PMID- 11423248 TI - A review of two alternative ultrasound quality assurance programmes. AB - OBJECTIVE: This study reviewed the results of B-mode Quality Assurance (QA) performance tests on 17 real-time ultrasound scanners, performed over a period of 3 years, in order to assess their value. Following this review we revised and simplified our testing schedules to include two tests for noise and sensitivity. The value of the new schedules was assessed. METHODS: Initially, testing schedules were similar to those recommended by two professional bodies. Results were reviewed to determine whether the tests predicted or confirmed faults. We then introduced a simplified testing programme using alternative measurements, attempting to demonstrate or predict noise related faults that affect the image, but were not demonstrated by current tests. These new tests have been performed on 24 ultrasound machines for up to 18 months. RESULTS: A review of results has shown that measurements occasionally fall outside tolerance due to chance, and that faults that significantly affect the image, e.g. probe faults and noise, are reported by the users without predictive or concomitant changes in test results using our original schedules. Faults occur that do not immediately affect image quality and are not reported by the users. Inappropriate settings, e.g. monitors, are frequently reset at QA, particularly where there are potentially untrained users. The additional tests showed consistent changes in noise (four) or sensitivity (one) on five machines. CONCLUSION: Our earlier tests were inadequate in demonstrating deterioration in the clinical performance of ultrasound imaging equipment. Introduction of a revised testing schedule has resulted in changes in equipment performance being detected and rectified. PMID- 11423250 TI - Acquired immunodeficiency syndrome-related cancer. A study model for the mechanisms contributing to the genesis of cancer. AB - The study of AIDS and cancer has resulted in an important convergence of clinical epidemiological investigations, molecular research and virological studies. This shared effort has led to significant discoveries in the field of human carcinogenesis. In a relatively short time, several topics have been clarified, namely: (1) the spectrum of AIDS-related tumours has been identified; (2) the role of HIV in lymphomagenesis and in the development of Kaposi's sarcoma (KS) has been defined; (3) AIDS-related cancers have allowed the discovery and extensive biological characterisation of the novel virus human herpes virus 8 (HHV8), also called KS-associated herpesvirus; and (4) genetic, molecular, and phenotypic studies of AIDS-related lymphomas have contributed to the formulation of a pathogenetic and histogenetic model of B-cell non-Hodgkin's lymphomas (NHL). PMID- 11423251 TI - Epidemiology of AIDS-related tumours in developed and developing countries. AB - AIDS-associated illnesses include Kaposi's sarcoma (KS), non-Hodgkin's lymphoma (NHL), and, since 1993, invasive cervical cancer (ICC). Between 1988 and 1998, among AIDS cases reported in western Europe, 9.6% had KS and 3.9% had NHL as AIDS defining illnesses. Between 1988 and 1998, the frequency of KS decreased from 13.4 to 6.4%, while NHL increased from 3.8 to 5.3%. Estimates of the relative risk (RR) of neoplasms in HIV-seropositive populations came from population-based cancer and AIDS registries linkage studies conducted in the United States, Italy and Australia and from a few cohort and case-control studies. In adults with HIV/AIDS, the RR was over 1000 for KS and ranged between 14 for low-grade NHL and over 300 for high-grade NHL. For Hodgkin's disease (HD), a consistent 10-fold higher RR was observed. For cervical and other anogenital tumours associated with human papilloma virus, risk increases were 2- and 12-fold, depending upon location. In Africa, the AIDS epidemic led to KS becoming the most common cancer type in men in several areas. The RR of AIDS-associated tumours were lower in Africa than those reported in western countries. PMID- 11423252 TI - Cancer processes in immunodeficient populations: an introduction. PMID- 11423253 TI - Epstein-Barr virus associated lymphoproliferations in the AIDS setting. AB - Epstein-Barr virus (EBV) is a ubiquitous gammaherpesvirus that is associated with a variety of malignancies. In vivo infection of B lymphocytes is initially associated with the broad expression of immunodominant viral latency genes and proliferation of infected cells. Ultimately, a viral reservoir is established in resting B cells with restricted expression of viral latency genes and no expression of immunodominant viral genes. Among the tumours associated with EBV that are relevant to a consideration of EBV in HIV-associated malignancies are posttransplant lymphoproliferative disease, Burkitt's lymphoma (BL) and Hodgkin's disease (HD). BL carries whereas EBV in only a minority of cases whereas HD in patients infected with HIV is virtually always EBV-associated. EBV-directed T cell therapies have proven effective in posttransplant lymphomas in bone marrow transplantation patients. In patients with HIV infection, primary central nervous system (CNS) and immunoblastic lymphomas show similarities with posttransplant lymphoproliferative disease. EBV detection studies in cerebrospinal fluid are useful diagnostically in primary CNS lymphoma. T cell therapies may be useful in the treatment of EBV-associated lymphomas. Thus, a better understanding of the relationship between EBV and these tumours will not only help to clarify their pathogenesis, but may facilitate the development of new diagnostic and therapeutic strategies. PMID- 11423254 TI - KSHV/HHV8-associated lymphoproliferations in the AIDS setting. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) or human herpesvirus 8 (HHV8) is associated with two lymphoproliferative disorders in the AIDS setting, primary effusion lymphoma (PEL) and the plasma cell variant of multicentric Castleman's disease (MCD). In PEL, KSHV persists in a latent form in most lymphoma cells, although viral production has been seen infrequently. In MCD, the viral gene expression pattern is less restrictive, virus production appears to occur and to correlate with the severity of this disease. Several viral genes may contribute to the particular features of these two disorders: among them a viral homologue of interleukin 6 (vIL6) has attracted much attention and been shown to promote the growth of plasma cells. It is thought that its activity is important in the pathogenesis of both PEL and MCD. Other viral genes, in particular a D-type cyclin homologue, the latent nuclear antigen LANA, and one or more of the viral homologues of interferon regulatory factors (vIRFs) may also contribute. Although it is conceivable that viral infection per se could explain much, if not all, of the features of MCD, it is likely that additional genetic alterations play a role in the pathogenesis of PEL. PMID- 11423255 TI - HPV-related neoplasias in HIV-infected individuals. AB - Human papillomavirus (HPV) infection of the lower genital tract is now considered the most important factor in the initiation of neoplasia. Human immunodeficiency virus (HIV) infection appears to alter the natural history of HPV-associated oncogenesis, but its impact on gynaecology has only recently been defined; the Centers for Disease Control (CDC) designated moderate and severe cervical dysplasia as a category B defining condition, and invasive cervical cancer as a category C defining condition of AIDS in 1993. Anal HPV infection and anal squamous intra-epithelial lesions have been found to be highly prevalent among HIV-positive homosexual men, and recent preliminary data suggest a relatively high prevalence among HIV-positive women as well. Moreover, HPV infection and associated lesions are also observed in body sites other than the anogenital area, particularly the skin and the oral cavity. PMID- 11423256 TI - Pathology of AIDS-related lymphomas and other AIDS-defining neoplasms. PMID- 11423257 TI - Biology of Kaposi's sarcoma. AB - Kaposi's sarcoma (KS) is an angioproliferative disease occurring in several different clinical-epidemiological forms that, however, share the same histological traits and are all associated with infection by the human herpesvirus 8 (HHV8). KS initiates in a context of immune dysregulation characterised by CD8+ T cell activation and the production of Th1-type cytokines that induce a generalised activation of endothelial cells leading to adhesion and tissue extravasation of lympho-monocytes, spindle cell formation and angiogenesis. These phenomena are triggered or enhanced by infection with HHV8 that, in turn, is reactivated by the same cytokines. Productively-infected circulating cells are recruited into 'activated' tissue sites where HHV8 finds an optimal environment for establishing a persistent, latent infection of KS spindle cells (KSC). HHV8 dissemination is favoured by virus escape mechanisms and immune dysregulation, and leads to immune responses that are not effective against the virus but, paradoxically, exacerbates the reactive process. Although early KS is a reactive process of polyclonal nature that can regress, in time it can progress in to a true sarcoma. The progression of KS appears to be due to the deregulated expression of oncogenes and oncosuppressor genes, to the long-lasting expression of the HHV8 latency genes and, for AIDS-KS, is promoted by the proliferative and angiogenic effects of the HIV-1 Tat protein. PMID- 11423258 TI - Genetic pathways and histogenetic models of AIDS-related lymphomas. AB - Acquired immunodeficiency syndrome (AIDS)-related lymphomas consistently display a B-cell phenotype and are histogenetically related to germinal centre or post germinal centre B cells in the overwhelming majority of cases. The pathogenesis of AIDS-related lymphoma is a multistep process involving factors provided by the host as well as alterations intrinsic to the tumour clone. The molecular pathways of viral infection and lesions of cancer-related genes associated with AIDS related lymphomas vary substantially in different clinicopathological categories of the disease and highlight the marked degree of biological heterogeneity of these lymphomas. PMID- 11423259 TI - Pathogenetic and histogenetic features of HIV-associated Hodgkin's disease. AB - Compared with the cases in the general population, Hodgkin's disease (HD) arising in the HIV setting shows distinctive features in terms of epidemiology, aetiopathogenesis, histopathology and clinical behaviour. Although HD does not represent an AIDS-defining condition, recent evidence consistently indicates that HIV-infected individuals have a significantly increased risk of developing HD. HIV-related HD is characterised by the preponderance of aggressive histological subtypes, advanced stage at presentation, and highly malignant clinical course. Moreover, unlike HD in the general population, the large majority of HIV-related HD cases are pathogenetically linked to Epstein-Barr virus (EBV), with rates of EBV positivity ranging from 80 to 100%. Hodgkin and Reed-Sternberg cells of these cases invariably show a strong expression of the EBV-encoded latent membrane protein-1 (LMP-1), which functions as a constitutively activated tumour necrosis factor (TNF) receptor-like molecule. Usurpation of physiologically relevant pathways by LMP-1 may lead to the simultaneous or sequential activation of signalling pathways involved in the promotion of cell activation, growth, and survival, contributing thus to most of the features of HIV-related HD. PMID- 11423260 TI - Clinical aspects and management of AIDS-related Kaposi's sarcoma. AB - AIDS-related Kaposi's sarcoma (KS) is a tumour of vascular endothelium, which is seen predominantly in men who have sex with men. The majority of affected individuals have advanced immunosuppression at the time of the initial KS diagnosis. The disease may present with cutaneous lesions, or with involvement of visceral organs, of which the gastrointestinal tract is most common. KS may also present with lymphoadenopathy or with isolated lymphoedema, even in the absence of cutaneous lesions. Affected individuals are uniformly co-infected with HIV and with Human Herpesvirus type 8 (HHV8). HHV8 is present within KS tissues, and is aetiological in the pathogenesis of disease, along with aberrant cytokine expression, production of multiple angiogenic peptides, and immune dysregulation. While not presently curable, multiple treatment options exist and must be evaluated in terms of the specific needs of the individual patient. Various local therapies are aimed at eradicating small lesions, while acknowledging that the KS in general, or its likelihood of recurring will be unaffected. Systemic chemotherapy is used to treat extensive visceral involvement. Knowledge of the pathogenesis of disease has led to the development of novel treatment strategies, aimed at HHV8 as the target of therapy, or at the inflammatory cytokine or angiogenic milieu necessary for KS growth. Use of highly active anti-retroviral therapy, aimed at controlling the underlying HIV infection, has been associated with a dramatic decrease in the incidence of KS, and may also be useful in the treatment of existing KS disease. PMID- 11423261 TI - Clinical aspects and management of AIDS-related lymphoma. AB - The incidence of non-Hodgkin's lymphoma (NHL) is increased by approximately 100 fold in patients with advanced HIV infection. Clinical presentations may include systemic lymphoma, primary central nervous system (CNS) lymphoma, and primary effusion lymphoma. Systemic lymphoma is the most common presentation, is almost always of intermediate or high-grade histology and B-cell phenotype, and usually involves extranodal sites. The disease is potentially curable with combination chemotherapy used for immunocompetent patients with lymphoma, although cure is achieved in only approximately 10-35% of patients. Primary CNS lymphoma may be difficult to distinguish from cerebral infection. The prognosis is very poor, although approximately 10% of patients selected for therapy may survive beyond 1 year with brain irradiation. Attention to infection prophylaxis and antiretroviral therapy is important. Evidence suggests that highly active antiretroviral therapy (HAART) has resulted in a decreased incidence of lymphoma, and that patients with systemic lymphoma treated in the post-HAART era have a better prognosis. PMID- 11423262 TI - Clinical aspects and management of Hodgkin's disease and other tumours in HIV infected individuals. AB - As the AIDS epidemic advances, the spectrum of malignancies encountered is expanding. Several non-AIDS defining cancers, i.e. Hodgkin's disease (HD), anal and testicular cancer, are increasing in incidence in HIV-infected patients. The widespread use of highly active antiretroviral therapy (HAART) in industrialised countries has resulted in substantial improvement in the survival of HIV-infected patients. It is likely that in the future, cancers associated with long-term mild immune suppression will occur at an increased rate in long-term survivors of HIV infection. The natural history of the majority of non-AIDS defining tumours differs from that of the general population. Unusual aspects of tumour localisation, growth behaviour and therapeutical responses distinguish tumours in patients with HIV infection from those without. This paper reviews the most relevant data on the epidemiology, pathology, clinical features and treatment of the most frequently reported non-AIDS defining tumours, i.e. HD, lung, testicular and skin cancers. PMID- 11423263 TI - AIDS and cancer in the era of highly active antiretroviral therapy (HAART). AB - Combination therapy with protease inhibitors and nucleoside analogues dramatically suppresses plasma HIV-1 RNA and delays progression to AIDS, but the impact on HIV-associated malignancy remains to be established. Observational and time-trend data indicate that the incidence of Kaposi's sarcoma (KS) and primary brain lymphoma have decreased, but suggest that current therapies have not had a proportionate effect on systemic non-Hodgkin's lymphomas (NHL). As opportunistic infection and mortality are yielding to advances in antiretroviral therapy, lymphoma may increase in importance as a cause of AIDS-related morbidity and mortality. Further improvements in the long-term consequences of HIV infection will depend on better prevention and treatment of this serious malignant complication. PMID- 11423264 TI - Impact of HAART on the clinical management of AIDS-related cancers. AB - The development of HIV-related disease has changed dramatically since the introduction of highly active antiretroviral therapy (HAART) into clinical practice. Since the use of protease inhibitors became widespread, a 30-50% reduction in Kaposi's sarcoma (KS) has been observed. The results of recent studies indicate that HAART may be a useful alternative both to immune response modifiers during less aggressive stages of KS disease and to systemic cytotoxic drugs in the long-term maintenance therapy of advanced KS. The impact of HAART regimens on the incidence of systemic lymphoma (NHL-HIV) remains unclear, but it can be hypothesised that patients treated with HAART may survive longer with continued B cell stimulation and dysregulation resulting in an increased incidence of lymphoma over time. The impact of HAART on survival in patients affected by NHL-HIV has recently been evaluated and a positive correlation between HAART and outcome in these patients has been found. The spectrum of cancers in patients with HIV infection may develop further since these patients survive longer with HAART and with a better control of opportunistic infections. With the increasing use of HAART, the dilemma is whether to institute or continue protease inhibitors use during chemotherapy. Based on the advances in our understanding of HIV-related disease and the availability of new antiretroviral therapies, the choice for anti-HIV agents in patients receiving chemotherapy is important. PMID- 11423265 TI - Is it necessary to resect the diseased esophagus in performing reconstruction for corrosive esophageal stricture? AB - OBJECTIVE: The incidence of carcinoma of the esophagus among patients with chronic esophageal stricture caused by ingestion of corrosive agents is reported to be significantly higher than that of the general population. The question of whether or not a resection of the diseased esophagus should be included in the surgical reconstruction procedure of the undilatable esophageal stricture continues to be a controversial. METHODS: During the 12 year period from 1988 to 1999, a total of 54 consecutive patients with caustic stricture of the esophagus were treated in our department. We retrospectively reviewed these cases and analyzed the incidence of cicatrical carcinoma among the patients and the risk of esophagectomy according to the procedures performed. RESULTS: We found seven cases of esophageal cancer among these patients. There was no significant increase in mortality or morbidity related to esophagectomy. CONCLUSIONS: Considering the high incidence of cicatrical carcinoma from the stricture sites as well as the possible chance of hidden malignancy, we concluded that the simultaneous resection of the esophagus with reconstruction for patients with chronic intractable caustic stricture would give the patients a better probability of being completely cured of the disease. PMID- 11423266 TI - Esophageal perforation: life threatening complication of endotracheal intubation. AB - OBJECTIVE: To raise awareness of this complication of tracheal intubation, to emphasize the gravity due to delayed diagnosis, and to advocate a surgical treatment. METHODS: Between April 1980 and January 2000, 97 patients were treated for esophageal perforation in our department. We reviewed the cases of perforation occurring after attempted tracheal intubation. Each case is presented. Discussion is focused on diagnosis and treatment. RESULTS: Esophageal perforation occurred after attempted endotracheal intubation in five cases among 58 iatrogenic perforations. There were four women and one man (mean age 72 years). In all cases, it was for a planned operation. Intubation was performed by a single lumen tube in three cases and a double lumen tube in two cases. Presenting symptoms were acute in one case and insidious in four cases. Free interval before diagnosis and treatment was long in all but one case, with an average of 179 h (range 5--432). Two patients suffered from septic shock when they were transferred. All patients were operated on. Two patients died. CONCLUSION: Post intubation esophageal perforation is one of the most life threatening esophageal perforation. Delayed diagnosis is the first cause of gravity. Prevention of this complication begins with recognition of a potentially difficult intubation. Good outcome follows from rapid diagnosis and early surgical treatment. PMID- 11423267 TI - Primary tracheal tumors: experience with 14 resected patients. AB - OBJECTIVE: Primary tracheal tumors are rare. Management includes interventional endoscopy, surgery and radiotherapy. METHODS: Between 1987 and 1996, 14 patients treated by resection and reconstruction of the trachea and bifurcation for primary tracheal tumors were retrospectively analyzed. RESULTS: The most common histological finding was adenoid cystic carcinoma (n=7), followed by a squamous cell carcinoma (n=2), a mucoepidermoid carcinoma (n=2), a carcinoid tumor (n=1) and two benign tumors (xanthogranuloma, pleomorphic adenoma). Various reconstruction techniques were used and one prosthesis was implanted. Eight of the patients required preoperative Nd-YAG laser recanalisation. Six were treated by postoperative external beam radiotherapy, in three cases combined with endoluminal brachytherapy. Two major postoperative wound-healing impairment at the anastomosis occurred. Four minor wound-healing disorders were successfully treated by interventional endoscopy. Two patients died postoperatively with mediastinitis respectively with bilateral pneumonia. A local recurrence was observed in only two cases. At the last follow-up in January 1998, nine patients were still alive. We observed five long-term survivors (>6 years) with an adenoid cystic carcinoma or mucoepidermoid carcinoma. CONCLUSIONS: Extensive segmental resection of the trachea is the treatment of choice for primary malignant and occasionally for benign tracheal tumors. Interventional endoscopy is a part of modern tracheal surgery. PMID- 11423268 TI - Surgery for bronchiectasis. AB - OBJECTIVE: The incidence of bronchiectasis has declined markedly in developed countries. However, a reasonable number of patients still need surgery, despite aggressive physiotherapy and antibiotic therapy. We have reviewed our patients to clarify the benefits from surgery and to analyse the complications. MATERIAL AND METHODS: Between 1988 and 1999, we have operated on 119 patients with bronchiectasis, 71 female and 48 male, with a mean age of 42.2 years (range 11- 77 years). Surgery was indicated because of unsuccessful medical therapy in 66 patients (55%), 31 (26%) had haemoptysis, 11 (9.2%) had lung abscess, 10 (8.4%) had lung masses, and three (2.5%) had pneumothorax. The most common manifestations were cough with sputum in 90 patients (76%), haemoptysis in 45 (38%) and recurrent infections in 57 (48%). The mean duration of the symptoms was 4 years (range 1--40 years). The lower lobes were diseased in 61 patients and bilateral disease was found in ten. The mean number of involved pulmonary segments was five (range 1-15). A lobectomy was performed in 75 patients (62%), a segmentectomy in 12 (10%), a pneumonectomy in nine (7.4%) and a bilobectomy in four (3.3%). Complete resection of the disease was achieved in 108 cases (91%). RESULTS: There was no operative mortality and perioperative morbidity occurred in 15 patients (15%), including temporary broncho-pleural fistulae in 7 (5.8%), and post-operative haemorrhage and atrial arrhythmias in four (3.3%) each. After a mean follow-up was 4.5 years, 73 patients (68%) of this group were asymptomatic, and 31 (29%) had meaningful clinical improvement, while only four (3.7%) maintained or worsened prior symptoms. The best clinical improvement occurred in patients with complete resection of the disease (P=0.008). There were no differences in the respiratory function, comparing pre- and post-operative data, with a 2-year of minimum interval. The VC was 91 and 89% and the FEV1 was 83% and 81% of expected, respectively before and after surgery, (P=NS). CONCLUSION: Surgery of pulmonary bronchiectasis has few complications and markedly improves symptoms in the great majority of patients, especially when complete resection of the disease is achieved. Pulmonary resection of bronchiectasis does not alter respiratory function. PMID- 11423269 TI - Role of physiological lung exclusion in difficult lung resections for massive hemoptysis and other problems. AB - OBJECTIVES: Pulmonary tuberculosis and bronchiectasis are the major causes of massive hemoptysis in developing countries. Lung resection remains the surgical treatment of choice. This may not always be possible and may even be hazardous in some patients due to fibrosis and dense vascular adhesions between the lung and the chest wall. This leads to marked blood loss and control of hilar vessels becomes dangerous. METHODS: A series of 20 cases is described here. Nineteen presented with massive hemoptysis where control of bleeding was obtained by physiological lung exclusion. One patient had traumatic left main bronchus transection not suitable for repair or resection. Physiological lung exclusion was performed by surgical interruption of the bronchus and pulmonary artery of the involved lobe or lung, keeping pulmonary veins intact. RESULTS: Hemoptysis could be controlled in all these patients without any significant morbidity. There was no mortality. There was no postoperative empyema and recurrence of hemoptysis on long-term follow-up. No patient required anatomical lung resection later on. CONCLUSIONS: Physiological lung exclusion is a safe and effective method for control of massive hemoptysis in cases where lung resection is technically hazardous or difficult. This should be kept as an alternative or adjunct to anatomical lung resection. PMID- 11423270 TI - Frequency and mortality of acute lung injury and acute respiratory distress syndrome after pulmonary resection for bronchogenic carcinoma. AB - OBJECTIVE: We reviewed the frequency and mortality of acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) in our population of patients submitted to pulmonary resection for primary bronchogenic carcinoma. METHODS: From January 1993 to December 1999, a total of 1221 patients received pulmonary resection for primary bronchogenic carcinoma. Of these, 27 met the criteria of post-operative ALI/ARDS. There were 24 men and three women with a mean age of 64 years (range 45--79). Pre-operatively, predicted mean of PaO(2), PaCO(2) and %FEV1 were 72 mmHg (57--86), 37 mmHg (33--42) and 80% (37--114), respectively. Associated cardiac risk factors were present in eight patients. Three patients (11%) had pre-operative radiotherapy. Surgical-pathologic staging included 14 patients at Stage I, 8 patients at Stage II, four patients at Stage IIIa and one patient at Stage IIIb. RESULTS: ALI/ARDS occurred in 2.2% of our operated lung cancer patients. ALI was diagnosed in 10 patients and ARDS in 17 patients. The mean time of presentation following surgery was 4 days (range 1--10) and 6 days (1--13) for ALI and ARDS, respectively. According to the type of operation, the frequency was highest following right pneumonectomy (4.5%), followed by sublobar resection (3.2%), left pneumonectomy (3%), bilobectomy (2.4%), and lobectomy (2%). The frequency following extended operations was 4%. No differences were found between the ALI/ARDS group and the total population of resected lung cancer patients (control group) with respect to sex, mean age, pre-operative blood gases, %FEV1, surgical--pathologic staging and the use of pre-operative radiotherapy. Four patients with ALI (40%) and 10 patients with ARDS (59%) died. Mortality was highest following right pneumonectomy, extended operations and sublobar resections. Hospital mortality of the total population of operated lung cancer patients in the same period was 2.8% (34 patients). ALI/ARDS accounted for 41% of our hospital mortality. CONCLUSIONS: (1) ALI/ARDS is a severe complication following resection for primary bronchogenic carcinoma. (2) We did not detect any significant difference between the ALI/ARDS group and the control group regarding age, pre-operative lung function, staging and pre-operative radiotherapy. (3) ALI/ARDS is associated with high mortality, the highest mortality rates having been observed following right pneumonectomy and extended operation; it currently represents our leading cause of death following pulmonary resection for lung carcinoma. (4) ALI/ARDS may also occur after sublobar resections with an associated high mortality rate. PMID- 11423271 TI - Reduced blood loss by aprotinin in thoracic surgical operations associated with high risk of bleeding. A placebo-controlled, randomized phase IV study. AB - OBJECTIVE: Although the blood-saving effect of aprotinin has been well documented in cardiac surgery and lung transplantation, its use in lung surgery has received less attention. We present our experience with the intraoperative application of aprotinin in lung resections with a predicted high risk of bleeding. METHODS: Thirty-eight patients undergoing major thoracic surgical procedures were randomized into treatment and placebo groups. The treatment group (n=18) received a bolus of 2 x 10(6) kallikrein inhibitor units (KIU) of aprotinin followed by 5 x 10(5) KIU/h during surgery. The placebo group (n=20) received an isotonic saline infusion instead. RESULTS: There was no significant difference between the groups concerning diagnosis, co-morbidity, age, sex, height, and weight. The mean intraoperative blood loss in the treatment group was significantly reduced (342 vs. 808 ml, P<0024), postoperative blood loss was also reduced (623 vs. 1282 ml, P<0.0007) and the need for blood transfusion was less (14 vs. 60, n.s.). No severe side effects of aprotinin were registered. Re-thoracotomy was necessary in two patients of the placebo group because of postoperative bleeding. CONCLUSION: Aprotinin reduces the perioperative blood loss and the need for blood transfusion in thoracic surgical procedures in patients with an increased risk of bleeding. PMID- 11423272 TI - Is video-assisted thoracic surgery justified at first spontaneous pneumothorax? AB - OBJECTIVE: The fundamental role of video-assisted thoracic surgery (VATS) in the treatment of spontaneous pneumothorax is generally acknowledged today. This study intends to evaluate whether VATS is justified at the onset of a first spontaneous pneumothorax through analysis of parameters tested on two group of patients treated respectively with pleural drainage and VATS. PATIENTS/METHODS: The study includes 70 patients affected by first spontaneous pneumothorax divided into two groups of 35 patients for the purpose of therapeutic treatment. The first group underwent pleural drainage while the second underwent VATS. Parameters analyzed were as follows: (1) prolonged air leaks (more than 6 days); (2) time required for pleural drainage; (3) time of hospital stay; (4) management costs; (5) recurrences (follow-up at 12 months). RESULTS: Prolonged air leaks occurred in four patients (11.4%) in the first group and two patients (5.7%) in the second; recurrences occurred in eight patients in the first group (22.8%), and only one in the second group (2.8%). Mean time for drainage and hospitalization was, respectively, 9 and 12 days in patients with pleural drainage against 3.9 and 6 days of those using VATS. Average management costs per patients including hospitalization was calculated at $2,750.00 per patient for the first group compared with $1,925.00 for the second group. CONCLUSIONS: The use of VATS at first spontaneous pneumothorax is justified in the interest of both patients and health administrations as demonstrated by the number of recurrences in patients in the first group and economy savings resulting from use of VATS. PMID- 11423273 TI - Acute major airway injuries: clinical features and management. AB - OBJECTIVE: Patients with an acute major airway injury are coming at our attention with increasing frequency. Despite of its nature, post-traumatic or iatrogenic, these lesions may be life-threatening. An early diagnosis and a prompt treatment reduce morbidity and mortality. MATERIALS AND METHODS: In the last 10 years, on a total of 55 patients treated in our institution for benign lesions of the major airway, 20 were with an acute injury; eleven females and nine males with a mean age of 58 years (range of 24--92). Twelve lesions were iatrogenic (orotracheal intubation) and eight were post-traumatic (three blunt traumas, five penetrating traumas). The cervical trachea was involved in 13 cases (one associated to an incomplete esophageal transection and two associated to laryngeal injuries), the thoracic trachea in six cases (four extended to the right mainstem one and to the left). Sixteen patients underwent immediate surgical repair (13 direct sutures of the tear and three complex restorations of the airway): 11 by cervicotomy and five by thoracotomy. In six cases the suture of a posterior tracheal wall tear was achieved through a new approach which provides for a small collar incision and a longitudinal tracheotomy. RESULTS: All the patients were discharged healed with a normal patency of the airway. At a mean follow up of 49 months (range of 9 -122) endoscopy showed a perfect healing process of the lesions. One patient, treated in a conservative fashion, required endoscopic laser Nd-YAG removal of a granuloma. CONCLUSION: Early diagnosis and surgical repair are the goals to persecute to achieve the best outcomes in this potentially lethal lesions. The surgical approach should be the thoracotomy if the trauma involves the 1/3 inferior trachea and/or a mainstem, the cervicotomy in the case it was injured the 2/3 superior trachea and the larynx. Posterior tracheal wall tears may be repaired via the new transcervical/transtracheal technique. The conservative treatment should be reserved to those patients with minimal signs and symptoms, and with an adequate patency of the airways. PMID- 11423274 TI - Diaphragmatic injuries. AB - OBJECTIVES: (1) To determine the actual incidence rate of blunt and penetrating diaphragmatic injuries (DI); (2) to evaluate the effectiveness of urgent surgical intervention for treatment of DI; and (3) to reveal main causes of postoperative complications. METHODS: We reviewed: (1) forensic medical examination charts of 3353 subjects, who died due to polytrauma (including injuries to the chest and/or abdomen) at accident sites; and (2) medical case reports of 4857 patients, treated for thoracoabdominal trauma (TAT) from 1962 to 1998. A detailed analysis was completed with 12 years (1987--1998) of clinical experience, involving 65 (43 penetrating, and 22 blunt) cases of DI. RESULTS: According to forensic medical data, blunt and penetrating DI occurred in 3.7% and 2.6% of individual cases, respectively. Among patients suffering from TAT, it was revealed that blunt DI had occurred in 1.1%, and penetrating in 3.9% of the cases. This data indicates if all the victims, who had sustained TAT, had survived, the incidence rate of DI would have been 2.6% (blunt -- 2.1%, and penetrating -- 3.4%). All the patients, provided surgical operations due to DI, survived. Morbidity in patients, suffering from blunt and penetrating DI, was 50%, and 35%, respectively. In the group of patients, suffering from penetrating DI, shock, intrapleural and/or intraabdominal haemorrhage, and liver injuries constituted a significant (P<0.05) influence, relevant to development of postoperative complications. The risk of complications was significantly (P<0.05) greater in cases of gunshot injuries. Fractures of chest bones, injuries of abdominal organs, and intraabdominal haemorrhage constituted a significant influence (P<0.05), relevant to development of complications after blunt DI. CONCLUSIONS: (1) The danger to the health or even life of patients is not directly caused by DI, but by consequential complications and associated injuries; (2) the effectiveness of treatment is determined by purposeful surgical diagnostics with particular regard to DI and urgent surgical intervention. PMID- 11423275 TI - Rewarming from accidental hypothermia by extracorporeal circulation. A retrospective study. AB - OBJECTIVE: Twenty-six patients with accidental hypothermia combined with circulatory arrest or severe circulatory failure were rewarmed to normothermia by use of extracorporeal circulation (ECC). The aim of the present study was to evaluate our results. PATIENTS AND METHODS: The treatment of six female and 20 male patients (median age: 26.7 years; range 1.9--76.3 years) rewarmed in the period 1987--2000 was evaluated retrospectively. Hypothermia was related to immersion/submersion in cold water (n=17), avalanche (n=1) or prolonged exposure to cold surroundings (n=8). Prior to admission, the trachea was intubated and cardiopulmonary resuscitation (CPR) initiated in all patients with cardiorespiratory arrest (n=22), whereas in those with respiration/circulation (n=4) only oxygen therapy via a face mask was given. RESULTS: Nineteen of the 26 patients were weaned off ECC whereas seven died because of refractory respiratory and/or cardiac failure. Eight of the 19 successfully weaned patients were discharged from hospital after a median of 10 days. One patient died 3 days after circulatory arrest (complete atrioventricular block) resulting in severe cerebral injury. The remaining ten patients died following 1--2 days due to severe hypoxic brain injury (n=5), cerebral bleeding (n=1) or irreversible cardiopulmonary insufficiency (n=4). Based on the reports from the site of accident, two groups of patients were identified: the asphyxia group (n=15) (submersions (n=14); avalanche accident (n=1)) and the non-asphyxia group (n=11) (patients immersed or exposed to cold environment). Seven intact survivors discharged from hospital belonged to the non-asphyxia group whereas one with a severe neurological deficit was identified within the asphyxia group. CONCLUSION: Patients with non asphyxiated deep accidental hypothermia have a reasonable prognosis and should be rewarmed before further therapeutic decisions are made. In contrast, drowned patients with secondary hypothermia have a very poor prognosis. The treatment protocol under such conditions should be the subject for further discussion. PMID- 11423276 TI - Neuroprotective effect of regional carnitine on spinal cord ischemia--reperfusion injury. AB - OBJECTIVE: The purpose of this study was to investigate the effect of regional infusion of carnitine on spinal cord ischemia--reperfusion (I--R) in rabbits. METHODS: The 36 rabbits were divided into four equal groups, group I (sham operated, no I--R injury), group II (control, only I--R), group III (I- R+intraaortic lactated Ringer's, LR, during aortic occlusion), group IV (I--R+LR plus 100mg/kg carnitine). Spinal cord ischemia was induced by clamping the aorta both below the left renal artery and above the aortic bifurcation. The spinal cord function of all animals was assessed clinically 24h after aortic declamping. Spinal cord samples were taken to measure the levels of tissue malondialdehyde (MDA) and to evaluate the histopathological changes. RESULTS: We found significant increases in the levels of MDA in groups II and III compared with group I (P<0.01), and elevation of MDA in group IV was insignificant. In group II, all animals (100%) were paraplegic with Tarlov's score of 0 and in group III, eight animals (88%) were paraplegic with Tarlov's score of 0 or 1. None of the animals (0%) from group IV was paraplegic. Histologic examination of spinal cords from group IV animals revealed that the appearance of the spinal cord was relatively preserved, whereas spinal cords from groups II and III had evidence of acute neuronal injury. CONCLUSION: The results suggest that regional infusion of carnitine during aortic clamping reduces spinal cord injury and prevents neurologic damage in rabbit spinal cord I--R model. PMID- 11423277 TI - Approach to sinus of Valsalva aneurysms: a review of 53 cases. AB - OBJECTIVE: The reported experience with sinus of Valsalva aneurysms (SVAs) is limited. Our approach to this subset of patients and an algorithm-dependent classification are presented. METHODS: Between 1985 and 2000, 53 patients (mean age: 24+/-12; range 4--60) underwent repair for ruptured (64%) or non-ruptured (36%) SVA. Associated lesions were present in 21 patients; VSD in 18, moderate to severe aortic insufficiency in five, aortic stenosis in four (two subaortic membrane and one bicuspid valve), PDA in two, mitral insufficiency in one, tetralogy of Fallot in one and endocarditis in one. Operative procedures included simple or Teflon pledgetted direct suturing (31 cases; 58%), patch repair (21 cases; 40%), and stentless porcine bioprosthetic aortic root replacement in a case with extensive involvement and aortic root distortion (2%). Concomitant procedures were VSD repair in 18 patients, aortic valve replacement in four, aortic valve resuspension in three, subaortic membrane resection in two, PDA ligation in two, mitral annuloplasty in one and total correction in one. RESULTS: Early mortality was 1.9%. A permanent pacemaker was inserted in one patient due to complete heart block. The survivors were followed up for 8.2+/-5 years (range: 21 days to 15 years). There were three reoperations due to suture dehiscence; patch repair was undertaken in these patients with no further unfavorable consequences. All patients were in NYHA Class I or II as of their last follow-up. CONCLUSIONS: Repair of SVA can be performed with an acceptably low operative risk and a good symptom-free long-term outcome expectation. Echocardiography provides all the necessary details for diagnosis. Dual exposure/patch repair strategy is advocated in the ruptured cases. PMID- 11423278 TI - Valve sparing aortic root reconstruction versus composite replacement -- perioperative course and early complications. AB - OBJECTIVE: In patients with aneurysm of the ascending aorta, dilatation of the sinotubular junction is the major cause of aortic valve regurgitation. Valve sparing aortic root replacement in patients without valvular structural defects offers a new form of treatment. The aim of this study was the assessment of the perioperative course and early complications of this method compared to composite replacement in a large single center cohort. METHODS: From 1992 to 1999, valve sparing replacement of the ascending aorta (recon) has been performed in 78 patients, while 269 patients underwent replacement by a composite graft (comp). A comparison of matched pairs (n=52) with respect to age, gender, presence of Marfan's syndrome, aortic dissection as well as date of surgery, was chosen. Aortic insufficiency was 2.8+/-0.7 for recon vs. 2.2+/-1.1 for comp preoperatively. Course and length of hospitalization, echocardiographic follow up, complications, and mortality were compared at 1-year follow-up. RESULTS: There were no operative deaths. During follow-up, one patient (2%) died 5 months postoperatively (recon) vs. two patients (3.9%) in the comp group. Bypass-time (123+/-31 vs. 153+/-31 min, P<0.0001) and cross-clamp-time (82+/-22 vs. 120+/-23 min, P<0.0001) were significantly shorter in comp. Stay in ICU (1.9+/-1.6 for recon vs. 2.3+/-2.1 days for comp) and post-op hospitalization (18.3+/-5.7 vs. 21.2+/-11.1 days) were comparable. Improvement of NYHA-class was significant after both operations (recon 2.6+/-0.8 vs. 1.3+/-0.5 and for comp 2.4+/-0.6 vs. 1.5+/-0.7, both P<0.0001). One patient (1.9%) in the recon group had to be reoperated for valve failure. Thrombembolic or bleeding complications were observed in 6 patients (12%) in comp, zero in recon (P=0.027). CONCLUSION: Valve sparing aortic root reconstruction is feasible with low perioperative morbidity and mortality and good early results. Major advantages of recon are significant reduction of thrombembolic and anticoagulation related complications as opposed to longer cross-clamp and bypass times as well as a valve failure in one patient. Further follow-up is needed to confirm our data in a long-term perspective. PMID- 11423279 TI - Fate of the aortic root after arterial switch operation. AB - OBJECTIVE: Concerns have been voiced about possible dilation and insufficiency of the neo-aortic valve after the arterial switch operation (ASO). AIMS: To determine growth of the neo-aortic valve and the aortic anastomosis after ASO and the prevalence of insufficiency or stenosis. PATIENTS AND METHODS: Since 1977, 144 consecutive patients (pts) underwent ASO for transposition of the great arteries (TGA). Median follow-up was 8.65 years (0.1--22.5 years). Simple TGA was present in 97 pts and 47 had TGA with ventricular septal defect (VSD). Detailed echocardiography included measurements of aortic diameter at four levels. The 608 measurements were compared with published normal values. RESULTS: The mean aortic valve z-score was 1.5, without significant change with age (P=0.75). Under 4 months, mean valve z-score was 0.63+/-2.20, between 5 and 12 months 2.56+/-2.30 (P<0.0001). Gradual growth occurs thereafter. The aortic sinus follows an identical growth pattern. The aorta at the anastomosis, is initially smaller than normal (z-score -0.64). After 4 months the z-score is 0.83, followed by continued growth of 0.1 z-score per year. At the last visit, the aortic valve z-score was above 2 in 51 patients, between -2 and 2 in 72 and less than -2 in six patients, none of whom had a flow velocity above 2 m/s. z-score of patients with VSD remained above those without VSD (P<0.0001). Aortic insufficiency was grade 2/4 in three patients, grade 3/4 in one and grade 4/4 in one. No patient developed aortic stenosis. CONCLUSION: After ASO the neo-aortic valve and sinus are larger than normal, representing the natural size difference in the prenatal situation and influence of associated cardiac malformations. In the first year of life, rapid dilatation of the new aorta is observed, followed by growth towards normalization of the valve and sinus size. Stenosis at the anastomosis was not observed. Aortic dilatation by itself is rarely associated with significant insufficiency. PMID- 11423280 TI - Is there a place for pediatric valvotomy in the autograft era? AB - OBJECTIVE: Valvotomy and the autograft procedure are the most common surgical treatment options for children with valvular aortic stenosis. We evaluated the results of these surgical procedures in our institution. METHODS: Retrospective analysis was done of all patients presenting with aortic stenosis and operated upon before the age of 18. In 11 patients a valvotomy was performed and in 36 an autograft procedure. RESULTS: There was no hospital mortality. Mean follow-up in the valvotomy group was 4.8 years (SD 3.3), in the autograft group 4.5 years (SD 3.3). During follow-up one patient died suddenly 2 months after valvotomy. Two patients in the autograft group died (not valve-related). After valvotomy three patients underwent a balloon valvotomy, in one followed by an autograft procedure and one patient had a repeat valvotomy. In the autograft group one patient was reoperated for severe aortic regurgitation and moderate pulmonary stenosis. At last echocardiography after valvotomy (eight remaining patients) in only two patients (25%) no aortic stenosis or regurgitation was present. In the remaining six patients aortic stenosis is mild in two and moderate in three, including one with moderate aortic regurgitation. In one patient without stenosis, moderate aortic regurgitation was seen. No pulmonary stenosis or regurgitation is present. Echocardiography after autografting (33 remaining patients) showed no aortic stenosis. Aortic regurgitation was mild in seven patients, moderate in two, severe in one. Pulmonary stenosis was present in two patients (16%). Pulmonary regurgitation was mild in three patients and moderate in one. CONCLUSIONS: In selected patients with valvular aortic stenosis who are beyond infancy, valvotomy may be adequate and may postpone further surgery for a significant length of time. After valvotomy the main problem is residual aortic stenosis while after autografting a shift occurs to aortic regurgitation and problems related to the pulmonary valve. Careful clinical and echocardiographic follow-up is therefore warranted in young patients after the autograft procedure. PMID- 11423281 TI - Repair of truncus arteriosus: a considered approach to right ventricular outflow tract reconstruction. AB - OBJECTIVE: In repair of truncus arteriosus the accepted methods of establishing right ventricle (RV) to pulmonary artery (PA) continuity utilize an allograft or xenograft valved conduit. Alternatively, the PA confluence may be directly anastomosed to the RV with anterior patch augmentation, which may allow growth and delay or avoid subsequent RVOT obstruction. These methods of RVOT reconstruction were evaluated in infants undergoing truncus arteriosus repair. METHODS: A retrospective analysis of 61 infants undergoing repair of truncus arteriosus between November 1988 and June 2000 was performed. Median age was 34 days (range 1 day to 6.4 months). The patient cohort was subdivided into two groups (1) Valved conduit group: RV to PA continuity performed with a conduit in 38 patients using allograft (28) or xenograft (10). (2) Direct anastomosis group: direct RV-PA anastomosis performed in 23 patients, augmented anteriorly with monocusp (15) or simple pericardial patch (eight). RESULTS: There were eight hospital deaths (13%, 95% confidence limits 5--21%). Hospital mortality did not differ significantly between group 1 and 2 (three patients (8%) versus five patients (22%) respectively, P=0.23). By multivariate analysis, low operative weight (P=0.023), severe truncal regurgitation (P=0.022) and major coronary abnormalities (P=0.018), were independent risk factors for hospital death. Hospital survivors were followed-up from 1.3 months to 11.8 years (mean 4.2+/-3.4 years). There were eight late deaths with survival of 73+/-6% at 2 years and beyond. Survival was not influenced by method of RVOT reconstruction (Conduit versus direct RV-PA anastomosis, 2.76+/-7%, 63+/-10%, respectively, P=0.23). Freedom from surgical RVOT reintervention was 56+/-10% in group 1 and 89+/-10% in group 2 at 10 years (P=0.023). The use of a xenograft conduit was an independent risk factor for reintervention (P<0.001). CONCLUSIONS: In truncus arteriosus repair, RV to PA continuity established by a direct anastomosis was associated with a low incidence of surgical RVOT re-intervention. This technique has the potential for RVOT growth and may be a useful alternative when an appropriate allograft is unavailable, particularly in the neonate where the risk of pulmonary hypertension are lower. PMID- 11423282 TI - Mitral valve replacement with mechanical prostheses in children: improved operative risk and survival. AB - OBJECTIVE: The purpose of this study was to assess the early and late outcome following mitral valve replacement (MVR) with mechanical prostheses in children. PATIENTS AND METHODS: Between 1981 and 2000, 44 consecutive children (mean age 6.8+/-4.7 years, 2 months--16 years) underwent mechanical MVR in Southampton. Twenty-three children were less than 5-years-old and nine were infants. Disease aetiology was congenital in 37, rheumatic in four, infective in two and Marfan's syndrome in one. Mitral regurgitation was present in 36 and mitral stenosis in eight. Concomitant procedures were performed in 13, including aortic valve replacement (AVR) in seven. Follow-up was complete (mean 6.4+/-4.8 years, 1 month -18.1 years). RESULTS: The overall operative mortality was 14% (six patients). Before and after 1990 operative mortality was 31 vs 3.6% (P=0.02). From 1990, operative mortality for infants was zero out of six, for children less than 5 years-old was one out of 16 (one death after emergency AVR and MVR) and for older children it was 0/12. Seven children experienced valve or anticoagulation treatment-related events and eight had a mitral valve re-operation. Ten-year freedom from thromboembolism, prosthetic valve infection, bleeding, paravalvular leak and a mitral valve re-operation was 92.8+/-5.2, 97.3+/-2.7, 97.7+/-2.3, 97.2+/-2.7 and 75+/-9.7%, respectively. Overall 10-year survival was 78+/-7% (four late deaths); for children under vs over 5 years it was 61+/-11 vs 95.2+/ 4.6% (P=0.02), for atrio-ventricular septal defect (AVSD) vs other pathology 55+/ 15 vs 89+/-6.1% (P=0.05) and for those operated before 1990 vs after 1990 it was 63+/-8.1 vs 86+/-8.2% (P=0.04). CONCLUSIONS: Mechanical MVR, in the current era, carries a low operative risk across the spectrum of paediatric age. Late survival is better for older children and those having no-AVSD pathology but it has improved substantially during the 1990s irrespective of age and disease aetiology. PMID- 11423283 TI - The enhancement of hemodynamic performance in Fontan circulation using pain free spontaneous ventilation. AB - OBJECTIVE: Positive pressure ventilation is known to have a deleterious effect on pulmonary blood flow in patients with Fontan physiology. We evaluated the hemodynamic effects of pain free, spontaneous, non-positive pressure ventilation in patients undergoing Fontan staging procedures or completion. Fontan procedures, with creation of low pressure passive pulmonary circulation. METHODS: Between May 1997 and May 1999 50 consecutive patients undergoing either bi directional Glenn (BDG, n=23) or completion Fontan (n=27), were managed with early extubation. Anaesthetic management included continuous narcotics, caudal block, epidural block, or hyperbaric spinal. Post-operative management included low dose dopamine (3 mcg/kg per min), nitro-glycerine (0.3 mcg/kg per min) and nitroprusside (0.3 mcg/kg per min). Post-operative management was identical for all patients. Twelve patients were randomly selected to undergo continuous cardiac output and cardiac index (CI) determinations utilizing extra vascular Doppler probes placed on the ascending aorta, allowing for continuous aortic diameter and Doppler wave form velocity recordings. All patients were extubated either in the operating room or within one hour post-operatively. There were no deaths and no complications in the series. Mean length of stay (LOS) for BDG was 4.3+/-0.5 days. Mean LOS for Fontan patients was 11+/-4 days. RESULTS: Mean pulmonary artery pressure (MPAP) fell from 19+/-3.464 pre-extubation to 14+/ 3.271 immediately post-extubation, 13.2+/-2.261 6 h post-extubation, and 11.7+/ 2.146 12 h post-extubation. All decreases in MPAP post-extubation were significant (P=<0.05). CI pre-extubation was 3.25+/-1.09, immediately post extubation 5.05+/-1.297, 12 h post-extubation 6.225+/-1.19. All increases in CI post-extubation were significant (P=<0.05). CONCLUSION: Resumption of pain free, spontaneous, non-positive pressure ventilation enhances hemodynamic performance in patients with Fontan circulation and clearly improves outcome. PMID- 11423284 TI - Does the completeness of revascularization affect early survival after coronary artery bypass grafting in elderly patients? AB - OBJECTIVE: Usefulness and risks of incomplete versus complete revascularization are still matters of ongoing discussions. Because an increasing number of elderly patients are undergoing coronary artery bypass grafting (CABG), the question arises whether a less extensive surgical approach is more prudent than complete revascularization. METHODS: Of 6531 patients undergoing isolated CABG, 859 were 75 and older at the time of operation. Mean age of the 859 patients was 77+/-2.7 years (median: 76 years); 65% were men. Follow-up enquiry by questionnaire was performed at the 180th postoperative day with a completeness of 95.6%. Assessment of the impact of incomplete revascularization utilized both multivariable analysis and propensity score matching to account for selection factors. RESULTS: Incomplete revascularization was performed in 133 patients (16%). The most common reasons for incomplete revascularization were small vessels (55%) and massive calcification (32%). Mortality until 180 days after CABG was higher (n=32; 24%) after incomplete than after complete revascularization (n=105; 15%; P=0.005). By logistic multivariable regression, incomplete revascularization was identified as an independent risk factor for death (Odds ratio, 1.8; P=0.015). By time-related analysis, incomplete revascularization predominantly affected the early period after CABG (P=0.001). Aortic cross clamping time was only slightly shorter for the group with incomplete (59+/-27 min (median: 55 min) vs. 63+/-26 min (median: 58 min); P=0.1). CONCLUSIONS: Incomplete revascularization increases the early risk of death after CABG in patients aged 75 years and older. The potential compensating benefit of the shorter aortic cross clamping time does not outweigh the advantages of complete revascularization. Thus, in the era of high-volume interventional approaches and minimally invasive techniques, the advantages of complete revascularization need to be considered. PMID- 11423285 TI - Single-clamp technique does not protect against cerebrovascular accident in coronary artery bypass grafting. AB - OBJECTIVES: By potentially avoiding the embolic consequences of a side-biting aortic clamp, the single-clamp technique may decrease cerebrovascular accidents in coronary artery bypass grafting. However, this theoretical superiority in stroke prevention has not been conclusively demonstrated and use of this technique may lead to adverse myocardial effects due to longer cross-clamp times. In this study, we sought to determine if the single-clamp technique prevents postoperative stroke in clinical practice. METHODS: Of 607 consecutive isolated coronary bypass operations completed over a 3 year period, 301 (50%) were performed by one surgeon using exclusively the single-clamp technique and 306 (50%) were performed by a second surgeon using exclusively the two-clamp technique. Postoperative adverse events were retrospectively compared between these two groups. RESULTS: There were no differences between groups in terms of postoperative stroke (1.7% single-clamp vs. 2.0% two-clamp, P=0.78), hospital mortality (2.7% single-clamp vs. 1.6% two-clamp, P=0.38), or perioperative myocardial infarction (2.6% single-clamp vs. 0.7% two-clamp, P=0.052). The two clamp technique was not a significant predictor of stroke by logistic regression analysis (P=0.72). CONCLUSIONS: We conclude that there are no statistically significant differences between clamp techniques with regard to stroke prevention or myocardial protection. We find no compelling evidence for surgeons successfully utilizing one technique to change to the other. PMID- 11423286 TI - Fewer reoperations and shorter stay in the cardiac surgical ward when stabilising the sternum with the Ley prosthesis in post-operative mediastinitis. AB - OBJECTIVE: Using the Ley prosthesis, a 0.5mm thick titanium alloy plate for stabilising the sternum, is a new method in the treatment of mediastinitis after open-heart surgery. We report a retrospective analysis of our experience with this device. METHODS: One hundred consecutive cases of post-operative mediastinitis in the period 1992-1997 were reviewed. The primary procedure at reoperation for infection was as follows: 52 patients were treated with the Ley prosthesis and 48 patients underwent other conventional procedures. The choice of the surgical technique depended on the attending surgeon. The prosthesis was used more frequently in patients with sternal dehiscence (P<0.001) but otherwise patients' characteristics were similar in the two groups. As a control population for outcome data, 100 uninfected patients were matched with regard to operative procedure, age, sex and date of surgery. RESULTS: The median hospital stay was 48.5 days in the mediastinitis group vs. 14 days in the control group. The all cause 90-day mortality in the mediastinitis group was 18% vs. 5% in the control group. The 52 patients treated with the Ley prosthesis had a median length of stay in the cardiac surgery ward for 29 days vs. 41.5 days in the mediastinitis group not treated with the prosthesis (P=0.013). However, when the total length of stay including hospitals outside the cardiac surgery ward was taken into account, the prosthesis did not reduce the length of stay. Only 8/52 patients treated with the prosthesis required further surgery vs. 23/48 patients who were not primarily treated with the prosthesis (P<0.001). The Ley prosthesis had no impact on mortality. CONCLUSION: The Ley prosthesis is a valuable adjunct to the treatment of mediastinitis after open-heart surgery. A shorter stay at the cardiac surgery ward and a reduced need for further surgical procedures were observed when using this prosthesis. PMID- 11423287 TI - Plasma levels of atrial and brain natriuretic peptides as indicators of recovery of left ventricular systolic function after coronary artery bypass. AB - OBJECTIVE: The purpose of this study was to investigate the effectiveness of atrial and brain natriuretic peptides (ANP and BNP, respectively) as indicators of recovery of left ventricular (LV) function after coronary surgery. METHODS: We measured the concentrations of these peptides in 31 patients with poor LV function (ejection fraction, EF<35%) undergoing coronary artery bypass, and evaluated their correlation with the echocardiographic indexes of LV function. RESULTS: Pre-operatively, the plasma levels of both ANP and BNP were markedly higher in coronary patients than in normal control subjects, and strongly correlated with both EF (BNP: r=-0.8, P<0.001; ANP: r=-0.6, P<0.001) and wall motion score index (WMSI). At post-operative follow up, plasma levels of both natriuretic peptides were markedly reduced compared with pre-operative values in 21 patients. In addition, the post-operative-pre-operative differences of BNP (Delta(BNP)) and ANP (Delta(ANP)) plasma levels strongly correlated with the differences of both EF (r=-0.7, P<0.0001 vs. Delta(BNP); r=-0.6, P=0.0003 vs. Delta(ANP)) and WMSI (r=0.6, P=0.002 vs. Delta(BNP); r=0.6, P=0.04 vs. Delta(ANP)). Finally, by logistic regression analysis, BNP appeared a significant predictor of LVEF recovery after surgery. CONCLUSION: Plasma levels of ANP and BNP might be used in routine clinical practice as a support to echocardiography in detecting recovery of the LV function after coronary surgery. PMID- 11423288 TI - Protamine cardiotoxicity and nitric oxide. AB - OBJECTIVES: The purpose of this study is to assess the role of the nitric oxide (NO) pathway in protamine-induced cardiotoxicity and to formulate a possible explanation for this adverse effect. METHODS: Isolated rat hearts were perfused by Krebs--Henseleit (KH) solution using a modified Langendorff model. They were randomized into three groups: A, 40 min perfusion with KH solution; B, 20 min perfusion with KH solution and 20 min with protamine; C, as B but Ng-monomethyl-L arginine (L-NMMA), a non-selective inhibitor of the NO pathway, was added during 40 min of the perfusion period. Left ventricular (LV) function was measured every 10 min. NO and tumor necrosis factor-alpha (TNF) were detected in the effluent from the coronary sinus (CS) and in the supernatant of the cardiac myocytes culture. Nitric oxide synthases (NOS) mRNA levels were determined in groups A and B from LV samples at baseline and after 40 min of perfusion. RESULTS: We found that protamine at a dose of 12 microg/ml causes significant depression of LV function (decreased peak systolic pressure to 22.5+/-3.2% and dP/dt max to 22.9+/ 3.1%). L-NMMA did not prevent protamine cardiotoxicity. NOS mRNA was not detected from LV samples in any group. The NO in the effluent from the CS and from the supernatant of the cardiomyocytes culture was below detectable levels. However, a significant amount of TNF was measured in the effluent from the CS (108+/-17 pg/min for group B and 117+/-13 pg/min for group C) and in the supernatant of the cardiomyocytes culture (65+/-21 pg/ml). CONCLUSIONS: This study suggests that direct protamine-induced cardiotoxicity does not depend on the NO pathway. Our finding that protamine induced TNF release by cardiomyocytes can shed new light on the understanding of protamine cardiotoxicity. PMID- 11423289 TI - Efficacy of adenoviral gene transfer with manganese superoxide dismutase and endothelial nitric oxide synthase in reducing ischemia and reperfusion injury. AB - OBJECTIVE: Both superoxide dismutase (SOD), a free radical scavenger, and nitric oxide (NO), a vasodilator with anti-inflammatory properties, have been shown to protect the myocardium from reperfusion injury. They are known to interact in vivo, the influence of which on myocardial protection has not been studied. METHODS: Four groups of rats (n=7, per group) were subjected to experimental infarction following injections into the anterior wall of the left ventricle with adenoviral vector encoding beta-galactosidase (group A), eNOS (group B), Mn-SOD (group C) and both eNOS and MnSOD (group D). Hearts were assessed for protein expression and size of infarction. RESULTS: Efficiency of gene up regulation was confirmed by immunostaining for eNOS and Mn-SOD, and X-gal staining for beta-gal respectively. In B and D, overexpression of eNOS was demonstrated in cardiac myocytes in addition to that in the endothelium, while in C and D, Mn-SOD was overexpressed in mainly cardiomyocytes. Infarct size was 49.7+/-4.8% in A, and was significantly reduced in the other groups (29.8+/-2.7%, 21.8+/-2.5% and 24.9+/-2.4% in B, C and D respectively). CONCLUSION: Adenoviral gene transfer of Mn-SOD was superior to eNOS in reducing the extent of in vivo ischemia reperfusion injury in the rat heart in our model. The effect of combined application of Mn-SOD and eNOS was not different from their individual effect. PMID- 11423290 TI - Efficiency of non-viral gene delivery systems to rat lungs. AB - OBJECTIVE: Transient expression of therapeutic genes within lung allografts may modulate the pathological processes following allotransplantation. Whilst efficient gene transfer to lungs has been reported with viral vectors, their usefulness is limited on the grounds of safety. Since non-viral systems overcome many of these safety issues, our studies were designed to evaluate the efficiency of several non-viral gene delivery vectors for in vivo transfer of plasmid DNA to rat lungs via the airways. METHODS: Fischer rats (230--260 g) underwent a thoracotomy, right main bronchus occlusion and instillation of 300 microg naked or complexed DNA (pCIluci, luciferase gene/CMV promoter) to the left lung followed by ventilation for 10 min. Rats were divided into five treatment groups (n=5): (1) Glucose, (2) Naked DNA, (3) Linear polyethylenimine (PEI), (4) Branched PEI, (5) Lipid GL-67/DOPE and (6) DOTAP/cholesterol. Animals were sacrificed 24 h after gene delivery for measurement of reporter gene activity and gas exchange of the left lung. RESULTS: Linear PEI was the most efficient gene delivery vector and was significantly better than DOTAP/cholesterol (P=0.00002) and naked DNA (P=0.004). All gene delivery vectors impaired function of the transfected left lung compared with DNA alone. Of all the gene delivery vectors tested, lipid GL-67/DOPE exerted the least effect on lung function whilst DOTAP/cholesterol mediated the most adverse effect. CONCLUSION: Linear PEI was the most efficient vector for gene delivery to rat lungs in our experimental setting although it mediated a moderate impairment in lung function. Further studies are needed to evaluate whether this effect is transient. PMID- 11423291 TI - Tissue engineering of small caliber vascular grafts. AB - OBJECTIVE: Previous tissue engineering approaches to create small caliber vascular grafts have been limited by the structural and mechanical immaturity of the constructs. This study uses a novel in vitro pulse duplicator system providing a 'biomimetic' environment during tissue formation to yield more mature, implantable vascular grafts. METHODS: Vascular grafts (I.D. 0.5 cm) were fabricated from novel bioabsorbable polymers (polyglycolic-acid/poly-4 hydroxybutyrate) and sequentially seeded with ovine vascular myofibroblasts and endothelial cells. After 4 days static culture, the grafts (n=24) were grown in vitro in a pulse duplicator system (bioreactor) for 4, 7, 14, 21, and 28 days. Controls (n=24) were grown in static culture conditions. Analysis of the neo tissue included histology, scanning electron microscopy (SEM), and biochemical assays (DNA for cell content, 5-hydroxyproline for collagen). Mechanical testing was performed measuring the burst pressure and the suture retention strength. RESULTS: Histology showed viable, dense tissue in all samples. SEM demonstrated confluent smooth inner surfaces of the grafts exposed to pulsatile flow after 14 days. Biochemical analysis revealed a continuous increase of cell mass and collagen to 21 days compared to significantly lower values in the static controls. The mechanical properties of the pulsed vascular grafts comprised supra physiological burst strength and suture retention strength appropriate for surgical implantation. CONCLUSIONS: This study demonstrates the feasibility of tissue engineering of viable, surgically implantable small caliber vascular grafts and the important effect of a 'biomimetic' in vitro environment on tissue maturation and extracellular matrix formation. PMID- 11423292 TI - Downregulation of myocardial contractility via intact ventriculo--arterial coupling in the brain dead organ donor. AB - OBJECTIVE: To test the hypothesis that altered loading conditions play a key role in hemodynamic instability and cardiac dysfunction in the brain dead (BD) organ donor. METHODS: BD was induced by inflation of a subdural balloon catheter. In the first part of the study, left ventricular function was assessed in a canine in situ cross-circulated heart model (n=6). Pre- and afterload and coronary perfusion pressure were kept identical in all hearts throughout the experiment. In the second part of the study, hearts (n=6) were investigated in vivo allowing the interaction between left ventricular contractility and arterial load. Left ventricular pressure--volume loops were obtained by a combined conductance pressure catheter and the slope of the endsystolic pressure--volume relationship (Ees), arterial elastance (Ea), stroke work (SW), pressure--volume area, ventriculo--arterial coupling ratio (VAC) and mechanical efficiency (Eff) were calculated. RESULTS: Induction of BD led to a hyperdynamic response in both models with a significant increase of most hemodynamic parameters. In the in situ isolated heart model, left ventricular contractility returned to baseline without any further deterioration. In contrast, in the intact circulation the hemodynamic parameters declined significantly in comparison to baseline 4 h after BD (Ees: 4.07+/-0.51 vs. 8.06+/-1.09 mmHg/ml, P<0.05, Ea: 3.17+/-0.39 vs. 4.42+/-0.30 mmHg/ml, P<0.05). However, VAC (0.78+/-0.09 vs. 0.65+/-0.14 n.s.) and Eff (73.4+/ 2.1 % vs. 76.8+/-3.7 %, n.s.) remained constant over the time. CONCLUSION: BD induction leads to an initial hyperdynamic reaction followed by hemodynamic instability. The facts that no cardiac dysfunction occurred if loading conditions were kept constant and the ventriculo--arterial coupling ratio and mechanical efficiency remained constant in the intact animal model indicate that decreased contractility reflects to decreased arterial elastance after brain death. Therefore, reduced contractile function after brain death at a decreased afterload may contribute to stroke work optimization. PMID- 11423293 TI - Transforming growth factor beta and myocardial dysfunction following heart transplantation. AB - OBJECTIVE: We analyzed the role of transforming growth factor-beta (TGF-beta), a fibrogenic cytokine, in the development of left ventricular diastolic dysfunction following heart transplantation. METHODS: We studied 152 heart transplant recipients who had survived for at least 24 months. We compared histopathological findings (staining of endomyocardial biopsy specimens using Hematoxylin Eosin and polyclonal antibodies), left ventricular function (Doppler echocardiography) and clinical course (NYHA status). Patients are classified into group A (n=56 recipients) with immunohistochemical TGF-beta staining score >7 and group B (n=96 recipients) with a staining score <7. RESULTS: Doppler echocardiographic evaluation demonstrated greater impairment of left ventricular diastolic function in recipients with higher TGF-beta staining score. The average mitral deceleration time was 129+/-6 ms for recipients group A compared to 167+/-15 ms in group B. While the mean isovolumic relaxation time was 65+/-8 ms for patients in group A compared with 82+/-6 ms for recipients in group B (P=0.0004 and 0.005, respectively). Immunohistochemical scoring correlated inversely with both mitral deceleration and isovolumic relaxation times (r=-0.74, P=0.0004 and r=-0.66, P=0.004, respectively). Mean NYHA status was 2.7+/-1.3 for group A compared to 1.17+/-0.4 in group B was (P=0.002). Five years follow-up revealed persistent left ventricular diastolic impairment for recipients with higher immunohistochemical staining score. Mitral deceleration time and isovolumic relaxation time were 118+/-11 and 62+/-7 ms for group A compared to 156+/-12 and 80+/-5 ms for group B, P=0.006 and P=0.01, respectively. The actuarial development of subsequent coronary artery disease (> 50% stenosis) was 17 and 29% for recipients in group A compared to 4 and 6% for recipients in group B at 3 and 5 years follow-up, respectively (P=0.01 and P=0.005, respectively). CONCLUSIONS: TGF-beta expression in cardiac allografts is associated with impaired graft function and limited survival. The pathogenesis of diastolic dysfunction may be an aberrant repair process following rejection due to increased TGF-beta expression in transplant recipients. PMID- 11423294 TI - Functional assessment of non-heart-beating donor lungs: prediction of post transplant function. AB - OBJECTIVES: To enable an increase in the numbers of donor lungs using organs from non-heart-beating donors (NHBD). To develop an isolated ventilation and perfusion technique to assess the degree of warm ischaemic organ injury suffered prior to retrieval, thereby enabling identification of lungs with predictably good post transplant function. METHODS: Lungs from Landrace-Yorkshire White cross pigs were retrieved after 1 (NHBD(1)), 2 (NHBD(2)) or 4 h (NHBD(4)) post-hypoxic death induced by cessation of ventilation. Control organs were retrieved using standard techniques for each group from matched animals immediately following aortic cross clamping (Control(1) and combined Control(2,4)). Modified Euro-Collins pulmoplegia was used in all groups, prior to ventilating a single lung with 100% oxygen and perfusion with neutrophil-depleted and deoxygenated blood. For all of the lungs in the NHBD(2) and combined Control(2,4) groups, and one of the successfully perfused NHBD(4), the contralateral lung was then transplanted with post-transplant function assessed for 12 h. All animals were anaesthetized throughout and euthanased without regaining consciousness. RESULTS: On assessment, oxygenation after 5 min of perfusion did not differ between NHBD(1) (n=4) vs. Control(1) (n=5; analysis of variance (ANOVA), P=0.152). However, oxygenation had deteriorated significantly in the NHBD(2) group (n=6) vs. Control(2,4) (n=8; ANOVA, P<0.0005) and was significantly poorer than initial values after 8 min (unpaired t-test with Bonferroni correction, P<0.03). In NHBD(4) (n=6), four lungs failed assessment due to the development of gross pulmonary oedema, although the remaining pair functioned as well as Control(2,4). Post-transplantation, NHBD(2) (n=6) contralateral lungs showed significantly poorer overall oxygenation, (mean+/-SD, 46+/-22 kPa) when compared with Control(2,4) (n=6; 59+/-16 kPa; ANOVA, P=0.001), although oxygenation was satisfactory. The contralateral organ from one successfully perfused NHBD(4) lung functioned well post-transplantation. CONCLUSIONS: The significant deterioration in oxygenating performance seen during assessment after 2 h warm ischaemia and the idiosyncratic function after 4 h warm ischaemia indicates the importance of functional testing of NHBD lungs. The similar deterioration in oxygenating performance seen post-transplantation in the contralateral lungs suggests that this method detects functional warm ischaemic lung injury. PMID- 11423295 TI - Freely mobile right-sided atrial thrombus mimicking myxoma. PMID- 11423296 TI - Pulmonary arteriovenous fistula. PMID- 11423297 TI - Acute pulmonary embolism due to multiple hydatid cysts. AB - A case of acute pulmonary embolism due to multiple hydatid cysts is reported. Chest X-ray, echocardiography, spiral CT scan and MR-angiography were performed for the diagnostic evaluation. The patient underwent a left anterior thoracotomy and a left pulmonary arteriotomy in order to remove the hydatid cysts, without using extracorporeal circulation. The post-operative recovery was uneventful and the patient, 42 months later, has a normal life. PMID- 11423298 TI - Papillary muscle rupture and pericardial injuries after blunt chest trauma. AB - Non-penetrating cardiac trauma resulting in mitral valve rupture is uncommon, requiring a high degree of suspicion for diagnosis. Sudden and severe mitral regurgitation, unless surgically corrected rapidly lead to congestive heart failure and death. We report a patient with traumatic rupture of the antero lateral papillary muscle of the mitral valve and pericardial injury, after a lateral blunt chest trauma, who successfully underwent emergency mitral valve replacement. PMID- 11423299 TI - Sternal dehiscence after cardiac surgery and ACE inhibitors [correction of ACE type 1 inhibition]. AB - We report two cases, which underwent surgery through Median sternotomy. They were on ACE inhibitors [corrected] pre-operatively. Both of these patients developed persistent dry cough post-operatively, which resulted in sternal wound dehiscence. They had no clinical or bacteriological evidence of sternal wound infection. Although one patient was overweight and had moderately impaired left ventricular function, there were no other associated risk factors. Both patients underwent rewiring of the sternum. Type II receptors inhibitor were introduced post-rewiring, which cured the persistent dry cough. Both the patients are enjoying a good quality of life at 2 year 6 months and 2 years post-rewiring of the sternum. PMID- 11423300 TI - Video-assisted thoracoscopic treatment for pleuroperitoneal communication in peritoneal dialysis. AB - Massive hydrothorax is an uncommon but well-recognized complication of continuous ambulatory peritoneal dialysis (CAPD). We performed a video-assisted thoracoscopic resection of the pleuro-peritoneal communication and pleurodesis in a patient with massive right hydrothorax secondary to CAPD. Histologically, the resected diaphragm was lacking in common tissue, tendons and skeletal muscle tissues, is displaced to fibrous connective tissue. These anatomic findings suggested that the cause of communication was congenital diaphragmatic change. Video-assisted thoracoscopic treatment facilitated efficient inspection and easy resection of the weak portion of the diaphragma in the case of pleuroperitoneal communication. PMID- 11423301 TI - Juxtacardiac costal osteochondroma presenting as recurrent haemothorax. AB - A 21-year-old man presenting with a recurrent spontaneous haemothorax was found to have an osteochondroma arising out of the left 4th rib. This was penetrating the apical part of the heart. Surgical excision was uneventful. PMID- 11423302 TI - Lung cancer occurring with Mycobacterium xenopi and Aspergillus. AB - Two cases which associate Mycobacterium xenopi pulmonary infection, aspergilloma and preoperative unsuspected lung cancer are related. To our knowledge, the association of these three pathologies has never been previously reported. These two cases, suggest that infected chronic lung lesions, especially in smokers, should be closely monitored and be surgically removed in order to prevent further complications. PMID- 11423303 TI - Lipid reserves used by pueruli of the spiny lobster Jasus edwardsii in crossing the continental shelf of New Zealand. AB - Lipid is known to fuel the movement of the nektonic puerulus stage of the spiny lobster Jasus edwardsii across the continental shelf of New Zealand. Lipid class analysis of pueruli caught from two locations across the continental shelf showed that phospholipid predominated (86-96% of total lipid), with only smaller proportions of sterol (0.9-8.7%) and diacylglycerol (1.2-7.6%). Only traces of triacylglycerol, hydrocarbon and wax ester were present (<0.1% of total lipid). Comparison of the lipid class content of pueruli caught onshore and offshore showed that phospholipid reserves are primarily utilised during this important phase in the lifecycle and that diacylglycerol plays a less significant secondary role. Histology identified concentrations of phospholipid in fat bodies located in the haemocoel. The use of phospholipid as the dominant storage medium in the puerulus stage is unlike many other marine taxa, including crustacea, which tend to use triacylglycerol and wax ester. The use of phospholipid as a storage medium may well be related to its characteristic transparency, an important feature of this nektonic stage of lobster development that is highly vulnerable to pelagic visual predators. PMID- 11423304 TI - Utilisation of lipids, protein, ions and energy during embryonic development of Australian oviparous skinks in the genus Lampropholis. AB - The contents of eggs and neonates of the Australian skinks, Lampropholis guichenoti and L. delicata, are described and compared to allow interpretation of nutrient utilisation by the developing embryo. Even though the females are the same size, L. guichenoti lay smaller clutches of larger eggs (egg contents=41.6+/ 1.2 mg dry mass) than L. delicata (26.6+/-2.8 mg). The energy density is the same for eggs (30.5+/-0.9 J/g ash-free dry mass for L. guichenoti and 29.9+/-1.1 J/mg for L. delicata) and neonates (22.5+/-1.3 J/mg for L. guichenoti and 23.5+/-0.4 J/mg for L. delicata) between species. The amount of nitrogen (protein) in neonates is only slightly lower than that in eggs, whereas there is a large and significant decline in total lipids. Thus, like some other skinks, protein is a source of metabolic energy during embryogenesis, although not as important as lipid. Triacylglycerol is the major lipid component of the eggs (80% of total lipid), with phospholipid forming only approximately 10% of the total lipid. The fatty acid profile of the phospholipid is distinguished by a high proportion of arachidonic acid (8%), a significant proportion of eicosapentaenoic acid (2-4%) and a relatively low proportion of docosahexaenoic acid (2-3%) compared to chickens. Eggs of both species have remarkably low concentrations of free cholesterol compared to other amniote eggs (0.7% for L. guichenoti and 1.3% for L. delicata). The loss of lipid during embryonic development is almost entirely due to the selective utilisation of yolk triacylglycerol, presumably for energy. By contrast, the amount of phospholipid recovered from the neonates was the same as that originally in the eggs. Moreover, significantly more total cholesterol was present in the neonates than in the eggs, suggesting that biosynthesis of additional cholesterol occurred during development. The phospholipids of the neonates contain higher proportions of arachidonic (11-12%) and docosahexaenoic (8%) acids than the phospholipids of the eggs. Eicosapentaenoic acid is less prevalent in phospholipids in neonates than in eggs. Neonates of both species contain significantly more calcium than the fresh egg contents (L. guichenoti, eggs 0.303+/-0.051 mg, neonates 0.641+/-0.047 mg; L. delicata, eggs 0.187+/-0.013 mg, neonates 0.435+/-0.033 mg), presumably as a result of resorption of calcium from the eggshell. Interestingly, there is also significantly more sodium in neonates than in the contents of fresh eggs (L. guichenoti, eggs 0.094+/-0.010 mg, neonates 0.184+/-0.011 mg; L. delicata, eggs 0.084+/-0.011 mg, neonates 0.151+/-0.010 mg). There is no significant difference in the content of potassium and magnesium in eggs and neonates of either species. Although the fresh eggs of L. delicata have a significantly higher sodium concentration than L. guichenoti, there is no difference in the concentrations of calcium, magnesium, potassium or sodium in the neonates of the two species. PMID- 11423305 TI - Expression of messenger RNA for gonadotropin receptor in the granulosa layer during the ovulatory cycle of hens. AB - The present experiments were conducted to evaluate the mRNA levels of luteinizing hormone receptor (LHR) and follicle-stimulating hormone receptor (FSHR) in granulosa layers during the ovulatory cycle of hens, in relation to the release of LH and steroid hormones. After the release of LH, progesterone (P4) and estradiol-17beta (E2), found 4-5 h before ovulation, LHR and FSHR mRNA levels were observed to decrease in the granulosa layers of the largest (F1) and second largest (F2) preovulatory follicles, with the greatest in the LHR mRNA level of F1. P4 concentrations in the granulosa layers of F1 and F2 increased 4-5 h before ovulation, with greater in F1 than in F2. F2 concentrations in the theca layers were greater in F2 than in F1 throughout the ovulatory cycle. Also, the injection of ovine LH caused decreases in the mRNA levels of LHR and FSHR in the granulosa layers. However, these decreases were abolished by the injection of aminoglutethimide, an inhibitor of steroid synthesis. These results suggest that in hen granulosa cells, the mRNA levels of not only LHR but also FSHR are down regulated by LH and the down-regulation may be mediated steroid hormones. PMID- 11423306 TI - An allometric analysis of oxygen consumption rate and cardiovascular function in the cockroach, Blaberus discoidalis. AB - The interrelationship of metabolic rate and cardiovascular function has been well documented in vertebrates through allometric analyses. However, similar studies are lacking in insects. Unlike vertebrates, the cardiovascular system of insects does not play a significant role in oxygen transport. A comparison of the interrelationship in insects and vertebrates might provide insight into the nature of the connection between metabolic rate and the cardiovascular system. Oxygen consumption, heart rate and heart dimensions were measured in the nymphs of the cockroach Blaberus discoidalis over a mass range of 0.03-5 g. Oxygen consumption rate scaled with an exponent of 0.83. The relationship between heart rate and body mass scaled negatively, however, it did not appear to be linear. Using measurements of heart widths, abdominal length and heart rate, stroke volume and cardiac output were estimated. Cardiac output appeared to scale linearly with an exponent of 0.85, which was not significantly different from the exponent observed for the rate of oxygen consumption. Thus, the observed similarity between the exponents for oxygen consumption rate and cardiac output in vertebrates also appears to be present in insects. PMID- 11423307 TI - Water-soluble luminal contents of the gut of the earthworm Lumbricus terrestris L. and their physiological significance. AB - In the post-gizzard gut of the earthworm Lumbricus terrestris, distinguishing the functions of the luminal epithelium from those of the chloragogenous tissue has been hindered by the close apposition of these two tissues. Moreover, both tissues may have different functions from the anterior to the posterior of the animal. We analyzed the gut luminal contents of L. terrestris so as to gain a better understanding of the function of the luminal epithelium. The intestine was divided into four regions from anterior to posterior, and the water-soluble portion of the luminal contents of these four regions was analyzed for protease and amylase activity, calcium and ammonium ions, and protein. The same four regions of the gut wall were analyzed for glutamate dehydrogenase (GDH) and serine dehydratase (SDH) to determine their location with reference to the site of ammonia production. We observed high levels of proteases, amylase, protein and calcium ions in the gut luminal contents of the first two regions, and a significant decline of all four variables in region III. Conversely, ammonia was low in the gut contents of regions I and II but rose sharply in region III, which was also the region to which the tissue enzymes GDH and SDH were localized. The ammonia content of earthworm casts was observed to be much higher than that of the surrounding soil. These data are presented as partial evidence for the proposal that the excretory ammonia produced by feeding earthworms is a product of the luminal epithelium of region III of the gut. It is also proposed that ammonia and calcium may function as ion-exchangers in the absorptive function of the earthworm gut. PMID- 11423308 TI - Effect of phospholipase C, trypsin and neuraminidase on binding of bilirubin to mammalian erythrocyte membranes. AB - Binding of bilirubin to erythrocyte membranes of human, buffalo, sheep and goat was studied after phospholipase C, trypsin and neuraminidase treatment. Phospholipase C and trypsin treatment of membranes greatly enhanced the bilirubin binding in all mammalian species, whereas, neuraminidase treatment resulted into a small increase in the membrane-bound bilirubin. Human erythrocyte membranes bound the highest amount of bilirubin, whereas buffalo, sheep and goat erythrocyte membranes showed different mode of bilirubin binding. The order of bilirubin binding to unmodified as well as neuraminidase-treated erythrocyte membranes was: human>sheep>buffalo>goat; the order was: human>buffalo>sheep>goat; in phospholipase C- and trypsin-treated erythrocyte membranes. These binding results indicate that membrane phospholipids are directly involved in the interaction of bilirubin with the membranes as the differences observed in the membrane-bound bilirubin among mammalian species were directly correlated with the sum of choline phospholipids, especially phosphatidylcholine and sphingomyelin content of the erythrocyte membranes. The negatively charged phosphate moiety of phospholipids of the membranes appears to inhibit a large amount of bilirubin binding to the membrane as its removal by phospholipase C greatly enhanced the binding. Furthermore, membrane proteins and carbohydrate also seem to play a significant regulatory function on the binding as their degradation and/or removal in the form of glycopeptides by trypsin expose a large number of bilirubin binding sites. PMID- 11423309 TI - Hemodynamics of local cerebral blood flow induced by somatosensory stimulation under normoxia and hyperoxia in rats. AB - We observed changes in the local cerebral blood flow (LCBF), red blood cell (RBC) concentration and RBC velocity in alpha-chloralose anesthetized rats using laser Doppler flowmetry during activation of the somatosensory cortex following electrical stimulation of the hind paw under hyperoxia (PaO(2)=513.5+/-48.4 mmHg; mean+/-S.D.) and normoxia (PaO(2)=106.4+/-8.4 mmHg). Electrical stimuli of 5 and 10 Hz (pulse width 0.1 ms) with an intensity of 1.5 mA were applied for 5 s (n=13 at 5 Hz, n=9 at 10 Hz). Baseline levels of LCBF and RBC concentration under hyperoxia were, respectively, 5.6+/-3.3 and 8.8+/-3.0% lower than those under normoxia (P<0.05), and that of RBC velocity under hyperoxia was slightly higher than that under normoxia (NS), suggesting mild vasoconstriction at rest under hyperoxia. At 5 Hz stimulation, after normalization to each baseline level, normalized response magnitudes of LCBF, RBC concentration and RBC velocity under hyperoxia were, respectively, 68.2+/-48.0, 71.1+/-65.5 and 66.0+/-56.3% greater than those under normoxia (P<0.05). At 10-Hz stimulation, normalized response magnitudes of LCBF and RBC concentration under hyperoxia were, respectively, 44.6+/-32.0 and 55.9+/-43.5% greater than those under normoxia (P<0.05), although a significant difference in the normalized response magnitude of RBC velocity was not detected between both conditions. The evoked LCBF under hyperoxia increased earlier, by approximately 0.15 s, than that under normoxia regardless of the stimulus frequency (P<0.05). These results suggest the involvement of oxygen interaction on the regulation of LCBF during neuronal activation. PMID- 11423310 TI - Variation of lipid classes among organs of the Northern krill Meganyctiphanes norvegica, with respect to reproduction. AB - Lipid content and class in the Northern krill Meganyctiphanes norvegica (digestive gland, stomach, gonad, fat body, abdomen) was investigated and correlated with sex and reproductive stage. Ready to spawn females, have high lipid content in ovaries, while in males and spent females, the major site of lipid deposits was the digestive gland, followed by the fat body. These differences among spawning and spent females are indicative of strong interactions between the ovary and digestive gland and the ovary and fat body during vitellogenesis. Triacylglycerols (TAG) were the major neutral lipid class with high levels in the digestive gland. The major phospholipid was phosphatidylcholine (PC) particularly in the muscular tissue of the abdomen. Phosphatidyl-ethanolamine (PE) and -serine-inositol (PS-PI), were present at intermediate levels. Reproductive males were depleted in TAG and diacylglycerols (DAG) in the digestive gland, gonad and fat body, and had 4 times lower cholesterol in the gonad than ready to spawn females. Furthermore, ready to spawn females had in the ovary higher amounts of TAG, DAG and phospholipids (PC, PE, PS PI) than spent females. Linear relationships between lipid content and main lipid class (TAG, PC, PE, PS-PI) in different fractions of males and ready to spawn females showed that: (1). TAG was stored for both sexes in all cephalothorax fractions with highest values in the digestive gland and ovary fluid; (2). PC was accumulated for both sexes in the fat body and the gonad with a higher slope for females, with the highest values in the ovary fluid and in the abdomen of males and that (3). PS-PI was stored only in the ovary and abdomen of mature females. These results are discussed in terms of the strategy developed by Meganyctiphanes norvegica to allocate lipids to the next generation for optimised embryogenesis. PMID- 11423311 TI - In vitro effect of temperature on phagocytic and cytotoxic activities of splenic phagocytes of the wall lizard, Hemidactylus flaviviridis. AB - The in vitro effect of temperature on phagocytosis, nitric oxide production and interleukin-1 (IL-1) secretion by splenic phagocytes isolated from the wall lizard (Hemidactylus flaviviridis) demonstrated that changes in temperature altered non-specific defenses. The LPS-induced percentage phagocytosis and phagocytic index were recorded maximum at 25 degrees C. The phagocytic activity declined considerably when the phagocytes were incubated at low (7 and 15 degrees C) or high (37 degrees C) temperatures. The presence of bacterial lipopolysaccharide (LPS) in the incubation medium could considerably enhance the phagocytic activity of splenic phagocytes. A similar temperature-related effect was also observed on LPS-induced cytotoxic activity of phagocytes. LPS could stimulate the nitrite release indicating nitric oxide production only at 25 degrees C. Likewise, the proliferative responses of immature rat's thymocytes to LPS-induced phagocyte-conditioned medium suggest that IL-1 secretion was enhanced when phagocytes were cultured at 25 degrees C. This suggests that 25 degrees C is the optimal temperature for phagocyte functions in H. flaviviridis. The decrease or increase in temperature other than at 25 degrees C dramatically suppressed the phagocyte activities. PMID- 11423312 TI - ACTH does not mediate divergent stress responsiveness in rainbow trout. AB - Two lines of rainbow trout selected for high (HR) and low (LR) responsiveness to a standardised confinement stressor displayed a sustained divergence in plasma cortisol levels during a 3-h period of confinement (max.: HR: 167+/-13 ng ml(-1); LR: 103+/-8 ng ml(-1); P<0.001). However, no significant difference in plasma ACTH levels was evident (max: HR: 153+/-9 pg ml(-1); LR: 142+/-7 pg ml(-1)). Dexamethasone (DEX) was administered to HR and LR fish to block endogenous adrenocorticotropin (ACTH) release. Administration of a weight-adjusted dose of ACTH to the DEX-blocked fish elevated plasma cortisol levels to a significantly greater extent in HR (233+/-24 ng ml(-1)) than LR (122+/-14 ng ml(-1)) fish (P<0.001). Plasma cortisol levels in DEX-blocked HR and LR fish after sham injection were low but also significantly different (HR: 6.7+/-1 ng ml(-1); LR: 2.2+/-0.2 ng ml(-1); P<0.001). These results indicate that modulation of cortisol responsiveness to stressors in HR and LR fish resides, at least in part, downstream of the hypothalamic-pituitary axis. PMID- 11423313 TI - Renal function in suckling and fasting pups of the northern elephant seal. AB - Elephant seals fast for prolonged periods without access to water. This is made possible, in part, by reductions in urine production. However, the mechanisms involved in reducing urine production are not understood. In this study, glomerular filtration rate (GFR) was measured in five northern elephant seal pups (Mirounga angustirostris) via the inulin clearance technique. Measurements were made during day 9 and day 18-22 of nursing and the second and eighth week of the postweaning fast. Plasma aldosterone and cortisol concentrations, quantified by radioimmunoassay, were measured in eight other weanlings during the second and eighth week of the fast. Mean GFR was 79.3+/-29.3 ml/min during the early suckling period and 78.2+/-17.1, 89.8+/-52.7, and 80.4+/-12.2 ml/min during the late suckling, early fasting and late fasting periods, respectively. Differences between nursing and fasting were insignificant, possibly because reduced protein oxidation during suckling and rapid recruitment of protein for tissue synthesis obviated the need for postprandial hyperfiltration. Alternatively, maintenance of GFR during fasting may facilitate urea concentration by compensating for reductions in the fractional excretion of urea. It is further hypothesized that aldosterone is primarily responsible for mediating renal water reabsorption in this system. PMID- 11423314 TI - Review: Protein function at thermal extremes: balancing stability and flexibility. AB - No organism can survive across the entire temperature range found in the biosphere, and a given species can rarely support active metabolism across more than a few tens of degrees C. Nevertheless, life can be maintained at surprisingly extreme temperatures, from below -50 to over 110 degrees C. That proteins, which are assembled with the same 20 amino acids in all species, can function well at both extremes of this range illustrates the plasticity available in the construction of these macromolecules. In studying proteins from extremophiles, researchers have found no new amino acids, covalent modifications or structural motifs that explain the ability of these molecules to function in such harsh environments. Rather, subtle redistributions of the same intramolecular interactions required for protein stabilization at moderate temperatures are sufficient to maintain structural integrity at hot or cold extremes. The key to protein function, whether in polar seas or hot springs, is the maintenance of an appropriate balance between molecular stability on the one hand and structural flexibility on the other. Stability is needed to ensure the appropriate geometry for ligand binding, as well as to avoid denaturation, while flexibility is necessary to allow catalysis at a metabolically appropriate rate. Comparisons of homologous proteins from organisms spanning a wide range of thermal habitats show that adaptive mutations, as well as stabilizing solutes, maintain a balance between these two attributes, regardless of the temperature at which the protein functions. PMID- 11423315 TI - Review: How was metazoan threshold crossed? The hypothetical Urmetazoa. AB - The origin of Metazoa remained--until recently--the most enigmatic of all phylogenetic problems. Sponges [Porifera] as "living fossils", positioned at the base of multicellular animals, have been used to answer basic questions in metazoan evolution by molecular biological techniques. During the last few years, cDNAs/genes coding for informative proteins have been isolated and characterized from sponges, especially from the marine demosponges Suberites domuncula and Geodia cydonium. The analyses of their deduced amino acid sequences allowed a molecular biological resolution of the monophyly of Metazoa. Molecules of the extracellular matrix/basal lamina, with the integrin receptor, fibronectin and galectin as prominent examples, cell-surface receptors (tyrosine kinase receptors), elements of nerve system/sensory cells (metabotropic glutamate receptor), homologs/modules of an immune system [immunoglobulin-like molecules, SRCR- and SCR-repeats, cytokines, (2-5)A synthetase], as well as morphogens (myotrophin) classify the Porifera as true Metazoa. As "living fossils", provided with simple, primordial molecules allowing cell-cell and cell-matrix adhesion, as well as processes of signal transduction as known in a more complex manner from higher Metazoa, sponges also show peculiarities. Tissues of sponges are rich in telomerase activity, suggesting a high plasticity in the determination of cell lineages. It is concluded that molecular biological studies with sponges as models will not only help to understand the evolution to the Metazoa but also the complex, hierarchical regulatory network of cells in higher Metazoa [reviewed in Progress in Molecular Subcellular Biology, vols. 19, 21 (1998) Springer Verlag]. The hypothetical ancestral animal, the Urmetazoa, from which the metazoan lineages diverged (more than 600 MYA), may have had the following characteristics: cell adhesion molecules with intracellular signal transduction pathways, morphogens/growth factors forming gradients, a functional immune system, and a primordial nerve cell/receptor system. PMID- 11423316 TI - The effects of thermal and hydric environments on hatching success, embryonic use of energy and hatchling traits in a colubrid snake, Elaphe carinata. AB - We examined the effects of thermal and hydric environments on hatching success, the embryonic use of energy and hatchling traits in a colubrid snake, Elaphe carinata. The eggs were incubated at four temperatures ranging from 24 to 32 degrees C on substrates with water potentials of 0 and -220 kPa using a 4x2 factorial design. Both thermal and hydric environments affected the water exchange between eggs and their surroundings. Eggs incubated in wetter substrates gained mass throughout the course of incubation, whereas eggs in drier substrates gained mass during the first half of incubation and lost mass thereafter. Hatching success was noticeably higher at 26 and 30 degrees C than at 24 and 32 degrees C, but among treatments, differences in hatching success were not significant. Temperature significantly affected the duration of incubation and most hatchling traits examined. Deformed hatchlings were found in all temperature treatments, with more deformities observed at 32 degrees C. Hatchlings from eggs incubated at different temperatures differed in wet body mass, but the differences stemmed mainly from variation in water contents. Embryos at different temperatures completed development at nearly the same expenditure of energy and catabolized nearly the same amount of lipids, but hatchlings from different temperatures differed in the development condition of carcass at hatching. Hatchlings from eggs incubated at 26 degrees C were larger in SVL than those from other higher or lower incubation temperatures, characteristically having larger carcasses; hatchlings from 32 degrees C eggs were smaller in SVL and had smaller carcasses but larger residual yolks than those from lower incubation temperatures. Hatchlings from eggs incubated at 24 degrees C were shorter in tail length but greater in size (SVL)-specific body wet mass than those from higher incubation temperatures. Within the range from -220 to 0 kPa, the substrate water potential did not affect hatching success, the embryonic use of energy and all hatchling traits examined, and the effects of temperature were independent of the effects of substrate water potential. Therefore, our data add evidence showing that embryonic development in reptiles with pliable-shelled eggs is relatively insensitive to variation in hydric environments during incubation. PMID- 11423317 TI - Effects of temperature and oxygen availability on circulating catecholamines in the toad Bufo marinus. AB - The release of catecholamines during hypoxia has received limited attention in amphibians and the adrenergic regulation of cardio-pulmonary functions is, therefore, not well understood at the organismic level. To describe the changes in plasma catecholamine concentrations, we exposed toads (Bufo marinus) to different levels of hypoxia at two temperatures (15 and 25 degrees C). In addition, blood oxygen binding properties were determined in vitro at 15 and 25 degrees C at two different pH values. Hypoxia elicited a significant increase in plasma catecholamines (adrenaline and noradrenaline) at both temperatures, in spite of a respiratory alkalosis. At 15 degrees C, the increase was from 2.6+/ 1.0 in normoxia to 4.8+/-1.4 ng ml(-1) at an inspired oxygen fraction of 0.05. At 25 degrees C, the hypoxic release of catecholamines was significantly higher (maximum levels of 44.8+/-11.6 ng ml(-1)). Plasma noradrenaline concentration was elevated at the most severe hypoxic levels, suggestive of an adrenal release. The arterial oxygen threshold for catecholamine release were approximately 1.0 mmol O(2) l(-1) blood or a PaO(2) of 30 mmHg. The P(50) values at 15 degrees C were 23.5+/-0.7 and 28.9+/-1.0 mmHg at pH 7.98+/-0.01 and 7.62+/-0.02, respectively, and increased to 36.5+/-0.6 and 43.0+/-1.1 mmHg at pH 8.04+/-0.04 and 7.67+/ 0.05, respectively, at 25 degrees C. The oxygen equilibrium curves were linear when transformed to Hill-plots and Hills n (the haemoglobin subunit co operativity) ranged between 2.24 and 2.75. The in vitro blood O(2) binding properties corresponded well with in vivo data. PMID- 11423318 TI - The influence of "host release factor" on carbon release by zooxanthellae isolated from fed and starved Aiptasia pallida (Verrill). AB - Symbiotic dinoflagellates (zooxanthellae) typically respond to extracts of host tissue with enhanced release of short-term photosynthetic products. We examined this "host release factor" (HRF) response using freshly isolated zooxanthellae of differing nutritional status. The nutritional status was manipulated by either feeding or starving the sea anemone Aiptasia pallida (Verrill). The release of fixed carbon from isolated zooxanthellae was measured using 14C in 30 min experiments. Zooxanthellae in filtered seawater alone released approximately 5% of photosynthate irrespective of host feeding history. When we used a 10-kDa ultrafiltrate of A. pallida host tissue as a source of HRF, approximately 14% of photosynthate was released to the medium. This increased to over 25% for zooxanthellae from anemones starved for 29 days or more. The cell-specific photosynthetic rate declined with starvation in these filtrate experiments, but the decline was offset by the increased percentage release. Indeed, the total amount of released photosynthate remained unchanged, or even increased, as zooxanthellae became more nutrient deficient. Similar trends were also observed when zooxanthellae from A. pallida were incubated in a 3-kDa ultrafiltrate of the coral Montastraea annularis, suggesting that HRF in the different filtrates operated in a similar manner. Our results support the suggestion that HRF diverts surplus carbon away from storage compounds to translocated compounds such as glycerol. PMID- 11423319 TI - Muscle fibre growth in undernourished reindeer calves (Rangifer tarandus tarandus L.) during winter. AB - To study whether moderate under-nutrition causes muscle wasting, reindeer (Rangifer tarandus tarandus L.) calves were fed either pelleted reindeer feed ad libitum (n=8) or restricted amounts of lichens (n=8). The restricted amount was 60% of ad libitum intake of lichens, and the feeding period was 6 weeks preceded by a 2-week adjustment period. Biopsy samples from the middle gluteal muscle (M. gluteus medius) for the analysis of fibre composition and area, as well as for the activity of cathepsin B were taken before the restriction period in November and January, and after the restriction period in April. In all calves the muscle fibre composition remained unchanged during the winter. In the lichen group, the fibre size also remained unchanged, whereas in control calves the cross sectional area of type I and type IIA fibres increased significantly from November to April. Cathepsin B activity decreased in all calves from November to January and remained at that low level for the rest of the study period, which suggests an attenuated rate of protein degradation. These results can be taken as an indication that moderate under-nutrition causes no muscle wasting in reindeer calves, and the decreased availability of nitrogen is partially compensated for by adaptive decrease in protein degradation. Interestingly the adaptive changes in protein metabolism are equally well seen in the well-fed controls as in the undernourished lichen-fed reindeer. PMID- 11423320 TI - Characterization and immunological analysis of ferritin from the hemolymph of Galleria mellonella. AB - Ferritin, an iron-binding protein, was purified from the larval hemolymph of the wax moth, Galleria mellonella by KBr density ultracentrifugation and FPLC (Superose 6). The iron content of ferritin was determined by atomic emission spectroscopy and Ferene S stain. Native molecular mass of ferritin was estimated as 630 kDa. SDS-PAGE revealed that the ferritin consists of two major polypeptides of 26 and 32 kDa and one minor polypeptide of 30 kDa. An isoelectric point of ferritin was measured to be approximately 7.3 and only the 32-kDa subunit is glycosylated. The ferritin contains large amounts of lysine, glutamine, glutamic acid and leucine but tryptophan was not detected. Electron microscopic examination of negatively stained preparations showed an 11-nm particle in external diameter and 7-nm iron core. Ferritin is present in both the ovary and testis. Localization of ferritin by immunoelectron microscopy in ovary and testis revealed that the gold particles were located in vitelline membrane and yolk granules but not in follicular epithelium of ovary. In the testis, the gold particles were located in testicular fluid and lumen of vas deferens. PMID- 11423321 TI - Hydrogen ion binding properties of tuna haemoglobins. AB - Tunas are very active fish with a high aerobic capacity, but they also regularly perform burst swimming with massive production of lactic acid. The present study examines whether H(+) buffering by tuna haemoglobin (Hb) is elevated to cope with metabolic acidoses (by analogy with the high buffer capacity of tuna white musculature) or whether the Hb-H(+) binding properties resemble other teleosts that have low buffer values and high Haldane effects. H(+) titration of oxygenated and deoxygenated composite Hb from yellowfin tuna, skipjack tuna and bigeye tuna in 0.1 M KCl revealed low Hb-specific buffer values in all three tunas. Values at physiological pH were comparable to those reported in less active species such as carp and eel. The fixed acid Haldane effect was large (maximal uptakes of close to 4 mol H(+) per mol Hb tetramer upon deoxygenation). Thus, the Hb-H(+) binding properties of very active tuna species correspond to other teleosts. Low Hb buffer values may be a pre-requisite for the regulation of red blood cell pH via Na(+)/H(+) exchange. Approximately nine "neutral" groups were titratable in tuna Hbs, suggesting that two alpha-amino groups and seven histidine residues are titrated within each tetramer. PMID- 11423322 TI - The thermal environment of arboreal pools and its effects on the metabolism of the arboreal, oophagous tadpoles of a Taiwanese tree frog, Chirixalus eiffingeri (Anura: Rhacophoridae). AB - We have studied seasonal and diurnal fluctuations of water temperature in bamboo stumps and the effect of temperature on the energy metabolism of arboreal, oophagous tadpoles of Chirixalus eiffingeri. We collected tadpoles (Gosner stage 28-29) in February and August from Chitou, Taiwan and acclimated them to 12 and 22 degrees C. Using a closed system, we measured tadpole oxygen consumption (V.O(2)) at 12, 17 and 22 degrees C. The water temperature was lowest in February (11-13 degrees C), increased rapidly during March and April and was highest from May to August (20-24 degrees C). Diel fluctuations in the temperature of the pools of water in bamboo stumps mirrored fluctuations in air temperature. Tadpoles collected in February and August exhibited metabolic compensation in that tadpoles acclimated at 12 degrees C had significantly higher V.O(2) than those acclimated at 22 degrees C. There are at least two possible explanations for the presence of metabolic compensation in C. eiffingeri tadpoles. Firstly, the larval period of C. eiffingeri ranges from 40 to 78 days, a tadpole could experience relatively large fluctuations in body temperature (up to 10 degrees C) during the development. As a result, C. eiffingeri tadpoles most likely evolved metabolic compensation to maintain activity levels under different thermal environments. Secondly, because arboreal pools are small, thermally unstratified, aquatic microhabitats, tadpoles are unable to behaviorally select preferred temperatures. As a result, metabolic compensation allows tadpoles to regulate their physiological functions. PMID- 11423323 TI - Effects of exercise on nitrogen excretion, carbamoyl phosphate synthetase III activity and related urea cycle enzymes in muscle and liver tissues of juvenile rainbow trout (Oncorhynchus mykiss). AB - The purpose of this study was to determine if carbamoyl phosphate synthetase III (CPSase III) and related urea cycle enzyme activities in skeletal muscle tissue of juvenile rainbow trout (Oncorhynchus mykiss) increase during short- or long term exercise, in parallel with changes in whole-body urea excretion rates. Urea excretion was elevated by 65% in fish that swam at high-speed (50 cm/s) vs. low speed (20 cm/s) over a 2-h period, with no significant changes in CPSase III, ornithine transcarbamoylase or glutamine synthetase activities in muscle tissue. Fish that swam for 4 days at high-speed had higher rates of ammonia excretion and GSase activity in muscle and liver tissue relative to low-speed swimmers. Calculations showed that 47-53% of excreted urea, theoretically could be accounted for by total muscle CPSase III activity in juvenile and adult trout. The data indicate that increases in the rate of urea excretion during short-term high intensity exercise are not linked to higher activities of urea cycle enzymes in muscle tissue, but this does not rule out the possibility of increased flux through muscle CPSase III and related enzymes. Furthermore, these results indicate that urea cycle enzyme activities in skeletal muscle tissue can account for a significant portion of total urea excretion in juvenile and adult trout. PMID- 11423324 TI - The effect of social context and reproductive status on the metabolic rates of dwarf seahorses (Hippocampus zosterae). AB - Although we are beginning to understand the conditions in which monogamy is favored over a more promiscuous lifestyle, little is known about the proximate effects of monogamous pair bonding, and subsequent reproduction, on general metabolism. I determined the effects of these factors on the metabolic rates of dwarf seahorses (Hippocampus zosterae), recognized for their monogamous lifestyle and unique male care of offspring, in a sealed brood pouch where embryos develop until birth. Resting routine oxygen consumption rates were measured in newly paired and reproductive adults using a continuous flow respirometer, and then compared to metabolic rates of sexually-isolated fish. Sex differences were observed in the relationship between log(10)mass and log(10)oxygen consumption, with pair-bonded females exhibiting a significantly higher slope than either pair bonded males, or sexually-isolated fish. Mass-specific metabolic rates in sexually-isolated fish were 15% higher than in pair-bonded fish, indicating that social conditions can strongly influence metabolic rate. Specific metabolic rates only differed by gender during male pregnancy, when male metabolic rate increased from 10 to 52% over pre-gravid levels. A male's developing brood only explained 4 31% of this increase, suggesting that increased metabolic demands on fathers accounts for most of the increase in metabolic rate during gestation. This study suggests that pair bonding can strongly affect the general metabolism of organisms, with potential differences between males and females that increase with age. PMID- 11423325 TI - An infrared thermographic study of surface temperature in the euthermic woodchuck (Marmota monax). AB - Surface temperatures were measured in euthermic woodchucks (Marmota monax) using infrared thermography across a range of ambient temperatures from -10 degrees C to 32 degrees C. The woodchuck keeps surface temperature of the peripalpebral region uniformly high, while head and body surfaces change proportionally with ambient temperature. When ambient temperature was below 0 degrees C, all surface temperatures increased which prevents freezing. At no point did the animals appear to be unable to regulate heat exchange. This species appears to be especially well adapted to the higher temperatures it encounters in its range. Vasomotion in the feet and to a lesser extent in the pinnae was used to regulate heat loss. At ambient temperature of 32 degrees C, mean temperatures of nose surfaces were 0.2 degrees C and 0.3 degrees C less than ambient temperature suggesting a type of counter current cooling mechanism may be present. PMID- 11423326 TI - Glucose-dependent insulinotropic polypeptide (GIP) and the enteroinsular axis in equines (Equus caballus). AB - To investigate the enteroinsular axis (EIA) in equines oral (oGTT) and intravenous (i.v.GTT) glucose tolerance tests (5.6 and 1 mmol glucose/kg BW, respectively) were performed with healthy, normal weight large horses and Shetland ponies. Plasma was analysed for concentrations of glucose, glucose dependent insulinotropic polypeptide (GIP) and insulin. In all equines plasma GIP concentrations only increased significantly when glucose was administered orally. The insulin glucose ratio (IGR) was significantly higher during the oGTT than during the i.v.GTT in both races. Basal plasma glucose levels were significantly higher in large horses than in ponies in both experiments. During the oGTT maximum glucose values were significantly higher in ponies. Ponies tended to a higher insulin secretion but the IGRs were identical in both races after oral and intravenous glucose administration. One clinically inconspicuous pony showed hyperinsulinaemia and, in case of the oGTT, insulin resistance, glucose intolerance, and GIP hypersecretion. The results of this study indicate the existence of an EIA in equines due to the higher IGRs during the oGTT. Furthermore, the similarity of plasma GIP levels and IGRs in ponies and large horses suggest a comparable activity of the EIA in both races. Regarding the elevated plasma GIP concentrations of the insulin resistant pony the EIA appears to participate in equine hyperinsulinaemia. PMID- 11423327 TI - Influence of alpha-linked glucose on jejunal sodium-glucose co-transport activity in ruminants. AB - Eight steers and 12 lambs were used in a completely randomized experimental design to determine the effect of partial alpha-amylase starch hydrolysate (SH) on small intestinal sodium-dependent glucose transport activity. Starch hydrolysate was delivered ruminally or abomasally to steers (960 g/day) and sheep (144 g/day) for 7 days. On day 7, the steers were rendered unconscious, exsanguinated and eviscerated. A 1-m section of jejunum was collected starting at the duodenojejunal flexure. Sheep were anaesthetized with pentobarbital and the second meter of small intestine (jejunum) was collected. Brush-border membrane vesicles were prepared and sodium-dependent glucose uptake activity was measured using the rapid uptake/filtration technique. Alkaline phosphatase and maltase activity was enriched by 8.2+/-0.5- and 8.4+/-1.2-fold in the vesicle preparation, respectively, and was not different between treatments. Abomasal SH increased (P=0.03) the Na/glucose co-transport approximately two-fold in both cattle (47.2-114.0+/-31.5 pmol/mgxsec) and sheep (77.4-152.0+/-25.7 pmol mg(-1) s(-1)). We conclude that Na/glucose co-transport activity by enterocytes responds to luminal alpha-linked glucose (from abomasal infusion) in ruminants, compared with controls. Intestinal maltase-specific activity does not respond to alpha linked glucose in cattle, and decreases slightly in sheep. PMID- 11423328 TI - Relationship of insulin-like growth factor-I and insulin to size and adiposity of under-yearling chinook salmon. AB - Sub-yearling spring chinook salmon were fed either a LoFat or HiFat diet from February to November. Fish were sampled over 2 days in November, following 24- and 48-h fasts. Length vs. weight relationships between fish fed the two diets were similar; however, fish fed the HiFat diet had roughly twice the body lipid as fish fed the LoFat diet (9% vs. 4.5%, respectively). Plasma IGF-I vs. length relations between fish fed the two diets were similar; overall, there was a strong relation between plasma IGF-I and length (r(2)=0.53). Similarly, plasma log (insulin) vs. length relations did not vary between the two diets; however, the relationship of log (insulin) vs. length was weak (r(2)=0.2). There was little or no relationship between plasma IGF-I or log (insulin) and body adiposity. Finally, there was a weak relationship between plasma IGF-I and log (insulin) (r(2)=0.23). PMID- 11423329 TI - Betaine aids in the osmoregulation of duodenal epithelium of broiler chicks, and affects the movement of water across the small intestinal epithelium in vitro. AB - In Experiment 1, the water holding capacity of broiler chick intestinal tissue was studied in vitro. The chicks were fed with corn-based diets with or without a 0.2% betaine supplementation in the drinking water. Slices from duodenum and jejunum were incubated in iso-osmotic (300 mM) or hyperosmotic saline (600 mM) with or without 10 mM betaine. The water volume of tissue slices was studied by adding tritiated water in the incubation medium while [14C]inulin was used to correct for the adherent water. After 30 min of incubation, by which time the steady-state of tritium influx had been achieved, the 3H and 14C-activities of the tissue slices were measured. The ileal and duodenal tissues incubated in the hyperosmotic saline accumulated less tritium than those incubated in iso-osmotic saline. Duodenal slices incubated in hyperosmotic saline with the presence of betaine showed a tritium content similar to slices incubated in iso-osmotic saline. The data suggest that the presence of betaine helped the duodenal, but not jejunal, epithelium to maintain water balance in hyperosmotic conditions. The dietary betaine supplementation diminished the differences between the incubation treatments in duodenal, but not in ileal tissue. In Experiment 2, the same double labeling method, but with shorter incubation times, was used to assess the rate of water flux from the incubation medium to duodenal or jejunal slices. The dietary treatments (as in Experiment 1) had little effect on the results. Betaine in the hyperosmotic saline significantly decreased the rate of tritium accumulation into the tissue slices, indicating that betaine slowed down the influx of water to the epithelium. We suggest that betaine affects the movement of water across the intestinal epithelium and has a role in the osmoregulation of small intestine of broiler chicks. PMID- 11423330 TI - Nitric oxide synthase activity in brain tissues from llama fetuses submitted to hypoxemia. AB - The fetal llama (Lama glama; a species adapted to live in chronic hypoxia in the highlands of the Andes) did not increase cerebral blood flow and reduce the brain oxygen uptake during hypoxemia. Although nitric oxide (NO) is a normal mediator in the regulation of vascular tone and synaptic transmission, NO overproduction by hypoxemia could produce neuronal damage. We hypothesized that nitric oxide synthase (NOS) activity is either maintained or reduced in the central nervous system of the llama fetuses submitted to chronic hypoxemia. Approximately 85% of the Ca(2+)-dependent NOS activity was soluble, at least 12% was associated with the mitochondrial fraction, and less than 5% remains associated with microsomes. To understand the role of NO in chronic hypoxemia, we determined the effect of 24 h hypoxemia on NOS activity in the central nervous system. No changes in activity or the subcellular distribution of NOS activity in brain tissues after hypoxemia were found. We proposed that the lack of changes in NOS activity in the llama under hypoxemia could be a cytoprotective mechanism inherent to the llama, against possible toxic effects of NO. PMID- 11423331 TI - Mannose-receptor-mediated clearance of lysosomal alpha-mannosidase in scavenger endothelium of cod endocardium. AB - Mannose-receptor-mediated clearance of circulating glycoproteins was studied in Atlantic cod (Gadus morhua). Distribution studies with radioiodinated and fluorescently labelled ligands showed that cod liver lysosomal alpha-mannosidase and yeast invertase were rapidly eliminated from blood via a mannose specific pathway in liver parenchymal cells and endocardial endothelial cells of atrium and ventricle. Asialo-orosomucoid, a galactose-terminated glycoprotein, was cleared by liver only. In vitro studies were performed with primary cultures of atrial-endocardial endothelial cells (AEC), incubated at 12 degrees C in a serum free medium. Cod AEC endocytosed mannose-terminated glycoproteins (125I-alpha mannosidase, 125I-invertase, 125I-mannan, 125I-ovalbumin and unlabelled lysosomal alpha-mannosidase), whereas 125I-asialo-orosomucoid was not recognised. Uptake of radiolabelled mannose-terminated ligands was inhibited 80-100% in the presence of excess amounts of mannan, invertase, D-mannose, L-fucose or EGTA. Our results suggest that the cod endocardial endothelial cells express a specific Ca(2+) dependent mannose receptor, analogous to the mannose receptor on mammalian macrophages and liver sinusoidal endothelial cells. PMID- 11423332 TI - Purification and characterization of multiple glutathione S-transferase isozymes from Chironomidae larvae. AB - Glutathione S-transferase (GST) has been implicated in the process of biotransformation of polycyclic aromatic hydrocarbons and of other organic pollutants by Chironomidae larvae. We have purified and characterized GST from cytosolic fractions of Chironomidae larvae. GST with an M(r) of 23 kDa has been purified to homogeneity from larvae by centrifugation, size exclusion chromatography on Sephadex G25, and glutathione affinity and anion exchange chromatography. The purified enzyme exhibited moderate activity towards 1,2 dichloro-4-nitrobenzene, 1-chloro-2,4-dinitrobenzene, 4-nitropyridine-N-oxide, p nitrobenzyl chloride, ethacrynic acid, and cumene hydroperoxide. The enzyme was homogeneous on gel isoelectric focusing and on SDS gel electrophoresis. Its isoelectric point was estimated to be 5.5. The enzyme had a maximum activity at approximately pH 8 and showed activity between 30 and 40 degrees C. It became inactive at higher temperature (>50 degrees C) for 5 min. The N-terminal sequence analysis of the amino acids shows a high % of conserved regions in the enzyme. The enzyme activity was comparable to levels of metabolism observed by animal GST involved in the detoxification of xenobiotics. PMID- 11423333 TI - Effects of estradiol-17beta treatment on in vitro and in vivo synthesis of two distinct vitellogenins in tilapia. AB - Two distinct vitellogenins (VTG) were purified from the blood of estradiol-17beta (E(2))-injected tilapia, Oreochromis mossambicus. Enzyme-linked immunosorbent assays (ELISA) of each VTG were developed to examine effects of E(2) treatment on induction of VTG synthesis in the primarily cultured tilapia hepatocytes. Two VTG molecules (VTG210 and VTG140) had apparent molecular masses of 370 and 220 kDa by gel filtration and 210 and 140 kDa by SDS-PAGE, respectively. Western blot analyses showed that antibodies raised against the purified VTG210 and VTG140 reacted only with each protein band. Furthermore, ELISA for each VTG was specific for target VTG. When E(2) was added into the media of primarily cultured tilapia hepatocytes, VTG210 and VTG140 were both detected from E(2) concentrations of 1x10(-7) M and 5x10(-7) M, respectively. Time course experiments showed that there was a difference in the detection time of VTG210 and VTG140 after the hormone treatment. Although the injection of different E(2) doses induced both VTGs in the plasma of male tilapia, the concentration of VTG210 was nearly five to eight times higher than that of VTG140. These results suggest that E(2) is a direct inducer of both VTGs in the tilapia hepatocytes in vitro and in vivo, and that there is difference in the hormone response in inducing the VTGs in the tilapia hepatocytes. PMID- 11423334 TI - The decrease in plasma melatonin at metamorphic climax in Rana catesbeiana (bullfrog) tadpoles is induced by thyroxine. AB - Melatonin decreases in the plasma of Rana catesbeiana (bullfrog) tadpoles at the climax of metamorphosis when the thyroxine (T(4)) level peaks. Since melatonin inhibited thyroid function in vitro, it would be of interest to determine if the decline in plasma melatonin permits greater thyroid hormone secretion, or if the increasing levels of T(4) cause the climactic decrease in plasma melatonin. The reciprocal effects of administering T(4) or melatonin just prior to metamorphic climax were examined in tadpoles kept at 22 degrees C on an 18L:6D cycle. If melatonin functions as a thyroid antagonist at later metamorphic stages, administration of melatonin should decrease plasma T(4), whereas if T(4) causes the decline in plasma melatonin, T(4) treatment of tadpoles prior to climax should induce the climactic melatonin decrease prematurely. Once daily injection of 40 microg melatonin for 5 days at 19.30 h had no effect on metamorphic progress, or on plasma T(4) or melatonin levels, except for a transient rise in melatonin just after the injection. Immersion in 2.2x10(-4) M melatonin for 6 days accelerated metamorphosis and decreased plasma melatonin, but had no effect on plasma T(4). Administration of T(4) by injection of 0.2 microg, or immersion in a 6.3x10(-8) M solution accelerated metamorphosis more than melatonin immersion, raised plasma T(4) to climax levels, and induced a decrease in plasma melatonin. We conclude that rapid clearance of exogenous melatonin from the circulation in these experiments did not allow it to affect plasma T(4), and that there is clear evidence that the rise in T(4) induces the climax decrease in plasma melatonin. The finding that immersion in a high level of melatonin can lower plasma melatonin and accelerate metamorphosis, whereas a single daily injection does not, provides an explanation for some of the contradictory reports in the literature concerning melatonin's effect on tadpole metamorphic progress. PMID- 11423335 TI - Modulation of leucine absorption in the larval midgut of Bombyx mori (Lepidoptera, Bombycidae). AB - In the larval midgut of Bombyx mori a K(+)-dependent transporter for leucine and amino acids with a hydrophobic side chain is responsible for the absorption of most essential amino acids. We investigated if a modulation of its activity occurred as a result of starvation or after hormonal treatments. We measured amino acid uptake in brush border membrane vesicles (BBMV) purified from the anterior-middle (AM) and posterior (P) regions of the midgut in fifth instar larvae. Silkworms were either starved or topically treated with low dosages of fenoxycarb, a molecule often used as a juvenile hormone mimic. The maximal uptake value of K(+)-driven leucine transport was increased in BBMV of AM- and P-midgut regions of starved larvae. The initial uptake rates of serine and glutamine, two amino acids transported by the same cotransporter as leucine, were also increased. Leucine kinetics proved that V(max) was the kinetic parameter modified by starvation in both midgut regions. Topical applications of fenoxycarb at a dose of 2.5 fg/larva immediately after the fourth ecdysis, induced an increase of leucine initial uptake rates and of intravesicular accumulation of leucine in both AM- and P-BBMV. Kinetic analysis of leucine uptake indicated again that V(max) was increased in BBMV from both midgut regions in treated larvae. PMID- 11423336 TI - Effect of feeding on circulating micronutrient concentrations in the Burmese python (Python molurus). AB - Burmese pythons (Python molurus) regulate digestive performance and metabolism with the ingestion of each meal. To explore the python's postprandial responses, we monitored the concentrations of blood micronutrients and homocysteine during fasting and for 15 days after feeding. Plasma folate concentrations peaked with a 270% increase over fasting levels 3 days after feeding, whereas plasma B-12 peaked with a 66% increase within 1 day. Erythrocyte folate concentrations were highest 15 days after feeding with a 44% increase. The major plasma folate was 5 methyltetrahydrofolate during fasting and was non-5-methyltetrahydrofolate during digestion, whereas erythrocytes contained polyglutamyl forms of non-5 methyltetrahydrofolate. Plasma homocysteine concentrations peaked with a 56% increase 3 days after feeding, and were markedly greater than those of mammals. Plasma zinc and copper did not change significantly. Plasma zinc concentrations were 20 times greater than plasma copper and approximately 30 times higher than those of mammals. Pythons showed a significant postprandial decline of 25% in hematocrit. Plasma pyridoxal 5'-phosphate (coenzyme form of vitamin B-6) was not detected probably due to its tight protein binding. Most micronutrient concentrations appear to plateau 3 days after feeding, suggesting that pythons have relatively rapid homeostasis of micronutrients despite the ingestion of large meals. PMID- 11423337 TI - Cyclic CO(2) release in Cryptotermes cavifrons Banks, Incisitermes tabogae (Snyder) and I. minor (Hagen) (Isoptera: Kalotermitidae). AB - CO(2) release patterns of three drywood termite species were investigated using flow-through respirometry techniques. Eight hours of real-time CO(2) release data were recorded for pseudergates of Cryptotermes cavifrons Banks, Incisitermes minor (Hagen), and I. tabogae (Snyder) at 20-40 degrees C. Cyclic release of CO(2) was observed in 20-90% of C. cavifrons, 70-100% of I. tabogae, and 87-100% of I. minor pseudergates. Variability of the recordings (calculated as the coefficient of variability or CV) was used to estimate the level of cycling in each recording. CV ranged from 14.53+/-2.57 (40 degrees C) to 32.33+/-1.12% (30 degrees C) in C. cavifrons, 20.24+/-2.44 (35 degrees C) to 67.3+/-10.3% (20 degrees C) in I. minor, and 15.9+/-1.46 (35 degrees C) to 34.15+/-6.18% (20 degrees C) in I. tabogae. The relationship between temperature and CV for each species was modeled using non-linear regression. CV of both Incisitermes spp. decreased exponentially with temperature, while C. cavifrons CV followed a Gaussian model, indicating an optimal cycling temperature of approximately 30 degrees C. Mean V.CO(2) values were determined for each species as a function of temperature, and ranged from 0.1 ml CO(2) g(-1) h(-1) (I. minor at 20 degrees C) to 0.8 ml CO(2) g(-1) h(-1) (C. cavifrons at 40 degrees C). For all three species, V.CO(2) significantly increased linearly with temperature. Colinearity tests indicated that different models described the V.CO(2) relationship with temperature for both genera. Q(10) values for V.CO(2) over the range of 20-40 degrees C were 1.92 for I. minor, 1.66 for I. tabogae, and 1.62 for C. cavifrons pseudergates. PMID- 11423338 TI - Review: Analysis of the evolutionary convergence for high performance swimming in lamnid sharks and tunas. AB - Elasmobranchs and bony fishes have evolved independently for more than 400 million years. However, two Recent groups, the lamnid sharks (Family Lamnidae) and tunas (Family Scombridae), display remarkable similarities in features related to swimming performance. Traits separating these two groups from other fishes include a higher degree of body streamlining, a shift in the position of the aerobic, red, locomotor muscle that powers sustained swimming to a more anterior location in the body and nearer to the vertebral column, the capacity to conserve metabolic heat (i.e. regional endothermy), an increased gill surface area with a decreased blood-water barrier thickness, a higher maximum blood oxygen carrying capacity, and greater muscle aerobic and anaerobic enzyme activities at in vivo temperatures. The suite of morphological, physiological, and biochemical specializations that define "high-performance fishes" have been extensively characterized in the tunas. This review examines the convergent features of lamnid sharks and tunas in order to gain insight into the extent that comparable environmental selection pressures have led to the independent origin of similar suites of functional characteristics in these two distinctly different taxa. We propose that, despite differences between teleost and elasmobranch fishes, lamnid sharks and tunas have evolved morphological and physiological specializations that enhance their swimming performance relative to other sharks and most other high performance pelagic fishes. PMID- 11423339 TI - Red muscle function during steady swimming in brook trout, Salvelinus fontinalis. AB - Red muscle function during steady swimming in brook trout was studied through both in vivo swimming and in vitro muscle mechanics experiments. In the swimming experiments, red muscle activity was characterized through the use of electromyography and sonomicrometry, allowing the determination of several parameters such as tailbeat frequency, EMG burst duration, muscle length change patterns and relative phase of EMG activity and length change. Brook trout do show some shifts in these variables along their length during steady swimming, but the magnitude of these shifts is relatively small. In the muscle mechanics experiments, the in vivo muscle activity data were used to evaluate patterns of power production by red muscle during swimming. Unlike many fish species, the red muscle along the length of brook trout shows little change in isometric kinetic variables such as relaxation rate and twitch time. Furthermore, there is no rostral-caudal shift in red muscle mass-specific power output during steady swimming. This last result contrasts sharply with rainbow trout and with a variety of other fish species that power steady swimming primarily with the posterior red myotome. PMID- 11423340 TI - Genotoxic effect of 2,4-dichlorophenoxy acetic acid and its metabolite 2,4 dichlorophenol in mouse. AB - The cytogenetic effect of 2,4-dichlorophenoxy acetic acid (2,4-D) and its metabolite 2,4-dichlorophenol (2,4-DCP) was studied in bone-marrow, germ cells and sperm head abnormalities in the treated mice. Swiss mice were treated orally by gavage with 2,4-D at 1.7, 3.3 and 33 mg kg(-1)BW (1/200, 1/100 and 1/10 of LD(50)). 2,4-DCP was intraperitoneally (i.p.) injected at 36, 72 and 180 mg kg( 1)BW (1/10, 1/5, 1/2 of LD(50)). A significant increase in the percentage of chromosome aberrations in bone-marrow and spermatocyte cells was observed after oral administration of 2,4-D at 3.3 mg kg(-1)BW for three and five consecutive days. This percentage increased and reached 10.8+/-0.87 (P<0.01) in bone-marrow and 9.8+/-0.45 (P<0.01) in spermatocyte cells after oral administration of 2,4-D at 33 mg kg(-1)BW for 24 h. This percentage was, however, lower than that induced in bone-marrow and spermatocyte cells by mitomycin C (positive control). 2,4-D induced a dose-dependent increase in the percentage of sperm head abnormalities. The genotoxic effect of 2,4-DCP is weaker than that of 2,4-D, as indicated by the lower percentage of the induced chromosome aberrations (in bone-marrow and spermatocyte cells) and sperm head abnormalities. Only the highest tested concentration of 2,4-DCP (180 mg kg(-1)BW, 1/2 LD(50)) induced a significant percentage of chromosome aberrations and sperm head abnormalities after i.p. injection. The obtained results indicate that 2,4-D is genotoxic in mice in vivo under the conditions tested. Hence, more care should be given to the application of 2,4-D on edible crops since repeated uses may underlie a health hazard. PMID- 11423341 TI - Evaluation of genotoxic potential of synthetic progestins-norethindrone and norgestrel in human lymphocytes in vitro. AB - The genotoxicity study of two widely used contraceptive synthetic progestins, i.e. norgestrel and norethindrone was carried out on human lymphocyte chromosomes using chromosomal aberrations (CA), sister chromatid exchanges (SCE) and cell growth kinetics as parameters. The study was carried out both in the presence as well as in the absence of metabolic activation (S(9) mix). The lymphocytes were exposed to three different concentrations of the drugs (20, 40 and 75 microg/ml for norethindrone and 10, 25 and 50 microg/ml for norgestrel) for three different durations (24, 48 and 72 h). The drug norethindrone was found to be non-genotoxic at any concentration and at any exposure duration either in the presence or in the absence of S(9) mix. But another drug norgestrel was found to affect the genetic material. It induces CA, SCE at significant level, and inhibits lymphocyte proliferation at 25 and 50 microg/ml of concentrations only. In the presence of S(9) mix the values obtained for CA, SCE and mitotic index (MI) were more significant. A time and dose relationship was also observed. It was concluded that norgestrel itself and possibly its metabolites are potent mutagens beyond a particular dose in human lymphocytes. PMID- 11423342 TI - The effect of tobacco smoke, nicotine, and cotinine on the mutagenicity of 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL). AB - 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) is a rodent carcinogen that is metabolically derived from carbonyl reduction of 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone (NNK). NNAL can be pyridine N-oxidized to form NNAL-N-oxide, or conjugated to form NNAL-glucuronide - non-genotoxic metabolites that can be excreted in urine. Alternatively, NNAL can be alpha-hydroxylated at the methyl and methylene carbons adjacent to the nitroso group to generate electrophiles that can react with biological macromolecules, such as DNA and proteins. Our laboratory has previously demonstrated that the mutagenicity of NNK was significantly inhibited by the aqueous extract of tobacco smoke, as well as pyridine alkaloids in cigarette smoke, such as nicotine, cotinine and nornicotine. Given the structural similarity between NNK and NNAL, and the metabolic activation of both by cytochromes P450, we hypothesized that there may be a similar inhibition of NNAL metabolism, and consequently, inhibition of the mutagenic activity of NNAL by tobacco smoke and its pyridine alkaloid constituents. In the present study, we evaluated the ability of two pyridine alkaloids (nicotine and cotinine) and aqueous cigarette smoke condensate extract (ACTE) to inhibit the mutagenicity of NNAL in Salmonella typhimurium strain TA1535 in the presence of a metabolic activation system (S9). Both pyridine alkaloids tested, as well as ACTE, inhibited the mutagenicity of NNAL in a concentration-dependent manner. The observed reductions in mutagenicity were not the result of cell killing due to cytotoxicity. These results demonstrate that tobacco smoke contains pyridine alkaloids, as well as other unidentified constituents that inhibit the mutagenicity of NNAL, a major metabolite of NNK. PMID- 11423343 TI - Acetoxy-4-methylcoumarins confer differential protection from aflatoxin B(1) induced micronuclei and apoptosis in lung and bone marrow cells. AB - The ability of various acetoxy derivatives of 4-methylcoumarins to inhibit the genotoxic changes due to aflatoxin B(1) (AFB(1)) is reported here. Several 4 methylcoumarins (test compounds), such as 7,8-diacetoxy-4-methylcoumarin (DAMC), monoacetoxy-4-methylcoumarin (MAC), 5-N-acetyl-6-acetoxy-4-methylcoumarin (NAMC) and 7,8-dihydroxy-4-methylcoumarin (DHMC) were separately administered intraperitoneally (i.p.) to male wistar rats followed by AFB(1) administration i.p. or intratracheally (i.t.) (2-8 mg/kg b.wt.) and another dose of the test compound. The animals were sacrificed 26h after AFB(1) administration. From animals receiving AFB(1) i.p., bone marrow (BM) cells were isolated and stained with Mayer's haematoxylin and eosin. Micronuclei (MN) in BM were scored by light microscopy. From animals receiving AFB(1) i.t., bronchoalveolar lavage (BAL) was obtained, lung cells (LG) were isolated and stained with fluorochrome 6-diamidino 2-phenylindole (DAPI) for the analysis of MN, apoptotic bodies (AP) and cell cycle variations. Rats were separately treated with the vehicle DMSO to serve as the proper control. AFB(1) caused significant dose-dependent induction of MN in BM as well as LG. AP were observed in LG of rats receiving AFB(1) and was found to correlate with MN induction. DAMC injection caused significant decrease in AP due to AFB(1) in LG and MN in both BM and LG. The effectiveness of MAC was approximately half that of DAMC, thereby indicating that number of acetoxy groups on the coumarin molecule determine the efficacy. The fact that NAMC had no effect either on MN or AP indicate that neither acetoxy group at C-6 nor the N-acetyl group at C-5 facilitate the transfer of acetyl group to P-450 required for inhibition of AFB(1)-epoxidation. DHMC, the deacetylated product of DAMC had no normalizing effect on the induction of MN and AP. These findings confirm our earlier hypothesis that DAMC-mediated acetylation of microsomal P-450 (catalysing epoxidation of AFB(1)) through the action of microsomal transacetylase is responsible for the protective action of DAMC. The relative number and position of acetoxy groups on the coumarin nucleus determine the specificity to the transacetylase necessary for the chemopreventive action. PMID- 11423344 TI - Re-evaluation of the mutagenic potential of quinacrine dihydrochloride dihydrate. AB - Quinacrine has been used for voluntary female non-surgical sterilization for its ability to produce tubal occlusion. Safety issues regarding quinacrine have been raised because it has been shown to intercalate with DNA. Therefore, safety issues need to be resolved by appropriate toxicology studies to support a review for human transcervical use. Such toxicology studies include mutagenicity assays. Here we report an evaluation of the genotoxicity of quinacrine dihydrochloride dihydrate (QH) using a battery of assays. In the bacterial mutagenicity assay, QH was strongly positive in Salmonella typhimurium tester strain TA1537 with and without S9-activation and in S. typhimurium tester strain TA98 with S9 activation; QH was also strongly positive in Escherichia coli WP2 uvrA without S9 activation. QH was not mutagenic in S. typhimurium tester strains TA100 and TA1535 with and without S9-activation. QH was mutagenic in the mouse lymphoma assay in the absence of S9-activation. QH was clastogenic in Chinese hamster ovary (CHO) cells, with and without S9-activation. QH was negative for polyploidy in the same chromosome aberration test. Using a triple intraperitoneal injection treatment protocol in both male and female mice, QH was negative in the in vivo mouse micronucleated erythrocyte (micronucleus) assay. These results confirm that QH is mutagenic and clastogenic in vitro and suggest a potential risk to human health due to QH exposure after intrauterine exposure. PMID- 11423345 TI - In isolated DNA, formamidopyrimidine-DNA glycosylase-sensitive sites determined by electrophoresis correspond to the amount of 8-oxo-7,8-dihydro-2' deoxyguanosine by HPLC-ECD. AB - Several methods have been developed for determining the amount of 8-oxo-7,8 dihydro-2'-deoxyguanosine (8-oxodG) in DNA. In the present study, we compared an electrophoretic method that uses formamidopyrimidine-DNA glycosylase (FPG protein) with a HPLC-ECD method. Firstly, we produced 8-oxodG in lambda DNA with methylene blue and visible light and cleaved it in one-half of the modified DNA enzymatically with FPG protein. Then, we determined the number of FPG protein sensitive sites by electrophoresis (Y) and the number of 8-oxodGs by HPLC-ECD (X) per 10(5)dG of isolated DNA. Simple regression analysis of the data showed Y=1.07X+1.52 to be the most likely relationship. The correlation coefficient was 0.97. The values obtained by the two methods were very similar. This result is noteworthy because the number of FPG protein-sensitive sites determined by other methods have not yet come close to the number obtained by HPLC-ECD. Thus, this method might be more quantitative than other methods that measure FPG protein sensitive sites. Another reason this electrophoresis method might be more useful than HPLC-ECD is that we can determine some other types of oxidative DNA damage well, by changing the DNA glycosylase. PMID- 11423346 TI - Oxidative DNA damage by an N-hydroxy metabolite of the mutagenic compound formed from norharman and aniline. AB - Norharman (9H-pyrido[3,4-b]indole), which is a heterocyclic amine included in cigarette smoke or cooked foodstuffs, is not mutagenic itself. However, norharman reacts with non-mutagenic aniline to form mutagenic aminophenylnorharman (APNH), of which DNA adducts formation and hepatocarcinogenic potential are pointed out. We investigated whether N-OH-APNH, an N-hydroxy metabolite of APNH, can cause oxidative DNA damage or not, using 32P-labeled DNA fragments. N-OH-APNH caused Cu(II)-mediated DNA damage. When an endogenous reductant, beta-nicotinamide adenine dinucleotide (NADH) was added, the DNA damage was greatly enhanced. Catalase and a Cu(I)-specific chelator inhibited DNA damage, suggesting the involvement of H(2)O(2) and Cu(I). Typical -*OH scavenger did not inhibit DNA damage. These results suggest that the main reactive species are probably copper hydroperoxo complexes with DNA. We also measured 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodG) formation by N-OH-APNH in the presence of Cu(II), using an electrochemical detector coupled to a high-pressure liquid chromatograph. Addition of NADH greatly enhanced 8-oxodG formation. UV-VIS spectra and mass spectra suggested that N-OH-APNH was autoxidized to nitrosophenylnorharman (NO PNH). We speculated that NO-PNH was reduced by NADH. Cu(II) facilitated the redox cycle. In the presence of NADH and Cu(II), very low concentrations of N-OH-APNH could induce DNA damage via redox reactions. We conclude that oxidative DNA damage, in addition to DNA adduct formation, may play an important role in the expression of genotoxicity of APNH. PMID- 11423347 TI - Identification of genotoxic stress in human cells by fluorescent monitoring of p53 expression. AB - The tumor suppressor protein p53 is induced upon DNA damage essentially by post translational regulatory mechanisms, which lead to a substantial increase of p53 levels. To exploit this essential property of p53, we developed a novel reporter system for monitoring accumulation and subcellular translocation of p53 protein, which is able to function as a simple test for detecting mutagenic and genotoxic stress in human cells. For this purpose, we constructed a plasmid with a specific translational TP53::EGFP gene fusion and selected stable transfected clones in the human cell line HEK293, in which p53 is functionally stabilized due to the expression of the transgenic adenoviral E1A oncoproteins. HEK293-TP53::EGFP clones may be used as a living cell system for monitoring not only of the induction of p53 protein in the cell, but also of its subcellular localization. Using this human reporter cell system, we examined levels of p53 by fluorescence microscopy and by FACS analysis following treatment with several classes of genotoxic and carcinogenic compounds. All tested DNA damaging agents caused a significant increase of intracellular p53-EGFP levels in a concentration dependent manner. On the other hand, non-genotoxic carcinogens and stress conditions that cannot damage DNA were not able to induce p53-EGFP accumulation. The induction effect caused by genotoxic stress was found to be dependent on the endogenous p53 status, because it was not observed in p53-deficient cell lines. This corroborates the notion that p53 may be used as an universal sensor for genotoxic stress and demonstrates the usefulness of HEK293-p53-EGFP cells as a reporter system for identification of mutagens and genotoxic carcinogens in human cells by means of visualizing and monitoring intracellular p53 levels and localization. PMID- 11423348 TI - Frequency of HPRT mutants in humans exposed to vinyl chloride via an environmental accident. AB - The mutant frequency (MF) in the hypoxanthine-guanine-phosphoribosyl-transferase (HPRT) locus of peripheral blood T-lymphocytes was measured in a population environmentally exposed to vinyl chloride - a toxic and carcinogenic substance through an accidental release into the atmosphere. It was compared to MF in a control group of unexposed individuals. Both groups were re-investigated in a follow-up study, 2 years later. No significant difference could be observed in MF between exposed and controls either at the accident nor in the follow-up study. Approximately the same mean HPRT mutant frequencies were observed for both groups in T-lymphocytes from blood samples obtained shortly after the accident and from the follow-up blood samples. Both groups showed a higher mean MF in the re investigation samples which is most probably due to the significantly lower average cloning efficiency (CE) under non-selective conditions and because of the inverse relationship between CE and MF. The exposed population showed a higher mean T-cell CE at the initial blood sampling as compared to the control group. The concurrent cytogenetic analyses of peripheral lymphocytes showed a significant increase in cells with aberrations in the exposed population. Clastogenic but not mutagenic activity of vinyl chloride was observed in our study. PMID- 11423349 TI - Formation of DNA adducts in the aorta of smoke-exposed rats, and modulation by chemopreventive agents. AB - Our previous studies showed that nucleotide alterations, evaluated by (32)P postlabeling, are systematically detected in smooth muscle cells of atherosclerotic lesions localized in the aorta of surgical patients. The level of these molecular lesions was correlated with the occurrence of known atherogenic risk factors, among which the number of currently smoked cigarettes, and was significantly enhanced in individuals having a null GSTM1 genotype as compared to individuals carrying the GSTM1 genotype. The present study had the dual objective of evaluating the formation of DNA adducts in the whole thoracic aorta of Sprague Dawley rats, exposed whole-body to cigarette smoke for 28 consecutive days, and of investigating the effects of chemopreventive agents given orally during the same period. High levels of (32)P postlabeled DNA adducts were formed in the aorta of smoke-exposed rats, with an overall 11 times increase over the total levels observed in sham-exposed rats, and with increases ranging between three and 63 times for seven individual DNA adducts. Supplement of the diet with either 1,2-dithiole-3-thione, phenethyl isothiocyanate or 5,6-benzoflavone had no or poor effects on the smoke-related formation of nucleotide alterations in the aorta. In contrast, oltipraz, given with the diet, N-acetyl-L-cysteine, given with drinking water and, even more potently, their combination exerted remarkable protective effects. The results of this experimental study, together with the previous findings in humans, suggest that DNA alterations may contribute to the atherogenic process, clarify a possible mechanism of cigarette smoke, a well known atherogen, and show the potential protective effects of certain drugs towards these alterations. PMID- 11423350 TI - Resveratrol, a naturally occurring polyphenol, induces sister chromatid exchanges in a Chinese hamster lung (CHL) cell line. AB - We tested the genotoxicity of 3,5,4'-trihydroxystilbene (resveratrol), a polyphenolic phytoalexin found in grapes, in a bacterial reverse mutation assay, in vitro chromosome aberration (CA) test, in vitro micronucleus (MN) test, and sister chromatid exchange (SCE) test. Resveratrol was negative in the strains we used in the bacterial reverse mutation assay (S. typhimurium TA98 and TA100 and E. coli WP2uvrA) in the absence and presence of a microsomal metabolizing system. It induced structural CAs at 2.5-20 microg/ml and showed weak aneuploidy induction in a Chinese hamster lung (CHL) cell line. It induced MN cells and polynuclear and karyorrhectic cells after 48h treatments in the in vitro MN test. In the SCE test, resveratrol caused a clear cell-cycle delay; at 10 microg/ml, the cell cycle took twice as long as it did in the control. Resveratrol induced SCEs dose-dependently at up to 10 microg/ml, at which it increased SCE six-fold, and the number was almost as large as mitomycin C, a strong SCE inducer. No second mitoses were observed at 20 microg/ml even after 54h. Cell cycle analysis by FACScan indicated that resveratrol caused S phase arrest, and 48h treatment induced apoptosis. Our results suggest that resveratrol may preferentially induce SCE but not CA, that is, it may cause S phase arrest only when SCEs are induced. PMID- 11423351 TI - Mutagenicity of electrophilic N-acyloxy-N-alkoxyamides. AB - N-acyloxy-N-alkoxybenzamides are mutagenic in TA100 without the need for metabolic activation with S9. Electronic effects of substituents on both the benzamide ring in N-acetoxy-N-butoxybenzamides or the benzyloxy ring in N-acetoxy N-benzyloxybenzamides do not influence mutagenicity levels. For N-benzoyloxy-N benzyloxybenzamides, mutagenicity levels are inversely related to the electron withdrawing effect of substituents on the benzoyloxy leaving group. Since reactivities increase with increasing electron-withdrawing effects, mutagenicity correlates with stability rather than reactivity of these mutagens. Hydrophobicity is the dominant factor controlling mutagenicity levels and data for all mutagens correlate with computed logP values with a lower dependence (h=0.22) than that recorded for indirect mutagens (h=1.0), except where a sterically demanding p-tert-butyl substituent or a naphthyl group is present. N acetoxy-N-butoxynaphthamide exhibits a much higher level of mutagenicity than predicted by its logP value and activity may be ascribed to an intercalative binding process with DNA rather than straightforward hydrophobic binding in the major or minor groove. Since these are direct-acting mutagens, structural factors influence binding and reactivity towards DNA. PMID- 11423352 TI - The protection of Vitamin E and selenium against carbon tetrachloride-induced genotoxicity in ovine peripheral blood lymphocytes. AB - The protective effect of Vitamin E and selenium was studied for the possibility of decreasing the chromosome aberrations (CA), micronuclei (MN) and sister chromatid exchanges (SCE) induced by carbon tetrachloride (CCl(4)) in ovine peripheral lymphocytes cultured in vitro. The cultures of lymphocytes from two healthy lambs were treated with carbon tetrachloride at concentrations of 2, 4, 8, and 16 microg/ml for the last 48 h of cultivation and subsequently with the same doses of CCl(4) and Vitamin E and selenium. A possible metabolic modification in carbon tetrachloride genotoxicity was detected with the application of S9 fraction for 2h. No positive clastogenic effect of CCl(4), and subsequently no protective effect of either antioxidant was obtained in the CA assay. In the MN assay for 48 h, an increase and, respectively, a decrease in the frequency of MN was found in cultures treated with CCl(4) alone and in cultures treated concurrently with CCl(4), Vitamin E and selenium at concentrations of 8 and 16 microg/ml (P<0.001 and P<0.01). High statistical significance was achieved in SCE assay after the treatment of ovine peripheral lymphocytes with CCl(4) at all concentrations tested (P<0.001), except the lowest dose. A significant decrease in SCEs was obtained in cultures treated simultaneously with CCl(4), Vitamin E and selenium (P<0.001). The highest dose (16 microg/ml) was also shown to improve the cell-cycle kinetics in comparison with corresponding unprotected dose (P<0.01). Insufficient confirmation of the genotoxicity of CCl(4) and the protective effect of the antioxidants, respectively were seen in cultures with metabolic activation. PMID- 11423353 TI - Analyses of DNA adducts formed by ochratoxin A and aristolochic acid in patients with Chinese herbs nephropathy. AB - Chinese herbs nephropathy (CHN), a unique type of nephropathy has been associated with the intake of weight-reducing pills containing the Chinese herb Aristolochia fangchi. Moreover, an association between the use of A. fangchi and urothelial cancer in CHN patients has been reported indicating that aristolochic acid (AA) the major alkaloid of A. fangchi might be the causal agent. Similarities of CHN to the Balkan endemic nephropathy (BEN) have led to the hypothesis of a common etiological agent for both diseases. Evidence has accumulated that BEN is an environmentally-induced disease strongly associated with the fungal mycotoxin ochratoxin A (OTA). Both, AA and OTA are nephrotoxic and carcinogenic and induce the formation of DNA adducts. As OTA has been suspected as fungal contaminant in the herbal batches used for the preparation of the weight-reducing pills we analysed tissues from CHN patients by the 32P-postlabeling procedure for the presence of DNA adducts related to both OTA and AA exposure. Whereas, AA-specific DNA adducts were detected in all five urinary tract tissues from five patients (total RAL: 32-251 adducts per 10(9) nucleotides), OTA-related DNA adducts were detectable in two kidneys and one ureter only (total RAL: 1.5-3.7 adducts per 10(9) nucleotides). Thus, OTA-related DNA adduct levels were about 50 times lower than AA-DNA adduct levels. In female and male rats that were treated with the slimming regimen in the same way like the CHN patients except that the amount of Chinese herbs was 10 times higher, AA-DNA adducts were found in kidney tissues (total RAL ranging from 51 to 83 adducts per 10(9) nucleotides) but adducts derived from OTA were not observed. These results demonstrate that OTA-related DNA adducts do not play a key role in CHN or CHN-associated urothelial cancer. PMID- 11423354 TI - Micronuclei induced in erythrocytes of Cyprinus carpio (teleostei, pisces) by X rays and colchicine. AB - In the present work the induction of micronuclei in erythrocytes of Cyprinus carpio treated with X-rays and colchicine is studied for the evaluation of mutagenic effects of both clastogenic and mitoclastic (spindle poisoning) agents in this system. Three different experiments were performed treating groups of laboratory-reproduced animals with (1) single doses of X-rays (0.1, 0.5 and 2Gy); (2) a single i.p. injection of colchicine at the concentrations: 1.6x10(-2), 8x10(-2), 0.4 and 2mg/kg b.w. so as to mimic an acute exposure to the agent and (3) six repeated i.p. injections of the first three concentrations of colchicine, over a period of 18 days, so as to mimic a chronic exposure. Repeated blood samplings were performed by cardiac puncture over a period of about 2 months after the treatment and micronucleus frequencies were determined at multiple times on the same individuals after mutagen exposure. A dose-dependent increase in the micronucleus frequency was observed in irradiated fish and a peak value detected at 21 days. Slight increases of micronucleus frequencies were also observed in both colchicine experiments only for the highest concentrations at the earliest sampling time. Higher concentrations of colchicine clearly showed a lethal effect. According to the present data the micronucleus frequency induced by the highest colchicine dose is comparable to that observed after 0.1Gy of X ray irradiation. PMID- 11423355 TI - Differences in the number of micronucleated erythrocytes among young and adult animals including humans. Spontaneous micronuclei in 43 species. AB - In our previous report we speculated about the possibility that some species had high levels of spontaneous micronucleated erythrocytes (MNE) just in a juvenile stage, this is, that the MNE diminish as the reticuloendothelial system matures. Here we show this effect in species including rat, rabbit, pig, dog, cat, gray squirrel, lion, giraffe, white-tailed deer, opossum and even human. The number of spontaneous MNE that we found in 43 species is shown, and the proportions of polychromatic and normochromatic. This is our third report on spontaneous MNE in different species. We obtained 189 peripheral blood samples of mammals, birds and reptiles. From 12 species we obtained only one sample, and 16 were reported previously, but now the size of the sample has been increased. The species with the highest spontaneous MNE were the Vietnamese potbelly pig (with the highest MNE number), Bengal tiger, capuchin monkey, puma, ferret, owl, hedgehog, squirrel monkey, pig and white-tailed deer. These species could be used as monitors for genotoxic events. PMID- 11423356 TI - Rapid diagnostic techniques for malaria control. AB - The past decade was a milestone in the development of malaria diagnostic technology. Today, a variety of simplified and rapid malaria diagnostic devices, collectively referred to as 'dipsticks', is available. This paper discusses the potential roles of these devices, and obstacles to their use, in supporting malaria control strategies. PMID- 11423357 TI - The paradox of endemic stability. PMID- 11423372 TI - Phospholipid remodeling/generation in Giardia: the role of the Lands cycle. AB - Recent results suggest that Giardia is able to carry out deacylation/reacylation reactions (the Lands cycle) to generate new phospholipids, effectively bypassing the de novo synthesis of the entire phospholipid molecule. The successful operation of this deacylation/reacylation cycle is important for Giardia because this protozoan parasite possesses limited lipid synthesis ability. This article discusses how Giardia might use the Lands cycle to alter phospholipids acquired from the host during its colonization in the human small intestine. PMID- 11423374 TI - Host--parasite relationships of Schistosoma japonicum in mammalian hosts. AB - Control of schistosomiasis caused by Schistosoma japonicum has been severely hindered by the fact that several non-human mammalian species, including domesticated as well as wild animals, serve as zoonotic carriers of this infection. For effective control, it is imperative that the full host spectrum of this infection is understood. Although about 46 species of mammals are known to carry natural infection with S. japonicum, only a few might be of potential threat to human infection. Generally, in an endemic area, transmission of schistosomiasis to human depends largely on the availability and abundance of permissive hosts. Another important factor that needs to be taken into consideration in developing control measures against S. japonicum is potential strain differences. This review collates pertinent host-parasite relationship of S. japonicum in mammals in an endemic area and assesses the epidemiological significance of these findings for human infection. PMID- 11423375 TI - Influence of host nutrition on the development and consequences of nematode parasitism in ruminants. AB - Control of gastrointestinal nematodes of ruminants is based largely on use of anthelmintics combined, where practical, with pasture management. The increasing prevalence of resistance to anthelmintics has led to the search for alternative sustainable control strategies. Here, we consider how nutrition, as a short-term alternative, can influence the host--parasite relationship in ruminants, using gastrointestinal nematode infections of sheep as the model system. Nutrition can affect the ability of the host to cope with the consequences of parasitism and to contain and eventually to overcome parasitism. It can also affect the parasite population through the intake of antiparasitic compounds. PMID- 11423376 TI - Parasite adhesion and immune evasion in placental malaria. AB - Parasite sequestration in the placenta is a key feature of infection by Plasmodium falciparum during pregnancy and is associated with severe adverse outcomes for both mother and baby. Here, James Beeson and colleagues draw together the findings of recent studies on parasite mechanisms that mediate this process. They review evidence for novel parasite variants that appear able to evade pre-existing immunity, for the adhesion of P. falciparum-infected erythrocytes to placental glycosaminoglycans (and the molecular basis of these parasite properties) and for the expression of var genes encoding the variant antigen and adhesive ligand P. falciparum-erythrocyte membrane protein 1 (PfEMP1). PMID- 11423377 TI - An update on antigenic variation in African trypanosomes. AB - African trypanosomes can spend a long time in the blood of their mammalian host, where they are exposed to the immune system and are thought to take advantage of it to modulate their own numbers. Their major immunogenic protein is the variant surface glycoprotein (VSG), the gene for which must be in one of the 20--40 specialized telomeric expression sites in order to be transcribed. Trypanosomes escape antibody-mediated destruction through periodic changes of the expressed VSG gene from a repertoire of approximately 1000. How do trypanosomes exclusively express only one VSG and how do they switch between them? PMID- 11423378 TI - Molecular tools and triatomine systematics: a public health perspective. AB - Triatomines, or kissing bugs, are vectors of Chagas disease to humans. This disease is a substantial public health problem affecting up to 12 million people throughout the Americas, and its control relies mainly on the insecticide treatment of triatomine-infested houses within villages. In this article, Fernando Monteiro, Ananias Escalante and Ben Beard review how molecular markers have been used to clarify triatomine systematics, and give examples of how our understanding of triatomine population structure and accurate vector identification can be used to optimize vector control. PMID- 11423380 TI - Comparative analyses of the pigment-aggregating and -dispersing actions of MCH on fish chromatophores. AB - In melanophores of the peppered catfish and the Nile tilapia, melanin concentrating hormone (MCH) at low doses (<1 microM) induced pigment aggregation, and the aggregated state was maintained in the presence of MCH. However, at higher MCH concentrations (such as 1 and 10 microM), pigment aggregation was immediately followed by some re-dispersion, even in the continued presence of MCH, which led to an apparent decrease in aggregation. This pigment-dispersing activity at higher concentrations of MCH required extracellular Ca(2+) ions. By contrast, medaka melanophores responded to MCH only by pigment aggregation, even at the highest concentration employed (10 microM). Since it is known that medaka melanophores possess specific receptors for alpha-melanophore-stimulating hormone (alpha-MSH), the possibility that interaction between MSH receptors and MCH at high doses in the presence of Ca(2+) might cause pigment dispersion is ruled out. Cyclic MCH analogs, MCH (1-14) and MCH (5-17), failed to induce pigment dispersion, whereas they induced aggregation of melanin granules. These results suggest that another type of MCH receptor that mediates pigment dispersion is present in catfish and tilapia melanophores, and that intact MCH may be the only molecule that can bind to these receptors. Determinations of cAMP content in melanophores, which were isolated from the skin of three fish species and treated with 10 nM or 10 microM MCH, indicate that MCH receptors mediating aggregation may be coupled with Gi protein, whereas MCH receptors that mediate dispersion may be linked to Gs. The response of erythrophores, xanthophores and leucophores to MCH at various concentrations was also examined, and the results suggest that the distribution patterns of the two types of MCH receptors may differ among fish species and among types of chromatophore in the same fish. PMID- 11423381 TI - DL-Buthionine-S,R-sulfoximine affects intestinal alkaline phosphatase activity. AB - The susceptibility of intestinal alkaline phosphatase to DL-buthionine-S,R sulfoximine was investigated in chicks fed a commercial diet. The results show that DL-buthionine-S,R-sulfoximine produced inhibition of intestinal alkaline phosphatase activity. This effect showed dose- and time-dependency and it was caused by either in vivo DL-buthionine-S,R- sulfoximine administration or in vitro DL-buthionine-S,R-sulfoximine incubation with villus tip enterocytes. DL Buthionine-S,R-sulfoximine did not act directly on intestinal alkaline phosphatase but it provoked glutathione depletion which led to changes in the redox state of the enterocyte as shown by the production of free hydroxyl radicals and an incremental increase in the carbonyl content of proteins. The reversibility of the buthionine sulfoximine effect on intestinal alkaline phosphatase was proved by addition of glutathione monoester to the duodenal loop. PMID- 11423382 TI - Accelerated tissue aging and increased oxidative stress in broiler chickens fed allopurinol. AB - Uric acid has been hypothesized as being one of the more important antioxidants in limiting the accumulation of glycosylated endproducts in birds. Study 1 was designed to quantitatively manipulate the plasma concentrations of uric acid using hemin and allopurinol while study 2 determined their effects on skin pentosidine, the shear force value of Pectoralis major muscle, plasma glucose, body weight and chemiluminescence monitored oxidative stress in broiler chickens. Hemin was hypothesized to raise uric acid concentrations thereby lowering oxidative stress whereas allopurinol was hypothesized to lower uric acid concentrations and raise measures of oxidative stress. In study 1 feeding allopurinol (10 mg/kg body weight) to 8-week-old broiler chicks (n=50) for 10 days decreased plasma uric acid by 57%. However, hemin (10 mg/kg body weight) increased uric acid concentrations 20%. In study 2, 12-week-old broiler chicks (n=90) were randomly assigned to either an ad libitum (AL) diet or a diet restricted (DR) group. Each group was further divided into three treatments (control, allopurinol or hemin fed). Unexpectedly, hemin did not significantly effect uric acid concentrations but increased (P<0.05) measures of chemiluminescence dependent oxidative stress in both the DR and AL birds probably due to the ability of iron to generate oxygen radicals. Allopurinol lowered concentrations of uric acid and increased (P<0.05) the oxidative stress in the AL birds at week 22, reduced (P<0.05) body weight in both the AL and DR fed birds at 16 and 22 weeks of age, and markedly increased (P<0.001) shear force values of the pectoralis major muscle. Skin pentosidine levels increased (P<0.05) in AL birds fed allopurinol or hemin fed birds, but not in the diet restricted birds at 22 weeks. The significance of these studies is that concentrations of plasma uric acid can be related to measures of oxidative stress, which can be linked to tissue aging. PMID- 11423383 TI - Effects of Quillaja saponins on growth, metabolism, egg production and muscle cholesterol in individually reared Nile tilapia (Oreochromis niloticus). AB - The effects of supplementation of a Quillaja saponin (QS) mixture in the diets of tilapia have been studied using a respirometer system that allowed feeding and continuous measurement of oxygen consumption of individual fish. Five fish each were given control diet (C group) and control diet supplemented with 150 mg kg( 1) (S150 group) or 300 mg kg(-1) (S300 group) QS. At the end of 14 weeks the weight gain of the S300 group was significantly higher than control (P<0.05) whereas that of the S150 group had an intermediate value. The S150 group had a higher growth rate (P=0.05) after the first 3 weeks of feeding with the experimental diets, compared to the other two groups. At the end of the experiment the S300 group had significantly higher (P<0.05) average values for energy retention, apparent lipid conversion, carcass fat, energy and significantly lower (P<0.05) average values for apparently unutilised energy and carcass ash content compared to the C group. The corresponding values of the S150 group were intermediate between the C and S300 groups. One out of two female fish in the S150 group and both female fish in the S300 group never produced eggs during the entire 14-week experimental period. Contrarily, all three female fish in the control group and one out of the two female fish in the S150 group regularly produced eggs, at a rate of approximately once in every 14 days. The muscle cholesterol level in the S300 group was significantly higher than that of the C group. Possible mechanisms of action of the dietary saponins are discussed. PMID- 11423384 TI - Measurement of cytochrome P4501A induction in dab (Limanda limanda) and other teleosts with species-specific cDNA probes: isolation and characterisation of dab cDNA and its use in expression studies with beta-naphthoflavone-treated fish. AB - Induction of cytochrome P4501A (CYP1A) in fish is an important biomarker in marine monitoring programmes but a number of factors complicate interpretation of data based on catalytic activity. To provide additional analytical tools, we have cloned and sequenced entire (dab) and partial cDNAs (flounder, turbot, sand eel) from several fish species. A detailed analysis comparing the new sequences to those on the database (13 sequences) is presented and identifies an invariant, teleost-specific sequence (195-IVVSVANVICGMCFGRRYDH-214) which might be the basis for production of a species cross-reactive antibody. Northern and slot blots of fish RNA (sand eel, plaice, turbot, flounder and dab) showed extensive cross species hybridisation with each of the cDNAs (sand eel, plaice, turbot, flounder and dab). The exception was turbot RNA, which only gave adequate hybridisation when the turbot probe was used. Attempts to normalise the hybridisation data to GAPDH mRNA were not satisfactory since there were significant species differences in expression of this gene and expression was suppressed (20-40%) by beta naphthoflavone treatment. The CYP1A probes indicated induction levels relative to untreated dab of: plaice (five-fold); turbot (12-fold); flounder (12-fold); and dab (10-fold). The study demonstrates the relative ease with which species specific molecular probes can be generated and used. PMID- 11423385 TI - Effects of cholinergic agents on the vasopressin-mediated water transport in the isolated toad bladder. AB - The aim of this work was to study the effect of some pharmacological cholinergic agents on the events that follow the interaction of arginine vasopressin with toad bladder membrane receptors related to synthesis of 3'5'(c)AMP. The water flow through the membrane was measured gravimetrically in sac preparations of the membrane. In the absence of arginine vasopressin (AVP), carbachol induced a significant increase in the water flow (37%) related to the basal (Ringer's solution). On the other hand, when carbachol and AVP were associated, a significant decrease of AVP hydrosmotic activity occurred (23%). The inhibitory effect of carbachol on the AVP action was almost completely abolished by the cholinergic antagonists atropine, pirenzepine, 4-diphenylacetoxy-N methylpiperidine methiodide (4-DAMP) and the calcium antagonist lanthanum. Similarly, when carbachol and 3'5' cyclic adenosine monophosphate (3'5'(c)AMP) were associated, a decrease of nucleotide hydrosmotic activity was observed (12.80%). This effect was partially restored by the addition of pirenzepine or 4 DAMP in the bath solution. These results suggest a role for muscarinic receptors of sub-type M(1) and M(3), which are involved in the intracellular calcium release. The increase of calcium concentration in the intracellular medium acts as a negative modulator in the hydrosmotic action of antidiuretic hormone. PMID- 11423386 TI - Interaction of estrogen and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) with hepatic fatty acid synthesis and metabolism of male chickens (Gallus domesticus). AB - This study tested the hypothesis that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) antagonizes estrogen-induced hepatic lipid synthesis and metabolism in birds. Twenty immature male chickens (Gallus domesticus) were divided evenly into four groups: (1) vehicle control; (2) estrogen alone (1.0 mg/kg estradiol cypionate injected on three consecutive days); (3) TCDD alone (50 microg/kg injected on the fourth day); and (4) a combination of the estrogen and TCDD treatments. On day 14, liver samples were collected for quantitative fatty acid analysis by capillary gas chromatography. Birds treated with estrogen alone had increased total triacylglyceride concentrations with specific increases in the Delta9 desaturase products 16:1n7, 18:1n7, 18:1n9, and 20:1n9. In addition, estrogen treatment specifically increased 22:6n3 concentrations in both triacylglycerides and phospholipids. However, these increases in Delta9 desaturase products or 22:6n3 did not occur for birds treated with estrogen in combination with TCDD. TCDD and estrogen plus TCDD treatments increased phospholipid concentrations of the diet-derived polyunsaturated fatty acids 18:2n6, 18:3n6, 20:3n6, 18:3n3, and 20:5n3, although only the estrogen plus TCDD group had significantly increased total phospholipids. In cholesterol esters, all three treatments decreased concentrations of total fatty acids, saturated fatty acids, and Delta9 desaturase products compared to the control group. PMID- 11423387 TI - Venom of the crotaline snake Atropoides nummifer (jumping viper) from Guatemala and Honduras: comparative toxicological characterization, isolation of a myotoxic phospholipase A(2) homologue and neutralization by two antivenoms. AB - A comparative study was performed on the venoms of the crotaline snake Atropoides nummifer from Guatemala and Honduras. SDS-polyacrylamide gel electrophoresis, under reducing conditions, revealed a highly similar pattern of these venoms, and between them and the venom of the same species from Costa Rica. Similar patterns were also observed in ion-exchange chromatography on CM-Shephadex C-25, in which a highly basic myotoxic fraction was present. This fraction was devoid of phospholipase A(2) activity and strongly reacted, by enzyme-immunoassay, with an antiserum against Bothrops asper myotoxin II, a Lys-49 phospholipase A(2) homologue. A basic myotoxin of 16 kDa was isolated to homogeneity from the venom of A. nummifer from Honduras, showing amino acid composition and N-terminal sequence similar to those of Lys-49 phospholipase A(2) variants previously isolated from other crotaline snake venoms. Guatemalan and Honduran A. nummifer venoms have a qualitatively similar toxicological profile, characterized by: lethal; hemorrhagic; myotoxic; edema-forming; coagulant; and defibrinating activities, although there were significant quantitative variations in some of these activities between the two venoms. Neutralization of toxic activities by two commercially-available antivenoms in the region was studied. Polyvalent antivenom produced by Instituto Clodomiro Picado was effective in the neutralization of: lethal; hemorrhagic; myotoxic; coagulant; defibrinating; and phospholipase A(2) activities, but ineffective against edema-forming activity. On the other hand, MYN polyvalent antivenom neutralized: hemorrhagic; myotoxic; coagulant; defibrinating; and phospholipase A(2) activities, albeit with a lower potency than Instituto Clodomiro Picado antivenom. MYN antivenom failed to neutralize lethal and edema-forming activities of A. nummifer venoms. PMID- 11423388 TI - Effect of vitamin E and selenium on resistance to oxidative stress in chicken superficial pectoralis muscle. AB - One-day-old chicks were reared using diets that differed in their vitamin E and/or selenium content. In chicks depleted of both selenium and vitamin E, signs of exudative diathesis on the superficial pectoralis muscle were observed. The purpose of this research was to determine the defective points of the antioxidant defense system, which made this tissue highly susceptible to nutritionally induced oxidative stress. Vitamin E, and selenium in lower magnitude, were the factors that strikingly affected the course of mitochondrial lipid peroxidation. Animals fed diets deficient in vitamin E and selenium displayed the lowest reduced glutathione level and glutathione peroxidase activity. The decreased levels of reduced glutathione were not due to a defective activity of glutathione reductase, which was increased in both mitochondria and cytosol. The absence of vitamin E was linked to lowering of mitochondrial thiol levels. The Glutathione peroxidase/Cu,Zn-superoxide dismutase ratio was 2.8 in animals fed selenium and vitamin E, and decreased to 0.13 in animals deficient in both nutrients. This change was indicative of oxidant-induced damage mediated by hydrogen peroxide. Catalase activity increased in an attempt to counteract the decrease in glutathione peroxidase activity. The results obtained showed that alpha tocopherol and Se deficiencies caused multiple alterations in the antioxidant system and adversely affected the redox state of chicken superficial pectoralis muscle. PMID- 11423389 TI - No substantial difference in genotype frequencies of interleukin and myeloperoxidase polymorphisms between malignant lymphoma patients and non-cancer controls. AB - BACKGROUND AND OBJECTIVES: The functional polymorphisms regulating immunologic responses may influence the proliferation or suppression of malignant lymphoma. We examined the association between malignant lymphoma risk and the polymorphisms of the IL-1 gene family [IL-1B -31 C/T, IL-1A -889 C/T, and IL-1RN 86-bp variable number of terminal repeat (VNTR)] and myeloperoxidase (MPO -463 G/A). DESIGN AND METHODS: The hospital-based case-control study was conducted in Japan. Genotypes were examined in a total of 372 lymphoma cases and 241 non-cancer control subjects. The relative risks were estimated by unconditional logistic regression analysis. RESULTS: The overall allele distribution of these polymorphisms did not differ substantially between patients and controls; the odds ratios were 0.73 (95% confidence interval, 0.48-1.11) for the T allele carriers of IL-1B relative to the non-carriers, 1.01 (0.56-1.82) for the 2-repeat allele (allele 2) carriers of IL-1RN, 0.96 (0.62-1.48) for the T allele carriers of IL-1A, and 1.04 (0.70 1.57) for the A allele carriers of MPO. Subgroup analyses according to histology [diffuse large B-cell lymphoma (DLBL), follicular lymphoma, low-grade lymphoma of mucosa associated lymphoid tissue, and others] failed to illustrate differences except for DLBL which showed a possible association with IL-1A and IL-1B polymorphisms. INTERPRETATION AND CONCLUSIONS: Our data show a limited association between these polymorphisms and malignant lymphoma risk in total. The possible association of the IL-1A and IL-1B polymorphisms with DLB-needs further clarification. PMID- 11423390 TI - The functional form of angular forces around transition metal ions in biomolecules. AB - A method for generating angular forces around sigma-bonded transition metal ions is generalized to treat pi-bonded configurations. The theoretical approach is based on an analysis of ligand-field and small-cluster Hamiltonians based on the moments of the electron state distribution. The functional forms that are obtained involve a modification of the usual expression of the binding energy as a sum of ligand-ligand interactions, which, however, requires very little increase in CPU time. The angular interactions have simple forms involving sin and cos functions, whose relative weights depend on whether the ligands are sigma or pi-bonded. They describe the ligand-field stabilization energy to an accuracy of about 10%, and the interaction energy of covalently bonded systems to an accuracy of better than 4%. The resulting functional forms for the force field are used to model the structure of small clusters, including fragments of the copper blue protein structure. Large deviations from the typical square copper coordination are found when pi-bonded ligands are present. PMID- 11423391 TI - Simulation of motor-driven cochlear outer hair cell electromotility. AB - We propose a three-dimensional (3D) model to simulate outer hair cell electromotility. In our model, the major components of the composite cell wall are explicitly represented. We simulate the activity of the particles/motor complexes in the plasma membrane by generating active strains inside them and compute the overall response of the cell. We also consider the constrained wall and compute the generated active force. We estimate the parameters of our model by matching the predicted longitudinal and circumferential electromotile strains with those observed in the microchamber experiment. In addition, we match the earlier estimated values of the active force and cell wall stiffness. The computed electromotile strains in the plasma membrane and other components of the wall are in agreement with experimental observations in trypsinized cells and in nonmotile cells transfected with Prestin. We discover several features of the 3D mechanism of outer hair cell electromotilty. Because of the constraints under which the motors operate, the motor-related strains have to be 2-3 times larger than the observable strains. The motor density has a strong effect on the electromotile strain. Such effect on the active force is significantly lower because of the interplay between the active and passive properties of the cell wall. PMID- 11423392 TI - Diffusion of microspheres in shear flow near a wall: use to measure binding rates between attached molecules. AB - The rate and distance-dependence of association between surface-attached molecules may be determined by monitoring the motion of receptor-bearing spheres along ligand-coated surfaces in a flow chamber (Pierres et al., Proc. Natl. Acad. Sci. U.S.A. 95:9256-9261, 1998). Particle arrests reveal bond formation, and the particle-to-surface distance may be estimated from the ratio between the velocity and the wall shear rate. However, several problems are raised. First, data interpretation requires extensive computer simulations. Second, the relevance of standard results from fluid mechanics to micrometer-size particles separated from surfaces by nanometer distances is not fully demonstrated. Third, the wall shear rate must be known with high accuracy. Here we present a simple derivation of an algorithm permitting one to simulate the motion of spheres near a plane in shear flow. We check that theoretical predictions are consistent with the experimental dependence of motion on medium viscosity or particle size, and the requirement for equilibrium particle height distribution to follow Boltzman's law. The determination of the statistical relationship between particle velocity and acceleration allows one to derive the wall shear rate with 1-s(-1) accuracy and the Hamaker constant of interaction between the particle and the wall with a sensitivity better than 10(-21) J. It is demonstrated that the correlation between particle height and mean velocity during a time interval Deltat is maximal when Deltat is about 0.1-0.2 s for a particle of 1.4-microm radius. When the particle-to-surface distance ranges between 10 and 40 nm, the particle height distribution may be obtained with a standard deviation ranging between 8 and 25 nm, provided the average velocity during a 160-ms period of time is determined with 10% accuracy. It is concluded that the flow chamber allows one to detect the formation of individual bonds with a minimal lifetime of 40 ms in presence of a disruptive force of approximately 5 pN and to assess the distance dependence within the tens of nanometer range. PMID- 11423393 TI - Direct measurement of the area expansion and shear moduli of the human red blood cell membrane skeleton. AB - The area expansion and the shear moduli of the free spectrin skeleton, freshly extracted from the membrane of a human red blood cell (RBC), are measured by using optical tweezers micromanipulation. An RBC is trapped by three silica beads bound to its membrane. After extraction, the skeleton is deformed by applying calibrated forces to the beads. The area expansion modulus K(C) and shear modulus mu(C) of the two-dimensional spectrin network are inferred from the deformations measured as functions of the applied stress. In low hypotonic buffer (25 mOsm/kg), one finds K(C) = 4.8 +/- 2.7 microN/m, mu(C) = 2.4 +/- 0.7 microN/m, and K(C)/mu(C) = 1.9 +/- 1.0. In isotonic buffer, one measures higher values for K(C), mu(C), and K(C)/mu(C), partly because the skeleton collapses in a high ionic-strength environment. Some data concerning the time evolution of the mechanical properties of the skeleton after extraction and the influence of ATP are also reported. In the Discussion, it is shown that the measured values are consistent with estimates deduced from experiments carried out on the intact membrane and agree with theoretical and numerical predictions concerning two dimensional networks of entropic springs. PMID- 11423394 TI - Membrane-initiated Ca(2+) signals are reshaped during propagation to subcellular regions. AB - An important aspect of Ca(2+) signaling is the ability of cells to generate intracellular Ca(2+) waves. In this study we have analyzed the cellular and subcellular kinetics of Ca(2+) waves in a neuroendocrine transducer cell, the melanotrope of Xenopus laevis, using the ratiometric Ca(2+) probe indo-1 and video-rate UV confocal laser-scanning microscopy. The purpose of the present study was to investigate how local Ca(2+) changes contribute to a global Ca(2+) signal; subsequently we quantified how a Ca(2+) wave is kinetically reshaped as it is propagated through the cell. The combined kinetics of all subcellular Ca(2+) signals determined the shape of the total cellular Ca(2+) signal, but each subcellular contribution to the cellular signal was not constant in time. Near the plasma membrane, [Ca(2+)](i) increased and decreased rapidly, processes that can be described by a linear and exponential function, respectively. In more central parts of the cell slower kinetics were observed that were best described by a Hill equation. This reshaping of the Ca(2+) wave was modeled with an equation derived from a low-pass RC filter. We propose that the differences in spatial kinetics of the Ca(2+) signal serves as a mechanism by which the same cellular Ca(2+) signal carries different regulatory information to different subcellular regions of the cell, thus evoking differential cellular responses. PMID- 11423395 TI - Microtubule treadmilling in vitro investigated by fluorescence speckle and confocal microscopy. AB - Whether polarized treadmilling is an intrinsic property of microtubules assembled from pure tubulin has been controversial. We have tested this possibility by imaging the polymerization dynamics of individual microtubules in samples assembled to steady-state in vitro from porcine brain tubulin, using a 2% glycerol buffer to reduce dynamic instability. Fluorescence speckled microtubules were bound to the cover-glass surface by kinesin motors, and the assembly dynamics of plus and minus ends were recorded with a spinning-disk confocal fluorescence microscopy system. At steady-state assembly, 19% of the observed microtubules (n = 89) achieved treadmilling in a plus-to-minus direction, 34% in a minus-to-plus direction, 37% grew at both ends, and 10% just shortened. For the population of measured microtubules, the distribution of lengths remained unchanged while a 20% loss of original and 27% gain of new polymer occurred over the 20-min period of observation. The lack of polarity in the observed treadmilling indicates that stochastic differences in dynamic instability between plus and minus ends are responsible for polymer turnover at steady-state assembly, not unidirectional treadmilling. A Monte Carlo simulation of plus and minus end dynamics using measured dynamic instability parameters reproduces our experimental results and the amount of steady-state polymer turnover reported by previous biochemical assays. PMID- 11423396 TI - Interaction of SNX482 with domains III and IV inhibits activation gating of alpha(1E) (Ca(V)2.3) calcium channels. AB - We have investigated the action of SNX482, a toxin isolated from the venom of the tarantula Hysterocrates gigas, on voltage-dependent calcium channels expressed in tsa-201 cells. Upon application of 200 nM SNX482, R-type alpha(1E) calcium channels underwent rapid and complete inhibition, which was only poorly reversible upon washout. However, upon application of strong membrane depolarizations, rapid and complete recovery from inhibition was obtained. Tail current analysis revealed that SNX482 mediated an approximately 70 mV depolarizing shift in half-activation potential, suggesting that the toxin inhibits alpha(1E) calcium channels by preventing their activation. Experiments involving chimeric channels combining structural features of alpha(1E) and alpha(1C) subunits indicated that the presence of the domain III and IV of alpha(1E) is a prerequisite for a strong gating inhibition. In contrast, L-type alpha(1C) channels underwent incomplete inhibition at saturating concentrations of SNX482 that was paralleled by a small shift in half-activation potential and which could be rapidly reversed, suggesting a less pronounced effect of the toxin on L-type calcium channel gating. We conclude that SNX482 does not exhibit unequivocal specificity for R-type channels, but highly effectively antagonizes their activation. PMID- 11423397 TI - Ca(2+) channel inactivation heterogeneity reveals physiological unbinding of auxiliary beta subunits. AB - Voltage gated Ca(2+) channel (VGCC) auxiliary beta subunits increase membrane expression of the main pore-forming alpha(1) subunits and finely tune channel activation and inactivation properties. In expression studies, co-expression of beta subunits also reduced neuronal Ca(2+) channel regulation by heterotrimeric G protein. Biochemical studies suggest that VGCC beta subunits and G protein betagamma can compete for overlapping interaction sites on VGCC alpha(1) subunits, suggesting a dynamic association of these subunits with alpha(1). In this work we have analyzed the stability of the alpha(1)/beta association under physiological conditions. Regulation of the alpha(1A) Ca(2+) channel inactivation properties by beta(1b) and beta(2a) subunits had two major effects: a shift in voltage-dependent inactivation (E(in)), and an increase of the non-inactivating current (R(in)). Unexpectedly, large variations in magnitude of the effects were recorded on E(in), when beta(1b) was expressed, and R(in), when beta(2a) was expressed. These variations were not proportional to the current amplitude, and occurred at similar levels of beta subunit expression. beta(2a)-induced variations of R(in) were, however, inversely proportional to the magnitude of G protein block. These data underline the two different mechanisms used by beta(1b) and beta(2a) to regulate channel inactivation, and suggest that the VGCC beta subunit can unbind the alpha1 subunit in physiological situations. PMID- 11423398 TI - Modification of voltage-dependent gating of potassium channels by free form of tryptophan side chain. AB - Indole constitutes a major component of the side chain of the amino acid tryptophan. Application of indole slows activation of voltage-dependent potassium channels and reduces steady-state conductance in a voltage- and concentration dependent manner. The steep concentration dependence indicates that multiple indole molecules may interact with the channel. Indole does not noticeably change the unitary conductance or the mean open duration, however, it accelerates off gating currents without altering on-gating currents. These properties of the modification of channel gating induced by indole are consistent with a model in which indole binds independently to every subunit of the channel complex to prevent the final concerted transition to the open state. We suggest that exogenously applied indole and side-chains of the tryptophan residues of the channel protein involved in activation may compete for the same effector position and that indole might be useful as a probe to study functional roles of tryptophan residues. PMID- 11423399 TI - High-affinity Zn block in recombinant N-methyl-D-aspartate receptors with cysteine substitutions at the Q/R/N site. AB - In ionotropic glutamate receptors, many channel properties (e.g., selectivity, ion permeation, and ion block) depend on the residue (glutamine, arginine, or asparagine) located at the tip of the pore loop (the Q/R/N site). We substituted a cysteine for the asparagine present at that position in both NR1 and NR2 N methyl-D-aspartate (NMDA) receptor subunits. Under control conditions, receptors containing mutated NR1 and NR2 subunits show much smaller glutamate responses than wild-type receptors. However, this difference disappears upon addition of heavy metal chelators in the extracellular bath. The presence of cysteines at the Q/R/N site in both subunits of NR1/NR2C receptors results in a 220,000-fold increase in sensitivity of the inhibition by extracellular Zn. In contrast with the high-affinity Zn inhibition of wild-type NR1/NR2A receptors, the high affinity Zn inhibition of mutated NR1/NR2C receptors shows a voltage dependence, which resembles very much that of the block by extracellular Mg. This indicates that the Zn inhibition of the mutated receptors results from a channel block involving Zn binding to the thiol groups introduced into the selectivity filter. Taking advantage of the slow kinetics of the Zn block, we show that both blocking and unblocking reactions require prior opening of the channel. PMID- 11423400 TI - Single-channel recordings of recombinant inositol trisphosphate receptors in mammalian nuclear envelope. AB - Inositol 1,4,5-trisphosphate (InsP(3)) receptors (InsP(3)Rs) are intracellular Ca(2+) channels gated by the second messenger InsP(3). Here we describe a novel approach for recording single-channel currents through recombinant InsP(3)Rs in mammalian cells that applies patch-clamp electrophysiology to nuclei isolated from COS-7 cells transiently transfected with the neuronal (SII(+)) and peripheral (SII(-)) alternatively-spliced variants of the rat type 1 InsP(3)R. Single channels that were activated by InsP(3) and inhibited by heparin were observed in 45% of patches from nuclei prepared from transfected cells overexpressing recombinant InsP(3)Rs. In contrast, nuclei from cells transfected with the vector alone had InsP(3)-dependent channel activity in only 1.5% of patches. With K(+) (140 mM) as the permeant ion, recombinant SII(+) and SII(-) channels had slope conductances of 370 pS and 390 pS, respectively. The recombinant channels were 4-fold more selective for Ca(2+) over K(+), and their open probabilities were biphasically regulated by cytoplasmic [Ca(2+)]. This approach provides a powerful new methodology to study the permeation and gating properties of recombinant mammalian InsP(3)Rs in a native mammalian membrane environment at the single-channel level. PMID- 11423401 TI - Modulation of Kv1.5 potassium channel gating by extracellular zinc. AB - Zinc ions are known to induce a variable depolarizing shift of the ionic current half-activation potential and substantially slow the activation kinetics of most K(+) channels. In Kv1.5, Zn(2+) also reduces ionic current, and this is relieved by increasing the external K(+) or Cs(+) concentration. Here we have investigated the actions of Zn(2+) on the gating currents of Kv1.5 channels expressed in HEK cells. Zn(2+) shifted the midpoint of the charge-voltage (Q-V) curve substantially more (approximately 2 times) than it shifted the V(1/2) of the g-V curve, and this amounted to +60 mV at 1 mM Zn(2+). Both Q1 and Q2 activation charge components were similarly affected by Zn(2+), which indicated free access of Zn(2+) to channel closed states. The maximal charge movement was also reduced by 1 mM Zn(2+) by approximately 15%, from 1.6 +/- 0.5 to 1.4 +/- 0.47 pC (n = 4). Addition of external K(+) or Cs(+), which relieved the Zn(2+)-induced ionic current reduction, decreased the extent of the Zn(2+)-induced Q-V shift. In 135 mM extracellular Cs(+), 200 microM Zn(2+) reduced ionic current by only 8 +/- 1%, compared with 71% reduction in 0 mM extracellular Cs(+), and caused a comparable shift in both the g-V and Q-V relations (17.9 +/- 0.6 mV vs. 20.8 +/- 2.1 mV, n = 6). Our results confirm the presence of two independent binding sites involved in the Zn(2+) actions. Whereas binding to one site accounts for reduction of current and binding to the other site accounts for the gating shift in ionic current recordings, both sites contribute to the Zn(2+)-induced Q-V shift. PMID- 11423402 TI - Gating properties of gap junction channels assembled from connexin43 and connexin43 fused with green fluorescent protein. AB - We used cell lines expressing wild-type connexin43 (Cx43) and Cx43 fused with enhanced green fluorescent protein (Cx43-EGFP) to examine mechanisms of gap junction channel gating. Previously it was suggested that each hemichannel in a cell-cell channel possesses two gates, a fast gate that closes channels to a nonzero conductance or residual state via fast (< approximately 2 ms) transitions and a slow gate that fully closes channels via slow transitions (> approximately 10 ms). Here we demonstrate that transjunctional voltage (V(j)) regulates both gates and that they are operating in series and in a contingent manner in which the state of one gate affects gating of the other. Cx43-EGFP channels lack fast V(j) gating to a residual state but show slow V(j) gating. Both Cx43 and Cx43 EGFP channels exhibit slow gating by chemical uncouplers such as CO(2) and alkanols. Chemical uncouplers do not induce obvious changes in Cx43-EGFP junctional plaques, indicating that uncoupling is not caused by dispersion or internalization of junctional plaques. Similarity of gating transitions during chemical gating and slow V(j) gating suggests that both gating mechanisms share common structural elements. Cx43/Cx43-EGFP heterotypic channels showed asymmetrical V(j) gating with fast transitions between open and residual states only when the Cx43 side was relatively negative. This result indicates that the fast V(j) gate of Cx43 hemichannels closes for relative negativity at its cytoplasmic end. PMID- 11423403 TI - Interaction of lung surfactant proteins with anionic phospholipids. AB - Langmuir isotherms, fluorescence microscopy, and atomic force microscopy were used to study lung surfactant specific proteins SP-B and SP-C in monolayers of dipalmitoylphosphatidylglycerol (DPPG) and palmitoyloleoylphosphatidylglycerol (POPG), which are representative of the anionic lipids in native and replacement lung surfactants. Both SP-B and SP-C eliminate squeeze-out of POPG from mixed DPPG/POPG monolayers by inducing a two- to three-dimensional transformation of the fluid-phase fraction of the monolayer. SP-B induces a reversible folding transition at monolayer collapse, allowing all components of surfactant to remain at the interface during respreading. The folds remain attached to the monolayer, are identical in composition and morphology to the unfolded monolayer, and are reincorporated reversibly into the monolayer upon expansion. In the absence of SP B or SP-C, the unsaturated lipids are irreversibly lost at high surface pressures. These morphological transitions are identical to those in other lipid mixtures and hence appear to be independent of the detailed lipid composition of the monolayer. Instead they depend on the more general phenomena of coexistence between a liquid-expanded and liquid-condensed phase. These three-dimensional monolayer transitions reconcile how lung surfactant can achieve both low surface tensions upon compression and rapid respreading upon expansion and may have important implications toward the optimal design of replacement surfactants. The overlap of function between SP-B and SP-C helps explain why replacement surfactants lacking in one or the other proteins often have beneficial effects. PMID- 11423404 TI - Effects of phospholipid unsaturation on the membrane/water interface: a molecular simulation study. AB - Molecular dynamics (MD) simulations of fully hydrated bilayers in the liquid crystalline state made of 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) or 1 palmitoyl-2-elaidoyl-phosphatidylcholine (PEPC) were carried out to investigate the effect of the incorporation of a double bond in the phosphatidylcholine (PC) beta-chain (cis or trans) on the membrane/water interface. The bilayers reached thermal equilibrium after 3 and 1 ns of MD simulations, respectively, and productive runs were carried out for 3 ns for each bilayer. As reference systems, the 1,2-dimyristoyl-phosphatidylcholine (DMPC) bilayer (M. Pasenkiewicz-Gierula, Y. Takaoka, H. Miyagawa, K. Kitamura, and A. Kusumi, 1999, Biophys. J. 76:1228 1240) and DMPC-cholesterol (Chol) bilayer containing 22 mol % Chol (M. Pasenkiewicz-Gierula, T. Rog, K. Kitamura, A. and Kusumi, 2000, Biophys. J. 78:1376-1389) were used. The study shows that at the interface of POPC, PEPC, and DMPC-Chol bilayers, average numbers of PC-water and PC-PC interactions are similar and, respectively, greater and smaller than in the DMPC bilayer. The average area/PC in mono-unsaturated bilayers is approximately 4 A(2) larger than in the DMPC bilayer; nevertheless, a strong correlation was found between a single molecular area (SMA) of a PC and the number of interactions this PC makes; i.e., PCs (either saturated or unsaturated) with the same SMA form similar numbers of intermolecular links. The numbers and corresponding SMAs are distributed about averages pertinent to each bilayer. No significant difference between cis and trans bonds was found. PMID- 11423405 TI - Membrane stress and permeabilization induced by asymmetric incorporation of compounds. AB - The area balance or imbalance between the inner and outer monolayer of biological membranes is a key parameter for driving shape changes (including exo and endocytosis) and controlling the bilayer curvature stress. The asymmetric incorporation of a drug or biological agent interferes with these processes, and the subsequent stress may lead to a membrane permeation or permeabilization. A main goal of this study is to introduce new methods to characterize such phenomena using isothermal titration calorimetry. POPC unilamellar vesicles and a series of alkyl maltosides are used as model systems; the unilamellarity was checked by NMR with the shift reagent Pr(3+). The free energy, enthalpy, and entropy associated with the asymmetry stress are estimated by comparing partitioning data of uptake versus release assays. The asymmetry stress is of enthalpic nature and somewhat reduced by entropic effects. Stimulated membrane permeation occurs at a mean maltoside-to-lipid ratio of approximately 0.2, which corresponds to an apparent area asymmetry of approximately 30% and a limiting free energy of the order of 2 kJ/mol of maltoside. Membrane solubilization to coexisting micelles proceeds at mole ratios of approximately 0.73, 0.81, and 0.88 (C(12)-, C(13)-, and C(14)-maltoside, respectively). Experiments with vesicles pre-loaded with surfactant in both monolayers provide evidence that the translocation threshold is controlled by the asymmetrically incorporated surfactant, whereas the onset of solubilization depends on the total surfactant content in the membrane. Free copies of the uptake and release fitting script including instructions are available upon request to heerklotz@gmx.net. PMID- 11423406 TI - Vesicle trafficking and cell surface membrane patchiness. AB - Membrane proteins and lipids often appear to be distributed in patches on the cell surface. These patches are often assumed to be membrane domains, arising from specific molecular associations. However, a computer simulation (Gheber and Edidin, 1999) shows that membrane patchiness may result from a combination of vesicle trafficking and dynamic barriers to lateral mobility. The simulation predicts that the steady-state patches of proteins and lipids seen on the cell surface will decay if vesicle trafficking is inhibited. To test this prediction, we compared the apparent sizes and intensities of patches of class I HLA molecules, integral membrane proteins, before and after inhibiting endocytic vesicle traffic from the cell surface, either by incubation in hypertonic medium or by expression of a dominant-negative mutant dynamin. As predicted by the simulation, the apparent sizes of HLA patches increased, whereas their intensities decreased after endocytosis and vesicle trafficking were inhibited. PMID- 11423407 TI - Structural properties of a highly polyunsaturated lipid bilayer from molecular dynamics simulations. AB - The structure of a fully hydrated mixed (saturated/polyunsaturated) chain lipid bilayer in the biologically relevant liquid crystalline phase has been examined by performing a molecular dynamics study. The model membrane, a 1-stearoyl-2 docosahexaenoyl-sn-glycero-3-phosphocholine (SDPC, 18:0/22:6 PC) lipid bilayer, was investigated at constant (room) temperature and (ambient) pressure, and the results obtained in the nanosecond time scale reproduced quite well the available experimental data. Polyunsaturated fatty acids are found in high concentrations in neuronal and retinal tissues and are essential for the development of human brain function. The docosahexaenoic fatty acid, in particular, is fundamental for the proper function of the visual receptor rhodopsin. The lipid bilayer order has been investigated through the orientational order parameters. The water-lipid interface has been explored thoroughly in terms of its dimensions and the organization of the different components. Several types of interactions occurring in the system have been analyzed, specifically, the water-hydrocarbon chain, lipid-lipid and lipid-water interactions. The distribution of dihedral angles along the chains and the molecular conformations of the polyunsaturated chain of the lipids have also been studied. Special attention has been focused on the microscopic (molecular) origin of the effects of polyunsaturations on the different physical properties of membranes. PMID- 11423408 TI - Fast lipid disorientation at the onset of membrane fusion revealed by molecular dynamics simulations. AB - Membrane fusion is a key event in vesicular trafficking in every cell, and many fusion-related proteins have been identified. However, how the actual fusion event occurs has not been elucidated. By using molecular dynamics simulations we found that when even a small region of two membranes is closely apposed such that only a limited number of water molecules remain in the apposed area (e.g., by a fusogenic protein and thermal membrane fluctuations), dramatic lipid disorientation results within 100 ps-2 ns, which might initiate membrane fusion. Up to 12% of phospholipid molecules in the apposing layers had their alkyl chains outside the hydrophobic region, lying almost parallel to the membrane surface or protruding out of the bilayer by 2 ns after two membranes were closely apposed. PMID- 11423409 TI - Crystallization screens: compatibility with the lipidic cubic phase for in meso crystallization of membrane proteins. AB - The in meso method for growing crystals of membrane proteins uses a spontaneously forming lipidic cubic mesophase. The detergent-solubilized protein is dispersed with lipid, typically monoolein, and in so doing the cubic phase self-assembles. A precipitant is added to trigger crystal nucleation and growth. The commercial screen solution series are convenient for use in crystallization trials. The aim of this study was to determine which of the Hampton Screen and Screen 2 series of solutions are compatible with the in meso method. These screens contain components any of which could destroy the cubic phase. X-ray diffraction was used for phase identification and for microstructure characterization. The study was done at 4 degrees C and at 20 degrees C. Two types of sample preparations were examined. One used an excess of half-strength screen solution (Prep. 1). The other used a limiting quantity of undiluted screen solution (Prep. 2). At 20 degrees C, over 90% of the screen solutions produced the cubic phase with Prep. 1. This figure dropped to 50% with Prep. 2. In contrast, 50 to 60% of the screens were cubic phase compatible at 4 degrees C under Prep. 1 conditions. The figure fell to 25% with Prep. 2. The mode of action of the diverse screen components are explained on the basis of the phase properties of the monoolein/water system. PMID- 11423410 TI - The influence of lysolipids on the spontaneous curvature and bending elasticity of phospholipid membranes. AB - The effects of lysolipids on phospholipid layer curvature and bending elasticity were examined using x-ray diffraction and the osmotic stress method. Lysolipids with two different head groups, phosphatidylcholine (PC) and phosphatidylethanolamine (PE), and differing hydrocarbon chains were mixed with the hexagonal-forming lipid, dioleoylphosphatidylethanolamine (DOPE). With up to 30 mole% lysolipid in DOPE, the mixture maintains the inverted hexagonal (H(II)) phase in excess water, where increasing levels of lysolipid result in a systematic increase in the H(II) lattice dimension. Analysis of the structural changes imposed by lysolipids show that, opposite to DOPE itself, which has an spontaneous radius of curvature (R(0)) of -30 A, PC lysolipids add high positive curvature, with R(0) = +38 to +60 A, depending on chain length. LysoPEs, in contrast, add very small curvatures. When both polar group and hydrocarbon chains of the added lysolipid mismatch those of DOPE, the structural effects are qualitatively different from otherwise. Such mismatched lysolipids "reshape" the effective combination molecule into a longer and more cylindrical configuration compared to those lysolipids with either matching polar group or hydrocarbon chain. PMID- 11423411 TI - Europium III binding and the reorientation of magnetically aligned bicelles: insights from deuterium NMR spectroscopy. AB - Solid-state deuterium ((2)H) NMR spectroscopy was used to study the reorientation of magnetically ordered bicelles in the presence of the paramagnetic lanthanide Eu(3+). Bicelles were composed of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) plus 1,2-dihexanoyl-sn-glycero-3-phosphocholine plus either the anionic lipid 1,2-dimyristoyl-sn-3-phosphoglycerol, or the cationic lipid 1,2-dimyristoyl 3-trimethyl ammonium propane. Alignment of the bicelles in the magnetic field produced (2)H NMR spectra consisting of a pair of quadrupole doublets, one from the alpha-deuterons and one from the beta-deuterons of DMPC-alpha,beta-d(4). Eu(3+) addition induced the appearance of a second set of quadrupole doublets, having approximately twice the quadrupolar splittings of the originals, and growing progressively in intensity with increasing Eu(3+), at the expense of the intensity of the originals. The new resonances were attributed to bicelles having a parallel alignment with respect to the magnetic field, as opposed to the perpendicular alignment preferred in the absence of Eu(3+). Therefore, the equilibrium degree and kinetics of reorientation could be evaluated from the (2)H NMR spectra. For more cationic initial surface charges, higher amounts of added Eu(3+) were required to induce a given degree of reorientation. However, the equilibrium degree of bicellar reorientation was found to depend solely on the amount of bound Eu(3+), regardless of the bicelle composition. The kinetics of reorientation were a function of lipid concentration. At high lipid concentration, a single fast rate of reorientation (minutes) described the approach to the equilibrium degree of orientation. At lower lipid concentrations, two rates processes were discernible: one fast (minutes) and one slow (hours). The data indicate, therefore, that bicelle reorientation is a phase transition made critical by bicelle-bicelle interactions. PMID- 11423412 TI - Reconstituted syntaxin1a/SNAP25 interacts with negatively charged lipids as measured by lateral diffusion in planar supported bilayers. AB - According to the soluble N-ethylmaleimide-sensitive factor (NSF)-attachment protein (SNAP) receptor hypothesis (SNARE hypothesis), interactions between target SNAREs and vesicle SNAREs (t- and v-SNAREs) are required for membrane fusion in intracellular vesicle transport and exocytosis. The precise role of the SNAREs in tethering, docking, and fusion is still disputed. Biophysical measurements of SNARE interactions in planar supported membranes could potentially resolve some of the key questions regarding the mechanism of SNARE mediated membrane fusion. As a first step toward this goal, recombinant syntaxin1A/SNAP25 (t-SNARE) was reconstituted into polymer-supported planar lipid bilayers. Reconstituted t-SNAREs in supported bilayers bound soluble green fluorescent protein/vesicle-associated membrane protein (v-SNARE), and the SNARE complexes could be specifically dissociated by NSF/alpha-SNAP in the presence of ATP. The physiological activities of SNARE complex formation were thus well reproduced in this reconstituted planar model membrane system. A large fraction (~75%) of the reconstituted t-SNARE was laterally mobile with a lateral diffusion coefficient of 7.5 x 10(-9) cm(2)/s in a phosphatidylcholine lipid background. Negatively charged lipids reduced the mobile fraction of the t-SNARE and the lipids themselves. Phosphatidylinositol-4,5-bisphosphate was more effective than phosphatidylserine in reducing the lateral mobility of the complexes. A model of how acidic lipid-SNARE interactions might alter lipid fluidity is discussed. PMID- 11423413 TI - Lipid-mediated interactions between intrinsic membrane proteins: dependence on protein size and lipid composition. AB - The present study is an application of an approach recently developed by the authors for describing the structure of the hydrocarbon chains of lipid-bilayer membranes (LBMs) around embedded protein inclusions ( Biophys. J. 79:2867-2879). The approach is based on statistical mechanical integral equation theories developed for the study of dense liquids. First, the configurations extracted from molecular dynamics simulations of pure LBMs are used to extract the lateral density-density response function. Different pure LBMs composed of different lipid molecules were considered: dioleoyl phosphatidylcholine (DOPC), palmitoyl oleoyl phosphatidylcholine (POPC), dipalmitoyl phosphatidylcholine (DPPC), and dimyristoyl phosphatidylcholine (DMPC). The results for the lateral density density response function was then used as input in the integral equation theory. Numerical calculations were performed for protein inclusions of three different sizes. For the sake of simplicity, protein inclusions are represented as hard smooth cylinders excluding the lipid hydrocarbon core from a small cylinder of 2.5 A radius, corresponding roughly to one aliphatic chain, a medium cylinder of 5 A radius, corresponding to one alpha-helix, and a larger cylinder of 9 A radius, representing a small protein such as the gramicidin channel. The lipid mediated interaction between protein inclusions was calculated using a closed form expression for the configuration-dependent free energy. This interaction was found to be repulsive at intermediate range and attractive at short range for two small cylinders in POPC, DPPC, and DMPC bilayers, whereas it oscillates between attractive and repulsive values in DOPC bilayers. For medium size cylinders, it is again repulsive at intermediate range and attractive at short range, but for every model LBM considered here. In the case of a large cylinder, the lipid mediated interaction was shown to be repulsive for both short and long ranges for the DOPC, POPC, and DPPC bilayers, whereas it is again repulsive and attractive for DMPC bilayers. The results indicate that the packing of the hydrocarbon chains around protein inclusions in LBMs gives rise to a generic (i.e., nonspecific) lipid-mediated interaction which favors the association of two alpha helices and depends on the lipid composition of the membrane. PMID- 11423414 TI - Interaction of synthetic HA2 influenza fusion peptide analog with model membranes. AB - The interaction of the synthetic 21 amino acid peptide (AcE4K) with 1-oleoyl-2 [caproyl-7-NBD]-sn-glycero-3-phosphocholine membranes is used as a model system for the pH-sensitive binding of fusion peptides to membranes. The sequence of AcE4K (Ac-GLFEAIAGFIENGWEGMIDGK) is based on the sequence of the hemagglutinin HA2 fusion peptide and has similar partitioning into phosphatidylcholine membranes as the viral peptide. pH-dependent partitioning in the membrane, circular dichroism, tryptophan fluorescence, change of membrane area, and membrane strength, are measured to characterize various key aspects of the peptide-membrane interaction. The experimental results show that the partitioning of AcE4K in the membrane is pH dependent. The bound peptide inserts in the membrane, which increases the overall membrane area in a pH-dependent manner, however the depth of insertion of the peptide in the membrane is independent of pH. This result suggests that the binding of the peptide to the membrane is driven by the protonation of its three glutamatic acids and the aspartic acid, which results in an increase of the number of bound molecules as the pH decreases from pH 7 to 4.5. The transition between the bound state and the free state is characterized by the Gibbs energy for peptide binding. This Gibbs energy for pH 5 is equal to -30.2 kJ/mol (-7.2 kcal/mol). Most of the change of the Gibbs energy during the binding of AcE4K is due to the enthalpy of binding -27.3 kJ/mol (-6.5 kcal/mol), while the entropy change is relatively small and is on the order of 6.4 J/mol.K (2.3 cal/mol.K). The energy barrier separating the bound and the free state, is characterized by the Gibbs energy of the transition state for peptide adsorption. This Gibbs energy is equal to 51.3 kJ/mol (12.3 kcal/mol). The insertion of the peptide into the membrane is coupled with work for creation of a vacancy for the peptide in the membrane. This work is calculated from the measured area occupied by a single peptide molecule (220 A(2)) and the membrane elasticity (190 mN/m), and is equal to 15.5 kJ/mol (3.7 kcal/mol). The comparison of the work for creating a vacancy and the Gibbs energy of the transition state shows that the work for creating a vacancy may have significant effect on the rate of peptide insertion and therefore plays an important role in peptide binding. Because the work for creating a vacancy depends on membrane elasticity and the elasticity of the membrane is dependent on membrane composition, this provides a tool for modulating the pH for membrane instability by changing membrane composition. The insertion of the peptide in the membrane does not affect the membrane permeability for water, which shows that the peptide does not perturb substantially the packing of the hydrocarbon region. However, the ability of the membrane to retain solutes in the presence of peptide is compromised, suggesting that the inserted peptide promotes formation of short living pores. The integrity of the membrane is substantially compromised below pH 4.8 (threshold pH), when large pores are formed and the membrane breaks down. The binding of the peptide in the pore region is reversible, and the pore size varies on the experimental conditions, which suggests that the peptide in the pore region does not form oligomers. PMID- 11423415 TI - Two-dimensional infrared correlation spectroscopy study of the aggregation of cytochrome c in the presence of dimyristoylphosphatidylglycerol. AB - Two-dimensional infrared correlation spectroscopy (2D-IR) was used in this study to investigate the aggregation of cytochrome c in the presence of dimyristoylphosphatidylglycerol. The influence of temperature on the aggregation has been evaluated by monitoring the intensity of a band at 1616 cm(-1), which is characteristic of aggregated proteins, and the 2D-IR analysis has been used to determine the various secondary structure components of cytochrome c involved before and during its aggregation. The 2D-IR correlation analysis clearly reveals for the first time that aggregation starts to occur between nearly native proteins, which then unfold, yielding to further aggregation of the protein. Later in the aggregation process, the formation of intermolecular bonds and unfolding of the alpha-helices appear to be simultaneous. These results lead us to propose a two-step aggregation process. Finally, the results obtained during the heating period clearly indicate that before the protein starts to aggregate, there is a loosening of the tertiary structure of cytochrome c, resulting in a decrease of the beta-sheet content and an increase of the amount of beta-turns. This study clearly demonstrates the potential of 2D-IR spectroscopy to investigate the aggregation of proteins and this technique could therefore be applied to other proteins such as those involved in fibrilogenesis. PMID- 11423416 TI - Thermodynamic features of myosin filament suspensions: implications for the modeling of muscle contraction. AB - The analysis of myosin filament suspensions shows that these solutions are characterized by highly nonideal behavior. From these data a model is constructed that allows us to predict that 1) when subjected to an increasing protein osmotic pressure, myosin filaments experience an elastic deformation, which is not linearly related to the acting force; and 2) at constant protein osmotic pressure, when the cross-bridges of the myosin filaments are subjected to an external, nonosmotic force parallel to the filament axis, they are deformed and the water activity coefficient is altered. As a consequence, in muscle, passive and active shortening of the sarcomere is expected to promote the change of the water-water and of the water-protein interactions. We thus propose to depict muscle contraction as a chemo-osmoelastic transduction, where the analysis of the energy partition during the power stroke requires consideration of the osmotic factor in addition to the chemoelastic ones. PMID- 11423417 TI - Proximity relationships between residue 117 of rabbit skeletal troponin-I and residues in troponin-C and actin. AB - We used resonance energy transfer and site-directed photo-cross-linking to probe the Ca(2+)-dependent proximity relationships between residue 117 next to the C terminus of the inhibitory region in rabbit skeletal troponin-I (TnI) and residues in troponin-C (TnC) and in actin. A mutant TnI that contains a single cysteine at position 117 (I117) was constructed, and the distance between TnI residue 117 and TnC residue 98 was measured with the following results: for both the binary TnC-TnI complex and the ternary troponin complex, this distance was 30 and 41 A in the presence and absence of Ca(2+), respectively. The distance between TnI residue 117 and Cys374 of actin was 48 and 41 A in the presence and absence of Ca(2+), respectively. Six additional distances from this TnI residue to cysteines in TnC mutants were measured and used to localize this residue with respect to the crystal structure of TnC. The results show that in the presence of Ca(2+) it is localized near the B and C helices of TnC's N-terminal domain. In the absence of Ca(2+) this residue moves away from this location by approximately 8 A. Photo-cross-linking experiments show that I117 labeled with 4 maleimidobenzophenone photo-cross-linked to TnC but not to actin in both the presence and absence of Ca(2+). Taken together these results provide independent experimental support for the proposal (Y. Luo, J. L. Wu, B. Li, K. Langsetmo, J. Gergely, and T. Tao, 2000, J. Mol. Biol. 296:899-910) that upon Ca(2+) removal the region comprising TnI residues 114-125 triggers the movements of residues 89 113 and 130-150 toward actin, but does not itself interact with actin. PMID- 11423418 TI - Regulation of force development studied by photolysis of caged ADP in rabbit skinned psoas fibers. AB - The present study examined the effects of Ca(2+) and strongly bound cross-bridges on tension development induced by changes in the concentration of MgADP. Addition of MgADP to the bath increased isometric tension over a wide range of [Ca(2+)] in skinned fibers from rabbit psoas muscle. Tension-pCa (pCa is -log [Ca(2+)]) relationships and stiffness measurements indicated that MgADP increased mean force per cross-bridge at maximal Ca(2+) and increased recruitment of cross bridges at submaximal Ca(2+). Photolysis of caged ADP to cause a 0.5 mM MgADP jump initiated an increase in isometric tension under all conditions examined, even at pCa 6.4 where there was no active tension before ADP release. Tension increased monophasically with an observed rate constant, k(ADP), which was similar in rate and Ca(2+) sensitivity to the rate constant of tension re development, k(tr), measured in the same fibers by a release-re-stretch protocol. The amplitude of the caged ADP tension transient had a bell-shaped dependence on Ca(2+), reaching a maximum at intermediate Ca(2+) (pCa 6). The role of strong binding cross-bridges in the ADP response was tested by treatment of fibers with a strong binding derivative of myosin subfragment 1 (NEM-S1). In the presence of NEM-S1, the rate and amplitude of the caged ADP response were no longer sensitive to variations in the level of activator Ca(2+). The results are consistent with a model in which ADP-bound cross-bridges cooperatively activate the thin filament regulatory system at submaximal Ca(2+). This cooperative interaction influences both the magnitude and kinetics of force generation in skeletal muscle. PMID- 11423419 TI - Folding properties of functional domains of tropomodulin. AB - Tropomodulin (Tmod) stabilizes the actin-tropomyosin filament by capping the slow growing end (P-end). The N- and C-terminal halves play distinct roles; the N terminal half interacts with the N-terminal region of tropomyosin, whereas the C terminal half interacts with actin. Our previous study (A. Kostyukova, K. Maeda, E. Yamauchi, I. Krieger, and Y. Maeda Y., 2000, Eur. J. Biochem. 267:6470-6475) suggested that the two halves are also structurally distinct from each other. We have now studied the folding properties of the two halves, by circular dichroism spectroscopy and by differential scanning calorimetry of the expressed chicken E type tropomodulin and its large fragments. The results showed that the C-terminal half represents a single, independently folded unit that melts cooperatively through a two-state transition. In contrast, the N-terminal half lacks a definite tertiary structure in solution. The binding of N11, a fragment that corresponds to the first 91 amino acids of the tropomodulin, to tropomyosin substantially stabilized the tropomyosin. This may indicate that the flexible structure of the N-terminal half of tropomodulin in solution is required for binding to tropomyosin and that the N-terminal half acquires its tertiary structure upon binding to tropomyosin. PMID- 11423420 TI - Conformations of an adenine bulge in a DNA octamer and its influence on DNA structure from molecular dynamics simulations. AB - Molecular dynamics simulations have been applied to the DNA octamer d(GCGCA GAAC). d(GTTCGCGC), which has an adenine bulge at the center to determine the pathway for interconversion between the stacked and extended forms. These forms are known to be important in the molecular recognition of bulges. From a total of ~35 ns of simulation time with the most recent CHARMM27 force field a variety of distinct conformations and subconformations are found. Stacked and fully looped out forms are in excellent agreement with experimental data from NMR and x-ray crystallography. Furthermore, in a number of conformations the bulge base associates with the minor groove to varying degrees. Transitions between many of the conformations are observed in the simulations and used to propose a complete transition pathway between the stacked and fully extended conformations. The effect on the surrounding DNA sequence is investigated and biological implications of the accessible conformational space and the suggested transition pathway are discussed, in particular for the interaction of the MS2 replicase operator RNA with its coat protein. PMID- 11423421 TI - Sedimentation analysis of novel DNA structures formed by homo-oligonucleotides. AB - Sedimentation velocity analysis has been used to examine the base-specific structural conformations and unusual hydrogen bonding patterns of model oligonucleotides. Homo-oligonucleotides composed of 8-28 residues of dA, dT, or dC nucleotides in 100 mM sodium phosphate, pH 7.4, at 20 degrees C behave as extended monomers. Comparison of experimentally determined sedimentation coefficients with theoretical values calculated for assumed helical structures show that dT and dC oligonucleotides are more compact than dA oligonucleotides. For dA oligonucleotides, the average width (1.7 nm), assuming a cylindrical model, is smaller than for control duplex DNA whereas the average rise per base (0.34 nm) is similar to that of B-DNA. For dC and dT oligonucleotides, there is an increase in the average widths (1.8 nm and 2.1 nm, respectively) whereas the average rise per base is smaller (0.28 nm and 0.23 nm, respectively). A significant shape change is observed for oligo dC(28) at lower temperatures (10 degrees C), corresponding to a fourfold decrease in axial ratio. Optical density, circular dichroism, and differential scanning calorimetry data confirm this shape change, attributable from nuclear magnetic resonance analysis to i-motif formation. Sedimentation equilibrium studies of oligo dG(8) and dG(16) reveal extensive self-association and the formation of G-quadruplexes. Continuous distribution analysis of sedimentation velocity data for oligo dG(16) identifies the presence of discrete dimers, tetramers, and dodecamers. These studies distinguish the conformational and colligative properties of the individual bases in DNA and their inherent capacity to promote specific folding pathways. PMID- 11423422 TI - Photoinduced transient absorbance spectra of P840/P840(+) and the FMO protein in reaction centers of Chlorobium vibrioforme. AB - The kinetics of photoinduced absorbance changes in the 400-ns to 100-ms time range were studied between 770 and 1025 nm in reaction center core (RCC) complexes isolated from the green sulfur bacterium Chlorobium vibrioforme. A global, multiple stretched-exponential analysis shows the presence of two distinct but strongly overlapping spectra. The spectrum of the 70-micros component consists of a broad bleaching with two minima at 810 and 825 nm and a broad positive band at wavelengths greater than 865 nm and is assigned to the decay of (3)Bchl a of the Fenna-Matthews-Olson (FMO) protein. The contribution of the 70-micros component correlates with the amount of FMO protein in the isolated RCC complex. The spectrum of the 1.6-micros component has a sharp bleaching at 835 nm, a maximum at 805 nm, a broad positive band at wavelengths higher than 865 nm, and a broad negative band at wavelengths higher than 960 nm. When the RCC is incubated with inorganic iron and sulfur, the 1.6-micros component is replaced by a component with a lifetime of approximately 40 micros, consistent with the reconstruction of the F(X) cluster. We propose that the 1.6-micros component results from charge recombination between P840(+) and an intermediate electron acceptor operating between A(0) and F(X). Our studies in Chlorobium RCCs show that approaches that employ a single wavelength in the measurement of absorption changes have inherent limitations and that a global kinetic analysis at multiple wavelengths in the near-infrared is required to reliably separate absorption changes due to P840/P840(+) from the decay of (3)Bchl a in the FMO protein. PMID- 11423423 TI - Static and time-resolved step-scan Fourier transform infrared investigations of the photoreaction of halorhodopsin from Natronobacterium pharaonis: consequences for models of the anion translocation mechanism. AB - The molecular changes during the photoreaction of halorhodopsin from Natronobacterium pharaonis have been monitored by low-temperature static and by time-resolved step-scan Fourier transform infrared difference spectroscopy. In the low-temperature L spectrum anions only influence a band around 1650 cm(-1), tentatively assigned to the C=N stretch of the protonated Schiff base of L. The analysis of the time-resolved spectra allows to identify the four states: K, L(1), L(2), and O. Between L(1) and L(2), only the apoprotein undergoes alterations. The O state is characterized by an all-trans chromophore and by rather large amide I spectral changes. Because in our analysis the intermediate containing O is in equilibrium with a state indistinguishable from L(2), we are unable to identify an N-like state. At very high chloride concentrations (>5 M), we observe a branching of the photocycle from L(2) directly back to the dark state, and we provide evidence for direct back-isomerization from L(2). This branching leads to the reported reduction of transport activity at such high chloride concentrations. We interpret the L(1) to L(2) transition as an accessibility change of the anion from the extracellular to the cytosolic side, and the large amide I bands in O as an indication for opening of the cytosolic channel from the Schiff base toward the cytosolic surface and/or as indication for changes of the binding constant of the release site. PMID- 11423424 TI - Time-resolved fluorescence emission measurements of photosystem I particles of various cyanobacteria: a unified compartmental model. AB - Photosystem I (PS-I) contains a small fraction of chlorophylls (Chls) that absorb at wavelengths longer than the primary electron donor P700. The total number of these long wavelength Chls and their spectral distribution are strongly species dependent. In this contribution we present room temperature time-resolved fluorescence data of five PS-I core complexes that contain different amounts of these long wavelength Chls, i.e., monomeric and trimeric photosystem I particles of the cyanobacteria Synechocystis sp. PCC 6803, Synechococcus elongatus, and Spirulina platensis, which were obtained using a synchroscan streak camera. Global analysis of the data reveals considerable differences between the equilibration components (3.4-15 ps) and trapping components (23-50 ps) of the various PS-I complexes. We show that a relatively simple compartmental model can be used to reproduce all of the observed kinetics and demonstrate that the large kinetic differences are purely the result of differences in the long wavelength Chl content. This procedure not only offers rate constants of energy transfer between and of trapping from the compartments, but also well-defined room temperature emission spectra of the individual Chl pools. A pool of red shifted Chls absorbing around 702 nm and emitting around 712 nm was found to be a common feature of all studied PS-I particles. These red shifted Chls were found to be located neither very close to P700 nor very remote from P700. In Synechococcus trimeric and Spirulina monomeric PS-I cores, a second pool of red Chls was present which absorbs around 708 nm, and emits around 721 nm. In Spirulina trimeric PS-I cores an even more red shifted second pool of red Chls was found, absorbing around 715 nm and emitting at 730 nm. PMID- 11423425 TI - Regio-selective detection of dynamic structure of transmembrane alpha-helices as revealed from (13)C NMR spectra of [3-13C]Ala-labeled bacteriorhodopsin in the presence of Mn2+ ion. AB - 13C Nuclear magnetic resonance (NMR) spectra of [3-(13)C]Ala-labeled bacteriorhodopsin (bR) were edited to give rise to regio-selective signals from hydrophobic transmembrane alpha-helices by using NMR relaxation reagent, Mn(2+) ion. As a result of selective suppression of (13)C NMR signals from the surfaces in the presence of Mn(2+) ions, several (13)C NMR signals of Ala residues in the transmembrane alpha-helices were identified on the basis of site-directed mutagenesis without overlaps from (13)C NMR signals of residues located near the bilayer surfaces. The upper bound of the interatomic distances between (13)C nucleus in bR and Mn(2+) ions bound to the hydrophilic surface to cause suppressed peaks by the presence of Mn(2+) ion was estimated as 8.7 A to result in the signal broadening to 100 Hz and consistent with the data based on experimental finding. The Ala C(beta) (13)C NMR peaks corresponding to Ala-51, Ala-53, Ala-81, Ala-84, and Ala-215 located around the extracellular half of the proton channel and Ala-184 located at the kink in the helix F were successfully identified on the basis of (13)C NMR spectra of bR in the presence of Mn(2+) ion and site-directed replacement of Ala by Gly or Val. Utilizing these peaks as probes to observe local structure in the transmembrane alpha-helices, dynamic conformation of the extracellular half of bR at ambient temperature was examined, and the local structures of Ala-215 and 184 were compared with those elucidated at low temperature. Conformational changes in the transmembrane alpha-helices induced in D85N and E204Q and its long-range transmission from the proton release site to the site around the Schiff base in E204Q were also examined. PMID- 11423426 TI - Influence of the heme pocket conformation on the structure and vibrations of the Fe-CO bond in myoglobin: a QM/MM density functional study. AB - The influence of the distal pocket conformation on the structure and vibrations of the heme-CO bond in carbonmonoxy myoglobin (MbCO) is investigated by means of hybrid QM/MM calculations based on density functional theory combined with a classical force field. It is shown that the heme-CO structure (QM treated) is quite rigid and not influenced by the distal pocket conformation (MM treated). This excludes any relation between FeCO distortions and the different CO absorptions observed in the infrared spectra of MbCO (A states). In contrast, both the CO stretch frequency and the strength of the CO...His64 interaction are very dependent on the orientation and tautomerization state of His64. Our calculations indicate that the CO...N(epsilon) type of approach does not contribute to the A states, whereas the CO...H-N(epsilon) interaction is the origin of the A(1) and A(3) states, the His64 residue being protonated at N(epsilon). The strength of the CO...His64 interaction is quantified, in comparison with the analogous O(2)...His64 interaction and with the observed changes in the CO stretch frequency. Additional aspects of the CO...His64 interaction and its biological implications are discussed. PMID- 11423427 TI - The assembly of amyloidogenic yeast sup35 as assessed by scanning (atomic) force microscopy: an analogy to linear colloidal aggregation? AB - Amyloidosis is a class of diseases caused by protein aggregation and deposition in various tissues and organs. In this paper, a yeast amyloid-forming protein Sup35 was used as a model for understanding amyloid fiber formation. The dynamics of amyloid formation by Sup35 were studied with scanning force microscopy. We found that: 1) the assembly of Sup35 fibers begins with individual NM peptides that aggregate to form large beads or nucleation units which, in turn, form dimers, trimers, tetramers and longer linear assemblies appearing as a string of beads; 2) the morphology of the linear assemblies differ; and 3) fiber assembly suggests an analogy to the aggregation of colloidal particles. A dipole assembly model is proposed based on this analogy that will allow further experimental testing. PMID- 11423428 TI - Biophysical characterization of the influence of salt on tetrameric SecB. AB - SecB is a tetrameric chaperone, with a monomeric molecular mass of 17 kDa, that is involved in protein translocation in Escherichia coli. It has been hypothesized that SecB undergoes a conformational change as a function of the salt concentration. To gain more insight into the salt-dependent behavior of SecB, we studied the protein in solution by dynamic light scattering, size exclusion chromatography, analytical ultracentrifugation, and small angle neutron scattering. The results clearly demonstrate the large influence of the salt concentration on the behavior of SecB. At high salt concentration, SecB is a non spherical protein with a radius of gyration of 3.4 nm. At low salt concentration the hydrodynamic radius of the protein is apparently decreased, whereas the ratio of the frictional coefficients is increased. The protein solution behaves in a non-ideal way at low salt concentrations, as was shown by the analytical ultracentrifugation data and a pronounced interparticle effect observed by small angle neutron scattering. A possible explanation is a change in surface charge distribution dependent on the salt concentration in the solvent. We summarize our data in a model for the salt-dependent conformation of tetrameric SecB. PMID- 11423429 TI - Aplysia attractin: biophysical characterization and modeling of a water-borne pheromone. AB - Attractin, a 58-residue protein secreted by the mollusk Aplysia californica, stimulates sexually mature animals to approach egg cordons. Attractin from five different Aplysia species are approximately 40% identical in sequence. Recombinant attractin, expressed in insect cells and purified by reverse-phase high-performance liquid chromatography (RP-HPLC), is active in a bioassay using A. brasiliana; its circular dichroism (CD) spectrum indicates a predominantly alpha-helical structure. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) characterization of proteolytic fragments identified disulfide bonds between the six conserved cysteines (I-VI, II-V, III-IV, where the Roman numeral indicates the order of occurrence in the primary sequence). Attractin has no significant similarity to any other sequence in the database. The protozoan Euplotes pheromones were selected by fold recognition as possible templates. These diverse proteins have three alpha-helices, with six cysteine residues disulfide-bonded in a different pattern from attractin. Model structures with good stereochemical parameters were prepared using the EXDIS/DIAMOD/FANTOM program suite and constraints based on sequence alignments with the Euplotes templates and the attractin disulfide bonds. A potential receptor-binding site is suggested based on these data. Future structural characterization of attractin will be needed to confirm these models. PMID- 11423430 TI - HIV-1 integrase catalytic core: molecular dynamics and simulated fluorescence decays. AB - Two molecular dynamics simulations have been carried out on the HIV-1 integrase catalytic core starting from fully determined crystal structures. During the first one, performed in the absence of divalent cation (6-ns long), the catalytic core took on two main conformations. The conformational transition occurs at approximately 3.4 ns. In contrast, during the second one, in the presence of Mg(2+) (4-ns long), there were no such changes. The molecular dynamics simulations were used to compute the fluorescence intensity decays emitted by the four tryptophan residues considered as the only chromophores. The decay was computed by following, frame by frame, the amount of chromophores that remained excited at a certain time after light absorption. The simulation took into account the quenching through electron transfer to the peptide bond and the fluorescence resonance energy transfer between the chromophores. The fit to the experimental intensity decays obtained at 5 degrees C and at 30 degrees C is very good. The fluorescence anisotropy decays were also simulated. Interestingly, the fit to the experimental anisotropy decay was excellent at 5 degrees C and rather poor at 30 degrees C. Various hypotheses such as dimerization and abnormal increase of uncorrelated internal motions are discussed. PMID- 11423431 TI - Structure and hydration of the DNA-human topoisomerase I covalent complex. AB - The structure and hydration of reconstituted human topoisomerase I comprising the core and the carboxyl-terminal domains in covalent complex with 22-basepair DNA duplex has been investigated by molecular dynamics simulation. The structure and the intermolecular interactions were found to be well maintained over the simulation. The complex displays a high degree of flexibility of the contact area, confirmed by the presence of numerous water-mediated protein-DNA hydrogen bonds comparable in quantity and distribution to the direct ones. The interaction between the enzyme and the solvent also provides the key for interpreting the experimental reduction of activity or affinity observed upon single residue mutation. Finally, four long lasting water molecules are observed in the proximity of the active site, one of which in the appropriate position to accept a proton from the active Tyr723. PMID- 11423432 TI - Development of a time-resolved fluorometric method for observing hybridization in living cells using fluorescence resonance energy transfer. AB - We previously showed that a specific kind of mRNA (c-fos) was detected in a living cell under a microscope by introducing two fluorescently labeled oligodeoxynucleotides, each labeled with donor or acceptor, into the cytoplasm, making them hybridize to adjacent locations on c-fos mRNA, and taking images of fluorescence resonance energy transfer (FRET) (A. Tsuji, H. Koshimoto, Y. Sato, M. Hirano. Y. Sei-Iida, S. Kondo, and K. Ishibashi, 2000, Biophys. J. 78:3260 3274). On the formed hybrid, the distance between donor and acceptor becomes close and FRET occurs. To observe small numbers of mRNA in living cells using this method, it is required that FRET fluorescence of hybrid must be distinguished from fluorescence of excess amounts of non-hybridizing probes and from cell autofluorescence. To meet these requirements, we developed a time resolved method using acceptor fluorescence decays. When a combination of a donor having longer fluorescence lifetime and an acceptor having shorter lifetime is used, the measured fluorescence decays of acceptors under FRET becomes slower than the acceptor fluorescence decay with direct excitation. A combination of Bodipy493/503 and Cy5 was selected as donor and acceptor. When the formed hybrid had a configuration where the target RNA has no single-strand part between the two fluorophores, the acceptor fluorescence of hybrid had a sufficiently longer delay to detect fluorescence of hybrid in the presence of excess amounts of non hybridizing probes. Spatial separation of 10-12 bases between two fluorophores on the hybrid is also required. The decay is also much slower than cell autofluorescence, and smaller numbers of hybrid were detected with less interference of cell autofluorescence in the cytoplasm of living cells under a time-resolved fluorescence microscope with a time-gated function equipped camera. The present method will be useful when observing induced expressions of mRNA in living cells. PMID- 11423433 TI - Electron paramagnetic resonance evidence for binding of Cu(2+) to the C-terminal domain of the murine prion protein. AB - Transmissible spongiform encephalopathies in mammals are believed to be caused by scrapie form of prion protein (PrP(Sc)), an abnormal, oligomeric isoform of the monomeric cellular prion protein (PrP(C)). One of the proposed functions of PrP(C) in vivo is a Cu(II) binding activity. Previous studies revealed that Cu(2+) binds to the unstructured N-terminal PrP(C) segment (residues 23-120) through conserved histidine residues. Here we analyzed the Cu(II) binding properties of full-length murine PrP(C) (mPrP), of its isolated C-terminal domain mPrP(121-231) and of the N-terminal fragment mPrP(58-91) in the range of pH 3-8 with electron paramagnetic resonance spectroscopy. We find that the C-terminal domain, both in its isolated form and in the context of the full-length protein, is capable of interacting with Cu(2+). Three Cu(II) coordination types are observed for the C-terminal domain. The N-terminal segment mPrP(58-91) binds Cu(2+) only at pH values above 5.0, whereas both mPrP(121-231) and mPrP(23-231) already show identical Cu(II) coordination in the pH range 3-5. As the Cu(2+) binding N-terminal segment 58-91 is not required for prion propagation, our results open the possibility that Cu(2+) ions bound to the C-terminal domain are involved in the replication of prions, and provide the basis for further analytical studies on the specificity of Cu(II) binding by PrP. PMID- 11423434 TI - High-Q dynamic force microscopy in liquid and its application to living cells. AB - We present a new dynamic force microscopy technique for imaging in liquids in the piconewton regime. The low quality factor (Q) of the cantilever is increased up to three orders of magnitude by the implementation of a positive feedback control. The technique also includes a phase-locked loop unit to track the resonance of the cantilever. Experiments and computer simulations indicate that the tip-sample forces are below 100 pN, about two orders of magnitude lower than in conventional tapping mode atomic force microscopy. Furthermore, the spectroscopic ability is greatly enhanced. Either the phase shift or the resonant frequency shows a high sensitivity to variations in either the energy dissipation or conservative interactions between the tip and the sample, respectively. The potential of this technique is demonstrated by imaging living cells. PMID- 11423435 TI - Fluorescence lifetime heterogeneity in aggregates of LHCII revealed by time resolved microscopy. AB - Two-photon excitation, time-resolved fluorescence microscopy was used to investigate the fluorescence quenching mechanisms in aggregates of light harvesting chlorophyll a/b pigment protein complexes of photosystem II from green plants (LHCII). Time-gated microscopy images show the presence of large heterogeneity in fluorescence lifetimes not only for different LHCII aggregates, but also within a single aggregate. Thus, the fluorescence decay traces obtained from macroscopic measurements reflect an average over a large distribution of local fluorescence kinetics. This opens the possibility to resolve spatially different structural/functional units in chloroplasts and other heterogeneous photosynthetic systems in vivo, and gives the opportunity to investigate individually the excited states dynamics of each unit. We show that the lifetime distribution is sensitive to the concentration of quenchers contained in the system. Triplets, which are generated at high pulse repetition rates of excitation (>1 MHz), preferentially quench domains with initially shorter fluorescence lifetimes. This proves our previous prediction from singlet-singlet annihilation investigations (Barzda, V., V. Gulbinas, R. Kananavicius, V. Cervinskas, H. van Amerongen, R. van Grondelle, and L. Valkunas. 2001. Biophys. J. 80:2409-2421) that shorter fluorescence lifetimes originate from larger domains in LHCII aggregates. We found that singlet-singlet annihilation has a strong effect in time-resolved fluorescence microscopy of connective systems and has to be taken into consideration. Despite that, clear differences in fluorescence decays can be detected that can also qualitatively be understood. PMID- 11423436 TI - Domain formation in models of the renal brush border membrane outer leaflet. AB - The plasma membrane outer leaflet plays a key role in determining the existence of rafts and detergent-resistant membrane domains. Monolayers with lipid composition mimicking that of the outer leaflet of renal brush border membranes (BBM) have been deposited on mica and studied by atomic force microscopy. Sphingomyelin (SM) and palmitoyloleoyl phosphatidylcholine (POPC) mixtures, at molar ratios varying from 2:1 to 4:1, were phase-separated into liquid condensed (LC) SM-enriched phase and liquid expanded (LE) POPC-enriched phase. The LC phase accounted for 33 and 58% of the monolayers surface for 2:1 and 4:1 mixtures, respectively. Addition of 20-50 mol % cholesterol (Chl) to the SM/POPC (3:1) mixtures induced marked changes in the topology of monolayers. Whereas Chl promoted the connection between SM domains at 20 mol %, increasing Chl concentration progressively reduced the size of domains and the height differences between the phases. Lateral heterogeneity was, however, still present at 33 mol % Chl. The results indicate that the lipid composition of the outer leaflet is most likely responsible for the BBM thermotropic transition properties. They also strongly suggest that the common maneuver that consists of depleting membrane cholesterol to suppress rafts does not abolish the lateral heterogeneity of BBM membranes. PMID- 11423437 TI - Single molecule spectroscopy on the light-harvesting complex II of higher plants. AB - Spectroscopic and polarization properties of single light-harvesting complexes of higher plants (LHC-II) were studied at both room temperature and T < 5 K. Monomeric complexes emit roughly linearly polarized fluorescence light thus indicating the existence of only one emitting state. Most probably this observation is explained by efficient triplet quenching restricted to one chlorophyll a (Chl a) molecule or by rather irreversible energy transfer within the pool of Chl a molecules. LHC-II complexes in the trimeric (native) arrangement bleach in a number of steps, suggesting localization of excitations within the monomeric subunits. Interpretation of the fluorescence polarization properties of trimers requires the assumption of transition dipole moments tilted out of the symmetry plane of the complex. Low-temperature fluorescence emission of trimers is characterized by several narrow spectral lines. Even at lowest excitation intensities, we observed considerable spectral diffusion most probably due to low temperature protein dynamics. These results also indicate weak interaction between Chls belonging to different monomeric subunits within the trimer thus leading to a localization of excitations within the monomer. The experimental results demonstrate the feasibility of polarization sensitive studies on single LHC-II complexes and suggest an application for determination of the Chl transition-dipole moment orientations, a key issue in understanding the structure-function relationships. PMID- 11423438 TI - Design of functionalized lipids and evidence for their binding to photosystem II core complex by oxygen evolution measurements, atomic force microscopy, and scanning near-field optical microscopy. AB - Photosystem II core complex (PSII CC) absorbs light energy and triggers a series of electron transfer reactions by oxidizing water while producing molecular oxygen. Synthetic lipids with different alkyl chains and spacer lengths bearing functionalized headgroups were specifically designed to bind the Q(B) site and to anchor this large photosynthetic complex (240 kDa) in order to attempt two dimensional crystallization. Among the series of different compounds that have been tested, oxygen evolution measurements have shown that dichlorophenyl urea (DCPU) binds very efficiently to the Q(B) site of PSII CC, and therefore, that moiety has been linked covalently to the headgroup of synthetic lipids. The analysis of the monolayer behavior of these DCPU-lipids has allowed us to select ones bearing long spacers for the anchoring of PSII CC. Oxygen evolution measurements demonstrated that these long-spacer DCPU-lipids specifically bind to PSII CC and inhibit electron transfer. With the use of atomic force microscopy (AFM) and scanning near-field optical microscopy (SNOM), it was possible to visualize domains of PSII CC bound to DCPU-lipid monolayers. SNOM imaging has enabled us to confirm that domains observed by AFM were composed of PSII CC. Indeed, the SNOM topography images presented similar domains as those observed by AFM, but in addition, it allowed us to determine that these domains are fluorescent. Electron microscopy of these domains, however, has shown that the bound PSII CC was not crystalline. PMID- 11423439 TI - Synchrotron X-ray study of lung surfactant-specific protein SP-B in lipid monolayers. AB - This work reports the first x-ray scattering measurements to determine the effects of SP-B(1-25), the N-terminus peptide of lung surfactant-specific protein SP-B, on the structure of palmitic acid (PA) monolayers. In-plane diffraction shows that the peptide fluidizes a portion of the monolayer but does not affect the packing of the residual ordered phase. This implies that the peptide resides in the disordered phase, and that the ordered phase is essentially pure lipid, in agreement with fluorescence microscopy studies. X-ray reflectivity shows that the peptide is oriented in the lipid monolayer at an angle of approximately 56 degrees relative to the interface normal, with one end protruding past the hydrophilic region into the fluid subphase and the other end embedded in the hydrophobic region of the monolayer. The quantitative insights afforded by this study lead to a better understanding of the lipid/protein interactions found in lung surfactant systems. PMID- 11423440 TI - HIV-1 capsid protein forms spherical (immature-like) and tubular (mature-like) particles in vitro: structure switching by pH-induced conformational changes. AB - The viral genome and replicative enzymes of the human immunodeficiency virus are encased in a shell consisting of assembled mature capsid protein (CA). The core shell is a stable, effective protective barrier, but is also poised for dissolution on cue to allow transmission of the viral genome into its new host. In this study, static light scattering (SLS) and dynamic light scattering (DLS) were used to examine the entire range of the CA protein response to an environmental cue (pH). The CA protein assembled tubular structures as previously reported but also was capable of assembling spheres, depending on the pH of the protein solution. The switch from formation of one to the other occurred within a very narrow physiological pH range (i.e., pH 7.0 to pH 6.8). Below this range, only dimers were detected. Above this range, the previously described tubular structures were detected. The ability of the CA protein to form a spherical structure that is detectable by DLS but not by electron microscopy indicates that some assemblages are inherently sensitive to perturbation. The dimers in equilibrium with these assemblages exhibited distinct conformations: Dimers in equilibrium with the spherical form exhibited a compact conformation. Dimers in equilibrium with the rod-like form had an extended conformation. Thus, the CA protein possesses the inherent ability to form metastable structures, the morphology of which is regulated by an environmentally-sensitive molecular switch. Such metastable structures may exist as transient intermediates during the assembly and/or disassembly of the virus core. PMID- 11423441 TI - Factoring cost, is phacoemulsification still the procedure of choice? PMID- 11423442 TI - HAART, CMV retinitis, and monitoring. PMID- 11423443 TI - Alternative eye care. PMID- 11423444 TI - Ophthalmologists and optometrists--interesting times? PMID- 11423445 TI - Blindness in the eastern Mediterranean countries. PMID- 11423446 TI - Cataract blindness and barriers to uptake of cataract surgery in a rural community of northern Nigeria. AB - AIMS: A population based cross sectional survey was conducted to determine the magnitude of cataract blindness and the barriers to uptake of cataract services in a rural community of northern Nigeria. METHODS: 1461 people out of 1924 registered eligible people were examined. The study population was chosen by two stage cluster random sampling. In the first sampling stage 15 villages were randomly chosen while in the final stage 170 people who were 40 years and over were selected in each village. Each selected person had visual acuity recorded for both eyes. Those with vision of less than 3/60 in the better eye were assessed for cataract. People with cataract were asked why they had not sought medical attention. RESULTS: A blindness prevalence of 8.2% (95% CI 5.8%-10.5%) was found among the sampled population. Cataract was responsible for 44.2% of the blindness. Thus, a cataract blindness prevalence of 3.6% was found. The cataract surgical coverage (people) was 4.0% and the couching coverage (people) was 18%. The main barrier to seeking cataract surgery was cost of the service (61%). CONCLUSION: Some regions of the world still have high burden of cataract blindness that needs attention. Such areas need an effective free cataract outreach programme. PMID- 11423447 TI - Human papillomavirus in pterygium. AB - AIM: To determine the prevalence and significance of human papillomaviral types in conjunctival pterygia. METHODS: Polymerase chain reaction technology was used to identify the presence of human papillomavirus (HPV) in 10 formalin fixed paraffin embedded pterygia samples. 10 conjunctival papillomas were used as positive controls. 20 conjunctival samples, 10 with primary acquired melanosis and 10 with malignant melanoma, were used as negative controls. Sample subgroups were of equal sex, race, and age distribution to eliminate bias. All samples were further analysed (for 21 HPV types) using dot-blot hybridisation techniques. RESULTS: HPV was identified in 90% of the conjunctival papillomas, 50% of the pterygia samples, but no HPV was detected in the negative control group. Two pterygia showed type 6, two type 11, and one type 16. These three HPV types were also detected in papillomas. CONCLUSION: These results suggest that HPV may be involved in the pathogenesis of pterygia and that broadly the same HPV types are found in pterygia and in papillomas. Persistent conjunctival HPV may possibly play a part in the recurrence of pterygia post excision but further larger studies are required to elucidate this hypothesis. PMID- 11423448 TI - Human papillomavirus in conjunctival papilloma. AB - AIM: To examine conjunctival papillomas for the presence of human papillomavirus (HPV) and koilocytosis. METHODS: Archival paraffin embedded tissue from 55 conjunctival papillomas was analysed for the presence of HPV by polymerase chain reaction and subsequent filter hybridisation. Histological sections of the 55 papillomas were evaluated for the presence of koilocytosis. RESULTS: HPV was present in 48 of 52 (92%) beta globin positive papillomas. HPV type 6/11 were found in 40 of 47 investigated papillomas and a double infection with HPV 6/11 and 16 was identified in a single papilloma. In six papillomas the HPV type could not be identified. Koilocytosis was present in 22 of 55 papillomas (40%). CONCLUSION: There is a strong association between HPV and conjunctival papillomas. HPV type 6/11 is the most common HPV type in conjunctival papilloma. The sensitivity of koilocytosis as an indicator of HPV in conjunctival papilloma is low. PMID- 11423449 TI - Deep lamellar keratoplasty with lyophilised tissue in the management of keratoconus. AB - AIMS: Data are presented on the use of deep lamellar keratoplasty (DLK) using lyophilised donor corneal tissue, in the management of patients with keratoconus (KC). METHOD: The results of DLK on 44 eyes (42 patients) are reported. The mean patient age was 29.8 years (range 10-56). Mean follow up was 25 months (range 6 100). In seven patients with mental handicap or severe mental illness, the collection of acuity and refractive data was limited. RESULTS: Perforation of Descemet's membrane (DM) occurred in nine cases (20%). A double anterior chamber formed in five cases, which resolved spontaneously in three patients. Persistent epithelial defects occurred in two cases, one of which necessitated replacement of the graft. The median postoperative uncorrected visual acuity was 6/36. The median corrected postoperative acuity was 6/9. Those with more than 1 year of follow up (n=25) had a significantly better acuity (p=0.015). This group achieved 6/12 or better in 80% (n=20) and 6/6 or better in 40% (n=10). The mean postoperative spherical error was +0.28 (SD 3.49) dioptres (D). The mean refractive cylinder was 3.85 (1.87) D. CONCLUSION: This detailed retrospective study of DLK for the treatment of patients with KC, with an average follow up of 2 years, highlights the advantages and disadvantages of this technique. PMID- 11423450 TI - Central corneal thickness measurements in patients with normal tension glaucoma, primary open angle glaucoma, pseudoexfoliation glaucoma, or ocular hypertension. AB - BACKGROUND/AIMS: Recent studies have revealed patients with ocular hypertension to have thicker than normal central corneas and those with normal tension glaucoma to have thinner than normal ones, as determined by ultrasonic pachymetry. Since corneal thickness measurements and applanation tonometric estimates of intraocular pressure (IOP) correlate positively, monitoring of the former parameter have served as the basis for adjusting readings pertaining to the latter, with the consequence that many patients have had to be reclassified. With a view to validating these pachymetric studies, the central corneal thickness was determined in patients with normal tension glaucoma, primary open angle glaucoma, pseudoexfoliation glaucoma, or ocular hypertension, as well as that of normal subjects, using optical low coherence reflectometry, which is a new and more precise method than ultrasonic pachymetry. METHODS: 34 patients with normal tension glaucoma, 20 with primary open angle glaucoma, 13 with pseudoexfoliation glaucoma, and 12 with ocular hypertension, together with 21 control subjects, were included in this observational, concurrent case-control study. One eye per individual was randomly selected for investigation. IOP was measured by Goldmann applanation tonometry and central corneal thickness by optical low coherence reflectometry. RESULTS: Central corneal thickness was significantly higher (p < or =0.001) in patients with ocular hypertension than in normal individuals or in subjects with either normal tension glaucoma, primary open angle glaucoma, or pseudoexfoliation glaucoma, there being no significant differences between the latter four groups. Patients with ocular hypertension were also significantly younger (p < or =0.003) than those within any of the three glaucomatous groups. CONCLUSION: This study confirms that a significant number of patients with ocular hypertension have normal IOPs after the appropriate adjustments have been made for deviations from normal in their central corneal thickness. The accurate measurement of this latter parameter is important not only for individual patient care, in permitting more precise estimations of IOP, but also for clinical studies, in assuring a more reliable classification of subjects. PMID- 11423451 TI - Intraocular pressure and pulsatile ocular blood flow after retrobulbar and peribulbar anaesthesia. AB - AIMS: This study investigated the effect of peribulbar and retrobulbar local anaesthesia on intraocular pressure (IOP) and pulsatile ocular blood flow (POBF), as such anaesthetic techniques may adversely affect these parameters. METHODS: 20 eyes of 20 patients who were to undergo phacoemulsification cataract surgery were prospectively randomised to receive peribulbar or retrobulbar anaesthesia. The OBF tonometer (OBF Labs, Wiltshire, UK) was used to simultaneously measure IOP and POBF before anaesthesia and 1 minute and 10 minutes after anaesthesia. Between group comparisons of age, baseline IOP, and baseline POBF were performed using the non-parametric Mann-Whitney test. Within group comparisons of IOP and POBF measured preanaesthesia and post-anaesthesia were performed using the non parametric Wilcoxon signed ranks test for both groups. RESULTS: There was no statistically significant IOP increase post-anaesthesia in either group. In the group receiving peribulbar anaesthesia, there was a significant reduction in POBF initially post-anaesthesia which recovered after 10 minutes. In the group receiving retrobulbar anaesthesia, there was a persistent statistically significant reduction in POBF. CONCLUSIONS: Retrobulbar and peribulbar injections have little effect on IOP. Ocular compression is not needed for IOP reduction when using local anaesthesia for cataract surgery. Conversely, POBF falls, at least for a short time, when anaesthesia for ophthalmic surgery is administered via a retrobulbar route or a peribulbar route. This reduction may be mediated by pharmacologically altered orbital vascular tone. It may be safer to use other anaesthetic techniques in patients with ocular vascular compromise. PMID- 11423452 TI - Effect of digital image compression on screening for diabetic retinopathy. AB - BACKGROUND/AIMS: Digital imaging is widely used for diabetic retinopathy screening. The storage and transmission of digital images can be facilitated by image compression. The authors aimed to assess the effect of image compression on the accuracy of grading diabetic retinopathy. METHODS: Forty nine 35 mm transparencies (17 with no retinopathy, eight with background, five with preproliferative, and 19 with proliferative retinopathy) were digitised and subjected to JPEG compression by 90%, 80%, 70%, and 0%. The 196 images were randomised and graded on a portable computer. Two masked graders assessed the images for grade of retinopathy and image quality (0-10). The sensitivity and specificity of retinopathy grading were calculated with a weighted kappa for grading agreement between levels of compression. RESULTS: The sensitivity of retinopathy grading was reduced by JPEG compression. At 90%, 80%, 70%, and 0% compression the sensitivities were 0.38, 0.50, 0.65, and 0.72, respectively; the specificity results were 1.00, 1.00, 0.83, and 0.84, respectively; and the weighted kappa scores were 0.60, 0.75, 0.77, and 0.84, respectively. The quality scores for 90%, 80%, 70%, 0% compression were 2.9 (SD 1.1, 95% CI; 2.7-3.2), 4.6 (SD 1.1, 95% CI; 3.0-5.6), 5.8 (SD1.5, 95% CI 5.0-6.6), 6.3 (SD1.4, 95% CI; 5.4 7.2) (p<0.01 for each intergroup comparison). CONCLUSION: The results demonstrate significant loss of sensitivity to the features of diabetic retinopathy with JPEG compression; this was compounded by the thin film transistor (TFT) screen. The authors found the quality of uncompressed images on TFT screens too poor to give grading sensitivities which reach current guidelines for diabetic retinopathy screening. PMID- 11423453 TI - Hyperhomocyst(e)inaemia, but not MTHFR C677T mutation, as a risk factor for non arteritic ischaemic optic neuropathy. AB - BACKGROUND/AIMS: Hyperhomocyst(e)inaemia has been identified as a strong risk factor for stroke, myocardial infarction, and deep vein thrombosis. A point mutation of methylene tetrahydrofolate reductase (MTHFR C677T) has been associated with increased plasma homocyst(e)ine levels. To investigate whether hyperhomocyst(e)inaemia and/or MTHFR C677T mutation are associated with non arteritic ischaemic optic neuropathy (NAION), a case-control study including 59 consecutive patients with NAION and 59 controls matched for age and sex was performed. METHODS: Fasting plasma homocyst(e)ine levels, MTHFR C677T genotypes, and plasma levels of folate and vitamin B-12 were determined. RESULTS: Mean plasma homocyst(e)ine levels were significantly higher in patients than in controls (11.8 (SD 5.7) micromol/l v 9.8 (2.5) micromol/l, p = 0.02). The odds ratio for patients with homocyst(e)ine levels exceeding the 95th percentile of control homocyst(e)ine levels was 5.8 (95% CI 1.5-21.4). Mean plasma folate levels were significantly lower in patients than in controls (4.3 (1.7) ng/ml v 5.5 (1.9) ng/ml, p = 0.001), whereas plasma vitamin B-12 levels did not differ significantly. Prevalence of the MTHFR C677T mutation was not significantly increased in patients with NAION compared with controls. CONCLUSION: These results suggest that hyperhomocyst(e)inaemia, but not MTHFR C677T mutation is associated with NAION. Determination of plasma homocyst(e)ine levels might be of diagnostic value in patients with NAION. PMID- 11423454 TI - Regionalisation of retinopathy of prematurity (ROP) screening improves compliance with guidelines: an audit of ROP screening in the Northern Region of England. AB - AIMS: This project was designed to determine whether a coordinated regional strategy can improve the implementation of national guidelines for screening for retinopathy of prematurity (ROP), and to identify causes for failure of compliance. METHODS: Retrospective case note audit relating to two periods, 1990 1 and 1994, involving all 17 neonatal intensive care units in the Northern Region of England. Between the two periods, a regional strategy was instituted in an endeavour to improve compliance. Babies born in or admitted to the units during the study periods who were eligible for ROP screening were included. Screening performance was assessed against a standard of 100% compliance with the guidelines. In the second audit period compliance with subsidiary standards was also measured, and reasons for failure were identified. RESULTS: Compliance improved from 47% (262/558) in the first audit cycle to 73% (264/360) in the second. Subgroup analysis in this second cycle indicated better compliance (93.3%) in higher risk babies (< or = 29 weeks' gestational age). Babies transferred between units, discharged home before screening, or who failed to qualify for screening on one of the two defined criteria, were more likely to be missed. CONCLUSION: A carefully implemented regional approach to screening resulted in a higher uptake for babies most at risk. Simple recommendations are made to achieve further improvement in compliance with the guidelines. The wider implications for screening in other conditions and in other areas and specialties are highlighted. PMID- 11423455 TI - Tritan colour contrast sensitivity function in refractive multifocal intraocular lenses. AB - AIMS: To compare tritan colour contrast sensitivity (CCS), without and with glare, in patients with refractive multifocal intraocular lenses (IOLs) and with monofocal intraocular lenses. METHODS: Tritan CCS was determined (Moorfields Vision System, CH Electronics) in 15 eyes (14 patients, 75.7 (+/-6.6) years) with a refractive multifocal IOL (Allergan SA 40N) and in 11 eyes (10 patients, 73.7 (+/-6.4) years) with a monofocal IOL (Allergan SI 40 NB). Measurements were made monocularly under mesopic conditions at a distance of 2 metres from the monitor with best distance refraction plus 0.5 D at 0.5, 1, 3, 6, 11.4, and 22.8 cycles per degree (cpd). The test was then repeated for the multifocal IOLs, adding minus 2.5 D to the best distance refraction to force the patient to use the near focus. Both lenses were also investigated under glare conditions with the same set-up and using the brightness acuity tester (BAT). RESULTS: The tritan CCS function without glare in multifocal lenses through the distance focus was nearly identical to that through the near focus. The following statistically significant differences were measured: the CCS function without glare for the multifocal lens was worse at 0.5 cpd and 1.0 cpd than that of the monofocal lens. In CCS testing of the multifocal group with glare at 6 cpd, the results through the distance focus were better than the results through the near focus. For the CCS function with glare, the values for the distance focus in the multifocal lens were worse than the values for the monofocal lens at 0.5 cpd and 1 cpd. In CCS testing with glare through the near focus and CCS testing through the monofocal lens, the monofocal lens performed better at 0.5 cpd, 1 cpd, 3 cpd, and 6 cpd. CONCLUSION: Refractive multifocal intraocular lenses influence tritan CCS function compared to monofocal lenses. PMID- 11423456 TI - Halo size under distance and near conditions in refractive multifocal intraocular lenses. AB - AIMS: To calculate the diameter of halos perceived by patients with multifocal intraocular lenses (IOLs) and to stimulate halos in patients with refractive multifocal IOLs in a clinical experiment. METHODS: Calculations were done to show the diameter of halos in the case of the bifocal intraocular lens. 24 patients with a refractive multifocal IOLs and five patients with a monofocal IOL were asked about their subjective observation of halos and were included in a clinical experiment using a computer program (Glare & Halo, FW Fitzke and C Lohmann, Tomey AG) which simulates a light source of 0.15 square degrees (sq deg) in order to stimulate and measure halos. Halo testing took place monoculary, under mesopic conditions through the distance and the near focus of the multifocal lens and through the focus of the monofocal lens. RESULTS: The halo diameter depends on the pupil diameter, the refractive power of the cornea, and distance focus of the multifocal IOL as well as the additional lens power for the near focus. 23 out of 24 patients with a refractive multifocal IOL described halos at night when looking at a bright light source. Only one patient was disturbed by the appearance of halos. Under test conditions, halos were detected in all patients with a refractive multifocal IOL. The halo area testing through the distance focus was 1.05 sq deg +/- 0.41, through the near focus 1.07 sq deg +/- 0.49 and in the monofocal lens 0.26 sq deg +/- 0.13. CONCLUSIONS: Under high contrast conditions halos can be stimulated in all patients with multifocal intraocular lenses. The halo size using the distance or the near focus is identical. PMID- 11423457 TI - Extracapsular cataract extraction compared with small incision surgery by phacoemulsification: a randomised trial. AB - BACKGROUND: Cataract extraction constitutes the largest surgical workload in ophthalmic units throughout the world. Extracapsular cataract extraction (ECCE), through a large incision, with insertion of an intraocular lens has been the most widely used method from 1982 until recently. Technological advances have led to the increasing use of phacoemulsification (Phako) to emulsify and remove the lens The technique requires a smaller incision, but requires substantial capital investment in theatre equipment. In this randomised trial we assessed the clinical outcomes and carried out an economic evaluation of the two procedures. METHODS: In this two centre randomised trial, 232 patients with age related cataract received ECCE, and 244 received small incision surgery by Phako. The main comparative outcomes were visual acuity, refraction, and complication rates. Resource use was monitored in the two trial centres and in an independent comparator centre. Costs calculated included average cost per procedure, at each stage of follow up. RESULTS: Phako was found to be clinically superior. Surgical complications and capsule opacity within 1 year after surgery were significantly less frequent, and a higher proportion achieved an unaided visual acuity of 6/9 or better (<0.2 logMAR) in the Phako group. Postoperative astigmatism was more stable in Phako. The average cost of a cataract operation and postoperative care within the trial was similar for the two procedures. With the input of additional spectacles for corrected vision at 6 months after surgery, the average cost per procedure was pound359.89 for Phako and pound367.57 for ECCE. CONCLUSION: Phako is clinically superior to ECCE and is cost effective. PMID- 11423458 TI - Interlenticular opacification in piggyback AcrySof intraocular lenses: explantation technique and laboratory investigations. AB - BACKGROUND/AIMS: Interlenticular opacification (ILO) is a recognised complication of piggyback intraocular lenses (IOLs). The aetiology, histopathology, and treatment are not clearly defined, however. METHODS: Two pairs of AcrySof IOLs were explanted from a patient with bilateral ILO. The explantation technique and surgical challenges of IOL exchanges are described. The explanted IOL complexes and a sample of the anterior capsule were examined by phase, polarising, and immunofluorescence microscopy. RESULTS: A 50 year old man developed ILO bilaterally after piggyback AcrySof IOL implantation. A central contact zone was surrounded by a homogeneous paracentral opacity possibly consisting of extracellular matrix previously laid down by proliferating lens epithelial cells (LECs). These opacities were in turn surrounded by interlenticular Elschnig pearl type opacities contiguous with the same material filling the periphery of the capsular bag. The IOL complexes were very adherent to the capsular bag and they had to be separated with the help of high viscosity viscoelastic before a single one piece PMMA IOL implantation via large limbal incisions. The sample of anterior capsule showed a ridge configuration from the piling of LECs in the site of apposition with the anterior capsule and cells showing different characteristics on either side of the ridge. CONCLUSION: Cellular proliferation, deposition of ECM from proliferating LECs, and capsular changes induced by cell metaplasia may lead to ILO formation in piggyback AcrySof IOLs. Careful separation of the AcrySof IOL complex from the capsule, meticulous clean up of the proliferating material, and implantation of single or dual in the bag PMMA IOLs through a large incision with capsulorrhexis enlargement may help in the prevention of recurrence of interface opacification. PMID- 11423459 TI - CMVR diagnoses and progression of CD4 cell counts and HIV viral load measurements in HIV patients on HAART. AB - AIM: To assess the impact of highly active antiretroviral therapy (HAART) on the prevalence and progression of CMV retinitis (CMVR) among AIDS patients with baseline CD4 cell counts <100 cells x 10(6)/l. METHODS: A longitudinal cohort study of 1292 patients. CD4 cell counts and HIV viral load measurements were obtained before commencing therapy, at 3 months, 1 year, 2 years, and at last follow up. The CMVR prevalence rate was measured for the subgroup with baseline CD4 cell counts <100 cells x 10(6)/l. CMVR adverse event (AE) rates per 100 person days at risk were calculated for the subgroup with CMVR and baseline CD4 cell counts <100 cells x 10(6)/l. RESULTS: 1292 patients were started on HAART. 8% of patients had CD4 counts <50 cells x 10(6)/l and 40% had detectable HIV viral load at last follow up. The prevalence of CMVR for the subgroup with baseline CD4 <100 cells x 10(6)/l was 10%. For those with baseline CD4 <100 cells x 10(6)/l, the mean CMVR AE rate was greatest during the first 6 months of follow up after HAART commencement (p <0.003). The mean AE rate per 100 person days at risk was 0.36 (95% CI 0.167 to 0.551) before starting HAART, and 0.14 (95% CI 0.085 to 0.199) after starting HAART (p = 0.03). CONCLUSIONS: HAART significantly prolongs the disease-free intervals in patients with pre-existing disease but recurrences persist within the first 6 months of starting therapy. AE were absent beyond 18 months of follow up in all patients including those with persistently low CD4 counts and detectable HIV viral load indicating clinical immunorestoration. New methods for monitoring the response to therapy are needed to identify those at risk. PMID- 11423460 TI - Bacterial keratitis: a prospective clinical and microbiological study. AB - AIM: To define the clinical and microbiological profile of bacterial keratitis at the Jules Gonin Eye Hospital and to test the in vitro bacterial resistance. METHODS: Patients presenting with bacterial keratitis were prospectively followed; clinical features (age, risk factors, visual acuity) and response to therapy were analysed. Bacteriological profile was determined and the sensitivity/resistance of isolated strains were tested towards 12 ocular antibiotics (NCCLS disc diffusion test). RESULTS: 85 consecutive patients (mean age 44.3 (SD 20.7) years) were prospectively enrolled from 1 March 1997 to 30 November 1998. The following risk factors were identified: contact lens wear, 36%; blepharitis, 21%; trauma, 20%; xerophthalmia, 15%; keratopathies, 8%; and eyelid abnormalities, 6%. The most commonly isolated bacteria were Staphylococcus epidermidis, 40%; Staphylococcus aureus, 22%; Streptococcus pneumoniae, 8%; others Streptococcus species, 5%; Pseudomonas, 9%; Moraxella and Serratia marcescens, 5% each; Bacillus, Corynebacterium, Alcaligenes xyloxidans, Morganella morganii, and Haemophilus influenza, 1% each. 1-15% of strains were resistant to fluoroquinolones, 13-22% to aminoglycosides, 37% to cefazolin, 18% to chloramphenicol, 54% to polymyxin B, 51% to fusidic acid, and 45% to bacitracin. Five of the 85 patients (5.8%) had a poor clinical outcome with a visual loss of one or more lines of visual acuity. CONCLUSION: Fluoroquinolones appear to be the therapy of choice for bacterial keratitis, but, based upon these in vitro studies, some strains may be resistant. PMID- 11423461 TI - Primary iris melanoma: diagnostic features and outcome of conservative surgical treatment. AB - AIMS: To describe features influencing the management of primary iris melanoma and report the outcome of conservative surgical treatment of patients diagnosed with this condition in a tertiary referral academic setting over a 20 year period. METHODS: Retrospective non-comparative case series of consecutive patients diagnosed with iris melanoma from 1980-2000 using medical records from the University of Sydney Department of Ophthalmology and NSW Cancer Registry RESULTS: 51 cases were identified. The most common presentation was growth of a previously noted pigmented lesion. Initial management was either observation or local resection (two had enucleations) with iris reconstruction where possible (23.8%). The mean follow up was 8.7 years (range 1-17 years). Vision of 6/12 or better was maintained in the majority (78.6%) treated by local resection. Pupil reconstruction significantly reduced reported postoperative glare symptoms. Four patients had features suggestive of local recurrence and there was no documented metastatic disease or death from iris melanoma in this series. Histologically, the majority were spindle B cell melanomas. Clinical features including prominent tumour vascularity, rapid growth, and heterogeneous pigmentation were each significantly associated with an epithelioid cell component. Involvement of the iridocorneal angle was frequently associated with ciliary body invasion. CONCLUSIONS: Management decisions for iris melanoma will depend on the clinical features. Mixed or epithelioid histology is more likely in the presence of two or more of the features of malignancy and may justify earlier intervention. When treatment is undertaken, local resection achieves long term tumour clearance with an acceptable morbidity. In resecting iris melanoma, careful assessment for iridocorneal angle involvement is important in treatment planning. Iris reconstruction has a useful role in reducing postoperative photophobia. PMID- 11423462 TI - Academic achievement, close up work parameters, and myopia in Singapore military conscripts. AB - AIM: To determine the relation of refractive error to environmental factors, including close up work, in Singapore military conscripts. METHODS: A cross sectional study was conducted on 429 Singapore military conscripts. Non cycloplegic refraction and A-scan biometry were performed in both eyes. A detailed questionnaire was administered by in-person interview to obtain information about current and past near work activity, extra tuition lessons, educational experiences, and family demographics. RESULTS: Myopia associated with the conscript having been educated in the (gifted, special, or express) educational streams (adjusted odds ratio (OR) = 3.8, 95% confidence interval CI 2.0-7.3), and having completed pre-university education (OR=4.1, 95% CI 1.9-8.8). The reported close up work activity at age 7 years did correlate with age of onset of myopia (p<0.001). In parallel, supplemental tuition lessons in primary school has (OR=2.6, 95% CI 1.4-4.9) associated with conscript myopia. Parental myopia was positively associated with myopia (p<0.001), but this relation disappeared when adjusted for environmental factors. Current (p=0.83) and past close up work activity at age 7 years (p=0.13) did not correlate with myopia. CONCLUSION: Educational level and educational stream positively related to myopia. A relation was observed with reported close up work activity in early childhood and with tuition classes during elementary school, but not with current close up work activity. These results underscore the strong influence of environment in myopia pathogenesis but a role for close up work activity remains indeterminate. PMID- 11423463 TI - Ex vivo adenovirus mediated gene transfection of human conjunctival epithelium. AB - AIM: To investigate the efficacy of "ex vivo" adenoviral vector mediated gene transfection of human conjunctival epithelial cell as a possible route for gene therapy for the distribution of anti-inflammatory agents for the potential treatment of immune mediated ocular inflammatory disorders. METHODS: Human conjunctival cells (HCs) were cultured with various concentrations of recombinant adenoviral vectors carrying a reporter gene LacZ, GFP, or an immunomodulating cytokine vIL-10. vIL-10 in culture supernatant was detected by sandwich ELISA and biological activity was assessed by suppression of ConA stimulated splenocyte proliferation. X-gal and GFP expression was assessed by histochemistry. RESULTS: The extent of adenoviral vector mediated transfer of both reporter genes and vIL 10 was dose dependent. LacZ expression could be detected for at least 50 day after infection with multiple of infection (MOI) 200. Following AdCMVvIL-10 transduction, vIL-10 protein expression occurred between 4-6 days post transduction, and was maintained at a detectable level for at least 1 month. Secreted vIL-10 showed biological activity, significantly inhibiting Con A induced splenocyte proliferation. Additionally, transfection of HCs with two Adv vectors, one carrying LacZ and the other carrying GFP, resulted in co-expression within a single cell. CONCLUSION: These results confirm previous successful adenoviral vector mediated gene transfer to HCs and further show that expression can be maintained. Furthermore the data show HCs can secrete biologically active vIL-10 that could be developed as a strategy to suppress immune mediated disorders. The successful co-transduction of HCs as described for other tissues, opens avenues to develop a multiple target gene therapy locally. PMID- 11423464 TI - Intraocular pressure measurement at the choroid surface: a feasibility study with implications for implantable microsystems. AB - AIMS: To demonstrate that a sensor, which is inserted through the sclera and placed in intimate contact with the choroid, can reliably detect changes in the intraocular pressure (IOP). METHODS: A manometer was used to control the IOP of three cadaver eyes in steps of 7 mm Hg. A piezoresistive pressure sensor was used to measure the pressure at the choroid through a 2.5 mm diameter hole that was surgically removed from the sclera. Data were collected for two configurations; with the sensor: (i) rigidly attached to a miniature positioning stage, and (ii) sutured to the sclera. RESULTS: Both configurations accurately tracked the manometer pressure from 10 mm Hg to 47 mm Hg. For the fixed sensor cases, the average difference between the pressure measured at the choroid and in the anterior chamber was 0.8 mm Hg for the three eyes. For the sutured sensor case, the average difference was 2.1 mm Hg-although a significant portion of this was attributed to an initial offset. The standard deviations at each pressure level for all of the choroid measurements were under 1.0 mm Hg. CONCLUSIONS: Small changes in IOP can be accurately measured by a sensor in contact with the surface of the choroid, for both a fixed sensor configuration and for a sensor sutured to the sclera. These results are the first step in the realisation of a surgically implantable microsensor to monitor IOP for patients suffering from low tension and other difficult to manage forms of glaucoma. PMID- 11423465 TI - Induction of interleukin-8 in human retinal pigment epithelial cells after denuding injury. AB - AIM: To determine interleukin 8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1) expression in response to mechanical injury in human retinal pigment epithelial (HRPE) cells. METHODS: Enzyme linked immunosorbent assay (ELISA) was performed to determine IL-8 and MCP-1 secretion by HRPE cells after mechanical denudation. IL-8 and MCP-1 mRNA expression by HRPE cells was assessed using semiquantitative RT-PCR. The effects of immunosuppressive drugs, dexamethasone (DEX) and cyclosporin A (CSA), as well as immunosuppressive cytokines, interleukin 4 (IL-4), interleukin 10 (IL-10), and interleukin 13 (IL-13), on chemokine expression in HRPE cells after denuding injury were analysed. RESULTS: Mechanical injury induced HRPE IL-8 mRNA and IL-8 secretion. Although MCP-1 mRNA was enhanced slightly after denuding injury, MCP-1 secretion was not increased. DEX and CSA inhibited HRPE chemokine expression after injury. IL-4 and IL-13 enhanced IL-8 and MCP-1 production by HRPE cells after injury while IL-10 had no effect. CONCLUSIONS: These results suggest that IL-8 may be involved in retinal inflammatory responses to injury and that DEX and/or CSA treatment may help control the inflammatory components of retinal diseases such as proliferative vitreoretinopathy. PMID- 11423466 TI - Presence of oestrogen receptor type beta in human retina. AB - BACKGROUND/AIMS: Recent studies have demonstrated the existence of two oestrogen receptor subtypes alpha (ORalpha) and beta (ORbeta) with significant differences of expression among organs. Since important pathologies of human eye could be linked to hormonal status, the expression of ORbeta in ocular posterior segment was sought. METHODS: Immunohistochemical localisation of ORbeta and ORalpha protein and detection of OR mRNAs by reverse transcription-polymerase chain reaction (RT-PCR) were performed in macular and extramacular regions of the retina and in the choroid on male and female donors eyes. RESULTS: ORbeta protein was localised in the ganglion cell layer and in the choroid. At the transcriptional level, mRNA for ORbeta and for ORalpha were both present. Local differences in the expression level were observed, however, suggesting the possibility of variation in the ratio of ORalpha v ORbeta. CONCLUSIONS: The coexistence of two oestrogen receptor subtypes in the human ocular posterior segment raises acute questions about their potential physiological role, but offers a perspective for preferential targeting of a specific receptor subtype. PMID- 11423467 TI - Thrombophilia: genetic polymorphisms and their association with retinal vascular occlusive disease. PMID- 11423468 TI - Raising the suborbicularis fat (SOOF). PMID- 11423469 TI - Impression cytology in the diagnosis of ocular surface squamous neoplasia. PMID- 11423470 TI - Primary iris stromal cyst with pseudohypopyon: an atypical presentation. PMID- 11423471 TI - Hepatocyte nuclear factor-4alpha involved in type 1 maturity-onset diabetes of the young is a novel target of AMP-activated protein kinase. AB - Mutations in the HNF4alpha gene are responsible for type 1 maturity-onset diabetes of the young (MODY1), which is characterized by a defect in insulin secretion. Hepatocyte nuclear factor (HNF)-4alpha is a transcription factor that plays a critical role in the transcriptional regulation of genes involved in glucose metabolism in both hepatocytes and pancreatic beta-cells. Recent evidence has implicated AMP-activated protein kinase (AMPK) in the modulation of both insulin secretion by pancreatic beta-cells and the control of glucose-dependent gene expression in both hepatocytes and beta-cells. Therefore, the question could be raised as to whether AMPK plays a role in these processes by modulating HNF 4alpha function. In this study, we show that activation of AMPK by 5-amino-4 imidazolecarboxamide riboside (AICAR) in hepatocytes greatly diminished HNF 4alpha protein levels and consequently downregulates the expression of HNF-4alpha target genes. Quantitative evaluation of HNF-4alpha target gene expression revealed diminished mRNA levels for HNF-1alpha, GLUT2, L-type pyruvate kinase, aldolase B, apolipoprotein (apo)-B, and apoCIII. Our data clearly demonstrate that the MODY1/HNF-4alpha transcription factor is a novel target of AMPK in hepatocytes. Accordingly, it can be suggested that in pancreatic beta-cells, AMPK also acts by decreasing HNF-4alpha protein level, and therefore insulin secretion. Hence, the possible role of AMPK in the physiopathology of type 2 diabetes should be considered. PMID- 11423472 TI - Transcriptional regulation of plasminogen activator inhibitor type 1 gene by insulin: insights into the signaling pathway. AB - Impairment of the fibrinolytic system, caused primarily by increases in the plasma levels of plasminogen activator inhibitor (PAI) type 1, are frequently found in diabetes and the insulin-resistance syndrome. Among the factors responsible for the increases of PAI-1, insulin has recently attracted attention. In this study, we analyzed the effects of insulin on PAI-1 biosynthesis in HepG2 cells, paying particular attention to the signaling network evoked by this hormone. Experiments performed in CHO cells overexpressing the insulin receptor indicate that insulin increases PAI-1 gene transcription through interaction with its receptor. By using inhibitors of the different signaling pathways evoked by insulin-receptor binding, it has been shown that the biosynthesis of PAI-1 is due to phosphatidylinositol (PI) 3-kinase activation, followed by protein kinase C and ultimately by mitogen-activated protein (MAP) kinase activation and extracellular signal-regulated kinase 2 phosphorylation. We also showed that this pathway is Ras-independent. Transfection of HepG2 cells with several truncations of the PAI-1 promoter coupled to a CAT gene allowed us to recognize two major response elements located in the regions between -804 and -708 and between -211 and -54. Electrophoretic mobility shift assay identified three binding sites for insulin-induced factors, all colocalized with putative Sp1 binding sites. Using supershifting antibodies, the binding of Sp1 could only be confirmed at the binding site located just upstream from the transcription start site of the PAI-1 promoter. A construct comprising four tandem repeat copies of the -93/-62 region of the PAI-1 promoter linked to CAT was transcriptionally activated in HepG2 cells by insulin. These results outline the central role of MAP kinase activation in the regulation of PAI-1 induced by insulin. PMID- 11423473 TI - Novel arguments in favor of the substrate-transport model of glucose-6 phosphatase. AB - The purpose of this work was to discriminate between two models for glucose-6 phosphatase: one in which the enzyme has its catalytic site oriented toward the lumen of the endoplasmic reticulum, requiring transporters for glucose-6 phosphate, inorganic phosphate (Pi), and glucose (substrate-transport model), and a second one in which the hydrolysis of glucose-6-phosphate occurs inside the membrane (conformational model). We show that microsomes preloaded with yeast phosphoglucose isomerase catalyzed the detritiation of [2-(3)H]glucose-6 phosphate and that this reaction was inhibited by up to 90% by S3483, a compound known to inhibit glucose-6-phosphate hydrolysis in intact but not in detergent treated microsomes. These results indicate that glucose-6-phosphate is transported to the lumen of the microsomes in an S3483-sensitive manner. Detritiation by intramicrosomal phosphoglucose isomerase was stimulated twofold by 1 mmol/l vanadate, a phosphatase inhibitor, indicating that glucose-6 phosphatase and the isomerase compete for the same intravesicular pool of glucose 6-phosphate. To investigate the site of release of Pi from glucose-6-phosphate, we incubated microsomes with Pb(2+), which forms an insoluble complex with Pi, preventing its rapid exit from the microsomes. Under these conditions, approximately 80% of the Pi that was formed after 5 min was intramicrosomal, compared with <10% in the absence of Pb(2+). We also show that, when incubated with glucose-6-phosphate and mannitol, glucose-6-phosphatase formed mannitol-1 phosphate and that this nonphysiological product was initially present within the microsomes before being released to the medium. These results indicate that the primary site of product release by glucose-6-phosphatase is the lumen of the endoplasmic reticulum. PMID- 11423474 TI - Liver-derived IGF-I is of importance for normal carbohydrate and lipid metabolism. AB - IGF-I is important for postnatal body growth and exhibits insulin-like effects on carbohydrate metabolism. The function of liver-derived IGF-I is still not established, although we previously demonstrated that liver-derived IGF-I is not required for postnatal body growth. Mice whose IGF-I gene in the liver was inactivated at 24 days of age were used to investigate the long-term role of liver-derived IGF-I for carbohydrate and lipid metabolism. Serum levels of leptin in these mice were increased by >100% at 3 months of age, whereas the fat mass of the mice was decreased by 25% at 13 months of age. The mice became markedly hyperinsulinemic and yet normoglycemic, indicating an adequately compensated insulin resistance. Furthermore, they had increased serum levels of cholesterol. We conclude that liver-derived IGF-I is of importance for carbohydrate and lipid metabolism. PMID- 11423475 TI - Indirect allorecognition in acquired thymic tolerance: induction of donor specific permanent acceptance of rat islets by adoptive transfer of allopeptide pulsed host myeloid and thymic dendritic cells. AB - Pancreatic islet transplantation remains a promising approach to the treatment of type 1 diabetes. Unfortunately, graft failure continues to occur because of immunologic rejection, despite the use of potent immunosuppressive agents. It is therefore reasoned that induction of peripheral tolerance by the use of self dendritic cells (DCs) as a vehicle to deliver specific target antigens to self-T cells without ex vivo manipulation of the recipient is an attractive strategy in the treatment of type 1 diabetes. The finding that intrathymic inoculation of an immunodominant WF major histocompatibility complex (MHC) Class I (RT1.A(u)) peptide five (P5) or P5-pulsed host myeloid DCs induces acquired thymic tolerance raises the possibility that adoptive transfer of allopeptide-primed host myeloid or lymphoid DCs might induce transplant tolerance. To address this hypothesis, we studied the effects of intravenous transfer of in vitro P5-pulsed syngeneic myeloid DCs or in vivo P5-primed syngeneic lymphoid (thymic) DCs on islet survival in the WF-to-ACI rat combination. In vivo primed thymic DCs isolated from ACI rats given intrathymic inoculation of P5 for 2 days were capable of in vitro restimulation of in vivo P5-primed T-cells (memory cells). In the first series of studies, we showed that intravenous-like intrathymic-inoculation of in vitro P5-pulsed host myeloid DCs induced donor-specific permanent acceptance of islets in recipients transiently immunosuppressed with antilymphocyte serum (ALS). We next examined whether thymic DCs isolated from animals that had been previously intrathymically inoculated with P5 could induce T-cell tolerance. The results showed that intravenous adoptive transfer of in vivo P5-primed thymic DCs led to donor-specific permanent acceptance of islets in recipients transiently immunosuppressed with ALS. This finding suggested that the thymic DCs take up and present P5 to developing T-cells to induce T-cell tolerance, thus providing evidence of a direct link between indirect allorecognition and acquired thymic tolerance. The second series of studies examined the mechanisms involved in this model by exploring whether in vivo generation of peptide-specific alloreactive peripheral T-cells by intravenous inoculation of P5-pulsed self-DCs was responsible for the induction of T-cell tolerance. Intrathymic inoculation of splenic T-cells obtained from syngeneic ACI rats primed with intravenous injection of P5-pulsed DCs with a high in vitro proliferative response to P5 in the context of self-MHC induced donor-specific permanent acceptance of islets from WF donors. In addition, the clinically relevant model of intravenous injection of P5-activated T-cells combined with transient ALS immunosuppression similarly induced transplant tolerance, which was then abrogated by thymectomy of the recipient before intravenous injection of the activated T-cells. These data raise the possibility that circulation of peptide-activated T-cells to the host thymus plays a role in the induction and possibly the maintenance of T-cell tolerance in this model. Our findings suggest that intravenous administration of genetically engineered host DCs expressing alloMHC peptides might have therapeutic potential in clinical islet transplantation for the treatment of autoimmune diabetes. PMID- 11423476 TI - Improved glucose tolerance and acinar dysmorphogenesis by targeted expression of transcription factor PDX-1 to the exocrine pancreas. AB - The homeodomain protein PDX-1 is critical for pancreas development and is a key regulator of insulin gene expression. PDX-1 nullizygosity and haploinsufficiency in mice and humans results in pancreatic agenesis and diabetes, respectively. At embryonic day (e) 10.5, PDX-1 is expressed in all pluripotential gut-derived epithelial cells destined to differentiate into the exocrine and endocrine pancreas. At e15, PDX-1 expression is downregulated in exocrine cells, but remains high in endocrine cells. The aim of this study was to determine whether targeted overexpression of PDX-1 to the exocrine compartment of the developing pancreas at e15 would allow for respecification of the exocrine cells. Transgenic (TG) mice were generated in which PDX-1 was expressed in the exocrine pancreas using the exocrine-specific elastase-1 promoter. These mice exhibited a marked dysmorphogenesis of the exocrine pancreas, manifested by increased rates of replication and apoptosis in acinar cells and a progressive fatty infiltration of the exocrine pancreas with age. Interestingly, the TG mice exhibited improved glucose tolerance, but absolute beta-cell mass was not increased. These findings indicate that downregulation of PDX-1 is required for the proper maintenance of the exocrine cell phenotype and that upregulation of PDX-1 in acinar cells affects beta-cell function. The mechanisms underlying these observations remain to be elucidated. PMID- 11423477 TI - Glucagon-like peptide-1 and exendin-4 stimulate beta-cell neogenesis in streptozotocin-treated newborn rats resulting in persistently improved glucose homeostasis at adult age. AB - In neonatal Wistar rats injected with streptozotocin (STZ) at birth (n0-STZ model), a recognized model of beta-cell regeneration, we investigated the capacity of early treatment with glucagon-like peptide 1 (GLP-1) or exendin-4 to promote beta-cell regeneration and thereby improve islet function in the long term, when animals become adults. To this end, n0-STZ rats were submitted to GLP 1 or exendin-4 from postnatal day 2 to day 6 only, and their beta-cell mass and pancreatic functions were tested on day 7 and at 2 months. On day 7, both treatments increased body weight, decreased basal plasma glucose, decreased insulinemia, and increased pancreatic insulin content in n0-STZ rats. At the same age, the beta-cell mass, measured by immunocytochemistry and morphometry methods, was strongly increased in n0-STZ/GLP-1 and n0-STZ/Ex rats compared with n0-STZ rats, representing 51 and 71%, respectively, of the beta-cell mass in Wistar rats, whereas n0-STZ beta-cell mass represented only 21% of the Wistar control value. Despite such early improved beta-cell mass, which is maintained at adult age, the basal and glucose-stimulated insulin secretion (in vivo after intravenous glucose load or in vitro using perfused pancreas) were not improved in the 2-month-old n0-STZ rats previously treated with GLP-1 or exendin-4 compared with untreated n0-STZ rats. However, both treated groups significantly exhibited a decreased basal plasma glucose level and an increased plasma glucose clearance rate compared with the 2-month-old untreated n0-STZ group at adult age. These findings in the n0-STZ model indicate for the first time that GLP-1 or exendin-4 applied during the neonatal diabetic period exert both short- and long term beneficial effects on beta-cell mass recovery and glucose homeostasis. However, the increase in beta-cell mass, which is still present in the adult n0 STZ rats previously treated, contrasts with the poor beta-cell responsiveness to glucose. Further studies are needed to understand the dissociation between beta cell regeneration and the lack of improvement in beta-cell function. PMID- 11423478 TI - Epidermal growth factor increases undifferentiated pancreatic embryonic cells in vitro: a balance between proliferation and differentiation. AB - During embryonic life, the development of a proper mass of mature pancreatic tissue is thought to require the proliferation of precursor cells, followed by their differentiation into endocrine or acinar cells. We investigated whether perturbing the proliferation of precursor cells in vitro could modify the final mass of endocrine tissue that develops. For that purpose, we used activators or inhibitors of signals mediated by receptor tyrosine kinases. We demonstrated that when embryonic day 13.5 rat pancreatic epithelium is cultured in the presence of PD98059, an inhibitor of the mitogen-activated protein (MAP) kinase, epithelial cell proliferation is decreased, whereas endocrine cell differentiation is activated. On the other hand, in the presence of epidermal growth factor (EGF), an activator of the MAP kinase pathway, the mass of tissue that develops is increased, whereas the absolute number of endocrine cells that develops is decreased. Under this last condition, a large number of epithelial cells proliferate but remain undifferentiated. In a second step, when EGF is removed from the pool of immature pancreatic epithelial cells, the cells differentiate en masse into insulin-expressing cells. The total number of insulin-expressing cells that develop can thus be increased by first activating the proliferation of immature epithelial cells with growth factors, thus allowing an increase in the pool of precursor cells, and next allowing their differentiation into endocrine cells by removing the growth factor. This strategy suggests a possible tissue engineering approach to expanding beta-cells. PMID- 11423479 TI - Activation of acetyl-CoA carboxylase by a glutamate- and magnesium-sensitive protein phosphatase in the islet beta-cell. AB - Acetyl-CoA carboxylase (ACC) catalyzes the formation of malonyl-CoA, a precursor in the biosynthesis of long-chain fatty acids, which have been implicated in physiological insulin secretion. The catalytic function of ACC is regulated by phosphorylation (inactive)-dephosphorylation (active). In this study we investigated whether similar regulatory mechanisms exist for ACC in the pancreatic islet beta-cell. ACC was quantitated in normal rat islets, human islets, and clonal beta-cells (HIT-15 or INS-1) using a [(14)C]bicarbonate fixation assay. In the beta-cell lysates, ACC was stimulated by magnesium in a concentration-dependent manner. Of all the dicarboxylic acids tested, only glutamate, albeit ineffective by itself, significantly potentiated magnesium activated ACC in a concentration-dependent manner. ACC stimulation by glutamate and magnesium was maximally demonstrable in the cytosolic fraction; it was markedly reduced by okadaic acid (OKA) in concentrations (<50 nmol/l) that inhibited protein phosphatase 2A (PP2A). Furthermore, pretreatment of the cytosolic fraction with anti-PP2A serum attenuated the glutamate- and magnesium mediated activation of ACC, thereby suggesting that ACC may be regulated by an OKA-sensitive PP2A-like enzyme. Streptavidin-agarose chromatography studies have indicated that glutamate- and magnesium-mediated effects on ACC are attributable to activation of ACC's dephosphorylation; this suggests that the stimulatory effects of glutamate and magnesium on ACC might involve activation of an OKA sensitive PP2A-like enzyme that dephosphorylates and activates ACC. In our study, 5-amino-imidazolecarboxamide (AICA) riboside, a stimulator of AMP kinase, significantly inhibited glucose-mediated activation of ACC and insulin secretion from isolated beta-cells. Together, our data provide evidence for a unique regulatory mechanism for the activation of ACC in the pancreatic beta-cell, leading to the generation of physiological signals that may be relevant for physiological insulin secretion. PMID- 11423480 TI - Dipeptidyl peptidase IV inhibition reduces the degradation and clearance of GIP and potentiates its insulinotropic and antihyperglycemic effects in anesthetized pigs. AB - Glucose-dependent insulinotropic peptide (GIP) is known to be degraded by dipeptidyl peptidase IV (DPP IV), forming an inactive metabolite, but the extent of the enzyme's role in regulating the biological activity of GIP in vivo is still largely unknown. In nonfasted anesthetized pigs given an intravenous infusion of GIP, the intact peptide (determined by a novel NH(2)-terminally directed radioimmunoassay) accounts for only 14.5 +/- 2.5% of total immunoreactivity. This is increased (to 40.9 +/- 0.9%, P < 0.0001) by coadministration of valine-pyrrolidide (a specific DPP IV inhibitor) at a dose that completely inhibits plasma DPP IV activity. The plasma t(1/2) of intact GIP is prolonged by the inhibitor (from 3.3 +/- 0.3 to 8.1 +/- 0.6 min; P < 0.001), whereas the t(1/2) for COOH-terminal immunoreactivity is unaffected (13.2 +/- 0.5 and 11.5 +/- 0.8 min, pre- and postinhibitor). Measurement of arteriovenous concentration differences revealed that the liver, kidney, and extremities are the main sites of removal of exogenous intact GIP (organ extractions, 28.0 +/- 2.2, 26.3 +/- 5.7, and 21.8 +/- 3.0%, respectively). These organ extractions are reduced (P < 0.02) but not eliminated (kidney and extremities) by valine pyrrolidide (to 6.5 +/- 4.6, 14.1 +/- 3.1, and 13.9 +/- 2.4%, respectively). Valine-pyrrolidide potentiates the insulinotropic effect of GIP (P < 0.02), resulting in an enhanced glucose disappearance rate (k, from 8.0 +/- 0.5 to 15.5 +/- 2.2%/min; P < 0.01) and a reduction in the glucose excursion after an intravenous glucose load (area under the curve, from 133 +/- 23 to 75 +/- 9 min. mmol/l; P < 0.05). These results suggest that DPP IV plays an important role in GIP metabolism but is not the sole enzyme responsible for its NH(2)-terminal degradation. Nevertheless, DPP IV inhibition increases the proportion of intact peptide sufficiently to enhance its insulinotropic and antihyperglycemic effects. PMID- 11423481 TI - Gamma globulin levels predict type 2 diabetes in the Pima Indian population. AB - It has been proposed that inflammation or infection may contribute to the development of type 2 diabetes. We examined whether serum gamma globulin, a nonspecific measure of the humoral immune system, predicted changes in glucose tolerance in 2,530 members of the Pima Indian population, a group with a marked predisposition to type 2 diabetes. Cross-sectionally, gamma globulin was positively related to age (r = 0.08, P < 0.0005), BMI (r = 0.09; P < 0.0001), and female sex (P < 0.0001). Gamma globulin concentrations were familial, being positively correlated among siblings (r = 0.23; P < 0.0001) and between parents and their children (mother/child: r = 0.17, P < 0.0001; father/child: r = 0.25, P < 0.0001). Gamma globulin concentrations were higher with greater degrees of American Indian heritage (P < 0.004, with adjustment for age, sex, and BMI) and in the presence of a family history of type 2 diabetes (P < 0.04). Higher gamma globulin levels predicted risk of diabetes. In univariate analysis, a 1 SD difference in gamma globulin was associated with a 20% higher incidence of diabetes in those who were normal glucose tolerant at baseline (hazard rate ratio 1.20 [CI 1.11-1.30]; P < 0.0001) and remained as a significant predictor of diabetes, even when controlled for effects of sex, BMI, and 2-h glucose as additional predictors (hazard rate ratio for 1 SD difference in gamma globulin, 1.14 [1.05-1.24]; P = 0.002). Gamma globulin was also associated in univariate analysis with later development of impaired glucose tolerance (IGT) (hazard rate ratio 1.15 [1.07-1.23]; P < 0.0001), but not with the transition from IGT to diabetes (hazard rate ratio 1.04 [0.90-1.20]; P = 0.6). Thus, gamma globulin levels predict increased risk of diabetes in the Pima population. We suggest that immune function or activation may play a role in the development of type 2 diabetes. PMID- 11423482 TI - Evidence for a major role of skeletal muscle lipolysis in the regulation of lipid oxidation during caloric restriction in vivo. AB - A lipolytic process in skeletal muscle has recently been demonstrated. However, the physiological importance of this process is unknown. We investigated the role of skeletal muscle lipolysis for lipid utilization during caloric restriction in eight obese women before and after 11 days of very low-calorie diet (VLCD) (2.2 MJ per day). Subjects were studied with indirect calorimetry and microdialysis of skeletal muscle and adipose tissue in order to analyze substrate utilization and glycerol (lipolysis index) in connection with a two-step euglycemic hyperinsulinemic (12 and 80 mU/m(2). min) clamp. Local blood flow rates in the two tissues were determined with (133)Xe-clearance. Circulating free fatty acids and glycerol decreased to a similar extent during insulin infusion before and during VLCD, and there was a less marked insulin-induced reduction in lipid oxidation during VLCD. Adipose tissue glycerol release was hampered by insulin infusion to the same extent ( approximately 40%) before and during VLCD. Skeletal muscle glycerol release was not influenced by insulin before VLCD. However, during VLCD insulin caused a marked (fivefold) (P < 0.01) increase in skeletal muscle glycerol release. The effect was accompanied by a fourfold stimulation of skeletal muscle blood flow (P < 0.01). We propose that, during short-term caloric restriction, the reduced ability of insulin to inhibit lipids, despite a preserved antilipolytic effect of the hormone in adipose tissue, is caused by an augmented mobilization of fat from skeletal muscle, and that a physiological role of muscle lipolysis provides a local source of fatty acids. PMID- 11423483 TI - Effects of acute changes of plasma free fatty acids on intramyocellular fat content and insulin resistance in healthy subjects. AB - The reason for the 3- to 4-h delay between a rise in plasma free fatty acid (FFA) levels and the development of insulin resistance remains unknown. In the current study, we have tested the hypothesis that the delay may be caused by the need for plasma FFAs to first enter muscle cells and to be re-esterified there before causing insulin resistance. To this end, we have determined intramyocellular triglyceride (IMCL-TG) content with proton nuclear magnetic resonance ((1)H-NMR) spectroscopy in healthy volunteers before and 4 h after lowering of plasma FFAs (with euglycemic-hyperinsulinemic clamping) or after increasing plasma FFAs (with lipid plus heparin infusions). Increasing plasma FFAs (from 516 to 1,207 micromol/l or from 464 to 1,857 micromol/l, respectively) was associated with acute increases in IMCL-TG from 100 to 109 +/- 5% (P < 0.05) or to 133 +/- 11% (P < 0.01), respectively, and with a significant increase in insulin resistance (P < 0.05 after 3.5 h). Lowering of plasma FFAs from 560 to 41 micromol/l was associated with a tendency for IMCL-TG to decrease (from 100 to 95 +/- 3%). Changes in plasma FFAs correlated linearly with IMCL-TG (r = 0.74, P < 0.003). The demonstration that acute changes in plasma FFAs were accompanied by corresponding changes in IMCL-TG and with the development of insulin resistance, taken together with previous reports of a close correlation between IMCL-TG and insulin resistance, supported the notion that accumulation of IMCL-TG is a step in the development of FFA-induced insulin resistance. PMID- 11423484 TI - The effect of acute hypoglycemia on brain function and activation: a functional magnetic resonance imaging study. AB - The authors' aim was to examine the regional anatomy of brain activation by cognitive tasks commonly used in hypoglycemia research and to assess the effect of acute hypoglycemia on these in healthy volunteers. Eight right-handed volunteers performed a set of cognitive tasks-finger tapping (FT), simple reaction time (SRT), and four-choice reaction time (4CRT)-twice during blood oxygen level-dependent (BOLD) functional magnetic resonance imaging of the brain on two occasions. In study 1 (n = 6), plasma glucose was maintained at euglycemia (5 mmol/l) throughout. In study 2 (n = 6), plasma glucose was reduced to 2.5 mmol/l for the second set. Performance of the tasks resulted in specific group brain activation maps. During hypoglycemia, FT slowed (P = 0.026), with decreased BOLD activation in right premotor cortex and supplementary motor area and left hippocampus and with increased BOLD activation in left cerebellum and right frontal pole. Although there was no significant change in SRT, BOLD activation was reduced in right cerebellum and visual cortex. The 4CRT deteriorated (P = 0.020), with reduction in BOLD activation in motor and visual systems but increased BOLD signal in a large area of the left parietal association cortex, a region involved in planning. Hypoglycemia impairs simple brain functions and is associated with task-specific localized reductions in brain activation. For a task with greater cognitive load, the increased BOLD signal in planning areas is compatible with recruitment of brain regions in an attempt to limit dysfunction. Further investigation of these mechanisms may help devise rational treatment strategies to limit cortical dysfunction during acute iatrogenic hypoglycemia. PMID- 11423485 TI - Glucose effects on skin keratinocytes: implications for diabetes skin complications. AB - Altered skin wound healing is a common cause of morbidity and mortality among diabetic patients. However, the molecular mechanisms whereby diabetes alters skin physiology have not been elucidated. In this study, we investigated the relative roles of hyperglycemia, insulin, and IGF-I, all of which are abnormal in diabetes, in primary murine skin keratinocytes. These cells proliferate and differentiate in vitro in a manner similar to skin in vivo. It was found that in the presence of high glucose (20 mmol/l), the glucose transport rate of primary proliferating or differentiating keratinocytes was downregulated, whereas at 2 mmol/l glucose, the transport rate was increased. These changes were associated with changes in the GLUT1 expression and with changes in the affinity constant (K(m)) of the transport. Exposure to high glucose was associated with changes in cellular morphology, as well as with decreased proliferation and enhancement of Ca(2+)-induced differentiation of keratinocytes. Furthermore, in the presence of high glucose, ligand-induced IGF-I receptor but not insulin receptor (IR) autophosphorylation was decreased. Consequently, in high glucose, the effects of IGF-I on glucose uptake and keratinocyte proliferation were inhibited. Interestingly, lack of IR expression in IR-null keratinocytes abolished insulin induced glucose uptake and partially decreased insulin- and IGF-I-induced proliferation, demonstrating the direct involvement of the IR in these processes. Our results demonstrate that hyperglycemia and impaired insulin signaling might be directly involved in the development of chronic complications of diabetes by impairing glucose utilization of skin keratinocytes as well as skin proliferation and differentiation. PMID- 11423486 TI - Pharmacological inhibition of diabetic retinopathy: aminoguanidine and aspirin. AB - Effects of aminoguanidine and aspirin on the development of retinopathy have been examined in 5-year studies of diabetic dogs. Either agent was administered daily in doses of 20-25 mg. kg(-1). day(-1). Because severity of hyperglycemia greatly influences development of the retinopathy, special effort was devoted to maintaining comparable glycemia in experimental and control groups. The retinal vasculature was isolated by the trypsin digest method, and retinopathy was assessed by light microscopy. Diabetes for 5 years resulted, as expected, in saccular capillary aneurysms, pericyte ghosts, acellular capillaries, retinal hemorrhages, and other lesions. Administration of aminoguanidine essentially prevented the retinopathy, significantly inhibiting the development of retinal microaneurysms, acellular capillaries, and pericyte ghosts compared with diabetic controls. Aspirin significantly inhibited the development of retinal hemorrhages and acellular capillaries over the 5 years of study, but had less effect on other lesions. Although diabetes resulted in significantly increased levels of advanced glycation end products (AGEs) (namely, pentosidine in tail collagen and aorta, and Hb-AGE), aminoguanidine had no significant influence on these parameters of glycation. Nitration of a retinal protein was significantly increased in diabetes and inhibited by aminoguanidine. The biochemical mechanism by which aminoguanidine has inhibited retinopathy thus is not clear. Aminoguanidine (but not aspirin) inhibited a diabetes-induced defect in ulnar nerve conduction velocity, but neither agent was found to influence kidney structure or albumen excretion. PMID- 11423487 TI - Lipoprotein lipase mediates the uptake of glycated LDL in fibroblasts, endothelial cells, and macrophages. AB - The nonenzymatic glycation of LDL is a naturally occurring chemical modification of apolipoprotein (apo)-B lysine residues by glucose. Once glycated, LDL is only poorly recognized by lipoprotein receptors including the LDL receptor (LDL-R), the LDL-R-related protein (LRP), and scavenger receptors. Glycated LDL (gLDL) is a preferred target for oxidative modifications. Additionally, its presence initiates different processes that can be considered "proatherogenic." Thus, LDL glycation might contribute to the increased atherosclerotic risk of patients with diabetes and familial hypercholesterolemia. Here we investigate whether lipoprotein lipase (LPL) can mediate the cellular uptake of gLDL. The addition of exogenous LPL to the culture medium of human skin fibroblasts, porcine aortic endothelial cells, and mouse peritoneal macrophages enhanced the binding, uptake, and degradation of gLDL markedly, and the relative effect of LPL on lipoprotein uptake increased with the degree of apoB glycation. The efficient uptake of gLDL by LDL-R-deficient fibroblasts and LRP-deficient Chinese hamster ovary cells in the presence of LPL suggested a mechanism that was independent of the LDL-R and LRP. In macrophages, the uptake of gLDL was also correlated with their ability to produce LPL endogenously. Mouse peritoneal macrophages from genetically modified mice, which lacked LPL, exhibited a 75% reduction of gLDL uptake compared with normal macrophages. The LPL-mediated effect required the association of the enzyme with cell surface glycosaminoglycans but was independent of its enzymatic activity. The uptake of gLDL in different cell types by an LPL-mediated process might have important implications for the cellular response after gLDL exposure as well as the removal of gLDL from the circulation. PMID- 11423488 TI - Diabetes-induced accelerated atherosclerosis in swine. AB - Patients with diabetes are at higher risk for atherosclerotic disease than nondiabetic individuals with other comparable risk factors. Studies examining mechanisms underlying diabetes-accelerated atherosclerosis have been limited by the lack of suitable humanoid animal models. In this study, diabetes was superimposed on a well-characterized swine model of atherosclerosis by injection of the beta-cell cytotoxin streptozotocin (STZ), resulting in a >80% reduction in beta-cells and an increase in plasma glucose to diabetic levels. Animals were maintained without exogenous insulin for up to 48 weeks. Plasma glucose and cholesterol levels and lesion extent and severity were quantified in swine with diabetes and hyperlipemia alone and in combination compared with controls. Diabetes had no effect on plasma cholesterol levels, but diabetic/hyperlipemic (D HL) swine developed hypertriglyceridemia and showed a doubling in aortic sudanophilia over nondiabetic/hyperlipemic (N-HL) swine as early as 12 weeks (47.25 +/- 4.5 vs. 24.0 +/- 4.6%). At 20 weeks, coronary artery stenosis was significantly greater in D-HL than in N-HL animals (86 +/- 10 vs. 46 +/- 8%). Coronary lesions predominantly arose in the first 2-3 cm of the vessels and displayed humanoid morphology. Aortic lesions in D-HL swine had double the cholesterol content of those in N-HL swine, and incorporation of oleate into cholesteryl ester was significantly greater in grossly normal aortic areas of D HL swine compared with N-HL and was attributed to similar elevated incorporation in monocytes. This large study demonstrates that a model of diabetes with humanoid characteristics, including hypertriglyceridemia and severe, accelerated atherosclerosis can be reproducibly induced and maintained in swine. This model should potentially be of great value in elucidating mechanisms underlying the accelerated atherosclerosis seen in human diabetic individuals. PMID- 11423489 TI - Actin and annexins I and II are among the main endothelial plasmalemma-associated proteins forming early glucose adducts in experimental diabetes. AB - An immunochemical and biochemical study was performed to reveal which of the endothelial plasma membrane proteins become glycated during the early phases of diabetes. The blood front of the lung microvascular endothelial plasmalemma was purified by the cationic colloidal silica method from normal and diabetic (streptozotocin-induced) rats and comparatively analyzed by two-dimensional electrophoresis. No major qualitative differences in the general spectrum of endothelial plasmalemmal proteins were recorded between normoglycemic and hyperglycemic animals. By probing with anti-glucitollysine antibodies, we found that at 1 month after the onset of diabetes, several endothelial membrane polypeptides contained glucose covalently linked to their lysyl residues. Ten days of insulin treatment restored euglycemia in the diabetic animals and completely abolished the membrane nonenzymatic glycosylation. All the glycated polypeptides of the endothelial plasma membrane belong to the peripheral type and are associated with its cytoplasmic face (cell cortex). They were solubilized by buffers of high pH and were not detected in the lung cytosolic fraction (100,000 g). By microsequencing, the major proteins labeled by the anti-glucitollysine have been identified as being actin, annexin I, annexin II, the p34 subunit of the Arp2/3 complex, and the Ras suppressor protein-1. Conversely, the intrinsic endothelial membrane proteins do not seem to be affected by hyperglycemia. This defines the internal face of the endothelial plasma membrane, particularly the cortical cytoskeleton, as a preferential target for nonenzymatic glycosylation in diabetes, with possible consequences on the fluidity of the endothelial plasmalemma and impairment of the endothelial mechanotransducing ability. PMID- 11423490 TI - The 75-g oral glucose tolerance test in pregnancy. PMID- 11423491 TI - Declining insulin requirement in the late first trimester of diabetic pregnancy. AB - OBJECTIVE: To investigate whether pregnancies complicated by type 1 diabetes are associated with a decrease in first-trimester insulin requirement. RESEARCH DESIGN AND METHODS: We examined the weekly insulin requirement (as units per kilogram per day) during the first trimester of pregnancy in diabetic women in the Diabetes in Early Pregnancy Study (DIEP) with accurate gestational dating, regular glucose monitoring, daily insulin-dose recording, and monthly glycohemoglobin measurements. RESULTS: In pregnancies that resulted in live-born full-term singleton infants, a significant 18% increase in mean weekly dosage was observed between weeks 3 and 7 (P = 0.000), followed by a significant 9% decline from week 7 through week 15 (P = 0.000). Further testing localized a significant change in insulin dose in the interval beginning weeks 7-8 and ending weeks 11-12 (P = 0.014). Within this interval, the maximum decrease was between weeks 9 and 10 (mean), 10 and 11 (median), and 8 and 9 (most frequent maximal decrease). To determine whether prior poor glucose control exaggerated these trends, we categorized the women based on their glycohemoglobin values: <2 SDs above the mean of a normal population (subgroup 1), 2-4 SDs (subgroup 2), and >4 SDs (subgroup 3) at baseline. Late first-trimester declines in dosage were statistically significant in subgroup 2 (P = 0.002) and subgroups 2 and 3 together (P = 0.003). Similarly, women with BMI >27.0 had a greater initial insulin rise and then fall compared with leaner women. CONCLUSIONS: Observations in the DIEP cohort disclose a mid-first-trimester decline in insulin requirement in type 1 diabetic pregnant women. Possible explanations include overinsulinization of previously poorly controlled diabetes, a transient decline in progesterone secretion during the late first-trimester luteo-placental shift in progesterone secretion, or other hormonal shifts. Clinicians should anticipate a clinically meaningful reduction in insulin requirement in the 5-week interval between weeks 7 and 12 of gestation. PMID- 11423492 TI - The effect of flexible low glycemic index dietary advice versus measured carbohydrate exchange diets on glycemic control in children with type 1 diabetes. AB - OBJECTIVE: To determine the long-term effect of low glycemic index dietary advice on metabolic control and quality of life in children with type 1 diabetes. RESEARCH DESIGN AND METHODS: Children with type 1 diabetes (n = 104) were recruited to a prospective, stratified, randomized, parallel study to examine the effects of a measured carbohydrate exchange (CHOx) diet versus a more flexible low-glycemic index (GI) dietary regimen on HbA(1c) levels, incidence of hypo- and hyperglycemia, insulin dose, dietary intake, and measures of quality of life over 12 months. RESULTS: At 12 months, children in the low-GI group had significantly better HbA(1c) levels than those in the CHOx group (8.05 +/- 0.95 vs. 8.61 +/- 1.37%, P = 0.05). Rates of excessive hyperglycemia (>15 episodes per month) were significantly lower in the low-GI group (35 vs. 66%, P = 0.006). There were no differences in insulin dose, hypoglycemic episodes, or dietary composition. The low-GI dietary regimen was associated with better quality of life for both children and parents. CONCLUSIONS: Flexible dietary instruction based on the food pyramid with an emphasis of low-GI foods improves HbA(1c) levels without increasing the risk of hypoglycemia and enhances the quality of life in children with diabetes. PMID- 11423493 TI - Hormone replacement therapy is associated with better glycemic control in women with type 2 diabetes: The Northern California Kaiser Permanente Diabetes Registry. AB - OBJECTIVE: In women with diabetes, the changes that accompany menopause may further diminish glycemic control. Little is known about how hormone replacement therapy (HRT) affects glucose metabolism in diabetes. The aim of this study was to examine whether HbA(1c) levels varied by current HRT among women with type 2 diabetes. RESEARCH DESIGN AND METHODS: In a cohort of 15,435 women with type 2 diabetes who were members of a health maintenance organization, HbA(1c) and HRT were assessed by reviewing records in the health plan's computerized laboratory and pharmacy systems. Sociodemographic and clinical information were collected by survey. RESULTS: The mean age was 64.7 years (SD +/- 8.7). The study cohort comprised 55% non-Hispanic whites, 14% non-Hispanic blacks, 12% Hispanics, 11% Asians, 4% "other" ethnic groups, and 4% with missing ethnicity data. Current HRT was observed in 25% of women. HbA(1c) levels were significantly lower in women currently using HRT than in women not using HRT (age-adjusted mean +/- SE: 7.9 +/ 0.03 vs. 8.5 +/- 0.02, respectively, P = 0.0001). No differences in HbA(1c) level were observed between women using unopposed estrogens and women using opposed estrogens. In a Generalized Estimating Equation model, which took into account patient clustering within physician and adjusted for age, ethnicity, education, obesity, hypoglycemic therapy, diabetes duration, self-monitoring of blood glucose, and exercise, HRT remained significantly and independently associated with decreased HbA(1c) levels (P = 0.0001). CONCLUSIONS: HRT was independently associated with decreased HbA(1c) level. Clinical trials will be necessary to understand whether HRT may improve glycemic control in women with diabetes. PMID- 11423494 TI - Gestational diabetes mellitus diagnosed with a 2-h 75-g oral glucose tolerance test and adverse pregnancy outcomes. AB - OBJECTIVE: To evaluate American Diabetes Association (ADA) and World Health Organization (WHO) diagnostic criteria for gestational diabetes mellitus (GDM) against pregnancy outcomes. RESEARCH DESIGN AND METHODS: This cohort study consecutively enrolled Brazilian adult women attending general prenatal clinics. All women were requested to undertake a standardized 2-h 75-g oral glucose tolerance test (OGTT) between their estimated 24th and 28th gestational weeks and were then followed to delivery. New ADA criteria for GDM require two plasma glucose values > or = 5.3 mmol/l (fasting), > or = 10 mmol/l (1 h), and > or = 8.6 mmol/l (2 h). WHO criteria require a plasma glucose > or = 7.0 mmol/l (fasting) or > or = 7.8 mmol/l (2 h). Individuals with hyperglycemia indicative of diabetes outside of pregnancy were excluded. RESULTS: Among the 4,977 women studied, 2.4% (95% CI 2.0-2.9) presented with GDM by ADA criteria and 7.2% (6.5 7.9) by WHO criteria. After adjustment for the effects of age, obesity, and other risk factors, GDM by ADA criteria predicted an increased risk of macrosomia (RR 1.29, 95% CI 0.73-2.18), preeclampsia (2.28, 1.22-4.16), and perinatal death (3.10, 1.42-6.47). Similarly, GDM by WHO criteria predicted increased risk for macrosomia (1.45, 1.06-1.95), preeclampsia (1.94, 1.22-3.03), and perinatal death (1.59, 0.86-2.90). Of women positive by WHO criteria, 260 (73%) were negative by ADA criteria. Conversely, 22 (18%) women positive by ADA criteria were negative by WHO criteria. CONCLUSIONS: GDM based on a 2-h 75-g OGTT defined by either WHO or ADA criteria predicts adverse pregnancy outcomes. PMID- 11423495 TI - The relationship of fasting plasma glucose values and other variables to 2-h postload plasma glucose in Japanese subjects. AB - OBJECTIVE: To investigate the relationship between fasting plasma glucose (FPG) values and other variables (e.g., age, sex, and BMI) to 2-h post-75-g oral glucose load glycemia (PG) in Japanese subjects. RESEARCH DESIGN AND METHODS: Subjects included 13,694 Japanese subjects between 20 and 83 years of age (10,677 men and 3,017 women) who were undergoing a 75-g oral glucose tolerance test (OGTT) during a health screening performed at our hospital. The influences of age for 2-h PG at a fixed fasting plasma glucose (FPG) level of 126 mg/dl were analyzed. Multiple linear regression analysis was performed using a model in which the dependent variable was 2-h PG using the following explanatory variables: FPG, age, sex, BMI, blood pressure, plasma cholesterol, and triglyceride (TG) levels. RESULTS: The 2-h PG at a fixed FPG of 126 mg/dl increased by 0.94 mg/dl per year in patients aged between 30 and 78 years (r = 0.68, P < 0.0001). In multiple regression, five explanatory variables (FPG, age, BMI, plasma TG levels, and systolic blood pressure levels) were all positively associated with 2-h PG. The percentages of patients with 2-h diabetes (isolated postchallenge hyperglycemia [IPH]) versus fasting plus 2-h diabetes by the World Health Organization criteria significantly (P = 0.005) increased as the patients' decades increased, whereas the impact of BMI on the percentages was significant only in young patients (P = 0.001). CONCLUSIONS: Aging was found to be the second best predictor of 2-h PG on multiple regression. Therefore, OGTT should be performed especially in elderly patients because they show IPH more frequently. PMID- 11423496 TI - Association of the Ala54-Thr polymorphism in the intestinal fatty acid-binding protein with 2-h postchallenge insulin levels in the Framingham Offspring Study. AB - OBJECTIVE: To investigate the association of variants of the intestinal fatty acid-binding protein gene (FABP2) with fasting and postchallenge glucose and insulin levels, HbA(1c), and prevalence of type 2 diabetes in a separate sample of men and women. RESEARCH DESIGN AND METHODS: Subjects were participants in the Framingham Offspring Study, a long-term community-based prospective observational study of risk factors for cardiovascular disease. The study sample consisted of 762 men and 922 women. RESULTS: In women, carriers of the Thr54 allele had significantly higher 2-h postchallenge insulin levels than noncarriers (104.4 +/- 73.0 vs. 93.4 +/- 61.5 microU/ml; P = 0.0139). This relationship remained significant after adjustment for familial relationship, age, BMI, triglycerides, APOE genotype, smoking, alcohol intake, the use of beta-blockers, menopausal status, and estrogen therapy. No such significant association was observed in men. In both men and women, there were no statistical associations between the FABP2 polymorphism and BMI, fasting glucose, fasting insulin, 2-h postchallenge glucose levels, HbA(1c), and prevalence of type 2 diabetes. CONCLUSIONS: These results suggest that the FABP2 Thr54 allele may have a minor contribution to the insulin resistance syndrome in a white general population. PMID- 11423497 TI - Relationship between ethnicity and glycemic control, lipid profiles, and blood pressure during the first 9 years of type 2 diabetes: U.K. Prospective Diabetes Study (UKPDS 55). AB - OBJECTIVE: To assess the relationship among self-reported ethnicity, metabolic control, and blood pressure during treatment of type 2 diabetes. RESEARCH DESIGN AND METHODS: We studied 2,999 newly diagnosed type 2 diabetic patients recruited to the U.K. Prospective Diabetes Study who were randomized to conventional or intensive glucose control policies if their fasting plasma glucose levels remained >6 mmol/l after a dietary run-in. A total of 2,484 patients (83%) were white Caucasian (WC), 265 patients (9%) were Afro-Caribbean (AC), and 250 patients (8%) were Asian of Indian origin (IA). Variables were assessed at 3, 6, and 9 years. RESULTS: During the 9-year study period, body weight increased more in WC patients (mean 5.0 kg) than in AC (3.0 kg) and IA (2.5 kg) patients (P < 0.001). After adjusting for age, sex, baseline value, treatment allocation, and change in weight, there were no consistent ethnic differences in mean change in fasting plasma glucose or HbA(1c). After adjustment for antihypertensive therapy, increase in systolic blood pressure at 9 years was greatest in AC patients (7 mmHg; P < 0.01 vs. WC patients). Mean diastolic blood pressure, total cholesterol, and LDL cholesterol decreased progressively during the 9 years in each group. In AC patients, the mean increase in HDL cholesterol (0.16 mmol/l) at 3 years, maintained to 9 years, and the mean decrease in plasma triglyceride level (0.4 mmol/l) at 9 years were greater than in WC and IA patients (P < 0.001). CONCLUSIONS: This study shows important ethnic differences in body weight, lipid profiles, and blood pressure, but not glycemic control, during 9 years after diagnosis of type 2 diabetes. AC patients maintained the most favorable lipid profiles, but hypertension developed in more AC patients than WC or IA patients. Ethnicity-specific glycemic control of type 2 diabetes seems unnecessary, but other risk factors need to be addressed independently. PMID- 11423498 TI - Association of physical activity and serum insulin concentrations in two populations at high risk for type 2 diabetes but differing by BMI. AB - OBJECTIVE: Physical activity and insulin sensitivity are related in epidemiological studies, but the consistency of this finding among populations that greatly differ in body size is uncertain. The present multiethnic epidemiological study examined whether physical activity was related to insulin concentrations in two populations at high risk for diabetes that greatly differ by location, ethnic group, and BMI. RESEARCH DESIGN AND METHODS: The study populations consisted of 2,321 nondiabetic Pima Indian men and women aged 15-59 years from Arizona and 2,716 nondiabetic men and women aged 35-54 years from Mauritius. Insulin sensitivity was estimated by mean insulin concentration (average of the fasting and postload insulin), and total (i.e., leisure and occupational) physical activity was assessed by questionnaire. RESULTS: Pima men and women who were more active had significantly (P < 0.05) lower mean insulin concentrations than those less active (BMI and age-adjusted means were 179 vs. 200 and 237 vs. 268 pmol/l). Similar findings were noted in Mauritian men and women (94 vs. 122 and 127 vs. 148 pmol/l). In both populations, activity remained significantly associated with mean insulin concentration controlled for age, BMI, waist-to-thigh or waist-to-hip ratio, and mean glucose concentrations. CONCLUSIONS: Physical activity was negatively associated with insulin concentrations both in the Pima Indians, who tend to be overweight, and in Mauritians, who are leaner. These findings suggest a beneficial role of activity on insulin sensitivity that is separate from any influence of activity on body composition. PMID- 11423499 TI - Autoantibodies to a 38-kDa glycosylated islet cell membrane-associated antigen in (pre)type 1 diabetes: association with IA-2 and islet cell autoantibodies. AB - OBJECTIVE: To study the association of autoantibodies against a 38-kDa glycated islet cell membrane-associated (GLIMA) protein with (pre)type 1 diabetes, patient characteristics, and other immune and genetic markers of the disease and to evaluate the possible added value of GLIMA antibody determinations for disease prediction and classification. RESEARCH DESIGN AND METHODS: Recent-onset type 1 diabetic patients (n = 100), prediabetic siblings (n = 23), and nondiabetic control subjects (n = 100) were consecutively recruited by the Belgian Diabetes Registry. GLIMA antibodies were determined by immunoprecipitation of radiolabeled islet cell proteins; islet cell antibodies (ICAs) were determined by indirect immunofluorescence; and insulin autoantibodies (IAAs), insulinoma-associated protein-2 antibodies (IA-2As), and GAD antibodies (GADAs) were determined by radioligand assays. RESULTS: GLIMA antibodies were detected in 38% of type 1 diabetic patients and 35% of prediabetic siblings (during follow-up) vs. 0% in control subjects (P < 0.001). Their prevalence was lower than that of other antibodies and was significantly associated with high levels of IA-2A and ICA (P < 0.0001). In (pre)diabetes, GLIMA antibodies could only be demonstrated in sera positive for > or = 1 other autoantibody. CONCLUSIONS: GLIMA antibodies are strongly associated with type 1 diabetes and antibody markers of rapid progression to clinical onset but have a lower diagnostic sensitivity for the disease than IAA, ICA, IA-2A, or GADA. In its present form, the GLIMA antibody assay does not provide much additional information for prediction or classification of diabetes, compared with that obtained from the measurement of IA-2As alone or in combination with IAAs, ICAs, and GADAs. PMID- 11423500 TI - Role of hemochromatosis C282Y and H63D mutations in HFE gene in development of type 2 diabetes and diabetic nephropathy. AB - OBJECTIVE: In patients with clinical hemochromatosis, the frequency of diabetes ranges from 20 to 50%, and the heterozygosity for the C282Y mutation in the HFE gene might be associated with an increased risk for diabetes. There are also some reports that suggest that iron overload might cause diabetic nephropathy. RESEARCH DESIGN AND METHODS: We performed an association study to assess the role of the C282Y and H63D mutations in the HFE gene as a risk factor for type 2 diabetes and diabetic nephropathy. Altogether, 563 patients with type 2 diabetes were included in the study. In the analyzed group, 108 patients had overt proteinuria, 154 had microalbuminuria, and 301 had normoalbuminuria. Among the patients with normoalbuminuria, only those with known diabetes duration > or = 10 years were considered normoalbuminuric (n = 162). A total of 196 unrelated healthy subjects were used as a control group. All subjects were genotyped for C282Y and H63D using the polymerase chain reaction-based protocol. RESULTS: There was an increased frequency of 282Y allele carriers among patients with type 2 diabetes versus healthy control subjects (OR 5.3, 95% CI 1.6-17.3). We observed an increased frequency of the 63D allele carriers among patients with diabetic nephropathy (1.8, 1.2-2.8). CONCLUSIONS: In conclusion, our study is the first to indicate that being a carrier of the H63D hemochromatosis mutation is a risk factor for nephropathy in type 2 diabetic patients. We also confirmed previous observations that the frequency of the 282Y mutation was higher in patients with type 2 diabetes than it was in the general population of healthy subjects. PMID- 11423501 TI - Type 1 and type 2 diabetes and incident hip fractures in postmenopausal women. AB - OBJECTIVE: To examine whether postmenopausal women with diabetes experienced a higher incidence of hip fracture than women without diabetes. RESEARCH DESIGN AND METHODS: A prospective cohort of 32,089 postmenopausal women residing in Iowa were surveyed by mail in 1986 and followed for 11 years. Diabetes status and other potential risk factors were assessed by questionnaires at baseline; incidence of hip fracture was ascertained by follow-up questionnaires. RESULTS: A total of 490 hip fractures were reported over 306,900 person-years of follow-up. After adjustment for age, smoking status, estrogen use, BMI, and waist-to-hip ratio, women with type 1 diabetes (n = 47) were 12.25 times (95% CI 5.05-29.73) more likely to report an incident hip fracture than women without diabetes. Women with type 2 diabetes had a 1.70-fold higher risk (1.21-2.38) of incident hip fracture than women without diabetes. Longer duration of type 2 diabetes was associated with higher incidence, as was use of insulin or oral diabetes medications in women with type 2 diabetes. Furthermore, women who were initially free of diabetes but in whom diabetes developed had a relative risk of hip fracture of 1.60 (1.14-2.25) compared with women who never had diabetes. CONCLUSIONS: Postmenopausal women who have diabetes or in whom diabetes develops are at higher risk for hip fracture than nondiabetic postmenopausal women. Strategies to prevent osteoporosis and/or falling may be especially warranted in women with diabetes. PMID- 11423502 TI - Diabetes and risk of fracture: The Blue Mountains Eye Study. AB - OBJECTIVE: To examine associations between measures of diabetes and risk of fracture in a population-based sample of older Australians. RESEARCH DESIGN AND METHODS: This was a prospective study of 3,654 subjects aged 49 years and older who were residents in the Blue Mountains, west of Sydney, Australia. At baseline, subjects were asked questions about history and treatment of diabetes, and fasting blood samples were taken. Photographs were taken of the retina and lens to grade retinopathy and cataract. Details of fractures (excluding rib and vertebral fractures) were collected by a combination of self-report and medical record searches; all fractures were radiologically confirmed. RESULTS: After 2 years of follow-up, we found that several diabetes-related factors were significantly associated (in multivariate models) with increased risk of all fractures combined, including presence of diabetic retinopathy (adjusted RR 5.4, 95% CI 2.7-10.8), diabetes duration > or = 10 years (3.3, 1.3-8.2), cortical cataract involving > or = 25% of the lens area (2.5, 1.3-4.7), and insulin treatment (5.9, 2.6-13.5). The proximal humerus was the only individual fracture site associated with diabetes. Diabetic retinopathy (10.3, 2.2-48.0), diabetes duration (for > or = 10 years duration; 11.4, 2.4-54.2), and insulin treatment (18.8, 4.0-88.7) were all associated with proximal humerus fracture. CONCLUSIONS: These data suggest a significantly increased risk of fracture associated with diabetic retinopathy, advanced cortical cataract, longer diabetes duration, and insulin treatment. However, there are some shortcomings in this study that may limit these findings. PMID- 11423503 TI - Diabetes and diabetic retinopathy in a Mexican-American population: Proyecto VER. AB - OBJECTIVES: The prevalence rate of diabetes is probably higher in Hispanics than in Caucasians, although there is controversy about differences in the risk of diabetic retinopathy. The purpose of the study is to determine the prevalence rates of diabetes and diabetic retinopathy in a population-based study of Hispanics aged > or = 40 years. RESEARCH DESIGN AND METHODS: Proyecto VER is a random sample of Hispanic populations aged > or = 40 years in Arizona. A total of 4,774 individuals (71.6% of the eligible sample) completed the examinations. Diabetes was defined as self-report of a physician diagnosis or HbA(1c) value of > or = 7.0%. Diabetic retinopathy was assessed on stereo fundus photographs of fields 1, 2, and 4. RESULTS: The prevalence rate of diabetes in the Hispanic community (individuals > or = 40 years of age) was 22%. The prevalence rate of diabetic retinopathy (DR) was 48%; 32% had moderate to severe nonproliferative and proliferative retinopathy. DR increased with increasing duration of diabetes and increasing level of HbA(1c). The prevalence rate of DR-like changes in the sample of individuals without diabetic retinopathy was 15% and was not associated with hypertension, systolic blood pressure, or diastolic blood pressure. CONCLUSIONS: The prevalence rate of diabetes in this population of Hispanics is high, almost twice that of Caucasians. The prevalence rate of DR is high but similar to reports in a Caucasian population. The prevalence rate of 9% moderate to severe retinopathy in the newly diagnosed group suggests that efforts to improve detection and treatment of diabetes in Hispanics may be warranted. PMID- 11423504 TI - Risk of type 1 diabetes development in children with incidental hyperglycemia: A multicenter Italian study. AB - OBJECTIVE: The aim of our study was to determine whether children with incidental hyperglycemia are at an increased risk of developing type 1 diabetes. RESEARCH DESIGN AND METHODS: A total of 748 subjects, 1-18 years of age (9.04 +/- 3.62, mean +/- SD), without family history of type 1 diabetes, without obesity, and not receiving drugs were studied and found to have incidental elevated glycemia defined as fasting plasma glucose >5.6 mmol/l confirmed on two occasions. Subjects were tested for immunological, metabolic, and immunogenetic markers. RESULTS: Islet cell antibodies >5 Juvenile Diabetes Foundation units were found in 10% of subjects, elevated insulin autoantibody levels in 4.6%, GAD antibody in 4.9%, and anti-tyrosine phosphatase-like protein autoantibodies in 3.9%. First phase insulin response (FPIR) was <1st centile in 25.6% of subjects. The HLA DR3/DR3 and HLA-DR4/other alleles were more frequent in hyperglycemic children than in normal control subjects (P = 0.012 and P = 0.005, respectively), and the HLA-DR other/other allele was less frequent than in normal control subjects (P = 0.000027). After a median follow-up of 42 months (range 1 month to 7 years), 16 (2.1%) subjects (11 males and 5 females), 4.1-13.9 years of age, became insulin dependent. All had one or more islet autoantibodies, and the majority had impaired insulin response and genetic susceptibility to type 1 diabetes. Diabetes symptoms were recorded in 11 patients and ketonuria only in 4 patients. The cumulative risk of type 1 diabetes was similar in males and females, and it was also similar in subjects under or over 10 years, whereas the cumulative risk of type 1 diabetes was increased in subjects with one or more autoantibodies and in those with FPIR <1st centile. CONCLUSIONS: Children with incidental hyperglycemia have a higher-than-normal frequency of immunological, metabolic, or genetic markers for type 1 diabetes and have an increased risk of developing type 1 diabetes. PMID- 11423505 TI - Use of an automated device for alternative site blood glucose monitoring. AB - OBJECTIVE: To evaluate the accuracy, comfort, and ease of use of a new automated device for blood glucose monitoring using the arm as an alternative sampling site. RESEARCH DESIGN AND METHODS: These studies use an automated hand-held device that applies a small vacuum, lances the skin, transfers blood onto an electrochemical test strip, and measures glucose. Patients who had type 1 or type 2 diabetes and had received no prior training using this device were recruited from five diabetes clinics. Testing was performed by the patients using this device and by trained healthcare professionals. Blood glucose was measured by 354 patients: from the arm using the device, from the finger using a laboratory reference instrument, and from the finger using the device via the secondary test port. Each patient completed a questionnaire rating the level of pain and ease of use of the device. RESULTS: Blood glucose results in samples obtained from the arm with the automated device agreed well with finger-stick plasma glucose results using a reference instrument (regression slope 0.98, intercept 0.01 mmol/l [0.1 mg/dl], r = 0.96). Error grid analysis showed that 100% of the measurements fell within zones A and B. In the survey, 60% of the patients reported that arm testing with the automated device was "painless;" another 31% of the patients stated that it was "much less painful," and 6% of patients considered using the device "less painful" than finger-stick testing. In a survey containing 15 questions for rating the ease of use with a scale of 1 to 6, the overall mean rating was 5.5. CONCLUSIONS: The automated device is easy to use and provides accurate glucose results; 97% of the patients found it less painful than finger-stick testing. PMID- 11423506 TI - Randomized dose ranging study of the reduction of fasting and postprandial glucose in type 2 diabetes by nateglinide (A-4166). AB - OBJECTIVE: This randomized crossover double-blind placebo-controlled study aimed to assess the efficacy of nateglinide (A-4166), a novel phenylalanine-derived insulin secretagogue, in type 2 diabetic subjects while fasting and 5 min before a standard meal. RESEARCH DESIGN AND METHODS: A single dose of nateglinide (60, 120, or 180 mg) or placebo was given to eight diet-treated overnight-fasted type 2 diabetic patients and to seven patients 5 min before a standard breakfast. Plasma glucose, radioimmunoassay insulin, and nateglinide were measured at baseline and for a further 180 min. RESULTS: The time-averaged 180-min postdose mean decrease in fasting plasma glucose concentration was greater after nateglinide (1.8 mmol/l; 95% CI 1.5-2.0) than after placebo (0.7 mmol/l; 95% CI 0.3-1.2) (P < 0.001). Hypoglycemia did not develop in any of the subjects. Insulin concentrations increased 1.5-, 1.8-, and 1.9-fold with the 60-, 120-, and 180-mg doses, respectively (P < 0.001), peaking approximately 30 min after the dose. Nateglinide concentrations peaked after approximately 30 min, decreasing to 21% of peak by 180 min. In the meal test, the mean increase (2.9 mmol/l, 2.3-3.6) in plasma glucose over 180 min after placebo was reduced by 1.8 mmol/l (P < 0.001) with the two higher doses of nateglinide. CONCLUSIONS: A single dose of nateglinide administered to diet-treated type 2 diabetic patients with fasting hyperglycemia increased insulin secretion and reduced fasting glucose without hypoglycemia. Administered 5 min before a meal, nateglinide reduced the postprandial glucose excursion by 64%. With its rapid onset and short duration of action, nateglinide is a promising oral prandial therapy in type 2 diabetes. PMID- 11423507 TI - A randomized trial of rosiglitazone therapy in patients with inadequately controlled insulin-treated type 2 diabetes. AB - OBJECTIVE: To determine the efficacy and safety of rosiglitazone (RSG) when added to insulin in the treatment of type 2 diabetic patients who are inadequately controlled on insulin monotherapy. RESEARCH DESIGN AND METHODS: After 8 weeks of insulin standardization and placebo (PBO) run-in, 319 type 2 diabetic patients with mean baseline HbA(1c) > or = 7.5% (8.9 +/- 1.1 to 9.1 +/- 1.3) on twice daily insulin therapy (total daily dose > or = 30 U) were randomized to 26 weeks of additional treatment with RSG (4 or 8 mg daily) or PBO. Insulin dose could be down- titrated only for safety reasons. The primary end point was reduction of HbA(1c) from baseline. RESULTS: RSG 4 and 8 mg daily significantly improved glycemic control, which was unchanged on PBO. By intent-to-treat analysis, treatment with RSG 8 mg plus insulin resulted in a mean reduction from baseline in HbA(1c) of 1.2% (P < 0.0001), despite a 12% mean reduction of insulin dosage. Over 50% of subjects treated daily with RSG 8 mg plus insulin had a reduction of HbA(1c) > or = 1.0%. Neither total:HDL cholesterol nor LDL:HDL cholesterol ratios significantly changed with RSG treatment. Serious adverse events did not differ among groups. CONCLUSIONS: The addition of RSG to insulin treatment results in significant improvement in glycemic control and is generally well tolerated. PMID- 11423508 TI - Prevalence of clinical and isolated subclinical cardiovascular disease in older adults with glucose disorders: the Cardiovascular Health Study. AB - OBJECTIVE: Clinical cardiovascular disease (CVD) is highly prevalent among people with diabetes. However, there is little information regarding the prevalence of subclinical CVD and its relation to clinical CVD in diabetes and in the glucose disorders that precede diabetes. RESEARCH DESIGN AND METHODS: Participants in the Cardiovascular Health Study, aged > or = 65 years (n = 5,888), underwent vascular and metabolic testing. Individuals with known disease in the coronary, cerebral, or peripheral circulations were considered to have clinical disease. Those without any clinical disease in whom CVD was detected by ultrasonography, electrocardiography, or ankle arm index in any of the three vascular beds were considered to have isolated subclinical disease. RESULTS: Approximately 30% of the cohort had clinical disease, and approximately 60% of the remainder had isolated subclinical disease. In those with normal glucose status, isolated subclinical disease made up most of the total CVD. With increasing glucose severity, the proportion of total CVD that was clinical disease increased; 75% of men and 66% of women with normal fasting glucose status had either clinical or subclinical CVD. Among those with known diabetes, the prevalence was approximately 88% (odds ratio [OR] 2.46 for men and 4.22 for women, P < 0.0001). There were intermediate prevalences and ORs for those with impaired fasting glucose status and newly diagnosed diabetes. CONCLUSIONS: Isolated subclinical CVD is common among older adults. Glucose disorders are associated with an increased prevalence of total CVD and an increased proportion of clinical disease relative to subclinical disease. PMID- 11423509 TI - Cross-sectional and prospective associations between proinsulin and cardiovascular disease risk factors in a population experiencing rapid cultural transition. AB - OBJECTIVE: To examine cross-sectional and prospective associations between proinsulin and cardiovascular disease risk factors using data from a population based study of type 2 diabetes among Native Canadians. RESEARCH DESIGN AND METHODS: Between 1993 and 1995, 72% of eligible members of a Native Canadian community participated in a baseline diabetes prevalence survey. Fasting samples were collected for glucose, C-peptide, proinsulin, lipids, and apolipoproteins. A 75-g oral glucose tolerance test was administered, and a second sample for glucose was drawn after 120 min. Blood pressure and waist circumference were determined. In the present study, subjects with normal glucose tolerance (NGT) (n = 505) and impaired glucose tolerance (IGT) (n = 74) were included in cross sectional analyses. In 1998, 95 individuals who had IGT or NGT at baseline with an elevated 2-h glucose concentration (> or = 7.0 mmol/l) participated in a follow-up evaluation using the protocol used at baseline. Cross-sectional and prospective associations between proinsulin and cardiovascular risk factors were assessed using correlation and multiple linear regression analyses. RESULTS: After adjustment for covariates including age, sex, C-peptide, waist circumference, and glucose tolerance status, fasting proinsulin concentration was significantly associated with concurrently measured lipid and apolipoprotein concentrations (triglycerides: r = 0.18, P < 0.0001; total cholesterol: r = 0.10, P = 0.02; LDL cholesterol: r = 0.11, P = 0.01; HDL cholesterol: r = -0.16, P = 0.0002; apolipoprotein (apo) B: r = 0.17, P < 0.0001; apoAI: r = -0.11, P = 0.008). In the adjusted prospective analysis, baseline triglycerides, HDL cholesterol, and apoB were associated with changes over time in proinsulin (r = 0.23, P = 0.04; r = -0.30, P = 0.01; r = 0.23, P = 0.04; respectively). CONCLUSIONS: These results confirm previously reported cross-sectional associations between proinsulin and lipid concentrations. In addition, an unexpected association between baseline lipids and proinsulin change was documented. PMID- 11423510 TI - Body fatness and fat distribution as predictors of metabolic abnormalities and early carotid atherosclerosis. AB - OBJECTIVE: To test the hypothesis that intra-abdominal fat plays a primary role over general adiposity for metabolic abnormalities and atherosclerosis. RESEARCH DESIGN AND METHODS: We cross-sectionally studied 849 Japanese men aged 50.3 +/- 8.5 years (range 20-78) with BMI 23.5 +/- 2.9 kg/m(2). Intimal-medial thickness (IMT) of the carotid artery was measured by ultrasound. General adiposity was assessed by BMI. Waist circumference and waist-to-hip ratio (WHR) were used as a surrogate measure for abdominal fat. Abdominal subcutaneous fat area (ASF) and intra-abdominal fat area (IAF) were measured by computed tomography. Correlations between these measures and carotid IMT were analyzed. The interaction of generalized adiposity (BMI) and IAF in relation to metabolic variables, such as glucose tolerance, insulin resistance, and serum lipids, was also evaluated. RESULTS: BMI, waist circumference, WHR, ASF, and IAF were all correlated with carotid IMT. Adjustment for BMI eliminated the associations between IMT and waist circumference, ASF, and IAF. In contrast, WHR retained a significant correlation with IMT. BMI and IAF were associated with insulin resistance, glucose tolerance, HDL cholesterol, and blood pressure independently of each other. IAF was an independent correlate for serum triglyceride, but BMI was not. CONCLUSIONS: The primary importance of IAF over general adiposity for carotid atherosclerosis was not confirmed. Caution is recommended when using WHR as a measure of abdominal fat. The roles of IAF for metabolic abnormalities may be more limited than conventionally thought. BMI and WHR are simple and better clinical predictors for carotid atherosclerosis versus IAF. PMID- 11423511 TI - Diminished insulin secretory response to glucose but normal insulin and glucagon secretory responses to arginine in a family with maternally inherited diabetes and deafness caused by mitochondrial tRNA(LEU(UUR)) gene mutation. AB - OBJECTIVE: The effects of glucose, arginine, and glucagon on beta-cell function as well as alpha-cell response to arginine were studied in a family with mitochondrial diabetes. RESEARCH DESIGN AND METHODS: The function of alpha- and beta-cells was assessed in all five siblings carrying the mitochondrial tRNA Leu(UUR) gene mutation at position 3243 and compared with six sex-, age-, and weight-matched control subjects. Insulin and C-peptide responses were evaluated by intravenous glucagon application, intravenous arginine stimulation test, and intravenous glucose tolerance test. Glucagon secretion was assessed during the arginine stimulation test. RESULTS: The glucose disappearance constant (K(g)) value (mean +/- SEM 0.61 +/- 0.04 vs. 1.1 +/- 0.04, P = 0.0002) as well as the acute insulin response to glucose (area under the curve [AUC] 0-10 min, 77.7 +/- 50.7 vs. 1,352.3 +/- 191.5 pmol/l, P = 0.0004) were decreased in all patients. Similarly, glucagon-stimulated C-peptide response was also impaired (728 +/- 111.4 vs. 1,526.7 +/- 157.7 pmol/l, P = 0.005), whereas the insulin response to arginine (AUC) was normal (1,346.9 +/- 710.8 vs. 1,083.2 +/- 132.5 pmol/l, P = 0.699). Acute glucagon response to arginine (AUC) was normal but tended to be higher in the patients than in the control subjects (181.7 +/- 47.5 vs. 90.0 +/- 21.1 pmol/l, P = 0.099). CONCLUSIONS: This study shows impaired insulin and C peptide secretion in response to a glucose challenge and to glucagon stimulation in diabetic patients with mitochondrial tRNA Leu(UUR) gene mutation, although insulin and glucagon secretory responses to arginine were normal. PMID- 11423512 TI - Amniotic fluid insulin at 14-20 weeks' gestation: association with later maternal glucose intolerance and birth macrosomia. AB - OBJECTIVE: To examine the hypothesis that early second trimester amniotic fluid (AF) insulin concentration is elevated and later fetal growth is augmented in gravidas demonstrating later oral glucose intolerance. RESEARCH DESIGN AND METHODS: In this prospective observational cohort study, AF was sampled at 14-20 weeks' gestation in 247 subjects, and 1-h 50-g oral glucose challenge tests (GCTs) were performed at > or = 24 weeks. AF insulin was assayed by an automated immuno-chemiluminometric assay (8). Macrosomia was defined as birth weight above the 90th centile. RESULTS: AF insulin concentration (range 1.4-44.5 pmol/l) correlated positively with gestational age and maternal weight. A logistic regression analysis, adjusted for maternal age and midpregnancy weight, showed increased AF insulin multiples of gestational age-specific medians to be associated with subsequently diagnosed gestational diabetes mellitus (GDM) (OR 1.9, CI 1.3-2.4, P = 0.029). Among 60 subjects with GCT values > 7.2 mmol/l, each unit increase in AF insulin multiple of median (MOM) was associated with a threefold increase in fetal macrosomia incidence (3.1, 1.3-4.9, P = 0.048). CONCLUSIONS: An elevated AF insulin concentration at 14-20 weeks' gestation is associated with subsequently documented maternal glucose intolerance. Among gravidas with GCT values > 7.2 mmol/l, elevated early AF insulin concentration is associated with fetal macrosomia. Maternal glucose intolerance may affect fetal insulin production before 20 weeks' gestation. PMID- 11423513 TI - Predictors of delayed gastric emptying in diabetes. AB - OBJECTIVE: To define the predictors of the rate of gastric emptying in patients with diabetes. RESEARCH DESIGN AND METHODS: A total of 101 outpatients with diabetes (79 type 1 and 22 type 2) underwent measurements of gastric emptying of a solid/liquid meal (scintigraphy), upper gastrointestinal symptoms (questionnaire), glycemic control (blood glucose concentrations during gastric emptying measurement), and autonomic nerve function (cardiovascular reflexes). RESULTS: The gastric emptying of solid and/or liquid was delayed in 66 (65%) patients. Solid (retention at 100 min 64 +/- 3.2 vs. 50.2 +/- 3.6%, P < 0.005) and liquid (retention at 100 min 22.7 +/- 1.7 vs. 16.0 +/- 1.8%, P < 0.001) gastric emptying was slower in women than in men. Of all upper gastrointestinal symptoms (including nausea and vomiting), only abdominal bloating/fullness was associated with slower gastric emptying (P < 0.005). A multiple regression analysis demonstrated that both abdominal bloating/fullness and female sex were predictors of slower gastric emptying of both solids and liquids. CONCLUSIONS: We conclude that the presence of abdominal bloating/fullness but not any other upper gastrointestinal symptom is associated with diabetic gastroparesis and that gastric emptying is slower in diabetic women than in diabetic men. PMID- 11423514 TI - Plantar tissue thickness is related to peak plantar pressure in the high-risk diabetic foot. AB - OBJECTIVE: To investigate the relationship among plantar foot pressure, plantar subcutaneous tissue thickness, severity of neuropathy (vibration perception threshold [VPT]), callus, and BMI in a large group of neuropathic diabetic patients at risk of foot ulceration. RESEARCH DESIGN AND METHODS: A total of 157 diabetic neuropathic patients (VPT >25 V) without either peripheral vascular or ulcer history were studied. Plantar foot pressure and plantar tissue thickness were measured at each metatarsal head (MTH) using an optical pedobarograph and an ultrasound scanning platform, respectively. RESULTS: A significant association was observed between peak plantar pressure and plantar tissue thickness at all MTHs (-0.26 < r < -0.61, P < 0.0001), with the least pronounced association at the first MTH. In addition, the pressure time integral was significantly associated with plantar tissue thickness (-0.24 < r < -0.57, P < 0.0001). BMI was significantly related to plantar tissue thickness (0.18 < r < 0.45, P < 0.05), but not to peak forefoot pressures. Subjects with callus had significantly reduced plantar tissue thickness at all MTHs except the first MTH and increased peak pressures at all MTHs (P < 0.001). CONCLUSIONS: This study confirms a strong inverse relationship between plantar tissue thickness and dynamic foot pressure measurements. Long-term follow-up of this patient population will confirm whether reduced plantar tissue thickness predicts the development of diabetic foot ulcers. PMID- 11423515 TI - Risk of developing retinopathy in Diabetes Control and Complications Trial type 1 diabetic patients with good or poor metabolic control. AB - OBJECTIVE: The study goal was to assess and predict the risk of developing retinopathy in type 1 diabetic patients with extreme metabolic control. RESEARCH DESIGN AND METHODS: Based on material from the Diabetes Control and Complications Trial (DCCT) study (n = 1,441 patients), patients without retinopathy at baseline (DCCT primary cohort) were considered under good or poor metabolic control if the mean HbA(1c) level (until the last visit) fell in the lower or upper 20% of the overall HbA(1c) distribution, respectively. Retinopathy was recorded as either absent or present. Logistic regression was used to predict retinopathy from covariates used in the DCCT retinopathy study. RESULTS: Among the 153 DCCT patients with "good metabolic control" (mean HbA(1c) < or = 6.87%), three-step change retinopathy developed in 15 (9.8%), and 138 (90%) remained free of retinopathy. Conversely, among the 166 patients with "poor metabolic control" (mean HbA(1c) > or = 9.49%), the complication did not develop in 71 (43%) and did develop in 95 (57%). Whereas occurrence of diabetic retinopathy was primarily due to metabolic control (P < 0.0001) and duration of participation in the study (P < 0.0001), two other covariates were found to be significant prognostic factors of the complication: HbA(1c) at baseline (OR 1.37, P < 0.001) and BMI (OR 1.11, P < 0.05). CONCLUSIONS: This study confirms that retinopathy develops in approximately 10% of patients with type 1 diabetes under good metabolic control, whereas > 40% of patients with type 1 diabetes remain free of retinopathy despite poor metabolic control. After adjusting for metabolic control and duration of participation in the study, it was found that previous glycemic exposure (HbA(1c)) and BMI may provide a possible explanation to such paradoxical clinical situations. PMID- 11423516 TI - The New York Diabetes Association's 48th Annual Scientific Meeting, the American Diabetes Association's 48th Annual Advanced Postgraduate Course, and the meeting of the Naomi Berry Diabetes Center of Columbia University. PMID- 11423517 TI - Psychosocial therapies in diabetes: report of the Psychosocial Therapies Working Group. AB - OBJECTIVE: To review key advances in the behavioral science literature related to psychosocial issues and therapies for persons with diabetes, to discuss barriers to research progress, and to make recommendations for future research. RESEARCH DESIGN AND METHODS: Key findings from the literature on psychosocial research in diabetes are reviewed separately for children and adults. Specific issues covered include psychosocial adjustment and psychiatric disorders, neurocognitive functioning, quality of life, and psychosocial therapies. Barriers that must be addressed to allow research in this area to progress are discussed. Recommendations are then made concerning high-priority areas for advancing research in the field. CONCLUSIONS: A substantial amount of behavioral science research has demonstrated that psychosocial factors play an integral role in the management of diabetes in both children and adults. Research has also shown the efficacy of a number of psychosocial therapies that can improve regimen adherence, glycemic control, psychosocial functioning, and quality of life. More research in this area is needed to develop psychosocial intervention programs for specific patient populations and to demonstrate the cost-effectiveness of these approaches. PMID- 11423518 TI - The concept of diabetes translation: addressing barriers to widespread adoption of new science into clinical care. PMID- 11423519 TI - Identification of the source of androgen excess in hyperandrogenic type 1 diabetic patients. PMID- 11423520 TI - Postprandial leptin responses after an oral fat tolerance test: differences in type 2 diabetes. PMID- 11423521 TI - The use of continuous insulin delivery systems in severely insulin-resistant patients. PMID- 11423522 TI - Metabolic control matters: Why is the message lost in the translation? The need for realistic goal-setting in diabetes care. PMID- 11423523 TI - Use of nicotinic acid in the management of recurrent hypoglycemic episodes in diabetes. PMID- 11423524 TI - Risky delay of hypoglycemia detection by glucose monitoring at the arm. PMID- 11423525 TI - High incidence of maternal transmission of diabetes in obese Uruguayan children. PMID- 11423528 TI - How cost-effective is the treatment of dyslipidemia in patients with diabetes but without cardiovascular disease? A response to Grover et al. PMID- 11423529 TI - Pseudonormal diastolic filling unmasked with glyceryl trinitrate in patients with type 2 diabetes with poor metabolic control. PMID- 11423531 TI - A protease-resistant prion protein isoform is present in urine of animals and humans affected with prion diseases. AB - Prion protein (PrP)(Sc), the only known component of the prion, is present mostly in the brains of animals and humans affected with prion diseases. We now show that a protease-resistant PrP isoform can also be detected in the urine of hamsters, cattle, and humans suffering from transmissible spongiform encephalopathies. Most important, this PrP isoform (UPrP(Sc)) was also found in the urine of hamsters inoculated with prions long before the appearance of clinical signs. Interestingly, intracerebrally inoculation of hamsters with UPrP(Sc) did not cause clinical signs of prion disease even after 270 days, suggesting it differs in its pathogenic properties from brain PrP(Sc). We propose that the detection of UPrP(Sc) can be used to diagnose humans and animals incubating prion diseases, as well as to increase our understanding on the metabolism of PrP(Sc) in vivo. PMID- 11423532 TI - Association of insulin-like growth factor 1 receptor with EHD1 and SNAP29. AB - Ligand-induced receptor-mediated endocytosis plays a central role in regulating signaling conveyed by tyrosine kinase receptors. This process depends on the recruitment of the adaptor protein 2 (AP-2) complex, clathrin, dynamin, and other accessory proteins to the ligand-bound receptor. We show here that besides AP-2 and clathrin, two other proteins participate in the endocytic process of the insulin-like growth factor receptor (IGF-1R); they are EHD1, an Eps15 homology (EH) domain-containing protein 1, and SNAP29, a synaptosomal-associated protein. EHD1 and SNAP29 form complexes with alpha-adaptin of AP-2 and co-localize in endocytic vesicles, indicating a role for them in endocytosis. EHD1 and SNAP29 interact directly with each other and are present in complexes with IGF-1R. After IGF-1 induction, EHD1 and IGF-1R co-localize intracellularly. Overexpression of EHD1 in Chinese hamster ovary cells represses IGF-1-mediated signaling, as measured by mitogen-activated protein kinase phosphorylation and Akt phosphorylation, indicating that EHD1 plays a role as a down-regulator in IGF-1 signaling pathway. PMID- 11423533 TI - Structure and expression of the Ah receptor repressor gene. AB - The aryl hydrocarbon receptor (AhR) repressor (AhRR) gene has been isolated and characterized from a mouse genomic library. The gene is distributed as 11 exons in a total length of about 60 kilobase pairs. Fluorescence in situ hybridization analysis has shown that the AhRR gene is located at mouse chromosome 13C2, at rat chromosome 1p11.2, and at human chromosome 5p15.3. The AhRR gene has a TATA-less promoter and several transcription start sites. In addition, putative regulatory DNA sequences such as xenobiotic responsive element (XRE), GC box, and NF-kappaB binding sites have been identified in the 5'-upstream region of the AhRR gene. Transient transfection analyses of HeLa cells with reporter genes that contain deletions and point mutations in the AhRR promoter revealed that all three XREs mediated the inducible expression of the AhRR gene by 3-methylcholanthrene treatment, and furthermore, GC box sequences were indispensable for a high level of inducible expression and for constitutive expression. Moreover, by using gel mobility shift assays we were able to show that the AhR/Arnt heterodimer binds to the XREs with very low affinity, which is due to three varied nucleotides outside the XRE core sequence. We have also shown that Sp1 and Sp3 can bind to the GC boxes. Finally, both transient transfection analysis and gel mobility shift assay revealed that the AhRR gene is up-regulated by a p65/p50 heterodimer that binds to the NF-kappaB site when the cells has been exposed to 12-O tetradecanoylphorbol-13-acetate, and this inducible expression was further enhanced by cotreatment of 12-O-tetradecanoylphorbol-13-acetate and 3 methylcholanthrene. PMID- 11423534 TI - Identification and functional analysis of splice variants of the germ cell soluble adenylyl cyclase. AB - In mammalian germ cells, cAMP signaling is dependent on two forms of adenylyl cyclase, the conventional membrane-bound ACIII and a soluble form of adenylyl cyclase (sAC). Recent elucidation of the sAC sequence indicates that this enzyme is phylogenetically distinct from the membrane-bound AC, does not interact with G proteins, and its activity is regulated by bicarbonate ions. Here we have investigated the properties and regulation of this enzyme during spermatogenesis. Two different transcripts encoding a full-length and truncated sAC were identified by reverse transcriptase-polymerase chain reaction and RNase protection analysis. The truncated sAC transcript lacks exon 11 with a premature termination of the open reading frame after the catalytic domain. Reverse transcriptase-polymerase chain reaction with testis RNA from adult mouse and rat of different ages, as well as RNase protection, showed that both transcripts are absent at 11 days of age, appear between 20 and 30 days of age, and are retained in the adult testis. The presence of corresponding proteins in testis, germ cells, and spermatozoa was demonstrated by fast protein liquid chromatography and differential immunoprecipitation with full-length sAC-specific antibodies. Bicarbonate ions activated both sAC forms and increased cAMP levels in germ cells isolated from 25- and 50-day-old rats and adult rats in a concentration-dependent manner. These findings provide evidence that full-length and truncated sAC are generated by alternate splicing. Both forms are active in spermatids, and the bicarbonate present in the seminiferous tubule may be a signal that regulates cAMP levels in these cells. PMID- 11423535 TI - L-cysteine-mediated destabilization of dinitrosyl iron complexes in proteins. AB - Nitric oxide is a signaling molecule in intercellular communication as well as a powerful weapon used by macrophages to kill tumor cells and pathogenic bacteria. Here, we show that when Escherichia coli cells are exposed to nitric oxide, its ferredoxin [2Fe-2S] cluster is nitrosylated, forming the dinitrosyl iron complex with a characteristic EPR signal at g(av) = 2.04. Such formed ferredoxin dinitrosyl iron complex is efficiently repaired in E. coli cells even in the absence of new protein synthesis. However, the repair activity is completely inactivated once E. coli cells are disrupted, indicating that repairing the ferredoxin dinitrosyl iron complex requires cellular reducing equivalents. In search of such cellular factors, we find that l-cysteine can effectively eliminate the EPR signal of the ferredoxin dinitrosyl iron complex and release the ferrous iron from the complex. In contrast, N-acetyl-l-cysteine and reduced glutathione are much less effective. l-Cysteine seems to have a general function, since it can also remove the otherwise stable dinitrosyl iron complexes from proteins in the cell extracts prepared from the E. coli cells treated with nitric oxide. We propose that l-cysteine is responsible for removing the dinitrosyl iron complexes from the nitric oxide-modified proteins into which a new iron-sulfur cluster will be reassembled. PMID- 11423536 TI - Solution structure and interaction with basic and acidic fibroblast growth factor of a 3-kDa human platelet factor-4 fragment with antiangiogenic activity. AB - Platelet factor-4 is a protein belonging to the family of ELR-negative CXC chemokines which binds to fibroblast growth factor and inhibits its mitogenic activity. Platelet factor-4 also inhibits tumor growth by mechanisms involving antiangiogenesis. Antiangiogenic activity in vitro has also been shown for the 24 residue C-terminal fragment of the protein, which decreases the affinity between basic fibroblast growth factor and its cell-surface receptor. In this study, the preferential conformation of this fragment in solution has been determined and has been found to be composed of two helical subdomains. In addition, we show that the fragment forms a specific 1:1 complex with acidic and basic fibroblast growth factors and that both subdomains are probably required for inhibition of fibroblast growth factor-driven mitogenesis. Finally, we show that the binding of the fragment alters the structure of the fibroblast growth factors, although some of such alterations do not seem related with the inhibition of mitogenic activity. Since this fragment has recently been shown to inhibit fibroblast growth factor-induced angiogenesis in vivo when injected intraperitoneally, these results are relevant for developing new antiangiogenic treatments. PMID- 11423537 TI - Human ABCA1 BAC transgenic mice show increased high density lipoprotein cholesterol and ApoAI-dependent efflux stimulated by an internal promoter containing liver X receptor response elements in intron 1. AB - By using BAC transgenic mice, we have shown that increased human ABCA1 protein expression results in a significant increase in cholesterol efflux in different tissues and marked elevation in high density lipoprotein (HDL)-cholesterol levels associated with increases in apoAI and apoAII. Three novel ABCA1 transcripts containing three different transcription initiation sites that utilize sequences in intron 1 have been identified. In BAC transgenic mice there is an increased expression of ABCA1 protein, but the distribution of the ABCA1 product in different cells remains similar to wild type mice. An internal promoter in human intron 1 containing liver X response elements is functional in vivo and directly contributes to regulation of the human ABCA1 gene in multiple tissues and to raised HDL cholesterol, apoAI, and apoAII levels. A highly significant relationship between raised protein levels, increased efflux, and level of HDL elevation is evident. These data provide proof of the principle that increased human ABCA1 efflux activity is associated with an increase in HDL levels in vivo. PMID- 11423538 TI - Disruption of thylakoid-associated kinase 1 leads to alteration of light harvesting in Arabidopsis. AB - To survive fluctuations in quality and intensity of light, plants and algae are able to preferentially direct the absorption of light energy to either one of the two photosystems, PSI or PSII. This rapid process is referred to as a state transition and has been correlated with the phosphorylation and migration of the light-harvesting complex protein (LHCP) between PSII and PSI. We show here that thylakoid protein kinases (TAKs) are required for state transitions in Arabidopsis. Antisense TAK1 expression leads to a loss of LHCP phosphorylation and a reduction in state transitions. Preferential activation of PSII causes LHCP to accumulate with PSI, and TAK1 mutants disrupt this process. Finally, TAKs also influence the phosphorylation of multiple thylakoid proteins. PMID- 11423539 TI - A non-Golgi alpha 1,2-fucosyltransferase that modifies Skp1 in the cytoplasm of Dictyostelium. AB - Skp1 is a subunit of the SCF-E3 ubiquitin ligase that targets cell cycle and other regulatory factors for degradation. In Dictyostelium, Skp1 is modified by a pentasaccharide containing the type 1 blood group H trisaccharide at its core. To address how the third sugar, fucose alpha1,2-linked to galactose, is attached, a proteomics strategy was applied to determine the primary structure of FT85, previously shown to copurify with the GDP-Fuc:Skp1 alpha 1,2-fucosyltransferase. Tryptic-generated peptides of FT85 were sequenced de novo using Q-TOF tandem mass spectrometry. Degenerate primers were used to amplify FT85 genomic DNA, which was further extended by a novel linker polymerase chain reaction method to yield an intronless open reading frame of 768 amino acids. Disruption of the FT85 gene by homologous recombination resulted in viable cells, which had altered light scattering properties as revealed by flow cytometry. FT85 was necessary and sufficient for Skp1 fucosylation, based on biochemical analysis of FT85 mutant cells and Escherichia coli that express FT85 recombinantly. FT85 lacks sequence motifs that characterize all other known alpha 1,2-fucosyltransferases and lacks the signal-anchor sequence that targets them to the secretory pathway. The C terminal region of FT85 harbors motifs found in inverting Family 2 glycosyltransferase domains, and its expression in FT85 mutant cells restores fucosyltransferase activity toward a simple disaccharide substrate. Whereas most prokaryote and eukaryote Family 2 glycosyltransferases are membrane-bound and oriented toward the cytoplasm where they glycosylate lipid-linked or polysaccharide precursors prior to membrane translocation, the soluble, eukaryotic Skp1-fucosyltransferase modifies a protein that resides in the cytoplasm and nucleus. PMID- 11423540 TI - Cytoprotection against mechanical forces delivered through beta 1 integrins requires induction of filamin A. AB - Cells in mechanically active environments can activate cytoprotective mechanisms to maintain membrane integrity in the face of potentially lethal applied forces. Cytoprotection may be mediated by expression of membrane-associated cytoskeletal proteins including filamin A, an actin-binding protein that increases the rigidity of the subcortical actin cytoskeleton. In this study, we tested the hypotheses that applied forces induce the expression of filamin A specifically and that this putative protective response inhibits cell death. Magnetically generated forces were applied to protein-coated magnetite beads bound to human gingival fibroblasts, cells with constitutively low basal levels of filamin A mRNA and protein. Forces applied through collagen or fibronectin, but not bovine serum albumin or poly-l-lysine-coated beads, increased mRNA and protein content of filamin A by 3-7-fold. Forces had no effect on the expression of other filamin isotypes or other cytoskeletal proteins. This effect was dependent on the duration of force and was blocked by anti-beta(1) integrin antibodies. Force also stimulated a 60% increase in expression of luciferase under the control of a filamin A promoter in transiently transfected Rat2 fibroblasts and was dependent on Sp1 transcription factor binding sites located immediately upstream of the transcription start site. Experiments with actinomycin D-treated cells showed that the increased filamin A expression after force application was due in part to prolongation of mRNA half-life. Antisense filamin oligonucleotides blocked force-induced filamin A expression and increased cell death by >2-fold above controls. The force-induced regulation of filamin A was dependent on intact actin filaments. We conclude that cells from mechanically active environments can couple diverse signals from forces applied through beta-integrins to up-regulate the production of cytoprotective cytoskeletal proteins, typified by filamin A. PMID- 11423541 TI - Alanine metabolism in the perfused rat liver. Studies with (15)N. AB - We have utilized [(15)N]alanine or (15)NH(3) as metabolic tracers in order to identify sources of nitrogen for hepatic ureagenesis in a liver perfusion system. Studies were done in the presence and absence of physiologic concentrations of portal venous ammonia in order to test the hypothesis that, when the NH(4)(+):aspartate ratio is >1, increased hepatic proteolysis provides cytoplasmic aspartate in order to support ureagenesis. When 1 mm [(15)N]alanine was the sole nitrogen source, the amino group was incorporated into both nitrogens of urea and both nitrogens of glutamine. However, when studies were done with 1 mm alanine and 0.3 mm NH(4)Cl, alanine failed to provide aspartate at a rate that would have detoxified all administered ammonia. Under these circumstances, the presence of ammonia at a physiologic concentration stimulated hepatic proteolysis. In perfusions with alanine alone, approximately 400 nmol of nitrogen/min/g liver was needed to satisfy the balance between nitrogen intake and nitrogen output. When the model included alanine and NH(4)Cl, 1000 nmol of nitrogen/min/g liver were formed from an intra-hepatic source, presumably proteolysis. In this manner, the internal pool provided the cytoplasmic aspartate that allowed the liver to dispose of mitochondrial carbamyl phosphate that was rapidly produced from external ammonia. This information may be relevant to those clinical situations (renal failure, cirrhosis, starvation, low protein diet, and malignancy) when portal venous NH(4)(+) greatly exceeds the concentration of aspartate. Under these circumstances, the liver must summon internal pools of protein in order to accommodate the ammonia burden. PMID- 11423542 TI - Distinct ligand-binding modes for integrin alpha(v)beta(3)-mediated adhesion to fibronectin versus vitronectin. AB - Cell surface integrins can adopt distinct conformations in response to ligand binding and intracellular signals. Several integrins including alpha(v)beta(3) can bind to multiple ligands. The binding of alpha(v)beta(3) to fibronectin and vitronectin was used as a model to determine whether the same or distinct forms of the receptor were utilized in strong binding to the two different ligands. A spinning-disc device was used to measure the relative strength of the alpha(v)beta(3)-ligand bonds. The initial binding reaction for both ligands occurred in the absence of metabolic energy and resulted in a strong adhesion to fibronectin but a weak adhesion to vitronectin. Increases in the strength of the alpha(v)beta(3)-vitronectin bond required phosphorylation of the beta(3) cytoplasmic domain, intracellular signals, and the binding of cytoskeletal proteins to cytoplasmic domains of beta(3) controlled by Tyr-747 and Tyr-759. In contrast, alpha(v)beta(3)-mediated adhesion to fibronectin was unaffected by phorbol 12-myristate 13-acetate, mutations of Tyr-747 and Tyr-759 to phenylalanine, or availability of metabolic energy. This suggests that strong adhesion to fibronectin used the initial binding conformation, whereas strong binding to vitronectin required signaling-induced changes in the conformation of alpha(v)beta(3). PMID- 11423543 TI - Heterogeneous fatty acylation of Src family kinases with polyunsaturated fatty acids regulates raft localization and signal transduction. AB - Fatty acylation of Src family kinases is essential for localization of the modified proteins to the plasma membrane and to plasma membrane rafts. It has been suggested that the presence of saturated fatty acyl chains on proteins is conducive for their insertion into liquid ordered lipid domains present in rafts. The ability of unsaturated dietary fatty acids to be attached to Src family kinases has not been investigated. Here we demonstrate that heterogeneous fatty acylation of Src family kinases occurs and that the nature of the attached fatty acid influences raft-mediated signal transduction. By using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, we show that in addition to 14:0 (myristate), 14:1 and 14:2 fatty acids can be attached to the N-terminal glycine of the Src family kinase Fyn when the growth media are supplemented with these dietary fatty acids. Moreover, we synthesized novel iodinated analogs of oleate and stearate, and we showed that heterogeneous S-acylation can occur on cysteine residues within Fyn as well as Galpha, GAP43, and Ras. Modification of Fyn with unsaturated or polyunsaturated fatty acids reduced its raft localization and resulted in decreased T cell signal transduction. These studies establish that heterogeneous fatty acylation is a widespread occurrence that serves to regulate signal transduction by membrane-bound proteins. PMID- 11423544 TI - Adenosine 5'-monophosphate inhibits the association of 14-3-3 proteins with the plant plasma membrane H(+)-ATPase. AB - Although a well ascertained evidence proves that the activity of the plant plasma membrane H(+)-ATPase is regulated by 14-3-3 proteins, information about physiological factors modulating the phosphorylation-dependent association between 14-3-3 proteins and the proton pump is largely incomplete. In this paper we show that the 5'-AMP-mimetic, 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR), inhibits the fusicoccin-promoted proton extrusion in maize roots. We also demonstrate that 5'-AMP inhibits the association of 14-3-3 proteins with the C-terminal domain of the H(+)-ATPase in an overlay assay as well as the 14-3-3 dependent stimulation of the Arabidopsis thaliana H(+)-ATPase AHA1 isoform expressed in yeast membranes. Finally, by means of affinity chromatography with immobilized 5'-AMP and trinitrophenyl-AMP fluorescence analysis, we demonstrate that the 14-3-3 isoform GF14-6 from maize is able to bind 5'-AMP. The possible role of 5'-AMP as a general regulator of 14-3-3 functions in the plant cell is discussed. PMID- 11423545 TI - Differential induction of Hsp70-encoding genes in human hematopoietic cells. AB - The rapid transcriptional activation of heat shock genes in response to stress is crucial for the cellular survival and the development of thermotolerance. Although heat shock response is a widespread phenomenon, certain cells exhibit a diminished induction of heat shock gene expression upon stress stimuli. Here we have analyzed the development of thermotolerance and induction of distinct Hsp70 encoding genes in three cell lines representing different hematopoietic cell types. We show that in response to heat shock, cell survival and induction of thermotolerance are impaired in Raji and HL60 cells, as compared with K562 cells. Accordingly, transcriptional induction of the hsp70 gene is diminished in Raji and HL60 cells. This appears to be due to inability of transcription factors, including HSF1 to bind to the hsp70.1 promoter in vivo. Consistent with the genomic footprint, analysis of hsp70.1 mRNA expression using a specific 3' untranslated region probe reveals that induction of the hsp70.1 gene upon heat shock is completely abolished in Raji and HL60 cells. The suppression of the hsp70.1 promoter is not caused by impaired function of HSF1, since HSF1 is equally activated in all cell types and occupies another heat-inducible promoter, hsp90 alpha. Furthermore, among distinct inducible hsp70 genes, suppression seems to be specific for the hsp70.1 gene, since heat shock results in induction of hsp70.2 and hsp70B' mRNA expression in all cell lines. Taken together, our results demonstrate that distinct Hsp70-encoding genes contribute to the heat shock response in a cell type-dependent manner. PMID- 11423546 TI - Investigations of the in vitro transport of human milk oligosaccharides by a Caco 2 monolayer using a novel high performance liquid chromatography-mass spectrometry technique. AB - Complex lactose-derived oligosaccharides belong to the main components of human milk and are believed to exert multiple functions in the breast-fed infant. Therefore, we investigated the transepithelial transport of human milk oligosaccharides over Caco-2 monolayers. Main human milk oligosaccharides (HMOs) in the apical, basolateral, or intracellular compartment were separated by high performance liquid chromatography using a Hypercarb(TM) column and analyzed on line by mass spectrometry. This method allowed the identification and quantification of these components in intra- and extracellular fractions without prior purification. Using this technique we were able to show that acidic and neutral HMOs cross the epithelial barrier. The transepithelial flux of neutral, but not acidic, oligosaccharides was temperature-sensitive and partly inhibited by brefeldin A and bafilomycin A. Furthermore, net flux from the apical to the basolateral compartment was only observed for the neutral components. Similarly, apical cellular uptake was only found for neutral components but not for acidic oligosaccharides. Intracellular concentrations of neutral HMOs were significantly increased by inhibitors of transcytosis such as brefeldin A, N-ethylmaleimide, or bafilomycin A. The cellular uptake was saturable, and an apparent K(m) for lacto N-fucopentaose I of 1.7 +/- 0.1 mmol/liter and for lacto-N-tetraose of 1.8 +/- 0.4 mmol/liter was determined. Furthermore, the uptake of lacto-N-fucopentaose I could be inhibited by the addition of the stereoisomer lacto-N-fucopentaose II but not by lacto-N-tetraose. These findings suggest that neutral HMOs are transported across the intestinal epithelium by receptor-mediated transcytosis as well as via paracellular pathways, whereas translocation of acidic HMOs solely represents paracellular flux. PMID- 11423547 TI - New insights on how metals disrupt amyloid beta-aggregation and their effects on amyloid-beta cytotoxicity. AB - Amyloid-beta protein (A beta) aggregates in the brain to form senile plaques. By using thioflavin T, a dye that specifically binds to fibrillar structures, we found that metals such as Zn(II) and Cu(II) normally inhibit amyloid beta aggregation. Another method for detecting A beta, which does not distinguish the types of aggregates, showed that these metals induce a non-beta-sheeted aggregation, as reported previously. Secondary structural analysis and microscopic studies revealed that metals induced A beta to make non-fibrillar aggregates by disrupting beta-sheet formation. These non-fibrillar A beta aggregates displayed much weaker Congo Red birefringence, and in separate cell culture experiments, were less toxic than self beta-aggregates, as demonstrated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The toxicity of soluble A beta was enhanced in the presence of Cu(II), which suggests the previously hypothesized role of A beta in generating oxidative stress. Finally, under an acidic condition, similar to that in the inflammation associated with senile plaques, beta-aggregation was robustly facilitated at one specific concentration of Zn(II) in the presence of heparin. However, because a higher concentration of Zn(II) virtually abolished this abnormal phenomenon, and at normal pH any concentrations strongly inhibit beta-aggregation and its associated cytotoxicity, including its anti-oxidative nature we suggest that Zn(II) has an overall protective effect against beta-amyloid toxicity. PMID- 11423548 TI - Characterization of a general stabilizer element that blocks deadenylation dependent mRNA decay. AB - mRNA degradation is a regulated process that can play an important role in determining the level of expression of specific genes. The rate at which a specific mRNA is degraded depends largely on specific cis-acting sequences located throughout the transcript. cis-Acting destabilizer sequences that promote increased rates of decay have been identified in several short-lived mRNAs. However, little is known about elements that promote stability, known as stabilizer elements (STEs), and how they function. The work presented here describes the characterization of a STE in the PGK1 transcript. The PGK1 stabilizer element (P-STE) has been delineated to a 64-nucleotide sequence from the coding region that can stabilize a chimeric transcript containing the instability elements from the 3'-untranslated region of the MFA2 transcript. The P-STE is located within the PGK1 coding region and functions when located in the translated portion of the transcript and at a minimum distance from the 3' untranslated region. These results further support the link between translation and mRNA degradation. A conserved sequence in the TEF1/2 transcript has been identified that also functions as a STE, suggesting that this sequence element maybe a general stability determinant found in other yeast mRNAs. PMID- 11423549 TI - Analysis of the alpha-actinin/zyxin interaction. AB - The yeast two-hybrid system was used to search for interaction partners of human zyxin. Screening of two different cDNA libraries, one prepared from human placenta, the other from human heart, yielded several positive clones that occurred in both searches, including clones coding for cyclophilin, nebulette, and alpha-actinin. The zyxin/alpha-actinin interaction was analyzed in detail. By site-directed mutagenesis, a linear motif of 6 amino acids (Phe-Gly-Pro-Val-Val Ala) present at the N terminus of zyxin was found to play a critical role. Replacement of a single amino acid within this motif abolished binding to alpha actinin in blot overlays as well as in living cells. On the other hand, the interaction site in alpha-actinin was mapped to a conformational determinant present in the center of the protein as demonstrated by a fragment deletion analysis. This binding site involved a tandem array of two complete spectrin-like domains. Only fragments that were able to dimerize in yeast also bound to zyxin, suggesting that dimerization of alpha-actinin is essential for zyxin binding. PMID- 11423550 TI - Structural and functional characterization of protein 4.1R-phosphatidylserine interaction: potential role in 4.1R sorting within cells. AB - Erythrocyte protein 4.1R is a multifunctional protein that binds to various membrane proteins and to phosphatidylserine. In the present study, we report two important observations concerning 4.1R-phosphatidylserine interaction. Biochemically, a major finding of the present study is that 4.1R binding to phosphatidylserine appears to be a two-step process in which 4.1R first interacts with serine head group of phosphatidylserine through the positively charged amino acids YKRS and subsequently forms a tight hydrophobic interaction with fatty acid moieties. 4.1R failed to dissociate from phosphatidylserine liposomes under high ionic strength but could be released specifically by phospholipase A(2) but not by phospholipase C or D. Biochemical analyses showed that acyl chains were associated with 4.1R released by phospholipase A(2). Importantly, the association of acyl chains with 4.1R impaired its ability to interact with calmodulin, band 3, and glycophorin C. Removal of acyl chains restored 4.1R binding. These data indicate that acyl chains of phosphatidylserine play an important role in its interaction with 4.1R and on 4.1R function. In terms of biological significance, we have obtained evidence that 4.1R-phosphatidylserine interaction may play an important role in cellular sorting of 4.1R. PMID- 11423551 TI - MAP kinase phosphatase-1 gene transcription in rat neuroendocrine cells is modulated by a calcium-sensitive block to elongation in the first exon. AB - Transcriptional elongation of many eukaryotic, prokaryotic, and viral genes is tightly controlled, which contributes to gene regulation. Here we describe this phenomenon for the MAP kinase phosphatase 1 (MKP-1) immediate early gene. In rat GH4C1 pituitary cells, MKP-1 mRNA is rapidly and transiently induced by the thyrotropin-releasing hormone (TRH) and the epidermal growth factor EGF via transcriptional activation of the gene. Ca(2+) signals are necessary for the induction of MKP-1 in response to TRH but not to EGF. Reporter gene analysis with the newly cloned rat promoter sequence shows only limited induction in response to various stimuli, including TRH or EGF. By nuclear run-on assays we demonstrate that in basal conditions, a strong block to elongation in the first exon regulates the MKP-1 gene and that stimulation with either TRH or EGF overcomes the block. Ca(2+) signals are important to release the MKP-1 elongation block in a manner similar to the c-fos oncogene. These results suggest that a common mechanism of intragenic regulation may be conserved between MKP-1 and c-fos in mammalian cells. PMID- 11423552 TI - The transcription factors SOX9 and SOX10 are vitiligo autoantigens in autoimmune polyendocrine syndrome type I. AB - Vitiligo is common in the hereditary disorder autoimmune polyendocrine syndrome type I (APS I). Patients with APS I are known to have high titer autoantibodies directed against various tissue-specific antigens. Using sera from APS I patients for immunoscreening of a cDNA library from human scalp, we identified the transcription factors SOX9 and SOX10 as novel autoantigens related to this syndrome. Immunoreactivity against SOX9 was found in 14 (15%) and against SOX10 in 20 (22%) of the 91 APS I sera studied. All patients reacting with SOX9 displayed reactivity against SOX10, suggesting shared epitopes. Among the 19 patients with vitiligo, 12 (63%) were positive for SOX10 (p < 0.0001). Furthermore, three of 93 sera from patients with vitiligo unrelated to APS I showed strong reactivity against SOX10, which may indicate a more general role of SOX10 as an autoantigen in vitiligo. PMID- 11423553 TI - Cloning, expression, and characterization of tomato (Lycopersicon esculentum) aminopeptidase P. AB - A cDNA (LeAPP2) was cloned from tomato coding for a 654 amino acid protein of 72.7 kDa. The deduced amino acid sequence was >40% identical with that of mammalian aminopeptidase P, a metalloexopeptidase. All amino acids reported to be important for binding of the active site metals and catalytic activity, respectively, were conserved between LeAPP2 and its mammalian homologues. LeAPP2 was expressed in Escherichia coli in N-terminal fusion with glutathione S transferase and was purified from bacterial extracts. LeAPP2 was verified as an aminopeptidase P, hydrolyzing the amino-terminal Xaa-Pro bonds of bradykinin and substance P. LeAPP2 also exhibited endoproteolytic activity cleaving, albeit at a reduced rate, the internal -Phe-Gly bond of substance P. Apparent K(m) (15.2 +/- 2.4 microm) and K(m)/k(cat) (0.94 +/- 0.11 mm(-1) x s(-1)) values were obtained for H-Lys(Abz)-Pro-Pro-pNA as the substrate. LeAPP2 activity was maximally stimulated by addition of 4 mm MnCl(2) and to some extent also by Mg(2+), Ca(2+), and Co(2+), whereas other divalent metal ions (Cu(2+), Zn(2+)) were inhibitory. Chelating agents and thiol-modifying reagents inhibited the enzyme. The data are consistent with LeAPP2 being a Mn(II)-dependent metalloprotease. This is the first characterization of a plant aminopeptidase P. PMID- 11423554 TI - Membrane binding and substrate access merge in cytochrome P450 7A1, a key enzyme in degradation of cholesterol. AB - To study membrane topology and mechanism for substrate specificity, we truncated residues 2-24 in microsomal cytochrome P450 7A1 (P450 7A1) and introduced conservative and nonconservative substitutions at positions 214-227. Heterologous expression in Escherichia coli was followed by investigation of the subcellular distribution of the mutant P450s and determination of the kinetic and substrate binding parameters for cholesterol. The results indicate that a hydrophobic region, comprising residues 214-227, forms a secondary site of attachment to the membrane in P450 7A1 in addition to the NH(2)-terminal signal-anchor sequence. There are two groups of residues at this enzyme-membrane interface. The first are those whose mutation results in more cytosolic P450 (Val-214, His-225, and Met 226). The second group are those whose mutation leads to more membrane-bound P450 (Phe-215, Leu-218, Ile-224, and Phe-227). In addition, the V214A, V214L, V214T, F215A, F215L, F215Y, L218I, L218V, V219T, and M226A mutants showed a 5-12-fold increased K(m) for cholesterol. The k(cat) of the V214A, V214L, V219T, and M226A mutants was increased up to 1.8-fold, and that of the V214T, F215A, F215L, F215Y, L218I, and L218V mutants was decreased 3-10.5-fold. Based on analysis of these mutations we suggest that cholesterol enters P450 7A1 through the membrane, and Val-214, Phe-215, and Leu-218 are the residues located near the point of cholesterol entry. The results provide an understanding of both the P450 7A1 membrane interactions and the mechanism for substrate specificity. PMID- 11423555 TI - Prostaglandin E(2) regulates the level and stability of cyclooxygenase-2 mRNA through activation of p38 mitogen-activated protein kinase in interleukin-1 beta treated human synovial fibroblasts. AB - The p38 MAPK mediates transcriptional and post-transcriptional control of cyclooxygenase-2 (COX-2) mRNA following interleukin-1(IL-1)/lipopolysaccharide cellular activation. We explored a positive feedback, prostaglandin E(2) (PGE(2)) dependent stabilization of COX-2 mRNA mediated by the p38 MAPK cascade in IL-1 beta-stimulated human synovial fibroblasts. We observed a rapid (5 min), massive (>30-fold), and sustained (>48 h) increase in COX-2 mRNA, protein, and PGE(2) release following a recombinant human (rh) IL-1 beta signal that was inhibited by NS-398, a COX-2 inhibitor, and SB202190, a selective, cell-permeable p38 MAPK inhibitor. PGE(2) completely reversed NS-398-mediated inhibition but not SB202190 dependent inhibition. The eicosanoid didn't potentiate IL-1 beta-induced COX-2 expression nor did it activate COX-2 gene expression in quiescent cells. Transfection experiments with a human COX-2 promoter construct revealed a minor element of p38 MAPK-dependent transcriptional control after IL-1 beta stimulation. p38 MAPK synergized with the cAMP/cAMP-dependent protein kinase cascade to transactivate the COX-2 promoter. When human synovial fibroblasts were activated with rhIL-1 beta for 3-4 h (steady state) followed by washout, the elevated levels of COX-2 mRNA declined rapidly (<2 h) to control levels. If PGE(2), unlike EP2/3 agonists butaprost and sulprostone, was added to fresh medium, COX-2 mRNA levels remained elevated for up to 16 h. SB202190 or anti PGE(2) monoclonal antibody compromised the stabilization of COX-2 mRNA by PGE(2). Deletion analysis using transfected chimeric luciferase-COX-2 mRNA 3' untranslated region reporter constructs revealed that IL-1 beta increased reporter gene mRNA stability and translation via AU-containing distal regions of the untranslated region. This response was mediated entirely by a PGE(2)/p38 MAPK dependent process. We conclude that the magnitude and duration of the induction of COX-2 mRNA, protein, and PGE(2) release by rhIL-1 beta is primarily the result of PGE(2)-dependent stabilization of COX-2 mRNA and stimulation of translation, a process involving a positive feedback loop mediated by the EP4 receptor and the downstream kinases p38 MAPK and, perhaps, cAMP-dependent protein kinase. PMID- 11423556 TI - The folding and stability of human alpha class glutathione transferase A1-1 depend on distinct roles of a conserved N-capping box and hydrophobic staple motif. AB - An N-capping box and a hydrophobic staple motif are strictly conserved in the core of all known glutathione S-transferases (GST). In the present work, mutations of hGSTA1-1 enzyme residues forming these motifs have been generated. The analysis of S154A, D157A, and S154A/D157A capping mutants indicate that the removal of this local signal destabilizes the protein. The fact that the third helical residue D157A mutation (N-3) was much more destabilizing than the first helical residue S154A mutation (N-cap) suggests that the appropriate conformation of the conserved substructure formed by the alpha 6-helix and preceding loop (GST motif II) is crucial for the overall protein stability. The refolding study of GSTA1-1 variants supports the prediction that this subdomain could represent a nucleation site of refolding. The analysis of L153A, I158A, L153G, and L153A/I158A hydrophobic staple mutants indicate that the removal of this motif destabilizes the GSTA1-1 structure as well as its refolding transition state. The hydrophobic staple interaction favors essential inter-domain contacts and, thereby, in contrast to capping interactions, accelerates the enzyme reactivation. Its strict conservation in the GST system supports the suggestion that this local signal could represent an evolutionarily conserved determinant for rapid folding. PMID- 11423557 TI - Kinetic mechanism and regulation of myosin VI. AB - Myosin VI is the only pointed end-directed myosin identified and is likely regulated by heavy chain phosphorylation (HCP) at the actin-binding site in vivo. We undertook a detailed kinetic analysis of the actomyosin VI ATPase cycle to determine whether there are unique adaptations to support reverse directionality and to determine the molecular basis of regulation by HCP. ADP release is the rate-limiting step in the cycle. ATP binds slowly and with low affinity. At physiological nucleotide concentrations, myosin VI is strongly bound to actin and populates the nucleotide-free (rigor) and ADP-bound states. Therefore, myosin VI is a high duty ratio motor adapted for maintaining tension and has potential to be processive. A mutant mimicking HCP increases the rate of P(i) release, which lowers the K(ATPase) but does not affect ADP release. These measurements are the first to directly measure the steps regulated by HCP for any myosin. Measurements with double-headed myosin VI demonstrate that the heads are not independent, and the native dimer hydrolyzes multiple ATPs per diffusional encounter with an actin filament. We propose an alternating site model for the stepping and processivity of two-headed high duty ratio myosins. PMID- 11423558 TI - BACE2 functions as an alternative alpha-secretase in cells. AB - BACE1 and BACE2 define a new subfamily of membrane-anchored aspartyl proteases. Both endoproteases share similar structural organization including a prodomain, a catalytic domain formed via DTG and DSG active site motifs, a single transmembrane domain, and a short C-terminal tail. BACE1 has been identified as the Alzheimer's beta-secretase, whereas BACE2 was mapped to the Down's critical region of human chromosome 21. Herein we show that purified BACE2 can be autoactivated in vitro. Purified BACE2 cleaves human amyloid precursor protein (APP) sequences at the beta-secretase site, and near the alpha-secretase site, mainly at A beta-Phe(20)--Ala(21) and also at A beta-Phe(19)--Phe(20). Alternatively, in cells BACE2 has a limited effect on the beta-secretase site but efficiently cleaves the sequences near the alpha-secretase site. The in vitro specificity of APP processing by BACE2 is distinct from that observed in cells. BACE2 localizes in the endoplasmic reticulum, Golgi, trans-Golgi network, endosomes, and plasma membrane, and its cellular localization patterns depend on the presence of its transmembrane domain. BACE2 chimeras that increase localization of BACE2 in the trans-Golgi network do not change its APP processing patterns. Thus, BACE2 can be distinguished from BACE1 on the basis of autoprocessing of the prosegment, APP processing specificity, and subcellular localization patterns. PMID- 11423559 TI - Control of heterotypic fibril formation by collagen V is determined by chain stoichiometry. AB - Although the collagen V heterotrimer is known to be involved in the control of fibril assembly, the role of the homotrimer in fibrillar organization has not yet been examined. Here, the production of substantial amounts of recombinant collagen V homotrimer has allowed a detailed study of its role in homotypic and heterotypic fibril formation. After removal of terminal regions by pepsin digestion, both the collagen V heterotrimer and homotrimer formed thin homotypic fibrils, thus showing that diameter limitation is at least in part an intrinsic property of the collagen V triple helix. When mixed with collagen I, however, various complementary approaches indicated that the collagen V heterotrimer and homotrimer exerted different effects in heterotypic fibril formation. Unlike the heterotrimer, which was buried in the fibril interior, the homotrimer was localized as thin filamentous structures at the surface of wide collagen I fibrils and did not regulate fibril assembly. Its localization at the fibril surface suggests that the homotrimer can act as a molecular linker between collagen fibrils or macromolecules in the extracellular matrix or both. Thus, depending on their respective distribution in tissues, the different collagen V isoforms might fulfill specific biological functions. PMID- 11423561 TI - Intrarenal and cellular localization of CLC-K2 protein in the mouse kidney. AB - CLC-K2, a kidney-specific member of the CLC chloride channel family, is thought to play an important role in the transepithelial Cl(-) transport in the kidney. This consensus was first reached shortly after it was demonstrated that the mutations of the human CLCNKB gene resulted in Bartter's syndrome type III. To clarify the pathogenesis, the exact intrarenal and cellular localization of CLC K2 by immunohistochemistry of the Clcnk1-/- mouse kidney were investigated by use of an anti-CLC-K antibody that recognized both CLC-K1 and CLC-K2. CLC-K2 is expressed in the thick ascending limb of Henle's loop and distal tubules, where it is localized to the basolateral membranes. The localization of CLC-K2 to these nephron segments strongly implies that CLC-K2 confers the basolateral chloride conductance in the thick ascending limb of Henle's loop and distal tubules, where Cl(-) is taken up by the bumetanide-sensitive Na-K-2Cl cotransporter or the thiazide-sensitive Na-Cl cotransporter at the apical membranes. CLC-K2 expression was also shown to extend into the connecting tubule in the basolateral membrane. CLC-K2 was found in basolateral membranes of the type A intercalated cells residing along the collecting duct. This localization strongly suggests that CLC K2 confers the basolateral conductance in the type A intercalated cells where Cl( ) is taken up by the anion exchanger in exchange for HCO(3)(-) at the basolateral membranes. These aspects of CLC-K2 localization suggest that CLC-K2 is important in Cl(-) transport in the distal nephron segments. PMID- 11423562 TI - Loop diuretic infusion increases thiazide-sensitive Na(+)/Cl(-)-cotransporter abundance: role of aldosterone. AB - Chronic infusion of loop diuretics into animals induces structural and functional changes in the distal nephron. These changes include increases in the activity of the thiazide-sensitive Na(+)/Cl(-)-cotransporter (NCC). The NCC was recently demonstrated to be an aldosterone-induced protein. These experiments were designed to test the hypotheses that chronic loop diuretic infusion, with replacement of NaCl losses, increases NCC protein abundance and that this effect results, in part, from stimulation by aldosterone. Sprague-Dawley rats received vehicle (group 1), furosemide (22 mg/100 g body wt per d) (group 2), or furosemide plus spironolactone (22 and 20 mg/100 g body wt per d, respectively) (group 3). Urine output was higher for groups 2 and 3 than for group 1 (151 +/- 32, 149 +/- 24, and 12 +/- 4 ml, respectively; P < 0.0001). Immunoblot analysis of NCC protein demonstrated that loop diuretics increased NCC protein abundance by nearly 100% (from 2562 +/- 30 to 5248 +/- 151 arbitrary units, P < 0.01). Spironolactone decreased NCC protein abundance by 66% (to 3532 +/- 113 units), compared with the furosemide-treated group (P < 0.005). Northern blot analysis of NCC mRNA demonstrated no significant effect of furosemide (NCC/glyceraldehyde-3 phosphate dehydrogenase ratios: group 1, 0.6 +/- 0.12; group 2, 0.5 +/- 0.05; P > 0.05, NS) These results indicate that increased NCC activity during chronic loop diuretic infusion is associated with increases in NCC protein abundance. A portion of the furosemide effect can be prevented by blockade of mineralocorticoid receptors. PMID- 11423563 TI - Calcitriol controls the epithelial calcium channel in kidney. AB - The recently cloned epithelial Ca2+ channel (ECaC), which is expressed primarily in 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3))-responsive Ca2+ -transporting epithelia, is postulated to constitute the rate-limiting step in active Ca2+ reabsorption. In the present study, the effect of 1,25(OH)(2)D(3) was investigated on ECaC mRNA and protein levels in kidneys of rats that were raised on a vitamin D-depleting diet. This diet decreased the serum 1,25(OH)(2)D(3) concentration significantly, which was accompanied by a marked drop in serum Ca2+ level. Both 1,25(OH)(2)D(3) and Ca2+ levels were normalized within 48 h after 1,25(OH)(2)D(3) administration. In 1,25(OH)(2)D(3)-deficient rats, ECaC mRNA and protein levels of the kidney cortex were significantly decreased compared with the repleted animals, suggesting that 1,25(OH)(2)D(3) exerts its stimulatory effect on Ca2+ reabsorption via increased ECaC expression. In agreement with this observation, the elucidated human ECaC promoter contains several consensus vitamin D-responsive elements. ECaC was restricted to the apical membrane of the distal part of the distal convoluted and the connecting tubule. This conclusion was based on only minor overlap with the localization of the thiazide-sensitive NaCl co-transporter and complete co-localization with the 1,25(OH)(2)D(3) dependent Ca2+ binding protein, calbindin-D(28K). In conclusion, ECaC, present in the distal part of the nephron, is an important target for 1,25(OH)(2)D(3) mediated Ca2+ reabsorption. PMID- 11423564 TI - Use of positron emission tomography to study AT1 receptor regulation in vivo. AB - Increased sodium intake and enhanced sodium sensitivity are implicated in the pathogenesis of hypertension and in the control of a major regulator of BP, the type 1 angiotensin receptor (AT(1) receptor). An in vivo technique to study changes of renal AT(1) receptors by dietary sodium was developed that uses positron emission tomography (PET). PET revealed that renal cortical AT(1) receptor binding was increased in sodium-loaded compared with sodium-deprived dogs, which correlated with ex vivo estimations of AT(1) receptor numbers. Plasma renin activity, angiotensin II, and aldosterone were inversely related to changes in AT(1) receptor binding. These results demonstrate, for the first time in vivo, that the renal AT(1) receptor is inversely related to the activity of the renin angiotensin system, which may provide a compensatory mechanism to prevent inappropriate fluctuations in arterial BP. The ability to measure AT(1) receptor binding in vivo has potential significance for clinical studies of AT(1) receptors, because PET is a noninvasive imaging technique that is readily applicable in humans. PMID- 11423565 TI - Endothelin-1 induces cyclooxygenase-2 expression via nuclear factor of activated T-cell transcription factor in glomerular mesangial cells. AB - Nuclear factor of activated T cells (NFAT) originally was identified as a T-cell specific transcription factor whose activity is regulated by calcineurin, one of the serine-threonine phosphatases. Recent studies have shown that NFAT also is expressed in nonlymphoid cells and plays an important role in various cell functions. It is widely known that treatment with cyclosporin A (CsA), which can inhibit calcineurin/NFAT signaling, results in glomerular dysfunction characterized by a decrease of GFR or glomerulosclerosis, suggesting that NFAT might regulate the glomerular function. However, the precise function of NFAT in glomerular cells remains to be clarified. Herein, evidence has been produced that NFAT2/NFATc, one of five known NFAT isoforms, is expressed in glomerular mesangial cells. Stimulation of mesangial cells with endothelin-1 caused translocation of NFAT2 into the nucleus with a concomitant increase in NFAT2 DNA binding activity, both of which were inhibited by CsA. Furthermore, CsA inhibited endothelin-1-induced cyclooxygenase-2 (COX-2) expression in mesangial cells. NFAT2 bound directly to the GGAAA sequence, which is the minimal consensus sequence for NFAT binding, in a promoter region of rat COX-2 gene, and it enhanced the reporter activity of rat COX-2 promoter in mesangial cells. These findings provide the first evidence that NFAT2 is expressed and regulates COX-2 gene expression in mesangial cells. These results will contribute to evaluation of the precise roles of NFAT in glomerular functions and the CsA-induced nephrotoxicity. PMID- 11423566 TI - Chemokine expression precedes inflammatory cell infiltration and chemokine receptor and cytokine expression during the initiation of murine lupus nephritis. AB - Lupus nephritis is characterized by immune complex deposition and inflammatory cell infiltration. Therefore, the initiation and progression of lupus nephritis in MRL/MpJ Fas(lpr/lpr) (MRL/lpr) mice were investigated, with a focus on the expression of several chemokines and chemokine receptors. Mice were monitored for proteinuria from 6 to 20 wk of age, and kidneys were examined every 2 wk by light microscopy, electron microscopy, and immunohistologic analyses. Furthermore, the expression of chemokines, chemokine receptors, and proinflammatory cytokines was analyzed in ribonuclease protection assays. MRL/lpr mice demonstrated increased expression of monocyte chemoattractant protein-1, regulated upon activation, normal T cell-expressed and -secreted protein, inducible protein of 10 kD, and macrophage inflammatory protein-1beta at week 8. At that time point, levels of circulating and glomerular immune complexes were increased, and no proteinuria or histopathologic signs of renal damage could be observed. As assessed in immunohistochemical and in situ hybridization analyses, monocyte chemoattractant protein-1 and regulated upon activation, normal T cell-expressed and -secreted protein expression was preferentially located in the glomeruli and interstitium. Mononuclear cell infiltration of the kidney was observed by weeks 10 to 12. At week 12, the renal expression of chemokine receptor 1 (CCR1), CCR2, and CCR5 was increased, mice became proteinuric, and renal damage was histologically evident. Finally, the expression of proinflammatory cytokines was detected (weeks 12 to 14). In summary, (1) chemokines are upregulated before inflammatory cell infiltration, proteinuria, and kidney damage are observed; (2) chemokine generation is restricted to sites of subsequent inflammatory cell infiltration, i.e., glomeruli and interstitium; (3) chemokine receptor expression parallels mononuclear cell infiltration; and (4) proinflammatory cytokines are upregulated later, in parallel with inflammatory cell infiltration and the onset of proteinuria. These results support the hypothesis that chemokines initiate leukocyte infiltration and precede proteinuria and renal damage in MRL/lpr mice. PMID- 11423567 TI - Inhibiting the complement system does not reduce injury in renal ischemia reperfusion. AB - The complex pathogenesis of ischemia reperfusion injury (IRI) includes endothelial expression of adhesion molecules, leukocyte recruitment and activation, reactive oxygen species production, and apoptotic and necrotic cell death. A role for complement in IRI of different organs, including kidney, has been proposed on the basis of results of experiments that used pharmacologic inhibitors as well as animals that were deficient in individual complement proteins. Here, renal IRI in mice was examined. Animals that were deficient in C3 had partial protection from IRI induced by 27.5 min of bilateral renal ischemia, followed by 20 h of reperfusion (blood urea nitrogen [BUN] values, 46.6 +/- 6.9 and 68.4 +/- 7.9 mg/dl in C3 -/- and C3 +/+ mice; n = 7 and 8, respectively; P = 0.033). Given the reduction in IRI in C3 -/- mice, it was investigated, by use of the rodent C3 convertase inhibitor CR1-related gene/protein y-Ig (Crry-Ig), whether exogenous administration of a complement inhibitor could lessen renal injury. Despite the use of Crry-Ig in high doses, there was no significant reduction of injury induced by 20 to 30 min of ischemia followed by up to 30 h of reperfusion. Histologic examination revealed acute tubular necrosis and neutrophilic infiltration, both of which correlated significantly with BUN values (P < 0.001). Of interest, C3 deposition around renal tubules was significantly less in animals with IRI, compared with that in unmanipulated controls (P < 0.001). In Crry-Ig-treated animals, C3 deposition was inversely proportional to BUN values (r = -0.63; P < 0.001), which presumably indicates that severe vascular IRI allowed access of the 160 kD Crry-Ig to the interstitium. Thus, renal IRI in mice may have a partial complement dependence, yet pharmacologic inhibition of the complement system does not seem to be effective, likely because of the presence of other mediator systems that operate in parallel. PMID- 11423568 TI - Glomerular injury and tubular loss in adriamycin nephrosis. AB - Glomerular injury manifested by sustained proteinuria usually leads to tubule injury and reduction of the GFR. The current study explored the link between these processes in rats with adriamycin nephrosis. One group of nephrotic rats received a vasopressin V2 receptor blocker (V2X) from 4 to 16 wk after injection of adriamycin, whereas a second group received no treatment (NoRx). V2 receptor blockade increased urine volume without affecting protein excretion. At 16 wk, both groups of nephrotic rats exhibited a marked reduction in GFR in comparison with normal controls (V2X, 0.22 +/- 0.19 ml/min; NoRx, 0.20 +/- 0.11 ml/min; control, 1.23 +/- 0.11 ml/min). Morphologic studies revealed that the majority of glomeruli in nephrotic rats were no longer connected to normal tubule segments (V2X, 81 +/- 21%; NoRx, 85 +/- 18%; control, 1 +/- 2%). Glomeruli without tubules were not, however, globally sclerosed. Disruption of the glomerular tubular junction was associated with the presence of amorphous material separating damaged tubule cells from the basement membrane. Serial sections revealed that this material spread from extensive areas of adhesion between the glomerular tuft and capsule to invest the tubular neck. Reduction of the GFR was strongly correlated with the fraction of glomeruli not connected to normal tubules (r(2) = 0.82; P < 0.0001). V2 receptor blockade did not preserve renal function or structure. These findings suggest that local extension of glomerular injury to destroy the tubule neck is an important cause of loss of renal function in adriamycin nephrosis. PMID- 11423569 TI - The potential of bone marrow-derived cells to differentiate to glomerular mesangial cells. AB - Bone marrow stem cells (BMC) develop into hematopoietic and mesenchymal lineages but have not been known to differentiate into glomerular cells. To investigate whether such differentiation is possible, a search was made for donor glomerular cells in lethally irradiated C57BL/6j (B6) mice given transplants of BMC from syngeneic mice transgenic for green fluorescence protein (GFP) ([GFP-->B6] mice). After the recipients of donor BMC manifested GFP-positive cells in their glomeruli, the numbers of such cells increased markedly, in a time-dependent manner, from 2 wk to 24 wk after bone marrow transplantation. Immunohistochemical analyses revealed that most GFP-positive cells in the glomeruli were neither macrophages nor T cells. With the use of a laser-scanning confocal microscope, GFP-positive cells were observed within the mesangium of [GFP-->B6] mice. Furthermore, indirect immunofluorescence assays demonstrated that desmin-positive cells in the glomeruli of these chimeric mice were also positive for GFP. Among glomerular cells isolated from [GFP-->B6] mice 24 wk after bone marrow transplantation and then cultured, the majority of cells (approximately 84%) stained for desmin and approximately 60% of the desmin-positive cells expressed GFP. In addition, these GFP-positive cells in the cultures contracted in response to angiotensin II stimulation. These results suggest that bone marrow-derived cells may have the potential to differentiate into glomerular mesangial cells. PMID- 11423570 TI - Cell proliferation in the loop of henle in the developing rat kidney. AB - In the developing rat kidney, there is no separation of the medulla into an outer and inner zone. At the time of birth, ascending limbs with immature distal tubule epithelium are present throughout the renal medulla, all loops of Henle resemble the short loop of adult animals, and there are no ascending thin limbs. It was demonstrated previously that immature thick ascending limbs in the renal papilla are transformed into ascending thin limbs by apoptotic deletion of cells and transformation of the remaining cells into a thin squamous epithelium. However, it is not known whether this is the only source of ascending thin limb cells or whether cell proliferation occurs in the segment undergoing transformation. This study was designed to address these questions and to identify sites of cell proliferation in the loop of Henle. Rat pups, 1, 3, 5, 7, and 14 d old, received a single injection of 5-bromo-2'-deoxyuridine (BrdU) 18 h before preservation of kidneys for immunohistochemistry. Thick ascending and descending limbs were identified by labeling with antibodies against the serotonin receptor, 5-HT(1A), and aquaporin-1, respectively. Proliferating cells were identified with an antibody against BrdU. BrdU-positive cells in descending and ascending limbs of the loop of Henle were counted and expressed as percentages of the total number of aquaporin-1-positive and 5-HT(1A)-positive cells in the different segments. In the developing kidney, numerous BrdU-positive nuclei were observed in the nephrogenic zone. Outside of this location, BrdU-positive tubule cells were most prevalent in medullary rays in the inner cortex and in the outer medulla. BrdU labeled cells were rare in the papillary portion of the loop of Henle and were not observed in the lower half of the papilla after 3 d of age. BrdU-labeled nuclei were not observed in segments undergoing transformation or in newly formed ascending thin limb epithelium. It was concluded that the growth zone for the loop of Henle is located around the corticomedullary junction, and the ascending thin limb is mainly, if not exclusively, derived from cells of the thick ascending limb. PMID- 11423571 TI - Altered signaling and regulatory mechanisms of apoptosis in focal and segmental glomerulosclerosis. AB - The purpose of this study was to investigate signaling and regulatory mechanisms of apoptosis in a model of focal and segmental glomerulosclerosis. Sprague-Dawley rats received two doses of puromycin aminonucleoside (PAN) (day 0 and week 3) and a uninephrectomy (PAN model). Apoptosis was detected with the use of the terminal deoxynucleotidyl transferase mediated dUTP nick end labeling technique. Bax, Bcl 2, Fas, and Fas ligand expression was analyzed by competitive reverse transcription-PCR. Bax, Bcl-2, and Fas mRNA were localized by in situ hybridization. Renal function was transiently impaired after the first PAN dose. After the second PAN dose, further progressive renal impairment, tubular atrophy, interstitial fibrosis, and glomerulosclerosis were evident. Eighteen percent of PAN samples demonstrated up to 4 apoptotic cells/50 glomeruli, compared with 7% of sham controls (not significant). No consistent significant change in glomerular Bax, Bcl-2, Fas, and Fas ligand mRNA was evident by reverse transcription-PCR, although focal increases in glomerular Bcl-2 mRNA were demonstrated by in situ hybridization. In the tubulointerstitium, apoptosis was increased from weeks 1 to 12 (P < 0.01 PAN versus sham), correlated to renal function and tubulointerstitial injury (P < 0.01). Total renal Bax, Fas, and Fas ligand mRNA were upregulated in the PAN model, peaking at week 17 (P < 0.01 versus sham), whereas Bcl-2 mRNA was not significantly different in PAN versus sham controls. In situ hybridization in the PAN model demonstrated prominent Bax mRNA in dilated tubules and infiltrating leukocytes. Fas mRNA signal was localized to tubular epithelial cells and leukocytes. The results suggest that altered apoptotic signaling and regulatory mechanisms contribute to the tubulointerstitial injury in this model. PMID- 11423572 TI - Impaired angiogenesis in the remnant kidney model: I. Potential role of vascular endothelial growth factor and thrombospondin-1. AB - Few studies have examined the role of the microvasculature in progressive renal disease. It was hypothesized that impaired angiogenesis might occur in the diseased kidney and could contribute to renal scarring. Progressive renal disease was induced in rats by 5/6 renal ablation and those rats were compared with sham operated control animals at multiple time points, for examination of changes in the microvasculature and the expression of angiogenic factors. An early angiogenic response was documented in remnant kidneys, with increases in the proliferation of peritubular (1 wk) and glomerular (2 wk) endothelial cells. Subsequently, however, there was a decrease in endothelial cell proliferation, which was reduced to levels below those of sham-treated animals, in conjunction with interstitial expression of the antiangiogenic factor thrombospondin-1 (TSP 1) and decreased tubular expression of the proangiogenic factor vascular endothelial growth factor (VEGF). Both the increase in TSP-1 expression and the loss of VEGF expression were correlated with capillary loss and the development of glomerulosclerosis and interstitial fibrosis. Progressive macrophage infiltration was correlated both spatially and quantitatively with the sites of absent or diminished VEGF expression. In addition, macrophage-associated cytokines (interleukin-1beta, interleukin-6, and tumor necrosis factor-alpha) inhibited VEGF mRNA expression and protein secretion by cultured tubular epithelial cells of the medullary thick ascending limb, under both normoxic and hypoxic conditions. Impaired angiogenesis characterizes the remnant kidney model and is correlated with progression. The impaired angiogenesis may be mediated by alterations in the renal expression of TSP-1 and VEGF, with the latter being regulated by macrophage-associated cytokines. PMID- 11423573 TI - Impaired angiogenesis in the remnant kidney model: II. Vascular endothelial growth factor administration reduces renal fibrosis and stabilizes renal function. AB - Impaired angiogenesis and decreased vascular endothelial growth factor (VEGF) expression were recently documented in the remnant kidney (RK) model of progressive renal failure. VEGF (50 microg/kg, twice daily) was administered to RK rats between weeks 4 and 8 after surgery, and rats were euthanized at week 8 for histologic study. During the administration of VEGF (n = 7) or vehicle (n = 6), systemic BP was comparable in the two groups. VEGF treatment resulted in improved renal function and lower mortality rates, compared with the vehicle treated group. Renal histologic analyses confirmed a 3.5-fold increase in glomerular endothelial cell proliferation (0.14 +/- 0.03 versus 0.04 +/- 0.02 proliferating endothelial cells/glomerulus, VEGF versus vehicle, P < 0.05), a twofold increase in peritubular capillary endothelial cell proliferation (1.60 +/ 0.30 versus 0.78 +/- 0.17 cells/mm(2), VEGF versus vehicle, P < 0.01), a threefold decrease in peritubular capillary rarefaction (P < 0.01), and a twofold increase in endothelial nitrix oxide synthase expression (P < 0.05) in the VEGF treated group; an eightfold increase in urinary nitrate/nitrite levels (P < 0.05) was also noted. Although the difference in glomerulosclerosis scores did not reach statistical significance (0.67 +/- 0.42 versus 1.22 +/- 0.63, VEGF versus vehicle; range, 0 to 4; P = NS), VEGF-treated rats exhibited less interstitial collagen type III deposition (9.32 +/- 3.26 versus 17.45 +/- 7.50%, VEGF versus vehicle, P < 0.01) and reduced tubular epithelial cell injury, as manifested by osteopontin expression (5.57 +/- 1.60 versus 9.58 +/- 3.45%, VEGF versus vehicle, P < 0.01). In conclusion, VEGF treatment reduces fibrosis and stabilizes renal function in the RK model. The use of angiogenic factors may represent a new approach to the treatment of kidney disease. PMID- 11423574 TI - Response to single and divided doses of Shiga toxin-1 in a primate model of hemolytic uremic syndrome. AB - Postdiarrheal hemolytic uremic syndrome is caused by Shiga toxin (Stx)-producing Escherichia coli. It was shown previously that the baboon, like the human, has glycolipid receptors for Stx in the gut and the kidney and that a single 50- to 200-ng/kg intravenous dose of purified Stx-1 results in thrombocytopenia, hemolytic anemia, and renal thrombotic microangiopathy. For further characterization of factors that modulate disease expression, the baboon's response to the intravenous administration of 100 ng/kg Stx-1 given either rapidly as a single bolus or slowly as four 25-ng/kg doses at 12-h intervals was compared. Animals that received the Stx-1 as a single dose developed thrombocytopenia, schistocytosis, and acute renal failure. Urinary but not plasma tumor necrosis factor-alpha concentrations rose significantly by 6 h and then declined rapidly. Urinary and plasma interleukin-6 concentrations rose later. Glomeruli showed reduced patency of capillary loops, fragmented red blood cells, fibrin and platelet microthrombi, necrosis and detachment of endothelial cells, and accumulation of flocculent material in subendothelial spaces. Damage to tubular epithelium and peritubular capillary endothelium also was seen. Animals that received four divided doses of Stx-1 developed no clinical or histologic features of hemolytic uremic syndrome. It is concluded that in the primate model, disease expression is modulated by the rate of Stx administration, and it is speculated that in the human, the rate of Stx absorption from the gut is one determinant of disease severity. PMID- 11423575 TI - Relative potencies of 1,25-(OH)(2)D(3) and 19-Nor-1,25-(OH)(2)D(2) on inducing differentiation and markers of bone formation in MG-63 cells. AB - 19-Nor-1,25-(OH)(2)D(2), an analog of 1,25-(OH)(2)D(3), is used to treat secondary hyperparathyroidism because it suppresses parathyroid hormone synthesis and secretion with lower calcemic and phosphatemic activities. 19-Nor-1,25 (OH)(2)D(2) is approximately 10 times less active than 1,25-(OH)(2)D(3) in promoting bone resorption, which accounts in part for the low potency of this analog in increasing serum calcium and phosphorus. Concern that 19-nor-1,25 (OH)(2)D(2) also could be less potent than 1,25-(OH)(2)D(3) on bone formation led to a comparison of the potency of both compounds on osteoblasts. In the human osteoblast-like cell line MG-63, 1,25-(OH)(2)D(3) and 19-nor-1,25-(OH)(2)D(2) had a similar potency in upregulating vitamin D receptor content and suppressing proliferation. Both sterols caused a similar reduction in DNA content and proliferating cell nuclear antigen protein expression. Time-course and dose response studies on 1,25-(OH)(2)D(3) and 19-nor-1,25-(OH)(2)D(2) induction of the marker of bone formation, osteocalcin, showed overlapping curves. The effects on alkaline phosphatase (ALP) activity also were studied in MG-63 cells that had been co-treated with either sterol and transforming growth factor-beta, an enhancer of 1,25-(OH)(2)D(3)-induced ALP activity in this cell line. Transforming growth factor-beta alone had no effect, whereas 1,25-(OH)(2)D(3) and 19-nor-1,25 (OH)(2)D(2) increased ALP activity similarly. These studies demonstrate that 19 nor-1,25-(OH)(2)D(2) has the same potency as 1,25-(OH)(2)D(3) not only in inducing vitamin D receptor content, osteocalcin levels, and ALP activity but also in controlling osteoblastic growth. Therefore, it is unlikely that 19-nor 1,25-(OH)(2)D(2) would have deleterious effects on bone remodeling. PMID- 11423576 TI - Long-term effects of arterial stenting on kidney function for patients with ostial atherosclerotic renal artery stenosis and renal insufficiency. AB - It is uncertain whether renal artery stent placement in patients with atherosclerotic renovascular renal failure can prevent further deterioration of renal function. Therefore, the effects of renal artery stent placement, followed by patency surveillance, were prospectively studied in 63 patients with ostial atherosclerotic renal artery stenosis and renal dysfunction (i.e., serum creatinine concentrations of >120 micromol/L (median serum creatinine concentration, 171 micromol/L; serum creatinine concentration range, 121 to 650 micromol/L). Pre-stent renal (dys) function was stable for 28 patients and declining for 35 patients (defined as a serum creatinine concentration increase of > or =20% in 12 mo). The median follow-up period was 23 mo (interquartile range, 13 to 29 mo). Angioplasty to treat restenosis was performed in 12 cases. Five patients reached end-stage renal failure within 6 mo, and this was related to stent placement in two cases. Two other patients died or were lost to follow up monitoring within 6 mo, with stable renal function. For the remaining 56 patients, the treatment had no effect on serum creatinine levels if function had previously been stable; if function had been declining, median serum creatinine concentrations improved in the first 1 yr [from 182 micromol/L (135 to 270 micromol/L ) to 154 micromol/L (127 to 225 micromol/L ); P < 0.05] and remained stable during further follow-up monitoring. In conclusion, stent placement, followed by patency surveillance, to treat ostial atherosclerotic renal artery stenosis can stabilize declining renal function. For patients with stable renal dysfunction, the usefulness is less clear. The possible advantages must be weighed against the risk of renal failure advancement with stent placement. PMID- 11423577 TI - Renal monoclonal immunoglobulin deposition disease: the disease spectrum. AB - This study reports the clinicopathologic findings and outcome in 34 patients with renal monoclonal immunoglobulin deposition disease (MIDD), which included 23 light-chain DD (LCDD), 5 light- and heavy-chain DD (LHCDD), and 6 heavy-chain DD (HCDD). A total of 23 patients had pure MIDD, whereas 11 patients had LCDD with coexistent myeloma cast nephropathy (LCDD & MCN). Renal biopsy diagnosis preceded clinical evidence of dysproteinemia in 68% of all cases. By immunofluorescence, the composition of deposits included 11kappa/1lambda (LCDD), 3IgGkappa/2IgGlambda (LHCDD), 5gamma/1alpha (HCDD), and 10kappa/1lambda (LCDD & MCN). Patients with pure MIDD presented with mean serum creatinine of 4.2 mg/dl, nephrotic proteinuria, and hypertension. Cases of HCDD were associated with a CH1 deletion and frequently had hypocomplementemia and a positive hepatitis C virus antibody but negative hepatitis C virus PCR. LCDD & MCN is a morphologically and clinically distinct entity from pure MIDD, presenting with higher creatinine (mean, 7.8 mg/dl; P = 0.01), greater dialysis dependence (64 versus 26%; P = 0.053), subnephrotic proteinuria, and less nodular glomerulopathy (18 versus 100%; P < 0.0001). Multiple myeloma was more frequently diagnosed in LCDD & MCN than in pure MIDD (91 versus 31%; P = 0.025). Renal and patient survivals were significantly worse in patients with LCDD & MCN (mean, 4 and 22 mo, respectively), compared with patients with pure MIDD (mean, 22 and 54 mo). Chemotherapy stabilized or improved renal function in 10 of 15 patients (67%) with pure MIDD who presented with creatinine of <5.0 mg/dl, emphasizing the importance of early detection. On multivariate analysis, initial creatinine was the only predictor of renal and patient survival in pure MIDD, underscoring the prognostic significance of the renal involvement. PMID- 11423578 TI - Antineutrophil cytoplasmic autoantibody-associated glomerulonephritis in children. AB - Aretrospective investigation was conducted by members of the Japanese Society for Pediatric Nephrology from 1990 to 1997 to define the clinical features and outcome of antineutrophil cytoplasmic autoantibody (ANCA)-associated glomerulonephritis in children. Thirty-four ANCA-seropositive Japanese pediatric patients with biopsy-proven pauci-immune necrotizing crescentic glomerulonephritis were identified. Of these, 3 cases associated with Wegener's granulomatosis were excluded because of the small sample size. Among the 31 patients studied, 10 had a diagnosis of necrotizing crescentic glomerulonephritis alone and 21 had microscopic polyangiitis. Females predominated (87%), and the median age at onset was 12 yr. Twenty-six patients received treatment with cyclophosphamide and corticosteroids, and five patients received treatment with corticosteroids alone; 84% of patients achieved remission, and 39% of responders relapsed in a median of 24 mo. ANCA titers correlated with response to treatment and disease activity, with some exceptions. Patients were followed for a median of 42 mo (range, 3 to 96 mo). Nine of 31 patients (29.0%) progressed to end-stage renal disease, 6 (19.4%) had reduced renal function, and 15 (48.4%) had normal renal function at the last observation. One patient (3.2%) died from cytomegalovirus infection 3 mo after initiation of therapy. Life-table analysis showed 75% renal survival at 39 mo. Patients who subsequently developed end-stage renal disease (n = 9) had significantly higher average peak serum creatinine levels and more chronic pathologic lesions at diagnosis compared with patients with favorable renal outcome (n = 15). In conclusion, our clinical experience suggests that the clinical disease spectrum of ANCA-associated glomerulonephritis is similar in pediatric and adult patients, but there is a female predominance in children. PMID- 11423579 TI - Management of patients with chronic renal insufficiency in the Northeastern United States. AB - Comorbid conditions that develop during chronic renal insufficiency (CRI) contribute to the high morbidity and mortality among patients with end-stage renal disease (ESRD). Thus, appropriate management during CRI may lead to improved ESRD outcomes. A retrospective cohort study was performed to describe the management of patients with CRI. A total of 602 patients with CRI (creatinine > or =1.5 mg/dl for women and > or =2.0 mg/dl for men) were seen between October 1994 and September 1998 at five nephrology outpatient clinics in the Boston area. The mean (SD) age of the patients was 63 (15.5) yr, and 53% were male. At the first nephrology visit, mean (SD) serum creatinine was 3.2 (1.6) mg/dl, and mean (SD) predicted GFR was 22.3 (8.9) ml/min per 1.73 m(2). Laboratory tests for iron levels were performed in only 18% of patients, serum parathyroid hormone levels were obtained in only 15%, lipid studies were obtained in fewer than half, and among patients with diabetes, only 28% had a glycosylated hemoglobin level measured. A hematocrit <30% was present in 38%, and abnormal calcium-phosphorus metabolism was noted in 55%. Only 59% of patients who had hematocrit <30% received recombinant human erythropoietin. Among patients who received recombinant human erythropoietin, only 47% received iron. Angiotensin-converting enzyme inhibitor use was recorded for only 65% of patients with diabetes (49% of patients overall). Among patients who were known to have progressed to ESRD, only 41% had permanent access placed before initiation of dialysis. There seems to be room for improvement in the management of patients with CRI, which could result in a slower rate of progression of CRI and reduced severity of comorbid conditions. PMID- 11423580 TI - Cardiac natriuretic peptides are related to left ventricular mass and function and predict mortality in dialysis patients. AB - This study was designed to investigate the relationship among brain natriuretic peptide (BNP) and atrial natriuretic peptide (ANP) and left ventricular mass (LVM), ejection fraction, and LV geometry in a large cohort of dialysis patients without heart failure (n = 246) and to test the prediction power of these peptides for total and cardiovascular mortality. In separate multivariate models of LVM, BNP and ANP were the strongest independent correlates of the LVM index. In these models, the predictive power of BNP was slightly stronger than that of ANP. Both natriuretic peptides also were the strongest independent predictors of ejection fraction, and again BNP was a slightly better predictor of ejection fraction than ANP. In separate multivariate Cox models, the relative risk of death was significantly higher in patients of the third tertile of the distribution of BNP and ANP than in those of the first tertile (BNP, 7.14 [95% confidence interval (CI), 2.83 to 18.01, P = 0.00001]; ANP, 4.22 [95% CI, 1.79 to 9.92, P = 0.001]), and a similar difference was found for cardiovascular death (BNP, 6.72 [95% CI, 2.44 to 18.54, P = 0.0002]; ANP, 3.80 [95% CI, 1.44 to 10.03, P = 0.007]). BNP but not ANP remained as an independent predictor of death in a Cox's model including LVM and ejection fraction. Cardiac natriuretic peptides are linked independently to LVM and function in dialysis patients and predict overall and cardiovascular mortality. The measurement of the plasma concentration of BNP and ANP may be useful for risk stratification in these patients. PMID- 11423581 TI - Prevalence and clinical correlates of coronary artery disease among new dialysis patients in the United States: a cross-sectional study. AB - Despite the high prevalence of coronary artery disease (CAD) among patients with end-stage renal disease (ESRD), few studies have identified clinical correlates using national data. The purpose of this study was to determine the prevalence and clinical associations of CAD in a national random sample of new ESRD in the United States in 1996/1997 (n = 4025). Data on demographic characteristics and comorbidities were obtained from the Dialysis Morbidity and Mortality Study, Wave 2. The principal outcome was CAD, defined as the presence of a previous history of CAD, myocardial infarction, or angina, coronary artery bypass surgery, coronary angioplasty, or abnormal coronary angiographic findings. Multivariate logistic regression analysis was used to assess the relationship of conventional factors and proposed uremic factors to the presence of CAD. CAD was present in 38% of patients. Of the total cohort, 17% had a history of myocardial infarction and 23% had angina. Several conventional risk factors, including advancing age, male gender, diabetes mellitus, and smoking, were significantly associated with CAD. Of the proposed uremic factors, lower serum albumin levels but higher residual renal function and higher hematocrit values were significantly associated with the presence of CAD. Vascular comorbid conditions, structural cardiac abnormalities, white race, and geographic location were also strongly correlated with the presence of CAD. This national study suggests that several conventional CAD risk factors may also be risk factors for CAD among the ESRD population. This study identifies nonconventional factors such as serum albumin levels, vascular comorbid conditions, and structural cardiac abnormalities as important disease correlates. Future logitudinal studies are required to explore the relative importance of the relationships observed here. PMID- 11423582 TI - Dynamic changes of the total pore area available for peritoneal exchange in children. AB - The most important of the parameters that describe exchange across the peritoneal membrane is the total pore area over diffusion distance (A(0)/Deltax). It determines the rate of diffusion and mainly seems to reflect the number of capillaries available for exchange. In the present study, a simplified three-pore analysis was used to estimate A(0)/Deltax from peritoneal equilibration tests. Two groups of children (mean age, 9.5 yr) who were on chronic peritoneal dialysis underwent studies with peritoneal equilibration tests. In the first group of children, three levels of fill volumes were used in each patient. In the second group of patients, the effects of posture and dwell time were analyzed from four consecutive peritoneal dialysis samples obtained after 15, 30, 60, and 90 min. As the fill volume was raised from 800 to 1400 ml/m(2) BSA, the steady-state A(0)/Deltax increased significantly by 21%, i.e., from 19,900 +/- 1200 to 24,000 +/- 1450 cm(2)/cm per 1.73 m(2) (n = 8). A further increase to 2000 ml/m(2) did not result in any change of A(0)/Deltax. Moreover, steady-state A(0)/Deltax fell significantly when the patients were standing, 21,900 +/- 1400 compared with 29,400 +/- 1330 cm(2)/cm per 1.73m(2) (n = 6) obtained in the supine position. There was a transient (<30 min) increase in A(0)/Deltax initially during the dwell, probably reflecting vasodilation and recruitment of capillaries. It is concluded that factors such as the intraperitoneal fill volume, posture, and dwell time all dynamically affect the total pore area available for exchange. PMID- 11423583 TI - Effect of ibandronate on bone loss and renal function after kidney transplantation. AB - Severe osteoporosis frequently is observed after organ transplantation. In kidney transplantation, it adds to pre-existing renal bone disease and strategies to prevent osteoporosis are not established. Eighty kidney recipients were included in a randomized controlled prospective intervention trial. Treated patients (n = 40) received an injection of ibandronate, a bisphosphonate, immediately before and at 3, 6, and 9 mo after transplantation. The primary outcome measured was the change in bone mineral density. Secondary measures included graft outcome, spinal deformities, fracture rate, body height, and hormonal and metabolic data. Loss of spongy and cortical bone after transplantation was prevented by ibandronate. Changes of bone mineral density (ibandronate versus controls) were as follows: lumbar spine, -0.9 +/- 6.1% versus -6.5 +/- 5.4% (P < 0.0001); femoral neck, +0.5 +/- 5.2% versus -7.7 +/- 6.5% (P < 0.0001); and midfemoral shaft, +2.7 +/- 12.2% versus -4.0 +/- 10.9% (P = 0.024). Fewer spinal deformities developed with ibandronate (7 patients with 7 deformities versus 12 patients with 23 deformities; P = 0.047). Loss of body height was 0.5 +/- 1.0 cm versus 1.1 +/- 1.0 cm in control subjects (P = 0.040). Two bone fractures occurred in each group. There were fewer acute rejection episodes with ibandronate (11 versus 22; P = 0.009). Graft function after 1 yr was comparable. Bone loss, spinal deformation, and loss of body height during the first year after kidney transplantation are prevented by injection of ibandronate at intervals of 3 mo. The smaller number of rejection episodes of the ibandronate-treated group should be confirmed and its mechanism should be explored in additional studies. PMID- 11423584 TI - Increased immunogenicity and cause of graft loss of old donor kidneys. AB - Donor age was identified recently as a major factor that determines long-term outcome after transplantation, but the mechanism that is responsible for increased graft loss of old donor kidneys is unknown. The influence of donor age on graft survival was assessed retrospectively in 514 consecutive first cadaveric transplants that were treated with cyclosporine maintenance immunosuppression. Donor age > or =50 yr (relative risk [RR] = 1.7; 95% confidence interval [CI], 1.2 to 2.6), acute rejection (RR = 2.0; 95% CI, 1.3 to 3.0), and type of rejection (RR = 3.3; 95% CI, 2.0 to 5.3) had a significant impact on graft survival. However, when subsets of patients who entered subsequent intervals after transplantation were analyzed, donor age was not an independent predictive factor of graft loss. Donor age (RR = 1.53; 95% CI, 1.19 to 1.98), human leukocyte antigen-DR mismatch (RR = 2.28; 95% CI, 1.78 to 2.92), and recipient age (RR = 1.34; 95% CI, 1.05 to 1.72) were associated significantly with acute rejection episodes. Delayed graft function alone was not associated independently with the occurrence of early acute rejection (RR = 1.24; 95% CI, 0.96 to 1.61). The timing of the rejection episodes of old donor kidneys was not different, and the excess rejection prevalence was attributable entirely to interstitial (grade I) types of rejection. Interstitial rejection episodes in kidneys from old donors had a significant (P < 0.05) negative impact on graft survival. Beyond the first year, poor renal function and proteinuria were significant risk factors for graft loss, regardless of rejection. Our data fit best the hypothesis that increased graft loss of older donor kidneys results from an increased incidence of acute interstitial rejection episodes in the early posttransplantation months. It is proposed that kidneys from older donors are more immunogenic than kidneys from young donors and that acute rejection episodes result in functional deterioration. Contrary to interstitial rejection in kidneys from younger donors, kidneys from old donors seem to have an impaired ability to restore tissue. PMID- 11423586 TI - The microinflammatory state in uremia: causes and potential consequences. AB - Mortality is markedly elevated in patients with end-stage renal disease. The leading cause of death is cardiovascular disease. Lipoprotein levels are only slightly elevated in dialysis patients, and cardiovascular risk is inversely correlated with serum cholesterol, suggesting that a process other than hyperlipidemia plays a role in the incidence of cardiovascular disease. Hypoalbuminemia, ascribed to malnutrition, has been one of the most powerful risk factors that predict all-cause and cardiovascular mortality in dialysis patients. The presence of inflammation, as evidenced by increased levels of specific cytokines (interleukin-6 and tumor necrosis factor alpha) or acute-phase proteins (C-reactive protein and serum amyloid A), however, has been found to be associated with vascular disease in the general population as well as in dialysis patients. The process of inflammation, also called the acute-phase response, additionally causes loss of muscle mass and changes in plasma composition decreases in serum albumin, prealbumin, and transferrin levels, also associated with malnutrition. Inflammation alters lipoprotein structure and function as well as endothelial structure and function to favor atherogenesis and increases the concentration of atherogenic proteins in serum, such as fibrinogen and lipoprotein (a). Inflammation in dialysis patients is episodic. The causes are likely to be multifactorial and include vascular access infection, less-than sterile dialysate, dialysate back leak, and nonbiocompatible membranes in addition to clinically apparent infection. In addition, proinflammatory compounds, such as advanced glycation end products, accumulate in renal failure, and defense mechanisms against oxidative injury are reduced, contributing to inflammation and to its effect on the vascular endothelium. PMID- 11423587 TI - Light chain deposition disease: a model of glomerulosclerosis defined at the molecular level. PMID- 11423588 TI - The multiplication principle as the basis for concentrating urine in the kidney. AB - Originally published as "Das Multipikationsprinzip als Grundlage der Harnkonzentrierung in der Niere", in Zeitschrift fur Elektrochemie und angewandte physikalische Chemie 55(6):539-558, 1951. PMID- 11423589 TI - Medical education and practice in the information age. PMID- 11423590 TI - Phyllodes tumours. AB - Phyllodes tumours are rare fibroepithelial lesions that account for less than 1% of all breast neoplasms. With the non-operative management of fibroadenomas widely adopted, the importance of phyllodes tumours today lies in the need to differentiate them from other benign breast lesions. All breast lumps should be triple assessed and the diagnosis of a phyllodes tumour considered in women, particularly over the age of 35 years, who present with a rapidly growing "benign" breast lump. Treatment can be by either wide excision or mastectomy provided histologically clear specimen margins are ensured. Nodal metastases are rare and routine axillary dissection is not recommended. Few reliable clinical and histological prognostic factors have been identified. Local recurrence occurs in approximately 15% of patients and is more common after incomplete excision. It can usually be controlled by further surgery. Repeated local recurrence has been reported without the development of distant metastases or reduced survival. Approximately 20% of patients with malignant phyllodes tumours develop distant metastases. Long term survival with distant metastases is rare. The role of chemotherapy, radiotherapy, and hormonal manipulation in both the adjuvant and palliative settings remain to be defined. PMID- 11423591 TI - Infliximab treatment for Crohn's disease. PMID- 11423592 TI - Therapeutic value of octreotide for patients with severe dumping syndrome--a review of randomised controlled trials. PMID- 11423593 TI - Ethical, professional, and legal obligations in clinical practice: a series of discussion topics for postgraduate medical education. Topic 4: Confidentiality. PMID- 11423594 TI - Is transcutaneous electrical nerve stimulation an effective analgesia during colonoscopy? AB - OBJECTIVES: To evaluate the efficacy of transcutaneous electrical nerve stimulation (TENS) as analgesia during colonoscopy. DESIGN: In a randomised controlled trial, patients undergoing diagnostic colonoscopy were assigned to one of three groups: standard medication only (midazolam); active TENS plus standard medication; or non-functioning TENS and standard medication. Efficacy of TENS was determined using numerical rating scores for pain and the post-procedural evaluation questionnaire. SETTING: Patients undergoing diagnostic colonoscopy in a teaching hospital. MAIN OUTCOME: There was no statistically significant differences between the three groups. However in the active TENS group there was a greater variation in "physical discomfort" and "psychological distress", suggesting TENS may be effective in subgroup of patients. PMID- 11423595 TI - Quadruple therapy for symptomatic spontaneous duodenal ulcer disease. AB - AIM: To investigate Helicobacter pylori eradication in duodenal ulcer patients with a new regimen, lansoprazole 30 mg daily for one or four weeks plus twice daily tetracycline 500 mg, clarithromycin 250 mg, and metronidazole 400 mg. BACKGROUND: Spontaneous duodenal ulcer is regularly associated with H pylori, and permanent cure follows eradication of this bacterium. Numerous treatments have been proposed and none is ideal, possibly because of primary or acquired antibiotic resistance. Quadruple regimens with proton pump inhibitor therapy and three antibiotics offer promise as the most effective therapy. METHODS: From November 1995 all patients with spontaneous duodenal ulcer were offered quadruple therapy. A month after completion a carbon 14 urea breath test (UBT) was performed. Sensitivity of H pylori to the antibiotics used was tested in 1992-3, 1996, and 1999. RESULTS: A total of 331 patients were treated; 313 attended for a UBT, of which 299 were negative (95.5%). Of those patients who had an endoscopy with positive urease test immediately before treatment, 95/101 (94.0%) on lansoprazole for one week and 116/121 (95.8%) on lansoprazole for four weeks had a negative UBT. H pylori antibiotic sensitivity did not change. CONCLUSION: This regimen produced some of the best results yet seen and may be generally recommended as first line therapy. PMID- 11423596 TI - Electrophysiological course of uraemic neuropathy in haemodialysis patients. AB - The objective of this study was to confirm electrophysiologically both the presence and course of uraemic neuropathy in haemodialysis patients. Nerve conduction studies of the lower extremities were done in 70 haemodialysis patients and 20 normal volunteers. Compared with that in normal volunteers, the distal motor latency in the tibial nerve of patients was prolonged significantly (p<0.05), and the minimal F wave latency in the tibial nerve was also prolonged significantly (p<0.05). Motor conduction velocity in the tibial nerve was reduced significantly (p<0.05), and sensory nerve conduction velocity in the medial plantar nerve also was reduced significantly (p<0.05). These results suggest the presence of uraemic neuropathy in haemodialysis patients. Twenty patients were investigated by a follow up study five years later. Parameters from F wave conduction studies, which were thought to be the most useful in the evaluation of neuropathy, showed no significant differences between the initial and follow up trials. These observations suggest that uraemic neuropathy does not progress during haemodialysis. These results also suggest that most haemodialysis patients showed electrophysiological evidence of uraemic neuropathy, but no remarkable electrophysiological change in uraemic neuropathy during haemodialysis was recognised. PMID- 11423597 TI - Do computer generated ECG reports improve interpretation by accident and emergency senior house officers? AB - OBJECTIVES: To determine whether access to a computer generated electrocardiogram (ECG) report can reduce errors of interpretation by senior house officers (SHOs) in an accident and emergency department. METHODS: Ten SHOs were asked to interpret 50 ECGs each: 25 with computer generated reports, 25 without. Their answers, and the computer generated reports, were compared with a "gold standard" produced by two experienced clinicians. The primary outcome measure was the proportion of major errors of interpretation. RESULTS: The computer reading system made two major errors (4%, 95% confidence interval (CI) 1.1% to 13.5%) compared with the gold standard. Access to the computer report did not significantly reduce major errors among SHOs (46 (18.4%) with report v 56 (22.4%) without, odds ratio 0.64, 95% CI 0.36% to 1.14%, p=0.13) or improve the proportion completely correct (104 (41.6%) with report v 91 (36.4%) without, odds ratio 1.43, 95% CI 0.88 to 2.33, p=0.15). CONCLUSIONS: SHOs have a high error rate when interpreting ECGs, which is not significantly reduced by access to a computer generated report. Junior doctors should continue to seek expert senior help when they have to interpret a difficult ECG. PMID- 11423598 TI - Intraoperative glove perforation--single versus double gloving in protection against skin contamination. AB - Surgeons have the highest risk of contact with patients' blood and body fluids, and breaches in gloving material may expose operating room staff to risk of infections. This prospective randomised study was done to assess the effectiveness of the practice of double gloving compared with single gloving in decreasing finger contamination during surgery. In 66 consecutive surgical procedures studied, preoperative skin abrasions were detected on the hands of 17.4% of the surgeons. In the double gloving pattern, 32 glove perforations were observed, of which 22 were in the outer glove and 10 in the inner glove. Only four outer glove perforations had matching inner glove perforations, thus indicating that in 82% of cases when the outer glove is perforated the inner glove will protect the surgeon's hand from contamination. The presence of visible skin contamination was also higher in perforation with the single gloving pattern (42.1%) than with the double gloving pattern (22.7%). An overwhelming majority of glove perforations (83.3%) went unnoticed. Double gloving was accepted by the majority of surgeons, especially with repeated use. It is recommended that double gloves are used routinely in all surgical procedures in view of the significantly higher protection it provides. PMID- 11423599 TI - Trends in the smoking habits of young adults with diabetes. AB - OBJECTIVES: To examine changes in the prevalence of smoking in young adult diabetic patients between 1990 and 1999. SETTING: Walton Diabetes Centre, University Hospital Aintree, Liverpool, UK. DESIGN: Direct questioning as well as the urinary cotinine:creatinine ratio were used to assess the smoking habits of 99 young type 1 diabetic patients in 1991 (mean age 21.5 years, duration of diabetes 7.3 years), and in 112 similar patients in 1999 (mean age 23.4 years, duration of diabetes 9.6 years). RESULTS: The admitted smoking rate was 31/99 (31%) in 1990 compared with 31/112 (28%) in 1999 (not significant). However, in 1990 there were an additional 17 "covert" smokers (patients who denied smoking, but had an unequivocally raised urinary cotinine:creatinine ratio), but only three in 1999 (p<0.05). This gave a corrected validated smoking rate of 48/99 (48%) in 1990 and 34/112 (30%) in 1999, representing a significant fall (p<0.02). CONCLUSION: Smoking rates in young type 1 diabetic patients appear to have fallen during the last decade, and reporting of smoking behaviour is now more honest. PMID- 11423600 TI - Lung carcinoid related Cushing's syndrome: report of three cases and review of the literature. AB - Three patients with lung carcinoid related Cushing's syndrome (LCRCS) treated at Frenchay Hospital, Bristol between 1984 and 1994 are described. The first patient presented with hyperpigmentation 13 years after bilateral adrenalectomy. The second patient had no recurrence or metastases 14 years after removal of a typical carcinoid tumour. The last patient survived nine years after diagnosis of liver metastasis. The possibility of LCRCS should be considered in every patient proved to have Cushing's disease and bilateral adrenal enlargement on abdominal computed tomography. Biochemical sets of investigation (for example, adrenocorticotrophic hormone (ACTH) stimulation, dexamethasone suppression, and metyrapone response) could be misleading and should not be relied upon solely. Search for an ectopic ACTH source should be called off only when ACTH has been demonstrated in the surgically removed specimen, and most importantly, when the serum ACTH concentration returns to normal after surgery. Lung carcinoid tumours are compatible with long survival, and liver metastasis could prove indolent and slowly growing. PMID- 11423601 TI - Parathyroid crisis in a 20 year old--an unusual cause of hypercalcaemic crisis. AB - Since the advent of automated serum analysis, patients with primary hyperparathyroidism (PHPT) are often asymptomatic at presentation or have mild symptoms attributable to the disease. Parathyroid crisis is a rare and potentially fatal complication of PHPT in which patients develop severe hypercalcaemia with signs and symptoms of multiple organ dysfunction. A case of parathyroid crisis in a 20 year old man who presented with brown tumours and renal stones is described. PMID- 11423602 TI - A complicated case of von Hippel-Lindau disease. PMID- 11423603 TI - An unusual cause of paraplegia. PMID- 11423604 TI - Palpable breast mass in a lactating woman. PMID- 11423605 TI - A patient with severe hyperphosphataemia. PMID- 11423606 TI - Abnormal behaviour in a man with massive, generalised, peripheral lymphadenopathy. PMID- 11423607 TI - Oligoarthritis in an elderly woman with diarrhoea and weight loss. PMID- 11423608 TI - A reversible cause of acute renal failure. PMID- 11423616 TI - Coupled transcription and translation within nuclei of mammalian cells. AB - It is widely assumed that the vital processes of transcription and translation are spatially separated in eukaryotes and that no translation occurs in nuclei. We localized translation sites by incubating permeabilized mammalian cells with [3H]lysine or lysyl-transfer RNA tagged with biotin or BODIPY; although most nascent polypeptides were cytoplasmic, some were found in discrete nuclear sites known as transcription "factories." Some of this nuclear translation also depends on concurrent transcription by RNA polymerase II. This coupling is simply explained if nuclear ribosomes translate nascent transcripts as those transcripts emerge from still-engaged RNA polymerases, much as they do in bacteria. PMID- 11423617 TI - Haplotype diversity and linkage disequilibrium at human G6PD: recent origin of alleles that confer malarial resistance. AB - The frequencies of low-activity alleles of glucose-6-phosphate dehydrogenase in humans are highly correlated with the prevalence of malaria. These "deficiency" alleles are thought to provide reduced risk from infection by the Plasmodium parasite and are maintained at high frequency despite the hemopathologies that they cause. Haplotype analysis of "A-" and "Med" mutations at this locus indicates that they have evolved independently and have increased in frequency at a rate that is too rapid to be explained by random genetic drift. Statistical modeling indicates that the A- allele arose within the past 3840 to 11,760 years and the Med allele arose within the past 1600 to 6640 years. These results support the hypothesis that malaria has had a major impact on humans only since the introduction of agriculture within the past 10,000 years and provide a striking example of the signature of selection on the human genome. PMID- 11423618 TI - Clinical resistance to STI-571 cancer therapy caused by BCR-ABL gene mutation or amplification. AB - Clinical studies with the Abl tyrosine kinase inhibitor STI-571 in chronic myeloid leukemia demonstrate that many patients with advanced stage disease respond initially but then relapse. Through biochemical and molecular analysis of clinical material, we find that drug resistance is associated with the reactivation of BCR-ABL signal transduction in all cases examined. In six of nine patients, resistance was associated with a single amino acid substitution in a threonine residue of the Abl kinase domain known to form a critical hydrogen bond with the drug. This substitution of threonine with isoleucine was sufficient to confer STI-571 resistance in a reconstitution experiment. In three patients, resistance was associated with progressive BCR-ABL gene amplification. These studies provide evidence that genetically complex cancers retain dependence on an initial oncogenic event and suggest a strategy for identifying inhibitors of STI 571 resistance. PMID- 11423619 TI - AIDS. Resource needs for HIV/AIDS. PMID- 11423620 TI - White dwarfs and dark matter. PMID- 11423621 TI - Genetic engineering and water. PMID- 11423622 TI - Natural disasters. Texas Medical Center staggered by deadly tropical storm. PMID- 11423624 TI - Neutrino physics. Polymorphous particles solve solar mystery. PMID- 11423623 TI - Human subjects. Volunteer's death prompts review. PMID- 11423625 TI - U.S. research budget. Picture brightens a bit as first bills advance. PMID- 11423626 TI - U.S. science policy. Memo to Congress: get better advice. PMID- 11423627 TI - Cosmology. Math trick may cause tension headache. PMID- 11423628 TI - Astronomy. Cluster watchers view a hot, violent birth. PMID- 11423629 TI - Cancer research. Why some leukemia cells resist STI-571. PMID- 11423630 TI - Neuroscience. Synchronizing the brain's signals. PMID- 11423631 TI - MAP launch. Precision cosmology takes flight. PMID- 11423633 TI - MAP launch. Shaping a universe. PMID- 11423632 TI - MAP launch. Peering backward to the cosmos's fiery birth. PMID- 11423634 TI - Defense research. U.K. agency spawns private high-tech behemoth. PMID- 11423635 TI - Fusion science. Canada bids to host international reactor. PMID- 11423637 TI - Glaciology. Tracking icebergs for clues to climate change. PMID- 11423636 TI - American Geophysical Union meeting. Of ozone, teapots, and snowballs. PMID- 11423638 TI - Communication. Scientific collaborations at a distance. PMID- 11423639 TI - Paleoclimate. Hydrological changes in Africa. PMID- 11423640 TI - Archaeology. On maize and the sunflower. PMID- 11423641 TI - Climate change. Where has all the carbon gone? PMID- 11423642 TI - Microbiology. The great escape. PMID- 11423643 TI - Genomics. On choosing mammalian genomes for sequencing. PMID- 11423644 TI - Neurobiology. Total recall-the memory of addiction. PMID- 11423645 TI - The endless race between plant and pathogen. PMID- 11423646 TI - Finding new ways to fight plant diseases. PMID- 11423647 TI - The push to pit genomics against fungal pathogens. PMID- 11423648 TI - Florida fights to stop citrus canker. PMID- 11423649 TI - Portrait of a pathogen. PMID- 11423650 TI - RNA silencing in plants--defense and counterdefense. AB - RNA silencing is a remarkable type of gene regulation based on sequence-specific targeting and degradation of RNA. The term encompasses related pathways found in a broad range of eukaryotic organisms, including fungi, plants, and animals. In plants, it serves as an antiviral defense, and many plant viruses encode suppressors of silencing. The emerging view is that RNA silencing is part of a sophisticated network of interconnected pathways for cellular defense, RNA surveillance, and development and that it may become a powerful tool to manipulate gene expression experimentally. PMID- 11423651 TI - Evolutionary dynamics of plant R-genes. AB - Plant R-genes involved in gene-for-gene interactions with pathogens are expected to undergo coevolutionary arms races in which plant specificity and pathogen virulence continually adapt in response to each other. Lending support to this idea, the solvent-exposed amino acid residues of leucine-rich repeats, a region of R-genes involved in recognizing pathogens, often evolve at unusually fast rates. But within-species polymorphism is also common in R-genes, implying that the adaptive substitution process is not simply one of successive selective sweeps. Here we document these features in available data and discuss them in light of the evolutionary dynamics they likely reflect. PMID- 11423652 TI - Common and contrasting themes of plant and animal diseases. AB - Recent studies in bacterial pathogenesis reveal common and contrasting mechanisms of pathogen virulence and host resistance in plant and animal diseases. This review presents recent developments in the study of plant and animal pathogenesis, with respect to bacterial colonization and the delivery of effector proteins to the host. Furthermore, host defense responses in both plants and animals are discussed in relation to mechanisms of pathogen recognition and defense signaling. Future studies will greatly add to our understanding of the molecular events defining host-pathogen interactions. PMID- 11423653 TI - Spike transmission and synchrony detection in networks of GABAergic interneurons. AB - The temporal pattern and relative timing of action potentials among neocortical neurons may carry important information. However, how cortical circuits detect or generate coherent activity remains unclear. Using paired recordings in rat neocortical slices, we found that the firing of fast-spiking cells can reflect the spiking pattern of single-axon pyramidal inputs. Moreover, this property allowed groups of fast-spiking cells interconnected by electrical and gamma aminobutyric acid (GABA)-releasing (GABAergic) synapses to detect the relative timing of their excitatory inputs. These results indicate that networks of fast spiking cells may play a role in the detection and promotion of synchronous activity within the neocortex. PMID- 11423654 TI - Liquid crystal alignment on carbonaceous surfaces with orientational order. AB - We used near-edge x-ray absorption fine structure (NEXAFS) spectroscopy to link the orientational bond order at three carbonaceous surfaces-rubbed polyimide, ion beam-irradiated polyimide, and ion beam-irradiated diamondlike carbon films-with the direction of liquid crystal (LC) alignment on these surfaces. We show that, in general, LC alignment can be created on any carbonaceous substrate by inducing orientational order at its surface. Our results form the scientific basis for LC alignment layers consisting of amorphous carbon films in which orientational order near the surface is induced by a directional low-energy ion beam. PMID- 11423655 TI - Conductance switching in single molecules through conformational changes. AB - We tracked over time the conductance switching of single and bundled phenylene ethynylene oligomers isolated in matrices of alkanethiolate monolayers. The persistence times for isolated and bundled molecules in either the ON or OFF switch state range from seconds to tens of hours. When the surrounding matrix is well ordered, the rate at which the inserted molecules switch is low. Conversely, when the surrounding matrix is poorly ordered, the inserted molecules switch more often. We conclude that the switching is a result of conformational changes in the molecules or bundles, rather than electrostatic effects of charge transfer. PMID- 11423656 TI - A 14,000-year oxygen isotope record from diatom silica in two alpine lakes on Mt. Kenya. AB - Oxygen isotopes are sensitive tracers of climate change in tropical regions. Abrupt shifts of up to 18 per mil in the oxygen isotope ratio of diatom silica have been found in a 14,000-year record from two alpine lakes on Mt. Kenya. Interpretation of tropical-montane isotope records is controversial, especially concerning the relative roles of precipitation and temperature. Here, we argue that Holocene variations in delta(18)O are better explained by lake moisture balance than by temperature-induced fractionation. Episodes of heavy convective precipitation dated approximately 11,100 to 8600, 6700 to 5600, 2900 to 1900, and <1300 years before the present were linked to enhanced soil erosion, neoglacial ice advances, and forest expansion on Mt. Kenya. PMID- 11423657 TI - Paleobotanical evidence for near present-day levels of atmospheric Co2 during part of the tertiary. AB - Understanding the link between the greenhouse gas carbon dioxide (CO(2)) and Earth's temperature underpins much of paleoclimatology and our predictions of future global warming. Here, we use the inverse relationship between leaf stomatal indices and the partial pressure of CO(2) in modern Ginkgo biloba and Metasequoia glyptostroboides to develop a CO(2) reconstruction based on fossil Ginkgo and Metasequoia cuticles for the middle Paleocene to early Eocene and middle Miocene. Our reconstruction indicates that CO(2) remained between 300 and 450 parts per million by volume for these intervals with the exception of a single high estimate near the Paleocene/Eocene boundary. These results suggest that factors in addition to CO(2) are required to explain these past intervals of global warmth. PMID- 11423658 TI - Quinones as the redox signal for the arc two-component system of bacteria. AB - The Arc two-component signal transduction system mediates adaptive responses of Escherichia coli to changing respiratory conditions of growth. Under anaerobic conditions, the ArcB sensor kinase autophosphorylates and then transphosphorylates ArcA, a global transcriptional regulator that controls the expression of numerous operons involved in respiratory or fermentative metabolism. We show that oxidized forms of quinone electron carriers act as direct negative signals that inhibit autophosphorylation of ArcB during aerobiosis. Thus, the Arc signal transduction system provides a link between the electron transport chain and gene expression. PMID- 11423659 TI - Consistent land- and atmosphere-based U.S. carbon sink estimates. AB - For the period 1980-89, we estimate a carbon sink in the coterminous United States between 0.30 and 0.58 petagrams of carbon per year (petagrams of carbon = 10(15) grams of carbon). The net carbon flux from the atmosphere to the land was higher, 0.37 to 0.71 petagrams of carbon per year, because a net flux of 0.07 to 0.13 petagrams of carbon per year was exported by rivers and commerce and returned to the atmosphere elsewhere. These land-based estimates are larger than those from previous studies (0.08 to 0.35 petagrams of carbon per year) because of the inclusion of additional processes and revised estimates of some component fluxes. Although component estimates are uncertain, about one-half of the total is outside the forest sector. We also estimated the sink using atmospheric models and the atmospheric concentration of carbon dioxide (the tracer-transport inversion method). The range of results from the atmosphere-based inversions contains the land-based estimates. Atmosphere- and land-based estimates are thus consistent, within the large ranges of uncertainty for both methods. Atmosphere based results for 1980-89 are similar to those for 1985-89 and 1990-94, indicating a relatively stable U.S. sink throughout the period. PMID- 11423660 TI - Changes in forest biomass carbon storage in China between 1949 and 1998. AB - The location and mechanisms responsible for the carbon sink in northern mid latitude lands are uncertain. Here, we used an improved estimation method of forest biomass and a 50-year national forest resource inventory in China to estimate changes in the storage of living biomass between 1949 and 1998. Our results suggest that Chinese forests released about 0.68 petagram of carbon between 1949 and 1980, for an annual emission rate of 0.022 petagram of carbon. Carbon storage increased significantly after the late 1970s from 4.38 to 4.75 petagram of carbon by 1998, for a mean accumulation rate of 0.021 petagram of carbon per year, mainly due to forest expansion and regrowth. Since the mid 1970s, planted forests (afforestation and reforestation) have sequestered 0.45 petagram of carbon, and their average carbon density increased from 15.3 to 31.1 megagrams per hectare, while natural forests have lost an additional 0.14 petagram of carbon, suggesting that carbon sequestration through forest management practices addressed in the Kyoto Protocol could help offset industrial carbon dioxide emissions. PMID- 11423661 TI - The epidemic behavior of the hepatitis C virus. AB - Hepatitis C virus (HCV) is a leading worldwide cause of liver disease. Here, we use a new model of HCV spread to investigate the epidemic behavior of the virus and to estimate its basic reproductive number from gene sequence data. We find significant differences in epidemic behavior among HCV subtypes and suggest that these differences are largely the result of subtype-specific transmission patterns. Our model builds a bridge between the disciplines of population genetics and mathematical epidemiology by using pathogen gene sequences to infer the population dynamic history of an infectious disease. PMID- 11423662 TI - A protein antibiotic in the phage Qbeta virion: diversity in lysis targets. AB - A(2), a capsid protein of RNA phage Qbeta, is also responsible for host lysis. A(2) blocked synthesis of murein precursors in vivo by inhibiting MurA, the catalyst of the committed step of murein biosynthesis. An A(2)-resistance mutation mapped to an exposed surface near the substrate-binding cleft of MurA. Moreover, purified Qbeta virions inhibited wild-type MurA, but not the mutant MurA, in vitro. Thus, the two small phages characterized for their lysis strategy, Qbeta and the small DNA phage phiX174, effect host lysis by targeting different enzymes in the multistep, universally conserved pathway of cell wall biosynthesis. PMID- 11423663 TI - Recruitment of HAT complexes by direct activator interactions with the ATM related Tra1 subunit. AB - Promoter-specific recruitment of histone acetyltransferase activity is often critical for transcriptional activation. We present a detailed study of the interaction between the histone acetyltransferase complexes SAGA and NuA4, and transcription activators. We demonstrate by affinity chromatography and photo cross-linking label transfer that acidic activators directly interact with Tra1p, a shared subunit of SAGA and NuA4. Mutations within the COOH-terminus of Tra1p disrupted its interaction with activators and resulted in gene-specific transcriptional defects that correlated with lowered promoter-specific histone acetylation. These data demonstrate that the essential Tra1 protein serves as a common target for activators in both SAGA and NuA4 acetyltransferases. PMID- 11423664 TI - Requirement of ERK activation for visual cortical plasticity. AB - Experience-dependent plasticity in the developing visual cortex depends on electrical activity and molecular signals involved in stabilization or removal of inputs. Extracellular signal-regulated kinase 1,2 (also called p42/44 mitogen activated protein kinase) activation in the cortex is regulated by both factors. We show that two different inhibitors of the ERK pathway suppress the induction of two forms of long-term potentiation (LTP) in rat cortical slices and that their intracortical administration to monocularly deprived rats prevents the shift in ocular dominance towards the nondeprived eye. These results demonstrate that the ERK pathway is necessary for experience-dependent plasticity and for LTP of synaptic transmission in the developing visual cortex. PMID- 11423665 TI - Intracellular anions as the voltage sensor of prestin, the outer hair cell motor protein. AB - Outer hair cells (OHCs) of the mammalian cochlea actively change their cell length in response to changes in membrane potential. This electromotility, thought to be the basis of cochlear amplification, is mediated by a voltage sensitive motor molecule recently identified as the membrane protein prestin. Here, we show that voltage sensitivity is conferred to prestin by the intracellular anions chloride and bicarbonate. Removal of these anions abolished fast voltage-dependent motility, as well as the characteristic nonlinear charge movement ("gating currents") driving the underlying structural rearrangements of the protein. The results support a model in which anions act as extrinsic voltage sensors, which bind to the prestin molecule and thus trigger the conformational changes required for motility of OHCs. PMID- 11423666 TI - National dialogue on cancer: a bold new effort. PMID- 11423667 TI - CEOs consider ways to intensify the battle against cancer. PMID- 11423668 TI - The oncologic four-minute mile. PMID- 11423669 TI - STI571: targeting BCR-ABL as therapy for CML. AB - Therapeutic agent STI571 (signal transduction inhibitor number 571) is a rationally developed, potent, and selective inhibitor for abl tyrosine kinases, including bcr-abl, as well c-kit and the platelet-derived growth factor receptor tyrosine kinases. Results of clinical trials to date have demonstrated the crucial role of the bcr-abl tyrosine kinase in chronic myelogenous leukemia (CML) pathogenesis and the potential of anticancer agents designed to target specific molecular abnormalities in human cancer. An initial phase I study of STI571 included 83 Ph(+) CML patients who had failed interferon-based therapy. Patients were required to be in chronic phase, defined liberally as less than 15% blasts in blood or bone marrow. Patients were treated with once-daily oral doses of STI571 in 14 successive dose cohorts ranging from 25-1,000 mg. In this phase I study, no dose-limiting toxicity was encountered and toxicity at all dose levels was minimal. The threshold for a maximally effective dose was found at 300 mg; for patients treated at or above this level, complete hematologic response was seen in 98% of patients, with complete cytogenetic responses in 13% and major cytogenetic responses in 31%. With a median duration of follow-up of 310 days, ongoing responses are evident in 96% of patients. In the phase II study of the accelerated phase of CML, 233 patients were treated with either 400 or 600 mg of STI571. With similar follow-up to the chronic phase trial, 91% of patients showed a hematological response; 63% of patients achieved a complete hematological response but not all patients had recovery of peripheral blood counts. In addition to the phase II clinical trials with STI571, a phase III trial randomizing newly diagnosed patients to either interferon with low-dose s.c. cytosine arabinoside versus STI571 is ongoing; this trial accrued rapidly and data collection is ongoing. Integration of STI571 into CML treatment algorithms will require long-term follow-up data from the ongoing phase II and III clinical studies. PMID- 11423670 TI - Thymoma: update for the new millennium. AB - Thymomas are relatively common tumors of the anterior superior mediastinum. They are usually relatively slowly growing tumors and their prognosis depends on the macroscopic and microscopic invasion of surrounding tissues. Surgery is the mainstay treatment of thymomas, and complete resection represents one of the most important prognostic factors in this disease. Other important prognostic indicators include the tumor stage and size and the presence of symptoms. Postoperative radiotherapy is indicated in tumors with invasion of surrounding tissues, even if resection was radical, since it improves local control and survival. Cytotoxic chemotherapy has been employed in several relatively small phase II studies and in advanced disease has been demonstrated to produce a 50% 80% objective response rate. Neoadjuvant cytotoxic chemotherapy and/or external beam radiotherapy has been used with some success in patients with tumors which are not readily resectable. Novel antiproliferative systemic agents, with both cytotoxic and cytostatic mechanisms of action, are being tested in ongoing prospective clinical trials. PMID- 11423671 TI - High-dose therapy in lymphomas: a review of the current status of allogeneic and autologous stem cell transplantation in Hodgkin's disease and non-Hodgkin's lymphoma. AB - Autologous stem cell transplantation has proven to be beneficial in selected patients with Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL). In patients with HD, transplantation appears to increase event-free survival in patients who fail to enter complete remission with initial therapy. When a patient relapses after a complete remission, transplantation is probably the best option and particularly so if the remission lasted less than 1 year. Transplantation as part of primary therapy for very high-risk patients may be beneficial, but is not standard therapy at this time. For patients with diffuse large-cell NHL, transplantation can be considered standard therapy for relapsed patients if they have chemotherapy-sensitive disease. The use of transplantation for high-risk patients in complete remission is promising, but definite recommendations cannot be made at this time. For follicular lymphomas, selected patients seem to benefit and studies are ongoing. Finally, the use of allogeneic stem cell transplantation can be useful in a select group of younger patients. PMID- 11423672 TI - Review of recent advances in fluorescence bronchoscopy in early localization of central airway lung cancer. AB - Centrally located lung cancers are radiologically occult until so far advanced as to have a low cure rate or require extensive resection for cure, but at a cost of high morbidity. These cancers represent about one-fifth of new lung cancers. Autofluorescence bronchoscopy appears to be an important tool in localizing premalignant and early malignant lesions in the large central airways, particularly when applied to high-risk patients. Applications include studies of molecular biology of premalignancy and early malignancy, chemoprevention studies, endobronchial therapy studies, localization of synchronous tumors, estimation of the extent of field cancerization, and better estimation of resection margins. Autofluorescence bronchoscopy appears to be significantly more sensitive than white light examination but has low specificity. This technology is likely to gain widespread use when evaluation of sputum for malignant changes is both more sensitive and specific, and when its application is demonstrated to reduce mortality in this important subgroup of non-small cell lung cancer patients. PMID- 11423673 TI - Molecular genetics of neuroblastoma and the implications for clinical management: a review of the MSKCC experience. AB - Neuroblastoma (NB) is a biological, genetic, and morphological heterogeneous neoplasm and demonstrates diverse clinical behavior. There exist at least three clinical patterns of NB: A) spontaneously regressing widespread disease; B) not metastatic local-regional disease, and C) metastatic disease (stage 4), frequently with lethal consequences. Patients with non-stage 4 NB are expected to survive even without medical treatment whereas stage 4 patients have an overall survival rate of 20% despite multimodality therapy protocols. The clinical management of patients with NB is therefore challenged by the objective identification of cases in which noncytotoxic approaches can be safely taken. Experience in the last decade at Memorial Sloan-Kettering Cancer Center supports the hypothesis that the natural history of disease defines relevant clinical groups of NB and has distinct molecular genetic profiles allowing therapeutic approaches tailored for each group. Here we review the natural history and clinicobiological features of 113 NB cases managed uniformly in our institution in an attempt to characterize useful genetic markers to support the decision making of noncytotoxic versus cytotoxic approaches for each category of NB. PMID- 11423674 TI - Activity and safety of vinorelbine combined with doxorubicin or fluorouracil as first-line therapy in advanced breast cancer: a stratified phase II study. AB - PURPOSE: To evaluate the efficacy and safety of vinorelbine combined with doxorubicin or continuous infusion of fluorouracil as initial therapy for advanced breast cancer. AUBJECTS AND METHODS: A total of 118 women who had not received chemotherapy for advanced breast cancer were enrolled and included in the intent-to-treat analysis. Subjects were stratified into two treatment groups. If subjects were candidates for anthracycline therapy, they received doxorubicin 50 mg/m(2) on day 1 and vinorelbine 25 mg/m(2) on days 1 and 8 (n = 62). If subjects had received adjuvant anthracycline therapy or had cardiac disease, they received fluorouracil 750 mg/m(2)/day by continuous infusion on days 1 through 5 and vinorelbine 30 mg/m(2) on days 1 and 5 (n = 56). The regimens were repeated every 21 days until evidence of progression of disease or severe toxicity. RESULTS: For doxorubicin and vinorelbine, the objective response rate was 55% (95% confidence interval [CI]: 42% to 68%), median time to disease progression was 34 weeks, median time to treatment failure was 32 weeks, and median survival was 92 weeks (95% CI: 72 to 128 weeks). For fluorouracil and vinorelbine, the objective response rate was 45% (95% CI: 31% to 59%), median time to disease progression was 32 weeks, median time to treatment failure was 30 weeks, and median survival was 53 weeks (95% CI: 47 to 64 weeks). The most common adverse event was grade 3 or 4 granulocytopenia, which occurred in 95% of subjects in the doxorubicin-vinorelbine group and in 88% of those in the fluorouracil-vinorelbine group. The most common nonhematologic adverse event was grade 3 or 4 stomatitis, which occurred in 9% and 32% of subjects in the two groups, respectively. CONCLUSION: Both vinorelbine-containing regimens appear to offer useful options as initial therapy for advanced breast cancer. Both regimens were active, and any efficacy differences between the two may have been related to differences in prognosis for the anthracycline-pretreated group (i.e., selection for prior aggressive adjuvant therapy) and or comorbid cardiac conditions. Both regimens were associated with predictable but manageable toxicity, but a lower dose of fluorouracil (e.g., 600 mg/m(2)/day) should be used to reduce the risk of stomatitis. PMID- 11423675 TI - Osteoporosis in survivors of acute lymphoblastic leukemia. AB - Osteoporosis is currently receiving increasing attention as an important late effect in survivors of childhood cancer and its treatment because of their quality of life and its negative effect on the survivors' ability to perform developmentally appropriate activities. Survivors of childhood cancer are especially vulnerable because they are affected during childhood and adolescence, a time when peak bone mass should be achieved. This paper reviews decreased bone density in acute lymphoblastic leukemia (ALL), which is the most common childhood cancer and has a cure rate approaching 80%. Osteopenia/osteoporosis has been observed in all phases of the disease: at diagnosis, during treatment, and throughout the post-treatment period for as long as 20 years. Among the findings that have been described are musculoskeletal pain, disturbed gait, fractures, kyphosis, lordosis, and growth failure. Risk factors not specifically related to ALL include smoking, ingestion of carbonated beverages, and family history of "brittle bone" or fractures. Patients should be counseled in regard to diet, exercise, smoking cessation, and avoidance of carbonated beverages. There are a number of options for specific drug therapy; however, the administration of bisphosponates to children and adolescents must be approached with caution. Research is needed to determine how extensive the problem is and how to best prevent and treat the osteopenia/osteoporosis associated with ALL. PMID- 11423676 TI - Losing God. AB - Shortly before his death in 1995, Kenneth B. Schwartz, a cancer patient at Massachusetts General Hospital (MGH), founded the Kenneth B. Schwartz Center. The Schwartz Center is a non-profit organization dedicated to supporting and advancing compassionate health care delivery, which provides hope to the patient, support to caregivers, and sustenance to the healing process. The center sponsors the Schwartz Center Rounds, a monthly multidisciplinary forum where caregivers reflect on important psychosocial issues faced by patients, their families, and their caregivers, and gain insight and support from fellow staff members. Nebulous language, distrust, and dogma confound spiritual aspects of cancer care. However, existential well being is an important determinant of quality of life: finding meaning and purpose make suffering more tolerable. The case presented is of a patient who experienced "losing God" as a Hodgkin's disease survivor with metastatic prostate cancer and severe coronary artery disease. His caregivers were able to provide the sense of community in which he could re-establish his faith. Health care providers do not have to be religious in order to help patients to deal with a spiritual crisis. The clinical skills of compassion need to be deployed to diagnose and respond to spiritual suffering. Acknowledging and addressing anger or guilt, common sources of suffering, are essential to adjustment. Simply being there for the patient and being open to their hurt can help resolve their spiritual crisis, a responsibility that is shared by the whole health care team. PMID- 11423677 TI - The molecular perspective: ultraviolet light and pyrimidine dimers. PMID- 11423679 TI - Capitol report. PMID- 11423681 TI - The "foul disease" and privacy: the effects of venereal disease and patient demand on the medical marketplace in early modern London. AB - This article examines medical advertisements for venereal disease treatment from late Stuart London. It explores how privacy issues influenced the services provided by early modern venereologists. It shows that practitioners who sought to get ahead in the competitive field of venereology began to offer private treatment at a time when other physicians seem not to have provided that service. Therefore, market forces such as patient demand had an innovatory effect on early modern medical ethics. The same dynamic that caused venereal patients to seek privacy also led them to demand a practitioner of their own sex. Infected women clearly wished to be treated by a female practitioner. Many male practitioners forged partnerships with women in order to attract female clientele. These partnerships were frequently based on familial connections, most often between husband and wife. The presence of widespread VD in London helped sustain a sizable number of female practitioners who specialized in venereology. PMID- 11423682 TI - "A host of experienced microscopists": the establishment of histology in nineteenth-century Edinburgh. AB - Edinburgh was the foremost center in Britain for the introduction of microscopic anatomy into medical training. It therefore offers an instructive case study of the way in which what was initially an obscure and exotic technology eventually became a regular part of medical education. The paper explores the process by which skills that were originally the preserve of a small number of pioneers in histology came to be transmitted to a wider population. It focuses, in particular, on the transition from an authoritarian style of pedagogy, best exemplified by the histological teaching of John Hughes Bennett, to the more collegial styles of interaction between microscopists that came to be embodied in the Edinburgh Physiological Society. PMID- 11423683 TI - Train wrecks to typhoid fever: the development of railroad medicine organizations, 1850-World War I. AB - From their beginning American railroads developed medical arrangements to care for the large number of workers and passengers they killed or injured. After the Civil War, both labor unrest and liability concerns led them to expand and formalize these arrangements, and three forms of organization arose. Western roads, facing an almost complete lack of medical facilities, developed employee funded hospital organizations. In the east, companies created medical organizations under a salaried chief surgeon and contracted with local physicians to provide care. A third model, pioneered by the Baltimore & Ohio Railroad in the 1880s, devised a beneficial society that provided medical care and compensation for injured workers. Although these organizations involved both contract practice and group hospitalization, the AMA seems to have ignored them. In the 1880s railroad physicians developed their own organizations, including the National Association of Railway Surgeons, in which they discussed problems of professionalization and such medical matters as "railway spine." Concern with costs and labor turnover also led the carriers into preventive medicine. Some roads provided smallpox and typhoid vaccinations, campaigned against malaria, improved passenger-car sanitation, required physical examinations of their employees, and trained them in first aid. By World War I, railroad medical organizations provided care to nearly two million employees and employment to about 10 percent of all physicians. PMID- 11423684 TI - The expurgation of medical books in sixteenth-century Spain. AB - Medical books figured prominently on lists of scientific books censured or expurgated by ecclesiastic authorities in the second half of the sixteenth century. A systematic examination of this censorship is still wanting. The following study, which describes and explains the mechanism of expurgation of centuria iv in Amatus Lusitanus's description of medical cases (Lyons edition, 1580), is an example of the sort of questions and information that can be drawn from such an examination. In particular, the expurgation of Amatus's discussion of false conception suffered by a nun is analyzed. Amatus did not rule out the possibility of natural virginal pregnancy, and in doing so he relied on medical authorities (Averroes) as well as on a Hebrew rabbinical source (Alphabet of Ben Sira). PMID- 11423685 TI - Parathyroid hormone-related protein as a renal regulating factor. From vessels to glomeruli and tubular epithelium. AB - Parathyroid hormone (PTH) and PTH-related protein (PTHrP) produce similar biological effects through the PTH/PTHrP receptor. Less is known about the physiological role of PTHrP, which was first identified as the agent of the humoral hypercalcemia of malignancy. Despite the widespread production of PTHrP in healthy individuals, the concentration of the protein is below the detectable limit of current assays, suggesting that PTHrP normally functions locally in an autocrine or paracrine manner. Thus, some differences in their biological activities have been described and they may be related to the presence of different receptors. In this regard, a second receptor that binds selectively to PTH has also been found. Recent studies have demonstrated the expression of both PTH/PTHrP receptor and protein in the renal glomeruli. Moreover, there are convincing data that support a direct role of PTH and PTHrP in modulating renal blood flow and glomerular filtration rate. This multifunctional protein, PTHrP, also has a proliferative effect on both glomerular mesangial cells and tubular epithelial cells. Increases in the expression of PTHrP have been observed in several experimental models of nephropathies, suggesting that PTHrP upregulation is a common event associated with the mechanism of renal injury and repair. PMID- 11423686 TI - The BsmI vitamin D receptor gene polymorphism in Israeli populations and in perimenopausal and osteoporotic Ashkenazi women. AB - BACKGROUND: The association between vitamin D receptor (VDR) gene polymorphisms and bone mineral density (BMD) is controversial, and may be effected by ethnic ancestry and age. AIMS: To determine the distribution of the BsmI VDR gene polymorphism in healthy Israeli populations, and to study its association with BMD in perimenopausal and osteoporotic Ashkenazi women. METHODS: Allele and genotype frequencies of the VDR gene defined by BsmI restriction site were determined in 634 healthy Israelis of seven ethnic groups, 90 Ashkenazi perimenopausal women and in 75 Ashkenazi osteoporotic women. Genotype-related differences in spinal and femoral neck BMD were determined in Ashkenazi perimenopausal women. Allele and genotype frequencies in Ashkenazi osteoporotic women were compared with Ashkenazi controls. RESULTS: The frequency of the BB genotype was higher in Yemenites compared with Ashkenazi and Libyan Jews (23, 11 and 8%, respectively, p < 0.05), and lower in Ashkenazi compared with Iraqi and Persian Jews (11, 20 and 21%, respectively, p = 0.05). BMD did not vary by genotype in perimenopausal women, nor were there differences in the frequencies of the B allele or the BB genotype in osteoporotic women compared with controls. CONCLUSIONS: There is ethnic variability in the frequency of the BsmI VDR gene polymorphism. In Ashkenazi perimenopausal and osteoporotic women this polymorphism is not associated with BMD. PMID- 11423687 TI - Peritoneal sodium mass removal in continuous ambulatory peritoneal dialysis and automated peritoneal dialysis: influence on blood pressure control. AB - BACKGROUND/AIM: Sodium and water retention is common in peritoneal dialysis patients and contributes to cardiovascular disease. As peritoneal sodium removal depends partly on dwell time, and automated peritoneal dialysis (APD) often uses short dwell time exchanges, the aim of this study was to compare the 24-hour peritoneal sodium removal in APD and standard continuous ambulatory peritoneal dialysis (CAPD) patients and to analyze its possible influence on blood pressure control. METHODS: A total of 53 sodium balance studies (30 in APD and 23 in CAPD) were performed in 36 stable peritoneal dialysis patients. The 24-hour net removal of sodium was calculated as follows: M = ViCi - VdCd, where Vd is the 24-hour drained volume, Cd is the solute sodium concentration in Vd, Vi is the amount of solution used during a 24-hour period, and Ci is the sodium concentration in Vi. Peritoneal sodium removal was compared between APD and CAPD patients. Residual renal function, serum sodium concentration, daily urinary sodium losses, weekly peritoneal Kt/V and creatinine clearance, 4-hour dialysate/plasma creatinine ratio, proportion of hypertonic solutions, net ultrafiltration, systolic and diastolic blood pressures, and need for antihypertensive therapy were also compared between the groups. RESULTS: Peritoneal sodium removal was higher (p < 0.001) in CAPD than in APD patients. There were no significant differences in residual renal function, serum sodium concentration, urinary sodium losses, peritoneal urea or creatinine clearances, 4-hour dialysate/plasma creatinine ratio, or proportion of hypertonic solutions between groups. The net ultrafiltration was higher in CAPD patients and correlated strongly (r = 0.82; p < 0.001) with peritoneal sodium removal. In APD patients, peritoneal sodium removal increased significantly only in those patients with a second daytime exchange. The systolic blood pressure was higher (p < 0.05) in APD patients, and the proportion of patients with antihypertensive therapy was also higher in APD patients, although no significant relationship between blood pressure values and amount of peritoneal sodium removal was found. CONCLUSIONS: The 24-hour sodium removal is higher in CAPD than in APD patients, and there is a trend towards better hypertension control in CAPD patients. As hypertension control and volume status are important indices of peritoneal dialysis adequacy, our results have to be considered in the choice of the peritoneal dialysis modality. PMID- 11423688 TI - Lack of effect of chronic hepatitis C virus infection on T-cell cytokine production in chronic hemodialysis patients. AB - It has been shown that chronic hemodialysis modifies, to some extent, the normal immune response by both T and B lymphocytes elicited by antigenic stimulation, e.g. by impairing the T-cell-dependent response after vaccination. A new technique, i.e. flow cytometry, enables to assess intracytoplasmically, at the single cell level, the production of a given cytokine. By using it, we studied in healthy volunteers (HV) and in chronic hemodialysis (CHD) patients, with respect to their hepatitis C virus (HCV) status, the production by the T lymphocytes of type 1, and type 2 cytokines. We studied the following cytokines (CK): IL-2, IL 4, IL-5, IL-6, IL-10, IFN-gamma and TNF-alpha in the T-cell lymphocytes (whole, CD4+ and CD8+). There were 13 HV and 59 CHD patients (36 HCV(-) and 23 HCV(+)). Amongst the latter, there were 32 men and 27 women, aged 59.5 +/- 2 years, undergoing CHD since 70 +/- 9.4 months. We found that: (1) the total number of lymphocytes as well as those expressing CD3, CD4, or CD19 were significantly decreased in CHD patients as compared to those from HV; (2) the total number of lymphocytes as well as their different subsets were similar in HCV(+) and in HCV( ) CHD patients; (3) the frequency of T-cell-expressing IL-5 or IL-10 was always low (<1%) in both HV and CHD groups; (4) overall in CHD patients, the mean percentages of T lymphocytes expressing IL-2, IL-4, IFN-gamma or TNF-alpha were respectively 31 +/- 13, 2.5 +/- 1.3, 28 +/- 12 and 34 +/- 11% and were not statistically different between HCV(+) and HCV(-) patients; (5) IL-2 was mainly produced by CD4+ T cells, whereas IFN-gamma was produced by CD8+ T cells, in both HV and CHD groups, and (6) the lymphocytes of CHD patients produced significantly more IL-2 and IL-4 than those from HV, suggesting an activation of their T lymphocytes. We conclude that using the cytokine flow cytometry assay, our study demonstrated that in HCV(+) CHD patients, as opposed to what has been described for HCV(+) patients with normal renal function, there was no impairment in the production of type 1 cytokines by peripheral blood mononuclear cells when compared to HCV(-) CHD patients. Conversely to HV, T lymphocytes from CHD patients are activated. PMID- 11423689 TI - Identifying patients at risk for hemodialysis underprescription. AB - Underprescription of hemodialysis is an important barrier to adequate delivery of dialysis. We sought to determine which patient factors are associated with hemodialysis underprescription and to examine variation in prescription across facilities. For 721 randomly selected patients from all 22 chronic hemodialysis units in northeast Ohio, we calculated prescribed Kt/V based on dialyzer urea clearance at prescribed blood and dialysate flow (K), prescribed treatment time (t), and anthropometric volume (V). A minimum 'prescribed Kt/V' of 1.3 has been recommended to ensure an adequate 'delivered Kt/V' of 1.2. Using this criterion, 15% of patients had a low prescribed Kt/V. Prescribed Kt was strongly related to patient anthropometric volume but not to other patient characteristics (age, gender, race, cause of renal failure, number of years on dialysis, number of comorbid conditions). A 10-liter increase in V was associated with an 8.3-liter increase in prescribed Kt. However, a 13-liter increase in prescribed Kt would be needed to maintain a prescribed Kt/V of 1.3. As a result, the proportion of patients with low prescriptions increased from 2% of patients with V <35 liters to 42% of patients with V > or =50 liters. In addition, the prevalence of low prescriptions varied dramatically across facilities (range 0-47%) even after accounting for volumes of individual patients. Of the 109 patients with low prescription, 75% would achieve a prescribed Kt/V of 1.3 with less than 30 min of additional treatment time. In conclusion, large patients and patients at specific facilities are at increased risk for underprescription of hemodialysis. Most patients with low prescriptions would achieve a prescribed Kt/V of 1.3 with a modest increase in treatment time. PMID- 11423690 TI - Evaluation of the severity of traumatic rhabdomyolysis using technetium-99m pyrophosphate scintigraphy. AB - A quantitative scoring method was designed to assess the extent of muscle damage. Technetium-99m pyrophosphate (99mTc-PYP) scintigraphy was performed for 9 patients experiencing crush injury in the Chichi (Taiwan) earthquake. The magnitude of muscle uptake of 99mTc-PYP was graded as follows: grade 0, less than bone radioactivity (BRA); grade 1, equal to BRA; grade 2, higher than BRA; or grade 3, greatly higher than BRA. The area of muscle injury was estimated according to the rule of nines. The sum of the muscle injury size multiplied by its corresponding grading was defined as the anterior or posterior score according to the anterior or posterior images. Each image was interpreted by two physicians and average anterior and posterior scores were calculated. The muscle score was defined as the geometric mean of the average anterior and posterior scores. Significant correlations were obtained between the muscle score and duration of time trapped (r = 0.868, p < 0.01), peak serum creatine kinase level (r = 0.866, p < 0.01), peak serum phosphorus level (r = 0.877, p < 0.01) and number of hospital days (r = 0.875, p < 0.01). A negative correlation between the muscle score and blood pH (r = -0.706, p < 0.01) was also observed. We concluded that this scoring method may be used as an adjunct for evaluating the locations of trauma and the severity of crush syndrome, and for predicting the duration of hospital stay. PMID- 11423691 TI - Self-assessed quality of life in peritoneal dialysis patients. AB - BACKGROUND/AIMS: Studies comparing quality of life (QOL) between peritoneal and hemodialysis patients have yielded inconsistent results. Physical (PCS) and mental component summary (MCS) scales of Short Form 36 (SF-36) health survey are highly validated measures of self-assessed QOL. We sought to evaluate these indices in PD patients: (1) as measures of QOL, (2) predictors of QOL, (3) to study change in QOL over time, and (4) to compare QOL in PD vs. hemodialysis patients. METHODS: SF-36 questionnaires were administered every 3 months to patients over a 2-year period and PCS and MCS were calculated. Mean follow-up was 15.3 +/- 6.6 months for PD and 14.5 +/- 5.7 months for HD. RESULTS: Average PCS in PD (31.8 +/- 7.8) was lower than HD (36.9 +/- 9.8) (p < 0.02), while MCS was similar in the groups (p = NS). The prevalence of depression was 26.1% in PD and 25.4% in HD patients (p = NS). Serum albumin was the only significant predictor of PCS among PD patients and explained much of the decrease in PCS in them. The number of hospitalizations and in-hospital days were significantly lower for PD compared to HD patients (p < 0.05). PCS as well as MCS remained stable in both groups throughout the observation period. CONCLUSION: Self-assessed physical function is diminished, while mental function is similar in PD compared to HD patients. When corrected for serum albumin, this difference is eliminated. Over time, QOL in patients treated with PD remained stable. PMID- 11423692 TI - Access blood flow as a predictor of early failures of native arteriovenous fistulas in hemodialysis patients. AB - Blood flow imaging using color doppler has proven effective in predicting graft failures in hemodialysis patients, but its effect on native arteriovenous fistulas (AVF) is not well known. This study was performed to investigate whether measurements of the access blood flow can be used as predictors of an early failure of a native AVF in hemodialysis patients. Fifty-three consecutive patients who received native AVF operations were included in this study. Access blood flow was measured at 1 week after operations, and AVF function was followed for 4 months. During the follow-up, access failures developed in 10 patients at 9.8 +/- 3.5 weeks. AVF blood flow was significantly lower in the failure group (n = 10) than in the patent group (n = 43) (450 +/- 214 vs. 814 +/- 348 ml/min, p = 0.003). The incidence of access failures was higher in the patients with a flow <350 ml/min (n = 9) compared to the patients with a flow >350 ml/min (n = 44) (55.5 vs. 11.3%, p = 0.008). The diameters of veins were significantly smaller in the failure group than in the patent group (3.5 +/- 0.5 vs. 4.1 +/- 0.7 mm, p = 0.018). The incidence of diabetes mellitus was higher in the failure group than in the patent group (90 vs. 51%, p = 0.025). However, age, sex, duration from an operation to first cannulation, and different AVF sites did not make a significant difference between the two groups. Our data suggest that access blood flow measurements using color doppler ultrasound during early postoperative periods are useful parameters in predicting an early failure of a native AVF in hemodialysis patients. PMID- 11423693 TI - A case review: anticoagulation in hemodialysis patients with heparin-induced thrombocytopenia. AB - We describe a patient with acute renal failure who subsequently developed heparin induced thrombocytopenia (HIT) while on hemodialysis. Heparin was immediately discontinued and alternative modes of anticoagulation were considered as further hemodialysis was indicated. With several options available, a review of the current literature was performed to aid in the selection of the most appropriate method. We conclude that there is currently no ideal hemodialysis anticoagulation agent for a patient with HIT. Among the currently available alternatives, no anticoagulation or regional citrate infusion during hemodialysis appears to be the most reliable and safest option for these patients. Based upon its safety, reversibility, low cost and availability, a trial of warfarin also may be attempted for hemodialysis anticoagulation. Both danapranoid and lepirudin have been used effectively in hemodialysis patients with HIT, but have the disadvantage of prolonged half-lives in patients with renal failure and relatively high cost. PMID- 11423694 TI - Successful treatment of massive lower gastrointestinal bleeding caused by mixed infection of cytomegalovirus and mucormycosis in a renal transplant recipient. AB - We describe a case of lower gastrointestinal bleeding due to mixed infection of cytomegalovirus (CMV) and mucormycosis in a renal transplant recipient. A 33-year old male received renal transplantation and his clinical course was uneventful. On the 18th postoperative day, acute rejection was developed and this was treated with high-dose methylprednisolone and OKT3. During antirejection treatment, sudden onset massive hematochezia was developed. Emergency colonofibroscopy revealed multiple colonic ulcers and pathologic findings were consistent with mucormycosis and CMV infection. The patient was successfully treated with amphotericin B (1.0-1.5 mg/kg) and ganciclovir (62.5-125 mg/day) for 5 weeks. To our knowledge, this is the first report showing coexistence of mucormycosis and CMV in the colon ulcer base. This finding suggests that CMV infection may trigger fungal infection in the pathogenesis of colonic ulcer. PMID- 11423695 TI - Recurrence of light chain nephropathy in a renal allograft. A case report and review of the literature. AB - Myeloma and monoclonal gammopathies can affect the kidney in many ways with cast nephropathy and light chain deposition disease being the most commonly recognised histological entities. Renal transplantation in these patients remains controversial both because of the risk of recurrent disease affecting the graft and also because of concerns around the possibility of disease relapse within the patient. We suggest that the histological pattern of disease within the native kidneys is crucial in the overall assessment of these patients for renal transplantation. Those patients in whom renal deposition of light chains is associated with a proliferative glomerulonephritis have a considerably worse graft survival than those presenting with cast nephropathy. PMID- 11423696 TI - Life-threatening hyperkalemia after intravenous labetolol injection for hypertensive emergency in a hemodialysis patient. AB - Intravenous labetolol, a nonselective alpha- and beta-blocking drug, is commonly used to treat severe hypertension. Nonselective beta-blockers can cause hyperkalemia, especially in patients with renal failure. One series reported 3 renal transplant patients who had hyperkalemia after labetolol infusion, but none of these patients developed any serious complication. We report a case of life threatening hyperkalemia (serum [K+] 9.9 mEq/l) with ventricular tachycardia and hypotension in a patient on maintenance hemodialysis who received labetolol for a hypertensive emergency. Physicians should be aware of this potentially lethal complication, which is easily preventable. PMID- 11423697 TI - Severe hypertension and massive osteoporosis as presenting symptoms of Cushing's syndrome. AB - Though Cushing's syndrome is a well-known clinical problem in terms of side effects of steroid therapy, endogenous Cushing's syndrome is a relatively rare diagnosis. We treated a 27-year-old patient who presented with severe hypertension and massive osteoporosis. We could diagnose a central Cushing syndrome by endocrinological function tests which, in retrospect, existed undiagnosed for more than 5 years. However, magnetic resonance imaging did not display an adenoma neither of the hypophysis nor of the adrenal glands. During explorative surgery, a cylindric microadenoma of the pituary gland was found and excised. After surgery, the blood pressure returned to normal, making further antihypertensive treatment unnecessary. PMID- 11423698 TI - Ultrastructure of glomerular basement membrane in active heymann nephritis rats revealed by tissue-negative staining method. AB - Recently, we have developed a tissue-negative staining method, and successfully visualized fine meshwork structure of the glomerular basement membrane (GBM). To clarify the mechanism of proteinuria in active Heymann nephritis, we performed tissue-negative staining and investigated the ultrastructural alterations of the GBM. Active Heymann nephritis, the animal model of human membranous nephropathy, was induced in Lewis rats by the injection of proximal tubular brush border antigen, i.e. Fx1A. Urinary protein excretion was measured and histological studies were performed over 15 weeks following the Fx1A injection. Proteinuria developed at 10 weeks after injection (38.2 +/- 7.4 mg/day) and progressively increased (160.2 +/- 20.6 mg/day at 15 weeks). Capillary fine deposits of IgG and C3 were seen by immunofluorescence, and subepithelial electron dense deposits (EDD) by transmission electron microscopy (TEM). Using the tissue-negative staining method, regular meshwork structure consisted of fine fibrils and pores (2.5 +/- 0.7 nm in short dimension) was observed in the GBM of control rats. At 10 and 15 weeks after injection, the GBM, directly facing the endothelial side of EDD, contained enlarged pores and nephrotic tunnels. Mean values of the short dimension of enlarged pores were 2.9 +/- 0.5 nm at 10 weeks and 3.1 +/- 0.4 nm at 15 weeks, which were significantly larger than that of control rats (p < 0.01). The rest area of the GBM, including newly produced GBM covering the epithelial side of EDD, had no significant difference in size of the pores from control GBM and no tunnels. Although there was no significant difference in the size of enlarged pores between 10 and 15 weeks, the percentage area of GBM with impaired size barrier increased at 15 weeks (51.4 +/- 8.1%) compared with 10 weeks (24.0 +/- 8.3%) and related to severity of proteinuria. The density of the tunnels also increased at 15 weeks. In conclusion, immune deposits may affect the GBM biosynthesis and induce the defect of size barrier of the GBM, which is responsible for proteinuria in active Heymann nephritis. PMID- 11423699 TI - Renal cell carcinoma in nonfunctioning transplanted kidney. PMID- 11423700 TI - Safety aspects of iron in food. AB - During the last decades efforts regarding dietary iron supply focused mostly on the prevention of deficiencies, especially during growth and pregnancy. Correspondingly, homeostatic mechanisms increase intestinal iron absorption in iron deficiency, but its downregulation at high intake levels seems insufficient to prevent accumulation of high iron stores at high intake. There is no regulated iron excretion in overload. Excess of pharmaceutical iron may cause toxicity and therapeutic doses may cause gastrointestinal side effects. Chronic iron excess, e.g. in primary and secondary hemochromatosis, may lead to hepatic fibrosis, diabetes mellitus and cardiac failure. Chronic intake of 50-100 mg Fe/day of highly bioavailable iron with home-brewed beer in sub-Saharan Africans lead to cirrhosis and diabetes. Applying a safety factor of 2 would lead to an upper safe level of 25-50 mg Fe/day for this endpoint of conventional iron toxicity. However, beyond this kind of damage iron is known to catalyze the generation of hydroxyl radicals from superoxide anions and to increase oxidative stress which, in turn, increases free iron concentration. This self-amplifying process may cause damage to lipid membranes and proteins, which relates radical generation and organ damage after ischemia-reperfusion events to available free iron in clinical and experimental settings. Correspondingly, epidemiological studies as well as observations in heterozygotes for hereditary hemochromatosis suggest that the risk of atherosclerosis and acute myocardial infarction is related to body iron stores, though there is conflicting epidemiological evidence as well. The most recent and best controlled studies, however, support the hypothesis that iron stores are related to cardiovascular risk. Iron-amplified oxidative stress may also increase DNA damage, oxidative activation of precancerogens and support tumor cell growth. This is supported by experimental, clinical and epidemiological observations. Due to these mechanisms high iron stores may present a health hazard. Though this has not been finally proven, available evidence strongly recommends not to increase iron intake beyond physiological requirements. To avoid iron deficiency symptoms, on the other hand, care must be taken to meet recommended daily intake. PMID- 11423701 TI - Testing the tyrosine/catecholamine hypothesis of oral contraceptive-induced psychological side-effects: a controlled study on triphasil. AB - BACKGROUND: This study was conducted to probe the existence of oral contraceptive (OC)-induced psychological side effects that might be undetectable by psychometric testing. METHODS: Triphasil was administered during six complete menstrual cycles to young never-OC-users. Plasma Tyr and Trp/Sigma-respective competitor neutral amino acids (NAA) ratio and concentrations of other amino acids (AA), known to be involved in the functioning of the central nervous system (CNS) and the synthesis of neurotransmitters, in particular, were used as biochemical markers to assess this likelihood. Factors known to influence plasma AA concentrations such as cortisol and pyridoxal phosphate (PLP, active form of vitamin B6), both modulators of AA intestinal absorption, were examined. RESULTS: Dietary supply of amino acid substrates (precursors) and blood levels of coenzyme/cofactor (vitamin B6 and iron) involved were adequate in both groups and were kept constant throughout the treatment. The rise in plasma cortisol, known to be stimulated under the action of estrogen, was significant (p < 0.05) after the first menstrual cycle and remained higher (although not significantly) than mean baseline and reference normal values at the end of the 6-month OC treatment. Plasma Trp/Sigma-specific competitor NAA ratio remained unchanged whereas Tyr/Sigma-specific competitor NAA ratio was significantly (p < 0.02) reduced after the sixth artificial menstrual cycle. CONCLUSION: Although plasma AA biochemical markers cannot accurately assess CNS activity, they seem to be more sensitive than a comprehensive psychometric testing (MMPI) in assessing OC induced psychological changes. PMID- 11423702 TI - Antioxidative power of plant oils in humans: the influence of alpha- and gamma tocopherol. AB - BACKGROUND: The daily diet is usually comprised of a varying antioxidant and fatty acid content acquired from plant oils. The present study was performed to evaluate the antioxidative effects of a plant oil mixture rich in alpha tocopherol (olive/sunflower oil) in comparison with a single gamma-tocopherol rich corn oil used in a normal, balanced human diet. METHODS: In the context of a double-blind study 28 healthy nonsmoking male volunteers aged between 19 and 31 years were investigated. After 2 weeks of adjustment on a mixed, balanced diet (average 11.6 MJ, average fat intake approximately 105 g/day), a 2-week test period with a diet of 80 g corn oil (CO)/day vs. a mixture of 68 g olive and 12 g sunflower oil/day (MO; total 80 g) as the main fat source, was followed by a crossover after 2 weeks. Blood samples were collected at the beginning of the study and at 2-week intervals. RESULTS: The main carriers of tocopherol in blood are low-density lipoproteins (LDLs). A highly significant correlation (r2 = 0.73, p < 0.001) was found between plasma and LDL gamma-tocopherol concentrations in the CO group, and a significant correlation was found between plasma and LDL alpha-tocopherol concentrations only in the MO group. Plasma and LDL total antioxidative capacity values could only be improved insignificantly in both groups to comparable extents, although the MO group had a twofold higher alpha tocopherol equivalent/diene equivalent ratio (1.13 mg/g) than the CO group (0.55 mg/g). CONCLUSION: In spite of the higher P/S ratio (CO 4.2 vs. MO 1.2) and lower alpha-tocopherol equivalent/diene equivalent ratio, the antioxidant potential of CO in vivo (total antioxidative capacity in plasma and LDL) was as efficient as the MO. This positive finding might be explained by a synergism between alpha- and gamma-tocopherol in the CO diet. PMID- 11423703 TI - Weight gain does not preclude increased ubiquitin conjugation in skeletal muscle: an exploratory study in tumor-bearing mice. AB - BACKGROUND AND HYPOTHESIS: At least 13 studies have shown that the ubiquitin proteasome system mediates muscle wasting in weight-losing cancer subjects. We hypothesized that cancer itself may activate the ubiquitin-proteasome system, regardless of weight loss. METHODS: We utilized hybrid mice obtained by crossing Mouse Mammary Tumor Virus-Transforming Growth Factor-alpha (TGF-alpha) mice with the Lep(ob) strain. Five hybrid MMTV-TGF-alpha heterozygous Lep(+)Lep(ob) female mice with mammary tumors were used; 4 nontransgenic heterozygous Lep(+)Lep(ob) female mice served as controls. Ubiquitin conjugates were quantitated from hamstring and paraspinal muscles by Western blotting. Myocyte apoptosis was determined by a modified TUNEL assay. RESULTS: All mice gained weight, even after tumor development. Higher concentrations of muscle ubiquitin conjugates were seen in the 5 tumor-bearing, TGF-alpha transgenic mice as compared with the 4 non tumor-bearing mice: median (range) in arbitrary densitometric units: 0.67 (0.22 4.59) versus 0.18 (0.08-0.44) in hamstring muscle and 0.56 (0.23-20.15) versus 0.18 (0.08-0.25) in paraspinal muscle (p = 0.04 and p = 0.04, respectively; Mann Whitney U test). Apoptosis was not seen in any muscle sample studied. CONCLUSIONS: Ubiquitin conjugates are increased in the skeletal muscle of tumor bearing mice in the absence of weight loss. Such activation is not seen in the skeletal muscle on non-tumor-bearing mice. Further studies might focus of whether this observation is relevant to cancer-associated wasting of lean tissue. PMID- 11423704 TI - Effect of saturated fatty acid consumption on energy and nutrient intake and blood lipid levels in preschool children. AB - In this article the effects of saturated fatty acid (SFA) consumption on energy and nutrient intake, and on a range of serum lipid parameters (indicators of cardiovascular risk) are reported in a group of 110 preschool children. Subjects were divided into 2 groups: high consumption (HC), in which > or =13% (20th percentile) of total energy came from SFAs, and low consumption (LC), in which SFAs provided <13% of energy intake. Food intake was monitored for 7 consecutive days. At participating day care centers, a 'precise individual weighing' method was used, while a 'food intake record' was kept for foods taken at home. At the blood level it was found that HC subjects showed higher HDL-cholesterol levels and better HDL-cholesterol/LDL-cholesterol ratios than did LC subjects. The greater intakes of riboflavin, iodine, zinc, magnesium and calcium seen in HC subjects suggest that these may have a better nutritive status (with respect to certain nutrients) than LC subjects. The development of criteria for providing the greatest protection against cardiovascular disease, while maintaining good nutritive condition, should be the subject of future studies. This is especially important in children of preschool age since nutritional deficiencies at this stage in life can have important effects on growth and health. PMID- 11423705 TI - The foods most consumed by German children and adolescents: results of the DONALD Study. Dortmund Nutritional and Anthropometric Longitudinally Design. AB - BACKGROUND: For a successful nutrition counseling of children and adolescents, knowledge of dietary habits is mandatory. This report describes food group intake and gives details of the customary food selection of healthy German children and adolescents. Main foods which are consumed in large amounts were identified as those on which health promotion should be concentrated. METHODS: 3-day weighed dietary records of 344 children (age range 4-6 years) and 92 adolescents (age range 13-14 years) concerning the period 1990-1997 of the DONALD Study (Dortmund Nutritional and Anthropometric Longitudinally Designed Study) were evaluated. RESULTS: The number of different foods recorded during the 3-day observation period ranged from 21 to 70 and was independent of age or sex. In contrast to dietary guidelines, the consumption of animal foods and 'fats/oils' exceeded the consumption of plant foods with the exception of adolescent girls. Food selection was very similar in age and sex groups. CONCLUSION: Our study shows that under preventive aspects the current food selection could be improved without disturbing the prevailing dietary habits and food preferences. PMID- 11423706 TI - Conflict of interest in cardiovascular publications. PMID- 11423707 TI - Platelet function and fibrinolytic agents: two sides of a coin? AB - Fibrinolytic therapy is the established treatment for the management of patients with ST elevation acute myocardial infarction (AMI). Present fibrinolytic regimens have a number of shortcomings, including the failure to produce early and sustained reperfusion, as well as failure to prevent reocclusion in at least some patients. Platelets play an important role in coronary thrombosis responsible for AMI. The effect of coronary fibrinolysis on platelets has been extensively debated in the literature with evidence of both platelet activation and inhibition. Among fibrinolytic agents, tissue plasminogen activator (t-PA) is considered to be the mainstay in the treatment of coronary artery disease. The native t-PA molecule has been modified in an attempt to achieve improved lytic characteristics with less risk of bleeding. The result is a group of mutant t-PA variants considered third-generation plasminogen activators. TNK-t-PA is one bioengineered variant of t-PA. Another third-generation plasminogen activator is reteplase (r-PA). Like TNK-t-PA, it is a variant of t-PA that has been developed to establish a more rapid, complete, and stable coronary artery patency, thus promising reduced mortality. Both r-PA and TNK-t-PA are effective when given as bolus therapy. This feature may facilitate more rapid treatment as well as decrease overall costs of treatment. New fibrinolytic regimens include potent antiplatelet agents that may improve sustained reperfusion. This review summarizes the latest and often confusing data on the interaction between fibrinolytic therapy and platelets in certain in vitro, animal and clinical scenarios. PMID- 11423708 TI - Effects of chronic nitrate therapy on left-ventricular volume in patients with heart failure secondary to coronary disease already treated with captopril: a withdrawal study. AB - OBJECTIVE: This randomized, double-blind, placebo-controlled study with treatment lasting 16 weeks and withdrawal lasting 6 weeks tried to determine whether stopping nitrates has an effect on left-ventricular end-systolic volume in patients with heart failure who were chronically treated with captopril and diuretics. PATIENTS AND METHODS: The study group comprised 29 patients with previous myocardial infarction, symptoms of mild-to-moderate heart failure, ejection fraction below 40%, no exercise-induced angina and no electrocardiographic signs of ischemia. After all patients had been treated with captopril (target dose: 25 mg twice daily), diuretics and the study drug (target dose: 40 mg isosorbide dinitrate twice daily or placebo) for 16 weeks, the study drug was withdrawn. The patients were then maintained on captopril and diuretics at constant doses for a 6-week withdrawal period. Radionuclide ventriculography with right-heart catheterization was performed at rest and during supine bicycle exercise after 16 weeks of double-blind treatment and at the end of the 6-week withdrawal period. RESULTS: The changes in resting parameters following the withdrawal of the study drug were not different between the groups. At comparable maximum workload (placebo group 68 +/- 15 W, nitrate group 68 +/- 20 W), nitrate withdrawal caused a decrease in ejection fraction (placebo withdrawal: +0.8 +/- 4.0%; nitrate withdrawal: -2.7 +/- 4.3%, p < 0.02) and increases in left ventricular end-diastolic volume (-9 +/- 35 vs. 23 +/- 48 ml, p < 0.02) and end systolic volume (-9 +/- 33 vs. +24 +/- 47 ml; p < 0.01). CONCLUSION: The addition of nitrates to a baseline therapy with captopril and diuretics might reduce exercise-induced left-ventricular dilatation in patients with heart failure from coronary disease. PMID- 11423709 TI - Cardiac troponins in suspected acute coronary syndrome: a meta-analysis of published trials. AB - We performed a meta-analysis of published trials to determine the predictive value of cardiac troponin I (cTnI) and T (cTnT) levels for adverse events (death and myocardial infarction) in acute coronary syndrome without ST elevation (ACS). The accumulated odds ratio (OR) for adverse events (30 days) in ACS with elevated cTnI (n = 5,759) and cTnT (n = 5,483) was 4.9 (95% confidence interval, CI, 3.9 6.2) and 4.6 (95% CI 3.8-5.5), respectively. Trials that mandated timed serum sampling (6 or more hours after symptom onset) had an improved predictive value for elevated cTnI (n = 2,807, OR 8.8; 95% CI 5.9-13.2) and cTnT (n = 1,990, OR 8.5; 95% CI 5.9-12.5). In conclusion, cTnI and cTnT provide similar information in ACS. The risk of adverse events is 4-fold higher in patients with suspected ACS and elevated serum cTn. For patients with an elevated timed (6-hour) sample the risk is over 8-fold higher. PMID- 11423710 TI - Plasma atrial natriuretic peptide levels for predicting the outcome of atrial fibrillation. AB - The predictive value of plasma atrial natriuretic peptide (ANP) on the cardioversion outcome was evaluated in 46 hospitalized patients with recent-onset atrial fibrillation (AF). Cardioversion was successful in 42 (91%) patients, 7 (15%) of them regained sinus rhythm spontaneously. After 12 months, 14 (33%) cardioverted patients were in chronic AF. There were no differences in plasma ANP levels between groups where cardioversion failed, those who cardioverted but later developed chronic AF or those who remained in sinus rhythm. However, among patients who were on antiarrhythmic therapy, ANP levels obtained after cardioversion were lower in those who later remained in sinus rhythm. We conclude that lower ANP after cardioversion may be associated with increased chances of long-term preservation of sinus rhythm. PMID- 11423711 TI - The influence of fetal and postnatal growth on heart rate variability in young infants. AB - Heart rate variability (HRV), a measure of cardiac autonomic control, was analyzed in infants to assess the hypothesis that early undernutrition may induce autonomic dysfunction that could play a role in the programming of later cardiovascular disease. HRV data were collected during a night session in 546 healthy infants at 5-12 weeks of adjusted age, and statistical associations with fetal and postnatal growth indices were established. A significant positive correlation between birth weight, the ratio of neonatal weight to head circumference and postnatal weight gain, and HRV indices mostly influenced by sympathetic activity was demonstrated in 11- and 12-week-old infants. A slight correlation (p > 0.05) was also found in younger infants. These data suggest the influence of fetal and postnatal growth on the programming of the autonomic nervous system beyond the neonatal period. This influence may be one of the important mechanisms that link impaired growth in fetal and infant life to high blood pressure and other cardiovascular disease during childhood and adulthood (the Barker hypothesis). PMID- 11423712 TI - Tissue factor expression in coronary circulation as a prognostic factor for late restenosis after coronary angioplasty. AB - We investigated changes in blood coagulation in the coronary circulation after percutaneous transluminal coronary angioplasty (PTCA) and its clinical significance. We examined 43 patients with ischemic heart disease who underwent elective PTCA of isolated stenotic lesions in the left coronary artery. Ten patients underwent PTCA alone, 15 received percutaneous transluminal rotational atherectomy (PTRA) and 18 stent implantation. Blood samples were drawn from the coronary sinus before and immediately after PTCA, as well as 4 and 24 h later. Plasma levels of tissue factor (TF), thrombin-antithrombin III complex (TAT) and prothrombin fragment 1+2 (F 1+2) were measured by enzyme-linked immunosorbent assay. Follow-up coronary angiography was performed 6 months after PTCA. Minimal luminal diameter was assessed by quantitative coronary angiography to evaluate late loss index. TF, TAT and F 1+2 levels in the coronary sinus blood showed significant increases 24 h after PTCA. A significant positive correlation was found between changes in TF levels 24 h after PTCA and late loss index 6 months after the procedure. TF levels in the coronary sinus blood were significantly higher in patients with late restenosis than in those without restenosis. These results suggest that TF expression in the coronary circulation after PTCA is a prognostic factor for late restenosis. PMID- 11423713 TI - Influence of the availability of laser transmyocardial revascularisation on surgical strategy in patients with advanced coronary artery disease. AB - OBJECTIVE: To compare the planned and actual revascularisation techniques used in patients undergoing cardiac surgery for refractory angina when excimer transmyocardial laser revascularisation (TMR) is available. METHODS: Observational series of 31 patients (mean age 65 years) with severe angina [mean CCS score (SD) 3.8 (0.4)] and three-vessel coronary artery disease judged unsuitable for conventional bypass surgery alone. All patients underwent cardiac surgery: revascularisation techniques were determined by the operative findings. RESULTS: Pre-operative strategy was altered by the intra-operative findings in 13 patients (42%). In 5 (16%), the coronary vessels proved graftable and TMR was unnecessary. Conversely, in 6 patients (19%) an anticipated graft could not be performed and TMR was used as an alternative. In 2 patients (7%), neither strategy was possible. Overall, TMR was performed as a stand-alone procedure in 9 (29%) and combined with CABG in 17 (55%). Operative mortality was low: 0% at 30 days and 6% at 6 months. Mean CCS class (SD) improved post-operatively from 3.8 (0.4) to 1.7 (1.1) (p < 0.01). CONCLUSIONS: The pre-operative coronary angiogram is an imperfect predictor of which coronary vessels are suitable for grafting. The availability of laser TMR allows the cardiac surgeon to accept cases which would otherwise be considered inoperable and to respond better to intraoperative findings. The combination of laser TMR and bypass grafts provides good short- and medium-term symptomatic improvement with a low post-operative mortality. PMID- 11423714 TI - Echocardiographic evaluation of left-ventricular diastolic function in patients with chronic pulmonary hypertension. AB - Different patterns of left-ventricular (LV) diastolic dysfunction were reported in patients with pulmonary hypertension (PHT). There are no data regarding the relationship between the severity of PHT and LV diastolic dysfunction. In order to determine the severity of PHT at which LV diastolic dysfunction occurs and to identify its pattern, we studied by Doppler echocardiography 120 patients with PHT (57 with severe PHT and 63 with mild or moderate PHT) and compared them with 75 normal controls. Systolic pulmonary artery pressure (SPAP) was measured by tricuspid regurgitant jet method and the usual transmitral LV diastolic indices were recorded. LV diastolic dysfunction of impaired relaxation type is most commonly seen in patients with severe PHT. No differences were observed between patients with mild and moderate PHT regarding LV diastolic function. A SPAP > or =60 mm Hg is needed to induce changes in the LV diastolic filling pattern. PMID- 11423715 TI - Interrogation of the tricuspid annulus by Doppler tissue imaging in patients with chronic pulmonary hypertension: implications for the assessment of right ventricular systolic and diastolic function. AB - BACKGROUND: Chronic pulmonary hypertension (CPHT) has a great impact on both right-ventricular (RV) systolic and diastolic properties and the assessment of those properties is not always feasible by traditional echocardiographic examination. Doppler tissue imaging (DTI) interrogation of the tricuspid annulus (TA) identifies the pattern of TA motion (TAM) and can help to assess RV function when other methods are not feasible. AIMS: To determine RV systolic and diastolic function in patients with CPHT using DTI parameters of the TA. METHODS: Eighty seven patients with CPHT and 90 normal controls were studied. DTI parameters were measured including early diastolic, late diastolic and systolic velocities and time velocity integrals (TVI) of the TAM at both its lateral and medial aspect. RESULTS: Early diastolic and systolic velocities, the ratio of early to late diastolic velocities and TVI of TAM at both lateral and medial aspects were significantly decreased in patients with CPHT compared to controls. No significant differences were seen in late diastolic velocities and TVI in both groups. Systolic velocity of the TAM at both its lateral and medial aspects significantly correlated with RV systolic function as measured by fractional RV area change. CONCLUSIONS: DTI of the TAM can be used to assess RV systolic and diastolic properties in patients with CPHT. PMID- 11423716 TI - The Doppler echocardiographic myocardial performance index predicts left ventricular dilation and cardiac death after myocardial infarction. AB - To investigate the value of the Doppler-derived myocardial performance index to predict early left-ventricular (LV) dilation and cardiac death after a first acute myocardial infarction (AMI), Doppler echocardiography was performed within 24 h of hospital admission, on day 5, 1 and 3 months after AMI in 125 consecutive patients. The index measured on day 1 correlated well with the change in end diastolic volume index observed from day 1 to 3 months following AMI (r = 0.66, p < 0.0001). One-year survival in patients with Doppler index <0.63 was 89%, and 37% in patients with index > or = 0.63, (p < 0.0001). Multivariate analysis identified myocardial performance index > or = 0.63 (relative risk 5.6, p < 0.0001), E-wave deceleration time <140 ms (relative risk 2.7, p = 0.008) and Killip class > or = II (relative risk 4.0, p = 0.04) to be independent predictors of cardiac death. Therefore, we conclude that the Doppler echocardiographic myocardial performance index is a predictor of LV dilation and cardiac death after a first AMI. PMID- 11423717 TI - Comparison of adenosine and exercise stress test for quantitative perfusion imaging in patients on beta-blocker therapy. AB - Beta-blocker therapy is used to decrease myocardial ischemia during exercise but may cause suboptimal diagnostic performance in exercise stress testing. The aim of the present study was to compare results of quantitative technetium-99 sestamibi single photon emission tomography (SPECT), following exercise stress test or pharmacological stress test with adenosine. We chose adenosine as comparison, since betablockers may not interfere with adenosine induced vasodilatation and therefore possibly may not interfere with its diagnostic performance. Sixteen patients with angiographically documented coronary disease (5 single-vessel, 6 two-vessel and 5 three-vessel disease), who were chronically treated with beta-blockers, performed SPECT imaging at rest, following bicycle exercise and following adenosine infusion in random order. The SPECT data were analyzed visually and quantitatively, using dedicated computer software (CEqual). According to both visual and quantitative SPECT analysis, adenosine was superior to show reversibility. Higher reversibility extent (50 +/- 15 vs. 26 +/- 12 pixels, p < 0.01) and more intense reversibility severity (110 +/- 29 vs. 49 +/- 23 sum of SDs, p < 0.05) were observed during adenosine than exercise. CONCLUSIONS: Less myocardial perfusion abnormalities during exercise than during adenosine stress in patients treated with beta-blockers may indicate less ischemia but also an impaired diagnostic performance. Thus adenosine stress test should be preferred to optimize the diagnostic sensitivity in patients during beta-blocker treatment. PMID- 11423718 TI - The association between asthma and dental caries in children and adolescents: A population-based case-control study. AB - This study explored the potential association between childhood asthma and caries using oral examination and health interview data from the Third National Health and Nutrition Examination Survey 1988-1994 (NHANES III). We fitted GEE Poisson regression models with adjustment for parents' income, gender, race, exposure to potentially xerostomic drugs, and the presence of pit and fissure sealants. There was no association between the use of drugs commonly used by asthmatics (antihistamines, corticosteroids, and antiasthmatic inhalers) and df/DMF scores. Asthmatic children 4-10 years of age at all severity levels had similar dfs scores to the controls, however, severely asthmatic children 4-10 years of age had significantly lower DMFS (p = 0.010) and DMFT (0.049) scores than controls. Similarly, severely asthmatic children 11-16 years of age had significantly lower DMFT scores than controls (p = 0.024) and DMFS scores approaching statistical significance (p = 0.053). While our analysis adjusted for covariates, potential confounders such as fluoride intake from water, diet, use of topical fluorides, and dose of antiasthmatic medication could not be addressed. Our results indicate that any association between asthma and dental caries may occur primarily in younger children with no evidence of an association between asthma and dental caries as children mature. PMID- 11423719 TI - The change of DMFT counts in Slovenia. AB - This longitudinal study was designed to assess the change in caries prevalence and DMFT counts in the adult population of Ljubljana, Slovenia, over a 10-year period. All data in the present study are derived from two surveys conducted in 1983-1987 and 1993-1997. In the first survey, 1,609 dentate subjects in the range from 15 to 65 years were examined. Ten years later, 555 (34.5%) of these subjects participated in the second oral survey. In the 538 dentate 25- to 75-year-old subjects examined in the second survey, a decline in the mean number of decayed teeth (1.1 vs. 0.5) was found, and an increase in DMFT counts (15.9 vs. 19.1) due to a higher number of missing (5.2 vs. 7.5) and filled teeth (9.7 vs. 11.1) was observed. However, a comparison with the previous generation of the same age showed a decline in the average number of decayed and missing teeth in all age groups and an increase in the mean number of filled teeth in all age groups as well. The population of Ljubljana from the 1993-1997 survey had retained in most of the age groups about 2 teeth more than had the generation 10 years before. PMID- 11423720 TI - Determination of fluoride intake from urinary fluoride excretion data in children drinking fluoridated school milk. AB - Twenty-four-hour urine samples were collected from each of thirteen 5- to 6-year old children both under customary conditions of fluoride intake (i.e. usual diet including milk containing 0.5 mg fluoride and usual oral hygiene procedures with fluoride toothpaste) and during a 4-day study period in which their customary fluoride intake was replaced with standard fluoride doses. The 24-hour fluoride excretion under customary conditions of fluoride intake was 0.30 mg and in response to fluoride doses in the range of 0.5-2.0 mg it was in the range 0.26 0.61 mg. Using data obtained during the study period for all subjects combined, the fluoride intake from all sources under customary conditions was estimated as 0.76 mg, representing a fractional fluoride excretion of 39%. It is concluded that these children are possibly receiving suboptimal amounts of fluoride. PMID- 11423721 TI - Peroxidase reaction as a parameter for discrimination of Streptococcus mutans and Streptococcus sobrinus. AB - 425 strains of mutans streptococci and 12 reference strains were investigated by membrane fatty acid spectra (MFAS) and peroxidase reaction (PR) after aerobic and anaerobic incubation. 423 strains were identified as Streptococcus mutans. The remaining 2 strains were identified as Streptococcus sobrinus. The PR of 29 strains was doubtful; immediately after anaerobic incubation a negative PR changed into a slightly positive PR. To test the diagnostic value of PR the strains were additionally investigated by means of species-specific polymerase chain reactions (PCR). The species-specific PCRs were developed on the basis of the respective genes of 16S rRNA of the pathogens S. mutans and S. sobrinus. Specificity and sensitivity were tested on reference strains (n = 17) and negative control strains (n = 39). The results of this investigation showed that an anaerobic incubation regime could lead to false-positive (S. mutans) or false negative (S. sobrinus) PR. The 425 MS strains were classified as either S. mutans (n = 420) or S. sobrinus (n = 5). The findings on the reference strains required a reclassification of S. mutans V 100 into S. sobrinus V 100. Summarising, it is possible now to differentiate strains of mutans streptococci by MFAS and PR after aerobic incubation. PMID- 11423722 TI - Insights into the role of the von Hippel-Lindau gene product. A key player in hypoxic regulation. AB - Many adaptive responses to hypoxia involve changes in gene transcription mediated by the hypoxia-inducible factor 1 complex. Central to this is oxygen-dependent proteolysis of the alpha subunit, which has recently been shown to require the von Hippel-Lindau tumour-suppressor protein. This observation provides one mechanism by which inherited defects in the von Hippel-Lindau gene could cause features of the clinical syndrome, and offers insight into the events leading to sporadic clear cell renal cancer. Furthermore, it clearly implicates the von Hippel-Lindau tumour-suppressor protein in the biochemistry of oxygen sensing. PMID- 11423723 TI - Chemokine expression in the obstructed kidney. AB - Chemokines are chemotactic cytokines that are important mediators of leukocyte extravasation and chemotaxis. Herein, we provide evidence that after 1 day of unilateral ureteral obstruction (UUO), the mouse obstructed kidney (OBK) expresses MCP-1 (monocyte chemoattractant protein-1), RANTES (Regulated on activation normal T-cell expressed and secreted) and IP-10 (interferon-gamma induced protein-10). In addition, by day 7, MIP-2 (macrophage inflammatory protein-2) expression is elevated in the obstructed kidneys compared to the contralateral control kidneys (CLK). After 7 days of obstruction, RANTES was the most abundant of the four chemokines detected in the OBK. In situ hybridization results indicate that several cellular compartments contribute to the expression of RANTES in the OBK. However, clearly cortical tubules within the OBK contribute substantially to the elevated expression of RANTES. These data support the contention that the cortical tubular epithelium plays a pivotal role in the inflammation associated with experimental hydronephrosis. PMID- 11423724 TI - High glucose decreases matrix metalloproteinase-2 activity in rat mesangial cells via transforming growth factor-beta1. AB - Diabetic nephropathy is characterized by accumulation of mesangial matrix. Glucose-induced inhibition of matrix-degrading enzymes such as collagenases is believed to contribute to matrix accumulation. We have previously demonstrated that 72 kDa type IV collagenase activity is decreased in the rat mesangial cells cultured in high glucose media [Diabetes 1995;44:929-935]. The present studies were designed to investigate if the cytokine transforming growth factor-beta1 (TGF-beta1) mediates this effect of glucose. Type IV collagenases degrade type IV collagen as well as gelatin (denatured collagen) and are thus also called gelatinases. They belong to the family of matrix metalloproteinases (MMPs); MMP activity is controlled by tissue inhibitors of metalloproteinases (TIMPs). The activity of 72 kDa type IV collagenase, also known as matrix metalloproteinase-2 (MMP-2), was assessed using three methods: (1) fluoresceinated gelatin degradation assay to detect free enzyme activity (activity which is present in excess of TIMP-inhibited activity); (2) zymography to measure total (free + TIMP bound) enzyme activity; (3) ELISA using specific antibodies to measure MMP-2 levels. TGF-beta1 and TIMP-2 levels were also determined by ELISA. Incubation of primary cultures of rat mesangial cells for 5 days in 30 vs. 5 mM glucose resulted in a 3-fold increase in production of total TGF-beta1, a significant decrease in MMP-2 activity and immunoreactive MMP-2 levels, and an increase in TIMP-2 levels. Addition of exogenous TGF-beta1 to mesangial cells incubated in 5 mM glucose replicated the high glucose effect by producing a significant decrease in MMP-2 levels with a concurrent increase in TIMP-2 levels. Furthermore, glucose induced inhibition of MMP-2 activity was completely blocked by neutralization of TGF-beta1 with anti-TGF-beta1 antibody. We conclude that the decrease in MMP-2 activity induced by glucose loading is mediated via TGF-beta1. PMID- 11423725 TI - Phosphate depletion in opossum kidney cells: apical but not basolateral or transepithelial adaptions of Pi transport. AB - Monolayers of opossum kidney (OK) cells are widely used as models for the renal proximal tubule. OK cells adapt to phosphate (Pi) depletion by increasing their capacity for apical and basolateral Na+-dependent Pi uptake. Because NMR-visible cell Pi was found to be decreased in Pi-deprived kidney cells, we suggested that up-regulation of basolateral Pi efflux also occurs during adaptation to Pi deprivation [American Journal of Physiol 1994;267:C915-919]. In order to test this hypothesis, we measured the cell Pi pool, basolateral Pi efflux and transepithelial Pi fluxes in OK cells grown on permeable plastic filters, exposed overnight to solutions containing either 0.5 mM (deprived) or 2.0 mM (replete) Pi or 32Pi. Following steady state or acute loading with 32Pi, the specific activity (SA) of cell Pi, the cell Pi pool and the basolateral efflux of 32Pi were measured. In the steady state, a 2-fold increase in Pi uptake sustained the intracellular Pi pool at 85% of the control level (30 +/- 5 nmol/mg) in spite of a decrease in extracellular Pi from 2 to 0.5 mM. When the extracellular Pi was acutely (1 h) reduced to 0.1 mM, the cell Pi pool decreased (to 3 +/- 1 nmol/mg) both in cells previously adapted overnight to either 0.5 or to 2 mM Pi (p >0.3). The rates of absolute and fractional basolateral washout of cell 32Pi after 1 h loading with 0.1 mM 32Pi were similar in cells adapted to 0.5 compared to 2 mM Pi. This indicates that Pi depletion did not affect the effective permeability of the basolateral membranes to Pi. Adaptation for 16 h to 0.5 compared to 2 mM Pi did not alter the rate of net transepithelial transport of 0.1 mM Pi from the apical to the basal compartment but reduced (p < 0.05) the unidirectional fluxes of both 32Pi and 14C-mannitol. An insufficient driving force (unchanged or low Pi concentration in the transport pool, low electrical or coupled-anion gradients) and a constant effective basolateral Pi permeability must have limited basolateral Pi efflux in cells exposed to 0.1 mM Pi. Thus, in OK cells grown on plastic support there are no adaptive increases in either basolateral Pi efflux, or in transcellular and paracellular Pi transport, in response to Pi depletion. Adaptations are limited to increases in apical and basolateral sodium-dependent Pi uptakes that can maintain the cell Pi pool as long as apical Pi is not too low (> or =0.5 mM). The OK cells adapt to low Pi concentrations conserving cell Pi but not increasing basolateral Pi efflux nor transepithelial Pi transport. PMID- 11423726 TI - Molecular cloning and characterization of profilin-3: a novel cytoskeleton associated gene expressed in rat kidney and testes. AB - Profilin is a small actin-binding protein that is involved in diverse functions such as maintaining cell structure integrity, cell mobility, tumor cell metastasis, as well as growth factor signal transduction. In this paper, we describe the molecular cloning and characterization of a novel form of profilin, termed profilin-3, from rat kidney using a PCR-based procedure for isolating tissue-specific genes. The profilin-3 cDNA encoded 137 amino acids, and it shared extensive homology to profilin-1 and profilin-2 from mice and humans. More strikingly, the expression of profilin-3 was highly selective, and its mRNA was only found in the kidney and to a much lesser extent in the testis. The size of the mRNA for profilin-3 in the testis was 1.2 kb, while in kidney it was approximately 4.4-5 kb, suggesting the presence of tissue-specific transcription from different promoters. In addition, we also found that the expression of profilin-3 mRNA was significantly elevated in two types of renal diseases: diabetic nephropathy (db/db mice) and polycystic kidney diseases (cpk mice). Similar to profilin-1 and profilin-2, profilin-3 was localized in the cytoplasmic domain. Furthermore, it was capable of interacting with actin and poly-L-proline in an in vitro assay as well as in transfected cells. These results strongly suggest that, contrary to the ubiquitous presence of profilin-1 and profilin-2, there is a tissue-specific form of these cytoskeleton-regulatory proteins and that the kidney/testes-specific profilin-3 may play a unique role in renal and/or reproductive functions. PMID- 11423727 TI - Effects of type 2 cytokines on glomerular epithelial cells. AB - Visceral glomerular epithelial cells (GECs) are involved in the maintenance of the filtration barrier and may play a role in immune responses. Cytokines may act on GECs and we wished to test this in vitro. Vascular endothelial growth factor (VEGF) is a specific product of the GEC that may play a role in glomerular permeability. We have investigated whether GECs in culture express receptors for interleukin (IL)-4, 10 and 13 (often grouped together as type 2 cytokines) and whether these cytokines alter GEC VEGF production. Type 2 cytokines were compared to transforming growth factor-beta (TGF-beta) and IL-1beta which are known to upregulate VEGF production. GECs were grown from human nephrectomy specimens and cultured with and without the addition of exogenous cytokines. Messenger RNA data demonstrated the presence of IL-4 receptor alpha, IL-10 receptor 1 and 2, and IL 13 receptors alpha1 and alpha2. However, at the protein level by flow cytometry, only IL-13 alpha2 could be consistently demonstrated. IL-4, IL-10 and IL-13 inhibited production of VEGF but did not affect the pattern of isoform expression. In contrast, TBF-beta and IL-1beta caused an increase in VEGF production. These effects were not explained by effects on proliferation. Our data provide evidence that GECs express receptors for type 2 cytokines and that these cytokines can act directly on GECs, to decrease VEGF production. PMID- 11423728 TI - Preconditioning with sodium arsenite inhibits apoptotic cell death in rat kidney with ischemia/reperfusion or cyclosporine-induced Injuries. The possible role of heat-shock protein 70 as a mediator of ischemic tolerance. AB - This study was performed to evaluate the effect of heat-shock protein (HSP)70 induction with sodium arsenite (SA) on ischemia/reperfusion (I/R) or cyclosporin A (CsA)-induced injuries in rat kidney. Rats were classified into five groups (sham, I/R, SA+I/R, I/R+CsA and SA+I/R+CsA groups) according to both the status of SA pretreatment and treatment with CsA. SA (6 mg/kg, i.v.) pretreatment was accomplished 12 h before I/R injury, and CsA (20 mg/kg, s.c.) was given subsequent to I/R injury. The effect of SA pretreatment on I/R injury was evaluated using measurements of renal function, the histopathology score, and assays for apoptosis (DNA fragmentation analysis, TUNEL staining, mRNA expressions of the pro-apoptotic genes and caspase activities). In addition, mitochondrial morphology was examined by electron microscopy. Induction of HSP70 with SA improved both renal function and the histopathology score as compared to the group without HSP70 induction. The assays for apoptosis revealed that SA pretreatment decreased the DNA laddering pattern, TUNEL-positive cells, mRNAs expression of pro-apoptotic genes and caspase activities as compared with the group without SA pretreatment. In addition, the mitochondrial morphology was well preserved in the groups with SA pretreatment. In conclusion, SA pretreatment prevents subsequent I/R or CsA-induced injuries in the rat kidney, and this renoprotective effect appears to be mediated by induction of HSP70. PMID- 11423729 TI - [Gynakologisch-geburtshilfliche Rundschau -revisited, future directed and practice oriented]. AB - After a long period of 15 years, the SGGG recognized the Gynakologisch geburtshilfliche Rundschau once again as their official organ. Herewith a sign is given for a more intense cooperation with the Austrian Society for Gynecology and Obstetrics. Eventually, the possibility that the Bavarian Society of Gynecology and Obstetrics will also join might be achieved. In a time of rapid developments in the field of print media, publishers and editors are now facing new and upcoming challenges. The concept of our journal will be adapted to the modern requirements of our readership. Besides the traditional original papers, important with regard to the impact factor, other topics will be published, e.g., medicine and law, management and health care, and congress reports. As recently introduced, articles in German as well as in English are welcome. PMID- 11423730 TI - [Roberts-SC phocomelia syndrome]. AB - The Roberts-SC phocomelia syndrome is a rare autosomal recessive inherited disorder clinically manifested by tetraphocomelia, pre- and postnatal growth retardation, and craniofacial abnormalities (skull, eyes, lip, and palate), accompanied at times by centromer puffing and splitting, renal abnormalities, heart defect, clitoral or penile enlargement, and bilateral corneal opacities. Mental retardation is common in surviving patients. PMID- 11423731 TI - [HELLP Syndrome as an indication for liver transplantation]. AB - Liver rupture followed by multiorgan failure is perhaps the most catastrophic complication of the HELLP syndrome (hemolysis, elevated liver function, low platelets). Specific treatment options are currently limited and not routinely clarified. Here, we present a patient successfully managed by an innovative surgical approach consisting of combined total hepatectomy, portacaval shunt, and liver transplantation. A 26-year-old primipara (39th week of gestation) who suffered liver rupture as a complication of HELLP syndrome after delivery underwent a portacaval shunt after total hepatectomy. This combination was sufficient until the patient underwent orthotopic liver transplantation. The patient was discharged from the hospital after a dramatic recovery. Bridging portacaval shunt and consecutive orthotopic liver transplantation represented an effective therapy for this patient and should be considered early as a treatment option in patients with liver rupture complicating severe HELLP syndrome. PMID- 11423732 TI - [Recommendations for replacement of estrogens and progestins in climacteric and in postmenopause. 25. Arbeitstreffen des "Zurcher Gesprachskreises", November 2000]. PMID- 11423733 TI - [Critical appraisal of medical literature:pragmatic approach]. AB - Due to the enormous amount of medical information in journals and the Internet, keeping doctors up to date has become increasingly difficult. A pragmatic approach to reduce the amount of information is to select on the basis of the topic, the title, and the abstract. After this selection, reading should not be limited to the abstract alone. Abstracts go short on essential information about the applicability of the information in clinical practice. To read the whole article might be time-consuming, but the validity of the data has to be evaluated prior to application to patients. PMID- 11423735 TI - Importance of early postnatal germ cell maturation for fertility of cryptorchid males. AB - PATIENTS/METHOD: Twenty-seven adults who underwent an orchidopexy before 2 years of age have recently had their spermiograms analyzed. RESULTS: Sixty-three percent of them had a normal sperm count; their germ cells had undergone the second stage of maturation and adult dark (Ad) spermatogonia were present at the time of surgery 20-25 years earlier. In contrast, 37% of them were infertile despite a seemingly successful orchidopexy; they had impaired second-step germ cell maturation and Ad spermatogonia were absent at orchidopexy. CONCLUSIONS: Transformation of gonocytes into Ad spermatogonia is crucial for fertility. This underscores the importance of a testicular biopsy at orchidopexy in order to identify those who could benefit from early LH-RH treatment after orchidopexy. PMID- 11423736 TI - Abnormal germ cell development in cryptorchidism. AB - BACKGROUND: Previous studies suggest that two fundamental, probably androgen dependent, steps in maturation of germ cells normally occur in the prepubertal testis: the disappearance of gonocytes (the fetal stem cell pool) and the appearance of adult dark spermatogonia (the adult stem cell pool) at 2-3 months of age and the appearance of primary spermatocytes (the onset of meiosis) at 4-5 years. Previous studies of small series of cryptorchid boys suggest that both steps are defective in undescended testes and to a lesser degree in descended testes contralateral to unilaterally undescended testes. The purpose of this study is to confirm the previous findings of defective germ cell maturation in a large series of boys with unilateral undescended testes. PATIENTS: Seven hundred and sixty-seven boys with unilateral cryptorchidism who had orchidopexy and bilateral testicular biopsies between birth and 9 years of age were studied. MATERIALS AND METHODS: Total and differential germ cell counts were performed on semithin histologic sections of the biopsies. The results from the undescended and contralateral descended testes were compared using the Wilcoxon signed-rank test and the Wilcoxon-Whitney-Mann U test. RESULTS: Gonocytes failed to disappear and adult dark spermatogonia failed to appear in undescended testes under 1 year of age indicating a defect in the first step in maturation at 2-3 months resulting in failure to establish an adequate adult stem cell pool. Primary spermatocytes failed to appear in undescended testes and appeared in only 19% of contralateral descended testes at 4-5 years of age indicating a defect in the onset of meiosis. CONCLUSION: Unilaterally undescended testes fail to establish an adequate adult stem cell pool which normally occurs at 2-3 months of age and fail to establish adequate meiosis which normally occurs at 4-5 years of age. Similar but less severe changes are seen in the contralateral descended testes. Defects in the two pubertal steps in germ cell maturation are associated with reduced total germ cell counts. PMID- 11423737 TI - Fertility potential: a comparison of intra-abdominal and intracanalicular testes by age groups in children. AB - We compared a group of intra-abdominal (IA) with intracanalicular (IC) testes of the same age in order to see if there were differences in the fertility potential of the testes in these two locations. We also wished to determine at what age, if any, differences became apparent. Semithin (1 microm thick) sections of 54 IA and 103 IC testes between the ages of 3 months and 5 years were evaluated and the spermatic index (SI) calculated for each. The two groups were compared using mathematical models to describe the relationship between age and SI. Models of both IA and IC testes show a sharp decline in the SI during the first 1.8 years of life and remain near zero thereafter. The models are statistically indistinguishable, suggesting there is no detectable difference in the fertility potential of IA and IC testes up to 5 years. The SI appears to reach a critically low value in both groups between 8 and 9 months of age, suggesting that surgical intervention would be appropriate before this time. PMID- 11423738 TI - Cryptorchidism: aspects of fertility and neoplasms. A study including data of 1,335 consecutive boys who underwent testicular biopsy simultaneously with surgery for cryptorchidism. AB - PURPOSE: An attempt to make a rational strategy for treatment of cryptorchidism. MATERIALS AND METHODS: 1,335 cryptorchid boys with biopsy at surgery (1,638 specimens). We studied: frequency of no germ cells in biopsies from 698 patients <12 years at surgery; fertility potential of 140 patients who were now adults, and apperance of testicular neoplasia in all biopsies. RESULTS: Lack of germ cells appeared from 18 months. The frequency increased with increasing age. It appeared in 30% (61/202) bilateral, and 18% (88/496) unilateral cases. In men who had undergone bilateral or unilateral orchiopexy, respectively, there was normal sperm count in 19% (14/75) and 83% (54/65), and infertility was suspected in 56% (42/75) and 8% (5/65) (FE, p < 0.00005, p < 0.00005), respectively. The lowest, the mean, and the highest age-matched spermatogonia count per tubule at orchiopexy was associated with sperm count (Spearman test, p < 0.0001, p < 0.005, p < 0.05). Isolated, this was demonstrated for the 75 formerly bilateral (Spearman, p < 0.0001, p < 0.0001, p < 0.0001), but not the 65 formerly unilateral cases (Spearman, p = 1.0). No germ cells at orchiopexy was associated with suspected infertility. Risk was 78-100% in bilateral (dependent on one or both testes affected), and 33% in unilateral cryptorchidism. There was one invasive germ cell tumor, six cases of carcinoma in situ testis, and one Sertoli cell tumor. Three neoplasms were diagnosed in intra-abdominal testes, four in boys with abnormal external genitalia, and two in boys with known abnormal karyotype. Risk of neoplasia was 5% (7/150) in patients with intra-abdominal testis, abnormal external genitalia or diagnosed abnormal karyotype, versus 0% (0/1,185) in patients without these characteristics (FE, p < 0.00005). CONCLUSION: We recommend surgery for cryptorchidism before 15-18 months of age because: (a) lack of germ cells is very rare before, and (b) lack of germ cells is associated with subsequent risk of infertility. At primary surgery for cryptorchidism, we recommend examination for testicular neoplasia in cases of intra-abdominal testis, abnormal external genitalia or known abnormal karyotype. PMID- 11423739 TI - Fertility after bilateral cryptorchidism. Evaluation by paternity, hormone, and semen data. AB - PURPOSE: Evaluation of the fertility of a cohort of formerly bilaterally cryptorchid men in comparison with a group of formerly unilaterally cryptorchid men, and a group of control men. MATERIALS AND METHODS: Using a detailed questionnaire concerning paternity and factors related to paternity, a cohort of formerly bilateral cryptorchid men were studied and compared with men who had undergone orchiopexy for unilateral cryptorchidism, and a group of control men. All study subjects had had surgery at the Children's Hospital of Pittsburgh, Pittsburgh, Pa., between 1955 and 1975. A subset of the full cohort underwent clinical evaluation that included a physical examination, serum hormonal determination and semen analyses. RESULTS: Paternity rates are significantly lower among the formerly bilaterally cryptorchid men who have attempted to father a child (65.3%) as compared to the formerly unilaterally cryptorchid (89.7%; p < 0.001) and control men (93.2%; p < 0.001). Differences in the ability to father children are also apparent when semen and hormone levels are compared between the three groups. The bilateral group has significantly lower sperm density and inhibin B levels, and higher FSH and LH levels, than the unilateral and control groups. CONCLUSIONS: Men born with bilateral cryptorchidism have severely compromised fertility in adulthood. This reduction in fertility is clearly shown in comparisons of both paternity rates, and in semen and hormone analyses, between the formerly bilateral, formerly unilateral, and control groups. PMID- 11423740 TI - 13 Years' experience with the combined hormonal therapy of cryptorchidism. AB - PURPOSE: We analyze the results of the combined treatment with luteinizing hormone releasing hormone (LH-RH) and human chorionic gonadotropin (HCG) of a large series of patients with cryptorchidism. MATERIALS AND METHODS: Between 1987 and 1999 and after strict differentiation between cryptorchid, retractile and gliding testes, 2,467 boys with 2,962 cryptorchid-gliding testes were treated with the combined hormonal therapy. LH-RH was administrated as a nasal spray at a dosage of 1.2 microg daily for a period of 4 weeks. HCG was injected intramuscularly, 5 times at 2-day intervals at a dosage adjusted according to the age. RESULTS: In the prospective study 2,476 boys with 2,962 cryptorchid testes were hormonally treated. Of the 2,962 evaluated cases 1,200 (40.52%) have been treated surgically after the hormone therapy. In 1,762 cases, 59.48% of cryptorchid testes were in the scrotum after combined hormone treatment. CONCLUSIONS: Treatment with LH-RH and HCG induced the descent of the testes to a normal scrotal position of boys with cryptorchidism in 59.48% of the evaluated cases. The combined treatment was effective for inducing descent of cryptorchid and gliding testes. According to the evaluated intraoperative findings, the failure of the combined therapy in 40.52% of the cases is due to the fact that the free descent is limited by local factors such as anatomical alterations of the inguinal canal, epididymal abnormalities or ectopic distal attachment of the lig. testis. PMID- 11423741 TI - Hormonal therapy for the subfertility of cryptorchidism. AB - BACKGROUND: The subfertility of cryptorchidism correlates with severely reduced total germ cell counts in prepubertal testicular biopsies of undescended testes. Reduced total germ cell counts are associated with defects in the two prepubertal steps in maturation and proliferation in germ cells: first, the transformation of the fetal stem cell pool (gonocytes) into the adult stem cell pool (adult dark spermatogonia) at two to three months of age and, second, the transformation of adult dark spermatogonia into primary spermatocytes (meiosis) at 4-5 years. The defects in maturation are associated with blunting of the normal surges in gonadotropins and testosterone. Prepubertal treatment with gonadotropin-releasing hormones would theoretically trigger normal germ cell maturation and proliferation and thereby improve total germ cell counts and improve fertility. Prepubertal treatment of cryptorchidism with the GnRH analogue Buserelin has resulted in improved total germ cell counts and improved spermiograms. The purpose of this report is to describe the results of treatment of cryptorchidism with the GnRH analogue Naferelin. PATIENTS: Twelve boys with cryptorchidism, 6 unilateral and 6 bilateral, and severely reduced germ cell counts in testicular biopsies were treated with Naferelin following successful orchidopexy and bilateral testicular biopsies. Response of the total germ cell counts was assessed in follow-up bilateral biopsies within 5 months of completing the hormonal therapy. RESULTS: Eight of the 12 boys (5 of the 6 with unilateral and 3 of the 6 with bilateral cryptorchidism) showed improvement in the total germ cell counts in one or both testes. All 8 had a poor prognosis for fertility pretreatment and a good prognosis for fertility posttreatment. Of the 5 with unilateral cryptorchidism who improved, 2 showed improvement in both testes; and 3, only in the contralateral descended testes. All 3 of the boys with bilateral cryptorchidism who improved showed improvement in both testes. Testes with absence of germ cells and older patients tended to show no improvement. Of the 6 contralateral descended, 5 (83%) improved, and of the 18 undescended testes, 8 (44%) improved. CONCLUSIONS: In this preliminary study, Naferelin therapy appears to induce improvement in the total germ cell counts and the prognosis for future fertility in 75% of patients. PMID- 11423742 TI - Erythropoietin may reduce the risk of germ cell loss in boys with cryptorchidism. AB - PURPOSE: In boys with cryptorchidism older than 2 years a testicular biopsy at time of orchiopexy shows lack of germ cells in 10-40% of the cases. The number of spermatogonia per tubule is prognostic for subsequent fertility potential. A biopsy without germ cells is associated with 33-100% risk of infertility. In order to increase the number of germ cells, and thereby the fertility potential, additional hormonal therapy has been attempted before surgery. In a study, small doses of the gonadotropin-releasing hormone analogue buserelin before orchiopexy caused higher values. Others have found that hormonal treatment with human chorionic gonadotropin or gonadotropin releasing hormone analogue may harm the germ cells in cryptorchidism. The aim of the study is to demonstrate that additional hormonal therapy with erythropoietin has a positive effect on the number of germ cells. MATERIALS AND METHODS: Erythropoietin (Eprex) 100 IU/kg were administered subcutaneously weekly for 3 months prior to surgery in two cryptorchid boys, 6 months old and 1 year 9 months old, respectively, with renal function impairment. RESULTS: The number of spermatogonia per tubular cross section in testicular biopsies was unusually high in both erythropoietin- treated cryptorchid cases compared to the control material of biopsies from the undescended testes of 698 cryptorchid patients and compared to the normal values. CONCLUSION: There are several hypothetic mechanisms that can explain the elevated number of spermatogonia seen in our erythropoietin treated cryptorchid patients. Erythropoietin may have a positive effect on germ cell proliferation in cryptorchidism. PMID- 11423743 TI - In vitro fertilization. state of the art anno 2000. PMID- 11423744 TI - Testicular autotransplantation - the Monchengladbach experience. PMID- 11423745 TI - Experience with laparoscopy in nonpalpable testis. PMID- 11423746 TI - Testicular function of young men treated in childhood for cryptorchidism. PMID- 11423747 TI - Incidence of cryptorchidism in Finnish boys. PMID- 11423748 TI - Pathogenetic aspects of IgA nephropathy. PMID- 11423749 TI - The complement system in renal diseases. AB - The complement system has long been recognized as having a role in immune glomerular disease. This review provides an update on this association, some strategies for the clinical testing of complement in disease, and a brief commentary on current research directions. Evidence of complement activation in glomerulonephritis comes from characteristic patterns of a decrease in the serum concentrations of specific components, some of which are virtually diagnostic of certain nephritides. These patterns are often accompanied by the presence of complement components in the glomeruli and the detection of complement breakdown products in the circulation. In certain diseases, circulating complement activating substances can be detected. Although there are over 20 complement proteins, clinical analysis is most often directed at C3 and C4, with occasional measurement of B and C5. Recently, a variety of mechanisms for complement-induced injury has been recognized. These mechanisms go far beyond simple passive lysis of erythrocytes, the earliest functional effect of complement studied. The role of such mechanisms in renal disease is just beginning to be studied. Local synthesis of complement components in the kidney may play a role both in host defense and in the promotion of interstitial inflammation and scarring. Such mechanisms will likely be defined more precisely with the availability of animals with specific complement deficiencies. Ultimately, an understanding of the role of complement in renal disease may permit specific targeted inhibition of one or more complement functions as a form of therapy. PMID- 11423750 TI - The polymorphonuclear leukocyte--a new target for erythropoietin. AB - A previous study from our laboratory has shown that erythropoietin (EPO), beside its traditional role in erythropoiesis, acts as an alleviator of oxidative stress and inflammation in chronic hemodialysis (HD) patients, conferred in part by activated polymorphonuclear leukocytes (PMNLs). To substantiate this phenomenon, the existence of EPO receptors (EPO-Rs) on PMNL membrane was examined at the transcriptional and translational levels. mRNA for EPO-R was detected in PMNLs using specific primers directed towards the extracellular region of human EPO-R cDNA. The predicted 300-bp fragment was amplified by reverse transcriptase polymerase chain reaction. Subcloning and sequence analysis revealed 100% homology of this fragment with human EPO-R. The receptor protein was detected on the surface of intact PMNLs using (125)I-EPO. The protein was further demonstrated by flow cytometric analysis using a fluorescent monoclonal anti-EPO R. The percentage of PMNLs expressing EPO-R showed a strong correlation with the level of EPO in the serum, suggesting an upregulation of the receptor by the hormone. Taken together with our recent findings that EPO attenuates the oxidative stress and inflammation contributed by PMNLs in HD patients, the detection of functional EPO-R expression in PMNLs places these cells among the nonerythroid, EPO-responsive target populations. PMID- 11423751 TI - Differential effects of circulating IgA isolated from patients with IgA nephropathy on superoxide and fibronectin production of mesangial cells. AB - BACKGROUND: IgA nephropathy (IgAN) is characterized by predominant deposition of IgA in the glomerular mesangium. Serum IgA is often elevated in patients with IgAN, and it has been postulated that it is responsible for the mesangial lesions. However, the direct effect of circulating IgA on mesangial cells is not clear. METHODS: We investigated the effects of sera and IgA which were isolated from patients with IgAN on thymidine uptake, superoxide and fibronectin production and fibronectin mRNA expression of cultured rat mesangial cells, and we compared the findings to the effects of IgA isolated from patients with non IgA mesangial proliferative glomerulonephritis (MsPGN) and normal controls. IgA was isolated with affinity chromatography using cyanogen bromide activated Sepharose 4B coupled to sheep antihuman IgA antiserum. RESULTS: Our results demonstrated that both sera and IgA from patients with IgAN dose-dependently increased mitogenesis of mesangial cells as measured by (3)H-labeled thymidine uptake. The thymidine uptake by sera and IgA isolated from patients with IgAN was significantly higher than that of sera and IgA isolated from patients with MsPGN and normal controls. Sera and IgA from patients with IgAN significantly enhanced superoxide and fibronectin production and fibronectin mRNA expression of mesangial cells. The superoxide and fibronectin production was also significantly higher as compared with patients with MsPGN and normal controls. CONCLUSIONS: Our results indicate that circulating IgA isolated from patients with IgAN is different from that of patients with MsPGN and normal controls and may potentially induce oxidative injury and production of extracellular matrix of glomerular mesangial cells in IgAN. PMID- 11423752 TI - Effect of L-carnitine supplementation in hemodialysis patients. AB - BACKGROUND/AIM: L-Carnitine is important in beta-oxidation of fatty acids. A lack of carnitine in hemodialysis patients is caused by insufficient carnitine synthesis and especially by its loss during dialysis. The aim of our study was to test the influence of carnitine supplementation on plasma lipids, red blood cell count, and metabolism of free radicals. METHODS: Twelve regularly dialyzed patients (average age 55.5 years, average dialysis treatment period 22.5 months) were given 15 mg/kg L-carnitine intravenously three times weekly (after each hemodialysis session) for 6 months. Laboratory markers of oxidative stress, lipid metabolism, and red blood cell count were measured before the supplementation and then controlled during two 3-month intervals. Nine patients were retested 3 months after the supplementation had ended. RESULTS: All supplemented patients showed increased plasma free carnitine in comparison with the pretreatment values (113.3 +/- 11.2 vs. 62.3 +/- 16.7 micromol/l, p < 0.001). The proportion of decreased L-carnitine values after hemodialysis was reduced from 79 to 22%. Plasma total cholesterol (4.66 +/- 0.30 mmol/l after treatment vs. 5.65 +/- 1.53 mmol/l before treatment, p < 0.05) and LDL cholesterol (1.74 +/- 0.86 vs. 2.81 +/ 1.43 mmol/l, p < 0.05) decreased. The albumin concentration significantly increased from 34.8 +/- 7.3 to 46.0 +/-5.4 g/l (p < 0.05). Intraerythrocyte reduced glutathione increased from 1.65 +/- 0.25 to 2.23 +/- 0.16 mmol/l (p < 0.001), and the plasma antioxidant capacity increased from 1.65 +/- 0.09 to 2.06 +/- 0.17 mmol/l (p < 0.001). At the same time, plasma malondialdehyde decreased from 4.18 +/- 0.72 to 3.07 +/- 0.35 micromol/l (p < 0.001). The erythropoietin dose could be reduced from an average value of 5,500 to 3,500 U/week. No significant changes in the above-mentioned parameters were observed in a control group of dialyzed patients without L-carnitine supplementation. CONCLUSION: Regular carnitine supplementation of hemodialysis patients can improve their lipid metabolism, protein nutrition, red blood cell count, and antioxidant status. PMID- 11423753 TI - Nutritional status and amino acids in granulocytes and plasma in patients with chronic renal disease and varying residual renal function. AB - BACKGROUND: Protein metabolism changes with loss of renal function resulting in deterioration of nutritional status. Whether changes in macroscopic (anthropometry and subjective global nutritional assessment (SGNA)) and cellular (plasma and granulocyte free amino acid concentration) nutritional status with loss of residual renal function are related is not known. METHODS: Anthropometric variables, SGNA, and blood samples were measured after a night's fast in 43 patients (age 57 years, median (range 27-77), 32 males and 11 females) with chronic renal disease. A 24-hour urine sample was collected the day before the study for calculation of creatinine clearance and protein nitrogen appearance rate. The patients were stratified according to creatinine clearance (group I: >35 ml/min/1.73 m(2), group II 35-15 ml/min/1.73 m(2), group III <15 ml/min/1.73 m(2)). RESULTS: In males a significant lower body weight (p < 0.05) and upper mid arm muscle area (p < 0.05) was found in group III compared to group I. SGNA indicated suboptimal nutritional status in 12 patients. In group I all had normal SGNA while in group II and group III, 26 and 57% respectively were malnourished. SGNA was significantly and negatively correlated to upper mid-arm muscle area (rho = -0.37, p < 0.05) and percent body fat mass (rho = -0.46, p < 0.01) and positively correlated to percent ideal body weight (rho = 0.48, p < 0.01). Nine patients with malnutrition (M) were compared with 9 well-nourished (N) patients. They were comparable with respect to creatinine clearance (M: 13 +/- 5 ml/min/1.73 m(2); N: 16 +/- 7 ml/min/1.73 m(2)), age and sex. In malnourished patients plasma concentration of nonessential amino acids was higher (23%, p < 0.05). The intracellular concentration was generally not affected except for sulfur amino acid methionine which was increased by a factor of 2.5 (p < 0.05) and taurine decreased by a factor of 1.6 (p < 0.05). CONCLUSION: Loss of renal residual function worsened both macroscopic and cellular nutritional status. SGNA correlated to objective measures of nutritional status and is clinically usable. In malnourished chronic renal patients, increased plasma concentration of nonessential amino acids was found which might indicate increased protein degradation. PMID- 11423754 TI - Suppression of experimental membranous glomerulonephritis in rats by an anti-MHC class II antibody. AB - BACKGROUND: We previously reported that idiopathic membranous nephropathy (IMN) strongly correlated with HLA-DRB1*1501-DRB5*0101-DQAI*0102-DQB1* 0602, a specific haplotype of human major histocompatibility complex (MHC), in Japanese patients. To investigate the role of MHC in the development of rat Heymann nephritis (HN), an animal model of membranous nephropathy, a monoclonal antibody (mAb) specific to rat MHC class II antigen (RT1B) was administered, and its effectiveness in inhibiting HN was assessed. METHODS: Active HN was induced in HN-sensitive Lewis rats by administering brush border proteins of rat proximal uriniferous tubules (FX1A). Rats were divided into four groups: rats treated with 1,000 micorg anti rat MHC class II mAb, rats treated with 100 microg anti-rat MHC class II mAb, rats treated with murine myeloma IgG, and rats that did not receive either FX1A or any other mAb. We examined the differences in 24-hour urinary protein excretion and serum alloantibody titers against FX1A between groups at different time intervals, and the histologic features of kidneys at the end of the study. RESULTS: HN was induced in Lewis rats by inoculation with FX1A antigen. Administration of anti-MHC class II mAb successfully lowered urinary proteins, production of anti-FX1A alloantibodies, and the development of glomerular lesions in a dose-dependent manner. CONCLUSION: The present results demonstrated that the MHC class II molecule itself is directly involved in the pathogenesis of HN, and suggest that this therapy would be any better (or less toxic) than nonselective immunosuppressants in the treatment of IMN. PMID- 11423755 TI - Salviae radix extract prevents cisplatin-induced acute renal failure in rabbits. AB - The present study was carried out to determine if salviae radix extract (SRE) exerts a beneficial effect against cisplatin-induced renal failure in rabbits. Rabbits were pretreated with SRE orally for 7 days, followed by cisplatin injection (5 mg/kg i.p.). Cisplatin injection caused a reduction in GFR, which was accompanied by an increase in serum creatinine levels. The fractional Na+ excretion was increased by cisplatin injection. PAH uptake by renal cortical slices was inhibited by the administration of cisplatin. Such changes were prevented by SRE pretreatment. Cisplatin injection increased lipid peroxidation, which was prevented by SRE pretreatment. The protective effect of SRE was supported by morphological studies. Cisplatin injection reduced renal blood flow that was not affected by SRE pretreatment. Cisplatin treatment in vitro in renal cortical slices increased LDH release and lipid peroxidation, which were prevented by 0.05% SRE. These results indicate that lipid peroxidation plays a critical role in cisplatin-induced acute renal failure. SRE exerts a protective effect against renal cell injury induced by cisplatin, and its effect may be attributed to its antioxidant action. However, the underlying mechanism by which SRE has antioxidant action remains to be defined. PMID- 11423756 TI - Selective hypoaldosteronism due to combined defects of the conversion from inactive renin to active renin and the aldosterone biosynthesis from corticosterone. AB - A 24-year-old Japanese woman with IgA nephropathy exhibited a decreased serum aldosterone level with normal plasma renin activity after toxemia of pregnancy. Our studies revealed selective hypoaldosteronism with normal adrenoglucocorticoid functions. Levels of serum corticosterone and deoxycorticosterone were normal. Resting plasma renin activity was normal, and plasma levels of total and inactive renin were increased. Rapid ACTH administration failed to stimulate any secretion of aldosterone, whereas it adequately increased serum cortisol, deoxycorticosterone, and corticosterone concentrations. Responses of both plasma renin activity and serum aldosterone level to the furosemide-posture challenge were blunted. Angiotensin II also failed to stimulate any secretion of aldosterone despite a progressive rise in blood pressure and an appropriate increase in serum corticosterone. These results suggest that combined defects of the conversion from inactive renin to active renin and aldosterone biosynthesis are the causes of selective hypoaldosteronism in our patient. PMID- 11423757 TI - Remission achieved in chronic nephropathy by a multidrug approach targeted at urinary protein excretion. AB - Regardless of the pattern of renal involvement, increased urinary protein excretion rate is the best independent predictor of progression of chronic nephropathies and short-term reduction in proteinuria has been reported to be renoprotective in the long term. Despite such evidence, however, the therapeutic target in renoprotection is almost exclusively on blood pressure control. We report the clinical course of a patient with chronic nephropathy after the institution of a multidrug treatment titrated against urinary protein excretion to achieve renoprotection. The present findings indicate that adjusting renoprotective therapy according to the decline in protein excretion in a multidrug strategy may stabilize or even reverse renal disease progression. This approach should be formally explored in prospective studies. PMID- 11423758 TI - Membranous glomerulonephritis after haematopoietic cell transplantation for multiple myeloma. AB - Renal involvement during graft-versus-host disease following haematopoietic cell transplantation for multiple myeloma has never been described. We report a case of a recipient who developed nephrotic syndrome and membranous glomerulonephritis 22 months after the graft and 6 months after cyclosporine withdrawal. Symptoms resolved when immunosuppressive therapy was reinstituted. PMID- 11423759 TI - Treatment of severe hypothyroidism reduced serum creatinine levels in two chronic renal failure patients. AB - Short-term hypothyroidism has been associated with a reversible rise in serum creatinine levels in patients with normal renal function. A remarkable decline in serum creatinine levels associated with a treatment of severe and prolonged hypothyroidism has rarely been reported so far. We present here 2 patients with chronic renal failure in whom treatment for hypothyroidism resulted in a significant and sustained reduction of their serum creatinine levels. These cases indicate that because hypothyroidism may aggravate the serum creatinine levels, TSH should be considered in screening procedures of patients with chronic renal failure presenting with recent accelerated aggravation of renal function. Hypothyroidism per se, one of its complications or one of its associated autoimmune diseases might play a role in modifying the underlying renal problem. PMID- 11423760 TI - Hypocomplementemic type II membranoproliferative glomerulonephritis in a male patient with familial lecithin-cholesterol acyltransferase deficiency due to two different allelic mutations. AB - Patients with familial lecithin-cholesterol acyltransferase (LCAT) deficiency very often show progressive glomerulosclerosis with evolution to end-stage disease. High levels of an abnormal lipoprotein (lipoprotein X) cause glomerular capillary endothelial damage. The ultrastructural study of renal biopsy specimens shows characteristic glomerular deposits of membrane-like, cross-striated structures and vacuole structures. The gene encoding for LCAT has been mapped to chromosome 16q22.1, and several mutations of this gene cause LCAT deficiency which is inherited as an autosomal recessive trait and which is characterized by corneal opacities, normochromic normocytic anemia, and renal dysfunction. Herein we report clinical features and renal histological findings concerning a 24-year old male patient with classical familial LCAT deficiency due to two different allelic mutations: a nonsense mutation inherited from the father and a missense mutation inherited from the mother. Moreover, the patient showed glomerular histological lesions and an immunofluorescent glomerular pattern typical of hypocomplementemic membranoproliferative type II glomerulonephritis (dense deposit disease). The nature of electron-dense material that characterizes dense deposit disease is still unknown, but there are suggestions that some chemical modifications might occur in the renal basement membranes. Therefore, this clinical case might induce to consider possible relations between disorders of the lipoprotein metabolism and renal dense-deposit disease. PMID- 11423761 TI - Effect of dialysis dose on nutritional status of children on chronic hemodialysis. AB - It had been suggested that larger hemodialysis (HD) doses in children could result in better appetite, higher protein intake, better nutritional status and better growth. We investigated how different HD doses affect protein intake and nutritional status of children on chronic HD. Indices of nutritional status used were normalized protein catabolic rate (nPCR) calculated by formal 3-sample urea kinetic modeling and serum albumin level. Data of 38 HD sessions in 15 stable patients (6 males, 9 females) aged 14.5 +/- 3.28 years (mean +/- SD) were analyzed. HD sessions were divided into three groups based on delivered Kt/V: group 1 (n = 5), inadequate (Kt/V < 1.3, mean 1.05 +/- 0.14); group 2 (n = 12), adequate (Kt/V = 1.3-1.6, mean 1.50 +/- 0.07) and group 3 (n = 21), high (Kt/V >1.6, mean 1.94 +/- 0.22). Mean nPCR and Kt/V per patient during the studied week were estimated for 11 patients in whom 3 HD sessions were available within the 38 sessions analyzed. Serum albumin level was adequate in all patients (43.77 +/- 2.28 g/l). Mean overall Kt/V and nPCR were 1.68 +/- 0.36 and 1.26 +/- 0.23, respectively, r = 0.430. Average nPCR differed between groups depending on Kt/V. It was lowest in group 1 (1.01 +/- 0.12 g/kg/day) where the highest correlation between nPCR and Kt/V was found (r = 0.648). nPCR was higher and similar in groups 2 (1.27 +/- 0.23 g/kg/day) and 3 (1.31 +/- 0.22 g/kg/day), with low correlation coefficients between nPCR and Kt/V in both groups (r = 0.275 and r = 0.197, respectively). A weak positive correlation (r = 0.249) between nPCR and Kt/V was found when average weekly values per patient (n = 11) were analyzed. Results of groups 1 and 2 confirm, what is already well established in adults, that adequate dialysis needs to be achieved in order to insure good protein intake. However, our data clearly show that nPCR did not increase with a further increase in delivered HD dose, i.e. Kt/V >1.6. Our results show that the nutritional status of children on chronic HD does not seem to benefit from very high HD doses (Kt/V >1.6). PMID- 11423762 TI - Hodgkin's disease and lactic acidosis. PMID- 11423763 TI - D-dimers in hemodialysis patients. PMID- 11423764 TI - Relation between serum ubiquitin levels and KT/V in chronic hemodialysis patients. PMID- 11423765 TI - Minimal-change glomerulonephritis and chronic lymphocytic leukaemia. PMID- 11423766 TI - Oxidative stress and plasma concentrations of coenzyme Q10, alpha-tocopherol, and beta-carotene in patients with a mild to moderate decrease of kidney function. PMID- 11423767 TI - Hyperaldosteronism secondary to giant cell arteritis. PMID- 11423768 TI - Transient anuria in a patient with chronic renal failure and liver affection after a single oral dose of diclofenac. PMID- 11423770 TI - Orville T. Bailey, MD. 1909-1998. PMID- 11423771 TI - Second surgery for recurrent pilocytic astrocytoma in children. AB - Pilocytic astrocytoma (PA) is the most common childhood brain tumor. In cases where the tumor progresses or recurs following primary surgical resection, the appropriate treatment is unclear. Options include chemotherapy, radiation therapy, surgical resection or a combination thereof. To analyze the utility of further surgery, we performed a retrospective, single-institution review of pediatric patients with recurrent PAs from 1990 to 1999 who were treated with a second surgical resection. Patients were excluded if they received adjuvant chemotherapy or radiation therapy. Twenty cases were identified. Tumor locations included: cerebral hemisphere (3), cerebellum (7), optic pathway/hypothalamus (5), thalamus (1) and brainstem (4). The indication for 4 surgeries included an enlarging tumor-associated cyst. At second surgery, 10 of 20 patients had a gross total resection (GTR), 2 a near total resection (NTR), and the remaining 8 patients had a subtotal resection (STR). No patients have died. Two of 10 tumors after GTR, 0 of 2 tumors after NTR, and 7 of 8 tumors after STR had second recurrence/progression at a mean of 15 months (range 4-33 months) following second surgery. The remaining 11 patients are recurrence/progression-free at a mean of 40.7 months (range 19-119 months). Surgery for tumors or midline structures rarely resulted in a GTR (1 of 10 cases). Surgery for tumors located in the cerebral hemispheres or cerebellum resulted in GTR or NTR in all cases and can result in long periods of progression-free survival without further adjuvant treatment. PMID- 11423772 TI - Spontaneous cerebellar hemorrhage due to a juvenile pilocytic astrocytoma: case report and review of the literature. AB - A 13-year-old boy presented to the emergency room with headaches and ataxia. Imaging studies revealed a cerebellar hemorrhage within a posterior fossa tumor. The patient underwent complete resection of this lesion and made a full recovery. Microscopic examination of this lesion revealed a juvenile pilocytic astrocytoma. PMID- 11423773 TI - Pneumocephalus secondary to a neck stab wound without neurologic injury in a 13 year-old girl. AB - Pneumocephalus is usually caused by injury that damages the brain meninges and thus allows air to enter the intracranial cavity. Our intention was to establish the importance of considering a stab wound in the neck as a possible cause of traumatic pneumocephalus. The paper presents the case of a 13-year-old girl who was accidentally stabbed in the neck with a kitchen knife by her brother. She had no neurological deficit but had developed headaches. An examination showed cerebrospinal fluid leaking from the neck wound and a CT scan revealed the pneumocephalus. Following surgical treatment, the patient's clinical symptoms regressed. To our knowledge, this case is the first report of the manifestation of pneumocephalus as the result of a neck stab wound in a child; overall, there are only three reported cases of pneumocephalus caused by a neck stab wound. PMID- 11423774 TI - Recurrent pyogenic meningitis: an unusual presentation in a case of neurofibromatosis-1. AB - We report a case of recurrent pyogenic meningitis with cerebrospinal fluid rhinorrhoea in a patient with neurofibromatosis-1, which has not been reported previously. PMID- 11423775 TI - Obstructive hydrocephalus following herpes simplex virus type 1 encephalitis treated by repeated third ventriculostomy. AB - A rare case of obstructive hydrocephalus due to aqueductal obstruction following neonatal herpes simplex virus type 1 encephalitis is presented. The child was treated by third ventriculostomy and vesicles were seen on the floor of the third ventricle. The stoma closed on two occasions and required repeat third ventriculostomy, which was successful in maintaining the child shunt-free. PMID- 11423776 TI - Endoscope-assisted microsurgery for treatment of a suprasellar craniopharyngioma presenting precocious puberty. AB - Being the most common childhood tumor to involve the hypothalamus and pituitary gland, craniopharyngioma, a histologically benign tumor, is usually related to growth retardation and hypogonadism. The presentation of precocious puberty is very rare for a hypothalamic craniopharyngioma. Here, we report such a case. The female patient had presented with symptoms of menarche and breast development since she was 6 years old. Hormonal therapy with a gonadotropin-releasing hormone analogue was instituted to cease precocious puberty but was unsuccessful. Magnetic resonance imaging of the sella showed a 1-cm tumor in the hypothalamic area. Through a pterional approach, the tumor was removed en bloc with endoscope assisted microsurgery. After the operation, our patient's symptoms improved, so the hormone therapy was discontinued. This extremely unique association of craniopharyngioma and precocious puberty, the causes and mechanisms involved along with the advantages of endoscope-assisted microsurgery are discussed. PMID- 11423777 TI - Open thoracic meningocele associated with amniotic band syndrome. AB - We present a case of an infant born with an open thoracic meningocele secondary to an amniotic band. Although there have been several case reports of encephalocele, anencephaly, closed meningocele and a case of tethered cord associated with amniotic band syndrome (ABS), there has yet to be a report of an open meningocele associated with ABS. The patient remained neurologically intact after the repair of his meningocele. His postoperative course was complicated by meningitis with subsequent hydrocephalus necessitating ventriculoperitoneal shunt placement. PMID- 11423778 TI - Occipital dermal sinuses: report of nine pediatric cases and review of the literature. AB - Occipital dermal sinuses (ODS) are rare dysraphic lesions resulting from defective separation of the ectoderm and neuroderm. These lesions are often diagnosed at birth and should be treated early because of potentially life threatening complications. However, the pathogenesis of these rare lesions is still poorly understood, and there is no consensus about the optimal timing of surgery. We reviewed 9 pediatric cases of ODS operated in our department since 1982 and 79 other cases published in the literature during the last 50 years. Thirty-eight percent of these had neurological infection (meningitis or abscess), 22% had purely local infection and 40% were uncomplicated at the time of diagnosis. The incidence of infection was low during the first year of life. MRI was highly valuable to study the extent of the sinus and the content of the cyst. The outcome was regularly favorable after surgery. Surgery should however be undertaken with caution on account of the intimate relationship with venous structures, aggravated by postinfectious scarring when the patient is operated late. Considering the risks associated with surgery in neonates and the low incidence of infection during the first year, we advise performing surgery at around 6 months. PMID- 11423779 TI - Clear-cell meningioma in a 22-month-old male: a case report and literature review. AB - We report a case of spinal clear-cell meningioma occurring in a 22-month-old male who presented a right limp and then refused to walk. Spinal magnetic resonance imaging demonstrated a large, intradural tumor from T(11) to L(4), which was totally excised. The patient's postoperative recovery was uneventful and 5 months after surgery he began walking again. The latest follow-up magnetic resonance imaging of the brain and spine, obtained 42 months after diagnosis, was negative for tumor recurrence. Though clear-cell meningioma is a rare form of meningioma, it should be considered in the differential diagnosis of any space-occupying lesion of the spine arising in very young children. Complete surgical removal is necessary because it is potentially aggressive and may recur. After surgery, an accurate follow-up is warranted. PMID- 11423780 TI - Temporal lobectomy in children with intractable seizures. AB - The use of temporal lobectomy for the treatment of intractable complex partial seizures of temporal lobe origin has a long and successful history. When appropriately applied in children, excellent seizure control and neurodevelopmental progress are expected. This article reviews patient selection, evaluating the surgical procedure and outcome for temporal lobectomy, specifically in the pediatric population. PMID- 11423781 TI - Venous sinus thrombosis. PMID- 11423782 TI - Subdural fluid collections in infants. PMID- 11423783 TI - Effects of morpheme boundaries on intergestural timing: evidence from Korean. AB - This paper examines the effects of morpheme boundaries on intergestural timing, and demonstrates that low-level phonetic realization is influenced by morphological structure, i.e. compounding and affixation. It reports two experiments, one using electromagnetic midsagittal articulography (EMA) and one electropalatography (EPG), examining Korean data. The results of the EMA study show that intergestural timing is less variable for adjacent gestures across the word boundary inside a lexicalized compound than inside a nonlexicalized compound, and inside a monomorphemic word than across a morpheme boundary. The EPG study (which examined the timing in palatalization of a coronal) shows that both [ti] and [ni] have more variability in gestural timing when heteromorphemic than when tautomorphemic. Furthermore, the phonetic details of gestural overlap shed light on the asymmetry on palatalization between tautomorphemic and heteromorphemic gestural sequences (e.g. ni vs. n-i), presumably driven by paradigmatic contrast and preference of overlap. In short, what emerges from two experiments is that gestures are coordinated more stably within a single lexical item (a morpheme or a lexicalized compound) than across a boundary between lexical items. In accounting for the stability of intergestural timing within a lexical entry, several hypotheses were discussed including the Phase Window, Bonding Strength, Phonological Timing and Extended Phase Window model newly proposed here. The implication is that the morphological structure may be encoded in the phonetic realization, as is the case with other linguistic structure (e.g. prosodic structure). PMID- 11423784 TI - Perception of vowel-to-vowel transitions with different formant trajectories. AB - In this paper, the perceptual effects of vowel-to-vowel transitions determined by different temporal variations of model parameters which specify the shapes of the vocal tract area function are investigated. It is shown that, (a) the method of deformation of the vocal tract area function between two targets can be perceptually important and (b) conversely, within certain limits, the time course of parameters from one state to another, and the precise synchronization of two parameters is not important for the correct identification of a vowel series. These characteristics are necessary but not sufficient to prove the existence of a phonetic gesture percept. PMID- 11423785 TI - The acoustic qualities of Bora vowels. AB - Bora contains the vowel system /i e a o i w/, where /i/ is high, central, open, unrounded and /w/ is high, back, open, unrounded. A contrast between /i/ and /u/ is exceedingly rare in the languages of the world. An opposition between a central and a back vowel which are otherwise identical is theoretically significant since it shows that the binary feature [+/- back] is too weak to encode all phonological contrasts along the front/back dimension. However, previous analyses of languages containing such oppositions are based primarily on impressionistic transcriptions without instrumental confirmation. This study contributes to the debate by providing acoustic evidence supporting the minimal contrast between /i/ and /w/ in Bora. The results indicate that these phonemes have identical F1 values, but /i/ yields higher F2 and F3 measurements than /w/, in keeping with their reported difference in backness. PMID- 11423786 TI - Developmental trends in voice onset time: some evidence for sex differences. AB - This study reports on an investigation into the voice onset time (VOT) patterns of the plosives /p b t d/ in a group of 30 children aged 7 (n = 10), 9 (n = 10) and 11 (n = 10) years. Equal numbers of girls and boys participated in the study. Each child named a series of letter objects to elicit /p b t d/ in a syllable onset position with a fixed vowel context. VOT data were examined for age, sex and plosive differences with the following hypotheses: Firstly, that there would be sex differences in the VOT patterns of preadolescent children. Secondly, that the sex differences in VOT patterns would be linked to age and development, and that these would eventually become marked by the age of 11 years, by which time adult-like VOT values should have been achieved. Finally, that the extent of sex and age differences would be dependent upon the plosive being investigated. Results indicated patterns of decrease with age in the VOT values of /p b/ for the boys, with some evidence of increases in the VOT values of /t/ for the girls. In addition, 'voiced' and 'voiceless' cognates showed a more marked bimodal distribution in the girls' VOT patterns. This bimodal distribution was investigated by examining the degree of difference between the VOT values of voiced and voiceless cognate pairs /p b/ and /t d/, and examining the effects of age, sex and cognate pair. These results indicated that more marked sex differences in the 'voiced'/'voiceless' contrast emerged between the data of the 9- and 11-year-olds, a pattern, which was more marked for the alveolar plosives. These preliminary results confirmed all three hypotheses. The findings are presented and discussed both within a developmental and sociophonetic framework. PMID- 11423787 TI - [Minimally invasive treatment of cholelithiasis in the elderly]. AB - BACKGROUND: As cholelithiasis has an incidence which is proportional to the age of the patient, it is often detected in the elderly, who have a higher operative risk. Therefore the therapeutic approach should involve low risk and low invasive techniques. The aim of this study is to evaluate the efficacy of combined laparoscopic and endoscopic treatment in biliary tract lithiasis and gallstones. METHODS: One hundred and fifty-seven patients over 60 years old affected by cholelithiasis have been treated with minimally invasive techniques. Mean age was 68+/-5.9 years (range 60-85) and male/female ratio was 1:1.5. All the patients underwent a preoperative screening (abdominal ultrasound, liver functional tests) to evaluate the risk of choledocholithiasis associated to cholelithiasis. An ultrasound examination was performed postoperatively after 15 days and then after 6 months. RESULTS: No death due to cholecystectomy or endoscopic sphincterotomy was registered, while morbility was 5.7%. Mean hospital stay was 2+/-1.67 days which reached 5.3+/-0.7 days in case of endoscopic sphincterotomy with ERCP. CONCLUSIONS: Cholecystectomy and endoscopic sphincterotomy revealed to be low risk techniques and therefore they will to become the gold-standard treatment of cholelithiasis in the elderly. PMID- 11423788 TI - [Splenectomy for hematologic disease. Mini-invasive versus traditional technique]. AB - BACKGROUND: This study aimed to compare the safety, efficacy and clinical benefits of laparoscopic splenectomy (LS) to open splenectomy (OS) in patients with hematologic disorders. METHODS: EXPERIMENTAL DESIGN: prospective study; SETTING: II Department of Surgery, Santa Maria Nuova Hospital, Reggio Emilia and III Department of Surgery, Santo Spirito Hospital Pescara; PATIENTS: 48 consecutive adult patients underwent splenectomy; 30 patients under-went LS and 18 OS. Perioperative characteristics, outcomes, complications and costs were comparatively analyzed. RESULTS: Mean age was 35.3 years in the LS group, and 40.8 in the OS group. Mean spleen size was 11.7 cm in the LS group and 15.2 cm in the OS group. Accessory spleens were found in 5 patients in the LS group and in 4 patients in the OS group; 4 conversions to laparotomy occurred in the LS group. A total of 4 complications occurred in 3 patients of the LS; 9 complications occurred in 5 patients of OS group. Mean surgical time was 141.5 minutes for LS and 89.7 minutes for OS (p<0.005). Mean postsurgical stay was 5.8 days in the LS group and 8.5 days in the OS group (p<0.005). Response rates were similar in both groups. CONCLUSIONS: LS is comparable to OS in terms of efficacy and safety and it is associated with a shorter hospital stay. LS should become the technique of choice for treatment of intractable benign hematologic disease. PMID- 11423789 TI - [The incidence of unknown common bile duct calculosis in videolaparoscopic cholecystectomy and usefulness of routine intraoperative cholangiography]. AB - BACKGROUND: The aim of this study is to show the utility of intraoperative cholangiography IOC in detecting unknown common bile duct calculosis during VLS cholecystectomy. METHODS: The General Surgery Unit of Carlo Poma Hospital in Mantova, carried out a retrospective study considering 288 VLS executed from January 1996 to May 1999. The study excluded the following categories of patients: those with an already known choledocholitiasis, those who underwent preoperative curative ERCP, and those with a preoperative cholangio RMN. RESULTS: The IOC has detected a hepatocholedochocalculosis in 31 patients (11.4% and in 7 cases there was a suspected calculosis. Twenty of the 31 patients did not present any element that could give the suspect of calculosis. In personal experience IOC showed in 11.4% an unknown hepatic choledochal calculosis after pre-operative echography routine examinations. The percentage of success of the intraoperative cholangiography was of 90%, the meantime for this procedure 15 minute. CONCLUSIONS: The routinary execution of IOC during VLC is useful also considering its low costs. PMID- 11423790 TI - [Treatment of malignant pleural mesothelioma]. AB - BACKGROUND: In this study all patients observed between January 1993 and October 1997 with malignant pleural mesothelioma (MPM) have been analyzed in order to describe the impact of treatment modality on survival. METHODS: Medical records of 56 patients with MPM (44 male, 12 female, median age = 59 yrs) were reviewed. In 34 cases the histotype was epithelial, in 4 sarcomatoid, in 4 mixed, in 3 desmoplastic, and in 11 not specified. Four treatment modalities were identified: 1) Surgery (subtotal pleurectomy) = 20 patients; 2) Chemotherapy = 19 patients; 3) Surgery+Chemo-therapy = 8 patients; 4) Supportive care = 9 patients. RESULTS: The median survival was: 1) Surgery = 12.4 months; 2) Chemotherapy = 7.5 months; 3) Surgery+Chemotherapy = 12 months; 4) Supportive care = 11.4 months. Using univariate analysis, 8 prognostic factors were studied (age, sex, asbestos exposure, side, histotype, performance status, stage, treatment). Among these, only the stage and the performance status had shown a prognostic value on survival (p<0.05), while the treatment modality had not significantly influenced the prognosis. Using multivariate analysis only performance status showed to be significatively associated with survival (p=0.01 and odds ratio = 1.9, I.C. 1.2 3.2). CONCLUSIONS: Despite the limits of a retrospective study, personal experience confirms the ineffectiveness of current therapeutical approaches to MPM. A better understanding of MPM is required to develop new therapeutical approaches and alter the dismal prognosis of this disease. PMID- 11423791 TI - [Pleuro-pneumo-phreno-pericardiectomy with pericardial and diaphragmatic plasty]. AB - Aim of the paper is to evaluate 43 extrapleural pneumonectomy performed from 1988 to May 2000. Criteria for extrapleural pneumonectomy were pleural biopsy by thoracoscopy, potentially completely resectable unilateral disease by computed tomography and predicted postresection forced expiratory volume >1,3 L/sec. The resections regarded 33 pleural mesothelioma, 9 pleural lung-carcinosis and 1 pleural melanoma effusion. The perioperative mortality rate was 2,2% (1 death) and morbidity 21,4%. PMID- 11423792 TI - [Combined complex surgical procedures: cardiovascular and general surgery]. AB - BACKGROUND: Aim of the paper is to evaluate the surgical indication in patients with cardiac and concomitant pathology. METHODS: In this study, are examined ten combined surgical sessions performed from 1992 at 1999, in patients affected by both cardiac and neoplastic disease with surgical indication. Eight men and two women, mean age 59.4 (range 50-68 years), eight affected by thoracic or abdominal tumour, one affected by hypersplenism and one by pulmonary cystic dysplasia with recurring pneumothorax. Anatomic resection and in two patient wedge-resection were performed except one woman who was splenectomized and one man who underwent atypical resection and pulmonary decortication. All subjects required extracorporeal circulation. RESULTS: No peri-operative deaths were observed and mean survival was 34 months (range 6-72 months). Moreover, there were no hemorrhages due to coagulation values of extracorporeal circulation. This approach proved useful both from the economic point of view and hospitalization time. CONCLUSIONS: According to personal experience this can be considered a successful therapeutic choice in selected patients. These observations suggest further considerations on combined operations surgery risk and mortality morbidity appears to be reduced by a better myocardial revascularization. PMID- 11423793 TI - [Critical ischemia of the diabetic patient]. AB - BACKGROUND: Critical ischemia of the lower limbs is the final result of diabetic arteriopathy. The surgeon is more and more forced to choose between amputation d emblee and the attempt to revascularization in very old patients and in deficient general conditions. Shall we point out some principle to address our strategy? METHODS: To answer this question we reviewed retrospectively our surgical choices and the results obtained during two years, from January 1997 to December 1998 (medium follow-up 11.3 months), at the Emergency Division of the Cannizzaro Hospital of Catania. In this period 143 diabetic patients were selected by our ambulatory. Of these, nineteen had a critical lower limb ischemia and therefore were submitted to a revascularization and/or amputation. All the revascularized patients were controlled by Doppler-sonography immediately after operation and then daily, till their discharge. Controls were done at the first, third, sixth month and after one year, except for patients who complained a new objective or subjective ischemic symptomatology. RESULTS: The primary patency rate was 84.2% and the secondary patency rate was 89.4%. The amputation rate due to the procedure failure was 5%. The complications were three: two graft infections (10.5%) and one early thrombosis of a femoro-popliteal bypass graft, due to technical defect (5.2%). Perioperative mortality rate was 15.7%. CONCLUSIONS: These results, if related with those reported in the letterature about amputations, are in favour of the attempt to revascularization. PMID- 11423794 TI - [Role of the subfascial endoscopic perforator vein surgery in the treatment of venous ulcers]. AB - BACKGROUND: Limbs phlebostatic ulcers are in many cases associated with perforator reflux veins often caused by post-thromboflebitis syndrome. The origin operation consisting in perforating vein interruption, described by Linton in 1938, allowed high quality exposition of these vessels and gave successful therapeutic results only affected by complications which prolonged hospitalisation. The most unfavorable prognostic factor was the need of performing an incision in skin sites affected by dermo-hypodermic degeneration induced by phlebopathy which inhibits post-surgery cicatrization. The therapeutic importance of reflux perforator vein interruption induced vascular surgeons to improve various procedures not affected by post-surgery complications and able to excise as much incontinent vessels as possible. METHODS: By video-surgery and new surgery tools nowadays available, the Subfascial Endoscopic Perforating Veins Surgery (SEPS) was established as a feasible and valuable method. At present, there are many surgical techniques differing from each other for the tools and the subneurotic compartement from which the perforating veins start. The results obtained from 45 SEPS procedures performed on 45 patients presenting lymphodermatosclerosis or venous ulcers caused by chronic venous insufficiency have been retrospectively evaluated. RESULTS: Thirty-six patients reached ulcer healing within 4 weeks after the operation; 7 ulcers healed over a 2 month time, while for 2 patients no clinical improvement has been recorded yet. CONCLUSIONS: SEPS is a valid alternative to the Linton procedure and provides remarkable advantages both in economic and clinical terms since it yields a more rapid lesions healing of small incisions which makes it one of the most minimally invasive surgical technique. PMID- 11423795 TI - [Use of monofilament prosthesis in inguinal hernia treatment: cost-benefit ratio]. AB - BACKGROUND: The authors aimed to demonstrate the real advantages in terms of cost and patient comfort of inguinal hernia surgery using monofilament prostheses. METHODS: A retrospective survey was carried out on two groups of patients: the first group, consisting of 1032 patients who underwent inguinal hernia surgery under general anesthetic between 1985 and 1995 at the Institute of General Surgery at the University Polyclinic of Messina, included cases of both emergency and elective surgery that did not use monofilament prosthesis. The second group, consisting of 348 patients operated under local anesthesia between 1996 and 1999 at the IV Division of General Surgery at the University Polyclinic of Messina, included cases of both emergency and elective surgery using tension-free techniques and polypropylene mesh. The numbers of recidivations and complications were compared, together with the relative costs of the methods used in both groups. CONCLUSIONS: In the light of these experimental results, it is clear that the use of biocompatible alloplastic materials in monofilament considerably reduces the risks of recidivation, without no significant increase in the number of dehiscences, infections or postoperative complications. Moreover, there was a striking reduction in costs linked not only to the shorter hospitalisation of patients and the reduced use of painkillers, but also a fall in the number of future hospital admissions owing to recidivation. PMID- 11423796 TI - [Inflammatory abdominal aortic aneurysms]. AB - The authors present a review of the literature on inflammatory abdominal aortic aneurysms. These aneurysms represent from 3 to 10% of all abdominal aortic aneurysms. Progress has occurred in the technical approach to these aneurysms, and operative morbidity and mortality have been remarkably reduced. However, the pathogenesis remains poorly understood. Early reports have considered the inflammatory aneurysm as a distinct clinical and pathological entity, whereas recent evidences suggest a common etiopathogenetic mechanism for both atherosclerotic and inflammatory aneurysms. Finally, genetic and environmental factors, such as tobacco use, may predispose certain persons to the development of non-inflammatory aneurysms and others to a wide spectrum of inflammatory reactions until inflammatory aneurysms development. The most common clinical features of these aneurysms are represented by symptoms, such as abdominal or back pain, obstructive uropathy and by an elevated erythrocyte sedimentation rate. Computed tomography (CT) allows a specific diagnosis by the typical image of soft tissue surrounding the aortic wall enhancing with contrast administration. Ultrasonography is less sensitive whereas nuclear magnetic resonance (RNM) is a promising technique. Excretory urography may suggest the diagnosis by demonstration of ureter entrapment. Surgical therapy, by a technique of limited dissection represents the definitive treatment. Evolution of fibrosis after surgery is still debated because some studies have reported complete regression of inflammation and other partial regression or persistence of fibrotic process. At present, endovascular treatment of these aneurysms is occasionally reported, although preliminary results appear satisfactory. PMID- 11423797 TI - [Diagnostic delay in post-traumatic diaphragmatic ruptures]. AB - Traumatic ruptures of the diaphragm are associated with closed chest and abdominal traumas, often the outcome of road accidents, with an incidence varying from 1 to 5% depending on the series. Diagnosis is frequently late if the possibility of breakages in two stages is also considered. This delay in diagnosis is encountered in a variable percentage of cases, between 9.5 and 60%. The clinical case of a post-traumatic ruptures of the diaphragm diagnosed 26 years after a car accident is described. PMID- 11423798 TI - [Post-traumatic fracture and migration in the pulmonary artery of the catheter of a totally implantable venous access device. Unusual complication]. AB - The case of a 59 year-old patient, who sustained a post-traumatic fracture of the silastic catheter of his totally implantable venous access device that migrated in the right pulmonary artery, is reported. The venous device was placed six months earlier for the treatment of metastatic spread of a primary unknown adenocarcinoma. The venous device was placed on the left side in consideration of a recent right supraclavicular node biopsy. The catheter was introduced through the left internal jugular vein and its peripheral end was positioned subcutaneously across the clavicle to be connected to the port chamber placed in the infraclavicular region. The accidental fracture of the catheter was attributed to a closed trauma occurred during the transport of a refrigerator on the homolateral shoulder. Treatment involved extraction of the migrated fragment through a percutaneous transfemoral angioradiological procedure. A few days later the chamber was removed and a new totally implantable venous access device was placed on the other side. PMID- 11423799 TI - [Cystic tumours of the pancreas]. AB - The authors illustrate 2 cases of cystic pancreatic neoplasms recently observed at the 2nd Surgical Clinic of IRCCS Policlinico San Matteo in Pavia. Having emphasised the rarity of this pathology, they describe the specific nature of the clinical condition and symptoms which may remain unchanged for a long period. They then highlight the most significant radiological aspects, paying special attention to differential diagnosis. Surgery is indicated as the treatment of election, correlated to the topography of the lesion, the anatomic and histological nature of the disease, especially mucinous, and the patient s general conditions. PMID- 11423800 TI - Relapsing acute pancreatitis due to duodenal duplication in an 8-year-old child. Case report. AB - The authors present the diagnostic and therapeutic evolution of a case of duodenal duplication complicated by acute recurring pancreatitis in a eight-year old boy. Abdominal pain and vomiting represented important clinical signs associated with a previous history of inexplicable episodes of acute pancreatitis. Abdominal US scan and NMR were able to diagnose the duodenal duplication preoperatively. Initial treatment was conservative in the attempt to resolve the acute pancreatitis episode. Partial exeresis and derivation of the duplication in the duodenal lumen was subsequently performed. Diagnosis was confirmed by histological examination. Currently, at 6 months after intervention, the patient is clinically asymptomatic and negative at US scan follow-up. PMID- 11423801 TI - Successful transarterial embolization of idiopathic renal arteriovenous fistula. AB - A case of idiopathic renal arteriovenous fistula in a 46-year-old woman presenting intensive right renal colic associated to massive hematuria is reported. The renal arteriography confirmed the diagnosis and embolization of the fistula was performed. The transarterial embolization was successful no recurrence is observed after one year follow-up. PMID- 11423802 TI - [Post-traumatic right diaphragmatic hernia]. PMID- 11423803 TI - Use of plain radiography to screen for cervical spine injuries. AB - STUDY OBJECTIVE: Standard radiographic screening may fail to reveal any evidence of injury in some patients with spinal injury. The purposes of this investigation were to document the efficacy of standard radiographic views and to categorize the frequencies and types of injuries missed on plain radiographic screening of the cervical spine. METHODS: All patients with blunt trauma selected for radiographic cervical spine imaging at 21 participating institutions underwent a standard 3-view series (cross-table lateral, anteroposterior, and odontoid views), as well as any other imaging deemed necessary by their physicians. Injuries detected with screening radiography were then compared with final injury status for each patient, as determined by review of all radiographic studies. RESULTS: The study enrolled 34,069 patients with blunt trauma, including 818 patients (2.40% of all patients; 95% confidence interval [CI] 2.40% to 2.40%) having a total of 1,496 distinct cervical spine injuries. Plain radiographs revealed 932 injuries in 498 patients (1.46% of all patients; 95% CI 1.46% to 1.46%) but missed 564 injuries in 320 patients (0.94% of all patients; 95% CI 0.94% to 0.94%). The majority of missed injuries (436 injuries in 237 patients [representing 0.80% of all patients]; 95% CI 0.80% to 0.80%) occurred in cases in which plain radiographs were interpreted as abnormal (but not diagnostic of injury) or inadequate. However, 23 patients (0.07% of all patients; 95% CI 0.05% to 0.09%) had 35 injuries (including 3 potentially unstable injuries) that were not visualized on adequate plain film imaging. These patients represent 2.81% (95% CI 1.89% to 3.63%) of all injured patients with blunt trauma undergoing radiographic evaluation. CONCLUSION: Standard 3-view imaging provides reliable screening for most patients with blunt trauma. However, on rare occasions, such imaging may fail to detect significant unstable injuries. In addition, it is difficult to obtain adequate plain radiographic imaging in a substantial minority of patients. PMID- 11423804 TI - Use of flexion-extension radiographs of the cervical spine in blunt trauma. AB - STUDY OBJECTIVE: Flexion-extension (F/E) radiographs of the cervical spine are often used in patients with blunt trauma when the evaluating physician remains concerned about bony or ligamentous injuries despite negative or nondiagnostic standard radiographs. The use of this approach has never been addressed in a large prospective study. We sought to determine the clinical factors associated with ordering F/E views and the incidence of diagnostic F/E films in patients with a normal 3-view cervical spine series. METHODS: Patients with blunt trauma selected for radiographic cervical spine imaging at 21 participating institutions in the National Emergency X-Radiography Utilization Study project underwent standard 3-view (cross-table lateral, anteroposterior, and odontoid views) series, as well as any other imaging deemed necessary by their physicians. Injuries detected by means of screening radiography were then compared with final injury status for each patient, as determined by review of all radiographic studies. Patients who underwent F/E views were analyzed separately. RESULTS: Of 818 patients ultimately found to have cervical spine injury, 86 (10.5%) underwent F/E testing. Two patients sustained stable bony injuries detected only on F/E views. Four other patients had a subluxation detected only on F/E views, but all had other injuries apparent on routine cervical spine imaging. CONCLUSION: F/E imaging adds little to the acute evaluation of patients with blunt trauma. Other approaches, including magnetic resonance imaging, computed tomography, or delayed F/E, in the presence of specific clinical concerns would seem to provide a more reasonable approach to adjunctive imaging. PMID- 11423805 TI - Epidemiology of cervical spine injury victims. AB - STUDY OBJECTIVE: We sought to characterize demographics and injury patterns among patients undergoing emergency department cervical spine radiography for blunt traumatic injury. METHODS: All patients with blunt trauma undergoing cervical spine radiography at 21 centers were enrolled in this prospective, observational study. Patients' date of birth, age, sex, and ethnicity were noted before cervical spine radiography. RESULTS: Demographic factors associated with cervical spine injury, present in 818 of 33,922 patients, included the following: age of 65 years or older (relative risk [RR] 2.09; 95% confidence interval [CI] 1.77 to 2.59); "other" ethnicity (RR 1.79, 95% CI 1.46 to 2.19); male sex (RR 1.72, 95% CI 1.48 to 2.00); and white ethnicity (RR 1.50, 95% CI 1.31 to 1.72). Hispanic ethnicity (RR 0.64, 95% CI 0.51 to 0.79), female sex (RR 0.58, 95% CI 0.50 to 0.67), black ethnicity (RR 0.55, 95% CI 0.45 to 0.66), and age of less than 18 years (RR 0.39, 95% CI 0.27 to 0.55) were associated with reduced risk of cervical spine injury. CONCLUSION: Among patients undergoing ED cervical spine radiography, cervical spine injury is more common among the elderly, male subjects, and patients of white or "other" ethnicity. Because cervical spine injury occurs in patients in all demographic categories, however, this information cannot be used to select individual patients who should or should not undergo imaging. PMID- 11423806 TI - Distribution and patterns of blunt traumatic cervical spine injury. AB - STUDY OBJECTIVE: Previous studies of cervical spine injury involve individual institutions or special populations. There is currently little reliable information regarding natural cervical spine injury patterns after blunt trauma. This substudy of the National Emergency X-Radiography Utilization Study project was designed to accurately assess the prevalence, spectrum, and distribution of cervical spine injury after blunt trauma. METHODS: We prospectively enrolled all patients with blunt trauma undergoing cervical spine radiography at 21 diverse institutions. Injury status was determined by review of all radiographic studies obtained on each patient. For each individual injury, we recorded which specific films revealed the injury, the level and location of injury on each vertebra, and the age and sex of the patient. RESULTS: Of 34,069 enrolled patients with blunt trauma, 818 (2.4%) individuals had a total of 1,496 distinct cervical spine injuries to 1,285 different cervical spine structures. The second cervical vertebra was the most common level of injury (286 [24.0%] fractures, including 92 odontoid fractures), and 470 (39.3%) fractures occurred in the 2 lowest cervical vertebrae (C6 and C7). The vertebral body, injured in 235 patients, was the most frequent site of fracture. Nearly one third of all injuries (29.3%) were considered clinically insignificant. CONCLUSION: Cervical spine injuries occur in a small minority of patients with blunt trauma who undergo imaging. The atlantoaxial region is the most common site of injury, and the sixth and seventh vertebrae are involved in over one third of all injuries. Other spine levels are much more commonly involved than has previously been appreciated. A substantial minority of radiographically defined cervical spine injuries are of little clinical importance. PMID- 11423807 TI - Test performance of the individual NEXUS low-risk clinical screening criteria for cervical spine injury. AB - BACKGROUND: The National Emergency X-Radiography Utilization Study (NEXUS) recently validated the ability of a decision instrument to define a population with an extremely low risk of cervical spine injury (CSI) after blunt trauma. It is unclear whether each of the 5 individual criteria is necessary for the decision instrument to maintain its high sensitivity. METHODS: NEXUS was a prospective observational study at 21 emergency departments, which enrolled all patients with blunt trauma for whom cervical spine radiographs were ordered. In this substudy, we examined the NEXUS database to determine the contribution of each of the 5 individual low-risk clinical criteria to the overall sensitivity of the decision instrument. RESULTS: All but 8 of 818 patients with CSI, and all but 2 of 578 patients with significant CSI, were identified by using the decision instrument. A substantial number of patients with CSI (236/818 [29%]) and patients with significant CSI (175/578 [30%]) met only 1 of the 5 non--low-risk criteria, and each of the 5 criteria was the only indicator of non--low-risk status in at least 8 patients with CSI and at least 5 patients with significant CSI. CONCLUSION: Because each of the 5 low-risk criteria was the only marker of non--low-risk status in at least a few patients with significant CSI, modification of the overall NEXUS decision instrument by eliminating any one of the criteria would markedly reduce sensitivity and make the instrument unacceptable for clinical use. PMID- 11423808 TI - Football protective gear and cervical spine imaging. AB - STUDY OBJECTIVE: We sought to assess the effect of football protective gear on the cervical spine radiographic evaluation of adult male subjects. METHODS: The study used a prospective, randomized, matched-pairs, observational design. Subjects served as their own control subjects, with cross-table lateral and open mouth odontoid cervical spine radiographs. Radiographs were obtained with protective head and shoulder equipment (pads group) and without protective equipment (no pads group). Two emergency physicians and 2 neuroradiologists reviewed study radiographs. Physicians assessed radiographic views for adequate cervical spine visualization to the C7-T1 level and the odontoid and related structures. Comparison of radiographic readings for the pads and no pads groups used the McNemar exact test. A McNemar test of equality of paired proportions was used to estimate a population of 20 paired individuals to detect a significant outcome difference. RESULTS: Zero percent of the pads group's cross-table lateral structures were adequately visualized by all 4 reviewers (reviewer unanimity decision) compared with 25% of the no pads group's cross-table lateral films (between-group difference 25%; 95% confidence interval [CI] 6.0 to 44). When 3 of 4 reviewers noted adequate visualization (reviewer majority decision), 0% of the pads group's cross-table lateral structures were adequately visualized versus 40% of the no pads group's cross-table lateral radiographs (between-group difference 40%; 95% CI 19 to 62). With reviewer unanimity, 25% of the pads group's open mouth odontoid structures were visualized versus 45% of the no pads group's open mouth odontoid structures (between-group difference 20%; 95% CI -8.9 to 49). With reviewer majority analysis, 35% of the pads group's odontoid radiographs were adequately visualized versus 75% of the no pads group's open-mouth odontoid radiographs (between-group difference 40%; 95% CI 12 to 68). CONCLUSION: Football head and shoulder protective equipment appears to be an impediment to cervical spine radiographic visualization. Guidelines for players' cervical spine imaging should incorporate procedures for removal of equipment before initial radiographic evaluation. PMID- 11423809 TI - Defining radiographic criteria for flexion-extension studies of the cervical spine. AB - STUDY OBJECTIVE: We sought to describe and measure 3 radiographic variables in normal male volunteers and determine whether these variables could be useful in establishing more objective radiographic criteria for evaluation of flexion extension studies of the cervical spine. In addition, we hypothesized that patients with a normal cervical spine should not have greater than 2 mm of subluxation present with flexion or extension. METHODS: A prospective, observational study of normal male volunteers between the ages of 18 and 40 years was performed. We obtained radiographs of all participants in neutral, flexion, and extension positions and measured the amount of subluxation and interspinous distance, as well as the degree of vertebral angulation between C3 and C7. RESULTS: One hundred male volunteers were included in the study. Subluxation during flexion (compared with neutral) was greater than 2 mm in none of the participants at each level from C3 to C7 (95% confidence interval [CI] 0 to 3.6); subluxation in extension was greater than 2 mm in one participant at one level from C3 to C4 (95% CI 0 to 5.5) and none of the participants at each of the remaining 3 levels from C4 to C7 (95% CI 0 to 3.6). Comparing flexion with the neutral position, the mean vertebral angulation and SD were 24.2 degrees and 13.3 degrees, and the 95% certainty interval (CEI) was -1.9 to 50.2. Comparing extension with the neutral position, the mean vertebral angulation was 14.1 degrees, with an SD of 13.3 degrees, and the 95% CEI was -4.6 to 32.8. The mean change in interspinous distance between the neutral position and flexion varied from 1.2 to 4.6 mm (SD 1.7 to 2.4 mm), depending on the level of cervical spine studied. When comparing the neutral position and extension, the mean change in interspinous distance varied from 1.2 to 2.2 mm (SD 1.7 to 1.9). CONCLUSION: Currently, there are no clinically validated criteria for evaluating flexion extension studies of the cervical spine. Our study suggests that subluxation greater than 2 mm in men 18 to 40 years of age may be a useful variable for further study as an indicator of ligamentous injury. Interspinous distance and vertebral angulation appear less likely to have useful clinical application. PMID- 11423810 TI - Outcome of patients experiencing cardiac arrest with carbon monoxide poisoning treated with hyperbaric oxygen. AB - STUDY OBJECTIVE: We sought to examine the outcome of a subgroup of patients with extreme carbon monoxide (CO) poisoning, specifically those discovered in cardiac arrest, resuscitated, and subsequently treated with hyperbaric oxygen (HBO(2)). Opinions of hyperbaric medicine physicians regarding the treatment of such patients were also sought. METHODS: Records of patients treated with HBO(2) for acute CO poisoning at Virginia Mason Medical Center in Seattle from September 1987 to August 2000 were reviewed. Those who were resuscitated from cardiac arrest in the field before HBO(2) treatment were selected for detailed analysis. Patient demographic data and information regarding circumstances of the poisoning, resuscitation, HBO(2) treatment, and subsequent course were extracted and collated. In addition, a postal survey of medical directors of North American HBO(2) treatment facilities regarding opinions about the management and outcome of such patients was performed. RESULTS: A total of 18 patients were treated with HBO(2) after resuscitation from CO-associated cardiac arrest. They included 10 female and 8 male patients ranging in age from 3 to 72 years. Sources of CO included house fires (10 patients) and automobile exhaust (8 patients). Patient carboxyhemoglobin levels averaged 31.7%+/-11.0% (mean+/-SD), and arterial pH averaged 7.14+/-0.19. Presenting cardiac rhythm was a bradydysrhythmia in 10 of 18 patients. HBO(2) treatment was administered an average of 4.3 hours after poisoning (< or = 3 hours in 10 patients and < or = 6 hours in 15 patients). Despite this, all 18 patients died during their hospitalizations. Medical directors of hyperbaric treatment facilities estimated a 74% likelihood of survival for a hypothetical patient with this presentation. CONCLUSION: In this consecutive case series, cardiac arrest complicating CO poisoning was uniformly fatal, despite administration of HBO(2) therapy after initial resuscitation. Survey results suggest that physician education regarding this subset of CO poisoned patients is needed. The prognosis of this condition should be considered when making triage and treatment decisions for patients poisoned to this severity. PMID- 11423811 TI - A pilot study to assess the safety of dobutamine stress echocardiography in the emergency department evaluation of cocaine-associated chest pain. AB - STUDY OBJECTIVE: Chest pain in the setting of cocaine use poses a diagnostic dilemma. Dobutamine stress echocardiography (DSE) is a widely available and sensitive test for evaluating cardiac ischemia. Because of the theoretical concern regarding administration of dobutamine in the setting of cocaine use, we conducted a pilot study to assess the safety of DSE in emergency department patients with cocaine-associated chest pain. METHODS: A prospective case series was conducted in the intensive diagnostic and treatment unit in the ED of an urban tertiary-care teaching hospital. Patients were eligible for DSE if they had used cocaine within 24 hours preceding the onset of chest pain and had a normal ECG and tropinin I level. Patients exhibiting signs of continuing cocaine toxicity were excluded from the study. All patients were admitted to the hospital for serial testing after the DSE testing in the intensive diagnostic and treatment unit. RESULTS: Twenty-four patients were enrolled. Two patients had inadequate resting images, one DSE was terminated because of inferior hypokinesis, another DSE was terminated because of a rate-related atrial conduction deficit, and 1 patient did not reach the target heart rate. Thus, 19 patients completed a DSE and reached their target heart rates. None of the patients experienced signs of exaggerated adrenergic response, which was defined as a systolic blood pressure of greater than 200 mm Hg or the occurrence of tachydysrhythmias (excluding sinus tachycardia). Further suggesting lack of exaggerated adrenergic response, 13 (65%) of 20 patients required supplemental atropine to reach their target heart rates. CONCLUSION: No exaggerated adrenergic response was detected when dobutamine was administered to patients with cocaine related chest pain. PMID- 11423812 TI - Diagnostic use of physicians' detection of urine fluorescence in a simulated ingestion of sodium fluorescein-containing antifreeze. AB - STUDY OBJECTIVE: We sought to assess physicians' ability to accurately determine the presence or absence of sodium fluorescein (SF) in urine at a concentration corresponding to that present after ingestion of a toxic amount of commercial automotive antifreeze. METHODS: We studied 2 different urine specimen evaluation formats--one presenting isolated specimens, and the other presenting specimens grouped for comparison--to determine whether the visual clues afforded by grouped comparison aided the accuracy of the evaluation. On each study day, 3 urine specimens (1 control specimen obtained before SF administration and 2 specimens obtained after SF administration) were obtained from each of 9 or 10 volunteers. Each of these 27 or 30 urine specimens were presented sequentially and in random order to 2 emergency physicians during separate evaluation time periods. Each physician was asked to classify each specimen as fluorescent or nonfluorescent (sequential format). After a rest period, each physician, again separately, was asked to look at the same 27 or 30 urine specimens, this time all together in a test tube rack so that grouped comparisons were possible. The physicians again classified each sample as either fluorescent or nonfluorescent (grouped format). We assessed sensitivity, specificity, and accuracy of the evaluation by each presentation format (sequential or grouped). RESULTS: Mean examiner sensitivity, specificity, and accuracy for detecting the presence of SF in urine using the sequential presentation format were 35%, 75%, and 48%, respectively, whereas the same test performance indices were 42%, 66%, and 50%, respectively, when the grouped format was used. CONCLUSION: Wood's lamp determination of urine fluorescence is of limited diagnostic utility in the detection of SF ingestion in an amount equivalent to toxic ingestion of some ethylene glycol--containing automotive antifreeze products. PMID- 11423813 TI - Copperhead snakebites: clinical severity of local effects. AB - STUDY OBJECTIVE: We sought to evaluate the type and extent of local reactions after copperhead snakebites. METHODS: We performed a retrospective evaluation of copperhead snakebites in West Virginia between January 1, 1995, and September 30, 1999. A local effect scoring system was used to define a clinically significant local reaction. A bite was considered clinically significant if the bitten individual's average local effect score was 3 to 4 (range, 0 to 4). RESULTS: Ninety-two patients met the inclusion criteria; an average local effect score of 3 to 4 was documented in 33% (n=30). The foot was the most common bite location (46% of all bites); 87.5% of bites to the finger (n=8) resulted in a score of 3 to 4. Eight (36%) of 22 patients who presented to the emergency department within 2 hours of a bite and whose highest local effect score was 3 to 4 did not have their highest score until greater than 4 hours after the bite. The average length of stay (score 3 to 4) was 3.4 days compared with 1.1 days for those with a score of 3 or less. CONCLUSION: Clinically significant local effects (eg, pain requiring parenteral analgesics, ecchymosis, swelling of over one half of the bitten extremity) occurred in one third of patients in our study. The generalization of copperhead snakebites as mild or of benign clinical significance should be reconsidered. PMID- 11423814 TI - Intravenous pyridoxine-induced metabolic acidosis. AB - STUDY OBJECTIVE: Pyridoxine hydrochloride, the antidote for isonicotinic acid hydrazide (INH)--induced seizures, is available in solution at a concentration of 100 mg/mL at a pH of less than 3. Pyridoxine is often infused rapidly in large doses for INH-induced seizures. Effects of pyridoxine infusion on base deficit in amounts given for INH poisoning have not been studied in human subjects. We hypothesized that this infusion would result in transient worsening of acidosis. METHODS: We conducted a randomized, controlled crossover trial in human volunteers. Five healthy volunteers (mean age, 35 years; range, 29 to 43 years) were randomized to receive intravenous placebo (50 mL of normal saline solution) or 5 g of pyridoxine (50 mL) over 5 minutes. A peripheral intravenous catheter was established in each arm, and a heparinized venous blood sample was obtained for base deficit at baseline and 3, 6, 10, 20, and 30 minutes after infusion. After at least a 1-week washout period, the volunteers were assigned to the alternate arms of the experiments, thus acting as their own control subjects. Data were analyzed by using the 2-tailed paired t test, controlling for multiple comparisons. RESULTS: No difference was noted between groups at baseline. A statistically significant increased base deficit was noted after the pyridoxine infusion versus control at 3 to 20 minutes but not at 30 minutes (P =.1). Maximal mean increase in base deficit (2.74 mEq/L) was noted at 3 minutes. CONCLUSION: A transient increase in base deficit occurs after the infusion of 5 g of pyridoxine in normal volunteers. PMID- 11423815 TI - Protective athletic equipment and cervical spine imaging. PMID- 11423816 TI - Oral agents for the treatment of type 2 diabetes mellitus: pharmacology, toxicity, and treatment. AB - Currently available oral agents for the treatment of type 2 diabetes mellitus include a variety of compounds from 5 different pharmacologic classes with differing mechanisms of action, adverse effect profiles, and toxicities. The oral antidiabetic drugs can be classified as either hypoglycemic agents (sulfonylureas and benzoic acid derivatives) or antihyperglycemic agents (biguanides, alpha glucosidase inhibitors, and thiazolidinediones). In this review, a brief discussion of the pharmacology of these agents is followed by an examination of the adverse effects, drug-drug interactions, and toxicities. Finally, treatment of sulfonylurea-induced hypoglycemia is described, including general supportive care and the management of pediatric sulfonylurea ingestions. The adjunctive roles of glucagon, diazoxide, and octreotide for refractory hypoglycemia are also discussed. PMID- 11423817 TI - Life-threatening upper airway edema caused by a distal rattlesnake bite. AB - A 36-year-old man captured a timber rattlesnake and was accidentally envenomated in the thumb by the severed head. At a local emergency department, hypotension and confusion developed. Facial and glossal edema were also observed. Oxygen was delivered by face mask, and crystalloids and dopamine were administered. Respiratory distress developed with progressive hypoxemia. Intubation was unsuccessful because of massive glossal and epiglottic (laryngeal) edema, and an emergency cricothyrotomy was performed. High-dose antivenom therapy was administered, and mechanical ventilation was started. Recovery was rapid, and the patient was discharged from the hospital a week later. This is the first report of life-threatening upper airway edema caused by snake envenomation not in the vicinity of the head or neck. PMID- 11423818 TI - Update: Outbreak of acute febrile illness among athletes participating in Eco Challenge-Sabah 2000--Borneo, Malaysia, 2000. PMID- 11423819 TI - Three years, a thousand visits: a case study of the ultimate frequent flyer. PMID- 11423821 TI - Alternatives in the debate over the uninsured. PMID- 11423823 TI - The semantics of ketamine. PMID- 11423825 TI - Cocaine-associated acute coronary syndromes. PMID- 11423826 TI - Cocaine-associated acute coronary syndromes. PMID- 11423829 TI - Dermatoses of pregnancy. AB - The dermatoses of pregnancy can be classified into the following 3 groups: physiologic skin changes in pregnancy, dermatoses and cutaneous tumors affected by pregnancy, and specific dermatoses of pregnancy. Correct diagnosis and classification are essential for the treatment of these disorders, when necessary. Laboratory investigations are required when the diagnosis remains in question despite a careful history and thorough physical examination. A discussion with the pregnant woman about the nature of her skin condition, and the possible fetal risks associated with it, is imperative. PMID- 11423830 TI - Thimerosal in the detection of clinically relevant allergic contact reactions. AB - Thimerosal, a mercuric derivative of thiosalicylic acid, is a preservative used in several types of consumer products, including cosmetics, ophthalmic and otolaryngologic medications, and vaccines. As a result of allergic reactions and environmental concerns, its use has declined significantly during the past 2 decades. During a 5-year study at the University of Kansas Medical Center, 574 patients were patch tested to the North American Contact Dermatitis Group's standard allergen tray, which included thimerosal. The demographic data from thimerosal-allergic and nonallergic persons were compared. Statistically significant increases in thimerosal allergy were found among women, health care workers, secretaries, and cooks. Thimerosal-allergic persons were more likely to be allergic to neomycin, bacitracin, and tixocortol pivalate. Despite a high percentage of thimerosal-allergic patients in our test population, very few of these allergic reactions were found to be clinically relevant to the patient's current dermatologic condition. Using the Significance-Prevalence Index Number for thimerosal and contrasting this number with the Significance-Prevalence Index Number for other allergens on the North American Contact Dermatitis Group's standard tray, we propose that either ethyleneurea/melamine formaldehyde or bacitracin would be more useful than thimerosal as a commercially available screening allergen. PMID- 11423831 TI - Juvenile dermatomyositis: a retrospective review of a 30-year experience. AB - BACKGROUND: Children with juvenile dermatomyositis (JDMS) have variable initial presentations. OBJECTIVE: Our purpose was to evaluate the epidemiology trends, presenting clinical features, laboratory data, and outcome of patients with JDMS. METHODS: A total of 16 patients were identified at Geisinger Medical Center by a 30-year retrospective chart review. RESULTS: Sex ratio, age at diagnosis, and outcome were similar to data published in previous studies. However, certain trends were noted. The most common initial physical examination findings were an extremity rash (94%) and periungual erythema (75%). New associations of JDMS that were uncovered included the findings of pruritus (38%) and a psoriasiform scalp dermatitis (25%). Nonspecific laboratory elevations were the most common initial laboratory changes (erythrocyte sedimentation rate, lactate dehydrogenase, and aspartate aminotransferase). Tubuloreticular inclusions as found on electron microscopy of muscle biopsy specimens were present in all 3 patients tested. One patient with tubuloreticular inclusions had otherwise normal muscle biopsy findings on hematoxylin-and-eosin staining. Two of the 16 patients had cutaneous findings of JDMS but did not exhibit muscle involvement after long-term follow-up at 4 and 5 years. CONCLUSION: Our study confirms that the initial physical and laboratory findings in patients with JDMS may be nonspecific. The heliotrope rash and Gottron papules classically associated with dermatomyositis appeared less commonly than an extremity rash and periungual erythema. Creatinine kinase and aldolase levels may not be elevated on initial presentation. Pruritus, a psoriasiform scalp dermatitis, and tubuloreticular inclusions found on muscle biopsy electron microscopy should be additional factors to consider. The long term follow-up in 2 patients without muscle involvement lends support to the existence of amyopathic dermatomyositis. PMID- 11423832 TI - The association between idiopathic scoliosis and the number of acquired melanocytic nevi. AB - BACKGROUND: Several syndromes in which melanocytic nevi and scoliosis were present in the same patient have been described. No control study has been made to date to determine whether there is a relationship between these disorders. OBJECTIVE: We attempted to demonstrate the association between acquired melanocytic nevi (AMN) and idiopathic scoliosis (IS). METHODS: We studied 93 patients with IS, aged 10 to 18 years, from our hospital. Controls were randomly selected from 2 schools; Adam's forward bending test was used to exclude persons with clinical scoliosis, and the control group finally comprised 101 pupils. An observational, cross-sectional study was done. All AMN 2 mm or larger observed on the body were counted by one dermatologist. Other variables reported as risk factors in the number of nevi were also considered. Reliability of AMN counts was previously demonstrated. RESULTS: The median number of AMN was 18 (range, 10-42) in the IS group and 8 (range, 3-13) in controls (P <.001). The persons with scoliosis had more non-AMN dermatologic lesions than the controls (P <.05). Light phenotype correlated with many AMN. On multivariate analysis only scoliosis and age accounted independently for the number of AMN. CONCLUSION: IS is associated with many AMN. Multiple AMN may become a diagnostic marker for IS, and these two malformations might constitute a syndromic association. PMID- 11423833 TI - Use of the Dermatology Life Quality Index (DLQI) in a midwestern US urban clinic. AB - BACKGROUND: The Dermatology Life Quality Index (DLQI), a self-administered general dermatology quality of life instrument, was originally developed and published in a dermatology clinic at University Hospital of Wales. OBJECTIVE: Our goal was to test the feasibility of having patients answer the DLQI in a busy dermatology clinic and to find out to what extent results vary from those published in Wales. We also wanted to examine the validity of the index in terms of the correlation between DLQI scores and stage of illness (disease severity). METHODS: We administered the DLQI to 200 consecutive patients who were seen in a dermatology clinic at Indiana University Medical Center. Results were examined in light of results found by those who originated the DLQI. A pilot group of patients were given the DLQI and rated for severity of disease by means of the Dermatology Index of Disease Severity (DIDS). RESULTS: Overall, the DLQI is easy to administer and can be completed within 3 minutes. The scores in our study were compatible with those previously reported by the DLQI originators. There was a "ceiling" effect in that 11% of the patients indicated no quality of life impairment on the DLQI rating. This index shows stratification with severity of disease. CONCLUSION: The DLQI is an easy and efficient instrument for assessing quality of life in dermatology patients. Patients needed minimal assistance with the form. Our results were similar to those of the DLQI originators, and this further shows reliability and validity of the DLQI. In addition, this study further supports the use of DLQI as a quality of life instrument suitable for use in international studies. PMID- 11423834 TI - Verrucous carcinoma of the foot associated with human papillomavirus type 16. AB - BACKGROUND: We present 2 patients with verrucous carcinoma (VC) of the foot, a malignancy of unknown origin. OBJECTIVE: Molecular studies from the VCs were undertaken to determine the presence, type, and physical state of human papillomavirus (HPV) as well as the expression levels of certain oncogenes and antioncogenes. METHODS: Synthetic consensus and type-specific primers were used to determine the HPV type from both VCs via polymerase chain reaction (PCR). Verification of fragments was accomplished by means of specific isotope-labeled oligonucleotide probes. The physical state of HPV DNA was determined by two dimensional gel electrophoresis. Quantitative oncogene and antioncogene expression studies were performed with the use of reverse transcriptase PCR. RESULTS: HPV type 16 was identified in episomal and integrated forms in both tumors. Expression studies revealed increased messenger RNA levels of c-Ki-ras oncogene and the p53 antioncogene and decreased messenger RNA levels of the Rb antioncogene in both VCs. CONCLUSION: Episomal and integrated forms of HPV-16 DNA were found in VCs of the foot, along with alterations of c-Ki-ras, p53, and Rb genes. PMID- 11423835 TI - Colletotrichum species as emerging opportunistic fungal pathogens: a report of 3 cases of phaeohyphomycosis and review. AB - BACKGROUND: Numerous etiologic agents of subcutaneous phaeohyphomycosis have been reported. Colletotrichum spp, common plant pathogens, have been reported as a cause of ocular keratomycosis, but only one previous case of cutaneous disease (hyalohyphomycosis) has been attributed to this genus. OBJECTIVE: Our purpose was to describe 3 cases of subcutaneous phaeohyphomycosis due to Colletotrichum spp occurring in patients undergoing chemotherapy for hematologic malignancies. METHODS: Three cases of Colletotrichum-induced phaeohyphomycosis are reviewed. The clinical and histologic features of this infection are presented, the antifungal susceptibilities are reported, and treatment options are discussed. RESULTS: We describe the first report in which C coccodes and C gloeosporioides are implicated as etiologic agents of subcutaneous phaeohyphomycosis. Despite treatment, one patient died after the onset of visceral fungal disease. CONCLUSION: Colletotrichum spp may cause life-threatening phaeohyphomycosis in immunosuppressed patients. Prompt recognition and intervention with surgical and antifungal treatment may result in decreased morbidity and mortality associated with these infections. PMID- 11423836 TI - Release of markedly increased quantities of prostaglandin D2 from the skin in vivo in humans after the application of cinnamic aldehyde. AB - BACKGROUND: Cinnamic aldehyde is a common fragrance additive in foods and various health and beauty products. Application of cinnamic aldehyde to the skin of humans can induce cutaneous vasodilatation characterized by erythema, urticaria, and stinging. Previous studies have suggested that prostaglandins (PGs) may mediate the vasodilation, but the causative PG has not been established. We have shown that cutaneous vasodilatation induced by compounds such as sorbic acid and methylnicotinate is mediated by PGD2. OBJECTIVE: Our purpose was to determine whether cutaneous vasodilatation induced by cinnamic aldehyde is mediated by PGD2 in humans. METHOD AND RESULTS: Topical application of 1% cinnamic aldehyde to the forearms of 3 human volunteers resulted in cutaneous flushing and 25- to 42-fold increases in the levels of the major circulating metabolite of PGD2, 9alpha, 11beta-PGF2, in blood drawn from the antecubital vein draining the treated sites. There was no increase in other vasodilatory mediators, including PGE2, PGI2, or histamine. The release of PGD2 was concentration dependent. Cutaneous vasodilatation and PGD2 release were markedly decreased by the administration of aspirin, but were not significantly altered by pretreatment with the selective cyclooxygenase-2 inhibitor rofecoxib, suggesting that the formation of PGD2 is dependent on cyclooxygenase-1. CONCLUSION: The cutaneous vasodilatation induced by cinnamic aldehyde is mediated to a large extent by the release of PGD2 from a cellular source in the skin. PMID- 11423837 TI - Perianal and genital basal cell carcinoma: A clinicopathologic review of 51 cases. AB - BACKGROUND: Basal cell carcinoma (BCC) occurring on non-sun-exposed sites, especially the perianal and genital regions, is very rare. OBJECTIVE: We analyzed the incidence, clinical and pathologic features, and etiologic and prognostic factors of all non-nevoid perianal and genital BCCs diagnosed at our institution within a defined period (January 1985-September 1996). METHODS: A retrospective review was performed with the use of patient clinical records and dermatopathologic slides. Cutaneous biopsy samples were tested for the presence of human papillomavirus (HPV) by in situ hybridization using biotinylated pan-HPV and serotype-specific (6, 11, 16, 18, 31, 33, 51) probes. RESULTS: Of all non nevoid BCC syndrome cases, 51 BCCs (0.27%) were located within the perianal and genital regions. The average age of the patients was 73 years. Nine perianal BCCs occurred in men, 6 in women. Ten BCCs occurred in the pubic area, 18 on the vulva, 6 on the scrotum, and 2 on the penis. Three patients had 2 tumor sites. The average size of BCC was 1.95 cm; 29.4% were ulcerated. Seventeen patients (36%) had a history of skin cancer on sun-exposed sites and 10 (21%) had a possibly relevant associated condition. HPV was not detected in the specimens tested. Treatments included wide excision (n = 32), electrodesiccation and curettage (n = 10), Mohs micrographic surgery (n = 8), and carbon dioxide laser (n = 1). Of 30 patients with 5 years' follow-up or longer, 1 recurrence was noted 7 years after wide excision. There were no metastases. CONCLUSION: BCC of the perianal and genital skin is rare and exhibits clinical and histologic heterogeneity. Advancing age and local trauma may contribute to the pathogenesis of BCC at these sites. PMID- 11423838 TI - Physical and psychologic measures are necessary to assess overall psoriasis severity. AB - BACKGROUND: The assessment of psoriasis severity is complex and involves both the physical and psychologic assessment of the individual patient. OBJECTIVE: We compared the Salford Psoriasis Index and several other tools for assessing psoriasis severity for their abilities to assess both the physical and psychologic effects of psoriasis. METHODS: A total of 101 patients (44 women, 57 men) were assessed by means of the Salford Psoriasis Index (SPI), Psoriasis Area and Severity Index (PASI), Self-Administered PASI (SAPASI), Psoriasis Disability Index (PDI), Hospital Anxiety and Depression Scale (HADS), and Illness Perception Questionnaire (IPQ). RESULTS: The "signs" score of SPI (which measures the clinical extent of psoriasis), PASI, and SAPASI correlated well with each other (r = 0.69-0.99; P <.01). They also correlated significantly, but not as strongly, with scores of psoriasis-induced disability, the PDI and SPI "psychosocial disability" score (r = 0.46-0.51; P <.01), but not with general measures of psychologic distress. There was no significant correlation between the historical treatment, "intervention," score in SPI and either the physical or the psychologic score in the SPI. The PDI and "psychosocial disability" score of SPI correlated well with each other (r = 0.69; P <.01) as well as with the depression and anxiety subscale scores of HADS (r = 0.33 and r = 0.37; P <.01, respectively), the total number of symptoms suffered by the patient (r = 0.38; P <.01), and the belief that stress or worry were associated with psoriasis (r = 0.33; P <.01). CONCLUSION: Physical scores of psoriasis severity such as PASI, SAPASI, and the "signs" component of SPI give a partial indication of psychosocial disability caused by psoriasis. In many patients, however, the physical score does not reflect psychosocial disability. Patients should be assessed by a more holistic approach, which takes into account both physical and psychologic measurements, such as used in SPI, when assessing the severity of psoriasis. PMID- 11423839 TI - Correlation between clinical atypia and histologic dysplasia in acquired melanocytic nevi. AB - BACKGROUND: The validity of clinical and histologic criteria in identifying dysplastic nevi is controversial. Recognition of the dysplastic nevus as a distinct clinicopathologic entity requires demonstration of significant agreement between clinical atypia and histologic dysplasia. OBJECTIVE: We attempted to determine the correlation between clinical atypia and histologic dysplasia in acquired melanocytic nevi and to evaluate the sensitivity and specificity of clinical criteria for dysplastic nevi when compared with histopathologic features. METHODS: A total of 940 acquired melanocytic nevi 3 mm in diameter or larger were selected by initially choosing clinically unequivocal dysplastic and nondysplastic nevi and then, from these, histologically unequivocal dysplastic and nondysplastic lesions. The level of concordance between clinical atypia and histologic dysplasia was estimated by kappa statistics. RESULTS: Nevi were classified as clinically dysplastic (n = 499) or nondysplastic (n = 441). On the basis of histologic features, 739 were classified as dysplastic and 201 as nondysplastic. Agreement between clinical atypia and histologic dysplasia was found in 432 nevi, that is, a sensitivity of 58.4% (3-5 mm = 27.2%, >5 mm = 69.8%). Agreement between clinical and histologic criteria on the absence of dysplasia was found in 134 nevi, a specificity of 66.6% (3-5 mm = 92.4%, >5 mm = 47.9%). The kappa value was 0.17 (3-5 mm = 0.14, >5 mm = 0.10). CONCLUSION: The limited sensitivity and specificity together with the negligible kappa value indicate a poor agreement between clinical and histologic diagnoses of dysplastic nevus. The dysplastic nevus cannot be considered a distinct clinicopathologic entity because histologic dysplasia is found in a range of nevi that may or may not show clinical atypia. PMID- 11423840 TI - Phototesting in lupus erythematosus: a 15-year experience. AB - It has long been observed that sun exposure can induce or exacerbate skin lesions in patients with certain forms of lupus erythematosus. Despite the frequency of photosensitivity in these patients, the mechanism by which ultraviolet radiation alters the pathogenic course of this disease remains poorly understood. After development of standardized test methods, our group demonstrated in 1986 that skin lesions in patients with lupus erythematosus can be experimentally reproduced by UVA and UVB irradiation. In the following years, phototesting has received much attention as a valid model to study photosensitivity of different forms of lupus erythematosus and the pathogenetic mechanism of this disease. Further investigations have also made it possible to find genetic and immunologic factors associated with photosensitivity and have helped to identify the pathophysiologic steps involved in the induction of such skin lesions. We present phototesting results and clinical correlations of more than 400 patients with different forms of lupus erythematosus and discuss the recent advances in provocative phototesting. PMID- 11423841 TI - Photodynamic therapy of actinic keratoses with topical aminolevulinic acid hydrochloride and fluorescent blue light. AB - BACKGROUND: Aminolevulinic acid hydrochloride (ALA, Levulan) applied topically to actinic keratoses (AKs) leads to accumulation of the photosensitizer protoporphyrin IX, which, when activated by exposure to light, eradicates AKs. OBJECTIVE: We examined the safety and efficacy of photodynamic therapy using topical 20% ALA in a solution formulation and varying blue light doses to treat multiple AKs on the face and scalp. METHOD: This is a multicenter, investigator blinded, randomized, vehicle-controlled study. RESULTS: Thirty-six patients with clinically typical AKs were treated with 20% ALA; 14 to 18 hours later, they were irradiated with a nonlaser fluorescent blue light source. With the optimal light dose of 10 J/cm(2), 88% of the AKs completely cleared 8 weeks after a single photodynamic treatment, compared with 6% after treatment with vehicle and light. CONCLUSION: Topical ALA PDT using a nonlaser, blue light source is an effective treatment for multiple AKs. PMID- 11423842 TI - Anxiolysis with oral midazolam in pediatric patients undergoing dermatologic surgical procedures. AB - BACKGROUND: Pediatric patients undergoing dermatologic surgical procedures often experience high levels of anxiety. Oral midazolam is a short-acting benzodiazepine that can ameliorate procedure-related anxiety. OBJECTIVE: Our purpose was to determine the safety and efficacy of oral midazolam as an adjuvant anxiolytic agent for pediatric patients undergoing dermatologic surgical procedures. METHODS: A prospective series of pediatric patients undergoing dermatologic surgical procedures who received oral midazolam were monitored, and efficacy and complications were recorded. RESULTS: Oral midazolam provided good to excellent anxiolytic effects in most pediatric patients undergoing painful dermatologic surgical procedures. Because midazolam does not provide analgesic effects, local anesthesia is necessary. Complications were uncommon and minor. CONCLUSION: Oral midazolam is an effective anxiolytic agent in pediatric patients undergoing dermatologic surgical procedures. PMID- 11423843 TI - Acne scarring: a classification system and review of treatment options. AB - Acne is a common condition experienced by up to 80% of people between 11 and 30 years of age and by up to 5% of older adults. In some patients, the severe inflammatory response to Propionibacterium acnes results in permanent, disfiguring scars. Over the past several decades, numerous descriptive terms and surgical techniques have been used to diagnose the types, and improve the appearance, of scarring in those persons with acne. We propose a descriptive, simple, universally applicable acne scar classification system that includes 3 scar types: icepick, rolling, and boxcar. We also have developed an effective treatment algorithm for reconstructing and improving the appearance of acne scars including punch excision, punch elevation, subcutaneous incision (Subcision), and laser skin resurfacing. This new classification system for acne scars enables the physician to more precisely identify scar subtypes. Once the scar type has been defined, appropriate and effective treatment protocols can be developed. PMID- 11423844 TI - Position paper on family or personal leave, including pregnancy, during residency. PMID- 11423845 TI - Medical pearl: new views through the microscope. PMID- 11423846 TI - Surgical pearl: "X" marks the spot for the salivary gland biopsy. PMID- 11423847 TI - Aquagenic syringeal acrokeratoderma: report of two teenage cases. AB - Transient reactive papulotranslucent acrokeratoderma is an unusual condition of the palms that has been characterized by symmetric, hypopigmented, flat-topped papules and plaques with eccrine duct prominence that becomes more pronounced on exposure to water. Histopathologically, hyperkeratosis and dilated eccrine ostia are seen. Two teenagers with papulotranslucent acrokeratoderma are described. We propose the term aquagenic syringeal acrokeratoderma, which accurately describes the appearance after exposure to water and the prominent eccrine ducts seen clinically and histologically in the setting of acrokeratoderma. PMID- 11423848 TI - Localized syringolymphoid hyperplasia with alopecia and anhidrosis. AB - We describe a female patient with a localized form of syringolymphoid hyperplasia with alopecia and anhidrosis (SLHA). This woman is the eleventh patient with this disease so far reported in the literature. She suffered from a slowly but continuously progressing single lesion on her right ankle. It took 7 years until the diagnosis of SLHA could be established. Many divergent diagnoses were assumed and different treatments were performed during this time. The final diagnosis was established by histopathology revealing syringotropic T-cell infiltrates. Clinical features were scattered brownish papules, which formed a sharply demarcated erythematous patch lacking hairs and sweat production. The progressive course of the disease and the unresponsiveness to treatments support the current view that SLHA is a syringotropic variant of mucinosis follicularis and therefore should be viewed as a facultative precursor lesion of mycosis fungoides. In our patient, during a 7-year follow-up, no T-cell lymphoma occurred. This case emphasizes the difficulties of establishing the diagnosis of SLHA, which requires cooperation between the dermatologist and dermatopathologist. PMID- 11423849 TI - Cutis marmorata telangiectatica congenita and hypospadias: report of 4 cases. AB - Cutis marmorata telangiectatica congenita (CMTC) is an uncommon sporadic congenital vascular anomaly. Of the 111 patients with CMTC examined in our clinic during the past 25 years, 4 were found to have hypospadias. All cases were sporadic. Although CMTC has been associated with various abnormalities, to our knowledge there are no reports in the literature of concurrent hypospadias. PMID- 11423850 TI - Squamous cell carcinoma of the cervix metastatic to the skin. AB - Carcinoma of the cervix is a common neoplasm, which annually affects 50,000 women in the United States. When cervical carcinoma metastasizes, it most often involves the lung, bone, and liver; only rarely does it metastasize to the skin. We describe a patient with previously diagnosed carcinoma of the cervix who presented with a lesion on the lateral aspect of her left leg. PMID- 11423851 TI - Heterochromia of the scalp hair: a result of pigmentary mosaicism? AB - Five patients who presented stable bands of hair of a different color with respect to the surrounding hair are reported. In 4 patients this was an isolated finding. One patient also had diffuse linear skin hypopigmentation and other abnormalities. We hypothesize that these 5 cases represent a distinct type of hair heterochromia, possibly because of somatic mosaicism for genes affecting pigmentation. PMID- 11423852 TI - Linear syringocystadenoma papilliferum of the thigh. AB - Syringocystadenoma papilliferum usually arises in the head and neck region. Linear lesions occurring in other locations are rare. We report such a case occurring on the thigh, documented by multiple biopsy specimens. The case is discussed in the context of other adnexal tumors that may form linear arrangements. PMID- 11423853 TI - Treatment of resistant discoid lupus erythematosus of the palms and soles with mycophenolate mofetil. AB - Mycophenolate mofetil is an immunosuppressive drug that has recently been used to treat a variety of autoimmune and inflammatory skin diseases. Expanding the use of this agent in dermatology, we describe 2 patients with both systemic lupus erythematosus and discoid lupus erythematosus whose recalcitrant palmoplantar lesions were successfully treated with mycophenolate mofetil. PMID- 11423854 TI - PUVA therapy in lichen aureus. AB - Lichen aureus is one of the subtypes of a rare group of diseases, pigmented purpuric dermatoses. The natural course of the disease is slow evolution and slow resolution. Treatment is generally limited. We report a case of lichen aureus that responded dramatically to photochemotherapy (PUVA). PMID- 11423855 TI - Pulsed-dye laser versus conventional therapy in the treatment of warts. PMID- 11423857 TI - Milia-like idiopathic calcinosis cutis in a patient with translocation Down syndrome. PMID- 11423858 TI - Clearance in psoriasis treatment. PMID- 11423860 TI - Understanding atopic dermatitis: pathophysiology and etiology. Introduction. PMID- 11423861 TI - Atopic dermatitis versus infantile eczema. PMID- 11423862 TI - The natural history of atopic dermatitis. PMID- 11423863 TI - The genetics of atopic dermatitis: strategies, candidate genes, and genome screens. PMID- 11423864 TI - T cells in atopic dermatitis. PMID- 11423865 TI - Atopic dermatitis and the immune system: the role of superantigens and bacteria. PMID- 11423866 TI - IgE-mediated, Fc(epsilon)RI-dependent allergen presentation: a pathogenetic factor in atopic dermatitis? PMID- 11423867 TI - A role for eosinophils in atopic dermatitis. PMID- 11423868 TI - The role of keratinocytes in the pathogenesis of atopic dermatitis. PMID- 11423869 TI - Lipid abnormalities in atopic dermatitis. PMID- 11423870 TI - Disease definition and measures of disease frequency. PMID- 11423871 TI - Role of the early environment for expression of atopic dermatitis. PMID- 11423872 TI - Role of social factors in atopic dermatitis: the US perspective. PMID- 11423873 TI - Atopic dermatitis: the role of environmental and social factors, the European experience. PMID- 11423874 TI - Role of aeroallergens in atopic eczema: proof of concept with the atopy patch test. PMID- 11423875 TI - Psychologic approach to atopic skin disease. PMID- 11423876 TI - Itch in atopic dermatitis. PMID- 11423877 TI - The role of food in atopic eczema. PMID- 11423878 TI - Atopic dermatitis: predictions, expectations, and outcomes. PMID- 11423879 TI - Quality of life in atopic dermatitis. PMID- 11423880 TI - Understanding atopic dermatitis: pathophysiology and etiology. Conclusion. PMID- 11423881 TI - Urinary tract infection in childhood. A practical approach and pediatric urologists point of view. AB - Urinary tract infection is a common problem in infants and children; it is the 2nd most common infection in children; moreover, morbidity is not limited to the acute period of illness. Urinary tract infection may also result in renal scarring which has the potential for diminished renal function and hypertension. The significant morbidity from urinary tract infection in early childhood has led to our current recognition of the importance of early diagnosis, prompt antibiotic treatment, and thorough evaluation of urinary tract infections in young children with documented urinary tract infection. Especially important are the differences in urinary tract infection between adults and children that emphasize that children are not "little adults" and they need different management. The risk of renal scarring is greatest in infants and may be progressive if there is a delay in diagnosis and management of urinary tract infections in children. Normal anatomy and physiology of the urinary tract with unidirectional urinary flow and complete emptying at regular reasonable intervals should protect children from urinary tract infections. A normal child should not have urinary tract infection. The obvious goals of urinary tract infection management are to provide symptomatic relief and to prevent renal damage. In the meantime to be able to prevent the recurrences of urinary tract infection, we have to evaluate the patient more, looking for any structural or functional predisposing factors. Treatment of underlying voiding dysfunction and constipation is an essential component of successful management of urinary tract infections in children. We will elaborate on urinary tract infection in children and concentrate mainly on the pediatric urologists' point of view and our practical approach to this problem. PMID- 11423882 TI - The operative treatment of full thickness cartilage defects in the knee joint with autologous chondrocyte transplantation. AB - OBJECTIVE: The high clinical and socio-economical impact of cartilage defects and chondral degeneration is well-known. After trauma or without a known etiology, often young patients suffer from pain and a loss of function leading into a decrease of physical activity and, more severe, into long term disability and unemployment. The clinical use of autologous chondrocyte transplantation was introduced in 1994 reporting the data of a pilot study. The objective of this study is to evaluate the efficacy of this method of surgery. METHODS: Autologous chondrocyte transplantation has been established in our department since 1995 for the treatment of large, full thickness cartilage defects which can be completely covered with hyaline-like cartilage without harming the subchondral bone plate. Our first patients (n=24) all showed Grade IV lesions and an average defect size of 6.27 cm2. All but 4 of the patients had at least 1 cartilage defect related operation on the knee. RESULTS: The patients and the clinicians rating indicated an increase of a modified Cincinnati Knee score from 3.6 point pre-operation to 6.9 points after 6 months and 8.1 points at 12 months on a scale from 1 (bad) to 10 (excellent). These results support the data of an international multicenter study with almost 2000 patients. The 5 year results described by the originate authors are good to excellent in 85%-95% with an adverse event rate of 5%. CONCLUSION: Autologous chondrocyte transplantation has to be considered a safe and effective method for the treatment of large full thickness cartilage defects. Alternative treatments are symptomatical: drilling, abrasion, lavage, chondroplasty, or osteotomies. The short term results are promising but a lot of patients have to be treated for osteoarthritis as a consequence of failure with total joint arthroplasty. Osteochondral transplantations have the disadvantage of limited harvesting sites and the impairment of the subchondral bone plate. PMID- 11423883 TI - Monocyte tissue factor levels in cancer patients. AB - OBJECTIVE: The association between cancer and thromboembolic disease has been known for over a century. Increased tissue factor expression by endothelial cells, monocytes or macrophages is implicated. Thus, monocyte tissue factor measurements may reflect disease presence or progression. METHODS: Using a 2 stage kinetic chromogenic assay, monocyte tissue factor levels were assessed in normal controls (n=60), patient controls (hernia or cholecystectomy, n=60) and in patients with benign and malignant disease of the bladder (n=73), prostate (n=81), breast (n=83) and colorectum (n=62). This was performed as baseline (resting cells) and after 6 hours incubation with (stimulated) and without (unstimulated) lipopolysaccharide. Each benign disease group was sub-divided into inflammatory and non-inflammatory categories. RESULTS: The relative operating characteristic curve for the lipopolysaccharide-stimulated monocyte tissue factor assay showed sensitivity and specificity for cancer, the area under the curve being 0.71. The control groups and the benign non-inflammatory groups gave similar results and were pooled for further analysis. Each malignant group showed higher monocyte tissue factor levels than the control groups for baseline (P< 0.05) and lipopolysaccharide-stimulated cells (P< 0.05). All benign inflammatory groups apart from breast, showed increased monocyte tissue factor levels over controls for baseline (P< 0.05) and lipopolysaccharide-stimulated cells (P< 0.05). In all cases there was no significant difference between the malignant and the benign inflammatory groups. Monocyte tissue factor levels were related to tumor grade or stage, patients' survival time, serum prostate specific antigen and static bone scan images. Levels were also higher in patients with bladder cancer recurrence and in those who subsequently died. CONCLUSION: Lipopolysaccharide-stimulated monocyte tissue factor assay showed sensitivity and specificity for cancer compared to controls. Monocyte tissue factor levels are raised in malignant groups compared to controls and non-inflammatory diseases but not when compared with inflammatory conditions. Stimulated cells give better discrimination between the groups and may be useful in identifying high risk individuals. Monocyte tissue factor levels were related to tumor progression. PMID- 11423884 TI - Primary gastrointestinal cancers in the Western Region of Saudi Arabia. Is the pattern changing? AB - OBJECTIVE: To determine the age, sex and relative frequencies of various gastrointestinal malignancies in patients registered with the National Cancer Registry from the Western Region of Saudi Arabia from January 1994 till December 1997, and compare this data with previous hospital based studies about the pattern of these malignancies in Saudi Arabia. METHODS: A National Cancer Registry was established in Saudi Arabia in 1992, and since 1st January 1994 all cancer cases in Saudi Arabia have been registered with the National Cancer Registry. All National Cancer Registry data on patients with primary gastrointestinal cancers from the Western Region of Saudi Arabia from January 1994 till December 1997 was retrieved and analyzed according to ethnic origin, site, age, sex and relative frequencies of various tumors. RESULTS: Out of a total of 1833 cases with primary gastrointestinal malignancies 1207 (66%) were Saudis, while the rest were Non-Saudis. Colorectal cancer was the the most common malignancy found in both population groups accounting for 28.5% of cases in Saudis and 36% in Non-Saudis. Malignancies of liver, stomach and esophagus followed in decreasing frequencies. The mean age of the Saudi population was 58+16 years (standard deviation) with male to female ratio of 1.67:1. About 80% of the patients were above 40 years of age and the peak of onset for most of the tumors was between 50 and 70 years of age. CONCLUSIONS: This study highlights that colorectal cancer is the most common gastrointestinal malignancy seen in the Western Region of Saudi Arabia followed by hepatocellular carcinoma. This is in sharp contrast to the previous hospital based studies from Saudi Arabia and national trends as seen in the cancer incidence report from the National Cancer Registry of Saudi Arabia. The factors for this changing pattern of gastrointestinal malignancy remain to the determined. PMID- 11423885 TI - Pattern of smoking among parents of schoolboys. AB - OBJECTIVE: The objective of this study was to study the pattern, prevalence, knowledge and practice of smoking among parents of 6-15 years old schoolboys in Al-Khobar City, Saudi Arabia. METHODS: This is a cross-sectional study. The methodology included the distribution of a self-administered questionnaire, which was filled out by 2964 parents of 1482 schoolboys who satisfied the selection criteria of the study. RESULTS: The overall rate of smoking among parents of this sample was 18% (32% among fathers and 4% for mothers). A combination of heavy smoking and smoking for a long duration was the prevailing pattern of smoking. The overall sample knowledge about smoking was generally satisfactory, but smoking parents were found to have significantly higher rates of knowledge about smoking than non-smokers did. There was a significantly increased risk of smoking related diseases and symptoms among smoking parents. CONCLUSIONS: Based on the results of this study, parental smoking should be considered a major health problem in Al-Khobar City with dangerous impacts. Despite evidence of satisfactory knowledge about smoking among smokers, anti-smoking efforts should continue. However, it appears that the only sure way is to ban the import of this poison. This study may be considered as a baseline for further and larger studies in Saudi Arabia in the future. PMID- 11423886 TI - Knowledge and practice of preventive measures in small industries in Al-Khobar. AB - OBJECTIVE: The specific objective of this study was to assess the knowledge and practice of workers in small industries regarding preventive measures of occupational hazards. METHODS: Three workshops were randomly selected from each of the 8 types of industries in the industrial area. This gave a total sample of 24 workshops. From each of these, at least one worker was selected giving a total of 33 workers. All selected workers were interviewed using a standard questionnaire. A checklist on the conditions of the shop and availability of essential safety equipment was also designed. RESULTS: All workers were young non Saudis. More than half of them (58%) were smokers. Nine percent of the workers had no knowledge of preventive measures. About 12% used personal protective measures all the time while 60% did not use any. Workers were exposed to a variety of occupational hazards with injuries and accidents forming the majority (39%). CONCLUSIONS: Knowledge of workers in small industries about occupational hazards and their use of protective measures was inadequate. Workers were exposed to a variety of occupational hazards mainly accidents and injuries. PMID- 11423887 TI - Cephalometric evaluation of craniofacial variation in normal Saudi population according to Steiner analysis. AB - OBJECTIVE: The objective of this study was to describe the craniofacial pattern of Saudi ethnic groups and to compare it with accepted standards for the caucasian population according to Steiner analysis. METHODS: Lateral cephalometric radiographs of 87 Saudi students (74 male and 13 females) with balanced harmonious faces and clinically acceptable permanent occlusion, aged 21 27 years old were used. A computer-aided cephalometric analysis was performed, and the mean values and standard deviations for the various cephalometric variables were obtained. RESULTS: A comparison of the results with the Steiner standards showed that the Saudis have relatively similar skeletal relationship and dentally there was a tendency toward bimaxillary protrusion. Further, there was a decrease in the lower facial height. CONCLUSION: It was evident that even in the Saudi ethnic groups with so-called well-balanced faces, there were some fundamental variations in the craniofacial structure of the Saudi Arab when compared with Steiner norms. PMID- 11423888 TI - Assessment of reporting and recording system of communicable diseases in Jeddah Region. AB - OBJECTIVE: To determine the rate of reporting communicable diseases in Jeddah region, and to compare the recording system between the governmental and private sector. METHODS: This is a review of records study, in which the reports of communicable diseases from all hospitals and health centers (with or without cases) were studied, during the period of study; 1st to 25th international weeks 1999, and the reporting rate was calculated. A simple random sample was collected from these international weeks to evaluate the quality of recorded information. RESULTS: The reporting rate was 74%. Private hospitals have the highest rate in reporting (87%) and polyclinics have the least (67%). The recording rate was above 90% for administrative data. Personal data was complete except for patient name (76.5%), address (20%), and occupation (73%). The most prominent defect in the disease data was found to be in recording the mode of infection (13%), followed by previous vaccination (29%), date of symptoms (89%), and date of diagnosis (98%). Mode of infection was recorded in (40.5%) of cases by primary health care centers, but in polyclinics in only 1% of cases. Previous vaccination was recorded better by governmental sectors; (29%) by governmental hospital and (49%) by primary health care centers while in the private sectors the rate was (21%) by polyclinic and (25%) by private hospital. Date of symptoms was recorded in 90% of cases in all sectors except in governmental hospitals where it was only (50%). Date of diagnosis was recorded in more than (95%) in all sectors. The results show a statistical significant difference between different health sectors in recording data where government hospitals were least in recording doctor's name and in putting an official stamp in the form (p< 0.001); primary health care centers were the best in recording patient name and occupation (p< 0.001); polyclinics were least in recording mode of infection and previous vaccination (p< 0.001); for recording date of symptoms private hospitals were the best (p< 0.001). CONCLUSION: The reporting rate in Jeddah region was 74%, but its usefulness was diminished because of the incomplete, absent or incorrect personal and disease data. PMID- 11423889 TI - Hepatitis C Virus infection in Saudi Arab patients with B-cell non-Hodgkin's lymphoma. AB - OBJECTIVE: The objective of the current study is to determine the prevalence of Hepatitis C virus infection in Saudi Arab patients with B-cell non-Hodgkin's lymphoma. METHODS: Fifty-six unselected Saudi Arab patients with B-cell non Hodgkin's lymphoma were tested for the presence of Hepatitis C virus antibodies using Elisa immunoabsorbant assay 2.0. Positive and indeterminate results were subjected to confirmatory testing using RIBA-Hepatitis C virus 2.0. Two control groups were utilized for comparison; the first is a group of randomly selected general medical patients and healthy blood donors; and the 2nd is a cohort of patients with hematological neoplasms other than B-cell non-Hodgkin's lymphoma. Patients with previous history of blood transfusion or liver disease were excluded from the study. RESULTS: Twelve of the 56 B-cell non-Hodgkin's lymphoma patients (21%) tested positive for Hepatitis C virus antibodies. Only 3 out of 104 (3%) and 2 out of 41 (5%) patients tested positive for Hepatitis C virus antibodies in the first and 2nd control groups. CONCLUSION: The results of this study indicate a higher prevalence of Hepatitis C virus infection in Saudi Arab patients with B-cell non-Hodgkin's lymphoma than in the control groups. The prevalence of Hepatitis C virus infection in the 2 control groups, in turn, seems to fall within the estimated prevalence in the general population. PMID- 11423890 TI - A randomized controlled study of peritoneal closure at cesarean section. AB - OBJECTIVE: To assess the benefits or problems that may be associated with peritoneal closure at cesarean section. METHODS: A randomized-controlled study of women undergoing cesarean section in Sultan Qaboos University Hospital Maternity Unit. After the decision is taken for cesarean section, women were randomized to either repair of peritoneum using standard technique or non-repair of peritoneum. Duration of operation, maternal morbidity, blood loss assessed by post-operative hemoglobin change and requirement of transfusion, post operative infection, thromboembolic disease, and length of hospital stay were analyzed in 2 groups of patients. Sixty women were randomized into the study, 30 group A, had peritoneal closure and 30, group B, and had non-closure. RESULTS: The average duration of operation for group A was 61.9+/-12.734, and for group B was 53.56 +/-11.209 (p< 0.01 statistically significant). There was no statistically significant difference in the length of stay, estimated blood loss, the mean drop in hemoglobin, postoperative pyrexia, and wound infection rate between the 2 groups. CONCLUSIONS: Our study has confirmed the previous study findings, and has shown that there are no advantages in suturing of the peritoneum in terms of blood loss, blood transfusion, operation duration, postoperative pyrexia and wound infection. In fact non-suturing of the peritoneum was associated with shorter operation duration (p< 0.01 significant), and reduced cost. PMID- 11423891 TI - Deep vein thrombosis at King Abdul Aziz University Hospital. AB - OBJECTIVE: To study the incidence of deep vein thrombosis and pulmonary embolism at King Abdul Aziz University Hospital. To determine the risk factors, use of different diagnostic modalities, treatment given and to compare our findings with those reported in the literature. METHODS: Retrospective study which included all cases of deep vein thrombosis and pulmonary embolism admitted to the medical ward of King Abdul Aziz University Hospital during the period between January 1994 till March 1999 were analyzed. RESULTS: Total of 75 patients were diagnosed to have deep vein thrombosis with mean age of 44.16 +/- 14.5 years and male:female ratio of 1:2. Doppler ultrasound was used for the diagnosis in 56 of 75 patients (75%). Pulmonary embolism as a complication of deep vein thrombosis developed in 24 of 75 patients (32%). Prolonged immobilization was found to be the most common risk factor 17 of 75 (23%). All the patients were treated with conventional heparin followed by warfarin. CONCLUSION: As discussed, our results are comparable with those reported in the literature. Post operative patients who are anticipated to have prolonged immobilization should receive prophylactic anti coagulation with subcutaneous heparin. Thrombophillia screening should be reserved for those with recurrent deep vein thrombosis or patients with positive family history. PMID- 11423892 TI - Growth hormone deficiency in autoimmune polyglandular disease type 1. AB - This is a case report of 2 patients who were diagnosed to have autoimmune polyglandular disease type 1. Both developed mucocutaneous candidiasis, hypoparathyroidism, vitiligo, and adrenocortical insufficiency. Both were noticed to have subnormal linear growth velocity and delayed bone age. Both showed subnormal stimulated serum growth hormone values indicating growth hormone deficiency. The first case showed favorable response to growth hormone therapy. PMID- 11423893 TI - Tube enterostomy in the management of intestinal atresia. AB - A simple technique was used successfully for retraining maximum bowel length in a premature baby born with type 3A jejunal atresia. Primary end-to-end anastomosis of the tip of the dilated proximal segment to the remaining viable distal 5 cm of ileum was performed. A tube passed via the cecum proximally into the small bowel acted as a stent for the anastomosis and decompression of the bowel contents in the proximal dilated segment. This simple method may be a viable option to avoid resection of the dilated segment when the bowel length is marginal. PMID- 11423894 TI - Actinomycotic liver abscess. AB - Actinomycosis bacteria are known for their disregard to anatomical boundaries and their ability to infect organs all over the body including the liver. Here the infection is usually contained in the form of single or multiple abscesses. The clinical manifestations produced are variable even protean and only in some patients point to the right upper abdominal quadrant. However with appropriate imaging modalities and culture techniques for microaerophilic organisms accurate diagnosis is possible and specific antibiotic therapy can be initiated. We report a patient with an actinomycotic liver abscess and no apparent predisposing factor. She was diagnosed on the basis of a history of fever and right upper quadrant pain and tenderness, the abdominal ultrasound and computed tomography findings of a hypodense liver lesion and a histopathology specimen following a diagnostic and therapeutic aspiration of the liver abscess, and had an excellent response to penicillin therapy which demonstrates the nowadays possible avoidance of laparoscopic, and open surgical intervention for this condition. PMID- 11423895 TI - An unusual case of acute urinary retention in young females. AB - Urinary retention is an unusual problem in females and may be psychogenic, especially in the young. However it is important to remember that there are also organic causes for retention. We present 2 cases of an uncommon cause of urinary retention in the young female, namely hematocolpos. PMID- 11423896 TI - Massive pulmonary hemorrhage in a Saudi female with primary antiphospholipid syndrome. AB - The antiphospholipid syndrome is characterized by the presence of antiphospholipid antibodies and the association of protean clinical manifestations as a result of both venous and arterial thrombosis. Because diffuse alveolar hemorrhage leading to acute respiratory failure is a rather unusual complication of antiphospholipid antibodies, this diagnosis may be overlooked or its manifestations are attributed to another disease. Presented here is a young Saudi female with primary antiphospholipid syndrome who recovered after a stormy clinical course of acute respiratory failure in the intensive care unit. PMID- 11423897 TI - Transnasal upper gastrointestinal endoscopy in detection of gastroesophageal reflux disease induced vocal cord polyp. PMID- 11423898 TI - Effect of maternal age on fetal growth: a regional perspective. PMID- 11423899 TI - Ovarian hyperstimulation syndrome and deep vein thrombosis. PMID- 11423900 TI - Public knowledge, attitude and practice towards antibiotics use in Riyadh City. PMID- 11423901 TI - Smoking habits of students in college of Applied Medical Sciences, Saudi Arabia. PMID- 11423902 TI - Hyperhomocysteinemia: another independent vascular risk factor. PMID- 11423904 TI - Caspase cleavage of vimentin disrupts intermediate filaments and promotes apoptosis. AB - Caspases are key mediators of apoptosis. Using a novel expression cloning strategy we recently developed to identify cDNAs encoding caspase substrates, we isolated the intermediate filament protein vimentin as a caspase substrate. Vimentin is preferentially cleaved by multiple caspases at distinct sites in vitro, including Asp85 by caspases-3 and -7 and Asp259 by caspase-6, to yield multiple proteolytic fragments. Vimentin is rapidly proteolyzed by multiple caspases into similar sized fragments during apoptosis induced by many stimuli. Caspase cleavage of vimentin disrupts its cytoplasmic network of intermediate filaments and coincides temporally with nuclear fragmentation. Moreover, caspase proteolysis of vimentin at Asp85 generates a pro-apoptotic amino-terminal fragment whose ability to induce apoptosis is dependent on caspases. Taken together, our findings suggest that caspase proteolysis of vimentin promotes apoptosis by dismantling intermediate filaments and by amplifying the cell death signal via a pro-apoptotic cleavage product. PMID- 11423905 TI - Zinc inhibits p75NTR-mediated apoptosis in chick neural retina. AB - It has previously been documented that Zn2+ inhibits TrkA-mediated effects of NGF. To evaluate the ability of Zn2+ to attenuate the biological activities of NGF mediated by p75NTR, we characterized the effects of this transition metal cation on both binding and the pro-apoptotic properties of the NGF-p75NTR interaction. Binding of NGF to p75NTR displayed higher affinity in embryonic chick retinal cells than in PC12 cells. NGF induced apoptosis in dissociated cultures of chick neural retina. The addition of 100 microM Zn2+ inhibited binding and chemical cross-linking of 125I-NGF to p75NTR, and also attenuated apoptosis mediated by this ligand-receptor interaction. These studies lead to the conclusion that Zn2+ antagonizes NGF/p75NTR-mediated signaling, suggesting that the effect of this transition metal cation can be either pro- or anti-apoptotic depending on the cellular context. PMID- 11423906 TI - Combined effects of adenovirus-mediated wild-type p53 transduction, temozolomide and poly (ADP-ribose) polymerase inhibitor in mismatch repair deficient and non proliferating tumor cells. AB - Lack of p53 or mismatch repair (MR) function and scarce cell proliferation are commonly associated with tumor cell resistance to antineoplastic agents. Recently, inhibition of poly(ADP-ribose) polymerase (PARP) has been considered as a tool to overcome resistance of MR-deficient tumors to methylating agents. In the present study we demonstrated that infection with p53 expressing adenovirus (Ad-p53), enhances chemosensitivity of MR-deficient tumor cell lines to the methylating agent temozolomide (TZM), either used as single agent or, more efficiently, when combined with PARP inhibitor. Moreover, the association of Ad p53 with drug treatment induced a more pronounced growth inhibitory effect than that provoked by Ad-p53 infection only. Cells, growth arrested by p53 transduction, and then subsequently exposed to the drugs, were still highly susceptible to cytotoxicity induced by TZM and PARP inhibitor. The results suggested that this drug combination might be effective even in non-proliferating tumor cells. It is conceivable to envisage future possible strategies to enhance cytostatic or cytotoxic effects induced by Ad-p53, based on the use of TZM, alone or combined with PARP inhibitor for the therapy of resistant tumors. PMID- 11423907 TI - Interplay between the antimetastatic nm23 and the retinoblastoma-related Rb2/p130 genes in promoting neuronal differentiation of PC12 cells. AB - Increasing evidence indicates that the nm23 genes, initially documented as suppressors of metastasis progression, are involved in normal development and differentiation. We have shown previously that the murine nm23 gene enhances pheochromocytoma PC12 cells responsiveness to NGF by accelerating cell growth arrest and neurite outgrowth. The present study was aimed at elucidating the mechanisms by which nm23 controls cell proliferation and promotes neuronal differentiation. We demonstrated that nm23 modulates the expression of the Rb2/p130 gene, a negative regulator of cell cycle progression also implicated in the maintenance of the differentiated state. Furthermore, we showed that nm23-H1 mutants, defective in inhibiting the invasive phenotype, downregulate Rb2/p130 expression and inhibit NGF-induced PC12 cell differentiation. In synthesis, our results provide first evidence of interplay between the nm23 and the Rb2/p130 genes in driving PC12 cells neuronal differentiation and suggest that the antimetastatic and the differentiative nm23 functions can have similar features. PMID- 11423908 TI - Evidence of a lysosomal pathway for apoptosis induced by the synthetic retinoid CD437 in human leukemia HL-60 cells. AB - The novel synthetic retinoid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphtalene carboxylic acid (AHPN/CD437) has been proven to be a potent inducer of apoptosis in a variety of tumor cell types. However, the mechanism of its action remains to be elucidated. Recent studies suggest that the lysosomal protease cathepsin D, when released from lysosomes to the cytosol, can initiate apoptosis. In this study, we examined whether cathepsin D and free radicals are involved in the CD437-induced apoptosis. Exposure of human leukemia HL-60 cells to CD437 resulted in rapid induction of apoptosis as indicated by caspase activation, phosphatidylserine exposure, mitochondrial alterations and morphological changes. Addition of the antioxidants alpha-tocopherol acetate effectively inhibited the CD437-induced apoptosis. Measurement of the intracellular free radicals indicated a rise in oxidative stress in CD437-treated cells, which could be attenuated by alpha-tocopherol acetate. Interestingly, pretreatment of cells with the cathepsin D inhibitor pepstatin A blocked the CD437-induced free radical formation and apoptotic effects, suggesting the involvement of cathepsin D. However, Western blotting revealed no difference in cellular quantity of any forms of cathepsin D between control cells and CD437-treated cells, whereas immunofluorescence analysis of the intracellular distribution of cathepsin D showed release of the enzyme from lysosomes to the cytosol. Labeling of lysosomes with lysosomotropic probes confirmed that CD437 could induce lysosomal leakage. The CD437-induced relocation of cathepsin D could not be prevented by alpha-tocopherol acetate, suggesting that the lysosomal leakage precedes free radical formation. Furthermore, a retinoic acid nuclear receptor (RAR) antagonist failed to block these effects of CD437, suggesting that the action of CD437 is RAR-independent. Taken together, these data suggest a novel lysosomal pathway for CD437-induced apoptosis, in which lysosomes are the primary target and cathepsin D and free radicals act as death mediators. PMID- 11423909 TI - Tissue expression and subcellular localization of the pro-survival molecule Bcl w. AB - Anti-apoptotic members of the Bcl-2 family, such as Bcl-w, maintain cell viability by preventing the activation of the cell death effectors, the caspases. Gene targeting experiments in mice have demonstrated that Bcl-w is required for spermatogenesis and for survival of damaged epithelial cells in the gut. Bcl-w is, however, dispensable for physiological cell death in other tissues. Here we report on the analysis of Bcl-w protein expression using a panel of novel monoclonal antibodies. Bcl-w is found in a diverse range of tissues including colon, brain and testes. A survey of transformed cell lines and purified hematopoietic cells demonstrated that Bcl-w is expressed in cells of myeloid, lymphoid and epithelial origin. Subcellular fractionation and confocal laser scanning microscopy demonstrated that Bcl-w protein is associated with intracellular membranes. The implications of these results are discussed in the context of the phenotype of Bcl-w-null mice and recent data that suggest that Bcl w may play a role in colon carcinogenesis. PMID- 11423910 TI - Caspases mediate nucleoporin cleavage, but not early redistribution of nuclear transport factors and modulation of nuclear permeability in apoptosis. AB - In eukaryotic cells, both soluble transport factors and components of the nuclear pore complex mediate protein and RNA trafficking between the nucleus and the cytoplasm. Here, we investigated whether caspases, the major execution system in apoptosis, target the nuclear pore or components of the nuclear transport machinery. Four nucleoporins, Nup153, RanBP2, Nup214 and Tpr are cleaved by caspases during apoptosis. In contrast, the nuclear transport factors, Ran, importin alpha and importin beta are not proteolytically processed, but redistribute across the nuclear envelope independently and prior to caspase activation. Also, mRNA accumulates into the nucleus before caspases become active. Microinjection experiments further revealed that early in apoptosis, the nucleus becomes permeable to dextran molecules of 70 kD molecular weight. Redistribution of import factors and mRNA, as well as nuclear permeabilisation, occur prior to caspase-mediated nucleoporin cleavage. Our findings suggest that the apoptotic programme includes modifications in the machinery responsible for nucleocytoplasmic transport, which are independent from caspase-mediated degradation of nuclear proteins. PMID- 11423911 TI - Sensitivity of Ewing's sarcoma to TRAIL-induced apoptosis. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is able to kill transformed cells. We have studied the expression and functionality of the TRAIL apoptotic pathway in Ewing's sarcoma. We demonstrate that tumors from patients with Ewing's sarcoma express receptors TRAIL-R1 and -R2. Using a panel of nine Ewing's sarcoma cell lines TRAIL could induce apoptosis in seven cell lines. Preincubation with interferon-gamma rendered the two resistant cell lines sensitive. TRAIL was the most potent inducer of apoptosis when compared to Fas ligand or TNF. TRAIL-mediated apoptosis could be inhibited by various caspase inhibitors. No difference in the surface expression of TRAIL-receptors was observed between sensitive and resistant cell lines. Also, all cell lines had similar levels of expression of Flice-like inhibitory protein (FLIP) on immunoblot. However, the two resistant cell lines had only very low level expression of caspase 8 on RNA and protein level. In summary, we show that Ewing's sarcoma expresses receptors for TRAIL, and that cells are exquisitely sensitive to TRAIL-mediated apoptosis. These results may warrant clinical trials with TRAIL in Ewing's sarcoma once the safety of TRAIL for humans has been established. PMID- 11423912 TI - Even after UVA-exposure will nitric oxide protect cells from reactive oxygen intermediate-mediated apoptosis and necrosis. AB - Reactive oxygen species (ROS) play a pivotal role in UVA-induced cell damage. As expression of the inducible nitric oxide synthase (iNOS) is a normal response of human skin to UV radiation we examined the role of nitric oxide (NO) as a protective agent during or even after UVA1- or ROS-exposure against apoptosis or necrosis of rat endothelial cells. When added during or up to 2 h subsequent to UVA1 or ROS exposure the NO-donor S-nitroso-cysteine (SNOC) at concentrations from 100-1000 microM significantly protects from both apoptosis as well as necrosis. The NO-mediated protection strongly correlates with complete inhibition of lipid peroxidation (sixfold increase of malonedialdehyde formation in untreated versus 1.2-fold with 1 mM SNOC). NO-mediated protection of membrane function was also shown by the inhibition of cytochrome c leakage in UVA1 treated cells, a process not accompanied by alterations in Bax and Bcl-2 protein levels. Thus, the experiments presented demonstrate that NO exposure during or even after a ROS-mediated toxic insult fully protects from apoptosis or necrosis by maintaining membrane integrity and function. PMID- 11423913 TI - Inhibition of phosphatidylinositol-3 kinase/Akt or mitogen-activated protein kinase signaling sensitizes endothelial cells to TNF-alpha cytotoxicity. AB - Bovine carotid artery endothelial (BAE) cells are resistant to tumor necrosis factor-alpha (TNF), like most other cells. We examined if mitogen-activated protein (MAP) kinase and phosphatidylinositol-3 (PI3) kinase/Akt pathways are involved in this effect. In BAE cells, TNF activates MAP kinase in a MAP kinase kinase 1 (MEK1) manner and Akt in PI3-kinase-dependent manner. Pretreatment with either the MEK1 inhibitor U0126 or PI3-kinase inhibitor LY294002 sensitized BAE cells to TNF-induced apoptosis. Neither U0126 nor LY294002 pretreatment affected TNF-induced activation of NF-kappaB, suggesting that the MAP kinase or PI3 kinase/Akt-mediated anti-apoptotic effect induced by TNF was not relevant to NF kappaB activation. Both MAP kinase and PI3-kinase/Akt -mediated signaling could prevent cytochrome c release and mitochondrial transmembrane potential (Deltapsi) decrease. PI3-kinase/Akt signaling attenuated caspase-8 activity, whereas MAP kinase signaling impaired caspase-9 activity. These results suggest that TNF induced MAP kinase and PI3-kinase/Akt signaling play important roles in protecting BAE cells from TNF cytotoxicity. PMID- 11423914 TI - Apoptosis--regulation and clinical implications. PMID- 11423916 TI - Validity of DISHES 98, a computerised dietary history interview: energy and macronutrient intake. AB - OBJECTIVE: To estimate the relative validity of a computerised dietary history instrument (DISHES 98). SETTINGS: Munich and Berlin. SUBJECTS: A total of 148 persons aged 19-59 y recruited from two research centres. DESIGN: A relative validation study. Energy and macronutrient intakes obtained with DISHES 98 were compared to those assessed with 3-day weighed dietary records and with a 24 h dietary recall. RESULTS: Intakes of energy, total, saturated and monounsaturated fat, polysaccharides and alcohol were significantly higher and intake of dietary fibre was significantly lower with the 3-day records than with DISHES 98. For intakes of total, animal and vegetable protein, total carbohydrates, mono- and disaccharides and cholesterol the mean difference between DISHES 98 and the 3-day dietary records was less than 5% of the intake with DISHES 98. Pearson's correlation coefficients between DISHES 98 and 3-day records varied from 0.34 for intake of polyunsaturated fat to 0.69 for intake of disaccharides and from 0.27 for polyunsaturated fat to 0.65 for total carbohydrates between DISHES 98 and the 24 h recall. The proportion of participants classified into the same or adjacent quintile of intake varied between 66.9% for polyunsaturated fat and 90.4% for alcohol comparing DISHES 98 and 3-day records and between 60.2% for polyunsaturated fat and 78.4% for total carbohydrates comparing DISHES 98 and 24 h recalls. CONCLUSION: The observed differences between DISHES 98 and the other methods are in an acceptable range for assessing dietary intake in epidemiologic studies. PMID- 11423917 TI - Seeing mum drinking a 'light' product: is social learning a stronger determinant of taste preference acquisition than caloric conditioning? AB - OBJECTIVE: It was examined whether caloric conditioning or social learning strategies dominate in taste preference acquisition in children. The caloric learning paradigm predicts that eating or drinking artificially sweetened products, which deliver virtually no energy, will not lead to a taste preference whereas the social learning paradigm predicts that seeing important others modelling the eating and drinking of these 'light' products will induce a preference for the taste of light products in the child. DESIGN: In a 2 x 2 between subjects factorial design, the amount of energy and social modelling was varied. SETTING: The study was undertaken at primary schools in Maastricht, The Netherlands. SUBJECTS: Forty-five children participated and six children dropped out. The 39 children who completed the study (14 boys and 25 girls) had a mean age of 67 months (range 51--81, s.d. 5.6). INTERVENTIONS: Each subject took part in nine conditioning trials with an individually selected tasting yoghurt which was not preferred very much at the pre-test. RESULTS: The children in the combined caloric and social condition showed an increase in their preference for the conditioned taste which was larger than a regression-to-the-mean effect (P=0.007), whereas children in the other groups did not. CONCLUSION: Caloric and social learning combined, ie modelling the consumption of energy-rich foods or drinks, is the best way to establish taste preferences. Children more easily learn a preference for energy-rich food that is eaten by significant others than for food that is low in energy and eaten by significant others. PMID- 11423918 TI - Underestimation of percentage fat mass measured by bioelectrical impedance analysis compared to dual energy X-ray absorptiometry method in obese children. AB - OBJECTIVE: The aim of the study was to investigate whether there is a difference between body fat mass percentage measured by BIA and DXA method. DESIGN: Transversal study, randomized. SETTING: Lipid and Obesity Outpatient Clinic, Department of Pediatrics, University of Vienna, Austria. SUBJECTS: Twenty-seven children and adolescents from the Lipid and Obesity Outpatient Clinic, Department of Pediatrics, University of Vienna, were included in the study (14 boys and 13 girls between 6 and 18 y; mean age 12.6 and 13.1 y). METHODS: The body fat percentage was measured with BIA (bioelectrical impedance analyzer BIA 2000-M) and DXA (dual energy X-ray absorptiometry) methods on the same day. RESULTS: The mean difference between the body fat mass percentage measured by BIA and DXA was 4.48 with a standard deviation of 2.93. The results measured by BIA were almost always lower than that by DXA by about 12%. The lower and upper limit of the difference in 95% confidence interval was -5.64 and -3.32. By paired t-test, these results were significantly different (P<0.001). The correlation between the two measurements was 0.826. The mean percentage of body fat mass measured by BIA was 34.86+/-7.08% and by DXA 39.75+/-5.63%. The differences were not changed by age and body fat percentage but they were by sex. CONCLUSIONS: The results of the study show that the body fat percentages measured by BIA and DXA method were significantly different. This is very important because BIA technique is a routine technique for clinical purposes. Adjustments to the formula used for calculating the total fat mass in obese children and adolescence are necessary. Underestimation of body fat percentage measured by bioelectrical impedance analysis compared to dual X-ray absorptiometry method in obese children is three times higher with boys than with girls. PMID- 11423919 TI - Influence of sociodemographic factors in the prevalence of obesity in Spain. The SEEDO'97 Study. AB - OBJECTIVE: To analyse the influence of social and cultural factors in the prevalence of obesity in the Spanish adult population aged 25--60 y based on available population data. DESIGN: Pooled analysis of four cross-sectional nutrition surveys. SUBJECTS: A total of 5388 free-living subjects aged 25--60 y, respondents of the Nutritional Surveys carried out in four Spanish regions (Catalunya, Basque Country, Madrid and Valencia) from 1990 to 1994. The samples were pooled together and weighted to build a national random sample. MEASUREMENTS: Weight and height were measured on each individual by trained observers. Age, gender, educational level, occupation, habitat (rural/urban) and region were considered. Obesity was defined as body mass index > or = 30 kg/m(2). The protocol used in each survey was in accordance with the recommendations of the Spanish Society for the Study of Obesity (SEEDO). Logistic regression models were designed to analyse the influence of sociodemographic factors in the prevalence of obesity in men and women. RESULTS: The prevalence of obesity was higher in older age groups in men and women, odds ratio (OR) for every 10 y OR=1.40 (95% CI 1.39--1.41) for men and OR=1.86 (95% CI 1.85--1.87) for women. Logistic regression analysis adjusted for age showed higher obesity rates among low educated people, OR=1.80 (95% CI 1.78--1.81) in men and OR=2.36 (95% CI 2.29- 2.42) in women (P<0.001). Among men the odds ratio for the prevalence of obesity in rural areas was OR=1.87 (95% CI 1.86--1.89), compared to cities. The geographical distribution showed higher obesity rates in the southeast. CONCLUSION: This study supports that obesity is a multifactorial problem. Older women with low educational level and low income seem to be the most susceptible group to weight gain. Therefore, Public Health Programs should consider this type of environmental factor when planning strategies aimed at preventing or reducing the problem of obesity in western societies. PMID- 11423920 TI - Effect of a high monounsaturated fatty acid diet on blood pressure and glucose metabolism in women with gestational diabetes mellitus. AB - OBJECTIVE: The effect of a diet rich in monounsaturated fatty acids (MUFA) on blood pressure, glycemic control, lipids and insulin sensitivity was evaluated in women with gestational diabetes mellitus. DESIGN AND METHODS: A randomized, unpaired diet intervention was performed in 27 women with gestational diabetes mellitus in an outpatient clinic. After randomization the women received either a high-carbohydrate diet (H-CHO) or a high-MUFA diet (H-MUFA) from the 33rd gestational week of pregnancy. Outcome measures were 24 h ambulatory blood pressure, blood lipids, glycemic control and insulin sensitivity estimated by an intravenous glucose tolerance test. RESULTS: The 24 h diastolic blood pressure increased more in the H-CHO group than in the H-MUFA group (P<0.04). CONCLUSIONS: After 5 weeks of treatment with a MUFA-enriched diet, no increase in 24 h diastolic blood pressure and no adverse effects on blood lipids were seen. The favorable effect on the blood pressure by the MUFA diet is a possible non medication treatment. The H-MUFA diet had no advantage to the H-CHO diet in ameliorating the decline of insulin sensitivity in third term of pregnancy in GDM. PMID- 11423921 TI - Complementary effects of Mediterranean diet and moderate red wine intake on haemostatic cardiovascular risk factors. AB - OBJECTIVES: To compare the effect of alcohol-free Mediterranean-type diet (MD) and high-fat diet (HFD) on plasma concentration of emergent haemostatic cardiovascular risk factors (HCVRF). Also, to test if red wine supplementation modifies HCVRF, independent of diet. DESIGN, SUBJECTS AND INTERVENTION: Controlled prospective intervention study. Two groups, each of 21 healthy male university students (22+/-3.4 y), received either MD or HFD for 90 days. Between days 30 and 60, both diets were supplemented with 240 ml/day of red wine. Baseline and T30, T60 and T90-day samples were drawn. No drop out from the study was observed. SETTING: University campus and outpatient nutrition clinic. RESULTS: Volunteers on HFD at T30 had increases in pro-coagulants fibrinogen (22%), factor VIIc (9%), and factor VIIIc (4%), and decreases in natural anticoagulants antithrombin III (3%), protein C (11%) and protein S (6%) and of 20% in plasminogen activator inhibitor-1. At the same time, individuals on MD had increases in fibrinogen (4%), antithrombin III (5%), protein C (3%), protein S (2.7%), and decreases in factor VIIIc (9%), and plasminogen activator inhibitor-1 (21%). After adjusting by baseline values, MD was associated with lower plasma fibrinogen (P=0.03), factor VIIc (P=0.034) and factor VIIIc (P=0.0057) and with higher levels of protein S (P=0.013). Red wine supplementation, in both diets, resulted in decreased plasma fibrinogen (P=0.001) and factor VIIc (P=0.05), and increased tissue plasminogen activator antigen (P=0.01) and plasminogen activator inhibitor-1 antigen (P=0.0003). Wine consumption was also associated with significantly (P=0.01) divergent effects on antithrombin III: it decreased by 10% in individuals on HFD but increased slightly in those on MD. No effects of diet or wine were detected in plasma protein C and C-reactive protein. CONCLUSION: MD and moderate consumption of red wine have complementary, mostly beneficial effects on HCVRF. PMID- 11423922 TI - Fat intake is more strongly associated with lifestyle factors than with socio economic characteristics, regardless of energy adjustment approach. AB - OBJECTIVE: To compare how three different energy adjustment approaches influence the ranking of individuals on fat intake, and to examine the relation between fat intake and socio-economic, demographic and lifestyle characteristics for each energy adjustment approach. DESIGN: A cross-sectional analysis project, using a sub-sample (7055 women and 3240 men) from the Malmo Diet and Cancer Study. Dietary habits were assessed with an interview based diet history method. Fat intake was, depending on energy-adjustment method, defined as absolute intake (FATg), percentage energy from fat (FAT%), and residuals from total fat regressed on total energy (FATres). Cross-classification compared categorisation into fat intake quintiles. Logistic regression estimated, separately for each of the three approaches, the associations between high fat intake and socio-economic, demographic and lifestyle characteristics. RESULTS: Agreement in individuals' ranking was high between FAT% and FATres, but FATg differed substantially from the others. Current smoking, low level of leisure time physical activity and low alcohol intakes were, in multivariate analysis, consistently associated with risk of high fat consumption regardless of energy adjustment method. However, the associations with socio-economic characteristics varied with energy adjustment method and gender groups. CONCLUSIONS: The similarities between FAT% and FATres, in the ranking of individuals and in the association with lifestyle factors and socio-economic characteristics implies that it is possible to translate results obtained with FATres to recommendations using FAT%. The consistent lifestyle pattern across fat intake definitions (in energy adjusted models) may indicate that fat consumption is more strongly related to lifestyle factors than to socio economic characteristics. PMID- 11423923 TI - Relation between body composition and age in healthy Japanese subjects. AB - OBJECTIVE: To describe the relation between body composition and age measured by dual-energy X-ray absorptiometry (DXA) in healthy Japanese adults. DESIGN: Cross sectional study. SUBJECTS AND MEASUREMENTS: The subjects were 2411 healthy Japanese adults (males 625, females 1786, age 20--79 y) who attended the Fukuoka Health Promotion Center, Fukuoka, Japan for health check-up. Body composition was determined by DXA (QDR-2000, Hologic) for the whole body and three anatomical regions of arms, legs and trunk. RESULTS: The mean values of body mass index (BMI) and percentage fat mass (%FM) were 23.2+/-3.1 (s.d.) kg/m(2) and 21.8+/ 6.8% for males and 22.1+/-3.3 kg/m(2) and 32.0+/-7.5% for females, respectively. For males, curvilinear relations with the peaks in their forties or fifties were seen for the variables associated adiposity, ie BMI, waist and hip circumference, waist-hip ratio, total or regional fat mass (FM), %FM and ratio of trunk FM to leg FM. For females, most of these variables increased linearly in older subjects. Lean mass (LM), bone mineral content (BMC) and bone mineral density (BMD) of the whole body and appendicular LM were relatively constant until the forties and then decreased in both sexes. The rates of decrease in the total or appendicular LM were larger for males than for females, whereas those in BMC or BMD were larger for females than for males. CONCLUSIONS: This study presents the first detailed data on body composition in Japanese, which may be useful when comparing with populations of different racial and ethnic backgrounds and studying ill subjects. PMID- 11423924 TI - Does fat intake predict adiposity in healthy children and adolescents aged 2--15 y? A longitudinal analysis. AB - OBJECTIVE: To investigate the relationship between food energy and macronutrient intake and body fatness assessed up to seven times between 2 and 15 y of age. DESIGN: Prospective, observational study. Generalised linear estimating equations were used to evaluate the longitudinal relationship between body fatness and macronutrient intake. Regression analysis was used to assess whether body fatness at a particular age was predicted by intake at any of the previous ages. SETTING: Community-based project in Adelaide, South Australia. SUBJECTS: In all 143--243 subjects from a representative birth cohort of healthy children recruited in 1975 and followed over 15 y. MAIN OUTCOME MEASURES: The dependent variables were body mass index (BMI), triceps (TC) and subscapular (SS) skinfolds, expressed as standard deviation (s.d.) scores at each age. The predictor variables were energy adjusted macronutrient intake and total energy intake, estimated from a 3--4 day diet diary, the previous corresponding measure of body fatness, sex and parental BMI, TC or SS. RESULTS: Across 2--15 y energy-adjusted fat and carbohydrate intakes were respectively directly and inversely related to SS skinfold measures but not to either BMI or TC skinfold. The best predictor of fatness was previous adiposity, with the effect strengthening as the age interval shortened. Parental BMI, maternal SS and paternal TC contributed to the variance of the corresponding measure in children at some but not all ages. CONCLUSIONS: The current level of body fatness of the child and parental adiposity are more important predictors than dietary intake variables of risk of children becoming or remaining overweight as they grow. PMID- 11423925 TI - Body mass index, weight change and mortality in the elderly. A 15 y longitudinal population study of 70 y olds. AB - OBJECTIVE: To examine the relationship between body mass index (BMI) at age 70, weight change between age 70 and 75, and 15 y mortality. DESIGN: Cohort study of 70-y-olds. SETTING: Geriatric Medicine Department, Goteborg University, Sweden. SUBJECTS: A total of 2628 (1225 males and 1403 females) 70-y-olds examined in 1971--1981 in Gothenburg, Sweden. RESULTS: The relative risks (RRs) for 15 y mortality were highest in the lowest BMI quintiles of males 1.20 (95% CI 0.96- 1.51) and females 1.49 (95% CI 1.14--1.96). In non-smoking males, no significant differences were observed across the quintiles for 5, 10 and 15 y mortality. In non-smoking females, the highest RR (1.58, 95% CI 1.15--2.16) for 15 y mortality was in the lowest quintile. After exclusion of first 5 y death, no excess risks were found in males for following 5 and 10 y mortality across the quintiles. In females, a U-shaped relation was observed after such exclusions. BMI ranges with lowest 15 y mortality were 27--29 and 25--27 kg/m(2) in non-smoking males and females, respectively. A weight loss of > or = 10% between age 70 and 75 meant a significantly higher risk for subsequent 5 and 10 y mortality in both sexes relative to individuals with 'stable' weights. CONCLUSION: Low BMI and weight loss are risk factors for mortality in the elderly and smoking habits did not significantly modify that relationship. The BMI ranges with lowest risks for 15 y mortality are relatively higher in elderly. Exclusion of early deaths from the analysis modified the weight-mortality relationship in elderly males but not in females. PMID- 11423926 TI - Hypophosphataemia in the metabolic syndrome. Gender differences in body weight and blood glucose. AB - OBJECTIVE: We examined gender differences in the clustering of risk factors, especially regarding the possible role of serum phosphate (S-P) in obese patients who participated in a 4 week education programme to reduce their excess risk of cardiovascular disease (CVD). RESULTS: Significantly higher body mass index (BMI) and concentrations of S-P and high density lipids (S-HDL) in serum were found in women (n=1272) than in men (n=993). In women, BMI was inversely correlated with S P at the time of admission. Also in women (but not in men), changes in BMI and S P over 1 or 1-1/2 y were inversely correlated. In men (but not in women), S-P level was inversely correlated with age, systolic blood pressure (SBP), diastolic blood pressure (DBP) and blood glucose at the time of admission. In women, S-P increased and DBP decreased, in addition to weight decrease over both 6 months and 1(1/2) y or 1 y. In women (but not in men), significantly higher age, S-P, S chol, S-TG, S-urate, SBP and DBP values were found with the highest vs the lowest decile of S-Ca. CONCLUSION: We conclude that higher S-P and S-Ca levels may contribute to the difference in metabolic disturbances occurring in obesity, seen in women vs men. PMID- 11423927 TI - Fat-specific satiety in humans for fat high in linoleic acid vs fat high in oleic acid. AB - OBJECTIVE: To investigate the effect of 2 week use of oils high in linoleic (LA), gamma-linolenic (GLA) and oleic acid (OA) on energy intake (EI), macronutrient composition, parameters of appetite and taste perception. DESIGN: A randomized placebo-controlled 2-week treatment, followed by a test day. SETTING: Two-week treatments, daily life; test day, laboratory restaurant. SUBJECTS: Eight overweight men and eight overweight women (body mass index 27.4+/-1.5 kg/m(2)). INTERVENTIONS: Three 2 week treatments, in which subjects replaced their habitual fat products by three different oils (relatively) high in LA (66.8%), GLA (20.2%) or OA (79.6%, placebo). The wash-out periods were 2 weeks. Before each intervention period, taste perception and sensory specific satiety was tested. RESULTS: Even though energy intake was higher during dinner, subjects ate relatively less fat with LA (45.0+/-9.4 E%, P<0.05) than with OA (48.3+/-8.3 E%). Subjects did not distinguish the oils with the different fatty acids from each other. There was no relation between satiety or fat-specific satiety and taste characterization without as well as with sucrose. Although no differences were seen for the AUC of the appetite profile, aet 15:00 h subjects were less satiated with LA (46.1+/-6.2 mm, P<0.05) or GLA (45.1+/-5.8 mm, P<0.01) than after treatment OA (62.5+/-4.8 mm). 24 h EI on the test day was 7.6--8.0 MJ and did not differ between treatments. CONCLUSIONS: Fat-specific satiety during dinner with LA vs OA was shown after a 2-week treatment of each oil, but no change in general satiety. Fat specific satiety was not related to taste perception or characterization of the oils. PMID- 11423928 TI - Hydrogen excretion upon ingestion of dairy products in lactose-intolerant male subjects: importance of the live flora. AB - OBJECTIVE: To assess the effects of the ingestion of milk, yoghurt (10(8) bacteria/ml), heat-treated yoghurt (<15 bacteria/ml) and two products obtained by dilution of yoghurt with heat-treated product (10(6) and 10(5) bacteria/ml) on hydrogen production and symptoms of lactose intolerance in lactose malabsorbers. DESIGN: Double-blind, randomised cross-over design. SETTING: The study was performed in the phase 1 clinical unit of OPTIMED, Nancy, France. SUBJECTS: Twenty-four male lactose malabsorbers were selected for the study. INTERVENTIONS: Hydrogen production and adverse events were followed during 8 h after ingestion of the products. RESULTS: The results clearly demonstrate that ingestion of yoghurt with 10(8) bacteria/ml leads to lower H(2) excretion and complaints than the other products. Results observed with the products containing a reduced population of live flora remain better than those observed with milk. CONCLUSIONS: The importance of a high population of the live flora is underlined. PMID- 11423929 TI - Tissue-specific gene therapy directed to tumor angiogenesis. AB - Gene therapy directed specifically to the vascular wall, particularly to angiogenic endothelial cells is a prerequisite in vascular disease treatment. Angiogenesis is a major feature in many pathological conditions including wound healing, solid tumors, developing metastases, ischemic heart diseases and diabetic retinopathy. In the present study we developed a tissue-specific gene therapy to the angiogenic blood vessels of tumor metastasis using an adeno-based vector containing the murine preproendothelin-1 (PPE-1) promoter. Genes activated by the PPE-1 promoter were highly expressed in bovine aortic endothelial cells in vitro. Systemic injection of the adenoviral vectors AdPPE-1-luciferase and AdCMV luciferase to normal C57BL/6 mice, resulted in higher activity of PPE-1 promoter compared with CMV promoter in the aorta and vascularized tissues such as heart, kidney, lung and pancreas. Systemic administration of the adenoviral vector, in mice bearing Lewis lung carcinoma, resulted in high and specific activity of PPE 1 in the new vasculature of primary tumors and lung metastasis. Cellular distribution of the delivered gene revealed highest expression of GFP in angiogenic endothelial cells of the metastasis. We expect that this approach of 'vascular-directed' gene therapy will be applicable to both vascular diseases and cancer. PMID- 11423930 TI - Mechanism of lipoplex gene delivery in mouse lung: binding and internalization of fluorescent lipid and DNA components. AB - We introduce a lung inflation-fixation protocol to examine the distribution and gene transfer efficiency of fluorescently tagged lipoplexes using fluorescence confocal microscopy within thick lung tissue sections. Using this technique, we tested the hypothesis that factors related to lipoplex distribution were the predominant reason that intravenous (i.v.) administration of lipoplex was superior to intratracheal (i.t.) administration for gene transfer in the murine lung. Lipoplex distribution was analyzed using digitized images of overlapping fields, reconstructed to view an entire lung lobe. Intravenously administered lipoplexes were confined to the capillary network and homogenously distributed throughout the lung lobe. In contrast, i.t. administration resulted in regional distribution of lipoplex, concentrated around bronchioles and distal airways. Not all the bronchioles were stained with lipoplex, suggesting that the airway administered solution became channeled through certain bronchiolar pathways. A fluorescent oligonucleotide was used as a marker for cytoplasmic release of nucleic acids. Quantification of the resulting fluorescent nuclei was used to define the relationship between cytoplasmic release of nucleic acids and gene expression. Endothelial cells were stained after i.v. administration, and epithelial cells were stained after i.t. administration. The delivery of nucleic acids was also more homogeneous with i.v. administration of lipoplex than with i.t. administration. After i.t. administration, it was notable that high concentrations of fluorescent nuclei correlated with low GFP expression. This suggested that toxicity was associated with high local concentrations of cationic lipoplexes. The ratio of GFP-expressing cells to fluorescent nuclei indicated that capillary endothelial cells were more efficient in gene expression per delivery event than were pulmonary epithelial cells. Thus, the greater gene expression efficiency of i.v. administered lipoplexes was due not only to the initial distribution but also to the greater efficiency of the vascular endothelial cells to appropriately traffic and express the foreign gene. PMID- 11423931 TI - Adenovirus-mediated ex vivo gene transfer of basic fibroblast growth factor promotes collateral development in a rabbit model of hind limb ischemia. AB - Adenovirus-mediated ex vivo gene transfer of basic fibroblast growth factor (bFGF), a new strategy for the treatment of chronic vascular occlusive disease, was examined in a rabbit model of hind limb ischemia. The left femoral artery was completely excised to induce an ischemic state in the hind limb of male rabbits. Simultaneously, a skin section was resected from the wound, and host fibroblasts were cultured. The cultured fibroblasts were infected with adenovirus vector containing modified human bFGF cDNA with the secretory signal sequence (AxCAMAssbFGF) or LacZ cDNA (AxCALacZ). At 21 days after femoral artery excision, the gene-transduced fibroblasts were administered through the left internal iliac artery. The fibroblasts significantly accumulated in the ischemic hind limb, and the AxCAMAssbFGF-treated cells secreted bFGF for less than 14 days without elevation of systemic bFGF level. At 28 days after cell administration, calf blood pressure ratio, angiographic score, capillary density of muscle tissue and blood flow of the left internal iliac artery were determined, and animals with AxCAMAssbFGF-treated cells showed significantly greater development of collateral vessels, as compared with those with AxCALacZ-treated cells. These findings suggest that adenovirus-mediated ex vivo gene transfer of bFGF was effective for improvement of chronic limb ischemia. PMID- 11423932 TI - A novel system for the production of fully deleted adenovirus vectors that does not require helper adenovirus. AB - Fully deleted adenovirus vectors (FD-AdVs) would appear to be promising tools for gene therapy. Since these vectors are deleted of all adenoviral genes, they require a helper adenovirus for their propagation. The contamination of the vector preparation by the helper limits the utility of currently existing FD-AdVs in gene therapy applications. We have developed an alternative system for the propagation of FD-AdVs, in which the adenoviral genes essential for replication and packaging of the vector are delivered into producer cells by a baculovirus adenovirus hybrid. A hybrid baculovirus Bac-B4 was constructed to carry a Cre recombinase-excisable copy of the packaging-deficient adenovirus genome. Although the total size of the DNA insert in Bac-B4 was 38 kb, the genetic structure of this recombinant baculovirus was stable. Bac-B4 gave high yields in Sf9 insect cells, with titers of 5 x 10(8)p.f.u./ml before concentration. Transfection of 293-Cre cells with lacZ-expressing FD-AdV plasmid DNA followed by infection by Bac-B4 at a MOI of 2000 p.f.u./ml resulted in rescue of the helper-free vector. Subsequent passaging of the obtained FD-AdV using Bac-B4 as a helper resulted in approximately 100-fold increases of the vector titer at each passage. This resulting vector was completely free of helper virus and was able to transduce cultured 293 cells. However, scaling-up of FD-AdV production was prevented by the eventual emergence of replication-competent adenovirus (RCA). Experiments are underway to optimize this system for the large-scale production of helper virus free FD-AdVs and to minimize the possibility of generation of replication competent adenovirus (RCA) during vector production. This baculovirus-based system will be a very useful alternative to current methods for the production of FD-AdVs. PMID- 11423933 TI - Synthesis of a novel series of cationic lipids that can act as efficient gene delivery vehicles through systematic heterocyclic substitution of cholesterol derivatives. AB - The synthesis of a series of novel cationic lipids through the systematic substitution of cholesterol derivatives that could greatly enhance the delivery and expression of plasmid DNA in vitro and in vivo is described. Two of the newly synthesized lipids, designated as NCC4 and NCC10, were chosen to be studied in detail and gave much higher levels of gene expression than that which could be obtained with some of the conventional cationic polymers and cationic liposomes. In vivo studies with both NCC4 and NCC10 also showed better ability in delivering the reporter gene to the target cells through intrasplenic injection. In addition, by varying the DNA/lipid charge ratios, NCC4 and NCC10 can withstand serum inactivation in vitro. However, this does not correlate with the corresponding increase in the level of gene expression following systemic gene delivery with NCC4 and NCC10 in vivo. PMID- 11423934 TI - Recombinant AAV vectors containing the foot and mouth disease virus 2A sequence confer efficient bicistronic gene expression in cultured cells and rat substantia nigra neurons. AB - Recombinant adeno-associated viruses (rAAVs) are promising vectors for gene therapy since they efficiently and stably transduce a variety of tissues of immunocompetent animals. The major disadvantage of rAAVs is their limited capacity to package foreign DNA (< or =5 kb). Often, co-expression of two or more genes from a single viral vector is desirable to achieve maximal therapeutic efficacy or to track transduced cells in vivo by suitable reporter genes. The internal ribosome entry site (IRES) sequence of encephalomyocarditis virus has been widely used to construct bicistronic viral vectors. However, the IRES is rather long and IRES-mediated translation can be relatively inefficient when compared with cap-dependent translation. As an alternative to the IRES for in vivo gene expression, we studied the 16 amino-acid long 2A peptide of foot and mouth disease virus (FMDV). The 2A peptide mediates the primary cis-'cleavage' of the FMDV polyprotein in a cascade of processing events that ultimately generate the mature FMDV proteins. We have generated several different rAAV genomes in which two coding regions are fused in-frame via the FMDV 2A sequence. We show that FMDV 2A efficiently mediates the generation of the expected cleavage products from the artificial fusion proteins in cells. Furthermore, we find that both EGFP and alpha- synuclein are expressed at substantially higher levels from 2A vectors than from the corresponding IRES-based vectors, while SOD-1 is expressed at comparable or slightly higher levels. Finally, we demonstrate for the first time, that the 2A sequence results in effective bicistronic gene expression in vivo after injection of 2A-dependent rAAVs into the rat substantia nigra. We conclude that 2A-containing rAAVs may represent an attractive alternative to IRES-dependent vectors for ex vivo and in vivo gene expression and gene therapy. PMID- 11423935 TI - Inhibition of hepadnaviral replication by polyethylenimine-based intravenous delivery of antisense phosphodiester oligodeoxynucleotides to the liver. AB - Antisense oligodeoxynucleotides (ODNs) appear as attractive anti-hepatitis B virus (HBV) agents. We investigated in vivo, in the duck HBV (DHBV) infection model, whether linear polyethylenimine (lPEI)-based intravenous delivery of the natural antisense phosphodiester ODNs (O-ODNs) can prevent their degradation and allow viral replication inhibition in the liver. DHBV-infected Pekin ducklings were injected with antisense O-ODNs covering the initiation codon of the DHBV large envelope protein, either in free form (O-ODN-AS2) or coupled to lPEI (lPEI/O-ODN-AS2). Following optimization of lPEI/O-ODN complex formulation, complete O-ODN condensation into a homogenous population of small (20-60 nm) spherical particles was achieved. Flow cytometry analysis showed that lPEI mediated transfer allowed the intrahepatic delivery of lPEI/O-ODN-AS2 to increase three-fold as compared with the O-ODN-AS2. Following 9-day therapy the intrahepatic levels of both DHBV DNA and RNA were significantly decreased in the lPEI/O-ODN-AS2-treated group as compared with the O-ODN-AS2-treated, control lPEI/O-ODN-treated, and untreated controls. In addition, inhibition of intrahepatic viral replication by lPEI/O-ODN-AS2 was not associated with toxicity and was comparable with that induced by the phosphorothioate S-ODN-AS2 at a five fold higher dose. Taken together, our results demonstrate that phosphodiester antisense lPEI/O-ODN complexes specifically inhibit hepadnaviral replication. Therefore we provide here the first in vivo evidence that intravenous treatment with antisense phosphodiester ODNs coupled to lPEI can selectively block a viral disease-causing gene in the liver. PMID- 11423936 TI - Adenovirus-mediated transfer of type IV collagen alpha5 chain cDNA into swine kidney in vivo: deposition of the protein into the glomerular basement membrane. AB - Gene therapy of Alport syndrome (hereditary nephritis) aims at the transfer of a corrected type IV collagen alpha chain gene into renal glomerular cells responsible for production of the glomerular basement membrane (GBM). A GBM network composed of type IV collagen molecules is abnormal in Alport syndrome which leads progressively to kidney failure. The most common X-linked form of the disease is caused by mutations in the gene for the alpha5(IV) chain, the alpha5 chain of type IV collagen. Full-length human alpha5(IV) cDNA was expressed in HT1080 cells with an adenovirus vector, and the recombinant alpha5(IV) chain was shown to assemble into heterotrimers consisting of alpha3(IV) and alpha4(IV) chains, utilizing a FLAG epitope in the recombinant alpha5(IV) chain. The results indicate that correction of the molecular defect in Alport syndrome is possible. Previously, we had developed an organ perfusion method for effective in vivo gene transfer into glomerular cells. In vivo perfusion of pig kidneys with the recombinant adenovirus resulted in expression of the alpha5(IV) chain in kidney glomeruli as shown by in situ hybridization and its deposition into the GBM was shown by immunohistochemistry. The results strongly suggest future possibilities for gene therapy of Alport syndrome. PMID- 11423937 TI - T cells engrafted with a recombinant anti-CD30 receptor target autologous CD30(+) cutaneous lymphoma cells. AB - T cells can be directed to antigen-specific, MHC-independent target cell lysis by grafting with a recombinant receptor with antibody-like specificity. Here, we asked whether T cells from the peripheral blood of a patient with cutaneous T cell lymphoma can be recruited for an immune response against autologous tumor cells. Lymphoma cells with a CD3(-) CD4(+) CD30(+) phenotype and clonal TCR Vbeta7 rearrangement were isolated from a cutaneous lesion. The lymphoma lesion additionally harbored CD3(+) CD25(+) activated normal T cells despite ongoing tumor progression. Peripheral blood-derived T cells from the lymphoma patient were retrovirally engrafted with a recombinant anti-CD30-scFv-gamma receptor. Upon cocultivation with autologous CD30(+)lymphoma cells, grafted T cells increase IFN-gamma secretion and lyse specifically lymphoma cells with high efficiency, even at an effector to target cell ratio of as low as 1:20. Our data demonstrate that the recombinant anti-CD30-gamma receptor overcomes T cell tolerance for tumor cells and directs T cells specifically against autologous lymphoma cells. PMID- 11423938 TI - Preface to special issue on Sphingomonas. PMID- 11423939 TI - Chlorophenol and nitrophenol metabolism by Sphingomonas sp UG30. AB - Sphingomonas sp UG30 is a pentachlorophenol (PCP)-degrading bacterial strain capable of degrading several nitrophenolic compounds, including p-nitrophenol (PNP), 2,4-dinitrophenol (2,4-DNP), p-nitrocatechol and 4,6-dinitro-o-cresol (DNOC). The ability to degrade both chlorophenolic and nitrophenolic compounds is probably not restricted to UG30, but may also be possessed by other pentachlorophenol-degrading Sphingomonas spp. The interesting question arises as to whether there is any point of convergence between the initial pathways of PCP and nitrophenol degradation in these microorganisms. There is some experimental evidence that PCP-4-monooxygenase is involved in metabolism of both p-nitrophenol and 2,4-dinitrophenol. Further studies are needed to confirm this and to examine the role(s) of other PCP-degrading enzymes in nitrophenol metabolism by this microorganism. In this paper, we review some of the taxonomic, biochemical, physiological and ecological properties of Sphingomonas sp UG30 with respect to biodegradation of PCP and nitrophenolic compounds. PMID- 11423940 TI - Chemotaxonomic characterisation of Sphingomonas. AB - Based on published results and investigations done for this study, chemotaxonomic characteristics of all validly described species of the genus Sphingomonas, as well as unnamed strains of this genus and related genera such as Rhizomonas and Blastomonas, are presented. All representatives of this group, here designated as sphingomonads, contain ubiquinone (Q-10). The two different polyamine patterns characterized either by the predominant polyamine sym-homospermidine or spermidine separate the sphingomonads into two major groups. Complex polar lipid profiles were found in sphingomonads in addition to the characteristic compound sphingoglycolipid. Identical profiles were found only in a few phylogenetically highly related species. Common to all sphingomonads is a fatty acid composition with 2-hydroxy fatty acids (14:0 2OH in all currently recognized species) and the lack of 3-hydroxy acids, which distinguishes them from taxa outside this group. Qualitative and quantitative differences in the fatty acid compositions, in several cases, were also suitable for identification at the species level. Thus, the differences in the chemotaxonomic characteristics demonstrate that the analyses of these low molecular weight cell compounds are suitable for classification of sphingomonads. PMID- 11423941 TI - Detection of Sphingomonas spp in soil by PCR and sphingolipid biomarker analysis. AB - Sphingomonas spp possess unique abilities to degrade refractory contaminants and are found ubiquitously in the environment. We developed Sphingomonas genus specific PCR primers (SPf-190 and SPr1-852) which showed specific amplification of a 627-bp 16S rDNA fragment from Sphingomonas spp. A PCR assay using these Sphingomonas specific primers was developed to detect Sphingomonas aromaticivorans B0695R in three texturally distinct soil types, showing detection limits between 1.3-2.2 x 10(3) CFU g(-1) dry soil. A sphingolipid extraction protocol was also developed to monitor Sphingomonas populations in soil quantitatively. The detection limit of the assay was 20 pmol g(-1) dry soil, equivalent to about 3 x 10(5) cells g(-1) dry soil. Survival of S. aromaticivorans B0695R was monitored in the three different soils by antibiotic selective plate counting, PCR and sphingolipid analysis. All three approaches showed that the B0695R cells persisted in the low biomass Sequatchie sub-soil at about 3-5 x 10(7)cells g(-1) dry soil. In comparison to the plate counting assay, both the PCR and sphingolipid analysis detected a significantly higher level of B0695R cells in the clay soil and Sequatchie top-soil, indicating the possibility of the presence of viable but non-culturable B0695R cells in the soils. The combination of PCR and sphingolipid analysis may provide a more realistic estimation of Sphingomonas population in the environment. PMID- 11423942 TI - Detection of sphingomonads and in situ identification in activated sludge using 16S rRNA-targeted oligonucleotide probes. AB - The increasing significance of members of the genus Sphingomonas in biotechnological applications has led to an increased interest in the diversity, abundance and ecophysiological potential of this group of Gram-negative bacteria. This general focus provides a challenge to improve means for identification of sphingomonads; eg molecular genetic methods for rapid and specific detection could facilitate screening of new isolates. Here, fluorescently labeled oligonucleotide probes targeted against 16S rRNA were used to typify strains previously assigned to the genus. All 46 sphingomonads tested including type strains of 21 Sphingomonasspecies could be detected with a probe originally designed for the genus and all but one with a probe designed for the alpha-4 subgroup of the Proteobacteria. The two probes are suitable for direct detection of sphingomonads in pure and mixed cultures as well as in environmental samples of unknown composition. The probes were used to identify sphingomonads in situ in activated sludge samples. Sphingomonads were rather abundant accounting for about 5-10% of the total cells in municipal sludges. Distinct patterns in aggregation of the cells suggest that these organisms could be involved in the formation process of sludge flocs. PMID- 11423943 TI - Sphingomonads from marine environments. AB - Sphingomonas species play an important role in the ecology of a range of marine habitats. Isolates and 16S-rRNA clones have been obtained from corals, natural and artificial sources of marine hydrocarbons and eutrophic and oligotrophic waters, and have been isolated as hosts for marine phages. In addition they are found in oceans spanning temperature ranges from polar to temperate waters. While less is known about marine sphingomonads in comparison to their terrestrial counterparts, their importance in microbial ecology is evident. This is illustrated by, for example, the numerical dominance of strain RB2256 in oligotrophic waters. Furthermore, the known marine sphingomonads represent a phylogenetic cross-section of the Sphingomonas genus. This review focuses on our present knowledge of cultured isolates and 16S-rDNA clones from marine environments. PMID- 11423944 TI - Ecology, physiology, and phylogeny of deep subsurface Sphingomonas sp. AB - Several new species of the genus Sphingomonas including S. aromaticivorans, S. stygia, and S. subterranea that have the capacity for degrading a broad range of aromatic compounds including toluene, naphthalene, xylenes, p-cresol, fluorene, biphenyl, and dibenzothiophene, were isolated from deeply-buried (>200 m) sediments of the US Atlantic coastal plain (ACP). In S. aromaticivorans F199, many of the genes involved in the catabolism of these aromatic compounds are encoded on a 184-kb conjugative plasmid; some of the genes involved in aromatic catabolism are plasmid-encoded in the other strains as well. Members of the genus Sphingomonas were common among aerobic heterotrophic bacteria cultured from ACP sediments and have been detected in deep subsurface environments elsewhere. The major source of organic carbon for heterotrophic metabolism in ACP deep aquifers is lignite that originated from plant material buried with the sediments. We speculate that the ability of the subsurface Sphingomonas strains to degrade a wide array of aromatic compounds represents an adaptation for utilization of sedimentary lignite. These and related subsurface Sphingomonas spp may play an important role in the transformation of sedimentary organic carbon in the aerobic and microaerobic regions of the deep aquifers of the ACP. PMID- 11423945 TI - Beijerinckia sp strain B1: a strain by any other name. AB - Beijerinckia sp strain B1 grows with biphenyl as its sole source of carbon and energy. A mutant, strain B8/36, oxidized biphenyl to cis-(2S,3R)-dihydroxy-l phenylcyclohexa-4,6-diene (cis-biphenyl dihydrodiol). Strain B8/36 oxidized anthracene, phenanthrene, benz[a]anthracene and benzo[a]pyrene to cis dihydrodiols. Other substrates oxidized to cis-dihydrodiols were dibenzofuran, dibenzothiophene and dibenzo-p-dioxin. Biphenyl dioxygenase activity was observed in cells of Beijerinckia B1 and B8/36 after growth in the presence of biphenyl, m , p-xylene and salicylate. Recent studies have led to the reclassification of Beijerinckia B1 as Sphingomonas yanoikuyae strain B1. Subsequent biotransformation studies showed that S. yanoikuyae B8/36 oxidized chrysene to a bis-cis-diol with hydroxyl substituents at the 3,4- and 9,10-positions. Dihydronaphthalene was oxidized to cis-1,2-dihydroxy-1,2,3,4 tetrahydronaphthalene, naphthalene, cis-1,2-dihydroxy-1,2-dihydronaphthalene and 2-hydroxy-1,2-dihydronaphthalene. Anisole and phenetole were oxidized to phenol. Thus the S. yanoikuyae biphenyl dioxygenase catalyzes cis-dihydroxylation, benzylic monohydroxylation, desaturation and dealkylation reactions. To date, the genes encoding biphenyl dioxygenase have not been cloned. However, the nucleotide sequence of a S. yanoikuyaeB1 DNA fragment contains five different alpha subunits as determined by conserved amino acids coordinating iron in a Rieske [2Fe-2S] center and mononuclear iron at the catalytic site. The specific role of the different putative oxygenases in biotransformation reactions catalyzed by S. yanoikuyae is not known and presents an exciting challenge for future studies. PMID- 11423946 TI - Functional analysis of genes involved in biphenyl, naphthalene, phenanthrene, and m-xylene degradation by Sphingomonas yanoikuyae B1. AB - Sphingomonas yanoikuyae B1 is able to utilize toluene, m-xylene, p-xylene, biphenyl, naphthalene, phenanthrene, and anthracene as sole sources of carbon and energy for growth. A forty kilobase region of DNA containing most of the genes for the degradation of these aromatic compounds was previously cloned and sequenced. Insertional inactivation of bphC results in the inability of B1 to grow on both polycyclic and monocyclic compounds. Complementation experiments indicate that the metabolic block is actually due to a polar effect on the expression of bphA3, coding for a ferredoxin component of a dioxygenase. Lack of the ferredoxin results in a nonfunctional polycyclic aromatic hydrocarbon dioxygenase and a nonfunctional toluate dioxygenase indicating that the electron transfer components are capable of interacting with multiple oxygenase components. Insertional inactivation of a gene for a dioxygenase oxygenase component downstream of bphA3 had no apparent effect on growth besides a polar effect on nahD which is only needed for growth of B1 on naphthalene. Insertional inactivation of either xylE or xylG in the meta-cleavage operon results in a polar effect on bphB, the last gene in the operon. However, insertional inactivation of xylX at the beginning of this cluster of genes does not result in a polar effect suggesting that the genes for the meta-cleavage pathway, although colinear, are organized in at least two operons. These experiments confirm the biological role of several genes involved in metabolism of aromatic compounds by S. yanoikuyae B1 and demonstrate the interdependency of the metabolic pathways for polycyclic and monocyclic aromatic hydrocarbon degradation. PMID- 11423947 TI - Induction of aromatic catabolic activity in Sphingomonas aromaticivorans strain F199. AB - Enzyme induction studies with Sphingomonas aromaticivorans F199 demonstrated that both toluene and naphthalene induced expression of both naphthalene and toluene catabolic enzymes. However, neither aromatic compound induced expression of all the enzymes required for complete mineralization of either naphthalene or toluene. Activity measurements in combination with gene sequence analyses indicate that growth on either aromatic substrate in the absence of the other is, therefore, sub-optimal and is predicted to lead to the build-up of metabolites due to imbalance in toluene or naphthalene catabolic enzyme activities. Growth on toluene may be further inhibited by the co-expression of two toluene catabolic pathways, as predicted from gene sequence analyses. One of these pathways may potentially result in the formation of a dead-end intermediate, possibly benzaldehyde. In contrast, either p-cresol or benzoate can support high levels of growth. Analyses of promoter region sequences on the F199 aromatic catabolic plasmid, pNL1, suggest that additional regulatory events are modulated through the interaction of BphR with Sigma54 type promoters and through the binding of a regulator upstream of p-cresol catabolic genes and xylM. We hypothesize that the unusual gene clustering in strain F199 is optimized for simultaneous degradation of multiple aromatic compound classes, possibly in response to the heterogeneous composition of aromatic structures in the fossil organic matter present in the deep Atlantic Coastal Plain sediments from which this bacterium was isolated. PMID- 11423948 TI - Sphingomonas paucimobilis BPSI-3 mutant AN2 produces a red catabolite during biphenyl degradation. AB - The biphenyl degradation pathway of Sphingomonas paucimobilis BPSI-3 was investigated using a degradation-deficient mutant generated by 1-methyl-3-nitro-1 nitrosoguanidine (NTG) mutagenesis. The mutant, designated AN2, was confirmed as originating from BPSI-3 through the use of ERIC (Enterobacterial Repetitive Intergenic Consensus) PCR and by detection of the diagnostic pigment, nostoxanthin, in cellular methanol extracts. Mutant AN2 produced a yellow followed by red extracellular substance when grown in the presence of biphenyl. In the presence of 2,3-dihydroxybiphenyl, yellow followed by red then yellow compounds were formed over time. This colour change was consistent with the characteristics of a quinone, 1-phenyl-2,3-benzoquinone, which could arise from the oxidation of 2,3-dihydroxybiphenyl. A quinone was synthesised from 2,3 dihydroxybiphenyl and compared to the red compound produced by mutant AN2. Gas chromatography-mass spectrophotometry (GC-MS) confirmed that a similar quinone (4,5-dimethoxy-3-phenyl-1,2-benzoquinone) compared to the structure of the proposed biogenic compound, had been formed. This compound was also found after GC-MS analysis of mutant AN2 culture extracts. Spectrophotometric analysis of the quinone synthesised and the red product produced revealed almost identical spectral profiles. A likely inference from this evidence is that the mutant AN2 is blocked, or its activity altered, in the first gene cluster, bphA to C, of the biphenyl degradation pathway. PMID- 11423949 TI - Phylogeny of Sphingomonas species that degrade pentachlorophenol. AB - Four pentachlorophenol (PCP)-degrading bacteria isolated from geographically diverse areas have been examined in detail as regards their physiology and phylogeny. According to traditional biochemical methods, these strains had been classified as members of the genera Arthrobacter, Flavobacterium, Pseudomonas, and Sphingomonas. The PCP degradation pathway has been studied extensively in Sphingomonas (Flavobacterium) sp strain ATCC 39723 and the first three degradation steps catalyzed by a PCP-4-monooxygenase (PcpB) and a reductive dehalogenase (PcpC) that functions twice are well established. A fourth step appears to involve ring-fission of the aromatic nucleus (PcpA). Molecular analyses revealed that the PCP degradation pathway in these four strains was rather conserved, leading to a phylogenetic analysis using 16S rDNA. The results revealed a much closer phylogenetic relationship between these organisms than traditional classification indicated, placing them into the more recently established genus Sphingomonas where they may even represent a single species. With 16S rDNA analysis, many bacterial isolates involved in degradation of xenobiotic compounds that were previously classified into diverse genera have been reclassified into the genus Sphingomonas. PMID- 11423950 TI - Colonization of contaminated soil by an introduced bacterium: effects of initial pentachlorophenol levels on the survival of Sphingomonas chlorophenolica strain RA2. AB - The survival of a Sphingomonas species that was introduced into pentachlorophenol (PCP)-contaminated soil was monitored with two complementary methods, a respiration-based assay and a most probable number (MPN) technique. Sphingomonas chlorophenolicastrain RA2 is a PCP-mineralizing bacterium that was introduced into soil contaminated with a range of PCP concentrations (0-300 &mgr;g PCP g(-1) soil). The population of introduced microorganisms was followed for 170 days using a substrate-induced growth-response method and a MPN assay that specifically targets PCP-mineralizing bacteria. Varying the initial PCP concentration resulted in the emergence of three distinct patterns of survival. In soil contaminated with 300 &mgr;g PCP g(-1) the population of S. chlorophenolica strain RA2 immediately declined following introduction, increased by 200-fold and leveled off by the end of the 170-day incubation. In contrast, populations of S. chlorophenolica strain RA2 declined to levels below detection limits in uncontaminated soil by the end of the experiment. Intermediate PCP concentrations (10-100 &mgr;g PCP g(-1) soil) resulted in the establishment of S. chlorophenolica strain RA2 that slowly declined in numbers. These results indicate that Sphingomonas chlorophenolica strain RA2 is an effective colonizer of PCP-contaminated soil but will not persist in the absence of PCP. PMID- 11423951 TI - Degradation of the phenoxy acid herbicide diclofop-methyl by Sphingomonas paucimobilis isolated from a Canadian prairie soil. AB - Sphingomonas paucimobilis, isolated from a soil in Manitoba, Canada, was able to utilize diclofop-methyl, (R,S)-methyl-2-[4-(2,4 dichlorophenoxy)phenoxy]propionate, as the sole source of carbon and energy. An actively growing aerobic culture completely degraded 1.5 &mgr;g diclofop-methyl ml(-1) to diclofop acid within 54 h, at 25 degrees C. A biphasic growth pattern indicated that this organism was capable of degrading diclofop acid to 4-(2,4 dichlorophenoxy)phenol and 2,4-dichlorophenol and/or phenol. The accumulation of 2,4-dichlorophenol in the growth medium, however, suggested that Sphingomonas paucimobilis was unable to utilize this compound as a source of carbon and energy. PMID- 11423952 TI - Sphingomonas herbicidovorans MH: a versatile phenoxyalkanoic acid herbicide degrader. AB - Sphingomonas herbicidovorans MH was isolated from a dichlorprop-degrading soil column. It is able to grow on phenoxyalkanoic acid herbicides, such as mecoprop, dichlorprop, 2,4-D, MCPA, and 2,4-DB. Strain MH utilizes both enantiomers of the chiral herbicides mecoprop and dichlorprop as sole carbon and energy sources. Enantiomer-specific uptake systems are responsible for transporting the acidic substrates across the cell membrane. Catabolism is initiated by two enantiomer specific alpha-ketoglutarate-dependent dioxygenases that catalyze the cleavage of the ether bond of the respective enantiomer to yield the corresponding phenol and pyruvate. Therefore selective degradation of the enantiomers of mecoprop and dichlorprop by strain MH is not only due to enantioselective catabolism but also to enantioselective transport. PMID- 11423953 TI - Degradation of azo dyes containing aminonaphthol by Sphingomonas sp strain 1CX. AB - Sphingomonas sp strain 1CX was isolated from a wastewater treatment plant and is capable of aerobically degrading a suite of azo dyes, using them as a sole source of carbon and nitrogen. All azo dyes known to be decolorized by strain 1CX (Orange II, Acid Orange 8, Acid Orange 10, Acid Red 4, and Acid Red 88) have in their structure either 1-amino-2-naphthol or 2-amino-1-naphthol. In addition, an analysis of the structures of the dyes degraded suggests that there are certain positions and types of substituents on the azo dye which determine if degradation will occur. Growth and dye decolorization occurs only aerobically and does not occur under fermentative or denitrification conditions. The mechanism by which 1CX decolorizes azo dyes appears to be through reductive cleavage of the azo bond. In the case of Orange II, the initial degradation products were sulfanilic acid and 1-amino-2-naphthol. Sulfanilic acid, however, was not used by 1CX as a growth substrate. The addition of glucose or inorganic nitrogen inhibited growth and decoloration of azo dyes by 1CX. Attempts to grow the organism on chemically defined media containing several different amino acids and sugars as sources of nitrogen and carbon were not successful. Phylogenetic analysis of Sphingomonas sp strain 1CX shows it to be related to, but distinct from, other azo dye decolorizing Sphingomonas spp strains isolated previously from the same wastewater treatment facility. PMID- 11423954 TI - 5-Chloropicolinic acid is produced by specific degradation of 4-chlorobenzoic acid by Sphingomonas paucimobilis BPSI-3. AB - We have previously shown that the bacterium Sphingomonas paucimobilis BPSI-3, isolated from PCB-contaminated soil, can degrade halogenated biphenyls, naphthalenes, catechols and benzoic acids. However, before such an organism can be used in bioremediation, it is important to characterise the degradation products and determine the degradation pathways to ensure that compounds more toxic or mobile than the original contaminants are not produced. In the degradation of 4-chlorobiphenyl, S. paucimobilis BPSI-3 produces a novel chlorinated picolinic acid. In this paper, we show that 4-chlorobenzoate is an intermediate in this degradation and, through (15)N-labelling, that 5 chloropicolinate is the only nitrogenous metabolite isolated under the extraction conditions used. The position of the chlorine indicates that degradation of 4 chlorocatechol occurs exclusively via a 2,3-extradiol cleavage. These data allow us to postulate a more definitive catabolic pathway for the biodegradation of 4 chlorobiphenyl to 5-chloro-2-hydroxymuconic acid semialdehyde via 4 chlorobenzoate in S. paucimobilis BPSI-3. PMID- 11423955 TI - Interaction of Sphingomonas and Pseudomonas strains in the degradation of chlorinated dibenzofurans. AB - We have studied the concerted degradation of two monochlorodibenzofurans by a bacterial consortium, consisting of the chlorodibenzofurans-cometabolizing and chlorosalicylates-excreting strain Sphingomonas sp RW16, and Pseudomonas sp RW10, which mineralized the released chlorosalicylates. Neither of the organisms was able to grow with chlorodibenzofurans alone. Degradation of 2-chloro- and 3 chlorodibenzofuran proceeded to the end products 5-chloro- and 4 chlorosalicylate, respectively, when the initial dioxygenase of Sphingomonas sp RW 16 attacked the unchlorinated aromatic ring of the heterocyclic dibenzofuran molecule. 2-Hydroxypenta-2,4-dienoate, formed upon meta-cleavage of the intermediary chlorotrihydroxybiphenyls, served as a growth substrate for the sphingomonad. Presumably, most of the chlorosalicylates were excreted and degraded further by Pseudomonas sp RW10. Mineralization of both chlorosalicylates proceeded through a converging pathway, via 4-chlorocatechol, and protoanemonin. Chlorosalicylates were mineralized by the pseudomonad only when their concentration in the culture medium was below 1.5 mM. In the case of initial dioxygenation taking place on the chlorinated aromatic ring, salicylate and chlorinated hydroxypentadienoates should be formed. The metabolic fate of putative chlorohydroxypentadienoates is not clear; ie, they may be channeled into unproductive catabolism and, thus, represent the critical point in the breakdown of the carbon of these two chlorodibenzofurans by Sphingomonas sp RW16. PMID- 11423956 TI - Catabolism of 2,7-dichloro- and 2,4,8-trichlorodibenzofuran by Sphingomonas sp strain RW1. AB - Due to their physicochemical and toxicological properties, polychlorinated dibenzofurans are regarded as a class of compounds providing reason for serious environmental concern. While the nonhalogenated basic structure dibenzofuran is effectively mineralized by appropriate bacterial strains, its polychlorinated derivatives are not. To elucidate the ability of the strain Sphingomonas sp RW1 to metabolize some of these chlorinated derivatives, we performed turnover experiments using 2,7-dichloro- and 2,4,8-trichlorodibenzofuran. As indicated by the oxygen-uptake rates determined for these two chlorinated dibenzofurans, Sphingomonassp RW1 can catabolize these chlorinated dibenzofurans yielding small quantities of oxidation products, which we isolated and subsequently characterized employing GC/MS and (1)H- as well as (13)C-NMR spectroscopy. In the case of 2,7-dichlorodibenzofuran, two metabolites accumulated, which we identified as 6-chloro- and 7-chloro-2-methyl-4H-chromen-4-one. The single metabolite isolated from the turnover experiments performed with 2,4,8 trichlorodibenzofuran was unequivocally identified as 6,8-dichloro-2-methyl-4H chromen-4-one. PMID- 11423957 TI - Characterization of Sphingomonas paucimobilis SYK-6 genes involved in degradation of lignin-related compounds. AB - Sphingomonas paucimobilis SYK-6 is able to grow on a wide variety of dimeric lignin compounds. These compounds are degraded via vanillate and syringate by a unique enzymatic system, composed of etherases, O demethylases, ring cleavage oxygenases and side chain cleaving enzymes. These unique and specific lignin modification enzymes are thought to be powerful tools for utilization of the most abundant aromatic biomass, lignin. Here, we focus on the genes and enzymes involved in beta-aryl ether cleavage and biphenyl degradation. Two unique etherases are involved in the reductive cleavage of beta-aryl ether. These two etherases have amino acid sequence similarity with the glutathione S transferases, and use glutathione as a hydrogen donor. It was found that 5,5' dehydrodivanillate, which is a typical lignin-related biphenyl structure, was transformed into 5-carboxyvanillate by the reaction sequence of O-demethylation, meta-ring cleavage, and hydrolysis, and the genes involved in the latter two reactions have been characterized. Vanillate and syringate are the most common intermediate metabolites in lignin catabolism. These compounds are initially O demethylated and the resulting diol compounds, protocatechuate (PCA) and 3-O methylgallate, respectively, are subjected to ring cleavage catalyzed by PCA 4,5 dioxygenase. The ring cleavage products generated are further degraded through the PCA 4,5-cleavage pathway. We have isolated and characterized genes for enzymes involved in this pathway. Disruption of a gene for 2-pyrone-4,6 dicarboxylate hydrolase (ligI) in this pathway suggested that an alternative route for 3-O-methylgallate degradation, in which ligI is not involved, would play a role in syringate catabolism. In this article, we describe the genetic and biochemical features of the S. paucimobilis SYK-6 genes involved in degradation of lignin-related compounds. A possible application of the SYK-6 lignin degradation system to produce a valuable chemical material is also described. PMID- 11423958 TI - Lessons learned from Sphingomonas species that degrade abietane triterpenoids. AB - Abietane terpenoid-degrading organisms include Sphingomonas spp which inhabit natural environments and biological treatment systems. An isolate from the high Arctic indicates that these organisms occur far from trees which synthesize abietanes and suggests that some of these organisms can occupy a niche in hydrocarbon-degrading soil communities. Abietane-degrading Sphingomonas spp provide additional evidence that the phylogeny of this genus is independent of the catabolic capabilities of its members. Studies of Sphingomonas sp DhA-33 demonstrate that biological treatment systems for pulp mill effluents have the potential to mineralize abietane resin acids. On the other hand, these studies indicate that some chlorinated dehydroabietic acids are quite recalcitrant. Strain DhA-33 grows relatively well on some chlorinated dehydroabietic acids but transforms others to stable metabolites. Using strain DhA-33, a novel method was developed to measure the metabolic activity of an individual population within a complex microbial community. Oligonucleotide hybridization probes were used to assay the 16S rRNA:rDNA ratio of DhA-33 as it grew in an activated sludge community. However, this method proved not to be sufficiently sensitive to measure naturally occurring resin acid-degrading populations. We propose that the same approach can be modified to use more sensitive assays. PMID- 11423959 TI - Complete analysis of genes and enzymes for gamma-hexachlorocyclohexane degradation in Sphingomonas paucimobilis UT26. AB - gamma-Hexachlorocyclohexane (gamma-HCH; also called BHC or lindane) is one of the highly chlorinated pesticides which can cause serious environmental problems. Sphingomonas paucimobilis UT26 degrades gamma-HCH under aerobic conditions. The unique degradation pathway of gamma-HCH in UT26 is revealed. In the upstream pathway, gamma-HCH is transformed to 2,5-dichlorohydroquinone (2,5-DCHQ) by two different dehalogenases (LinA and LinB) and one dehydrogenase (LinC) which are expressed constitutively. In the downstream pathway, 2,5-DCHQ is reductively dehalogenated, and then ring-cleaved by enzymes (LinD and LinE, respectively) whose expressions are regulated. We have cloned and sequenced five structural genes (linA, linB, linC, linD, and linE) directly involved in this degradation pathway. The linD and linE genes form an operon, and its expression is positively regulated by the LysR-type transcriptional regulator (LinR). The genes linA, linB, and linC are constitutively expressed, and are present separately from each other in the UT26 genome. Cell fractionation analysis, Western blotting, and immuno electron microscopy revealed that LinA and LinB are localized in the periplasmic space of UT26. PMID- 11423960 TI - Degradation of substituted naphthalenesulfonic acids by Sphingomonas xenophaga BN6. AB - Sphingomonas xenophaga BN6 was isolated from the river Elbe as a member of a multispecies bacterial culture which mineralized 6-aminonaphthalene-2-sulfonate. Pure cultures of strain BN6 converted a wide range of amino- and hydroxynaphthalene-2-sulfonates via a catabolic pathway similar to that described for the metabolism of naphthalene to salicylate by Pseudomonas putida NAH7 or Pseudomonas sp NCIB 9816. In contrast to the naphthalene-degrading pseudomonads, S. xenophaga BN6 only partially degraded the naphthalenesulfonates and excreted the resulting amino- and hydroxysalicylates in almost stoichiometric amounts. Enzymes that take part in the degradative pathway of the naphthalenesulfonates by strain BN6 were purified, characterized and compared with the isofunctional enzymes from the naphthalene-degrading pseudomonads. According to the enzyme structures and the catalytic constants, no fundamental differences were found between the 1,2-dihydroxynaphthalene dioxygenase or the 2'-hydroxybenzalpyruvate aldolase from strain BN6 and the isofunctional enzymes from the naphthalene degrading pseudomonads. The limited available sequence information about the enzymes from strain BN6 suggests that they show about 40-60% sequence identity to the isofunctional enzymes from the pseudomonads. In addition to the gene for the 1,2-dihydroxynaphthalene dioxygenase, the genes for two other extradiol dioxygenases were cloned and sequenced from strain BN6 and the corresponding gene products were studied. S. xenophaga BN6 has also been used as a model organism to study the mechanism of the non-specific reduction of azo dyes under anaerobic conditions and to establish combined anaerobic/aerobic treatment systems for the degradation of sulfonated azo dyes. Furthermore, the degradation of substituted naphthalenesulfonates by mixed cultures containing strain BN6 was studied in continuous cultures and was described by mathematical models. PMID- 11423961 TI - Sphingomonads involved in the biodegradation of xenobiotic polymers. AB - Sphingomonads involved in the microbial degradation of xenobiotic polymers are introduced. The metabolism of polyethylene glycol was the primary focus of the study. Several others, including polyvinyl alcohol, polyethylene and polyaspartate were also studied. It is suggested that these xenobiotic polymers are metabolized by intracellular enzymes located in the periplasmic space or bound to membranes, indicating that transport of these polymers through outer membranes is requisite for their metabolism. Involvement of specific membrane structures of sphingomonads such as unusual sphingolipids is suggested for membrane transport of xenobiotic compounds, especially hydrophobic materials. PMID- 11423962 TI - Chemical structure and function of glycosphingolipids of Sphingomonas spp and their distribution among members of the alpha-4 subclass of Proteobacteria. AB - Sphingomonas spp are phylogenetically placed in the alpha-4 subclass of Proteobacteria. They have glycosphingolipids (GSL) in their membranes instead of lipopolysaccharide (LPS) as in other Gram-negative bacteria. S. paucimobilis, the type species of the genus, has GSL-1, which contains only glucuronic acid (GlcA) as a sugar moiety, and GSL-4A, which contains a tetrasaccharide including GlcA. GSL-1 and GSL-4A form the outer membrane of S. paucimobilis with outer membrane proteins and phospholipids. In the outer membrane, GSLs are assumed to locate and function as does the LPS of other Gram-negative bacteria. Sphingomonas spp closely related to the type species contain both GSL-1 and the oligosaccharide type GSL such as GSL-4A, but other Sphingomonas spp and other genera in the alpha 4 subclass of Proteobacteria contain only GSL-1. Structural variations of fatty acids and dihydrosphingosines in the GSL-1 are presented. PMID- 11423963 TI - Interaction of peptides and proteins with bacterial surface glycolipids: a comparison of glycosphingolipids and lipopolysaccharides. AB - The bacterial cell wall of Gram-negative bacteria consists, in addition to the cytoplasmic membrane, of another permeability barrier, the outer membrane. The lipid distribution between both sides of this membrane is strictly asymmetric. The outer leaflet is made up of glycolipids, usually lipopolysaccharides. In Sphingomonas spp glycosphingolipids were found to substitute for lipopolysaccharides. In this review, it is shown by an electrophysiological approach that glycosphingolipid can replace lipopolysaccharide with respect to its function as antigenic surface structure as well as to its contribution to the diffusion barrier properties of the outer membrane. This review is focused on: (i) the function of porins, as examples of transmembrane proteins, in the different glycolipid environments; (ii) the interaction of polymyxin B with the outer membrane, as an example of polycationic antibacterial peptides; and (iii) the activation of the human complement system by lipopolysaccharides and glycosphingolipids. PMID- 11423964 TI - Special cell surface structure, and novel macromolecule transport/depolymerization system of Sphingomonas sp A1. AB - A bacterium isolated from soil as an alginate lyase producer shows characteristic morphological and taxonomical properties consistent with being classified in the genus Sphingomonas. The bacterium utilizes high molecular weight (HMW)-alginate for growth by depolymerization of the polymer with intracellular alginate lyases, which are generated from a common precursor protein through autoregulated post translational modifications. Electron microscopic observations of the cell surface and of thin sections of cells grown on HMW-alginate revealed dynamic changes in both cell surface and membrane structures. The most remarkable change is recognized in the formation of mouth-like pits which open and close depending on the presence or absence of HMW-alginate. Enzymatic and genetic analyses of HMW alginate incorporation processes confirmed the presence of a pit-dependent and macromolecule-specific ABC transporter system in cells of Sphingomonas species A1. This is the first description of a bacterium with a pit on the cell surface and a pit-dependent endocytosic uptake system for macromolecules. PMID- 11423965 TI - Planktonic/sessile dimorphism of polysaccharide-encapsulated sphingomonads. AB - Sphingomonads have acquired diverse metabolic activities to inhabit a wide range of environments. Several strains of Sphingomonas display phenotypic dimorphism and can adopt either a planktonic or sessile behavior in liquid media. The sessile state is marked by the presence of a viscous exopolysaccharide capsule. Specific types of these capsular polysaccharides are harvested from large-scale fermentations for use as rheology modifiers in many industrial and food applications. Sensing of environmental stimuli and genetic control over synthesis of the capsule are key events in alternating between these two phenotypes. PMID- 11423966 TI - Evaluation of rheological properties of the exopolysaccharide of Sphingomonas paucimobilis GS-1 for application in oil exploration. AB - Analysis of an exopolysaccharide of Sphingomonas paucimobilis GS-1 (EPS/GS-1) with respect to its rheological properties, cross-linking ability with chrome alum and performance test at 75 +/- 5 degrees C revealed its strong suspending ability, shear thinning property, and thixotrophic nature which are required to impart desirable rheology to drilling mud. The organism fulfilled all the specified requirements and its properties were superior to those of currently used XC polymer (a xanthan product) for oil drilling applications. However, EPS/GS-1 was unstable in the presence of bentonite at 100 +/- 5 degrees C during performance tests, in contrast to XC polymer. PMID- 11423967 TI - Direct regulation of the centrosome duplication cycle by the p53-p21Waf1/Cip1 pathway. AB - The function of the centrosomes to direct mitotic spindles is critical for accurate chromosome transmission to daughter cells. Since each daughter cell inherits one centrosome, each centrosome must duplicate prior to the next mitosis, and do so only once. Thus, there are control mechanism(s) that ensure the coordinated progression of centrosome duplication and other cell cycle events (i.e. DNA synthesis), and limit centrosome duplication to once per cell cycle. Deregulation of the centrosome duplication cycle results in abnormal amplification of centrosomes, leading to aberrant mitoses and increased chromosome transmission errors. This has been found to be the case for cells lacking functional p53 tumor suppressor protein. However, it had remained to be determined whether the deregulation of the centrosome duplication cycle is the direct or indirect effect of loss/mutational inactivation of p53. Here, we found that the normal centrosome duplication cycle is almost completely restored in p53(-/-) cells by re-introduction of wild-type p53 at a physiologically relevant level, demonstrating that p53 is directly involved in the regulation of centrosome duplication. Since cyclin dependent kinase 2 (CDK2)/cyclin E triggers DNA synthesis as well as centrosome duplication, we tested whether Waf1, a CDK inhibitor and a major target of p53's transactivation function, is an effector of p53-mediated regulation of centrosome duplication. We found that induced expression of Waf1 in p53(-/-) cells only partially restored the centrosome duplication control, suggesting that Waf1 comprises one of the multiple effector pathways of the p53-mediated regulation of the centrosome duplication cycle. PMID- 11423968 TI - Regulation of microtubule assembly by human EB1 family proteins. AB - The EB1 family proteins are highly conserved microtubule-associated proteins. The EB1 protein in yeast has been shown to play an important role in regulating microtubule dynamics and chromosome segregation. Human EB1 family proteins include EB1, RP1 and EBF3. Although EB1 and RP1 have been shown to associate with microtubules, the subcellular localization of endogenous EBF3 had not been characterized. The function of human EB1 family proteins was also not clear. We therefore investigated the cellular localization of EBF3 and the regulation of microtubule organization by EB1 family proteins. As do EB1 and RP1, EBF3 was found to colocalize with microtubules, preferentially at their plus ends, throughout the cell cycle. Moreover, there was a very strong EBF3 signal at the centrosome in interphase cells and at the spindle poles in mitotic cells. When EB1 family proteins were overexpressed, they associated with the entire microtubule cytoskeleton. In addition, EB1 and EBF3 induced microtubule bundling in some cells overexpressing these proteins. These microtubule bundles were more resistant to nocodazole and were more acetylated than regular microtubules. Our results demonstrate for the first time that human EB1 family proteins could regulate microtubule assembly and stability. PMID- 11423969 TI - p63alpha and DeltaNp63alpha can induce cell cycle arrest and apoptosis and differentially regulate p53 target genes. AB - The p53 tumor suppressor protein plays a critical role in the regulation of the cell cycle and apoptosis. The importance of p53's functions is underscored by the high incidence of p53 mutations in human cancers. Recently, two p53-related proteins, p73 and p63, were identified as members of the p53 gene family. Multiple isoforms of p73 have been found, including DeltaN variants in which the N-termini are truncated. p63 is expressed as three major forms, p63alpha, p63beta and p63gamma, each of which differ in their C-termini. All three forms can be alternatively transcribed from a cryptic promoter located within intron 3, producing DeltaNp63alpha, DeltaNp63beta and DeltaNp63gamma. The high degree of similarity of p73 and p63 to evolutionarily conserved regions of p53 suggests that these proteins play an important and potentially redundant role in regulating cell cycle arrest and apoptosis. Here we describe the characterization of cell lines generated to inducibly express p63alpha and DeltaNp63alpha. We have found that p63alpha and DeltaNp63alpha can differentially regulate endogenous p53 target genes and induce cell cycle arrest and apoptosis. Deletion of the N terminal 26 amino acids of DeltaNp63alpha abolished its ability to transactivate p53 target genes and induce cell cycle arrest and apoptosis. This indicates that a putative transactivation domain exists within the N-terminus of the DeltaN variants of p63. Furthermore, the differential regulation of p53 target genes by p63alpha and DeltaNp63alpha suggests that p63 and p53 utilize both similar and different signaling pathways to execute their cellular functions. PMID- 11423970 TI - Activation of p53 by roscovitine-mediated suppression of MDM2 expression. AB - The p53 tumor suppressor is regulated by the MDM2 oncoprotein. Overexpression of MDM2 maintains p53 at low levels and contributes to the functional inactivation of p53 in a subset of tumors. We found that treatment with roscovitine and olomoucin, which were originally developed as cyclin-dependent kinase (CDK) inhibitors, can efficiently stabilize and activate nuclear p53 in tumor cells with MDM2 amplification or cytoplasmic p53. These inhibitors block the degradation of p53 without affecting p53-MDM2 binding and the nuclear shuttling function of p53 and MDM2. Roscovitine also induces stabilization of the p53 Ala 315 mutant, indicating that it does not act by regulating the CDK phosphorylation of serine 315. Roscovitine induces down-regulation of MDM2 expression at both protein and mRNA levels. Ectopic expression of MDM2 can abrogate the ability of roscovitine to induce p53 stabilization. Low concentrations of roscovitine cooperate with the DNA-damaging agent camptothecin to activate p53 in a synergistic fashion. These results show that the small molecule CDK inhibitors can be used to activate p53 through their potent inhibitory effect on MDM2 expression and may be useful as sensitizing agents for other DNA-damaging drugs. PMID- 11423971 TI - The Gem GTP-binding protein promotes morphological differentiation in neuroblastoma. AB - Gem is a small GTP-binding protein within the Ras superfamily whose function has not been determined. We report here that ectopic Gem expression is sufficient to stimulate cell flattening and neurite extension in N1E-115 and SH-SY5Y neuroblastoma cells, suggesting a role for Gem in cytoskeletal rearrangement and/or morphological differentiation of neurons. Consistent with this potential function, in clinical samples of neuroblastoma, Gem protein was most highly expressed within cells which had differentiated to express ganglionic morphology. Gem was also observed in developing trigeminal nerve ganglia in 12.5 day mouse embryos, demonstrating that Gem expression is a property of normal ganglionic development. Although Gem expression is rare in epithelial and hematopoietic cancer cell lines, constitutive Gem levels were detected in several neuroblastoma cell lines and could be further induced as much as 10-fold following treatment with PMA or the acetylcholine muscarinic agonist, carbachol. PMID- 11423972 TI - Blocked B cell differentiation and emigration support the early growth of Myc induced lymphomas. AB - Avian leukosis virus induces lymphoma in chickens after proviral integration within the c-Myc gene, and subsequent expansion of Myc-overexpressing lymphocytes within transformed bursal follicles. The clonal expansion of these follicles allowed us to examine how Myc influences cell differentiation, growth, and apoptosis in lymphoid progenitors soon after the onset of Myc overexpression. Immunohistochemical analysis of developmental markers established that Myc overexpression consistently blocks lymphocyte differentiation at a late embryonic stage. Myc-transformed follicles also grow much more rapidly than normal follicles. This rapid growth is not mediated by suppression of apoptosis, as normal and Myc-transformed follicles showed similar rates of cell death by TUNEL immunohistochemical analysis of cells undergoing DNA degradation. Measurements of DNA synthesis and mitotic index showed modest effects of Myc to increase lymphocyte proliferation, as normal lymphocytes already divide rapidly. The major mechanism mediating rapid growth of transformed follicles instead involved failure of myc-overexpressing lymphocytes to emigrate from transformed follicles, while normal lymphocytes actively emigrate after hatching, as measured by BrdU pulse-chase labeling and immunohistochemical measurements. This failure to undergo the normal program of differentiation and subsequent bursal retention of lymphocytes accounts for most of the growth of transformed follicles, while Myc induced proliferation makes a smaller contribution. PMID- 11423973 TI - Altered gene expression in immunogenic poorly differentiated thyroid carcinomas from RET/PTC3p53-/- mice. AB - Cancers develop and progress via activation of oncogenes and loss of tumor suppressor genes, a progression that can be recapitulated through cross breeding mouse strains harboring genetic mutations. To define the role of RET/PTC3, p53 and Fhit in thyroid carcinogenesis, we intercrossed RET/PTC3 transgenics with p53 /- mice. This new strain, RET/PTC3p53-/-, succumb to rapidly growing and strikingly large multilobed thyroid tumors containing mixtures of both well and poorly differentiated, highly proliferative follicular epithelial cells. Interestingly, transplanted tumors from RET/PTC3p53-/- mice grew in SCID but not syngeneic immunocompetent mice indicating that these advanced tumors were immunogenic. RET/PTC3 protein expression was reduced to undetectable levels in tumors of older mice suggesting that the continued elevated expression of RET/PTC3 may not be necessary for tumor progression. Similarly, expression of Fhit protein was reduced in early tumors and undetected in older tumors irrespective of tumor histopathology. In contrast to RET/PTC3p53-/- mice, RET/PTC3Fhit-/- mice did not develop advanced thyroid carcinomas. These studies support a model of human thyroid cancer whereby thyroid epithelium expresses RET/PTC3 protein at early stages of tumor development, followed by the reduction of RET/PTC3 and loss of p53 function with progressive reduction of Fhit protein expression coincident with malignant progression. PMID- 11423974 TI - Protein kinase CK2 in mammary gland tumorigenesis. AB - Protein kinase CK2 is a ubiquitous and evolutionarily conserved serine/threonine kinase that is upregulated in many human cancers and can serve as an oncogene in lymphocytes. Recently, we have demonstrated that CK2 potentiates Wnt/beta-catenin signaling in mammary epithelial cells. To determine whether CK2 overexpression contributes to mammary tumorigenesis, we have performed comparative studies of human and rat breast cancer specimens and we have engineered transgenic mice with dysregulated expression of CK2alpha in the mammary gland. We find that CK2 is highly expressed in human breast tumor specimens and in carcinogen-induced rat mammary tumors. Overexpression of CK2alpha in the mammary gland of transgenic mice, under control of the MMTV-LTR, causes hyperplasia and dysplasia of the female mammary gland. Thirty per cent of the female MMTV-CK2alpha transgenic mice develop mammary adenocarcinomas at a median of 23 months of age, often associated with Wnt pathway activation, as evidenced by upregulation of beta-catenin protein. NF-kappaB activation and upregulation of c-Myc also occur frequently. Thus, in mice, rats, and humans, dysregulated expression of CK2 is associated with and is capable of contributing to mammary tumorigenesis. Targeted inhibition of CK2 could be useful in the treatment of breast cancer. PMID- 11423975 TI - Analysis of the expression of cell cycle regulators in Ewing cell lines: EWS-FLI 1 modulates p57KIP2and c-Myc expression. AB - Ewing tumour is characterized by specific chromosome translocations which fuse EWS to a subset of genes encoding ETS transcription factors, most frequently FLI 1. We report the analysis of the expression of various cell cycle regulators both in Ewing tumour derived cell lines and in different cellular models with either inducible or constitutive EWS-FLI-1 cDNA expression. In Ewing cell lines, cyclin D1, CDK4, Rb, p27KIP1 and c-Myc were consistently highly expressed whereas p57KIP2, p15INK4B and p14ARF demonstrated undetectable or low expression levels. The amount of p16INK4A, p21CIP1, p18INKAC and CDK6 was variable from one cell line to the other. The inducible expression of EWS-FLI-1 led to a strong upregulation of c-Myc and a considerable downregulation of p57KIP2. Other proteins did not show evident modification. High c-Myc and very low p57KIP2 expression levels were also observed in neuroblastoma NGP cells constitutively expressing EWS-FLI-1 as compared to parental cells. Analysis of the p57KIP2 promoter indicated that EWS-FLI-1 downregulates, possibly through an indirect mechanism, the transcription of this gene. Finally, we show that ectopic expression of p57KIP2 in Ewing cells blocks proliferation through a complete G1 arrest. These results suggest that the modulation of p57(KIP2) expression by EWS FLI-1 is a fundamental step in Ewing tumorigenesis. PMID- 11423976 TI - Ionizing radiation modulates vascular endothelial growth factor (VEGF) expression through multiple mitogen activated protein kinase dependent pathways. AB - We investigated the role of radiation-induced mitogen activated protein kinase (MAPK) pathway activity in the regulation of proliferation, cell survival and vascular endothelial growth factor (VEGF) production in primary astrocytes and in T9 and RT2 glioblastoma cells derived from Fisher 344 rats. In these cells, ionizing radiation (2 Gy) caused activation of the MAPK pathway which was blocked by specific inhibitor drugs. Blunting of radiation-induced MAPK activity weakly enhanced radiation-induced apoptosis 24 h after exposure in RT2 cells. Furthermore, blunting of MAPK activation weakly enhanced the ability of radiation to reduce RT2 cell growth in clonogenic growth assays. These findings argue that inhibition of MAPK signaling reduces proliferation and enhances cell killing by ionizing radiation in transformed astrocytes. Proliferation and survival of cancer cells has been linked in vivo to enhanced expression of angiogenic growth factors. Recently we demonstrated that the gene product of a novel rodent radiation-responsive gene, progression elevated gene 3 (PEG-3), could enhance vascular endothelial growth factor (VEGF) promoter activity in rodent fibroblasts, leading to increased VEGF protein levels and tumorigenic behavior in vivo. Thus PEG-3 and VEGF expression could be expected to directly correlate with the oncogenic potential of transformed cells. RT2 cells expressed more PEG-3 and VEGF protein than T9 cells, and were more tumorigenic in vivo than T9 cells. Radiation activated the PEG-3 promoter via MAPK signaling and ectopic over expression of PEG-3 enhanced both basal MAPK activity and basal VEGF promoter activity. Basal MAPK activity partially correlated with basal VEGF promoter activity and VEGF protein levels in primary astrocytes, T9 and RT2 cells. Radiation increased the activity of the VEGF promoter and VEGF protein levels in primary astrocytes, T9 and RT2 cells which were dependent upon MAPK function. Furthermore, inhibition of AP-1 transcription factor signaling by dominant negative c-Jun (TAM67) also significantly reduced basal, and to a lesser extent radiation-induced, VEGF promoter function in RT2 cells. Collectively, our data demonstrate that radiation-induced MAPK signaling can both protect cells from radiation-induced cell death as well as enhance protein levels of pro-angiogenic factors such as VEGF. Enhanced VEGF expression in RT2 cells may be mediated via MAPK and JNK pathway signaling which converges upon the AP-1 transcription factor complex. PMID- 11423977 TI - The synovial sarcoma associated protein SYT interacts with the acute leukemia associated protein AF10. AB - As a result of the synovial sarcoma associated t(X;18) translocation, the human SYT gene on chromosome 18 is fused to either the SSX1 or the SSX2 gene on the X chromosome. Although preliminary evidence indicates that the (fusion) proteins encoded by these genes may play a role in transcriptional regulation, little is known about their exact function. We set out to isolate interacting proteins through yeast two hybrid screening of a human cDNA library using SYT as a bait. Of the positive clones isolated, two were found to correspond to the acute leukemia t(10;11) associated AF10 gene, a fusion partner of MLL. Confirmation of these results was obtained via co-immunoprecipitation of endogenous and exogenous, epitope-tagged, SYT and AF10 proteins from cell line extracts and colocalization of epitope-tagged SYT and AF10 proteins in transfected cells. Subsequent sequential mutation analysis revealed a highly specific interaction of N-terminal SYT fragments with C-terminal AF10 fragments. The N-terminal interaction domain of the SYT protein was also found to be present in several SYT orthologs and homologs. The C-terminal interaction domain of AF10 is located outside known functional domains. Based on these results, a model is proposed in which the SYT and AF10 proteins act in concert as bipartite transcription factors. This model has implications for the molecular mechanisms underlying the development of both human synovial sarcomas and acute leukemias. PMID- 11423978 TI - Cyclin G1 is involved in G2/M arrest in response to DNA damage and in growth control after damage recovery. AB - Cyclin G1 is one of the target genes of the transcription factor p53, and is induced in a p53-dependent manner in response to DNA damage. Although cyclin G1 has been implicated in a range of biological phenomena, its precise function remains unclear. Here we present an analysis of the physiological role of cyclin G1 using mice homozygous for a targeted disruption of the cyclin G1 gene. In order to clarify the role of cyclin G1 in the p53 pathway, downstream events such as apoptosis, cell growth and cell cycle checkpoint control were analysed in thymocytes and embryonic fibroblasts derived from cyclin G1-disrupted mice. No difference was detected in induction of apoptosis between mouse embryo fibroblasts (MEFs) derived from cyclin G1+/+ and cyclin G1-/- mice. Following irradiation, cyclin G1-/- MEFs proliferated more slowly and reached lower cell densities in culture dishes than cyclin G1+/+ MEFs. Analysis of cell survival showed that cyclin G1-/- MEFs were about twice as sensitive as cyclin G1+/+ MEFs to gamma radiation or UV radiation. Cyclin G1-/- mice were more sensitive to gamma radiation than wild-type mice. Flow cytometeric analysis revealed that the number of cyclin G1-/- MEFs in G2/M phase after irradiation was reduced by 50% relative to cyclin G1+/+ MEFs. Our results demonstrate that cyclin G1 plays roles in G2/M arrest, damage recovery and growth promotion after cellular stress. PMID- 11423979 TI - Mutations in the mitotic check point gene, MAD1L1, in human cancers. AB - Aneuploidy is a characteristic of the majority of human cancers, and recent studies suggest that defects of mitotic checkpoints play a role in carcinogenesis. MAD1L1 is a checkpoint gene, and its dysfunction is associated with chromosomal instability. Rare mutations of this gene have been reported in colon and lung cancers. We examined a total of 44 cell lines (hematopoietic, prostate, osteosarcoma, breast, glioblastoma and lung) and 133 fresh cancer cells (hematopoietic, prostate, breast and glioblastoma) for alterations of MAD1L1 by RT-PCR-SSCP and nucleotide sequencing. Eight mutations consisting of missense, nonsense and frameshift mutations were found, together with a number of nucleotide polymorphisms. All the alterations in cell lines were heterozygous. Frequency of mutations was relatively high in prostate cancer (2/7 cell lines and 2/33 tumor specimens). We placed a mutant truncated MAD1L1, found in a lymphoma sample, into HOS, Ht161 and SJSA cell lines and found that it was less inhibitory than wild type MAD1L1 at decreasing cell proliferation. Co-expression experiments showed that the mutant form had a dominant-negative effect. Furthermore, this mutant impaired the mitotic checkpoint as shown by decreased mitotic indices in HOS cells expressing mutant MAD1L1 after culture with the microtubule-disrupting agent, nocodazole. Our results suggest a pathogenic role of MAD1L1 mutations in various types of human cancer. PMID- 11423980 TI - Post-transcriptional inactivation of p53 in immortalized murine embryo fibroblast cells. AB - The steady-state levels of p53 mRNA and protein were barely detectable by Northern and Western blot analysis in spontaneously immortalized (10)3 and (10)7 murine embryo fibroblast (MEF) cells. But when cells were treated with cycloheximide (CHX) or emetine, expression levels were restored to those observed in primary and immortal (10)10 MEF cells. However, levels of p53 mRNA were not changed in primary or (10)10 MEF cells by CHX treatment. De novo p53 mRNA synthetic rates were similar in primary, (10)10, (10)3, and (10)7 MEF cells treated with or without CHX. Treatment with actinomycin D (ActD) showed that p53 mRNA in primary and (10)10 MEF cells had a relatively long half-life of 22 h, compared to less than 2 h for (10)3 and (10)7 MEF cells. Pulse-chase analysis of p53 mRNA turnover using CHX and ActD showed that the rapid destabilization of p53 mRNA in (10)3 and (10)7 MEF cells could be regulated at the transcriptional and translational levels. In addition, the destabilization of p53 mRNA appeared to occur in the nucleus for (10)3 and (10)7 cells, but not for primary and (10)10 MEF cells. Taken together, the present study demonstrates that inactivation of the p53 gene occurs at the post-transcriptional level by rapid destabilization of its mRNA in the nucleus of spontaneously immortalized (10)3 and (10)7 MEF cells. PMID- 11423981 TI - Novel properties of the cyclin encoded by Human Herpesvirus 8 that facilitate exit from quiescence. AB - Viral DNA replication is generally dependent upon circumventing host cell cycle control to force S phase entry in an otherwise quiescent cell. Here we describe novel attributes of the cyclin encoded by Human Herpesvirus 8 (K cyclin) that enable it to subvert the quiescent state. K cyclin is most similar to the mammalian D-type cyclins in primary sequence but displays properties more akin to those of cyclin E. K cyclin (like cyclin E) can autonomously couple with its cognate cdk subunit and localize to the nucleus. D-type cyclins require mitogen stimulated accessory factors (such as p21(Cip1) and p27(Kip1)) to facilitate both of these processes. A striking difference between K cyclin and mammalian cyclins is that K cyclin binding to cdk6 can substantially activate the catalytic activity of the complex without the requirement for cyclin H/cdk7 phosphorylation of the cdk T-loop; this phosphorylation is obligatory for endogenous cyclin/cdk activity. However, K cyclin/cdk6 complexes are not totally immune from cell cycle control since CAK phosphorylation is necessary for complete activation. Thus, CAK phosphorylated K cyclin/cdk6 targets multiple sites in the retinoblastoma protein (pRb) whereas the unphosphorylated complex targets a single site. The restricted substrate specificity of the non-CAK phosphorylated K cyclin/cdk6 complex is insufficient to enable K cyclin-mediated S phase entry. Thus, the viral K cyclin is reliant upon endogenous CAK activity to subvert the quiescent state. PMID- 11423982 TI - The hepatitis B virus HBx protein induces adherens junction disruption in a src dependent manner. AB - Chronic hepatitis B virus infection is strongly associated with the development of hepatocellular carcinoma (HCC). Epithelial tumors are frequently characterized by loss of cadherin expression or function. Cadherin-dependent adhesion prevents the acquisition of a migratory and invasive phenotype, and loss of its function is itself enough for the progression from adenoma to carcinoma. The HBx protein of hepatitis B virus is thought to contribute to the development of the carcinoma, however, its role in the oncogenic and metastatic processes is far from being fully understood. We report herein the ability of HBx to disrupt intercellular adhesion in three different cell lines stably transfected with an inducible HBx expression vector. The linkage between the actin cytoskeleton and cadherin complex, which is essential for its function, is disrupted in the presence of HBx, as indicated by detergent solubility and immunoprecipitation experiments. In addition, beta-catenin was tyrosine phosphorylated in HBx expressing cells. Inhibition of the src family of tyrosine kinases resulted in the prevention of the disruption of adherens junctions. These results suggest that HBx is able to disrupt intercellular adhesion in a src-dependent manner, and provide a novel mechanism by which HBx may contribute to the development of HCC. PMID- 11423983 TI - Smad3/AP-1 interactions control transcriptional responses to TGF-beta in a promoter-specific manner. AB - Smad proteins transduce signals from TGF-beta receptors and regulate transcription of target genes either directly or in combination with other sequence-specific transcription factors. AP-1 sites and their cognate transcription factors also play important roles in the gene regulatory activities of TGF-beta. In this report, we have investigated the functional interactions of the Smad and AP-1 transcription factors. We demonstrate that Smad and AP-1 complexes specifically bind to their cognate cis-elements and do not interact with each other on-DNA, whereas off-DNA interactions occur between Smad3 and both c-Jun and JunB. Using both artificial constructs specific for either the Smad or AP-1 signaling pathways or natural promoters known to be TGF-beta-responsive, we have determined that Jun family members downregulate Smad3-mediated gene transactivation whereas AP-1-dependent promoters are synergistically activated by Smad3 and Jun proteins. We propose a model where the presence of Smad- and/or AP 1-specific cis-elements within TGF-beta-responsive genes allows dynamic modulation of gene expression, in contrast to the existing model where interactions between Smad and AP-1 proteins are merely an on/off mechanism to regulate TGF-beta/Smad targets. PMID- 11423984 TI - Distinct pattern of p53 phosphorylation in human tumors. AB - The protein product of the tumor suppressor gene p53 is phosphorylated on multiple residues by several protein kinases. Using a battery of 10 antibodies developed against different phosphorylated and acetylated residues of p53, we compared the pattern of p53 phosphorylation and acetylation in tumor-derived cell lines, tumor samples, and non-neoplastic cells. Irrespective of tumor types or the presence of p53 mutation, phosphorylation and acetylation of p53 was substantially higher in samples obtained from tumor tissues than those found in non-transformed samples. Among the 10 sites analysed, phosphorylation of residues 15, 81, 392, and acetylation were among the more frequent modifications. Analysis of two of the more abundant phosphorylation or acetylation sites on p53 is sufficient to detect 72% of tumor-derived p53 proteins. The distinct pattern of p53 phosphorylation and acetylation in human tumors may offer a new means to monitor the status and activity of p53 in the course of tumor development and progression. PMID- 11423985 TI - Hypermethylation of 14-3-3 sigma (stratifin) is an early event in breast cancer. AB - We have identified 14-3-3 sigma (sigma) as a gene whose expression is lost in breast carcinomas, primarily by methylation-mediated silencing. In this report, we investigated the timing of loss of sigma gene expression during breast tumorigenesis in vivo. We analysed the methylation status of sigma in breast cancer precursor lesions using microdissection for selective tissue sampling. We found hypermethylation of sigma in 24 of 25 carcinomas (96%), 15 of 18 (83%) of ductal carcinoma in situ, and three of eight (38%) of atypical hyperplasias. None of the five hyperplasias without atypia showed sigma-hypermethylation. Unexpectedly, patients with breast cancer showed sigma hypermethylation in adjacent histologically normal breast epithelium, while this was never observed in individuals without evidence of breast cancer. Also, samples of periductal stromal breast tissue were consistently hypermethylated, underscoring the importance of selective tissue sampling for accurate assessment of 14-3-3-sigma methylation in breast epithelium. These results suggest that hypermethylation of 14-3-3-sigma occurs at an early stage in the progression to invasive breast cancer, and may occur in apparently normal epithelium adjacent to breast cancer. These results provide evidence that loss of expression of sigma is an early event in neoplastic transformation. PMID- 11423986 TI - Staurosporine induces apoptosis through both caspase-dependent and caspase independent mechanisms. AB - Sensitivity of tumor cells to anticancer therapy depends on the ability of the drug to induce apoptosis. However, multiple signaling pathways control this induction and thus determine this sensitivity. We report here that staurosporine, a well known inducer of apoptosis in a wide range of cell lines, displays distinct ability to trigger apoptosis in two different L1210 sublines (termed L1210/S and L1210/0). Staurosporine treatment resulted in an early cell death (within 3 h) in L1210/S cells, while in L1210/0 cells, death occurred only after 12 h. In both instances, death occurred by apoptosis. A broad spectrum caspase inhibitor, Z-VAD-fmk, blocked early apoptosis in L1210/S cells but did not confer any protection on late apoptosis in L1210/0 cells. Protection by Z-VAD-fmk observed in L1210/S cells was not lasting and unmasked a secondary process of cell death that also exhibited characteristics of apoptosis. Thus, staurosporine induces apoptotic cell death through at least two redundant parallel pathways. These two pathways normally coexist in L1210/S cells. However, the early cell death mechanism depending on caspase activation disguises the late caspase independent apoptotic process. Staurosporine-induced apoptosis in L1210/0 cells develops only by the caspase-independent mechanism due to a general defect in caspase activation. PMID- 11423987 TI - Dominant-negative transcription factor AP-2 augments SB-2 melanoma tumor growth in vivo. AB - We have previously demonstrated that the transition of melanoma to the metastatic phenotype is associated with a loss of expression of the transcription factor AP 2. To further investigate the role of AP-2 in the progression of human melanoma, we attempted to inactivate AP-2 in primary cutaneous SB-2 melanoma cells by using a dominant-negative AP-2, or AP-2B, gene. AP-2B is an alternatively spliced AP-2 variant capable of inhibiting AP-2 trans-activator function. Stable transfection of primary cutaneous melanoma SB-2 cells with the dominant-negative AP-2B gene was confirmed by RT--PCR and Northern blot analyses. Electromobility shift assay using nuclear extracts from these cell lines demonstrated decreased functional binding of AP-2B-transfected cells to the AP-2 consensus binding sequence compared with neo-transfected controls. In addition, CAT activity driven by a construct containing the AP-2 consensus binding sequence was downregulated in the AP-2B transfected cells, indicating AP-2 activity was quenched in the transfected cells. Orthotopic (subcutaneous) injection of the dominant-negative (AP-2B) transfected cell lines into nude mice increased their tumorigenicity compared to control neo-transfected cells. The AP-2B-transfected cells displayed an increase in MMP-2 expression (by Northern blot) and MMP-2 activity (by zymography), which resulted in an increase in invasiveness through Matrigel-coated filters. The AP 2B-transfected tumors also displayed an increase in MMP-2 expression, microvessel density, and angiogenesis in vivo. These results demonstrate that inactivation of AP-2 contributes to the progression of melanoma, at least partially via deregulation of the MMP-2 gene. PMID- 11423988 TI - Inhibition of cyclin A/Cdk2 phosphorylation impairs B-Myb transactivation function without affecting interactions with DNA or the CBP coactivator. AB - Expression of the B-Myb transcription factor is directed by an E2F-dependent transcriptional mechanism to late G1 and S phases of the cell cycle, where its transactivation properties are enhanced post-translationally by cyclin A/Cdk2 mediated phosphorylation. Other experiments have shown that removal of the B-Myb C-terminus constitutively activates both transactivation and DNA-binding activities, suggesting that autoregulation by this inhibitory domain is counteracted by phosphorylation. We report here on further experiments to examine this hypothesis. The importance of this modification was first emphasized by showing that co-transfected dominant-negative Cdk2 (Cdk2DN) substantially reduced B-Myb transactivation activity. We then attempted to map the autoregulatory domain by analysing a series of progressively deleted C-terminal B-Myb mutants. Removal of just 29 C-terminal aa increased transactivation appreciably, however, maximal activity required removal of 143 amino acids (as in B-Myb + 561). Enhanced B-Myb + 561 function correlated with the acquisition of DNA binding activity to a single Myb binding site (MBS) oligonucleotide as determined by bandshift assays, however, further assays showed that even wt B-Myb could bind a DNA fragment containing three MBS. Although transactivation by B-Myb was severely dependent on hyperphosphorylation, neither inhibiting this activity by co transfecting Cdk2DN nor augmenting it with cyclin A resulted in significant effects on DNA-binding. We also found that B-Myb could synergize with the CBP coactivator and that this cooperativity was cyclin A/Cdk2-dependent. Despite this, the physical association between these proteins was not influenced by the B Myb phosphorylation status. We discuss these findings in relation to the autoregulation of B-Myb by the C-terminal domain. PMID- 11423989 TI - p21 Waf1/Cip1 can protect human colon carcinoma cells against p53-dependent and p53-independent apoptosis induced by natural chemopreventive and therapeutic agents. AB - The molecular basis for the sensitivity of tumor cells to chemopreventive natural food compounds and commonly used chemotherapeutic agents is not well understood, not least because studies are frequently confounded by the diversity among cell lines or rely on experimental protein overexpression. Here we investigated the effects of n-butyrate, a cancer-preventive short-chain fatty acid produced by anaerobic bacteria in the gastrointestinal tract, on the human wild-type p53 and p21 expressing HCT116 colon carcinoma cell line and on HCT116 cells with either p53 or p21 alleles inactivated by homologous recombination. The effects of n butyrate were then compared with those elicited by cytotoxic drugs and the natural chemopreventive phytoalexin of wine and grapes, resveratrol. We document that physiological concentrations of n-butyrate stimulate p21 expression and induce apoptosis independently of p53, and that the absence of p21 increases apoptosis drastically. The apoptosis is mediated through the mitochondria and is accompanied by mitochondrial proliferation and membrane potential changes. Adriamycin, etoposide, cisplatinum, colcemid and resveratrol induce distinct cellular responses; however, absence of p21 favors apoptosis-induction by adriamycin, etoposide and colcemid. Thus, control of p21 expression may support chemoprevention and certain tumor therapies. PMID- 11423990 TI - Soluble CD44 inhibits melanoma tumor growth by blocking cell surface CD44 binding to hyaluronic acid. AB - Proteolytic cleavage of the extracellular domain of CD44 from the surface of cells has been observed recently in different cell types. In cell culture supernatants of human melanoma cell lines a 70 kDa soluble CD44 protein (solCD44) was detected at concentrations of 250-300 ng/ml. Protease inhibitor studies revealed that serine proteases and metalloproteases are involved in the cleavage of CD44 from the surface of melanoma cells. To analyse a possible function of soluble CD44 a human malignant melanoma cell line was stably transfected with cDNAs encoding either wild type soluble CD44s or mutated forms with defective HA binding properties (CD44sR41A and CD44sR150A/R154A). Soluble CD44s almost completely inhibited hyaluronic acid binding by melanoma cells, whereas soluble CD44 mutated in the HA binding domain had no effect. When cultivated on hyaluronic acid, melanoma cell proliferation was induced by 30% for both the parental and the control transfected cells. This increase in proliferation was blocked completely in solCD44s-secreting transfectants, whereas solCD44sR41A and solCD44sR150A/R154A-secreting cells again showed hyaluronic acid-induced cell proliferation. These cell lines were subcutaneously injected into MF1 nu/nu mice to compare their growth as tumors in vivo. Compared to tumors derived from parental and control transfected cells, we observed a dramatic reduction of primary tumor growth with solCD44s expressing MM cells. Transfectants expressing solCD44s mutated in the HA binding domain in contrast developed fast-growing primary tumors. These results provide strong evidence that direct solCD44 interactions with hyaluronic acid interfere competitively with processes induced by hyaluronic acid binding to surface CD44. Autocrine, or drug-induced secretion of solCD44 by human melanoma cells may thus exert potent antitumoral effects in vivo. PMID- 11423991 TI - Novel human p53 mutations that are toxic to yeast can enhance transactivation of specific promoters and reactivate tumor p53 mutants. AB - Since highly expressed human p53 can inhibit human and yeast cell growth, we predicted that p53 mutants could be generated with increased growth inhibition of the yeast Saccharomyces cerevisiae and that these would be useful for characterizing p53 functions and tumor p53 mutants. A random mutagenesis screen led to the isolation of mutations in the DNA binding domain that result in p53 being lethal even at moderate expression levels in yeast. Three independent mutants had an alanine change at the evolutionary invariant V122 in the L1 loop. The other toxic mutations affected codons 277 (C277R, C277W) and 279 (G279R). This latter amino acid change was also reported in tumors, while all the other mutations are novel. A recently developed rheostatable GALI promoter system that provides graded increases in expression of p53 was used to examine the transactivation function of the toxic mutations when expression was greatly reduced and cells were viable. At low expression levels the toxic mutants lacked transactivation from a 3xRGC responsive element (RE). Surprisingly some exhibited enhanced transactivation with p21 and bax REs. The V122A mutant was able to re activate transactivation of various p53 tumor mutants and retained growth inhibition when co-expressed with dominant-negative tumor mutations. Upon expression in human Saos-2 cells the V122A p53 mutant caused growth suppression, was capable of transactivation and exhibited higher than wild type activity with the bax promoter in luciferase assays. A non-functional p53 tumor mutant was partially reactivated by V122A for both transactivation and growth suppression. Thus, the screen for toxic p53 mutants in yeast can identify novel p53 variants that may be useful in dissecting p53 regulated cellular responses and in developing p53-based cancer therapies. PMID- 11423992 TI - Photochemical destruction of the Bcl-2 oncoprotein during photodynamic therapy with the phthalocyanine photosensitizer Pc 4. AB - Photodynamic therapy (PDT), utilizing a photosensitizer and visible light, causes localized oxidative damage. With the mitochondrial photosensitizer Pc 4, PDT induces apoptosis, yet its molecular targets are not known. Here, the anti apoptotic protein Bcl-2 is shown to be highly sensitive to PDT, as judged on Western blots by the disappearance of anti-Bcl-2-reactive material from the position of the native 26 kDa protein. The loss of Bcl-2 was PDT dose dependent and was observed for both endogenous and overexpressed Bcl-2 in several cell lines, immediately after PDT, and with chilled cells. It was accompanied by a trace of a 23-kDa cleavage product as well as high-molecular weight products that may result from photochemical crosslinking. PDT-induced Bcl-2 loss occurred in MCF-7 cells that do not express caspase-3 or in the presence of protease inhibitors, but was prevented, along with the induction of apoptosis, by the singlet oxygen scavenger L-histidine. Loss of FLAG-Bcl-2 was observed with both anti-FLAG and anti-Bcl-2 antibodies, indicating loss of native protein rather than simple BCL-2-epitope destruction. Photochemical damage was not observed in Bcl-x(L), Bax, Bad, the voltage-dependent anion channel, or the adenine nucleotide translocator. Therefore, Bcl-2 is one target of PDT with Pc 4, and PDT damage to Bcl-2 contributes to its efficient induction of apoptosis. PMID- 11423993 TI - Lycopene inhibition of cell cycle progression in breast and endometrial cancer cells is associated with reduction in cyclin D levels and retention of p27(Kip1) in the cyclin E-cdk2 complexes. AB - Numerous studies have demonstrated the anticancer activity of the tomato carotenoid, lycopene. However, the molecular mechanism of this action remains unknown. Lycopene inhibition of human breast and endometrial cancer cell growth is associated with inhibition of cell cycle progression at the G(1) phase. In this study we determined the lycopene-mediated changes in the cell cycle machinery. Cells synchronized in the G(1) phase by serum deprivation were treated with lycopene or vehicle and restimulated with 5% serum. Lycopene treatment decreased serum-induced phosphorylation of the retinoblastoma protein and related pocket proteins. This effect was associated with reduced cyclin-dependent kinase (cdk4 and cdk2) activities with no alterations in CDK protein levels. Lycopene caused a decrease in cyclin D1 and D3 levels whereas cyclin E levels did not change. The CDK inhibitor p21(Cip1/Waf1) abundance was reduced while p27(Kip1) levels were unaltered in comparison to control cells. Serum stimulation of control cells resulted in reduction in the p27 content in the cyclin E--cdk2 complex and its accumulation in the cyclin D1--cdk4 complex. This change in distribution was largely prevented by lycopene treatment. These results suggest that lycopene inhibits cell cycle progression via reduction of the cyclin D level and retention of p27 in cyclin E--cdk2, thus leading to inhibition of G(1) CDK activities. PMID- 11423994 TI - Comparative analysis of the ETV6 gene in vertebrate genomes from pufferfish to human. AB - The ETV6 gene encodes an Ets-like transcription factor that is frequently rearranged in leukemias. While some of the functions of ETV6 have been uncovered recently, little is known about the key structural elements involved. Comparative genome analysis may provide novel insights into gene evolution and functions. In this study, we cloned and sequenced the homologue of ETV6 from the compact genome of the pufferfish Fugu rubripes (fETV6). The genomic structure of the fETV6 gene was investigated by sequence analysis of a contig of genomic clones. The fETV6 gene, composed of eight exons, spans about 15 kb and is 16 times smaller than its human counterpart mainly because of the reduced intron size. Three of the seven introns of fETV are unusually large (more than 2 kb), including the 8.2 kb intron 2. The gene codes for a protein of 465 amino acids that is highly related to its human homologue, exhibiting an overall identity of 58% (72% similarity). To investigate the functional and evolutionary aspects of ETV6, we undertook a comparative analysis of this gene from various vertebrates (human, mouse, chicken, zebrafish and Fugu). As expected, the PNT and ETS domains were highly conserved, with on average 81 and 95% peptide sequence identity, respectively. In addition, we found several new highly conserved regions within the central section of the protein that are likely to represent further functional or structural domains, which may be associated with the transcription repression capacity of this protein. We also found conserved putative regulatory elements in the promoter as well as in the large intron 2 of fETV6. The information derived from this comparative analysis will serve as the basis for more precise functional studies of ETV6 gene regulation and function. PMID- 11423995 TI - c-Jun N-terminal kinase activation is required for the inhibition of neovascularization by thrombospondin-1. AB - Thrombospondin-1 (TSP-1) is a potent inhibitor of angiogenesis that acts directly on endothelial cells via the CD36 surface receptor molecule to halt their migration, proliferation, and morphogenesis in vitro and to block neovascularization in vivo. Here we show that inhibitory signals elicited by TSP 1 did not alter the ability of inducers of angiogenesis to activate p42 and p44 mitogen-activated protein kinase (MAPK). Rather, TSP-1 induced a rapid and transient activation of c-Jun N-terminal kinases (JNK). JNK activation by TSP-1 required engagement of CD36, as it was blocked by antagonistic CD36 antibodies and stimulated by short anti-angiogenic peptides derived from TSP-1 that act exclusively via CD36. TSP-1 inhibition of corneal neovascularization induced by bFGF was severely impaired in mice null for JNK-1, pointing to a critical role for this stress-activated kinase in the inhibition of neovascularization by TSP 1. PMID- 11423996 TI - DNA microarray analysis of genes involved in p53 mediated apoptosis: activation of Apaf-1. AB - The transcription regulation activity of p53 controls cellular response to a variety of stress conditions, leading to growth arrest and apoptosis. Despite major progress in the understanding of the global effects of p53 on cellular function the pathways by which p53 activates apoptosis are not well defined. To study genes activated in the p53 induced apoptotic process, we used a mouse myeloid leukemic cell line (LTR6) expressing the temperature-sensitive p53 (val135) that undergoes apoptosis upon shifting the temperature to 32 degrees C. We analysed the gene expression profile at different time points after p53 activation using oligonucleotide microarray capable of detecting approximately 11,000 mRNA species. Cluster analysis of the p53-regulated genes indicate a pattern of early and late induced sets of genes. We show that 91 and 44 genes were substantially up and down regulated, respectively, by p53. Functional classification of these genes reveals that they are involved in many aspects of cell function, in addition to growth arrest and apoptosis. Comparison of p53 regulated gene expression profile in LTR6 cells to that of a human lung cancer cell line (H1299) that undergoes growth arrest but not apoptosis demonstrates that only 15% of the genes are common to both systems. This observation supports the presence of two distinct transcriptional programs in response to p53 signaling, one leading to growth arrest and the other to apoptosis. The proapoptotic genes induced only in LTR6 cells like Apaf-1, Sumo-1 and gelsolin among others may suggest a possible explanation for apoptosis in LTR6 cells. PMID- 11423997 TI - Conflicts of interest. PMID- 11423998 TI - Expression and activation of SHC/MAP kinase pathway in primary acute myeloid leukemia blasts. AB - INTRODUCTION: We report the results of a study investigating signaling proteins in 26 cases of primary acute myelogenous leukemia. We studied the Shc adaptor proteins p52/p46Shc, which can activate the RAS/Mitogen Activated Protein kinase pathway, p66Shc which is uncoupled from RAS/MAP kinases and the MAP kinase family members Extracellular signal Regulated Kinase (ERK) and c-Jun NH2-terminal protein Kinase (JNK) or Stress Activated Protein Kinase (SAPK). MATERIAL AND METHODS: CD34+ and CD34- fractions of four human normal bone marrow and unfractionated bone marrow samples were investigated. Immunoblottings, immunoenzymatic and in vitro assays were performed. RESULTS: Shc protein isoforms were constitutively expressed in all the AML cases examined. Tyrosine phosphorylation of p53/p46Shc isoforms were found in CD34+ but not in the majority of CD34- cases. p66Shc isoform was not tyrosine-phosphorylated in CD34-, and was tyrosine-phosphorylated only in some CD34+ cases. Expression and activation of ERK was constitutively present in the majority of AML patients analysed. JNK/SAPK was expressed but not activated in the AMLs examined. Activation occurred after treatment of the leukemic cells by anisomycin, etoposide, and cytarabine. ERK and JNK/SAPK activation were not detectable in the hematopoietic precursors of human normal bone-marrow. CONCLUSION: These data bear implications for the role of Shc-MAP kinase pathway in normal hemopoiesis and AML leukemogenesis. PMID- 11423999 TI - Factors affecting hematopoietic recovery after autologous peripheral blood progenitor-cell transplantation in aggressive non-Hodgkin's lymphoma: a prospective study of 123 patients. AB - PURPOSE: To analyse prognostic factors influencing hematopoietic recovery in patients with aggressive non-Hodgkin's lymphomas prospectively treated with intensive chemotherapy followed by peripheral blood progenitor-cells transplantation. PATIENTS AND METHODS: Untreated patients with at least two unfavorable factors according to the age-adjusted international prognostic index were included in the LNH 93-3 trial. Patients received three cycles of chemotherapy and PBPC were mobilized using filgrastim. On day 60, a BEAM regimen was initiated followed by PBPC rescue. Among the 123 patients analysed, 60 received G-CSF (5 microg/kg/d) after PBPC transplantation at day 1 and 63 did not. RESULTS: Patients received a mean number of 12.4 x 10(6)/kg (1.86-111.5) CD34+ cells. After transplantation, neutrophil counts exceeded 0.5 x 10(9)/l at a median of 12.4 days (7-41 days) and platelet counts exceeded 50 x 10(9)/l at a median of 15.6 days (9-141 days). Platelets recovery > 50 x 10(9)/l was negatively influenced by BM involvement (20 s 14 days; P = 0.04). The number of CD34+ cells infused (> vs < or = 5 x 10(6)/kg) was correlated with faster platelet recovery (18.7 days vs 13.7 days) (P = 0.007). In 26 patients for whom administration of G-CSF was randomized, time to neutrophil recovery was significantly shorter for patients treated with G-CSF: 10 vs 13 days (P = 0.0005). The incidence of grade 3/4 infection, was similar in both groups. CONCLUSION: In the patient population treated with the same first-line regimen, BM involvement and infusion of fewer CD34+ cells delayed platelet recovery. Administration of G-CSF after PBPC significantly reduced neutropenia. PMID- 11424000 TI - Increased apoptosis in mononucleated cells but not in CD34+ cells in blastic forms of myelodysplastic syndromes. AB - INTRODUCTION: Myelodysplastic syndromes are characterized by peripheral refractory cytopenias together with normo or hyper cellular marrow. Increased apoptosis has been shown to be involved in the process leading to this paradox. MATERIALS AND METHODS: Early apoptosis detection, based on the modification of mitochondrial transmembrane potential (deltapsim), was performed on bone marrow cells from 42 MDS patients (21 RA, four RARS, 10 RAEB, two RAEB-t, three sAML, and two CMML) and seven normal healthy donors. Phosphatidylserine (PS) expression, a late/intermediate marker of the apoptotic cascade, was also quantified. Apoptosis was analysed by flow cytometry both on unsorted mononuclear cells and on progenitor cells after CD34+ magnetic cell sorting. RESULTS: A significant increase of apoptosis of MNC was observed in RA, RARS, RAEB and to a lower extent in RAEB-t and AML samples. In the progenitor compartment, RA and RARS samples presented a high level of apoptosis, whereas a switch to a low level of apoptosis was detected in the blastic forms RAEB, RAEB-t and sAML. Fas (CD95/APO-1), a member of the death domain receptor family, has been reported to be overexpressed on MDS CD34+ marrow cells. A functional assay of Fas cross linking using the CH11 antibody on CD34+ marrow cells was performed on samples of 17 MDS patients; 8/17 were found to be sensitive to Fas-induced apoptosis. However, no correlation was observed with the level of in vivo spontaneous apoptosis. CONCLUSION: This study demonstrates increased apoptosis of MNC in all MDS subgroups as measured by deltapsim collapse. Moreover, while important apoptosis is still observed at the progenitor level in early MDS, blastic forms show a clear reduction of apoptosis. Study of Fas functionality modulates the implication of this receptor in the pathophysiology of the disease. PMID- 11424001 TI - All trans retinoic acid abrogates spontaneous monocytic growth in juvenile chronic myelomonocytic leukaemia. AB - INTRODUCTION: All trans retinoic acid, the active metabolite of vitamin A, exerts profound effects on cell differentiation. On normal myeloid progenitors, retinoids switch the differentiation program of granulo-macrophagic progenitors towards the granulocytic lineage and consequently reduce CFU-M colony formation. Bone marrow and peripheral blood mononuclear cells from children with Juvenile Chronic Myelomonocytic Leukaemia show typical spontaneous monocytic growth. We questioned whether in this disease, retinoids could switch myelomonocytic growth and inhibit the abnormal CFU-M colony proliferation. METHODS: Ten JCML samples were studied in the presence of ATRA in methyl cellulose colony assay, before (CFU-C) or after (pre-CFU) liquid suspension culture. RESULTS: In vitro characteristics of JCML such as spontaneous monocytic growth in the absence of growth factor was noted in all patients. In the presence of leucocyte-conditioned medium, nine samples showed only CFU-M growth and one sample CFU-GM growth. Incubation with ATRA inhibited CFU-M colony formation in nine cases. Enhancement of granulocytic differentiation (CFU-G) was noted in nine cases. ATRA also inhibited CD34+ JCML monocytic growth and GM-CSF hypersensitivity. CONCLUSION: These data suggest that, in JCML progenitors, retinoid pathways are functional and inhibition of immature monocytic progenitors cells may be achieved with retinoids, without impeding granulocytic cell growth. PMID- 11424002 TI - Deregulated expression of prostate apoptosis response gene-4 in less differentiated lymphocytes and inverse expressional patterns of par-4 and bcl-2 in acute lymphocytic leukemia. AB - INTRODUCTION: Prostate apoptosis response gene-4, known as par-4, is a new proapoptotic factor functionally required but not sufficient for apoptosis. Since there is evidence from prostate cancer cells that par-4 is involved in regulation of bcl-2 we assessed expression of par-4 and bcl-2 in different populations of normal and neoplastic lymphocytes. MATERIALS AND METHODS: Expression of par-4 mRNA and protein in different subpopulations of normal and neoplastic lymphocytes was assessed by reverse transcription polymerase chain reaction and Western blot. RESULTS: Par-4 mRNA was not detectable in lymphocytes of healthy volunteers (n = 10), but was present in the majority of samples of chronic lymphocytic leukemia (n = 30), chronic lymphocytic leukemia/prolymphocytic leukemia (n = 6) and acute lymphocytic leukemia (n = 10). Par-4 protein was expressed unanimously in samples of mononuclear cells from healthy volunteers and patients with CLL, but less frequently in immature lymphocytes, including neoplastic cells of CLL/PLL and ALL. The decreased frequency of par-4 expression in immature subpopulations was confirmed by results on lymphocytic cell lines at various stages of maturation. Comparing the expressional patterns of par-4 and bcl-2 there was an inverse relationship of both proteins in ALL and different lymphocytic cell lines, indicating a functional relationship of par-4 and bcl-2. CONCLUSIONS: This study establishes par-4 as a factor expressed in the majority of normal and neoplastic lymphocytic cells, demonstrating a decreased frequency of protein expression in less differentiated lymphocytes and an inverse expressional pattern of par-4 and bcl-2 in lymphocytic cell lines and ALL. PMID- 11424003 TI - Efficient ex vivo expansion of NOD/SCID-repopulating cells with lympho-myeloid potential in hematopoietic grafts of children with solid tumors. AB - INTRODUCTION: The ex vivo expansion of hematopoietic grafts could be an important therapeutic tool for accelerating hematopoietic recovery after administration of high-dose chemotherapy regimens. The fate of the long-term repopulating cells during the ex vivo manipulation of grafts is a critical issue and will ultimately define the clinical applicability of this technology to hematopoietic transplantation. MATERIALS AND METHODS: To study the effects of a clinically applicable ex vivo expansion protocol in the proliferative potential of the most primitive human hematopoietic cells, both LTC-IC and NOD/SCID-RC assays were used to determine LTC-IC and NOD/SCID-RC contents of hematopoietic grafts, both before and after expansion (SCF, IL-3, PEG-MGDF Flt3-L and 5% AB serum), in four children with non-hematological malignancies. RESULTS: The mean percentage of CD34+ cells after expansion was 16%. The numbers of nucleated cells increased 20 fold with a mean three-fold increase in the numbers of CD34+ cells during the expansion period. The CFC content of the samples showed a mean 11-fold increase (range: 5-17) after ex vivo expansion. The primitive hematopoietic stem cell content of the expanded cell fraction evaluated by LTC-IC assays was found to be increased in two patients out of three, with maintenance of the LTC-IC frequency in the third patient. The NOD/SCID-RC potential, evaluated in five experiments from four patients using 109 mice injected 5-6 weeks earlier with human hematopoietic cells, increased from a mean percentage of 36% (range: 7-75%) before expansion, to a mean percentage of 70% (range: 37-100%) after expansion (P < 0.00001). The frequency of NOD/SCID-RC calculated with pooled data from all patients was 1/80,000 at day 0 and 1/40,000 after seven days of culture. The full phenotypic analysis of human hematopoietic cells obtained in NOD/SCID mice injected with expanded cells showed the presence of significant numbers of CD34+, CD19+ and CD15+ cells, suggesting the persistent lympho-myeloid potential of the expanded hematopoietic cells. CONCLUSION: Our results suggest that efficient expansion of NOD/SCID-RC with lympho-myeloid potential can be achieved not only in cord blood or normal marrow as previously reported, but also in hematopoietic grafts obtained from children exposed to high-dose chemotherapy. PMID- 11424004 TI - Risk-oriented postremission strategies in adult acute lymphoblastic leukemia: prospective confirmation of anthracycline activity in standard-risk class and role of hematopoietic stem cell transplants in high-risk groups. AB - INTRODUCTION: Although definite risk classes are well known, risk-adapted modulation of first-line therapy is seldom attempted in adult ALL. So, a prospective validation of the therapeutic efficacy of a protocol (or a component thereof) in specific risk groups is uncommon. MATERIALS AND METHODS: From 1996 1999 a risk-oriented program (08/96) was evaluated in 102/121 unselected patients (median age 35 years, blast count 0-450 x 10(9)/l, 100 B(lin) (lineage), 21 T(lin)) responsive to induction therapy. The standard risk (SR) class was B(lin) CD10+ Ph- with blasts < 10 x 10(9)/l (prior studies: disease-free survival (DFS) rate 52% at five years with dose-intensive anthracycline-containing programs). The SR protocol was therefore anthracycline-rich (early consolidation cycles with total idarubicin 96 mg/m2), and comprised long-term maintenance. High-risk (HR) patients were eligible to the following three options: allogeneic hematopoietic stem cell transplantation (HSCT) from related family donor; short sequence with high-dose cyclophosphamide-cytarabine-methotrexate followed by melphalan/total body irradiation with autologous HSCT; or T(lin) ALL chemotherapy regimen inclusive of high-dose cytarabine and methotrexate. RESULTS: Treatment realization and three-year DFS rates according to risk class, HR subset and postremission treatment intensity were the following. SR group (n = 28): realization rate 93%, DFS 68.5%. HR group (n = 74): realization rate 80%, DFS 39% (P = 0.052 vs SR category). In HR group, three-year DFS rates by disease subtype were the following. B(lin) Ph- (n = 35) 43%; Ph+ (n = 19) 13% at 2.7 years (P = 0.006 vs other HR subtypes); T(lin) (n = 18) 59.5%. And DFS rates by treatment intensity were: allograft (n = 21) 40%; autograft (n = 28) 27%; shift to SR protocol (n = 13) 52% (P = ns vs allograft/autograft); T(lin) program (n = 10) 57%. Matched analyses of treatment protocols and disease subtypes suggested a possible therapeutic role of the autograft regimen in B(lin) Ph- ALL with a blast count < 25 x 10(9)/l, and of T(lin) protocol for T(lin) ALL. Comparisons with retrospective control cohorts were confirmatory of anthracycline activity in SR subclass. CONCLUSION: The intended strategy was applicable to the majority of study patients, confirming the value of anthracyclines in SR class and, preliminarily, the usefulness a T(lin)-specific treatment. Apart from the case of Ph+ ALL, the indications for high-dose procedures with HSCT remains largely undetermined in this study. PMID- 11424005 TI - Retinoic acid dramatically enhances the arsenic trioxide-induced cell cycle arrest and apoptosis in retinoic acid receptor alpha-positive human T-cell lymphotropic virus type-I-transformed cells. AB - INTRODUCTION: Adult T-cell leukemia/lymphoma, caused by the human T-cell lymphotropic virus type I, is an aggressive neoplasm of mature activated T cells that is generally resistant to conventional therapy. While arsenic trioxide (As) inhibits the growth and induces apoptosis in HTLV-I-infected T cells, synergistically, when combined with interferon-alpha, variable effects on growth with all trans retinoic acid treatment have been reported in ATL-derived cell lines and fresh ATL cells. In this study, we investigate the effects of ATRA alone or in combination with As in HTLV-I-transformed cells. MATERIALS AND METHODS: Four HTLV-I-transformed cell lines (HuT-102, MT2, C8166 and C91PL) were treated with different doses of ATRA alone or in combination with As for one to three days. Cell growth was assessed by cell count with 3H-thymidine incorporation. Cell cycle distribution was assessed by propidium iodine-labeled DNA content by flow cytometry. Apoptosis was evaluated by Hoechst nuclear staining and annexin-V binding assays. Expression of retinoid receptors, the viral transactivator Tax, and the proteins bcl-2 and IkappaB-alpha proteins, was analysed by Western blot. RESULTS: Only C8166 cells were sensitive to the ATRA induced growth inhibitory effect while HuT-102, MT2, and C91PL were resistant to ATRA treatment (up to 10(-5) M). The retinoid X receptor alpha and the retinoic acid receptor gamma (RARgamma) proteins were expressed in all four cell lines, while RARalpha protein was only detected in the HuT-102 and C8166 cells. The combination ATRA/As showed a highly synergistic effect on HuT-102 cells, and, to a lesser extent, on C8166 cells and resulted in a dramatic inhibition of cell proliferation and induction of massive apoptosis in HuT-102 cells, associated with caspase activation. While ATRA alone had no effect on Tax and IkappaB-alpha protein levels, ATRA increased the As-induced Tax degradation and the up regulation of IkappaB-alpha protein. In contrast, the expression of bcl-2 protein was not significantly affected by any of the treatments. CONCLUSION: Our data provide a rationale for combined ATRA and As-therapies in ATL patients refractory to conventional therapy and expressing RARalpha in their leukemic cells. PMID- 11424006 TI - Graft-vs-leukemia activity and graft-vs-host disease induced by allogeneic Th1- and Th2-type CD4+ T cells in mice. AB - INTRODUCTION: The transfer of allogeneic lymphocytes contained in a hematopoietic stem cell graft confers an immune-mediated antileukemic effect, termed the graft vs-leukemia (GVL) effect. Graft-vs-host disease (GVHD), the most detrimental complication of allogeneic BMT, largely resides within the same lymphocyte population. Therefore, separation of GVL- and GVH-reactions is a long-standing goal of experimental studies dealing with allogeneic transplantation of hematopoietic stem cells. MATERIALS AND METHODS: The objective of the current study was to assess the potential of Th1- and Th2-type CD4+ T cells in mediating GVHD and GVL effects in a fully allogeneic murine transplant model. BALB/c (H-2d) mice were given a dose of A20 (H-2d, B-cell leukemia) cells two days prior to lethal total body irradiation (TBI) and transplantation of fully mismatched (C57BL/6, H-2b) T-cell depleted (anti-Thy1.2, CD90) bone marrow (TCD-BM) cells. Graded numbers of either unmanipulated, Th1- or Th2-polarized highly enriched CD4+ donor type T cells (10(6) or 10(7)) were administered 2 h posttransplant. Infusion of 10(6) of unmanipulated, Th1-, or Th2-primed CD4+ T cells resulted in moderate GVHD-related mortality (40%, 50%, 10%) and significantly improved long term survival (50%, 45%, 46% surviving the observation period of 120 days) as compared to animals receiving TCD-BM alone (18%). RESULTS: The administration of 10(7) unmanipulated or Th1-type CD4+ T cells given shortly after transplantation led to death of all mice within 50 days due to fatal acute GVHD. In contrast, the adoptive transfer of 10(7) Th2-primed CD4+ T cells resulted in significant improvement of long-term survival (80%) compared to the TCD-BM group. This powerful GVL effect was associated with a substantially lower incidence of lethal acute GVHD (10%) if compared to the results of transplantation of Th1-type CD4+ T cells. CONCLUSION: These results demonstrate that allogeneic Th2-type CD4+ T cells given post BMT can induce GVL effects in a cell-dose-dependent manner without increasing the risk of severe acute GVHD. PMID- 11424007 TI - Definition of an optimal cytotoxic T lymphocyte epitope in the latently expressed Kaposi's sarcoma-associated herpesvirus kaposin protein. AB - Cytotoxic T lymphocytes (CTL) recognize and kill virus-infected cells and contribute to immunologic control of viral replication. For many herpesviruses (e.g., Epstein-Barr and cytomegalovirus), virus-specific CTL responses can be readily detected in infected persons, but CTL responses against Kaposi's sarcoma associated herpesvirus (KSHV) appear to be weak and remain poorly characterized. Using a human leukocyte antigen (HLA) binding motif-based epitope prediction algorithm, we identified 37 HLA-A*0201 binding peptides from 8 KSHV open-reading frames (ORFs). After in vitro stimulation of peripheral blood mononuclear cells from KSHV-infected persons, CTL responses against 1 peptide in the KSHV kaposin protein (ORF K12) were detected in 2 HLA-A*0201-positive subjects. The optimal CTL epitope was identified by HLA restriction analysis and peptide titration assays. These data describe a latent phase viral gene product targeted by CTL that may be relevant for KSHV immunopathogenesis. PMID- 11424008 TI - Antiretroviral therapy effects on genetic and morphologic end points in lymphocytes and sperm of men with human immunodeficiency virus infection. AB - Many human immunodeficiency virus (HIV)-infected persons receive prolonged treatment with DNA-reactive antiretroviral drugs. A prospective study was conducted of 26 HIV-infected men who provided samples before treatment and at multiple times after beginning treatment, to investigate effects of antiretrovirals on lymphocyte and sperm chromosomes and semen quality. Several antiretroviral regimens, all including a nucleoside component, were used. Lymphocyte metaphase analysis and sperm fluorescence in situ hybridization were used for cytogenetic studies. Semen analyses included conventional parameters (volume, concentration, viability, motility, and morphology). No significant effects on cytogenetic parameters, semen volume, or sperm concentration were detected. However, there were significant improvements in sperm motility for men with study entry CD4 cell counts >200 cells/mm(3), sperm morphology for men with entry CD4 cell counts < or =200 cells/mm(3), and the percentage of viable sperm in both groups. These findings suggest that nucleoside-containing antiretrovirals administered via recommended protocols do not induce chromosomal changes in lymphocytes or sperm but may produce improvements in semen quality. PMID- 11424009 TI - Protection from secondary human immunodeficiency virus type 1 infection in chimpanzees suggests the importance of antigenic boosting and a possible role for cytotoxic T cells. AB - Recent evidence suggests a much higher prevalence of human immunodeficiency virus type 1 (HIV-1) recombinants than previously anticipated. These recombinants arise from secondary HIV infections in individuals already infected with the virus. It remains unclear why some individuals acquire secondary HIV-1 infections and others do not. To address this question, a study was undertaken of a small cohort of chimpanzees with well-defined HIV-1 infection. After exposure to an infectious dose of heterologous primary isolate, 4 of 8 HIV-1 seropositive chimpanzees resisted secondary infection, whereas 2 naive controls became readily infected. Only animals who were immunologically boosted were protected. Protection from heterologous secondary exposure appeared to be related to the repertoire of the cytolytic CD8(+) T cell responses to HIV-1. Data suggested that immunologic boosting by HIV-1 antigens or exposure to subinfectious doses of virus may be important events in sustaining sufficient immunity to prevent secondary infections from occurring. PMID- 11424010 TI - Capsulation loci of non-serotype b encapsulated Haemophilus influenzae. AB - Invasive infections caused by non-type b encapsulated Haemophilus influenzae have increased recently. Because capsule is a major virulence factor, capsulation of 62 recently isolated non-serotype b encapsulated strains was examined. Repeated serotyping confirmed only 69.0% of isolates. The combination of slide agglutination and cap genotyping confirmed 78.9% of type a, 100% of type e, and 86.4% of type f strains. Seven nonencapsulated strains may have lost capsulation through homologous recombination. Three strains that could not be serotyped or genotyped failed to hybridize with any cap probe and are probably nontypeable H. influenzae. Of isolates that retained an intact cap locus, 62.2% had evidence of cap amplification. The epidemiology of non-type b encapsulated H. influenzae infections is complicated by the poor specificity of available serologic reagents and by spontaneous capsule loss. Recently isolated invasive non-type b encapsulated H. influenzae frequently have cap amplification, which may contribute to their virulence. PMID- 11424011 TI - The importance of a beta-glucan receptor in the nonopsonic entry of nontypeable Haemophilus influenzae into human monocytic and epithelial cells. AB - Previous reports showed that nontypeable Haemophilus influenzae (NTHi) reside in macrophage-like cells in human adenoid tissue. This study investigated the ability of nonopsonized NTHi and encapsulated H. influenzae type b (Hib) to enter human monocytic and epithelial cells. The number of intracellular bacteria was determined by a viability assay and flow cytometry. To characterize the mechanisms responsible for the internalization of NTHi, different inhibitors of surface molecules, receptor turnover, and the cytoskeleton were used. Hib were found in monocytic cells at very low numbers (<100 bacteria/2x105 cells). In contrast, a great variation in intracellular numbers was detected between the different NTHi isolates (range, 0.0007%-0.28% of the inoculum for monocytes and 0.053%-3.5% for epithelial cells). NTHi entered human monocytic and epithelial cells via a receptor-mediated endocytosis involving mainly a beta-glucan receptor that could be blocked by laminarin. PMID- 11424012 TI - Strain-specific iron-dependent virulence in Escherichia coli. AB - For reasons unknown, certain Escherichia coli strains become highly virulent when injected with hemoglobin or other soluble iron sources. Two clinical isolates (virulent and nonvirulent) showed equivalent hemoglobin-mediated growth acceleration in vitro. However, when injected intraperitoneally into mice without hemoglobin, the virulent strain was cleared more slowly (t(1/2), >4 h vs. <30 min). The virulent E. coli strain had a polysialic acid-containing capsule, whereas the nonvirulent strain did not. Virulent E. coli grown at 20 degrees C (which blocks polysialylation) were cleared as rapidly as nonvirulent organisms. In another virulent E. coli strain having abundant outer membrane polysialic acid, targeted deletion of the polysialyltransferase accelerated host clearance and blocked iron-dependent virulence. The iron-dependent virulence of certain E. coli strains may represent the combined effect of slow in vivo clearance associated, in this case, with outer membrane polysialylation coupled with accelerated growth permitted by iron compounds. PMID- 11424013 TI - Enhanced susceptibility to superantigen-associated streptococcal sepsis in human leukocyte antigen-DQ transgenic mice. AB - Bacterial superantigens are believed to cause septic shock, although, because of the lack of superantigen-sensitive infection models, proof that superantigenicity underlies shock pathogenesis is lacking. This work demonstrates a clear superantigen effect in septic shock resulting from bacterial infection. Transgenic expression of human leukocyte antigen (HLA)-DQ, but not HLA-DR, specifically augments lymphocyte responses to streptococcal pyrogenic exotoxin A (SPEA). HLA-DQ transgenic mice had increased mortality after administration of SPEA or infection with Streptococcus pyogenes. Immune activation during infection was HLA-DQ transgene-dependent and was manifested by Vbeta-specific T cell repertoire changes and widespread lymphoblastic tissue infiltration. Unlike earlier models, which used toxin-induced shock, these T cell superantigen responses and lymphoblastoid changes were observed during invasive streptococcal sepsis. Lymphoid activation was undetectable in HLA-DQ mice infected with an isogenic SPEA(-) strain, which proves that a single superantigen can play a role in sepsis pathogenesis. PMID- 11424014 TI - Differences in synovial fluid levels of matrix metalloproteinases suggest separate mechanisms of pathogenesis in Lyme arthritis before and after antibiotic treatment. AB - The cause of persistent arthritis in patients with Lyme disease who have received standard antibiotic therapy remains an area of debate. In this study, synovial fluid levels of matrix metalloproteinases (MMPs) were compared in persons with untreated and antibiotic-resistant Lyme arthritis. Levels of MMP-1 and MMP-3, as determined by ELISA, were higher in untreated patients (P=.0064 and P=.002, respectively), whereas levels of MMP-8 and MMP-9 were higher in antibiotic resistant patients (P=.0002 and P=.0014, respectively). In vitro studies of chondrocyte cultures infected with Borrelia burgdorferi revealed induction of MMP 1 and MMP-3 but not of MMP-8 or MMP-9. Neither Staphylococcus aureus nor lipopolysaccharide stimulated MMP-1 or MMP-3 release from these cells. The mechanism of recognition of B. burgdorferi may be through CD14 and toll-like receptor-2, which were up-regulated in the presence of B. burgdorferi. These findings suggest different stimuli for MMP induction in untreated and antibiotic resistant Lyme arthritis. PMID- 11424015 TI - Heparan sulfate-like glycosaminoglycan is a cellular receptor for Chlamydia pneumoniae. AB - Chlamydia pneumoniae is an important human intracellular pathogen; however, the pathogenesis of C. pneumoniae infection is poorly understood, and the bacterial adherence mechanism to host cells is unknown. This study examined the role of glycosaminoglycans (GAGs) in the adhesion of C. pneumoniae to eukaryotic cells. Heparin and heparan sulfate were found to inhibit the attachment of C. pneumoniae to human epithelial cells. Reduction in infectivity resulted from the binding of heparin to the organism. Enzymatic removal of heparan sulfate moieties from the host cell surface led to a marked decrease in C. pneumoniae infectivity. Mutant CHO cell lines that were defective in heparan sulfate biosynthesis were less susceptible to C. pneumoniae infection than was the wild-type cell line. However, preincubation of the GAG-deficient CHO cells with exogenous heparin greatly increased infectivity. PMID- 11424016 TI - Influenza revaccination of elderly travelers: antibody response to single influenza vaccination and revaccination at 12 weeks. AB - The antibody response to a single influenza vaccination and the effect of influenza revaccination was assessed in healthy elderly persons. Travelers > or =65 years old who had received influenza vaccine before travel were enrolled in the study and were offered a second vaccination after 12 weeks. Geographic and age-matched control subjects received a single vaccination. A second influenza vaccination was not associated with increased adverse effects. There was no significant difference between log(10) hemagglutinin-inhibiting (HI) antibody titers or an HI antibody titer > or =1:40 (considered to be protective) in 28 control subjects and 28 revaccinated travelers for any antigen. Probable protection for influenza A antigens remained high 24 weeks after a single immunization and revaccination (A/Sydney/05/97 [H3N2], 92% and 96%, and A/Beijing/262/95 [H1N1], 80% and 96%, respectively). Response to B/Harbin was less throughout the study. A/Sydney antibody titer was lower with more times vaccinated in the previous 5 years. Therefore, a second vaccine did not enhance the immune response. PMID- 11424017 TI - Replication of human cytomegalovirus in severe combined immunodeficient mice implanted with human retinal tissue. AB - Because human cytomegalovirus (HCMV) infection and replication are limited to human cells, few animal models can be used to specifically examine the biology of HCMV in vivo. In these studies, fetal human retinal tissue was implanted into the anterior chamber of the severe combined immunodeficient (SCID) mouse eye and subsequently was inoculated with HCMV. Viral replication, localized to glial cells in the xenografts, was first detected 7 days after infection. Thereafter, HCMV replication increased to peak levels through days 21-28 and then gradually decreased to undetectable levels by 8 weeks after infection. The clinical isolate Toledo replicated to higher titers than did strain AD169 or Towne. A comparison of implant age indicated that older tissue could support higher levels of HCMV replication than could younger implants. SCID mice implanted with human retinal tissue provide an excellent model for evaluation of HCMV infection of an ocular structure in vivo. PMID- 11424018 TI - Application of a topical immune response modifier, resiquimod gel, to modify the recurrence rate of recurrent genital herpes: a pilot study. AB - Resiquimod (R-848), a topically active immune response modifier, induced production of interferon-alpha and interleukin-12 in cultured blood mononuclear cells and decreased genital herpes recurrences in an animal model. In this study, 52 patients with frequently recurrent genital herpes applied topical resiquimod gel 0.01% (twice or thrice weekly) or 0.05% (once or twice weekly) or vehicle gel to herpes lesions for 3 weeks. During the 6-month observation period after treatment, median days to first recurrence in the pooled resiquimod group was 169 days, compared with 57 days for the vehicle group (P=.0058). In all, 32% of resiquimod-treated patients completed the observation period without a recurrence, compared with 6% of vehicle-treated patients (P=.039). Resiquimod 0.05% twice weekly produced dose-limiting inflammation at the lesion sites, but the other regimens were well tolerated. Application of resiquimod to genital herpes lesions appeared to reduce the frequency of recurrences. PMID- 11424019 TI - Long-term protease inhibitor-containing therapy results in limited improvement in T cell function but not restoration of interleukin-12 production in pediatric patients with AIDS. AB - This study investigated whether immune restoration occurred in 26 human immunodeficiency virus (HIV) type 1-infected children treated first with indinavir for 16 weeks and then with combination antiretroviral therapy for >2 years. Compared with baseline, a significant, although modest, decrease in virus loads (maximum median, -0.86 log(10)) and increase in the number of CD4(+) lymphocytes, especially naive cells, were observed at several time points after 2 years. A maximum of 7% of treated children achieved undetectable viremia. There was a marked increase in the proliferative response and skin reactivity to recall antigens. However, responses to an HIV antigen remained depressed, and the production of interleukin-12 remained unchanged and abnormally low. The magnitude of virus suppression did not correlate with these measures of functional immune reconstitution. These findings suggest that long-term nonsuppressive antiretroviral therapy can induce limited improvement in immune function in pediatric AIDS patients and that the effect of suppressive treatments should be investigated. PMID- 11424020 TI - A monkey model for enterohemorrhagic Escherichia coli infection. AB - Adult Macaca radiata (n=22) were infected intragastrically with 10(12) Escherichia coli O157:H7 strain 84-01, which produces Shiga toxins 1 and 2. Clinical symptoms and bacterial excretion were documented in each monkey for a specified time period before they were killed. At necropsy, samples were obtained for culture and histologic and ultrastructural examination. Seventeen monkeys had diarrhea: E. coli O157 was isolated from postinfection stool samples from all monkeys and from autopsy cultures for 14 of 22 monkeys. Histologic examination showed attaching-effacing lesions, which appeared at 12 h and persisted for 7 days, in 12 monkeys. Widening of the intercellular spaces, degeneration and vacuolization of the epithelial cells, epithelial tufting, extrusion of epithelial cells, and neutrophilic infiltration were characteristic features seen in 20 of the 22 infected monkeys but not in 4 control monkeys. This monkey model closely parallels the early stages of the disease produced by E. coli O157:H7 and would be useful in the further study of pathogenic mechanisms and prevention methods in enterohemorrhagic E. coli infections. PMID- 11424021 TI - Enterobacter species in a pediatric hospital: horizontal transfer or selection in individual patients? AB - Enterobacter species were studied longitudinally in a children's hospital. In total, 287 Enterobacter isolates were obtained from 171 children in 15 different wards (from March 1995 through April 1997). Strains were typed by random amplified polymorphic DNA and pulsed-field gel electrophoresis, which were concordant in outcome. In total, 97 DNA types and 199 colonization events were identified. A predominant clone was isolated 111 times from 62 children; another clone was isolated 19 times from 10 patients. These clones caused 36% of all colonizations. In 34% of the children, Enterobacter clones were found in 2-4 patients. The remaining colonizations were due to unique Enterobacter isolates. A large proportion of the Enterobacter strains was acquired through cross transmission. This finding contrasts with the prevailing opinion that resistant Enterobacter strains are selected primarily from the patient's own gut flora. PMID- 11424022 TI - Bacterial infection promotes colon tumorigenesis in Apc(Min/+) mice. AB - The Min mouse, which has a germ line mutation in 1 allele of the Apc tumor suppressor gene, is a model for the early steps in human colorectal cancer. Helicobacter pylori infection, a known risk factor for gastric cancer in humans, causes chronic inflammation and increased epithelial cell proliferation in the stomach. Infection with the bacterium Citrobacter rodentium is known to increase epithelial cell proliferation and to promote chemically initiated tumors in the colon of mice. Min mice infected with C. rodentium at 1 month of age were found to have a 4-fold increase in the number of colonic adenomas at 6 months of age, compared with uninfected Min mice. Most of the colonic adenomas in the infected Min mice were in the distal colon, where C. rodentium-induced hyperplasia occurs. These data demonstrate that bacterial infection promotes colon tumor formation in genetically susceptible mice. PMID- 11424023 TI - Interleukin-12 receptor beta1 deficiency in a patient with abdominal tuberculosis. AB - Two siblings with interleukin-12 receptor beta1 (IL-12Rbeta1) deficiency but different clinical phenotypes were studied. Both are homozygous for an IL12RB1 missense mutation that prevents receptor expression and abolishes cellular responses to IL-12. Transfection of the patients' T cells with wild-type IL12RB1 restored IL-12Rbeta1 expression and function. One patient had the expected phenotype of disseminated bacille Calmette-Guerin (BCG) infection in early childhood, whereas the other did not develop BCG infection, despite 3 inoculations with live BCG. Abdominal tuberculosis was diagnosed in this second patient at age 18 years. To date, neither of them has had clinical disease caused by environmental mycobacteria. These observations show unexpected interfamilial and intrafamilial heterogeneity of the clinical phenotype associated with IL 12Rbeta1 deficiency. The patients may be resistant to BCG but remain vulnerable to Mycobacterium tuberculosis. A diagnosis of IL-12Rbeta1 deficiency should therefore be considered in selected patients with severe tuberculosis, despite their resistance to BCG and a lack of atypical mycobacteriosis. PMID- 11424024 TI - Association of severe malaria with a specific Plasmodium falciparum genotype in French Guiana. AB - Why severe Plasmodium falciparum malaria occurs in only a small percentage of patients is unclear. The possibility that specific parasite characteristics contribute to severity has been investigated in French Guiana, a hypoendemic area, where parasite diversity is low and all patients with severe cases are referred to a single intensive care unit. Parasite genotyping in geographically and temporally matched patients with mild and severe disease showed that the association of a specific msp-1 allele (B-K1) with a specific var gene (var-D) was overrepresented among patients with severe versus mild disease (47% vs. 3%, respectively; P<.001). Moreover, this genotype combination was consistently observed in the most severe clinical cases. Reverse-transcription polymerase chain reaction demonstrated programmed expression of var-D in vivo, which is consistent with its potential implication in severe disease. These results provide field evidence of an association of severe malaria with specific genetic characteristics of parasites and open the way for intervention strategies targeting key virulence factors of parasites. PMID- 11424025 TI - Limitations of pulsed-field gel electrophoresis for the routine surveillance of Campylobacter infections. PMID- 11424027 TI - [Focal cerebral ischemic or hemorrhagic lesions in the term newborn. Review of the last decade]. AB - INTRODUCTION: Although focal cerebral ischemic or hemorrhagic lesions are infrequent in the term newborn, they must be considered when neurologic symptoms appear, especially when seizures are present. OBJECTIVE: Possible risk factors to suffer from these pathologies have been studied, as well as their evolution, to try to give a prognosis. PATIENTS AND METHODS: The term newborns with focal ischemic or hemorrhagic cerebral lesions who presented symptoms in the neonatal period have been studied for 10 years (January 1990-March 2000) in our Children s Hospital Miguel Servet of Zaragoza (Spain). The newborns have been studied in our hospital, and their evolution followed in the Neuropediatric Consulting Room. From each case data about familiar history, pregnancy, labor, clinical manifestations, physical examination, complementary studies, diagnosis and neurodevelopment evolution have been collected. RESULTS. From the nine cases found, four were infarctions of the left medial cerebral artery, and five were hemorrhages. Except in two cases in which an important birth trauma was present, any other antecedent that could be the cause was found. All except one manifested as seizures. Neuroimaging studies visualized the stroke in all of them. A slight motor deficit remains in seven children, and it is severe in one. CONCLUSIONS: Cerebrovascular strokes are infrequent in term newborns. Most of the times their etiology is not found. These accidents usually manifestate as focal seizures in the immediate neonatal period, and neuroimaging studies (ECO-TC) are essential for the diagnosis. The long term evolution is favorable in most of the cases, although focal motor deficit remains present frequently. PMID- 11424028 TI - [Risk factors involved in symmetrical distal diabetic neuropathy]. AB - INTRODUCTION: Diabetic neuropathy is a common neurological complication in patients with diabetes mellitus which affects their quality of life. The risk factors involved in this are not completely clear. OBJECTIVE: To determine the frequency of symmetrical distal diabetic neuropathy and the risk factors associated with its occurrence in the group studied. PATIENTS AND METHODS: A transversal study was made in the Hospital Clinico Quirurgico Hermanos Ameijeiras of Ciudad de la Habana (Cuba), in 200 patients diagnosed as having type 1 or type 2 diabetes mellitus, between January 1997 and December 1999. A clinical neurological examination and neurophysiological investigations were done on all patients to make a diagnosis of diabetic neuropathy. Subsequently, we compared those with diabetic neuropathy and those without in relation to the risk factors. Statistical analysis included the chi squared test and estimation of the relative risk. RESULTS. The frequency of symmetrical distal diabetic neuropathy was 30.0%. Related risk factors were: smoking (p= 0.00, RR= 2.59), raised serum cholesterol (p= 0.00, RR= 4.12), poor metabolic control of diabetes mellitus (p= 0.00, RR= 4.13), associated disorders (p= 0.00, RR= 5.31) including arterial hypertension (p= 0.03, RR= 1.90) and duration of diabetes mellitus (p= 0.00, RR= 2.01). CONCLUSIONS: In our study symmetrical distal diabetic neuropathy was a common complication of diabetes mellitus associated with various risk factors. PMID- 11424029 TI - [Joubert's syndrome: report of 12 cases]. AB - INTRODUCTION: Joubert syndrome is a rare autosomal recessive disorder whose main clinical signs are hypotonia, ataxia, mental retardation, abnormal eye movements and a respiratory pattern of alternating tachypnea-apnea during first months of life. The most characteristic imaging features are elongation and thinning of the pontomesencephalic junction with deepening of the interpeduncular fosse, thickening of the superior cerebellar peduncles, hypoplasia of the vermis and incomplete fusion of the halves of the vermis, creating a sagittal vermic cleft. The first three findings are components of the molar tooth sign . OBJECTIVES: Our aim was to review the clinical features and the neuroradiological findings in 12 children with clinical diagnosis of Joubert syndrome, along with the attempt to correlate clinical and radiological findings. PATIENTS AND METHODS: We performed a retrospective study, and cerebral magnetic resonance imaging was achieved in all cases. RESULTS: All the children have mental retardation, hypotonia, ataxia and oculomotor abnormalities. Other clinical findings are respiratory rhythm abnormalities, abnormal retinal pigmentation, mouth-tongye-facial dyskinesias, ptosis, polydactyly, scoliosis, congenital heart defects, polycystic kidneys and seizures. All patients have agenesis of the vermis and the molar tooth sign is present in nine patients. Five children have other associated cerebral malformations. CONCLUSIONS: In the absence of a biochemical or genetic marker for the Joubert syndrome, we need to have a group of patients with homogeneous clinical and neuroradiological characteristics, in order to avoid an overlap with other syndromes. According to our experience and the review of the literature, we believe that the following should be considered as major diagnostic criteria for Joubert syndrome: hypotonia, ataxia, mental retardation, oculomotor apraxia and the molar tooth sign . Supporting clinical features are: abnormal respiratory pattern, retinal pigmentation, renal abnormalities and facial dysmorphism. PMID- 11424030 TI - [Neuroprotection in acute ischemic stroke. Practicability of guidelines for treatment]. AB - INTRODUCTION: Fibrinolytic agents are effective in the treatment of acute ischemic stroke. However, logistic and clinical factors limit their use. Neuroprotective drugs pose less risks and can be used even before performance of computed tomography of the brain as they are not detrimental in hemorrhagic stroke. These aspects, in theory, will allow the use of neuroprotective drugs in larger number of patients. OBJECTIVE: To evaluate the feasibility of a neuroprotection protocol and the potential usefulness of citicoline in acute ischemic stroke. PATIENTS AND METHODS: Thirty seven patients admitted with a clinical diagnosis of acute ischemic stroke (later confirmed with computed tomography) received, within 12 hours of onset of symptoms, citicoline 500 mg intravenously in a single bolus daily for 7 days. Neurological outcome in this group was compared with a group of 37 patients admitted during the 6 month period before the initiation of the trial and not treated with citicoline. Groups were matched by National Institute of Health Stroke Scale (NIHSS) on admission. RESULTS: Patients treated with citicoline (aged 69+/-14 years) improved on their NIHSS from admission (5.7+/-4.2) to discharge (4.7+/-4.5), p= 0.015. The control group (aged 60+/-17 years) did not change between admission (5.7+/-4.3) and discharge (5.2+/-3.5), ns. Patients treated within 6 hours of admission (n= 12) had more substantial improvement, from 5.4+/-2.3 on admission to 3.9+/-2.9 at discharge, p= 0.008. There were no differences in vascular risk factor profile between the groups. Citicoline was well tolerated in every subject. CONCLUSIONS: A protocol of acute stroke management using neuroprotective agents presents clear logistic advantages allowing the inclusion of larger number of patients. Citicoline appears as a safe and potentially effective option. PMID- 11424031 TI - [Chronic and subacute subdural haematoma. An epidemiological study in a captive population]. AB - INTRODUCTION: Although chronic and subacute subdural haematomas (CSSH) are amongst the commonest neurosurgical conditions, there are few studies on their incidence in the general population. OBJECTIVES: To determine the overall annual rate, the specific rates according to age and sex based on the Official Argentinian National Census of 1991 (OANC 91) for CSSH. PATIENTS AND METHODS: The Hospital Privado de Comunidad de Mar del Plata attends a captive population of 89,500 persons from the Instituto Nacional de Servicios Sociales de Jubilados y Pensionados (INSSJP) and the Prepaid Medical Schemes (PMP) of our institution. We studied the patients of INSSJP and PMP who had CSSH between 1992 and 1996. We determined the annual overall rate and the specific rates according to age and sex, and fitted to the OANC 91. RESULTS: 1. Annual overall rate: 14.1 CSSH/100,000 persons/year. 2. Specific rate for women: 11.6 CSSH/100,000 persons/year. 3. Specific rate for men: 18.1 CSSH/100,000 persons/year. 4. Specific rate 71-80 years old: 18.8 CSSH/100,000 persons/year. 5. Rate fitted to OANC9: 3.1 CSSH/100,000 persons/year. CONCLUSIONS: Our overall rate is higher, and the specific rate for the age group 71-80 years is intermediate, with regard to the rates found in other studies. Neuroepidemiological investigation should be stimulated so that more clinical studies are made regarding the results and costs based on the population. PMID- 11424032 TI - [Endogenous nerve growth factor in patients with Alzheimer s disease]. AB - INTRODUCTION: Nerve growth factor (NGF) is the best characterized neurotrophic polypeptide. Initially it was postulated that alterations in the expression of NGF within its production sites may be responsible for the consistent atrophy and loss of cholinergic basal forebrain neurons in dementing illness such as Alzheimer s Disease (AD). OBJECTIVE: We analyze the NGF concentration in serum from control and patients with AD in order to elucidate something alteration in this fluid related with AD neuropathology. PATIENTS AND METHODS: We applied a twosite enzyme immunoassay to determine NGF levels in sera of patients with AD. RESULTS: Sera from Alzheimer s patients (36+/-7 pg/ml) showed slight but non significant reduction in NGF levels when compared with age related controls (66+/ 18 pg/ml). On the another hand, the NGF concentrations in AD patients and control subjects to the sex were following: female AD patients showed a mean of 33.27+/ 10.43 pg/ml vs 57.83+/-22 pg/ml founded in female controls, while the mean value for male AD patients was 40.87+/-12.29 pg/ml vs 37.47+/-12.28 pg/ml for the male controls. In all the cases studied, no significant differences were observed according to Student t-Test. CONCLUSIONS: Even when no significant differences were observed between controls and AD patients, the results show a tendency NGF concentration decrease in this illness. Certainly, NGF is produced not only in the forebrain but throughout the body, for this reason, more studies, including the analysis of cerebrospinal fluid would be useful to define the real relationship between NGF concentration changes in serum and AD-related changes in endogenous NGF concentrations, taking into account increasing levels by exogenous NGF administration could still be useful in maintaining the cholinergic neurons. PMID- 11424033 TI - [Morbi-mortality of medical and surgical treatments in the intracranial bleeding like complication of therapy with drugs that modifies normal coagulation]. AB - INTRODUCTION: Antiaggregant and anticoagulant therapy has possible sever secondary effects, being the most serious intracranial bleeding. OBJECTIVES: Compare morbi-mortality between surgical and medical treatments in anticoagulated and/or antiaggregated patients with any kind of intracranial bleeding. Also risk factors and main indications of those treatments are studied. PATIENTS AND METHODS: Descriptive and retrospective study including 105 patients admitted in our Critical Care Unit, with intracranial bleeding, under anticoagulant or antiaggregant treatment. We study the type of bleeding (subarachnoid, epidural, subdural and intraparenchimal bleeding) and the relation between morbi-mortality and treatment applicated using relative risk like analytic tool. RESULTS: Major bleeding risk appears after the six first months from the beginning of the treatment. Thirty-four patients died with a relative risk of 1.55 in the surgical patients from the medical treatment patients. Relative risk between anticoagulated patients and antiaggregated ones was 1.16. Serious consequences happened on eight of the 49 patients under surgical treatment, and on 13 of the 52 patients under medical treatment. CONCLUSIONS: Surgical treatment has more morbi-mortality. Oral anticoagulation has only a little more relative risk than treatment with antiaggregants. Both groups had serious consequences. PMID- 11424034 TI - [Intradural spinal arachnoid cyst associated with Noonan's syndrome]. AB - INTRODUCTION: Intradural spinal arachnoid cysts are collections of liquid similar to CSF, caused by a disorder of the arachnoid trabeculae. Noonan s syndrome is a condition involving multiple malformations, with autosomal dominant inheritance, variable penetration and expression and a phenotype similar to that of Turner s syndrome. We describe the case of a girl with Noonan s syndrome who presented with progressive scoliosis and signs and symptoms of a disorder of the spine related to an intradural spinal arachnoid cyst. CLINICAL CASE: A four year old girl with no significant previous clinical history was seen for slow growth (in size and weight), progressive scoliosis and urinary sphincter dysfunction. On physical examination there were features of polymalformation compatible with Noonan s syndrome and thoracic scoliosis. In view of this a spinal MR study was done. This showed an intradural arachnoid cyst from segment T4 to T11, with displacement and spinal narrowing which required a cyst-peritoneal shunt. CONCLUSIONS: Intradural spinal arachnoid cysts are collections of liquid produced due to arachnoid trabeculae. Noonan s syndrome is a condition involving multiple malformations with a phenotype which is very similar to that of Turner s syndrome. It is linked to alterations of tissue elastin which favor the appearance of arachnoid cysts. Arachnoid cysts are an uncommon cause of spinal compression and/or progressive scoliosis. Although this association has not been described previously, in the case of a patient with Noonan s syndrome, with clinical features of a spinal disorder and/or progressive scoliosis, the presence of a spinal arachnoid cyst should be considered. PMID- 11424035 TI - [Permanent visual loss resulting from bilateral occipital infarction after shunt malfunction]. AB - INTRODUCTION: The commonest cause of malfunction of shunts for cerebrospinal fluid is obstruction of the catheter, which may be of subacute or sudden onset. When onset is sudden, the complication is serious as this implies risk of death or permanent damage to vision. CLINICAL CASE: We describe the case of a 13 year old girl who, 8 days after insertion of a ventriculo-peritoneal shunt valve, complained of the sudden onset of headache, reduced level of consciousness, pupil changes and permanent loss of vision secondary to a bilateral occipital infarct. CONCLUSIONS: The visual pathways and oculomotor nerves are particularly sensitive to changes in intracranial pressure and displacement of intracranial structures. Therefore, neuro-ophthalmic signs are more sensitive than computerized tomography (CT) in detection of mechanical malfunction of the valve. Although CT shows structural anomalies, it cannot assess their function. The intracranial pressure may be high enough to cause damage to the visual pathways, although no enlargement of the ventricles can be seen on CT. A normal CT, or one similar to previous scans, does not rule out malfunction of the shunt. If clinical signs of raised intracranial pressure are observed, the possibility of mechanical malfunction of the valve should be considered and this should be explored as a neurosurgical emergency in view of the risk of death or permanent loss of vision. PMID- 11424036 TI - [Polyneuropathy in critically ill patients: a seldom recognized cause of dependence on mechanical ventilators]. AB - INTRODUCTION: Neuromuscular complications, in a critical care unit, are a cause of morbidity in children and prolonged dependence on a mechanical ventilator. Polyneuropathy of the critical patient is such a complication and is seen in patients on mechanical respiratory assistance. OBJECTIVE: To discuss the neurological and electrophysiological clinical findings of polyneuropathy of the critical patient. CLINICAL CASE: We evaluated four patients who initially required mechanical respiratory assistance, three for lung disorders and one for acute encephalopathy, who developed prolonged dependence on mechanical ventilators in the year 1999. In all these patients electromyography showed primary axon nerve disorders with secondary demyelination of all four limbs and phrenic nerve involvement. CONCLUSIONS: Clinical suspicion and use of suitable electrophysiological techniques permits identification of this condition in the severely ill paediatric patient. Better recognition of the condition and investigation of the etiological factors would help to develop suitable measures for prevention and treatment. PMID- 11424037 TI - [Hypoxic encephalopathy and cortical laminar necrosis]. AB - INTRODUCTION: Cortical laminar necrosis is characterized by destruction of the cerebral cortex, mainly of the third layer, in situations of reduced energy supply to the brain. The cerebral lesions caused are known through studies made at autopsies, but there are few descriptions in the literature of the neuroimaging changes. We report the case of a patient who suffered hypoxic encephalopathy secondary to prolonged status epilepticus, and in whom cerebral MR showed changes compatible with cortical laminar necrosis. CLINICAL CASE: A 16 year old girl who had been epileptic since infancy presented with a state of generalized tonic-clonic convulsions followed by coma. Three weeks later she was mute, had a blink reflect to threats and followed visual stimuli with eye movements but no voluntary motor or verbal response. She also showed generalized hypertonia and fine tremor of her arms, which she moved spontaneously with no asymmetry. After two months her clinical condition became stable. Cerebral MR at this time showed diffuse hypersignal of the cortex and basal ganglia in T2 and FLAIR sequences and hyposignal of the subcortical white matter associated with a marked hypersignal delimiting the grooves of convexity in T1 sequences. CONCLUSIONS: Situations of prolonged hypoxia, such as in status epilepticus, lead to necrosis of layers of the cerebral cortex. Clinically this is seen as the appearance of hypoxic encephalopathy and radiologically as characteristic alterations of neuroimaging known as cortical laminar necrosis. PMID- 11424038 TI - [Apoptosis in neurodegenerative diseases: facts and controversies]. AB - OBJECTIVES: In this article, the authors analyzed critically the morphological and biochemical evidences of cell death by apoptosis from postmortem studies in Alzheimer s, Parkinson s, Huntington s and Wilson s diseases. DEVELOPMENT: During the last few years, apoptosis has been postulated as a type of neuronal death responsible for the neurodegenerative process in those heterogeneous, chronic and progressive neurological disorders, which are characterized by a selective and a symmetric loss of neurons in motor, sensory or cognitive systems. With regard to neuronal death mechanism and the contribution of the mutated or metabolic altered proteins such as betaA, P-tau, alpha-synuclein, Parkin, Huntingtin, ATP78B, proteins in the pathogenesis of those disorders are still unknown. CONCLUSIONS: We consider that the morphological (e.g. DNA fragmentation without showing classical apoptotic morphology) and biochemical evidences are still insufficient and contradictory to formally indict apoptosis as the mechanism of neuronal cell death in those neurological disorders. The establishment of the molecular mechanisms leading neurons to cell death (by apoptosis?) could provide significant information for the design of therapeutic strategies to retard or prevent the development of such neurodegenerative diseases in affected individuals. PMID- 11424039 TI - [Therapeutic strategy in severe encephalopathies]. AB - INTRODUCTION: The severe epileptic syndromes of infancy are dependent on age, symptomatic or cryptogenic aetiology and are drug-resistant so that overall prognosis control of epilepsy and adequate cognition is poor so that some authors consider it to be catastrophic. All may be identified and differentiated on electroclinical criteria. DEVELOPMENT: We study the neonatal myoclonic encephalopathies in the two clinical forms described by Aicardi and Ohtahara, West s syndrome (infantile spasms), severe myoclonic epilepsy of Dravet-Dalla Bernardina and the Lennox-Gastaut syndrome, briefly describing their electroclinical semiology and therapeutic strategies using the range of available drugs, together with other medical and surgical therapeutic options. CONCLUSIONS: Treatment of severe epileptic encephalopathies of childhood is useless/not viable in neonatal myoclonic encephalopathies and severe myoclonic epilepsy of childhood, whilst in the West and Lennox-Gastaut syndromes the results depend on the aetiology, and are better in the cryptogenic types with control of 20% and 30% of the cases but with normal intelligence in only 5% and 9%. PMID- 11424040 TI - [Differential diagnosis of post-concussive syndrome]. AB - INTRODUCTION: Posconcussional syndrome is characterized by a heterogeneous group of somatic, cognitive and psychosocial symptoms, which occur in patients with head trauma, generally of mild severity. It is the neuropsychiatric postraumatic disorder more prevalent in the field of forensic medicine. DEVELOPMENT: Classical authors (Lishamn and Barraquer, for example) focused on controversial aspects of this syndrome, such as conceptual problems and etiology (organic versus functional). The objective of this report is to review the posconcussional literature in search of relevant aspects in forensic neuropsychology: conceptual aspects, epidemiology, etiology, clinical features, methodology for assessment, and its differential diagnosis with other postraumatic disorders, such as postraumatic stress disorder, adjustment disorder, anxiety disorder, mood disorders (major depressive disorder), substancerelated disorders, dementia due to head trauma, amnesic disorder, somatoform disorders, factitious disorder, malingering, chronic pain and chronic whiplash syndrome. PMID- 11424041 TI - [High doses of interferon in the treatment of multiple sclerosis]. PMID- 11424042 TI - [Immune events in central nervous system of early and late onset Alzheimer's disease patients]. AB - INTRODUCTION: Alzheimer s disease (AD) is a progressive degenerative disease affecting a significant proportion of the elderly population. The disease is characterized clinically by a progressive loss of memory function and mental impairment associated with the presence of degenerative well known pathological lesions. Although, the pathogenesis of AD is unclear; several reports indicate the involvement of immune factors. PATIENTS AND METHODS: This paper evaluates some cerebrospinal fluid immune markers from 21 patients with early and late AD and 20 age matched non-demented subjects. The analytical method included the evaluation of T cell subpopulations (using AcMc CD2, CD4, CD8) and activated T cells (AcMc HLA-DR and CD25) from CSF and peripheral blood by immunocytochemical techniques on a fixed cell slide as described by Bernd. The lymphocyte phenotype expressed as a percentage of positively stained cells for each cell surface marker evaluated. RESULTS: Some significant differences were observed for T cell subpopulations from different compartments, between the different AD groups and the controls (p< 0.05). Nevertheless, the most significant differences were found in the activated T cells from cerebrospinal fluid between AD groups and controls (p< 0.01). CONCLUSIONS: These results support the theory of neuroimmune dysregulation, probably involved in the progressive neurodegeneration and dementia in some AD. PMID- 11424043 TI - [Dietary cholesterol modifies pyroglutamyl aminopeptidase activity in mouse frontal cortex. Sexual differences]. AB - INTRODUCTION: Pyroglutamyl aminopeptidase (pGluAP) is an omega peptidase widely distributed in fluid and tissues, which hydrolyses biological active peptides including thyrotropin-releasing-hormone (TRH). In the last years, different endocrine and extraendocrine functions have been attributed to TRH. OBJECTIVES: The aim of present work is to study the influence of high dietary cholesterol on soluble and membrane bound pGluAP activity in frontal cortex of male and female mice. Material and methods. Soluble and membrane bound pGluAP activities of frontal cortex of mice feeding during 15 days, a standard diet enriched with cholesterol (1%) and cholic acid (0.5%) were measured using fluorimetric assays. RESULTS: Significant increases were observed in membrane bound pGluAP activity in males and females. No changes were detected in soluble pGluAP activity in frontal cortex of females but a significant increase was observed in this enzymatic activity in males. CONCLUSIONS: High dietary cholesterol induces a significant increase in tissue pGluAP activity. These increases may produce changes in TRH metabolism which may be related with alterations in its neuromodulatory functions and with the possible relationship between TRH and other neurotransmitter systems. PMID- 11424044 TI - [Findings on magnetic resonance in spastic cerebral palsy]. AB - OBJECTIVES: To study the types of lesion found on magnetic resonance (MR) in cerebral palsy (CP) and their correlation with a the type of CP, gestational age at birth and aetiology. PATIENTS AND METHODS: We made a retrospective review of all patients over one year in age, diagnosed as having spastic CP, who were seen in a paediatric neurology clinic between 1987 and 1998. The MR were evaluated blindly using data from the clinical history. RESULTS: Seventy two patients fulfilled criteria for inclusion in the study. The following results were obtained (all associations were statistically significant with p<0.01):1. Patterns of lesion: MR findings were classified into one of the following patterns: parasagittal lesion (7), diffuse multicystic encephalomalacia (4), periventricular leukomalacia (PVL) (26), bilateral dilatation of the basal ventricles without alteration of the signal from the periventricular white matter (5), bilateral lesions in the basal ganglia (5), focal multicystic encephalomalacia (limited to one vascular area) (4), unilateral lesions of hypersignal in the white matter (4), unilateral dilation of a lateral ventricle with or without homolateral cortical atrophy (3), malformations/disorders of migration (7), normal (7). 2. Correlation of the findings with the gestational age and type of CP: a) Spastic tetraplegia: 64% of the premature babies showed PVL. In newborn babies at term the lesions were more varied. b) Spastic diplegia: 80% of the premature babies had PVL and 83% of newborn babies at term had normal MR. c) Spastic hemiplegia: 81% had unilateral lesions. 3. Correlation with the aetiology: 62% of the cases of parasagittal lesions, diffuse multicystic encephalomalacia and bilateral lesions of the basal ganglia were due to hypoxic ischemic encephalopathy. CONCLUSION: The MR findings in CP show good correlation with gestational age, type of CP and aetiology. PMID- 11424045 TI - [Distal aneurysms of the vertebrobasilar system. A report of five cases]. AB - INTRODUCTION: Aneurysms distal to the circle of Willis are uncommon. Among the rarest of such aneurysms are those found in the territory of the vertebrobasilar artery. OBJECTIVE: To describe clinico-radiological aspects and the clinico surgical evolution of five patients with distal aneurysms of the cerebellar arteries. PATIENTS AND METHODS: Five patients (2.5%), with aneurysms of the cerebellar arteries, of a total of 194 patients with cerebral aneurysms admitted to our hospital between 1995 and 1998. RESULTS: All patients presented with subarachnoid hemorrhage. One patient also had an arteriovenous malformation (AVM). One patient refused surgical treatment, three were treated surgically and one using the endovascular approach. The patient who was successfully treated using the endovascular approach to an aneurysm of the antero-inferior cerebellar artery, and who also had an AVM had a further episode of bleeding following partial embolization of his AVM. The patients who were treated surgically and the one who refused surgical treatment made good progress. CONCLUSIONS: A multidisciplinary approach to the treatment of cerebral aneurysms is essential. Microsurgical techniques are very useful, especially in peripheral aneurysms. PMID- 11424046 TI - [Complications of stereotaxic surgery for cerebral tumors. Minimal access neurosurgery]. AB - INTRODUCTION: Complications of neurosurgery include the clinical, neurological and neurosurgical aspects. Their prevention and correction depend on satisfactory preoperative assessment and close postoperative follow-up. Although minimum access neurosurgery reduces some problems, the complexity and depth of many cerebral lesions cause problems. Therefore it is important to adhere to the above principles to obtain good results. OBJECTIVES: To determine the clinical complications, their early detection and course to be followed when they occur. PATIENTS AND METHODS: We studied 29 patients with intracranial tumors operated on using stereotaxic surgery during a period of two years. They were evaluated before and after surgery and the complications recorded. RESULTS: We discuss the most significant clinical aspects of peri-operative management. The main complications found were lower respiratory tract infection and hyperglycemia. CONCLUSIONS: These results show the need for suitable management, even in minimum access surgery, in patients with cerebral tumors operated on using stereotaxis and underline the most significant complications. PMID- 11424047 TI - [Executive dysfunction in Huntington's disease]. AB - INTRODUCTION: In the early stages of Huntington s disease (HD), there is selective vulnerability of the medium-sized spinous cells of the striate nucleus. These cells are connected with the dorsolateral prefrontal cortex, which is an area involved in cognitive processes concerning attention and executive function, forming part of one of the cortico-striatal circuits. OBJECTIVE: To analyze and interpret the executive dysfunction of a group of patients with HD at a moderately advanced stage. PATIENTS AND METHODS: We evaluated 16 patients and controls. Three clinical scales were used: the motor section of the UHDRS, the independence scale and the total functional capacity scale. For cognitive evaluation we used the study protocol of the Core Assessment Program for Intracerebral Transplantation for HD (CAPIT-HD). RESULTS: The patients with EH showed significant deterioration on the tests to evaluate cognitive function, attention and working memory. There were only two exceptions, the WEST and the direct order digit span. CONCLUSIONS: Patients with HD have executive dysfunction, probably due to dysfunction of the cortico-striatal circuits. We therefore agree with the proposal that fronto-striatal cognitive dysfunction is the most suitable name for the cognitive deterioration seen in this condition. PMID- 11424048 TI - [Characterization of the patients with cerebral infarcts discharged from a hospital in La Havana in 1998]. AB - OBJECTIVE: We made a study of the 79 patients who were discharged from the Instituto Nacional de Neurologia (La Havana, Cuba) during 1998, with the diagnosis of cerebral infarct, with the objective of analysing their behavior. PATIENTS AND METHODS: In this analysis we considered: the previous clinical history of factors and illnesses of risk, the certainty of the diagnosis in the emergency room, presenting symptoms, clinical signs, vascular territory, etiopathogenic category and results of cranial computerized axial tomography (CAT). RESULTS AND CONCLUSIONS: Arterial hypertension, smoking, cerebrovascular disease and ischemic cardiopathy were the commonest clinical features seen. There was close agreement between the diagnosis made in the emergency room and the final diagnosis. The presenting symptoms and clinical signs found in these patients were related to the vascular territory and motor involvement was the most constant. There was predominance of the carotid territory, especially of the left side. We also found predominance of atherothrombotic etiopathogenesis, with fewer cardio-embolic or unknown causes. There were few abnormal cranial CAT findings in patients with lacunar infarcts or infarcts localized to the posterior territory. PMID- 11424049 TI - [Prader-Willi syndrome large deletion on two brothers. Is this the exception that confirm the rule?]. AB - INTRODUCTION: Prader-Willi syndrome (PWS), a neuroendocrine disorder could be due to: a large paternally derived chromosome deletion of 15q11-13, to maternal uniparental disomy (UPD), or imprinting mutation (IC); amongst this last group five families, with inherited microdeletion encompassing SNRPN were described; in these families excluded a typical large deletion. Families with more than a child with PWS by classic large deletion have not been published. CLINICAL CASES: We report on a family with three children, 2 of which had typical clinical findings of PWS: mental retardation, hypogonadism, hypotonia, hyperphagia, obesity and also strabismus and synophridia; during pregnancy reduced fetal movement was noted. The Fish probes (SNRPN and D15S10), Methylation specific PCR (MPCR), Southern blot and microsatellite markers confirmed in the PWS brothers a large deletion at least of the area comprising between D15S63 and GABRA5. CONCLUSIONS: No previously described cases in the literature reviewed show for PWS brothers due to a classical deletion. Some possible reasons recurrence in this family could be: at random, germinal mosaicism, or abnormalities at gonadal level environmental factors such as hydrocarbon exposing occupations in fathers of PWS patients as has been referred to by different authors. The latter might be an explanation as the father was working from age of 17 and for over 12 years with paints at shipyards exposed to hydrocarbon and others mutagenic substances. We consider it would be important to bear this case in mind when giving genetic counseling. PMID- 11424050 TI - [Infantile bilateral striate necrosis]. AB - INTRODUCTION: Necrosis of the basal ganglia (NBG) is an uncommon condition in childhood. Cases of NBG have been reported in connection with metabolic disorders, infections, degenerative conditions, intoxications, head injuries and hypoxic-ischemic encephalopathies. The commonest clinical features include alteration of consciousness, chorea-athetoic or dystonic movements, spasticity, contractures of the limbs and convulsions. CLINICAL CASE: We describe the case of an eight year old girl, who after having clinical gastroenteritis, developed acute neurological dysfunction, associated with images on cranial computerized axial tomography (CAT) showing a possible intracranial tumour and magnetic resonance (MR) studies showing bilateral hypodensity of the basal ganglia. Three months later magnetic resonance showed that these lesions had almost completely disappeared. This supports the diagnosis of post-infectious lesions. CONCLUSIONS: The clinical course is very variable. Cases with a previous history of infection have a better prognosis. In our patient treatment was started with corticosteroids. She made a good recovery and was sent home with no sequelae five days later. The new imaging techniques (CAT and MR) have led to more cases of selective involvement of the basal ganglia being diagnosed. MR is the most specific technique for diagnosis in these patients. Although there is no effective specific treatment, treatment with biperidine, thyrotropic hormone and corticosteroids have been used with no apparent effect on the course of the disorder. PMID- 11424051 TI - [Intra-arterial fibrinolysis in vertebrobasilar system]. AB - INTRODUCTION: The acute vertebrobasilar occlussion is usually a life-treatening disease leading to death or major disability. The treatment with heparin and the selective fibrinolysis no show good results, for this reason the local intra arterial fibrinolysis appear as the choice treatment in patients with stroke and a agiography with basilar artery occlussion or intracranial vertebral artery occlusion. This tecnique has been proved to be effective treatment for selected patients with acute thromboembolic occlusion of the middle cerebral artery, ophthalmic artery and vertebrobasilar system, reducing the mortality in the vertebrobasilar system from 90% to 40%. CLINICAL CASE: We present a 63 years-old man with a vertebrobasilar thrombosis of a probable cardioembolic origen. He was treated with r-TPA local intra-arterial fibrinolysis, to get a recanalization of vertebrobasilar system. In the control TC we see a haemorragic sufusion in the brain stem. The follow-up see a patient with tetraparesis and palsy of the low cranial nerves and normal superior cerebral functions. CONCLUSIONS: The local intra-arterial fibrinolysis is the choise treatment in the vertebrobasilar thrombosis because the high morbimortality of this patology and the inefficacy of the others therapeutics. The result depend of many factors as the thrombo location, the neurologic state, the evolution time, the start of treatment, the colateral circulation, the nervous tissue reserve, etc, that have dificult predict the result, but it is best of the natural history of the disease. Is necesary, change the concept of emergency and the attitude front the isquemic cerebral disease at the sanitary leaders, the doctors, and the general population, for dispose of more means to cofront this pathology, which permit diminish the morbimortality and reduce the grade of incapacity. PMID- 11424052 TI - [Astrocytoma of the cerebellum and adverse effects of radiotherapy. A case report]. AB - INTRODUCTION: Although the nervous system is highly resistant to radiations, some cases have been reported about adverse effects of radiotherapy on patients with brain tumors. The following delayed effects have been observed: coagulation necrosis of all the cellular elements, fibrinoid and hyaline changes on the walls of the neoformed blood vessels, together with endothelial proliferation and perivascular fibrosis, reactive astrocytes, some of them with bizarre nuclei, as well as telangiectasia, hemorrhage and hemorrhagic infarcts. Telangiectasia and associated hemorrhage may occur in any of the irradiated zones of the brain. CLINICAL CASE: The present case is an example in which the indication of radiotherapy was due to an anatomopathological overdiagnosis. The first biopsy was diagnosed as a grade II cerebellar astrocytoma, after which 20 radiotherapy sessions were indicated. Nine years after that, a second operation was performed. This second biopsy revealed areas of fibrosis, hemorrhages, calcifications, necrosis, and thick hyaline-walled blood vessels. No tumor infiltration was observed. After an unfavorable evolution, the patient died. The microscopical study of the autopsy material revealed an area of neoplastic infiltration which was well defined as a pylocitic astrocytoma of the cerebellum. The rest of the cerebellum sections evidenced damage corresponding to an adverse effect of radiotherapy, particularly large zones of telangiectasia. CONCLUSIONS: It is stated that the patient s death was not because of the tumor. The adverse effects of radiotherapy, interpreted as tumor remains, motivated the second operation that generated complications causing the patient s death. PMID- 11424053 TI - [Bioethical considerations in the approach to patients with amyotrophic lateral sclerosis]. AB - INTRODUCTION: The traditional doctor-patient relation has become a great bioethical challenge due to the advances in science in recent years. This is particularly true when patients suffer diseases such as amyotrophic lateral sclerosis (ALS), a neurodegenerative disease with a relentless course and in spite of modern treatment 50% of the patients die within three years of first having symptoms of the disease. It therefore causes great psychological and social impact. OBJECTIVE: To analyze the great bioethical challenge which arises when diagnosing and treating a patient with ALS. DEVELOPMENT: In this paper we analyze the doctor-patient relationship, the principles of doing no harm and of being beneficial, and more modern concepts such as informed consent, biomedical investigations and euthanasia, as well as the importance of palliative medicine and rehabilitation to alleviate suffering and improve quality of life. Biomedical investigations should conform to the relevant national and international rules. We discuss the right of patients to be given truthful information. CONCLUSIONS: We recommend better training of doctors in all aspects of attention to these patients, with emphasis on the diagnosis and importance of rehabilitation, palliative medicine and the management of psychological aspects. Biomedical investigations should fulfil current regulations. We recommend discretion, complete or partial, with regard to information given to the patients and their relatives so as not to cause despair. PMID- 11424054 TI - [Juvenile myoclonic epilepsy]. AB - INTRODUCTION: In this paper we review the history of juvenile myoclonic epilepsy (JME) since the description by Herpin in 1852 and 1857 of the seizures of some epileptic patients which he called impulsions . Credit is due to Janz and Christian who in 1957 gave a detailed description of the condition which they called impulsive Petit mal and we know today as JME. Delgado-Escueta and his group in Los Angeles discovered the genetic basis of JME in some patients. DEVELOPMENT AND CONCLUSIONS: The homogeneity of the clinical and neurophysiological characteristics of series of patients with JME reported from many different parts of the work is striking. Diagnosis of JME is easy when the syndrome is known. However, in some patients a broad differential diagnosis should be considered as discussed in this article. The treatment of choice is still valproic acid as monotherapy. When this is not effective or side-effects occur, other useful drugs are available such as primadone. Of the other new antiepileptic drugs lamotrigine has been shown to be effective and good results have been obtained with topiramate, so both these drugs should be considered in some patients. Although response to treatment is excellent, a high incidence of relapse was seen in all studies when the patients stopped their treatment. PMID- 11424055 TI - [Partial seizures in childhood]. AB - INTRODUCTION AND OBJECTIVE: Partial seizures make up 49.3% of the epilepsies of school-age children and adolescents, and is the most prevalent type of epilepsy. Patients with complex partial seizures have more recurrences than other patients who have generalized seizures. We analyze the factors which worsen the prognosis in refractory partial epilepsies, their cost in childhood, the characteristics of the new drugs, and the few comparative studies of combinations of these drugs in resistant partial crises of childhood. DEVELOPMENT: The essential factors for prediction are previous brain damage, early onset of the seizures and presence on the EEG recording of marked paroxystic anomalies. Convincing studies on the economic aspects of the epilepsies should not be limited to the cost of the disease, but be descriptive studies which measure all the costs caused by epilepsy. It has been shown that the economic impact of the disorder on children with refractory epilepsy is 2.7 times the average cost of children whose seizures are well-controlled. There are no marked differences between the new antiepileptic drugs in the treatment of seizures of this type. Their appearance has led to reconsideration of what is rational bitherapy and what benefit may be obtained with it. CONCLUSIONS: When possible associations of antiepileptic drugs are considered, one should take into account criteria of efficacy and toxicity, the pharmacodynamic characteristics of the drugs to be used together and their pharmacokinetic interactions. It is necessary to avoid drugs with a low therapeutic index to which tolerance may develop or which interact frequently, and antiepileptic drugs with a broad spectrum, high therapeutic index and few pharmacokinetic interactions should be preferred. The association of valproate and viagabatrin may be very beneficial, since it has been suggested that there is a synergistic reaction in patients with partial seizures that are resistant to other drugs. The efficacy of the association of valproate and viagabatrin has been shown in patients with refractory partial seizures, since they have different modes of action and different spectrums, different profiles of toxicity and no pharmacokinetic interactions. Finally, we describe the basic principles of treatment of refractory partial seizures of childhood. PMID- 11424056 TI - [Why do the aphasic patients produce semantic errors?]. AB - INTRODUCTION: One of the most striking and surprising symptoms of the aphasic patients is the production of semantic errors. This kind of errors, which consists of replacing a word by another with a different but similar meaning, may appear when speaking, as it happens when a patient say dog in reference to a cat (semantic paraphasia), when reading as it happens when a patient reads dog where is writing cat (semantic paralexia), as well as in repetition or in writing. DEVELOPMENT: Some patients make these errors only in a specific linguistic modality (deep dyslexic patients make semantic errors in reading, deep dysphasic patients in repetition, etc.), but some patients make semantic errors in several modalities or even in all of them. Why these differences and what are the causes of these errors? Nowadays, models of linguistic processing built up from experiments with normal subjects provide us an answer to most of the above questions. From these models, by using appropriate tasks, it is possible to examine the reasons because of a patient makes certain errors. In fact, the results in different tasks help us to locate the deficit of each patient more than errors per se. It has been shown that sometimes the same type of error (e.g. a semantic paraphasia) can be caused by different processes in different patients. In this paper we will try to analyze which are the cognitive processes underlying semantic errors in each of the linguistic activities, when such processes are altered by injury, as well as to determine the best procedures to know what mechanisms are generating semantic errors in each aphasic patient. PMID- 11424057 TI - [Psychology, audition and language in different disorders of childhood. Communication and neuropsychological aspects]. AB - INTRODUCTION: A great number of language problems in children are related to neurological lesions in the central nervus systems. DEVELOPMENT: Cerebral palsy is one of the lesions more frequent. In many occasions the neurological alteration is accompanied by transmission hypoacusies, language delay an dysarthric problems in speech and voice. Moreover some of these cases have difficulties in attention, slow in speed of processing information, and different executive capacity depending on neurological and context factors. This work focuses into the aspects to consider the observation, evolution and treatment of these patients. PMID- 11424058 TI - [Aphasia: semiology and clinical patterns]. AB - INTRODUCTION: Human language is the final result of a group of brain activities directed towards the reception, integration and elaboration of linguistic messages. DEVELOPMENT: The disorganization of these activities following brain damage brings about a new linguistic functional level which constitutes the aphasic syndrome. Its assessment provides the clinician with information on the destroyed and impaired aspects and on those that have remained intact. The rehabilitation of aphasia should take into account the different components of the neurolinguistic semiology which have been evidenced by previous assessment, without forgetting that the rehabilitation process does not only have to deal with the expressive and receptive aspects of language, but also with a range of other possible cognitive impairments caused by the cerebral lesion and with the social and emotional problems that follow the new personal and social situation after the lesion. PMID- 11424059 TI - [Maintenance of logopedic orientation in a patient with oropharyngeal dysphagia of neurogenic origin]. AB - INTRODUCTION: Speech therapy treatment on patients who have a neurogenically induced dysphagia begins at hospital, in the intensive rehabilitation units. This study shows some of the difficulties found dealing with and orienting the family and the patient from a therapy point of view, mainly during his return to home surroundings. DEVELOPMENT: To better understand the dysphagic situation, normal swallowing steps (oral, pharyngeal and esophageal) steps have been described. These were then related to the dysphagic problems, with their evaluation, classification and characteristics. Under dysphagia rehabilitation the therapeutic guidance should be lead by the appropriate nutritional demands, and prevent aspirations. Some of the therapeutic procedures for treating and controlling neurogenic dysphagia have been described (head posture control, tactile and thermal stimuli, etc.). The author questions the difficulties found controlling orientation and maintaining conduct outside the hospital environment, principally with patients at risk from aspirations. PMID- 11424060 TI - [Luis Simarro: precursor of Spanish Neurology and Grand Master of the freemasons]. AB - Luis Simarro is considered to be the great precursor of Spanish Neurology, Neurohistology and Psychology. Together with Cajal he was one of the most outstanding figures of the Spanish freemasons in the field of medicine, where he reached the highest rank: Grand Master of the Spanish Great Orient. These historical notes summarize the scientific contribution of Simarro and his extensive work in the field of politics where he was influential in the modernization of Spain. PMID- 11424061 TI - [Eosinophilic meningoencephalitis in Cuba]. PMID- 11424062 TI - [Neurocysticercosis and epilepsy in Cuba]. PMID- 11424071 TI - Mitochondrial DNA haplogroups in Amerindian populations from the Gran Chaco. AB - Mitochondrial DNA from 141 individuals was typed for diagnostic restriction sites and the 9-bp region V deletion to examine the distribution of the founding mtDNA lineage haplotypes in three Amerindian populations (Mataco, Toba, and Pilaga) who currently inhabit the Argentinian part of the Gran Chaco. All four lineages were identified in the three tribes and four population samples studied. Disregarding ethnic or geographic origin, haplogroups B and D exhibit high incidence among the Gran Chaco inhabitants, whereas haplogroups A and C are present in a lower frequency. Three individuals possess none of the characteristic markers and, therefore, could not be assigned to one of those lineages. A neighbor-joining representation of F(ST) distances reflects the current geographic location of the populations, and this also corresponds to their historic distribution. After separating South America into four major regions (Tropical Forest, Andes, Gran Chaco, and Patagonia-Tierra del Fuego), the Gran Chaco populations present the highest average intragroup variability (Hs = 0.64) as well as the lowest intergroup diversity (G(')(ST) = 0.06). These findings suggest high levels of gene flow among the Chaco tribes, as well as with neighbor populations from outside the region. PMID- 11424072 TI - Infant feeding and weaning practices in Roman Egypt. AB - Current knowledge of infant feeding and weaning practices during the Roman period in Egypt is limited to scanty documentary and iconographic evidence. Stable nitrogen and carbon isotope analysis provides another avenue to explore this question. A sample of 49 infant and juvenile human skeletal remains from the Kellis 2 cemetery in the Dakhleh Oasis, Egypt, was used to determine patterns of infant feeding and weaning. delta(15)N values indicate that supplementary foods were introduced at around 6 months of age, and that weaning was complete by 3 years of age. By 6 months of age, delta(13)C values become increasingly enriched over adult values, and reach peak enrichment at approximately 1.5 years of age. Beyond this age, delta(13)C gradually declines to approach adult values. This enrichment in infant delta(13)C values is indicative of consumption of (13)C enriched supplementary foods. Based on isotopic study of faunal and botanical remains from the ancient village of Kellis, we conclude that at approximately 6 months of age, infants were fed milk of goat and/or cow. PMID- 11424073 TI - Social "status" and biological "status": a comparison of grave goods and skeletal indicators from Pontecagnano. AB - Comparison of funerary treatment and skeletal biology can be very informative about the interplay of social status and meanings and actual life conditions in ancient communities, but such comparison is rarely done, due in part to the disciplinary separation of bioanthropology and social archaeology in many archaeological traditions. In this paper, we analyze relations between skeletal pathologies and grave goods in a sample of 94 individuals from Pontecagnano (Salerno, Italy, seventh-third centuries BC). The results show that the relationship between health, activity, and social status as expressed in grave goods was complex. Some biological indicators considered typical of "stress" or biological status (enamel hypoplasia, cribra orbitalia, adult stature) bore no relation to social status. Other indicators, particularly those of activity and stress in adult life (trauma, Schmorl's nodes, periostitis), covaried with grave assemblage and help to outline a possible division of labor. As this analysis shows, when skeletal and archaeological data are used in conjunction, the result is a deeper picture of the social and economic life of the community than can be obtained from either source. PMID- 11424074 TI - Tissue contributions to sex and race: differences in tooth crown size of deciduous molars. AB - This study describes size of constituent deciduous tooth crown components (enamel, dentine, and pulp) to address the manner in which males characteristically have larger teeth than females, and the observation that teeth of American blacks are larger than those of American whites. Measurements were collected (n = 333 individuals) from bitewing radiographs using computer-aided image analysis. Tissue thicknesses (enamel, dentine, pulp) were measured at the crown's mesial and distal heights of contour. Deciduous mesiodistal molar crown length is composed of about 1/7 enamel, 1/3 dentine, and 1/2 pulp. Details differ by tooth type, but males typically have significantly larger dentine and pulp dimensions than females; there is no sexual dimorphism in marginal enamel thickness. Males scale isometrically with females for all variables tested here. Blacks significantly exceed whites in size of all tissues, but tissue types scale isometrically with blacks and whites with one exception: enamel thickness is disproportionately thick in blacks. While the absolute difference is small (5.56 mm of enamel in blacks summed over all four deciduous molar tooth types vs. 5.04 mm in whites), the statistical difference is considerable (P < 0.001). Aside from enamel, crown size in blacks is increased proportionately vis-a-vis whites. Principal components analysis confirmed these univariate relationships and emphasizes the statistical independence of crown component thicknesses, which is in keeping with the sequential growth and separate embryonic origins of the tissues contributing to a tooth crown. Results direct attention to the rates of enamel and dentine deposition (of which little is known), since the literature suggests that blacks (with larger crowns and thicker enamel) spend less time in tooth formation than whites. PMID- 11424075 TI - Cranial deformation in a subadult sample from Degirmentepe (Chalcolithic, Turkey). AB - Human skeletal remains from the Chalcolithic period of Degirmentepe, Turkey, near the Euphrates River, display artificial cranial deformation of the circular type. Human skeletal remains belonging to 31 individual have been dated to the Chalcolithic period (second half of the fifth millennium BC, uncalibrated). The remains include foetuses, infants, children and juveniles; but no adult skeletons. Either the adults were buried elsewhere or flooding of the Euphrates dragged the adult remains away. Artificial cranial deformation was observed in 13 of the 31 subadults ranging in age from 1 month to 13-14 years. Degirmentepe villagers might have used one or two circular constricting bands with the intent of producing artificial head deformation. As far as Anatolia and the Near Eastern region is concerned, the widespread use of circular head deformation may be an indication of longer-distance trade and more need for ethnic markers. PMID- 11424076 TI - Arrested development of secondary sexual adornments in subordinate adult male mandrills (Mandrillus sphinx). AB - Previous studies of semifree-ranging mandrills identified two morphological and social variants of the adult male, based on behavioral and secondary sexual characteristics. "Fatted" males are social, with highly developed sex skin coloration, large testes, high plasma testosterone levels, and fat rumps; while "nonfatted" males are peripheral or solitary, with paler sex skin, smaller testes, lower plasma testosterone, and slimmer rumps. We present a detailed study of morphology and group association for 10 adult male mandrills, living in two semifree-ranging groups in Gabon, in order to relate differences between males to dominance rank. The results show that rather than existing as two distinct morphotypes, male mandrills represent a continuous spectrum of possibilities between social males with fully developed secondary sexual characteristics, and solitary males with muted secondary sexual characteristics. Alpha males (N = 2) had the highest testosterone levels, the most colorful sex skin, and the most active sternal glands, and were the only males to spend 100% of their time with the social group. Rank relationships between nonalpha males (N = 8) were not always clear, but all subordinate males had lower testosterone levels and less development of the secondary sexual adornments, and were less group-associated than alpha males. These findings suggest that only alpha males have sufficient testosterone to develop full secondary sexual characteristics, and we propose possible socioendocrine mechanisms underlying the suppression of testosterone and secondary sexual development in subordinate adults. We discuss differences in secondary sexual development in relation to reproductive strategies, and discuss the evolution of alternative reproductive morphs in primates. PMID- 11424077 TI - Telemetered electromyography of the supinators and pronators of the forearm in gibbons and chimpanzees: implications for the fundamental positional adaptation of hominoids. AB - Extant apes are similar to one another, and different from monkeys, in features granting them greater range of forearm rotation and greater size of the muscles that produce this motion. Although these traits may have been independently acquired by the various apes, the possibility arises that such features reflect adaptation to the stem behavior of the hominoid lineage. Anticipating that knowledge of forearm rotatory muscle recruitment during brachiation, vertical climbing, arm-hanging during feeding, and voluntary reaching might point to this stem behavior, we undertook telemetered electromyographic experiments on the supinator, pronator quadratus, ulnar head of pronator teres, and a variety of other upper limb muscles in two gibbons and four chimpanzees. The primary rotator muscles of the hominoid forearm were recruited at high levels in a variety of behaviors. As had been suspected by previous researchers, the supinator is usually active during the support phase of armswinging, but we observed numerous instances of this behavior during which the muscle was inactive. No other muscle took over its role. Kinetic analyses are required to determine how apes can execute body rotation of armswinging without active muscular effort. The one behavior that is common to most extant apes, is rare in monkeys, and which places a consistently great demand on the primary forearm rotatory muscles, is hang feeding. The muscles of the supporting limb are essential to properly position the body; those of the free limb are essential for grasping food. Since the greater range of forearm rotation characterizing apes is also best explained by adaptation to this behavior, we join previous authors who assert that it lies at the very origin of the Hominoidea. PMID- 11424078 TI - Ontogeny of canine dimorphism in extant hominoids. AB - Many behavioral and ecological factors influence the degree of expression of canine dimorphism for different reasons. Regardless of its socioecological importance, we know virtually nothing about the processes responsible for the development of canine dimorphism. Our aim here is to describe the developmental process(es) regulating canine dimorphism in extant hominoids, using histological markers of tooth growth. Teeth preserve a permanent record of their ontogeny in the form of short- and long-period incremental markings in both enamel and dentine. We selected 52 histological sections of sexed hominoid canine teeth from a total sample of 115, from which we calculated the time and rate of cuspal enamel formation and the rate at which ameloblasts differentiate along the future enamel-dentine junction (EDJ) to the end of crown formation. Thus, we were able to reconstruct longitudinal growth curves for height attainment in male and female hominoid canines. Male hominoids consistently take longer to form canine crowns than do females (although not significantly so for our sample of Homo). Male orangutans and gorillas occasionally take up to twice as long as females to complete enamel formation. The mean ranges of female canine crown formation times are similar in Pan, Gorilla, and Pongo. Interspecific differences between female Pan canine crown heights and those of Gorilla and Pongo, which are taller, result from differences in rates of growth. Differences in canine crown heights between male Pan and the taller, more dimorphic male Gorilla and Pongo canines result both from differences in total time taken to form enamel and from faster rates of growth in Gorilla and Pongo. Although modern human canines do not emerge as significantly dimorphic in this study, it is well-known that sexual dimorphism in canine crown height exists. Larger samples of sexed modern human canines are therefore needed to identify clearly what underlies this. PMID- 11424079 TI - Brief communication: Cranial reconstruction of Dryopithecus. AB - This paper presents the first three-dimensional reconstruction of the cranium of the European Miocene ape, Dryopithecus. The reconstruction is based on remains collected from Can Llobateres, Spain (CLI 18000; Dryopithecus laietanus), D. fontani (St. Gaudens), and D. brancoi (Kordos and Begun [1997] Am. J. Phys. Anthropol. 103:277-294). Previously unrecognized facial fragments from CLI 18000 were included in the reconstruction. The result shows that Dryopithecus had a relatively short face, with a rather short and well-rounded neurocranium. From the lateral perspective, the face is concave at midface, and on the cranium the temporal lines run parallel to each other without forming a sagittal crest. PMID- 11424080 TI - Brief communication: Choice of washing method of hair samples for trace element analysis in environmental studies. AB - Determination of inseparable exogenous and endogenous levels of toxic trace elements such as lead in human scalp hair has been used to understand and assess population exposure to such elements. For any such analysis, washing of the hair samples to remove superficial contamination is the foremost requirement. However, the use of such washing methods prior to elemental analysis removes minute quantities of trace elements from the hair strands. This degree of loss is dependent on the washing method employed. In the present study, the element lead has been determined in human scalp hair after three methods of hair washing. It is apparent that the nonionic detergent-acetone method is best suited for securing valid lead analysis results in human scalp hair. PMID- 11424081 TI - Sp1 and kruppel-like factor family of transcription factors in cell growth regulation and cancer. AB - The Sp/KLF family contains at least twenty identified members which include Sp1-4 and numerous kruppel-like factors. Members of the family bind with varying affinities to sequences designated as 'Sp1 sites' (e.g., GC-boxes, CACCC-boxes, and basic transcription elements). Family members have different transcriptional properties and can modulate each other's activity by a variety of mechanisms. Since cells can express multiple family members, Sp/KLF factors are likely to make up a transcriptional network through which gene expression can be fine tuned. 'Sp1 site'-dependent transcription can be growth-regulated, and the activity, expression, and/or post-translational modification of multiple family members is altered with cell growth. Furthermore, Sp/KLF factors are involved in many growth-related signal transduction pathways and their overexpression can have positive or negative effects on proliferation. In addition to growth control, Sp/KLF factors have been implicated in apoptosis and angiogenesis; thus, the family is involved in several aspects of tumorigenesis. Consistent with a role in cancer, Sp/KLF factors interact with oncogenes and tumor suppressors, they can be oncogenic themselves, and altered expression of family members has been detected in tumors. Effects of changes in Sp/KLF factors are context dependent and can appear contradictory. Since these factors act within a network, this diversity of effects may arise from differences in the expression profile of family members in various cells. Thus, it is likely that the properties of the overall network of Sp/KLF factors play a determining role in regulation of cell growth and tumor progression. PMID- 11424082 TI - Homeobox genes in normal and malignant cells. AB - Homeobox genes are transcription factors primarily involved in embryonic development. Several homeobox gene families have so far been identified: Hox, EMX, PAX, MSX as well as many isolated divergent homeobox genes. Among these, Hox genes are most intriguing for having a regulatory network structure organization. Recent indications suggest the involvement of homeobox genes in (i) crucial adult eukariotic cell functions and (ii) human diseases, spanning from diabetes to cancer. In this review we will discuss the mechanisms through which homeobox genes act, and will propose a model for the function of the Hox gene network as decoding system for achieving specific genetic programs. New technologies for whole-genome RNA expression will be crucial to evaluate the clinical relevance of homeobox genes in structural and metabolic diseases. PMID- 11424083 TI - beta-estradiol stimulation of DNA synthesis requires different PKC isoforms in HepG2 and MCF7 cells. AB - The role exerted by protein kinase C (PKC) on estrogen-induced DNA synthesis has been investigated in hepatic and mammary gland cells, HepG2 and MCF7. 17-beta estradiol stimulated DNA synthesis in HepG2 and MCF7 cells, maximal effect occurring at 10 nM. DNA synthesis stimulation was prevented by anti-estrogen ICI 182,780 and by inhibitor of PKC, Ro 31-8220. The rapid estradiol effects in MCF7 cells were determined by following the inositol trisphosphate (IP(3)) production and PKC-alpha membrane translocation. After estradiol treatment the increase of IP(3) production, prevented by anti-estrogen or by phospholipase C (PLC) inhibitor (neomycin), was present in MCF7 cells. In MDA cells, devoid of estrogen receptor, no effect was observed. The PKC-alpha presence on the membranes appeared unchanged in MCF7 cells. The PLC inhibitors, neomycin and U73,122, and PKC-alpha down regulator, phorbol 12-myristate 13-acetate (PMA), were able to prevent estradiol-induced DNA synthesis in hepatoma cells, but ineffective in mammary cells; wortmannin, an inhibitor of phosphoinositide 3-kinases (PI3-K), blocked DNA synthesis in both cell lines. These data show that beta-estradiol, via an estrogen receptor-mediated mechanism, activates more signal transduction pathways, and consequently different PKC isoforms in two responsive cell lines. In both cell lines PI3-K/PKC pathway is functional to the estrogen regulation of DNA synthesis, whereas in HepG2 cells the parallel involvement of the PLC/PKC alpha pathway is present. The reported results indicate that the DNA synthesis stimulation by beta-estradiol requires the estrogen receptor and utilises one or more activated pathways in dependence on the cell equipment. PMID- 11424084 TI - Heparan sulfate mimetics modulate calpain activity during rat Soleus muscle regeneration. AB - Skeletal muscle regenerates after injury. Tissue remodelling, which takes place during muscle regeneration, is a complex process involving proteolytic enzymes. It is inferred that micro and milli calpains are involved in the protein turnover and structural adaptation associated with muscle myolysis and reconstruction. Using a whole-crush injured skeletal muscle, we previously have shown that in vivo muscle treatment with synthetic heparan sulfate mimetics, called RGTAs (for ReGeneraTing Agents), greatly accelerates and improves muscle regeneration after crushing. This effect was particularly striking in the case of the slow muscle Soleus that otherwise would be atrophied. Therefore, we used this regeneration model to study milli and micro calpain expressions in the regenerating Soleus muscle and to address the question of a possible effect of RGTAs treatment on calpain levels. Micro and milli calpain contents increased by about five times to culminate at days 7 and 14 after crushing respectively, thus during the phases of fibre reconstruction and reinnervation. After 64 days of regeneration, muscles still displayed higher levels of both calpains than an intact uninjured muscle. Milli calpain detected by immunocytochemistry was shown in the cytoplasm whereas micro calpain was in both nuclei and cytoplasm in small myofibres but appeared almost exclusively in nuclei of more mature fibres. Interestingly, the treatment of muscles with RGTA highly reduced the increase of both milli and micro calpain contents in Soleus regenerating muscles. These results suggest that the improvement of muscle regeneration induced by RGTA may be partly mediated by minimising the consequences of calpain activity. PMID- 11424085 TI - Role of AKT kinase in sphingosine-induced apoptosis in human hepatoma cells. AB - Our previous work has shown that a number of sphingolipid metabolites including sphingosine, sphinganine, and other long-chain bases potently induced apoptosis in human hepatoma cells. In this study, we examined the possibility that sphingosine may trigger apoptosis in human hepatoma cells via inhibition of anti apoptotic pathways. We investigated the effect of sphingosine on AKT kinase, a serine/threonine kinase which was found to protect cells from apoptosis induced by a variety of extracellular stresses. Our results indicated that sphingosine inhibited basal and serum-stimulated AKT kinase activity in a dose-dependent manner in hepatoma cells. Additionally, sphingosine-induced inhibition of AKT kinase was correlated with induction of apoptosis in these cells. Pretreatment of insulin, a potent stimulator of AKT kinase, partially reversed the inhibition of AKT kinase by sphingosine and counteracted the apoptotic action of this sphingolipid. Expression of activated AKT kinase partially protected cells from sphingosine-induced apoptosis, whereas expression of kinase-dead AKT kinase had no effect. The molecular mechanism by which AKT kinase suppressed the apoptotic action of sphingosine was investigated. Our results showed that increased release of cytochrome C from mitochondria and subsequent activation of caspase-3 were detected in sphingosine-treated hepatoma cells. On the contrary, expression of activated AKT kinase in Hep3B cells attenuated cytochrome C release and caspase-3 activation induced by sphingosine. Taken together, these findings suggest that suppression of AKT kinase is one of the mechanisms by which sphingosine induces apoptosis in hepatoma cells and activation of AKT kinase may inhibit sphingosine induced apoptosis by blocking a step upstream of cytochrome C release and caspase 3 activation. PMID- 11424086 TI - Purification of activins from androgen-independent Shionogi carcinoma cells demonstrates enhanced expression of activin betaB-subunit under androgen-depleted cell conditions in vitro and in vivo. AB - Here, we report characterization of growth factors secreted from androgen independent mouse mammary Shionogi carcinoma cells. Previous isolation of fibroblast growth factor 8 (FGF8) from androgen-dependent Shionogi carcinoma SC-3 cells prompted us to characterize growth factors secreted from the androgen independent cells. After several purification procedures, mitogens for NIH3T3 cells from the androgen-independent cells were identified as activins on the grounds that activin betaA- and betaB-subunits are detected in the active fractions by Western blotting and that the growth-promoting effects by the active fractions are specifically inhibited in the presence of follistatin. In addition, exogenous activins, but not inhibin, stimulated the growth of NIH3T3 cells in a dose-dependent manner. Interestingly, transcripts of activin betaB-subunit were predominantly found in the androgen-independent cells while its betaA-subunit was universally expressed in both androgen-dependent and -independent Shionogi carcinoma cells. In concordant with this in vitro finding, transcripts of activin betaB-subunit were enhanced in murine prostates after castration. Therefore, expression of activin betaB-subunit, but not its betaA-subunit, is likely to be related with androgen-depleted cell conditions in prostates, and possibly in androgen-related cancers. PMID- 11424087 TI - Heterogeneity of the signal transduction pathways for VEGF-induced MAPKs activation in human vascular endothelial cells. AB - Vascular endothelial growth factor (VEGF) activates ERK and p38 MAPK in endothelial cells (ECs). The present study was aimed to compare its intracellular signal transduction pathways between three primary cultures of human ECs including human aortic ECs (HAECs), human umbilical vein ECs (HUVECs), and human microvascular ECs (HMVECs). VEGF activated ERK and p38 MAPK in all of three ECs. Isoforms of p38 MAPK that were activated by VEGF in HUVECs were p38-alpha and p38 delta. GF109203X, a specific inhibitor of PKC, markedly inhibited VEGF-induced activation of ERK and p38 MAPK in HAECs and HUVECs, whereas it exhibited little effect in HMVECs. In contrast, dominant negative mutant of Ha-Ras almost completely abrogated VEGF-induced activation of ERK and p38 MAPK in HMVECs. Although dominant negative mutant of Ha-Ras substantially inhibited the basal activities of ERK and p38 MAPK, it exhibited marginal effect on VEGF-induced activation of ERK and p38 MAPK in HUVECs and HAECs. The activation of Ras by VEGF appeared to be most prominent in HMVECs. These results indicate that intracellular signal transduction pathways for VEGF-induced activation of MAPKs are heterogeneous and vary depending on the origin of ECs. PMID- 11424088 TI - Fetal and adult human skin fibroblasts display intrinsic differences in contractile capacity. AB - One of the differences between fetal and adult skin healing is the unique ability of fetal wounds to heal without contracture and scar formation. Studies have shown that the ratio between the three isoforms of TGFbeta is different in adult and fetal wounds. Thus, we analyzed the capacity of adult and fetal human skin fibroblasts to contract collagen gels after stimulation with TGFbeta isoforms. In control medium, fetal fibroblasts had a contractile capacity similar to that of adult fibroblasts. However, the growth capacity of fetal fibroblasts was completely inhibited, in contrast to adult fibroblasts. When cells were treated with TGFbeta, fetal fibroblasts showed an inhibition of their contractile capacity whereas adult fibroblasts further contracted gels. The contractile response was similar for all isoforms of TGFbeta although TGFbeta3 always had the strongest effect. We considered that the regulation of cell contractile capacity by TGFbeta may be dependent on receptor expression for this cytokine, on myofibroblast differentiation of the cells, or in cell links with matrix. Since TGFbeta receptor analysis did not show differences in receptor affinity, we studied the expression of alpha-smooth muscle (SM) actin, a fibroblast contractile marker and of three integrins, the cell surface receptors specific of the attachment of the fibroblasts with collagen matrix. We observed that the expression of alpha-SM actin and alpha3 and beta1 integrin subunits was increased when TGFbeta was added to the medium of adult fibroblasts whereas the levels of the alpha1 and alpha2 subunits were unchanged. In contrast, fetal fibroblasts treated with TGFbeta showed a decrease of alpha1, alpha2, and beta1 integrin expression but no change in alpha3 integrin and in alpha-SM actin expression. These results indicate that intrinsic differences between fetal and adult fibroblasts might explain their opposite responses to TGFbeta stimuli. The variations in their alpha-SM actin and integrin expression patterns represent potentially important mechanisms used by fetal fibroblasts to regulate their response to cytokines, and likely contribute to the resultant differences in the quality of wound repair. PMID- 11424089 TI - Role of protein kinase Cdelta in transmitting hypoxia signal to HSF and HIF-1. AB - An hypoxic microenvironment is an important modulator of gene expression in many pathophysiological conditions. In this study, we show a coordinate activation of heat shock transcription factor (HSF) and hypoxia-inducible factor-1 (HIF-1) in RIF tumor cells by hypoxia. Since heat shock protein (hsp) and angiogenic factor genes that are regulated by HSF and HIF-1 are thought to contribute to the malignant progression of hypoxic tumor cells, it was of our major interest to identify the components that are responsible for the activation of both HSF and HIF-1. Our finding that a bioflavonoid quercetin (QCT), a well known inhibitor of hsp gene expression, significantly inhibited the transcriptional activation of HSF and HIF-1 strongly suggests that QCT-sensitive molecule(s) is involved in the transcriptional activation of HSF and HIF-1 by hypoxia. Our results revealed that PCKalpha, delta and epsilon isoforms are expressed in RIF cells, but only PKCdelta was specifically translocated to the membrane by hypoxia. Our results also revealed that the translocation of PKCdelta was completely abrogated by QCT. Moreover, inhibiting the PKCdelta activation, either pharmacologically with phorbol 12-myristate 13-acetate or with bisindolymaleimide II or genetically by transient transfection of a dominant negative PKCdelta, significantly inhibited the transcriptional activation of HSF and HIF-1 by hypoxia. These results strongly substantiate a view that the PKCdelta isozyme is the QCT-sensitive molecule that plays an important role in transmitting hypoxia signals to both HSF and HIF-1. Here we show that the membrane translocation of PKCdelta is dependent on the activation of phosphoinositol 3-kinase (PI3K). Treatment with PI3K inhibitor, wortmannin or LY294002, abrogated not only PKCdelta translocation but the subsequent transcriptional activation of HSF and HIF-1 by hypoxia. Together, our study shows that the PKCdelta isozyme acts as a shared component in transmitting hypoxia-induced signals to both HSF and HIF-1, and that the upstream regulator of PKCdelta is PI3K. PMID- 11424090 TI - Possible involvement of protein kinases and Smad2 signaling pathways on osteoclast differentiation enhanced by activin A. AB - Bone tissues reportedly contain considerable amounts of activin A and follistatin, an activin A-binding protein. In the present study, we found that follistatin strongly inhibited osteoclast formation in cocultures of mouse bone marrow cells and primary osteoblasts induced by 1alpha,25 dihydroxyvitamin D(3), prostaglandin E(2), and interleukin-1alpha. Antibody aganist activin A also inhibited the osteoclast formation. Furthermore, activin A synergistically stimulated osteoclast differentiation mediated by receptor activator NF-kappaB ligand (RANKL). RT-PCR analysis revealed that osteoblasts produced not only activin A but also follistatin. Western blot analysis of a panel of phosphorylated proteins revealed that activin A stimulated the phosphorylation of p44/42 mitogen activated protein (MAP) kinase (ERK1/2) and p38 MAP kinase in macrophage colony-stimulating factor-dependent bone marrow macrophages (M BMMPhis). In addition, phosphorylation of Smad2 was observed in M-BMMPhis stimulated with activin A. These findings indicate that the phosphorylation of p44/42 MAP kinase, p38 MAP kinase, and Smad2 is involved in activin A-enhanced osteoclast differentiation induced by RANKL. Taken together, these results suggest that both activin A and follistatin produced by osteoblasts may play an important role in osteoclast differentiation through MAP kinases and Smad2 signaling pathways. PMID- 11424091 TI - Role of transcription factor Ets-1 in the apoptosis of human vascular endothelial cells. AB - Transcription factor Ets-1 is induced in endothelial cells (ECs) by angiogenic factors, and promotes angiogenesis by inducing angiogenesis-related genes such as MMPs and integrin beta3. Here, we examined the effect of Ets-1 on apoptosis in ECs. Overexpression of Ets-1 in human umbilical vein endothelial cells (HUVECs) induced apoptosis under the serum-deprived condition. VEGF inhibited apoptosis and augmented the DNA binding of Ets-1 in HUVECs. The inhibition of transcriptional activity of endogenous Ets-1 by a dominant negative molecule intensified the anti-apoptotic effect of VEGF. Caspase inhibitors blocked apoptosis of HUVECs induced by Ets-1. DNA array analysis showed that Ets-1 up regulated pro-apoptotic genes such as Bid, cytochrome p450, caspase-4, p27, and p21 more than 2 fold, and down-regualted anti-apoptotic genes such as DAD-1, AXL, Cox-2, IAP-2, and MDM-2 less than 0.5 fold in HUVECs. These results indicate that Ets-1 itself is pro-apoptotic to ECs by modulating the expression of apoptosis related genes. PMID- 11424092 TI - Pigment epithelium-derived factor inhibits retinal and choroidal neovascularization. AB - In this study, we investigated whether overexpression of pigment epithelium derived factor (PEDF) by gene transfer can inhibit neovascularization by testing its effect in three different models of ocular neovascularization. Intravitreous injection of an adenoviral vector encoding PEDF resulted in expression of PEDF mRNA in the eye measured by RT-PCR and increased immunohistochemical staining for PEDF protein throughout the retina. In mice with laser-induced rupture of Bruch's membrane, choroidal neovascularization was significantly reduced after intravitreous injection of PEDF vector compared to injection of null vector or no injection. Subretinal injection of the PEDF vector resulted in prominent staining for PEDF in retinal pigmented epithelial cells and strong inhibition of choroidal neovascularization. In two models of retinal neovascularization (transgenic mice with increased expression of vascular endothelial growth factor (VEGF) in photoreceptors and mice with oxygen-induced ischemic retinopathy), intravitreous injection of null vector resulted in decreased neovascularization compared to no injection, but intravitreous injection of PEDF vector resulted in further inhibition of neovascularization that was statistically significant. These data suggest that sustained increased intraocular expression of PEDF by gene therapy might provide a promising approach for treatment of ocular neovascularization. PMID- 11424093 TI - Involvement of Smads in TGFbeta1-induced furin (fur) transcription. AB - Furin is recognized as being one of the main convertases of the cellular constitutive secretion pathway but the mechanisms regulating its expression are still unknown. We have previously demonstrated that TGFbeta1 up-regulates its own converting enzyme, furin, creating a novel activation/regulation cycle of potential importance in a variety of physiological and pathophysiological conditions. The fur (fes upstream region) gene is regulated via three alternative promoters; P1, P1A, and P1B. To gain insight into the molecular mechanism(s) underlying this up-regulation, we performed transient cell transfections with P1, P1A, and P1B promoter luciferase constructs. Transfection experiments in HepG2 cells revealed that fur P1 promoter is the strongest and the most sensitive to TGFbeta1 stimulation (5 ng/ml) (3.2-fold vs. 2.4-fold for P1A and 2.1-fold for P1B). Cotransfection with either a dominant negative mutant form of Smad2 [Smad2(3SA)] or a known Smad inhibitor [Smad7] inhibit constitutive and TGFbeta1 induced luciferase activity indicating the participation of endogenous Smads. Increased levels of TGFbeta1-induced transcriptional activation of the P1 promoter by overexpression of Smad2 and/or Smad4 is greatly reduced in the presence of Smad2(3SA) and completely inhibited by Smad7, suggesting the participation of endogenous Smad2/Smad4 complexes. Furthermore, the fork-head activin signal transducer (FAST-1), known to interact with Smad2/Smad4 complexes, is a potent stimulator of TGFbeta1-induced transactivation of the fur P1 promoter. Five prime-deletion analysis of this promoter identified the proximal region (between positions -8734 and -7925), as the nucleotide stretch that carries most of the transcriptional activation of fur P1 promoter by Smad2. Overall, the present data demonstrate that Smad2 and Smad4 possibly in complex with FAST-1 or other DNA binding partners participate in the constitutive and inducible transactivation of the fur P1 promoter. This represents the first detailed study of the transcriptional regulation of the fur gene. PMID- 11424094 TI - Cancer therapies: basic and clinical perspectives in brain, prostate, and lung tumors: Naples, September, 24-27, 2000. AB - A continuous flow of theoretical and practical information among basic research, diagnosis, and therapeutic innovation is a crucial process to achieve a timely and effective progress in defeating human cancer. According to this essential concept, the main objective of the Fourth Joint International Cancer Conference "Cancer Therapies: Basic and Clinical Perspectives in Brain, Prostate and Lung Cancer" has been of gathering together basic scientists and clinicians who represent scientific opinion leaders in their field, to present and discuss the most recent scientific achievements in basic and clinical perspectives, advanced diagnostic and therapeutic strategies, and molecular and cellular therapeutic approaches in brain, prostate, and lung cancer. PMID- 11424095 TI - Preoperative evaluation of periampullary tumors by endoscopic sonography, transabdominal sonography, and computed tomography. AB - PURPOSE: The purpose of this retrospective study was to compare the sensitivity of endoscopic sonography (EUS), transabdominal sonography (US), and CT in the detection of, local staging of, and prediction of vascular involvement by or distant metastasis from periampullary tumors. METHODS: Seventy-four consecutive patients with presumed periampullary tumors were evaluated by EUS, US, and CT during a 3.25-year period. The local staging accuracy of the modalities was assessed in the 36 patients with solid tumors who underwent surgery. The sensitivity of the modalities in predicting vascular involvement and distant metastasis was assessed in the 56 patients with carcinomas. RESULTS: EUS was the most sensitive modality in the detection (EUS, 97%; US, 24%; and CT, 39%; p < 0.001 for EUS versus US or CT) and T classification (EUS, 72%; US, 11%; CT, 22%; p < 0.001 for EUS versus US or CT) of periampullary tumors. EUS also had better sensitivity than US in detecting lymph node metastasis from periampullary cancers (EUS, 47%; US, 7%; and CT, 33%; p = 0.02 for EUS versus US; p = 0.7 for EUS versus CT). The accuracy of EUS in determining the T classification (without stent, 81%; with stent, 65%) and N classification (without stent, 80%; with stent, 70%) tended to decrease in the presence of an endobiliary stent, but the differences were not significant. EUS was the most sensitive modality in demonstrating vascular involvement (EUS, 100%; US, 0%; and CT, 33%; p = 0.002 for EUS versus US; p = 0.03 for EUS versus CT) but was not significantly different in detecting distant metastasis (EUS, 11%; US, 44%; and CT, 44%). CONCLUSIONS: EUS is superior to US and CT in the local assessment of periampullary tumors. The staging accuracy of EUS is minimally but not significantly affected by the presence of an endobiliary stent. PMID- 11424096 TI - Gallbladder wall thickening in mononucleosis syndromes. AB - PURPOSE: We used sonography to measure gallbladder wall thickness in patients with mononucleosis syndromes and then evaluated laboratory data, spleen size, and clinical evolution to assess any relationship between gallbladder wall thickening (GBWT) and the severity of disease. METHODS: We retrospectively reviewed the medical records, sonograms, and sonographic reports of 39 patients who were diagnosed with mononucleosis syndromes on the basis of fever, tonsillopharyngitis, cervical adenopathy, hepatosplenomegaly, and lymphocytosis with atypical lymphocytes. All 39 were included in the study. The gallbladder wall thickness in each patient was sonographically determined. GBWT was defined as a wall thickness exceeding 3 mm. We assessed the laboratory data and clinical evolution in each patient, and the differences between patients with and without GBWT were statistically analyzed. RESULTS: Six patients (15%) had GBWT. The mean atypical lymphocyte count +/- standard deviation (SD) in the patients with GBWT (1,830/microl +/- 1,000/microl) was significantly higher than that in patients without GBWT (1,140/microl +/- 660/microl; p < 0.05). The mean total serum protein and serum albumin levels in the patients with GBWT (6.6 mg/dl +/- 0.7 mg/dl and 3.7 mg/dl +/- 0.5 mg/dl, respectively) were significantly lower than those in patients without GBWT (7.3 mg/dl +/- 0.4 mg/dl and 4.1 mg/dl +/- 0.3 mg/dl, respectively; p < 0.05). The duration of hospitalization in the patients with GBWT (14 +/- 8.5 days) was significantly higher than that in patients without GBWT (8 +/- 3.5 days; p < 0.05). CONCLUSIONS: GBWT in mononucleosis syndromes may be a sign of the severity of the illness and when present indicates the need to carefully monitor the clinical course. PMID- 11424097 TI - Transabdominal sonography of the normal gastroesophageal junction in children. AB - PURPOSE: Because sonography identifies abnormalities of the gastroesophageal junction, it is essential to understand the normal sonographic anatomy. The aim of this study was to determine the normal sonographic appearance of the gastroesophageal junction and its variations and to provide measurements of the abdominal esophagus in asymptomatic, healthy children. METHODS: In this prospective study, 124 healthy children (75 boys and 49 girls), aged 2 days-12 years, underwent abdominal sonography. With the patient in a supine position, the transducer was placed under the xiphoid and the ultrasound beam was directed cephalad through the window of the left lobe of the liver. The length of the abdominal esophagus was measured from the point at which it penetrated the diaphragm to the gastroesophageal junction. The thickness was measured on the anterior wall at the midpoint of the abdominal esophagus. RESULTS: The gastroesophageal junction was identified by sonography in all of the children. The mean length of the abdominal portion of the esophagus ranged from 18 mm in the newborns to 34 mm in children older than 6 years. The wall thickness ranged from 2.4 mm to 5.7 mm. CONCLUSIONS: Our results indicate that visualization of the gastroesophageal junction and measurement of the abdominal esophagus are readily achievable with real-time sonography in healthy children. PMID- 11424098 TI - Comparison of the sonographic features of acalculous and calculous submandibular sialadenitis. AB - OBJECTIVE: The purpose of this study was to compare the sonographic features of acalculous and calculous sialadenitis of the submandibular gland. METHODS: A retrospective review of sonograms of the neck was performed in 25 patients with submandibular sialadenitis: 8 with acalculous and 17 with calculous disease. The submandibular glands were assessed for size, shape, border, and echogenicity; presence of dilated ducts or other intraglandular lesions; inflammatory changes in adjacent tissues; lymphadenopathy; and involvement of other salivary glands. RESULTS: In the group with acalculous sialadenitis, 4 (50%) of the 8 patients had unilateral disease, and 11 (92%) of 12 glands were rounded. In all cases (100%), multiple hypoechoic lesions were diffusely distributed throughout the submandibular glands against a heterogeneous parenchymal background. The lesions ranged from 3 to 15 mm and were oval or round. Confluent lesions were noted in 2 glands (17%). There was no sonographic evidence of duct dilatation, calculi, or abnormal lymph nodes. All patients with calculous sialadenitis had unilateral disease; 9 had a main duct calculus (53%), 7 had intraglandular calculi (41%), and 1 had both (6%). In 14 (82%) of 17 glands, normal shape was maintained, and 11 (65%) of 17 had duct dilatation. CONCLUSIONS: Acalculous submandibular sialadenitis differs from the calculous form of the disease. The former has characteristic sonographic features, including a round gland with numerous hypoechoic lesions in a heterogeneous parenchymal background. Sonographic imaging in conjunction with fine-needle aspiration is useful for detection of this disorder. PMID- 11424099 TI - Ultrasound-guided testicular sperm aspiration in azoospermic patients: a new sperm retrieval method for intracytoplasmic sperm injection. AB - PURPOSE: We investigated the technique of ultrasound-guided testicular sperm aspiration (USTSA) and compared it with "blind" testicular sperm aspiration (TSA) in patients with nonobstructive azoospermia. METHODS: Thirty-nine consecutive azoospermic men underwent TSA, 16 under sonographic guidance (USTSA group) and 23 with no imaging guidance (TSA group). Clinical and hormonal evaluation and sonography of the scrotum and testes were performed 1-2 days before the procedure. The aspiration was done using short-term general anesthesia. Follow-up consisted of sonographic reexamination of the scrotum and testes immediately and 1 month after the procedure. RESULTS: Intraoperative sonography with power Doppler imaging enabled good visualization of the testicular parenchyma, easy sampling, and avoidance of prominent vessels. Sufficient material was retrieved in 15 USTSA patients (94%) and 19 TSA patients (83%). No patients needed more than 4 hours' ambulatory hospitalization after the procedure. In the remaining 5 patients, aspiration failed to yield sperm, so open biopsy was performed. In those patients, postaspiration surgical exploration revealed subtunical bleeding in 3 patients after TSA but none after USTSA. Late minor complications occurred in 2 patients (13%) in the USTSA group and 7 (30%) in the TSA group. No difference was found between the 2 groups in pregnancy rate in the patients' female partners. CONCLUSIONS: USTSA is a safe and accurate method for sperm retrieval in azoospermic patients. PMID- 11424100 TI - Two small intrahepatic cholangiocarcinomas with different sonographic appearances. AB - We report on 2 cases of small intrahepatic (or peripheral) cholangiocarcinomas initially detected by sonography and completely resected. The lesions had a peripheral hypoechoic component and a central isoechoic or hyperechoic component. One had posterior acoustic shadowing on sonography, and the other did not. Comparison with pathologic findings in both cases indicated that severe fibrotic changes may have caused the posterior acoustic shadowing. This finding is extremely rare. We found only 1 previously reported case of cholangiocarcinoma accompanied by acoustic shadowing. PMID- 11424101 TI - Post-traumatic high-flow priapism: management by superselective transcatheter autologous clot embolization and duplex sonography-guided compression. AB - High-flow priapism is caused by uncontrolled arterial inflow into the penis, often from direct arterial trauma. We report a case of arterial or high-flow priapism that was treated with a combination of selective transcatheter autologous clot embolization and duplex sonography-guided compression therapy. Sonography showed a pulsatile, anechoic area at the base of the penis inside the left corpus cavernosum after blunt perineal trauma, and color Doppler examination revealed an arteriosinusoidal fistula. Autologous clot embolization was only partially successful, and 3 sessions of ultrasound-guided compression therapy were necessary to achieve complete thrombus formation in the fistulous tract, which became isoechoic a day after compression therapy. PMID- 11424102 TI - Cutaneous seeding after ultrasound-guided percutaneous ethanol injection for treatment of hepatocellular carcinoma. AB - Cutaneous seeding is a rare complication of interventional ultrasound procedures. We describe a case of needle-track cutaneous seeding of hepatocellular carcinoma (HCC) after sonographically guided percutaneous ethanol injection (PEI). In our case, the seeding might have been related to the type of needle used and the multiple passes required to treat the liver lesion. Despite the risk of needle track seeding, PEI remains useful in the treatment of HCC. PMID- 11424103 TI - Sonographic appearance of lipoma of the inferior vena cava. AB - Lipomas of the major central veins are rare, and their sonographic features have rarely been reported. We report a case in which a lipoma of the inferior vena cava (IVC) was incidentally identified during liver sonography in a 68-year-old man. Sonography of the upper abdomen revealed coarsening of the liver echotexture, with a relatively small liver and atrophy of the right hepatic lobe. A 2-cm hypoechoic hepatic nodule was visualized in segment IV. Sonography also showed a 2.2- x 1.8-cm, highly echogenic, homogeneous nodule in the hepatic segment of the IVC at its junction with the right atrium. The lumen of the IVC was partially occluded. The CT and MRI appearances of the IVC lesion were consistent with a lipoma, which was not treated because there was no clinical evidence of obstruction. Histopathologic analysis following a sonographically guided biopsy of the liver nodule revealed hepatocellular carcinoma, which was treated by percutaneous ethanol injection. PMID- 11424104 TI - Obstructive jaundice caused by hepatocellular carcinoma: detection by endoscopic sonography. AB - Tumor thrombus in the extrahepatic biliary tree is a rare mechanism of obstructive jaundice. We present a patient with a minute hepatocellular carcinoma in the caudate lobe that invaded the common hepatic duct and caused biliary obstruction. Endoscopic sonography showed a tumor thrombus with central echogenicity and a "nodule-in-nodule" pattern and suggested the correct diagnosis. PMID- 11424105 TI - Identification of key mutations in HIV reverse transcriptase gene can influence the clinical outcome of HAART. AB - Therapeutic failures due to in vivo loss of drug sensitivity are still a major problem in AIDS care. Currently, the role of and methods for detecting resistant mutant strains in patients before therapeutic choices are still under debate. To investigate the relevance of screening for key mutations alone the commercial INNO-LiPA HIV-1 RT method was applied retrospectively to analyzing several HIV codons correlated with resistance to RTI (reverse-transcriptase inhibitors) in sera from 62 patients before starting HAART protocols, selected on the basis of clinical parameters. INNO-LiPA detected several resistant mutant strains, which were strictly consistent with previous selective pressure in the patients. A significant correlation between genotype pattern and response to HAART was found. The presence of key mutations associated with resistance to one or two RTI included in the protocol correlated with a decrease in treatment benefits, whereas patients with wild-type or non-resistant viral strains exhibited better response to HAART. Even if this information had been available when treatment was started, 45 of the patients would not have received different treatment. When compared with the total number of patients, the subgroup receiving a treatment that was considered retrospectively as consistent with the key mutation pattern exhibited a significantly better outcome. Although the interpretation of resistance-related key mutations needs improvement, this surrogate LiPA method seems to maintain a predictive role in the management of HIV infection, and is less expensive. PMID- 11424106 TI - Antibodies raised to short synthetic peptides with sequences derived from HIV-1 SF2 gp120 can both neutralize and enhance HIV-1 SF13: a later variant isolated from the same host. AB - HIV-1 SF13 emerged in a patient with immunity to HIV-1 SF2. This study determined the effect of antibodies raised to HIV-1 SF2 on the replication of the later variant. Antisera in rats were raised previously to a complete set of overlapping, synthetic 15mer peptides following the sequence of HIV-1 SF2 gp120. These sera have now been used in neutralization and enhancement assays against viruses derived from molecular clones of both variants. The sets of peptides inducing neutralizing antibodies to the two variants overlap. Antibodies to the third variable region of HIV-1 SF2 only neutralize the homologous virus whereas those to the second and fourth variable regions neutralize both variants. In contrast, the sets of major epitopes involved in enhancement do not overlap. Epitopes for both variants form two clusters when superimposed on the conformation of the conserved regions. To determine if antibodies with the potential to enhance or neutralize HIV-1 SF2 change over time in infected individuals sera from chimpanzees were used because no material was still available from the original patient. Antibodies to HIV-1 SF2 neutralizing epitopes and HIV-1 SF13 enhancing epitopes were present in the circulation of chimpanzees infected with HIV-1 SF2. Once antibodies to the neutralizing epitopes were induced they persisted whereas antibodies to the enhancing epitopes varied with time after infection. Conditions may therefore exist within individual hosts where not only neutralizing but also enhancing antibodies have the potential to contribute to the selection pressure operating on the circulating population of polymorphic variants. PMID- 11424107 TI - Monitoring the response to antiretroviral therapy in HIV-1 group O infected patients using two new RT-PCR assays. AB - Failure to recognise infection caused by human immunodeficiency virus type 1 (HIV 1) group O variants has been described using both serological and genetic procedures. Moreover, monitoring the response to antiretroviral therapy is a difficult task in patients infected with HIV-1 group O since commercial tests are not available so far for the quantitation of this virus. In this study, the virological response to antiretroviral therapy were assessed in five HIV-1 group O-infected patients living in Spain by using two new and different RT-PCR methods (MUPROVAMA and LCx). Twenty-four plasma samples belonging to these five patients were selected. As reference, p24 antigenaemia levels and CD4+ cell counts were used. All samples yielded positive viral load values using MUPROVAMA (range: 138 to 595,500 HIV-RNA copies/ml) and 23 of 24 using LCx (range: < 178 to 98,356 HIV RNA copies/ml). Overall, the results obtained using both assays showed a good correlation among themselves, and in respect to p24 antigenaemia and CD4+ cell counts. However, the values provided by LCx were significantly lower (0.33 logs on average) than those provided by MUPROVAMA. In conclusion, both the highly sensitive MUPROVAMA and LCx Quantitative assays might represent an useful tool for guiding the decision on when start treatment and for monitoring the response to antiretroviral therapy in HIV-1 group O-infected patients. PMID- 11424108 TI - Phenotypic analysis of the sensitivity of HIV-1 to inhibitors of the reverse transcriptase, protease, and integrase using a self-inactivating virus vector system. AB - Conventional phenotypic analysis of resistance of the human immunodeficiency virus (HIV) to antiviral therapy is time-consuming and requires culture of infectious virus. Although phenotypic analyses may be desirable, rapid generation of test results and decentralized availability of the test system will be important to achieve utility in the clinical practice. This study describes the design of an alternative phenotypic resistance test using replication incompetent viral vectors. Chimeric HIV vectors containing a marker gene were generated. The env and most of the regulatory and accessory genes of HIV were removed. In addition, the 3'U3 region was deleted to obtain a self-inactivating construct. Cotransfection of the plasmid with a plasmid that provided the vesicular stomatitis virus glycoprotein resulted in the production of replication incompetent virus vectors. Infection of susceptible cells with the vectors led to marker gene expression. Vector production in the presence of protease (PR) inhibitors, or infection in the presence of reverse transcriptase (RT) or integrase (IN) inhibitors reduced marker gene expression in a dose-dependent manner. Marker gene activity was preserved at higher drug levels if vectors contained RT and PR genes from resistant virus isolates. Sensitivity to nucleoside and non-nucleoside RT inhibitors, protease and integrase inhibitors could be determined in 10 working days. The phenotypic drug resistance test using replication-incompetent HIV vectors significantly speeds up drug resistance measurements and allows testing at reduced biosafety levels. This will make clinical use of phenotypic assessment of antiviral resistance more feasible. PMID- 11424109 TI - Human immunodeficiency virus-1 specific and natural cellular immunity in HIV seronegative subjects with multiple sexual exposures to virus. AB - The probability of HIV infection by sexual contact, although it varies greatly, appears to be lower than that of infection by other routes of exposure. The aim of this study was to evaluate immunological determinants involved in protection against HIV infection in subjects with multiple and repeated sexual exposures to the virus. Twenty-two subjects were studied for CD8+ cell anti-HIV suppression activity and serum neutralizing activity against the HIV strain of their own partners, beta-chemokine production, and natural killer cell activity. CD8+ cell anti-HIV activity and neutralizing activity of sera were found in 13 (76%) and 12 (70.5%) out of 17 HIV-1 negative subjects, respectively. Six individuals had a relevant immune response against HIV: three subjects with a high CD8+ cell antiviral suppression activity and three individuals with sera neutralizing activity titer >1:10. These last three subjects had the highest beta-chemokine levels, a very prolonged period of multiple sexual intercourse (>6 years) and a seropositive partner with a high viral load. A partial reduction of neutralizing activity titer was observed when pre-incubating the sera with anti-beta-chemokine neutralizing antibodies. A spontaneous natural killer cell activity was suppressed in the majority of HIV-1 negative subjects with sexual exposure in comparison with normal individuals. The protection from sexual HIV transmission appears to be the result of a network of different humoral and cellular factors. PMID- 11424110 TI - 9-Nitrocamptothecin inhibits HIV-1 replication in human peripheral blood lymphocytes: a potential alternative for HIV-infection/AIDS therapy. AB - The ability of the anti-cancer drug, 9-Nitrocamptothecin (9NC), to inhibit replication of HIV-1 in clinically relevant primary lymphocytic cells was studied. Primary peripheral blood lymphocytes (PBLs) from a non-infected donor were freshly infected with HIV-1 and treated with 9NC by using three different treatment schedules. Cells were monitored for cytotoxicity by the XTT metabolic cell proliferation assay and a sensitive flow cytometric assay that was capable of measuring cell cycle changes and apoptosis. 9NC inhibited replication of HIV-1 in PBLs by greater than 95% in a dose-dependent manner as measured by the level of extracellular HIV-1 p24 release. Similar results were observed, whether 9NC was applied in a single, double, or triple dose regimen. Minimal cytotoxicity was observed for both non-infected and infected PBLs, as determined by the XTT assay. Moreover, 9NC induced apoptosis within 24 hours of drug treatment in freshly infected, but not non-infected, PBLs. The data showed that 9NC reduced replication of HIV-1 in primary human lymphocytes; thus, it indicates the potential clinical utility of this drug as an alternative or adjunct therapy for HIV-infection/AIDS. PMID- 11424111 TI - Genetic analysis of the capsid region of astroviruses. AB - Eight serotypes of human astroviruses (HAstV-1 to HAstV-8) have been described. To date, the entire genomes of HAstV-1 and HAstV-2 as well as the ORF2 sequences of HAstV-1-6 and 8 have been reported. In this study, the ORF2 sequences of seventeen strains of HAstVs originating from different countries were determined, as well as the sequence ORF2 of one porcine astrovirus (PAstV) strain. Afterwards, comparison of the capsid protein precursors encoded by ORF2 of 46 strains of HAstVs, PAstV, and feline astrovirus (FAstV) was carried out. A phylogenetic tree showed eight genogroups of HAstVs that corresponded exactly to the serotypes. HAstV-3 and 7 were the most closely related, whereas HAstVs, FAstV, and PAstV segregated from each other. Compared to a PAstV, a FAstV is closer to HAstVs. Furthermore, the capsid protein precursors were divided into four regions (after amino acid residues 424, 688, and 776, respectively) based on sequence identity. Region I was the most conserved, and FAstV was very close in identity to HAstVs. Two amino acid motifs in region I were predicted to contain the common antigenic epitopes. Region II was relatively variable. Deletions and insertions were characteristic of region III, and region IV was relatively conserved. To our knowledge, this is the first comparative sequence analysis of the capsid protein precursors of eight serotypes of HAstVs as well as two animal astroviruses (FAstV and PAstV). PMID- 11424112 TI - Characterisation of a South African human astrovirus as type 8 by antigenic and genetic analyses. AB - Human astroviruses (HAstV) can, on the basis of immunoassays using type-specific rabbit antisera, be classified into eight serotypes that correlate with genotypes. Very few isolates of HAstV type 8 have been described and there is a paucity of data available with regard to the antigenic and genetic relationships between HAstV type 8 (HAstV-8) and HAstV types 1 (HAstV-1) to 7 (HAstV-7). A wild type HAstV from a South African paediatric patient with diarrhoea was analysed antigenically, by immune electron microscopy and enzyme immunoassay, and genetically in selected regions of the ORF1a, ORF1b and ORF2 and characterised as a HAstV-8. This HAstV-8 strain exhibited greatest homology with HAstV-4 in the 5' end of the capsid gene and ORF1a and 1b, and greatest homology with HAstV-5 in the 3' end of the capsid region. This study confirms, by both antigenic and genetic analyses, that HAstV-8 represents a distinct antigenic and genotype and is the first report of a HAstV-8 from a hospitalised paediatric patient with diarrhoea in southern Africa. PMID- 11424113 TI - Symptom pathogenesis during acute influenza: interleukin-6 and other cytokine responses. AB - In experimental human influenza infection initiated by nasal inoculation, the magnitude of viral replication, fever, and symptoms correlate with nasopharyngeal lavage fluid levels of various cytokines. Our aim was to assess these relationships in patients with naturally occurring acute influenza. Patients with culture-positive influenza illness of less than 36 hr of duration were studied. Nasopharyngeal washing were collected at enrollment and on Day 2, 4, 6 and 8 for quantitative virus isolation and IL-6, TNF-alpha, INF-alpha, INF-gamma and IL-10 determinations. Blood samples collected at entry and on Day 2 and 6 were processed to assess plasma cytokines and circulating influenza RNA. Patients received either oseltamivir or placebo for 5 days. We assessed the correlation between nasopharyngeal lavage fluid or blood levels of cytokines before treatment and viral titers, symptom severity and fever. Sixteen adult subjects (median age of 22 years) were studied. In this small group of patients no significant differences between placebo and oseltamivir patients were found in viral replication or measures of cytokines. Thus the data for all 16 subjects were pooled for analysis. At entry, influenza A viruses were cultured from nasopharyngeal washes at a median titer of 4.8 log(10)TCID(50)/ml of wash. Viral titers correlated positively with symptom score (P = 0.006) and temperature values (P < 0.001). Viral titers, fever and symptoms were highest at enrollment and fell in parallel during the subsequent days. RT-PCR assays failed to detect influenza RNA in the white blood cells from any patient. We observed a significant release, in both nasopharyngeal lavage fluid and in plasma, of IL-6, TNF-alpha, INF-alpha, INF-gamma and IL-10. At entry high IL-6 levels were detected in the nasopharyngeal lavage fluid (median 10.3 pg/ml) and plasma (median 5.1 pg/ml) of all patients. We found a positive correlation between plasma IL-6 levels and both symptom scores and temperature values (P < 0.05), as well as a positive correlation between nasopharyngeal lavage fluid levels of IL-6 and TNF-alpha and temperature (P < 0.05). We did not find significant associations between symptoms, fever and levels of INF-alpha, INF-gamma or IL-10. The magnitude of early decrease in viral titers correlated with initial levels of INF-gamma in nasopharyngeal lavage fluid (P < 0.05). Significant production of IL 6, TNF-alpha, INF-alpha, INF-gamma and IL-10 occurs in response to community acquired influenza A illness. As in experimental influenza, symptoms and fever in natural acute influenza correlate with the release of IL-6. PMID- 11424114 TI - Genetic analysis of recent Taiwanese isolates of a variant of coxsackievirus A24. AB - Epidemics of acute hemorrhagic conjunctivitis (AHC) caused by a variant of coxsackievirus A24 (CA24v) reappeared in Taiwan in 1990 and 1994, following the first two epidemics of 1985--86 and 1988--89. To analyze the genetic diversity of recent CA24v in Taiwan, 7 Taiwanese strains isolated during the 1990--94 period were studied together with one Japanese and two Thai strains isolated in 1993. A fragment of 674 nucleotides between the carboxy terminal 3A and the amino terminal 3D polymerase, including the entire 3C protease (3C(pro)), was amplified by a reverse transcription-polymerase chain reaction (RT-PCR) and the nucleotide sequences were determined. In the 549 nucleotides (183 amino acids) of the entire 3C(pro), we found nucleotide differences at 80 positions between 10 strains and the prototype strain, EH24/70, one of the earliest strains of CA24v. Most of the nucleotide changes were synonymous substitutions and only nine amino acid changes were found. The nucleotide sequence homologies among 71 strains worldwide were 88 100%. These 71 nucleotide sequences were then analyzed by Neighbor-joining method and phylogenetically separated into three distinct genotypes. Genotype I consisted of early strains isolated in 1970--71 from Singapore and Hong Kong. Genotype II included isolates from Singapore and Thailand obtained in 1975. Genotype III comprised strains from the eastern hemisphere isolated in 1985--94 from Japan, Taiwan, China, Hong Kong, Thailand, Singapore, Pakistan and Ghana. They were further divided chronologically into six clusters. The recent isolates from Taiwan obtained in 1985/1986, 1988/1989 and 1990--94 were classified into genotype III Clusters 1, 5, and 6 respectively. The evolutionary rate was re estimated to be 3 x 10(- 3) 30 years after the emergence of the virus. PMID- 11424115 TI - Differences between the quantitative antigenemia assay and the cobas amplicor monitor quantitative PCR assay for detecting CMV viraemia in bone marrow and solid organ transplant patients. AB - The relationship between quantitative PCR (COBAS Amplicor CMV Monitor, Roche Diagnostics) and quantitative antigenemia (Monofluor pp65, Sanofi Diagnostics) was examined for monitoring CMV viraemia. A total of 469 specimens from immunocompromised haematology and solid organ transplant patients were tested by quantitative antigenemia and qualitative PCR. Quantitative PCR (QPCR) was performed on the 245 specimens in which CMV DNA was detected by qualitative PCR. To exclude any effect due to specific anti-CMV treatment, analysis of antigenemia and QPCR results was only performed on the 164 of 245 specimens collected from patients not on ganciclovir or foscarnet treatment. Forty seven specimens had <400 CMV copies/mL and a negative antigen result, four specimens were antigen positive (all between 1 to 10 positive CMV cells/2 x 10(5) leucocytes) and had <400 CMV copies/mL. Fifty-one specimens had a CMV viral load > or = 400 copies/mL and a negative antigen result and 62 specimens had a CMV viral load > or = 400 copies/mL and a positive antigen. The viral load was shown to be as high as 43,000 copies/mL in some patients with a negative antigen and occurred in non neutropenic patients. The correlation coefficient for antigen and QPCR results for specimens from bone marrow transplant patients, was 0.69 with an average CMV viral load of 3,200 copies/mL (SEM = 800) and an average antigen of nine positive CMV cells/2 x 10(5) leucocytes (SEM = 3). In the corresponding solid organ transplant group, the correlation coefficient for antigen and QPCR results was 0.71 with an average CMV viral load of 9,900 copies/mL (SEM = 2,100) and an average antigen of 26 positive CMV cells/2 x 10(5) leucocytes (SEM = 6). Both the average viral load and the average antigen result in specimens from solid organ transplant patients, were significantly higher than the average viral load and antigen result in the corresponding group of bone marrow transplant patients (Two Sample-for-Means z-Test, P = 0.001 and P = 0.003, respectively). The differences in the kinetics of the two assays in monitoring CMV and their ability to predict CMV disease was also assessed in a sub-group of patients. In conclusion, the two assays used in this study do not always show parallel changes in CMV viral load, but may be complementary for the diagnosis and management of CMV disease. The observation that non-neutropenic patients can have a high viral load in plasma and a negative antigenemia has implications for laboratories using antigenemia alone to monitor patients for CMV disease. PMID- 11424116 TI - Quantity of cytomegalovirus DNA in different leukocyte populations during active infection in vivo and the presence of gB and UL18 transcripts. AB - The quantity of human cytomegalovirus (HCMV) DNA in the blood of immunocompromised individuals correlates with the development of HCMV disease. We wished to determine which leukocytes harboured DNA and whether this represented active viral replication. Magnetic bead separation techniques were used to obtain pure polymorphonuclear leukocyte (PMNL), monocyte, B and T cell fractions, and RT PCR and quantitative-competitive PCR (QC-PCR) to detect HCMV glycoprotein B (gB; UL55) transcripts and quantify HCMV DNA levels, respectively, in each cell fraction. QC-PCR revealed that PMNLs contribute the greatest to the overall viral load in blood (median viral load: PMNLs, 10(5.37) genomes/ml of blood; monocytes, 10(4.40) genomes/ml; B cells, 10(3.70) genomes/ml; and T cells, 10(4.08) genomes/ml). However, monocytes have a viral burden of 0.65 genomes/monocyte which is greater than that within the other leukocyte populations (0.11 genomes/PMNL, 0.23 genomes/B cell, and 0.20 genomes/T cell). Glycoprotein B transcripts were detected in all four cell populations: 3/10 PMNL fractions, 6/13 monocyte fractions, 5/13 B cell fractions, and 4/13 T cell fractions. The data show that productive infection of these leukocyte subpopulations, including PMNLs, can occur in vivo. Furthermore, transcripts of gpUL18, the putative natural killer (NK) cell decoy, were detected in 2/6 monocyte fractions with active replication, and 1/4 T cell fractions but not in the other leukocyte fractions. The transient nature of UL18 gene expression, and the low abundance of the transcript relative to gB were confirmed. PMID- 11424117 TI - Comparison of a triple antigen and a single antigen recombinant vaccine for adult hepatitis B vaccination. AB - Hepatitis B and its sequelae are a major public health problem. Vaccines have been available for almost 20 years; however the disease still remains a global problem. Many factors contribute to the failure to control hepatitis B, including the limited nature of the vaccination programs implemented initially. Only relatively recently has mass childhood vaccination begun to be implemented and vaccination of high-risk groups, other than healthcare workers, is still not general policy. Additional factors contributing to continued persistence of hepatitis B in the developed world are that the present vaccines are not fully used by those recommended to be vaccinated and even when vaccination is carried out appropriately, there remain some who fail to achieve adequate protection. Clearly, the protection of at-risk groups who have inadequate response to current vaccines, and those who are unwilling or unable to comply with protracted multi dose vaccine regimens, could be improved if there were a more potent vaccine and/or a shorter vaccination regimen available. Adults who had never been vaccinated against hepatitis B were randomised to receive a vaccination course of either a present single antigen (S) vaccine (Recombivax-HB) or a novel triple antigen (S, pre-S1, and pre-S2) recombinant vaccine (Hepacare Medeva Pharma plc). Doses were given at baseline and 1 month and 6 months later. Hepatitis B surface antibody (anti-HBs) levels were measured at 2, 4, 6, and 7 months after beginning vaccination. The primary efficacy parameter was the degree of protection, measured as the percentage of subjects with anti-HBs titres > or = 10 IU/L, 6 or 7 months (26 +/- 2 weeks) after beginning vaccination. A total of 303 adult subjects entered the study and were vaccinated. Of these, 11 failed to complete the study (4 on Hepacare and 7 on Recombivax-HB); however all but 2 (1 to receive the triple antigen vaccine and 1 to receive Recombivax-HB) were included in the intent-to-treat population for efficacy evaluation. Treatment randomisation was stratified at entry based on age (above and below 40 years old) and gender. The standard three-dose/6-month vaccination regimen of the single antigen vaccine protected 83% of subjects by 7 months after starting vaccination whereas the triple antigen vaccine as a two-dose/1-month regimen protected 88% within 6 months and as a three-dose/6-month regimen protected 97% by 7 months after starting vaccination. Thus the protection rate provided by the shortened (0, 1) regimen of the novel vaccine was "essentially equivalent" (i.e., not statistically inferior) to that provided by the full (0, 1, and 6) regimen of today's vaccine (88% vs. 81%, P < 0.001), and the protection rate provided by a three-dose/6-month (0, 1, and 6) regimen of the new vaccine was significantly superior to that provided by present vaccines (97% vs. 83% P < 0.001). The percentage of subjects protected increases with time after beginning vaccination and at all time points up to and including 6 months was significantly greater with the two-dose regimen of the triple antigen vaccine than with the single antigen vaccine regimen. In adults at risk for a reduced response to hepatitis B vaccination [i.e., older adults (>/=40), the obese, males, and smokers], the triple antigen vaccine produced a significantly greater percentage of protected subjects (P < 0.001) and higher geometric mean titre (P < 0.001). Indeed as a three-dose/6 month regimen, the triple antigen vaccine raised the level of protection in these vulnerable subgroups to that seen when a single antigen vaccine is used in the optimal younger adult group. Both vaccines were well tolerated and had similar safety profiles. The most frequently (> or = 10%) reported adverse events with the use of either vaccine were pain at the site of injection (38% vs. 41% vs. 20% for the two-dose Hepacare regimen, the three-dose Hepacare regimen, and the three-dose Recombivax-HB regimen, respectively), infections at the site of injection (1% vs. 14% vs. 9%), headache (9% vs. 13% vs. 11%), and nausea (7% vs. 11% vs. 3%). It is concluded that in healthy normal adults, a triple antigen hepatitis B vaccine that contained S and pre-S antigens produced an enhanced immunological response. This was exemplified by the novel vaccine's ability to overcome factors such as advancing age (> or = 40 years), obesity, and smoking, each of which is known to reduce the potential for protection with present recombinant S only vaccines. A two-dose/1-month (0 and 1) regimen of this triple antigen vaccine was as effective as the standard three dose/6 month (0, 1, and 6) regimen of present single antigen vaccines. (c) 2001 Wiley-Liss, Inc. PMID- 11424118 TI - Full-length genomic analysis of hepatitis B virus isolates in a patient progressing from hepatitis to hepatocellular carcinoma. AB - In hepatitis B virus (HBV)-endemic countries, the majority of hepatocellular carcinoma (HCC) arises in HBV carriers. High frequency of mutations at nucleotides 1762(A-->T) and 1764(G-->A) in the core promoter region have been described in HCC. Due to the differences in genetic backgrounds, environmental risk factors and random cellular insertion sites, it is difficult to analyze the possible roles of HBV variants detected in different HCC patients. In a follow-up cohort study, an HBsAg-positive asymptomatic carrier was diagnosed HCC within 4 years. Eleven full-length HBV isolates, three from the first serum sample obtained 4 years pre-HCC, and eight from serum sample, peri-tumor and tumor tissue post-HCC of this individual were sequenced and used to transfect HepG2 cells. When sequences were compared between pre- and post-HCC isolates, no single mutation common to all post-HCC isolates that differed from pre-HCC isolates was found. Among all 11 isolates, there were 20 predicted amino acid substitutions shared by two or more post-HCC isolates. These were located in the S(5), X(4), core(4), polymerase(4), pre-S1(2) and pre-S2(1) proteins. Possible roles of amino acid substitutions and enhanced replication efficiency in cells transfected by post-HCC isolates are discussed. PMID- 11424119 TI - Hepatitis B virus genotypes and HBsAg subtypes in refugees and injection drug users in the United States determined by LiPA and monoclonal EIA. AB - Hepatitis B virus (HBV) genotyping and hepatitis B surface antigen (HBsAg) subtyping were carried out on sera from 196 HBsAg-positive patients, including 151 refugees entering the United States and 45 injection drug users in Seattle. HBsAg subtyping was performed by enzyme immunoassay (EIA) using a panel of monoclonal antibodies and the HBV genotype was determined by polymerase chain reaction (PCR) followed by detection of amplified HBV DNA by a reverse-phase hybridization line probe assay (LiPA) using genotype-specific probes. HBV DNA was detected by PCR in 155 (79%) of the 196 sera and all 155 were genotyped by LiPA. Samples from Southeast Asia were predominantly genotype B/subtype ayw1 and genotype C/adr; samples from the former Soviet Union and eastern Europe were mostly genotype D/ayw2 and genotype D/ayw3; samples from east Africa were mainly genotype A/adw2 and genotype D/ayw2; and samples from injection drug users were mostly genotype D/ayw3 and genotype A/adw2. Some strains of ayw3 gave atypical monoclonal antibody reactivity patterns in the subtyping assay due to a Val/Ala instead of a Thr at amino acid residue 118 and a Thr instead of a Met at residue 125. A strain of ayw2 also gave an atypical monoclonal antibody reactivity pattern due to an Ala instead of a Thr at amino acid residue 123. LiPA genotyping and monoclonal EIA subtyping can provide useful information for epidemiological studies. PMID- 11424120 TI - Hepatitis B virus markers in anti-HBc only positive individuals. AB - Isolated reactivity to hepatitis B virus (HBV) core antigen (anti-HBc) is observed relatively frequently in immunocompromised individuals, intravenous drug abusers (IVDA), and in the presence of HCV infection. The reason for the lack of HBsAg is not clear. The aim of the present study was to investigate which factors (genetic variability of S gene, low-level HBsAg, and immune complexes may be responsible for the failure of HBsAg detection with commercial HBsAg screening assays. Dilution series of two recombinant HBsAg escape mutants and dilutions of serum samples from chronic HBV carriers with multiple insertions in the a determinant and different HBsAg subtypes were tested with a highly sensitive assay that detects wild-type HBsAg (Elecsys HBsAg, Roche Diagnostics, Penzberg, Germany) and two assays that detect HBV wild-type and escape mutants (Murex HBsAg Version 3, Murex and Enzygnost HBsAg 5.0, Dade Behring, Marburg, Germany). Elecsys HBsAg showed in comparison to Murex HBsAg Version 3 and Enzygnost HBsAg 5.0 a reduced sensitivity for escape mutant detection. On the other hand, the best performance for HBsAg subtype detection was obtained with Elecsys HBsAg. In the second part of the study, a selected panel of isolated anti-HBc reactive (n = 104) serum samples (AxSYM Core) was submitted to testing by Elecsys HBsAg, Murex HBsAg Version 3, Enzygnost HBsAg 5.0, and HBsAg detection after immune complex dissociation (ICD) and anti-HBs determination with two different assays (AxSYM Ausab and Elecsys Anti-HBs). To assess the specificity of anti-HBc test results, all the samples were tested by a second anti-HBc assay (Elecsys Anti-HBc). Quantitative HBV DNA detection was undertaken with a commercially available HBV PCR assay (Amplicor HBV Monitor). HCV infection was present in 65.4% of anti-HBc only reactive individuals. Five AxSYM Core positive samples were negative by Elecsys Anti-HBc. Overall, 15 (14.4%) AxSYM Ausab negative samples gave positive results with Elecsys Anti-HBs (median value: 21 IU/ml). No low-level HBsAg carrier was detected among the isolated anti-HBc reactive individuals with Elecsys HBsAg. There was no evidence for the presence of immune complexes. Only one sample was repeatedly reactive by the Murex HBsAg, suggesting that the a mutant form of HBsAg was responsible for the isolated anti-HBc reactivity, however neutralisation assay was not interpretable and HBV DNA PCR was negative. Fifteen (14.4%) anti-HBc only positive individuals were HBV DNA carriers with concentrations ranging from 800 to more than >4,000,000 copies of viral DNA/ml. In conclusion, the most probable explanations for isolated anti-HBc reactivity in our study group are a possible interference of HBsAg synthesis by HCV infection (65.4%) and divergence of results of anti-HBs assays (14.4%). There is no evidence for the presence of low-level HBsAg carriers and immune complexes. HBsAg mutants cannot be excluded definitively by the test strategy used in the present evaluation. PMID- 11424121 TI - Strategies for reliable diagnosis of hepatitis C infection: the need for a serological confirmatory assay. AB - The aim of the study was to examine whether the diagnosis of Hepatitis C (HCV) infection can be obtained reliably without using an immunoblot-based confirmation assay. 1,708 EIA-reactive serum samples were examined retrospectively for (i) optical density value in the screening assay, (ii) reactivity in an immunoblot assay, and (iii) result by RT PCR. In 1,394 (81.0%) samples positive results were obtained by both the HCV EIA and the confirmation assay. OD-values > or = 2.2 were observed in 1026 of these samples, but covered the range from 0.4 to 2.1 in the other 368 samples. The combination of HCV EIA reactivity and indeterminate immunoblot assay was observed in 134 (7.8%) serum samples. HCV RNA was detected in 58 cases by PCR. The OD-values of these 58 samples ranged from 0.4 to >2.2. Especially reactivity against the core recombinant protein was indicative of PCR positivity. The reactivity by the HCV EIA could not be confirmed by immunoblot assay or PCR in 180 (10.5%) sera. These false reactive sera showed OD values by EIA from 0.3 to 2.1. It is concluded that no threshold values can be defined which would allow differentiation between positive, indeterminate, and false reactive result by HCV EIA without producing an unacceptably high number of false negative diagnoses. Not using immunoblot-based confirmation would result in many additional PCR examinations. Therefore, confirmation of reactive HCV EIA results by a serological confirmatory assay must remain an essential part of the diagnostic procedure. PMID- 11424122 TI - Genetic characterization of hypervariable region 1 in patients chronically infected with hepatitis C virus genotype 2. AB - The hypervariable region 1 (HVR1) has been most reliably identified in the genome of HCV genotype 1 isolates and thought to possibly play a role in immune evasion and development of chronic infection. There are few studies, however, of other HCV genotypes to determine if they also have such a hypervariable region present, and it is unclear whether or not there is any genotype-dependent difference in the genetic characteristics of HVR1. We determined the nucleotide sequence of 5' end of E2/NS1 region of the HCV genome spanning HVR1 of multiple genotype 1 and 2 HCV isolates and carried out a detailed genetic analysis. Similarity plots identified two hypervariable regions within the genotype 2 sequences, a larger one corresponding to HVR1 as well as a smaller 27-nucleotide region of hypervariability. The synonymous substitutions per synonymous site (ds) was greater than nonsynonymous substitutions per nonsynonymous site (dn) within genotype 1 group whereas dn and ds were similar in the genotype 2 group. Analysis of amino acid sequences revealed several conserved sites across genotype 1 and 2 (amino acid numbers 2,6, 20 and 26) and overall similar hydropathic profiles were found within two genotypes. Still, despite the hypervariability, the HVR1 showed a genotype-specific phylogenetic clustering. Thus, HVR1 appears to be conserved between genotypes in keeping with it having an important survival function. Genotype 2 appears to have a greater rate of nonsynonymous substitutions within HVR1, suggesting a greater positive evolutionary pressure. PMID- 11424123 TI - Sequence analysis of hepatitis C virus isolated from a fulminant hepatitis patient. AB - Although hepatitis C virus (HCV) is a major cause of non-A non-B hepatitis, its pathogenic role in fulminant hepatitis remains controversial. A 32-year-old man contracted hepatitis. Serum ALT concentration was reached to 6,970 IU/L, the lowest prothrombin time value was 16% and jaundice and stage II encephalopathy were developed. HCV RNA was detected in this patient by reverse transcription polymerase chain reaction in sera at the acute phase, and it was undetectable during the remission phase when anti-HCV was found. The entire genome of infected HCV was recovered, cloned, and sequenced from this patient, and compared with the clones of six other chronic hepatitis patients. Phylogenetic analysis revealed a clustering around genotype 2a and a deviation from the other 2a chronic hepatitis strains. Calculating the genetic distance in each subgenomic region revealed that the 5'untranslated region (5'UTR), core, nonstructural (NS) 3, and NS5A were severely deviated. Of 20 clones of the hypervariable region (HVR), 17 showed an identical sequence with the others showing a difference of only one amino acid. HCV was isolated from a fulminant hepatitis patient and its entire genome was recovered; a clustering around genotype 2a was observed, but the sequence deviated especially in 5'UTR, core, NS3, and NS5A; and monoclonality of the HVR sequence was found not only in the fulminant hepatitis patient but in a certain percentage of chronic hepatitis patients. PMID- 11424124 TI - Quantitative analysis of specific Th1/Th2 helper cell responses and IgG subtype antibodies in interferon-alpha-treated patients with chronic hepatitis C. AB - This study aimed to characterise the immune mechanisms relevant to viral clearance in interferon (IFN)-alpha-treated chronic hepatitis C virus (HCV) infection. Proliferative responses of peripheral blood mononuclear cells from sustained complete IFN-alpha therapy responders (n = 8), nonresponders (n = 13), untreated patients (n = 10), and healthy controls (n = 5) were measured retrospectively upon stimulation with recombinant HCV-antigens (core, helicase, NS3, NS4, and NS5) and the secretion of IFN-gamma and interleukins (IL-4, IL-5, IL-10, and IL-12) were tested by ELISA. Furthermore, IFN-gamma as well as IL-10 secreting CD4+ T cells were quantitated by intracellular cytokine staining. Anti HCV core and NS3-specific IgG subclass antibodies were quantitated in the corresponding patient sera. Sustained therapy responders had more frequent and stronger NS3 and helicase-specific cellular immune responses than nonresponders, untreated HCV patients and healthy controls. Independent from therapy outcome HCV stimulated T cells in IFN-alpha treated patients secreted preferentially IFN gamma The Th2 cytokines IL-4 and IL-10 were even decreased in nonresponders, while the IL-12 secretion was not influenced. With respect to the humoral immune response sustained complete responders showed significantly reduced IFN-gamma independent anti-HCV-core and -NS3 IgG1 antibody synthesis. In conclusion, vigorous NS3-specific T-helper cell responses were associated with viral clearance in IFN-alpha recipients; however, the cytokine and antibody analysis argues against a Th1/Th2 imbalance as a major factor that influence the therapy outcome. PMID- 11424125 TI - HCV genotype and "silent" HBV coinfection: two main risk factors for a more severe liver disease. AB - To evaluate whether HCV genotype and a "silent" HBV infection may be related to a more severe clinical presentation of liver disease, 205 anti-HCV/HCV-RNA positive, HBsAg/anti-HBs negative patients with chronic hepatitis (113 males and 92 females; median age 55 years, range 18-77), were studied on presentation at the Liver Unit from January 1993 to December 1997. Presence of serum anti-HBc, in the absence of HBsAg and anti-HBs, was considered a marker of "silent" HBV infection. Of the 205 patients, 134 had undergone percutaneous liver biopsy. Two main diagnosis groups were established: the mild liver disease group (76 patients), and the severe liver disease group (109 patients); 20 patients who had refused to undergo liver biopsy were not included in the clinical and virological evaluation because the diagnosis was uncertain. The prevalence of severe liver disease was similar in the genotype 1 and non-1 groups (61.3% of 98 patients with genotype 1 and 52.9% of 70 patients with a non-1 genotype). Instead, the 88 patients with "silent" HBV infection showed a higher percentage of severe liver disease than the 97 anti-HBc negative patients (72.7% vs. 46.4%, respectively: P < 0.0005). Of the 88 anti-HBc positive patients, the prevalence of those with severe liver disease was similar in the 32 cases with serum HBV-DNA as detected by PCR and in the 56 HBV-DNA negative (81.2% vs. 67.8%, P = 0.4). The relation between "silent" HBV infection and severe liver disease was observed both in genotype 1 and non-1 infected patients. Nevertheless, the anti-HBc negative patients infected by genotype 1 showed a severe liver disease more frequently than those infected by a non-1 genotype, with a difference that is significant to the statistical analysis (P < 0.05). The findings suggest that "silent" HBV infection in anti-HCV positive chronic hepatitis enhances the severity of the liver disease. Evidence was also found that in patients without "silent" HBV infection there is a correlation between the presence of HCV genotype 1 and the severity of liver disease. PMID- 11424126 TI - Hepatitis delta virus genotypes I and III circulate associated with hepatitis B virus genotype F In Venezuela. AB - The genotypes of hepatitis B (HBV) and delta (HDV) viruses circulating among Venezuelan Amerindian populations, where these viruses are endemic, were determined by sequencing of PCR amplified products from HBsAg positive sera. HDV genotype I (n = 7, 6 from West Amerindians), and III (n = 5, 4 from South Amerindians), were found. Only one HDV genotype I isolate was associated with HBV genotype D, 4 HDV genotype I and 2 HDV genotype III infected individuals were co infected with HBV genotype F. The failure to detect the South American HDV genotype III in West Amerindians might be related to the outbreak of fulminant hepatitis with high mortality rate occurred between 1979 and 1982, probably affecting more the Amerindians infected with HDV genotype III. These results suggest the circulation of HDV genotype I among Amerindians, probably introduced through European immigrations, and that this HDV genotype is able to replicate in association with HBV genotype F. PMID- 11424127 TI - Diagnosis of human parvovirus B19 infections by detection of epitope-type specific VP2 IgG. AB - In the B19 VP2 molecule an immunodominant heptapeptide epitope has been detected, recently for which IgG antibodies are synthesized exclusively in the acute phase of B19 infection. Using this acute-phase-specific epitope (KYVTGIN) a 2(nd) generation epitope-type EIA was developed, which compares serum IgG activity for native VP2 capsids exhibiting conformational VP2 epitopes with IgG activity for the KYVTGIN epitope. In this study the diagnostic performance (clinical sensitivity and specificity) of the 1st and 2nd-generation epitope-type EIAs and of a peptide-based EIA utilising as antigen the KYVTGIN epitope alone was assessed in comparison with various high-quality IgM- and IgG- based B19 assays. Serum samples from 489 patients with B19-related symptoms and asymptomatic controls from three countries were studied. Among 323 patients with B19-IgG, 20% were diagnosed as acute infection, 73% had past immunity and 7% were not classified due to contradictory results among the different assays. The unclassified samples were explored for viral strain diversity by PCR and DNA sequencing but all sequences obtained were B19-like with variance of only a few per cent. The 2nd-generation epitope-type EIA had a diagnostic sensitivity of 98% and a diagnostic specificity of 94%. In combination with conventional approaches, the epitope-type assays increase greatly the accuracy of B19 serodiagnosis. PMID- 11424128 TI - Identification of a DNA encapsidation sequence for human polyomavirus pseudovirion formation. AB - Human polyomavirus is a naked capsid virus containing a closed circular double stranded DNA genome. The mechanism of DNA encapsidation for the viral progeny formation is not fully understood. In this study, DNA encapsidation domain of the major capsid protein, VP1, of the human polyomavirus JCV was investigated. When the first 12 amino acids were deleted, the E. coli expressed VP1 (Delta N12VP1) failed to encapsidate the host DNA although the integrity of the capsid-like structure was maintained. In addition, capsid-like particles of Delta N12VP1 did not package exogenous DNA in vitro, which is in contrast to that of the full length VP1 protein. These findings suggest that the N-terminal of the first 12 amino acids of VP1 were responsible for DNA encapsidation. The importance of amino acids in the DNA encapsidation domain was determined further using site directed mutagenesis. All of the positively charged amino acids at the N-terminal region of VP1 were essential for DNA encapsidation. The results indicate that the N-terminal region of the human polyomavirus major capsid protein VP1 may be involved in viral genome encapsidation during progeny maturation. PMID- 11424129 TI - Genetic variability of respiratory syncytial virus subgroup a strain in 15 successive epidemics in one city. AB - The genetic variability of 125 respiratory syncytial virus (RSV) subgroup A isolates over 15 successive epidemics from 1980 to 1995 in an urban population of Japan was determined. Allocation of isolates into lineages was archived by reverse transcriptase-polymerase chain reaction amplification of selected regions of the nucleoprotein (NP) and attachment (G) protein gene followed by restriction fragment length polymorphism (RFLP) analysis. Three and seven distinct restriction patterns of the NP and G gene were observed, respectively. When the NP and G gene RFLP analyses were combined, ten different genetic lineages were identified in the 125 isolates. The strains with the same genotype were isolated in each epidemic and the dominant lineages were replaced by others after every one to three consecutive epidemics. Nucleotide and amino acid sequencing of the variable region of G gene of these predominant isolates revealed differences of 5 -28% between strains. There was, however, no apparent accumulation of diversity with age to indicate progressive changes. The dominant strains were often closely related to those isolated in other parts of the world at a similar time. These observations suggest that dominant RSV strains are replaced frequently by others that have been co-circulating or have recently entered the community from a worldwide reservoir. The change of dominant strains may be influenced by the buildup of immunological resistance in the community to successive epidemics of the same strain. PMID- 11424130 TI - High prevalence of TT virus-related DNA (90%) and diverse viral genotypes in Norwegian blood donors. AB - Early estimates of the prevalence of TTV viremia in healthy adults of developed countries were in the order of 1--10 %, while similar estimates in Third World countries were considerably higher. Using three different PCRs, TTV-related DNA was detected in serum from 180 out of the 201 Norwegian blood donors tested, indicating that these viruses are almost universally present in adults. Sequence analysis revealed heterogeneity similar to what is found world-wide. The data suggest that the previous discrepancy in prevalences might be related to a lower serum concentration of virus in developed countries. The high prevalence adds evidence to the benign nature of the virus. PMID- 11424131 TI - Premature calvarial synostosis and epidermal hyperplasia (Beare-Stevenson syndrome-like anomalies) resulting from a P250R missense mutation in the gene encoding fibroblast growth factor receptor 3. AB - We report on a patient with a severe premature calvarial synostosis and epidermal hyperplasia. The phenotype was consistent with that of a mild presentation of Beare-Stevenson syndrome but molecular analysis of the IgIII-transmembrane linker region and the transmembrane domain of the gene encoding the FGFR2 receptor, revealed wild-type sequence only. Subsequently, molecular analysis of the FGFR3 receptor gene identified a heterozygous P250R missense mutation in both the proposita and her mildly affected father. This communication extends the clinical spectrum of the FGFR3 P250R mutation to encompass epidermal hyperplasia and documents the phenomenon of activated FGFR receptors stimulating common downstream developmental pathways, resulting in overlapping clinical outcomes. PMID- 11424132 TI - Ectodermal dysplasia, ectrodactyly and macular dystrophy (EEM syndrome) in siblings. AB - We report a brother and sister with ectodermal dysplasia, ectrodactyly, and macular dystrophy (the EEM syndrome). Both children had abnormalities of the hands and the hair, and bilateral macular degeneration. The clinical picture in both is similar to, but less severe than, that described in the previously reported cases of this rare syndrome. Even though the parents are not related, they are both of Jewish Yemenite origin, and the possibility of a common ancestor cannot be ruled out. This would suggest autosomal recessive inheritance. The clinical picture in these patients suggests either variable expression or genetic heterogeneity in the EEM syndrome and further delineates the clinical and genetic spectrum of this condition. PMID- 11424133 TI - SMN2-deletion in childhood-onset spinal muscular atrophy. AB - The human genome has two homologous survival motor neuron genes, SMN1 and SMN2. Although deletions of SMN1 are frequently reported in childhood-onset spinal muscular atrophy (SMA), SMN2 have been found to be intact in patients with the disorder. We report on a 5-year-old boy with childhood-onset SMA who has a homozygous deletion of SMN2. He had wasting, weakness, and hyporeflexia, predominantly in the distal muscles. The muscles involved showed chronic neurogenic changes on electromyogram. There was no sensory involvement. A nerve conduction study showed near normal conduction velocity with reduction in the amplitude of the compound muscle action potential. Analysis of polymerase chain reaction-restriction fragment length polymorphism as well as single-strand conformation polymorphism on exons 7 and 8 of the SMN genes revealed the SMN2 deletion. Base sequencing and densitometric analysis of the critical region (exon 7) did not show any microdeletion or duplication of SMN1, but confirmed the deletion of SMN2. We conclude that a deletion of SMN2 may also result in the SMA phenotype. PMID- 11424134 TI - Metachromatic leukodystrophy in the Navajo: fallout of the American-Indian wars of the nineteenth century. AB - Our aim was to determine if the high frequency of metachromatic leukodystrophy (MLD) in Navajo Indians of the Southwestern United States is the result of a "genetic bottleneck" that occurred in the mid 19th century. Navajo Nation, Indian Health Service, and other national databases were queried for Native American patients with MLD. Pedigrees, including birth location, were established by interviewing relatives. We found that cases of MLD and their ancestors are clustered in a portion of the western Navajo Nation to which a small number of Navajo fled after armed conflict with the United States Army in the 1860s. The observed incidence of MLD on the western Navajo Nation is 1/2,520 live births, with an estimated carrier frequency of 1/25 to 1/50. No cases were observed in the eastern part of the Navajo Nation over a period of 18 years (60,000 births). The high incidence of MLD in the western Navajo Nation appears to be the result of a genetic bottleneck and probable founder effect from the mid 19th century: This mechanism may also explain the high incidence of a number of other unique, heritable disorders among the Navajo. The history of the Navajo may also be relevant to other American Indian and Alaskan Native groups that have undergone severe population reduction since the arrival of Europeans in North America. PMID- 11424135 TI - Microphthalmia, facial anomalies, microcephaly, thumb and hallux hypoplasia, and agammaglobulinemia. AB - We report a boy, born to consanguineous patients, with agammaglobulinemia associated with multiple physical anomalies: mild intrauterine growth retardation, extreme microphthalmia (clinical anophthalmia), severe microcephaly, blepharophimosis, long face with temporal narrowing, scaphocephalic skull shape, posterior cleft palate, hypoplastic, adducted thumbs with small nails, and short, inward turned halluces with absent distal phalanges and nails. Psychomotor development was moderately delayed. No mutations were found in exons of BKT and PAX-5 genes. This unreported constellation could represent a novel, autosomal recessive syndrome. PMID- 11424136 TI - Gerodermia osteodysplastica and wrinkly skin syndrome: are they the same? AB - Gerodermia osteodysplastica (GO) is a connective tissue disorder characterized by premature aging, wrinkled, and lax skin with reduced elasticity which is more marked on the dorsum of the hands and feet associated with hyperextensible joints and osteoporosis. The wrinkly skin syndrome (WSS) is characterized by wrinkled skin over the dorsum of the hands, feet, and abdomen; hyperextensible joints, particularly of the hands; intrauterine growth retardation; postnatal failure to thrive; and mental and developmental delay. We report on five children from two consanguineous Arab families with features overlapping both GO and WSS. All five children had similar dysmorphic facial features consisting of broad and prominent forehead, hypotelorism with epicanthal folds, prominent bulbous nose, flat malar region, and large protruding ears. All had wrinkling of the skin more marked on the dorsum of the hands, feet, and abdomen; hyperextensibility of the joints, particularly of the hands; and aged appearance. Intrauterine growth retardation, subsequent failure to thrive, developmental delay, and variable degree of osteoporosis was also present in all of them. The older three children developed progressive prognathism. We suggest that GO and WSS could represent variable manifestation of the same disorder. PMID- 11424137 TI - Chest wall hamartoma with Wiedemann-Beckwith syndrome: clinical report and brief review of chromosome 11p15.5-related tumors. AB - A girl born with a left chest wall hamartoma, macroglossia, nevus flammeus of the middle forehead, and a small umbilical hernia developed left lower extremity hemihypertrophy by 1 year of age and is assumed to have Wiedemann-Beckwith syndrome. Hamartoma of the bladder and a cardiac fibrous hamartoma have been reported previously in association with Wiedemann-Beckwith syndrome. Infantile hamartomas are exceedingly rare and add to the spectrum of tumor formation in the syndrome. PMID- 11424138 TI - Clinical and molecular-cytogenetic studies in seven patients with ring chromosome 18. AB - We report the results of detailed clinical and molecular-cytogenetic studies in seven patients with ring chromosome 18. Classical cytogenetics and fluorescence in situ hybridization (FISH) analysis with the chromosome 18 painting probe identified five non-mosaic and two complex mosaic 46,XX,dup(18)(p11.2)/47,XX,dup(18)(p11.2),+r(18) and 46,XX,dup(18)(p11.32)/47,XX,dup(18)(p11.32),+r(18) cases. FISH analysis was performed for precise characterization of the chromosome 18 breakpoints using chromosome 18-specific short-arm paint, centromeric, subtelomeric, and a panel of fifteen Alu- and DOP-PCR YAC probes. The breakpoints were assessed with an average resolution of approximately 2.2 Mb. In all r(18) chromosomes, the 18q terminal deletions ranging from 18q21.2 to 18q22.3 ( approximately 35 and 9 Mb, respectively) were found, whereas only in four cases could the loss of 18p material be demonstrated. In two cases the dup(18) chromosomes were identified as inv dup(18)(qter-->p11.32::q21.3-->qter) and inv dup(18)(qter-->p11.32::p11.32- >p11.1: :q21.3-->qter)pat, with no evidence of an 18p deletion. A novel inter intrachromatid mechanism of formation of duplications and ring chromosomes is proposed. Although the effect of "ring instability syndrome" cannot be excluded, the phenotypes of our patients with characteristic features of 18q- and 18p- syndromes are compared and correlated with the analyzed genotypes. It has been observed that a short neck with absence of cardiac anomalies may be related to the deletion of the 18p material from the r(18) chromosome. PMID- 11424139 TI - Characterization of a long-term survivor with Stuve-Wiedemann syndrome and mosaicism of a supernumerary marker chromosome. AB - Stuve-Wiedemann syndrome (SWS) is typically lethal in the neonatal period; only two patients have been reported with a longer survival. We report a new patient with SWS, who at 9 years of age is one of the longest survivors with this disorder. In addition to the characteristic features of SWS, she has a number of unique clinical signs, including lack of corneal and patellar reflexes, a smooth tongue with no fungiform papillae, chronic gingival abscesses, mottled, poor dentition, blotchy pigmentation of the skin, unusual infections, multiple fractures, and progressive scoliosis. Cytogenetic analysis identified mosaicism for a supernumerary marker chromosome (SMC), seen in the majority of amniocytes, blood, and skin fibroblasts. The SMC was shown to be derived from chromosome 5 and contains euchromatin. The significance of the SMC to the etiology of SWS is unknown. This patient further demonstrates that SWS is not universally lethal. PMID- 11424140 TI - Smoking, folate and methylenetetrahydrofolate reductase status as interactive determinants of adenomatous and hyperplastic polyps of colorectum. AB - Most studies demonstrate increased risk of colorectal cancer (CRC) and adenomas in folate-deficient subjects or that high folate intake may afford some protection. Smoking increases such risk in some but not all studies. We investigated whether smoking, folate status and methylenetetrahydrofolate reductase (MTHFR) genotype predict the risk of adenomatous and hyperplastic polyps of colorectum. By colonoscopy, the type, number, size and extent of dysplasia of colorectal polyps were assessed in 443 subjects aged 63-72 years. We also determined RBC folate and the C667T polymorphism of the methylenetetrahydrofolate reductase (MTHFR) gene. Smoking, folate status and the C677T MTHFR polymorphism were strong, interactive determinants of high-risk adenomas (HRAs, defined as adenomas > or =10 mm in diameter, adenomas with villous components or with severe dysplasia). The risk was particularly high in smokers with low folate and the CT/TT genotype (risk category T) and in smokers with high folate and the CC genotype (risk category C). With non-smokers with low folate and the CC genotype as reference, the odds ratios (OR, 95% CI) were 8.7 (2.5-29.7) in category T and 9.9 (2.6-38.4) in category C. Notably, this risk pattern was also observed for hyperplastic polyps. In conclusion, in smokers, high folate status may confer increased or decreased risk for HRAs, depending on the MTHFR genotype. These data demonstrate the strong gene-nutrition interaction involving the C677T MTHFR polymorphism. PMID- 11424141 TI - Familial fatal fetal cardiomyopathy with isolated myocardial calcifications: a new syndrome? AB - We describe three male sib fetuses with isolated myocardial calcifications resulting in intrauterine fetal death (IUFD) as early as the second trimester. No evidence for an underlying mitochondrial cytopathy, dystrophinopathy or myopathy was found. There were no signs of inflammation or a metabolic disorder, and the mother had no prenatal exposure of teratogenic drugs. Furthermore, no mutation in the Barth syndrome gene (G4.5) could be detected. Because isolated calcification of the heart and IUFD are not typical of any previously described inherited cardiomyopathy, it may represent a new familial fetal cardiomyopathy. PMID- 11424142 TI - Candidate region for Gilles de la Tourette syndrome at 7q31. AB - Gilles de la Tourette Syndrome (GTS) is a complex neuropsychiatric disorder characterized by motor and vocal tics. The cause of this syndrome is unknown, although based on family studies there is evidence of a strong genetic component. We report on a 13-year-old boy with GTS, minor physical anomalies, and a de novo partial duplication of chromosome 7q [dup(7)(q22.1-q31.1)]. The distal breakpoint in our patient is similar to the breakpoint of an apparently balanced familial translocation t(7;18) segregating with GTS. Together, these cases provide evidence that a gene located in the breakpoint region at 7q31 can be involved in the formation of GTS. PMID- 11424143 TI - Noninvasive prenatal diagnosis of chromosomal aneuploidies by isolation and analysis of fetal cells from maternal blood. AB - The isolation and analysis of nucleated fetal cells (NFCs) from maternal blood may represent a new approach to noninvasive prenatal diagnosis. Although promising, these techniques require highly accurate separation of NFCs from nucleated cells of maternal origin; the two major problems limiting these techniques are the relative rarity of fetal cells in maternal blood and the need to establish their fetal origin. We now report a novel procedure that has allowed accurate separation of NFCs from maternal cells. The technique reported involves direct micromanipulator isolation of histochemically identified hemoglobin F positive nucleated cells to obtain fetal nucleated red blood cells (FNRBCs) of high yield and purity. Using this technique, followed by cell-by-cell multicolor fluorescence in situ hybridization (FISH) analysis of purified FNRBCs, we were able to detect some of the most common human aneuploidies (including Down syndrome, Klinefelter syndrome, and trisomy 13) in 33 pregnant women referred for amniocentesis. The procedure used, which can be completed in <72 hrs, produced complete concordance with the results of amniocentesis. We also confirm findings of prior studies suggesting that the number of FNRBCs in maternal circulation is remarkably higher in abnormal pregnancies than in normal pregnancies, especially in women carrying a fetus with trisomy 21. PMID- 11424144 TI - Bilateral microtia and cleft palate in cousins with Diamond-Blackfan anemia. AB - We report on maternal first cousins with bilateral microtia, micrognathia, cleft palate and hematologic findings of Diamond-Blackfan anemia (DBA). The similarity of findings shared between our cases and a female reported by Hasan and Inoue [1993] suggests that this is a distinctive syndrome, rather than a chance association. DBA is a heterogeneous disorder, caused in about 25% of cases by heterozygous mutations in the RPS19 gene (DBA1). Mutation analysis in our cases did not show an RPS19 mutation, and 2 alleles were present in each. Segregation analysis for DBA1 on chromosome 19 and DBA2 on 8p23 was not consistent with linkage. We conclude that this syndrome of microtia, cleft palate and DBA is not allelic to known DBA loci. PMID- 11424145 TI - Low birth-weight, microcephalic malformation syndrome in a 46,XX girl and her 46,XY sister with agonadism: third report of the Kennerknecht syndrome or autosomal recessive Seckel-like syndrome with previously undescribed genital anomalies. AB - We report on two sisters, one 46,XX with normal female phenotype, the other 46,XY with ambiguous external genitalia and agonadism. Both have a low birth weight and microcephalic malformation syndrome leading to early death. The 46,XX patient also had a diaphragmatic defect. The XY sister, in addition to absence of gonads and Mullerian and Wolffian derivatives, had severe hypoplasia of the pulmonary artery and its branches, multicystic kidneys, and pachygyria. This combination of malformations, in part, fits in the autosomal recessive condition described by Kennerknecht et al. [1993] in a 46,XX and 46,XY pair of sibs with agonadism. The craniofacial appearance of our patients is similar to that observed in the 46,XY sister with agonadism reported by Kennerknecht et al. [1995]. On the other hand, intrauterine growth retardation, microcephaly, and pachygyria are not part of the phenotype of the Kennerknecht "syndrome" and agonadism was not present in our 46,XX patient. We suggest two hypotheses to explain the abnormal phenotype in our cases. The first is that they might represent the third sibship affected by the Kennerknecht syndrome with additional anomalies. Alternatively, our two sibs might have a Seckel or a Seckel-like syndrome. The 46,XY patient would then be the first case of Seckel syndrome with sex reversal and agonadism. PMID- 11424146 TI - Trigonomicrocephaly, severe micrognathia, large ears, atrioventricular septal defect, symmetrical cutaneous syndactyly of hands and feet, and multiple cafe-au lait spots: new acrocraniofacial dysostosis syndrome? AB - We report on a patient with a unique constellation of anomalies comprising trigonomicrocephaly, asymmetric severe micrognathia, large ears, atrioventricular septal defect, vertebral anomalies, bilateral cutaneous syndactyly of fingers and toes, unilateral cryptorchidism and multiple cafe-au-lait spots. The mother of the propositus has multiple cafe-au-lait spots. Search of POSSUM and the London Dysmorphology Database (LDDB) uncovered no similar case. We think that this patient represents a new acrocraniofacial dysostosis syndrome. PMID- 11424147 TI - Comment on Traupe's tribute to Rudolf Happle. PMID- 11424148 TI - Congenital human baculum deficiency: the generative bone of Genesis 2:21-23. PMID- 11424149 TI - Homozygous alpha-thalassemia associated with hypospadias: SEA-type deletion does not affect expression of the -14 gene and loss of the straight theta1-globin gene on 16p13.3 is compensated by its duplicate straight theta2 on chromosome 10. PMID- 11424150 TI - Interface between Robin sequence and ordinary cleft palate. PMID- 11424152 TI - Static magnetic field measurements in residences in relation to resonance hypotheses of interactions between power-frequency magnetic fields and humans. AB - Bowman et al. used epidemiologic data to test a model in which subjects were classified as being "in-resonance" or "not-in-resonance" for 60-Hz magnetic-field exposures depending on single static magnetic-field measurements at the centers of their bedrooms. A second paper by Swanson concluded that a single static magnetic-field measurement is insufficient to meaningfully characterize a residential environment. The main objective of this study was to investigate exposure-related questions raised by these two papers in two U.S. data sets, one containing single spot measurements of static magnetic fields at two locations in homes located in eight states, and the other repeated spot measurements (seven times during the course of one year) of the static magnetic fields at the centers of bedrooms and family rooms and on the surfaces of beds in 51 single-family homes in two metropolitan areas. Using Bowman's criterion, bedrooms were first classified as being in-resonance or not-in-resonance based on the average of repeated measurements of the static magnetic field measured on the bed where the presumed important exposure actually occurred. Bedrooms were then classified a second time using single spot measurements taken at the centers of bedrooms, centers of family rooms, or on the surfaces of beds, as would be done in the typical epidemiologic study. The kappa statistics characterizing the degree of concordance between the first (on-bed averages) and second (spot measurements) methods of assessing resonance status were 0.44, 0.33, and 0.67, respectively. This level of misclassification could significantly affect the results of studies involving the determination of resonance status. PMID- 11424153 TI - Chromosomal aberrations in peripheral lymphocytes of train engine drivers. AB - Studies of Swedish railway employees have indicated that railroad engine drivers have an increased cancer morbidity and incidence of chronic lymphatic leukemia. The drivers are exposed to relatively high magnetic fields (MF), ranging from a few to over a hundred microT. Although the possible genotoxic potential of MF is unclear, some earlier studies have indicated that occupational exposure to MF may increase chromosome aberrations in blood lymphocytes. Since an increased level of chromosomal aberrations has been suggested to predict elevated cancer risk, we performed a cytogenetic analysis on cultured (48 h) peripheral lymphocytes of Swedish train engine drivers. A pilot study of 18 engine drivers indicated a significant difference in the frequency of cells with chromosomal aberrations (gaps included or excluded) in comparison with seven concurrent referents (train dispatchers) and a control group of 16 office workers. The engine drivers had about four times higher frequency of cells with chromosome-type aberrations (excluding gaps) than the office workers (P < 0.01) and the dispatchers (P < 0.05). Seventy-eight percent of the engine drivers showed at least one cell per 100 with chromosome-type aberrations compared with 29% among the dispatchers and 31% among the office workers. In a follow-up study, another 30 engine drivers showed an increase (P < 0.05) in the frequency of cells with chromosome-type aberrations (gaps excluded) as compared with 30 referent policemen. Sixty percent of the engine drivers had one or more cells (per 100 cells) with chromosome-type aberrations compared with 30% among the policemen. In conclusion, the results of the two studies support the hypothesis that exposure to MF at mean intensities of 2-15 microT can induce chromosomal damage. PMID- 11424154 TI - Electrochemical treatment of mouse Ehrlich tumor with direct electric current. AB - Electrochemical treatment of cancer utilizes direct electric current (DEC) to produce direct alterations and chemical changes in tumors. However, the DEC treatment is not established and mechanisms are not well understood. In vivo studies were conducted to evaluate the effectiveness of DEC on animal tumor models. Ehrlich tumors were implanted subcutaneously in sixty male BALB/c mice. When the tumor volumes reached 850 mm(3), four platinum electrodes were inserted into the tumors. DEC of 4 mA was applied for 21 min to the treated group; the total charge was 5 C. The healthy and sick control groups were subjected to the same conditions but without DEC. Hematological and chemical parameters as well as histopathological and peritumoral findings were studied. After the electrochemical therapy it was observed that both tumor volume decrease and necrosis percentage increase were significant in the treated group. Moreover, 24 h after treatment an acute inflammatory response, as well as sodium ion decrease, and potassium ion and spleen weight increase were observed in this group. It was concluded that both electrochemical reactions (fundamentally those in which reactive oxygen species are involved), and immune system stimulation induced by cytotoxic action of the DEC could constitute the most important antitumor mechanisms. PMID- 11424155 TI - Measurement of the individual exposure to 50 and 16 2/3 Hz magnetic fields within the Bavarian population. AB - This study investigates the individual magnetic field exposures at 16 2/3 and 50 Hz of 1952 people, selected from the Bavarian population. Personal flux density meters ("Field Watcher FW2A") were worn by the participants for 24 h. Every second, the flux density was recorded for both frequencies and for the three spatial axes (dynamic range per axis: several nT up to 100 microT at 50 Hz, 150 microT at 16 2/3 Hz). For 50 Hz fields, the mean of the 1,952 individual means was 0.101 microT and that of the individual medians was 0.047 microT. High level exposures occurred mainly during working hours. Only 2.4% of the subjects showed individual medians higher than 0.2 microT. About 53% of all volunteers were working on the day of recording. Levels for craftsmen (n = 148; mean individual mean: 0.166 microT) were generally higher than those for office workers (n = 624; mean individual mean: 0.107 microT). Flux densities exceeding 100 microT at 50 Hz were measured in 31 persons. The total time with such extreme exposures amounts to nearly 21 min, less than 0.001% of the total time for all measurements (5.3 years). To our knowledge, this is the first exposure study where 16 2/3 Hz magnetic fields (caused by electrified railways) have additionally been monitored over 24 h. For persons living next to railway lines, the mean individual mean (0.156 microT) and mean individual median (0.102 microT) were calculated. Over all, the mean exposures are only 0.1% of the magnetic flux density limit for 50 Hz (100 microT) and about 0.05% of the limit (300 microT) for 16 2/3 Hz recommended by the International Commission on Non-Ionizing Radiation Protection. PMID- 11424156 TI - Exposure of Caenorhabditis elegans to extremely low frequency high magnetic fields induces stress responses. AB - Responses of the small heat shock protein gene, hsp-16, were examined in transgenic Caenorhabditis elegans exposed to electromagnetic fields. Expression of the hsp-16-lacZ gene was enhanced when transgenic animals were exposed to magnetic fields up to 0.5 T at 60 Hz. The hsp-16 promoter was more efficiently expressed at the embryonic than at the post-embryonic stage irrespective of exposure. Promoter activity was more sensitive to the stimulus in the intestine at the post-embryonic stage. Evidence is presented that the induction occurs at the transcriptional step of hsp-16. PMID- 11424157 TI - Effects of extremely low frequency magnetic field on fertility of adult male and female rats. AB - To investigate the effects of an extremely low-frequency (ELF) magnetic field on their fertility, adult male and female Sprague-Dawley rats were exposed to a 50 Hz sinusoidal magnetic field of approximately 25 microT (rms) for 90 days before they were mated with unexposed counterparts. Exposure to a 50 Hz field reduced male rat fertility. The number of pregnant females was reduced when mated with exposed males, and the number of resorptions increased. The effects of magnetic field on male fertility were shown to be partly reversible, when the same exposed group of males were remated 45 and 90 days after being removed from the fields. Exposure of adult female rats to 50 Hz magnetic fields for 90 days before mating significantly reduced their fertility. The mean numbers of implantations and living fetuses per litter were statistically significantly decreased in the 50 Hz group. These results suggest that low frequency magnetic fields have some adverse effects on fertility of male and female rats. PMID- 11424158 TI - Effects of 50 Hz magnetic field on cell cycle kinetics and the colony forming ability of budding yeast exposed to ultraviolet radiation. AB - To investigate the effects of extremely low frequency magnetic fields on ultraviolet radiation (UV) exposed budding yeast, haploid yeast (Saccharomyces cerevisiae) cells of the strain SEy2101a were exposed to 50 Hz sine wave magnetic field (MF) of 120 microT with simultaneous exposure to UV radiation. Most of the UV energy was in the UVB range (280-320 nm). The biologically weighted (CIE action spectrum) dose level for the UV radiation was 175 J/m2. We examined whether 50 Hz MF affected the ability of UV irradiated yeast cells to form colonies (Colony Forming Units, CFUs). In addition, the effect of coexposure on cell cycle kinetics was investigated. Although the significant effect of MF on the cell cycle phases of UV exposed yeast cells was seen only at one time point, the overall results showed that MF exposure may influence the cell cycle kinetics at the first cycle after UV irradiation. The effect of our particular MF exposure on the colony forming ability of the UV irradiated yeast cells was statistically significant 420 min after UV irradiation. Moreover, at 240, 360, and 420 min after UV irradiation, there were fewer CFUs in every experiment in (UV+MF) exposed populations than in only UV exposed yeast populations. These results could indicate that MF exposure in conjunction with UV may have some effects on yeast cell survival or growth. PMID- 11424159 TI - Extended exposure of adult and fetal mice to 50 Hz magnetic field does not increase the incidence of micronuclei in erythrocytes. AB - The flow cytometer-based micronucleus assay was used to study the effects on chromosomes in erythroid cells of CBA/Ca mice after extended exposure to 50 Hz magnetic field (MF), 14 microT, peak-to-peak (p-p). The study included two different experiments: (a) mice exposed in utero during 18 days of their prenatal stage, and (b) adult mice exposed for 18 days. In experiment (a) 35 days after exposure was terminated, peripheral blood was drawn from the mice exposed in utero to determine whether the exposure had a genotoxic effect on the pluripotent erythroid stem cells. About 200000 polychromatic erythrocytes (PCE) and 200000 normochromatic erythrocytes (NCE) were analysed from each of 20 exposed mice. The EMF exposure did not significantly change the frequency of micronucleated PCE or NCE in comparison with 20 sham-irradiated mice. There was no difference in the proportion of PCE between exposed and unexposed animals. Similarly, in experiment (b) no differences were seen between EMF exposed and unexposed adult mice when samples of peripheral blood were taken at the end of exposure and analyzed for micronuclei in PCE and NCE. The proportion of PCE was the same in both groups. The results indicate that exposure to EMF does not induce direct or indirect effects on chromosomes in erythroid cells expressed as increased levels of micronucleated erythrocytes of mice. No indications of delayed genetic effects were found. PMID- 11424160 TI - Reactions of keratinocytes to in vitro millimeter wave exposure. AB - The effects of millimeter waves (MW) on human keratinocytes were studied in vitro using the HaCaT keratinocyte cell line. MW-induced modulation of keratinocyte function was studied in proliferation, adhesion, chemotaxis, and interleukin 1beta (IL-1beta) production assays. Spontaneous proliferation, adhesion to tissue culture plate, random migration, and IL-8- and RANTES induced chemotaxis were not affected by exposure of cells to millimeter waves under the following conditions: frequency, 61.22 GHz; SAR, 770 W/kg; duration of exposure, 15-30 min. However, MW irradiation resulted in a modest but statistically significant increase in the intracellular level of IL-1beta. These data suggest that exposure of human skin (with keratinocytes being the major component of epidermis) to MW can cause activation of basal keratinocytes resulting in an elevated level of IL-1beta production. PMID- 11424161 TI - Effect of a permanent magnetic field on wing size parameters in Drosophila melanogaster. AB - The width and length variability of both D. melanogaster wings were measured in samples of flies after two and six generations under a permanent magnetic field of 35 mT. While in earlier generations under exposure sexes differ in the size response, later they both show a decrease in the wing size under the magnetic field exposure if compared to the control. The bilateral asymmetry in wings as a potential indicator of developmental instability does not increase significantly under exposure. The ecological and adaptive implications of the change of the magnetic intensity as an environmental factor generating stress in populations is discussed. PMID- 11424163 TI - Interventions for depressed elderly primary care patients. PMID- 11424164 TI - The systematic assessment of depressed elderly primary care patients. AB - Studies of the primary care treatment of depressed elderly patients are constrained by limited time and space and by subject burden. Research assessments must balance these constraints with the need for obtaining clinically meaningful information. Due to the wide-ranging impact of depression, assessments should also focus on suicidality, hopelessness, substance abuse, anxiety, cognitive functioning, medical comorbidity, functional disability, social support, personality, service use and satisfaction with services. This paper describes considerations concerning the assessment selection process for primary care studies, using the PROSPECT (Prevention of Suicide in Primary Care Elderly: Collaborative Trial) study as an example. Strategies are discussed for ensuring that data are complete, valid and reliable. PMID- 11424165 TI - The benefits of a broader perspective in case-finding for disease management of depression: early lessons from the PROSPECT Study. AB - The PROSPECT (Prevention of Suicide in Primary Care Elderly - Collaborative Trial) Study evaluates collaborative care for the treatment of current depressive disorders in older primary care patients. Using screening data, we evaluate implications of expanding the focus to additional patients who report taking psychotropic medication or having a history of depression. Some 13.8% of the patients screened were taking a psychotropic medication and 7.3% were both taking an antidepressant and had a Center for Epidemiologic Studies - Depression Scale (CES-D) score of > 15. Patients who reported current antidepressant use and those with a history of depression also had elevated CES-D scores. Conversely, most patients having an elevated CES-D score also reported taking an antidepressant or having a history of depression. These latter criteria for potential caseness yielded a larger number of patients than those identified by the CES-D alone. More comprehensive approaches to the management of depression should attend to patients already taking psychotropic medication and symptomatic patients with a history of depression, as well as those with current syndromal depression. PMID- 11424166 TI - Managing late-life depression in primary care practice: a case study of the Health Specialist's role. AB - Complexities in the diagnosis and treatment of late-life depression have stimulated various strategies for assisting the primary care physician to fulfil these tasks more effectively. The role of Health Specialist was developed for this purpose in a study to reduce suicidality among older depressed patients. This role includes clinical and case management tasks which aim to provide the physician with timely, patient-specific information and recommendations. Evolution of this role and its rewards/stressors during the study's first year are described. PMID- 11424167 TI - Pharmacological treatment of depression in older primary care patients: the PROSPECT algorithm. AB - PROSPECT (Prevention of Suicide in Primary care Elderly-Collaborative Trial) is testing whether a trained clinician (the 'health specialist') can work in close collaboration with a primary care physician to implement a comprehensive depression management program and improve outcomes in older depressed patients. An algorithm guiding the selection and use of antidepressant medications has been developed to assist PROSPECT health specialists. This algorithm is presented and the rationale underlying the proposed treatment sequence is discussed. The PROSPECT algorithm builds upon existing guidelines after updating them and adapting them to the special circumstances of older primary care patients. Special attention has been paid to the tolerability and the target doses of the recommended antidepressant agents and to the duration of antidepressant trials. Patients who are unable to tolerate or do not respond to an antidepressant can be switched to another agent or be treated with interpersonal psychotherapy. Agents that produce only a partial response can be combined with other antidepressants or with interpersonal psychotherapy. Treatments for which empirical evidence exists are favored. However, treatments that are often poorly tolerated by elderly patients are given lower priority than treatments more likely to be tolerated. Similarly, trials that are simpler to implement in primary care are favored. PMID- 11424168 TI - High-risk management guidelines for elderly suicidal patients in primary care settings. AB - Older adults, especially older white men, are more likely to commit suicide than other age-groups. The assessment and management of suicide ideation and behavior for the elderly is especially relevant for primary care physicians because many patients visit their physician shortly before committing suicide. In 1993, the Agency for Health Care Policy and Research (AHCPR) released clinical practice guidelines for the treatment of depression in primary care settings. The AHCPR guidelines offered some assistance with respect to detecting and treating patients who are suicidal. However, these guidelines lacked detailed and specific instructions for managing high-risk patients within the context of intervention research. The current set of guidelines builds upon this previous work by describing (1) a general approach for interacting with suicidal patients; (2) structured assessments for determining the degree of risk for suicide; (3) crisis intervention strategies; and (4) ongoing management procedures for working with suicidal patients in primary care settings. PMID- 11424169 TI - Treatment as usual (TAU) control practices in the PROSPECT Study: managing the interaction and tension between research design and ethics. AB - The use of treatment as usual (TAU) as a control condition may pose the considerable challenge of maintaining both scientific rigor and meeting high ethical standards in experiments on human subjects. The authors illustrate the tension and explore the relationship between research design and ethics, especially the interaction between the two, in the NIMH-funded PROSPECT study (Prevention of Suicide in Primary Care Elderly - Collaborative Trial). The goal of PROSPECT is to determine whether placement of a depression health specialist in primary care practices will have a favorable impact on rates of depression, hopelessness and suicidal ideation in elderly primary care patients with major or persistent minor depression. PROSPECT randomly assigns practices either to an intervention arm (which includes the provision of depression health specialists) or to an enhanced care arm (TAU, with the addition of screening and assessment services). TAU, enhanced by the provision of screening and assessment services, is to be used as a benchmark for measuring the effectiveness of PROSPECT's intervention. However, TAU in the epidemiological and clinical literature has also been linked to high rates of suicide in the elderly related to unrecognized and untreated or under-treated depression. The authors present their approach to managing the tension, or interaction, between the use of TAU for scientific and public health purposes and the requirement for beneficence, that is, the duty to assure the safety of human subjects in research and to do no harm. Through enhancements of TAU, by the provision of information to primary care physicians concerning the psychiatric status of their patients, the investigators attempt to meet the challenge of maintaining rigor and meeting high ethical standards. PMID- 11424170 TI - Improvement of cognitive functions after cobalamin/folate supplementation in elderly patients with dementia and elevated plasma homocysteine. AB - OBJECTIVES: To investigate the effect of cobalamin/folate supplementation on cognitive function in elderly patients with dementia. METHOD: The cobalamin/folate status of the patients was evaluated by measuring plasma homocysteine, serum methylmalonic acid, serum cobalamin and blood folate. Thirty three patients were studied and repeatedly assessed with the Mini-Mental State Examination (MMSE) and 'A short cognitive performance test for assessing memory and attention' (SKT) during vitamin substitution. RESULTS: Patients with mild moderate dementia and elevated plasma homocysteine levels improved clinically with increased test scores after vitamin substitution, while severely demented patients and patients with normal plasma homocysteine levels did not improve clinically. CONCLUSIONS: Plasma homocysteine may be the best marker for detecting treatable cobalamin/folate deficiency in patients with dementia. PMID- 11424171 TI - Assessment of dementia in nursing home residents by nurses and assistants: criteria validity and determinants. AB - OBJECTIVES: To describe the criterion validity of nursing home staff's assessment of organic disorder compared with ICD-10 criteria, and to identify determinants of staff assessment of organic disorder. METHOD: Two hundred and eighty-eight residents were diagnosed using the GMS-AGECAT. Nursing staff members were interviewed about the residents' activities of Daily Living, behavioural problems, orientation in surroundings and communication skills, and asked if the resident had an organic disorder. Multiple logistic regression was used to select the items that most strongly determined staff assessment of organic disorder. RESULTS: Sixty-two per cent of the residents were diagnosed by GMS-AGECAT as having organic disorder, 78% of these were correctly identified by the staff. Whether analysed among residents with or without organic disorder, or in the total group of residents, the staff assessment of the presence of organic disorder depended on a limited set of behavioural characteristics of the resident, namely 'going to the toilet in inappropriate places', 'saying things that do not make sense' and impairment in orientation. CONCLUSIONS: Staff comprehension of organic disorder resulted in over- as well as under-labelling of residents, a tendency that will affect communication with medical personnel and may lead to inadequate or wrong medical treatment and to negative performance as well as negative role expectations in everyday life in nursing homes. PMID- 11424172 TI - Depression, depressive symptoms and mortality in persons aged 65 and over living in the community: a systematic review of the literature. AB - BACKGROUND: No recent attempt has been made to synthesize information on mortality and depression despite the theoretical and practical interest in the topic. Our objective was to estimate in the older population the influence on mortality of depression and depressive symptoms. METHODS: Data sources were: Medline, Embase, personal files and colleagues' records. Studies were considered if they included a majority of persons aged > or = 65 years at baseline either drawn from a total community sample or drawn from a random sample from the community. Samples from healthcare facilities were excluded. Effect sizes were extracted from the papers; if they were not included in the published papers, effect sizes were calculated if possible. No attempt was made to contact authors for missing data. RESULTS: We found 21 reports on 23 cohorts using depression diagnosis. For 15 of these, odds ratios were pooled using the Greenland method based on confidence intervals (CIs), giving an estimated odds ratio for mortality with depression of 1.73 (95% CI 1.53 to 1.95). A fixed effects meta-regression of these studies suggested that longer follow-up predicted smaller effect sizes (log odds ratios -0.096 per year (95% CI -0.179 to -0.014)). There is a weak suggestion of a reduced effect of depression on mortality for women. We were unable to pool effect sizes from the 17 studies using symptom totals and scales, or from eight studies of specific symptoms. CONCLUSIONS: The studies show that diagnosed depression in community-resident older people is associated with increased mortality. The picture for sex differences is still unclear. PMID- 11424173 TI - Cognitive impairment and functional status in elderly institutionalized patients with schizophrenia. AB - OBJECTIVE: The relationship of cognitive impairment to functional status in older adults with schizophrenia was investigated. PATIENTS: Ninety-three psychiatric inpatients with schizophrenia between the ages of 65 and 88 years. Two subsets of this sample, consisting of 48 and 24 patients, were studied with a greater number of assessment instruments. MEASURES: The Mini-Mental State Examination (MMSE) was used for brief assessment of overall cognitive functioning, and the Psychogeriatric Dependency Rating Scale (PGDRS) was administered to assess functional status. The cognitive test battery from the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) and/or an expanded neuropsychological battery, was given to a subset of the patients. RESULTS: In the overall sample, patients with greater global cognitive impairment had higher levels of rated impairment on the individual items that comprised the Orientation and Physical, but not Behavior, subscales of the PGDRS. Furthermore, in the two subsamples, specific neuropsychological measures of problem-solving, word list learning, naming and constructional praxis were related to overall measures of outcome. CONCLUSIONS: Neuropsychological deficit and psychosocial outcome are multi dimensional entities that relate to one another in complex ways. PMID- 11424174 TI - Dietary and endogenous antioxidants in dementia. PMID- 11424175 TI - Definition of 'neuroleptic sensitivity'. PMID- 11424176 TI - What is the impact of a psychogeriatric admission on the prescription of psychotropic and non-psychotropic drugs? PMID- 11424177 TI - Role of glutathione S-transferase mu (GSTM1) in styrene-7,8-oxide toxicity and mutagenicity. AB - In the human glutathione S-transferase (GST) mu gene family, homozygous deletion of GSTM1 is the null phenotype (frequency of approximately 50% in Caucasians). In the current study, GSTM1 status was determined in human cell lines using reverse transcriptase, polymerase chain reaction, and immunochemistry. Cell lines were challenged with a range of doses of styrene-7,8-oxide (SO) and then toxicity and genotoxicity were monitored. Toxicity was determined by growth in flasks and genotoxicity by cloning in microplates in the presence/absence of 6-thioguanine, to detect mutations at the hypoxanthine phosphoribosyltransferase (hprt) locus. A SO concentration-dependent decrease in survival was observed for all cell lines, with GSTM1-deficient lines being more sensitive. The IC(50)s of deficient and proficient cell lines were 0.45 and 0.55 mM SO, respectively. The difference between survival of GSTM1-deficient and -proficient cell lines approached statistical significance. The background mutation frequency of GSTM1-deficient cell lines was 2 x 10(-5), and that of GSTM1-proficient cell lines was 3 x 10( 6). GSTM1-deficient cell lines were significantly more sensitive than GSTM1 proficient cell lines to mutation induction for concentrations up to 2.5 mM SO (P < 0.001, regression analysis). These results suggest that cell lines containing metabolically competent GSTM1 are able to efficiently use GSTM1 to conjugate SO and reduce its hazard. This supports the epidemiological evidence that GSTM1 influences sensitivity to chemical carcinogenesis and subsequent risk of cancer induction. PMID- 11424178 TI - Just how does the cII selection system work in Muta Mouse? AB - The lambda CII protein is an essential component in the lytic vs. lysogeny decision a bacteriophage makes upon infection of a host at low temperatures. The protein interacts with numerous phage promoters modulating the expression of the CI repressor, thus providing the mechanism for lysogenization soon after infection. The Big Blue and Muta Mouse are two widely used in vivo mutational model systems. The assays rely on retrievable lambda-based transgenes housing mutational targets (lacI or lacZ, respectively). The transgenes provide an elegant vehicle for the quantification of mutations sustained in virtually any tissue of the rodent. The use of the bacteriophage cII locus as an alternative, or additional mutational target for use with the Big Blue rodent system was first reported by Jakubczak et al. ([1996]: Proc Natl Acad Sci USA 93:9073-9078). More recently, this selection assay has been applied successfully to the Muta Mouse (Swiger et al. [1999]: Environ Mol Mutagen 33:201-207). The use of an Hfl bacterial strain and low temperature allows the determination of mutations sustained at the cII locus in either system, with high fidelity. The cII selection assay in the Big Blue relies on the presence of the lambda repressor protein CI. In contrast, the recombinant construct used to make the Muta Mouse transgene lacks functional CI protein. Nevertheless, we report an excellent system for quantifying mutations at the cII locus in Muta Mouse. Just how does cII selection work in the Muta Mouse? Written in the context of lambda recombinant genetics, this paper explores the question further. PMID- 11424179 TI - Further characterization and validation of gpt delta transgenic mice for quantifying somatic mutations in vivo. AB - The utility of any mutation assay depends on its characteristics, which are best discovered using model mutagens. To this end, we report further on the characteristics of the lambda-based gpt delta transgenic assay first described by Nohmi et al. ([1996]: Environ Mol Mutagen 28:465-470). Our studies show that the gpt transgene responds similarly to other transgenic loci, specifically lacZ and cII, after treatment with acute doses of N-ethyl-N-nitrosourea (ENU). Because genetic neutrality is an important factor in the design of treatment protocols for mutagenicity testing, as well as for valid comparisons between different tissues and treatments, a time-course study was conducted. The results indicate that the gpt transgene, like cII and lacZ, is genetically neutral in vivo. The sensitivities of the loci are also equivalent, as evidenced by spontaneous mutant frequency data and dose- response curves after acute treatment with 50, 150, or 250 mg/kg ENU. The results are interesting in light of transgenic target size and location and of host genetic background differences. Based on these studies, protocols developed for other transgenic assays should be suitable for the gpt delta. Additionally, a comparison of the gpt and an endogenous locus, Dlb-1, within the small intestine of chronically treated animals (94 microg/mL ENU in drinking water daily) shows differential accumulation of mutations at the loci during chronic exposure. The results further support the existence of preferential repair at endogenous, expressed genes relative to transgenes. PMID- 11424180 TI - Gender difference in DNA adduct levels among nonsmoking lung cancer patients. AB - Lung cancer is the leading cause of cancer mortality in Taiwanese women. Cigarette smoking cannot explain the high lung cancer mortality in this population because less than 10% of women in Taiwan are smokers. Therefore, environmental factors other than smoking may play an important role in lung cancer development in female nonsmokers. The purpose of this study was to elucidate the role of environmental carcinogen exposure in lung cancer development in Taiwanese female nonsmokers, based on DNA adduct formation. We collected nontumorous lung tissues resected from 62 nonsmoking lung cancer patients and 20 noncancer controls to investigate whether differences in susceptibility to DNA adduct formation exist between men and women. (32)P postlabeling and ELISA (enzyme-linked immunosorbent assay) with polyclonal antibody against BPDE (7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10 tetrahydrobenzo[a]pyrene)-DNA adduct were used to evaluate DNA adduct levels in lung tissues of study subjects. Our data showed that the DNA adduct levels of lung cancer patients determined by both assays were significantly higher than those of noncancer controls (P = 0.0001 for (32)P-postlabeling; P = 0.01 for ELISA). Moreover, DNA adduct levels in females were markedly greater than those in males (P = 0.014 for (32)P-postlabeling; P = 0.001 for ELISA). The difference in DNA adduct levels could not be explained by genetic polymorphisms of cytochrome P-4501A1 (CYP1A1) or glutathione S-transferase (GSTM1), as determined by polymerase chain reaction and restriction fragment length polymorphism. These results demonstrate that lung cancer patients have a higher susceptibility to DNA damage than that of noncancer controls. In addition, differences in susceptibility to DNA damage derived from environmental carcinogen exposure were observed between male and female nonsmokers. In conclusion, high susceptibility to DNA damage in females may partially explain the high mortality rate of lung cancer in nonsmoking Taiwanese women. PMID- 11424181 TI - Spontaneous microdeletions and microinsertions in a transgenic mouse mutation detection system: analysis of age, tissue, and sequence specificity. AB - A total of 3497 independent spontaneous mutations were examined using the Big Blue transgenic mouse mutation detection system. Base substitutions predominate, although 16% of somatic and germline mutations are microdeletions, microinsertions, or deletions combined with insertions. The pattern of microdeletions and microinsertions is similar in both the lacI transgene and the human p53 gene. Single-base deletions (D1) and insertions (I1) are evenly distributed in the lacI transgene, whereas microdeletions from 2 to 50 bp are clustered at two regions (bp 129-228 and 529-628). The pattern of microdeletions and microinsertions is similar between young (< or =3 months) and old (25 months) mice. Brain tissue has a paucity of deletions combined with insertions when compared with that of thymus and nine other tissues (P = 0.01). A 16-bp deletion at lacI base position 272 is a tissue-specific hotspot preferentially occurring in brain. Approximately 68 and 93% of D1 and I1, respectively, occur at mononucleotide repeats. The frequencies of D1 and I1 in mononucleotide repeats increase in an exponential manner with the length of the repeat. The lacI transgene shows similarity to the human p53 gene in the pattern of microdeletions and microinsertions and the size distribution of microdeletions. PMID- 11424182 TI - Mutagenicity of 2-alkylpropenals in Salmonella typhimurium strain TA 100: structural influences. AB - alpha,beta-Unsaturated aldehydes are a class of mutagenic and carcinogenic compounds that form promutagenic 1,N(2)-propanodeoxyguanosine adducts. They are important industrial and environmental compounds, are formed endogenously, and are found in food. We recently published structure-mutagenicity relationships for 3-alkyl substituted alpha,beta-unsaturated aldehydes (beta-alkylacroleins) and here we present structural influences on the mutagenicity of the 2-alkyl substituted alpha,beta-unsaturated aldehydes (alpha-alkylacroleins), 2 methylacrolein, 2-ethylacrolein, 2-propylacrolein, and 2-butylacrolein, in Salmonella typhimurium TA 100. All four alkylacroleins are mutagenic without S9 mix; however, the results are strongly influenced by bacterial toxicity of the alkylacroleins. In general, toxicity increases with increasing length of the alkyl substituent. The increasing toxicity with increasing alkyl groups can be explained by increasing lipophilicity that allows the compounds to better penetrate into the bacterial cell. Other structural effects, such as steric hindrance of the deoxyguanosine binding (DNA-adduct formation) and the positive inductive effect of the alkyl groups, have only a slight effect on mutagenesis. Addition of S9-mix leads to an increase in the absolute revertant peak values but a decrease in mutagenic activities, as expressed by revertants per micromol. This effect is also observed with heat-inactivated S9-mix and does not depend on metabolic activation. The effect of S9-mix can be explained by partial detoxication of the substances by nucleophilic components of the S9-mix such as glutathione. PMID- 11424183 TI - XPA protein alters the specificity of ultraviolet light-induced mutagenesis in vitro. AB - Studies of ultraviolet (UV) light mutagenesis have demonstrated mutations at common sites in the target genes of shuttle vector plasmids replicated in cultured cells or by cellular extracts. The reasons for the specific pattern of mutagenesis are largely unknown. We have examined the specificity of UV-induced mutagenesis by replicating plasmid pLS189, irradiated with 40 J/m(2) UVC or unirradiated, in either xeroderma pigmentosum group A (XP-A) or HeLa cellular extracts. The XP-A extract displayed slightly lower replication ability, but produced a higher mutant frequency, compared to that of HeLa extract. Use of irradiated plasmid inhibited replication by an average of 63% and increased the mutant frequency by an average of 16.7-fold. Analysis of mutation spectra revealed nonrandom patterns of mutagenesis that differed significantly between HeLa and XP-A extracts. In comparison to HeLa extract, replication in XP-A extract resulted in lower frequencies of GC --> AT transitions and tandem double base substitutions, and a higher frequency of deletions. Replication in HeLa extract produced hotspots at positions 100, 108, and 156 that were not produced by XP-A extract. Furthermore, XP-A extract produced hotspots at positions 124, 133, and 164, sites not characteristic of previous UV-induced mutagenesis studies using XPA-expressing cells. Addition of purified XPA protein to reactions containing XP-A extract altered each of these parameters, including loss of the hotspots at positions 124 and 133, to yield a more HeLa-like spectrum. These results indicate a previously uncharacterized role of the XPA protein in influencing the specificity of UV-induced mutagenesis during DNA replication. PMID- 11424184 TI - Effect of mutagen-induced cell lethality on the dose response of germline mutations. AB - Molecular tests for mutations require a sample of tissue from which DNA is extracted, to determine the presence or absence of one or more mutations per sample. To ensure mutation fixation each sample must consist of an equal number of cells that have had one or more DNA replications. In an in vivo test, surviving stem cells compensate to give the same number of cells per sample, leaving as the only evidence for stem cell lethality the increase in mutants of clonal origin because the mutant clone developed from a population of fewer stem cells. A problem is that an increase in mutagen dose increases stem cell death, resulting in a decreased number of surviving target cells, thus giving a downward bias of samples with one or more mutations per sample. To compare in vivo tests with molecular tests we will use as a model system the sex-linked recessive lethal (SLRL) test for germ cell mutations in Drosophila melanogaster. Spermatogonia cells in male larvae were exposed to ENU and mutations detected in sperm cells from adults. The same SLRL data were analyzed by two methods: (1) The conventional analysis of SLRL data, in which each mutation of a cluster of mutations of common origin was counted. (2) An analysis was used to simulate a sample for molecular analysis by determining mutations per male with an equal size sample of progeny per male. With this second analysis a correction factor is required based on the change in cluster size of mutants of common origin. PMID- 11424185 TI - Direct separation of in vivo and in vitro am3 revertants in transgenic mice carrying the phiX174 am3, cs70 vector. AB - Target genes in most transgenic systems have higher spontaneous mutation frequencies than do endogenous mammalian genes. Spontaneous mutations in transgenes predominantly arise from three sources: (1) mutations fixed in the animals, (2) mutations arising from replication errors caused by damage to the DNA that may have occurred in vivo or in vitro and then was fixed during amplification of the vector in vitro, and (3) mutations arising during replication of non-revertant phages in non-permissive bacteria. An assay based on single bursts was developed to directly distinguish between the in vivo and in vitro origins of revertants. The size of the aliquot is determined by mutant frequency and is adjusted so that ideally no more than 10 to 20% of the aliquots contain a bacterial cell transformed with a mutant phage. Mutations are detected as revertants of an amber mutation (am3) in phiX174 am3, cs70. The minimum burst size of non-revertant phiX am3, cs70 from splenic DNA on a permissive bacterial strain was larger than 30 plaque-forming units (pfu). Based on this observation, a burst size of 31 plaque-forming revertants was chosen as the minimum burst size of a fixed mutation. The single burst assay was tested on DNA from spleens of animals that were treated with 150 mg/kg 1-ethyl-1 nitrosurea. Only the fraction of aliquots with single bursts of revertants (> 30) increased in the treated animals compared to the controls. In contrast, there was no difference between treated and control animals for revertant frequencies calculated for burst sizes < or =30 pfu. Among the spontaneous mutations, only 30% were caused by mutations fixed in animals (i.e. burst size >30 pfu). Total average revertant frequency measured in DNA from treated animals was less than twofold more than the average spontaneous frequency (P = 0.048). When frequencies were based on burst sizes >30, there was a 4.6-fold increase among treated animals compared with controls (P = 0.026). The single burst-assay resulted in a more sensitive test for mutagenicity because it eliminated noise from in-vitro mutations. PMID- 11424186 TI - Estimation of the average burst size of Phix174 am3, cs70 for use in mutation assays with transgenic mice. AB - In mutation assays using transgenic mice, with recoverable vectors such as PhiX174 am3, cs70, mutations originate from two sources: (1) in vivo mutations, that is, mutations that were fixed in the mouse, or (2) ex vivo mutations, that is, mutations that were fixed during recovery or plating. When a bacteriophage infects a bacterium, it multiplies and bursts the cell, releasing a number of phages referred to as the burst size. Our method for distinguishing between in vivo mutations and ex vivo mutations estimates the average number of bursts, the denominator of in vivo mutant frequencies, by dividing the total plaque-forming units (PFU) by the average number of phages in a burst. Herein, we outline a probability model relating observed plaque counts to the burst size and present the statistical method used to estimate the burst size. The average size of a single burst from nonrevertant phages was estimated in eight studies under the conditions of our mutation assay. The average burst size was stable across studies at 182.5 plaques per burst (standard error, 14.25). The probability that a burst is a specific size was approximated by a negative binomial distribution, which implies a Poisson-Pascal distribution for the observed plaque counts. The observed plaque counts were adequately fit by this approximation. Environ. Mol. Mutagen. 37:356-360, 2001 Published 2001 Wiley-Liss, Inc. PMID- 11424187 TI - Electrical coupling between glial cells in the rat retina. AB - The strength of electrical coupling between retinal glial cells was quantified with simultaneous whole-cell current-clamp recordings from astrocyte-astrocyte, astrocyte-Muller cell, and Muller cell-Muller cell pairs in the acutely isolated rat retina. Experimental results were fit and space constants determined using a resistive model of the glial cell network that assumed a homogeneous two dimensional glial syncytium. The effective space constant (the distance from the point of stimulation to where the voltage falls to 1/e) equaled 12.9, 6.2, and 3.7 microm, respectively for astrocyte-astrocyte, astrocyte-Muller cell, and Muller cell-Muller cell coupling. The addition of 1 mM Ba(2+) had little effect on network space constants, while 0.5 mM octanol shortened the space constants to 4.7, 4.4, and 2.6 microm for the three types of coupling. For a given distance separating cell pairs, the strength of coupling showed considerable variability. This variability in coupling strength was reproduced accurately by a second resistive model of the glial cell network (incorporating discrete astrocytes spaced at varying distances from each other), demonstrating that the variability was an intrinsic property of the glial cell network. Coupling between glial cells in the retina may permit the intercellular spread of ions and small molecules, including messengers mediating Ca(2+) wave propagation, but it is too weak to carry significant K(+) spatial buffer currents. PMID- 11424188 TI - Pigment epithelium-derived factor supports normal Muller cell development and glutamine synthetase expression after removal of the retinal pigment epithelium. AB - In conditions in which the retinal pigment epithelium (RPE) is dystrophic, carries a genetic mutation, or is removed physically, Muller cells undergo degenerative changes that contribute to the retinal pathology. We previously demonstrated that pigment epithelium-derived factor (PEDF), a glycoprotein secreted by the RPE cells with neuroprotective and differentiation properties, protects against photoreceptor degeneration induced by RPE removal. The purpose of the present study was to analyze the putative gliosupportive activity of PEDF on Muller cells of RPE-deprived retinas and assess whether protection of Muller cells was correlated with improved photoreceptor outer segment assembly. Eyes were dissected from Xenopus laevis tadpoles, and the RPE was removed before culturing in medium containing purified PEDF, PEDF plus anti-PEDF, or medium alone. Control eyes matured with an adherent RPE or in medium containing PEDF plus nonimmune serum. Muller cell ultrastructure was examined. Glial fibrillary acidic protein (GFAP) and glutamine synthetase were localized immunocytochemically, and the corresponding protein levels were quantified. In control retinas, Muller cells were structurally intact and formed adherens junctions with neighboring photoreceptors. In addition, they did not express GFAP, whereas glutamine synthetase expression was high. RPE removal dramatically altered the ultrastructure and biosynthetic activity of Muller cells; Muller cells failed to form adherens junctions with photoreceptors and glutamine synthetase expression was suppressed. PEDF prevented the degenerative glial response; Muller cells were ultrastructurally normal and formed junctional complexes with photoreceptors. PEDF also preserved the expression of glutamine synthetase at near-normal levels. The morphogenetic effects of PEDF were blocked by the anti-PEDF antibody. Our study documents the glioprotective effects of PEDF and suggests that maintenance of the proper Muller cell ultrastructure and expression of glutamine synthetase may be necessary to support the proper assembly of photoreceptor outer segments. PMID- 11424189 TI - Transplantation of an acutely isolated bone marrow fraction repairs demyelinated adult rat spinal cord axons. AB - The potential of bone marrow cells to differentiate into myelin-forming cells and to repair the demyelinated rat spinal cord in vivo was studied using cell transplantation techniques. The dorsal funiculus of the spinal cord was demyelinated by x-irradiation treatment, followed by microinjection of ethidium bromide. Suspensions of a bone marrow cell fraction acutely isolated from femoral bones in LacZ transgenic mice were prepared by centrifugation on a density gradient (Ficoll-Paque) to remove erythrocytes, platelets, and debris. The isolated cell fraction contained hematopoietic and nonhematopoietic stem and precursor cells and lymphocytes. The cells were transplanted into the demyelinated dorsal column lesions of immunosuppressed rats. An intense blue beta galactosidase reaction was observed in the transplantation zone. The genetically labeled bone marrow cells remyelinated the spinal cord with predominately a peripheral pattern of myelination reminiscent of Schwann cell myelination. Transplantation of CD34(+) hematopoietic stem cells survived in the lesion, but did not form myelin. These results indicate that bone marrow cells can differentiate in vivo into myelin-forming cells and repair demyelinated CNS. PMID- 11424190 TI - Role of the extracellular matrix in myelination of peripheral nerve. AB - Assembly of the extracellular matrix (ECM) has been tightly linked to compact myelin formation in the peripheral nervous system. We recently demonstrated that myelination of dorsal root ganglion (DRG) axons by Schwann cells may occur in the absence of basal lamina. We have now determined whether laminin deposition occurs around myelinating SC, even though basal lamina has not been assembled. DRG/SC co cultures were prepared from E15 rat embryos and incubated in fully defined medium (B27) with and without ascorbic acid for 21-24 days. Cultures were stained with a rabbit anti-laminin antibody and examined by laser confocal fluorescence microscopy. Myelination occurred in both groups. In the presence of ascorbic acid, there was dense even laminin staining around myelinating SC. In the absence of ascorbic acid, laminin staining was also present but was irregular and less dense. DRG and SC were co-cultured without ascorbic acid in the presence or absence of a function blocking anti-beta(1) integrin receptor antibody. The antibody completely inhibited myelination. Finally, DRG/SC co-cultures were prepared both with and without ascorbic acid and incubated under control conditions or in the presence of continual, gentle motion. Movement in the absence of ECM significantly inhibited myelination. This demonstrates that laminin deposition on the surface of SC but not ECM assembly is required for formation of compact myelin. ECM is required to provide mechanical stability during the process of myelination. PMID- 11424191 TI - Activation of murine cytomegalovirus immediate-early promoter in cerebral ventricular zone and glial progenitor cells in transgenic mice. AB - Cytomegalovirus (CMV) is the most common infectious cause of congenital anomalies of the CNS in humans. We recently reported that the murine cytomegalovirus (MCMV) immediate-early (IE) gene promoter directs astrocyte-specific expression in adult transgenic mice. In the present study, we analyzed the activation of the MCMV IE promoter in developing transgenic mouse brains and compared the activation with that of the Musashi 1 (Msi1) gene, which is expressed in neural progenitor cells, including neural stem cells. During the early phase of neurogenesis, the transgene was expressed predominantly in endothelial cells of the vessels, but not in neuroepithelial cells in which Msi1 was expressed. During later stages of gestation, expression of the transgene was largely restricted to the ventricular zone (VZ) in the CNS, similar to the expression of Msi1. In neurosphere cultures from transgenic embryos in the late phase of neurogenesis, the transgene was expressed in some cells of neurospheres expressing Msi1 and nestin. In neural precursor cells induced to differentiate from stem cells, expression of the transgene was detected in glial progenitor cells, expressing GFAP, nestin, and Msi1, but not in cells expressing MAP2 or MAG. In postnatal development, persistent expression of the transgene was observed in astrocyte lineage cells as was Msi1. These spatiotemporal changes of the MCMV IE promoter activity during development of transgenic mice correlated with susceptible sites in congenital HCMV infection. Moreover, this transgenic mouse model may provide useful model for analysis of the regulation of the switching of neuronal and astrocyte differentiation, and the maintenance of the astrocyte lineage. PMID- 11424192 TI - Characterization of a novel brain-derived microglial cell line isolated from neonatal rat brain. AB - We observed highly aggressively proliferating immortalized (HAPI) cells growing in cultures that had been enriched for microglia. The cells were initially obtained from mixed glial cultures prepared from 3-day-old rat brains. HAPI cells are typically round with few or no processes when cultured in 10% serum containing medium. As the percentage of serum in the medium is decreased, the HAPI cells have more processes. HAPI cells stain for the isolectin B4, OX-42, and GLUT5, which are markers for microglial cells, but the cells do not immunolabel with A2B5, a marker of cells in the oligodendroglial cell lineage, or with the astrocyte-specific marker, glial fibrillary aciidic protein (GFAP). In addition, HAPI cells are capable of phagocytosis. We conclude that HAPI cells are of microglia/macrophage lineage. Exposing HAPI cells to lipopolysaccharide (LPS) induces the mRNAs for tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase (iNOS). LPS exposure also induces secretion of TNF-alpha and production of nitric oxide (NO) in HAPI cells. Because activation of microglia is associated with an increase in iron accumulation and ferritin expression, we tested the hypothesis that iron status affects the production of TNF-alpha and NO. Our studies demonstrate that both iron chelation and iron loading diminished the LPS-induced effect of TNF-alpha and NO. The results of this study indicate that HAPI cells possess the characteristics of microglia/brain macrophages, providing an alternative cell culture model for the study of microglia. In addition, we demonstrate that the activation of microglial cells could be modified by iron. PMID- 11424193 TI - Rumpshaker-like proteolipid protein (PLP) ratio in a mouse model with unperturbed structural and functional integrity of the myelin sheath and axons in the central nervous system. AB - The gene plp on the X chromosome encodes the isoforms proteolipid protein (PLP) and DM(20), two dominant integral membrane proteins of central nervous system (CNS) myelin. DM(20) results from the activation of the cryptic splice site in exon III of the PLP gene. We inserted a sense-orientated loxP flanked neomycin gene into intron III of the plp sequence, using homologous recombination in embryonic stem cells and generated the homozygous neoS mouse line. Unlike the previously described complete PLP/DM(20) ablation (plp(-/-)), which has been obtained by introducing a neo-gene in antisense-orientation in the same position of intron III, the plp expression surprisingly revealed reduced mRNA levels. The PLP isoform was reduced to 50%, but DM(20) expression was unaffected. This protein pattern resembles the expression profile of the PLP isoforms in the natural occurring rumpshaker mutant. Electron microscopic examination revealed a normal compaction of CNS-myelin and maintenance of axon integrity. PLP expression levels of the wt control were recovered by Cre excision of the neo-selection gene after intercrossing neoS mice and oligodendrocyte-specific Cre-mice. These data strongly hint at different functions of intron III in PLP/DM(20)-specific splicing and mRNA stability. Furthermore evidence is provided for functionally affected translation products of the PLP gene in the rumpshaker mutant, whereas no PLP-isoform occur in plp(-/-) mice generated by introducing a selectable marker into intron III in antisense orientation. PMID- 11424195 TI - Serratus posterior muscles: anatomy, clinical relevance, and function. AB - The serratus posterior superior and inferior muscles are generally considered clinically insignificant muscles that, based on attachments, probably function in respiration. Interestingly, however, there is no evidence supporting a respiratory role for these muscles. In fact, some electromyographic data refute a respiratory function for these muscles. We suggest that the serratus posterior muscles function primarily in proprioception. Further, these muscles, especially the superior, have been implicated in myofascial pain syndromes and therefore may have greater clinical relevance than commonly attributed to them. PMID- 11424194 TI - Inflammatory repertoire of Alzheimer's disease and nondemented elderly microglia in vitro. AB - We have previously developed and characterized isolated microglia and astrocyte cultures from rapid (<4 h) brain autopsies of Alzheimer's disease (AD) and nondemented elderly control (ND) patients. In the present study, we evaluate the inflammatory repertoire of AD and ND microglia cultured from white matter (corpus callosum) and gray matter (superior frontal gyrus) with respect to three major proinflammatory cytokines, three chemokines, a classical pathway complement component, a scavenger cell growth factor, and a reactive nitrogen intermediate. Significant, dose-dependent increases in the production of pro-interleukin-1beta (pro-IL-1beta), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory peptide 1alpha (MIP-1alpha), IL-8, and macrophage colony-stimulating factor (M-CSF) were observed after exposure to pre-aggregated amyloid beta peptide (1-42) (Abeta1 42). Across constitutive and Abeta-stimulated conditions, secretion of complement component C1q, a reactive nitrogen intermediate, and M-CSF was significantly higher in AD compared with ND microglia. Taken together with previous in situ hybridization findings, these results demonstrate unequivocally that elderly human microglia provide a brain endogenous source for a wide range of inflammatory mediators. PMID- 11424196 TI - Anatomical study to investigate the feasibility of pedicled nerve, free vessel gastrocnemius muscle transfer for restoration of biceps function. AB - A challenging problem is the patient with a total brachial plexus injury with nerve root avulsions. In these patients nerve repair is not possible and no local functioning muscles are available for transfer. Current techniques involve either nerve repair using donor nerves from the contralateral limb or free muscle transfer neurotized by intercostal nerves. The problem with both these techniques is that they are dependent on neural regeneration, which is imperfect. To overcome the problem we propose a technique of transferring a distant muscle whilst retaining its neural supply. Gastrocnemius is a strong muscle and one suitable for free tissue transfer. This study assessed the possibility of transferring gastrocnemius on its neural supply by determining the length of nerve available and whether it was possible to dissect the nerve to gastrocnemius from the main body of the sciatic nerve. We found that the latter was possible, and that the length of dissected nerve would allow transfer of the innervated muscle from the calf to the axilla. PMID- 11424197 TI - Pulley anatomy for the radial side of the wrist. AB - This is the first presentation in the literature of a radial tendon flexor pulley in the distal forearm adjacent to the flexor carpi radialis. The clinical significance is that in performing wrist tendon arthroplasty, this structure and, in close proximity, the sensory branch of the median nerve may be encountered. Whether cutting the pulley of the flexor carpi radialis is clinically significant in changing wrist biomechanics is unknown. PMID- 11424198 TI - Radiographic appearance of sesamoid bones in the hands and feet of Malawian subjects. AB - Our aim was to determine the prevalence of sesamoid bones in adult black Malawians from radiographs of the hands and feet. A total of 255 radiographs from 85 men and 45 women aged 16-66 years were used: 126 of the hands and 129 of the feet. All the hand films revealed one sesamoid bone at the interphalangeal joint and two at the metacarpophalangeal joint of the thumb. All the foot films showed two sesamoid bones at the metatarsophalangeal joint of the big toe. Occasional sesamoid bones were found at the metacarpophalangeal and metatarsophalangeal joints of the index finger and the small toe, respectively. Only 4.8% of the hands had sesamoid bones at the metacarpophalangeal joint of the index finger, compared with reported percentages of between 35% and 64.2% in Caucasians. The present study also records, probably for the first time in Africans, an incidence of 10.8% of sesamoid bones occurring at the metatarsophalangeal joint of the small toe, which again is at variance with the reported range of 5.5-10% in Caucasians. This study provides anatomical data that will help in the diagnosis and management of disorders of sesamoid bones which are often overlooked in Africans. PMID- 11424199 TI - Intercondylar shelf angle in adult black Malawian subjects. AB - In this study, we determined the intercondylar shelf angle of the femur in adult black Malawian subjects as measured in lateral radiographs of the knee joint. A total of 856 radiographs were used: 464 from men and 392 from women, aged 18-70 years. A goniometer measured the intercondylar shelf angle at the intersection between Blumensaat's line and a line drawn along the edge of the distal third of the femur parallel to the posterior cortex of the bone. The mean intercondylar shelf angle of 136.60 degrees +/- 5.90 degrees in Malawians of both sexes was significantly different from 142.57 degrees +/- 5.86 degrees for American subjects (P < 0.001), as given by Craig (1995, J. Forensic Sci. 40:777-782), but the level of statistical significance between black Malawian females and black American females was at P < 0.01. Using the intercondylar shelf angle, 66.67% of male Malawians were correctly identified as such and 50.00% of Malawians females as such. Eighty-three percent of males and 81.30% of females were correctly identified as blacks. This angle has been shown to exhibit sexual dimorphism in Malawian subjects. Its importance in forensic anthropology is stressed. PMID- 11424200 TI - Three-dimensional cephalometry using individual skeletal laser technology models. AB - When planning operations on the facial skull, transversal asymmetries of the maxillo-mandibular complex cannot be adequately assessed using conventional two dimensional (2D) x-ray cephalometry. On eight patients who presented with facial skull asymmetries, a three-dimensional (3D) laser technology model (LTM) using CT data was fabricated. Five sagittal plane points and six symmetry points were marked on the LTM, measured with the FlashPoint 3-D Digitizer and then geometrically converted, such that using the sagittal plane points, sella, basion, and nasion, a method could be developed that allowed the localization of each spatial point in the three symmetry planes. Thus one could quantitatively record a patient's specific facial skull asymmetry in all three planes and a 3D measurement became feasible. Based on the measurements, the asymmetry could be assessed with respect to the sagittal, vertical, and horizontal planes. With the 3-D LTM Digitizer measuring system, the surgeon now had precise numerical information regarding the symmetry ratios of the skull at his disposal, information that would have been difficult to evaluate solely using a model analysis. The results from this study show that our measuring system is applicable and useful for complex maxillofacial asymmetries. The planning of surgical interventions was optimized because precise numerical values regarding the degree of the asymmetry were available. With the 3-D LTM Digitizer measuring system, cephalometric analysis of complex asymmetries in the three spatial planes can be pragmatically supported. PMID- 11424201 TI - Upper dental prosthesis instability consecutive to the resection of a cerebral tumor. AB - A 57-year-old female patient was evaluated for an unstable maxillary dental prosthesis. The patient stated that approximately 2 weeks prior to this consultation the prosthesis was relined four times. Despite this, it often had a tendency to shift during meals. On the other hand, she is very satisfied with a complete fixed prosthesis on implants in the mandibular arch. She has been wearing this latter lower prosthesis for the same length of time as the upper prosthesis. She indicated that a similar problem existed with the former upper prosthesis, resulting in her consulting a prosthodontist. PMID- 11424202 TI - Reconstruction of the articular facets of the subtalar and talonavicular joints from volumetric magnetic resonance data. AB - Image processing methods have been used to visualize the articular surfaces of an intact foot from an MR scan and to allow the relationship between the joint surfaces to be studied. The MR protocol employed would be applicable in the living foot. Several standard image-processing methods have been used to develop protocols for manual and automated editing of a volumetric MR scan. The result is a volume-rendered foot, displaying the articular surfaces of the tarsal and metatarsal joints. The methods employed are discussed and reconstructions of the subtalar and talonavicular joints presented, thereby enabling a qualitative analysis of the facets on the talus, calcaneus and navicular. PMID- 11424203 TI - Experience of anatomy demonstrators in Tamil Nadu, India. AB - Questionnaires were sent to 118 previous holders of demonstrator posts in anatomy at the Christian Medical College, Vellore, India, in order to determine the role of these posts in the medical profession. Sixty-three (94%) of the respondents had postgraduate medical qualifications. Forty-four (66%) of the respondents were surgeons, and 23 (34%) were in non-surgical fields. Thirty-eight (57%) had jobs that involved teaching. Almost all the respondents found the teaching experience useful in their careers and especially those respondents who had qualified in surgical specialties rated their experience reviewing anatomy very highly. The usefulness of this posting is discussed. PMID- 11424204 TI - Entrapment of the lingual nerve due to an ossified pterygospinous ligament. AB - During a routine dissection course at the University of Muenster (Germany) an unusual course of the lingual nerve was found with entrapment of the nerve between a widely ossified pterygospinous ligament and the medial pterygoid muscle. Furthermore, the nerve's mobility was restricted by a more distal anastomosis with the inferior alveolar nerve. Although incomplete or complete ossification of the pterygospinous ligament is not uncommon, the combination with a medial course of the lingual nerve has not been described before. Besides practical importance for surgeons and anesthetists, the entrapment of the lingual nerve may lead to lingual numbness and pain associated with speech impairment. PMID- 11424205 TI - Rare variations of the mylohyoid muscle: case study. AB - The mylohyoid is a muscular diaphragm in the floor of the oral cavity. Its superficial and deep surfaces have important anatomical relationships. The submandibular gland is uniquely related to both surfaces at the posterior free edge of the muscle. It is here that the submandibular and sublingual tissue spaces become continuous. This case report describes an unusual range of anatomical variations of the mylohyoid muscle and reviews their clinical significance. PMID- 11424206 TI - Aberrant muscle between the temporalis and the lateral pterygoid muscles: M. pterygoideus proprius (Henle). AB - During examination of the positional relationships between the lateral pterygoid and the temporalis muscles and the innervating nerves, an aberrant muscle was observed in three of 66 head halves. The aberrant muscle originated from the medial surface of the anteromedial muscle bundle of the temporalis (Shimokawa et al. 1998, Surg. Radiol. Anat. 20:329-334) and inserted into the inferolateral surface of the lower head of the lateral pterygoid. Due to its location, origin and insertion this aberrant muscle slip is considered to correspond to the pterygoideus proprius described by Henle (1858, Handbuch der Anatomie des Menschen). Based on the innervation findings, the present aberrant muscle might be considered as a remnant muscle bundle between the anteromedial muscle bundle of the temporalis and the lateral pterygoid during differentiation of the lateral masticatory muscle anlage. PMID- 11424207 TI - Extracranial branches of the middle meningeal artery. AB - We present a case of unusual extracranial branches of the middle meningeal artery. The middle meningeal artery originated from the medial side of the maxillary artery and entered the skull through the foramen spinosum. The posterior superior alveolar artery originated from the middle meningeal artery, gave rise to the pterygoid branch, and entered the maxilla. In addition, the bifurcation of the common carotid artery was at the level of the second cervical vertebra. The embryogenesis of such rare branching pattern of the middle meningeal artery is not clear, but the anatomical consequences may have clinical implications. PMID- 11424208 TI - Foreign anatomy: a short memoir. PMID- 11424209 TI - Ran binding protein RanBP1 in zebrafish embryonic development. AB - Ran, which functions in nucleocytoplasmic transport and mitosis, binds to and is regulated in part by RanBP1. We have identified a zebrafish RanBP1 cDNA and report that it encodes for a polypeptide of 233 amino acids with considerable similarity to human and Xenopus RanBP1, despite the fact that it is 10% longer due to an extension at its carboxy terminus. RanBP1 mRNA is present as a maternal transcript and is expressed ubiquitously throughout the developing embryo. At the protein level, RanBP1 is present at all embryonic stages. Surprisingly, the ectopic overexpression of the protein had no obvious effect on embryogenesis. Attempts were also made to down-regulate RanBP1 activity by RNA interference. Injecting double-stranded RNA augmented both the mortality rate and the frequency of induced defects. Specific defects accompanied by changes in RanBP1 expression were not seen, leading us to propose that RNAi is not a reliable method for deregulating the activity of constitutively expressed genes, like RanBP1, in zebrafish. Mol. Reprod. Dev. 59:235-248, 2001. PMID- 11424210 TI - In silico mining of EST databases for novel pre-implantation embryo-specific zinc finger protein genes. AB - Progress in the understanding of early mammalian embryo development has been severely hampered by scarcity of study materials. To circumvent such a constraint, we have developed a strategy that involves a combination of in silico mining of new genes from expressed sequence tags (EST) databases and rapid determination of expression profiles of the dbEST-derived genes using a PCR-based assay and a panel of cDNA libraries derived from different developmental stages and somatic tissues. We demonstrate that in a random sample of 49 independent dbEST-derived zinc finger protein genes mined from a mouse embryonic 2-cell cDNA library, more than three-quarters of these genes are novel. Examination of characteristics of the human orthologues derived from these mouse genes reveals that many of them are associated with human malignancies. Expression studies have further led to the identification of three novel genes that are exclusively expressed in mouse embryos before or up to the 8-cell stage. Two of the genes, designated 2czf45 and 2czf48 (2czf for 2-cell zinc finger), are zinc finger protein genes coding for a RBCC protein with a RFP domain and a protein with three C2H2 fingers, respectively. The third gene, designated 2cpoz56, codes for a protein with a POZ domain that is often associated with zinc finger proteins. These three genes are candidate genes for regulatory or other functions in early embryogenesis. The strategy described in this report should generally be applicable to rapid and large-scale mining of other classes of rare genes involved in other biological and pathological processes. Mol. Reprod. Dev. 59:249 255, 2001. PMID- 11424211 TI - In vivo analysis of the regulation of the anti-Mullerian hormone, as a marker of Sertoli cell differentiation during testicular development, reveals a multi-step process. AB - Anti-Mullerian hormone (AMH) is a member of the TGF-beta family which elicits its main action during male sex differentiation. This hormone is probably the most convenient marker of Sertoli cell differentiation and maturation throughout testicular development. Studying AMH gene regulation may thus be one way of identifying effectors of Sertoli cell differentiation. To this end we first tried to locate and then to characterise DNA elements responsible for in vivo transcriptional control of AMH expression. We obtained transgenic mice expressing a reporter gene (LacZ), under control of various putative AMH regulatory sequences. Analysis of transgenic animals revealed that activation of the AMH gene probably requires a two-step regulatory process. The first step corresponds to the initial activation of the AMH gene occurring at around 12.0 dpc. It requires the presence of regulatory DNA encompassed within a maximum of 370 bp upstream of the translation start site of the gene, delimited by the presence of an upstream housekeeping gene (SAP-62). Following this initial transient phase, a second phase seems to account for the persistence of AMH gene expression until the onset of puberty. As the 370 bp regulatory region is not sufficient on its own to allow the triggering of this second phase, it seems possible that additional control elements are required for normal AMH expression throughout testicular development. The complete array of regulatory elements remains to be located. Mol. Reprod. Dev. 59:256-264, 2001. PMID- 11424212 TI - Aromatase plays a key role during normal and temperature-induced sex differentiation of tilapia Oreochromis niloticus. AB - In the tilapia Oreochromis niloticus, sex is determined genetically (GSD), by temperature (TSD) or by temperature/genotype interactions. Functional masculinization can be achieved by applying high rearing temperatures during a critical period of sex differentiation. Estrogens play an important role in female differentiation of non-mammalian vertebrates. The involvement of aromatase, was assessed during the natural (genetic all-females and all-males at 27 degrees C) and temperature-induced sex differentiation of tilapia (genetic all females at 35 degrees C). Gonads were dissected between 486--702 degree x days. Aromatase gene expression was analyzed by virtual northern and semi-quantitative RT-PCR revealing a strong expression during normal ovarian differentiation concomitant with high levels (465 +/- 137 fg/g) of oestradiol-17 beta (E2-17 beta). This was encountered in gonads after the onset of ovarian differentiation (proliferation of both stromal and germ cells prior to ovarian meiosis). Genetic males exhibited lower levels of aromatase gene expression and E2-17 beta quantities (71 +/- 23 fg/ g). Aromatase enzyme activity in fry heads established a sexual dimorphism in the brain, with high activity in females (377.9 pmol/head/hr) and low activity in males (221.53 pmol/head/hr). Temperature induced the masculinization of genetic females to a different degree in each progeny, but in all cases repression of aromatase expression was encountered. Genetic males at 35 degrees C also exhibited a repression of aromatase expression. Aromatase brain activity decreased by nearly three-fold in the temperature-masculinized females with also a reduction observed in genetic males at 35 degrees C. This suggests that aromatase repression is required in the gonad (and perhaps in the brain) in order to drive differentiation towards testis development. Mol. Reprod. Dev. 59:265-276, 2001. PMID- 11424213 TI - Nopp 140 involvement in nucleologenesis of mouse preimplantation embryos. AB - As it was shown earlier, resumption of rRNA transcription in early mouse embryo is localized in the peripheral region of nucleolus precursor body/NPB/during the two-cell stage. Recently, nucleolar phosphoprotein Nopp140 was presented to shuttle between the nucleolus and cytoplasm as chaperone of snoRNPs. Nopp140 interacts with RNA polymerase I in nucleolus and also accumulates in CBs, suggesting a pathway between the two organelles. The aim of the study was to describe the changing location of Nopp140 during the first cleavage stages of mouse embryos and its re-location after inhibition of rRNA synthesis with actinomycin D. Light microscope immunocytochemical staining showed Nopp140 in the periphery of NPBs before activation of rDNA transcription and in addition confirmed its localization in CBs. Immunolabelling with antibodies against RNA Pol I and UBF gave co-localization of these proteins, implicating that Nopp140 may actively participate to rDNA transcription. We suggest that fundamental differences in molecular organization of rDNA synthesis and postranscriptional processes between cycling somatic and pre-implantation embryonic cells may be in selective transport of transcription and/or processing-complexes of proteins to the nucleolar organizer regions (NOR). Mol. Reprod. Dev. 59:277-284, 2001. PMID- 11424214 TI - Heat shock proteins Hsp70-1 and Hsp70-3 Are necessary and sufficient to prevent arsenite-induced dysmorphology in mouse embryos. AB - Heat Shock Proteins (HSPs) represent a variety of protein families that are induced by stressors such as heat and toxicants, and the induction of HSPs in the organogenesis stage rodent embryo is well established. It has been proposed that thermotolerance and chemotolerance result from expression of the HSPs. However, whether these proteins function to prevent dysmorphogenesis and which family members serve this function are unknown. Therefore, we evaluated the specific ability of stress-inducible Hsp70-1 and Hsp70-3 to prevent arsenite-induced dysmorphology in the cultured mouse embryo using gain- and loss-of-function models. Loss of HSP function was accomplished by injecting antisense oligonucleotides directed against hsp70-1 and hsp 70-3 mRNAs into the amniotic cavity of cultured Day 9 mouse embryos. Suppression of hsp70-1 and hsp70-3 expression resulted in an up to six-fold increase in the incidence of arsenite induced neural tube defects. Gain of HSP function was accomplished by microinjecting a transgene with a constitutive promotor driving expression of the hsp70-1 coding region, and resulted in a decreased incidence of arsenite-induced neural tube defects. These results indicate that Hsp70-1 and Hsp70-3 are both necessary and sufficient for preventing arsenite-induced dysmorphology in early somite staged mouse embryos. Mol. Reprod. Dev. 59:285-293, 2001. PMID- 11424215 TI - Isolation and characterization of sea urchin egg lipid rafts and their possible function during fertilization. AB - Specialized membrane microdomains called rafts are thought to play a role in many types of cell-cell interactions and signaling. We have investigated the possibility that sea urchin eggs contain these specialized membrane microdomains and if they play a role in signal transduction at fertilization. A low density, TX-100 insoluble membrane fraction, typical of lipid rafts, was isolated by equilibrium gradient centrifugation. This raft fraction contained proteins distinct from cytoskeletal complexes. The fraction was enriched in tyrosine phosphorylated proteins and contained two proteins known to be involved in signaling during egg activation (an egg Src-type kinase and PLC gamma). This fraction was further characterized as a prototypical raft fraction by the release of proteins in response to in vitro treatment of the rafts with the cholesterol binding drug, methyl-beta-cyclodextrin (M beta CD). Furthermore, treatment of eggs with M beta CD inhibited fertilization, suggesting that egg lipid rafts play a physiological role in fertilization. Mol. Reprod. Dev. 59:294-305, 2001. PMID- 11424216 TI - Lipid dynamics in the plasma membrane of ram and bull spermatozoa after washing and exposure to macromolecules BSA and PVP. AB - Seminal plasma proteins and macromolecules in the external medium have a major influence on the functionality of sperm plasma membranes. In this investigation we have examined their effects on lipid diffusion in the surface membrane of ram and bull spermatozoa as measured by fluorescence recovery after photobleaching (FRAP). Results show that progressive removal of seminal plasma from ram spermatozoa by repeated centrifugation and resuspension in media +/- 4% bovine serum albumin (BSA) or 0.4% polyvinlypyrrolidone (PVP) causes a reduction in lipid diffusion in all regions of the membrane. By contrast, bull sperm membranes respond with an increase in diffusion in all regions. Repeated washing of bull spermatozoa whose membranes were previously immobile (i.e., showed no recovery after FRAP) restored lipid diffusion suggesting an inhibitory effect of seminal plasma proteins. Further analysis by atomic force microscopy revealed a close association between BSA and the plasma membrane. It is concluded that diffusion of lipids in the plasma membrane of ejaculated ram and bull spermatozoa is influenced by seminal plasma proteins and the composition of the suspending medium. Mol. Reprod. Dev. 59:306-313, 2001. PMID- 11424217 TI - Nuclear matrix in developing rat spermatogenic cells. AB - The nonchromatin structure or nuclear matrix in developing spermatogenic cells of the rat was studied using a biochemical fractionation in concert with resinless section electron microscopy. Observations demonstrated that the nuclear matrix of spermatogenic cells consisted of a three-dimensional network of filaments of variable thicknesses. In spermatogonia and spermatocytes the nuclear matrix consisted of relatively thin filaments, while that of round spermatids consisted of a thicker interconnecting network of filament. In elongating spermatids, the interior of the nuclear matrix consisted of a network of dense filaments bounded by a peripheral lamina. The protein composition of the nuclear matrix in spermatogenic cells was examined by high-resolution two-dimensional gel electrophoresis and correlated with morphological changes characteristic of each stage. The results showed that the proteins of nuclear matrix changed in a cell stage-specific manner. These stage-specific changes corresponded to the major transitions of chromatin structure and function during spermatogenesis. Furthermore, immunocytochemical and immunoblotting analysis of DNA topoisomerase II (topo II) revealed that this enzyme exhibited stage-specific variations and was associated with the nuclear matrix. These results suggest that the nuclear matrix in spermatogenic cells may be involved in mediating DNA modifications and maintaining nuclear organization during spermatogenesis. Mol. Reprod. Dev. 59:314 321, 2001. PMID- 11424218 TI - VLA-6 integrin distribution and calcium signalling in capacitated boar sperm. AB - Previous investigations showed that VLA-6 integrin present on boar sperm membrane can induce acrosome reaction upon exposure to laminin accumulated in expanded cumuli (Mattioli et al., 1998. To further investigate this novel sperm egg recognition system, the authors studied the distribution of VLA-6 integrin on the membrane of boar sperm throughout capacitation and following acrosome reaction, and analyzed intracellular Ca(2+) changes occurring in spermatozoa exposed to laminin. Immunofluorescent localisation of VLA-6 revealed a low proportion (nearly 22%) of positive cells in freshly ejaculated sperm, with integrin mainly concentrated in clustered spots. After 3 hr incubation most of the spermatozoa showed integrin molecules on the membrane, with three different labeling patterns: fluorescence localised on the edge of the acrosome (58.2 +/- 14.2% of the cells); fluorescence uniformly spread over the whole sperm head (5.0 +/- 1.9%) and finally fluorescence concentrated in clustered spots (7.6 +/- 5.6%), as recorded in freshly ejaculated sperm. Twenty-nine percent of cells did not show any distinct fluorescence. Following acrosome reaction sperm with fluorescence on the acrosomal region virtually disappeared and the proportion of unstained cells rose from 29.2 +/- 9.2 to 69.0 +/- 10.1%. Electron microscopy demonstrated that VLA-6 integrin was exclusively located on the sperm membrane of intact spermatozoa. Confocal analysis showed that laminin triggers distinct Ca(2+) raises, and that sperm exposed and kept in the presence of laminin fully retained their ability to rise intracellular Ca(2+) in response to zona pellucida proteins. These data indicate that boar sperm accumulate VLA-6 integrin on the membrane and concentrate it on the acrosomal region as capacitation progresses. Probably due to this compartmentalisation, sperm exposed to laminin experience a Ca(2+) raise that originates in the anterior sperm head where it is more adequate for the induction of acrosome reaction. Mol. Reprod. Dev. 59:322-329, 2001. PMID- 11424219 TI - Cysteine, glutathione, and Percoll treatments improve porcine oocyte maturation and fertilization in vitro. AB - Several factors have been examined to improve in vitro fertilization and development of porcine oocytes. Cysteine is known to be beneficial for oocyte maturation and male pronuclear formation in pigs and glutathione is known to help prevent membrane disruption of sperm in other species, including human. It has also been reported that the presence of cumulus cells influences the outcome of in vitro fertilization in cattle. The objectives of the present study were to investigate the effects of several factors involved in porcine in vitro maturation (IVM) and fertilization (IVF) procedures on oocyte embryogenic competence. The following factors were examined: the effects of different concentrations (0, 0.285, 0.57, 1.14, 2.28 microM) and exposure duration (22 and 44 hr) of cysteine during IVM, glutathione inclusion and of cumulus presence during IVF, and the use of gradient Percoll (45%/90%) during sperm preparation. The presence of cysteine in maturation medium improved blastocyst development significantly regardless of the duration of exposure when compared to the control (11--16% vs. 4%, P < 0.01). However, no dose-responsive effect was observed at the concentrations tested. The use of gradient Percoll during sperm preparation significantly improved cleavage (85% vs. 57%, P < 0.01) and blastocyst development (24% vs. 6%, P < 0.01) over conventional sperm preparation. Significant improvement was also achieved by the addition of glutathione to Percoll gradient (30% vs. 20%, P < 0.05). In conclusion, cysteine and glutathione as well as Percoll and cumulus were beneficial to embryogenic competence of porcine oocytes in this study. Mol. Reprod. Dev. 59:330-335, 2001. PMID- 11424220 TI - Estrogen receptor- and aromatase-deficient mice provide insight into the roles of estrogen within the ovary and uterus. AB - Estrogen receptor (ER)- (alpha, beta, and both alpha and beta) and aromatase (Ar) knockout (KO) mice have been created to assess the biological effects of estrogens. This review article discusses the ovarian and uterine phenotypes of these mice. The data obtained have confirmed some older inferences about how the steroid acts, but have also revealed some unexpected aspects of estrogen action. Mol. Reprod. Dev. 59:336-346, 2001. PMID- 11424221 TI - Real-time and quantitative PCR: applications to mechanism-based toxicology. AB - There is increasing awareness that quantitative analysis of changes in molecular targets plays a key role in addressing scientific questions in molecular toxicology, molecular epidemiology, and human risk assessment. One of the emerging technologies that is being used to analyze these molecular targets is real-time and quantitative (RTAQ) polymerase chain reaction (PCR). The aim of this review is to provide the reader with an overview of this technology and to highlight specific applications of this technology to some key areas of molecular toxicology. PMID- 11424222 TI - Regulation of the GST Mu-1 isoenzyme in Y1 cells by adrenocorticotropic hormone is primarily transcriptional. AB - In earlier experiments, adrenocorticotropic hormone (ACTH) was shown to decrease the level of glutathione transferase M1 (murine glutathione transferase mu-1) (mGSTM1), as well as of the corresponding mRNA, in a murine adrenocortical cell line. In the present study, the effect of ACTH on mGSTM1 gene transcription was examined using two techniques. First, a cDNA that coded for the mGSTM1 subunit but lacked the corresponding promoter sequences was transfected into the adrenocortical cell line, and the effect of ACTH on the level of the corresponding transcript was compared to that of endogenous mGSTM1 mRNA. The other technique used was nuclear run-on transcription, where the rate of transcription of endogenous mGSTM1 mRNA in ACTH-treated cells was compared to that in untreated control cells. These experimental approaches indicated that the rate of transcription of the mGSTM1 gene is regulated by ACTH in adrenocortical cells. PMID- 11424223 TI - Use of real-time gene-specific polymerase chain reaction to measure RNA expression of three family members of rat cytochrome P450 4A. AB - Exposure of rats to peroxisome proliferators induces members of the cytochrome P450 4A (CYP4A) family. In rats, the CYP4A family consists of four related genes, CYP4A1, CYP4A2, CYP4A3, and CYP4A8. We are specifically interested in examining CYP4A1, CYP4A2, and CYP4A3, each of which is expressed in a tissue-dependent and sex-dependent manner. While CYP4A1 is sufficiently different from the other two members to enable relatively easy specific quantitation, the close similarity between CYP4A2 and CYP4A3 makes quantitative discrimination difficult. We have combined a fluorescent real-time PCR assay (TaqMan) with the sequence-specific mismatch amplification mutation assay (MAMA) to allow us to carry out specific quantitation of all three members of this family. The assay is designed such that a single fluorescent TaqMan(R) probe binds to all three gene products, while specificity is conferred by sequence-specific primers. This specific MAMA technique takes advantage of the ability of Taq polymerase to distinguish between the two cDNAs based on mismatches at the 3' end of a PCR primer. In the 84-base PCR product used for this assay, there is only a single-base difference between CYP4A2 and CYP4A3. Despite this similarity, there is at least a 1000-fold discrimination between the two sequences, using CYP4A2 or CYP4A3 specific standards. Analysis of rat liver RNA from both sexes demonstrates that this discrimination is also achieved in complex RNA mixtures. This technique should be broadly applicable to other areas of research such as allelic discrimination, detecting mutational hotspots in tumors, and discrimination among closely related members of other gene families. PMID- 11424224 TI - Effects of quercetin on antioxidant defense in streptozotocin-induced diabetic rats. AB - In light of evidence that some complications of diabetes mellitus may be caused or exacerbated by oxidative damage, we investigated the effects of subacute treatment with the antioxidant quercetin on tissue antioxidant defense systems in streptozotocin-induced diabetic Sprague-Dawley rats (30 days after streptozotocin induction). Quercetin, 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H-1-benzopyran-4 one, was administered at a dose of 10mg/kg/day, ip for 14 days, after which liver, kidney, brain, and heart were assayed for degree of lipid peroxidation, reduced and oxidized glutathione content, and activities of the free-radical detoxifying enzymes catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase. Treatment of normal rats with quercetin increased serum AST and increased hepatic concentration of oxidized glutathione. All tissues from diabetic animals exhibited disturbances in antioxidant defense when compared with normal controls. Quercetin treatment of diabetic rats reversed only the diabetic effects on brain oxidized glutathione concentration and on hepatic glutathione peroxidase activity. By contrast, a 20% increase in hepatic lipid peroxidation, a 40% decline in hepatic glutathione concentration, an increase in renal (23%) and cardiac (40%) glutathione peroxidase activities, and a 65% increase in cardiac catalase activity reflect intensified diabetic effects after treatment with quercetin. These results call into question the ability of therapy with the antioxidant quercetin to reverse diabetic oxidative stress in an overall sense. PMID- 11424225 TI - Changes in the levels of glutathione after cellular and cutaneous damage induced by squalene monohydroperoxide. AB - Squalene monohydroperoxide (Sq-OOH), the initial product of ultraviolet peroxidated squalene, was used to investigate the effect of peroxidative challenge upon the glutathione contents in rabbit ear skin and primary-cultured fibroblasts derived from rabbit ear skin. The cellular reduced glutathione (GSH) contents decreased during 30-minute incubations in vitro with Sq-OOH, and oxidized glutathione (GSSG) was formed concomitantly, indicating that Sq-OOH had a potential for GSH-depleting activity in vitro. When Sq-OOH was applied topically to the skin in vivo, only GSSG contents increased significantly within 30 minutes. Moreover, pretreatment with the GSH depletors, DL-buthionine sulfoximine (BSO) and diethyl maleate (DEM), could potentiate the cytotoxicity and comedogenicity induced by Sq-OOH. These findings suggest that the endogenous antioxidant, glutathione, is quite sensitive to Sq-OOH and may be an important material for protecting cells and/or tissues against the oxidative stress induced by Sq-OOH treatment. PMID- 11424226 TI - Effects of isoeugenol on oxidative stress pathways in normal and streptozotocin induced diabetic rats. AB - Because some complications of diabetes mellitus may result from oxidative damage, we investigated the effects of subacute treatment (10mg/kg/day, intraperitoneal [ip], for 14 days) with the antioxidant isoeugenol on the oxidant defense system in normal and 30-day streptozotocin-induced diabetic Sprague-Dawley rats. Liver, kidney, brain, and heart were assayed for degree of lipid peroxidation, reduced and oxidized glutathione content, and activities of the free radical-detoxifying enzymes catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase. All tissues from diabetic animals exhibited disturbances in antioxidant defense when compared with normal controls. Treatment with isoeugenol reversed diabetic effects on hepatic glutathione peroxidase activity and on oxidized glutathione concentration in brain. Treatment with the lipophilic compound isoeugenol also decreased lipid peroxidation in both liver and heart of normal animals and decreased hepatic oxidized glutathione content in both normal and diabetic rats. Some effects of isoeugenol treatment, such as decreased activity of hepatic superoxide dismutase and glutathione reductase in diabetic rats, were unrelated to the oxidative effects of diabetes. In heart of diabetic animals, isoeugenol treatment resulted in an exacerbation of already elevated activities of catalase. These results indicate that isoeugenol therapy may not reverse diabetic oxidative stress in an overall sense. PMID- 11424227 TI - Purification and characterization of rat hepatic microsomal low molecular weight fatty acid ethyl ester synthase and its relationship to carboxylesterases. AB - We reported purification of a high molecular weight (HMW) (ca. 180 kD) and a low molecular weight (LMW) (ca. 60 kD) protein fractions from digitonized rat liver microsomes using ammonium sulfate precipitation followed by ion exchange and gel filtration column chromatography. Both fractions expressed fatty acid ethyl ester (FAEE) synthase as well as p-nitrophenyl acetate (PNPA)-hydrolyzing (esterase) activities. The HMW fraction was found to be a trimer with subunit molecular weight ca. 60 kD and structurally and functionally similar to rat hepatic microsomal carboxylesterase (CE, pI 6.1) and adipose tissue FAEE synthase. In this article, we report further purification and characterization of the LMW (minor) fraction expressing FAEE synthase activity and its structural and functional relationship to hepatic microsomal CEs. Using isoelectric focusing (IEF) followed by gel filtration-high-performance liquid chromatography (GF HPLC), five proteins were purified, which expressed FAEE synthase as well as PNPA hydrolyzing activity. The isoelectric point values of 6.5, 5.8, 5.6, 5.3, and 5.0 were found for the purified LMW proteins by IEF and each showed a peak corresponding to ca. 60 kD molecular weight by GF-HPLC, which expressed FAEE synthase as well as PNPA-hydrolyzing activity. Sodium dodecyl sulfate polyacrylamide gel elecrophoresis (SDS-PAGE) analysis of the GF-HPLC purified LMW proteins revealed that these proteins are monomers (ca. 60 kD). All the purified LMW proteins cross-reacted with antibodies to rat adipose tissue FAEE synthase. Coelution of PNPA-hydrolyzing and FAEE synthase activity at each step of purification and cross-reactivity with rat adipose tissue FAEE synthase antibodies suggest that the purified proteins are related to various hepatic microsomal CEs. This conclusion is further supported by the homology of N terminal amino acid sequence of the purified LMW proteins to various hepatic microsomal CEs and protease precursors. Therefore, LMW FAEE synthase activity most probably is expressed by various isozymes of hepatic microsomal CEs, which are also involved in the biotransformation of xenobiotic alcohols and amines. PMID- 11424229 TI - The importance of first-phase insulin secretion: implications for the therapy of type 2 diabetes mellitus. AB - Type 2 diabetes is a heterogeneous disorder characterized by defects in insulin secretion and action. Insulin resistance is a key feature of type 2 diabetes. However, insulin resistance alone does not appear to be sufficient to cause diabetes. Longitudinal studies have shown that the development of overt hyperglycemia is associated with a decline in beta-cell secretion. In patients with impaired glucose tolerance or in the early stages of type 2 diabetes, first phase insulin release is almost invariably lost despite the enhancement of second phase secretion. Both animal and human studies support the critical physiologic role of the first-phase of insulin secretion in the maintenance of postmeal glucose homeostasis. This effect is primarily mediated at the level of the liver, allowing prompt inhibition of endogenous glucose production (EGP) and thereby restraining the mealtime rise in plasma glucose. In type 2 diabetes, the loss of the early surge of insulin release is a precocious and quite common defect that plays a pathogenic role in postmeal hyperglycemia and one that may require specific therapeutic intervention. This becomes even more apparent if the negative impact of prandial glucose spikes is taken into consideration. Epidemiological evidence exists to indicate that 2-h postload plasma glucose levels are strongly associated with all-cause and cardiovascular mortality relative risk. Indeed the acute elevation of plasma glucose concentration triggers an array of tissue responses that may contribute to the development of diabetic complications. Considering that type 2 diabetes begins with meal-related hyperglycemia in many patients, it becomes apparent that normalization of postmeal plasma glucose levels should be the target for rational therapy and the goal in the early stages of the disease. If a primary goal of diabetes therapy is control of postmeal glucose excursion, then the regulation of glucose absorption from the gut and entry into the circulation is an important mechanism to consider. The restoration of the rapid increase in plasma insulin concentration may be quite an efficient therapeutic approach. PMID- 11424230 TI - Coincident diabetes mellitus and primary hyperparathyroidism. AB - The prevalence of diabetes mellitus in primary hyperparathyroidism is approximately 8% and that of primary hyperparathyroidism in diabetic patients is approximately 1%. Both values are about three-fold higher than the respective expected prevalences in general populations. Patients with both disorders are over 40 years of age and 80% are female; 22% have type 1 and 78% type 2 diabetes. Primary hyperparathyroidism presents first in approximately 20% of patients, and diabetes mellitus in 40%; both disorders present together, or within 1 year, in 40%. Approximately 40% of patients with primary hyperparathyroidism have impaired glucose tolerance. Insulin resistance is present in hyperparathyroidism and probably arises from a raised intracellular free calcium concentration which, by decreasing normal insulin-stimulated glucose transport, increases the requirement for insulin: if this insulin resistance progresses, impaired glucose tolerance and diabetes mellitus would result. Parathyroidectomy has been followed by regression of diabetes and of impaired glucose tolerance in some but not all patients. Early diagnosis of the second disorder is clinically desirable when one disorder is present. Hyperparathyroid patients should therefore be screened for impaired glucose tolerance and diabetes annually, and pre-operatively. Diabetic patients should be checked for hypercalcaemia at appropriate intervals; although only 1% of them may have hyperparathyroidism, this disorder if untreated is associated with hypertension, to which diabetic patients are already prone. PMID- 11424231 TI - Clinical importance of insulin secretion and its interaction with insulin resistance in the treatment of type 2 diabetes mellitus and its complications. AB - Type 2 diabetes primarily develops from pathogenic defects in the mechanisms of insulin secretion and hepatic and peripheral insulin action. The consequent disruption of normal glucose metabolism involves a number of organ systems and is ultimately manifested in fasting and daytime hyperglycemia. Chronically elevated blood glucose concentrations determine the progression of the disease by further exacerbating insulin resistance and causing beta-cell exhaustion in addition to decreasing their responsiveness to glucose. The beta-cell secretory dysfunction is characterized by the lack of the early phase of glucose-induced insulin secretion and the insufficient and delayed late phase of secretion. Glycemic levels in patients with type 2 diabetes are directly related to the risk of developing microvascular and macrovascular complications, the main cause of the morbidity and mortality associated with this disease. The goal of treatment is to decrease the risk and delay the progression of these complications by improving glycemic control. Current oral antidiabetic agents, used as monotherapy or in combination, include traditional insulin secretagogues, insulin sensitizers and inhibitors of carbohydrate absorption. A greater understanding of the pathophysiology of type 2 diabetes and recent findings on the significance of meal-related glycemia to overall glycemic control are expanding the therapeutic options for treating this disease. PMID- 11424232 TI - The role of oxidative stress in the onset and progression of diabetes and its complications: a summary of a Congress Series sponsored by UNESCO-MCBN, the American Diabetes Association and the German Diabetes Society. AB - This review summarises the results and discussions of an UNESCO-MCBN supported symposium on oxidative stress and its role in the onset and progression of diabetes. There is convincing experimental and clinical evidence that the generation of reactive oxygen species (ROI) is increased in both types of diabetes and that the onset of diabetes is closely associated with oxidative stress. Nevertheless there is controversy about which markers of oxidative stress are most reliable and suitable for clinical practice. There are various mechanisms that contribute to the formation of ROI. It is generally accepted that vascular cells and especially the endothelium become one major source of ROI. An important role of oxidative stress for the development of vascular and neurological complications is suggested by experimental and clinical studies. The precise mechanisms by which oxidative stress may accelerate the development of complications in diabetes are only partly known. There is however evidence for a role of protein kinase C, advanced glycation end products (AGE) and activation of transcription factors such as NF kappa B, but the exact signalling pathways and the interactions with ROI remain a matter of discussion. Additionally, results of very recent studies suggest a role for ROI in the development of insulin resistance. ROI interfere with insulin signalling at various levels and are able to inhibit the translocation of GLUT4 in the plasma membrane. Evidence for a protective effect of antioxidants has been presented in experimental studies, but conclusive evidence from patient studies is missing. Large-scale clinical trials such as the DCCT Study or the UKPDS Study are needed to evaluate the long-term effects of antioxidants in diabetic patients and their potential to reduce the medical and socio-economic burden of diabetes and its complications. PMID- 11424233 TI - Genomic variation in pancreatic ion channel genes in Japanese type 2 diabetic patients. AB - BACKGROUND: Many genetic diseases are caused by mutations in ion channel genes. Because type 2 diabetes is characterized by pancreatic beta-cell insensitivity to glucose, the genes responsible for glucose metabolism and calcium signaling in pancreatic beta-cells are candidate type 2 diabetes susceptibility genes. METHODS: We have examined genomic variations in two ion channel genes relevant to the molecular pathology of diabetes mellitus, the Kir6.2 subunit of the ATP sensitive potassium channel gene and alpha(1D) subunit of the voltage-dependent calcium channel (VDCC) gene among Japanese type 2 diabetic patients. RESULTS: There are two alleles in the Kir6.2 gene: EI, glutamic acid at codon 23 and isoleucine at codon 337 and KV, lysine at codon 23 and valine at codon 337. The allelic frequencies of these polymorphisms are similar in type 2 diabetic patients and normal subjects. We also detected trinucleotide repeat polymorphisms in the amino terminus and the carboxyl terminal region of the alpha(1D) gene. Expansion of the ATG trinucleotide repeat from seven to eight was detected only in type 2 diabetic patients, but the frequency was low and was similar in type 2 diabetic patients and normal subjects. CONCLUSIONS: Although variations of the Kir6.2 and alpha(1D) genes are not associated with the development of common type 2 diabetes, further studies may determine the role of these genomic variations, especially those in the alpha(1D) VDCC gene, in the pathogenesis of certain subsets of type 2 diabetes, or as a co-factor in the polygenic disorder generally. PMID- 11424234 TI - Effect of D-glucose on nitric oxide release from glomerular endothelial cells. AB - BACKGROUND: Altered glomerular production of nitric oxide (NO) may be involved in hyperfiltration in early diabetic nephropathy. However little is known as to the role of glomerular endothelial cells (GECs) in diabetic hyperfiltration and their ability to release NO in response to hyperglycemia. METHODS: Using an established cell line, we directly monitored NO release from GECs in response to various concentrations of D-glucose, D-mannitol, and L-arginine, an NO synthase (NOS) agonist. L-Arginine-induced NO release was examined in the cells pretreated for different periods up to 24 h with 10 or 30 mM D-glucose. We also measured serially the accumulation of nitrite, the stable metabolite of NO, produced by the cells incubated for up to 24 h under 10 or 30 mM D-glucose conditions in the presence or absence of the NOS inhibitor, L-NAME. RESULTS: Direct measurement of NO demonstrated that D-glucose, but not D-mannitol, stimulation resulted in a rapid and dose-dependent increase in NO release by the cells. However, L-arginine induced NO release was attenuated significantly in the cells preincubated for more than 12 h with 30 mM D-glucose compared to 10 mM D-glucose. The L-NAME inhibitable production of nitrite in the media was significantly increased 1.5- 2.0-fold until 6 h after incubation with 30 mM D-glucose compared to 10 mM D glucose. CONCLUSIONS: We conclude that D-glucose, but not D-mannitol, produces a rapid and dose-dependent increase in NO release, whereas exposure to high D glucose for more than 12 h may blunt NOS activity and/or NO stability in the GECs. These observations may therefore be important for glomerular endothelial dysfunction induced by hyperglycemia that is still tentative and may have a role in diabetic nephropathy. PMID- 11424236 TI - Current literature in diabetes. PMID- 11424235 TI - Brain and liver mitochondria isolated from diabetic Goto-Kakizaki rats show different susceptibility to induced oxidative stress. AB - BACKGROUND: Increased oxidative stress and changes in antioxidant capacity observed in both clinical and experimental diabetes mellitus have been implicated in the etiology of chronic diabetic complications. Many authors have shown that hyperglycemia leads to an increase in lipid peroxidation in diabetic patients and animals reflecting a rise in reactive oxygen species production. The aim of the study was to compare the susceptibility of mitochondria from brain and liver of Goto-Kakizaki (12-month-old diabetic) rats (GK rats), a model of non-insulin dependent diabetes mellitus, to oxidative stress and antioxidant defenses. METHODS: Brain and liver mitochondrial preparations were obtained by differential centrifugation. Oxidative damage injury was induced in vitro by the oxidant pair ADP/Fe(2+) and the extent of membrane oxidation was assessed by oxygen consumption, malondialdehyde (MDA) and thiobarbituric acid reactive substances (TBARS) formation. Coenzyme Q and alpha-tocopherol contents were measured by high performance liquid chromatography (HPLC). RESULTS: Brain mitochondria isolated from 12-month-old control rats displayed a higher susceptibility to lipid peroxidation, as assessed by oxygen consumption and formation of MDA and TBARS, compared to liver mitochondria. In GK rats, mitochondria isolated from brain were more susceptible to in vitro oxidative damage than brain mitochondria from normal rats. In contrast, liver mitochondria from diabetic rats were less susceptible to oxidative damage than mitochondria from normal rats. This decreased susceptibility was inversely related to their alpha-tocopherol and coenzyme Q (CoQ) content. CONCLUSIONS: The present results indicate that the diabetic state can result in an elevation of both alpha-tocopherol and CoQ content in liver, which may be involved in the elimination of mitochondrially generated reactive oxygen species. The difference in the antioxidant defense mechanisms in the brain and liver mitochondrial preparations of moderately hyperglycemic diabetic GK rats may correspond to a different adaptive response of the cells to the increased oxidative damage in diabetes. PMID- 11424237 TI - Abortion and politics in Mexico: 'context is all'. AB - A strong collective pro-choice mentality was recently manifested in Mexico when a legislative initiative to revoke the legal right of rape survivor to abortion in the state of Guanajuato awakened national indignation. Pro-choice values were expressed in public opinion with such force that it sparked off the passage of liberalising law reforms in Mexico City and the state of Morelos. In this paper we trace the development of these manifestations of pro-choice views, beginning with the Democratic Revolution Party's (PRD) refusal in 1999 to modify abortion legislation within the context of penal code reform, and moving through the events surrounding the Guanajuato reform, and the pro-choice response of Mexico City and Morelos legislators. This analysis allows us to recognise the emergence of a pro-choice consciousness and to understand that, when it comes to abortion, 'context is all'. PMID- 11424238 TI - The role of advocacy in implementing the South African abortion law. AB - In order to translate the abortion law in South Africa into services that ensure equity of access and women's right to control their bodies, interventions are needed to change judgemental views on abortion. This paper describes formative research conducted in the Northern Cape Province among 436 community members, 29 women seeking an abortion and 80 health service providers, to develop appropriate interventions to these ends. Based on the findings, two interventions were developed. These interventions appeared to substantially influence personal views by getting people to make a connection between the need for abortion services and the circumstances in which unwanted pregnancies occur. There was a shift towards greater support for women's right to choose in relation to abortion among women community members, though not among men, who resisted this right for married women. Amongst providers (almost all women) there was an increase in willingness to support service provision and to support staff working in abortion services. These tools could be used in sexuality education in schools and in nurse midwifery/medical training, to complement current advocacy initiatives taking place at policy and programme levels in South Africa, to help to reduce the public health problem of unsafe abortion. PMID- 11424239 TI - Integration of STI services into FP/MCH services: health service and social contexts in rural Ghana. AB - Ghana, like many countries, has taken up the call to integrate STI management with MCH/FP services. Since 1994 a range of policies has been developed on safe motherhood and family planning, as well as syndromic STI management guidelines for 'mid-level' staff and an all-encompassing 'reproductive health' policy. The success of these policy initiatives depends to a large extent on the processes and contexts of implementation, yet analysis of this has rarely been undertaken. This paper analyses the rural context of policy implementation in the rural Upper East Region of northern Ghana and suggests that a 'blanket' policy to integrate STI and FP/MCH services may be inappropriate in particular contexts. It illustrates how the implementation of health policies is influenced--and often impeded--not only by local service contexts, economic and epidemiological factors but also by culturally defined social attitudes and behaviours. These can influence not only whether a policy is accepted by a population but also how it is implemented by health staff. Future reproductive health policy needs to address these issues in order to develop goals which can realistically be implemented in specific settings. Enhancing at district level the voice of nurses working at community level and promoting collaborative, culturally-specific and community-based initiatives could facilitate this. PMID- 11424240 TI - The Copper-T 380A IUD: a ten-year alternative to female sterilisation in India. AB - High unmet need for limiting contraception persists in most states of India despite wide access to sterilisation. Qualitative evidence from a rural community in which child mortality is high and women's autonomy is low suggests that women may seek reversibility in a contraceptive even if they have finished childbearing. This paper describes the introduction of the Copper-T 380A--a contraceptive with an effective life span of ten years--as an alternative to female sterilisation in a rural area of the state of Rajasthan, in a clinic linked to an outreach programme. The intervention addressed women's apprehensions, ensured service standards and guaranteed women's right to have the Copper-T removed at will. Data on 216 insertions over 34 months revealed a preference for the Copper-T 380A among older women and women who had achieved desired family size, especially among tribal women. More than a quarter of the 30 removals in that period were for non-medical reasons, such as family opposition, child death or remarriage. As a long-term but reversible option, the Copper-T 380A allows women room to change their minds in relation to future childbearing until they have reached menopause. Including this option in family planning services can help to meet a portion of the unmet need for contraception among women not willing to choose sterilisation, while reducing dependence on doctors and expensive equipment. PMID- 11424241 TI - Unmet needs, beliefs and treatment-seeking for infertility among migrant Ghanaian women in the Netherlands. AB - Studies on infertility in the Netherlands have little information on migrant Ghanaian women, even though Ghanaians are the third largest migrant group in Amsterdam. An exploratory study on the unmet needs, attitudes, and beliefs of migrant Ghanaian women with infertility problems living in the Netherlands, and the kinds of treatment they sought was undertaken in 1999. Qualitative data were collected from 12 women with primary or secondary infertility through narratives and 20 key informant interviews. The women described seeking treatment for infertility in Ghana, the Netherlands and other European countries, included use of infertility drugs, surgery, donor insemination and in vitro fertilisation. Illegal migrant women are not entitled to treatment paid by the national health system, and being of low income they cannot afford to pay directly for this or to obtain private health insurance. Herbalists and spiritual healers in both Amsterdam and Ghana were regularly consulted, especially for their willingness to address the social and spiritual aspects of infertility. To produce a pregnancy where male infertility was suspected, transfer of sexual rights to another man in the husband/partner's family, or a healer or priest, was a practical remedy that kept male infertility hidden. This study revealed difficulties experienced in clinical settings due to language barriers and cultural differences. Ghanaian women living in the Netherlands need much more information on the causes of infertility and their options. PMID- 11424242 TI - Mortality related to sexuality in Africa. PMID- 11424243 TI - Images and representation of non-motherhood. PMID- 11424244 TI - Strategies for prevention of mother-to-child transmission of HIV. AB - Over 90 per cent of paediatric HIV infections are maternally acquired, most of these in sub-Saharan Africa. Mortality trends underscore the humanitarian and ethical obligation for urgent global action to protect children from HIV. With the adoption of anti-retroviral therapy in pregnancy, mother-to-child transmission rates have declined to 4-6 per cent in the USA and other industrialised countries. In low-resource settings, where most of the children are continuously being exposed to HIV, the cost of anti-retroviral therapy is prohibitive. Very few developing countries apart from Botswana, Thailand and Brazil have national policies for integration of preventive anti-retroviral therapy in antenatal clinics. This paper reviews anti-retroviral and non-anti retroviral interventions for prevention of mother-to-child transmission of HIV. To support the health of mothers as well, it supports the implementation of a comprehensive package of care in pregnancy and post-partum, such as access to antenatal and delivery services; anti-retroviral preventive therapy; malaria treatment; family planning; multivitamin, iron and folate supplementation; counselling on feeding options; post-natal care for the child and post-partum care for the mother, and calls for a strategy for advocacy, programme communication and community mobilisation. PMID- 11424245 TI - What's up with nonoxynol-9? AB - Advantage-S, a spermicide containing nonoxynol-9 (N-9), has recently been shown in a randomised, controlled trial sponsored by UNAIDS not to offer women protection against sexual transmission of HIV. The women in the trial, all sex workers, reported having an average of 3.6 partners per day and about 70 coital acts per month during the study period. At the end of the trial, the Advantage-S group was found to have a higher incidence of new HIV infections than the control group, who were using a vaginal lubricant. Two other recent studies have found that low dose N-9 products do not provide protection against bacterial STDs such as gonorrhoea and chlamydia--a finding that contradicts earlier wisdom based on less rigorous studies. Contrary to certain media headlines, however, the results of these trials do not suggest that effective microbicides cannot be developed, only that Advantage-S, which was developed as a spermicide and not as a microbicide, is probably not one of them. The failure of Advantage-S to show protection in the UNAIDS-sponsored trial underscores the urgent need to accelerate research to develop a safe and effective topical microbicide. PMID- 11424246 TI - HIV/AIDS, sexual and reproductive health at AIDS 2000, Durban. AB - The 13th International AIDS Conference in Durban was a milestone in many ways, the most important being that it took place in southern Africa and brought together many more people from the African region with others from developing countries internationally than any previous conference. This should be repeated often and soon. For women, however, reproductive and sexual health issues were by and large not visible on the main conference agenda, except for prevention of mother-to-child transmission and sexually transmitted disease (STD) interventions, including STD prevalence, efforts at treatment and the effects of treatment on HIV transmission. An analysis of all the abstracts on the conference CD-Rom (including both presentations and posters, a total of almost 5,000 abstracts), using 25 reproductive and sexual health keywords, turned up much valuable work, and some new developments worth knowing about. These are summarised here, followed by condensed versions of some of the most interesting abstracts. PMID- 11424247 TI - Advocacy for sexual and reproductive health: the challenge in India. AB - Six years after the Indian government affirmed its commitment to the principles of the 1994 International Conference on Population and Development, there is still a limited understanding of the concept of reproductive and sexual health among policymakers, programme managers and the public in India. Despite some progressive changes, there is a continuing focus on stabilising population growth rates and lack of unity of goals among women's rights advocates, service providers and policymakers. Advocacy efforts need to begin focusing on turning progressive reproductive health policies into concrete programmes on the ground, and continue to push for progressive policies in uncharted areas such as domestic violence. Those who implement programmes need to work with potential allies such as women's groups, development groups, health workers' associations and the media, who all need to be brought on board. What continues to be missing are policies and programmes that promote not only health but also rights and the empowerment of women. Without a strong focus on the links between these, India's reproductive health policies and programmes may become like 'grass without roots'. PMID- 11424248 TI - National crisis, supranational opportunity: the Irish construction of abortion as a European service. AB - In the late 1980s, the anti-abortion movement successfully sought injunctions against pregnancy counselling centres and students' unions in Ireland, preventing them from distributing information on how to obtain an abortion abroad. One of the defensive arguments that the students' unions employed was to claim that the distribution of abortion information was protected as an aspect of the free movement of services under European Community law. This paper addresses the implications of categorising abortion as a supranational economic service for feminist legal strategy. The advantages of categorising abortion as a service to which women have access as consumers are that it legitimates abortion and it provides a new strategy for making abortion claims. The disadvantages are that a woman's legal interest in abortion is based on her capacity to buy the service, fetal life is rendered devoid of value, and the service supplier has as much say about the abortion transaction as the woman consumer. If feminist legal strategy is to successfully use the legal construction of abortion as an economic service, it must work to minimise such negative implications. PMID- 11424249 TI - Reproductive rights of Egyptian women: issues for debate. AB - Because the definitions of reproductive health and reproductive rights are so broad, it has been possible in Egypt and elsewhere for one or other issue on the reproductive health agenda to be prioritised to the detriment or exclusion of others, and a comprehensive approach avoided. Reproductive health and rights language was introduced in Egypt around the time of the International Conference on Population and Development in Cairo in 1994. From the moment these concepts were translated into Arabic, activists, scholars and women themselves discussed and debated their meanings and questioned whether Egyptian culture permitted a rights approach of this kind and whether reproductive rights were actually perceived as such by Egyptian women. This paper discusses the language of reproductive rights in relation to cultural specificity in the Egyptian context, Egyptian women's perceptions of reproductive rights, the role of tradition and religion, and the ways in which reproductive rights are taken up under Egyptian law. It analyses some of the arguments used against reproductive rights on these grounds. PMID- 11424250 TI - The value of advocacy in promoting social change: implementing the new Domestic Violence Act in South Africa. AB - South Africa's first democratic government passed the Domestic Violence Act (DVA) into law in 1998 as part of local and international commitments to protecting the human rights of women. Although the Act was welcomed as groundbreaking legislation, delays in implementing it led to increasing frustration. This paper describes an advocacy campaign conducted by the Soul City Institute for Health and Development Communication in partnership with the National Network on Violence against Women, to ensure the effective implementation of the DVA. Lessons from the campaign stress the importance of coalition building to draw on diverse strengths, and the use of a combination of advocacy tools, including lobbying, media advocacy and social mobilisation to achieve campaign goals. Given the critical role NGOs dealing with victims/survivors of domestic violence and the justice system played in lobbying for change and drafting the new law, their exclusion from the implementation process was ironic. While many advocacy efforts focus on the development of policy and legislation, ongoing efforts are needed to ensure effective implementation, the commitment of adequate resources and monitoring to identify gaps and propose new solutions. Our experience highlights the important role of policy advocates in connecting the multiple streams at play in the policy and legislative arena. PMID- 11424251 TI - Reproductive rights, advocacy and changing the law. PMID- 11424252 TI - Abortion in Brazil: legislation, reality and options. AB - Abortion is illegal in Brazil except when performed to save the woman's life or in cases of rape. This paper gives a brief history of parliamentary and extra parliamentary efforts to change abortion-related legislation in Brazil in the past 60 years, the contents of some of the 53 bills that have been tabled in that time, the non-governmental stakeholders involved and the debate itself in recent decades. The authorities in Brazil have never assumed full public responsibility for reproductive health care or family planning, let alone legal abortion; the ambivalence of the medical profession is an important obstacle. Most politicians avoid getting involved in the abortion debate, but the majority of bills in the 1990s have favoured less restrictive legislation. Incremental legislative and health service changes could help to improve the situation for women. Advocacy is probably the most important action, to promote an environment conducive to change. Clandestine abortion is a serious public health problem in Brazil, and the inadequacy of family planning services is one of the causes of this problem. The solutions should be made a priority for the Brazilian public health system. PMID- 11424253 TI - Making legal abortion available in Brazil: partnerships in practice. AB - This article describes the participation of feminist groups who work in the area of women's reproductive health and rights in campaigns for the provision of legal abortion in public hospitals in Brazil. Brazilian criminal law permits therapeutic abortion in cases where pregnancy is the result of rape or poses a serious risk to the life of the woman. Today, as a result of the combined efforts of feminists, health professionals and policymakers, more than 20 hospitals in Brazil are officially permitted to perform therapeutic abortions within the existing law. A model programme has also been developed to train service providers to do legal abortions, where the agreement of a hospital board can be obtained. This training has also improved care for illegally obtained, incomplete abortions in those hospitals but not in hospitals where doctors have not been trained. Problems with lack of access and concerns about the lack of public acceptance of abortion remain. Women not only need the right to abortion but also more services and health professionals who are trained to perform abortions across the whole country. PMID- 11424254 TI - The reality of unsafe abortion in a rural community in South India. AB - Though the law in India has permitted medical termination of pregnancy on broad legal grounds for over two decades, unsafe abortions carried out by unqualified providers show no signs of decreasing. A community-based study was undertaken in rural South India to determine the prevalence of induced abortion, women's reasons for seeking abortion, who was providing abortions and whether the procedures were safe or unsafe. A cross-sectional study design was used that included focus group discussions with 88 women and a quantitative survey with 195 married women who had a birth interval of two or more years since their last pregnancy. There was a high prevalence of induced abortion (28 per cent) among the study population, mainly among women who were not using contraception. Most abortions were carried out in the first trimester of pregnancy and unqualified practitioners performed 65 per cent of terminations. The preference for illegal, untrained providers in a country where abortion is legally available exposes the ambiguity in the status of abortion and how inadequacies in legal service provision have served to promote and sustain unsafe providers and practices. An integrated approach to family planning and reproductive health which includes abortion is imperative if the reproductive health status of Indian women is to be improved. The poor utilisation of existing government facilities suggests the need for improving the quality of services, expansion of abortion facilities and the introduction of safer methods of abortion. To do this, a re-examination and re-framing of aspects of the 1971 abortion law is needed. PMID- 11424255 TI - Quietly working for school-based sexuality education in Mexico: strategies for advocacy. AB - In the past decade, there have been dramatic changes in Mexico in public opinion and support for sexuality education. The growing recognition that young people are sexually active before marriage and need information in order to make responsible choices is the result of intensive health promotion efforts by public and private entities. This paper is about the work of a Mexican non-governmental organisation, IMIFAP, which has designed and evaluated sexuality education programmes for children from pre-school age through adolescence and for their parents, based on formative research. IMIFAP has also been advocating with others for the wider provision of sexuality and life skills education in schools, work that was rewarded in 1998 when the Ministry of Education announced its support for comprehensive sexuality education in the standard school curriculum. IMIFAP's strategies for formulating, implementing and evaluating an integrated programme of sexuality education in a developing country, and advocating for its inclusion in national curricula, could serve as a model which others can adapt. PMID- 11424256 TI - Unethical ethics? Reflections on intercultural research practices. AB - The notion that there is a universal ethics is commonly supposed, but less often explicitly discussed, in protocols for ethical procedures in research. In this article, the authors reflect on their action-research with women farmers in a Bolivian highland province. Their project aims to propose ways in which local health services could better serve these women's expressed sexual and reproductive health needs. A series of field experiences led the authors to question the adequacy of pre-established institutional protocols for informed consent. Cultural understandings in this context made for situations where supposedly ethical procedures led to unethical effects. While recognising the value of precautionary measures to avoid abuse of research subjects, the authors challenge the assumption of subjects' essential vulnerability in fieldwork relations. They found that they too, as researchers and outsiders to the community, could be assigned less than adult status by research subjects seeking empowerment in their own terms. The paper concludes that consent protocols, rather than relying on standardised procedures, should provide flexible alternatives to facilitate negotiation with subjects about whether and how they will participate at different stages of a research project. PMID- 11424257 TI - A consumer trip into the world of the DALY calculations: an Alice-in-Wonderland experience. AB - For a consumer advocate, entering the world of the disability-adjusted life years (DALYs) calculations was a surreal experience. What began with the noble aim of working out how to overcome the tyranny of death as the only way of 'measuring' health, has led to an exercise where people weigh the relative value of health conditions without death as a feature. However, life-threatening diseases, if they no longer carry the threat of death, are no longer themselves. These valuations of 'altered health states' then, can become absurd. While data often fail to measure real life very well, and distort as well as illuminate, the ranking of disease in this way is particularly problematic. It has been said to disadvantage the poor, the very young and very old, and women. It privileges short-term interventions over prevention and long-term strategies. Can this really improve decision-making about resource allocation in health? PMID- 11424258 TI - Is it feasible or desirable to measure burdens of disease as a single number? AB - This paper questions the feasibility and desirability of creating a composite health index, such as the DALY (disability adjusted life years), that aggregates information about both mortality and morbidity into one number, to represent the burden of disease. Despite the expressed intention not to include contextual factors in the description of health states, in practice, doing so is unavoidable when trying to construct a health indicator. Including contextual factors leads to severe problems, however, regarding validity and reliability, and these are increased when evaluations of the described health states are made. This renders little confidence in DALYs as a measurement of the burden of disease. Using cost effectiveness analyses and DALYs as tools in priority setting might have unwanted consequences, and this is not the only method of achieving economic efficiency. The use of this method might also serve to relieve politicians of the responsibility for resource allocation in the health sector. An alternative to a composite index is better and more comprehensive epidemiological data on sexual and reproductive health mortality, morbidity and diseases rather than the better use of health indicators of questionable validity and reliability. These kinds of data are now being collected in connection with the Global Burden of Disease Project. It should be up to the decisions-makers to allocate existing resources based on a balancing of the many good purposes and goals available. PMID- 11424259 TI - Human rights, reproductive health and economic justice: why they are indivisible. PMID- 11424260 TI - Microbicides 2000: report of an international conference, 13-16 March, Washington DC. AB - 'Microbicide' is the name for a number of new products currently under research which could become a new class of barrier method for use in the vagina and rectum for protection against HIV, prevention and/or treatment of other sexually transmitted diseases and/or act as a contraceptive. This is a summary report of this scientific conference, at which basic science, clinical trial efficacy and design, and ethical, behavioural and public health issues were on the agenda. There are many potential products but few have yet got beyond Phase I clinical trials. There is not likely to be an approved and available product for 5-10 years at best; finding a balance of low toxicity with high efficacy is the major challenge, given how rapidly HIV infection actually infects tissue. Phase III clinical trials of these products require new protocols and procedures. It was urged by many, and underscored by HIV positive women who were present that clinical trials must be treated as an opportunity to promote a total prevention package--STD treatment, voluntary HIV testing and counselling, condom use, practising safer sex--plus microbicides and contraception. Identifying effective products and making them available as soon as possible were obviously also a priority. How to do this--ethically and scientifically--were the main subject of the meeting. PMID- 11424261 TI - Men's attitudes towards a potential vaginal microbicide in Zimbabwe, Mexico and the USA. AB - Vaginal microbicides are being developed in an attempt to expand women's and men's options for protecting themselves against HIV and other sexually transmitted infections. Taking account of men's attitudes during product development and introduction could increase the likelihood that products are acceptable and used. To capture the perspectives of urban and rural men from a range of cultural settings, we conducted focus group discussions with taxi drivers and farmers in Zimbabwe, Mexico and the USA. These explored HIV/STD risk perception and prevention strategies, desirable characteristics of vaginal products and of a microbicide, and attitudes towards use of a potential product. Men were generally supportive of the idea of a microbicide; urban somewhat more than rural men. Most thought microbicides would be preferable to condoms though many raised concerns about potential side effects. The men wanted these products to be as inexpensive and readily available as condoms, and differed as to whether a woman should have permission from her partner to use it. For them to be widely used, the men thought these products must not only be safe and effective, but should also have no negative effect on sexual pleasure. When a product becomes available, introductory messages must explain the limits on its effectiveness and encourage use with condoms. Further research is needed on definitions of pleasurable sex and the ramifications of this for microbicide formulation, and on partner communication around issues of sexuality and prevention of infection. PMID- 11424262 TI - Menstrual dysfunction: a missed opportunity for improving reproductive health in developing countries. AB - The field of population has undergone a paradigm shift to a broader focus on reproductive health, which recognises women's self-perceived health needs. Investigations in various countries reveal that menstruation is a primary concern of women. Yet sparse attention has been paid to understanding or ameliorating women's menstrual complaints. We propose including the management of menstrual complaints as part of reproductive health programming. Next steps should include further quantitative and qualitative research to understand the prevalence, determinants and consequences of menstrual dysfunction; developing appropriate protocols and low-cost interventions for diagnosis and treatment of menstrual morbidity and training of health care workers in resource-scarce settings; and developing educational interventions to facilitate women's understanding of normal menstrual function and variability as well as of the types, causes and appropriate treatments for menstrual dysfunction. PMID- 11424263 TI - Disclosure of HIV status and human rights: the duties and responsibilities of couples, medical professionals, family members and the state. AB - This case history is about a young husband in India who became HIV positive and did not inform his wife. This man was working for a company in the city of Mumbai when he became ill with tuberculosis and was diagnosed with AIDS. This information was not disclosed to his 21-year-old wife, who was living with her parents-in-law in a rural area of South Kerala. He came home to recover and, supported by his parents' wishes, wanted his wife to become pregnant, even though she had asked that they wait until he had recovered. She came to know that her husband had AIDS and the dangers it posed only when she was eight months pregnant and he was seriously ill. Her husband died some three months after the baby was born and she went back to her parents' house, where she and the baby were living at the mercy of others. The contributors to this multi-authored Roundtable discuss the various human rights issues and the duties and responsibilities involved--not only of spouses and partners to each other but also of medical professionals, family members and the state--which arise from such a case. PMID- 11424264 TI - Promoting reproductive and sexual health in the era of sector-wide approaches (SWAPs). PMID- 11424265 TI - Transforming family planning programmes: towards a framework for advancing the reproductive rights agenda. AB - Recent international agreements call for the transformation of family planning programmes from a focus on demographic goals to the promotion of health and rights objectives. But the practical implications of this agenda for current and future programmes remain unclear. Public health resources are devoted to preventing illness and reducing the prevalence and incidence of disease across a population. Human rights methodologies focus on protecting the rights of individuals, and on the right to health and health care. Both of these approaches need to be re-thought and reconciled on a practical level to promote rights-based health programmes. Applying a rights framework to reproductive health programmes means, among other things, focusing as much on the process as on the outcome, incorporating efforts to address the gender and power dimensions of reproductive and sexual decision-making into every level of programme, and focusing on building a sense of entitlement among both the seekers and the providers of services. It also means moving beyond a focus only on the technical quality of clinic-based services to incorporate the ethos of a rights perspective at every level. Political, institutional, and technical barriers to the realisation of the reproductive health and rights agenda include national level politics, lack of capacity within civil society, and lack of transparency of institutional actors. PMID- 11424266 TI - Living with infertility: experiences among Urban slum populations in Bangladesh. AB - This paper explores the perceived causes of infertility, treatment-seeking for infertility and the consequences of childlessness, particularly for women, among a predominantly Muslim population in urban slums of Dhaka in Bangladesh. In-depth interviews were conducted with 60 women and 60 men randomly selected from Urban Surveillance System clusters of the International Centre for Diarrhoeal Disease Research, Bangladesh. Case studies of 20 self-perceived infertile women who had previously participated in a study on the prevalence of sexually transmitted diseases and other reproductive tract infections were taken, and three traditional healers were interviewed as key informants. In both groups of respondents, the leading causes of infertility were perceived to be evil spirits and physiological defects in women and psychosexual problems and physiological defects in men. Herbalists and traditional healers were considered the leading treatment option for women, while for men it was remarriage, followed by herbalists and traditional healers. Childlessness was found to result in perceived role failure, with social and emotional consequences for both men and women, and often resulted in social stigmatisation of the couple, particularly of the woman. Infertility places women at risk of social and familial displacement, and women clearly bear the greatest burden of infertility. Successful programmes for dealing with infertility in Bangladesh need to include both appropriate and effective sources of treatment at community level and community-based interventions to demystify the causes of infertility, so that people know why infertility occurs in both men and women and and where best to seek care. PMID- 11424267 TI - Easing the pain: pain management in the treatment of incomplete abortion. AB - Although there have been significant improvements in post-abortion care programmes around the world, improving pain management has remained a significant challenge. The introduction of manual vacuum aspiration (MVA) has led to many positive changes in programmes, but the guidelines for pain control have generally been vague. Women are often treated with no pain control or in some cases receive too much pain medication. There are many factors contributing to this situation, including: the belief that women who have induced an abortion should be punished, the idea that pain control is unnecessary, the lack of availability of drugs and inadequate training and/or skills of providers. This paper argues for a greater focus on this important element of quality of care and for clearer guidelines on pain management during treatment of incomplete abortion with MVA. This includes the provision of analgesics immediately before the procedure, counselling and reassurance during the procedure and local anaesthesia when necessary. PMID- 11424268 TI - Adolescent girls with illegally induced abortion in Dar es Salaam: the discrepancy between sexual behaviour and lack of access to contraception. AB - This article reports on a study of induced abortion among adolescent girls in Dar es Salaam, Tanzania, who were admitted to a district hospital in Dar es Salaam because of an illegally induced abortion in 1997. In the quantitative part of the study, 197 teenage girls (aged 14-19) were asked for socio-economic details, contraceptive knowledge/use, age at first intercourse and number of sexual partners. In the qualitative part, 51 teenage girls were interviewed in-depth about their relationships with their partners, sexual behaviour, contraceptive use and reasons for non-use, and why they became pregnant. The girls were sexually active at an early age and having sex mainly with men older than themselves. Although most of the girls were in love with and enjoyed sex with their partners, they also entered these relationships to obtain money or gifts in exchange for sex. Most were not using contraception or condoms though they were also at risk of STDs and HIV. These girls were getting pregnant expecting their boyfriends to marry them, or because they did not think they could become pregnant or failed to use contraception correctly. Most adolescent girls are not aware of the 1994 Tanzanian policy that gave them the right to seek family planning services and in practice these services are not being provided. There is a need for youth-friendly family planning services and to make abortion safe and legal, in order to reduce unwanted pregnancies and abortion-related complications and deaths among adolescent girls. PMID- 11424269 TI - AIDS into the 21st century: some critical considerations. AB - The story of AIDS in the 21st century is likely to be dominated by heterosexuals in Africa and injecting drug users around the world. HIV infection has persisted and grown because people do not like to recognise, much less talk about, the behaviours that spread the virus. Drug injection remains the leading cause of HIV infection in in the countries of the former Soviet Union, the northeastern states of India, the USA, Western Europe, China and parts of the Middle East. In Africa, where the overwhelming majority of infections occur during unprotected sex, a high proportion of girls are infected with HIV during their teens and before marriage. Industrialised countries responded with massive prevention campaigns, open discussion of the potential dangers of unprotected sex, and aggressive condom promotion. In the few developing countries that have taken similar action to contain the epidemic--Thailand, Uganda and Senegal--the initial leadership came from senior politicians. As HIV prevalence rises in a population, the chance of someone encountering an infected partner close to the beginning of their sexual life also rises. It is therefore crucial to reach people with appropriate preventative interventions before they first have sex. However, it is not realistic to expect political commitment to as sensitive a problem as AIDS from any government that has less than a wholehearted commitment to the basic health and welfare of its people. The first challenge in the fight against AIDS, as in the fight for development in general, is to support good government. PMID- 11424270 TI - Safe sex, women's reproductive rights and the need for a feminist movement in the 21st century. PMID- 11424271 TI - The impact of HIV on family size preference in Zambia. AB - This paper examines the way in which concerns about HIV infection are affecting thinking in Zambia about preferred number of children. It draws on research on the impact of HIV/AIDS in peri-urban and rural households in 1995, based mainly on in-depth interviews with 65 of 300 people who were initially surveyed. In spite of high levels of anxiety about AIDS in these communities, risk from HIV was not always associated with the act of conceiving children, nor did this association necessarily influence actual behaviour or family size preferences. In some cases, however, the threat of contracting HIV had led to a decision to have fewer children. Many also worried about leaving orphans for others to look after and the costs which might be incurred in taking over the care of orphans left by others. A related reason for limiting fertility was the hope that orphaned children would be better cared for if there were fewer of them. Greater access to contraceptives, and specifically to condoms, is an important element in supporting women's efforts to protect themselves, and men also need to be involved in strategies for mutual protection. In both communities, however, there was a shared sense of limited control, not just over fertility, but also over the wider economic and health environment. An understanding of the complexity of these factors is essential for intervention programmes intended to enhance women's reproductive rights and support their fertility choices so as to ensure greater protection against HIV/AIDS. PMID- 11424272 TI - Preventing mother-to-child transmission of HIV in developing countries: recent developments and ethical implications. AB - Various regimens of antiretroviral (ARV) therapy during pregnancy and labour have been found to be effective in reducing the risk of mother-to-child transmission of HIV. Cost and late identification of women with HIV infection during pregnancy in many developing countries have been the impetus to study inexpensive, short course ARV regimens. Recently, it was shown that a single dose of nevirapine given orally once during labour to the mother and once to the infant greatly reduces the risk of HIV transmission. As a result, it has been proposed that in high HIV prevalence areas, this drug regimen be offered routinely to all pregnant women and their infants, without the need for an HIV test. This is seen as a cost effective alternative to trying to make voluntary HIV testing and counselling universally available to pregnant women, which would require improved antenatal uptake and care, high uptake of HIV testing and high rates of return to learn results before women could make decisions regarding ARV prophylaxis. The ethical dilemmas arising from both these options are currently under debate, against a backdrop of concerns about breastfeeding and breastmilk substitutes, what to do about the increasing numbers of AIDS orphans and how to prevent HIV transmission to women in the first place. PMID- 11424273 TI - Ethical and methodological issues in conducting research on gender-based violence in Southern Africa. AB - International experience of carrying out research on violence against women among women who may have experienced such violence and among men who may have been violent has raised a number of ethical issues, which appear in sharp focus. This paper is based on the collective experiences derived from involvement in three large gender-based violence surveys in South Africa and one in Zimbabwe. These are broadly concerned with the safety of researchers and research subjects from violence arising because of the research project, risks of traumatisation of both respondents and researchers as violent experiences are recounted; impact of work on violence as an issue in researchers' own relationships; risks of under reporting the extent of violence experienced or perpetrated; and the need for research efforts to be followed up in ways beneficial to women experiencing violence, such as increased support, public information, and law and policy changes. This paper discusses how these issues have pertained and been addressed in research on gender-based violence in Southern Africa and consider lessons for this and other sensitive areas of reproductive and sexual health research. PMID- 11424274 TI - By the way, doctor. NOw that pollen season is underway, I need a decongestant. The one I've used for years was taken off the market because it contained PPA. What is PPA, and why is it a problem? What can I use instead? PMID- 11424275 TI - By the way, doctor...I'm a few years past menopause, and I take a statin because of high cholesterol. My doctor has broached the subject of whether I might need to take something else to lower my risk for osteoporosis, but I've read that statins help prevent that problem. Can't I tell the doctor I don't need another pill? PMID- 11424276 TI - By the way, doctor...I used a diet pill--fenfluramine--a few years ago, but stopped as soon as I heard about it causing heart-valve problems. My doctor has been doing echocardiograms on me every six months. So far, everything looks good. How long do you think I need to keep on getting these checkups? PMID- 11424277 TI - Does myalgic encephalomyelitis exist? PMID- 11424278 TI - ABC of secondary prevention. PMID- 11424279 TI - Index of suspicion. Case 2. Diagnosis: intussusception. PMID- 11424280 TI - Index of suspicion. Case 3. Diagnosis: Hypercalcemia. PMID- 11424281 TI - Misoprostol and the politics of fear. PMID- 11424282 TI - Research in less-developed countries. PMID- 11424283 TI - New use for a prep sponge. PMID- 11424284 TI - Lab portion of OSHA Exposure Control Plan for Bloodborne Pathogens. PMID- 11424285 TI - The OSHA lowdown on occupational exposure to bloodborne pathogens. PMID- 11424286 TI - Implementing a standardized POCT coagulation system. PMID- 11424287 TI - Multitude of professional organizations offers choices and benefits for laboratorians. PMID- 11424288 TI - Exclusive privileges: use as a competitive tool. PMID- 11424289 TI - Internal Revenue Service issues additional guidance regarding exempt hospitals' compliance with "operational test". PMID- 11424290 TI - Illinois appellate courts disagree on the validity of physician non-compete agreements. PMID- 11424291 TI - Considerations in the spin-off/dissolution of a medical group. PMID- 11424292 TI - Trust, the strategic imperative: how to successfully build physician-system partnerships. PMID- 11424293 TI - EEG frequency and phase coupling during human information processing. AB - Neuronal activity during information processing is represented by oscillations within local or widespread neuronal networks. These oscillations may be recorded by the EEG (electroencephalogram). The oscillatory interaction between neuronal ensembles may be at one single frequency or at different frequencies due to non linear coupling. The investigation of momentary coherence and phase enables the examination of synchronized oscillatory network activity during fast-changing cognitive processes. On this basis information transfer from occipital areas towards frontal areas could be described during processing of visual presented words. Non-linear phase coupling between oscillations with different frequencies during memory processing was detected by means of cross-bicoherence. PMID- 11424294 TI - Analytical and graphical methods to model the association between family history and disease from a case-control study. AB - Subjects within a family have a high probability of sharing environmental factors as well as genes. Without any previous hypothesis, familial aggregation cannot be attributed to genetic or environmental factors. In epidemiology a common measure of familial aggregation is the odds ratio (OR) evaluated from case-control studies which gives a measure of the increased risk associated with particular aspects of the family's history of disease. An analytical method is presented to model the OR of case-control studies in the presence of risk factors, genetic or environmental, that are correlated between relatives. A graphical approach that can be used to link the parameters of the model with the values of the OR is also described. The method is illustrated using the example of familial aggregation in colorectal cancer. PMID- 11424295 TI - Flexible two-stage designs: an overview. AB - In this overview we introduce the basic ideas behind a new flexible approach in sequential designs. The different concepts based on two-stage combination tests and conditional error functions are brought together. We sketch the construction of p-values, confidence intervals, and median unbiased estimates. Finally, recursive combination tests are introduced which extend the flexibility to the choice of the number of interim analyses. PMID- 11424296 TI - A model and application for estimating completeness of registration. AB - Completeness of population-based registration systems is recognized to be an important aspect of the quality of information in registries which has to be examined. In this paper a model is presented which was used to estimate completeness of reporting of Down syndrome data notified to the Styrian Malformation registry between 1985-92. The model introduced is based on the two source capture-recapture method allowing for time-varying parameters. For estimation of the parameters a discrete-time filtering algorithm was developed. For the used data set, an estimate of completeness derived from this model was in good agreement with an independent estimator based on demographic data and maternal age-specific Down syndrome risks whereas the usual two-source capture recapture method gave a higher estimate. PMID- 11424297 TI - Childhood leukaemia clustering--fact or artefact? AB - There is a general belief that the clustering of childhood leukaemia is a widespread phenomenon and that it provides evidence for appreciable environmental influence on the incidence of the disease. We discuss this issue critically, identifying different kinds of clustering and their possible aetiological mechanisms and examining some analyses of British data. We argue that, in some cases, analyses have been used which lead to dubious conclusions, and that, allowing for multiple testing and anecdotal reporting, the total evidence for clustering is at best weak. PMID- 11424298 TI - Age-related reference regions for longitudinal measurements of growth characteristics. AB - Most studies on age-related reference centiles published up to now have adopted a strictly cross-sectional perspective. Clearly, the results of studies of that type do not provide a tool for the diagnostic assessment of whole series of measurements taken sequentially over time in the same individual. In this paper, the approach of Wellek & Merz (1995) to the construction of age-dependent reference ranges for cross-sectional measurements is generalized in such a way that data sets containing time series of arbitrary length varying between subjects can be accommodated. Since repeated measurements on the same subject are typically correlated, the regression function to be used as the central line for the reference band eventually obtained is determined by fitting a nonlinear mixed model describing the dependence of conditional means on age by growth functions of the same class we proposed in the case of cross-sectional data. Estimation of the parameters of this mixed model is done in a way closely related to the population-averaged GEE approach by Zeger et al. (1988). Given the regression line, the reference band is constructed by means of an iterative procedure guaranteeing that the proportion of observed profiles which nowhere leave the band, has some prespecified value (frequently set equal to 90% in practice). The approach is illustrated with two examples taken from child psychiatry and prenatal sonography. PMID- 11424299 TI - Design issues and sample size when exposure measurement is inaccurate. AB - Measurement error often leads to biased estimates and incorrect tests in epidemiological studies. These problems can be corrected by design modifications which allow for refined statistical models, or in some situations by adjusted sample sizes to compensate a power reduction. The design options are mainly an additional replication or internal validation study. Sample size calculations for these designs are more complex, since usually there is no unique design solution to obtain a prespecified power. Thus, additionally to a power requirement, an optimal design should also fulfill the criteria of minimizing overall costs. In this review corresponding strategies and formulae are described and appraised. PMID- 11424300 TI - Survival analysis with time-varying relative risks: a tree-based approach. AB - A tree-based method for estimating time-varying effects of baseline patient characteristics on survival is introduced. A Cox-type model for censored survival data is used in which the time-varying relative risks are modelled as piecewise constants. The tree method consists of three steps: 1. Growing the tree, in which a fast algorithm using maximized score statistics is utilized to determine the optimal change points; 2. A pruning algorithm is applied to obtain more parsimonious models; 3. Selection of a final tree, which may be either via bootstrap resampling or based on a measure of explained variation. The piecewise constant model is more suitable for clinical interpretation of the regression parameters than the more continuously time-varying models (spline, loess, etc.) that have been proposed previously. PMID- 11424301 TI - Deficits and remedy of the standard random effects methods in meta-analysis. AB - The random effects model is often used in meta-analyses. A corresponding significance test based on a normal approximation has been established. Its type I error is derived in this article by theoretical considerations and computer simulations. The test can be conservative as well as unacceptably anti conservative. The anti-conservatism increases with the increasing number of patients and the decreasing number of studies. A modification is proposed, which keeps the nominal level asymptotically as the number of patients approaches infinity. Simulations show that the modified test is often conservative, but its conservatism is small in those situations where the standard test is highly anti conservative. PMID- 11424302 TI - Information processing in healthcare at the start of the third Millennium: potential and limitations. AB - The 21st century is said to be a century of the information society. We should be aware that continuing progress in information processing methodology (IPM) and information and communication technology (ICT) is changing our societies, including medicine and health care. At the start of the third Millennium we should ask ourselves, what progress can we expect from modern IPM/ICT for healthcare in the coming decade, what concerns does the information society have to face, and what steps have to be taken. These questions were addressed by clinicians, researchers and industrial representatives in a panel discussion at the joint conference ISCB-GMDS-99 of the International Society of Clinical Biostatistics and the German Society for Medical Informatics, Biometry and Epidemiology. Important aspects raised by the panelists and in the subsequent discussion were: (1) the main goal of expanding IPM/ICT should be to further improve quality of care, while maintaining reasonable costs; (2) with the support of modern IPM and ICT the boundaries between inpatient and outpatient care will fade away enabling a more efficient, patient-centered health care; (3) cooperation between health-care professionals will increase; there will be different ways of communication between them and with the patient, including modern ICT and the Internet; (4) society must be concerned with achieving equal opportunities in being informed about and in using new ICT; (5) misuse of data will remain a serious problem and can become an obstacle to progress. PMID- 11424303 TI - Digital Libraries and Recent Medical Informatics Research. Findings from the IMIA Yearbook of Medical Informatics 2001. AB - The Yearbook of Medical Informatics is published annually by the International Medical Informatics Association (IMIA) and contains a selection of recent excellent papers on medical informatics research (http://www.med.uni heidelberg.de/mi/yearbook/index.htm). The special topic of the just published Yearbook 2001 is "Digital Libraries and Medicine". Digital libraries have changed dramatically and will continue to change the way we work with medical knowledge. The selected papers present recent research and new results on digital libraries. As usual, the Yearbook 2001 also contains a variety of papers on other subjects relevant to medical informatics, such as Electronic Patient Records, Health Information Systems, Health and Clinical Management, Decision Support Systems, Education, as well as Image and Signal Processing. This paper will briefly introduce the contributions covering digital libraries and will show how medical informatics research contributes to this important topic. PMID- 11424304 TI - ISCB-GMDS-99. PMID- 11424305 TI - A randomized evaluation of a computer-based nursing documentation system. AB - A two-month randomized, controlled trial based on 60 patients has been performed on a ward of the Department of Psychiatry at Heidelberg University Medical Center, Germany, to investigate the influence of computer-based nursing documentation on time investment for documentation, quality of documentation and user acceptance. Time measurements, questionnaires, documentation analysis and interviews were used to compare patients documented with the computer-based system (PIK group) with the control group (patients documented with the paper based system). The results showed the advantages and disadvantages of computer based nursing documentation. Time needed for nursing care planning was lower in the PIK group. Some formal aspects of quality were considerably better in the PIK group. On the other hand, time required for documentation of tasks and for report writing was greater in the PIK group. User acceptance increased significantly during the study. The interviews indicated a positive influence of PIK on the cooperation between nurses and physicians. PMID- 11424306 TI - The need for open geographical information systems in medicine. AB - This paper presents a promising approach of using component-oriented geographical information systems (GIS) to facilitate improved medical and ecological applications. The integration of administrative geographical and medical data (map overlay) allows sophisticated and advanced analysis. This can serve as a basis to detect undiscovered relationships. We hope to increase the usage of geographical information by the creation of a general framework. This approach is mainly supported by two advances in recent years. Geographical data (e.g. administrative areas) is up-to-date, consistent and covers Germany completely. In addition, enormous technological improvements in the area of high-bandwidth networks and component-based software development have been made. These advances allowed to define our general framework for the integration of GIS functions into existing and already accepted applications. PMID- 11424307 TI - Simulation of hip operations and design of custom-made endoprostheses using virtual reality techniques. AB - In this paper a system for the virtual planning of hip operations with endoprosthetic reconstruction and its application in bone tumor surgery is described. The system enables the simulation of the operation and the construction of a custom-made implant depending on the chosen resection planes and the patient's anatomy. During the planning process integrated virtual reality techniques facilitate the interaction with the three-dimensional (3D) medical objects. Stereo viewing improves the perception of the 3D nature of bone structures and tumors. In comparison to conventional planning procedures, different operation strategies and their influence on the geometry of the custom made endoprosthesis can be easily compared. Furthermore, the combination of multi modal image information (CT and MR) enables an accurate 3D visualization of the bone tumor within the bone. PMID- 11424308 TI - Medical informatics and problem-based learning. AB - Problem-based learning (PBL) is an educational method that can be considered as an alternative to the traditional, discipline-based, approach to teaching. In this paper the characteristics of both approaches are discussed and compared. Some background knowledge concerning the principles that determine the success of instructional methods is presented. Finally, it is discussed how medical informatics can be taught in a PBL environment. PMID- 11424309 TI - A realistic model of human structure from the visible human data. AB - The computer-based 3D models of the human body reported to date suffer from poor spatial resolution. The Visible Human project has delivered high resolution cross sectional images that are suited for generation of high-quality models. Yet none of the 3D models described to date reflect the quality of the original images. We present a method of segmentation and visualization which provides a new quality of realism and detail. Using the example of a 3D model of the inner organs, we demonstrate that such models, especially when combined with a knowledge base, open new possibilities for scientific, educational, and clinical work. PMID- 11424310 TI - On the necessity of systematically planning clinical tumor documentation. AB - Tumor documentation is an important task for both clinical research and patient care. Documentation systems for these purposes have to be planned systematically and should be goal oriented. We applied the method of a so-called 'standardized documentation protocol' for systematically planning two documentation systems in oncology: one for the tumor center Heidelberg/Mannheim and the other for a nationwide project in the field of documentation and therapy planning in pediatric oncology. The method proved to be helpful in both cases even though the resulting documentation protocols are completely different and although they served different objectives. Therefore, the aim of the paper is to motivate and help medical informatics professionals to systematically plan other documentation systems using this method. PMID- 11424311 TI - Intranet meets hospital information systems: the solution to the integration problem? AB - Based on the results of an extensive market analysis and technology assessment, this paper summarizes the state of the art in web technology and compares it with the needs of integration in hospital information systems. Web technology and intranet have evolved to a core technology for distributed computing and distributed information systems. The advantage of platform independent distribution of information has led to euphoric visions of future integrated systems. However, true integration takes more than transparent access to heterogeneous information sources. To overcome the essential problems related to integration of autonomous systems it is important to find a common ontological basis for future component based systems. PMID- 11424312 TI - Peptic ulcer disease is strongly associated with Helicobacter pylori in east, west, central and South Africa. PMID- 11424313 TI - Dysphagia and oesophageal clearance after laparoscopic versus open Nissen fundoplication. A randomized, prospective trial. AB - BACKGROUND: An increase in postoperative dysphagia has been reported after laparoscopic fundoplication. Our aim was to compare laparoscopic Nissen-Rossetti fundoplication to open fundoplication regarding oesophageal clearance and dysphagia in a prospective, randomized study. METHODS: Twenty-eight consecutive patients with objectively observed gastro-oesophageal reflux disease referred to operative treatment were randomized to laparoscopic (13) or open (15) fundoplication. A standard formula was used in pre- and postoperative interview. Oesophageal clearance was measured by liquid bolus radionuclide transit before and 3 days, 1 month and 1 year after fundoplication. Endoscopy was done preoperatively and 1 year after the operation. RESULTS: Heartburn, regurgitation and ooesophagitis were cured with equal effectiveness (p = 0.001). New-onset dysphagia was observed in nine (69%) of the patients in the laparoscopic group and in nine (60%) in the open group during the first postoperative month. Food impaction occurred in four (31%) cases after laparoscopic and in two (13%) after open surgery (ns). One year after the operation, one patient (8%) in both groups had more than mild symptoms. Oesophageal radionuclide transit remained normal after open fundoplication, but after the laparoscopic procedure oesophageal clearance was disturbed--only one patient did not have a pathologic result during the first postoperative month. One year after the operation, clearance was normal. CONCLUSIONS: After laparoscopic operation, a tendency to more severe new onset dysphagia was observed, and oesophageal clearance was transiently disturbed. Efforts should be made to minimize postoperative swallowing and clearance disturbances after laparoscopic fundoplication in order to get the full value out of otherwise more rapid recovery. PMID- 11424314 TI - Evaluation of a rapid whole blood test to detect Helicobacter pylori infection in children. AB - BACKGROUND: Rapid, patient near tests for detecting antibodies against Helicobacter pylori are offered for clinical use. We evaluated the BM-Test (Boehringer Mannheim, Germany; identical to the Helisal Rapid Blood Test) in children with recurrent abdominal pain. METHODS: The BM-Test and 13C-urea breath test (UBT) were performed in 195 children (4-18 years). Symptoms were assessed using a 4-week diary. Upper endoscopy was performed in all UBT-positive children and in UBT-negative children with symptoms suggestive of organic disease. H. pylori status was considered positive if at least two of three methods (UBT, histology or rapid urease test) or culture were positive. RESULTS: After exclusion of children with previous H. pylori therapy (n = 8) and undetermined H. pylori status (n = 1), 61/186 (33%) children were H. pylori positive. The BM-Test in relation to H. pylori status revealed a sensitivity of 54%, specificity of 90%, a likelihood ratio of 5.2 for a positive, and of 0.4 for a negative test result. Accuracy of the test was independent of ethnicity, gender, age, family history for ulcer disease, frequency or severity of abdominal symptoms, epigastric tenderness, type of blood sampling (capillary versus venous) and DOB values of the UBT. In eight previously treated children, the test gave one false positive and three false-negative results. CONCLUSIONS: Almost half of H. pylori infected children and 10% of non-infected children were misclassified by the BM Test. False-negative results are not related to young age or certain ethnic groups. The poor performance makes the test unsuitable for epidemiological and clinical use in children. PMID- 11424315 TI - Effects of cyclooxygenase-2 inhibitor on gastric acid secretion in Helicobacter pylori-infected C57BL/6 mice. AB - BACKGROUND: Helicobacter pylori-associated body gastritis inhibits gastric acid secretion. The aim of this study was to determine the effects of H. pylori infection on gastric acid secretion and further determine whether cyclooxygenase 2 was involved. METHODS: C57BL/6 mice (n = 40) were inoculated with the Sydney strain of H. pylori. Control mice (n = 40) were treated with vehicle only. Half of the infected and control mice were fed an experimental diet containing etodolac (10 mg/kg/day) from 1 week after inoculation until the end of the experiment. Before, 12 and 24 weeks after inoculation, the gastric acid secretion, prostaglandin E2 (PGE2) levels in the gastric mucosa, and gastritis scores according to the updated Sydney system were determined. Immunohistochemical staining of COX-2 protein was also performed. RESULTS: No significant changes in gastric acid secretion, gastritis scores or PGE2 levels in the gastric mucosa were observed in uninfected groups with or without etodolac treatment during the study period. In the H. pylori-infected group without etodolac treatment, gastric acid secretion was significantly decreased with increases in PGE2 levels in the gastric mucosa 24 weeks after inoculation compared with the controls. Gastritis score for activity was significantly higher, and strong staining for COX-2 protein was observed in the H. pylori infected group. In the H. pylori-infected group with etodolac treatment, PGE2 in the gastric mucosa was decreased and acid secretion was restored to the same level as in the control group. CONCLUSION: One of the mechanisms by which H. pylori infection inhibits gastric acid secretion is increased release of PGE2 produced by COX-2, which is induced by H. pylori infection. PMID- 11424316 TI - Clarithromycin-based triple therapy for non-resistant Helicobacter pylori infection. How long should it be given? AB - BACKGROUND: We have previously reported mixed infection with wild-type (sensitive) and mutant (resistant) Helicobacter pylori strains using a PCR-based preferential homo-duplex formation assay (PCR-PHFA) to detect gene mutations associated with clarithromycin resistance. Half of the cases with mixed infection were determined as sensitive by conventional MIC assessment and yet failed to respond to clarithromycin-based therapy. The aim of this study is to assess the efficacy of clarithromycin-based triple therapy in patients infected exclusively with wild-type strains as determined by PCR-PHFA. METHODS: Ninety patients who had pure wild-type H. pylori infection were randomly assigned to receive clarithromycin (200 mg b.i.d.), amoxicillin (500 mg q.i.d.) and lansoprazole (30 mg b.i.d.) for either 5 days or 7 days (n = 48 and n = 42, respectively). The outcome of eradication was assessed by [13C] urea breath test. RESULTS: Eradication was achieved in 36/48 (75%) versus 39/42 (93%) by intention-to-treat analysis (P = 0.02), and in 36/45 (80%) versus 39/40 (98%) by per protocol analysis (P = 0.01), for the 5-day and 7-day protocols, respectively. Compliance and the incidence of untoward effects were similar in both groups. CONCLUSIONS: Seven-day administration is necessary and sufficient for the triple therapy with clarithromycin, amoxicillin and lansoprazole in patients with pure wild-type H. pylori infection. PMID- 11424317 TI - Inhibition of apoptosis in human neutrophils by Helicobacter pylori water-soluble surface proteins. AB - BACKGROUND: Helicobacter pylori infection in humans causes persistent neutrophil infiltration into the gastric mucosa. It is believed that a prolongation of neutrophil life-span could contribute to the pathogenesis of H. pylori infection. We therefore examined whether the water-soluble surface proteins of H. pylori can influence the apoptosis of neutrophils. METHODS: After neutrophils were incubated with H. pylori water extract (HPWE), neutrophil apoptosis was evaluated by TUNEL assay, Hoechst 33342 staining, electron microscopy and ELISA for cytosolic oligonucleosome-bound DNA for up to 48 h. To investigate the regulatory mechanisms of neutrophil apoptosis associated with HPWE, mRNA expression and protein production of Fas, Fas ligand (FasL) and tumor necrosis factor receptor 1 (TNF-R1) were analyzed by RT-PCR, ribonuclease protection assay, Northern blot and Western blotting. Cell surface expression of these death factors was also measured by flow cytometry. RESULTS: HPWE inhibited neutrophil apoptosis and cytotoxicity for up to 48 h. The mRNA and protein expression of FasL and the cell surface expression of Fas, FasL and TNF-R1 in HPWE-treated neutrophils were suppressed compared with the controls. CONCLUSION: The water-soluble surface proteins of H. pylori could suppress neutrophil apoptosis. This may be caused by the suppression of FasL expression in neutrophils and Fas, FasL and TNF-R1 expression on the surface of neutrophils. PMID- 11424318 TI - Long-term course in collagenous colitis and the impact of bile acid malabsorption and bile acid sequestrants on histopathology and clinical features. AB - BACKGROUND: Bile acid malabsorption is common in collagenous colitis, although long-term follow-up data on the impact of bile acids are limited. The aim was to study whether bile acid malabsorption is a permanent finding, with an impact on histopathology and clinical features in collagenous colitis. METHODS: The objective was to reinvestigate 27 patients with collagenous colitis > or = 3 years after index investigation. The clinical course was evaluated by means of an interview, a review of the hospital records and registration of symptoms over a period of 7 days. The patients were invited to undergo a repeat colonoscopy and 75SeHCAT measurement. Initial and follow-up data and 75SeHCAT values from 29 controls were compared. RESULTS: The median follow-up time was 4.2 (range 3-5.3) years. Twenty-two patients underwent a repeat 75SeHCAT test, 23 patients a colonoscopy and in 25 patients the clinical course could be evaluated. The 75SeHCAT values were abnormal in 32% at follow-up versus 44% at index, and the median retention value was 19% (range 2-69) versus 12% (range 0.5-41) (P = 0.024) although lower than in the control groups figure of 38% (range 8-91) (P < 0.005). Histopathology had improved independently of bile acid malabsorption, gender, smoking and autoimmune disease at follow-up. Four were normalized. Patients on bile acid binders had no significant change of histopathology. Four patients had recovered, seven displayed an intermittent course and 14 had continuous diarrhoea. CONCLUSIONS: Collagenous colitis and bile acid malabsorption seem to be associated yet independent disorders. The histopathology improves during the long-term course although only a few patients resolve. PMID- 11424319 TI - Oral microemulsion cyclosporine in the treatment of steroid-refractory attacks of ulcerative and indeterminate colitis. AB - BACKGROUND: To assess the efficacy of oral microemulsion cyclosporine (CyA Neoral) in the treatment of steroid-refractory attacks of ulcerative an indeterminate colitis. METHODS: In this non-randomized study on the use of oral microemulsion cyclosporine in steroid refractory ulcerative colitis, we used CyA Neoral in 10 patients suffering from ulcerative colitis and in 1 patient suffering from indeterminate colitis with a steroid-refractory attack. The initial dose was 7-7.5 mg kg-1 day-1 adjusted to maintain whole blood cyclosporine levels between 250 and 350 ng/mL, as measured by RIA using monoclonal antibodies. RESULTS: Nine patients presented a favourable response in a mean time of 3.6 days, that is, 81.8% of cases. One initial responder developed megacolon on the 11th day and another did not respond; surgical treatment was performed in both cases. The remaining nine patients, followed for a mean period of 14.6 months (2-36 months). Nine patients presented side effects, the most frequent being slight hand tremor with hypomagnesaemia, followed by hypertension, slight increase in creatinine and hirsutism. No one needed to withdraw from treatment, but the dose was lowered in three cases. CONCLUSIONS: Oral microemulsion cyclosporine is an effective drug in the initial management of patients suffering from a steroid-refractory attack of ulcerative and indeterminate colitis. Additional controlled studies with the new oral formulation are required to confirm these results. PMID- 11424320 TI - Staphylococcus aureus in inflammatory bowel disease. AB - BACKGROUND: The potential role of superantigens in inflammatory bowel disease (IBD), particularly Crohn disease, has been broached in studies of the functions of T cell receptors. Staphylococcal cells have been found in intestinal lymph follicles of IBDs. To clarify a role of staphylococcal superantigens in IBD, we attempted to determine whether Staphylococcus aureus could be detected in intestinal mucosa, including surgical specimens and lymph follicles of initial cases. METHODS: One-hundred-and-six colonic and ileal specimens were obtained from 38 Crohn disease, 25 ulcerative colitis and 36 non-IBD patients through therapeutic surgery or endoscopic biopsy. In Crohn disease, 23 surgical specimens and 11 biopsy specimens from initial cases were included. DNA was extracted with phenol-chloroform after homogenization and proteinase K treatment in 73 mucosal specimens. Using an inverted microscope, lymph follicle tissue was microdissected from the remaining 33, mostly biopsy, specimens. DNA was then extracted by freeze thawing. A coagulase gene characteristic of S. aureus was sought. A nested polymerase chain reaction was performed utilizing primers that amplify a region of the coagulase gene. Polymerase chain reaction products were analyzed with polyacrylamide gel electrophoresis. RESULTS: Only one surgically resected colonic specimen, from a 42-year-old male ulcerative colitis patient, registered positive staphylocoagulase amplification. CONCLUSIONS: Staphylococcal superantigens are not involved in either the early lesions or the established lesions of Crohn disease. However, S. aureus infection occasionally may occur during the course of IBD. PMID- 11424321 TI - Impaired sensitivity to beta 2 integrin-blocking in ICAM-1-mediated neutrophil migration in ulcerative colitis. AB - BACKGROUND: Factors influencing the directed migration of neutrophils into colonic tissue in ulcerative colitis (UC) are poorly described. ICAM-1 has recently been shown to possess chemotactic properties, and the aim of this study was to evaluate the involvement of beta 2 integrins in this ICAM-1-mediated migration. METHODS: The chemotactic effect of ICAM-1 on neutrophils isolated from 13 UC patients and 17 healthy volunteers was studied in microchemotaxis chambers. Physiological concentrations of ICAM-1 (0.05-500 pM) were separated from neutrophils by nitrocellulose filters, and cell migration was evaluated using the leading front technique. beta 2 integrins on neutrophils were blocked with antibodies to CD11a, CD11b, CD11c and CD18, and migration towards ICAM-1 was examined. RESULTS: Migration towards ICAM-1 was equal for UC and control neutrophils, showing a bell-shaped ICAM-1 dosemigratory response curve with peak migration at 5 pM ICAM-1 (30.0 microns; interquartile range 22.9-35.7; P < 0.001). Blockade of the CD11 subunits on control cells inhibited the chemoattractant effect of ICAM-1 by 43.6%-58.0%, whereas the migration was decreased by only 20% in UC under similar blocking conditions (P < 0.01). Anti CD18 mAbs had no effect. Inhibition of protein kinases with staurosporin only slightly decreased the ICAM-1-mediated migration, whereas incubation with staurosporin and CD11 antibodies showed additive effects on UC neutrophils and synergistic effects on control cells. No quantitative differences in beta 2 integrin expression were detected between control and UC neutrophils. CONCLUSIONS: The chemotactic property of ICAM-1 was shown to be CD11-dependent and UC neutrophils were found to be less dependent on CD11/ICAM-1-mediated migration than were control neutrophils. PMID- 11424322 TI - Effects of Lactobacillus rhamnosus GG and Lactobacillus reuteri R2LC on acetic acid-induced colitis in rats. AB - BACKGROUND: Certain lactobacilli reduce the severity of experimental colitis. The aim of this study was to compare the effects of a human strain Lactobacillus rhamnosus GG and a rat strain Lactobacillus reuteri R2LC on acetic acid-induced colitis in rats. METHODS: Lactobacillus rhamnosus GG, Lactobacillus reuteri R2LC or sulphasalazine were given orally to the rats. Colitis was assessed 72 h after induction with acetic acid. RESULTS: Lactobacillus reuteri R2LC significantly antagonized body weight loss caused by inflammation compared with Lactobacillus rhamnosus GG and sulphasalazine, and oedema formation in the colon compared with sulphasalazine. Lactobacillus reuteri R2LC reduced the median value of macroscopic ulceration and the protein content of inducible nitric oxide synthase by 50% and the median of the protein content of inducible cyclooxygenase by 30% compared with that of the colitis control group, and Lactobacillus rhamnosus GG reduced the median of inducible nitric oxide protein content by 40% and increased the median of inducible cyclooxygenase protein content by 30% compared with the median value of the colitis control group, but these differences were not statistically significant. CONCLUSIONS: The rat strain Lactobacillus reuteri R2LC, but not the human strain Lactobacillus rhamnosus GG, is of benefit in reducing the severity of acetic acid-induced colitis in rats. These results suggest that it is not the total amount of Lactobacillus but the particular species or strain of Lactobacillus that is important in attenuating experimental colitis. PMID- 11424323 TI - Recombinant human interleukin-6 induces hepatocyte growth factor production in cancer patients. AB - BACKGROUND: Experiments in animals demonstrate an important role for interleukin 6 (IL-6) in liver regeneration. It is suggested that IL-6 initiates hepatocyte growth factor (HGF) synthesis. METHODS: The aim of the study was to examine the effect of exogenously administered recombinant human IL-6 (rhIL-6), in doses of 0.5, 1.0, 2.5, 5, 10 and 20 micrograms/kg/day, on HGF serum levels in humans. Serum HGF levels were measured on days 1, 2, 3, 8 and 15 and were correlated with serum amyloid A (SAA) and C-reactive protein (CRP). RESULTS: Median HGF levels increased to 124% at day 3 (P < 0.05) and 157% (P < 0.05) at day 8 as compared to 100% levels at day 1. An IL-6 dose-dependent increase in HGF was found at day 8 (R = 0.53, P < 0.02). The percentual change in serum HGF level at day 8 correlated with IL-6 serum levels at day 1 R = 0.59, P < 0.01). HGF levels did not correlate with CRP and SAA. CONCLUSION: In humans, rhIL-6 administration resulted in an increase in serum HGF levels. PMID- 11424324 TI - Medical risk factors for small-bowel adenocarcinoma with focus on Crohn disease: a European population-based case-control study. AB - BACKGROUND: Crohn disease and biliary diseases have been associated with small bowel adenocarcinoma (SBA). We examined how medical conditions affect the risk of SBA. METHODS: A population-based European multicentre case-control study during the period 1995-97 including 95 histologically verified cases of SBA along with 3335 population controls; 70 cases (74%) and 2070 (62%) controls were interviewed about previous medical conditions. RESULTS: Crohn disease was identified in two SBA cases (both located in ileum) and two controls; odds ratio (OR) 53.6 (6.0 477) (95% CI in parentheses). Only one case and no controls had had long-standing Crohn disease. Coeliac disease was associated with SBA (2 cases, 0 controls), but one of the cases was diagnosed at the same time as the SBA. Overall, people with a history of gallstones had no increased risk of SBA. The OR was exclusively increased during the 3-year period preceding the SBA diagnosis. Previous gallstone surgery, which may be a sign of severe gallstone disease, was not associated with SBA. Liver cirrhosis, hepatitis or medical treatments with radioactive substances or corticosteroid tablets were not associated with this disease. Cases with SBA had an increased prevalence of anaemia; OR 15.3 (2.5 92.1). An association between low educational level and SBA was found; OR 1.75 (1.0-3.0). CONCLUSION: This study supports Crohn disease and coeliac disease being strong but rare risk factors for SBA. Previous gallstones were unrelated to SBA, and detection bias may account for the findings in earlier studies. PMID- 11424325 TI - Influence of liver fibrosis on hepatic artery Doppler resistance index in chronic hepatitis of viral origin. AB - BACKGROUND: Hepatic arterial Doppler sonography is increasingly being used in liver diagnostics. The determinants of the elevation of hepatic artery impedance indexes in chronic liver disease, however, have still not been fully clarified. The aim of the present study was to investigate the relationship between histological alterations and liver circulation in chronic hepatitis. METHODS: Hepatic artery resistance index and portal flow velocity were measured using Doppler sonography in 47 patients with chronic hepatitis of viral origin diagnosed at histopathology. The patients were divided into two groups, those with mild and those with severe alterations, in accordance with the various histological parameters of the Knodell scoring system. RESULTS: Hepatic artery resistance index and age were higher in patients with more severe liver fibrosis (respectively 0.638 +/- 0.084 and 39.0 +/- 10.9 (years) in mild fibrosis versus 0.687 +/- 0.060 and 49.4 +/- 14.4 (years) in severe fibrosis; P < 0.05 for both), whereas no difference between the two groups was found for the other histological features (degeneration, inflammation and necrosis), nor for portal flow velocity. CONCLUSIONS: The increase in hepatic artery resistance index appears to be influenced by the extent of fibrous tissue deposition in the liver, determined by chronic inflammation and repair and, secondly, by ageing. PMID- 11424326 TI - Effect of beta-adrenergic blockade on elevated arterial compliance and low systemic vascular resistance in cirrhosis. AB - BACKGROUND: Patients with cirrhosis exhibit a characteristic hyperdynamic circulation with increased cardiac output and heart rate and reduced systemic vascular resistance. The compliance of the arterial tree has recently been reported to be increased in these patients, who are often treated with beta blockers, but the effect of this treatment on arterial compliance has not been investigated. The aim of the present study was therefore to assess the effects of propranolol on the arterial compliance of patients with cirrhosis. METHODS: Twenty patients with cirrhosis underwent a haemodynamic investigation with determination of splanchnic and systemic haemodynamics. Arterial compliance was determined as the ratio of the stroke volume to the pulse pressure and compared to normal values. RESULTS: All the patients had significant portal hypertension, with a mean hepatic venous pressure gradient (HVPG) of 17.8 mmHg, and responded to beta-blocker treatment with a significant reduction in the HVPG (-16%; P < 0.001). Arterial compliance was elevated (1.27 versus controls 1.01 ml/mmHg; P < 0.001), but remained almost unchanged during beta-adrenergic blockade (1.27 versus 1.29 ml/mmHg, +2%, ns), whereas systemic vascular resistance increased substantially (1083 versus 1378 dyn x s x cm-5, +27%; P < 0.001). The mean arterial blood pressure (-6%; P < 0.05), heart rate (-20%; P < 0.001), cardiac output (-25%; P < 0.001) and hepatic blood flow (-22%; P < 0.001) fell significantly. CONCLUSIONS: Treatment with beta-blockers increases small vessel (arteriolar) vascular tone towards the normal level, but does not affect the elevated compliance of the larger arteries in patients with cirrhosis. PMID- 11424327 TI - Obesity increases the severity of acute experimental pancreatitis in the rat. AB - BACKGROUND: Studies in patients with acute pancreatitis have shown a correlation between obesity and serious complications or a fatal outcome. However, the mechanisms by which obesity aggravates acute pancreatitis are not known. In the present study we used the sodium taurocholate model of pancreatitis to examine the effect of obesity on severity and outcome in acute experimental pancreatitis (AEP). METHODS: AEP was induced at two degrees of severity by retrograde infusion of sodium taurocholate (0.2 ml x 3% or 0.4 ml x 3.5%) into the pancreatic duct of rats with obesity induced by high-fat diet, genetically obese (GO) rats or lean control rats. Surviving animals were sacrificed 72 h after induction of pancreatitis. RESULTS: In the low-dose experiment, there were no significant differences in pancreatic histology or survival rate between the groups. In the high-dose experiment, the GO rats had a significantly lower 72-h survival rate than the high-fat obese (HFO) or lean control (LC) groups (GO 25% versus HFO 73%, P < 0.05; GO 25% versus LC 100%, P < 0.001). Survival rates in the high-dose experiment correlated strongly with basal liver fat content (R2 = 0.86). Pancreatic histology showed significantly more fat necrosis and a higher total pathological mean score in the HFO rats than in the LC animals (both P < 0.05). CONCLUSIONS: Obesity had a negative influence on the outcome of necrotizing pancreatitis that was related to the magnitude of the pancreatic insult. The sodium taurocholate model in obese rats would be useful for future mechanistic studies of the effect of obesity on pancreatitis. PMID- 11424328 TI - Factors associated with failure of endoscopic injection haemostasis in bleeding peptic ulcers. AB - BACKGROUND: The effectiveness of a submucosal injection of adrenaline solution in endoscopic haemostasis is well documented in patients suffering from peptic ulcer bleeding. After treatment, however, a significant number of patients continue to bleed or rebleed, and require emergency surgical intervention. The aim of this study was to define factors associated with the failure of endoscopic injection haemostatic therapy in peptic ulcer bleeding. METHODS: In the period 1992 to 1998, we prospectively studied all patients suffering from peptic ulcer bleeding and identified endoscopically as being either bleeding actively or carrying a visible vessel. A total of 427 patients (343 men and 84 women; mean age 58.6 +/- 16.6 years) were all subjected to endoscopic injection with adrenaline solution on an emergency basis. Patients who eventually required surgical intervention for permanent haemostasis were considered as endoscopic haemostasis failures, whereas those who did not were considered as endoscopic treatment successes. We evaluated all clinical and endoscopic parameters that might have been related to failure of endoscopic injection therapy. RESULTS: Endoscopic injection haemostasis was successful in 341 patients (79.9%) and a failure in 86 (20.1%) who finally underwent emergency surgical haemostasis. On analysing the examined parameters, failure was significantly related to shock on admission (OR 2.31, 95% CI 1.33, 6.97), spurt bleeding at endoscopy (OR 2.45, 95% CI 1.51, 3.98), posteriorly located duodenal ulcer (OR 2.48, 95% CI 1.37, 7.01) and anastomotic ulcer (OR 3.39, 95% CI 1.37, 7.29). Endoscopic injection haemostasis therapy was less effective in patients with chronic ulcers compared to those who had acute NSAID related ulcers. A history of peptic ulcer (OR 1.57, 95% CI 1.14, 3.05), previous peptic ulcer bleeding (OR 2.45, 95% CI 1.51, 3.98) or non-use of NSAIDs (OR 2.81, 95% CI 1.33, 4.62) were negative predictors for the outcome of endoscopic haemostasis. CONCLUSION: With the use of specific clinical and endoscopic characteristics it is possible to define a subgroup of high-risk patients for continued bleeding or rebleeding despite endoscopic injection therapy. These patients may be candidates for intensive monitoring, early surgical intervention or possibly complementary endoscopic haemostatic methods. PMID- 11424329 TI - Intestinal type of thromboangiitis obliterans (Buerger disease) preceding symptoms of severe peripheral arterial disease. AB - The case of a 50-year-old man with intestinal type thromboangiitis obliterans (Buerger disease) is reported. Intestinal manifestations included stricture and perforation of the colon, and these preceded any symptoms of peripheral vascular disease. Only the histological examination was able to show that our patient had the intestinal type of thromboangiitis obliterans. PMID- 11424330 TI - [Kidney oncocytoma. Report of 7 cases]. AB - Renal oncocytoma has some particularities in diagnosis and prognosis. We report a retrospective study performed between 1987 and 1999 at the department of Urology of Charles Nicolle's Hospital. Seven oncocytoma are found out of 120 renal carcinoma. Features at sonography or CT scan or arteriography can not suggest oncocytoma. The diagnosis is made at the pathology examination. Renal oncocytoma is a benign tumor. Conservative surgery will be performed as possible. PMID- 11424331 TI - [Retroperitoneal ganglioneuroma. Report of a case]. AB - Retroperitoneal ganglioneurom is a primitive, rare and benign tumor originating in the neural sheath. We report the case of a 27-years old man. An abdominal sonography had revealed accidentally a 60 mm, well demarcated tumor. The left renal vein was compressed an forced back. This tumor was located in front of and internal to the left kidney. The CT scan confirmed this tumor. A left subcostal laparotomy was done. The tumor was soft and movable. A total exeresis was performed. The pathology examination had conclude in a benign ganglioneuroma. The follow-up was six years and half. Retroperitoneal ganglioneuroma is a benign tumor with a good prognosis. The treatment is surgical. PMID- 11424332 TI - [Treatment of idiopathic retroperitoneal fibrosis]. AB - Idiopathic retroperitoneal fibrosis is a rare disease and a delay in diagnosis may cause renal failure. Medical treatment i.e. corticotherapy or more recently, tamoxifene has been used successfully. This approach is recommended in patients either with moderate obstruction of the upper urinary, risk of major surgery or in cases of recurrence after surgical treatment. Ureterolysis using conventional surgery or laparoscopy remains the treatment of choice. This procedure should be considered in patients with neoplasic fibrosis, corticoresistant fibrosis or in cases of peri-aneurysm fibrosis. PMID- 11424333 TI - [Retroperitoneal foreign body mimicking pararenal tumor]. AB - We report the case of a 48 year-old woman presently with a pararenal tumor, with a history of pyelotomy in 1978 for the extraction of a renal pelvic stones. Results of surgery showed a foreign body. The clinical and diagnostic aspects of retained surgical gauze have been discussed and the need for radio-opaque markers in them have been emphasized. PMID- 11424334 TI - [Bladder exstrophy in the adult: therapeutic features. Report of 3 cases]. AB - Bladder exstrophy is a malformation which the incidence is estimated at 3.3 per 100,000 births. It is usually treated in the first hours of life. The authors report three cases of bladder exstrophy treated in adult life with continent urinary diversion such as Benchekroun continent ileocaecal bladder (CICB). After a literature review on the management of bladder exstrophy, they studied clinical and therapeutical particularities of the bladder exstrophy in adulthood. PMID- 11424335 TI - [Anatomoclinical study of prostate benign tumors a the urology service of the CHU Ignace Deen]. AB - OBJECTIVE: It was to compare picture and results of anatomopathological exams of the operated organs, given the individualization of new procedures in recent years. METHODS: One hundred and eighty-two patients, aged from 50 to 98 years, were hospitalized for prostatic tumors. After a complete clinical exam followed by surgery, the operated organs were the subject of a conventional histological study. Data from the rectal touch (RT) were compared with the histological results. RESULTS: In the diagnosis of benignancy, RT and the histology are in agreement in 93% of the cases, though only in 75% of the cases of suspected malignancy. Seven percent of the occult cancers are observed, of which ten cases (6%) are adenocarcinoma at Stage A, and two cases (1%) of carcinoma in situ. We observe as well one case of prostatic localization of leukemia and one case of leiomyosarcoma. Prostatitic-type inflammatory lesions and malpighian metaplasias are very frequent 18 and 7%, respectively). Basal cell hyperplasia is rare. CONCLUSION: The RT keeps its place in the exploration of the prostate, within the framework of individual screening and patient follow-up. Transrectal echography and protein-specific antigen (PSA) dosing are certainly necessary but expensive in our country. Histology, which remains irreplaceable, allows us to make a definitive diagnosis at low cost. PMID- 11424336 TI - [Post-traumatic shrinking of the urethra at the urology service of the CHU Ignace Deen: retrospective study on 74 cases]. PMID- 11424337 TI - [Endoscopic internal urethrotomy in the treatment of male urethral stenosis at the urology service of the CHU Ignace Deen]. AB - The authors report a series of 157 stenoses of the urethra treated by endoscopic urethrotomy between 1991 and 1997. After a first urethrotomy the success rate is 51.8%, with a decline of one year. Mortality is zero, and the morbidity assessed at 9%. For these authors, the result is better when the urethrotomy concerns an uninfected, one-time, short (less than 2 cm) stenosis, whatever the etiology, located on the proximal urethra. The duration of the postoperatory catheter has been fixed at three days. Poor results (35.20%) have been reported in stretched stenoses located on the distal urethra. These poor results have been treated by another urethrotomy, with 25% good results. The remaining 10.2% have needed sessions of urethral dilatation, even a plasty. PMID- 11424338 TI - [Surgical treatment of priapism with open glans penis puncture-drainage in 26 black patients]. AB - The surgical treatment of the priapism is complex and requires various techniques for diversion of the corpora cavernosa with significant morbidity. For the last 15 years, the author have been using a simple transglandular cavernosal spongiosum shunt technique, which satisfies the requirements of functional surgery: restoration of anatomy, reduced morbidity, facility to learn. This article is intended to precisely describe the surgical technique and to present the results based on a personal series of 26 cases of priapism operated with this technique in black patients (19 adults and seven children) in the context of sickle-cell anaemia. PMID- 11424339 TI - [Epidemiologic and etiologic features of urinary infections in children at the pediatrics service of the CHU-Campus de Lome (Togo)]. AB - From 1st January 1989 to 31th December 1997, 175 infants (108 females and 67 males) were hospitalised and treated at the pediatric service of CHU-Campus for urinary tract infection; this study follows the observation of the increasing of urinary tract infection in several centers of health in Togo; the aim of this study was to have a list the contributing factors, to understand the mechanism of such infection in order to reduce its frequency and the high percent of the mortality; the diagnosis of urinary tract infection was given by the result of the cytobacteriological exam of the urine which shows the pathological germ; others forms of the investigation, as abdominal echography were used also to look for the etiology of the urinary tract infection; but, the deficit of the of the medical imagery or the old material of the laboratories limited the searching of urinary tract infection etiology; cured infants were declared on the basis of absence of pathological germ in the result of the cytobacteriological exam control of the urine; the prevalence of the urinary tract infection was 8.29% with an incidence of 7.84% at the pediatric service of CHU-Campus; clinics symptoms were atypic and polymorphic; but the fever was the first clinical sign in the newly born and the urological signs were clear only from two to thirty months; 141 children (80.57%) were cured and 34 presented the complications with 3.43% of mortality; preventive measures on the urinary tract infection in infancy were proposed for the children parents and the practical physicians; these measures included information, education and communication (IEC) on the urinary tract infection, the symptomatology and the cytobacteriological exam of the urine. PMID- 11424340 TI - Radiology goes filmless. What does this mean for primary care physicians? AB - Technical advances over the past few years have rapidly increased the possibilities for filmless radiology services provided by PACS. Such systems are also financially feasible for many healthcare practices, primarily because of the goals achieved through increased efficiency and improved patient care. In coming years, most medical facilities are likely to either convert or begin to convert to PACS as conventional film-based systems fade into the annals of medical history. PMID- 11424341 TI - Atopic dermatitis. How to incorporate advances in management. AB - Atopic dermatitis is a common, genetically determined skin disease that most likely results from an immunologic imbalance. In the majority of patients, conservative topical steroid treatment along with patient education suffices. PMID- 11424342 TI - Patient notes: avoiding giardiasis. PMID- 11424343 TI - A new era in opioid dependency treatment. Recent law allows qualified physicians to provide care in office setting. PMID- 11424344 TI - Physician workforce debate continues. PMID- 11424345 TI - Consider obstructive sleep apnea in heart patients. PMID- 11424346 TI - Manifestations of tick-borne illness. Incidence and variety are increasing worldwide. AB - The incidence and variety of diseases associated with tick bites have continued to grow worldwide. Lyme disease, the most common tick-borne disease in the United States, has received extensive media coverage because of its protean manifestations and propensity for causing chronic disease. Our ability to prevent, identify, and effectively treat Lyme disease and other tick-borne diseases has significantly improved in the last decade. Tick-borne illnesses should be one of the differential diagnostic considerations in patients with consistent clinical findings and exposure history. In addition, the prudent use of laboratory testing ensures an accurate diagnosis while avoiding the cost and risk of inappropriate diagnostic tests and antibiotic therapy. PMID- 11424347 TI - How to evaluate an enlarged thyroid? PMID- 11424348 TI - Expanded applications of CT. Helical scanning in five common acute conditions. AB - Helical CT has become a valuable imaging tool for detection of pulmonary embolism, deep venous thrombosis, ureteral colic, acute small-bowel obstruction, and acute appendicitis. Generally, helical CT has good sensitivity and specificity values, and scans can be performed more quickly than previous gold standard diagnostic examinations for the conditions mentioned. In some cases, helical CT can also identify other findings that may be responsible for a patient's symptoms. One notable disadvantage of helical CT is the charge for the procedure, which in some circumstances can be considerably more costly than diagnostic examinations preferred previously. However, because helical CT can often obviate the need for other tests--and may consequently reduce hospital stays--this technology may have the ability to reduce overall expenditures. Cost of helical CT is therefore a multifaceted issue and requires further study before conclusions can be drawn. PMID- 11424349 TI - Nonneurologic indications for MRI. Technological advances have broadened applications. AB - Traditionally used for neurologic indications, magnetic resonance imaging (MRI) is now being used to diagnose the most common musculoskeletal, vascular, hepatic, biliary, adrenal, and renal disorders. Drs Schnitker and Light discuss the nonneurologic indications for MRI as both a primary and an adjunct study. The advantages and disadvantages of fast scanning and open MRI are also presented. PMID- 11424350 TI - Advances in interventional radiology. The search for less invasive management sparks new approaches. AB - Many new, minimally invasive interventional radiology procedures are now viable alternatives to traditional invasive therapy. The radiology procedures can often be performed in the outpatient setting, and although expensive technology may be required, the overall cost to the patient may be lower in the long run. Endovascular repair of abdominal aortic aneurysms with stent grafts and radiofrequency tumor ablation are examples of rapidly expanding technologies in interventional radiology. Patient enthusiasm and interest are the primary forces driving these advances in management. Familiarity with these procedures is vital as medicine moves into the new millennium. PMID- 11424351 TI - [Phenotype--genotype relationships in peroxisome biogenesis disorders]. PMID- 11424352 TI - [Mitochondrial dysfunction and brain development disorders]. PMID- 11424353 TI - [Menkes' disease and brain dysfunction]. PMID- 11424354 TI - [Brain abnormalities in genomic imprinting diseases]. PMID- 11424355 TI - [Current status of treatment of brain tumors in Japan: from the report of brain tumor registry of Japan]. PMID- 11424356 TI - [Innervation pattern to the extensor digitorum brevis by deep peroneal nerve and accessory deep peroneal nerve]. AB - On the electrophysiological basis, extensor digitorum brevis(EDB) muscle is innervated electrophysiologically not only by deep peroneal nerve(DPN) but also by accessory deep peroneal nerve(ADPN), an anomalous branch of superficial peroneal nerve, with a prevalence of 17-28%. We investigated 23 patients who had both DPN and sufficient ADPN innervation to the EDB on the intramuscular distribution of DPN and ADPN innervation to the medial and lateral side of the EDB. Recording electrodes were placed on the medial and lateral edges of the EDB with a supramaximal stimulation to the anterior or lateral ankle, compound muscle action potential (CMAP) of DPN or ADPN innervation was recorded. In 19 patients (83%) the DPN innervation was larger than or equal to the ADPN innervation. Only in 4 patients (17%) the ADPN innervation obviously exceeded the DPN innervation. DPN enters to the EDB from the medial side, and ADPN from the lateral side of the EDB. In 16 patients(70%) the DPN innervation was relatively large and the ADPN innervation was relatively small at the medial side of the EDB, and vice versa at the lateral side of the EDB. These distributions were almost uniform in 5 patients(22%). This study clarified that a biased larger DPN innervation and smaller ADPN innervation to the medial side of the EDB, and vice versa to the lateral side of the EDB in the majority cases. In some cases diffuse innervation to the EDB was found. PMID- 11424357 TI - [Minimally invasive stereotactic functional surgery using an intravenous anesthetic propofol and applying Image Fusion and AtlasPlan]. AB - Image-guided stereotactic surgery of the ventralis intermedius nucleus of the thalamus, globus pallidus, and subthalamic nucleus is a prevailing modality as a treatment of movement disorders. This technical note describes a method of minimally-invasive stereotactic functional surgery for patients with parkinsonian symptom or various tremors. Patients were administered propofol, an intravenous anesthetic, during placement of a burr hole and a stereotactic frame, but not the period when the patients were necessary to be awake. The intravenous anesthesia was very beneficial to relieve local pain and mental stress of the patients. Radionics ImageFusion and AtlasPlan were used for defining the target localization without an intraoperative ventriculography. ImageFusion efficiently fused high-resolution MR images on CT images. AtlasPlan accurately corrected the localization of the tentative target point after measurements from the midpoint of the anterior commissure-posterior commissure line on the modified MRI, and enabled us to reduce the degree of an intraoperative correction to fix a final target. Stereotactic functional surgery has been thought to be less-invasive, however further modifications of surgical procedure and intraoperative medication can make this type of surgery much more less-invasive. PMID- 11424358 TI - [Usefulness of time-resolved projection MRA on evaluation of hemodynamics in cerebral occlusive diseases]. AB - The usefulness for evaluation of cerebral hemodynamics using time-resolved projection MRA was studied in normal volunteers and patients of cerebrovascular diseases. Six normal volunteers and ten patients with cerebrovascular occlusive diseases including 6 of IC occlusion and 4 of post EC/IC bypass surgery underwent time-resolved projection MRA on a 1.5 T clinical MRI system. Projection angiograms are acquired with 2 D-fast SPGR sequence with a time resolution of approximately one image per second, 40 images being acquired consecutively before and after bolus injection Gd-DTPA. And all images were calculated by complex subtraction from the background mask in a work station. In normal volunteers, the quality of images of time-resolved projection MRA was satisfactory. The arteries from internal carotid artery through M 2 segment of middle cerebral artery and all major venous systems were well portrayed. In 4 cases of IC occlusion who were assessed the collateral flow through the anterior communicating artery and posterior communicating artery, there were delayed to demonstrate the ipsilateral MCA. However, in 2 cases of IC occlusion that were assessed the collateral flow through leptomeningeal anastomosis, ipsilateral MCA and collateral circulation were not demonstrated. In all patients of post EC/IC bypass surgery, the patency of EC/IC bypass could be evaluated as properly with time-resolved projection MRA as 3 D-TOF MRA. Although the temporal and spatial resolutions are insufficient, time-resolved projection MRA was powerful non-invasive method to evaluate the cerebral hemodynamics via the basal communicating arteries in IC occlusion and identify the patency of EC/IC bypass. PMID- 11424359 TI - [Multicystic sphenoethmoidal mucocele with intracranial extension presented with generalized convulsion: a case report]. AB - A 53-year-old male presented with generalized convulsion. Magnetic resonance imaging and computed tomography revealed large sphenoethmoidal mucocele extending intracranially. Signal intensities between paranasal and intracranial portion of the mucocele were different. Preoperatively it was not obvious that intracranial portion of the mucocele communicated with paranasal portion of the mucocele. We approached the paranasal portion of the mucocele by transsphenoidal approach resulting in disappearance of the intracranial portion of the mucocele. Sphenoethmoidal mucocele with intracranial extension presenting with generalized convulsion was the first case in the literature. Transsphenoidal approach for sphenoethmoidal mucocele with intracranial extension is the first choice of surgical treatment, though neuroimaging shows that there is no communication between paranasal and intracranial portion of mucocele. PMID- 11424360 TI - [An adult case of cerebellar mutism after removal of cerebellar hematoma]. AB - Cerebellar mutism is a well documented complication of posterior fossa tumor surgery in children. But only one case presenting with cerebellar mutism after removal of a cerebellar hematoma in adults was reported previously. We present 64 year-old male developing cerebellar mustism after surgery for hypertensive cerebellar hemorrhage. A cerebellar hematoma was located in the right side dentate nucleus and cerebellar peduncle dominantly not extending to the cerebellar vermis. During the period of cerebellar mutism, single photon emission computed tomography revealed a decrease in the contralateral temporo-parietal cerebral blood flow. Cerebellar mutism in this case may be associated with the low blood flow in the contralateral supplementary motor cortex owing to injury to the dentate-thalamo-cortical pathway at the dentate nucleus and cerebellar peduncle. The relevant literature is reviewed. PMID- 11424361 TI - [Surgical management for an unruptured cerebral aneurysm accompanied by anti phospholipid antibody syndrome: a case report]. AB - Anti-phospholipid antibody syndrome(APS) is characterized by the presence of antibodies to phospholipids and various symptoms including those derived from thrombosis. APS has been shown to be frequently accompanied by multiple and recurrent cerebral ischemic lesions, suggesting that APS may play a critical role in developing cerebral ischemia of unknown etiology. Here, we present a case with an unruptured cerebral aneurysm accompanied by a severe stenosis of the middle cerebral artery as a manifestation of APS. The aneurysm was successfully treated surgically employing the following precaution to prevent potential damage on the fragile ischemic brain: preoperative administration of anticonvulsant, intermittent brain retraction, intraoperative SEP monitoring, and postoperative administration of anticoagulant. Our experience suggests that unruptured cerebral aneurysms accompanied by APS could be safely treated by strict paraoperative management. PMID- 11424362 TI - [Nerve hypertrophy in a patient with chronic inflammatory demyelinating polyneuropathy]. PMID- 11424363 TI - [Small cortical infarction presenting isolated sensory disturbance on right upper limb: diagnosis by diffusion-weighted MRI]. PMID- 11424364 TI - [Multiple brain embolisms with bilateral sudden deafness at the onset]. PMID- 11424365 TI - [The role of CD44 in the invasion and metastasis of gastric cancer]. PMID- 11424366 TI - [Matrix metalloproteinase in human gastric carcinoma]. PMID- 11424367 TI - [Significance of gastritis on the gastric carcinogenesis]. PMID- 11424368 TI - [Histologic classification of gastric carcinoma from the view-point of its histogenesis]. PMID- 11424369 TI - [Risk factors for stomach cancer]. PMID- 11424370 TI - [Pathological feature of the growth of gastric cancer]. PMID- 11424371 TI - [Clinicopathological findings and histological appearance of early gastric cancer]. PMID- 11424372 TI - [Japanese Classification of Gastric Carcinoma (the 13th edition, June 1999): points to be revised]. PMID- 11424373 TI - [Radiological diagnosis of gastric cancer, current trends]. PMID- 11424374 TI - [Electronic endoscopy for diagnosis of gastric cancer]. PMID- 11424375 TI - [Endoscopic ultrasonography for the diagnosis of gastric cancer]. PMID- 11424376 TI - [CT and MR diagnosis of metastatic lesions in patients with gastric cancer]. PMID- 11424377 TI - [Diagnosis of the lymphatic metastasis of gastric cancer by sentinel node biopsy]. PMID- 11424378 TI - [Serum pepsinogen test for the diagnosis of stomach cancer]. PMID- 11424379 TI - [Tumor marker in gastric cancer]. PMID- 11424380 TI - [Gene diagnosis of gastric cancer]. PMID- 11424381 TI - [Histopathological diagnosis of gastiric cancers]. PMID- 11424382 TI - [Clinical study of early gastric cancer]. PMID- 11424383 TI - [Advanced gastric cancer]. PMID- 11424384 TI - [Recurrence of gastric cancer]. PMID- 11424385 TI - [Epidemiology of stomach cancer and Helicobacter pylori infection]. PMID- 11424386 TI - [Multiple gastric cancer]. PMID- 11424387 TI - [Gastric cancer of the stomach remnant]. PMID- 11424388 TI - [Familial gastric cancer]. PMID- 11424389 TI - [Scirrhous carcinoma of the stomach--from bench to clinic]. PMID- 11424390 TI - [Gastric MALT lymphoma]. PMID- 11424391 TI - [Myogenic tumor of the stomach]. PMID- 11424392 TI - [Surgical treatment for gastric cancer in western countries]. PMID- 11424393 TI - [Establishment of clinical practice guidelines for gastric cancer and its significance]. PMID- 11424394 TI - [Surgical indication for early gastric cancer]. PMID- 11424395 TI - [Endoscopic mucosal resection]. PMID- 11424396 TI - [Laparoscopic surgery for early gastric cancer]. PMID- 11424397 TI - [Limited surgery for early gastric cancer]. PMID- 11424398 TI - [Molecular mechanisms of carcinogenesis and progression of human stomach cancer]. PMID- 11424399 TI - [Laser endoscopy for therapy of early-stage gastric cancer]. PMID- 11424400 TI - [Endoscopic microwave coagulation therapy for early gastric cancer]. PMID- 11424401 TI - [Management of postoperative complications (functional disorder of gastro intestinal tract, infection, vitamin deficiency, metabolic disorder of bone)]. PMID- 11424402 TI - [Standard surgical treatment of gastric cancer]. PMID- 11424403 TI - [Extended surgery for advanced gastric cancer]. PMID- 11424404 TI - [Decision making: how to choice anticancer chemotherapy]. PMID- 11424405 TI - [Standard chemotherapy for gastric cancer]. PMID- 11424406 TI - [Response criteria of chemotherapy for gastric cancer]. PMID- 11424407 TI - [Low-dose administration of 5-fluorouracil and cisplatin for advanced gastric cancer]. PMID- 11424408 TI - [MTX/5FU sequential therapy for advanced and recurrent gastric cancer]. PMID- 11424409 TI - [Chemotherapy of irinotecan (CPT-11) combined with cisplatin in patients with advanced gastric cancer]. PMID- 11424410 TI - [Combination therapy of eucovorin and 5-FU for advanced gastric cancer]. PMID- 11424411 TI - [Irinotecan (CPT-11) therapy for advanced gastric cancer]. PMID- 11424412 TI - [Genetic alteration in gastric cancer]. PMID- 11424413 TI - [TS-1 therapy for advanced gastric cancer]. PMID- 11424414 TI - [Capecitabine (Ro09-1978) for therapy of advanced and recurrent gastric cancer]. PMID- 11424415 TI - [New drugs for unresectable or recurrent gastric cancer--paclitaxel and docetaxel]. PMID- 11424416 TI - [Management of anticancer drug adverse reactions]. PMID- 11424417 TI - [Chemosensitivity test for advanced and recurrent gastric cancer]. PMID- 11424418 TI - [Immunochemotherapy for cancer patients of the stomach--cancer therapy keeping with good host-immunocompetency]. PMID- 11424419 TI - [CDK inhibitors for therapy of cancer]. PMID- 11424420 TI - [Matrix metalloproteinase inhibitors for therapy of cancer]. PMID- 11424421 TI - [Cancer vaccine with peptides derived from tumor rejection antigens]. PMID- 11424422 TI - [Molecular mechanism of carcinogenesis in human stomach cancer: Bub1 gene]. PMID- 11424423 TI - [Gene therapy for gastric cancer]. PMID- 11424424 TI - [Adjuvant chemotherapy for gastric cancer]. PMID- 11424425 TI - [Neoadjuvant chemotherapy for advanced gastric cancer]. PMID- 11424426 TI - [Characteristics of tumor microcirculation and selective enhancement of drug delivery--angiotensin II induced hypertension chemotherapy for gastric carcinoma]. PMID- 11424427 TI - [Chemo-hyperthermic peritoneal perfusion for therapy of advanced gastric cancer with peritoneal dissemination]. PMID- 11424428 TI - [Palliative medicine and palliative care of cancer]. PMID- 11424429 TI - [Molecular mechanism of carcinogenesis in human stomach cancer: Genetic polymorphism of cytochrome P4502E1]. PMID- 11424430 TI - [The treatment of gastric cancer with distant metastasis]. PMID- 11424431 TI - [Prevention of stomach cancer from a view point of epidemiologic trends]. PMID- 11424432 TI - [Lifestyle factors and the prevention of stomach cancer]. PMID- 11424433 TI - [Epidemiologic trends of stomach cancer in Japan and world]. PMID- 11424434 TI - [Gastric cancer prevention by Helicobacter pylori eradication]. PMID- 11424435 TI - [Prevention of gastric stump carcinoma]. PMID- 11424436 TI - [The latest tumor marker in gastric cancer]. PMID- 11424437 TI - [New molecular markers for prognosis of gastric cancer]. PMID- 11424438 TI - [Factors influencing prognosis of gastric cancer: tumor characteristics and therapeutical aspects]. PMID- 11424439 TI - [Molecular mechanisms of carcinogenesis in human stomach cancer: K-sam gene]. PMID- 11424440 TI - [Effectiveness and cost-benefit of screening for gastric cancer in Japan]. PMID- 11424441 TI - [Evaluation of serum pepsinogen test for screening of gastric cancer]. PMID- 11424442 TI - [QOL assessment of two randomized controlled trials of adjuvant chemotherapy after gastric resection--an interim report of JFMC]. PMID- 11424443 TI - [Informed consent for surgical treatment of gastric cancer]. PMID- 11424444 TI - [Informed consent in gastric cancer patients treated with chemotherapy]. PMID- 11424445 TI - [Multidisciplinary support of cancer patients for truth telling]. PMID- 11424446 TI - [Natural history of gastric cancer]. PMID- 11424447 TI - [Personalized chemotherapy for gastric cancer: new chemotherapeutic strategy]. PMID- 11424448 TI - [The expression of PPAR gamma in gastric cancer]. PMID- 11424449 TI - [COX-2 expression in gastric cancers]. PMID- 11424450 TI - [Role of Epstein-Barr virus in the oncogenesis of gastric carcinoma]. PMID- 11424451 TI - [Perioperative risk factors and its evaluation in the operation for gastric cancer of aged patients]. PMID- 11424452 TI - [Cancer chemotherapy for gastric cancer patients in the elderly]. PMID- 11424453 TI - [Home chemotherapy and/or outpatient chemotherapy for patient with advanced gastric cancer]. PMID- 11424454 TI - [Endosonography-guided celiac plexus neurolysis for cancer pain]. PMID- 11424455 TI - [Molecular mechanisms for Helicobacter-related gastric carcinogenesis]. PMID- 11424456 TI - [Glandular stomach carcinogenesis in animal models infected with Helicobacter pylori]. PMID- 11424457 TI - [Virulence factors of Helicobacter pylori and gastric cancer]. PMID- 11424458 TI - [Molecular mechanisms of early invasion of gastric cancer cells]. PMID- 11424459 TI - [Molecular mechanism of carcinogenesis in gastric cancer: Intercellular molecule cadherin and catenin]. PMID- 11424460 TI - [The role of integrin in the invasion and metastasis of gastric cancer]. PMID- 11424461 TI - [Effectiveness of Saiko-Keishi-To (TJ-10, a Kampo herbal medicine) for trigeminal neuralgia in rats]. AB - It is known that Saiko-Keishi-To (TJ-10), a Kampo herbal medicine used for the treatment of epilepsy, shows a satisfactory curative effect even in the patients suffering from trigeminal neuralgia. To verify the effectiveness of TJ-10, Wistar rats with chronic neuralgia of the mandibular nerve were prepared and TJ-10 was administered to them for 4 weeks following the manifestation of pain in the mandibular region. The result reveals that the rise in the pain threshold in the mandibular region is more significant in the rats administered TJ-10 than in those in the control group. However, in the tail flick test, no significant change was observed in the pain threshold. These findings suggest that TJ-10 is effective for controlling the manifestation of pain in ligatured nerves, by local effect, not by general analgesic effect. PMID- 11424462 TI - [The effects of conscious sedation by propofol on respiration during abdominal hysterectomy under spinal anesthesia]. AB - The effects of conscious sedation by propofol on respiration were studied in 28 patients undergoing abdominal hysterectomy under spinal anesthesia. The patients were randomly assigned to receive conscious sedation by propofol (Group P, n = 20) or no sedation (Group C, n = 8). After a satisfactory level of analgesia had been achieved, a loading dose of propofol, 0.2 mg.kg-1 was administered every minute in Group P until patients exhibited spontaneous eye closure or nystagmus. Thereafter, the infusion rates were adjusted to maintain conscious sedation. Respiratory rate, SpO2 and nasal PETCO2 were measured every minute during the initial 30 minutes and subsequently at 5-min intervals. The patients were questioned on the 1st postoperative day concerning intraoperative recall and the level of satisfaction. The mean loading dose of propofol was 0.4 mg.kg-1 and the mean mainteinance rate was 1.7 +/- 0.5 mg.kg-1.h-1. Compared with Group C, the respiratory depression was less in Group P; SpO2 was significantly higher at 25 min after spinal tap and PETCO2 was significantly lower at 30 and 50 min after spinal tap in Group P. A score of patient satisfaction was significantly higher in Group P. Conscious sedation by propofol is a safe and useful supplement to spinal anesthesia for abdominal hysterectomy. PMID- 11424463 TI - [The techniques to identify the epidural space do not influence the success rate in combined spinal-epidural anesthesia: a comparison between loss-of-resistance and hanging-drop]. AB - The hanging-drop (HD) technique has been attributed to a negative epidural pressure induced by making a tent of the dura by the Tuohy needle. We, therefore, hypothesized that the HD technique would result in more successful intrathecal placement of the spinal needle in combined spinal-epidural anesthesia (CSEA) compared with the loss-of-resistance (LR). Seventy patients received CSEA using the needle-through-needle method with a spinal needle extending 9 mm beyond the Tuohy needle. Success rate in obtaining cerebrospinal fluid (CSF) at the first attempt was considerably low; 60% in HD and 57% in LR, which was not significantly different. Failure to obtain CSF after 3 attempts was not significantly different between the two techniques; 26% and 31% in HD and LR, respectively. In conclusion, there was no advantage of the HD technique for obtaining CSF in CESA compared with the LR. It seems that spinal needle length beyond the Tuohy needle should be more than 9 mm. PMID- 11424464 TI - [Propofol Concentration during Maintenance of and Emergence from Propofol/Epidural Anesthesia: Comparison between Younger and Elderly Patient]. AB - To investigate the influence of patient age on the sensitivity to propofol, we measured blood propofol concentrations in ten elderly (over 70 years of age) and ten younger (under 60 years of age) patients undergoing elective abdominal surgery during propofol/epidural anesthesia. Bispectral index (BIS) was continuously recorded for monitoring anesthetic effect, and the infusion rate of propofol was controlled to keep BIS at 50 after the induction of anesthesia with propofol. At steady-state before skin incision, propofol concentrations to maintain BIS at 50 in younger and elderly patients were 4.3 +/- 1.6 micrograms.ml 1 and 3.2 +/- 1.3 micrograms.ml-1, respectively, and there was no significant difference. Mean blood propofol concentrations were 4.0 +/- 1.5 micrograms.ml-1 for the younger group and 3.2 +/- 1.7 micrograms.ml-1 for the elderly group at the end of surgery immediately before discontinuation of propofol, and they were 1.9 +/- 0.7 micrograms.ml-1 (BIS = 86 +/- 7) for the younger group and 1.5 +/- 0.8 micrograms.ml-1 (BIS = 84 +/- 3) for the elderly group at the time of eye opening with no significant difference. We found appreciable individual variations in the propofol concentrations both in younger and elderly patients in this study. PMID- 11424465 TI - [Appearance of angry backfiring C-nociceptor (ABC) syndrome-like pain disorder in cross-side of one-side spinal anesthesia]. AB - ABC (angry backfiring C-nociceptor) syndrome-like pain disorder categorized in sympathetic independent pain appeared after amputation during spinal anesthesia. A 69-year-old female who had undergone amputation of the right leg below knee 5 months before, received amputation of the left leg below knee this time because of diabetic neuropathy and gangrene of her both legs. There were no complications in her right leg after the former amputation. Spinal anesthesia was maintained in the left one-side by hyperbaric local anesthetic. After changing the body position from left lateral to supine, the left leg was completely anesthetized and the right leg was incompletely anesthetized with only loss of cold sensation. In this state, she had pain in her right leg, and this pain was relieved by intravenous lidocaine administration. It was indicated that this type of pain was caused by the presence of ABC syndrome probably after right leg amputation. PMID- 11424466 TI - [Severe pulmonary edema in a preeclamptic patient with peripartum cardiomyopathy]. AB - We report a preeclamptic patient who revealed severe pulmonary edema. A 37-year old woman, at 32-week gestation, underwent emergency cesarean section under general anesthesia. She had no particular past medical history. SpO2 was 84% (room air) on the arrival at the operating room, and a chest X-ray showed pulmonary edema. She was managed in the intensive care unit postoperatively, she received an intensive treatment (positive pressure ventilation, furosemide, and inotropic support), and was extubated successfully on the third postoperative day. From clinical course and echocardiographic findings, we consider that this pulmonary edema was caused by peripartum cardiomyopathy. PMID- 11424467 TI - [Relationship between minimum alveolar concentration and electroencephalographic bispectral index as well as spectral edge frequency 95 during isoflurane/epidural or sevoflurane/epidural anesthesia]. AB - To investigate the relationship between minimum alveolar concentration (MAC) and electroencephalographic variables, we measured the bispectral index (BIS) and the spectral edge frequency 95 (SEF 95) in 17 patients undergoing elective surgery during isoflurane/epidural (n = 8) or sevoflurane/epidural (n = 9) anesthesia. Patients received 2.0 MAC end-tidal concentrations of isoflurane or sevoflurane, and the BIS and the SEF 95 were recorded after 15 min of an unchanged end-tidal concentration. The concentration of the inhalational agent was decreased to 1.2 MAC, and measurements were repeated again. During isoflurane anesthesia, the BIS increased significantly (3.6 +/- 3.9 at 2.0 MAC, 43.5 +/- 9.2 at 1.2 MAC [mean +/ SD]). In contrast, the BIS did not change significantly during sevoflurane anesthesia (35.3 +/- 8.4 at 2.0 MAC, 42.8 +/- 6.1 at 1.2 MAC). There were significant differences in the BIS and the SEF 95 at 2.0 MAC between isoflurane and sevoflurane groups. In contrast, the BIS and the SEF 95 showed no difference at 1.2 MAC between the groups. These findings suggest that different inhalational anesthetics may have different effects on the BIS and the SEF 95. PMID- 11424468 TI - [Intubation with a double lumen tube in patients with difficult airway using endoscopy mask, and tracheal tube guide]. AB - A 71-year-old-man with pneumothorax was scheduled for a video-assisted-thoracic surgery. He had received subtotal resection of the mandible and was anticipated to have difficult airway. Anesthesia was induced by fentanyl, midazolam, and maintained by 0.4% isoflurane in pure oxygen. The patient was ventilated using Endoscopy Mask, having a port for fiberscopy. A fiberoptic bronchoscope and an endotracheal tube can be passed through the port without significant air leakage during positive pressure ventilation. First, a single lumen orotracheal tube was passed through the port, and Endoscopy Mask was removed. Then, a Tracheal Tube Guide was inserted through the single lumen tube. After the tube had been removed, a double lumen tube was advanced into the trachea guided by the Tracheal Tube Guide. During this procedure oxygen saturation was maintained at 100%. PMID- 11424469 TI - [Orotracheal intubation using a styletscope in a patient to avoid neck recurvation]. AB - We used a Styletscope for endotracheal intubation in a 71-year-old man in whom neck recurvation is contraindicated. Orotracheal intubation was accomplished promptly with the scope. We conclude that Styletscope is useful for orotracheal intubation in patients with limited neck recurvation. PMID- 11424470 TI - [Myasthenia gravis is improved temporarily at postburn period]. AB - A 62-year-old woman with a 25-year history of myasthenia gravis (MG) was admitted to our hospital due to burn injury over 20-25% of the total body surface area. Five months before admission, the serum concentration of acetylcholine receptor (AchR) antibodies was 80.9 nmol.l-1 (normal range < 0.3 nmol.l-1). Anticholinesterase agents had been administered for MG, but were discontinued six days after admission due to muscarinic side effects, but no symptoms of MG appeared. Thirteen days after admission, the AchR antibody titer was 21.2 nmol.l 1. Free skin grafting was performed under general anesthesia without any event. About 80 days after admission, weakness of extraocular muscles appeared. Positive tensilon test and the characteristic electromyographic findings revealed deterioration of MG, and anticholinesterase agents were resumed. Ten months after admission, the AchR antibody titer was 50.4 nmol.l-1. The mechanism of the temporary improvement of MG symptoms does not appear to be explained by the diffuse immunosuppression after burn. At a postburn period, nicotinic AchRs at the neuromuscular junction are known to be temporarily induced. This up regulation may have caused the temporary improvement in this patient. PMID- 11424471 TI - [A case of pulmonary edema after electroconvulsive therapy under propofol anesthesia]. AB - Electroconvulsive therapy (ECT) was scheduled for a 61-yr-old woman with major depression who had been taking a beta-blocker for hypertension. She underwent the first ECT under thiamylal anesthesia uneventfully. The second ECT was performed under propofol anesthesia on the next day. Immediately after ECT, the heart rate dropped from 56 to 19 beats.min-1, which was remedied by intravenous atropine. Then, the blood pressure increased to 204/108 mmHg but it was controlled by nicardipine. However, the SpO2 decreased to 84-88% under oxygen administration by mask at a rate of 3 l.min-1. The patient complained of chest discomfort and had a bloody secretion from the trachea. A chest X-ray showed a butterfly shadow. The patient was diagnosed as having neurogenic pulmonary edema and was treated in the ICU by artificial ventilation and administration of diuretics and catecholamines. These treatments proved to be successful, and the patient was discharged from the ICU 4 days later uneventfully. This case indicates that hemodynamics should be carefully monitored following ECT and that care should be taken to prevent the occurrence of complications after ECT. PMID- 11424472 TI - [Anesthetic management of a hyper-obese patient by target-controlled infusion (TCI) of propofol and fentanyl]. AB - We gave total intravenous anesthesia to an over-100% hyper-obese patient using target-controlled infusion (TCI) of propofol and fentanyl. To keep him asleep, we maintained his BIS in a range of 40 to 60 by adjusting the target concentration of propofol. For the target concentration of fentanyl, we chose 2 ng.ml-1 at incision and 1.6 ng.ml-1 during the operation. At the patient's emergence from anesthesia, his estimated blood concentration of propofol was 1.51 micrograms.ml 1 and his BIS was 80. The relationship between BIS value and effect-site concentration of propofol was almost the same as that assessed in ordinary adults of a normal weight. We conclude that the estimated concentration of propofol is a good indicator of the effect of propofol and that TCI is a useful technique in obese patients as well as in ordinary adults. PMID- 11424473 TI - [Anesthetic management of a patient with tetanus using epidural anesthesia]. AB - A 75-year-old woman with breast cancer complicated with tetanus was scheduled for mastectomy. Since severe bradycardia (17 beats.min-1) was detected by preoperative Holter monitoring, a temporary pacing catheter was inserted. She underwent mastectomy under general anesthesia using propofol combined with thoracic epidural anesthesia. She also received postoperative thoracic epidural block. Her perioperative heart rate was 80-105 beats.min-1 and the rhythm was sinus. There was no marked perioperative cardiovascular derangement. PMID- 11424474 TI - [Anesthetic management of a patient with blood coagulation factor XII deficiency]. AB - A 36-yr-old pregnant woman with blood coagulation factor XII deficiency was scheduled to undergo cesarean section. Preoperative blood coagulation scan revealed prolonged activated partial thrombin time of 105.6 s (control, 30.3 s), and a decrease in factor XII level of 3% (normal; 70-120%). Other examinations such as prothrombin time, platelet count and bleeding time were within normal limits. But she reported no history of bleeding. Before the operation, we discussed with obstetricians whether fresh frozen plasma (FFP) should be administered, or not. According to a report, a patient with factor XII deficiency can undergo major surgery without plasma replacement therapy. But another study showed that 15-30% of the factor XII activity was needed for hemostasis. We, therefore, administered 4 units of fresh frozen plasma before the operation. The operation was performed with general anesthesia uneventfully. Generally, a patient with factor XII deficiency has no clinical bleeding problems and unnecessary FFP administration may cause infection. Therefore it is not necessary to administer FFP to this patient. PMID- 11424475 TI - [Successful management of a patient who developed intra-operative pulmonary tumor embolism]. AB - A 68-year-old female with retroperitoneal tumor extending into the inferior vena cava (IVC) developed massive pulmonary tumor embolism during removal of the tumor. Because of her unstable hemodynamics, emergency pulmonary embolectomy under cardiopulmonary bypass was performed. Successful management of her intra- and post-operative persistent right heart failure led to a satisfactory postoperative course without serious neurological complications. In peri operative management of a patient with an extended tumor into IVC, prevention of the embolism, detection of the pulmonary embolism and treatment of intra- and post-operative right heart failure are important. PMID- 11424476 TI - [Anesthesia for patients with unexpected pulmonary embolisms due to a large abdominal tumor obstructing with inferior vena cava]. AB - Several reports have highlighted the risk of pulmonary embolisms during the preoperative period in patients with large abdominal tumors obstructing the inferior vena cava. We describe a patient who developed pulmonary embolism just before surgery. A 68[correction of 64]-year-old female was scheduled to undergo elective resection of a large abdominal tumor under general anesthesia. She had no signs of deep venous thrombosis, but on the day of the operation, pulmonary embolism developed suddenly. Anticoagulant therapy was performed. Capnography and pulmonary artery pressure were monitored during the perioperative period to detect the recurrence of pulmonary embolism. The percutaneous cardio-pulmonary support (PCPS) was also prepared. The operation was performed successfully. In this patient, the pulmonary embolism occurred suddenly during the preoperative period, even though we had ruled out the existence of deep venous thrombosis. This case report emphasizes the risk of pulmonary embolism in any patient with a large abdominal tumor obstructing the inferior vena cava. This case of a large abdominal tumor suggests that capnography, monitoring of pulmonary artery pressure and preparation of a PCPS in case of pulmonary embolism during surgery are necessary even in patients without signs of deep venous thrombosis. PMID- 11424477 TI - [Anesthetic management of two patients with essential thrombocythemia]. AB - We report different methods of anesthetic management in two patients with essential thrombocythemia. Case 1 is a 69-year-old male scheduled for cholecystectomy. His blood platelet counts were maintained between 10 to 40 x 10(4).microliters-1 after myelosuppression therapy. His preoperative blood tests were within normal limits. Since he had no signs of hemorrhage or thrombus preoperatively, an epidural catheter was inserted for intraoperative analgesia and postoperative pain relief. Anesthesia was induced with propofol and fentanyl, and maintained with N2O-O2-sevoflurane. Mepivacaine 1% was injected through the epidural catheter for intraoperative analgesia and buprenorphine was injected through the catheter for postoperative pain relief. His perioperative course was uneventful. Case 2 is an 88-year-old female scheduled for emergency enterectomy. She had had recurrent bouts of thrombosis. Her blood platelet counts were 89.1 x 10(4).microliters-1. Since her preoperative management of thrombocythemia had been poor, epidural anesthesia was not performed. Anesthesia was induced with propofol, and maintained with N2O-O2-sevoflurane. Her perioperative course was uneventful. We conclude that spinal or epidural anesthesia is not contraindicated when preoperative platelet counts and aggregation test are within normal limits in a patient with essential thrombocythemia. PMID- 11424478 TI - [Treatment with stellate ganglion block, continuous epidural block and ulnar nerve block of a patient with postherpetic neuralgia who developed complex regional pain syndrome (CRPS)]. AB - We present a case of a 46-year-old female patient with systemic lupus erythematosus who developed herpes zoster of the right eighth cervical nerve. Her whole right forearm, hand and the first through fifth fingers were coated with some gel and protected against pain. She had been suffering from continuous and spasmodic burning pain, hyperalgesia, allodynia, drop in skin temperature, sudmotor disturbance, edema, constructure of the joints, muscle atrophy and bone atrophy of her right upper extremity probably due to postherpetic neuralgia (PHN) associated with complex regional pain syndrome (CRPS). She received right stellate ganglion block (SGB), continuous cervical epidural block and right ulnar nerve block. Reduction of pain and edema as well as improvement in mobility of each joint of her right upper extremity was observed. We suspect that SGB, continuous cervical epidural block and ulnar nerve block are effective and useful alternative treatments in a patient with PHN associated with CRPS of the eighth cervical nerve. PMID- 11424479 TI - [Anesthetic management of a patient with Crow-Fukase syndrome complicated with severe heart failure]. AB - A 31-year-old man with Crow-Fukase syndrome was scheduled for orthopedic surgery of the right lower limb. Preoperative examination revealed severe heart failure due to cardiomyopathy. Anesthesia was maintained with sevoflurane and nitrous oxide in oxygen, and supplemental fentanyl combined with continuous epidural block. During surgery, we evaluated cardiac function using transesophageal echocardiography. To maintain his cardiac function, intravenous dopamine and dobtamine were administered continuously, and the surgery was performed successfully. We must take care of cardiac function during anesthetic management of a patient with this syndrome. PMID- 11424480 TI - [Anesthesia for thoracotomy in a patient with severe emphysema associated with Wolff-Parkinson-White syndrome]. AB - A 57-yr-old man with Wolff-Parkinson-White syndrome was scheduled for thoracotomy due to pneumothorax caused by severe emphysema (FEV1.0% 29%). Anesthesia was induced with propofol and fentanyl and maintained with continuous propofol infusion combined with thoracic epidural anesthesia. During mechanical ventilation, the peak inspiratory pressure was reduced to avoid overinflation or rupture of the lung. Although severe hypercapnia was observed during one lung ventilation, there was no incidence of tachyarrhythmias that we had feared. We suggest that hypercapnia is unlikely to cause tachyarrhythmias in patients with WPW syndrome if carefully managed. PMID- 11424481 TI - [A new bite block for laryngeal mask]. AB - We devised a new bite block made of a used connector of anesthesia machine (ACOMA medical industry CO., LTD.) for laryngeal mask. Fitness for laryngeal mask and strength against patient's biting are the key for its use. Cutting lengthwise the connector (the outside diameter 22 mm, inside diameter 15-19 mm, 55 mm in length) we made a bite block for laryngeal mask. We studied the strength of a new bite block experimentally and recognized its ability to bear the human biting. We conclude a new bite block for laryngeal mask is clinically useful and can be used during anesthesia for its fitness and safety. PMID- 11424482 TI - [New study on the history of anesthesiology (8)--Etymological consideration on a Japanese word "Masui"]. AB - The author briefly describes etymology of a Japanese word "Masui" ([symbol: see text]) and discusses why this word has not been correctly understood by lay people as well as Japanese anesthesiologists. The word "Masui" was coined by Seikei Sugita in 1850 when he translated Dutch edition of Schelesinger's monograph on ether anesthesia into Japanese. Therefore the word is not of Chinese origin and has subsequently been exported to China and the countries of Korean peninsula. Although half a century has elapsed since the first Department of Anesthesiology was established at the Tokyo University Faculty of Medicine, social acceptance for our specialty has not adequately and widely been achieved. The author thinks that one of the causes for this inadequate acceptance is that the correct Japanese words of "Masuika-gaku" and "Masuika-i" for Anesthesiology and Anesthesiologists were not coined and the incorrect words as "Masuigaku" and "Masui-i" have been used. Not a small number of Japanese anesthesiologists still employ the words "Masuigaku" and "Masui-i" without any special reasons. PMID- 11424483 TI - [Tsutsugamushi disease (scrub typhus) in Japan: epidemiological aspects]. AB - Epidemiological aspects of tsutsugamushi disease (scrub typhus) in Japan in 1998 were analyzed using questionnaires. Four hundred and sixteen scrub typhus cases were reported in 24 prefectures in 1998. The annual number of the patients in 1998 was similar to those in the preceding three years. There was no sex difference. The patients at the age of 51 or greater accounted for 72% of the total cases. Patients engaged in farming and forestry accounted for 32% and 14% cases, respectively. Fifty-six, 21 and 19% were reported in Kyusyu, Kanto and Tohoku-Hokuriku districts, respectively, 96% of the total cases being reported in these 3 districts. Most cases were reported from April through June with some from October through December in Tohoku-Hokuriku districts, while most cases were reported from October through December in other districts, including Kyusyu and Kanto districts. Thus, there was a difference in epidemic seasons among the districts. Serotypes of scrub typhus rickettsia were analyzed by serum antibody titers in the Kyusyu district. The novel Kawasaki and Kuroki types were major strains; however, no geographical difference was seen within the Kyusyu district. Interestingly, 24 cases were diagnosed only by the new serotypes not by the classical serotypes (Kato, Karp and Gilliam) in serological tests. This result suggests that further investigations are required to determine the prevalent serotypes in each district and to improve the serological tests. This was the first comprehensive report of epidemiology of scrub typus in entire Japan. Information obtained in the present study provides deep insight into prediction, diagnosis, treatment and prevention of scrub typhus in Japan. PMID- 11424484 TI - [Tsutsugamushi disease (scrub typhus) in Japan: clinical features]. AB - Clinical features of tsutsugamushi disease (scrub typhus) were analyzed, based on 416 cases reported in Japan in 1998. Three major clinical symptoms: eschar, fever and rash were found in 87%, 98% and 92% of the cases, respectively. Elevated levels of CRP, GOT, GPT and LDH were observed in 96%, 85%, 78% and 91%, respectively. These clinical and laboratory findings were observed in the majority of the cases and considered important for diagnosis. Disseminated intravascular coagulation developed in 21 cases, indicating that scrub typhus can be life threatening. Lymphadenopathy was observed in 51% of the cases. Enlarged lymph nodes were limited to the local sites in 75% of these lymphadenopathy cases and most of these sites were adjacent to eschars. Most eschars were scabbed and located in the abdomen and the lower half of the body, especially the feet. This suggests that these parts are frequently exposed to tsutsugamushi mites. Furthermore, the skin is soft in these parts and covered by cloth. These factors may make it possible for mites to keep biting without being noticed for several hours, long enough for rickettsial transmission. Interestingly, eschar and rash were absent in 14% and 8% of the cases, respectively. This result suggests that the cases without the unique symptoms may have been misdiagnosed as common cold or other febrile illnesses. One hundred and fifty-four suspected cases were not scrub typhus cases by the serological tests. The three major clinical symptoms were present in approximately a half of these negative cases, eschar being observed in approximately 70%. This may suggest the presence of new type of scrub typhus can not be diagnosed by the present laboratory tests. Clinical features of scrub typhus in Japan were well revealed, and information obtained in the present study is useful for improving clinical diagnosis. It should, however, be stressed that there were cases that could not be correctly diagnosed only by the clinical symptoms, suggesting that it is important to improve the serological tests. PMID- 11424485 TI - [Epidemiological analysis on many cases of tsutsugamushi disease found in Hiroshima Prefecture, Japan]. AB - The clinical findings of tsutsugamushi disease and the fauna of trombiculid mites in Hiroshima Prefecture were studied in this report. We reviewed 63 cases of tsutsugamushi disease occurring between 1990 and 1999, and most of cases were contracted in the area around the midportion of the Oota River (55 cases; 87.3%). Of these, 30 cases (47.1%) lived in Asakita-ku in Hiroshima City. Eschar was detected in 33/19 (84.6%) cases, and 97.6% (40/41), 88.9% (16/18) patients showed eruption and lymphadenopathy respectively. Biochemical examination revealed liver dysfunction in 38.1% (8/21) patients. Of the 11 cases examined on peripheral blood smear, atypical lymphocytes were detected in 10 cases (90.9%). Fifty-five cases (90.2%) occurred during the restricted season between September and December each year. The predominance of Leptotrombidium scutellare was verified by collection of trombiculid mites along the basin of the Oota River. Serum antibody titration on a patient in Asakita-ku showed reaction to Kawasaki type antigen definitive to L. scutellare. Therefore, we speculate that L. scutellare is a candidate for the vector of Orientia tsutsugamushi in Hiroshima Prefecture. PMID- 11424486 TI - [Strip immunoblot assay (SIA) with recombinant antigens and synthetic peptide for detection of anti HIV-1 and HIV-2 antibodies]. AB - The serodiagnosis of human immunodeficiency virus (HIV) infection has widely been established by the screening test and the confirmatory test. At present, Western blot (WB) assay is mostly used as the confirmatory test. However, this method has the problem in that the sensitivity and the specificity are not enough. A new confirmatory test "CHIRON RIBA HIV-1/HIV-2 SIA" developed by Chiron Corporation uses an immunoblot enzyme immunoassay technique for detection of anti HIV-1 and/or HIV-2 antibodies. This assay employs four recombinant viral antigens (gp120, gp41, p24/p26 and p31) and a synthetic viral antigen (HIV-2 envelope peptide). The characteristic of this method is that the HIV-1 infection and the HIV-2 infection can be differentiated from each other. We therefore compared this SIA method with the WB1 assay for detection of anti HIV-1 antibodies and with the WB2 assay for detection of anti HIV-2 antibodies. Eighty samples from normal adults without HIV infection and known to be negative by three HIV screening tests, respectively, were tested by SIA, WB1 and WB2 assays. The negative rates (specificities) were 97.5%, 80.0% and 87.5% by the SIA, WB1 assay and WB2 assay, respectively. With forty samples from patients without HIV infection but known to be positive by at least one HIV screening test, the negative rates (specificities) were 97.5%, 72.5% and 85.5% by the SIA, WB1 assay and WB2 assay, respectively. The results indicated that the SIA method was more specific than two WB assays. Forty samples from patients with HIV-1 infection and known to be positive by three HIV screening tests, were tested by the SIA and WB1 assay. The positive rates (sensitivities) were 97.5% and 75.0% by the SIA and WB1 assay, respectively. With thirteen samples from patients with HIV-2 infection and known to be positive by three HIV screening test, the positive rates (sensitivities) were 100% and 92.3% by the SIA and WB1 assay, respectively. The results indicated that the SIA method was more sensitive than the WB1 assay. Three sets of sera, which were collected during seroconversion for HIV-1 antibody, were used to compare the positive readings by the SIA and WB1 assay. The SIA method indicated the positive readings earlier than the WB1 assay. The present findings indicated that the SIA method was more specific and sensitive than the WB assay, and would be useful as a confirmatory test. PMID- 11424487 TI - [Functional characterization of a multiple-antibiotic resistant plasmid from clinical isolates of methicillin-resistant Staphylococcus aureus]. AB - During surveillance done as part of the investigation of nosocominal infections due to methicillin-resistant Staphylococcus aureus (MRSA), we have been aware of the close relationship between the presence of certain plasmids and the characteristic patterns of antibiotic resistance. The hypothesis that a mechanism for the rapid and widespread dissemination of resistance to multiple aminoglycosides in clinical isolates of MRSA at our university hospital was the result of transfer of a single plasmid among these strains was examined. About 45% of the total isolates of MRSA (91/200 isolates) carried a 35.5 kb plasmid (designated pCL4) and these isolates always showed resistance to gentamicin, tobramycin, kanamycin, amikacin, astromicin, and arbekacin. Mating experiments between a susceptible strain and resistant MRSA isolates carrying pCL4 showed high frequency transfer of the plasmid. The aminoglycoside resistance patterns of all the transconjugants obtained corresponded well with those of parental strains. However, the plasmid could not necessarily be detected in the transconjugants, whereas the transformants obtained by means of electroporation usually possessed the plasmid. The plasmid-encoded aminoglycoside-resistance determinant, which has been identified as the gene aacA/aphD that encodes the bifunctional enzyme AAC (6')/APH(2"), either in the transconjugants and transformants could be transposed to their chromosomes in the absence of whole plasmid integration resulting from a recombination event. Southern hybridization analysis using an aacA/aphD specific probe demonstrated that there are multiple sites of the insertions indistinguishable among the chromosomes of plasmid-free transconjugants and transformants. These results indicate that rapid dissemination of multiple-aminoglycoside-resistance in nosocominal strains of MRSA resulted from transfer of a conjugative plasmid and has been facilitated by translocations of the resistance gene to the chromosome. PMID- 11424488 TI - [A questionnaire survey on the theory of postoperative infection prophylaxis in gynecology]. AB - A questionnaire survey on the theory of postoperative infection prophylaxis was conducted to obtain the consensus on perioperative antimicrobial use among gynecologists in Japan in the period from April to July 2000. Fifty-six of the 83 gynecologists replied, and the following consensus was obtained. An antimicrobial prophylaxis (AMP) agent should be chosen based on their efficacy against the pathogens expected to be contaminants, such as Staphylococcus spp., Escherichia coli and Bacteroides fragilis group. Use an AMP agent that archives a bactericidal concentrations in both the serum and operating site. Use an AMP agent that has little unfavourable side effects. The newer agents should be considered as a therapeutics for postoperative infections. The therapeutic antimicrobial agents having no cross-resistance to the AMP agents should be used, if postoperative infection is suspected or developed. The most commonly used agent for clean operations are cefazolin (CEZ), followed by cefotiam (CTM) and cefmetazole (CMZ). The most commonly used agent for clean-contaminated operations where low grade level of bacterial invasion expected is CTM, followed by CEZ and CMZ, where as operations where mild grade level of bacterial invasion expected is flomxef (FMOX), followed by CTM and other cephalosporins. PMID- 11424489 TI - [A questionnaire survey on the theory of postoperative infection prophylaxis in orthopedics]. AB - A questionnaire survey on the theory of postoperative infection prophylaxis was conducted to obtain the consensus on perioperative antimicrobial use among orthopedists in Japan in the period from April to September 2000. Fifty of the 91 orthopedists replied, and the following consensus was obtained. An antimicrobial prophylaxis (AMP) agent should be chosen based on their efficacy against the pathogens expected to be contaminants, such as Staphylococcus spp., and Streptococcus spp., Use an AMP agent that achieves a bactericidal concentration in both the serum and operating site. Use an AMP agent that has little unfavourable side effects. Use an AMP agent that affects minimally the normal bacterial flora. The most commonly used agents are the penicillins and first and second generation cephalosporins. The optimal strategy for most commonly used agents entails infusion of the first dose between approximately 30 minutes pre and post-skin incision and the therapeutic levels should be maintained throughout the operation. The AMP agents having no cross-resistance to the prophylactic agents should be used, if postoperative infection is suspected or developed. The most commonly used agent for both clean operations with or without foreign implants and dirty operations is cefazolin (CEZ), followed by cefotiam (CTM) and flomoxef (FMOX). PMID- 11424490 TI - [Serological cross-reaction among Bartonella henselae, Chlamydia pneumoniae and Coxiella burnetii by indirect fluorescence antibody method]. AB - We studied the serological cross-reactions among Bartonella henselae, Chlamydia pneumoniae and Coxiella burnetii by indirect fluorescence antibody (IFA) method, using sera from 8 patients with cat scratch disease (CSD), 13 patients with C. pneumoniae infection and 12 patients with acute Q fever. B. henselae IgG antibody was negative in 13 patients with C. pneumoniae infection, and was positive in 3 (titers being 1:64) of 12 patients with Q fever, whereas B. henselae IgM antibody was negative in all the patients with C. pneumoniae infection or Q fever. C. burnetii IgG antibody was removed by absorption of these 3 sera with C. burnetii antigens, whereas B. henselae IgG antibody did not change. C. pneumoniae IgG antibody was positive in 3 (titers being 1:125 in two, 1:32 in one) of 8 patients with CSD. Both C. pneumoniae and B. henselae IgG antibody titers were significantly reduced by absorption of these 3 sera with B. henselae antigens. C. burnetii IgG or IgM antibodies were negative in all patients with CSD. In conclusion, no serological cross-reaction between B. henselae and C. burnetii was observed. On the other hand. B. henselae IgG antibody cross-reacted to C. pneumoniae antigens, whereas C. pneumoniae IgG antibody did not cross-react to B. henselae antigens. Our findings suggest that determination of B. henselae IgG or IgM antibodies were not influenced by C. pneumoniae and C. burnetii antigens. PMID- 11424491 TI - [Efficacy and safety of influenza vaccination to elderly patients with neurological diseases]. AB - Influenza vaccination is strongly recommended for the elderly persons. Especially elderly patients with neurological diseases are at the high risk because they have more tendencies to develop pneumonia than healthy elderly persons. We vaccinated 105 elderly patients with neurological diseases (cerebrovascular disease, Parkinson disease etc.) and 134 people of a control group. Both groups were inoculated with influenza HA vaccine once. The HI titer increase in elderly patients with neurological diseases was equally good enough in the control group and no significant differences was shown in both groups. No severe side effects and adverse reactions were observed in the elderly patient group. This study shows that influenza vaccination is effective and safe for elderly patients with neurological diseases as the well as healthy elderly person and the HI titer increase after a single influenza vaccine injection is expected to be effective to protect influenza infection. PMID- 11424492 TI - [Early operation for acute type a closing dissection]. AB - From March 1997 to January 2000, we operated eleven cases of Stanford type A acute closing dissection. The patients consisted of 4 men and 7 women with a mean age of 71 +/- 9 years. There were 9 cases (81%) of cardiac tamponade and 5 cases (45%) was in the shock state. There were no malperfusion and end organ ischemia. All cases were operated with deep hypothermia and circulatory arrest. Ascending aortic replacement were performed in 9 cases and 2 cases were performed total arch replacement. 6 cases (63%) were not required blood transfusion. There was one operative death and one hospital death. These result suggest that we had better to perform immediate graft replacement for Stanford type A acute closing dissection as soon as possible, even if there were no serious complications. PMID- 11424493 TI - [Prediction of the need for mechanical ventilation after transsternal thymectomy in patients with myasthenia gravis]. AB - Between June 1992 and May 2000, transsternal extended thymectomy was performed for 70 patients with myasthenia gravis in our hospital. We were able to evaluate 64 of them in terms of prediction of the need for postoperative mechanical ventilation using the score systems reported by Leventhal et al., Kimura et al. and the criteria of Adachi et al.. For these systems, the rates of agreement between predictions and results were 85.9%, 82.8%, and 64.1%, respectively. The two former systems had some false negative cases (i.e., they predicted that ventilation would not be needed when in fact it was), but the last one gave no false negatives. We recommend Adachi's criteria for clinical safety. In our cases the patients whose value of %VC multiplied by FEV1.0% was less than 7,000 (Adachi's criterion is less in 8,300), especially, needed careful management with regard to respiratory crisis. PMID- 11424494 TI - [Early and mid-term results after cardiac or thoracic aortic surgery in patients aged 70 years and older--survival and quality of life]. AB - The early and mid-term survival after cardiac or thoracic aortic surgery and influence of age to the operative mortality were examined in 168 consecutive patients aged 70 years and older from August 1994 to December 1998, together with assessment of postoperative quality of life (QOL). The mean age was 74.1 +/- 3.7 years old (70 to 86 years). 80 patients had IHD, 59 had VHD, 28 had TAA. Preoperative risk score was classified in 5 groups (good, fair, poor, high, extremely high) based on Parsonnet method. Current QOL of the survivors was assessed using Asanoi method with questionnaire by a letter. There were 9 early deaths (5.4%). Following the Parsonnet model observed mortality was absolutely lower than predicted mortality. When age score was excluded in the Parsonnet model, the observed mortality became almost equal with the predicted mortality. There were 22 late deaths (6.9%/P-Y). The actuarial survival rate of age 70 to 74 group was significantly higher than the age 75 years and older group (p = 0.0021). The actuarial survival rate of TAA group was significantly lower than the VHD or IHD group (p < 0.02). Postoperative NYHA and activity score of TAA group were better than VHD or IHD group. We got satisfactory answers for operation in 95% current survivors. Patients aged 70 years and older will be undergone cardiac or thoracic aortic surgery at a reasonable risk and well satisfaction. PMID- 11424495 TI - [Efficacy of modified ultrafiltration in coronary artery bypass grafting]. AB - We evaluated the efficacy of modified ultrafiltration (MUF) in coronary artery bypass grafting. Twenty patients were divide into two groups consisting of a control group (n = 11) and a MUF group (n = 9). MUF was carried out for fifteen minutes immediately after the completion of cardiopulmonary bypass. The blood flow through the ultrafilter was 300 ml/min and about 1,200 ml of water was removed. The hematocrit elevated significantly from 25% to 30% in the MUF group (p < 0.01). Postoperative blood loss in the first 24 hours in the MUF group was significantly less than that in the control group (8 +/- 2 ml/kg vs 12 +/- 4 ml/kg, p < 0.01). There was no statistical difference in the percentage of the increase in body weight after the operation, inflammatory reaction and pulmonary function (A-a DO2, PaO2/FIO2 and duration of intubation) between two groups. In conclusion, MUF is useful to hemoconcentrate and reduce postoperative blood loss in coronary artery bypass grafting. PMID- 11424496 TI - [Extraction of di-2-ethylhexyl phthalate from a cardiopulmonary bypass circuit]. AB - We measured levels of di-2-ethylhexyl phthalate (DEHP) in a priming solution of a cardiopulmonary bypass circuit. The circuit consisted of a venous reservoir, an oxygenator, and polyvinyl chloride (PVC) tubes. Eight circuits were used in this study. In 4 circuits a heparin-coating PVC tube was used and, in the others, a non-coating PVC tube. After they were primed with 1,500 ml of saline, the saline was circulated for 30 minutes at a rate of 6 l/min at 42 degrees C. Another four samples were collected directly from the saline in a polyethylene container as a control. Then the concentrations of DEHP in them were determined by means of gas chromatography. No detectable DEHP was found in controls. But, a small amount was detected in the saline from the circuits (2.75 +/- 4.27 ppb, and 3.75 +/- 4.99 ppb, respectively). DEHP was considered to be leached from the circuit to the priming solution because the tubes were made of PVC containing DEHP as a plasticizer. We suppose the DEHP concentration is probably at a safe level. However, the data on the endocrinologically toxic level of blood DEHP are insufficient. Therefore, the use of plastic in a circuit may require closer scrutiny to determine whether DEHP leached from PVC contributes to exposure to xenoestrogens. PMID- 11424497 TI - [Peri-operative management in patients with thoracic aortic aneurysm complicated with cardiac tamponade]. AB - We retrospectively summarized the peri-operative hemodynamic status and respiratory function in patients with thoracic aneurysms complicated with cardiac tamponade (Tamponade Group; n = 6) in comparison with patients complicated with hemorrhagic pericardial effusion without symptom of cardiac tamponade (Effusion Group; n = 10). There were no difference with regard to preoperative heart rate, pericardial effusion quantity, frequency of emergent operation and operation time. Preoperative hemodynamic status was worse in Tamponade Group than in Effusion Group. The cardiac indices of 12 h, 24 h, 48 h after operation in Tamponade Group were 2.60 +/- 0.60, 2.29 +/- 0.42, 1.95 +/- 0.33 l/min/m2 respectively, those were not different in Effusion Group. The differences between O2 tension in alveolar and arterial blood O2 tension (A-aDO2) and oxygenation indices (OI) of 12 h and 24 h after operation were significantly worse in Tamponade Group than in Effusion Group, but those of 48 h were not different. The operative mortality was 50% in Tamponade Group, and none in Effusion Group. It is very important to maintain hemodynamic status before operation, to perform complete repair (graft replacement) as early as possible and to care intensively after operation for thoracic aortic aneurysm complicated with cardiac tamponade. PMID- 11424498 TI - [Cryopreservation of vascular mixed cell for tissue engineering in cardiovascular surgery]. AB - Tissue engineering (TE) is a new discipline that offers the potential to create replacement structures from autologous cells and biodegradable polymer scaffold. Various vascular and valvular grafts have been tried to create with this TE approach. In clinical use of this technique, harvested and cultured cells have to keep viability until implantation as tissue engineered tissue. But few research for cryopreservation of vascular mixed cells has been performed. So, we investigated the proper method for cryopreservation of vascular mixed cells harvested from femoral artery and vein of dogs. Cells were cultured and divide into three groups, A: cryopreserving in 5% dimethylsulfoxide (DMSO), hydroxyethyl starch (HES), and fetal bovine serum (FBS) with -80 degrees C freezer; B: cryopreserving in 10% DMSO and FBS with programmed freezer; C: control (continuous culture in media). After rapid thawing at 40 degrees C, group A showed higher viability than group B with flow cytometry. The results means that vascular mixed cells can be successfully cryopreserved in the DMSO/HES mixture simply and inexpensively, without rate controlled freezing. PMID- 11424499 TI - [A new instrument for insertion of anvil head for circular stapled anastomosis]. AB - Difficulty associated with the insertion of the anvil head into the esophageal stump, caused by narrowing of the cervical esophagus, is a big problem for staple anastomosis during esophageal reconstruction. Postoperative cervical stricture also has been a major complication of esophagogastric anastomosis using a circular stapler. The stricture results from esophageal laceration caused by the anvil insertion. To avoid these complications, we designed a new retractor with triple-flap arms, which spreads out in a concentric circle from 20 mm to 35 mm in diameter. This instrument is useful for gently expanding the cervical esophagus without lacerations, and facilitates an anvil insertion into the stump of esophagus. In the 8 patients in whom this instrument was applied for dilation, none of the patients suffered from any stricture. Our clinical result indicates the usefulness of this new instrument for avoiding esophageal laceration and preventing stricture of esophageal anastomosis performed by circular stapler. PMID- 11424500 TI - [Blue toe syndrome after coronary artery bypass grafting]. AB - We report three cases of blue toe syndrome (BTS) after coronary artery bypass grafting (CABG). All patients were cyanotic and exhibited painful toes two to four weeks after CABG. They were treated with antiplatelet and anticoagulant agents, and one patient underwent replacement of the abdominal aorta. Thus, BTS may occur after CABG with coronary angiography, extra-corporeal circulation or intraaortic balloon pumping. For the treatment of BTS, surgery remains the most effective option. PMID- 11424501 TI - [A case of benign schwannoma arising in the brachial plexus with intrathoracic extension]. AB - A case of benign schwannoma originating from the lowest trunk of the left brachial plexus with intrathoracic extension was reported. Intrathoracic growth of a schwannoma of the brachial plexus has been reported in only five cases in the literature. The patient was a 35-year-old man and had been pointed out an abnormal shadow at the left lung apex on the roentgenogram since 20-year-old. The tumor shadow was increased to seven cm from four in size on recent chest X-ray. We performed a video-assisted thoracoscopic surgery with the oblique skin incision at the left neck and removed the tumor completely and safely. PMID- 11424502 TI - [Surgical treatment for pulmonary arterial injury caused by sewing needles]. AB - A 41-year-old woman was found lying in her home, which was on fire, and was brought to our hospital by ambulance. Her complaint was severe dyspnea, and a respirator was needed. Contusion and abrasion were present on her anterior chest. A roentgenogram of her chest disclosed cardiac enlargement and two linear metallic shadows. A computed tomographic scan of the chest showed cardiac tamponade and a needle from the chest wall to the main pulmonary artery. Through a median sternotomy approach, the pericardium was opened. About 200 ml of blood was pooled in the pericardium cavity, and a needle was found in the main pulmonary artery. The needle was removed and the bleeding point was closed using suturing with 5-0 prolene without cardiopulmonary bypass. PMID- 11424503 TI - [A case of thrombosed St. Jude Medical valve 16 years after initial mitral valve replacement]. AB - We report successful surgery for a thrombosed St. Jude Medical (SJM) valve 16 years after the initial mitral valve replacement even under conditions of satisfactory anticoagulation therapy. A 61-year-old-female had intermittent claudication and was admitted to our hospital for examination. The prosthetic valve sounds were normal to auscultation and the left ankle-pressure index was decreased to 0.6. Transthoracic echocardiography revealed no mitral regurgitation and a mean mitral valve gradient of 6-7 mmHg. Furthermore, transesophageal echocardiography revealed that one of the leaflets of the prosthetic valve was entirely immobilized at the closing position and a mobile soft tissue mass, 5 mm in diameter, was detected at the atrial side of the obstructed leaflet. Although 96,0000 IU of urokinase was administered intravenously for a week, we could not confirm any change in leaflet mobility. At the time of surgery, the posterior leaflet of the SJM valve, which was implanted at an anatomical orientation, was obstructed at the closing position with old and fresh thrombi. We decided upon replacement with a CarboMedics 29 M prosthetic valve. Postoperative medication consisted of warfarin plus low-dose aspirin. Generally, valve thrombosis occurs within 5 years after valve replacement. However, valve thrombosis is possible even in a reliable SJM valve and as long as 16 years after replacement. Therefore, the implantation of an SJM valve at an anti-anatomical orientation might lower the incidence of valve thrombosis in addition to life-long anticoagulation therapy. PMID- 11424504 TI - [A surgical treatment for ischemic heart disease associated with systemic lupus erythematodes: report of a case]. AB - We reported a case of coronary artery bypass grafting (CABG) associated with systemic lupus erythematodes (SLE). A 45-year-old male who had been treated for SLE with prednisolone for 9 years was transferred to our department for a surgical treatment due to ischemic heart disease (IHD). We successfully performed CABG resolving various perioperative complications. The intensive care is indispensable in the case of IHD with SLE. PMID- 11424505 TI - [A case report of primary cardiac leiomyosarcoma]. AB - A 26-year-old woman presenting symptoms of rapid progressive heart failure consulted with our hospital. Two-dimensional echocardiography showed a large mass in the left atrium (LA) and severe pulmonary hypertension. Emergent operation was performed because of hemodynamic reason and the risk of embolism. The tumor (7 x 6 x 3 cm) which was lobulated and had a smooth surface was arising from posterior wall of LA extending into the ostium of right lower pulmonary vein. The portion of tumor located was excised. The defect was reconstructed by bovine pericardium. Postoperative histopathological diagnosis was leiomyosarcoma. Hemodynamics recovered after surgical treatment and there were no problems for 5 months. However, tumor recurrence in LA and pulmonary metastasis was found 5.5 months after surgery. The patient died two weeks later by progressive heart failure. Primary cardiac leiomyosarcoma is extremely rare. The current world literature is reviewed with respect to this rare tumor. PMID- 11424506 TI - [A case of collision tumor involving the mouth of the left lower lobe bronchus treated by left lower sleeve lobectomy]. AB - A 72-year-old man was admitted because of hemoptysis. Further examination revealed adenocarcinoma involving the orifice of the left lower lobe bronchus. We performed left sleeve lower lobectomy and the postoperative pathologic examination revealed a collision tumor comprising adenocarcinoma and squamous cell carcinoma. PMID- 11424507 TI - [Surgically treated cases of pulmonary arteriovenous fistula]. AB - We report three cases of pulmonary arteriovenous fistula (PAVF) in which we performed limited resection. Case 1: female, 75 of age. CT and pulmonary angiography revealed PAVF of 2 cm in diameter in the right S8, and partial resection was performed. Case 2: female, 45 of age. CT and pulmonary angiography revealed PAVF of 7 cm in diameter in the right S10, and occlusion of fistula was performed. Case 3: male, 43 of age. CT pulmonary angiography and 3 D-CT revealed PAVF of 3 cm in diameter in the right S6, and occlusion of fistula was performed. The last case was accompanied by Rendu-Osler-Weber disease. Three patients showed a good postoperative course, without developing new fistula. Occlusion of fistula allows to close feeding artery and draining vein safely and securely. Considering invasion and preservation of pulmonary function after surgery, occlusion of fistula may be a primary option for treating PAVF. PMID- 11424508 TI - [Mediastinal lymph node carcinoma without apparent primary lesion: report of case]. AB - A 62-year-old man with dyspnea had an enlarged lymph node, 6.5 cm in size in the left hilum, but no primary carcinoma could be found. The patient underwent lymph node dissection under video-assisted thoracoscopic surgery, and the histological diagnosis was undifferentiated carcinoma. No primary lesion has been identified even in postoperative survey. The postoperative course was uneventful with no evidence of recurrence eight months after the operation. PMID- 11424509 TI - Rights-based food and nutrition perspective: 21st century challenge for dietetics. AB - Progress in addressing the problems of hunger and malnutrition has been slow and uneven across regions and countries of the world. A rights-based perspective, not merely a utilitarian argument, must serve as the rationale for humans to be properly nourished. Rights impose corresponding binding obligations on others and the concept of rights holders and duty bearers goes beyond the human development paradigm. The right to adequate food is realized when every man, woman and child, alone or in community with others, has physical and economic access at all times to adequate food or means for its procurement. As duty bearers, governments are obligated to assure the realization of that right and accountable if the right is violated. The use of the human rights approach to adequate food necessarily implies a re-examination and fundamental changes in the way we think about hunger and malnutrition and their causes; the manner of shaping and reshaping policies; the context and processes of planning, monitoring and evaluating programmes; and the substance and method of research and education in food and nutrition. Dietitians and dietetic associations can be instrumental in making the human right to adequate food a reality. PMID- 11424510 TI - The effect of weekly iron supplementation on anaemia and on iron deficiency among female tea pluckers in Bangladesh. AB - AIM: To investigate the effect of weekly iron supplementation on anaemia and iron deficiency among adult, female tea pluckers. METHOD: A randomized double-blind intervention trial was conducted in a tea estate in Bangladesh where a total of 280 women received either weekly iron supplementation (200 mg ferrous fumarate and 200 mg folic acid) for 24 weeks or a matching placebo. Capillary blood samples were drawn at baseline and post-trial to determine haemoglobin, haematocrit and ferritin concentration. Mean corpuscular haemoglobin concentration (MCHC) was calculated using the haemoglobin and haematocrit values. RESULTS: The mean haemoglobin concentration in the supplemented group increased by 5.52 g L-1 over the study period, on average, while ferritin values decreased by 0.33 microgram L-1. The control group showed a decrease in both mean haemoglobin (-0.24 g L-1) and ferritin (-5.32 micrograms L-1). Those individuals in the supplemented group with the lowest pretrial haemoglobin and ferritin values experienced the greatest improvements post-trial, whereas nonanaemic individuals showed a decrease in both haemoglobin and ferritin concentrations. A total of 62.2% of women in the supplemented group reported feeling better and more energetic compared to 51.1% in the placebo group; 14.4% of the supplemented group and 22.7% of the control group complained about side-effects. CONCLUSION: Weekly iron supplementation was logistically simpler and cheaper than daily supplementation but would have to be continued on a longer term basis in order to combat both anaemia and iron deficiency. PMID- 11424511 TI - Use of 'light' foods and drinks in French adults: biological, anthropometric and nutritional correlates. AB - BACKGROUND: A population of over 12,000 mature subjects participated in a longitudinal study (8 years) of nutrition and health (the Su.Vi.Max Study). In this context, a specific cross-sectional study was carried out in a randomly selected subpopulation. AIM: To identify anthropometric, nutritional and biochemical correlates of spontaneous use of 'light' foods and drinks in a free living population. DESIGN: Men (n = 2299) and women (n = 1979), 45-60 years, reported their food intakes over six non-consecutive days. Consumers of low-fat and low-sugar foods and drinks, and artificial sweeteners, were compared with non consumers. RESULTS: Users of low-sugar products were heavier than non-users; female consumers of low-fat products, but not males, had higher body weight and BMI than non-consumers. Users of low-sugar products had higher triacylglycerols and glycaemia than non-users while biochemical parameters were not different in users and non-users of low-fat products. Use of low-sugar products led to increased diet density of a few micronutrients, including cholesterol. Low-fat product selection was associated with increased intake of most micronutrients, both in absolute value and in density. CONCLUSIONS: In mature adults, selection of fat-reduced products was associated with improved quality of the diet, while anthropometric and biological parameters appeared less favourable in consumers of low-sugar products vs. non-consumers. The longitudinal follow-up of the cohort in future years will help determine cause-and-effect relationships among these parameters. PMID- 11424512 TI - Dietary trends among Scottish schoolchildren in the 1990s. AB - BACKGROUND: In recent years, healthy eating messages have sought to highlight the advantages of a healthy balanced diet, but there is little evidence that the recommendations have been translated into sustained behavioural change. In Scotland, the national diet has become a major focus in key policy documents, and the diet of children and young people has been consistently highlighted as an area of particular concern. This paper reports on dietary trends among Scottish schoolchildren during the 1990s. METHODS: Data were collected from a representative sample of Scottish schoolchildren as part of the WHO Cross national Health Behaviour of School-aged Children (HBSC) study. Three consecutive national surveys were undertaken in Scotland, in 1990, 1994 and 1998, using self completion food frequency questionnaires with 11-, 13- and 15-year-old school pupils. RESULTS: Between 1990 and 1998, fruit and vegetable consumption increased among Scottish schoolchildren, especially among girls, but levels of consumption fell below current dietary recommendations. There has been a concomitant increase in consumption of high-fat and high-sugar foods, and consumption of these foods is higher among boys and children from lower socio-economic groups. CONCLUSION: The findings emphasize the need for continued health promotion efforts to improve the diet of schoolchildren in Scotland especially among lower socio-economic groups. PMID- 11424513 TI - 'Change to cup': an audit to determine parental awareness and practices in changing from bottle to cup. AB - BACKGROUND: It is recommended that infants from the age of 6 months should be introduced to drinking from a cup and the use of a bottle should be actively discouraged after the age of 1 year. The number of young children using a feeder bottle after the age of 1 year is not known but it is recognized by health professionals as a 'growing' concern. AIMS: To obtain local information about the number of children using a bottle past the age of 1 year, what fluids were being offered and to assess parental awareness about when a cup can be introduced. METHODS: The study data was collected by questionnaire from 407 parents between June 1998 and January 1999. Socially disadvantaged groups were targeted for the sample group as well as care being taken to include a representative ethnic population. RESULTS: Ninety-nine per cent of the study infants were below the age of 2 years, with the age range being 12-28 months. Thirty per cent were using only a cup/beaker, 8% of infants still used a bottle only, and 62% used both a bottle and a cup. Drinks/food other than milk and water had been offered from the bottle by 47% of respondents. More than 60% of the parents thought that the age at which the use of a bottle should be discontinued was older than that recommended. Only 18% thought a cup/beaker could be introduced as early as 6 months. CONCLUSION: The results show that there is a need to actively encourage the use of a cup at an earlier age and to discourage the inappropriate and extended use of a feeder bottle. Barriers to change need to be addressed. PMID- 11424514 TI - What nutritional support literature do hospital nursing staff require? AB - BACKGROUND: Many patients require nutritional intervention to help treat and/or prevent malnutrition. Nursing staff play a key role in identifying malnourished patients. To improve the management of artificial nutritional support, the Hammersmith Hospital NHS Trust has produced guidelines. This audit aimed to assess the need and adequacy of these guidelines and to examine any additional information required. METHODS: The dietetic department and nursing practice group developed a questionnaire. A total of 260 questionnaires were distributed by dietitians, 10 copies of the questionnaire to nurses on 26 wards. The completed forms were returned by internal post or collected the dietitian. RESULTS: The response rate was 29%. All 77 nurses who responded felt there was a need for nutrition literature. Seventy-three per cent of nurses were aware that the guidelines existed; however, referral to the resource was poor. Requests for additional information encompassed tube feeding and catering information. To provide this information, 84% requested a nutrition tutorial, 81% a preprinted care plan and 96% a ward manual specific to their speciality. CONCLUSIONS: All nurses who were surveyed requested nutrition information. The current guidelines already reflect the requested information, but usage is poor. We aim to improve access and awareness by the hospital intranet, nutrition education and induction programme. The next update will cover gaps identified and additional requirements outside the guidelines remit will be discussed with the nursing practice group. PMID- 11424515 TI - Dietary treatment of irritable bowel syndrome: current evidence and guidelines for future practice. AB - The aim of this literature review is to produce guidelines for dietetic practice in irritable bowel syndrome (IBS) by evaluating the research available. In this area randomized control trials (RCT) only account for a small proportion of the literature and have been concentrated in the modification of dietary fibre in patients with IBS. The bulk of the literature is mainly observational trials from which no indisputable conclusions can be extracted. In this review, the evidence available has been interpreted within the context of the current knowledge base. Conclusions are drawn to facilitate the development of guidelines, enabling a starting point for discussion and an evaluation of current practice. The literature available on therapeutic dietary manipulation in IBS patients is centred around non-starch polysaccharides (NSPs), mono and disaccharide sensitivity and food intolerance. The production of these guidelines has focused on research examining the role of dietary components in the therapeutic management of patients with IBS. However, where there is a deficiency in the literature directly relating dietary intake to management of IBS patients, physiological function in relation to dietary components has been relied upon to produce practical guidelines which can be applied realistically in a clinical environment. An interpretation of the evidence has revealed a limited role for exclusion diets, a move away from high-fibre diets towards the manipulation of fibre fractions in the diet, an evaluation of the effects of caffeine on gut function and the necessity for individual dietary assessment to identify dietary issues pertinent to the patient's symptoms. These guidelines outline a positive role for dietitians in the treatment of IBS patients which draws on the unique skills possessed by dietitians regarding the assessment of habitual eating habits and therapeutic dietary manipulation. PMID- 11424516 TI - Weight loss and delayed gastric emptying following a South American herbal preparation in overweight patients. AB - BACKGROUND: Obesity and overweight may soon affect more than half of the population in some regions of the world and are associated with diabetes, hypertension and other diseases that cause morbidity, mortality and high health care expenditure. No one approach, whether dietetic management, medication, or commercial weight loss programme, can alone solve the problem--all potential treatments need to be investigated and exploited. Among the herbal preparations known to non-western cultures are materials which may have applications in modulating physiological processes which influence gut motility, food intake and energy balance. One such mixed herbal preparation is 'YGD' containing Yerbe Mate (leaves of Ilex paraguayenis), Guarana (seeds of Paullinia cupana) and Damiana (leaves of Turnera diffusa var. aphrodisiaca). AIMS: This study had two distinct aims: to determine the effect of a herbal preparation 'YGD' containing Yerbe Mate, Guarana and Damiana on gastric emptying; to determine the effect of the same preparation on weight loss over 10 days and 45 days and weight maintenance over 12 months. METHODS: Gastric emptying was observed using ultrasound scanning in seven healthy volunteers following YGD and placebo capsules taken with 420 mL apple juice. Body weight was observed before and after 10 days of treatment with three YGD capsules or three placebo capsules before each meal for 10 days in 44 healthy overweight patients attending a primary health care centre. Forty-seven healthy overweight patients entered a double-blind placebo-controlled parallel trial of three capsules of YGD capsules before each main meal for 45 days compared with three placebo capsules on body weight. Body weight was monitored in 22 patients who continued active (YGD capsules) treatment for 12 months. RESULTS: The herb preparation YGD was followed by a prolonged gastric emptying time of 58 +/- 15 min compared to 38 +/- 7.6 min after placebo (P = 0.025). Body weight reductions were 0.8 +/- 0.05 kg after YGD capsules compared to 0.3 +/- 0.03 kg after placebo capsules over 10 days, and 5.1 +/- 0.5 kg after PGD capsules compared to 0.3 +/- 0.08 kg after placebo over 45 days. Active treatment with YGD capsules resulted in weight maintenance of the group (73 kg at the beginning and 72.5 kg at the end of 12 months). CONCLUSIONS: The herbal preparation, YGD capsules, significantly delayed gastric emptying, reduced the time to perceived gastric fullness and induced significant weight loss over 45 days in overweight patients treated in a primary health care context. Maintenance treatment given in an uncontrolled context resulted in no further weight loss, nor weight regain in the group as a whole. The herbal preparation is thus shown to be one that significantly modulates gastric emptying. Further clinical studies with dietetic monitoring of energy intake, dietary quality, satiety ratings, body weight and body composition are now indicated, and examination of the active principles contained in the three herbal components may prove rewarding. PMID- 11424517 TI - Attitudes towards infant feeding among adults in a low socioeconomic community: what social support is there for breastfeeding? AB - An analysis of the role of social support in influencing breastfeeding in a low socioeconomic area in South Australia was undertaken by examining infant feeding attitudes and experiences of mothers, fathers and grandmothers as well as the general community. A random telephone survey of over 3,400 adults (including a more extensive survey of 373 mothers, fathers and grandmothers in the sample) in this area indicated that there was little support for breastfeeding compared to bottle-feeding with similar barriers to breastfeeding found in all target groups as well as the general community. These included breastfeeding in public, the convenience of bottle-feeding, maternal discomfort of breastfeeding, the support required for breastfeeding, fathers' involvement with feeding, and a mother's previous experience of breastfeeding. Strategies promoting and supporting breastfeeding should address these issues and should be directed at the community in general rather than specific groups within the community. PMID- 11424518 TI - What is normal? A study of normal breastfeeding dyads during the first sixty hours of life. AB - The aim of the study was to describe breastfeeding behaviour during the first 60 hours of life of 'normal' dyads. The Infant Breastfeeding Assessment Tool (IBFAT) was used on a convenience sample of 37 mother-baby dyads. Individual dyads were assessed and then the group was combined to seek underlying patterns. Average time between feeds was 3.36 +/- 0.17 hours. There was a marked diurnal pattern of feeding. Parity affected the rate at which high feeding ability scores were achieved, while the majority of babies were achieving high scores within 24 hours of birth. The results provide a baseline against which future research on interventions during labour and its effect on breastfeeding initiation can be compared. PMID- 11424519 TI - Mastitis in lactating women: physiology or pathology? AB - Mastitis is a significant problem amongst lactating women yet there remains a paucity of scientific research into the anatomical, physiological and pathological determinants for mastitis. There is also scant knowledge regarding the physiological changes occurring within the breast as a result of mastitis. This paper examines the available research and current clinical and scientific opinion concerning the breast's response to inflammation and infection and the numerous influences that may impact upon the development of mastitis. In particular, the difficulties associated with differentiating between infective and non-infective mastitis are discussed. PMID- 11424520 TI - [Pyridine carboxylic acids in the intracellular regulation of reproduction in ciliates]. PMID- 11424521 TI - [Proteinase inhibitors in the anterior midgut of cockroach Nauphoeta cinerea]. PMID- 11424522 TI - [Cholinesterase active sites. Statistical analysis of the structure variability]. PMID- 11424523 TI - [Lactate and malate dehydrogenase activity in the sterlet and Russian sturgeon under conditions of the widespread muscular pathology]. PMID- 11424524 TI - [Purine biosynthesis de novo in the retina: evolutionary aspects]. PMID- 11424525 TI - [Development of the hydrocarbon digestive-transport 'assembly line' during ontogenesis in rats]. PMID- 11424526 TI - [Interrelations between temporal parameters of the calling signal in the cricket Gryllus bimaculatus]. PMID- 11424527 TI - [Oscillations of the membrane potential induced by excitatory amino acids in isolated spinal neurons in Lampetra fluviatilis]. PMID- 11424528 TI - [Regional and species heterogeneity of the vascular response: cholinergic muscarinic regulation of the motor function of blood vessels in rats and frog]. PMID- 11424529 TI - [Sex dimorphism in saccharin consumption during development in rats. Age dependent effect of maternal deprivation]. PMID- 11424530 TI - [Localization of glutamate in the nervous system of Drosophila melanogaster: immunocytochemical study]. PMID- 11424531 TI - [Chloride cells--a component of the fish olfactory epithelium]. PMID- 11424532 TI - [Effect of inositol triphosphate receptor blocker heparin on contractions in the Lampetra fluviatilis muscle]. PMID- 11424533 TI - Outsourcing health care information technology. PMID- 11424534 TI - Outsourcing must be a global strategy--not a technology fix. PMID- 11424535 TI - "Top 10" future trends in outsourcing. PMID- 11424536 TI - A super-sized problem: restaurant chains piling on the food. PMID- 11424537 TI - Eating disorders: all in the family? PMID- 11424538 TI - Tips for contract negotiations and establishing MNT rates. PMID- 11424539 TI - Regulation of nutrition professionals continued by states. PMID- 11424540 TI - Ensuring healthy and well-nourished children. PMID- 11424541 TI - The fifth edition of the Dietary Guidelines for Americans: lessons learned along the way. PMID- 11424542 TI - Reliability and validity of the Child and Adolescent Trial for Cardiovascular Health (CATCH) Food Checklist: a self-report instrument to measure fat and sodium intake by middle school students. AB - OBJECTIVE: To develop a scoring algorithm and evaluate the reliability and validity of scores from the Child and Adolescent Trial for Cardiovascular Health (CATCH) Food Checklist (CFC) as measures of total fat, saturated fat, and sodium intake in middle school students. DESIGN: Randomized, controlled trial in which participants were assigned to 1 of 3 study protocols that varied the order of CFC and 24-hour dietary recall administration. Criterion outcomes were percent energy from total fat, percent energy from saturated fat, and sodium intake in milligrams. SUBJECTS/SETTING: A multiethnic sample (33% ethnic and racial minorities) of 365 seventh-grade students from 8 schools in 4 states. STATISTICAL ANALYSES: Multivariable regression models were used to calibrate the effects of individual food checklist items; bootstrap estimates were used for cross validation; and kappa statistics, Pearson correlations, t tests, and effect sizes were employed to assess reliability and validity. RESULTS: The median same-day test-retest reliability kappa for the 40 individual CFC food items was 0.85. With respect to item validity, the median kappa statistic comparing student choices to those identified by staff dietitians was 0.54. Test-retest reliability coefficients ranged from 0.84 to 0.89 for CFC total nutrient scores. Correlations between CFC scores and 24-hour recall values were 0.36 for total fat, 0.36 for saturated fat, and 0.34 for sodium; CFC scores were consistent with hypothesized gender differences in nutrient intake. APPLICATIONS/CONCLUSIONS: The CFC is a reliable and valid tool for measuring fat, saturated fat, and sodium intake in middle school students. Its brevity and ease of administration make the CFC a cost-effective way to measure middle school students' previous day's intake of selected nutrients in school surveys and intervention studies. PMID- 11424543 TI - Screening method evaluated by nutritional status measurements can be used to detect malnourishment in chronic obstructive pulmonary disease. AB - OBJECTIVE: To evaluate and develop a screening method for malnutrition among patients with chronic obstructive pulmonary disease (COPD). DESIGN: Findings from a screening sheet for malnutrition were compared with results from full nutritional assessment. The screening sheet included 7 questions regarding body mass index, anorexia, loss of weight, and other variables possibly affecting nutritional status. Each answer was assigned a point value, and a total of 4 and 5 points were tested as criterion for malnutrition. Full nutrition assessment included measurements of weight and height (body mass index), serum albumin and prealbumin, total lymphocyte count, triceps skinfold thickness, mid-arm muscle circumference or area, and information on unintentional weight loss. Malnutrition was defined by 3 or more values below reference values. SUBJECTS: Randomly selected patients (n = 34) with a clinical diagnosis of COPD, 15% of eligible patients admitted to the Department of Lung Medicine at National University Hospital, Reykjavik, Iceland, during the time of the study. STATISTICAL ANALYSIS: Sensitivity, specificity, and predictive values were calculated to evaluate the screening sheet. Each of the 7 parameters used in a full nutrition assessment was similarly evaluated as an indicator to predict malnutrition. RESULTS: Full nutrition assessment identified 13 of 34 patients (38%) as malnourished. Using 4 points as a criterion for malnutrition, the screening sheet to be used for patients with COPD resulted in sensitivity of 0.69 and specificity of 0.90. CONCLUSION: The results confirm the frequent finding of malnutrition among patients with COPD and show that a simple screening sheet can be used to identify which patients need further nutrition assessment and treatment. PMID- 11424544 TI - Ginseng does not enhance psychological well-being in healthy, young adults: results of a double-blind, placebo-controlled, randomized clinical trial. AB - OBJECTIVE: Ginseng is a popular, commercially available dietary supplement that is purported to have a number of psychological benefits. The purpose of this study was to examine these claims, with specific reference to ginseng's effects on affect and mood. DESIGN: Prospective, double-blind, placebo-controlled, randomized clinical trial. PARTICIPANTS/SETTING: Eighty-three adults (40 women, 43 men) participated in this study (mean age = 25.7 year). Participants were recruited from within a university community and at area health clubs. INTERVENTION: Participants were randomly assigned to one of three experimental conditions: placebo (lactose), 200 mg ginseng, or 400 mg ginseng. The ginseng preparation used in this study consisted of the Panax ginseng C A Meyer concentrate G115 in capsular format. Each participant was given a 60-day allotment of their respective supplement along with written instructions about the proper intake and storage of the capsules during the 8-week study period. MAIN OUTCOME MEASURES: Positive affect, negative affect, and total mood disturbance. Measures were obtained pre- and post-intervention. STATISTICAL ANALYSES PERFORMED: Repeated measures multivariate analysis of variance was used. Because there were three dependent variables, and in an effort to minimize the experimentwise-error rate, alpha was adjusted using the Bonferroni technique (i.e., P < .05/3 = P < .016). RESULTS: Ginseng supplementation had no effect on positive affect, negative affect, or total mood disturbance (all P > .016). CONCLUSION: The present findings do not support claims that chronic ginseng supplementation--at either its clinically recommended level or at twice that level--enhances affect or mood in healthy young adults. PMID- 11424545 TI - Considerations in planning vegan diets: children. AB - This article reviews research on the growth and nutrient intake of vegan children and provides guidelines for counselling parents of vegan children. Although diets of vegan children meet or exceed recommendations for most nutrients, and vegan children have higher intakes of fiber and lower intakes of total fat, saturated fat, and cholesterol than omnivore children, some studies indicate that they may be low in calcium. In addition, bioavailability of zinc and iron from plant foods can be low. Protein needs are slightly higher for vegan children but are easily met with a varied diet that provides adequate energy. Special attention should be given to dietary practices that enhance absorption of zinc and iron from plant foods. Further, good sources of the omega-3 fatty acid linolenic acid should be emphasized to enhance synthesis of the long-chain fatty acid docosahexanoic acid. Dietetics professionals who counsel vegan families should help parents identify good sources of vitamin B-12, riboflavin, zinc, calcium and, if sun exposure is not adequate, vitamin D. This should not be problematic, due to the growing number and availability of fortified vegan foods that can help children meet all nutrient needs. Therefore, with appropriate food choices, vegan diets can be adequate for children at all ages. PMID- 11424546 TI - Considerations in planning vegan diets: infants. AB - Appropriately planned vegan diets can satisfy nutrient needs of infants. The American Dietetic Association and The American Academy of Pediatrics state that vegan diets can promote normal infant growth. It is important for parents to provide appropriate foods for vegan infants, using guidelines like those in this article. Key considerations when working with vegan families include composition of breast milk from vegan women, appropriate breast milk substitutes, supplements, type and amount of dietary fat, and solid food introduction. Growth of vegan infants appears adequate with post-weaning growth related to dietary adequacy. Breast milk composition is similar to that of non-vegetarians except for fat composition. For the first 4 to 6 months, breast milk should be the sole food with soy-based infant formula as an alternative. Commercial soymilk should not be the primary beverage until after age 1 year. Breastfed vegan infants may need supplements of vitamin B-12 if maternal diet is inadequate; older infants may need zinc supplements and reliable sources of iron and vitamins D and B-12. Timing of solid food introduction is similar to that recommended for non vegetarians. Tofu, dried beans, and meat analogs are introduced as protein sources around 7-8 months. Vegan diets can be planned to be nutritionally adequate and support growth for infants. PMID- 11424547 TI - Developing actionable dietary guidance messages: dietary fat as a case study. AB - Although consumers say they are concerned about nutrition and are aware that eating a healthful diet is important for good health, this knowledge does not always translate into healthful diet behaviors or motivate behavior change. In an effort to better understand consumer attitudes about nutrition and to explore alternatives for communicating dietary advice in language that is meaningful and motivates behavior change, the International Food Information Council (IFIC) conducted qualitative research with consumers (using focus groups) and registered dietitians (using telephone interviews) in 1998 and 1999. Results of the research are presented using dietary fat as a case study. Findings from the IFIC research were reported to the Dietary Guidelines Advisory Committee to assist the Committee in developing meaningful and action-oriented dietary advice related to dietary fat for inclusion in the 2000 Dietary Guidelines for Americans that would be motivating and easy for consumers to implement. The recommendation to moderate fat intake in the new dietary guideline, "Choose a diet that is low in saturated fat and cholesterol and moderate in total fat" is consistent with communication recommendations in the IFIC research. Further, the moderate fat message is empowering because it suggests an achievable dietary regimen and reduces guilt and worry about foods. It allows flexibility to enjoy desired foods and promotes using common sense when it comes to diet. Several issues emerged from the IFIC research that apply to general nutrition communications with consumers, whether it be through national nutrition recommendations or in one-on-one counseling situations: to be effective, messages to consumers about nutrition, and specifically dietary fat, must address sources of discomfort about dietary choices; they must engender a sense of empowerment; and they should motivate both by providing clear information that propels toward taking action and appeals to the need to make personal choices. PMID- 11424548 TI - Focus groups indicate that vegetable and fruit consumption by food stamp-eligible Hispanics is affected by children and unfamiliarity with non-traditional foods. PMID- 11424549 TI - The effects of antiretroviral protease inhibitors on serum lipid levels in HIV infected patients. PMID- 11424550 TI - Nutrient intake of infants hospitalized with lower respiratory tract infections. PMID- 11424551 TI - Case problem: dietary treatment for children with hyperlipidemias. PMID- 11424552 TI - Learning needs and continuing professional education activities of Professional Development Portfolio participants. PMID- 11424553 TI - [The examination of the analgetic action of polarized light]. AB - The influence of lowintensive polarized light on pain and nonepain behavioural responses (BR) was studied in experiments on mice with the locus of tonic pain (phormalin test). It was shown, that the action of polarized light on the acupuncture points (AP) statistically reliable decrease of pain BR (lisking of the locus of a pain), but increase the duration of nonepain BR (sleeping, eating). The analgetic effect depends from a choice of AP and ehpozition. The maximal analgesia (50%) was observed after 10-minute action of polarized light on AP E-36. PMID- 11424554 TI - [The role of arachidonic and eicosapentaenoic acid lipoxygenase products in the pathogenesis of generalized parodontosis]. AB - Mix saliva arachidonic acid lypoxygenase products (5-HETE, 15-HETE, LTB4) amount was measured during generalised parodontities treatment by means of a radioimmunological research technique. It is placed, that their mix saliva contents augmentation at parodontities coincides with the deterioration of disease clinical features. Application of Tekom keeping in a great amount eicosapentaenoic acid, leads to mix saliva 5-HETE, 15-HETE, [symbol: see text]TB4 content lowering and parodontium clinical state improvement. PMID- 11424555 TI - [Physiopathological mechanisms of reduced skin capillary resistance in disorders of microcirculation]. AB - Nesteroffs method is a very simple and effective for diagnosis skin capillary resistance, this method is not only effective for diagnosis, but it is very applicable for curing effectiveness control. Women's skim blood vessel conditions were investigated. Only the women of before climacteric period were invavend, all of them had microcirculation which reduced elastins of skin. Reduction of capillary resistance is closely connected with the degree of microcirculation distonia and connected with it skin elastans reduction. We can take into consideration that the changings found are connected with the changings of blood vessel microcirculation. Microcirculation distomia causes the changings of skin elastins. PMID- 11424556 TI - [Methods of visualization of biologically active points]. AB - The results of acupunctural investigation (biologically active points) (BAPs) by using the method of their visualization are presented in this paper. As a result of these investigation it is established that BAPs have concise spatial, disposition which is characteristic to the only one separately taken person and don't change its position in the course of time. Investigation of BAP changes activity in time allowed to determine similar behavior disposition of some points that testify, presumably, that these BAP refer to the same meridian or organ. PMID- 11424557 TI - [Transmembrane ionic currents in smooth muscle cells of rat tail artery]. AB - Whole-cell currents in single smooth muscle cells freshly isolated from the rat tail artery have been studied using patch-clamp recordings techniques. Outward current evoked by depolarizing steps from -70 mV consisted of the initial fast and subsequent sustained components. The former was inhibited by 10 mM nifedipine and Ca(2+)-free solution application whereas the latter decreased by only 16% under these conditions. Caffeine at 4 mM abolished the fast component and only slightly reduced the sustained component. Both component were inhibited by 1 mM TEA+. Adding 10 mM EGTA to the pipette solution abolished the fast outward current. In the presence of 10 mM TEA+ and 4 mM 4-AP an inward current was unmasked. These results suggest that the outward current in these cells is carried mainly via delayed rectifier and Ca(2+)-activated K(+)-channels. 4-AP inhibited the sustained component and had no effect on the initial fast outward current, thus A-current is apparently absent. PMID- 11424558 TI - [Clinical pathophysiology: current status, problems and future development]. PMID- 11424559 TI - [Effects of ionol, alpha-tocopherol and lipin on the processes of peroxidation in the syndrome of prolonged crush]. AB - Experimental investigations of the influence of ionol and alpha-tokoferol in combined usage and liposomes on the processes of lipids peroxidation in the heart and blood at the cellular and subcellular level during crunch syndrome were made. The injection of ionol and alfa-tokoferol combined and lipid decreases the production of lipids peroxidation and increase antioxidation organism status and as a result the survival abilities of animals. These data, refected antioxidation provision of the heart and blood under crunh syndrome at the cellular and subcellular levers are given. Low antioxidation provision of the heart must be taken in consideration, that grounds the necessity of taking into account of the antioxidation cellular defence modulations factors under the complex therapy working out. PMID- 11424560 TI - [The role of calcium-dependent mechanism in the realization of immune effects of kidney peptide complex]. AB - The role of intracellular and extracellular calcium in the realization of immune effects of the kidney peptide complex were studied. Expression of surface immunoglobulin receptors on lymphocytes depressed under the influence of calcium channels blockators (verapamil, phenigidin), binding of intracellular and extracellular calcium (EDTA, BAPTA). The kidney peptide complex restorted expression of surface immunoglobulins depressed by verapamil, phenigidin, EDTA, BAPTA. Also in this conditions the peptide complex increased the re-assembly of receptors, as caps, patches and clasters. Kidney peptide complex has a synergy with the calcium ionophor A 23187 to increase the expression of surface immunoglobulin receptor on lymphocytes. We could not exclude the direct influence of peptide complex on the conformation changes of the ion traffic systems for calcium. It was suggested that peptide complex promote the entrance of calcium ions into the cells. The influence of kidney peptide complex on expression of lymphocytes membrane receptors are independent from the calcium existence neither into cells nor outside the cells. PMID- 11424561 TI - [Changes of nitric oxide system during acute myocardial ischemic reperfusion]. AB - The reciprocal changes of NOS and arginase activity during acute myocardial ischaemia (90 min) and reperfusion (180 min) was shown in experiments on chest closed dogs with spontaneous breathing. NOS activity in the ischemia injured myocardial decreased on 60% while arginase activity increased on 487%. Levels of both alternative pathways of L-arginine metabolism altered reciprocally too. NO2( )-level was reduced on 57%, and urea level increased on 665%. The same changes were in arterial blood, started from 10 min of ischemia. These changes can play an important role for development of acute ischaemia treatment. PMID- 11424562 TI - [The influence of cortisol on several links of metabolism in myeloid cells and leukocytes]. AB - The influence of cortisol on the activity of enzymes that catalyze several stages of energy metabolism, proteolysis and antioxidant system in the bone marrow myeloid cells, neutrophils and lymphocytes of 7-10-day aged piglets was investigated. It was established that durable hormone administration (40 mg per kg body weight every 12 hours during 3 days) caused the decrease in the initial stages of glycolysis and pentose phosphate way intensities, as well as inhibition of cytochrome-c oxidase and antioxidant enzymes activities in the cells of mentioned populations. Such effects of cortisol on the intracellular metabolism may represent the important link in the mechanisms of glucocorticoid inhibition of myeloid cells and circulating leucocytes functional activities. PMID- 11424563 TI - [Dependence of bioelectrical activity in occipital and frontal lobes and of the function of visual pathways from blood and oxygen supply]. AB - The dependence of occipital and frontal lobes of the brain cortex bioelectrical activity and function from blood and oxygen supply was investigated in 626 minopic and 80 children and teenagers with normal vision. Myopia was found a natural model of blood and oxygen deprivation, of decrease of bioelectrical activity of frontal and partially occipital lobes, especially. The decrease of alpha-rythme index and alpha-waves amplitude was found on EEG of occipital lobes. Adaptation to hypoxia in the course of interval hypoxic training and the course of osteoreflex stimulation were proved to be reliable means for improving blood and oxygen supply of occipital and frontal lobes of the brain cortex, which was followed by normalization of EEG and vision: the sharpness of vision rose to 1.0 0.9 in 80% of children and in 60% of teenagers. PMID- 11424564 TI - [Modeling of K+ATP channel activity in normotensive and hypertensive animals]. AB - It was shown that vasodilatory effects of ATP-sensitive potassium channels opener -flocalin--depend from character of increase of initial vessel tone. Norepinephrine and angiotensin II partially inhibit effects of flocalin, thus they depressed of KATP channel activity. The inhibitory effect of angiotensin II increases under arterial hypertension conditions. It was shown that vasodilatory effects of flocalin significantly do not distinguish in experiments on normotensive and hypertensive rats under K(+)-depolarization conditions. While the vasodilatory effects acetylcholine on hypertensive rats are significantly diminished. These data suggests that flocalin could be a perspective substance for treatment of hypertension. PMID- 11424565 TI - [Sympathetic nervous system correlates of emotional tension in subjects with different status of the autonomic nervous system]. AB - 86 miners (age of 30-40) of coal miners were examined. It was ascertained that the level of tension of neurohumoral mechanisms of regulation of myocardium activity and splanchnic function of organism in emotiongenic conditions is determined by the condition of ANS. The secretion of noradrenaline of normotonics prevails, peripheral and central mechanisms of vegetative regulation of chronotropic function of heart start their work, tone of resistive vascular resis slightly. The discharge of adrenaline to blood of vago- and sympatotonic prevails, considerable tension of mechanisms of vegetative regulation of myocardium and expressed rise of tone resistive vascular and breathing frequency are observed. People with parasympatotonic are noted to have the biggest tension of splanchnic and regulative systems. The lower emotional steadiness is typical for them. PMID- 11424566 TI - [Immunological reactivity conditions of burned children]. AB - Analysis of the immunological reactivity conditions of burned children was conducted. The factors witch have influence on the functional activity of blood leucocytes were studied. The decrease of immunological reactivity proportionale to the grave of the trauma was discovered. The secondary immunodeficiency in T indepent type developed. The protein fraction of the blood serum influenced on the T-lymphocytes to the highest degree. Simultaneously decrease of the toleranse to the autologus erytrocytes with the hyperproduction of antibodies was discovered. PMID- 11424567 TI - [The dependence of blood and tissue proteolytic activity during acute stress from A type of the organism reaction]. AB - In the experiments on rats there was grounded the dependence of proteolytic processes in tissues during acute emotional painful stress from typological peculiarities of the organism that was defined by the method of "Open field". PMID- 11424568 TI - [Chronic administration of arginine-vasopressin stimulates insulin's synthesis in beta-cells of pancreas in intact and diabetic rats]. AB - Investigations on the intact and diabetic rats show us that chronic administration of [Arg8]-vasopressin for ten days (intracerebroventricular 24 pg or intraperitoneal 240 ng every day) leads to the increasing of insulin's level in beta-cells of the pancreas in normoglycemic rats to 6.% +/- 0.2% in comparison with the control group, and in hyperglycemic diabetic rats to 25.9% +/- 0.6% at the intracerebroventricular infusions, and to 15.9% +/- 0.8% at the intraperitoneal injections. The administrations of vasopressin in animals with experimental diabetes mellitus leads to the decreasing of glycemia's level in average to 37%, but do not have an influence on the destruction of Langerhans islets. Probably an insulin-stimulating effect of intracerebroventricular infusions of vasopressin is connected with the modulation of functional activity of hypothalamic centres of food behaviour and dorsal motor nucleus of vagal nerve and in peripheral injections vasopressin directly stimulates insulin synthesis. PMID- 11424569 TI - [Effect of sarcolemmal ion-transporting system blockers on the intensity of heart damage during "calcium paradox"]. AB - The aim of present study was to investigate a role of different anions in calcium paradox development. It is accepted point of view that development of calcium paradox is depend on cation composition and activity of Na/Ca exchange. However, role of anion composition remain unknown. It is not studied role of some aniontransporting systems in development of calcium paradox. Experiments were carried out on isolated Langendorff perfused rat hearts. Hearts were perfused with calcium-containing solution for 15 minutes, calcium-free medium for 10 minutes and reperfused by initial calcium-containing solution with [Ca2+ = 2 mM]. Release of myoglobin was used as a marker of membrane damage. It has been shown that addition of 5-20 mM HCO3 exacerbated calcium paradox of the heart, elevated myoglobin release from 4.92 +/- 0.57 mcg/g dry weight to 11.3 +/- 1.6 mcg/g dry weight. An inhibitor of HCO3/Cl exchange, 10 mcM L-644,711 depressed elevation of myoglobin release to 4.8 +/- 1.05 mcg/g dry weight. An inhibitor of Cl- channels, 5 mcM DIOA caused raising of myoglobin loss to 7.3 +/- 0.8 mcg/g dry weight during calcium paradox. These data show dependence of calcium paradox on anion composition. A possible reason for exacerbation of calcium paradox by HCO3- rich medium could be consistence of HCO3/Cl and Na/Ca exchange. The results discover new perspectives in myocardial protection of calcium overload. PMID- 11424570 TI - [Mechanisms of enhanced vascular contractility in experimental animals and humans in atherosclerosis]. AB - Contractile and electrical responses of muscle strips from rabbit aorta to 5-HT and hyper K solution were compared in controls group of rabbits fed a normal diet, and atherosclerotic animals fed 1% cholesterol diet for 12 weeks (atherosclerotic group). A study of vascular responses of human popliteal artery was also carried out. It was shown that smooth muscle cells from arteriosclerotic vessels of both rabbits and human exhibit an increased sensitivity to contractile action of hyperpotassium solution and 5-HT. We found no significant difference in the densities of L-type Ca2+ channel current as well as voltage dependent K+ channels current measured by patch clamp method in single smooth muscle cells from control and atherosclerotic rabbit aorta. Removal of endothelium from the vascular strips resulted in an enhanced vascular contractility in control but not in atherosclerotic group. The dose response curves for endothelium-free muscle strips for both 5-HT and hyper K solution were not significantly different between the two groups. Our result suggest that the enhanced vascular responses to 5-HT and hyper K under condition of arteriosclerosis could be attributed to an impaired function of endothelium which is a source of substances particularly nitric oxide that mediates vascular relaxation and provides a compensatory mechanism to keep peripheral resistance at lower level. PMID- 11424571 TI - [Autoimmune processes and biocompatibility in local damage of endothelium of the heart and its vessels]. AB - At immunoproductive processes in endothelium of patients with coronary atherosclerosis and endocarditis a phase dynamics of innate immunity represented by polytypic Ca(2+)-dependent autoprecipitating proteins of nonclonal recognition of surface cell membrane components (CMC) have been revealed. In case of coronary atherosclerosis the organism reaction to expressed CMC is manifested basically by intensified synthesis of cathodic complement-C3-similar autoprecipitin, but in case of endocarditis--by consumption of one with formation of anodic autoprecipitin and by intensified synthesis of autoprecipitating immunoglobulin G. These membranotropic autoimmune reactions are identical with the organism reactions to alien (heterologous) agents and differentially participate in development both of basic disease and perioperational complications after cardiac operations using trans- and implants, hence, they should be taken into account during membrane-stabilising and immunocorrecting therapy of recipients. PMID- 11424572 TI - Weimaraner warning. PMID- 11424573 TI - Congenital hypothyroidism in foals. PMID- 11424574 TI - An ethicist's commentary on giving an analgesic to mask pain in a horse. PMID- 11424575 TI - Is the veterinary profession losing its way? PMID- 11424576 TI - Principles of transfusion medicine in small animals. AB - The purpose of this review was to provide the reader with an updated overview of small animal transfusion medicine, and an approach to integrating it into private practice, based on a review of the veterinary and human literature spanning the last 3 decades. Electronic, online databases that were searched included CAB International and Medline; multiple keywords or subject headings were searched that were appropriate to each of the sections reviewed: canine and feline blood groups, blood-typing and crossmatching, donors, blood collection, storage, blood components, blood transfusion, blood component therapy, blood substitutes, and adverse reactions. The safe use of blood component therapy requires knowledge of blood groups and antibody prevalence, and knowledge of the means to minimize the risk of adverse reactions by including the use of proper donors and screening assays that facilitate detection of serological incompatibility. The 2 assays available to the practitioner are crossmatching, which is readily done in-house, and blood typing. Blood typing is available in the form of a commercial testing kit, through use of purchased reagents, or via a request to an external laboratory. The risk of potentially fatal adverse reactions is higher in cats than in dogs. The decision to transfuse and the type of product to administer depend on several factors, such as the type of anemia and the size of the animal. In conclusion, transfusion medicine has become more feasible in small animal practice, with improved access to blood products through either on-site donors, the purchase of blood bank products, external donor programs, or the availability of blood component substitutes. PMID- 11424577 TI - [Anesthesia of the New Zealand rabbit using the the combination of tiletamine zolazepam and ketamine-midazolam with or without xylazine]. AB - In this study, anesthesia levels obtained with tiletamine-zolazepam (TZ) and ketamine-midazolam (KM) with or without xylazine (X) were compared in rabbits. Reflexes (corneal, palpebral and withdrawal), blood parameters (PaO2, PaCO2, pH and ions HCO3-), cardiovascular function (heart rate and mean arterial blood pressure) and body temperature were evaluated before and after the injections of the anesthetic combination in the same rabbits (n = 10). With KM and TZ, no suppression of reflexes occurred. The body temperature and pH decreased and HCO3- increased similarly to KMX et TZX. Some physiological and blood parameters were less (PAM, PaCO2) and not (PaO2) affected comparatively to KMX et TZX. These protocols were of short duration of action and did not offer any anesthesia or analgesia. Therefore, their utilization should be restricted to short procedures where no painful manipulations are performed. Ketamine-midazolam-xylazine and tiletamine-zolazepam-xylazine on the other hand are indicated for interventions that require anesthesia. With these combinations, all reflexes were absent for 30 45 and 60-90 min following injections of KMX et TZX, respectively. However, these combinations induce cardiac depression, as well as a decrease of all measured blood parameters and body temperature and a reduction of PaO2. Supplementation with oxygen is recommended with the introduction of xylazine in the protocol. PMID- 11424578 TI - Lack of antibodies to porcine circovirus type 2 virus in beef and dairy cattle and horses in western Canada. AB - Porcine circovirus type 2 (PCV2) is a recently recognized agent that is consistently associated with postweaning multisystemic wasting disease in swine. There are conflicting data concerning the ability of this virus to infect and cause disease in other species. To determine if normal cattle, cattle affected with various illnesses, and normal horses in endemic areas of PCV2 infection in swine have had PCV2 infections, 100 randomly selected bovine sera, 100 equine sera, and 100 colostrum samples from clinically normal dairy cattle were examined for the presence of antibodies to porcine circoviruses by using ELISAs. All samples tested were negative for antibodies to porcine circoviruses. As well, a seronegative neonatal Holstein calf and 6 seronegative, 6-month-old beef calves that were experimentally infected with PCV2 failed to develop antibodies to the virus. These results suggest that natural infection of cattle and horses with PCV2 does not occur, or is a rare event, in western Canada. PMID- 11424579 TI - Peritonitis in a llama caused by Streptococcus equi subsp. zooepidemicus. AB - A 7-month-old, male llama was diagnosed with peritonitis caused by Streptococcus equi subsp. zooepidemicus. Clinical findings, medical treatment, and case outcome are described. Hematogenous dissemination from suspected pneumonia is proposed as the route of infection in this case. Possible transmission of the organism through contact with horses is discussed. PMID- 11424580 TI - Salmonella Give infection in 2 dairy herds. AB - Salmonella Give infection was investigated in 2 adjacent dairy herds because of the impact of milk contamination on public health. Once the status of the infection in a herd was established by fecal analyses, consecutive fecal testings, in conjunction with preventive measures, appeared to be consistent and cost-effective tools for the monitoring and control of salmonellosis in dairy herds. PMID- 11424581 TI - Reversed patent ductus arteriosus in a dog. AB - A 5-year-old cocker spaniel with tachycardia, > 2 s capillary refill time, arrhythmia, split S2 heart sound, right ventricular enlargement, flattened interventricular septum, dilated pulmonary artery, and slight tricuspid valve insufficiency was diagnosed with reversed patent ductus arteriosus (right-to-left vs the more common left-to-right shunt). Two years later, the dog was still stable. PMID- 11424582 TI - Noncompetition agreements--walk a mile in another's moccasins. PMID- 11424583 TI - New opportunities to optimize antimicrobial treatment. PMID- 11424584 TI - Challenges of managed care organizations in treating respiratory tract infections in an age of antibiotic resistance. AB - Managing respiratory tract infections (RTIs) presents many challenges to managed care organizations (MCOs). RTIs are among the most common illnesses treated by primary care clinicians; they seriously impact patient quality of life and are a leading cause of health-related absences from the workplace. The total direct costs of treating conditions such as acute otitis media, sinusitis, and acute exacerbations of chronic bronchitis are estimated to be $10.1 billion. The development of drug resistance has compounded these challenges by increasing treatment failures and costs and limiting treatment options. MCOs can meet these challenges by implementing clinical practice guidelines for acute respiratory infections, conducting focused studies of antibiotic use, and educating both healthcare clinicians and patients about appropriate antibiotic use. PMID- 11424585 TI - The crisis of resistant pathogens in respiratory tract infections--use of pharmacodynamic principles. AB - Infectious disease experts and public health officials continue to warn the medical community and the public that more strains of respiratory tract pathogens are becoming resistant to the antibiotics commonly used to eradicate them. The inappropriate use of antibiotics to treat viral infections has contributed to the development of multidrug resistance in the 3 key bacterial pathogens that cause respiratory tract infections: Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis. Traditionally, susceptibility of pathogens to antibiotics has been evaluated with in vitro testing by minimum inhibitory concentration (MIC) determination, which has also been used to establish breakpoints between susceptible and resistant organisms based on MIC distributions. However, a more clinical approach has been developed based on the correlation of pharmacokinetics (PK) and pharmacodynamics (PD) of antimicrobials with MICs and clinical studies, thereby establishing the new concept of PK/PD breakpoints. New guidelines for outpatient management of respiratory tract infections have been based on PD parameters. PMID- 11424586 TI - Appropriate antibiotic use in treating respiratory tract infections. AB - The emergence of drug-resistant pathogens, especially Streptococcus pneumoniae and Haemophilus influenzae, has complicated empiric treatment of both upper and lower respiratory tract infections. Clinicians are now forced to reevaluate their choices of first-line antibiotics. Although some bacterial respiratory infections may resolve spontaneously, the use of antibiotics has demonstrated a faster resolution of symptoms and prevention of sequelae and recurrences, thereby improving the patient's quality of life and ability to function. Therefore, it is recommended that all diagnosed bacterial respiratory infections be treated with an antibiotic. Factors that clinicians need to consider in prescribing an antibiotic are the predominant causative pathogens, rates of pathogen resistance, patient history, the bacteriologic and clinical efficacy, safety profile, dosing regimen, and cost effectiveness of available antibiotic choices. PMID- 11424587 TI - Clinical practice guidelines for the diagnosis and treatment of respiratory tract infections. AB - Clinical practice guidelines can be indispensable tools for managed care organizations (MCOs) in providing cost-effective treatment of common conditions. Guidelines for acute respiratory tract infections, such as acute otitis media (AOM) and acute sinusitis, can assist clinicians in accurately diagnosing these conditions, in providing treatment rationales, and in reducing the costs associated with inappropriate antibiotic prescriptions. Barriers to the implementation of practice guidelines include negative attitudes clinicians may have about guidelines promoted by MCOs; patient/parent expectations for antibiotic treatment; lack of financial resources, information system resources, and support for implementation; and lack of commitment to patient and provider education on the part of MCOs. MCOs can facilitate the adoption and implementation of guidelines with a systematic approach that involves establishing a guideline review process, gaining the support of providers, selecting outcomes measures, collecting and analyzing outcomes data, and providing feedback to clinicians about the impact of changes in their practices. This systematic approach should be used as part of the process for the National Committee for Quality Assurance accreditation. Evidence-based clinical practice guidelines for AOM and sinusitis have been developed recently by national consortia of infectious disease experts. Adoption of these guidelines can assist in preventing the spread of resistant pathogens. PMID- 11424588 TI - [The c-erbB-related oncoproteins in normal placenta and in gestational trophoblastic diseases (in vitro study)]. AB - In this study the expression of epidermal growth factor receptor (EGFR) and c erbB-2, c-erbB-3 and c-erbB-4 oncogenes were investigated in gestational trophoblastic diseases and normal first trimester placenta. Furthermore, the possibility that macrophage (IL-1 alpha, IL-1 beta, TNF) and lymphocyte (IL-2, gamma-IFN, GM-CSF) cytokines effects are mediated by changes in EGFR expression were studied. Paraffin sections of 16 cases of partial mole, 25 cases of complete mole, 10 cases of gestational choriocarcinoma and 11 cases of therapeutic abortion were studied immunohistochemically for EGFR, c-erbB-2, c-erbB-3 and c erbB-4 proteins. The presence of EGFR mRNA was studied using in situ hybridization. JEG-3 human choriocarcinoma cells were incubated with varying concentrations of interleukin 1-alpha, interleukin 1-beta, interleukin 2, gamma interferon, granulocyte-macrophage colony stimulating factor and tumor necrosis factor-alpha, and the expression of EGFR was measured by radioimmunoassay using a murine monoclonal antibody with specificity for EGFR. Staining for EGFR was detected immunohistochemically in all cell type in gestational trophoblastic diseases and normal placenta. The levels of expression of EGFR in choriocarcinoma and syncytiotrophoblasts and cytotrophoblasts in complete mole were significantly greater than those in syncytiotrophoblasts and cytotrophoblasts in both normal placenta and partial mole (p < 0.01, p < 0.01). The immunoreactivity of c-erbB-2 was significantly stronger in choriocarcinoma and extravillous trophoblast in complete mole than that in extravillous trophoblast in partial mole and normal placenta (p < 0.02, p < 0.01, respectively). Strong immunostaining for EGFR (p = 0.02) and c-erbB-3 (p < 0.01) in extravillous trophoblasts of complete mole was found to be significantly correlated with the development of persistent postmolar gestational trophoblastic tumor. Macrophage-derived cytokines IL-1 alpha, IL-1 beta and TNF significantly suppressed cell growth; this was associated with a significant increase in EGFR expression. The lymphocyte (IL-2, gamma-IFN, GM-CSF) cytokines had no significant effect on either EGFR expression or cell growth. These findings support the concept that cytokines may act as paracrine mediators of autocrine processes involved in choriocarcinoma cell growth regulation by modulating growth factor receptor expression. The EGFR-related family of oncogenes may be important in the pathogenesis and prognosis of gestational trophoblastic diseases. PMID- 11424589 TI - [Significance of contractile reserve in the assessment of cardiac diseases]. AB - One of the most important question in the revascularization of patients with impaired left ventricular function caused by coronary heart disease is the proper differentiation of necrotic myocardium and the region with the possibility of functional recovery. An improving in myocardial performance causes better survival in patients undergone revascularization. Preoperative assessment of segmental or global contractile reserve as a marker of myocardial viability by various methods in suitable to predict the improvement expected after procedure. It is useful in selection of the patients in whom the restoration of coronary blood flow takes a beneficial effect in their survival and quality of life. Authors based on relevant data summarize the methods in the evaluation of contractile reserve and its usefulness in clinical decision making of various heart diseases. PMID- 11424590 TI - [Suicidal behavior in the elderly. Review of results at the Pecs Center of the WHO/EURO Multicenter Study on Suicide]. AB - The authors examined the characteristics of old suicide attempters (over 65 years) with special regard to organic and depressive disorders and sociodemographic features. Within the framework of the WHO/EURO Multicentre Study on Parasuicide data of 849 suicide attempts were collected during three years. Comparing old age group (n = 54) to younger suicide attempters (n = 795) were found that over 65 years the proportion of female was much higher, repeaters rate was lower. In aged group were more widowed persons, while was not substantial difference in the rate of living alone. Regarding methods of parasuicide there were fewer poisoning (but higher rate of sedatohipnotics intoxication), while hard methods were more frequent. Over 65 years the proportion of depression, and organic disorders was higher. It seems that, in spite of the fact, that many old parasuicidal patients have depressive symptoms, they could get treatment only for symptoms of anxiety and sleep disturbance, while the affective problems remain undiagnosed and untreated. Another important factor, that high proportion of elderly sample was living in the family, with other relatives or in nursing home. In spite of this most of cases family members or caretakers in nursing home could not recognise the psychic problems and could not give sufficient emotional or social support to the patients to prevent suicide attempt. Authors pointed out, that the recognition and treatment of depression plays very important role in the suicide prevention in elderly population, and the adequate emotional and psychosocial support by family and health care systems seems to be essential. PMID- 11424592 TI - [Laszlo Lax, MD and his work in life insurance medicine]. PMID- 11424591 TI - [Fibro-osseous lesion of the central nervous system]. AB - The case of a 53-year-old woman with headache and progressive right sided decline of visual acuity is reported. Computed tomography scans of the brain revealed multiple circumscribed foci of mineralization located over the left frontal and parietal, as well as the right central brain parenchyma. Surgical sampling of the left frontal lesion yielded a conglomerate composed of mineralized vessels, myriad of psammoma bodies, and metaplastic lamellar bone entangled within poorly cellular collagen fibers. No evidence was found of an underlying vascular malformation or tumor, nor was there evidence of parenchymal necrosis of infectious origin. On account of the organoid association of the mesenchymal elements and the mineralized moiety, the lesion was consistent with fibro-osseous lesion of the central nervous system. Also known as "calcifying pseudotumor" of the brain, the origin of this exceedingly rare condition is, as yet, unknown. By analogy, its pathogenesis is likely to involve mechanisms underlying tumoral calcinosis of soft tissues. PMID- 11424593 TI - [Salomon Stricker (1834-1898)]. PMID- 11424594 TI - [The Markusovszky Hospital in Vas County: past, present and future perspectives]. PMID- 11424595 TI - [Defibrotide: a new antithrombotic agent]. PMID- 11424596 TI - [Forty years in the Referent Rubric. Reminiscences of a specialty correspondent]. PMID- 11424598 TI - [Medullary carcinoma of the thyroid gland]. AB - We report the experience of the Institute of Surgical Pathology of the University of Parma on three patients with sporadic medullary thyroid carcinoma (MTC). MTC is a tumor of parafollicolor cells origin (C cells). The surgical excision of the thyroid tumor and cervical node metastases is potentially curative. The other therapeutic options are limited. Considerable emphasis has been placed on early diagnosis and surgery for multiple endocrine neoplasia (MEN) related MTC. Genetic screening promises earlier and accurate diagnosis (RET gene mutations are found in MEN). PMID- 11424597 TI - [1900-2000. A century of Pediatrics in Parma]. PMID- 11424599 TI - [Surgical treatment of urologic complications of idiopathic retroperitoneal fibrosis. Our experience]. AB - The chief complications of retroperitoneal fibrosis RPF in the ureteric compression ab extrinseco by fibrotic tissue. In the period 1994 to 1999, we treated two cases of idiopathic RPF: the first case a 49 years aged man, presented a left hydronefrosis and bilateral ureteral displacement to the midline. RMN showed widespreads fibrotic tissue in the retroperitoneal space. At operation, the histological findings was that of aspecific fibrositis. The surgical procedure was ureterolysis and lateralization of both ureters with the aid of a wrapping with a pedicled omental graft. The patient was also treated with metilprednisolon, 8 mg-die, for altogether twelve months, Now he is asymptomatic. The second case, a man aged 60, had a compression of both ureter by fibrotic tissue down to the pelvic tract; the histologic picture was the same of that of the first case. A difficult ureterolysis was carried on, plus an omentoplasty and a left ureteral reimplantation to the bladder. Medical treatment followed, following the above scheme. Seven months after surgery the patient is well. This experience of ours urological complications of RPF is confirmatory of the importance of a sound surgical approach and of a tight follow up as well as a sustained corticosteroid therapy in order to prevent relapse. PMID- 11424600 TI - Hemangioma of the facial nerve. AB - To discuss the interest of the etiologic diagnosis of a facial palsy. STUDY DESIGN AND PATIENTS: Two cases of geniculate ganglion hemangioma with progressive facial palsy are reported. RESULTS: Hemangioma of the facial nerve is a rare and benign vascular tumor that originates from the venous plexus surrounding the facial nerve. The most common locations are the internal auditory canal and the geniculate ganglion. Diagnosis of these small tumors is radiological with CT-scan and MRI studies. Surgical excision through a supra-petrosal approach is the gold standard treatment. Hemangiomas of the facial nerve and particularly those developed in the geniculate ganglion area constitute a rare but not inconsiderable facial palsy etiology. An acute diagnosis and an early excision are fundamental to preserve a satisfactory facial function. PMID- 11424601 TI - [Unusual case of suicide with multiple cut injuries]. AB - A case is presented about the suicide committed by a 27-year-old male through multiple cut injuries in four distinct body regions (throat, left forearm, right wrist, right groin). The high number of injuries and the "non-typical" site of the lethal wound (groin) make this case relevant. PMID- 11424602 TI - [Pelvic floor rehabilitation as treatment of female urinary incontinence. Our experience]. AB - We refer herein on a simple program of rehabilitation that may be a first approach the treatment of, U.S.I. In our urodynamic service, we treated 38 female patients, affected by this condition, aged between 37-73 years. The same procedure was applied to all the patients, consisting of 10 seances twice weekly, during which we subjected the patients to biofeedback and vaginal electrostimulation of 50 Hz frequency. The results after 3 months were: 38% restored to normal, 51% improved, 11% unchanged; after 1 year were: 27% restored to normal, 49% improved, the remaining unchanged. As a whole, perineal rehabilitation by this technique can be curative in selected cases and, should it be necessary, can be repeated; its advantages are simplicity, low cost and lack of collateral effects. Motivation and compliance on the part of the patients are, anyhow, the chief factor of success. Moreover, the procedure does not preclude alternative treatments. It is suitable to chose cases where surgery is not so, or else is ill-accepted or has failed. PMID- 11424603 TI - [Report of 2 cases extragonadal germ cell neoplasia with primary burnt out tumor of the testis]. AB - If the histogenesis of the extragonadal germ cell tumor is a still debatable subject, its clinical diagnosis remains a question of no immediate solution. In fact, only the keen histologic evaluation of microfocuses and/or scar tissue in the testis, possibly on the guide of US finding, could give the answer about the primitiveness or not of the extragonadal neoplasia. Which implies, of course, some problems of compliance on the part of young locally symptomless men, especially on the ground of possibly bilateral involvement. PMID- 11424604 TI - Normal values of reactive oxygen metabolites on the cord-blood of full-term infants with a colorimetric method. AB - The main end-point of the study was to evaluate the normal values of reactive oxygen metabolites (ROMs) in healthy full-term babies. Secondary end-points were differences between groups related to modality of delivery, Apgar score, birth weight, gestational age and sex. All apparently healthy babies born at our institution between 8 a.m. and 8 p.m. Monday to Friday with gestational age 37-42 weeks, delivered both vaginally or by caesarean section and without foetal distress and perinatal asphyxia. ROMs were evaluated by a colorimetric method (d ROM test) on cord-blood immediately after birth. The values are reported as arbitrary unit U. Carr. Statistical analysis was performed by t-test and by multiple and stepwise regression analysis. We have analyzed 80 babies with mean birth weight 3301 +/- 446 g. and mean gestational age 39.5 +/- 1.0 weeks. The male:female ratio was 1.56 and the median (range) Apgar score was 9 (7-10) at 1' and 10 (9-10) at 5'. The babies born by vaginal delivery were 37 out of 80 while the remaining 43 were delivered by cesarean section. Because the two groups did not differ for the clinical characteristics they were considered together for the determination of the mean value of ROMs and indicated as "total". The mean value +/- SD of ROMs of the "total" was 115.5 +/- 32.6 U. Carr. Significant differences in the mean value of ROMs were not found related to type of delivery, birth weight, gestational age, and Apgar score at 1' and 5'. Instead the female infants had a significantly lower mean value of ROMs than the male babies (respectively 104.4 +/- 32.2 vs 120.2 +/- 30.6 U. Carr.; p = 0.031). Multiple and stepwise regression analyses both demonstrated that the sex of the neonate is able to independently influence the value of ROMs (respectively p = 0.025 and p = 0.035). The main end-point of the study was to determine the standard reference values for this method in the healthy full-term infant at birth: the values of ROMs we found in the "total" population are lower than those of healthy adults (between 250-300 U. Carr.) and similar to those of adults treated with steroids or antioxidant drugs. The finding that the female sex is able to independently determine lower values of ROMs at birth compared to the male sex, lets speculate that the female infants are less prone to oxidative stress in the first moments of life. PMID- 11424605 TI - [Use of initial high doses of growth hormone in the treatment of short children with deficient secretion of the hormone]. PMID- 11424606 TI - [New methods of percutaneous closure of patent ductus arteriosus]. AB - Various devices have been developed for the percutaneous closure of persistent patent ductus arteriosus (PDA). In recent years Gianturco coils have become increasingly widespread for this purpose. Poor control during positioning is a major disadvantage of Gianturco coils, with resultant embolization of coils and residual shunts. Very recently a new screwing detachable coil is available (Cook Cardiology, Europe). Between January 1996 and January 2000, 34 consecutive patients with PDA underwent cardiac catheterization. Thirty cases were judged suitable for this procedure, on the base of diameter and morphology of the ductus. Three cases were excluded because of the ductal diameter was too large (> 4.2 mm) and one because of its morphology (type B). In 28 patients a single coil was positioned, while in one case 2 coils were simultaneously used. In 29 cases (97%) complete closure was obtained (in 18 cases immediately, in 6 after 24 hours, in 5 after 1 month). One case showed a residual shunt and a second coil was applied eighteen months later with complete closure. So, considering also this last case, the closure was achieved in 100%. There were neither embolization or other complications. In conclusion, screwing detachable coil for PDA closure is a safe, very effective and at low cost technique. Therefore nowadays this procedure represents the method of choice for occlusion of small PDA (minimal diameter < or = 4 mm). PMID- 11424607 TI - Growth hormone insensitivity. PMID- 11424608 TI - Pediatric endocrinology and diabetology towards the new millennium. Expectations and limitations. PMID- 11424609 TI - [Digestive endoscopy in children]. AB - Also in the pediatric population the digestive endoscopy became, at this point, a very good diagnostic and operative technique largement used. The endoscopy can explore the upper gastrointestinal tract (esophagus, stomach, duodenum and jejunum) and can investigate the colon and the terminal ileum regarding the lower gastrointestinal tract. Principally, the operative endoscopy concerns emostasis of gastrointestinal bleeding, polipectomy, extraction of foreign bodies, and intestinal dilatations. The patients population submitted to this investigation by now is very wide thanks to the modernization of the instruments and at specialization of gastroenterologist pediatricians: range from newborn, to childhood, adolescence and young adults. The gastroenterologist pediatricians specialization associated to a routine use of amnemonic drugs (benzodiazepine) reduced at the minimum the invasivity of endoscopy; this technique can be considered a good safe procedure and free of important physical and psychic side effects for the young patient. PMID- 11424610 TI - [The pediatrician facing the problem of polluted air]. AB - The enormous amount of data concerning outdoor pollution has allowed quantifying the damage caused by single pollutants. On the basis of these data, and knowing the concentration of a pollutant in a given area, to date it is possible to foresee the expected risk for a given disease in our community. For instance, in an average size town like Parma, it can be calculated that in the middle of the winter, suspended particulate pollution (PM10), one day/three, will increase by 30% the number of symptomatic asthmatic children. In the long term, in a town like Parma, PM10 and NO2 pollution increases the number of children with more than 4 episodes of bronchitis or persistent cough by 30-50%. With regard to the typical summer pollution by O3, extremely harmful as it expands outside towns, in Parma and surroundings in August one should expect in 1/3 days double the number of symptomatic asthmatic patients, and every other day a 25% reduction of the respiratory function in asthmatic children exercising. With regard to indoor pollution besides the well known and severe problem of passive smoking, and that of volatile organic compounds still under investigation, one must consider NO2 pollution originating from the burning processes for heating and cooking. As this latter is extremely important in Italy, as gas is used in practically every house, the Author has done a specific research in which it is shown that if this kind of pollution was eliminated one could reduce the incidence of asthma in childhood from 7 to 5%, and noticeably the severity of the disease in affected subjects. Thus, paediatricians have two new aims. The first is to identify in pollution one of the possible causes of respiratory problems in each single patient, giving recommendations (ex. advise asthmatic children to avoid physical activity in the open and in the sunny hours during the summer, or avoid having children in the kitchen while cooking, or have boilers, heaters, etc. checked). Second, paediatricians should make aware, using the local actual data on pollution, not only the local administrators, but also and may be more important the parents of children with respiratory problems. These are in fact the ones who can and must become the most active in facing the big problem of pollution. PMID- 11424611 TI - [Thirty years of activity at the center for cystic fibrosis in Parma]. AB - One hundred fourteen patients (52 males, age range 1 to 40 yr, median age 16 yr) affected by Cystic Fibrosis (CF) are currently followed by the CF Centre of Parma. Forty two percent of them are adults. Most patients live in Emilia Romagna (59%) and Lombardia (19%). With respect to CF genotype, delta F508 mutation has been found in 54% of patients. Despite of the improvement in knowledge of CF, its diagnosis is still based on the clinical features and sweat test is usually used to confirm the diagnosis. Recent diagnostic tools, such as the genetic analysis and the potential difference measurement, could also be helpful in ambiguous situations. In the CF Centre of Parma, in agreement with the literature, the disease is now diagnosed earlier than in the past. Particularly, in the last year the median age of diagnosis was seven months, while it was 20 months in the 1980s. The early diagnosis and the improvement in the treatment of lung disease has significantly increased the median survival age of our patients, changing from 7.5 years in 1975 to 20 years in 1999. A significant improvement in CF prognosis could be further achieved by potential investigational agents, such as "protein-repair" addressed to CFTR activation. However, the gold standard for CF therapy is represented by gene therapy, although it is not yet available. PMID- 11424612 TI - Strategy for the use of vaccines in the next ten years. PMID- 11424613 TI - [From atopic eczema to respiratory allergy]. AB - The prevalence of atopic eczema in childhood is increasing. This is probably related with a deviation of immune system to Th2 directed to allergen instead of Th2 directed to microorganisms. Atopic eczema is genetically linked, but its expression is principally determined by environmental factors, allergens and infections. In children with atopic eczema, the immune system is altered. The number of Langheran's cells is augmented. Th2 cells infiltrate the skin and produce cytokines which mediate both immediate and late allergic reaction. Most of eczematous children is allergic to food. A IgE-mediated reaction take place at the mucosal intestinal site. The food challenge test is the most reliable mean to identify offending foods. Soy formula can be used in the treatment of cow milk allergy. The tolerance of formulae based on cow milk hydrolysed protein should be tested under medical supervision in children with cow milk allergy. Prevention of atopic disease with dietary interventions has not been successful. To prevent asthma, the administration of immunotherapy in children with allergic rhinitis seems to be promising. In eczematous children with elevated total IgE and specific IgE to inhalant allergens, cetarizine seems to prevent onset of asthmatic symptoms. PMID- 11424614 TI - [The pediatrician and antibiotic resistance]. PMID- 11424615 TI - [Mortality and morbidity in extremely premature newborns]. AB - The progress of perinatal medicine greatly reduced neonatal mortality and prognosys of the lowest gestational age (G.A.) newborns is improving continuously. The survival of the newborns with G.A. < 32 weeks doubled from the years 1973-1978 to 1995-99 in the our neonatal intensive therapy (TIN), whereas the incidence of the most serious pathologies of prematurity, which increases with the decreasing of G.A., has not changed in the years. In the surfactant era, Respiratory Distress Syndrome is not the first cause of death and Intra Ventricular Hemorrage and Retinopaty Of Prematurity occur now in the lower stage of severity in comparison with the past. Cortison profilaxys antepartum, delivery with caesarian section and agressive intensive neonatal therapy seem to be the reason of these results. PMID- 11424616 TI - [Neonatal emergency in the delivery room]. AB - Perinatal asphyxia is the main cause of morbidity and mortality in the new-borns and in the infants. During delivery the risk of severe asphyxia is high not only in pre-term infants but also in those at term. A correct assistance during the first minutes of life is crucial for the outcome of the asphyxiated babies. The interventions to perform are few and quite simple but they must be performed carefully. There are some standardised steps in the correct assistance of the new borns in the delivery room: A)--airways gentle cleaning. B)--breathing, maintain a good ventilation with manual assistance avoiding the use of oxygen. Oxygen is toxic not only for preterm infants but also for babies at term. C)--cardiac activity, check in order to maintain a good blood pressure. Permanent brain damages are related not only to the length and the severity of asphyxia but mainly to the cardiac activity and to the circulation., D)--drugs: only few drugs are seldom necessary for rescutation of the new-borns in the delivery room--beta mimetics, Na-bicarbonate, surfactant--and they must be used carefully. The time is another critical point to prevent permanent damages, in the severe asphyxia only few minutes and for this reason it is mandatory the presence in delivery room of a person with very good experience in the newborn resuscitation. PMID- 11424617 TI - [The pediatrician and the child in a multiethnic society]. AB - The increasing immigration from developing countries has determined new health problems. Italian Pediatricians must become acquainted with these health concerns such as the excessive use of public pediatric structures, especially hospitals, specific diseases unusual among Italian children (malaria, tuberculosis, tropical parassitosis, sickle cell anemia, rickets and others), control of previous immunizations and repetition when one is not sure of their efficacy. Moreover, immigrated children seem to be more susceptible to common pediatric diseases (mostly infections) than Italian children. Finally, children adopted from developing countries deserve special attention because they arrive from a very poor social context with frequent history of important infectious diseases, congenital diseases, malnutrition and abuse. Central precocious puberty is a frequent evolution in some of the adopted girls. PMID- 11424618 TI - [Treatment of thyroid dysfunction]. AB - The treatment of hypothyroidism and hyperthyroidism is briefly reviewed. L-T4, which is the treatment of choice of hypothyroidism, should be administered once daily after an overnight fast at the dosage of 10-12 mg/kg/day in newborns and of 100 mg/m2 in older children. FT4 and TSH serum levels will normalize within few weeks from the beginning of treatment and should be maintained in the normal range thereafter by appropriate increases of the dosage during growth. The first choice treatment in children with hyperthyroidism due to Basedow disease is the administration of antithyroid drugs, i.e. methimazole at the initial dosage of 0.4-0.6 mg/kg/day b.i.d. or t.i.d.; after remission, the dosage of this drug can either be reduced or L-T4 added in order to avoid hypothyroidism. Treatment should be continued for at least 24 months and the patient followed thereafter to detect possible relapses. Side effects are rare, the most frequent are skin rashes and the most severe is agranulocytosis, so complete blood count should be evaluated frequently especially in the first months of treatment. Surgery (total or near-total thyroidectomy) should be considered in case of failure of medical therapy, even if radioiodine administration, used until now mainly in adults, is gaining favour also for children especially in the United States. PMID- 11424619 TI - [Premature pubarche]. AB - Premature adrenarche refers to the appearance of public hair before age 8 years in girls and 9 years in boys, without other signs of puberty or virilization. Growth velocity may be increased and slightly advanced bone maturation is often present and is usually well correlated with the height age. The transient acceleraration of growth and of bone maturation have no negative effects on the onset and progression of puberty, and on final height. The diagnosis is based on the exclusion of the different forms of hyperandrogenism such as precocious puberty, late-onset congenital adrenal hyperplasia, virilizing tumors, glucocorticoid resistance, and Cushing syndrome. Once the diagnosis is made, no treatment is needed. However, a long-term follow-up of these patients is warranted. Recent data, in fact, indicate that girls with premature adrenarche may not have a benign outcome. Postpubertal girls with premature adrenarche during childhood have an increased frequency of functional ovarian hyperandrogenism. Furthermore, hyperinsulinemia is a common feature in adolescent patients with premature adrenarche and functional ovarian hyperandrogenism, and appears to be directly realted to the degree of androgen excess. Although the mechanisms interlinking the triad of premature pubarche, hyperinsulinemia and ovarian hyperandrogenism remain enigmatic, this frequent concurrence may result, at least in part, from a common early origin rather than from a direct interrelationship later in life. PMID- 11424620 TI - [Care recommendations for type 1 neurofibromatosis]. AB - Neurofibromatosis type 1 (NF1) is a progressive, multisystem disorder affecting about 1:3000 individuals. About one third of patients show serious complications and about one half are mildly affected. Since the original National Institutes of Health Consensus Conference in 1987, that established the clinical criteria for the diagnosis of NF1, there has been significant progress toward a more complete understanding of the molecular bases for NF1, and our knowledge of the natural history and management of the NF1 has significantly improved. Despite these advances, the diagnosis of NF1 is still based largely on clinical criteria and no individual prognostic evaluation or definitive medical therapy are available. The recommendations for the care of NF1 patients and their families are constantly changing: according to the new guidelines, the mainstay of management is anticipatory guidance and surveillance for treatable complications; surveillance usually includes annual follow-up visits, unless symptoms call for more frequent visits or more accurate diagnostic evaluation. PMID- 11424621 TI - [Genetics of type 1 neurofibromatosis]. AB - Neurofibromatosis type 1 (NF1) is an autosomal dominant disorder characterised by cafe au lait spots, multiple neurofibromas and Lisch nodules of the iris, with marked variability of expression. The NF1 gene is located at 17q11.2, spans 350 kb genomic DNA and comprises 60 exons encoding a 11-13 kb transcript (Viskochil et al.). Four alternatively spliced NF1 transcripts have been identified and they show differential expression in various tissues. NF1 gene is a member of the tumor suppressor gene family. The protein encoded by NF1, neurofibromin, has a domain homologous to the GTPase activating protein (GAP) family, and downregulates ras activity. Neurofibromin is involved in the control of cellular growth and differentiation and germline mutation analysis has shown that around 82% of all the fully characterised NF1 specific mutations so far predict severe truncation of neurofibromin. The current demand for molecular diagnosis of NF1 is low. Many couples would probably request a prenatal diagnosis if it could predict disease severity. Molecular prediction of disease severity and prognosis may either be very complicated or even impossible. Presymptomatic DNA diagnosis is probably not going to be in huge demand because the clinical diagnosis of NF1 is usually straightforward, even in early childhood. Further knowledge of the gene function may also lead to the development of new therapy for the disease. PMID- 11424622 TI - [Practical considerations on screening for microalbuminuria in children and adolescents with type 1 diabetes]. AB - Diabetic Nephropathy is one of the microvascular complications associated with type 1 diabetes mellitus with a major portion of the excess morbidity and mortality. In order to avoid or at least delay its onset we should detect a very small amount of proteins in the urine (between 15-20 and 200 micrograms/min.) that is a strong predictor of those likely to progress to overt nephropathy. This detection can be obtained evaluating an overnight sample of urine and suggest preventives interventions. such as intensive diabetes management and angiotensin converting enzyme inhibition (ACEi). Furthermore, associated risk factors should be prevented or treated (hypertension, retinopathy, hyperlipidemia and smoking). PMID- 11424623 TI - Standardized treatment cuts pneumonia deaths. AB - By developing a set of guidelines for admission parameters, antibiotic administration, and treatment, multidisciplinary health care professionals in a Utah health system have significantly reduced the number of deaths and hospital admissions for patients with community-acquired pneumonia. Lengths of stay also were reduced. Before the introduction of these standards, more than 70 different antibiotics had been used statewide. PMID- 11424624 TI - Improve patient safety to comply with new standards. PMID- 11424625 TI - How to define risk factors among cancer patients. PMID- 11424626 TI - 'Payer specialists' save nurses time. PMID- 11424627 TI - New collaborative tool helps case managers track diabetes patients. AB - For the first time, organizations representing physicians, health plans, and hospitals have collaborated in the development of a common set of evidence-based measures for evaluating performance in health care. Experts say case managers can use these new protocols to as a measuring stick for diabetes patients. The American Medical Association, the Joint Commission on Accreditation of Healthcare Organizations, and the National Committee for Quality Assurance have jointly established broadly applicable measures for the management of adult diabetes. The new tool, called the Coordinated Performance Measurement for the Management of Adult Diabetes, lays the groundwork for testing a single-source approach to measuring performance of care provided to diabetes patients in multiple settings. PMID- 11424628 TI - CM can play critical role in reducing medical errors. AB - These days, virtually every hospital in the country is actively engaged in finding ways to reduce medical errors. And now, many are catching on to the fact that case management can play a pivotal role in this pursuit. Indeed, the role of case managers in reducing medical errors is critical, says Michelle Gofney, director of case management at Deborah Heart and Lung Center, a 161-bed hospital with a full-service ambulatory care center in Mills, NJ. 'Case managers are the primary specialty voice to reduce medication errors,' explains Gofney. She says that is because case managers alone look at the patient's medical care through the physician's eyes and the nurse's eyes as well as every ancillary department in the hospital. PMID- 11424629 TI - Sodium fusidate (fusidin) ameliorates the course of monophasic experimental allergic encephalomyelitis in the Lewis rat. AB - We have evaluated the effect of the immunosuppressant sodium fusidate (fusidin) on the course of acute monophasic experimental encephalomyelitis (EAE) in male Lewis rats. Prophylactic treatment with fusidin, 80 or 120 mg/kg bd wt., markedly ameliorated the course of the disease in rats immunized with myelin basic proteins in complete Freund's adjuvant, entailing delayed onset of symptoms, lower clinical scores and more rapid recovery than PBS-treated control rats. The fusidin-treated, immunized rats exhibited milder mononuclear cell infiltration of brains and spinal cords than control animals. These data provide further evidence for the anti-inflammatory effect of fusidin and suggest that this drug may be valuable for the treatment of human multiple sclerosis. PMID- 11424630 TI - Day-to-day variability of maximum walking distance in MS patients can mislead to relevant changes in the Expanded Disability Status Scale (EDSS): average walking speed is a more constant parameter. AB - In this preliminary study we measured maximum walking distance and walking time on four consecutive days in 29 patients with clinically stable multiple sclerosis (MS). Patients were included in the study if they could achieve a maximum unaided walking distance of 100 up to 500 m. Our results showed a certain day-to-day variability of maximum walking distance, in some cases meaning changes up to 1.5 points in the expanded disability status scale (EDSS), which could be misinterpreted as a progression of the disease. Simultaneous measurements of maximum walking time showed a similar variability, unlike the mean walking speed which turned out to be more stable. Our results therefore suggest that scoring of MS patients should not be based on one single measurement of the maximum walking distance. The more reliable parameter appears to be the mean walking speed. PMID- 11424631 TI - An assessment of the spectrum of disability and handicap in multiple sclerosis: a population-based study. AB - OBJECTIVES: To establish the spectrum of disability and handicap in a population based sample of multiple sclerosis (MS) patients. BACKGROUND: Much knowledge exists about the epidemiology of MS but, despite its importance for health and social service planning, there remains relatively little data on the extent and nature of disability and handicap in this population. METHODS: In a prevalence study in the north-east of N. Ireland, 288 patients (Poser criteria) were identified. Disability and handicap were assessed using the Incapacity Status Scale and Environmental Status Scale of the Minimal Record of Disability for MS. RESULTS: Both scales were completed for 248 (86%) of patients. Just 71 (29%) are fully independent in all basic ADL's of bathing, dressing, grooming and feeding. Fifty-seven (23%) are unable to climb a flight of stairs and 102 (42%) acknowledge problems with sexual function. Sixty-one (25%) were working essentially full-time and 53 (21%) had no external financial support. Forty-five (18%) had changed residence due to MS, 12 (5%) were institutionalised and 86 (35%) required assistance for at least 1 h/day with ADL's. Eighty-one (33%) were unable to drive a car or use public transport. Forty-two (17%) access community services for at least 1 h/day on average. CONCLUSIONS: This data gives a clear indication of the considerable range of basic health and social issues in a typical MS community. Further work is required to establish patient perceptions of the adequacy of care provision and whether standards of care for MS patients are being met. PMID- 11424632 TI - Disease specific quality of life instruments in multiple sclerosis: validation of the Hamburg Quality of Life Questionnaire in Multiple Sclerosis (HAQUAMS). AB - Quality of life (QoL) is discussed as an additional outcome measure in multiple sclerosis (MS). However, few questionnaires assessing disease specific QoL in MS have been published. On the basis of the literature and interviews with clinicians and MS patients, we have developed a disease specific QoL instrument and validated it in a broad range of patients with MS. In this study, a heterogeneous sample of n = 237 MS patients completed the newly developed Hamburg Quality of Life Questionnaire in Multiple Sclerosis (HAQUAMS, in German language) and a battery of already validated questionnaires. They further underwent neurological scoring and objective tests. By these means, we investigated its validity, appropriateness, internal consistency, and retest reliability. Internal consistency and retest coefficients were high and satisfied psychometric standards. Convergent and discriminant validity was supported by direction, magnitude and pattern of correlations with other health measures. HAQUAMS subscales and its total score distinguished between patient groups of varied disease severity, cognitive impairment, and affective symptomatology. No floor or ceiling effects were found in either of the HAQUAMS subscales. The HAQUAMS is a reliable, valid and appropriate instrument for QoL assessment in multiple sclerosis. Data of responsiveness are currently being obtained. PMID- 11424633 TI - The relation between objective and subjective impairment in cognitive function among multiple sclerosis patients--the role of depression. AB - OBJECTIVE: To evaluate the relations between perceived cognitive function and objective cognitive deficit and to assess variables affecting perceived cognitive function among multiple sclerosis (MS) patients. METHODS: A cross sectional study of patients with MS. All patients were interviewed and the Expanded Disability Status Scale (EDSS) score was determined. The dependent variables were four items assessing perceived concentration and thinking, attention, memory, and whether others have noticed memory or concentration problems. The explanatory variables were age, sex, duration of disease, number of relapses in the last 2 years, EDSS score, depressive symptoms score (CES-D) and the domains of the Neurobehavioral Cognitive Status Examination (NCSE) assessing cognitive performance. Bivariate and then multivariate analysis were performed. RESULTS: One hundred and sixty-one MS patients were included. Mean age was 44.2 years (s.d. 11.3 years), mean EDSS score was 4.86 (s.d. 1.93). Seventy-two per cent of the patients had objective cognitive impairment and 51% reported decreased perceived cognitive function. In all models assessing perceived cognitive function we could explain only a small part of the variance (R2 ranged between 18-26%). In all these models depressive symptoms explained the highest portion of the variance (partial R2 ranging between 13-26%). The only domain of the NCSE that entered some of the models was calculation (partial R2 ranging between 3-7%). CONCLUSIONS: These findings emphasize the gap between objective and subjective assessment of cognitive function and the high correlation between perceived cognitive deficit and depressive symptoms. PMID- 11424634 TI - Recovery following acute exacerbations of multiple sclerosis: from impairment to quality of life. AB - To observe the pattern of recovery after treatment with intravenous Methylprednisolone (i.v. MP) for a relapse of multiple sclerosis (MS), and to determine the best time to plan further interventions such as rehabilitation, we assessed consecutive outpatients (n = 24) treated with i.v. MP for a relapse over a period of 12 weeks. Outcomes measures used were the Expanded Disability Status Scale (EDSS), the Incapacity Status Scale (ISS), the MOS Short Form-36 (SF-36), the Mental Health Inventory (MHI), and the MS-Related Symptom Checklist (MSSCL). There was statistically significant early improvement of EDSS and ISS scores, which was sustained until week 12, and significant improvement of MHI and MSSCL scores between 4 and 12 weeks. Although trends for improvement of scores reflecting the same pattern of recovery were observed with the SF-36 physical and mental composites, these changes did not reach statistical significance. Our results suggest that improvement of impairments and disability after treatment with i.v. MP for a relapse of MS occurs early, while improvement of subjective health status is delayed. Even after maximum improvement is reached, patients are left with multiple symptoms and functional limitations, and may benefit from additional rehabilitative interventions. PMID- 11424635 TI - Quantitative contrast-enhanced magnetic resonance imaging to evaluate blood-brain barrier integrity in multiple sclerosis: a preliminary study. AB - Gadolinium enhanced magnetic resonance imaging detects focal blood-brain barrier breakdown in new inflammatory multiple sclerosis lesions, but such lesions do not correlate with disease progression. To explore whether the latter might relate to subtle but widespread blood-brain barrier (BBB) breakdown with low grade inflammation mediating tissue damage, quantitative techniques were used to detect subtle gadolinium enhancement within otherwise normal-appearing white matter and within lesions not showing visible enhancement. T1-weighted imaging was performed prior to and at 5, 20 and 40 min following injection of 0.3 mmol/kg gadopentate dimeglumine in 33 patients with multiple sclerosis and five healthy control subjects. In healthy controls, a significant increase in white matter signal 5 min following contrast injection was observed (1.8%, P < 0.0005); the signal returned to baseline values by 20 min. In multiple sclerosis patients, a non significant trend was noted for signal to remain elevated in normal-appearing white matter at the 20 and 40 min post-contrast time points; this was most apparent in primary progressive multiple sclerosis. Significant increases in signal intensity were noted at all time points post contrast in apparent non enhancing lesions. The transient post contrast signal increase in controls is likely due to intravascular gadopentate dimeglumine. The persistent increases in signal intensity in non-enhancing lesions suggest more widespread abnormalities in BBB than is visually apparent, but substantiation of BBB leakage in normal appearing white matter will require further study using more sensitive methods. PMID- 11424636 TI - Water diffusion is elevated in widespread regions of normal-appearing white matter in multiple sclerosis and correlates with diffusion in focal lesions. AB - Pathological changes in the normal-appearing white matter in multiple sclerosis are well recognised, but their relationship to pathology in focal lesions is not well understood. Magnetic resonance diffusion imaging is sensitive to abnormalities in the integrity, size and geometry of water spaces in brain tissue. This study investigated the anatomical distribution of normal-appearing white matter diffusion abnormalities and their relationship to diffusion in focal lesions in multiple sclerosis (MS). The average apparent diffusion coefficient (ADCav) was measured by three-axis echoplanar diffusion imaging in normal appearing white matter regions and lesions throughout the brain in 40 patients, and in white matter in 14 matched controls. The correlation between the ADCav in normal-appearing white matter and lesions was determined. In controls and patients, diffusion was highest in the corpus callosum. Patients had a higher mean ADCav than controls in widespread regions including the corpus callosum, cerebellar, temporal and occipital normal-appearing white matter. Mean normal appearing white matter ADCav correlated strongly with mean lesion ADCav (r = 0.67, P < 0.001). This study demonstrates that water diffusion is elevated in widespread areas of normal-appearing white matter in MS, and is correlated with diffusion in lesions. These findings suggest that the pathogenetic mechanisms causing tissue damage in lesions and normal-appearing white matter are at least partly linked. PMID- 11424637 TI - Additive effect of the HLA-DR15 haplotype on susceptibility to multiple sclerosis. AB - Multiple sclerosis (MS) has been associated with the human leukocyte antigen DR15 allele in Caucasians of North and Central European origin. However, the relative effect of the DR15 homozygous and the DR15 heterozygous genotypes on the disease susceptibility is unclear. Based upon results from three North European studies we have examined this by meta-analysis. Our results suggested that the effect of the DRB1*1501,DQA1*0102,DQB1*0602 haplotype on the susceptibility to MS is additive, perhaps reflecting that development of the disease is facilitated by a high density surface expression of the antigen presenting molecules encoded by this haplotype. Possible implications of our finding to future studies of the genetic background of MS is discussed. PMID- 11424638 TI - Dendritic cells derived from patients with multiple sclerosis show high CD1a and low CD86 expression. AB - Dendritic cells (DC) are important antigen presenting cells (APC) and play a major role in initiating and orchestrating immune responses by priming T cells. Little is known about involvement of DC in multiple sclerosis (MS), where auto aggressive T cells against myelin autoantigens are considered to contribute to inflammation and demyelination in the central nervous system. In this study, we compared phenotype and cytokine secretion of DC from patients with MS, other neurological diseases (OND) and healthy subjects. DC were generated from blood adherent mononuclear cells (MNC) by culture for 7 days with granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). The yield and morphology of DC were similar in MS patients and controls. In both, the DC phenotype was that of immature myeloid lineage, comprising CD1a+ and CD11c+. The proportion of CD1a+ DC, being important for presentation of lipid antigens to T cells, was higher in MS patients compared to controls. The proportion of CD86+ DC, a co-stimulatory molecule that is assumed to promote Th2 differentiation, was low in MS. Low proportions of CD86+ DC were only observed in untreated MS patients but not in patients treated with IFN-beta. Production of IL-10 and IL-12 p40 by DC did not differ in MS patients and controls. These findings indicate that alterations of functionally important surface molecules on DC are associated with MS. PMID- 11424639 TI - Ethnic minority health. Restricted access. AB - A survey of health authorities and primary care groups with high ethnic minority populations in England showed HAs were further advanced in addressing the special needs of ethnic minorities than PCGs. More than three-quarters of HAs cited initiatives for ethnic minorities compared with only 36 per cent of PCGs. Only a minority of organisations reported schemes to address health inequalities. Commissioning organisations covering large communities of people from ethnic minorities should do more to address their special health needs. PMID- 11424640 TI - In the morale maze. PMID- 11424641 TI - Film locations. Set for action. PMID- 11424642 TI - A confluence of contexts: asymmetric versus global failures of selective attention to stroop dimensions. AB - In 6 experiments probing selective attention through Stroop classification, 4 factors of context were manipulated: (a) psychophysical context, the distinctiveness of values along the color and word dimensions; (b) set size context, the number of stimulus values tested; (c) production context, the mode used to respond; and (d) covariate context, the correlation between the dimensions. The psychophysical and production contexts mainly caused an asymmetry in selective attention failure between colors and words, whereas the set size and covariate contexts contributed primarily to the average or global magnitudes of attentional disruption across dimensions. The results suggest that (a) Stroop dimensions are perceptually separable, (b) J.R. Stroop's (1935) classic findings arose from his particular combination of contexts, and (c) stimulus uncertainty and dimensional imbalance are the primary sources of task and congruity effects in the Stroop paradigm. PMID- 11424643 TI - Illusory conjunctions are alive and well: a reply to Donk (1999). AB - When presented with a red T and a green O, observers occasionally make conjunction responses and indicate that they saw a green T. These errors have been interpreted as reflecting separable processing stages of feature detection and integration with the illusory conjunctions arising from a failure at the integration stage. Recently, M. Donk (1999) asserted that the phenomenon of illusory conjunctions is an artifact. Conjunction reports are actually the result of confusing a nontarget item (O in the example above) for a target item (the letter T) and (correctly) reporting the color associated with the (incorrectly) selected target. The authors demonstrate that although target-nontarget confusion errors are a potential source of conjunction reports, there is a plethora of findings that cannot be accounted for by this confusion model. A review of the literature indicates that in many studies, illusory conjunctions do result from a failure to properly integrate features. PMID- 11424644 TI - Illusory conjunctions die hard: a reply to Prinzmetal, Diedrichsen, and Ivry (2001). AB - M. Donk (1999) showed that various data patterns that have been considered as evidence for the existence of illusory conjunctions may be due to errors of target-nontarget confusion, an account that challenges the mere existence of illusory conjunction. In a reply, W. Prinzmetal, J. Diedrichsen, and R. B. Ivry (2001) argued against this conclusion, claiming that some earlier findings can be explained only when one assumes that illusory conjunctions exist. The current article shows that Prinzmetal et al.'s claims cannot refute any of Donk's earlier conclusions, suggesting indeed that one can only conclude that "illusory conjunctions are an illusion." PMID- 11424645 TI - Pseudohomophone effects in lexical decision: still a challenge for current word recognition models. AB - Computational models that implement a serial mechanism of phonological assembly predict interactions between the size of the pseudohomophone (PsH) effect and stimulus length. Models with frequency-sensitive word representations predict baseword frequency effects. These predictions were tested in a lexical-decision task. The results showed constant PsH effects across different word lengths (in favor of parallel phonological activation) and baseword frequency effects (in favor of frequency-sensitive representations). However, the baseword frequency effect was opposite of what the models predicted. This result is most easily accommodated by models that assume an orthographic verification mechanism. The plausibility of such a mechanism was further supported by the results of 2 additional experiments investigating the effects of response speed and spelling probability (feedback consistency) on the size of the PsH effect. PMID- 11424646 TI - Dynamic illusion effects in a reaching task: evidence for separate visual representations in the planning and control of reaching. AB - The effects of an orientation illusion on perception and 2 different actions were investigated. An 8-cm x 2-cm cylindrical bar was placed in front of participants at various orientations. A background grating was used to induce an orientation illusion. In a perception task, the illusion affected participants' ability to align the bar with their sagittal planes. In one reaching task, a similar effect of the illusion was found on the choice between 2 possible grasping postures. In a second reaching task involving a single grasping posture, the orientation illusion affected the orientation of the hand at the beginning of the reach but not near its end. The authors argue that reaching trajectories are planned and initiated through a context-dependent representation but are corrected on-line through a context-independent representation. The relation of this model to a more general dichotomy between perception and action is discussed. PMID- 11424647 TI - Categorical perception of face identity in noise isolates configural processing. AB - Neuropsychological evidence suggests that face recognition based on configural (holistic) information can occur in isolation from recognition based on local feature cues. The present study shows that configural processing can be isolated experimentally in normal subjects. A phenomenon is reported that exists only for upright whole faces, namely categorical perception (CP) of face identity in noise. Three discrimination tasks (ABX, better likeness, and similarity ratings) were used to test for perceptual distortion across the category boundary predicted from binary classification of face morphs. Noise was added such that any single local region provided unreliable cues to identity. Under these conditions, CP was found for upright faces but not for inverted faces or single features, even with more than 10,000 trials. The CP-in-noise signature phenomenon was then used to show that configural processing survives image plane rotations of 45 degrees-90 degrees. PMID- 11424648 TI - Phase correction, phase resetting, and phase shifts after subliminal timing perturbations in sensorimotor synchronization. AB - Recent studies of synchronized finger tapping have shown that perceptually subliminal phase shifts in an auditory sequence are rapidly compensated for in the motor activity (B. H. Repp, 2000a). Experiment 1 used a continuation-tapping task to confirm that this compensation is indeed a phase correction, not an adjustment of the central timekeeper period. Experiments 2-5 revealed that this phase correction occurs even when there is no ordinary sensorimotor asynchrony- when the finger taps are in antiphase or arbitrary phase relative to the auditory sequence (Experiments 2 and 3) or when the tap coinciding with the sequence phase shift is withheld (Experiments 4 and 5). The phase correction observed in the latter conditions was instantaneous, which suggests that phase resetting occurs when the motor activity is discontinuous. A prolonged phase shift suggestive of overcompensation was observed in some conditions, which poses a challenge to pure phase correction models. PMID- 11424649 TI - A paradoxical exposure-duration effect in the Stroop task: temporal segregation between stimulus attributes facilitates selection. AB - In comparison with the situation in which target and distractor are presented until response, Stroop interference substantially decreases when the target color is removed from the display 150-160 ms after stimulus onset (W. La Heij, N. A. Kaptein, A. C. Kalff, & L. de Lange, 1995; O. Neumann, 1986). Six experiments showed that this paradoxical exposure-duration effect is robust, that it is not due to the target's exposure duration per se, and that it is not due to an increased spatial selectivity of input selection. The present findings support and extend earlier observations that changes in the display during stimulus exposure affect attentional selection. The findings are tentatively interpreted in terms of (a) reduced duration of input selection or (b) facilitation of attribute selection. PMID- 11424650 TI - The figure has a shape, but the ground does not: evidence from a priming paradigm. AB - In four experiments, the authors examined the extent to which the ground interpretation of an edge may receive a shape description. These experiments used the priming effect that shapes have on perceptual judgments on a subsequent trial. A robust reduction in error rates and reaction times was seen when the figural shape was the same as that on the previous trial. This repetition priming effect may be due to activation of the shape description of the figure that remained from the previous trial. In contrast, no priming by the shape of the ground was seen even when the contrast sign of the figure reversed between trials. Priming for figural shapes occurred at a relatively abstract level because it was robust across reversals of contrast and orientation. These data suggest that the figural interpretation of a shape receives a shape description but that the ground does not. PMID- 11424651 TI - Do infants segment words or recurring contiguous patterns? AB - Eight experiments tested the hypothesis that infants' word segmentation abilities are reducible to familiar sound-pattern parsing regardless of actual word boundaries. This hypothesis was disconfirmed in experiments using the headturn preference procedure: 8.5-month-olds did not mis-segment a consonant-vowel consonant (CVC) word (e.g., dice) from passages containing the corresponding phonemic pattern across a word boundary (C#VC#; "cold ice"), but they segmented it when the word was really present ("roll dice"). However, they did not segment the real vowel-consonant (VC) word (ice in "cold ice") until 16 months. Yet, at that age, they still did not false alarm on the straddling CVC word. Thus, infants do not simply respond to recurring phonemic patterns. Instead, they are sensitive to both acoustic and allophonic cues to word boundaries. Moreover, there is a sizable developmental gap between consonant- and vowel-initial word segmentation. PMID- 11424652 TI - Missing information in spoken word recognition: nonreleased stop consonants. AB - Cross-modal semantic priming and phoneme monitoring experiments investigated processing of word-final nonreleased stop consonants (e.g., kit may be pronounced /kit/ or /ki/), which are common phonological variants in American English. Both voiced /d/ and voiceless /t/ segments were presented in release and no-release versions. A cross-modal semantic priming task (Experiment 1) showed comparable priming for /d/ and /t/ versions. A second set of stimuli ending in /s/ were presented as intact, missing /s/, or with a mismatching final segment and showed significant but reduced priming for the latter two conditions. Experiment 2 showed that phoneme monitoring reaction time for release and no-release words and onset mismatching stimuli (derived pseudowords) increased as acoustic-phonetic similarity to the intended word decreased. The results suggest that spoken word recognition does not require special mechanisms for processing no-release variants. Rather, the results can be accounted for by means of existing assumptions concerning probabilistic activation that is based on partial activation. PMID- 11424653 TI - The influence of temporal selection on spatial selection and distractor interference: an attentional blink study. AB - Both spatial and temporal selection require focused attention. The authors examine how temporal attention affects spatial selection. In a dual-task rapid serial visual presentation paradigm, temporal selection of a target (T1) impairs processing of a second target (T2) that follows T1 within 500 ms. This process is the attentional blink (AB). To test the effects of withdrawing temporal attention, the authors measured concurrent distractor interference on T2 when the distractors were presented during and outside of the AB. Perceptual interference was manipulated by the similarity in color between T2 and concurrent distractors, and response interference was manipulated by the flanker congruency task. Results showed that perceptual interference was larger during the AB. Response interference also increased during the AB, but only when perceptual interference was high. The authors conclude that temporal selection and spatial selection rely on a common attentional process. PMID- 11424654 TI - Morphological facilitation following prefixed but not suffixed primes: lexical architecture or modality-specific processes? AB - Morphological facilitation was examined in immediate (Experiment 1) and long-term (Experiment 2) lexical decision with English materials. For the target (payment), related primes consisted of base-alone (pay), affix-plus-base (prepay), or base plus-affix (payable) combinations, thereby defining position of overlap. In addition, modality of presentation varied for primes and targets (Experiment 1). At short lags, the advantage for prepay-payment over payable-payment type pairs was significant when primes were visual (V) and targets were auditory (A), marginal under AV conditions, and nonexistent under VV conditions. At long lags, the magnitude of VV did not vary with position of overlap. Morphological facilitation was stable across changes in modality following prefixed and simple forms, reflecting lexical architecture. By contrast, the absence of facilitation following suffixed primes presented cross-modally implicates modality-specific processing. PMID- 11424655 TI - Visual localization of the center of mass of compact, asymmetric, two-dimensional shapes. AB - This study aimed at understanding how visual information is used to locate the center of mass. The center of mass is an important physical property of objects that must be taken into account when grasping and/or manipulating them. Participants were instructed to identify the point of equilibrium of compact, bidimensional, massless shapes displayed on a touch screen. The point of equilibrium was defined as the point on the face of the object that would allow one to balance the object in the horizontal position. Seven different triangles and 18 different quadrilaterals in different orientations were used as stimuli. It was found that participants can accurately and consistently estimate the position of the center of mass. The small observed errors were systematically influenced by the shape of the object. The participants tended to locate the center of mass at the center of an inscribed circle instead of the true center of mass. In general, the shape effect was impervious to the orientation of the figure and to the mode of response (left hand, right hand, or mouse). PMID- 11424656 TI - Visual search for motion-form conjunctions: is form discriminated within the motion system? AB - Motion-form conjunction search can be more efficient when the target is moving (a moving 45 degrees tilted line among moving vertical and stationary 45 degrees tilted lines) rather than stationary. This asymmetry may be due to aspects of form being discriminated within a motion system representing only moving items, whereas discrimination of stationary items relies on a static form system (J. Driver & P. McLeod, 1992). Alternatively, it may be due to search exploiting differential motion velocity and direction signals generated by the moving-target and distractor lines. To decide between these alternatives, 4 experiments systematically varied the motion-signal information conveyed by the moving target and distractors while keeping their form difference salient. Moving-target search was found to be facilitated only when differential motion-signal information was available. Thus, there is no need to assume that form is discriminated within the motion system. PMID- 11424657 TI - Superior visual search in autism. AB - Children with a diagnosis of autism and normally developing children, matched for age and general ability, were tested on a series of visual search tasks in 2 separate experiments. The children with autism performed better than the normally developing children on difficult visual search tasks. This result occurred regardless of whether the target was uniquely defined by a single feature or a conjunction of features, as long as ceiling effects did not mask the difference. Superior visual search performance in autism can be seen as analogous to other reports of enhanced unique item detection in autism. Unique item detection in autism is discussed in the light of mechanisms proposed to be involved in normal visual search performance. PMID- 11424658 TI - Validity and boundary conditions of automatic response activation in the Simon task. AB - Three experiments were conducted to determine whether spatial stimulus-response compatibility effects are caused by automatic response activation by stimulus properties or by interference between codes during translation of stimulus into response coordinates. The main evidence against activation has been that in a Simon task with hands crossed, responses are faster at the response location ipsilateral to the stimulus though manipulated by the hand contralateral to the stimulus. The experiments were conducted with hands in standard and in crossed positions and electroencephalogram measures showed coactivation of the motor cortex induced by stimulus position primarily during standard hand positions with visual stimuli. Only in this condition did the Simon effect decay with longer response times. The visual Simon effect appeared to be due to specific mechanisms of visuomotor information transmission that are not responsible for the effects obtained with crossed hands or auditory stimuli. PMID- 11424659 TI - [Disturbances in left ventricular diastolic filling. Pathophysiology and clinical significance]. PMID- 11424660 TI - [Elevated cortisol level in blood of patients with unstable coronary disease and acute myocardial infarction--new approach]. AB - The aim of this study was to assess changes in serum cortisol concentration, enzymatic markers of myocardial ischemia and necrosis, leucocytosis and glucose in patients with unstable coronary disease and acute myocardial infarction. The study was performed in 75 patients (20 females and 55 males aged 38-75 years; mean age 59.4 +/- 6.52), including 13 patients with unstable coronary disease (6 females and 7 males aged 40-66 years; mean age 59.4 +/- 6.52; group I), 40 patients with acute myocardial infarction (8 females and 32 males aged 38-72 years; mean age 57.3 +/- 9.57; group II) and 22 healthy volunteers (6 females and 16 males aged 39-75 years; mean age 53.1 +/- 9.62; control group). Acute ischemia as well as myocardial infarction are potent stress factors that destabilisate the functional equilibrium of the body. Considering the mechanism of action of cortisol and its physiological role, it seems that current views on elevated plasma cortisol levels as a response to stress and pain in infarction and acute ischemia should be supplemented. Anti-inflammatory properties of cortisol deserve more attention, while elevated levels might be of prognostic value in the above mentioned diseases. PMID- 11424661 TI - [Which medical treatment can minimize late complications after pharmacologic treatment of unstable angina?]. AB - Current pharmacological treatment of unstable coronary diseases has significantly reduced the rate of early and late complications. Aim of this study was to investigate the impact of in-hospital and ambulatory treatment on the occurrence of late complications in patients with unstable angina treated conservatively. The study group consisted of consecutive 79 patients hospitalized in our Department on the turn of years 1998-1999 (48 men--age 62 +/- 11 years), followed up by at least 1 year. We analyzed the administered agents on the first day of hospitalization: acetylsalicylic acid, heparin, nitroglycerin, beta-adrenergic agents, calcium channels blockers, statins and ACE inhibitors, as well these drugs prescribed for ambulatory management. At least one complication (recurrent hospitalization due to coronary artery disease, unstable angina pectoris, myocardial infarction, need for revascularization or death) occurred in 57 patients (72%) in a follow-up of 16 +/- 3 months. The following drugs significantly reduced rate of late complications (OR--odds ratio, CI--confidence interval, p--significance level):--intravenous heparin during hospitalization: OR = 0.22; CI = 0.12-0.42; p = 0.007;--acetylsalicylic acid administration during hospitalization: OR = 0.42; CI = 0.22-0.77; p = 0.03;--acetylsalicylic acid prescribed for ambulatory treatment: OR = 0.24; CI = 0.13-0.46; p = 0.05. CONCLUSIONS: Heparin and acetylsalicylic acid treatment seems to reduce the number of late complications in patients treated conservatively for the episode of unstable angina. PMID- 11424662 TI - [Normalization of lipid metabolism disturbances and clinical course after percutaneous coronary angioplasty: 6 month observation]. AB - The recurrent stenosis of previously successfully dilated coronary arteries still remains a matter of concern despite of the improved short and long term results of percutaneous coronary angioplasty. The role of dyslipidaemia in the origin of restenosis after coronary angioplasty is still controversial. The aim of our study was to evaluate the efficacy of hypolipemic treatment in patients undergoing coronary angioplasty and to find out whether successful lowering of lipid parameters to normal limits is related to improvement exercise capacity and systolic function of left ventricle. The study group comprised 152 patients (17 women, 135 men), aged 52 +/- 8.8 years, who were reffered for percutaneous coronary angioplasty (PTCA). The patients were divided, according to the ratio of total cholesterol to HDL cholesterol (CH/ch-HDL), into two subgroups: subgroup I with CH/ch-HDL > 5.0 and subgroup II with CH/ch-HDL < or = 5.0. In all patients following parameters: total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides and body mass index were measured before PTCA, 1 month and 6 months after the procedure. At the same times a treadmill test and echocardiography were performed. Baseline total cholesterol, HDL cholesterol and triglycerides were significantly higher in subgroup I. In subgroup I Ch/ch-HDL ratio was at baseline 7.4 +/- 2.0 and decreased 6 months after PTCA to 5.2 +/- 1.7, p < 0.001. The CH/ch-HDL ratio was 4.2 +/- 0.6 in subgroup II before PTCA and remained the same 4.1 +/- 1.2 after 6 months. Before PTCA, the exercise capacity did not differ between groups 9.1 +/- 2.5 vs 9.6 +/- 3.3 MET, p = ns. A significant improvement of exercise capacity was observed in subgroup I 1 month after PTCA 9.1 +/- 2.5 to 11.2 +/- 2.7 MET, p < 0.001, remaining at the same level after 6 months. The differences seen in group II did not reach the statistical significance. Echocardiography revealed improvement of left ventricle contractility in both subgroups, with statistically significant increase in group I (1.24 +/- 0.36 to 1.14 +/- 0.27, p < 0.001). The left ventricle systolic function was within normal limits in all patients before coronary angioplasty and increased significantly 1 month after PTCA in subgroup I (56.1 to 60.4%, p < 0.001). 6 months after PTCA no further significant changes were observed. CONCLUSIONS: The dyslipidaemic state recognised before PTCA does not influence clinical outcomes after the procedure. Lipid lowering therapy should be offered to every patient undergoing coronary angioplasty regardless of the baseline value of total cholesterol to HDL cholesterol ratio. Optimal treatment of dyslipidaemia leading to lower total cholesterol, triglycerides and total cholesterol to HDL cholesterol ratio, to normal limits, is associated with improved exercise capacity and systolic function of left ventricle six months after successful coronary angioplasty. PMID- 11424663 TI - [Analysis of causes for anemia in patients with multiple myeloma]. AB - Pathogenetic mechanism of myeloma (MM) associated anaemia may involve: decreased level of erythropoietin (Epo) production, insufficient response to this hormone, bone marrow replacement by tumour cells, and cytokine mediated suppressive effect on erythropoiesis (ACD type of anaemia). We evaluated some possible causes of MM associated anaemia in a group of MM patients before treatment, using tests of in vitro culture of erythroid precursors and determination of the level of selected cytokines and evaluation of patient sera and media conditioned by patient cells for haematopoietic growth in vitro inhibitory/stimulatory activity. The study group included 15 patients with advanced MM at III clinical stage (according Durie and Salmon classification) prior treatment with a median age 61.8 women and 7 men. Diagnostic work-up included cytogenetics and serum Epo level as well as routine tests. We determined number of CFU-E, BFU-E and CFU-GM in the bone marrow of patients and healthy donors after mononuclear and T cell depletion in serum free culture under different conditions: 1) with conditioned medium (CM) by patients T cells, 2) CM by T cells of healthy donors and 3) without CM. IL-6, IL 1 alpha and TNF-alpha level in the serum and CM was determined. All patients were slightly anaemic--mean Hb 6 mmol/l (9.6 g/dl), transferrin level was increased- mean 1612 pmol/l (732 ng/ml) whereas iron level was normal. The average frequency of erythropoietic cells in myelogram was decreased (12%), however, the frequency of plasmocytes was increased (32%). Epo serum level was decreased in comparison with degree of anaemia (mean 22.2 U/ml). The average number of CFU-E/10(5) was slightly lower--212 and in the cultures containing CM by patient T cell and CM-by T cells of healthy donor decreased significantly to 138 and 183, accordingly. This phenomenon was not observed in cultures containing normal bone marrow cells. IL-6 level was increased in patients' serum and CM by patients T cells (27 ng/ml and 51 ng/ml, respectively). CONCLUSIONS: The results of this study support the notion on the multifactorial origin of anaemia in myeloma patients. The factors specified in this study include 1. defective response of Epo to the degree of anaemia 2. decreased number of committed erythroid cells in the marrow 3. suppression of erythropoiesis by IL-6. PMID- 11424664 TI - [Controlled release diltiazem in monotherapy of hypertension--time of drug administration and circadian blood pressure pattern]. AB - 30 patients with primary mild to moderate hypertension (DBP 95-110 mm Hg) were treated with long acting diltiazem (120 or 240 mg) once daily in the morning (7.00-8.00 a.m.) for at least 3 weeks and after that the administration time was changed to evening dose (19.00-20.00 p.m.) for next 3 weeks. 24-hours ABPM was performed in all patients on the last day of each period. Obtained recordings were compared in different periods of time: total 24 hours, 6.00-23.00, 23.00 6.00, 4.30-8.00, 5.00-11.00, 20.00-2.00. Mean values of systolic and diastolic blood pressure, heart rate and pressure load (defined as percentage of records above 140/90 mm Hg in day and above 120/80 mm Hg at night) did not differ significantly between investigated dosage regimens. The evening administration of diltiazem did not produce greater decrease of BP at night than the morning dose. For this reason slow release formulation of diltiazem (oxycardil in doses of 120 to 240 mg) can be safely administered in the evening if needed. PMID- 11424665 TI - [Advanced abdominal angina due to atherosclerosis with atypical celiac arteries]. AB - It is generally accepted that the abdominal angina develops only when at least two of the three splanchnic vessels-mesenteric arteries and the celiac trunk exhibit a critical obstruction. That common opinion does not, however, take into account anatomical variants of arteries supplying the blood to the intestines. We present a case of a wasted, 40 year old male with a wide spread arteriosclerosis and postprandial pain. The ultrasound examination revealed total occlusion of the superior mesenteric artery (SMA). Celiac trunk (CT) and inferior mesenteric artery (IMA) were patent. The ultrasound indicated that only one splanchic vessel was obstructed; the systemic disorder, the neoplasm, as well as the malabsorption were ruled out. An arteriography of the abdominal aorta and of splanchnic arteries confirmed patency of CT and IMA, also lack of flow in the SMA. Atypical origin of the middle colic artery originating from the bed of CT was also shown. Lack of collaterals between IMA and SMA, typically conducting a sufficient blood flow, resulted in a fully symptomatic abdominal angina. Symptoms were relieved following surgical revascularization. PMID- 11424666 TI - [Spontaneous pneumomediastinum with subcutaneous emphysema and pneumothorax as the initial manifestation of bronchial asthma. Case report]. AB - Spontaneous emphysema is not only the rate complication of bronchial asthma but may also be the first, atypical manifestation of the disease. We have presented here the case report of a young man in whom the occurrence of spontaneous pneumomediastinum with subcutaneous emphysema and pneumothorax led to the diagnosis of bronchial asthma. PMID- 11424667 TI - [Transport functions of the peritoneal membrane as a research topic in experimental medicine]. PMID- 11424668 TI - [Current pharmacotherapy of obesity]. PMID- 11424669 TI - [Goodpasture's syndrome]. PMID- 11424670 TI - Chemical, physical, and environmental properties of pelleted newspaper compared to wheat straw and wood shavings as bedding for horses. AB - Two experiments were conducted comparing pelleted recycled newspaper (PN) to wheat straw (S) and kiln-dried pine wood shavings (WS) as an animal bedding material. Adult horses housed 20 to 21 h/d in boxstalls served as the animal model for comparisons. In Exp. 1 eight boxstalls, each housing one horse, were each bedded with two types of PN (0.32 and 0.64 x 2.54 cm), S, and WS over four 5 d periods (replicated 4 x 4 Latin square). Initial amounts of bedding materials surpassed most commercial conditions, but stalls were cleaned daily of feces only and additional clean bedding was added as needed to maintain animal cleanliness, thus challenging the bedding properties of each material. In Exp. 2 nine boxstalls were bedded with PN (0.32 x 2.54 cm), S, and WS over three 14-d periods (three 3 x 3 Latin squares) during summer and autumn. Feces and wet spots were removed daily and clean bedding was added to reestablish working volume and simulate commercial conditions. In Exp. 1 and 2 daily additions of clean bedding varied (P < 0.05) with material (S > WS > PN). The higher water-holding capacity of PN and WS contributed to fewer bedding replacements. Usage of each material was greater (P < 0.05) during the autumn; PN had the greatest increase. Type of material and season also influenced bedding environment. Bedding pH increased (P < 0.05) with use and was highest in PN and lowest in S. Higher concentrations of breathable NH3 N were present in stalls bedded with PN and during autumn. Higher pH of used PN and decreased ventilation due to closed doors and windows during autumn were contributing factors. Season, type of bedding, and duration of its use affected (P < 0.05) numbers as well as species of microorganisms present in the breathing zone, nasal cavity, and on the leg of the horse. Clean and used WS contained greater (P < 0.05) quantities of particle fines, but with 5 d of use, particle fines in PN also increased. Quantities of breathable dust during cleaning of stalls varied (P < 0.05) with material and duration of its use; dust peaked at d 7 with PN but continued to decrease with S and to increase with WS through d 14. These data indicate that management of bedding materials varies with type of material and season of year. Use of PN as a bedding material has high potential. PMID- 11424671 TI - Genotype x environment interactions for postweaning performance in crossbred calves grazing winter wheat pasture or dormant native prairie. AB - Data from 403 calves from Angus, Brahman, and reciprocal-cross cows sired by Polled Hereford bulls were used to evaluate the impact of postweaning backgrounding forages on postweaning BW, gains, and carcass traits. Calves were born (spring of 1991 through 1994) and reared on either endophyte-infected tall fescue or common bermudagrass pastures. After weaning, calves were transported 360 km to the Grazinglands Research Laboratory, west of El Reno, OK, and, within breed and preweaning forage, were assigned to one of the following winter stocker treatments: 1) winter wheat pasture or 2) dormant native prairie plus supplemental CP. In March, winter stocker treatments were ended and calves were grazed as a single group on cool-season grasses until early July (1992, 1993, and 1994) or late May (1995), when the feedlot phase began. In the feedlot, calves were fed a high concentrate diet for an average of 120 d until a backfat thickness of > 10 mm was reached. Calves were shipped in truck load lots to Amarillo, TX (350 km), for processing and collection of carcass data. Averaged over calf breed group, calves wintered on wheat pasture gained faster (P < 0.01) during the stocker phase (0.71 vs 0.43 kg); had heavier (P < 0.01) final feedlot weights (535 vs 512 kg); lower feedlot (P < 0.01) ADG (1.37 vs 1.53 kg); heavier (P < 0.01) carcass weights (337 vs 315 kg); larger (P < 0.01) longissimus muscle (84.9 vs 81.8 cm2); higher percentage (P < 0.01) of kidney, heart, and pelvic fat (2.32 vs 2.26); and higher (P < 0.01) dressing percentage (62.2 vs 61.3) than calves backgrounded on native prairie. Maternal heterosis for stocker ADG was evident in calves backgrounded on native prairie but not on winter wheat (P < 0.10), but the two environments were similar in maternal heterosis for feedlot ADG and carcass traits. Calves wintered on native prairie were restricted in growth and expressed compensatory gain during the feedlot phase but not during the spring stocker phase. Dormant native grasses can be used to winter stocker calves excess to the winter wheat pasture needs, but ownership of these calves would have to be retained through the feedlot phase to realize any advantage of built-in compensatory gain. Finally, these data suggest that expression of maternal heterosis for weight gain is more likely in calves backgrounded on native prairie than in calves grazed on winter wheat. PMID- 11424672 TI - Gluteus medius and rump fat depths as additional live animal ultrasound measurements for predicting retail product and trimmable fat in beef carcasses. AB - This study was conducted to determine the ability of additional ultrasound measures to enhance the prediction accuracy of retail product and trimmable fat yields based on weight and percentage. Thirty-two Hereford-sired steers were ultrasonically measured for 12th-rib fat thickness, longissimus muscle area, rump fat thickness, and gluteus medius depth immediately before slaughter. Chilled carcasses were evaluated for USDA yield grade factors and then fabricated into closely trimmed, boneless subprimals with 0.32 cm s.c. fat. The kilogram weight of end-point product included the weight of trimmed, boneless subprimals plus lean trim weights, chemically adjusted to 20% fat, whereas the fat included the weight of trimmed fat plus the weight of fat in the lean trim. Prediction equations for carcass yield end points were developed using live animal or carcass measurements, and live animal equations were developed including ultrasound ribeye area or using only linear measurements. Multiple regression equations, with and without ultrasound rump fat thickness and gluteus medius depth, had similar R2 values when predicting kilograms of product and percentages of product, suggesting that these alternative variables explained little additional variation. Final unshrunk weight and ultrasound 12th-rib fat thickness explained most of the variation when predicting kilograms of fat. Rump fat and gluteus medius depth accounted for an additional 10% of the variation in kilograms of fat, compared with the equation containing final weight, ultrasound ribeye area, and ultrasound 12th-rib fat thickness; however, the two equations were not significantly different. Prediction equations for the cutability end points had similar R2 values whether live animal ultrasound measurements or actual carcass measurements were used. However, when ultrasound ribeye area was excluded from live animal predictions, lower R2 values were obtained for kilograms of product (0.81 vs 0.67) and percentages of product (0.41 vs 0.17). Conversely, the exclusion of ultrasound ribeye area had little effect on the prediction accuracy for kilograms of fat (0.75 vs 0.74) and percentage fat (0.50 vs 0.40). These data substantiate the ability of live animal ultrasound measures to accurately assess beef carcass composition and suggest that the alternative ultrasound measures, rump fat and gluteus medius depth, improve the accuracy of predicting fat-based carcass yields. PMID- 11424673 TI - Milk production of crossbred daughters of high- and low-milk EPD Angus and Hereford bulls. AB - Milk yield from 273 Angus- and Hereford-sired cows and preweaning performance of their calves were used to determine how accurately milk EPD of Angus and Hereford sires predicted milk production of crossbred daughters and subsequent calf performance. Mean milk EPD (kg) for high Angus (HA), low Angus (LA), high Hereford (HH), and low Hereford (LH) bulls (n = 41) selected as sires were +8.7, 6.2, +7.6, and -4.8, respectively. Cows calved in spring or fall from 1992 to 1997 and yielded a total of 660 records. Twenty-four-hour milk production of the cows was estimated by two weigh-suckle-weigh measurements at monthly intervals. The statistical model included breed, milk EPD level, sire of cow within breed and milk EPD level, year, season, cow age, calf sire, sex, and all two- and three way interactions. Means were obtained for monthly milk production, total milk production, time and yield of peak production, monthly calf weights, monthly cow weights and body condition scores (1 through 9), and calf birth and weaning data. The least squares means for 24-h milk production (kg) of HA, LA, HH, and LH with P-values for high vs low, across breeds, were, respectively, as follows: mo 1: 6.9, 5.9, 7.1, and 5.7 (P < 0.01); mo 2: 7.2, 6.1, 6.9, and 5.7 (P < 0.01); mo 3: 6.1, 5.1, 5.1, and 4.3 (P = 0.01); mo 4: 6.1, 4.9, 4.9, and 4.8 (P = 0.01); mo 5: 4.8, 4.0, 4.2, and 3.8 (P = 0.01); mo 6: 4.7, 3.4, 3.2, and 3.0 (P < 0.01); and mo 7: 3.7, 2.5, 3.0, and 3.0 (P = 0.05). Least squares means for total milk (kg) were 911.4, 729.6, 758.0, and 664.2 (P < 0.01); for yield at peak (kg/d) were 7.0, 5.7, 6.1, and 5.2 (P < 0.01); for birth weight (kg) were 37.1, 37.9, 38.3, and 38.8 (P = 0.31); for 205-d weight (kg) were 237.3, 218.2, 222.2, and 214.1 (P < 0.01); for final cow weight (kg) were 482.4, 505.4, 509.5, and 511.7 (P = 0.11); and for final cow BCS were 4.9, 5.3, 5.1, and 5.2 (P < 0.01). The correlations of total production with the monthly productions were 0.52, 0.56, 0.52, 0.54, 0.35, 0.37, and 0.31 (P < 0.01) and were 0.12 with birth weight, 0.45 with 205-d weight, -0.12 with final cow weight, and -0.26 with final cow body condition score (all P < 0.01). Daughters of high-milk EPD sires produced more milk and weaned heavier calves than those of low-milk EPD sires at the expense of body condition. These results suggest that sire milk EPD are sufficiently associated with milk yield and calf performance to be useful tools in genetic improvement of preweaning performance. PMID- 11424674 TI - Effects of programmed growth rate and days fed on performance and carcass characteristics of feedlot steers. AB - Two experiments were conducted to investigate a feeding regimen in which a programmed amount of feed was offered daily to control growth rate of steers. In Exp. 1, steers (n = 107, 309 +/- 3 kg) were used to determine effects of offering ad libitum access to feed (AL) vs a programmed intake feeding regimen (PI) and the number of days steers were fed (168 vs 203) on performance and carcass characteristics. Steers in the programmed intake feeding regimen were fed to achieve a predicted gain of 1.13 kg/d for the first 78 kg of gain, 1.36 kg/d for the next 124 kg of gain, and were given ad libitum access to feed for the final 54 or 103 kg of gain before slaughter (for steers fed for 168 d or 203 d, respectively). Feed efficiency was greater (P < 0.02) for steers in the PI than for those in the AL feeding regimen (0.193 vs 0.183 kg gain/kg feed, respectively). From d 169 to 203, steers in the PI feeding regimen had greater (P < 0.06) ADG (1.60 vs 1.38 kg/d) and similar (P = 0.38) feed efficiency than steers in the AL regimen. In Exp. 2, steers (n = 96; 308 +/- 3 kg BW) were offered feed ad libitum throughout the experiment (AL) or were programmed to gain at a high (PI-H) or low (PI-L) growth rate. For the first 78 kg of gain, intake was restricted to achieve predicted gains of 1.13 kg/d (PI-L) or 1.25 kg/d (PI H). For the next 124 kg of gain, intake was restricted to achieve predicted gains of 1.36 kg/d (PI-L) or 1.47 kg/d (PI-H). Feed was offered ad libitum for the final 58 kg of gain. Overall ADG was similar (P > 0.37) among feeding regimens despite lower DMI for the steers in the PI-L and PI-H feeding regimens than for those in the AL regimen. Feeding regimen did not affect (P < 0.22) carcass characteristics. Programmed intake feeding regimens sustained growth rate and feed efficiency for an extended period of time without detrimental effects on carcass characteristics. PMID- 11424675 TI - Proportion of corn silage in diets of feedlot steers fed to achieve stepwise increases in growth. AB - Two experiments were conducted to investigate the effects of proportion of dietary corn silage during periods of feed restriction on performance of steers. In Exp. 1, Simmental x Angus steer calves (n = 107; initial BW = 273 +/- 3.8 kg) were allotted to 12 pens with eight or nine steers/pen and four pens/treatment. Periods of growth were 273 to 366 kg BW (Period 1), 367 to 501 kg BW (Period 2), and 502 to 564 kg BW (Period 3). In two of the dietary regimens, steers were given ad libitum access to feed throughout the experiment and were fed either a 15% corn silage diet in each period or an 85, 50, and 15% corn silage diet in Periods 1, 2, and 3; respectively. In the third feeding regimen, a programmed intake feeding regimen was used. Steers were fed a 15% corn silage diet in each period. However, feed intake was restricted to achieve a predicted gain of 1.13 kg/d in Period 1 and 1.36 kg/d in Period 2, and feed was offered for ad libitum consumption in Period 3. For the entire experiment, ADG was similar (P = 0.41) among treatments and feed efficiency was lower (P < 0.10) for steers in the corn silage regimen than for steers in the programmed intake and ad libitum regimens. In Exp. 2, Simmental x Angus steer calves (n = 106; initial BW = 233 +/- 2 kg) were allotted by BW to 12 pens (three pens/treatment) and fed in three periods similar to those described in Exp. 1. Four feeding regimens were investigated: 1) AL; steers were offered a 15% corn silage diet for ad libitum consumption in all three periods; 2) PI; DMI was programmed to achieve gains as described in Exp. 1; 3) CS-HLL; programmed intake as described above except diets contained 85, 15, and 15% corn silage in Periods 1, 2, and 3, respectively; and 4) CS-HIL; same feeding regimens as CS-HLL, except diets contained 85, 50, and 15% corn silage in Periods 1, 2, and 3, respectively. Steers were given ad libitum access to feed in Period 3. Overall ADG was lower (P < 0.05) for steers in the CS-HLL and CS-HIL feeding regimens than for steers in the AL and PI regimens; feed efficiency was greatest for steers in the PI regimen. Few effects of feeding regimen on carcass characteristics were observed. PMID- 11424676 TI - Influence of clostridial vaccines and injection sites on performance, feeding behavior, and lesion size scores of beef steers. AB - Several clostridial vaccines are currently being used in the beef cattle industry. Of greatest concern is altering the location and route of administration of these vaccines to reduce injection-site lesions while maintaining seroconversion. Two experiments were conducted to determine the effect of clostridial vaccines and injection sites on the performance, feeding behavior, and lesion size scores of beef steers. In Exp. 1, 80 crossbred beef steers (BW 237 +/- 3.2 kg) were allotted randomly into five groups and given 14 d to adapt to the feed and individual feed intake-monitoring devices (Pinpointer devices) before starting the study. Each group was assigned randomly to one of the following vaccination treatments: 1) control (sterile saline water), 2) Alpha 7 Ear (A7E), 3) Alpha-7 Prescapula (A7P), 4) Vision-7 Prescapula (V7P), and 5) Ultrabac-7 Prescapula (U7P). All vaccines were injected s.c. in the ear or prescapular region, and injection sites were palpated on d 0 and 28 (Exp. 1) and on d 63 and 91 (Exp. 2). The protocol for Exp. 2 was exactly the same as for Exp. 1 except treatments included control, A7P, Alpha-CD Ear (ACDE), Alpha-CD Prescapula (ACDP), Fortress-7 Prescapula (F7P), and V7P. Also, control and steers receiving F7P and V7P were revaccinated on d 63 and palpated on d 91. Results of Exp. 1 indicated that the A7E and U7P steers had a feed intake lower (P < 0.01) than all other treatment groups. The ADG of the A7P and A7E steers were not different (P > 0.05) from those of the control steers. The gain:feed ratio of the A7E steers was 41% higher (P < 0.01) than that of the V7P steers (Exp. 1). The results of Exp. 2 indicated that the control, ACDP, and V7P steers had greater (P < 0.01) ADG than all other treatment groups, but the gain:feed ratios were not different (P > 0.05) among all treatment groups. Lesion sizes differed by vaccine and injection site in both experiments. These data suggest that vaccinating beef steers s.c. in the ear produced gain:feed ratios and lesion size scores that were similar to prescapular vaccinations. However, more research is required to determine the immune response of vaccinating cattle in the ear. PMID- 11424677 TI - Progress of farrowing and early postnatal pig behavior in relation to genetic merit for pig survival. AB - The objective of this study was to investigate whether pigs with different genetic merit for survival differed in birth weight, progress of farrowing, early postnatal behavior, or rectal temperature within 24 h after birth. On a nucleus farm in Rio Verde, Brazil, information was collected on 280 pigs, originating from 25 litters with known estimated breeding values for pig survival (EBVps). Litters were selected in such a way that a continuous range of EBVps with a maximum genetic contrast was achieved. Birth weight was recorded for all pigs. Indicators for progress of farrowing were birth intervals and duration of farrowing. Behavioral indicators of pig vitality were time until first upright standing (FUS), time until first udder contact (FUC), time until first teat in mouth (FTM), and time until first colostrum uptake (FCU). Rectal temperature was measured within 24 h after birth. Farrowing survival and early postnatal survival (within 3 d after farrowing) were registered. Farrowing survival and early postnatal survival both increased with increasing EBVps (farrowing survival: P = 0.007; early postnatal survival: P = 0.027). Birth weight decreased with increasing EBVps (P = 0.01). Birth intervals tended to increase with increasing EBVps (P = 0.10) and duration of farrowing was not related to EBVps. Time until first teat in mouth increased with increasing EBVps (P = 0.05), but the other behavioral indicators of pig vitality were not related to EBVps. Rectal temperature within 24 h after birth was not related to EBVps. Pigs with a higher genetic merit for survival have a lower birth weight but nevertheless have an increased farrowing survival and early postnatal survival. Their increased survival cannot be explained by differences in progress of farrowing, early postnatal behavior, or rectal temperature within 24 h after birth. PMID- 11424678 TI - Growth and slaughter traits of Boer x Spanish, Boer x Angora, and Spanish goats consuming a concentrate-based diet. AB - The number of Boer crossbred meat goats has been increasing rapidly, although how their growth and slaughter traits compare with those of Spanish goats and influences of maternal genotype have not been thoroughly evaluated. This information would be useful to achieve optimal meat goat production systems and yield of goat products desired by consumers. Therefore, postweaning growth (9 to 24 wk of age) and slaughter traits (212 +/- 5.0 d of age) of Boer x Spanish, Spanish, and Boer x Angora wethers (n = 16, 18, and 18 for growth measures, respectively, and n = 6 per genotype for slaughter traits) consuming a concentrate-based diet were compared. Over the 16-wk performance period, ADG, DMI, and ADG:DMI were greater (P < 0.05) for Boer crossbreds than for Spanish goats (ADG: 154, 117, and 161 g; DMI: 646, 522, and 683 g/d; ADG:DMI: 263, 235, and 261 g/kg for Boer x Spanish, Spanish, and Boer x Angora, respectively). Dressing percentage (46.3, 47.3, and 47.0% of BW; SE = 1.21) and quality grade score (11.17, 9.67, and 11.17 for Boer x Spanish, Spanish, and Boer x Angora, respectively; SE = 0.66 [12 = Choice+; 11 = Choice; 10 = Choice-; 9 = Good+]) were similar among genotypes. Weights of some noncarcass components were greater for Boer crossbreds than for Spanish goats, but relative to empty BW, noncarcass component weights were similar among genotypes. Concentrations of moisture, ash, fat, and protein in carcass and noncarcass components did not differ among genotypes. Contributions to the carcass of different primal cuts were similar among genotypes, and there were few differences in concentrations of separated lean, bone, and fat in primal cuts. In conclusion, when consuming a concentrate based diet, early postweaning growth rate was similar between Boer x Spanish and Boer x Angora wethers and greater for Boer crossbreds than for Spanish wethers. Slaughter traits were primarily related to differences in final BW. PMID- 11424679 TI - Comparison of F1 Bos indicus x Hereford cows in central Texas: I. Reproductive, maternal, and size traits. AB - Cows (n = 116) sired by Angus, Gray Brahman, Gir, Indu-Brazil, Nellore, and Red Brahman bulls and from Hereford dams were evaluated (when bred to the same breed of bull) for reproductive performance and cow weight and height and their calves (n = 1,161) were evaluated for birth and weaning weight in central Texas (temperate winters and subtropical summers). Cows were born from 1982 to 1985 and their calves were born from 1985 to 1999. Crossbred cows sired by Angus had a higher (P < 0.10) occurrence of calving difficulty than Gir, Indu-Brazil, Nellore, and Red Brahman crossbreds. Calves from Gir crossbreds had lighter (P < 0.10) birth weight (34.8 kg) than calves from Angus and Red Brahman crossbreds (39.4 and 37.2 kg). Calves from Angus crossbreds cows had lighter (P < 0.001) weaning weight (227.0 kg) than calves from Bos indicus crossbreds. Cows sired by Angus were lighter (P < 0.10) as mature cows (520.69 kg) than Gray Brahman, Indu Brazil, and Red Brahman crossbreds (585.6, 571.9, and 577.6 kg, respectively). They also had smaller (P < 0.05) hip height (124.7 cm) than Bos indicus crossbreds. Mature cows sired by Indu-Brazil had greater (P < 0.05) hip height (138.3 cm) than Gir crossbreds (133.8 cm). Differences among sire breeds were noted within cow age categories (2 through 14 yr of age) for pregnancy rate, calf crop born, calf survival rate (as a trait of cow), and calf crop weaned. Angus and Indu-Brazil crossbred cows were lower (P < or = 0.10) than Gir and Nellore crossbreds for pregnancy rate, calf crop born, and(or) calf crop weaned at some, but not all, ages. Indu-Brazil crossbreds had a lower (P < or = 0.10) calf survival rate than most other breed groups at ages 11 to 14 yr. The results of this study indicate that Nellore and Gir crossbreds, but not Indu-Brazil, could perform as well or better than breeds traditionally used in cross-breeding programs of U.S. cow-calf production. PMID- 11424680 TI - Comparison of F1 Bos indicus x Hereford cows in central Texas: II. Udder, mouth, longevity, and lifetime productivity. AB - Postpartum udder characteristics, aged mouth condition scores, and cow survival, longevity, and productivity were evaluated for 15 yr on F1 cows in central Texas (temperate winters and subtropical summers). The cows (n = 116) were progeny of Angus, Gray Brahman, Gir, Indu-Brazil, Nellore, and Red Brahman sires and Hereford dams. Crossbred cows sired by Nellore had smaller (P < 0.05) postpartum teat length than cows of all other crossbred groups and smaller (P < 0.10) postpartum teat diameter than Indu-Brazil and Red Brahman crossbreds. The Nellore crossbred cows had larger (P < 0.10) postpartum udder support scores (more well supported udders) than Gir, Indu-Brazil, and Red Brahman crossbreds. As 14-yr-old cows, Angus crossbreds had lower (P < 0.05) mouth scores (indicative of one or more missing incisors) than Bos indicus crossbreds. Under actual and artificial (records were removed) culling procedures, Nellore crossbreds had the highest cow survival to age 14 and the highest longevity. Indu-Brazil crossbreds tended to be the lowest for both survival and longevity. Curves were fitted by nonlinear regression procedures to the adjusted frequencies of cows remaining at each age; scale and shape parameters were estimated and compared for the different breed groups. Breed group cow survival rates, calf crop weaned, and calf weaning weights were used 1) to construct population projection and production matrices for each breed group and 2) to generate breed group vectors containing projected age distributions and production information. Cows sired by Nellore and Gir were projected to require the fewest purebred cows to generate replacement heifers and to have the most calves and the largest total calf weight sold per year. PMID- 11424681 TI - Estimated genetic parameters for growth traits of German shepherd dog and Labrador retriever dog guides. AB - A desirable dog guide weighs 18 to 32 kg and measures 53 to 64 cm in height at the withers as an adult. Heritabilities and genetic correlations were estimated for birth weight, 42-d weight, mature weight, and mature height for 2,334 German shepherd dogs and 2,028 Labrador retrievers raised by the Seeing Eye, Inc., Morristown, NJ, from 1979 to 1997. Data included 5,006 observations for German shepherd dogs from 113 dams and 33 sires and 4,123 observations for Labrador retrievers from 89 dams and 29 sires. A mixed effects model was considered with sex and birth year as fixed effects. Random effects were animal, maternal, and litter incorporating all pedigree information available. A derivative-free REML method was used to estimate parameters. The maternal component was higher than the additive genetic component for birth weight. Heritability of mature weight was estimated as 0.57 +/- 0.07 for German shepherd dogs and 0.44 +/- 0.07 for Labrador retrievers. Mature height heritability was estimated as 0.35 +/- 0.08 for German shepherd dogs and 0.46 +/- 0.08 for Labrador retrievers. Selection for lighter dogs at maturity is predicted to decrease the average mature height, and selection for taller dogs at maturity is predicted to increase the average mature weight. The estimated genetic parameters will aid in the development of strategies to increase the probability of breeding dogs for optimum mature size. PMID- 11424682 TI - Behavioral, adrenal, immune, and productive responses of lactating ewes to regrouping and relocation. AB - The experiment involved 40 mid-lactating Comisana ewes. The animals were housed in straw-bedded pens and assigned to a control group (n = 10) and two test groups of 15 subjects each. Control ewes were never moved from their pen and peers throughout the experiment period, whereas test animals were subjected to either regrouping (RG) or regrouping and relocation (RGRL) three times at weekly intervals (d 7, 14, and 21 of the trial). Three behavioral recordings were conducted after each social and pen exchange over a 6-h period (1000 to 1600). Animals from groups RG and RGRL were bled immediately before each regrouping and relocation and 15 and 60 min afterward. The phytohemagglutinin skin test was performed at d 9, 16, and 23. Milk yield was recorded when the ewes were mixed and moved, and before and after each mixing and moving procedure. Individual milk samples were analyzed for composition, renneting parameters, and bacteriological characteristics; samples with more than 10(6) somatic cells/mL were cultured for mastitis-related pathogens. Control ewes spent more time lying than groups RG (P < 0.001) and RGRL (P < 0.01) and less time in ambulatory activities than RGRL sheep (P < 0.05). Moved and remaining ewes showed a higher number of aggressive interactions than control subjects (P < 0.001 and P < 0.01, respectively). Control sheep showed the highest cell-mediated immune response compared with both moved and mixed ewes (P < 0.001 and P < 0.01, respectively). The RGRL ewes gave lower yields of milk than did the control ewes on the days after the first (P < 0.05) and the second (P < 0.01) change of pen and peers. The RGRL ewes also had a lower fat content (P < 0.05) than control ewes in the milk yielded on the day of the first regrouping and relocation and a lower milk protein content (P < 0.05) on the following day. The RG treatment resulted in the ewes yielding milk with a lower fat content (P < 0.01) on the day of each mixing procedure as compared with controls and in a decreased milk protein content on the days after the first (P < 0.05) and the second regrouping (P < 0.01). One case of subclinical mastitis was recorded in both the RG and RGRL groups, whereas no cases were detected in control group. These findings suggest that regrouping and relocation may cause increased aggression, altered immune responses, and short-term effects on the production performance of lactating ewes. These practices should be performed by attempting to minimize their impact on animal welfare. PMID- 11424683 TI - Behavior, reproduction, and immunity of crated pregnant gilts: effects of high dietary fiber and rearing environment. AB - The objective of this study was to examine effects of increased gut fill and diverse developing environments on pregnant gilts' behavior and physiology. Gilts were cross-fostered at 1 d of age and transferred to either an indoor or outdoor production unit. Littermate gilts remained in their different environments during development and were moved into individual gestation crates in an indoor gestation unit. Of the 42 gilts, 19 were fed a control diet of fortified sorghum soybean meal and 23 were fed the same diet with 25% beet pulp (high fiber). Control sows ate 2.0 kg/d and high-fiber sows ate 2.67 kg/d in a large pellet (thus resulting in approximately equal energy intake and differing total dietary intakes). Pregnant gilts had behavior and immune measures sampled at 30, 60, and 90 d of gestation. The day x diet interaction was significant (P = 0.01) for duration of standing: sows fed high-fiber diets stood less on d 30, but on d 60 and 90 they and the control sows stood for a similar duration. Sham chewing duration and frequency showed significant (P < 0.05) effects of gestation stage x diet x environment. Gilts reared outdoors and fed high fiber increased sham chewing over gestation, whereas all other treatment groups decreased this behavior over time. Outdoor-reared gilts had greater (P < 0.05) frequency and duration of drinking behavior than indoor-reared gilts. White blood cell numbers were higher (P < 0.05) for gilts fed high-fiber diets than for gilts fed the control diet. Immune (humoral and cellular systems) and reproductive measures (farrowing rate and litter size) and plasma cortisol concentrations were generally not influenced (P > 0.10) by diets and rearing environments, suggesting that in spite of significant changes in behavior and feed intake gilts' immune systems were not suppressed or enhanced. Behavioral data alone suggested that indoor-reared gilts showed fewer behavioral adaptations to the crates than outdoor-reared gilts. However, immune measures did not indicate that any treatments resulted in physiological effects indicative of stress. PMID- 11424684 TI - Total body electrical conductivity (TOBEC) measurement of compositional differences in hams, loins, and bellies from conjugated linoleic acid (CLA)-fed stress-genotype pigs. AB - This study was designed to observe the effects of conjugated linoleic acid (CLA) supplementation on lean content of pork carcass primal cuts (hams, loins, and bellies) and to determine the ability of total body electrical conductivity (TOBEC) to predict lean content. A total of 64 crossbred growing-finishing barrows were placed on a control (soybean oil) or CLA (0.75%) diet at an average weight of 40 kg. Pigs were penned in pairs according to diet and stress genotype (negative, carrier, and positive) and slaughtered at 115 kg. Stress genotype was included because of known variations in lean content. Hams (IMPS 401A; n = 64), loins (IMPS 410; n = 24), and bellies (IMPS 408; n = 63) were fabricated from carcasses at 24 h postmortem and scanned for electromagnetic (EM) absorption by a MQ-25 EM scanner. Each wholesale cut was scanned in triplicate at 2.5 MHz to yield a peak mean average (PMA) value then separated into lean, fat, bone, and skin components. Bellies were skinned prior to scanning then subjected to a belly bar firmness test before dissection. Supplementation with CLA had no effect (P > 0.05) on lean ham composition. Regression analysis was used for lean weight prediction using primal weight and PMA value as predictors. Lean content prediction of hams by TOBEC resulted in an R2 of 0.80. Loins from CLA supplemented pigs exhibited increased lean weight (P < 0.05) and PMA values (P < 0.05) compared to controls. Lean prediction of loins by TOBEC resulted in an R2 of 0.66. Bellies from CLA-supplemented pigs had a higher percentage of moisture (P < 0.03) and protein (P < 0.01) and decreased percentage of lipid (P < 0.01). The R2 values from the regression analysis predicting protein, moisture, protein + moisture, and fat-free soft tissue composition of the skinless bellies were 0.67, 0.68, 0.71, and 0.78, respectively. PMID- 11424685 TI - Evaluation of ultrasound imagery and body shape to predict carcass and fillet yield in farm-raised catfish. AB - Accurate prediction of meat yield in live animals may allow more efficient genetic improvement of meat yield in farm-raised catfish. An initial trial with 30 channel catfish demonstrated significant correlations among weight-adjusted residuals for muscle area measured from transverse ultrasound images and transverse sections at five locations along the trunk musculature (r = 0.30 to 0.70). Relationships of weight-adjusted residuals for three meat yield traits (carcass, whole fillet, and shank fillet) with weight-adjusted residuals for 15 external body shape measurements and five transverse ultrasound measurements of muscle area were determined for 51 female and 91 male channel x blue catfish backcross hybrids. Compared to males, females had smaller heads; deeper, wider, shorter bodies; larger ultrasound muscle area; and higher meat yield. Correlations between carcass traits and body shape and carcass traits and ultrasound measurements were generally higher for females than for males. Correlations among carcass traits and ultrasound muscle area were typically higher than correlations among carcass traits and external body shape in both sexes. A single ultrasound measurement explained 40 to 50% and 16 to 23% of the variation in meat yield traits of females and males, respectively. The best three variable model using ultrasound and body shape traits explained 48 to 56% and 31 to 38% of the variation in meat yield traits in females and males, respectively. Differences between males and females for the variability in meat yield traits explained by the models may be related to sexual dimorphism for body shape and fillet yield observed in catfish. Ultrasound has potential for predicting meat yield in live fish, but improved prediction accuracy is needed. Differences in meat yield traits between males and females and among individuals within sexes suggest that selecting for fish with smaller heads and deeper, shorter body shape posterior to the visceral cavity will increase meat yield in catfish. PMID- 11424686 TI - Effects of choice white grease or poultry fat on growth performance, carcass leanness, and meat quality characteristics of growing-finishing pigs. AB - Eighty-four crossbred gilts were used to evaluate the effects of dietary choice white grease (CWG) or poultry fat (PF) on growth performance, carcass characteristics, and quality characteristics of longissimus muscle (LM), belly, and bacon of growing-finishing pigs. Pigs (initially 60 kg) were fed a control diet with no added fat or diets containing 2, 4, or 6% CWG or PF. Diets were fed from 60 to 110 kg and contained 2.26 g lysine/Mcal ME. Data were analyzed as a 2 x 3 factorial plus a control with main effects of fat source (CWG and PF) and fat level (2, 4, and 6%). Pigs fed the control diet, 2% fat, and 4% fat had greater (P < 0.05) ADFI than pigs fed 6% fat. Pigs fed 6% fat had greater (P < 0.05) gain/feed (G/F) than pigs fed the control diet or other fat levels. Subcutaneous fat over the longissimus muscle from pigs fed CWG had more (P < 0.05) moisture than that from pigs fed PF. Feeding dietary fat (regardless of source or level) reduced (P < 0.05) the amount of saturated fats present in the LM. Similarly, 4 or 6% fat decreased (P < 0.05) the amount of saturated fats and increased unsaturated fats present in the bacon. No differences (P > 0.05) were observed for ADG, dressing percentage, leaf fat weight, LM pH, backfat depth, LM area, percentage lean, LM visual evaluation, LM waterholding capacity, Warner-Bratzler shear and sensory evaluation of the LM and bacon, fat color and firmness measurements, or bacon processing characteristics. Adding dietary fat improved G/F and altered the fatty acid profiles of the LM and bacon, but differences in growth rate, carcass characteristics, and quality and sensory characteristics of the LM and bacon were minimal. Dietary additions of up to 6% CWG or PF can be made with little effect on quality of pork LM, belly, or bacon. PMID- 11424687 TI - Effect of postmortem storage on mu-calpain and m-calpain in ovine skeletal muscle. AB - Casein zymography was used to determine the effect of postmortem storage on the proteolytic activity of mu-calpain and m-calpain in lamb longissimus. Casein zymography assays were conducted on crude muscle extracts (only one centrifugation). Six market weight crossbred lambs were slaughtered and a portion of the longissimus lumborum was removed at death (within 15 min of exsanguination) and after 3, 6, 9, 12, 24, 72, and 360 h postmortem. Muscle samples were snap-frozen in liquid nitrogen and stored at -70 degrees C. Soluble muscle proteins were extracted from muscle samples and analyzed by in-gel casein assay to measure calpain proteolytic activity. There was a gradual decline in mu calpain activity (P < 0.05) such that after 24 and 72 h postmortem, mu-calpain had lost 42 and 95% of its activity, respectively. After 360 h postmortem, no mu calpain activity could be detected (under the conditions used in this study). Autolysis of mu-calpain could be detected as early as 3 h postmortem. It was demonstrated that the detectable level of mu-calpain activity is a function of the amount of muscle protein electrophoresed. Hence, the activity data reported are in relative terms, rather than absolute values. Furthermore, it was demonstrated that the activity data also are a function of the assay methods used. Different methods have different lower detection limits. Of the three assays examined, 14C-labeled casein was the most sensitive, then the in-gel casein assay, and the least-sensitive method was the standard casein assay. Unlike mu-calpain, postmortem storage had no effect on m-calpain (P > 0.05). When the calcium concentration of a muscle extract was increased to the level that induces m-calpain autolysis, m-calpain was autolyzed and its autolysis was readily detected by the in-gel casein assay. Collectively, these results demonstrate that calcium concentration in postmortem muscle is only high enough to activate mu-calpain. These results support the widely believed conclusion that mu-calpain-mediated proteolysis of key myofibrillar and cytoskeletal proteins is responsible for postmortem tenderization. Hence, understanding the regulation of mu-calpain in postmortem muscle should be the focus of future studies. PMID- 11424688 TI - Effect of dietary L-carnitine on growth performance and body composition in nursery and growing-finishing pigs. AB - Two experiments were conducted to determine the effect of dietary L-carnitine on growth performance and carcass composition of nursery and growing-finishing pigs. In Exp. 1,216 weanling pigs (initially 4.9 kg and 19 to 23 d of age) were used in a 35-d growth trial. Pigs were blocked by weight in a randomized complete block design (six pigs per pen and six pens per treatment). Four barrows and four gilts were used to determine initial carcass composition. L-Carnitine replaced ground corn in the control diets to provide 250, 500, 750, 1,000, or 1,250 ppm. On d 35, three barrows and three gilts per treatment (one pig/block) were killed to provide carcass compositions. L-Carnitine had no effect (P > 0.10) on growth, percentages of carcass CP and lipid, or daily protein accretion. However, daily lipid accretion tended to decrease and then return to values similar to those for control pigs (quadratic P < 0.10) with increasing dietary L-carnitine. In Exp. 2, 96 crossbred pigs (initially 34.0 kg BW) were used to investigate the effect of increasing dietary L-carnitine in growing-finishing pigs. Pigs (48 barrows and 48 gilts) were blocked by weight and sex in a randomized complete block design (two pigs/pen and eight pens/treatment). Dietary L-carnitine replaced cornstarch in the control diet to provide 25, 50, 75, 100, and 125 ppm in grower (34 to 56.7 kg; 1.0% lysine) and finisher (56.7 to 103 kg; 0.80% lysine) diets. At 103 kg, one pig/pen was slaughtered, and standard carcass measurements were obtained. Dietary L-carnitine did not influence growth performance (P > 0.10). However, increasing dietary carnitine decreased average and tenth-rib back-fat (quadratic, P < 0.10 and 0.05), and increased percentage lean and daily CP accretion rate (quadratic, P < 0.05). Break point analysis projected the optimal dosage to be between 49 and 64 ppm of L-carnitine for these carcass traits. It is concluded that dietary carnitine fed during the nursery or growing-finishing phase had no effect on growth performance; however, feeding 49 to 64 ppm of L-carnitine during the growing-finishing phase increased CP accretion and decreased tenth-rib backfat. PMID- 11424689 TI - Alkanes as internal markers to estimate digestibility of hay or hay plus concentrate diets in horses. AB - Dry matter intake (DMI), dry matter digestibility (DMD), and fecal output (FO) are difficult to measure directly in the field, and indirect methods using external and internal markers have thus been developed. An experiment was conducted consisting of two digestion trials with two periods in each trial to examine the use of five odd-chain alkanes (C25 to C33) of plant cuticular wax as internal markers to estimate DMD of hay or hay plus concentrate diets in horses. Eight mature Thoroughbred geldings were housed in 4- x 4-m stalls and randomly assigned to one of two mixed grass/legume hays (Diets 1 and 2) in Trial 1 and to mixed grass/legume hay plus one of two concentrates (Diets 3 and 4) in Trial 2. After the first 12-d period was conducted, dietary assignments for each group were switched for the second period in each trial. Each period consisted of a dietary accommodation from d 1 to 7 and total fecal collection from d 8 to 11. Results indicated that fecal recoveries of odd-chain alkanes were 88 to 90% for Diet 1, 75 to 92% for Diet 2, 71 to 81% for Diet 3, and 71 to 82% for Diet 4. Alkane recoveries were not related to alkane chain lengths. Digestibilities calculated from alkane concentration data adjusted using the mean fecal recovery of individual odd-chain alkanes (DA1) were not significantly different from the digestibilities estimated from total collection (DTC) for Diets 1 and 2 in Trial 1 and Diets 3 and 4 in Trial 2. When adjustment was based on the mean recovery of all alkanes (DA2; estimated by linear regression), all DA2 estimates for horses offered all diets were similar to DTC. Results indicate that accurate mean estimates of DMD can be obtained by using plant wax alkane markers and adjusting for the mean recovery of five odd-chain alkanes in a diet. PMID- 11424690 TI - Ileal and total tract nutrient digestibilities and fecal characteristics of dogs as affected by soybean protein inclusion in dry, extruded diets. AB - Plant-based protein sources are generally less variable in chemical composition than animal-based protein sources. However, relatively few data are available on the nutrient digestibilities of plant-based protein sources by companion animals. The effects of including selected soybean protein sources in dog diets on nutrient digestion at the ileum and in the total tract, as well as on fecal characteristics, were evaluated. Six protein sources were used: soybean meal (SBM), Soyafluff 200W (soy flour), Profine F (traditional aqueous-alcohol extracted soy protein concentrate [SPC 1]), Profine E (extruded SPC [SPC 2]), Soyarich I (modified molecular weight SPC [SPC 3]), and poultry meal (PM). All diets were extruded and kibbled. Test ingredients varied in CP and fat contents; however, diets were formulated to be isonitrogenous and isocaloric. Nutrient intakes were similar, except for total dietary fiber (TDF), which was lower (P < 0.01) for dogs fed the PM diet. Apparent ileal digestibilities of DM, OM, fat, and TDF were not different among treatments; however, CP digestibility at the terminal ileum was higher (P < 0.01) for diets containing soy protein sources than for PM. Total tract CP digestibility was greater (P < 0.01) for soy protein containing diets than for PM. Apparent total tract digestibilities of DM, OM, fat, and TDF were not different among treatments. Apparent amino acid digestibilities at the terminal ileum, excluding methionine, threonine, alanine, and glycine, were higher (P < 0.01) for soy protein-containing diets than for PM. Dogs fed SPC diets had lower (P < 0.01) fecal outputs (g asis feces/g DMI) than dogs fed the SF diet, and dogs fed SBM tended (P < 0.11) to have lower fecal outputs than dogs fed the SF diet. However, dogs fed the PM diet had lower (P < 0.03) fecal outputs than dogs fed SPC-containing diets. Fecal outputs and scores reflected the TDF and nonstructural carbohydrate contents of the soy protein fraction. Soy protein sources are well utilized by the dog prior to the terminal ileum, and SPC offers a viable alternative to PM as a protein source in dry, extruded canine diets. PMID- 11424691 TI - Reproductive efficiency and maternal-offspring transfer of gossypol in rainbow trout (Oncorhynchus mykiss) fed diets containing cottonseed meal. AB - In a preceding study, complete substitution of fish meal protein with cottonseed meal (CM) protein did not affect the survival or growth rate of adult rainbow trout over a 6-mo period. Gossypol, a naturally occurring compound in cottonseeds, has an antifertility effect in terrestrial animals, but information regarding salmonid fish is lacking. Female rainbow trout in this experiment were fed diets with either 0, 25, 50, 75, or 100% (diets 1 to 5) of the fish meal protein replaced with CM protein until first maturation and spawning to study long-term effects on growth and reproduction. Feeding diets containing CM over a total period of 10 mo did not result in differences in growth and mortality compared to the control group (P > 0.05). Increased CM incorporation levels resulted in decreased (P < 0.05) blood hemoglobin (10.6 +/- 1.3, 8.4 +/- 1.8, 7.3 +/- 1.1, 6.9 +/- 0.8, and 5.6 +/- 1.4 g/dL) and hematocrit (49.6 +/- 3.9, 38.5 +/ 9.3, 34.4 +/- 3.7, 34.8 +/- 4.9 and 28.0 +/- 6.8%) levels in diets 1 to 5, respectively. The CM incorporation level had no effect (P > 0.05) on the number of eggs produced per female but led to a reduction (P < 0.05) in egg weight. Eyed stage survival of embryos was low in all dietary groups and did not show differences (P > 0.05). However, an increasing CM incorporation level led to a linear increase (P < 0.05) in the number of females that produced no viable embryos (23.1, 37.5, 42.9, 60.0, and 71.4%). Gossypol in the diet was absorbed by the female trout and transferred to the eggs (0, 2.2 +/- 0.5, 6.7 +/- 1.6, 10.6 +/- 4.2, and 20.0 +/- 2.6 micrograms/g in diets 1 to 5, respectively). A high concentration of gossypol remained in the juveniles at the swim-up stage (endogenous yolk-absorbed) (0.6 +/- 0.3, 2.4 +/- 0.3, 3.4 +/- 0.0, and 4.7 +/- 1.0 micrograms/g, diets 2 to 5, respectively). The findings suggest that replacement of the dietary fish meal protein with CM protein has no effect on fish growth and mortality but may lead to a reduction in reproductive performance in female rainbow trout. PMID- 11424692 TI - Effects of exposure to high ambient temperature and dietary protein level on sow milk production and performance of piglets. AB - The effects of high ambient temperature and level of dietary heat increment on sow milk production and piglet performance over a 28-d lactation were determined in 59 multiparous crossbred Large White x Landrace pigs kept at a thermoneutral (20 degrees C) or in a hot (29 degrees C) constant ambient temperature. Experimental diets fed during lactation were a control diet (NP; 17.6% CP) and two low-protein diets obtained by reduction of CP level (LP; 14.2% CP) or both reduction of CP and addition of fat (LPF; 15.2% CP); the NE:ME ratio was 74.3, 75.6, and 75.8% for NP, LP, and LPF diets, respectively. All diets provided 0.82 g of digestible lysine/MJ of NE, and ratios between essential AA and lysine were above recommendations. Creep feed was provided after d 21 of lactation. Reduction of CP level did not influence (P > 0.10) milk production, milk composition, or piglet performance. Despite higher nursing frequency (39 vs 34 sucklings per day), milk production decreased (P < 0.01) from 10.43 to 7.35 kg/d when temperature increased from 20 to 29 degrees C. At d 14, DM (18.6 vs 18.1%) and energy (4.96 vs 4.75 MJ/kg) contents in milk tended (P = 0.09) to be higher in sows kept at 29 degrees C. Over the 28-d lactation, piglet BW gain and BW at weaning decreased (P < 0.01) from 272 to 203 g/d and 9.51 to 7.52 kg, respectively, when temperature increased from 20 to 29 degrees C. Daily creep feed intake over the 4th wk of lactation was higher (P < 0.01) at 29 degrees C than at 20 degrees C (388 vs 232 g/litter, respectively), which was reflected in a greater increase in BW gain between wk 1 to 3 and wk 4 at the higher temperature (147 vs 130%); BW gain between weaning and d 14 postweaning was higher (P < 0.05) for piglets originating from sows kept at 29 degrees C (280 vs 218 g/d). In connection with their lower growth rate, DM (31.2 vs 33.0%), protein (15.5 vs 16.0%), lipid (12.3 vs 13.9%), and energy (8.39 vs 9.09 kJ/g) contents in weaned, slaughtered piglets were lower (P < 0.01) at 29 than at 20 degrees C. In conclusion, modification in the CP:NE ratio in order to decrease dietary heat increment did not affect milk production and piglet performance in thermoneutral or hot climatic conditions. Our results confirm the negative effect of high ambient temperatures on milk yield and emphasize the importance of creep feed supply to improve pre- and postweaning growth of piglets in these conditions, especially when weaning occurs after 3 wk of age. PMID- 11424693 TI - Inflammatory response, growth, and thyroid hormone concentrations are affected by long-term boron supplementation in gilts. AB - An experiment was conducted to determine the long-term effects of dietary boron (B) on growth performance, immune function, and plasma and serum characteristics in gilts. Fifty weanling gilts were allotted to 10 pens based on weaning weight and litter origin. Pens were randomly assigned to receive one of two dietary treatments. Treatments consisted of a basal diet low in B (control) and the basal diet supplemented with 5 mg B/kg diet as sodium borate. Gilts remained on their respective experimental diets and with their penmates throughout the nursery, growing, and finishing phases. The B concentration of the basal diet was 0.98, 2.1, and 2.2 mg/kg diet during the nursery, growing, and finishing phases, respectively. At the end of each production phase, animals were weighed and feed consumption was determined to assess growth performance variables. In addition, blood samples were obtained from three randomly selected gilts per pen at the completion of each phase. Boron had no affect (P > 0.58) on growth performance during the nursery phase, but gilts receiving supplemental B had increased (P < 0.05) ADG at the end of the finishing phase and over the entire growing-finishing period. Serum concentrations of triiodothyronine (T3) tended (P < 0.07) to be reduced by dietary B at the end of the nursery phase, but serum thyroxine (T4) was not affected (P = 0.46) by B. At the completion of the growing phase, supplemental B decreased (P < 0.05) the concentrations of T3 and T4 in the serum. In addition, serum concentrations of total cholesterol and the activity of alkaline phosphatase were increased (P < 0.05) by dietary B at the end of the growing phase. Serum concentrations of urea N tended (P < 0.09) to be increased by B at the end of the growing phase. Beginning at d 95 of the experimental period, measures of immune function were assessed in randomly selected gilts. Boron decreased (P < 0.05) the inflammatory response to an intradermal injection of phytohemagglutinin. Boron did not affect (P > 0.30) the blastogenic response of isolated lymphocytes to mitogen stimulation or the humoral immune response against a sheep red blood cell suspension. Results indicate that B may affect serum thyroid hormone concentrations, the inflammatory response, and growth in pigs. PMID- 11424694 TI - Estimation of the total sulfur amino acid requirement and the effect of betaine in diets deficient in total sulfur amino acids for the weanling pig. AB - Four experiments were conducted to determine whether betaine (BET) could replace dietary methionine (MET) in diets for weanling pigs. Pigs in each experiment were allotted to treatments on the basis of weight in a randomized complete block design. Each treatment was replicated four (Exp. 4), five (Exp. 1 and 2), or six (Exp. 3) times with five or six pigs per replicate. In Exp. 1, pigs were fed a diet formulated to be deficient in total sulfur amino acids (TSAA) (negative control; NC) or the NC + 0.05 or 0.10% MET or BET during Phase 1 and 0.035 or 0.07% MET or BET during Phase 2. Growth performance was not affected (P > 0.10) by dietary treatments, indicating that the diets were not deficient in TSAA. In Exp. 2, graded levels of TSAA (0.74, 0.79, 0.84, 0.89, or 0.94%) were fed. Overall ADG was increased (0 vs added MET, P < 0.07) in pigs fed TSAA levels of 0.79% or greater, but gain:feed was not affected (P > 0.10) by diet. Overall ADFI was increased (linear, P < 0.08) and plasma urea N (PUN) was decreased (quadratic, P < 0.01) as the level of TSAA was increased. Most of the change in ADG, PUN, and ADFI occurred between 0.74 and 0.84% TSAA. Thus, the 0.74% TSAA diet was used in Exp. 3 as the NC. In Exp. 3, the diets included the following: 1) NC, 2) NC + 0.05% MET, 3) NC + 0.10% MET, 4) NC + 0.039% BET, or 5) NC + 0.078% BET. The addition of MET resulted in increased (linear, P < 0.10) ADG, ADFI, and gain:feed, but MET decreased PUN (linear, P < 0.05). Daily gain, ADFI, and TSAA intake were not different (P > 0.10) between pigs fed 0.05% MET or 0.039% BET, but gain:feed was decreased (P < 0.01) in pigs fed 0.039% BET compared with pigs fed 0.05% MET. In Exp. 4, a 2 x 2 x 2 factorial arrangement of treatments was used (MET, 0 or 0.072%; cystine, 0 or 0.059%; or BET, 0 or 0.057%). Overall ADG and gain:feed were increased (P < 0.10) in pigs fed MET. The intake of TSAA was increased (P < 0.05), and PUN was decreased (P < 0.10) in pigs fed MET or cystine. Overall ADFI was increased in pigs fed BET or MET independently but not affected when BET and MET were fed together (BET x MET, P < 0.10). The addition of BET to TSAA-deficient diets resulted in increased ADG, which was due to an increase in ADFI (TSAA intake). Thus, BET did not spare MET in this experiment. PMID- 11424695 TI - Use of growth hormone response to growth hormone-releasing hormone to determine growth potential in beef heifers. AB - The response of GH to GHRH at weaning is known to predict postweaning growth and body composition in beef bulls. The objective of this study was to determine whether GH response to a challenge of GHRH and plasma IGF-I can predict growth rate and body composition in the beef heifer. Growth hormone response to a challenge with two doses of GHRH was measured in 67 Angus heifers averaging 225 d of age (SD = 21) and 217 kg BW (SD = 32). Blood samples were collected at 0 and 10 min relative to an initial "clearance dose" (4.5 micrograms GHRH/100 kg BW) and again, 3 h later, relative to a challenge dose (1.5 or 4.5 micrograms GHRH/100 kg BW). Each animal received each of the two challenge doses, which were randomly assigned across 2 d of blood collection. Serum GH concentration was measured by RIA. Plasma was collected every 28 d during a 140-d growth test and assayed for IGF-I by RIA. Body weight was measured every 28 d and hip height was measured at weaning and at the end of a 140-d growth test. Average daily gain was calculated on d 140 of the growth test and body composition measurements were estimated by ultrasound 2 wk after completion of the growth test. Responses to the two GHRH challenges were dose-dependent (P < 0.05). Average daily gain tended to be related to GH response to the 1.5 micrograms GHRH/100 kg BW dose (R2 = 0.05; P = 0.06), but no relationship was observed at the 4.5 micrograms GHRH/100 kg BW dose (R2 = 0.00; P = 0.93). An inverse relationship (R2 = 0.06; P = 0.02) was observed between response to the 1.5 micrograms GHRH/100 kg BW dose and intramuscular fat percentage. Mean plasma IGF-I concentration was positively associated with ADG (R2 = 0.06; P < 0.01). Growth hormone response to GHRH is modestly related to body composition but not to ADG in weanling beef heifers and likely has limited use in evaluation of growth performance in replacement beef heifers. PMID- 11424696 TI - Effect of intracerebroventricular orexin-B on food intake in sheep. AB - Orexin is a hypothalamic neuropeptide that regulates feeding behavior in rats. Orexin-B has recently been cloned in pigs and was shown to stimulate food intake after intramuscular injection. This study was designed to determine whether intracerebroventricular (ICV) and intravenous injections of orexin could regulate appetite in sheep. Suffolk wethers were moved to indoor facilities, adapted to diets for 6 wk, and trained to stand in stanchions for 3 to 6 h each day for 2 wk before indwelling ICV cannulas were installed. These sheep were provided water and they consumed feed ad libitum. On the day before an experiment, each sheep was cannulated in a jugular vein. On the day of an experiment, sheep were placed in stanchions and allowed to stand for 1 h before use. Sheep were then monitored over a 2-h control period before i.v. injection with saline or porcine orexin-B (3 micrograms/kg BW) or ICV injection with artificial cerebrospinal fluid (CSF), orexin (0.03, 0.3, or 3 micrograms/kg BW) or in a second experiment with either orexin B (0.03, 0.3, 3 micrograms/kg BW), neuropeptide-Y (NPY; 0.3 microgram/kg BW), or orexin plus NPY. Food intake was monitored for consecutive 2-h periods. The i.v. injections of orexin did not affect food intake or metabolite or hormone concentrations. In ICV sheep, orexin increased food intake at 2 (P < 0.04) and at 4 h (P < 0.02). Food intake was greatest with the 0.3 microgram/kg BW dosage of orexin (P < 0.05). In the first 2 h after injection, orexin had an effect similar to that of NPY (0.23 kg for orexin and 0.2 kg for NPY). The combination of NPY and orexin had a greater effect on food intake (to 0.34 kg) than did either orexin (P < 0.05) or NPY (P < 0.008) alone. Differences were not apparent in the subsequent 2-h interval. No differences were noted in free fatty acid, glucose, growth hormone, luteinizing hormone, or insulin concentrations following orexin injection. There was an effect of ICV orexin treatment on plasma cortisol concentrations (P < 0.002). Cortisol was increased by orexin at the 0- to 2-h (P < 0.008) and in the 2- to 4-h (P < 0.009) intervals after orexin injection. These data indicate that central administration of orexin stimulates feed intake in sheep. PMID- 11424697 TI - Development of porcine embryos in vitro: effects of culture medium and donor age. AB - We compared development of porcine embryos in three media and evaluated the effect of age of the donor on embryo development in vitro. In Exp. 1, embryos were collected from 35 postpubertal females on d 2 or 3 after onset of estrus. Embryos were cultured 144 h in Whitten's Medium (WM), North Carolina State University Medium-23 (NCSU-23), or Beltsville Embryo Culture Medium-3 (BECM-3) in 95% air: 5% CO2 at 39 degrees C. More (P < 0.01) embryos that were initially one cell or two cells developed to blastocysts when cultured in NCSU-23 (56%) and BECM-3 (43%) rather than in WM (7.5%). More (P < 0.01) embryos that were four cells at recovery developed to blastocysts in NCSU-23 (97%) than in BECM-3 (69%) or WM (69%). Blastocysts that developed from four-cell embryos cultured in BECM-3 had more (P < 0.01) nuclei than blastocysts that developed from four-cell embryos in the other two media. In Exp. 2, ovarian responses, fertilization rates, and in vitro embryo development in NCSU-23 and BECM-3 were compared for postpubertal (approximately 170-d-old) gilts vs gilts given exogenous gonadotropins at 102 d of age. Ovulation rate (P < 0.01), number of eggs recovered, and number of eggs fertilized per gilt (P < 0.001) were greater in the older gilts. The percentage of eggs fertilized, the number of unfertilized eggs, and the number of unclassifiable eggs were similar (P > 0.10) for both age groups. More (P < 0.10) blastocysts developed from embryos recovered from 170-d-old than from 102-d-old gilts, and more (P < 0.05) blastocysts developed in NCSU-23 than in BECM-3. Zona thicknesses and number of nuclei per embryo were similar (P > 0.10) for both ages. We conclude that embryos from prepubertal gilts do not have the same in vitro developmental potential as those from cyclic gilts. However, superior development of embryos in NCSU-23 from both 102-d-old and 170-d-old gilts indicates that media composition did not differentially affect embryos produced by younger vs older gilts. PMID- 11424698 TI - A mechanistic model of whole-tract digestion and methanogenesis in the lactating dairy cow: model development, evaluation, and application. AB - Dietary intervention to reduce methane emissions from lactating dairy cattle is both environmentally and nutritionally desirable due to the importance of methane as a causative agent in global warming and as a significant loss of feed energy. Reliable prediction systems for methane production over a range of dietary inputs could be used to develop novel dietary regimes for the limitation of feed energy loss to methane. This investigation builds on previous attempts at modeling methanogenesis and involves the development of a dynamic mechanistic model of wholerumen function. The model incorporates modifications to certain ruminal fermentation parameters and the addition of a postruminal digestive element. Regression analysis showed good agreement between observed and predicted results for experimental data taken from the literature (r2 = 0.76, root mean square prediction error = 15.4%). Evaluation of model predictions for experimental observations from five calorimetry studies (67 observations) with lactating dairy cows at the Centre for Dairy Research, in Reading, U.K., shows an underprediction (2.1 MJ/d) of methane production (r2 = 0.46, root mean square prediction error = 12.4%). Application of the model to develop diets for minimizing methanogenesis indicated a need to limit the ratio of lipogenic to glucogenic VFA in the rumen and hindgut. This may be achieved by replacing soluble sugars in the concentrate with starch or substituting corn silage for grass silage. On a herd basis, the model predicted that increasing dietary energy intake per cow can minimize the annual loss of feed energy through methane production. The mechanistic model is a valuable tool for predicting methane emissions from dairy cows. PMID- 11424699 TI - Intake restriction strategies and sources of energy and protein during the growing period affect nutrient disappearance, feedlot performance, and carcass characteristics of crossbred heifers. AB - Two trials were conducted to evaluate intake restriction, energy, and protein source on the performance and carcass merit of heifers limit-fed corn gluten feed. Trial 1 crossbred heifers (n = 140) were allotted to ad libitum wet corn gluten feed (WCGF)-hay, ad libitum WCGF-corn, WCGF-corn continuously limit-fed to achieve a gain of 1.1 kg/d or WCGF-corn fed in the following sequence: 70% of ad libitum for 20 d, ad libitum for 20 d, and 2 d common intake during the 84-d growing period. One-half of these restricted-refed heifers received a rumen undegraded protein supplement, and the other half received a rumen degraded protein supplement. Heifers were fed a common finishing diet ad libitum subsequent to the growing period. Heifers offered ad libitum WCGF-hay and WCGF corn were slaughtered at a common compositional fat end point. Heifers limit-fed WCGF-corn were slaughtered at the same time as heifers offered ad libitum WCGF corn, regardless of subcutaneous fat cover. Limit-feeding WCGF-corn diets to growing heifers reduced ADG (P < or = 0.01) but did not compromise feed efficiency. Method of intake restriction, continuous or ad libitum-interrupted, and supplemental protein source did not affect combined growing-finishing performance. However, when fed for a common length of time, the average of the limit-fed heifers had lighter carcass weights (P < or = 0.01) and lower (P = 0.04) marbling scores. Heifers offered ad libitum WCGF-hay gained slower (P < or = 0.01) and less efficiently (P < or = 0.01) than heifers offered WCGF-corn. Trial 2 crossbred heifers (n = 222) were allotted to dry corn gluten feed (DCGF) corn ad libitum or restricted to 80% of ad libitum for 42, 84, or 126 d; or ad libitum corn silage or DCGF-corn silage at 80% of ad libitum for 84 d. Feed efficiency (P = 0.07) and ADG (P = 0.08) tended to behave quadratically, being poorer for heifers limit-fed for 126 d during the growing period. Heifers limit fed DCGF-corn gained more efficiently (P = 0.05) than heifers grown on ad libitum corn silage. Limit-feeding CGF-corn to growing beef heifers can be used to achieve moderate rates of gain without compromising feed efficiency. However, limit-feeding during the growing period may result in lighter weight carcasses with lower quality grades if not fed to the same fat end point as heifers grown with free access to concentrate. PMID- 11424700 TI - Effect of heterogeneous digesta chemical composition on the accuracy of measurements of fiber flow in dairy cows. AB - The objective of the current study was to evaluate the effect of unrepresentative sampling of digesta particulate matter entering the omasal canal on the accuracy of fiber flow measurements. The experimental design comprised one period, one diet, and three cows as experimental units. Within each cow, the physical and chemical composition of digesta particulate matter was assessed at seven sites within the digestive tract. Three Finnish Ayshire dairy cows, equipped with ruminal and simple-T duodenal cannulas, in extended lactation were offered grass silage twice daily on an ad libitum basis. Digesta samples were collected from the rumen (dorsal and ventral sac), reticulum, omasal canal, omasum, duodenum, and rectum to determine particle size distribution in digesta, chemical composition of various particle size fractions, and distribution of two flow markers (Cr-labeled straw and indigestible NDF [INDF]) among particle size fractions. Digesta samples were wet-sieved using sieves of 2.50, 1.25, 0.630, 0.315, 0.160, and 0.080 mm. Particulate matter was analyzed for OM, NDF, and Cr concentrations, and INDF concentration was determined based on 12-d ruminal incubation. The particle size of digesta entering the omasal canal was larger compared with the omasum or the duodenum, suggesting that omasal canal samples were not representative of particle size distribution truly escaping the rumen. The concentration of potentially digestible NDF (PDNDF) decreased with decreasing particle size. The PDNDF concentration of particulate matter retained on all sieves was greatest in the rumen and gradually decreased along the digestive tract. From the reticulorumen to the omasum, the decrease was associated with decreased particle size, reflecting selective passage of particulate matter. In contrast, from the omasum to the duodenum and rectum, the PDNDF concentration decreased within each particle size fraction without effect on particle size, indicating a nonselective passage of particulate matter between these sites. Variation between particle size fractions was slightly greater for Cr concentration than for INDF concentration, indicating that unrepresentative sampling of particulate matter had a greater effect on Cr concentration compared with that of INDF. Owing to unrepresentative sampling, NDF entering the omasal canal was overestimated by 5% using INDF and underestimated by 7% using Cr as a particle phase marker. Of total NDF digestibility, proportionally 0.90, 0.07, and 0.03 occurred in the reticulorumen, omasum, and intestines, respectively. The current results indicate that, despite unrepresentative sampling of digesta particulate matter entering the omasal canal, the errors in determined NDF flow were small. The omasum may have a greater role in postruminal NDF digestion than the intestines. PMID- 11424701 TI - Resistance of feed enzymes to proteolytic inactivation by rumen microorganisms and gastrointestinal proteases. AB - Potential feed enzyme additives for ruminants were tested in vitro for their stability to ruminal microbial and gastrointestinal proteolysis. Four commercial preparations from Trichoderma longibrachiatum (A, B, C, and D) and one from an undisclosed source (E) were incubated up to 6 h with ruminal fluid taken from four lactating dairy cows before or 2 h after feeding. The stability of preparation B was also tested in the presence of pepsin at pH 3 and pancreatin at pH 7. Cellulase (EC 3.2.1.4), cellulose 1,4-beta-cellobiosidase (EC 3.2.1.91), beta-glucanase (EC 3.2.1.6), xylanase (EC 3.2.1.8), beta-glucosidase (EC 3.2.1.21), and beta-xylosidase (EC 3.2.1.37) activities were monitored throughout the incubations. Polysaccharidase activities of all enzyme preparations were remarkably stable in ruminal fluid taken after feeding. Ruminal fluid obtained before feeding inactivated the polysaccharidases in preparations B and D to a greater extent than ruminal fluid obtained after feeding. Cellulase and cellulose 1,4-beta-cellobiosidase activities were the least stable, declining (P < 0.05) by 35 and 60% for preparations B and D, respectively. Xylanase activity of preparation D decreased (P < 0.05) by up to 30% after 6 h of incubation, whereas beta-glucanase activity was not affected. The ability to degrade exogenous enzymes also differed among cows (P < 0.05). Pepsin and acid (pH 3.0) did not affect polysaccharidases in preparation B but decreased glycosidase activities by 10 to 15% (P < 0.05) after 1 h of incubation. Pancreatin, at the maximum concentration used, inactivated cellulase, cellulose 1,4-beta-cellobiosidase, and xylanase activities at a rate of 0.55, 1, and 0.45%/min, respectively. beta Glucosidase and beta-xylosidase activities decreased by 1 and 0.75%/min, respectively. Partial proteolysis of cellulase, cellulose 1,4-beta cellobiosidase, and xylanase by pancreatin produced a transient increase in activity. This twofold increase for cellulase and fourfold increase for cellulose 1,4-beta-cellobiosidase was directly proportional to pancreatin concentration. These results suggest that the enzyme feed additives tested were stable in the rumen of animals after feeding. Exogenous enzymes are likely to be more susceptible to the host gastrointestinal proteases in the abomasum and intestines than to ruminal proteases. However, exogenous polysaccharidases may survive for a considerable period of time in the small intestine and they probably maintain activity against target substrates in this environment. PMID- 11424702 TI - Rapid communication: linkage mapping of seven bovine cDNA. PMID- 11424703 TI - Rapid communication: genetic linkage and physical mapping of the porcine phospholipid transfer protein (PLTP) gene. PMID- 11424704 TI - [20th consensus conference on intensive care and emergency medicine (Corticosteroids in the treatment of septic shock and respiratory distress syndrome]. PMID- 11424705 TI - [Antibiotics and lower respiratory tract infections: the message is not getting through!]. PMID- 11424706 TI - [Age is not a formal or relative risk factor for lung cancer surgery]. PMID- 11424707 TI - [Making the decision to hospitalize and mechanically ventilate cancer patients. Towards a new sequential concerted approach?]. PMID- 11424708 TI - [Music and respiratory pathology]. AB - Musical performance, especially in singers and wind instrument players, depends on an effective pulmonary function. Performing artists may be seriously impaired by respiratory diseases that, comparatively, may produce only modest inconvenience for non-musicians. The report of two cases of respiratory diseases occurring in musicians herein provides an introduction to a review of the interactions between music and the human respiratory system. The following points are considered: epidemiological data; pulmonary function in musicians; favorable effects of music on the respiratory system; description of the main respiratory problems that may affect musicians. PMID- 11424709 TI - [Respiratory infections during chemotherapy-induced aplasia]. AB - Damage to local and systemic host defenses of the lung makes the immunocompromised patient vulnerable to inhaled microorganisms. When a pulmonary infiltrate occurs, the array of possibilities is very large including conventional and opportunistic agents. The type of underlying disease and its associated immunodeficiency allow a high degree of accurate pathogen prediction. Neutropenia is associated with Gram-negative bacilli pneumonia. Prolonged neutropenia increases the risk of invasive aspergillosis and other unusual mycotic agents. Cellular immunodeficiency is associated with intracellular microorganisms including Mycobacteria spp., Nocardia spp., Legionella spp., Rhodococcus equi, cytomegalovirus, Strongyloides stercoralis, Toxoplasma gondii, Histoplasma capsulatum, Coccidioides spp., Cryptococcus neoformans and Pneumocystis carinii, parasites such as Toxoplasma gondii and Strongyloides stercoralis, and virus such as cytomegalovirus, Herpes simplex or zoster, adenovirus, respiratory syncitial virus and measles. Humoral immunodeficiency predisposes to infection with encapsulated pathogens such as S. pneumoniae and Haemophilus influenzae. Chest computerized tomography scan and bronchoalveolar lavage are essential procedures for diagnosis. However, despite continuous progress in diagnostic methods, the specific etiology remains often unknown. Successful treatment depends on the type of pathogen, status of host defences and early adequate choice of antibiotic. Enhancement of host defences with growth factors and cytokines may decrease the incidence and improve the final outcome of respiratory infections in the immunocompromised host. PMID- 11424710 TI - [Mechanical ventilation in the cancer patient]. AB - The overall objective of this review is to analyze the literature published on artificial ventilation in cancer patients. First, we reviewed series of cancer patients treated by mechanical ventilation or intensive care series with data on artificial ventilation in a context of malignant solid tumors, hematology malignancies or complications of bone marrow transplantation. A second part of the review is devoted to characteristic diseases for which specific literature is available, with description of the clinical presentation, the causes and the proposed therapies. The third part is a discussion of considerations on care and therapeutic techniques. Finally, we propose guidelines for a practical approach to this type of patient taking into account the prognosis and new techniques such as noninvasive ventilation. PMID- 11424711 TI - [Specific bronchial provocation test with solid aerosols. Quantification of results]. AB - OBJECTIVES: To determine 1) the level of specific bronchial reactivity by challenge with flour and 2) the criteria of positivity using a new method for the expression of the results. METHOD: Thirty-eight asthmatic subjects, occupationally exposed to wheat flour, performed a challenge with lactose then with flour. The instantaneous measurement of the concentration and of the inspiratory airflow were used to calculate the inhaled dose and to establish the dose-response relationships. The results were given 1) by the dose of flour provoking a 20% fall in FEV1 (PDf20), 2) by the comparison of the variation of the FEV1 during the challenge with flour to the distribution of the values observed during the challenge with lactose (inferior limit of the confidence interval at 99.7%). RESULTS: The variations of the FEV1 were not significantly related to the inhaled dose of lactose. The specific bronchial reactivity to flour was a continuous data and three groups were distinguished: 1) subjects (n = 15) with high bronchial reactivity had a fall of FEV1 of more than 20% 2) subjects (n = 13) without significant variation of the FEV1 for doses higher than 1,400 micrograms by comparison to the distribution of the values of the lactose test 3) subjects (n = 10) with a significant fall of FEV1 by comparison to the distribution of the values of the lactose test but lower than 20%. For this group with moderate reactivity, the flow of the inhaled dose may be determinant for the bronchial response. CONCLUSIONS: PDf 20 measures the specific bronchial reactivity. However, if the fall in FEV1 is lower than 20%, the specific challenge with flour may be compared to the challenge with lactose to detect the subjects with moderate reactivity. Our results confirmed the role of the inhaled dose and suggested the role of the dose rate in the outset of bronchial obstruction among asthmatic subjects. PMID- 11424712 TI - [Management of lower respiratory tract infections by general practitioners in France]. AB - OBJECTIVE: The purpose of this study was to describe the diagnostic and therapeutic strategies used by general practitioners (GPs) in patients with lower respiratory tract infections (LRTI). METHODS: Four hundred fifty GPs practicing in France participated in the study; they included 804 patients. The GP recorded social and demographic data and their prescription on a data sheet and responded to a phone questionnaire about their strategy. RESULTS: Most of the LRTI were acute bronchitis (72%); pneumonia and acute exacerbations of chronic bronchitis were observed respectively in 11% of the patients recruited. Diagnostic criteria used by the GPs were generally auscultation signs in patients with fever, cough and expectoration. Specialized advice (always a chest physician) and hospitalization were exceptional. Prescription of complementary exams was strongly related to the diagnosis of pneumonia (OR = 33.3; CI0.95: 15.48-70.4). Sick leaves were related to general symptoms (fever, asthenia). Antibiotics were prescribed in 95.7% of the patients, mainly aminopenicillin (40.4%) and macrolides (33.2%). Nonsteroidal or steroidal antiinflammatory drugs were prescribed in 72.5% of the patients irrespective of the LRTI diagnosis. DISCUSSION: The fact that GPs do not often refer patients to specialists or order hospitalization confirms their important role in setting up recommendations. It would also be necessary to develop an education program on better use of antibiotics targeted to GPs and patients. PMID- 11424713 TI - [Non-small cell lung cancer: surgical trends as a function of age]. AB - Most of the studies on lung cancer and age are usually done on very young or very old populations. We conducted a study of the evolution of surgical features over time in a population aged 24 to 89 years. The series included 1,809 men and 287 women (n = 2,096) who had undergone surgery between April 1984 and December 1990 (n = 1,026) and between January 1991 and December 1996 (n = 1,996). Patients were divided into 7 age groups with 2 subgroups for patients with or without prior medical history of cancer. We analyzed the type of surgery and pathology findings by age. Morbidity was recorded according to state of previous cardiovascular disease and long-term survival was analyzed. A significant increase in the number of female patients was noted during the last ten years. Adenocarcinomas were more frequent in young patients. Exploratory interventions, partial tumor resections and lung resections for metastasis were more frequently performed in young patients than in older patients. Excision of mediastinal nodes was less performed often in old patients. In case of curative resection, postoperative pTNM was not modified with age. Morbidity increased with age; mortality was more frequent in the elderly even when comorbidity was taken into account. Mortality was not related significantly with cardiovascular morbidity factors despite an increased frequency of previous cardiovascular disease with age. Survival according to age showed 3 main types of population: patients aged under 64 years, those between 65 and 74, and those over 75 years of age. Mortality increased with time but was less often related to recurrence of lung cancer. Nevertheless, survival for stage III and stage IV patients older than 75 years demonstrated that surgery was not indicated for these patients. This study shows that indications for surgery should not vary with age except for patients over 75 years who have locally advanced cancer and a risk of surgical death greater than the chances of survival. All non-small-cell lung cancers should be resected with no delay, even in the elderly population. PMID- 11424714 TI - [Pseudotumoral forms of sarcoidosis]. AB - We report 5 cases of sarcoidosis in 4 men and 1 woman who presented multinodular pulmonary lesions. Seldom described (1 to 4% of all cases of sarcoidosis), a multinodular pulmonary presentation is suggestive of metastatic disease. In our patients, the parenchymal opacities were multiple, peripheral and exhibited fuzzy limits, measuring 1 to 7 cm in size and frequently associated with mediastinal adenopathies. The radiographic pattern contrasted with the clinical manifestations (3 of the 5 cases were fortuitous discoveries). Pathological proof required to rule out other disease, especially tumor formation, was acquired; mediastinoscopy allowing the diagnosis in 3 out of 5 cases. The clinical course was favorable in all cases without treatment within 8 +/- 5 months and a mean follow-up of 5 years (range 9 months to 14 years). PMID- 11424715 TI - [Congenital esophagobronchial fistula in an adult]. AB - Congenital esophagobronchial fistulae are rare and are most often observed in neonatalogy. We present the case of a 21-year-old man with a history of chronic bronchorrhea due to middle and lower lung lobe destruction. This case demonstrates the diagnostic features in individuals with unexplained suppurating respiratory symptoms and the difficulty in identifying the congenital pathogenesis of this type of fistula. PMID- 11424716 TI - [Sinobronchial syndrome]. AB - The sinobronchial syndrome consists of chronic sinusitis and chronic nonspecific inflammation of the lower airways, typically chronic bronchitis, bronchiectasis and diffuse panbronchiolitis. This airway disease of unknown etiology is not related to smoking. It is common in Japan and is poorly described in western countries. We report a case in a non-Asian patient. The known efficacy of long term low-dose erythromycin therapy justifies careful diagnosis of sinobronchial syndrome in patients of non-Asian ethnic origin. PMID- 11424717 TI - [Small cell lung cancer revealed by extramembranous glomerulonephritis]. AB - We describe a case of small-cell lung cancer limited to the thorax but with malignant pleural effusion in a 47-year-old man that was revealed by a nephrotic syndrome due to membranous glomerulonephritis (MGN). Chemotherapy led to a partial tumor response with total resolution of the nephrotic syndrome. Tumor relapse did not provoke proteinuria. Primary lung cancer is the cause of about 3% of all cases of MGN and 40% of tumor-related MGN. There are 49 cases of tumor related MGN in the literature, including 9 cases of small-cell lung cancer. PMID- 11424718 TI - [Respiratory bronchiolitis with diffuse interstitial lung disease]. AB - Respiratory bronchiolitis-associated interstitial lung disease (RB-ILD) is a disease that exclusively affects cigarette smokers. Long-term prognosis is good with cessation of smoking, in combination or not with corticosteroid therapy. We report here the case of a 50-year-old patient with RB-IL diagnosed on lung biopsy. Despite corticosteroid and cyclophosphamide therapy, no functional or radiological improvement was obtained. In contrast, cessation of smoking was associated with the disappearance of the infiltrative opacities. Clinical and radiological parameters remained stable during follow-up (13 years) while a moderate obstructive pattern appeared. PMID- 11424719 TI - [Pseudoneoplastic forms of bronchiolitis obliterans with organizing pneumopathy (BOOP)]. AB - Chronic obstructive bronchiolitis with organizing lung disease is an anatomoradioclinical entity characterized by nonspecific inflammation associated with recurrent migratory minimally dense alveolar opacities on the chest x-ray poorly responsive to corticosteroid therapy. Excepting this typical presentation, other clinical forms may occur. We report the cases of two patients with an exceptional localized presentation raising the differential diagnosis of lung cancer. PMID- 11424720 TI - [Air trapped in the right upper lobe]. PMID- 11424721 TI - The gift of life. PMID- 11424722 TI - Early and periodic screening, diagnosis, and treatment examination completion rates for Oklahoma Medicaid managed care: 1995-1998. AB - Although quality medical care is a goal of all health care providers, finding a means by which to take that idea from the abstract to the measurable is often an arduous task. The Oklahoma Health Care Authority and the Oklahoma Foundation for Medical Quality confronted that challenge when examining the state of Oklahoma's Early and Periodic Screening, Diagnosis, and Treatment (EPSDT) examination completion rates. Focused on measuring processes of care, the Oklahoma Health Care Authority (OHCA) and the Oklahoma Foundation for Medical Quality (OFMQ) used chart abstraction to track EPSDT examination rates for SoonerCare Plus Medicaid managed care recipients from 1995 through 1998. During the abstractions, an examination was interpreted as an EPSDT screen if a comprehensive health and development history plus a comprehensive unclothed physical examination were recorded. Examination rates have improved from 1995 when there was a documented 17.6% completion rate. The 1998 documented rate of completion was 60.0%. Trend analysis shows significant improvement over the four-year period. The results also suggest the necessity for continued improvement in the provision of documented EPSDT examinations to individuals in Medicaid managed care plans in Oklahoma. PMID- 11424723 TI - The Human Genome Research Project: implications for the healthcare industry. AB - Diseases have ravaged the human race throughout history. Although medical science has progressed rapidly over the past few decades and increased the average life span from 56 to 84, the quest for a disease-free life process has been elusive. But the history of human biology was altered forever nearly 40 years ago by a bold decision to launch a research program effort to characterize in ultimate detail the complete set of genetic instructions of the human being. In 1988, Congress appropriated funds to the Department of Energy and the National Institutes of Health to begin planning the Human Genome Project. Planners set a 15-year time frame, estimated that the price tag would be $3 billion, and laid formal goals to get the job done. On October 1, 1990, the Human Genome Project officially began. The Project aimed that by 2005, the "Holy Grail" of life would be deciphered. This revelation would provide a new understanding of the genetic contributions to human diseases and help in the development of rational strategies to minimize or prevent diseases in the future. PMID- 11424724 TI - Child safety seats for the prevention of traffic injuries. AB - It's an old joke among new parents in Labor and Delivery: How many people did it take to install your car seat? Although the remark elicits chuckles from new moms and dads, using a child safety seat--and using it correctly--is no laughing matter. More children between the ages of one and ten die in automobile crashes than from any other type of injury or disease. In the under-one-year age group, this health risk is second only to Sudden Infant Death Syndrome (SIDS). Some physicians, knowing how complicated child safety seats can be to use, may be reluctant to give advice to parents on their use. In addition, the potential liability of dispensing incorrect information may be enough to make some doctors reluctant to do so. However, there are some basic tips that physicians can and should be passing on to parents. Most parents place great trust in their child's physician; asking about the child's transportation and advocating proper child safety seat use is an effective prevention technique. PMID- 11424725 TI - Analysis of material and energy flow in sewage treatment facilities in Japan. AB - Energy consumption in sewage treatment facilities in Japan has increased due to increasing tap water consumption. To reduce the resource/energy consumption in sewage treatment facilities, measures such as the selection of optimum treatment processes and operating conditions should be considered. The objective of this study is to gather information necessary for the determination of optimum sewage treatment processes and optimum operating conditions. The energy consumption and material flow in sewage treatment facilities in Japan are analyzed using statistical data. In 1994, reuse rate of treated sewage outside the treatment facilities in Japan was 18% of the amount of domestic treated water. In this regard, reuse of water outside facilities should be encouraged. Average electric power consumption per unit volume of wastewater in sewage treatment facilities varies widely from facility to facility and closely correlates with the facility scale. For example, the smaller the facility scale, the larger the electric power consumption. Treatment volume of sewage in smaller facilities is much less than their capacity. 3.7 million t year-1 of dehydration cake is incinerated and 0.1 million t year-1 of it is converted by composting. The recycle rate of the cake was low. Developing a new sludge treatment process other than incineration is necessary. PMID- 11424726 TI - Evaluation of Activated Sludge Model No. 2 at high phosphorus concentrations. AB - This paper presents laboratory scale experimentation carried out to study enhanced biological phosphorus removal at high phosphorus concentrations in a sequencing batch reactor. Four series of data obtained in a sequencing batch reactor are examined in light of the Activated Sludge Model No. 2. This model was calibrated using data from the first and second series working at low phosphorus concentrations. The Activated Sludge Model No. 2 successfully characterised the enhanced biological phosphorus removal performance of the sequencing batch reactor at low phosphorus concentrations. The calibrated model was then used to adjust experimental results of the other series working at high phosphorus concentration. Differences between model predictions and experimental data could be explained by redissolution in the anaerobic phase and precipitation in the aerobic phase of calcium phosphates not taken into account by the model. This hypothesis can be justified by the conditions of pH and phosphorus concentration prevailing in the experiments and it was confirmed by the results of another experiment in which precipitated phosphorus profile was measured during the cycle. PMID- 11424727 TI - Biosorption and desorption of cadmium(II) by biomass of Laminaria japonica. AB - Biosorption and desorption properties of cadmium(II) from aqueous solutions by the biomass of marine alga Laminaria japonica were investigated. Results indicated that the uptake capacities were solution pH dependent and a maximum uptake capacity of about 1.3 mmol g-1 (dry weight) was observed at pH 6. The adsorbed cadmium cannot be desorbed by distilled water, but it can be effectively recovered by using acidic or EDTA solutions. The equilibrium isotherms can be described well with the Langmuir adsorption equation. Biomass, pre-treated with calcium solution exhibited a higher (about 30%) uptake capacity and can be easily settled from aqueous solutions. Batch kinetics experiments indicated that more than 90% of the adsorption occurred within 20 minutes of agitation and equilibrium was reached within one hour. Fixed-bed experiments showed similar uptake capacities to those of batch results and sharp breakthrough curves were obtained. This study indicated that the biomass of L. japonica can be used as an efficient biosorbent for the removal and recovery of cadmium(II) from waste water streams. PMID- 11424728 TI - The kinetics of catalytic incineration of C2H5SH and (CH3)2S2 over a Pt/Al2O3 catalyst. AB - Catalytic incineration is one of the cost-effective technologies to solve the troublesome VOCs. However, some sulfur containing VOCs, such as ethyl mercaptan and dimethyl disulfide, may deactivate the Pt catalyst that is commonly used in the catalytic incineration process. The catalytic incineration of these compounds over a Pt/Al2O3 catalyst was carried out in a bench scale catalytic incinerator. Three kinetic models, such as power-rate law, Mars and Van Krevelen model, and Langmuir-Hinshelwood model were used to analyze the results. A differential reactor design was used for best fit of kinetic models in this study. The results show that the Langmuir-Hinshelwood model is feasible to describe the catalytic incineration of both C2H5SH and (CH3)2S2. This suggests that the chemical adsorption of O2 molecule is important in the process of catalytic incineration of C2H5SH and (CH3)2S2. PMID- 11424729 TI - Pre-evaporative fenton remediation of treated municipal wastewater for reuse purposes. AB - The present study explored the application of evaporative technology as an alternative desalination technique for wastewater treatment plant effluent. In the experiments carried out, it was shown that evaporation allowed for extensive decontamination of wastewater treatment plant effluent as most inorganic and organic contaminants were completely removed. Ammonia could be retained in the concentrate when pH was adjusted to values lower than 4; however this led to increased nitrite volatility due to the formation of nitric oxide. Using an electronic nose, it was observed that evaporation gave rise to a change in odour quality and quantity due to volatilisation of the volatile organic compounds present in the effluent. Remediation of these components was carried out by Fenton oxidation prior to evaporation. A high removal was observed under the applied conditions of Fe2+ = 25 mg l-1, H2O2 = 50 mg l-1, pH = 3 and a reaction time of 2 hours. PMID- 11424730 TI - Study on sludge expansion during treatment of salad oil manufacturing wastewater by yeast. AB - Five yeast strains, namely Rhodotorula rubra, Candida tropicalis, Candida utilis, Candida boidinii, Trichosporon cutaneum, were isolated from soil spots of a salad oil factory, and applied for continuous treatment of salad oil manufacturing wastewater. The oil and COD removal performance of the mixed cultures were comparable to the results other researchers obtained. Sludge expansion, accompanied with sludge morphology change from pseudomycelia to true mycelia, occurred during continuous treatment of wastewater. The true mycelia dominated sludge had a much higher water content and SVI value than that of the yeast pure cultures, although the two kinds of sludge had similar oil removal performance. A mold, Geotrichum candidum, was isolated from the expanded sludge, and was suspected to be a reason for sludge expansion. Addition of 0.3% sodium propionate into batch cultures degraded SVI value from around 100 to 60. In a continuous running, addition of 10 mg l-1 sodium hypochlorite decreased SVI value from over 200 to below 100. The yeast activity, however, was weakened to a large extent at the same time. PMID- 11424731 TI - Occurrence of antifouling biocides in the Spanish Mediterranean marine environment. AB - A compilation of the results of a monitoring program of the recently used antifouling pesticides diuron (3-(3,4-dichlorophenyl)-1,1-dimethylurea), Irgarol 1051 (2-methylthio-4-tertiary-butylamino-6-cyclopropylamino-s-teiazine), seanine 211 (4,5-dichloro-2-n-octyl-4-isothazolin-3-one), chlorothalonil (2,4,5,6 tetrachloro-isophthalonitrile), dichlofluanid (N'-dimethyl-N-phenylsulphanamide), TCMTB ((2-thiocyanomethylthio) benzothiazole), and three degradation products demethyldiuron (3-(3,4-dichlorophenyl)-1-methylurea),3,4-dichlorophenylurea and 2 methylthio-4-tert-butylamino-s-triazine (Irgarol degradation product) that was carried out between April 1996 and February 2000 in enclosed seawaters from Catalonia and Almeria (Spanish Mediterranean coast) is reported. Nine points were sampled along the Catalan coast: Barcelona Olympic port, Masnou, Blanes, Sant Carles de la Rapita, Tarragona, Cambrils and Salou marinas as well as the Cambrils and Tarragona fishing harbors and in marinas and ports from Almeria: Aguadulce port, Almeria port, Almerimar fishing harbour and Almerimar marina. The analytical methodologies were based on Solid Phase Extraction followed by liquid chromatography (LC) or gas chromatography (GC) coupled to a mass spectrometry (MS) or -Diode Array Detector. The main pollutants found in the sampled points were diuron and Irgarol 1051 that were detected at concentrations up to 2.19 micrograms l-1 and 0.33 microgram l-1, respectively. On the other hand, seanine 211 was found at the highest concentration (up to 3.7 micrograms l-1) during the summer of 1999. Low concentrations of dichlofluanid and the above mentioned degradation products were detected for the first time in the Spanish coasts. Chlorothalonil, TCMTB were not found at concentrations higher than 1 and 20 ng l 1 respectively which were the limit of determination (LOD) of the method for these compounds. In general the contamination at the different marinas is higher at the end of spring and in summer where the boating activity is also higher. This paper shows for the first time that the contamination by the new antifouling pesticides in Spanish coastline, basically marinas and fishing harbours, is permanent along the whole calendar year. So, preventive actions by the harbour authorities will be needed in the near future in order to monitor and control the levels and effects of the new antifouling biocides in the marine environment. PMID- 11424732 TI - Nitrogen removal from sewage by continuous flow SBR system with intermittent aeration. AB - A continuous-flow sequential batch reactor system with intermittent aeration known as the intermittent cycle extended aeration system was used to investigate the efficiencies of nitrogen removal for sewage. In the conventional intermittent cycle extend aeration system with continuous aeration process to treat the synthetic sewage wastewater under 3 cycles per day (cycle time = 8 hours), high COD removal efficiencies were achieved, but high levels of nitrate (22.3-23.6 mg l-1) and decreased pH values were also found in the effluent. In the intermittent aeration, the dissolved oxygen in the reactor was reduced from the range of 3.5 5.0 mg l-1 to 0.5-1.2 mg l-1, the nitrogen in the influent was partially oxidized to nitrite-nitrogen, although nitrate-nitrogen was also present in the effluent. In detailed cyclic study of the reactor with relatively low D.O. (0.3-1.0 mg l-1) and high MLSS (3200 mg l-1 at half full water level), the experimental results of intermittent aeration processes showed higher oxidation rates of ammonia to nitrite (1.44-3.55 mg N g-1 MLSS hr-1) than the conversion rates of nitrite to nitrate (0.00-1.22 mg N g-1 MLSS hr-1). It was thus, concluded that denitrification rates were not affected by the limited supplement of soluble organic carbon in the system. In addition, NOx(-)-N could be removed with a total nitrogen removal rate up to 94.9% when the MLSS was controlled in concentrations of 3200 mg l-1 at half full water level with low D.O. The process of intermittent aeration of ICEAS could provide an economic and effective means for nitrogen removal in treating sewage. PMID- 11424733 TI - Enhancement in the treatment efficiency and conversion to energy of dairy wastewaters by augmenting CST reactors with simple biofilm support systems. AB - Dairy wastewaters are rich in organic carbon and nutrients. When disposed without proper treatment they cause severe pollution of water and land. But if treated by anaerobic fermentation such wastewaters can yield useful clean energy in the form of methane. The systems commonly used for such fermentation throughout the world are Continuously Stirred Tank Reactors (CSTRs). We describe attempts to upgrade CSTR performance by incorporating biofilm support systems (BSSs) within the existing reactors. The focus of the work has been to find inexpensive and easy-to install BSS which could significantly enhance the rate of waste treatment as well as quantum of energy recovery (as methane). Our experiments with various BSS options have revealed that rolls of nylon mesh (-1 mm openings), of 5 cm height and 2 cm dia, when incorporated in CSTRs at the biofilm-surface: digester-volume ratio of 0.3 cm2 cm-3, enabled the CSTRs to perform better with up to 25% improvement in methane yield. The studies, thus, reveal that simple devices of the type used by us as BSS can significantly improve the performance of CST anaerobic digesters treating dairy wastes. The enhancement is evidently due to the development of active biofilms on the BSS which not only enhance the contact of the water with micro-organisms but also reduce microbial washout. Further, such devices are inexpensive and very easy to incorporate--the gains are thus achieved with very little cost and effort. BSSs were also found by us to enhance conversion of volatile fatty acids--derived earlier by us from aquatic weed water hyacinth--into methane-rich biogas. PMID- 11424734 TI - Determination of volatile organic compounds in emissions by coal-fired power stations from Spain. AB - This study concerns the emissions of volatile organic compounds (VOCs) by coal fired power stations. The main compounds are monoaromatic hydrocarbons and aliphatic hydrocarbons, chlorinated compounds have less importance. The influence of combustion parameters can not be clearly established. Emissions factors were calculated and they are smaller than those of other anthropogenic combustions. A comparative study of two sources of VOCs, power stations and motor vehicles, indicates that the environmental impact of the latter are most important. PMID- 11424735 TI - Characterisation and anaerobic batch degradation of materials accumulating in anaerobic digesters treating poultry slaughterhouse waste. AB - We characterised materials accumulating in two failed mesophilic semi-continuous anaerobic digesters treating poultry slaughterhouse waste and, for reference, materials in the two well-performing digesters, to find the anaerobic degradability of these materials and the factors affecting their degradation. We also studied materials accumulating and stratifying in various layers in one of the two well-performing digesters. The material from the most severely failed digesters produced methane sluggishly and did not improve appreciably even with 33 percent dilution suggesting that the recovery of failed process is slow. The methane production was apparently affected by the accumulated long-chain fatty acids, totalling 8.1 g l-1, which degraded slowly. However, the material produced methane in the end, which shows that the failure was reversible. In the well performing digester, considerable amounts of long-chain fatty acids already floated on top of the digester after 20 hours without mixing, a phenomenon which may have affected their bioavailability and toxicity. However, materials from the top, middle, and bottom layers of the digester were readily and largely methanised by the microbial populations present in them and additional inocula did not markedly enhance the methanation. The results indicate that long-chain fatty acids are apparently the main factor affecting both the failure and recovery of a poultry slaughterhouse waste digester. Thus excessive feeding of lipids into the digester should be avoided. PMID- 11424736 TI - Start-up performance of a subsurface-flow constructed wetland for domestic wastewater treatment. AB - Constructed wetlands have been shown to provide sufficient domestic wastewater treatment in temperate climates. However, the effectiveness of this technology, when subjected to severe temperature extremes during start-up conditions, is limited. Because of the growing interest in constructed wetland technology within smaller communities located in colder-climates, and the need for cost-effective treatment systems, this paper presents the performance of a constructed wetland system during the first two-years of operation under a variety of loading and operating conditions associated with northern climates. While the treatment performance of constructed wetland systems has been widely documented within the literature, documentation on performance during start-up stages has been limited. The results indicate that effective and sufficient constructed wetland seasonal removals of total suspended solids, volatile suspended solids, 5-day carbonaceous biochemical oxygen demand, chemical oxygen demand, and fecal coliform were achieved within the wetland cell during the first two-years of operation. Wastewater temperatures appeared to affect 5-day carbonaceous biochemical oxygen demand and chemical oxygen demand removal rates. Nitrogen and phosphorus reductions were not as effective and varied seasonally, as well as with wastewater temperature. PMID- 11424737 TI - Extraction and characterization of organic matter from surface waters (reservoir of Keddara in Algeria). AB - The combined XAD-8 and XAD-4 resin procedure for isolating dissolved organic solutes from Keddara reservoir in Algeria was found to isolate 65% of total organic carbon (TOC). Of the 45% of solute adsorbed onto XAD-8 resin, 34% was in the form of fulvic acids, and 11% humic acids. Approximately 19% of the hydrophilic solutes were adsorbed onto XAD-4 resin. Characterization experiments have shown that the three isolated fractions have a low apparent molecular weight investigated by ultrafiltration and fluorescence and low aromaticity as shown by the results of UV-absorbance, the formation potential of total organic halogens (reactivity with chlorine) and the polyhydroxyaromatic (PHA) determined by pyrolysis gas chromatography/mass spectrometry (GC/MS) which show that the isolated fractions are aliphatic as well as aromatic. The low molecular weight and the low aromaticity compared with literature data are due to coagulation and sedimentation of NOM with high molecular weight, which occur naturally in the reservoir. The Keddara reservoir, with low hydraulic loading area and long detention times receiving water with low NOM and high in hardness can be expected to be very effective coagulation and sedimentation basins. PMID- 11424738 TI - Tc-99m-leukocytes--is it an intracellular labelling? AB - Radiolabeled leukocytes have a potential for clinical use in detecting sites of inflammation. The increasing interest in the use of Tc-99m-labeled white blood cells (WBC) encourages exploration into the site(s) of binding of Tc-99m to the WBC components. Here we present the differential centrifugation study of the labeled leukocytes using a simple and low cost technique (SnTec). The results show most part of radioactivity bound to cytosol. We concluded that this is an intracellular labeling procedure that has the citoplasm, ribosomes unbound to membrane and soluble molecules as targets. PMID- 11424739 TI - Epitope expression in nine commercial kits for the determination of anti-thyroid peroxidase (TPO) antibodies. AB - Anti-thyroid peroxidase (TPO) antibodies, from patients with autoimmune disease, bind predominantly to two neighbouring, non-identical, conformational domains referred to as domains A and B. In recent years a number of ELISA assays have been developed for the detection of anti-TPO antibodies, however, considerable variation between the different commercial assay kits has been documented in inter-laboratory surveys (UK NEQAS). This investigation assessed the differences between nine commercial ELISA assays currently available in the UK. The anti-TPO kits varied in terms of their imprecision and accuracy and in the density of coated antigen. Recombinant antigen containing kits demonstrated partial destruction of the B epitope, possibly due to the close proximity of both epitope regions in the recombinant molecule. None of the kits expressed only one epitope although there were differences in the degrees of expression of each epitope. Clinicians should be aware of the variability of the numbers generated, when interpreting test results. PMID- 11424740 TI - Eosinophil activation in the tissue: synthetic steroid, budesonide, effectively inhibits the survival of eosinophils isolated from peripheral blood but not nasal polyp tissues. AB - We investigated the effect of a potent synthetic steroid, budesonide (BUD), on the survival of nasal polyp (NP) derived eosinophils (EOS). BUD, at the highest dose used, 10(-6) M, decreased this survival but only by approximately one third. We speculated that the relatively small inhibitory effect of budesonide on the survival of NP-EOS could be the result of these EOS being exposed to substantial amounts of GM-CSF, IL-5 or IL-3. In this regard, we detected 148 pg of GM-CSF per 150 mg of tissue, which approximately contained 106 of eosinophils, in the supernatant of NP explants for 24 h without any stimulation. Contents of both IL 5 and IL-3 were much less. We further studied survival of PB-EOS exposed to rhGM CSF and found that 10(-6) M of BUD could only inhibit by less than one third the survival of PB-EOS exposed to an amount of rhGM-CSF, similar to that detected in the supernatant of NP explants. In addition, we exposed PB-EOS to 200 pg/ml of rhGM-CSF for a relatively long period of time (4 days) in order to mimic chronic exposure in the tissue and found that the survival of these cells was prolonged to the extent similar to that observed in NP-EOS. Our data suggests that the prolonged spontaneous survival of NP-EOS ex vivo is likely the result of sustained in vivo exposure to GM-CSF and budesonide has a smaller inhibitory effect in the survival of these eosinophils as compared to those from peripheral blood. PMID- 11424741 TI - Natural antibody status in patients with Hashimoto's thyroiditis. AB - Raised levels of circulating natural antibodies (NABS) have been found in association with systemic lupus erythematosus (SLE) and chronic active hepatitis (CAH), indicative of polyclonal B-cell activation associated with these relatively non-organ specific autoimmune diseases. This study examined the natural antibody response in the organ-specific autoimmune disease Hashimoto's thyroiditis. Serum samples obtained from 69 women with newly diagnosed Hashimoto's thyroiditis together with 64 controls were analysed for IgG and IgM NABS directed at DNA, actin, myoglobin, myosin, trinitrophenyl hapten (TNP) and tubulin as the NAB antigen panel using an established ELISA. The same technique was also used to estimate thyroglobulin and thyroid microsomal autoantibody activities. Compared to a reference panel of normal serum samples, 31 of the Hashimoto's samples showed a greater than 2SD elevation of IgG and/or IgM NABS against one or more of the panel antigens together with elevated IgG thyroglobulin and thyroid microsomal antibody levels. The cases positive for one or more of the NAB panel also showed a greater incidence of active Hashimoto's thyroiditis as indicated by the presence of antibodies directed against the thyroid specific antigens. The above findings suggest that raised levels of NABS are also a feature of this organ-specific autoimmune disease. The wide ranging NAB specificities involved are consistent with an underlying or epiphenomenal state of polyclonal B-cell activation. PMID- 11424742 TI - [Facing an aggressive client--psychological consequences for employees]. AB - The main aim of the study was to assess the frequency of violence at work experienced by employees, representing selected occupational groups, and to identify forms of aggression manifested by clients. We also tried to find out whether the frequency of aggressive behaviours of clients differentiates the intensity of fear attitudes. In addition, we made an attempt to discover whether the intensity of fear attitude towards clients is associated with perceived social support and burnout. The study covered 248 employees: social workers of the Municipal Welfare Centre, as well as inspectors of the State Labour Inspectorate and the Revenue Office. The study revealed that social workers were mostly exposed to aggressive behaviour of clients, and they showed the most fearful attitude towards contacts with them. This attitude was concomitant with burnout. The results of the study suggest that the contact with aggressive behaviour encountered at work is conducive to inappropriate treatment of clients, job dissatisfaction and the feeling of senselessness. As indicated by the study, the conviction that one may count on the support superiors and colleagues suggest in stressful situations at work mitigate the fear attitude towards clients. PMID- 11424743 TI - [Analysis of morbidity and testing of trends for qualitatively variable incidence of occupational diseases induced by industrial dust in selected workplaces]. AB - The paper addresses the problem of occupational diseases induced by industrial dust (items 2-4 on the Polish list of occupational diseases) during the years 1970-89. The data were obtained from the Section of Work Hygiene of the former Sanitary and Epidemiological Station in Zielona Gora. In the population of workers under study, the incidence of chronic respiratory diseases (CRD) (items 3 and 4 on the Polish list of occupational diseases) was higher as compared with pneumoconiosis in terms of both the number of cases and the number of plants where they were observed. In case of pneumoconiosis, welding smoke and foundry dust were found to be etiologic factors responsible for its incidence, and organic dusts of plant and plastic origin together with industrial dust induced CRD. First cases of CRD were observed in the exposure interval of 6-10 years, and pneumoconiosis already in the interval of 0-5 years. The intensified incidence of both pneumoconiosis and CRD was observed after 11-25 years of occupational exposure. An upward trend in the incidence of pneumoconioses determined by the concentration of industrial dusts and the duration of exposure was found in one plant, and of CRD determined by the same factors in two plants under study. PMID- 11424744 TI - [Evaluation of late results for surgical treatment of chronic lateral epicondylitis]. AB - The results of surgical treatment of 25 patients with lateral epicondytilis (tennis elbow) were analysed. The mean time before surgery was 2 years and 4 months. The patients' negative attitude towards surgery and the prolonged conservative treatment by general practitioners were the reasons for such a long period of ineffective conservative procedures. Following Hohmann's surgery the flexion in elbow joint and dorsal flexion in radio-carpal joint limitation were the most common residual impairments. The loss of muscle circumference of the operated extremity was observed in 10 (40%) patients. Pain subsided in 20 (80%) and numbness in 11 (44%) patients. Two third of the patients were satisfied with the surgery outcome This moderately good general outcome was probably due to the prolonged time passed since the onset of symptoms and repeated local steroid injections. The majority of manual workers returned to their work. PMID- 11424745 TI - [Occupational monitoring of lung cancer in epidemiologic studies]. AB - This paper addresses the problem of diagnosing occupational lung cancer faced in different countries. The risks of lung cancer attributed to occupational exposure in selected epidemiological studies are presented. Evidenced and probable lung carcinogens, as well as occupations and industries responsible for the increased risk for the incidence of this pathology are discussed in the light of epidemiological studies. PMID- 11424746 TI - [Proposals of rational procedures for certifying occupational voice disorders]. AB - Following a long period of experience acquired in assessing occupation-related voice disorders, the proposals of objective measures, facilitating the certification of occupational disease of the voice organ in accordance with the binding legal regulations, are presented. The importance of documentary evidence of at least two years of treatment and phoniatric rehabilitation is stressed. A need to prevent occupation-related voice disorders through phoniatric diagnostic procedures is highlighted. PMID- 11424747 TI - [Rules for prevention and certification of occupational diseases in workers occupationally exposed to antineoplastic drugs]. AB - The risk of occupational exposure to antineoplastic drugs is related to cancerogenesis and reproductive dysfunction, including fertility disorders, congenital defects and toxic effects inter alia on the liver, kidneys, skin and bone marrow. The authors review current legal regulations of antineoplastic drug administration, the present state of the patients' care and guidelines for prophylactic examinations in this occupational group. PMID- 11424748 TI - [Genetic polymorphism of glutathione s-transferase as a factor predisposing to allergic dermatitis]. AB - In the inductive phase of contact allergic dermatitis, simple chemical compounds (haptens) produce together with epidermic proteins adducts presented by Langerhans cells to T lymphocytes. Binding to protein carrier is a necessary condition of transforming a low-molecular allergen into immunogenic one and evoking immunological reaction. The production of allergen adducts with proteins is conditioned by the presence of electrophilic groups in their molecules, or their acquiring during biotransformation phase I. Active allergen metabolites undergo further alterations during biotransformation phase II which leads most frequently to the decline in their chemical activity and more rapid excretion from the body. The number of reactive metabolites (reactive allergens) available for producing adducts with proteins keeps the balance between activation and deactivation reactions. Glutathione S-transferases play a particular role in the allergens (or their metabolites) deactivation process in biotransformation phase II. These enzymes catalyse reactions responsible for the declined electrophilic potential of allergens (or their metabolites), and thus for the decrease in the number of allergen molecules able to produce protein covalent bindings (adducts). Glutathione S-transferases, occurring in the human cellular cytoplasm belong to five classes: alpha(GST A), mu(GST M), theta(GST P), pi(GST T) and Z(GST Z), as well as to one class present in microsomes. The study indicated the presence of isoenzymes GST T1 and GST M1 in the skin. Both isoforms participate in the process of low-molecular allergen biotransformation. Carriers of defective genes GST T1 and/or GST M1 are more vulnerable to allergenic effect of some allergens, e.g. thimerosal, which is associated with the absence of or decrease in the activity of isoenzymes GST T1 and GST M1. PMID- 11424749 TI - [The effects of work in an air-conditioned cabin and stay in a resting chamber on blood pressure and heart rate of operators employed in deep copper mines]. AB - This paper is part of a wider comparative study of the heart rate, blood pressure, external and core temperature in operators of self-propelled mining machines with and without air-conditioning cabins. Two groups, each of ten operators, characterised by the similar age and duration of employment, stayed for 20 min a specially prepared resting chamber with much more advantageous microclimatic conditions. The results of our examinations (Holter heart rate and continuous blood pressure recordings, external and core temperature measurements) revealed that during the work (particularly during the increased work-load) all parameters recorded were significantly lower in air-conditioning cabins as compared with the group working without air-condition. In both groups, a complete restitution of the heart rate and blood pressure was observed after a 20-min stay in the resting chamber. During the work, a statistically significant increase in the external temperature was found in both groups of operators, whereas the increase in the core temperature was observed only in operators working without air-condition. After a 20-min stay in the resting chamber, a complete return to the normal temperature was noted only in operators working in air-conditioned cabins. PMID- 11424750 TI - Conflict of interest and the rebuttable presumption in the context of an excess benefit transaction. PMID- 11424751 TI - Court denies relief to excluded emergency physicians. PMID- 11424752 TI - Survey provides a snapshot of staffing challenges in the OR. PMID- 11424753 TI - Interviews find some ORs have vacancies, others waiting lists. PMID- 11424754 TI - Temp staff become a fixture in ORs. PMID- 11424755 TI - Staff's plan attacks OR vacancy rate. PMID- 11424756 TI - A bridge between school and service. PMID- 11424757 TI - Video opens OR to a host of questions. PMID- 11424758 TI - Tips on managing pass-thru payments. PMID- 11424759 TI - Be proactive on advance directives. PMID- 11424760 TI - How to hire the best for your ASC. PMID- 11424761 TI - "Digital hospital" planned in south. PMID- 11424762 TI - [Perinatal and newborn care in the Emilia-Romagna Region health planning]. AB - The report summarizes the contents of a conclusion of a document of the workshop "Neonatal and Prenatal Intensive Care" activated by the Emilia-Romagna Region to set up the third Regional Health Planning 2000-2002. The proposals, wholly taken from the Regional Program, aim to improve the care improving the outcomes in terms of mortality and maternal-infant disability. The organizing model proposed is that of integrated networks of services distributed according to a hierarchy of functions (Hub & Spoke model): 1st, 2nd, and 3rd level of functions. The network services should function as an integrated system, governed by agreed clinical and organizational guidelines, in which requirements of equipment sets and professional abilities are identified, as well as, relationship criteria among the different levels that regulate the by-directional course of the patients and/or the operators: the continuous training activity, the monitoring of a set of process indicators and results and also the clinical-organizational audit activity, are the instruments identified for results verification and for continuous actions of improvements. PMID- 11424763 TI - Current obstetrics: what neonatologists need to know. AB - The changes in obstetrics have led to the collection of much more physiologic data about the fetus during development. These advances are not only important to obstetricians but also to neonatologists. Unfortunately, the organizational structure of medicine has made it difficult to develop effective perinatal medical services, where both a full complement of obstetrical resources and a full complement of neonatal services are available within the same hospital service. The reasons for this are discussed. Neonatologists should use the current fetal physiologic data which are collected to develop both short and long term follow up studies in order to relate specific obstetrical findings to neonatal outcome. The advances in obstetrics have had a particular impact upon the problem of intrauterine growth retardation. This has meant that neonatologists must deal with infants born very premature who are also intrauterine growth retarded. These babies may have obstetrical data defining fetal circulatory failure and/or placental failure. It is in this arena where a knowledge of how to interpret the physiologic data collected by obstetricians about the fetus is particularly useful to neonatologists. PMID- 11424764 TI - [Amnioinfusion: techniques, indications, and controlled retrospective study of 55 cases]. AB - Amnioinfusion is a relatively recent procedure introduced among fetal medicine techniques. Its applications focus on two different methods: transcervical and transabdominal. The first procedure usually is carried out during "intrapartum amnioinfusion" to prevent or treat fetal heart rate (FHR) decelerations related to oligohydramnios or to dilute thick meconium staining of the amniotic fluid. The latter method used during "antepartum amnioinfusion" is usually indicated for severe oligohydramnios in order to avoid the complications related such as pulmonary hypoplasia, deforming effects of oligohydramnios, variable FHR decelerations and intraventricular hemorrhages. Antepartum amnioinfusion, also used to improve ultrasound visualisation in presence of oligohydramnios, is less employed as compared to intrapartum amnioinfusion, therefore its risks are not well established. In order to study possible adverse effects on the mother or foetus, fifty five patients affected by oligohydramnios at 17th-34th week of gestational age were submitted to antepartum amnioinfusion (1-5 procedures) and were matched retrospectively with forty seven women with the same characteristics treated with the conservative and expectant management. The trend of pregnancy was the same for both groups in relation to maternal fever > 38 degrees (10.9% in the amnioinfused group vs 17.0% in control group ns), leukocyte count > 18,000/mm3 (25.5% vs 21.3%, ns), C-reactive protein > 10 ng/ml (10.9% vs 6.4%, ns). The latency period between admission and delivery was significantly longer in the amnioinfused group than in the control one [21 (range 1-98) vs 9 days (range 0-72); p < 0.001] and the frequency of Apgar score < 7 at the 5th min was less represented in the amnioinfused group than in the control group (32.3% vs 66.6%; p < 0.001). In conclusion, it was interesting to note that antepartum amnioinfusion seems to increase the latency period between premature rupture of membranes and delivery, but it remains to clarify if this procedure is as much safe for the fetus as for the mother. PMID- 11424765 TI - [Comparative analysis of premature labor markers]. AB - We compared the diagnostic value of foetal fibronectin (fFN), phosphorilated insulin-like growth factor protein binding-1 (Birth test) and insulin-like growth factor protein binding-1 (PROM test) as markers of premature rupture of membrane (PROM) and in prediction of preterm labor with intact membranes. The study population included 120 asymptomatic women (group 1), with gestation of 24-34 weeks; we also considered 21 patients with clinically confirmed PROM (group 2) and 26 patients with suspected PROM with gestation between 15 and 24 weeks (group 3) (as measured by sonography data). From our data, it seems that only the predictive value of each test is related to the characteristics of the population considered. The fFN test and the Birth test prove to be highly predictive in pregnant women without PROM, on the contrary the use of the PROM test is optimal in pregnant women with suspected PROM or with PROM, regardless of risk factor or with contractile activity. PMID- 11424766 TI - [Pre-partum cardiotocography: comparison between the traditional and the computed systems]. AB - The objective of our study was to determine the usefulness of a computerized antepartum fetal heart rate (FHR) analysis instead of the common visual evaluation of cardiotocographic parameters. From a total of 4,000 antepartum FHR tracings performed in our institute in the years 1994-1996, and analyzed on-line by the software HP 2CTG, 100 were visually assessed by five physicians in three different moments. To each reader a qualitative and quantitative interpretation was required. With use of k coefficient, we have analyzed the differences in observer's qualitative interpretation, then, with use of the intraclass correlation coefficient (RINTRA) and the contingency table X2 we have compared the quantitative analysis with that of the computer. Our results show that variability is the most difficult parameter to interpret, so that the interobserver agreement isn't in anyway sufficient (k 0.43). The agreement between observers on the clinical judgment of the FHR traces proves to be low (k 0.54). The agreement between each of the observers and the computer readings is good for the estimation of the FHR (RINTRA between 0.96 e 0.98) and low for the mean term variability (RINTRA between 0.81 and 0.58). There is a valuable agreement with the computer for tracings with 0 or 1 large acceleration (94.7% and 97.5% respectively), but it is significantly poorer (p < 0.001) for tracings with 0 or 1 small acceleration (57.4% and 50.8% respectively). Decelerations are the most difficult pattern of interpretation. Usually observers do not identify 36% of the FHR traces having at least one deceleration according to computer analysis. Our study confirms the wide variability in fetal monitoring interpretation, a difficulty which can be overcome thanks to a computerized analysis. PMID- 11424767 TI - [Fetal growth restriction and insulin-like growth factors]. AB - The insulin-like growth factors (IGFs) and their receptors are implicated in pre- and postnatal growth and development. It is believed that the alteration in their activity may contribute to intrauterine growth restriction (IUGR). The aim of this experimental study was to relate some metabolic alterations, involving insulin-like growth factor pathway, in the placenta of pregnancies complicated by intrauterine growth restriction. Placental samples were obtained from six uncomplicated pregnancies and four pregnancies complicated by IUGR. These samples were then stained by immunohistochemical technique, using monoclonal antibodies. Our data have not shown a significant difference in the IR, the Shc isoforms and Akt levels between normal and IUGR placentas. The IUGR placentas had significantly lower levels of IRS-2 expression and higher levels of p85 transcription. IGF-I receptor binds to its ligand and activates two intracellular processes mainly a Shc-mediated pro-mitotic pathway and an anti-apoptotic pathway mediated by IRS and Akt. The diminished activity of one of the two pathways may alter the mitosis/apoptosis balance. Because of the low number of samples and the knowledge about the enzymatic pathways, we have not been able to associate our data to any biological consequence. We can only demonstrate that the enzymatic differences between IUGR and controls tend to the pro-apoptotic processes. It seems to be an other in vivo aspect in favour of the placental role in the IUGR pathogenesis. PMID- 11424768 TI - [The relative weight of Doppler of the uterine artery and 24-h ambulatory blood pressure monitoring in predicting hypertension in pregnancy and preeclampsia]. AB - OBJECTIVE: To assess the combined use and the "relative weight" of Uterine Artery Doppler Velocimetry and 24-h Ambulatory Blood Pressure Monitoring (ABPM) to predict Gestational Hypertension (GH) and Preeclampsia (PE). METHODS: The predictivity of Doppler of uterine artery flow at 24 weeks and of ABPM at 24-30 weeks was assessed in 75 normotensive primigravidae, considering the subsequent onset of GH and/or PE. RESULTS: In our series the sensitivity of Doppler versus GH is 62%, and 57% vs PE. The altered ABPM shows a sensitivity of 97% vs GH and 99% vs PE. In the cases with the both tests positive it was 97% vs GH, and 99% vs PE. In the subgroup of patients with altered Doppler velocimetry and normal ABPM, the sensitivity vs GH and PE is 12% (PPV = 12%). A second subgroup with normal Doppler velocimetry and altered ABPM shows a sensitivity of 94% vs GH (PPV = 88%), and 99% vs PE (PPV = 33%). CONCLUSIONS: Uterine Artery Doppler Velocimetry and 24-h Ambulatory Blood Pressure Monitoring (ABPM) have a good predictive power for GH and PE, but the combined use improves the efficacy to identify an abnormal outcome of pregnancy. When evaluating for relative weight, the ABPM is more powerful in predicting for GH and PE than Doppler assessment. However, the high specificity of Doppler velocimetry reduces the false-positive ratio in the combined screening during the 2nd trimester of pregnancy. PMID- 11424769 TI - [Effects of betamethasone administration on maternal-fetal Doppler velocimetry]. AB - BACKGROUND: The effect of antenatal betamethasone on fetal parameters includes a transient reduction of fetal heart rate variation and of fetal body movements. An effect on maternal-fetal blood flow has also been shown, with non-univocal results. AIMS: To evaluate the effect of antenatal betamethasone in third trimester singleton high-risk pregnancies by Doppler technology. SUBJECTS AND METHODS: Thirty-six pregnant women who received a full course of betamethasone (12 mg i.m. two times, 24 hrs apart) were studied. The Doppler examination included the assessment of the pulsatility index (PI) of the umbilical artery (UA PI), the middle cerebral artery (MCA PI) and of resistance index of uterine arteries (Ut RI) before treatment, and 48 and 96 hrs after second dose of betamethasone. RESULTS: No significant variation was noted in UA PI through betamethasone therapy. MCA PI decreased significantly 48 hours from the last injection of betamethasone (p = 0.004), and returned to basal values at 96 hours. No difference was found for the other Doppler parameters examined. When serial Doppler studies were analyzed according to the gestational age, in the group < 32 wks' gestation, MCA PI decreased significantly after 48 hours (p < 0.006) and returned to pre-treatment values after 96 hours from the last betamethasone dose. Conversely, no difference was found in Doppler serial measurements in any of the analyzed districts in the subgroup > or = 32 wks' gestation. CONCLUSION: Betamethasone treatment is associated with an increase in end diastolic flow of the middle cerebral artery, as reflected by a significant, although transient, reduction of MCA PI, especially at gestational ages < 32 weeks. PMID- 11424770 TI - [Perinatal morbidity and mortality in children born to mothers with gestational hypertension]. AB - Gestational hypertension is a severe pathology leading to important maternal and neonatal effects. It represents one of the most important causes of maternal morbidity and contributes to a high percentage of perinatal mortality, determined by fetal hypoxia and especially by prematurity and low birth weight. To-date the only treatment offered in gestational hypertension remains delivery, which has to be scheduled regarding timing and method on the basis of the appearance of hypertension, of its severeness and maternal and fetal complications. To evaluate the clinical course and the presence of hypertensive risk factors, a clinical epidemiological study was carried out on two groups of pregnant women: a group made up of 50 women with hypertension and a control group of 80 women in whom no gravidic pathology arose. We hence were able to reveal the risk factors associated with hypertension in pregnancy such as maternal age, nulliparity, and elevated body mass index. Regarding neonatal prognosis, we observed a higher incidence in premature birth (30th- to 38th week of gestational age) and the need for elective or urgent caesarean sections, with respect to the spontaneous deliveries observed in the control group. We also observed reduced weight of both placenta and the newborn in hypertensive mothers with significant statistical differences between the two groups (p < 0.001). Evaluation of neonatal data at birth showed lower Apgar indices at 1st and 5th minute from birth in the study group with a higher percentage of newborns transferred to the neonatal intensive care unit. We also observed a high percentage of still-births equal to 14% in the study group as opposed to the control group. PMID- 11424771 TI - [Combination of the computerized analysis of fetal heart rate and amniotic fluid index in the prediction of neonatal acidemia: a modified biophysical profile]. AB - OBJECTIVE: To study the use of a modification of the classical biophysical profile by the combination of computerized analysis of fetal heart rate (cCTG) and the amniotic fluid index (AFI) in the prediction of neonatal acidemia at birth. METHODS: We considered 89 singleton third trimester high risk pregnancies delivered by cesarean section, with an AFI evaluated within 24 hrs from birth, and an antepartum cCTG performed within 6 hrs from delivery. We assigned values for AFI (oligo/anhydramnios = 1, normal = 0) and cCTG (Dawes-Redman criteria, not satisfied = 1, satisfied = 0). The endpoint was to predict an abnormal neonatal outcome as defined by an umbilical artery (UA) pH < or = 7.2. RESULTS: Fifteen neonates presented an UA pH < or = 7.2. By performing a logistic regression, we found that cCTG + AFI score (abnormal value 1-2) was able to significantly predict a pH value (< or = 7.20) with an Odds Ratio = 2.83 (p < 0.02). The diagnostic accuracy of the combination of cCTG + AFI was as follows: sensitivity: 80%, specificity: 58%. COMMENT: We suggest that the combination of cCTG + AFI included in the simple score we propose, may be of value in the prediction of neonatal acidemia and help in the management of third trimester high risk pregnancies. PMID- 11424772 TI - [Pulsatility index of the umbilical artery and fetal movements/hour in the prediction of neonatal acidemia in high risk pregnancy]. AB - OBJECTIVE: The aim of this study was to correlate antepartum computerized cardiotocography (cCTG) and Doppler velocimetry parameters to umbilical blood gas analysis (UBGA) value and Apgar score 1-5 min as neonatal outcome endpoints. METHODS: Forty-eight third trimester single high risk pregnancies were considered for this study, with a cCTG performed within 24 hrs from delivery, a complete Doppler velocimetry study performed within 48 hrs from delivery and an UBGA evaluation at birth and before the first breath. RESULTS: When we analyzed the backward stepwise regression of each Doppler velocimetry and cCTG parameters versus UBGA parameters as dependent variables, we found that the linear combination of fetal heart rate (FHR) (p < 0.001), high variation episodes in min (HV) (p < 0.01) and low variation episodes in msec (LV) (p < 0.03) was able to predict pO2 values. When performing a logistic regression of data for every single parameter of FHR tracing and Doppler velocimetry against pH < 7.2 and Becf < -4 as endpoints, we found that only umbilical artery pulsatility index (UA PI) was able to predict umbilical artery pH (O.R.: 8.1 [1.07-61.8]) and only fetal movements (FM) from FHR tracing values was able to predict UA pH (O.R.: 0.94 [0.89-0.99]). Further analysis considers a cut-off for the prediction of UA pH < 7.2 at birth a value at > or = 1.35 for UA PI and at < or = 12 for FM/h. The combination of UA PI > 1.35 and FM/h < 12 did not improve the ability to predict acidemia at birth. CONCLUSION: The validity of our data from the clinical point of view suggests that in third trimester high risk pregnancies, an UA PI > or = 1.35 and/or FM/h < or = 12 (in a FHR tracing > or = 40 min) may represent a risk of 70% acidemia in neonates. PMID- 11424773 TI - [Echographic measurement of subcutaneous adipose tissue as fetal growth index]. AB - OBJECTIVE: To evaluate, by 3rd trimester ultrasound examination, fetal adipose tissue to predict fetal growth and birthweight. PATIENTS AND METHODS: Thirteen pregnant women (group 1) with uncomplicated pregnancy and 9 diabetic women (group 2) have been enrolled. We have examined adipose tissue at the cross sectional level of arm, thigh and abdomen and at the longitudinal sectional level of shoulder. During 3rd trimester measurements were taken at the beginning (28-32 gestational weeks), at the middle (34-38 gestational weeks) and at term (39-41 gestational weeks). At birth we recorded the skinfolds of arm, thigh and shoulder and the neonatal weight. Subsequently, by regression analyses ultrasound parameters of physiological pregnancies were correlated to skinfolds measurements at birth, while abdomen thickness was compared to birthweight. In addition, ultrasound markers of the physiological pregnancies were compared to those of diabetic pregnancies by the t-test. RESULTS: Fetal limbs examination had not a prognostic value at any gestational week. Shoulder ultrasound measurements appeared well correlated to the shoulder skinfold after birth since the 34th week. Diabetic mothers' fetuses presented much more adipose tissue than control cases, particularly of arm and thigh at 28-32 weeks, of shoulder at 34-38 weeks and in all the markers at term pregnancy. CONCLUSION: Ultrasound examination of fetal adipose tissue seems to have a predictive value for fetal growth both in physiological and diabetic pregnancy. PMID- 11424774 TI - [Computerized analysis of antepartum fetal heart rate and maternal glycemic levels in pregnancy complicated with insulin-dependent diabetes]. AB - We aimed to evaluate the effect of maternal glycemia on the parameters of CTG as assessed by a computerized fetal heart rate interpretation system (cCTG). We studied prospectively 22 single insuline dependent diabetic pregnancies (34-39 wks' gestation). A negative correlation was found between high variability epochs (HV) in msec, and maternal glycemia (r = -0.58, p < 0.01). HV epochs are known to be related with hypoxic episodes in the fetus. We suggest that the evaluation of maternal glycemia at the time of cCTG may increase the diagnostic accuracy of automated fetal heart rate interpretation. PMID- 11424775 TI - [Incidence of thyroid diseases in pregnant women with type I diabetes mellitus]. AB - The interaction between pregnancy, Diabetes Mellitus and thyroid disturbance needs a particular attention. The antithyroid antibodies are more frequent in pregnant women with insulin-dependent diabetes mellitus (IDDM) than in normal pregnant women. Beside, an increased prevalence of subclinic ipotyroidism has been described in pregnant diabetic women. The object of this study is to verify if women with insulin-dependent diabetes mellitus have more probability than normal women to develop a thyroid pathology in pregnancy, in an area with sufficient iodine. Twenty-eight women have been studied: 15 with IDDM, with age 20-37 years, and 13 healthy women, with the same age. Patients were evaluated at the following time-intervals: 9-12 and 18-20 weeks' gestation, at delivery and six months after delivery. Diabetic women followed an insulinic therapy optimized for to reach a good metabolic control. All follow the patients had a thyroid ecography to evaluate thyroid volume and possible presence of nodular formation or others pathologic signs. A blood sample to assay FT3, FT4, TSH, of the antibodies Anti-Thyreoperoxidase (Anti-TPO) and Anti-Thyreoglobulin (Anti-TG). A urine specimen taken to evaluate the iodine excretion. No significant difference was observed between diabetic and normal women, for the values of TSH (p < 0.2), FT4(p < 0.7), FT3(p < 0.6). Instead a significant difference was found between the thyroid volume (p < 0.04), in the diabetic patients versus the normal women, at delivery and six months after delivery. The results of this study underline the importance of the screening of the thyroid function and morphology, in all the pregnant women and, particularly, in the diabetic patients, to find the presence of glandular alterations as early as possible. PMID- 11424776 TI - [Neonatal anthropometry and early differentiations in embryo-fetal growth]. AB - It is well known that the biologic variability in fetal size increases as pregnancy advances, although the embryonal and early fetal growth patterns as well as how early and how much the genetic, hormonal and environmental variables play a role in its modulation are still debated. It is accepted that growth in the first trimester of pregnancy is relatively uniform, with a minimal biologic variability; this variability may be underestimated, because the transversal studies do not permit the identification of the growth pattern. The aim of this work is to evaluate, by means of a longitudinal study, the time of embryo-fetal growth differentiation related at neonatal anthropometric measurements. We evaluated 238 neonates (123 female; 115 male) delivered at term after low risk, uncomplicated pregnancies. The subjects were divided into three tertles (low, mid and high) according to birth weight, length and head circumference. For each tertle, distance curves, velocity curves, and rate of increase were calculated by using respectively fetal abdominal circumference (for birth weight), fetal femural length (for neonatal length) and fetal head circumference (for neonatal circumference). The distance curves showed clear differences among the tertles only in the second period of pregnancy, whereas the velocity curves showed clear differences among tertles already in the first 12 weeks. The value of growth rates were similar for all the variables during the entire time considered. This study shows that the anthropometric differences between newborn subgroups exist already at the end of the first trimester of pregnancy and, in physiological conditions, until the end of pregnancy. The anthropometric differences observed early in our study, at twelve weeks of gestational age, are still present at the end of pregnancy and let us suppose a very early expression of the genetic potential for individual growth. PMID- 11424777 TI - [Evaluation of the peripartum effects of 2 analgesics: meperidine and tramadol, used in labor]. AB - The need for analgesia to overcome pain in labour is highly requested by women today. Various ways either non pharmachologic e.g. Emotional sustain, psycho prophylactic preparation, yoga and hypnosis or pharmachologic such as epidural blockade or parenteral are used. Therefore in our study we evaluated the efficacy and tolerability of the two opioids usually used today in parenteral analgesia to reduce pain during labour: Tramadol and Meperidine. We studied two groups of patients each made up of 20 women in labour, all at term and with a physiologic course of pregnancy. 75 mg i.m. of Meperidine chloryhydrate were somministered in the first group while in the second group 100 mg i.m. of tramadol chloryhydrate were somministered. Various maternal, fetal and neonatal parameters were then monitored demonstrating--A moderate maternal analgesic effect in both drugs (evaluated through the analogic grading of pain). In the group to whom Meperidine was given, sedative effects on the mother were observed associated with respiratory depression in the newborn (the latter evaluated through the Apgar index at 1st and 5th minute of life and pH of the blood obtained at the umbilical cord. The data obtained permitted us to conclude that Tramadol in accordance to the obtained in literature gives an analogous analgesic effect, with better tolerability for the absence of collateral effects on the mother, fetus and newborn. PMID- 11424778 TI - [Placental histopathology and perinatal surveillance]. AB - The histological examination of the placenta helps to better understand both pathological pregnancies and fetal growth defects, therefore the true beneficiary of the histological diagnosis is the newborn. Of the 492 newborns born between the 1o April and 30 June 2000 we analyzed 142 placentas, following a modified Macpherson e Szulman's protocol. Only 73 of the 142 newborns have been hospitalized in the pathological division. The placental histological diagnosis of the remaining 69 hospitalized in "Rooming in" showed that only 10 placentas were in the normality; 22 placentas showed alterations referable to a dysmetabolic, dysglicicemyc and hypertension maternal pathology without there being lesions that could interfere with the development and maturation of the fetus; 37 placentas showed indicative lesions of hypoxic and ischemic fetal sofference of varing gravity. These data show how the histological examination of the placenta has many benefits in the immediate care of newborns, in subsequent evaluations and in the pediatric age. PMID- 11424779 TI - [Leonardo da Vinci and his studies on the human fetus and the placenta]. AB - OBJECTIVE: To review the accuracy of Leonardo's anatomical studies of the female external genitalia and the foetus, particularly those concerning the umbilical cord with its blood vessels. BACKGROUND: The anatomical drawings of Leonardo da Vinci which are now stored in the Windors Castle near London were reviewed and the accuracy of the details of the genital apparatus and foetus was evaluated. A written comment characterizes many of his drawings. RESULTS: He described accurately the position of the uterine blood vessels and the relationship between the pelvic organs. However his drawing and description of the female external genitalia and human placenta was incorrect because his understanding of it was inadequate. He believed that the human placenta had cotyledons like that of the ungulate uterus (drown side A of sheet 19). At the top of that sheet some Leonardo's details of the anatomic relationship are shown. CONCLUSIONS: In our opinion, Leonardo's misunderstanding is probably because he was only able to perform one anatomical dissection of a pregnant women and foetus and therefore his knowledge was limited. The resulting documentation of the pregnant women and foetus was therefore heavily supplemented with his enormous experience in animal anatomy. PMID- 11424780 TI - Prospects for a new synthetic surfactant. AB - Physiologically active artificial surfactants can be made from lipids and analogs of SP-C and SP-B, the latter serving as cross-linking peptides promoting the formation of a surface-associated surfactant reservoir. Artificial surfactants can probably be tailored to maximize resistance to inactivation by meconium and plasma proteins by modifying the amino acid sequence of the protein analogs or adding non-ionic polymers to the final product. Comparative clinical trials are required to evaluate the efficacy of these new artificial surfactants in various forms of neonatal lung disease. PMID- 11424781 TI - [Pre- and post-natal corticosteroids: side effects]. AB - It is well known that prenatal steroid therapy (ST) reduces the mortality rate and the incidence of respiratory distress syndrome and intraventricular haemorrhage in premature infants. The benefits and safety of repeated courses of antenatal ST are doubtful due to possible side effects in the mother and baby. Experimental studies in animals have shown that multiple courses of antenatal ST have deleterious effects on lung growth and organisation, brain myelination, hypothalamic-pituitary-adrenal function and retina development. In humans, exposure to multiple courses of antenatal ST is associated with small head circumference at birth and increased incidence of maternal endometritis and early onset neonatal sepsis. When administered soon after birth, ST may enable a reduction in ventilator settings and facilitate weaning from mechanical ventilation, but although associated with rapid improvement in lung function, it does not modify mortality or long-term outcome and has many acute side effects. Deleterious effects on lung maturation and neuro-developmental outcome, including cerebral palsy, have also been reported. The paucity of follow-up data is a major problem, and further prospective trials are needed. PMID- 11424782 TI - Timing of exogenous surfactant administration in clinical practice. Report of a multi-center Italian Observational Study. AB - We now have convincing data that surfactant administration in newborns with the respiratory distress syndrome (RDS) is effective in reducing the need for respiratory supports and improves the clinical outcomes. To evaluate the different strategies adopted in clinical practice for the surfactant use we performed an observational multicenter study. All newborns with gestational age (g.a.) between 24-32 wk. with RDS and that were in need of mechanical ventilation and treated with exogenous surfactant from 1994 to 1996 in twelve Neonatal Intensive Care Units have been evaluated. The patients have been divided into two groups: one includes babies that were given surfactant within the first 15 minutes of life (very early -VE- treated) and the other includes those that received the treatment at age 16- 180 minutes (early -E- treated). The analysis concerned 246 cases, 54 VE treated and 192 E treated. The two groups were comparable with regards to sex, Apgar score at one and five minutes, maternal treatment with steroids and multiple pregnancy. Mean birth weight and g.a. were significantly lower and the rate of caesarean sections was higher in the VE than in the E. Almost 80% of the VE newborns had a g.a. equal or less than 28 wks. One single dose of surfactant was administered in 77.8% of VE babies and 45.3% in the E group, while 18.5% VE and 36.8% E received two doses, 3.7% VE and 16.3% E were given three doses. The mean number of doses used per patient was 1.3 in VE and 1.7 in E. 62.5% of cases with g.a. equal to or less than 26 wks and 80.8% of those with g.a. 27-28 wk. received a single dose if VE, compared to 31.9% and 49.1% respectively if E treated. Prenatal prophylaxis of RDS with steroids, considering only the mothers that received a complete course between 48 hrs and 7 days before the delivery, did not modify the number of doses of surfactant required in both groups. The main outcomes did not differ in the two groups: mortality at 28 days of life, age at death, pneumothorax, intraventricular haemorrhage grade 3-4, retinopathy of prematurity grade 3-4, patent ductus arteriosus and oxygen dependency at 36 wks post-conceptional age. CONCLUSIONS: Very early surfactant administration in clinical practice reserved for newborns with the lowest g.a., improves the outcome and does not increase surfactant consumption. PMID- 11424783 TI - [Fetal lung maturity and fetal flowmetry changes]. AB - OBJECTIVE: To verify whether in high risk pregnancies, fetuses with altered Doppler flow velocimetry waveforms (FVW) are associated with both biochemical and/or biophysical changes in the amniotic fluid (AF) lung surfactant. MATERIALS AND METHODS: Hundred-eight high risk pregnancies were considered on the basis of a complete Doppler velocimetry study performed within 24 hrs before amniocentesis and delivery within 1 week from amniocentesis. From this group, 22 pregnancies were recruited as altered FVW cases on the basis of simultaneous alteration of Umbilical Artery Pulsatility Index (UA PI), and UA PI/MCA PI (Middle Cerebral Artery Pulsatility Index) ratio (UA/MCA); in all altered FVW group cases the Mean Uterine Artery Resistance Index (Ut RI) was also altered. Amniotic fluid samples obtained by ultrasound guided amniocentesis were analysed for FLM within 24 hours from collection. Pulmonary maturity was determined by planimetric lecithin/sphingomyelin ratio (L/S), phosphatidylglycerol (PG) and the lamellar bodies count (LBs). RESULTS: When FLM parameters were regressed versus FVW in the whole study group, we found a significant negative correlation between UA/MCA and L/S (r = -0.19, p < 0.05). No significant correlations were found between FVW indices and LBs. When we compared FLM parameters for gestational age subgroups (23-32 wks, > 33 wks), L/S values were significantly higher in patients with normal FVW indices in comparison to pathologic FVW pregnancies for gestational ages > 33 wks. This was reflected by a negative correlation between FVW and FLM (UA/MCA vs. L/S r = -0.54, p < 0.02; UA/MCA vs. PG, r = -0.60, p < 0.01). COMMENT: Our data suggest that fetuses with an UA/MCA ratio > 95 degrees are not protected from neonatal RDS, as usually felt, but show a delay in FLM, particularly in pregnancies > 33 wks. PMID- 11424784 TI - [Fetal lung maturity in biamniotic twin pregnancy]. AB - OBJECTIVE: To assess the need to perform amniocentesis for the evaluation of fetal lung maturity (FLM) in both amniotic sacs of twin pregnancies. STUDY DESIGN: A retrospective study. We considered 33 twin biamniotic pregnancies, delivering within 1 week from amniocentesis. FLM testing was performed by planimetric lecithin/sphingomyelin ratio (L/S), and lamellar bodies count (LB) according to our methodology. RESULTS: A strong correlation was found between L/S (p < 0.001, r = 0.59) and LB (p < 0.001, r = 0.69) values between smaller and larger twins. No significant regression was found between differences of L/S and LB among both amniotic sacs against gestational age. When stratifying FLM values < 34 and > 34 wks' gestation, again no difference was found in mean difference of L/S and LB before and after 34 wks. CONCLUSIONS: FLM should not be assessed in both amniotic sacs at any gestational age independent of weight and/or sex of the conceptus. PMID- 11424785 TI - [Prenatal steroids, cesarean section, and cerebral hemorrhage in newborns with birth weight under 1500 g]. AB - OBJECTIVE: To identify perinatal risk and protective factors for peri intraventricular hemorrhage (PIVH) in 214 very low birth weight (VLBW) infants. METHOD: Perinatal risk and protective factors for PIVH were examined by univariate analysis and by multivariate logistic regression analysis that adjusted simultaneously for the effects of year of birth and perinatal characteristics. Because mode of delivery and antenatal use of steroid represent the only factors, among those considered, that could in most cases be controlled by the clinician, we calculated for each of the four combinations the relative risk of PIVH. RESULTS: By univariate analysis, antenatal steroid administration, cesarean section delivery, increasing birth weight and increasing gestational age were protective perinatal factors. By using logistic regression, significant protective factors against PIVH included a complete course of antenatal steroid therapy and increasing gestational age. Intrauterine growth retardation was associated with an increased risk of PIVH. The relative risk of intraventricular hemorrhage for each of the four combination of mode of delivery and antenatal steroid exposure demonstrate that antenatal steroid exposure and cesarean section delivery are most protective with a relative risk of 14% compared with vaginal delivery and no antenatal steroid exposure. CONCLUSIONS: Association of antenatal steroid administration with cesarean section delivery has an important role in lowering the risk of PIVH in VLBW infants. PMID- 11424786 TI - [Nasal CPAP in newborns with birth weight under 1500 g]. AB - OBJECTIVE: To evaluate the efficacy and safety of early nasal continuous positive airway pressure (CPAP) in the pulmonary management of very low birth weight (VLBW) infants. METHOD: Since 1993 in our neonatal intensive care unit we decided to reduce the rate of tracheal intubation and mechanical ventilation (IPPV) using more extensively nasal CPAP. By Student's t test and c2 analysis, VLBW infants with gestational age < 32 weeks born in 1988-92 were compared with those born in 1993-97. RESULTS: The 144 enrolled infants were comparable for birth weight and gestational age. There was a difference in the rate of antenatal steroid administration and cesarean section delivery, significantly more used in the second period. In both groups, 93% of the infants required respiratory support. The use of IPPV decreased from 68% in the first period to 30% in the second, conversely the use of CPAP increased from 25 to 63%. With regard to the primary outcome, in the second period the mortality rate and the incidence of pneumothorax were significantly lower. CONCLUSIONS: As the long-term survival of VLBW neonates continues to improve, pulmonary management is increasingly directed at minimizing the invasivity. This study demonstrated that early nasal CPAP may be successfully used in most VLBW infants. Also in the smallest neonates the procedure is safe and effective and may be important in lowering pulmonary morbidity and subsequent mortality. PMID- 11424787 TI - [The "guaranteed volume" in pressure support ventilation reduces the risk of barotrauma in premature children with severe respiratory syndrome]. AB - Chronic Lung Disease (CLD) remains a major complication of prematurity, and barovolutrauma is thought to be one of the most important predisposing factors. In view of reducing the damage due to variability of tidal volume (Vt), we studied the application of the new option of volume guarantee (VG). We analyzed ventilatory data of 25 preterm infants with respiratory distress syndrome (RDS) in two situations: pressure support ventilation (PSV) + VG vs PSV alone. Data collected showed application of significantly lower peak inspiratory pressure (p < 0.001) and mean airway pressure (p < 0.001) with the constant Vt. We conclude that this ventilatory strategy, by reducing the barotrauma, could be considered a protective ventilation. PMID- 11424789 TI - [Ethical problems in congenital malformations]. PMID- 11424788 TI - [Bronchoalveolar lavage with diluted porcine surfactant for alveolar debris removal in newborns treated with mechanical ventilation]. AB - Lung debris in respiratory distress syndrome (RDS) and meconium aspiration syndrome (MAS) contribute to deteriorate pulmonary function. Surfactant lavage, also with minimal quantity of diluted surfactant, is an effective method for treatment of severe MAS and seems to be useful also in course of RDS evolving to chronic lung disease (CLD), by improving lung mechanics. Authors report a clinical study in which tracheobronchial lavage with surfactant (15 ml/Kg of diluted porcine surfactant) improved significantly lung function in 3 patients with RDS in prolonged mechanical ventilation and in 2 patients with MAS. PMID- 11424790 TI - [Transplacental teratogenesis]. AB - Exposure during pregnancy to drugs or environmental chemicals (named xenobiotics) may cause birth defects in the embryo. Little is known about the biochemical and molecular mechanisms of teratological susceptibility. Many embryotoxic xenobiotics are proteratogens when bioactivated by enzymes, such as cytochromes, peroxidases and prostaglandin synthase to reactive intermediary metabolites. These intermediates are free radicals or electrophiles, that oxidize or bind irreversibly to cellular macromolecules such as DNA, proteins and lipids, altering cellular function. Teratological susceptibility is determined by a balance among maternal xenobiotic elimination, embryonic bioactivation and detoxification of reactive intermediates, cytoprotection and repair of molecular lesions. The embryo is relatively susceptible to reactive intermediates due to immature pathways for the detoxification, cytoprotection and repair. In this way, teratogenesis can occur from exposure to therapeutic concentrations of drugs or low concentrations of environmental chemicals. PMID- 11424791 TI - [Incidence of chromosomal anomalies and congenital malformations in children born from assisted fertilization]. AB - The introduction and widespread application of Assisted Reproductive Techniques (ART) have raised major concern about the offspring 's health. The incidence of congenital and chromosomal anomalies after standard In Vitro Fertilization and Embryo Transfer seems to be similar to that expected in the general population. The prevalence of congenital malformations does not seem to be higher in children conceived by ICSI. On the other hand, it seems that there is a slight risk for transmission of chromosomal aberration of paternal origin and a certain risk of de novo sex-chromosomal and structural aberrations after ICSI. We report the results of the follow-up of 938 children conceived in our public ART by standard IVF(649) and by ICSI(289) from 21-2-1987 to 30-6-1999. The incidence of the congenital malformations results of the 1.8% (17/938); the incidence of chromosomal anomalies results 0.5% (5/938). The incidence of congenital malformations and chromosomal anomalies results 1.5% (10/649) and 0.6% (4/649), respectively, for standard IVF and 2.4% (7/289) and 0.3% (1/289) for ICSI. Our data seems to be reassuring but the incidence of chromosomal anomalies in ICSI children needs further investigation. PMID- 11424792 TI - [Various features of neonatal outcome in assisted fertilization pregnancies]. AB - Assisted pregnancies have been often considered as high risk pregnancies because of several factors as increased maternal age, primary sterility, pharmacological treatments, embryos and gametes manipulation in the first weeks of life. Many studies showed high rates of perinatal morbidity (especially low birth weight and prematurity), and an increase of maternal morbidity and mortality. The present study was designed to analyse correlation between morbidity and twinning rate, and to show the distribution of birth weight in a population of neonates after assisted reproductive techniques (ART). We studied 352 newborns (168 singletons, 126 twins, 42 triplets, 16 quadruplets) from assisted pregnancies; morbidity was 7.1% in singletons, 11.9% in twins, 21.4% in triplets, raising to 56.3% in quadruplets. Our experience suggests that multiple gestations after ART seem to have an increased incidence of neonatal complications. PMID- 11424793 TI - [Congenital cardiopathy in a data-based population]. AB - Altogether 429.139 consecutive births were surveyed during the eighteen years study period by the Emilia-Romagna Registry. Among these, 2147 newborns with congenital heart defects (CHD) (prevalence 5 per 1000) were detected within the first week of life. There were 1607 isolated CHDs and 540 cases had other associated defects. During the study period an increase in prevalence/rate of CHDs was observed (from 3.1 per 1000 in 1980 to over 7 per 1000 in 1998), particularly isolated CHDs increased from 2.2 to 6 per 1000, while the prevalence rate of CHDs cases with other associated anomalies was constant ranging from 1 to 2 per 1000. The increase of isolated CHDs was due to the increased number of "minor" lesions such as ventricular (VSD) and atrial septal defect. The apparent increase in birth prevalence of CHD mainly results from improved diagnosis due to widespread use of color-doppler ecocardiography. As in other studies, a significant shift in the sex-ratio has been documented: a male predominance in transposition of great arteries, left hypoplastic heart and aortic stenosis (male/female ratio 2.2, 2.3, 4.5 respectively) were found; while VSD had a slight female excess (male/female ratio 0.96). The study confirmed that the majority of the affected parents were mothers. The recurrence risk of a cardiac defect in first degree relatives was 2.3%, while the recurrence risk of isolated conotruncal defects was 3.9%. Out of 47 cases with isolated conotruncal defects 4 had microdeletion of chromosome 22q11.2 and concerning the first degree relatives in one case the father had the deletion without CHD. PMID- 11424794 TI - [Prevalence and distribution of congenital cardiopathy at birth and in pregnancy termination: impact of prenatal diagnosis in 4 years of experience]. AB - OBJECTIVE: To evaluate in two time intervals the potential impact of prenatal diagnosis on prevalence and spectrum of CHD at birth and in aborted fetuses. PATIENTS AND METHODS: At the University Hospital of Bari, south-east Italy, in the period between January 1st 1996 and December 31st 1999 a retrospective study was performed of all newborns and termination of pregnancy (TOP) beyond 18 weeks' gestation with postnatally diagnosed CHD. An antenatal fetal ultrasonic evaluation had been always performed between 18-22 weeks' gestation. The prevalence, distribution and detection rate of CHD at birth and in TOP were assessed and compared between two different periods of time (96-97 vs 98-99) to verify an eventual improvement in the prenatal diagnosis due to the learning curve and to new technologies. RESULTS: Prevalence of severe CHD in livebirths and aborted fetuses showed no significant changes between the two study periods (respectively 4.6@1000 vs 5.4@1000, at birth; 10% vs 11%, in TOP) and the same was observed as for as distribution of CHD. Maternal or fetal risk factors were found in 23% of cases of CHD at birth and in 74% of cases of CHD in aborted fetuses. The antenatal detection rate of CHD did not change between the two study periods in newborns (25% vs 27%, NS) while in aborted fetuses it was higher and showed a significant longitudinal improvement (53% vs 85%, p < 0.05), which was more evident if only selected cases were considered (56% vs 93%, p < 0.03). DISCUSSION: Our data show that gain in experience in fetal echocardiography has increased the prenatal diagnostic accuracy for congenital cardiac malformations only in selected cases, evaluated by more expert operators. In conclusion the impact of antenatal routine screening for congenital heart disease appears still relatively small. PMID- 11424795 TI - [Registry of genetic and malformative diseases in the province of Macerata]. AB - OBJECTIVES: The aim of this epidemiological research is to evaluate the prevalence of genetic diseases and malformative syndromes in paediatric population living in the Macerata county. MATERIAL AND METHODS: All the data were collected through a careful analysis of a specific questionnaire sent to all the family paediatricians. RESULTS: 23,379 children living in Macerata county, aged 0 to 9 years, were evaluated (93.8% of all this paediatric population). Among those were found N 400 cases of genetic diseases and malformative syndromes: Malformations Tot.N. 255 cases (63.3% of the reported cases); Malformative Syndromes Tot. N. 55 cases (27.8% of the reported cases); Endocrinology and Metabolic Diseases Tot. N. 41 cases (10.3% of the reported cases); Osteochondrodysplasia Tot. N. 22 cases (5.7% of the reported cases); Other Tot. N. 28 cases (7.0% of the reported cases); Male population was found more affected than female: M/F ratio = 1.4. The analysis of the data showed an increasing trend in detecting these pathological conditions, consistent with the increase in geographic altitude (3 areas considered): 0-100 meter = 0.88%; 100-600 m.a.s. = 1.34%; over 600 m.a.s. = 1.88%. CONCLUSION: The knowledge of the number of children affected by genetic and malformative diseases in the Macerata county is relevant in order to establish a Genetic Service with the aim to better support the medical assistance of these patients and counselling service for the families. PMID- 11424796 TI - [Clinical features in subjects with congenital anomalies of the corpus callosum]. AB - The corpus callosum is a connecting structure between the two cerebral hemispheres. Its development occurs between 8th and 20th week of gestation. Anomalies of the corpus callosum are divided into malformative or acquired anomalies and may be isolated or associated with other cerebral malformations. The diagnosis of an alteration of the corpus callosum is fundamentally based on neuroradiological examinations. In our study we evaluate the major clinical findings, the psychomotor development, the electroencephalographic and neuroradiological features in a group of 14 children with congenital callosal anomalies, trying to single out how the associated cerebral malformations interfered with the clinical manifestations and especially the prognosis. It was not possible to highlight a distinctive symptom indicative of the presence of a corpus callosum anomaly; nevertheless in 11/14 children there was a psychomotor delay of varying degree. 42% of our subjects presented epilepsy, with a typical West syndrome in 5/6. Children affected by isolated callosum anomalies did not present any significant clinical manifestations. Finally, we conclude that the determinating factors in the severity of the clinical-electroencephalographic picture as a whole, and especially in the prognosis of our patients are the cerebral malformations associated to the callosum anomalies. PMID- 11424797 TI - [Neurologic outcome in infants surgically treated for congenital cardiopathy: preliminary data]. AB - INTRODUCTION: Both surgical techniques for correction of congenital heart diseases (CHD) and intraoperatory neurologic protection improved during the last 20 years. Nevertheless cardiac surgery is still a risk for neurologic morbidity. METHODS AND PATIENTS: Analysis of the postoperative neurologic status of infants younger than 6 months who underwent cardiac surgery from January 1998 to December 1999. We reviewed the EEG tracings, cranial ultrasound reports (CUS) and CT scans of 48 patients. Diagnoses were: ventricular septal defect = 15, Fallot (TOF) = 9, patent ductus arteriosus (PDA) = 5, coarctation of aorta = 4, atrio-ventricular septal defect = 4, transposition of great arteries (TGA) = 3, hypoplastic left heart syndrome = 2, pulmonary atresia = 2, total anomalous pulmonary veins drainage = 2, double outlet right ventricle = 1, cor triatriatum = 1. Mean age (range) at intervention was 54 days (2-150), 44 infants (91.7%) survived at follow-up: 23 EEG, 22 CUS and 2 CT were performed in the recent postoperative. Among survivors 5/44 had neurologic complications. EEG was altered in 4: two of them (1 TOF, 1 TGA) had pathologic CUS and CT as well (ischemic pattern in the former, atrophy in the latter). Finally a preterm newborn with PDA had mild abnormalities at CUS. After a mean follow-up of 16 +/- 6 months 3/5 patients had mild-to-moderate psychomotor delay and 2 recovered. CONCLUSIONS: According to our preliminary data the prevalence of neurologic complications in infants who undergo cardiac surgery seems to be low. The pathological findings of the recent postoperative seem to recover up to normalization in some cases at mid-term follow-up. As expected, permanent complications effect more often complex CHD. Further follow-up studies to school age will be mandatory to check the very final results of cardiac surgery performed during early infancy. PMID- 11424798 TI - [Congenital malformations and asymmetric crying facies]. AB - Asymmetric crying face (ACF) means a congenital anomaly caused by either agenesis or hypoplasia of the depressor anguli oris muscle. This defect is on only one corner of the mouth since the birth, affects lower lip, and is particularly evident when the newborn is crying. Lesions at different levels of seventh nerve can cause similar and confounding weakness of the facial expressions. But, in case of ACF, forehead wrinkling, eye closure, nasolabial fold depth, and tearing are normal and symmetric functions. ACF can be either as single plain aesthetic defect or as early only index of several congenital malformations, especially of heart and genitourinary tract. Our study recognized 80 ACF cases in 11,643 newborn's population during a 34 consecutive month period: 34 on right side and 46 on left side of the mouth. ACF population presented a higher malformative risk than general population (on average 4.73% versus 3.3% of our base-line). Congenital anomalies were found more frequently associated with left ACF (3 versus 1, for a general rate of 6.52% versus 2.94%), on the same body side when anomaly was affecting pair organs. Those 3 congenital malformations were on genitourinary tract, 2 of which detected by ultrasonography. We did not find any congenital cardiac defects. According to opinion of numerous other researchers, we think ACF is not to be considered a simple aesthetic anomaly: therefore, in front of all ACF cases, a thorough search for associated anomalies should be performed. PMID- 11424799 TI - [Cervico-inguinal microbiology, vaginal pH, infections, and premature labor]. AB - Preterm labor is the leading cause of perinatal morbidity and mortality. There is increasing evidence that inflammation of the upper genital tract, deriviated by lower reproductive tract infections (cervicitis or vaginitis), may play a major role in the pathogenesis of preterm labor, premature rupture of membranes and preterm birth. These complications are potentially preventable with prompt diagnosis and treatment in pregnancy. Alterations of the vaginal ecosystem with change in vaginal pH, are presupposition for infections and their consequences. OBJECTIVE: To investigate the relationship between cervico-vaginal infections and related events such as expression of bacterial enzymes and change in vaginal pH with preterm labor. METHODS AND RESULTS: In our study we identified cervico vaginal infections during gestation week 8-14 and 30-34, using three different diagnostic tools: 1) microbiological tests; 2) assessment of vaginal pH; 3) detection of bacterial enzymes. Then, we correlated these events to perinatal outcome. Among 200 pregnant women a pH greater than 4.6 was detected in 38% of all cases. Among microbiologically positive patients, vaginal fluid pH was positive, in particular, in 75% cases of Bacterial vaginosis. CONCLUSION: pH assessment along with symptoms and signs could be used as initial screening test to alert physician to vaginitis. When vaginal fluid pH is used to screen for Bacterial vaginosis, it has a greater sensitivity than when used to screen for all causes of vaginitis. PMID- 11424800 TI - [Microbiological monitoring in neonatal intensive care]. AB - There is no "ideal" method of infection diseases control, but there are some different methods according to the risk degree of patient, the problems, the information accuracy and also the available resources. Some important points need to be stressed about infectious diseases control in Neonatal Intensive Care Unit: -continuous microbiological surveillance in Neonatal Intensive Care Unit is necessary, because newborns admitted are at very high risk of infection disease;- continuous microbiological data review is needed: therefore criticism and integration with other clinical and laboratory data are necessary to exclude simple colonisation;--critical analysis of data allows more rational use of antimicrobical agents to avoid the selection of multiresistant bacteric streams;- a rational analysis of laboratory data needs necessarily a strict collaboration among neonatologists, microbiologists and the Committee for infectious diseases control;--in epidemics, techniques of bacterial streams typization are needed, and methods of molecular biology (DNA and proteins analysis) are better;--this implies a collection of microbiological data, by database integrating microbiological, clinical and anamnestic data and allowing retrospective studies too. PMID- 11424801 TI - [The false myth of toxoplasmosis in salami]. AB - On the basis of available literature concerning Toxoplama gondii infection in swine (swine toxoplasmosis) and the resistance of parasite to heating, freezing, gamma-radiation, salting, seasoning and production processes of "cooked" and "fermented" salami, the possible risk of transmission of the disease from pig (where toxoplasmosis is widely diffuse) to man is discussed. No risk of transmission of the disease, even to children and pregnant women, can derive from the consumption of cooked meat, cooked or salted-fermented salami and ham. The only risk could be related to eating of even small amount of fresh sausages or chitterlings (especially heart, brain and other viscera) and consumption of not drinking water. PMID- 11424802 TI - [Risk assessment of vertical transmission of Toxoplasma infections]. AB - BACKGROUND: It is important to estimate the risk of congenital toxoplasmosis infection and its clinical sequelae. This information will assist the clinical counselling of pregnant women with acute toxoplasmosis and may guide individual decisions on investigative and therapeutic options. METHODS AND PATIENTS: Since January 1998 to July 2000 we collected data on 63 pregnant women referred to our Prenatal Diagnosis Centre for suspicion of acute maternal toxoplasmosis. In case of positive screening tests for IgG and IgM or documented seroconversion, we sent samples of maternal serum to a reference laboratory to detect specific IgM, IgG, IgA antibodies and IgG avidity. We estimated the risk of mother-to-child transmission as high or low in relation to gestational age at the time of maternal seroconversion. Antiparasitic treatment with spiramycin since before confirmation of infection and evaluation of fetal biometry were done. In high risk women we advised amniocentesis for polymerase chain reaction (PCR). If infection was diagnosed in the fetus, the regimen of treatment was changed. Live born children of all these mothers were followed up serologically for at least 1 year. RESULTS: In 13/63 cases (20%) we excluded maternal infection. Thirty-six out of 63 cases (57%) with high transmission risk underwent an amniocentesis, except one case requesting termination of pregnancy. Four out of the remaining 14 cases estimated at low risk had an amniocentesis for maternal anxiety. Among high risk women, PCR on amniotic fluid was positive in 4 cases (10%): 2 of these requested pregnancy termination, while the other 2 decided to proceed with the pregnancy and the therapy was shifted to 3 weeks courses of pyrimethamine and sulphadiazine with folinic acid alternating with spiramycin. Five infants were lost at follow-up, 20 babies resulted not infected and the remaining 21 are still followed-up. CONCLUSION: It is possible to select by specific serological tests, high risk cases for mother-to-child transmission of infection and to decide a specific approach. PMID- 11424803 TI - [Prophylaxis of group B beta-hemolytic streptococcal infections]. AB - Group B streptococcus is a possible cause of chorioamnionitis, endometritis and urinary tract infections in pregnant woman. Maternal risk factors and the vertical transmission of GBS and neonatal GBS infection occur through the following fever during labor, the rupturing of membranes more than 18 hours before delivery, prematurity and chorioamnionitis. GBS can induce early-onset neonatal disease (sepsis, meningitis or pneumonia) during the first week of life and late-onset neonatal infection (leptomeningitis) within the first 12 weeks of life. Numerous strategies for preventing neonatal group B streptococcal infection were investigated: 1) the treatment of GBS-colonized women during the third trimester of pregnancy did not prove to be effective because it does not reduce maternal colonizzation rates at delivery; 2) the neonatal universal post-partum prophylaxis with penicillin G was ineffective and increased neonatal mortality due to penicillin-resistant bacterial infection; 3) the intrapartum maternal chemoprophylaxis with penicillin G or ampicillin in GBS-colonized women, in women with risk factors, or in women with both GBS colonization and risk factors. The latter strategy proved to be the most effective because it reduces the risk of early-onset GBS infection by 75% and 95% when associated with post-neonatal prophylaxis. To date, there are no guidelines on the management of the asymptomatic neonate whose mothers have been treated with chemopropylaxis intra partum. PMID- 11424804 TI - [Therapy or prevention of fetal infection by cytomegalovirus with immunoglobulin infusion in pregnant women with primary infection]. AB - OBJECTIVE: Administration of cytomegalovirus (CMV)-specific immunoglobulins to pregnant women with primary CMV infection in order to inhibit viral activity. MATERIALS AND METHODS: We considered 2 groups of patients including 24 pregnant women. GROUP A: 12 women with primary maternal-fetal CMV infection, shown by CMV culture and CMV DNA detection in 9 (75%) and by only CMV DNA detection in 3 (25%) of the amniotic fluid (AF) samples. These pregnant women were treated with infusions of CMV-specific immunoglobulins (200 U/Kg of maternal weight and 400 U/Kg of fetal weight to prevent CMV pneumonia and gastroenteritis). As control group we considered 15 pregnant women, 5 of whom had CMV-positive AF samples demonstrated by polymerase chain reaction (PCR) and 10 also by CMV culture. GROUP B: 12 women with primary CMV infection, who were treated with monthly infusions of specific immunoglobulins to prevent the transmission of CMV to the fetus. In the control group we followed up 53 patients who were not treated, including 15 subjects with CMV-positive AF samples, 20 with CMV-negative AF samples and 18 women who did not accept amniocentesis. RESULTS: GROUP A: All 9 neonates born to mothers with culture and DNA positive AF samples were CMV infected. On the contrary, the babies born to 3 women with only PCR-positive amniotic fluid were CMV-negative by culture and DNA detection. Of 9 neonates infected, 8 were asymptomatic and 3 became culture-negative before they were one year old. The only symptomatic baby (IUGR and ventriculomegaly diagnosed by ultrasound during the pregnancy) was treated with ganciclovir and foscarnet. Of 15 non-treated patients, 11 had ultrasound signs of placental and/or fetal CMV involvement. Seven of these (46%) aborted and 4 (27%) delivered neonates with severe symptomatic infection. The other 4 patients delivered oligo-symptomatic CMV infected neonates positive by culture or PCR only. Overall, the prevalence of symptomatically infected neonates or fetuses was significantly higher (p = 0.02) among non-treated than treated women. Moreover, the babies of treated women showed less prolonged (p = 0.01) viruria than those of non-treated patients. GROUP B: all 11 pregnant women, treated with immunoglobulins delivered CMV negative neonates. One patient, who had interrupted infusions at the 24th gestation week, delivered an asymptomatic CMV infected baby. In the control group, 4 women with CMV-negative AF samples delivered neonates with asymptomatic infection. Of 18 patients who did not undergo amniocentesis, 9 (50%) aborted; of remaining 9 women, 3 (33%) delivered CMV infected neonates. CONCLUSIONS: Our data suggest that CMV immunoglobulins may be effective for treatment or prevention of fetal CMV infection. PMID- 11424805 TI - [Pregnancy in women infected with the hepatitis C virus]. AB - The prevalence of HCV-RNA positivity in pregnant women goes from 1.2% to 4.5% in different countries. The aim of our study is to show pregnancy outcome, vertical/perinatal transmission rate, the viral load and the tramnsaminases trend during pregnancy and after delivery. The study involved 11,681 pregnant women screened in the Obstetric Department for High Risk Pregnancy at the University of Padua between 1992 and 1999. We evaluated the markers of HCV, HBV, HIV, the viral load and genotype and AST/ALT in the mothers and positivity and viral load of HCV RNA in the newborns at birth and at 3rd, 6th, 9th, 12th month. Of the 11,681 pregnant women, 135 (1.15%) tested positive for the presence of anti-HCV antibodies and of the 135 anti-HCV antibody-positive mothers, 80 were found to be positive for HCV-RNA. Of 80 pregnancies that were HCV-RNA positive, 4 termined in abortion, 1 in stillborn, 1 in neonatal death, 18 in preterm delivery and 56 were carried to term. We came to the conclusion that HCV infection does not increase the risk of obstetric complications and does not influence the fetal-neonatal status at delivery; the pregnancy evolution may be complicated by the onset of cholestatsis in the 2nd and 3rd trimester; vertical transmission of the infection develops in few cases (4.8%), more likely at delivery. PMID- 11424806 TI - [Vertical transmission of the hepatitis C virus]. AB - The incidence of hepatitis C in childhood is approximately 0.4%. The mode of transmission can be parenteral, sexual, occupational and also vertical. The latter has an incidence that varies widely and it increases in the case of human immuno-deficiency virus (HIV) coinfection and high titers of HCV in the mother. The vertical transmission is not influenced by breast feeding, however, data are discrepant with regard to child delivery (cesarean section vs vaginal delivery). Ninety-seven babies born from mothers with hepatitis C from 1996 to 1999, were evaluated prospectively in the Day Hospital of the Pediatric Department of Parma. The protocol of observation established a blood sampling for titers of antibodies anti-HCV and HCV-RNA at the 3rd trimester of pregnancy and subsequent clinical and biochemical controls at 3-6-9-12-15 and 18 months. Thirty (31.2%) out of the 96 mothers evaluated were positive for antibodies anti-HCV and 66 (68.8%) were positive for antibodies anti-HCV and HCV-RNA. Five (5.15%) out of the 97 babies evaluated were infected by HCV. Of these 4 were delivered vaginally and 1 by cesarean section. Of the 3 babies born to mothers with HIV coinfection, none was infected by HIV, but 1 was infected by HCV. Vertical transmission is increased by HCV viral load or HIV coinfection in the mother. The vaginal delivery and breastfeeding do not represent an additional risk factor. PMID- 11424807 TI - [Risk of materno-fetal transmission of the HIV infection with antiretroviral therapy and cesarean section: experience of the Parma group]. AB - Vertical transmission of HIV is by far the most important way of infection in pediatric patients. Transmission rate of infection varies between 15-40% in the absence of antiretroviral prophylaxis. Only 2% of infected pregnant women who underwent caesarean section and zidovudine treatment transmitted the infection to their newborns. From January 1995 to September 2000 twenty seropositive pregnant women and their twenty newborns were followed at the Azienda Ospedaliera of Parma. Nine women (45%) were treated with only zidovudine according to the ACTG 076 protocol; eight women (40%) continued the treatment they were assuming before pregnancy with the eventual addition of zidovudine. 3 women (15%) were not treated because HIV infection was only detected after delivery. 15 women underwent caesarean section, in 13 cases in association to antiretroviral prophylaxis: in the remaining 2 cases no intrapartum treatment was started due to the urgency of delivery. The rate of vertical transmission among the 20 women was 5% (1/20), significantly less then that observed (20.5%) among 31 pregnant HIV women followed in Parma from January 1987 to December 1994 and not treated with antiretroviral prophylaxis and/or cesarean section (Magnani G. Personal data). The only infected baby was born by vaginal delivery. No transmission was observed in the group of pregnant women who underwent the combination of antiretroviral prophylaxis and cesarean section. PMID- 11424808 TI - [Lavage of the birth canal with chlorhexidine: a new valid method for the prevention of perinatal infections]. AB - Perinatal morbidity and mortality are due to various infective agents, mainly represented by beta-hemolytic group B Streptococcus. The perinatal disease related to this infection is distinguished in Early-onset, characterised by pneumonia and sepsis, and Late-onset which leads to sepsis, meningitis and pneumonia. Various strategies were therefore proposed to prevent transmission including immunisation and chemoprophylaxis. All these methods however present adverse effects and are most of all expensive to carry out. Taha et al. reported an interesting experience regarding the reduction of perinatal infections following the cleansing of the birth canal with a solution of Chlorhexidine 0.25% during labour (1996-1997). It seemed interesting for us to assess the applicability and efficacy of a new strategy of prophylaxis of perinatal infections in a Developing Country based on the association of two of the simple strategies proposed i.e.: cleansing the birth canal with chlorhexidine and chemoprophylaxis in cases with risk factors without culture screening. We studied two groups of patients: one in which cleansing of the birth canal was used and the second (control group) in which the old method already applied in the hospital (i.e. cleansing of the external genitals with Cetrimide 1%+ Chlorhexidine 0.1%) was carried out associated with antibiotic therapy when risk factors arose. We observed a total absence of neonatal mortality due to sepsis resulting from the association of the methods suggested even though the presence of sepsis evaluated through signs and symptoms like fever, poor feeding, apnoea or dyspnoea in newborns was similar in both groups. PMID- 11424809 TI - [Use of palivizumab in the prevention of syncytial virus respiratory infection]. AB - BACKGROUND: Monthly intramuscular Palivizumab (Synagis, MedImmune, Inc.) is effective in reducing the incidence of RSV-attributable hospitalization by 55% if compared with placebo and seems to be well tolerated. METHODS: Our clinical experience in the use of palivizumab in RSV-prophylaxis is presented. During autumn-winter 1999-2000, a total of 8 newborns (gestational age between 28-34 weeks) in our neonatal intensive care unit (NICU) met the AAP recommendations for RSV-immunoprophylaxis. Palivizumab was monthly administered at a dosage of 15 mg/kg i.m. at discharge from the NICU and during the RSV season. Infants were followed up to 40 days from the last injection for adverse reactions and clinical data. RESULTS: Among eligible newborns, two (28 and 29 weeks respectively) were affected by CLD and required medical therapy at discharge, 4 were born at 29-32 weeks and 2 (both of them born at 34 weeks) had additional risk factors of infection (crowded environment, passive smoking). During RSV season, none of the infants experienced RSV-hospitalization nor lower respiratory tract infection. Mild and transient adverse effects occurred in 2 cases out of 8 (induration in the site of injection and irritability). CONCLUSIONS: In our series of patients, palivizumab resulted safe and effective. PMID- 11424810 TI - [Role of Parma in the nutrition culture of the country and of Europe (the protection of the European Union)]. PMID- 11424811 TI - [Extra-nutritional and nutraceutical effects of traditional foods of Parma]. AB - Territorial Products and food-culture relationship are described. On the basis of food extra-nutritional constituents and on the characteristics of functional food (nutraceutical food), traditional Parma products, i.e. Parma Ham and Parmigiano Reggiano Cheese, are examined in their nutritional, extra-nutritional and functional consequences. PMID- 11424812 TI - [Food additives: effects on pregnancy and lactation]. AB - Exposure to several substances in a critical period of life, as in perinatal age, could cause long-term effects. Among these agents we can consider food additives, defined as "substances used to conserve, colour or flavour ailments without nutritional effects". Food additives utilisation by women during pregnancy and breastfeeding has not to be considered completely safe; actually, current evidence suggests that these substances induce alterations of the differentiation of several cell-types, resulting in development of disease during the adult age. During pregnancy, for instance, E100 color additive, is suspected to be embryotossic, amaranth causes a reduced olfactory orientation in F1 generation mice and beta-cyclodextrin produces a transient neonatal growth retardation in rodents. The food additive methylmercury is excreted in human milk and causes neurotoxicity in breastfed babies. Steroids pass in human milk, too: they can inhibit lactation and cause newborn breast ipertrophy. Regulations to avoid these early exposures may contribute to an important improvement of health conditions of humankind. PMID- 11424813 TI - [Pre-eclampsia: role of excessive caloric intake]. AB - OBJECTIVE: To compare the average daily diet of women having physiologic pregnancy versus patients presenting preeclampsia (PE) in order to evaluate if increased caloric intake is associated with preeclampsia. METHODOLOGY: The macronutrients and kilocalories in measurements obtained from diaries filled in over a seven day period by 147 pregnant women (30 preeclamptics, 117 with physiologic pregnancy were compared (the proportion of about 1/4 between the two groups was chosen to increase the statistical efficacy, because this pathology is rather unfrequent). RESULTS: The analysis of the diaries shows that the preeclamptic women consume, every day a greater quantity (p < 0.05) of kilocalories, proteins and lipids than the physiologic pregnancy. No statistically significant difference was found for fibres and carbohydrates (the last ones recorded without distinguish mono and disaccharides from polysaccharides). Physiologic pregnancy mean Body Mass Index is 21.6 versus 26.4 of PE women (p < 0.05). CONCLUSION: Intake of energy, protein and lipid were positively associated with risk of PE. Overweight patients are highly at risk of contracting PE. These data, according to others trials confirm the advisability: 1) to analyse the diet of pregnant women; 2) to reduce nutritional intake, if in excess, first of all, in women at risk of PE; 3) to suggest loosing weight to overweight patients before initiating a pregnancy. PMID- 11424814 TI - [Cerebral oxygenation and near-infrared rays spectrophotometry]. AB - Near infrared spectroscopy (NIRS) is a new technique which allows non invasive bedside monitoring of cerebral oxygenation and hemodynamics by measuring relative changes in cerebral oxy- and deoxyhaemoglobin and cytochrome aa3. We have applied this technique to evaluate the possible effects on cerebral oxygenation and hemodynamics of clinical procedures usually performed on preterm infants:- endotracheal suctioning, and we have demonstrated that the magnitude and the duration of the negative effects of open system are significantly reduced using closed endotracheal suctioning system;--withdrawal and infusion through umbilical vein and artery cause significant changes in cerebral hemodynamics: these effects are significantly reduced after administration of ibuprofen;--treatment of patent ductus arteriosus with ibuprofen does not significantly reduce cerebral perfusion and oxygen availability compared to indomethacin and ibuprofen administration also does not affect cerebral vasoreactivity to arterial carbon dioxide tension;- administration of different types and doses of natural surfactant causes different changes in cerebral hemodynamics and these effects seem to be dose related. Therefore NIRS is an useful device to investigate cerebral oxygenation state of preterm infants and new possibilities could derive from the introduction of a new NIRS method which allows to measure the tissue oxygenation index. PMID- 11424815 TI - [Evaluation of cerebral oxygenation in newborns with prematurity apnea: new frequency domain NIR oximeter]. AB - Near infrared spectroscopy (NIRS) is a non invasive, portable, safe technique for monitoring cerebral oxygenation and haemodynamics. A new frequency-domain tissue oximeter based on a multi-distance measurement protocol is presented. The effects of apneic episodes on cerebral and peripheral arterial oxygen saturation (SatO2) in preterm newborns, as monitored by NIRS and by pulse oximetry, are reported. The study population consist of 5 preterms (26 to 30 weeks of gestational age), in the second week of postnatal age, affected by apnea of prematurity. NIRS and pulse oximetric measurements were made contemporarily for a 40-minutes period for each infant. All monitorized apneic events were associated with bradicardia, and resolved spontaneously or after tactile stimulation. As results: a) there was always cerebral deoxygenation in association with apneic events, b) the mean SatO2 as measured by NIRS was slightly lower than the pulse oximeter readings, c) cerebral SatO2 decreased faster and the absolute value of the cerebral SaO2 decrease was greater than that measured peripherally (mean value of 27 versus 13%), d) increases of cerebral deoxyhemoglobin and total hemoglobin and a decrease of oxyhemoglobin were also observed. These preliminary results show that peripheral oxygen saturation measurements as measured by pulse oximetry could not always reflect brain oxygenation. PMID- 11424816 TI - [Retinopathy of prematurity: incidence and risk factors]. AB - The study analyses the incidence of the retinopathy of prematurity (ROP) and its correlation with the most important risk factors in neonates with gestational age (GE) < or = 32 weeks and/or with birth weight (BW) < or = 1500 g. The cohort of our study is composed by 305 preterms with Mean GE of 29.8 weeks and Mean BW of 1312 g, studied at the Neonatal Intensive Care Unit of the University of Parma during January 1993-December 1999. The incidence of ROP resulted 19.7% in our group and was inversely proportional to the GE and BW. No preterms with GE < or = 32 weeks presented retinopathy, while there has been an incidence of the 65% in the subjects with GE lower than 26 weeks. The most severe grade of ROP was mainly seen in the newborns with the lowest GE (31%), whilst only the 1.5% of the preterms with GE of 30-31 weeks developed ROP of stage 3 or 3+. Similar results were seen when the data were compared with the birth weight. However a direct relation was seen with the duration of the O2 therapy and with the typical pathologies of the prematurity, namely respiratory distress syndrome (RDS) and intraventricular haemorrhages (IVH). Among the other risk factors evaluated, the blood transfusion was the most relevant. PMID- 11424817 TI - [2,3 diphosphoglycerate in preterm newborns]. AB - BACKGROUND: It has been largely shown that during the first month of life, in the preterm neonate Hb levels and Hct percentages rapidly decrease, high HbF concentration persists and a high oxygen affinity occurs. Data are needed to establish the level at which 2,3 dyphosphoglycerate (2,3 DPG) interacts with the regulation of oxygen affinity. SUBJECTS AND METHODS: 24 samples, from eight uncomplicated preterm newborns (34.1 +/- 1.83 GW, 1869 +/- +/- 291 BW) obtained at the same time as those required for the clinical management of the infants, were collected on the 2nd, 7th and 14th day of life. Blood gases, total hemoglobin and hematocrit were obtained from 0.3 ml arterialised capillary blood. Assays of 2,3 DPG were made separately on 0.4 ml venous blood. RESULTS: As expected tHb concentration and Hct percentages significantly decreased from day 2 to day 14 in all eight cases. On the contrary 2,3 DPG and p50 values remained stable. Subsequently throughout the study period all neonates had an increased 2,3 DPG/Hb ratio that was significantly related with p50 at standard conditions (p < 0.05). CONCLUSIONS: Stable 2,3 DPG concentrations during all study period have been detected. The subsequent significant increased 2.3 DPG/Hb, ratio related to increased p50 values, could have a key role in a physiological mechanism aimed to ensure adequate oxygen delivery to the tissues and to counteract the higher oxygen affinity of fetal hemoglobin. A wider sample is needed to validate this hypothesis. PMID- 11424818 TI - [Use of recombinant human erythropoietin in newborns with very low weight: efficacy monitoring]. AB - Chronic anemia is very frequent in very low birth weight (VLBW) infants. Lowered red cells life span, hemolysis, low production of erythropoietin, phlebotomies, excessive body growth are its most important causes. A reduction of the number of transfusions to babies with chronic anemia was obtained through r-HuEpo. A serie accounting for 89 newborns < 1500 g (18 < 1000 g) with a mean weight of 1069 g (+/- 238) in whom early treatment with r-HuEpo was performed (from 9.55 +/- 3.04 day), 300 UI three times a week s.c., is presented. Therapy with r-HuEpo was carried out for 6 weeks, or until the baby weighed 1800 g. During the treatment, each baby received iron, folic acid, multivitaminic supplements. Patients were monitored with red blood cells count, comprehensive of reticolocytes, ipochromic cells (Ipo-cells), content of hemoglobin of reticolocytes (CHr), each week. Iron, ferritine and transferrine were obtained only twice a month, as they required further blood sampling. 10.1% neonates received transfusions: the percentage of transfused VLBW infants was much higher (55.5%) before than after the introduction of r-HuEpo (p = 0.0002). 33.3% extremely low birth weight (ELBW) infants required transfusions (vs 95.5% in pre r-HuEpo period) (p < 0.0001). Our results confirm the importance of Ipo-cells and CHr to monitor early alterations of iron cellular employment. PMID- 11424819 TI - [QTc interval in newborns with gastroesophageal reflux treated with cisapride and ranitidine]. AB - Gastroesophageal reflux (GER) is very common in infants, especially in prematures and may be cause of gastrointestinal and cardiorespiratory symptoms. Cisapride, a prokinetic agent, is used in order to avoid the transient esophageal sphincter relaxation, but it is sometimes associated to transient prolongation of QT interval on EKG, especially with high dosage. The authors report the effects of cisapride therapy (0.8 mg/Kg/day) on QTc interval (QTc = QT interval corrected on heart frequency) in a pediatric population (50 infants) with GER. Results demonstrate the relatively safety of cisapride therapy at low dose also in the pediatric period. PMID- 11424820 TI - [CRIB score: mortality, morbidity, and long-term neurologic development]. AB - The purpose of this study is to assess the possibility of predicting mortality, major pathology and long-term neurodevelopmental impairment in very low birth weight VLBW infant using Clinical Risk Index for Babies (CRIB). We studied a cohort of 251 VLBW infants, whose CRIB could be calculated, born from 1995 to 1998 in our Unit. We analyzed the mortality before discharge, the incidence of chronic lung disease (CLD) and of retinopathy of prematurity (ROP), the length of stay before discharge and the neurodevelopmental impairment at one and two years of corrected age using the Griffiths developmental scales (impairment was defined by a general quotient of 85 or below). The CRIB score was divided into three risk groups: 0-5, 6-10 and > 10. Mortality rate raises with the CRIB's increase (respectively 5.6% in the first group, 32.4% in the second and 93.8% in the third group); besides the incidence of severe ROP and of CLD, calculated in infants survived > 28 days, is higher (18.4% and 40.7% respectively) in the second group than in the first (1.9% and 7.4% respectively). In the end, the incidence of neurodevelopment impairment at one and two years of corrected age is respectively 6.8% and 6.0% for children with CRIB 0-5 and 29.4% and 21.4% for children with CRIB 6-10. CRIB score is strongly associated with mortality and there is an increasing risk for severe ROP, CLD and neurodevelopment impairment from class 0 5 to class 6-10; no statement can be made for these diseases in class > 10 because there is only one survivor in this class. PMID- 11424821 TI - [Percutaneous catheterization of the newborn: control of pain and easy insertion with brachial plexus block]. AB - The increasingly prolonged survival of extremely premature infants who required long-term venous access means that this is often a major problem in modern neonatal unit. The insertion of central venous catheters has become an established practice and the development of silastic catheters inserted by newer percutaneous techniques through a peripheral vein is, now, the choice technique in awake non sedated critically-ill infants. Such technique has an high percentage of failure. We have undertaken a retrospective study to determine whether the brachial plexus block performed via the axillary approach could improve the success rate for the insertion of central venous catheter from a peripheral vein of the upper limb in infants minimizing physical and emotional stress to the neonates. Data from 157 low and very low birthweight infants, submitted or not submitted to the axillary block, were examined. The failure rate for the insertion of the central venous catheter was 27% without using the brachial plexus block vs. 9% in the patients that underwent the block. Use of the brachial plexus block via the axillary route, although retrospectively evaluated, improves the success rate and the pain control for the insertion of small diameter central venous silicon catheter from a peripheral vein of the upper limb in low and very low birthweight infants. PMID- 11424822 TI - [Positioning of umbilical vein catheter with ECG-guided technique: randomized study]. AB - GOAL: To evaluate the effectiveness of electrocardiography-guided technique to aid in the correct positioning of umbilical vein catheters. DESIGN: A prospective, randomized controlled study. METHODS: Term and preterm newborns who required an umbilical venous catheter were managed by an ECG-guided technique (group A) or by a conventional method (group B). Correct positioning was defined by a chest-X-ray when the catheter tip was located above the diaphragm and outside the right atrium. For the ECG-guided technique we utilized a conductive device Vygocard (Medival, Padova) inserted in a 3-way stopcock connected with the catheter. The catheter was inserted under ECG observation until the appearance of a tall P-wave in lead III, which indicated the tip was within the right atrium. The catheter was then withdrawn until the P wave size returned to normal. RESULTS: We enrolled 44 patients (16 F, 28 M). Median gestational age (GA) and birth weight (BW) were 34 weeks (range 26-41) and 2130 g. (590-3870), respectively. Sex distribution, GA, BW and Apgar scores were not different between patients in group A (n = 22) and group B (n = 22). Catheters could not be advanced till the estimated insertion depth in 11 patients (A = 5, B = 6). In the remaining 33 patients, correct tip placement was more frequent in group A (88%) compared with group B (50%) (p = 0.021 by Fisher's exact test). No side effects specific to the ECG-guided method were noted. CONCLUSIONS: The ECG-guided technique seems to be a safe and effective method for the proper placement of umbilical vein catheters in newborns. PMID- 11424823 TI - [Digestive endoscopy in neonates. Personal experience]. AB - Gastrointestinal (GI) endoscopy is actually considered an essential diagnostic and therapeutic technique both in pediatrician and in newborn infants. At the moment upper GI endoscopy in neonatal age allows to explore the esophagus, stomach and duodenum; instead lower GI endoscopy easily reaches sigmoid-colon junction. The exploration of other distal parts of bowel is more difficult and with more risks. Therefore it has to be make only if it is necessary. In the newborn this exam should be so fast not to require premedication. In our study, we considered every neonatal patient who underwent GI endoscopy at the "Digestive Endoscopy and Gastroenterology Service of Pediatric Clinic-University of Parma" (from September 1983 to June 2000). We considered two groups of patients: first group: early neonatal age (0-14 days) second group: late neonatal age (15-28 days). We made 127 endoscopies: 112 of these were upper GI. The most frequent symptoms that babies showed and indicated upper GI endoscopy were reflux and/or vomiting (86%). Upper operative GI endoscopies included also 7 esophageal dilatations. The lower GI endoscopies were 15 and in all the cases the indication was intestinal bleeding. We conclude that the GI endoscopy in the neonates shows a diagnostic sensibility and specificity really high and for sure better than the radiology for mucosal lesions. Also GI endoscopy can be not only a very precious diagnostic instrument but also therapeutic in neonatal disease. Considering the age of the little patients, it is very important that only endoscopists with large experience use this technique. PMID- 11424824 TI - [Proposal for computer-assisted preparation of parenteral nutrition solutions in neonatal intensive care]. AB - A computer software package was developed to perform the calculation involved in compounding total and partial parenteral nutrition for low-birth-weight and sick newborns. The program requires a minimum of user input and is menu-driven. The flexibility of the algorithm has been increased to a considerable degree. The program calculates the overall balance of fluids, nutrients, calories, electrolytes and minerals. It allows for the possibility of total parenteral nutrition and simultaneous oral feeding. Nutrient amounts per Kg per day and combination of enteral food and parenteral infusions can be completely changed by the operator. In our experience computer-assisted system was more efficient than a manual system and the possibility of computational error was reduced. PMID- 11424825 TI - [Telemedicine in neonatal emergencies]. AB - Telemedicine is the transmission of medical notices and images among remote sites, that uses adequate audio-video systems. OBJECTIVE: To increase the quality and the amount of medical informations, avoiding unnecessary carriages. Among main fields of interest in Telemedicine, monitoring of cardiovascular parameters and medical emergency represent situations that need to be promptly and appropriately approached. In such a similar conditions, a preliminary transmission to III level Health Institutions of informations as Electrocardiograms or even Echocardiograms may play an essential role in the diagnosis, prognosis and treatment of emergent cardiovascular disease. METHODS: Since March 1998, an Audio-Video PC-based system that uses integrated services digital network (ISDN) at a bandwith of 384 Kbps, was installed at Pediatric Cardiac Surgery Department of Giovanni XXIII Hospital, Bari, Italy. On July 1999, thanks to Research Funds of the Health Ministry of Italy, similar audio-video systems that use ISDN were installed in the main pediatric institutions of provinces of Puglia and linked in a wide area network. RESULTS: We report the experience of Telemedicine of a Department of Pediatric Cardiology and Cardiac Surgery, and we try to analyze its impact on improvement of quality of care, once employed in provincial field. PMID- 11424826 TI - [Puntino and the injections of Popeye]. AB - Subcutaneous injection of active principles must be performed through a short and thin needle and an insuline syringe (because of the few quantity of drug to administrate). In our Centre, to prevent preterm chronic anemia wc practice subcutaneous therapy with recombinant human erythropoietin. 300 UI three times a week, to all the newborns weighing < 1500 g at birth. Injections to the newborns are performed in correspondence of their gluteal and deltoid muscles, and in the outer part of their thigh. To prevent atrophy, it is important to change every time the site of Injection. For this goal, we have created the shape of a newborn, nained Puntino, and we have located 24 points on it. Each point has received a number between 0 and 23. During the treatment we have followed the guide of Puntino to locate each time the correct site of injection. Thanks to Puntino's aid, there were no cases of skin adverse reaction and atrophy, even in newborns weighing < 1000 g. PMID- 11424827 TI - [Neonatal resuscitation: a fascinating story!]. AB - Over the past thousands of years many examples can be found of the treatment given for asphyxia in the apparently dead subject. The resuscitation procedures were applied alike both in adults and in newborn babies. The first reference made to the mouth-to-mouth method was in the Kings'. In the second half of the 1800 many Authors proposed a method to resuscitate the asphyxiated newborn infant, which were still practiced up until the 1930 s. Many other different methods of air and 02 administration to the airway and stomach of an apparently dead newborn were well known and used in the 1800 s and still used in the 1950 s. Other methods used involved the dilatation of the rectum; the immersion in alternating warm and cold water and the use of a larynx tube developed by Chaussier which was then modified by other Authors. The use of tight fitting tracheal tubes and direct tracheal 02 administration after the clearing of the oropharynx using fingers and finally the mouth to mask and mouth to endotracheal tube breathing. All these examples are indicative of the approach taken in the resuscitation of the apparently dead infant. The science of resuscitation and its history continue to evolve.... PMID- 11424828 TI - [Early discharge project appropriate for the healthy newborn]. AB - Recently the stay in the hospital of the pueperae and of their newborns has progressively been shortened in several Countries. A lot of reports evaluating the risks of early discharges have not shown any significant differences in the hospital readmission rates between newborn early discharged and case-controls. This phenomenon is occurring also in Italy and the early discharge after the delivery is one of the aims of the "Percorso-Nascita" in the National Health Program of 1998-2000. Since three years ago, in the Nursery of the Department of Neonatal Medicine of University of Parma we have been discharging the newborns at term in the second day of life, choosing the healthy newborns in agreement with the obstetricians. The increment of the early discharges has happened in our Department after realizing that those did not create an increment of the hospital admissions in the neonatal period. Since January 2000 in Parma the project of appropriate discharge named "around the cradle" derived from the cooperation between hospitals and territory is working. This project consists of a continuous essistential support for mothers and children throughout an organic integration between hospital and territory. The women that take part to this project have the opportunity to be discharged from the hospital between the 44 and the 56 hours after the physiological delivery, being followed at home by the local obstetrician and paediatrician. The project is divided into three parts. The first part, pre-partum, is based on the information that is delivered to all the pregnant women that are eligible; the second part, the hospital phase, implies the usual and routinary care to the pueperae and their newborns as well as the clinical neonatal and obstetrical evaluation that allows to select our cohort. In the latter phase, extra-hospital, the local paediatrician take care of the newborn and start examining him/her within 72 hours after the discharge, while the home assistance of the local obstetrician will be within 48 hours. After initial difficulties, the organization of this project is quite efficient with a constant increment of attending requests to our project by the patients. Of course this is an organizative model quite complex that needs to verify process, efficiency and results, but anyway it sees the cooperation among many professionists of different specialities: neonatologists, paediatricians, obstetricians, practitioners, all working together for a common programme. PMID- 11424829 TI - [Early discharge and breast-feeding]. AB - Early discharge is often correlated with negative events such as feeding problems leading to hypernatremic dehydration and malnutrition and non initiation or premature cessation of breastfeeding. In the reality of Parma we have valued the impact of early discharge on breastfeeding, analyzing a cohort of 267 dyads, 134 of which were in the "late discharge" group and 133 in the "early discharge" group. Our study showed, according to the most recent literature, that early postpartum discharge did not negatively affect the start and length of breastfeeding: early discharge indeed, if associated with suppression of "hospital routines" and with home-based postpartum follow-up visits, is a determining factor for the success of breastfeeding. PMID- 11424830 TI - [Evaluation and impact on incidence of admission to the neonatal care unit in cases of jaundice after early discharge form the nursery]. AB - We have evaluated the entity of admissions to the Neonatal Care Unit (NCU) for jaundice in the period 1996-2000 of babies early discharged from the Nursery (36 48 hours for spontaneous deliveries and 72-96 hours for cesarean section). We have found an increasing trend of admissions for jaundice (R2 = 0.76), proportional to the increasing practice of early discharge and the early discharge also for infants 34-37 weeks' gestation. The admissions for jaundice from home have increased about 8 times from 1996 to 2000, respectively from 8 to 65 cases. The mean value of maximum total bilirubinemia at admission was 20.5 +/- 2.7 mg/dl. None of the infants underwent exchange transfusion. The most part of the jaundices (69/123) were idiopathic in term infants, followed by those in preterm babies (24/123) and by those in ABO incompatibility without hemolytic disease (15/123). Mean age at admission of the total population was 6.5 +/- 3.0 days, and 9 newborns had signs of dehydration at admission, with weight loss between 11 and 13.8% from birth. Also the admissions for jaundice of extracomunitary babies have resulted increased and they had a longer hospitalization than the rest of the population (p = 0.02). CONCLUSIONS: Early discharge from the Nursery has determined an increase of admissions to NCU for jaundice. New criteria have to be considered to select more carefully the infants that can benefit of this practice. PMID- 11424831 TI - [Early protected discharge of the mother and the neonate: case-control study]. AB - Aim of this study is to compare traditional post-partum hospital stay to hospitalization associated with early protected discharge: a case-control study has been performed to evaluate outcome as mother's appreciation of the experience as well as breastfeeding. The study included 50 healthy-term newborns and their mothers, discharged within 24 and 48 hours of life, and 44 controls, who had traditional "rooming-in" stay, delivered at the Department of Neonatology- University of Turin. The protocol included a midwife daily home visits and a neonatologist and nurse visit within 4th to 5th day of life, to evaluate mother's and baby's health status and to perform metabolic screenings. An ambulatorial follow-up visit at 1 month of life and 2 telephone interviews, at 3rd and 6th month, were also planned. During the first week of life 45 (90%) early discharged newborns had complete nursing (breastfeeding + water or other fluids), 4 (8%) had complementary nursing (breastfeeding + formula) and 1 (2%) received formula. Among controls, 46 (92%) babies received complete nursing, 2 had complementary nursing and 2 had artificial nursing. At 6 months of life breastfeeding was complete for 2% of cases and 6% of controls; in 44% of cases and 56% of controls nursing was complementary. Readmissions to our Birth Center were 2 among early discharged newborns, 1 in the control group. About project's appreciation, 96% of early discharged and 98% of control group mothers declared their availability to repeat the experience. Caring and supporting were judged adequate in 94% of both group. By adequate supporting of mother and newborn, short and traditional hospitalization are both pleasant and don't seem to present significant differences in type and length of nursing. PMID- 11424832 TI - [Childbirth preparation courses: obstetrical and neonatal evaluation]. AB - From 1997, R.A.T. (Respiratory Autogenous Training) and "Stretching" training have been performed into the Department of Obstetrics and Gynecology University of Parma, for childbirth preparation. The aim of this study is to evaluate the obstetric characteristics of these women during labor and delivery. We compared the labour and delivery characteristics of 200 women who have completed antepartum R.A.T. and stretching training with 100 matched controls who have not. Preparation is significantly related to reduction in dystocic deliveries (operative vaginal delivery and cesarean section) and emergency cesarean section. Epidural analgesia (an obstetric procedure that is not routinely offered in the department of Parma) is more frequently performed in women prepared with ante partum training. The neonatal outcome is good in all the three groups. "Prepared childbirth" courses offer measurable clinical, obstetrical and neonatal advantages and psychological support, providing a useful link between prenatal ambulatory care and hospital labor and delivery care. PMID- 11424833 TI - [Guaranteeing fetal well-being in water childbirth: is it possible?]. AB - The criteria, which a modern obstetric department is based on, are to deliver serenely and naturally according to the mother's personal exigencies and preserving the child's right to his/her own safety. The attempt to offer the woman a natural place with respect for these principles has improved the knowledge about the physiologic changes of the female organism during labour and water birth. Our experience about water birth began on 1st of July 2000, the day of the inauguration of the new birth room of the maternal-infantile department of the hospital of Lavagna. We nursed 15 women during labour and water birth, 11 were multiparas, 4 were primiparas, the average age was 31-year-old. We used the existing criteria of maternal and fetal selection for the care of physiologic water birth with a low risk. Particularly, the fetal heart rate was monitored at least for 30 minutes before the immersion into water and then at scheduled intervals during labour. To this purpose we used a cardiotocograph provided with an ultrasound probe (with high density of crystals) and with a toco (with high sensitivity), both waterproof and wireless. In our sample the episiotomy was not performed and 3rd degree lacerations did not happen. The neonatal average weight was 3100 gr for the primiparas and 3040 gr for the multiparas, respectively. The Apgar measurement was never lower than 8. The average time of labour was 6 hours for the group of the primiparas and 4.25 hours for the multiparas, respectively. In conclusion the monitoring of fetal welfare during water labour does not substantially differ from the monitoring of traditional labour, but it requires specific equipments. PMID- 11424834 TI - [Protagonists of natural delivery]. AB - In an attempt to re-humanize such natural event as giving birth, several, maternity wards have proposed alternative solutions. Since 1985 the obstetric paediatric staff in Poggibonsi (Siena) has been using a special natural (active) room to this purpose. In this report we wish to understand the motivations which lead couples to decide to give birth in the active birth room. From daily contact with persons involved in this type of birth it has been possible to reach various conclusions regarding the persons most deeply involved in the event. The need of intimacy in a unique moment such as giving birth the almost total absence of medical intervention, the possibility of immediate and continuous contact with her baby, and also the reassuring presence of her family represent the main reasons for womans' choicfe in this matter. Even through the father cannot personally participate in the emotions of pregnancy and birth, he takes part by choosing to be as close as possible to his partner and child through his continuous and attentive presence. The mid-wife, that in this situation is far from the doctor's supervision is the person preferred by the woman giving birth because she too is a woman and because of her professional capacities, and she is therefore extremely reassuring. Last, the child-nurse represent the friend who affectionately and materially supports the mother and the family, especially concerning breast-feeding and care of the newborn baby. PMID- 11424835 TI - ["Defensive medicine" in the choice of cesarean section]. AB - AIM: To evaluate the perception of "Defensive Medicine" by hospital obstetricians and the influence of this attitude on the choice of cesarean delivery. METHODS: Questionnaire sent by mail to a sample of obstetricians of general district, teaching and university hospitals in a region of southern Italy (Puglia). Doctors were selected as the head, the senior and the junior specialist of each department. Independent variables of the study were considered as demographic data of the subjects, seniority, interest in private practice, size of the hospital, background cesarean section rate, personal and site of work exposure to legal claims. Outcome measures were experience and confidence in training for operative vaginal and breech delivery, use of the partogram in labour, opinion about a trial of labour after a previous cesarean section and about section on request, personal perception of Defensive Medicine. Univariate and multivariate analysis of data were performed. RESULTS: The response rate was 83%. According to our data, seniority in service meant confidence in obstetrics manoeuvres and more will to teach them; size of hospitals was positively related to a more rationale approach of the diagnosis of dystocia; heads of units were more prone to accept the patient's wish for a cesarean section. Doctors with large private practices were less likely to be sued and the perception of legal pressure was directly related to the rate of cesarean section in each unit. CONCLUSIONS: Defensive Medicine is a reality that encompasses all categories of doctor in this survey. The only differences were in the rate of perception of legal pressure. We believe that residential programs should be modified in order to improve specialists' understanding of malpractice problems and that the patient-doctor relationship should be ameliorated in public hospitals. PMID- 11424836 TI - [Psychosocial intervention model of neonatal intensive care in Modena]. AB - Many authors agree that psychosocial factors influence the psychophysical development of the infant. These factors must be taken into account in the neonatal care. At the neonatal intensive care unit (NICU) of the University Hospital of Modena, physicians, psychologist and nurses cooperate to take care of each newborn with a difficult psychosocial background. Together they individuate and select newborns with psychosocial difficulties, plan an integrated action and, eventually, address the family to the community services. We describe our experience with 106 cases from 1996 to 1999. The integrated action included observation of parental behavior within the NICU and psychological support for parents by means of psychological counselling. Many meetings among physicians, nurses, psychologist and the social worker were needed to achieve good results. The aim of the project was to define a method to provide individualized actions in order to favour psychophysical health of newborns and their families. PMID- 11424837 TI - [Model of integrated hospital project for the management of patients at risk of sudden death syndrome]. AB - Home cardiorespiratory monitoring (HM) is an accepted practice in infants at high risk for Sudden Infant Death Syndrome (SIDS) as those with the following conditions: 1) Siblings of SIDS; 2) Apparent Life Threatening Events (ALTE); 3) Apneas of prematurity. From 1998 the Division of Neanotology of the Policlinico of Modena has followed prospectively infants at high risk for SIDS, in collaboration with the General Health Service. To support the clinical trial 10 apnea monitors were provided by the General Health Service and managed from the Department of Neonatology. During 18 months 24 infants have been followed by HM, and among then 20 were at risk of SIDS (11 female and 9 male): 7 were siblings of SIDS (35%); 10 had previous episodes of ALTE (50%); 3 were born prematurely and had persistent apneas before discharge (15%). The mean period of HM was 5.85 months. None of the infants died for SIDS or had episodes of ALTE that required resuscitation measures. In only one case parents stopped earlier than recommended, but personal interview to parents showed that all the others families gained benefits and reassurance from HM. PMID- 11424838 TI - [It is not so difficult to carry out simple things properly!]. AB - This report is about the results of the experience we did, since 1996, in the nursing unit of a first level mountain hospital in which we work, where we followed a new childbirth and postpartum assistance protocol aimed to humanizing birth and postpartum, promoting exclusive breastfeeding and reducing childbirth risks. The results show that, since the beginning of our experience, we had an evident reduction of preterm childbirth (newborns with less than 36 weeks of gestational age). We also had a reduction of cesarian sections and of neonatal transferrals in second level neonatal units. Furthermore, we had a progressive growth of exclusive breastfeeding. The report also shows that, since 1996, the newborns discharged from our hospital are exclusively breastfed for a time longer than before and that, at 1 year, 11 percent of our children are still breastfed. PMID- 11424839 TI - [Neonatal diabetes, heterogeneous syndrome]. AB - Neonatal diabetes (ND) is a rare condition of hyperglycaemia which occurs, generally, within the first month of life, requires insulin therapy and lasts more than two weeks. The pathogenesis is uncertain and the different clinical variants are heterogeneous; both a "permanent" and a "transient" form of ND exist. We report the clinical history of three neonates (A.L., A.B. and V.I.) in which a tout-court diagnosis of ND was done; the follow-up allowed us to reevaluate the initial diagnoses and suppose different ethiopathogenic mechanisms. Clinical observations, insulin requirement and biochemical data obtained during the years of follow-up allowed to distinguish three different situations: a "permanent" ND (A.L.), a "transient-recurrent" ND (A.B.), and a Maturity Onset Diabetes of the Young (MODY) (V.I.). PMID- 11424840 TI - [Neonatal hemochromatosis: significance of early identification]. AB - Neonatal hemochromatosis (NH) is a rare clinical-pathologic entity defined by severe neonatal liver failure (NLF) of intrauterine onset associated with extrahepatic siderosis that spares reticuloendothelial elements. Whether this entity is the result of a distinct disease process or is the pathologic end result of many other forms of NLF remains to be assessed. Death from multisystem organ failure usually occurs in the first few days or weeks of life. We report two sisters with neonatal hemochromatosis with different outcome. The first died at 21 days of life for multiorgan failure and sepsis. The second was diagnosed and treated very early by aggressive support with fresh-frozen plasma, packed red cells, platelets, coagulation factors, diuretics, and pressors, and she is well at 2-year-follow-up. The few therapeutic options are discussed. In general, few patients survived NH. Among patients treated only by medical approachs and that did not undergo orthotopic liver transplantation (OLTX), favorable outcome is only very rarely reported. For management of NH infants early recognition of the disease is critical. Our experience seems to suggest that as early is the start of treatment as high is the possibility of survival. At this moment very early aggressive supportive treatment should appear to be the most useful approach to stabilize the patient hoping in spontaneous liver recovery, or, in turn, to permit OLTX before septic complications bring to clinical decline. PMID- 11424841 TI - [Steinert's myotonic dystrophy: severe neonatal form with unknown family history]. AB - A child with severe generalized hypotonia and respiratory insufficiency, with an unknown positive family history for Steinert's disease, is referred. We want to point out the importance of correct anamnesis and of physical examination during pregnancy to suspect and diagnose rare and incurable fetal pathologies. CASE REPORT: The child was born after caesarean section. Polyhydramnios and decreased fetal movements were noticed during pregnancy. At birth, the baby presented asphyxia (Apgar 4/6) and respiratory insufficiency: he was then intubed. He received assisted ventilation for 37 days. At the physical examination, the child appeared hypotonic, hyporeflexic, without sucking reflex, with arthrogryposis and ligament laxity. On first day, chest X-ray showed paralysis of the right hemidiaphragm. His mother presented with hypotonia of the facial muscles, lid drop, light muscular weakness, positivity to neostigmine test: we then assumed that the baby was affected by transient neonatal myasthenia gravis and neostigmine was carried on. Anyway, the general conditions of the baby didn't improve. We were able to establish diagnosis of Steinert's disease (a form of muscular dystrophy with autosomal dominant inheritance with incomplete penetrance) after some other examinations (negativity of acetylcholine receptor antibodies, elevation of creatine kinase level, myopathic pattern on electromyography). Gene DMPK alteration was documented with a molecular genetic test. PMID- 11424842 TI - [A neonatal case of congenital myotonic dystrophy]. AB - Congenital myotonic dystrophy is a rare autosomal disease, caused by an increased number of cytosine-thymine-guanine (CTG) trinucleotide on chromosome 19q. In the neonatal period the most peculiar clinical features are arthrogryposis, hypotonia, facial diplegia, respiratory and feeding difficulties. Clinical and electrical myotonic discharges are difficult to elicit in the newborn. We report a case of congenital myotonic dystrophy in a female newly born presenting with hypotonia, diaphragmatic paralysis, facial diplegia, and contractures of hips, knees and ankles. The diagnosis was confirmed by genetical study on lymphocyte DNA. PMID- 11424843 TI - [A case of female pseudohermaphroditism caused by maternal androluteoma]. AB - Female pseudohermaphroditism is a condition characterized by various degree of external genitalia virilization in a patient with female internal genitalia and karyotype (XX). External genitalia is masculinized congenitally when female fetus is exposed to excess androgenic environment. Fetal metabolic abnormalities, like congenital adrenal hyperplasia, are the most common causes of female pseudohermaphroditism, however there is a low incidence of gestational hyperandrogenism caused by maternal pathology. We report a case of female pseudohermaphroditism secondary to a maternal ovarian luteoma of pregnancy producing androgenic hormones. The newborn presented a severe degree of external genitalia virilization with high urogenital sinus (stage Prader V). Moreover we describe the main steps of diagnostic iter that are necessary both to exclude other causes of virilization and to study all anatomical aspects in view of the surgical correction. The operation consists in two phases of action: an early clitorisvulvoplasty according to Passerini-Glazel and a late vaginal pull-through with anterior saggital transanorectal approach (ASTRA). PMID- 11424844 TI - [Neonatal pseudohypoaldosteronism: when a denied truth can delay a diagnosis]. AB - A child with neonatal pseudohypoaldosteronism is referred. The diagnosis was delayed and complicated as her parents didn't tell us that her sister had been affected by the same illness. The child was born after premature rupture of membranes at 34 weeks. At birth, her conditions were satisfactory. The general conditions of the child gradually worsened and, though she regularly ate human pastorized bank milk, she didn't gain any weight. After 13 days she appeared dehydrated with marbled dry skin and haloed eyes, hypotonic and hyporeflexic. Her suction became more and more weaker, with frequent regurgitations and vomiting. Laboratory tests pointed out hyponatremia (110 mEq/l) and hyperkalemia (6.8 mEq/l). We were able to establish diagnosis of primary pseudohypoaldosteronism measuring plasma concentrations of aldosterone (> 5000 pg/ml), 17 hydroxyprogesterone (normal) and corticosteroids (normal). The child was then supplemented with sodium chlorure. A gradual improvement of general conditions took place. We assumed that both childrens and mother were affected by a milder form of primary pseudohypoaldosteronism, characterized by an autosomal dominant way of transmission. We found high levels of aldosterone with normal levels of blood salts also in the sister and mother patient's. After the neonatal period, pseudohypoaldosteronism can be only suspected in case of positive family history. PMID- 11424845 TI - [Symptomatic congenital cytomegalovirus infection: report of a clinical case with very severe onset]. AB - This document is about a serious congenital CMV case in a 36 weeks' gestation female newborn with intrauterine growth retardation delivered by cesarean section whose mother was drug addicted. At birth the newborn showed petechiae and bloody blisters all over the body, serious hepatosplenomegaly and microcephaly. Laboratory tests showed thrombocytopeny (platelets count 24.000/mm3) requiring platelets and fresh frozen plasma transfusions during stay in the hospital; echoencephalography showed brain cyst in occipital area, dilated ventricles and calcification in the periventricular area; cardiac echography showed congenital cardiopathy with ventricular and atrial septal defect and patent ductus arteriosus. Urinary presence of CMV-DNA and then CMV-DNA in maternal blood and milk were found. The newborn was given 4 days of endovenous iperimmune immunoglobulin, but the treatment with ganciclovir was impossible for her serious hepatopathy. The newborn was discharged at 45 days and followed monthly with a day hospital program. Now she's 10 months old and she has serious neuromotory problems with left emiparesis and she's following a program of neuromuscular re education at our hospital. PMID- 11424846 TI - [Sepsis caused by Corynebacterium xerosis in neonatology: report of a clinical case]. AB - The pathogenicity of the nondiphtheria corynebacteria, most commonly known as coryneform bacteria in humans has been recognized in the last two decades. Corynebacterium xerosis is part of the normal flora of the skin, nasopharynx, conjunctives and it has recently been isolated from vaginal swabs. During the last few years, there has been an increased number of case reports claiming an association of C. xerosis with diseases, like septicemia, endocarditis, pleuropneumonia, peritonitis, osteomyelitis, septic arthritis, mediastinitis, meningitis, ventriculitis specially in immunocompromised patients or surgical patients. Infections due to C. xerosis have been reported rarely in newborn. We report a case of sepsis due to C. xerosis in a newborn without evident immunodeficiency. Our case further support the recognition of C. xerosis as a human pathogen and reinforces the fact that it should not be routinely considered as a contaminant. PMID- 11424847 TI - [Neonatal arteriovenous thrombosis: report of a case]. AB - Limb vascular thrombosis is a severe, rare entity, often needing invasive surgery and sometimes leading to loss of function. An infant with in utero arteiro-venous thrombosis is reported. CASE REPORT: The baby, a female infant, was born at 34 weeks from a mother affected by gestational diabetes. She was referred to Neonatal Care Unit because of mild respiratory distress syndrome. She had no malformations. Spontaneous acute ischemia of right upper limb was noticed soon after admission: the limb appeared pale, cold, atonic and areflexic; no signs were noticed on it. Axillary artery and vein thromboses were soon confirmed by ultrasonography and arteriography. Intravenous administration of recombinant tissue plasminogen activator was carried on for 36 hours and followed by heparin and antithrombin III administration. Partial revascularization was obtained after a few hours: at the beginning of the treatment the limb became cyanotic, then edematous and finally the normal colour appeared everywhere but in the hand, was it became visible after a few days, when edema was reabsorbed. No recurrence of thrombosis occurred with heparin prophylaxis. Pain was treated with analgesic therapy, and motor impairment required physiotherapy. The doppler ultrasonographies which were performed subsequently confirmed a complete revascularization of axillary artery and vein. Coagulation and metabolic parameters (clotting tests, protein C, protein S, prothrombin, aminoacids) were studied but disorders predisposing to thrombosis were not found both in the patient and in her parents. CONCLUSION: Early identification of limb vascular thrombosis through external symptoms is very important, as it can avoid invasive surgical procedures. PMID- 11424848 TI - [Neonatal cystic fibrosis: report of 2 cases]. AB - Cystic fibrosis is the most common life-limiting recessive genetic disorder in Caucasian. It is caused by mutations of CFTR gene (cystic fibrosis transmembrane conductance regulator); at present over 500 mutations are known. Cystic fibrosis as a cause of respiratory distress in the neonate is quite rare. In neonatal period the most important clinical manifestations are meconium ileum and much rarely cholestatic jaundice. We present two cases of cystic fibrosis in newborns. In the first one, we point out the strict association between meconium ileum and cystic fibrosis. The patient underwent a surgical treatment for meconium ileum and the diagnosis was rapidly confirmed by genetic analysis and sweat test. The second one had intestinal obstruction from birth caused by meconium ileum associated with ileal atresia; besides, he developed cholestatic jaundice, severe and rapidly progressive respiratory disease. He died at 102 degrees day of age for cardiac failure. The diagnosis of cystic fibrosis, supported by typical clinical features and high level of serum trypsin, unfortunately wasn't confirmed by genetic analysis (lambda F508/neg), in addition, the sweat test wasn't reliable because an inadequate quantity of sweat was collected. PMID- 11424849 TI - A pediatric report card for vaccine delivery. PMID- 11424850 TI - Calling all champions. PMID- 11424851 TI - Improving immunization rates in pediatric practice. PMID- 11424852 TI - Routine childhood vaccination update: educating the office staff. PMID- 11424853 TI - The coming of age of adolescent immunization. PMID- 11424854 TI - Recommendations for storing and handling vaccines. PMID- 11424855 TI - An update on the new pneumococcal conjugate vaccine. PMID- 11424856 TI - Pediatric influenza immunization: should healthy children be vaccinated? PMID- 11424857 TI - Meningococcal disease and vaccines. PMID- 11424858 TI - A review of varicella vaccine. PMID- 11424859 TI - Mseleni joint disease progress. PMID- 11424860 TI - Endoscopic treatment of oesophageal varices. AB - Major variceal bleeding is a life-threatening complication of portal hypertension. Therapy for bleeding may be difficult and requires expertise and appropriate facilities. Endoscopic therapy using either injection sclerotherapy or band ligation after adequate resuscitation and diagnostic endoscopy is the preferred first-line treatment. Bleeding not controlled by initial endoscopic therapy requires balloon tamponade followed by repeat variceal ligation or sclerotherapy. Patients who continue to bleed after endoscopic therapy are best treated with percutaneous radiological transjugular intrahepatic portosystemic shunt stent (TIPSS) insertion. After variceal eradication, patients require lifelong surveillance endoscopy and re-obliteration of varices by endoscopic therapy if they recur. Beta-blockers to prevent recurrent bleeding are reserved for selected patients. Patients with severe liver decompensation have a poor prognosis and should be evaluated for liver transplantation. Prophylactic endoscopic therapy in patients who have never bled from varices is contraindicated as it is associated with increased morbidity and mortality. PMID- 11424861 TI - History of liver transplantation at the University of Cape Town and Groote Schuur Hospital. AB - The liver transplant programme at Groote Schuur Hospital was born out of a rich history of experimental liver transplantation and an active liver clinic. Professor John Terblanche has played a key role in the initiation and success of the programme. In the first 10 years since October 1988, 41 adults have undergone liver transplantation. The cumulative graft survival rates at 1 and 5 years were 74% and 62% respectively. PMID- 11424862 TI - Liver transplantation in children--the Red Cross War Memorial Children's Hospital experience. AB - Liver transplantation for infants and children has been available in South Africa for more than a decade. Current concerns have shifted from an initial target of early post-transplant survival to quality of life in the long term. Since 1985, 175 infants and children have been assessed, with 104 accepted for transplantation. Fifty have had orthotopic liver transplants (OLTx), 48 (3 retransplants) in Cape Town and 2 abroad. Biliary atresia was the most frequent diagnosis (52%) followed by acute liver failure (ALF) (16%). Waiting list mortality has remained high (15%), particularly for the ALF group (50%). Thirty four patients survive 1 month--14 years post-transplant. Early post OLTx mortality was low (7%) but late morbidity and mortality (23%) were mainly due to viral infection: de novo hepatitis B (5 patients, 2 deaths), EBV-related post transplantation lymphoproliferative disease (PTLPD) (6 patients, 4 deaths) and CMV disease (9 patients, 4 deaths). Tuberculosis prophylaxis, required in 6 cases, resulted in major morbidity in 1. Hypertension requiring medication along with some compromise of renal function has been present in all but 2 patients. However, all those of school-going age (20) attend school normally and remain in good health and only 3 of the survivors have abnormal liver function tests. Successful liver transplantation is possible in a developing country with limited resources. Scarcity of virus-free donors (HBV and HIV) leading to waiting list mortality and infrequent retransplantation along with long-term consequences of immunosuppression (infection, lymphoma and renal toxicity) remain problems. PMID- 11424863 TI - Gallstone pancreatitis--a critical review of current treatment strategies. AB - This review critically analyses the role of urgent biliary decompression in the management of acute gallstone pancreatitis with the emphasis on endoscopic retrograde cholangiopancreatography (ERCP) and endoscopic sphincterotomy (ES). In an analysis of currently available prospective randomised trials the role of routine urgent ERCP with or without ES in patients with predicted severe pancreatitis is challenged. Patients with persistent biliary obstruction and associated cholangitis appear to be the only subgroup who may benefit from urgent biliary intervention. PMID- 11424864 TI - Complaint rates are rising. PMID- 11424865 TI - Kernicterus threatens healthy newborns. PMID- 11424866 TI - Taking a look at the staffing assessment model. PMID- 11424867 TI - Dementia requirements no longer separated in LTC. PMID- 11424868 TI - Authorizing medical/surgical restraint protocols in hospitals. PMID- 11424869 TI - Responding to Sentinel Event Alert safety suggestions. PMID- 11424870 TI - Mast cells in allergy and host defense. AB - Mast cells have been implicated in the pathogenesis of allergic diseases and in inflammatory responses associated with pathological immune and disease-related processes including fibrosis, autoimmune pathology, and neoplasia. Recent findings in animal models of bacterial infection also suggest that mast cells may have a protective role in host defense against pathogens in innate immunity along with the probable role of mast cells in acquired immunity against parasitic infections. Mast cells are strategically located at the host-environment interface and may provide an early defense against an invading pathogen. Mast cells express an array of adhesion and immune receptors that may assist in the recognition of invading pathogens. When activated, these cells then synthesize and release key immunoregulatory cytokines, one consequence of which is to mobilize a rapid and vigorous inflammatory response. However, although it has been demonstrated that mast cells may have a role in innate immunity in defined in vitro and animal models, it remains to be determined whether mast cells are protective in innate immune responses in humans. PMID- 11424871 TI - Conjunctival mast cells in ocular allergic disease. AB - Allergic eye disease is a common clinical problem adversely affecting the quality of life for millions of sufferers. This ocular process is associated with IgE mediated conjunctival inflammation leading to signs of immediate hypersensitivity including redness, itching, and tearing. Pathologic studies have shown that the conjunctiva contains mast cells that when sensitized with IgE antibody and exposed to environmental allergens can release mediators of allergic inflammation. The type, release kinetics, and concentration of these mediators in the conjunctiva have not been completely characterized. The ability to isolate and purify mast cells and epithelial cells from human conjunctival tissue has permitted the study of mediator release and cell-to-cell signaling in this tissue. Our laboratory has developed in vitro and in vivo models to better understand how inflammatory cells are recruited to and infiltrate conjunctival tissues. These models demonstrate that mast cell activation may supply sufficient cytokine signaling to initiate and direct the well-orchestrated trafficking of eosinophils to the ocular surface, facilitate their adhesion, and cause release of potent mediators of ocular inflammation. PMID- 11424872 TI - Nasal IL-16 and MIP-1 alpha in late-phase allergic response. AB - Late-phase response in allergic rhinitis is characterized by tissue eosinophilia and influx of CD4+ T-cells. IL-16 and MIP-1 alpha are highly chemotactic on T cells and on eosinophils. Both IL-16 and MIP-1 alpha have been demonstrated to be up-regulated after challenge in the late-phase response in various atopic conditions other than allergic rhinitis. The aim of our study was to determine the expression of IL-16 and MIP-1 alpha in nasal secretions following allergen challenge in allergic rhinitis, and to compare these with characteristic late phase mediators such as IL-5 and ECP. Nasal secretions of 14 allergic volunteers challenged intranasally by their specific allergen were studied from 20 minutes to 8 hours after allergen challenge. Nasal secretions were analyzed by routine ELISA for IL-16, MIP-1 alpha, IL-5, and ECP. IL-16 and MIP-1 alpha increased significantly in nasal secretions of challenged allergic patients in the late phase response. IL-16 revealed highest amounts 5 hours after challenge, whereas MIP-1 alpha peaked at 7 hours. Both correlated significantly (r = 0.917, p < 0.05) at 6 hours. IL-5 and ECP peaked between 6 and 8 hours and correlated significantly (r = 0.951, p < 0.01) at 6 hours as well. Our data demonstrate that IL-16 and MIP-1 alpha are expressed in the late-phase response in allergic rhinitis in a more or less similar kinetic like IL-5 and ECP. They are suggested to be responsible for the observed influx of eosinophils (IL-5, IL-16, and MIP-1 alpha) and CD4+ T-cells (IL-16 and MIP-1 alpha) into the challenged allergic mucosa. PMID- 11424873 TI - MCP-1 and RANTES are mediators of acute and chronic inflammation. AB - Regulation of leukocyte migration and activation by chemokines are recognized as potentially important functions in the induction of acute and chronic inflammatory reactions. Regulated upon activation normal T cell expressed and presumably secreted (RANTES), monocyte chemotactic protein-1 (MCP-1), and related molecules constitute the C-C class of the beta chemokine supergene family with inflammatory properties. Here we report that in experimental studies RANTES and MCP-1 provoke mast cell activation and increase histidine decarboxylase mRNA expression in a dose-dependent manner. Moreover, injections of RANTES and MCP-1 in the rat skin cause mast cell, eosinophil, and macrophage recruitment, and prostaglandin E2 (PGE2) generation. In a chronic inflammatory model MCP-1 was found to mediate the recruitment of mononuclear cells in calcified granulomas. In addition, MCP-1 mediated parasitic infections caused by Trichinella spiralis. In accordance with other studies, RANTES and MCP-1 were found to play an important role in the lung allergic inflammation, lung leukocyte infiltration, bronchial hyperresponsiveness, and the recruitment of eosinophils in the pathogenesis of asthma. Here for the first time we propose a new mechanism of pulmonary airway inflammation where RANTES and MCP-1 are deeply involved. We also studied the apparent role played by RANTES in the pathogenesis of relapsing-remitting multiple sclerosis enhancing the inflammatory response within the nervous system. PMID- 11424874 TI - The chemokine network in eosinophil activation. AB - Eosinophils are predominant effector cells in allergic and autoimmune diseases attracted by several chemokines into the inflammatory tissue. In the past many new biologically active chemokines have been cloned exploring the gnomic DNA sequence data base in the vicinity of already-known chemokine sequences. This article will help to better understand the chemokine network in eosinophil activation according to the new nomenclature of chemokines. Moreover, possible new therapies, such as chemokine receptor antagonists, for the treatment of allergies are discussed. PMID- 11424875 TI - An updated model of clinical asthma. AB - A model of asthma, based on the Expert Panel Report but updated to allow for the independence of bronchial hyperresponsiveness from airway inflammation, is presented. The updated model of asthma can be summarized: (1) AI = f1(Allergen) f2(AI); (2) BHR = f3(Allergen)-f4(BHR); (3) PR = f5(AI); (4) Asthma = f6(AI + BHR + PR); where AI is airway inflammation, Allergen is sensitization by allergen, BHR is bronchial hyperreactivity, and PR is pulmonary remodeling. The updated model demonstrates that preventing bronchial hyperreactivity will not prevent the lung destruction associated with asthmatic pulmonary remodeling. The updated model of clinical asthma presented here helps to provide a basis for computer simulation of chronic asthma. PMID- 11424876 TI - The linear relationship between changes in childhood growth velocity and topical glucocorticoid dose. AB - The concept of therapeutic indices for glucocorticoid treatment of rhinitis and asthma requires demonstration of the dose dependency of benefits and side effects. Therefore, we examined the relationship between glucocorticoid dose and changes in growth velocity (delta GV). The literature was reviewed for articles where the net delta GV could be calculated among steroid and parallel placebo, active treatment, or baseline run-in periods. Steroid dose and delta GV were analyzed by linear regression for 5 rhinitis and 19 asthma studies using topical budesonide, beclomethasone, fluticasone and mometasone, parenteral steroids, and nonsteroid comparitors. Dose dependency was established between 0 and the equivalent of 2000 micrograms/day of beclomethasone (r2 = 0.60). delta GV was not significantly affected by 200 micrograms/day of BDP or less. Nasal and bronchial administrations appeared to give equivalent responses. Growth suppression occurred within 2 weeks, and may be linked to a delay in the onset of puberty. The physiology of these effects was discussed. PMID- 11424877 TI - Clinical relevance of growth studies. PMID- 11424878 TI - Clinical and basic science research on allergic rhinitis and asthma from Nassau University Medical Center. AB - Allergic inflammatory disorders involved with the upper and lower airway are related to allergic rhinitis and asthma. The role of nasal provocation and airway epithelium as models to evaluate allergic rhinitis and asthma are illustrated in studies from our research center. New concepts related to the anti-inflammatory effect of nasal steroids, antihistamines, isomers of beta 2 agonists, cromolyn sodium, and enoxaparin sodium related to airway remodeling are addressed in this article. PMID- 11424879 TI - Alternative routes of immunization for prevention of infectious diseases: a new paradigm for the 21st century. AB - The prevention of infectious diseases by the use of vaccines represents one of medicine's greatest triumphs during the 20th century. This era has witnessed the global eradication of smallpox as a result of Jennerian cowpox vaccination, the elimination of paralytic poliomyelitis from the western hemisphere, and within 5 10 years the anticipated eradication of poliomyelitis worldwide as a result of the poliovirus vaccines. Next slated for worldwide eradication is measles, the great killer of infants and children, which each year extracts a global mortality of one million victims. Throughout the 20th century the percutaneous (i.e., subcutaneous or intramuscular) route has almost exclusively been the preferred way to administer vaccines. However, as a result of several important scientific discoveries made during the 20th century, including new tissue-culturing techniques, the development of recombinant DNA technology, and genetic sequencing, a whole new generation of tailor-made modern vaccines has become available, including DNA vaccines and transgenic plant vaccines. Moreover, it became apparent that alternative routes of administration of vaccines, such as by aerosol immunization and transcutaneous skin patches, might be more appropriate and more effective than immunization via the parenteral route. This paper describes some of the recent advances relating to alternative methods of immunization and will focus primarily on the development and use of respiratory aerosolized vaccines. PMID- 11424880 TI - Quality of life in respiratory allergy. AB - The HRQL (Health Related Quality of Life) is considered a relevant aspect of pharmacological treatment, since it evaluates the outcomes from the patient's viewpoint. This article reviews the currently available information about HRQL as an outcome measure in Respiratory Allergy. Pertinent abstracts and articles were considered, showing strong evidence that HRQL assessment has emerged as an important parameter in both clinical practice and experimental trials. The different types of questionnaires used in HRQL assessment, the results of studies using those questionnaires and concomitant effect of pharmacologic treatments, are herein reviewed. Moreover, the impact of allergy specialists' care on patients with allergies and the new perspectives in HRQL are discussed. PMID- 11424881 TI - Health insurance companies finally take to the Web. PMID- 11424882 TI - Children's Hospital of Philadelphia lets clinicians do their own Web publishing. PMID- 11424883 TI - Web training for consumers, caregivers, and staff. PMID- 11424884 TI - Index of suspicion. Case 2. Diagnosis: hyperphosphatemic hypocalcemia. PMID- 11424885 TI - Index of suspicion. Case 3. Diagnosis: alcohol withdrawal seizures. PMID- 11424886 TI - Chemotherapy Foundation Symposium XVIII: Session on malignant melanoma. PMID- 11424887 TI - IBC's 6th Annual Conference on Angiogenesis: Novel Therapeutic Developments. PMID- 11424888 TI - Proceedings of the 8th International Respiratory Forum: Inhaled drug delivery. London, United Kingdom. PMID- 11424889 TI - The influence of lung deposition on clinical response. AB - Delivery of more drug to the lung may appear to be a desirable goal in the treatment of asthma and chronic obstructive pulmonary disease, since only 10 to 15% of a drug dose administered via a metered dose inhaler (MDI) reaches the lung. However, increasing the dose of most inhaled drugs may only lead to an increase in side effects, since maximal clinical benefit is usually obtained with the currently recommended dosages. Improving the regional deposition of inhaled drugs may be a more effective way of modifying clinical response. Particle size is the most significant determinant of the deposition pattern of inhaled drugs. Optimum drug delivery to the conducting airways occurs with particles ranging from 2.5 to 6 microm; particles <2.5 microm are deposited mainly in the alveoli where they may exert no pharmacodynamic effect and are rapidly absorbed, increasing the risk of systemic adverse events. Delivery devices can be compared by estimating the lung and systemic exposures, taking into account the efficacy and safety dose-response relationships for the drug-device combination. Current devices have profoundly different lung deposition profiles that could affect clinical efficacy when switching devices. Devices that achieve a high lung to systemic ratio for the inhaled drug are preferable. PMID- 11424890 TI - Why particle size should affect clinical response to inhaled therapy. AB - Studies with beta2-adrenergic agonists have shown that particle size and total dose are important determinants of optimum bronchodilation. Drug deposition in the airways is probably the most important factor for bronchodilation, since beta2-adrenoceptors and muscarinic M3 receptors are present mainly in the peripheral and central airways, respectively. Furthermore, clinical efficacy can be maintained while minimizing systemic exposure by selecting an appropriate particle size. Changes in lung function provide a means of monitoring the relationship between delivery of the bronchodilator and its efficacy, whereas there is no such immediate means of assessing antiinflammatory preventative therapy such as inhaled corticosteroids. Asthma is primarily an inflammatory disease but there are no simple tests to detect the accumulation of inflammatory cells and mediators. Data are presented to demonstrate the reduction of certain inflammatory markers in bronchial biopsy tissue taken from asthmatic patients after corticosteroid therapy. Measurement of inflammatory markers in both bronchial biopsy tissue and bronchoalveolar lavage samples may provide a way of monitoring the site of action and efficacy of inhaled corticosteroids in the future. Furthermore, it is envisaged that the effect of corticosteroid particle size on efficacy and systemic bioavailability may be investigated by exploiting these methods. PMID- 11424891 TI - Asthma inhalation delivery systems: the patient's viewpoint. AB - International guidelines for asthma management state that inhalation is the preferred route of drug delivery. Consequently, it is important to select an appropriate delivery device and to take account of a patient's preference and lifestyle, as incorrect inhaler use may lead to treatment failure. A large number of devices are available from which to select, but it may be difficult for healthcare professionals to provide guidance to the patient unless they are familiar with the characteristics and correct use of each device. Many factors affect the success of a device in a patient's hands: the device's features; the patient's psychosocial status, cultural beliefs, and language skills; and consistent instruction on inhalation technique. Constant education is essential to establish and maintain the patient's confidence in the treatment regimen. It is important that at least one member of the respiratory healthcare team should remain abreast of medical and pharmaceutical developments in the respiratory field. In this way, the choice of delivery device can be reviewed and changed to suit the changing needs of the patient. PMID- 11424892 TI - Measuring total and regional lung deposition using inhaled radiotracers. AB - The delivery of an inhaled drug to the lungs can be measured by adding a gamma emitting radiotracer to the formulation and using two-dimensional planar imaging or three-dimensional single photon emission computerized tomography (SPECT) to provide detailed information on lung deposition. The isotope most commonly used is the low energy (140 KeV) isotope, 99m technetium. Radiolabeling techniques have been successfully developed for use with nebulizers, pressurized metered dose inhalers (pMDIs), and dry powder inhaler formulations (DPI), and to investigate drug delivery to the respiratory tract for a variety of drug formulations and patient populations. However, for pMDIs and DPIs, the radiotracer is usually only physically associated with, rather than chemically bound, to the drug. Therefore, once deposited, the radiotracer may disassociate from the drug and cannot be used to track its subsequent fate; however, incorporation of a radiotracer directly into the drug molecule can overcome this. By using positron emitters such as 11carbon or 18fluorine it is possible to generate three-dimensional images of the drug in the lung using positron emission tomography (PET) scanning, which has a higher resolution and is more accurate than SPECT. Labeling drugs with PET emitters is more complex as the drug molecule must first be synthesized to contain the radioactive isotope before the drug is formulated for the inhaler. As with gamma-scintigraphy, PET scanning can be used to investigate physiological changes in the lung following therapeutic intervention, but as biological radiotracers are used, functional images (i.e., of the drug's uptake and metabolism) can also be obtained. PMID- 11424893 TI - Inhalation therapy of the future--how will it change the way we treat asthma? AB - Inhalation of drugs has made a great difference in terms of optimizing asthma therapy. The challenge for the next generation of inhalers will be to provide still greater benefits to patients in terms of efficacy, safety, and convenience. Dry powder inhalers (DPIs) are innovative new inhalation devices and comparisons with pressurized metered dose inhalers (pMDIs) show that DPIs are at least as efficient at delivering drugs, and may be a superior delivery device for some drugs. To compare different inhaler devices, drug dose comparisons that lie on the slope of the dose-response curve should be selected, and it is important to analyze the risk:benefit ratio (therapeutic ratio). Some of the new devices contain aerosols with a smaller particle size than current designs. Monodispersed aerosols with a very narrow range of particle sizes may target drug delivery to specific areas of the lung where it is most effective. However, as smaller particles are more easily absorbed into the pulmonary circulation via the alveoli, these formulations may be associated with a higher incidence of systemic side effects. The optimal particle size required to maximize the therapeutic ratio of a molecule may be different for a beta2-agonist than for an inhaled corticosteroid. A greater understanding of this relationship will be required if we are to achieve better drug targeting with future inhalers. PMID- 11424894 TI - Treating systemic diseases via the lung. AB - Inhalation of pharmacologically active substances for medicinal, social, or recreational purposes has been prevalent for centuries but experience of exploiting the lung as a route of delivery for treatment of nonrespiratory diseases is limited. Despite the success of current applications such as anesthetics, the utility of the lung for drug delivery is not well appreciated, despite advantages such as rapid onset of action. Two drawbacks are the relatively poor efficiency of current inhalation devices, especially for large molecules, and the poor patient acceptability of inhalers. Advances now being made in pulmonary delivery technology may provide the impetus needed for the development of new inhaled presentations of drugs such as peptide hormones and other biologically derived molecules. Molecules of various sizes can be delivered in clinically relevant quantities via the lung. In vitro methods show that lipophilic drugs are absorbed through the alveolar membrane more quickly. Early work in animal models has already shown that absorption of analgesic and antiinflammatory drugs that are not well absorbed orally can be improved by delivering them by inhalation. This work may soon give rise to new formulations for therapeutic use. PMID- 11424895 TI - Guidelines for devices and choices. AB - Asthma patients can be treated safely and effectively with the compounds and inhalation devices currently available. However, the choice of devices is so wide that the healthcare professional may be easily confused, and the effectiveness of treatment reduced. Clear guidelines are needed to help resolve this difficulty; however, those currently available do not contain enough useful information on the different delivery systems to assist the selection process. The function of the anti-asthma drug (i.e., preventer or reliever) should determine the choice of device. For inhaled corticosteroids, the device should provide effective and reliable delivery, while simplicity in use will aid good inhalation technique. A device that minimizes systemic absorption of these drugs will improve drug safety. The devices of choice are pressurized metered dose-inhalers (pMDIs) with spacers and some dry powder inhalers (DPIs). The main selection criteria for short-acting beta2-agonists are portability and patient acceptability. DPIs and pMDIs are the most suitable systems for these drugs. Cost is an important consideration for both drug types. PMID- 11424896 TI - Evaluation of pulmonary absorption using pharmacokinetic methods. AB - When assessing inhaled drug absorption, standard pharmacokinetic analyses cannot differentiate drug that reaches the systemic circulation from the lungs from that of the gut. Pulmonary absorption kinetics can be assessed for drugs with negligible gastrointestinal absorption or for drugs that are eliminated via first pass metabolism. For other drugs, pulmonary absorption kinetics can be studied by blocking gastrointestinal absorption with charcoal, or by studying absorption during the first 30 minutes post inhalation before appreciable oral absorption has occurred. Pharmacokinetic data generally agree well with pulmonary deposition data derived by measuring inhaled radiotracer-labelled drug using gamma scintigraphy. Monitoring pulmonary and oral absorption also provides information on total systemic exposure and therefore safety. Furthermore, the duration of action of different inhaled drugs and the degree of pulmonary targeting can be obtained by determining the corticosteroid occupancy of lung and systemic glucocorticoid receptors. Pharmacokinetic data from healthy volunteers cannot be extrapolated to patients with asthma, as systemic exposure to inhaled drugs can be markedly less in patients than in healthy volunteers. This correlates with the observed differences in side effects in these two groups. Although pharmacokinetic methods do not provide information on regional deposition, they do generate data on systemic and pulmonary exposure, and so play a role in the development of delivery systems. PMID- 11424897 TI - Statins: new drugs for treating osteoporosis? AB - Statins are widely used lipid-lowering drugs that reduce cholesterol synthesis by inhibiting 3-hydroxy-3-glutaryl-coenzyme A (HMG-CoA) reductase activity. They also strongly stimulate bone formation in rodents. If the drugs' potent bone building activity results directly from inhibition of HMG-CoA reductase, there should be less bone fracturing in humans who have taken statins to lower their serum cholesterol and prevent heart attacks, but the data gleaned from several databases are contradictory. According to some reports the lipid-lowering doses of oral statins increased bone mineral density and more than halved the risk of fracturing various bones, while according to others, including the very large Women's Health Initiative Observational Study (WHI-OS), the drugs did not significantly affect the fracturing risk. Such contradictory data could be due in part to one of the commonly used statins, pravastatin which only targets hepatocytes, or due to bone growth being stimulated by something other than inhibition of HMG-CoA reductase. Therefore, different doses of statins may be needed to build bone or optimally lower serum cholesterol. To be able to answer the question posed by the title of this editorial, it will be necessary to carry out a controlled trial using designer statins that are less liver-oriented and thus better for assessing the optimal doses needed, specifically for osteogenicity rather than for their cholesterol-lowering ability. PMID- 11424898 TI - Pharmacology and clinical efficacy of desloratadine as an anti-allergic and anti inflammatory drug. AB - Desloratadine is a biologically active metabolite of the second-generation antihistamine loratadine. Desloratadine is a highly selective peripheral H1 receptor antagonist that is significantly more potent than loratadine. Results of in vitro and in vivo studies have suggested that desloratadine has anti-allergic effects that are unrelated to its ability to antagonise the effects of histamine. Desloratadine inhibits the expression of cell adhesion molecules, inhibits the generation and release of inflammatory mediators and cytokines, attenuates eosinophil chemotaxis, adhesion and superoxide generation. Studies in animals indicate that desloratadine does not cross the blood-brain barrier and therefore does not cause sedation and does not impair cognition or psychomotor performance. Desloratadine has an excellent overall safety profile. It has no effect on QRS and QTc intervals and does not cause arrhythmias. Desloratadine is not associated with any significant changes in gastrointestinal function. In clinical studies, oral desloratadine is rapidly absorbed and bioavailability is not affected by ingestion with food or grapefruit juice. The half-life of desloratadine in humans is 27 h; the linear kinetic profile is unaltered by race or gender. Desloratadine is not a substrate for P-glycoprotein or organic anion transport polypeptide and the drug does not appear to be metabolised to a significant extent by the cytochrome P450 CYP3A4 pathway. It therefore may be safely administered with ketoconazole, erythromycin, fluoxetine, or azithromycin. Clinically, desloratadine effectively controls both nasal and non-nasal symptoms of seasonal allergic rhinitis (SAR), including nasal congestion. Desloratadine also provides significant relief of SAR symptoms in patients with co-existing asthma and is effective in the treatment of chronic idiopathic urticaria. Desloratadine improves quality of life and is well-tolerated. PMID- 11424899 TI - 11th International Congress of Endocrinology. AB - The Olympics of endocrinology, the 11th International Congress of Endocrinolgy was held rather appropriately in Sydney, four weeks after the summer games of the XXWIIth Modern Olympiad. Both occasions were a great success and whilst it may be tempting to extend the analogy to the pool or the track or heaven forbid, digress into 'drugs in sport', this review will focus on endocrinology. There were over 3000 participants with ten plenary lectures, 20 meet-the-expert sessions, 41 symposia, 128 oral free communications and 1500 posters. Sydney post-Olympics provided a vibrant, exciting and picturesque setting with outstanding convention facilities. The Congress Party was held at Campbells Cove in the lee of the Harbour Bridge looking toward the Opera House which provided an opportunity for delegates to view the two architectural icons that had become so familiar in the preceding months. Credit must be given both to the Local Organising Committee of Sydney endocrinologists who made it all happen and to the International Program Organising Committee who crafted a pageant of first rate endocrinology. It is self-evident that this report can only hope to give the reader a flavour of a Congress such as this with the choice of topics being largely idiosyncratic. With five concurrent symposia and two concurrent orals each morning and afternoon of the four days, any omissions reflect not on the topic or its importance but on this reviewer's inability to be in more than one place at once! PMID- 11424900 TI - Bibliography of toxinology. PMID- 11424901 TI - ALT-711 decreases cardiovascular stiffness and has potential in diabetes, hypertension and heart failure. AB - Vascular and/or myocardial stiffness is a major problem in ageing, diabetes, hypertension and heart failure. The development of the stiffness is partly due to the formation of glucose-dependent cross-links in the collagen. ALT-711 cleaves these cross-links. In aged-rhesus monkeys, ALT-711 decreases vascular stiffness and this effect is reversible. ALT-711 also decreases myocardial stiffness in the monkeys but this effect is not reversible in 39 weeks. ALT-711 has potential in the treatment of the stiffness associated with diabetes, hypertension and heart failure. PMID- 11424902 TI - Neurotoxicity as a mechanism for neurodegenerative disorders: basic and clinical aspects. AB - This three day meeting focused on chronic neurodegenerative diseases such as Parkinson's disease (PD), Alzheimer's disease (AD), and amylotrophic lateral sclerosis (ALS). It attracted 69 participants from 10 countries with dominance of Chile and USA. Neurodegeneration and its prevention increasingly gain in importance as the number of people affected increases year-by-year. The meeting addressed various basic aspects having pragmatic implications such as: oxidative stress, inflammatory reaction, glial activation, role of glutamatergic system and apoptosis using a plethora of in vitro and in vivo methods. PMID- 11424903 TI - The phototropin family of photoreceptors. PMID- 11424904 TI - Biochips to Bavarian cows. PMID- 11424905 TI - Medicinal biotechnology products: big problems or big opportunities? PMID- 11424907 TI - Bibliography. Current world literature. Demyelinating diseases. PMID- 11424906 TI - Computational cell biologists snowed in at Cranwell. AB - Cell biology is being inundated by an avalanche of data from the genomics and proteomics enterprises. The complexity and sheer volume of information threaten to overwhelm the ability of traditional cell biologists to grasp its implications and develop experimentally testable hypotheses. For this reason, some have begun to explore computational approaches towards organizing complex data into quantitative models. This requires communication and collaboration between the biological science community and and the physical and mathematical sciences communities. A recent meeting [The First International Symposium on Computational Cell Biology, Cranwell Resort, Lenox, MA, USA; 4-6 March 2001. Organizers: J.H. Carson, A. Cowan, and L.M. Loew (www.nrcam.uchc.edu/conference).] made a first attempt to bring these two communities together. Three feet of new snow fell during the meeting, but the 125 attendees, an unusual mixture of cell biologists, computer scientists, mathematicians, physicists, and engineers, were having too much fun defining the new field of computational cell biology to notice that they were literally snowed in. PMID- 11424908 TI - Bibliography. Current world literature. Headache. PMID- 11424909 TI - Bibliography. Current world literature. Inflammatory diseases. PMID- 11424910 TI - Index of suspicion. Case 2. Diagnosis: hyperglycemic nonketotic hypertonicity (HNKH). PMID- 11424911 TI - Index of suspicion. Case 3. Diagnosis: scurvy. PMID- 11424912 TI - ACOG committee opinion scheduled Cesarean delivery and the prevention of vertical transmission of HIV infection. Number 234, May 2000 (replaces number 219, August 1999). PMID- 11424913 TI - ACOG committee opinion. Hormone replacement therapy in women treated for endometrial cancer. Number 234, May 2000 (replaces number 126, August 1993). PMID- 11424914 TI - ACOG practice bulletin. Surgical alternatives to hysterectomy in the management of leiomyomas. Number 16, May 2000 (replaces educational bulletin number 192, May 1994). PMID- 11424915 TI - Scientists signal the way forward. PMID- 11424916 TI - On par with PPARs. PMID- 11424917 TI - Divide and rule: the bacterial chromosome. PMID- 11424918 TI - Sex and lifespan. PMID- 11424919 TI - Mutations in cis can confound genotype-phenotype correlations in hypertrophic cardiomyopathy. PMID- 11424920 TI - Identification of a large rearrangement of the BRCA1 gene using colour bar code on combed DNA in an American breast/ovarian cancer family previously studied by direct sequencing. PMID- 11424921 TI - A unique form of autosomal dominant cataract explained by gene conversion between beta-crystallin B2 and its pseudogene. PMID- 11424922 TI - Novel mutations of TMPRSS3 in four DFNB8/B10 families segregating congenital autosomal recessive deafness. PMID- 11424923 TI - Increased risk of sensorineural hearing loss and migraine in patients with a rare mitochondrial DNA variant 4336A>G in tRNAGln. PMID- 11424924 TI - Split hand/split foot malformation associated with sensorineural deafness, inner and middle ear malformation, hypodontia, congenital vertical talus, and deletion of eight microsatellite markers in 7q21.1-q21.3. PMID- 11424925 TI - A missense mutation in the SEDL gene results in delayed onset of X linked spondyloepiphyseal dysplasia in a large pedigree. PMID- 11424926 TI - Molecular characterisation of a new case of microphthalmia with linear skin defects (MLS). PMID- 11424927 TI - Cryptic telomeric rearrangements in subjects with mental retardation associated with dysmorphism and congenital malformations. PMID- 11424928 TI - Laryngeal atresia, encephalocele, and limb deformities (LEL): a possible new syndrome. PMID- 11424929 TI - Plant pathology. Reverend Berkeley's devil. PMID- 11424930 TI - Evolving issues in oncology: What is the "optimal" hemoglobin level? Symposium proceedings, part 1. September 7-9, 2000. Seville, Spain. PMID- 11424931 TI - Re: Malpractice occurrence in emergency medicine: does residency training make a difference? PMID- 11424932 TI - Re: The impact of Emergency Medicine (EM) residency training on quality of care. PMID- 11424933 TI - Re: A case of hypokalaemia-induced QT interval prolongation. PMID- 11424934 TI - Acute aortic dissection after cocaine and sildenafil abuse. PMID- 11424935 TI - [Establishment of routines for elective laparoscopic surgery]. AB - BACKGROUND: During the autumn of 1999, elective laparoscopic cholecystectomy was introduced as a new surgical procedure in the Central Hospital Ostfold, Askim. A prospective evaluation of perioperative patient logistics and patient satisfaction was performed. MATERIAL AND METHODS: 214 patients were evaluated in the ambulatory; 116 of them were scheduled for operation. The operations were performed in general anaesthesia with continuous infusion of propofol and short acting opioides. Analgetics and anti-emetics were given pre- and perioperatively as prophylaxis against postoperative pain and nausea. RESULTS: There were three unexpected cancellations. Two patients had serious postoperative bleedings and two had symptoms indicating residual bile duct stones. Twenty-eight patients (24%) were discharged on the same day. 114 patients (98%) were satisfied with their stay in hospital. INTERPRETATION: The established patient logistics proved efficient and rational. PMID- 11424936 TI - What makes catchment management groups "tick"? AB - The work of catchment management groups throughout Australia represents a significant economic and social investment in natural resource management. Institutional structures and policies, the role of on-ground coordinators, facilitation processes, citizen participation and social capital are critical factors influencing the success of catchment management groups. From a participant-researcher viewpoint, this paper signposts research directions and themes that are being pursued from the participant/coordinator, catchment group, and lead government/non-government agency perspective on the influence of these factors on the success of a catchment management group in the Pumicestone Region of Southeast Queensland, Australia. Research directions, themes and discussion/reflection points for practitioners include--the importance of understanding milieu; motivation; success; having fun; "networking networks"; involvement of "nontraditional" stakeholders; development of stakeholder/participant partnerships; learning from other practitioners; methods of stakeholder/participant representation; evaluation; the need for guiding principles or philosophy; the equivalence of planning, implementation, evaluation, and resourcing; catchments as fundamental units of Nature; continuity of support for groups; recognising a new role for government; working with existing networks; and the need for an eclectic approach to natural resource management. PMID- 11424937 TI - A conserved pseudouridine modification in eukaryotic U2 snRNA induces a change in branch-site architecture. AB - The removal of noncoding sequences (introns) from eukaryotic precursor mRNA is catalyzed by the spliceosome, a dynamic assembly involving specific and sequential RNA-RNA and RNA-protein interactions. An essential RNA-RNA pairing between the U2 small nuclear (sn)RNA and a complementary consensus sequence of the intron, called the branch site, results in positioning of the 2'OH of an unpaired intron adenosine residue to initiate nucleophilic attack in the first step of splicing. To understand the structural features that facilitate recognition and chemical activity of the branch site, duplexes representing the paired U2 snRNA and intron sequences from Saccharomyces cerevisiae were examined by solution NMR spectroscopy. Oligomers were synthesized with pseudouridine (psi) at a conserved site on the U2 snRNA strand (opposite an A-A dinucleotide on the intron strand, one of which forms the branch site) and with uridine, the unmodified analog. Data from NMR spectra of nonexchangeable protons demonstrated A-form helical backbone geometry and continuous base stacking throughout the unmodified molecule. Incorporation of psi at the conserved position, however, was accompanied by marked deviation from helical parameters and an extrahelical orientation for the unpaired adenosine. Incorporation of psi also stabilized the branch-site interaction, contributing -0.7 kcal/mol to duplex deltaG degrees 37. These findings suggest that the presence of this conserved U2 snRNA pseudouridine induces a change in the structure and stability of the branch-site sequence, and imply that the extrahelical orientation of the branch-site adenosine may facilitate recognition of this base during splicing. PMID- 11424939 TI - White dwarfs and dark matter. PMID- 11424938 TI - The mechanism and regulation of deadenylation: identification and characterization of Xenopus PARN. AB - In Xenopus oocytes, the deadenylation of a specific class of maternal mRNAs results in their translational repression. Here we report the purification, characterization, and molecular cloning of the Xenopus poly(A) ribonuclease (xPARN). xPARN copurifies with two polypeptides of 62 kDa and 74 kDa, and we provide evidence that the 62-kDa protein is a proteolytic product of the 74-kDa protein. We have isolated the full-length xPARN cDNA, which contains the tripartite exonuclease domain conserved among RNase D family members, a putative RNA recognition motif, and a domain found in minichromosome maintenance proteins. Characterization of the xPARN enzyme shows that it is a poly(A)-specific 3' exonuclease but does not require an A residue at the 3' end. However, the addition of 25 nonadenylate residues at the 3' terminus, or a 3' terminal phosphate is inhibitory. Western analysis shows that xPARN is expressed throughout early development, suggesting that it may participate in the translational silencing and destabilization of maternal mRNAs during both oocyte maturation and embryogenesis. In addition, microinjection experiments demonstrate that xPARN can be activated in the oocyte nucleus in the absence of cytoplasmic components and that nuclear export of deadenylated RNA is impeded. Based on the poly(A) binding activity of xPARN in the absence of catalysis, a model for substrate specificity is proposed. PMID- 11424940 TI - Taxing debate for taxonomists. PMID- 11424941 TI - Taxing debate for taxonomists. PMID- 11424942 TI - Taxing debate for taxonomists. PMID- 11424943 TI - Taxing debate for taxonomists. PMID- 11424944 TI - Invasive carp in the Mississippi River Basin. PMID- 11424945 TI - Clioquinol's return: cautions from Japan. PMID- 11424946 TI - Forum for agricultural biotechnology debates. PMID- 11424947 TI - Introduction to the special issue on neuroprotection by steroids: new perspectives. PMID- 11424948 TI - Steroid synthesis and metabolism in the nervous system: trophic and protective effects. AB - Steroids influence the activity and plasticity of neurons and glial cells during early development, and they continue to exert trophic and protective effects in the adult nervous system. Steroids are produced by the gonads and adrenal glands and reach the brain, the spinal cord and the peripheral nerves via the bloodstream. However, some of them, named "neurosteroids", can also be synthesized within the nervous system. They include pregnenolone, progesterone, dehydroepiandrosterone and their reduced metabolites and sulfate esters. Little is known concerning the regulation of steroid synthesis in the nervous system, which involves interactions between different cell types. For example, the synthesis of progesterone by Schwann cells in peripheral nerves is regulated by a diffusible neuronal signal. Neurotrophic and neuroprotective effects of steroids have been documented both in cell culture and in vivo. PROG plays an important role in the neurological recovery from traumatic injury of the brain and spinal cord by mechanisms involving protection from excitotoxic cell death, lipid peroxydation and the induction of specific enzymes. After transection of the rat spinal cord, PROG increases the number of nitric oxide synthase expressing astrocytes immediately above and below the lesion. PROG also plays an important role in the formation of new myelin sheaths. This has been shown in the regenerating mouse sciatic nerve after lesion and in cocultures of sensory neurons and Schwann cells. PROG promotes myelination by activating the expression of genes coding for myelin proteins. The modulation of neurostransmitter receptors, in particular the type A gamma-aminobutyric acid, the N-methyl-D aspartate and the sigma 1 receptors, is involved in the psychopharmacological effects of steroids and allows to explain their anticonvulsant, anxiolytic, antidepressive and sedative effects as well as their influence on memory. Pregnenolone sulfate has been shown to reverse age-related deficits in spatial memory performance and to have protective effects on memory in different models of amnesia. PMID- 11424949 TI - The action of steroid hormones on peripheral myelin proteins: a possible new tool for the rebuilding of myelin? AB - The present paper summarizes recent results we have obtained while studying the effect of sex steroids on the gene expression of two peripheral myelin proteins, the glycoprotein Po (Po) and the peripheral myelin protein 22 (PMP22). In particular, we have analyzed the effect of progesterone (P), testosterone (T) and their 5alpha- and 3alpha-5alpha-reduced derivatives [respectively, dihydrotestosterone (DHTT) and 5alpha-androstan-3alpha, 17beta-diol (3alpha-diol) for T, and dihydroprogesterone (DHP) and tetrahydroprogesterone (THP) for P]. The data obtained, utilizing different in vivo and in vitro experimental models, have indicated that: a) DHP is able to enhance the low messenger levels of Po present in the sciatic nerve of aged male rats; b) P, DHP and THP treatments stimulate the gene expression of Po in the sciatic nerve of adult male rats or in cultures of rat Schwann cells, while only THP is effective on PMP22; c) P and DHP are also able to increase the low messenger levels of Po present in transected sciatic nerve; d) the removal of circulating androgens by castration is able to decrease the mRNA levels of Po in the sciatic nerve, a phenomenon which is counteracted by the consequent treatment with DHT; e) the stimulatory effect of DHT on the gene expression of Po is also evident in cultures of rat Schwann cells, but in this case the effect seems to be due to the interaction of this steroid with the progesterone receptor; f) in cultures of Schwann cells PMP22 mRNA levels are stimulated only by 3alpha-diol treatment. Taken together, these observations showing the positive effects of sex steroid hormones on the gene expressions of Po and PMP22, suggest that a treatment with these molecules or their synthetic agonists may be useful in cases in which the rebuilding of myelin is necessary. PMID- 11424950 TI - Gonadal steroid enhancement of facial nerve regeneration: role of heat shock protein 70. PMID- 11424951 TI - Neuroprotective activities of estrogen: an update. AB - For many molecules, which have been found first in a certain tissue and with a certain activity in the body, multiple activities have been discovered decades later. The steroidal compound estrogen (estradiol) is certainly such a molecule. What was first described as the female sex hormone is now well acknowledged as a central neuroactive and neuromodulatory molecule. One important aspect of estrogen's effects on neurons is its neuroprotective activity. Cellular and molecular studies underline powerful neuroprotective functions of estrogen which are characterized by long-term "classical" genomic effects as well as by rapid activities depending on estrogens interaction with neuronal membranes and intracellular signal transduction pathways. In addition, estrogen has been shown to bear an intrinsic antioxidant structure that lies in the phenolic moiety of the steroidal compound. This antioxidant activity of estrogen and estrogen derivatives may provide an antioxidant "chemical shield" for neurons and may complement other neuroprotective activities of estrogen and therefore also mediate some of the beneficial effects of estrogen replacement with respect to Alzheimer's Disease. Although, investigated for decades estrogen and its receptors may still hold many surprises that remain to be identified. With the advent of novel genomic techniques, the neuronal target genes of estrogen will be identified in the future. PMID- 11424952 TI - Estrogens, apoptosis and cells of neural origin. AB - In mammals, estrogens have a multiplicity of effects in all known neural cells. We review here some of the mechanisms enabling estrogens to differentiate their influence on neural targets. In view of the potential interest in the use of estrogens in the therapy of several pathologies of the nervous system, we have proposed the use of a reductionist approach for the systematic understanding of estrogen activities in each specific type of target cell. We have therefore generated a model system in which to study the activation of one of the known estrogen receptors: estrogen receptor alpha. This system allowed us to identify a number of novel genes, the expression of which may be influenced following the activation of this receptor subtype by estradiol. We here report on preliminary data obtained by the study of one of these target genes, nip2, which encodes a proapoptotic protein product. We hypothesize that Nip2 might be an important molecular determinant for estrogen anti-apoptotic activity in cells of neural origin. PMID- 11424953 TI - The rapid effects of estrogen are implicated in estrogen-mediated neuroprotection. AB - A review of the literature reveals an impressively broad spectrum of effects of the hormone estrogen in the CNS. One of the more recently documented is the ability of estrogen to protect neurons from cell death associated with a variety of insults. While some of these actions can be attributed to nuclear effects of the hormone, mediated by the estrogen receptors alpha and/or beta, an increasing number of these effects appear to result from actions of the hormone mediated by signal transduction pathways traditionally associated with activation of membrane receptors. Here, we review current findings on actions of estrogen mediated by two pathways: those dependent on cAMP and Protein Kinase A, and those related to activation of the Mitogen Acivated Protein Kinase cascade. The evidence that estrogen can rapidly activate either pathway, and the potential involvement of the estrogen receptors alpha or beta acting in the vicinity of the cell membrane will be discussed. The possible role of MAP-Kinase activation and BCL-2 induction in the phenomenon of estrogen-neuroprotection will also be addressed. PMID- 11424954 TI - Estrogen: a multifunctional messenger to nigrostriatal dopaminergic neurons. AB - Gonadal steroids affect a wide variety of functions in the mammalian brain ranging from the regulation of neuroendocrine systems and the modulation of behavior to the stimulation of differentiation and plasticity of distinct neuronal populations and circuits. The last decades have also demonstrated that estrogen serves as a neuroprotective factor for distinct neurodegenerative disorders. Such neuroprotective effects of estrogen are most obvious for Parkinson's and Alzheimer's disease. Despite this knowledge, little is known about the mechanisms and cellular targets by that estrogen might elicit its protective influence. In the past, we have intensively studied the effects of estrogen on midbrain dopaminergic neurons which represent the most affected cell population during Parkinson's disease. These studies were mainly performed on developing dopaminergic cells and revealed that estrogen is an important regulator of plasticity and function of this neuronal phenotype. Precisely, we found that dopaminergic neurons are direct targets for estrogen and that estrogen stimulates neurite extension/branching and the expression of tyrosine hydroxylase, the key enzyme in dopamine synthesis. Together with other in vivo studies, we might draw the conclusion that estrogen is required for the plasticity and activity of the developing and adult nigrostriatal system. The presence of the estrogen-synthesizing enzyme aromatase within the nigrostriatal system further supports this idea. Surprisingly, estrogen effects on nigrostriatal cell function are not only transmitted by classical nuclear estrogen receptors but also depend on nonclassical estrogen actions mediated through putative membrane receptors coupled to diverse intracellular signaling cascades. In the future, it has to be elucidated whether nonclassical mechanisms besides genomic actions also contribute to estrogen-mediated neuroprotection in the adult CNS. PMID- 11424955 TI - Neuroprotective effects of estrogen upon the nigrostriatal dopaminergic system. AB - In this mini-review are first presented data which highlight the capacity and characteristics for estrogen to act as a neuroprotectant of the nigrostriatal dopaminergic system. Estrogen administration significantly attenuates the degree of striatal dopamine depletion to neurotoxins (MPTP, 6-OHDA and methamphetamine) which target the nigrostriatal dopaminergic system. This neuroprotection appears maximal with the 17-beta isomer, but some neuroprotection may also be present with the 17-alpha isomer of estradiol. Treatment with the anti-estrogen, tamoxifen, abolished estrogen's neuroprotective effects upon MPTP or methamphetamine-induced neurotoxicity. Estrogen also preserves striatal dopamine concentrations in gonadectomized male mice treated with MPTP, but neither testosterone nor dihydrotestosterone offer any neuroprotection. A brief review indicating a variety of potential means by which estrogen can function as a neuroprotectant is indicated. Included within this survey are presented estrogen's action as an antioxidant along with its capacity to affect monoamine oxidase, dopamine receptors/release, membrane morphology/fluidity, thermoregulation, blood flow and nitric oxide. This discussion is followed by a more detailed description of estrogen's actions upon the dopamine transporter, which is hypothesized to serve as one of the major mechanism involved with nigrostriatal dopaminergic neuroprotection. Overall, estrogen appears to inhibit dopamine transporter function by decreasing the affinity of the transporter. Such an effect could prevent neurotoxic agents from entering dopamine nerve terminals, thereby decreasing nigrostriatal neurodegeneration. Finally, some implications regarding this neuroprotectant capacity of estrogen are considered. PMID- 11424956 TI - Estradiol is a neuroprotective factor in in vivo and in vitro models of brain injury. AB - Many clinical studies suggest that estrogen enhances memory and cognition and protects against neurodegenerative diseases and injury associated with stroke or stress. These results are strongly supported by experiments performed in animal models using both in vivo and in vitro methods. We present here data from our lab that establishes that physiological levels of estradiol exert profound protective actions against ischemic injury. Further we will present evidence that these effects may be mediated through estrogen receptors that may influence the bcl-2 family of genes. PMID- 11424957 TI - Culture models for the study of estradiol-induced synaptic plasticity. AB - Estrogen, which classically affects areas of the brain related to reproduction, has also been found to affect brain regions important in learning and memory. Additionally, it has been suggested that estrogen exerts protective effects against neurodegenerative diseases such as Alzheimer's disease. Important mechanisms by which estrogen may confer protection are through the maintenance or modulation of existing synapses, or by the production of new ones. It has now been demonstrated that estrogen can increase synaptogenesis and spine production in the hippocampus, both in vivo as well as in primary hippocampal neurons in culture. The latter model system is the primary focus of this review. Synaptogenesis and spine production have been well characterized in developing and adult animals, and parallels between the synaptic morphology reflecting these processes can be readily observed in high-density primary hippocampal cultures. Moreover, in culture, estrogen induces a variety of ultrastructural modifications, many of which occur in vivo, that have been linked to various in vivo models of learning and memory. For these reasons, high-density hippocampal culture systems should be regarded as valuable tools with which to predict in vivo physiology, and may well be particularly useful for studies of the neuroprotective effects of estrogen. PMID- 11424958 TI - Estradiol attenuation of beta-amyloid-induced toxicity: a comparison o. AB - 17Beta-estradiol (betaE2) has been shown to attenuate the toxicity of beta amyloid peptides (A beta) in neuronal cultures with the effective concentration of betaE2 ranging from low nM to high microM. This study compares the effective neuroprotective concentration of betaE2 against both A beta-mediated toxicity in a human neuroblastoma cell line, SK-N-SH using cellular reduction of 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) as an endpoint to the effective betaE2 concentration obtained using a calcein acetoxymethyl ester (calcein AM) viability assay. The minimum betaE2 concentration required for protection varied 1000-fold between the two viability assays with 1 nM betaE2 conferring significant protection in the calcein AM assay but 1 microM betaE2 required for significant protection in the MTT assay Interestingly, the maximal inhibition of MTT reduction occured at sub-toxic A beta concentrations and did not correlate with other markers of cellular viability including calcein fluorescence, dye exclusion (propidium iodide or trypan blue), cellular ATP levels, or reduction of another tetrazolium dye, 5-(3-carboxymethoxyphenyl)-2 (4,5-dimethylthiazolyl)-3-(4-sulfophenyl) tetrazolium (MTS). By contrast, there was no difference between the MTT and calcein AM assays with respect to H2O2 toxicity or the neuroprotective effectiveness of 10 nM betaE2 against H2O2 toxicity. These results indicate that low concentrations of betaE2 can attenuate A beta and H2O2 toxicity in a human neuroblastoma cell line. Further, these results suggest that the MTT assay is not an appropriate assay for the determination of betaE2-mediated attenuation of A beta toxicity. PMID- 11424959 TI - Insulin-like growth factor-I receptors and estrogen receptors interact in the promotion of neuronal survival and neuroprotection. AB - Several in vitro and in vivo studies have shown that estrogen has neuroprotective properties. The neuroprotective effects of estrogen are probably exerted through several mechanisms. It is established that estrogen can provide neuroprotection by actions that are independent of estrogen receptor activation. In addition, in several experimental models, activation of estrogen receptors appears to be indispensable for neuroprotection. This review focuses on neuroprotection mediated by estrogen receptors. The interaction of estrogen with growth factor receptor signaling to induce neuroprotection is discussed. Evidence is presented that estrogen receptors and insulin-like growth factor-1 receptors interact in the promotion of neuronal survival and neuroprotection. PMID- 11424960 TI - Endangerment of the brain by glucocorticoids: experimental and clinical evidence. AB - In this paper, we review studies suggesting that elevated glucocorticoids increase the susceptibility of the brain to adverse events. The events themselves can be varied, and their effects on the brain can also differ. The common feature is that glucocorticoids may modulate the likelihood (risk) of damage or malfunction following adversity. In the first part of our paper, we describe experimental studies on the brain's cellular response to neurotoxins that support this thesis; in the second, we survey clinical evidence that indicate that glucocorticoids may endanger the brain's response to adverse social events. We suggest that there may be common features in the experimental and clinical findings. To begin, however, we draw attention to some of the properties of the hypothalamic-pituitary-adrenal (HPA) axis that seem relevant to our discussion. PMID- 11424961 TI - Near infrared spectrophotometry for the diagnosis of vasculogenic erectile dysfunction. AB - A specialized near infrared spectrophotometry instrument for noninvasive, continuous monitoring of the hemodynamic events of erection in the human penis has been developed. Its potential application for the diagnostic evaluation of erectile dysfunction was investigated. Thirty-eight patients and 18 volunteer subjects underwent penile near infrared spectrophotometry using an optical sensor probe with wavelength selectivity for hemoglobin absorption spectra. Penile blood volume changes and their time courses were measured following intracavernous pharmacostimulation in patients and visual sexual stimulation in volunteers. Spectrophotometric results were compared with results obtained simultaneously using color duplex ultrasonography, strain gauge penile circumference monitoring, penile tonometry, and clinical assessments. Spectrophotometric recordings of penile erection showed measurable blood volume changes consistent with the hemodynamic events of this biological function. Blood volume per cent (BV%) increase correlated with clinical ratings of erection quality (P < 0.001), penile rigidity measurements (P < 0.005), and penile circumference increases (P < 0.0001), and it correlated with mean peak systolic velocity measurements when BV% increase was restricted to values less than 50% (P < 0.001). The time to reach half the maximum blood volume change (BV T1) correlated directly with the time to reach half the maximum penile circumference size increase (P < 0.001), whereas BV T4 correlated inversely with mean resistive index measurements only when BV T(1/2) was restricted to values greater than 120 s (P < 0.05). Spectrophotometric criteria consisting of BV % less than 35% and BV T(1/2) greater than 120 s affirmed the diagnosis of severe erectile impairment with a similar degree of accuracy as standard ultrasonographic criteria (P < 0.002). Penile near infrared spectrophotometry is a safe, inexpensive and simply used biomedical optics technique that provides quantitative measurements of the vascular physiology of penile erection and appears to offer clinical utility in the diagnosis of vasculogenic erectile dysfunction. PMID- 11424962 TI - EP 60761- and EP 50885-induced penile erection: structure-activity studies and comparison with apomorphine, oxytocin and N-methyl-D-aspartic acid. AB - The effect of 10 peptides structurally related to the growth hormone (GH) releasing peptide hexarelin, injected into the paraventricular nucleus of the hypothalamus (PVN), on penile erection was studied in male rats. Six out of the 10 peptides tested induced penile erection in a dose-dependent manner. Among them, the most potent were EP 80661, EP 60761 and EP 91072, which were active at doses of 20-200 ng. The potency of these peptides in inducing penile erection is comparable to that of apomorphine, oxytocin and N-methyl-D-aspartic acid similarly injected into the PVN. Other peptides found active were EP 50885, EP 90101 and EP 91071, which induced penile erection at doses of 200-2000 ng. In contrast, EP 51322, EP 70555, EP 51216 and EP 91073 were inactive, as were hexarelin, EP 40904 and EP 40737 in a previous study. The majority of EP peptides found active when injected into the PVN induced penile erection, although to a lesser extent, also when given systemically (endovenously). The proerectile effect of EP peptides was prevented by the oxytocin receptor antagonist [d(CH2)5 Tyr(Me)2-Orn8]-vasotocin given into the lateral ventricles but not into the PVN, by the nitric oxide (NO) synthase inhibitor N(G)-nitro-1-arginine methyl ester given either into the lateral ventricles or into the PVN, by the N-type Ca2+ channel blocker omega-conotoxin GVIA and by morphine, but not by the dopamine receptor antagonist cis-flupenthixol or by the N-methyl-D-aspartic acid receptor antagonist dizolcipine, given into the PVN. As the structure-activity relationship of EP peptides for proerectile activity is different from those of other biological actions of these compounds, ie for GH release and eating behaviour, the present results suggest that EP peptides induce penile erection by acting on specific hypothalamic receptor sites that activate paraventricular oxytocinergic neurons projecting to extrahypothalamic brain areas that mediate this sexual function by a mechanism similar to that of dopamine receptor agonists, oxytocin and N-methyl-D-aspartic acid. PMID- 11424963 TI - Color Doppler ultrasound assessment of penile vascular system in men with Peyronie's disease. AB - The purpose of this study was to evaluate the penile vascular function by color Doppler ultrasonography (CDU) in patients with Peyronie's disease. A total of 136 men with Peyronie's disease were stratified according to their potency by history as Group I consisting of potent patients and Group II consisting of patients with erectile dysfunction. They were all evaluated by penile CDU. Overall, 70.6% had erectile dysfunction by history. CDU revealed penile vascular abnormalities in 76.5%. In Group I, veno-occlusive dysfunction (VOD) was observed in 40% while mixed (arterial + venous) vascular disease was diagnosed in 10%. Penile vascular disease was detected in 87.5% patients in Group II (ie VOD in 28%, arterial disease in 9.3% and mixed vascular disease in 50%). The prevalence of arterial disease on CDU among Peyronie' patients with erectile dysfunction (59.3%) was significantly (P < 0.05) higher then it was among potent patients (10%). The prevalence of pure VOD was almost similar for patients with or without erectile dysfunction (P < 0.05). We conclude that penile vascular abnormalities can be observed in 76.5% on cases with Peyronie's disease by CDU and this ratio significantly increases to 87.5% among cases with erectile dysfunction by history. Our data also indicate that arterial disease, accounts for much of the diminished rigidity in men with Peyronie's disease. PMID- 11424964 TI - Kallmann's syndrome: clues to clinical diagnosis. AB - Five cases of Kallmann's syndrome are presented, out-patients with microtestes, hypogonadotropic hypogonadism and complete anosmia. The final diagnosis was made only when they were aged between 17 and 26 (mean 21 years), although they had been seen by several physicians before: 3 for cryptorchidism and 3 for absence of spontaneous puberty; 2 had a positive family history, and 4 of the 5 patients or their parents admitted that they were aware of the fact that their sense of smell was completely absent, but they did not mention it spontaneously. PMID- 11424965 TI - Penile fractures: the successful outcome of immediate surgical intervention. AB - The data of 60 patients admitted to Mansoura Urology and Nephrology Center with penile fractures and treated by immediate surgical repair were reviewed with respect to their presentation, investigations, operative and post-operative details. Forty-nine patients were followed up regarding penile curvatures, plaques and erectile function. Patients reporting decreased erectile function were further assessed by evaluating their response to intracavernous injection of PGE1 and by penile color duplex Doppler ultrasonography. All of our patients had the classic clinical presentation of penile swelling and ecchymosis. Only five patients had accompanying urethral rupture. Penile ultrasonography was used to confirm the diagnosis in 23 patients. Immediate exploration was done using subcoronal circumferential incision in about two-thirds of the cases. All tunica albuginea ruptures were unilateral except one case which was bilateral. Interrupted absorbable sutures were used for repair in most of the patients. Urethral repair was done in five cases. Delayed complications were detected in only six cases (12.2%) in the form of mild penile curvature on erection, plaques and/or mild erectile dysfunction. Intracavernous injection (ICI) of PGE1 and penile duplex Doppler showed a normal pattern in three patients with erectile dysfunction while the fourth showed incompetent veno-occlusive mechanism. Psychosexual consultation was required for two of these patients while the third was successfully managed by self-ICI of PGE1. We conclude that the excellent outcome of our patients parallels other reports of early surgical repair regarding low morbidity, short hospital stay and rapid functional recovery. There is a low incidence and degree of erectile dysfunction among repaired patients; however, it should be thoroughly investigated and properly managed. Ultrasonography is easy and helpful; however, the more invasive cavernosography and/or magnetic resonance imaging are indicated when the case is atypical, or the diagnosis of rupture of tunica is suspicious. PMID- 11424966 TI - The anocavernosal erectile dysfunction syndrome. II Anal fissure and erectile dysfunction. AB - A previous study has demonstrated that the bulbocavernosus muscle (BCM) is a part of the external anal sphincter (EAS) [Shafik, Arch Androl, 1999]. It aids erection by compressing the penile bulb and the dorsal penile vein, and acts as a 'suction-ejection' pump in the ejaculatory process. Being a part of the EAS, the BCM is assumed to be involved in the different EAS pathologies. A recent study showed that erectile (ED) and ejaculatory dysfunction in 16 men with fecal incontinence (FI) after an anal fistula operation was cured after sphincteroplasty [Shafik, in press]. This article investigates the erectile and ejaculatory status in patients with anal fissure. The study comprised 32 men with acute anal fissure (mean age 36.7 +/- 8.2 s.d. years), 21 with chronic anal fissure (mean age 38.8 +/- 10.3 s.d. years), and 10 healthy volunteers (mean age 35.2 +/- 7.3 s.d. years). Erectile dysfunction occurred in all men with an acute fissure and in 16 of the chronic fissure patients; erection had been normal before fissure occurrence. The volunteers had normal erection. The anal pain radiated to the penis and was exaggerated on erection and penile thrusting. Erectile dysfunction investigations showed normal results. The electromyographic (EMG) activity of the external and internal (IAS) anal sphincters and the BCM as well as anal, penile bulb and cavernosal pressures were recorded. The acute fissures were treated conservatively and chronic ones by internal anal sphincterotomy. The patients were followed for mean periods of 17.3 +/- 3.6 s.d. months. The bulbocavernosus reflex as well as EMG activity of EAS and BCM were normal, while the resting EMG activity of the IAS was increased. The anal pressure in the acute and chronic anal fissure was increased (P < 0.01, P < 0.05, respectively), while the bulbar and cavernosal pressures showed no significant changes. Fissure treatment effected cure of the fissure and the ED in 30/32 of the acute and in 19/21 of the chronic cases. Erectile dysfunction persisted in the four patients in whom the fissures did not heal. In conclusion, a relationship is suggested to exist between anal fissure and ED. The ED occurred in the presence of anal fissure and was normalized with fissure cure. The BCM and anal pain seem to play a role in the etiology of ED associated with anal fissure. PMID- 11424967 TI - The role of intraoperative antibiotic irrigation and postoperative antibiotic therapy for contaminated implantable prosthesis: in a rat model in vivo. AB - The aim of this study was to determine the usefulness of intraoperative antibiotic irrigation solution and long-term effective antibiotic therapy for the infected prostheses. Forty-five male Wistar albino rats were divided into three equal groups and a small piece of silicone prosthesis contaminated with Staphylococcus epidermidis was implanted into the scrotum. In the first group, the silicone pieces were irrigated with an antibiotic solution intraoperatively and antibiotic therapy was applied for 20 days postoperatively. The second group underwent only antibiotic therapy. In the third group (control) neither intraoperative irrigation nor postoperative antibiotic therapy was applied. Postoperative clinical infection was determined as follow-up. All implants were extracted 20 days after the implantation and cultured to observe the bacterial growth. In the first group, in 13 rats the cultures were negative and in two rats, the cultures revealed positive bacterial growth. In the second group, in four rats the cultures were negative, in five rats the cultures were positive and six rats revealed infectious findings. In the third group, 13 rats revealed infectious findings, and in the remaining two rats the cultures were positive. The differences between three groups are statistically significant (P < 0.05). We conclude that intraoperative antibiotic irrigation and postoperative antibiotic therapy are highly beneficial in the infected prosthesis surgery. PMID- 11424968 TI - Long-term results with Nesbit's procedure as treatment of Peyronie's disease. AB - The objective was to assess sexual function at long-term follow-up after the Nesbit operation for Peyronie's disease. One hundred and fifty seven out of 213 patients treated between 1986 and 1998 using the Nesbit procedure were reassessed by means of IIEF-5 questionnaire together with two questions about residual deformity and treatment satisfaction. The results from this questionnaire together with the patient case records constitute the basis of this paper. After an average 72-month follow-up subjective patient determination of satisfaction indicated that 87.9% were satisfied with the results of surgery, 136 patients (86.7%) had good erectile function (IIEF-5 > 21). Shortening of the penis (from 1.5 to 3 cm) occurred in 22 patients (14%), but only in 2 (1.3%) was intercourse difficult because of excessive shortening. In conclusion, the Nesbit's operation results in the greatest amount of patient satisfaction about sexual function. When penile shortening occurs, it has not been a significant problem for patients who are properly counselled. PMID- 11424969 TI - The Sexual Health Inventory for Men (IIEF-5): reply to Cappelleri and Rosen. PMID- 11424970 TI - The role of immunotherapy in the treatment of HIV. PMID- 11424971 TI - Immune restoration and CD4+ T-cell function with antiretroviral therapies. AB - OBJECTIVES: To review the current understanding of the details and mechanisms of immune restoration that follows administration of suppressive antiretroviral therapies to persons with chronic HIV-1 infection. SUMMARY: A first-phase cellular increase often includes increases in multiple circulating lymphocyte populations and is largely attributable to rapid redistribution of cells from lymphoid tissue. A second slower phase is largely comprised of naive cell increases that may reflect cells newly produced in the thymus. Improvement in CD4+ cell function is demonstrable but functional restoration is incomplete. Immunization can enhance the restoration of CD4+ cell-dependent responses, and the magnitude of restoration is related in part to the degree to which HIV-1 replication and immune activation are controlled. Despite the incomplete nature of immune restoration seen after suppression of HIV-1 replication in chronic infection, clinical benefits of these responses are reflected in decreased HIV-1 related opportunistic infections and mortality. The effects of these therapies on the occurrence of non-Hodgkins lymphoma are less apparent. CONCLUSIONS: Suppression of HIV-1 replication results in both laboratory and clinical evidence of immune restoration. Although incomplete, this immune restoration provides 'breathing room' to develop better-tolerated antiviral therapies and therapies designed to enhance immune function. PMID- 11424972 TI - Cellular immune response to human immunodeficiency virus. AB - Despite recent success with the use of highly active antiretroviral therapy, eradication of HIV type 1 appears beyond the capabilities of presently available therapy. Therefore, greater emphasis has been given to finding mechanisms that promote immunologic control of viral replication rather than eradication. Although the correlates of immune protection in HIV-1 infection remain undefined, increasing evidence indicates that HIV-1-specific cellular immune responses may play a critical role in antiviral control. Vigorous HIV-1-specific CD4+ T-helper cell and CD8+ cytotoxic T-lymphocyte responses may play a critical role in control of viral replication in the absence of antiretroviral therapy, which has been demonstrated in individuals with long-term nonprogressive infection. However, in chronic, progressive HIV-1 infection, virus-specific T-helper cell responses are typically weak or absent in all stages of disease, and HIV-1 specific cytotoxic T-lymphocyte responses wane over time, presumably due to the lack of HIV-1-specific T helper cells. Effective treatment of individuals during acute HIV-1 seroconversion syndrome appears to restore HIV-1-specific T-helper cell responses, which are otherwise only observed in persons with long-term nonprogressive infection. This observation, along with anecdotal reports of individuals successfully controlling viral replication after treating acute HIV-1 infection, provides immunologic rationale for structured treatment interruption and other immunotherapeutic approaches designed to augment HIV-1-specific immune responses. PMID- 11424973 TI - The potential role of interleukin-2 in HIV. AB - Interleukin-2 (IL-2) is a secretory cytokine produced by activated T cells that stimulates T cells, B cells, and natural killer cells to proliferate and release cytokines. In addition, IL-2 slows apoptosis of HIV-infected cells. Clinical studies have demonstrated that exogenous human recombinant IL-2 can be safely administered concurrently with potent antiretroviral therapy to HIV-infected patients. It was further demonstrated that recombinant human IL-2 therapy produces sustained increases in CD4+ cell number and function in patients with both early and late HIV disease. Further evaluation of the clinical efficacy of IL-2 in HIV-infected patients is expected to provide important information on the utility of recombinant human IL-2 in HIV disease. PMID- 11424974 TI - Low-dose daily interleukin-2 immunotherapy: accelerating immune restoration and expanding HIV-specific T-cell immunity without toxicity. AB - There is now a great deal of interest in therapies focused on improving the function of the immune system in the treatment of individuals infected with the HIV. Although the antiviral drugs effectively suppress replication of the virus, they cannot cure the infection. Therefore, it now appears that both antivirals and immune system stimulants will be necessary to maximally suppress residual latent virus, thereby allowing the discontinuation of the antivirals without relapse of detectable plasma virus. Interleukin-2 (IL-2) the first cytokine to be discovered at the molecular level has been used as a therapeutic in HIV infection, because it is critical for a normal functioning immune response. IL-2 is essential for the survival and proliferative expansion of antigen-activated T cells and natural killer (NK) cells, and also for promoting their differentiated functions of cytokine secretion and cytolysis. However, as IL-2 stimulates both the innate and acquired immune responses, when used as a therapeutic it can lead to severe toxicity when given in high doses. This review focuses on low dose, daily IL-2 therapy, used to accelerate the recovery of the immune system when viral replication is suppressed maximally with antivirals. In addition, the principles of the use of IL-2 to activate HIV-specific immune reactivity are discussed. At least two signals are required to promote the proliferative expansion and function of antiviral effector lymphocytes, HIV antigens and IL-2. PMID- 11424975 TI - Perspectives on inducing efficient immune control of HIV-1 replication--a new goal for HIV therapeutics? AB - OBJECTIVES: A goal for long-term therapy of HIV infection is immune control of virus replication rather than the somewhat unrealistic aim of complete viral elimination. This paper will review the evidence that the control of viral infection can be achieved by an active CD8+ T-cell-mediated response. DESIGN: This review will draw on both experimental and clinical sources to discuss the potential mechanisms of the immune control. RESULTS: Data indicate that HIV infection can be effectively controlled by HIV-specific CD8+ T-cell-mediated responses. In infected individuals, the development of active cytotoxic T lymphocytes (CTLs, as measured by lytic activity) is associated with the control of viral replication. Within the simian immunodeficiency virus infection model in rhesus macaques, strong CTL responses are similarly associated with effective viral control. In addition, depletion by antibodies of CD8+ T cells within infected macaques results in rapid increases in viral load. However, in most HIV infected individuals, the CD8+ T-cells response is inefficient at low antigen dose, probably due to the lack of an effective H V-specific CD4+ T-cell response. If this CD4+ T-cell response is lost due to viral induced anergy, rather than clonal deletion, such responses may be generated by interruptions in antiretroviral treatment, and/or therapeutic immunization in chronically infected patients. A strong immune response stimulated at low-antigen dose early during viral rebound may be critical in preventing accumulation of toxic viral products that might inhibit effective CD4+ T-cell responses. CONCLUSION: Immune control of HIV infection is a realistic goal. Understanding both the basic immune mechanisms of in vivo viral replication and identifying practical therapeutic regimens to activate HIV CD4+ and CD8+ T-cell responses may allow the development of efficient immune control of HIV replication in chronically infected patients. PMID- 11424976 TI - The impact of highly active antiretroviral therapy on HIV-specific immune function. AB - Highly active antiretroviral therapy (HAART) can suppress HIV type 1 plasma viremia to undetectable levels for up to 3 years or more. When the therapy is discontinued, viral rebound occurs in a majority of patients, indicating that HAART is unable to completely eradicate the virus. Initial calculations of the half-lives of the infected cells (estimated to be 14-21 days) suggested that only 3 years continuous HAART therapy would be necessary to achieve complete eradication; however, several studies have determined that the half-lives of chronically infected cells are in the order of 6-44 months. New estimates indicate that it may take as long as 60 years to eradicate the virus. Thus, there has been movement toward combining HAART with various means of augmenting and/or reconstituting the host's immune system, especially HIV-1-specific immune responses. The long-term goal is to discontinue HAART and permit the reconstituted immune system to contain whatever small amounts of the virus remain. PMID- 11424977 TI - Samuel B. Guze, MD, 18 October 1923-19 July 2000. PMID- 11424978 TI - Genomewide survey of panic disorder. AB - We completed a genome scan of 23 multiplex families of panic disorder. Ninety family members had DSM-III-R panic disorder, and another 23 had recurrent, spontaneous panic attacks that did not satisfy these criteria. We typed 469 markers from the CHLC map (ver 8c7) with an average intermarker distance of 10.3 cM. Two-point lod scores were calculated with both a dominant and a recessive model, and maps of lod scores < -2.00, assuming genetic homogeneity, were constructed by using DSM-III-R panic disorder as the affected phenotype. Lod scores were < -2.00 over 94-95% of the genome. The greatest lod score was 2.23 (theta = 0.15) at the D7S2846 locus, located at 57.8 cM on chromosome 7 according to the Marshfield Clinic map. Flanking markers analyzed in a nonparametric, multipoint analysis using GENEHUNTER resulted in an NPL score of 2.97 at 63 cM on the Marshfield map. This region lies within 15 cM from the D7S435 locus, where Knowles et al. [1998] obtained a lod score of 1.71 (theta = 0.10) for panic disorder (now 2.45 with the addition of new families; James Knowles, personal communication). Thus, the maximum evidence of linkage from two genome scans of panic disorder lies within a small region of chromosome 7p. PMID- 11424979 TI - Impaired attention as an endophenotype for molecular genetic studies of schizophrenia. AB - Attentional abnormalities have long been known to characterize patients with schizophrenia. The data discussed in this report suggest that impaired attention (at least as measured by a specific task, the Continuous Performance Test, Identical Pairs (CPT-IP) version) may also be an endophenotype of particular promise for use in molecular genetic studies of schizophrenia. This conclusion is based on findings indicating that the deficits in verbal and spatial attentional processing tapped by the CPT-IP are heritable, developmentally stable, independent of clinical state, and predict future spectrum disorders in the at risk offspring of parents with schizophrenia. PMID- 11424980 TI - No evidence for involvement of the human inducible nitric oxide synthase (iNOS) gene in susceptibility to typical migraine. AB - Migraine is a debilitating disorder affecting approximately 12% of Caucasian populations. The disease has a large genetic component, although at present the type and number of genes involved is unclear. Candidate gene studies may be useful strategies for identifying genes involved in complex diseases such as migraine, especially if the gene being examined contributes only a minor effect to the overall phenotype. Nitric oxide (NO) is emerging as a key molecule affecting the pain associated with migraine. Since NO synthase (NOS) enzymes catalyze the synthesis of NO, the genes that code for these enzymes are good candidates for migraine molecular genetic analysis. This study investigated the role of a functionally relevant bi-allelic tetranucleotide polymorphism located in the promoter region of the human inducible nitric oxide synthase (iNOS) gene in migraine etiology. A large group of migraine affected individuals (n = 261) were genotyped and compared with an age- and sex-matched group of unaffected controls (n = 252). Results of a chi-squared analysis indicated that allele distributions for both migraine cases and controls were not significantly different (chi2 = 1.93, P = 0.16). These findings offer no evidence for an allelic association of the tested iNOS polymorphism with the common forms of the disease and therefore do not support a role for this gene in migraine pathogenesis. PMID- 11424981 TI - Nonreplication of association between mu-opioid-receptor gene (OPRM1) A118G polymorphism and substance dependence. AB - In the present investigation we hypothesized the A118G (Asn40Asp) polymorphism of the mu-opioid receptor gene (OPRM1) as a particular vulnerability factor for heroin and alcohol dependence. Therefore, we tested this hypothesis in two independent large samples by two different methods: a case-control sample (comprising n = 287 heroin and n = 221 alcohol study subjects versus n = 365 nondependent controls) and a family-controlled sample of 111 parent-offspring trios of heroin-dependent study subjects and 75 parent-offspring trios of alcohol dependent study subjects to avoid stratification artifacts. In both patient samples and by both methods we were unable to corroborate the hypothesis of OPRM1 A118G polymorphism as a particular risk factor for any kind of substance dependence including opioid addiction. In addition, there was no significant association between the endophenotype of the individuals under study (e.g., comorbidity, severity of illness) and a particular genotype of OPRM1. PMID- 11424982 TI - Linkage studies suggest a possible locus for developmental dyslexia on chromosome 1p. AB - Eight extended dyslexic families with at least four affected individuals were genotyped with twelve genetic markers spanning the Rh (rhesus factor) locus. Eleven of these markers were located on the short arm and the other was on the long arm of chromosome 1. Five theoretically derived phenotypes were used in the linkage analyses: 1) phonemic awareness; 2) phonological decoding; 3) rapid automatized naming; 4) single word reading; and 5) vocabulary. In addition, a lifetime diagnosis of dyslexia was used as a phenotype. Both parametric and non parametric genetic analyses were completed. The results supported the importance of a putative locus on 1p. In addition, two-locus analyses assuming the interaction between a 1p locus and a 6p locus, previously shown to be of interest for dyslexia, were conducted. As a result, the nonparametric linkage (NPL) scores for rapid automatized naming and phonological decoding were significantly increased. In particular, the NPL scores for rapid automatized naming exceeded 5.0 for certain markers. These results provide strong evidence for separate but jointly acting contributions of the 1p and 6p loci to the reading impairments associated with rapid naming and suggestive evidence for a similar mechanism involving phonological decoding. PMID- 11424983 TI - Population-based association analyses of the HOPA12bp polymorphism for schizophrenia and hypothyroidism. AB - HOPA is an Xq13 chromosome gene that codes for a RXR nuclear receptor co activator. In a prior study of the genetic basis of schizophrenia, we showed that exonic polymorphisms in HOPA were associated with increased risk of schizophrenia and hypothyroidism in a large cohort of probands from New York. In an attempt to replicate these findings, we examined this relationship in a cohort of 173 schizophrenic probands (128 males and 45 females providing 218 alleles) from Iowa. Consistent with the prior findings, we found an increased rate of the HOPA12bP exonic polymorphism in schizophrenic probands compared with random newborn controls (9 of 218 alleles vs. 33 of 2,049 alleles, P < 0.02). Furthermore, retrospective review of the medical records showed that two of the nine probands possessing the HOPA12bp allele in whom thyroid function was assessed were hypothyroid compared with 6 of 164 probands possessing the normal HOPAwild allele(s) (P < 0.06). We conclude that the HOPA12bp polymorphism shows a nominally significant association with schizophrenia and a nominal trend for association with hypothyroidism in our study and that further studies are required to define the features of this syndrome and the molecular mechanisms of disease pathogenesis. PMID- 11424984 TI - Quantitative neural indicators of liability to schizophrenia: implications for molecular genetic studies. AB - While schizophrenia is substantially heritable, the disorder's molecular genetic basis remains elusive. These efforts have been hindered by an inability to detect nonclinically-penetrant carriers of the predisposing genes and by uncertainties concerning the nature of the non-genetic influences and the extent of locus heterogeneity. The "endophenotype" approach is an alternative method for measuring phenotypic variation that may facilitate the identification of susceptibility genes in the context of complexly-inherited traits. Here we describe the application of this method to measures of brain structure and function in samples of schizophrenia patients and their non-ill first-degree relatives (siblings and co-twins). Our results suggest that there are likely to be multiple heritable dimensions of the central nervous system pathology in schizophrenia, each under the influence of a partially distinct set of genes, one of which involves disturbances in the structure and functioning of frontal lobe systems involved in working memory and another of which appears to render the brain more susceptible to damage to subcortical systems involved in long-term memory following oxygen deprivation in utero. Measures sensitive to quantitative variation in these dimensions of the central nervous system compromise should allow non-penetrant gene carriers to become informative for genetic linkage and facilitate detection of different genetic loci contributing to discrete aspects of disease liability. PMID- 11424985 TI - Linkage disequilibrium for schizophrenia at the chromosome 15q13-14 locus of the alpha7-nicotinic acetylcholine receptor subunit gene (CHRNA7). AB - The transmission/disequilibrium test was used for fine mapping of the linkage of schizophrenia to the chromosome 15q13-14 region, the site of a candidate gene, the alpha7 nicotinic acetylcholine receptor subunit gene (CHRNA7), in parent child triads from the NIMH Schizophrenia Genetics Initiative families. This candidate gene was identified from neurobiological studies of deficits in schizophrenics of the inhibitory gating of the P50 auditory evoked potential. The neurobiological deficit was also used as a phenotype for subsequent linkage analysis. In the present study, significant genotype-wise disequilibrium (P < 0.007) was found at D15S165, a polymorphic simple sequence marker physically located within 1 megabase of both CHRNA7 and a partially duplicated, expressed sequence that includes exons 5-10 of CHRNA7. Replication of this result was found in an additional set of families. The results support this region as a chromosomal location involved in the genetic transmission of schizophrenia. PMID- 11424986 TI - Early neurobehavioral deficits as phenotypic indicators of the schizophrenia genotype and predictors of later psychosis. AB - Schizophrenia is a genetically complex disorder that requires sharper delineation of its phenotypic boundaries. Considerable attention has been devoted in recent family and high-risk studies to the identification of both subclinical and other phenotypes, such as neurobehavioral deficits, that may be indicators of the genetic liability to schizophrenia. In high-risk studies, candidate liability indicators that are evident by young ages are also evaluated for their ability to predict future schizophrenia or spectrum disorders. We report on assessments of verbal short-term memory, gross motor skills, and global attention administered in midchildhood to offspring of schizophrenic, affectively ill, and normal parents as predictors of adult psychoses and as possible indicators of schizophrenia-susceptibility genes. PMID- 11424987 TI - The New York High-Risk Project: observations on the rating of early manifestations of schizophrenia. AB - The neurodevelopmental paradigm of schizophrenia (Sz) proposes a lifelong process of brain pathology as the cause of the disorder. Investigating the early phenotype of Sz, we rated videotapes of children at risk for Sz for early manifestations of thought disorder and Sz-related symptoms, using the Scale for the Assessment of Thought, Language, and Communication and parts of a diagnostic interview designed for psychiatric assessment of children. We found that thought disorder as well as negative symptoms were elevated in children with adulthood Sz related psychosis. This discussion focuses on methodological problems that arise in making clinical judgments on early and still subtle functional deviations. PMID- 11424988 TI - Symptom dimensions in obsessive-compulsive disorder: toward quantitative phenotypes. AB - The authors review evidence of symptom dimensions in obsessive-compulsive disorder (OCD) and their potential value as quantitative phenotypes in genetic studies of OCD and related conditions. Preliminary evidence supports the existence of four separate symptom dimensions. A small series of clinical and family genetic studies support the validity of one or more of these dimensions. However, critical data concerning the distribution of these dimensions in the general population are lacking, and the hereditability of these traits has yet to be established. PMID- 11424989 TI - Refining the ADHD phenotype using affected sibling pair families. AB - Attention deficit hyperactivity (ADHD) is a highly heritable behavioral disorder characterized by symptoms of inattention, hyperactivity, and/or impulsivity. Detection of susceptibility genes underlying ADHD may benefit from refinement of the ADHD phenotype into components that reflect more specific gene to behavior pathways and through the reduction of etiological heterogeneity. Using affected sibling pair (ASP) families, we are examining familial clustering of ADHD symptoms, neurocognitive task performance, and the occurrence of comorbid conditions to attempt to refine the phenotype and/or identify clinical features that may be used to stratify ADHD subjects to reduce etiological heterogeneity. PMID- 11424990 TI - The broad autism phenotype: a complementary strategy for molecular genetic studies of autism. AB - The genetic liability for autism appears to be expressed not only as the full syndrome of autism, but in milder, qualitatively similar characteristics that collectively have been referred to as constituting the broad autism phenotype. Identification of components of the broad autism phenotype that segregate independently in relatives of autistic individuals may provide an index of genes that, when present together, may interact to produce autism. Inclusion of information on the broad autism phenotype in relatives, in linkage studies of autism, may provide a potentially important, complementary approach for detecting the genes causing this condition. PMID- 11424991 TI - Quantifying the phenotype in autism spectrum disorders. AB - Twin and family studies suggest that familial transmission in autism extends to a spectrum of social and behavioral deficits that characterize individuals who have significant impairments within the autism spectrum, but do not meet formal criteria for autistic disorder. Standardized diagnostic instruments, including the Autism Diagnostic Interview-Revised (ADI-R) and the Autism Diagnostic Observation Schedule (ADOS-WPS Edition), offer the opportunity to quantify deficits across the autism spectrum, controlling effects of language and cognitive delay, in individuals with significant impairments. It is suggested that quantitative measures of social reciprocity and repetitive behaviors and interests, with separate quantification of expressive language level and nonverbal intelligence, most accurately reflect the range of behavioral phenotypes in autism spectrum disorders. PMID- 11424992 TI - Polydiagnostic approaches to measuring and classifying psychopathology. AB - Modern explicit diagnostic criteria have greatly improved the reliability of diagnosis in genetic and other types of psychiatric research. However, there are several sets of different diagnostic criteria that can be used, particularly in the case of studies focusing on the major psychoses. These different criteria do not always agree well with each other and there is a further difficulty that reliability does not necessarily ensure validity. A polydiagnostic approach is advocated whereby research data are collected in such a way as to allow the application of competing definitions of disorder. This is accomplished in the OPCRIT system by "decomposing" diagnostic criteria into their component items. These can then be reassembled using algorithms based on the original criteria or used to generate novel categories or dimensions. This is necessary if psychiatric diagnoses are ultimately going to be refined and validated against biological criteria. PMID- 11424993 TI - Phenotypes and genetic analysis of psychiatric and neuropsychiatric traits. AB - A workshop was held at Rockefeller University entitled "Phenotypes and Genetic Analysis of Complex Traits." The purpose of the workshop was to examine phenotype definition for complex traits, in particular, psychiatric and neuropsychiatric traits. An additional goal of the workshop was to examine statistical genetic approaches that specifically address the oligogenic nature of psychiatric traits. An overview of topics that were addressed and discussed at the workshop is presented in this article. PMID- 11424994 TI - Endophenotypes as quantitative risk factors for psychiatric disease: rationale and study design. AB - Genetic influences on psychiatric disorders are well established. However, localization of the genes responsible for these effects has proved extremely difficult. One emerging strategy that may circumvent some of these difficulties is the use of quantitative risk factors, or endophenotypes, which are correlated with disease and may be closer to underlying genetic liability and to gene action than are diagnostic phenotypes. Genetic studies of quantitative endophenotypes require different sampling and analysis strategies than studies of disease state. The rationale for using quantitative risk factors as indicators of disease liability and the optimal study design for localizing genes influencing such risk factors are discussed. PMID- 11424995 TI - Phenotype definitions: some hidden issues in psychiatry. AB - Three hidden problems in phenotype definition in psychiatric genetic studies are described. 1. Data on physical conditions in family members are not routinely collected in genetic studies of psychiatric disorders. These data may reveal cosegregation with psychiatric disorders which can be included in genetic analysis when defining the phenotype. Data on physical conditions can be obtained by a combination of medical checklists, medical history, and medical records. 2. Detailed clinical narrative summaries should be prepared on family members even if the narratives are not part of the diagnostic data included in centralized cell repositories for future distribution to qualified genetic investigators. Means for retaining the narratives for future use, while protecting patient confidentiality, should be sought. The narratives include detailed information on the context, severity, and sequence of the symptoms and can provide phenotype descriptions for reconsideration of diagnosis in the future. 3. The selection of early onset psychiatric disorders for genetic studies does not automatically mean that affected children should be included in genetic studies for some psychiatric disorders. Screening for genetic studies among child psychiatric patients and inclusion of children as affected in pedigrees should proceed with caution until more data are available from longitudinal studies on the continuity between childhood and adult psychiatric disorders. PMID- 11424996 TI - Good diagnoses require good diagnosticians: collecting and integrating the data. AB - The use of sets of diagnostic criteria and excellent structured clinical interviews will not result in data that is adequate for valid diagnoses or for dimensional measures if the rater collecting the data does not have adequate clinical skills. In the absence of biological measures or tests we are dependent on the rater's diagnostic and interviewing skills-both with subjects about themselves and with family informants asked to provide information about others. Best estimate diagnosticians (who usually have more experience and training) cannot compensate for inadequate or incorrect information and may even be misled by the information provided to them. In reviewing videotaped interviews sent by "trained" and "experienced" raters who were seeking "certification," approximately 40% of the raters were judged to have failed to conduct an adequate diagnostic evaluation. Principal Investigators should make every effort to assure that their diagnostic data is being collected by experienced and well-trained professional raters and that the raters' ongoing work is subjected to close supervision and review. PMID- 11424997 TI - "Targeting" schizophrenia in mice. AB - The greatest challenge of schizophrenia research remains the identification of the multiple, common, interacting, and moderately penetrant mutations that interfere with the highly complex function of human brain and result to this devastating disease. The inaccessibility of the human central nervous system to experimental manipulations and the paramount difficulties in identifying genes for schizophrenia has led researchers to generate mouse models for candidate genes using gene-targeting approaches. Although such mouse models have proven very useful in deciphering the causes of several diseases of the central nervous system (such as neurodegenerative diseases), their use in dissecting the biology of schizophrenia is still in its infancy. We argue that progress in this direction depends highly on progress in human genetic studies and requires careful and critical interpretation of the accumulating data. PMID- 11424998 TI - Oxytocin and vasopressin as candidate genes for psychiatric disorders: lessons from animal models. AB - Multiple approaches should be taken to investigate the genetic bases of psychiatric disorders, including the consideration of candidate genes. Studies in animal models suggest that the genes encoding oxytocin, vasopressin, and their respective receptors should be considered in a candidate gene approach for psychiatric disorders involving social deficits, such as autism or social phobias. These neuropeptide hormones may mediate the rewarding nature of social interactions and have been implicated in social attachment and social recognition in several animal models. Mutations in genes unrelated to oxytocin and vasopressin have been shown to have secondary effects on neuropeptide function and subsequent behavioral phenotypes. Genetic analysis of polymorphisms and expression analysis of candidate genes implicated in animal models may prove useful for determining the molecular mechanisms underlying psychiatric disorders, particularly in cases where other techniques proven difficult. PMID- 11424999 TI - Diagnosis as a covariate in sib-pair linkage analysis. AB - A novel technique to detect significant covariates in linkage analysis using a logistic regression approach is illustrated. An overall test of linkage is first performed to determine whether there is significant perturbation from the expected 50% sharing in sib pairs. Covariates may include variables defined on the sib pair (multiple levels of diagnosis), covariates defined on the parents (parental diagnosis or gender of the transmitting parent), or indicators of study when analyzing multiple datasets to examine heterogeneity. A detailed example using a hierarchical diagnosis of severe, intermediate, mild, and unaffected is provided. This permits testing whether sib allele sharing differs by level of diagnosis and allows the use of discordant pairs to protect against marker allele misspecification. PMID- 11425000 TI - Unbiased discordant sib-pair tests when parental genotypes are missing. AB - Population admixture and stratification are potential sources of confounding in a variety of statistical analyses of genetic data. Many approaches to adjusting for such confounding have been developed in the context of association analyses. These approaches may generally be viewed as relying on nonparametric null hypotheses that specify certain, sometimes incompletely observed, conditional distributions; the conditional distributions are used to normalize test statistics to have expectation zero under the null hypothesis. Here, with a very simple example, it is shown that there is potentially information that is free of confounding, but that is not recovered by such normalized statistics. An approach presented here in the context of the simple example might be extended to methods for recovering information in more complex settings. PMID- 11425001 TI - Neural networks and disease association studies. AB - After a brief introduction to neural networks, specific applications are proposed for the analysis of association between single nucleotide polymorphisms and putative disease genes in case-control studies. PMID- 11425002 TI - Model-free vs. model-based linkage analysis: a false dichotomy? AB - The aim of this article is to dispel some misunderstandings concerning the concepts of model-free or nonparametric linkage analysis (usually meaning affecteds-only analysis) and model-based or parametric analysis (usually meaning classical lod scores) and the implications of these concepts for the linkage analysis of complex genetic diseases. I review basic principles and then briefly indicate actual research findings. Readers may be surprised to learn that, technically, lod score analysis is also nonparametric. All this taken together indicates that classical lod score analysis, performed under a couple of different sets of genetic assumptions and corrected for multiple tests, is often actually superior to model-free analyses, even when the true genetic model is complex and unknown. PMID- 11425003 TI - Simulation studies of detection of a complex disease in a partially isolated population. AB - Simulation studies were undertaken with POPGEN, a new population simulation program, to explore strategies for detecting loci underlying rare and common disorders in a small population that has been partially isolated for 10 generations. Haplotype-sharing analysis (HSA) and non-parametric linkage analysis (NPL) were applied to the simulated haplotype and pedigree data for 100 cases, 100 controls, and an average of 28 multiplex pedigrees from cases' families, for a 2-5 cM map of markers. When identity by descent (IBD) status was known (using unique founder marker allele designations assigned during simulation), a linkage disequilibrium (LD) signal could be detected under disease-generating models predicting relative risk to sibs of 11.8 (high-RR) or 2.67 (mod-RR). Detection was more difficult when marker alleles were down-coded to resemble microsatellites (heterozygosities 0.75-0.80). False-positive peaks on nondisease chromosomes were uncommon. NPL analysis was more powerful than HSA at this marker density using down-coded alleles and assuming availability of all affected relatives. LD mapping of common disorders is likely to require denser maps of highly polymorphic markers to approximate full IBD information. LD and linkage mapping provide independent information, and strategies that combine these two methods could be useful in studies of small isolated populations. PMID- 11425004 TI - Possible dosage effect of maternally expressed genes on visual recognition memory in Prader-Willi syndrome. AB - Seventeen patients with Prader-Willi syndrome (7 with paternal deletion of chromosome 15q11-q13 and 10 with maternal uniparental disomy [UPD]), and 9 controls performed a computerized visual recognition task. A series of color digital photographs were presented; most were presented twice, but the remainder appeared only once. Photographs presented twice were separated in their presentation by either 0, 10, 30, 50 or 100 intervening photographs. Subjects indicated whether each photograph had been presented previously. This procedure was implemented twice, once using photographs of foods, and once using photographs of nonfood objects. As the number of intervening photographs between the first and second presentation increased, subjects were less likely to remember having seen the photograph before. Performance by UPD subjects was less affected by increasing the number of intervening photographs relative to the other two groups, suggesting they had superior visual recognition memory. This raises the possibility of a beneficial effect of having two copies maternally expressed genes on chromosome 15. UBE3A is suggested as a possible candidate for this effect. PMID- 11425005 TI - Variation in the LRP-associated protein gene (LRPAP1) is associated with late onset Alzheimer disease. AB - The LRP-associated protein is involved in the amount of mature LRP expressed on liver and brain. LRP is the main ApoE receptor and also binds alpha2 macroglobulin (alpha2M), a protein that associates with the beta-amyloid protein (betaA). By binding to alpha2M, LRP is responsible for the clearance of secreted betaA, thus preventing fibril formation. Genetic variation at the APOE, A2M, and LRP genes has been associated with the risk of developing late-onset Alzheimer disease (LOAD). We genotyped 373 patients, 300 controls, and 100 healthy elderly controls for a common DNA-polymorphism at the LRPAP1 gene (Insertion/Deletion, intron 5). Homozygotes for the rare Insertion (I) allele were at a significantly lower frequency in patients compared with controls (P = 0.002; OR = 0.29; 95% CI = 0.13, 0.68), and in patients compared with healthy elderly controls (P = 0.0002; OR = 0.18; 95% CI = 0.07, 0.46). No patient with an age at the onset below 75 years was II (0 of 214) compared with 8 in the group above 75 years (8 of 159) (P = 0.0044), suggesting that this genotype delays the onset of the disease. According to our data, the variation at the LRPAP1 gene is associated with the risk of developing LOAD. This is in agreement with the role of the LRPAP1 protein in the amyloidogenic pathway. PMID- 11425006 TI - Familial aggregation of anxiety-related quantitative traits in generalized social phobia: clues to understanding "disorder" heritability? AB - Social phobia, particularly the generalized form, is strongly familial. Given the probable continuity from extremes of normative personality (e.g., shyness) to social phobia to personality disorder (e.g., avoidant personality disorder), it is unlikely that social phobia itself, at the level of an Axis I disorder, is transmitted. Rather, it seems more parsimonious, and in keeping with current notions about the structure and heritability of mental disorders, that one or more temperamental risk factors for social phobia is transmitted. The goal of this study was to explore this possibility by examining relevant quantitative traits in a family study of generalized social phobia (GSP). First-degree (n = 103) relatives of patients with DSM-IV GSP and 65 first-degree relatives of not socially phobic comparison subjects (NC) completed a panel of self-report questionnaires that included measures of trait anxiety, social anxiety, and personality. Regression analysis was used to examine associations between group membership (i.e., whether that family member was a first-degree relative of a GSP proband) and these measures. First-degree relatives of GSP probands scored significantly higher than first-degree relatives of not socially phobic probands on measures of trait anxiety and social anxiety and on the Harm Avoidance subscale of the TPQ. One large factor, accounting for 84% of the variance, was strongly associated with being a first-degree relative of a GSP proband. Quantitative traits elevated in probands with generalized social phobia are also elevated in their first-degree relatives. Future family and genetic studies of social phobia should consider the possibility that one or more traits (or some aggregation thereof) may better approximate the phenotype of interest. More extensive efforts at phenotype refinement should be undertaken before such studies proceed. PMID- 11425007 TI - Defining alternative phenotypes for genetic studies: what can we learn from studies of schizophrenia? PMID- 11425008 TI - 5'-untranslated region of the dopamine D4 receptor gene and attention-deficit hyperactivity disorder. AB - Recently the molecular genetic basis of attention-deficit hyperactivity disorder (ADHD) has been the focus of a number of studies with the majority of these investigating the role of dopamine system genes. A great deal of attention has been focused on the possible involvement of the dopamine D4 receptor gene (DRD4) following a report of an association of ADHD with the allele containing seven copies of the 48-bp repeat in the third exon. In this paper we extended the search for the molecular explanation for the observed association by testing three polymorphisms in the region 5' to the dopamine receptor D4 gene transcription start site for linkage to ADHD. We specifically targeted polymorphisms in the region 5' to the start site of transcription as DNA variants in this region could alter the transcription level of the gene and hence the phenotype. We did not observe significant evidence for biased transmission of any of the alleles at these three polymorphisms to ADHD probands using the transmission disequilibrium test. We conclude that these three polymorphisms are not related to the ADHD phenotype. PMID- 11425009 TI - Family-based association study of the serotonin transporter promoter region polymorphism (5-HTTLPR) in attention deficit hyperactivity disorder. AB - Low serotonin activity has been associated in both animal and human studies with measures of impulsivity, aggression, and disinhibited behaviors. Recently, a common 44-bp deletion in the promoter region of the serotonin transporter (5 HTTLPR) that results in reduced transcription and lower transporter protein levels was described. Toward unraveling a possible role of the 5-HTTLPR polymorphism in childhood disruptive behaviors, we examined this gene in attention deficit hyperactivity disorder (ADHD), a heterogeneous childhood disorder in which three phenotypes are recognized by DSM IV criteria: inattentive (type I), hyperactive-impulsive (type II), and combined type (type III). By using the haplotype relative risk design, a group of 98 triads (both parents and proband child) were tested for a possible association between 5-HTTLPR and ADHD. A significant decrease in the short/short 5-HTTLPR genotype was observed in the ADHD type III combined group (10.29% vs. 30.88%) compared with the HRR-derived control group (likelihood ratio = 9.62, P = 0.008, n = 68 triads). Similar results were observed when allele frequencies were compared (likelihood ratio = 3.81, P = 0.05, n = 136 alleles). These first findings should be interpreted cautiously until replicated in independently recruited clinical samples. PMID- 11425010 TI - Allelic variants of the alpha1a adrenoceptor and the promoter region of the alpha2a adrenoceptor and temperament factors. AB - Human personality traits are partially determined by genes. It has been suggested that the reward-dependence dimension assessed by the Tridimensional Personality Questionnaire (TPQ) is related to the central noradrenergic system. Our population-based association study tested the hypothesis that genetic variants of the adrenoceptor are associated with this personality trait. The alpha1a- and the alpha2a-adrenoceptor genotypes were determined for 198 healthy Han Chinese who had completed the TPQ. We found no significant differences for TPQ personality factor scores, including reward dependence and its subscales, for subjects showing different adrenoceptor genotypes. Our negative findings suggest that polymorphisms of the alpha1a adrenoceptor and of the promoter region of the alpha2a-adrenoceptor have no major effect on the reward-dependence personality trait as assessed by TPQ. PMID- 11425011 TI - An association study between polymorphism of L1CAM gene and schizophrenia in a Japanese sample. AB - L1CAM, a neural cell adhesion molecule, plays an important role in the development of the central nervous system. The human L1CAM gene is located in Xq28. Mutations in the gene are responsible for a wide spectrum of neurological abnormalities and mental retardation. Schizophrenia may result from early neurodevelopmental abnormalities. We screened 30 male and 30 female schizophrenic patients for their genomic sequence of the L1CAM gene in order to determine the DNA sequence variations. Three novel variations located in exon 18 (10564 G > A, GG/AA at codon 758), intron 11 (8575 A > C), and intron 25 (13504 C > T) were detected. An association study of the identified polymorphisms was then performed in a Japanese sample of 152 male and 115 female patients with schizophrenia and 121 male and 114 female control subjects. A statistically significant increase in the count of the 13504 T-allele was observed in the male patients, compared to the male controls, with no differences in the variations of exon 18 or intron 11. There was no statistically significant change in the distribution of allele or genotype of any variations in the female schizophrenics, in comparison with the female controls. These results suggest that the polymorphism in intron 25 plays a role in the genetic predisposition of male schizophrenia in the Japanese sample. PMID- 11425012 TI - Stochastic dynamic population model for northern corn rootworm (Coleoptera: Chrysomelidae). AB - A stochastic dynamic population model for the complete life cycle of northern corn rootworm, Diabrotica barberi Smith & Lawrence, is described. Adult population dynamics from emergence to oviposition are based on a published single season model for which temperature-dependent development and age-dependent advancement determine adult population dynamics and oviposition. Randomly generated daily temperatures make this model component stochastic. Stochastic hatch is 50+/-8%. A stochastic nonlinear density-dependent larval survival model is estimated using field data from artificial infestation experiments. A regional model of corn phenology is estimated to incorporate the effect of dispersal on adult mortality. Random daily weather is generated using parameters for Brookings, SD. Model performance is evaluated with deterministic simulations, which show that the population converges to zero unless adult mortality is reduced by the availability of corn pollen from the regional model of corn phenology. Stochastic model performance is evaluated with stochastic daily weather, egg hatch, and larval survival in various combinations. Sensitivity analysis is conducted to evaluate model responsiveness to each parameter. Model results are generally consistent with published data. PMID- 11425013 TI - Bombus impatiens (Hymenoptera: Apidae): an alternative to Apis mellifera (Hymenoptera: Apidae) for lowbush blueberry pollination. AB - The pollination effectiveness of the commercially reared bumble bee Bombus impatiens Cresson, was compared in field studies to the honey bee, Apis mellifera L., for lowbush blueberry, Vaccinium angustifolium Ait. A preliminary study indicated that B. impatiens had potential as an alternative pollinator. In a 3-yr study, percentage fruit set, percentage harvested berries, berry weight, and seeds per berry were compared in blueberry fields stocked at 7.5 A. mellifera hives per hectare to 5, 7.5, or 10 B. impatiens colonies per hectare. Percentage of harvested berries (yield) was significantly higher in fields stocked with B. impatiens at 10 colonies per hectare. No other parameters measuring pollinator effectiveness were significantly different at 5, 7.5, or 10 colonies per hectare. Flower handling time was significantly faster for B. impatiens and it more frequently collected blueberry pollen. All parameters of pollinator effectiveness were similar for B. impatiens, A. mellifera, and native wild bees in a follow-up study. Overall, B. impatiens was a suitable alternative to A. mellifera. PMID- 11425014 TI - Quantitative assessment of biotic mortality factors of the European corn borer (Lepidoptera: Crambidae) in field corn. AB - Five treatments were used to exclude naturally occurring predators and parasitoids, based on body size and flight ability, to assess their effect on Ostrinia nubilalis (Hubner) populations on corn plaits. Two initial O. nubilalis egg densities (one egg mass and three egg masses per plant) were assigned to each treatment. Egg predation was higher in uncaged treatments than in caged treatments. Flying insect predators, primarily Coleomegilla maculata DeGeer (Coleoptera: Coccinellidae), reduced egg densities by 50%. Thirty-five to 84% of O. nubilalis larvae were infected with Nosema pyrausta (Paillot) (Microspora: Nosematidae). The incidence of Beauveria bassiana (Balsamo) Vuillemin (Deuteromycotina: Hyphomycetes), ranged from 0 to 21%, whereas larval parasitism, mainly by Macrocentrus cingulum Reinhard (Hymenoptera: Braconidae) ranged from 0 to 31%. In contrast to previous studies, this 3-yr field study documents that egg predation and larval infections of O. nubilalis were significant and consistent biotic mortality factors. PMID- 11425015 TI - Hazards of imidacloprid seed coating to Bombus terrestris (Hymenoptera: Apidae) when applied to sunflower. AB - Seed coating treatments of sunflower by the systemic insecticide imidacloprid was suspected of affecting honey bees and bumblebees. The hypothesis raised was whether imidacloprid could migrate into nectar and pollen, then modify flower attractiveness, homing behavior, and colony development. Our greenhouse and field experiments with Bombus terrestris L. were aimed at the following: the behavior of workers foraging on treated and control plants blooming in a greenhouse, the homing rate of colonies placed for 9 d in a treated field compared with colonies in a control field, and the development of these 20 colonies under laboratory conditions when removed from the fields. In the greenhouse, workers visited blooming heads of treated and control plants at the same rate and the mean duration of their visits was similar. In field colonies, analysis of pollen in hairs and pellets of workers showed that in both fields 98% of nectar foragers visited exclusively sunflowers, whereas only 25% of pollen gatherers collected sunflower pollen. After 9 d, in the control and treated field, 23 and 33% of the marked foragers, respectively, did not return to hives. In both fields, workers significantly drifted from the center to the sides of colony rows. During the 26 d period under field and laboratory conditions, the population increase rate of the 20 colonies was 3.3 and 3.0 workers/d in hives of the control and treated field, respectively. This difference was not significant. New queens were produced in eight colonies in either field. The mean number of new queens per hive was 17 and 24 in the control and treated field, respectively. Their mating rate was the same. It was concluded that applying imidacloprid at the registered dose, as a seed coating of sunflowers cultivated in greenhouse or in field, did not significantly affect the foraging and homing behavior of B. terestris and its colony development. PMID- 11425016 TI - Oviposition site and food preference of the green June beetle (Coleoptera: Scarabaeidae). AB - Relative preferences of green June beetle, Cotinis nitida (L.), adults and grubs for different organic fertilizers were determined in field and laboratory choice experiments. Six organic fertilizer treatments (low rate of broiler litter, high rate of broiler litter, cow manure, hay, Milorganite, or no fertilizer [the control]) were applied to sandy-loam soil and exposed to adults in 2.7 by 3.7 by 2.4-m screen cages. More eggs and larvae were found in pots treated with broiler litter (43%), cow manure (23%), and hay (30%) than in pots treated with Milorganite (4%) or no fertilizer (0%). Orientation preferences of third-instar grubs were tested in Y-tube and satellite olfactometers. Of the five treatments (broiler litter, cow manure, hay, Milorganite, and a blank control), preference was greatest for broiler litter and cow manure, but all organic fertilizer treatments were generally preferred over the blank control. These experiments suggest that use of organic fertilizers may result in higher densities of green June beetle grubs both by attracting the ovipositing females, and by acting as a food attractant for the mobile larvae. PMID- 11425017 TI - Relationship between cereal leaf beetle (Coleoptera: Chrysomelidae) egg and fourth-instar populations and impact of fourth-instar defoliation of winter wheat yields in North Carolina and Virginia. AB - Cereal leaf beetle, Oulema melanopus (L.), has become a serious pest of small grains in the mid-Atlantic region of the United States. Existing thresholds for implementing control measures allowed too much leaf damage and consequent yield loss to occur before recommending treatment. Information on beetle biology and crop response to injury, both prerequisites for developing new management strategies, was lacking for this region. A 3-yr project was initiated to generate an area wide cereal leaf beetle biological and yield impact database for winter wheat, and to evaluate the injury and yield loss potential of different population densities. Over the study period, beetle populations were evaluated at 26 winter wheat field locations in Virginia and North Carolina. Eggs and larvae, classified to instar, were counted twice each week from February to June. Replicated insecticide versus noninsecticide treatments were conducted at each location where leaf defoliation and yield were documented. Results showed that the relationship between 50th percentile egg and fourth-instar population estimates were in strong agreement (y = 0.36x - 0.01; r2 = 0.79). Potentially detrimental larval infestations were forecast before appearance of foliage injury from egg populations present during the stem elongation to flag leaf emergence developmental stages. A significant positive linear relationship between total fourth instar per stem population estimates and percent flag leaf defoliation was detected (y = 20.29x + 1.34; r2 = 0.60). A weaker but still significant relationship between the total fourth-instar population estimates and percent yield loss was found (y = 11.74x + 6.51; r2 = 0.26), indicating that factors in addition to flag leaf injury, primarily by fourth instars, also contributed to reduced yields. PMID- 11425018 TI - Droplet size and spray volume effects on insecticide deposit and mortality of heliothine (Lepidoptera: Noctuidae) larvae in cotton. AB - Effects of droplet size and volumetric application rate of tractor-applied lambda cyhalothrin (Karate Z 2.08 SC) on larval tobacco budworm, Heliothis virescens (F.), mortality and insecticide deposition on and within the canopy of mature cotton were explored by use of a laboratory bioassay incorporating field-sprayed cotton and a field trial incorporating natural insect infestations. Insecticide was applied in all combinations of three distinct droplet sizes and three volumetric application rates. Mortality of third-instar tobacco budworm occurring in leaf-disk bioassays was highly correlated with insecticide deposits (microg/leaf disk) at upper- and midcanopy levels. At the upper canopy level, mortality was negatively correlated with volumetric application rate and was not significantly correlated with droplet size. Deposit was negatively correlated with volumetric application rate at the lower plant level and larval mortality decreased with increasing droplet size. Results from this study do not support the recommendations of high volumetric application rates; and although droplet size was less influential than volumetric application rate in deposit and insect mortality, the data indicate a significant trend toward increased midcanopy larval mortality with smaller droplets. PMID- 11425019 TI - Native Lygus spp. (Heteroptera: Miridae) damaging introduced Hibiscus cannabinus in Italy. AB - Kenaf, Hibiscus cannabinus L, an introduced tropical fiber crop, is attacked in central Italy by the native mirid bugs Lygus rugulipennis Poppius and L. pratensis (L.), thus establishing novel insect-plant associations. Feeding by Lygus bugs damages the apical meristem, with consequent development of secondary stems and leaf tattering. In a laboratory experiment, both species caused apical meristem damage on potted kenaf. In a field experiment with caged plots, the percentage of damaged plants was significantly higher in plots artificially infested with multiple releases of Lygus spp. compared with a single release and with the protected control without Lygus spp. In naturally infested control plots, the percentage of damaged plants was intermediate. Plant height decreased as a function of the number of damaged meristems. Mean plant height was significantly lower in the multiple-release treatment compared with the single release treatment and the protected control. Dry biomass was significantly lower in the multiple-release plots compared with the protected control, and intermediate in the single-release plots. In a field experiment with three kenaf cultivars naturally infested by Lygus spp., plant height and stem diameter were significantly lower in damaged plants compared with healthy plants. Dry weights of plants, stems, and basts were also lower, depending on the cultivar, whereas the bast/core ratio was not affected. Stem lesions developed due to feeding or incisions made by ovipositing females. Lesions and eggs were significantly more abundant on plants with damaged apical meristems compared with healthy plants. Damage mechanisms by Lygus spp. and consequences of new crop introductions are discussed. PMID- 11425020 TI - Potential for mating disruption of Sparganothis sulfureana (Lepidoptera: Tortricidae) in cranberries. AB - The feasibility of disrupting mating of Sparganothis fruitworm with a sprayable microencapsulated formulation of (E)-11-tetradecenyl acetate (E11-14:Ac), the major pheromone component, was evaluated in New Jersey during 1996 and 1997 seasons. In both years, application of encapsulated E11-14:Ac, at 25-187.5 g (AI)/ha, reduced the incidence of mating of virgin females placed in treated plots relative to those placed in control plots. Pheromone trap catches were lower in pheromone treated plots, indicating that fewer male moths were able to locate the traps in treated plots. Larval density and fruit damage were significantly lower in plots treated with 62.5,125, or 187.5 g (AI)/ha of pheromone than in the untreated control. Air and foliage samples were collected to determine the air titers and foliage residuals of E11-14:Ac throughout the adult flight during 1996 and 1997. E11-14:Ac levels in air and foliage samples, declined sharply one wk after the pheromone application. However, detectable levels of E11-14:Ac were present in both air and foliage samples throughout the 3 to 4-wk period after the pheromone application. Multiple applications of pheromone at lower rates may be more effective in maintaining pheromone levels than a single dose at higher rates. These results suggest that mating disruption is a promising strategy to manage Sparganothis fruitworm in cranberries. PMID- 11425021 TI - Application technique and irrigation method affect imidacloprid control of silverleaf whiteflies (Homoptera: Aleyrodidae) on poinsettias. AB - Subirrigation systems are increasingly used to water and fertilize greenhouse crops. They also appear to be well suited for the application of systemic pesticides. We conducted two studies to look at interactive effects ofimidacloprid application technique and irrigation method on plant uptake of imidacloprid and whitefly control. Drench applications of imidacloprid resulted in much higher concentrations in the leaves than applications to the bottom of pots after 14 d. However, imidacloprid efficacy in subirrigated plants was better if the imidacloprid was applied to the bottom of the pot than when an equal amount was applied as a drench. In drip-irrigated plants, imidacloprid efficacy was greater after a drench than after an application to the bottom of the pots. A second study showed that drench applications to drip-irrigated plants result in high imidacloprid concentrations in the bottom of the canopy, whereas bottom applications to subirrigated plants result in a more even distribution of imidacloprid throughout the plant. Surprisingly, the high leaf imidacloprid concentrations in the bottom layer of drip-irrigated plants did not result in improved whitefly control. Imidacloprid efficacy was better in subirrigated, bottom-treated plants than in drip-irrigated, drenched plants. Overall, results from these studies indicate that imidacloprid is very effective when applied to the bottom of subirrigated pots. PMID- 11425022 TI - Apple damage, pest phenology, and factors influencing the efficacy of tebufenozide for control of obliquebanded leafroller (Lepidoptera: Tortricidae). AB - Fruit damage by obliquebanded leafroller, Choristoneura rosaceana (Harris), was examined on four different apple cultivars during 1997-1999 in heavily infested orchards in New York State. Inital fruit damage occurred 354+/-26 degree-days base 6 degrees C (DD6) after the first moth catch from the overwintering generation and continued to increase until after the typical spray season (approximately 1,300 DD6). In addition to apple damage by late instars, fruit damage occurred by early instars of the summer and overwintering generations. The insect growth regulator tebufenozide was used as a model insecticide to determine how a slow-acting insecticide and its relative toxicity to early (neonate) and late (fourth and fifth) instars may influence the efficacy of sprays for the control of the obliquebanded leafroller. Because tebufenozide is a slow-acting compound, bioassays were conducted to determine what percentage of the total mortality to neonates occurs at each 24 h interval until 10 d. Based on a polynomial regression, half of the total mortality to larvae at the LC25, LC50, LL90 and LC99 occurred at 7.2, 5.0, 4.1, and 3.0 d, respectively. Late instars were three times more tolerant to tebufenozide than neonates. PMID- 11425023 TI - Method of insecticide delivery affects horizontal transfer of fipronil in the German cockroach (Dictyoptera: Blattellidae). AB - Horizontal transmission of insecticide occurs when foragers contact or ingest an insecticide, return to the aggregation or nest, and translocate the insecticide to the shelter and its vicinity. Relatively more sedentary members of the population then contact or eat the translocated insecticide and die. We evaluated three different methods of delivering fipronil to adult male German cockroaches, Blattella germanica (L.), for their potential to cause such secondary mortality in various developmental stages of the cockroach. Adult males topically treated with 5 ng of fipronil (approximately LD99) caused low mortality in untreated nymphs and no mortality in untreated adults within the same aggregation. Males exposed to residual fipronil on a glass surface translocated more insecticide, resulting in higher mortality of cockroaches they contacted, but only early instars were affected and no adult mortality was observed. Ingested fipronil bait, however, was most effectively translocated, and caused high mortality of untreated adults and nymphs. Ingestion of fipronil also caused greater secondary kill compared with a topical application of 25 ng, approximately the same amount recovered from the exterior of males that ingested 1 mg of 0.05% fipronil bait. Secondary mortality in the untreated population was significantly affected by the duration of contact between the treated and untreated cockroaches, the quantity and freshness of excretions from the treated insects, and the accessibility of the secretions to untreated cockroaches. The mechanisms that cause secondary kill may include ingestion of excreted fipronil residues, cannibalism of bait-fed cockroaches, as well as contact with fipronil-contaminated substrates. PMID- 11425024 TI - Attractiveness of insecticide baits for cockroach control (Dictyoptera: Blattellidae): laboratory and field studies. AB - Several insecticide bait formulations were evaluated for their attractiveness to cockroaches in olfactometer assays in the laboratory and in trapping experiments in the field. Included in the assays were bait stations, gels, pastes, and a powder that contained one of the following active ingredients: abamectin, boric acid, chlorpyrifos, or hydramethylnon. There were significant differences among the baits in their attractiveness to the German cockroach, Blattella germanica (L.). In trapping experiments, Avert powder (abamectin), Maxforce station and gel, and Siege gel (all hydramethylnon) were consistently attractive to B. germanica adults and nymphs. Laboratory olfactometer assays with adult males confirmed these results and showed that nymphs were as responsive as males whereas females were less responsive. Our bioassays also demonstrate that attractiveness of bait can be dramatically affected by the age of the bait. One week of aging significantly reduced the attractiveness of Avert powder in both laboratory and field assays. Aging, however did not diminish the attractiveness of Maxforce gel, indicating that the formulation may be critical for retention of attractiveness of baits. Baits that were most attractive to the German cockroach were also the most attractive to nymphs and adults of the brownbanded cockroach, Supella longipalpa (F.). PMID- 11425025 TI - Evaluation of three methods for detecting permethrin resistance in adult and nymphal Blattella germanica (Dictyoptera: Blattellidae). AB - The insecticide resistance status of the German cockroach, Blattella germanica (L.), against permethrin was evaluated with three different bioassay methods. The LT50 and KT50 in adults were measured by glass jar test, the LT50 and KT50 in nymphs were assessed by glass petri dish-test, and the LD50 in adults and nymphs was measured by topical application. The results indicate that in all three bioassays, cockroaches showed different degrees of resistance to permethrin. Resistance ratios of 6.7-, 12.8-, and 7.5-fold for KT50, LD50, and LT50, respectively, were obtained for adult cockroaches compared with those from a susceptible strain. Similarly, resistance ratio values of 17-fold for KT50, 8 fold for LD50, and 6.4-fold for LT50 were obtained for nymphs compared with those from a susceptible strain. Although both adults and nymphs exhibited a similar level of susceptibility to permethrin, adults showed a higher degree of susceptibility to topical application than to the other exposure method. The order of sensitivity (based on resistance ratio values) was topical application (LD50) > LT50 > KT50 in adults. For nymphs, the resistance ratio in the knockdown test was higher than in other tests (resistance ratio = 17-fold). Therefore, the order of test sensitivity based on comparisons of resistance ratio values was KT50 > topical application (LD50) > LT50 in nymphs. Topical application exhibited a higher sensitivity than the LT and KT methods in adults. Thus, the results suggest that topical application and knockdown testing methods are the most sensitive assays for measuring resistance of adult and nymphal German cockroaches, respectively, to permethrin. PMID- 11425026 TI - Source-sink dynamics between transgenic and non-transgenic habitats and their role in the evolution of resistance. AB - The interaction of population dynamics and movement among two habitat types (toxic transgenic fields and nontoxic refuge fields) on the evolution of insecticide resistance was examined in two different simulation models. The two models were developed to test the hypothesis that increasing habitat grain from fine-grained to coarse-grained, and the resultant increase in nonrandom mating, would increase the rate of local adaptation, here the evolution of resistance. The first model, a complex, stochastic spatially explicit model, altered habitat grain by varying adult dispersal rates between habitat patches. In contrast to the expectation that increasing patch isolation and increasing the coarseness of the habitats would increase the rate of resistance evolution, intermediate levels of dispersal actually delayed resistance by as much as fivefold over the range of dispersal levels observed. Source-sink dynamics related to ovipositional patterns and the related population dynamics appear to explain the results. A simple deterministic model was developed to abstract out the separate impacts of mating and ovipositional behaviors. This model showed qualitatively the same results, although under similar assumptions it predicted much longer delays in resistance evolution. In this model, nonrandom mating alone always increased the rate at which insects adapted to transgenic crops, but nonrandom mating in combination with nonrandom oviposition could significantly delay resistance evolution. Differences between the two models may be due to the population regulation incorporated in the spatially explicit model. The models clearly suggest that resistance management programs using untreated refuges should not over-emphasize random mating at the cost of making the habitat too fine-grained. PMID- 11425027 TI - Biochemical mechanisms of resistance in strains of Oryzaephilus surinamensis (Coleoptera: Silvanidae) resistant to malathion and chlorpyrifos-methyl. AB - The acetylcholinesterase, carboxylesterase, and cytochrome P450 monooxygenase activities of three strains of Oryzaephilus srinamensis (L.) were examined to better understand biochemical mechanisms of resistance. The three strains were VOS49 and VOSCM, selected for resistance to malathion and chlorpyrifos-methyl, respectively, and VOS48, a standard susceptible strain. Cross-resistance to malathion and chlorpyrifos-methyl was confirmed in VOS49 and VOSCM. Acetylcholinesterase activity was not correlated to resistance among these strains. VOS49 and VOSCM showed elevated levels of carboxylesterase activity based on p-nitrophenylacetate, alpha-naphthyl acetate, or beta-naphthyl acetate substrates. PAGE zymograms showed major differences in caboxylesterase isozyme banding among strains. VOSCM had one strongly staining isozyme band. A band having the same Rf-value was very faint in VOS48. The VOS49 carboxylesterase banding pattern was different from both VOSCM and VOS48. Cytochrome P450 monooxygenase activity was based on cytochrome P450 content, aldrin epoxidase activity, and oxidation of organophosphate insecticides, all elevated in resistant strains. The monooxygenase activity varied with insecticide substrate and resistant strain, suggesting specific cytochromes P450 may exist for different insecticides. The monooxygenase activity of the VOS49 strain was much higher with malathion than chlorpyrifos-methyl as substrates, whereas VOSCM monooxygenase activity was higher with malathion than chlorpyrifos-methyl as substrates. Results are discussed in the context of resistance mechanisms to organophosphate insecticides in O. surinamensis. PMID- 11425028 TI - Variation in general esterase activity within a population of Haematobia irritans (Diptera: Muscidae). AB - Control of the horn fly, Hematobia irritans (L.), is generally dependent on chemical insecticides. However, the biology and behavior of the horn fly favors rapid development of insecticide resistance. To prolong the effectiveness of the insecticide option, information is required regarding the mechanisms of insecticide resistance. Metabolic hydrolysis of insecticides by esterases is a detoxification mechanism in many insect species. Measurement of general esterase activity within populations of horn flies may provide a diagnostic tool for resistance management. In this study we evaluated the amount of variation in general esterase activity within female and male horn fly samples from a population that had not been exposed to insecticides for 8 yr. We found considerable variation in general esterase activity within samples of each sex, with females demonstrating the greater variation. The observed variation is thought to be the result of age-structure dynamics within the population. The amount of inherent variation makes it difficult to detect small mean differences between populations, thus limiting the utility of general esterase assays. Thus, effective diagnosis of esterase-mediated resistance mechanisms can only be achieved by the identification of specific detoxification esterases and the design of assays, either biochemical or molecular, for their detection and measurement. PMID- 11425029 TI - Resistance to Diaphania hyalinata (Lepidoptera: Crambidae) in Cucumis species. AB - Diaphania hyalinata (L.), the melonworm, causes economic damage in cucurbit species in the Caribbean region. To control larvae feeding on leaves, frequent pesticide applications are needed, but this favors outbreaks of other pests. We looked for plant resistance to D. hyalinata in Cucumis melo L. and two wild Cucumis, C. metuliferus Meyer and C. pustulatus Naudin. Fifteen genotypes were observed using a 1-81 damage scale during 8 wk in two field trials in Guadeloupe, French West Indies. Among the C. melo genotypes, we observed different levels of damage (scores of 32-74 at the maximum damage date) ranging from partially resistant to susceptible genotypes. Both wild species scored <20 at the maximum damage date, and were more resistant in the field tests than the tested genotypes of C. melo. Five C melo genotypes and one C. pustulatus genotype were tested using a detached leaf test. Under controlled conditions, newly hatched larvae were fed with leaves from plants that were 5-9 wk old. Duration and mortality of immature stages were highly correlated. Strong antibiosis (long development and high mortality) was expressed by PI 140471 and Concombre Chien (two C. melo genotypes). For both genotypes, mortality was at least 87.9 versus 34% for Vedrantais, a susceptible genotype. Because antibiosis and field resistance were not correlated, we hypothesize that some genotypes expressed antixenosis in the field trials. In addition, possibilities of breeding for C. melo resistance to D. hyalinata is discussed. PMID- 11425030 TI - Biological effects of African yam bean lectins on Clavigralla tomentosicollis (Hemiptera: Coreidae). AB - The biological effects of affinity purified seed lectin from African yam bean, Sphenostylis stenocarpa (Harms), were tested on Clavigralla tomentosicollis (Stal) in an artificial seed system. S. stenocarpa was extracted from two African yam bean accessions: Enugu 95-3 and Enugu 98-2. Lectins from both accessions were highly toxic to the insect at 1.0% dry weight. For nymphs feeding on Enugu 95-3 and Enugu 98-2, survival ranged from 16.0 to 24.0% and 4.0 to 16.0% at 1.0 and 2.0% dry weight dietary lectin levels, respectively. From 4.0 to 8.0% dry weight of both lectins, no nymph survived up to 6 d after infestation. At 1.0% dry weights of Enugu 95-3 and Enugu 98-2 lectins, there was a significant delay in total developmental time (5.35 and 5.18 d), reduced survival (24.0% and 16.0%), and reduced growth (0.15+/-0.02 and 0.14+/-0.01) and resistance (61.85+/-9.78 and 57.79+/-3.80) indices, respectively. S. stenocarpa from Enugu 98-2 was the most toxic, with an LD50 of 0.43%, compared with an LD50 of 0.65% for S. stenocarpa from Enugu 95-3. The results of this investigation indicate that there exists in C. tomentosicollis physiological systems vulnerable to African yam bean lectins. PMID- 11425031 TI - Effect of planting dates and Bacillus thuringiensis corn on the population dynamics of European corn borer (Lepidoptera: Crambidae). AB - Field studies were conducted to determine how field corn, Zea mays L., phenologies in combination with transgenic Bacillus thuringiensis Berliner (Bt) corn and non-Bt (near isogenic) corn could affect egg laying by female European corn borer, Ostrinia nubilalis (Hubner), and subsequent larval injury. Transgenic Bt (events 176 and Bt11) and non-Bt corn was planted at three different times to assess the use of early- and late- planted Bt corn as a means for egg recruitment to these targeted planting dates. Plant growth stages, egg densities, and stalk tunneling was recorded at four locations in southwestern, central, and northern Iowa for three summers (1996-1998). No significant differences in egg densities were observed between Bt and non-Bt corn during the first and second generation for all three years. Significant differences did occur among planting dates. Between 50 and 100% of the eggs were laid in the early planting during the first generation. In addition, between 40 and 65% of the eggs were laid in the late planting for the second generation. Correlations between egg density and larval tunneling were inconsistent from year to year. Additional inconsistencies stemming from yearly phenological differences among sequential plantings and variable O. nubilalis populations increases the difficulty in recommending planting date adjustments as a practical management tool for European corn borer and Bt corn. PMID- 11425032 TI - Oxidative responses of resistant and susceptible cereal leaves to symptomatic and nonsymptomatic cereal aphid (Hemiptera: Aphididae) feeding. AB - The impact of the leaf-chlorosis-eliciting Russian wheat aphid, Diuraphis noxia (Mordvilko), and the nonchlorosis-eliciting bird cherry-oat aphid, Rhopalosiphum padi (L.), feeding on D. noxia-susceptible and -resistant cereals was examined during the period (i.e., 3, 6, and 9 d after aphid infestation) that leaf chlorosis developed. After aphid number, leaf rolling and chlorosis ratings, and fresh leaf weight were recorded on each sampling date, total protein content, peroxidase, catalase, and polyphenol oxidase activities of each plant sample were determined spectrophotometrically. Although R. padi and D. noxia feeding caused significant increase of total protein content in comparison with the control cereal leaves, the difference in total protein content between R. padi and D. noxia-infested leaves was not significant. Although R. padi-feeding did not elicit any changes of peroxidase specific activity in any of the four cereals in comparison with the control leaves, D. noxia feeding elicited greater increases of peroxidase specific activity only on resistant 'Halt' wheat (Triticum aestivum L.) and susceptible 'Morex' barley (Hordeum vulgare L.), but not on susceptible 'Arapahoe' and resistant 'Border' oat (Avena sativa L.). D. noxia-feeding elicited a ninefold increase in peroxidase specific activity on Morex barley and a threefold on Halt wheat 9 d after the initial infestation in comparison with control leaves. Furthermore, D. noxia feeding did not elicit any differential changes of catalase and polyphenol oxidase activities in comparison with either R. padi feeding or control leaves. The findings suggest that D. noxia feeding probably results in oxidative stress in plants. Moderate increase of peroxidase activity (approximately threefold) in resistant Halt compared with susceptible Arapahoe wheat might have contributed to its resistance to D. noxia, whereas the ninefold peroxidase activity increase may have possibly contributed to barley's susceptibility. Different enzymatic responses in wheat, barley, and oat to D. noxia and R. padi feeding indicate the cereals have different mechanisms of aphid resistance. PMID- 11425033 TI - Impact of Bt cottons expressing one or two insecticidal proteins of Bacillus thuringiensis Berliner on growth and survival of noctuid (Lepidoptera) larvae. AB - A series of laboratory assays were performed to compare the relative impact of commercial and experimental cultivars of cotton, Gossypium hirsutum (L.), expressing zero, one, or two insecticidal proteins of Bacillus thuringiensis Berliner, on several lepidopteran pests. Assays in which larvae were fed fresh plant tissue indicated that dual-toxin B. thuringiensis (Bt) cultivars, expressing both Cry1Ac and Cry2Ab endotoxins of B. thuringiensis, were more toxic to bollworms, Helicoverpa zea (Boddie), fall armyworms, Spodoptera frugiperda (J. E. Smith), and beet armyworms, Spodoptera exigua (Hubner), than single-toxin cultivars expressing Cry1Ac. Assays in which lyophilized plant tissue was incorporated into artificial diet also indicated improved activity of the dual toxin Bt cultivar compared with single-toxin plants. Both bollworm and tobacco budworm, Heliothis virescens (F.), growth was reduced by Bt cotton, particularly the dual-toxin cultivar. Although assays with lyophilized tissues were done using largely sublethal doses, bollworm survival was reduced by the dual-toxin cultivar. It appears that this newly developed Bt cotton expressing two toxins will be more effective and have a wider range of activity on these lepidopteran pests. PMID- 11425034 TI - A comparison of pitfall traps with bait traps for studying leaf litter ant communities. AB - A comparison of pitfall traps with bait traps for sampling leaf litter ants was studied in oak-dominated mixed forests during 1995-1997. A total of 31,732 ants were collected from pitfall traps and 54,694 ants were collected from bait traps. They belonged to four subfamilies, 17 genera, and 32 species. Bait traps caught 29 species, whereas pitfall traps caught 31 species. Bait traps attracted one species not found in pitfall traps, but missed three of the species collected with pitfall traps. Collections from the two sampling methods showed differences in species richness, relative abundance, diversity, and species accumulation curves. Pitfall traps caught significantly more ant species per plot than did bait traps. The ant species diversity obtained from pitfall traps was higher than that from bait traps. Bait traps took a much longer time to complete an estimate of species richness than did pitfall traps. Little information was added to pitfall trapping results by the bait trapping method. The results suggested that the pitfall trapping method is superior to the bait trapping method for leaf litter ant studies. Species accumulation curves showed that sampling of 2,192+/ 532 ants from six plots by pitfall traps provided a good estimation of ant species richness under the conditions of this study. PMID- 11425035 TI - Sampling for the sugarcane borer (Lepidoptera: Crambidae) on sugarcane in Louisiana. AB - A 3-yr study was conducted in 0.6- to 2.0-ha sugarcane fields throughout south Louisiana under varying sugarcane borer, Diatraea saccharalis (F.), density levels to determine the spatial dispersion of infestations and to develop a sequential sampling plan. Infestations of D. saccharalis were randomly dispersed. Infestation levels (percentage of stalks infested) ranged from 0.6 to 33.3%. Frequency distributions of the number of infested stalks indicated that the Poisson distribution best fit the data Tests of other distributions (negative binomial [aggregated], binomial [uniform], geometric, and hypergeometric) resulted in poorer fits. The sequential sampling plan devised, with lower and upper D. saccharalis infestation limits of 2 and 5% and 5 and 10%, required maximum average sample numbers of 7.1 and 5.5 (20-stalk samples), respectively, to make terminating management decisions. It is our assessment that implementation of these plans would decrease sampling effort by 50-60% when compared with sampling programs currently in use for D. saccharalis management decisions in Louisiana. PMID- 11425036 TI - Susceptibility of last instar red flour beetles and confused flour beetles (Coleoptera: Tenebrionidae) to hydroprene. AB - Last instar larvae of the red flour beetle, Tribolium castaneum (Herbst), and the confused flour beetle, Tribolium confusum Jacquelin du Val, were either exposed for 8-144 h on concrete treated with 1.9 x 10(-3) mg(AI)/per cm2 hydroprene, or continually exposed on concrete treated with 9.8 x 10(-4) to 1.9 x 10(-3) mg[AI]/per cm2 hydroprene. In both tests, larvae were exposed and held at 27 or 32 degrees C and 40, 57, or 75% RH. When larvae were exposed with no food to hydroprene for different time intervals, then transferred to untreated concrete containing flour, consistent effects were produced only at 144 h. At this exposure interval, the percentage of beetles arrested in the larval stage after 3 4 wk was generally greater at 75% RH compared with 40 and 57% RH, but there were no differences between species or temperature. The percentages of dead adult red flour beetles and live adults with morphological deformities were also greatest at 75% RH, and defects were more prevalent in red flour beetles than in confused flour beetles. When larvae were continually exposed to different concentrations of hydroprene on concrete that contained flour, the percentage of arrested larvae, dead adults, and live adults of both species generally increased with concentration. There were more deleterious effects at 75% RH compared with either 40 or 57% RH, and effects were more pronounced in the red flour beetle compared with the confused flour beetle. In both experiments, temperature effects were variable and inconclusive. Results indicate that continual exposure of last instar red flour beetle and confused flour beetle to hydroprene can limit population development, but exposure intervals of >6 d may be required for maximum effectiveness. PMID- 11425037 TI - Effects of stable flies (Diptera: Muscidae) on weight gains of grazing yearling cattle. AB - Differences in weight gains caused by stable flies, Stomoxys calcitrans (L.), on grazing yearling steer/calves averaged 0.2 kg per steer in a 3-yr study on canyon range pastures in West Central Nebraska, Stable fly numbers averaged 0.85 per front leg on treated calves and 3.64 per front leg on control calves. In 2 of the 3 yr after the grazing trials were completed, the calves were placed in a feedlot and fed a finishing ration. Compensatory gain did not occur in the feedlot after the stable fly stress was removed. PMID- 11425038 TI - Novel molecular therapeutic approach to cardiovascular disease based on hepatocyte growth factor. AB - Gene therapy techniques are being developed as potential treatments for cardiovascular diseases. During the past decade, many gene transfer methods including viral transfer techniques have been developed, and some are being applied clinically in human gene therapy studies. Recently, we have developed a novel gene transfer method mediated by Hemagglutinating Virus of Japan (HVJ) liposome, with which we have already reported several cases of successful gene transfer in vivo. Since the virus is inactivated by ultraviolet light, there is little potential for biological hazard with this method as compared to other viral gene transfer approaches. We also developed a novel strategy of gene therapy for cardiovascular diseases utilizing hepatocyte growth factor (HGF) which is an endothelial cell specific growth factor and an angiogenic growth factor. Based on these facts, we hypothesized that HGF may prevent restenosis after angioplasty through re endothelialization and myocardial infarction through induction of angiogenesis. The present results provide evidence of the efficacy of supplemental therapy with HGF by gene transfer in cardiovascular diseases. These data suggest the efficacy of novel molecular therapeutic approaches as gene therapy for cardiovascular diseases such as restenosis and myocardial infarction. PMID- 11425039 TI - Granulocyte/macrophage colony-stimulating factor plays an essential role in oxidized low density lipoprotein-induced macrophage proliferation. AB - We and other groups have recently demonstrated that oxidized low density lipoprotein (Ox-LDL) induces proliferation of macrophages in vitro. Since previous immunohistochemical studies demonstrated that macrophages and macrophage derived foam cells proliferated in situ in atherosclerotic lesions, it seems reasonable to expect that the Ox-LDL-induced macrophage proliferation might be linked to the development of atherosclerotic lesions. Thus, clarification of the molecular cascades of Ox-LDL-induced macrophage proliferation is expected to enhance our knowledge of the pathogenesis of atherosclerosis. Recently, we demonstrated that the activation of PKC leads to release into the culture medium of granulocyte/macrophage colony-stimulating factor (GM-CSF) which plays an important role in Ox-LDL-induced macrophage proliferation. In this review article, we mainly show the role of GM-CSF in the Ox-LDL-induced macrophage proliferation. Moreover, based on our recent findings, we summarize the Ox-LDL induced signaling pathway for macrophage proliferation. PMID- 11425040 TI - Markedly induced expression of LR11 in atherosclerosis. AB - Receptors belonging to the low density lipoprotein receptor (LDLR) superfamily play important biological roles in addition to mediating the lipoprotein metabolism. The recent discovery of a novel mosaic LDLR family member by us (Yamazaki H, Bujo H, Kusunoki J, Seimiya K, Kanaki T, Morisaki N, Schneider WJ, and Saito Y J Biol Chem 271: 24761-24768, 1996) and others, which we termed LR11, offers the opportunity to gain new insights into receptor multifunctionality. The expression of a 250-kDa mosaic LDLR family member, which we termed LR11 due to the presence of 11 ligand binding repeats, is markedly induced during the process of atherogenesis in two animal models. The highest induction of LR11 occurs in the intimal smooth muscle cells (SMCs) of atheromatous lesions. In agreement with the correlation of LR11 induction during increased cell proliferation in vivo, cultured SMCs showed a marked increase in LR11 expression in the proliferative phase. Furthermore, such proliferation-dependent expression of LR11 could be observed in a cultured neuroblastoma cell line, which was established to be a suitable in vitro model for proliferation and differentiation. Possible involvement of LR11 in the cellular proliferation sheds new light on the recently proposed novel functions of the LDL receptor gene family in atherosclerosis. PMID- 11425041 TI - Cholesterol-fed and transgenic rabbit models for the study of atherosclerosis. AB - The rabbit has been extensively utilized as an ideal model of atherosclerosis because of its size, easy manipulation, and extraordinary response to dietary cholesterol. The availability of spontaneously hypercholesterolemic model, Watanabe heritable hyperlipidemic rabbits (WHHL) and St. Thomas rabbits, has also provided insights into understanding human familiar hypercholesterolemia and atherosclerosis. With the advent of genetically engineered rabbits, transgenic rabbits have become a novel means to explore a number of proteins that are associated with cardiovascular diseases including atherosclerosis. To date, transgenes for human apo(a), apoA-I, apoB, apoE2, apoE3, hepatic lipase, lecithin: cholesterol acyltransferase (LCAT), lipoprotein lipase, 15 lipoxygenase, as well as for rabbit apolipoprotein B mRNA-editing enzyme catalytic polypeptide 1 (APOBEC-1), have been expressed in rabbits. In addition, human apoA-I, LCAT and apo(a) have been introduced into WHHL rabbits which have deficient LDL receptor function. All of these transgenes have been found to have significant effects on plasma lipoprotein metabolism or/and atherosclerosis. These studies have revealed new insights into the mechanisms responsible for the development of atherosclerosis. In this article, we provide a brief review on the rabbit model for the study of atherosclerosis with emphasis on transgenic rabbit models developed during the past few years. PMID- 11425042 TI - Japanese sisters associated with pseudohomozygous familial hypercholesterolemia and sitosterolemia. AB - Pseudohomozygous familial hypercholesterolemia is a rare condition of unknown etiology. Sitosterolemia is a rare autosomal recessively inherited disorder that is characterized by premature coronary artery disease, cutaneous xanthomas, and increased plasma plant sterols and 5alpha-stanols. Only a few cases of both sitosterolemia and pseudohomozygous familial hypercholesterolemia have been reported. In this study, we report two sisters with both conditions. With a low cholesterol diet (< 250 mg/day), serum cholesterol concentration decreased rapidly to an almost normal level and cutaneous xanthomas gradually regressed and finally disappeared; however, plant sterol levels did not change during the period. Plant sterols should be measured in patients considered to have pseudohomozygous familial hypercholesterolemia. The two conditions in this family may have been the results of a single gene mutation. The findings also indicate that low cholesterol diet therapy is effective for the treatment of hypercholesterolemia but not of sitosterolemia in this family. PMID- 11425043 TI - The gene expression profile of human umbilical vein endothelial cells stimulated by tumor necrosis factor alpha using DNA microarray analysis. AB - Stimulation of vascular endothelial cells by tumor necrosis factor alpha (TNFalpha) plays a critical role in the pathogenesis of inflammation and vascular diseases. Changes in the gene expression profile in cultured human umbilical vein endothelial cells (HUVEC) treated with TNFalpha was analyzed with high-density oligonucleotide arrays comprised of 35,000 genes. TNFalpha stimulation profoundly induced genes involved in signal transduction, leukocyte adhesion and chemoattraction. ICAM-1 mRNA (fold change 111.9) was most profoundly induced followed by TNFalpha receptor-associated factor 1 (TRAF1) (95.5), Bcl3 (71.8), IL8 (65.4), fractalkaine (62.4), E-selectin (48.0), lymphotoxin beta (41.3) and VCAM-1 (31.7). In addition to these previously known genes, 18 poorly characterized or novel genes known as ESTs profoundly induced by TNFalpha. Initial sequencing analysis identified three of these the genes for squalene epoxydase, chromodomain helicase DNA binding protein 4, and CLP respectively. Further analysis of these genes will provide important information about TNFalpha signaling and function in vascular endothelial cells. PMID- 11425044 TI - A Japanese patient with lipoprotein lipase deficiency homozygous for the Gly188Glu mutation prevalent worldwide. AB - We studied the molecular basis of familial lipoprotein lipase (LPL) deficiency in a new Japanese kindred. The proband was a four-month-old infant with severe hyperchylomicronemia. In postheparin plasma, LPL activity was virtually absent, although LPL mass was detectable. Single strand conformational polymorphism (SSCP) analysis showed an abnormal band with exon 5 of the LPL gene that was amplified by PCR from the proband's genomic DNA. DNA sequence analysis of the amplified fragment demonstrated that the proband was homozygous for a G-to-A change at nucleotide position 818 resulting in the substitution of glutamic acid for glycine at codon 188. Although this is among the first Gly188Glu mutations identified in Japanese, the missense mutation has previously been reported as a prevalent cause of familial LPL deficiency worldwide and has been proposed to have a common origin. However, DNA haplotype analysis with either restriction fragment length polymorphism (RFLP) or microsatellite markers revealed that the DNA haplotype of the proband was not identical to the haplotype previously reported as common to the other patients with the Gly188Glu mutation. These results add the Gly188Glu mutation to the growing list of LPL gene mutations underlying familial LPL deficiency in Japanese and indicate that the origin of the Gly188Glu mutation is not necessarily common but would be multicentric at least in part. PMID- 11425045 TI - Serum leptin and lipids in patients with thyroid dysfunction. AB - This work was undertaken to examine the relationship between thyroid hormone and serum leptin concentration. This study included 368 Japanese female subjects (27 were affected with pretreatment hyperthyroidism, 68 with hyperthyroidism during treatment, 19 with pretreatment hypothyroidism, 57 with hypothyroidism during treatment and 197 euthyroid control subjects) and 60 control male subjects. In the control group, serum leptin levels in males were lower than those recorded in females (mean +/- SD; 4.6 +/- 4.1 vs 9.5 +/- 6.4 ng/ml, p < 0.001). The leptin values correlated well with body mass index (BMI) and body fat mass (BFM) in both control male and female subjects (p < 0.001 for each). The serum leptin levels in pretreatment female patients with hyperthyroidism were significantly lower than those in the pretreatment patients with primary hypothyroidism and control female subjects (6.4 +/- 3.0 vs 9.7 +/- 6.3, 9.5 +/- 6.4 ng/ml; p < 0.05, 0.02, respectively), but after adjusting for BMI and BFM, the difference was mainly due to the significantly different BMI and BFM. Furthermore, serum leptin did not change significantly during the treatment in hyper and hypothyroidism. There was no correlation between serum leptin and thyroid hormones or lipids levels in female patients with thyroid disorders. Adiposity and gender were the major determinants of leptin concentration, but thyroid hormones did not appear to play any relevant role in leptin synthesis and secretion in human. PMID- 11425046 TI - The effects of pantethine on fatty liver and fat distribution. AB - Although the prognosis of fatty liver depends on its causes, we feel from our clinical experience that fatty liver with hypertriglyceridemia has a good prognosis and responds well to treatment. In this study, 600 mg/day of pantethine was administered to 16 outpatients with fatty liver and hypertriglyceridemia for six months or longer to examine whether the drug improved fatty liver using abdominal plain computed tomography (CT). Nine of the 16-pantethine patients were no longer diagnosed as having fatty liver after the study period. An chi2 test indicated the significant disappearance of fatty liver. At the same time, the visceral fat calculated from the CT image passing the umbilical region was also significantly reduced. On the contrary, the subcutaneous fat area tended to increase, so the ratio of the visceral-to-subcutaneous fat area was reduced significantly. This indicates triglycerides may be pooled in the body as hepato visceral fat and subcutaneous fat, and that pantethine may transfer fat from the liver and viscera to the subcutaneous tissue. This suggests that visceral fat deposition and fatty liver occurring with hypertriglyceridemia may have a common basis, probably excessive matrixes, and that pantethine may simultaneously improve the two conditions. PMID- 11425047 TI - Transgenic rabbits expressing human apolipoprotein (a). AB - Elevated plasma levels of lipoprotein (a) [Lp(a)] constitutes an independent risk factor for coronary heart disease, stroke, and restenosis. Over the past years, our understanding of the genetics, metabolism and pathophysiology of Lp(a) have increased considerably. However, the precise mechanism(s) by which this atherogenic lipoprotein mediates the development of atherosclerosis remains unclear. This is partly due to the lack of appropriate animal models since apolipoprotein (a) [apo(a)], a distinct component of Lp(a) is found only in primates and humans. Development of transgenic mice expressing human apo(a) has provided an alternative means to investigate many aspects of Lp(a). However, human apo(a) in transgenic mice can not bind to murine apoB to form Lp(a) particles. In this aspect, we generated transgenic rabbits expressing human apo(a). In the plasma of transgenic rabbits, unlike the plasma of transgenic mice, about 80% of the apo(a) was associated with rabbit apo B and was contained in the fractions with density 1.02-1.10 g/ml, indicating the formation of Lp(a). Our study suggests that transgenic rabbits expressing human apo(a) exhibit efficient assembly of Lp(a) and can be used as an animal model for the study of human Lp(a). PMID- 11425048 TI - Presentation and outcome of incarcerated external hernias in adults. AB - BACKGROUND: Incarcerated external hernias are the second most common cause of small-intestinal obstructions. The purpose of this study was to examine the presentation and management of incarcerated external hernia. METHODS: The records of 385 consecutive patients undergoing emergency surgical operation for incarcerated external hernias in a large volume teaching hospital between August 1996 and October 1999 were analyzed. The patients' ages ranged from 15 to 100 years (mean 55.1). There were more men than women (250 and 135, respectively), and 165 (42.9%) patients were over 60 years of age. Inguinal and umbilical hernias were encountered most frequently, in 291 (75.5%) and 48 (12.5%) patients, respectively. The intestine was resected in 53 patients, 31 of whom were over 60 years of age (58.5%). Two hundred fifty-two (84.9%) patients presented 48 hours or more from the onset of symptoms. Significant concomitant diseases were noted in 52 men and 19 women. RESULTS: The overall complication rate amounted to 19.5%, major complications 15.1%. The most serious postoperative complications were pulmonary and cardiovascular. Adult respiratory distress syndrome developed in 10 patients, and congestive heart failure developed in 14 patients. Postoperative mortality was 2.9%. Nine (81.8%) of the dead patients were older than 60. Nine (81.9%) of the dead patients were admitted to hospital more than 24 hours after incarceration. Mortality was high in patients with serious coexisting diseases whereas morbidity was linked with the duration of symptoms prior to admission. CONCLUSIONS: Older age, severe coexisting diseases, and late hospitalization were the main causes of unfavorable outcomes of the management of incarcerated hernias. PMID- 11425049 TI - Marketing strategy: an essential component of business development for academic health centers. AB - BACKGROUND: Historically, academic health centers (AHCs) have detached themselves from commercialism and entrepreneurism, viewing these activities as being inconsistent with many of their core academic values. Word-of-mouth promotion was their primary, if not sole, marketing strategy. Less emphasis was placed on preparing, pricing, distributing, and promoting these services to targeted audiences. Understanding customers' needs was not a top priority. METHODS: The marketing strategies and tools currently being developed and utilized by AHCs were reviewed. RESULTS: In an effort to attract customers and win contracts, AHCs are aggressively marketing themselves by designing new services, promoting those services much more intensely, restructuring the entire distribution system that delivers those services, and crafting pricing strategies that build in flexibility. With growing frequency, these marketing tactics are part and parcel of a carefully crafted data-driven strategic plan designed to meet the business development goals of the institution. CONCLUSIONS: In order to carry out their missions, AHCs have recognized that they can no longer rest on their "ivory tower" laurels. They must learn how to market themselves in a market economy. PMID- 11425050 TI - Challenging abdominal wall defects. AB - BACKGROUND: We propose a simple algorithm for management of patients with challenging abdominal fascial defects. METHODS: The medical records of 64 patients with complicated abdominal wall defects representing a consecutive series by a single surgeon over a 4-year period were reviewed. Group I patients presented with massive fascial defects and closed wounds. They were reconstructed with autogenous tissue using either the separation of parts (SOP) procedure or free tensor fascia lata (TFL) grafts. Group 2 patients had fascial defects with open wounds. Wound closure was first accomplished with either STSG or primary skin closure over viscera. These patients, now "converted" into patients with closed wounds, were reconstructed months later as in group 1. RESULTS: Average defect size was 320 cm2. Wound closure was achieved in one procedure in all patients with open wounds. Time to discharge after this procedure averaged 9 days. The only morbidity of wound closure was skin graft donor site pain. Average time from temporary staged closure with skin grafts to definitive closure with autogenous tissue was 5 months. Repair of closed fascial defects with autogenous tissue was performed in 51 patients. Average time to discharge after autogenous tissue repair was 6.6 days. Recurrence of hernia was noted in 2 (3.9%) patients with an average follow-up of 24 months. CONCLUSIONS: Treatment of challenging abdominal wall defects can be accomplished simply and safely utilizing the above surgical algorithm. Open wounds are converted into closed wounds and fascial defects are repaired with autogenous tissue. This treatment plan has proved to be effective in a wide variety of situations. PMID- 11425051 TI - Sonography of acute right side colonic diverticulitis. AB - BACKGROUND: To describe the prevalence and sonographic findings and ultrasound diagnostic accuracy of the right side colonic diverticulitis in patients having right lower abdominal pain with indeterminate nature. METHODS: A total of 934 patients with acute right lower abdominal pain of clinically indeterminate nature were referred for ultrasound examination (US). US studies were performed with 3.5 to 7.0 (or occasionally 10) MHz transducers using graded compression method. Twenty-three patients were finally diagnosed to have an uncomplicated acute diverticulitis of the right colon. The gray-scale sonographic images were reviewed. A retrospective study was undertaken to evaluate diagnostic contribution of US. RESULTS: The prevalence of acute right side colonic diverticulitis was 2.5% in patients with clinically indeterminate acute right lower abdominal pain. Locations of the inflamed diverticula include cecum in 6 patients, proximal ascending colon in 15 patients, and distal ascending colon in only 2 patients. Sonography detected 21 inflamed diverticula with 1 false positive and 2 false negative results. The most typical sonographic feature of an inflamed diverticulum of right side colon was a rounded or oval-shaped hypoechoic or nearly anechoic structure (52%) protruding out from the segmentally thickened colonic wall. Some of them might contain strong echoes representing gas or feces (43%), or stone in the lumen (5%). Regional pericolic or peridiverticular fat thickening was noted in 57% of patients, and segmental colon wall thickening in 38%. US examination yielded a sensitivity of 91.3%, a specificity of 99.8%, an overall accuracy of 99.5%, a positive predictive value of 95.5%, and a negative predictive value of 99.7%. A positive sonogram made the likelihood of acute right side diverticulitis 456.5 times greater compared with the pretest clinical impression. US differentiated acute right side colonic diverticulitis from acute appendicitis with a 100% accuracy. CONCLUSIONS: Ultrasound can be extremely useful in diagnosing acute right side colonic diverticulitis. Careful ultrasound evaluation of the right colon and the cecum may facilitate a correct diagnosis and help differentiate from acute appendicitis, and steer the surgeon to a more effective management. PMID- 11425052 TI - A multicentre collaborative study on the use of cold scalpel and electrocautery for midline abdominal incision. AB - BACKGROUND: Although studies in animals demonstrated a better wound healing after abdominal incision with cold scalpel than with electrocautery, clinical experiences did not confirm these findings. The purpose of this study was to compare early and late wound complications between diathermy and scalpel in gynecologic oncologic patients undergoing midline abdominal incision. METHODS: Patients undergoing midline abdominal incision for uterine malignancies were divided into two groups according to the method used to perform the abdominal midline incision: cold scalpel and diathermy in coagulation mode. Early and late complications were compared. Logistic regressions were used for statistical analysis. RESULTS: Nine hundred sixty-four patients were included, of whom 531 were in the scalpel group and 433 in the electrocautery group. Both groups were similar with respect to demographic, operative, and postoperative characteristics. Univariate analysis revealed a higher incidence of severe wound complications in the scalpel group than in the electrocautery group (8 of 531 versus 1 of 433, P <0.05). After adjustment for confounding variables (eg, age, body mass index) no differences were found between groups. CONCLUSIONS: Scalpel and diathermy are similar in terms of early and late wound complications when used to perform midline abdominal incisions. Therefore the choice of which method to use remains only a matter of surgeon preference. PMID- 11425053 TI - The indication of local excision for T2 rectal carcinomas. AB - BACKGROUND: Several researchers reported that local excision with or without postoperative chemo-radiation therapy is an alternative approach for sphincter preservation in patients with locally invasive rectal carcinoma. However, indications and long-term results have not yet been determined. METHODS: Seventy two patients with T2 colorectal carcinomas underwent bowel resection with regional lymph node dissection. The associations between lymph node metastasis (LNM) and clinicopathologic factors were examined with special reference to the presence or absence of moderate to severe degree of focal dedifferentiation or mucinous component at the invasive margin (unfavorable histology). RESULTS: Multivariate logistic regression analysis revealed that both sex and unfavorable histology were significantly associated with LNM (P = 0.0102, 0.0226, respectively). However, the associations between LNM and lymphatic invasion or tumor location were not statistically significant (P = 0.0947, 0.1738). CONCLUSIONS: When locally resected T2 rectal carcinoma specimens have unfavorable histology at the invasive margin, additional bowel resection with lymph node dissection should be recommended. PMID- 11425054 TI - Metoclopramide versus ondansetron in prophylaxis of nausea and vomiting for laparoscopic cholecystectomy. AB - BACKGROUND: Postoperative nausea and vomiting are significant problems in laparoscopic surgery. This double-blind, randomized, prospective trial compares the prophylactic use of metoclopramide, ondansetron, and placebo for the treatment of postoperative nausea and vomiting in patients undergoing outpatient laparoscopic cholecystectomy. METHODS: Two hundred thirty-two patients aged 18 to 73 years were randomized into three groups. Patients received intravenously 10 mg of metoclopramide, 4 mg of ondansetron, or placebo in a double-blinded manner prior to surgery. RESULTS: The incidence of nausea was 32% for metoclopramide, 45% for ondansetron, and 44% for placebo in the postanesthesia care unit or day surgery, which was not statistically significant. The incidence of vomiting was 8% for metoclopramide, 4% for ondansetron, and 22% for placebo in the postanesthesia care unit or day surgery. These differences were statistically significant when comparing both drugs to placebo but not when comparing both drugs to each other. CONCLUSION: Prophylactic administration of metoclopramide or ondansetron significantly reduces the incidence of postoperative vomiting for laparoscopic cholecystectomy, but neither drug was found to be significantly more effective than the other. Metoclopramide is a more cost-effective treatment. PMID- 11425055 TI - Effect of a resident self-study and presentation program on performance on the thoracic surgery in-training examination. AB - BACKGROUND: In this study we examine whether conversion from a didactic lecture format to a resident self-study and presentation program can improve performance on the Thoracic Surgery In-Training Examination (TSITE). METHODS: During the first 5 years, educational conferences were didactic lectures delivered by the attending thoracic surgery staff (group 1, n = 9 residents). During the second 5 years, residents prepared and delivered reviews from major textbook sources (group 2, n = 9 residents). Scores on the American Board of Surgery In-Training Examination (ABSITE) as a chief resident in general surgery were analyzed using one-way analysis of variance to assess fund of knowledge and test-taking skills prior to thoracic surgery training for the two groups. Scores on the TSITE during the first and second years of thoracic surgery training were recorded for each resident and analyzed using a paired t test. The data are expressed as the mean +/- standard deviation. RESULTS: Eighteen thoracic surgery residents over a 10 year period were involved in the study. ABSITE scores as a chief resident in general surgery did not differ between the two groups. Residents in group 1 improved their percentile rank from the first to the second year by a mean of 11%+/-12%, whereas those in group 2 improved their scores by a mean of 31%+/-21% (P < 0.05). CONCLUSIONS: When compared with a didactic lecture format, a resident self study and presentation program improves performance on the Thoracic Surgery In-Training Examination. This improvement in performance typically manifests during the second year of thoracic surgery training. PMID- 11425056 TI - Saphenoperitoneal anastomosis for resistant ascites in patients with cirrhosis. AB - BACKGROUND: Restriction of salt intake and diuretics combined with repeated paracentesis has been the mainstay of managing longstanding ascites. Peritoneal venous shunts have been employed in refractory ascites but are not without complication. We evaluated an autologous reversed segment of proximal long saphenous vein anastomosed to the peritoneum in management of patients with resistant ascites. METHODS: Eleven patients (8 male, median age 48 years, range 37 to 68) with tense refractory ascites associated with cirrhosis of the liver and portal hypertension underwent saphenous vein-peritoneal anastomosis by rotating the proximal vein cephalad which was anastomosed to peritoneum in the posterior wall of the inguinal canal. Ten of 11 procedures were performed under general anesthetic. RESULTS: Thirty-day mortality was 1 patient. Morbidity included transient hepatic encephalopathy in 4 (36%), minor wound hemorrhage in 3 (27%), fluid leakage in 7 (64%), and wound infection in 7 (64%). Hospital stay (median) was 16 days (range 11 to 23). In the short term (median of 9 months) significant reduction in body weight and abdominal girth was seen in 9 (90%), 6 (60%) were not on diuretics while 3 (30%) continued to remain on reduced doses of diuretic. Furthermore, 7 (70%) did not require paracentesis. At 2-year follow-up, 5 (45%) patients had died and 3 were lost to follow-up. The remaining 3 were all in active employment, 1 was off diuretics, and 2 were on reduced doses. All 3 patients maintained reduced body weights and abdominal girths compared with preoperative values. CONCLUSIONS: Saphenous-peritoneal anastomosis appears a simple, safe, and effective method of achieving long-term control of refractory ascites. The use of a biological shunt is an added advantage over prosthetic shunts for drainage of ascitic fluid. PMID- 11425057 TI - A vertical stomach reconstruction after pylorus-preserving pancreaticoduodenectomy. AB - BACKGROUND: Pancreatic fistula (PF) and delayed gastric emptying (DGE) is a leading cause of morbidity after pylorus-preserving pancreaticoduodenectomy (PPPD), occurring in 20% to 40% of patients. METHODS: Between August 1994 and March 2000, 30 consecutive patients underwent our modified PPPD and were evaluated on their incidence of PF and DGE. The major modification of our technique was an antecolic reconstruction and setting the transverse colon between pancreaticogastrostomy and duodenojejunostomy RESULTS: Operative time and blood loss were, respectively, 5.2+/-0.93 hours and 730+/-330 mL. Hospital mortality was 0%. Postoperative morbidity was 23%. Delayed gastric emptying and pancreatic fistula were observed in 3 (10%) and 0 (0%) of 30 patients. Nasogastric suction was required for 7+/-2 days, and a solid diet could be tolerated on postoperative day 11+/-4. CONCLUSIONS: The results show that our reconstruction can minimize DGE. PMID- 11425058 TI - Anatomical major resection versus nonanatomical limited resection for liver metastases from colorectal carcinoma. AB - BACKGROUND: Although systematic anatomical hepatic resection has been reported to improve patient survival in hepatocellular carcinoma, principles of hepatectomy procedure have not been clearly demonstrated in secondary hepatic malignancy. The purpose of the present study was to determine whether selection of surgical procedures for liver resection is associated with the pattern of tumor recurrence or patient survival. METHODS: During the period of 1980 through 1999, 174 cases underwent liver resection for hepatic metastasis from colorectal cancer. Of these, 96 underwent systematic anatomical major hepatic resection (anatomical group) and 78 cases underwent nonanatomical limited resection (nonanatomical group). Subset analysis of 115 patients with unilobar single or double tumors was also conducted. RESULTS: The overall 5-year survival rate of 174 patients was 43.2%. Univariate analysis did not show a significant difference in patient survival according to surgical procedure (anatomical group versus nonanatomical group). Operative morbidity and mortality rates were slightly higher in anatomical group. From the subset analysis in unilobar single or double tumors, anatomical major hepatectomy was unnecessary in 80.4% of the cases if the tumors were resectable by nonanatomical limited resection. Ninety percent of the ipsilateral recurrence, which could have been avoided if the first operation was anatomical hemihepatectomy, could undergo second hepatectomy with 5-year survival rate of 58.3%. CONCLUSIONS: There was not a significant difference in patient survival according to surgical procedure. To minimize surgical stress and operative risk, nonanatomical limited liver resection should be a basic surgical procedure for colorectal metastases. PMID- 11425059 TI - Hepatic ischemia-reperfusion injury. AB - BACKGROUND: The morbidity associated with liver transplantation and major hepatic resections is partly a result of ischemia-reperfusion injury. DATA SOURCES: The entire world literature on the subject was searched via Medline. Keywords included reperfusion injury, transplantation, liver resection, nitric oxide, endothelin, cytokines, Kupffer cells, ischemic/ischaemic preconditioning, and nuclear factor-kappa B. CONCLUSIONS: An imbalance between endothelin and nitric oxide levels results in failure of the hepatic microcirculation at the onset of reperfusion. Activation of nuclear factor-kappa B in the liver promotes proinflammatory cytokine and adhesion molecule synthesis. These result in oxygen derived free radical production and neutrophil recruitment, further contributing to cellular injury. Various therapeutic modalities acting on the above mediators have been successfully used to attenuate reperfusion injury in animal models of hepatic transplantation and resection. Application of the knowledge gained from animal models of hepatic ischemia-reperfusion to the clinical setting will improve the outcome of hepatic surgery. PMID- 11425060 TI - Predictive value of interleukin-8 and granulocyte elastase in pulmonary complication after esophagectomy. AB - BACKGROUND: We investigated whether or not interleukin-8 (IL-8) and granulocyte elastase (GE) can be associated with pulmonary complication after esophagectomy (the most common cause of postoperative death). METHODS: We measured serial changes in the IL-8 concentration and GE activity in the plasma and bronchoalveolar lavage fluid (BALF) of 17 patients who had undergone esophagectomy, and examined the relationship between these mediators and postoperative pulmonary complication. RESULTS: Pulmonary complication occurred in 6 patients (35%, Pneum+ group). Plasma IL-8 increased at the end of the surgery then decreased, but there was no significant difference between the Pneum+ group and the group without pulmonary complication (11[65%], Pneum- group). IL-8 and GE in BALF were significantly higher in the Pneum+ group than in the Pneum- group on days 1 and 3 after the operation. There was a significant and positive correlation between IL-8 and GE in BALF. CONCLUSIONS: Our results indicate that IL-8 and GE in BALF may be useful for the prediction of postoperative pulmonary complication. PMID- 11425061 TI - Pancreatic head resection with second-portion duodenectomy for benign lesions, low-grade malignancies, and early stage carcinomas involving the pancreatic head region. AB - BACKGROUND: Preservation of arterial blood supply to the duodenum and common bile duct during duodenum-preserving total resection of the pancreatic head is a major problem. We describe here a new procedure comprising pancreatic head resection with second-portion duodenectomy to overcome it. METHODS: The procedure was performed in 18 patients with benign lesions, low-grade malignancies, or early stage carcinomas involving the pancreatic head and with carcinoma of the middle bile duct or the gallbladder. The technique preserves the third portion of the duodenum by conserving the anterior inferior pancreaticoduodenal artery. The second portion of the duodenum is divided, followed by division of the lower bile duct and pancreatic neck. After resection followed by duodenoduodenostomy, there is a choice of two procedures: type A, pancreaticoduodenostomy and choledochoduodenostomy; or type B, pancreaticojejunostomy and hepatodochojejunostomy. RESULTS: There were no operative or hospital deaths (type A, 6; type B, 12). Postoperative complications occurred in 2 patients, but the others had an uneventful postoperative course. The quality of life of all patients has been satisfactory up to 36 months postoperatively. CONCLUSION: This procedure is a reliable option as an organ-preserving procedure for benign lesions, low-grade malignancies, and early stage carcinomas involving the pancreatic head. PMID- 11425062 TI - Clinical course, treatment, and multivariate analysis of risk factors for pyogenic liver abscess. AB - BACKGROUND: Pyogenic liver abscess is a threatening condition. The purpose of this study was to audit the clinical behavior and to analyze the risk factors. METHODS: One hundred and thirty-three patients treated in five hospitals during the years 1985 to 1997 were studied. By univariate and multivariate analysis we tried to identify any risk factor associated with complicated clinical course and complicated-related clinical course, and with hospital mortality. RESULTS: Sixty three patients (47%) were subjected to a percutaneous drainage, 45 (34%) were treated by open surgical drainage, and the remaining 25 cases (19%) received antibiotic therapy alone. Prognostic variables for a complicated clinical course were the presence of shock, low hemoglobin level, elevated prothrombin time, and polymicrobial infection. Shock, distress, low hemoglobin level, increased creatinine, and positive blood culture were significant predictors of a complicated-related clinical course. Concerning mortality, a biliary origin, shock, multiple abscesses, low hemoglobin level, and high concentration of blood urea nitrogen were independent predictors. CONCLUSIONS: Treatment of pyogenic liver abscesses should be tailored to each patient, however, the majority of them can be successfully treated with antibiotics and percutaneous methods. Those with signs of organ failure or septicemia should preferably be managed in the intensive care unit. PMID- 11425063 TI - Traumatic renal artery thrombosis. PMID- 11425064 TI - The carcinoembryonic antigen and its prognostic impact on immunocytologically detected intraperitoneal colorectal cancer cells. AB - BACKGROUND: Carcinoembryonic antigen (CEA) has been suggested to promote colon cancer progression. In this study we analyzed the prognostic impact of CEA expression on intraperitoneally detected single colon cancer cells. METHODS: Peritoneal lavage samples of 135 colorectal cancer patients were immunocytologically analyzed, including a staining of cellular CEA; serum CEA levels were measured; and 5-year survival rates were calculated according to immunocytological findings and CEA expression. RESULTS: The worst survival rate of 20% was found in patients suffering from CEA-expressing intraperitoneal tumor cells (P = 0.0006). The prognostic impact of an intraperitoneal tumor cell finding significantly increased when serum CEA levels were elevated: only 23% survived 5 years in contrast to a 85% 5-year survival rate of patients who neither had signs of dissemination nor showed elevated serum CEA values (P = 0.0010). CONCLUSIONS: This study shows that the determination of CEA expression improves the prognostic impact of an intraperitoneal tumor cell finding. PMID- 11425065 TI - Surgical Internet at a glance: continuing medical education. PMID- 11425066 TI - Predictors of positive margins after local excision of ductal carcinoma in situ. AB - BACKGROUND: This study aimed to examine the association between clinicopathologic parameters and positive margins in women with ductal carcinoma in situ (DCIS) treated by breast-conserving surgery (BCS). METHODS: We retrospectively reviewed the clinical, radiologic, and pathologic data of 100 women who had undergone BCS for DCIS in our center. RESULTS: Sixty-seven percent of patients presented via breast screening and 55% of all cases were diagnosed preoperatively on fine needle aspiration cytology ([FNAC] ie, C5). Overall, 45% of patients had clear margins after initial local excision. Positive margins showed a nonsignificant trend of association with distribution of microcalcifications (MCC), nonconsultant operating surgeon, inconclusive preoperative FNAC, presence of necrosis, and low specimen weight. There was a highly significant association between low grade DCIS (P = 0.003) and incomplete excision. There was no significant association with age, associated invasive focus, morphology of MCC, or with mode of presentation. CONCLUSION: Positive margins after local excision of DCIS are significantly associated with low nuclear grade, and preoperative determination of nuclear grade by core biopsy may have surgical implications. PMID- 11425067 TI - The diagnostic accuracy of mammography in the evaluation of male breast disease. AB - BACKGROUND: The role of mammography in the evaluation of male patients presenting with breast disease is controversial. This controversy is a function of the lack of specific data concerning the diagnostic accuracy of mammography when used in this clinical setting. The purpose of this study was to define the diagnostic accuracy of mammography in the evaluation of male breast disease. METHODS: One hundred and four prebiopsy mammograms from 100 patients with tissue diagnoses were read blindly by two independent radiologists, and placed into one of five predetermined categories: definitely malignant, possibly malignant, gynecomastia, benign mass, and normal. Radiologic/pathologic correlation was performed and the sensitivity (Sn), specificity (Sp), positive (Ppv) and negative predictive value (Npv), and accuracy (Ac) for each of the mammographic diagnostic category determined. RESULTS: The pathologic diagnoses were 12 cancers, including 1 patient with bilateral breast cancer, 70 cases of gynecomastia, 16 benign masses, and 6 normals. The accuracy data for the mammographic diagnostic categories are as follows: malignant (combined definitely and possibly malignant), Sn 92%, Sp 90%, Ppv 55%, Npv 99%, Ac 90%; and overall benignity (combined gynecomastia, benign mass, and normal), Sn 90%, Sp 92%, Ppv 99%, Npv 55%, Ac 90%. Six cancers (50%) coexisted with gynecomastia. CONCLUSIONS: Mammography can accurately distinguish between malignant and benign male breast disease. Although not a replacement for clinical examination, its routine use could substantially reduce the need for biopsy in patients whose mammograms and clinical examination suggest benign disease. PMID- 11425068 TI - Depth- and strain-dependent mechanical and electromechanical properties of full thickness bovine articular cartilage in confined compression. AB - Compression tests have often been performed to assess the biomechanical properties of full-thickness articular cartilage. We tested whether the apparent homogeneous strain-dependent properties, deduced from such tests, reflect both strain- and depth-dependent material properties. Full-thickness bovine articular cartilage was tested by oscillatory confined compression superimposed on a static offset up to 45%. and the data fit to estimate modulus, permeability, and electrokinetic coefficient assuming homogeneity. Additional tests on partial thickness cartilage were then performed to assess depth- and strain-dependent properties in an inhomogeneous model, assuming three discrete layers (i = 1 starting from the articular surface, to i = 3 up to the subchondral bone). Estimates of the zero-strain equilibrium confined compression modulus (H(A0)), the zero-strain permeability (kp0) and deformation dependence constant (M), and the deformation-dependent electrokinetic coefficient (ke) differed among individual layers of cartilage and full-thickness cartilage. HiA0 increased from layer 1 to 3 (0.27 to 0.71 MPa), and bracketed the apparent homogeneous value (0.47 MPa). ki(p0) decreased from layer 1 to 3 (4.6 x 10(-15) to 0.50 x 10(-15) m2/Pa s) and was less than the homogeneous value (7.3 x 10(-15) m2/Pa s), while Mi increased from layer 1 to 3 (5.5 to 7.4) and became similar to the homogeneous value (8.4). The amplitude of ki(e) increased markedly with compressive strain, as did the homogeneous value: at low strain, it was lowest near the articular surface and increased to a peak in the middle-deep region. These results help to interpret the biomechanical assessment of full-thickness articular cartilage. PMID- 11425069 TI - A treadmill ergometer for three-dimensional ground reaction forces measurement during walking. AB - This report describes new treadmill ergometer designed to measure the vertical and horizontal ground reaction forces produced by the left and right legs during walking. It was validated by static and dynamic tests. Non-linearity was from 0.2% (left vertical force) to 1.4% (right antero-posterior force). The resonance frequency was from 219 (right vertical direction) to 58 Hz (left medio-lateral direction). A calibration "leg", an air jack in series with a strain gauge, was developed and used to produce force signals comparable to those obtained during human locomotion. The mean differences between the force measured by the calibration leg and treadmill ergometer at 5 km h(-1) were 3.7 N (0.7%) for the left side and 6.5 N (1.2%) for the right. Measurements obtained during human walking showed that the treadmill ergometer has considerable potential for analysing human gait. PMID- 11425070 TI - The effect of interfacial parameters on cup-bone relative micromotions. A finite element investigation. AB - Achieving stability is a prerequisite for allowing bone to grow into the porous surface of non-cemented acetabular cups. The purpose of this study is to estimate the effects of interfacial characteristics on relative cyclical micromotion between cup and bone during gait in the immediate postoperative phase. The technique used is finite element analysis. Six models with different interfacial characteristics are created in order to study the effects of fixation technique. These include representation of a 1 mm press-fit, 2 mm press-fits (with and without an initial polar gap) and exact-fit conditions (with and without additional screw fixation). Although direct validation of the model has not been performed, the calculated micromotions under a static load of 1112 N are compared with appropriate experimental data. Generally, the model tends to underestimate micromotion and this underestimate is significant in the case of relative surface normal micromotion in polar regions for models with low- and no-interference. The most likely cause of this significant underestimate is a failure of the model to accurately represent penetration of rough contacting surfaces under compression. Other types of micromotion, although low, are within standard deviations reported by Kwong et al. (1994 Journal of Arthroplasty 9, 163-170). Quasi-static joint contact and muscle forces, representative of the stance phase of gait are then applied and maximum micromotions are found to occur consistently prior to toe off: this being the point of maximum force. With regard to the press-fit simulations, good cup-bone contact in the superior region of the interface is required for stability and the greatest micromotions occur in the models with the larger interference and larger polar gaps. In contrast to the press-fit models, muscle activity in exact-fit models influences the calculations. Specifically, the early activity of m.semimembranosus modelled causes opening of the peripheral seal. Taken together it is found that polar gaps reduce the stability of the model and lack of pre-compresssion in the periphery allows this region of the interface to be opened up. PMID- 11425071 TI - Investigating Murray's law in the chick embryo. AB - According to the optimization principle known as Murray's law, the blood vessel geometry at a bifurcation satisfies the relation alpha = (D3(1) + D3(2))/D3(0) = 1, where D0, D1, and D2 are the diameters of the parent and two daughter vessels, respectively. Previous investigations have shown that mature blood vessels adhere to this law fairly closely. The purpose of this study was to test Murray's law in the developing extraembryonic blood vessels of 2-4 day-old chick embryos. Vessel diameters were measured manually using image analysis software. The measurements for the group of all vessels at all studied stages (n = 449) gave alpha = 1.01+/ 0.34 (mean +/- SD), and the value of alpha is similar at all stages. These results indicate that Murray's law holds in the chick embryo, even before medial smooth muscle becomes functional, suggesting that blood vessels follow the same basic morphogenetic rules throughout life. PMID- 11425072 TI - A novel force transducer for the measurement of grip force. AB - A new strain gauge transducer has been developed to measure functional grip forces. The gripping area is a cylinder of diameter 30 mm and length 150 mm and simulates the handle of a number of devices, allowing a range of activities to be studied. The device measures radial forces divided into six components and forces of up to 250 N per segment can be measured with an accuracy of +/- 1%. The device therefore gives information about the magnitude and distribution of force around the cylinder during gripping, and has been shown to be a valuable research tool in a study of four different types of grip, providing valuable input data for biomechanical models. PMID- 11425073 TI - Biomechanical evaluation of two halo pin designs, with, and without, intact periosteum. AB - The presence of periosteum has been hypothesized to adversely affect halo pin penetration and performance (Voor, 1992. Ph.D. Dissertation, Tulane University, New Orleans, LA). However, biomechanical testing of halo pins has historically been conducted on bone specimens with periosteum removed. This may have lead to an unrealistic measure of biomechanical pin performance. Our study compares the biomechanical performance of two halo pin designs on bovine bone specimens with, and without, intact periosteum. The two pin designs included in this study were the conventional pin (Bremer Medical) with conical tip, and a newly released trochar-style pin (DePuy AcroMed). Results showed the mean peak load before failure of the trochar-style pin (mean +/- 95% confidence interval: 656+/-29 N) to be significantly higher than the conventional pin (517+/-53 N) on bone with intact periosteum (p = 0.001). With the periosteum removed, the mean peak load of the trochar-style pin (655+/-99 N) remained statistically the same (p = 0.987), while the mean peak load of the conventional pin (634+/-65 N) increased significantly (p = 0.026). Variation of the data of the conventional pin significantly decreased from 32 to 19% on removal of periosteum (sigma = 165-103 N, respectively, p = 0.0967), while variation of the trochar-style remained statistically the same at 30-29% (sigma = 193-188 N, respectively, p = 0.954). These results show that the trochar-style pin may be biomechanically superior to the conventional pin for vertical forces experienced during immobilization. The performance of this new pin style may also not be significantly affected by overlying soft tissue. Use of this new pin style may, therefore, improve overall stability and fixation of the halo apparatus. PMID- 11425074 TI - First cervical vertebra (atlas) fracture mechanism studies using finite element method. AB - Injury mechanisms and stress distribution patterns are important in the clinical evaluation of spinal injuries. Recognition and interpretation of the failure patterns help to determine spinal instability and consequently the choice of treatment. Although, the biomechanics responses of the atlas have received much attention, it has not been investigated using theoretical modeling. Mathematical techniques such as finite element model will provide further understanding to the injury mechanisms of the atlas, which is important for the prevention, diagnosis, and treatment of spinal injuries. In the present study, a detailed three dimensional finite element model of the human atlas (C1) was constructed, with the geometrical data obtained using a three-dimensional digitizer. Anterior arch, superior/inferior articular processes, transverse processes, posterior arch and posterior tubercule were modeled using eight-noded brick elements. Using the material properties from literature, the 7808-finite element model was exercised under three simulated axial compressive mode of pressure loading and boundary conditions to investigate the sites of failure reported in vivo and in vitro. This report demonstrates high concentration of localized stress at the anterior and posterior archs of the atlas, which agrees well with those reported in the literature. Furthermore, under simulated hyperextension, our results agreed well with the experimental findings, which show that the groove of the posterior arch is subjected to enormous bending moment. The close agreement of the failure location provided confidence to perform further analysis and in vitro experiments. These results may be potentially used to supplement experimental research in understanding the clinical biomechanics of the atlas. PMID- 11425075 TI - Machine vision photogrammetry: a technique for measurement of microstructural strain in cortical bone. AB - Understanding local microstructural deformations and strains in cortical bone may lead to a better understanding of cortical bone damage development, fracture, and remodeling. Traditional experimental techniques for measuring deformation and strain do not allow characterization of these quantities at the microstructural level in cortical bone. This study describes a technique based on digital stereoimaging used to measure the microstructural strain fields in cortical bone. The technique allows the measurement of material surface displacements and strains by comparing images acquired from a specimen at two distinct stress states. The accuracy of the system is investigated by analyzing an undeformed image set; the test image is identical to the reference image but translated by a known pixel amount. An increase in the correlation sub-image train parameter results in an increase in displacement measurement accuracy from 0.049 to 0.012 pixels. Errors in strain calculated from the measured displacement field were between 39 and 564 microstrain depending upon the sub-image train size and applied image displacement. The presence of a microcrack in cortical bone results in local strain at the crack tip reaching 0.030 (30,000 microstrain) and 0.010 (10,000 microstrain) near osteocyte lacunae. It is expected that the use of this technique will allow a greater understanding of bone strength and fracture as well as bone mechanotransduction. PMID- 11425076 TI - The role of the Womersley number in pulsatile blood flow a theoretical study of the Casson model. AB - The purpose of this Note is to clarify the meaning of the Womersley number alpha in pulsatile blood flow in small vessels. In particular. we explain why the use of alpha as aperturbation parameter to obtain approximate solutions of the Casson model (frequently used in the literature) is not appropriate. Using the techniques of dimensional analysis and scaling, we show that alpha is the product of the Reynolds and Strouhal numbers. Since the latter is very small for physiological flows, the result is that alpha < 1 even at relatively high values of the Reynolds number (i.e., for non-negligible inertia) and we validate our perturbation theory results by comparison with a numerical integration of the full model. Although this analysis is based on the Casson model, our method has general validity and may be applied to other models which describe more accurately the rheological properties of blood. PMID- 11425077 TI - In vitro steady-flow analysis of systemic-to-pulmonary shunt haemodynamics. AB - A modified Blalock-Taussig shunt is a connection created between the systemic and pulmonary arterial circulations to improve pulmonary perfusion in children with congenital heart diseases. Survival of these patients is critically dependent on blood flow distribution between the pulmonary and systemic circulations which in turn depends upon the flow resistance of the shunt. Previously, we investigated the pressure-flow relationship in rigid shunts with a computational approach. to estimate the pulmonary blood flow rate on the basis of the in vivo measured pressure drop. The present study aims at evaluating, in vitro how the anastomotic distensibility and restrictions due to suture presence affect the shunt pressure flow relationship. Two actual Gore-Tex shunts (3 and 4 mm diameters) were sutured to compliant conduits by a surgeon and tested at different steady flow rates (0.25-11 min(-1)) and pulmonary pressures (3-34 mmHg). Corresponding computational models were also created to investigate the role of the anastomotic restrictions due to sutures. In vitro experiments showed that pulmonary artery pressure affects the pressure-flow relationship of the anastomoses. particularly at the distal site. However, this occurrence scarcely influences the total shunt pressure drop. Comparisons between in vitro and computational models without anastomotic restrictions show that the latter underestimates the in vitro pressure drops at any flow rate. The addition of the anastomotic restrictions (31 and 47% of the original area of 3 and 4 mm shunts, respectively) to the computational models reduces the gap, especially at high shunt flow rate and high pulmonary pressure. PMID- 11425078 TI - Improved accuracy and reliability of sweepback angle, pitch angle and hand velocity calculations in swimming. AB - The estimation of forces in swimming using the quasi-static approach (Schleihauf, In: J. Terauds, J.P. Clarys (Eds.), Swimming III, International Series on Sports Sciences. Vol. 8, University Park Press, Baltimore, 1979, 70-109) has been popular in recent years as propulsion is an important determinant of performance. The aim of this study was to establish the accuracy and reliability of current and newly proposed procedures for the reconstruction of hand velocity, sweepback angle and pitch angle from underwater three-dimensional video analysis. A full scale mechanical arm capable of simulating a controlled and highly repeatable underwater phase of the front-crawl stroke was filmed for a set of five trials. A seven-point model of the arm and hand was then digitised at 25 Hz. Hand velocity, sweepback angle and pitch angle were calculated using the procedures of Schleihauf (1979), Berger et al. J. Biomech. 28 (1995) 125-133 and a newly proposed procedure (Lauder). Statistical comparisons were made between procedures to establish their relative accuracy and reliability throughout the stroke. The mean absolute error in measurement of hand velocity between points on the hand was very small (+/- 0.04 and +/- 0.06 m s(-1) in the x and z directions, respectively). The mean errors in sweepback angle and pitch angle were, respectively, 9.3 degrees and 7.6 degrees (Berger), 10.1 degrees and 8.1 degrees (Schleihauf and 10.7 degrees and 7.0 degrees (Lauder). Agreement between procedures showed the standard error between Schleihauf and Lauder to be the least (Schleihauf and Lauder, 0.4 degrees; Berger and Schleihauf, 1.3 degrees; Berger and Lauder; 1.6 degrees). The use of four points in the reconstruction of the orientation of the hand (Schleihauf and Lauder procedures) was shown to be less sensitive to errors in the digitising procedure. The reconstruction procedure proposed in this study (Lauder). further reduced the sensitivity to digitising error in the reconstruction of sweepback and pitch angles in swimming. PMID- 11425079 TI - Predicting systolic and diastolic aortic blood pressure and stroke volume in the intact sheep. AB - We developed a mathematical model describing the interaction between the heart and the arterial system. The model was constructed and tested on basis of invasive hemodynamic data in six sheep. Data from a first group of three animals (49 cardiac cycles) were used to assess a template time-varying elastance curve for the left ventricle, while the baseline steady-state data of a second group of three animals were used to assess reference cardiac and arterial parameters in sheep. The model is fully characterized by nine parameters, which were converted into 6 dimensionless numbers using the Buckingham pi theorem. The model was then used to generate LV pressure and volume and aortic pressure and flow for 86 conditions obtained by varying parameters 50 to 200% of their reference value. Systolic (SBP) and diastolic (DBP) blood pressure and stroke volume (SV) were determined from these model-generated curves and multiple linear regression analysis yielded the following expressions: SBP = Pisovolumic [0.638 - 0.0773 Emax C + 0.0507 RC/T] (r2 = 0.89); DBP = Pisovolumic [0.438-0.0712 Emax C + 0.0655RC/T] (r2 = 0.88) and SV = LVEDV [1.265-1.040 LVEDV/(LVEDV - Vd) + 0.125 Emax C-0.0777RC/T] (r2 = 0.93) with Pisovolumic = Emax (LVEDV - Vd), Emax and Vd being the slope and intercept of the end-systolic pressure-volume relation, R and C the total peripheral resistance and compliance, LVEDV the left ventricular end diastolic volume, and T the cardiac cycle length. These expressions were validated using data from the second group of three animals obtained during vena cava occlusion at baseline and during administration of dobutamine (61 cycles). The correlation between measured and predicted values was 0.98, 0.97 and 0.92 for SBP, DBP and SV, respectively. Compared to the measured values, SBP and DBP were, on average, underestimated by 5 and 6mmHg, respectively, and SV overestimated by 1.4 ml. We conclude that the derived expressions for blood pressure and stroke volume remain valid in the intact sheep for various hemodynamic conditions, and, taking into account their dimensionless form, may hold in other species and in humans. PMID- 11425080 TI - Optimal patterns for suturing wounds of complex shapes to foster healing. AB - Stress is one of the many biological factors that plays an important role in wound healing. It is therefore essential to analyze stresses around the wound closure theoretically, especially when no invasive/noninvasive technique to measure stress directly is available. The objective of this paper is to determine the regions of high stresses and the optimal pattern of suturing wounds of complex shapes. It is hypothesized that the optimal pattern of suturing wounds is that pattern which will produce minimum principal stresses. The finite element method (FEM) employing the basic equations of elasticity theory for orthotropic materials is utilized to compute the principal stresses and displacements resulting from suturing fusiform, elliptical and triangular wounds in human abdominal skin. The optimal suturing pattern for the triangular wound is determined. The average stress indices for varying suturing density are also determined which can provide useful clinical information for the surgeon. Since regions of high stresses in surgical closures produce adverse affects on healing and scar production, this work of predicting areas of high stresses is useful in indicating regions of slow healing in wounds. PMID- 11425081 TI - Oscillating fluid flow regulates cytosolic calcium concentration in bovine articular chondrocytes. AB - Mechanical loading is a well-known regulator of cartilage metabolism. This suggests that a loading-induced physical signal regulates chondrocyte behavior. Previous studies have focused on the effects of steady fluid flow on chondrocytes. In contrast to steady flow, loading induced fluid flow occurs in an oscillatory pattern and includes a reversal of flow direction with each loading event. In this study we examined the hypothesis that oscillating fluid flow increases cytosolic Ca2+ concentration ([Ca2+]i) in bovine articular chondrocytes (BAC) in a frequency-dependent manner and that the presence of serum affects this response. The aims of our study were to examine (1) whether BAC respond to physiologic oscillating fluid flow in vitro and compare these results to steady fluid flow, (2) the effect of fetal bovine serum on fluid flow responsiveness of BAC and (3) whether the response of BAC to fluid flow is flow rate and/or frequency dependent. [Ca2+]i was quantified using the fluorescent dye fura-2. BAC were exposed to steady, 0.5, 1, or 5 Hz sinusoidal oscillating fluid flow at five different flow rates in a parallel plate flow chamber. Our findings demonstrate that BAC respond to oscillating fluid flow with an increase in [Ca2+]i (p > 0.05), and furthermore, chondrocyte responsiveness to fluid flow increases with peak flow rate (p < 0.0001) and decreases with increasing frequencies (p < 0.0001). Finally, the presence of serum in the media potentiated the responsiveness of BAC to fluid flow (p < 0.0001). Our results suggest an important role for mechanical load-induced oscillating fluid flow in chondrocyte mechanotransduction. PMID- 11425082 TI - Age and gender differences in peak lower extremity joint torques and ranges of motion used during single-step balance recovery from a forward fall. AB - Previous studies have found substantial age and gender group differences in the ability of healthy adults to regain balance with a single step after a forward fall. It was hypothesized that differences in lower extremity joint strengths and ranges of motion (ROM) may have contributed to these observed differences. Kinematic and forceplate data were therefore used with a rigid-link biomechanical model simulating stepped leg dynamics to examine the joint torques and ROM used by subjects during successful single-step balance recoveries after release from a forward lean. The peak ROM and torques used by subjects in the study were compared to published estimates or measured values of the available maxima. No significant age or gender group differences were found in the mean ROM used by the subjects for any given initial lean angle. As initial lean angle increased, larger knee ROM and significantly larger hip ROM were used in the successful recoveries. There were substantial gender differences and some age group differences in peak lower extremity joint torques used in successful recoveries. Both young and older females often used nearly maximal joint torques to recover balance. Subjects' maximum joint strengths in plantarflexion and hip flexion were not good predictors of single-step balance recovery ability, particularly among the female subjects. PMID- 11425083 TI - An analysis of the effects of depth-dependent aggregate modulus on articular cartilage stress-relaxation behavior in compression. AB - An accurate description of the mechanical environment around chondrocytes embedded within their dense extracellular matrix (ECM) is essential for the study of mechano-signal transduction mechanism(s) in explant experiments. New methods have been developed to determine the inhomogeneous strain distribution throughout the depth of the ECM during compression (Schinagl et al., 1996, Annals of Biomedical Engineering 24, 500-512; Schinagl et al 1997. Journal of Orthopaedics Research 15, 499-506) and the corresponding depth-dependent aggregate modulus distribution (Wang and Mow, 1998. Transactions of the Orthopaedics Research Society 23, 484; Chen and Sah, 1999. Transactions of the Orthopaedics Research Society 24, 635). These results provide the motivation for the current investigation to assess the influence of tissue inhomogeneity on the chondrocyte milieu in situ, e.g. stress, strain, fluid velocity and pressure fields within articular cartilage. To describe this inhomogeneity, we adopted the finite deformation biphasic constitutive law developed by Holmes and Mow (1990 Journal of Biomechanics 23, 1145-1156). Our calculations show that the mechanical environment inside an inhomogeneous tissue differs significantly from that inside a homogeneous tissue. Furthermore, our results indicate that the need to incorporate an inhomogeneous aggregate modulus. or an anisotropy, into the biphasic theory to describe articular cartilage depends largely on the motivation for the study. PMID- 11425084 TI - Simulation of acute subdural hematoma and diffuse axonal injury in coronal head impact. AB - Coronal head impacts were simulated in a physical model, based on the hypothesis that acute subdural hematoma (ASDH) is related to cerebral vertex displacement and diffuse axonal injury (DAI) to local Green-Lagrange strain. The geometry of the 2D model was based on anatomical measurements taken from the MRI scans of 10 adult males. Silicone gel modelled the cerebrum, paraffin the CSF and elastic membranes the trabeculae of the sulci. Pendulum impacts gave peak angular acceleration of 7800 rad s(-2) in models with and without sulci. The motion of the gel and Green-Lagrange strain were calculated from tracked coordinates of Patrick markers. Worst-case bridging vein strains are produced on the contrecoup side and are approximately doubled by adding sulci. Given that axons in the corpus callosum are highly oriented, Green-Lagrange strain was resolved in the fibre direction. It is found to be close to the minimum principal strain, indicating a degree of natural, teleological protection for the axons. The data support the use of delta0peak as a suitable descriptor for the risk of DAI but not for ASDH. PMID- 11425085 TI - Local hemodynamics affect monocytic cell adhesion to a three-dimensional flow model coated with E-selectin. AB - Monocyte adhesion to the endothelium depends on concentrations of receptors/ligands, local concentrations of chemoattractants, monocyte transport to the endothelial surface and hemodynamic forces. Monocyte adhesion to the inert surface of a three-dimensional perfusion model was shown to correlate inversely with wall shear stress, but was also affected by flow patterns which influenced the near-wall cell availability. We hypothesized that (a) under the same flow conditions, insolubilized E-selectin on the model's surface may mediate adhesive interactions at higher wall shear stresses, compared to an uncoated model, and (b) pulsatile flow may modify the adhesion profile obtained under steady flow. An axisymmetric flow model with a stenosis and a sudden expansion produced a range of wall shear stresses and a separated flow with recirculation and reattachment. Pre-activated U937 cells were perfused through the model under either steady (Re = 100, 140) or pulsatile (Remean = 107) flow. The velocity field was characterized through computational fluid dynamics and validated by inert particle tracking. Surface E-selectin greatly increased cell adhesion in all regions at Re = 100 and 140, compared to an uncoated model under the same flow conditions. In regions where the cells near the wall were abundant (taper and stenosis), adhesion to E-selectin correlated with the reciprocal of local wall shear stress when flow was steady. Pulsatile flow distributed the adherent cells more evenly throughout the coated model. Hence, characterizing both the local hemodynamics and the biological activity on the vessel wall is important in leukocyte adhesion. PMID- 11425086 TI - Post-traumatic intrasplenic pseudoaneurysms with delayed rupture: color Doppler sonographic and CT findings. AB - Post-traumatic intrasplenic pseudoaneurysms are very rare in children. Since pseudoaneurysms may expand a splenic hematoma and cause delayed splenic rupture, early diagnosis and treatment are crucial. In this report, we describe the case of a 12-year-old boy with a delayed splenic rupture caused by a splenic hematoma containing 2 pseudoaneurysms. Abdominal sonography showed free intraperitoneal fluid and a mildly enlarged spleen with a large heterogeneous area occupying the upper half of the organ. Two anechoic lesions (15 and 4 mm) were seen inside the hematoma near the splenic hilum. Color Doppler sonography demonstrated turbulent arterial flow within the lesions, suggesting pseudoaneurysms. On CT, the lesions enhanced simultaneously with the splenic artery in the arterial phase of contrast enhancement. CT also showed an intrasplenic arterial branch leading to the larger of the 2 pseudoaneurysms. PMID- 11425087 TI - Reversal of diastolic plateau in partial testicular torsion. AB - Testicular torsion is a urologic emergency. The ability of color and power Doppler sonography to diagnose partial testicular torsion definitively remains uncertain. We present a case in which color Doppler findings were indeterminate and the diagnosis of partial testicular torsion was made on the basis of spectral waveform analysis. The characteristic findings in the spectral waveform were asymmetry between the 2 testes, with a higher intratesticular resistance index on the affected side, and reversal of the diastolic plateau on the affected side. PMID- 11425088 TI - Sonographic findings in brucellar hepatic abscess. AB - Brucellar hepatic abscesses are rare; to our knowledge, only 41 cases have been described in the literature. We report the case of a 29-year-old Spanish woman who was hospitalized because of fever, weakness, and weight loss. Abdominal sonography showed a hypoechoic lesion with central calcification. Brucella spp. were not isolated from an aspirate of the liver or blood cultures. The diagnosis was based on the association of characteristic sonographic features (central calcification and peripheral necrotic areas) and positive Brucella agglutination tests. The patient improved rapidly with antibiotic treatment. PMID- 11425089 TI - Prenatal sonographic diagnosis of axillary cystic lymphangiomas. AB - Cystic lymphangiomas are congenital malformations of the lymphatic system that occur most commonly in the nuchal region. Nuchal lymphangiomas are often referred to as cystic hygromas. Few cases of axillary cystic lymphangioma have been reported, but the prognosis of this lesion appears to be less grave than that of its nuchal counterpart. We present 2 cases of axillary cystic lymphangiomas that were diagnosed on prenatal sonography. In 1 case, prenatal sonography showed a multilocular, large cystic mass on the fetal left side, in the area of the lower chest and upper abdomen. The mass extended from at least the level of the umbilicus up to the neck but did not involve the neck. In the second case, prenatal sonography showed a multilocular, septate axillary cystic lymphangioma external to the chest with no flow on Doppler sonography. Follow-up sonography revealed normal fetal growth and enlargement of the cystic mass surrounding the left humerus. Both infants were delivered without complications and have been managed conservatively. PMID- 11425090 TI - Contrast-enhanced gray-scale and color Doppler voiding urosonography versus voiding cystourethrography in the diagnosis and grading of vesicoureteral reflux. AB - PURPOSE: The purpose of this study was to compare contrast-enhanced gray-scale voiding urosonography (CE-VUS) and contrast-enhanced color Doppler voiding urosonography (CE-CDVUS) with voiding cystourethrography (VCUG) to verify whether the use of color Doppler imaging improves the diagnosis and grading of vesicoureteral reflux (VUR). METHODS: In 74 patients, CE-VUS and CE-CDVUS were compared with VCUG, which was used as the gold standard. SHU 508 A (Levovist) was used as the echo-enhancing contrast agent. VUR was diagnosed if hyperechoic dots or color signals were visualized in the ureter on sonograms. VUR grading was based on morphologic and dynamic findings on CE-VUS and morphologic and color findings on CE-CDVUS. VCUG was performed conventionally, and grading by VCUG was in accordance with the international system of radiographic VUR grading. Patients who voided during 1 examination only (either CE-VUS and CE-CDVUS or VCUG) were excluded from the study. Agreement between the results of CE-VUS and VCUG and between those of CE-CDVUS and VCUG in diagnosing VUR was calculated by kappa statistics. CE-VUS and CE-CDVUS were compared for diagnostic accuracy by the McNemar test. RESULTS: The agreement between CE-VUS and VCUG in predicting VUR was 90% (kappa score, 0.77; p < 0.001). The agreement between CE-CDVUS and VCUG was 96% (kappa score, 0.91; p < 0.001). CE-CDVUS showed a significantly higher diagnostic accuracy than did CE-VUS (96% versus 90% of cases correctly classified; McNemar chi2 = 4; p < 0.05). This was mainly related to the lower number of false-negative results for grade I and grade II VUR when CE-CDVUS was used. CONCLUSIONS: The use of color Doppler imaging significantly improves the accuracy of contrast voiding urosonography in the detection and grading of VUR. PMID- 11425091 TI - Prenatal cocaine exposure and cranial sonographic findings in preterm infants. AB - PURPOSE: Prenatal cocaine exposure has been linked with subependymal hemorrhage and the formation of cysts that are detectable on cranial sonography in neonates born at term. We sought to determine if prenatal cocaine exposure increases the incidence of subependymal cysts in preterm infants. METHODS: We retrospectively reviewed the medical records and cranial sonograms obtained during a 1-year period on 122 premature (< 36 weeks of gestation) infants. Infants were categorized into 1 of 2 groups: those exposed to cocaine and those not exposed to cocaine. Infants were assigned to the cocaine-exposed group if there was a maternal history of cocaine abuse during pregnancy or if maternal or neonatal urine toxicology results were positive at the time of delivery. RESULTS: Five of the 122 infants were excluded from the study because of insufficient medical and drug histories. The incidence of subependymal cysts in the 117 remaining infants was 14% (16 of 117). The incidence of subependymal cysts in infants exposed to cocaine prenatally was 44% (8 of 18) compared with 8% (8 of 99) in the unexposed group (p < 0.01). CONCLUSIONS: We found an increased incidence of subependymal cyst formation in preterm infants who were exposed to cocaine prenatally. This result is consistent with results of similar studies in term infants. PMID- 11425092 TI - Value of chest sonography in the diagnosis and management of acute chest disease. AB - PURPOSE: The aim of this study was to investigate the value of chest sonography in the diagnosis and management of patients with chest radiograph opacities in an emergency department. METHODS: Seventy-eight patients with acute chest complaints whose chest radiographs showed opacities underwent chest sonography. The initial diagnosis (based on clinical manifestations and the chest radiograph), the sonographic diagnosis (before any invasive procedures), and the final diagnosis were compared. The impact of chest sonography on the management of patients with chest opacities was also analyzed. RESULTS: The initial diagnosis was in concordance with the final diagnosis in 60 (77%) of the 78 patients, while the sonographic diagnosis was in concordance with the final diagnosis in 75 (96%) of the patients. Chest sonography therefore significantly increased the rate of correct diagnoses from 77% (95% confidence interval, 67-87%) to 96% (95% confidence interval, 92-100%; p < 0.0001). Sonography provided new information in 52 patients (67%): a different diagnosis from the initial diagnosis in 18 patients and additional diagnostic information in 34 patients. New information gained from sonography affected the management of 35 patients. Sonography also provided help in guiding 42 (70%) of 60 invasive diagnostic procedures and 22 (73%) of 30 invasive therapeutic procedures for which chest radiography and physical examination had failed to or were unsuitable to provide guidance. Overall, sonography assisted in the management of 64 (82%) of 78 patients. Sonography was of no benefit in 14 patients (18%). CONCLUSIONS: We conclude that chest sonography can complement chest radiography and is of value in the diagnosis and management of emergency department patients with acute chest diseases presenting as opacities on chest radiographs. PMID- 11425093 TI - Correlation of right atrial pressure and blood flow velocities in the common femoral vein obtained by duplex Doppler sonography. AB - PURPOSE: The purpose of this study was to correlate the right atrial pressure with the blood flow velocity and pulsatility index (PI) values obtained with duplex Doppler sonography of the common femoral vein. METHODS: Thirty consecutive patients with suspected right-sided heart failure who had right atrial pressure measurements for correlation were included in the study. The common femoral veins were examined with duplex Doppler sonography. Irrespective of the presence or absence of clinical right-sided heart failure, patients were divided into 2 groups on the basis of their right atrial pressures. The mean flow velocities and PIs in the common femoral veins of the 2 groups were compared. RESULTS: In patients with elevated right atrial pressure (> or = 8 mm Hg), the mean minimum flow velocity + standard deviation (-0.15 +/- 0.11 m/second) in the common femoral vein was significantly lower than that in patients with normal right atrial pressure (< 8 mm Hg) (0.01 +/- 0.10 m/second) (p < 0.001). There was no significant difference in the mean maximum flow velocities in the common femoral vein between the patients with elevated right atrial pressure (0.25 +/- 0.08 m/second) and the patients with normal right atrial pressure (0.21 +/- 0.09 m/second). The mean PI in patients with elevated right atrial pressure (7.75 +/- 3.19) was significantly higher than the mean PI in patients with normal right atrial pressure (1.55 +/- 1.30; p < 0.001). There was a weak negative correlation between the minimum flow velocity and the pressure in the right atrium in patients with elevated right atrial pressure (p = 0.05; r = -0.4760). The amplitude of retrograde waves seen in patients with normal right atrial pressure was significantly smaller than in patients with elevated right atrial pressure (p < 0.05). CONCLUSIONS: The association of a decreased minimum velocity, especially a negative value, and an increase in PI in the common femoral vein may indicate an elevated right atrial pressure. Sonographic assessment of retrograde flow velocity in the common femoral vein may be useful in monitoring the response to medical treatment to decrease atrial pressure. PMID- 11425094 TI - Utility of lower-extremity duplex sonography in patients with venous thromboembolism. AB - PURPOSE: We evaluated the utility of venous duplex ultrasonography (VDUS) of the lower extremities in patients with pulmonary embolism (PE) and studied the distribution of venous thrombi in deep vein thrombosis (DVT) patients with and without PE. METHODS: We retrospectively reviewed medical records of all inpatients with a final diagnosis of PE or DVT between 1989 and 2000. RESULTS: Venous thrombosis was detected by VDUS in 229 patients (191 without PE and 38 with PE). The left leg only was involved in 50% of patients (p < 0.05), the right leg only in 33%, and both legs in 17%. The overall distribution of veins affected by DVT was: popliteal vein, 77%; superficial femoral vein, 76%; common femoral vein, 65%; posterior tibial vein, 23%; external iliac vein, 21%; common iliac vein, 9%; great saphenous vein, 7%; and inferior vena cava, 2%. A single venous site was involved in 22% of patients. External iliac vein thrombosis was more frequent in patients with DVT only (24%) than with PE and DVT (5%) (p < 0.05). The venous obstruction was partial in 14% of patients. VDUS of the asymptomatic leg was positive in 14% of patients with unilateral symptoms of DVT, all of whom also had DVT in the symptomatic leg. VDUS was positive for DVT in 90% of patients with PE and concomitant pain or edema of the leg, compared to only 20% of PE patients with no symptoms of DVT. CONCLUSIONS: Sonography should be the first diagnostic test for patients suspected of having PE with any sign or symptom related to DVT. VDUS of the asymptomatic leg is unnecessary in the diagnosis and management of DVT. Omitting the superficial femoral vein examination would lead to some decrease in the sensitivity of VDUS. PMID- 11425095 TI - Sonographic findings in solitary schwannoma of the breast. AB - Solitary schwannoma is a benign neoplasm of Schwann cell origin, most frequently occurring in the extremities and the trunk and head regions. The breast is a very unusual site for this tumor. We report a case of benign schwannoma of the breast in a 76-year-old woman. The palpable mass was localized in the lower outer quadrant of the right breast. This intramammary schwannoma appeared sonographically as a well-circumscribed, hypoechoic, slightly inhomogeneous solid mass. Microscopically, it had a pattern similar to that of schwannoma at other sites. PMID- 11425096 TI - Acoustical cavity excitation. AB - A tutorial exposition is given of cavity excitations and it is shown that relatively simple analytical solutions for the excitation of acoustic fields in cavities with lossy walls can be obtained for a broad class of such cavities. The standard analytic procedure whereby one expresses the acoustic field as a sum over the eigenmodes of the cavity is reviewed and its limitations are pointed out. An alternate procedure is to seek to make use of Green's functions depending on a single coordinate and to not express such Green's functions as sums over eigenfunctions. Instead, one expresses the Green's function as a closed form function that has a discontinuous slope at the value of the coordinate corresponding to the source location. Although the class of cavities for which this approach is possible is limited, it is sufficiently broad to be of intrinsic interest and the results may be helpful to those seeking benchmark solutions and analytic insight. Displayed examples are for cavities whose shapes are spheres and cylinders, with interior monopole, dipole, line, and distributed sources. The formulation yields simple expressions for the total power absorbed by the cavity walls. PMID- 11425098 TI - Pulse propagation in randomly fluctuating media. AB - Pulse propagation in a weakly and randomly inhomogeneous medium is studied using a time-domain progressive wave equation. An eikonal-like approximated solution to the wave equation derived from the path integral representation is used to obtain the time-dependent statistics of pulses propagating through this random medium. This approach yields both a simple way of producing simulations of time series as well as their statistical properties. PMID- 11425097 TI - A staggered-grid finite-difference method with perfectly matched layers for poroelastic wave equations. AB - A particle velocity-strain, finite-difference (FD) method with a perfectly matched layer (PML) absorbing boundary condition is developed for the simulation of elastic wave propagation in multidimensional heterogeneous poroelastic media. Instead of the widely used second-order differential equations, a first-order hyperbolic leap-frog system is obtained from Biot's equations. To achieve a high accuracy, the first-order hyperbolic system is discretized on a staggered grid both in time and space. The perfectly matched layer is used at the computational edge to absorb the outgoing waves. The performance of the PML is investigated by calculating the reflection from the boundary. The numerical method is validated by analytical solutions. This FD algorithm is used to study the interaction of elastic waves with a buried land mine. Three cases are simulated for a mine-like object buried in "sand," in purely dry "sand" and in "mud." The results show that the wave responses are significantly different in these cases. The target can be detected by using acoustic measurements after processing. PMID- 11425099 TI - Multiple scattering in a reflecting cavity: application to fish counting in a tank. AB - Classical fisheries acoustics techniques are useless in the presence of multiple scattering or reflecting boundaries. A general technique is developed that provides the number and the scattering strength of scatterers in motion placed inside a highly reflecting cavity. This approach is based on multiple scattering theory. The idea is to measure the average effect of the scatterers on the acoustic echoes of the cavity interfaces. This leads to the measure of the scattering mean free path, a typical length that characterizes the scattering strength of the cloud of scatterers. Numerical results are shown to agree with a simple theoretical analysis. Experiments are performed with fish in a tank at two different scales: ultrasonic frequency (400 kHz) in a 1.4-l beaker with 1-cm-long fish as well as fisheries acoustics frequency (12.8 kHz) in a 30-m3 tank with 35 cm-long fish. These results have interesting applications to fish target strength measurement and fish counting in aquaculture. PMID- 11425100 TI - Reflectance properties of two-dimensional sonic band-gap crystals. AB - An analysis of the reflectance of sonic band-gap crystals consisting of square arrays of rigid cylinders in air is presented. The standing wave formed in front of the structures is studied both experimentally and theoretically. Experiments have been performed with a mobile robotized microphone that obtains pressure maps on the plane perpendicular to the axes of the cylinders. Enhancements of the standing wave ratio (SWR) are observed in frequency regions where attenuation bands appear in zero-order transmission experiments. Also, the SWR presents oscillations that can be related to the finite dimension of the structure (Fabry Perot effect). Both features are well described by calculations based on a double scattering approach. PMID- 11425101 TI - Acoustically coupled gas bubbles in fluids: time-domain phenomena. AB - In previous work [C. Feuillade, J. Acoust. Soc. Am. 98, 1178-1190 (1995)] a coupled oscillator formalism was introduced for describing collective resonances, scattering, and superresonances, of multiple gas bubbles in a fluid. Subsequently, time-domain investigations of the impulse response of coupled systems have disclosed the exact conditions which determine whether the ensemble scattering behavior should be described using: either (a), a multiple scattering; or (b), a self-consistent methodology. The determining factor is the Q of the individual scatterers, and their typical spatial separations in the medium. For highly damped or sparse systems, e.g., scattering from loose schools of swimbladder fish, or from a gassy seabed containing entrained bubbles, the multiple scatter counting approach should be applicable. For more strongly coupled systems, e.g., a dense cloud of resonating bubbles in the water column, energy exchange may be due primarily to radiative cycling rather than scattering, in which case a self-consistent approach is indicated. The result has implications for both volume and bottom scattering applications. PMID- 11425102 TI - Time reversal for a single spherical scatterer. AB - We show that the time reversal operator for a planar time reversal mirror (TRM) can have up to four distinct eigenvalues with a small spherical acoustic scatterer. Each eigenstate represents a resonance between the TRM and an induced scattering moment of the sphere. Their amplitude distributions on the TRM are orthogonal superpositions of the radiation patterns from a monopole and up to three orthogonal dipoles. The induced monopole moment is associated with the compressibility contrast between the sphere and the medium, while the dipole moments are associated with density contrast. The number of eigenstates is related to the number of orthogonal orientations of each induced multipole. For hard spheres (glass, metals) the contribution of the monopole moment to the eigenvalues is much greater than that of the dipole moments, leading to a single dominant eigenvalue. The other eigenvalues are much smaller, making it unlikely multiple eigenvalues could have been observed in previous experiments using hard materials. However, for soft materials such as wood, plastic, or air bubbles the eigenvalues are comparable in magnitude and should be observable. The presence of multiple eigenstates breaks the one-to-one correspondence between eigenstates and distinguishable scatterers discussed previously by Prada and Fink [Wave Motion 20, 151-163 (1994)]. However, eigenfunctions from separate scatterers would have different phases for their eigenfunctions, potentially restoring the ability to distinguish separate scatterers. Since relative magnitudes of the eigenvalues for a single scatterer are governed by the ratio of the compressibility contrast to the density contrast, measurement of the eigenvalue spectrum would provide information on the composition of the scatterer. PMID- 11425103 TI - The surface motion due to a line force or dislocation within an anisotropic elastic half-space. AB - An explicit solution of the surface displacements due to a line force or a line dislocation within an anisotropic half-space is presented. The surface displacements are derived from the solution corresponding to a suddenly applied surface line force using the reciprocal theorem. The solution is in a closed form for isotropic media. For anisotropic solids, only an eigenvalue problem needs to be solved numerically for a given time and position to calculate the surface response. Numerical results are given for silicon. PMID- 11425104 TI - Modeling elastic wave forward propagation and reflection using the complex screen method. AB - Formulation for calculating forward propagation and reflection in a 3D elastic structure based on the complex-screen method is given in this paper. The calculation of reflections is formulated based on the local Born approximation. When using a small angle approximation, the backscattering operator reduces to a screen operator which is similar to the forward screen propagator. Combining the forward propagator and backscattering operator together, the new method can properly handle the multiple forward scattering and single backscattering in a 3D heterogeneous model. Using a dual-domain technique, the new method is highly efficient in CPU time and memory savings. For models where reverberation and resonance scattering can be neglected, this method provides a fast and accurate algorithm. Synthetic seismograms for two-dimensional elastic models are calculated with this method and compared with those generated by the finite difference method. The results show that the method works well for small to medium scattering angles and medium velocity contrasts. PMID- 11425105 TI - Mode counts in an aluminum foam. AB - Measurements of the ultrasonic modal density of a disordered elastic frame, a 20 pore-per-inch open-celled aluminum foam, are reported. While the material is dissipative, with a Q only around 700, sufficiently careful signal processing has allowed reliable counts of the modes up through a few hundred, corresponding to wavelengths comparable to the strut lengths. The modal density is found to be essentially constant over this range, and to bear no resemblance to theoretical estimates based on long-wavelength effective moduli. PMID- 11425106 TI - Acoustic nonlinearity of cracks partially filled with liquid: cubic approximation. AB - The theoretical investigation of mechanisms of the acoustic nonlinearity (elastic and inelastic) of cracks partially filled with an ideal and viscous liquid and associated with the nonlinear dependence of the capillary and viscous pressure in the liquid on the distance between the crack surfaces and the velocity of the change of this distance is proposed. The nonlinear (in cubic approximation) equations of the state of these cracks is obtained, and its parameters are defined. It is shown that the presence of the viscous liquid may lead to the considerable increase of the acoustic nonlinearity of such cracks in comparison with cracks filled with the ideal liquid. PMID- 11425107 TI - Weak nonlinear propagation of sound in a finite exponential horn. AB - This article presents an approximate solution for weak nonlinear standing waves in the interior of an exponential acoustic horn. An analytical approach is chosen assuming one-dimensional plane-wave propagation in a lossless fluid within an exponential horn. The model developed for the propagation of finite-amplitude waves includes linear reflections at the throat and at the mouth of the horn, and neglects boundary layer effects. Starting from the one-dimensional continuity and momentum equations and an isentropic pressure-density relation in Eulerian coordinates, a perturbation analysis is used to obtain a hierarchy of wave equations with nonlinear source terms. Green's theorem is used to obtain a formal solution of the inhomogeneous equation which takes into account linear reflections at the ends of the horn, and the solution is applied to the nonlinear horn problem to yield the acoustic pressure for each order, first in the frequency and then in the time domain. In order to validate the model, an experimental setup for measuring fundamental and second harmonic pressures inside the horn has been developed. For an imposed throat fundamental level, good agreement is obtained between predicted and measured levels (fundamental and second harmonic) at the mouth of the horn. PMID- 11425108 TI - Numerical model for nonlinear standing waves and weak shocks in thermoviscous fluids. AB - Nonlinear standing waves in a one-dimensional tube are studied numerically by using a finite-difference algorithm. The numerical code models the acoustic field in resonators for homogeneous, thermoviscous fluids. Calculations are performed exclusively in the time domain, and all harmonic components are obtained by one resolution. The fully nonlinear differential equation is written in Lagrangian coordinates. It is solved without truncation. Effects of absorption are included. Displacement and pressure wave forms are calculated at different locations and results are shown for different excitation levels and tube lengths. Amplitude distributions along the resonator axis for every harmonic component are also evaluated. Simulations are performed for amplitudes ranging from linear to strongly nonlinear and weak shock. A very good concordance with classic experimental and analytical results is obtained. PMID- 11425109 TI - Time-reversed sound beams of finite amplitude. AB - Numerical simulations based on the nonlinear parabolic wave equation are used to investigate time reversal of sound beams radiated by unfocused and focused sources. Emphasis is placed on nonlinear propagation distortion in the time reversed beam, and specifically its effect on field reconstruction. Distortion of this kind, due to amplification during time reversal, has been observed in recent experiments [A. P. Brysev et al., Acoust. Phys. 44, 641-650 (1998)]. Effects of diffraction introduced by time-reversal mirrors with finite apertures are also considered. It is shown that even in the presence of shock formation, the ability of time reversal to retarget most of the energy on the source or focal region of the incident beam is quite robust. PMID- 11425110 TI - Blast noise propagation above a snow cover. AB - A porous medium model of a snow cover, rather than a viscoelastic treatment, has been used to simulate measured, horizontally traveling acoustic waveform propagation above a dry snow cover 11-20 cm thick. The waveforms were produced by explosions of 1-kg charges at propagation distances of 100 to 1400 m. These waveforms, with a peak frequency around 30 Hz, show pulse broadening effects similar to those previously seen for higher-frequency waves over shorter propagation distances. A rigid-ice-frame porous medium ("rigid-porous") impedance model, which includes the effect of the pores within the snow but ignores any induced motion of the ice particles, is shown to produce much better agreement with the measured waveforms compared with a viscoelastic solid treatment of the snow cover. From the acoustic waveform modeling, the predicted average snow cover depth of 18 cm and effective flow resistivities of 16-31 kPa s m(-2) agree with snow pit observations and with previous acoustic measurements over snow. For propagation in the upwind direction, the pulse broadening caused by the snow cover interaction is lessened, but the overall amplitude decay is greater because of refraction of the blast waves. PMID- 11425111 TI - Acoustic sounding of wind velocity profiles in a stratified moving atmosphere. AB - The paper deals with analytical and numerical studies of the effects of atmospheric stratification on acoustic remote sensing of wind velocity profiles by sodars. Both bistatic and monostatic schemes are considered. Formulas for the Doppler shift of an acoustic echo signal scattered by atmospheric turbulence advected with the mean wind in a stratified moving atmosphere are derived. Numerical studies of these formulas show that errors in retrieving wind velocity can be of the order of 1 m/s if atmospheric stratification is ignored. Formulas for the height at which wind velocity is retrieved are also derived. Approaches are proposed which allow one to take into account the effects of atmospheric stratification when restoring the wind velocity profile from measured values of the Doppler shift and the time interval of acoustic impulse propagation from a sodar to the scattering volume and back to the ground. PMID- 11425112 TI - On the stability of long-range sound propagation through a structured ocean. AB - Several acoustic experiments show a surprising degree of stability in wave fronts propagating over multi-megameter ranges through the ocean's sound channel despite the presence of random-like, sound-speed fluctuations. Previous works have pointed out the existence of chaos in simplified ray models incorporating structure inspired by the true ocean environment. A "predictability horizon" has been introduced beyond which stable wavefronts cease to exist and point-wise, detailed comparisons between even the most sophisticated models and experiment may be limited for fundamental reasons. By applying one of the simplified models it is found that, for finite ranges, the fluctuations of the ray stabilities are very broad and consistent with log-normal densities. A fraction of the ray density retains a much more stable character than the typical ray. This may be one of several possible mechanisms leading to greater than anticipated sound field stability. The log-normal ray stability density may underlie the recent, experimentally determined, log-normal density of wave-field intensities [Colosi et al., J. Acoust. Soc. Am. 105, 3202-3218 (1999)]. PMID- 11425113 TI - Three-dimensional elasticity model for a decoupling coating on a rectangular plate immersed in a heavy fluid. AB - This paper presents an analysis of the vibroacoustic response of a finite, simply supported rectangular plate covered by a layer of decoupling material and immersed in a heavy fluid. An exact formulation using the three-dimensional theory of elasticity for the decoupling material is derived for this problem, thereby extending previous studies that were limited to infinite plates. The paper details the constitutive equations of the problem and the analytical method of solution. Numerical results show that shear waves in the decoupling material generally have little influence on the sound radiation in the heavy fluid. Comparisons with a locally reacting model of the decoupling material and with the simple model of House [Proc. I.O.A. 13(3), 166-173 (1991)] are also presented. PMID- 11425114 TI - A modal Pritchard approximation for computing array element mutual impedance. AB - An investigation into the applicability and accuracy of Pritchard's approximation for closely packed transducer arrays is undertaken. A new, "modal" Pritchard approximation is developed, based upon normal modes of the acoustic medium, and is tested for arrays of acoustically hard spheres to ascertain its accuracy in determining the mutual acoustic radiation impedance between array elements. For ka approximately 1, it is found that the modal Pritchard approximation works quite well in approximating the mutual radiation impedance of a two-element array, even for relatively close spacing; but for arrays of three or more scatterers in close proximity the approximation may have relatively large errors. The effect of neglecting inter-element scattering is analyzed for the monopole-to monopole scattering of various configurations of a three-element array and a sixteen-element double line array. PMID- 11425115 TI - A multifrequency scintillation method for ocean flow measurement. AB - The transverse flow of inhomogeneous fluid produces fluctuation of the acoustic signal passing through it. The coherence of frequency-spaced signal fluctuation is related to the advection of the inhomogeneous medium through the sound path, thus providing a basis for the current velocity measurement. This method can be considered to be the "frequency-domain" version of the conventional scintillation approach to the current velocity registration based on the measurement of the signal correlation transmitted from the source to the two separated in space receivers (space-domain scintillation) [S. Clifford and D. Farmer, J. Acoust. Soc. Am. 74, 1826-1832 (1983)]. The sensitivity of the method depends on the features of the ocean fine structure, which is determined mainly by the internal waves and turbulence. To estimate the sensitivity of the multifrequency method of transverse current probing, the coherence function of two signals propagating through a frozen and moving internal wave field and through the turbulence is considered. The application of the multifrequency signal allows estimation of the fine-structure parameters as well as the current velocity. PMID- 11425116 TI - Stationary velocity and pressure gradients in a thermoacoustic stack. AB - The second-order time-averaged acoustics of a viscous, thermally conducting gas between closely spaced parallel plates is studied. The acoustic disturbance is studied by expanding the equations of fluid dynamics and heat transfer to second order in Mach number. The undisturbed state is allowed to have a nonzero temperature gradient. A set of coupled equations for the time-averaged pressure gradient, velocity, and temperature are obtained and solved. Particular attention is paid to the relation between the time-averaged mass flux and pressure gradient. An explicit expression is obtained relating the time-averaged pressure drop across a thermoacoustic stack to the time-averaged mass flux through the stack. PMID- 11425117 TI - Development of panel loudspeaker system: design, evaluation and enhancement. AB - Panel speakers are investigated in terms of structural vibration and acoustic radiation. A panel speaker primarily consists of a panel and an inertia exciter. Contrary to conventional speakers, flexural resonance is encouraged such that the panel vibrates as randomly as possible. Simulation tools are developed to facilitate system integration of panel speakers. In particular, electro mechanical analogy, finite element analysis, and fast Fourier transform are employed to predict panel vibration and the acoustic radiation. Design procedures are also summarized. In order to compare the panel speakers with the conventional speakers, experimental investigations were undertaken to evaluate frequency response, directional response, sensitivity, efficiency, and harmonic distortion of both speakers. The results revealed that the panel speakers suffered from a problem of sensitivity and efficiency. To alleviate the problem, a woofer using electronic compensation based on H2 model matching principle is utilized to supplement the bass response. As indicated in the result, significant improvement over the panel speaker alone was achieved by using the combined panel-woofer system. PMID- 11425118 TI - Time-domain analyses of acoustics-structure interactions for piezoelectric transducers. AB - Piezoelectric transducers coupled with a surrounding medium are analyzed in time domain using the coupled finite element and boundary element method. Three dimensional solid elements are employed to model the piezoelectric transducer, while the surrounding medium is described by the boundary integral equation and the boundary of the medium is modeled by two-dimensional spatial elements. Verification studies were conducted to evaluate the accuracy and convergence of the present numerical algorithm and they show that the present numerical solutions agree well with the analytical ones. The influence of the surrounding medium on the acoustic field is studied. The interaction between the structure and surrounding medium affects the structure dynamic performances and acoustic pressure distributions significantly. However, in the present study we show that the radiation directivity is insignificantly influenced by the surrounding medium. PMID- 11425119 TI - Visualization of acoustic radiation from a vibrating bowling ball. AB - This paper presents visualization of acoustic radiation from a vibrating bowling ball using the Helmholtz equation least squares (HELS) method. In conducting the experiments, the ball is excited by a vibration shaker using stationary random signals. The radiated acoustic pressures are measured using two microphones and taken as input to the HELS formulations. The reconstructed acoustic pressures on the bowling ball surface are compared with those measured at the same locations. Also shown are comparisons of the reconstructed and measured acoustic pressure spectra at various locations on the bowling ball surface. Results demonstrate that the accuracy of reconstruction based on measurements over a conformal surface is much higher than that over a finite planar surface. This is because the latter often extends beyond the near-field region, making the accuracy of measurements inconsistent. Nevertheless, satisfactory reconstruction of acoustic pressure fields over the entire bowling ball surface can still be obtained based on the measurements taken over a finite planar surface on one side of the source. In a similar manner, the normal component of the surface velocity is reconstructed. Once these acoustic quantities are determined, the time-averaged acoustic intensity is calculated. Also presented are the formulations for estimating a priori the numbers of expansion functions and measurements required by the HELS method and the guidelines for determining the reconstruction error and optimum measurement locations, given the overall dimensions of the source and the highest frequency of interest in reconstruction. PMID- 11425120 TI - Experimental and numerical investigations of resonant vibration characteristics for piezoceramic plates. AB - Electronic speckle pattern interferometry (ESPI) is a full field, non-contact technique for measuring the surface displacement of a structure subjected to static loading or, especially, to dynamic vibration. In this article we employ an optical system called the amplitude-fluctuation ESPI with out-of-plane and in plane measurements to investigate the vibration characteristics of piezoceramic plates. Two different configurations of piezoceramic plates, namely the rectangular and the circular plates, are discussed in detail. As compared with the film recording and optical reconstruction procedures used for holographic interferometry, the interferometric fringes of AF-ESPI are produced instantly by a video recording system. Because the clear fringe patterns will be shown only at resonant frequencies, both the resonant frequencies and the corresponding mode shapes are obtained experimentally at the same time by the proposed AF-ESPI method. Excellent quality of the interferometric fringe patterns for both the in plane and out-of-plane vibration mode shapes is demonstrated. The resonant frequencies of the piezoceramic plates are also measured by the conventional impedance analysis. From experimental results, we find that the out-of-plane vibration modes (type A) with lower resonant frequencies cannot be measured by the impedance analysis and only the in-plane vibration modes (type B) will be shown. However, both the out-of-plane (bending) and in-plane (extensional) vibration modes of piezoceramic plates are obtained by the AF-ESPI method. Finally, the numerical finite element calculations are also performed, and the results are compared with the experimental measurements. It is shown that the numerical calculations and the experimental results agree fairly well for both the resonant frequencies and the mode shapes. PMID- 11425121 TI - Transient response of an acoustic medium by an excited submerged spherical shell. AB - An exact closed-form solution is obtained for the transient response of an acoustic fluid due to an excited submerged spherical shell. Step axisymmetric stress acting on the inner surface of the shell is expanded into modal Fourier components in which for each modal stress systematic exact expressions for modal radial displacement of the shell, fluid pressure, fluid particle velocity and displacement are presented. The superiority of the formulations herein is its applicability to any time and spatial distance in the fluid as well as on the shell. The formulation also does not require additional numerical computations associated with other methods. PMID- 11425122 TI - The surface variational principle applied to an acoustic cavity. AB - This paper presents the development and application of the Surface Variational Principle (SVP) for the evaluation of axisymmetric interior acoustic domains. The interior form of the SVP is first developed in the same manner as the existing exterior form. Then, the surface pressure and normal velocity are represented with a Ritz expansion using basis functions that span the entire wetted surface of the object of interest. The resultant formulation is used to analyze the interior acoustic response of a harmonically forced, right circular elastic cylinder. This validation model was chosen as both the structural and acoustic responses can be solved analytically. Results are presented for two models: one with a length to radius ratio of 2.4, and another with a ratio of 12.3. The SVP is shown to well reproduce the analytical solution for this geometry, and displays the asymptotic convergence expected of its variational formulation. The SVP formulation developed here is not restricted to right-circular cylindrical geometries, and may, indeed, be readily applied to any axisymmetric body. PMID- 11425123 TI - A theoretical study of passive control of duct noise using panels of varying compliance. AB - It is theoretically demonstrated that, in a duct, a substantial amount of sound energy can be transferred to flexural waves on a finite wall panel when the upstream portion of the panel is made to couple strongly with sound. The flexural wave then loses its energy either through radiating reflection sound waves or by internal friction. The effectiveness of the energy transfer and damping is greatly enhanced if the panel has a gradually decreasing in vacuo wave speed, which, in this study, is achieved by using a tapered membrane under tension. A high noise attenuation rate is possible with the usual viscoelastic materials such as rubber. The transmission loss has a broadband spectrum, and it offers an alternative to conventional duct lining where a smooth air passage is desired and nonacoustical considerations, such as chemical contamination or cost of operation maintenance, are important. Another advantage of the tapered panel is that, at very low frequencies, typically 5% of the first cut-on frequency of the duct, sound reflection occurs over the entire panel length. This supplements the inevitable drop in sound absorption coefficient, and a high transmission loss may still be obtained at very low frequencies. PMID- 11425124 TI - A study of time-domain FXLMS algorithms with control output constraint. AB - A single input, single output active noise control system using the time-domain Filtered-X LMS algorithm with output constraint is investigated. The constraint on the output of the control filter is applied by three different methods: the leakage algorithm based on the transformation method using a penalty function; the re-scaling algorithm based on the active set method; and the simple practical (clipping) algorithm which just clips the output if a constraint is encountered. A comparison of the three algorithms shows that the re-scaling algorithm can usually work successfully under the constraint, while the leakage algorithm usually needs a large leakage coefficient to satisfy the constraint with a resulting performance loss. The clipping algorithm has potential problems both with the stability and convergence speed. PMID- 11425125 TI - Orthogonal acoustical factors of a sound field in a bamboo forest. AB - To investigate the acoustical quality of a sound field in a bamboo forest, acoustical measurements were conducted to obtain orthogonal acoustical factors of the sound field. These results are compared with previous results for a sound field in an ordinary forest [H. Sakai, S. Sato, and Y. Ando, J. Acoust. Soc. Am. 104, 1491-1497 (1998)]. The IACC, which is defined as a maximum value of the normalized interaural cross-correlation function between signals at the ears, was 0.07 (4 kHz) and 0.16 (2 kHz) at positions 20 and 40 m from the source, respectively. These values are much better than those in the previously investigated forest. The measured subsequent reverberation time Tsub was up to 1.5 s in the frequency range above 1 kHz at the position 40 m from the source. For certain music sources with higher frequency components, therefore, sound fields in a bamboo forest have excellent acoustic properties. PMID- 11425126 TI - Minimum entropy deconvolution of pulse-echo signals acquired from attenuative layered media. AB - In this article deconvolution of ultrasonic pulse-echo data acquired from attenuative layered media is considered. The problem is divided in two subproblems: treating the sparse reflection sequence caused by the layered structure of the media and treating the frequency-dependent attenuation. The first subproblem is solved by means of joint maximum a posteriori estimation of the assumed zero mean, white, nonstationary reflection sequence and its corresponding sequence of unknown standard deviations. This approach leads to an algorithm that seeks minimum entropy solutions for the reflection sequence and therefore the algorithm serves as a novel link between the classical Wiener filter and methods for sparse or minimum entropy deconvolution. The second subproblem is solved by introducing a new signal processing-oriented, linear discrete-time model for frequency-dependent attenuation in isotropic and homogeneous media. The deconvolution algorithm is tested using simulated data and its performance for real normal incidence pulse-echo data from a composite material is also demonstrated. The results show that the algorithm, in combination with the attenuation model, yields estimates that reveal the internal structure of the composite and, thus, simplify the interpretation of the ultrasonic data. PMID- 11425127 TI - A model for sound lateralization. AB - Recent studies of multiple sclerosis (MS) and stroke patients suggested a correlation between two patterns of abnormal performance in lateralization tasks and two sites of pontine lesions. Most patients who had lesions below or at the superior olivary complex (SOC) perceived all interaural differences in binaural stimuli as small, while most patients who had lesions above the SOC perceived all interaural differences as large. The two abnormal performance patterns occurred for interaural time differences (ITD) and/or for interaural level differences (ILD). The present model proposes a multi-level hierarchical brainstem structure that estimates ITD and ILD. The first level seeks dissimilarity between the left and right inputs and a second level looks for similarity between the two sides' inputs. Each level is modeled as an ensemble of neural arrays in which each unit performs a logic or arithmetic function. The inputs are simulations of auditory nerve responses to broadband stimuli. Simulations yield good correspondence to the effect of both locations of pontine lesions on binaural performance. PMID- 11425128 TI - A computational algorithm for computing nonlinear auditory frequency selectivity. AB - Computational algorithms that mimic the response of the basilar membrane must be capable of reproducing a range of complex features that are characteristic of the animal observations. These include complex input output functions that are nonlinear near the site's best frequency, but linear elsewhere. This nonlinearity is critical when using the output of the algorithm as the input to models of inner hair cell function and subsequent auditory-nerve models of low- and high spontaneous rate fibers. We present an algorithm that uses two processing units operating in parallel: one linear and the other compressively nonlinear. The output from the algorithm is the sum of the outputs of the linear and nonlinear processing units. Input to the algorithm is stapes motion and output represents basilar membrane motion. The algorithm is evaluated against published chinchilla and guinea pig observations of basilar membrane and Reissner's membrane motion made using laser velocimetry. The algorithm simulates both quantitatively and qualitatively, differences in input/output functions among three different sites along the cochlear partition. It also simulates quantitatively and qualitatively a range of phenomena including isovelocity functions, phase response, two-tone suppression, impulse response, and distortion products. The algorithm is potentially suitable for development as a bank of filters, for use in more comprehensive models of the peripheral auditory system. PMID- 11425129 TI - Sources of distortion product otoacoustic emissions revealed by suppression experiments and inverse fast Fourier transforms in normal ears. AB - Primary and secondary sources combine to produce the 2f1-f2 distortion product otoacoustic emission (DPOAE) measured in the ear canals of humans. DPOAEs were obtained in nine normal-hearing subjects using a fixed-f2 paradigm in which f1 was varied. The f2 was 2 or 4 kHz, and absolute and relative primary levels were varied. Data were obtained with and without a third tone (f3) placed 15.6 Hz below 2f1-f2. The level of f3 was varied in order to suppress the stimulus frequency otoacoustic emission (SFOAE) coming from the 2f1-f2 place. These data were converted from the complex frequency domain into an equivalent time representation using an inverse fast Fourier transform (IFFT). IFFTs of unsuppressed DPOAE data were characterized by two or more peaks. Relative amplitudes of these peaks depended on overall primary level and on primary-level differences. The suppressor eliminated later peaks, but early peaks remained relatively unaltered. Results are interpreted to mean that the DPOAE measured in humans includes components from the f2 place (intermodulation distortion) and DP place (in the form of a SFOAE). These findings build on previous work by providing evidence that multiple peaks in the IFFT are due to a secondary source at the DP place. PMID- 11425130 TI - Duration discrimination and subjective duration for ramped and damped sounds. AB - The perception of stimuli with ramped envelopes (gradual attack and abrupt decay) and damped envelopes (abrupt attack and gradual decay) was studied in subjective and objective tasks. Magnitude estimation (ME) of perceived duration was measured for broadband noise, 1.0-kHz, and 8.0-kHz tones for durations between 10 and 200 ms. Damped sounds were judged to be shorter than ramped sounds. Matching experiments between sounds with ramped, damped, and rectangular envelopes also showed that damped sounds are perceived to be shorter than ramped sounds, and, additionally, the reason for the effect is a result of the damped sound being judged shorter than a rectangular-gated sound rather than the ramped sound being judged longer than a rectangular-gated sound. These matching studies also demonstrate that the size of the effect is larger for tones (factor of 2.0) than for broadband noise (factor of 1.5). There are two plausible explanations for the finding that damped sounds are judged to be shorter than ramped or rectangular gated sounds: (1) the abrupt offset at a high level of the ramped sound (or a rectangular-gated sound) results in a persistence of perception (forward masking) that is considered in judgments of the subjective duration; and (2) listeners may ignore a portion of the decay of a damped sound because they consider it an "echo" [Stecker and Hafter, J. Acoust. Soc. Am. 107, 3358-3368 (2000)]. In another experiment, duration discrimination for broadband noise with ramped, damped, and rectangular envelopes was studied as a function of duration (10 to 100 ms) to determine if differences in perceived duration are associated with the size of measured Weber fractions. A forced-choice adaptive procedure was used. Duration discrimination was poorer for noise with ramped envelopes than for noise with damped or rectangular envelopes. This result is inconsistent with differences in perceived duration and no explanation was readily apparent. PMID- 11425131 TI - Children's detection of pure-tone signals with random multitone maskers. AB - Preschoolers and adults were asked to detect a 1000-Hz signal, which was masked by a multitone complex. The frequencies and amplitudes of the components in the complex varied randomly and independently on each presentation. A staircase, cued two-interval, forced-choice procedure disguised as a "listening game" was used to obtain signal thresholds in quiet and in the presence of the multitone maskers. The number of components in the masker was fixed within an experimental condition and varied from 2 to 906 across experimental conditions. Thresholds were also measured with a broadband noise masker. Eight preschool children and eight adults were tested. Although individual differences were large, among both adults and children, there was little difference between the groups in the mean amount of masking produced by the maskers with large numbers of components (400 and 906). There was also a small but significant difference between adults and children in the mean amount of masking produced by the broadband noise. The difference between the groups was much larger with smaller numbers of components. Data obtained from the adults were basically similar to that previously reported [cf. Neff and Green, Percept. Psychophys. 41, 409-415 (1987); Oh and Lutfi, J. Acoust. Soc. Am. 104, 3489-3499 (1998)]: maskers comprised of 10-40 components produced as much as 30 to 60 dB of masking in some, but not all listeners. Those same maskers produced larger amounts of masking (70-83 dB) in many of the preschool children, although, as in the adult group, individual differences were large. The component-relative-entropy (CoRE) model [Lutfi, J. Acoust. Soc. Am. 94, 748-758 (1993)] was used to describe the differences in performance between the children and adults. According to this model the average child appears to integrate information over a larger number of auditory filters than the average adult. PMID- 11425132 TI - Using statistical decision theory to predict speech intelligibility. I. Model structure. AB - This article introduces a new model that predicts speech intelligibility based on statistical decision theory. This model, which we call the speech recognition sensitivity (SRS) model, aims to predict speech-recognition performance from the long-term average speech spectrum, the masking excitation in the listener's ear, the linguistic entropy of the speech material, and the number of response alternatives available to the listener. A major difference between the SRS model and other models with similar aims, such as the articulation index, is this model's ability to account for synergetic and redundant interactions among spectral bands of speech. In the SRS model, linguistic entropy affects intelligibility by modifying the listener's identification sensitivity to the speech. The effect of the number of response alternatives on the test score is a direct consequence of the model structure. The SRS model also appears to predict the differential effect of linguistic entropy on filter condition and the interaction between linguistic entropy, signal-to-noise ratio, and language proficiency. PMID- 11425133 TI - Using statistical decision theory to predict speech intelligibility. II. Measurement and prediction of consonant-discrimination performance. AB - The speech recognition sensitivity (SRS) model [H. Musch and S. Buus, J. Acoust. Soc. Am. 109, 2896-2909 (2001)] was tested by applying it to consonant discrimination data collected in this study. Normally hearing listeners' abilities to discriminate among 18 consonants were measured in 58 filter conditions using two test paradigms. In one paradigm, listeners chose among all 18 stimuli. In the other, response alternatives were restricted to the correct response and eight consonants that were randomly selected among the 17 incorrect response alternatives. The effect of the number of response alternatives on performance can be described by statistical decision theory. Most filter conditions included one or more sharply filtered narrow bands of speech. Depending on the selection of bands, listeners' performance in multi-band conditions falls short of, equals, or exceeds the performance expected from multiplication of the error rates in the individual bands. The performance advantage in multi-band conditions increases with average band separation. The SRS model provides a good fit to the data and predicts the data more accurately than does the speech intelligibility index. PMID- 11425134 TI - Psychophysical recovery from single-pulse forward masking in electric hearing. AB - Psychophysical single-pulse forward-masking (SPFM) recovery functions were measured for three electrodes in each of eight subjects with the nucleus mini-22 cochlear implant. Masker and probe stimuli were single 200-micros/phase biphasic current pulses. Recovery functions were measured at several masker levels spanning the electric dynamic range of electrodes chosen from the apical, middle, and basal regions of each subject's electrode array. Recovery functions were described by an exponential process in which threshold shift (in microA) decreased exponentially with increasing time delay between the masker and probe pulses. Two recovery processes were observed: An initial, rapid-recovery process with an average time constant of 5.5 ms was complete by about 10 ms. A second, slow-recovery process involved less masking than the rapid-recovery process but encompassed much longer time delays, sometimes as long as several hundred milliseconds. Growth-of-masking slopes for the rapid process depended upon time delay, as expected in an exponential recovery process. Unity slopes were observed at a time delay of 0 ms, whereas progressively shallower slopes were observed at time delays of 2 ms and 5 ms. Many recovery functions demonstrated nonmonotonicities or "facilitation" at very short masker-probe delays (1-2 ms). Such nonmonotonicities were usually most pronounced at low masker levels. Time constants for the rapid-recovery process did not vary systematically with masker level or with electrode location along the implanted array. Most subjects demonstrated rapid-recovery time constants less than 7 ms; however, the subject with the longest duration of deafness prior to implantation exhibited clearly prolonged time constants (9-24 ms). Time constants obtained on basal electrodes were inversely related to word recognition scores. PMID- 11425135 TI - Effects of temporal envelope modulation on acoustic signal recognition in a vocal fish, the plainfin midshipman. AB - Amplitude modulation is an important parameter defining vertebrate acoustic communication signals. Nesting male plainfin midshipman fish, Porichthys notatus, emit simple, long duration hums in which modulation is strikingly absent. Envelope modulation is, however, introduced when the hums of adjacent males overlap to produce acoustic beats. Hums attract gravid females and can be mimicked with continuous tones at the fundamental frequency. While individual hums have flat envelopes, other midshipman signals are amplitude modulated. This study used one-choice playback tests with gravid females to examine the role of envelope modulation in hum recognition. Various pulse train and two-tone beat stimuli resembling natural communication signals were presented individually, and the responses compared to those for continuous pure tones. The effectiveness of pulse trains was graded and depended upon both pulse duration and the ratio of pulse to gap length. Midshipman were sensitive to beat modulations from 0.5 to 10 Hz, with fewer fish approaching the beat than the pure tone. Reducing the degree of modulation increased the effectiveness of beat stimuli. Hence, the lack of modulation in the midshipman's advertisement call corresponds to the importance of envelope modulation for the categorization of communication signals even in this relatively simple system. PMID- 11425136 TI - Pitch strength and pitch dominance of iterated rippled noises in hearing-impaired listeners. AB - Reports using a variety of psychophysical tasks indicate that pitch perception by hearing-impaired listeners may be abnormal, contributing to difficulties in understanding speech and enjoying music. Pitches of complex sounds may be weaker and more indistinct in the presence of cochlear damage, especially when frequency regions are affected that form the strongest basis for pitch perception in normal hearing listeners. In this study, the strength of the complex pitch generated by iterated rippled noise was assessed in normal-hearing and hearing-impaired listeners. Pitch strength was measured for broadband noises with spectral ripples generated by iteratively delaying a copy of a given noise and adding it back into the original. Octave-band-pass versions of these noises also were evaluated to assess frequency dominance regions for rippled-noise pitch. Hearing-impaired listeners demonstrated consistently weaker pitches in response to the rippled noises relative to pitch strength in normal-hearing listeners. However, in most cases, the frequency regions of pitch dominance, i.e., strongest pitch, were similar to those observed in normal-hearing listeners. Except where there exists a substantial sensitivity loss, contributions from normal pitch dominance regions associated with the strongest pitches may not be directly related to impaired spectral processing. It is suggested that the reduced strength of rippled-noise pitch in listeners with hearing loss results from impaired frequency resolution and possibly an associated deficit in temporal processing. PMID- 11425137 TI - Aging and temporal discrimination in auditory sequences. AB - This study examined age-related changes in temporal sensitivity to increments in the inter-onset intervals (IOI) of successive components in tonal sequences. Temporal discrimination was examined using reference stimulus patterns consisting of five 50-ms, 4000-Hz components with equal tonal IOIs selected from the range 100-600 ms. Discrimination was examined in separate conditions by measuring the relative difference limen (DL) for increments of tonal IOI in comparison sequences. In some conditions, comparison sequences featured equal increments of all tonal lOIs to examined listener sensitivity to uniform changes of sequence rate, or tempo. Other conditions measured the DL for increments of a single target IOI within otherwise uniform-rate comparison sequences. For these measurements, the single target IOI was either fixed in sequence location, or randomized in location across listening trials. Listeners in the study included four groups of young and elderly adults with and without high-frequency hearing loss. The results for all listeners showed the relative DL for rate discrimination to decrease from a maximum at the 100-ms IOI to a smaller stable value across the range of longer sequence IOI. All listeners also exhibited larger relative DLs for discrimination of single target intervals compared to rate discrimination for equivalent reference IOI values. Older listeners showed poorer performance than younger listeners in all conditions, with the largest age differences observed for discrimination of brief single intervals that were varied randomly in sequence location. None of the results revealed significant effects of hearing loss on performance of younger and older listeners. PMID- 11425138 TI - The effects of compression ratio, signal-to-noise ratio, and level on speech recognition in normal-hearing listeners. AB - Previous research has demonstrated reduced speech recognition when speech is presented at higher-than-normal levels (e.g., above conversational speech levels), particularly in the presence of speech-shaped background noise. Persons with hearing loss frequently listen to speech-in-noise at these levels through hearing aids, which incorporate multiple-channel, wide dynamic range compression. This study examined the interactive effects of signal-to-noise ratio (SNR), speech presentation level, and compression ratio on consonant recognition in noise. Nine subjects with normal hearing identified CV and VC nonsense syllables in a speech-shaped noise at two SNRs (0 and +6 dB), three presentation levels (65, 80, and 95 dB SPL) and four compression ratios (1:1, 2:1, 4:1, and 6:1). Stimuli were processed through a simulated three-channel, fast-acting, wide dynamic range compression hearing aid. Consonant recognition performance decreased as compression ratio increased and presentation level increased. Interaction effects were noted between SNR and compression ratio, as well as between presentation level and compression ratio. Performance decrements due to increases in compression ratio were larger at the better (+6 dB) SNR and at the lowest (65 dB SPL) presentation level. At higher levels (95 dB SPL), such as those experienced by persons with hearing loss, increasing compression ratio did not significantly affect speech intelligibility. PMID- 11425139 TI - Modeling the motion of the internal tongue from tagged cine-MRI images. AB - A new technique, tagged Cine-Magnetic Resonance Imaging (tMRI), was used to develop a mechanical model that represented local, homogeneous, internal tongue deformation during speech. The goal was to infer muscle activity within the tongue from tissue deformations seen on tMRI. Measurements were made in three sagittal slices (left, middle, right) during production of the syllable /ka/. Each slice was superimposed with a grid of tag lines, and the approximately 40 tag line intersections were tracked at 7 time-phases during the syllable. A local model, similar to a finite element analysis, represented planar stretch and shear between the consonant and vowel at 110 probed locations within the tongue. Principal strains were calculated at these locations and revealed internal compression and extension patterns from which inferences could be drawn about the activities of the Verticalis, Hyoglossus, and Superior Longitudinal muscles, among others. PMID- 11425140 TI - Verifying a vocal tract model with a closed side-branch. AB - In this article an implementation of a vocal tract model and its validation are described. The model uses a transmission line model to calculate pole and zero frequencies for a vocal tract with a closed side-branch such as a sublingual cavity. In the validation study calculated pole and zero frequencies from the model are compared with frequencies estimated using elementary acoustic formulas for a variety of vocal tract configurations. PMID- 11425141 TI - The acoustic bases for gender identification from children's voices. AB - The purpose of this study was to examine the acoustic characteristics of children's speech and voices that account for listeners' ability to identify gender. In Experiment I, vocal recordings and gross physical measurements of 4-, 8-, 12-, and 16-year olds were taken (10 girls and 10 boys per age group). The speech sample consisted of seven nondiphthongal vowels of American English (/ae/ "had," /E/ "head," /i/ "heed," /I/ "hid," /a/ "hod," /inverted v/ "hud," and /u/ "who'd") produced in the carrier phrase, "Say /hVd/ again." Fundamental frequency (f0) and formant frequencies (F1, F2, F3) were measured from these syllables. In Experiment II, 20 adults rated the syllables produced by the children in Experiment I based on a six-point gender rating scale. The results from these experiments indicate (1) vowel formant frequencies differentiate gender for children as young as four years of age, while formant frequencies and f0 differentiate gender after 12 years of age, (2) the relationship between gross measures of physical size and vocal characteristics is apparent for at least 12- and 16-year olds, and (3) listeners can identify gender from the speech and voice of children as young as four years of age, and with respect to young children, listeners appear to base their gender ratings on vowel formant frequencies. The findings are discussed in relation to the development of gender identity and its perceptual representation in speech and voice. PMID- 11425142 TI - Providing low- and mid-frequency speech information to listeners with sensorineural hearing loss. AB - The present study examined the benefits of providing amplified speech to the low- and mid-frequency regions of listeners with various degrees of sensorineural hearing loss. Nonsense syllables were low-pass filtered at various cutoff frequencies and consonant recognition was measured as the bandwidth of the signal was increased. In addition, error patterns were analyzed to determine the types of speech cues that were, or were not, transmitted to the listeners. For speech frequencies of 2800 Hz and below, a positive benefit of amplified speech was observed in every case, although the benefit provided was very often less than that observed in normal-hearing listeners who received the same increase in speech audibility. There was no dependence of this benefit upon the degree of hearing loss. Error patterns suggested that the primary difficulty that hearing impaired individuals have in using amplified speech is due to their poor ability to perceive the place of articulation of consonants, followed by a reduced ability to perceive manner information. PMID- 11425143 TI - Audio-visual enhancement of speech in noise. AB - A key problem for telecommunication or human-machine communication systems concerns speech enhancement in noise. In this domain, a certain number of techniques exist, all of them based on an acoustic-only approach--that is, the processing of the audio corrupted signal using audio information (from the corrupted signal only or additive audio information). In this paper, an audio visual approach to the problem is considered, since it has been demonstrated in several studies that viewing the speaker's face improves message intelligibility, especially in noisy environments. A speech enhancement prototype system that takes advantage of visual inputs is developed. A filtering process approach is proposed that uses enhancement filters estimated with the help of lip shape information. The estimation process is based on linear regression or simple neural networks using a training corpus. A set of experiments assessed by Gaussian classification and perceptual tests demonstrates that it is indeed possible to enhance simple stimuli (vowel-plosive-vowel sequences) embedded in white Gaussian noise. PMID- 11425144 TI - The speed of pitch resolution in a musical context. AB - In five experiments, we investigated the speed of pitch resolution in a musical context. In experiments 1-3, listeners were presented an incomplete scale (doh, re, mi, fa, sol, la, ti) and then a probe tone. Listeners were instructed to make a rapid key-press response to probe tones that were relatively proximal in pitch to the last note of the scale (valid trials), and to ignore other probe tones (invalid trials). Reaction times were slower if the pitch of the probe tone was dissonant with the expected pitch (i.e., the completion of the scale, or doh) or if the probe tone was nondiatonic to the key implied by the scale. In experiments 4 and 5, listeners were presented a two-octave incomplete arpeggio, and then a probe tone. In this case, listeners were asked to make a rapid key-press response to probe tones that were relatively distant in pitch from the last note of the arpeggio. Under these conditions, registral direction and pitch proximity were the dominant influences on reaction time. Results are discussed in view of research on auditory attention and models of musical pitch. PMID- 11425145 TI - In situ source level and source position estimates of biological transient signals produced by snapping shrimp in an underwater environment. AB - Biological transient signals produced by snapping shrimp are sensed underwater by a wide aperture array. The instantaneous range and bearing of the source position of each snap is estimated along with a source level equal to the peak-to-peak amplitude of the pressure impulse generated by the snap at a standard distance of 1 m from its point of origin. For a sample of 1000 snaps recorded in Sydney Harbour, the distribution of peak-to-peak sound pressure levels has a mean value of 187 dB (re 1 microPa) and an interquartile range of 185-189 dB (re 1 microPa). Plotting the Cartesian coordinates of the source positions of the biological transient signals over a period of time maps the two-dimensional spatial distribution of the local snapping shrimp population. The principal habitat is found to be geocoincident with a 120-m-long wharf, the closest point of which is 60 m from the middle of the receiving array. The passive ranging performance of the wide aperture array is evaluated by generating mechanical transient signals at selected positions along the wharf. Precise estimates of the relative times-of arrival of the acoustic wavefronts lead to source range and bearing estimates with standard deviations of only 0.1 m and 0.005 degrees (respectively), in agreement with theoretical predictions. PMID- 11425146 TI - Localization and visual verification of a complex minke whale vocalization. AB - A recently described population of minke whales (Balaenoptera acutorostrata) offered a unique opportunity to study its acoustic behavior. The often inquisitive dwarf minke whale is seen on the Great Barrier Reef nearly coincident with its suspected calving and breeding seasons. During drifting encounters with whales, a towed hydrophone array was used to record sounds for subsequent localization of sound sources. Shipboard and in-water observers linked these sounds to the closely circling minke whale. A complex and stereotyped sound sequence, the "star-wars" (SW) vocalization, was recorded during a series of visual and acoustic observations. The SW vocalization spanned a wide frequency range (50 Hz-9.4 kHz) and was composed of distinct and stereotypically repeated units with both amplitude and frequency-modulated components. Broadband source levels between 150 and 165 dB re 1 microPa at 1 m were calculated. Passive acoustic studies can utilize this distinct vocalization to help determine the behavior, distribution, and movements of this animal. While the SW vocalization's function remains unknown, the regularly repeated and complex sound sequence was common in low latitude, winter month aggregations of minke whales. At this early stage, the SW vocalization appears similar to the songs of other whale species and has characteristics consistent with those of reproductive advertisement displays. PMID- 11425147 TI - Ultrasound detection by clupeiform fishes. AB - It has previously been shown that at least one species of fish (the American shad) in the order clupeiforms (herrings, shads, and relatives) is able to detect sounds up to 180 kHz. However, it has not been clear whether other members of this order are also able to detect ultrasound. It is now demonstrated, using auditory brainstem response (ABR), that at least one additional species, the gulf menhaden (Brevoortia patronus), is able to detect ultrasound, while several other species including the bay anchovy (Anchoa mitchilli), scaled sardine (Harengula jaguana), and Spanish sardine (Sardinella aurita) only detect sounds to about 4 kHz. ABR is used to confirm ultrasonic hearing in the American shad. The results suggest that ultrasound detection may be limited to one subfamily of clupeiforms, the Alosinae. It is suggested that ultrasound detection involves the utricle of the inner ear and speculate as to why, despite having similar ear structures, only one group may detect ultrasound. PMID- 11425148 TI - Ultrasound phased arrays for prostate treatment. AB - The effect of array geometry on the steering performance of ultrasound phased arrays is examined theoretically, in order to maximize array performance under the given anatomical constraints. This paper evaluates the performance of arrays with spherical and cylindrical geometry, determined by using computer simulations of the pressure fields produced at various extremes of steering. The spherical segment arrays were truncated for insertion into the rectum, and contained either annular or linear elements. The cylindrical arrays were either flat or had a variable curvature applied along their length. Fields were computed by dividing the array elements into many point sources. The effectiveness of an array configuration when steered to a particular focal location was assessed by defining a parameter, G, as the ratio of the intensity at the desired focus to the maximum intensity of any unwanted lobes. The performance of truncated spherical arrays with annular elements was evaluated for focal steering along the array axis (in depth, in the z direction). When steered 15 mm toward the source, these truncated spherical annular arrays exhibited excellent performance, with G>5.7 for arrays containing more than 10 elements. Similarly, the spherical arrays with linear elements performed well when steered along the array axis to the same degree, with G>7 (for element widths up to 3 lambda), though many more array elements were required. However, when these arrays were steered 15 mm laterally, along the length of the prostate (the y direction), the value for G fell below 1 for element widths greater than about 1.6 lambda. It was found that the cylindrical arrays performed much better for y-direction steering (G>4, for 60 mm arrays with an element width of 1.75 lambda), but their performance was poorer when steered in the z direction (G approximately 4 for an element width of 1.5 lambda). In order to find a compromise between these extremes, a curved cylindrical array was examined, which was a cylindrical array with additional curvature along its length. These curved cylindrical arrays yielded performance between that of spherical linear arrays and cylindrical arrays, with better steering along the y direction than the spherical arrays and better z-direction steering than the cylindrical arrays. PMID- 11425152 TI - Hyperbolic location errors due to insufficient numbers of receivers. PMID- 11425153 TI - Ants are deaf. PMID- 11425154 TI - CR1 density polymorphism on erythrocytes of falciparum malaria patients in Thailand. AB - Complement receptor type 1 (CR1) on erythrocytes shows an inherited numerical polymorphism which correlates with a HindIII-RFLP (restriction fragment length polymorphism) of the CR1 gene in various populations. To investigate the relationship between CR1 density polymorphism and disease severity, we typed 185 Thai patients with acute falciparum malaria (55 severe and 130 uncomplicated) for their genotypes of this polymorphism. The level of expression of erythrocyte CR1 from 42 randomly selected patients was measured by enzyme-linked immunosorbent assay (ELISA). We observed a significantly higher frequency of homozygotes of the CR1 low density allele (LL) among the severe group as compared to the uncomplicated group (P = 0.005). CR1 expression on erythrocytes from patients with the LL genotype was significantly lower than homozygotes with the high density allele (HH) (P < 0.0001) and heterozygotes (HL) (P = 0.013). The results suggest that a genetically-determined low CR1 density on erythrocytes may be a risk factor for developing a more severe form of malaria in Thai subjects. PMID- 11425155 TI - Multiple genotypes of St. Louis encephalitis virus (Flaviviridae: Flavivirus) circulate in Harris County, Texas. AB - St. Louis encephalitis (SLE) is endemic in Harris County, Texas. The disease is a public health concern in Houston, the largest city in Harris County, and in the state. Consequently, intensive surveillance for SLE virus in local mosquito populations is carried out by the Harris County Mosquito Control Division each year. In this study, we examined genetic variation among SLE isolates obtained during routine virus surveillance over a 13-year time period (1986-1999). St. Louis encephalitis virus isolates were tested for genetic variation using reverse transcription-polymerase chain reaction followed by single-strand conformation polymorphism (SSCP). The results indicated that multiple genotypes of the virus circulate in Harris County. During several years, the genotypes were restricted in their location, i.e., each general area within the county had a specific genotype of the virus. In other years, the various genotypes were widely distributed throughout the county. The presence of multiple distinct genotypes suggests that viruses with different biological characteristics may be circulating in Harris County, and that discrete foci of SLE virus activity occur simultaneously. PMID- 11425156 TI - Diagnosis of imported malaria by Plasmodium lactate dehydrogenase (pLDH) and histidine-rich protein 2 (PfHRP-2)-based immunocapture assays. AB - This study was conducted to evaluate the performance of two rapid non-microscopic assays: Plasmodium lactate dehydrogenase (pLDH) assay (OptiMAL) and Plasmodium falciparum histidine-rich protein 2 (PfHRP-2) assay (ICT Malaria). The assays were used to detect malaria infection in 515 immigrants living in Kuwait. The performance of both assays was compared to that of microscopy of Giemsa-stained thick blood films and to each other. Of the 515 patients tested, 163 were positive for malaria parasites by microscopy of thick blood film. Of these, 87 were infected with Plasmodium vivax parasites, 63 with P. falciparum, 1 with Plasmodium malariae, and 12 had mixed infections of P. falciparum and P. vivax. The PfHRP-2 assay detected 53 P. falciparum infections and, as expected, failed to detect all but one case of P. vivax. Three cases of mixed infections were also not detected by this assay. The pLDH assay detected 56 P. falciparum cases and 77 P. vivax infections but failed to detect 4 cases of mixed infections. Compared to microscopy, the performance of both the assays to diagnose P. falciparum infection was comparable. The sensitivity for the PfHRP-2 assay was 82% with a specificity of 99.0% and for the pLDH assay the sensitivity was 89% with a specificity of 99.5%. The PfHRP-2 assay detected 4 false positive cases, 2 of which were also detected by the pLDH assay. These patients reported treatment with chloroquine in the last 2-5 weeks. Though the immunocapture diagnostic assays may be helpful in certain situations, microscopy of thick blood film is still the method of choice in diagnosing imported malaria. PMID- 11425157 TI - Maintenance therapy of melioidosis with ciprofloxacin plus azithromycin compared with cotrimoxazole plus doxycycline. AB - This is a report of a randomized, open, labeled study of the maintenance treatment of melioidosis using a combination of ciprofloxacin and azithromycin (Regimen A) for 12 weeks versus a combination of cotrimoxazole and doxycycline (Regimen B) for 20 weeks. The study was conducted at two tertiary-care hospitals in northeast Thailand. A total 65 patients were enrolled, 36 and 29, respectively, between August 1997 and July 1998. Subjects were randomly allocated to each arm of the trial, resulting in 32 treated under Regimen A and 33 in B. The main outcome was a culture-proven relapse in melioidosis. There were more relapses under Regimen A at 22% (7 of 32) than in Regimen B, 3% (1 of 33). The 19% difference in the rates was significant (95% confidence interval [CI]: 3% to 34%; exact P-value = 0.027). Based on our data, a combination of cotrimoxazole and doxycycline treatment for 20 weeks should be given further consideration as the maintenance therapy of choice for melioidosis. PMID- 11425158 TI - Short report: differences in dihydrofolate reductase but not dihydropteroate synthase alleles in Plasmodium falciparum isolates from geographically distinct areas in Malaysia. AB - Dihydropteroate synthase (dhps) and dihydrofolate reductase (dhfr) alleles were typed in 67 Malaysian Plasmodium falciparum isolates. The isolates were collected from two geographically distinct locations: 51 from Sabah, Malaysian Borneo, where sulfadoxine/pyrimethamine (SDX/PYR) is used to treat uncomplicated malaria and 16 from Peninsular Malaysia where in vivo resistance to SDX/PYR has been reported. A total of seven dhps alleles were identified with no significant difference in allele frequency between the 2 populations. Two of the dhps alleles described here have not been previously reported. Four dhfr alleles were detected in 67 P. falciparum isolates. Eighty-seven percent of the isolates from the Peninsula, where clinical SDX/PYR failure has been reported, had dhfr alleles with triple point mutations while all of the isolates from Sabah had dhfr alleles with 2 or less point mutations. The difference in dhfr allele frequency between the two populations was highly significant. There was no correlation between in vitro PYR response and accumulation of dhfr point mutations. PMID- 11425159 TI - Comparison of direct and membrane feeding methods to infect Anopheles arabiensis with Plasmodium falciparum. AB - Two standard methods are available to infect mosquitoes with malaria parasites: direct feeding through the skin of the gametocyte carrier, and membrane feeding. Anopheles arabiensis collected at larval stages and reared in an insectary were fed in parallel by feeding on Plasmodium falciparum gametocyte carriers and by membrane feeding on venous blood of the same gametocyte carriers. Infection of mosquitoes was assessed at Day 7 post bloodmeal by oocyst count of the mosquito midguts. The following parameters were not significantly different between the two methods: the percentage of gametocyte carriers infective for at least one mosquito (52.4% through the skin versus 57.1% through the membrane), the mean infection rate of mosquitoes (10.0% versus 11.3%), the geometric mean oocyst number per mosquito (2.51 versus 3.83). In conclusion, infection of mosquitoes by membrane feeding was similar to infection by direct feeding. Most of the volunteers preferred venipuncture to mosquito bites. PMID- 11425160 TI - Interleukin levels in cerebrospinal fluid from children with neurocysticercosis. AB - No information about the levels of pro-inflammatory interleukins has been described in children with neurocysticercosis (NCC). The levels of tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-5, IL-6, and IL-12 in the cerebrospinal fluid from children with NCC were determined by enzyme-linked immunosorbent assay (ELISA). Twelve children with NCC, six with active and six with inactive disease, and six children without NCC were studied. TNF-alpha was undetectable in CSF from controls and five children with inactive NCC, whereas the levels were significantly higher (median 22.1 pg/ml; P = 0.008) in all children with active NCC. Levels of IL-6 were low in active and inactive NCC patients but two subjects with active subarachnoid disease had high levels. IL-5 and IL-12 were not detected. This study shows that high levels of TNF-alpha are present in CSF from children with active NCC. IL-6 levels are higher when infection occurs in the subarachnoid space. PMID- 11425161 TI - T cell receptor Vbeta gene usage in Thai children with dengue virus infection. AB - T lymphocyte activation during dengue is thought to contribute to the pathogenesis of dengue hemorrhagic fever (DHF). We examined the T cell receptor Vbeta gene usage by a reverse transcriptase-polymerase chain reaction assay during infection and after recovery in 13 children with DHF and 13 children with dengue fever (DF). There was no deletion of specific Vbeta gene families. We detected significant expansions in usage of single Vbeta families in six subjects with DHF and three subjects with DF over the course of infection, but these did not show an association with clinical diagnosis, viral serotype, or HLA alleles. Differences in Vbeta gene usage between subjects with DHF and subjects with DF were of borderline significance. These data suggest that the differences in T cell activation in DHF and DF are quantitative rather than qualitative and that T cells are activated by conventional antigen(s) and not a viral superantigen. PMID- 11425162 TI - Comparative neurovirulence of attenuated and non-attenuated strains of Venezuelan equine encephalitis virus in mice. AB - A candidate live-attenuated virus vaccine for protection against Venezuelan equine encephalitis (VEE) (designated V3526) was tested in mice to measure the magnitude, duration, and kinetics of virus replication in the blood and the central nervous system and its phenotypic stability after multiple passages in mice and cell culture. All results were compared to parallel experiments with parental virus and the existing VEE virus vaccine, TC-83. Maximum virus titers in the brains of V3526-inoculated mice were between 10- and 100-fold less than those observed in brains of mice inoculated intracranially (i.c.) with either the parental virus or TC-83. Neither V3526 nor TC-83 was lethal in BALB/c mice inoculated i.c.. However, mice inoculated with TC-83 developed acute symptoms lasting at least 14 days. In contrast, i.c. inoculation of TC-83 was uniformly lethal for C3H/HeN mice. V3526 was avirulent in both BALB/c and C3H/HeN mice after i.c. inoculation. The virulence characteristics of V3526 remained unchanged after five serial i.c. passages in mouse brains or after five cell culture passages. Finally, pathologic changes induced after i.c. inoculation of V3526 were consistently less severe and of shorter duration than those observed in TC 83-inoculated mice. Based on these results, V3526 is stable and appears to be significantly less neurovirulent in mice than TC-83. PMID- 11425163 TI - Short report: documentation of iodine deficiency in Haitian schoolchildren: implication for lymphatic filariasis elimination in Haiti. AB - In this study we documented unexpected moderate-to-severe iodine deficiency in Haitian schoolchildren although they live in a coastal community where presumably they have access to iodine-containing seafood. This fact combined with the lack of an iodized salt supply and endemic lymphatic filariasis makes community distribution of diethylcarbamazine-fortified, iodized salt an attractive strategy for elimination of lymphatic filariasis and iodine deficiency disorders in this area of Haiti. Combining lymphatic filariasis elimination with other public health interventions is one strategy to increase its public health benefit and maximize the impact of limited public health resources. PMID- 11425164 TI - Hemoglobin concentration in children in a malaria holoendemic area is determined by cumulated Plasmodium falciparum parasite densities. AB - In malaria holoendemic areas children are anemic, but the exact influence of falciparum malaria on hemoglobin (Hb) concentration remains largely unsettled. Prospective data were therefore collected in children < 24 months of age during five months in a Tanzanian village. Children with mean asymptomatic parasitemia > or = 400/microl had lower median Hb levels during the study than those with mean density < 400/microl. The difference was 9.7 g/L (95% confidence interval [CI] 2.8-17). In children with one or more clinical malaria episodes, the median Hb was 8.3 g/L (95% CI 0.9-16) lower than those without episode. If early treatment failure was recorded, the immediate effect on Hb was particularly important with a mean drop of 17 g/L. Interestingly, at study-end the Hb concentration represented a function of the area under the parasitemia curve (AUPC) during the previous five months, adjusting for age. In conclusion, stepwise deterioration in median Hb levels was found by asymptomatic parasitemia, clinical malaria episode, and most significantly, treatment failure. PMID- 11425165 TI - Short report: increased susceptibility to Plasmodium malariae in pregnant alpha(+)-thalassemic women. AB - The influence of alpha(+)-thalassemia on malaria in pregnancy was assessed in a cross-sectional study of 530 women in Ghana. Plasmodial infections, alpha(+) thalassemia, serum levels of C-reactive protein, and antimalarial drugs in urine were determined. The alpha-globin genotypes did not correlate with the prevalence of Plasmodium falciparum-infection and parasite densities. However, Plasmodium malariae tended to be more frequent in alpha(+)-thalassemic women (P = 0.05). Excluding women with residual antimalarials, a significant excess of P. malariae was observed in alpha(+)-thalassemic individuals. Febrile responses (P = 0.05) and inflammation (CRP > 0.6 mg/dl, P = 0.06) appeared to be less common in infected alpha(+)-thalassemic women and were also comparatively rare in parasitemic individuals who harbored double species infections with P. falciparum and P. malariae. Plasmodium malariae may influence the pathogenesis of falciparum malaria leading to a low prevalence of inflammation and febrile responses in alpha(+)-thalassemic women. PMID- 11425166 TI - The dengue and dengue hemorrhagic fever epidemic in Puerto Rico, 1994-1995. AB - From June 1, 1994 to May 31, 1995 a total of 24,700 cases of dengue (7.01/1,000 population) were reported to the laboratory-based surveillance system in Puerto Rico (1991-1994, annual average: 2.55/1,000). Dengue virus 2 predominated. The earliest indicator of epidemic activity was the virus isolation rate in May 1994 (14.0% versus 5.7% average). The male-to-female ratio among cases was 1:1.1; 65.4% were younger than 30 years (the 10 to 19 year age group had the highest incidence, 11.8/1,000). At least 5,687 cases (23.0%) showed a hemorrhagic manifestation; 4,662 (18.9%) were hospitalized, and 40 died (0.2%; 10 laboratory positive). Two cases documented by laboratory were transmitted by unusual routes- intrapartum and through a bone marrow transplant. Among 2,004 hospitalized cases reported by infection control nurses, 139 (6.9%) fulfilled the criteria for dengue hemorrhagic fever (DHF) and another 13 cases (0.6%) had dengue shock syndrome. This epidemic produced the largest number of hospitalizations, DHF cases, and deaths from any dengue epidemic in Puerto Rico. Severity did not change throughout the year. Surveillance capabilities were maintained by temporary, simplified reporting methods, none of which could be recommended as the single method of choice for surveillance; each must be used (on site, or as a service available from a reference laboratory) at the right time in the epidemic cycle. The utility of comparisons of current and previous data underscores the value of long-term surveillance. Our analysis was unable to document whether significantly increased transmission occurred more often in cities where the water supply was rationed or where the local landfill was closed. PMID- 11425167 TI - Dengue activity in Puerto Rico during an interepidemic period (1995-1997). AB - From 1995 to 1997 dengue was reported in Puerto Rico at an average annual rate of 1.75/1,000 population, compared to 6.73 in 1994, an epidemic year. Dengue virus serotypes 1 (DEN-1), -2, and -4 were isolated each year, with DEN-2 predominating in 1995 and 1996, and DEN-4 in 1997. From 1995 through 1997 incidence was highest (0.61-0.77/1,000) in persons under 30 years of age; males and females were equally affected. Among positive cases, 28.3% to 37.9% were hospitalized; 28.9% to 35.2% had hemorrhagic manifestations; at least 1.1% to 1.6% fulfilled the criteria for dengue hemorrhagic fever/dengue shock syndrome; and 0.2% to 0.3% died. Neither hurricane preparations (1995) nor widespread floods (1996) seem to have affected dengue incidence. Most municipalities with the highest laboratory diagnosed dengue rates in 1995 were in the eastern foothills of the central mountains, an area relatively spared by the 1994 epidemic. In the next two years, at least half of the municipalities with the highest laboratory-diagnosed dengue rates were in the west. The most intense municipal outbreak of this period (DEN 2, Villalba, 1995, rate of 11.67/1,000) is described to highlight the importance of local conditions and epidemiologic history in determining the risk of dengue. PMID- 11425168 TI - Ecological studies of enzootic Venezuelan equine encephalitis in north-central Venezuela, 1997-1998. AB - From 1997-1998, we investigated the possible continuous circulation of epizootic Venezuelan equine encephalitis (VEE) virus suggested by a 1983 subtype IC interepizootic mosquito isolate made in Panaquire, Miranda State, Venezuela. The study area was originally covered by lowland tropical rainforest but has been converted into cacao plantations. Sentinel hamsters, small mammal trapping, mosquito collections, and human serosurveys were used to detect active or recent virus circulation. Six strains of subtype ID VEE virus were isolated from hamsters that displayed no apparent disease. Four other arboviruses belonging to group A (Togaviridae: Alphavirus), two Bunyamwera group (Bunyaviridae), and three Gamboa group (Bunyaviridae) arboviruses were also isolated from hamsters, as well as 8 unidentified viruses. Venezuelan equine encephalitis-specific antibodies were detected in 5 small mammal species: Proechimys guairae, Marmosa spp., and Didelphis marsupialis. Mosquito collections comprised of 38 different species, including 8 members of the subgenus Culex (Melanoconion), did not yield any virus isolates. Sera from 195 humans, either workers in the cacao plantation or nearby residents, were all negative for VEE virus antibodies. Sequences of 1,677 nucleotides from the P62 gene of 2 virus isolates indicated that they represent a subtype ID lineage that is distinct from all others characterized previously, and are unrelated to epizootic VEE emergence. PMID- 11425169 TI - Limited polymorphism in Plasmodium falciparum sexual-stage antigens. AB - In areas highly endemic for malaria, individuals are frequently found to be infected simultaneously with multiple Plasmodium falciparum clones. This raises the question of whether all parasite clones produce gametocytes equally or whether gametocytogenesis is suppressed in some clones. In order to assess this in epidemiological studies, polymorphic genes specifically expressed in gametocytes could be analyzed by both amplification of genomic DNA from blood samples and by reverse transcribed polymerase chain reaction amplifying expressed gametocyte-specific genes only. Here we report the analysis of diversity in the three gametocyte-specific genes Pfs16, Pfs48/45, and Pfs230. In addition to the previously published data, limited polymorphism was found in the coding sequences of Pfs16 and Pfs48/45. Larger polymorphism was identified in Pfs230, which might allow the development of a discriminating PCR-based genotyping scheme for transmission studies. However, the limited polymorphism in Pfs16 and Pfs48/45 renders these molecules poorly useful for such studies. PMID- 11425170 TI - Trocara virus: a newly recognized Alphavirus (Togaviridae) isolated from mosquitoes in the Amazon Basin. AB - This report describes Trocara virus, a newly recognized member of the genus Alphavirus, that has been isolated from Aedes serratus mosquitoes collected at two widely separated sites in the Amazon Basin. Biological, antigenic and genetic characteristics of the new virus are given. Results of these studies indicate that Trocara virus is the first member of a newly discovered antigenic complex within the family Togaviridae genus Alphavirus. The public health and veterinary importance of Trocara virus is still unknown. PMID- 11425171 TI - A protocol for geographically randomized snail surveys in schistosomiasis fieldwork using the global positioning system. AB - A protocol was created for performing geographically randomized snail surveys for schistosomiasis research using the global positioning system (GPS). This protocol differs from traditional surveys in its ability to accurately map and measure the spatial distribution of snail habitat. The protocol was used to map irrigation ditches, the primary habitat for Oncomelania hupensis, in two residence areas in Sichuan Province, China. From the 7,450 meters of mapped ditches, snail surveys were performed at 203 random sites along the ditch network. Of these, 116 (57.1%) sites had snails. The total number of living snails captured was 2,014, resulting in an average snail density of 0.27 snails per linear meter of potential habitat. PMID- 11425172 TI - The ears of the hippopotamus: manifestations, determinants, and estimates of the malaria burden. AB - Malarious patients experience asymptomatic parasitemia; acute febrile illness (with cerebral damage, anemia, respiratory distress, hypoglycemia); chronic debilitation (anemia, malnutrition, nervous system-related sequelae); and complications of pregnancy (anemia, low birth weight, increased infant mortality). These manifestations in patients, communities, and countries reflect intrinsic (human, parasite, mosquito) and extrinsic (environmental, social, behavioral, political, and economic conditions as well as disease-control efforts) determinants. At a minimum, between 700,000 and 2.7 million persons die yearly from malaria, over 75% of them African children. Between 400 and 900 million acute febrile episodes occur yearly in African children under 5 yr of age living in endemic areas. Although about half of these children are parasitemic, all merit consideration of malaria-specific therapy, which is becoming more problematic because of parasite resistance to drugs. These numbers will more than double over the next 20 yr without effective control. Fewer than 20% of these febrile episodes and deaths come to the attention of any formal health system. The relatively few ill patients who have any contact with the health services represent the "ears of the hippopotamus." Greatly intensified research activities and control of the intolerable burden of malaria are mandatory if economic development is to accelerate in Africa. In particular, support should be targeted to understanding and preventing malaria-induced anemia, hypoglycemia, effects on pregnancy, and neurologic and developmental impairment. To decrease and stop transmission of this intolerable scourge, there is an urgent need for malaria vaccines, newer drugs, and better vector control methods as well as the ability to improve current technologies and use them more efficiently. PMID- 11425173 TI - The public health impact of chloroquine resistance in Africa. AB - Between 1978 and 1988 Plasmodium falciparum resistance to chloroquine has been reported in all countries of tropical Africa. Despite the intensification of resistance during the last 2 decades, chloroquine remains in 2000 the first-line treatment for malaria in most of these countries. Here we review published data on the public health impact of antimalarial drug resistance in Africa. These data show that since the late 1980s convincing evidence of a major public health impact of the spread of chloroquine resistance has been available. Hospital studies in various African countries have documented a 2- or 3-fold increase in malaria deaths and admissions for severe malaria, an increase temporally related to the emergence of chloroquine resistance. Data from sentinel demographic surveillance systems in Senegal indicated that mortality attributable to malaria in children increased by as much as 6-fold among populations where low levels of malaria mortality had been achieved because of efficient health services before the emergence of chloroquine resistance. Increasing incidence of severe malarial anemia also contributed to human immunodeficiency virus dissemination. The dramatic impact of chloroquine resistance on malaria mortality has long been underestimated because only a low proportion of malaria attacks are potentially lethal among persons continuously exposed since birth to high levels of transmission. There is an urgent need to change treatment policies in Africa. PMID- 11425174 TI - All-cause mortality among young children in western Kenya. VI: the Asembo Bay Cohort Project. AB - Although all-cause mortality has been used as an indicator of the health status of childhood populations, such data are sparse for most rural areas of sub Saharan Africa, particularly community-based estimates of infant mortality rates. The longitudinal follow-up of more than 1,500 children enrolled at birth into the Asembo Bay Cohort Project (ABCP) in western Kenya between 1992 and 1996 has provided a fixed birth cohort for estimating all-cause mortality over the first 5 yr of life. We surveyed mothers and guardians of cohort children in early 1999 to determine survival status. A total of 1,260 households were surveyed to determine the survival status of 1,556 live births (99.2% of original cohort, n = 1,570). Most mothers (66%) still resided but 27.5% had migrated, and 5.5% had died. In early 1999, the overall cumulative incidence of all-cause mortality for the entire 1992-1996 birth cohort was 26.5% (95% confidence interval, 24.1-28.9%). Neonatal and infant mortality were 32 and 176 per 1,000 live births, respectively. These community-based estimates of mortality in the ABCP area are substantially higher than for Kenya overall (nationally, infant mortality is 75 per 1,000 live births). The results provide a baseline description of all-cause mortality among children in an area with intense Plasmodium falciparum transmission and will be useful in future efforts to monitor changes in death rates attributable to control programs for specific diseases (e.g., malaria and HIV/AIDS) in Africa. PMID- 11425175 TI - The burden of malaria in pregnancy in malaria-endemic areas. AB - Pregnant women in malarious areas may experience a variety of adverse consequences from malaria infection including maternal anemia, placental accumulation of parasites, low birth weight (LBW) from prematurity and intrauterine growth retardation (IUGR), fetal parasite exposure and congenital infection, and infant mortality (IM) linked to preterm-LBW and IUGR-LBW. We reviewed studies between 1985 and 2000 and summarized the malaria population attributable risk (PAR) that accounts for both the prevalence of the risk factors in the population and the magnitude of the associated risk for anemia, LBW, and IM. Consequences from anemia and human immunodeficiency virus infection in these studies were also considered. Population attributable risks were substantial: malaria was associated with anemia (PAR range = 3-15%), LBW (8-14%), preterm-LBW (8-36%), IUGR-LBW (13-70%), and IM (3-8%). Human immunodeficiency virus was associated with anemia (PAR range = 12-14%), LBW (11-38%), and direct transmission in 20-40% of newborns, with direct mortality consequences. Maternal anemia was associated with LBW (PAR range = 7-18%), and fetal anemia was associated with increased IM (PAR not available). We estimate that each year 75,000 to 200,000 infant deaths are associated with malaria infection in pregnancy. The failure to apply known effective antimalarial interventions through antenatal programs continues to contribute substantially to infant deaths globally. PMID- 11425176 TI - The epidemiology and burden of Plasmodium falciparum-related anemia among pregnant women in sub-Saharan Africa. AB - The paucity of precise information on the burden of malaria among pregnant women has hampered effective lobbying for the inclusion of preventative strategies against malaria in Safe Motherhood Initiatives. This article reviews the evidence on the coincidental risks of malaria and anemia in Africa and attempts to estimate the probable burden of malaria-related severe anemia in this susceptible group. Twenty-six studies on hemoglobin levels in all-parity pregnant women throughout this region could be matched with a malaria parasite ratio in children < 15 yr old (a measure of the intensity of transmission). In areas with no malaria, the mean hemoglobin levels were markedly higher than those found in areas with stable malaria transmission, though changes with increasing intensity of transmission were unclear. Eighteen studies from areas with stable malaria transmission in sub-Saharan Africa suggested that the median prevalence of severe anemia in all-parity pregnant women is approximately 8.2%. Assuming that 26% of these cases are due to malaria, it is suggested that as many as 400,000 pregnant women may have developed severe anemia as a result of infection with malaria in sub-Saharan Africa in 1995. PMID- 11425177 TI - The cost-effectiveness of antenatal malaria prevention in sub-Saharan Africa. AB - Antimalarial chemoprophylaxis during pregnancy significantly increases the birth weight of babies born to primigravidae, but coverage in sub-Saharan Africa is very limited. This analysis assessed whether increasing coverage is justified on cost-effectiveness grounds. A standardized modeling framework was used to estimate ranges for the cost per discounted year of life lost averted by weekly chloroquine chemoprophylaxis and intermittent sulfadoxine-pyrimethamine (SP) treatment for primigravidae in an operational setting with moderate to high malaria transmission. The SP regimen was found to be more cost-effective than the chloroquine regimen, because of both lower costs and higher compliance. Both regimens appear to be a good value for money in comparison with other methods of malaria control and based on rough cost-effectiveness guidelines for low-income countries, even with high levels of drug resistance. However, extending the SP regimen to all gravidae and increasing the number of doses per pregnancy could make the intervention significantly less cost-effective. PMID- 11425178 TI - Gaps in the childhood malaria burden in Africa: cerebral malaria, neurological sequelae, anemia, respiratory distress, hypoglycemia, and complications of pregnancy. AB - Evaluations of the African childhood malaria burden do not fully quantify the contributions of cerebral malaria (CM), CM-associated neurological sequelae, malarial anemia, respiratory distress, hypoglycemia, and pregnancy-related complications. We estimated the impact of these malaria manifestations on members of the African population < 5 years old. Calculations were based on an extensive literature review that used National Library of Medicine search engines, other bibliographic sources, and demographic data. In sub-Saharan Africa, CM annually affects 575,000 children < 5 years of age and 110,000 (approximately 19% case fatality rate [CFR]) die. Childhood survivor, of CM experience developmental and behavioral impairments: each year, 9,000-19,000 children (> 2% of survivors of CM) < 5 years of age in Africa experience neurological complications lasting > 6 months. Severe malarial anemia heavily burdens hospitals with rising admission and CFRs and with treatments that are complicated by limited and sometimes contaminated blood supplies. Severe malarial anemia occurs 1.42-5.66 million times annually and kills 190,000-974,000 (> 13% CFR) children < 5 years of age annually. Respiratory distress, hypoglycemia, and overlapping clinical manifestations cause 1.12-1.99 million cases and > 225,000 (> 18% CFR) additional deaths among African children with malaria. Maternal, placental, or fetal malaria infection during pregnancy adversely affects development and survival of fetuses and newborns through low birth weight (LBW), maternal anemia, and possibly abortion and stillbirth. Between 167,000 and 967,000 cases of malaria-associated LBW occur yearly; malaria-induced LBW kills 62,000-363,000 (> 38% CFR) newborns each year. All the gaps in the burden comprise 0.4-1.7 million deaths annually, > 50% of which are due to severe malarial anemia. These malaria-induced medical problems constitute major clinical, public health, and research challenges in that they may contribute to more than double the mortality than is generally acknowledged. PMID- 11425179 TI - Impact of Plasmodium falciparum malaria on performance and learning: review of the evidence. AB - Despite the growing recognition that Plasmodium falciparum malaria constitutes a major threat to child survival, the indirect consequences of disease and infection on general human development have been less well described. This review suggests that malaria in childhood is likely to have effects on general cognitive and behavioral development, which range from subtle to profound. Nevertheless, our understanding of the numbers of affected children, and the persistence of and recovery from impairment remains ill defined. Only through large long-term studies will we be able to establish the wider consequences of malaria on communities in areas of the world where malaria is endemic. PMID- 11425180 TI - Toward a framework and indicators for monitoring Roll Back Malaria. AB - Roll Back Malaria (RBM) is a new global partnership that aims to halve the malaria burden by the year 2010. A framework and indicators for monitoring the outcomes and impact of RBM have been defined through an extensive consultative process. The framework identifies critical areas for monitoring RBM action relating to 1) the impact on malaria burden, 2) improvements in malaria prevention and treatment, 3) related health sector development, and 4) support for RBM action (including partnerships). A set of RBM indicators has been defined that corresponds to these critical areas but that reflects the major variations in malaria epidemiology and the principal interventions in different parts of the world. Countries would select indicators that are appropriate for their situation. Four global indicators are proposed for use by all countries in which RBM action is under way. Data collection procedures are discussed, and it is indicated how monitoring RBM action can build on existing data-collection mechanisms. PMID- 11425181 TI - The economic burden of malaria. AB - Malaria and poverty are intimately connected. Controlling for factors such as tropical location, colonial history, and geographical isolation, countries with intensive malaria had income levels in 1995 of only 33% that of countries without malaria, whether or not the countries were in Africa. The high levels of malaria in poor countries are not mainly a consequence of poverty. Malaria is geographically specific. The ecological conditions that support the more efficient malaria mosquito vectors primarily determine the distribution and intensity of the disease. Intensive efforts to eliminate malaria in the most severely affected tropical countries have been largely ineffective. Countries that have eliminated malaria in the past half century have all been either subtropical or islands. These countries' economic growth in the 5 years after eliminating malaria has usually been substantially higher than growth in the neighboring countries. Cross-country regressions for the 1965-1990 period confirm the relationship between malaria and economic growth. Taking into account initial poverty, economic policy, tropical location, and life expectancy, among other factors, countries with intensive malaria grew 1.3% less per person per year, and a 10% reduction in malaria was associated with 0.3% higher growth. Controlling for many other tropical diseases does not change the correlation of malaria with economic growth, and these diseases are not themselves significantly negatively correlated with economic growth. A second independent measure of malaria has a slightly higher correlation with economic growth in the 1980-1996 period. We speculate about the mechanisms that could cause malaria to have such a large impact on the economy, such as foreign investment and economic networks within the country. PMID- 11425183 TI - The malaria burden and Africa. PMID- 11425182 TI - The neglected burden of Plasmodium vivax malaria. AB - We estimate that the global burden of malaria due to Plasmodium vivax is approximately 70-80 million cases annually. Probably approximately 10-20% of the world's cases of P. vivax infection occur in Africa, south of the Sahara. In eastern and southern Africa, P. vivax represents around 10% of malaria cases but < 1% of cases in western and central Africa. Outside of African, P. vivax accounts for > 50% of all malaria cases. About 80-90% of P. vivax outside of Africa occurs in the Middle East, Asia, and the Western Pacific, mainly in the most tropical regions, and 10-15% in Central and South America. Because malaria transmission rates are low in most regions where P. vivax is prevalent, the human populations affected achieve little immunity to this parasite; as a result, in these regions, P. vivax infections affect people of all ages. Although the effects of repeated attacks of P. vivax through childhood and adult life are only rarely directly lethal, they can have major deleterious effects on personal well being, growth, and development, and on the economic performance at the individual, family, community, and national levels. Features of the transmission biology of P. vivax give this species greater resilience than the less robust Plasmodiumfalciparum in the face of conditions adverse to the transmission of the parasites. Therefore, as control measures become more effective, the residual malaria burden is likely increasingly to become that of P. vivax. PMID- 11425184 TI - The malaria burden and the need for research and capacity strengthening in Africa. PMID- 11425185 TI - The intolerable burden of malaria: a new look at the numbers. PMID- 11425187 TI - Conjugation of yeast mannans with protein employing cyanopyridinium agent (CDAP)- an effective route of antifungal vaccine preparation. AB - The possibility of using 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) for activation of saccharide hydroxyl groups (instead of hazardous cyanogen bromide) is examined with cell-surface mannans of the yeasts Candida albicans, Candida tropicalis, Candida lambica and galactoglucoxylomannan of Cryptococcus laurentii. Direct conjugation with human serum albumin yielded soluble products with increased molecular size in comparison with the original polysaccharides. Immunodiffusion experiments revealed that conjugation did not affect the immunospecificity of the antigen epitppe. PMID- 11425186 TI - The sialyl-alpha2,6-lactosaminyl-structure: biosynthesis and functional role. AB - Sialylation represents one of the most frequently occurring terminations of the oligosaccharide chains of glycoproteins and glycolipids. Sialic acid is commonly found alpha2,3- or alpha2,6-linked to galactose (Gal), alpha2,6-linked to N acetylgalactosamine (GalNAc) or alpha2,8-linked to another sialic acid. The biosynthesis of the various linkages is mediated by the different members of the sialyltransferase family. The addition of sialic acid in alpha2,6-linkage to the galactose residue of lactosamine (type 2 chains) is catalyzed by beta-galactoside alpha2,6-sialyltransferase (ST6Gal.I). Although expressed by a single gene, this enzyme shows a complex pattern of regulation which allows its tissue- and stage specific modulation. The cognate oligosaccharide structure, NeuAcalpha2,6Galbeta1,4GlcNAc, is widely distributed among tissues and is involved in biological processes such as the regulation of the immune response and the progression of colon cancer. This review summarizes the current knowledge on the biochemistry of ST6Gal.I and on the functional role of the sialyl-alpha2,6 lactosaminyl structure. PMID- 11425188 TI - A pyrophosphate bridge links the pyruvate-containing secondary cell wall polymer of Paenibacillus alvei CCM 2051 to muramic acid. AB - The peptidoglycan, the secondary cell wall polymer (SCWP), and the surface layer (S-layer) glycoprotein are the major glycosylated cell wall components of Paenibacillus alvei CCM 2051. In this report, the complete structure of the SCWP, its linkage to the peptidoglycan layer, and its physicochemical properties have been investigated. From the combined evidence of chemical and structural analyses together with one- and two-dimensional nuclear magnetic resonance spectroscopy, the following structure of the SCWP-peptidoglycan complex is proposed: [(Pyr4,6) beta-D-ManpNAc-(1-->4)-beta-D-GlcpNAc-(1-->3)]n-11-(Pyr4,6)-beta-D-ManpNAc-(1- >4)-alpha-D-GlcpNAc-(1-->O)-PO2-O-PO2-(O-->6)-MurNAc- Each disaccharide unit is substituted by 4,6-linked pyruvic acid residues. Under mild acidic conditions, up to 50% of them are lost, leaving non-substituted ManNAc residues. The anionic glycan chains constituting the SCWP are randomly linked via pyrophosphate groups to C-6 of muramic acid residues of the peptidoglycan layer. 31P NMR reveals two signals that, as a consequence of micelle formation, experience different line broadening. Therefore, their integral ratio deviates significantly from 1:1. By treatment with ethylenediaminetetraacetic acid, sodium dodecyl sulfate, and sonication immediately prior to NMR measurement, this ratio approaches unity. The reversibility of this behavior corroborates the presence of a pyrophosphate linker in this SCWP-peptidoglycan complex. In addition to the determination of the structure and linkage of the SCWP, a possible scenario for its biological function is discussed. PMID- 11425189 TI - Alkali-catalyzed beta-elimination of periodate-oxidized glycans: a novel method of chemical deglycosylation of mucin gene products in paraffin embedded sections. AB - Altered expression of mucin gene products has been described in many epithelial cancers including colorectal cancer. However, mucins are heavily O-glycosylated making the study of apomucin expression difficult. In this study, we describe a novel method of chemical deglycosylation of mucin gene products on paraffin embedded formalin-fixed tissue sections. In the normal and cancerous colorectum, our results suggest that alkali-catalyzed beta-elimination of periodate oxidized glycan method of chemical deglycosylation modifies the structure of carbohydrates sensitive to mild periodate oxidation resulting in less steric hindrance and selectively removes Tn and sialyl-Tn structures, partially exposing the underlying apomucin epitopes. Using this method, we have demonstrated that the MUC1 tandem repeat epitope recognized by MAb 139H2 is masked predominantly due to steric hindrance by carbohydrate structures whereas the MUC2 tandem repeat epitope recognized by MAb CCP58 and pAb MRP and the MUC3 tandem repeat epitope recognized by pAb M3P are masked by the presence of carbohydrate side chains O linked to Ser/Thr residues within the epitope. Considerable differences in the level and pattern of expression of the epitopes in the tandem repeat region of apomucins of MUC1, MUC2, and MUC3 were observed between normal and cancerous colorectal cancer tissues. We conclude that this novel chemical deglycosylation method that causes selective cleavage of distinct glycans will be useful in unmasking various mucin gene products and glycoproteins containing similar O glycosidic linkages in the tissue sections of formalin-fixed paraffin embedded normal and pathological tissues. PMID- 11425190 TI - Specificity of the binding site of the sialic acid-binding lectin from ovine placenta, deduced from interactions with synthetic analogues. AB - The specificity of the sialic acid-binding lectin from ovine placenta was examined in detail by haemagglutination inhibition assays applying a panel of 32 synthetic sialic acid analogues. The carboxylic acid group is a prerequisite for the interaction with the lectin, the alpha-anomer of the methyl glycoside is only a little more effective as an inhibitor than the beta-anomer and the most potent inhibitor was 9-deoxy-10-carboxylic acid Neu5Ac, followed by 4-oxo-Neu5Ac. In contrast to the majority of known sialic acid-binding lectins, the N-acetyl group of Neu5Ac is not indispensable for binding, neither is the hydroxyl group at C-9 since substitutions at this carbon atom are well tolerated. Furthermore, all sulfur-containing substituents at C-9 enhanced the affinity of the lectin. This is the first sialic acid-binding lectin found to strongly bind thio derivatives. PMID- 11425191 TI - Purification and characterization of a 200 kDa fructosyllysine-specific binding protein from cell membranes of U937 cells. AB - Amadori-modified proteins are bound by macrophages and monocytes via fructosyllysine-specific receptors. Detergent extracts from U937 cell membranes were used to purify the binding proteins by affinity purification on glycated polylysine coated magnetic beads followed by SDS-PAGE. Two proteins of 200 and 100kDa were isolated. MS-analysis of the 200 kDa protein showed high homologies with cellular myosin heavy chain, type A. Both fructosyllysine specific binding proteins, cellular myosin heavy chain and nucleolin, are glycosylated. PMID- 11425192 TI - Monoclonal anti-GD3 antibodies selectively inhibit the proliferation of human malignant glioma cells in vitro. AB - The frequently occurring alteration of ganglioside expression in tumor cells has been implicated to play a role in the uncontrolled growth of these cells; antibodies to such gangliosides might affect tumor cell growth. We have studied the effect of IgM monoclonal antibodies to two glioma-associated gangliosides, GD3 and GM2, on cell proliferation of four human glioma cell lines and one renal tumor cell line. Of the two anti-ganglioside antibodies tested, only the anti-GD3 antibody resulted in a significant (p<0.005) inhibition of cell proliferation as measured by thymidine incorporation and Brd-U labeling, after 24h incubation. The effect was not dependent on any serum factor and no increased cell death was observed. All cell lines contained higher or similar amounts of GM2 than GD3, and both antigens were shown to be expressed on the cell surface and accessible to antibodies. The selective effect of anti-GD3 antibodies as contrasted to the inactivity of anti-GM2 antibodies suggests a possible role for ganglioside GD3 in tumor cell proliferation. PMID- 11425193 TI - Glycoinositolphospholipids from Trypanosoma cruzi induce B cell hyper responsiveness in vivo. AB - The surface of the protozoan Trypanosoma cruzi, the etiologic agent of Chagas' disease, is covered by a dense glycolipid layer, composed mainly by a structurally related family of glycoinositolphospholipids (GIPLs). In the present study we evaluated the in vivo effects of the GIPL on B cell function and immunoglobulin (Ig) secretion. We observed that GIPL injection led to a sustained increase in circulating IgM levels. B cells from GIPL injected mice showed higher response when activated in vitro with either LPS or dextran-conjugated anti-IgD antibodies or purified cytokines. GIPL purified from T. cruzi also showed an adjuvant effect, since this glycophospholipid boosted a polysaccharide-(TNP Ficoll) induced IgG response. Taken together, our data indicate that T. cruzi derived GIPL could be at least partially responsible for the remarkable B cell activation observed during T. cruzi acute infection in vivo. PMID- 11425194 TI - Sialyl-Le(x) and sulfo-sialyl-Le(x) determinants are receptors for P. aeruginosa. AB - Pseudomonas aeruginosa, the main pathogen in the airways of patients suffering from cystic fibrosis (CF), binds to carbohydrate chains of respiratory mucins. Using flow cytometry and polyacrylamide based fluorescent glycoconjugates, it was previously demonstrated that several strains of P. aeruginosa recognize a set of neutral and acidic carbohydrate epitopes found at the periphery of respiratory mucins, especially sialyl-Le(x). This structure, overexpressed in mucins from CF patients, could be responsible in part for the persistence of lung infection in CF patients. The aim of the present work was to determine whether a glycoconjugate bearing the 6-sulfo-sialyl-Le(x) epitope, also found in abundance in CF airway mucins, is also preferentially recognised by different strains of P. aeruginosa. The study was conducted with a nonpiliated strain 1244-NP and four mucoid strains isolated from CF patients. For four strains out of five, the affinity for 6-sulfo-sialyl-Le(x) was as high as for sialyl-Le(x) derivative. These results were confirmed for strain 1244-NP by a microtiter plate assay. PMID- 11425195 TI - The art of diagnosis in head and neck tumors. PMID- 11425196 TI - Selective neck dissection for hypopharyngeal cancer in the clinically negative neck: should it be bilateral? PMID- 11425197 TI - Perilymphatic concentration of gentamicin administered intratympanically in guinea pigs. AB - Intratympanic gentamicin therapy has recently become popular for Meniere's disease, although the administration protocol remains controversial. To date, few studies have been conducted regarding the pharmacokinetics of drugs administered intratympanically. We measured gentamicin concentration in the perilymph close to the round window membrane using a microdialysis technique. A bolus of 0.5 mg gentamicin was put on the round window membrane or a clinically used solution of 5 mg gentamicin was placed in the tympanic bulla of guinea pigs. After 56 +/- 21 min of bolus administration, gentamicin concentration reached a maximum level of 2900 +/- 1200 microg/ml and then decreased, with a half-life of 117 +/- 47 min, characteristic of the kinetics expected with a one-compartment model. With solution administration, the maximum concentration did not differ, but the time to the maximum concentration and the half-life in the perilymph were longer, compared with the results of bolus administration. Our results suggest that even solution administration of gentamicin, which is simple and repeatable, provides a sufficient, stable and continuous supply to the perilymph. PMID- 11425198 TI - Direct evidence of nitric oxide production in the guinea pig organ of Corti. AB - Production of nitric oxide (NO) in the organ of Corti of the guinea pig was investigated using the new fluorescence indicator 4,5-diaminofluorescein diacetate for direct detection of NO. The organ of Corti, lateral wall of the cochlea and isolated outer and inner hair cells were examined to locate NO production sites. The fluorescence intensities were augmented by stimulation with L-arginine or glutamate, and significantly increased after inoculation with lipopolysaccharide. This is the first direct evidence of NO production in the cochlea. NO may play an important role in the physiology of the organ of Corti and may also be involved in hearing disorders. PMID- 11425199 TI - Nitric oxide in guinea pig vestibular sensory cells following gentamicin exposure in vitro. AB - Gentamicin-induced production of nitric oxide (NO) in the vestibular end organs of the guinea pig was investigated using the new fluorescence indicator 4,5 diaminofluorescein diacetate for direct detection of NO. Utricular maculae and isolated vestibular sensory cells were examined to locate NO production sites. The fluorescence intensity of the sensory cells was augmented by stimulation with gentamicin. This increase in fluorescence was inhibited by the presence of the non-specific inhibitor for nitric oxide synthase, L-N(G)-nitroarginine methylester, and by the non-specific N-methyl-D-aspartic acid antagonist (+)-5 methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate. These findings indicate that NO may play an important role in the ototoxicity of aminoglycoside. PMID- 11425200 TI - Methylcobalamin as antagonist to transient ototoxic action of gentamicin. AB - The objective of this study was to determine whether methylcobalamin inhibits the ototoxic action of gentamicin. Fourteen pigmented guinea pigs were randomly divided into experimental and control groups. Each animal in the experimental group was given a large single dose of gentamicin (125 mg/kg i.m.), followed by immediate administration of methylcobalamin (1 mg kg i.m.). In the control group, the animals received gentamicin only, at the same dose as the experimental group. The compound action potentials (CAPs) in the right ears of animals were measured using a signal processor before and 15, 30, 45, 60 and 75 min after injection of the drugs. The sound stimulus was presented by means of clicks and tone pips at 2, 4 and 8 kHz. Following the electro-physiological tests, the animal was sacrificed and prepared for scanning electron microscopic observation. There was prolongation of latency and diminution of amplitude of CAPs N1 and N2 15 min after injection of gentamicin. Although the N1-N2 intervals increased as the time after injection of gentamicin increased, the variations were still within normal ranges. In contrast, there was no prolongation of N1 latencies or N1-N2 intervals 30 min after injection in the experimental group. There were no marked changes in N1 or N2 amplitudes, compared with the results before injection, in the experimental group. The firing mechanism of the inner ear may be affected by a single dose of gentamicin. This suggests that the transient ototoxic action of gentamicin hardly affected the conductive function of the nerve fibres, even though the firing function of the inner ear was influenced. Methylcobalamin may inhibit the ototoxic side effects of gentamicin. PMID- 11425201 TI - Comparison and evolution of the lagena in various animal species. AB - The structure of the vestibular organs of the teleost fish (bluegill), newts (Japanese fire-belly newt), frogs (black-spotted pond frog), snapping turtles and birds (chicks) was morphologically compared, with particular attention to the lagena macula, and the differences between animal species with relation to evolution were considered. Teleost fish had no striola on the lagena macula. The striola of newts were short and restricted to the central area of the macula, but those of frogs, snapping turtles and chicks extended from the anterior to posterior edges of the macula. This indicates that the frog is more highly evolved than the newt. The length of the kinocilium of sensory hairs was equal to that of the longest stereocilium in teleost fish and newts, but the kinocilia of frogs, snapping turtles and chicks were longer than the longest stereocilium. This indicates that the function of the lagena of teleost fish and newts is for hearing whilst in the other animals they are for posture. The diameter of the sensory hair bundles is small in teleost fish and frogs, but large in newts and snapping turtles. This indicates that the sensitivity of the sensory cells of the lagena towards outer force is low in teleost fish and frogs, high in newts and snapping turtles and intermediate in birds. The lagena of snapping turtles protrudes from the basilar papilla into the vestibule but the lagena of chicks lies on the tip of the long projecting basilar papilla. From observation of the locations of lagenae it is natural to speculate that there must have been some species of animal now extinct that had the evolving location of the lagena prior to that of chicks. In future it will be very interesting and useful to identify this extinct animal using DNA techniques. PMID- 11425202 TI - Up-regulation of MUC5AC mRNA expression in endotoxin-induced otitis media. AB - We investigated the expression levels of MUC5AC in endotoxin-induced otitis media with effusion (OME) in the rat using competitive polymerase chain reaction (PCR) and the morphology of middle ear mucosa using transmission electron microscopy (TEM). Experimental OME in the rat was induced after middle ear instillation of Escherichia coli lipopolysaccharides (LPS). Middle ear mucosa were obtained at 0 h, 12 h, Day 1, Day 3, Day 7 and Day 14 and reverse transcriptase (RT)-PCRs were then performed for the identification of MUC1, MUC2, MUC5AC and submandibular mucin 1 expression, followed by competitive PCRs for MUC5AC and beta2 microglobulin expression. Normal middle ear mucosa revealed no expression of mucin genes, whereas endotoxin upregulated the expression of MUC5AC mRNA between 12 h and Day 7, with maximal expression at Days 1 and 3. Middle ears treated three times with LPS upregulated more MUC5AC mRNA expression, by a factor of approximately 3.5, than those 1 day after one instillation. On TEM, dark granulated cells were observed at Day 3 after endotoxin instillation, but mixed granulated cells were seen on the ears treated three times with LPS. These results suggest that MUC5AC could be one of the major mucin genes in the middle ear mucosa related to otitis media. PMID- 11425203 TI - Production of nitric oxide by the middle ear epithelium and subsequent inhibition of sodium transport. AB - The production of nitric oxide (NO) within the middle ear has not previously been characterized. The presence of NO synthase (NOS) transcripts was demonstrated using RNA amplification by reverse transcriptase-polymerase chain reaction in rat middle ear mucosa and in rat primary cultured middle ear epithelial cells. The expression of NOS was indirectly assessed by nitrite measurement in the supernatant of primary cultured cells. The effect of NO on ion transport was investigated in a previously described middle ear epithelial cell line using the short-circuit current (Isc) technique. NO per se had no effect on Isc. However, previous work has shown that sodium transport is stimulated by reactive oxygen species (ROS). NO blunted this stimulation, an inhibition probably related to the toxicity of peroxynitrite, ONOO-, a highly reactive compound. These results suggest that NO is produced by middle ear epithelial cells and that, in the presence of ROS, NO may be responsible for an inhibition of ion transport viaperoxynitrite formation. PMID- 11425204 TI - Auditory pathway function after vestibular schwannoma surgery. AB - We studied seven patients before and after vestibular schwannoma surgery. Four patients became unilaterally profoundly deaf and three patients preserved their hearing. Cortical responses were recorded with a 122-channel whole-scalp SQUID neuromagnetometer using tone-burst stimuli to the healthy ear. Brainstem auditory evoked potentials (BAEPs) were measured using alternating clicks. Ten healthy volunteers served as a control group. In patients, preoperative cortical response latencies and strengths did not differ significantly from those of controls. However, 6 months after the operation the latency was, on average, 7 ms longer than preoperatively over both hemispheres. BAEPs were in the normal range both before and after the operation. These results suggest that unilateral lesion in peripheral auditory pathways also affects cortical reactivity to stimuli presented to the non-affected ear, possibly reflecting altered binaural interaction in the auditory pathways. PMID- 11425205 TI - A post-processing protocol for three-dimensional visualization of the inner ear using the volume-rendering technique based on a standard magnetic resonance imaging protocol. AB - A superior diagnostic quality compared to other post-processing (PP) techniques for three-dimensional (3D) inner ear imaging has been attributed to volume rendering (VR). We defined and assessed a VR protocol for 3D visualization of the inner ear in a routine imaging setting. Following definition of a VR protocol by using lower threshold values, surface shading, perspective views and related parameters, standardized 3D views of the inner ear were generated and evaluated in 32 patients suffering from sensorineural or combined hearing loss. Comprehensive inner ear visualization was achieved in 28 patients by means of two 3D shaded-surface views. Incomplete data acquisition (1/32), motion and pulsation artefacts (2/32) and interposed fluid-retaining mastoid cells (1/32) were responsible for non-diagnostic image quality in 4/32 patients. In 5/32 patients modifications of the PP protocol involving the threshold value and depth-cueing parameters helped to establish diagnostic image quality. Mean post-processing time amounted to 5.8 min per site. 3D imaging with the VR technique is suitable for routine inner ear assessment if direct VR, predefined PP protocols and standardized labyrinthine views are used. PMID- 11425206 TI - Intratympanic therapy for Meniere's disease: effect of administration of low concentration of gentamicin. AB - We present results at 2 years follow-up of a group of 15 patients with Meniere's disease treated with a low concentration of intratympanic gentamicin (IG group), compared with a group of 15 patients who refused any surgical treatment (NH group). IG was administered according to a predetermined and fixed schedule consisting of 2 doses of 0.5 ml gentamicin solution, pH 7.8, injected once a week, with a drug concentration of 20 mg/ml; the total dose of gentamicin was < or = 20 mg. Additional doses of IG were administered only in patients who had recurrence of vertigo. The results were evaluated following the American Academy of Otolaryngology Head and Neck Surgery 1995 criteria for reporting Meniere's disease treatment results by means of interviews, audiologic and vestibular evaluations, and a questionnaire based on a six-point functional level scale. Seven of the 15 IG patients had recurrence of vertigo after the second injection and received a third dose of IG. Four patients had recurrence of vertigo after the third infiltration; three subjects received a fourth dose and one refused additional injection. At 2 years follow-up, 93% of the IG patients had complete (class A) or substantial (class B) control of vertigo. Only 47% of the NH patients had no vertigo or were substantially improved. Hearing deteriorated in 7% of the IG group and in 40% of the NH group. Tinnitus disappeared or improved in 20% of the IG patients and in 27% of the NH patients; 40% of the IG patients and 27% of the NH patients reported that their aural pressure was abolished. The present study demonstrates that, in patients with Meniere's disease, 0.5 ml doses of gentamicin solution, with a concentration of 20 mg/ml, injected intratympanically once a week minimize the risk of hearing loss in the treated ear, permitting complete control of vertigo in more than half of cases after 2 doses and in almost all subjects (93%) after 4 doses. PMID- 11425207 TI - Effects of cadmium on the hearing system. AB - The functional resemblance between kidney proximal tubular and inner ear epithelial cells which has often been pointed out in the literature led us to hypothesize that nephrotoxic agents that cause renal tubular injury might also impair the function of inner ear cells. As one of the most toxic environmental nephrotoxic agents is cadmium, we aimed to study its effects on hearing experimentally in rats. In this study, increased blood and renal cortical cadmium levels were associated with high cadmium accumulation in ear ossicles and labyrinth in rats exposed to cadmium. The changes in auditory brainstem response (ABR) and otoacoustic emission in 2-month-old male rats exposed to drinking water containing 5 and 15 ppm CdCl2 for 30 days showed that cadmium-induced nephrotoxicity was associated with signs of defective hearing at a concentration of 15 ppm CdCl2 but that 5 ppm CdCl2 caused hearing loss without affecting kidney function. The mean latency of ABR wave 1, which indicates the function of the cochlea, was 1.335 +/- 0.31 ms in the control group and 1.641 +/- 0.052 and 1.74 +/- 0.88 ms in the rats subjected to 5 and 15 ppm CdCl2, respectively (p < 0.001). In the cadmium-treated groups short interpeak wave I-III latencies (p < 0.01) indicated cochlear dysfunction and this was also supported by the distortion product otoacoustic emission results (p < 0.001). Non-significant changes in wave III and V latencies were accepted as evidence of unaltered function of the other parts of the auditory system. These results suggest that hair cells are more sensitive to cadmium than kidney tubule cells and that the cochlear component of hearing is more vulnerable to cadmium toxicity than other parts of the auditory system. PMID- 11425208 TI - Analysis of proteoglycan gene messages in human nasal mucosa and nasal polyp using dot blot hybridization. AB - It has been suggested that the formation and growth of nasal polyp require the remodeling of extracellular matrix. Proteoglycans (PGs) are major components of the extracellular matrix that maintain the integrity of structural tissue. The leucine-rich repeat PGs include lumican, decorin and biglycan and have many important biologic activities in various pathologic conditions, including the remodeling of the extracellular matrix. Therefore, these small-PG families may be involved in the formation and growth of nasal polyp. In the present study, surgical specimens of nasal polyps and nasal mucosa were assessed for expression of mRNA coding for lumican, decorin and biglycan using reverse transcriptase polymerase chain reaction followed by dot blot hybridization. Lumican, decorin and biglycan mRNA were expressed in all tissue samples examined. Semiquantitative dot blot hybridization revealed that the levels of the lumican and biglycan messages are lower in nasal polyp tissues than in nasal mucosa. The decorin messages in nasal polyp were expressed at levels similar to those in nasal mucosa. These results suggest that lumican, decorin and biglycan may be important components of the extracellular matrix in nasal mucosa. Considering the function of these PGs, normal levels of decorin associated with low levels of biglycan and lumican may play a role in the pathogenesis of nasal polyposis. PMID- 11425209 TI - Expression of 5-HT1B receptors in human nasal mucosa. AB - 5-HT1B receptors were discovered in human nasal mucosa using immunocytochemistry. Strong immunoreaction was seen around small blood vessels mainly confined to the smooth muscle cell layer. In contrast, no immunoreaction for 5-HT1D was seen. The possibility of local release in connection with specific target receptors suggests a role for 5-HT in the regulation of vascular tone, glandular secretion and epithelial functions and that 5-HT1B receptor agonists may be of clinical importance. PMID- 11425210 TI - Dual symptomatic and exudative nasal responses are not characteristics of perennial allergic rhinitis. AB - Acute and late phase (dual) symptomatic responses after allergen challenge commonly occur in allergic asthma. The aim of the present study was to explore the occurrence of allergen challenge-induced biphasic responses in patients with chronic perennial allergic rhinitis living in an area with high house dust mite (HDM) exposure. Fifteen patients with perennial rhinitis and evident allergy to HDM participated. Nasal challenges with HDM and sham were performed on separate days in a crossover design. Nasal symptoms, forced expiratory volume in 1 s (FEV1) and orally exhaled nitric oxide were recorded every hour for up to 8 h after each challenge. Alpha2-macroglobulin and eosinophil cationic protein (ECP) were analysed in hourly nasal lavages, and nasal histamine provocations were carried out after 8 h. HDM, but not sham, caused an immediate increase in nasal symptoms which gradually abated over the 8-h period. No reoccurrence of nasal late phase symptoms was seen. HDM, but not sham, induced an early increase in alpha2-macroglobulin and ECP levels: both indices remained elevated for up to 3 h after challenge. HDM challenge evoked hyper-responsiveness to histamine expressed as increased nasal symptoms (p < 0.05; HDM vs sham). No differences in exhaled nitric oxide or FEV1 were demonstrated at any one time point between the HDM- and sham-challenged days. It is concluded that nasal symptomatic and exudative late phase responses may not be a general feature, even in subjects with perennial rhinitis challenged with high, symptom-provoking, doses of HDM allergen. PMID- 11425211 TI - Expression of human beta-defensin 1 mRNA in human palatine tonsil. AB - Defensins are a newly delineated family of effector molecules whose contribution to host defense, inflammation and cytotoxicity may be considerable for humans. Beta-defensins are cationic peptides with broad-spectrum antimicrobial activity that are produced by epithelia at the mucosal surface. Tonsillar epithelium, which is constantly exposed to microorganisms, may express these natural antibiotic peptides as part of its protective function. In this study we searched for the expression of hBD-1 mRNA in palatine tonsillar epithelium without signs of infection, using reverse transcriptase-polymerase chain reaction and in situ hybridization. Total RNA was isolated from non-infected tonsil and hBD-1 mRNA was identified in these tissues. From in situ hybridization, the expression of hBD-1 mRNA was seen to be localized in the surface epithelia of palatine tonsil. These data suggest that hBD-1 of the palatine tonsil may also play an important role in innate defense against microorganisms. PMID- 11425212 TI - Lateral medullary infarct presenting as acute dysphagia. AB - Lateral medullary infarct (LMI) usually presents with a variety of neurological features. We describe a case of LMI in which acute dysphagia was the only initial symptom. This typical neurological syndrome is very unlikely to be found during ENT practice; however, it needs to be considered among the differential diagnoses when encountering dysphagia of uncertain origin. PMID- 11425213 TI - The sentinel node procedure with Patent Blue V dye in the surgical treatment of papillary thyroid carcinoma. AB - How far to extend the surgical treatment of papillary thyroid carcinoma (PTC) is still an open question. A contribution may come from intra-operative lymphatic mapping because, in other malignancies, the procedure has become an important aid in defining lymph node status. To assess the feasibility of using the sentinel lymph node (SLN) technique with the intratumoral injection of Patent Blue V dye to guide nodal dissection in PTC, 29 patients with a preoperative diagnosis of PTC and no clinical or ultrasonographic evidence of nodal involvement underwent cervicotomy and exposure of the thyroid gland, followed by Patent Blue V dye injection into the thyroid nodule. Total thyroidectomy was subsequently performed, resecting the lymph nodes at levels III, IV, VI and VII. The thyroid, SLN and the other lymph nodes were snap-frozen and submitted for both intra operative and subsequent definitive pathological evaluation. Intra-operative lymphatic mapping located the SLN in 22/29 patients (75.9%) and the SLN revealed neoplastic involvement in 4/22 (18.2%); other lymph nodes were also positive in 2 cases. In the 18 patients whose SLNs were not metastatic, the other nodes were also disease-free. The SLN technique thus seems helpful in avoiding unnecessary lymph node dissection in PTC without spread to the SLN. PMID- 11425214 TI - Free flap reconstructions in the management of oral and pharyngeal cancer. AB - Fifty patients who had undergone microvascular free flap reconstruction of the oral cavity or pharynx for malignancy between 1989 and 1995 were retrospectively analysed to find factors predicting postoperative complications and outcomes. The mean follow-up time was 2.6 years. More than half (26/50) of the patients had a stage IV malignancy and 10 patients had a recurrent tumour. The overall flap success rate, with 2 flap losses, was 96%. Postoperative medical complications occurred in 29/50 (58%) cases. The recipient site of 10/50 (20%) flaps required re-exploration. Mortality was 2%, with 1 death occurring within 30 days. The mean survival rate after the microvascular operation was 1.6 years, and the 3- and 5 year survival rates were 42% and 23%, respectively. Patients with an oropharyngeal tumour seemed to have the best prognosis and those with a hypopharyngeal tumour the poorest. Men had a threefold greater risk of dying within < 1 year postoperatively compared with women. Tumour stage was the strongest attribute influencing survival. The risk of death after the microvascular procedure increased 2.8-fold when the stage advanced from II to III or from II to IV. None of the tested variables alone could predict complications. PMID- 11425215 TI - Effect of strong detergents and chaotropes on the detection of proteins in two dimensional gels. AB - The solubilization of a particular protein is mandatory for its subsequent resolution and detection in two-dimensional gels. However, the extraction solutions, that are compatible with the first-dimensional separation step, such as urea and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), do not solubilize all proteins in a sample. We studied the effect of various common, strong detergents and chaotropes, widely used as solubilizing agents, such as sodium dodecyl sulfate, lithium dodecyl sulfate and guanidine hydrochloride, on the solubilization of the total and membrane proteins of the bacterium Haemophilus influenzae. The proteins solubilized with each system were analyzed by two-dimensional electrophoresis and these of interest were identified by matrix assisted laser desorption/ionization-mass spectrometry (MALDI-MS). Use of sodium dodecyl sulfate, lithium dodecyl sulfate or guanidine hydrochloride for the solubilization of total proteins of the microorganism resulted in the detection of several additional spots, representing mainly outer membrane proteins, in comparison with those detected in the soluble protein fraction. Solubilization of the proteins of the cell envelope fraction with sodium dodecyl sulfate did not result in a more efficient protein detection when compared to the extraction with the urea/CHAPS system. When the dry immobilized pH gradient strips were rehydrated in a solution containing the proteins of the membrane fraction solubilized with sodium dodecyl sulfate or lithium dodecyl sulfate, a larger number of protein spots were detected in comparison with strips that were rehydrated in the urea/CHAPS solution. However, no improvement was observed in comparison with protein application in sample cups. The additional proteins detected with the use of strong detergents and chaotropes are in the majority difficult to solubilize and less hydrophobic proteins. PMID- 11425216 TI - Towards global analysis of mammalian proteomes using sample prefractionation prior to narrow pH range two-dimensional gels and using one-dimensional gels for insoluble and large proteins. AB - The number of unique protein species in proteomes from a single mammalian cell type is not well defined but is likely to be at least 10000-20000. Since standard size two-dimensional gels typically resolve only about 1500 to 3000 spots, they merely analyze a small portion of these proteomes. In addition, all insoluble proteins and typically proteins > 100 kDa are seldom resolved on two-dimensional (2-D) gels. The current study demonstrates the feasibility of an overall strategy for more comprehensive quantitative comparisons of complex proteomes derived from physiological fluids or mammalian cell extracts. A key feature of this approach is to prefractionate samples into a few well-resolved fractions based on the proteins' isoelectric points (pIs) using microscale solution isoelectric focusing. These fractions are then separated on narrow pH range two-dimensional gels approximately +/- 0.1 pH unit wider than the prefractionated pool. When this prefractionation approach is applied to complex mammalian proteomes, it improves resolution and spot recovery at high protein loads compared with use of parallel narrow pH range gels without prefractionation. The minimal cross-contamination between fractions allows quantitative comparisons in contrast to most alternative prefractionation methods. In addition, complementary data can be obtained by parallel analysis of the solubilized fraction on high-resolution large-pore gradient one-dimensional gels followed by mass spectrometric identification to analyze proteins between 100 and approximately 500 kDa. Similarly, insoluble proteins can be analyzed using large-pore gels for large proteins and 10-12% one dimensional sodium dodecyl sulfate (SDS) gels for smaller proteins. Together, these strategies should permit more reliable quantitative comparisons of complex mammalian proteomes where detection of at least 10000 protein spots is needed in order to analyze the majority of the unique protein species. PMID- 11425217 TI - Automatic registration for images of two-dimensional protein gels. AB - Two-dimensional polyacrylamide gel electrophoresis 1 (2-D PAGE 1) is currently the method of choice for separating complex mixtures of cellular proteins. Despite its usefulness, 2-D PAGE is not being applied to its full potential because of difficulties with both the method and analysis of the results. One of the key problems is the difficulty and slowness of image analysis, especially registration (image alignment), which is laborious and the results unsatisfactory. We have developed a novel system for analysis of 2-D PAGE images, called Z3, that performs the analysis faster and more precisely. The Z3 system employs novel approaches to image registration, image display, computation of differential expression, and the design and analysis of 2-D gel experiments. This paper describes in detail the registration algorithm, and briefly discusses the merits of complementary pseudocolor display. The registration algorithm is novel in that for the first time raw-image-based registration technology is applied to 2-D gel analysis. PMID- 11425218 TI - The scaled volume as an image analysis variable for detecting changes in protein expression levels by silver stain. AB - A new variable for measuring the relative intensities of silver stained protein spots on two-dimensional gels is described. The scaled volume (SV) more accurately measures the intensity of protein spots and accounts for differences frequently encountered when trying to compare two gels than other variables such as relative volume ratio, optical density, or relative optical density. The SV scales the signal of interest by the noise (gel background) with secondary signals removed (spots not of interest, e.g., technical artifacts). The SV of spot intensities offers a better dynamic response to protein amount for the model proteins studied here. Depending on the quantity of protein loaded onto gels, we have observed a coefficient of variation range of 0.2 to 1.3. We also observe that the SV silver stain response follows a characteristic exponential profile for different proteins. PMID- 11425219 TI - Protein alkylation by acrylamide, its N-substituted derivatives and cross-linkers and its relevance to proteomics: a matrix assisted laser desorption/ionization time of flight-mass spectrometry study. AB - The present review highlights some important alkylation pathways of proteins, as measured by matrix assisted laser desorption/ionization-time of flight (MALDI TOF)-mass spectrometric analysis, engendered by acrylamide and a number of its derivatives, including N-substituted acrylamides, cross-linkers and Immobilines (the acrylamido weak acids and bases used to create immobilized pH gradients). The present data are of relevance in two-dimensional maps and proteome analysis. It is shown that acrylamide can alkylate the -SH group of proteins even when engaged in disulfide bridges. An order of reactivity is obtained for a series of cross-linkers, which are shown to have an extremely reacting double bond, with the second one almost unreactive, originating "pendant, unreacted ends", which can subtract proteins migrating in a gel by covalently affixing them to it. An analogous reactivity scale is constructed also for the Immobiline chemicals, whose reactivity is shown to be linearly dependent on the pK values, the least reacting species being the acidic compounds. When analyzing real-life samples by two-dimensional (2-D) maps, like milk powders, a number of modifications can be detected by MALDI-TOF mass spectra of eluted spots, including variable phosphorylation sites (up to nine) and lactosyl moieties. If, for eluting such spots, formic acid is used, MALDI-TOF mass spectrometry (MS) reveals an incredible number of formylation sites, on Ser and Thr residues. PMID- 11425220 TI - Protein identification based on matrix assisted laser desorption/ionization-post source decay-mass spectrometry. AB - Due to its very short analysis time, its high sensitivity and ease of automation, matrix-assisted laser desorption/ionization (MALDI)-peptide mass fingerprinting has become the preferred method for identifying proteins of which the sequences are available in databases. However, many protein samples cannot be unambiguously identified by exclusively using their peptide mass fingerprints (e.g., protein mixtures, heavily posttranslationally modified proteins and small proteins). In these cases, additional sequence information is needed and one of the obvious choices when working with MALDI-mass spectrometry (MS) is to choose for post source decay (PSD) analysis on selected peptides. This can be performed on the same sample which is used for peptide mass fingerprinting. Although in this type of peptide analysis, fragmentation yields are very low and PSD spectra are often very difficult to interpret manually, we here report upon our five years of experience with the use of PSD spectra for protein identification in sequence (protein or expressed sequence tag (EST)) databases. The combination of peptide mass fingerprinting and PSD and analysis described here generally leads to unambiguous protein identification in the amount of material range generally encountered in most proteome studies. PMID- 11425221 TI - Rapid quantitative measurements of proteomes by Fourier transform ion cyclotron resonance mass spectrometry. AB - The patterns of gene expression, post-translational modifications, protein/biomolecular interactions, and how these may be affected by changes in the environment, cannot be accurately predicted from DNA sequences. Approaches for proteome characterization are generally based upon mass spectrometric analysis of in-gel digested two dimensional polyacrylamide gel electrophoresis (2 D PAGE) separated proteins, allowing relatively rapid protein identification compared to conventional approaches. This technique, however, is constrained by the speed of the 2-D PAGE separations, the sensitivity limits intrinsic to staining necessary for protein visualization, the speed and sensitivity of subsequent mass spectrometric analyses for identification, and the limited ability for accurate quantitative measurements based on differences in spot intensity. We are presently developing alternative approaches for proteomics based upon the combination of fast capillary electrophoresis, or other suitable chromatographic separations, and the high mass accuracy and sensitivity obtainable with unique Fourier transform ion cyclotron resonance (FTICR) mass spectrometers available at our laboratory. Several approaches are presently being pursued; one based upon the analysis of intact proteins and the second upon approaches for global protein digestion and accurate peptide mass analysis. Quantitation of protein/peptide levels are based on using two or more stable isotope labeled versions of proteomes which are combined to obtain precise quantitation of relative protein abundances. We describe the status of our efforts towards the development of a high-throughput proteomics capability and present initial results for application to several microorganisms and discuss our efforts for extending the developed capability to mammalian proteomes. PMID- 11425222 TI - Identification of post-translationally modified proteins in proteome studies. AB - Proteome studies are powerful tools to solve many different problems in metabolism, signal transduction, drug discovery, and other areas of interest in life sciences. Up to now, high-sensitive methods for protein identification after two-dimensional gel electrophoresis using mass spectrometry are available. However, the identification of post-translational modifications after two dimensional gel electrophoresis is still an unsolved problem. In this paper, we want to give several examples for the successful identification of post translational modifications and point mutations. PMID- 11425223 TI - An optimized procedure for detection of proteins on carrier ampholyte isoelectric focusing and immobilized pH gradient gels with imidazole and zinc salts: its application to the identification of isoelectric focusing separated isoforms by in-gel proteolysis and mass spectrometry analysis. AB - A method for the characterization of proteins separated by isoelectric focusing in carrier ampholytes (CA-IEF) or immobilized pH gradient (IPG) gels by in-gel digestion and mass spectrometry is described. Proteins are detected by an improved imidazole-Sodium dodecyl sulfate (SDS)-zinc staining adapted for IEF and IPG gels. Sensitivity is close to that of mass spectrometry-compatible silver staining, but simpler and faster. Proteins were digested in imidazole-SDS-zinc stained CA-IEF and IPG gels in the presence of a zinc-chelating agent. Mass spectra were clearly interpretable as carrier ampholytes which were efficiently removed before digestion; high-sequence coverage that allowed isoform characterization was obtained by analyzing both the aqueous and the organic phase extracts. PMID- 11425224 TI - Two-dimensional electrophoresis and peptide mass fingerprinting of bacterial outer membrane proteins. AB - Many bacterial outer membrane proteins (OMPs) are missing from two-dimensional (2 D) gel proteome maps. Recently, we developed a technique for 2-D electrophoresis (2-DE) of Escherichia coli OMPs using alkaline pH incubation for isolation of OMPs, followed by improved solubilization conditions for array by 2-DE using immobilized pH gradients. In this report, we expanded our study, examining protein components from the outer membranes of two enteric bacteria, Salmonella typhimurium and Klebsiella pneumoniae (also known as Klebsiella aerogenes), as well as the unrelated, free-living alpha-proteobacteria Caulobacter crescentus. Patterns of OMPs expression appeared remarkably conserved between members of the Enterobacteriaceae, while C. crescentus was unique, displaying a greater number of clusters of higher-molecular-weight proteins (>80 kDa). Peptide mass fingerprinting (PMF) was used for protein identification, and despite matching across-species boundaries, proved useful for first-pass protein assignment of enteric OMPs. In contrast, identification of C. crescentus OMPs was successful only when searching against its recently completed genome. For all three microorganisms examined, the majority of proteins identified on the 2-D gel appear localized to the outer membrane, a result consistent with our previous finding in Escherichia coli. In addition, we discuss some of the benefits and limitations of PMF in cross-species searching. PMID- 11425225 TI - Time-dependent expression and processing of a hypothetical protein of possible importance for regulation of the Chlamydia pneumoniae developmental cycle. AB - Chlamydia pneumoniae is an obligate intracellular human pathogen infecting epithelial cells of the upper respiratory tract. It is a Gram-negative bacteria and has a unique biphasic developmental cycle. In this study, we use two dimensional gel electrophoresis in combination with radioactive labeling to investigate time-dependent expression and processing of C. pneumoniae proteins. We report on (i) the identification of a hypothetical protein which is expressed late in the developmental cycle and subsequently processed; we speculate that this protein may be of importance for the developmental cycle of Chlamydia; (ii) the identification of the major outer membrane protein in three different variants, which may all be present in vivo. PMID- 11425226 TI - Media containing aromatic compounds induce peculiar proteins in Acinetobacter radioresistens, as revealed by proteome analysis. AB - An Acinetobacter radioresistens strain able to grow on phenol or benzoate as sole carbon and energy source through the beta-ketoadipate pathway was isolated in our laboratories. In previous research, we found a different expression of catechol 1,2-dioxygenase isoenzymes (C-1,2-O) depending on the growth substrate (phenol or benzoate). In the present study, we used proteome techniques to extend our investigation to other enzymes involved in the aromatic degradation pathway. Since the first nontoxic metabolite in this route is cis,cis-muconic acid, we focused our attention on the enzymes leading to this compound, chiefly phenol hydroxylase (PH), benzoate dioxygenase (BD), cis-1,2-dihydroxycyclohexa-3,5-diene 1-carboxylate dehydrogenase (D) and C-1,2-O. In particular, the A. radioresistens proteome was monitored under different growth substrate conditions, using acetate, benzoate, or phenol as sole carbon source. We compared the protein maps by software image analysis and detected marked differences, suggesting the inducibility of most enzymes. This research also sought to evaluate the conditions allowing the best expression of enzymes to be used in immobilized systems suitable for bioremediation. The experimental data indicate that benzoate is the best carbon source to gain the highest amount of C-1,2-O and D, while phenol is the best growth substrate to obtain PH. PMID- 11425227 TI - Proteome analysis of Corynebacterium glutamicum. AB - By the use of different Corynebacterium glutamicum strains more than 1.4 million tons of amino acids, mainly L-glutamate and L-lysine, are produced per year. A project was started recently to elucidate the complete DNA sequence of this bacterium. In this communication we describe an approach to analyze the C. glutamicum proteome, based on this genetic information, by a combination of two dimensional (2-D) gel electrophoresis and protein identification via microsequencing or mass spectrometry. We used these techniques to resolve proteins of C. glutamicum with the aim to establish 2-D protein maps as a tool for basic microbiology and for strain improvement. In order to analyze the C. glutamicum proteome, methods were established to fractionate the C. glutamicum proteins according to functional entities, i.e., cytoplasm, membranes, and cell wall. Protein spots of the cytoplasmic and membrane fraction were identified by N terminal sequencing, immunodetection, matrix assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) and electrospray ionization-mass spectrometry (ESI-MS). Additionally, a protocol to analyze proteins secreted by C. glutamicum was established. Approximately 40 protein spots were observed on silver-stained 2-D gels, 12 of which were identified. PMID- 11425228 TI - The proteome of maize leaves: use of gene sequences and expressed sequence tag data for identification of proteins with peptide mass fingerprints. AB - As a first step in establishing a proteome database for maize, we have embarked on the identification of the leaf proteins resolved on two-dimensional (2-D) gels. We detected nearly 900 spots on the gels with a pH 4-7 gradient and over 200 spots on the gels with a pH 6-11 gradient when the proteins were visualized with colloidal Coomassie blue. Peptide mass fingerprints for 300 protein spots were obtained with matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometer and 149 protein spots were identified using the protein databases. We also searched the pdbEST databases to identify the leaf proteins and verified 66% of the protein spots that had been identified using the protein databases. Sixty-seven additional protein spots were identified from expressed sequence tags (ESTs). Many abundant leaf proteins are present in multiple spots. Functions of over 50% of the abundant leaf proteins are either unknown or hypothetical. Our results show that EST databases in conjunction with peptide mass fingerprints can be used for identifying proteins from organisms with incomplete genome sequence information. PMID- 11425229 TI - Identification of putative proteins involved in granule biogenesis of tick salivary glands. AB - Ticks secrete bioactive components during feeding that assist them in gaining a blood meal. Compounds secreted are stored in granules until a stimulus induces secretion during feeding. Biogenesis of tick secretory granules has not been investigated before. An adequate understanding of granule biogenesis could advance our understanding of tick salivary gland biology and could aid in the rational design of tick control methods. Putative tick salivary gland proteins 1 4 (TSGP1-4) involved in granule biogenesis were identified in this study based on their abundance in salivary gland extracts and granule preparations and their ability to aggregate under conditions of slight acidity and high calcium concentration. TSGP2 and TSGP3 have been identified as previously described toxic and nontoxic homologues, respectively, while toxicity was also associated with TSGP4. PMID- 11425230 TI - Two-dimensional database of mouse liver proteins. An update. AB - We updated the two-dimensional protein database for mouse liver. Microsomal and cytosolic fractions of the liver proteins from male mice were separated by two dimensional electrophoresis. The proteins were identified by Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) on the basis of peptide mass fingerprinting, following in-gel digestion with trypsin and matching with the theoretical peptide masses of all known proteins from all species. Approximately 5800 spots, excised from 14 two-dimensional gels, were analyzed which resulted in the identification of about 2500 proteins that were the products of 328 different genes. The database includes 112 newly identified gene products. The fractionation prior to two-dimensional electrophoresis was essential for the detection of the new proteins, 55% of which were found in the microsomal and 35% in the cytosolic fraction. The more frequently identified proteins in the various gels were heat shock proteins, house-keeping enzymes, such as ATP synthase chains, disulfide isomerase, and structural proteins, such as tropomyosin. About 45% of the identified proteins were detected 1-3 times, 45% 4-9 times, and the rest 10 or more times. Most proteins were represented by many spots. In average, about 18-20 spots were detected per gene product. PMID- 11425231 TI - Protein profiles of the Chinese hamster ovary cells in the resting and proliferating stages. AB - Identification and characterization of the proteins that regulate the transition from the resting stage (G0) through G1 to S phase of the cell cycle are of central importance to understand the control of cell proliferation and chromosome replication. Unlike in lower organisms, where relatively small numbers of key factors are involved in this process, the factors involved in the same control mechanisms in mammalian systems are much more complex. Furthermore, accumulating lines of evidence now suggest that the nuclear matrix and chromatin organization also play an essential role for the cell cycle control in mammalian cells. To gain a better understanding of the overall dynamics and changes of the protein factors in the context of matrix/chromatin organization, we examined the protein profiles of the Chinese hamster ovary (CHO) cells in different cell cycle compartments. The methods used in this study included subcellular fractionations (cytosol, nuclear extraction, chromatin, and nuclear matrix), two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), silver staining, and immunoblotting. As expected, significant changes of protein profiles were observed when cells entered into proliferating stages from G0. Among approximately 1200 protein spots analyzed by 2-D PAGE, at least 12 showed marked increase or decrease at this transitional period. Further cell-cycle progression from G1 to S phase showed less dramatic changes of overall protein protile. However, the profile of certain proteins showed rather dramatic changes of their subcellular localization during this transitional period. In particular, the levels of proliferating cell nuclear antigen (PCNA) in the nuclear matrix and chromatin dramatically increased in mid-G1 and in the beginning of S phase, respectively, while the overall PCNA level was relatively constant throughout the cell cycle. PMID- 11425232 TI - A proteomic approach to mimic fibrosis disease evolvement by an in vitro cell line. AB - Subepithelial fibrosis in asthma involves an increase in the thickening of the lamina reticularis and is due to increased deposition of collagen I, III and V, and fibronectin. The cause of the thickening of the reticular layer is not known in detail, however, it is proposed to be caused by bronchial myofibroblasts. The transformation of fibroblasts to myofibroblasts may be contributed by inflammatory cytokines. In this paper we have studied and compared in vivo tissue material with a human fibroblast target cell. A normal primary fetal fibroblast cell line and HFL-1 (human fibroblast lurg cells) were used as a comparison between fibroblasts from human central biopsies regarding morphology and cell proliferation. Both cell morphology and cell proliferation rate was similar between the different set of cell cultures. Furthermore, it could be concluded that fibroblasts cultures from patients with asthma were surrounded by more extracellular matrix molecules compared to the primary cell line HFL-1, which may mimic the in vivo situation during formation of fibrosis. We wanted to investigate if differential protein display by two-dimensional (2-D) gel electrophoresis and subsequent protein identification by matrix assisted laser desorption/ionization-time of flight (MALDI-TOF)-mass spectrometry could reveal proteins induced by cytokine stimulation that can be correlated to the transformation of normal human fetal lungs cells into a more myofibroblast like phenotype. After stimulation with transforming growth factor-beta (TGF-beta) several myofibroblast markers were found to be regulated. Especially cytoskeletal and cytoskeletal-associated proteins like actin isoforms and tropomyosin, proteins that are responsible for contraction as well as transportation of extra cellular matrix proteins, which are overproduced in the formation of fibrosis. These results indicate that TGF-beta, which is increased in a fibrotic process, participates in the transformation of fibroblasts to myofibroblasts. PMID- 11425233 TI - Submitochondrial distribution and delayed proteolysis of subunit c of the H+ transporting ATP-synthase in ovine ceroid-lipofuscinosis. AB - The neuronal ceroid-lipofuscinose (NCL) are recessively inherited lysosomal storage diseases in children and animals. The major stored protein in many of these diseases is subunit c of the mitochondrial inner membrane H+-transporting ATP-synthase. Previous studies of naturally occurring ovine ceroid-lipofuscinosis (OCL) in South Hampshire sheep showed that the genes and transcripts for subunit c were normal and inferred that this protein was expressed normally in mitochondria prior to storage in lysosomes. Accumulation in mitochondria has not been conclusively established and we have therefore used the South Hampshire model to demonstrate approximately 1.8-fold normal levels of subunit c in mitochondrial inner membranes prepared from liver. Other mitochondrial inner membrane and ATP-synthase proteins that could be detected by mass spectrometry (MS) or two-dimensional electrophoresis (2-DE) were present in normal amounts. The accumulating subunit c showed normal post-translational modification but was abnormally resistant to proteolysis. These results are consistent with the hypothesis that OCL may result from a mitochondrial disorder that affects turnover of correctly expressed subunit c, although we cannot exclude the possibility that a postmitochondrial defect delays processing of subunit c out of mitochondria. PMID- 11425234 TI - Identification of a new potential airway irritation marker, palate lung nasal epithelial clone protein, in human nasal lavage fluid with two-dimensional electrophoresis and matrix-assisted laser desorption/ionization-time of flight. AB - We have analyzed protein patterns of human nasal lavage fluid (NLF) with two dimensional gel electrophoresis (2-DE) and identified several proteins (such as transthyretin, Clara Cell protein 16, lipocalin-1, cystatin S, cystatin SN, immunoglobulin binding factor, statherin, calgranulin B, prolactin-inducible protein, and zinc-alpha2-glycoprotein) by N-terminal amino acid sequencing and matrix-assisted laser desorption/ionizationtime of flight (MALDI-TOF) mass spectrometry. To investigate whether airway irritation causes alterations in NLF 2-DE patterns, we compared epoxy workers with airway irritation (n=8) and healthy controls (n=6) before and after 2 h exposure to the epoxy chemical, dimethylbenzylamine (DMBA, 100 microg/m3) in an exposure chamber. A 25 kDa protein with pI 5.5 was found to be altered in the NLF 2-DE patterns; a trypsin digest of the 2-DE spot analyzed by MALDI-TOF and expressed sequence tags (ESTs) determined after post-source decay (PSD) identified the protein as palate lung and nasal epithelial clone (PLUNC). In controls, the levels of NLF-PLUNC were generally lower after 2 h exposure, whereas in epoxy workers, the levels were increased three- to twentyfold after exposure. The human gene sequence for PLUNC was just recently reported and so far no biofunctional data are available. Our results suggest that PLUNC is involved in the airway inflammatory response after exposure to irritants. PMID- 11425235 TI - Mapping and expression pattern analysis of key components of the major histocompatibility complex class I antigen processing and presentation pathway in a representative human renal cell carcinoma cell line. AB - Renal cell carcinoma (RCC) represent approximately 5% of all cancer deaths. At the time of presentation, over 50% of the patients have already developed locally advanced or metastatic disease with five-year survival rates of less than 20%. Although relative resistant to conventional regimens, RCC are partially susceptible to T cell-based immunotherapy. To further develop this treatment modality, two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was applied for both the mapping of the key components of the major histocompatibility complex (MHC) class I antigen processing and presentation machinery (APM) and the characterization of the constitutive and cytokine regulated protein expression profiles in a representative human RCC cell line. The latter aspect is based on the fact, that the expression level of some of the APM components can be altered in response to interferon (IFN)-gamma treatment. Total cell lysates from untreated and IFN-gamma-treated tumor cells were separated on 2-D PAGE gels using broad range immobilized pH gradient (IPG) strips. Serial Western blot analyses using sets of APM-specific antibodies were performed to target the relevant protein spots. Protein verification was mostly accomplished via peptide mass finger-printing using matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS). To date, the majority of the APM-related components have been identified and mapped. In addition, the different protein expression profiles of untreated and IFN-gamma-treated RCC cells are under investigation. PMID- 11425236 TI - Human proteome enhancement: high-recovery method and improved two-dimensional map of colostral fat globule membrane proteins. AB - The human milk fat globule membrane protein composition is still largely unknown, although it counts for 2-4% of the total milk protein content and contains several important biologically active components. The aim of this work was to create a two-dimensional electrophoresis (2-DE) map of the human milk fat globule membrane proteins, both integral and membrane-associated, and to identify and characterize as many protein components as possible. A new protocol for the solubilization and extraction of the human milk fat globule membrane proteins with a double extraction procedure is presented, and the results compared with the extraction methods reported in the literature. The proteins were separated, in the first dimension, by isoelectric focusing (IEF) in the pH range 3-10 on strips of 13 cm length and, in the second dimension, by Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) on 11.5% T homogeneous gels. A reproducible 2-DE map of integral and membrane-associated proteins was obtained and the first 23 spots, representing the major components, were identified by matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometric analysis and/or by amino acid sequencing. PMID- 11425237 TI - Inhibition of renal permeability towards albumin: a new function of apolipoproteins with possible pathogenetic relevance in focal glomerulosclerosis. AB - Focal segmental glomerulosclerosis (FSGS) is a degenerative renal disease characterized by the accumulation of extracellular matrix and lipids within the glomerular tuft. It has been proposed that an abnormal renal permeabilization towards proteins induced by a putative plasma factor is, in some way, involved in the pathogenesis of the disease. In this paper, we measured the plasma permeability activity (Palb) in several sera of patients with FSGS and found a mean activity of 0.82+/-0.03 which means a marked increase compared to a mean Palb of 0.16+/-0.03 in normal controls. Coincubation of FSGS and normal serum reduced the permeability activity within the normal range; normal serum added to the incubation medium after the glomeruli had already been exposed to the FSGS serum had no effect, suggesting the presence of inhibitory substances with a direct effect on a circulating substrate. Finally, the antipermeability activity was retained when heated to 60 degrees C but not to 100 degrees C. By serial fractionations of normal serum and reported activity measurements at each step, five natural occurring inhibitors of albumin permeabilization were purified and characterized by matrix assisted laser desorption/ionization-mass spectrometry (MALDI-MS), as components of apolipoproteins (apo) (apo E2 and E4, apo L, the high Mr apo J and a 28 kDa fragment of apo A-IV). Coincubation of each apolipoprotein with FSGS serum inhibited permeability, but only apo J and apo E2 and E4 were found to be crucial for the process. In conclusion, we have purified from normal serum five inhibitors of permeability induced by FSGS serum, all corresponding to apolipoproteins. An imbalance between permeability factors and apolipoproteins may play a pathogenetic role in FSGS. PMID- 11425238 TI - Assessing cerebrospinal fluid rhinorrhea: a two-dimensional electrophoresis approach. AB - Assessment of nasal cerebrospinal fluid (CSF) fistula commonly relies on the determination of CSF markers in an aqueous rhinorrhea, such as the beta2 transferrin immunofixation assay. While generally reliable, false positive and false negative results have been reported for most of the laboratory tests yet available. Based on the hypothesis that the simultaneous assessment of several CSF markers may yield an increased sensitivity and specificity, we used a proteomics, two-dimensional electrophoresis 2-DE based approach to study samples of nasal secretions obtained from 18 patients suspected of CSF rhinorrhea. Since CSF, nasal mucus and plasma may coexist in the nasal cavities, we first defined five specific markers for each of these biological fluids (transferrin, prostaglandin-D synthase, transthyretin, and two unknown trains of spots for CSF, immunoglobulin A (IgA) S-chain, lipocortin-1, lipocalin-1, prolactine-inducible protein and palatal lung nasal epithelium clone protein for mucus, haptoglobin alpha1/2- and beta-chains, fibrinogen alpha-, beta- and gamma-chains for plasma). Gels from the rhinorrhea patients were then compared to these 2-DE reference maps to determine the presence or absence of the defined markers, and clinical data were independently compared to the results of the 2-DE study. In all cases, the biological fluid(s) anticipated to be present in the nasal secretions based on clinical data were correctly identified by 2-DE. Moreover, an excellent correlation was found in nine patients who underwent extensive workup for suspected CSF rhinorrhea, since CSF was found by the 2-DE method in four patients in whom a CSF fistula was confirmed, whereas the test was negative in five patients in whom a CSF fistula was excluded. In the remaining patients, mucus, sometimes contamined with blood, was found to be the major component of the nasal secretions, confirming that clear mucus may mimick CSF rhinorrhea. These preliminary results suggest that a 2-DE-based multimarker approach is a valid, sensitive, and specific method to assess the presence of CSF in occult rhinorrhea. PMID- 11425239 TI - Identification of the apolipoprotein E4 isoform in cerebrospinal fluid with preparative two-dimensional electrophoresis and matrix assisted laser desorption/ionization-time of flight-mass spectrometry. AB - Apolipoprotein E (apoE) was isolated from human cerebrospinal fluid (CSF) from control individuals and patients with Alzheimer's disease (AD). The purification was performed with preparative two-dimensional electrophoresis (2-DE), involving liquid-phase isoelectric focusing (IEF) in the Rotofor cell in combination with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and electroelution in the Mini Whole Gel Eluter. ApoE was characterized by matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF MS) analysis of tryptic digests. The known change of Cys to Arg in position 112 of the apoE4 isoform was identified. This was detected in CSF from AD patients, reflecting the increased frequency of the apoE4 allele in this population. This peptide was not detected in CSF samples from healty control individuals. The use of this rapid electrophoretic separation in proteomic studies of CSF proteins provides single proteins, such as apoE, of high purity in yields sufficient for characterization by MALDI-TOF-MS. Characterization of proteins and their modifications (amino acid substitutions, glycosylation or phosphorylation) in CSF will be a useful tool in the investigation of the pathophysiology of brain disorders such as AD. PMID- 11425240 TI - Two-dimensional analysis of tear protein patterns of diabetic patients. AB - In diabetic patients, dry eye and other ocular surface diseases occur more often than in healthy subjects. The aim of this study was to analyze the tear protein patterns of patients suffering from diabetes mellitus type II (dia) and to compare them to the patterns of healthy volunteers (ctrl). Tear proteins of nonstimulated tears of 20 patients (ctrl n=10, dia n=10) were separated using two dimensional electrophoretic techniques. The protein patterns of each group were analyzed by a multivariate analysis of discriminance. Furthermore, for all spots of each gel, a 50 x 50 variables pH/Mr (molecular weight) array was generated and subsequently analyzed by a multivariate analysis of discriminance. Additionally, an artificial neural network was trained using the matrix data as input and a sensitivity analysis was performed to figure out, which spots were the most important to differentiate between the tear protein patterns. In both groups a complex staining pattern could be obtained. In diabetic patients significantly more spots were detected compared to the control group (P<0.02). The analysis of discriminance found a highly significant difference between dia and ctrl (P<0.00001). Using the matrix data, the analysis of discriminance showed a significant difference between the two groups, too (P<0.0001). The sensitivity analysis by means of the artificial neural network revealed several spots that were more expressed or more frequently present in the diabetic group. Our findings reveal that the composition of tear proteins of diabetic patients is different from that of healthy subjects. The use of the two-dimensional electrophoretic technique could give more insight into the diabetic-related changes in the tear film composition. PMID- 11425241 TI - Analysis of tear protein patterns of dry-eye patients using fluorescent staining dyes and two-dimensional quantification algorithms. AB - Tear proteins of nonstimulated tears of 29 patients (healthy subjects, n = 8; dry eye syndrome patients, n = 12; diabetic dry-eye patients, n = 9) were electrophoretically separated and stained by SYPRO Orange, followed by Coomassie blue staining. Both, the fluorescent and the Coomassie stains were subsequently analyzed by an automated two-dimensional algorithm for finding and quantification of peaks and by a discriminant analysis. Using SYPRO Orange, an average number of peaks/sample between three (at 200 ms) and 15 (at 3000 ms) could be found. In comparison, Coomassie staining resulted only in an average number of six peaks/sample. This corresponds to a sensitivity obtained at approx. 400-600 ms exposure time of SYPRO Orange stained gels. For all exposure times, the protein patterns of the three clinical groups were statistically significantly different from each other (P < 0.05). Only at 200 ms the distances between the groups decreased slightly. The Coomassie-stained gels revealed only a mid range discrimination power similar to that of 200-400 ms exposure in the fluorescing gels. The use of SYPRO Orange provides faster results than those obtained by Coomassie staining. In addition, the sensitivity of staining can be varied even in the same gel by changing the exposure time. The use of the two-dimensional algorithm allows to distinguish between the three clinical groups in accordance to earlier studies using one-dimensional densitographic raw data. Thus, the high speed of evaluation and the more sensitive results as compared to earlier studies could be a step further in the use of tear protein patterns in the diagnosis of DRY. PMID- 11425242 TI - Detection and identification of human bronchoalveolar lavage proteins using narrow-range immobilized pH gradient DryStrip and the paper bridge sample application method. AB - The use of two-dimensional gel electrophoresis as a tool for the investigation of human bronchoalveolar lavage fluid (BALF) has been hampered by technical difficulties. In the last decade attempts have been made to establish a two dimensional (2-D) protein map of BALF samples, resulting in the identification of a number of proteins present in BALF. In this study, we report an improved sample handling and separation protocol for investigation of human BALF proteins. The sample has been analyzed by employing a number of strategies, including the 'paper bridge' sample application method in combination with narrow range immobilized pH gradient (IPG) DryStrips, followed by comparison to the corresponding plasma map. Using peptide mass fingerprinting, we have identified 49 proteins in the narrow pH range 4.5-5.2 from an individual healthy BALF sample. Furthermore, we identified 17 BALF proteins, not detected in plasma. Twelve of these proteins have, to our knowledge, not previously been described in the BALF 2-D map. The mapping of BALF proteins with inclusion of those at low concentration increases the possibility to subsequently screen patient material for disease markers. PMID- 11425243 TI - Differential diagnosis of prostate cancer and benign prostate hyperplasia using two-dimensional electrophoresis. AB - Prostate specific antigen (PSA) is a protease which is characteristic of the prostate. It is widely used as a serum marker for the early diagnosis of prostate cancer (PCa). Nevertheless, for concentrations between 4 and 10 ng/mL, PSA does not enable PCa to be distinguished from benign diseases, such as benign prostate hyperplasia (BPH). In sera, the use of a ratio between free PSA (PSA uncomplexed with protease inhibitor) and total PSA (free PSA and PSA bound to alpha-1 anti chymotrypsin) enables the "gray zone" to be reduced, but an important proportion of patients are still wrongly classed. Using two-dimensional electrophoresis, we demonstrated using 52 PCa and 40 BPH well-documented clinical cases that BPH sera show a significantly greater percentage of low-molecular-weight free PSA elements (IwPSA) than PCa sera. In our study, the use of a ratio between IwPSA and standard free PSA enables the correct diagnosis of 100% of PCa and 82.5% of BPH cases as against when 73.1% and 42.5% respectively were correctly diagnozed using the total PSA and the free/total PSA ratio. This important finding may be related to differences in the mechanism secreting PSA from the prostate into the bloodstream. We have shown how a tissue marker may be turned into a powerful tumor marker by events probably unrelated to its expression. PMID- 11425244 TI - Two-dimensional database of a Burkitt lymphoma cell line (DG 75) proteins: protein pattern changes following treatment with 5'-azycytidine. AB - Hypermethylation is an important mechanism for repression of tumor gene suppressor in cancer. The drug 5'-azacytidine (AZC) has been used as demethylating agent to induce the expression of previously silencing genes. In the present work, we attempted to determine, using proteomics, the changes in protein expression profiles following a treatment of an Epstein Barr virus (EBV) negative Burkitt lymphoma (BL) cell line DG 75. The effects of the treatment in terms of cell viability and growth were first examined. The following observations were made: AZC treatment led to (i) a decrease in cell growth with an arrest of the cell at G0/G1 phase of the cell cycle, (ii) the expression of p16, a tumor-suppressor gene whose expression was dependent on its promoter demethylation. Proteomic study evidenced that AZC treatment affected protein expression in two different ways. Twenty-one polypeptides were down-expressed, while 14 showed an increased expression. Some of the upregulated proteins appeared related to the energy metabolism, to organization of cytoskeletal structures, and to cell viability and protein synthesis. We also established a reference map for proteins in DG 75 cell line, comprising 74 different polypeptides corresponding to 67 proteins. This map will be accessible via Internet as a resource for proteome analyses of B-cells. Taken together, the results presented here highlight new insights into lymphoma cell gene regulations following a treatment of lymphoma cells with AZC and illustrate a use of proteomics to evidence the direct and indirect effects of a drug and the pathways it possibly regulates. PMID- 11425245 TI - The changing face of dental education: the impact of PBL. AB - The past decade has seen increasing demands for reform of dental education that would produce a graduate better equipped to work in the rapidly changing world of the twenty-first century. Among the most notable curriculum changes implemented in dental schools is a move toward Problem-Based Learning (PBL). PBL, in some form, has been a feature of medical education for several decades, but has only recently been introduced into dental schools. This paper discusses the rationale for the introduction of a PBL pedagogy into dental education, the modalities of PBL being introduced, and the implications of the introduction of PBL into dental schools. Matters related to implementation, faculty development, admissions, and assessment are addressed. Observations derived from a parallel-track dental PBL curriculum at the University of Southern California (USC) are presented and discussed. This program conforms to the Barrows (1998) concept of "authentic PBL" in that the program has no scheduled lectures and maintains a PBL pedagogy for all four years of the curriculum. The USC dental students working in the PBL curriculum have attained a high level of achievement on U.S. National Dental Boards (Part I) examinations, significantly superior to their peers working in a traditional lecture-based curriculum. PMID- 11425246 TI - Assessing outcomes of curricular change: a view from program graduates. AB - Graduates of dental programs constitute a stakeholder group that is able to provide unique information concerning the effectiveness of the dental curriculum in preparing them for dental careers. Following the implementation of planned curricular changes, graduates of the former and the new curricula were surveyed. Results indicate that, while both groups perceive themselves to have been adequately prepared by their D.D.S. program for practice, graduates of the new curriculum indicated a higher level of preparedness in several areas, especially regarding selecting, prescribing and administering pharmacotherapeutic agents. In addition, the results suggest that the change in the first two years from an emphasis on basic sciences and preclinical laboratory work to an early introduction to live dental patients and the integration of foundation sciences and clinical courses were endorsed. The evidence gathered from this alumni survey indicates that the reforms implemented in the dental curriculum were appropriate. Areas requiring further investigation are also discussed. PMID- 11425247 TI - The EXCEL Program: strengthening diversity. AB - The Boston University Henry M. Goldman School of Dental Medicine (BUSDM) initiated a program in the summer of 1993 to strengthen diversity in the entering class of first-year students. The Experiential Center for Excellence in Learning (EXCEL) Program is a voluntary, one-month-long prematriculation experience that combines didactic, laboratory, study skills, and social activities to prepare participants to transition into the rigorous first-year curriculum. From 1996 to 2000, ninety students participated in EXCEL. The two primary reasons cited for participating were to become familiar with the school, faculty, and classmates and to strengthen basic science background. Participants' ages ranged from twenty to over forty. Fifty-nine percent of participants had been out of college for more than one year; 10 percent had been out of school for three years or more. Thirty percent listed nontraditional predental school majors. Fifty-six percent listed a country other than the United States as country of birth. Of those completing an exit survey, 96 percent reported that EXCEL strengthened their decision to study dentistry, and 97 percent would recommend that future entering BUSDM students participate in EXCEL. The EXCEL Program may serve as a model for increasing diversity in U.S. dental school enrollment. PMID- 11425248 TI - Occupational exposures to blood in a dental teaching environment: results of a ten-year surveillance study. AB - Evaluation of occupational exposures can assist with practice modifications, redesign of equipment, and targeted educational efforts. The data presented in this report has been collected as part of a ten-year surveillance program of occupational exposures to blood or other potentially infectious materials in a large dental teaching institution. From 1987 to 1997, a total of 504 percutaneous/non-intact skin and mucous membrane exposures were documented. Of these, 494 (98 percent) were percutaneous, and 10 (2 percent) were mucosal, each involving a splash to the eye of the dental care worker (DCW). Among the 504 exposures, 414 (82.1 percent) occurred among dental students, 60 (11.9 percent) among staff, and 30 (6 percent) among faculty. One hundred ninety-one (37.9 percent) exposures were superficial (no bleeding), 260 (51.6 percent) were moderate (some bleeding), and 53 (10.5 percent) were deep (heavy bleeding). Regarding the circumstances of exposure, 279 (54.5 percent) of the injuries occurred post-operatively (after the use of the device), and most were related to instrument clean-up; 210 (41.0 percent) occurred intra-operatively (during the use of the device); and 23 (4.5 percent) occurred when a DCW collided with a sharp object in the dental operatory (eight cases involved more than one circumstance). The overall exposure rate for the college was 2.46+/-0.11 SD per 10,000 patient visits. The average rate for the student population was 4.02+/ 0.20 SD per 100 person-years, with the highest rates being observed among junior year students. The observed rates of occupational exposures to blood and body fluids in this report are consistent with published reports from several other educational settings. Dental teaching institutions are faced with the unique challenge of protecting the student and patient populations against bloodborne infections. Educational efforts must go beyond mere teaching of universal precautions and should include the introduction of safer products and clinical procedures that can minimize the risks associated with the hands-on aspects of the students' learning process. PMID- 11425249 TI - Variation, certainty, evidence, and change in dental education: employing evidence-based dentistry in dental education. AB - Variation in health care, and more particularly in dental care, was recently chronicled in a Readers Digest investigative report. The conclusions of this report are consistent with sound scientific studies conducted in various areas of health care, including dental care, which demonstrate substantial variation in the care provided to patients. This variation in care parallels the certainty with which clinicians and faculty members often articulate strongly held, but very different opinions. Using a case-based dental scenario, we present systematic evidence-based methods for accessing dental health care information, evaluating this information for validity and importance, and using this information to make informed curricular and clinical decisions. We also discuss barriers inhibiting these systematic approaches to evidence-based clinical decision making and methods for effectively promoting behavior change in health care professionals. PMID- 11425250 TI - Tissue engineering's impact on dentistry. AB - Tissue engineering is a novel and exciting field that aims to re-create functional, healthy tissues and organs in order to replace diseased, dying, or dead tissues. The field has developed due to the inadequate supply of organs and tissues for patients requiring organ and tissue replacement. The following review first describes three major tissue engineering strategies. Although similar in their objectives, these strategies each maintain a unique component. Next, several examples of preclinical and clinical progress engineering oral maxillofacial tissues are presented. Each of these examples highlights specific tissue engineering applications to different tissues of the oral-maxillofacial apparatus. Finally, practical implications are addressed as well as challenges that must be met in order for tissue engineering to reach its full potential. PMID- 11425251 TI - The human genome, implications for oral health and diseases, and dental education. AB - We are living in an extraordinary time in human history punctuated by the convergence of major scientific and technological progress in the physical, chemical, and biological ways of knowing. Equally extraordinary are the sparkling intellectual developments at the interface between fields of study. One major example of an emerging influence on the future of oral health education is at the interface between the human genome, information technology, and biotechnology with miniaturizations (nanotechnology), suggesting new oral health professional competencies for a new century. A great deal has recently been learned from human and non-human genomics. Genome databases are being "mined" to prompt hypothesis driven "postgenomic" or functional genomic science in microbial models such as Candida albicans related to oral candidiasis and in human genomics related to biological processes found in craniofacial, oral, and dental diseases and disorders. This growing body of knowledge is already providing the gene content of many oral microbial and human genomes and the knowledge of genetic variants or polymorphisms related to disease, disease progression, and disease response to therapeutics (pharmacogenomics). The knowledge base from human and non-human genomics, functional genomics, biotechnology, and associated information technologies is serving to revolutionize oral health promotion, risk assessment using biomarkers and disease prevention, diagnostics, treatments, and the full range of therapeutics for craniofacial, oral, and dental diseases and disorders. Education, training, and research opportunities are already transforming the curriculum and pedagogy for undergraduate science majors, predoctoral health professional programs, residency and specialty programs, and graduate programs within the health professions. In the words of Bob Dylan, "the times they are a changing." PMID- 11425252 TI - Trends in allied dental education: an analysis of the past and a look to the future. AB - Allied dental healthcare providers have been an integral part of the dental team since the turn of the 19th century. Like dental education, allied dental education's history includes a transition from apprenticeships and proprietary school settings to dental schools and community and technical colleges. There are currently 258 dental assisting programs, 255 dental hygiene programs, and 28 dental laboratory technology programs according to the American Dental Association's Commission on Dental Accreditation. First-year enrollment increased 9.5 percent in dental hygiene education from 1994/95 to 1998/99, while enrollment in dental assisting programs declined 7 percent and declined 31 percent in dental laboratory technology programs during the same period. Program capacity exceeds enrollment in all three areas of allied dental education. Challenges facing allied dental education include addressing the dental practicing community's perception of a shortage of dental assistants and dental hygienists and increasing pressure for career tracks that do not require education in ADA Commission on Dental Accreditation accredited programs. The allied dental workforce may also be called upon for innovative approaches to improve access to oral health care and reduce oral health care disparities. In addition, allied dental education programs may face challenges in recruiting faculty with the desired academic credentials. ADEA is currently pursuing initiatives in these and other areas to address the current and emerging needs of allied dental education. PMID- 11425253 TI - Foot-and-mouth disease: continuing concern about the 'Settle cluster'. PMID- 11425254 TI - Aetiology of clinical mastitis in six Somerset dairy herds. AB - Clinical mastitis was monitored in six Somerset dairy herds for one year. The herds all had three-month geometric mean bulk milk somatic cell counts of less than 250,000 cells/ml. Escherichia coli was the predominant pathogen isolated on all the farms and in all months of the year. Environmental pathogens accounted for 61.4 per cent of all cases of clinical mastitis and for 79.3 per cent of the mastitis cases in which an aetiological agent was identified. The mean annual incidence was 41.6 cases per 100 cows (range 14 to 75). Affected cows suffered a mean of 1.5 cases and 16.4 per cent of quarters suffered at least one repeat case. Mastitis due to E. coli was more severe than mastitis due to other causes and it tended to be more severe in early lactation and during the housing period. Mastitis was significantly more severe (grades 2 and 3) in the herd with the lowest bulk milk somatic cell count and in the herd which was kept indoors throughout the year than in the other four herds. Mastitis was fatal in 2.2 per cent of cases and resulted in the death of 0.6 per cent of the lactating cows. PMID- 11425255 TI - Diagnostic investigation of chronic porcine reproductive and respiratory syndrome virus in a breeding herd of pigs. AB - Forty-five sows and 15 boars were selected at random from a breeding herd known to be chronically infected with porcine reproductive and respiratory syndrome virus (PRRSV) and lymphoid, immune-privileged, and non-lymphoid/non-immune privileged tissues were tested for the presence of the virus by PCR, virus isolation, and immunohistochemistry. The virus was isolated from the lateral retropharyngeal lymph node of one sow; the isolate was nucleic acid sequenced and determined to be of field origin, and it was inoculated into two PRRSV-naive pregnant sows (A and B) at 95 days of gestation. They were necropsied 14 days later and samples of maternal and fetal tissue and blood samples were collected. Sow A had 10 fresh, six partially autolysed, and two mummified fetuses, and sow B had six fresh and viable fetuses. Viral nucleic acid was detected by PCR in tissue pools from each sow and also from pooled fetal tissues, and the virus was isolated from fetal pools from sow A. PMID- 11425256 TI - Protection of puppies against canine herpesvirus by vaccination of the dams. AB - Six bitches free of canine herpesvirus 1 (CHV-1) were vaccinated against the virus; a first injection was given 10 days after the presumed date of mating and a second six weeks later. Six similar bitches were left unvaccinated as controls, and all the pups were challenged oronasally with a virulent strain of CHV-1 at three days of age. All the vaccinated bitches seroconverted and had high antibody titres when the puppies were challenged, but the control bitches remained seronegative. In the control group, 62 per cent (18 of 29) of the pups died of CHV-1-induced disease; most of them showed typical clinical signs and macroscopic lesions, and CHV-1 infection was confirmed by the isolation of the virus or by PCR. None of the puppies in the vaccinated group died of CHV-1 infection. The efficacy of the vaccine was confirmed in CHV-1-positive breeding units. The rate of pregnancy tended to be higher in vaccinated bitches and the mortality of pups before weaning was significantly reduced in the litters born to vaccinated bitches. PMID- 11425257 TI - Evaluation of imidacloprid for the treatment and prevention of cat flea (Ctenocephalides felis felis) infestations on rabbits. PMID- 11425258 TI - Traumatic separation of the auricular and annular cartilages in two cats. PMID- 11425259 TI - FMD control strategies. PMID- 11425260 TI - FMD control strategies. PMID- 11425261 TI - Dispensing of medicines. PMID- 11425262 TI - Burring cheek teeth in rabbits. PMID- 11425263 TI - Characteristics of malignant tumors in young people, with particular emphasis on carcinomas and sarcomas. AB - The aims of the study are to investigate the characteristic patterns related to different aged subgroups of young Chinese patients with malignant tumors. The subjects were young Chinese patients (defined as age > 1 and < 40 years) who underwent autopsy for malignant tumors in a teaching hospital in Hong Kong from 1970 to 1999. They were divided into four age groups: 1 to 9 years, 10 to 19 years, 20 to 29 years, and 30 to 39 years. The malignant tumors were classified into subgroups according to the pathological classification. The clinicopathologic features of patients with carcinomas and sarcomas were reviewed in depth. Four hundred sixty-three (22%) of 2,080 young patients had malignant tumors detected at autopsies. The male-to-female ratio was 1.5. The two most common malignant tumors were malignant lymphoid tumors and carcinomas, accounting for 49.9% and 29.8%, respectively, of all malignant tumors. In children (ages 1-9 years), 78.9% of malignant tumors found were malignant lymphoid tumors and neuroendocrine tumors. In teenagers (ages 10-19 years), malignant lymphoid tumors were the most important group of malignant tumor, accounting for 69% of malignant tumors in this age group. Carcinomas and malignant lymphoid tumors were seen in more than 80% of malignancies in young adults (ages 20-39). Overall, carcinomas and sarcomas were noted in 29.8% and 4.1%, respectively, of young patients with malignant tumors. The common primary sites of carcinomas were liver, stomach, lung, and nasopharynx. Sarcomas were more common in females (11 females, seven males), and rhabdomyosarcoma was the most common sarcoma found. Compared with other malignant tumors, carcinomas were less often diagnosed before death. In conclusion, malignant tumor is a common cause of death in young patients. Different types of malignant tumors were seen in various groups of young patients. PMID- 11425264 TI - Differential expression of metallopanstimulin/S27 ribosomal protein in hepatic regeneration and neoplasia. AB - We have previously shown that human metallopanstimulin-1 (MPS-1) is a ubiquitous 9.4-kd multifunctional ribosomal S27/nuclear "zinc finger" protein that is expressed at high levels in a wide variety of actively proliferating cells and tumor tissues. In this study, we examined the expression of MPS-1 in chronic hepatitis, cirrhosis, and hepatocellular carcinoma. Tissue samples were obtained at the time of tumor resection, needle biopsy, or liver transplantation. MPS- 1 was studied by immunohistochemistry by use of specific antibodies to the N terminus of MPS-1 in a biotin/streptavidin-amplified system. In chronic hepatitis, hepatocytes had very weak MPS-1 immunostaining. In contrast, hepatocytes in regenerating cirrhotic nodules stained strongly for MPS-1. In well differentiated hepatocellular carcinoma, MPS-1 presence was intense at the periphery of the malignant nodule. In poorly differentiated hepatocellular carcinoma, MPS-1 presence was notably intense in malignant hepatocytes invading the septal tissues, in close contact with neovascular structures. These results suggest that MPS-1 may be involved in both progression toward malignancy in regenerating cirrhotic nodules and in subsequent steps of hepatocarcinogenesis. PMID- 11425265 TI - Allelotype of the adenoma-carcinoma sequence of the stomach. AB - In order to identify the significant allelic loss involving the gastric adenoma carcinoma sequence, we used 49 genome-wide microsatellite markers in an allelotype study of 30 cases of stomach resections that harbored both adenomas and carcinomas. Frequent loss of heterozygosity was demonstrated on 12q (53.3%), 2p (50.0%), and 18p (50.0%) in adenomas and on 8q (80.0%), 2p (70.0%), 18p (66.7%), and 17p (61.9%) in carcinomas. Significant difference in the loss of heterozygosity rate between the adenoma and the carcinoma was noted on 17p. Our results suggested that the critical target of loss of heterozygosity in gastric adenomacarcinoma sequences may be the p53 gene on 17p. PMID- 11425266 TI - Plasma micronutrient antioxidant in cancer patients. AB - The distribution of breast, colon, gastric, thyroid, oral, rectal, pancreatic and renal cancers were determined in 71 Kuwaitis, 45 other Arabs, and 26 Indians. Plasma levels of micronutrient antioxidants, retinol, alpha-tocopherol, lycopene, and beta-carotene were measured in the groups and in 90 matched controls for comparison. Cholesterol was measured to determine its association with the micronutrient antioxidants. Pancreatic cancer occurred exclusively in Kuwaitis, while breast and colon cancers were disproportionately higher in Kuwaitis than in the other groups. Micronutrient antioxidant levels were similar in the groups, except for higher lycopene levels in Kuwaitis. In most instances, the micronutrient antioxidants, except beta-carotene, decreased significantly in levels in patients than in controls. Low levels of retinol, lycopene, and beta carotene were strongly associated with pancreatic cancer. Compared to controls, significantly increased levels of beta-carotene occurred in breast, colon, thyroid, and renal cancers; increased lycopene occurred in oral cancer, and increased alpha-tocopherol occurred in pancreatic cancer. Alpha-tocopherol strongly correlated with cholesterol. Generally, changes in alpha-tocopherol/ cholesterol ratios mimicked those of alpha-tocopherol levels. Micronutrient antioxidant levels were significantly lower in male patients than female patients. Age showed a negative but statistically insignificant relationship with micronutrient antioxidants. Lycopene strongly correlated with alpha-carotene and alpha-tocopherol with retinol. Among the patients, all micronutrient antioxidants except retinol decreased significantly in levels in smokers than nonsmokers, suggesting susceptibility to cigarette smoke oxidative stress. We conclude that micronutrient antioxidant depletions and altered associations may imply tumor utilization or antioxidant burden in oxidative stress or both. Furthermore, the incidence of pancreatic, colon and breast cancers among Kuwaitis warrants further study. PMID- 11425267 TI - Feeding thiol-containing compounds, derived from vegetables, fails to inhibit N methylnitrosourea-induced mammary tumorigenesis. AB - Various thiol-containing compounds have been shown to inhibit chemically-induced tumors in animal models. Two thiol-containing compounds derived from vegetables, namely 1,2 dithiol-3-thione (DTT) and S-methylmethane thiolsulfonate (MMTS), were tested for their chemopreventive activity in the N-methylnitrosourea (NMU) induced rat mammary tumor model. Each compound was incorporated into the grain based Teklad 7001 diet and fed to the rats one week prior to initiation with NMU until termination 18 weeks post NMU. DTT was fed at 166 and 500 ppm and MMTS at 200 and 800 ppm. Neither compound exerted a significant inhibitory effect on any index of tumor development including incidence, total tumor, tumor multiplicity, volume or latency. Serum levels of DTT assessed at termination in the 500 ppm DTT group ranged from 10-30 microg/ml. MMTS was undetectable in serum from either MMTS-fed group. The results of this study, using the direct acting carcinogen, NMU, suggest that the chemopreventive effect of thiol-containing compounds may be confined to animal models using carcinogens that require host activation. PMID- 11425268 TI - Loss of heterozygosity and allelic imbalance in apocrine adenosis of the breast. AB - Recently, there have been studies suggesting that apocrine adenosis of the breast is a putative precancerous lesion, despite the generally held view that apocrine adenosis is benign. Because apocrine adenosis is almost always present as a small area or areas, it cannot be easily studied by conventional methods. In this study, areas of apocrine adenosis were microdissected from archival paraffin embedded tissue to examine loss of heterozygosity and allelic imbalance compared with normal breast tissue epithelium from the same patients. Seventeen cases of apocrine adenosis, four associated with carcinoma, were analyzed using polymorphic microsatellite markers and polymerase chain reaction for loss of heterozygosity/allelic imbalance at eight loci that were reported to show allele loss or imbalance in invasive and in situ breast cancer. Loss of heterozygosity/allelic imbalance was detected in six of 17 cases of apocrine adenosis; three of 12 (25%) informative cases at 1p (MYCL1), two of seven (28.6%) at 11q (INT2), one of three (33.3%) at 13q (D13S267), two of 12 (16.7%) at 16q (D16S539), and two of 10 (20%) at 17q (D17S250). Neither loss of heterozygosity nor allelic imbalance has been identified at 1p (D1S252), 17p (TP53), or 17p (D17S513). In two of the four cases associated with carcinoma, loss of heterozygosity/allelic imbalance was seen in the same allele as in the synchronous carcinoma. These results suggest that molecular alterations, such as loss of heterozygosity and allelic imbalance, identified in apocrine adenosis may constitute an early event in the pathogenesis of breast cancer; reinforcing the possibility of apocrine adenosis being a putative precancerous lesion. PMID- 11425269 TI - Comparative analysis of two-dimensional protein patterns in malignant and normal human breast tissue. AB - Malignant and normal human breast tissue were compared by evaluating two dimensional polyacrylamide gel electrophoresis (2D-PAGE) maps of frozen tissue samples. Image analyzing software was used to scan and process 34 gels. Eight (8/34) of these gels (4 malignant breast tumor samples, 4 normal tissue samples) were selected on the basis of gel and image quality to build a database to identify and measure the expression of a previously unidentified proteome. Growth factor receptor proteins (GFRs), including ERBB2 (HER2) and ERBB3 (HER3), were expressed in the malignant tissue samples. Growth factor receptor proteins were not expressed in the normal tissue. Also, expression of PS2-protein (pS2) was detected in neither malignant nor normal tissue. In benign breast samples a higher intensity of protein expression could be observed for maspin, desmoglein 3 and keratin 8 than in malignant samples. Other proteins expressed in malignant breast tissue include mitogen-activated protein kinase 3 (MK03), heat shock protein 27 kDa (HS27), growth factor receptor-bound protein (GRB2), cathepsin D, G1/S specific cyclin E1 (CGEI), glucose transporter type 5 (GTR5), and a number of as yet unidentified proteins. PMID- 11425270 TI - Risk factors for benign breast biopsies: a nested case-control study in the Alberta breast screening program. AB - A nested case-control study was conducted to identify risk factors for benign breast biopsies in 382 cases (women with a benign biopsy result) and 399 controls (women who had not undergone a biopsy) who were sampled from the Alberta breast cancer screening program. The breast biopsy specimens of the cases were reviewed by a panel of pathologists, and percent fibroglandular tissue density was assessed. The multivariable odds ratios for the risk of open benign breast biopsy associated with current cigarette smoking was 2.04 (95% CI 1.32-3.13), for ever regular smoking was 1.61 (1.20-2.16), and for passive smoking was 1.41 (0.99 2.02). A risk reduction was found for ever alcohol consumption (0.61 [0.44 0.85]). Some risk reductions were found when the highest and the lowest quintiles of total aerobic recreational activity were compared (0.71 [0.42-1.20]), stair climbing (0.61 [0.37-1.01]) and walking pace (0.13 [0.02-0.741). Lifestyle risk factors may be implicated in the continuum between detection of an abnormality on a screening mammogram and a breast biopsy specimen. By considering these risk factors, breast screening programs may be better able to identify those women who require a breast biopsy and reduce the number of benign breast biopsies. PMID- 11425271 TI - Alterations of the FHIT gene in breast cancer: association with tumor progression and patient survival. AB - Our previous results on breast tumors show that LOH (loss of heterozygosity) at the FHIT locus is associated with reduced Fhit protein expression. We have also shown that LOH at this locus is significantly higher in tumors from patients carrying the BRCA2 999de15 mutation than in tumors without this mutation, presumably because of lack of DNA repair. Here, our aim was to determine the relationship of FHIT LOH with breast tumor progression. Five microsatellite markers located within the FHIT gene were typed in 239 breast tumors and corresponding normal tissue, and the LOH results were compared with clinicopathologic factors and LOH at other chromosome regions. LOH at FHIT is associated with estrogen- and progesterone-negative breast tumors, high S-phase fraction, reduced patient survival, and LOH at chromosome regions 6q, 7q, 8p, 9p, 11p, 11q, 13q, 16q, 17p, 17q, 18q, and 20q. A multivariate analysis shows that LOH at FHIT results in a 60% increased relative risk of dying. We conclude that the loss of FHIT results in growth advantage of breast tumor cells, is associated with unstable genome, and may be of prognostic value. PMID- 11425272 TI - Preoperative serum levels of sialyl Lewis(a), sialyl Lewis(x), and sialyl Tn antigens as prognostic markers after curative resection for colorectal cancer. AB - In this study, we examined the preoperative serum levels of sialyl Lewisa, sialyl LewisX, sialyl Tn, and carcinoembryonic antigen in 243 colorectal cancer patients in order to clarify the role of these antigens as prognostic factors after curative surgery. The patients were divided into two groups: low and high antigen groups (lower and higher than a selected diagnostic-based cut-off value). Patients with high serum levels of sialyl Lewisa and carcinoembryonic antigen had shorter disease-free intervals than those with low serum levels of the respective antigen, although sialyl Lewisx and sialyl Tn showed no significant differences. Multivariate analysis revealed that three independent prognostic variables, including depth of tumor invasion, lymph node metastasis, and serum sialyl Lewisa level, did prove to have value in predicting disease-free interval. In conclusion, among the four antigens examined in this study, the preoperative serum level of sialyl Lewisa is the only independent prognostic variable for recurrence after curative resection of colorectal cancer. PMID- 11425273 TI - Colorectal cancer screening by primary care physicians in two medical care organizations. AB - The objectives of this study were to (1) evaluate self-reported practices of colorectal cancer (CRC) screening among primary care physicians (PCPs) in medical organizations and (2) identify factors associated with screening performance. We surveyed a census sample of 204 PCPs from two medical organizations in Houston, TX. Outcomes were PCPs' self-reports of screening with fecal occult blood test (FOBT) and/or flexible sigmoidoscopy (FS).2-7 Independent variables included physician demographics, perception of screening efficacy, level of agreement with screening guidelines, and perceptions of screening barriers. Variables were compared using Student's t-tests, Chi-square, and Fisher's exact tests. Regression was used to examine factors associated with PCPs' reports of screening. Our response rate was 56% (n = 115). Many PCPs reported recommending and/or performing/ordering screening with both FOBT (82%) and FS (87%). They more often reported believing that FS was "very effective" for reducing mortality than was FOBT (76% vs. 37%). Physicians perceived that barriers to patient compliance with CRC screening were stronger than barriers to making screening recommendations. Significant test-specific differences in reported barriers to screening were found. Results revealed high awareness and self-reported practice of CRC screening in this study population. Further examination of how barriers influence CRC screening practices by PCPs in medical organizations will be important for improving population screening rates. PMID- 11425274 TI - Suppression of tubulin tyrosine ligase activity through reversible phosphorylation: a mechanism for inhibition of alpha-tubulin tyrosinylation. AB - The tubulin tyrosinylation/detyrosinylation cycle is a well-established posttranslational modification, which is carried out by two enzymes: tubulin tyrosine ligase (TTL) and tubulin tyrosine carboxypeptidase (TTCP). In this paper, I present evidence suggesting that the cycle itself is under the hierarchical control of reversible phosphorylation and that proteinkinase C (PKC) mediated phosphorylation of TTL inhibits its activity, thereby preventing tubulin tyrosinylation. Phosphorylation of TTL is postulated to occur in its presumed Mg(++)-ATP binding fold, leading to inhibition of Mg(++)/ATP binding and TTL mediated catalysis. The implications of such control are also discussed. Copyright 2001 Harcourt Publishers Ltd. PMID- 11425275 TI - A possible explanation for the frequent concomitance of arterial hypertension and multiple renal arteries. AB - In more than 20% of subjects, at least one kidney is found to be supplied by more than one artery arising from the aorta. This aberrant renovascular anatomy has been reported in the literature to occur in up to 80% of patients who suffer from essential hypertension. Predominant numbers of the so-called 'accessory' vessels are longer and narrower than the segmental arteries arising in the main renal artery. As a result (in accordance with Poiseuille's law of fluid flow), the renal segments supplied by these 'accessory' vessels may have lower levels of blood pressure than the remainder of the parenchyma, thereby increasing the renin secretion. This hypothesis could be significant in terms of finding a causal treatment for a disorder induced by such a mechanism. We first review the literature in which the frequency of these vascular anomalies in normotensive and hypertensive patients is described, and then advance a hypothesis explaining the frequent incidence of essential hypertension in these subjects, as well as the ramifications of this phenomenon. PMID- 11425276 TI - A mathematical model of internal time processing in temporal lobe epilepsy. AB - Based on known relationships between epileptic and nonepileptic cortical cerebral blood flow, electrocorticographic factors and epileptogenicity, a mathematical model for internal time processing is derived. The model suggests that the human brain has mechanisms for internal processing of real, reverse and imaginary time. PMID- 11425277 TI - Current prospects for controlling cancer growth with non-cytotoxic agents- nutrients, phytochemicals, herbal extracts, and available drugs. AB - In animal or cell culture studies, the growth and spread of cancer can be slowed by many nutrients, food factors, herbal extracts, and well-tolerated, available drugs that are still rarely used in the clinical management of cancer, in part because they seem unlikely to constitute definitive therapies in themselves. However, it is reasonable to expect that mechanistically complementary combinations of these measures could have a worthwhile impact on survival times and, when used as adjuvants, could improve the cure rates achievable with standard therapies. The therapeutic options available in this regard include measures that: down-regulate serum free IGF-I; suppress the synthesis of mevalonic acid and/or certain derivatives thereof; modulate arachidonate metabolism by inhibiting 5-lipoxygenase, 12-lipoxygenase, or COX-2; antagonize the activation of AP-1 transcription factors; promote the activation of PPAR gamma transcription factors; and that suppress angiogenesis by additional mechanisms. Many of these measures appear suitable for use in cancer prevention. PMID- 11425278 TI - Many Gulf War illnesses may be autoimmune disorders caused by the chemical and biological stressors pyridostigmine bromide, and adrenaline. AB - Gulf War-related illnesses are mostly common ailments, but with incidence rates that exceed those expected in the population of Gulf War veterans. These illnesses may be the result of combinations of chemical and physiological stressors which may have caused acute cellular effects sufficient to initiate processes of autoimmunity to various organs, tissues or types of cells. Two main suspects in the Gulf War cluster of illnesses are the 'Nerve Gas Pill' (pyridostigmine bromide, PB, NAPS) and stress. One component of stress, beta adrenergic load, potentiates the toxicity of PB. While similar types of chemical and physiological stressors are present in the general population, the Gulf War veteran population received these stressors in a short time, with greater intensity, and at a higher percentage exposure than normal for the general population. This may be an opportunity to learn the cause, how to prevent, and, possibly, how to treat these ailments in Gulf War veterans and in the general population. PMID- 11425279 TI - The importance of cultural inheritance. AB - Cultural inheritance refers to the storage and transmission of information by communication, imitation, teaching and learning. It is transmitted by the brain rather than by genes. However, it does have a genetic basis, the genes involved determining the structure of the brain. Cultural inheritance is considered to be the latest stage in the evolution of heredity. It is thought to have evolved by epigenetic mechanisms from genetic inheritance. This article proposes that cultural inheritance underlies normal behaviour and mental disorders. PMID- 11425280 TI - A pendular mechanism for maintenance of the immune memory. AB - A possible mechanism for maintaining immune memory, based on idiotypic-anti idiotypic interactions, is described. It is proposed that pendular dynamic swings in the levels of idiotypic antibodies (Ab1) and anti-idiotypic Ab2 may underlay immune memory. In the terms of the advanced concept, Ab dynamics in the maternal body might also play a significant role in education of the neonatal immune system. PMID- 11425281 TI - The multifaceted and widespread pathology of magnesium deficiency. AB - Even though Mg is by far the least abundant serum electrolyte, it is extremely important for the metabolism of Ca, K, P, Zn, Cu, Fe, Na, Pb, Cd, HCl, acetylcholine, and nitric oxide (NO), for many enzymes, for the intracellular homeostasis and for activation of thiamine and therefore, for a very wide gamut of crucial body functions. Unfortunately, Mg absorption and elimination depend on a very large number of variables, at least one of which often goes awry, leading to a Mg deficiency that can present with many signs and symptoms. Mg absorption requires plenty of Mg in the diet, Se, parathyroid hormone (PTH) and vitamins B6 and D. Furthermore, it is hindered by excess fat. On the other hand, Mg levels are decreased by excess ethanol, salt, phosphoric acid (sodas) and coffee intake, by profuse sweating, by intense, prolonged stress, by excessive menstruation and vaginal flux, by diuretics and other drugs and by certain parasites (pinworms). The very small probability that all the variables affecting Mg levels will behave favorably, results in a high probability of a gradually intensifying Mg deficiency. It is highly regrettable that the deficiency of such an inexpensive, low-toxicity nutrient result in diseases that cause incalculable suffering and expense throughout the world. The range of pathologies associated with Mg deficiency is staggering: hypertension (cardiovascular disease, kidney and liver damage, etc.), peroxynitrite damage (migraine, multiple sclerosis, glaucoma, Alzheimer's disease, etc.), recurrent bacterial infection due to low levels of nitric oxide in the cavities (sinuses, vagina, middle ear, lungs, throat, etc.), fungal infections due to a depressed immune system, thiamine deactivation (low gastric acid, behavioral disorders, etc.), premenstrual syndrome, Ca deficiency (osteoporosis, hypertension, mood swings, etc.), tooth cavities, hearing loss, diabetes type II, cramps, muscle weakness, impotence (lack of NO), aggression (lack of NO), fibromas, K deficiency (arrhythmia, hypertension, some forms of cancer), Fe accumulation, etc. Finally, because there are so many variables involved in the Mg metabolism, evaluating the effect of Mg in many diseases has frustrated many researchers who have simply tried supplementation with Mg, without undertaking the task of ensuring its absorption and preventing excessive elimination, rendering the study of Mg deficiency much more difficult than for most other nutrients. PMID- 11425282 TI - Is Parkinson's disease the heterozygote form of Wilson's disease: PD = 1/2 WD? AB - Wilson's disease (WD) patients often present with Parkinson's disease (PD). Furthermore, most patients with PD have reduced ceruloplasmin, a characteristic of Wilson's disease. WD is an autosomal recessive disease (requires two faulty copies of a gene to produce a homozygote individual) that afflicts 1 in 1000 people. However, the number of people with one faulty copy (heterozygotes) is much larger, probably about 2% of the population. I hypothesize that the large number of heterozygotes for WD are at greatly increased risk for idiopathic PD, because these people accumulate free copper in the basal ganglia at a slower rate than homozygotes, which accounts for the fact that PD usually develops after 40 years of age. In WD, a ceruloplasmin deficiency results in accumulation of free Cu in the liver, brain, kidneys, etc. The excess Cu results in impaired Zn absorption, which would account for the low levels of Zn in the brains of PD patients. Moreover, the high levels of Fe found in the substantia nigra of PD patients may perhaps be explained by free Cu binding to iron binding protein-1 (IBP-1), causing it to malfunction and preventing it from detaching itself from the transferrin receptor (TfR) inhibition gene, resulting in expression of TfR even when the cell has plenty of Fe. The gradual accumulation of Fe and Cu would explain the damage inflicted on the substantia nigra by free radicals catalyzed by these two metals and which is exacerbated by the low levels of CuZnSOD, due to the Zn deficiency mentioned above. Moreover, if this hypothesis is correct, then PD could be used to help discover the gene (or genes) responsible for WD and vice versa. Furthermore, idiopathic PD could be prevented by identifying the heterozygote individuals and providing them with Zn supplementation, Cu chelation therapy and phlebotomy to eliminate Fe. PMID- 11425283 TI - The beneficial, antimicrobial effect of probiotics. PMID- 11425284 TI - New strategies in antimicrobial therapy. PMID- 11425285 TI - Can melatonin regulate the expression of prohormone convertase 1 and 2 genes via monomeric and dimeric forms of RZR/ROR nuclear receptor, and can melatonin influence the processes of embryogenesis or carcinogenesis by disturbing the proportion of cAMP and cGMP concentrations? Theoretic model of controlled apoptosis. AB - The presented model of controlled apoptosis has been based on the assumption that correct information exchange between an organism as a whole, and each of its cells is conditioned by mutual proportions of cAMP and cGMP concentrations (CcAMP, CcGMP), according to the formula CcAMP x CcGMP = 'a' (constant). The regulation of balance of these 'second messengers' in a cell and an extracellular space would depend on the mutual proportions of concentrations of Melatonin and monomers of Melanin. These indoloderived compounds could be the activators of the transcription factors i.e. RZR and NFkappa-B, regulating the expression of Prohormone Convertase (PC) gen and Nitric Oxide Synthase (NOS) gen, respectively. Additionally, maternal Melatonin and Nitric Oxide (NO), being able to pass through trophoblast or placenta freely, would play decisive role in the synchronization of embryogenesis and intrauterine development of the fetus. In case of an embryo or a fetus, the result of CcAMP and CcGMP multiplication, different from the proper constant 'a'-value, would mean occurrence of disorders in the structure and functioning of the cellular tensegrity system and, in consequence, disturbances in the intercellular information exchange. It would lead to deviation in cellular metabolism, oriented cell movement, uncontrolled apoptosis, and as a consequence, would lead to the development of fetal defects. In case of a child or an adult, a sudden occurrence and prolongation of such disturbances in CcAMP-CcGMP proportions would induce a process of apoptosis of normal cells and an initiation of a cancerogenesis. On the other hand, the recovery of equilibrium in the information exchange system would initiate apoptosis of neoplastic cells, and simultaneously, proliferation of connective tissue cells. According to the presented hypothesis, a decrease in CcAMP and destabilization of the CcAMP-CcGMP balance in an embryo or a fetus would result from relatively excessive amounts of maternal Melatonin (monomers) in fetal circulation, while a decrease of CcAMP and destabilization of the CcAMP-CcGMP balance in a child or an adult would be a consequence of relatively insufficient amounts of Melatonin (dimers) in an organism. It seems possible, that determination of both CcAMP and CcGMP would enable an early detection of high risk of developmental defects occurrence in an embryo or a fetus and neoplastic processes in a child or an adult. This method might also be considerably useful in monitoring a safe substitutional hormonotherapy. PMID- 11425286 TI - Genomic multi-level nutrient-sensing pathways. AB - Traditionally, the pharmaceutical and natural products industries have looked for products, which affect some biochemical or cellular pathway in either a positive or negative manner. While both industries have enjoyed success with this approach, there is a vast area that as of yet has remained relatively untapped: the genome. The following work explores how both industries can benefit from the burgeoning area of nutrients directed toward the genome. Nutritional supplements capable of altering the integrity, expression, or fidelity of genes are herein referred to collectively as 'genomeceuticals' for their ability to acting on the genome in a manner related to pharmaceuticals acting on biochemical pathways. Central to the paper is the finding that glucosamine can up-regulate the obese (ob) gene by acting on a 'nutrient-sensing' pathway (1). An explanation of how nutrient-sensing pathways might be exploited at the genetic level is presented. Discussion is given to how such genomeceuticals can not just replace substances, which may be missing (e.g., an enzyme diminished by mutation), but actually alter the expression and functionality of gene products in, and resulting from, genomic nutrient-sensing pathways. PMID- 11425287 TI - Multiple factor indices of protection or risk towards disease. AB - In order to combine several factors entailing protection or risk towards disease and to calculate a Protection Multiple Factor Index (PMFI) or, conversely, a Risk Multiple Factor Index (RMFI), we propose the following formulae: (1) PMFI = 2/[(mF)2 + 1] and (2) RMFI = 2/[(imF)2 + 1], where mF is the mean value of the factors considered and imF is the inverse (or reciprocal) of mF. In calculating mF, the value of each 'risk factor' observed in the patient under study (Vp) is expressed by taking the mean normal value (Vmn) as the unit, i.e. by calculating the ratio Vp/Vmn, whereas each 'protection factor' is expressed as the reciprocal of this ratio, i.e. as Vmn/Vp. The 'weight' of the various factors can be changed through multiplication by a number > 1 or < 1. Values of both PMFI and RMFI are always close to 1 in normal subjects, with extreme variations among patients between 0 and 2. The sum of the values of PMFI and RMFI is always equal to 2, so that one index can be deduced from the other. When factors are only two (F1 and F2), the formulae may be simplified as follows: PMFI = 2/[F1 x F2) + 1] and RMFI = 2/[(iF1 x iF2) + 1], where iF = 1/F, with only minimal changes in results. PMID- 11425288 TI - Memory, learning and the brain: an unconventional view. AB - This paper proposes an unconventional theory on the psychophysiological system of learning and memory and of the brain's task. Learning will be considered as the creation of potential memory schemata in the brain. The brain is regarded as a relay station. Psychological experiences and behavioral expressions are considered as based on (not identified with) physiological activities. PMID- 11425289 TI - Androgenic progestins amplify the breast cancer risk associated with hormone replacement therapy by boosting IGF-I activity. AB - Recent epidemiology indicates that unopposed oral estrogen replacement therapy has a surprisingly small impact on breast cancer risk--little if any in overweight women--whereas combined regimens featuring synthetic progestins are attended by a much larger increase in this risk. These findings may reflect the fact that oral estrogen acts on the liver to down-regulate systemic IGF-I activity, whereas concurrent administration of androgens--including the androgenic progestins often used in replacement therapy--abrogates this effect. Increased systemic IGF-I activity has been linked to increased breast cancer risk, and may be largely responsible for the greater incidence of breast cancer in overweight postmenopausal women--who thus should have the most to gain from suppression of IGF-I activity by oral estrogen. Down-regulation of IGF-I may likewise account for the marked reduction in colon cancer risk associated with current estrogen replacement therapy. Fortunately, natural progesterone--now available in micronized oral preparations--does not oppose the hepatic effects of oral estrogen, and moreover may be preferable to androgenic progestins with respect to vascular function. Oral replacement therapy featuring micronized progesterone, if administered throughout postmenopausal life, can be expected to have a highly positive impact on vascular health, bone density, and risks for Alzheimer's disease and colon cancer--benefits which, in most women, may vastly outweigh the associated increase in risk for breast and endometrial cancers. PMID- 11425290 TI - The chlorophyll metabolite phytanic acid is a natural rexinoid--potential for treatment and prevention of diabetes. AB - Synthetic ligands of the retinoid X receptor (RXR) have shown antidiabetic activity in mice, apparently owing to the fact that they stimulate the transcriptional activity of PPAR-gamma/RXR heterodimers, much like thiazolidinedione drugs. The chlorophyll metabolite phytanic acid has been shown to be a natural ligand for RXR, active in concentrations near its physiological levels. It is thus reasonable to suspect that phytanic acid may have utility for treatment and prevention of human type 2 diabetes. Phytanic acid may mimic or complement various effects of conjugated linoleic acids, which have been shown to activate PPAR-gamma/RXR and prevent rodent diabetes. Administration of hydrolyzed chlorophyll may represent the most cost-effective strategy for raising human tissue levels of phytanic acid. PMID- 11425291 TI - Hepatic monitoring of essential amino acid availability may regulate IGF-I activity, thermogenesis, and fatty acid oxidation/synthesis. AB - Diets that are low in certain essential amino acids (EAAs), whether owing to low protein content or poor protein quality, tend to down-regulate systemic IGF-I activity, boost thermogenesis, and suppress hepatic capacity for lipogenesis, while promoting hepatic fatty acid oxidation. It is proposed that for each EAA there is a regulatory protein in hepatocytes whose activity is repressed by adequate levels of its EAA; if one (or more) of these regulatory proteins is active, it serves as a signal of EAA deficiency which then mediates the aforementioned effects on IGF-I activity, thermogenesis, and hepatic fatty acid metabolism. Mechanisms which monitor EAA availability likewise play a role in appetite regulation, thus accounting for the fact that spontaneous calorie consumption tends to be lower on high-protein diets. Diets low in protein quantity or quality may decrease insulin secretion, an effect which should contribute to their impact on IGF-I activity and lipid metabolism. The fact that vegans ingest diets that tend to be relatively low in certain EAAs may play a key role in their characteristic leanness and their decreased risk for diabetes, coronary disease, and cancer. PMID- 11425292 TI - Monoamine oxidase inhibitors for IgA nephropathy. AB - IgA nephropathy (IgAN), characterized by renal mesangial deposits of antibodies (of the IgA subtype), is the most common glomerulonephritis worldwide. The cause of IgAN is not known. IgAN can often lead to end stage renal disease (ESRD), and there is no known treatment proven to prevent ESRD in IgAN. Long term use of steroids or other immunosuppressant drugs carry severe toxicities and other risks. IgAN patients have high serum levels of tumor necrosis factor-alpha (TNF). Increased monoamine levels, via increased cellular cyclic AMP, can decrease TNF elaboration. Monoamine oxidase inhibitors (MAO-Is) have been found effective in case studies for a number of diseases, e.g. rheumatoid arthritis and Crohn's disease, characterized by high TNF levels. Here I suggest that MAO-Is might be of utility in IgAN by decreasing TNF levels. PMID- 11425293 TI - Xenon as neuroprotectant in acute stroke? AB - Stroke is one of the most common diseases worldwide. A current focus of research is to find neuroprotectants that can be given during acute stroke. One specific target is to find antagonists of the N-methyl-D-aspartate (NMDA) receptor in an attempt to limit glutamate associated neurotoxicity. Xenon has recently been found to be an NMDA receptor antagonist. I suggest that xenon be considered as a neuroprotectant in acute stroke, and discuss methods of testing this proposal. PMID- 11425294 TI - Determinants of the revival of dissection of the human body in the Middle Ages. AB - The author reviews the causes of the rebirth of human dissection in the Middle Ages after a hiatus of 1500 years, that is, after the practice was abandoned in the time following Herophilus of Alexandria in the third century BC. It is concluded that three main factors determined the rebirth: the removal of obstacles (i.e., the devaluation of the body), the stimulus to proceed (i.e., the resuming of the slow progress of medicine toward the scientific mode), and the different value attributed to cadavers of different people, that is, the correspondence between life deeds and the importance of the cadaver (which made the corpses of executed criminals available and pedagogical dissection possible). PMID- 11425295 TI - An exploratory approach to the serotonergic hypothesis of depression: bridging the synaptic gap. AB - In this exploratory review, we attempt to integrate pre and post synaptic theories of the biochemical basis of depression--in particular with regard to 5 HT. We will be providing evidence that in major depressive disorder, there is a continuity of dysfunction of neural function, i.e. pre and post synaptic serotonergic symptoms are affected. Furthermore, we will also be providing the implications of this approach for normal treatments for depressive disorder. PMID- 11425296 TI - Panic attack and SIDS: possible therapeutic conclusion. AB - Characteristic features of the panic patients is the increased sensitivity to the CO2. Respiratory pattern abnormalities such as prolonged apnea, diminished ventilatory responsiveness to hypercarbia have been observed in infants at risk for SIDS. Treating the panic attack the SSRI preparations proved to be suitable. Serotonin antagonists effect preventing SIDS is suggested. PMID- 11425297 TI - Autism and Parkinson's disease. AB - The pathogenesis of Parkinson's disease, a neurodegenerative disorder, is multifaceted, having a variety of genetic and environmental factors. There is considerable evidence to support the role of toxins, particularly pesticides and herbicides, in at least some of those affected (presumably, mostly the genetically vulnerable). The pathogenesis of autism is no less complex, but little is known about the potential role of toxins for autism, a neurodevelopmental disorder. The incidence of autism appears to be rising, and early exposure to synthetic chemicals is one suspect for this rise. Impaired detoxification of certain chemicals may be common to autism and Parkinson's disease. Further study of environmental influences for either disorder may lead to important insights regarding causation for both, and perhaps for other neurodegenerative and neurodevelopmental disorders as well. PMID- 11425298 TI - Anergy and acquired immune deficiency syndrome. AB - Human immunodeficiency virus (HIV), the retrovirus associated with acquired immune deficiency syndrome (AIDS), acts as a super-antigen by binding to the variable region of the beta (V beta) chain of T-cell receptor (TCR). It's binding to CD4 molecules and chemokine receptors induces a spectrum of immune abnormalities including 'a state of anergy' in the host. This state is due to a defective function of T-helper cell-1 (Th-1), a reduction in production of lymphokines required for signal transduction, an impaired cytotoxic cell activation and a decrease in antigen presenting function of monocyte-macrophage cell lineage. These immune abnormalities form the basis for severe opportunistic infections and malignancies in the host. Malnutrition, micronutrient abnormalities, concomitant infections and genetic factors, etc., are some of the compounding co-factors that further contribute to 'the state of anergy'. PMID- 11425299 TI - Sudden infant death syndrome (SIDS): T-cell immunodeficiency--Part 1. AB - It is hypothesized that SIDS mimics AIDS and atopic eczema in that defective T lymphocytes and overactive B cells overstimulate pro-inflammatory cytokines in the mucosal immune system. Virally infected cells are unable to convert linoleic acid (LA) into gamma-linolenic acid (GLA) which eventually leads to defective T lymphocyte production. Abnormal lung cytokine synthesis by virus-induced immunodeficient T lymphocytes is associated with the murine AIDS-related complex (ARC). Adenosine triphosphate (ATP) deficient anaerobic cells cannot convert LA to GLA. It is hypothesized that, in SIDS victims, elevated levels of hypoxanthine and immunoglobulins are evidence of chronic hypoxemia and ATP catabolism, and an inability to convert LA to GLA, leading to defective T lymphocytes in the mucosal immune system. PMID- 11425300 TI - Possible involvement of sulfane sulfur in homocysteine-induced atherosclerosis. AB - Homocysteinemia, first identified as a genetic disease in children in the 1960s, is associated with severe widespread atherosclerosis which causes death (in untreated cases) before the age of 10 years. Elevated blood homocysteine is now recognized as a risk factor for heart disease in the general population. The mechanism by which homocysteine induces atherosclerosis is still unknown despite intensive investigation. It is proposed here that the mechanism involves sulfane sulfur formed in the catabolism of homocysteine. This unstable and reactive form of sulfur is formed through the action of several enzymes which are known to use homocysteine, its disulfide (homocystine), or its mixed disulfide with cysteine as substrates. Sulfane sulfur has physiological effects which are consistent with a role in atherogenesis. At very low concentrations, it induces proliferation of many cell types, an effect which is consistent with the fibrosis and hyperplasia, which are prominent features of atherosclerotic lesions. At higher concentrations, it is toxic. Also, it modulates the activity of many enzymes and, through this effect on enzymes of lipid metabolism, it could be responsible for the lipid accumulation seen in atherosclerotic lesions. PMID- 11425301 TI - beta-casein A1, ischaemic heart disease mortality, and other illnesses. AB - The risk factors identified with cardiovascular disease studied in the WHO MONICA project have been shown to have a limited relationship with the coronary heart disease mortality rates between centres, and in mirroring the historical rise and decline in deaths from the disease. Here we show that correlation of the calculated consumption of the milk protein, beta-casein A1 (excluding milk protein in cheese) against ischaemic heart disease (IHD) mortality has a r2 = 0.86. In the states of the former West Germany, where the breed composition of regional cattle herds has remained virtually constant since the 1950s, IHD mortality by state correlates with the estimated consumption of beta-casein A1. Information on other recognized dietary risk factors does not indicate any significant regional difference. Similarly, the populations of Toulouse in France and Belfast in Northern Ireland have almost identical collective 'traditional' risk factors for heart disease, yet the respective mortality rates vary more than threefold. People from Northern Ireland are estimated to consume 3.23 times more beta-casein A1, excluding cheese, than the French. The remarkable agreement between mortality and the consumption of this allele suggests that this factor is worthy of serious consideration as a potential source of cardiovascular disease when taken in conjunction with regional variations in the traditional risk factors. beta-casein A1 consumption also correlates strongly with type 1 diabetes incidence in 0-14-year-olds, suggesting that IHD and diabetes may share at least one causative risk factor. PMID- 11425303 TI - Characterization of an allylic analogue of the 5'-deoxyadenosyl radical: an intermediate in the reaction of lysine 2,3-aminomutase. AB - An allylic analogue of the 5'-deoxyadenosyl radical has been characterized at the active site of lysine 2,3-aminomutase (LAM) by electron paramagnetic resonance (EPR) spectroscopy. The anhydroadenosyl radical, 5'-deoxy-3',4'-anhydroadenosine 5'-yl, is a surrogate of the less stable 5'-deoxyadenosyl radical, which has never been observed but has been postulated to be a radical intermediate in the catalytic cycles of a number of enzymes. An earlier communication [Magnusson, O.Th., Reed, G. H., and Frey, P. A. (1999) J. Am. Chem. Soc. 121, 9764-9765] included the initial spectroscopic identification at 77 K of the radical, which is formed upon replacement of S-adenosylmethionine by S-3',4' anhydroadenosylmethionine as a coenzyme for LAM. The electron paramagnetic resonance spectrum of the radical changes dramatically between 77 and 4.5 K. This unusual temperature dependence is attributed to a spin-spin interaction between the radical and thermally populated, higher spin states of the [4Fe-4S]+2 center, which is diamagnetic at 4.5 K. The EPR spectra of the radical at 4.5 K have been analyzed using isotopic substitutions and simulations. Analysis of the nuclear hyperfine splitting shows that the unpaired spin is distributed equally between C5'- and C3'- as expected for an allylic radical. Hyperfine splitting from the beta-proton at C-2'(H) shows that the dihedral angle to the p(z)-orbital at C-3' is approximately 37 degrees. This conformation is in good agreement with a structural model of the radical. The rate of formation of the allylic radical shows that it is kinetically competent as an intermediate. Measurements of 2H kinetic isotope effects indicate that with lysine as the substrate, the rate limiting steps follow initial reductive cleavage of the coenzyme analogue. PMID- 11425302 TI - Advances in determination of a high-resolution three-dimensional structure of rhodopsin, a model of G-protein-coupled receptors (GPCRs). PMID- 11425304 TI - Detection of protein oxidation in rat-1 fibroblasts by fluorescently labeled tyramine. AB - Oxidative damage to proteins has been postulated as a major cause of various degenerative diseases including the loss of functional capacity during aging. A prominent target for oxidation by reactive oxygen species (ROS) is the tyrosine residue. Here we present a highly sensitive method for the detection of tyrosyl radical formation in cells. The method is based on the fluorescein-labeled tyrosine analogue, tyramine, which upon oxidation may couple to proteins carrying a tyrosyl radical. Coupling of the probe (denoted TyrFluo) to standard proteins could be induced by generating ROS with horseradish peroxidase/hydrogen peroxide, SIN-1 or with peroxides (cumene or hydrogen peroxide) in combination with a transition metal. TyrFluo added to rat-1 fibroblasts remained outside the cell, whereas the acetylated form (acetylTyrFluo) was membrane-permeable and accumulated in the cell. Exposure of the cells to oxidative stress in the presence of either TyrFluo or acetylTyrFluo gave a cellular labeling characteristic for each probe. Western blot analysis confirmed that each probe labeled a specific set of proteins. This new method for the detection of ROS induced oxidation of proteins may mimic the tendency of oxidized proteins to form dityrosine bonds. PMID- 11425305 TI - Polysaccharide recognition by surfactant protein D: novel interactions of a C type lectin with nonterminal glucosyl residues. AB - Surfactant protein D (SP-D), a C-type lectin, is an important pulmonary host defense molecule. Carbohydrate binding is critical to its host defense properties, but the precise polysaccharide structures recognized by the protein are unknown. SP-D binding to Aspergillus fumigatus is strongly inhibited by a soluble beta-(1-->6)-linked but not by a soluble beta-(1-->3)-linked glucosyl homopolysaccharide (pustulan and laminarin, respectively), suggesting that SP-D recognizes only certain polysaccharide configurations, likely through differential binding to nonterminal glucosyl residues. In this study we have computationally docked alpha/beta-D-glucopyranose and alpha/beta-(1-->2)-, alpha/beta-(1-->3)-, alpha/beta-(1-->4)-, and alpha/beta-(1-->6)-linked glucosyl trisaccharides into the SP-D carbohydrate recognition domain. As with the mannose binding proteins, we found significant hydrogen bonding between the protein and the vicinal, equatorial OH groups at the 3 and 4 positions on the sugar ring. Our docking studies predict that alpha/beta-(1-->2)-, alpha-(1-->4)-, and alpha/beta (1-->6)-linked but not alpha/beta-(1-->3)-linked glucosyl trisaccharides can be bound by their internal glucosyl residues and that binding also occurs through interactions of the protein with the 2- and 3-equatorial OH groups on the glucosyl ring. By using various soluble glucosyl homopolysaccharides as inhibitors of SP-D carbohydrate binding, we confirmed the interactions predicted by our modeling studies. Given the sequence and structural similarity between SP D and other C-type lectins, many of the predicted interactions should be applicable to this protein family. PMID- 11425306 TI - Crystal structure of rabbit phosphoglucose isomerase complexed with its substrate D-fructose 6-phosphate. AB - Phosphoglucose isomerase (PGI, EC 5.3.1.9) catalyzes the interconversion of D glucose 6-phosphate (G6P) and D-fructose 6-phosphate (F6P) and plays important roles in glycolysis and gluconeogenesis. Biochemical characterization of the enzyme has led to a proposed multistep catalytic mechanism. First, the enzyme catalyzes ring opening to yield the open chain form of the substrate. Then isomerization proceeds via proton transfer between C2 and C1 of a cis enediol(ate) intermediate to yield the open chain form of the product. Catalysis proceeds in both the G6P to F6P and F6P to G6P directions, so both G6P and F6P are substrates. X-ray crystal structure analysis of rabbit and bacterial PGI has previously identified the location of the enzyme active site, and a recent crystal structure of rabbit PGI identified Glu357 as a candidate functional group for transferring the proton. However, it was not clear which active site amino acid residues catalyze the ring opening step. In this paper, we report the X-ray crystal structure of rabbit PGI complexed with the cyclic form of its substrate, D-fructose 6-phosphate, at 2.1 A resolution. The location of the substrate relative to the side chains of His388 suggest that His388 promotes ring opening by protonating the ring oxygen. Glu216 helps to position His388, and a water molecule that is held in position by Lys518 and Thr214 accepts a proton from the hydroxyl group at C2. Comparison to a structure of rabbit PGI with 5PAA bound indicates that ring opening is followed by loss of the protonated water molecule and conformational changes in the substrate and the protein so that a helix containing amino acids 513-520 moves in toward the substrate to form additional hydrogen bonds with the substrate. PMID- 11425307 TI - Dynamics of nitric oxide in the active site of reduced cytochrome c oxidase aa3. AB - Nitric oxide (NO) is involved in the regulation of respiration by acting as a competitive ligand for molecular oxygen at the binuclear active site of cytochrome c oxidase. The dynamics of NO in and near this site are not well understood. We performed flash photolysis studies of NO from heme a3 in cytochrome c oxidase from Paracoccus denitrificans, using femtosecond transient absorption spectroscopy. The formation of the product state--the unliganded heme a3 ground state--occurs in a similar stepwise manner (period approximately 700 fs) as previously observed for carbon monoxide photolysis from this enzyme and interpreted in terms of ballistic ligand motions in the active site on the subpicosecond time scale [Liebl, U., Lipowski, G., Negrerie, M., Lambry, J.-C., Martin, J.-L., and Vos, M. H. (1999) Nature 401, 181-184]. A fraction (approximately 35% at very low NO concentrations) of the dissociated NO recombines with heme a3 in 200-300 ps. The presence of this recombination phase indicates that a transient bond to the second ligand-binding site, a copper atom (CuB), has a short lifetime or may not be formed. Increasing the NO concentration increases the recombination yield on the hundreds of picoseconds time scale. This effect, unprecedented for heme proteins, implies that, apart from the one NO molecule bound to heme a3, a second NO molecule can be accommodated in the active site, even at relatively low (submicromolar) concentrations. Models for NO accommodation in the active site, based on molecular dynamics energy minimizations are presented. Pathways for NO motion and their relevance for the regulation of respiration are discussed. PMID- 11425308 TI - Vesicle permeabilization by protofibrillar alpha-synuclein: implications for the pathogenesis and treatment of Parkinson's disease. AB - Fibrillar alpha-synuclein is a component of the Lewy body, the characteristic neuronal inclusion of the Parkinson's disease (PD) brain. Both alpha-synuclein mutations linked to autosomal dominant early-onset forms of PD promote the in vitro conversion of the natively unfolded protein into ordered prefibrillar oligomers, suggesting that these protofibrils, rather than the fibril itself, may induce cell death. We report here that protofibrils differ markedly from fibrils with respect to their interactions with synthetic membranes. Protofibrillar alpha synuclein, in contrast to the monomeric and the fibrillar forms, binds synthetic vesicles very tightly via a beta-sheet-rich structure and transiently permeabilizes these vesicles. The destruction of vesicular membranes by protofibrillar alpha-synuclein was directly observed by atomic force microscopy. The possibility that the toxicity of alpha-synuclein fibrillization may derive from an oligomeric intermediate, rather than the fibril, has implications regarding the design of therapeutics for PD. PMID- 11425309 TI - NMR solution structure and backbone dynamics of the CC chemokine eotaxin-3. AB - Eotaxin-3 is one of three related chemokines that specifically activate chemokine receptor CCR3. We report the 3D structure and backbone dynamics of eotaxin-3 determined by NMR spectroscopy. Eotaxin-3 is monomeric under the conditions in this study and consists of an unstructured N-terminus before the first two conserved cysteine residues, an irregularly structured N-loop following the second conserved cysteine, a single turn of 3(10)-helix, a three-stranded antiparallel beta-sheet, an alpha-helix, and an unstructured C-terminal tail. As in other chemokines, the alpha-helix packs against one face of the beta-sheet. The average backbone and heavy atom rmsd values of the 20 structures (residues 9 65) are 0.44 and 1.01 A, respectively. A comparison between the structures of eotaxin-3 and related chemokines suggests that the electrostatic potential in the vicinity of a surface groove and the structure of the beta2-beta3 turn may be important for maintaining receptor specificity. The backbone dynamics of eotaxin 3 were determined from 15N NMR relaxation data using the extended model free dynamics formalism. Large amplitude motions on the picosecond to nanosecond time scale were observed in both termini and in some residues in the N-loop, the beta1 beta2 turn, and the beta3 strand; the location of these residues suggests a possible role for dynamics in receptor binding and activation. In contrast to eotaxin, eotaxin-3 exhibits no substantial mobility on the microsecond to millisecond time scale. PMID- 11425310 TI - The photobleaching sequence of a short-wavelength visual pigment. AB - The photobleaching pathway of a short-wavelength cone opsin purified in delipidated form (lambda(max) = 425 nm) is reported. The batho intermediate of the violet cone opsin generated at 45 K has an absorption maximum at 450 nm. The batho intermediate thermally decays to the lumi intermediate (lambda(max) = 435 nm) at 200 K. The lumi intermediate decays to the meta I (lambda(max) = 420 nm) and meta II (lambda(max) = 388 nm) intermediates at 258 and 263 K, respectively. The meta II intermediate decays to free retinal and opsin at >270 K. At 45, 75, and 140 K, the photochemical excitation of the violet cone opsin at 425 nm generates the batho intermediate at high concentrations under moderate illumination. The batho intermediate spectra, generated via decomposing the photostationary state spectra at 45 and 140 K, are identical and have properties typical of batho intermediates of other visual pigments. Extended illumination of the violet cone opsin at 75 K, however, generates a red-shifted photostationary state (relative to both the dark and the batho intermediates) that has as absorption maximum at approximately 470 nm, and thermally reverts to form the normal batho intermediate when warmed to 140 K. We conclude that this red-shifted photostationary state is a metastable state, characterized by a higher-energy protein conformation that allows relaxation of the all-trans chromophore into a more planar conformation. FTIR spectroscopy of violet cone opsin indicates conclusively that the chromophore is protonated. A similar transformation of the rhodopsin binding site generates a model for the VCOP binding site that predicts roughly 75% of the observed blue shift of the violet cone pigment relative to rhodopsin. MNDO-PSDCI calculations indicate that secondary interactions involving the binding site residues are as important as the first-order chromophore protein interactions in mediating the wavelength maximum. PMID- 11425311 TI - Reciprocity between O-GlcNAc and O-phosphate on the carboxyl terminal domain of RNA polymerase II. AB - The carboxyl terminal domain of RNA polymerase II has multiple essential roles in transcription initiation, promoter clearance, transcript elongation, and the recruitment of the RNA processing machinery. Specific phosphorylation events are associated with the spatial and temporal coordination of these different activities. The CTD is also modified by beta-O-linked GlcNAc on a subset of RNA Pol II molecules. Using synthetic CTD substrates, we show here that O-GlcNAc and phosphate modification of the CTD are mutually exclusive at the level of the enzymes responsible for their addition. In addition, we show that O-GlcNAc transferase and CTD kinase have different CTD repeat requirements for enzymatic activity. The Km values of the two enzymes for CTD substrates are in a similar range, indicating that neither enzyme has a distinct kinetic advantage. Thus, the in vivo regulation of O-GlcNAc and phosphate modification of the CTD may involve the differential association of these two enzymes with the CTD at specific stages during the transcription cycle. Furthermore, direct competition between OGT and CTD kinase in vivo could generate multiple functionally distinct isoforms of RNA Pol II. Taken together, these results suggest that O-GlcNAc may give rise to additional functional states of RNA Pol II and may coordinate with phosphorylation to regulate class II gene transcription. PMID- 11425312 TI - Peptide nucleic acids are potent modulators of endogenous pre-mRNA splicing of the murine interleukin-5 receptor-alpha chain. AB - Antisense oligonucleotides (ASOs) that bind target pre-mRNA with high affinity have been shown to alter splicing patterns and offer promise as therapeutics. Previous studies have shown that ASOs fully modified with 2'-O-methoxyethyl (2'-O MOE) sugar residues redirect constitutive and alternative splicing of the murine interleukin-5 receptor-alpha (IL-5Ralpha) chain pre-mRNA in cells, resulting in inhibition of the membrane-bound isoform and enhanced expression of the soluble isoform. Here, we show that antisense peptide nucleic acids (PNAs) alter splicing of the IL-5Ralpha pre-mRNA in a fashion similar to their 2'-O-MOE-modified counterparts of the same sequence. Moreover, using PNA as the splicing modulator, the length of the antisense oligomer could be shortened from 20 to 15 nucleobase units to obtain a comparable effect. Treatment of cells with antisense PNA resulted in dose-dependent, specific downregulation of IL-5Ralpha membrane isoform mRNA expression and enhanced levels of the soluble IL-5Ralpha isoform transcript, with an EC50 equivalent to that observed in parallel with the corresponding 2'-O-MOE ASO. The pronounced activity of antisense PNAs in modulating IL-5Ralpha mRNA splicing observed in our study identifies these compounds as a promising new class of lower molecular weight splicing modulators. PMID- 11425313 TI - Differential chromatin association and nucleosome binding of the maize HMGA, HMGB, and SSRP1 proteins. AB - In plants, chromosomal high mobility group (HMG) proteins have been identified in the HMGA family, containing A/T-hook DNA binding motifs, and in the HMGB family, containing an HMG-box DNA binding domain, that are considered architectural factors in chromatin. We have characterized the association of the HMGA protein, five different HMGB proteins, and the structure-specific recognition protein 1 (SSRP1) with maize chromatin by extraction experiments using NaCl, ethidium bromide, spermine, and distamycin A. The difference in the release of the proteins from chromatin by these reagents indicates that they are differentially associated with chromatin. This was confirmed by treatment of chromatin with micrococcal nuclease, demonstrating that the HMGA, HMGB2/3, and SSRP1 proteins are enriched in the highly nuclease-sensitive fraction of chromatin, which is likely to be transcriptionally competent. As examined by electrophoretic mobility shift analyses, the HMGA protein and the proteins containing an HMG domain (HMGB proteins and SSRP1) bind specifically to purified maize mononucleosomes that contain a histone octamer and approximately 165 bp of DNA. The mode of interaction with the nucleosomes differs for HMGA and HMGB proteins. In the case of the HMGB1 protein, the full-length protein is required for specific nucleosome binding, as the individual HMG-box DNA binding domain (which is sufficient for DNA interactions) interacts nonspecifically with the nucleosomes. Collectively, these findings indicate that HMGA, the various HMGB proteins, and SSPR1 are differentially associated with plant chromatin and may act as architectural factors in different nucleoprotein structures. PMID- 11425314 TI - The regulatory domain of the myosin head behaves as a rigid lever. AB - The regulatory domain of the myosin head is believed to serve as a lever arm that amplifies force generated in the catalytic domain and transmits this strain to the thick filament. The lever arm itself either can be passive or may have a more active role storing some of the energy created by hydrolysis of ATP. A structural correlate which might distinguish between these two possibilities (a passive or an active role) is the stiffness of the domain in question. To this effect we have examined the motion of the proximal (ELC) and distal (RLC) subdomains of the regulatory domain in reconstituted myosin filaments. Each subdomain was labeled with a spin label at a unique cysteine residue, Cys-136 of ELC or Cys-154 of mutant RLC, and its mobility was determined using saturation transfer electron paramagnetic resonance spectroscopy. The mobility of the two domains was similar; the effective correlation time (tau(eff)) for ELC was 17 micros and that for RLC was 22 micros. Additionally, following a 2-fold change of the global dynamics of the myosin head, effected by decreasing the interactions with the filament surface (or the other myosin head), the coupling of the intradomain dynamics remained unchanged. These data suggest that the regulatory domain of the myosin head acts as a single mechanically rigid body, consistent with the regulatory domain serving as a passive lever. PMID- 11425315 TI - The predicted ATP-binding domains in the hexose transporter GLUT1 critically affect transporter activity. AB - The glucose transporter GLUT1 has three short amino acid sequences (domains I III) with homology to typical ATP-binding domains. GLUT1 is a facilitative transporter, however, and transports its substrates down a concentration gradient without a specific requirement for energy or hydrolysis of ATP. Therefore, we assessed the functional role of the predicted ATP-binding domains in GLUT1 by site-directed mutagenesis and expression in Xenopus oocytes. For each mutant, we determined the level of protein expression and the kinetics of transport under zero-trans influx, zero-trans efflux, and equilibrium exchange conditions. Although all five mutants were expressed at levels similar to that of the wild type GLUT1, each single amino acid change in domains I or III profoundly affected GLUT1 function. The mutants Gly116-->Ala in domain I and Gly332-->Ala in domain III exhibited only 10-20% of the transport activity of the wild-type GLUT1. The mutants Gly111-->Ala in domain I and Leu336-->Ala in domain III showed altered kinetic properties; neither the apparent Km nor the Vmax for 3-methylglucose transport were increased under equilibrium exchange conditions, and they did not show the expected level of countertransport acceleration. The mutant Lys117-->Arg in domain I showed a marked increase in the apparent Km for 3-methylglucose transport under zero-trans efflux and equilibrium exchange conditions while maintaining countertransport acceleration. These results indicate that the predicted ATP-binding domains I and III in GLUT1 are important components of the region in GLUT1 involved in transport of the substrate and that their integrity is critical for maintaining the activity and kinetic properties of the transporter. PMID- 11425316 TI - Structure-function relationships for inhibitors of beta-amyloid toxicity containing the recognition sequence KLVFF. AB - Beta-amyloid (Abeta), the primary protein component of Alzheimer's plaques, is neurotoxic when aggregated into fibrils. We have devised a modular strategy for generating compounds that inhibit Abeta toxicity. These compounds contain a recognition element, designed to bind to Abeta, linked to a disrupting element, designed to interfere with Abeta aggregation. On the basis of this strategy, a hybrid peptide was synthesized with the sequence KLVFF (residues 16-20 of Abeta) as the recognition element and a lysine hexamer as the disrupting element; this compound protects cells in vitro from Abeta toxicity [Pallitto, M. M., et al. (1999) Biochemistry 38, 3570]. To determine if the length of the disrupting element could be reduced, peptides were synthesized that contained the KLVFF recognition element and a sequence of one to six lysines as disrupting elements. All compounds enhanced the rate of aggregation of Abeta, with the magnitude of the effect increasing as the number of lysines in the disrupting element increased. The greatest level of protection against Abeta toxicity was achieved with compounds containing disrupting elements of three or more lysines in sequence. A peptide with an anionic disrupting element, KLVFFEEEE, had activity similar to that of KLVFFKKKK, in both cellular toxicity and biophysical assays, whereas a peptide with a neutral polar disrupting element, KLVFFSSSS, was ineffective. Protective compounds retained activity even at an inhibitor:Abeta molar ratio of 1:100, making these some of the most effective inhibitors of Abeta toxicity reported to date. These results provide critical insight needed to design more potent inhibitors of Abeta toxicity and to elucidate their mechanism of action. PMID- 11425318 TI - Biosynthesis of pteridines. Stopped-flow kinetic analysis of GTP cyclohydrolase I. AB - GTP cyclohydrolase I catalyzes a mechanistically complex ring expansion affording dihydroneopterin triphosphate from GTP. The inherently slow enzyme reaction was studied under single turnover conditions monitored by multiwavelength ultraviolet spectroscopy. The spectroscopic data array was subjected to singular value decomposition and thereby shown to comprise six significant linearly independent optical processes. The data were fitted to a model of six consecutive unimolecular reaction steps where the first was considered to be reversible. The rate-limiting step was shown to occur rather late in the reaction sequence. PMID- 11425317 TI - Tyrosine phosphorylation regulates manganese superoxide dismutase (MnSOD) RNA binding protein activity and MnSOD protein expression. AB - All cells tested contain a cytosolic protein that binds to a defined region in the 3' untranslated region of manganese superoxide dismutase (MnSOD) RNA; both the MnSOD RNA-binding protein (MnSOD-BP) and the cis element are required for efficient translation of MnSOD RNA [Chung, D. J., Wright, A. E., and Clerch, L. B. (1997) Biochemistry 37, 16298-16306]. This study was designed to test the hypothesis that MnSOD-BP activity is regulated by phosphorylation. When cell extracts from whole rat lung or a rat lung fibroblast cell line, RFL-6, were treated in vitro with a protein tyrosine phosphatase, there was a 4-fold increase in MnSOD-BP activity indicating that MnSOD-BP activity was upregulated by tyrosine dephosphorylation. RFL-6 cells treated in cell culture with herbimycin A or genistein, inhibitors of protein tyrosine kinase, had significantly more MnSOD BP activity than cells treated with diluent. In RFL-6 cells treated with herbimycin A, the increase in MnSOD-BP activity was associated with an increase in the level of MnSOD protein without a change in MnSOD mRNA concentration. We propose that the modulation of MnSOD protein expression by the tyrosine phosphorylation state of MnSOD-BP is a potential therapeutic target for increasing MnSOD activity during periods of oxidative stress. PMID- 11425319 TI - Mapping protein interfaces with a fluorogenic cross-linker and mass spectrometry: application to nebulin-calmodulin complexes. AB - Nebulin is a giant multifunctional protein that is thought to serve as both a length-regulating protein ruler and calcium/CaM-mediated regulatory protein on the thin filaments of the skeletal muscle sarcomere. To define molecular interfaces between nebulin and CaM, we thiolated lysines of CaM and ND66, a four module cloned fragment from the C-terminus of nebulin, with 2-iminothiolane and cross-linked the complex with dibromobimane, which alkylates thiol pairs within approximately 6 A of each other to form a fluorescent adduct. Such a two-stage cross-linking generated mainly 1:1 complexes of ND66 and CaM, with a limited extent of intramolecular cross-linking. In-gel chymotryptic digestion of the dibromobimane-cross-linked complexes yielded peptides that were first screened by HPLC with fluorescence detection and then scored for cross-linking with mass spectrometry. Several inter- and intramolecular sites were identified and confirmed further by ESI-MS/MS experiments, defining molecular interfaces and patterns of protein folding. In particular, five intermolecular cross-linking products of sequences within the region of amino acids 83-99 (YKENMGKGTPLPVTPEM) in ND66 and several sequences of CaM indicate that the nebulin-CaM interface is close to, and may overlap with, the nebulin-actin interface. This proximity suggests a potential competition between CaM and actin for this nebulin interface. Intramolecular cross-linking of amino acids 13-16 (KEAF) and 13-18 (KEAFSL) with amino acids 145-148 (MTAK) and 146-148 (TAK) in CaM suggests the interaction of two lobes across the central helix. The cross-linking of amino acids 1-6 (MKTPEM) with amino acids 114-129 (YKENVGKATATPVTPE) and 115-129 (KENVGKATATPVTPE) in ND66 hints at an association of noncontiguous nebulin modules in solution. PMID- 11425320 TI - Factors determining the orientation of axially coordinated imidazoles in heme proteins. AB - Factors determining conformations of imidazole axially coordinated to heme in heme proteins were investigated by analyzing 693 hemes in 432 different crystal structures of heme proteins from the Protein Data Bank (PDB), where at least one histidine is ligated to heme. The results from a search of the PDB for protein structures were interpreted with molecular force field computations. Analysis of data from these crystal structures indicated that there are two main factors that determine the orientations of imidazole ligated to heme. These are the interactions of imidazole with the propionic acid side chains of heme and with the histidine backbone. From the analysis of the crystal structures of heme proteins, it turned out that the hydrogen bonding pattern is often not decisive, though it is probably used by nature to fine-tune the orientation of imidazole axially ligated to heme. We found that in many heme proteins the NdeltaH group of imidazole ligated to heme can assume a number of different hydrogen bonds and that in mutant structures the orientation of the ligated imidazole often does not change significantly, although the mutant altered the hydrogen bonding scheme involving the imidazole. Data from crystal structures of heme proteins show that there are preferred orientations of imidazoles with respect to heme. Generally, the NdeltaH group of imidazole is oriented toward the propionic acid groups of the heme. In some cases, the NdeltaH group of imidazole is close to only one of the propionic acid groups, but it is practically never oriented in the opposite direction. The imidazole also adopts a preferred orientation with respect to its histidine backbone such that the plane of the imidazole ring is practically never parallel to the Calpha-Cbeta bond of its histidine backbone. For a given conformation of histidine backbone with respect to heme, as well as imidazole with respect to histidine backbone, the orientation of the imidazole with respect to heme is uniquely determined, since the three orientations depend on each other. Hence, the interaction of the imidazole with the backbone also influences the orientation of the imidazole with respect to the heme. Force field computations are in agreement with experimental data. With this method, we showed that there is an energy minimum when the NdeltaH group of the imidazole is oriented toward the propionic acid groups and that there are minima of energy for orientations where the imidazole ring is orthogonal to the plane defined by the Calpha-Cbeta and Cbeta-Cgamma bonds of the histidine. The computations also demonstrated that these interactions are mainly of electrostatic origin. By taking into account these two major factors, we were able to understand the orientations of axially coordinated imidazoles for all groups of heme proteins, except for the group of cytochrome c peroxidase. In this group, the orientation of the imidazole is determined by a strong hydrogen bond of the NdeltaH group with Asp235. PMID- 11425321 TI - Chromophore structure in lumirhodopsin and metarhodopsin I by time-resolved resonance Raman microchip spectroscopy. AB - Time-resolved resonance Raman microchip flow experiments have been performed on the lumirhodopsin (Lumi) and metarhodopsin I (Meta I) photointermediates of rhodopsin at room temperature to elucidate the structure of the chromophore in each species as well as changes in protein-chromophore interactions. Transient Raman spectra of Lumi and Meta I with delay times of 16 micros and 1 ms, respectively, are obtained by using a microprobe system to focus displaced pump and probe laser beams in a microfabricated flow channel and to detect the scattering. The fingerprint modes of both species are very similar and characteristic of an all-trans chromophore. Lumi exhibits a relatively normal hydrogen-out-of-plane (HOOP) doublet at 951/959 cm(-1), while Meta I has a single HOOP band at 957 cm(-1). These results suggest that the transitions from bathorhodopsin to Lumi and Meta I involve a relaxation of the chromophore to a more planar all-trans conformation and the elimination of the structural perturbation that uncouples the 11H and 12H wags in bathorhodopsin. Surprisingly, the protonated Schiff base C=N stretching mode in Lumi (1638 cm(-1)) is unusually low compared to those in rhodopsin and bathorhodopsin, and the C=ND stretching mode shifts down by only 7 cm(-1) in D2O buffer. This indicates that the Schiff base hydrogen bonding is dramatically weakened in the bathorhodopsin to Lumi transition. However, the C=N stretching mode in Meta I is found at 1654 cm(-1) and exhibits a normal deuteration-induced downshift of 24 cm(-1), identical to that of the all-trans protonated Schiff base. The structural relaxation of the chromophore-protein complex in the bathorhodopsin to Lumi transition thus appears to drive the Schiff base group out of its hydrogen-bonded environment near Glu113, and the hydrogen bonding recovers to a normal solvated PSB value but presumably a different hydrogen bond acceptor with the formation of Meta I. PMID- 11425322 TI - Quantifying the ion selectivity of the Ca2+ site in photosystem II: evidence for direct involvement of Ca2+ in O2 formation. AB - Calcium is an essential cofactor in the oxygen-evolving complex (OEC) of photosystem II (PSII). The removal of Ca2+ or its substitution by any metal ion except Sr2+ inhibits oxygen evolution. We used steady-state enzyme kinetics to measure the rate of O2 evolution in PSII samples treated with an extensive series of mono-, di-, and trivalent metal ions in order to determine the basis for the affinity of metal ions for the Ca2+-binding site. Our results show that the Ca2+ binding site in PSII behaves very similarly to the Ca2+-binding sites in other proteins, and we discuss the implications this has for the structure of the site in PSII. Activity measurements as a function of time show that the binding site achieves equilibrium in 4 h for all of the PSII samples investigated. The binding affinities of the metal ions are modulated by the 17 and 23 kDa extrinsic polypeptides; their removal decreases the free energy of binding of the metal ions by 2.5 kcal/mol, but does not significantly change the time required to reach equilibrium. Monovalent ions are effectively excluded from the Ca2+-binding site, exhibiting no inhibition of O2 evolution. Di- and trivalent metal ions with ionic radii similar to that of Ca2+ (0.99 A) bind competitively with Ca2+ and have the highest binding affinity, while smaller metal ions bind more weakly and much larger ones do not bind competitively. This is consistent with a size selective Ca2+-binding site that has a rigid array of coordinating ligands. Despite the large number of metal ions that competitively replace Ca2+ in the OEC, only Sr2+ is capable of partially restoring activity. Comparing the physical characteristics of the metal ions studied, we identify the pK(a) of the aqua ion as the factor that determines the functional competence of the metal ion. This suggests that Ca2+ is directly involved in the chemistry of water oxidation and is not only a structural cofactor in the OEC. We propose that the role of Ca2+ is to act as a Lewis acid, binding a substrate water molecule and tuning its reactivity. PMID- 11425324 TI - Neural signals that regulate GnRH neurones directly during the oestrous cycle. AB - GnRH, produced by a loose network of neurones in the basal forebrain, is the primary brain signal responsible for the release of LH and FSH from the anterior pituitary gland. The ovarian steroid hormone oestradiol feeds back at both the central nervous system and the anterior pituitary to regulate the patterns of release of GnRH and the gonadotrophins. Although recent evidence indicates that oestradiol may act directly on some GnRH neurones through classical genomic mechanisms, data from published studies have demonstrated that neurotransmission of afferent neuronal systems that are receptive to oestradiol is necessary to drive reproductive cyclicity. Many classical neurotransmitters and neuropeptides alter GnRH neuronal activity, through direct and sometimes indirect actions. This review focuses on the neurotransmitters that regulate GnRH neurones by binding to and activating specific membrane receptors that are expressed in GnRH neurones. These include the catecholamines, gamma-aminobutyric acid, glutamate, neuropeptide Y, neurotensin, beta-endorphin and vasoactive intestinal polypeptide. On the basis of recent molecular and neuroanatomical evidence, it is proposed that oestradiol influences the activity of these neurotransmitter and neuropeptide systems within the GnRH network to drive reproductive cyclicity. PMID- 11425325 TI - Comparative aspects of conceptus growth: a historical perspective. AB - This review provides a historical background for a series of articles arising from a symposium on Early Regulation of Mammalian Development. It begins by tracing connections between the work of J. Cossar Ewart and the first cloning of mammals by somatic cell nuclear transfer. Historical scientific landmarks over the last century that have affected our attitudes to 'fetal autonomy' and conceptus development are reviewed briefly, as are salient studies that have established the importance of nutrition to pre- and postnatal life. Finally, attention is drawn to some differences in conceptus development among various mammals, and the argument made that understanding these differences might be of benefit to the development of reproductive technology. PMID- 11425326 TI - Ribosomal RNA gene expression and chromosome aberrations in bovine oocytes and preimplantation embryos. AB - This review focuses on the key features of development of the bovine oocyte and embryo, with comparisons of the developmental characteristics of embryos produced in vivo and in vitro. The oocyte is transcriptionally quiescent in the primordial and primary follicle. In the secondary follicle transcription is initiated in the oocyte and a ribosome-synthesizing nucleolus is established in this cell. Transcription and nucleolar activity are enhanced in the tertiary follicle during oocyte growth. When the oocyte reaches approximately 110 microm in diameter, corresponding to a follicle of about 3 mm in diameter, transcription ceases and the nucleolus is inactivated, forming a dense spherical remnant. During the final phase of follicular dominance this remnant becomes vacuolated and, in conjunction with resumption of meiosis, it disperses. The rRNA genes are apparently re activated during the four-cell stage, that is, the third cell cycle after fertilization, but a nucleolus is not formed. During the subsequent cell cycle, that is, during the eight-cell stage, ribosome-synthesizing nucleoli are again established. Bovine embryos produced in vitro apparently display the same pattern of nucleolus development as that in embryos developed in vivo. Examination of the ploidy of embryonic cells using fluorescence in situ hybridization has revealed that the production of bovine embryos in vitro is associated with increased chromosome aberrations in the embryos. Blastocysts produced in vitro display a significantly higher rate of mixoploidy, that is, when the embryo consists of both normal diploid and abnormal polyploid cells, than that in embryos developed in vivo. The rate of mixoploidy among embryos produced in vitro increases with increasing developmental stage. Moreover, after fertilization in vitro, initially there is a high rate of 'true' polyploidy, that is, when all cells of the embryos are polyploid. However, the polyploid embryos are eliminated before they cleave beyond the eight-cell stage, the stage at which major activation of the embryonic genome occurs in cattle. PMID- 11425327 TI - Insulin-like growth factor axis during embryonic development. AB - The insulin-like growth factor (IGF) axis has been studied extensively in the developing vertebrate embryo. Knockout experiments have demonstrated that both IGF-I and -II are required for normal development in the mouse embryo, and mRNA and protein expression patterns for both growth factors, together with those for the type I IGF receptor and the six IGF-binding proteins, have been analysed in embryos from different species. Although the unique temporal and spatial expression patterns of these genes indicates important roles for the IGF axis during organ and whole animal development, the variation and complexity of expression makes these roles difficult to unravel. However, one possible mechanism unifying the IGF system in development is programmed cell death (apoptosis), which has been shown to be important in sculpting embryonic tissues, and, in particular, the developing limb bud. In addition, the very early onset of expression of various IGF family members in chicken embryos further emphasizes the fundamental importance of this system in development. This article reviews the work that has been carried out in this area in the context of current understanding of the IGF system. PMID- 11425328 TI - Regulation of gene transcription in the epididymis. AB - The epididymis exhibits region-specific as well as cell-specific patterns of gene expression within the epithelium. The spatial and temporal patterns of gene expression originate during development and are critical to the formation and maintenance of a fully functional epididymis. Despite the number of mechanisms reported to contribute to the regulation of eukaryotic gene expression, little is known about the specific mechanisms involved in the control of epididymal gene expression. This review will outline some of the cis-DNA elements and associated transcription factors that have been identified in the epididymis, in addition to discussing the potential role of co-regulator molecules and changes in chromatin structure as critical control points of gene expression. Although gene expression can be controlled at several points, discussion will focus on gene regulation at the transcriptional level. The role of post-transcriptional control, with particular attention to mRNA stability, will also be discussed. PMID- 11425329 TI - Effect of cytokinesis inhibitors, DMSO and the timing of oocyte activation on mouse cloning using cumulus cell nuclei. AB - Cloning methods are now well described and in almost routine use. However, the frequencies of production of live offspring from activated oocytes remain at < 3% and little is known about the factors that affect these frequencies. The effects of cytokinesis inhibitors, dimethylsulphoxide (DMSO) and the cell cycle of recipient cytoplasm on the cloning of mice were examined. Reconstructed oocytes, which were activated immediately after nucleus injection and cultured without cytochalasin B, developed into blastocysts at a frequency of 30--54% and into live cloned offspring at a frequency of 2--3%. Activated zygotes did not support development to full term after nuclear transfer. Reconstructed oocytes were activated 1--3 h after nuclear transfer and were exposed separately to three inhibitors of cytokinesis (cytochalasin B, cytochalasin D or nocodazole) to examine the toxicity of these inhibitors on cloning. All of the oocytes exposed to nocodazole-containing media formed many small pseudo-pronuclei, whereas with cytochalasin-containing media most of the activated oocytes formed only two pseudo-pronuclei. Despite such differences, 42--61% of reconstructed embryos developed to the morula-blastocyst stage and 1--3% developed to full term in all groups. Addition of 1% (v/v) DMSO to the activation medium significantly improved the frequency of development to the blastocyst stage and full term; however, this improvement did not lead to a higher success rate in the generation of live cloned offspring. These results show that activated mouse oocytes/zygotes are not effective cytoplasmic recipients with the methods described and that the lack of success of cloning is not due to inhibition of cytokinesis. PMID- 11425330 TI - A novel murine tryptase involved in blastocyst hatching and outgrowth. AB - Before implantation the blastocyst is maintained within a proteinaceous coat, the zona pellucida, which prevents polyspermy and ectopic pregnancy. An extracellular trypsin-like activity, which is necessary for hatching from the zona pellucida in vitro, is localized to the abembryonic pole of the blastocyst. Upon hatching, the extracellular matrix-degrading proteinases urokinase plasminogen activator (uPA) and matrix metalloproteinase 9 (MMP-9) are thought to promote blastocyst invasion. However, gene disruption experiments have demonstrated that uPA and MMP 9 are dispensable and, thus, that other key enzymes are involved in implantation. In this study, a novel implantation serine proteinase (ISP1) gene, which is distantly related to haematopoietic tryptases and represents a novel branch of the S1 proteinase family, was cloned. ISP1 is expressed throughout morulae and blastocysts during hatching and outgrowth. Abrogation of ISP1 mRNA accumulation using antisense oligodeoxynucleotides disrupts blastocyst hatching and outgrowth in vitro. The results of this study indicate that the ISP1 gene probably encodes the long sought after 'hatching enzyme' that is localized to the abembryonic pole during hatching in vitro. ISP1 is the earliest embryo-specific proteinase to be expressed in implantation and may play a critical role in connecting embryo hatching to the establishment of implantation competence at the abembryonic pole of the blastocyst. PMID- 11425331 TI - Immunolocalization of progesterone receptors in the canine ovary and their relation to sex steroid hormone concentrations. AB - The aim of the present study was to describe the normal cellular distribution of progesterone receptors in the canine ovary at different stages of the oestrous cycle. Samples of both ovaries were obtained from 75 healthy adult bitches of various breeds and ages, including five pregnant bitches and three bitches that had just delivered. The presence of progesterone receptors was visualized by immunohistochemistry on paraffin wax sections using a monoclonal antibody. Nuclear staining for progesterone receptors was observed in the surface epithelium, cortical tubules, rete ovarii, follicle cells, thecal cells, luteal cells, granulosa cell cords and ovarian stroma. The staining intensity for progesterone receptors in the follicle cells increased with the stage of follicle development, indicating an intrafollicular role of progesterone in the mechanism of ovulation and luteinization. The stronger staining intensities for progesterone receptors in thecal cells compared with follicle cells may be explained by the fact that thecal cells mediate some effects of steroid hormones on the follicle cells in secondary and tertiary follicles. Little correlation was found between the expression of progesterone receptors in follicle cells and oestradiol, progesterone or testosterone concentrations. This finding indicates a different regulating mechanism for progesterone receptors in canine ovarian follicles compared with other tissues of the genital tract. During pregnancy all groups of ovarian cells had lower staining intensity scores than during the oestrous cycle, although the sex steroid hormone concentrations in pregnant bitches were similar to those in non-pregnant bitches during the luteal phase of the oestrous cycle. The lower expression of progesterone receptors during pregnancy may be due to higher tissue concentrations of progesterone that are not reflected in the serum because of haemodilution and increased metabolism and clearance during pregnancy. PMID- 11425332 TI - Role of vascular endothelial growth factor in endometrial vascular events before implantation in rats. AB - Two endometrial vascular events, increased permeability and endothelial cell proliferation, characterize the early stages of embryo implantation in rats. Vascular endothelial growth factor (VEGF), a heparin-binding homodimeric glycoprotein, is expressed in the uterus at the time of implantation. The aim of this study was to investigate the effects of systemic administration of blocking antibody to VEGF on endometrial vascular permeability and endothelial cell proliferation around the time of embryo implantation in rats. Rats were injected i.p. with 1.0, 0.8 or 0.6 ml of anti-VEGF antibody or control serum on day 3 of pregnancy and Evans blue dye was administered i.v. on day 5 of pregnancy. The number of implantation sites (blue bands along the uterus) was counted and endothelial cell proliferation was identified using double staining immunohistochemistry. The number of blue bands was significantly lower (P < 0.05) after injection of 1.0 ml blocking antibody (1.8 +/- 1.56) compared with that in control rats (11.6 +/- 1.97). The number of blue bands was significantly different among rats injected with 1.0 (1.8 +/- 1.56), 0.8 (6.0 +/- 3.67) or 0.6 (10.7 +/- 0.33) ml anti-VEGF antibody, indicating a concentration effect of anti VEGF antibody. No significant differences in endothelial cell proliferation at embryo implantation sites were observed after injection with anti-VEGF antibody or control serum. At intersites (uterine zones between the blue bands), injection of 0.6 ml anti-VEGF antibody caused a significant reduction (P < 0.05) in the percentage of vessels with proliferating endothelial cells (7.25 +/- 2.39%), with a non-significant reduction using 1.0 or 0.8 ml anti-VEGF antibody. No significant differences in microvascular density at either embryo implantation sites or intersites were observed between rats injected with anti-VEGF antibody or control serum. In conclusion, VEGF is the main factor responsible for increased endometrial vascular permeability at implantation. It may regulate endometrial endothelial cell proliferation before implantation and at intersites, thus indicating that factors or mechanisms other than VEGF may regulate endothelial cell proliferation at the site of implantation. PMID- 11425333 TI - Role of prostaglandins in the suppression of apoptosis in hen granulosa cells by transforming growth factor alpha. AB - Although transforming growth factor alpha (TGF-alpha) is known to be an important survival factor for granulosa cells, the cellular and molecular mechanisms involved are uncertain. The purpose of the present study was to investigate the possible involvement of prostaglandins in the anti-apoptotic action of TGF-alpha. Hen granulosa cells from healthy prehierarchical follicles (2-6 mm) cultured in serum-free medium underwent spontaneous apoptosis as demonstrated by DNA fragmentation and nuclear chromatin condensation. TGF-alpha (20 ng ml(-1)) stimulated maximum synthesis of prostaglandins (PGE and PGF) in granulosa cells and completely inhibited serum deprivation-induced apoptosis. The addition of an inhibitor of cyclooxygenase (COX; N-(2-cyclohexyloxy-4 nitrophenyl)methanesulfonamide (NS398) or ibuprofen) or phospholipase A(2) (PLA(2); aristolochic acid, 2-p-amylcinnamoyl amino-4-chlorobenzoic acid (ONO-RS 82) or arachidonyl triflouro methyl ketone (TFMK)), to the culture medium markedly suppressed the TGF-alpha-induced prostaglandin synthesis and significantly increased granulosa cell apoptosis. The apoptotic effect of NS398 and aristolochic acid was completely inhibited by exogenous prostaglandins (PGF(2 alpha), PGE(1), PGE(2)) and arachidonic acid, respectively. However, exogenous prostaglandins failed to inhibit the PLA(2) inhibitor-induced apoptotic DNA fragmentation, implying that in addition to prostaglandins, arachidonic acid or leukotrienes may be important in transducing the anti-apoptotic action of TGF alpha. In the absence of exogenous TGF-alpha, prostaglandins had no significant influence on granulosa cell apoptosis induced by serum withdrawal. These findings indicate that prostaglandin synthesis is a necessary, but not sufficient, event in the suppression of granulosa cell apoptosis by TGF-alpha. Whether arachidonic acid or leukotrienes are important in the anti-apoptotic action of TGF-alpha in hen granulosa cells remains to be determined. PMID- 11425334 TI - Effect of LH on circulating oestradiol and follicular fluid factor concentrations during follicle deviation in cattle. AB - Progesterone was used to reduce LH concentrations starting at the time when the largest follicle was > or = 5.7 mm in diameter or well before the expected start of follicle deviation (largest follicle > or = 8.5 mm in diameter). Plasma concentrations of LH, FSH and oestradiol were determined at 4 h intervals in control and progesterone-treated heifers (n = 8 per group). Concentrations of LH were lower (P < 0.05) in the progesterone-treated group, reflecting an absence of the transient increase in LH concentrations that encompasses follicle deviation. An increase in oestradiol and a continued decrease in FSH occurred at the start of follicle deviation in the control cows but not in the treated heifers. In a second experiment, follicular fluid of the two largest follicles of control and progesterone-treated heifers was sampled at the expected start of deviation (n = 8--10 per group). The concentrations of oestradiol, but not androstenedione and free insulin-like growth factor I (IGF-I), in follicular fluid were higher (P < 0.001) in the largest follicle than in the second largest follicle. Progesterone treatment reduced (P < or = 0.02) the concentrations of all three factors in follicular fluid and increased (P < 0.05) the concentrations of insulin-like growth factor binding protein 2 (IGFBP-2). These results confirm that oestradiol contributes to the continued decrease in FSH concentrations after the start of follicle deviation. Prevention of the transient LH increase, the oestradiol increase and the continued FSH decrease did not significantly alter the mean time or follicle diameters characteristic of expected follicle deviation. However, in some treated individuals (three of eight), the observed follicle deviation was delayed. In addition, these results indicate that the secretion of oestradiol into the circulation and the increase in oestradiol and IGF-I and decrease in IGFBP-2 concentrations in the follicular fluid at the start of deviation are functions of the transient increase in LH concentrations that encompasses follicle deviation. PMID- 11425335 TI - Gonadotrophin-induced follicle development in red deer hinds during the breeding and non-breeding seasons. AB - The effect of exogenous equine chorionic gonadotrophin (eCG) and endogenous (GnRH treatment) gonadotrophins in promoting antral follicle development in red deer hinds was assessed during the breeding and non-breeding seasons. After progesterone pretreatment, hinds received no further treatment, a single injection of 300 iu equine chorionic gonadotrophin or infusion of GnRH (1.0 microg h(-1)) for up to 4 days. Ovaries were recovered (n = 5 per group) at the time of, or 36 h after, progesterone removal. All follicles > or = 2.0 mm in diameter were dissected out, their health status assessed and follicular fluid oestradiol content measured. Granulosa cells were counted and their capacity for oestradiol and cAMP production assessed in vitro. Oestrus, the preovulatory LH surge and ovulation rates were monitored in additional groups of hinds that had been treated identically (n = 5 per group). During the breeding season, all monitored animals ovulated, but five of 15 (one eCG, three GnRH, one progesterone alone) did not exhibit oestrus, and three (all eCG) had luteinized follicles. During seasonal anoestrus, four hinds (all eCG-treated) displayed oestrus, but only two ovulated. Two non-ovulating hinds (one eCG, one progesterone alone) had luteinized follicles. The total number of follicles, or of large (> or = 4 mm) follicles, did not differ significantly between seasons. There were proportionately more healthy follicles during seasonal anoestrus (P < 0.05). Treatment with GnRH, and to a lesser extent eCG, increased the number of oestrogenic follicles and their follicular fluid oestradiol content. In summary, exogenous and endogenous gonadotrophins affected antral follicle development similarly in both the breeding and non-breeding seasons, although the response was variable among animals and often associated with failure of oestrous expression, ovulation and with follicle luteinization. PMID- 11425336 TI - Differential expression of extracellular matrix components in the bovine oviduct during the oestrous cycle. AB - Components of the extracellular matrix take part in tissue rebuilding as well as activating surface-bound growth factors. In the present study, expression and selected activities of urokinase-type plasminogen activator (uPA), matrix metalloproteinases (MMPs), their inhibitors (plasminogen activator inhibitor 1 (PAI-1) and tissue inhibitors of metalloproteinases (TIMPs)) were examined in bovine oviducts by RT--PCR, ribonuclease protection assay and activity assays. A high content of mRNA encoding for uPA was detected before ovulation with a three fold decrease after ovulation. In contrast, PAI-1 expression appeared to be stable during the oestrous cycle. Oviductal flushings produced caseinolytic zones in zymograms containing plasminogen at approximately 50 kDa and 28 kDa. An activity assay for uPA showed highest net activity during the early to mid-luteal phase. Increased TIMP-1 and MMP-2 mRNA concentrations were found around the time of ovulation compared with the luteal phase. In contrast, MMP-1 mRNA transcripts were enriched during the early to mid-luteal phase. Gelatin zymograms detected a 70--72 kDa protease activity showing an oestrous cycle-dependent activity with highest activity before ovulation. Reverse zymography detecting TIMPs revealed proteins between 21 kDa and 24 kDa. Only for the smallest (21 kDa) protein were amounts increased around the time of ovulation compared with the luteal phase. The observation that several extracellular matrix components were regulated distinctly in bovine oviducts indicates that local interactions between these components, growth factors, gametes and the embryo are possible and may influence fertilization and early embryonic development. PMID- 11425337 TI - Lipid and fatty acid analysis of fresh and frozen-thawed immature and in vitro matured bovine oocytes. AB - The lipid content and fatty acid composition of fresh immature and in vitro matured bovine oocytes cultured in media with or without serum, and also those of frozen-thawed immature oocytes were analysed. All oocytes were ranked (A or B) on the basis of their cytoplasmic quality. Fatty acid composition (mol %; w/w) in the total lipid fraction was analysed by gas chromatography. Triglyceride, total cholesterol, phospholipid (phosphocholine-containing phospholipid) and non esterified fatty acid contents of immature and in vitro matured oocytes were determined using lipid analysis kits. Phosphocholine-containing phospholipid and non-esterified fatty acid contents were determined in frozen-thawed immature bovine oocytes. Palmitic acid was the most abundant fatty acid in immature oocytes (A: 35%, B: 36%), and in in vitro matured oocytes cultured in the medium containing serum (A: 36%, B: 35%) or polyvinyl alcohol (A: 33%, B: 36%). Oleic acid was the second most abundant fatty acid in all A ranked oocytes, whereas stearic acid was the second most abundant fatty acid in all B ranked oocytes. There were significant differences (P < 0.05) in linoleic and arachidonic acid fractions between A and B ranked immature oocytes. In vitro matured oocytes had significantly (P < 0.05) lower proportions of linoleic and arachidonic acids, and significantly (P < 0.01) lower contents of triglyceride and total cholesterol compared with those of immature oocytes. The fatty acid composition of in vitro matured oocytes cultured in medium containing fetal calf serum or polyvinyl alcohol was similar, but significant differences (P < 0.01) in triglyceride and the total cholesterol content were observed. There was a significant decrease (P < 0.05) in the arachidonic acid proportion in frozen-thawed immature oocytes compared with that in fresh immature oocytes. In addition, significant (P < 0.05) decreases in both phospholipid (15.8--10.6 pmol) and non-esterified fatty acid (11.0--4.1 pmol) were found in frozen--thawed immature oocytes. The results indicate that lipids are available for use as an energy source for maturation and that serum lipids are incorporated into the oocyte cytoplasm during in vitro maturation. The changes in the lipid content (mainly phospholipid) and fatty acid composition were also observed in frozen--thawed immature oocytes. The study indicates that the alteration of fatty acid composition in bovine oocytes might improve maturation and cryopreservation. PMID- 11425338 TI - Characterization of angiotensin-converting enzyme in canine testis. AB - The aim of this study was to characterize angiotensin-converting enzyme (ACE) in canine testis. Detergent-extracted canine testes were sonicated in the presence of protease inhibitors and purified on an affinity column with the ACE inhibitor, lisinopril, as an affinity ligand for ACE. The fractions recovered were assessed for ACE enzyme activity via an enzyme kinetic microplate assay (at 330 nm) based on the hydrolysis of Fa-Phe-Gly-Gly (FAPGG) at pH 7.5 during an 8 min incubation. The specific activity of ACE in the starting testicular extracts was 3.53 +/- 0.99 mU mg(-1) protein with a 1588 times enrichment in ACE activity after lisinopril affinity chromatography (4239 +/- 2600 mU mg(-1) protein). The recovery efficiency of ACE after lisinopril affinity chromatography was 71.2%. The ACE activity in the detergent extracts and the purified fractions was inhibited significantly by 10 micromol captopril l(-1), a specific ACE inhibitor, and was restored to 88% of normal activity by the addition of the thiol alkylating agent N-ethylmaleimide (0.5 mmol l(-1)) in the detergent extracts and the purified fractions incubated with captopril. The treatment of testicular extracts with 10 mmol EDTA l(-1) reduced the ACE activity significantly (5.40 +/- 1.26 versus 0.58 +/- 0.23 mU mg(-1)). The ACE activity was restored fully in the presence of zinc (5.28 +/- 0.70 mU mg(-1)). The anti-ACE antibody (raised against a 70 kDa protein from the periacrosomal plasma membrane of equine spermatozoa) recognized a 65-70 kDa protein in the detergent-extracted testes as well as in the affinity-purified fractions. This antibody also recognized a protein of similar molecular mass in ejaculated spermatozoa. ACE was localized in the periacrosomal area of the ejaculated spermatozoa and in spermatids in the seminiferous tubules. The results of this study demonstrate that ACE is present in canine testis and retains its enzyme activity after purification with lisinopril affinity chromatography. Activity of canine ACE is inhibited by captopril and EDTA and is restored in the presence of N-ethylmaleimide and zinc. PMID- 11425339 TI - Control of red fox (Vulpes vulpes) fertility with cabergoline: dose response and timing of intervention. AB - Cabergoline, a potent dopamine agonist and inhibitor of prolactin secretion, was investigated as a potential fertility control agent in the red fox ( Vulpes vulpes). Sixty silver fox vixens were selected randomly and artificially inseminated. Cabergoline was fed to groups of 12 vixens in a minced beef ration either as a single dose of 25, 50 or 100 microg kg(-1), or a dose of 50 microg kg(-1) that was repeated 2 days later (2 x 50 microg kg(-1)). Four foxes from each group of 12 were given cabergoline at day 28, day 35 or day 48 after artificial insemination, and a control group of four foxes was used as a comparison for each dose day. In a separate trial, two groups of five foxes were selected randomly from the farm population and fed 100 microg kg(-1) of either cabergoline or a placebo each day from day 42 to day 46 of pregnancy. Foxes that received single doses of cabergoline of 100 microg kg(-1) or 2 x 50 microg kg(-1) aborted at day 28, but the same doses did not result in abortions when administered on days 35 and 48. Although lactation was not terminated in groups that received a single or double dose of cabergoline, increased post-natal cub mortality was associated with cabergoline administration. Growth of cubs between 4 and 8 weeks of age was not inhibited in vixens that received cabergoline. Doses of 100 microg cabergoline kg(-1) administered each day from day 42 to day 46 resulted in abortions and terminated lactation. The capacity of single doses of cabergoline to cause abortions in the red fox during mid- rather than late pregnancy is contrary to reported observations for the domestic dog. This finding indicates that luteotrophic support of the corpus luteum by prolactin may be more important at mid-pregnancy in the red fox. The results of this study support previous field observations that cabergoline delivered in bait affects the reproductive success of vixens and may be a practical adjunct to the lethal control of wild red foxes in Australia. PMID- 11425340 TI - Translocation of active mitochondria during pig oocyte maturation, fertilization and early embryo development in vitro. AB - The distribution of active mitochondria during pig oocyte maturation, fertilization and early embryo development in vitro was revealed by using MitoTracker Green staining and confocal laser scanning microscopy. The regulation of mitochondrial translocation by microfilaments and microtubules was also studied. In oocytes collected from small follicles, strong staining of active mitochondria was observed in the cell cortex. Accumulation of active mitochondria in the peripheral cytoplasm and around the germinal vesicles was characteristic of fully grown oocytes collected from large follicles. Mitochondria accumulated in the perinuclear area during meiotic progression from germinal vesicle breakdown (GVBD) to anaphase I. Larger mitochondrial foci were formed and moved to the inner cytoplasm in mature oocytes. Compared with the oocytes matured in vivo, in which large mitochondrial foci were distributed throughout the cytoplasm, mitochondria were not observed in the central cytoplasm in most of the oocytes matured in vitro. Strong staining of mitochondria was observed in the first polar bodies in metaphase II oocytes. In fertilized eggs, active mitochondria aggregated in the pronuclear region. Perinuclear clustering and a cortical ring were the most marked features of early cleavage. Active mitochondria were distributed in both inner cell mass cells and trophectoderm cells of the blastocysts. Disassembly of microtubules with nocodazole inhibited both mitochondrial aggregations to the germinal vesicle area and their inward movement to the inner cytoplasm during oocyte maturation, as well as the translocation of mitochondria to the peri-pronuclear region during fertilization, whereas disruption of microfilaments by cytochalasin B had no effects. These data indicate that: (i) oocyte maturation, fertilization and early embryo development in pigs are associated with changes in active mitochondrial distribution; (ii) mitochondrial translocation is mediated by microtubules, but not by microfilaments; and (iii) in vitro maturation conditions may cause incomplete movement of mitochondria to the inner cytoplasm and thus affect cytoplasmic maturation. PMID- 11425341 TI - Glycoconjugates on the surface of epididymal spermatozoa in a marsupial, the brushtail possum, Trichosurus vulpecula. AB - Variation in localization and distribution of saccharides on the sperm surface of a marsupial, the brushtail possum, Trichosurus vulpecula, was compared between spermatozoa from the caput and cauda epididymides. Spermatozoa were subjected to the following treatments: (i) unfixed and fixed spermatozoa were stained with fluorescein-labelled lectins; (ii) unfixed spermatozoa were incubated with lectins for determination of agglutination; and (iii) spermatozoa were incubated with detergent to remove the plasmalemma, the glycoproteins were separated on SDS PAGE and western blots were stained with biotinylated lectins. Many of the fluorescein isothiocyanate (FITC)-labelled lectins bound selectively to the sperm surface, and marked differences were found in lectin staining affinity between caput and cauda epididymal spermatozoa. Incubation of spermatozoa from the cauda epididymidis with neuraminidase reversed many of the differences in staining of the cauda epididymal spermatozoa, indicating masking of some terminal saccharides by sialic acid. Agglutination of spermatozoa from the caput epididymidis occurred after incubation with Concanavalin A (ConA) and soybean agglutinin (SBA), but agglutination was less extensive for spermatozoa from the cauda epididymidis. Western blot analysis indicated several ConA-positive bands in caput sperm extracts, but fewer positive bands in the cauda sperm extracts, whereas SBA stained four bands from caput but none from the cauda epididymal spermatozoa. These results demonstrate extensive glycosylation of the surface proteins of spermatozoa from the caput epididymidis and significant differences in spermatozoa from the cauda epididymidis. In general, the findings indicate similar glycosylation of the surface of marsupial spermatozoa to those from eutherian mammals despite marked differences in their morphology and early divergence of marsupials from eutherian mammals. It would appear that this situation differs markedly from that in sub-mammalian vertebrates. PMID- 11425342 TI - Activation of cumulus-free equine oocytes: effect of maturation medium, calcium ionophore concentration and duration of cycloheximide exposure. AB - Two different culture media (TCM-199 and follicular fluid), two activation treatments (10 and 50 micromol calcium ionophore l(-1)) and three culture periods with cycloheximide were evaluated to find effective culture conditions for activation of cumulus-free equine oocytes. Oocytes were collected by scraping the follicle walls of ovaries obtained from an abattoir. Oocytes with expanded cumuli were matured at 38.2 degrees C in a humidified atmosphere of 5% CO(2) in air, in either TCM-199 with 10% fetal bovine serum (FBS) and 5 microU FSH ml(-1), or in 100% follicular fluid derived from a preovulatory follicle 24 h after injection of hCG. After 40--42 h of in vitro maturation, oocytes were denuded by gentle pipetting in TCM-199 plus 10% FBS with hyaluronidase. Oocytes with intact cytoplasmic membranes (n = 398; 94% presumed metaphase II) were treated in protein-free PBS with 10 or 50 micromol calcium ionophore l(-1) for 5 min. After washing, the oocytes were cultured in TCM-199 containing 10% FBS and 10 microg cycloheximide ml(-1) for 6 h, in cycloheximide for 6 h and then in cycloheximide free medium for 18 h, or in cycloheximide for 24 h. The oocytes were fixed and evaluated by fluorescence microscopy. Oocytes with pronucleus I--II (dense to decondensing chromatin), pronucleus III--IV (decondensed chromatin) or progressing towards the first cleavage division were considered activated. The activation rate for oocytes matured in TCM-199 was significantly (P < 0.05) higher than for oocytes matured in follicular fluid (49% (99/204) versus 35% (60/171), respectively; P < 0.05). Culture with cycloheximide for 24 h resulted in a significantly higher rate of activation (67%, 74/111) than did the 6 h (33%, 44/136) or 6 h plus 18 h (32%, 41/128) treatments. The highest rate of activation (82%) was observed in oocytes matured in TCM-199, treated with 50 micromol calcium ionophore l(-1) and cultured with cycloheximide for 24 h. PMID- 11425344 TI - Editorial 1240 1240 Laurence H. Klotz EDITORIAL. PMID- 11425345 TI - Revision of species of Minerisporites, Azolla and associated plant microfossils from deposits of the Upper Palaeocene and Palaeocene/Eocene transition in the Netherlands, Belgium and the USA. AB - Species of the megaspore genus Minerisporites Potonie, megaspore apparatuses of species of the water fern Azolla Lamarck, and some associated organic-walled microfossils recovered from deposits of the Upper Palaeocene and Palaeocene/Eocene transition in the southern part of the Netherlands and neighbouring Belgium are redescribed on the basis of an examination of specimens under scanning and transmission electron microscopes. Originally studied about 40 years ago by S.J. Dijkstra, the re-examination has enabled emended diagnoses to be produced for six taxa: Minerisporites glossoferus (Dijkstra) Tschudy, M. mirabilis (Miner) Potonie, M. mirabilissimus (Dijkstra) Potonie, Azolla schopfii Dijkstra, A. teschiana Florschutz, and A. velus (Dijkstra) Jain and Hall. In addition, a revised description is provided for massulae of Salvinia Seguier that were originally thought to be megaspores and, hence, named by Dijkstra as Triletes? exiguus. The gross morphology and construction of the exospore of the species of Minerisporites are similar, but nevertheless sufficiently distinct for them to be maintained as separate taxa. Monolete microspores are preserved in hollows in the reticulate surface of some of the specimens of M. mirabilissimus. This is consistent with the presumed isoetalean affinity of Minerisporites. An apparent stratigraphic morphocline from M. glossoferus to M. mirabilis, suggested previously, is confirmed following our reassessment of their characteristics. The species of Azolla are all multi-floated, but they differ from each other in several ways, in particular with respect to the ultrastructure of the megaspore wall. They are also distinct from all other species that have been considered in sufficient detail for satisfactory comparisons to be made. The massulae of A. teschiana are described for the first time. The floats in A. velus are attached to the proximal part of the megaspore only by suprafilosal hairs. There are no maniculae. It is argued that these structures do not exist in either this or any other species of Azolla. Whole fertile fossil remains that demonstrate the nature and habitat of the living plants are known for all three genera. Hence, their reproductive structures can be used with confidence in palaeoenvironmental interpretation. The overall assemblage, which also includes a few specimens of the marsilealean megaspore Molaspora lobata (Dijkstra) Hall, undoubtedly reflects the presence of lowland aquatic and semi-aquatic vegetation. PMID- 11425343 TI - Biochemical, electron microscopic and immunohistological observations of cationic detergent-extracted cells: detection and improved preservation of microextensions and ultramicroextensions. AB - BACKGROUND: Filopodia, retraction fibers and microvilli, are fragile microextensions of the plasma membrane that are easily damaged by mechanical force during specimen preparation for microscopy. To preserve these structures for electron microscopy glutaraldehyde is generally used, but it often causes antigen masking. By contrast, formaldehyde is generally used for immunofluorescence light microscopy, but few studies have been concerned with the loss of microextensions. RESULTS: We demonstrate in biochemical experiments that cultured cells needed to be kept in 4% formaldehyde for at least 60 min at room temperature or for 20 min at 37 degrees C to irreversibly crosslink most of the polypeptides. Also, fragmentation of fragile microextensions was observed after Triton X-100 extraction depending on concentration and extent of crosslinking. We also report on a novel fixation procedure that includes the cationic detergent dodecyltrimethylammonium chloride (DOTMAC). Treatment of NIH3T3 cells with DOTMAC resulted in complete removal of membrane lipids and in good preservation of the cytoskeleton in microextensions as well as preservation of ultramicroextensions of <0.05 microm in diameter that have not been observed previously unless glutaraldehyde was used. Stress fibers and microextensions of DOTMAC-extracted cells were readily stained with anti-beta-actin antibodies, and antibodies to vinculin and moesin stained focal contacts and microextensions, respectively. CONCLUSIONS: Some microextensions were fragmented by the standard Triton X-100 permeabilization method. By contrast, DOTMAC completely extracted membrane lipids while maintaining the cytoskeleton of microextensions. Thus, DOTMAC treatment may provide a valuable new tool for the reliable visualization of previously undetectable or poorly detectable antigens while preserving the actin cytoskeleton of microextensions. PMID- 11425346 TI - An Early Cretaceous root-climbing epiphyte (Lindsaeaceae) and its significance for calibrating the diversification of polypodiaceous ferns. AB - The discovery of fossilised roots of a lindsaeoid fern within the trunk of the extinct tree fern Tempskya provides new fossil evidence for Lindsaeaceae in the Mesozic, as well as the first direct evidence of an ecological association between Tempskya and root-climbing epiphytes. Data were collected from permineralised Tempskya trunks from the Aspen Shale (Early Cretaceous, Albian), Wyoming (USA). The roots of the lindsaeoid fossil are clearly distinguishable from those of Tempskya and other living ferns based on a suite of distinctive anatomical features, which are described in detail. The foliage is unknown. The new fossil is interpreted as a root climber, and comparisons are made with similar living species in Lindsaea subgenus Odontoloma (Lindsaeaceae). The documentation of Lindsaeaceae in the Early Cretaceous adds to a small but growing body of data on the diversification of polypodiaceous ferns during this important period. These recent discoveries point to a much earlier (Early Cretaceous) crown group radiation of polypodiaceous ferns than previously suspected. PMID- 11425347 TI - Quantitative evaluation of microplankton palaeobiogeography in the Ordovician Early Silurian of the northern Trans European Suture Zone: implications for the timing of the Avalonia-Baltica collision. AB - Quantitative analysis of assemblage similarity among chitinozoan and acritarch associations recovered from various sedimentary sequences across the Trans European Suture Zone (TESZ; southern Baltic Sea and northern Germany region), permits evaluation of changes in microplankton palaeobiogeography during the Ordovician in the study area. The present data confirm strong palaeobiogeographic differences between the lower Ordovician of the Rugen area, and the coeval domains of the East European Platform (EEP), corroborating the idea that the subsurface of Rugen should be considered palaeogeographically as the eastern extension of Avalonia.Cluster analysis of chitinozoan assemblages from numerous wells in the Rugen area, and one well from the southern margin of the EEP indicates that chitinozoan bioprovincialism reached its maximum during the Llanvirn; during this period, the Rugen microplankton communities were clearly Gondwanan in character. Calculations using the coefficient of similarity support the conclusion of a high similarity between Llanvirn acritarch assemblages from the Rugen subsurface and from coeval Perigondwana localities (e.g. Tunisia). Since the early Caradoc, this Gondwanan affinity of the Rugen microfossils starts to lessen, and becomes negligible during the late Caradoc. During latest Caradoc early Ashgill through Llandovery times the chitinozoan assemblages from either side of the TESZ are undistinguishable. If palaeobiogeographical differentiation is primarily related to palaeolatitudinal distance, then the present data support closure of the Tornquist Ocean during late Caradoc-Ashgill times. The presence of reworked Llanvirn acritarchs of Perigondwanan affinity in middle Ashgill sedimentary sequences at the southern margin of the EEP, clearly shows that by this time erosion of an uplifted area was taking place. Accordingly, the closure of the Tornquist Ocean, and consequent Avalonia-Baltica collision must have taken place during the time interval between the middle Caradoc and the early middle Ashgill (Rawtheyan).New chitinozoan data from boreholes H 2 and K 5, offshore of Rugen Island, northern Germany, allow for precise biostratigraphic dating. In the pre-Devonian part of H 2, Siphonochitina formosa indicates an Abereiddian (Early Llanvirn) age, facilitating correlation with boreholes Binz 1/73 and Lohme 2/70 of Rugen. In borehole K 5, the occurrence of Belonechitina robusta and Tanuchitina bergstroemi permits attribution of the investigated pre-Carboniferous sequence to the middle-late Caradoc, and, possibly, early Ashgill. PMID- 11425348 TI - Orthopithonella collaris sp. nov., a new calcareous dinoflagellate cyst from the K/T boundary (Fish Clay, Stevns Klint/Denmark). AB - A new calcareous dinoflagellate cyst species, Orthopithonella collaris sp. nov., is described from the Cretaceous/Tertiary (K/T) boundary clay (Fish Clay) of Stevns Klint, Denmark, on the basis of SEM studies and light-microscopic analyses of thin sections of single specimens. The species has been found exclusively in the Fish Clay and as such may be a potential marker for the K/T boundary. Its pulse-like occurrence is thought to be due to the abrupt, relatively short-term ecological catastrophe associated with the K/T boundary event. PMID- 11425349 TI - Archaeopteris from the Upper Famennian of Belgium: heterospory, nomenclature, and palaeobiogeography. AB - New fertile specimens of Archaeopteris roemeriana from Upper (but not uppermost) Famennian strata of Belgium have been studied. Crowded sporangia are borne on non laminate, three times bifurcate fertile leaves helically arranged around the fertile ultimate axes. Vegetative leaves occur both proximal and distal to the fertile leaves. Heterospory is demonstrated with miospores belonging to the Geminospora-Aneurospora complex and macrospores most probably to Contagisporites genus. The established morphological traits allow us to substantiate a synonymy between A. roemeriana and A. halliana. The apparent dominance of A. halliana on the eastern side of the Acadian mountains during Late Famennian times is discussed. PMID- 11425350 TI - Inflorescence with tricolpate pollen grains from the Cenomanian of Tschulymo Yenisey Basin, West Siberia. AB - A new taxon Freyantha sibirica gen. et sp. nov. is erected for a delicate racemose inflorescence from the Cenomanian of the Tchulymo-Yenisey Basin, West Siberia. The inflorescence bears male flowers of partly connate stamens that produced tricolpate reticulate pollen grains. The flowers are subtended by calyptrate bracts. The taxonomically significant features of the general inflorescence morphology, prophylls, floral bracts, stamens and pollen grains are shared with different angiosperm groups, such as the Menispermaceae and Sargentodoxaceae. Prominent glands on the floral bracts indicate pollination by nectar-sucking insects. PMID- 11425352 TI - Variations in the presentation of primary intraocular lymphoma: case reports and a review. AB - Primary intraocular lymphoma is a distinct subset of primary non-Hodgkin's lymphoma of the central nervous system (CNS). Diagnosis can be difficult and is often delayed, as the clinical presentation can mimic a number of other ocular conditions. This report describes four different presentations of intraocular lymphoma and focuses on its modes of clinical presentation. Primary intraocular lymphoma can present with a wide variety of manifestations frequently mimicking diffuse uveitis that is refractory to corticosteroids. Subretinal pigment epithelium tumors may be seen. However, other presentations may include multiple deep white dots in the retina secondary to tumor infiltration; retinal infiltration, causing a necrotizing retinitis; or infiltration of the retinal vasculature, causing arterial or venous obstruction. Finally, optic nerve invasion may be seen. CNS lymphoma develops in the majority of patients before, in conjunction with, or after the development of eye disease. Intraocular lymphoma often has a fatal outcome, but recognition of its modes of presentation facilitates early diagnosis and treatment that may improve prognosis. PMID- 11425354 TI - Management of posterior capsule tears. AB - Any breach in the continuity of the posterior capsule is defined as a posterior capsule tear. Posterior capsule tears can be preexisting (congenital or traumatic), spontaneous, or intrasurgical. Preexisting/congenital posterior capsule tears have been related to an intrauterine insult. Posterior capsule tears due to trauma may occur as a consequence of direct mechanical impact due to perforation or blunt injury. Depending on the duration of time between the posterior capsular trauma and the cataract surgery, these posterior capsule tears can have different features. Intrasurgical posterior capsule tears are the most common and can occur during any stage of cataract surgery. Also, they may be planned in the form of primary posterior capsulorhexis. The conventional management consists of prevention of mixture of cortical matter with vitreous, dry aspiration, and anterior vitrectomy, if required. In addition, during phacoemulsification low flow rate, high vacuum, and low ultrasound are advocated if a posterior capsule tear occurs. Dislocated nucleus or nuclear fragments require vitrectomy and the use of perfluorocarbon liquids. In the presence of a posterior capsule tear, the IOL can be placed in the sulcus, if the capsular rim is available, or in the bag, if the tear is small. Scleral fixated posterior chamber lenses and anterior chamber IOLs can be implanted when the posterior capsule tear is large. PMID- 11425355 TI - True, true and related? AB - Several chorioretinal lesions have been observed that are associated with bone marrow transplantation (BMT), such as cotton-wool spots, macular stars, ischemic changes due to microangiopathy, "BMT retinopathy" and choroidal infiltration. Central serous retinopathy (CSR) has rarely been described in the BMT setting. We present a patient who underwent allogeneic BMT and subsequently developed severe chronic graft versus host disease (CGvHD) complicated with CSR. PMID- 11425356 TI - The Draize eye test. AB - Hundreds of substances are used daily that can damage eyesight. People's eyes are open to accidental or intentional exposure during the production, transportation, use, and disposal of chemical preparations. Ensuring the safety of consumer products was born during the mid twentieth century in the aftermath of chemical warfare research, and was motivated by the hazards of unsafe cosmetics. Justified by an exigency for public protection, the Draize eye test became a governmentally endorsed method to evaluate the safety of materials meant for use in or around the eyes. The test involves a standardized protocol for instilling agents onto the cornea and conjunctiva of laboratory animals. A sum of ordinal-scale items of the outer eye gives an index of ocular morbidity. Advances in ocular toxicology are challenging the validity, precision, relevance, and need of the Draize eye test. Preclinical product-safety tests with rabbits and other mammals also raise ethical concerns of animal wellbeing. Some use the Draize test as a rallying point for how animals are treated in science and industry. A battery of cellular systems and computer models aim to reduce and ultimately to replace whole-animal testing. Molecular measures of ocular toxicity may eventually allow comprehensive screening in humans. The Draize eye test was created and refined for humanitarian reasons and has assuredly prevented harm. Its destiny is to be progressively supplanted as in vitro and clinical alternatives emerge for assessing irritancy of the ocular surface. PMID- 11425357 TI - Alzheimer's disease: a review for the ophthalmologist. AB - Alzheimer's disease is the most prevalent dementia. Definitive diagnosis is made only at autopsy, although "probable" diagnoses are made using clinical criteria. Patients with Alzheimer's disease demonstrate severe deficits in memory with cortical features of language difficulty and visuomotor spatial deficits. They also may present with psychotic symptoms of delusions and hallucinations, and personality and behavioral changes. Advancing age, genetics, and environmental risk factors are important in the development of Alzheimer's disease. Visual abnormalities have been described in Alzheimer's disease and may be related to the development of visual hallucinations. Although palliative treatments exist for the cognitive loss and behavioral symptoms, future treatments will focus on both delay of onset and slowing of progression of the disease. Continued research is needed to further understand this devastating disorder, which may in turn lead to more successful treatments. PMID- 11425358 TI - A. Franceschetti--great ophthalmologist and humanitarian. AB - A. Franceschetti is well known for his work in ocular genetics; however, his role in creating a department that served as a haven for ophthalmologists during World War II is less well known. Franceschetti was an enthusiastic and multitalented researcher whose contributions to ophthalmology are far-reaching. PMID- 11425359 TI - Visual function assessment questionnaires. AB - With increased emphasis on functional outcomes in ophthalmology, third-party health care payers and research funding agencies have turned their attention to the development and use of visual function questionnaires. Since 1980, more than a dozen such self-report visual function questionnaires have been developed. All of these instruments include items that ask about specific daily activities; patients must respond with a rating that represents the level of difficulty that they experience with the activity described. This article reviews all of the known instruments, with special attention paid to their validity and reliability. Most validation studies have reported high response consistency across items and significant correlations of instrument scores with visual impairment measures. Only two studies have measured test-retest reliability. The developers of visual function questionnaires typically divide the items into several different subscales, suggesting that different variables are being measured. Although the items are very similar for the different instruments, there is little agreement among instruments on the definition of subscales. All instruments are scored as the average of the ordinal patient ratings across items for each subscale and/or for the total instrument. Measurement issues underlying the scoring of ordinal patient ratings are discussed. It is argued that unless the instruments can be converted to interval scales, the averaging of patient ratings does not yield true measurements. The three visual function questionnaires that were calibrated with a statistical item response model, which estimates interval scales, are reviewed. It is concluded that future research and development should devote additional attention to the measurement properties of functional assessment instruments. PMID- 11425360 TI - Why rank countries by health performance? PMID- 11425361 TI - Does breastfeeding really affect mortality among HIV-1 infected women? PMID- 11425362 TI - Unsatisfactory redefinition of myocardial infarction. PMID- 11425363 TI - Prospects for the prevention of haemolytic-uraemic syndrome. PMID- 11425364 TI - The new pan-Asian paan problem. PMID- 11425365 TI - Leprosy and the genome--not yet a burnt-out case. PMID- 11425366 TI - Behavioural problems in children who weigh 1000 g or less at birth in four countries. AB - BACKGROUND: The increased survival chances of extremely low-birthweight (ELBW) infants (weighing <1000 g at birth) has led to concern about their behavioural outcome in childhood. In reports from several countries with different assessments at various ages, investigators have noted a higher frequency of behavioural problems in such infants, but cross-cultural comparisons are lacking. Our aim was to compare behavioural problems in ELBW children of similar ages from four countries. METHODS: We prospectively studied 408 ELBW children aged 8-10 years, whose parents completed the child behaviour checklist. The children came from the Netherlands, Germany, Canada, and USA. The checklist provides a total problem score consisting of eight narrow-band scales. Of these, two (aggressive and delinquent behaviour) give a broad-band externalising score, three (anxious, somatic, and withdrawn behaviour) give a broad-band internalising score, and three (social, thought, and attention problems) indicate difficulties fitting neither broad-band dimension. For each cohort we analysed scores in ELBW children and those in normal- birthweight controls (two cohorts) or national normative controls (two cohorts). Across countries, we assessed deviations of the ELBW children from normative or control groups. FINDINGS: ELBW children had higher total problem scores than normative or control children, but this increase was only significant in European countries. Narrow-band scores were raised only for the social, thought, and attention difficulty scales, which were 0.5-1.2 SD higher in ELBW children than in others. Except for the increase in internalising scores recorded for one cohort, ELBW children did not differ from normative or control children on internalising or externalising scales. INTERPRETATION: Despite cultural differences, types of behavioural problems seen in ELBW children were very similar in the four countries. This finding suggests that biological mechanisms contribute to behavioural problems of ELBW children. PMID- 11425367 TI - Application of an immunodiagnostic method for improving preoperative diagnosis of nodular thyroid lesions. AB - BACKGROUND: Thyroid cancer is the most common endocrine malignant disease, but preoperative diagnosis remains a challenge. Fine-needle aspiration cytology has greatly improved the clinical management of thyroid nodules, but the preoperative characterisation of follicular lesions is very difficult. Many patients are thus referred to surgery more for diagnosis than for therapeutic necessity. We undertook an international multicentre study to assess the usefulness of immunohistocytochemical staining for two potential markers of malignant thyrocytes. METHODS: Expression of galectin-3 and CD44v6 was tested on 1009 thyroid lesions (tissue specimens and cytological cell-blocks) and 226 fresh cytological samples obtained preoperatively by ultrasound-guided fine-needle aspiration of thyroid nodules (prospective analysis). The test used monoclonal antibodies specific for CD44v6 and galectin-3, the indirect avidin-biotin complex immunoperoxidase method, and 3-amino-9-ethyl-carbazole as substrate. FINDINGS: The sensitivity, specificity, positive predictive value, and diagnostic accuracy of this test method (for coexpression of the two markers) in the prospective analysis were 88%, 98%, 91%, and 97%, respectively. The sensitivity and specificity of galectin-3 immunodetection alone in discriminating benign from malignant thyroid lesions were more than 99% and 98% respectively, and the positive predictive value and diagnostic accuracy were 92% and 99%. INTERPRETATION: The integration of galectin-3 immunostaining with conventional cytomorphological and clinical diagnostic procedures represents a sensitive and reliable diagnostic approach for preoperative identification of thyroid carcinomas. This test method improves the diagnostic accuracy of conventional cytology and provides the molecular basis for a new nosological assignation of the not yet classified thyroid neoplasms of indeterminate malignant behaviour. PMID- 11425369 TI - Effect of breastfeeding on mortality among HIV-1 infected women: a randomised trial. AB - BACKGROUND: We have completed a randomised clinical trial of breastfeeding and formula feeding to identify the frequency of breastmilk transmission of HIV-1 to infants. However, we also analysed data from this trial to examine the effect of breastfeeding on maternal death rates during 2 years after delivery. We report our findings from this secondary analysis. METHODS: Pregnant women attending four Nairobi city council clinics were offered HIVtests. At about 32 weeks' gestation, 425 HIV-1 seropositive women were randomly allocated to either breastfeed or formula feed their infants. After delivery, mother-infant pairs were followed up monthly during the first year and quarterly during the second year until death, or 2 years after delivery, or end of study. FINDINGS: Mortality among mothers was higher in the breastfeeding group than in the formula group (18 vs 6 deaths, log rank test, p=0.009). The cumulative probability of maternal death at 24 months after delivery was 10.5% in the breastfeeding group and 3.8% in the formula group (p=0.02). The relative risk of death for breastfeeding mothers versus formula feeding mothers was 3.2 (95% CI 1.3-8.1, p=0.01). The attributable risk of maternal death due to breastfeeding was 69%. There was an association between maternal death and subsequent infant death, even after infant HIV-1 infection status was controlled for (relative risk 7.9, 95% CI 3.3-18.6, p<0.001). INTERPRETATION: Our findings suggest that breastfeeding by HIV-1 infected women might result in adverse outcomes for both mother and infant. PMID- 11425370 TI - Nocturnal hypoxaemia and central-nervous-system events in sickle-cell disease. AB - BACKGROUND: Central-nervous-system (CNS) events, including strokes, transient ischaemic attacks, and seizures are common in sickle-cell disease. Stroke can be predicted by high velocities in the internal-carotid or middle-cerebral arteries on transcranial doppler ultrasonography. We tested the hypothesis that nocturnal hypoxaemia can predict CNS events better than clinical or haematological features, or transcranial doppler sonography. METHODS: We screened 95 hospital based patients with sickle-cell disease (median age 7.7 years [range 1.0-23.1]), but without previous stroke, with transcranial doppler and overnight pulse oximetry. Follow-up continued for a median of 6.01 (0.11-8.54) years. FINDINGS: 19 patients had CNS events (six ischaemic and one haemorrhagic stroke, eight transient ischaemic attacks, and four seizures). Mean overnight oxygen saturation ([SaO(2)] hazard ratio 0.82 per 1% increase [95% CI 0.71-0.93]; p=0.003) and higher internal-carotid or middle-cerebral artery velocity (1.02 for every increase of 1 cm/s [1.004-1.03]; p=0.009) were independently associated with time to CNS event. After accounting for mean SaO(2), artery velocity, and haemoglobinopathy, high haemoglobin concentration was also associated with an increased risk of CNS event (1.7 per g/dL, [1.18-2.43]; p=0.004). Dips suggestive of obstructive sleep apnoea did not predict CNS events, and adenotonsillectomy seemed to have no effect, although the CI were wide and clinically important effects cannot be excluded. INTERPRETATION: Screening for, and appropriate management of, nocturnal hypoxaemia might be a safe and effective alternative to prophylactic blood transfusion for primary prevention of CNS events in sickle cell disease. PMID- 11425371 TI - Renal function, cytogenetic measurements, and sexual development in adolescents in relation to environmental pollutants: a feasibility study of biomarkers. AB - BACKGROUND: Human exposure to chemicals is normally monitored by measurement of environmental pollutants in external media. We investigated whether biomarkers in adolescents can show exposure to, and health effects of, common environmental pollutants. METHODS: We recruited 200 17-year-old adolescents (120 girls) from a rural control area and from two suburbs polluted by a lead smelter and two waste incinerators. We measured biomarkers of exposure and of effect in blood and urine samples, and obtained questionnaire data. School doctors measured testicular volume and staged sexual maturation. FINDINGS: Internal exposure was mostly within current standards. Concentrations of lead and cadmium in blood, PCBs (polychlorinated biphenyls) and dioxin-like compounds in serum samples, and metabolites of VOCs (volatile organic compounds) in urine were higher in one or both suburbs than in the control area. Children who lived near the waste incinerators matured sexually at an older age than others, and testicular volume was smaller in boys from the suburbs than in controls. Biomarkers of glomerular or tubular renal dysfunction in individuals were positively correlated with blood lead. Biomarkers of DNA damage were positively correlated with urinary metabolites of PAHs (polycyclic aromatic hydrocarbons) and VOCs. Interpretation Biomarkers can be used to detect environmental exposure to pollutants and measure their biological effects before overt disease develops. Our findings suggest that current environmental standards are insufficient to avoid measurable biological effects. PMID- 11425372 TI - A diabetic breast lump. PMID- 11425373 TI - World Health Report 2000: inequality index and socioeconomic inequalities in mortality. AB - Monitoring of inequality in health has become an increasingly important task of development agencies. We compared the inequality index as published in the World Health Report 2000 with available evidence on socioeconomic inequalities in mortality in 15 industrialised and 43 less-developed countries. We found that the World Health Report index does not correspond with international variations in the size of socioeconomic inequalities in mortality. These findings indicate that the index should not be interpreted as a reflection of socioeconomic inequalities in health, nor should the index be used to replace the indices developed to monitor socioeconomic inequalities in health. PMID- 11425374 TI - Use of pentoxifylline in membranous nephropathy. AB - Urinary TNF-alpha excretion correlates with proteinuria in patients with membranous nephropathy (MGN). Pentoxifylline suppresses or reduces the production of TNF-alpha. Between April, 1999 and August, 2000, we did a single-centre, prospective, pilot study to assess the effects of pentoxifylline (1200 mg/day) on proteinuria in patients with idiopathic MGN. Ten patients were included and treated for 6 months. Pentoxifylline significantly decreased proteinuria from 11 g/day [range 4.6-27] to 1.8 (0-10.9); p=0.001). Pentoxifylline may be a safe and effective adjunct to steroids and immunosuppressants in patients with MGN. PMID- 11425375 TI - 25-hydroxyvitamin D-1alpha-hydroxylase in normal and malignant colon tissue. AB - Vitamin D affects calcium metabolism and prevents proliferation of colon cells in vitro. In human beings the main circulating form of vitamin D is 25 hydroxyvitamin D; to regulate calcium homoeostasis, this form must be converted to 1alpha, 25-dihydroxyvitamin D by 1alpha-hydroxylation in the kidney with 25 hydroxyvitamin D-1alpha-hydroxylase. Cultured transformed colon cancer cells can convert 25-hydroxyvitamin D(3) to 1alpha,25-dihydroxyvitamin D(3). We identified messenger RNA (mRNA) for 25-hydroxyvitamin D-1alpha-hydroxylase in normal colon tissue and in malignant and adjacent normal colon tissue. These findings support the notion that vitamin D might have a role in cell growth regulation and cancer protection, and might be the explanation for why the risk of dying from colorectal cancer is highest in areas with the least amount of sunlight. PMID- 11425376 TI - In-vivo transfer of mecA DNA to Staphylococcus aureus [corrected]. AB - Staphylococcus aureus is thought to have acquired mecA DNA by horizontal transfer. DNA fingerprints made by restriction nucleases that cut certain sequences of DNA are used to compare complete genomes or particular genes between bacteria. We isolated an epidemic mecA(-) meticillin-susceptible S aureus genotype and, subsequently, a rare isogeneic mecA(+) meticillin-resistant S aureus (MRSA) genotype from a neonate who had never been in contact with MRSA. This MRSA contained mecA DNA that was identical to that in a coagulase-negative staphylococcal strain isolated from this patient, but different from other MRSA genotypes. We believe that this MRSA was formed in vivo by horizontal transfer of the mecA DNA between two staphylococcal species. PMID- 11425377 TI - Trachoma and vitamin A deficiency. AB - Both vitamin A deficiency and trachoma are important causes of preventable ocular diseases. We examined 10 559 children in Mali for clinical signs of trachoma and of xerophthalmia. In five regions, the mean prevalence of trachoma was 39.3% (95% CI: 37.4-41.2). Clinical xerophthalmia was present among 1.89% of children (1.59 2.25). The diagnosis of xerophthalmia appeared to be associated with active trachoma (OR=2.04 [1.52-2.74]). This association remained after controlling for socio-economic status. PMID- 11425378 TI - Online database reveals researchers' industry ties. PMID- 11425379 TI - Automatic external defibrillation is safe in small children. PMID- 11425380 TI - BRCA1 has a pivotal role in repairing DNA. PMID- 11425381 TI - Crohn's gene identified. PMID- 11425382 TI - Proteolysis problems implicated in neurodegenerative diseases. PMID- 11425383 TI - Raised endothelial microparticles an early marker for multiple sclerosis? PMID- 11425384 TI - Tackling venomous snake bites worldwide. PMID- 11425385 TI - Buenos Aires new twist to Argentina's health-care crisis. PMID- 11425386 TI - Spain's government puts domestic violence protection plan as top priority. PMID- 11425387 TI - India tries to improve national system for handling clinical trials. PMID- 11425388 TI - US state given go ahead to reduce drug prices for uninsured people. PMID- 11425389 TI - WHO faces funding squeeze as richer countries make a stand. PMID- 11425390 TI - North Korean government admits that health of children is very poor. PMID- 11425391 TI - UK scientists review use of genetically modified animals in research. PMID- 11425392 TI - Healthy life expectancy in 191 countries, 1999. AB - We describe here the methods used to produce the first estimates of healthy life expectancy (DALE) for 191 countries in 1999. These were based on estimates of the incidence, prevalence, and disability distributions for 109 disease and injury causes by age group, sex, and region of the world, and an analysis of 60 representative health surveys across the world. We used Sullivan's method to compute healthy life expectancy for men and women in each WHO member country. Japan had the highest average healthy life expectancy of 74.5 years at birth in 1999. The bottom ten countries are all in sub-Saharan Africa, where the HIV-AIDS epidemic is most prevalent, resulting in DALE at birth of less than 35 years. Years of healthy life lost due to disability represent 18% of total life expectancy in the bottom countries, and decreases to around 8% in the countries with the highest healthy life expectancies. Globally, the male-female gap is lower for DALE than for total life expectancy. Healthy life expectancy increases across countries at a faster rate than total life expectancy, suggesting that reductions in mortality are accompanied by reductions in disability. Although women live longer, they spend a greater amount of time with disability. As average levels of health expenditure per capita increase, healthy life expectancy increases at a greater rate than total life expectancy. PMID- 11425393 TI - Uses of error: the wrong research costs lives. PMID- 11425394 TI - Methodological concerns and recommendations on policy consequences of the World Health Report 2000. PMID- 11425395 TI - World Health Report 2000: a step towards evidence-based health policy. PMID- 11425396 TI - World Health Report 2000: responses to Murray and Frenk. PMID- 11425403 TI - Cancer toll falling in the USA. PMID- 11425404 TI - GUSTO V: the bottom line of fibrinolytic reperfusion therapy. PMID- 11425405 TI - Glycoprotein IIb/IIIa receptor blockers in acute coronary syndromes: Gusto IV ACS. PMID- 11425406 TI - Stopping smoking: a hazard for people with a history of major depression? PMID- 11425407 TI - Endogenous opioids, placebo response, and pain. PMID- 11425408 TI - Crohn's disease: genetic susceptibility, bacteria, and innate immunity. PMID- 11425409 TI - Too much, too little, or none at all: dealing with substandard and fake drugs. PMID- 11425410 TI - Reperfusion therapy for acute myocardial infarction with fibrinolytic therapy or combination reduced fibrinolytic therapy and platelet glycoprotein IIb/IIIa inhibition: the GUSTO V randomised trial. AB - BACKGROUND: Plasminogen activator therapy for acute myocardial infarction is limited by lack of achievement of early, complete, and sustained reperfusion in a substantial proportion of patients. Many phase II trials have supported the potential of combined fibrinolytic therapy and platelet glycoprotein IIb/IIIa inhibition for improving reperfusion. We did a randomised, open-label trial to compare the effect of reteplase alone with reteplase plus abciximab in patients with acute myocardial infarction. METHODS: 16588 patients in the first 6 h of evolving ST-segment elevation myocardial infarction were randomly assigned standard-dose reteplase (n=8260) or half-dose reteplase and full-dose abciximab (n=8328). The primary endpoint was 30-day mortality, and secondary endpoints included various complications of myocardial infarction. Analysis was by intention to treat. FINDINGS: At 30 days, 488 (5.9%) of patients in the reteplase group had died, compared with 468 (5.6%) in the combined reteplase and abciximab group (odds ratio 0.95 [95% CI 0.83-1.08], p=0.43). There were fewer deaths or non-fatal reinfarctions with the combination than with reteplase alone, and there was less need for urgent revascularisation and fewer major non-fatal ischaemic complications of acute myocardial infarction. On the other hand, there were more non-intracranial bleeding complications in the combination group. The rates of intracranial haemorrhage and non-fatal disabling stroke were similar. INTERPRETATION: Although combined reteplase and abciximab was not superior to standard reteplase, the 0.3% absolute (5% relative) decrease in 30-day mortality fulfilled the criteria of non-inferiority. Combination therapy led to a consistent reduction in key secondary complications of myocardial infarction including reinfarction, which was partly counterbalanced by increased non intracranial bleeding complications. PMID- 11425411 TI - Effect of glycoprotein IIb/IIIa receptor blocker abciximab on outcome in patients with acute coronary syndromes without early coronary revascularisation: the GUSTO IV-ACS randomised trial. AB - BACKGROUND: Glycoprotein IIb/IIIa blockers reduce procedure-related thrombotic complications of percutaneous coronary intervention, and the risk of death and myocardial infarction in patients with acute coronary syndromes. The effect on risk of death and myocardial infarction is particularly apparent in patients undergoing early percutaneous coronary interventions. We did a randomised, multicentre trial to study the effect of the glycoprotein IIb/IIIa blocker abciximab on patients with acute coronary syndromes who were not undergoing early revascularisation. METHODS: We enrolled 7800 patients who were admitted to hospital with chest pain and either ST-segment depression or raised troponin T or I concentrations. 2598 were randomly assigned placebo, 2590 an abciximab bolus and 24 h infusion, and 2612 an abciximab bolus and 48 h infusion; all patients received aspirin and either unfractionated or low-molecular-weight heparin. The primary endpoint was death or myocardial infarction at 30 days after randomisation. Analysis was by intention to treat. FINDINGS: There were no drop outs. 209 (8.0%) patients on placebo, 212 (8.2%) on 24 h abciximab, and 238 (9.1%) on 48 h abciximab died or had a myocardial infarction before day 30 (odds ratio 1.0 [95% CI 0.83-1.24], for difference between placebo and 24 h abciximab, and 1.1 [0.94-1.39] for difference between placebo and 48 h abciximab). The lack of benefit from treatment with abciximab was consistent in most subgroups investigated; in particular, no benefit was seen in patients with raised cardiac troponin T or I concentrations at enrolment, although these patients did have a strongly increased risk of subsequent events. Bleeding rates were low, but increased with abciximab, particularly when continued for 48 h. Additionally, thrombocytopenia was more frequent with abciximab than with placebo. INTERPRETATIONS: Although the explanations for our findings are unclear, this study indicates that abciximab is not beneficial as first-line medical treatment in patients admitted with acute coronary syndromes. PMID- 11425413 TI - Association between insertion mutation in NOD2 gene and Crohn's disease in German and British populations. AB - Background Genetic predisposition to inflammatory bowel disease (IBD) has been shown by epidemiological and linkage studies. Genetic linkage of IBD to chromosome 16 has been previously observed and replicated in independent populations. The recently identified NOD2 gene is a good positional and functional candidate gene since it is located in the region of linkage on chromosome 16q12, and activates nuclear factor (NF) kappaB in response to bacterial lipopolysaccharides. Methods We sequenced the coding region of the NOD2 gene and genotyped an insertion polymorphism affecting the leucine-rich region of the protein product in 512 individuals with IBD from 309 German or British families, 369 German trios (ie, German patients with sporadic IBD and their unaffected parents), and 272 normal controls. We then tested for association with Crohn's disease and ulcerative colitis. Findings Family-based association analyses were consistently positive in 95 British and 99 German affected sibling pairs with Crohn's disease (combined p<0.0001); the association was confirmed in the 304 German trios with Crohn's disease. No association was seen in the 115 sibling pairs and 65 trios with ulcerative colitis. The genotype-specific disease risks conferred by heterozygous and homozygous mutant genotypes were 2.6 (95% CI 1.5-4.5) and 42.1 (4.3-infinity), respectively. Interpretation The insertion mutation in the NOD2 gene confers a substantially increased susceptibility to Crohn's disease but not to ulcerative colitis. PMID- 11425414 TI - Smoking cessation and the course of major depression: a follow-up study. AB - BACKGROUND: Smokers with a history of major depression who attempt to stop smoking have a higher risk of failure than non-depressed smokers. Anecdotal and post-hoc data suggest that those who successfully abstain are at increased risk of depression compared with individuals who continue to smoke. However, these studies confound effects of abstinence and history of depression. We aimed to assess whether there is an increased risk of depression and for how long that increase lasts. METHODS: We enrolled 100 smokers (>1 pack per day) with a history of major depression, but who were currently free from major depression and had not been on antidepressant medicine for at least 6 months, in a 2-month smoking cessation trial. The primary outcome was recurrence of major depression, which we assessed by structured clinical interviews 3 and 6 months after the end of treatment. We verified smoking status by serum-sample cotinine concentrations. FINDINGS: 76 participants (42 successful abstainers, 34 smokers) were followed up. 13 abstainers and two smokers had an episode of major depression (odds ratio 7.17 [95% CI 1.5-34.5]; Kaplan-Meier survival curve, log-rank statistic 9.11 [p=003]). Risk of major depression was similar between the first and second 3 months of follow-up. INTERPRETATIONS: Smokers with a history of depression who abstain from smoking are at significantly increased risk of developing a new episode of major depression. This risk remains high for at least 6 months. PMID- 11425415 TI - Pharmacopoeial quality of drugs supplied by Nigerian pharmacies. AB - BACKGROUND: The quality of medicines available in some less-developed countries is inadequate in terms of content of active ingredient. Reasons for the poor quality of drugs include widespread counterfeiting of medicines in less-developed countries, excessive decomposition of active ingredient as a result of high temperature and humidity, and poor quality assurance during the manufacture of medicinal products. Our aim was to investigate the quality of different drugs obtained from retail pharmacies in two urban areas of Nigeria, and, in instances of poor quality, to ascertain the reason why. METHODS: We randomly collected 581 samples of 27 different drugs from 35 pharmacies in Lagos and Abuja in Nigeria. We analysed the medicines for drug content by validated chromatographic methods, and compared our results with pharmacopoeial requirements. FINDINGS: 279 (48%) samples did not comply with set pharmacopoeial limits, and this proportion was uniform for the various types of drugs tested. Although some preparations contained no active ingredient, most had amounts just outside the pharmacopoeial limits. We identified samples with both too much and too little active drug content. INTERPRETATION: The most probable cause of the poor quality of drugs is absence of adequate quality assurance during manufacture. Substandard drugs sold in the pharmacies of less-developed countries could contribute to global microbial resistance and therapeutic failure of infectious diseases. PMID- 11425416 TI - Safety and antigenicity of non-adjuvanted and MF59-adjuvanted influenza A/Duck/Singapore/97 (H5N3) vaccine: a randomised trial of two potential vaccines against H5N1 influenza. AB - BACKGROUND: In 1997, pathogenic avian influenza A/Hong Kong/97 (H5N1) viruses emerged as a pandemic threat to human beings. A non-pathogenic variant, influenza A/Duck/Singapore/97 (H5N3), was identified as a leading vaccine candidate. We did an observer-blind, phase I, randomised trial in healthy volunteers to assess safety, tolerability, and antigenicity of MF59-adjuvanted and non-adjuvanted vaccines. METHODS: 32 participants were randomly assigned MF59, and 33 non adjuvanted vaccine. Two doses were given 3 weeks apart, of 7.5, 15, or 30 microg haemagglutinin surface-antigen influenza A H5N3 vaccine. Antibody responses were measured by haemagglutination inhibition, microneutralisation, and single radial haemolysis (SRH). The primary outcome was geometric mean antibody titre 21 days after vaccination. FINDINGS: The A/Duck/SIngapore vaccines were safe and well tolerated. Antibody response to non-adjuvanted vaccine was poor, the best response occurring after two 30 microgram doses: one, four, four, and one person of eleven seroconverted by haemagglutination inhibition, microneutralisation, H5N3 SRH, and H5N1 SRH, respectively. The geometric mean titres of antibody, and seroconversion rates, were significantly higher after MF59 adjuvanted vaccine. Two 7.5 microg doses of MF59 adjuvanted vaccine gave the highest seroconversion rates: haemagglutination inhibition, six of ten; microneutralisation, eight of ten; H5N3 SRH, ten of ten; H5N1 SRH, nine of ten. Geometric mean titre of antibody to the pathogenic virus, A/Hong Kong/489/97 (H5N1), was about half that to A/Duck/Singapore virus. INTERPRETATION: Non-adjuvanted A/Duck/Singapore/97 (H5N3) vaccines are poorly immunogenic and doses of 7.5-30 microg haemagglutinin alone are unlikely to give protection from A/Hong Kong/97 (H5N1) virus. Addition of MF59 to A/Duck/Singapore/97 vaccines boost the antibody response to protection levels. Our findings have implications for development and assessment of vaccines for future pandemics. PMID- 11425417 TI - Coma in a young anorexic woman. PMID- 11425418 TI - Genetic determinants of jaundice and gallstones in haemoglobin E beta thalassaemia. AB - Chronic hyperbilirubinaemia, gallstone formation, and gall bladder disease are unusually common in people with haemoglobin E beta thalassaemia in Sri Lanka. To determine whether this has a genetic basis we compared the bilirubin levels and frequency of gallstones in patients with different alleles of the UGT*1 gene. There was a significantly higher bilirubin level in those with the 7/7 genotypes compared with 6/6 and 6/7 genotype (p=0.032 and 0.0015 respectively), who also appeared more prone to gallstone formation. These results suggest that the UGT*1 genotpe is of importance in the genesis of gallstones in this population of patients. PMID- 11425419 TI - Acute tubulointerstitial nephritis associated with the selective COX-2 enzyme inhibitor, rofecoxib. AB - The nephrotoxic effect of COX-2 selective inhibitors has not yet been established. We report a case of reversible acute renal failure due to acute tubulointerstitial nephritis, confirmed by histology of a renal biopsy sample, associated with taking rofecoxib, a selective cyclo-oxygenase-2 (COX-2) inhibitor. PMID- 11425420 TI - In-utero treatment of fetal bladder-outlet obstruction by a ureterocele. AB - Fetal bladder-outlet obstruction by ureteroceles is usually treated at birth. However, such obstruction may be detrimental to the health of the fetus and so in utero correction is preferable. We describe the successful cytoscopy guided laser incision, with no complications, of a uterocele that was causing bladder-outlet obstruction in a fetus of 19 weeks and 6 days gestation. PMID- 11425421 TI - Fake artesunate in southeast Asia. AB - Artesunate is a key antimalarial drug in the treatment of multidrug-resistant Plasmodium falciparum malaria in southeast Asia. We investigated the distribution of counterfeit artesunate tablets by use of the validated, simple, and inexpensive Fast Red TR dye technique. We also aimed to identify distinguishing characteristics of the fake drugs. Of 104 shop-bought "artesunate" samples from Cambodia, Laos, Myanmar (Burma), Thailand, and Vietnam, 38% did not contain artesunate. Characteristics such as cost and physical appearance of the tablets and packaging reliably predicted authenticity. The illicit trade in counterfeit antimalarials is a great threat to the lives of patients with malaria. The dye test will assist national malaria control authorities in urgently needed campaigns to stop this murderous trade. PMID- 11425422 TI - Winter seasonality and rotavirus diarrhoea in adults. AB - We investigated the aetiological role of group A rotavirus in adults with acute diarrhoea in a 4-year prospective study. Of 683 patients with acute diarrhoea, 97 (14%) shed rotavirus as a sole agent, whereas six (5%) of 115 patients without diarrhoea shed rotavirus. Half of patients with rotavirus diarrhoea required admission to hospital. Unlike rotavirus diarrhoea in children, the occurrence of rotavirus-positive cases did not show a significant winter seasonality. Rotavirus infection should be included in the differential diagnosis of diarrhoeal diseases in adults. PMID- 11425423 TI - US paediatricians advised to ask about sexual assault. PMID- 11425425 TI - Air pollution can trigger a myocardial infarction. PMID- 11425426 TI - New technique may lead to a test for abnormal prion protein. PMID- 11425427 TI - Cell-to-cell interactions may hold key to better bone marrow transplants. PMID- 11425428 TI - Carpal tunnel syndrome study stirs controversy. PMID- 11425429 TI - Where should Labour's second term lead in health? PMID- 11425430 TI - Labour returns to power and a tough health agenda. PMID- 11425431 TI - Israeli psychiatrists propose new law for forced incarceration of patients. PMID- 11425432 TI - European Parliament takes first steps towards a regional food authority. PMID- 11425433 TI - Uganda launches HIV/AIDS treatment and training centre for Africa. PMID- 11425434 TI - European Union to fund mutant mouse archive for genomics research. PMID- 11425435 TI - Croatia starts to involve patients in medical decision-making. PMID- 11425436 TI - US smoker awarded record-breaking compensation. PMID- 11425437 TI - Advances in transplantation tolerance. AB - Immunosuppressive drugs developed in the past two decades have improved the short term survival of organ allografts, but tolerance has not been achieved and almost all transplant recipients continue to require drugs throughout life. Graft rejection arises from the cognate interaction of T cells with antigen-presenting cells, the recognition of alloantigen through the T-cell receptor, and the delivery of accessory stimulation signals. Once activated by the specific antigen, replicating T cells die if they are re-exposed to the same antigen. Since depletion of antigen-activated T cells is one critical mechanism of transplantation tolerance, drugs such as ciclosporin that interfere with activation-induced T-cell death could inhibit tolerance, whereas drugs such as mycophenolate mofetil, that induce the death of activated T cells, could facilitate tolerance. Other tolerance mechanisms depend on inactivation rather than elimination of allograft reactive T cells. When antigen recognition occurs without costimulation through the CD28 and CD154 accessory receptors, or in absence of cell division, T cells become unresponsive. Thus, inhibitors of CD28 and CD154, and inhibition of T-cell division by rapamycin promotes transplantation tolerance. PMID- 11425438 TI - Augmentation of nitric oxide to treat detrusor-external sphincter dyssynergia in spinal cord injury. AB - Detrusor-external sphincter dyssynergia (DSD) is a common cause of bladder outlet obstruction in men with spinal cord injuries, which if left untreated leads ultimately to renal failure. External sphincterotomy is currently the main treatment for DSD. However, obstruction persists in a substantial proportion of cases after this procedure. There is no effective drug treatment for DSD. Nitric oxide is an inhibitory neurotransmitter synthesised by nitric oxide synthase. Both animal and human studies suggest that nitric oxide mediates urethral sphincter relaxation. Nitric-oxide-synthase staining neurons have been identified in very high density in the urethral sphincters of a variety of animals and in human beings. Relaxation of the urethral sphincter is abolished by inhibitors of nitric oxide synthase and enhanced by nitric oxide donors. Mice with targeted deletion of the gene, for neuronal nitric oxide have urethral sphincters that do not relax in response to electrical stimulation. We hypothesise that augmentation of external sphincter nitric oxide could be an effective pharmacological treatment for DSD. Currently available nitric oxide donors such as glyceryl trinitrate or isosorbide mononitrate could be used to deliver nitric oxide to the urethral sphincter. The variable pharmacokinetics of these drugs combined with different modes of delivery (sublingual, buccal, or oral) could be used to achieve both short-term and long-term increases in concentrations of sphincter nitric oxide, thereby resulting in either acute or chronic lowering of urethral pressure. The safety and efficacy of this potential treatment for DSD needs to be established in clinical trials of men with spinal cord injures with DSD. PMID- 11425439 TI - Bioethical issues in caring for conjoined twins and their parents. PMID- 11425448 TI - Updates on the cytogenetics and molecular genetics of bone and soft tissue tumors. Mesothelioma. PMID- 11425449 TI - Problems with interphase fluorescence in situ hybridization in detecting BCR/ABL positive cells in some patients using a novel technique with extra signals. AB - Interphase fluorescence in situ hybridization (I-FISH) is frequently used to monitor the response to therapy in various hematological malignancies. We performed a comparison of I-FISH, metaphase FISH (C-FISH), conventional cytogenetics, spectral karyotyping (SKY) and PCR for the detection of the t(9;22) and the BCR/ABL rearrangement in 32 patients with chronic myelogenous leukemia (CML). FISH was done using a novel commercial probe set (VYSIS LSI BCR/ABL ES), which is designed to reduce the rate of false-positive results by marking the argininosuccinate synthetase (ASS) gene and thus providing an extra signal on chromosome 9. Our data indicate, that a substantial number of BCR/ABL-positive patients (n=5 patients, 3 with Ph, 2 with masked Ph) present negative results using this probe set in I-FISH analyses, because they did not fulfill the scoring criteria. In fact, the ASS region, which usually remains on 9q in Ph+ CML, appears to be lost or translocated. Due to these results we recommend that the initial diagnosis as well as the follow-up of patients with Ph+ leukemias should not be based on a single technique but should integrate results of cytogenetics and molecular biology. PMID- 11425450 TI - Analysis of genetic changes in rat endometrial carcinomas by means of comparative genomic hybridization. AB - Animals of the BDII inbred rat strain are known to be genetically predisposed to endometrial adenocarcinoma (EAC). Using them as models of human EACs, we studied tumors arising in F1 and F2 progeny from BDII animals crossed to animals from two other inbred strains, in which EACs were quite rare. In order to identify chromosomal regions exhibiting DNA copy number changes, comparative genomic hybridization (CGH) was applied in a series corresponding to 27 different solid tumors, most of which were classified as EACs, from these animals. The main findings from the study were that, although many different chromosomes were involved in copy number variation, some of the changes detected were recurrent and quite specific. Among specific changes found were gains in rat chromosome (RNO) regions 4q12 approximately q22, 6q14 approximately q16, and whole chromosome arms in some of the small metacentric chromosomes (e.g., RNO14, 16, and 18). RNO10 was involved in gain in the terminal and proximal regions. Each of these regions contains previously identified cancer-related genes representing possible candidates to be involved in the development of EAC. Furthermore, it was observed that there were clear differences in the pattern of copy number changes between tumors occurring in the two different crosses, and also between solid tumors and cell cultures. Endometrial cancer is the most common human gynecological cancer, but not much is known about specific genetic changes influencing this disease. Two genetic alterations that have been reported from human endometrial cancer are amplification of the ERBB2 gene and mutations in the 12 codon of the KRAS gene. One case of Erbb2 amplification was found but there were no Kras mutations in the rat material studied. We conclude that molecular genetic analysis of the rat BDII model will provide important new information about EAC in mammals. PMID- 11425451 TI - A highly aggressive primitive mesenchymal tumor with a translocation (1;19)(q12;q13.2). AB - Soft tissue sarcomas constitute a heterogeneous group of malignant tumors of mesenchymal origin, the classification of which may present a diagnostic challenge. We present here the cytological, histopathological, immunohistochemical, and cytogenetic findings of an unusual case of a highly aggressive sarcoma. Based on the morphology and the immunohistochemical profile, this primitive tumor and its metastases could not be conclusively classified as any of the defined subtypes of sarcomas, although the findings were suggestive of a variant of rhabdomyosarcoma. Cytogenetic characterization using G-banding, SKY, FISH, and CGH revealed almost identical chromosomal compositions of the primary tumor and the metastasis. The hypertetraploid karyotype was characterized by numerical imbalances as well as by an unbalanced translocation t(1;19)(q12;q13.2), which has not been previously reported. PMID- 11425452 TI - Philadelphia-positive chronic myelogenous leukemia with a 5q-- abnormality in a patient following interferon-alpha therapy. AB - In this report, we describe a rare 5q--/CML association in a patient with Ph positive chronic myelogenous leukemia (CML) who achieved complete cytogenetic response on interferon-alpha (IFN-alpha) treatment, but who developed a new clone in the blastic crisis. The patient was treated with interferon-alpha beginning in 1996 and a serial chromosome and molecular study was performed over the clinical course of the disease. The patient remained in complete hematologic and cytogenetic remission until November 1998, when a reverse transcriptase PCR study performed on the bone marrow and peripheral blood cells was negative for chimeric BCR/ABL mRNA. The treatment was discontinued until April 1999, when the patient developed acute transformation of the disease. In June 1999, cytogenetic examination showed the development of a new clone, consisting of the deletion of the long arm of chromosome 5 in addition to the standard Ph translocation. The unusual association of a Ph with an abnormality usually observed in a secondary myeloproliferative disease raises the question of whether the new finding is treatment-induced or part of the disease process and casually related to the acute transformation. PMID- 11425453 TI - Translocation (2;15)(p23;q21.2) and interstitial deletion of 7q in a case of low grade myxofibrosarcoma. AB - We describe a case of low-grade myxofibrosarcoma with the karyotype 46,XX,t(2;15)(p23;q21.2), del(7)(q?11.2q?22). Only six of these tumors have been previously studied and all were cytogenetically different. PMID- 11425454 TI - Spectral karyotyping study of chromosome abnormalities in human leukemia. AB - Chromosomal analysis plays an important role in the diagnosis, treatment and prognosis of human leukemia. Currently, the GTG-banding technique (G-banding) is the most commonly used diagnostic method in clinical cytogenetics. G-banding analysis of subtle chromosomal rearrangements or complex karyotypes with multiple markers can be inadequate because of poor chromosome morphology and/or an insufficient yield of analyzable metaphases. Fluorescence in situ hybridization (FISH) is a highly sensitive and specific method to detect chromosomal alterations. Conventional FISH is used optimally in instances where only one or a few abnormalities are investigated. Spectral karyotyping (SKY), a novel cytogenetic technique, has been developed to unambiguously display and identify all chromosomes at one time using a spectrum of 24 different colors. This report presents the use of SKY for examination of the entire karyotype in specimens with complex chromosomal abnormalities from three leukemia patients. Conventional cytogenetic analysis (G-banding) showed complex hyperdiploid clones with multiple markers in each case. SKY was able to clarify and identify additional cryptic chromosomal translocations [e.g., t(2;10), t(3;10), t(5;7), t(7;18), t(9;17), t(10;12), t(13;16)] insertions [e.g., ins(17;9), ins(20;Y)], duplications [e.g., i(8)(q10), dup(4)(q31q35)] and marker chromosomes in each case. This study demonstrates that the combination of SKY and G-band techniques results in a more complete characterization of the complex chromosomal aberrations seen in leukemia. PMID- 11425455 TI - Extensive cytogenetic heterogeneity in a benign retroperitoneal schwannoma. AB - A benign retroperitoneal schwannoma from a patient without prior exposure to radiotherapy or chemotherapy was analyzed by chromosome banding after short-term culture. An extensive intratumor heterogeneity in the form of 29 karyotypically related as well as unrelated clones was found. The aberrant clones were diploid or near-diploid and displayed both numerical and structural changes. All chromosomes, except 11, 16, and 20, were affected. Numerical changes included trisomies X, 7, 9, 17, and 18, and monosomies 13 and 18. No clonal loss of chromosome 22, the most characteristic abnormality in schwannomas of other locations, was, however, detected. The structural aberrations resulted in a total of 58 chromosomal breakpoints, with chromosomes 18, 1, and 15 participating in rearrangements most frequently, followed by chromosomes 14, 2, and 22. A striking finding was the clonal involvement of 18p11 in eight rearrangements affecting different chromosomes, suggesting alteration of telomeric function. The molecular mechanisms underlying the observed massive polyclonality in the schwannoma, particularly the presence of cytogenetically unrelated clones, are unknown and probably heterogeneous. PMID- 11425456 TI - Low frequency of genetic lesions in Wilms tumors by representational difference analysis. AB - Genomic representational difference analysis (RDA) was carried out on a total of nine Wilms tumors and one cystic partially differentiated nephroblastoma (CPDN; a sub-type of Wilms) to look for novel genetic deletions involving tumor suppressor genes. Genomic DNA from either short-term cultured Wilms tumor cells or a WT xenograft was used to create driver representations, and genomic DNA from matched normal kidney or normal kidney cells grown in short-term culture was used to create the tester. Genuine difference products were obtained from only one of the tumors. However, none of these fragments were found to be deleted in the original tumor biopsy, microdissected tumor or in the lung metastasis from this patient. It is, therefore, likely that the deletions were due to random losses associated with the genetic instability of the cultured cells from this particular tumor. We did not isolate difference products from any of the other tumors, showing that they did not have chromosomal losses, homozygous deletions or regions of LOH that were detectable by RDA. PMID- 11425457 TI - Genetic abnormalities specifically associated with varying metastatic potential of prostate cancer cell lines as detected by comparative genomic hybridization. AB - Established recently are two in vivo prostate tumor progression models in which subclones of the PC3M and LNCaP cell lines were selected for varying growth characteristics and metastatic potential after successive orthotopic implantation in the prostate of nude mice. In this study, we used comparative genomic hybridization (CGH) to compare the chromosomal abnormalities between the parental cell lines and their respective variants and to determine if specific chromosomal abnormalities can be identified that are associated with different growth properties. PC3M and its derivative cell lines PC3M-Pro4 and PC3M-LN4 shared gains of 8q22--qter, 10q21--q22, and Xq27--qter and loss of 13q33--qter. PC3M Pro4, a derivative line that produced significantly larger tumors in the prostate, had a unique gain of 3q13. In contrast, PC3M-LN4, the derivative line that produced significantly larger metastatic tumors in the lymph nodes and had higher incidences of distant metastases, had a specific gain of 1q21--q22 and losses of 10q23--qter and 18q12--q21. In the second in vivo model, LNCaP and its derivative cell lines shared gain of 3q27--qter and loss on 13q21--qter. The derivative line that produced significantly larger tumors in the prostate, LNCaP Pro5, had a unique gain on 13q12--q13. In comparison, LNCaP-LN3, a derivative line that had a significantly higher incidence of lymph node metastases and produced significantly larger metastatic tumors in the lymph nodes, had specific losses of 16q23--qter and 21q. Interestingly, some regions of loss (e.g., 10q23- >qter, 16q23-->qter, and 18q12-->q21) detected in the variant cell lines correlated well with abnormalities seen in clinical prostate cancer cases. Thus, our data suggest not only that these cell lines are relevant in vivo models for prostate cancer progression, but also that CGH is a valuable tool for uncovering chromosomal regions that are important for aggressive growth and metastasis of prostate cancer cells. PMID- 11425458 TI - Conformational differences in the 3-D nanostructure of the immunoglobulin heavy chain locus, a hotspot of chromosomal translocations in B lymphocytes. AB - Spectral precision distance microscopy was utilized to detect small but nonetheless consistently present conformational differences between the immunoglobulin heavy-chain gene clusters (IgH) that reside on the two chromosome 12 homologues in all diploid cells of the mouse. The euclidian distance (i.e., the mean arithmetic three-dimensional [3-D] distance) between the 5' most IgH gene, C mu, and the 3' most IgH gene, C alpha, was used as the indicator to define the co-presence of a condensed IgH domain and a relaxed IgH domain in the same cell. In normal and malignant B cells in which IgH is actively rearranged and transcribed, the C mu/C alpha distance (genomic distance approximately 180 kb) was found to range from 87.5 to 121 nm on the condensed IgH domain and from 154 to 207 nm on the relaxed IgH domain. In non-B cells (fibroblasts, neutrophils, and macrophages), in which IgH is inactive, the C mu/C alpha distance was found to range from 136 to 154 nm on the condensed IgH domain and from 250 to 292 nm on the related IgH domain. These results suggested that conformational differences that may predispose the relaxed IgH domain for illegitimate genetic recombinations, such as chromosomal translocations, are likely to exist in many cell types, including B cells. However, in B lymphocytes this structural predisposition may conspire with the lineage-specific ability to activate proto-oncogenes (after juxtaposition to IgH) to positively affect the preferential involvement of the relaxed IgH domain in chromosomal translocations. Additional studies are warranted to validate this working hypothesis. PMID- 11425459 TI - Amplification of ERBB2, RARA, and TOP2A genes in a myelodysplastic syndrome transforming to acute myeloid leukemia. AB - A patient is described with myelodysplastic syndrome (MDS) progressing to acute myeloid leukemia (AML) FAB M4. Cytogenetic analysis revealed an unusual rearrangement between chromosomes 9 and 17, leading to a dicentric chromosome with an insertion of material of unknown origin between both chromosomes. By fluorescence in situ hybridization (FISH), the insertion was shown to be an amplification of part of 17q, involving ERBB2, RARA, and TOP2A genes. The median copy number of ERBB2, RARA, and TOP2A genes in the tumor cells was six (range: 4- 10). Only one copy of the MPO gene at 17q21.3 was detected, suggesting a deletion of the telomeric part of 17q. To our knowledge, this is the first report of a 17q amplification in AML. PMID- 11425460 TI - Complex variant t(4;11) characterized by fluorescence in situ hybridization in infant acute lymphoblastic leukemia. AB - A 6-month-old girl was diagnosed with acute lymphoblastic leukemia (ALL). Chromosome analysis of bone marrow aspirate showed 46,XX,t(4;11)(q21;q23) with an atypical appearance of the 11p on the der(11) chromosome. FISH studies to fully characterize the translocation utilised 8 probes: whole chromosome painting probes for chromosome 11 and chromosome 4; separate chromosome 11 short arm and long arm paints; specific subtelomere probes from 11p, 11q, and 4q; MLL gene probe. Taken together, the results indicated a two-step abnormality: an initial standard t(4;11)(q21;q23), followed by another t(4;11)--this time, between the two derivative chromosomes. The MLL gene was split by the first translocation and its position altered by the second. PMID- 11425461 TI - A novel method for eliminating the melanin pigments from melanoma cells undergoing cytogenetic analysis in cases of uveal melanoma. AB - A method has been developed to eliminate melanin pigments from chromosomal plates of uveal melanoma tumors. On preparing the chromosomal plates of tumor masses of posterior uveal melanoma for cytogenetic analysis in intermediate pigmented population, it was found that the melanin pigments obscured the cells and thereby the chromosomes. Thus, cytogenetic analysis could not be carried out. Hence, a method was developed in our laboratory to eliminate these obscuring pigments. Several different attempts were made to eliminate the melanin pigments. Finally, washing the cells with phosphate buffered saline before the harvesting stage served the purpose. After harvesting and preparing the chromosomal plates, the cells were found to be devoid of melanin pigments. The protocol would help researchers trying to carry out cytogenetic analysis on melanoma tumor masses in populations with intermediate to dark pigmentation. PMID- 11425462 TI - BCL10 somatic mutations rarely occur in gastric lymphoma: detection of high frequency of polymorphisms in BCL10 coding region. AB - The BCL10 gene, recently isolated due to its involvement in the t(1;14)(p22;q32) of mucosa-associated lymphoid tissue B cell non-Hodgkin lymphoma (MALToma), was shown to have frequent somatic mutations and short deletions within the coding region in MALToma and a variety of other lymphomas and solid tumors. These observations have been recently questioned. In this study, we examined BCL10 gene mutations by direct sequencing of the entire coding region of the BCL10 gene, amplified from paired normal and tumor genomic DNAs, as well as tumor cDNAs, in 23 cases of primary gastric B cell non-Hodgkin lymphomas, comprising of 6 cases of MALToma and 17 cases of diffuse large cell (DLC) lymphoma. Heterozygosity due to three types of known polymorphisms in codon 5 (17.3%), codon 8 (21.7%), and codon 213 (8.6%) were observed in both normal germline DNA and tumor DNAs and tumor cDNAs in individual cases. In one case (4.3%) G/C heterozygosity in codon 8 in normal germline DNA was reduced to homozygosity (LOH) in tumor DNA and cDNA. Mutations inactivating BCL10 gene product function were not found in any of these cases. Moreover, post-transcriptional alterations were not indicated by abnormalities in BCL10 mRNA sequence in tumor cDNAs in these gastric lymphoma cases. Our results show that somatic mutations in the BCL10 gene rarely occur in gastric lymphoma and indicate that this gene is unlikely to be of pathogenetic significance in the majority of gastric lymphomas. PMID- 11425463 TI - Viral vectors as potential HIV-1 vaccines. AB - Vaccine vectors based on recombinant viruses have great promise to play an important role in the development of an effective HIV-1 vaccine. Within the last 10 years a wide range of viruses have been investigated for their ability to express protein(s) from foreign pathogens and to induce specific immunological responses against these antigen(s) in vivo. Each viral vector has its own unique biological characteristics and thus far none of them has proven to be an ideal candidate as a vaccine vehicle for HIV-1. This review focuses on both replication competent and non-replication competent viral vectors as a potential HIV-1 vaccine. Other approaches for the development of an HIV-1 vaccine are reviewed elsewhere and are beyond the scope of this review. PMID- 11425464 TI - Expression profiles of low molecular mass heat shock proteins by Mycobacterium avium and Mycobacterium intracellulare. AB - Closely related non-tuberculous mycobacterial species, Mycobacterium avium and Mycobacterium intracellulare, were compared for the profiles of their production of low molecular mass heat shock proteins at 45 degrees C, by performing polyacrylamide gel electrophoresis analysis of bacterial cell lysate proteins. All of the M. intracellulare but not M. avium strains potently increased the production of the 18-kDa heat shock protein, when cultured at 45 degrees C. Half of the M. intracellulare strains with high sensitivity to 45 degrees C produced not only the 18-kDa heat shock protein but also the 16-kDa heat shock protein at 45 degrees C. These findings indicate that M. avium and M. intracellulare differentially respond to 45 degrees C heat shock in terms of the production of low molecular mass heat shock proteins. PMID- 11425465 TI - Comparative genomics for identification of clone-specific sequence blocks in Streptococcus pneumoniae. AB - The partial genome sequences of a serotype 3 and a serotype 2 pneumococcal strain were compared to the complete type 4 pneumococcal genome. Over 500000 and 150000 base pairs of the partial genome data, obtained from published patents, were analysed respectively. Global alignment showed that nearly the whole genome is highly conserved in accordance with data of multilocus sequence typing of housekeeping genes. The search for clone-specific genes revealed 17 new open reading frames in the type 3 strain, while no new open reading frame was detected in the type 2 strain. Allelic variation of genes was restricted by the use of crude sequence data, but still permitted identification of some new alleles and the observation that all surface proteins present in the partial genome data were highly conserved. In both strains we observed also a variety of chromosomal rearrangements and variations due to mobile genetic elements. All together, this comparative genomic approach gives a genome-based overview of strain relatedness and a prospective on what could be expected when sequencing other pneumococcal strains. PMID- 11425466 TI - Inhibition of epithelial cell apoptosis by Porphyromonas gingivalis. AB - Porphyromonas gingivalis is periodontal pathogen that is capable of invading gingival epithelial cells (GECs). Apoptotic responses of primary cultures of GECs to P. gingivalis were investigated with a DNA fragmentation ELISA assay. P. gingivalis induced a transient increase in GEC DNA fragmentation; however, after prolonged incubation GECs did not undergo apoptosis. Furthermore, P. gingivalis blocked apoptosis in GECs following stimulation with camptothecin. Immunoblotting of GECs with Bcl-2 or Bax antibodies showed that P. gingivalis up-regulated Bcl-2 levels in GECs, whereas Bax levels were transiently elevated and declined after 24 h stimulation. Streptococcus gordonii did not affect levels of either molecule. RT-PCR demonstrated that induction of Bcl-2 occurs at the transcriptional level. The results suggest that P. gingivalis can inhibit apoptosis in GECs by up-regulation of the anti-apoptotic molecule Bcl-2. The prevention of host cell apoptosis may represent a strategy for P. gingivalis survival within invaded GECs. PMID- 11425467 TI - Identification of a cadmium-induced gene in Mycobacterium bovis and Mycobacterium tuberculosis. AB - A 17-kDa protein (CadI) was induced by cadmium in Mycobacterium bovis and Mycobacterium tuberculosis. Comparison of the N-terminal sequence from M. bovis CadI with the annotated M. tuberculosis genome database identified Rv2641 as the encoding gene. Long and short promoter fragments from M. bovis cadI were fused to the lacZ reporter gene in pYUB76. Only the long fragment directed cadmium inducible activity when electroporated into M. bovis. The cadI promoter has potential for both constitutive and inducible expression studies in M. bovis and M. tuberculosis. PMID- 11425468 TI - Restriction fragment length dimorphism-PCR method for the detection of extended spectrum beta-lactamases unrelated to TEM- and SHV-types. AB - The diagnostic ability of the restriction fragment length dimorphism-polymerase chain reaction (RFLD-PCR) method was evaluated. Seven primer pairs, newly designed from 44 beta-lactamase genes encoding extended-spectrum beta-lactamases not related to TEM- and SHV-types, were used to differentiate OXA-2, FOX-3, CMY 3, IMP-1, and IMI-1 beta-lactamases. The RFLD-PCR was carried out successfully, and these genes were differentiated by the sizes of their PCR products and by the difference in restriction fragment length when each amplicon was digested with a unique restriction enzyme. This discriminatory detection of the genes was confirmed by sequencing the PCR products. PMID- 11425469 TI - Transformation of an oral bacterium via chromosomal integration of free DNA in the presence of human saliva. AB - Transformation of Streptococcus gordonii DL1 by free DNA was studied in human saliva. Competent S. gordonii could be transformed in vitro with plasmid DNA that had been taken into the human mouth. Transformation also occurred with a plasmid that cannot replicate in S. gordonii, but that has a region of chromosomal homology, by integration into the bacterial chromosome, although linearised plasmid DNA gave no transformants. Linear chromosomal DNA fragments did however transform S. gordonii/Tn916 efficiently in saliva when regions of homology with the recipient chromosome flanked the marker gene. These findings are discussed in relation to the potential for acquisition of DNA sequences, including genetically modified DNA, by gut and oral bacteria. PMID- 11425470 TI - Antifungal activity of a novel endochitinase gene (chit36) from Trichoderma harzianum Rifai TM. AB - A novel 36-kDa endochitinase named chit36 has been isolated and characterized from Trichoderma harzianum Rifai TM. Partial amino acid sequences from the purified protein were used to clone the fungal cDNA, based on polymerase chain reaction with degenerate primers. The complete open reading frame encodes a 344 amino acid protein which shows 84% similarity to a putative chitinase from Streptomyces coelicolor. Chit36 was overexpressed under the pki1 constitutive promoter from Trichoderma reesei via biolistic transformation of T. harzianum TM. Stable transformants showed expression and endochitinase activity of chit36 in glucose-rich medium. Culture filtrates containing secreted CHIT36 as the sole chitinolytic enzyme completely inhibited the germination of Botrytis cinerea conidia. Growth of Fusarium oxysporum f. sp. melonis and Sclerotium rolfsii were significantly inhibited on agar plates on which the Trichoderma transformants had previously been grown. PMID- 11425471 TI - Lentinula edodes produces a multicomponent protein complex containing manganese (II)-dependent peroxidase, laccase and beta-glucosidase. AB - A multicomponent protein complex containing manganese (II)-dependent peroxidase, laccase and beta-glucosidase was isolated from culture extracts of the white rot basidiomycete Lentinula edodes. This protein complex showed a single protein band on native polyacrylamide gel electrophoresis (PAGE). On sodium dodecyl sulfate (SDS)-PAGE, however, it displayed three major bands and several additional minor bands ranging in size from 60 kDa to 180 kDa, suggesting it being a complex of six to eight different proteins. The molecular mass of this complex was estimated to be approximately 660 kDa from the elution position of gel filtration. This enzyme complex was effective in transforming environmentally persistent xenobiotics, pentachlorophenol and 2,5-dichlorophenol. PMID- 11425472 TI - Molecular cloning and identification of UTR1 of a yeast Saccharomyces cerevisiae as a gene encoding an NAD kinase. AB - UTR1 of the yeast Saccharomyces cerevisiae was cloned from the genomic DNA by polymerase chain reaction and expressed in Escherichia coli. Characterization of the purified UTR1p revealed that UTR1p is a NAD kinase consisting of six identical subunits with a molecular mass of 60 kDa. UTR1p specifically phosphorylated NAD in the presence of ATP, dATP, or CTP as phosphoryl donors, and was most active at pH 8.0, 30 degrees C. Km values of UTR1p for NAD and ATP were determined to be 0.50 mM and 0.60 mM, respectively. PMID- 11425473 TI - Intracellular pH-dependent efflux of the fluorescent probe pyranine in the yeast Yarrowia lipolytica. AB - 8-Hydroxypyrene-1,3,6-trisulfonic acid (pyranine) can be used as a vital intracellular pH (pH(i)) indicator. In the yeast Yarrowia lipolytica, a partial efflux of the probe was detected by using the pH-independent wavelength of 415 nm. A simplified correction of the fluorescent signals was applied, enabling to show for this species a good near-neutral pH(i) maintenance capacity in a pH 3.9 medium. Octanoic acid, which is known to have toxic effects on yeast, decreased the pH(i) and increased the 260-nm-absorbing compounds leakage. However, this acid inhibited the fluorescent probe efflux linearly with its concentration suggesting a pH(i)-dependent efflux of pyranine from cells. PMID- 11425474 TI - Restriction enzyme BstZ17I is sensitive to cytosine methylation. AB - The cleavage patterns of a subset of restriction enzymes are blocked or impaired when a methylated CpG is overlapped with either the 5' or 3' end of the canonical restriction site. BstZ17I restriction endonuclease is a blunt-end cutter, which recognises the hexanucleotide sequence GTA(downward arrow)TAC. In this report, I show that the BstZ17I restriction enzyme is sensitive to cytosine methylation. Using both in vitro-methylated episomal plasmids and lambdaDNA, I demonstrate that the BstZ17I restriction enzyme is sensitive to cytosine methylation that occurs 3' and/or 5' of the canonical recognition sequence. PMID- 11425475 TI - Biosynthesis of fatty acids and triacylglycerols by 2,6,10,14-tetramethyl pentadecane-grown cells of Nocardia globerula 432. AB - Nocardia globerula strain 432 was able to synthesize triacylglycerols (TAG) during cultivation on 2,6,10,14-tetramethyl pentadecane (pristane) under nitrogen limiting conditions. Within these cells, 4,8,12-trimethyl tridecanoic acid was the major fatty acid detected. Fatty acids with an odd number of carbon atoms and minor amounts of even-numbered fatty acids were also observed. Experiments carried out with acrylic acid, an inhibitor of beta-oxidation, suggested that odd numbered fatty acids such as C15:0, C17:0 and 10-methyl C17:0 were synthesized de novo using propionyl-CoA, the beta-oxidation product, as precursor. Although N. globerula 432 incorporated mainly straight chain fatty acids into TAG, the branched fatty acid 4,8,12-trimethyl tridecanoic acid also appeared, to some extent, in the acylglycerols. The importance of TAG biosynthesis by pristane grown cells of N. globerula strain 432 is discussed. PMID- 11425476 TI - Role of the ftsA1p promoter in the resistance of mucoid mutants of Salmonella enterica to mecillinam: characterization of a new type of mucoid mutant. AB - Mucoid mutants of Salmonella enterica serovar Typhimurium isolated by resistance to mecillinam include lon (27%) and rcsC (8%) mutants but the most frequent class (65%) is affected in a new gene (mucM) located at centisome 76. mucM cells are shorter than mucM+ cells and rcsB mutations normalize size and response to mecillinam. Expression of ftsA1p, the ftsA-ftsZ promoter submitted to RcsB stimulation, is greatly increased in mucM mutants, and this expression is dependent on RcsB and ftsA1p. It is proposed that the mucM product interferes with RcsB activation. Mucoidy results from the activation of cps genes and mecillinam resistance from ftsA-ftsZ overexpression, both traits caused by the increased activity of the RcsB effector. The same mechanism seems to be responsible for the resistance of mucoid rcsC mutants to mecillinam but the resistance of lon mutants is not dependent on RcsB and so responds to a different cause. PMID- 11425477 TI - Rhodobacter capsulatus nifA mutants mediating nif gene expression in the presence of ammonium. AB - Expression of nitrogen fixation genes in Rhodobacter capsulatus is repressed by ammonium at different regulatory levels including an NtrC-independent mechanism controlling NifA activity. In contrast to R. capsulatus NifA, heterologous NifA proteins of Klebsiella pneumoniae and Rhizobium meliloti, respectively, were not subjected to this posttranslational ammonium control in R. capsulatus. The characterization of ammonium-tolerant R. capsulatus NifA1 mutants indicated that the N-terminal domain of NifA was involved in posttranslational regulation. Analysis of a double mutant carrying amino acid substitutions in both the N terminal domain and the C-terminal DNA-binding domain gave rise to the hypothesis that an interaction between these two domains might be involved in ammonium regulation of NifA activity. Western analysis demonstrated that both constitutively expressed wild-type and ammonium-tolerant NifA1 proteins exhibited high stability and accumulated to comparable levels in cells grown in the presence of ammonium excluding the possibility that proteolytic degradation was responsible for ammonium-dependent inactivation of NifA. PMID- 11425478 TI - Plate-based dormancy culture system for Mycobacterium smegmatis and isolation of metronidazole-resistant mutants. AB - Mycobacterium smegmatis is an obligate aerobe. However, growth analyses in oxygen limited liquid cultures have shown that the bacillus is able to survive anoxia with a half-life of 4 days by shifting down to a drug-resistant, dormant state. Metronidazole is the first lead against dormant bacilli and shows selective toxicity for this physiological state. Here, we report a plate-based dormancy culture system employing anoxic jars for M. smegmatis. Its usefulness for the genetic analysis of dormancy was demonstrated by isolating the first metronidazole-resistant mutants. Highly resistant mutants formed slightly yellow (as opposed to creamy) colonies. Furthermore, high-level metronidazole resistance correlated with an increased half-life of 12 days under anoxic conditions. This suggests a link between metronidazole susceptibility and anaerobic survival. PMID- 11425479 TI - Heterologous overexpression of glucose dehydrogenase from the halophilic archaeon Haloferax mediterranei, an enzyme of the medium chain dehydrogenase/reductase family. AB - The first gene encoding a glucose dehydrogenase (GDH) from a halophilic organism has been sequenced. Amino acid sequence alignments of GDH from Haloferax mediterranei show a high degree of homology with the thermoacidophilic GDHs and with other enzymes from the medium chain dehydrogenase/reductase family. Heterologous overexpression using the mesophilic organism Escherichia coli as the host has been performed and the expression product was obtained as inclusion bodies. To obtain the halophilic enzyme in its native form refolding and reactivation in a saline environment were required. A pure and highly concentrated sample of the enzyme was obtained using a purification procedure based on the protein's halophilicity. This method may be useful as a general procedure for purifying other halophilic proteins from mesophilic hosts. PMID- 11425480 TI - Lipid modification of prelipoproteins is dispensable for growth in vitro but essential for virulence in Streptococcus pneumoniae. AB - A Deltalgt (Lgt, lipoprotein diacylglyceryl transferase) isogenic mutant was obtained which indicates that lgt is not essential for cell growth in vitro, like in the Gram-positive bacterium Bacillus subtilis, but unlike in the proteobacteria Escherichia coli and Salmonella typhimurium. The mutation was transduced to a virulent strain. A 5 log attenuation was observed in a respiratory tract model of infection. Metabolic labeling by [U-14C]palmitate revealed the presence of eight to ten lipoproteins in the wild-type strain only, with molecular masses between 15 and 80 kDa. Our findings suggest a major difference in the role of lipoproteins in Gram-positive bacteria versus the proteobacteria. PMID- 11425481 TI - Plasmid-encoded catalase KatA, the main catalase of Pseudomonas fluorescens strain Cb36. AB - The plasmid-state catalase gene katA of the phenol gradative Pseudomonas fluorescens isolate Cb36 has been characterized and shown to be the major catalase of this strain. The predicted amino acid sequence of KatA revealed significant similarity with the catalase sequence from Neisseria meningitidis and has probably the non-pseudomonad origin. The specific activity of catalase was investigated and elevated catalase activity was found in stationary phase cells. The consensus sequence for promoters recognized by the stationary phase sigma factor sigma(s) was found 212 bp upstream of the putative ATG start codon. The ability of KatA to detoxify a high concentration of hydrogen peroxide and protect Pseudomonas putida and Escherichia coli cells was shown. PMID- 11425482 TI - Identification, classification and phylogeny of the Aspergillus section Nigri inferred from mitochondrial cytochrome b gene. AB - The partial mitochondrial cytochrome b gene from 32 strains of 12 species belonging to Aspergillus section Nigri was amplified by the polymerase chain reaction and sequenced directly. Using 402 nucleotide characters, nucleotide based and amino acid-based phylogenetic trees were inferred and the genetic divergence among the species was evaluated. Based on analyses of the 402-bp nucleotide and 133-amino acid sequences, strains were divided into 11 DNA types and five amino acid types. Aspergillus niger and Aspergillus awamori showed different amino acid sequences. A. niger clade included A. niger var. niger and Aspergillus ficuum. A. awamori clade included A. awamori, Aspergillus phoenicis, Aspergillus pulverulentus, Aspergillus tubingensis, Aspergillus foetidus, and two varieties of A. niger, var. nanus and var. intermedius. Two varieties of A. niger will be reclassified. One strain of A. phoenicis and one strain of Aspergillus carbonarius were reidentified. PMID- 11425483 TI - Protein synthesis patterns in Acinetobacter calcoaceticus induced by phenol and catechol show specificities of responses to chemostress. AB - The proteins induced in Acinetobacter calcoaceticus by the potentially toxic growth substrates phenol and catechol were analyzed by 2D-electrophoresis of cell extracts and compared with those induced by heat shock and oxidative stress. Although both aromatic compounds are quite similar, the only difference being that catechol has an additional hydroxyl group, the responses obtained differed considerably. Phenol has greater lipophilicity and mainly induced heat shock proteins, whereas catechol, which causes the production of reactive oxygen species, predominantly induced oxidative stress proteins. Furthermore, some special proteins were induced by phenol or catechol, which might be useful as biomarkers for chemostress, and could be involved in the catalytic degradation of potentially toxic compounds. PMID- 11425484 TI - UVA irradiation induces L-isoaspartyl formation in melanoma cell proteins. AB - It has been reported that UVA effects are partly mediated by production of reactive oxygen species. Moreover, oxidative stress increases protein damage, involving the occurrence of isoaspartyl residues, a product of protein deamidation/isomerization reactions. This work was undertaken in order to study the effects of UVA irradiation, mediated by oxidation, on sensitive protein targets. Melanoma cells exposed to UVA rays have been chosen as a model for monitoring the occurrence of L-isoaspartyl sites. A dramatic increase of these abnormal residues, specifically recognized and methylated by the enzyme L isoaspartate(D-aspartate) O-methyltransferase (PCMT; EC 2.1.1.77), can be detected after exposure of M14 cells to raising doses of UVA. The effect of UVA on NO and TBARS accumulation, as well as on DNA fragmentation, has also been investigated. NO formation parallels the increase in isoaspartyl formation, while lipid peroxidation occurs only at the highest UVA doses. No DNA fragmentation has been detected under the employed experimental conditions. These results, as a whole, indicate that protein damages are one of the early events on UVA-induced cell injury. The endogenous activity of PCMT remains remarkably stable under UVA treatment, suggesting that this enzyme might play a crucial role in the repair and/or disposal of damaged proteins in UVA-irradiated cells. PMID- 11425485 TI - Attenuation of radiation-induced genomic instability by free radical scavengers and cellular proliferation. AB - To investigate the mechanisms of radiation-induced chromosomal instability, cells were irradiated in the presence of the free radical scavengers DMSO, glycerol, or cysteamine, in the presence of DMSO while frozen, or held in confluence arrest post-irradiation to permit cells to repair potentially lethal DNA damage. Clones derived from single progenitor cells surviving each treatment were then analyzed for the subsequent development of chromosomal instability. The presence of scavengers (+/- freezing) during irradiation, and the recovery from potentially lethal damage after irradiation led to an increase in cell survival that was accompanied by a decrease in the initial yield of chromosomal rearrangements. Furthermore, analysis of over 400 clones and 80,000 metaphases indicates that these same treatments reduced the incidence of instability at equitoxic doses when compared to controls irradiated in the absence of scavengers at ambient temperature. Results suggest that preventing reactive species from damaging DNA, promoting chemical repair of ionized DNA intermediates, or allowing enzymatic removal of genetic lesions, represent measures that reduce the total burden of DNA damage and reduce the subsequent onset of radiation-induced genomic instability. PMID- 11425486 TI - Glyoxal and methylglyoxal trigger distinct signals for map family kinases and caspase activation in human endothelial cells. AB - Carbonyl compounds with diverse carbon skeletons may be differentially related to the pathogenesis of vascular diseases. In this study, we compared intracellular signals delivered into cultured human umbilical vein endothelial cells (HUVECs) by glyoxal (GO) and methylglyoxal (MGO), which differ only by a methyl group. Depending on their concentrations, GO and MGO promoted phosphorylations of ERK1 and ERK2, which were blocked by the protein-tyrosine kinase (PTK) inhibitors herbimycin A and staurosporine, thereby being PTK-dependent. GO and MGO also induced phosphorylations of JNK, p38 MAPK, and c-Jun, either PTK-dependently (GO) or -independently (MGO). Next, we found that MGO, but not GO, induced degradation of poly(ADP-ribose) polymerase (PARP) as the intracellular substrate of caspase 3. Curcumin and SB203580, which inhibit JNK and p38 MAPK signaling pathways, but not herbimycin A/staurosporine, prevented the MGO-induced PARP degradation. We then found that MGO, but not GO, reduced the intracellular glutathione level, and that cysteine, but not cystine, inhibited the MGO-mediated activation of ERK, JNK, p38 MAPK, or c-Jun more extensively than did lysine or arginine. In addition, all the signals triggered by GO and MGO were blocked by amino guanidine (AG), which traps carbonyls. These results demonstrated that GO and MGO triggered two distinct signal cascades, one for PTK-dependent control of ERK and another for PTK-independent redox-linked activation of JNK/p38 MAPK and caspases in HUVECs, depending on the structure of the carbon skeleton of the chemicals. PMID- 11425487 TI - New synthetic flavonoids as potent protectors against doxorubicin-induced cardiotoxicity. AB - A series of 3,7-disubstituted-2(3',4'-dihydroxyphenyl) flavones has been studied as potential cardioprotective agents in doxorubicin antitumor therapy. The influence of substituents on the 3 and 7 position of the flavone nucleus on antioxidant activity cytotoxicity and cardioprotective properties was explored to improve the activity of our lead compound 7-monohydroxyethylrutoside. In the protection against Fe(2+)/vitamin C-induced microsomal lipid peroxidation (LPO assay), IC(50) values ranged from 0.2 to 37 microM. In general, the 3-substituted flavones were the most potent compounds in this assay. The cytotoxicity of the new compounds was tested (up to 250 microM) in hepatocytes. LDH leakage ranged from 2.6-29.2%, whereas the GSH concentrations decreased to 87.3-41.3%. Only four compounds out of this series protected the isolated mouse left atrium against doxorubicin-induced toxicity. Because of the positive inotropic effect of 8d (N (3-(3',4'-dihydroxyflavon-7-yl)oxypropyl)-N,N,N-trimethylammonium chloride) and 10c (3-hydroxyethoxy-7,3',4'-trihydroxyflavone) on the atrium, compounds 9i (3',4'-dihydroxy-3-glucosylflavone) and 10d (N-(3-(7,3',4'-trihydroxyflavon-3 yl)oxypropyl)-N,N,N-trimethylammonium chloride) were selected to be evaluated as cardioprotective agents in vivo. PMID- 11425488 TI - Upregulation of oxidant-induced VEGF expression in cultured keratinocytes by a grape seed proanthocyanidin extract. AB - Angiogenesis plays a central role in wound healing. Among many known growth factors, vascular endothelial growth factor (VEGF) is believed to be the most prevalent, efficacious, and long-term signal that is known to stimulate angiogenesis in wounds. The wound site is rich in oxidants such as hydrogen peroxide mostly contributed by neutrophils and macrophages. Proanthocyanidins or condensed tannins are a group of biologically active polyphenolic bioflavonoids that are synthesized by many plants. This study provides first evidence showing that natural extracts such as grape seed proanthocyanidin extract containing 5000 ppm resveratrol (GSPE) facilitates oxidant-induced VEGF expression in keratinocytes. Using a ribonuclease protection assay (RPA), the ability of GSPE to regulate oxidant-induced changes in several angiogenesis-related genes were studied. While mRNA responses were studied using RPA, VEGF protein release from cells to the culture medium was studied using ELISA. Pretreatment of HaCaT keratinocytes with GSPE upregulated both hydrogen peroxide as well as TNF-alpha induced VEGF expression and release. The current results suggest that GSPE may have beneficial therapeutic effects in promoting dermal wound healing and other related skin disorders. PMID- 11425489 TI - Carotenoids as antioxidants: spin trapping EPR and optical study. AB - The role of several natural and synthetic carotenoids as scavengers of free radicals was studied in homogeneous solutions. A set of free radicals: *OH, *OOH, and *CH(3) were generated by using the Fenton reaction in dimethyl sulfoxide. It was shown that the spin trapping technique is more informative than optical methods for the experimental conditions under study. 5,5-Dimethyl-pyrroline-N oxide (DMPO) and N-tert-butyl-alpha-phenylnitrone (PBN) were used as spin traps for the EPR studies. The results show that the scavenging ability of the carotenoids towards radical *OOH correlates with their redox properties. PMID- 11425490 TI - Beneficial effects of alpha-lipoic acid and ascorbic acid on endothelium dependent, nitric oxide-mediated vasodilation in diabetic patients: relation to parameters of oxidative stress. AB - The impairment of nitric oxide (NO)-mediated vasodilation in diabetes has been attributed to increased vascular oxidative stress. Lipoic acid has been shown to have substantial antioxidative properties. The aim of this study was to assess the effect of lipoic acid on NO-mediated vasodilation in diabetic patients in comparison with the well-recognized effect of ascorbic acid. Using venous occlusion plethysmography, we examined the effects of lipoic acid (0.2 mM) and ascorbic acid (1 and 10 mM) on forearm blood flow responses to acetylcholine, sodium nitroprusside and concomitant infusion of the NO-inhibitor, N(G) monomethyl-L-arginine, in 39 diabetic patients and 11 control subjects. Plasma levels of antioxidants and parameters of lipid peroxidation were measured and correlated to endothelial function tests. Lipoic acid improved NO-mediated vasodilation in diabetic patients, but not in controls. NO-mediated vasodilation was improved by ascorbic acid at 10 mM, but not 1 mM. Improvements of endothelial function by ascorbic acid and lipoic acid were closely related. The beneficial effects of lipoic acid were positively related to plasma levels of malondialdehyde and inversely related to levels of ubiquinol-10. These findings support the concept that oxidative stress contributes to endothelial dysfunction and suggest a therapeutic potential of lipoic acid particularly in patients with imbalance between increased oxidative stress and depleted antioxidant defense. PMID- 11425491 TI - Comparison of low-density lipoprotein modification by myeloperoxidase-derived hypochlorous and hypobromous acids. AB - Myeloperoxidase (MPO), a heme enzyme secreted by activated phagocytes, catalyzes the oxidation of halides to hypohalous acids. At plasma concentrations of halides, hypochlorous acid (HOCl) is the major strong oxidant produced. In contrast, the related enzyme eosinophil peroxidase preferentially generates hypobromous acid (HOBr). Since reagent and MPO-derived HOCl converts low-density lipoprotein (LDL) to a potentially atherogenic form, we investigated the effects of HOBr on LDL modification. Compared to HOCl, HOBr caused 2-3-fold greater oxidation of tryptophan and cysteine residues of the protein moiety (apoB) of LDL and 4-fold greater formation of fatty acid halohydrins from the lipids in LDL. In contrast, HOBr was 2-fold less reactive than HOCl with lysine residues and caused little formation of N-bromamines. Nevertheless, HOBr caused an equivalent increase in the relative electrophoretic mobility of LDL as HOCl, which was not reversed upon subsequent incubation with ascorbate, in contrast to the shift in mobility caused by HOCl. Similar apoB modifications were observed with HOBr generated by MPO/H(2)O(2)/Br(-). In the presence of equivalent concentrations of Cl(-) and Br(-), modifications of LDL by MPO resembled those seen in the presence of Br(-) alone. Interestingly, even at physiological concentrations of the two halides (100 mM Cl(-), 100 microM Br(-)), MPO utilized a portion of the Br(-) to oxidize apoB cysteine residues. MPO also utilized the pseudohalide thiocyanate to oxidize apoB cysteine residues. Our data show that even though HOBr has different reactivities than HOCl with apoB, it is able to alter the charge of LDL, converting it into a potentially atherogenic particle. PMID- 11425492 TI - Expression of glutathione-S-transferase isozyme in the SY5Y neuroblastoma cell line increases resistance to oxidative stress. AB - Glutathione-S-transferases (GSTs) are a superfamily of enzymes that function to catalyze the nucleophilic attack of glutathione on electrophilic groups of a second substrate. GSTs are present in many organs and have been implicated in the detoxification of endogenous alpha, beta unsaturated aldehydes, including 4 hydroxynonenal (HNE). Exogenous GST protects hippocampal neurons against HNE in culture. To test the hypothesis that overexpression of GST in cells would increase resistance to exogenous or endogenous HNE induced by oxidative stress, stable transfectants of SY5Y neuroblastoma cells with GST were established. Stable GST transfectants demonstrated enzyme activities 13.7 times (Clone 1) and 30 times (Clone 2) higher than cells transfected with vector alone. GST transfectants (both Clones 1 and 2) demonstrated significantly (p <.05) increased resistance to ferrous sulfate/hydrogen peroxide (20.9% for Clone 1; 46.5% for Clone 2), amyloid beta-peptide (12.2% for Clone 1; 27.5.% for Clone 2), and peroxynitrite (24.3% for Clone 1; 43.9% for Clone 2), but not to exogenous application of HNE in culture medium. GST transfectants treated with 1,1,4-tris (acetyloxy)nonane, a nontoxic derivative of HNE that is degraded to HNE intracellularly, demonstrated a statistically significant (p <.05) increase in viability in a dose-dependent manner compared with SY5Y cells transfected with vector alone. These results suggest that overexpression of GST increases resistance to endogenous HNE induced by oxidative stress or released in the degradation of 1,1,4-tris (acetyloxy)nonane, but not to exogenous application of HNE. PMID- 11425493 TI - Loss of apoB-100 secondary structure and conformation in hydroperoxide rich, electronegative LDL(-). AB - A subpopulation of low-density lipoproteins (LDL) is present in human plasma that contains lipid hydroperoxides and is more negatively charged (LDL(-)) than normal native LDL. By circular dichroism and tryptophan lifetime measurements we found that apoB-100 secondary structure is markedly decreased and its conformation is severely altered in LDL(-). The low tryptophan fluorescence intensity confirms the oxidative degradation of the lipoprotein, and the very long lifetime value of one of its decay components indicates a low polarity environment for the remaining unbleached residues. Either a peculiar folding or, most likely, a sinking of the apoB-100 into the lipid core can account for the observed long lifetime component. Oxidation in vitro produces a similar unfolding of the apolipoprotein but the lifetime of tryptophan fluorescence is shifted to lower values, indicating that the denatured apoprotein remains at the hydrophilic surface of the lipoprotein particle. A disordering and an increased polarity of the LDL(-) surface lipids was demonstrated by measuring the generalized polarization of 2-dimethylamino-6-lauroylnaphthalene (Laurdan). The looser monolayer packing apparently favors the new conformation of apoB-100 and its sinking into a more hydrophobic environment, possibly accounting for it reduced receptor binding properties. PMID- 11425494 TI - Gene expression analysis of an H(2)O(2)-resistant lens epithelial cell line. AB - Gene expression patterns were examined in lens epithelial cells conditioned to grow in 125 microM hydrogen peroxide in order to define the protective mechanisms that may be involved in survival during oxidative stress. RNA was extracted from normal and hydrogen peroxide-resistant alphaTN4 mouse lens epithelial cells. Gene expression was evaluated using Differential Display (DD) and RT-PCR. Upregulation of mRNAs for antioxidant and cellular defense enzymes was observed. The highest elevation detected was a 14-fold increase in catalase in the hydrogen peroxide resistant cells. Glutathione peroxidase, ferritin, and alphaB-crystallin were upregulated 2-fold, and reticulocalbin was upregulated 6-fold in the resistant cells. alphaA-crystallin was downregulated 5-fold, while aldose reductase and mitochondrial gene products were unchanged. Thus, in the alphaTN4 mouse lens cell line, long-term exposure to high levels of hydrogen peroxide elicited an upregulation of transcripts for enzymes involved in hydrogen peroxide degradation, metal binding, and chaperone function. Since mitochondrial gene transcription is sensitive to hydrogen peroxide, the presence of normal levels of mitochondrial transcripts, in this study, demonstrates the effectiveness of the antioxidant defense systems. PMID- 11425495 TI - Thiyl radicals abstract hydrogen atoms from carbohydrates: reactivity and selectivity. AB - Free radical damage of DNA is a well-known process affecting biological tissue under conditions of oxidative stress. Though carbohydrate-derived radicals are generally "repaired" by hydrogen transfer from thiols, the reverse possibility, namely hydrogen abstraction by thiyl radicals from carbohydrates, exists. The biological relevance of this process has been discussed controversially, especially because of the lack of rate constants. Therefore, we have measured rate constants for the hydrogen transfer reaction between thiyl radicals from cysteine and selected carbohydrates, 2-deoxy-D-ribose (dRib), 2-deoxy-D-glucose (dGls), alpha-D-glucose (Gls), and inositol (Ino). Rate constants are on the order of 10(4) M(-1)s(-1), with the highest average value for dRib, (2.7 +/- 1.0) x 10(4) M(-1)s(-1), and the lowest average value for dGls, (1.6 +/- 0.2) x 10(4) M(-1)s(-1), based on two ways of kinetic analysis, standard competition kinetics and stochastic simulation of the experimental results, respectively. In general, thiyl radicals attack preferentially the C(1)-H bond of the carbohydrates, to an extent of ca. 72% in dRib and 90% in dGls. Kinetic measurements were possible through a specifically designed competition system measuring the reaction of thiyl radicals with either the C-H bonds of the carbohydrates or the C(alpha)-H bond of cysteine under conditions where the extent of other competitive reactions of the thiyl radicals were minimized. PMID- 11425496 TI - Oxidative DNA damage in cultured cells and rat lungs by carcinogenic nickel compounds. AB - DNA damage in cultured cells and in lungs of rats induced by nickel compounds was investigated to clarify the mechanism of nickel carcinogenesis. DNA strand breaks in cultured cells exposed to nickel compounds were measured by using a pulsed field gel electrophoresis technique. Among nickel compounds (Ni(3)S(2), NiO (black), NiO (green), and NiSO(4)), only Ni(3)S(2), which is highly carcinogenic, induced lesions of both double- and single-stranded DNA in cultured human cells (Raji and HeLa cells). Treatment of cultured HeLa cells with Ni(3)S(2) (10 microg/ml) induced a 1.5-fold increase in 8-hydroxy-2'-deoxyguanosine (8-OH-dG) compared with control, whereas NiO (black), NiO (green), and NiSO(4) did not enhance the generation of 8-OH-dG. Intratracheal instillation of Ni(3)S(2), NiO(black), and NiO(green) to Wistar rats increased 8-OH-dG in the lungs significantly. NiSO(4) induced a smaller but significant increase in 8-OH-dG. Histological studies showed that all the nickel compounds used induced inflammation in lungs of the rats. Nitric oxide (NO) generation in phagocytic cells induced by Ni(3)S(2), NiO(black), and NiO(green) was examined using macrophage cell line RAW 264.7 cells. NO generation in RAW 264.7 cells stimulated with lipopolysaccharide was enhanced by all nickel particles. Two mechanisms for nickel-induced oxidative DNA damage have been proposed as follows: all the nickel compounds used induced indirect damage through inflammation, and Ni(3)S(2) also showed direct oxidative DNA damage through H(2)O(2) formation. This double action may explain relatively high carcinogenic risk of Ni(3)S(2). PMID- 11425497 TI - Recycling of the ascorbate free radical by human erythrocyte membranes. AB - Reduction of the ascorbate free radical (AFR) at the plasma membrane provides an efficient mechanism to preserve the vitamin in a location where it can recycle alpha-tocopherol and thus prevent lipid peroxidation. Erythrocyte ghost membranes have been shown to oxidize NADH in the presence of the AFR. We report that this activity derives from an AFR reductase because it spares ascorbate from oxidation by ascorbate oxidase, and because ghost membranes decrease steady-state concentrations of the AFR in a protein- and NADH-dependent manner. The AFR reductase has a high apparent affinity for both NADH and the AFR (< 2 microM). When measured in open ghosts, the reductase is comprised of an inner membrane activity (both substrate sites on the cytosolic membrane face) and a trans membrane activity that mediates extracellular AFR reduction using intracellular NADH. However, the trans-membrane activity constitutes only about 12% of the total measured in ghosts. Ghost AFR reductase activity can also be differentiated from NADH-dependent ferricyanide reductase(s) by its sensitivity to the detergent Triton X-100 and insensitivity to enzymatic digestion with cathepsin D. This NADH dependent AFR reductase could serve to recycle ascorbic acid at a crucial site on the inner face of the plasma membrane. PMID- 11425498 TI - Effect of alpha-lipoic acid on vascular responses and nociception in diabetic rats. AB - Oxidative stress contributes to the vascular and neurological complications of diabetes mellitus. The aim was to evaluate the effects of treatment with the radical scavenger and transition metal chelator, alpha-lipoic acid, on endothelium-dependent relaxation of the mesenteric vasculature and on superior cervical ganglion blood flow in 8 week streptozotocin-induced diabetic rats. alpha-Lipoic acid effects on small nerve fiber-mediated nociception were also monitored. For the in vitro phenylephrine-precontracted mesenteric vascular bed, diabetes caused a 31% deficit in maximum endothelium-dependent relaxation to acetylcholine, and a 4-fold reduction in sensitivity. alpha-Lipoic acid gave 85% protection against these defects. Acetylcholine responses are mediated by nitric oxide and endothelium-derived hyperpolarizing factor: isolation of the latter by nitric oxide synthase blockade revealed a 74% diabetic deficit that was halved by alpha-lipoic acid. Superior cervical ganglion blood flow, 52% reduced by diabetes, was dose-dependently restored by alpha-lipoic acid (ED(50), 44 mg/kg/d). Diabetic rats exhibited mechanical and thermal hyperalgesia, which were abolished by alpha-lipoic acid treatment. Thus, diabetes impairs nitric oxide and endothelium-derived hyperpolarizing factor-mediated vasodilation. This contributes to reduced neural perfusion, and may be responsible for altered nociceptive function. The effect of alpha-lipoic acid strongly implicates oxidative stress in these events and suggests a potential therapeutic approach. PMID- 11425499 TI - Continuous phencyclidine treatment induces schizophrenia-like hyperreactivity of striatal dopamine release. AB - Functional dopaminergic hyperactivity is a key feature of schizophrenia. Recent in vivo imaging studies have demonstrated greater striatal dopamine release in response to amphetamine challenge in schizophrenia subjects than in normal controls. N-methyl-D-aspartate (NMDA) receptors are known to play a prominent role in regulation of striatal dopamine release. In humans, NMDA antagonists induce a psychotic state that closely resembles schizophrenia. The present study investigates the degree to which chronic continuous administration of the NMDA antagonist phencyclidine (PCP) induces schizophrenia-like hyperreactivity of striatal dopamine release to amphetamine in rodents. Rats were treated with 10 or 15 mg/kg/d PCP for two weeks by osmotic minipump, and striatal dopamine release to amphetamine challenge (1 mg/kg) was monitored by microdialysis. PCP-treated rats showed significant enhancement in amphetamine-induced dopamine release, along with significantly enhanced locomotor activity. These findings support the concept that NMDA receptor dysfunction may contribute to dopaminergic dysfunction in schizophrenia. PMID- 11425500 TI - Sequential regional cerebral blood flow brain scans using PET with H2(15)O demonstrate ketamine actions in CNS dynamically. AB - The aim of this study was to examine the potential of serial rCBF studies to directly characterize the regional effects and dynamic time course of the centrally active drug ketamine. The value of a broader application of this technique to other neurally active drugs to characterize the pharmacodynamics of CNS compounds is suggested by these data. Thirteen normal subjects received a 0.3 mg/kg intravenous dose of ketamine over 60 seconds; ten other individuals received placebo in the same manner. For each subject, three baseline PET rCBF scans and seven sequential post-ketamine scans at 10-minute intervals were obtained using H(2)(15)O water. SPM techniques were employed to identify the maxima of any cluster significant by spatial extent analysis at any post-ketamine time point between 0 and 36 min. These extremes from the ketamine group, were identified in placebo scans similarly and grown to a 6x6x12 mm voxel set. The average rCBF values of the ketamine-defined clusters were determined in the drug and placebo conditions at all time points. rCBF across time was plotted for each cluster and compared between drug and placebo. Area under the curve (AUC) was calculated between baseline and 36 minutes. The kinetic characteristics of the ketamine-induced rCBF curves were compared to induced behaviors in each maxima. Ketamine produced distinct patterns of rCBF change over time in different brain regions; maxima within an anatomically defined region responded similarly. Ketamine induced rCBF activations in anterior cingulate, medial frontal and inferior frontal cortices. All maxima with a relative flow reduction with ketamine were in the cerebellum. The pattern of all activations and suppressions was monophasic with the peak changes at 6-16 minutes. In preliminary analysis, individual C(max) and AUC of maxima in the anterior cingulate/medial frontal region tended to correlate with the mild psychotomimetic action of ketamine; whereas, there was no tendency toward correlation with this psychological change in cerebellar maxima. The direct action of a centrally active drug can be assessed regionally and dynamically in brain using rCBF and a scan sequence optimally timed to complement the drug's time course. Ketamine pharmacodynamic response can be related to concurrent behavioral changes, tending to link the behavior with a brain region. This experimental design provides direct characterization of drug action in the CNS in ways heretofore unavailable. PMID- 11425501 TI - Effects of single and repeated phencyclidine administration on the expression of metabotropic glutamate receptor subtype mRNAs in rat brain. AB - Recent animal studies regarding phencyclidine (PCP), which induces psychotic symptoms in humans, have suggested that group II metabotropic glutamate receptors (mGluRs) represent a novel target for the treatment of PCP psychosis. In the present study, we used in situ hybridization to investigate the gene expressions of the mGluR 1-5 subtypes following single and repeated administration of PCP in rats. A single administration of PCP (7.5mg/kg, i.p.,) resulted in a significant decrease in the mGluR5 mRNA expression of group I mGluR in the subcortical regions (thalamus (-15%), central gray (-23%), inferior colliculus (-23%), and nucleus accumbens (-10%)) and hippocampal formation (CA1 (-14%), CA2 (-15%), CA3 (-18%), and dentate gyrus (-18%)). After repeated PCP administration for 14 days, the mGluR2 mRNA expression of group II mGluR in the anterior cingulate cortex ( 23%) and the mGluR4 mRNA expression of group III mGluR in the cortical regions (parietal (-11%), temporal (-13%) and entorhinal cortices (-18%)), the caudate putamen (-12%), thalamus (-17%), and subiculum (-25%) were significantly decreased. These results indicate that PCP affects not only group II mGluR but also group I and III of mGluR, and it is of particular interest that mGluR2 subtype is involved in a development of behavioral abnormality following repeated PCP administration. Single and repeated administrations of PCP independently regulate the expression of mGluR subtypes of mRNA in the brain. PMID- 11425502 TI - Extinction of cocaine self-administration produces a differential time-related regulation of proenkephalin gene expression in rat brain. AB - The purpose of this study was to examine the time course effects of extinction of cocaine self-administration behavior on proenkephalin (PENK) gene expression in caudate-putamen nucleus (ST), nucleus accumbens (Acc), olfactory tubercle (Tu), piriform cortex (Pir), ventromedial hypothalamic nucleus (VMN), and central amygdala (Ce) as measured by in situ hybridization histochemistry. Seventy-two littermate male Lewis rats were randomly assigned in triads to one of three conditions: (1) contingent intravenous self-administration of 1 mg/kg/injection of cocaine (CONT); (2) noncontingent injections of either 1 mg/kg/injection of cocaine (NONCONT); or (3) saline yoked (SALINE) to the intake of the self administering subject. The self-administering rats were trained to self administer cocaine under a FR5 schedule of reinforcement for a minimum of 3 weeks. After stable baseline levels of drug intake had been reached, saline was substituted for drug. Following this first extinction period, cocaine self administration was reinstated for an additional period of 2 weeks. Immediately after cessation of the last session of cocaine self-administration (day 0) and 1 , 5-, and 10-day after the second extinction period, animal brains in each triad were removed to be processed for in situ hybridization. PENK mRNA levels were significantly higher in the cocaine groups when compared with SALINE group in the ST, Acc, Pir, and Tu regions on days 0, 1, 5, and 10 of the extinction and lower in the Ce region of CONT group when compared to NONCONT and SALINE groups on days 1, 5, and 10 of the extinction period. In the VMN nucleus, PENK mRNA content in CONT group versus NONCONT and SALINE groups was also lower, but there were statistically significant differences only on day 5. These results suggest that changes in PENK gene expression after contingent cocaine administration might be involved in cocaine withdrawal states. PMID- 11425503 TI - Pre-exposure to (+/-)3,4-methylenedioxy-methamphetamine (MDMA) facilitates acquisition of intravenous cocaine self-administration in rats. AB - Pre-exposure to (+/-)3,4-methylenedioxymeth-amphetamine (MDMA) elevates locomotor activity and extracellular dopamine levels in the nucleus accumbens following a cocaine challenge. The present study determined whether MDMA-induced sensitization to the effects of cocaine could be demonstrated in rats self administering cocaine. Three groups of rats were treated with saline (Sal), 5 mg/kg MDMA (once per day for 10 days; MDMA-5) or 20 mg/kg MDMA (twice per day for 4 days; MDMA-20). Subsequently, spontaneous acquisition of cocaine self administration was measured in 12 daily 2-h sessions. During these test sessions, two response levers were present. Responses on one lever delivered infusions of 0.1 mg of cocaine; responses on the other lever had no programmed consequences. Group Sal showed a weak preference for the active lever; whereas, group MDMA-20 exhibited a stronger active lever preference. By day 12, the MDMA-20 group earned approximately twice the number of cocaine infusions as those in group SAL. At this time point, more than twice as many rats in group MDMA-20 were taking a minimum of 10 infusions per session, as compared to group Sal. Rats in group MDMA 5 did not seem to differ from group Sal in terms of lever discrimination, number of cocaine infusions, and percentage of rats obtaining a criterion of 10 infusions. These results indicate that pre-exposure to a high dose of MDMA may facilitate acquisition of cocaine self-administration. This dosing regimen of MDMA is likely to release DA and to be neurotoxic to 5-HT neurons. Either or both of these mechanisms could contribute to the ability of MDMA to facilitate cocaine self-administration. PMID- 11425504 TI - Behavioral expression of opiate withdrawal is altered after prefrontocortical dopamine depletion in rats: monoaminergic correlates. AB - The objective of this study was to establish the effects of prefrontocortical dopamine depletion on opiate withdrawal and prefrontocortical neurochemical changes elicited by morphine dependence and withdrawal. The dopaminergic content was also measured in the nucleus accumbens during withdrawal, in order to detect reactive changes induced by prefrontocortical lesion. Withdrawal was induced by naloxone in morphine-dependent rats. Monoamine levels were analyzed post-mortem by high performance liquid cromatography. The results showed that chronic morphine dependence did not modify basal levels of monoamines in sham rats, revealing neuroadaptation of prefrontocortical dopamine, noradrenaline and serotonin systems to chronic morphine. The neuroadaptive phenomenon remained after prefrontocortical lesion (> 79% dopamine depletion). On the other hand, a strong increase of dopamine, noradrenaline, and serotonin contents in the medial prefrontal cortex of sham rats was detected during opiate withdrawal. However, in lesioned rats, the increase of prefrontocortical dopamine and serotonin content, but not that of noradrenaline, was much lower. In the nucleus accumbens, prefrontocortical lesion reactively enhanced the dopaminergic tone and, although opiate withdrawal reduced dopaminergic activity in both sham and lesioned rats, this reduction was less intense in the latter group. At a behavioral level, some symptoms of physical opiate withdrawal were exacerbated in lesioned rats (writhing, mastication, teeth-chattering, global score) and exploration was reduced. The findings hence indicate that: (i) prefrontocortical monoaminergic changes play a role in the behavioral expression of opiate withdrawal; (ii) the severity of some withdrawal signs are related to the dopaminergic and serotonergic tone of the medial prefrontal cortex rather than to the noradrenergic one, and (iii) an inverse relationship between mesocortical and mesolimbic dopaminergic systems exists. PMID- 11425505 TI - A simple method to measure baseline occupancy of neostriatal dopamine D2 receptors by dopamine in vivo in healthy subjects. AB - The effect of endogenous dopamine (DA) on measurement of neostriatal DA D(2) receptor binding potential (D(2)RBP) in vivo was evaluated with positron emission tomography (PET) and the radiotracer [11C]raclopride by comparing the D(2)RBP before and after acute DA depletion. DA depletion was achieved by per-oral administration of 4.5 g alpha-methyl-para-tyrosine (AMPT) given in 25 h. Six healthy subjects completed the protocol. The AMPT treatment increased D(2)RBP significantly from 3.11 +/- 0.25 to 3.68 +/- 0.23 and decreased plasma levels of the DA metabolite homovanillic acid by 71 +/- 11% and levels of the norepinephrine metabolite 3-methoxy-4-hydroxyphenethyleneglycol by 53 +/- 7%. Increase in D(2)RBP correlated with decrease in attentiveness and with increase in errors of commission from Conners' Continuous Performance Test. On AMPT, a significant decrease in subjective happiness scores was observed. The results imply that a noninvasive [11C]raclopride PET protocol coupled with relatively brief administration of a rather low total dose of AMPT resulted in measurable acute DA depletion that might provide estimates of synaptic neostriatal DA concentration. PMID- 11425506 TI - Effect of olanzapine on functional responses from sensitized D1-dopamine receptors in rats with neonatal dopamine loss. AB - Previous work has suggested that the therapeutic efficacy of olanzapine might be partially dependent on action at the D(1)-dopamine (DA) receptor site. Because early DA loss can lead to supersensitive D(1)-DA receptors, effects of olanzapine were investigated in adult rats given lesions to DA-containing neurons as neonates. In these animals, locomotor effects of SKF-38393 (a D(1)-DA agonist) were attenuated by olanzapine, but at doses (5 and 10 mg/kg) that decreased activity when given alone. Olanzapine prevented induction of striatal Fos protein by SKF-38393 and partially attenuated the long-term "priming" effect of repeated SKF-38393 treatment. Olanzapine also antagonized the stimulant effects of quinpirole (a D(2)-type DA agonist) in animals lesioned as young adults, at doses lower than those necessary to antagonize SKF-38393-induced activity. In addition, olanzapine antagonized apomorphine-induced self-injurious behavior in neonate lesioned rats in a dose-related fashion. Attenuation of self-injury in this animal model suggests that olanzapine should be tested against this symptom in patient populations. PMID- 11425507 TI - Repeated exposure to rewarding brain stimulation downregulates GluR1 expression in the ventral tegmental area. AB - There is considerable evidence that drug reward and brain stimulation reward (BSR) share common neural substrates. Although it is known that exposure to drugs of abuse causes a variety of molecular changes in brain reward systems, little is known about the molecular consequences of BSR. We report that repeated exposure to rewarding stimulation of the medial forebrain bundle (MFB) selectively decreases expression of GluR1 (an AMPA receptor subunit) in the VTA, without effect on expression of several other proteins (GluR2, NMDAR1, tyrosine hydroxylase). This effect of BSR on GluR1 expression is opposite of that caused by intermittent exposure to cocaine and morphine, which are known to elevate GluR1 expression in the VTA. Considering that elevated GluR1 expression in the VTA has been associated with increased sensitivity to drug reward, the finding that BSR and drugs of abuse have opposite effects on GluR1 expression in this region may provide an explanation for why the reward-related effects of many drugs (cocaine, morphine, amphetamine, PCP, nicotine) do not sensitize with repeated testing in BSR procedures that quantify reward strength. PMID- 11425508 TI - Estrogen-like activity of tamoxifen and raloxifene on NMDA receptor binding and expression of its subunits in rat brain. AB - Hormonal specificity of modulation of N-methyl- D-aspartate (NMDA) receptors was investigated by comparing the effects of estradiol with tamoxifen or raloxifene, which display different responses in breast, bone, and uterus. Two weeks ovariectomy in rats decreased uterine weight, which was prevented by a two-week estradiol treatment; tamoxifen and raloxifene had weaker uterine stimulation than estradiol. Ovariectomy in rats decreased L-[3H]glutamate specific binding to NMDA receptors in CA1 and dentate gyrus but not CA2/3 regions of hippocampus and was without effect in cortex, striatum, nucleus accumbens, and olfactory tubercle. [3H]Ro 25-6981 (an NMDA antagonist selective for NR1/NR2B assembly) specific binding and mRNA levels of NMDA receptor subunits 1 and 2B decreased in CA1 after ovariectomy. Estradiol, tamoxifen, and raloxifene decreased L-[3H]glutamate specific binding to NMDA receptors and [3H]Ro 25-6981 specific binding in cortical area of ovariectomized rats and prevented the decrease of [3H]glutamate specific binding to NMDA receptors in CA1 and dentate gyrus, as well as [3H]Ro 25 6981 specific binding in CA1. Estradiol prevented the decrease of NMDA receptor subunits 1 and 2B mRNA levels in CA1 only; tamoxifen and raloxifene prevented the decrease of NMDA receptor subunit 1 mRNA levels in CA1. No effect of ovariectomy or treatments on L-[3H]CGP 39653 (an NMDA antagonist selective for NR1/NR2A assembly) specific binding and NMDA receptor subunit 2A mRNA levels was observed in all brain regions assayed. Our results showed brain regional and subunits specific agonist estrogenic activity of tamoxifen and raloxifene on NMDA receptors. PMID- 11425509 TI - Effects of age and gender on the expression of brain-derived neurotrophic factor mRNA in rat retrosplenial cortex following administration of dizocilpine. AB - Using in situ hybridization, we studied the effects of age and gender on the expression of brain-derived neurotrophic factor (BDNF) mRNA and heat shock protein hsp-70 mRNA in the rat retrosplenial cortex following administration of the noncompetitive NMDA receptor antagonist (+)-MK-801 (dizocilpine). Male and female Sprague-Dawley rats (5 weeks, 12 weeks, or 10 months old) were given a single intraperitoneal injection of saline (1 ml/kg) or dizocilpine (0.3, 1.0, or 3.0 mg/kg). No expression of BDNF mRNA and hsp-70 mRNA was detected in the rat retrosplenial cortex after administration of saline (1 ml/kg, IP). Administration of dizocilpine (0.3, 1.0, or 3.0 mg/kg, IP) caused a marked induction of BDNF mRNA and hsp-70 mRNA in the retrosplenial cortex of male and female rats, in a dose-dependent manner. Female rats were more sensitive to the induction of BDNF mRNA and hsp-70 mRNA in the retrosplenial cortex by dizocilpine as compared to male rats. It was also found that adult (12 weeks old) and aged (10 months old) rats were more sensitive to the induction of hsp-70 mRNA and BDNF mRNA in the retrosplenial cortex by dizocilpine as compared to young (5 weeks old) rats. These results suggest that the age and gender differences observed in the expression of BDNF mRNA and hsp-70 mRNA in the retrosplenial cortex by dizocilpine may be associated with the differences in dizocilpine-induced neurotoxicity observed with gender and age within the same region. PMID- 11425510 TI - Twenty-four-hour ACTH and cortisol pulsatility in depressed women. AB - Increased plasma cortisol in patients with major depression is a well documented finding, although it is present in only 25-30% of subjects with major depression. However, ACTH and cortisol are secreted in a pulsatile manner, so it is unclear if increased ACTH secretion occurs in depression and if there are changes in the pulsatile components of ACTH secretion. Ten-minute sampling for ACTH and cortisol was performed for 24 hr in 25 premenopausal depressed women, whose age and menstrual cycle day matched control women. As a group, the depressed women demonstrated a trend to increase cortisol secretion (p = 0.089). There was no difference in mean cortisol between the patient group as a whole (8.36 +/- 2.9 microg/dl) and those patients meeting criteria for atypical depression (8.38 +/- 1.9 microg/dl), but patients meeting criteria for endogenous showed increased cortisol (12.17 +/- 4 microg/dl) Mean ACTH was not significantly different between patients and controls. Pulse analyses revealed similar number of secretory events and similar amplitudes for cortisol secretory bursts in patients and controls. The baseline component area under the curve of cortisol secretion was increased at a trend level (p =.064) in depressed patients, and the baseline AUC for ACTH was significantly increased in depressed patients (p =.045). No differences were found in pulsatile components of ACTH secretion between patients and matched controls. Harmonic analyses indicated no significant differences between patients and controls on any detected rhythm for ACTH or cortisol. These data suggest that the pulsatile and circadian components of the HPA axis are normal in premenopausal depressed women and that only 24% of depressed women demonstrate hypercortisolemia. PMID- 11425511 TI - The effect of tryptophan on social interaction in everyday life: a placebo controlled study. AB - In monkeys increasing serotonin function enhances affiliative interactions and promotes the acquisition of dominance. To examine whether similar effects occur in humans, we treated 98 subjects for 12 days with the serotonin precursor tryptophan (1g TID) and for 12 days with placebo in a double-blind, cross over study. Agreeableness/quarrelsomeness and dominance/submission were measured using an event-contingent method, in which subjects reported on various behaviors during important social interactions throughout their day. Tryptophan decreased quarrelsome behavior, but only when placebo was given first, suggesting that a decrease in quarrelsomeness when tryptophan was given first may have carried over into the subsequent placebo period. Tryptophan increased dominant behavior, an effect that was independent of the order of treatment, the broad social context, and the subject's and partner's sex. Our results suggest that serotonin may enhance dominance in humans, as in monkeys, and illustrate the advantages of the event contingent methodology in studying the associations between biology and human social interaction. PMID- 11425512 TI - Translesion synthesis by the UmuC family of DNA polymerases. AB - Translesion synthesis is an important cellular mechanism to overcome replication blockage by DNA damage. To copy damaged DNA templates during replication, specialized DNA polymerases are required. Translesion synthesis can be error-free or error-prone. From E. coli to humans, error-prone translesion synthesis constitutes a major mechanism of DNA damage-induced mutagenesis. As a response to DNA damage during replication, translesion synthesis contributes to cell survival and induced mutagenesis. During 1999-2000, the UmuC superfamily had emerged, which consists of the following prototypic members: the E. coli UmuC, the E. coli DinB, the yeast Rad30, the human RAD30B, and the yeast Rev1. The corresponding biochemical activities are DNA polymerases V, IV, eta, iota, and dCMP transferase, respectively. Recent studies of the UmuC superfamily are summarized and evidence is presented suggesting that this family of DNA polymerases is involved in translesion DNA synthesis. PMID- 11425513 TI - Molecular mechanisms of DNA mismatch repair. AB - DNA mismatch repair (MMR) safeguards the integrity of the genome. In its role in postreplicative repair, this repair pathway corrects base-base and insertion/deletion (I/D) mismatches that have escaped the proofreading function of replicative polymerases. In its absence, cells assume a mutator phenotype in which the rate of spontaneous mutation is greatly elevated. The discovery that defects in mismatch repair segregate with certain cancer predisposition syndromes highlights its essential role in mutation avoidance. Recently, three-dimensional structures of MutS, a key repair protein that recognizes mismatches, have been determined by X-ray crystallography. This article provides an overview of the structural features of MutS proteins and discusses how the structural data together with biochemical and genetic studies reveal new insights into the molecular mechanisms of mismatch repair. PMID- 11425514 TI - The p48 subunit of the damaged-DNA binding protein DDB associates with the CBP/p300 family of histone acetyltransferase. AB - DDB has been implicated in DNA repair as well as transcription. Mutations in DDB have been correlated with the repair-deficiency disease, xeroderma pigmentosum group E (XP-E). The XP-E cells exhibit deficiencies in global genomic repair, suggesting a role for DDB in that process. DDB also possesses a transcription stimulatory activity. We showed that DDB could function as a transcriptional partner of E2F1. But the mechanism by which DDB stimulates E2F-regulated transcription or carry out its DNA repair function is not understood. To investigate the mechanisms, we looked for nuclear proteins that interact with DDB. Here we show that DDB associates with the CBP/p300 family of proteins, in vivo and in vitro. We suggest that DDB participates in global genomic repair by recruiting CBP/p300 to the damaged-chromatin. It is possible that the histone acetyltransferase activities of the CBP/p300 proteins induce chromatin remodeling at the damaged-sites to allow recruitment of the repair complexes. The observation offers insights into both transcription and repair functions of DDB. PMID- 11425515 TI - Repair activities of 8-oxoguanine DNA glycosylase from Archaeoglobus fulgidus, a hyperthermophilic archaeon. AB - Oxidative DNA damage is caused by reactive oxygen species formed in cells as by products of aerobic metabolism or of oxidative stress. The 8-oxoguanine (8-oxoG) DNA glycosylase from Archaeoglobus fulgidus (Afogg), which excises an oxidatively damaged form of guanine, was overproduced in Escherichia coli, purified and characterized. A. fulgidus is a sulfate-reducing archaeon, which grows at between 60 and 95 degrees C, with an optimum growth at 83 degrees C. The Afogg enzyme has both DNA glycosylase and apurinic/apyrimidinic (AP) lyase activities, with the latter proceeding through a Schiff base intermediate. As expected for a protein from a hyperthermophilic organism, the enzyme activity is optimal near pH 8.5 and 60 degrees C, denaturing at 80 degrees C, and is thermally stable at high levels of salt (500mM). The Afogg protein efficiently cleaves oligomers containing 8 oxoG:C and 8-oxoG:G base pairs, and is less effective on oligomers containing 8 oxoG:T and 8-oxoG:A mispairs. While the catalytic action mechanism of Afogg protein is likely similar to the human Ogg1 (hOgg1), the DNA recognition mechanism and the basis for 8-oxoG substrate specificity of Afogg differ from that of hOgg. PMID- 11425516 TI - Nucleotide excision repair gene expression in the rat conceptus during organogenesis. AB - DNA repair may be a determinant of the susceptibility of the conceptus to DNA damaging teratogens. The nucleotide excision repair (NER) pathway repairs a substantial amount of chemically induced DNA damage. The goals of this study were to assess the coordinate expression of NER genes in the midorganogenesis-stage rat conceptus and determine the consequences of exposure to the genotoxic teratogen, 4-hydroperoxycyclophosphamide (4-OOHCPA), on NER gene expression. Most NER genes were expressed at low levels in both yolk sac and embryo on gestational day (GD) 10, with the exception of XPD, XPE and PCNA. No significant alterations in gene expression occurred between GDs 10 and 11; in the yolk sac XPB expression increased on GD12 compared to either GD10 or 11. In the embryo, XPE expression increased between GDs 10 and 12, while hHR23B, XPB, ERCC1, and DNA polymerase epsilon expression increased on GD12 relative to both GDs 10 and 11. Contrary to gene expression data, XPB protein was found at high levels and XPD at low levels in GDs 10-12 embryos and yolk sacs. Mirroring gene expression, high levels of PCNA protein were found in both tissues; XPA protein levels were minimal in yolk sac from GDs 10-12 but increased in the embryo from moderate on GD10 to high on GD12. Therefore, NER gene expression during organogenesis was regulated in a developmental stage- and tissue-specific manner. Exposure of the conceptus to a teratogen, 4-OOHCPA, induced malformations without affecting NER transcript levels. Thus, NER gene expression in the conceptus was unresponsive to regulation by DNA alkylation. PMID- 11425517 TI - Spontaneous germline amplification and translocation of a transgene array. AB - The majority of the mammalian genome is thought to be relatively stable throughout and between generations. There are no developmentally programmed gene amplifications as seen in lower eukaryotes and prokaryotes, however a number of unscheduled gene amplifications have been documented. Apart from expansion of trinucleotide repeats and minisatellite DNA, which involve small DNA elements, other cases of gene or DNA amplifications in mammalian systems have been reported in tumor samples or permanent cell lines. The mechanisms underlying these amplifications remain unknown. Here, we report a spontaneous transgene amplification through the male germline which resulted in silencing of transgene expression. During routine screening one mouse, phenotypically negative for transgene expression, was found to have a transgene copy number much greater than that of the transgenic parent. Analysis of the transgene expansion revealed that the amplification in the new high copy transgenic line resulted in a copy number approximately 40-60 times the primary transgenic line copy number of 5-8 copies per haploid genome. Genetic breeding analysis suggested that this amplification was the result of insertion at only one integration site, that it was stable for at least two generations and that the site of insertion was different from the site at which the original 5-8 copy array had integrated. FISH analysis revealed that the new high copy array was on chromosome 7 F3/4 whereas the original low copy transgene array had been localised to chromosome 3E3. DNA methylation analysis revealed that the high copy transgene array was heavily methylated. The amplification of transgenes, although a rare event, may give insight into amplification of endogenous genes which can be associated with human disease. PMID- 11425518 TI - Deletion of the SRS2 gene suppresses elevated recombination and DNA damage sensitivity in rad5 and rad18 mutants of Saccharomyces cerevisiae. AB - The Saccharomyces cerevisiae genes RAD5, RAD18, and SRS2 are proposed to act in post-replicational repair of DNA damage. We have investigated the genetic interactions between mutations in these genes with respect to cell survival and ectopic gene conversion following treatment of logarithmic and early stationary cells with UV- and gamma-rays. We find that the genetic interaction between the rad5 and rad18 mutations depends on DNA damage type and position in the cell cycle at the time of treatment. Inactivation of SRS2 suppresses damage sensitivity both in rad5 and rad18 mutants, but only when treated in logarithmic phase. When irradiated in stationary phase, the srs2 mutation enhances the sensitivity of rad5 mutants, whereas it has no effect on rad18 mutants. Irrespective of the growth phase, the srs2 mutation reduces the frequency of damage-induced ectopic gene conversion in rad5 and rad18 mutants. In addition, we find that srs2 mutants exhibit reduced spontaneous and UV-induced sister chromatid recombination (SCR), whereas rad5 and rad18 mutants are proficient for SCR. We propose a model in which the Srs2 protein has pro-recombinogenic or anti recombinogenic activity, depending on the context of the DNA damage. PMID- 11425519 TI - Cellular effects of 5-formyluracil in DNA. AB - 5-Formyluracil is a major oxidation product of thymine, formed in DNA in yields comparable to that of 8-oxo-7,8-dihydroguanine by exposure to gamma-irradiation. Whereas the repair pathways for removal and the biological effects of persisting 8-oxo-7,8-dihydroguanine are much elucidated, much less attention has been paid to the cellular implications of 5-formyluracil in DNA. Here we review the present state of knowledge in this important area within research on oxidative DNA damage. PMID- 11425520 TI - Heterocyclic amine induced apoptotic response in the human lymphoblastoid cell line TK6 is linked to mismatch repair status. AB - The human lymphoblastoid cell, TK6, exhibited a dose-dependent cytotoxic and apoptotic response following treatment with the food borne heterocyclic amine, 2 amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Augmentation of the p53 protein and increases in p21-WAF1 levels were also observed. Comparison of the survival by clonogenic assays and the percentage of apoptotic cells (cells containing subG1 DNA or condensed nuclei) revealed that only 10-20% of the PhIP induced cell death could be attributed to apoptosis that occurred in the first 24h after treatment. MT1, a derivative of TK6 that contains mutations in both alleles of its hMSH6 gene and is mismatch repair deficient, showed a decreased apoptotic response. A significant increase (P<0.05) in apoptosis was observed in TK6 and not in MT1 following treatment with 2.5microg/ml PhIP. A five- to six fold increase and less than a two-fold increase in the fraction of apoptotic cells were observed in TK6 and MT1, respectively. Treatment with 5microg/ml PhIP resulted in significant increases in apoptosis (P<0.05) in TK6 and MT1. The percentages of apoptotic cells were, however, two- to three-fold higher in TK6 than in MT1. HCT116, a hMLH1 defective mismatch repair deficient colorectal carcinoma cell line, also exhibited lower PhIP-induced apoptosis than its mismatch repair proficient chromosome transfer cell line (HCT116+chr3) following PhIP treatment. These results show that PhIP-induced apoptosis is mediated through a mismatch repair dependent pathway. Accumulation of p53 in TK6 and MT1 were evident in samples taken 24h after PhIP treatment. Increases in p21-WAF1 were also observed in both cell lines confirming that the p53 was functional. The lower apoptotic response of MT1 but similar p53 accumulation in TK6 and MT1 suggest that the mismatch repair protein(s) are involved downstream of p53 or that PhIP-induced apoptosis is p53-independent. PMID- 11425522 TI - Enantioselective synthesis and complement inhibitory assay of A/B-ring partial analogues of oleanolic acid. AB - A series of oleanolic acid A/B-ring partial analogues was synthesized and tested for their complement inhibitory activity as well as cytotoxic properties. All target compounds and one intermediate exhibited moderate complement inhibitory potency. These compounds also showed cytotoxicity on malignant melanoma cell line, SK-MEL. PMID- 11425523 TI - Novel heteroaryl replacements of aromatic 3-tetrafluoroethoxy substituents in trifluoro-3-(tertiaryamino)-2-propanols as potent inhibitors of cholesteryl ester transfer protein. AB - A series of novel N,N-disubstituted trifluoro-3-amino-2-propanols has been prepared as potent inhibitors of cholesteryl ester transfer protein (CETP). Modifying the aromatic 3-tetrafluoroethoxy group in the lead molecule 1a with various heteroaryl moieties produced new 2-furyl analogues 2a,b with submicromolar potency in vitro. PMID- 11425524 TI - Dibasic inhibitors of human mast cell tryptase. Part 3: identification of a series of potent and selective inhibitors containing the benzamidine functionality. AB - A survey of charged groups and linkers for a series of symmetrical and unsymmetrical dibasic inhibitors is described, leading to several classes of potent and selective inhibitors. In particular, the benzamidine functionality was identified as the most potent charged group investigated. PMID- 11425525 TI - Sequence-selective peptide detection by small synthetic chemosensors selected from an encoded combinatorial chemosensor library. AB - Synthetic chemosensors hold great potential in many diagnostic applications. In this study, we describe the design and preparation of the first encoded combinatorial library of chemosensors for tripeptides. Subsequent screening of the library resulted in the discovery of novel chemosensors able to distinguish between random tripeptides. PMID- 11425526 TI - Quinoline-4-acetamides as sPLA(2) inhibitors. AB - Quinoline-4-acetamides were designed as potential phospholipase A(2) inhibitors by structural based method and synthesized. The chemical structures of the obtained compounds were confirmed by elemental analyses, 1H NMR and MS. Preliminary bioassay study shows that quinoline-4-acetamides display certain inhibition to sPLA(2). PMID- 11425527 TI - Expedited discovery of second generation NK-1 antagonists: identification of a nonbasic aryloxy substituent. AB - Solution-phase, parallel-synthesis techniques were used to optimize a series of nonbasic NK-1 antagonists, resulting in the identification of (R)-26, an orally bioavailable compound with subnanomolar potency. PMID- 11425528 TI - Biological evaluation of hepatitis C virus helicase inhibitors. AB - A small chemical library has been synthesized and assayed for inhibition of HCV helicase activity. This study provides the structure-activity relationship of the reported inhibitors, with emphasis placed on the aminophenylbenzimidazole moiety and the linkers. Our data highlight the importance of preserving the aminophenylbenzimidazole core and the hydrophobic linkers for biological activity. The development of a robust HCV helicase assay is also described. PMID- 11425529 TI - Anticonvulsant effects of new morphinan derivatives. AB - We synthesized a series of compounds that are modified in positions 3 and 17 of the morphinan ring system, with the intention of developing ideal anticonvulsant agents. We examined the effects of these compounds on kainic acid (KA)-induced seizures, and on locomotor patterns in rats. We found that compounds 5, 6, and 8 exhibit novel anticonvulsant effects, with negligible psychotropic effects. PMID- 11425530 TI - Nucleoside analogues as highly potent and selective inhibitors of herpes simplex virus thymidine kinase. AB - A series of carboxamide derivatives of 5'-amino-2',5'-dideoxy-5-ethyluridine has been prepared as inhibitors of HSV-TK (herpes simplex virus thymidine kinase). The most potent compounds were derived from xanthene, thioxanthene and dihydroanthracene carboxylic acids. The lead compounds show subnanomolar IC(50) values against HSV TKs. PMID- 11425531 TI - [3H]MFZ 2-12: a novel radioligand for the dopamine transporter. AB - In an effort to develop a tritiated dopamine transporter radioligand with higher affinity than the widely used [(3)H]WIN 35,428, we have synthesized [(3)H]2beta carbomethoxy-3beta-(3',4'-dichlorophenyl)tropane ([(3)H]MFZ 2-12). Unlabeled MFZ 2-12 and the N-demethylated intermediate (MFZ 2-13) inhibited dopamine uptake by the human dopamine transporter with IC(50)'s of 1.1 and 1.4nM, respectively. The N-nor-intermediate (MFZ 2-13) was treated with CT(3)I resulting in [(3)H]MFZ 2 12; S.A.=80 Ci/mmol). [(3)H]MFZ 2-12 reversibly bound with a K(D) of 2.8nM to human dopamine transporter expressed heterologously in EM4 cells. PMID- 11425532 TI - A simple method to obtain a covalent immobilized phospholipase A(2). AB - In the present work, we obtained an immobilized phospholipase A(2) system through covalent coupling by using an acrylic polymer Eupergit C as support. The immobilized enzyme from cobra venom (Naja naja naja) showed good retention activity and excellent stability. Both properties are of great importance for biomedical applications such as hypercholesterolemia treatments. PMID- 11425533 TI - Selective inhibition of Src SH2 by a novel thiol-targeting tricarbonyl-modified inhibitor and mechanistic analysis by (1)H/(13)C NMR spectroscopy. AB - Detailed analysis of Src SH2 binding by peptides containing a novel tricarbonyl modified pTyr moiety is described. We envisaged that Src SH2 selectivity might be obtained by exploiting the thiol group of Cys188 present in the pTyr binding pocket of the protein at the betaC3 position. Peptidyl as well as non-peptidyl compounds 1-4 possessing a 4-alpha,beta-diketoester-modified pTyr mimic exhibited micromolar affinity to Src SH2. Furthermore, these tricarbonyl compounds were selective for Src SH2 to the extent they showed no significant affinity for either Cys188Ser or Cys188Ala Src SH2 mutants. Upon closer examination of the binding of these tricarbonyls to Src SH2 using NMR of 13C-labeled compounds (6a, 6b, and 6c), we found that after the initial binding event the molecule disproportionated in a 'retro-Claisen' fashion to provide benzoic acid 16 and, following hydrolysis of the methyl ester 17, the hemiketal adduct of glyoxalic acid 18. PMID- 11425534 TI - Novel sulfonate derivatives: potent antimitotic agents. AB - The synthesis and biological evaluation of novel sulfonate analogues of E-7010 are reported. Several of the compounds are potent inhibitors of cell proliferation and tubulin polymerization. Importantly, these compounds are also active against P-glycoprotein positive (+) cancer cells, which are resistant to many other antitumor agents. PMID- 11425535 TI - Antimycobacterial activity of 9-sulfonylated/sulfenylated-6-mercaptopurine derivatives. AB - A series of 9-sulfonylated/sulfenylated-6-mercaptopurines has been prepared by reaction of 6-mercaptopurine with sulfonyl/sulfenyl halides. These compounds constitute a new class of potent antimycobacterial agents, possessing MIC values against Mycobacterium tuberculosis H37Rv in the range of 0.39-3.39 microg/mL, as well as appreciable activity against Mycobacterium avium. Furthermore, a compound of this small series exhibited good activity (MIC under 1 microg/mL) against several drug resistant strains of M. tuberculosis. PMID- 11425536 TI - Ethambutol analogues as potential antimycobacterial agents. AB - A range of new ethambutol analogues was synthesised and their inhibitory potencies were probed with Mycobacterium smegmatis. Interestingly, apparently even minor deviation from the structure of the parent compound resulted in reduced antimycobacterial activity. PMID- 11425537 TI - Synthesis of a novel C-10 spiro-epoxide of paclitaxel. AB - New analogues of paclitaxel (1a, active constituent of Taxol) were synthesized containing an epoxide at the C-10 position. The introduction of the epoxide was carried out by selective removal of the C10-acetate followed by protection of the C2'- and C7-hydroxyl groups. After oxidation to yield a ketone at the C10 position, this intermediate was reacted with dimethylsulfonium ylide. Deprotection and further manipulations provide the C10-spiro epoxide of paclitaxel (1b) and the corresponding C7-MOM ether (1c). PMID- 11425538 TI - Synthesis of substituted 5[H]phenanthridin-6-ones as potent poly(ADP ribose)polymerase-1 (PARP1) inhibitors. AB - 1-, 2-, 3-, 4-, 8-, or 10-Substituted 5(H)phenanthridin-6-ones were synthesized and found to be potent PARP1 inhibitors. Among the 28 compounds prepared, some showed not only low IC(50) values (compound 1b, 10 nM) but also desirable water solubility characteristics. These properties, which are superior to the common PARP1 inhibitors such as benzamides and isoquinolin-1-ones, are essential for potential therapeutic usage. The variety of compounds allows SAR analysis of favored substituents and substituted positions on 5(H)phenanthridin-6-one ring. PMID- 11425539 TI - Inhibition of serine proteases: activity of 1,3-diazetidine-2,4-diones. AB - The present work demonstrates that the 1,3-diazetidine-2,4-dione nucleus is effective as a scaffold of serine protease inhibitors. Compound 1 displayed high activity against human cathepsin G and alpha-chymotrypsin (0.39, 0.69 nM). Compound 6 exhibited 0.85 nM inhibition of human chymase. Compound 10 was a selective inhibitor against human neutrophil elastase. PMID- 11425540 TI - Total synthesis of human chymase inhibitor methyllinderone and structure- activity relationships of its derivatives. AB - Total synthesis of human chymase inhibitor methyllinderone has been achieved in only four steps with an overall yield of 21% from dimethyl squarate. We developed an efficient synthetic method for obtaining methyllinderone derivatives and found the active compound. In addition, we propose the inhibition mechanism of the active compound against human chymase using calculations. PMID- 11425541 TI - Antioxidant activity of water-soluble chitosan derivatives. AB - Water-soluble chitosan derivatives were prepared by graft copolymerization of maleic acid sodium onto hydroxypropyl chitosan and carboxymethyl chitosan sodium. Their scavenging activities against hydroxyl radical *OH were investigated by chemiluminescence technique. They exhibit IC(50) values ranging from 246 to 498 microg/mL, which should be attributed to their different contents of hydroxyl and amino groups and different substituting groups. PMID- 11425542 TI - Some 3-thioxo/alkylthio-1,2,4-triazoles with a substituted thiourea moiety as possible antimycobacterials. AB - A series of novel N-alkyl/aryl-N'-[4-(4-alkyl/aryl-2,4-dihydro-3H-1,2,4-triazole 3-thione-5-yl)phenyl]thioureas 1-19 and three S-alkylated representatives of the former, N-alkyl/aryl-N'-[4-(3-aralkylthio-4-alkyl/aryl-4H-1,2,4-triazole-5 yl)phenyl]thioureas 20-22, were synthesized and tested for antimycobacterial activity against Mycobacterium tuberculosis H37Rv as well as Mycobacterium fortuitum ATCC 6841 which is a rapid growing opportunistic pathogen. Compounds 4 and 9-11 were found to possess the same MIC value with that of Tobramycin against M. fortuitum ATCC 6841 whereas 1-3 and 21 had positive response against M. tuberculosis H37Rv at varying degrees. Compound 21 was identified as the most potent derivative of the 1-22 series by an MIC value of 6.25 microg/mL and selectivity index of 1.6. PMID- 11425543 TI - Biphenyls as surrogates of the steroidal backbone. Part 1: synthesis and estrogen receptor affinity of an original series of polysubstituted biphenyls. AB - In the course of a programme aimed at discovering new ligands of the estrogen receptor, we explored a series of substituted biphenyls. Their synthesis and binding affinity are described. PMID- 11425544 TI - Biphenyls as surrogates of the steroidal backbone. Part 2: discovery of a novel family of non-steroidal 5-alpha-reductase inhibitors. AB - A new family of non-steroidal 5-alpha-reductase inhibitors was designed by replacing the steroid skeleton of an inhibitor related to estrone by a biphenyl moiety. This hypothesis originated from the reported estrogenic activity of a few biphenyl compounds (see Part 1 of this paper; Lesuisse et al. Bioorg. Med. Chem. Lett. 2001, 11, 1709). Two compounds turned out to be potent type 2 5-alpha reductase inhibitors with IC(50)'s of inhibition in the nanomolar range. These are to our knowledge amongst the most potent non-steroidal 5-alpha-reductase inhibitors described to date. PMID- 11425545 TI - 3-Carboxamido analogues of morphine and naltrexone. synthesis and opioid receptor binding properties. AB - In response to the unexpectedly high affinity for opioid receptors observed in a novel series of cyclazocine analogues where the prototypic 8-OH was replaced by a carboxamido group, we have prepared the corresponding 3-CONH(2) analogues of morphine and naltrexone. High affinity (K(i)=34 and 1.7nM) for mu opioid receptors was seen, however, the new targets were 39- and 11-fold less potent than morphine and naltrexone, respectively. PMID- 11425546 TI - Substituted indole-5-carboxamides and -acetamides as potent nonpeptide GnRH receptor antagonists. AB - The 2-aryltryptamine class of GnRH receptor antagonists has been modified to incorporate carboxamide and acetamide substituents at the indole 5-position. With either a phenol or methanesulfonamide terminus on the N-aralkyl side chain, potent binding affinity to the GnRH receptor was achieved. A functional assay for GnRH antagonism was even more sensitive to structural modification and revealed a strong preference for branched tertiary amides. PMID- 11425547 TI - Potent nonpeptide GnRH receptor antagonists derived from substituted indole-5 carboxamides and -acetamides bearing a pyridine side-chain terminus. AB - A pyridine side-chain terminus has been incorporated into the indole-5 carboxamide and indole-5-acetamide series of GnRH antagonists. Potent activity was observed in binding and functional assays. Certain branched or cyclic tertiary amides were identified as preferred in each series. Alkylation of the side-chain secondary amine had generally unfavorable effects. Variations of the gem-dialkyl substituents in the indole-5-acetamide series were also investigated. PMID- 11425548 TI - Synthesis and nuclease stability of trilysyl dendrimer-oligodeoxyribonucleotide hybrids. AB - Hybrids of oligonucleotides and trilysyl-dendrimers with terminal acyl groups were prepared via solid-phase synthesis, including a DNA hexamer bearing an additional 3'-appendage. These were shown to be degraded more slowly by nuclease S1 than control strands, particularly at low pH, and, in one case, to form a duplex with a complementary strand whose melting point at pH 7 was higher than that of the control duplex. PMID- 11425549 TI - Isoindolines: a new series of potent and selective endothelin-A receptor antagonists. AB - 1,3-Disubstituted isoindolines have been discovered as a new class of potent functional ET(A) selective receptor antagonists through pharmacophore analysis of existing nonpeptide endothelin antagonists. The structure-activity relationships for both the trans and the cis series of isoindolines are discussed. PMID- 11425550 TI - Piperazinyl benzamidines: synthesis and affinity for the delta opioid receptor. AB - Piperazinyl benzamidines were prepared and found to bind to the rat delta (delta) opioid receptor. The most active compounds had a N,N-diethylcarboxamido group and a N-benzyl piperazine. The most potent among these was N,N-diethyl-4-[4 (phenylmethyl)-1-piperazinyl][2-(trifluoromethyl)phenyl]iminomethyl]benzamide (27) with a 1.22nM K(i) for the rat delta opioid receptor and ca. 1000 x selectivity relative to the mu opioid subtype. PMID- 11425551 TI - Promotion of radiation-induced formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine by nitro 5-deazaflavin derivatives. AB - 6-Nitro- and 8-nitro-5-deazaflavin derivatives have been found to enhance prominently the radiation-induced formation of 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodGuo) at the expense of formation of 2,6-diamino-4-hydroxy-5 formamidopyrimidine nucleosides (FapydGuo) both in deaerated and in N(2)O saturated aqueous 2'-deoxyguanosine solutions. The radiosensitizing capacity of a 9-nitro-5-deazflavin derivative was observed only in the N(2)O saturated aqueous solutions. PMID- 11425552 TI - Phenylcyanoguanidines as inhibitors of glucose-induced insulin secretion from beta cells. AB - 3,5-Disubstituted-phenylcyanoguanidines have been identified as activators of SUR1/Kir6.2 potassium channels and as potent inhibitors of insulin release from pancreatic beta cells in vitro. PMID- 11425553 TI - Significance of hydrogen bonding at the S(1)' subsite of calpain I. AB - alpha-Ketohydroxamates were synthesized as bioisosteres of alpha-ketoamides. The alpha-ketohydroxamates were generally more potent than the corresponding alpha ketoamides. The potency of the compounds suggests that hydrogen bonding and steric bulk of substituents on the nitrogen atom of the ketoamide moiety influence calpain inhibition. PMID- 11425554 TI - Inactivation of monoamine oxidase B by 1-phenylcyclopropylamine: mass spectral evidence for the flavin adduct. AB - Incubation of 1-phenylcyclopropylamine with bovine liver MAO (MAO B), followed by complete enzymatic digestion to single amino acid residues and subsequent analysis by on-line liquid chromatography-electrospray ionization mass spectrometry, was used to investigate the resulting flavin adduct structure. PMID- 11425555 TI - Parallel-stranded hairpins containing 8-aminopurines. Novel efficient probes for triple-helix formation. AB - We describe novel oligomers with a greater propensity to form triplexes than oligomers containing only natural bases. They consist of a polypyrimidine sequence linked head-to-head with a polypurine sequence carrying one or several 8 aminoadenine or 8-aminoguanines. The presence of 8-aminopurines also stabilised the parallel-stranded duplex structure. PMID- 11425556 TI - Incorporation of alpha- and beta-LNA (locked nucleic acid) monomers in oligodeoxynucleotides with polarity reversals. AB - The thymidine monomers of LNA with both alpha- and beta-configuration are incorporated with polarity reversals (i.e., with 3'-3' and 5'-5' junctions) in oligodeoxynucleotides with beta- and alpha-configuration, respectively. A 5'-O phosphoramidite of the beta-LNA monomer is synthesised. Large destabilisations of duplexes with both complementary DNA and RNA are observed for oligodeoxynucleotides containing the alpha-LNA monomer, whereas a duplex with complementary RNA of an alpha-oligodeoxynucleotide containing the beta-LNA monomer is not destabilised. PMID- 11425557 TI - A mechanism-based affinity-labeling agent for possible use in isolating N acetylglucosaminidase. AB - We have prepared several mechanism-based affinity-labeling agents for possible use in isolating N-acetylglucosaminidase, in which an N-acetylglucosamine is linked to an o-monofluoro- or difluoro-methyl phenoxy glycoside with or without a cleavable disulfide group in the tether to biotin. PMID- 11425558 TI - Rational design of a new series of pronucleotide. AB - A new pronucleotide series is described involving a two-step degradation process mediated by, respectively, carboxylesterase and phosphoramidase. Taking AZT as nucleosidyl moiety, it is shown that most of the compounds inhibit HIV replication in TK(-) cell line, which proves 5'-AZTMP delivery. PMID- 11425559 TI - Novel and potent tacrine-related hetero- and homobivalent ligands for acetylcholinesterase and butyrylcholinesterase. AB - Based upon synthetic and biochemical results, a novel and potent tacrine analogue and heterobivalent analogues of tacrine, were designed. The role played by the amino groups of homo- and heterobivalent ligands in the interaction with the peripheral and catalytic sites of AChE and BuChE were investigated. The syntheses of these materials together with the results of AChE/BuChE inhibition assays are detailed. PMID- 11425560 TI - Biotin labelling of amines by polymer-assisted solution-phase synthesis. AB - An efficient and simple polymer-assisted approach has been developed to biotinylate multifunctional compounds bearing an amino functionality. Biotin was immobilized on aminomethylated polystyrene via the Kenner safety catch linker, activated and subsequently transferred to the amino function of the target compounds chemoselectively, thus avoiding protecting group operations. This approach holds potential for the introduction of spacer-modified biotin derivatives. PMID- 11425561 TI - Carbonic anhydrase inhibitors: 4-sulfamoyl-benzenecarboxamides and 4-chloro-3 sulfamoyl-benzenecarboxamides with strong topical antiglaucoma properties. AB - Reaction of 4-carboxy-benzenesulfonamide or 4-chloro-3-sulfamoyl benzoic acid with carboxy-protected amino acids/dipeptides, or aromatic/heterocyclic sulfonamides/mercaptans afforded the corresponding benzene-carboxamide derivatives. These were tested as inhibitors of three carbonic anhydrase (CA) isozymes, CA I, II and IV. Some of the new derivatives showed affinity in the low nanomolar range for isozymes CA II and IV, involved in aqueous humor secretion within the eye, and were tested as topically acting anti-glaucoma agents, in normotensive and glaucomatous rabbits. Good in vivo activity and prolonged duration of action has been observed for some of these derivatives, as compared to the clinically used drugs dorzolamide and brinzolamide. Some of the 4-chloro-3 sulfamoyl benzenecarboxamides reported here showed higher affinity for CA I than for the sulfonamide avid isozyme CA II. PMID- 11425562 TI - Discovery of 2,3-diaryl-1,3-thiazolidin-4-ones as potent anti-HIV-1 agents. AB - Design, synthesis and anti-HIV activity of a series of 2,3-diaryl-1,3-thiazolidin 4-ones are reported. Some derivatives proved to be highly effective in inhibiting HIV-1 replication at nanomolar concentrations thereby acting as non-nucleoside HIV-1 RT inhibitors (NNRTIs). SAR studies evidenced that the nature of the substituents at the 2 and 3 positions of the thiazolidinone nucleus largely influenced the in vitro anti-HIV activity of this new class of potent antiviral agents. PMID- 11425564 TI - Arylisothiocyanate-containing esters of caffeic acid designed as affinity ligands for HIV-1 integrase. AB - Integrase is an enzyme found in human immunodeficiency virus, which is required for the viral life cycle, yet has no human cellular homologue. For this reason, HIV integrase (IN) has become an important target for the development of new AIDS therapeutics. Irreversible affinity ligands have proven to be valuable tools for studying a number of enzyme and protein systems, yet to date there have been no reports of such affinity ligands for the study of IN. As an initial approach toward irreversible ligand design directed against IN, we appended isothiocyanate functionality onto caffeic acid phenethyl ester (CAPE), a known HIV integrase inhibitor. The choice of isothiocyanate as the reactive functionality, was based on its demonstrated utility in the preparation of affinity ligands directed against a number of other protein targets. Several isomeric CAPE isothiocyanates were prepared to explore the enzyme topography for reactive nitrogen and sulfur nucleophiles vicinal to the enzyme-bound CAPE. The preparation of these CAPE isothiocyanates, required development of new synthetic methodology which employed phenyl thiocarbamates as latent isothiocyanates which could be unmasked near the end of the synthetic sequence. When it was observed that beta-mercaptoethanol (beta-ME), which is required to maintain the catalytic activity of soluble IN (a F185KC280S mutant), reacted with CAPE isothiocyanate functionality to form the corresponding hydroxyethylthiocarbamate, a variety of mutant IN were examined which did not require the presence of beta-ME for catalytic activity. Although in these latter enzymes, CAPE isothiocyanate functionality was presumed to be present and available for acylation by IN nucleophiles, they were equally effective against Cys to Ser mutants. One conclusion of these studies, is that upon binding of CAPE to the integrase, nitrogen or sulfur nucleophiles may not be properly situated in the vicinity of the phenethyl aryl ring to allow reaction with and covalent modification of reactive functionality, such as isothiocyanate groups. The fact that introduction of the isothiocyanate group onto various positions of the phenethyl ring or replacement of the phenyl ring with naphthyl rings, failed to significantly affect inhibitory potency, indicates a degree of insensitivity of this region of the molecule toward structural modification. These findings may be useful in future studies concerned with the development and use of HIV-1 integrase affinity ligands. PMID- 11425565 TI - Mutation of cysteine-295 to alanine in secondary alcohol dehydrogenase from Thermoanaerobacter ethanolicus affects the enantioselectivity and substrate specificity of ketone reductions. AB - The mutation of Cys-295 to alanine in Thermoanaerobacter ethanolicus secondary alcohol dehydrogenase (SADH) was performed to give C295A SADH, on the basis of molecular modeling studies utilizing the X-ray crystal structure coordinates of the highly homologous T. brockii secondary alcohol dehydrogenase (1YKF.PDB). This mutant SADH has activity for 2-propanol comparable to wild-type SADH. However, the C295A mutation was found to cause a significant shift of enantioselectivity toward the (S)-configuration in the reduction of some ethynylketones to the corresponding chiral propargyl alcohols. This result confirms our prediction that Cys-295 is part of a small alkyl group binding pocket whose size determines the binding orientation of ketone substrates, and, hence, the stereochemical configuration of the product alcohol. Furthermore, C295A SADH has much higher activity towards t-butyl and some alpha-branched ketones than does wild-type SADH. The C295A mutation does not affect the thioester reductase activity of SADH. The broader substrate specificity and altered stereoselectivity for C295A SADH make it a potentially useful tool for asymmetric reductions. PMID- 11425566 TI - Anti-HIV and cytotoxic activities of Ru(II)/Ru(III) polypyridyl complexes containing 2,6-(2'-benzimidazolyl)-pyridine/chalcone as co-ligand. AB - Ru(II)/Ru(III) polypyridyl complexes containing 2,6-(2'-benzimidazolyl)-pyridine or chalcone as co-ligands were synthesized and characterized previously (Mishra, L.; Sinha, R. Indian J. Chem., Sec. A 2001, in press. Mishra, L.; Sinha, R. Indian J. Chem., Sec. A, 39A, 2000, 1131). Their interaction with aqueous buffered calf thymus DNA was measured. (Novakova, O.; Kasparkova, J.; Vrana, O.; van Vliet, P. M., Reedijk, J.; Brabec, V., Biochem. 34, 1995, 12369 and these results prompted additional screening for anti-HIV (human immunodeficiency virus) activity against DNA replication in H9 lymphocytes and cytotoxic activity against eight tumor cell lines. The most active compounds were 17 in the former assay (EC(50) < 0.1 microg/mL and TI > 23.1) and 3, 8, 10, and 14 in the latter assay, especially selectively against the 1A9 ovarian cancer cell line (IC(50) = 4.1, 3.8, 3.6, and 2.5 microg/mL, respectively). PMID- 11425567 TI - Stereochemical selectivity of methanandamides for the CB1 and CB2 cannabinoid receptors and their metabolic stability. AB - Several chiral, analogues of the endogenous cannabinoid receptor ligand, arachidonylethanolamide (anandamide), methylated at the 2,1' and 2' positions using asymmetric synthesis were evaluated in order to study (a) stereoselectivity of binding to CB1 and CB2 cannabinoid receptors; and (b) metabolic stability with regard to anandamide amidase. Enantiomerically pure 2-methyl arachidonic acids were synthesized through diastereoselective methylation of the respective chiral 2-oxazolidinone enolate derivatives and CB1 and CB2 receptor affinities of the resulting chiral anandamides were evaluated using a standard receptor binding assay. Introduction of a single 2-methyl group increased affinity for CB1, led to limited enantioselectivity and only modestly improved metabolic stability. However, a high degree of enantio- and diastereoselectivity was observed for the 2,1'-dimethyl analogues. (R)-N-(1-methyl-2-hydroxyethyl)-2-(R)-methyl arachidonamide (4) exhibited the highest CB1 receptor affinity in this series with a K(i) of 7.42 nM, an at least 10-fold improvement on anandamide (K(i)=78.2 nM). The introduction of two methyl groups at the 2-position of anandamide led to no change in affinity for CB1 but somewhat enhanced metabolic stability. Conversely, chiral headgroup methylation in the 2-gem-dimethyl series led to chiral analogues possessing a wide range of CB1 affinities. Of these the (S) 2,2,2'-trimethyl analogue (12) had the highest affinity for CB1 almost equal to that of anandamide. In agreement with our previous anandamide structure-activity relationship work, the analogues in this study showed high selectivity for the CB1 receptor over CB2. The results are evaluated in terms of stereochemical factors affecting the ligand's affinity for CB1 using receptor-essential volume mapping as an aid. Based on the results, a partial CB1 receptor site model is proposed, that bears two hydrophobic pockets capable of accommodating 1'- and 2 methyl groups PMID- 11425568 TI - Rat liver microsomal enzyme catalyzed oxidation of 4-phenyl-trans-1-(2 phenylcyclopropyl)-1,2,3,6-tetrahydropyridine. AB - As part of our ongoing studies to characterize the catalytic pathway(s) for the monoamine oxidase and cytochrome P450 catalyzed oxidations of 1,4-disubstituted 1,2,3,6-tetrahydropyridinyl derivatives, we have examined the metabolic fate of 4 phenyl-trans-1-(2-phenylcyclopropyl)-1,2,3,6-tetrahydropyridine in NADPH supplemented rat liver microsomes. Three metabolic pathways have been identified: (1) allylic ring alpha-carbon oxidation to yield the dihydropyridinium species, (2) nitrogen oxidation to yield the N-oxide and (3) N-dealkylation to yield 4 phenyl-1,2,3,6-tetrahydropyridine and cinnamaldehyde. A possible mechanism to account for the formation of cinnamaldehye involves an initial single electron transfer from the nitrogen lone pair to the iron oxo system Fe(+3)(O) to form the corresponding cyclopropylaminyl radical cation that will be processed further to the final products. The reaction pathway leading to the dihydropyridinium metabolite may also proceed via the same radical cation intermediate but direct experimental evidence to this effect remains to be obtained. PMID- 11425569 TI - Conformationally restricted hybrid analogues of the hormone 1 alpha,25 dihydroxyvitamin D(3): design, synthesis, and biological evaluation. AB - Four new conformationally restricted hybrid analogues of the hormone 1 alpha-25 dihydroxyvitamin D(3) (1,25D3) have been synthesized in a convergent manner by combining enantiomerically pure C,D-ring ketones (-)-15 and (-)-17 with racemic 1 hydroxymethyl A-ring phosphine oxide (+/-)-18. Parent hybrid analogue 6, which combines the calcemia-inactivating 1 beta-hydroxymethyl A-ring modification with the antiproliferation- activating 20-epi-22-oxa-25-hydroxydiethyl C,D-ring side chain modification, is comparable in potency to 1,25D3 at the low nM level in inhibiting proliferation in a wide assortment of malignant cell lines in vitro with extremely low calcemic activity in vivo. Surprisingly, both conformationally restricted analogues of 6 (8b and 9b), which incorporate rigidifying units at their 25-hydroxyl side chain termini, retained the desirable antiproliferative, transcriptional, and calcemic activities of the parent compound. PMID- 11425570 TI - Chemiluminescence flow sensor for berberine with immobilized reagents. AB - A sensitive chemiluminescence (CL) sensor for berberine combined with flow injection (FI) technology is described. The analytical reagents involved in the CL reaction, including luminol and potassium ferricyanide, were both immobilized on an anion-exchange resin column. While a volume of sodium phosphate was passed through the column, the two CL reagents were eluted from the resin and then mixed with a berberine stream under alkaline conditions. By means of the fast oxidation reaction between berberine and potassium ferricyanide, potassium ferrocyanide was generated, which then inhibited the CL reaction of luminol and potassium ferricyanide. The decreased CL intensity was correlated with the berberine concentration in the range from 0.05 to 300 ng mL(-1 )with a relative standard deviation of less than 4.3%, and the limit of detection was 0.02 ng mL(-1) at a flow rate of 2.0 mL min(-1). It was shown that the flow sensor could greatly improve the selectivity and sensitivity for determination of berberine. The determination of analyte could be performed in 2 min, including sampling and washing and be stable for over 300 analyses. Some interference and additives in preparations were tested. Two herbal medicines commonly found in compound prescriptions, rutin and baicalin, were studied preliminarily. The method has been applied for the determination of berberine in pharmaceutical preparations successfully. PMID- 11425571 TI - A series of enediynes as novel inhibitors of topoisomerase I. AB - A series of acyclic enediynes, 2-((6-substituted)-3-hexen-1,5 diynyl)benzonitriles (8--11), display potent inhibition against topoisomerase I without the formation of active biradical intermediates and show inhibitory activity against topoisomerase I at 10 microM, which is five times that of camptothecin from the results of agarose gel electrophoresis. PMID- 11425572 TI - N-(3-acyloxy-2-benzylpropyl)-N'-dihydroxytetrahydrobenzazepine and tetrahydroisoquinoline thiourea analogues as vanilloid receptor ligands. AB - The vanilloid receptor represents a promising target for drug development. Building on our previous strategies which have generated potent agonists for VR1, we now describe a series of novel N-(3-acyloxy-2-benzylpropyl)-N' dihydroxytetrahydrobenzazepine and tetrahydroisoquinoline thiourea analogues, several of which are potent VR1 antagonists. We report here the rationale for the design, the synthesis, and the in vitro characterization of activity in assays for [(3)H]resiniferatoxin binding and (45)Ca influx using heterologously expressed rat VR1. PMID- 11425573 TI - Ligand recognition by the vitamin D receptor. AB - Three-dimensional structure of the ligand binding domain (LBD) of the vitamin D receptor (VDR) docked with the natural ligand 1 alpha,25-dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)] has been mostly solved by the X-ray crystallographic analysis of the deletion mutant (VDR-LBD Delta 165-215). The important focus, from now on, is how the VDR recognizes and interacts with potent synthetic ligands. We now report the docking models of the VDR with three functionally and structurally interesting ligands, 22-oxa-1,25-(OH)(2)D(3) (OCT), 20-epi-1,25-(OH)(2)D(3) and 20-epi-22-oxa-24,26,27-trihomo-1,25-(OH)(2)D(3). In parallel with the computational docking studies, we prepared twelve one-point mutants of amino acid residues lining the ligand binding pocket of the VDR and examined their transactivation potency induced by 1,25-(OH)(2)D(3) and these synthetic ligands. The results indicate that L233, R274, W286, H397 and Y401 are essential for holding the all ligands tested, S278 and Q400 are not important at all, and the importance of S237, V234, S275, C288 and H305 is variable depending on the side chain structure of the ligands. Based on these studies, we suggested key structural factors to bestow the selective action on OCT and the augmented activities on 20-epi-ligands. Furthermore, the docking models coincided well with our proposed active space-region theory of vitamin D based on the conformational analyses of ligands. PMID- 11425574 TI - 5-(Trifluoromethyl)-beta-l-2'-deoxyuridine, the L-enantiomer of trifluorothymidine: stereospecific synthesis and antiherpetic evaluations. AB - As a part of our ongoing work on beta-L-nucleoside analogues as potential antiviral drugs, we have synthesized 5-(trifluoromethyl)-beta-L-2'-deoxyuridine (L-TFT), the hitherto unknown L-enantiomer of trifluorothymidine (CF(3)dUrd, TFT). We have also studied the effect of L-TFT on human and herpes simplex virus (HSV) type 1 and 2 thymidine kinases, and human thymidine phosphorylase, as well as its anti-HSV-1 and anti-HSV-2 activities in cell cultures. L-TFT has been found: (i) to inhibit HSV-1 TK with activity comparable to TFT, with no effect on human TK, (ii) to be phosphorylated by the viral enzyme with similar efficiency to TFT, (iii) to be resistant, in contrast to TFT, to hydrolysis by human thymidine phosphorylase. Unfortunately, when evaluated in cell cultures, L-TFT did not show any anti-HSV-1 and anti-HSV-2 activities. PMID- 11425575 TI - A new aspect of view in synthesizing new type beta-adrenoceptor blockers with ancillary antioxidant activities. AB - A series of vanilloid-type beta-adrenoceptor blockers derived from antioxidant traditional Chinese herbal medicines were synthesized and tested for their antioxidant and adrenoceptor antagonistic activities. They all possessed significant beta-adrenoceptor blocking activities under in vitro experiments and radioligand binding assays. In addition, some compounds were further examined in in vivo tests and produced antagonist effects matching that of propranolol and labetalol by measurements of antagonism toward (-)isoproterenol-induced tachycardia and (-)phenylephrine-induced pressor responses in anesthetized rats. Furthermore, all of the compounds had antioxidant effects inherited from their original structures. In conclusion, compound 11 had the most potent beta adrenoceptors blocking activity, 12 and 13 possessed high cardioselectivity, whereas 14, 15 and 16 possessed additional alpha-adrenoceptor blocking activity and 15 is the most effective antioxidant of all. The antioxidant activity may be due to their alpha and beta unsaturated side chain at position 1 and ortho substituted methoxy moiety on 4-phenoxyethylamine. PMID- 11425576 TI - Ortho-carboranyl glycosides for the treatment of cancer by boron neutron capture therapy. AB - Distinct biological properties of the ortho-carboranyl (1,2-dicarba-closo dodecaboranyl) glycosides 1, 2 and 3 were evaluated to estimate the suitability of these compounds for cancer treatment by boron neutron capture therapy. The boron uptake into B16-Melanoma cells was significantly higher by incubating the cells with aqueous solutions of carboranyl glucoside 1 (11.2 ppm after 3h), lactoside 2 (13.2 ppm after 12h) and maltoside 3 (20.0 ppm after 24h) compared with solutions of clinically used p-boronophenylalanine (BPA) 5 (3.1 ppm after 24h). Carboranyl maltoside 3 was more effective than boron-10 enriched 5 in killing C-6 rat glioma cells by incubating the cells with the compound and subsequent treatment with thermal neutrons. 3 was also administrated iv, in concentrations of 25 mg boron/kg body weight to rats bearing brain tumors. After a period of 4h after administration the concentration of boron in the tumor tissue was 3.0 ppm. PMID- 11425577 TI - Molecular modeling, structure--activity relationships and functional antagonism studies of 4-hydroxy-1-methyl-4-(4-methylphenyl)-3-piperidyl 4-methylphenyl ketones as a novel class of dopamine transporter inhibitors. AB - We previously disclosed the discovery of 4-hydroxy-1-methyl-4-(4-methylphenyl)-3 piperidyl 4-methylphenyl ketone (3) as a novel class of dopamine transporter (DAT) inhibitors and showed that (+/-)-3 has a significant functional antagonism against cocaine in vitro. Our previous preliminary structure-activity relationship study led to identification of a more potent DAT inhibitor [(+/-)-4] but this compound failed to show any significant functional antagonism. To search for more potent analogues than 3 but still displaying significant functional antagonism, further SARs, molecular modeling studies and in vitro pharmacological evaluation of this novel class of DAT inhibitors were performed. Sixteen new analogues were synthesized in racemic form and evaluated as DAT inhibitors. It was found that seven new analogues are reasonably potent DAT inhibitors with K(i) values of 0.041--0.30 and 0.052--0.16 microM in [(3)H]mazindol binding and inhibition of DA reuptake. Chiral isomers of several potent DAT inhibitors were obtained through chiral HPLC separation and evaluated as inhibitors at all the three monoamine transporter sites. In general, the (-)-isomer is more active than the (+)-isomer in inhibition of DA reuptake and all the (-)-isomers are selective inhibitors at the DAT site. Evaluation of cocaine's effect on dopamine uptake in the presence and absence of (+)-3 and (-)-3 showed that (-)-3 is responsible for the functional antagonism obtained with the original lead (+/-)-3. Out of the new compounds synthesized, analogue (+/-)-20, which is 8- and 3-fold more potent than (+/-)-3 in binding and inhibition of DA reuptake, appeared to have improved functional antagonism as compared to (+/-)-3. PMID- 11425578 TI - Synthesis and biological activity of a photoaffinity etoposide probe. AB - The epipodophyllotoxin etoposide is a potent and widely used anticancer drug that targets DNA topoisomerase II. The synthesis, photochemical, and biological testing of a photoactivatable aromatic azido analogue of etoposide also containing an iodo group is described. This azido analogue should prove useful for identifying the etoposide interaction site on topoisomerase II. Irradiation of the azido analogue and an aldehyde-containing azido precursor with UV light produced changes in their UV--visible spectra that were consistent with photoactivation. The azido analogue strongly inhibited topoisomerase II and inhibited the growth of Chinese Hamster Ovary cells. Azido analogue-induced topoisomerase II--DNA covalent complexes were significantly increased subsequent to UV irradiation of drug-treated human leukemia K562 cells as compared to etoposide-treated cells. These results suggest that the photoactivated form of etoposide is a more effective topoisomerase II poison either by interacting directly with the enzyme or with DNA subsequent to topoisomerase II-mediated strand cleavage. PMID- 11425579 TI - Synthesis of novel analogues of marine indole alkaloids: mono(indolyl)-4 trifluoromethylpyridines and bis(indolyl)-4-trifluoromethylpyridines as potential anticancer agents. AB - Mono(indolyl)-4-trifluoromethylpyridines and bis(indolyl)-4 trifluoromethylpyridines were synthesized using Suzuki cross-coupling reaction between 2-chloro-4-trifluoromethylpyridine 9, 2,6-dichloro-4 trifluoromethylpyridine 6 or 2,6-dichloro-3-cyano-4-trifluoromethylpyridine 23 and N-tosyl-3-indolylboronic acid 10. They were evaluated for cytotoxic activity against P388 and A-549 cells with IC(50) values. 4-Trifluoromethyl-2,6-bis[3'-(N tosyl-6'-methoxylindolyl)]pyridine 18 was identified as the most potent in this series. PMID- 11425580 TI - Synthesis of plaunotol derivatives and their antibacterial activities against Helicobacter pylori. AB - Plaunotol, a known antiulcer drug, has antibacterial activities against Helicobacter pylori. Plaunotol thiourea derivatives 2--4 and diol derivatives 6- 10 were designed in search for a compound with high antibacterial activities. Thiourea derivatives 2--4 were synthesized regioselectively using our effective synthetic route for plaunotol (1), and diol derivatives 6--10 were also synthesized. Their antibacterial activities against H. pylori are described and we found that the most potent antibacterial agent was C1-thiourea derivative 2c. PMID- 11425581 TI - Disease activated drugs: a new concept for the treatment of asthma. AB - Disease activated drugs (DAD) are pro-drugs of one active principle or combinations of two drugs, which have a proven efficacy for the treatment of the target disease. In opposition to pro-drugs, DAD are activated in inflamed but not normal tissues. Due to the disease specific activation, the amount of locally released drug(s) should be related directly to the severity of the inflammation. To test this concept in asthma a PDE4 inhibitor, an isoquinoline derivative, was chemically derivatized into pro-drugs or combined with corticosteroids. These new compounds were more readily cleaved into active PDE4 inhibitor, in bronchoalveolar lavage fluid (BALF) from Brown-Norway rats with lung inflammation than in BALF from rats without airway inflammation. The DAD concept (local selective release and improved therapeutic window) was validated in vivo using the inhibition of methacholine induced bronchoconstriction in guinea pigs with or without ozone induced lung inflammation. An example of DAD hydrolysis (isoquinoline-dexamethasone) was also examined in BALF from asthmatics and healthy volunteers. PMID- 11425582 TI - A C-ring regioisomer of the marine alkaloid meridine exhibits selective in vitro cytotoxicity for solid tumours. AB - 9-Hydroxybenzo[b]pyrido[4,3,2-de](1,10)-phenantrolin-8-one (1), a regioisomer of the marine alkaloid meridine, was synthesized from 5,8-dimethoxy-6-nitro-4(1H) quinolinone in eight steps and 23% overall yield. A shorter route was also investigated, based on the hetero Diels-Alder reaction between o nitrocinnamaldehyde dimethylhydrazone and 4-halogen-6-bromo-5,8-quinolinequinones followed by reductive cyclization onto the C-5 carbonyl of the quinone. Compound 1 showed a remarkable in vitro cytotoxicity, with a pattern of selectivity towards solid tumours that is not found in the reference alkaloid, the activity against the human lung carcinoma (A-549) being particularly noteworthy. The activities of meridine and compound 1 as inhibitors of topoisomerase II were also significantly different. PMID- 11425583 TI - Solution chemistry and DNA binding properties of MEN 10755, a novel disaccharide analogue of doxorubicin. AB - The behavior under physiological conditions of MEN 10755, a novel disaccharide analogue of doxorubicin, was investigated in detail by a variety of spectroscopic techniques including spectrophotometry, fluorescence, and (1)H NMR. The pH dependent properties of MEN 10755 were also analysed by spectrophotometry and potentiometry within the pH range 5--11. It is found that MEN 10755 behaves very similarly to doxorubicin and reproduces closely its pH dependent pattern. Like doxorubicin, MEN 10755 undergoes dimerization with a significantly smaller association constant. The interaction of MEN 10755 with calf thymus DNA was studied in detail. Spectrophotometric and fluorescence titrations of MEN 10755 with calf thymus DNA show spectral patterns almost identical to those obtained with doxorubicin implying that the binding mechanism and the stability of the resulting adducts are very similar. An apparent affinity constant of 1.2 x 10(6) was determined for the interaction of MEN 10755 with calf thymus DNA to be compared with the value of 3.3 x 10(6) measured for doxorubicin, under the same conditions. The effects of both anthracyclines on the thermal denaturation profiles of calf thymus DNA were also analyzed; both compounds turned out to stabilize to a similar extent the DNA double helix and to give rise to a characteristic two-step melting profile. The implications of the present results for the pharmacological activity and the mechanism of action of this novel and promising antitumor compound are discussed. PMID- 11425584 TI - Synthesis and properties of bile acid phosphoramidites 5'-tethered to antisense oligodeoxynucleotides against HCV. AB - Recently, we synthesized antisense oligonucleotides (AS-ODNs) directed against the non-coding-region (NCR) and the adjacent core region of the hepatitis C virus (HCV) RNA. Backbone modifications like phosphorothioates, methyl- and benzylphosphonates were introduced three at each end of the sequence. For improvement of liver specific drug targeting and/or hepatocellular uptake efficient AS-ODNs were covalently conjugated to biomolecules such as cholesterol or bile acids. The use of base-labile alkylphosphonates afforded mild conditions for deprotection of bile acid conjugated AS-ODNs. Here, we describe a convenient synthesis of new cholic acid and taurocholic acid phosphoramidites. Derivatization to taurocholic acid was effected directly before phosphitylation reaction, which is the last step of the phosphoramidite synthesis. These building blocks were coupled to the 5'-position of AS-ODNs in the last step of solid-phase synthesis. After mild deprotection, purification and characterization the properties of these modified AS-ODNs like their lipophilicity or their ability to form stable duplices to DNA and RNA were investigated. Enhanced lipophilicity and formation of stable duplices and heteroduplices makes bile acid conjugated AS ODNs interesting as antiviral antisense therapeutics against HCV. PMID- 11425585 TI - Torsion angle based design of peptidomimetics: a dipeptidic template adopting beta-I turn (Ac-Aib-AzGly--NH(2)). AB - We have attempted to design a model dipeptide (acetyl dipeptide amide, Ac-CA1-CA2 -NH(2)) that can adopt specifically typical torsion angles of the beta-I turn (phi(i+1), psi(i+1), phi(i+2), psi(i+2)=-60 degrees, -30 degrees, -90 degrees, 0 degrees ). The key of the design is the combination of constrained amino acids that prefer to adopt the desired torsion angles. We chose Aib (aminoisobutyric acid) as the first residue of which phi and psi angles must be -60 degrees and 30 degrees, respectively. Then, we selected an azaamino acid as the second residue since previous studies have indicated that they prefer to adopt +/-90 degrees of phi angle and 0 degrees or 180 degrees of psi angle. The conformational preference of the resulting Ac-Aib-AzGly--NH(2) is investigated using ab initio methods. The conformations implying beta-I and beta-I' turns are energetically most favorable, as we expected. Thus, we synthesized the designed molecule on the solid phase considering the future generation of combinatorial libraries using an automatic peptide synthesizer. Then, NMR spectroscopy was carried out to confirm their conformational preference in solution was carried out. The results indicated that the Ac-Aib-AzGly--NH(2) adopt beta-I or beta-I' turns in solution forming an intramolecular hydrogen bonding between Ac--C(O) and terminal NH(2). We believe that such a small peptidomimetic template is highly useful for the design of drug candidates and molecular devices. PMID- 11425586 TI - Novel pyrrolo[3,2-f]quinolines: synthesis and antiproliferative activity. AB - Novel pyrrolo[3,2,f]quinoline derivatives have been synthesized and tested as antiproliferative agents. They are characterized by an angular aromatic tricyclic system, to which a methyl group can be bound at position 7, and by a methanesulfon-anisidide side chain as such, or lacking the m-methoxy substituent at position 1. The novel compounds were shown to exhibit cell growth inhibitory properties when tested against the NCI panel of cell lines, in particular those obtained from leukemias. Although the compounds are able to stimulate topoisomerase II poisoning at high concentration, the cell growth inhibition properties do not appear to rest principally on this mechanism of action. Overall, the most active proved to be compound 9, having the m-methoxy substituent typical of amsacrine, followed by the 7-methyl derivative 10 and by the unsubstituted compound 8. Comparison with previously investigated regioisomers shows modulation of activity dictated by the position and conformational freedom of side-chain groups. PMID- 11425587 TI - Synthesis of tropane and nortropane analogues with phenyl substitutions as serotonin transporter ligands. AB - The effects of structural modifications of 2 beta-carbomethoxy-3 beta-phenyl tropane analogues were evaluated on in vitro affinity to the dopamine (DAT) and serotonin (5-HTT) transporters in rat brain tissue. The introduction of a large alkyl group at the 4'-position of the phenyl ring, affording 2 beta-carbomethoxy 3 beta-(4'-alkylphenyl) tropane, diminished the affinity for the DAT whereas moderate 5-HTT affinity was obtained. The introduction of an iodine at the 3' position of the 4'-alkylphenyl, affording 2 beta-carbomethoxy-3 beta-(3'-iodo-4' alkylphenyl) tropane, and N-demethylation, affording 2 beta-carbomethoxy-3 beta (3'-iodo-4'-alkylphenyl) nortropane, improved affinity and specificity for the 5 HTT. It could be assumed from these results that the combination of these three modifications of tropane structure yielded highly selective compounds for the 5 HTT. Of the new compounds synthesized, the most selective cocaine derivative, 2 beta-carbomethoxy-3 beta-(3'-iodo-4'-isopropylphenyl) nortropane (8d) labeled with iodine-123 or carbon-11, could be a potential ligand for exploration of the 5-HT transporter by SPET or PET. PMID- 11425588 TI - Synthesis of linear-B saccharopeptides via enzymatic galactosylation of non natural glucosamide acceptors. AB - A series of D- and L-glycopyranuronic acids are coupled to glucosamines to give saccharopeptides. These 'disaccharides', in which the acetyl moiety of the natural N-acetyl-glucosamine is replaced by various sugar acids, turned out to be surprisingly good substrates for beta(1-4)-galactosyl-transferase and alpha(1-3) galactosyl-transferase. The enzymes transfer successively two galactose units from the donor UDP-galactose onto these acceptor substrates, despite the far reaching alterations, regio- and stereospecifically in the expected manner to give linear-B saccharopeptides. PMID- 11425589 TI - A new synthetic approach to 1-[(3R,4R)-1-cyclooctylmethyl-3-hydroxymethyl-4 piperidyl]-3-ethyl-1,3-dihydro-benzimidazol-2-one(J-113397), the first non peptide ORL-1 receptor antagonist.. AB - An efficient approach to 1-[(3R,4R)-1-cyclooctylmethyl-3-hydroxymethyl-4 piperidyl]-3-ethyl-1,3-dihydro-benzimidazol-2-one (J-113397) 1, the first non peptide ORL-1 receptor antagonist described in literature, is outlined. After construction of the piperidine framework through Dieckmann cyclization of the Michael adduct 8 of cyclooctylmethylamine to methyl acrylate, condensation with o phenylendiamine produced the beta-enamino ester 2, which has been conveniently used to construct the benzimidazolone substituent at C-4. Catalytic hydrogenation of intermediate 11 followed by base-promoted cis--trans isomerization of the key compound 12 led to the formation of ester 13, which was converted to the racemic title compound by LiAlH(4) reduction. The pure enantiomers were obtained by chiral preparative HPLC separation using a derivatized cellulose-based stationary phase. PMID- 11425590 TI - Hydroxy- or methoxy-substituted benzaldoximes and benzaldehyde-O-alkyloximes as tyrosinase inhibitors. AB - Several benzaldoximes, benzaldehyde-O-ethyloximes, and acetophenonoximes were synthesized and evaluated as tyrosinase inhibitors by an assay based on tyrosinase catalyzed L-DOPA oxidation. Whereas benzaldoxime itself is only a weak inhibitor, its derivatives with one or two hydroxy or methoxy moieties in para and meta positions depress tyrosinase activity. Acetophenonoximes and trisubstituted benzaldoximes show no inhibitory activity. The IC(50) of 3,4 dihydroxybenzaldehyde-O-ethyloxime (0.3 +/- 0.1 micromol L(-1)) is of the same magnitude as tropolone (0.13 +/- 0.08 micromol L(-1)), one of the best tyrosinase inhibitors known so far. PMID- 11425591 TI - Inhibitors from rhubarb on lipopolysaccharide-induced nitric oxide production in macrophages: structural requirements of stilbenes for the activity. AB - By bioassay-guided separation, three stilbenes (rhapontigenin, piceatannol, and resveratrol), two stilbene glucoside gallates (rhaponticin 2"-O-gallate and rhaponticin 6"-O-gallate), and a naphthalene glucoside (torachrysone 8-O-beta-D glucopyranoside) with inhibitory activity against nitric oxide (NO) production in lipopolysaccharide-activated macrophages were isolated (IC(50)=11--69 microM). The oxygen functions (-OH, -OCH(3)) of stilbenes at the benzene ring were essential for the activity. The glucoside moiety reduced the activity, while the alpha,beta-double bond had no effect. Furthermore, the active stilbenes (rhapontigenin, piceatannol, and resveratrol) did not inhibit inducible NO synthase activity, but they inhibited nuclear factor-kappa B activation following expression of inducible NO synthase. PMID- 11425592 TI - Synthesis and phosphodiesterase 5 inhibitory activity of new 5-phenyl-1,6-dihydro 7H-pyrazolo[4,3-d]pyrimidin-7-one derivatives containing an N-acylamido group phenyl ring. AB - New sildenafil analogues with an N-acylamido group at the 5'-position of the phenyl ring, 6a--e, were prepared from the readily available starting compound 2 in four straightforward steps. Enzyme assays demonstrated that all the target compounds 6a-e showed higher PDE5 inhibitory activities than sildenafil. It was observed that the PDE5 inhibitory activity was enhanced as the chain length of R group increased, but introduction of the branched alkyl groups such as isopropyl (6d) and cyclohexyl (6e) resulted in the drop of potency compared with 6c. In particular the N-butyrylamido derivative 6c exhibited the highest PDE5 inhibitory activity, and was about 6-fold more potent than sildenafil. However, all the compounds exhibited somewhat weak selectivity (1--3-fold) over PDE6, indicating that the compounds 6a--e have intrinsically lower selectivity than sildenafil. PMID- 11425593 TI - Potentiation of BCNU cytotoxicity by molecules targeting abasic lesions in DNA. AB - We describe the synthesis, DNA binding measurements and pharmacological properties of a series of new heterodimeric molecules, in which a 2,6 diaminopurine is linked to a 9-aminoacridine chromophore. The linking chain contains a central N,N'-disubstituted guanidine, connected to the two chromophores by polymethylenic units of variable length. PMID- 11425594 TI - Anti-tumor promoting diterpenes from the stem bark of Thuja standishii (Cupressaceae). AB - Three new labdane-type diterpenoids, labda-8(17),13-dien-15,12R-olid-19-oic acid (1), 12S-hydroxylabda-8(17),13(16),14-trien-19-oic acid (2) and 13-ethoxylabda 8(17),11,14-trien-19-oic acid (3), along with known diterpenoids, trans-communic acid (4), totarol (5), 12-methoxyabieta-8,11,13-trien-11-ol (6), and 7 alpha,8 alpha-epoxy-6 alpha-hydroxyabieta-9(11),13-dien-12-one (7) were isolated from the stem bark of Thuja standishii. The structures of 1--3 were established by spectroscopic methods and chemical conversion. These compounds together with standishinal (8), 12-hydroxy-6,7-seco-abieta-8,11,13-trien-6,7-dial (9) and 6 alpha-hydroxysugiol (10) were tested for their inhibitory effects on Epstein- Barr virus early antigen (EBV-EA) activation induced by 12-O-tetradecanoylphorbol 13-acetate (TPA), as a test for potential cancer chemopreventive agents. Compound 10 showed strong inhibitory effect on EBV-EA induction (100% inhibition at 1000 mol ratio/TPA), and compounds 2 and 6 showed moderate inhibitory effects on EBV EA induction. In addition, 15-oxolabda-8(17),11Z,13E-trien-19-oic acid (11) was found to exhibit the anti-tumor promoting activity in two-stage mouse skin carcinogenesis test using 7,12-dimethylbenz[a]anthracene and TPA. PMID- 11425595 TI - Evaluation of novel dipeptide-bound alpha,beta-unsaturated amides and epoxides as irreversible inhibitors of guinea pig liver transglutaminase. AB - Herein, we report the results of irreversible inhibition of guinea pig liver transglutaminase (TGase) by a series of 24 novel dipeptides containing either an alpha,beta-unsaturated amide or an epoxide functional group. Their inactivation rate constants were measured using a direct continuous spectrophotometric method and were found to vary between 421 x 10(3) and 3000 x 10(3)M(-1)min(-1). PMID- 11425596 TI - A strategy for tumor-selective chemotherapy by enzymatic liberation of seco duocarmycin SA-derivatives from nontoxic prodrugs. AB - Immuno-conjugates obtained by linking enzymes with appropriate monoclonal antibodies, which bind to tumor-associated antigens, can be employed in a tumor selective antibody directed enzyme prodrug therapy (ADEPT). For this strategy the glycosides 17a--c were prepared as prodrugs of CI-TMI 14 which is a structurally simplified analogue of the highly potent antitumor agent duocarmycin SA 2. Exposure of 17a--c to cultured carcinoma cells of line A549 displayed a very low toxicity; however, after addition of the corresponding enzymes and exposure for 24 h at prodrug concentrations of <0.1 microM the proliferation of the carcinoma cells was inhibited almost completely with ED(50prodrug)/ED(50drug) of up to 270 in the presence and in the absence of the enzyme. The synthesis of 17a--c was achieved by transformation of nitroanisidine 6 into 12 which was glycosidated to give 16a--c. Removal of the silyl groups, introduction of a chlorine atom and solvolysis of the acetal groups led to 17a-c, of which 17a and 17c are promising candidates for further elaboration. PMID- 11425598 TI - Cardiac myocyte nuclear size and ploidy status decrease after mechanical support. AB - Two patients with end-stage dilated cardiomyopathy of ischemic and idiopathic origin were treated with a left ventricular assist device (LVAD) as a bridge for heart transplantation. Myocardial tissue was collected during LVAD insertion and from the left ventricular apex of the explanted hearts. The myocyte diameter, nuclear area and DNA content of myocyte nuclei were measured by static cytomorphometry in tissue sections and in isolated myocytes with a digital analysis system. The presence of apoptotic nuclei was investigated by the TdT mediated X-dUTP nick end labeling technique (TUNEL). The prolonged use of a LVAD was associated with a reduction in myocyte diameter, indicating that the LVAD may induce a reversion of myocyte hypertrophy, a process described as "reverse remodeling." In addition, unloading of the heart induced a reduction in the size and DNA content of myocyte nuclei. These results suggest that the cardiomyocyte nuclei are in a dynamic state and, as it occurs with cell hypertrophy, nuclear hypertrophy and polyploidization may be a reversible phenomenon. PMID- 11425599 TI - Diagnosis of human coronary atherosclerosis by morphology-based Raman spectroscopy. AB - BACKGROUND: Recent studies have shown that chemical composition and morphology, rather than anatomy (degree of stenosis), determine atherosclerotic plaque instability and predict disease progression. Current clinical diagnostic techniques provide accurate assessment of plaque anatomy, but have limited capability to assess plaque morphology in vivo. Here we describe a technique for a morphology-based diagnosis of atherosclerosis in the coronary arteries using Raman spectroscopy that can potentially be performed in vivo using optical fiber technology. METHODS: Raman tissue spectra were collected from normal and atherosclerotic coronary artery samples in different stages of disease progression (n=165) from explanted transplant recipient hearts (n=16). Raman spectra from the elastic laminae (EL), collagen fibers (CF), smooth muscle cells (SMC), adventitial adipocytes (AA) or fat cells, foam cells (FC), necrotic core (NC), cholesterol crystals (CC), beta-carotene containing crystals (beta-C), and calcium mineralizations (CM) were used as basis spectra in a linear least squares minimization (LSM) model to calculate the contribution of these morphologic structures to the coronary artery tissue spectra. RESULTS: We developed a diagnostic algorithm that used the fit-contributions of the various morphologic structures to classify 97 coronary artery samples in an initial calibration data set as either nonatherosclerotic, calcified plaque, or noncalcified atheromatous plaque. The algorithm was subsequently tested prospectively in a second validation data set, and correctly classified 64 (94%) of 68 coronary artery samples. CONCLUSIONS: Raman spectroscopy provides information about the morphologic composition of intact human coronary artery without the need for excision and microscopic examination. In the future, it may be possible to use this technique to analyze the morphologic composition of atherosclerotic coronary artery lesions and assess plaque instability and disease progression in vivo. PMID- 11425600 TI - Raman microspectroscopy of human coronary atherosclerosis: biochemical assessment of cellular and extracellular morphologic structures in situ. AB - BACKGROUND: We have previously shown that Raman spectroscopy can be used for chemical analysis of intact human coronary artery atherosclerotic lesions ex vivo without tissue homogenization or extraction. Here, we report the chemical analysis of individual cellular and extracellular components of atherosclerotic lesions in different stages of disease progression in situ using Raman microspectroscopy. METHODS: Thirty-five coronary artery samples were taken from 16 explanted transplant recipient hearts, and thin sections were prepared. Using a high-resolution confocal Raman microspectrometer system with an 830-nm laser light, high signal-to-noise Raman spectra were obtained from the following morphologic structures: internal and external elastic lamina, collagen fibers, fat, foam cells, smooth muscle cells, necrotic core, beta-carotene, cholesterol crystals, and calcium mineralizations. Their Raman spectra were modeled by using a linear combination of basis Raman spectra from the major biochemicals present in arterial tissue, including collagen, elastin, actin, myosin, tropomyosin, cholesterol monohydrate, cholesterol linoleate, phosphatidyl choline, triolein, calcium hydroxyapatite, calcium carbonate, and beta-carotene. RESULTS: The results show that the various morphologic structures have characteristic Raman spectra, which vary little from structure to structure and from artery to artery. The biochemical model described the spectrum of each morphologic structure quite well, indicating that the most essential biochemical components were included in the model. Furthermore, the biochemical composition of each structure, indicated by the fit contributions of the biochemical basis spectra of the morphologic structure spectrum, was very consistent. CONCLUSIONS: The Raman spectra of various morphologic structures in normal and atherosclerotic coronary artery may be used as basis spectra in a linear combination model to analyze the morphologic composition of atherosclerotic coronary artery lesions. PMID- 11425601 TI - Enhanced detection of cardiac myocyte damage by polarized light microscopy. Use in a model of coxsackievirus B3-induced myocarditis. AB - Although accurate detection of cardiac muscle damage is critical in the diagnosis of acute myocarditis or acute cellular rejection in both clinical and experimental settings, the histologic evaluation is frequently uncertain without specialized stains. In a study of adult male A/J mice infected with 2x10(5) plaque-forming units of myocarditic coxsackievirus B3, cardiac muscle injury causing myofibrillar disruption was detected as a loss of muscle birefringence by polarized light microscopy. The technique was corroborated by comparison with Masson's trichrome stain and was helpful for histologic examination especially at the early preinflammatory stages of lesion development or in fringe territories of focal lesions. Polarized light microscopy is thus an available means to enhance the histologic determination of cardiac myocyte damage and has specific advantage in an absence of specialized stains. PMID- 11425602 TI - Long-term histologic features of synthetic chordal replacement for mitral valve repair: a case report. AB - Expanded polytetrafluoroethylene (e-PTFE) sutures have been used with increasing frequency to replace chordae tendineae in mitral valves prolapsing because of myxoid change. A case is reported where fibrosis and calcification of the endocardial overgrowth covering the synthetic chordae led to severe mitral regurgitation 7 years after plastic repair and required mechanical prosthetic valve implantation. PMID- 11425603 TI - The histologic estimation of coronary artery stenoses: accuracy and the effect of lumen shape. AB - The authors have recently investigated the histologic estimation of coronary artery stenoses (CAS) to determine its reproducibility and the effect of training on reproducibility. The present study extends this work, examining the accuracy, the sensitivity, and the specificity of the estimation of CAS. Further, the effect of one histologic variable (i.e., arterial lumen shape) on the evaluation of CAS is examined. As described previously, 20 randomly selected Movat-stained coronary artery cross-sections were reviewed three times, at 3-month intervals, by six clinical pathologists (CPs), six pathology residents (Res), seven anatomic pathologists (APs), and two cardiovascular pathologists (CVPs). Before the third iteration, training in CAS assessment was provided. In the present study, for comparison with observer estimates, image analysis was used to establish the actual percent CAS and determine observer accuracy. The results of this study showed, paradoxically, that greater experience did not correlate with greater accuracy: The CPs consistently had the highest accuracy scores and the CVPs consistently had the lowest. Training, however, improved the accuracy scores of all groups. Stenotic arterial cross-sections with residual lumens showing concentric or eccentric polymorphous shapes were consistently underestimated compared to image analysis, while lumens with a eccentric slitlike shape were consistently overestimated. PMID- 11425604 TI - Ruptured chordae tendineae of the posterior leaflet of the tricuspid valve as a cause of tricuspid regurgitation following blunt chest trauma. AB - A 12-year-old boy suffered a blunt chest trauma. Some hours later, a pulsatile bilateral jugular venous distension, a holosystolic murmur heard at the low parasternal border and hepatomegaly were observed. On echocardiography, ruptured chordae tendineae of the posterior leaflet of the tricuspid valve, as well as tricuspid regurgitation were detected. He remained asymptomatic during hospital stay and was discharged home in good condition. Thus, isolated ruptured chordae tendineae of the posterior leaflet of the tricuspid valve is another cause of tricuspid regurgitation following blunt chest trauma. PMID- 11425605 TI - Primary dissecting hepatic artery aneurysm in pregnancy. AB - A 32-year-old 32 weeks pregnant primigravida presented with acute abdomen and died 13 h later. She was normotensive during her antenatal period and on admission. At postmortem, a primary dissecting aneurysm of the main hepatic artery extending into its intrahepatic right branch was found. The cause of the dissection was presumably pregnancy-related. PMID- 11425606 TI - Ipsilateral sensorimotor regions and motor sequence learning. AB - Recently, Boyd and Winstein tested three groups of individuals with damage to unilateral sensorimotor areas on a version of the serial reaction time task performed with the ipsilesional hand. Only when the individuals were provided in advance with explicit knowledge of the motor sequence were they able to benefit behaviorally from the sequence. Despite aspects of the experimental procedure and the subject selection that make it difficult to draw strong conclusions, these results add to growing evidence that sensorimotor structures contribute to the formation of abstract representations that affect more than ipsilateral effectors. PMID- 11425607 TI - Defending nativism in language acquisition. PMID- 11425608 TI - 'Quoth the Ravenellipsis' PMID- 11425609 TI - How could you miss itellipsis? PMID- 11425610 TI - Attending and intending. PMID- 11425612 TI - The effect of affect. PMID- 11425611 TI - Synchronization: making sense of the data. PMID- 11425614 TI - Long-haul brain drain. PMID- 11425613 TI - Visual cortex represents invisible lines. PMID- 11425615 TI - Perception of danger - black and white? PMID- 11425616 TI - Know thine own action. PMID- 11425617 TI - Towards an evolutionary theory of language. AB - Language is a biological trait that radically changed the performance of one species and the appearance of the planet. Understanding how human language came about is one of the most interesting tasks for evolutionary biology. Here we discuss how natural selection can guide the emergence of some basic features of human language, including arbitrary signs, words, syntactic communication and grammar. We show how natural selection can lead to the duality of patterning of human language: sequences of phonemes form words; sequences of words form sentences. Finally, we present a framework for the population dynamics of grammar acquisition, which allows us to study the cultural evolution of grammar and the biological evolution of universal grammar. PMID- 11425618 TI - Mind over grammar: reasoning in aphasia and development. AB - Research on propositional reasoning (involving 'theory of mind' understanding) in adult patients with aphasia reveals that reasoning can proceed in the absence of explicit grammatical knowledge. Conversely, evidence from studies with deaf children shows that the presence of such knowledge is not sufficient to account for reasoning. These findings are in keeping with recent research on the development of naming, categorization and imitation, indicating that children's reasoning about objects and actions is guided by inferences about others' communicative intentions. We discuss the extent to which reasoning is supported by, and tied to, language in the form of conversational awareness and experience rather than grammar. PMID- 11425619 TI - Mapping dissociations in verb morphology. AB - Substantial behavioural and neuropsychological evidence has been amassed to support the dual-route model of morphological processing, which distinguishes between a rule-based system for regular items (walk-walked, call-called) and an associative system for the irregular items (go-went). Some neural-network models attempt to explain the neuropsychological and brain-mapping dissociations in terms of single-system associative processing. We show that there are problems in the accounts of homogeneous networks in the light of recent brain-mapping evidence of systematic double-dissociation. We also examine the superior capabilities of more internally differentiated connectionist models, which, under certain conditions, display systematic double-dissociations. It appears that the more differentiation models show, the more easily they account for dissociation patterns, yet without implementing symbolic computations. PMID- 11425620 TI - Graded representations in behavioral dissociations. AB - Why do people sometimes seem to know things when they are tested in one way, while seeming unaware of this information when tested in a different way? Such task-dependent behaviors, or dissociations, often occur in infants and children, and in adults following brain damage. To explain these dissociations, researchers have posited separable knowledge systems that are differentially tapped by various tasks, develop at different rates and can be selectively impaired. There is an alternative account in which knowledge is viewed as graded in nature. Certain tasks tap weaker representations, while other tasks require stronger representations, leading to dissociations in behavior. The graded representations approach addresses dissociations observed in perception, attention, memory, executive functioning and language, and has implications for the organization, development and impairment of our cognitive systems. PMID- 11425621 TI - Genes and subtypes of schizophrenia. PMID- 11425622 TI - New light on polyglutamine neurodegenerative disorders: interference with transcription. PMID- 11425623 TI - HIV latency in naive T cells. PMID- 11425624 TI - NF1 mice: smaller brains but no tumours. PMID- 11425627 TI - Proliferation or survival: T-cell careers dissected at a molecular level. PMID- 11425636 TI - Molecular 'palpation' of BPH: a tale of MAPK signalling? AB - Benign prostatic hyperplasia (BPH) is a very common cause of hospitalization and surgery is currently the most effective therapy. MAP kinases (MAPKs) are a group of protein kinases with an important function in integrating physiological and pathological stimuli that might impact on cellular growth, differentiation and programmed cell death (apoptosis). Certain components of the MAPK signal transduction pathways are involved in stimulus-specific fine-tuning of the activities mediated by the various MAPK families. As homeostasis is impaired in the hyperplastic prostate, aberrant coordination of the MAPK cascades might be implicated in a proliferative-apoptotic imbalance. Here, we hypothesize that the pathogenesis of BPH might be facilitated by functional anomalies in the MAPK circuitry and postulate that pharmacological 'rewiring' of MAPK pathways offers a potentially exciting new avenue for improved therapeutic control of clinical BPH. PMID- 11425637 TI - Towards a molecular understanding of hair loss and its treatment. AB - Most common forms of hair loss (alopecia) are caused by aberrant hair follicle cycling and changes in hair follicle morphology. However, current treatments for alopecia do not specifically target these processes. We are now beginning to identify the molecules and molecular pathways that control normal hair follicle formation, cycling and growth. In parallel, new techniques are being developed for delivering molecules to hair follicles. Here, we outline the characteristics of common hair loss diseases, and discuss ways in which recent advances in hair follicle biology could be translated into effective therapies for these conditions. PMID- 11425638 TI - Gene therapy strategies for urological dysfunction. AB - Novel molecular techniques such as conventional and ex vivo gene therapy, and tissue engineering have only recently been introduced to the field of urology. The lower urinary tract is ideally suited for minimally invasive therapy, and also ex vivo approaches would limit the risk of systemic side effects. Muscle derived stem cells have been used successfully to treat stress incontinence, and rats with diabetic bladder dysfunction benefited from nerve growth factor (NGF) based gene therapy. Nitric oxide synthase and capase-7 might provide suitable gene therapy targets for erectile dysfunction and benign prostatic hyperplasia, respectively. PMID- 11425639 TI - Vaccines for colorectal cancer. AB - Despite recent advances in the treatment of colorectal cancer, the overall survival rate for those patients with advanced locoregional disease remains less than 50%. Although adjuvant systemic chemotherapy has improved survival of these patients, more effective therapies are needed. Immunotherapy is an approach that could have a particular role in the adjuvant therapy of colorectal cancer. There is now convincing evidence that the immune system can specifically recognize and destroy malignant cells. Although both antibody- and T-cell-mediated anti-tumor responses have been documented, the cellular immune response with its direct cytotoxic mechanisms is felt to be the principal anti-tumor arm of the immune system. Analysis of the T cells that recognize tumors has led to the identification and characterization of many tumor-associated antigens including several colorectal antigens. Current approaches to developing a vaccine for colorectal cancer use our expanded understanding of these tumor-associated antigens and the conditions that allow development of an effective cellular immune response to them. PMID- 11425640 TI - Apoptosis-regulating proteins as targets for drug discovery. AB - Defects in the regulation of apoptosis (programmed cell death) contribute to many diseases, including pathologies associated with cell loss (e.g. stroke, heart failure, neurodegeneration and AIDS), and disorders characterized by a failure to eliminate harmful cells (e.g. cancer, autoimmunity). Apoptosis is caused by activation of intracellular proteases, known as caspases, which are responsible directly or indirectly for the morphological and biochemical events that characterize the apoptotic cell. Numerous caspase regulators have been discovered, which respond to environmental stimuli and influence the decision of cell death and survival. Knowledge of the molecular details of apoptosis regulation, and the three-dimensional structures of proteins constituting the apoptosis core machinery has revealed new strategies for identifying small molecule drugs that could one day yield more effective treatments for a wide variety of illnesses. PMID- 11425641 TI - Disease model: hyperinsulinemia and insulin resistance. Part A-targeted disruption of insulin signaling or glucose transport. AB - Insulin resistance and hyperinsulinemia are common pathophysiological features of several metabolic diseases, obesity and diabetes being two notable examples. In this article we review how the use of animal models has increased our understanding of insulin resistance and hyperinsulinemia, with a particular emphasis on the use of mice with targeted disruptions of the insulin signaling pathway. PMID- 11425642 TI - Genetic testing and insurance: opportunities and challenges for society. AB - The increased availability of genetic tests poses new challenges to society. Here, we address the wider implications of genetic testing, with an emphasis on the markets for insurance. It also considers issues such as confidentiality, patient autonomy and fear of discrimination and the doctor-patient relationship. PMID- 11425643 TI - Bone marker nomenclature. PMID- 11425644 TI - Transforming growth factor-beta1 (TGF-beta) stimulates the osteoclast-forming potential of peripheral blood hematopoietic precursors in a lymphocyte-rich microenvironment. AB - Osteoclasts are generated from peripheral blood mononuclear cells (PBMNCs) in the presence of soluble receptor activator of NFkappaB ligand (sRANKL) and macrophage colony-stimulating factor (M-CSF). We show that human osteoclast formation is enhanced when PBMNCs are cultured in the presence of transforming growth factor (TGF)-beta and M-CSF prior to the addition of sRANKL. The effect was only observed in the presence of a nonadherent lymphocyte PBMNC fraction. Osteoclast formation was enhanced to a level equivalent to that induced by TGF-beta when nonadherent PBMNC fraction was removed from the cultures, prior to RANKL treatment. These data suggest that TGF-beta enhances osteoclast formation by abrogating the suppressive effect of the nonadherent PBMNCs, thereby maintaining the osteoclast-forming potential of the osteoclast precursor population. TGF-beta was without effect on proliferation of the adherent PBMNCs and did not stimulate osteoclast size or modify their immunophenotype. The effect was not mediated through prostaglandin synthesis. These results indicate that the microenvironment encountered by the osteoclast precursor prior to RANKL exposure contributes significantly to the regulation of osteoclast formation. Furthermore, the data emphasize that the effect of TGF-beta is determined by the cytokine milieu of the microenvironment and/or the state of activation of the cell being targeted by TGF beta; thus, the effect of TGF-beta is context-dependent. PMID- 11425645 TI - Transforming growth factor-beta administration modifies cyclosporine A-induced bone loss. AB - Cyclosporine A (CsA), a potent immunosuppressant used in transplantation, induces increased formation with excess resorption in the rat with resultant osteopenia. These findings are confirmed in the human model. Transforming growth factor-beta (TGF-beta) is reported to be involved in the coupling of bone formation with resorption and in vivo and in vitro stimulates osteoblasts, and in vitro inhibits osteoclasts. CsA stimulates secretion of TGF-beta1 in humans, which, while improving immunosuppression, may also contribute to renal toxicity. This study was performed determine whether exogenously administered TGF-beta would modify the bone effects of CsA. Male Sprague-Dawley rats, 6 months of age, were randomized to receive: TGF-beta and CsA vehicle (group A); TGF-beta 5 microg/kg three times per week and CsA vehicle (group B); TGF-beta vehicle and CsA 10 mg/kg (group C); or TGF-beta 5 microg/kg three times per week and CsA 10 mg/kg (group D). These were compared with control over 28 days. CsA, but not TGF-beta, increased serum 1,25(OH)(2)D levels throughout the study. CsA increased osteocalcin (BGP), but TGF-beta negated this effect. Histomorphometry confirmed the known effects of CsA, whereas TGF-beta alone had no effect. However, in combination, TGF-beta blocked CsA's effect and increased osteoblast recruitment and activity, as reflected by increased percent mineralizing surface, percent osteoid perimeter, bone formation rate (bone volume referent), and activation frequency. Thus, it appears as if TGF-beta administration may have potential in modulating the deleterious bone effects of CsA. PMID- 11425646 TI - Stimulation of sodium-dependent inorganic phosphate transport by activation of Gi/o-protein-coupled receptors by epinephrine in MC3T3-E1 osteoblast-like cells. AB - Recent data have shown that activation of Gi-protein-coupled receptors in osteoblast-like cells enhances the proliferation and differentiation of these cells. In the present study, we investigated the effect of epinephrine, an agonist of Gi-protein-coupled receptors in MC3T3-E1 cells, on Pi transport, type III Pi transporter expression, and the signaling mechanism(s) involved in this response. Epinephrine time- and dose-dependently stimulated sodium-dependent Pi transport and this effect was dependent on RNA and protein synthesis. This effect was associated with a related time-dependent increase in Pit-1 mRNA expression. Kinetic analysis indicated that the change in Pi transport activity induced by epinephrine was due to alteration in the maximal rate of Pi transport. Investigation of Pi transport stimulation by several adrenergic agonists and its inhibition by spiperone suggest that the effect of epinephrine on Pi transport was mediated by alpha1-adrenergic receptors. Pertussis toxin, which inactivates Gi/o proteins, significantly blunted the stimulatory effect of epinephrine on Pi transport. Analysis of the signaling pathways involved in this response has suggested a major role of protein kinase C and a small contribution from the mitogen-activated protein kinase Erk (MAPK(erk)). The results show that, in MC3T3 E1 osteoblast-like cells, activation of Gi/o-protein-coupled receptors induces stimulation of Pi transport. This effect is mediated by activation of protein kinase C and the MAPK(erk) pathway and probably involves the synthesis of Pit-1 transporters. PMID- 11425647 TI - Endothelin- and sarafotoxin-induced receptor-mediated calcium mobilization in a clonal murine osteoblast-like cell line, MC3T3-E1/B. AB - Previous studies have demonstrated that, in osteoblast-like MC3T3-E1 cells, various endothelin peptides and their homologous sarafotoxins generate prostaglandin E(2) (PGE(2)) release through an ET(A) receptor subtype. In this study, biphasic Ca(2+) signals elicited with endothelin (ET)-1, ET-2, ET-3, beta ET, S6a1, and S6b (ET/S6) were measured by microspectrofluorimetric methods in cell suspensions loaded with Fura-2 acetoxymethylester (Fura-2 AM). Phospholipase C (PLC)-dependent calcium activation mechanisms seem to be involved. We found evidence of Ca(2+) release from thapsigargin-sensitive and non-thapsigargin sensitive intracellular Ca(2+) stores as well as Ca(2+) transmembrane inflow through multiple voltage-independent and Ni(2+)-sensitive cation channels. Using an ET(A) receptor antagonist, BQ-123, we showed that this receptor was coupled to Ca(2+) mobilization. All agonists tested, except S6c (an ET(B)-receptor-specific agonist) induced receptor desensitization. Our results demonstrate that the ET/S6 induced Ca(2+) signaling pathway is mediated via an ET(A)-receptor subtype in MC3T3-E1/B cells. PMID- 11425648 TI - Nicotinic regulation of c-fos and osteopontin expression in human-derived osteoblast-like cells and human trabecular bone organ culture. AB - Long-term in vivo studies have highlighted smoking as a risk factor in postmenopausal osteoporosis, bone fracture incidence, and increased nonunion rates. In contrast, there are few data postulating the effects of smoking at the cellular level in human skeletal tissue. In this study, we present novel evidence demonstrating that the nicotinic receptor alpha4 subunit is present in human primary bone cells by using reverse transcriptase-polymerase chain reaction (RT PCR). In addition, we demonstrate direct cellular effects of nicotine on primary human bone cells and blockage of these effects with a nicotinic receptor antagonist, D-tubocurarine. Nicotine effects on cell proliferation were biphasic with toxic, antiproliferative effects at high levels of nicotine (>1 mmol/L) and stimulatory effects at very low levels (0.01-10 micromol/L) after 72 h. This nicotine-induced increase in cell proliferation was inhibited in a dose-dependent manner by the addition of D-tubocurarine. In addition, proliferation effects from low-level treatment correlated with an upregulation of expression of the AP-1 transcription factor, c-fos, within 1 h, which was blocked by incubation with D tubocurarine. To determine in situ bone cell responses within their trabecular matrix, cores of human bone isolated from biopsies were perfused with 0.1 micromol/L nicotine for 24 h. Western analysis of proteins isolated from the cores highlighted an increase in osteopontin, a bone matrix protein implicated in regulating resorption, which was partially inhibited by the addition of D tubocurarine. To conclude, our results suggest that nicotine has a direct effect on human bone cells in modulating proliferation, upregulation of the c-fos transcription factor, and the synthesis of the bone matrix protein, osteopontin. PMID- 11425649 TI - Bisphosphonate acts on osteoclasts independent of ruffled borders in osteosclerotic (oc/oc) mice. AB - We examined the effects of a third generation bisphosphonate [YM-175; disodium dihydrogen (cycloheptylamino)-methylene-1,1-bisphosphonate] on osteoclasts in osteosclerotic (oc/oc) mice to elucidate the cellular mechanism for incorporation of the bisphosphonate. Osteoclasts of oc/oc mice were in direct contact with bone matrix but devoid of ruffled borders. Tartrate-resistant acid phosphatase (TRAPase) showed spotty localization intercellularly, whereas vacuolar H(+) ATPase (V-ATPase) immunoreactivity was observed homogeneously in the cytoplasm. Upon injection of bisphosphonate, most osteoclasts lost cell polarity and were detached from bone surfaces. The detached osteoclasts underwent apoptosis as characterized by condensation of chromatin, absence of Golgi apparatus, and formation of many vesicles in the cytoplasm. Both TRAPase and V-ATPase were evenly distributed in the cytoplasm. The pyknotic nuclei of osteoclasts revealed DNA fragments as evidenced by the terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end-labeling (TUNEL) method. The results indicate that osteoclasts lacking ruffled borders in oc/oc mice incorporated the bisphosphonate from a site different from ruffled borders and that bisphosphonate may directly affect osteoclasts without mediating its deposition to the bone matrix. PMID- 11425650 TI - Thermostable (SULT1A1) and thermolabile (SULT1A3) phenol sulfotransferases in human osteosarcoma and osteoblast cells. AB - Sulfate conjugation is an important pathway in the metabolism of many drugs, xenobiotic compounds, and hormones. Sulfotransferases (SULTs) catalyze these reactions and have been detected and characterized in various human tissues including the liver and small intestine. Substrates for SULTs that include estrogen and thyroid hormones have well-established roles affecting skeletal integrity and disease processes. We performed the following studies to determine the presence of SULTs in human osteoblast-like cells, and to compare their characteristics to SULTs expressed in other human tissues. Four osteosarcoma cell lines (SaOS-2, U2-OS, PR, and HOS-TE85) were screened for the presence of four different SULT activities. Predominant activities were found for SULT1A1 in SaOS 2 cells, and SULT-1A3 in HOS-TE85 cells. Several biochemical properties of each enzyme that included apparent K(m) values, thermal stabilities, and responses to the inhibitors 2,6-dichloro-4-nitrophenol and NaCl were used to further characterize the SULT activities. High-performance liquid chromatography (HPLC) of the reaction products confirmed the known products of SULT1A1 and SULT1A3. When the mature human osteoblast HOB-03-CE6 cell line was tested for activity alone, the predominant activity was SULT1A3, with minimal SULT1A1. The results indicate that SULT1A1 and SULT1A3 are present in human osteosarcoma and mature osteoblast cell lines, and that the characteristics of the osteosarcoma cell SULTs are similar to those expressed in other human tissues. SULTs may have regulatory roles in the deactivation of thyroid hormones or estrogenic compounds in bone, and thus may affect hormone action and bone responses in the human skeleton. PMID- 11425651 TI - Effect of trabecular bone contour on ultimate strength of lumbar vertebra after bilateral ovariectomy in rats. AB - To test the hypothesis that the effect of trabecular microarchitecture on bone strength varies with the duration of estrogen loss, we evaluated the relationship between three-dimensional (3D) parameters for trabecular microarchitecture and bone minerals with the compressive load of the lumbar vertebra in rats. Female Sprague-Dawley rats (n = 190) were divided into 19 groups. Ten rats were killed at day 0. Half of the remaining rats underwent bilateral ovariectomy (ovx), and the others were subjected to sham surgery. Ten rats from each group were killed at 3, 7, 11, 14, 28, 42, 56, 70, and 84 days postsurgery. Urinary deoxypyridinoline and serum osteocalcin increased significantly in the ovx group from days 28 and 11, respectively, compared with the sham group. Bone mineral content (BMC) and bone mineral density (BMD) of the fifth lumbar body diminished from days 42 and 84, respectively, compared with the sham group. In ovx rats, trabecular bone volume (BV/TV), measured using 3D images of microcomputed tomography, diminished from day 28 compared with both baseline control and sham. The trabecular bone pattern factor (TBPf) and structure model index (SMI) increased from day 28 in the ovx group compared with both baseline control and sham. Ultimate compression loads diminished at day 28 compared with baseline control and decreased progressively thereafter. Neither of these parameters changed in the sham group during the same period. Within 4 weeks post-ovx, TBPf, SMI, and BV/TV correlated with load (p < 0.01). BMC and BMD correlated with load from 6 weeks post-ovx (p < 0.01). Stepwise regression analysis showed that TBPf was the most significant determinant of load within 4 weeks post-ovx (coefficient of determination [R(2)] = 0.669; p < 0.01). SMI correlated with TBPf (R(2) = 0.968; p < 0.01). Moreover, R(2) for ultimate load indicated higher values of 0.975 with TBPf and SMI. However, BMC was the most significant determinant of load from 6 weeks post-ovx (R(2) = 0.511; p < 0.01), as it was in the sham group. These data suggest that changes in trabecular bone contour with increased bone turnover are critical for reducing lumbar bone strength during the early post-ovx period in rats. PMID- 11425652 TI - Effects of ovariectomy and estrogen replacement therapy on arthritis and bone mineral density in rats with collagen-induced arthritis. AB - We investigated the effects of ovariectomy (ovx) and estrogen replacement therapy (ERT) on bone mineral density (BMD) and arthritis severity in rats with collagen induced arthritis (CIA). Seven-month-old female Sprague-Dawley rats were separated into a sham group (n = 8), CIA group (n = 14), ovx group (n = 10), CIA + ovx group (n = 11), and CIA + ovx + ERT group (n = 14). In these groups, ovx was performed at 7 days, and ERT (17beta-estradiol at 20 microg/kg three times per week) was initiated 8 days after sensitization. Every 2 weeks, until 8 weeks after sensitization, arthritis score and hind paw thickness were evaluated, and BMD of the trabecular and cortical bones in the metaphysis and diaphysis of the tibia were measured by peripheral quantitative computed tomography. The arthritis score was highest in the CIA + ovx group at all timepoints after sensitization. The hind paw thickness was significantly higher in the CIA + ovx group than in the CIA group at 8 weeks after sensitization (p < 0.05). Both the arthritis score and hind paw thickness were lower in the CIA + ovx + ERT group than in the CIA + ovx group. BMD in the metaphysis was significantly decreased in both the trabecular and cortical bones in the CIA + ovx group compared with those in the CIA group at 4, 6, and 8 weeks after sensitization. In the CIA + ovx group, trabecular BMD was changed by -34 +/- 11%, and cortical BMD changed by -14 +/- 7% in the metaphysis at 8 weeks compared with those at 0 week. In the CIA group, changes of BMD in the metaphysis were -7 +/- 11% in trabecular bone and 0 +/- 7% in cortical bone. These differences of trabecular and cortical bone loss in the metaphysis were significant (both p < 0.01). BMD reduction was significantly less in the CIA + ovx + ERT group than in the CIA + ovx group at 6 and 8 weeks after sensitization. Although BMD in the diaphysis was also reduced in the groups with CIA, the degree of reduction was smaller than in the metaphysis. We conclude that ovx in CIA rats could enhance the severity of arthritis and bone loss, and that ERT could suppress arthritis and bone loss. PMID- 11425653 TI - Osteonecrosis induced by a single administration of low-dose lipopolysaccharide in rabbits. AB - We succeeded in developing a novel rabbit model of nonsteroid and nontraumatic osteonecrosis (ON) by use of a single- and low-dose lipopolysaccharide (LPS) injection. This model is simple and highly reproducible for the frequent development of multifocal and widespread ON lesions. Male adult Japanese white rabbits intravenously injected with a single injection of 10 microg/kg body weight of LPS were histopathologically examined in the early phase (3 [n = 3], 5 [n = 3], and 24 h [n = 3]) and at 4 weeks (n = 22). Seventy-seven percent of the rabbits developed multifocal ON 4 weeks after LPS injection. ON was also observed in the femoral and humeral condyle. The average percentage of necrotic area/total area examined was 86.7 +/- 29.1% and 78.8 +/- 16.7% in the proximal one third of both the femoral and humeral bones, respectively. Organized thrombi in the intraosseous small-sized arteries and arterioles were frequently seen in and around the necrotic tissues. In the early phase, LPS treatment prominently induced thrombocytopenia, hyperlipidemia, and increased plasma levels of plasminogen activator inhibitor-1 (PAI-1). The plasma level of PAI-1 was significantly higher in the rabbits with ON than in those without ON (p < 0.01). The immunohistochemical expression of tissue factor was exaggerated in monocytes/macrophages and adipocytes in both the femoral and humeral bones of the LPS-treated rabbits. Histologically, marrow necrosis and fibrin thrombi could be observed at 24 h. In addition, pretreatment with an anticoagulant, warfarin potassium, significantly decreased the incidence of LPS-induced ON (33%, n = 9, p < 0.05) associated with elongation of prothrombin time. The results of our study show that a single administration of low-dose lipopolysaccharide induces multifocal and widespread ON characterized by the pathophysiological participation of hypercoagulability in ON development. Therefore, this model would be useful for elucidating the pathogenesis of nonsteroid ON in humans especially inflammatory hypercoagulability-induced as well as for developing preventive and therapeutic strategies. PMID- 11425654 TI - Increase in bone volume fraction precedes architectural adaptation in growing bone. AB - In mature trabecular bone, both density and trabecular orientation are adapted to external mechanical loads. Few quantitative data are available on the development of architecture and mechanical adaptation in juvenile trabecular bone. We studied the hypothesis that a time lag occurs between the adaptation of trabecular density and the adaptation of trabecular architecture during development. To investigate this hypothesis we used ten female pigs at 6, 23, 56, 104, and 230 weeks of age. Three-dimensional morphological and mechanical parameters of trabecular bone samples from the vertebra and proximal tibia were studied using microcomputed tomography and micro-finite element analysis. Both bone volume fraction and stiffness increased rapidly in the initial growth phase (from 6 weeks on), whereas the morphological anisotropy started increasing only after 23 weeks of age. In addition, the anisotropy reached its highest value much later in the development than did bone volume fraction. Hence, the alignment of trabeculae was still progressing at the time of peak bone mass. Therefore, our hypothesis was supported by the time lag between the increase in trabecular density and the adaptation of the trabecular architecture. The rapid increase of bone volume fraction in the initial growth phase can be explained by the enormous weight increase of the pigs. The trabeculae aligned at later stages when the increase in weight, and thus the loading, was slowed considerably compared with the early growth stage. Hence, the trabecular architecture was more efficient in later years. We conclude that density is adapted to external load from the early phase of growth, whereas the trabecular architecture is adapted later in the development. PMID- 11425655 TI - Changes in muscle mass, fat mass, and bone mineral content in the legs after stroke: a 1 year prospective study. AB - Demineralization and muscle atrophy, common among patients with hemiplegia, may be risk factors for future hip fracture. The aim of this longitudinal study was to investigate changes in lean (muscle) mass and bone mineral content (BMC) of the legs during the first year after stroke according to the patient's ambulatory level. Twenty-five patients immobilized due to acute stroke were followed. BMC and lean mass of each leg were measured at a mean of 7 days, 2 months, 7 months, and 1 year after the stroke using dual-energy X-ray absorptiometry. Both BMC and lean mass had decreased significantly in the paretic leg (p < 0.05) at the 1 year evaluation and the loss was significantly greater on the paretic side compared with the nonparetic side (p < 0.001). Patients who had not relearned to walk at the 2 month evaluation (n = 12) lost 6% (p < 0.05) of their lean mass in the paretic leg during this time period, and this mass was not regained within the subsequent 10 months. In contrast, a significant 5% loss of lean mass found at 2 months on the nonparetic side was regained completely. With respect to the patients who relearned to walk within the first 2 months (n = 13) lean mass had increased by 5% after 1 year (p < 0.05) in the nonparetic leg, whereas no significant changes were found in the paretic leg during follow-up. Both groups of patients did, however, lose bone mineral in the paretic leg during the first year after stroke (9% and 6%, respectively, p < 0.05), but only the patients who were still unable to walk by 2 months had significant bone loss in the nonparetic leg also (3%, p < 0.05). Thus, lean muscle mass is rapidly lost and may be regained shortly after stroke, whereas loss of BMC appears difficult to prevent, especially on the paretic side. Regaining muscle mass may, however, slow the loss of bone mineral. PMID- 11425657 TI - Nonsurgical management of subperiosteal abscess of the orbit. PMID- 11425658 TI - Low thiamine intake and risk of cataract. PMID- 11425660 TI - Ocular injuries caused by intraocular or retrobulbar foreign bodies. PMID- 11425663 TI - Ocular injuries caused by intraocular or retrobulbar foreign bodies. PMID- 11425664 TI - Optic neuropathy in Lhon and Leigh syndrome. PMID- 11425666 TI - Glaucoma in southern India. PMID- 11425667 TI - Ultrasound biomicroscopy of sclerotomy sites. PMID- 11425670 TI - Shaken not stirred. PMID- 11425671 TI - Adjuvant 5-fluorouracil and heparin prevents proliferative vitreoretinopathy : Results from a randomized, double-blind, controlled clinical trial. AB - PURPOSE: To assess the safety and efficacy of adjuvant combination therapy using 5-fluorouracil (5-FU) and low molecular weight heparin (LMWH) for prevention of proliferative vitreoretinopathy (PVR) after vitrectomy and retinal reattachment surgery. DESIGN: Prospective randomized, double-masked, placebo controlled trial. PARTICIPANTS: One hundred seventy-four high-risk patients were randomized to receive either 5-FU and LMWH therapy or placebo. Patients were selected from all patients undergoing primary vitrectomy for rhegmatogenous retinal detachment. METHOD: Results of standard surgery with 5-FU and LMWH therapy or placebo were compared at the 6-month follow-up. MAIN OUTCOME MEASURES: Development of postoperative PVR, retinal reattachment at 6 months after surgery, single operation reattachment rate, number of reoperations, and best-corrected visual acuity. RESULTS: There were 87 patients in the 5-FU and LMWH therapy group and 87 in the placebo group. The incidence of postoperative PVR was significantly lower (P = 0.02) in the 5-FU and LMWH therapy compared with the placebo group. In 26.4% (23/87) of the placebo group and in 12.6% (11/87) of the 5-FU and LMWH group, postoperative PVR developed. In the 5-FU and LMWH group, the number of patients undergoing more than one operation was 19.5% (17/87) and the number of reoperations resulting from PVR was 52.9% (9/17). In the placebo group, the number of patients undergoing more than one operation was 25.3% (22/87) and the number of reoperations resulting from PVR was 72.7% (16/22). The difference in visual acuity was not statistically different in the two treatment groups, although those patients in whom postoperative PVR developed tended to have poorer vision (P < 0.0001). There were no differences in complication rates between the two groups. CONCLUSIONS: There is a significant reduction in the incidence of postoperative PVR in patients receiving the 5-FU and LMWH therapy and in the reoperation rate resulting from PVR. This trial shows that incidence of PVR can be reduced with inexpensive and simple pharmacologic treatment with 5-FU and LMWH and should be used routinely in the treatment of patients at risk of developing PVR. PMID- 11425672 TI - How to predict proliferative vitreoretinopathy: a prospective study. AB - PURPOSE: To determine prospectively the accuracy of a predictive risk formula for the development of postoperative proliferative vitreoretinopathy (PVR) when applied in a clinical setting. DESIGN: Prospective noncomparative interventional case series. PARTICIPANTS: Two hundred nineteen subjects undergoing primary vitrectomy for rhegmatogenous retinal detachment were studied. METHOD: By use of a formula-based discriminant rule, subjects were classified as either high or low risk for the development of PVR. All subjects were followed prospectively. OUTCOME MEASURES: Development of postoperative PVR as defined by the updated the Retina Society Classification. RESULTS: Complete data were available on 212 of 219 subjects. There were 130 subjects identified as low risk and 82 subjects as high risk; 9.2% of the low-risk (12 of 130) compared with 28% (23 of 82) of the high-risk subjects had postoperative PVR develop. This difference was statistically significant (P < 0.001). CONCLUSIONS: Our study has shown that using a clinical model it is possible to identify subjects at greater risk of PVR developing after primary vitrectomy. PMID- 11425673 TI - Indocyanine green-assisted peeling of the retinal internal limiting membrane during vitrectomy surgery for macular hole repair. AB - OBJECTIVE: To determine the efficacy and safety of indocyanine green (ICG) assisted retinal internal limiting membrane (ILM) peeling during macular hole repair. DESIGN: Interventional, noncomparative, prospective case series. PARTICIPANTS: Twenty-four consecutive patients (24 eyes) with stage 3 or 4 macular holes. INTERVENTION: All eyes underwent a pars plana vitrectomy, including peeling of the posterior cortical hyaloid when necessary. Indocyanine green dye (0.5%) was instilled into the posterior vitreous cavity over the macula and left in place for 3 to 5 minutes. After removal of the ICG, the retinal ILM was peeled. Medium- to long-acting gas tamponade was used in all cases, and all patients were asked to position face down for 1 to 2 weeks. MAIN OUTCOME MEASURES: Intraoperative staining properties of ICG, technical ease of peeling of the retinal ILM, postoperative anatomic results, visual acuity, and complications were recorded. RESULTS: Indocyanine green stained the retinal ILM, but did not stain the underlying retina. Indocyanine green staining greatly facilitated the surgeons' ability to visualize and peel the ILM in each case. Peeled tissue was sent for both light and electron microscopic studies, which confirmed that the ICG-stained tissue was truly retinal ILM. Patients were observed after surgery for an average of 123 days (range, 23-195 days). Anatomic closure of the macular hole was achieved in 21 eyes (88%) with a single surgery. Visual acuity improved in 23 of 24 patients (96%) after surgery. There were no intra- or postoperative complications related to ICG use, and there was no clinical or fluorescein angiographic evidence of ICG toxicity. CONCLUSIONS: Indocyanine green stains the retinal ILM. This property facilitates ILM peeling by providing a stark contrast between the stained ILM and the unstained retina. Indocyanine green staining of the ILM appears to be a safe and useful adjunct in vitreous surgery for macular hole repair. PMID- 11425674 TI - Temperature effect on nausea during fluorescein angiography. AB - PURPOSE: Nausea is the most common adverse effect of fluorescein angiography (FA). Warming of fluorescein dye before injection has been reported anecdotally to decrease nausea frequency. We examined the effect of warmed fluorescein dye on the frequency of adverse effects of FA. DESIGN: Prospective, randomized, controlled trial. PARTICIPANTS: Five hundred forty-seven angiograms performed on 394 patients; 273 were in the control group and 274 were in the study group. METHODS: Patients were alternately assigned to FA using dye at room temperature (control group) or dye prewarmed to body temperature (study group). All patients were masked to the dye temperature. After FA, patients were questioned regarding any adverse effects experienced. MAIN OUTCOME MEASURES: Presence or absence of reported nausea associated with FA. RESULTS: Adverse reactions were reported for 39 of the angiograms (7.1%), including 32 procedures for which nausea was reported (5.9%). No significant difference in the frequency of adverse reactions was found between the two groups (P = 0.41). No significant difference in the frequency of nausea was found between the two groups (P = 0.28). The sample sizes used allow an 80% power for detecting a clinically significant difference of 4% adverse effects in one group and 10% in the other group when testing at a significance level of 5%. CONCLUSIONS: Warming of fluorescein dye does not significantly alter the incidence of nausea reported with FA. PMID- 11425675 TI - Delays in rod-mediated dark adaptation in early age-related maculopathy. AB - OBJECTIVE: To determine whether there are disturbances in the rod-mediated kinetics of dark adaptation in early age-related maculopathy (ARM). DESIGN: Comparative, observational case series. PARTICIPANTS: Twenty older adults with early ARM as defined by one or more large (> 63 microm) drusen, focal hyperpigmentation, or both, but no choroidal neovascularization or geographic atrophy, and 16 adults in the same age range with none of these fundus features. All participants had 20/25 visual acuity or better in the tested eye. METHODS: Dark adaptation functions were measured using a modified Humphrey Field Analyzer (Zeiss Humphrey Systems, Dublin, CA) to assess the rate of rod-mediated sensitivity recovery at 12 degrees on the vertical meridian in the inferior visual field after exposure to the equivalent of a 98% bleach. Baseline (prebleach) scotopic sensitivity, visual acuity, contrast sensitivity, and photopic sensitivity were also measured. MAIN OUTCOME MEASURES: Rod-cone break; second and third components of rod-mediated dark adaptation; time to baseline sensitivity; and baseline (prebleach) scotopic sensitivity. RESULTS: Although their visual acuity was at least 20/25, patients with early ARM on average exhibited deficits in almost all rod-mediated parameters of dark adaptation as compared with age-similar healthy participants. For example, the rod-cone break was delayed approximately 10 minutes in early ARM patients as compared with healthy participants. Age-related maculopathy patients were more likely to fall outside the normal reference range for variables representing dark adaptation kinetics than for steady-state visual functions such as scotopic sensitivity. For example, 85% of ARM patients fell outside the normal reference range in at least one dark adaptation kinetic parameter, whereas only 25% of ARM patients fell outside the normal reference range for steady-state scotopic sensitivity. CONCLUSIONS: Rod-mediated kinetic parameters of dark adaptation, which reflect the sensitivity recovery of the visual cycle, are disrupted early in ARM pathogenesis. PMID- 11425676 TI - Induced corneal astigmatism after macular translocation surgery with scleral infolding. AB - OBJECTIVE: To document the corneal astigmatism that occurs with macular translocation after scleral infolding surgery. DESIGN: Retrospective case series of a nonrandomized clinical trial. PARTICIPANTS: Eight consecutive age-related macular degeneration patients (eight eyes) with choroidal neovascularization who underwent macular translocation with scleral infolding at the Duke University Eye Center from December 1998 through October 1999. METHODS: We retrospectively reviewed the charts of eight consecutive patients who underwent macular translocation surgery involving scleral infolding in the superotemporal quadrant. Two patients subsequently underwent release of scleral infolding. MAIN OUTCOME MEASURES: After surgery, these eyes were evaluated for corneal astigmatism with manifest refraction, keratometry, and computerized corneal topography. RESULTS: All eight eyes of eight patients revealed marked degrees of corneal astigmatism. Measurement of astigmatism via manifest refraction, keratometry, and corneal topography confirmed postoperative astigmatism corresponding to the axis of the scleral infolding. The amount of corneal astigmatism ranged from 1.75 to 7.37 diopters (D; mean, 4.60 D), with steepening along the axis of scleral infolding in the superotemporal quadrant of each eye (mean, 42.50 degrees from vertical; range, 24 degrees -66 degrees from vertical). Release of scleral infolding in two patients resulted in significant reduction of corneal astigmatism. CONCLUSIONS: Scleral shortening procedures used in macular translocation surgery may induce large amounts of corneal astigmatism. These patients should be assessed with keratometry and corneal topography to determine the accurate amount and axis. Thereafter, contact lens fitting or scleral infolding release may be considered as therapeutic options for large amounts of astigmatism persisting after surgery. PMID- 11425677 TI - Amniotic membrane transplantation in entropion surgery. AB - PURPOSE: To evaluate the role of amniotic membrane transplantation in the management of cicatricial eyelid entropion. DESIGN: Prospective, noncomparative interventional case series. PARTICIPANTS: Eighteen consecutive patients with cicatricial entropion. METHODS: A gray line lid split procedure with vertical anterior lamella repositioning was performed on 25 eyelids (upper or lower) of 18 patients with moderate to severe cicatricial entropion. Preserved human amniotic membrane (AM) was used to cover the bare tarsus up to the lid margin and secured with running 7-0 Vicryl. Impression cytology of the AM was performed at various stages postoperatively to study the epithelialization process. MAIN OUTCOME MEASURES: (1) Reepithelialization of bare tarsus, (2) extent of tarsal shrinkage, (3) recurrence of entropion. RESULTS: All the AM grafts took well. The most common complication was hemorrhage below the graft, which occurred in six cases. Complete success with no lashes touching the globe was achieved in 22 of 25 (88%) lids after a minimum follow-up of 12.0 months. The mean follow-up was 17.8 months. Two cases (qualified success) had recurrent trichiasis treated successfully with electrolysis. One case with severe trachomatous upper lid entropion recurred 14 months after surgery. The AM accelerated the epithelialization of bare tarsus; this was demonstrated by lack of fluorescein staining and reversion to skin color within 2 to 3 weeks. However, AM could not prevent tarsal shrinkage. Impression cytology demonstrated that features of conjunctival epithelium were present for the first postoperative month, but this was gradually replaced by squamous metaplasia, with keratinization appearing as early as 3 weeks postoperatively. CONCLUSIONS: The use of AM in a lid split procedure for correction of cicatricial entropion helps the bare tarsus epithelialize rapidly and improves the initial cosmetic result of surgery. PMID- 11425678 TI - Amniotic membrane transplantation in the management of shield ulcers of vernal keratoconjunctivitis. AB - PURPOSE: To report our experience with amniotic membrane transplantation in the management of severe shield ulcers. DESIGN: Retrospective, interventional, noncomparative case series. PARTICIPANTS: Four patients (seven eyes) with grade 2 (ulcer with opaque base) and grade 3 (plaquelike lesions) shield ulcers not responding to steroid therapy with or without surgical debridement. INTERVENTION: Amniotic membrane transplantation with stromal side down was performed after meticulous debridement of the ulcer. MAIN OUTCOME MEASURES: Healing of the ulcer with no epithelial defect. RESULTS: The ulcers healed with disintegration or retraction of the membrane in all patients within 2 weeks. CONCLUSIONS: Amniotic membrane transplantation in combination with debridement is an effective surgical modality in the management of severe shield ulcers. Further studies are warranted to confirm the efficacy of amniotic membrane transplantation in the management of shield ulcer and its correct position in the treatment algorithm. PMID- 11425679 TI - An outbreak of trematode-induced granulomas of the conjunctiva. AB - PURPOSE: To describe the epidemiologic, clinical, and histopathologic features of trematode granulomas of the conjunctiva, eyelid, and anterior chamber in pediatric patients. DESIGN: Prospective noncomparative case series. PARTICIPANTS: Forty-one children from a southern Indian village with conjunctival granulomas. METHODS: The village of Sellananthal was selected for a field visit after analysis of earlier hospital-based allergic conjunctival granuloma cases. Children with ocular diseases were examined, and histories of exposure to assumed risk factors and clinical findings were evaluated. Selected patients were brought to the base hospital for excisional biopsy. Serial sections obtained from the excised nodules were examined for the presence of a parasite. MAIN OUTCOME MEASURES: Histopathologic examination of excised conjunctival lesions or response of lesions to local medical therapy. RESULTS: In this year-long prospective study, 41 children (16 years or younger; 38 boys and 3 girls) with clinical features of allergic conjunctival granulomas were examined. Thirty-four patients were from a single village located in the southern Indian state of Tamil Nadu; the remaining 7 were from various parts of the same state. All children swam in their village's freshwater pond. Twenty patients with nodules less than 5 mm in diameter received medical treatment; 13 with larger nodules underwent surgical excision of the lesions. Nine of these 13 cases revealed a zonal granulomatous inflammation admixed with eosinophilic leukocytes; 4 of these 9 displayed fragments of the tegument and internal structures of a trematode and Splendore Hoeppli phenomenon. The remaining 4 of the 13 cases revealed nongranulomatous inflammation made up of lymphocytes, histiocytes, and eosinophils. Eight patients refused surgical treatment. CONCLUSIONS: In southern India, one cause of allergic conjunctival granulomas in children seems to be trematode infection. The clustering of cases in a single village and exposure to a village freshwater pond indicate the need for an epidemiologic investigation and study of the parasite's life cycle. Sporadic cases from other parts of the state with similar histories of exposure to their local pond or river water suggest a widespread distribution of the etiologic agent. PMID- 11425680 TI - Effects of laser in situ keratomileusis on tear production, clearance, and the ocular surface. AB - PURPOSE: To evaluate components of the integrated ocular surface/lacrimal gland unit in a series of patients before and after undergoing bilateral laser in situ keratomileusis (LASIK). DESIGN: Prospective, noncomparative case series. PARTICIPANTS: Forty-eight eyes of 14 men and 34 women (age range, 26-54; mean, 39.2 years) who underwent bilateral LASIK for myopia or myopic astigmatism. METHODS: LASIK was performed using a VISX Star Excimer Laser (Santa Clara, CA). Patients completed a questionnaire containing 11 questions that evaluated the character and severity of ocular irritation symptoms. Snellen visual acuity, tear fluorescein clearance, corneal fluorescein staining, aqueous tear production by the Schirmer 1 test, and corneal and conjunctival sensitivity were measured in each eye. Corneal surface regularity (SRI) was evaluated with the Tomey TMS-1 (Tomey, Cambridge, MA) topography instrument. Each randomly chosen eye was evaluated 1 to 2 days (T0) before LASIK and 7 days (T1), 1 (T2), 2 (T3), 6 (T4), 12 (T5), and 16 (T6) months postoperatively. A Wilcoxon test, two-tailed paired t test, Friedman test, or analysis of variance were used for statistical comparisons. MAIN OUTCOME MEASURES: Components of the integrated ocular surface/lacrimal gland unit. RESULTS: Both corneal and conjunctival sensitivity were noted to be significantly decreased from preoperative levels at 1week, 1 month, 12 months, and 16 months postoperatively (P < 0.0002 at each time point). Symptom severity scores were significantly increased at 1 week, 12 months, and 16 months postoperatively (P < 0.007 at all time points). The mean Schirmer 1 test scores were 24 +/- 14 mm preoperatively, and they decreased to 18 +/- 14 mm by 1 month postoperatively (P < 0.001). Tear fluorescein clearance showed a linear increase postoperatively and was significantly greater than baseline (P < 0.001) at each time point. There was a significant increase in punctate corneal fluorescein staining at 1 week postoperatively (P < 0.0001), but staining returned to baseline by 12 months. There was a statistically significant increase in SRI 1 week postoperatively (P < 0.007) with return to baseline levels by 6 months. CONCLUSIONS: Sensory denervation of the ocular surface after bilateral LASIK disrupts ocular surface tear dynamics and causes irritation symptoms. Patients undergoing LASIK should be informed of these risks. PMID- 11425681 TI - A prospective, randomized comparison of the use versus non-use of topical corticosteroids after laser in situ keratomileusis. AB - OBJECTIVE: To evaluate the benefit and safety of topical corticosteroids after laser in situ keratomileusis (LASIK). DESIGN: Prospective, randomized, double masked clinical trial. PARTICIPANTS: A consecutive series of 1747 eyes in 897 patients desiring surgical correction of myopia from plano to -16 diopters (D) with up to 6 D of astigmatism. INTERVENTION: Patients were randomly assigned to topical corticosteroids or artificial tears. MAIN OUTCOME MEASURES: The primary outcome measure was stability of refraction. Secondary outcome measures were differences in best-corrected visual acuity, decrease in spherical refractive myopia, improvement in uncorrected visual acuity, predictability of achieved corrections versus planned corrections, evaluation of postoperative haze, preservation of best-corrected visual acuity, intraocular pressure, and incidence of complications and adverse events. RESULTS: No difference was found between the groups for refractive accuracy, best-corrected or uncorrected visual acuity, or safety. Topical corticosteroids were associated with a prolonged period of refractive regression in moderate to high myopes (-7.25 to -16.00 D) (P = 0.01). Increased haze was noted at the 1 week examination for eyes not receiving topical steroids (P = 0.008); however, in most cases this was clinically insignificant. Thirty-six eyes (2.1%) required supplemental topical corticosteroids unrelated to reoperation. Two eyes received these on the first postoperative day and 15 at the 1-week examination. CONCLUSIONS: Topical corticosteroids did not seem to play any beneficial role in routine postoperative treatment after LASIK, and they were associated with decreased stability of refraction in more highly myopic eyes. Eyes that require additional topical corticosteroids are easily identified at either the initial postoperative examinations or during the course of routine postoperative care. PMID- 11425683 TI - Correction of irregular astigmatism with excimer laser assisted by sodium hyaluronate. AB - PURPOSE: To demonstrate the usefulness of excimer laser surgery assisted by sodium hyaluronate for the correction of irregular astigmatism after previous corneal refractive surgery. DESIGN: Prospective noncomparative case series. PARTICIPANTS: Fifty eyes with surgically induced irregular astigmatism. All the patients had been subjected previously to one or more of the following: laser in situ keratomileusis (LASIK), incisional keratotomy, photorefractive keratotomy, phototherapeutic keratotomy, laser thermokeratoplasty, and corneal trauma. Irregular astigmatism developed thereafter. METHODS: Stability of the corneal topography pattern before surgical decision was observed for at least 6 months with the C-SCAN corneal topography with Ray Tracing (Technomed GmbH, Germany) and the Eye-Sys 2000 Corneal Analysis System (Eye Sys Co., Houston, TX). The lowest pachymetry value was 310 microm in the thinnest portion of the cornea. Ablation was performed with the Technolas 217 C-LASIK excimer laser (Bausch and Lomb Chiron Technolas GmbH, Dornach, Germany), in phototherapeutic keratectomy mode, assisted by a mask of 0.25% sodium hyaluronate. We performed our ablation either on the surface or under a flap, whether elevated or newly cut. MAIN OUTCOME MEASURES: Uncorrected and best-corrected visual acuity, manifest and cycloplegic refraction, superficial corneal surface quality, image distortion, and predicted corneal visual acuity. RESULTS: At 3 months of follow-up, the irregular corneal surface was significantly improved in all cases (n = 50). The superficial corneal surface quality improved from 70.5% +/- 9.16% to 75.6% +/- 10.38 (P < 0.0001). Six eyes lost 1 line of best-corrected visual acuity, three eyes lost 2 lines, and five eyes lost 3 or more lines. The real corneal ablation depth obtained was equal to 63% of that programmed because of the influence of masking substance. Mean uncorrected visual acuity improved from 20/80 +/- 20/125 to 20/63 +/- 20/100 (P = 0.01). Predicted corneal visual acuity improved from a mean of 20/32 +/- 20/80 to 20/25 +/- 20/63 (P = 0.004). Image distortion improved from a mean of 13.95 +/- 3.64 to 12.16 +/- 3.92 (P < 0.0001). We obtained a hyperopic shift in 56% and myopic shift in 40% of eyes. After 6 months of follow-up the irregular corneal surface continued to improve in all cases (n = 32). The superficial corneal surface quality improved from 69.38% +/- 9.48% to 73.13% +/- 8.87 (P = 0.002). Two eyes lost 2 lines of best-corrected visual acuity, and 3 eyes lost 1 line. Ray tracing was significantly improved in all cases at the end of follow-up regarding superficial corneal surface quality (P = 0.002) and the image distortion (P = 0.05). Improvement of predicted corneal visual acuity was not significant (P = 0.11). The procedure proved to be safe, with a safety index of 1.1. Differences between the surface and stromal treatments and between pattern and nonpattern irregular astigmatism were not statistically significant. CONCLUSION: It is possible to produce a more regular corneal surface and to improve best-corrected visual acuity in patients with irregular astigmatism using plano-scan excimer laser assisted by viscous masking solution of 0.25% sodium hyaluronate. PMID- 11425684 TI - Effect of varying the optical zone diameter on the results of hyperopic laser in situ keratomileusis. AB - PURPOSE: To compare the predictability and safety of laser in situ keratomileusis (LASIK) for low to moderate spherical hyperopia using different ablation zone diameters. DESIGN: Retrospective, nonrandomized, comparative trial. PARTICIPANTS: Forty-nine eyes that underwent hyperopic LASIK. INTERVENTION: Two surgeons (JMD, RKM) performed hyperopic LASIK using the VISX STAR S2 excimer laser (VISX, Inc., Sunnyvale, CA) and the Bausch & Lomb Hansatome microkeratome (Chiron Vision, Irvine, CA) using ablation zone diameters of 5 x 9 mm, 5.5 x 8.5 mm, or 6 x 9 mm (the first number represents the optical zone diameter and the second number represents the diameter of the outer border of the ablation zone). MAIN OUTCOME MEASURES: Refractive and visual outcomes at 3 to 6 months after surgery were analyzed. Groups were compared for deviations from targeted spherical equivalent, uncorrected visual acuity, and loss of best spectacle-corrected visual acuity (BSCVA). RESULTS: The mean intended hyperopic correction was +2.48 +/- 1.13 diopters (D; 0.63-5.50 D). There were 16 eyes in the 5 x 9-mm group, 15 eyes in the 5.5 x 8.5-mm group, and 18 eyes in the 6 x 9-mm group. On average, the 5 x 9 mm group achieved 97% of the programmed correction, the 5.5 x 8.5-mm group achieved 104%, and the 6 x 9-mm group achieved 112% of the programmed correction. The tendency toward overcorrection in the 6 x 9-mm group compared with the 5 x 9 mm group was statistically significant (P < 0.05). The incidence of one line loss of BSCVA was greatest in the 5 x 9-mm group (19%) and lowest in the 6 x 9-mm group (6%). These differences were not statistically significant. No eyes experienced a loss of two or more lines of BSCVA at last examination. CONCLUSIONS: Hyperopic LASIK using the VISX STAR is safe and effective using different ablation zone diameters. There appears to be an increased tendency toward overcorrection with progressively larger optical zone diameters. PMID- 11425685 TI - Infectious keratitis after laser refractive surgery. AB - PURPOSE: To report two cases of infectious keratitis, one fungal after photorefractive keratectomy (PRK) and the other bacterial after laser in situ keratomileusis (LASIK). DESIGN: Two interventional case reports. PARTICIPANTS: Case 1 is a male who was seen 3 weeks after PRK with a corneal ulceration. Case 2 involves a female who was seen 7 weeks after LASIK with interface granularity. RESULTS: Cultures in case 1 were identified as Scopulariopsis species, and despite intensive treatment, a therapeutic penetrating keratoplasty (PK) was eventually performed. Case 2 had cultures identified as Mycobacterium chelonae and also ultimately required a therapeutic PK. CONCLUSIONS: Two unusual infectious keratitides are reported after different laser refractive surgery techniques. PMID- 11425686 TI - Factors that influence the surgical effects of astigmatic keratotomy after cataract surgery. AB - OBJECTIVE: To determine the factors affecting the surgical effect of astigmatic keratotomy (AK) when against-the-rule astigmatism is present following cataract surgery. DESIGN: Prospective interventional noncomparative case series. PARTICIPANTS: Twenty eyes of 19 patients from four medical centers who had against-the-rule astigmatism following cataract surgery. INTERVENTION: AK with a 6 mm optical zone, two linear 3-mm length incisions (T-cut) and a depth of 90% of the central thickness was performed on all subjects. MAIN OUTCOME MEASURES: Vector analysis of astigmatic correction. Multiple regression analysis for seven covariates including age, spherical equivalent of the manifest refraction, preoperative astigmatism, corneal diameter, corneal thickness, mean radius of corneal curvature and axial misalignment. RESULTS: Multiple regression analysis showed that the preoperative astigmatism (p = 0.014) and the axis deviation (p = 0.005) were significantly correlated with the surgical effects. CONCLUSIONS: Even with a uniform surgical procedure, the surgical effects of AK in eyes with against-the-rule astigmatism can be affected by the amount of preoperative astigmatism and the intraoperative axis misalignment. Adding the amount of preoperative astigmatism to the nomogram and improvement of surgical procedures will be required to obtain better surgical predictability of AK following cataract surgery. PMID- 11425687 TI - Incidence of nonarteritic anterior ischemic optic neuropathy associated with cataract extraction. AB - OBJECTIVES: To determine the incidence of nonarteritic anterior ischemic optic neuropathy (NAION) after cataract extraction and to use the incidence to evaluate the causal relationship between cataract extraction and NAION. DESIGN: Cohort study. PARTICIPANTS: Five thousand seven hundred eighty-seven cataract extraction cases at the Bascom Palmer Eye Institute (BPEI) in a 5-year period between January 1, 1993 and December 31, 1997. METHODS: During the study period, an estimated 5787 cataract extraction cases were identified that met the following criteria: (1) the patient was 50 years of age or older, (2) surgery was performed by phacoemulsification or phacofracture techniques and not planned in combination with another procedure, and (3) follow-up was at BPEI. Charts of patients diagnosed with NAION from January 1, 1993 to December 31, 1998 were reviewed to detect the occurrence of NAION within 1 year of surgery. The diagnosis of NAION was based on the following: (1) an acute decrease in vision, (2) associated nerve fiber layer defect on visual field testing, (3) associated relative afferent pupillary defect, and (4) observed optic nerve edema. The exact binomial test was used to compare the incidence of NAION after cataract extraction against the expected incidence. MAIN OUTCOME MEASURE: Occurrence of NAION within 1 year after cataract extraction. RESULTS: Of the 5787 cataract extraction cases, three patients experienced NAION within 1 year of the procedure. Each case occurred after surgery on days 29, 36, and 117, giving an estimated 6-month incidence of 51.8 in 100,000 and a 6-week incidence of 34.6 in 100,000. Both were statistically higher than the previously reported overall incidence of NAION. Two of the three patients had a history of NAION in the contralateral eye. CONCLUSIONS: Cataract extraction is associated with an increased incidence of NAION, which is higher than the reported overall incidence. However, the risk of NAION after cataract extraction is low, with approximately one occurrence in every 2000 cases. A history of NAION in the fellow eye may be a risk factor. PMID- 11425688 TI - Effects of rigid and soft contact lens daily wear on corneal epithelium, tear lactate dehydrogenase, and bacterial binding to exfoliated epithelial cells. AB - OBJECTIVE: To determine the effects of lens type and oxygen transmissibility on human corneal epithelium during daily lens wear (DW). DESIGN: Prospective, randomized, double-masked, single-center, parallel treatment groups clinical trial. PARTICIPANTS: Two hundred forty-six patients fitted with: (1) high oxygen transmissible soft lenses (n = 36), (2) hyper oxygen-transmissible soft lenses (n = 135), and (3) hyper oxygen-transmissible rigid gas-permeable (RGP) lenses (n = 75). INTERVENTION: Irrigation chamber to collect exfoliated epithelial surface cells, confocal microscopy, and tear collection at baseline, 2 weeks, and 4 weeks of DW. MAIN OUTCOME MEASURES: (1) Pseudomonas aeruginosa (PA) binding to exfoliated corneal epithelial surface cells, (2) central epithelial thickness, (3) superficial epithelial cell area, (4) epithelial surface cell exfoliation, and (5) tear lactate dehydrogenase (LDH). RESULTS: Four weeks of DW with the high oxygen-transmissible soft lens significantly increased PA binding from baseline 6.55 +/- 3.01 to 8.75 +/- 3.05 bacteria per epithelial cell (P < 0.01). By contrast, hyper oxygen-transmissible soft lens wear increased binding significantly less (6.13 +/- 2.45 to 7.62 +/- 3.06; P < 0.01), whereas hyper oxygen-transmissible RGP lens wear demonstrated no significant changes (5.91 +/- 2.40 to 6.13 +/- 2.17; P = 0.533). No significant change in central epithelial thickness was found after 4 weeks of DW in either soft lens; however, the epithelial thickness decreased by 9.8% (P < 0.001) with RGP lens wear. Epithelial cell surface area increased 3.3% and 4.1% with the high and hyper oxygen transmissible soft lenses, respectively, and 10.5% with the hyper oxygen transmissible RGP lens (P < 0.001). Epithelial desquamation significantly decreased in all groups (P < 0.001). Tear LDH levels increased for all test lenses (P < 0.001). CONCLUSIONS: Increased PA binding induced by wear of a conventional soft lens material is significantly greater than that induced by the new hyper oxygen-transmissible soft silicone hydrogel lens during DW. However, both soft materials showed significant increases in PA binding as compared with baseline controls. By contrast, hyper oxygen-transmissible RGP lens DW did not increase PA binding significantly. Taken together, these findings suggest for the first time both an oxygen effect as well as a difference between soft and rigid lens types on PA binding in DW. PMID- 11425689 TI - Secondary ocular involvement in systemic "memory" B-cell lymphocytic leukemia. AB - OBJECTIVE: Recently, B-cell chronic lymphocytic leukemia (B-CLL) has been subdivided into "naive" B-CLL and "memory" B-CLL on the basis of the presence of somatic mutations in the variable region of the immunoglobulin heavy chain gene (IgH). The aim of the current paper was to report the clinical, histopathologic, and molecular biologic findings of intraocular and ocular adnexal involvement in a patient with systemic B-CLL. DESIGN: Case report. INTERVENTION: Treatment consisted of systemic chemotherapy, conjunctival biopsies, and, ultimately, enucleation of the left eye. METHODS: Histopathologic findings of a bone marrow biopsy, the conjunctival biopsies, and the enucleated eye were compared. Further, extensive immunohistochemistry, polymerase chain reaction (PCR) for the detection of heavy chain (IgH) gene rearrangement, gene scan analysis, and DNA sequencing were performed on all tissues. RESULTS: The tumor manifestations in all specimens demonstrated similar morphologic and immunophenotypic characteristics, consistent with the diagnosis of B-CLL. Immunoglobulin-H PCR and gene scan analysis showed that the B-CLL infiltrates consisted of B cells derived from the same clone. DNA sequencing demonstrated the presence of eight somatic mutations in the variable region of IgH, consistent with "memory" B-CLL. CONCLUSIONS: Secondary ocular manifestations of B-CLL occur relatively commonly during disease progression. In the current case of memory B-CLL, ocular manifestation of the disease occurred 16 years after initial diagnosis, agreeing with clinical studies suggesting that a less aggressive course is seen in "memory" B-CLL than its counterpart, "naive" B CLL. Somatic mutation analysis in the variable region of IgH in B-CLL should be part of routine staging investigations to aid the prediction of the individual clinical course in B-CLL patients and to determine new therapeutic strategies. PMID- 11425690 TI - Surgical excision of iris nodules in the management of sarcoid uveitis. AB - PURPOSE: To illustrate the role of surgical removal of iris nodules (granulomas) in the management of sarcoid uveitis. STUDY DESIGN: Two interventional case reports. METHODS: The authors describe the clinical course of two 10-year-old males with long-standing granulomatous uveitis refractory to medical antiinflammatory and immunomodulatory therapy. Both patients were seen with iris masses, which were excised and biopsied, with findings of sarcoidosis. MAIN OUTCOME MEASURES: Control of ocular inflammation as evidenced by decrease in inflammatory cells in both anterior and posterior chambers. RESULTS: On excision of the iris masses, the ocular inflammation was controlled in both patients. CONCLUSIONS: The authors hypothesize that iris granulomas may not only be products of persistent antigenic stimulation characteristic of sarcoidosis but subsequently become foci of continued cytokine production and ocular inflammation. Total surgical removal of the iris masses may help in the diagnosis and control of sarcoid uveitis refractory to medical management. PMID- 11425691 TI - Photographic and clinical techniques for outcome assessment in sixth nerve palsy. AB - PURPOSE: There are few data on the reproducibility of clinical assessment of ductions and alignment. We evaluated photographic methods that may be used for masked outcome determination in a clinical trial and compared them with clinical measures. DESIGN: Interexaminer reliability study. PARTICIPANTS: Twenty-three patients with unilateral sixth nerve palsy and three control participants were clinically evaluated by two masked examiners. MAIN OUTCOME MEASURES: Abduction deficit was graded as 0 to -5. Simultaneous prism and cover tests (SPCT) and alternate cover tests (ACT) were performed at distance and near fixation. Photographs were taken of abduction and distance alignment by each examiner. The photographs were evaluated by a third masked reader, who assigned abduction grade, measured absolute abduction (mm) and relative abduction (%), and calculated alignment in prism diopters (pd). Agreement was evaluated by calculating intraclass correlation coefficients (r(i)), weighted kappa statistics (kappa), and Spearman rank correlation coefficients (r(S)). RESULTS: There was excellent agreement between the two clinicians in clinical abduction deficit (kappa = 0.86) SPCT and ACT at distance and near (r(i) 0.94-0.96), between the clinical grade and masked photographic grade (kappa = 0.83), and between the two sets of photographs for absolute abduction and relative abduction (r(i) = 0.98 and 0.97). Both photographic measures of abduction correlated well with the clinical grade (r(S) = -0.96 for each). Measurements of alignment from photographs correlated with clinical SPCT measurement (r(i) = 0.88), but had a lower level of absolute agreement (38% within 5 pd) than between two independent SPCT measurements (96% within 5 pd). CONCLUSIONS: The excellent interexaminer agreement of our new photographic abduction assessment and of masked clinical measures suggest that these methods would be useful in clinical trials. In contrast, our simple method of photographic assessment of alignment lacks excellent agreement with the clinical assessment. These data are important in planning clinical trials in strabismus. PMID- 11425692 TI - Lacrimal sac dacryoliths: predictive factors and clinical characteristics. AB - OBJECTIVE: Lacrimal sac dacryoliths are often diagnosed during dacryocystorhinostomy (DCR), although their cause is unclear. Several factors have been suggested to predispose to dacryolith formation. The clinical presentation of nasolacrimal duct obstruction (NLDO) may differ if associated with a dacryolith. Our study evaluated specific risk factors related to dacryolith formation and how the clinical presentation of patients with dacryoliths differs from patients with primary acquired NLDO who undergo DCR. DESIGN: Retrospective, comparative, interventional case series. PARTICIPANTS/INTERVENTION: One hundred thirty-eight consecutive patients with NLDO who underwent DCR (163 total DCR cases) between 1993 and 1998. MAIN OUTCOME MEASURES: We evaluated the frequency of dacryolith formation for all DCR cases. For 115 patients with primary acquired NLDO, we statistically compared the patients with dacryoliths (n = 12) to those without (n = 103) for several variables related to clinical history and presentation. RESULTS: The overall frequency of dacryoliths was 12 of 163 (7.4%) for all DCR cases and 12 of 138 (8.7%) for all patients. All patients with dacryoliths were in the subgroup of 115 patients with primary acquired NLDO, and the frequency in that subgroup was 12 of 115 (10.4%). There was no statistical difference between the group of patients with dacryoliths and those without dacryoliths for age, duration of epiphora, history of acute dacryocystitis, or previous use of antiglaucomatous topical medications. Male gender was more likely to be associated with dacryoliths (P = 0.004), as was initial presentation with lacrimal sac distension (P < 0.001). Partial nasolacrimal obstruction on lacrimal irrigation approached statistical significance for dacryolith formation (P = 0.08), as did a history of cigarette smoking (P = 0.09). CONCLUSIONS: In patients with primary acquired NLDO who require DCR, male gender and presence of sac distension are more frequently associated with dacryoliths. Partial NLDO and history of cigarette smoking may also be relative risk factors for dacryolith formation. These observations may be helpful in the evaluation and surgical planning for patients with lacrimal obstruction. PMID- 11425694 TI - Congenital fibrosis of the extraocular muscles associated with cortical dysplasia and maldevelopment of the basal ganglia. AB - BACKGROUND: Congenital fibrosis of the extraocular muscles (CFEOM) is a rare condition that has been traditionally regarded as a primary eye muscle disease. Recent studies, however, suggest that CFEOM may be the result of a primary neuropathy with secondary myopathic changes. PURPOSE: To describe a previously unrecognized association between congenital fibrosis of the extraocular muscles and structural abnormalities of the brain. DESIGN: Small case series. METHODS: Detailed clinical examinations and neuroradiologic studies were performed on the three affected family members. In addition, genetic analysis of the family was performed. RESULTS: The three affected family members, mother and two children, have the ocular features of 'classic' congenital fibrosis of the extraocular muscles. All showed dilation of the left lateral ventricle secondary to hypoplasia of the body and tail of the ipsilateral caudate nucleus. There was fusion of an enlarged caudate nucleus head with the underlying putamen. Both children showed widespread bilateral cortical dysplasia. Genetic analysis of the family was inconclusive but consistent with linkage to the CFEOM1 locus on chromosome 12. Chromosomal analysis of the affected individuals did not show evidence of a deletion of chromosome 12 and haplotype analysis was not suggestive of a microdeletion. CONCLUSIONS: Cerebral cortical and basal ganglia maldevelopment can be found in individuals with CFEOM. This suggests that neuroimaging should be considered in the initial diagnostic evaluation of these patients, particularly if there is developmental delay. PMID- 11425695 TI - Late endophthalmitis associated with glaucoma drainage implants. AB - OBJECTIVE: To report the clinical course of a series of patients who had late endophthalmitis develop after glaucoma drainage implant (GDI) surgery. DESIGN: Noncomparative, interventional, consecutive case series. PARTICIPANTS: Four patients were identified with late endophthalmitis associated with Baerveldt glaucoma implants. METHODS: The medical records of all patients with endophthalmitis associated with a GDI treated at the Bascom Palmer Eye Institute or University of Florida between January 1, 1987 and December 31, 1999 were retrospectively reviewed. Patients with culture-positive endophthalmitis diagnosed more than 1 month after GDI surgery were included in this series. MAIN OUTCOME MEASURES: Visual acuity and intraocular pressure (IOP). RESULTS: Late endophthalmitis associated with Baerveldt glaucoma implants developed 7 weeks to 2 years postoperatively. Exposure of the GDI tube was present in all cases. The implant was removed in three of four patients. Visual acuity worsened from preinfection level in two of four cases. IOP was controlled at last follow-up in all patients, although replacement of the explanted GDI was required in one patient. CONCLUSIONS: Late endophthalmitis may occur after GDI surgery. Exposure of the GDI tube seems to represent a major risk factor for these infections. To prevent this potentially devastating complication, we recommend prophylactic surgical revision with a patch graft in all cases in which there is an exposed GDI tube. PMID- 11425696 TI - Surgical management of macular holes: a report by the American Academy of Ophthalmology. AB - OBJECTIVE: The document describes macular hole surgery and examines the available evidence to address questions about the efficacy of the procedure for different stages of macular hole, complications during and after surgery, and modifications to the technique. METHODS: A literature search conducted for the years 1968 to 2000 retrieved over 400 citations that matched the search criteria. This information was reviewed by panel members and a methodologist, and it was evaluated for the quality of the evidence presented. RESULTS: There are three multicenter, controlled, randomized trials that constitute Level I evidence and compare the value of surgery versus observation for macular hole. There are three multicenter, controlled, randomized trials studying the use of adjuvant therapy in macular hole repair. Postoperative vision of 20/40 or better has been reported in 22% to 49% of patients in randomized trials. The risks of surgical complications include retinal detachment (3%), endophthalmitis (<1%), cataract (>75%), and late reopening the hole (2% to 10%). CONCLUSIONS: The evidence does not support surgery for patients with stage 1 holes. Level I evidence supports surgery for stage 2 holes to prevent progression to later stages of the disease and further visual loss. Level I evidence shows that surgery improves the vision in a majority of patients with stage 3 and stage 4 holes. There is no strong evidence that adjuvant therapy used at the time of surgery results in improved surgical outcomes. Patient inconvenience, patient preference, and quality of life issues have not been studied. PMID- 11425697 TI - Oxidation of reduced inorganic sulfur compounds by bacteria: emergence of a common mechanism? PMID- 11425699 TI - Antacid increases survival of Vibrio vulnificus and Vibrio vulnificus phage in a gastrointestinal model. AB - Viable counts of three strains of Vibrio vulnificus and its phage were determined during exposure to a mechanical gastrointestinal model with or without antacid for 9 h at 37 degrees C. V. vulnificus was eliminated (>4-log reduction) within 30 min in the gastric compartment (pH decline from 5.0 to 3.5). Viable V. vulnificus cells delivered from the gastric compartment during the first 30 min of exposure reached 10(6) to 10(8) CFU/ml in the intestinal compartment after 9 h (pH 7.0). Phages were eliminated within 45 min in the gastric compartment (pH decline from 5.1 to 2.5). Less than a 2-log reduction of phage was observed in the intestinal compartment after 9 h (pH 7.0). When the gastric compartment contained antacid V. vulnificus counts decreased slightly (<2 log) during 2 h of exposure (pH decline from 7.7 to 6.0), while counts in the intestinal compartment (pH 7.5) reached 10(7) to 10(9) CFU/ml. Phage numbers decreased 1 log after 2 h in the gastric compartment (pH decline from 7.7 to 5.7) containing antacid and decreased 1 log in the intestinal compartment (pH 7.6) after 9 h. Presence of antacid in the gastric compartment of the model greatly increased the ability of both V. vulnificus and its phage to survive simulated gastrointestinal transit and may be a factor involved with oyster-associated illness. PMID- 11425698 TI - Antifungal proteins. PMID- 11425700 TI - The functional ccpA gene is required for carbon catabolite repression in Lactobacillus plantarum. AB - We report the characterization of the ccpA gene of Lactobacillus plantarum, coding for catabolite control protein A. The gene is linked to the pepQ gene, encoding a proline peptidase, in the order ccpA-pepQ, with the two genes transcribed in tandem from the same strand as distinct transcriptional units. Two ccpA transcription start sites corresponding to two functional promoters were found, expression from the upstream promoter being autogenously regulated through a catabolite-responsive element (cre) sequence overlapping the upstream +1 site. During growth on ribose, the upstream promoter showed maximal expression, while growth on glucose led to transcription from the downstream promoter. In a ccpA mutant strain, the gene was transcribed mainly from the upstream promoter in both repressing and non repressing conditions. Expression of two enzyme activities, beta-glucosidase and beta-galactosidase, was relieved from carbon catabolite repression in the ccpA mutant strain. In vivo footprinting analysis of the catabolite-controlled bglH gene regulatory region in the ccpA mutant strain showed loss of protection of the cre under repressing conditions. PMID- 11425701 TI - Quantitative detection of Escherichia coli O157 in surface waters by using immunomagnetic electrochemiluminescence. AB - A protocol for the quantitative detection of Escherichia coli O157 in raw and concentrated surface waters using immunomagnetic electrochemiluminescence (IM ECL) was developed and optimized. Three antibody sandwich formats were tested: commercial anti-O157:H7 IM beads, IM beads made in-house with a polyclonal anti O157:H7 immunoglobulin G (IgG), or IM beads made in-house with a monoclonal anti O157:H7 IgG coupled with a polyclonal anti-O157:H7 IgG to which an electrochemiluminescent label (TAG) was attached. The monoclonal IM bead polyclonal TAG format was chosen for optimization because it gave lower background levels and linear regression slopes of ca. 1.0, indicative of a constant ECL signal per cell. The dynamic range was ca. 10(1) to 10(5) cells ml( 1) in phosphate-buffered saline and in raw water samples. The monoclonal IM beads selectively captured E. coli O157 cells in the presence of ca. 10(8) cells of a non-O157 strain of E. coli ml(-1). Background ECL signals from concentrated (100 fold) water samples were substantially higher and more variable than raw water samples. The background signal was partially eliminated by the addition of polyvinylpolypyrrolidone. Successive cell capture incubations, termed sequential bead capture (SBC), were optimized for establishing baseline ECL values for individual water samples. The linear dynamic range with SBC was ca. 10(2) to 10(5) E. coli O157 cells ml of concentrated water(-1). To validate the protocol, 10-liter surface water samples were spiked with ca. 5,000 E. coli O157 (Odwalla) cells and concentrated by vortex filtration, and 1- or 3-ml aliquots were analyzed by IM-ECL. Differential ECL signals (SBC) from 1- and 3-ml samples were statistically significant and were generally consistent with standard curves for these cell concentrations. Enrichments were conducted with aliquots of spiked raw water and concentrated water using EC broth and minimal lactose broth (MLB). All tubes with concentrated water became turbid and gave a positive ECL response for E. coli O157 (>10,000 ECL units); MLB gave a somewhat higher detection rate with spiked raw water. The potential sensitivity of the IM-ECL assay is ca. 25 E. coli O157 cells ml of raw water(-1), 25 cells 100 ml of 100-fold concentrated water( 1), or 1 to 2 viable cells liter(-1) with concentration and enrichment. The IM ECL assay appears suitable for routine analysis and screening of water samples. PMID- 11425702 TI - The leucine incorporation method estimates bacterial growth equally well in both oxic and anoxic lake waters. AB - Bacterial biomass production is often estimated from incorporation of radioactively labeled leucine into protein, in both oxic and anoxic waters and sediments. However, the validity of the method in anoxic environments has so far not been tested. We compared the leucine incorporation of bacterial assemblages growing in oxic and anoxic waters from three lakes differing in nutrient and humic contents. The method was modified to avoid O(2) contamination by performing the incubation in syringes. Isotope saturation levels in oxic and anoxic waters were determined, and leucine incorporation rates were compared to microscopically observed bacterial growth. Finally, we evaluated the effects of O(2) contamination during incubation with leucine, as well as the potential effects of a headspace in the incubation vessel. Isotope saturation occurred at a leucine concentration of above about 50 nM in both oxic and anoxic waters from all three lakes. Leucine incorporation rates were linearly correlated to observed growth, and there was no significant difference between oxic and anoxic conditions. O(2) contamination of anoxic water during 1-h incubations with leucine had no detectable impact on the incorporation rate, while a headspace in the incubation vessel caused leucine incorporation to increase in both anoxic and O(2) contaminated samples. The results indicate that the leucine incorporation method relates equally to bacterial growth rates under oxic and anoxic conditions and that incubation should be performed without a headspace. PMID- 11425703 TI - Photosynthetic and phylogenetic primers for detection of anoxygenic phototrophs in natural environments. AB - Primer sets were designed to target specific 16S ribosomal DNA (rDNA) sequences of photosynthetic bacteria, including the green sulfur bacteria, the green nonsulfur bacteria, and the members of the Heliobacteriaceae (a gram-positive phylum). Due to the phylogenetic diversity of purple sulfur and purple nonsulfur phototrophs, the 16S rDNA gene was not an appropriate target for phylogenetic rDNA primers. Thus, a primer set was designed that targets the pufM gene, encoding the M subunit of the photosynthetic reaction center, which is universally distributed among purple phototrophic bacteria. The pufM primer set amplified DNAs not only from purple sulfur and purple nonsulfur phototrophs but also from Chloroflexus species, which also produce a reaction center like that of the purple bacteria. Although the purple bacterial reaction center structurally resembles green plant photosystem II, the pufM primers did not amplify cyanobacterial DNA, further indicating their specificity for purple anoxyphototrophs. This combination of phylogenetic- and photosynthesis-specific primers covers all groups of known anoxygenic phototrophs and as such shows promise as a molecular tool for the rapid assessment of natural samples in ecological studies of these organisms. PMID- 11425704 TI - Assessing the risk of primary amoebic meningoencephalitis from swimming in the presence of environmental Naegleria fowleri. AB - Free-living Naegleria fowleri amoebae cause primary amoebic meningoencephalitis (PAM). Because of the apparent conflict between their ubiquity and the rarity of cases observed, we sought to develop a model characterizing the risk of PAM after swimming as a function of the concentration of N. fowleri. The probability of death from PAM as a function of the number of amoebae inhaled is modeled according to results obtained from animals infected with amoeba strains. The calculation of the probability of inhaling one or more amoebae while swimming is based on a double hypothesis: that the distribution of amoebae in the water follows a Poisson distribution and that the mean quantity of water inhaled while swimming is 10 ml. The risk of PAM for a given concentration of amoebae is then obtained by summing the following products: the probability of inhaling n amoebae x the probability of PAM associated with inhaling these n amoebae. We chose the lognormal model to assess the risk of PAM because it yielded the best analysis of the studentized residuals. Nonetheless, the levels of risk thereby obtained cannot be applied to humans without correction, because they are substantially greater than those indicated by available epidemiologic data. The curve was thus adjusted by a factor calculated with the least-squares method. This provides the PAM risk in humans as a function of the N. fowleri concentration in the river. For example, the risk is 8.5 x 10(-8) at a concentration of 10 N. fowleri amoebae per liter. PMID- 11425705 TI - Study of genetic diversity of eukaryotic picoplankton in different oceanic regions by small-subunit rRNA gene cloning and sequencing. AB - Very small eukaryotic organisms (picoeukaryotes) are fundamental components of marine planktonic systems, often accounting for a significant fraction of the biomass and activity in a system. Their identity, however, has remained elusive, since the small cells lack morphological features for identification. We determined the diversity of marine picoeukaryotes by sequencing cloned 18S rRNA genes in five genetic libraries from North Atlantic, Southern Ocean, and Mediterranean Sea surface waters. Picoplankton were obtained by filter size fractionation, a step that excluded most large eukaryotes and recovered most picoeukaryotes. Genetic libraries of eukaryotic ribosomal DNA were screened by restriction fragment length polymorphism analysis, and at least one clone of each operational taxonomic unit (OTU) was partially sequenced. In general, the phylogenetic diversity in each library was rather great, and each library included many different OTUs and members of very distantly related phylogenetic groups. Of 225 eukaryotic clones, 126 were affiliated with algal classes, especially the Prasinophyceae, the Prymnesiophyceae, the Bacillariophyceae, and the Dinophyceae. A minor fraction (27 clones) was affiliated with clearly heterotrophic organisms, such as ciliates, the chrysomonad Paraphysomonas, cercomonads, and fungi. There were two relatively abundant novel lineages, novel stramenopiles (53 clones) and novel alveolates (19 clones). These lineages are very different from any organism that has been isolated, suggesting that there are previously unknown picoeukaryotes. Prasinophytes and novel stramenopile clones were very abundant in all of the libraries analyzed. These findings underscore the importance of attempts to grow the small eukaryotic plankton in pure culture. PMID- 11425706 TI - Application of denaturing gradient gel electrophoresis (DGGE) to study the diversity of marine picoeukaryotic assemblages and comparison of DGGE with other molecular techniques. AB - We used denaturing gradient gel electrophoresis (DGGE) to study the diversity of picoeukaryotes in natural marine assemblages. Two eukaryote-specific primer sets targeting different regions of the 18S rRNA gene were tested. Both primer sets gave a single band when used with algal cultures and complex fingerprints when used with natural assemblages. The reproducibility of the fingerprints was estimated by quantifying the intensities of the same bands obtained in independent PCR and DGGE analyses, and the standard error of these estimates was less than 2% on average. DGGE fingerprints were then used to compare the picoeukaryotic diversity in samples obtained at different depths and on different dates from a station in the southwest Mediterranean Sea. Both primer sets revealed significant differences along the vertical profile, whereas temporal differences at the same depths were less marked. The phylogenetic composition of picoeukaryotes from one surface sample was investigated by excising and sequencing DGGE bands. The results were compared with an analysis of a clone library and a terminal restriction fragment length polymorphism fingerprint obtained from the same sample. The three PCR-based methods, performed with three different primer sets, revealed very similar assemblage compositions; the same main phylogenetic groups were present at similar relative levels. Thus, the prasinophyte group appeared to be the most abundant group in the surface Mediterranean samples as determined by our molecular analyses. DGGE bands corresponding to prasinophytes were always found in surface samples but were not present in deep samples. Other groups detected were prymnesiophytes, novel stramenopiles (distantly related to hyphochytrids or labyrinthulids), cryptophytes, dinophytes, and pelagophytes. In conclusion, the DGGE method described here provided a reasonably detailed view of marine picoeukaryotic assemblages and allowed tentative phylogenetic identification of the dominant members. PMID- 11425707 TI - Autotrophic ammonia oxidation at low pH through urea hydrolysis. AB - Ammonia oxidation in laboratory liquid batch cultures of autotrophic ammonia oxidizers rarely occurs at pH values less than 7, due to ionization of ammonia and the requirement for ammonium transport rather than diffusion of ammonia. Nevertheless, there is strong evidence for autotrophic nitrification in acid soils, which may be carried out by ammonia oxidizers capable of using urea as a source of ammonia. To determine the mechanism of urea-linked ammonia oxidation, a ureolytic autotrophic ammonia oxidizer, Nitrosospira sp. strain NPAV, was grown in liquid batch culture at a range of pH values with either ammonium or urea as the sole nitrogen source. Growth and nitrite production from ammonium did not occur at pH values below 7. Growth on urea occurred at pH values in the range 4 to 7.5 but ceased when urea hydrolysis was complete, even though ammonia, released during urea hydrolysis, remained in the medium. The results support a mechanism whereby urea enters the cells by diffusion and intracellular urea hydrolysis and ammonia oxidation occur independently of extracellular pH in the range 4 to 7.5. A proportion of the ammonia produced during this process diffuses from the cell and is not subsequently available for growth if the extracellular pH is less than 7. Ureolysis therefore provides a mechanism for nitrification in acid soils, but a proportion of the ammonium produced is likely to be released from the cell and may be used by other soil organisms. PMID- 11425708 TI - Detection of differential gene expression in biofilm-forming versus planktonic populations of Staphylococcus aureus using micro-representational-difference analysis. AB - Microbial proliferation and biofilm formation on biologic or inert substrates are characteristics of invasive Staphylococcus aureus infections and is associated with phenotypic alterations such as reduced antimicrobial susceptibility. To identify genes which are typically expressed in biofilms, a micro representational-difference analysis (micro-RDA) was adapted for gram-positive bacteria and used with cDNA derived from populations of S. aureus DSM 20231 growing in a biofilm or plankonically. In comparison to previously described cDNA RDA protocols, micro-RDA has the advantages that only minimal quantities of total RNA are needed and, most importantly, that total RNA can be used since the large amount of rRNA in total RNA does not interfere with the micro-RDA procedure. Using a series of spiked controls with various amounts of MS2 RNA in a background of total RNA from S. aureus, the equivalent of five copies of MS2 per cell were detectable after three rounds of subtractive enrichment. Five genes were identified as being differentially expressed in biofilm versus planktonic cultures. These genes revealed homology to a threonyl-tRNA synthetase, a phosphoglycerate mutase, a triosephosphate isomerase, an alcohol dehydrogenase I, and a ClpC ATPase. Differential levels of expression were subsequently confirmed by standard Northern blotting. In conclusion, micro-RDA is a sensitive and specific method to detect transcripts differentially expressed as a function of different S. aureus growth conditions. PMID- 11425709 TI - Identification of deoxynivalenol- and nivalenol-producing chemotypes of Gibberella zeae by using PCR. AB - Gibberella zeae, a major cause of cereal scab, may be divided into two chemotypes based on production of the trichothecenes deoxynivalenol (DON) and nivalenol (NIV). We cloned and sequenced the gene cluster for trichothecene biosynthesis from each chemotype. G. zeae H-11 is a DON producer isolated from corn, and G. zeae 88-1 is a NIV producer from barley. We sequenced a 23-kb gene cluster from H 11 and a 26-kb cluster from 88-1, along with the unlinked Tri101 genes. Each gene cluster contained 10 Tri gene homologues in the same order and transcriptional directions as those of Fusarium sporotrichioides. Between H-11 and 88-1 all of the Tri homologues except Tri7 were conserved, with identities ranging from 88 to 98% and 82 to 99% at the nucleotide and amino acid levels, respectively. The Tri7 sequences were only 80% identical at the nucleotide level. We aligned the Tri7 genes and found that the Tri7 open reading frame of H-11 carried several mutations and an insertion containing 10 copies of an 11-bp tandem repeat. The Tri7 gene from 88-1 carried neither the repeat nor the mutations. We assayed 100 G. zeae isolates of both chemotypes by PCR amplification with a primer pair derived from the Tri7 gene and could differentiate the chemotypes by polyacrylamide gel electrophoresis. The PCR-based method developed in this study should provide a simple and reliable diagnostic tool for differentiating the two chemotypes of G. zeae. PMID- 11425710 TI - Detoxification of corn antimicrobial compounds as the basis for isolating Fusarium verticillioides and some other Fusarium species from corn. AB - The preformed antimicrobial compounds produced by maize, 2,4-dihydroxy-7-methoxy 2H-1,4-benzoxazin-3-one and its desmethoxy derivative 2,4-dihydroxy-2H-1,4 benzoxazin-3-one, are highly reactive benzoxazinoids that quickly degrade to the antimicrobials 6-methoxy-2-benzoxazolinone (MBOA) and 2-benzoxazolinone (BOA), respectively. Fusarium verticillioides (= F. moniliforme) is highly tolerant to MBOA and BOA and can actively transform these compounds to nontoxic metabolites. Eleven of 29 Fusarium species had some level of tolerance to MBOA and BOA; the most tolerant, in decreasing order, were F. verticillioides, F. subglutinans, F. cerealis (= F. crookwellense), and F. graminearum. The difference in tolerance among species was due to their ability to detoxify the antimicrobials. The limited number of species having tolerance suggested the potential utility of these compounds as biologically active agents for inclusion within a semiselective isolation medium. By replacing the pentachloronitrobenzene in Nash Snyder medium with 1.0 mg of BOA per ml, we developed a medium that resulted in superior frequencies of isolation of F. verticillioides from corn while effectively suppressing competing fungi. Since the BOA medium provided consistent, quantitative results with reduced in vitro and taxonomic efforts, it should prove useful for surveys of F. verticillioides infection in field samples. PMID- 11425711 TI - Quorum sensing in the dimorphic fungus Candida albicans is mediated by farnesol. AB - The inoculum size effect in the dimorphic fungus Candida albicans results from production of an extracellular quorum-sensing molecule (QSM). This molecule prevents mycelial development in both a growth morphology assay and a differentiation assay using three chemically distinct triggers for germ tube formation (GTF): L-proline, N-acetylglucosamine, and serum (either pig or fetal bovine). In all cases, the presence of QSM prevents the yeast-to-mycelium conversion, resulting in actively budding yeasts without influencing cellular growth rates. QSM exhibits general cross-reactivity within C. albicans in that supernatants from strain A72 are active on five other strains of C. albicans and vice versa. The QSM excreted by C. albicans is farnesol (C(15)H(26)O; molecular weight, 222.37). QSM is extracellular, and is produced continuously during growth and over a temperature range from 23 to 43 degrees C, in amounts roughly proportional to the CFU/milliliter. Production is not dependent on the type of carbon source nor nitrogen source or on the chemical nature of the growth medium. Both commercial mixed isomer and (E,E)-farnesol exhibited QSM activity (the ability to prevent GTF) at a level sufficient to account for all the QSM activity present in C. albicans supernatants, i.e., 50% GTF at ca. 30 to 35 microM. Nerolidol was ca. two times less active than farnesol. Neither geraniol (C(10)), geranylgeraniol (C(20)), nor farnesyl pyrophosphate had any QSM activity. PMID- 11425712 TI - Chlorine dioxide inactivation of Cryptosporidium parvum oocysts and bacterial spore indicators. AB - Cryptosporidium parvum, which is resistant to chlorine concentrations typically used in water treatment, is recognized as a significant waterborne pathogen. Recent studies have demonstrated that chlorine dioxide is a more efficient disinfectant than free chlorine against Cryptosporidium oocysts. It is not known, however, if oocysts from different suppliers are equally sensitive to chlorine dioxide. This study used both a most-probable-number-cell culture infectivity assay and in vitro excystation to evaluate chlorine dioxide inactivation kinetics in laboratory water at pH 8 and 21 degrees C. The two viability methods produced significantly different results (P < 0.05). Products of disinfectant concentration and contact time (Ct values) of 1,000 mg. min/liter were needed to inactivate approximately 0.5 log(10) and 2.0 log(10) units (99% inactivation) of C. parvum as measured by in vitro excystation and cell infectivity, respectively, suggesting that excystation is not an adequate viability assay. Purified oocysts originating from three different suppliers were evaluated and showed marked differences with respect to their resistance to inactivation when using chlorine dioxide. Ct values of 75, 550, and 1,000 mg. min/liter were required to achieve approximately 2.0 log(10) units of inactivation with oocysts from different sources. Finally, the study compared the relationship between easily measured indicators, including Bacillus subtilis (aerobic) spores and Clostridium sporogenes (anaerobic) spores, and C. parvum oocysts. The bacterial spores were found to be more sensitive to chlorine dioxide than C. parvum oocysts and therefore could not be used as direct indicators of C. parvum inactivation for this disinfectant. In conclusion, it is suggested that future studies address issues such as oocyst purification protocols and the genetic diversity of C. parvum, since these factors might affect oocyst disinfection sensitivity. PMID- 11425713 TI - Ecological physiology of Synechococcus sp. strain SH-94-5, a naturally occurring cyanobacterium deficient in nitrate assimilation. AB - Synechococcus sp. strain SH-94-5 is a nitrate assimilation-deficient cyanobacterium which was isolated from an ammonium-replete hot spring in central Oregon. While this clone could grow on ammonium and some forms of organic nitrogen as sole nitrogen sources, it could not grow on either nitrate or nitrite, even under conditions favoring passive diffusion. It was determined that this clone does not express functional nitrate reductase or nitrite reductase and that the lack of activity of either enzyme is not due to inactivation of the cyanobacterial nitrogen control protein NtcA. A few other naturally occurring cyanobacterial strains are also nitrate assimilation deficient, and phylogenetic analyses indicated that the ability to utilize nitrate has been independently lost at least four times during the evolutionary history of the cyanobacteria. This phenotype is associated with the presence of environmental ammonium, a negative regulator of nitrate assimilation gene expression, which may indicate that natural selection to maintain functional copies of nitrate assimilation genes has been relaxed in these habitats. These results suggest how the evolutionary fates of conditionally expressed genes might differ between environments and thereby effect ecological divergence and biogeographical structure in the microbial world. PMID- 11425714 TI - Expression of the p20 gene from Bacillus thuringiensis H-14 increases Cry11A toxin production and enhances mosquito-larvicidal activity in recombinant gram negative bacteria. AB - Experimental analyses with recombinant Escherichia coli and Pseudomonas putida transformed with plasmids bearing genes coding for the Cry11A toxin and P20 protein from Bacillus thuringiensis H-14 showed that cells producing both proteins were more toxic when fed to third-instar Aedes aegypti larvae than were cells expressing cry11A alone; the 50% lethal concentrations were in the range of 10(4) to 10(5) cells/ml. Western blots revealed a higher production of Cry11A when the p20 gene was coexpressed. Cry11A was detected primarily in insoluble form in recombinant cells. Cry11A was not detected in P. putida when P20 was not coproduced, and these recombinants were not toxic to larvae, whereas P. putida recombinants producing both proteins were toxic at concentrations similar to those for E. coli. A coelution experiment was conducted, in which a p20 gene construct producing the P20 protein with an extension of six histidines on the C terminus was mixed with the Cry11A protein. The results showed that Cry11A bound to the P20(His(6)) on a nickel chelating column, whereas Cry11A produced without the P20(His(6)) protein was washed through the column, thus indicating that Cry11A and P20 physically interact. Thus, P20 protein either stabilizes Cry11A or helps it attain the folding important for its toxic activity. PMID- 11425715 TI - Detection and identification of mammalian reoviruses in surface water by combined cell culture and reverse transcription-PCR. AB - Reoviruses are a common class of enteric viruses capable of infecting a broad range of mammalian species, typically with low pathogenicity. Previous studies have shown that reoviruses are common in raw water sources and are often found along with other animal viruses. This suggests that in addition to the commonly monitored enteroviruses, reoviruses might serve as an informative target for monitoring fecal contamination of drinking water sources. Mammalian reoviruses were detected and identified by a combined cell culture-reverse transcription-PCR (RT-PCR) assay with novel primers targeting the L3 gene that encodes the lambda3 major core protein. Five of 26 (19.2%) cytopathic effect-positive cell culture lysates inoculated with surface water were positive for reoviruses by RT-PCR. DNA sequence analysis of RT-PCR products revealed significant sequence diversity among isolates, which is consistent with the sequence diversity among previously characterized mammalian reoviruses. Sequence analysis revealed persistence of a reovirus genotype at a single sampling site, while a sample from another site contained two different reovirus genotypes. PMID- 11425716 TI - Use of long-range repetitive element polymorphism-PCR to differentiate Bacillus anthracis strains. AB - The genome of Bacillus anthracis is extremely monomorphic, and thus individual strains have often proven to be recalcitrant to differentiation at the molecular level. Long-range repetitive element polymorphism-PCR (LR REP-PCR) was used to differentiate various B. anthracis strains. A single PCR primer derived from a repetitive DNA element was able to amplify variable segments of a bacterial genome as large as 10 kb. We were able to characterize five genetically distinct groups by examining 105 B. anthracis strains of diverse geographical origins. All B. anthracis strains produced fingerprints comprising seven to eight bands, referred to as "skeleton" bands, while one to three "diagnostic" bands differentiated between B. anthracis strains. LR REP-PCR fingerprints of B. anthracis strains showed very little in common with those of other closely related species such as B. cereus, B. thuringiensis, and B. mycoides, suggesting relative heterogeneity among the non-B. anthracis strains. Fingerprints from transitional non-B. anthracis strains, which possessed the B. anthracis chromosomal marker Ba813, scarcely resembled those observed for any of the five distinct B. anthracis groups that we have identified. The LR REP-PCR method described in this report provides a simple means of differentiating B. anthracis strains. PMID- 11425717 TI - Low-pressure UV inactivation and DNA repair potential of Cryptosporidium parvum oocysts. AB - Because Cryptosporidium parvum oocysts are very resistant to conventional water treatment processes, including chemical disinfection, we determined the kinetics and extent of their inactivation by monochromatic, low-pressure (LP), mercury vapor lamp UV radiation and their subsequent potential for DNA repair of UV damage. A UV collimated-beam apparatus was used to expose suspensions of purified C. parvum oocysts in phosphate-buffered saline, pH 7.3, at 25 degrees C to various doses of monochromatic LP UV. C. parvum infectivity reductions were rapid, approximately first order, and at a dose of 3 mJ/cm(2) (=30 J/m(2)), the reduction reached the cell culture assay detection limit of approximately 3 log(10). At UV doses of 1.2 and 3 mJ/cm(2), the log(10) reductions of C. parvum oocyst infectivity were not significantly different for control oocysts and those exposed to dark or light repair conditions for UV-induced DNA damage. These results indicate that C. parvum oocysts are very sensitive to inactivation by low doses of monochromatic LP UV radiation and that there is no phenotypic evidence of either light or dark repair of UV-induced DNA damage. PMID- 11425718 TI - Functional analysis of the Lactococcus lactis galU and galE genes and their impact on sugar nucleotide and exopolysaccharide biosynthesis. AB - We studied the UDP-glucose pyrophosphorylase (galU) and UDP-galactose epimerase (galE) genes of Lactococcus lactis MG1363 to investigate their involvement in biosynthesis of UDP-glucose and UDP-galactose, which are precursors of glucose- and galactose-containing exopolysaccharides (EPS) in L. lactis. The lactococcal galU gene was identified by a PCR approach using degenerate primers and was found by Northern blot analysis to be transcribed in a monocistronic RNA. The L. lactis galU gene could complement an Escherichia coli galU mutant, and overexpression of this gene in L. lactis under control of the inducible nisA promoter resulted in a 20-fold increase in GalU activity. Remarkably, this resulted in approximately eightfold increases in the levels of both UDP-glucose and UDP-galactose. This indicated that the endogenous GalE activity is not limiting and that the GalU activity level in wild-type cells controls the biosynthesis of intracellular UDP glucose and UDP-galactose. The increased GalU activity did not significantly increase NIZO B40 EPS production. Disruption of the galE gene resulted in poor growth, undetectable intracellular levels of UDP-galactose, and elimination of EPS production in strain NIZO B40 when cells were grown in media with glucose as the sole carbon source. Addition of galactose restored wild-type growth in the galE disruption mutant, while the level of EPS production was approximately one half the wild-type level. PMID- 11425719 TI - Characterization of a heme-dependent catalase from Methanobrevibacter arboriphilus. AB - Recently it was reported that methanogens of the genus Methanobrevibacter exhibit catalase activity. This was surprising, since Methanobrevibacter species belong to the order Methanobacteriales, which are known not to contain cytochromes and to lack the ability to synthesize heme. We report here that Methanobrevibacter arboriphilus strains AZ and DH1 contained catalase activity only when the growth medium was supplemented with hemin. The heme catalase was purified and characterized, and the encoding gene was cloned. The amino acid sequence of the catalase from the methanogens is most similar to that of Methanosarcina barkeri. PMID- 11425720 TI - Effects of rice seed surface sterilization with hypochlorite on inoculated Burkholderia vietnamiensis. AB - When a combination of hydrogen peroxide and hypochlorite was used to surface sterilize rice seeds, a 10(2)- to 10(4)-fold decrease in CFU was observed during the first 15 h after inoculation of the rice rhizosphere organism Burkholderia vietnamiensis TVV75. This artifact could not be eliminated simply by rinsing the seeds, even thoroughly, with sterile distilled water. When growth resumed, a significant increase in the frequency of rifampin- and nalidixic acid-resistant mutants in the population was observed compared to the control without seeds. This phenomenon was a specific effect of hypochlorite; it was not observed with hydrogen peroxide alone. It was also not observed when the effect of hypochlorite was counteracted by sodium thiosulfate. We hypothesized that the hypochlorite used for disinfection reacted with the rice seed surface, forming a chlorine cover which was not removed by rinsing and generated mutagenic chloramines. We studied a set of rifampin- and nalidixic acid-resistant mutants obtained after seed surface sterilization. The corresponding rpoB and gyrA genes were amplified and sequenced to characterize the induced mutations. The mutations in five of seven nalidixic acid-resistant mutants and all of the rifampin-resistant mutants studied were found to correspond to single amino acid substitutions. Hypochlorite surface sterilization can thus be a source of artifacts when the initial bacterial colonization of a plant is studied. PMID- 11425721 TI - Cattle water troughs as reservoirs of Escherichia coli O157. AB - Environmental survival of Escherichia coli O157 may play an important role in the persistence and dissemination of this organism on farms. The survival of culturable and infectious E. coli O157 was studied using microcosms simulating cattle water troughs. Culturable E. coli O157 survived for at least 245 days in the microcosm sediments. Furthermore, E. coli O157 strains surviving more than 6 months in contaminated microcosms were infectious to a group of 10-week-old calves. Fecal excretion of E. coli O157 by these calves persisted for 87 days after challenge. Water trough sediments contaminated with feces from cattle excreting E. coli O157 may serve as a long-term reservoir of this organism on farms and a source of infection for cattle. PMID- 11425722 TI - Killer toxin of Kluyveromyces phaffii DBVPG 6076 as a biopreservative agent to control apiculate wine Yeasts. AB - The use of Kluyveromyces phaffii DBVPG 6076 killer toxin against apiculate wine yeasts has been investigated. The killer toxin of K. phaffii DBVPG 6076 showed extensive anti-Hanseniaspora activity against strains isolated from grape samples. The proteinaceous killer toxin was found to be active in the pH range of 3 to 5 and at temperatures lower than 40 degrees C. These biochemical properties would allow the use of K. phaffii killer toxin in wine making. Fungicidal or fungistatic effects depend on the toxin concentration. Toxin concentrations present in the supernatant during optimal conditions of production (14.3 arbitrary units) exerted a fungicidal effect on a sensitive strain of Hanseniaspora uvarum. At subcritical concentrations (fungistatic effect) the saturation kinetics observed with the increased ratio of killer toxin to H. uvarum cells suggest the presence of a toxin receptor. The inhibitory activity exerted by the killer toxin present in grape juice was comparable to that of sulfur dioxide. The findings presented suggest that the K. phaffii DBVPG 6076 killer toxin has potential as a biopreservative agent in wine making. PMID- 11425723 TI - Overproduction of L-Lysine from methanol by Methylobacillus glycogenes derivatives carrying a plasmid with a mutated dapA gene. AB - The dapA gene, encoding dihydrodipicolinate synthase (DDPS) partially desensitized to inhibition by L-lysine, was cloned from an L-threonine- and L lysine-coproducing mutant of the obligate methylotroph Methylobacillus glycogenes DHL122 by complementation of the nutritional requirement of an Escherichia coli dapA mutant. Introduction of the dapA gene into DHL122 and AL119, which is the parent of DHL122 and an L-threonine producing mutant, elevated the specific activity of DDPS 20-fold and L-lysine production 2- to 3-fold with concomitant reduction of L-threonine in test tube cultures. AL119 containing the dapA gene produced 8 g of L-lysine per liter in a 5-liter jar fermentor from methanol as a substrate. Analysis of the nucleotide sequence of the dapA gene shows that it encodes a peptide with an M(r) of 30,664 and that the encoded amino acid sequence is extensively homologous to those of other organisms. In order to study the mutation that occurred in DHL122, the dapA genes of the wild type and AL119 were cloned and sequenced. Comparison of the nucleotide sequences of the dapA genes revealed that the amino acid at residue 88 was F in DHL122 whereas it was L in the wild type and AL119, suggesting that this amino acid alteration that occurred in DHL122 caused the partial desensitization of DDPS to the inhibition by L lysine. The similarity in the amino acid sequences of DDPS in M. glycogenes and other organisms suggests that the mutation of the dapA gene in DHL122 is located in the region concerned with interaction of the allosteric effector, L-lysine. PMID- 11425724 TI - Effect of fermented feed on the microbial population of the gastrointestinal tracts of pigs. AB - An in vivo experiment was performed with pigs to study the inhibitory effect of fermented feed on the bacterial population of the gastrointestinal tract. Results demonstrated a significant positive correlation between pH and lactobacilli in the stomach contents of pigs in dry feed as well as in the stomach contents of pigs fed fermented feed. Furthermore, a significant positive correlation between the pH and the numbers of bacteria in the family Enterobacteriaceae in the contents of the stomach of pigs fed dry feed was found. In the stomach contents of pigs fed fermented feed, a significant negative correlation was found between the concentration of the undissociated form of lactic acid and the numbers of Enterobacteriaceae. The numbers of Enterobacteriaceae in the contents of the stomach, ileum, cecum, colon, and rectum of pigs fed fermented feed were significantly lower compared with the contents of the stomach, ileum, caecum, colon, and rectum of pigs fed dry feed. The numbers of total lactobacilli were significantly higher in the stomach contents of pigs fed fermented feed and in the ileum contents of one pig group fed fermented feed compared with the contents of pigs fed dry feed. However, the influence of lactobacilli on numbers of Enterobacteriaceae could not be demonstrated. It was concluded that fermented feed influences the bacterial ecology of the gastrointestinal tract and reduces the levels of Enterobacteriaceae in the different parts of the gastrointestinal tract. PMID- 11425725 TI - Microbial diversity of the brine-seawater interface of the Kebrit Deep, Red Sea, studied via 16S rRNA gene sequences and cultivation methods. AB - The brine-seawater interface of the Kebrit Deep, northern Red Sea, was investigated for the presence of microorganisms using phylogenetic analysis combined with cultivation methods. Under strictly anaerobic culture conditions, novel halophiles were isolated. The new rod-shaped isolates belong to the halophilic genus Halanaerobium and are the first representatives of the genus obtained from deep-sea, anaerobic brine pools. Within the genus Halanaerobium, they represent new species which grow chemoorganotrophically at NaCl concentrations ranging from 5 to 34%. The cellular fatty acid compositions are consistent with those of other Halanaerobium representatives, showing unusually large amounts of Delta7 and Delta11 16:1 fatty acids. Phylogenetic analysis of the brine-seawater interface sample revealed the presence of various bacterial 16S rRNA gene sequences dominated by cultivated members of the bacterial domain, with the majority affiliated with the genus Halanaerobium. The new Halanaerobium 16S rRNA clone sequences showed the highest similarity (99.9%) to the sequence of isolate KT-8-13 from the Kebrit Deep brine. In this initial survey, our polyphasic approach demonstrates that novel halophiles thrive in the anaerobic, deep-sea brine pool of the Kebrit Deep, Red Sea. They may contribute significantly to the anaerobic degradation of organic matter enriched at the brine-seawater interface. PMID- 11425726 TI - Surface binding of aflatoxin B(1) by lactic acid bacteria. AB - Specific lactic acid bacterial strains remove toxins from liquid media by physical binding. The stability of the aflatoxin B(1) complexes formed with 12 bacterial strains in both viable and nonviable (heat- or acid-treated) forms was assessed by repetitive aqueous extraction. By the fifth extraction, up to 71% of the total aflatoxin B(1) remained bound. Nonviable bacteria retained the highest amount of aflatoxin B(1). Lactobacillus rhamnosus strain GG (ATCC 53103) and L. rhamnosus strain LC-705 (DSM 7061) removed aflatoxin B(1) from solution most efficiently and were selected for further study. The accessibility of bound aflatoxin B(1) to an antibody in an indirect competitive inhibition enzyme-linked immunosorbent assay suggests that surface components of these bacteria are involved in binding. Further evidence is the recovery of around 90% of the bound aflatoxin from the bacteria by solvent extraction. Autoclaving and sonication did not release any detectable aflatoxin B(1). Variation in temperature (4 to 37 degrees C) and pH (2 to 10) did not have any significant effect on the amount of aflatoxin B(1) released. Binding of aflatoxin B(1) appears to be predominantly extracellular for viable and heat-treated bacteria. Acid treatment may permit intracellular binding. In all cases, binding is of a reversible nature, but the stability of the complexes formed depends on strain, treatment, and environmental conditions. PMID- 11425727 TI - Membrane permeabilization in relation to inactivation kinetics of Lactobacillus species due to pulsed electric fields. AB - Membrane permeabilization due to pulsed electric field (PEF) treatment of gram positive Lactobacillus cells was investigated by using propidium iodide uptake and single-cell analysis with flow cytometry. Electric field strength, energy input, treatment time, and growth phase affected membrane permeabilization of Lactobacillus plantarum during PEF treatment. A correlation between PEF inactivation and membrane permeabilization of L. plantarum cells was demonstrated, whereas no relationship was observed between membrane permeabilization and heat inactivation. The same results were obtained with a Lactobacillus fermentum strain, but the latter organism was more PEF resistant and exhibited less membrane permeabilization, indicating that various bacteria have different responses to PEF treatment. While membrane permeabilization was the main factor involved in the mechanism of inactivation, the growth phase and the acidity of the environment also influenced inactivation. By using flow cytometry it was possible to sort cells in the L. plantarum population based on different cell sizes and shapes, and the results were confirmed by image analysis. An apparent effect of morphology on membrane permeabilization was observed, and larger cells were more easily permeabilized than smaller cells. In conclusion, our results indicate that the ability of PEF treatment to cause membrane permeabilization is an important factor in determining inactivation. This finding should have an effect on the final choice of the processing parameters used so that all microorganisms can be inactivated and, consequently, on the use of PEF treatment as an alternative method for preserving food products. PMID- 11425728 TI - Role of fatty acid de novo biosynthesis in polyhydroxyalkanoic acid (PHA) and rhamnolipid synthesis by pseudomonads: establishment of the transacylase (PhaG) mediated pathway for PHA biosynthesis in Escherichia coli. AB - Since Pseudomonas aeruginosa is capable of biosynthesis of polyhydroxyalkanoic acid (PHA) and rhamnolipids, which contain lipid moieties that are derived from fatty acid biosynthesis, we investigated various fab mutants from P. aeruginosa with respect to biosynthesis of PHAs and rhamnolipids. All isogenic fabA, fabB, fabI, rhlG, and phaG mutants from P. aeruginosa showed decreased PHA accumulation and rhamnolipid production. In the phaG (encoding transacylase) mutant rhamnolipid production was only slightly decreased. Expression of phaG from Pseudomonas putida and expression of the beta-ketoacyl reductase gene rhlG from P. aeruginosa in these mutants indicated that PhaG catalyzes diversion of intermediates of fatty acid de novo biosynthesis towards PHA biosynthesis, whereas RhlG catalyzes diversion towards rhamnolipid biosynthesis. These data suggested that both biosynthesis pathways are competitive. In order to investigate whether PhaG is the only linking enzyme between fatty acid de novo biosynthesis and PHA biosynthesis, we generated five Tn5 mutants of P. putida strongly impaired in PHA production from gluconate. All mutants were complemented by the phaG gene from P. putida, indicating that the transacylase-mediated PHA biosynthesis route represents the only metabolic link between fatty acid de novo biosynthesis and PHA biosynthesis in this bacterium. The transacylase-mediated PHA biosynthesis route from gluconate was established in recombinant E. coli, coexpressing the class II PHA synthase gene phaC1 together with the phaG gene from P. putida, only when fatty acid de novo biosynthesis was partially inhibited by triclosan. The accumulated PHA contributed to 2 to 3% of cellular dry weight. PMID- 11425729 TI - Glutamate decarboxylase genes as a prescreening marker for detection of pathogenic Escherichia coli groups. AB - The enzyme glutamate decarboxylase (GAD) is prevalent in Escherichia coli but few strains in the various pathogenic E. coli groups have been tested for GAD. Using PCR primers that amplify a 670-bp segment from the gadA and gadB genes encoding GAD, we examined the distribution of the gadAB genes among enteric bacteria. Analysis of 173 pathogenic E. coli strains, including 125 enterohemorrhagic E. coli isolates of the O157:H7 serotype and its phenotypic variants and 48 isolates of enteropathogenic E. coli, enterotoxigenic E. coli, enteroinvasive E. coli, and other Shiga toxin-producing E. coli (STEC) serotypes, showed that gadAB genes were present in all these strains. Among the 22 non-E. coli isolates tested, only the 6 Shigella spp. carried gadAB. Analysis of naturally contaminated water and food samples using a gadAB-specific DNA probe that was labeled with digoxigenin showed that a gadAB-based assay is as reliable as standard methods that enumerate E. coli organisms on the basis of lactose fermentation. The presence of few E. coli cells initially seeded into produce rinsates could be detected by PCR to gadA/B genes after overnight enrichment. A multiplex PCR assay using the gadAB primers in combination with primers to Shiga toxin (Stx) genes stx(1) and stx(2) was effective in detecting STEC from the enrichment medium after seeding produce rinsate samples with as few as 2 CFU. The gadAB primers may be multiplexed with primers to other trait virulence markers to specifically identify other pathogenic E. coli groups. PMID- 11425730 TI - Comparison of genotypes and serotypes of Campylobacter jejuni isolated from Danish wild mammals and birds and from broiler flocks and humans. AB - The incidence of human infection with Campylobacter jejuni is increasing in most developed countries and the reason for this is largely unknown. Although poultry meat is considered to be a major source, it is evident that other reservoirs exist, possibly common to humans and poultry. Environmental sources are believed to be important reservoirs of Campylobacter infection in broiler chicken flocks. We investigated the potential importance of wildlife as a source of infection in commercial poultry flocks and in humans by comparing the serotype distributions, fla types, and macrorestriction profiles (MRPs) of C. jejuni isolates from different sources. The serotype distribution in wildlife was significantly different from the known distributions in broilers and humans. Considerable sero- and genotype diversity was found within the wildlife collection, although two major groups of isolates within serotype O:12 and the O:4 complex were found. Common clonal lines among wildlife, chicken, and/or human isolates were identified within serotype O:2 and the O:4 complex. However, MRPs of O:12 and O:38 strains isolated from wildlife and other sources indicated that some clonal lines propagated in a wide selection of animal species but were not detected in humans or broilers in this study. The applied typing methods successfully identified different clonal groups within a strain collection showing large genomic diversity. However, the relatively low number of wildlife strains with an inferred clonal relationship to human and chicken strains suggests that the importance of wildlife as a reservoir of infection is limited. PMID- 11425731 TI - Comparison of different approaches to quantify Staphylococcus aureus cells by real-time quantitative PCR and application of this technique for examination of cheese. AB - Two different real-time quantitative PCR (RTQ-PCR) approaches were applied for PCR-based quantification of Staphylococcus aureus cells by targeting the thermonuclease (nuc) gene. Purified DNA extracts from pure cultures of S. aureus were quantified in a LightCycler system using SYBR Green I. Quantification proved to be less sensitive (60 nuc gene copies/microl) than using a fluorigenic TaqMan probe (6 nuc gene copies/microl). Comparison of the LightCycler system and the well-established ABI Prism 7700 SDS with TaqMan probes revealed no statistically significant differences with respect to sensitivity and reproducibility. Application of the RTQ-PCR assay to quantify S. aureus cells in artificially contaminated cheeses of different types achieved sensitivities from 1.5 x 10(2) to 6.4 x 10(2) copies of the nuc gene/2 g, depending on the cheese matrix. The coefficients of correlation between log CFU and nuc gene copy numbers ranged from 0.979 to 0.998, thus enabling calculation of the number of CFU of S. aureus in cheese by performing RTQ-PCR. PMID- 11425732 TI - Bioremediation (natural attenuation and biostimulation) of diesel-oil contaminated soil in an alpine glacier skiing area. AB - We investigated the feasibility of bioremediation as a treatment option for a chronically diesel-oil-polluted soil in an alpine glacier area at an altitude of 2,875 m above sea level. To examine the efficiencies of natural attenuation and biostimulation, we used field-incubated lysimeters (mesocosms) with unfertilized and fertilized (N-P-K) soil. For three summer seasons (July 1997 to September 1999), we monitored changes in hydrocarbon concentrations in soil and soil leachate and the accompanying changes in soil microbial counts and activity. A significant reduction in the diesel oil level could be achieved. At the end of the third summer season (after 780 days), the initial level of contamination (2,612 +/- 70 microg of hydrocarbons g [dry weight] of soil(-1)) was reduced by (50 +/- 4)% and (70 +/- 2)% in the unfertilized and fertilized soil, respectively. Nonetheless, the residual levels of contamination (1,296 +/- 110 and 774 +/- 52 microg of hydrocarbons g [dry weight] of soil(-1) in the unfertilized and fertilized soil, respectively) were still high. Most of the hydrocarbon loss occurred during the first summer season ([42 +/- 6]% loss) in the fertilized soil and during the second summer season ([41 +/- 4]% loss) in the unfertilized soil. In the fertilized soil, all biological parameters (microbial numbers, soil respiration, catalase and lipase activities) were significantly enhanced and correlated significantly with each other, as well as with the residual hydrocarbon concentration, pointing to the importance of biodegradation. The effect of biostimulation of the indigenous soil microorganisms declined with time. The microbial activities in the unfertilized soil fluctuated around background levels during the whole study. PMID- 11425733 TI - Nuclear magnetic resonance analysis of [1-13C]dimethylsulfoniopropionate (DMSP) and [1-13C]acrylate metabolism by a DMSP lyase-producing marine isolate of the alpha-subclass of Proteobacteria. AB - The prominence of the alpha-subclass of Proteobacteria in the marine bacterioplankton community and their role in dimethylsulfide (DMS) production has prompted a detailed examination of dimethylsulfoniopropionate (DMSP) metabolism in a representative isolate of this phylotype, strain LFR. [1-(13)C]DMSP was synthesized, and its metabolism and that of its cleavage product, [1 (13)C]acrylate, were studied using nuclear magnetic resonance (NMR) spectroscopy. [1-(13)C]DMSP additions resulted in the intracellular accumulation and then disappearance of both [1-(13)C]DMSP and [1-(13)C]beta-hydroxypropionate ([1 (13)C]beta-HP), a degradation product. Acrylate, the immediate product of DMSP cleavage, apparently did not accumulate to high enough levels to be detected, suggesting that it was rapidly beta-hydroxylated upon formation. When [1 (13)C]acrylate was added to cell suspensions of strain LFR it was metabolized to [1-(13)C]beta-HP extracellularly, where it first accumulated and was then taken up in the cytosol where it subsequently disappeared, indicating that it was directly decarboxylated. These results were interpreted to mean that DMSP was taken up and metabolized by an intracellular DMSP lyase and acrylase, while added acrylate was beta-hydroxylated on (or near) the cell surface to beta-HP, which accumulated briefly and was then taken up by cells. Growth on acrylate (versus that on glucose) stimulated the rate of acrylate metabolism eightfold, indicating that it acted as an inducer of acrylase activity. DMSP, acrylate, and beta-HP all induced DMSP lyase activity. A putative model is presented that best fits the experimental data regarding the pathway of DMSP and acrylate metabolism in the alpha-proteobacterium, strain LFR. PMID- 11425734 TI - Natural transformation in mesophilic and thermophilic bacteria: identification and characterization of novel, closely related competence genes in Acinetobacter sp. strain BD413 and Thermus thermophilus HB27. AB - The mesophile Acinetobacter sp. strain BD413 and the extreme thermophile Thermus thermophilus HB27 display high frequencies of natural transformation. In this study we identified and characterized a novel competence gene in Acinetobacter sp. strain BD413, comA, whose product displays significant similarities to the competence proteins ComA and ComEC in Neisseria and Bacillus species. Transcription of comA correlated with growth phase-dependent transcriptional regulation of the recently identified pilin-like factors of the transformation machinery. This finding strongly suggests that comA is part of a competence regulon. Examination of the genome sequence of T. thermophilus HB27 led to detection of a comA/comEC-like open reading frame (ORF) which is flanked by an ORF whose product shows significant similarities to the Bacillus subtilis competence protein ComEA. To examine whether these two ORFs, designated comEC and comEA, are implicated in natural transformation of T. thermophilus HB27, both were disrupted by using a thermostable kanamycin resistance marker. Natural transformation in comEC mutants was reduced 1,000-fold, whereas in comEA mutants the natural transformation phenotype was completely eliminated. These results strongly suggest that both genes, comEC and comEA, are required for natural transformation in T. thermophilus HB27. Several transmembrane alpha-helices are predicted based on the amino acid sequences of ComA in Acinetobacter sp. strain BD413 and ComEC in T. thermophilus HB27, which suggests that ComA and ComEC are located in the inner membrane and function in DNA transport through the cytoplasmic membrane. PMID- 11425735 TI - Diversity and characterization of sulfate-reducing bacteria in groundwater at a uranium mill tailings site. AB - Microbially mediated reduction and immobilization of U(VI) to U(IV) plays a role in both natural attenuation and accelerated bioremediation of uranium contaminated sites. To realize bioremediation potential and accurately predict natural attenuation, it is important to first understand the microbial diversity of such sites. In this paper, the distribution of sulfate-reducing bacteria (SRB) in contaminated groundwater associated with a uranium mill tailings disposal site at Shiprock, N.Mex., was investigated. Two culture-independent analyses were employed: sequencing of clone libraries of PCR-amplified dissimilatory sulfite reductase (DSR) gene fragments and phospholipid fatty acid (PLFA) biomarker analysis. A remarkable diversity among the DSR sequences was revealed, including sequences from delta-Proteobacteria, gram-positive organisms, and the Nitrospira division. PLFA analysis detected at least 52 different mid-chain-branched saturate PLFA and included a high proportion of 10me16:0. Desulfotomaculum and Desulfotomaculum-like sequences were the most dominant DSR genes detected. Those belonging to SRB within delta-Proteobacteria were mainly recovered from low uranium (< or =302 ppb) samples. One Desulfotomaculum-like sequence cluster overwhelmingly dominated high-U (>1,500 ppb) sites. Logistic regression showed a significant influence of uranium concentration over the dominance of this cluster of sequences (P = 0.0001). This strong association indicates that Desulfotomaculum has remarkable tolerance and adaptation to high levels of uranium and suggests the organism's possible involvement in natural attenuation of uranium. The in situ activity level of Desulfotomaculum in uranium contaminated environments and its comparison to the activities of other SRB and other functional groups should be an important area for future research. PMID- 11425736 TI - Duplication of hemolysin genes in a virulent isolate of Vibrio harveyi. AB - Vibrio harveyi VIB 645, which is very pathogenic towards salmonids and produces extracellular product with a high titer of hemolytic activity towards fish erythrocytes, was found to contain two closely related hemolysin genes (designated vhhA and vhhB), whereas the majority of strains examined (11 of 13) carried only a single hemolysin gene. Both genes from VIB 645 were cloned and sequenced. The open reading frames (ORFs) of vhhA and vhhB shared a high level of identity (98.8%) and were predicted to encode identical polypeptides comprising 418 amino acid residues. The VHH protein shows homology to the lecithinase of V. mimicus and V. cholerae. Transformants of Escherichia coli containing the ORF of either vhhA or vhhB displayed weak hemolytic activity in rainbow trout blood agar. The hemolytic activity was very high when the ORF of vhhB was cloned in E. coli together with the native promoter. Surprisingly, the level of vhh-specific RNA transcript produced by VIB 645 was found to be very low. We conclude that the hemolytic phenotype of VIB 645 is not due to increased expression of one or both copies of the vhh gene. PMID- 11425737 TI - Rapid and simple method for the most-probable-number estimation of arsenic reducing bacteria. AB - A rapid and simple most-probable-number (MPN) procedure for the enumeration of dissimilatory arsenic-reducing bacteria (DARB) is presented. The method is based on the specific detection of arsenite, the end product of anaerobic arsenate respiration, by a precipitation reaction with sulfide. After 4 weeks of incubation, the medium for the MPN method is acidified to pH 6 and sulfide is added to a final concentration of about 1 mM. The brightly yellow arsenic trisulfide precipitates immediately and can easily be scored at arsenite concentrations as low as 0.05 mM. Abiotic reduction of arsenate upon sulfide addition, which could yield false positives, apparently produces a soluble As-S intermediate, which does not precipitate until about 1 h after sulfide addition. Using the new MPN method, population estimates of pure cultures of DARB were similar to direct cell counts. MPNs of environmental water and sediment samples yielded DARB numbers between 10(1) and 10(5) cells per ml or gram (dry weight), respectively. Poisoned and sterilized controls showed that potential abiotic reductants in environmental samples did not interfere with the MPN estimates. A major advantage is that the assay can be easily scaled to a microtiter plate format, enabling analysis of large numbers of samples by use of multichannel pipettors. Overall, the MPN method provides a rapid and simple means for estimating population sizes of DARB, a diverse group of organisms for which no comprehensive molecular markers have been developed yet. PMID- 11425738 TI - Reaction of acylated homoserine lactone bacterial signaling molecules with oxidized halogen antimicrobials. AB - Oxidized halogen antimicrobials, such as hypochlorous and hypobromous acids, have been used extensively for microbial control in industrial systems. Recent discoveries have shown that acylated homoserine lactone cell-to-cell signaling molecules are important for biofilm formation in Pseudomonas aeruginosa, suggesting that biofouling can be controlled by interfering with bacterial cell to-cell communication. This study was conducted to investigate the potential for oxidized halogens to react with acylated homoserine lactone-based signaling molecules. Acylated homoserine lactones containing a 3-oxo group were found to rapidly react with oxidized halogens, while acylated homoserine lactones lacking the 3-oxo functionality did not react. The Chromobacterium violaceum CV026 bioassay was used to determine the effects of such reactions on acylated homoserine lactone activity. The results demonstrated that 3-oxo acyl homoserine lactone activity was rapidly lost upon exposure to oxidized halogens; however, acylated homoserine lactones lacking the 3-oxo group retained activity. Experiments with the marine alga Laminaria digitata demonstrated that natural haloperoxidase systems are capable of mediating the deactivation of acylated homoserine lactones. This may illustrate a natural defense mechanism to prevent biofouling on the surface of this marine alga. The Chromobacterium violaceum activity assay illustrates that reactions between 3-oxo acylated homoserine lactone molecules and oxidized halogens do occur despite the presence of biofilm components at much greater concentrations. This work suggests that oxidized halogens may control biofilm not only via a cidal mechanism, but also by possibly interfering with 3-oxo acylated homoserine lactone-based cell signaling. PMID- 11425739 TI - Development of a genetic system for Geobacter sulfurreducens. AB - Members of the genus Geobacter are the dominant metal-reducing microorganisms in a variety of anaerobic subsurface environments and have been shown to be involved in the bioremediation of both organic and metal contaminants. To facilitate the study of the physiology of these organisms, a genetic system was developed for Geobacter sulfurreducens. The antibiotic sensitivity of this organism was characterized, and optimal conditions for plating it at high efficiency were established. A protocol for the introduction of foreign DNA into G. sulfurreducens by electroporation was also developed. Two classes of broad-host range vectors, IncQ and pBBR1, were found to be capable of replication in G. sulfurreducens. In particular, the IncQ plasmid pCD342 was found to be a suitable expression vector for this organism. When the information and novel methods described above were utilized, the nifD gene of G. sulfurreducens was disrupted by the single-step gene replacement method. Insertional mutagenesis of this key gene in the nitrogen fixation pathway impaired the ability of G. sulfurreducens to grow in medium lacking a source of fixed nitrogen. Expression of the nifD gene in trans complemented this phenotype. This paper constitutes the first report of genetic manipulation of a member of the Geobacter genus. PMID- 11425740 TI - Adhesion of Pseudomonas fluorescens (ATCC 17552) to nonpolarized and polarized thin films of gold. AB - The adhesion of Pseudomonas fluorescens (ATCC 17552) to nonpolarized and negatively polarized thin films of gold was studied in situ by contrast microscopy using a thin-film electrochemical flow cell. The influence of the electrochemical potential was evaluated at two different ionic strengths (0.01 and 0.1 M NaCl; pH 7) under controlled flow. Adhesion to nonpolarized gold surfaces readily increased with the time of exposition at both ionic-strength values. At negative potentials (-0.2 and -0.5 V [Ag/AgCl-KCl saturated [sat.]]), on the other hand, bacterial adhesion was strongly inhibited. At 0.01 M NaCl, the inhibition was almost total at both negative potentials, whereas at 0.1 M NaCl the inhibition was proportional to the magnitude of the potential, being almost total at -0.5 V. The existence of reversible adhesion was investigated by carrying out experiments under stagnant conditions. Reversible adhesion was observed only at potential values very close to the potential of zero charge of the gold surface (0.0 V [Ag/AgCl-KCl sat.]) at a high ionic strength (0.1 M NaCl). Theoretical calculations of the Derjaguin-Landau-Verwey-Overbeek (DLVO) interaction energy for the bacteria-gold interaction were in good agreement with experimental results at low ionic strength (0.01 M). At high ionic strength (0.1 M), deviations from DLVO behavior related to the participation of specific interactions were observed, when surfaces were polarized to negative potentials. PMID- 11425741 TI - Species-specific PCR for identification of common contaminant mollicutes in cell culture. AB - Mycoplasma arginini, M. fermentans, M. hyorhinis, M. orale, and Acholeplasma laidlawii are the members of the class Mollicutes most commonly found in contaminated cell cultures. Previous studies have shown that the published PCR primer pairs designed to detect mollicutes in cell cultures are not entirely specific. The 16S rRNA gene, the 16S-23S rRNA intergenic spacer region, and the 5' end of the 23S rRNA gene, as a whole, are promising targets for design of mollicute species-specific primer pairs. We analyzed the 16S rRNA genes, the 16S 23S rRNA intergenic spacer regions, and the 5' end of the 23S rRNA genes of these mollicutes and developed PCR methods for species identification based on these regions. Using high melting temperatures, we developed a rapid-cycle PCR for detection and identification of contaminant mollicutes. Previously published, putative mollicute-specific primers amplified DNA from 73 contaminated cell lines, but the presence of mollicutes was confirmed by species-specific PCR in only 60. Sequences of the remaining 13 amplicons were identified as those of gram positive bacterial species. Species-specific PCR primers are needed to confirm the presence of mollicutes in specimens and for identification, if required. PMID- 11425742 TI - Characterization of neutralizing antibodies and identification of neutralizing epitope mimics on the Clostridium botulinum neurotoxin type A. AB - Clostridium botulinum neurotoxin type A (BTx-A) is known to inhibit the release of acetylcholine at the neuromuscular junctions and synapses and to cause neuroparalysis and death. In this study, we have identified two monoclonal antibodies, BT57-1 and BT150-3, which protect ICR mice against lethal doses of BTx-A challenge. The neutralizing activities for BT57-1 and BT150-3 were 10(3) and 10(4) times the 50% lethal dose, respectively. Using immunoblotting analysis, BT57-1 was recognized as a light chain and BT150-3 was recognized as a heavy chain of BTx-A. Also, applying the phage display method, we investigated the antibodies' neutralizing B-cell epitopes. These immunopositive phage clones displayed consensus motifs, Asp-Pro-Leu for BT57-1 and Cys-X-Asp-Cys for BT150. The synthetic peptide P4M (KGTFDPLQEPRT) corresponded to the phage-displayed peptide selected by BT57-1 and was able to bind the antibodies specifically. This peptide was also shown by competitive inhibition assay to be able to inhibit phage clone binding to BT57-1. Aspartic acid (D(5)) in P4M was crucial to the binding of P4M to BT57-1, since its binding activity dramatically decreased when it was changed to lysine (K(5)). Finally, immunizing mice with the selected phage clones elicited a specific humoral response against BTx-A. These results suggest that phage-displayed random-peptide libraries are useful in identifying the neutralizing epitopes of monoclonal antibodies. In the future, the identification of the neutralizing epitopes of BTx-A may provide important information for the identification of the BTx-A receptor and the design of a BTx-A vaccine. PMID- 11425743 TI - Dual-bioaugmentation strategy to enhance remediation of cocontaminated soil. AB - Although metals are thought to inhibit the ability of microorganisms to degrade organic pollutants, several microbial mechanisms of resistance to metal are known to exist. This study examined the potential of cadmium-resistant microorganisms to reduce soluble cadmium levels to enhance degradation of 2,4 dichlorophenoxyacetic acid (2,4-D) under conditions of cocontamination. Four cadmium-resistant soil microorganisms were examined in this study. Resistant up to a cadmium concentration of 275 microg ml(-1), these isolates represented the common soil genera Arthrobacter, Bacillus, and Pseudomonas. Isolates Pseudomonas sp. strain H1 and Bacillus sp. strain H9 had a plasmid-dependent intracellular mechanism of cadmium detoxification, reducing soluble cadmium levels by 36%. Isolates Arthrobacter strain D9 and Pseudomonas strain I1a both produced an extracellular polymer layer that bound and reduced soluble cadmium levels by 22 and 11%, respectively. Although none of the cadmium-resistant isolates could degrade 2,4-D, results of dual-bioaugmentation studies conducted with both pure culture and laboratory soil microcosms showed that each of four cadmium-resistant isolates supported the degradation of 500-microg ml(-1) 2,4-D by the cadmium sensitive 2,4-D degrader Ralstonia eutropha JMP134. Degradation occurred in the presence of up to 24 microg of cadmium ml(-1) in pure culture and up to 60 microg of cadmium g(-1) in amended soil microcosms. In a pilot field study conducted with 5-gallon soil bioreactors, the dual-bioaugmentation strategy was again evaluated. Here, the cadmium-resistant isolate Pseudomonas strain H1 enhanced degradation of 2,4-D in reactors inoculated with R. eutropha JMP134 in the presence of 60 microg of cadmium g(-1). Overall, dual bioaugmentation appears to be a viable approach in the remediation of cocontaminated soils. PMID- 11425744 TI - Cross-resistance and stability of resistance to Bacillus thuringiensis toxin Cry1C in diamondback moth. AB - We tested toxins of Bacillus thuringiensis against larvae from susceptible, Cry1C resistant, and Cry1A-resistant strains of diamondback moth (Plutella xylostella). The Cry1C-resistant strain, which was derived from a field population that had evolved resistance to B. thuringiensis subsp. kurstaki and B. thuringiensis subsp. aizawai, was selected repeatedly with Cry1C in the laboratory. The Cry1C resistant strain had strong cross-resistance to Cry1Ab, Cry1Ac, and Cry1F, low to moderate cross-resistance to Cry1Aa and Cry9Ca, and no cross-resistance to Cry1Bb, Cry1Ja, and Cry2A. Resistance to Cry1C declined when selection was relaxed. Together with previously reported data, the new data on the cross resistance of a Cry1C-resistant strain reported here suggest that resistance to Cry1A and Cry1C toxins confers little or no cross-resistance to Cry1Bb, Cry2Aa, or Cry9Ca. Therefore, these toxins might be useful in rotations or combinations with Cry1A and Cry1C toxins. Cry9Ca was much more potent than Cry1Bb or Cry2Aa and thus might be especially useful against diamondback moth. PMID- 11425745 TI - The mannose-sensitive hemagglutinin of Vibrio cholerae promotes adherence to zooplankton. AB - The bacterium Vibrio cholerae, the etiological agent of cholera, is often found attached to plankton, a property that is thought to contribute to its environmental persistence in aquatic habitats. The V. cholerae O1 El Tor biotype and V. cholerae O139 strains produce a surface pilus termed the mannose-sensitive hemagglutinin (MSHA), whereas V. cholerae O1 classical biotype strains do not. Although V. cholerae O1 classical does not elaborate MSHA, the gene is present and expressed at a level comparable to that of the other strains. Since V. cholerae O1 El Tor and V. cholerae O139 have displaced V. cholerae O1 classical as the major epidemic strains over the last fifteen years, we investigated the potential role of MSHA in mediating adherence to plankton. We found that mutation of mshA in V. cholerae O1 El Tor significantly diminished, but did not eliminate, adherence to exoskeletons of the planktonic crustacean Daphnia pulex. The effect of the mutation was more pronounced for V. cholerae O139, essentially eliminating adherence. Adherence of the V. cholerae O1 classical mshA mutant was unaffected. The results suggest that MSHA is a factor contributing to the ability of V. cholerae to adhere to plankton. The results also showed that both biotypes of V. cholerae O1 utilize factors in addition to MSHA for zooplankton adherence. The expression of MSHA and these additional, yet to be defined, adherence factors differ in a serogroup- and biotype-specific manner. PMID- 11425746 TI - Microbial populations associated with treatment of an industrial dye effluent in an anaerobic baffled reactor. AB - Fluorescent in situ hybridization (FISH) using 16S and 23S rRNA-targeted probes together with construction of an archaeal 16S ribosomal DNA (rDNA) clone library was used to characterize the microbial populations of an anaerobic baffled reactor successfully treating industrial dye waste. Wastewater produced during the manufacture of food dyes containing several different azo and other dye compounds was decolorized and degraded under sulfidogenic and methanogenic conditions. Use of molecular methods to describe microbial populations showed that a diverse group of Bacteria and Archaea was involved in this treatment process. FISH enumeration showed that members of the gamma subclass of the class Proteobacteria and bacteria in the Cytophaga-Flexibacter-Bacteroides phylum, together with sulfate-reducing bacteria, were prominent members of a mixed bacterial population. A combination of FISH probing and analysis of 98 archaeal 16S rDNA clone inserts revealed that together with the bacterial population, a methanogenic population dominated by Methanosaeta species and containing species of Methanobacterium and Methanospirillum and a relatively unstudied methanogen, Methanomethylovorans hollandica, contributed to successful anaerobic treatment of the industrial waste. We suggest that sulfate reducers, or more accurately sulfidogenic bacteria, together with M. hollandica contribute considerably to the treatment process through metabolism of dye-associated sulfonate groups and subsequent conversion of sulfur compounds to carbon dioxide and methane. PMID- 11425747 TI - Identification of a small tetraheme cytochrome c and a flavocytochrome c as two of the principal soluble cytochromes c in Shewanella oneidensis strain MR1. AB - Two abundant, low-redox-potential cytochromes c were purified from the facultative anaerobe Shewanella oneidensis strain MR1 grown anaerobically with fumarate. The small cytochrome was completely sequenced, and the genes coding for both proteins were cloned and sequenced. The small cytochrome c contains 91 residues and four heme binding sites. It is most similar to the cytochromes c from Shewanella frigidimarina (formerly Shewanella putrefaciens) NCIMB400 and the unclassified bacterial strain H1R (64 and 55% identity, respectively). The amount of the small tetraheme cytochrome is regulated by anaerobiosis, but not by fumarate. The larger of the two low-potential cytochromes contains tetraheme and flavin domains and is regulated by anaerobiosis and by fumarate and thus most nearly corresponds to the flavocytochrome c-fumarate reductase previously characterized from S. frigidimarina to which it is 59% identical. However, the genetic context of the cytochrome genes is not the same for the two Shewanella species, and they are not located in multicistronic operons. The small cytochrome c and the cytochrome domain of the flavocytochrome c are also homologous, showing 34% identity. Structural comparison shows that the Shewanella tetraheme cytochromes are not related to the Desulfovibrio cytochromes c(3) but define a new folding motif for small multiheme cytochromes c. PMID- 11425748 TI - Impact of fumigants on soil microbial communities. AB - Agricultural soils are typically fumigated to provide effective control of nematodes, soilborne pathogens, and weeds in preparation for planting of high value cash crops. The ability of soil microbial communities to recover after treatment with fumigants was examined using culture-dependent (Biolog) and culture-independent (phospholipid fatty acid [PLFA] analysis and denaturing gradient gel electrophoresis [DGGE] of 16S ribosomal DNA [rDNA] fragments amplified directly from soil DNA) approaches. Changes in soil microbial community structure were examined in a microcosm experiment following the application of methyl bromide (MeBr), methyl isothiocyanate, 1,3-dichloropropene (1,3-D), and chloropicrin. Variations among Biolog fingerprints showed that the effect of MeBr on heterotrophic microbial activities was most severe in the first week and that thereafter the effects of MeBr and the other fumigants were expressed at much lower levels. The results of PLFA analysis demonstrated a community shift in all treatments to a community dominated by gram-positive bacterial biomass. Different 16S rDNA profiles from fumigated soils were quantified by analyzing the DGGE band patterns. The Shannon-Weaver index of diversity, H, was calculated for each fumigated soil sample. High diversity indices were maintained between the control soil and the fumigant-treated soils, except for MeBr (H decreased from 1.14 to 0.13). After 12 weeks of incubation, H increased to 0.73 in the MeBr-treated samples. Sequence analysis of clones generated from unique bands showed the presence of taxonomically unique clones that had emerged from the MeBr-treated samples and were dominated by clones closely related to Bacillus spp. and Heliothrix oregonensis. Variations in the data were much higher in the Biolog assay than in the PLFA and DGGE assays, suggesting a high sensitivity of PLFA analysis and DGGE in monitoring the effects of fumigants on soil community composition and structure. Our results indicate that MeBr has the greatest impact on soil microbial communities and that 1,3-D has the least impact. PMID- 11425749 TI - Microarray analysis of microbial virulence factors. AB - Hybridization with oligonucleotide microchips (microarrays) was used for discrimination among strains of Escherichia coli and other pathogenic enteric bacteria harboring various virulence factors. Oligonucleotide microchips are miniature arrays of gene-specific oligonucleotide probes immobilized on a glass surface. The combination of this technique with the amplification of genetic material by PCR is a powerful tool for the detection of and simultaneous discrimination among food-borne human pathogens. The presence of six genes (eaeA, slt-I, slt-II, fliC, rfbE, and ipaH) encoding bacterial antigenic determinants and virulence factors of bacterial strains was monitored by multiplex PCR followed by hybridization of the denatured PCR product to the gene-specific oligonucleotides on the microchip. The assay was able to detect these virulence factors in 15 Salmonella, Shigella, and E. coli strains. The results of the chip analysis were confirmed by hybridization of radiolabeled gene-specific probes to genomic DNA from bacterial colonies. In contrast, gel electrophoretic analysis of the multiplex PCR products used for the microarray analysis produced ambiguous results due to the presence of unexpected and uncharacterized bands. Our results suggest that microarray analysis of microbial virulence factors might be very useful for automated identification and characterization of bacterial pathogens. PMID- 11425750 TI - Nitrogen-fixing nodules with Ensifer adhaerens harboring Rhizobium tropici symbiotic plasmids. AB - Ensifer adhaerens is a soil bacterium that attaches to other bacteria and may cause lysis of these other bacteria. Based on the sequence of its small-subunit rRNA gene, E. adhaerens is related to Sinorhizobium spp. E. adhaerens ATCC 33499 did not nodulate Phaseolus vulgaris (bean) or Leucaena leucocephala, but with symbiotic plasmids from Rhizobium tropici CFN299 it formed nitrogen-fixing nodules on both hosts. The nodule isolates were identified as E. adhaerens isolates by growth on selective media. PMID- 11425751 TI - Toxic-metabolite-producing bacteria and fungus in an indoor environment. AB - Toxic-metabolite-emitting microbes were isolated from the indoor environment of a building where the occupant was suffering serious building-related ill-health symptoms. Toxic substances soluble in methanol and inhibitory to spermatozoa at <10 microg (dry weight) ml(-1) were found from six bacterial isolates and one fungus. The substances from isolates of Bacillus simplex and from isolates belonging to the actinobacterial genera Streptomyces and Nocardiopsis were mitochondriotoxic. These substances dissipated the mitochondrial membrane potential (Deltapsi) of boar spermatozoa. The substances from the Streptomyces isolates also swelled the mitochondria. The substances from isolates of Trichoderma harzianum Rifai and Bacillus pumilus damaged the cell membrane barrier function of sperm cells. PMID- 11425752 TI - Reductive precipitation of gold by dissimilatory Fe(III)-reducing bacteria and archaea. AB - Studies with a diversity of hyperthermophilic and mesophilic dissimilatory Fe(III)-reducing Bacteria and Archaea demonstrated that some of these organisms are capable of precipitating gold by reducing Au(III) to Au(0) with hydrogen as the electron donor. These studies suggest that models for the formation of gold deposits in both hydrothermal and cooler environments should consider the possibility that dissimilatory metal-reducing microorganisms can reductively precipitate gold from solution. PMID- 11425753 TI - Cyt1Ab1 and Cyt2Ba1 from Bacillus thuringiensis subsp. medellin and B. thuringiensis subsp. israelensis Synergize Bacillus sphaericus against Aedes aegypti and resistant Culex quinquefasciatus (Diptera: Culicidae). AB - The interaction of two cytolytic toxins, Cyt1Ab from Bacillus thuringiensis subsp. medellin and Cyt2Ba from Bacillus thuringiensis subsp. israelensis, with Bacillus sphaericus was evaluated against susceptible and resistant Culex quinquefasciatus and the nonsensitive species Aedes aegypti. Mixtures of B. sphaericus with either cytolytic toxin were synergistic, and B. sphaericus resistance in C. quinquefasciatus was suppressed from >17,000- to 2-fold with a 3:1 mixture of B. sphaericus and Cyt1Ab. This trait may prove useful for combating insecticide resistance and for improving the activity of microbial insecticides. PMID- 11425754 TI - Distribution, isolation, host specificity, and diversity of cyanophages infecting marine Synechococcus spp. in river estuaries. AB - The abundance of cyanophages infecting marine Synechococcus spp. increased with increasing salinity in three Georgia coastal rivers. About 80% of the cyanophage isolates were cyanomyoviruses. High cross-infectivity was found among the cyanophages infecting phycoerythrin-containing Synechococcus strains. Cyanophages in the river estuaries were diverse in terms of their morphotypes and genotypes. PMID- 11425755 TI - Development of a quantitative competitive PCR assay for detection and quantification of Escherichia coli O157:H7 cells. AB - A quantitative competitive PCR (QC-PCR) assay was developed to detect and quantify Escherichia coli O157:H7 cells. From 10(3) to 10(8) CFU of E. coli O157:H7 cells/ml was quantified in broth or skim milk, and cell densities predicted by QC-PCR were highly related to viable cell counts (r(2) = 0.99 and 0.93, respectively). QC-PCR has potential for quantitative detection of pathogenic bacteria in foods. PMID- 11425756 TI - Rapid approach to determine rrn arrangement in Salmonella serovars. AB - A PCR method was developed by which to rapidly and accurately determine the rrn arrangement of Salmonella enterica serovars. Primers were designed to the genomic regions flanking each of the seven rrn operons. PCR analysis using combinations of these primers will distinguish each of the possible arrangements of the rrn skeleton. PMID- 11425757 TI - True chemotaxis in oxygen gradients of the sulfur-oxidizing bacterium Thiovulum majus. AB - Observations of free-swimming Thiovulum majus cells show that these bacteria exhibit a phobic response as well as true chemotaxis in oxygen gradients. Both phenomena of their chemotactic behavior are integrated into a single model of helical klinotaxis, which is demonstrated by computer simulations. PMID- 11425758 TI - Organization and regulation of meta cleavage pathway genes for toluene and o xylene derivative degradation in Pseudomonas stutzeri OX1. AB - Pseudomonas stutzeri OX1 meta pathway genes for toluene and o-xylene catabolism were analyzed, and loci encoding phenol hydroxylase, catechol 2,3-dioxygenase, 2 hydroxymuconate semialdehyde dehydrogenase, and 2-hydroxymuconate semialdehyde hydrolase were mapped. Phenol hydroxylase converted a broad range of substrates, as it was also able to transform the nongrowth substrates 2,4-dimethylphenol and 2,5-dimethylphenol into 3,5-dimethylcatechol and 3,6-dimethylcatechol, respectively, which, however, were not cleaved by catechol 2,3-dioxygenase. The identified gene cluster displayed a gene order similar to that of the Pseudomonas sp. strain CF600 dmp operon for phenol catabolism and was found to be coregulated by the tou operon activator TouR. A hypothesis about the evolution of the toluene and o-xylene catabolic pathway in P. stutzeri OX1 is discussed. PMID- 11425759 TI - Decrease in Cryptosporidium parvum oocyst infectivity in vitro by using the membrane filter dissolution method for recovering oocysts from water samples. AB - Exposure of Cryptosporidium parvum oocysts to solutions used for cellulose acetate membrane (CAM) dissolution filtration reduced their infectivity in HCT-8 cells. Ethanol (95% [vol/vol] and 70% [vol/vol]) alone and short exposure times to acetone decreased infectivity. These findings contrast with similar experiments using excystation assays and infectivity in mice. PMID- 11425760 TI - Congruent phylogenies of most common small-subunit rRNA and dissimilatory sulfite reductase gene sequences retrieved from estuarine sediments. AB - The diversity of sulfate-reducing bacteria (SRB) in brackish sediment was investigated using small-subunit rRNA and dissimilatory sulfite reductase (DSR) gene clone libraries and cultivation. The phylogenetic affiliation of the most commonly retrieved clones for both genes was strikingly similar and produced Desulfosarcina variabilis-like sequences from the inoculum but Desulfomicrobium baculatum-like sequences from a high dilution in natural media. Related organisms were subsequently cultivated from the site. PCR bias appear to be limited (or very similar) for the two primersets and target genes. However, the DSR primers showed a much higher phylogenetic specificity. DSR gene analysis is thus a promising and specific approach for investigating SRB diversity in complex habitats. PMID- 11425761 TI - Exo-arabinanase of Penicillium chrysogenum able to release arabinobiose from alpha-1,5-L-arabinan. AB - An exo-arabinanase, designated Abnx, was purified from a culture filtrate of Penicillium chrysogenum 31B by ammonium sulfate precipitation, anion-exchange chromatography, and hydrophobic chromatography. Abnx had an apparent molecular mass of 47 kDa. The enzyme released only arabinobiose from the nonreducing terminus of alpha-1,5-L-arabinan and showed no activity towards p-nitrophenyl alpha-L-arabinofuranoside and alpha-1,5-L-arabinofuranobiose. Abnx is the first enzyme with this mode of action. PMID- 11425762 TI - Effect of high-rate algal ponds on viability of Cryptosporidium parvum oocysts. AB - The physicochemical conditions of high-rate algal ponds were responsible for a more than 97% reduction in the infectivity of Cryptosporidium parvum oocysts in neonatal mice. The use of semipermeable bags of cellulose showed that pH, ammonia, and/or light seems to be a major factor for the inactivation of oocysts in wastewater, supporting the importance of alga-based systems for safer reuse of treated wastewater. PMID- 11425763 TI - Incidence of Klebsiella species in surface waters and their expression of virulence factors. AB - To investigate the occurrence of different Klebsiella spp. in aquatic environments, a total of 208 samples of natural surface waters was examined. From half (53%) of these samples, 123 Klebsiella strains were isolated, the most common species being Klebsiella pneumoniae. A comparison of these isolates to a group of 207 clinical K. pneumoniae isolates demonstrated that water isolates of K. pneumoniae, unlike those of K. oxytoca and K. planticola, are as capable as clinical isolates of expressing putative virulence factors such as serum resistance and capsular polysaccharides, pili, and siderophores. PMID- 11425764 TI - Microbiological analysis of tube-well water in a rural area of Bangladesh. AB - Five tube-wells in Matlab, Bangladesh, were selected for analysis of selected biophysicochemical parameters. The results showed that all tube-well water samples contained zooplankton and bacteria. Results for some of the parameters were outside the accepted limits recommended by the World Health Organization for drinking water. It is concluded that water from tube-wells should be treated if used as drinking water. PMID- 11425765 TI - Deficiency of tissue factor pathway inhibitor promotes atherosclerosis and thrombosis in mice. AB - BACKGROUND: Tissue factor initiates blood coagulation after atherosclerotic plaque disruption. Tissue factor pathway inhibitor (TFPI) inhibits tissue factor activity and may reduce thrombus formation in this setting. We evaluated the effect of heterozygous TFPI deficiency on the development of atherosclerosis and thrombosis in atherosclerosis-prone mice. METHODS AND RESULTS: Mice with a combined heterozygous TFPI deficiency and homozygous apolipoprotein E deficiency (TFPI(+/-)/apoE(-/-)) were generated by crossbreeding, and they were analyzed for atherosclerosis throughout the vascular tree. Compared with mice with a normal TFPI genotype (TFPI(+/+)/apoE(-/-)), mice with a TFPI deficiency exhibited a greater atherosclerotic burden involving the carotid and common iliac arteries. Staining for active tissue factor within the plaque revealed more activity in TFPI(+/-)/apoE(-/-) mice compared with TFPI(+/+)/apoE(-/-) mice. Consistent with increased plaque tissue factor activity, the time to occlusive thrombosis after photochemical carotid plaque injury was significantly decreased in TFPI(+/ )/apoE(-/-) mice. CONCLUSIONS: These observations indicate that TFPI protects from atherosclerosis and is an important regulator of the thrombosis that occurs in the setting of atherosclerosis. PMID- 11425766 TI - High-dose recombinant apolipoprotein A-I(milano) mobilizes tissue cholesterol and rapidly reduces plaque lipid and macrophage content in apolipoprotein e-deficient mice. Potential implications for acute plaque stabilization. AB - BACKGROUND: Repeated doses of recombinant apolipoprotein A-I(Milano) phospholipid complex (apoA-I(m)) reduce atherosclerosis and favorably change plaque composition in rabbits and mice. In this study, we tested whether a single high dose of recombinant apoA-I(m) could rapidly mobilize tissue cholesterol and reduce plaque lipid and macrophage content in apoE-deficient mice. METHODS AND RESULTS: High cholesterol-fed, 26-week-old apoE-deficient mice received a single intravenous injection of saline (n=16), 1080 mg/kg dipalmitoylphosphatidylcholine (DPPC; n=14), or 400 mg/kg of recombinant apoA-I(m) complexed with DPPC (1:2.7 weight ratio; n=18). Blood was sampled before and 1 and 48 hours after injection, and aortic root plaques were evaluated for lipid content and macrophage content after oil-red O and immunostaining, respectively. One hour after injection, the plasma cholesterol efflux-promoting capacity was nearly 2-fold higher in recombinant apoA-I(m)-treated mice compared with saline and DPPC-treated mice (P<0.01). Compared with baseline values, serum free cholesterol, an index of tissue cholesterol mobilization, increased 1.6-fold by 1 hour after recombinant apoA-I(m) injection, and it remained significantly elevated at 48 hours (P<0.01). Mice receiving recombinant apoA-I(m) had 40% to 50% lower lipid content (P<0.01) and 29% to 36% lower macrophage content (P<0.05) in their plaques compared with the saline- and DPPC-treated mice, respectively. CONCLUSIONS: A single high dose of recombinant apoA-I(m) rapidly mobilizes tissue cholesterol and reduces plaque lipid and macrophage content in apoE-deficient mice. These findings suggest that this strategy could rapidly change plaque composition toward a more stable phenotype. PMID- 11425767 TI - Genetic and environmental contributions to platelet aggregation: the Framingham heart study. AB - BACKGROUND: Platelet aggregation plays an important role in arterial thrombosis in coronary heart disease, stroke, and peripheral arterial disease. However, the contribution of genetic versus environmental influences on interindividual variation in platelet aggregability is poorly characterized. METHODS AND RESULTS: We studied the heritability of platelet aggregation responses in 2413 participants in the Framingham Heart Study. The threshold concentrations of epinephrine and ADP required to produce biphasic platelet aggregation and collagen lag time were determined. Mixed-model linear regression was used to calculate correlation coefficients within sibships and within spouse pairs. Variance and covariance component methods were used to estimate the proportion of platelet aggregation attributable to measured covariates versus additive genetic effects. After accounting for environmental covariates, the adjusted sibling correlations for epinephrine, ADP, and collagen lag time were 0.24, 0.22, and 0.31, respectively (P=0.0001 for each). In contrast, adjusted correlations for spouse-pairs were -0.01, 0.05, and -0.02, respectively (all P>0.30). The estimated heritabilities were 0.48, 0.44, and 0.62, respectively. Measured covariates accounted for only 4% to 7% of the overall variance in platelet aggregation, and heritable factors accounted for 20% to 30%. The platelet glycoprotein IIIa Pl(A2) polymorphism and the fibrinogen Hind III beta-148 polymorphism contributed <1% to the overall variance. CONCLUSIONS: In our large, population-based sample, heritable factors play a major role in determining platelet aggregation, and measured covariates play a lesser role. Future studies are warranted to identify the key genetic variants that regulate platelet function and to lay the groundwork for rational pharmacogenetic approaches. PMID- 11425768 TI - Excessive urinary albumin levels are associated with future cardiovascular mortality in postmenopausal women. AB - BACKGROUND: Microalbuminuria is an early predictor of cardiovascular morbidity and mortality, in both diabetic patients and hypertensive patients. Little is known about the relation of microalbuminuria to cardiovascular disease in women of the general population. METHODS AND RESULTS: We have studied the relation of urinary albumin levels to cardiovascular mortality in a cohort study of 12 239 postmenopausal women living in Utrecht, the Netherlands. The initial age was between 52 and 67 years. Women were followed on vital status between 1976 and 1995 (168 513 women-years). Albumin was determined in the urine of 561 cases and 557 controls. Data were analyzed by using a nested case-control design. The cardiovascular mortality rate (95% CI) for women who were in the highest quintile of urinary albumin levels was 13.2/1000 years (8.1 to 20.9) compared with 2.6/1000 years (2.3 to 3.1) in women without detectable urinary albumin. The age adjusted rate ratio (95% CI) between these groups was 4.4 (2.6 to 7.6). CONCLUSIONS: This is the first large cohort study that confirms a predictive role of urinary albumin for the risk of future cardiovascular mortality independent of hypertension and diabetes. Our findings support the hypothesis that microalbuminuria is a reflection of vascular damage and a marker of early arterial disease in women from the general population. PMID- 11425769 TI - Biological profiles in subjects with recurrent acute coronary events compared with subjects with long-standing stable angina. AB - BACKGROUND: At one end of the clinical spectrum of coronary artery disease (CAD) are subjects who have had repeated acute ischemic events, and at the other end are those with long-standing angina who have never been unstable. This study tests the hypothesis that a specific biological profile can distinguish these 2 extreme groups and predict acute coronary events. METHODS AND RESULTS: Blood levels of lipoprotein(a), homocysteine, tissue plasminogen activator, plasminogen activator inhibitor-1, C-reactive protein (CRP), fibrinogen, and von Willebrand factor were compared in 3 groups of 50 subjects each: (1) those with previous multiple acute coronary events, (2) age-matched subjects with >/=3 years of stable angina and no prior acute coronary events, and (3) matched controls without evidence of atherosclerotic disease and a normal coronary angiogram. All subjects were followed for 4.0 years. Lipoprotein(a), homocysteine, tissue plasminogen activator, and plasminogen activator inhibitor-1 were similar in both CAD groups and significantly higher than in the control group. However, compared with subjects with long-standing stable angina, those with previous multiple coronary events had higher values of CRP (5.7+/-5.4 versus 3.0+/-5.2 mg/L, P=0.012), fibrinogen (3.38+/-0.75 versus 2.92+/-0.64 g/L, P=0.001), and von Willebrand factor (1.60+/-0.55 versus 1.25+/-0.36 U/mL, P=0.0003). On follow-up, myocardial infarction and unstable angina occurred in 42% of the group with multiple events, 4% of the stable angina group (P<0.0001), and none of the control subjects. In the 100 patients with CAD, CRP was 4.9 mg/L in those with and 1.8 mg/L in those without new instability (P<0.0001). In a multivariate analysis, only CRP distinguished those with follow-up acute coronary events (adjusted odds ratio 5.9, 95% CI 2.0 to 17.9; P=0.002). A baseline CRP >3.5 mg/L had a relative risk of 7.6 (2.6 to 21.7, P=0.0002) for subsequent acute events. CONCLUSIONS: An inflammatory biological profile distinguished patients with previous multiple acute coronary events from those with long-standing stable angina and predicted acute coronary instability. PMID- 11425770 TI - Aspirin, warfarin, or the combination for secondary prevention of coronary events in patients with acute coronary syndromes and prior coronary artery bypass surgery. AB - BACKGROUND: Patients with a non-ST-elevation acute coronary syndrome and prior CABG are at high risk of a recurrent ischemic event despite aspirin therapy. This trial investigated the potential benefit of secondary prevention with warfarin. METHODS AND RESULTS: In a double-blind randomized trial, 135 patients with unstable angina or non-ST-segment elevation myocardial infarction, with prior CABG, and who were poor candidates for a revascularization procedure received therapy with aspirin and placebo+warfarin, warfarin and placebo+aspirin, or aspirin and warfarin for 12 months. Warfarin was titrated to an international normalized ratio of 2.0 to 2.5. The primary end point (death or myocardial infarction or unstable angina requiring hospitalization 1 year after randomization) occurred in 14.6% of the patients in the warfarin-alone group, in 11.5% of patients in the aspirin-alone group, and in 11.3% of patients randomized to the combination therapy (P=0.76). Subgroup analyses by risk features provided no indications that warfarin alone or in combination with aspirin could be of benefit over aspirin alone. Bleeding was more frequent in the 2 groups of patients administered warfarin. CONCLUSIONS: Moderate-intensity oral anticoagulation alone or combined with low-dose aspirin does not appear to be superior to low-dose aspirin in the prevention of recurrent ischemic events in patients with non-ST-elevation acute coronary syndromes and previous CABG. PMID- 11425771 TI - Dispersion of ventricular depolarization-repolarization: a noninvasive marker for risk stratification in arrhythmogenic right ventricular cardiomyopathy. AB - BACKGROUND: We retrospectively investigated the value of clinical and ECG findings as well as QT-QRS dispersion in predicting the risk of sudden death in patients with arrhythmogenic right ventricular cardiomyopathy (ARVC). METHODS AND RESULTS: Duration and interlead variability of the QT interval and QRS complex were measured manually from standard ECGs in 20 sudden death victims with ARVC diagnosed at autopsy (group I), in 20 living ARVC patients with sustained ventricular tachycardia (group II), in 20 living ARVC patients with /=40 ms had a sensitivity and specificity of 90% and 77%, respectively; QT dispersion >65 ms, 85% and 75%, respectively; negative T wave beyond V(1), 85% and 42%, respectively; and syncope, 40% and 90%, respectively. CONCLUSIONS: QRS dispersion (>/=40 ms) was the strongest independent predictor of sudden death in ARVC. Syncope, QT dispersion >65 ms, and negative T wave beyond V(1) refined arrhythmic risk stratification in these patients. PMID- 11425772 TI - Relationship between pacemaker dependency and the effect of pacing mode on cardiovascular outcomes. AB - BACKGROUND: A recently completed trial, the Canadian Trial of Physiological Pacing (CTOPP), showed that physiological pacing did not significantly reduce mortality, stroke, or heart failure hospitalization, but it did show that atrial fibrillation occurred less frequently in patients with physiological pacing. Many pacemaker patients experience only transient bradyarrhythmias with an adequate unpaced heart rate (UHR) and are not pacemaker-dependent. The purpose of the present analysis was to determine if pacemaker-dependent patients have an increased benefit from physiological pacing compared with non-pacemaker-dependent patients. METHODS AND RESULTS: Of 2568 patients included in the CTOPP trial, 2244 patients had a pacemaker dependency test performed at the first follow-up visit. The yearly event rate of cardiovascular death or stroke steadily increased with decreasing UHR in the ventricular pacing group, but it remained constant in the physiological pacing group. When the patients were subdivided to UHR 60 bpm, there was an interaction between pacing mode treatment and UHR subgroup. The Kaplan-Meier plot confirmed a physiological pacing advantage only in the UHR /=2 fold (ANOVA, P=0.002) and >/=3-fold (P<0.0001) higher after ST than after BA, respectively. Active MMP9 was detected only after ST. Although the increases in MMP2 mRNA levels were of similar magnitudes after ST and BA, pro-MMP2 activity was slightly higher 7 and 30 days after ST, and MMP2 activity was >/=2-fold higher 7 to 60 days after ST (P=0.002). No difference in TIMP expression was observed between stented and balloon-injured arteries. Cellular distributions of MMPs and TIMP1 were similar after ST and BA. Early inflammatory cell recruitment and 30-day intimal growth were more severe after ST. CONCLUSIONS: Stent implantation results in more intense and sustained expression of MMP9 and activation of MMP2 than balloon angioplasty. PMID- 11425779 TI - Angiotensin-converting enzyme inhibition attenuates hypofibrinolysis and reduces cardiac perivascular fibrosis in genetically obese diabetic mice. AB - BACKGROUND: Obesity and insulin resistance are associated with accelerated macrovascular and microvascular coronary disease, cardiomyopathic phenomena, and increased concentrations and activity in blood of plasminogen activator inhibitor type 1 (PAI-1), the primary physiological inhibitor of fibrinolysis. METHODS AND RESULTS: To determine whether hypofibrinolysis in blood and tissues and its potential sequelae could be attenuated pharmacologically, we studied genetically modified obese mice. By 10 weeks of age, obese mice exhibited increases in left ventricular weight and glucose and immunoreactive insulin in blood. PAI-1 activity in blood measured spectrophotometrically was significantly elevated as well. The difference compared with values in lean controls widened by 20 weeks of age. Perivascular fibrosis in coronary arterioles and small coronary arteries was evident in obese mice 10 and 20 weeks of age, paralleling increases in PAI-1 and tissue factor expression evident by immunohistochemical image analysis, in situ hybridization, and reverse transcription-polymerase chain reaction. Inhibition of ACE activity initiated in obese mice 10 weeks of age and continued for 20 weeks arrested the increase in PAI-1 activity in blood and in cardiac PAI-1 and tissue factor mRNA as well as coronary perivascular fibrosis. CONCLUSIONS: Thus, inhibition of proteo(fibrino)lysis and augmented tissue factor expression in the heart precede and may contribute to the coronary perivascular fibrosis seen with obesity and insulin resistance. Furthermore, inhibition of ACE activity can attenuate all 3 phenomena. PMID- 11425780 TI - Endothelin 1 type a receptor antagonism prevents vascular dysfunction and hypertension induced by 11beta-hydroxysteroid dehydrogenase inhibition: role of nitric oxide. AB - BACKGROUND: The enzyme 11beta-hydroxysteroid dehydrogenase (11beta-HSD) prevents inappropriate activation of the nonselective mineralocorticoid receptors by glucocorticoids. Renal activity of 11beta-HSD is decreased in patients with apparent mineralocorticoid excess (SAME), licorice-induced hypertension, and essential hypertension. Although expressed in vascular cells, the role of 11beta HSD in the regulation of vascular tone remains to be determined. METHODS AND RESULTS: lycyrrhizic acid (GA; 50 mg/kg IP, twice daily for 7 days) caused a significant inhibition of 11beta-HSD activity and induced hypertension in Wistar Kyoto rats (157 versus 127 mm Hg in controls; P<0.01). After 11beta-HSD inhibition, aortic endothelial nitric oxide (NO) synthase (eNOS) protein content, nitrate tissue levels, and acetylcholine-induced release of NO were blunted (all P<0.05 versus controls). In contrast, vascular prepro-endothelin (ET)-1 gene expression, ET-1 protein levels, and vascular reactivity to ET-1 were enhanced by GA treatment (P<0.05 versus controls). Chronic ET(A) receptor blockade with LU135252 (50 mg. kg(-1). d(-1)) normalized blood pressure, ET-1 tissue content, vascular reactivity to ET-1, vascular eNOS protein content, and nitrate tissue levels and improved NO-mediated endothelial function in GA-treated rats (P<0.05 to 0.01 versus untreated and verapamil-treated controls). In human endothelial cells, GA increased production of ET-1 in the presence of corticosterone, which indicates that activation of the vascular ET-1 system by 11beta-HSD inhibition can occur independently of changes in blood pressure but is dependent on the presence of glucocorticoids. CONCLUSIONS: Chronic ET(A) receptor blockade normalizes blood pressure, prevents upregulation of vascular ET-1, and improves endothelial dysfunction in 11beta-HSD inhibitor-induced hypertension and may emerge as a novel therapeutic approach in cardiovascular disease associated with reduced 11beta-HSD activity. PMID- 11425782 TI - Evaluation of the culprit plaque and the physiological significance of coronary atherosclerotic narrowings. PMID- 11425781 TI - Long-term effect of N-acetyl-seryl-aspartyl-lysyl-proline on left ventricular collagen deposition in rats with 2-kidney, 1-clip hypertension. AB - BACKGROUND: N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) is a natural inhibitor of pluripotent hematopoietic stem cell proliferation. Ac-SDKP plasma concentration is increased 5-fold after angiotensin-converting enzyme inhibition. Here we studied the effect of Ac-SDKP on monocyte/macrophage infiltration, fibroblast proliferation, and collagen deposition in the rat heart in renovascular hypertension. METHODS AND RESULTS: We investigated whether long-term Ac-SDKP administration would prevent left ventricular (LV) hypertrophy and interstitial collagen deposition in rats with 2-kidney, 1-clip (2K-1C) hypertension. Ac-SDKP (400 microgram. kg(-1). d(-1)) did not affect development of hypertension. Mean blood pressure was similar in rats with 2K-1C hypertension whether they were given vehicle or Ac-SDKP and was higher than in controls. Both LV weight and cardiomyocyte size were significantly increased in rats with 2K-1C hypertension compared with controls and were unaffected by Ac-SDKP. Proliferating cell nuclear antigen- and monocyte/macrophage-positive cells were increased in the LV of 2K-1C hypertensive rats; this increase was significantly blunted by Ac SDKP (P<0.001). LV interstitial collagen fraction was also increased in 2K-1C hypertensive rats given vehicle (10.1+/-0.8%) compared with sham (5.3+/-0.1%, P<0.0001), and this increase was prevented by Ac-SDKP (5.4+/-0.4%, P<0.001). CONCLUSIONS: Ac-SDKP inhibited monocyte/macrophage infiltration, cell proliferation, and collagen deposition in the LV of hypertensive rats without affecting blood pressure or cardiac hypertrophy, suggesting that it may be partly responsible for the cardioprotective effect of angiotensin-converting enzyme inhibitors. PMID- 11425783 TI - Random research. AB - Advances in computing have combined with the rapid dissemination of treatment discoveries for diseases of public health importance to create pressure for accelerated promulgation of promising research results to the medical community. The 2 recent examples of the US Carvedilol Heart Failure program and the Evaluation of Losartan In the Elderly (ELITE) study demonstrate the importance of the prospective nature of research design, as well as the consequences of its abandonment. This article explains in nonmathematical terms the rationale for the tenet "first say what you will do, then do what you said" in sample-based research. PMID- 11425784 TI - Congenital complete absence of the pericardium. PMID- 11425785 TI - Cardiology patient page: warning signs of a heart attack. PMID- 11425787 TI - Cigarette smoking: how much worse can it get? PMID- 11425786 TI - Familial scimitar syndrome: three-dimensional visualization of anomalous pulmonary vein in young sisters. PMID- 11425788 TI - Augmentation of coronary blood flow with intra-aortic balloon pump counter pulsation. PMID- 11425789 TI - Predictors of atrial fibrillation after conventional and beating heart coronary surgery. PMID- 11425790 TI - Price of modernization of China. PMID- 11425792 TI - Giorgio Olivetti. PMID- 11425794 TI - The ins(ide) and out(side) of dolichyl phosphate biosynthesis and recycling in the endoplasmic reticulum. AB - The precursor oligosaccharide donor for protein N-glycosylation in eukaryotes, Glc3Man9GlcNAc(2)-P-P-dolichol, is synthesized in two stages on both leaflets of the rough endoplasmic reticulum (ER). There is good evidence that the level of dolichyl monophosphate (Dol-P) is one rate-controlling factor in the first stage of the assembly process. In the current topological model it is proposed that ER proteins (flippases) then mediate the transbilayer movement of Man-P-Dol, Glc-P Dol, and Man5GlcNAc(2)-P-P-Dol from the cytoplasmic leaflet to the lumenal leaflet. The rate of flipping of the three intermediates could plausibly influence the conversion of Man5GlcNAc(2)-P-P-Dol to Glc3Man(9)GlcNAc(2)-P-P-Dol in the second stage on the lumenal side of the rough ER. This article reviews the current understanding of the enzymes involved in the de novo biosynthesis of Dol P and other polyisoprenoid glycosyl carrier lipids and speculates about the role of membrane proteins and enzymes that could be involved in the transbilayer movement of the lipid intermediates and the recycling of Dol-P and Dol-P-P discharged during glycosylphosphatidylinositol anchor biosynthesis, N glycosylation, and O- and C-mannosylation reactions on the lumenal surface of the rough ER. PMID- 11425795 TI - Lectin-like proteins in model organisms: implications for evolution of carbohydrate-binding activity. AB - Classes of intracellular lectins that recognize core-type structures and mediate intracellular glycoprotein trafficking are present in vertebrates, model invertebrates such as Caenorhabditis elegans and Drosophila melanogaster, plants, and yeasts. Lectins that recognize more complex structures at the cell surface, such as C-type lectins and galectins, are also found in invertebrate organisms as well as vertebrates, but the functions of these proteins have evolved differently in different animal lineages. PMID- 11425796 TI - Human lung adenocarcinoma alpha1,3/4-L-fucosyltransferase displays two molecular forms, high substrate affinity for clustered sialyl LacNAc type 1 units as well as mucin core 2 sialyl LacNAc type 2 unit and novel alpha1,2-L-fucosylating activity. AB - Human lung tumor alpha1,3/4-L-fucosyltransferase (FT) was purified (2000-fold, 29% recovery) from 290 g of tissue by including a chromatography step on Affinity Gel-GDP. Two molecular forms (FTA, larger size carrying 15% alpha1,4-FT activity; FTB, the major form with 85% activity) were separated by further fractionation on a Sephacryl S-100 HR column. A difference in the electrophoretic mobilities of these two activities was also found on native polyacrylamide gel electrophoresis (PAGE). Both forms were devoid of typical alpha1,2-fucosylating activity but were associated with the novel alpha1,2-fucosylating ability of converting the Lewis a determinant to Lewis b. Based on percentage activity toward 2-O MeGalbeta1,3GlcNAcbeta-O-Bn, both forms exhibited the same extent of activity toward various acceptors, which included sulfated, sialylated, or methylated LacNAc type 1 or type 2 as well as mucin core 2 acceptors. However, FTA and FTB exhibited a difference in their ability to act on mucin core 2 3'-sialyl LacNAc (activities 24.2% and 40.8%, respectively, as compared to 2-O MeGalbeta1,3GlcNAcbeta-O-Bn). The unsubstituted LacNAc type 1 acceptors were 15 20 times as active as the corresponding LacNAc type 2 acceptors. The 3-O substitution on the beta1,4-linked Gal (methyl, sulfate, or sialyl) in mucin core 2 acceptors increased the efficiency of these acceptors five- to eightfold. The most efficient acceptor for FTA and FTB was 3-O-sulfoGalbeta1,3GlcNAcbeta-O-Al (K(m) 100 and 47 microM, respectively). The K(m) (mM) values for 2-O-methyl Galbeta1,3GlcNAcbeta-O-Bn and 3-O-sialyl Galbeta1,3GlcNAcbeta-O-Bn were 0.40 and 2.5 (FTA) and 0.16 and 0.67 (FTB), respectively. The 35-kDa glycoprotein ancrod (from Malayan pit viper venom) containing 36% complex N-glycans with the antennae NeuAcalpha2,3Galbeta1,3GlcNAcbeta- acted as the best macromolecular acceptor substrate (K(m): 45 microM), as examined with FTB. On desialylation the acceptor efficiency dropped to approximately 50% (K(m) for asialo ancrod: 167 microM). Sialylglycoproteins, such as carcinoembryonic antigen, fetuin, and bovine alpha(1)-acid glycoprotein, were better acceptors than asialo fetuin. On the contrary, fetuin triantennary glycopeptide containing predominantly NeuAcalpha2,3Galbeta1,4GlcNAcbeta- was only 55% active as compared to the asialo glycopeptide (K(m): 1.43 and 0.63 mM, respectively). Thus, the human lung tumor alpha1,3/4-L-FT has the potential to generate clustered sialyl Lewis a and Lewis b determinants in N-glycans and sialyl Lewis x determinant in mucin core 2 structures. PMID- 11425797 TI - Fucosylated human milk oligosaccharides vary between individuals and over the course of lactation. AB - Specific human milk oligosaccharides, especially fucosylated neutral oligosaccharides, protect infants against specific microbial pathogens. To study the concentrations of individual neutral oligosaccharides during lactation, a total of 84 milk samples were obtained from 12 women at 7 time periods during weeks 1-49 postpartum. The neutral oligosaccharides from each sample were isolated, perbenzoylated, resolved, and quantified by reversed-phase high performance liquid chromatography. The resultant oligosaccharide peaks, identified by co-elution with authentic standards and mass spectrometry, ranged in size from tri- to octasaccharides. The total concentration of oligosaccharides declined over the course of lactation; the mean concentration at 1 year was less than half that in the first few weeks postpartum. One of the 12 donors produced milk fucosyloligosaccharides that were essentially devoid of alpha1,2 linkages (but contained alpha1,3- and alpha1,4-linked fucose) until late in lactation, consistent with the nonsecretor phenotype. In milk samples from the remaining 11 donors, fucosyloligosaccharides containing alpha1,2-linked fucose were prevalent, and their profiles were distinct from those of fucosyloligosaccharides devoid of alpha1,2-linked fucose. The ratio of alpha1,2-linked oligosaccharide concentrations to oligosaccharides devoid of alpha1,2-linked fucose changed during the first year of lactation from 5:1 to 1:1. Furthermore, the absolute and the relative concentrations of individual oligosaccharides varied substantially, both between individual donors and over the course of lactation for each individual. The patterns of milk oligosaccharides among individuals suggest the existence of many genotype subpopulations. This variation in individual oligosaccharide concentrations suggests that the protective activities of human milk could also vary among individuals and during lactation. PMID- 11425798 TI - Characterization of N-glycans from mouse brain neural cell adhesion molecule. AB - The N-glycosylation pattern of the neural cell adhesion molecule (NCAM), isolated from brains of newborn mice, has been analyzed. Following digestion with trypsin, generated glycopeptides were fractionated by serial immunoaffinity chromatography using immobilized monoclonal antibodies specifically recognizing polysialic acid (PSA) units or the HNK1-carbohydrate epitope. Subsequent analyses of the resulting (glyco)peptides by Edman degradation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) revealed polysialylated glycans to be exclusively linked to glycosylation sites 5 (Asn(431)) and 6 (Asn(460)), whereas glycans carrying the HNK1-epitope could be assigned to sites 2 (Asn(297)), 5, 6, and, to a lesser extent, site 3 (Asn(329)). PSA-, HNK1-, and non-PSA/HNK1-glycan fractions were characterized by carbohydrate constituent and methylation analyses as well as MALDI-TOF-MS in conjunction with chromatographic fractionation techniques. The results revealed that the core structures of PSA-glycans represented predominantly fucosylated, partially sulfated 2,6-branched isomers of triantennary as well as tetraantennary complex type glycans, whereas carbohydrate chains bearing the HNK1-epitope were dominated by diantennary species carrying in part bisecting GlcNAc residues. Non-PSA/HNK1 glycans exhibited a highly heterogeneous pattern of partially truncated, mostly diantennary structures being characterized by the presence of additional fucose, bisecting GlcNAc and/or sulfate residues. In conclusion, our results revealed that the glycosylation pattern of murine NCAM displays high structural and regional selectivity, which might play an important role in controlling the biological activities of this molecule. PMID- 11425799 TI - Increasing the intra-Golgi pH of cultured LS174T goblet-differentiated cells mimics the decreased mucin sulfation and increased Thomsen-Friedenreich antigen (Gal beta1-3GalNac alpha-) expression seen in colon cancer. AB - Mucins in ulcerative colitis and colon cancer share common properties of reduced sulfation and increased oncofetal carbohydrate antigen expression. It has previously been shown that there is no simple correlation between these changes and the activity of the relevant glycosyl-, sialyl-, and sulfo-transferases. We examined mucin sulfation and expression of oncofetal Thomsen-Friedenreich (TF) antigen (galactosyl beta1-3N-acetylgalactosamine alpha-) in the goblet cell differentiated human colon cancer cell line LS174T following treatment with bafilomycin A(1, )which raises intra-Golgi pH, or monensin, which disrupts medial trans Golgi transport. Cells were dual-labeled with sodium [(35)S]-sulfate and D [6-(3)H(N)]-glucosamine hydrochloride, or labeled with L-[U-(14)C]-threonine alone. Mucin was purified using Sepharose CL-4B gel filtration. Mucin sulfo Lewis(a) and TF antigen expression were assessed using the F2 anti-sulfo-Lewis(a) monoclonal antibody and peanut agglutinin binding respectively. Bafilomycin (0.01 microM; 48 h) reduced total mucin sulfation, expressed relative to incorporation of glucosamine, to 0.50 +/- 0.04 d.p.m. [(35)S]-sulfate per d.p.m. [(3)H] glucosamine compared to control, 0.84 +/- 0.05 (p < 0.001, n = 16). This was accompanied by 50.3 +/- 8.0% increased expression of TF antigen (p < 0.01) and 50.1 +/- 5.5% decreased expression of sulfo-Lewis(a) (p < 0.01). The reduced sulfate:glucosamine ratio was largely due to increased incorporation of glucosamine into newly synthesized mucin rather than reduction in total sulfate incorporation. In contrast, monensin only reduced total mucin glycosylation at concentrations > 0.1 microM and had no significant effect on mucin sulfation or TF expression. Intra-Golgi alkalinization affects mucin glycosylation, resulting in decreased mucin sulfation and increased expression of TF antigen, changes that mimic those seen in cancerous and premalignant human colonic epithelium. PMID- 11425800 TI - Selective expression of different fucosylated epitopes on two distinct sets of Schistosoma mansoni cercarial O-glycans: identification of a novel core type and Lewis X structure. AB - The glycobiology of Schistosoma mansoni is dominated by developmentally regulated expression of various fucosylated structures, most notably the Lewis X epitope and a multifucosylated sequence, Fuc alpha1-->2Fuc alpha1-->, in its various forms. For the infective cercarial stage, Lewis X has been structurally identified on glycosphingolipids and N-glycans of total glycoprotein extracts, and a population of multifucosylated glycoproteins were found to carry a unique terminal sequence, +/-Fuc alpha1-->2Fuc alpha1-->[3GalNAc beta1-->4(Fuc alpha1- >2Fuc alpha1--> 2Fuc alpha1-->3) GlcNAc beta1-->3Gal alpha1-->](n), on their O glycans. Using a mass spectrometry approach coupled with chromatographic separation, sequential exoglycosidase digestion, periodate oxidation, and other chemical derivatization, we demonstrate that Lewis X could also be carried on the cercarial O-glycans, but the two distinctive sets of fucosylated epitopes were conjugated to two different core structures. Lewis X, lacNAc, or single GlcNAc was found to attach directly to the -->3Gal beta1-->3GalNAc core and indirectly via another beta-Gal residue branching off from C6 of the reducing end GalNAc to give a biantennary-like structure. The -->3(+/-Gal beta1-->6)Gal beta1-->3(- >3Gal beta1-->6)GalNAc core thus characterized represents a novel core type for O glycans. In contrast, the previously characterized multifucosylated terminal sequences were carried on conventional type 1 and 2 cores. The smallest structures of the reductively released O-glycans were defined as GalNAc beta1- >4GlcNAc beta1-->3Gal beta1-->3GalNAcitol with a total of two to four fucoses attached to the terminal lacdiNAc. alpha-Galactosylation of the nonreducing terminal beta-GalNAc instead of fucose capping leads to further elongation with another lacdiNAc unit that could also extend directly from C6 of the reducing end GalNAc and similarly elongated or terminated. PMID- 11425801 TI - Mouse ST6Gal sialyltransferase gene expression during mammary gland lactation. AB - The sialyltransferase ST6Gal mediates the biosynthetic addition of sialic acid, via an alpha2,6 linkage, to the nonreducing end of terminal lactosamine structures. Transcription of the murine ST6Gal gene, Siat1, is regulated by the selective use of multiple promoters in a tissue- and development-specific manner. Here we report that Siat1 mRNA expression is dramatically elevated in lactating (relative to virgin) mouse mammary gland. The predominant ST6Gal mRNA species expressed in lactating mammary gland is a heretofore undocumented isoform containing a unique 5'-untranslated region originating from the mouse Siat1 genetic region, now defined as Exon L, residing 549-bp 5' of the previously characterized Exon X(2). Thus, the novel ST6Gal mRNA form initiates transcription from the region designated as p4 and incorporates the unique sequence from Exon L in 5'-juxtaposition to commonly shared sequences encoded on Exon I to Exon VI. In contrast, cells derived from virgin mammary tissue expressed only the housekeeping mRNA form derived from p3, with Exon O sequence preceding Exons I VI. The Exon L-containing, p4 class of mRNA was also not detected in a survey of eight other mouse tissues. Previous reports have indicated a strong correlation between mammary cancers and elevated ST6Gal expression in rats and in human patients. However, we uncovered neither elevated expression of ST6Gal mRNA nor appearance of p4 class in mouse breast carcinomas experimentally induced by transformation with the polyoma-middle T oncogene. A number of established breast carcinoma cell lines were also examined, with ST6Gal mRNA and activity generally low. Moreover, with the exception of the Shionogi cell line, p4 class of ST6Gal mRNA was not expressed in any of the mouse breast carcinoma specimens examined. Taken together, our data indicate that murine ST6Gal induction during lactation is achieved by de novo recruitment of a normally silent promoter. Furthermore, the data provide no support for elevated Siat1 expression on the mRNA level in association with murine mammary gland carcinogenesis. With the single exception of the Shionogi cell line, the p3 class remains the predominant ST6Gal mRNA expressed in all other murine mammary carcinoma cells examined. PMID- 11425802 TI - The UDPase activity of the Kluyveromyces lactis Golgi GDPase has a role in uridine nucleotide sugar transport into Golgi vesicles. AB - In Saccharomyces cerevisiae a Golgi lumenal GDPase (ScGda1p) generates GMP, the antiporter required for entry of GDP-mannose, from the cytosol, into the Golgi lumen. Scgda1 deletion strains have severe defects in N- and O-mannosylation of proteins and glycosphingolipids. ScGda1p has also significant UDPase activity even though S. cerevisiae does not utilize uridine nucleotide sugars in its Golgi lumen. Kluyveromyces lactis, a species closely related to S. cerevisiae, transports UDP-N-acetylglucosamine into its Golgi lumen, where it is the sugar donor for terminal N-acetylglucosamine of the mannan chains. We have identified and cloned a K. lactis orthologue of ScGda1p. KlGda1p is 65% identical to ScGda1p and shares four apyrase conserved regions with other nucleoside diphosphatases. KlGda1p has UDPase activity as ScGda1p. Transport of both GDP-mannose, and UDP GlcNAc was decreased into Golgi vesicles from Klgda1 null mutants, demonstrating that KlGda1p generates both GMP and UMP required as antiporters for guanosine and uridine nucleotide sugar transport into the Golgi lumen. Membranes from Klgda1 null mutants showed inhibition of glycosyltransferases utilizing uridine- and guanosine-nucleotide sugars, presumably due to accumulation of nucleoside diphosphates because the inhibition could be relieved by addition of apyrase to the incubations. KlGDA1 and ScGDA1 restore the wild-type phenotype of the other yeast gda1 deletion mutant. Surprisingly, KlGDA1 has only a role in O glycosylation in K. lactis but also complements N-glycosylation defects in S. cerevisiae. Deletion mutants of both genes have altered cell wall stability and composition, demonstrating a broader role for the above enzymes. PMID- 11425803 TI - Neighboring cysteine residues in human fucosyltransferase VII are engaged in disulfide bridges, forming small loop structures. AB - Among alpha 3-fucosyltransferases (alpha3-FucTs) from most species, four cysteine residues appear to be highly conserved. Two of these cysteines are located at the N-terminus and two at the C-terminus of the catalytic domain. FucT VII possesses two additional cysteines in close proximity to each other located in the middle of the catalytic domain. We identified the disulfide bridges in a recombinant, soluble form of human FucT VII. Potential free cysteines were modified with a biotinylated alkylating reagent, disulfide bonds were reduced and alkylated with iodoacetamide, and the protein was digested with either trypsin or chymotrypsin, before characterization by high-performance liquid chromatography/electrospray ionization mass spectrometry. More than 98% of the amino acid sequence for the truncated enzyme (beginning at amino acid 53) was verified. Mass spectrometry analysis also demonstrated that both potential N-linked sites are occupied. All six cysteines in the FucT VII sequence were shown to be disulfide-linked. The pairing of the cysteines was determined by proteolytic cleavage of nonreduced protein and subsequent analysis by mass spectrometry. The results demonstrated that Cys(68)-Cys(76), Cys(211)-Cys(214), and Cys(318)-Cys(321) are disulfide linked. We have used this information, together with a method of fold recognition and homology modeling, using the (alpha/beta)(8)-barrel fold of Escherichia coli dihydrodipicolinate synthase as a template to propose a model for FucT VII. PMID- 11425804 TI - ART in HIV-infected couples: has the time come for a change of attitude? AB - Today, in developed countries, many HIV-infected people remain in good health thanks to antiviral medication. A growing number of them want to have children. Medical possibilities for preventing contamination of the partners of seropositive men, through assisted reproduction, and of children thanks to antiviral medicines during pregnancy, are summarized. These changes result in ethical considerations which lead the authors to question the conventional systematic medical advice against pregnancy and has encouraged them to assist reproduction for a number of these couples. Today, the balance between the importance of the message of prevention and the benefit for patients of being assisted in their desire for a child has tilted towards medical intervention. It would seem legitimate today to intervene in the most favourable situations rather than see these couples take the risk of spontaneous conception outside health care structures. This implies the need to adapt medical structures (separate laboratory, appropriate procedure, precise protocols). This approach, which is coherent from the scientific point of view, respects both the autonomy of people carrying HIV as well as the essential interests of the child, in being born uninfected, and also has the enormous advantage of allowing access to parenthood without destroying the consistency of the message of prevention of sexual contamination. PMID- 11425805 TI - Is there a place for different isoforms of FSH in clinical medicine? IV. The clinician's point of view. AB - Follicle stimulating hormone (FSH) plays an important role in gametogenesis in both men and women. In men it is required for the establishment and probably maintenance of spermatogenesis. In women it stimulates the growth of ovarian follicles and fluctuations in its secretion are involved in the selection of the ovulatory follicle. It is now possible by recombinant DNA technology to synthesize a range of isoforms of FSH with differing biological action and half life which would provide a greater degree of flexibility in the treatment of women with anovulatory infertility. Longer acting isoforms of FSH would find clinical application in the treatment of hypogonadotrophic men. PMID- 11425806 TI - Avoiding multiple pregnancies in ART: evaluation and implementation of new strategies. AB - Strategies for reducing the multiple pregnancies which increase perinatal mortality and morbidity in women undergoing IVF are discussed. Elective single embryo transfer with the promise of subsequent transfer of frozen-thawed embryos would achieve the goal of a single healthy child as a result of IVF treatment. The urgent establishment of a definitive trial comparing elective single versus double embryo transfer is advocated. PMID- 11425807 TI - Induction of delayed follicular rupture in the human by the selective COX-2 inhibitor rofecoxib: a randomized double-blind study. AB - BACKGROUND: The aims of the present study were to examine whether periovulatory administration of a cyclo-oxygenase (COX)-2 inhibitor affects human ovulation and endocrine parameters. METHODS: Thirteen healthy women, 30-40 years of age, without hormonal treatment and with regular menstrual cycles (27-34 days), were given the selective COX-2 inhibitor rofecoxib (n = 6) or placebo (n = 7) in a random double-blind fashion. In an initial control cycle, serial hormonal analyses, detection of a measurable mid-cycle urine LH peak and transvaginal ultrasound scans were performed to confirm normal ovulatory and endocrinological cyclic patterns, in all participating women. During the subsequent treatment cycle, serial ultrasound scans were performed. When the dominant follicle reached 14-16 mm in diameter, 25 mg rofecoxib or placebo was taken orally, once daily for 9 consecutive days, during which follicle size was monitored daily by ultrasound scans and serial hormone analyses were performed. RESULTS: Four of the six women who received rofecoxib demonstrated delayed follicle rupture, >48 h after the LH peak, when compared with the placebo group, who all had follicular rupture >36 h after the detected LH peak. No differences in peripheral serum concentrations of progesterone, oestradiol, LH and FSH were observed between placebo and rofecoxib groups, when analysed at specified time intervals. CONCLUSIONS: This study suggests that selective COX-2 inhibition has a negative, local effect on human ovulation, resulting in delayed follicular rupture, without affecting peripheral hormonal cyclicity. PMID- 11425808 TI - Preferential vascular-based transfer from vagina to the corpus but not to the tubal part of the uterus in postmenopausal women. AB - BACKGROUND: Vaginal administration of progesterone during infertility treatment has therapeutic advantages over oral administration. However, the reasons for this are poorly defined. To demonstrate a preferential vagina-to-uterus distribution of substances, we investigated cold distribution from vagina to the uterus and rectum. METHOD: In 10 postmenopausal women, thermoprobes were inserted into the uterine cavity and in the rectum at <9 cm or at >9 cm from the anus; temperatures were subsequently measured during 10 min flushing of vagina with cold saline. RESULTS: After 10 min, temperature decreased as follows: uterus, tubal angle: -0.22 +/- 0.07 degrees C, 10 (mean +/- SEM, n); uterus, middle cavity: -1.26 +/- 0.34 degrees C, 9; rectum, <9 cm insertion: -3.69 +/- 0.68 degrees C, 3; rectum, >9 cm insertion: -0.51 +/- 0.19 degrees C, 6. CONCLUSIONS: Despite obviously different distances to the vagina of the uterine and the low rectal probes (<9 cm) the temperature decrease occurred at the same time. Cold transfer from vagina to the uterus and rectum is probably not the result of simple diffusion but of a vascular counter-current transfer. Differential cooling of corpus and tubal angles suggests a different arterial supply; while uterine corpus is supplied from the uterine artery, the tubal angles seem to be mainly supplied from the ovarian artery via the tubal arcade. PMID- 11425809 TI - Plasma concentrations of nitrate during the menstrual cycle, ovarian stimulation and ovarian hyperstimulation syndrome. AB - BACKGROUND: Nitric oxide (NO) is predominantly a locally acting mediator, affecting several functions in the human female reproductive tract. In vivo, it is quickly metabolized to its stable end product nitrate, which is cleared by the kidney. METHODS AND RESULTS: The aim of the present study was to evaluate possible fluctuations of plasma nitrate concentrations during the menstrual cycle, ovarian stimulation as well as ovarian hyperstimulation syndrome (OHSS). During the menstrual cycle (n = 19 women) the mean nitrate concentrations were between 26.7 and 29.5 micromol/l at all stages except for the day of ovulation, when the concentrations were significantly (P < 0.001) increased (mean 37.2 micromol/l +/- 2.0). Significantly lower concentrations of plasma nitrate (P < 0.01) were measured at the end of gonadotrophin-releasing hormone (GnRH) down regulation (24.6 micromol/l +/- 1.4) compared with the concentrations found at day 8 of follicle-stimulating hormone (FSH) stimulation (34.9 micromol/l +/- 2.6) and at the day of human chorionic gonadotrophin (HCG) (35.6 micromol/l +/- 3.3). The concentrations of nitrate (33.4 micromol/l +/- 3.4) in women with OHSS (n = 13) were similar to those seen 5 days after embryo transfer (33.2 micromol/l +/- 2.3). CONCLUSIONS: The results indicate that NO synthesis is increased at the time of spontaneous ovulation. GnRH treatment inhibits NO synthesis, while NO production is not increased in women with OHSS. PMID- 11425810 TI - Leptin concentrations in hirsute women with polycystic ovary syndrome or idiopathic hirsutism: influence on LH and relationship with hormonal, metabolic, and anthropometric measurements. AB - BACKGROUND: The known association between leptin, obesity and insulin action suggests that leptin may have a role in polycystic ovarian syndrome (PCOS) but this has only been addressed peripherally. METHODS: We assessed the influence of leptin on LH and investigated the relationship between leptin and body mass index (BMI), waist:hip ratio (WHR), androgen concentrations, fasting insulin and insulin:glucose ratio (IGR) in 27 women with PCOS and in 20 age- and weight matched women with regular, ovulatory menstrual cycles and idiopathic hirsutism (IH). RESULTS: Leptin concentrations were significantly higher in obese PCOS women than in normal weight women with either PCOS or IH (P = 0.0028), but did not differ between obese women with PCOS and IH. WHR, insulin concentrations and IGR were significantly higher in obese PCOS patients in comparison with the three other groups. In IH patients, the association between leptin concentrations and WHR was lost after adjustment for BMI. In PCOS patients, a significant correlation was observed between leptin and fasting insulin concentrations, IGR, WHR and LH. After adjustment for BMI, only the correlation with LH remained significant. A stepwise regression model was set up with LH as the dependent variable to test the hypothesis that the concentrations of leptin might be modulating the concentrations of LH in PCOS patients. The relationship of LH concentrations with IGR was found to be BMI dependent. In contrast, leptin concentrations contributed negatively and significantly to LH concentrations, independently of either BMI or IGR. CONCLUSIONS: We speculate that the known attenuation in basal or stimulated response of LH in obese PCOS patients might be related to leptin resistance, which could influence LH hypersecretion. In IH ovulatory patients, normal LH concentrations suggest the presence of preserved regulatory mechanisms of GnRH pulsatility. Further studies are needed to specifically investigate the proposed correlation between leptin and GnRH modulation in PCOS. PMID- 11425811 TI - Mid-luteal serum inhibin-A concentration as a marker of endometrial differentiation. AB - BACKGROUND: Recent studies have indicated that the corpus luteum is a major source of circulating inhibin-A and serum concentrations of inhibin-A may reflect the human luteal function. The present prospective study was undertaken to determine the usefulness of mid-luteal serum concentrations of inhibin-A as markers of endometrial receptivity (as assessed by histological dating and alphavbeta3 integrin expression) and whether they are better predictors of endometrial function than serum progesterone. METHODS: Consecutive infertile women (experimental group, n = 50) with regular menstrual cycles, and fertile women who were requesting contraception and had regular menstrual patterns and normal secretory endometria (control group, n = 10) were included. In all women basal body temperature, luteal serum concentrations of oestradiol, progesterone, prolactin, and inhibin-A, and endometrial biopsies were used in the same cycle to assess luteal function. RESULTS: Out-of-phase mid-secretory endometria were detected in 17 of the 50 infertile women. Lack of alphavbeta3 integrin expression was detected in 27 of the 50 mid-luteal endometrial biopsies. Thus, hormonal concentrations were compared in the mid-luteal phase between the following eight groups of women: group 1 (n = 10), control fertile women; group 2 (n = 50), infertile women (all); subdivided into group 3 (n = 33), with in-phase biopsies; group 4 (n = 17), with out-of-phase endometria; group 5 (n = 23), expressing alphavbeta3 integrin in endometria; group 6 (n = 27), whose endometria did not express alphavbeta3 integrin; group 7 (n = 18), with both in-phase endometrial biopsy and alphavbeta3 integrin expression; and finally group 8 (n = 12), whose endometria were out-of-phase and did not express alphavbeta3 integrin. Mid-luteal serum concentrations of oestradiol, progesterone, prolactin, and inhibin-A of the seven infertile groups were similar to those of the control group of fertile women. No statistically significant difference between the infertile groups was observed for any hormonal parameter considered. CONCLUSION: Mid-luteal serum inhibin-A determination does not accurately reflect endometrial function/maturation and it is not a better indicator of endometrial luteal phase dysfunction than mid-luteal serum progesterone. PMID- 11425812 TI - Relationship between follicle size and gonadotrophin surge attenuating factor (GnSAF) bioactivity during spontaneous cycles in women. AB - BACKGROUND: We have previously demonstrated that follicles < or =11 mm diameter from women undergoing IVF contain higher concentrations of gonadotrophin surge attenuating factor (GnSAF) bioactivity than large follicles from the same ovaries. METHODS: To determine whether this finding is relevant to spontaneous cycles, follicular fluid aspirated from 37 follicles between 3 and 25 mm in diameter from 14 pairs of ovaries from regularly cycling women undergoing total abdominal hysterectomy and bilateral salpingoophorectomy for benign gynaecological disease was pooled into size categories (3 + 4, 5 + 6, 7 + 8, 9 + 10, 11 + 12, 14 + 15, 18 and 25 mm). These pools were bioassayed for GnSAF and inhibin-A, inhibin-B and activin-A concentrations were determined. RESULTS: Follicles of 5 + 6 mm diameter contained the highest concentrations of GnSAF bioactivity (reducing GnRH-induced LH secretion to 38 +/- 8% of control, P < 0.001), while those of 25 mm diameter contained one quarter of this concentration (reducing GnRH-induced LH secretion to 72 +/- 2% of control, P < 0.05). GnSAF bioactivity was closely related to follicle size (r = -0.836, P < 0.01), but not to inhibin-A, inhibin-B or activin-A concentrations. CONCLUSIONS: The finding that small follicles contain high concentrations of GnSAF bioactivity, which fall as folliculogenesis progresses during spontaneous cycles, support the hypothesis that GnSAF has a role in preventing the premature onset of the LH surge in women. PMID- 11425813 TI - Absent biologically relevant associations between serum inhibin B concentrations and characteristics of polycystic ovary syndrome in normogonadotrophic anovulatory infertility. AB - BACKGROUND: Dominant follicle selection is disturbed in normogonadotrophic anovulatory infertility [World Health Organization (WHO) 2] and remaining early antral follicles are either healthy or atretic. This study was conducted to investigate whether inhibin B serum concentrations (produced by healthy small antral follicles) represent the extent of ovarian abnormalities in WHO 2 women and patients with polycystic ovarian syndrome (PCOS), constituting a subgroup of WHO 2 patients. METHODS AND RESULTS: Ultrasonographic and endocrine characteristics in 379 WHO 2 patients and 30 normo-ovulatory controls were compared. In the WHO 2 patients, the PCOS subgroup and the controls, inhibin B concentrations were similar. Inhibin B concentrations were weakly but significantly correlated with the total number of ovarian follicles (r = 0.282; P < 0.001), LH (r = 0.347; P < 0.001), and testosterone (r = 0.269; P < 0.001) but not with serum oestradiol concentrations (r = 0.057). Most (71%) patients with elevated inhibin B also presented with increased concentrations of LH and/or hyperandrogenaemia. In a subgroup of 190 subjects, classified as PCOS based on hyperandrogenaemia and polycystic ovaries, elevated inhibin B concentrations were found in 23% of cases. Aforementioned correlations were similar in PCOS as in WHO 2 patients. CONCLUSION: In conclusion, inhibin B serum concentrations are normal in WHO 2 and PCOS women, suggesting a normal number of healthy early antral follicles despite increased overall follicle numbers in PCOS. PMID- 11425814 TI - Genetic sperm defects and consanguinity. AB - BACKGROUND: The existence of a genetic component to human infertility has been suggested, although neither the specific abnormalities involved, nor their genetic mechanism of transmission, are currently defined. We have examined, by transmission electron microscopy (TEM), ejaculate from 1600 males with fertility problems. Among the subjects studied, we focused on a group of patients whose family histories revealed different degrees of consanguinity, in order to evaluate the relationship between consanguinity and particular sperm alterations. METHODS AND RESULTS: A total of 64 consanguineous individuals were identified. In this group, excluding two azoospermic patients, 17 patients (27%) were found to have well recognized genetic ultrastructural defects affecting their entire sperm population: eight subjects had spermatozoa with "stunted tails", four "detached tail" spermatozoa, two "Kartagener's syndrome", two "miniacrosome" and one "round headed" spermatozoa. Since these alterations affect the total sperm population and do not respond to medical treatment, they are suspected of having a genetic origin. The remaining group of 1506 non-consanguineous patients suffered from the same genetic defects in only 15 cases (<1%). CONCLUSIONS: From the data presented, it appears that some very peculiar and rare sperm defects may have a genetic basis since they occur more frequently in consanguineous patients, and are related to different degrees of consanguinity. Since the ejaculate of the remaining patients, both consanguineous and not, showed diverse types of ultrastructural sperm anomalies that did not affect the entire sperm population, they might represent pathologies lacking a genetic basis. PMID- 11425815 TI - Use of fertility drugs and risk of ovarian cancer. AB - BACKGROUND: The potential association between fertility drugs and risk of ovarian cancer has been analysed using data from a case-control study conducted between January 1992 and September 1999 in four Italian areas. METHODS: Cases were 1031 women (median age 56, range 18-79 years) with incident, histologically confirmed epithelial ovarian cancer. Controls were 2411 women (median age 57, range 17-79 years) residing in the same geographical areas and admitted to the same network of hospitals for cases for a wide spectrum of acute, non neoplastic, non hormone related conditions. RESULTS: A total of 15 cases and 26 controls reported use of fertility drugs. The corresponding odds ratio (OR) was 1.3 (95% confidence interval 0.7-2.5). The OR was 1.2 for women reporting last use <25 years before interview and 1.3 for >25 years. CONCLUSIONS: Considering calendar year at use, the OR was non-significantly above unity for women reporting fertility drug use after 1970. The OR was 0.6 among nulliparous women and 1.9 among parous ones. PMID- 11425816 TI - Vascular endothelial growth factor, platelet endothelial cell adhesion molecule-1 and vascular cell adhesion molecule-1 in the follicular fluid of patients undergoing IVF. AB - BACKGROUND: The aim of our study was to measure concentrations of vascular endothelial growth factor (VEGF), platelet endothelial cell adhesion molecule-1 (PECAM-1) CD31 and vascular cell adhesion molecules (VCAM-1) in the follicular fluid of women treated with assisted reproduction technology to determine whether these proteins might be outcome markers. METHODS: Follicular fluid was collected from 75 patients < or =40 years undergoing oocyte retrieval procedures at our tertiary hospital during 1997 and 1998: 50 with tubal disease, 12 with endometriosis, and 13 whose partners had been diagnosed with severe oligoasthenoteratozoospermia. This retrospective analysis considered age and information about treatment and outcome for all these women who had undergone fewer than three assisted reproduction attempts. RESULTS: Nineteen women became pregnant (defined by human chorionic gonadotrophin concentrations and embryonic cardiac activity 1 month after follicular aspiration); 56 did not. Women did not differ significantly in their follicular fluid concentrations of VEGF, sCD31 and VCAM-1 according to cause of infertility, or assisted reproduction outcome, or age. Follicular fluid concentrations of VEGF were significantly correlated with the number of gonadotrophin ampoules administered (P < 0.012), and follicular fluid concentrations of sVCAM-1 with the fertilization rate (P < 0.01). Follicular fluid concentrations of VEGF and sVCAM-1 were also correlated (P < 0.007). CONCLUSIONS: Our results do not suggest that VEGF, CD31, or sVCAM-1 in follicular fluid predict assisted reproduction outcome, especially among patients < or =40 years old. The correlation of a high fertilization rate and sVCAM-1 in follicular fluid suggests that sVCAM-1 might be a marker of fertilization. PMID- 11425817 TI - Effects of female and male smoking on success rates of IVF and gamete intra Fallopian transfer. AB - BACKGROUND: Smoking by both male and female partners may play a significant role in the success rates of assisted reproductive technologies. The objective of this 5-year prospective study was to investigate the influence of cigarette smoking by the wife, husband, and couple at various time points (e.g. lifetime, week prior, or during the procedures) on different biological parameters of IVF and gamete intra-Fallopian transfer (GIFT). METHODS AND RESULTS: A total of 221 couples, aged >20 years, of Caucasian, Black, Asian or Hispanic descent were recruited from seven infertility clinics located in Southern California. Couples (i.e. either female or male or both) who ever smoked compared with non-smokers, had adjusted relative risks (RR) of 2.41 (95% CI 1.07-5.45, P = 0.03) of not achieving a pregnancy, and 3.76 (95% CI 1.40-10.03, P < 0.01) of not having a live birth delivery, while adjusting for potential confounders. For couples who smoked for >5 years, there was an adjusted RR = 4.27 of not achieving a pregnancy (95% CI l.53-11.97, P = 0.01). The number of oocytes retrieved decreased by 40% for couples (smokers, n = 6) and by 46% for men who smoked during the week of the visit for IVF or GIFT. Women who smoked in their lifetime had adjusted risks of 2.71 of not achieving a pregnancy (95% CI 1.37-5.35, P < 0.01), and 2.51 (95% CI 1.11-5.67, P < 0.03) of not having a live birth delivery. CONCLUSIONS: There is compelling evidence that couples should be made aware that smoking years before undergoing IVF and GIFT can impact treatment outcome. This study may also provide insight into the timing and effects of male and female smoking on natural reproduction. PMID- 11425818 TI - Friendly IVF: patient opinions. AB - BACKGROUND: The present trend towards low stimulation protocols in IVF calls for evaluation of patient attitudes. METHODS: This study compared results of a 23 item questionnaire mailed to 167 patients receiving a low stimulation type of regimen (LS-IVF) (unstimulated cycle or clomiphene) and to 116 patients treated by a standard protocol (S-IVF) (long-down regulation with gonadotrophin-releasing hormone analogue and FSH or human menopausal gonadotrophin). RESULTS: Around two thirds of all responders in both groups deemed side-effects important, but side effects and stress associated with hormone treatment were more prevalent in patients receiving S-IVF than LS-IVF. Stress due to cycle cancellation was acceptable, mild or not perceptible in significantly (P < 0.005) more patients receiving LS-IVF [48% (36/75)] compared with patients having S-IVF [26% (8/31)]. Of patients having tried the LS-IVF protocol, 93% (125/135) would suggest either LS-IVF or a sequence of this and S-IVF as a future treatment "package" compared with only 53% (33/63) in the S-IVF group (chi(2) = 43.08, P < 0.0001). The LS-IVF group showed a significant trend towards acceptance of higher number of treatment cycles. CONCLUSIONS: The patients seemed to prefer the simplicity and short duration of a low stimulation regimen in spite of drawbacks such as a high risk of cycle cancellations and accordingly the necessity for more treatment cycles. PMID- 11425819 TI - Uterine artery Doppler velocimetry and the outcome of pregnancies resulting from ICSI. AB - BACKGROUND: An increased incidence of pregnancy complications following assisted reproduction has been reported. The use of uterine artery Doppler ultrasound may aid the prediction of such complications. METHODS: Doppler was performed at 18-24 weeks gestation in 114 singleton and 32 twin pregnancies after intracytoplasmic sperm injection (ICSI) and compared with a control group matched for age, parity and plurality. Outcome variables included gestational age at delivery, prematurity, preterm premature rupture of membrane (PPROM), birth weight, birth weight discordance of >20% in twins, small for gestational age (SGA), mode of delivery, development of pre-eclampsia and placental abruption. RESULTS: Compared with the controls, there were no significant differences concerning uterine Doppler parameters, pregnancy complications and the neonatal outcome, either in singleton or in twin pregnancies. According to Doppler results and/or risk factors by medical history, 42% of singleton ICSI and 39% of spontaneous singleton pregnancies were considered as high risk. In singletons, abnormal Doppler findings were associated with pre-eclampsia in 22% and SGA in 26% of ICSI patients, compared with 33 and 21% in controls; in contrast, 0 and 10% in ICSI and 3 and 6% in controls showed these complications but no risk factors respectively. No correlation was found between PPROM, prematurity, the rate of Caesarean section and pathological Doppler results. CONCLUSIONS: Uterine Doppler examination holds the potential to identify patients with an increased risk for developing pregnancy complications. According to our results, this risk is not elevated after ICSI treatment, therefore the decision of offering an intensified antenatal care should be based on the results of Doppler examination or risks by medical history rather than the mode of conception. PMID- 11425820 TI - Fresh embryo transfer is more effective than frozen for donor oocyte recipients but not for donors. AB - BACKGROUND: Recipients of donor oocytes need to be synchronized to the donor's cycle if fresh embryos are to be transferred on the cycle of oocyte retrieval. It would be much easier to merely retrieve the oocytes from the donor, fertilize the oocytes with the recipient's male partner's spermatozoa, cryopreserve the embryos, then transfer on an oestrogen/progesterone treatment programme. METHODS: The IVF outcomes of all patients enrolled in a shared oocyte programme from January 1997 to June 1999 were reviewed. Pregnancy and implantation rates were computed and statistically analysed. RESULTS: There was a significantly higher clinical pregnancy rate for recipients who had a fresh embryo transfer compared with recipients whose first embryo transfer consisted of frozen/thawed embryos (63.4 versus 43.6%). CONCLUSIONS: Conception is more likely after fresh than frozen embryo transfer with recipients but is similar to donor conception rates. If a uterine defect, per se, even without the use of the controlled ovarian stimulation regimen, could explain the difference between fresh pregnancy and implantation rates in donors versus recipients, then these same differences would have been seen when comparing frozen transfers, but they were, in fact, similar. PMID- 11425821 TI - Effect of reduced dose of triptorelin at the start of ovarian stimulation on the outcome of IVF: a randomized study. AB - BACKGROUND: Partial pituitary desensitization using gonadotrophin-releasing hormone (GnRH) agonists may be sufficient in women undergoing controlled ovarian hyperstimulation for assisted reproduction. However, the minimal effective agonist dose remains to be determined. The aim of the study was to investigate the effect of a reduced daily dose of triptorelin, administered at the start of ovarian stimulation, on the results of IVF and intracytoplasmic sperm injection. METHODS: A total of 132 patients was randomized in two groups. Pituitary desensitization was obtained in group 1 (66 patients) with a single 3.75 mg injection (i.m.) of triptorelin. In group 2, 66 patients received 100 microg triptorelin daily, which was then reduced to 50 microg at the start of follicle stimulating hormone (FSH) stimulation. RESULTS: No significant differences were found in terms of pregnancy rate per transfer (38% in group 1 versus 34.9% in group 2), implantation rate (20.2 versus 18%) and abortion rate (8.3 versus 9.1%). The number of FSH ampoules used, as well as the number of days stimulation required, was significantly reduced in group 2 (41 +/- 26 versus 46.6 +/- 25.3, P < 0.03 and 11 +/- 1.3 versus 11.8 +/- 1.5, P < 0.002 respectively). No significant differences were seen in oestradiol concentrations and in follicle number, in the quantity of oocytes collected and fertilized, or in the number of embryos obtained or transferred. CONCLUSION: A reduced dose of triptorelin is enough for pituitary suppression during ovarian stimulation but provides no significant improvement in IVF cycle outcome when compared with depot formulation. The possibility of a shorter treatment protocol requiring lower amounts of gonadotrophins should be considered in view of its economic advantage. PMID- 11425822 TI - Training of providers in embryo transfer: what is the minimum number of transfers required for proficiency? AB - BACKGROUND: Embryo transfer represents one of the most critical procedures in the practice of assisted reproduction. The objective of this study was to identify retrospectively the minimum number of embryo transfers required to train providers properly in this skill. METHODS AND RESULTS: The study group consisted of 204 patients who received embryo transfers between January 1996 and March 2000 in a university-based programme of assisted reproduction. The main outcome measure was clinical pregnancies per embryo transfer. Five Fellow trainees performed a total of 204 embryo transfers for an overall pregnancy rate of 45.5% per embryo transfer (93/204). In comparison, the programme pregnancy rate per transfer for experienced providers was 47.3% (560/1179). A chronological graph of each individual trainee's experience for the first 50 embryo transfers performed suggested a lower initial pregnancy rate for three of the five trainees. To determine whether a learning curve might exist, results of the first 25 transfers were compared as a subgroup with the second 25 transfers. Pregnancy rates were lower for the 1-25 transfer subgroup than in the 26-50 subgroup for three of the five Fellow trainees, although the difference was not statistically significant. CONCLUSION: Clinical pregnancy rates of Fellows-in-training were indistinguishable statistically from those of experienced staff by 50 transfers. PMID- 11425823 TI - The effect of anxiety and depression on the outcome of in-vitro fertilization. AB - BACKGROUND: The study aim was to clarify the role of anxiety and depression on the outcome in assisted reproductive treatment. Previous studies on this topic have shown contradicting results, which may have been caused by population characteristics, the design of the study, or small sample sizes. METHODS: In a multicentre prospective study, 291 out of 359 (81%) consecutively invited women agreed to participate. Before down-regulation by means of gonadotrophin-releasing hormone (GnRH) analogues in a long IVF protocol, patients were asked to complete the Dutch version of the State and Trait Anxiety Inventory to measure anxiety, and the Dutch version of the Beck Depression Inventory (BDI) to measure depression. Multiple logistic regression analysis was used to analyse known predictors of pregnancy and psychological factors and their relationship with treatment outcome. RESULTS: A significant relationship was shown between baseline psychological factors and the probability to become pregnant after IVF/intracytoplasmic sperm injection (ICSI) treatment, controlling for other factors. State anxiety had a slightly stronger correlation (P = 0.01) with treatment outcome than depression (P = 0.03). CONCLUSIONS: Pre-existing psychological factors are independently related to treatment outcome in IVF/ICSI, and should therefore be taken into account in patient counselling. Psychological factors may be improved by intervention, whereas demographic and gynaecological factors cannot. Future studies should be directed towards underlying mechanisms involved and the role of evidence-based distress reduction in order to improve treatment results. PMID- 11425824 TI - Ongoing twin pregnancy after ICSI of PESA-retrieved spermatozoa into in-vitro matured oocytes: case report. AB - The recovery of immature oocytes from unstimulated ovaries followed by in-vitro maturation (IVM) is an attractive alternative to conventional IVF in the treatment of female infertility. Similarly, surgical recovery of spermatozoa from the epididymis by percutaneous sperm aspiration (PESA) has simplified the retrieval of the male gamete in treatment of men with obstructive azoospermia. We report the first ongoing clinical twin pregnancy resulting from intracytoplasmic sperm injection (ICSI) of spermatozoa retrieved by PESA into IVM oocytes. In the treatment of a 24-year old woman, 12 immature oocytes were retrieved. Six oocytes matured (maturation rate 50%) after 24-hour incubation and were inseminated by ICSI. Four oocytes had two pronuclei (fertilization rate 67%) and 3 good quality embryos were transferred. A viable twin pregnancy was confirmed by ultrasound scan. This report illustrates the use of a combination of less invasive assisted reproductive techniques in overcoming barriers to infertility. PMID- 11425825 TI - May-Grunwald-Giemsa stain for detection of spermatogenic cells in the ejaculate: a simple predictive parameter for successful testicular sperm retrieval. AB - BACKGROUND: Testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI) have become standard treatments for patients with non obstructive azoospermia. A diagnostic testicular biopsy for histopathological examination is not always predictive of TESE outcome. Moreover, it is not without potential complications. The aim of this study was to determine the value of various clinical and laboratory parameters, particularly identification of seminal spermatids using May-Grunwald-Giemsa (MGG) stain in predicting TESE results. METHODS: A total of 100 patients with non-obstructive azoospermia was subjected to clinical examination, serum FSH measurement, identification of seminal spermatids and spermatocytes using MGG staining and TESE with multiple testicular sampling. Spermatozoa were retrieved from 49% of patients. Results of TESE were compared with previous parameters in addition to histopathology. RESULTS: Testicular histopathology was, in general, an inaccurate parameter, and identification of testicular spermatids by histology predicted successful TESE in only 74% of cases. Testicular volume and serum FSH concentration also had poor predictive values. Round spermatids were identified in the ejaculate of 83.7% of TESE-positive cases, and in 22% of TESE-negative cases. CONCLUSIONS: The detection of round spermatids in semen by MGG staining provides the greatest predictive value for successful testicular sperm retrieval, and also has the advantages of simplicity, low cost and availability. PMID- 11425826 TI - High sperm aneuploidy rate in unselected infertile patients and its relationship with intracytoplasmic sperm injection outcome. AB - BACKGROUND: Men with oligoasthenoteratozoospermia (OAT) frequently undergo intracytoplasmic sperm injection (ICSI) as a treatment for their infertility. However, there is an increased risk of transmitting chromosomal abnormalities to the offspring given that natural selection is bypassed by the use of this technique and patients have an increased rate of sperm aneuploidy which, in addition, may negatively affect ICSI outcome. For this reason, the rate of sperm aneuploidy in unselected patients undergoing ICSI and its impact on ICSI performance have been evaluated. METHODS: Aneuploidy and diploidy were evaluated in spermatozoa separated by swim-up for oocyte injection, using DNA probes for chromosomes 8, 12, 18, X and Y. RESULTS: ICSI patients had sperm aneuploidy and diploidy rates significantly higher than those of 13 normozoospermic men who served as controls. Although the total aneuploidy rate varied considerably between the 18 patients, 15 of them (83%) had values above the upper range of the control group. Eighteen ICSI cycles were performed with an overall fertilization rate of 95% and a pregnancy rate of 39%. The aneuploidy rate of the 11 patients whose wives did not achieve pregnancy was slightly higher than that of pregnant couples, but the difference did not reach statistical significance. However, 10 patients in this group (91%) had a sperm aneuploidy rate well above the upper limit of the controls as compared with two patients in the "pregnant" group (29%). CONCLUSIONS: This study has shown that unselected patients undergoing ICSI had an elevated sperm aneuploidy rate. Lack of pregnancy was associated with a tendency towards an increased aneuploidy rate; however, pregnancy occurred even in the presence of an elevated sperm aneuploidy rate. PMID- 11425827 TI - Ultrastructural analysis of chromatin defects in testicular spermatids in azoospermic men submitted to TESE-ICSI. AB - BACKGROUND: Testicular sperm extraction (TESE) combined with intracytoplasmic sperm injection (ICSI) is offered to treat obstructive and non-obstructive azoospermia, but factors that influence the outcome of ICSI are not well defined. METHODS AND RESULTS: The percentage of elongated spermatids with normal chromatin condensation in azoospermic patients submitted for TESE-ICSI was determined. The quantitative analysis could be applied to nine of 19 biopsies classified as incomplete late maturation arrest (LMA) and compared with 10 biopsies with normal spermatogenesis. The percentage of elongated spermatids with normal chromatin was lower in LMA than in normal histology (mean 4.4%, range 0-20, and mean 52.9%, range 40-70 respectively; P = 0.0001). The percentage of elongated spermatids with normal chromatin was negatively correlated with the serum concentration of FSH (r = -0.86, P < 0.0001) and the number of degenerated germ cells per 100 Sertoli cells nuclei (r = -0.68; P < 0.0001), while it was positively correlated with the number of elongating spermatids per 100 Sertoli cell nuclei (r = 0.81; P < 0.0001). The percentage of elongated spermatids with normal chromatin was not correlated with the rate of oocyte fertilization, while the delivery rate/cycle was higher in cases with normal histology compared with cases of LMA. CONCLUSIONS: These preliminary data suggest that an altered chromatin condensation is a ubiquitous defect in spermatids of non-obstructed azoospermic men submitted for TESE-ICSI. PMID- 11425828 TI - Differential gene expression in pre-implantation embryos from mouse oocytes injected with round spermatids or spermatozoa. AB - BACKGROUND: The use of immature male germ cells to fertilize human oocytes raises several questions. Spermatozoa are normally quiescent, but many genes are transcribed post-meiotically in round spermatids. This creates a novel situation for the oocyte. We have therefore explored the effects on early embryonic development of introducing a fully transcriptionally active round spermatid into the oocyte. METHODS AND RESULTS: Following the micro-injection of spermatozoa or spermatids into mouse oocytes we have analysed the expression, at various times, of six genes in the resulting embryo. Spermatozoa and spermatids produced similar fertilization rates. Hprt was expressed in all embryos at all stages tested. Hsp70.1 was found normally during the 2-cell stage and repressed by the 4-cell stage in embryos from both spermatozoa and round spermatids. However, the amplitude of the signal was greatly reduced in 2-cell embryos from round spermatids. Smcy also showed a disturbed pattern of expression in embryos from round spermatids. Protamine 2, which is normally restricted to the spermatid stage, was expressed following fertilization with round spermatids, but was already repressed at the two pronuclei stage. Ube1Y, which is normally expressed post-meiotically and not during the post-implantatory development, was expressed up to the 2-cell stage in embryos from round spermatids only, and then repressed. Ube1X was also expressed up to the 2-cell stage, but in both embryo types. CONCLUSIONS: We therefore suspect that in embryos fertilized with round spermatids, regulatory mechanisms for inhibiting the inappropriate transcription of male post-meiotically expressed genes are activated following fertilization, permitting the zygotic genome activation to occur, though with some disturbances. PMID- 11425829 TI - Embryotoxicity of magainin-2-amide and its enhancement by cyclodextrin, albumin, hydrogen peroxide and acidification. AB - BACKGROUND: The channel-forming antimicrobial peptide, magainin-2-amide, interacts preferentially with negatively charged, non cholesterol-containing membranes, including those of sperm, oocytes and cells of pre-implantation embryos. Cyclodextrin and albumin remove membrane cholesterol and together with hydrogen peroxide (H2O2) are potential enhancers of embryotoxicity. METHODS: Two cell murine embryos were cultured in vitro with magainin-2-amide at a high effective concentration (250 microg/ml) and at subthreshold concentrations (166 and 200 microg/ml). Embryos treated with sub-threshold concentrations of magainin were additionally treated with cyclodextrin, bovine serum albumin or H2O2 or were cultured under acidified conditions. Cell viability was verified with propidium iodide and fluorescein diacetate. RESULTS: The embryotoxic effect of magainin and H2O2 was dose- and time-dependent. Cyclodextrin, H2O2, acidification of the medium, and to a lesser extent albumin, enhanced the embryotoxicity of magainin at sub-threshold concentrations. CONCLUSION: Magainin on its own is highly embryotoxic. Its embryotoxicity is enhanced by cyclodextrin, albumin, H2O2 and acidification. Thus, magainin which has antibacterial, antifungal and antiprotozoal activity may also have a potential role as a contraceptive agent. The harmful effects of various concentrations of the exogenous H2O2 on 2-cell stage mouse embryos are reported here, to the best of our knowledge, for the first time. PMID- 11425830 TI - Developmental ability of human oocytes with or without birefringent spindles imaged by Polscope before insemination. AB - BACKGROUND: Birefringent spindles imaged with the Polscope can predict fertilization rates after intracytoplasmic sperm injection (ICSI). The present study examined the development of human oocytes with or without birefringent spindles, imaged with the Polscope before ICSI. METHODS: Oocytes were obtained from stimulated ovaries of consenting patients undergoing oocyte retrieval for ICSI. Spindles were imaged with the Polscope combined with a computerized image analysis system. After imaging and ICSI, oocytes with or without spindles were cultured separately for examination of fertilization and embryo development. A total of 1544 oocytes from 136 cycles were examined with the Polscope and inseminated by ICSI. RESULTS: Spindles were imaged in 82% of oocytes. After ICSI, more oocytes (P < 0.05) with spindles (69.4%) fertilized normally, forming 2 pronuclei, than oocytes without spindles (62.9%). At day 3, more oocytes (P < 0.01) with spindles (66.3%) developed to 4-11 cell stages than oocytes without spindles (55.4%). Significantly more (P < 0.001) oocytes with spindles developed to morula and blastocyst by day 5 (51.1 versus 30.3%) and day 6 (53.2 versus 29.3%) compared with oocytes without spindles. CONCLUSIONS: The results indicate that the presence of a birefringent spindle in human oocytes can predict not only higher fertilization rate, but also higher embryo developmental competence. PMID- 11425831 TI - Pregnancy after cytoplasmic transfer in a couple suffering from idiopathic infertility: case report. AB - Couples suffering from idiopathic infertility are among the most difficult to treat in IVF cycles due to the unknown cause of failure to reproduce. This can lead to years of unsuccessful attempts to conceive and often the abandonment of treatment. In some couples, the only observable problem is the development of poor quality embryos. In this case report, we describe the successful use of cytoplasmic donation in a couple where the high level of embryo fragmentation and poor embryo development was thought to be the unique cause of failure to conceive after intracytoplasmic sperm injection (ICSI). We noted a strong reduction in embryo fragmentation and a consequent increase in embryo quality after the treatment. Transfer of four of the embryos receiving donor cytoplasm led to a pregnancy with two gestational sacs and the birth of healthy twins. PMID- 11425832 TI - Morbidity of 10 110 hysterectomies by type of approach. AB - BACKGROUND: Since the late 1980s, the option of laparoscopic hysterectomy has raised questions about the most suitable approach to hysterectomy. METHODS: To evaluate the influence of the type of approach, in causing or avoiding certain complaints in hysterectomies a prospective nationwide study was conducted comprising all hysterectomies for benign disease performed in Finland during 1996. The primary outcomes of interest were the operation-related morbidity, common surgical details and post-operative complications. RESULTS: A total of 10 110 hysterectomies, including 5875 abdominal, 1801 vaginal and 2434 laparoscopic operations showed a low rate of overall complications, 17.2, 23.3 and 19.0% respectively. Infections were the most common complications with incidences of 10.5, 13.0 and 9.0% in the abdominal, vaginal and laparoscopic group respectively. The most severe type of haemorrhagic events occurred in 2.1, 3.1 and 2.7% in the abdominal, vaginal and laparoscopic group respectively. Ureter injuries were predominant in laparoscopic group [relative risk (RR) 7.2 compared with abdominal] whereas bowel injuries were most common in vaginal group (RR 2.5 compared with abdominal). Surgeons who had performed >30 laparoscopic hysterectomies had a significantly lower incidence of ureter and bladder injuries (0.5 and 0.8% respectively) than those who had performed < or =30 operations (2.2 and 2.0% respectively). A decreasing trend of bowel complications was also seen with increasing experience in vaginal hysterectomies. CONCLUSIONS: This large scale observational study on hysterectomies provides novel information on operation-related morbidity of abdominal, vaginal or laparoscopic approach. The results support the importance of the experience of the surgeon in reducing severe complications, especially in laparoscopic and vaginal hysterectomies. PMID- 11425833 TI - Polycystic ovarian syndrome: a follow-up study on fertility and menstrual pattern in 149 patients 15-25 years after ovarian wedge resection. AB - BACKGROUND: The aim of this study was to evaluate fertility and menstrual pattern in women with polycystic ovarian syndrome (PCOS) 15-25 years after ovarian wedge resection (OWR). METHODS AND RESULTS: The diagnosis was based on the combination of ovarian pathology and symptoms. The 149 patients, all primarily treated at a university teaching hospital, were studied three times by means of a questionnaire up to 25 years after surgery. Kaplan-Meier analysis showed a cumulative rate of spontaneous pregnancies of 76%, increasing to 88% when induced pregnancies were included. The cumulated live birth rate was 78%. A bootstrap simulation indicated that 69.5% would develop post-operative adhesions, which could impede pregnancy in 13.4%. In the majority of the patients a regular menstrual pattern was restored up to 25 years after OWR. CONCLUSION: The results of OWR in PCOS are favourable to most modern treatments. Laparoscopic electrocautery of the ovaries is the only method equally successful, and, by being less invasive, it has made OWR history in the treatment of PCOS. PMID- 11425834 TI - The use of misoprostol prior to hysteroscopy in postmenopausal women. AB - BACKGROUND: This study examined whether oral misoprostol exerted a cervical priming effect in postmenopausal women prior to hysteroscopy. METHOD: Thirty seven patients were randomized to receive either oral misoprostol (400 microg) or placebo (vitamin B(6)) 12 h prior to hysteroscopy. The resistance of the cervix to dilatation was objectively assessed by a cervical tonometer. RESULTS: The mean baseline cervical dilatation (4.2 mm in misoprostol group versus 4.4 mm in placebo group) was similar between the two groups. The mean cumulative force measured (27.7 N in misoprostol group versus 21.8 N in placebo group) was also comparable. None of the patients suffered from any significant side-effects. CONCLUSIONS: These data showed that there were no significant benefits from giving misoprostol pre-operatively in postmenopausal women, and it was concluded that oral misoprostol had no significant cervical priming effect in postmenopausal women. PMID- 11425835 TI - Hysteroscopic resection of submucosal myomas in patients with infertility. AB - BACKGROUND: Submucosal myomas are associated with infertility, and may be treated by hysteroscopic resection. Improvement of the menstrual pattern after surgery has been examined and compared with pregnancy rate in a group of menorrhagic women with primary (n = 24) and secondary (n = 35) infertility. METHODS: The entire patient group (n = 59) was managed between January 1990 and September 1998. The submucosal myomas were intracavitary (n = 15), intramural class 1 (n = 34) and intramural class 2 (n = 10), and none was multiple submucosal myoma. The mean (+/- SD) age was 36.6 +/- 4.6 years; mean myoma size was 24.5 +/- 13.3 mm; mean duration of the procedure was 40 +/- 23 min; and mean follow-up was 26 +/- 18.8 months. Thirty-five patients had one or more associated infertility factors. RESULTS: An improvement in clinical symptoms was observed in 62% of patients. Sixteen patients (27%) conceived, and of these only six (10%) delivered at term. The pregnancy rate was significantly better when myoma was the exclusive aetiology of infertility (41.6%), and when the lesion was >/=50 mm in size (57.1%). CONCLUSIONS: Hysteroscopic myomectomy appears safe, and is effective in the control of menstrual disorders. However, the effect on infertility seems limited, particularly in terms of delivery rate. The advanced age of the patients may partly explain these results. PMID- 11425836 TI - Vaginal misoprostol as medical treatment for first trimester spontaneous miscarriage. AB - BACKGROUND: Misoprostol is effective for cervical priming prior to suction evacuation in first trimester pregnancy termination. This is the first randomized study to compare vaginal misoprostol versus expectant treatment in women presenting with spontaneous miscarriage. METHODS: Sixty women presenting with spontaneous miscarriage were recruited to the study at the Queen Mary Hospital between 1998 and 1999. They were randomized to group 1: misoprostol; and group 2: expectant management. Women in the misoprostol group received vaginal misoprostol 400 microg on days 1, 3 and 5. The expectant group was followed up according to the same schedule. Suction evacuation was performed if there was excessive bleeding or abdominal pain; or if a gestational sac was detected by transvaginal scan on day 15. RESULTS: Fifty-nine women completed the trial. Those who did not require suction evacuation up to the time of return of normal menstruation were considered to be successful. The incidence of side-effects was comparable between the two groups. Three women in the expectant group and one in the misoprostol group underwent emergency suction evacuation because of excessive bleeding. The mean duration of vaginal bleeding was similar for both groups (14.6 days in the misoprostol group versus 15.0 days in the expectant group). The successful rate in the misoprostol group was significantly higher than that of the expectant group (83.3 versus 48.3%, P < 0.05). CONCLUSION: We recommend repeated vaginal misoprostol 400 microg given on days 1, 3 and 5 as a treatment option for women with first trimester spontaneous miscarriage. PMID- 11425837 TI - Use of time to event analysis to estimate the normal duration of human pregnancy. AB - BACKGROUND: Current estimates of the average duration of human pregnancy are flawed by inaccurate estimation of the time of conception and by failure to account adequately for the effect of routine elective delivery post-term. METHODS: In this study, 1514 healthy pregnant women were studied in whom the discrepancy between the menstrual history and first trimester crown-rump length estimated gestational age was within -1 to +1 day difference. The duration of gestation was estimated using time to event analysis: non-elective delivery was taken to be the event, and elective delivery was taken to be censoring. RESULTS: The median time to non-elective delivery using the Kaplan-Meier product limit estimate was 283 days after last menstrual period (LMP) and there was no difference comparing male and female fetuses. The median was significantly greater for nulliparous women compared with multiparous women (284 versus 282 days, P < 0.0001). Multivariate analysis using Cox's proportional hazards model confirmed the independent effect of nulliparity on duration of pregnancy [hazard ratio, 0.75; 95% confidence interval (CI) 0.67-0.85] and demonstrated no effect of maternal age, previous abortions, fetal sex, high parity, or bleeding before 24 completed weeks of gestation. Bleeding in the third trimester of pregnancy was, however, associated with an earlier onset of spontaneous labour (hazard ratio, 1.38; 95% CI 1.03-1.84). CONCLUSION: This study provides a basis for predicting the probability of labour at a given gestational age at term. PMID- 11425838 TI - First-trimester screening for trisomy 21 in singleton pregnancies achieved by assisted reproduction. AB - BACKGROUND: The possible effect of assisted reproduction on first-trimester screening for trisomy 21 was examined by fetal nuchal translucency thickness (NT), maternal serum free beta-human chorionic gonadotrophin (HCG) and pregnancy associated plasma protein-A (PAPP-A). METHODS: Parameters were measured at 11-14 weeks in 411 singleton pregnancies achieved by controlled ovarian stimulation, including 220 that had undergone IVF. Results were compared with 1233 singleton pregnancies conceived spontaneously. RESULTS: In the IVF pregnancies, the median fetal NT was not significantly different from that in controls, whilst the median free beta-HCG was significantly increased, and PAPP-A was significantly decreased. In the intracytoplasmic sperm injection group, fetal NT and free beta HCG values were not significantly different from those in controls, but PAPP-A was significantly decreased. In those pregnancies achieved by ovarian stimulation, neither fetal NT, free beta-HCG nor PAPP-A were significantly different from the control group. CONCLUSIONS: In IVF pregnancies, screening for trisomy 21 by fetal NT, maternal serum free beta-HCG and PAPP-A levels may be associated with a 1.2% higher false-positive rate than in natural conception. PMID- 11425839 TI - Stress and immune mediators in miscarriage. AB - BACKGROUND: Stress is thought to be abortogenic and psycho-neuro-immunological pathways have been suggested to be involved in triggering miscarriages. From experiments in pregnant mice exposed to stress some insights into the underlying mechanisms have been gained, delineating immunological imbalances as a cause of pregnancy failure. In order to test the validity of the conclusions drawn from murine experiments and the role of stress in human pregnancy loss, the following study was performed. METHODS: We used an established perceived stress questionnaire and measured the stress score of women with a confirmed diagnosis of first trimester spontaneous abortion (n = 94). Decidual tissue was investigated by immunohistochemistry and in-situ hybridization to detect the presence and distribution of immunocompetent decidual cells [CD56(+) natural killer (NK) cells, CD8(+)and CD3(+) T cells, tryptase(+) mast cells (MCT(+)) and tumour necrosis factor (TNF)-alpha(+) cells]. The patient cohort was divided into women experiencing low or high levels of stress. RESULTS: In the decidua of women with high stress scores we observed significantly higher numbers of MCT(+), CD8(+) T cells and TNF-alpha(+) cells per mm(2) tissue (P < or = 0.05). No significant differences between individuals with lower or higher stress scores could be observed with respect to decidual CD56(+) NK and CD3(+) T cells. CONCLUSIONS: Using a questionnaire to score perceived stress in humans may be a valid approach to assess non-biased stress scores. Stress-triggered abortion in humans, identified by a questionnaire, can be linked to immunological imbalances. PMID- 11425840 TI - The seasonality of live birth is strongly influenced by socio-demographic factors. AB - The number of births varies markedly by season, but the causes of this variation are not well understood. The proposed explanations include temperature or photoperiod (affecting hormonal concentrations, sperm quality or sexual activity), seasonal variation in pregnancy loss, or cultural factors. In this paper we examined whether birth seasonality is influenced by socio-demographic factors. We used data on all live births registered in the Czech Republic in 1989 1991 (n = 387 496). Differences in the degree of seasonality between socio demographic groups (defined by maternal age, marital status, education and birth order) were examined by inspection of curves, by comparing coefficients of variations of monthly numbers of births, and by calculating the ratios of the number of births in the 3 peak months (March to May) to the number of births in the 3 lowest months (October to December). We found large differences in the size of the seasonal variation in births by socio-demographic factors. The seasonal variation was highly pronounced in mothers who were 25-34 years old, had higher education, were married, and were pregnant with their second or third child. By contrast, birth seasonality was weak in mothers who were < or =19 years or > or =35 years old, unmarried, had low education, and expected their first or fourth or higher order birth. In a multivariate model, all four socio-demographic variables contributed significantly to seasonal variation. These results suggest that the seasonality of births is, at least in this population, strongly influenced by socio-demographic factors. PMID- 11425841 TI - Social determinants of human reproduction. AB - Developed countries have experienced both some population growth and unprecedented declines in fertility rates during the last half of the twentieth century. Couples now have fewer than two children on average in most European countries and they tend to postpone these births until a later age. A decline in male fertility has been suggested by some studies of semen quality, but there is contrasting evidence of shorter times to pregnancy for couples trying to conceive. An important economic factor is the income of young men relative to their parents' incomes, which determines how they rate the ability of their own earnings to support a family. Lower relative income in the 1970s was associated with a lower fertility rate. The decline in fertility in the USA may have been attenuated by the sharp rise in female income during the late 1960s and early 1970s, allowing women to take advantage of purchased child care, thus maintaining the relative family income. The level of demand for children does not appear to be set by known psychological factors, although explanations for the desire to reproduce have been sought in biological, psychoanalytical and socio-cultural research. Recent studies indicate that adults with secure attachment relationships are more interested in being parents. Possible epidemiological factors include age at first marriage, but in Eastern Europe, where age at first marriage is as low as 22 years, fecundity rates do not exceed 1.5. When mothers' age cohorts are analysed, the mean fecundity rate has been falling since the 1920s. Health factors affecting population trends include the change in contraceptive prevalence over the last 40 years. The prevalence of sub-fertility remains close to 10%, and studies from a number of countries indicate that approximately 50% of infertile couples make use of infertility services including IVF and intracytoplasmic sperm injection which are available in 45 countries covering 78% of the world's population. It is estimated that the level of service is sufficient for less than one-third of the need. PMID- 11425842 TI - Ovarian and endometrial function during hormonal contraception. AB - This report addresses the balance of benefits and risks from changes in ovarian and endometrial function from hormonal contraception. The main mode of action of hormonal contraception is inhibition of ovulation, due chiefly to the dose of oestrogen in combined oral contraceptives. With 20 microg dosages of ethinyl oestradiol follicular activity is more common so that contraception depends on suppression of the LH surge or disruption of the endometrial cycle. In polycystic ovary syndrome (PCOS) treated with oral contraceptives, cysts become smaller and in time the ovarian volume is reduced, ovarian testosterone secretion is reduced and there are potentially favourable effects on carbohydrate and lipid metabolism. Typical oral contraceptive users in the 1980s had a lower incidence of ovarian cysts, but modern oral contraceptives do not appear to affect the incidence of functional cysts or benign epithelial cysts. Moreover, randomized controlled trials indicate that oral contraception prescriptions are unlikely to prevent the development of functional cysts or to hasten their disappearance. Oral contraceptives, however, greatly reduce pelvic pain in women with symptomatic endometriosis and improve the health-related quality of life. Bleeding is a common response with all types of hormonal contraception, but current methodology is inadequate to make accurate comparisons of different products or of different phasic formulations. With continuing use, however, combined oral contraception is associated with endometrial atrophy, the biological plausibility for a reduced risk of endometrial carcinoma. With progestin-only contraception, a number of endometrial changes are considered as possible mechanisms of the associated bleeding but it remains largely unexplained. Oral contraceptives are frequently used for treatment of dysfunctional uterine bleeding, although only one trial has been reported. Oral contraceptive use confers protection from endometrial [relative risk (RR) 0.5] and ovarian (RR 0.4) cancers and in both cases, the protection lasts for up to 2 decades after stopping use. PMID- 11425843 TI - Obstetric outcome in women with polycystic ovarian syndrome. PMID- 11425844 TI - Augmentation of ovarian response by dehydroepiandrosterone. PMID- 11425848 TI - Indirubin and indigo are potent aryl hydrocarbon receptor ligands present in human urine. AB - Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that regulates genes involved in xenobiotic metabolism, cellular proliferation, and differentiation. Numerous xenobiotic and biological compounds are known to interact with AhR, but it remains an orphan receptor, because its physiological ligand is unknown. We identified AhR ligands in human urine using a yeast AhR signaling assay and then characterized their properties. Two ligands, indirubin and indigo, were both present at average concentrations of approximately 0.2 nm in the urine of normal donors. Indirubin was also detected in fetal bovine serum and contributed half of the total AhR ligand activity. The activities of indirubin and indigo were comparable with or more potent than that of the archetypal ligand, 2,3,7,8-tetrachlorodibenzo-p-dioxin, in yeast AhR activation assays. We suggest that the endogenous levels and potencies of indirubin and indigo are such that they activate AhR-mediated signaling mechanisms in vivo. PMID- 11425849 TI - Hydrophobic ligand binding by Zn-alpha 2-glycoprotein, a soluble fat-depleting factor related to major histocompatibility complex proteins. AB - Zn-alpha(2)-glycoprotein (ZAG) is a member of the major histocompatibility complex (MHC) class I family of proteins and is identical in amino acid sequence to a tumor-derived lipid-mobilizing factor associated with cachexia in cancer patients. ZAG is present in plasma and other body fluids, and its natural function, like leptin's, probably lies in lipid store homeostasis. X-ray crystallography has revealed an open groove between the helices of ZAG's alpha(1) and alpha(2) domains, containing an unidentified small ligand in a position similar to that of peptides in MHC proteins (Sanchez, L. M., Chirino, A. J., and Bjorkman, P. J. (1999) Science 283, 1914-1919). Here we show, using serum-derived and bacterial recombinant protein, that ZAG binds the fluorophore-tagged fatty acid 11-(dansylamino)undecanoic acid (DAUDA) and, by competition, natural fatty acids such as arachidonic, linolenic, eicosapentaenoic, and docosahexaenoic acids. Other MHC class I-related proteins (FcRn, HFE, HLA-Cw*0702) showed no such evidence of binding. Fluorescence and isothermal calorimetry analysis showed that ZAG binds DAUDA with K(d) in the micromolar range, and differential scanning calorimetry showed that ligand binding increases the thermal stability of the protein. Addition of fatty acids to ZAG alters its intrinsic (tryptophan) fluorescence emission spectrum, providing a strong indication that ligand binds in the expected position close to a cluster of exposed tryptophan side chains in the groove. This study therefore shows that ZAG binds small hydrophobic ligands, that the natural ligand may be a polyunsaturated fatty acid, and provides a fluorescence-based method for investigating ZAG-ligand interactions. PMID- 11425850 TI - The human type 2 iodothyronine deiodinase is a selenoprotein highly expressed in a mesothelioma cell line. AB - Types 1 and 3 iodothyronine deiodinases are known to be selenocysteine-containing enzymes. Although a putative human type 2 iodothyronine deiodinase (D2) gene (hDio2) encoding a similar selenoprotein has been identified, basal D2 activity is not selenium (Se)-dependent nor has D2 been labeled with (75)Se. A human mesothelioma cell line (MSTO-211H) has recently been shown to have approximately 40-fold higher levels of hDio2 mRNA than mesothelial cells. Mesothelioma cell lysates activate thyroxine (T(4)) to 3,5,3'-triiodothyronine with typical characteristics of D2 such as low K(m) (T(4)), 1.3 nm, resistance to propylthiouracil, and a short half-life ( approximately 30 min). D2 activity is approximately 30-fold higher in Se-supplemented than in Se-depleted medium. An antiserum prepared against a peptide deduced from the Dio2 mRNA sequence precipitates a (75)Se protein of the predicted 31-kDa size from (75)Se-labeled mesothelioma cells. Bromoadenosine 3'5' cyclic monophosphate increases D2 activity and (75)Se-p31 approximately 2.5-fold whereas substrate (T(4)) reduces both D2 activity and (75)Se-p31 approximately 2-3-fold. MG132 or lactacystin (10 microm), inhibitors of the proteasome pathway by which D2 is degraded, increase both D2 activity and (75)Se-p31 3-4-fold and prevent the loss of D2 activity during cycloheximide or substrate (T(4)) exposure. Immunocytochemical studies with affinity-purified anti-hD2 antibody show a Se-dependent increase in immunofluorescence. Thus, human D2 is encoded by hDio2 and is a member of the selenodeiodinase family accounting for its highly catalytic efficiency in T(4) activation. PMID- 11425851 TI - New roles for the Snp1 and Exo84 proteins in yeast pre-mRNA splicing. AB - The mammalian 70K protein, a component of the U1 small nuclear ribonucleoprotein involved in pre-mRNA splicing, interacts with a number of proteins important for regulating constitutive and alternative splicing. Similar proteins that interact with the yeast homolog of the 70K protein, Snp1p, have yet to be identified. We used the two-hybrid system to find four U1-Snp1 associating (Usa) proteins. Two of these proteins physically associate with Snp1p as assayed by coimmunoprecipitation. One is Prp8p, a known, essential spliceosomal component. This interaction suggests some novel functions for Snp1p and the U1 small nuclear ribonucleoprotein late in spliceosome development. The other, Exo84p, is a conserved subunit of the exocyst, an eight-protein complex functioning in secretion. We show here that Exo84p is also involved in pre-mRNA splicing. A temperature-sensitive exo84 mutation caused increased ratios of pre-mRNA to mRNA for the Rpl30 and actin transcripts in cells incubated at the non-permissive temperature. The mutation also led to a defect in splicing and prespliceosome formation in vitro; an indication that Exo84p has a direct role in splicing. The results elucidate a surprising link between splicing and secretion. PMID- 11425852 TI - Protein tyrosine nitration in cytokine-activated murine macrophages. Involvement of a peroxidase/nitrite pathway rather than peroxynitrite. AB - Peroxynitrite, formed in a rapid reaction of nitric oxide (NO) and superoxide anion radical (O(2)), is thought to mediate protein tyrosine nitration in various inflammatory and infectious diseases. However, a recent in vitro study indicated that peroxynitrite exhibits poor nitrating efficiency at biologically relevant steady-state concentrations (Pfeiffer, S., Schmidt, K., and Mayer, B. (2000) J. Biol. Chem. 275, 6346-6352). To investigate the molecular mechanism of protein tyrosine nitration in intact cells, murine RAW 264.7 macrophages were activated with immunological stimuli, causing inducible NO synthase expression (interferon gamma in combination with either lipopolysaccharide or zymosan A), followed by the determination of protein-bound 3-nitrotyrosine levels and release of potential triggers of nitration (NO, O(2)*, H(2)O(2), peroxynitrite, and nitrite). Levels of 3-nitrotyrosine started to increase at 16-18 h and exhibited a maximum at 20-24 h post-stimulation. Formation of O(2) was maximal at 1-5 h and decreased to base line 5 h after stimulation. Release of NO peaked at approximately 6 and approximately 9 h after stimulation with interferon gamma/lipopolysaccharide and interferon-gamma/zymosan A, respectively, followed by a rapid decline to base line within the next 4 h. NO formation resulted in accumulation of nitrite, which leveled off at about 50 microm 15 h post stimulation. Significant release of peroxynitrite was detectable only upon treatment of cytokine-activated cells with phorbol 12-myristate-13-acetate, which led to a 2.2-fold increase in dihydrorhodamine oxidation without significantly increasing the levels of 3-nitrotyrosine. Tyrosine nitration was inhibited by azide and catalase and mimicked by incubation of unstimulated cells with nitrite. Together with the striking discrepancy in the time course of NO/O(2) release versus 3-nitrotyrosine formation, these results suggest that protein tyrosine nitration in activated macrophages is caused by a nitrite-dependent peroxidase reaction rather than peroxynitrite. PMID- 11425853 TI - Intracellular stability of anti-caspase-3 intrabodies determines efficacy in retargeting the antigen. AB - Although intracellular antibodies (intrabodies) are being explored as putative therapeutic and research reagents, little is known about the principles that dictate the efficacy of these molecules. In our efforts to address this issue, we generated a panel of five intrabodies, directed against catalytically inactive murine caspase-3, by screening single-chain antibody (Fv) phage display libraries. Here we determined criteria that single-chain Fv fragments must fulfill to act as efficient intrabodies. The affinities of these intrabodies, as measured by surface plasmon resonance, varied approximately 5-fold (50-250 nm). Despite their substantial sequence similarity, only two of the five intrabodies were able to significantly accumulate intracellularly. These disparities in intracellular expression levels were reflected by differences in the stability of the purified protein species when analyzed by urea denaturation studies. We observed varied efficiencies in retargeting the antigen murine caspase-3, from the cytosol to the nucleus, mediated by intrabodies tagged with an SV40 nuclear localization signal. Our results demonstrate that the intrinsic stability of the intrabody, rather than its affinity for the antigen, dictates its intracellular efficacy. PMID- 11425854 TI - Functional interaction between SEL-10, an F-box protein, and the nuclear form of activated Notch1 receptor. AB - The Notch signaling pathway is essential in many cell fate decisions in invertebrates as well as in vertebrates. After ligand binding, a two-step proteolytic cleavage releases the intracellular part of the receptor which translocates to the nucleus and acts as a transcriptional activator. Although Notch-induced transcription of genes has been reported extensively, its endogenous nuclear form has been seldom visualized. We report that the nuclear intracellular domain of Notch1 is stabilized by proteasome inhibitors and is a substrate for polyubiquitination in vitro. SEL-10, an F-box protein of the Cdc4 family, was isolated in a genetic screen for Lin12/Notch-negative regulators in Caenorhabditis elegans. We isolated human and murine counterparts of SEL-10 and investigated the role of a dominant-negative form of this protein, deleted of the F-box, on Notch1 stability and activity. This molecule could stabilize intracellular Notch1 and enhance its transcriptional activity but had no effect on inactive membrane-anchored forms of the receptor. We then demonstrated that SEL-10 specifically interacts with nuclear forms of Notch1 and that this interaction requires a phosphorylation event. Taken together, these data suggest that SEL-10 is involved in shutting off Notch signaling by ubiquitin-proteasome mediated degradation of the active transcriptional factor after a nuclear phosphorylation event. PMID- 11425855 TI - Hsp90 ensures the transition from the early Ca2+-dependent to the late phosphorylation-dependent activation of the endothelial nitric-oxide synthase in vascular endothelial growth factor-exposed endothelial cells. AB - Vascular endothelial growth factor (VEGF) exerts its angiogenic effects partly through the activation of endothelial nitric-oxide synthase (eNOS). Association with heat shock protein 90 (hsp90) and phosphorylation by Akt were recently shown to separately activate eNOS upon VEGF stimulation in endothelial cells. Here, we examined the interplay between these different mechanisms in VEGF-exposed endothelial cells. We documented that hsp90 binding to eNOS is, in fact, the crucial event triggering the transition from the Ca(2+)-dependent activation of eNOS to the phosphorylation-mediated potentiation of its activity by VEGF. Accordingly, we showed that early VEGF stimulation first leads to the Ca(2+)/calmodulin disruption of the caveolin-eNOS complex and promotes the association between eNOS and hsp90. eNOS-bound hsp90 can then recruit VEGF activated (phosphorylated) Akt to the complex, which in turn can phosphorylate eNOS. Further experiments in transfected COS cells expressing either wild-type or S1177A mutant eNOS led us to identify the serine 1177 as the critical residue for the hsp90-dependent Akt-mediated activation of eNOS. Finally, we documented that although the VEGF-induced phosphorylation of eNOS leads to a sustained production of NO independently of a maintained increase in [Ca(2+)](i), this late stage of eNOS activation is strictly conditional on the initial VEGF-induced Ca(2+) dependent stimulation of the enzyme. These data establish the critical temporal sequence of events leading to the sustained activation of eNOS by VEGF and suggest new ways of regulating the production of NO in response to this cytokine through the ubiquitous chaperone protein, hsp90. PMID- 11425856 TI - Expression, purification, and characterization of a soluble form of the first extracellular domain of the human type 1 corticotropin releasing factor receptor. AB - The first extracellular domain (ECD-1) of the corticotropin releasing factor (CRF) type 1 receptor, (CRFR1), is important for binding of CRF ligands. A soluble protein, mNT-CRFR1, produced by COS M6 cells transfected with a cDNA encoding amino acids 1--119 of human CRFR1 and modified to include epitope tags, binds a CRF antagonist, astressin, in a radioreceptor assay using [(125)I-d Tyr(0)]astressin. N-terminal sequencing of mNT-CRFR1 showed the absence of the first 23 amino acids of human CRFR1. This result suggests that the CRFR1 protein is processed to cleave a putative signal peptide corresponding to amino acids 1- 23. A cDNA encoding amino acids 24--119 followed by a FLAG tag, was expressed as a thioredoxin fusion protein in Escherichia coli. Following thrombin cleavage, the purified protein (bNT-CRFR1) binds astressin and the agonist urocortin with high affinity. Reduced, alkylated bNT-CRFR1 does not bind [(125)I-D Tyr(0)]astressin. Mass spectrometric analysis of photoaffinity labeled bNT-CRFR1 yielded a 1:1 complex with ligand. Analysis of the disulfide arrangement of bNT CRFR1 revealed bonds between Cys(30) and Cys(54), Cys(44) and Cys(87), and Cys(68) and Cys(102). This arrangement is similar to that of the ECD-1 of the parathyroid hormone receptor (PTHR), suggesting a conserved structural motif in the N-terminal domain of this family of receptors. PMID- 11425857 TI - The transcriptional activity of cAMP response element-binding protein-binding protein is modulated by the latency associated nuclear antigen of Kaposi's sarcoma-associated herpesvirus. AB - A multifunctional transcription co-activator, cAMP response element-binding protein-binding protein (CBP)interacts with a number of cellular factors and participates in cell growth, transformation, and development. It is also targeted by many viral proteins for their transcriptional activity or for the regulation of cellular processes. Here, we report that the C/H3 region of CBP is targeted by the latency associated nuclear antigen (LANA) of Kaposi's sarcoma-associated herpesvirus (KSHV). LANA interferes with the interaction between CBP and c-Fos, a representative C/H3 region binding, cellular transcription factor, in vivo and in vitro. In addition, we found that LANA inhibits the transcriptional activity and the in vitro histone acetyltransferase activity of CBP, suggesting that LANA modulates the global transcriptional activities of infected cells through the interaction with CBP. These results indicate that KSHV follows one of the conserved strategies, which other viruses utilize for influencing the cellular processes. PMID- 11425858 TI - Synergistic activation of the Wnt signaling pathway by Dvl and casein kinase Iepsilon. AB - Although casein kinase Iepsilon (CKIepsilon) has been shown to regulate the Wnt signaling pathway positively, its mode of action is not clear. In this study we show that CKIepsilon activates the Wnt signaling pathway in co-operation with Dvl. CKIepsilon and Axin associated with different sites of Dvl, and CKIepsilon and Dvl interacted with distinct regions on Axin. Therefore, these three proteins formed a ternary complex. Either low expression of Dvl or CKIepsilon alone did not accumulate beta-catenin, but their co-expression accumulated greatly. Dvl and CKIepsilon activated the transcriptional activity of T cell factor (Tcf) synergistically. Although the Dvl mutant that binds to Axin but not to CKIepsilon activated Tcf, it did not synergize with CKIepsilon. Another Dvl mutant that does not bind to Axin did not activate Tcf irrespective of the presence of CKIepsilon. Furthermore, Dvl and CKIepsilon co-operatively induced axis duplication of Xenopus embryos. These results indicate that Dvl and CKIepsilon synergistically activated the Wnt signaling pathway and that the binding of the complex of Dvl and CKIepsilon to Axin is necessary for their synergistic action. PMID- 11425859 TI - Regulation of receptor activator of NF-kappaB ligand-induced tartrate-resistant acid phosphatase gene expression by PU.1-interacting protein/interferon regulatory factor-4. Synergism with microphthalmia transcription factor. AB - The receptor activator of NF-kappaB ligand induces the expression of tartrate resistant acid phosphatase (TRAP) and transcription factor, PU.1-interacting protein (Pip), during osteoclastogenesis. In this paper, we have examined the role of transcription factors in the regulation of TRAP gene expression employing reporter constructs containing the promoter region of TRAP gene. Transient transfection of RAW264 cells with sequential 5'-deletions of mouse TRAP gene promoter-luciferase fusion constructs indicated that at least two sites are required for the full promoter activity. Deletion and site-directed mutation studies revealed that M-box and interferon regulatory factor element sites are critical for TPAP gene expression in the cell, suggesting that microphthalmia transcription factor (MITF) and Pip could induce the gene expression independently. Moreover, the overexpression of MITF and Pip functionally stimulated TRAP promoter activity in a synergistic manner. Analysis of the deletion mutants of Pip protein indicated that both N-terminal DNA-binding and C terminal regulatory domains are indispensable to the promoter-enhancing activity. Subcellular localization of green fluorescence protein-fused Pip and its mutant proteins indicated that the C-terminal domain is required for the translocation of Pip into the nucleus. These results suggest that Pip regulates and acts synergistically with MITF to induce the promoter activity of TRAP gene. PMID- 11425860 TI - Gas6 anti-apoptotic signaling requires NF-kappa B activation. AB - The growth arrest-specific 6 gene product Gas6 is a growth and survival factor related to protein S. Gas6 is the ligand of Axl receptor tyrosine kinase; upon binding to its receptor Gas6 activates the phosphatidylinositol 3-OH kinase (PI3K) and its downstream targets S6K and Akt. Gas6 anti-apoptotic signaling was previously shown to require functional PI3K and Akt and to involve Bad phosphorylation in serum-starved NIH 3T3 cells. Here we demonstrate that Gas6 induces a rapid and transient increase in nuclear NF-kappa B binding activity coupled to transcription activation from NF-kappa B-responsive promoters and increase in Bcl-x(L) protein level. Gas6 survival function is impaired in cells lacking p65/RelA and in NIH 3T3 cells transfected with a dominant negative I kappa B, indicating that NF-kappa B activation plays a central role in promoting survival in this system. Moreover, NF-kappa B activation can be blocked by a dominant negative Akt and by wortmannin, an inhibitor of PI3K, thus suggesting that NF-kappa B activation is a downstream event with respect to PI3K and Akt, as already described for other growth factors. In addition, we show that glycogen synthase kinase 3, which is phosphorylated in response to Gas6, can physically associate with NFKB1/p105 in living cells and can phosphorylate it in vitro. Furthermore, Gas6 treatment is coupled to a decrease in p105 protein level. Altogether these data suggest the involvement of NF-kappa B and glycogen synthase kinase 3 in Gas6 anti-apoptotic signaling and unveil a possible link between these survival pathways. PMID- 11425861 TI - Regulation of cell adhesion by polysialic acid. Effects on cadherin, immunoglobulin cell adhesion molecule, and integrin function and independence from neural cell adhesion molecule binding or signaling activity. AB - The polysialylation of neural cell adhesion molecule (NCAM) evolved in vertebrates to carry out biological functions related to changes in cell position and morphology. Many of these effects involve the attenuation of cell interactions that are not mediated through NCAM's own adhesion properties. A proposed mechanism for this global effect on cell interaction is the steric inhibition of membrane-membrane apposition based solely on polysialic acid (PSA) biophysical properties. However, it remains possible that the intrinsic binding or signaling properties of the NCAM polypeptide are also involved. To help resolve this issue, this study uses a quantitative cell detachment assay together with cells engineered to display different adhesion receptors together with a variety of polysialylated NCAM polypeptide isoforms and functional domain deletion mutations. The results obtained indicate that regulation by PSA occurs with adhesion receptors as diverse as an IgCAM, a cadherin and an integrin, and does not require NCAM functional domains other than those minimally required for polysialylation. These findings are most consistent with the cell apposition mechanism for PSA action, as this model predicts that the inhibitory effects of PSA-NCAM on cell adhesion should be independent of the nature of the adhesion system and of any intrinsic binding or signaling properties of the NCAM polypeptide itself. PMID- 11425862 TI - p75 Co-receptors regulate ligand-dependent and ligand-independent Trk receptor activation, in part by altering Trk docking subdomains. AB - Neurotrophins signal via Trk tyrosine kinase receptors and a common receptor called p75. Nerve growth factor is the cognate ligand for TrkA, brain-derived neurotrophic factor for TrkB, and neurotrophin-3 (NT-3) for TrkC. NT-3 also binds TrkA and TrkB as a heterologous ligand. All neurotrophins bind p75, which regulates ligand affinity and Trk signals. Trk extracellular domain has five subdomains: a leucine-rich motif, two cysteine-rich clusters, and immunoglobulin like subdomains IgG-C1 and IgG-C2. The IgG-C1 subdomain is surface exposed in the tertiary structure and regulates ligand-independent activation. The IgG-C2 subdomain is less exposed but regulates cognate ligand binding and Trk activation. NT-3 as a heterologous ligand of TrkA and TrkB optimally requires the IgG-C2 but also binds other subdomains of these receptors. When p75 is co expressed, major changes are observed; NGF-TrkA activation can occur also via the cysteine 1 subdomain, and brain-derived neurotrophic factor-TrkB activation requires the TrkB leucine-rich motif and cysteine 2 subdomains. We propose a two site model of Trk binding and activation, regulated conformationally by the IgG C1 subdomain. Moreover, p75 affects Trk subdomain utilization in ligand-dependent activation, possibly by conformational or allosteric control. PMID- 11425863 TI - Amelogenin-cytokeratin 14 interaction in ameloblasts during enamel formation. AB - The enamel protein amelogenin binds to the GlcNAc-mimicking peptide (GMp) (Ravindranath, R. M. H., Tam, W., Nguyen, P., and Fincham, A. G. (2000) J. Biol. Chem. 275, 39654-39661). The GMp motif is found in the N-terminal region of CK14, a differentiation marker for ameloblasts. The binding affinity of CK14 and amelogenin was confirmed by dosimetric binding of CK14 to recombinant amelogenin (rM179), and to the tyrosine-rich amelogenin polypeptide. The specific binding site for CK14 was identified in the amelogenin trityrosyl motif peptide (ATMP) of tyrosine-rich amelogenin polypeptide and specific interaction between CK14 and [(3)H]ATMP was confirmed by Scatchard analysis. Blocking rM179 with GlcNAc, GMp, or CK14 with ATMP abrogates the CK14-amelogenin interaction. CK14 failed to bind to ATMP when the third proline was substituted with threonine, as in some cases of human X-linked amelogenesis imperfecta or when tyrosyl residues were substituted with phenylalanine. Morphometry of developing teeth distinguished three phases of enamel formation; growth initiation phase (days 0-1), prolific growth phase (days 1-7), and growth cessation phase (post-day 7). Confocal microscopy revealed co-assembly of CK14/amelogenin in the perinuclear region of ameloblasts on day 0, migration of the co-assembled CK14/amelogenin to the apical region of the ameloblasts from day 1, reaching a peak on days 3-5, and a collapse of the co-assembly. Autoradiography with [(3)H]ATMP and [(3)H]GMp corroborated the dissociation of the co-assembly at the ameloblast Tomes' process. It is proposed that CK14 play a chaperon role for nascent amelogenin polypeptide during amelogenesis. PMID- 11425864 TI - A polymorphism of the human matrix gamma-carboxyglutamic acid protein promoter alters binding of an activating protein-1 complex and is associated with altered transcription and serum levels. AB - Matrix gamma-carboxyglutamic acid protein (MGP) is a mineral-binding extracellular matrix protein synthesized by vascular smooth muscle cells (VSMCs) and chondrocytes that is thought to be a key regulator of tissue calcification. In this study, we identified four polymorphisms in the promoter region of the human MGP gene. Transfection studies showed that the G-7A and T-138C polymorphisms have an important impact on in vitro promoter activity when transiently transfected into VSMCs. We found that one of these polymorphisms (T 138C) is significantly correlated with serum MGP levels in human subjects. Promoter deletion analysis showed that this polymorphism lies in a region of the promoter critical for transcription in VSMCs. This region contains a potential activating protein-1 (AP-1) binding element located between -142 and -136. We have demonstrated that the T-138C polymorphism results in altered binding of an AP-1 complex to this region. The -138T allelic variant binds AP-1 complexes consisting primarily of c-Jun, JunB and its partners Fra-1 and Fra-2 in rat VSMC. Furthermore, the -138T variant form of the promoter was induced following phorbol 12-myristate 13-acetate treatment, while the -138C variant was refractive to phorbol 12-myristate 13-acetate treatment, confirming that AP-1 factors preferentially bind to the -138T variant. This study therefore suggests that a common polymorphism of the MGP promoter influences binding of the AP-1 complex, which may lead to altered transcription and serum levels. This could have important implications for diseases such as atherosclerosis and aortic valve stenosis, since it strongly suggests a genetic basis for regulation of tissue calcification. PMID- 11425865 TI - Delineation of the clotrimazole/TRAM-34 binding site on the intermediate conductance calcium-activated potassium channel, IKCa1. AB - Selective and potent triarylmethane blockers of the intermediate conductance calcium-activated potassium channel, IKCa1, have therapeutic use in sickle cell disease and secretory diarrhea and as immunosuppressants. Clotrimazole, a membrane-permeant triarylmethane, blocked IKCa1 with equal affinity when applied externally or internally, whereas a membrane-impermeant derivative TRAM-30 blocked the channel only when applied to the cytoplasmic side, indicating an internal drug-binding site. Introduction of the S5-P-S6 region of the triarylmethane-insensitive small conductance calcium-activated potassium channel SKCa3 into IKCa1 rendered the channel resistant to triarylmethanes. Replacement of Thr(250) or Val(275) in IKCa1 with the corresponding SKCa3 residues selectively abolished triarylmethane sensitivity without affecting the affinity of the channel for tetraethylammonium, charybdotoxin, and nifedipine. Introduction of these two residues into SKCa3 rendered the channel sensitive to triarylmethanes. In a molecular model of IKCa1, Thr(250) and Val(275) line a water-filled cavity just below the selectivity filter. Structure-activity studies suggest that the side chain methyl groups of Thr(250) and Val(275) may lock the triarylmethanes in place via hydrophobic interactions with the pi-electron clouds of the phenyl rings. The heterocyclic moiety may project into the selectivity filter and obstruct the ion-conducting pathway from the inside. PMID- 11425866 TI - Kinetics of core histones in living human cells: little exchange of H3 and H4 and some rapid exchange of H2B. AB - Histones H2A and H2B form part of the same nucleosomal structure as H3 and H4. Stable HeLa cell lines expressing histones H2B, H3, and H4 tagged with green fluorescent protein (GFP) were established; the tagged molecules were assembled into nucleosomes. Although H2B-GFP was distributed like DNA, H3-GFP and H4-GFP were concentrated in euchromatin during interphase and in R-bands in mitotic chromosomes. These differences probably result from an unregulated production of tagged histones and differences in exchange. In both single cells and heterokaryons, photobleaching revealed that H2B-GFP exchanged more rapidly than H3-GFP and H4-GFP. About 3% of H2B exchanged within minutes, whereas approximately 40% did so slowly (t(1/2) approximately 130 min). The rapidly exchanging fraction disappeared in 5,6-dichloro-1-beta-d ribofuranosylbenzimidazole and so may represent H2B in transcriptionally active chromatin. The slowly exchanging fraction was probably associated with chromatin domains surrounding active units. H3-GFP and H4-GFP were assembled into chromatin when DNA was replicated, and then >80% remained bound permanently. These results reveal that the inner core of the nucleosome is very stable, whereas H2B on the surface of active nucleosomes exchanges continually. PMID- 11425867 TI - Tyrosine-phosphorylated extracellular signal--regulated kinase associates with the Golgi complex during G2/M phase of the cell cycle: evidence for regulation of Golgi structure. AB - Phosphorylation of the extracellular signal-regulated kinases (ERKs) on tyrosine and threonine residues within the TEY tripeptide motif induces ERK activation and targeting of substrates. Although it is recognized that phosphorylation of both residues is required for ERK activation, it is not known if a single phosphorylation of either residue regulates physiological functions. In light of recent evidence indicating that ERK proteins regulate substrate function in the absence of ERK enzymatic activity, we have begun to examine functional roles for partially phosphorylated forms of ERK. Using phosphorylation site--specific ERK antibodies and immunofluorescence, we demonstrate that ERK phosphorylated on the tyrosine residue (pY ERK) within the TEY activation sequence is found constitutively in the nucleus, and localizes to the Golgi complex of cells that are in late G2 or early mitosis of the cell cycle. As cells progress through metaphase and anaphase, pY ERK localization to Golgi vesicles is most evident around the mitotic spindle poles. During telophase, pY ERK associates with newly formed Golgi vesicles but is not found on there after cytokinesis and entry into G1. Increased ERK phosphorylation causes punctate distribution of several Golgi proteins, indicating disruption of the Golgi structure. This observation is reversible by overexpression of a tyrosine phosphorylation--defective ERK mutant, but not by a kinase-inactive ERK2 mutant that is tyrosine phosphorylated. These data provide the first evidence that pY ERK and not ERK kinase activity regulates Golgi structure and may be involved in mitotic Golgi fragmentation and reformation. PMID- 11425868 TI - Restricted accumulation of phosphatidylinositol 3-kinase products in a plasmalemmal subdomain during Fc gamma receptor-mediated phagocytosis. AB - Phagocytosis is a highly localized and rapid event, requiring the generation of spatially and temporally restricted signals. Because phosphatidylinositol 3 kinase (PI3K) plays an important role in the innate immune response, we studied the generation and distribution of 3' phosphoinositides (3'PIs) in macrophages during the course of phagocytosis. The presence of 3'PI was monitored noninvasively in cells transfected with chimeras of green fluorescent protein and the pleckstrin homology domain of either Akt, Btk, or Gab1. Although virtually undetectable in unstimulated cells, 3'PI rapidly accumulated at sites of phagocytosis. This accumulation was sharply restricted to the phagosomal cup, with little 3'PI detectable in the immediately adjacent areas of the plasmalemma. Measurements of fluorescence recovery after photobleaching were made to estimate the mobility of lipids in the cytosolic monolayer of the phagosomal membrane. Stimulation of phagocytic receptors induced a marked reduction of lipid mobility that likely contributes to the restricted distribution of 3'PI at the cup. 3'PI accumulation during phagocytosis was transient, terminating shortly after sealing of the phagosomal vacuole. Two factors contribute to the rapid disappearance of 3'PI: the dissociation of the type I PI3K from the phagosomal membrane and the persistent accumulation of phosphoinositide phosphatases. PMID- 11425869 TI - Role of the F-box protein Skp2 in adhesion-dependent cell cycle progression. AB - Cell adhesion to the extracellular matrix (ECM) is a requirement for proliferation that is typically lost in malignant cells. In the absence of adhesion, nontransformed cells arrest in G1 with increased levels of the cyclin dependent kinase inhibitor p27. We have reported previously that the degradation of p27 requires its phosphorylation on Thr-187 and is mediated by Skp2, an F-box protein that associates with Skp1, Cul1, and Roc1/Rbx1 to form the SCF(Skp2) ubiquitin ligase complex. Here, we show that the accumulation of Skp2 protein is dependent on both cell adhesion and growth factors but that the induction of Skp2 mRNA is exclusively dependent on cell adhesion to the ECM. Conversely, the expression of the other three subunits of the SCF(Skp2) complex is independent of cell anchorage. Phosphorylation of p27 on Thr-187 is also not affected significantly by the loss of cell adhesion, demonstrating that increased p27 stability is not dependent on p27 dephosphorylation. Significantly, ectopic expression of Skp2 in nonadherent G1 cells resulted in p27 downregulation, entry into S phase, and cell division. The ability to induce adhesion-independent cell cycle progression was potentiated by coexpressing Skp2 with cyclin D1 but not with cyclin E, indicating that Skp2 and cyclin D1 cooperate to rescue proliferation in suspension cells. Our study shows that Skp2 is a key target of ECM signaling that controls cell proliferation. PMID- 11425870 TI - Ran-binding protein 3 is a cofactor for Crm1-mediated nuclear protein export. AB - Crm1 is a member of the karyopherin family of nucleocytoplasmic transport receptors and mediates the export of proteins from the nucleus by forming a ternary complex with cargo and Ran:GTP. This complex translocates through the nuclear pores and dissociates in the cytosol. The yeast protein Yrb2p participates in this pathway and binds Crm1, but its mechanism of action has not been established. We show that the human orthologue of Yrb2p, Ran-binding protein 3 (RanBP3), acts as a cofactor for Crm1-mediated export in a permeabilized cell assay. RanBP3 binds directly to Crm1, and the complex possesses an enhanced affinity for both Ran:GTP and cargo. RanBP3 shuttles between the nucleus and the cytoplasm by a Crm1-dependent mechanism, and the Crm1--RanBP3-NES-Ran:GTP quarternary complex can associate with nucleoporins. We infer that this complex translocates through the nuclear pore to the cytoplasm where it is disassembled by RanBP1 and Ran GTPase--activating protein. PMID- 11425871 TI - The Alzheimer amyloid precursor protein (APP) and FE65, an APP-binding protein, regulate cell movement. AB - FE65 binds to the Alzheimer amyloid precursor protein (APP), but the function of this interaction has not been identified. Here, we report that APP and FE65 are involved in regulation of cell movement. APP and FE65 colocalize with actin and Mena, an Abl-associated signaling protein thought to regulate actin dynamics, in lamellipodia. APP and FE65 specifically concentrate with beta 1-integrin in dynamic adhesion sites known as focal complexes, but not in more static adhesion sites known as focal adhesions. Overexpression of APP accelerates cell migration in an MDCK cell wound--healing assay. Coexpression of APP and FE65 dramatically enhances the effect of APP on cell movement, probably by regulating the amount of APP at the cell surface. These data are consistent with a role for FE65 and APP, possibly in a Mena-containing macromolecular complex, in regulation of actin based motility. PMID- 11425872 TI - Apoptotic cleavage of cytoplasmic dynein intermediate chain and p150(Glued) stops dynein-dependent membrane motility. AB - Cytoplasmic dynein is the major minus end-directed microtubule motor in animal cells, and associates with many of its cargoes in conjunction with the dynactin complex. Interaction between cytoplasmic dynein and dynactin is mediated by the binding of cytoplasmic dynein intermediate chains (CD-IC) to the dynactin subunit, p150(Glued). We have found that both CD-IC and p150(Glued) are cleaved by caspases during apoptosis in cultured mammalian cells and in Xenopus egg extracts. Xenopus CD-IC is rapidly cleaved at a conserved aspartic acid residue adjacent to its NH(2)-terminal p150(Glued) binding domain, resulting in loss of the otherwise intact cytoplasmic dynein complex from membranes. Cleavage of CD-IC and p150(Glued) in apoptotic Xenopus egg extracts causes the cessation of cytoplasmic dynein--driven endoplasmic reticulum movement. Motility of apoptotic membranes is restored by recruitment of intact cytoplasmic dynein and dynactin from control cytosol, or from apoptotic cytosol supplemented with purified cytoplasmic dynein--dynactin, demonstrating the dynamic nature of the association of cytoplasmic dynein and dynactin with their membrane cargo. PMID- 11425873 TI - Differential dynamics of alpha 5 integrin, paxillin, and alpha-actinin during formation and disassembly of adhesions in migrating cells. AB - To investigate the mechanisms by which adhesions form and disperse in migrating cells, we expressed alpha 5 integrin, alpha-actinin, and paxillin as green fluorescent protein (GFP) fusions. All localized with their endogenous counterparts and did not perturb migration when expressed at moderate levels. alpha 5-GFP also rescued the adhesive defects in CHO B2 cells, which are alpha 5 integrin deficient. In ruffling cells, alpha 5-GFP and alpha-actinin--GFP localized prominently at the leading edge in membrane protrusions. Of the three GFP fusion proteins that we examined, paxillin was the first component to appear visibly organized in protrusive regions of the cell. When a new protrusion formed, the paxillin appeared to remodel from older to newer adhesions at the leading edge. alpha-Actinin subsequently entered adhesions, which translocated toward the cell center, and inhibited paxillin turnover. The new adhesions formed from small foci of alpha-actinin--GFP and paxillin-GFP, which grew in size. Subsequently, alpha 5 integrin entered the adhesions to form visible complexes, which served to stabilize the adhesions. alpha 5-GFP also resided in endocytic vesicles that emanated from the leading edge of protrusions. Integrin vesicles at the cell rear moved toward the cell body. As cells migrated, alpha 5 vesicles also moved from a perinuclear region to the base of the lamellipodium. The alpha 5 vesicles colocalized with transferrin receptor and FM 4-64 dye. After adhesions broke down in the rear, alpha 5-GFP was found in fibrous structures behind the cell, whereas alpha-actinin--GFP and paxillin-GFP moved up the lateral edge of retracting cells as organized structures and then dissipated. PMID- 11425874 TI - Effects of purified recombinant neural and muscle agrin on skeletal muscle fibers in vivo. AB - Aggregation of acetylcholine receptors (AChRs) in muscle fibers by nerve-derived agrin plays a key role in the formation of neuromuscular junctions. So far, the effects of agrin on muscle fibers have been studied in culture systems, transgenic animals, and in animals injected with agrin--cDNA constructs. We have applied purified recombinant chick neural and muscle agrin to rat soleus muscle in vivo and obtained the following results. Both neural and muscle agrin bind uniformly to the surface of innervated and denervated muscle fibers along their entire length. Neural agrin causes a dose-dependent appearance of AChR aggregates, which persist > or = 7 wk after a single application. Muscle agrin does not cluster AChRs and at 10 times the concentration of neural agrin does not reduce binding or AChR-aggregating activity of neural agrin. Electrical muscle activity affects the stability of agrin binding and the number, size, and spatial distribution of the neural agrin--induced AChR aggregates. Injected agrin is recovered from the muscles together with laminin and both proteins coimmunoprecipitate, indicating that agrin binds to laminin in vivo. Thus, the present approach provides a novel, simple, and efficient method for studying the effects of agrin on muscle under controlled conditions in vivo. PMID- 11425875 TI - Muscle activity and muscle agrin regulate the organization of cytoskeletal proteins and attached acetylcholine receptor (AchR) aggregates in skeletal muscle fibers. AB - In innervated skeletal muscle fibers, dystrophin and beta-dystroglycan form rib like structures (costameres) that appear as predominantly transverse stripes over Z and M lines. Here, we show that the orientation of these stripes becomes longitudinal in denervated muscles and transverse again in denervated electrically stimulated muscles. Skeletal muscle fibers express nonneural (muscle) agrin whose function is not well understood. In this work, a single application of > or = 10 nM purified recombinant muscle agrin into denervated muscles preserved the transverse orientation of costameric proteins that is typical for innervated muscles, as did a single application of > or = 1 microM neural agrin. At lower concentration, neural agrin induced acetylcholine receptor aggregates, which colocalized with longitudinally oriented beta-dystroglycan, dystrophin, utrophin, syntrophin, rapsyn, and beta 2-laminin in denervated unstimulated fibers and with the same but transversely oriented proteins in innervated or denervated stimulated fibers. The results indicate that costameres are plastic structures whose organization depends on electrical muscle activity and/or muscle agrin. PMID- 11425876 TI - Nup2p dynamically associates with the distal regions of the yeast nuclear pore complex. AB - Nucleocytoplasmic transport is mediated by the interplay between soluble transport factors and nucleoporins resident within the nuclear pore complex (NPC). Understanding this process demands knowledge of components of both the soluble and stationary phases and the interface between them. Here, we provide evidence that Nup2p, previously considered to be a typical yeast nucleoporin that binds import- and export-bound karyopherins, dynamically associates with the NPC in a Ran-facilitated manner. When bound to the NPC, Nup2p associates with regions corresponding to the nuclear basket and cytoplasmic fibrils. On the nucleoplasmic face, where the Ran--GTP levels are predicted to be high, Nup2p binds to Nup60p. Deletion of NUP60 renders Nup2p nucleoplasmic and compromises Nup2p-mediated recycling of Kap60p/Srp1p. Depletion of Ran--GTP by metabolic poisoning, disruption of the Ran cycle, or in vitro by cell lysis, results in a shift of Nup2p from the nucleoplasm to the cytoplasmic face of the NPC. This mobility of Nup2p was also detected using heterokaryons where, unlike nucleoporins, Nup2p was observed to move from one nucleus to the other. Together, our data support a model in which Nup2p movement facilitates the transition between the import and export phases of nucleocytoplasmic transport. PMID- 11425877 TI - The Dictyostelium CARMIL protein links capping protein and the Arp2/3 complex to type I myosins through their SH3 domains. AB - Fusion proteins containing the Src homology (SH)3 domains of Dictyostelium myosin IB (myoB) and IC (myoC) bind a 116-kD protein (p116), plus nine other proteins identified as the seven member Arp2/3 complex, and the alpha and beta subunits of capping protein. Immunoprecipitation reactions indicate that myoB and myoC form a complex with p116, Arp2/3, and capping protein in vivo, that the myosins bind to p116 through their SH3 domains, and that capping protein and the Arp2/3 complex in turn bind to p116. Cloning of p116 reveals a protein dominated by leucine-rich repeats and proline-rich sequences, and indicates that it is a homologue of Acan 125. Studies using p116 fusion proteins confirm the location of the myosin I SH3 domain binding site, implicate NH(2)-terminal sequences in binding capping protein, and show that a region containing a short sequence found in several G actin binding proteins, as well as an acidic stretch, can activate Arp2/3 dependent actin nucleation. p116 localizes along with the Arp2/3 complex, myoB, and myoC in dynamic actin-rich cellular extensions, including the leading edge of cells undergoing chemotactic migration, and dorsal, cup-like, macropinocytic extensions. Cells lacking p116 exhibit a striking defect in the formation of these macropinocytic structures, a concomitant reduction in the rate of fluid phase pinocytosis, a significant decrease in the efficiency of chemotactic aggregation, and a decrease in cellular F-actin content. These results identify a complex that links key players in the nucleation and termination of actin filament assembly with a ubiquitous barbed end-directed motor, indicate that the protein responsible for the formation of this complex is physiologically important, and suggest that previously reported myosin I mutant phenotypes in Dictyostelium may be due, at least in part, to defects in the assembly state of actin. We propose that p116 and Acan 125, along with homologues identified in Caenorhabditis elegans, Drosophila, mouse, and man, be named CARMIL proteins, for capping protein, Arp2/3, and myosin I linker. PMID- 11425878 TI - Cytoplasmic dynein regulation by subunit heterogeneity and its role in apical transport. AB - Despite the existence of multiple subunit isoforms for the microtubule motor cytoplasmic dynein, it has not yet been directly shown that dynein complexes with different compositions exhibit different properties. The 14-kD dynein light chain Tctex-1, but not its homologue RP3, binds directly to rhodopsin's cytoplasmic COOH-terminal tail, which encodes an apical targeting determinant in polarized epithelial Madin-Darby canine kidney (MDCK) cells. We demonstrate that Tctex-1 and RP3 compete for binding to dynein intermediate chain and that overexpressed RP3 displaces endogenous Tctex-1 from dynein complexes in MDCK cells. Furthermore, replacement of Tctex-1 by RP3 selectively disrupts the translocation of rhodopsin to the MDCK apical surface. These results directly show that cytoplasmic dynein function can be regulated by its subunit composition and that cytoplasmic dynein is essential for at least one mode of apical transport in polarized epithelia. PMID- 11425879 TI - Nucleocytoplasmic shuttling of endocytic proteins. AB - Many cellular processes rely on the ordered assembly of macromolecular structures. Here, we uncover an unexpected link between two such processes, endocytosis and transcription. Many endocytic proteins, including eps15, epsin1, the clathrin assembly lymphoid myeloid leukemia (CALM), and alpha-adaptin, accumulate in the nucleus when nuclear export is inhibited. Endocytosis and nucleocytoplasmic shuttling of endocytic proteins are apparently independent processes, since inhibition of endocytosis did not appreciably alter nuclear translocation of endocytic proteins, and blockade of nuclear export did not change the initial rate of endocytosis. In the nucleus, eps15 and CALM acted as positive modulators of transcription in a GAL4-based transactivation assay, thus raising the intriguing possibility that some endocytic proteins play a direct or indirect role in transcriptional regulation. PMID- 11425880 TI - Shaping membrane architecture: agrins in and out of the synapse. PMID- 11425881 TI - Background, but not foreground, spatial cues are taken as references for head direction responses by rat anterodorsal thalamus neurons. AB - Two populations of limbic neurons are likely neurophysiological substrates for cognitive operations required for spatial orientation and navigation: hippocampal pyramidal cells discharge selectively when the animal is in a certain place (the "firing field") in the environment, whereas head direction cells discharge when the animal orients its head in a specific, "preferred" direction. Cressant et al. (1997) showed that the firing fields of hippocampal place cells reorient relative to a group of three-dimensional objects only if these are at the periphery, but not the center of an enclosed platform. To test for corresponding responses in head direction cells, three objects were equally spaced along the periphery of a circular platform. Preferred directions were measured before and after the group of objects was rotated. (The rat was disoriented in total darkness between sessions). This was repeated in the presence or absence of a cylinder enclosing the platform. When the enclosure was present, the preferred directions of all 30 cells recorded shifted by the same angle as the objects. In the absence of the enclosure, the preferred directions did not follow the objects, remaining fixed relative to the room. These results provide a possible neurophysiological basis for observations from psychophysical experiments in humans that background, rather than foreground, cues are preferentially used for spatial orientation. PMID- 11425882 TI - Differential contributions of the basolateral and central amygdala in the acquisition and expression of conditioned relapse to cocaine-seeking behavior. AB - The amygdala is known to be a critical mediator of emotional learning in aversive and appetitive conditioning. Here we show for the first time that distinct subregions of the amygdala play unique roles in the acquisition and expression of cocaine-seeking behavior maintained by drug-paired cues in a model of relapse. Reversible inactivation of the basolateral amygdala with the sodium channel blocker tetrodotoxin disrupted both the acquisition and expression of the conditioned reinforcing effects maintained by drug-paired stimuli. However, inactivation of the central amygdala disrupted only the expression, but not the acquisition, of the conditioned reinforcing effects of drug-paired stimuli. Our results demonstrate that these nuclei participate as components of an amygdalar circuit to drive cocaine-seeking behavior produced by stimulus-reinforcement associations. PMID- 11425883 TI - Voltage-dependent sodium channels are expressed in nonspiking retinal bipolar neurons. AB - Retinal bipolar neurons transmit visual information by means of graded synaptic potentials that spread to the synaptic terminal without sodium-dependent action potentials. Although action potentials are not involved, voltage-dependent sodium channels may enhance subthreshold depolarizing potentials in the dendrites and soma of bipolar cells, as they do in other CNS neurons. We report here that voltage-dependent sodium currents are observed in a subset of bipolar neurons from goldfish retina. Single-cell reverse transcriptase-PCR identified four different sodium channel alpha subunits in goldfish bipolar cells, putatively corresponding to the mammalian voltage-gated sodium channels Na(v)1.1, Na(v)1.2, Na(v)1.3, and Na(v)1.6. The amount of sodium current was largest in cells with smaller synaptic terminals, which probably represent cone bipolar cells. Localization of sodium channel immunoreactivity in goldfish retina confirmed the expression of voltage-gated sodium channels in cone bipolar cells of both ON and OFF types. Both immunocytochemical and physiological evidence suggests that the sodium channels are localized to the soma and dendrites where they may play a role in transmission of synaptic signals, particularly in the long, thin dendrites of cone bipolar cells. PMID- 11425884 TI - Neuronal survival after CNS insult is determined by a genetically encoded autoimmune response. AB - Injury to the CNS is often followed by a spread of damage (secondary degeneration), resulting in neuronal losses that are substantially greater than might have been predicted from the severity of the primary insult. Studies in our laboratory have shown that injured CNS neurons can benefit from active or passive immunization with CNS myelin-associated antigens. The fact that autoimmune T cells can be both beneficial and destructive, taken together with the established phenomenon of genetic predisposition to autoimmune diseases, raises the question: will genetic predisposition to autoimmune diseases affect the outcome of traumatic insult to the CNS? Here we show that the survival rate of retinal ganglion cells in adult mice or rats after crush injury of the optic nerve or intravitreal injection of a toxic dosage of glutamate is up to twofold higher in strains that are resistant to the CNS autoimmune disease experimental autoimmune encephalomyelitis (EAE) than in susceptible strains. The difference was found to be attributed, at least in part, to a beneficial T-cell response that was spontaneously evoked after CNS insult in the resistant but not in the susceptible strains. In animals of EAE-resistant but not of EAE-susceptible strains devoid of mature T-cells (as a result of having undergone thymectomy at birth), the numbers of surviving neurons after optic nerve injury were significantly lower (by 60%) than in the corresponding normal animals. Moreover, the rate of retinal ganglion cell survival was higher when the optic nerve injury was preceded by an unrelated CNS (spinal cord) injury in the resistant strains but not in the susceptible strains. It thus seems that, in normal animals of EAE-resistant strains (but not of susceptible strains), the injury evokes an endogenous protective response that is T-cell dependent. These findings imply that a protective T-cell-dependent response and resistance to autoimmune disease are regulated by a common mechanism. The results of this study compel us to modify our understanding of autoimmunity and autoimmune diseases, as well as the role of autoimmunity in non autoimmune CNS disorders. They also obviously have far-reaching clinical implications in terms of prognosis and individual therapy. PMID- 11425885 TI - Kainate receptors expressed by a subpopulation of developing nociceptors rapidly switch from high to low Ca2+ permeability. AB - Dorsal root ganglion (DRG) neurons first express kainate receptor subunits, predominantly GluR5, during embryonic development. In the DRG and throughout the nervous system, substantial editing of GluR5 mRNA occurs with developmental maturation (Bernard et al., 1999). The accompanying change in Ca(2+) permeability of functional kainate receptors that is the predicted outcome of this developmental regulation of mRNA editing has not been investigated. Here we report that kainate receptors on DRG neurons from late embryonic and newborn rats are predominantly Ca(2+) permeable but then become fully Ca(2+) impermeable later in the first postnatal week. Using multiple markers for nociceptor subpopulations, we show that this switch in Ca(2+) permeability is not caused by the appearance of a new subpopulation of nociceptors with different receptor properties. Instead, the change in Ca(2+) permeability matches the time course of post-transcriptional RNA editing of GluR5 at the Q/R site within the pore of the channel, indicating that the change is probably caused by developmentally regulated RNA editing. We also report that, on the basis of the strong correlation of receptor expression with expression of the surface markers LA4, isolectin B4, and LD2, kainate receptors are present on C-fiber-type neurons projecting to lamina II of spinal cord dorsal horn. These results raise the possibility that kainate receptors in their Ca(2+)-permeable form serve a developmental role in synapse formation between this population of C-fibers and their targets in the spinal cord dorsal horn. Thereafter, the receptors may serve a new function that does not require Ca(2+) permeability. PMID- 11425886 TI - Neuroprotection from delayed postischemic administration of a metalloporphyrin catalytic antioxidant. AB - Reactive oxygen species contribute to ischemic brain injury. This study examined whether the porphyrin catalytic antioxidant manganese (III) meso-tetrakis (N ethylpyridinium-2-yl)porphyrin (MnTE-2-PyP(5+)) reduces oxidative stress and improves outcome from experimental cerebral ischemia. Rats that were subjected to 90 min focal ischemia and 7 d recovery were given MnTE-2-PyP(5+) (or vehicle) intracerebroventricularly 60 min before ischemia, or 5 or 90 min or 6 or 12 hr after reperfusion. Biomarkers of brain oxidative stress were measured at 4 hr after postischemic treatment (5 min or 6 hr). MnTE-2-PyP(5+), given 60 min before ischemia, improved neurologic scores and reduced total infarct size by 70%. MnTE 2-PyP(5+), given 5 or 90 min after reperfusion, reduced infarct size by 70-77% and had no effect on temperature. MnTE-2-PyP(5+) treatment 6 hr after ischemia reduced total infarct volume by 54% (vehicle, 131 +/- 60 mm(3); MnTE-2-PyP(5+), 300 ng, 60 +/- 68 mm(3)). Protection was observed in both cortex and caudoputamen, and neurologic scores were improved. No MnTE-2-PyP(5+) effect was observed if it was given 12 hr after ischemia. MnTE-2-PyP(5+) prevented mitochondrial aconitase inactivation and reduced 8-hydroxy-2'-deoxyguanosine formation when it was given 5 min or 6 hr after ischemia. In mice, MnTE-2-PyP(5+) reduced infarct size and improved neurologic scores when it was given intravenously 5 min after ischemia. There was no effect of 150 or 300 ng of MnTE 2-PyP(5+) pretreatment on selective neuronal necrosis resulting from 10 min forebrain ischemia and 5 d recovery in rats. Administration of a metalloporphyrin catalytic antioxidant had marked neuroprotective effects against focal ischemic insults when it was given up to 6 hr after ischemia. This was associated with decreased postischemic superoxide-mediated oxidative stress. PMID- 11425887 TI - The presynaptic function of mouse cochlear inner hair cells during development of hearing. AB - Before mice start to hear at approximately postnatal day 10, their cochlear inner hair cells (IHCs) spontaneously generate Ca(2+) action potentials. Therefore, immature IHCs could stimulate the auditory pathway, provided that they were already competent for transmitter release. Here, we combined patch-clamp capacitance measurements and fluorimetric [Ca(2+)](i) recordings to study the presynaptic function of IHCs during cochlear maturation. Ca(2+)-dependent exocytosis and subsequent endocytic membrane retrieval were already observed near the date of birth. Ca(2+) action potentials triggered exocytosis in immature IHCs, which probably activates the auditory pathway before it becomes responsive to sound. IHCs underwent profound changes in Ca(2+)-channel expression and secretion during their postnatal development. Ca(2+)-channel expression increased toward the end of the first week, providing for large secretory responses during this period and thereafter declined to reach mature levels. The efficacy whereby Ca(2+) influx triggers exocytosis increased toward maturation, such that vesicle fusion caused by a given Ca(2+) current occurred faster in mature IHCs. The observed changes in Ca(2+)-channel expression and synaptic efficacy probably reflected the ongoing synaptogenesis in IHCs that had been described previously in morphological studies. PMID- 11425888 TI - Enhanced spontaneous transmitter release is the earliest consequence of neocortical hypoxia that can explain the disruption of normal circuit function. AB - After the onset of an acute episode of arrested circulation to the brain and consequent cerebral hypoxia, EEG changes and modifications of consciousness ensue within seconds. This in part reflects the rapid effect of hypoxia on the neocortex, where oxygen deprivation leads to impaired neuronal excitability and abnormal synaptic transmission. To identify the cellular mechanisms responsible for the earliest changes in neocortical function and to determine their time course, we have used patch-in-slice recording techniques to investigate the effects of acute hypoxia on the synaptic and intrinsic properties of layer 5 neurons. Coronal slices of mouse somatosensory cortex were maintained at 37 degrees C and challenged with episodes of hypoxia (3-4 min of exposure to 95% N(2), 5% CO(2)). In recordings with cell-attached patch electrodes, activation of ATP-sensitive potassium channels first became detectable 211 +/- 11 sec (range, 185-240 sec; n = 6 patches) after the onset of hypoxia. Similar recording techniques revealed no alterations in the properties of Na(+) currents in the first 4 min after the onset of hypoxia. The earliest hypoxia-induced disturbance was a marked increase in the frequency of spontaneous EPSCs and IPSCs, which began within 15-30 sec of the removal of oxygen. This rapid synaptic effect was not sensitive to TTX and was present in Ca(2+)-free perfusate, indicating that the hypoxia had a direct influence on the vesicular release mechanisms. The incoherent, massive increase in miniature PSCs would be expected to deplete the readily releasable pool of vesicles in cortical terminals, and to thereby markedly distort the neuronal interactions that underlie normal circuit function. PMID- 11425889 TI - Differential expression of genes encoding subthreshold-operating voltage-gated K+ channels in brain. AB - The members of the three subfamilies (eag, erg, and elk) of the ether-a-go-go (EAG) family of potassium channel pore-forming subunits express currents that, like the M-current (I(M)), could have considerable influence on the subthreshold properties of neuronal membranes, and hence the control of excitability. A nonradioactive in situ hybridization (NR-ISH) study of the distribution of the transcripts encoding the eight known EAG family subunits in rat brain was performed to identify neuronal populations in which the physiological roles of EAG channels could be studied. These distributions were compared with those of the mRNAs encoding the components of the classical M-current (Kcnq2 and Kcnq3). NR-ISH was combined with immunohistochemistry to specific neuronal markers to help identify expressing neurons. The results show that each EAG subunit has a specific pattern of expression in rat brain. EAG and Kcnq transcripts are prominent in several types of excitatory neurons in the cortex and hippocampus; however, only one of these channel components (erg1) was consistently expressed in inhibitory interneurons in these areas. Some neuronal populations express more than one product of the same subfamily, suggesting that the subunits may form heteromeric channels in these neurons. Many neurons expressed multiple EAG family and Kcnq transcripts, such as CA1 pyramidal neurons, which contained Kcnq2, Kcnq3, eag1, erg1, erg3, elk2, and elk3. This indicates that the subthreshold current in many neurons may be complex, containing different components mediated by a number of channels with distinct properties and neuromodulatory responses. PMID- 11425890 TI - Pituitary adenylyl cyclase-activating peptides and alpha-amidation in olfactory neurogenesis and neuronal survival in vitro. AB - We investigated the role of amidated neuropeptides, and specifically pituitary adenylyl cyclase-activating polypeptide (PACAP), in olfactory neurogenesis and olfactory receptor neuronal survival. Using both immunohistochemistry and in situ hybridization, we find that both peptidylglycine alpha-amidating monooxygenase (PAM), the enzyme responsible for amidation and therefore activation of all amidated neuropeptides, and amidated PACAP are expressed in developing and adult olfactory epithelium. Amidated PACAP is highly expressed in proliferative basal cells and in immature olfactory neurons. The PACAP-specific receptor PAC(1) receptor is also expressed in this population, establishing that these cells can be PACAP responsive. Experiments were conducted to determine whether amidated neuropeptides, such as PACAP38, might function in olfactory neurogenesis and neuronal survival. Addition of PACAP38 to olfactory cultures increased the number of neurons to >250% of control and stimulated neuronal proliferation and survival. In primary olfactory cultures, pharmacologically decreased PAM activity, as well as neutralization of PACAP38, caused neuron-specific loss that was reversed by PACAP38. Mottled (Brindled) mice, which lack a functional ATP7A copper transporter and serve as a model for Menkes disease, provided an in vivo partial loss-of-function PAM knock-out. These mice had decreased amidated PACAP production and concomitant decreased numbers of olfactory receptor neurons. These data establish amidated peptides and specifically PACAP as having important roles in proliferation in the olfactory system and suggest that a similar function exists in vivo. PMID- 11425891 TI - The OMP-lacZ transgene mimics the unusual expression pattern of OR-Z6, a new odorant receptor gene on mouse chromosome 6: implication for locus-dependent gene expression. AB - Reporter gene expression in the olfactory epithelium of H-lacZ6 transgenic mice mimics the cell-selective expression pattern known for some odorant receptor genes. The transgene construct in these mice consists of the lacZ coding region, driven by the proximal olfactory marker protein (OMP) gene promoter, and shows expression in a zonally confined subpopulation of olfactory neurons. To address mechanisms underlying the odorant receptor-like expression pattern of the lacZ construct, we analyzed the transgene-flanking region and identified OR-Z6, the first cloned odorant receptor gene that maps to mouse chromosome 6. OR-Z6 bears the highest sequence similarity (85%) to a human odorant receptor gene at the syntenic location on human chromosome 7. We analyzed the expression pattern of OR Z6 in olfactory tissues of H-lacZ6 mice and show that it bears strong similarities to that mapped for beta-galactosidase. Expression of both genes in olfactory neurons is primarily restricted to the same medial subregion of the olfactory epithelium. Axons from both neuronal subpopulations project to the same ventromedial aspect of the anterior olfactory bulbs. Furthermore, colocalization analyses in H-lacZ6 mice demonstrate that OR-Z6-reactive glomeruli receive axonal input from lacZ-positive neurons as well. These results suggest that the expression of both genes is coordinated and that transgene expression in H-lacZ6 mice is regulated by locus-dependent mechanisms. PMID- 11425892 TI - MIP-1alpha, MCP-1, GM-CSF, and TNF-alpha control the immune cell response that mediates rapid phagocytosis of myelin from the adult mouse spinal cord. AB - The slow immune response in the adult mammalian CNS results in slow myelin phagocytosis along degenerating white matter after injury. This has important consequences for axon regeneration because of the presence of axon growth inhibitors in myelin. In addition, abnormal immune cell responses in the CNS lead to demyelinating disease. Lysophosphatidylcholine (LPC) can induce an inflammatory response in the CNS, producing rapid demyelination without much damage to adjacent cells. In this study, we searched for the molecular switches that turn on this immune cell response. Using reverse transcription PCR analysis, we show that mRNA expression of macrophage inflammatory protein-1alpha (MIP 1alpha), macrophage chemotactic protein-1 (MCP-1), granulocyte macrophage-colony stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-alpha) in the spinal cord is rapidly and transiently upregulated after intraspinal injection of LPC. Neutralizing these signaling molecules with function-blocking antibodies suppresses recruitment of T-cells, neutrophils, and monocytes into the spinal cord, as well as significantly reduces the number of phagocytic macrophages and the demyelination induced by LPC. These findings will have important implications for CNS regeneration and demyelinating disease. PMID- 11425893 TI - Taxol induces apoptosis in cortical neurons by a mechanism independent of Bcl-2 phosphorylation. AB - Bcl-2, an antiapoptotic protein, protects cells against many but not all forms of apoptosis. For example, Bcl-2 does not protect non-neuronal cells against taxol, a microtubule-stabilizing agent. The underlying mechanism for the ineffectiveness of Bcl-2 against taxol has been the subject of intense interest. Data from non neuronal cells indicate that taxol-induced apoptosis requires activation of N terminal c-Jun protein kinase (JNK) that phosphorylates and inactivates Bcl-2. This suggests the interesting possibility that the apoptotic activity of JNK may be caused by phosphorylation of Bcl-2 and inhibition of the antiapoptotic activity of Bcl-2. Here we report that taxol induces apoptosis in cortical neurons but by a mechanism significantly different from that in non-neuronal cells. In contrast to human embryonic kidney 293 cells, expression of wild-type Bcl-2 in cortical neurons protected against taxol-induced apoptosis, and taxol did not induce Bcl-2 phosphorylation. Furthermore, cortical neurons express high basal JNK activity, and taxol did not stimulate total JNK activity. However, taxol activated a subpool of JNK in the nucleus and stimulated c-Jun phosphorylation. JNK inhibition or expression of a dominant-negative c-Jun abrogated taxol-induced apoptosis in cortical neurons, suggesting a role for JNK and JNK-mediated transcription in taxol-stimulated apoptosis. Furthermore, taxol induced apoptosis in cortical neurons required inhibition of phosphatidylinositol 3-kinase signaling. These data suggest that taxol induces apoptosis in neurons by a mechanism quite distinct from that of non-neuronal cell lines and emphasize the importance of elucidating apoptotic mechanisms specific for neurons in the CNS. PMID- 11425894 TI - Activation of the nuclear factor-kappaB is a key event in brain tolerance. AB - The transcription factor nuclear factor-kappaB (NFkappaB) is an ubiquitously expressed inducible regulator of a broad range of genes and plays a pivotal role in cell death and survival pathways. Three models of brain tolerance (ischemic, epileptic, and polyunsaturated fatty acid-induced preconditioning), known to confer resistance to neurons against ischemia or status epilepticus, were used to determine whether NFkappaB mediated the late preconditioning. A sublethal 3 min ischemia, a dose of 5 mg/kg kainic acid (KA5) or 500 nmol of linolenic acid (LIN500) led to a rapid increase of NFkappaB DNA-binding activity and nuclear translocation of p65 and p50 subunits of NFkappaB in neurons. Pretreatment with the NFkappaB inhibitor diethyldithiocarbamate or kappaB decoy DNA blocked the increased DNA-binding activity and the nuclear translocation of NFkappaB and abolished the neuroprotective effects of different delayed preconditionings against severe ischemia or epilepsy. The inhibition of NFkappaB observed in rats preconditioned with 3 min ischemia, KA5 or LIN500 treatments compared with ischemic or epileptic controls was correlated with the prevention of the inducible degradation of the inhibitory protein IkappaBalpha. Preconditioning probably inhibits the activation of NFkappaB by interfering with a pathway that leads to the direct transcriptional activation of IkappaBalpha by NFkappaB itself. The present work provides evidence that activation of NFkappaB is a crucial step in the signal transduction pathway that underlies the development of brain tolerance and may open new strategies in the prevention of cerebral diseases, such as ischemia or epilepsy. PMID- 11425895 TI - Caspase-activated DNase/DNA fragmentation factor 40 mediates apoptotic DNA fragmentation in transient cerebral ischemia and in neuronal cultures. AB - Nuclear changes, including internucleosomal DNA fragmentation, are characteristic features of neuronal apoptosis resulting from transient cerebral ischemia and related brain insults for which the molecular mechanism has not been elucidated. Recent studies suggest that a caspase-3-mediated mechanism may be involved in the process of nuclear degradation in ischemic neurons. In this study, we cloned from rat brain a homolog cDNA encoding caspase-activated deoxyribonuclease (CAD)/DNA fragmentation factor 40 (DFF40), a 40 kDa nuclear enzyme that is activated by caspase-3 and promotes apoptotic DNA degradation. Subsequently, we investigated the role of CAD/DFF40 in the induction of internucleosomal DNA fragmentation in the hippocampus in a rat model of transient global ischemia and in primary neuronal cultures under ischemia-like conditions. At 8-72 hr after ischemia, CAD/DFF40 mRNA and protein were induced in the degenerating hippocampal CA1 neurons. CAD/DFF40 formed a heterodimeric complex in the nucleus with its natural inhibitor CAD (ICAD) and was activated after ischemia in a delayed manner (>24 hr) by caspase-3, which translocated into the nucleus and cleaved ICAD. Furthermore, an induced CAD/DFF40 activity was detected in nuclear extracts in both in vivo and in vitro models, and the DNA degradation activity of CAD/DFF40 was inhibited by purified ICAD protein. These results strongly suggest that CAD/DFF40 is the endogenous endonuclease that mediates caspase-3-dependent internucleosomal DNA degradation and related nuclear alterations in ischemic neurons. PMID- 11425896 TI - Age-related impairment of synaptic transmission but normal long-term potentiation in transgenic mice that overexpress the human APP695SWE mutant form of amyloid precursor protein. AB - We have studied synaptic function in a transgenic mouse strain relevant to Alzheimer's disease (AD), overexpressing the 695 amino acid isoform of human amyloid precursor protein with K670N and M671L mutations (APP(695)SWE mice), which is associated with early-onset familial AD. Aged-transgenic mice had substantially elevated levels of Abeta (up to 22 micromol/gm) and displayed characteristic Abeta plaques. Hippocampal slices from 12-month-old APP(695)SWE transgenic animals displayed reduced levels of synaptic transmission in the CA1 region when compared with wild-type littermate controls. Inclusion of the ionotropic glutamate receptor antagonist kynurenate during preparation of brain slices abolished this deficit. At 18 months of age, a selective deficit in basal synaptic transmission was observed in the CA1 region despite treatment with kynurenate. Paired-pulse facilitation and long-term potentiation (LTP) were normal in APP(695)SWE transgenic mice at both 12 and 18 months of age. Thus, although aged APP(695)SWE transgenic mice have greatly elevated levels of Abeta protein, increased numbers of plaques, and reduced basal synaptic transmission, LTP can still be induced and expressed normally. We conclude that increased susceptibility to excitotoxicity rather than a specific effect on LTP is the primary cause of cognitive deficits in APP(695)SWE mice. PMID- 11425897 TI - Boundary formation and compartition in the avian diencephalon. AB - The diencephalon comprises three functionally distinct regions: synencephalon, dorsal thalamus, and ventral thalamus. Patterning of the diencephalon has been proposed to involve subdivision of its anteroposterior axis into segments, neuromeres or prosomeres (Bergquist and Kallen, 1954; Vaage, 1969; Figdor and Stern, 1993; Rubenstein et al., 1994; Redies et al., 2000; Yoon et al., 2000). However, the number and sequence of diencephalic neuromeres, or even their existence, are uncertain. We have examined the proposed subdivisions by morphology, gene expression, acquisition of boundary-specific phenotypes, and cell lineage restriction. We find that at stage 16 in chick the diencephalon is divided into synencephalon and parencephalon. The synencephalon exhibits neuromeric morphology, expresses Prox, and acquires neuromere boundary properties at its interface with both the midbrain and the parencephalon. Although the mesencephalic/synencephalic boundary restricts cell mixing, the synencephalic/parencephalic boundary does not. Similarly, there is no lineage restriction between the parencephalon and the more rostral forebrain (secondary prosencephalon). Subdivision of the parencephalon into ventral and dorsal thalamus involves the formation of a narrow intraparencephalic territory, the zona limitans intrathalamica (zli). This is correlated with the acquisition of cell lineage restriction at both anterior and posterior borders of the zli, the appearance of boundary-specific properties, and Gbx2 and Dlx2 expression in dorsal thalamic and ventral thalamic territories, respectively. At stage 22, the synencephalon is divided into two domains, distinguished by differential gene expression and tissue morphology, but associated with neither a boundary phenotype nor cell lineage restriction. Our results suggest that the diencephalon does not have an overt segmental pattern. PMID- 11425898 TI - Hes1 and Hes5 activities are required for the normal development of the hair cells in the mammalian inner ear. AB - The mammalian inner ear contains two sensory organs, the cochlea and vestibule. Their sensory neuroepithelia are characterized by a mosaic of hair cells and supporting cells. Cochlear hair cells differentiate in four rows: a single row of inner hair cells (IHCs) and three rows of outer hair cells (OHCs). Recent studies have shown that Math1, a mammalian homolog of Drosophila atonal is a positive regulator of hair cell differentiation. The basic helix-loop-helix (bHLH) genes Hes1 and Hes5 (mammalian hairy and Enhancer-of-split homologs) can influence cell fate determination by acting as negative regulators to inhibit the action of bHLH positive regulators. We show by using reverse transcription-PCR analysis that Hes1, Hes5, and Math1 are expressed in the developing mouse cochleae. In situ hybridization revealed a widespread expression of Hes1 in the greater epithelial ridge (GER) and in lesser epithelial ridge (LER) regions. Hes5 is predominantly expressed in the LER, in supporting cells, and in a narrow band of cells within the GER. Examination of cochleae from Hes1(-/-) mice showed a significant increase in the number of IHCs, whereas cochleae from Hes5(-/-) mice showed a significant increase in the number of OHCs. In the vestibular system, targeted deletion of Hes1 and to a lesser extent Hes5 lead to formation of supernumerary hair cells in the saccule and utricle. The supernumerary hair cells in the mutant mice showed an upregulation of Math1. These data indicate that Hes1 and Hes5 participate together for the control of inner ear hair cell production, likely through the negative regulation of Math1. PMID- 11425899 TI - NMDA receptor and nitric oxide synthase activation regulate polysialylated neural cell adhesion molecule expression in adult brainstem synapses. AB - Here we report that synapses in the adult dorsal vagal complex, a gateway for many primary afferent fibers, express a high level of the polysialylated neural cell adhesion molecule (PSA-NCAM). We show that electrical stimulation of the vagal afferents causes a rapid decrease of PSA-NCAM expression both in vivo and in acute slices. Inhibition of NMDA receptor activity completely prevented the decrease. Blockade of calmodulin activation, neuronal nitric oxide (NO) synthase, or soluble guanylyl cyclase and chelation of extracellular NO mimicked this inhibition. Our data provide a mechanistic framework for understanding how activity-linked stimulation of the NMDA-NO-cGMP pathway induces rapid changes in PSA-NCAM expression, which may be associated with long-term depression. PMID- 11425900 TI - Neuronal cyclic AMP controls the developmental loss in ability of axons to regenerate. AB - Unlike neonatal axons, mammalian adult axons do not regenerate after injury. Likewise, myelin, a major factor in preventing regeneration in the adult, inhibits regeneration from older but not younger neurons. Identification of the molecular events responsible for this developmental loss of regenerative capacity is believed key to devising strategies to encourage regeneration in adults after injury. Here, we report that the endogenous levels of the cyclic nucleotide, cAMP, are dramatically higher in young neurons in which axonal growth is promoted both by myelin in general and by a specific myelin component, myelin-associated glycoprotein (MAG), than in the same types of neurons that, when older, are inhibited by myelin-MAG. Inhibiting a downstream effector of cAMP [protein kinase A (PKA)] prevents myelin-MAG promotion from young neurons, and elevating cAMP blocks myelin-MAG inhibition of neurite outgrowth in older neurons. Importantly, developmental plasticity of spinal tract axons in neonatal rat pups in vivo is dramatically reduced by inhibition of PKA. Thus, the switch from promotion to inhibition by myelin-MAG, which marks the developmental loss of regenerative capacity, is mediated by a developmentally regulated decrease in endogenous neuronal cAMP levels. PMID- 11425901 TI - Neuregulin signaling regulates neural precursor growth and the generation of oligodendrocytes in vitro. AB - Neuregulin 1 (Nrg-1) isoforms have been shown to influence the emergence and growth of oligodendrocytes, the CNS myelin-forming cells. We have investigated how Nrg-1 signaling of ErbB receptors specifically controls the early stages of oligodendrocyte generation from multipotential neural precursors (NPs). We show here that embryonic striatal NPs express multiple Nrg-1 transcripts and proteins as well as their specific receptors, ErbB2 and ErbB4, but not ErbB3. The major isoform synthesized by striatal NPs is a transmembrane type III isoform called cysteine-rich domain Nrg-1. To examine the biological effect of Nrg-1, we added soluble ErbB3 (sErbB3) to growing neurospheres. This inhibitor of Nrg-1 bioactivity decreased mitosis of NPs and increased their apoptosis, resulting in a significant reduction in neurosphere size and number. When NPs were induced to migrate and differentiate by adhesion of neurospheres to the substratum, the level of type III isoforms detected by RT-PCR and Western blot decreased in parallel with a reduction in Nrg-1 fluorescence intensity in differentiating astrocytes, neurons, and oligodendrocytes. Pretreatment of growing neurospheres with sErbB3 induced a threefold increase in the proportion of oligodendrocytes generated from NPs migrating out of the neurosphere. This effect was not observed with an unrelated soluble receptor. Addition of sErbB3 during NP growth and differentiation enhanced oligodendrocyte maturation as shown by expression of galactocerebroside and myelin basic protein. We propose that both type III Nrg-1 signaling and soluble ErbB receptors modulate oligodendrocyte development from NPs. PMID- 11425902 TI - Programmed cell death of developing mammalian neurons after genetic deletion of caspases. AB - An analysis of programmed cell death of several populations of developing postmitotic neurons after genetic deletion of two key members of the caspase family of pro-apoptotic proteases, caspase-3 and caspase-9, indicates that normal neuronal loss occurs. Although the amount of cell death is not altered, the death process may be delayed, and the cells appear to use a nonapoptotic pathway of degeneration. The neuronal populations examined include spinal interneurons and motor, sensory, and autonomic neurons. When examined at both the light and electron microscopic levels, the caspase-deficient neurons exhibit a nonapoptotic morphology in which nuclear changes such as chromatin condensation are absent or reduced; in addition, this morphology is characterized by extensive cytoplasmic vacuolization that is rarely observed in degenerating control neurons. There is also reduced terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling in dying caspase-deficient neurons. Despite the altered morphology and apparent temporal delay in cell death, the number of neurons that are ultimately lost is indistinguishable from that seen in control animals. In contrast to the striking perturbations in the morphology of the forebrain of caspase-deficient embryos, the spinal cord and brainstem appear normal. These results are consistent with the growing idea that the involvement of specific caspases and the occurrence of caspase-independent programmed cell death may be dependent on brain region, cell type, age, and species or may be the result of specific perturbations or pathology. PMID- 11425903 TI - Progressive cerebellar, auditory, and esophageal dysfunction caused by targeted disruption of the frizzled-4 gene. AB - Wnt signaling has been implicated in the control of cell proliferation and in synapse formation during neural development, and these actions are presumed to be mediated by frizzled receptors. In this paper we report the phenotype of mice carrying a targeted deletion of the frizzled-4 (fz4) gene. fz4(-/-) mice exhibit three distinct defects: (1) progressive cerebellar degeneration associated with severe ataxia, (2) absence of a skeletal muscle sheath around the lower esophagus associated with progressive esophageal distension and dysfunction, and (3) progressive deafness caused by a defect in the peripheral auditory system unaccompanied by loss of hair cells or other auditory neurons. As assayed using a lacZ knock-in reporter, fz4 is widely expressed within the CNS. In particular, fz4 is expressed in cerebellar Purkinje cells, esophageal skeletal muscle, and cochlear inner hair cells, and the absence of Fz4 in these cells is presumed to account for the fz4(-/-) phenotype. In contrast to the early cell proliferation and patterning effects classically ascribed to Wnts, the auditory and cerebellar phenotypes of fz4(-/-) mice implicate Frizzled signaling in maintaining the viability and integrity of the nervous system in later life. PMID- 11425904 TI - Intact aggrecan and fragments generated by both aggrecanse and metalloproteinase like activities are present in the developing and adult rat spinal cord and their relative abundance is altered by injury. AB - Aggrecan is a large proteoglycan (PG) that has been grouped with different PG families on the basis of its physical characteristics. These families include the chondroitin sulfate PGs, which appear to inhibit the migration of cells and axons during development. Although aggrecan has been studied primarily in cartilage, in the present study, tissue samples from developing, mature, and injured-adult rat spinal cords were used to determine whether aggrecan is present in the mammalian spinal cord. By the use of Western blot analysis, tissues were probed with aggrecan-specific antibodies (ATEGQV, TYKHRL, and LEC-7) and aggrecan-specific neoepitope antibodies (NITEGE, FVDIPEN, and TFKEEE) to identify full-length aggrecan and several fragments. Unlike many other aggrecan gene family members, aggrecan species were similar in embryonic day 14, postnatal day 1, and adult spinal cords. Spinal cord injury caused significant decreases in aggrecan. Partial recovery in some aggrecan species was evident by 2 weeks after injury. The presence of specific aggrecan neoepitopes suggested that aggrecan is cleaved in the spinal cord by both a disintegrin and metalloproteinase thrombospondin (also known as aggrecanase) and metalloproteinase-like activities. Many aggrecan species found in the spinal cord were similar to species in cartilage. Additional antibodies were used to identify two other aggrecan gene family members, neurocan and brevican, in the adult spinal cord. These studies present novel information on the aggrecan core protein species and enzymes involved in aggrecan cleavage in vivo in the rat spinal cord throughout development and after injury. They also provide the basis for investigating the function of aggrecan in the spinal cord. PMID- 11425905 TI - Adult neuronal regeneration induced by transgenic integrin expression. AB - In a variety of adult CNS injury models, embryonic neurons exhibit superior regenerative performance when compared with adult neurons. It is unknown how young neurons extend axons in the injured adult brain, in which adult neurons fail to regenerate. This study shows that cultured adult neurons do not adapt to conditions that are characteristic of the injured adult CNS: low levels of growth promoting molecules and the presence of inhibitory proteoglycans. In contrast, young neurons readily adapt to these same conditions, and adaptation is accompanied by an increase in the expression of receptors for growth-promoting molecules (receptors of the integrin family). Surprisingly, the regenerative performance of adult neurons can be restored to that of young neurons by gene transfer-mediated expression of a single alpha-integrin. PMID- 11425906 TI - Neuronal hypertrophy in the neocortex of patients with temporal lobe epilepsy. AB - The underlying cause of neocortical involvement in temporal lobe epilepsy (TLE) remains a fundamental and unanswered question. Magnetic resonance imaging has shown a significant loss in temporal lobe volume, and it has been proposed that neocortical circuits are disturbed functionally because neurons are lost. The present study used design-based stereology to estimate the volume and cell number of Brodmann's area 38, a region commonly resected in anterior temporal lobectomy. Studies were conducted on the neocortex of patients with or without hippocampal sclerosis (HS). Results provide the surprising finding that TLE patients have significant atrophy of neocortical gray matter but no loss of neurons. Neurons are also significantly larger, dendritic trees appear sparser, and spine density is noticeably reduced in TLE specimens compared with controls. The increase in neuronal density we found in TLE patients is therefore attributable to large neurons occupying a much smaller volume than in normal brain. Neurons in the underlying white matter are also increased in size but, in contrast to other reports, are not significantly elevated in number or density. Neuronal hypertrophy affects HS and non-HS brains similarly. The reduction in neuropil and its associated elements therefore appears to be a primary feature of TLE, which is not secondary to cell loss. In both gray and white matter, neuronal hypertrophy means more perikaryal surface area is exposed for synaptic contacts and emerges as a hallmark of this disease. PMID- 11425907 TI - A code for behavioral inhibition on the basis of color, but not motion, in ventrolateral prefrontal cortex of macaque monkey. AB - To examine the neural mechanism for behavioral inhibition, we recorded single cell activity in macaque ventrolateral prefrontal cortex, which is known to receive visual information directly from the inferotemporal cortex. In response to a moving random pattern of colored dots, monkeys had to make a go or no-go response. In the color condition, green indicated go, whereas red indicated no go, regardless of the motion direction; in the motion condition, upward indicated go, whereas downward indicated no-go, regardless of the color. Approximately one half of the visual cells were go/no-go differential. A majority of these cells (64/73) showed differential activity only in the color condition; they responded nondifferentially in the motion condition, although the same set of stimuli was used. We classified these cells as "go type" (n = 41) and "no-go type" (n = 23) depending on the color for which they showed a stronger response. Interestingly, in both types of cells, the differential effects were observed only for the no-go indicating color. Compared with the nondifferential responses in the motion condition, go-type cells in the color condition showed weaker responses to the no go-indicating color, whereas their responses to the go-indicating color were similar; in contrast, no-go type cells showed stronger responses to the no-go indicating color, whereas their responses to the go-indicating color were similar. Both types of cells did not show any activity change during the actual execution of the go or no-go response. These results suggest that neurons in ventrolateral prefrontal cortex contribute to stimulus-response association in complex task situations by inhibiting behavioral responses on the basis of visual information from the ventral stream. PMID- 11425908 TI - Perceptually bistable three-dimensional figures evoke high choice probabilities in cortical area MT. AB - The role of the primate middle temporal area (MT) in depth perception was examined by considering the trial-to-trial correlations between neuronal activity and reported depth sensations. A set of moving random dots portrayed a cylinder rotating about its principal axis. In this structure-from-motion stimulus, the direction of rotation is ambiguous and the resulting percept undergoes spontaneous fluctuations. The stimulus can be rendered unambiguous by the addition of binocular disparities. We trained monkeys to report the direction of rotation in a set of these stimuli, one of which had zero disparity. Many disparity-selective neurons in area MT are selective for the direction of rotation defined by disparity. Across repeated presentations of the ambiguous (zero-disparity) stimulus, there was a correlation between neuronal firing and the reported direction of rotation, as found by Bradley et al. (1998). Quantification of this effect using choice probabilities (Britten et al., 1996) allowed us to demonstrate that the correlation cannot be explained by eye movements, behavioral biases, or attention to spatial location. MT neurons therefore appear to be involved in the perceptual decision process. The mean choice probability (0.67) was substantially larger than that reported for MT neurons in a direction discrimination task (Britten et al., 1996). This implies that MT neurons make a different contribution to the two tasks. For the depth task, either the pool of neurons used is smaller or the correlation between neurons in the pool is larger. PMID- 11425909 TI - Psychological stress increases hippocampal mineralocorticoid receptor levels: involvement of corticotropin-releasing hormone. AB - We investigated whether acute stressors regulate functional properties of the hippocampal mineralocorticoid receptor (MR), which acts inhibitory on hypothalamic-pituitary-adrenocortical activity. Exposure of rats to forced swimming or novelty evoked a significant rise in density of MR immunoreactivity in all hippocampal subfields after 24 hr, whereas exposure to a cold environment was ineffective. Time course analysis revealed that the effect of forced swimming on MR peaked at 24 hr and returned to control levels between 24 and 48 hr. In pyramidal neurons of CA2 and CA3, marked rises were already observed after 8 hr. Radioligand binding assays showed that corticotropin-releasing hormone (CRH) injected intracerebroventricularly into adrenalectomized rats also produced a rise in hippocampal MR levels; an effect for which the presence of corticosterone, but not dexamethasone, at the time of injection was a prerequisite. Moreover, pretreatment with the CRH receptor antagonist (d Phe(12),Nle(21,38),alpha-Me-Leu(37))-CRH(12-41) blocked the effect of forced swimming on hippocampal MR levels. To investigate whether the rise in MR levels had any functional consequences for HPA regulation, 24 hr after forced swimming, a challenge test with the MR antagonist RU 28318 was conducted. The forced swimming exposed rats showed an enhanced MR-mediated inhibition of HPA activity. This study identifies CRH as an important regulator of MR, a pathway with marked consequence for HPA axis regulation. We conclude that the interaction between CRH and MR presents a novel mechanism involved in the adaptation of the brain to psychologically stressful events. PMID- 11425910 TI - Reversible inactivation of the dorsal nucleus of the lateral lemniscus reveals its role in the processing of multiple sound sources in the inferior colliculus of bats. AB - Neurons in the inferior colliculus (IC) that are excited by one ear and inhibited by the other [excitatory-inhibitory (EI) neurons] can code interaural intensity disparities (IIDs), the cues animals use to localize high frequencies. Although EI properties are first formed in a lower nucleus and imposed on some IC cells via an excitatory projection, many other EI neurons are formed de novo in the IC. By reversibly inactivating the dorsal nucleus of the lateral lemniscus (DNLL) in Mexican free-tailed bats with kynurenic acid, we show that the EI properties of many IC cells are formed de novo via an inhibitory projection from the DNLL on the opposite side. We also show that signals excitatory to the IC evoke an inhibition in the opposite DNLL that persists for tens of milliseconds after the signal has ended. During that period, strongly suppressed EI cells in the IC are deprived of inhibition from the DNLL and respond to binaural signals as weakly inhibited or monaural cells. By relieving inhibition at the IC, we show that an initial binaural signal essentially reconfigures the circuit and thereby allows IC cells to respond to trailing binaural signals that were inhibitory when presented alone. Thus, DNLL innervation creates a property in the IC that is not possessed by lower neurons or by collicular EI neurons that are not innervated by the DNLL. That property is a change in responsiveness to binaural signals, a change dependent on the reception of an earlier sound. These features suggest that the circuitry linking the DNLL with the opposite central nucleus of the IC is important for the processing of IIDs that change over time, such as the IIDs generated by moving stimuli or by multiple sound sources that emanate from different regions of space. PMID- 11425911 TI - Tuning to interaural time differences across frequency. AB - Interaural time differences (ITDs) are an important cue for azimuthal sound localization. Sensitivity to this cue depends on temporal synchrony to the waveform (i.e., phase locking) that begins in the hair cells and is relayed to the neural comparators. The synchrony function is low-pass. Therefore, it is expected that neural tuning to ITDs will become narrower with frequency according to a 1/frequency function. To test this, we measured ITD tuning across frequency in neurons from the superior olivary complex, the dorsal nucleus of the lateral lemniscus, the inferior colliculus, the auditory thalamus, and the auditory cortex. For some neurons in each nucleus, the ITD tuning width did become systematically narrower by the expected 1/frequency relationship. However, in other neurons the ITD tuning width was nearly constant across frequency. Constant ITD tuning width was infrequently observed in neurons of the superior olivary complex but was common in neurons in structures above the superior olivary complex. The nearly constant ITD tuning was caused both by sharper ITD tuning at low frequencies and broader tuning at higher frequencies within the low-frequency band. Neurons with nearly constant tuning to ITDs may be the mechanism underlying the perception of ITDs in humans in which just-noticeable differences to changes in ITD decrease by less than the 1/frequency prediction. PMID- 11425912 TI - Synaptic currents generating the inhibitory surround of ganglion cells in the mammalian retina. AB - The receptive field (RF) of retinal ganglion cells (RGCs) consists of an excitatory central region, the RF center, and an inhibitory peripheral region, the RF surround. It is still unknown in detail which inhibitory interneurons (horizontal or amacrine cells) and which inhibitory circuits (presynaptic or postsynaptic) generate the RF surround. To study surround inhibition, light evoked whole-cell currents were recorded from RGCs of the isolated, intact rabbit retina. The RFs were stimulated with light or dark spots of increasing diameters and with annular light stimuli. Direct inhibitory currents could be isolated by voltage clamping ganglion cells close to the Na(+)/K(+) reversal potential. They mostly represent an input from GABAergic amacrine cells that contribute to the inhibitory surround of ganglion cells. This direct inhibitory input and its physiological function were also investigated by recording light-evoked action potentials of RGCs in the current-clamp mode and by changing the intracellular Cl(-) concentration. The excitatory input of the ganglion cells could be isolated by voltage clamping ganglion cells at the Cl(-) reversal potential. Large light spots and annular light stimuli caused a strong attenuation of the excitatory input. Both GABA(A) receptors and GABA(C) receptors contributed to this inhibition, and picrotoxinin was able to completely block it. Together, these results show that the RF surround of retinal ganglion cells is mediated by a combination of direct inhibitory synapses and presynaptic surround inhibition. PMID- 11425913 TI - Contrasting effects of ibotenate lesions of the paraventricular nucleus and subparaventricular zone on sleep-wake cycle and temperature regulation. AB - The suprachiasmatic nucleus (SCN), the circadian pacemaker for the brain, provides a massive projection to the subparaventricular zone (SPZ), but the role of the SPZ in circadian processes has received little attention. We examined the effects on circadian rhythms of sleep, body temperature, and activity in rats of restricted ibotenic acid lesions of the ventral or dorsal SPZ that spared the immediately adjacent paraventricular hypothalamic nucleus (PVH) and the SCN. Ventral SPZ lesions caused profound reduction of measures of circadian index of sleep (by 90%) and locomotor activity (75% reduction) but had less effect on body temperature (50% reduction); dorsal SPZ lesions caused greater reduction of circadian index of body temperature (by 70%) but had less effect on circadian index of locomotor activity (45% reduction) or sleep (<5% reduction). The loss of circadian regulation of body temperature or sleep was replaced by a strong ultradian rhythm (period approximately 3 hr). Lesions of the PVH, immediately dorsal to the SPZ, had no significant effect on any circadian rhythms that we measured, nor did the lesions affect the baseline body temperature. However, the fever response after intravenous injection of lipopolysaccharide (5 microg/kg) was markedly decreased in the rats with PVH lesions (66.6%) but not dorsal SPZ lesions. These results indicate that circadian rhythms of sleep and body temperatures are regulated by separate neuronal populations in the SPZ, and different aspects of thermoregulation (circadian rhythm and fever response) are controlled by distinct anatomical substrates. PMID- 11425914 TI - The alpha(2a)-adrenergic receptor plays a protective role in mouse behavioral models of depression and anxiety. AB - The noradrenergic system is involved in the regulation of many physiological and psychological processes, including the modulation of mood. The alpha(2) adrenergic receptors (alpha(2)-ARs) modulate norepinephrine release, as well as the release of serotonin and other neurotransmitters, and are therefore potential targets for antidepressant and anxiolytic drug development. The current studies were undertaken to examine the role of the alpha(2A) subtype of alpha(2)-AR in mouse behavioral models of depression and anxiety. We have observed that the genetic knock-out of the alpha(2A)-AR makes mice less active in a modified version of Porsolt's forced swim test and insensitive to the antidepressant effects of the tricyclic drug imipramine in this paradigm. Furthermore, alpha(2A) AR knock-out mice appear more anxious than wild-type C57 Bl/6 mice in the rearing and light-dark models of anxiety after injection stress. These findings suggest that the alpha(2A)-AR may play a protective role in some forms of depression and anxiety and that the antidepressant effects of imipramine may be mediated by the alpha(2A)-AR. PMID- 11425915 TI - Dissociation between light-induced phase shift of the circadian rhythm and clock gene expression in mice lacking the pituitary adenylate cyclase activating polypeptide type 1 receptor. AB - The circadian clock located in the suprachiasmatic nucleus (SCN) organizes autonomic and behavioral rhythms into a near 24 hr time that is adjusted daily to the solar cycle via a direct projection from the retina, the retinohypothalamic tract (RHT). This neuronal pathway costores the neurotransmitters PACAP and glutamate, which seem to be important for light-induced resetting of the clock. At the molecular level the clock genes mPer1 and mPer2 are believed to be target for the light signaling to the clock. In this study, we investigated the possible role of PACAP-type 1 receptor signaling in light-induced resetting of the behavioral rhythm and light-induced clock gene expression in the SCN. Light stimulation at early night resulted in larger phase delays in PACAP-type 1 receptor-deficient mice (PAC1(-)/-) compared with wild-type mice accompanied by a marked reduction in light-induced mPer1, mPer2, and c-fos gene expression. Light stimulation at late night induced mPer1 and c-fos gene expression in the SCN to the same levels in both wild type and PAC1(-)/- mice. However, in contrast to the phase advance seen in wild-type mice, PAC1(-)/- mice responded with phase delays after photic stimulation. These data indicate that PAC1 receptor signaling participates in the gating control of photic sensitivity of the clock and suggest that mPer1, mPer2, and c-fos are of less importance for light-induced phase shifts at night. PMID- 11425916 TI - Brain-derived neurotrophic factor increases in the uninjured dorsal root ganglion neurons in selective spinal nerve ligation model. AB - Nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are two major members of the neurotrophin family. Using immunohistochemistry and in situ hybridization histochemistry, we examined the effect of L5 spinal nerve ligation (SPNL), a neuropathic pain model, on the expression of BDNF in the uninjured L4 dorsal root ganglion (DRG). After L5 SPNL, both immunoreactivity for BDNF and the hybridization intensity for BDNF mRNA increased mainly in the small- and medium sized neurons. The percentage of BDNF mRNA-expressing neurons increased in the ipsilateral L4 DRG compared with the contralateral DRG from the third to 28th day after ligation. A significantly greater number of BDNF-immunoreactive neurons were observed in the ipsilateral L4 DRG than contralateral side 14 d after ligation. To test the contribution of BDNF to the thermal hyperalgesia produced in this model, we intrathecally injected anti-BDNF antibody at third day after ligation. This treatment clearly attenuated thermal hyperalgesia for a few hours. Almost all BDNF mRNA-expressing neurons coexpressed trkA, a high-affinity NGF receptor, mRNA. The percentage of BDNF mRNA-expressing cells of trkA cells significantly increased in the ipsilateral L4 DRG 14 d after ligation. Furthermore, we examined the contribution of NGF on this phenotypic change using ELISA, Northern blot analysis, and anti-NGF antibody. NGF content in the ipsilateral L4 DRG linearly increased and reached a statistical significant level 14 d after L5 SPNL. Moreover, at this time point, the increase in NGF mRNA was observed in the ipsilateral L5 DRG and sciatic nerve, but not in the ipsilateral L4 DRG or L4 spinal nerve. Local application of anti-NGF antibody to the L4 spinal nerve beside the L5 spinal nerve-ligation site prevented the development of thermal hyperalgesia for 5 d after ligation. Our data suggest that BDNF, which increased in the uninjured L4 DRG neurons, acts as a sensory neuromodulator in the dorsal horn and contributes to thermal hyperalgesia in this neuropathic pain model. The contribution of locally synthesized NGF to thermal hyperalgesia was also demonstrated. These dynamic alterations in the expression and content of BDNF and NGF in the uninjured DRG neurons might be involved in the pathomechanisms of neuropathic pain. PMID- 11425917 TI - Expression of striatal preprotachykinin mRNA in symptomatic and asymptomatic 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine-exposed monkeys is related to parkinsonian motor signs. AB - Striatal preprotachykinin (PPT) gene expression and [(3)H]mazindol binding were examined in monkeys exposed to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Some animals (n = 5) became moderately to severely parkinsonian after receiving large doses of MPTP over 9-30 d and remained symptomatic for a relatively short time (3 weeks to 3 months; acutely symptomatic group). A second group of animals (n = 5) received low doses of MPTP (1.5-12 months), developed cognitive impairments but displayed no gross motor deficits (asymptomatic group), and were killed 3-12 months after their final dose of MPTP. Other animals became moderately to severely parkinsonian after receiving escalating doses of MPTP (>6 months; n = 4) or high doses of MPTP (<1 month; n = 1) and remained symptomatic for 2.5-5.75 years (chronically symptomatic group). All MPTP-treated animals had extensive losses of [(3)H]mazindol binding in dorsal striatal sensorimotor regions with asymptomatic animals generally having a lesser degree of damage. However, PPT mRNA levels differed sharply among treatment groups. Symptomatic animals (acutely and chronically parkinsonian) had significantly decreased PPT mRNA levels in most striatal regions. In asymptomatic animals, PPT mRNA expression was not significantly different from that measured in control animals, despite decreases in [(3)H]mazindol binding in some striatal regions of similar magnitude to those observed in symptomatic animals. These observations suggest that PPT gene expression may be directly related to expression of parkinsonian motor symptomatology regardless of duration of MPTP exposure, duration of the parkinsonism, or extent of dopamine denervation. These results imply that the direct striatal output circuit may have a greater contribution to expression of parkinsonian symptomatology than proposed previously. PMID- 11425918 TI - Distinct changes in cortical acetylcholine and noradrenaline efflux during contingent and noncontingent performance of a visual attentional task. AB - Optimization of cognitive processing may depend on specific and distinct functions of the cortical cholinergic and noradrenergic systems. This investigation dissociates functions of cortical acetylcholine (ACh) and noradrenaline (NA) in arousal and visual attention by simultaneously measuring ACh and NA efflux in the rat prefrontal cortex during sustained attentional performance. The five-choice serial reaction time task was used to provide a continuous assessment of visuospatial attention. Previous studies using this task have established a critical role for the cortical cholinergic system in the detection of visual targets. However, selective lesions of the locus coeruleus noradrenergic system impair performance only when additional attentional demands are placed on the subject by distractors or temporally unpredictable targets. To test the hypothesis that the cortical noradrenergic system is particularly sensitive to novel task contingencies, we also assessed NA and ACh efflux in rats that been trained previously on the task but for whom the instrumental contingency coupling responding with stimulus detection and reward was abolished. Cortical ACh efflux showed a robust and task-related increase during established contingent performance. This response was significantly attenuated in noncontingent subjects, although it still exceeded pretask values. In contrast, NA efflux only increased transiently in contingent subjects after task onset but showed sustained elevations in noncontingent subjects on the first day when contingencies were changed. These data also implicate cortical ACh in aspects of attentional functioning but highlight a specific involvement of the cortical noradrenergic system in detecting shifts in the predictive relationship between instrumental action and reinforcement. PMID- 11425919 TI - Glutamatergic afferents from the hippocampus to the nucleus accumbens regulate activity of ventral tegmental area dopamine neurons. AB - Several studies have shown that the mesolimbic dopamine (DA) system is strongly influenced by the ventral subiculum (vSub) of the hippocampus. To examine whether this occurs by activation of DA neuron firing, the effects of chemical stimulation of the vSub on ventral tegmental area (VTA) DA neuron activity were examined using extracellular single-unit recordings. Infusions of NMDA into the vSub increased the number of spontaneously firing DA cells recorded per electrode track, while having no effect on firing rate or burst firing. This response was abolished by intranucleus accumbens (NAc) infusions of the glutamate receptor antagonist kynurenic acid. This effect did not involve the prefrontal cortex, because infusions of tetrodotoxin into the prefrontal cortex did not affect the increase in spontaneously active DA cells. Infusions of either kynurenic acid into the NAc or tetrodotoxin into the vSub decreased the firing rate and burst firing of DA neurons without altering the number of spontaneously active DA neurons. These data show that glutamatergic afferents from the vSub to the NAc exert a potent excitatory effect on VTA DA neurons, influencing both DA neuron population activity and the regulation of the firing properties of these neurons. As a result, dysfunctions in hippocampal circuitries may contribute to the hyperexcitable state of the DA system that is present in schizophrenia. PMID- 11425920 TI - Energy hypometabolism in posterior cingulate cortex of Alzheimer's patients: superficial laminar cytochrome oxidase associated with disease duration. AB - Among brain regions affected in Alzheimer's disease (AD), the posterior cingulate shows the earliest and largest decrement in energy metabolism. Positron emission tomography (PET) studies have shown that these decrements appear before the onset of memory deficits or other symptoms in persons at genetic risk for AD. This study compares in vivo imaging results and in situ postmortem analyses by examining the posterior cingulate (area 23) in 15 AD patients and 13 age-matched nondemented controls using quantitative cytochrome oxidase histochemistry as an intracellular measure of oxidative energy metabolic capacity. Each of the six layers of the posterior cingulate demonstrated a decline in cytochrome oxidase activity in AD relative to controls, whereas adjacent motor cortex showed no significant differences. This decrement did not appear to be mainly secondary to nonspecific decrement in mitochondrial enzymes, oxidative stress, cell loss, or histopathology. The cytochrome oxidase decrement was most severe in the superficial layer I (-39%), which demonstrated a correlation to disease duration. Covariance analyses suggest that superficial laminas undergo a functional uncoupling from the deeper layers of posterior cingulate cortex in AD, whereas no such effects are found in motor cortex or controls. These findings expand on previous results from PET studies by illuminating the layer-specific cytochrome oxidase contributions to energy hypometabolism. The findings suggest a decrement of cytochrome oxidase in posterior cingulate cortex, with progressive reduction within the superficial laminas linked to disease duration. Such decrement could contribute to some of the behavioral symptoms displayed by AD patients. This decrement appeared greater in women. PMID- 11425921 TI - Distribution of gustatory sensitivities in rat taste cells: whole-cell responses to apical chemical stimulation. AB - Several taste transduction mechanisms have been demonstrated in mammals, but little is known about their distribution within and across receptor cells. We recorded whole-cell responses of 120 taste cells of the rat fungiform papillae and soft palate maintained within the intact epithelium in a modified Ussing chamber, which allowed us to flow tastants across the apical membrane while monitoring the activity of the cell with a patch pipette. Taste stimuli were: 0.1 m sucrose, KCl, and NH(4)Cl, 0.032 m NaCl, and 3.2 mm HCl and quinine hydrochloride (QHCl). When cells were held at their resting potentials, taste stimulation resulted in conductance changes; reversible currents >5 pA were considered reliable responses. Sucrose and QHCl produced a decrease in outward current and membrane conductance, whereas NaCl, KCl, NH(4)Cl, and HCl elicited inward currents accompanied by increased conductance. Combinations of responses to pairs of the four basic stimuli (sucrose, NaCl, HCl, and QHCl) across the 71 84 cells tested with each pair were predictable from the probabilities of responses to individual stimuli, indicating an independent distribution of sensitivities. Of 62 cells tested with all four basic stimuli, 59 responded to at least one of the stimuli; 16 of these (27.1%) responded to only one, 20 (33.9%) to two, 15 (25.4%) to three, and 8 (13.6%) to all of the basic stimuli. Cells with both inward (Na(+)) and outward (K(+)) voltage-activated currents were significantly more broadly tuned to gustatory stimuli than those with only inward currents. PMID- 11425923 TI - Solving the mysteries of narcolepsy: the hypocretin story. PMID- 11425924 TI - Polyglutamine aggregates in SCA6 Purkinje cells: a tail of two toxicities. PMID- 11425925 TI - The cost of delaying treatment in multiple sclerosis: what is lost is not regained. PMID- 11425926 TI - PRISMS-4: Long-term efficacy of interferon-beta-1a in relapsing MS. AB - BACKGROUND: The PRISMS study demonstrated significant clinical and MRI benefit at 2 years for interferon-beta-1a, 22 and 44 mcg thrice weekly (tiw), compared with placebo in relapsing-remitting MS. Years 3 and 4 extension study results are reported. METHODS: Patients initially receiving placebo were randomized to blinded interferon-beta-1a, 22 or 44 mcg tiw (n = 172; crossover group); others continued blinded treatment with their originally assigned dose, 22 mcg (Rx22 group) or 44 mcg (Rx44 group) tiw (n = 167 per group). Patients had 3- to 6-month clinical and annual MRI assessments. RESULTS: Relapse rates for 4 years were 1.02 (crossover), 0.80 (Rx22, p < 0.001), and 0.72 (Rx44, p < 0.001); the dose effect approached significance (p = 0.069; risk ratio, 0.88; 95% CI, 0.76-1.01). Crossover groups showed reductions in relapse count, MRI activity, and lesion burden accumulation with interferon-beta-1a compared with their placebo period (p < 0.001 both doses). Time to sustained disability progression was prolonged by 18 months in the Rx44 group compared with the crossover group (p = 0.047). Rx22 and Rx44 reduced new T2 lesion number and lesion burden compared with crossover (p < 0.001); Rx44 was superior to Rx22 on several clinical and MRI outcomes. Persistent neutralizing antibodies developed in 14.3% (Rx44) and 23.7% (Rx22) of patients and were associated with reduced efficacy. CONCLUSIONS: Clinical and MRI benefit continued for both doses up to 4 years, with evidence of dose response. Outcomes were consistently better for patients treated for 4 years than for patients in crossover groups. Efficacy decreased with neutralizing antibody formation. PMID- 11425927 TI - Handicap after acute whiplash injury: a 1-year prospective study of risk factors. AB - BACKGROUND: Exposure to a whiplash injury implies a risk for development of chronic disability and handicap, with reported frequencies ranging from 0% to 50% in follow-up studies. The exact risk for development of chronic whiplash syndrome is not known. OBJECTIVE: To prospectively determine the sensitivity and specificity of five possible predictors for handicap following a whiplash injury. METHODS: In a 1-year prospective study of persons with acute whiplash injury (n = 141) and control subjects who had acute ankle distortion (n = 40), pain intensity, number of nonpainful neurologic complaints, cervical mobility, workload during extension and flexion of the neck, and results of psychometric assessment were recorded. The consecutively sampled injured persons were assessed with structured and semistructured questionnaires, and underwent neurologic examination after 1 week and 1, 3, 6, and 12 months. After 3 to 4 years, participants with whiplash injury were questioned about legal issues. RESULTS: After 1 year, 11 (7.8%) persons with whiplash injury had not returned to usual level of activity or work. The best single estimator of handicap was the cervical range-of-motion test, which had a sensitivity of 73% and a specificity of 91% (p < 0.01, Cox regression analysis). Accuracy and specificity increased to 94% and 99% when combined with pain intensity and other complaints. This increase was gained at the expense of a reduced sensitivity. Initiation of lawsuit within first month after injury did not influence recovery. CONCLUSION: The cervical range-of-motion test has a high sensitivity in prediction of handicap after acute whiplash injury. The value of cervical range-of-motion test is further improved by additional recording of symptoms and pain intensity. PMID- 11425928 TI - Temporal lobe epilepsy due to hippocampal sclerosis in pediatric candidates for epilepsy surgery. AB - OBJECTIVE: To characterize the clinical, EEG, MRI, and histopathologic features and explore seizure outcome in pediatric candidates for epilepsy surgery who have temporal lobe epilepsy (TLE) caused by hippocampal sclerosis (HS). METHODS: The authors studied 17 children (4 to 12 years of age) and 17 adolescents (13 to 20 years of age) who had anteromesial temporal resection between 1990 and 1998. RESULTS: All patients had seizures characterized by decreased awareness and responsiveness. Automatisms were typically mild to moderate in children and moderate to marked in adolescents. Among adolescents, interictal spikes were almost exclusively unilateral anterior temporal, as opposed to children in whom anterior temporal spikes were associated with mid/posterior temporal, bilateral temporal, extratemporal, or generalized spikes in 60% of cases. MRI showed hippocampal sclerosis on the side of EEG seizure onset in all patients. Fifty four percent of children and 56% of adolescents had significant asymmetry of total hippocampal volumes, whereas the remaining patients had only focal atrophy of the hippocampal head or body. Subtle MRI abnormalities of ipsilateral temporal neocortex were seen in all children and 60% of adolescents studied with FLAIR images. On histopathology, there was an unexpectedly high frequency of dual pathology with mild to moderate cortical dysplasia as well as HS, seen in 79% of children and adolescents. Seventy-eight percent of patients were free of seizures at follow-up (mean, 2.6 years). A tendency for lower seizure-free outcome was observed in patients with bilateral temporal interictal sharp waves or bilateral HS on MRI. The presence of dual pathology did not portend poor postsurgical outcome. CONCLUSIONS: TLE caused by HS similar to those in adults were seen in children as young as 4 years of age. Focal hippocampal atrophy seen on MRI often was not reflected in total hippocampal volumetry. Children may have an especially high frequency of dual pathology, with mild to moderate cortical dysplasia as well as HS, and MRI usually, but not always, predicts this finding. Postsurgical seizure outcome is similar to that in adult series. PMID- 11425929 TI - Relationship of flumazenil and glucose PET abnormalities to neocortical epilepsy surgery outcome. AB - BACKGROUND: Cortical areas showing abnormal glucose metabolism and [(11)C]flumazenil (FMZ) binding are commonly seen on PET scans of patients with intractable partial epilepsy, but it is unclear whether these must be totally resected to achieve seizure control. OBJECTIVE: To analyze whether the extent of cortex showing 2-deoxy-2-[(18)F]fluoro-D-glucose (FDG) or FMZ PET abnormalities correlates with the outcome of resective epilepsy surgery. METHODS: Cortical FDG and FMZ PET abnormalities in 15 young patients (mean age, 12.2 +/- 7.0 years) with intractable partial epilepsy of neocortical origin were marked as regions with abnormal asymmetry using an objective semiautomated software package. These marked regions were then projected and measured on the brain surface reconstructed from the coregistered high-resolution MRI. Following cortical resection, the size of nonresected cortex with preoperative PET abnormalities was also measured (calculated separately for marked areas in the lobe of seizure onset as defined by long-term video EEG monitoring, and in remote cortical areas). Extent of preoperative PET abnormalities and postoperative nonresected cortex abnormalities on PET were correlated with outcome scores. RESULTS: Large preoperative FMZ PET abnormalities were associated with poor outcome (r = 0.57; p = 0.025). Larger areas of nonresected cortex with preoperative FMZ PET abnormalities in the lobe of seizure onset were also associated with worse outcome in the whole group (r = 0.66; p = 0.007) as well as in patients with extratemporal resection (r = 0.73; p = 0.007), and in those with no lesion on MRI (r = 0.60; p = 0.049). Patients with seizure-free outcome had significantly smaller nonresected cortex with preoperative FMZ PET abnormalities than those who continued to have seizures (p = 0.022). No significant correlations between nonresected FDG PET abnormalities and surgical outcome were found. CONCLUSIONS: Extensive cortical abnormalities on FMZ PET predict poor outcome in neocortical epilepsy surgery. Resection of FMZ abnormalities in the lobe of seizure onset is associated with excellent outcome even in the absence of a structural lesion. In contrast, although FDG PET abnormalities regionalized the epileptogenic area, their size was not related to the extent of epileptogenic tissue to be removed. PMID- 11425930 TI - Quantitative magnetic resonance characterization of mesial temporal sclerosis in childhood. AB - OBJECTIVE: To investigate whether quantitative MR techniques can be used to distinguish between mesial temporal sclerosis in patients with a history of prolonged febrile convulsion and in patients without such a history. METHODS: Quantitative hippocampal T2 relaxometry, hippocampal volumetry, and single voxel (1)H-MRS data were acquired from 16 children who subsequently underwent temporal lobe resections for intractable temporal lobe epilepsy and histologically were shown to have sclerosis of the horn of Ammon. Eight children had a history of prolonged febrile convulsion in early childhood and eight children had other or no associations. RESULTS: Patients with a history of prolonged febrile convulsion had smaller hippocampi (p = 0.02) and prolonged T2 relaxation time (p = 0.03) ipsilateral to the seizure focus when compared with patients without such a history. There was also more side-to-side asymmetry of T2 relaxation time (p = 0.004) and hippocampal volume (p = 0.02) in the patients with a history of prolonged febrile convulsion than in those with other or no associations. No differences between the groups were identified using (1)H-MRS. CONCLUSIONS: These data support the view that there are at least two types of mesial temporal sclerosis. There may be several pathogenetic pathways from initial insult to later mesial temporal sclerosis, and these pathways are, at least in part, dependent on the initial insult. PMID- 11425931 TI - Treatment of stroke in rat with intracarotid administration of marrow stromal cells. AB - OBJECTIVE: To measure the therapeutic efficacy for the treatment of stroke with intra-arterial administration of bone marrow stromal cells (MSC). BACKGROUND: MSC have characteristics of stem and progenitor cells. The hypothesis that MSC injected into the internal carotid artery after stroke enter into ischemic brain and improve neurologic recovery was tested. METHODS: Twenty-five adult Wistar rats were subjected to transient (2-hour) middle cerebral artery occlusion alone (n = 9), or treated with intracarotid arterial injection of 200 microL phosphate buffered saline (n = 8) or 2 x 10(6) MSC in 200 microL phosphate-buffered saline (n = 8) 1 day after ischemia. MSC were harvested and isolated from additional adult rats and then cultured and labeled with bromodeoxyuridine. Rats were subjected to neurologic functional tests (adhesive-removal, modified neurologic severity scores) before and at 1, 7, and 14 days after middle cerebral artery occlusion. Immunohistochemistry was used to identify cell-specific proteins of bromodeoxyuridine-reactive MSC. RESULTS: Bromodeoxyuridine-reactive cells ( approximately 21% of 2 x 10(6) injected MSC) distributed throughout the territory of the middle cerebral artery by 14 days after ischemia. Some bromodeoxyuridine reactive cells expressed proteins characteristic of astrocytes and neurons. Rats with intra-arterial transplantation of MSC exhibited improvement on the adhesive removal test (p < 0.05) and the modified neurologic severity scores (p < 0.05) at 14 days compared with controls. CONCLUSIONS: MSC injected intra-arterially are localized and directed to the territory of the middle cerebral artery, and these cells foster functional improvement after cerebral ischemia. PMID- 11425932 TI - Aneurysmal SAH: cognitive outcome and structural damage after clipping or coiling. AB - BACKGROUND: Aneurysmal subarachnoid hemorrhage (SAH) and surgical clipping of intracranial aneurysms are associated with substantial morbidity and mortality. OBJECTIVE: To compare cognitive outcome and structural damage in patients with aneurysmal SAH treated with surgical clipping or endovascular coiling. METHODS: Forty case-matched pairs of patients with aneurysmal SAH treated by surgical clipping or endovascular coiling were prospectively assessed by use of a battery of cognitive tests. Twenty-three case-matched pairs underwent MRI 1 year after the procedure. Matching was based on grade of SAH on admission, location of aneurysm, age, and premorbid IQ. RESULTS: Both groups were impaired in all cognitive domains when compared with age-matched healthy control subjects. Comparison of cognitive outcome between the two groups indicated an overall trend toward a poorer cognitive outcome in the surgical group, which achieved significance in four tests. MRI showed focal encephalomalacia exclusively in the surgical group. This group also had a significantly higher incidence of single or multiple small infarcts within the vascular territory of the aneurysm, but both groups had similar incidence of large infarcts and global ischemic damage. CONCLUSION: Endovascular treatment may cause less structural brain damage than surgery and have a more favorable cognitive outcome. However, cognitive outcome appears to be dictated primarily by the complications of SAH. PMID- 11425933 TI - Predominant involvement of a particular group of fingers due to small, cortical infarction. AB - BACKGROUND: Small cortical strokes may cause predominant weakness restricted to a particular group of fingers. However, clinical, radiologic, and etiopathogenetical studies have rarely been done in these patients. METHOD: The author analyzed clinical and radiologic features of 12 patients with small cortical infarcts who had predominant weakness of particular fingers. RESULTS: The patients were divided into two groups: Eight patients had predominant involvement of ulnar-sided fingers (PIUF), and four had predominant involvement of radial-sided fingers (PIRF). Sensory symptoms were also present in eight patients, and the severity correlated with that of motor impairment. Radiologic clinical correlation showed that the lesions related to PIUF were located significantly more medially than those associated with PIRF in the presumed hand representation area of the motor cortex. In addition, the PIUF was closely associated with severe proximal vessel stenosis or occlusion, whereas the PIRF was often related to emboligenic stroke. CONCLUSIONS: The results agree with traditionally alleged topography for fingers in the human motor cortex: ulnar fingers-medial and radial fingers-lateral. However, whether this result indicates a point-to-point somatotopical representation or differential threshold of excitability in individual neurons remains unknown. Sensory topography seems to be closely associated with that of motor function in individual subjects. The motor cortex representing ulnar-sided fingers may be a borderzone area between large arteries. PMID- 11425934 TI - Midlife vascular risk factors and late-life mild cognitive impairment: A population-based study. AB - OBJECTIVE: To evaluate the impact of midlife elevated serum cholesterol levels and blood pressure on the subsequent development of mild cognitive impairment (MCI) and to investigate the prevalence of MCI in elderly Finnish population, applying the MCI criteria devised by the Mayo Clinic Alzheimer's Disease Research Center. BACKGROUND: MCI has been considered as a predictor of AD. Vascular risk factors may be important in the development of cognitive impairment and AD. However, the role of vascular risk factors in MCI and the prevalence of MCI still remain virtually unknown. METHODS: Subjects were derived from random, population based samples previously studied in surveys carried out in 1972, 1977, 1982, and 1987. After an average follow-up of 21 years, 1,449 subjects aged 65 to 79 years were reexamined in 1998. RESULTS: Eighty-two subjects, 6.1% of the population (average age, 72 years) met the criteria for MCI. Midlife elevated serum cholesterol level (> or =6.5 mmol/L) was a significant risk factor for MCI (OR, 1.9; 95% CI, 1.2 to 3.0, adjusted for age and body mass index); the effect of systolic blood pressure approached significance. CONCLUSION: Data point to a role for midlife vascular risk factors in the development of MCI in late life. PMID- 11425935 TI - Prevalence of Alzheimer's disease in very elderly people: a prospective neuropathological study. AB - BACKGROUND: No previous autopsy-controlled, prospective, and population-based studies are available on the prevalence of AD in very elderly people. OBJECTIVE: To study the point prevalence of neuropathologically defined AD in a population of people at least 85 years of age, stratified according to their APOE genotype. METHODS: A population-based sample of 532 (of a total population of 601) elderly Finnish individuals, aged 85 years or more, were clinically tested for dementia in 1991 (with follow-up studies of the survivors in 1994, 1996, and 1999) and genotyped for APOE. An autopsy involving neuropathologic diagnosis of AD according to modified consensus criteria was performed in 118 of 198 deceased subjects who had been demented on April 1, 1991, and in 62 of 201 nondemented individuals. RESULTS: The prevalence of neuropathologically defined AD was 33%, whereas the prevalence of clinically diagnosed AD was 16%. There was a highly significant (p < 0.001) association between the APOE epsilon4 allele and AD: Sixty-three percent of APOE epsilon4 carriers and 20% of noncarriers had neuropathologic AD. The respective figures in subjects aged 90 years or more were 71 and 22%. CONCLUSIONS: The prevalence of neuropathologically defined AD is higher than that reported in most previous studies based on clinical diagnosis. The discrepancy between the neuropathologic and clinical diagnoses of AD in very elderly subjects may affect the results of population-based studies. The APOE genotype has a strong effect on the prevalence of neuropathologically defined AD, even after 90 years of age. PMID- 11425936 TI - Gender and age modify the association between APOE and AD-related neuropathology. AB - OBJECTIVE: To assess the impact of apolipoprotein E (APOE) polymorphism on AD related neurofibrillary tangle (NFT) formation and senile plaques (SP). METHODS: A sample of 729 routine autopsy brains (359 men, 370 women; age range, 60 to 99 years) was investigated. All brains were classified neuropathologically according to a procedure permitting differentiation of six NFT stages and three SP stages. APOE genotyping was performed on all cases. RESULTS: The epsilon4 allele of APOE was associated not only with SP (p < 0.0001) but also with NFT formation (p < 0.0001). The effect of the epsilon4 allele on NFT formation was noted at ages > or =80 years (p < 0.0001) but not between ages 60 and 79 years (p = 0.12). An association between the epsilon4 allele and SP for women was found at ages 60 to 79 years (p < 0.0001) but not at > or =80 years of age (p = 0.063). By comparison, men showed an association in both age categories (p = 0.001 and p = 0.001). CONCLUSION: The results confirm the association between the epsilon4 allele and both types of AD-related lesions and show that this association is differentially modified by age and gender. PMID- 11425937 TI - Corticobasal degeneration and progressive supranuclear palsy share a common tau haplotype. AB - OBJECTIVE: To analyze the association of polymorphisms in the tau gene with pathologically confirmed corticobasal degeneration (CBD). BACKGROUND: The authors previously described an extended tau haplotype (H1) that covers the human tau gene and is associated with the development of progressive supranuclear palsy (PSP). The authors now extend this analysis to CBD, a neurodegenerative condition with clinical and neuropathologic similarities to PSP. Like PSP, CBD is associated with accumulation of aggregates containing the 4-repeat isoforms of tau. Because of difficulty in diagnosis of CBD, the authors only analyzed cases with pathologically confirmed CBD. METHODS: The authors collected 57 unrelated, neuropathologically confirmed cases of CBD. Tau sequencing in these cases failed to show the presence of pathogenic mutations. Polymorphisms that spanned the tau gene were analyzed in all CBD cases and controls. RESULTS: Analyzing tau polymorphisms in CBD cases showed that the frequency of H1 and H1/H1 was significantly increased when analyzing all cases and when separating by country of origin. H1 frequency in all CBD cases was 0.921, compared with a control frequency of 0.766 (X(2) = 9.1, p = 0.00255 [1df], OR 3.56 [8.43 > CI 95% > 1.53]). The H1/H1 frequency was also significantly higher at 0.842 compared with 0.596 in age-matched controls (X(2) = 17.42, p = 0.00016, 2df), OR 3.61 [7.05 > CI 95% > 1.85]). CONCLUSIONS: The CBD tau association described here suggests that PSP and CBD share a similar cause, although the pathogenic mechanism behind the two diseases leads to a different clinical and pathologic phenotype. PMID- 11425938 TI - Use and cost of outpatient visits of AD patients: CERAD XXII. AB - OBJECTIVE: To determine the probability, frequency, and cost of outpatient visits of patients with AD in the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) as a function of stage of dementia and institutional status. METHODS: Clinical information on 388 patients with AD enrolled in CERAD who had no serious comorbidities at baseline and for whom the stage of disease and institutional status were known, were linked to Health Care Financing Administration Physician/Supplier and Outpatient Standard Analytic (institutional outpatient) files for 1991 through 1995. None was registered in a health maintenance organization. Repeated measures regression models were used to examine the relationship of stage of disease to probability, frequency, and cost of outpatient visits for institutionalized and noninstitutionalized patients, with demographic characteristics and calendar time controlled. RESULTS: The annual proportion of patients with AD and a Medicare-reimbursed outpatient visit ranged from 81% to 95% and was not related to stage of dementia or institutional status. Among those with at least one outpatient visit, however, those living at home had fewer visits than did those in institutions, but their number of visits increased as dementia worsened. Those in institutions had a larger number of outpatient visits, but these did not vary significantly by stage of dementia. Neither location of residence nor stage affected the cost of outpatient visits. CONCLUSION: Among those with an outpatient visit, the frequency of visits and their relationship to stage of disease depends on institutional status. PMID- 11425939 TI - Freezing of gait in PD: prospective assessment in the DATATOP cohort. AB - OBJECTIVE: To study the development of freezing of gait in PD. BACKGROUND: Freezing of gait is a common, disabling, and poorly understood symptom in PD. METHODS: The authors analyzed data from 800 patients with early PD from the Deprenyl and Tocopherol Antioxidative Therapy of Parkinsonism (DATATOP) clinical trial who were assigned either placebo, deprenyl, tocopherol, or the combination of deprenyl and tocopherol. The primary outcome measure was the time from randomization until the freezing of gait score on the Unified Parkinson's Disease Rating Scale (UPDRS) became positive. RESULTS: Fifty-seven patients (7.1%) had freezing of gait at study entry and 193 (26%) of the remaining patients experienced the symptom by the end of the follow-up period. Those with freezing of gait at baseline had significantly more advanced disease than those without the symptom, as measured by total UPDRS and Hoehn and Yahr stage. High baseline risk factors for developing freezing of gait during the follow-up period were the onset of PD with a gait disorder; higher scores of rigidity, postural instability, bradykinesia and speech; and longer disease duration. In contrast, tremor was strongly associated with a decreased risk for freezing of gait. At the end of follow-up, the signs most strongly associated with the freezing phenomenon were gait, balance, and speech disorders, not rigidity or bradykinesia. Deprenyl treatment was strongly associated with a decreased risk for developing freezing of gait; tocopherol had no effect. CONCLUSIONS: Freezing of gait is directly related to duration of PD. Risk factors at onset of disease are the absence of tremor and PD beginning as a gait disorder. The development of freezing of gait in the course of the illness is strongly associated with the development of balance and speech problems, less so with the worsening of bradykinesia, and is not associated with the progression of rigidity. These results support the concept that the freezing phenomenon is distinct from bradykinesia. Deprenyl, in the absence of L-dopa, was found to be an effective prophylactic treatment and should be considered for patients with PD who have an onset of gait difficulty. PMID- 11425940 TI - Small-fiber dysfunction in trigeminal neuralgia: carbamazepine effect on laser evoked potentials. AB - BACKGROUND: In patients with trigeminal neuralgia, results of clinical examination of sensory function are normal. Reflex and evoked potential studies have already provided information on large-afferent (non-nociceptive) function. Using laser-evoked potentials (LEP), the authors sought information on small afferent (nociceptive) function. METHODS: The brain potentials evoked by CO(2) laser pulses directed to the perioral and supraorbital regions were studied in 67 patients with idiopathic or symptomatic trigeminal neuralgia and 30 normal subjects. Of the 67 patients, 49 were receiving carbamazepine. RESULTS: All patients with symptomatic and 51% of those with idiopathic trigeminal neuralgia had frankly abnormal LEP on the painful side. The mean latency was significantly higher and mean amplitude lower on the painful than the nonpainful side. However, even on the nonpainful side, the mean latency was significantly longer than that of the age-matched controls. The nonpainful-side latency correlated significantly with the carbamazepine dose. CONCLUSIONS: LEP detect severe impairment of the nociceptive afferent system on the painful side of patients with idiopathic as well as symptomatic trigeminal neuralgia. A dysfunction of small-myelinated afferents may play an important role in the pathophysiology of neuralgic pain. Carbamazepine markedly dampens these brain potentials. The authors propose that this effect may result from inhibition of nociceptive transmission in the cingulate gyrus. PMID- 11425941 TI - Painful alcoholic polyneuropathy with predominant small-fiber loss and normal thiamine status. AB - BACKGROUND: Although polyneuropathy related to chronic alcoholism has been reported frequently, its clinical features and pathogenesis remain to be clarified. OBJECTIVE: To determine the clinicopathologic features and pathogenesis of alcoholic polyneuropathy associated with pain in patients with normal thiamine status, particularly in comparison to beriberi neuropathy. PATIENTS AND METHODS: Clinical, electrophysiologic, and histopathologic findings were assessed in 18 patients with painful alcoholic polyneuropathy and normal thiamine status. RESULTS: Symmetric sensory-dominant polyneuropathy predominantly involving the lower limbs was the major clinical pattern. Painful sensations with or without burning quality represented the initial and major symptom. Progression of symptoms usually was gradual, continuing over months or years. Electrophysiologic and pathologic findings mainly indicated an axonal neuropathy. Densities of small myelinated fibers and unmyelinated fibers were more severely reduced than the density of large myelinated fibers, except in patients with a long history of neuropathic symptoms and marked axonal sprouting. CONCLUSIONS: The clinicopathologic features of painful symptoms and small axon loss are distinct from those of beriberi neuropathy. Sensory-dominant involvement with prominent neuropathic pain is characteristic of alcoholic neuropathy when thiamine deficiency is not involved, supporting the view of direct neurotoxic effect by alcohol or its metabolites. PMID- 11425942 TI - A non-ischemic forearm exercise test for the screening of patients with exercise intolerance. AB - BACKGROUND: The forearm exercise test is a common investigation that allows detection of some metabolic myopathies. It is not completely standardized and, when performed in ischemic conditions, may induce rhabdomyolysis in patients with glycogenosis. OBJECTIVE: To develop a standardized non-ischemic exercise test for a safe screening of patients with exercise intolerance. METHODS: Twenty-six healthy subjects and 32 patients with exercise intolerance performed an isometric exercise at 70% of the maximal voluntary contraction during 30 seconds in non ischemic conditions. Blood concentrations of creatine kinase, lactate, and ammonia were analyzed. RESULTS: A nearly fourfold lactate rise was induced by exercise in healthy subjects. All patients with normal muscle biopsy showed values similar to those of healthy subjects. No significant lactate increase was observed in six patients with a myophosphorylase defect and one with a debrancher defect. Disparate lactate responses were observed in 14 patients with a mitochondrial myopathy. The blood lactate level at rest was abnormally high in four of these patients. The lactate surface normalized by the mechanical energy production was above the normal range in eight patients. CONCLUSIONS: The authors propose a standardized non-ischemic grip test that overcomes the main drawbacks of the classic ischemic forearm exercise test. It provides a specific, efficient, and safe screening test for patients with exercise intolerance. Its sensitivity was very good for patients with a glycogenolysis defect but remains partial in patients with a mitochondrial disorder. PMID- 11425943 TI - Surprises of genetic engineering: a possible model of polyglucosan body disease. AB - BACKGROUND: The authors previously reported the generation of a knockout mouse model of Pompe disease caused by the inherited deficiency of lysosomal acid alpha glucosidase (GAA). The disorder in the knockout mice (GAA-/-) resembles the human disease closely, except that the clinical symptoms develop late relative to the lifespan of the animals. In an attempt to accelerate the course of the disease in the knockouts, the authors increased the level of cytoplasmic glycogen by overexpressing glycogen synthase (GSase) or GlutI glucose transporter. METHODS: GAA-/- mice were crossed to transgenic mice overexpressing GSase or GlutI in skeletal muscle. RESULTS: Both transgenics on a GAA knockout background (GS/GAA-/ and GlutI/GAA-/-) developed a severe muscle wasting disorder with an early age at onset. This finding, however, is not the major focus of the study. Unexpectedly, the mice bearing the GSase transgene, but not those bearing the GlutI transgene, accumulated structurally abnormal polysaccharide (polyglucosan) similar to that observed in patients with Lafora disease, glycogenosis type IV, and glycogenosis type VII. Ultrastructurally, the periodic acid-Schiff (PAS) positive polysaccharide inclusions were composed of short, amorphous, irregular branching filaments indistinguishable from classic polyglucosan bodies. The authors show here that increased level of GSase in the presence of normal glycogen branching enzyme (GBE) activity leads to polyglucosan accumulation. The authors have further shown that inactivation of lysosomal acid alpha-glucosidase in the knockout mice does not contribute to the process of polyglucosan formation. CONCLUSIONS: An imbalance between GSase and GBE activities is proposed as the mechanism involved in the production of polyglucosan bodies. The authors may have inadvertently created a "muscle polyglucosan disease" by simulating the mechanism for polyglucosan formation. PMID- 11425944 TI - The Monro-Kellie hypothesis: applications in CSF volume depletion. AB - More than two centuries ago, Alexander Monro applied some of the principles of physics to the intracranial contents and for the first time hypothesized that the blood circulating in the cranium was of constant volume at all times. This hypothesis was supported by experiments by Kellie. In its original form, the hypothesis had shortcomings that prompted modification by others. What finally came to be known as the Monro-Kellie doctrine, or hypothesis, is that the sum of volumes of brain, CSF, and intracranial blood is constant. An increase in one should cause a decrease in one or both of the remaining two. This hypothesis has substantial theoretical implications in increased intracranial pressure and in decreased CSF volume. Many of the MRI abnormalities seen in intracranial hypotension or CSF volume depletion can be explained by the Monro-Kellie hypothesis. These abnormalities include meningeal enhancement, subdural fluid collections, engorgement of cerebral venous sinuses, prominence of the spinal epidural venous plexus, and enlargement of the pituitary gland. PMID- 11425945 TI - Hand weakness from a precentral gyrus infarct with intermittent hypotension. PMID- 11425946 TI - Normal plasma levels of orexin A (hypocretin-1) in narcoleptic patients. AB - Deficient orexin signaling has been shown to cause narcolepsy-like conditions in animals. In human narcolepsy, CSF levels of orexin A (hypocretin-1) were reported to be low in most cases. The authors measured CSF and plasma orexin A levels in patients with narcolepsy and in controls. Confirming earlier studies, they found CSF orexin A levels to be extremely low in patients with narcolepsy. However, plasma orexin A levels did not differ from those observed in controls. These results suggest that orexin deficiency in patients with narcolepsy is a phenomena restricted to the CNS. PMID- 11425947 TI - Narcolepsy and low CSF orexin (hypocretin) concentration after a diencephalic stroke. AB - Idiopathic narcolepsy usually results from a loss of the hypothalamic neuropeptide orexin (hypocretin), but the cause of secondary narcolepsy resulting from focal brain lesions is unknown. The authors describe a young man who developed narcolepsy after a large hypothalamic stroke. His lesion included much of the hypothalamic region in which orexin is produced, and his CSF concentration of orexin was low. The authors hypothesize that a loss of orexin neurons or their relevant targets may be the specific neuropathology causing this and many other cases of secondary narcolepsy. PMID- 11425948 TI - Cytoplasmic and nuclear polyglutamine aggregates in SCA6 Purkinje cells. AB - Aggregations of the alpha1A-calcium channel protein have been previously demonstrated in spinocerebellar ataxia type 6 (SCA6). Here the authors show that small aggregates, labeled by a monoclonal antibody 1C2 that preferentially detects expanded polyglutamine larger than that in SCA6 mutation, are present mainly in the cytoplasm but also in the nucleus of Purkinje cells. Although the length of expansion is small in SCA6, the current finding might indicate that SCA6 conforms to the pathogenic mechanism(s) in other polyglutamine diseases. PMID- 11425949 TI - Association study of dopamine D2, D3 receptor gene polymorphisms with motor fluctuations in PD. AB - The authors investigated the association between dopamine receptor D2, D3 gene polymorphisms, and the risk of developing motor fluctuations in PD. DRD3 BalI and MspI polymorphisms were not associated with risk of developing motor fluctuations. However, the genotypic distribution of DRD2 TaqIA polymorphism was significantly different in motor fluctuators and nonmotor fluctuators. These findings suggest that DRD2 TaqIA polymorphism may be associated with an increased risk for developing motor fluctuations in PD. PMID- 11425950 TI - The nigrostriatal dopaminergic system in familial early onset parkinsonism with parkin mutations. AB - Nigrostriatal dopaminergic function and cerebral energy metabolism were measured with PET in two brothers with early-onset parkinsonism caused by mutation of the parkin gene. Energy metabolism did not differ, but the nigrostriatal dopaminergic pattern was clearly different than that of sporadic PD. Thus parkinsonism in these two patients was shown to be pathophysiologically different than PD. PMID- 11425951 TI - Clinical aspects of multifocal or generalized tonic dystonia in reflex sympathetic dystrophy. AB - The authors describe 10 patients with reflex sympathetic dystrophy that progressed to a multifocal or generalized tonic dystonia. The neuropsychologic profile was similar to that of other patients with chronic pain, irrespective of its cause. The distribution pattern of dystonia, the stretch reflex abnormalities, and the worsening of dystonia after tactile and auditory stimuli suggest impairment of interneuronal circuits at the brainstem or spinal level. Antibody titers for glutamic acid decarboxylase, tetanus, and Sjogren antigens were all normal. PMID- 11425952 TI - Mononeuropathy multiplex as the initial manifestation of neurofibromatosis type 2. AB - The authors report a patient with neurofibromatosis type 2 (NF2) presenting with an axonal mononeuropathy multiplex. Sural nerve biopsy showed small scattered groups of Schwann cells transformed into irregular branching cells with abnormal cell-cell contacts. The authors hypothesize that defective Schwann cell function, due to inactivation of the NF2 gene product merlin, leads to changes in morphology, cell-cell contact, and growth, and finally to degeneration of axons. PMID- 11425953 TI - Perilymph fistula associated with pulse-synchronous eye oscillations. AB - Three-dimensional eye movements (scleral search coil system) were recorded in a patient with a surgically acquired perilymph fistula of the left horizontal semicircular canal. Spontaneous horizontal pendular nystagmus was found to be related to the heart rate and may be caused by pressure transfer of blood pulses to the labyrinth. In addition, a contralesional horizontal jerk nystagmus was elicited by Valsalva maneuver, indicating that Ewald's first law may not only be valid for excitation but also for inhibition. PMID- 11425954 TI - Long-term continuation of levetiracetam in patients with refractory epilepsy. AB - The long-term continuation (retention) rate, efficacy, and safety data of the new antiepileptic drug levetiracetam (LEV) was evaluated in all patients with epilepsy exposed to the drug during its developmental program (n = 1,422). The retention rate was estimated to be 60% after 1 year and 32% after 5 years. Thirty nine percent (512/1,325) of patients had a seizure reduction of > or =50%, and 13% (183/1,422) became seizure-free for at least 6 months. LEV seems an effective and well tolerated new antiepileptic drug. PMID- 11425955 TI - A hypersomnolent girl with decreased CSF hypocretin level after removal of a hypothalamic tumor. PMID- 11425956 TI - Very late onset Friedreich's presenting as spastic tetraparesis without ataxia or neuropathy. PMID- 11425957 TI - Detection of enteroviral sequences from frozen spinal cord samples of Japanese ALS patients. PMID- 11425958 TI - Encephalitis associated with Chlamydia pneumoniae. PMID- 11425959 TI - Acute compartment syndrome after forearm ischemic work test in a patient with McArdle's disease. PMID- 11425960 TI - The value of informant versus individual's complaints of memory impairment in early dementia. PMID- 11425961 TI - Olanzapine for psychosis in Parkinson's disease. PMID- 11425962 TI - Competency to consent to medical treatment in cognitively impaired patients with Parkinson's disease. PMID- 11425963 TI - PCV chemotherapy for recurrent glioblastoma multiforme. PMID- 11425964 TI - Focus on imaging at the American Society of Nephrology annual meeting. PMID- 11425965 TI - Pancreatic carcinoma: developing a protocol for multi-detector row CT. PMID- 11425967 TI - Coding and reimbursement issues for the radiologist. AB - Radiologists are dependent on the proper execution of a complex administrative chain of disease and diagnosis coding and procedural coding to be properly reimbursed for the examinations they perform. The radiologist bears the ultimate responsibility for the appropriateness of these codes. However, many practicing radiologists are unaware of the critical link between the two coding systems and the systems that have developed to provide a common method of describing diseases, diagnoses, and procedures. This article is an introduction to these systems, and it emphasizes the importance of the involvement of the radiologist in coding. The inherent complexity of the reimbursement system in use is emphasized, as well as the essential role of the radiologist in complying with these often complicated and ever-changing directives. PMID- 11425968 TI - MR imaging of the breast for the detection, diagnosis, and staging of breast cancer. AB - With the introduction of contrast agents, advances in surface coil technology, and development of new imaging protocols, contrast agent-enhanced magnetic resonance (MR) imaging has emerged as a promising modality for detection, diagnosis, and staging of breast cancer. The reported sensitivity of MR imaging for the visualization of invasive cancer has approached 100%. There are many examples in the literature of MR imaging--demonstrated mammographically, sonographically, and clinically occult breast cancer. Often, breast cancer detected on MR images has resulted in a change in patient care. Despite these results, there are many unresolved issues, including no defined standard technique for contrast-enhanced breast MR imaging, no standard interpretation criteria for evaluating such studies, no consensus on what constitutes clinically important enhancement, and no clearly defined clinical indications for the use of MR imaging. Furthermore, this technology remains costly, and issues of cost effectiveness and cost competition from percutaneous biopsy have yet to be fully addressed. These factors along with the lack of commercially available MR imaging -guided localization and biopsy systems have slowed the transfer of this imaging technology from research centers to clinical breast imaging practices. Technical requirements, potential clinical applications, and potential pitfalls and limitations of contrast-enhanced MR imaging as a method to help detect, diagnose, and stage breast cancer will be described. PMID- 11425969 TI - MR imaging--guided large-core (14-gauge) needle biopsy of small lesions visible at breast MR imaging alone. AB - PURPOSE: To report our experience with magnetic resonance (MR) imaging-guided large-core breast biopsy of lesions visible at breast MR imaging only. MATERIALS AND METHODS: Stereotactic large-core (14-gauge) needle biopsy of 78 lesions visible at MR imaging only was performed with MR imaging guidance in 59 patients. Results were validated with excisional biopsy or mastectomy in 42 lesions and with radiologic-pathologic correlation and/or follow-up MR imaging for at least 2 years in another 17 lesions. The accuracy of MR imaging--guided core biopsy was determined for those 59 lesions with established validation. The effect on patient treatment was evaluated by comparing the prebiopsy treatment plan with the ultimate treatment. RESULTS: Histologic diagnosis from core biopsy was possible in 77 (99%) of 78 lesions. In the 59 lesions with established validation, the diagnostic accuracy of MR imaging--guided core biopsy was 98% (58 of 59). Successful MR imaging--guided core biopsy findings changed treatment in 70% (54 of 77) of lesions. Difficulties were due to the unsatisfactory performance of earlier types of MR imaging--compatible biopsy guns and decreasing target visibility during intervention. CONCLUSION: MR imaging--guided large-core stereotactic breast biopsy is sufficiently accurate for obtaining histologic proof of lesions visible only at MR imaging. It can change patient treatment by reducing unnecessary surgical biopsy and can enable one-step surgery for breast cancers. PMID- 11425970 TI - Human breast lesions: characterization with contrast-enhanced in vivo proton MR spectroscopy--initial results. AB - PURPOSE: To assess the clinical usefulness of localized proton (hydrogen 1) magnetic resonance (MR) spectroscopy in the characterization of contrast material enhanced breast lesions on the basis of choline detection. MATERIALS AND METHODS: Examinations were performed at 1.5 T with use of a standard breast coil. Contrast enhanced MR imaging was performed in 30 consecutive patients (mean age, 50 years; age range, 20--80 years) who had nonspecific lesions (>1.5 cm in diameter) on sonograms or mammograms. Single-voxel (1)H MR spectroscopy was performed in the enhancing lesions by using a point-resolved spectroscopic sequence with echo times of 38, 135, and 270 msec. MR spectroscopic and histopathologic findings were determined in blinded fashion and compared. RESULTS: Twenty-four patients had carcinoma of the breast (tumor size, 2.0--11.2 cm; mean, 4.7 cm), and six had benign lesions (lesion size, 1.8--3.8 cm; mean, 2.7 cm). Choline was detected in 22 patients with carcinoma. Choline was not detected in five patients with benign lesions and in two patients with carcinoma. The preliminary results indicate that this technique had a sensitivity of 92%, specificity of 83%, and accuracy of 90%. CONCLUSION: Choline can be reliably detected in less than 45 minutes in large contrast-enhanced breast lesions by using a multiecho point-resolved spectroscopic protocol. The presence of water-soluble choline metabolites obtainable with (1)H MR spectroscopy could complement MR imaging findings to improve specificity and to reduce the number of unnecessary biopsies. PMID- 11425971 TI - Breast cancer: variables affecting sentinel lymph node visualization at preoperative lymphoscintigraphy. AB - PURPOSE: To compare patients with visualized sentinel lymph nodes (SLNs) and patients with nonvisualized SLNs, with a focus on variables affecting SLN visualization at preoperative lymphoscintigraphy and on nodal drainage basins as related to tumor location. MATERIALS AND METHODS: One hundred thirty-six patients who had breast cancer underwent preoperative lymphoscintigraphy before SLN biopsy. Patients with visualized and nonvisualized SLNs were compared for age; tumor site, size, and histologic findings; injection guidance method; diagnostic biopsy type; interval between biopsy and lymphoscintigraphy; intraoperative identification method; and surgical identification rate. Visualized SLN drainage basins were noted. RESULTS: Ninety-nine patients had visualized and 37 had nonvisualized SLNs, without statistically significant differences in tumor site, size, and histologic findings; injection guidance method; diagnostic biopsy type; and interval between biopsy and lymphoscintigraphy. Ninety-nine (73%) of the 136 SLNs were visualized at lymphoscintigraphy; 30 (81%) of the 37 nonvisualized SLNS were identified at surgery. Of the seven SLNs not identified at surgery, five were mapped with radiocolloid only. Patients with nonvisualized SLNs were older than those with visualized SLNs. Eleven (46%) of 24 tumors with internal mammary drainage were in the outer part of the breast. CONCLUSION: Patients with and those without visualization differed in age, SLN identification at surgery, and surgical identification method. Nonvisualized status does not preclude axillary metastasis. In older patients with nonvisualized SLNs, blue dye may aid in SLN detection, as compared with isotope-only localization. PMID- 11425972 TI - Detection of malignant tumors: whole-body PET with fluorine 18 alpha-methyl tyrosine versus FDG--preliminary study. AB - PURPOSE: To compare the diagnostic potential of whole-body positron emission tomography (PET) with fluorine 18 alpha-methyl tyrosine (FMT) with that of whole body PET with 2-[fluorine 18]fluoro-2-deoxy-D-glucose (FDG). MATERIALS AND METHODS: Nineteen patients with or suspected of having malignant tumors and five healthy volunteers underwent whole-body PET with FMT and FDG. RESULTS: In comparison with FDG uptake, FMT uptake was significantly less in the brain, heart, lung, liver, and spine. On a lesion-by-lesion basis, the sensitivity of whole-body FMT PET for depicting malignant tumors was inferior to that of whole body FDG PET, but this difference was not statistically significant (74% [26 of 35 lesions] vs 91% [32 of 35 lesions], P >.05). The positive predictive value of FMT PET was superior to that of FDG PET (87% [26 of 30 lesions] vs 63% [32 of 51 lesions], P <.001). The difference in uptake between benign and malignant lesions was significant with FMT PET (mean +/- SD, 1.64 +/- 0.96 vs 0.79 +/- 0.23; P <.001) but not with FDG PET (5.02 +/- 3.56 vs 4.02 +/- 2.90, P >.05). CONCLUSION: Whole-body FMT PET is clinically useful in the diagnosis of malignant tumors and may be effective in the depiction of primary and metastatic lesions in the cardiac region or in the brain. PMID- 11425973 TI - Congenital diaphragmatic hernia: prenatal evaluation with MR lung volumetry- preliminary experience. AB - PURPOSE: To determine the interobserver variability of prenatal magnetic resonance (MR) lung volumetry and to assess the value of MR lung volumetric findings as predictors of outcome in fetuses with congenital diaphragmatic hernia. MATERIALS AND METHODS: Prenatal MR imaging was performed in 26 fetuses with unilateral congenital diaphragmatic hernia. Two independent observers performed planimetric measurement of lung volume. Relative lung volume was calculated as the observed total lung volume expressed as a percentage of the total lung volume predicted from fetal size. Relative lung volume was correlated with the ultrasonographic lung-head ratio in left-sided congenital diaphragmatic hernias evaluated before 27 weeks gestation (n = 21) and with pregnancy outcome in all cases of isolated left-sided congenital diaphragmatic hernia without prenatal intervention (n = 11). RESULTS: Observers demonstrated excellent agreement in total lung volume measurements at MR imaging, with an intraclass correlation coefficient of 0.95. Relative lung volume was positively correlated with lung-head ratio (r = 0.78, P <.001). By using rank order analysis in the pregnancy outcome group, relative lung volume was predictive of prognosis (P <.05) when adjusted for gestational age at delivery and birth weight. Three of four fetuses with a relative lung volume of less than 40% died. CONCLUSION: Interobserver agreement is high at MR lung volumetry, and its findings are predictive of outcome in fetuses with isolated left-sided congenital diaphragmatic hernia. PMID- 11425974 TI - The twin peak sign. PMID- 11425975 TI - Testicular microlithiasis: what is its association with testicular cancer? AB - PURPOSE: To determine the prevalence of testicular microlithiasis in patients who were referred for scrotal ultrasonography (US) at a tertiary care cancer center and to evaluate the association between microlithiasis and cancer. MATERIALS AND METHODS: Testicular sonograms obtained in 528 men were retrospectively reviewed to identify patients with US findings suggestive of microlithiasis, intratesticular masses, and intratesticular heterogeneous changes. The association of US findings with medical records and with histopathologic findings that were available in 95 patients was evaluated. Statistical analysis was performed to determine the relationship of testicular cancer, intratesticular mass, and microlithiasis. RESULTS: Forty-eight (9%) of the 528 patients had microlithiasis; 13 of these (27%) had testicular cancers. Of the 480 patients without microlithiasis, 38 (8%) had testicular cancer. Ninety patients had an intratesticular mass, of whom 23 (26%) had microlithiasis. Forty-three (12 with microlithiasis) patients with a mass had testicular cancer, 43 (10 with microlithiasis) had benign findings or nontesticular malignant histopathologic findings, and four (one with microlithiasis) had no pathologic findings. CONCLUSION: Intratesticular microlithiasis is highly associated with confirmed testicular cancer, as well as with US evidence of testicular mass. PMID- 11425976 TI - Abdominopelvic actinomycosis involving the gastrointestinal tract: CT features. AB - PURPOSE: To assess the computed tomographic (CT) features of abdominopelvic actinomycosis involving the gastrointestinal tract. MATERIALS AND METHODS: CT scans were analyzed in 18 patients with pathologically proved abdominopelvic actinomycosis involving the gastrointestinal tract. Eight patients had a history of using intrauterine contraceptive devices. Bowel site, wall thickness, length, bowel involvement patterns, inflammatory infiltration, and features of peritoneal or pelvic mass, if present, were evaluated at CT. RESULTS: Of the gastrointestinal tract, the sigmoid colon was most commonly involved (50%). All patients showed concentric (n = 15) or eccentric (n = 3) bowel wall thickening, with a mean thickness of 1.2 cm and a mean length of 8.3 cm. The thickened bowel enhanced homogeneously in nine patients and heterogeneously in the other nine. Inflammatory infiltration was mostly diffuse and severe. In 17 patients, a peritoneal or pelvic mass (mean maximum diameter, 3.2 cm) was seen adjacent to the involved bowel and appeared to be heterogeneously enhanced in most cases; infiltration into the abdominal wall was seen in four patients. CONCLUSION: Actinomycosis should be included in the differential diagnosis when CT scans show bowel wall thickening and regional pelvic or peritoneal mass with extensive infiltration, especially in patients with abdominal pain, fever, leukocytosis, or long-term use of intrauterine contraceptive devices. PMID- 11425977 TI - Measurement of anal sphincter muscles: endoanal US, endoanal MR imaging, or phased-array MR imaging? A study with healthy volunteers. AB - PURPOSE: To compare endoanal ultrasonography (US), endoanal magnetic resonance (MR) imaging, and phased-array MR imaging for anal sphincter muscle measurement. MATERIALS AND METHODS: Sixty healthy volunteers underwent 1.5-T phased-array MR, endoanal MR, and endoanal US examinations. Sphincter muscle thicknesses were measured. Measurement reliability was analyzed, and correlations among the imaging methods were calculated. Multivariate analysis was performed to assess the influence of age, weight, height, sex, parity, and obstetric trauma on sphincter dimensions. RESULTS: Both MR methods had good reliability for measurements of all sphincter components, whereas endoanal US was reliable for internal sphincter measurement only. There was little correlation between the techniques, except between the two MR techniques, with a strong correlation for total sphincter and perineal body thickness. The internal sphincter thickened significantly (P =.002) with age at endoanal US and endoanal MR imaging but not at phased-array MR imaging. There were small sex-based differences in sphincter muscle measurements at phased-array MR imaging only. CONCLUSION: Endoanal US enables reliable measurement of only internal sphincter thickness, whereas both MR imaging methods enable reliable measurement of all sphincter components. Sphincter measurement with phased-array MR imaging is as reliable as that with endoanal MR imaging. PMID- 11425978 TI - Malignant hilar and perihilar biliary obstruction: use of MR cholangiography to define the extent of biliary ductal involvement and plan percutaneous interventions. AB - PURPOSE: To determine the usefulness of magnetic resonance (MR) cholangiography in defining the extent of biliary ductal involvement in patients with malignant hilar and perihilar biliary obstruction and to evaluate whether findings at MR cholangiography alone are sufficient to plan percutaneous interventions in these patients. MATERIALS AND METHODS: Twenty-nine patients with malignant hilar and perihilar biliary obstruction were examined with MR cholangiography. Two radiologists evaluated MR images and determined the extent of biliary ductal involvement. A hypothetical plan for biliary drainage was established prior to any intervention. All patients underwent percutaneous cholangiography, and 27 of 29 patients also underwent biliary drainage and/or stent placement within 7 days after MR cholangiography. By using direct cholangiography as the standard of reference, the usefulness of MR cholangiography in defining the extent of biliary ductal involvement was determined. The type of drainage performed was compared with the type that had been anticipated at MR cholangiography. RESULTS: MR cholangiography was adequate in helping predict the extent of biliary ductal involvement in 28 (96%) of 29 patients and led to underestimation of the extent of the disease in one patient. The therapeutic plan anticipated with MR cholangiography matched the one actually used in 24 (83%) of 29 patients. CONCLUSION: The high accuracy of MR cholangiography for defining extent of ductal involvement in patients with malignant hilar and perihilar obstruction allows adequate planning of percutaneous interventions in a majority of patients. PMID- 11425979 TI - Multi--detector row helical CT of the pancreas: effect of contrast-enhanced multiphasic imaging on enhancement of the pancreas, peripancreatic vasculature, and pancreatic adenocarcinoma. AB - PURPOSE: To determine the optimal phase for enhancement of the normal pancreas and peripancreatic vasculature and the maximal tumor-to-pancreatic parenchymal enhancement difference by using multiphase, contrast material-enhanced, multi detector row helical computed tomography (CT). MATERIALS AND METHODS: Forty-nine patients with a normal-appearing pancreas but suspected of having pancreatic abnormality and 28 patients with proved pancreatic adenocarcinoma underwent multiphase, contrast-enhanced, multi-detector row CT during the arterial phase (AP), pancreatic parenchymal phase (PPP), and portal venous phase (PVP). Attenuation values of the normal pancreas, pancreatic adenocarcinoma, celiac and superior mesenteric arteries, and superior mesenteric and portal veins were measured during all three imaging phases. Quantitative analysis of these measurements and subjective qualitative analysis of tumor conspicuity were performed. RESULTS: Maximal enhancement of the normal pancreatic parenchyma occurred during the PPP. Maximal tumor-to-parenchyma attenuation differences during the PPP and PVP were equivalent but greater than that during the AP. Subjective analysis revealed that tumor conspicuity during the PPP and PVP was equivalent but superior to that during the AP. Maximal arterial enhancement was seen during the PPP, and maximal venous enhancement was seen during the PVP. CONCLUSION: A combination of PPP and PVP imaging is sufficient for detection of pancreatic adenocarcinoma, because it provides maximal pancreatic parenchymal and peripancreatic vascular enhancement. AP imaging can be reserved for patients in whom CT angiography is required. PMID- 11425980 TI - Effect of cross-sectional imaging on negative appendectomy and perforation rates in children. AB - PURPOSE: To compare negative appendectomy and perforation rates in children who underwent ultrasonography (US), computed tomography (CT), or no imaging before urgent appendectomy. MATERIALS AND METHODS: All children who underwent urgent appendectomy during a 4(1/2)-year period were identified in a surgical billing database. Pathology reports were coded as negative or as showing acute inflammation or perforation. Imaging up to 14 days before appendectomy or abscess drainage was noted, and imaging-based diagnoses were compared with pathologic findings. Patient age and sex were recorded. RESULTS: Two hundred ninety-nine children, 176 (59%) male and 123 (41%) female (mean age, 10.4 years; age range, 1 -21 years), underwent urgent appendectomy. One hundred twenty-six (42%) underwent no imaging, 121 (41%) underwent US with or without CT, and 52 (17%) underwent CT only; 44 (15%) underwent both US and CT. There were significantly higher rates of appendectomy with normal pathologic findings ("negative appendectomy") in patients who underwent no imaging (14% [18 of 126]) or US (17% [20 of 121]) versus the rates in those who underwent CT only (2% [one of 52]) (P =.02 and P =.007, respectively). The negative appendectomy rate was 7% in 96 patients who underwent CT with or without prior US. The perforation rates were not significantly different. CONCLUSION: As compared with children who underwent no preoperative imaging and those who underwent US, children who underwent CT had a significantly lower negative appendectomy rate, without a significantly higher perforation rate. PMID- 11425981 TI - Pediatric radiology at the millennium. AB - PURPOSE: To evaluate the current practice of and job market for pediatric radiologists in the United States and Canada with a 1998 survey and compare findings with those of surveys from 1980 and 1989. MATERIALS AND METHODS: Surveys were mailed to the 728 active members of the Society for Pediatric Radiology. Questions covered professional practice, subspecialization, and involvement in evolving technologies. A pediatric radiology help-wanted index was generated from a diagnostic radiology help-wanted index. RESULTS: Increasing involvement in the interpretation of computed tomographic, ultrasonographic, and magnetic resonance images was found among the 57% (411 of 728) of the members who responded. The attainment of a certificate of added qualification in pediatric radiology was found in nearly three-fourths of the membership, and 85% (348 of 408) had completed a fellowship. More than half of the respondents were involved in interventional procedures amid a continued increase in volume; 24% (100 of 409) of the membership, as compared with 7% in the 1980 survey, was practicing in a community or "other" setting. Subspecialization within pediatric radiology had diminished, and there was a larger percentage of female pediatric radiologists, particularly among the newest members. The job market was robust, having recovered substantially since the middle 1990s. CONCLUSION: The practice of pediatric radiology has evolved, with increasing involvement in advanced techniques and modalities, as well as a greater presence in community settings. The help-wanted index supports the recent discussions of a personnel shortfall. PMID- 11425982 TI - Early gastric carcinoma: evaluation with high-spatial-resolution MR imaging in vitro. AB - PURPOSE: To determine signal intensity characteristics of the gastric wall layers and to assess the accuracy of the evaluation of early gastric carcinomas in vitro by using resected specimens studied with high-spatial-resolution magnetic resonance (MR) imaging. MATERIALS AND METHODS: Fifteen gastric specimens obtained from patients suspected of having early gastric carcinoma were studied with a 1.5 T MR system with a 4-cm-diameter loop coil. High-spatial-resolution spin-echo MR images were obtained with a field of view of 50 mm, a matrix of 256 x 256, and a section thickness of 2 mm, resulting in a voxel size of 0.08 mm(3). Findings from MR images were compared with histopathologic findings. RESULTS: T1- and T2 weighted MR images clearly depicted the normal gastric wall as consisting of four and six layers, respectively, which corresponded well to the histologic layers. In 14 (93%) of 15 gastric carcinomas, the depth of mural invasion visualized with MR imaging correlated well with the histopathologic stage. The stage determined with MR imaging, however, was lower in one instance (7%) than the histopathologic stage. MR imaging also depicted the gross features of the tumor, presence of ulceration, and adjacent lymph node swelling. CONCLUSION: High-spatial-resolution MR imaging has a high diagnostic accuracy in the evaluation of the mural invasion of early gastric carcinoma in vitro and thus potentially enables preoperative histopathologic staging. PMID- 11425983 TI - Cryptococcomas distinguished from gliomas with MR spectroscopy: an experimental rat and cell culture study. AB - PURPOSE: To use magnetic resonance (MR) spectroscopy to characterize clinical isolates of Cryptococcus neoformans and a glioma cell line in culture and in experimental rats. MATERIALS AND METHODS: One- and two-dimensional hydrogen 1 MR spectra were acquired from fungi cultured in vitro (16 isolates of C neoformans, three of Candida albicans, three of Aspergillus fumigatus, three of Saccharomyces cerevisiae) and a C6 glioma cell line. Cerebral biopsy specimens were obtained from healthy rats and animals with experimental infections or gliomas (19 healthy brains, 20 cryptococcomas, and 19 gliomas). Unequivocal signal assignment was performed for cell suspensions and tissue samples by using homo- and heteronuclear two-dimensional correlation spectra. RESULTS: MR spectra of C neoformans and cerebral cryptococcomas--but not of other fungi, healthy brains, or gliomas--were dominated by resonances from the cytosolic disaccharide alpha,alpha-trehalose. This spectral pattern was different from that of gliomas, which was dominated by lipids and an increased choline-creatine ratio, and that of healthy brain. CONCLUSION: A remarkably high concentration of alpha,alpha trehalose in relation to other metabolites that are visible with MR spectroscopy is diagnostic of C neoformans. Cerebral cryptococcomas are an uncommon but serious manifestation of cryptococcosis in humans. Application of these results to the noninvasive diagnosis of cerebral cryptococcomas would help reduce the risk and expense of unnecessary surgery or biopsy and expedite patient treatment. PMID- 11425984 TI - Interstitial T1-weighted MR lymphography: lipophilic perfluorinated gadolinium chelates in pigs. AB - PURPOSE: To investigate the enhancement of the regional lymph nodes, lymphatic vessels, and thoracic duct after interstitial administration of lymphotropic perfluorinated gadolinium chelates at magnetic resonance (MR) imaging. MATERIALS AND METHODS: Two perfluorinated gadolinium chelates, gadofluoramide and gadofluorine 8, were injected subcutaneously into the hind legs of 10 pigs, respectively. Both contrast media were studied at doses of 10 and 25 micromol per kilogram of body weight. T1-weighted three-dimensional gradient-echo and maximum intensity projection images were obtained at 1.5 T between 1 and 210 minutes and 24 hours after injection. The contrast agents were qualitatively compared regarding enhancement and depiction of the regional lymph nodes, lymphatic vessels, and thoracic duct. RESULTS: The inguinal and iliac lymph nodes and lymphatic vasculature enhanced substantially within 10 minutes after subcutaneous administration of both lymphotropic contrast agents. Gadofluorine 8 showed a lymphographic effect superior to that of gadofluoramide. The paraaortic lymph nodes and thoracic duct were best visualized 10--50 minutes after injection of 25 micromol/kg of gadofluorine 8. Lymphatic system enhancement diminished after 2 hours, and the liver and bowel tract enhanced within 24 hours. CONCLUSION: Interstitial administration of perfluorinated gadolinium chelates offers great potential for T1-weighted MR lymphography with positive enhancement of the lymph nodes and lymphatic vasculature. PMID- 11425985 TI - Prototype percutaneous thrombolytic device: preclinical testing in subacute inferior vena caval thrombosis in a pig model. AB - PURPOSE: To develop an animal model of subacute inferior vena caval (IVC) thrombosis and apply this model in evaluating the safety and efficacy of a prototype percutaneous thrombolytic device for restoring patency. MATERIALS AND METHODS: In 11 pigs, a stent with a ligature in the middle was placed in the IVC. Thrombin was injected to induce thrombosis. Hemostasis was achieved by using an occlusion balloon. The stent was ligated to prevent thrombus migration. Five to 8 days after thrombus induction, the ligature was broken and the stent fully deployed. In 10 animals, thrombectomy was performed by using the percutaneous thrombolytic device. A vena caval filter was inserted at the beginning of each declotting procedure. Thrombus removal percentage was estimated and pulmonary angiograms obtained to detect embolism before and after thrombectomy. The IVC was analyzed histologically. To determine thrombus composition, one animal was sacrificed without thrombectomy. Concerning procedural safety, failure of the stent delivery system, stent migration, and venous perforation due to balloon inflation and the stent placement or thrombectomy procedure were evaluated. RESULTS: Thrombus creation was successful in all animals. Fragmentation led to 75%--100% thrombus removal with flow restoration in all cases. There were no episodes of stent delivery failure, stent migration, or venous perforation. No significant pulmonary embolism was observed. In one case, a vessel dissection was identified at histologic examination. CONCLUSION: In this animal model of IVC thrombosis, the percutaneous thrombolytic device is effective and safe for clot removal. PMID- 11425986 TI - Fatal late coronary thrombosis after implantation of a radioactive stent: postmortem angiographic and histologic findings--case report. AB - Postmortem angiography and histologic analysis of a fatal coronary thrombosis 4 months after implantation of a radioactive stent are described. Histologic findings suggested incomplete re-endothelialization in the segment with the stent. Ionizing radiation may delay re-endothelialization after revascularization, thus maintaining the thrombogenicity of the irradiated vessel segment. Thus, prolonged antiplatelet therapy should be considered after intravascular radiation therapy. PMID- 11425987 TI - Percutaneous radio-frequency ablation of liver metastases from breast cancer: initial experience in 24 patients. AB - PURPOSE: To evaluate the authors' initial experience in a consecutive series of 24 patients with breast cancer liver metastases treated with radio-frequency (RF) ablation. MATERIALS AND METHODS: Twenty-four consecutive patients with 64 metastases measuring 1.0--6.6 cm in diameter (mean, 1.9 cm) underwent ultrasonography-guided percutaneous RF ablation with 18-gauge, internally cooled electrodes. Treatment was performed with the patient under conscious sedation and analgesia or general anesthesia. A single lesion was treated in 16 patients, and multiple lesions were treated in eight patients. Follow-up with serial computed tomography ranged from 4 to 44 months (mean, 10 months; median, 19 months). RESULTS: Complete necrosis was achieved in 59 (92%) of 64 lesions. Among the 59 lesions, complete necrosis required a single treatment session in 58 lesions (92%) and two treatment sessions in one lesion (2%). In 14 (58%) of 24 patients, new metastases developed during follow-up. Ten (71%) of these 14 patients developed new liver metastases. Currently, 10 (63%) of 16 patients whose lesions were initially confined to the liver are free of disease. One patient died of progressive brain metastases. No major complications occurred. Two minor complications were observed. CONCLUSION: On the basis of preliminary study results, percutaneous RF ablation appears to be a simple, safe, and effective treatment for focal liver metastases in selected patients with breast cancer. PMID- 11425988 TI - Portal venous stent placement in patients with pancreatic and biliary neoplasms invading portal veins and causing portal hypertension: initial experience. AB - PURPOSE: To evaluate the clinical usefulness of portal venous stent placement in patients with pancreatic or biliary neoplasms invading portal veins and causing portal hypertension. MATERIALS AND METHODS: Thirteen patients underwent portal venous stent placement because of gastrointestinal bleeding (n = 8), risk of gastroesophageal varix rupture (n = 4), ascites (n = 4), thrombocytopenia (n = 3), and/or portal venous thrombosis (n = 3). The main portal vein or both the intrahepatic and main portal veins were invaded in six patients (group A). The main portal vein and splanchnic veins were involved in seven patients (group B). Stents were placed across the stenotic (n = 8) or occluded (n = 5) lesions after percutaneous transhepatic portography. Changes in portal venous pressure, stent patency, and survival were evaluated. RESULTS: Mean portal venous pressure decreased significantly immediately after stent placement, from 24.9 mm Hg +/- 5.9 (SD) to 15.8 mm Hg +/- 4.6 (P <.001). In group A, blood flow through the stent was maintained and the symptoms had subsided at follow-up (mean, 12.5 months). In group B, symptoms were improved in five patients, but the stents were occluded in all but one patient at a mean follow-up of 1.5 months. There was a significant difference in stent patency between the patients with (14%) and those without (100%) splanchnic venous involvement (P <.01). CONCLUSION: Stent placement helped to relieve portal hypertension symptoms. Splanchnic venous involvement was associated with worse stent patency. PMID- 11425989 TI - Analysis of initial CT findings after endovascular repair of abdominal aortic aneurysm. AB - PURPOSE: To determine the spectrum and frequency of specific computed tomographic (CT) findings in the acute period after endovascular repair of abdominal aortic aneurysm (AAA). MATERIALS AND METHODS: CT images obtained 1--3 days after endograft placement were evaluated in 88 patients. The images were analyzed for stent position, appearance of endograft components, perigraft leak, and postoperative findings including air and acute thrombus within the aneurysm and air surrounding the femoral-femoral bypass graft. Findings that could be misinterpreted as perigraft leak were evaluated. RESULTS: Fifteen (17%) of 88 patients had perigraft leak in the acute postoperative period. The bare segment of the proximal self-expanding stent covered one or both renal arteries in 54 (61%) patients. One patient had CT evidence of renovascular compromise. Postoperative air was within the aneurysmal sac in 51 (58%) patients and surrounded the femoral-femoral bypass graft in 67 (94%) of 71 patients in whom the grafts were evaluated with CT. Mottled attenuation within the aneurysmal sac was seen in 50 (57%) patients. Forty-six (52%) patients had calcifications within longstanding thrombus. In 31 (35%) patients, findings that could have been misinterpreted as perigraft leak were identified. CONCLUSION: Accurate analysis of CT findings after endovascular AAA repair requires careful review of all available CT images (preprocedural and pre- and postcontrast) and clear understanding of specific stent-graft components and placement. PMID- 11425990 TI - CT fluoroscopy--guided interventional procedures: techniques and radiation dose to radiologists. AB - PURPOSE: To determine the radiation dose to radiologists who perform computed tomographic (CT) fluoroscopic interventional procedures by using a quick-check method and a low-milliampere technique. MATERIALS AND METHODS: Two hundred twenty CT fluoroscopy--guided interventional procedures were performed in 189 patients. Procedures included 57 spinal injections, 17 spinal biopsies, 24 chest biopsies, 20 abdominal aspirations, 44 abdominal biopsies, and 58 abdominal drainages. Procedure details were prospectively recorded and included site, depth, target diameter, milliampere value, kilovolt peak, fluoroscopic time, and CT technique (continuous CT fluoroscopy, quick-check method, or a combination of these techniques). An individual collar and finger radiation detector were worn by each radiologist during each procedure to determine the dose per procedure. RESULTS: The quick-check technique was performed in 191 (87%) of 220 procedures. Four procedures were performed with continuous CT fluoroscopy, and a combination technique was used for 25 (11%) procedures. The overall mean CT fluoroscopic time was 17.9 seconds (range, 1.2--101.5 seconds). The mean milliampere value was 13.2 mA (range, 10--50 mA). The overall mean radiologist radiation dose per procedure was 2.5 mrem (0.025 mSv) (whole body). Individual procedure doses ranged from 0.66 to 4.75 mrem (0.007--0.048 mSv). The finger radiation dose was negligible. CONCLUSION: By using a low-milliampere technique and the quick-check method, CT fluoroscopic time and radiation exposure can be minimized. PMID- 11425991 TI - Renal artery stenosis: duplex US after angioplasty and stent placement. AB - PURPOSE: To evaluate the hemodynamic outcome of technically successful percutaneous transluminal renal artery angioplasty and stent placement (PTRAS) with duplex ultrasonography (US). MATERIALS AND METHODS: Eighteen patients who underwent PTRAS in 22 renal arteries were prospectively examined. All had abnormal preprocedural duplex US findings. Those who had significant renal artery stenosis (>70%) at angiography and underwent technically successful percutaneous interventions were enrolled. Standard intrarenal duplex US parameters (acceleration index [AI], acceleration time, waveform morphology grade, and resistive index) were compared before and after interventions. RESULTS: A significant AI increase occurred after PTRAS (9.02 m/sec(2) +/- 4.85 [SD]), as compared with before intervention (2.34 m/sec(2) +/- 2.03; P <.001). Acceleration time significantly decreased from 0.084 second +/- 0.049 to 0.032 second +/- 0.008 (P <.01). There was also a significant resistive index increase from 0.69 +/- 0.12 to 0.79 +/- 0.12 (P <.01). Abnormal waveform morphology (modified Halpern waveform grade 3-6) was present in 19 (86%) of 22 intrarenal arteries prior to intervention, as compared with one (5%) after PTRAS (P <.001). In the instance in which an abnormal waveform persisted after intervention, waveform morphology improved from grade 6 to grade 3, with a concomitant AI increase from 0.96 to 5.1 m/sec(2). CONCLUSION: The findings suggest an important potential role for duplex US in noninvasive assessment of the immediate hemodynamic outcome and long-term follow-up of PTRAS. PMID- 11425992 TI - Nonstenotic internal carotid arteries: effects of age and blood pressure at the time of scanning on Doppler US velocity measurements. AB - PURPOSE: To assess the effects of age and blood pressure at the time of scanning on internal carotid artery velocities and cross-sectional diameter at Doppler ultrasonography (US). MATERIALS AND METHODS: During 12 months, 1,020 consecutive patients underwent internal carotid artery Doppler US. No or minimal arterial disease was found in 142 patients (67 women, 75 men). Blood pressure was recorded prior to examination. The angle-corrected internal carotid artery peak systolic and end-diastolic velocities were obtained. The effects of systolic blood pressure, diastolic blood pressure, pulse pressure, age, chronic hypertension, and medications for hypertension on velocities were evaluated by using linear regression analysis. RESULTS: Peak systolic velocity was influenced by age (P =.008), systolic blood pressure (P =.009), diastolic blood pressure (P =.003), and pulse pressure (P =.017) but not history of hypertension (P =.53) or antihypertensive medication use (P =.77). Increasing age decreased peak systolic velocity by 0.34 cm/sec/y. End-diastolic velocity was influenced by age (P <.001) but not by systolic, diastolic, or pulse pressure (all P values were >.13). CONCLUSION: Internal carotid artery peak systolic velocities decrease with advancing age and increase with increasing pulse pressure. The effects of blood pressure at the time of scanning are small, but isolated systolic hypertension could cause increases in spurious velocity. PMID- 11425993 TI - Carotid artery stenosis: prospective comparison of CT, three-dimensional gadolinium-enhanced MR, and conventional angiography. AB - PURPOSE: To prospectively compare gadolinium-enhanced magnetic resonance (MR) angiography and computed tomographic (CT) angiography with digital subtraction angiography (DSA) for use in detecting atheromatous stenosis and plaque morphology at the carotid bifurcation. MATERIALS AND METHODS: Forty-four carotid arteries (in 22 patients) were analyzed by using CT angiography, enhanced MR angiography, and DSA. CT and enhanced MR angiograms were reconstructed with maximum intensity projection and multiplanar volume reconstruction. The following four features were analyzed: degree of stenosis on the basis of North American Symptomatic Carotid Endarterectomy Trial criteria, length of stenosis, luminal surface, and presence of ulcers. RESULTS: There was significant correlation between CT angiography, enhanced MR angiography, and DSA for degree and length of stenosis. With enhanced MR angiography and CT angiography, degree of stenosis was underestimated in two of 44 cases. No case of overestimation with CT angiography was found. Severe internal carotid artery stenoses were detected with high sensitivity and specificity: 100% and 100%, respectively, with CT angiography; 93% and 100%, respectively, with enhanced MR angiography. Luminal surface irregularities were most frequently seen at CT angiography. With CT angiography and enhanced MR angiography, more ulceration was detected than with DSA. CONCLUSION: There was a significant correlation between CT angiography, enhanced MR angiography, and DSA in evaluation of carotid artery stenosis. Enhanced MR angiography or CT angiography can be used to adequately evaluate carotid stenosis. PMID- 11425994 TI - Lingual vascular canals of the mandible: evaluation with dental CT. AB - PURPOSE: To evaluate whether dental computed tomography (CT) can depict the frequency, diameter, position, and direction of vessels entering the mandible through lingual bone canals. MATERIALS AND METHODS: Thirty-two consecutive patients underwent preimplantation CT of the lower jaw and examination for the presence, number, location, diameter, and direction of lingual canals entering the mandible. In addition, three cadaver mandibles were investigated with dental CT and subsequently dissected to confirm the CT findings. RESULTS: All patients demonstrated at least one lingual vascular canal, and 20 (63%) had multiple (two to five) canals. The typical lingual canal locations were the midline of the mandible and the premolar region. The mean diameter of the lingual canals was 0.7 mm +/- 0.3 (SD) (range, 0.4--1.5 mm) in the midline and 0.6 mm +/- 0.2 (range, 0.3--1.2 mm) in both premolar regions of the mandible. Examination results in the three cadaver mandibles confirmed the CT findings in those mandibles. CONCLUSION: Dental CT can depict the occurrence, position, and size of the lingual vascular canals of the mandible. Radiologists should be aware of this anatomic feature and its possible implications. PMID- 11425996 TI - Case 36: bilateral cervical spondylolysis of C6. PMID- 11425997 TI - Severe hemodynamic impairment and border zone--region infarction. AB - PURPOSE: To investigate the relationship between the patterns of cerebral infarction that have been associated with hemodynamic impairment and the presence of severe chronic hemodynamic compromise (increased oxygen extraction fraction) in a large prospectively enrolled group of patients with carotid artery occlusion. MATERIALS AND METHODS: At enrollment in a prospective study of cerebral hemodynamics, 110 patients with carotid occlusion underwent (a) positron emission tomography for the measurement of cerebral oxygen extraction fraction and (b) computed tomographic (CT) or magnetic resonance (MR) examinations of the brain. Infarcts were categorized retrospectively by vascular territory, location, and pattern. The association of these findings with hemodynamic impairment (increased oxygen extraction fraction) was investigated. RESULTS: No border zone region infarctions were found in 35 asymptomatic patients. In 75 symptomatic patients, cortical border zone-region infarction was found in seven of 36 patients with increased oxygen extraction fraction, and in two of 39 with normal oxygen extraction fraction (P =.08, difference not significant). The pattern of multiple white matter lesions arranged parallel to the lateral ventricle was observed only in symptomatic patients with increased oxygen extraction fraction (eight of 36 patients; P =.002; sensitivity, 22%; specificity, 100%). This finding was more frequent with MR imaging (seven of 14 patients) than with CT (one of 22 patients). CONCLUSION: Multiple white matter infarctions, arranged parallel to the lateral ventricle, are associated with severe hemodynamic impairment. This pattern of infarction is likely due to a hemodynamic mechanism. PMID- 11425998 TI - Temporal lobe activation demonstrates sex-based differences during passive listening. AB - PURPOSE: To evaluate potential sex differences in temporal lobe activation during the performance of a functional magnetic resonance (MR) imaging passive-listening paradigm. MATERIALS AND METHODS: Twenty strongly right-handed volunteers (10 men, 10 women) underwent imaging with a 1.5-T machine by using a gradient-echo echo planar sequence. The task consisted of passive listening to simple narrative text interleaved with same-narrative text played backward. Volumes of interest were drawn around anterior and posterior areas of activation in bilateral temporal lobes. The peak percentage of activation and the percentage of activated voxels at single-voxel significance levels of 10(-2), 10(-3), and 10(-4) within each volume of interest were measured. An asymmetry index A was then calculated for both anterior and posterior volumes of interest such that A = (L - R)/(L + R), where R is either the peak percentage activation or the percentage of activated voxels within the right volume of interest and L is either the peak percentage activation or the percentage of activated voxels within the left volume of interest. The asymmetry indexes were compared between men and women by using a standard t test. RESULTS: Men showed a significantly higher degree of asymmetric activation than did women in both the anterior and posterior volumes of interest by using peak percentage activation and at all single-voxel significance levels. The degree of activation asymmetry was greater by using single-voxel significance measurements, compared with peak percentage activation measures. CONCLUSION: Women demonstrate a higher degree of bilateral language representation in temporal lobe regions than do men during passive listening. These findings, combined with the variable results of prior functional MR imaging language studies of sex differences, suggest that they may be task specific. PMID- 11425999 TI - Medullary cone movement in subjects with a normal spinal cord and in patients with a tethered spinal cord. AB - PURPOSE: To compare movement of the normal medullary cone when the patient has changed from a supine to prone position with that in patients with known or suspected tethered spinal cord syndrome. MATERIALS AND METHODS: Fifty-six individuals divided into three groups were examined with lumbar spine magnetic resonance (MR) imaging performed with the patient in the prone and supine positions. Group 1 consisted of 15 healthy volunteers and six patients with a herniated disk; group 2, 25 patients clinically suspected of having a tethered cord; and group 3, 10 patients who previously had undergone tethered cord surgery. RESULTS: All group 1 subjects showed distinct and statistically significant medullary cone movement (range, 21%--41%); no patient in group 3 showed movement (Wilcoxon rank sum test, P <.001). In group 2, the 20 patients in whom a definite diagnosis of tethered cord syndrome was made on the basis of initial supine MR image findings showed no movement, whereas two of five patients with normal supine MR images had abnormal and decreased cone movement at prone imaging. CONCLUSION: Prone MR imaging has no additional value when the supine MR image has clearly shown the cause of tethering or in patients who have undergone tethered cord surgery, but it can provide additional information in patients clinically suspected of having a tethered cord and in whom supine MR imaging depicted no abnormalities. PMID- 11426000 TI - Vertebral bone marrow perfusion evaluated with dynamic contrast-enhanced MR imaging: significance of aging and sex. AB - PURPOSE: To investigate blood perfusion of nonfractured, normal-appearing vertebral bodies with regard to age and sex. MATERIALS AND METHODS: Dynamic magnetic resonance imaging (160 images obtained in 80 seconds) was performed from T10 to L5 in 66 patients. Patients were assigned to three groups: group 1, those 50 years or younger without compression fracture; group 2, those older than 50 years without compression fracture; or group 3, those older than 50 years with compression fracture. Peak enhancement percentage and enhancement slope were determined from the time-intensity curve of normal (nonfractured) vertebral body. Comparisons were made between groups, and the effect of age and sex interaction was analyzed. RESULTS: Higher peak enhancement percentage was demonstrated for group 1 compared with group 2 (58.21 +/- 44.65 [SD] vs 21.88 +/- 14.77, P <.005). Group 1 women revealed a higher enhancement percentage compared with group 1 men (87.17 +/- 54.13 vs 38.16 +/- 21.69, P <.05), which significantly decreased in those older than 50 years (from 87.17 +/- 54.13 to 17.98 +/- 13.80, P <.005). For men, this decrease in those older than 50 years was not as pronounced (from 38.16 +/- 21.69 to 25.38 +/- 15.43, P >.05). Presence of compression fracture at other levels of the spine (group 3) was not associated with a different enhancement percentage for normal vertebrae. CONCLUSION: Rate of vertebral bone marrow perfusion revealed a significant decrease in subjects older than 50 years. Women demonstrated a higher marrow perfusion rate than men younger than 50 years and a more marked decrease than men older than 50 years. PMID- 11426001 TI - MR arthrography of the glenohumeral joint: two concentrations of gadoteridol versus Ringer solution as the intraarticular contrast material. AB - PURPOSE: To compare two concentrations of gadoteridol with Ringer solution as the contrast material for magnetic resonance (MR) arthrography of the glenohumeral joint. MATERIALS AND METHODS: One hundred fifty-six consecutive MR arthrograms were randomly obtained with either 2 mmol/L gadoteridol (n = 52), 4 mmol/L gadoteridol (n = 52), or Ringer solution (n = 52). MR arthrograms were assessed quantitatively (for contrast-to-noise ratio [CNR]) and qualitatively (for overall image quality, image contrast, degree of joint distention, and motion artifacts). MR diagnoses were compared with arthroscopic or surgical reports in 88 patients. RESULTS: The mean CNR at imaging was 40.4 with 2 mmol/L gadoteridol, 45.6 with 4 mmol/L gadoteridol, and 48.7 with Ringer solution. The CNR with 2 mmol/L gadoteridol was significantly lower than that with 4 mmol/L gadoteridol (P =.025) and Ringer solution (P =.012). Qualitative differences between the two gadoteridol concentrations were not significant. Ringer solution was significantly worse with regard to overall quality, motion artifacts, image contrast, and joint distention compared with both gadoteridol concentrations. Ringer solution was slightly more sensitive and less specific than the gadoteridol solutions in the detection of supraspinatus tears and less sensitive and more specific in enabling diagnosis of superior labrum anteroposterior lesions. CONCLUSION: MR arthrograms of the shoulder obtained with gadoteridol and those obtained with Ringer solution provided equivalent diagnostic accuracy. The authors, however, preferred the image quality of the gadoteridol-enhanced arthrograms. PMID- 11426002 TI - Intramedullary osteosclerosis: imaging features in nine patients. AB - PURPOSE: To determine the conventional radiographic, computed tomographic (CT), magnetic resonance (MR) imaging, scintigraphic, and histologic features of intramedullary osteosclerosis and to review the clinical features. MATERIALS AND METHODS: Nine female patients with leg pain and imaging features indicative of intramedullary sclerosis were seen during a 25-year period. None of the patients had a history of trauma or infection, familial bone disease, or related abnormal laboratory findings. Imaging studies included radiography (n = 9), CT (n = 4), MR imaging (n = 5), and skeletal scintigraphy (n = 5). Histologic correlation was available in five patients. RESULTS: Sixteen bone lesions (midtibia, n = 14; distal fibula, n = 1; and proximal femur, n = 1) were evident. Both lower extremities were involved in seven patients, and a single extremity was involved in two. Intramedullary sclerosis was present, as was cortical thickening, mainly in the diaphysis of the long bones, without extensive periosteal reaction or soft tissue involvement. Findings at bone scintigraphy were positive in all lesions. Histologic analysis showed nonspecific changes of markedly sclerotic bone with a variable degree of mineralization and maturity. CONCLUSION: Intramedullary osteosclerosis is a distinct disorder that typically affects the diaphysis of one or both tibiae in women. Characteristic imaging findings, when coupled with clinical information, allow precise diagnosis. PMID- 11426003 TI - Scaphoid fractures: evaluation with high-spatial-resolution US initial results. AB - PURPOSE: To evaluate the diagnostic accuracy of high-spatial-resolution ultrasonography (US) in the diagnosis of scaphoid fractures. MATERIALS AND METHODS: In 72 hours after acute wrist trauma, 15 consecutive patients were examined for possible scaphoid fractures clinically and with conventional radiographs, including scaphoid views. Thereafter, high-spatial-resolution US was performed by two experienced radiologists blinded to the results of the previously performed investigations. High-spatial-resolution US of the scaphoid bones was performed from the palmar, lateral, and dorsal directions in the longitudinal and transverse planes. US findings indicative of a scaphoid fracture were cortical discontinuity and/or periosteal elevation. Finally, magnetic resonance (MR) images (short inversion time inversion-recovery, T1- and T2* weighted) (ie, the standard) of the affected wrist were obtained and evaluated for a possible scaphoid fracture by two radiologists in consensus. RESULTS: Nine of 15 patients had scaphoid fractures. Seven (78%) of nine patients had positive findings at high-spatial-resolution US and five (56%) had such findings at conventional radiography (ie, four occult scaphoid fractures), with an accuracy of 87% and 73%, respectively. Two (50%) of four radiographically occult scaphoid fractures were depicted with high-spatial-resolution US. Sonographic findings of scaphoid fractures were either cortical discontinuity (n = 4), periosteal elevation (n = 2), or a combination of these two findings (n = 1). CONCLUSION: High-spatial-resolution US is a reliable diagnostic tool for the evaluation of occult scaphoid fractures and should be considered an adequate alternative diagnostic tool prior to computed tomography or MR imaging. PMID- 11426004 TI - Functional consequences of pleural disease evaluated with chest radiography and CT. AB - PURPOSE: To identify a system for the quantification of pleural thickening with an acceptable level of interobserver variation and good functional correlation in individuals with pleural disease. MATERIALS AND METHODS: The extent of pleural thickening and plaques was assessed in 50 patients by using the following: (a) a radiographic score based on the International Labour Office system, (b) a subjective simple computed tomographic (CT) score, (c) a subjective comprehensive CT score, (d) an objective nonautomated method, and (e) an objective computer aided semiautomated method. RESULTS: Similar correlations between the extent of diffuse pleural thickening and forced vital capacity were seen for each system (objective CT, r = -0.72, P <.001; simple CT, r = -0.69, P <.001; radiographic, r = -0.67, P <.001; comprehensive CT, r = -0.66, P <.001). Comparable correlations were observed for total lung capacity. After controlling for extent of diffuse pleural thickening, pleural plaque scores were functionally irrelevant. CONCLUSION: Comparable functional-morphologic correlations were achieved by using different CT and radiographic scoring systems for pleural disease. A subjective simple CT system had the advantages of ease of application and potential to aid in the accurate assessment of the lung parenchyma, which may be important in individuals exposed to asbestos. PMID- 11426005 TI - Intracranial arteriovenous malformations: real-time auto-triggered elliptic centric-ordered 3D gadolinium-enhanced MR angiography--initial assessment. AB - Auto-triggered elliptic centric-ordered three-dimensional (3D) gadolinium enhanced magnetic resonance (MR) angiography was compared with 3D multiple overlapping thin-slab acquisition time-of-flight (TOF) MR angiography in the evaluation of intracranial arteriovenous malformations (AVMs) in 10 patients. Intraarterial digital subtraction angiography (DSA) was the reference standard. Gadolinium-enhanced MR angiograms were found to be equivalent to DSA images in AVM component depiction in 70%--90% of cases and were consistently superior to TOF MR angiograms. PMID- 11426006 TI - Whole-body MR imaging in 30 seconds with real-time true FISP and a continuously rolling table platform: feasibility study. AB - A technique for whole-body magnetic resonance (MR) imaging in only 30 seconds was developed on the basis of a rolling table platform with integrated surface coils and real-time true fast imaging with steady-state precession. In five patients, all hepatic and pulmonary lesions with a diameter exceeding 8 mm were detected by using thoracic and abdominal helical computed tomography as the reference method. Whole-body MR imaging with real-time true fast imaging with steady-state precession is feasible and may be suitable for tumor screening and staging. PMID- 11426007 TI - Metacarpophalangeal joints in rheumatoid arthritis: laser Doppler imaging- initial experience. AB - Laser Doppler imaging is a noninvasive method yielding a spatial perfusion map. With use of a near-infrared laser, elevated perfusion associated with the metacarpophalangeal joints was detectable in patients with active rheumatoid arthritis. Findings at laser Doppler imaging correlated with pain scores and synovitis detected at ultrasonography, whereas the power Doppler sign (red pixels inside the active green box) did not. Laser Doppler imaging has the potential to help assess soft-tissue inflammation. PMID- 11426008 TI - MR imaging-guided prostate biopsy with surgical navigation software: device validation and feasibility. AB - Magnetic resonance (MR) imaging--guided prostate biopsy in a 0.5-T open imager is described, validated in phantom studies, and performed in two patients. The needles are guided by using fast gradient-recalled echo and T2-weighted fast spin echo images. Surgical navigation software provided T2-weighted images critical to targeting the peripheral zone and the tumor. MR imaging can be used to guide prostate biopsy. PMID- 11426009 TI - Characteristics of hepatic hemangiomas at contrast-enhanced harmonic US. PMID- 11426010 TI - Costs of CT. PMID- 11426011 TI - Radiology department overhead expenses. PMID- 11426014 TI - Arthritis therapy: a better time, a better day. PMID- 11426015 TI - Genetic counselling in familial Mediterranean fever: has the time come? PMID- 11426016 TI - The effects of exercise on the hormonal and immune systems in rheumatoid arthritis. PMID- 11426017 TI - Effects of the American College of Rheumatology systemic sclerosis trial guidelines on the nature of systemic sclerosis patients entering a clinical trial. AB - OBJECTIVES: To compare the systemic sclerosis (SSc) patients entered into the d penicillamine trial with SSc patients entered into previous controlled SSc trials. It was hypothesized that the d-penicillamine trial patients, who conformed to the American College of Rheumatology (ACR) guidelines for clinical trials in SSc were different from patients entered into previous trials. METHODS: Patients entering a double-blind, randomized trial of low- vs high-dose d penicillamine were described carefully and completely. Their characteristics were then compared with previously published data on SSc and its treatment. RESULTS: One hundred and thirty-four patients had early [mean duration 9.5 (s.d. 4.2) months], diffuse [skin score 21 (8)] disease. Organ involvement in the patients was as follows: pulmonary 54%, cardiac 20%, joints 38%, muscular 20%. Thirty three per cent had mild proteinuria and 13% were hypertensive when first seen. Compared with patients in most previous studies, these SSc patients had earlier disease and uniformly had diffuse disease. They had less muscular involvement, less dyspnoea, less abnormal pulmonary function and less cardiac and less renal involvement than patients in earlier studies. CONCLUSIONS: The use of the new ACR guidelines for SSc trials may change the nature of patient populations entering future studies. PMID- 11426018 TI - Osteoprotegerin and receptor activator of nuclear factor kappaB ligand (RANKL) regulate osteoclast formation by cells in the human rheumatoid arthritic joint. AB - OBJECTIVE: This study investigated the involvement of the recently identified regulators of osteoclast formation RANKL [receptor activator of nuclear factor kappaB (RANK) ligand, osteoclast differentiation factor, TRANCE, osteoprotegerin ligand] and its natural inhibitor, osteoprotegerin (OPG), in the bone erosion of rheumatoid arthritis (RA). METHODS: mRNA was extracted from cells isolated from the pannus and synovial membrane regions of joints of 11 RA patients. Semiquantitative reverse transcription-polymerase chain reaction was carried out, and the isolated cells were also cultured to determine their ability to form osteoclasts. RESULTS: mRNAs encoding RANKL, RANK, OPG and macrophage-colony stimulating factor were expressed by cells isolated from RA joints. In addition, mRNA encoding for tumour necrosis factor apoptosis-inducing ligand and the osteoclast markers tartrate-resistant acid phosphatase and calcitonin receptor were also often expressed. Osteoclasts capable of forming resorption lacunae were generated from cells in the RA joints. At 50 ng/ml, recombinant OPG completely inhibited the resorptive activity of these cells. There was a significant correlation between the ratio of RANKL mRNA to OPG mRNA and the number of resorption pits produced (P = 0.028). CONCLUSION: These data suggest that RANKL is an essential factor for osteoclast formation by cells in the rheumatic joint and that OPG may prevent the bone erosion seen in RA joints. PMID- 11426019 TI - Patient-relevant outcomes fourteen years after meniscectomy: influence of type of meniscal tear and size of resection. AB - OBJECTIVES: To study long-term patient-relevant outcomes after meniscectomy, a surgical procedure associated with a high risk of knee osteoarthritis (OA). Principal objectives were to compare traumatic with degenerative meniscal tear and partial with subtotal meniscectomy. METHODS: We studied a well-defined cohort of 205 patients who had undergone isolated unilateral meniscectomy between 1983 and 1985. There was no previous knee surgery and all knees were stable. The type of meniscal tear and surgical resection was ascertained by review of medical records. Patients were followed up after 14 yr (range 12-15 yr) by self administered questionnaires, one generic [Short Form 36 (SF-36)] and one disease specific [Knee Injury and Osteoarthritis Outcome Score (KOOS)]. RESULTS: In a multivariate analysis, using the Sports and Recreation Function and knee-related Quality of Life subscales of the KOOS questionnaire as dependent variables, patients with a degenerative tear scored significantly worse than individuals with a traumatic tear (P < or = 0.001). When we analysed unmatched subgroups and age- and sex-matched patients with degenerative or traumatic lesions, the same result was found for the knee-specific outcome (P < or = 0.02) and SF-36 except for Social Functioning (P < or = 0.04). There was no difference in outcome for the total cohort according to the type of resection. However, subgroup analyses showed that patients who underwent subtotal meniscectomy for a degenerative tear scored significantly worse on the knee-specific outcome than individuals who had had a partial meniscectomy for the same type of tear (P < or = 0.02). CONCLUSIONS: The long-term outcome of meniscal injury and surgery appears to be determined largely by the type of meniscal tear. Furthermore, our findings support the use of minimal meniscal resection in the treatment of degenerative tears. We suggest that the disease processes associated with the development of OA of the joint cartilage may also be active in the meniscus, and that a tear in a meniscus with degenerative changes might be regarded as the first sign of OA of the joint. PMID- 11426020 TI - Cardiovascular risk factors, including thrombotic variables, in a population with rheumatoid arthritis. AB - OBJECTIVE: To compare prevalent cardiovascular disease, conventional cardiovascular risk factors and thrombotic variables in a cohort with well controlled rheumatoid arthritis (RA) and in population controls. METHODS: Seventy six RA patients and 641 controls, randomly sampled from the local population in the North Glasgow MONICA study. Conventional cardiovascular risk factors (blood pressure, smoking, cholesterol) and thrombotic variables [fibrinogen, von Willebrand factor (vWF), tissue plasminogen activator antigen (t-PA), fibrin D dimer, plasminogen activator inhibitor (PAI-1), plasma viscosity] were measured by standard procedures. RESULTS: RA patients had a significantly higher prevalence of angina pectoris (P=0.03). Stroke also tended to be more common in the RA group, but the difference did not reach statistical significance (P=0.08). Diastolic blood pressure was significantly higher and serum cholesterol significantly lower in the RA group than in controls. Current smoking habits and exercise history were similar in the two groups, although RA patients were more likely to have previously smoked. Significant elevations in several thrombotic predictors of cardiovascular disease (fibrinogen, vWF, t-PA antigen and fibrin D dimer) were found in the RA group. CONCLUSIONS: In this RA patient population, diastolic blood pressure was higher than in controls and thrombotic variables were elevated compared with controls. These features are identified as potential additional cardiovascular risk factors in the RA patients studied. Prospective studies of risk modification may permit the identification of factors which could lead to a reduction in the known increased cardiovascular risk in RA. PMID- 11426021 TI - Provision of Internet-based rheumatology education (http://rheuma.bham.ac.uk). AB - OBJECTIVES: The Internet is becoming an important way of delivering medical information, and if used appropriately may assist in improving patients' self management of their disease. We have established an arthritis education website ('Arthritis Help') and investigated its use over the last 2 yr. METHODS: Computer generated log-file analysis and on-line questionnaires were used to create user profiles of our website. RESULTS: An average of 288 people visited our site each day, predominantly from America and the UK (49% of users). The typical questionnaire respondent (n = 770) was an American female with arthritis, aged 30+ yr, accessing the Internet from home. Typically, respondents had previously obtained information from medical staff or in written form, but were now more likely to use the Internet. One hundred and sixty-seven out of 585 respondents found our site to be useful, prompting them to seek more information (29%), change their behaviour or engage in more effective discussions with their physician (15%). CONCLUSIONS: These data indicate that it is possible to use the Internet to deliver medical information to its target audience, and that this process can have some impact on the way disease is self-managed. This information may aid more focused website design to maximize the use and potential benefits of such a resource. PMID- 11426022 TI - Thrombosis in Behcet's disease: a retrospective survey from a single UK centre. AB - OBJECTIVE: To contribute to knowledge of vascular occlusion in Behcet's disease (BD), prevalence and relative risk for thrombosis were ascertained retrospectively in a cohort of Caucasian patients with the disease. PATIENTS: The study included 73 BD patients (36 males, 37 females, mean age 45+/-11 yr) attending the Immunology/Rheumatology Department of Northwick Park Hospital. A group of 146 patients without BD, attending the same department, served as a control group. RESULTS: Thrombosis was more frequent in BD patients than in controls (23/73, 32% vs 7/146, 5%, P<0.001). This was accounted for by a higher prevalence of venous thrombosis in BD patients (18/73, 25% vs 4/146, 3%, P<0.001). Gender-adjusted data revealed an 11-fold risk of developing any thrombosis and a 14-fold risk of developing venous thrombosis in BD. After adjusting for differences in age at first symptoms, male BD patients showed a 6 fold higher risk of vein thrombosis. Males reported more often thrombophlebitis (13/36, 36% vs 0/0, P<0.001), folliculitis (8/36, 22% vs 1/37, 3%, P<0.01) and retinal vasculitis (13/36, 36%, vs 4/37, 11%, P=0.01) than females, in whom arthralgia prevailed (23/37, 62%, vs 12/36, 33%, P=0.01). CONCLUSION: In our population, BD confers a 14-fold risk of developing venous thrombosis. The risk is sixfold higher in male BD patients, who fare worse than females with regard to thrombophlebitis, folliculitis and retinal vasculitis. PMID- 11426023 TI - Elevated interleukin-6 plasma levels are regulated by the promoter region polymorphism of the IL6 gene in primary Sjogren's syndrome and correlate with the clinical manifestations of the disease. AB - OBJECTIVES: To determine whether plasma interleukin-6 (IL-6) and G/C base exchange polymorphism at position -174 of the IL6 gene have an effect on the clinical manifestations of primary Sjogren's syndrome (pSS). METHODS: Levels of circulating IL-6 protein and polymorphism of the IL6 gene were analysed in 66 patients with pSS and in 400 healthy subjects. These data were studied in relation to clinical data on the pSS patients. RESULTS: Plasma IL-6 was elevated in pSS patients compared with healthy controls. pSS patients with coeliac disease, pulmonary fibrosis or alveolitis or peripheral nervous system symptoms had significantly higher IL-6 levels than patients without these manifestations. IL-6 levels increased in parallel with the histological grade of minor salivary gland biopsy and the number of pSS criteria fulfilled. IL6 allele frequencies were similar in patients and normal subjects. Plasma IL-6 levels were regulated by the IL6 genotype in pSS patients. CONCLUSIONS: The G/C polymorphism of the IL6 gene does not predispose patients to pSS, but the circulating IL-6 concentration is related to specific manifestations of the disease and the levels of IL-6 are regulated by the IL6 promoter polymorphism in pSS. PMID- 11426024 TI - Association of CTLA-4 but not CD28 gene polymorphisms with systemic lupus erythematosus in the Japanese population. AB - OBJECTIVE: Systemic lupus erythematosus (SLE) in a multisystem autoimmune disorder characterized by multiorgan pathology and autoantibodies against a variety of autoantigens. The CD28 and CTLA-4 genes might be candidate genes for SLE, because costimulation signals from CD80/CD86 to CD28/CTLA-4 have been suggested to play an important role in the activation or inactivation of T lymphocytes. METHODS: We investigated three polymorphic regions within the CTLA-4 gene, a C/T base exchange in the promoter region -318 (CTLA-4 -318C/T), an A/G substitution in the exon 1 position 49 (CTLA-4 49A/G), an (AT)(n) repeat polymorphism in the 3' untranslated region of exon 4 [CTLA-4 3' (AT)(n)], and a CD28 gene polymorphism, a T/C substitution in the intron 3 position +17 (CD28 IVS3+17T/C), in SLE patients and controls. RESULTS: SLE patients had significantly higher frequencies of the CTLA-4 49G allele (P=0.003) and of the CTLA-4 (AT)(n) 106 bp allele (P=0.0008) than controls. We also found a strong linkage disequilibrium between the A allele of CTLA-4 49A/G and the 86 bp allele of CTLA-4 3' (AT)(n). On the contrary, no association was found between SLE and CTLA-4 -318C/T or CD28 IVS3 +17T/C. CONCLUSION: We conclude that the CTLA-4 gene appears to play a significant role in the development of SLE in the Japanese population. PMID- 11426025 TI - Lack of association of HLA-B*51 with a severe disease course in Behcet's disease. AB - OBJECTIVE: To investigate the previously reported association of HLA-B51 with the manifestations and severity of Behcet's disease (BD). METHODS: The study group consisted of 148 consecutive BD patients (89 male, 59 female) with a minimum disease duration of 5 yr followed up at an out-patient BD clinic in a tertiary referral centre. The patients were classified into three severity groups (mild, moderate, severe) using a modified form of the BD total activity index. HLA-B alleles were determined by DNA amplification using the polymerase chain reaction and sequential hybridization with sequence-specific oligonucleotide probes. RESULTS: The frequencies of genital ulceration [odds ratio (OR)=3.1, 95% confidence interval (CI) 1.3-7.5], skin findings (erythema nodosum, folliculitis or acne-like lesions) (OR=4.4, 95% CI 1.1-17.7), a positive skin pathergy test (OR=3.4, 95% CI 1.1-10.9) and eye disease (OR=1.8, 95% CI 0.9-3.7) were all higher in B*51-positive patients. By contrast, no significant association was observed between B*51 positivity and a severe disease course, and B*51 homozygosity did not exhibit a prominent association with the severity of BD. Male sex was found to be the strongest determinant of the severity of BD by logistic regression analysis (OR=4.7, 95% CI 1.9-11.2). CONCLUSION: HLA-B*51 does not exhibit a strong association with a more severe disease course in BD. The involvement of other genetic and/or environmental factors seems to be required and to be more important than B*51 for the progression of BD. PMID- 11426026 TI - Osteoclast-like cells in an in vitro model of bone destruction by rheumatoid synovium. AB - OBJECTIVE: Osteoclasts may be involved in the process of rheumatoid bone destruction. To test this hypothesis, we developed an in vitro model of bone destruction by osteoclast-like cells derived from cultured rheumatoid synovial tissue without using any inducers. METHODS: Synovial tissues were obtained from rheumatoid arthritis and osteoarthritis patients and tissue pieces of about 2 mm(3) that contained synovial lining were cultured. Multinucleated cells derived from cultured synovial tissues were studied cytochemically and morphologically for osteoclast-specific markers. RESULTS: Fibroblast-like and macrophage-like cells from the tissue pieces proliferated in the coexistence of lymphocytes. After 14 days of culture, multinucleated cells with tartrate-resistant acid phosphatase activity appeared. These cells expressed vacuolar H(+)-ATPase, the vitronectin receptor and cathepsin K. Although binding of (125)I-labelled salmon calcitonin was very low, the cells contained ringed structures of F-actin and showed strong bone-resorbing activity on ivory slices. Proliferation of macrophage-like cells and formation of multinucleated cells continued during 6 months of culture in the presence of fibroblast-like cells. The bone-resorbing activity of multinucleated cells derived from rheumatoid synovial tissue was much higher than that of cells from osteoarthritis synovial tissue, and was related to the disease activity of rheumatoid arthritis. CONCLUSION: Our culture system reproduced in vitro the process of bone destruction by rheumatoid synovium, including the proliferation and fusion of precursor cells, polarization, activation and bone tissue resorption. This system may provide a tool for understanding the mechanisms of bone destruction in rheumatoid arthritis and for the development of new therapies to prevent bone destruction. PMID- 11426027 TI - Antibodies to Th/To ribonucleoprotein in patients with localized scleroderma. AB - OBJECTIVE: Antibodies to Th/To ribonucleoprotein (anti-Th/To) have been detected almost exclusively in patients with systemic sclerosis. Therefore, we aimed to determine the prevalence of anti-Th/To in patients with localized scleroderma. METHODS: Seventy serum samples from patients with localized scleroderma were examined by RNA immunoprecipitation and indirect immunofluorescence analysis using HEp-2 cells as substrate. RESULTS: Three (4%) of 70 sera from patients with localized scleroderma immunoprecipitated 7-2 (Th) RNA and 8-2 (To) RNA. Indirect immunofluorescence analysis demonstrated that all the sera positive for anti Th/To showed mainly nucleolar staining. In one patient, the coexistence of anti histone antibody with anti-Th/To was detected by enzyme-linked immunosorbent assay for anti-histone antibody and confirmed using an absorption test with histones followed by indirect immunofluorescence analysis. Moreover, the localized scleroderma patients with anti-Th/To tended to have significantly fewer sclerotic lesions than those without. CONCLUSION: Anti-Th/To is one of the serological abnormalities in localized scleroderma, and the presence of anti Th/To may be a serological indicator of a mild form of cutaneous involvement. PMID- 11426028 TI - Hyperhomocysteinaemia in Behcet's disease. AB - OBJECTIVE: Arterial and venous thrombosis are among the clinical features of Behcet's disease (BD), the pathogenesis of which is not completely understood. In this study, we investigated whether hyperhomocysteinaemia, being a well known risk factor for thrombosis, is also a contributive risk factor for the arterial and venous thrombosis of BD. METHODS: Eighty-four patients fulfilling the criteria of the International Study Group for Behcet's Disease (54 males, 30 females, mean age 36+/-9 yr) were enrolled. All the patients were carefully screened for a history of venous thrombosis and were separated into two groups with respect to thrombosis history. Thirty-six healthy individuals (23 males, 13 females), matched for age and sex with the BD group, were included as a negative control group. Patients were excluded if they had any condition that might affect plasma homocysteine concentration. As methotrexate (MTX) causes hyperhomocysteinaemia, we also included 29 rheumatoid arthritis patients (five males, 24 females) receiving MTX weekly. Fasting plasma homocysteine concentrations were measured by high-performance liquid chromatography. The data were analysed with the chi(2) test and Student's t-test. RESULTS: The highest homocysteine concentrations were found in the MTX group (17.5+/-5.3 micromol/l). Mean plasma homocysteine concentrations in BD patients were significantly higher than in the healthy controls (11.5+/-5.3 vs. 8.8+/-3.1 micromol/l, P<0.001). Among BD patients with a history of thrombosis, 20 of 31 (64%) had hyperhomocysteinaemia, and this was significantly higher than in those without thrombosis (9%). On the other hand, there was no significant difference between patients with non-thrombotic BD and healthy controls (P>0.05). In patients with thrombosis, we found no correlation between the duration of the post-thrombotic period and homocysteine concentration. Among all the variables investigated, only hyperhomocysteinaemia was found to be related to thrombosis. CONCLUSION: Hyperhomocysteinaemia may be assumed to be an independent risk factor for venous thrombosis in BD. Unlike the factor V Leiden mutation, hyperhomocysteinaemia is a correctable risk factor. This finding might lead to new avenues in the prophylaxis of thrombosis in BD. PMID- 11426029 TI - Radiographic signs of bone destruction in the arthritic temporomandibular joint with special reference to markers of disease activity. A longitudinal study. AB - OBJECTIVE: To investigate the progression of radiographic changes of the temporomandibular joint (TMJ) with reference to plasma levels of interleukin 1beta (IL-1beta), C-reactive protein (CRP) and disease duration. METHODS: Twenty one patients with chronic inflammatory joint disease and TMJ involvement were included. Individualized tomography of the TMJ was performed twice with an interval of at least 12 months. Blood samples were analysed for IL-1beta and CRP. RESULTS: Significant progression of the overall grade of radiographic changes occurred during the observation period, whereas erosions showed great interindividual variability. Progression of TMJ bone loss was correlated to raised levels of CRP and, in patients with a diagnosis of rheumatoid arthritis, or with shorter duration, also to plasma IL-1beta. CONCLUSION: Progression of overall grade of radiographic changes in the TMJ occurs in patients with chronic inflammatory joint disease. Raised levels of serum CRP are associated with progression of TMJ bone loss. PMID- 11426030 TI - Prediction of depression in systemic lupus erythematosus patients using SF-36 Mental Health scores. AB - OBJECTIVE: As depression is common in systemic lupus erythematosus (SLE) patients, we investigated whether and how the Medical Outcome Survey Short Form 36 (SF-36) scores, routinely used in the assessment of SLE patients, would indicate the absence or presence of depression. METHODS: The Depression subscale of the Hospital Anxiety and Depression Scale (HADS-D) and the SF-36 were applied in a cross-sectional cohort of 60 SLE patients [mean age 45 (S.D. 15) yr, disease duration 11 (9) yr, 90% female, 100% Caucasians]. The SF-36 domain score with the closest association with HADS-D was used for further analysis. On the basis of HADS-D scores, the patients were split into two groups: one without depression (score<8) and the other with possible depression (score > or =8). RESULTS: The SF 36 Mental Health score was most closely correlated to the depression score (rho= 0.69, P<0.0005). The calculated Mental Health score cut-off value which significantly differentiated possibly depressed from non-depressed SLE patients was 61. Its sensitivity for the detection of possible depression was 89%, its specificity 77% and its negative predictive value 97%. CONCLUSIONS: The present study contributes to knowledge of means of excluding depression and the prevention of underdiagnosis and undertreatment of depression in SLE patients. PMID- 11426031 TI - WHO Collaborating Centre consensus meeting on anti-cytokine therapy in rheumatoid arthritis. AB - Severe adult rheumatoid arthritis is a cause of progressive disability and increased mortality across Europe. A cure for the disease remains elusive, but control of symptoms and maintenance of individual independence is possible. Anti cytokine therapies offer a new approach to disease management. They are effective after the failure of full doses of methotrexate, and are at least as effective as methotrexate in retarding the progression of radiological changes. Until more is known about the long-term safety and efficacy of these drugs they should be reserved for patients with severe disease who are progressing despite adequate doses of methotrexate or other disease-modifying anti-rheumatic drugs. They should be continued until therapeutic failure or intolerance. A comprehensive health economic evaluation is needed to optimally direct the use of these drugs. This should be undertaken when long-term safety and efficacy studies are completed. PMID- 11426032 TI - Rheumatoid arthritis and cardiovascular disease may share similar risk factors. PMID- 11426033 TI - Focal myositis due to Campylobacter infection. PMID- 11426034 TI - When is a geode not a geode: when LSMFT? PMID- 11426035 TI - A severe case of acquired hypophosphataemic osteomalacia: the perils of a missed diagnosis. PMID- 11426036 TI - Acute acalculous cholecystitis and cardiac tamponade in a patient with drug induced lupus. PMID- 11426037 TI - Spinal cord compression: an unusual neurological complication of gout. PMID- 11426038 TI - Gold-induced pneumonitis: computed tomography findings in a patient with rheumatoid arthritis. PMID- 11426039 TI - Fever and dry cough in a patient with a prosthetic heart valve. An interesting presentation of temporal arteritis. PMID- 11426040 TI - Homocysteine modulation as a reason for continuous folic acid supplementation in methotrexate-treated rheumatoid arthritis patients. PMID- 11426041 TI - Occupation and upper limb disorders. PMID- 11426042 TI - Adrenal haematoma in Henoch-Schonlein purpura. PMID- 11426044 TI - Ant and human farmers face similar problems. PMID- 11426045 TI - Of fragrant shapes and colourful sounds: the world of a synaesthete. PMID- 11426046 TI - Eradication of foot-and-mouth disease: a foot in mouth proposition. PMID- 11426047 TI - Understanding intercellular communication in the brain: identified neuromuscular synapses of the fruitfly Drosophila serve as a model. PMID- 11426048 TI - Cell fate choice and social evolution in Dictyostelium discoideum: interplay of morphogens and heterogeneities. PMID- 11426049 TI - The roots of ancient medicine: an historical outline. PMID- 11426050 TI - Why genetic information processing could have a quantum basis. AB - Living organisms are not just random collections of organic molecules. There is continuous information processing going on in the apparent bouncing around of molecules of life. Optimization criteria in this information processing can be searched for using the laws of physics. Quantum dynamics can explain why living organisms have 4 nucleotide bases and 20 amino acids, as optimal solutions of the molecular assembly process. Experiments should be able to tell whether evolution indeed took advantage of quantum dynamics or not. PMID- 11426051 TI - Hydra constitutively expresses transcripts involved in vertebrate neural differentiation. AB - The diploblastic Hydra is among the most primitive multicellular organisms. Using cross-hybridization with Xenopus probes, noggin-like transcripts were detected in the hypostome and basal disc of adult Hydra (Pelmatohydra oligactis), regions with properties similar to that of the amphibian organizer. This points to the possibility of a close molecular similarity between the Xenopus and Hydra organizers. The constitutive expression of a noggin-like gene in Hydra may be responsible for its regenerative capacity. PMID- 11426052 TI - Ammonia differentially suppresses the cAMP chemotaxis of anterior-like cells and prestalk cells in Dictyostelium discoideum. AB - A drop assay for chemotaxis to cAMP confirms that both anterior-like cells (ALC) and prestalk cells (pst cells) respond to cAMP gradients. We present evidence that the chemotactic response of both ALC and pst cells is suppressed by ammonia, but a higher concentration of ammonia is required to suppress the response in pst cells. ALC show a chemotactic response to cAMP when moving on a substratum of prespore cells in isolated slug posteriors incubated under oxygen. ALC chemotaxis on a prespore cell substratum is suppressed by the same concentration of ammonia that suppresses ALC chemotaxis on the agar substratum in drop assays. Chemotaxis suppression is mediated by the unprotonated (NH3) species of ammonia. The observed suppression, by ammonia, of ALC chemotaxis to cAMP supports our earlier hypothesis that ammonia is the tip-produced suppressor of such chemotaxis. We discuss implications of ammonia sensitivity of pst cells and ALC with regard to the movement and localization of ALC and pst cells in the slug and to the roles played by ALC in fruiting body formation. In addition, we suggest that a progressive decrease in sensitivity to ammonia is an important part of the maturation of ALC into pst cells. PMID- 11426053 TI - The wings of Bombyx mori develop from larval discs exhibiting an early differentiated state: a preliminary report. AB - Lepidopteran insects present a complex organization of appendages which develop by various mechanisms. In the mulberry silkworm, Bombyx mori a pair of meso- and meta-thoracic discs located on either side in the larvae gives rise to the corresponding fore- and hind-wings of the adult. These discs do not experience massive cell rearrangements during metamorphosis and display the adult wing vein pattern. We have analysed wing development in B. mori by two approaches, viz., expression of patterning genes in larval wing discs, and regulatory capacities of larval discs following explantation or perturbation. Expression of Nubbin is seen all over the presumptive wing blade domains unlike in Drosophila, where it is confined to the hinge and the wing pouch. Excision of meso- and meta-thoracic discs during the larval stages resulted in emergence of adult moths lacking the corresponding wings without any loss of thoracic tissues suggesting independent origin of wing and thoracic primordia. The expression of wingless and distal-less along the dorsal/ventral margin in wing discs correlated well with their expression profile in adult Drosophila wings. Partially excised wing discs did not show in situ regeneration or duplication suggesting their early differentiation. The presence of adult wing vein patterns discernible in larval wing discs and the patterns of marker gene expression as well as the inability of these discs to regulate growth suggested that wing differentiation is achieved early in B. mori. The timings of morphogenetic events are different and the wing discs behave like presumptive wing buds opening out as wing blades in B. mori unlike evagination of only the pouch region as wing blades seen in Drosophila. PMID- 11426054 TI - Differential expression of syntaxin-1 and synaptophysin in the developing and adult human retina. AB - Synaptophysin and syntaxin-1 are membrane proteins that associate with synaptic vesicles and presynaptic active zones at nerve endings, respectively. The former is known to be a good marker of synaptogenesis; this aspect, however, is not clear with syntaxin-1. In this study, the expression of both proteins was examined in the developing human retina and compared with their distribution in postnatal to adult retinas, by immunohistochemistry. In the inner plexiform layer, both were expressed simultaneously at 11-12 weeks of gestation, when synaptogenesis reportedly begins in the central retina. In the outer plexiform layer, however, the immunoreactivities were prominent by 16 weeks of gestation. Their expression in both plexiform layers followed a centre-to-periphery gradient. The immunoreactivities for both proteins were found in the immature photoreceptor, amacrine and ganglion cells; however, synaptophysin was differentially localized in bipolar cells and their axons, and syntaxin was present in some horizontal cells. In postnatal-to-adult retinas, synaptophysin immunoreactivity was prominent in photo-receptor terminals lying in the outer plexiform layer; on the contrary, syntaxin-1 was present in a thin immunoreactive band in this layer. In the inner plexiform layer, however, both were homogeneously distributed. Our study suggests that (i) syntaxin-1 appears in parallel with synapse formation; (ii) synaptogenesis in the human retina might follow a centre-to-periphery gradient; (iii) syntaxin-1 is likely to be absent from ribbon synapses of the outer plexiform layer, but may occur at presynaptic terminals of photoreceptor and horizontal cells, as is apparent from its localization in these cells, which is hitherto unreported for any vertebrate retina. PMID- 11426055 TI - The Escherichia coli antiterminator protein BglG stabilizes the 5'region of the bgl mRNA. AB - The beta-glucoside utilization (bgl) genes of Escherichia coli are positively regulated by the product of the bglG gene, which functions as an antiterminator by binding to specific sequences present within the bgl mRNA. BglG is inactivated by phosphorylation in the absence of beta-glucosides by BglF, the bgl-specific component of the phosphotransferase system (PTS). Here, we present evidence for an additional function for BglG, namely the stabilization of the 5' end of the bgl mRNA. Half-life measurements of the promoter-proximal region of the bgl mRNA indicate a five fold enhancement of stability in the presence of active (unphosphorylated) BglG. This enhancement is lost when the binding of BglG to mRNA is prevented by deletion of the binding site. Interestingly, stabilization by BglG does not extend to downstream sequences. The enhanced stability of the upstream sequences suggest that BglG remains bound to its target on the mRNA even after the downstream sequences have been degraded. Implications of these observations for the mechanism of positive regulation of the operon by BglG are discussed. PMID- 11426056 TI - Functional magnetic resonance imaging of the primary motor cortex in humans: response to increased functional demands. AB - Functional magnetic resonance imaging (fMRI) studies have been performed on 20 right handed volunteers at 1.5 Tesla using echo planar imaging (EPI) protocol. Index finger tapping invoked localized activation in the primary motor area. Consistent and highly reproducible activation in the primary motor area was observed in six different sessions of a volunteer over a period of one month. Increased tapping rate resulted in increase in the blood oxygenation level dependent (BOLD) signal intensity as well as the volume/area of activation (pixels) in the contralateral primary motor area up to tapping rate of 120 taps/min (2 Hz), beyond which it saturates. Activation in supplementary motor area was also observed. The obtained results are correlated to increased functional demands. PMID- 11426057 TI - Chromium uptake by Saccharomyces cerevisiae and isolation of glucose tolerance factor from yeast biomass. AB - Fermentations with yeast Saccharomyces cerevisiae in semiaerobic and in static conditions with the addition of chromic chloride into the used molasses medium were analysed. It was proved that the addition of optimal amounts of CrCl3 into the basal medium enhanced the kinetics of alcohol fermentations. The addition of 200 mg/l CrCl3 into the medium stimulated both the yeast growth and the ethanol production in all experimental conditions. On the other hand, the results showed that Cr3+ ions were incorporated into yeast cells during fermentation. Under these conditions the accumulation of Cr3+ ions was performed by yeast cells during the exponential growth phase, and with enriched amounts of 30-45 microg/g(d.m) of cells. Yeast biomass enriched with chromium ions was extracted with 0.1 mol/l NH4OH assuming that the extracts had the glucose tolerance factor (GTF). Then the extracts were passed through a gel-filtration column in order to isolate and purify the GTF. The presence of GTF in the purified fractions was determined by measuring the absorbance at 260 nm. It is evident from the obtained results that the added purified fractions enhanced the rates of CO2 production as well as the glucose utilization during alcoholic fermentation. As expected, the enhancement of both rates depended on the amounts of extracts added to the fermentation substrate. Thus, it is evident that purified extracts contained the GTF compound, and that Cr3+ ions were bonded to the protein molecule. PMID- 11426058 TI - Dose-dependent differential effect of hemin on protein synthesis and cell proliferation in Leishmania donovani promastigotes cultured in vitro. AB - Leishmania donovani requires an exogenous source of heme for growth and transformation. In in vitro culture of the free-living promastigotes, exogenously added hemin enhances cell proliferation. In this investigation, the question of the function of heme with particular reference to protein synthesis and cell proliferation has been addressed. The results of in vitro cell culture experiments demonstrated that hemin (10 microM) alone is suitable for supporting optimum level of protein synthesis, and thereby cell proliferation of promastigotes to an extent that it can replace fetal bovine serum. However, in situ labelling experiments along with Western blots revealed that high concentration of hemin (50 microM) reduced the level of protein synthesis in general and of beta-tubulin in particular with a concomitant induction of hsp90, and induced consequent morphological changes that are observed during in situ transformation of promastigotes in mammalian macrophages. These results therefore suggest that sudden exposure to high concentration of heme in mammalian macrophages may be one of the key factors that trigger promastigote to amastigote transformation in L. donovani. Furthermore, hemin with its dual characteristic could be used as a tool to understand molecular mechanism of cell proliferation and transformation in these parasites. PMID- 11426059 TI - Olfaction in the Colorado potato beetle: ultrastructure of antennal sensilla in Leptinotarsa sp. AB - Sensillae on the antennae of the Colorado potato beetle, Leptinotarsa decemlineata are described using scanning (SEM) and transmission (TEM) electron microscopy and compared with SEM observations of antennal sensilla in L. haldemani and L. texana. In all the three species, 13 distinct sensillar types were identified with a higher density of sensilla in the more polyphagous species, L. decemlineata than in the moderately host specific L. haldemani and the highly host specific L. texana. Cuticular specializations and the predominance of olfactory sensilla are discussed in relation to host specificity in the three species. PMID- 11426060 TI - Chemistry of clitoral gland secretions of the laboratory rat: assessment of behavioural response to identified compounds. AB - The present investigations were carried out to find out the chemical nature of clitoral gland extracts and their involvement in reproductive and social behaviour. Homogenates of clitoral glands of mature estrous female rats were extracted with n-hexane and dichloromethane (1 : 1 ratio v/v) and analysed by gas chromatography linked mass spectrometry (GC-MS). Three peaks were found to be in higher concentration, which were identified as 6,11-dihydro-dibenz-b,e-oxepin-11 one (I); 2,6,10-dodecatrien-1-ol-3,7,11-trimethyl(Z) (II); and 1,2-benzene dicarboxylic acid butyl(2-ethylpropyl) ester (III). Odour preference tests demonstrated that the first compound attracted conspecifics of the opposite sex. By contrast, the second and third compounds were found to attract both sexes. The results conclude that the clitoral gland of laboratory rat contains three major chemical compounds which have a unique function in maintaining social and reproductive status. PMID- 11426061 TI - Ovarian steroid sulphate functions as priming pheromone in male Barilius bendelisis (Ham.). AB - The study reveals that pre-ovulatory females of the fish Barilius bendelisis (Ham.) release sex steroids and their conjugates into the water and that a steroid sulphate of these compounds functions as a potent sex pheromone which stimulates milt production in conspecific males prior to spawning. Since males exposed to the purified subfraction III of the steroid sulphate fraction have increased milt volume and more spermatozoa with greater motility, the function of this priming pheromone appears to be to enhance male spawning success. High turbulence and faster water currents render the hillstream ecosystem extremely challenging for chemical communication. Therefore, ovulatory female fish secrete highly water soluble steroid sulphates for rapid pheromonal action in males. Inhibited milt volume in olfactory tract lesioned (OTL) males exposed to the steroid sulphate fraction and 17alpha,20 beta-dihydroxy-4-pregnen-3-one supports the concept that the pheromonally induced priming effect in male fish is mediated through olfactory pathways. PMID- 11426062 TI - Determination of age, longevity and age at reproduction of the frog Microhyla ornata by skeletochronology. AB - Skeletochronological estimation of age, longevity, age at sexual maturity and breeding of Microhyla ornata was done. Frogs (n=62) were collected locally in August (rainy season) 1997 and brought to the laboratory. Body mass and snout vent-length (SVL) of each frog was recorded; the 4th toe of both the hind limbs was clipped under anaesthesia, fixed in 10% formalin, demineralized in 5% nitric acid and processed for histology. Limb bones (femur, humerus, tibiofibula and radioulna) of 6 large sized frogs were also processed for skeletochronology in order to study the rate of resorption. Gonads of 25 frogs (belonging to different body size ranges) were processed for histology in order to ascertain the gametogenic status of individual frogs. One to four growth rings consisting of growth zones and lines of arrested growth (LAGs) were noticed in frogs of different body sizes; the number of LAGs remained identical in all the limb bones and phalanges in 5 out of 6 frogs. Back calculation indicated that the resorption rate is very low in this frog. Male frogs possessed sperm bundles in seminiferous tubules in the 1st year, while females showed yolky follicles in the ovary in the 2nd year. Frogs found in amplexus were 3 5 years old. The results suggest that this frog may live for a maximum of 5 years in the natural population. PMID- 11426063 TI - Topoisomerase II poisoning by indazole and imidazole complexes of ruthenium. AB - Trans-imidazolium (bis imidazole) tetrachloro ruthenate (RuIm) and trans indazolium (bis indazole) tetrachloro ruthenate (RuInd) are ruthenium coordination complexes, which were first synthesized and exploited for their anticancer activity. These molecules constitute two of the few most effective anticancer ruthenium compounds. The clinical use of these compounds however was hindered due to toxic side effects on the human body. Our present study on topoisomerase II poisoning by these compounds shows that they effectively poison the activity of topoisomerase II by forming a ternary cleavage complex of DNA, drug and topoisomerase II. The thymidine incorporation assays show that the inhibition of cancer cell proliferation correlates with topoisomerase II poisoning. The present study on topoisomerase II poisoning by these two compounds opens a new avenue for renewing further research on these compounds. This is because they could be effective lead candidates for the development of more potent and less toxic ruthenium containing topoisomerase II poisons. Specificity of action on this molecular target may reduce the toxic effects of these ruthenium-containing molecules and thus improve their therapeutic index. PMID- 11426064 TI - Application of InterPro for the functional classification of the proteins of fish origin in SWISS-PROT and TrEMBL. AB - InterPro (http://www.ebi.ac.uk/interpro/) is an integrated documentation resource for protein families, domains and sites, developed initially as a means of rationalizing the complementary efforts of the PROSITE, PRINTS, Pfam and ProDom database projects. It is a useful resource that aids the functional classification of proteins. Almost 90% of the actinopterygii protein sequences from SWISS-PROT and TrEMBL can be classified using InterPro. Over 30% of the actinopterygii protein sequences currently in SWISS-PROT and TrEMBL are of mitochondrial origin, the majority of which belong to the cytochrome b/b6 family. InterPro also gives insights into the domain composition of the classified proteins and has applications in the functional classification of newly determined sequences lacking biochemical characterization, and in comparative genome analysis. A comparison of the actinopterygii protein sequences against the sequences of other eukaryotes confirms the high representation of eukaryotic protein kinase in the organisms studied. The comparisons also show that, based on InterPro families, the trans-species evolution of MHC class I and II molecules in mammals and teleost fish can be recognized. PMID- 11426065 TI - Residual HIV-1 infection during antiretroviral therapy: the challenge of viral persistence. PMID- 11426066 TI - Increased priming for interleukin-12 and tumour necrosis factor alpha in CD64 monocytes in HIV infection: modulation by cytokines and therapy. AB - BACKGROUND: A key factor leading to impaired immunity in HIV infection is an alteration of the pattern of cytokine response, although its precise nature remains controversial, particularly the in vivo influence of HIV on interleukin (IL)-12 synthesis. DESIGN: A cross-sectional study in 73 HIV-infected persons (28 of them receiving highly active antiretroviral therapy) and 18 HIV-seronegative healthy donors. METHODS: The frequency of monocytes/macrophages (M/M) synthesizing IL-12, IL-10 and tumour necrosis factor alpha (TNF-alpha) was determined in peripheral blood mononuclear cells. The cells were cultured in medium or were stimulated with lipopolysaccharide; proportions of CD64 M/M producing IL-12, TNF-alpha or IL-10 was determined by cytofluorometric analysis. The influence of exogenous interferon gamma (IFN-gamma), IL-10 or IL-15 on IL-12 synthesis was tested. RESULTS: Chronic HIV disease is associated with increased priming of M/M for IL-12 (involving both p40 and p70 molecules) and TNF-alpha synthesis; this was associated with cosynthesis of both cytokines by a fraction of M/M. Priming for IL-12 was physiologically enhanced by IFN-gamma and decreased by IL-10; IL-15 had no effect. The proportion of IL-10-producing CD64 M/M was not altered in patients compared with controls but there was an inverse correlation between IL-10-producing M/M and viral load. IL-12 production was not correlated with viral load but was increased following antiretroviral therapy. Following LPS stimulation, IL-12 and TNF-alpha responses were not altered in HIV-positive patients; however, the IL-10 response was decreased but restored by antiretroviral therapy. CONCLUSION: These observations argue for a preserved intrinsic CD64 M/M of IL-12 production in HIV pathogenesis. PMID- 11426067 TI - Dual-subtype FIV vaccine protects cats against in vivo swarms of both homologous and heterologous subtype FIV isolates. AB - OBJECTIVE: To evaluate the immunogenicity and efficacy of an inactivated dual subtype feline immunodeficiency virus (FIV) vaccine. DESIGN: Specific-pathogen free cats were immunized with dual-subtype (subtype A FIV(Pet) and subtype D FIV(Shi)) vaccine and challenged with either in vivo- or in vitro-derived FIV inocula. METHODS: Dual-subtype vaccinated, single-subtype vaccinated, and placebo immunized cats were challenged within vivo-derived heterologous subtype B FIV(Bang) [10--100 50% cat infectious doses (CID(50))], in vivo-derived homologous FIV(Shi)(50 CID(50)), and in vitro- and in vivo-derived homologous FIV(Pet)(20--50 CID(50)). Dual-subtype vaccine immunogenicity and efficacy were evaluated and compared to single-subtype strain vaccines. FIV infection was determined using virus isolation and proviral PCR of peripheral blood mononuclear cells and lymphoid tissues. RESULTS: Four out of five dual-subtype vaccinated cats were protected against low-dose FIV(Bang) (10 CID(50)) and subsequently against in vivo-derived FIV(Pet) (50 CID(50)) challenge, whereas all placebo immunized cats became infected. Furthermore, dual-subtype vaccine protected two out of five cats against high-dose FIV(Bang) challenge (100 CID(50)) which infected seven out of eight single-subtype vaccinated cats. All dual-subtype vaccinated cats were protected against in vivo-derived FIV(Pet), but only one out of five single-subtype vaccinated cats were protected against in vivo-derived FIV(Pet). Dual-subtype vaccination induced broad-spectrum virus-neutralizing antibodies and FIV-specific interferon-gamma responses along with elevated FIV specific perforin mRNA levels, suggesting an increase in cytotoxic cell activities. CONCLUSION: Dual-subtype vaccinated cats developed broad-spectrum humoral and cellular immunity which protected cats against in vivo-derived inocula of homologous and heterologous FIV subtypes. Thus, multi-subtype antigen vaccines may be an effective strategy against AIDS viruses. PMID- 11426068 TI - Lipopeptides induce cell-mediated anti-HIV immune responses in seronegative volunteers. AB - OBJECTIVE: Test the efficacy of a mixture of six NEF (N1, N2, N3), GAG (G1, G2) and ENV (E) lipopeptides in the induction of B- and T-cell anti-HIV responses. DESIGN: A randomized phase I open-label dose-finding trial. Twenty-eight healthy seronegative volunteers received the lipopeptides, with or without the adjuvant QS21. METHODS: Anti-HIV-peptide antibodies were detected by enzyme-linked immunosorbent assay and Western blotting. Induction of cellulary responses was assessed by proliferative test and (51)Cr-release assay. RESULTS: Local and systemic adverse reactions were always mild or moderate. After three injections an antibody response was detected in 25 out of 28 volunteers (89%). T cells from 19 (79%) of the 24 volunteers proliferated in response to at least one peptide. The majority of the volunteers had induced a multispecific proliferative response; that is, cells from volunteers proliferated to two (five of 19), three (five of 19), four (three of 19) or five peptides (one of 19). Cytotoxic responses by anti-HIV CD8+ lymphocytes could be tested in 24 volunteers, 13 (54%) of whom had clear and reproducible responses, with strong activity in the remaining 12 (> 20% of specific lysis), and polyepitopic responses were detected in at least seven of the 13 responders. Cytotoxic responses were found against the whole NEF protein (clade B LAI) in three of four tested volunteers and cross reactions with the proteins of clade B (MN) and clade A (Bangui) HIV-1 strains, and also HIV-2 ROD, were detected in one of two tested volunteers. CONCLUSIONS: Lipopeptides are promising immunogens for an AIDS vaccine. PMID- 11426069 TI - Cerebrospinal fluid response to structured treatment interruption after virological failure. AB - OBJECTIVE: Structured antiretroviral treatment interruption (STI) has been advocated as a therapeutic strategy for HIV-1 infection. We report initial observations of cerebrospinal fluid (CSF) HIV-1 infection in five patients undergoing serial lumbar punctures (LPs) during STI undertaken following virological failure. DESIGN AND METHODS: In this prospective observational study we quantified HIV-1 RNA concentrations and assessed both phenotypic drug susceptibility profiles and genotypic antiviral drug resistance mutations in CSF and plasma during the period of treatment interruption. CSF white blood cells were also counted, and patients' neurological status monitored. RESULTS: In four of the patients, CSF HIV-1 concentration increased more rapidly than that of the plasma, with consequent reduction in the ratio between plasma and CSF viral loads (pVL : cVL). Three individuals developed robust, though asymptomatic CSF lymphocytic pleocytosis. In all patients the predominant HIV-1 quasispecies shifted simultaneously in CSF and plasma from a drug-resistant to a more drug susceptible phenotype with identical and simultaneous changes in genotypes associated with drug resistance. CONCLUSIONS: STI may be accompanied by previously unrecognized changes in tissue viral exposures and lymphocyte traffic. Hence, despite 'virological failure' as evidenced by persistent plasma viremia, ongoing antiretroviral treatment prior to its interruption appeared to suppress CSF HIV-1 infection (indeed more effectively than that of plasma) and restrain lymphocyte traffic into the CSF. Simultaneous change of resistance mutations in CSF and plasma was likely due to re-emergence and overgrowth of pre-existing strains with ready exchange of virus between these two compartments, either facilitated by or provoking a local CSF lymphocytosis. PMID- 11426070 TI - Hepatotoxicity in HIV-1-infected patients receiving nevirapine-containing antiretroviral therapy. AB - OBJECTIVES: To assess the incidence and risk factors for hepatotoxicity associated with nevirapine. DESIGN: A prospective cohort study in a teaching and referral hospital involving all consecutive patients who were prescribed a nevirapine-containing antiretroviral regimen between September 1997 and May 2000. METHOD: Cutaneous and hepatic adverse reactions and clinical hepatitis were assessed. Blood analysis including plasma HIV-1 RNA CD4 cell counts, liver chemistry tests, and serology for hepatitis B and C viruses. Hepatotoxicity was defined as an increase of at least threefold in serum alanine aminotransferase or aspartate aminotransferase levels compared with baseline values. RESULTS: Of a total of 610 patients, 82 (13.4%) were antiretroviral naive when commencing nevirapine, and 46.2 and 8.9% were coinfected with hepatitis C and B viruses, respectively. Median duration of exposure to nevirapine was 8.7 months (interquartile range 3.4--14.3). Hepatotoxicity developed in 76 (12.5%), an incidence of 13.1/100 person-years. Kaplan--Meier estimated incidence of hepatotoxicity at 3, 6 and 12 months was 3.7, 9.7 and 20.1%, respectively. In seven (1.1%) patients, hepatotoxicity was associated with clinical hepatitis, which was reversible upon discontinuation of therapy. Multivariate analysis identified the duration of prior exposure to antiretroviral drugs, hepatitis C virus, and higher baseline levels of alanine aminotransferase as independent risk factors for hepatotoxicity. CONCLUSIONS: Hepatotoxicity but not clinical hepatitis was common in HIV-1-infected patients receiving nevirapine-containing regimens and the incidence steadily increased over time. Prolonged exposure to any antiretroviral therapy, coinfection with hepatitis C virus and abnormal baseline levels of alanine aminotransferase identified patients at a higher risk. PMID- 11426071 TI - Development of drug resistance in patients receiving combinations of zidovudine, didanosine and nevirapine. AB - OBJECTIVE: To evaluate the development of phenotypic and genotypic resistance to zidovudine, didanosine and nevirapine as a function of the virologic response to therapy in a group of drug-naive individuals receiving various combinations of these agents. DESIGN: All patients were enrolled in a double-blind controlled randomized trial (the INCAS study) and were selected for detailed resistance studies based on specimen availability and virologic response. METHODS: Within the three study groups (zidovudine/nevirapine, zidovudine/didanosine or zidovudine/nevirapine/didanosine), 16, 19 and 24 patients, respectively, had evaluable baseline isolates and remained in the study > 24 weeks. Phenotypic resistance to all three drugs was evaluated using the VIRCO recombinant virus assay. Genotypic sequencing was done on selected specimens from patients receiving zidovudine/nevirapine/didanosine. RESULTS: After 24 weeks, all available isolates taken from patients receiving nevirapine were resistant to this agent, while 18/21 (86%) patients receiving triple therapy carried such isolates at 30--60 weeks. At 24 weeks, zidovudine resistance developed in 4/40 isolates but was more frequent after 30--60 weeks, especially in patients on two drugs. The degree of zidovudine resistance (rise in concentration required for 50% inhibition) appeared lower in the triple therapy group compared with zidovudine/didanosine (P = 0.0004). All nevirapine-resistant isolates that were sequenced carried at least one mutation associated with resistance, most often K103N and/or Y181C. CONCLUSION: The use of highly active drug therapies may be associated with a beneficial effect on the development of antiretroviral drug resistance. The characteristics of virologic suppression that must be maintained to avoid resistance are currently being studied in hypothesis-driven clinical trials. PMID- 11426072 TI - Stable or increasing bone mineral density in HIV-infected patients treated with nelfinavir or indinavir. AB - BACKGROUND AND OBJECTIVES: To determine the factors contributing to changes in bone mineral density (BMD) over time in HIV-infected patients receiving highly active antiretroviral therapy (HAART). METHODS: Analyses of lumbar spine BMD in 183 male Caucasian participants in the Western Australian HIV Cohort study, comprising a longitudinal analysis of data from 54 patients on stable HAART regimens, and a cross-sectional analysis comparing data from 131 protease inhibitor (PI)-treated patients and 52 PI-naive (including 28 antiretroviral treatment-naive) patients. RESULTS: Average lumbar spine BMD remained stable or increased over the time frame considered. Although there was no evidence of a change of average BMD over time in patients receiving nelfinavir (P = 0.92), there was evidence of increasing bone density in the indinavir group (average increase, 0.31 z-score per year; P < 0.001). Lower initial z-scores in the longitudinal analysis were significantly associated with lower pre-HAART BMI (P = 0.003), consistent with results of the cross-sectional analysis in which lowest BMI prior to initial dual X-ray absorptiometry scan was associated with decreased BMD (P = 0.02, overall group). Although PI therapy was also associated with decreased BMD in a univariate analysis of the cross-sectional data (P = 0.04), this effect was abrogated in a multiple linear regression analysis (P = 0.11) with lowest BMI remaining significant (P = 0.04). CONCLUSIONS: We found no evidence, overall, of accelerated bone loss in patients treated with nelfinavir- or indinavir-containing HAART regimens, and propose that indinavir therapy may be associated with an increase in bone mineral density over time. Pre-HAART BMI was an independent and powerful determinant of an individual's initial z-score in the longitudinal analysis, and adjustment for this effect in a cross-sectional analysis abrogated the association between PI therapy and decreased lumbar spine z-score. PMID- 11426073 TI - Serotonin syndrome in HIV-infected individuals receiving antiretroviral therapy and fluoxetine. AB - OBJECTIVE: To describe HIV-infected individuals taking antidepressants who developed the serotonin syndrome due to drug--drug or drug--food interactions. DESIGN AND SETTING: Case studies carried out at the HIV Outpatient Clinic, Atlanta Veterans Affairs Medical Center. PARTICIPANTS AND INTERVENTIONS: HIV positive patients who were receiving antiretroviral and antidepressant therapies and presented with symptoms consistent with the serotonin syndrome. Their antidepressants were discontinued or the doses reduced in order to resolve the symptoms. RESULTS: Five cases of serotonin syndrome developed after patients who were taking antidepressants ingested P450 inhibitors. CONCLUSIONS: Serotonin syndrome should be suspected in patients on serotonergic medications who present with mental status change, autonomic dysfunction, and neuromuscular abnormalities. Suspicion should be heightened in those who are ingesting substances known to inhibit P450 enzymes, such as protease inhibitors, non nucleoside reverse transcriptase inhibitors, and grapefruit juice. PMID- 11426074 TI - Modelling the effect of combination antiretroviral treatments on HIV incidence. AB - OBJECTIVE: To assess the competing effects on HIV incidence in homosexual men of the decreased infectiousness of men with HIV receiving effective combination antiretroviral treatments and homosexual men engaging in unprotected anal intercourse with increased numbers of partners (levels of unsafe sex). METHODS: A mathematical model of HIV transmission in homosexual men was developed, based on the HIV epidemic in Australia in 1996, when effective antiretroviral treatments first became widely available. Uncertainties in parameters were modelled using 1000 simulations. The effect of treatments on decreasing infectiousness was randomly sampled with a median 10-fold decrease in infectiousness (range 100-fold to no decrease). Levels of unsafe sex were randomly sampled with a median 50% increase in unsafe sex (range 100% to no increase). The percentage change in HIV incidence after one year was obtained by comparison with a null model in which there was no decrease in infectiousness as a result of treatment and no change in unsafe sex. RESULTS: Results of the models suggested that whereas increased levels of unsafe sex were linearly associated with increases in HIV incidence, decreases in infectiousness because of treatments were non-linearly associated with decreases in HIV incidence. An assessment of the competing effects suggested that decreases in infectiousness of two-, five-, and 10-fold would be counterbalanced by increases in unsafe sex of approximately 40, 60 and 70%, respectively. CONCLUSION: These models suggest that apparently large decreases in infectiousness as a result of treatment could be counterbalanced in terms of new HIV infections by much more modest increases in unsafe sex. PMID- 11426075 TI - Cause-specific mortality associated with HIV and HTLV-II infections among injecting drug users in the USA. AB - BACKGROUND: Human T-lymphotropic virus type II (HTLV-II) is widespread among injecting drug users (IDU) and may contribute to the risk of leukemia/lymphoma, neurodegenerative disease, and perhaps pneumonia, especially with HIV co infection. METHODS: In 1987--1991, 6570 IDU were tested for HIV and HTLV-II antibodies. In 1998, they were matched to the National Death Index. Numbers of observed deaths of each cause were compared by standardized mortality ratios with the numbers expected, using sex-, race-, age-, and year-specific rates in the general population. Relative risk (RR) associated with each virus, compared to uninfected drug users, was estimated by Poisson modeling. RESULTS: There were 1351 deaths, including 683 (15%) of 4604 participants who enrolled seronegative for both viruses; 328 (47%) of 701 who had HIV but not HTLV-II infection; 220 (21%) of 1033 who had HTLV-II but not HIV infection; and 120 (52%) of 232 who were infected by both viruses. Compared to the general population, mortality for participants with neither virus was increased 4.3-fold [95% confidence interval (CI), 4.0--4.7] and was significantly elevated for virtually every cause of death. With HIV, mortality from medical causes, but not external causes, was increased 3.7-fold (95% CI, 3.3--4.2), particularly with AIDS and related conditions. With HTLV-II, all-cause mortality was reduced (RR, 0.8; 95% CI, 0.7- 0.9), with no statistically significant reduction or elevation for any specific cause. A non-significant excess of tuberculosis deaths (RR, 4.6; 95% CI, 0.8- 25.2) was noted with HTLV-II, but there was no excess mortality from leukemia/lymphoma, other malignancies, or neurodegenerative disease. CONCLUSIONS: Without HIV or HTLV-II, IDU had profoundly increased mortality from medical and external causes. HIV was specifically associated with death due to AIDS and related conditions. HTLV-II infection was not significantly associated with mortality from any cause, suggesting that it is not a significant human pathogen, even when present with HIV infection. PMID- 11426076 TI - Increase in the share of steady partners as a source of HIV infection: a 17-year study of seroconversion among gay men. AB - OBJECTIVES: To examine the share of steady versus casual partners as the source of HIV infection in gay male seroconversions between 1984 and 2000 and the effect of age at seroconversion on the source of HIV transmission. METHODS: The sample consisted of 144 seroconverstors from the Amsterdam Cohort Study among Homosexual Men. Questionnaires and post-seroconversion interviews were used to determine the source of HIV transmission. RESULTS: Analysis revealed an interaction effect between calendar year and age at seroconversion (P < 0.05). Younger seroconverters had higher odds ratios [odds ratio, 11.33; 95% confidence interval, 1.77--72.13] to be infected by their steady partner late in the AIDS epidemic: 15% (three of 20) between 1984 and 1987 versus 67% (six of nine) between 1994 and 2000. No such time effect was present for older seroconverters who were consistently more likely to be infected by a casual partner: 79% (37 of 47) between 1984 and 1987, and 83% (15 of 18) between 1994 and 2000. CONCLUSIONS: Young gay seroconverters today are more likely to have contracted HIV from a steady partner than from a casual partner, compared with early in the AIDS epidemic and compared also with older gay seroconverters. There is a pressing need for preventive measures addressing sexual risk behaviour within steady relationships among younger gay men. PMID- 11426077 TI - Calculating the consequences: HAART and risky sex. PMID- 11426078 TI - Viral levels in newborn African infants undergoing primary HIV-1 infection. AB - We examined weekly changes in viral levels in seven untreated infants infected with HIV at birth. Viral levels spiked immediately but reverted quickly to plateau levels typical of infant HIV infection within 2 weeks of first detected viraemia. We speculated that the depletion of naive, susceptible cells is responsible for the rapid decrease in spike levels and that the rapid replacement of lymphocytes in infants causes the high plateau viral levels (10(5) copies/ml) to be sustained. PMID- 11426079 TI - Mutations of the 3' untranslated region of the SDF1 gene in apes and monkeys: potential impact on sensitivity to AIDS induced by lentiviruses. AB - The comparison of the stromal cell-derived factor-1 (SDF1) gene 3' untranslated region (3'UTR) of four great ape and four monkey species with their human counterparts shows that the human SDF1-3'A mutation is present in primate species that are the most susceptible to lentivirus-induced AIDS and is absent in species that are particularly resistant to lentivirus-induced AIDS. The results enlighten the possible relationship between SDF1-3'UTR polymorphism and sensitivity to AIDS. PMID- 11426080 TI - No evidence that vaccination with a polysaccharide pneumococcal vaccine protects drug users against all-cause pneumonia. PMID- 11426081 TI - Discordant CD4 T lymphocyte responses to antiretroviral therapy for HIV infection are associated with ex-vivo rates of apoptosis. AB - Our purpose was to determine if changes in CD4 cell counts in HIV-infected patients with good viral suppression on stable antiretroviral regimens could be predicted by ex-vivo rates of apoptosis of peripheral blood mononuclear cells (PBMC). Patients were grouped by lowest pre-treatment and highest on-treatment CD4 cell counts and classified as complete immune responders, partial responders, or non-responders. Whole blood was collected from a subgroup of patients and controls, and rates of the ex-vivo apoptosis of PBMC were assessed. Non responders exhibited significantly increased apoptosis, whereas good immune responses were associated with decreased apoptosis. Persistently accelerated apoptosis may contribute to persisting immune deficiency independent of the viral load. PMID- 11426082 TI - Time trend in incidence of HIV seroconversion among homosexual men repeatedly tested in Madrid, 1988--2000. AB - An open cohort of 2670 homosexual men repeatedly tested for HIV at a Madrid clinic has registered 8050 person-years (PY) of follow-up and 157 seroconversions from 1988 to 2000. After declining from 1988 (4.71 per 100 PY), the incidence rate began a significant upward trend, reaching a figure of 2.16 per 100 PY in 2000. These findings ought to alert surveillance systems and prevention programmes. PMID- 11426083 TI - Increasing incidence of HIV infections among young gay and bisexual men in Vancouver. AB - Since the beginning of the HIV epidemic in north America, the majority of HIV infections have occurred among men who engage in sexual relations with other men. As the HIV epidemic enters its third decade, gay and bisexual men continue to have among the highest rates of HIV infection. Previous studies have highlighted the decline in the incidence of HIV and risk behaviour among gay and bisexual men. However, several studies have suggested that young gay and bisexual men continue to engage in unprotected sexual behaviours and are at continued risk of HIV infection. Recent reports in the media and research literature have indicated an increase in the incidence of HIV among gay and bisexual individuals in many of the world's major cities. The purpose of this study was to determine trends in HIV incidence using data from a prospective cohort of young gay and bisexual men. PMID- 11426084 TI - Management of sudden psychiatric disorders related to efavirenz. PMID- 11426085 TI - The abacavir hypersensitivity reaction and interruptions in therapy. PMID- 11426086 TI - Reply to 'Methods of feeding and transmission of HIV-1 from mothers to children by 15 months of age: prospective cohort study from Durban, South Africa' by A. Coutsoudis, K. Pillay, L. Kuhn, E. Spooner, W.-Y. Tsai, H. Coovadia, for the South African Vitamin A Study Group. PMID- 11426088 TI - Mother-to-child transmission of HIV-1 and infant feeding practice. PMID- 11426089 TI - Notes and quotes. New drugs give hint of promise in treating HIV/AIDS. PMID- 11426090 TI - A case in point. PMID- 11426091 TI - Effect of a subanesthetic dose of intravenous ketamine and/or local anesthetic infiltration on hemodynamic responses to skull-pin placement: a prospective, placebo-controlled, randomized, double-blind study. AB - Insertion of cranial pins for stabilization of the head can result in a marked hypertensive response, which may adversely affect cerebral hemodynamics. The efficacy of a subanesthetic dose of intravenous ketamine (0.5 mg/kg) and/or lidocaine infiltration (1%) at pin fixation sites before pinning was studied in a prospective, double-blind, placebo-controlled, randomized trial of 40 patients. The subjects were divided into four groups of 10. Patients belonging to the placebo and lidocaine groups received intravenous normal saline (NS), followed by local infiltration with NS at pin insertion sites in the placebo group and 1% lidocaine in the lidocaine group. Patients belonging to the ketamine and ketamine lidocaine groups received intravenous ketamine followed by local infiltration with NS in the ketamine group, and lidocaine infiltration in the ketamine lidocaine group. Heart rate (HR) and invasive mean blood pressure (MBP) were recorded before intravenous medication and then at various time intervals until 15 minutes after pin fixation. Intergroup comparison of MBP and HR by 2-way analysis of variance revealed a significant difference between the groups and various time points (P < .05). Post hoc analysis revealed maximum increase in MBP each hour in the placebo group. Mean blood pressure response in the ketamine group was similar to the placebo group. Significant attenuation of MBP and HR was observed in the lidocaine and ketamine-lidocaine groups (P < .05). A minimal increase in HR was observed in the lidocaine-ketamine group. The current study demonstrates maximum attenuation of hemodynamic responses when a subanesthetic dose of intravenous ketamine (0.5 mg/kg) is administered with 1% lidocaine infiltration. PMID- 11426092 TI - Effects of dihydroergotamine on intracranial pressure, cerebral blood flow, and cerebral metabolism in patients undergoing craniotomy for brain tumors. AB - In a search for a nonsurgical intervention to control intracranial hypertension during craniotomy, the authors studied the effects of dihydroergotamine on mean arterial blood pressure (MABP), intracranial pressure (ICP), cerebral perfusion pressure (CPP), cerebral blood flow (CBF), and cerebral metabolism in patients who underwent craniotomy for supratentorial brain tumors. Twenty patients were randomized to receive either dihydroergotamine 0.25 mg intravenously or placebo as a bolus dose during craniotomy. Anesthesia was induced with thiopental/fentanyl/atracurium, and maintained with isoflurane/N2O/fentanyl at normocapnia. After removal of the bone flap and exposure of intact dura, ICP was measured subdurally and dihydroergotamine/placebo was administered. Intracranial pressure and MABP were measured continuously. Cerebral blood flow (after intravenous administration of 133Xe) and arteriojugular venous difference of oxygen (AVDO2) were measured before, and 30 minutes after, dihydroergotamine/placebo administration. Cerebral metabolic rate of oxygen (CMRO2) was calculated. After administration of dihydroergotamine, a significant increase in MABP from 74 to 87 mm Hg (median) and CPP from 65 to 72 mm Hg (median) were found. Simultaneously to the increase in MABP, a significant increase in ICP from 9.5 to 11.5 mm Hg (median) was disclosed, whereas no significant differences in CBF, AVDO2, or CMRO2 were found. Intracranial pressure was significantly higher after dihydroergotamine than after placebo. In conclusion, no ICP decreasing effect of a bolus dose of dihydroergotamine was found when administered to patients with brain tumors during isoflurane/N2O anesthesia. Corresponding increases in MABP and ICP suggest that abolished cerebral autoregulation might explain why dihydroergotamine was associated with an ICP increase. PMID- 11426093 TI - Cerebral blood flow velocity response to magnesium sulfate in patients after subarachnoid hemorrhage. AB - Magnesium sulfate therapy, standard in preventing seizures in preeclampsia, is under active investigation as a neuroprotective agent. The authors studied the effect of magnesium as a cerebral vasodilator by measuring the cerebral blood flow velocity (CBFV) response to a 5g intravenous bolus of MgSO4 compared with a saline placebo after subarachnoid hemorrhage (SAH). Transcranial Doppler ultrasonography of the middle cerebral artery (MCA) was measured after each infusion. Patients were studied up to three times after SAH at prescribed time intervals. Fourteen patients (11 women, 3 men; mean age 58 years) underwent 29 studies. All patients underwent hypertensive, hypervolemic therapy. Four patients developed cerebral vasospasm. Doubling serum magnesium levels did not affect MCA CBFV but slightly lowered mean arterial blood pressure and systemic vascular resistance. Intravenous magnesium bolus did not reduce elevated CBFV in the subset of SAH patients with clinical vasospasm. The role of magnesium sulfate as a cerebral vasodilator in patients with SAH requires further study. PMID- 11426094 TI - Effect of prophylactic ondansetron on postoperative nausea and vomiting after elective craniotomy. AB - This prospective, randomized, placebo-controlled, double-blind study was designed to evaluate the efficacy of ondansetron, a 5-HT3 antagonist, in preventing postoperative nausea and vomiting (PONV) after elective craniotomy in adult patients. The authors also tried to discover certain predictors for postcraniotomy nausea and vomiting. We studied 170 ASA physical status I and II patients, aged 15 to 70 years, undergoing elective craniotomy for resecting various intracranial tumors and vascular lesions. A standardized anesthesia technique and postoperative analgesia were used for all patients. Patients were divided into two groups and received either saline placebo (Group 1) or ondansetron 4 mg (Group 2) intravenously at the time of dural closure. Patients were extubated at the end of surgery and episodes of nausea and vomiting were noted for 24 hours postoperatively in the neurosurgical intensive care unit. Demographic data, duration of surgery, and anesthesia and analgesic requirements were comparable in both groups. Overall, a 24-hour incidence of postoperative emesis was significantly reduced in patients who received ondansetron compared with those who received a saline placebo (39% in Group 1 and 11% in Group 2, P = .001). There was a significant reduction in the frequency of emetic episodes and rescue antiemetic requirement in patients treated with ondansetron; however, ondansetron did not significantly reduce the incidence of nausea alone (14% in Group 2 vs 5% in Group 1, P = .065). Prophylactic ondansetron had a favorable influence on PONV outcome measures such as patient satisfaction and number needed to prevent emesis (3.5). Side effects were similar in both groups. We conclude that ondansetron 4 mg given at the time of dural closure is safe and effective in preventing emetic episodes after elective craniotomy in adult patients. PMID- 11426095 TI - Effects of alterations in arterial CO2 tension on cerebral blood flow during acute intracranial hypertension in rats. AB - Cerebrovascular reactivity to CO2 in clinical and experimental studies has been found to be impaired during increased intracranial pressure (ICP). However, from previous study results it has not been possible to estimate whether the impairment was caused by elevated ICP, or caused by decreased cerebral perfusion pressure (CPP). The current study was carried out in a group of unmanipulated control rats and in six investigation groups of six rats each: two groups with elevated ICP (30 and 50 mm Hg) and spontaneous arterial blood pressure (MABP), two groups with spontaneous ICP and arterial hypotension (77 and 64 mm Hg), and two groups with elevated ICP (30 and 50 mm Hg) and arterial hypertension (124 mm Hg). Intracranial hypertension was induced by continuous infusion of lactated Ringer's solution into the cisterna magna, arterial hypotension by controlled bleeding, and arterial hypertension by continuous administration of norepinephrine intravenously. Cerebral blood flow (CBF) was measured repetitively by the intraarterial 133Xe method at different levels of arterial PCO2. In each individual animal, CO2 reactivity was calculated from an exponential regression line obtained from the corresponding CBF/PaCO2 values. By plotting each individual value of CO2 reactivity against the corresponding CPP value from the seven investigation groups, CPP was significantly and directly related to CO2 reactivity of CBF (P < .001). No correlation was found by plotting CO2 reactivity values against the corresponding MABP values or the corresponding ICP values. Thus, the results show that CO2 reactivity is at least partially determined by CPP and that the impaired CO2 reactivity observed at intracranial hypertension and arterial hypotension may be caused by reduced CPP. PMID- 11426096 TI - Effects of carbon dioxide pneumoperitoneum on cerebral hemodynamics in pigs. AB - Previous studies have shown that laparoscopic interventions are associated with increases in intracranial pressure. However, the consequences on cerebral blood flow (CBF) are unknown. This study investigates the effects of carbon dioxide (CO2) pneumoperitoneum on CBF in pigs. Ten pigs (weight, 20-26 kg) were anesthetized with 1.4% isoflurane and fentanyl (1 microg/kg per minute). Mechanical ventilation (fraction of inspired oxygen = 0.4) was set to maintain normocapnia (end-tidal CO2 tension = 35 mm Hg). Arterial and central venous catheters were placed for measurement of mean arterial blood pressure and central venous pressure. Bilateral internal carotid artery blood flow was measured using two transient time flow probes placed around both carotid arteries (with ligated external carotid arteries). Cortical and subcortical cerebral blood flow was measured using laser Doppler flowmetry. Sagittal sinus pressure was measured via a superior sagittal sinus catheter. After baseline measurements, the peritoneal cavity of the animals was insufflated with CO2 to achieve an intraabdominal pressure of 12-mm Hg. After 10 minutes of stable CO2, pneumoperitoneum measurements were repeated. Increases in central venous pressure (6.3 +/- 2.1 to 11.1 +/- 3.0 mm Hg) and sagittal sinus pressure (8.0 +/- 2.8 to 11.9 +/- 3.0 mm Hg) were noted during CO2 pneumoperitoneum (P < .05). Bilateral internal carotid artery blood flow (46.0 +/- 7.4 vs 47.7 +/- 7.1 mL/100g per minute), cortical CBF (263 +/- 115 vs 259 +/- 158 tissue perfusion units), and subcortical CBF (131 +/- 145 vs 133 +/- 149 tissue perfusion units) did not change during CO2 pneumoperitoneum. The current data show that CO2 pneumoperitoneum increases sagittal sinus pressure without changing CBF. Increases in sagittal sinus pressure are likely related to decreases in cerebral venous drainage caused by increases in intraabdominal pressure. PMID- 11426097 TI - Effects of nimodipine and magnesium sulfate on endogenous antioxidant levels in brain tissue after experimental head trauma. AB - To examine the effects of calcium antagonists nimodipine and magnesium sulfate (MgSO4) on tissue endogenous antioxidant levels, the authors studied superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels in rabbit brain 1 hour after experimental head trauma. Forty New Zealand rabbits were anesthetized and randomly divided into four groups. Group 1 (n = 10) was the sham operated group. Group 2 (n = 10), the control group, received head trauma and no treatment. Group 3 (n = 10) received head trauma and intravenous (IV) 2 microgr/kg nimodipine. Group 4 (n = 10) received head trauma and IV 100 mg/kg MgSO4. Head trauma was delivered by performing a craniectomy over the right hemisphere and dropping a weight of 20 g from a height of 40 cm. In the right (traumatized) hemisphere, SOD and GPx decreased by 57.60% +/- 9.60% and 72.93% +/- 5.51% respectively from sham values. Magnesium sulfate, but not nimodipine, reduced the magnitude of decrease of SOD and GPx to 19.43% +/- 7.15% and 39.01% +/- 7.92% respectively from sham values. In the left (nontraumatized) hemisphere, MgSO4 increased SOD to 42.43% +/ 24.76% above sham values. The authors conclude that MgSO4 treatment inhibited the decrease in SOD and GPx levels in experimental brain injury. PMID- 11426098 TI - Cerebral autoregulation before and after blood transfusion in a child. AB - The authors present the case of an anemic 22-month-old child undergoing lower extremity surgery in whom the lower limit of cerebral autoregulation was shifted to the right. PMID- 11426099 TI - Oropharyngeal swelling and macroglossia after cervical spine surgery in the prone position. AB - An unusual case of massive oropharyngeal swelling and macroglossia occurring after cervical spine surgery performed on a patient in the prone position is described. Possible etiological factors are reviewed, and measures to prevent these complications are suggested. PMID- 11426100 TI - Rapacuronium administration to patients receiving phenytoin or carbamazepine. AB - Patients receiving anticonvulsants such as phenytoin or carbamazepine may be resistant to neuromuscular blocking agents. The authors report the response to rapacuronium bromide (1.5 mg/kg) in two adult patients; one receiving phenytoin and the other receiving carbamazepine. In both patients, there was a delay in achieving maximum blockade; 100% depression of the first twitch was never achieved in the patient receiving phenytoin. Recovery of neuromuscular function was rapid. In the patient receiving phenytoin and carbamazepine respectively, the clinical duration (time to return of T1% to 25% of baseline) was 5 and 9 minutes, the recovery index (T1 25%-75%) was 4 minutes and 3 minutes, and the time to return of T4/T1 to greater than 0.7 was 15 minutes and 18 minutes 40 seconds. As has been reported with other neuromuscular blocking agents of the aminosteroid class, the clinical duration and the recovery index of rapacuronium are shortened in patients receiving either phenytoin or carbamazepine. PMID- 11426101 TI - Dynamic intraoperative kinking of flexometallic tube. AB - A 22-year-old male patient had two episodes of oxygen desaturation with concomitant increase in end-tidal carbon dioxide and airway pressure while undergoing transoral odontoidectomy under fluoroscopy. Dynamic kinking of the flexometallic endotracheal tube from compression by a Dingman retractor was responsible. Fluoroscopic imaging was helpful in confirming the etiology of sudden alteration in the monitoring parameters. PMID- 11426102 TI - Monitored anesthesia care using remifentanil and propofol for awake craniotomy. AB - Adequate analgesia and sedation with adequate respiratory and hemodynamic control are needed during brain surgery in awake patients. In this study, a protocol using clonidine premedication, intraoperative propofol, remifentanil, and labetalol was evaluated prospectively in 25 patients (aged 50 +/- 16). In all but one patient, no significant problems regarding cooperation, brain swelling, or loss of control were noticed, and it was not necessary to prematurely discontinue any of the procedures. One patient, who was uncooperative and hypertensive, became apneic with increasing sedation, and needed a laryngeal mask airway inserted. Patients were hemodynamically stable; elevated systolic blood pressure (>or= 150 mm Hg) was measured infrequently, and there were no events of significant hypotension, tachycardia, or bradycardia. Events of hypoxemia (SAO2 or= 10) to HRV in patients characterized by a worsened neurologic state (GCS < 10). Statistical analysis used the Kruskal-Wallis test, P < .05. To assess whether HRV could predict evolution to brain death, receiver operating characteristic (ROC) curves were generated the day after trauma for Total Power, natural logarithm of high-frequency component of spectral analysis (LnHF), natural logarithm of low frequency component of spectral analysis (LnLF), and root mean square for successive interval differences (rMSSD). Seven patients died between Day 1 and Day 5 after trauma. Six of those had progressed to brain death. In these six patients, at Day 1, Global HRV and parasympathetic tone were significantly higher. Referring to the area under the rMSSD ROC curve, HRV might provide useful information in predicting early evolution of patients with severe head trauma. During the awakening period, global HRV and the parasympathetic tone were significantly lower in the worsened neurologic state group. In conclusion, HRV could be helpful as a predictor of imminent brain death and a useful adjunct for predicting the outcome of patients with severe head injury. PMID- 11426107 TI - Carotid artery disease: should angioplasty be considered an alternative to open carotid surgery? AB - The importance of treating symptomatic high-grade stenosis of the carotid artery is now widely acknowledged. The criteria for nonmedical treatment is continually refined by taking into account the patient's symptoms, age, gender, family history, and treatment-related risk factors. Such refinements have realized importance because the small benefit of invasive treatment can be negated by treatment complications. Improvements in medical treatments such as novel antiplatelet agents, antioxidant, and statin therapy, will ensure that the gain from invasive methods will be further reduced. Surgical (open) carotid endarterectomy (CEA) is the gold standard treatment for high-risk lesions, because it achieves the crucial goal of excising the culprit atheroma, thus providing treatment durability. However, this treatment is highly dependent on surgical standards, and therefore, it may not be beneficial in all centers. In carotid angioplasty (CAP), the atheroma is distended with an endovascular balloon. Because the atheroma is not removed, but the pathologic process is altered, durability cannot be assumed and restenosis remains a significant possibility. In this article, the areas of concern for CEA and CAP will be compared and contrasted. I will address the standards of care used when treating patients with carotid artery disease, and discuss the procedure that should be followed when introducing new treatment options. PMID- 11426108 TI - Angioplasty and stenting is a reasonable treatment option for many patients with atherosclerotic carotid stenosis. AB - A growing number of patients with atherosclerotic stenosis of the carotid artery are being treated endovascularly by percutaneous transluminal balloon angioplasty and vascular stent placement (angioplasty and stenting). However, there is considerable controversy surrounding the use of this procedure. Like many controversial issues in medicine, there are both political and scientific aspects to consider. If all treatment modalities for atherosclerotic carotid stenosis fell within the scope of a single specialty, much of the current hue and cry would be absent. In this short essay, I will outline the scientific issues regarding the treatment of carotid stenosis and provide a rationale for the use of angioplasty and stenting in selected patients. PMID- 11426110 TI - Accurate deployment of vena cava filters: comparison of intravascular ultrasound and contrast venography. AB - BACKGROUND: The increasing use of vena cava filters by trauma surgeons has led to reports of filter placement using intravascular ultrasound (IVUS). Although attractive because of its ease of use and elimination of contrast and radiation, no studies have examined the accuracy of filter placement by IVUS as compared with contrast venography (CV). The purpose of this study was to compare the anatomic information obtained by both techniques during filter placement. METHODS: Twenty-one patients meeting trauma service criteria for filter placement were studied (11 women and 10 men; mean age, 46.8 years). All procedures were performed in the operating room by trauma surgeons. Vascular access was obtained by percutaneous placement of an 8 French sheath in the right femoral vein. CV, IVUS, and bilateral selective renal venography were performed before deployment of a Greenfield filter. Localization and diameter measurements were made in reference to a radiopaque ruler placed on the patient's abdomen. We chose the "best location" for filter deployment as 1 cm below the junction of the lowest renal vein and the vena cava. Measurements by CV and IVUS were compared with the "gold standard" of selective renal venography. RESULTS: As compared with selective renal venography, the difference between best location by CV and IVUS was 16.3 +/- 13.8 mm and 3.7 +/- 5.6 mm, respectively (p = 0.001). In four cases (19%) the CV missed best location by 3 cm or more. CV overestimated the diameter of the vena cava in all cases. Average vena cava diameter was 26.4 +/- 3.3 mm by venography and 20.6 +/- 3.1 mm by IVUS (p < 0.0001). CV incorrectly identified four patients as having vena cava diameters too large (>2.8 cm) for the placement of a Greenfield filter. The two renal vein anomalies (one double left renal vein and one absent left renal vein) were correctly diagnosed by IVUS. CONCLUSION: IVUS is a more accurate method of localizing the renal veins and measuring vena cava diameter for placement of vena cava filters than contrast venography. PMID- 11426111 TI - Spontaneous in contrast to CD95-induced neutrophil apoptosis is independent of caspase activity. AB - BACKGROUND: The influence of caspase inhibitors on spontaneous and on CD95 triggered apoptosis was investigated in neutrophils from healthy volunteers and compared with neutrophils from patients with severe sepsis. METHODS: To further elucidate the mechanisms of neutrophil apoptosis, isolated neutrophils from healthy volunteers (n = 9) were either stimulated with the agonistic anti-CD95 antibody (100 ng/mL) or left unstimulated in the presence or absence of the caspase inhibitors zIETD-fmk (10 micromol/L), zDEVD-fmk (10 micromol/L), or zVAD fmk (20 micromol/L). Apoptosis was determined by measuring DNA fragmentation and Annexin-V binding in FACS, and caspase-3-like activity by DEVD-afc cleavage assay. Results were compared with those from patients with severe sepsis (n = 15). RESULTS: Reduced spontaneous neutrophil apoptosis in patients with sepsis ( 48.7%) was completely restored by incubation with agonistic anti-CD95 antibody (p < 0.05). Inhibition of caspases did not influence spontaneous neutrophil apoptosis in both groups. However, zVAD-fmk reduced anti-CD95 antibody-induced apoptosis in neutrophils from controls by -22.6% (p < 0.05) and in patients with sepsis by -43.1% (p < 0.05). CONCLUSION: These results indicate that spontaneous in contrast to CD95-induced neutrophil apoptosis is independent of caspase activity. PMID- 11426112 TI - Major secondary surgery in blunt trauma patients and perioperative cytokine liberation: determination of the clinical relevance of biochemical markers. AB - BACKGROUND: The aim of this study is to assess the associations between the timing of secondary definitive fracture surgery on inflammatory changes and outcome in the patient with multiple injuries. The study population consists of a series of patients with multiple injuries who were managed using a strategy of primary temporary skeletal stabilization followed by delayed definitive fracture fixation. METHODS: In a prospective cohort study performed at a Level I trauma center, the patients' injuries and operative details as well as immune markers and clinical outcomes were studied. The patients were split into an early secondary surgery group (group ESS, surgery at days 2-4) and a late secondary surgery group (group LSS, surgery at days 5-8). During the posttraumatic course, inflammatory markers (interleukin [IL]-6, tumor necrosis factor-alpha) were determined on a daily basis. Perioperatively, these markers were additionally evaluated at 30 minutes, 7 hours, and 24 hours after initiation of surgery. RESULTS: Secondary surgery on days 2 to 4 was associated with a higher incidence of postoperative organ dysfunction (n = 33 [46.5%]) than secondary surgery on days 5 to 8 (n = 9 [15.7%], p = 0.01). A significant association between the combination of initial IL-6 values > 500 pg/dL plus surgery on days 2 to 4 and the development of multiple organ failure (r = 0.96, p < 0.001) occurred. A correlation between the initial IL-6 values > 500 pg/dL and surgery on days 5 to 8 (r = 0.57, p < 0.07) could not be found. IL-6 also demonstrated a predictive value for the development of multiple organ failure: IL-6 > 500 pg/dL in group ESS, r = 0.96, p < 0.001; IL-6 > 500 pg/dL in group LSS, r = 0.57, p < 0.07. CONCLUSION: According to our data, no distinct clinical advantage in carrying out secondary definitive fracture fixation early could be determined. In contrast, in patients who demonstrated initial IL-6 values above 500 pg/dL, it may be advantageous to delay the interval between primary temporary fracture stabilization and secondary definitive fracture fixation for more than 4 days. In patients with blunt multiple injuries undergoing primary temporary fixation of major fractures, the timing of secondary definitive surgery should be carefully selected, because it may act as a second hit phenomenon and cause a deterioration of the clinical status. PMID- 11426113 TI - Antimicrobial efficacy of external fixator pins coated with a lipid stabilized hydroxyapatite/chlorhexidine complex to prevent pin tract infection in a goat model. AB - BACKGROUND: Pin tract infection is a common complication of external fixation. An antiinfective external fixator pin might help to reduce the incidence of pin tract infection and improve pin fixation. METHODS: Stainless steel and titanium external fixator pins, with and without a lipid stabilized hydroxyapatite/chlorhexidine coating, were evaluated in a goat model. Two pins contaminated with an identifiable Staphylococcus aureus strain were inserted into each tibia of 12 goats. The pin sites were examined daily. On day 14, the animals were killed, and the pin tips cultured. Insertion and extraction torques were measured. RESULTS: Infection developed in 100% of uncoated pins, whereas coated pins demonstrated 4.2% infected, 12.5% colonized, and the remainder, 83.3%, had no growth (p < 0.01). Pin coating decreased the percent loss of fixation torque over uncoated pins (p = 0.04). CONCLUSION: These results demonstrate that the lipid stabilized hydroxyapatite/chlorhexidine coating was successful in decreasing infection and improving fixation of external fixator pins. PMID- 11426114 TI - Impact of recent trends of noninvasive trauma evaluation and nonoperative management in surgical resident education. AB - BACKGROUND: The use of ultrasonography and nonoperative management of solid organ injury has become standard practice in many trauma centers. Little is known about the effects of these changes on resident educational experience. METHODS: We retrospectively reviewed resident evaluation of abdominal trauma and trauma operative experience as reported to the residency review committee between 1994 and 1999. RESULTS: A total of 4,052 patients underwent one or more of three diagnostic modalities. The nontherapeutic laparotomy rate as a result of positive diagnostic peritoneal lavages decreased from 35% to 14%. Although resident operative trauma experience was stable because of increases in operative burns and nonabdominal trauma, the number of abdominal procedures declined. CONCLUSION: Noninvasive diagnostic tests have allowed more rapid trauma evaluation and fewer nontherapeutic laparotomies. As nonoperative experience grows, the opportunity for operative experience decreases. These trends may adversely affect the education of residents and suggest that novel approaches are needed to ensure adequate operative experience in trauma. PMID- 11426115 TI - Factors affecting mortality rates in patients with abdominal vascular injuries. AB - BACKGROUND: Major vessel injury is seen in 5% to 25% of patients admitted to hospitals with abdominal trauma, and this is the most common cause of death in these patients. METHODS: Data on 470 patients with abdominal vascular injuries seen at a Level I trauma center were reviewed retrospectively. RESULTS: The overall mortality rate was 45%. The incidence of various types of trauma were blunt in 51 patients (11%), gunshot wounds in 329 patients (70%), shotgun wounds in 21 patients (4%), and stab wounds in 69 patients (15%). The three vessels with the highest mortality rates were aorta (at and proximal to the renals) (32 of 35 [91%]), hepatic veins and/or retrohepatic vena cava (36 of 41 [88%]), and portal vein (25 of 36 [69%]). The most significant risk factors (p < 0.001) for death were a trauma score of 9 or less, initial operating room (OR) systolic blood pressure (SBP) < 90 mm Hg, final OR core temperature < 34 degrees C, 10 or more blood transfusions in the first 24 hours, and an initial emergency department SBP < 70 mm Hg. Of 120 patients with an initial OR SBP < 70 mm Hg, 103 (86%) died. Of 29 patients with a good response to a prelaparotomy thoracotomy with thoracic aortic cross-clamping (SBP > 90 mm Hg within 5 minutes), 11 (38%) survived. Of the remaining 87 patients, only 6 (7%) survived (p = 0.01). CONCLUSION: Rapid control of bleeding sites (to keep blood transfusions to < 10 units) and urgent correction of hypothermia seem to be the main factors improving survival over which the surgeon has some control. PMID- 11426116 TI - Emergency room thoracotomy for penetrating chest injury: effect of an institutional protocol. AB - BACKGROUND: Emergency room thoracotomy (ERT) can be life saving in patients with penetrating chest injury. A protocol was established at our institution stating that ERT be performed for cases of cardiac tamponade secondary to penetrating chest trauma on patients with vital signs/mentation in the field or on arrival to the emergency room. To validate our protocol, we reevaluated patients undergoing ERT at our institution. METHODS: In our retrospective review, there were 49 patients undergoing ERT over a 6-year period. RESULTS: Survival in patients with vital signs was approximately 50%. Survival in those without was 0%. Compared with the preprotocol data, the number of ERTs declined from 32.2 cases per year to 8.1 cases per year. Overall survival increased from 4% to 20%. Neurologic outcome remained unchanged. CONCLUSION: We believe that the data validate our protocol, and the establishment of a guideline has enabled us to maximize patient survival and minimize exposure risks to our staff. PMID- 11426117 TI - Metabolic and hemodynamic effects of CO2 pneumoperitoneum in a controlled hemorrhage model. AB - BACKGROUND: Intracavity infusion of fibrin sealant-based agents, as a novel modality to control internal bleeding, is associated with an increase of pneumoperitoneum (PP) pressure. The safe limit of such increase has not been well defined in hypovolemic subjects. The purpose of this study was to evaluate the hemodynamic and metabolic effects of increasing PP pressure and to define the limits of carbon dioxide (CO2) insufflation in a controlled hemorrhage rat model. METHODS: Ninety male rats (474 +/- 6 g, 37 degrees +/- 1 degrees C) were anesthetized, and mechanically ventilated. Animals were randomly distributed among 14 groups (n = 6-8) with an increasing amount of blood loss (0, 10, 15, and 17.5 mL/kg) and 15 minutes of CO2 insufflation at 0, 5, 10, and 15 mm Hg starting 15 minutes after hemorrhage, followed by desufflation. Mean arterial pressure (MAP), heart rate, and survival were recorded and arterial and venous blood samples were collected at baseline, at 15 minutes after hemorrhage, after insufflation, and after desufflation procedures to determine arterial blood gases and lactic acid levels. RESULTS: In nonhemorrhaged animals, increasing PP pressure up to 15 mm Hg produced only transient changes in MAP and no increase in lactate level. A moderate hemorrhage (10 mL/kg) limited the safe abdominal pressure to 10 mm Hg with metabolic changes that were restored 15 minutes after desufflation. Higher PP pressure (15 mm Hg) at this hemorrhage level produced a significant decline in MAP (42%, p < 0.001) and progressive metabolic acidosis with a 2.1-fold increase (p < 0.01) in lactate level. The more severe hemorrhage (15 mL/kg) further reduced the limits of PP pressure such that 10 and 15 mm Hg resulted in a progressive decline of blood pressures (52% and 54%, respectively; p < 0.001) and severe metabolic acidosis as manifested by 3.3- and 3.1-fold rises in lactate levels, respectively. In the most severe hemorrhaged animals (17.5 mL/kg), the 50% mortality was primarily determined by the severity of the blood loss and the additional PP at 5 mm Hg had no significant impact. CONCLUSION: The safe limit of PP pressurization with CO2 is dependent on the amount of blood loss. In this mechanically ventilated rat model, increasing the amount of blood loss from 0 to 15 mL/kg reduces the tolerable level of abdominal insufflation pressure from 15 mm Hg to 5 mm Hg. A 5-mm Hg PP pressure appears safe even in the most severely hemorrhaged animals. PMID- 11426118 TI - Routine evaluation of the cervical spine in head-injured patients with dynamic fluoroscopy: a reappraisal. AB - BACKGROUND: The mechanism for clearing the cervical spine in patients with altered mental status remains controversial. Recommendations have ranged from removal of the cervical collar after 24 hours in patients with normal radiographs, to indefinite immobilization in a cervical collar, and recently cervical flexion-extension examinations using dynamic fluoroscopy. The purpose of this study was to evaluate the efficacy and safety of dynamic fluoroscopy flexion extension examinations in identifying ligamentous cervical spine injury and clearing the cervical spine in patients with altered mental status after trauma. METHODS: Patients with a Glasgow Coma Scale score < 13 for greater than 48 hours after admission and normal cervical spine radiographs were candidates for fluoroscopic evaluation. The protocol required visualization of the entire cervical spine, through T1, through full extension and flexion under the direct supervision of a radiologist. Oblique fluoroscopic views were obtained, as necessary, to visualize the cervicothoracic junction. Demographic data, fluoroscopy time, total time per study, true and false positives and negatives, and complications were recorded. RESULTS: From July 1992 through December 1999, fluoroscopic examinations were performed on 301 patients. There were 297 true negative examinations, 2 true-positive examinations (stable injuries), 1 false negative examination, and 1 false-positive examination. The incidence of ligamentous injury identified by fluoroscopy in this study was 2 of 301 (0.7%). Unstable cervical spine ligamentous injuries were identified in only 0.02% of all trauma patients. One patient developed quadriplegia when fluoroscopic evaluation was performed after two protocol violations. CONCLUSION: Unstable cervical spine ligamentous injury without fracture is a rare occurrence. The cervical spine may be cleared after a normal cervical spine series (plain radiograph and computed tomographic scan) as recommended in the 1998 Eastern Association for the Surgery of Trauma guidelines. If dynamic fluoroscopy is to be used, adherence to the protocol, including review of the cervical spine radiographs before fluoroscopy and visualization of the entire cervical spine, C1-T1, is mandatory to ensure patient safety. PMID- 11426119 TI - The management of suspected cervical spine injury in the difficult to evaluate patient. PMID- 11426120 TI - Safety and feasibility of craniectomy with duraplasty as the initial surgical intervention for severe traumatic brain injury. AB - BACKGROUND: Decompressive craniectomy has historically served as a salvage procedure to control intracranial pressure after severe traumatic brain injury. We assessed the safety and feasibility of performing craniectomy as the initial surgical intervention. METHODS: Of 29 consecutive patients undergoing emergent decompression for severe traumatic brain injury with horizontal midline shift greater than explained by a removable hematoma, 17 had traditional craniotomy with or without brain resection and 12 underwent craniectomy. RESULTS: The craniectomy group had lower Glasgow Coma Scale scores at surgery (median, 4 vs. 7; p = 0.04) and more severe radiographic injuries (using specific measures). Mortality, Glasgow Outcome Scale scores, Functional Independence Measures, and length of stay in both the acute care setting and the rehabilitation phase were similar between the surgical groups. CONCLUSION: Despite more severe injury severity, patients undergoing initial craniectomy had outcomes similar to those undergoing traditional surgery. A randomized evaluation of the effect of early craniectomy on outcome is warranted. PMID- 11426121 TI - What is gross hematuria? Correlation of subjective and objective assessment. AB - BACKGROUND: Gross hematuria is a screening test used to help determine the need for radiographic evaluation in patients with blunt trauma. This subjective assessment has not been compared with objective measures, nor has interrater reliability been described. METHODS: We performed a prospective, randomized, controlled study to determine the ability of clinicians to assess gross hematuria. Clinicians were asked to assess samples containing various concentrations of blood diluted in urine for the presence of gross hematuria. RESULTS: Gross hematuria was recognized by more than 95% of clinicians only when samples contained more than 3,500 red blood cells per high-power field. Clinicians' interpretations were independent of profession, specialty, and level of training (p > 0.08). CONCLUSION: Clinicians demonstrate poor sensitivity and interrater reliability identifying gross hematuria by inspection. Clinical studies correlating degrees of hematuria to the need for genitourinary diagnostic evaluation should report both subjective and objective measures to identify a threshold value predictive of renal injury. PMID- 11426122 TI - Hypotensive resuscitation using a polymerized bovine hemoglobin-based oxygen carrying solution (HBOC-201) leads to reversal of anaerobic metabolism. AB - BACKGROUND: Traditional resuscitation regimens have been recently challenged. This study evaluates hypotensive resuscitation with a hemoglobin-based oxygen carrying (HBOC) solution after severe hemorrhage in a porcine model. We hypothesized that HBOC-201 restores tissue perfusion at a lower mean arterial pressure than standard resuscitation fluids. METHODS: Yorkshire swine (55-65 kg, n = 30), were rapidly hemorrhaged to a mean arterial pressure (MAP) of 30 mm Hg, maintained hypotensive for 45 minutes, and randomized into groups. Group I was resuscitated with an HBOC solution to a MAP of 60 mm Hg. Groups II and III were resuscitated to a MAP of 80 mm Hg with lactated Ringer's solution (LR) alone or LR (40 mL/kg) followed by shed blood, respectively. Group IV was resuscitated with shed blood alone to a MAP of 60 mm Hg. Group V received an HBOC solution to a MAP of 50 mm Hg. Hemodynamic variables, Swan-Ganz parameters, blood gas samples, and lactate levels were followed for 5 hours. Data were analyzed by analysis of variance/Duncan multiple range test. RESULTS: There were no significant differences in mortality between any groups. Groups I, IV, and V had lower (p < 0.05) cardiac output, pulmonary artery wedge pressure, and MAP than either group II or group III. Svo2 was significantly lower in the HBOC groups. There were no significant differences in arterial pH or lactate between groups I, III, and IV. Lactate levels, base excess, and arterial pH were significantly worse in the LR-alone and HBOC-50 groups. CONCLUSION: Hypotensive resuscitation with HBOC-201 at a MAP of 60 mm Hg after a controlled hemorrhage in swine provides sufficient tissue perfusion and oxygen delivery to reverse anaerobic metabolism on the basis of global physiologic markers despite continued hypotension, hypovolemia, and low cardiac output. PMID- 11426123 TI - Effect of hydroxyethyl starch on coagulopathy in a swine model of hemorrhagic shock resuscitation. AB - BACKGROUND: Hydroxyethyl starch (HES) has a known dose-dependant effect on coagulopathy. The purpose of this study was to determine the effect of HES on coagulopathy after a period of hemorrhagic shock. METHODS: Anesthetized swine underwent a 15-minute, 40% blood volume hemorrhage (28 mL/kg) and a 1-hour shock period, followed by resuscitation with sham resuscitation (group I); 6% HES, 15 mL/kg (group II); 5% albumin, 15 mL/kg (group III); lactated Ringer's solution, 39 mL/kg, and 6% HES, 15 mL/kg (group IV); and lactated Ringer's solution, 39 mL/kg, and 5% albumin, 15 mL/kg (group V). Coagulation function was measured by bleeding time, prothrombin time, partial thromboplastin time, fibrinogen, platelet count, and thromboelastography. RESULTS: Platelet counts decreased significantly (p < 0.05) in all resuscitation groups except the sham resuscitation group. A significant decrease in platelets, fibrinogen levels, and maximum amplitude on thromboelastography was related to a dilutional effect of the fluid given and not a result of HES at the dose tested. CONCLUSION: The linear dose-related coagulopathic effects of HES when given at moderate doses does not seem to be worsened by prolonged periods of hemorrhagic shock. The coagulopathy seen during resuscitation from hemorrhagic shock seems to be a dilutional effect. PMID- 11426124 TI - Treatment of proximal humeral fractures by intramedullary Kirschner wires. AB - BACKGROUND: This study is a retrospective analysis of a case series of 41 proximal humeral fractures treated by multiple intramedullary Kirschner (K) wires. METHODS: Forty-one proximal humeral fractures in patients aged 17 to 62 years were treated by closed intramedullary K-wires. There were 18 patients with the 2-part fractures, 11 with the 3-part fractures and 12 with the 4-part fractures according to Neer's classification. Patients were placed supine on the table, under general anesthesia. The arm was held vertically up by transolecranon traction, and the C-arm was kept parallel to the ground and opposite to the affected arm. A cortical window was created approximately 1.5 cm proximal to the olecranon fossa, by splitting the triceps. Under fluoroscopic control, multiple K wires were introduced retrograde into the medullary cavity to reach the humeral head across the fracture. The wire tips were kept divergent to provide rotational stability at the fracture site. Postoperatively, the limb was supported in an arm sling. The patients were evaluated for pain, range of motion, strength, anatomic restoration, function of the involved limb, and radiographic union of fracture. RESULTS: An objective evaluation of the results using Neer's criteria showed 92.7% excellent to satisfactory results. Three patients developed painful shoulder-one from the 3-part and two from the 4-part fracture group. One patient from the 4-part group developed avascular necrosis of the humeral head. CONCLUSION: This surgical technique is effective for proximal humeral fractures. It is recommended for routine management of the 2-part and the 3-part fractures. For the 4-part fractures, it can be used in selected, young and active patients. An endoprosthesis is advised for the old, osteoporotic patients. PMID- 11426125 TI - Management of midclavicular fractures: comparison between nonoperative treatment and open intramedullary fixation in 80 patients. AB - BACKGROUND: The aim of this study was to compare the results achieved in two groups of 40 patients, treated for uncomplicated midclavicular fractures. METHODS: Group 1 (mean age, 37.2 years) underwent nonoperative treatment with a figure-of-8 bandage, whereas group 2 (mean age, 30.2 years) underwent open reduction and intramedullary fixation with a 2.5-mm threaded pin. The groups were similar with respect to fracture type. RESULTS: A high rate of complications occurred in group 2, including eight superficial infections, three refractures, two delayed unions with pin breakage, and two nonunions. Return to daily activities was undertaken after an average of 16.7 days from trauma in group 1, and after 40.7 days in group 2 (p = 0.00). Also, return to heavy and sport activities was more rapid for patients treated conservatively: 2.6 months versus 3.2 months (p = 0.014). At follow-up, which averaged 63.7 months, clinical evaluation according to the Constant rating scale did not show significant differences between the two groups. The absolute score averaged 84.8 in group 1 and 82.9 in group 2, whereas the mean relative scores were 94.9% and 95%, respectively. Thirty patients of each group were completely satisfied with the treatment received. The most common cause of dissatisfaction was represented by the unaesthetic appearance of the clavicle, because of subcutaneous bone prominence or dystrophic surgical scars. CONCLUSION: According to our experience, we conclude that nonoperative treatment appears more advantageous than open intramedullary fixation for the management of most midclavicular fractures. PMID- 11426126 TI - Spinal injuries in snowboarders: risk of jumping as an integral part of snowboarding. AB - BACKGROUND: The purpose of this study was to clarify the occurrence rate and characteristics of spinal injuries caused by snowboarding that were sustained at the Okumino skiing area in Gifu Prefecture, Japan, from 1988 to 2000. METHODS: This study was a retrospective review of 13,490 cases of snowboard- or ski related injury treated at Sumi Memorial Hospital over this period. RESULTS: A total of 7,188 patients sustained snowboard-related injuries, and 238 of these had spinal injuries caused by snowboarding (3.3%), whereas 6,302 patients sustained ski-related injuries, and 86 of these had spinal injuries caused by skiing (1.4%). Although there were no significant differences in the difficulty of slope, location of vertebral fracture, or spinal cord injury between snowboarders and skiers, the incidence of transverse process fractures was significantly higher in snowboard-related than in skiing-related injury (p < 0.05). In addition, there was a significantly higher incidence of spinal injury among beginner snowboarders than among beginner skiers (p = 0.04). Furthermore, intermediate or expert snowboarders were more likely to be injured because of jumping than beginners (p < 0.001), whereas about 70% of spinal injuries caused by skiing resulted from a simple fall. CONCLUSION: Spinal injuries sustained while snowboarding are increasing considerably in incidence and are characterized as complex injuries. We must educate young snowboarders of the risk of this sport, to prevent these serious injuries. PMID- 11426127 TI - Survival among injured geriatric patients during construction of a statewide trauma system. AB - BACKGROUND: Patient outcomes are presumed to vary during early implementation of a trauma system because of fluctuations in processes of care. This study estimates risk-adjusted survival for injured geriatric patients during implementation of the Washington State trauma system. METHODS: A presystem (1988 1992) versus early construction phase (1993-1995) retrospective cohort analysis of hospitalized geriatric injured patients in Washington State was conducted. Hospital data were cross-linked to death certificates, providing patient follow up. A Cox proportional hazards model assessed survival to 60 days from hospital admission. RESULTS: A total of 77,136 geriatric patients were assessed. No difference in survival was observed (before vs. after) for all geriatric injured patients. However, among severely injured patients (Injury Severity Score > 15), survival during the implementation phase increased by 5.1% compared with patients admitted during the presystem years (p = 0.03). CONCLUSION: This study demonstrates improved survival for seriously injured geriatric trauma patients during construction of the Washington State trauma system. PMID- 11426128 TI - Motor vehicle crashes in New York State: importance of accounting for emergency department deaths when assessing differences in in-hospital mortality by level of care. AB - BACKGROUND: Motor vehicle crashes (MVCs) are one of the leading causes of death in the nation and in New York State, particularly among younger adult males. It is important to study how to reduce mortality from MVCs. METHODS: Hospitalized victims of motor vehicle crashes in the 1994-1995 New York State Trauma Registry were identified for the study. A statistical model was used to calculate risk adjusted mortality rates for groups of hospitals constituting each level of care (regional trauma center, area trauma center, noncenter). Levels of care were also compared with respect to the location of deaths in the hospital (emergency department, inpatient), and the time between emergency department admission and death for patients dying in the hospital. RESULTS: The risk-adjusted mortality rate for MVCs in patients in regional centers was higher, although not significantly higher (6.91%; 95% confidence interval [CI], 6.18%-7.70%) than for area centers (5.53%; 95% CI, 4.43%-6.82%) or for noncenters (5.83%; 95% CI, 4.70% 7.15%). However, regional centers admitted seriously injured trauma patients from the emergency department much more quickly than other levels of care. Whereas only 18% of all in-hospital deaths occurred in emergency departments of regional centers, the comparable percentages for area centers and noncenters were 39% and 46%, respectively. Also, 43% of all deaths in regional centers occurred within 24 hours of presentation to the emergency department, compared with 15% in area centers and 21% in noncenters. CONCLUSION: Risk-adjusted inpatient mortality rates for victims of MVCs may not yield a fair comparison of performance for different levels of care or for different hospitals because of differences in how quickly emergency department patients are admitted to the hospital. A more equitable way to assess hospital mortality rates may be to include emergency department deaths in addition to inpatient deaths. PMID- 11426129 TI - Extensive traumatic soft tissue loss: reconstruction in severely injured patients using cultured hyaluronan-based three-dimensional dermal and epidermal autografts. AB - BACKGROUND: This report demonstrates the potential of two-stage autologous keratodermal grafting as a starting point for noninvasive reconstruction of extensive traumatic soft tissue defects. METHODS: In three severely injured patients, skin biopsies for cell cultivation were taken. Cultured "neodermis" consisting of cultured autologous fibroblasts grown on biocompatible three dimensional scaffolds made up of benzyl ester of hyaluronan was grafted on conditioned defect areas. After ingrowth of dermal substitutes, transplantation of cultured autologous keratinocytes on hyaluronan-based laser-perforated membranes was performed. Ten days later, a 0.2-mm thin, 1:6 meshed autograft was overlaid. Clinical follow-up, histologic, and immunohistochemical findings were documented. RESULTS: Grafting with cultured autologous fibroblasts revealed a suitable dermal tissue replacement. Epithelialization was evident after transplantation of keratinocytes. Final closure of the defects with "normoelastic" tissue properties was achieved after thin mesh-grafting. CONCLUSION: Preliminary findings with the described method seem to be very promising. As in all fields of tissue engineering, long-term studies and further follow-up are required. PMID- 11426130 TI - Successful use of extracorporeal life support in two cases of posttraumatic adult respiratory distress syndrome. PMID- 11426131 TI - Transdiaphragmatic intercostal hernia: review of the world literature and presentation of a case. PMID- 11426132 TI - Complete upper and lower urinary tract obstruction caused by penetrating pellet injury of the kidney. PMID- 11426133 TI - Bilateral brachial plexus palsy resulting from trauma. PMID- 11426134 TI - Presentation of a missed injury of a metatarsophalangeal joint dislocation in the lesser toes. PMID- 11426135 TI - Benzocaine-induced methemoglobinemia accompanying adult respiratory distress syndrome and sepsis syndrome: case report. PMID- 11426136 TI - Potentially life-threatening complication of pulmonary artery catherization. PMID- 11426137 TI - Re.: Correction of cubitus varus by French or dome osteotomy. PMID- 11426140 TI - Don't duck the evidence. PMID- 11426142 TI - The bane of the aging worker. PMID- 11426144 TI - The use of evoked EMG in detecting misplaced thoracolumbar pedicle screws. AB - STUDY DESIGN: Experimental study performed using an animal model. OBJECTIVES: To determine if EMG responses generated by the electrical stimulation of thoracolumbar pedicle screws could be used to predict the screw position. SUMMARY OF BACKGROUND DATA: Evoked EMG has been used successfully to predict pedicle screw position in the lumbar spine. No data have been published on its effectiveness in the thoracic spine. METHODS: A total of 91 screws were inserted into the pedicles from T8 to L2 in six sheep. Monitoring electrodes were placed into transversus abdominus at three levels, the lower two intercostal spaces, and into psoas. A constant voltage stimulus was applied to a probe inserted into each pedicle, and then to each pedicle screw after it had replaced the probe. The threshold voltage required to evoke EMG activity in the relevant myotome was noted. After monitoring the position of each screw was determined by gross dissection. RESULTS: EMG responses in abdominal and intercostal muscles were successfully evoked by thoracic pedicle screw stimulation. Of the 91 screws, 50 were within the pedicle and required an average voltage of 15.12 V to stimulate an EMG response, compared with the 41 misplaced screws that had an average voltage of 7.63 V (P < 0.0001). Using a threshold of 10 V the technique has a sensitivity of 94% and a specificity of 90%. CONCLUSION: Electrical stimulation of pedicle screws and EMG recording in abdominal and leg muscles in sheep provide a reliable indication of pedicle screw position. This technique can be directly applied to human thoracolumbar surgery, but differences in pedicle size would mean that new threshold voltage criteria would need to be established. PMID- 11426145 TI - In vitro genesis of subaxial cervical unilateral facet dislocations through sequential soft tissue ablation. AB - STUDY DESIGN: In vitro anatomic study investigating the degree of soft tissue disruption required to produce a subaxial cervical unilateral facet dislocation. OBJECTIVES: To develop an understanding of the relative contributions to stability of the subaxial cervical soft tissues and to define an anatomic threshold of injury necessary to produce a unilateral cervical facet dislocation. SUMMARY OF BACKGROUND DATA: The literature at this time is unclear regarding the precise pathomechanics of a cervical unilateral facet dislocation and the required threshold of soft tissue injury necessary for its genesis. Published clinical reports do not make any specific reference to these factors or are unclear in their objectivity. METHODS: Two adjacent vertebra at a time in 10 fresh-frozen subaxial cervical spine specimens (C2-C3 to C6-C7) were transfixed in the coronal plane with 3.5-mm Schanz screws. A steady unilateral vertical distraction force resulting in lateral cervical flexion was applied to these screws as the surrounding cervical soft tissue structures were sequentially ablated. Four experimental models were developed, varying the order of soft tissue disruption. RESULTS: The physiologic coupling of subaxial cervical unilateral distraction and rotation, because of the spatial orientation or inclination of the cervical facet joints, allowed the creation of a unilateral facet dislocation without an additional flexion moment. Disruption of the ipsilateral articular capsule, ligamentum flavum, and more than half of the anulus fibrosus was necessary for the genesis of a unilateral facet dislocation. Disruption of the supraspinous and interspinous ligaments was not necessary but appeared to facilitate or lessen the force required to dislocate a unilateral facet. Disruption of the anterior and posterior longitudinal ligaments and intertransverse ligaments was not necessary to create a unilateral facet dislocation. CONCLUSION: This anatomic study further supports the theory that discontinuity of the anterior and posterior longitudinal ligaments is not necessary for a unilateral facet dislocation to occur. The ipsilateral facet capsule, anulus fibrosus, and ligamentum flavum appear to be the physical soft tissue restraints that need to be disrupted to produce a unilateral facet dislocation. PMID- 11426146 TI - Dynamic cervical plates: biomechanical evaluation of load sharing and stiffness. AB - STUDY DESIGN: An in vitro biomechanical study using a simulated cervical corpectomy model to compare the load-sharing properties and stiffnesses of two static and two dynamic cervical plates. OBJECTIVES: To evaluate the load-sharing properties of the instrumentation with a full-length graft and with 10% graft subsidence and to measure the stiffness of the instrumentation systems about the axes of flexion-extension, lateral bending, and axial torsion under these same conditions. SUMMARY OF BACKGROUND DATA: No published reports comparing conventional and dynamic cervical plates exist. METHODS: Six specimens of each of the four plate types were mounted on ultra-high molecular weight polyethylene simulated vertebral bodies. A custom four-axis spine simulator applied pure flexion-extension, lateral bending, and axial torsion moments under a constant 50 N axial compressive load. Load sharing was calculated through a range of applied axial loads up to 120 N. The stiffness of each construct was calculated in response to +/-2.5 Nm moments about each axis of rotation with a full-length graft, a 10% shortened graft, and no graft. ANOVA and Fisher's post hoc test were used to determine statistical significance (alpha < or = 0.05). RESULTS: The two locked cervical plates (CSLP and Orion) and the ABC dynamic plate were similar in flexion-extension, lateral bending, and torsional stiffness. The DOC dynamic plate was consistently less stiff. The Orion plate load shared significantly less than the other three plates with a full graft. Both the ABC and the DOC plates were able to load share with a shortened graft, whereas the conventional plates were not. CONCLUSIONS: All plates tested effectively load share with a full length graft, whereas the two dynamic cervical plates tested load share more effectively than the locked plates with simulated graft subsidence. The effect of dynamization on stiffness is dependent on plate design. PMID- 11426147 TI - Laminoplasty versus laminectomy and fusion for multilevel cervical myelopathy: an independent matched cohort analysis. AB - STUDY DESIGN: A matched cohort clinical and radiographic retrospective analysis of laminoplasty and laminectomy with fusion for the treatment of multilevel cervical myelopathy. OBJECTIVES: To compare the clinical and radiographic outcomes of two procedures increasingly used to treat multilevel cervical myelopathy. SUMMARY OF BACKGROUND DATA: Traditional methods of treating multilevel cervical myelopathy (laminectomy and corpectomy) are reported to have a notable frequency of complications. Laminoplasty and laminectomy with fusion have been advocated as superior procedures. A comparative study of these two techniques has not been reported. METHODS: Medical records of all patients treated for multilevel cervical myelopathy with either laminoplasty or laminectomy with fusion between 1994 and 1999 at our institution were reviewed. Thirteen patients that underwent laminectomy with fusion were matched with 13 patients that underwent laminoplasty. All patients and radiographs were independently evaluated at latest follow-up by a single physician. RESULTS: Cohorts were well matched based on patient age, duration of symptoms, and severity of myelopathy (Nurick grade) before surgery. Mean independent follow-up was similar (25.5 and 26.2 months). Both objective improvement in patient function (Nurick score) and the number of patients reporting subjective improvement in strength, dexterity, sensation, pain, and gait tended to be greater in the laminoplasty cohort. Whereas no complications occurred in the laminoplasty cohort, there were 14 complications in 9 patients that underwent laminectomy with fusion patients. Complications included progression of myelopathy, nonunion, instrumentation failure, development of a significant kyphotic alignment, persistent bone graft harvest site pain, subjacent degeneration requiring reoperation, and deep infection. CONCLUSIONS: The marked difference in complications and functional improvement between these matched cohorts suggests that laminoplasty may be preferable to laminectomy with fusion as a posterior procedure for multilevel cervical myelopathy. PMID- 11426148 TI - Recurrent laryngeal nerve injury with anterior cervical spine surgery risk with laterality of surgical approach. AB - STUDY DESIGN: A detailed review of anterior cervical fusion procedures from a university-based spine specialty service was completed. Noted were the laterality of approach, number of levels, discectomy or corpectomy, use of instrumentation, and cases of reoperation. OBJECTIVES: The primary purpose of the study is to determine whether there is in fact a greater risk of recurrent laryngeal nerve (RLN) injury with approach on the right or left side. Also evaluated is the risk with corpectomy, reoperative procedures, and instrumentation. BACKGROUND: Anatomic considerations have been used as justification to determine the side of surgical approach. However, few clinical studies have delineated the side of surgical approach in their results. METHODS: A total of 328 anterior cervical spine fusion procedures completed between 1989 and 1999 were reviewed. All speech changes reported were noted throughout follow-up. RESULTS: There were 187 anterior discectomy and 141 corpectomy procedures. There were 21 reoperative anterior fusions. There were 173 procedures completed from the right side and 155 from the left. There were nine patients documented to have dysphonia after surgery. Five had a left-sided approach and four had a right-sided approach. CONCLUSIONS: The incidence of RLN symptoms after surgery was 2.7% (9 of 328). The incidence of RLN symptoms was 2.1% with anterior cervical discectomy, 3.5% with corpectomy (5 of 141), 3% with instrumentation (8 of 237), and 9.5% with reoperative anterior surgery (2 of 21). There was a significant increase in the rate of injury with reoperative anterior fusion. There was no association between the side of approach and the incidence of RLN symptoms. PMID- 11426150 TI - Three-dimensional computed tomography-based, personalized drill guide for posterior cervical stabilization at C1-C2. AB - STUDY DESIGN: Cadaver and preliminary clinical study. OBJECTIVES: To enhance the precision of screw positions for posterior transarticular fixations according to Magerl at C1-C2. SUMMARY OF BACKGROUND DATA: The vertebral arteries are at risk during the Magerl operation and may be damaged in up to 4.1% of cases. Even intraoperative navigation, as often used nowadays, does not provide optimal screw positioning in all patients. METHODS: According to the three-dimensional CT data obtained for every individual cadaver or patient, a template was designed for the posterior course of C2: the template contains a drill guide allowing navigated screw positioning inside the left and right isthmus of C2. For a first series of five cadavers a template with clamps connecting only to the lamina of C2, excluding the spinous process from the interface, was carried out. For a second series of three cadavers the template was connected not only to the lamina but also to the spinous process of C2. Both cadaver series were performed without any fluoroscopic control at surgery. Eventually the technology was applied in two clinical cases. RESULTS: The rotational stability of the template toward the lamina C2 was insufficient in the first series, but for the second series both the entry points and screw trajectories were very satisfactory. CONCLUSIONS: Although the actual experience is limited, the idea of using a template with drill guide might simplify and shorten the surgical act and at the same time enhance the accuracy of C1-C2 transarticular screw positioning. PMID- 11426151 TI - The diagnostic performance of vertebral displacement measurements derived from ultrasonic indentation in an in vivo model of degenerative disc disease. AB - STUDY DESIGN: The diagnostic performance of a newly described variable was assessed in an in vivo model of disc degeneration using a split-pair experimental design. OBJECTIVE: To determine if vertebral displacement measures generated from ultrasonic indentation could distinguish between experimental and control groups of animals. SUMMARY OF BACKGROUND DATA: Few procedures are available that noninvasively assess subcutaneous vertebral mechanics. Information from such a procedure would be of value in determining potential clinical relevance of spinal mechanics with respect to low back pain. METHODS: Eight adolescent pigs underwent endplate perforation surgery to initiate lumbar disc degeneration. After 4 months of recovery, these and eight age-matched controls were assessed by ultrasonic indentation, a noninvasive procedure that quantifies vertebral displacements in the plane of loading-indentation. Each animal then received a facetectomy and was reindented at the same location as confirmed by ultrasonic imaging. Discal materials were removed postmortem for analysis. RESULTS: Degenerative discs exhibited morphologic changes consistent with early degenerative disc disease. Prefacetectomy comparison of vertebral displacement measures between control and experimental animals resulted in sensitivity, specificity, and diagnostic accuracy values of 75.0%, 83.3%, and 77%, respectively. After facetectomy these values increased to 87.5%, 83.3%, and 85%, respectively. These measures of diagnostic performance were comparable or superior to those of existing clinical techniques (invasive or otherwise) used to assess degenerative conditions of the spine. CONCLUSIONS: The results of this study suggest that noninvasive measures of vertebral displacement are clinically significant and possess the additional advantages of being objective and noninvasive. PMID- 11426152 TI - Patients' views on importance and usefulness of plain radiography for low back pain. AB - STUDY DESIGN: Quantitative and qualitative cross-sectional interview study. OBJECTIVES: To investigate how patients who are referred for plain radiography because of low back pain perceive the importance and usefulness of the examination. SUMMARY OF BACKGROUND DATA: Up to 50% of plain radiography examinations for low back pain may be unnecessary based on clinical criteria. However, many patients have great confidence in these examinations. A further exploration of the patients' views may indicate how their needs can be met without unnecessary use of radiography. METHODS: Ninety-nine patients (65 women, 34 men) 14-91 years of age who were referred from Norwegian general practitioners for plain radiography of the lumbosacral spine were asked to rate the examination as slightly/fairly or very important (93 responded). Chi-squared tests were used to evaluate differences in rating according to age, gender, clinical history, and clinical appropriateness of the examination, as determined by comparing information in the referral form with Norwegian (NR) and British (BR) recommendations for use of radiography. Each of the 99 patients also underwent a semistructured interview that was based on questions about importance, usefulness, and reasons for the radiography referral. Answers were categorized and described using a qualitative method (template analysis). RESULTS: Seventy two percent (68 of 93) of patients rated radiography as very important. The proportion was higher for men than women (85% vs. 65%, P = 0.04), higher for those with worsening than those with improving/unchanged symptoms (86% vs. 65%, P = 0.03), and higher for inappropriately than appropriately referred patients (NR: 76% vs. 61%, P = 0.17; BR: 81% vs. 56%, P = 0.01). The qualitative analysis showed that the patients related their views on the importance and usefulness of receiving radiography to seven different issues: symptoms and clinical history, information and advice (especially from health care providers), need for emotional support from the physician, need for certainty and reassurance, need for symptom explanation and diagnosis, reliability of radiography compared with clinical evaluation, and expected practical consequences of the radiologic examination. CONCLUSIONS: The finding that inappropriately referred patients tended to rate their radiography referral as more important than appropriately referred patients indicates that the patient's view may be a substantial barrier to appropriate use of radiography. The study identified seven issues underlying the patients' views on importance and usefulness of receiving radiography. Strategies to prevent unnecessary use of plain radiography for low back pain that address these issues are suggested. PMID- 11426153 TI - Racial differences in the use of lumbar spine radiographs: results from the Veterans Health Study. AB - STUDY DESIGN: We analyzed data from the Veterans Health Study, a longitudinal study of male patients receiving VA ambulatory care. OBJECTIVE: To determine whether clinical differences and/or race account for disparities between white and nonwhite patients in the use of lumbar spine radiographs. SUMMARY AND BACKGROUND DATA: Four hundred one patients with low back pain (LBP) receiving ambulatory care services in four VA outpatient clinics in the greater Boston area were followed for 12 months. METHODS: Participants were mailed the Medical Outcome Study Short Form Health Survey (SF-36) and had scheduled interviews that included the completion of a low back questionnaire, a comorbidity index, and a straight leg raising (SLR) test. Using self-reported racial data, patients were grouped as whites (315 patients) and nonwhites (among whom 22 were black, 4 nonwhite Hispanics, and 1 other race). RESULTS: Nonwhite patients had lumbar spine films more often (13 of 27, 48%) than white patients (87 of 315, 27%)(P = 0.02). Nonwhite patients had higher pain intensity scores than white patients (63 +/- 21 vs. 48 +/- 21, P < 0.01) and were more likely to have radiating leg pain (20 of 27, 76%; compared with 171 of 315, 55%; P = 0.01) than white patients. Nonwhite patients had worse physical functioning (P = 0.01), general health perception (P = 0.05), social functioning (P = 0.02), and role limitations because of emotional problems (P < 0.01). At higher LBP intensity level, nonwhite patients received more lumbar spine films (20 of 27, 74%) than did white patients (155 of 315, 50%)(P < 0.01). Among patients with positive SLR test, nonwhite patients also had lumbar spine films more often (5 of 22, 23%) than white patients (29 of 315, 11%) (P < 0.01). However, after adjusting for multiple clinical characteristics, race was no longer found to be an independent predictor of lumbar spine radiograph use. A positive SLR test remained to be associated with higher radiograph use, whereas better mental health status was associated with lower radiograph use. CONCLUSIONS: There was greater use of lumbar spine radiographs by nonwhite patients compared with white patients. This remained true when patients were subcategorized by severity of LBP or SLR test. However, race had no influence when multiple clinical characteristics of the patients were controlled for simultaneously. This study demonstrates the importance of careful and comprehensive case-mix adjustment when assessing apparent differences in the use of medical services. PMID- 11426154 TI - A longitudinal study of the development of low back pain in an industrial population. AB - STUDY DESIGN: This is a longitudinal study in which industrial workers without chronic low back pain (LBP) were initially assessed with a comprehensive test battery and surveyed every 6 months thereafter for 2 years. OBJECTIVE: To determine factors that may predispose industrial workers who lift over 5000 kg per shift to LBP. SUMMARY OF BACKGROUND DATA: Prospective studies are small in number and often limited in breadth or depth of the test battery, methodologic issues, or investigator expertise. There are no prospective studies that focus on a homogeneous work sample of industrial employees. METHODS: Production workers (n = 149) who volunteered for the 2-year study were assessed using physical measures (e.g., muscular strength, endurance, and flexibility), lifting kinematics (a sagittal plane box lift), and health, lifestyle, and work environment data (paper questionnaires). Follow-up questionnaires were distributed every 6 months for 2 years. RESULTS: Using self-report of LBP as the main outcome measure, eight variables predicted LBP in this sample with a 75% correct prediction rate. Predictor variables included age, thoracic acceleration during the trunk velocity test, median frequency intercept of electromyography of the right L3 erector spinae, quadriceps strength, quadriceps endurance, self-assessment of fitness, having a confidante, and number of medications currently taken. CONCLUSION: Results confirmed the multifactorial nature of low back pain and suggest that personal fitness is an important defense against low back pain, even in manual handling lifting tasks. PMID- 11426155 TI - Effect of age on outcomes of tertiary rehabilitation for chronic disabling spinal disorders. AB - STUDY DESIGN: A prospective cohort study evaluating age as a factor in treatment outcomes for chronic disabling work-related spinal disorders undergoing tertiary rehabilitation. OBJECTIVE: To assess the association between age and objective psychosocioeconomic treatment outcomes for work-related spinal disorders undergoing functional restoration. SUMMARY OF BACKGROUND DATA: As early as the 1950s, a link between age and low back symptoms has been identified in the literature. Several studies have demonstrated that the occurrence of low back pain is positively correlated with age up to about 50-60 years, after which prevalence declines. It has been argued that this pattern is stronger for more severe, chronic back pain cases. Little research attention has been given to age as a factor in treatment outcomes for work-related spinal disorders. METHODS: A cohort of 1052 chronically disabled (none working full-time when starting the rehabilitation program) spinal disorder (CDSD) patients were placed into five groups based on age: Group 1, <25 years (22 +/- 2 years, n = 59); Group 2, 25-34 years (30 +/- 3 years, n = 301); Group 3, 35-44 years (39 +/- 3 years, n = 381); Group 4, 45-54 (49 +/- 3 years, n = 237); and Group 5, > or =55 years (59 +/- 4 years, n = 74). All patients completed a functional restoration program combining quantitatively directed exercise progression with a multimodal disability management approach using psychological and case management techniques. Before the start of the program, and again on completion of the program, all patients received a psychosocial evaluation and were also assessed on a variety of physical motion, strength, aerobic, and functional factors, and a cumulative score was calculated, which aggregates and averages these physical measures. A structured clinical interview examining socioeconomic outcomes (work return, health utilization, recurrent injury, and resolution of financial disputes) was conducted at 1 year after program completion, and at least partial information was obtained from this interview on all consecutive patients in the study. RESULTS: The length of pretreatment disability increased with age (P < 0.001), as well as pretreatment surgery rates (P < 0.002). A Mantel-Haenzel chi2 test for linear trend across age groups revealed that the percentage of patients who returned to work declined progressively from 100% in Group 1 to 69% in Group 5 (P < 0.001). The same linear trend was revealed for the percentage of patients retaining work at the end of 1 year (98-62%, P < 0.001). Older workers returned to the same job and/or the same employer (P < 0.005). Younger workers found different jobs with new employers (48-11%, P < 0.0001). Analysis of variance for the disability questionnaire revealed that after treatment subjective disability progressively increased (while improvements decreased) from Group 1 to Group 5 (P < 0.01). The cumulative physical score variable, even normalized for age, was progressively lower from Group 1 to Group 5, both at pretreatment (P < 0.03) and post-treatment (P < 0.02). CONCLUSIONS: The present study represents the first large-scale examination of the association between age and treatment outcomes for a work-related CDSD population. Age is significantly and linearly related to pretreatment duration of disability and frequency of pretreatment surgeries. After rehabilitation treatment there is a linear decrement in both work return and work retention, so that younger patients are far more likely to return and hold work after functional restoration. Older workers who go back to work are much more likely to return to the same employer and do the same job, or to become self-employed. Calculation of odds ratios revealed that patients >55 years are 5.68 times more likely to return to the same job and employer, relative to those <25 years; those >55 years were also approximately three times more likely than those <25 years to return to the same employer, compared with those <25 years. Younger workers are far more likely to take a different job and/or seek a new employer. Older patients are also somewhat more likely to seek a new health provider to deal with perceived residual pain/disability. This trend coincides with their higher post-treatment subjective disability levels, and their lower cumulative physical scores both prerehabilitation and postrehabilitation. However, age does not affect additional surgery rates, subsequent injuries, or delays in settling financial disputes. PMID- 11426156 TI - Early experiences with image-guided transoral surgery for the pathologies of the upper cervical spine. AB - STUDY DESIGN: Technical note. OBJECTIVES: Three years of convincing experience with cranial neuronavigation suggested the application of the cranial software and registration method for the transoral access to the C1-C2 vertebrae. BACKGROUND DATA: The C1-C2 vertebrae are located in close vicinity to the cranial base. If the intersegmental movements of the C0-C1/C1-C2 segments are prevented with HALO fixation, the upper cervical spine can be considered as a caudal part of the skull base and included in the extended navigation space of the skull. METHODS: Three patients were selected for navigation-assisted transoral odontoidectomy. Before surgery the patients were fixed and scanned in a HALO device. The fiducials were attached supraorbitally and to both mastoids, determining a wide registration area and allowing the caudal extension of the navigation space. The BrainLAB VectorVision navigation system was used in cranial mode during the operations. RESULTS: Neuronavigation and fluoroscopy-controlled transoral surgery were performed with success in all three cases. The registration accuracy was 1.5, 2.7, and 3.1 mm. CONCLUSION: Image guidance during transoral exposure of the upper cervical spine offered excellent three dimensional guidance on the ventral surface of the craniocervical junction, allowing a safer, more controlled surgery. As the targets of the transoral spinal surgery are fixed bony and ligamentous structures, no shifting occurs and continuous high navigation accuracy can be achieved. The use of the navigation can reduce the significance of the intraoperative fluoroscopy, diminishing the radiograph load of the patient and the operating room team. PMID- 11426157 TI - Hemivertebra in monozygotic twins. AB - STUDY DESIGN: A case report. OBJECTIVES: To report and discuss a case of thoracic hemivertebra in monozygotic twins. SUMMARY OF BACKGROUND DATA: The etiology of congenital scoliosis remains unclear. Both a genetic basis and environmental influences have been postulated. A few isolated reports of discordant spinal anomalies in twins exist. METHODS: Clinical examination and plain radiograph evaluation were performed on female monozygotic twins with congenital scoliosis secondary to thoracic hemivertebra. RESULTS: Congenital thoracic scoliosis was documented in both of a set of monozygotic twins. CONCLUSIONS: To the authors' knowledge, this is the first report of similar congenital vertebral abnormalities in identical twins in the English literature. PMID- 11426158 TI - Isolated cervical juvenile xanthogranuloma in childhood. AB - STUDY DESIGN: This is a report of an exceptional case of isolated cervical juvenile xanthogranuloma in a child. OBJECTIVES: This case report draws attention to the fact that isolated xanthogranuloma of the central nervous system should be considered among possible diagnosis of subdural extramedullary spinal masses in children and young adults. SUMMARY AND BACKGROUND DATA: Isolated juvenile xanthogranuloma of the central nervous system is extremely rare. When located in the spinal canal it behaves like any extramedullary mass-occupying lesion. MRI depicts the tumor's association with adjacent structures. In cases in which a subtotal surgical removal was possible, radiotherapy has been indicated. METHODS: A three-year-old girl presented severe pain in the right shoulder and spastic tetraparesis. The MRI showed an intradural extramedullary mass homogeneously enhancing after DTPA-gadolinium infusion. Complete surgical removal of the tumor was performed through open-door laminoplasty. RESULTS: The child was pain free immediately after the surgical removal of the tumor. A gradual complete recovery of the neurologic deficits followed. Open-door laminoplasty provided sufficient operative space, and it minimized the impact on the growing spinal column. CONCLUSIONS: Isolated juvenile xanthogranuloma does not show any predilections of localization inside the central nervous system. Both intracranial and spinal juvenile xanthogranulomas appear isointense in MRI and enhance homogeneously with gadolinium. Whenever possible, total surgical removal alone seems to be curative. Otherwise, a subtotal removal of the tumor might be followed by radiotherapy. Immunohistochemical tests ensure the diagnosis. PMID- 11426159 TI - Foramen magnum syndrome secondary to calcium pyrophosphate crystal deposition in the transverse ligament of the atlas. AB - STUDY DESIGN: This report illustrates two rare cases of foramen magnum syndrome caused by a retro-odontoid mass in which calcium pyrophosphate dihydrate crystals were found. OBJECTIVES: To analyze the preoperative studies and the diagnostic criteria and to discuss the surgical treatment. The present cases will be compared with previous ones described in the literature. SUMMARY OF BACKGROUND DATA: Deposition of calcium pyrophosphate dihydrate crystals occurs into the fibrous and hyaline cartilage of the joints and intervertebral discs of the spine. Half of the patients known to have a chondrocalcinosis had asymptomatic calcification in the odontoid region. Ten patients were published in the literature as having a spinal cord syndrome secondary to calcium pyrophosphate dihydrate deposition in the odontoid region. METHODS: In both cases the preoperative studies were analyzed, and the retro-odontoid mass was resected and histologically examined. Both had an anterior transoral approach and have been followed for 1 year. RESULTS: According to the preoperative radiographs the diagnosis was suspected and confirmed histologically. Transoral approach was done with no need in either case for a posterior stabilization. CONCLUSIONS: Compression of the spinal cord by calcium pyrophosphate dihydrate deposition may occur. The preoperative diagnosis may be highly suspected after radiographic study and histologically confirmed. Transoral resection is the treatment of choice. Posterior stabilization should be considered only in cases of craniovertebral instability. PMID- 11426161 TI - Imaging corner: unknown case. Diagnosis and discussion: atlanto-occipital dislocation. PMID- 11426162 TI - Re: training primary care physicians to give limited manual therapy for low back pain: patient outcomes. PMID- 11426163 TI - Re: progressive rotational dislocation in kyphoscoliotic deformities. PMID- 11426164 TI - Re: a radiostereometric analysis of the movements of the sacroiliac joints in the reciprocal straddle position. PMID- 11426165 TI - Load sharing within a human lumbar vertebral body using the finite element method. AB - STUDY DESIGN: A finite element parametric study was performed to investigate the structural roles of the vertebral cortical shell and the trabecular centrum. OBJECTIVES: To address the debated issue of the relative load-carrying role of the vertebral cortical shell. SUMMARY OF BACKGROUND DATA: Several experimental and computational studies have been aimed at quantifying the load-carrying roles of the human vertebral cortical shell and trabecular centrum. These studies, however, have supported no consensus. METHODS: A finite element model of three lumbar vertebral bodies was developed to predict the fraction of the total compressive load acting on the lumbar vertebral body, under two different loading conditions, that was supported by the cortical shell. Parametric variations in vertebral material and geometric properties were examined to determine how this fraction was influenced by such changes. RESULTS: The fraction of the compressive load supported by the cortical shell was found to be strongly dependent on the distance from the endplate, increasing from about 34% at either endplate to approximately 63% at the midtransverse plane. This fraction was independent of the loading characteristics, proportional to the properties of the cortex, and inversely proportional to the modulus of the centrum. Additionally, the cortical shell force fraction was affected significantly by changes in the overall vertebral geometry. CONCLUSIONS: Our findings indicate that the structural dominance of the cortical shell and centrum alternate depending on the location within the vertebral body. However, as age-related bone loss progresses, the load carrying role of the cortical shell could increase significantly. PMID- 11426166 TI - A biomechanical comparison between anterior and transverse interbody fusion cages. AB - STUDY DESIGN: Human cadaveric lumbar spines underwent placement of threaded fusion cages (TFCs) in either an anterior or transverse orientation. Spines underwent load testing and angular rotation measurement in the intact state, after diskectomy, after cage placement, and after fatiguing. Angular rotations were compared between cage orientations and interventions. OBJECTIVE: To determine which cage orientation resulted in greater immediate stability. SUMMARY OF BACKGROUND DATA: There has been extensive biomechanical study of interbody fusion cages. The lateral orientation has been increasingly used for intervertebral fusion, but a direct biomechanical comparison between cages implanted either anteriorly or transversely in human cadaveric spines has not been performed. METHODS: Fourteen spines were randomized into the anterior group (anterior diskectomy and dual anterior cage placement) and the lateral group (lateral diskectomy and single transverse cage placement). Pure bending moments of 1.5, 3.0, 4.5, and 6.0 Nm were applied in flexion, extension, lateral bending, and axial rotation. Load testing was performed while intact, after diskectomy, after cage placement, and after fatiguing. Angular rotation was compared between anterior and lateral groups and, within each group, among the different interventions. RESULTS: Segmental ranges of motion were similar between spines undergoing either anterior or lateral cage implantation. CONCLUSIONS: These results demonstrate few differences between angular rotation after either anterior or lateral TFC implantation. These findings add to data that find few differences between orientation of implanted TFCs. Combined with a decreased risk of adjacent structure injury through a lateral approach, these data support a lateral approach for lumbar interbody fusion. PMID- 11426167 TI - Radiofrequency medial branch neurotomy in litigant and nonlitigant patients with cervical whiplash: a prospective study. AB - STUDY DESIGN: The efficacy of radiofrequency medial branch neurotomy to treat cervical zygapophysial joint pain from whiplash was compared prospectively in litigants and nonlitigants. OBJECTIVES: 1) To assess the effect of monetary gain on treatment of zygapophysial joint pain in cervical whiplash. 2) To determine whether radiofrequency medial branch neurotomy is effective treatment for whiplash. SUMMARY OF BACKGROUND DATA: The influence of litigation on treatment outcome is a subject of controversy in both the medical and legal professions. This is the first study to examine this issue in a prospective manner using a previously proven diagnostic and therapeutic method. METHODS: Sixty patients with cervical whiplash who remained symptomatic after 20 weeks of conservative management were referred for radiofrequency cervical medial neurotomy. The patients were classified as litigant or nonlitigant based on whether the potential for monetary gain via litigation existed. Each group underwent identical evaluation and treatment. Patients were observed for 1 year. Visual analogue scores and self-reported improvement were obtained before, immediately after, and 1 year after radiofrequency cervical medial neurotomy. RESULTS: Forty six patients completed the study. The overall reduction in cervical whiplash symptoms and visual analogue pain scores were significant immediately after treatment (nonlitigants vs. litigants: 2.0 vs. 2.5, P = 0.36) and at 1 year (nonlitigants vs. litigants: 2.9 vs. 4.0, P = 0.05). One-year follow-up scores were higher than immediate post-treatment scores (nonlitigants vs. litigants: 2.5 vs. 3.6). The difference between litigants and nonlitigants in the degree of symptomatology or response to treatment did not reach significance. CONCLUSIONS: These results demonstrate that the potential for secondary gain in patients who have cervical facet arthropathy as a result of a whiplash injury does not influence response to treatment. These data contradict the common notion that litigation promotes malingering. This study also confirms the efficacy of radiofrequency medial branch neurotomy in the treatment of traumatic cervical facet arthropathy. PMID- 11426168 TI - Radiographic spinal profile changes induced by cage design after posterior lumbar interbody fusion preliminary report of a study with wedged implants. AB - STUDY DESIGN: A prospective comparative radiographic study between two geometrically varying implants utilized in single-level posterior lumbar interbody fusion (PLIF) was performed. OBJECTIVE: The aim of this study was to determine the influence of 4 degrees wedged cages on postoperative lateral lumbar spinal profile. SUMMARY OF BACKGROUND DATA: The biomechanical and clinical importance of realigning the sagittal lumbar profile in surgical management of spinal instabilities is known. Wedged cages are therefore increasingly attaining popularity in PLIF. As yet the significance of wedged implants on postoperative sagittal spinal profile has not been assessed. METHODS: Forty patients were randomly assigned to two equal-sized groups. In one group rectangular cages and in the second group cages with a wedged design and an inclination of 4 degrees were implanted. Quantitative assessment of the lumbar spinal profile on standing neutral lateral radiographs was performed before surgery as well as 6 weeks and 12 months after surgery. Results were statistically compared. RESULTS: A significant improvement of lumbar sagittal profile after use of 4 degrees wedged compared with nonwedged cages was not found. The greatest effect on lumbar profile and segmental lordosis was observed in fusion of segment L4-L5 with 4 degrees wedged cages. CONCLUSIONS: These results show that normal sagittal alignment after single-level lumbar fusion can be achieved with rectangular and 4 degrees wedged cages. Although results after utilization of 4 degrees wedged cages do not significantly differ, these implants offer the surgeon one more sizing variation with which physiologic lumbar lordosis may be attained. The biomechanical implications of wedged implants on the rigidity of a fused segment remain to be analyzed. PMID- 11426169 TI - Bilateral phrenic nerve palsy as a complication of anterior decompression and fusion for cervical ossification of the posterior longitudinal ligament. AB - STUDY DESIGN: A case report of bilateral phrenic nerve palsy as a complication of anterior decompression and fusion for cervical ossification of the posterior longitudinal ligament (OPLL). OBJECTIVES: To present a case of a rare and serious complication of cervical spinal surgery and to investigate its cause. SUMMARY OF BACKGROUND DATA: There have been a number of reports of phrenic nerve palsy after cardiac surgery, but the authors have found no previous description of this complication related to spinal surgery. METHODS: The authors describe the clinical presentation and management of a case of bilateral phrenic nerve palsy subsequent to the surgery for cervical OPLL. Also, the literature is reviewed concerning surgical approaches for the treatment of OPLL and the occurrence of phrenic nerve palsy subsequent to any form of therapy. RESULTS: Bilateral phrenic nerve palsy occurred after anterior decompression and fusion for cervical OPLL. Bilateral phrenic nerve palsy was diagnosed radiographically: postoperative chest radiograph showed bilateral laxity of the diaphragm. Movement of the bilateral diaphragm appeared 3 weeks after surgery. The patient successfully returned to normal daily life after ventilatory support for 3 months, although nocturnal oxygen support was still necessary at the latest follow-up, 3 years after surgery. The possible causes of this complication include bilateral C4 nerve root stretching, iatrogenic injury of the gray matter in the ventral horn, alteration of blood circulation related to spinal edema, or re-impingement on the spinal cord at the cranial part of the decompression site. CONCLUSIONS: Bilateral phrenic nerve palsy occurred after anterior decompression and fusion for cervical OPLL. Bilateral phrenic nerve palsy should be kept in mind as a serious complication of spinal surgery. It should be considered when patients unexpectedly fail to wean from the ventilator after surgery. PMID- 11426189 TI - Galantamine (reminyl) for Alzheimer's disease. PMID- 11426190 TI - Pegylated interferon (PEG-Intron) for chronic hepatitis C. PMID- 11426191 TI - A new snake antivenom. PMID- 11426192 TI - [Prevalence of atherosclerosis in renal transplant recipients]. AB - BACKGROUND: Cardiovascular diseases are a major cause of morbidity and mortality after kidney transplantation. Renal transplant recipients have in fact accelerated atherosclerosis because of prolonged hemodialysis, arterial hypertension, dyslipidemia and other vascular risk factors. Studies concerning carotid and lower limb atherosclerotic changes in renal transplanted patients are at present scant. METHODS: In the present study the prevalence of carotid and iliac-femoro-popliteal atherosclerosis was evaluated by color flow Doppler in 99 patients 64 +/- 41 DS months after renal transplantation. Statistical analysis was used to correlate the presence and grade of atherosclerotic lesions with vascular risk factors and other clinical-laboratoristic parameters of the patients. RESULTS: The overall incidence of atherosclerotic lesions in the population studied was 54.5%; 46.5% of patients presented atherosclerotic plaques at carotid district, 43.4% at lower limb arteries and 35.4% at both districts. Two of these patients were symptomatic; one patient affected by claudicatio intermittens was submitted to conservative therapy, whereas another patient with abdominal aortic aneurysm underwent to surgical treatment. Univariate analysis showed a significant difference between males and females for lower limb atherosclerotic lesions (p=0.0013), whereas no correlation was found between carotid lesions and sex, or between smoke and atherosclerosis. Significant correlations were found by multiple correlations analisys between: the grade of atherosclerotic lesions in both districts and the patient age (p<0.0001); the grade of carotid atherosclerotic plaques and duration of pretransplantation dialysis (p<0.01); the grade of lower limb atherosclerosis and respectively glicemic (p<0.01), hematocrit (p<0.01), potassium (p<0.002) values, systolic blood pressure (p<0.02). CONCLUSIONS: Relevant rates of renal transplant recipients have carotid and peripheral atherosclerotic lesions. The study of aorto-iliac and lower extremity vascular atherosclerosis have important implications for the transplanted kidney functionality. Color flow Doppler represents a sensitive method for the follow-up examination of these patients. PMID- 11426193 TI - [Silent ischemia and ventricular arrhythmias in essential hypertension]. AB - BACKGROUND: Silent ischemia episodes rate is 4-5% among over fifty aged people. Patients affected by hypertension have higher coronary artery disease rate than people with normal blood pressure. An increased mortality is present among patients affected by essential hypertension, especially if affected by silent ischemia and /or ventricular arrhythmias. In all previous study about hypertension, the inclusion criterion was a diastolic blood pressure >95 mmHg, that is a low selective one. The aim of study is to evaluate ventricular arrhythmias rate, in hypertensive patients, without pharmacological therapy, and diagnosed by 24 hours ambulatory blood pressure monitoring (ABPM), so using a more selective criterion than WHO rules. METHODS: 128 consecutive patients with hypertension diagnosis by WHO rules, were screened for 24 hours ambulatory blood pressure measurement (ABPM); 85 of them (66.4%) had 24 hours mean blood pressure >135/85 mmHg. These 85 patients were screened for M-mode, B-mode echocardiography and 24 hours electrocardiogram monitoring by Holter. RESULTS: Sixty patients (70.6%) were affected by left ventricular hypertrophy, 25 were free (29.4%) According to the Lown and Wolf classification for ventricular arrhythmias 20 patients (23.5%) had a Grade I arrhythmia, 5 (5.9%) had a Grade II, 4 (4.7%) had a Grade III, 9 (10.6%) had a Grade IV A, 20 (23.5%) had a Grade IV B, 12 (14.2%) had a Grade V and 15 patients (17.6%) were free from premature ventricular complexes. 40 patients (47%) had one or more ST depression episodes longer than 60 . The range of episodes number is 1-22, mean 6.8; their duration range is 1-16 minutes, mean 7.6 minutes. In our study, left ventricular hypertrophy correlate significantly with arrhythmia Lown score, r=0.552 for p<0.0001 and also with silent ischemia as ST depression r=0.51, p<0.004. The correlation, between arrhythmia score and ST depression, r=0.042, p<0.021 is not highly significant. CONCLUSIONS: The conclusion is drawn that using a more selective criterion for the diagnosis of hypertension, it is possible to identify patients affected by a more severe stage of disease, and detect them for primary prevention of coronary events. PMID- 11426194 TI - [Elective and emergency surgical treatment of popliteal aneurysms. Indications and results]. AB - BACKGROUND: The aim of this paper is to evaluate the long term results related to surgical technique and to prosthetic material in planned and emergency conditions. METHODS: From January 1990 to December 1999, fourty-five patients with popliteal aneurysms were observed. Eighteen patients (40%) were asymptomatic; eleven (24.4%) suffered from claudicatio; six had an acute ischemia (13.3%) and four presented (8.9%) clinical signs of rest pain; in four cases (8.9%) symptoms were related to venous compression and to rupture in two (4.4%). Diagnosis was obtained by ultrasounds, angiography and CT-scan. All patients were operated on and in four cases (8.9%) the procedure involved both legs. The prosthetic material was reversed autologous saphenous vein (ASV) in 30 patients (61.2%), PTFE-EXS-TW in 16 (32.6%), homologous vein in one (2%), composite graft in one (2%), and a Dacron in the last one (2%). RESULTS: Six patients died for causes not related to the operation and eight bypasses (18.2%) became occluded. Amputation was needed in two patient, in relation to late bypass occlusion. In one cases an amputation was carried out for acute thrombosis of the contralateral aneurysm which was not treated for patients refusal. The long term patency rate was 81.8%. CONCLUSIONS: Popliteal aneurysms must be considered for reconstructive surgery, also when asymptomatic and with a diameter over 2 cm. The routinely use of the ASV improves the long-term patency rate. Best long term results are obtained in elective surgery. PMID- 11426195 TI - The importance of anatomical variants of the sapheno-femoral junction in lower limb varicose vein surgery. AB - BACKGROUND: In the surgical strategy of lower limb varicose veins, the fundamental objective is to prevent relapses. For this purpose, after having examined the international literature on the most frequent relapse causes, the authors emphasize that erroneous diagnosis, varicose disease natural development and incorrect surgical treatment are very important. METHODS: They report their experience about the sapheno-femoral junction anatomic variants that have been verified at the surgical table on 532 consecutive patients operated on for lower limb varicose veins. Of 532 patients 449 (84.4%) patients were women and 83 (15.6%) were men; all of them were, on average 49+/-26 years old. They were all with derivative from sapheno-femoral junction incontinence varicose veins that can be classified according to CEAP (C2-3-4/Ep/As 2-3/Pr). On these patients, 430 (80.8%) patients have had an operation of internal saphena long stripping; 65 (12.3%) patients have had an operation of saphena short stripping and 37 (6.9%) patients have had an operation of internal simple rossectomy. RESULTS: The authors have inspected all collateral veins of the sapheno-femoral junction and they have noticed 135 variants (25.2%) that are very important for the surgeon and, among these variants, they have discovered some collaterals, that flow under the sapheno-femoral junction (about 4%), are particularly insidious and they are the cause of sure relapse if they are ignored. CONCLUSIONS: The purpose of the work is to emphasize the importance of knowing sapheno-femoral junction changeability that is often the cause of relapse after surgical treatment. PMID- 11426196 TI - Preparation of the internal mammary artery with the IMAG Fogarty balloon catheter. AB - BACKGROUND: Demonstrate the augmentation of flow of the internal mammary artery (IMA) after inflation of the graft with an atraumatic balloon catheter (Fogarty IMAG balloon catheter) before performing the anastomosis IMA-coronary artery, in coronary artery bypass operated patients. METHODS: In 1992 and 1993 a group of 87 coronary artery bypass patients underwent dilatation of the IMA with the Fogarty balloon catheter. The flow of the IMA was measured during 1 minute before and after balloon dilatation, prior to go on cardiopulmonary bypass. RESULTS: In all patients but one the flow increased after balloon dilatation. The mean free flow before dilatation was 52.8 ml/min (range 16-120 ml/min) and after balloon dilatation 122.6 ml/min (range 36-380 ml/min). The flow increased more than two folds which was of statistical significance (p<0.001). The hospital mortality was 6.8% (6 patients) not related with the mechanical manipulation of the arterial graft. CONCLUSIONS: Balloon dilatation of IMA is an alternative in cases in which the IMA graft presented with: 1) low free flow, 2) small size and 3) arterial spasm. The IMAG balloon catheter has to be properly adjusted and the shear force regulated at 30 grams. PMID- 11426197 TI - Caval thrombosis in a young athlete. AB - We here describe a clinical case of caval thrombosis of sudden and unexpected onset, in an athlete without signs of venous insufficiency. Pre-existing caval hypoplasia was observed through imaging examinations. Following a review of the literature, the authors interpret the pathophysiological mechanism of thrombosis as such: a combination of intense physical activity and the malformation led to an overwhelming venous flow and a consequent congestion. Caval thrombosis distal to the malformation is an absolute indication to lifelong oral anticoagulant therapy. PMID- 11426198 TI - [Simultaneous urokinase perfusion in renal artery and vein in a case of renal vein thrombosis]. AB - We present he case of a young man with nephrotic syndrome, caused by membranous glomerulonephritis, who developed renal vein thrombosis with extension to the inferior vena cava is presented. Renal vein thrombosis was diagnosed by echo Doppler and confirmed by angio-CT scan. At the hospitalization the patient presented a severe left flank pain, edema of the lower limbs and painful left testicular tumefaction. The treatment consisted of: 1) systemic anticoagulation with sodic heparin, 2) placement of temporary vena cava filter through the right jugular vein, 3) direct thrombolysis into endocaval thrombus with early lysis of thrombus, and 4) renal thrombolysis with selective simultaneous renal artery and renal vein infusion of urokinase. Angiography performed after 24 hours of loco regional thrombolysis showed complete lysis of renal thrombus; clinically there was a regression of left flank pain. We conclude that, face to renal vein thrombosis, thrombolytic treatment with simultaneous renal artery and renal vein perfusion is mandatory. Furthermore it is very important, in presence of caval extension of renal thrombus, to place a temporary vena cava filter before starting thrombolysis, considering the high risk of pulmonary embolism related to this pathology. PMID- 11426199 TI - [Surgical treatment of atrial thrombosis. A transesophageal echocardiogram study]. AB - Transesophageal echocardiogram is the procedure of choice in the evaluation of masses located in the cardiac chambers. In this paper three cases of atrial thrombosis are presented: a) a free floating ball thrombus in the left atrium in a patient with moderate mitral stenosis; b) three thrombotic masses adherent to a central venous catheter; c) three thrombotic masses attached to Chiari s network in a patient operated for an ostium secundum atrial septal defect one year before. In all cases transesophageal echocardiogram has played a key role in the diagnosis. Medical therapy has been inadequate and so surgical removal has been performed. Unsuccessful medical therapy, a new episode of thrombosis, intracardiac permanent catheter infections and high risk of systemic and pulmonary embolism are indications for thrombectomy. PMID- 11426200 TI - [Formation and development of angiology units]. PMID- 11426201 TI - ASORN's history: a truly amazing group of people accomplishing amazing things. American Society of Ophthalmic Registered Nurses. PMID- 11426203 TI - Psychosocial implications of clinical trials on patients with age-related macular degeneration and pathologic myopia as seen in the photodynamic therapy trials. AB - Photodynamic therapy is a combination of the systemically injected photosensitizing drug, verteporfin, and the subsequent exposure of the affected retina to a low-beam diode laser. Eligible participants in the photodynamic therapy clinical trial were those with "wet" cases of age-related macular degeneration with subfoveal, predominantly classic lesions. The expected outcome of the treatment is to preserve vision, not to restore lost vision. Study participants are frustrated in their performance of daily activities and often have unrealistic expectations of photodynamic therapy treatment outcomes (eg, improved vision). Frustration and fear are evident at visits during which their actual vision loss is measured. One of our goals is to help the patients have realistic expectations of treatment outcomes. PMID- 11426204 TI - Photodynamic therapy: the nurse's role. AB - Patients with neovascular (wet) age-related macular degeneration, the leading cause of blindness in persons older than 50 years in developed countries, experience severe visual loss due to choroidal neovascularization, the growth of abnormal vessels under the retina. Photodynamic therapy is a new treatment modality that combines an intravenous injection of a photosensitizing drug, such as verteporfin, and nonthermal laser light application, which destroys choroidal neovascularization without damaging normal surrounding tissue. It has led to new challenges for nurses working in the ambulatory laser setting, including medication preparation, drug administration, and education regarding precautions and potential systemic side effects of photodynamic therapy. PMID- 11426205 TI - Sexuality. PMID- 11426206 TI - Edna Ashy. PMID- 11426208 TI - Corneal iron deposits after refractive corneal procedures. PMID- 11426209 TI - A-scan (amplitude ultrasonography) measurement. PMID- 11426210 TI - Communication: an important assessment and teaching tool. AB - Communication, as a process, is essential to adequate nursing care; language is a key factor. This article reviews the aspects of language that impair, and those that improve, communication in the health care setting. Standards are cited, along with recommendations for developing optimal communication skills within nursing's professional practice area. This includes the importance of using interpreters when language is a barrier. PMID- 11426211 TI - From CaM kinase to cognition. PMID- 11426212 TI - To burst, or rather, not to burst. PMID- 11426213 TI - Seeing the forest but not the trees. PMID- 11426214 TI - A new form of feedback at the GABA(A) receptor. PMID- 11426215 TI - A chemokine-glutamate connection. PMID- 11426216 TI - Seeing is not perceiving. PMID- 11426217 TI - Boundaries on motion integration. PMID- 11426219 TI - A new developmental compartment in the forebrain regulated by Lunatic fringe. PMID- 11426220 TI - Spine motility with synaptic contact. PMID- 11426221 TI - Fear conditioning and LTP in the lateral amygdala are sensitive to the same stimulus contingencies. PMID- 11426222 TI - Recovery of binocular responses by cortical neurons after early monocular lesions. PMID- 11426223 TI - Cortical reorganization in motor cortex after graft of both hands. PMID- 11426224 TI - Does the brain model Newton's laws? PMID- 11426225 TI - Cell-surface heparan sulfate is involved in the repulsive guidance activities of Slit2 protein. AB - Slit proteins are a family of secreted guidance proteins that can repel neuronal migration and axon growth via interaction with their cellular roundabout receptors (Robos). Here it was shown that Slit2-Robo-1 interactions were enhanced by cell-surface heparan sulfate. Removal of heparan sulfate decreased the affinity of Slit for Robo by about threefold. In addition, removal of cell surface heparan sulfate by heparinase III abolished the chemorepulsive response to Slit2 normally shown by both the migrating neurons and growing axons. These results indicate essential roles for cell-surface heparan sulfate in the repulsive activities of Slit2. PMID- 11426226 TI - CXCR4-activated astrocyte glutamate release via TNFalpha: amplification by microglia triggers neurotoxicity. AB - Astrocytes actively participate in synaptic integration by releasing transmitter (glutamate) via a calcium-regulated, exocytosis-like process. Here we show that this process follows activation of the receptor CXCR4 by the chemokine stromal cell-derived factor 1 (SDF-1). An extraordinary feature of the ensuing signaling cascade is the rapid extracellular release of tumor necrosis factor-alpha (TNFalpha). Autocrine/paracrine TNFalpha-dependent signaling leading to prostaglandin (PG) formation not only controls glutamate release and astrocyte communication, but also causes their derangement when activated microglia cooperate to dramatically enhance release of the cytokine in response to CXCR4 stimulation. We demonstrate that altered glial communication has direct neuropathological consequences and that agents interfering with CXCR4-dependent astrocyte-microglia signaling prevent neuronal apoptosis induced by the HIV-1 coat glycoprotein, gp120IIIB. Our results identify a new pathway for glia-glia and glia-neuron communication that is relevant to both normal brain function and neurodegenerative diseases. PMID- 11426227 TI - Visualization of changes in presynaptic function during long-term synaptic plasticity. AB - Controversy exists regarding the site of modification of synaptic transmission during long-term plasticity in the mammalian hippocampus. Here we used a fluorescent marker of presynaptic activity, FM 1-43, to directly image changes in presynaptic function during both short-term and long-term forms of plasticity at presynaptic boutons of CA3-CA1 excitatory synapses in acute hippocampal slices. We demonstrated enhanced presynaptic function during long-term potentiation (LTP) induced either chemically (with tetraethylammonium), or by high-frequency (200 Hz) electrical stimulation. Both of these forms of LTP required activation of L type voltage-gated calcium channels and NMDA receptors in the postsynaptic CA1 neuron. These results thus implied that a long-lasting increase in the efficacy of synaptic transmission is likely to depend, at least in part, on enhanced transmitter release from the presynaptic neuron. PMID- 11426228 TI - Kainate receptor-dependent axonal depolarization and action potential initiation in interneurons. AB - Kainate receptor agonists are powerful chemoconvulsants and excitotoxins. These properties are in part explained by depolarization of hippocampal principal neurons. However, kainate also depresses evoked inhibitory signals in pyramidal neurons, and promotes spontaneous GABA release from interneurons. The mechanisms underlying these phenomena are not fully understood, nor are the consequences for the inhibitory traffic among interneurons. We report that both the amplitude and the frequency of spontaneous IPSCs recorded in interneurons were enhanced by low concentrations of kainate, but action potential-independent IPSCs were unaffected. In the presence of GABA(A) receptor antagonists, kainate lowered the threshold for antidromic action potential generation, suggesting that interneuron axons are directly depolarized; this effect was mimicked by synaptically released glutamate. Kainate application also induced spontaneous antidromic action potentials. Axonal receptors are thus important in initiating the intense interneuronal activity triggered by kainate, which in turn influences inhibitory signaling to principal cells. PMID- 11426229 TI - Two different lateral amygdala cell populations contribute to the initiation and storage of memory. AB - Single-cell activity was recorded in the dorsal subnucleus of the lateral amygdala (LAd) of freely behaving rats during Pavlovian fear conditioning, to determine the relationship between neuronal activity and behavioral learning. Neuronal responses elicited by the conditioned stimulus typically increased before behavioral fear was evident, supporting the hypothesis that neural changes in LAd account for the conditioning of behavior. Furthermore, two types of these rapidly modified cells were found. Some, located in the dorsal tip of LAd, exhibited short-latency responses (<20 ms) that were only transiently changed. A second class of cells, most commonly found in ventral regions of LAd, had longer latency responses, but maintained enhanced responding throughout training and even through extinction. These anatomically distinct cells in LAd may be differentially involved in the initiation of learning and long-term memory storage. PMID- 11426230 TI - A neural circuit for circadian regulation of arousal. AB - An unknown aspect of behavioral state regulation is how the circadian oscillator of the suprachiasmatic nucleus (SCN) regulates sleep and waking. In this report, we describe the necessary elements for a circuit that provides circadian regulation of arousal. Trans-synaptic retrograde tracing revealed a prominent indirect projection from the SCN to the noradrenergic nucleus locus coeruleus (LC), a brain arousal system. Double-labeling experiments revealed several possible links between the SCN and the LC, including the dorsomedial (DMH) and paraventricular hypothalamic nuclei (PVN), as well as medial and ventrolateral pre-optic areas. Lesion studies confirmed that the DMH is a substantial relay in this circuit. Next, neurophysiology experiments revealed circadian variations in LC impulse activity. Lesions of the DMH eliminated these circadian changes in LC activity, confirming the functionality of the SCN-DMH-LC circuit. These results reveal mechanisms for regulation of circadian and sleep-waking functions. PMID- 11426231 TI - Compulsory averaging of crowded orientation signals in human vision. AB - A shape can be more difficult to identify when other shapes are near it. For example, when several grating patches are viewed parafoveally, observers are unable to report the orientation of the central patch. This phenomenon, known as 'crowding,' has historically been confused with lateral masking, in which one stimulus attenuates signals generated by another stimulus. Here we show that despite their inability to report the orientation of an individual patch, observers can reliably estimate the average orientation, demonstrating that the local orientation signals are combined rather than lost. Our results imply that crowding is distinct from ordinary masking, and is perhaps related to texture perception. Under crowded conditions, the orientation signals in primary visual cortex are pooled before they reach consciousness. PMID- 11426232 TI - Form constraints in motion binding. AB - Visual analyses of form and motion proceed along parallel streams. Unified perception of moving forms requires interactions between these streams, although whether the interactions occur early or late in cortical processing remains unresolved. Using rotating outlined shapes sampled through apertures, we showed that binding local motions into global object motion depends strongly on spatial configuration. Identical local motion components are perceived coherently when they define closed configurations, but usually not when they define open configurations. Our experiments show this influence arises in early cortical levels and operates as a form-based veto of motion integration in the absence of closure. PMID- 11426233 TI - Cerebral mechanisms of word masking and unconscious repetition priming. AB - We used functional magnetic resonance imaging (fMRI) and event-related potentials (ERPs) to visualize the cerebral processing of unseen masked words. Within the areas associated with conscious reading, masked words activated left extrastriate, fusiform and precentral areas. Furthermore, masked words reduced the amount of activation evoked by a subsequent conscious presentation of the same word. In the left fusiform gyrus, this repetition suppression phenomenon was independent of whether the prime and target shared the same case, indicating that case-independent information about letter strings was extracted unconsciously. In comparison to an unmasked situation, however, the activation evoked by masked words was drastically reduced and was undetectable in prefrontal and parietal areas, correlating with participants' inability to report the masked words. PMID- 11426234 TI - Reversal of subjective temporal order due to arm crossing. AB - How does the brain order successive events? Here we studied whether temporal order of two stimuli delivered in rapid succession, one to each hand, is determined before or after the stimuli are localized in space. When their arms were crossed, subjects could accurately report the temporal order, even when the interval between stimuli was as short as 70 ms. In most trials, subjects could also judge temporal order when their arms were crossed, but only if given adequate time (>1 s). At moderately short intervals (<300 ms), crossing the arms caused misreporting (that is, inverting) of the temporal order. Thus, at these intervals, the determining factor of temporal order was the spatial location of the hands. We suggest that it is not until the spatial locations of the hands are taken into account that the cutaneous signals from the respective hands are ordered in time. PMID- 11426235 TI - Going beyond the curriculum to promote medical education and practice in Saudi Arabia. AB - The foundation of health starts as early as undergraduate medical education. Medical education in the Kingdom has always been successful in promoting the medical profession and population health. The current issue in the Kingdom is quality assurance in all organizations including the health field. Thus emerges the value of evaluating the efficiency and effectiveness of the current system in meeting health needs and expectations. The aim of this paper was to analyze the current situation in order to design a frame for the direction of promotion of medical education and practice that best meets health needs and expectations. In fact, medical practice is a multidisciplinary process that is showing continuously changing theories. It has a broad scope of serving the whole community and improving the quality of life of all population categories. The huge quantity of information, needed to be retained by medical professionals, necessitates the move away from traditional methods of education to more practical and comprehensive programs of study. Most of the recent reform in medical education, in the Kingdom, has focused on curriculum and disregarded the education process. The requisite for the education process is a complete model of community-based health care, education and research. This study proposes a design aiming to enhance medical education and promote the medical profession, through developing the quality of medical professionals, that will foster growth of their activity and productivity, moving them into the community where clinical practice and experience are more relevant to the true health situation. PMID- 11426236 TI - Evidence-based medicine. AB - Evidence-based medicine is one of the recently born explosively growing important issues, whose philosophical origins extend back to mid-19th century and remains a hot topic for clinicians, public health practioners, purchasers, planners, and the public. Evidence-based health care has extended the application of the prinicples of evidence-based medicine to all professions associated with health care, including purchasing and management. So it is important for the up and coming young doctors and clinicians, and even other health care professions, to gain knowledge of critical appraisal and experience in the practice of evidence based health care. This article gives a brief description of what evidence-based medicine is and how to practice evidence-based medicine and enlists some useful Internet sites that can provide assistance in understanding the subject in even greater detail. PMID- 11426237 TI - Management of Crigler-Najjar Syndrome type I. AB - Crigler-Najjar Syndrome type I is a rare congenital disease with high mortality and morbidity rates due to brain complications. It has been treated by life-long phototherapy until the era of liver transplantation. Liver transplantation is currently the only curative treatment for this syndrome. Liver transplantation prevents the severe neurological complications that are the main cause of life long disability in Crigler-Najjar Syndrome type I. The ideal age for transplantation is 3-5 years. Despite the advent of auxiliary transplantation we believe that orthotopic liver transplantation is the optimal treatment and the ideal method of liver transplantation for Crigler-Najjar Syndrome type I. PMID- 11426238 TI - Iron deficiency anemia. A study of risk factors. AB - OBJECTIVE: To determine the risk factors for iron deficiency anemia among Saudi women of childbearing age. METHODS: This is a case control study conducted at the primary health care clinics of King Khalid University Hospital in Riyadh over a 6 month period. Inclusion criteria included women of childbearing age and hemoglobin level < 120 g/l. Iron deficiency anemia was defined as having iron deficiency and low hemoglobin level < 120 g/l. Controls had hemoglobin level > 120 g/l and were matched with the cases for socio-demographic characteristics. RESULTS: Eighty-seven patients and 203 controls were enrolled in the study. Low frequency of eating meat, vegetables or drinking juices right with vitamin C increased the risk of having iron deficiency anemia by 2-4 fold (odds ratio = 2.06, 95% confidence interval 1.20-3.54), (odds ratio = 2.86, 95% confidence interval 1.65-4.98) and (odds ratio = 3.75, confidence interval 2.20 - 6.42). Menstrual period duration of > 8 days, history of clots or flooding increased the odds of having iron deficiency anemia by 3-6 fold. The odds of being iron deficient in patients on non-steroidal anti-inflammatory drugs and antacid were 6 9 fold. CONCLUSION: Important risk factors for iron deficiency anemia among Saudi women of childbearing age are dietary habits, menorrhagia and history of ingestion of non-steroidal anti-inflammatory drugs or antacids. PMID- 11426239 TI - Differentiated thyroid carcinoma referred for radioiodine therapy. AB - OBJECTIVE: The current work was conducted to study the disease status and treatment results of patients with differentiated thyroid carcinoma referred for radioactive iodine therapy. METHODS: Retrospective review of 78 patients with differentiated thyroid carcinoma referred for radioiodine therapy in the Nuclear Medicine Unit, King Abdulaziz Hospital and Oncology Center, Jeddah, Kingdom of Saudi Arabia. Analysis of the clinicopathologic characteristics, age correlation to different risk factors, treatment protocol and results were performed. RESULTS: Seventy seven percent were female and the female to male ratio was 3.5:1. The age of patients ranged between 13-63 years with a median age of 36 years. Cervical lymph node involvement was detected in 22 patients (25%). Papillary carcinoma was encountered in 78 patients (90%) and follicular carcinoma in 9 patients (10%). Analysis of the clinicopathologic characteristics showed no statistically significant difference between patients in the different age groups except for extrathyroid extension and lymph node involvement. Patients older than 45 years had a statistically significant lower incidence of nodal involvement and higher incidence of extra thyroid extension (P<0.02). In the current study we used a high dose method (Radioiodine-131 dose 75-100mCi) for thyroid remnant ablation after thyroidectomy (total or near total) in 67 patients. An Iodine 131 dose of 150 mCi was used in 12 patients with radioiodine-avid cervical lymph nodes and in 3 patients with gross residual tumor. In 4 patients with distant metastases an Iodine 131 dose of 200 mCi was used. For the whole study group the 5 year overall survival and disease-free survival was 96% and 88%. CONCLUSION: The current study, as with many other retrospective studies, concluded that despite the fact that differentiated thyroid carcinoma is among the most curable cancers, some patients are still at high risk for recurrent disease and associated mortality. PMID- 11426240 TI - The performance profile of medical students in the mock objective structured clinical examination. AB - OBJECTIVE: Acquisition of clinical skills, after completing a course in the basic sciences, is an essential aspect of undergraduate training in any medical school. These skills are usually divided into 3 broad categories: namely, history taking, physical examination and selection of the appropriate laboratory investigations. At the end of the clerkship, the students' clinical skills have to be assessed. The main objective was to describe the performance profile of a group of medical students while examing a distended abdomen in the United Arab Emirates. METHODS: This paper describes the performance profile of 24 randomly selected medical students in a mock objective structured clinical examination of a patient with distended abdomen in the hospital environment. Marks were allotted according to the checklist of the performance expectations. For the assessment used in this paper, ability to perform a step of the clinical examination was rated positive and documented. RESULTS: The performance profile of the students was very good to excellent, corresponding to 9 out of 10 marks. All students identified or excluded the common signs of clinical ascites. The signs uncommonly seen in this area, such as Dupytren's contractures, were excluded by 20 of 24 (83%) students. CONCLUSION: The excellent performance is attributed to a greater exposure to patients with mainly gastrointestinal disorders during the clerkship. The main advantage of an objective structured clinical abdominal examination, which is set up in a hospital environment, is that it reflects the real life situation of a practising physician, as opposed to using simulated patients. Although, a structured clinical examination is labor-intensive and costly, the advantages outweigh the work and cost of setting it up. PMID- 11426241 TI - Hope for clomiphene citrate resistant ovaries. AB - OBJECTIVE: To assess the presentation, different surgical modalities and their outcome in the treatment of polycystic ovarian disease. METHODS: A retrospective study was carried out on 66 patients with polycystic ovarian disease. The majority of the cases had a history of infertility for at least 2 years and were known to be clomiphene citrate resistant in the Saudi German Hospital, Jeddah between 1994 and 1998 undergoing laparoscopic ovarian drilling. RESULTS: More than 80% of the patients were below 30 years of age. The majority of cases below 30 years had primary infertility. Irregular cycle was the main clinical presentation in 75% of cases; ovarian volume was more than 5.5cm3 in all of the cases. Diathermy drilling was carried out for 50 patients, laser for 12 patients and wedge resection for 4 patients. The follow-up treatment showed that pregnancy rate was 23%, 38% and 0% in the 3 groups. Cases that failed to conceive had significantly higher luteinizing hormone level. CONCLUSION: Laparoscopic ovarian drilling using diathermy or laser is effective during the induction of ovulation and increases the pregnancy rate in cases of polycystic ovarian disease resistant to clomiphene citrate. PMID- 11426242 TI - Contraceptive use in Jordan. AB - OBJECTIVE: To compare women in the child bearing age for the years 1983, 1990, 1997 regarding their use of contraceptives and the variations in the source of intrauterine contraceptive devices, as it was recently observed that women are more likely to use intrauterine contraceptive devices. METHODS: Three national surveys carried out in 1983, 1990, 1997 were compared. The samples selected in the 3 surveys were women in the child bearing age. A multistage sampling procedure was used to ensure the representativeness of the samples in the 3 surveys. RESULTS: The use of contraceptives increased from 26% in 1983 to 40% in 1990 and 53% in 1997. Women became more likely to use intrauterine contraceptive devices where the percentage of use increased from 32% in 1983 to 44% in 1997. The percentage of users of contraceptives in general and intrauterine contraceptive devices in particular was higher in urban areas and in educated women with a high number of children. CONCLUSION: Women at risk of not using contraceptives should be identified and contacted to convince them to use contraceptives. All barriers to access, use and quality of contraceptives should be investigated. The identification of such barriers will be useful for decision makers and health planners in the country. PMID- 11426243 TI - Visceral Leishmaniasis in children in the Yemen. AB - OBJECTIVE: The clinical presentation and duration of therapy for visceral leishmaniasis varies in different countries. The sodium stibogluconate is costly, and a trial of short course therapy has not yet been studied in Hajjah governorate. The aim of this study was to evaluate the efficacy of a 20 days regimen of sodium stibogluconate and to ascertain the epidemiological, clinical and laboratory features of visceral leishmaniasis in children. METHODS: This was a prospective hospital-based study in Hajjah Governorate, Republic of Yemen. Children of 12 years of age or less with a confirmed diagnosis were included. Sodium stibogluconate was given in a dose of 15mg/kg/dose daily for 20 days, then the patients were re-evaluated and the data required for achieving the other objective was collected. RESULTS: Thirty-two patients fulfilled the inclusion criteria. The age ranged from 12 months to 144 months (67.7 +/- 35). Females formed 53% of this criteria. The duration of symptoms ranged from 2 weeks to 116 weeks. Fever, fatigability and abdominal distension were the most common symptoms. The hematological findings showed anemia in all patients, leukopenia in 81% and thrombocytopenia in 56%. Formol gel test was negative in 20 patients (63%). Malaria smear was positive in 11 patients (34%). Splenic aspiration was carried out in 25 patients (78%) and bone marrow aspiration in 7 patients (22%). Blood transfusion were required for 24 patients (73%). After 20 days treatment with pentostam, 20 patients (63%) came for follow-up and re-tested for parasitological cure. Half of those were still positive for leishmania donovan bodies. The mortality rate was 5%. CONCLUSION: The clinical features were of the Mediterranean type. Twenty days treatment with sodium stibogluconate was not adequate. PMID- 11426244 TI - Clinicopathological patterns and distribution of Schistosomiasis in Asir Region. AB - OBJECTIVE: The objective of this study is to report, for the first time, the histopathologic pattern of Schistosomiasis from the Asir Region and to compare it to patterns reported from other regions of the Kingdom of Saudi Arabia. Several studies have reported the patterns of Schistosomiasis in other regions of the Kingdom of Saudi Arabia and other countries where Schistosomiasis is endemic. Schistosomiasis is endemic in certain areas of Asir region, however no data is available concerning the clinicopathological pattern of Schistosomiasis in the Asir Region. METHODS: This is a retrospective analysis of 217 cases of Schistosomiasis from surgical and biopsy files of Asir Central Hospital during a period from January 1990 to October 1999. RESULTS: Our study revealed that Schistosomiasis was more common among the expatriate population of Asir Region than Saudi nationals residing in this area. The urinary tract was most commonly involved, and then in descending frequency came the vermiform appendix, liver and large bowel. CONCLUSION: These findings are somewhat different from those observed in the Riyadh Region where the vermiform appendix was the most commonly affected organ. Based on the histopathologic pattern, our study describes the pattern of Schistosomiasis in the Asir Region and may serve as a base-line for future research work. PMID- 11426245 TI - Accidental injuries during muslim pilgrimage. AB - OBJECTIVE: The diseases that occur during Hajj are a unique medical problem. The aim of this study is to highlight the incidence, the type and the early surgical management of trauma during the Hajj period. METHODS: A prospective study was carried out during a 15 day-period conducted in two parts in the emergency room, and a follow-up of the admitted patients in the surgical department. RESULTS: The number of trauma cases who attended the emergency department was 713 patients, 248 patients were admitted in different surgical departments, together with the intensive-care unit. Sixty-five percent of the patients were discharged from emergency room as they had minor trauma. Sixty percent were involved in road traffic accidents, 15% had injuries in Holy Haram. Limb fracture accounts for 53% of total trauma admissions. Two patients died in the first 48 hours. CONCLUSION: Trauma during Hajj is a real surgical problem which deserves more attention. Orthopedic and Neurosurgical cases are the most common surgical cases during the Hajj period. PMID- 11426246 TI - Regulation of polymorphonuclear leukocyte function by platelets. AB - OBJECTIVE: To investigate the full effect of platelet-derived constituents on various polymorphonuclear neutrophil leukocyte responses. METHODS: Polymorphonuclear neutrophil leukocytes and platelets were separated from fresh blood of normal healthy volunteers. Platelets were then stimulated partially, or maximally to release constituents of their a- or a- and dense granules. The effects of these constituents on polymorphonuclear neutrophil leukocyte function (oxidase activity, degranulation and migration) were investigated. RESULTS: Platelet-derived constituents were found to both enhance, and inhibit polymorphonuclear neutrophil leukocytes-oxidant production, depending on the incubation time. Enhancement was due to dense granule-derived nucleotides (adenosine diphosphate and adenosine diphosphate), while inhibition was due to adenosine monophosphate derived from these nucleotides by polymorphonuclear neutrophil leukocyte surface nucleotidases. This latter inhibitory effect was reversed by the cytokine, granulocyte-macrophage colony stimulating-factor. Moreover, platelet constituents consistently enhanced other polymorphonuclear neutrophil leukocyte responses including degranulation and migration regardless of the incubation period. The latter enhancement was due to a-granule constituents, most likely platelet factor 4. CONCLUSION: Platelets, through release of their granular constituents, are able to modulate polymorphonuclear neutrophil leukocyte function in a way that is physiologically beneficial. PMID- 11426247 TI - Evaluation of parental knowledge of Pediatric Rheumatic Diseases. AB - OBJECTIVE: To investigate the parental knowledge of pediatric rheumatic diseases in general, and in particular information regarding their children's diseases. To focus on the important role of health education in understanding these chronic diseases, and formulate a future plan for establishing a general public education program. METHODS: One hundred sets of parents, of children with established rheumatic diseases with mean duration of illness, (4.1 +/- 2.83), a mean child age (9.9 +/- 3.15) years, were given a 20 multiple choice questionnaire during their routine visit to the Pediatric Rheumatology Clinic and Physiotherapy Department or to the pediatric ward at King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia, over a 6-month period between December 1998 and June 1999. The questionnaire addressed 4 main areas: 1. parental awareness regarding their children's diagnosis and duration, 2. source of information and parental satisfaction, 3. general knowledge about rheumatic diseases and 4. knowledge about medical and physical therapy. The total possible score is 23. RESULTS: The overall mean score is 11.6. The mean total score is not affected by the parental education level. The majority of parents have wrong beliefs, regarding rheumatic diseases. The treating physician is the main source of parental information and in the majority of the parents, this information is satisfactory. CONCLUSION: The questionnaire is a simple and easy test to investigate parental knowledge regarding pediatric rheumatic diseases. The survey shows the need for health education programs and a future general public health education plan to improve awareness of pediatric rheumatic diseases and maybe other chronic illness. PMID- 11426248 TI - Urinary tract infections in general practice in a rural community in South Trinidad. AB - OBJECTIVE: A prospective study was carried out at a General Practice in South Trinidad, to assess the sensitivity profiles of urinary isolates in this remote rural community. METHODS: Quantitative bacteriologic cultures were performed according to standard procedures. Identification of isolates was based on gram reaction, morphology and biochemical reactions. Susceptibility testing was carried out using commonly prescribed antimicrobials for the treatment of urinary tract infections. RESULTS: From 779 urine specimens, 49% were culture positive for counts >105 organisms per ml. Three hundred and eighteen were aged 21 years and above and 85.5% of these were females. The lowest incidence of urinary tract infections, 8%, was seen among the 13-20 year old age group. Most males above 50 years had indwelling urethral catheters due to prostatic disease. Escherichia coli was the predominant isolate, 71%, followed by Proteus mirabilis, 9%, and Klebsiella pneumoniae, 7%. All urinary tract infections isolates were fully sensitive to ofloxacin and >96% (except for Citrobacter species, 63%) were sensitive to cefuroxime. Apart from Enterococcus faecalis and Staphylococcus saprophyticus, the overall sensitivity to ampicillin by all isolates was <33%. The prevalence of multi-resistant Pseudomonas aeruginosa in community-acquired urinary tract infections is increasing. All Pseudomonas aeruginosa were fully sensitive to gentamicin and ofloxacin. CONCLUSION: The high frequency of single and multiple antibiotic resistances in this rural community should cause alarm. The ease of procuring antibiotics in these areas without a prescription can result in uncontrolled self-medication. Also, the absence of antibiotic prescribing policies and inadequate information on patterns of bacterial resistance, may all contribute to the emergence of resistant strains. PMID- 11426249 TI - Lymphomatoid granulomatosis with long survival. AB - We present a case of lymphomatoid granulomatosis presenting in a 35-year-old Saudi lady with long survival. She responded to treatment with intermittent cyclophosphamide infusion in addition to corticosteroids. This is the first case of lymphomatoid granulomatosis to be reported in the Arab world. The prolonged survival and response to intermittent cyclophosphamide infusion is discussed. PMID- 11426250 TI - Caudal Regression Syndrome and peno-scrotal transposition. AB - Caudal Regression Syndrome is a rare anomaly affecting different systems in the caudal portion of the fetus. Peno-scrotal transposition is a rare condition of variable severity often associated with Caudal Regression Syndrome. We report a case of Caudal Regression Syndrome with perineal ano-rectal duplication, incomplete peno-scrotal transposition, perineal hypospadias, chordee, and thoracic hemivertebrae. PMID- 11426251 TI - The role of radiological and gravity axis parameters in the assessment of the arch of the foot. PMID- 11426252 TI - Frozen section experience. PMID- 11426253 TI - Emerging diseases, local systems, and globalization. PMID- 11426254 TI - An ecosystem approach to health and its applications to tropical and emerging diseases. AB - Disease and health outcomes occur within a complex socio-ecological context characterized by feedback loops across space and time, self-organization, holarchies, and sudden changes in organization when thresholds are reached. Disease control programs, even if they are successful, may undermine health; conversely, programs in agriculture and economic development designed to improve health may simply alter disease patterns. A research and development strategy to promote sustainable health must therefore incorporate multiple scales, multiple perspectives, and high degrees of uncertainty. The ecosystem approach developed by researchers in the Great Lakes Basin meets these criteria. This has implications for community involvement in research, development policies, and for understanding and controlling tropical and emerging diseases. Even if unsuccessful in achieving specific outcome targets, however, the requirements of this approach for open and democratic communication, negotiation, and ecological awareness make its implementation worthwhile. PMID- 11426264 TI - Human health improvement in Sub-Saharan Africa through integrated management of arthropod transmitted diseases and natural resources. AB - A concept of an ecosystem approach to human health improvement in Sub-Saharan Africa is presented here. Three factors mainly affect the physical condition of the human body: the abiotic environment, vector-transmitted diseases, and natural resources. Our concept relies on ecological principles embedded in a social context and identifies three sets of subsystems for study and management: human disease subsystems, natural resource subsystems, and decision-support subsystems. To control human diseases and to secure food from resource subsystems including livestock or crops, integrated preventive approaches are preferred over exclusively curative and sectorial approaches. Environmental sustainability - the basis for managing matter and water flows - contributes to a healthy human environment and constitutes the basis for social sustainability. For planning and implementation of the human health improvement scheme, participatory decision support subsystems adapted to the local conditions need to be designed through institutional arrangements. The applicability of this scheme is demonstrated in urban and rural Ethiopia. PMID- 11426265 TI - [Regional differences and Hantavirus pulmonary syndrome]. AB - Factors related to the characteristics of Hantavirus pulmonary syndrome in Argentina are described. Factors from different scientific fields converge to form the syndrome's analytical framework. Some of these factors are the history of spatial occupation, work and production structures, human migration patterns, ethnic composition, reservoir dynamics and its relationship to the different circulating viruses, and human behavior. Furthermore, the multiple factors are expressed in three ecological frameworks, associated with three different geographical regions of Argentina: 1) Northwest; 2) Central ("wet Pampa"); and 3) South Andean. In order to understand the actual causality of health or disease as an interaction of many factors, research on the primary biological, social, and environmental determinants of diseases should attend to the complexity of variable relationships in each region. The multiple-factor convergence approach allows for the design of appropriate strategies to improve the population's health status. Therefore, strategies should be developed and transferred by multidisciplinary teams, while their sustainability should be assured by community participation beginning with the earliest steps of research onward. PMID- 11426266 TI - The place behind the case: leptospirosis risks and associated environmental conditions in a flood-related outbreak in Rio de Janeiro. AB - The environmental context in which a leptospirosis outbreak took place during the summer of 1996 in the Rio de Janeiro Western Region was examined by using spatial analysis of leptospirosis cases merged with socio-demographic data using Geographic Information System (GIS). Risk areas were mapped based on flood and solid waste accumulation information for the region. Incidence rates were calculated for each area by the division of number of cases per total population in the specific areas. Higher rates were observed for census tracts inside the flood risk area and in the vicinities of waste accumulation sites. These findings are in agreement with the expected risk of leptospirosis, evidencing the role of environmental and collective factors in the determination of the disease. PMID- 11426267 TI - Ecosystem approaches to human health. AB - The promotion of human health must be embedded in the wider pursuit of ecosystem health. Interventions will be impaired if ecosystem-linked determinants of health are not taken into account. In the extreme case, if ecosystems lose their capacity for renewal, society will lose life support services. Essential features of ecosystem health are the capacity to maintain integrity and to achieve reasonable and sustainable human goals. An ecosystem approach to research and management must be transdisciplinary and assure participation of stakeholders. These requisites provide a means for science to better deal with the complexity of ecosystems, and for policy-makers and managers to establish and achieve reasonable societal goals. The ecosystem approach can determine links between human health and activities or events which disturb ecosystem state and function. Examples are: landscape disturbance in agriculture, mining, forestry, urbanization, and natural disasters. An understanding of these links can provide guidance for management interventions and policy options that promote human health. An ecosystem approach to management must be adaptive because of irreducible uncertainty in ecosystem function. PMID- 11426268 TI - [Living conditions and life experiences of working-class groups in Rio de Janeiro: rethinking dengue control and popular mobilization]. AB - Using narratives of an experience with popular mobilization during the 1986-91 dengue epidemic in the city of Rio de Janeiro, the authors discuss the scientific research and technical counseling involving basic sanitation conditions for vulnerable social groups. They present research results on water distribution in the slums from the Leopoldina area of the city. The research stemmed from demands by community leaders at local forums discussing health conditions. Gathering, systematizing, and analyzing the data were based on what they call "shared knowledge construction", resulting by crossing accumulated scientific knowledge with popular knowledge produced as a result of living conditions and life experiences among working-class groups. Finally, the authors comment on the need for local health professionals to be aware of relationships between epidemic and endemic processes and protection of life. PMID- 11426269 TI - Chagas disease prevention through improved housing using an ecosystem approach to health. AB - This Chagas disease prevention project via housing improvement aims to determine the efficiency of different interventions in vector control. The following study describes the target communities, disease magnitude, and housing improvements. Transmission levels are analysed from an ecological and socioeconomic perspective. Special interest was focused on the peridomicile as the origin of domiciliary reinfestation. In the original project, three intervention programs were proposed, one for each of the three communities: (a) an insecticide spraying program; (b) a housing improvement program; and (c) a combined program of spraying and housing improvement. The three communities currently have different risks of exposure to triatominae reinfestation as a consequence of the type of intervention carried out. A new multidisciplinary approach which integrates participatory, community-based research and socioeconomic dimensions will allow to determine the efficiency of models for territorial ordering, community education, and environmental interventions in Chagas disease control. PMID- 11426270 TI - [Urbanization and dengue ecology]. AB - Demographic changes occurring in underdeveloped countries due to intense rural urban migration since the 1960s have resulted in overcrowded cities with multiple deficiencies, particularly in housing and basic sanitation. Some 20% of the population in large and medium-sized cities live in slums or under similar conditions. Lack of regular water supply and public garbage collection foster the proliferation of potential breeding sites for Aedes aegypti (the main mosquito vector for dengue), including precarious reservoirs for potable water and disposable recipients which accumulate water, like used cans and plastic and glass bottles. Modern industries also produce large volumes of disposable materials. Propagation of the dengue virus and the spread of dengue vectors are favored by the high intensity, frequency, and speed of private and public transportation. Such factors can help explain the re-emergence of dengue, the most important arbovirus in the world today, affecting thousands of people each year. PMID- 11426271 TI - An integrated malaria control program with community participation on the Pacific Coast of Colombia. AB - The study focuses on integrated malaria control in 23 communities on the Pacific Coast of Colombia, with several elements of an ecosystem approach to human health, including malaria-related sociopolitical, ecological, and economic factors. The program fostered community participation. The program presented here had 2 components: implementation and research. The first was conducted in 23 communities, 21 of which lacked adequate health services in terms of education, community participation, prompt diagnosis and complete treatment, and vector control. Research focused on specific vector control measures and the current national health services decentralization process. The project: 1) created a malaria prevention culture in the community; 2) avoided deaths from malaria (no fatal cases in the 3-year period, compared to 5-8 deaths a year previously); 3) avoided cases of cerebral malaria (no cases, as compared to 90-110 per year previously); 4) reduced malaria incidence by 45.36%; 5) decreased length of sick leave from 7.52 to 3.7 days; 6) established a permanent network of microscope technicians and 2-way radio communications; 7) integrated work by local, regional, and outside institutions; 8) demonstrated efficacy of insecticide impregnated bednets to reduce malaria transmission. PMID- 11426272 TI - Interactions between global processes and local health problems. A human ecology approach to health among indigenous groups in the Amazon. AB - This article deals with methodological issues and how to link global processes - social and ecological - with environmental changes and human health in local communities. The discussion concerns how interdisciplinary approaches can help us find tools to develop new knowledge. Scientific knowledge and local knowledge are not seen as opposite epistemological forms, but as socially and culturally constructed. Power and social legitimacy have to be included when analyzing how to deal with the interaction between global processes and local environmental change and the health/disease interface. PMID- 11426273 TI - Wetlands and infectious diseases. AB - There is a historical association between wetlands and infectious disease that has led to the modification of wetlands to prevent disease. At the same time there has been the development of water resources projects that increase the risk of disease. The demand for more water development projects and the increased pressure to make natural wetlands economically beneficial creates the need for an ecological approach to wetland management and health assessment. The environmental and health interactions are many. There is a need to take into account the landscape, spatial boundaries, and cross-boundary interactions in water development projects as well as alternative methods to provide water for human needs. The research challenges that need to be addressed are discussed. PMID- 11426274 TI - Climate-disease connections: Rift Valley Fever in Kenya. AB - All known Rift Valley fever(RVF) outbreaks in Kenya from 1950 to 1998 followed periods of abnormally high rainfall. On an interannual scale, periods of above normal rainfall in East Africa are associated with the warm phase of the El Nino/Southern Oscillation (ENSO) phenomenon. Anomalous rainfall floods mosquito breeding habitats called dambos, which contain transovarially infected mosquito eggs. The eggs hatch Aedes mosquitoes that transmit the RVF virus preferentially to livestock and to humans as well. Analysis of historical data on RVF outbreaks and indicators of ENSO (including Pacific and Indian Ocean sea surface temperatures and the Southern Oscillation Index) indicates that more than three quarters of the RVF outbreaks have occurred during warm ENSO event periods. Mapping of ecological conditions using satellite normalized difference vegetation index (NDVI) data show that areas where outbreaks have occurred during the satellite recording period (1981-1998) show anomalous positive departures in vegetation greenness, an indicator of above-normal precipitation. This is particularly observed in arid areas of East Africa, which are predominantly impacted by this disease. These results indicate a close association between interannual climate variability and RVF outbreaks in Kenya. PMID- 11426275 TI - Public health challenges and emerging diseases: the case of Sao Paulo. AB - The author discusses the challenges posed by emerging infectious diseases in 100 years of public health in the state of Sao Paulo, Brazil. With an advanced and organized public health system, Sao Paulo responded to the emergence of infectious diseases by creating research institutions and control programs. The late 19th century witnessed the first modern research institution in microbiology, in response to the bubonic plague. A changing economy led to constant changes in ecosystems. The late 20th century presents a wide array of both emerging and rapidly changing infectious diseases. The present situation calls for creative solutions. Ecosystem analysis and more agile epidemiological surveillance are seen as the best alternatives. PMID- 11426276 TI - Global surveillance of emerging diseases: the ProMED-mail perspective. AB - The Internet is changing the way global disease surveillance is conducted. Countries and international organizations are increasingly placing their outbreak reports on the Internet, which speeds up distribution and therefore prevention and control. The World Health Organization (WHO) has recognized the value of nongovernmental organizations and the media in reporting outbreaks, which it then attempts to verify through its country offices. However, WHO and other official sources are constrained in their reporting by the need for bureaucratic clearance. ProMED-mail has no such constraints, and posts outbreak reports 7 days a week. It is moderated by infectious disease specialists who add relevant comments. Thus, ProMED-mail complements official sources and provides early warning of outbreaks. Its network is more than 20,000 people in over 150 countries, who place their computers and time at the network's disposal and report on outbreaks of which they have knowledge. Regions and countries could benefit from adopting the ProMED-mail approach to complement their own disease surveillance systems. PMID- 11426277 TI - Inadequate management of natural ecosystem in the Brazilian Amazon region results in the emergence and reemergence of arboviruses. AB - A total of 187 different species of arboviruses and other viruses in vertebrates were identified at the Evandro Chagas Institute (IEC) from 1954 to 1998, among more than 10,000 arbovirus strains isolated from humans, hematophagous insects, and wild and sentinel vertebrates. Despite intensive studies in the Brazilian Amazon region, especially in Para State, very little is known about most of these viruses, except for information on date, time, source, and method of isolation, as well as their capacity to infect laboratory animals. This paper reviews ecological and epidemiological data and analyzes the impact of vector and host population changes on various viruses as a result of profound changes in the natural environment. Deforestation, mining, dam and highway construction, human colonization, and urbanization were the main manmade environmental changes associated with the emergence and/or reemergence of relevant arboviruses, including some known pathogens for humans. PMID- 11426278 TI - [Chagas disease, environment, participation, and the state]. AB - Human Chagas Disease (HCD) affects 16 million Latin Americans and is related to environmental and social questions like education, anthropogenic changes, housing, and migration. HCD requires domiciliation of the vector, which is introduced from sylvatic ecotopes or transported passively by humans. HCD is difficult to treat but possible to prevent. As a disease concentrating among poor individuals and regions, its control requires government intervention, involving education, community participation, and access to medical care. Public policies are required for HCD control, requiring participation by political parties and social control of public agencies to ensure program continuity. A consistent environmental policy is needed to prevent the spread of HCD to new areas such as the Amazon Region. Finally, partnerships among affected countries can help foster HCD control, raising self-esteem and helping promote a new political order in Latin America. PMID- 11426279 TI - An ecosystem approach to malaria control in an urban setting. AB - We conducted a research project aimed at strengthening local government and the community for a sustainable malaria control strategy. The project began with a baseline diagnosis of malaria prevalence, a KAP survey, entomology, and health services delivery, after which an epidemiological study was performed to identify risk factors associated with malaria, thereafter used to plan intervention measures. A program evaluation was conducted five years later. By using an ecosystem approach to reanalyze data, this paper discusses how malaria arises from a complex interaction of cultural, economic, ecological, social, and individual factors. Intervention measures require an intersectorial and transdisciplinary approach that does not exist at the moment. Health sector leadership is limited, and there is no true community participation. Implications for research, including the use of qualitative and quantitative methods, study design, and complexity of data analysis are discussed. Finally, implications for malaria control are discussed, stressing the differences between the ecosystem and integrated disease control approaches. PMID- 11426280 TI - Health, biodiversity, and natural resource use on the Amazon frontier: an ecosystem approach. AB - This study aims to improve the health of rural Amazonian communities through the development and application of a participatory ecosystem approach to human health assessment. In the study area marked seasonal fluctuations dictate food availability, water quality and disease outbreak. Determining the causal linkages between ecosystem variables, resource use and health required a variety of forms of inquiry at multiple scales with local participation. Landscape spatial mapping of resource use demonstrated the diversity of the ecological resources upon which communities depend. Household surveys detailed family and individual consumption and production patterns. Anthropometric measurements, parasite loading, water quality and anemia levels were used as indicators of health status. This was complemented with an ethnographic and participatory health assessment that provided the foundation for developing community action plans addressing health issues. Discussion is focused on three attributes of an ecosystem approach; (a) methodological pluralism, (b) cross-scale interactions and (c) participatory action research. PMID- 11426281 TI - An ecosystem approach to human health and the prevention of cutaneous leishmaniasis in Tumaco, Colombia. AB - A study was conducted during 1996-1997 in 20 villages of Tumaco, Colombia, to evaluate the effectiveness of personal protective measures against cutaneous leishmaniasis (CL). The intervention was effective, but the high costs of the preventive measures and the lack of a more holistic approach hampered the intervention's sustainability. This paper analyzes the results using an ecosystem approach to human health. Using this approach, we found that CL has been present in the study area for a long time and affects farmers and those living closest to the forest. The forest constitutes the habitat for insect vectors (sandflies) and parasite reservoirs (wild mammals). Four spatial scales were identified in this ecosystem: residential, village, regional, and global. From the ecosystem perspective, three interventions are proposed to prevent CL in the 20 villages: improve housing construction, organize village housing in clusters, and make diagnosis and treatment of CL more accessible. The design and implementation of these interventions require active involvement by people with the disease (village inhabitants) and decision-makers (local authorities). PMID- 11426282 TI - Ecological approaches to rural development projects. AB - Most rural development projects include ecological considerations, and most conservation projects include some reference to sustainable development. However, conservation projects frequently fail because they do not incorporate local communities' perceptions and needs. Many development projects are also unsuccessful because they are not based on adequate ecological assessment. We focus here on the most important ecological issues to be addressed in order to place development projects in an ecosystem context. Such projects should incorporate updated and precise ecological concepts and methods. Some key ecological issues in development projects are the relationships between ecosystem functions, services, and sustainability, the concept of loose connectivity, the distinct and complementary concepts of ecosystem resistance and resilience, and the links between biodiversity and ecosystem functioning. We claim that an ecologically sound development project maximizes the preservation and improvement of ecosystem services, especially for local communities. We pose a series of questions aimed at placing rural development projects in an ecosystem context and suggest ways of organizing this information. PMID- 11426283 TI - [Emerging and reemerging viral diseases]. AB - Recent decades have witnessed previously unknown viruses like HIV, along with other previously controlled viruses like dengue. The most important mechanisms have been the emergence of new viral strains by genetic alterations, the breakdown of species barriers by viruses, and viral spread from ecological niches. The main factors facilitating such mechanisms have been demographic pressure, with the expansion of the agricultural frontier, social behavior patterns, intensive air traffic, transporting both vectors and infected humans, importation of animals carrying the viruses, large-scale ecological alterations like dam- and road-building, and the widespread transformation of health systems, with a reduction in resources and infrastructure for disease control activities. Discussions on an international scale have recommended investments in the areas of Epidemiological Surveillance, Research Applied to Public Health, an emphasis on disease prevention and vector control measures, and infrastructure improvements in the health sector at the local, State, and federal level to reduce the impact of these viral diseases. PMID- 11426284 TI - Active living for active aging: measuring, understanding and influencing physical activity in populations. PMID- 11426285 TI - Does aging change nutrition requirements? AB - The current adult guidelines for a healthy diet make no distinctions between adults aged 25-50 y and those aged 51 y and over. The question is whether dietary guidelines ought to be stratified by age, in recognition of the dietary and nutrient needs of the growing population of elderly adults. There are limited data on nutrient requirements of older adults. Aging is accompanied by a variety of physiological, psychological, economic and social changes that may adversely affect nutritional status. Older people have a higher prevalence of chronic disease, take multiple medications and supplements, and tend to be sedentary. Higher prevalence of obesity is difficult to reconcile with sharply lowered energy intakes. While basal metabolic rate does decline with age, lack of physical activity among the elderly is the more likely answer. The USDA Food Guide Pyramid is the key tool for communicating dietary advice to consumers. Researchers at Tufts University have produced a 70+ pyramid to reflect the nutrient needs of elderly adults. An additional focus has been on quality of life issues. Increased longevity is not always associated with continued good health. The Healthy People 2010 Objectives now use quality of life as a national health standard. Whereas health related quality of life (HRQOL) measures assess physical and mental health and their determinants, global quality of life measures focus on life satisfaction. Optimal nutrition promotes both functional health status and mental well-being. Dietary diversity and variety promotes enjoyment and satisfaction with the diet. Regular physical activity promotes strength and endurance, helps to maintain appropriate body weight, and contributes to independent physical functioning. Improving health-related quality of life is a key element in promoting the health and well-being of older adults. PMID- 11426286 TI - Impact of aging on eating behaviors, food choices, nutrition, and health status. AB - People eat less and make different food choices as they get older. It is unclear what impact these dietary changes may have on health status. However, lower food intake among the elderly has been associated with lower intakes of calcium, iron, zinc, B vitamins and vitamin E. Low energy intakes or low nutrient density of the diet may increase the risk of diet-related illnesses and so pose a health problem. Several factors may influence this observed decline in energy intake. Older adults tend to consume less energy-dense sweets and fast foods, and consume more energy-dilute grains, vegetables and fruits. Daily volume of foods and beverages also declines as a function of age. Physiological changes associated with age, including slower gastric emptying, altered hormonal responses, decreased basal metabolic rate, and altered taste and smell may also contribute to lowered energy intake. Other factors such as marital status, income, education, socioeconomic status, diet-related attitudes and beliefs, and convenience likely play a role as well. Many age-related nutritional problems may be remedied to some extent by providing nutrient-dense meals through home delivery or meal congregate programs. Management of medical and dental problems and the provision of vitamin and mineral supplements may also be effective. More studies that integrate nutrition research, public health intervention, and outcomes research are needed to determine the impact of diet on nutrition, health, and overall quality of life. PMID- 11426287 TI - Optimizing immunity and gut function in the elderly. AB - The elderly represent a group for whom health issues are of paramount concern. Physiological processes, including the functioning of the immune and digestive systems, are known to decline with age, and consequently aging can increase susceptibility to infectious and non-infectious disease. Dietary supplementation may offer a safe and effective means of countering physiological senescence, by optimizing important immune and gut functions. This review outlines our current understanding of the ways in which aging affects immune and gut function, and discusses clinical evidence for a role of dietary supplementation in countering age-related deficiencies in these processes. PMID- 11426288 TI - Nutritional issues and status of older persons of the Philippines: the IUNS, CRONOS and other studies. AB - The growing proportion of older persons in the Philippine population, with their potential contribution to development efforts as well as special needs for health and social services, presents an emerging demographic concern. The Philippines participated in the IUNS multicenter cross-cultural study of "Food Habits in Later Life" among urban elderly in three different care settings - elderly who lived with their families, those housed in government-operated homes for the aged, and older persons in private institutions. A second study employed the CRONOS protocol to differentiate nutritional and non-nutritional factors that affect the health and nutritional status of the Filipino elderly in urban and rural settings. Findings from the two studies revealed deficiencies in energy and protein intake among older persons. Those cared for by government social workers and care-givers had lowest protein-energy intakes and the highest proportion of underweight. Food habits of the urban elderly, particularly in terms of meal pattern, and use of table salt, sugar and fats were reported in the IUNS study. There was general perception of tiredness and limited social activity, network and support, but over-all self-rating of the elderly's health was good. The CRONOS study found significant association between later life status indicators, as well as between BMI and illness and physical activity scores. The paper included data from the 1993 National Nutrition Survey describing the nutritional situation of the elderly on a national scale: 30% of Filipino adults aged 60 y and over had a BMI <18.5 kg/m2 indicating chronic energy deficiency, 11% were obese with BMI >or=25 kg/m2, 41% were hypertensive and 45% anemic. Caring patterns were reflected in a 1984 four-country ASEAN study on the elderly which included the Philippines. While favorable, as the Filipino elderly generally lived in households where there was at least one adult female, the presence of other household members who likely competed for the time of the carer diminished the quality and adequacy of care for the persons. Further data is needed to update this information. The paper also discussed methodological and programmatic issues regarding improvement of the nutritional assessment of, and nutritional intervention programmes for the elderly. PMID- 11426289 TI - Nutritional factors on longevity and quality of life in Japan. AB - The relationship of nutrient intakes to life expectancies in Japan since the Second World War has demonstrated that sufficient intakes of animal protein and fat are crucial for attaining longevity. In the community dwelling elderly, the higher the serum albumin was, the longer the further life expectancy in the elderly. Serum total cholesterol showed a U-shape relationship to further life expectancies in the elderly. Low serum cholesterol was deleterious for higher levels of functional capacity. Low serum cholesterol and low serum alpha tochopherol accelerated depressive status in the community dwelling elderly. For reducing malnutrition and for improving nutritional status in undernourished elders, we carried out intervention trials. Instructions on how to prepare ideal daily meals were useful in functionally independent elders because a certain group of Japanese elderly has been stigmatized for eating meat. In frail elders provided with daily meals in a nursing home, a digestive enzyme drug was helpful for enhancing serum albumin levels. PMID- 11426290 TI - Chronic disease and dietary management in the elderly: alive and kicking! AB - The paper reviews progress in chronic disease prevention and dietary management strategies in the elderly to help them remain independent with a high quality of life for as long as possible. Progress in public health over the past century is briefly reviewed. The similarities and differences in preventive measures for elders and younger adults are summarized. The goals for the prevention and management of chronic disease in the elderly are similar, but the strategies may differ, and quality of life is especially important. Especially effective preventive strategies for the elderly are discussed. Selected strategies to manage conditions already present by delaying or controlling diet-related disease progression while maintaining an acceptable quality of life are discussed. Finally, selected new research on managing chronic disease in elders is reviewed. Taken together, these strategies promise to keep our elders "alive and kicking" for as long as possible. PMID- 11426291 TI - An assessment lexicon: assessment of dietary trends, physical activity patterns and nutritional status in the elderly. AB - This article discusses the use of nutritional assessment to improve the health and nutritional status of older persons. A three pronged approach is suggested. First, the emphasis is on screening for poor nutritional status. This involves a search for both biological and social factors that influence diet, physical activity and other determinants of nutritional status. The Determine Checklist is helpful in doing this. Second, after those elders potentially at risk are identified, the goal is to address risks to nutritional status more definitively. The nutritional status lexicon (ABCDEF's) must be assessed in this regard. Finally, it is critical to prevent or treat problems which have been discovered. Without follow-up,b screening and assessment are useless. PMID- 11426292 TI - Nutritional risk factors for institutional placement in Alzheimer's disease after one year follow-up. AB - Amongst factors associated with the institutional placement of elderly people with dementia, there has been little study of those related to malnutrition. We followed a cohort of 318 individuals with Alzheimer's disease (AD). Patients, who were all living at home at the start of the study were recruited from the outpatient service of a hospital unit specialising in AD. After one year, 20% of the patients had moved into institutional care. Multivariate analysis showed that a Mini nutritional Assessment score (MNA) of less than 25.5 (median score of the sample) and overeating behavioural problems (p=0.006) were risk factors for institutional placement. Nutritional problems are reversible and patients with a low MNA score could benefit from a thorough geriatric assessment, in order to slow or prevent institutional placement. PMID- 11426293 TI - Examining the effect of intervention to nutritional problems of the elderly living in an inner city area: a pilot project. AB - OBJECTIVE: To examine the effect of interventions provided to home-delivered meal service recipients living in an inner city neighborhood for the purpose of improving their nutritional conditions. DESIGN: Intervention study. SETTING AND PARTICIPANTS: Both the Study Group (42 seniors) and the Control Group (39 seniors), all of whom are over 60 years old, received home delivered meal services. In addition, only the Study Group received the following services through the trained neighborhood residents: 1) health education, 2) referrals to other community resources for unmet health needs, and 3) follow-up services to assure that the health problems were addressed appropriately. The nutritional condition, depression and cognitive status of the Study Group were assessed by medical staff before and after the six-month intervention. The same assessments were conducted to the Control Group with the same interval. Data was analyzed using ANOVA (repeated). MEASUREMENTS: MNA, GDS, and MMSE. RESULTS: Compared to the Control Group, the nutritional condition (p=0.052) and depression (p=0.02) of the Study Group were improved, while no significant difference was observed in their cognitive status. CONCLUSION: A combination of interventions by medical professionals and by trained neighborhood residents seemed to be an effective strategy to approach nutritional problems of seniors living in inner city areas. PMID- 11426294 TI - High prevalence and impact of subnormal serum vitamin B12 levels in Israeli elders admitted to a geriatric hospital. AB - BACKGROUND: The elderly are known to have higher rates of low and subnormal vitamin B12 levels than younger persons. Vitamin B12 deficiency has been extensively studied in the elderly, but primarily in outpatient settings. There is a paucity of data regarding the prevalence of low and low-normal B12 in frail, hospitalized, elderly patients, and its implications. Additionally, there is little information regarding vitamin B12 status in Israeli elders. OBJECTIVES: The objectives of the study were to estimate the prevalence of low and borderline vitamin B12 levels among frail, hospitalized, elderly patients, and their clinical implications. METHODS: We conducted a chart review, using a retrospective cohort design. The participants were 895 patients admitted to Harzfeld Medical Center in Gedera, Israel. Records were abstracted for vitamin B12 and Folic Acid levels, gastric disease, and outcomes including death, cognitive impairment and neurologic disease. RESULTS: Six hundred and forty patients were eligible for the study. In 15% of the patients, vitamin B12 level was in the low range (<150pmol/L) and in 25% in the low-normal range (150 250pmol/L). Gastric disease and antacid use were not associated with the vitamin B12 status. Mortality was higher in the high vitamin B12 group (p=0.02), perhaps reflecting a selection toward higher acuity in this group. Cerebrovascular disease was more common in patients with lower vitamin B12 levels (p=0.046). CONCLUSIONS: Forty percent of hospitalized elderly patients have low or borderline serum levels of vitamin B12, which may contribute to cerebrovascular disease and cognitive decline. PMID- 11426295 TI - Physiology of EAG K+ channels. PMID- 11426296 TI - Characterization of Na+-coupled glutamate/aspartate transport by a rat brain astrocyte line expressing GLAST and EAAC1. AB - D-aspartate (D-Asp) uptake by suspensions of cerebral rat brain astrocytes (RBA) maintained in long-term culture was studied as a means of characterizing function and regulation of Glutamate/Aspartate (Glu/Asp) transporter isoforms in the cells. A-asp influx is Na+-dependent with Km = 5 microm and Vmax = 0.7 nmoles x min(-1) x mg protein-1. Influx is sigmoidal as f[Na+] with Na+Km approximately 12 microm and Hill coefficient of 1.9. The cells establish steady-state D-Asp gradients >3,000-fold. Phorbol ester (PMA) enhances uptake, and gradients near 6,000-fold are achieved due to a 2-fold increase in Vmax, with no change in Km. At initial [D-Asp] = 10 microm, RBA take up more than 90% of total D-Asp, and extracellular levels are reduced to levels below 1 microm. Ionophores that dissipate the Delta(mu)Na+ inhibit gradient formation. Genistein (GEN, 100 microm), a PTK inhibitor, causes a 40% decrease in d-Asp. Inactive analogs of PMA (4alpha-PMA) and GEN (daidzein) have no detectable effect, although the stimulatory PMA response still occurs when GEN is present. Further specificity of action is indicated by the fact that PMA has no effect on Na+-coupled ALA uptake, but GEN is stimulatory. d-Asp uptake is strongly inhibited by serine-O-sulfate (S O-S), threohydroxy-aspartate (THA), L-Asp, and L-Glu, but not by D-Glu, kainic acid (KA), or dihydrokainate (DHK), an inhibition pattern characteristic of GLAST and EAAC1 transporter isoforms. mRNA for both isoforms was detected by RT-PCR, and Western blotting with appropriate antibodies shows that both proteins are expressed in these cells. PMID- 11426297 TI - Protein kinase A activation phosphorylates the rat ClC-2 Cl- channel but does not change activity. AB - Phosphorylation-dependent events have been shown to modulate the activity of several members of the mammalian CLC Cl- channel gene family, including the inward rectifier ClC-2. In the present study we investigated the regulation of rat ClC-2 expressed in the TSA-201 cell line (a transformed HEK293 cell line that stably expresses the SV40 T-antigen) by protein kinases. Protein kinase A activation phosphorylated ClC-2 in vivo, whereas stimulation of protein kinase C with phorbol 12-myristate 13-acetate did not. In vitro labeling studies confirmed that protein kinase A could directly phosphorylate ClC-2, and that protein kinase C and Ca2+/calmodulin-dependent protein kinase II did not. Nevertheless, protein kinase A-dependent phosphorylation of CLC-2 failed to regulate either the magnitude or the kinetics of the hyperpolarization-activated Cl- currents. Considered together, we demonstrate that protein kinase A activation results in the phosphorylation of rat ClC-2 in vivo, but this event is independent of Cl- channel activity. PMID- 11426298 TI - Regulation and properties of KCNQ1 (K(V)LQT1) and impact of the cystic fibrosis transmembrane conductance regulator. AB - The K+ channel KCNQ1 (K(V)LQT1) is a voltage-gated K+ channel, coexpressed with regulatory subunits such as KCNE1 (IsK, mink) or KCNE3, depending on the tissue examined. Here, we investigate regulation and properties of human and rat KCNQ1 and the impact of regulators such as KCNE1 and KCNE3. Because the cystic fibrosis transmembrane conductance regulator (CFTR) has also been suggested to regulate KCNQ1 channels we studied the effects of CFTR on KCNQ1 in Xenopus oocytes. Expression of both human and rat KCNQ1 induced time dependent K+ currents that were sensitive to Ba2+ and 293B. Coexpression with KCNE1 delayed voltage activation, while coexpression with KCNE3 accelerated current activation. KCNQ1 currents were activated by an increase in intracellular cAMP, independent of coexpression with KCNE1 or KCNE3. cAMP dependent activation was abolished in N terminal truncated hKCNQ1 but was still detectable after deletion of a single PKA phosphorylation motif. In the presence but not in the absence of KCNE1 or KCNE3, K+ currents were activated by the Ca2+ ionophore ionomycin. Coexpression of CFTR with either human or rat KCNQ1 had no impact on regulation of KCNQ1 K+ currents by cAMP but slightly shifted the concentration response curve for 293B. Thus, KCNQ1 expressed in Xenopus oocytes is regulated by cAMP and Ca2+ but is not affected by CFTR. PMID- 11426299 TI - Role of the S2 and S3 segment in determining the activation kinetics in Kv2.1 channels. AB - We constructed chimeras between the rapidly activating Kv1.2 channel and the slowly activating Kv2.1 channel in order to study to what extent sequence differences within the S1-S4 region contribute to the difference in activation kinetics. The channels were expressed in Xenopus oocytes and the currents were measured with a two-microelectrode voltage-clamp technique. Substitution of the S1-S4 region of Kv2.1 subunits by the ones of Kv1.2 resulted in chimeric channels which activated more rapidly than Kv2.1. Furthermore, activation kinetics were nearly voltage-independent in contrast to the pronounced voltage-dependent activation kinetics of both parent channels. Systematic screening of the S1-S4 region by the replacement of smaller protein parts resolved that the main functional changes generated by the S1-S4 substitution were generated by the S2 and the S3 segment. However, the effects of these segments were different: The S3 substitution reduced the effective gating charge and accelerated both a voltage dependent and a voltage-independent component of the activation time course. In contrast, the S2 substitution accelerated predominantly the voltage-dependent component of the activation time course thereby leaving the effective gating charge unchanged. It is concluded that the S2 and the S3 segment determine the activation kinetics in a specific manner. PMID- 11426300 TI - The composition-dependent presence of free (micellar) alkylphospholipid in liposomal formulations of octadecyl-1,1-dimethyl-piperidino-4-yl-phosphate affects its cytotoxic activity in vitro. AB - This study was performed to investigate the effect of cholesterol content, surface charge and sterical stabilization on the physico-chemical properties of liposomes prepared from the cancerostatic alkylphospholipid, octadecyl-1,1 dimethyl-piperidino-4-yl-phosphate (D21266), and their relationship to in vitro cytotoxicity. Stable incorporation of OPP into liposomes was found to be highly dependent on the cholesterol content. 31P-NMR spectroscopy as well as analysis of the lipid composition of OPP-containing liposome formulations revealed an increase in the amount of non-liposome-associated, micellar OPP as the cholesterol content decreased. The fraction of non-liposome-associated OPP constituted about 10% of total OPP when cholesterol was present in equimolar amounts (45.5/45.5 mol %) and increased to approximately 30% at a twofold excess of OPP over cholesterol (58.8/29.4 mol %). In monolayer incorporation studies it was shown that the existence of an increasing micellar pool of lipids leads to increased lipid transfer into the target monolayer. Liposome formulations containing more OPP than cholesterol were also found to display greater cytotoxicity. However, all liposome formulations were less cytotxic than pure (micellar) OPP. Cytotoxicity was not affected by the incorporation of N-methoxy polyethyleneglycol2000-phosphoethanolamine, a lipid that is known to reduce liposome uptake into phagocytic cells. The results demonstrate that the increase in cell toxicity correlates with the increase in non-liposome-associated, micellar OPP, which can readily exchange into cellular membranes. PMID- 11426301 TI - Effect of avermectins on Ca2+-dependent Cl- currents in plasmalemma of Chara corallina cells. AB - A natural complex of avermectins, aversectin C, and a component of this complex, avermectin A1, were shown to change the conductivity of Ca2+-dependent Cl- channels of plasmalemma of Chara corallina cells by acting from the outer side of the cellular membrane. Low concentrations of aversectin C and avermectin A1 increased the Cl- current: K1/2 = 35 ng/ml for the whole complex and K1/2 = 21 pg/ml for A1. Relatively high concentrations of the compounds suppressed the Cl- current: K1/2 = 2.2 microg/ml for aversectin C and K1/2 = 4.2 ng/ml for A1. The Hill coefficient for the interaction of avermectin A1 with the corresponding targets was identical for stimulation and suppression of the Cl- current: 2.8 and 2.5, respectively. Bicuculline, a nonspecific inhibitor of the GABAa receptors, did not influence stimulation of Cl- currents caused by low concentrations of avermectins, but at the same time blocked suppression of the Cl- currents by high concentrations of avermectins. Avermectins A2, B1, B2, abamectin and 22,23 dihydroavermectin B1 (ivermectin) did not affect the Cl- currents of Chara corallina cells. PMID- 11426302 TI - H+-ATPase-mediated cytoplasmic pH-responses associated with elevation of cytoplasmic calcium in cultured rabbit nonpigmented ciliary epithelium. AB - Studies were conducted to test whether an increase of cytoplasmic calcium concentration influences H+-ATPase activity in cultured rabbit nonpigmented ciliary epithelium (NPE). Cytoplasmic calcium concentration or cytoplasmic pH was measured by a fluorescence ratio technique in cells loaded with either Fura-2 or BCECF. Cytoplasmic calcium was increased in three ways; by exposure to BAY K 8644 (1 microm), by exposure to a mixture of epinephrine (1 microm) + acetylcholine (10 microm) or by depolarization with potassium-rich solution. In each case cytoplasmic pH increased significantly. In all three cases 100 nm bafilomycin A1, a specific H+-ATPase inhibitor, significantly inhibited the pH increase. These results suggest an increase of cytoplasmic calcium might initiate events that lead to activation of proton export from the cytoplasm by a mechanism involving H+-ATPase. This notion is supported by the observation that the pH increase was suppressed when either verapamil or nifedipine was used to prevent the cytoplasmic calcium increase in cells exposed to potassium-rich solution. Protein kinase C activation might also be involved in the mechanism of H+-ATPase stimulation since staurosporine suppressed the pH response to potassium-rich solution. A transient rise of cytoplasmic calcium concentration was observed when cytoplasmic acidification was induced by exposure to high pCO2. This suggests a rise of cytoplasmic calcium might represent part of a physiological mechanism to stimulate H+-ATPase-mediated protein export under acid conditions. PMID- 11426303 TI - Frontalis suspension technique with a temporal-fasciae-complex sheet for repairing blepharoptosis. AB - Congenital blepharoptosis presents as a drooping upper eyelid with an unpleasant appearance. Severe blepharoptosis usually has to be treated with varying modifications of frontalis suspension techniques. Each, however, has certain drawbacks. In this paper, a refined frontalis suspension technique with temporal fasciae-complex sheet is presented for repairing severe blepharoptosis. The technique suspends the eyelid from the frontalis muscle with a temporal-fasciae complex sheet through the submuscular tunnel between the eyelid and the frontalis. Each of the 16 patients (22 eyelids) achieved good upper eyelid elevation and proper palpebral crease formation. We found that the technique is simple, effective, and predictable with minimal risk of complications. It could be optimal for correcting severe blepharoptosis, especially for Oriental patients who demand palpebral crease formation. PMID- 11426304 TI - Full face rejuvenation in three dimensions: a "face-lifting" for the new millennium. AB - Traditional facial rejuvenation techniques address the face by lifting the soft tissues in one or two dimensions. The face is a tri-dimensional structure and aging occurs in three dimensions, therefore, facial rejuvenation should be done in three dimensions. Sagging of facial soft tissues occurs inferiorly and inferomedially. The ideal reorientation during rejuvenation is in the opposite direction: vertically and supero-laterally. Two other elements not routinely addressed by traditional rejuvenative operations are reduction of skeletal framework and atrophy of soft tissues, particularly subcutaneous fat layer. These are the third dimension of facial aging. By principle, any technique that unfolds, pulls, or lifts produces a flattening effect of the structure being treated. They may give a false impression of augmentation if these tissues are advanced over bony prominences. These stretched out tissues also have a tendency to recoil. For that reason, the author suggests use of structures or methods less susceptible to a stretch relaxation or recoil. A prerequisite to 3-D facial rejuvenation is to perform a 2-D-(bi-dimensional) lift. A third dimension is integrated into it. There are four methods to provide the third dimension: (1) augmentation of the skeletal framework; (2) augmentation of subcutaneous layer with fat injection; (3) imbrication of soft tissues; (4) mobilization and repositioning of fat pockets as pedicle flaps. These methods are not exclusive to each other. One, a few, or all methods could be integrated according to the patient's needs and aesthetic goals. Tridimensional changes in facial rejuvenation can be assessed by a standard photographic comparison, using tools for in-vivo measurements or 3-D digital imaging. 3-D facial rejuvenation is an advanced concept in our pursuit to provide superior results with the more aesthetic, natural, and harmonious youthful look to our patients. PMID- 11426305 TI - A successful long-term method of fat grafting: recontouring of a large subcutaneous postradiation thigh defect with autologous fat transplantation. AB - A method of harvesting fat with intact cells using a veterinary needle and a fine needle aspiration device is described. Viable fat cells are obtained in large amounts. A large subcutaneous postradiation thigh defect was reconstructed with harvested fat placed subcutaneously in various sessions. A good, but not perfect, reconstruction was obtained. This case indicates that viable fat cells can be transplanted and can survive even in the adverse environment of a postradiation defect. PMID- 11426306 TI - Carbon dioxide therapy in the treatment of localized adiposities: clinical study and histopathological correlations. AB - The authors report their experience using carbon dioxide (CO2) therapy for the treatment of 48 female patients presenting adipose accumulations, located on the thighs, knees, and/or abdomen; a Carbomed Programmable Automatic Carbon Dioxide Therapy apparatus was used. In light of the effects of CO2 on the microcirculation recently described in the literature, we expected this gas, which we administered subcutaneously, to positively affect the physiological oxidative lipolytic process. The aim of our study was to evaluate the effect of this therapy on localized adiposities. As such, we describe the method we used and report the results observed in the areas treated (in terms of reduction in maximum circumference) as well as side effects. Furthermore, we assessed the effect of subcutaneous administration of CO2 on the microcirculation by showing changes in the Laser Doppler signal and in the concentration of transcutaneous oxygen tension (tcPO2). Pre- and posttreatment biopsies of tissues were performed in seven patients in order to study the changes induced by the use of CO2 on both adipose and connective tissues. All data obtained were statistically analyzed; values of P < 0.05 were considered significant. PMID- 11426307 TI - Rhytidectomy assisted with ultrasound techniques: the ultra-lipo-lift technique. AB - Ultrasound-assisted liposuction has been practiced in Europe and South America for more than 10 years. This method was clinically introduced in 1989 for removal of fat by Dr. Zocchi, who developed the concept of applying ultrasound energy to adipose tissue [13,14]. Since 1992, we have been using ultrasound lipoplasty, first in body remodeling, then in face and neck surgery, after establishing working and safety rules in a study of 300 patients. In the beginning we were only looking to eliminate deposits of fat in oval faces with acne-scars, subdermal fibrosis, and thick skin, as these types of cases are not good candidates for rithidectomy. We found good skin retraction when we adjusted the superficial movements of the canula and had adequate machine power to avoid complications beneath the skin. Afterwards, we can break and release the adhesion zones of scarring and fibrosis, consequences of subdermic infiltrations or thread implants. Now we report our technique of rithidectomy assisted with ultrasound, which we have called ultra-lipo-lift (ULL). During the two last years we used this technique on 67 patients (48 females and 19 males) with good results and very few complications. PMID- 11426308 TI - Ultrasound assisted liposuction for the palliative treatment of Madelung's disease: a case report. AB - Benign Symmetric Lipomatosis (Madelung's disease) is a rare disease, characterized by massive fatty deposits in the neck, the shoulders, and the upper extremities. The deformity is associated with chronic alcohol use, malignant tumors of the upper airways, neuropathy, diabetes mellitus, hyperlipidemia, and other metabolic disorders. Although the deformity is prone to recurrence, surgical removal via lipectomy or liposuction provides the only way of palliation. This paper describes the treatment of a 51-year-old man with a history of alcoholism and liver cirrhosis. He reported masses in his cervical and facial regions that had gradually enlarged over a period of 6 years. He also developed respiratory symptoms due to the fatty compression of his upper airways. Our experience with ultrasound-assisted liposuction for the palliative treatment of this disease is reported. PMID- 11426309 TI - Wiping away debris between passes during laser surgery: is it really efficient or causing complications? AB - Although carbon dioxide laser skin resurfacing offers a controlled and predictive tissue ablation, it is not completely without complications, such as cobblestone like appearance or scar formation. It has been supposed that these were the results of thermal loading on overlapped areas. In order to obtain a smoother surface, an experimental study dealing with the effect of wiping procedure between the sessions was carried out. Laser beams were applied in two sessions to the backs of 20 rats. Between the sessions, the debris was wiped away in half and left unwiped in the others. Despite a statistically higher ablation rate in the wiped group, its surface irregularity was statistically significant when compared to unwiped group. It is supposed that the amount of debris left on the surface has the regulatory role in laser application. Because laser beams can be absorbed more by untreated skipped areas and less by overlapped sites, the second pass can ensure a smoother surface. Considering both benefits and disadvantages, it is concluded that wiping vaporized debris every two passes is the most reliable laser treatment modality. PMID- 11426310 TI - Gavello's procedure: an old earlobe reconstruction method, revisited and touched up. AB - Earlobe reconstruction following traumatic damage is a challenge for most of the plastic and reconstructive surgeons. Several methods developed for reconstruction of this deformity have shown to bear various advantages and disadvantages. Our modified Gavello's procedure, without using skin grafting in resurfacing the flap donor site, has shown to be satisfactory. This paper describes our modified operative procedure as well as a case resulting in a good looking earlobe and fine scars in flap donor site. PMID- 11426311 TI - An anatomical study of the medial canthus using a three-dimensional model. AB - Opinions on the direction and insertion of the muscle and tendon of the medial canthus not only differ depending on the reporter, but, to date, have lacked objectivity. The direction and insertion of the muscle and tendon of the medial canthus have, therefore, not been clear to surgeons operating on the medial canthus. In order to fully grasp the anatomy of this construct three dimensionality, we constructed a 3D model of successive sections of the medial canthus in a frontal direction using five cadavers, and then studied this model. The pretarsal part of the orbicularis oculi muscle is formed from a single muscle bundle of both the upper and lower eyelids, and runs into the medial palpebral tendon. This muscle bundle further branches off along the outside of the lacrimal sac, internally. It surrounds the back of the lacrimal sac without entering it. The preseptal part of the orbicularis oculi muscle consists of a single muscle bundle for both the upper and lower eyelids. The muscle fibers on the side of the skin run into the medial palpebral tendon. The muscle fibers posterior to this muscle bundle run into tendinous fibers, and, in all of the upper eyelids examined, they stop at the lacrimal fornix. In three out of the five lower eyelids examined the muscle fibers stop at the anterior surface of the lacrimal sac, while in the remaining cases they run into the medial palpebral tendon, as with the muscle fibers on the side of the skin. The medial palpebral tendon traverses the anterior surface of the lacrimal sac in an internal direction without branching off anteroposteriorly. PMID- 11426312 TI - Circumnipple-incision, longitudinal-breast dissection augmentation mammaplasty. AB - A modification of circumnipple breast augmentation is described, which confines the minimal scar to the base of the nipple. Furthermore, a longitudinal breast parenchyma dissection is performed for breast pocket dissection that limits the injury of the lactiferous ducts and nerves to a minimum, and makes dissection and exposure of the pocket easier, even in patients whose nipples are small. This technique is especially useful in small breasts. We have performed this procedure in 57 patients, and the few complications to date are discussed in this paper. PMID- 11426313 TI - Computed tomographic evaluation before cranial bone harvesting to avoid unexpected hazards during aesthetic procedures. AB - Nowadays, cranium is the preferred bone-graft donor site for facial aesthetic operations. Preoperative information about the quality of cranial bone, such as bone thickness or presence of the diplopic space, can be useful to minimize intracranial complications. This fact is neglected in reconstructive and aesthetic surgery. The aim of this study is to assess the reliability of Computed Tomography (CT) to determine cranial bone quality. Sixty-four cadaver parietal bones, the preferred site for bone-graft harvesting, were used in this study. In the first stage, posterior parietal bone thickness, which is accepted as the thickest part of cranium, was measured at specially determined points using a micrometer and the results were recorded. Bone thickness was then measured again in the same points with CT. The two methods were compared statistically. The measurements were not found to be statistically different. The similar values obtained with CT and micrometers suggest that CT can accurately and reliably determine cranial thickness. Preoperative CT can be a significant guide for the harvest of cranial bone grafts without any intracranial complications in aesthetic surgery. PMID- 11426314 TI - Effect of aesthetic rhinoplasty on respiratory functions. AB - Joseph's rhinoplasty operation, with minor modifications, was performed on 20 patients for aesthetic reasons. Various tests of the patient breathing though the nose were carried out both in the preoperative period and 10, 30, and 90 days after surgery. Respiratory-function test results improved in the majority of patients after rhinoplasty. FVC increased in 65% of the cases at day 10, in 85% at day 30, and in 75% at day 90. FEV1 increased in 65% of the cases at day 10, in 65% at day 30, and in 70% at day 90. FEV1/FVC increased in 60% of the cases at day 10, in 65% at day 30, and in 70% at day 90. The only statistically significant increase when compared with preoperative values was the increase in the values of FEV1/FVC at the 30th postoperative day. Decreases in respiratory function test values were insignificant in most of the patients. FEV1/FVC x 100 decreases of more than 10% were seen in three patients, indicating increased nasal resistance, at the 90th postoperative day. However, the lowest value was 50.4 and none of the patients reported subjective nasal obstruction. In conclusion it was observed in this study that properly executed cosmetic rhinoplasty did not have any significant negative effects on respiratory functions in most patients, and improved respiratory functions in most. PMID- 11426315 TI - The scientific contributions of British Plastic Surgeons to the Revue de Chirurgie Plastique and the Revue de Chirurgie Structive, Brussels (1931-1938). AB - In 1931, Maurice Coelst, M.D. from Brussels started the publication of the first international journal of plastic surgery ever published: the Revue de Chirurgie Plastique (1931-1934), which eventually became the Revue de Chirurgie Structive (1935-1938). In 1936, he established the first European Society of Structive Surgery, which held its first congress in Brussels. Further congresses were held in London in 1937 and in Milan in 1938. It is the collaboration and the participation of British plastic surgeons in this Society, this journal, and these meetings that I want to stress, because I am firmly convinced that these documents fill a gap in the history of Plastic Surgery in Great Britain, since- as far as I know--no detailed information concerning this period was ever published in Plastic Surgery literature. PMID- 11426316 TI - British plastic surgeons who contributed to the Revue de Chirurgie Plastique and the Revue de Chirurgie Structive (1931-1938): "the Big Four" in their Speciality. AB - The Revue de Chirurgie Plastique and the Revue de Chirurgie Structive, Brussels (1931-1938), edited by Maurice Coelst, M.D. from Brussels, were the first, full fledged medical publications devoted specifically to plastic, reconstructive, and aesthetic surgery. Publishing original articles by H.D. Gillies, P.T. Kilner, A.H. McIndoe, and R. Mowlem--the "Big Four" as they were known to both English and American plastic surgeons--the Revues drew attention to these four surgeons who were mainly responsible for developing the prestige of English plastic surgery in the early 1930s. PMID- 11426317 TI - Supernumerary nostrils: a case report and review. AB - Supernumerary or accessory nostrils are a very rare type of congenital nasal anomaly, with only a few cases reported in the literature. They can be associated with such malformations as facial clefts and they can be unilateral or bilateral, with most cases reported being unilateral. The accessory nostril may or may not communicate with the ipsilateral nasal cavity, probably depending on the degree of embryological progression of the anomaly. A case of simple supernumerary left nostril with no nasal cavity communication and with a normally developed nose is presented. The surgical treatment is described and the different speculative theories related to the embryogenesis of supernumerary nostrils are also reviewed. PMID- 11426319 TI - The correction of inverted nipples without scars. PMID- 11426320 TI - The human gene for mannan-binding lectin-associated serine protease-2 (MASP-2), the effector component of the lectin route of complement activation, is part of a tightly linked gene cluster on chromosome 1p36.2-3. AB - The proteases of the lectin pathway of complement activation, MASP-1 and MASP-2, are encoded by two separate genes. The MASP1 gene is located on chromosome 3q27, the MASP2 gene on chromosome 1p36.23-31. The genes for the classical complement activation pathway proteases, C1r and C1s, are linked on chromosome 12p13. We have shown that the MASP2 gene encodes two gene products, the 76 kDa MASP-2 serine protease and a plasma protein of 19 kDa, termed MAp19 or sMAP. Both gene products are components of the lectin pathway activation complex. We present the complete primary structure of the human MASP2 gene and the tight cluster that this locus forms with non-complement genes. A comparison of the MASP2 gene with the previously characterised C1s gene revealed identical positions of introns separating orthologous coding sequences, underlining the hypothesis that the C1s and MASP2 genes arose by exon shuffling from one ancestral gene. PMID- 11426321 TI - A population genetics study of single nucleotide polymorphisms in the interleukin 4 receptor alpha (IL4RA) gene. AB - Interleukin 4 (IL4) plays a critical role in T helper 2 (Th2) immune responses. Here we report a population genetics study of variation in the gene encoding the alpha-chain of the IL4 receptor (IL4RA) in three ethnic groups: African Americans, European Americans and East Asians. A 2941-bp region spanning exon 12 of IL4RA gene was sequenced in 12 individuals from each group. A total of 24 single nucleotide polymorphisms (SNPs) were identified in the combined sample. The genetic variation of the coding region of exon 12 is two to three times higher than in other reported genes. A significant departure from the expectation of evolutionary neutrality was observed, suggesting that natural selection may have influenced the evolution of this gene. We propose a model in which past selection by pathogens contributed to the increasing prevalence of atopic disorders in Western societies. PMID- 11426322 TI - Allele-specific quantification of tumor necrosis factor alpha (TNF) transcription and the role of promoter polymorphisms in rheumatoid arthritis patients and healthy individuals. AB - Interindividual variation in the expression of tumor necrosis factor alpha (TNF) suggests the existence of functionally distinct TNF alleles that could play a role in susceptibility to TNF associated diseases such as rheumatoid arthritis (RA). To determine whether differential expression of TNF alleles exists, the relative contribution of TNF alleles in total TNF RNA production in peripheral blood mononuclear cells (PBMC) of healthy individuals and synovial tissue of RA patients was analyzed. By using a Tai I restriction fragment length polymorphism (RFLP) located at position +489 in the first intron of the gene, the relative contribution of each allele in precursor transcript production in heterozygous individuals could be measured. By means of this method we studied whether differences exist between TNF alleles in TNF pre-mRNA production. The relative contribution of TNF alleles to the non-spliced RNA pool was measured in PBMC of healthy individuals which were stimulated with LPS, PMA and anti-CD3 and anti CD28 monoclonal antibodies for different time periods. Moreover, synovial biopsy material of RA patients was analyzed. The results of this study do not reveal a difference in the contribution of distinct TNF alleles in TNF pre-mRNA production upon in vitro and physiological stimulation conditions in healthy individuals and RA patients. Since some of the individuals whose PBMC were tested were also heterozygous for either -308, -1031, -863, -857 TNF promoter/enhancer single nucleotide polymorphisms (SNPs), the data argue against functional relevance of these TNF promoter/enhancer SNPs in the regulation of transcription. In conclusion, the data do not provide evidence for the existence of transcriptionally distinct TNF alleles to explain interindividual variation in TNF expression. PMID- 11426323 TI - CTLA-4 gene expression is influenced by promoter and exon 1 polymorphisms. AB - CTLA-4, expressed mainly on activated T cells, helps maintain, through its inhibitory function, immune-system homeostasis. Polymorphisms in the CTLA-4 gene (CTLA4) are known to be important in several autoimmune diseases, including multiple sclerosis (MS). Here, we have performed genotyping for CTLA4 polymorphisms, and investigated expression by peripheral blood mononuclear cells of CTLA-4 mRNA and protein, in patients with MS and myasthenia gravis and in healthy controls. Expression levels for mRNA and protein were similar in the patient and control groups; however, there was a clear relationship between genotype and CTLA-4 expression. Specifically, individuals carrying thymine at position -318 of the CTLA4 promoter (T(-318)) and homozygous for adenine at position 49 in exon 1 showed significantly increased expression both of cell surface CTLA-4 after cellular stimulation and of CTLA-4 mRNA in non-stimulated cells. The association was seen most clearly for unsorted CD3(+) cells and was absent in the CD8(+) subset. The T(-318) allele has been shown to be negatively associated with susceptibility to MS in an earlier study by our group. Thus, we propose that the susceptibility-influencing role of CTLA4 in MS may be related to genotypically conditioned promoter function, whereby high gene expression may decrease the risk of disease. PMID- 11426324 TI - Polymorphisms in the IkappaB-alpha promoter region and risk of diseases involving inflammation and fibrosis. AB - The transcription factor NFkappaB regulates inflammatory and other cellular responses. In non-stimulated cells, NFkappaB is linked to its inhibitor IkappaB, which plays a major role in controlling NFkappaB activity. Here, the gene promoter region of the major inducible IkappaB component (IkappaB-alpha) was studied to identify single nucleotide polymorphisms (SNPs), and to test if these are associated with risk of two diseases involving inflammation and fibrosis (trachoma and silicosis). Three SNPs were identified at positions -881, -826 and 297 relative to the transcription start site. The position -297 is close to two NFkappaB binding sites, kappaB2 and kappaB3, but the alleles were not associated with either disease. Alleles at positions -881 and -826 were in complete linkage disequilibrium with each other, and the rare haplotype was significantly less frequent among patients with trachoma compared to controls, although there was no difference in frequencies between silicosis patients and controls. PMID- 11426325 TI - Novel polymorphisms in human macrophage inflammatory protein-1 alpha (MIP-1alpha) gene. AB - Human macrophage inflammatory protein-1 alpha (MIP-1alpha) is a chemotactic cytokine, which binds to macrophages, T cells, and B cells affecting their activation. We found novel polymorphisms at four sites within MIP-1alpha gene in Japanese population: C to T in exon 2; A to G in intron 2; C to G and A to G in exon 3. They occurred on the same allele. Although MIP-1alpha effectively suppresses the replication of HIV-1 in vitro, we observed no statistically significant difference in the allele frequency of this polymorphism between HIV-1 infected and uninfected individuals in Japanese population. Since an increased transcription level of MIP-1alpha has been reported to be associated with inflammatory diseases such as atopic dermatitis, we also investigated the frequency of these polymorphisms among patients with atopic dermatitis, HIV-1 infected individuals (with a normal IgE level), and healthy donors. A small increase in ratio of homozygotes to other genotypes was observed in patients with atopic dermatitis (P = 0.04). PMID- 11426326 TI - Novel IFN-alpha receptor promoter polymorphisms. AB - We have identified two single nucleotide polymorphisms and one dinucleotide microsatellite within the promoter region of the interferon-alpha receptor gene (IFNAR, GenBank accession number X60459). Allele frequencies in a Caucasian population were established using PCR-RFLP for the two SNPs and semi-automated genotyping for the microsatellite. PMID- 11426329 TI - Vascular gene transfer driven by endoglin and ICAM-2 endothelial-specific promoters. AB - The involvement of the vascular endothelium in a large number of diseases supports the importance of vascular-specific gene delivery for their treatment. The hereditary hemorrhagic telangiectasia type 1 is an example of a vascular inherited disease (OMIM 187300). This is an autosomal dominant vascular disorder originated by mutations in the endoglin gene and associated with frequent epistaxis, telangiectases, gastrointestinal bleedings, and arteriovenous malformations in brain, lung and liver. Here, we address for the first time the possibility of using in vivo gene transfer to target endoglin expression to the vasculature. The promoter of the endothelial gene, ICAM-2, was used to generate transgenic animals which demonstrated endothelial expression of endoglin. Next, the promoters of the human endothelial genes, endoglin and ICAM-2, were inserted upstream of the human endoglin cDNA, and the resulting constructs were systemically or locally delivered, demonstrating endoglin expression in the vessel walls of liver, lung and skin. These gene transfer experiments represent an initial step in the treatment of the hereditary hemorrhagic telangiectasia type 1 by gene therapy, and suggest that endoglin and ICAM-2 promoters can be used to deliver other genes to the endothelium specifically. PMID- 11426330 TI - Intravenous injection of oligodeoxynucleotides to the NF-kappaB binding site inhibits hepatic metastasis of M5076 reticulosarcoma in mice. AB - We have developed synthetic double-stranded oligodeoxynucleotides (ODN) as 'decoy' cis elements that block the binding of nuclear factors to promoter regions of targeted genes, resulting in the inhibition of gene transactivation in vivo. In the present study, we employed decoy ODN targeting the transcription factor nuclear factor-kappaB (NF-kappaB) binding cis-elements to hepatic metastasis of murine reticulosarcoma M5076 in mice. Intravenous inoculation of M5076 into mice caused a marked increase in gene expression of interleukin-1beta, tumor necrosis factor-alpha and intercellular adhesion molecule-1 in the liver, whereas intravenous treatment with NF-kappaB decoy ODN reduced M5076-induced transactivation of these genes. Treatment with NF-kappaB decoy ODN, but not scrambled decoy ODN, significantly inhibited hepatic metastasis of M5076 in mice, and furthermore the combined treatment of NF-kappaB decoy ODN with an anti-cancer drug resulted in complete inhibition of hepatic metastasis in half of the mice, without affecting myelosuppression induced by the anti-cancer drug. Here, NF kappaB decoy ODN inhibited hepatic metastasis of M5076 in mice possibly through a decrease in transactivation of important NF-kappaB-driven genes and also potentiated the anti-metastatic effect of an anti-cancer drug, demonstrating the first successful in vivo therapy for cancer metastasis using NF-kappaB decoy ODN as a novel molecular decoy approach. PMID- 11426331 TI - Reduced growth in response to ganciclovir treatment of subcutaneous xenografts expressing HSV-tk in the vascular compartment. AB - Using a recombinant retrovirus with ecotropic envelope we have achieved high efficiency of transduction of endothelial cells in the vasculature of subcutaneous xenografts arising from the co-injection of tumour cells and irradiated virus producers. We have used this experimental system to assess the efficacy of the herpes simplex virus-thymidine kinase (HSV-tk)/ganciclovir (GCV) prodrug activation system in anti-vascular therapy. Treatment of KSY-1 xenografts with HSV-tk transduction in the vascular compartment with the S-phase-dependent drug GCV resulted in extensive haemorrhagic necrosis, indicative of vascular damage. Therapeutic potential in tumours with transduced endothelial cells comprising 5% or less of the total tumour mass was similar to that of tumours with HSV-tk expression in over 46% of tumour cells. GCV treatment of animals bearing MDA-MB-361 breast carcinoma, SW620 and CACO2 colon carcinomas with HSV-tk expression in the vascular compartment also resulted in reduced tumour growth. We conclude that HSV-tk/GCV prodrug activation is an effective strategy for eradicating tumour vasculature, and that direct targeting of proliferating endothelial cells in established vasculature results in reduced tumour growth. The therapeutic potential observed with the slow-growing CACO2 colon and MDA-MB 361 breast carcinomas supports the notion that anti-vascular therapy targeted at proliferating endothelium is likely to prove efficacious in human cancers that generally grow at a lower rate than experimental tumours. PMID- 11426332 TI - A novel MVMp-based vector system specifically designed to reduce the risk of replication-competent virus generation by homologous recombination. AB - Recent work highlights the potential usefulness of MVM-based vectors as selective vehicles for cancer gene therapy (Dupont et al, Gene Therapy, 2000; 7: 790-796). To implement this strategy, however, it is necessary to develop optimized methods for producing high-titer, helper-free parvovirus stocks. Recombinants of MVMp (rMVMp) are currently generated by transiently co-transfecting permissive cell lines with a plasmid carrying the vector genome and a helper plasmid expressing the capsid genes (replaced with a foreign gene in the vector genome). The resulting stocks, however, are always heavily contaminated with replication competent viruses (RCV), which precludes their use in vivo and particularly in gene therapy. In the present work we have developed a second-generation MVMp based vector system specifically designed to reduce the probability of RCV generation by homologous recombination. We have constructed a new MVMp-based vector and a new helper genome with minimal sequence overlap and have used the degeneracy of the genetic code to further decrease vector-helper homology. In this system, the left homologous region was almost completely eliminated and the right sequence overlap was reduced to 74 nt with only 61% homology. We were thus able to substantially reduce ( approximately 200 x), but not completely eliminate, generation of contaminating viruses in medium-scale rMVMp preparations. Since the remaining sequence homology between the new vector and helper genomes is weak, our results suggest that contaminating viruses in this system are generated by nonhomologous recombination. It is important to note, unlike the autonomously replicating helper viruses produced from the first generation vector/helper genomes, the contaminating viruses arising from the new packaging system cannot initiate secondary infection rounds (so they are not 'replication-competent viruses'). Our findings have important implications for the design of new MVMp-based vectors and for the construction of trans complementing packaging cell lines. PMID- 11426333 TI - Efficient infection of primitive hematopoietic stem cells by modified adenovirus. AB - Almost all studies of adenoviral vector-mediated gene transfer have made use of the adenovirus type 5 (Ad5). Unfortunately, Ad5 has been ineffective at infecting hematopoietic progenitor cells (HPC). Chimeric Ad5/F35 vectors that have been engineered to substitute the shorter-shafted fiber protein from Ad35 can efficiently infect committed hematopoietic cells and we now show highly effective gene transfer to primitive progenitor subsets. An Ad5GFP and Ad5/F35GFP vector was added to CD34(+) and CD34(-)lineage(-) (lin(-)) HPC. Only 5-20% of CD34(+) and CD34(-)lin(-) cells expressed GFP after Ad5 exposure. In contrast, with the Ad5/F35 vector, 30-70% of the CD34(+), 50-70% of the CD34(-)lin(-) and up to 60% of the CD38(-) HPC expressed GFP and there was little evident cellular toxicity. Because of these improved results, we also analyzed the ability of Ad5/F35 virus to infect the hoechst negative 'side population' (SP) of marrow cells, which appear to be among the very earliest multipotent HPC. Between 51% and 80% of marrow SP cells expressed GFP. The infected populations retained their ability to form colonies in two short-term culture systems, with no loss of viability. We also studied the transfer and expression of immunomodulatory genes, CD40L (cell surface expression) and interleukin-2 (secreted). Both were expressed at immunomodulatory levels for >5 days. The ability of Ad5/F35 to deliver transgenes to primitive HPC with high efficiency and low toxicity in the absence of growth factors provides an improved means of studying the consequences of transient gene expression in these cells. PMID- 11426334 TI - Efficient transduction of murine B lymphocytes and B lymphoma lines by modified adenoviral vectors: enhancement via targeting to FcR and heparan-containing proteins. AB - Murine lymphocytes are relatively refractory to efficient transfection or retroviral gene transduction. Adenovirus has been used as a vector to transduce a wide variety of cell types. Several advantages of adenoviruses are their ability to transduce non-cycling cells and to transduce the majority of cells in a population. Unfortunately, lymphocytes are not susceptible to infection with conventional adenovirus. Therefore, to express genes efficiently in murine B cells, we tested the ability of genetically modified adenovirus to transduce the beta-galactosidase gene. We found that adenovirus containing polylysine in the fiber knob was able to efficiently transduce lipopolysaccharide (LPS)-activated splenic B cells and the B lymphoma line M12.4.1; greater than 80% of the cells expressed beta-galactosidase activity. However, small resting B cells did not express activity unless treated with LPS after infection. This transduction was mediated by interaction with charged molecules since heparan-sulfate, and to a lesser degree chondroitan sulfate, inhibited the transduction. In addition, adenovirus containing a FLAG epitope in the fiber protein was used to target the FcR expressed on B cells using an anti-FLAG antibody. In the presence of anti FLAG, the modified adenovirus was able to efficiently transduce LPS-activated B cells and several B cell lymphoma lines. Interestingly, in the absence of anti FLAG, there was low level transduction in the LPS-blasts and in M12.4.1 that was not inhibited by soluble adenovirus fiber protein or agents that block RGD integrin interactions. These results demonstrate that modified adenovirus efficiently transduce B lymphocytes which will be critical for targeting genes to normal or malignant B cells. PMID- 11426335 TI - Inclusion of the herpes simplex thymidine kinase gene in a replicating adenovirus does not augment antitumor efficacy. AB - Replication-incompetent adenoviruses (Ad) carrying the herpes simplex thymidine kinase (HSVtk) gene have been used in a number of human cancer gene therapy trials, however transduction has generally been limited to a small minority of tumor cells. To solve this problem, replication-competent adenoviral vectors carrying transgenes such as HSVtk have been developed. However, contradictory evidence exists regarding the efficacy of these new vectors. Accordingly, we constructed and tested a replication-competent E3-deleted adenoviral vector containing the HSVtk suicide gene driven by the endogenous E3 promoter (Ad.wt.tk). This virus showed high level production of the HSVtk transgene and was more efficacious than a non-replicating virus in vitro, after injection into flank tumors, and against established intraperitoneal tumors. However, addition of ganciclovir (GCV) therapy to cells or tumor-bearing animals treated with the replicating vector containing the HSVtk suicide gene did not result in increased cell killing. Our results indicate that addition of HSVtk to a replicating Ad virus will not likely be useful in augmenting antitumor effects. PMID- 11426336 TI - Modulation of neuronal survival and axonal growth in vivo by tetracycline regulated neurotrophin expression. AB - Vector systems for the regulated and reversible expression of therapeutic genes are likely to improve the safety and efficacy of gene therapy for medical disease. In the present study, we investigated whether the expression of genes transferred into the central nervous system by ex vivo gene therapy can be regulated in vivo leading to controlled neuronal survival and axonal growth. Primary rat fibroblasts were transfected with a retrovirus containing a tetracycline responsive promoter for the expression of the neurotrophin nerve growth factor (NGF) or green fluorescent protein as a control (GFP). After lesions of basal forebrain cholinergic neurons, NGF-mediated neuronal rescue and axonal growth could be completely controlled over a 2-week period by the addition or removal of the tetracycline modulator doxycycline in the animals' drinking water. Further, continued expression of the reporter gene GFP could be reliably and repeatedly turned on and off in the injured CNS for at least 3 months post grafting, the longest time point investigated. These data constitute the first report of regulated neuronal rescue and axonal growth by controlled neurotrophin gene delivery and long-term, regulated expression using ex vivo CNS gene therapy. PMID- 11426337 TI - Expression of DeltaF508 CFTR in normal mouse lung after site-specific modification of CFTR sequences by SFHR. AB - The development of gene targeting strategies for specific modification of genomic DNA in human somatic cells has provided a potential gene therapy for the treatment of inherited diseases. One approach, small fragment homologous replacement (SFHR), directly targets and modifies specific genomic sequences with small fragments of exogenous DNA (400-800 bp) that are homologous to genomic sequences except for the desired modification. This approach has been effective for the in vitro modification of exon 10 in the cystic fibrosis transmembrane conductance regulator (CFTR) gene in human airway epithelial cells. As another step in the development of SFHR for gene therapy, studies were carried out to target and modify specific genomic sequences in exon 10 of the mouse CFTR (mCFTR) in vivo. Small DNA fragments (783 bp), homologous to mCFTR except for a 3-bp deletion (DeltaF508) and a silent mutation which introduces a unique restriction site (KpnI), were instilled into the lungs of normal mice using four different DNA vehicles (AVE, LipofectAMINE, DDAB, SuperFect). Successful modification was determined by PCR amplification of DNA or mRNA-derived cDNA followed by KpnI digestion. The results of these studies showed that SFHR can be used as a gene therapy to introduce specific modifications into the cells of clinically affected organs and that the cells will express the new sequence. PMID- 11426338 TI - Standard heparin, low molecular weight heparin, low molecular weight heparinoid, and recombinant hirudin differ in their ability to inhibit transduction by recombinant adeno-associated virus type 2 vectors. AB - Recombinant adeno-associated virus type 2 (rAAV) is a promising vector for in vivo gene therapy. Transduction by rAAV requires binding to heparan sulfate proteoglycan on the cell surface, and heparin can block this binding. Because heparin is administered to most patients undergoing cardiovascular gene transfer in order to prevent thrombotic events, it is important to identify anticoagulants which do not interfere with rAAV transduction. Therefore, we examined the influence of different anticoagulants on rAAV transduction in vitro. rAAV transduction was inhibited by 40.5 +/- 7.9% at heparin concentrations of 0.1 U/ml, and by 81.7 +/- 3.6% at 1.0 U/ml. The low molecular weight (LMW) heparin tinzaparin inhibited rAAV transduction by 20.2 +/- 3.8% at 0.1 U/ml and 37.1 +/- 1.8% at 1.0 U/ml. The inhibitory effect was significantly weaker compared with heparin at 1.0 U/ml, (P < 0.01). The LMW heparinoid danaparoid inhibited rAAV transduction by 8.8 +/- 3.5% at 0.1 U/ml (P < 0.01 compared with heparin). In contrast, recombinant hirudin did not interfere at all with rAAV transduction. In summary, the results demonstrate that inhibition of rAAV transduction by heparin occurs rapidly and at therapeutically used concentrations. LMW heparinoids and above all recombinant hirudin might be alternatives for heparin when vascular gene transfer with rAAV requires transient anticoagulation. PMID- 11426340 TI - Effects of diabetes on neurotransmission in rat vaginal smooth muscle. AB - The aim of this work was to characterize the effect of experimental diabetes on neurotransmission in rat vagina. Female Sprague-Dawley rats were divided into two groups: non-diabetic controls (NDM, n=38) and diabetics (DM, n=38). DM was produced by intraperitoneal injection of streptozotocin. Eight weeks later the animals were killed, the distal part of the vagina was removed, and smooth muscle strips were prepared for functional organ bath experiments and for measurement of nitric oxide synthase (NOS) activity. In DM preparations, the EC(50) value for noradrenaline (NA) was significantly increased (P<0.05) and the maximal contractile response decreased (P=0.001). In preparations precontracted with NA, the NO donor SNAP and calcitonin gene-related peptide (CGRP) caused concentration dependent relaxations, which were significantly decreased (P<0.001) in the DM group. Electrical stimulation of nerves (EFS) caused frequency-dependent contractions, which were significantly lower in DM than in NDM strips (P<0.001). SNAP and CGRP concentration-dependently inhibited EFS evoked contractions in both NDM and DM preparations. The inhibition was significantly lower (P<0.05) in the DM group. In NDM preparations precontracted with NA, EFS evoked frequency dependent relaxations; such relaxations were inhibited or reduced in DM. Treatment with the NOS inhibitor, L-NOARG 0.1 mM, abolished relaxations in all preparations or produced contraction in DM preparations. Calcium-dependent NOS activity was not significantly different in the DM and NDM groups. However, the DM animals showed a small but significant increase in calcium-independent NOS activity (P<0.05). Diabetes interferes with adrenergic-, cholinergic- and NANC neurotransmitter mechanisms in the smooth muscle of the rat vagina. The changes in the nitrergic neurotransmission are not due to reduction in NOS-activity, but seem to be due to interference with later steps in the L-arginine/NO/guanylate cyclase/cGMP system. PMID- 11426341 TI - RhoA/Rho-kinase: a novel player in the regulation of penile erection. AB - Current research has centered around the role of nitric oxide in the stimulation of cavernosal vasodilation and erection. However, recent evidence from our lab details the importance of endogenous vasoconstrictor mechanisms in maintaining a flaccid penile state, and further demonstrates that the inhibition of endogenous vasoconstriction is sufficient to stimulate erection in a rat model. In this article, we suggest inhibition of endogenous vasoconstriction as a potential therapeutic avenue in the treatment of erectile dysfunction. We also speculate on potential physiologic mechanisms by which endogenous vasoconstriction is inhibited in order for arousal-initiated vasorelaxation, and erection, to occur. PMID- 11426342 TI - A new potential of blood oxygenation level dependent (BOLD) functional MRI for evaluating cerebral centers of penile erection. AB - It is well known that penile erection is dependent on commands from the central nervous system. However, there has been little research on the central control of penile erection. The aim of this study was to evaluate, for the first time, the cerebral centers of penile erection using BOLD-functional MRI. Functional magnetic resonance imaging (fMRI) on a 1.5T MR scanner was performed in 12 sexually potent male volunteers (mean age: 23) and two hypogonadal impotent patients. In this study, blood oxygenation level dependent (BOLD) technique was utilized to create fMRI reflecting local brain activities. Real-time visual stimulation was performed with an alternatively combined erotic and non-erotic film to identify and quantify the activated brain regions associated with sexual response. Subjective sexual arousal and penile erection responses were assessed using 5-point scales ranging from 1 (no change) to 5 (maximal increase). In normal volunteers, the mean scores on subjective sexual arousal and penile erection by sexual stimulation with erotic film were 3.0 and 3.3 respectively, whereas there were no changes by non-erotic stimulation. During the visual stimulation the occipital cortex was activated by either an erotic or non-erotic film, the erotic film gave 150-200% stronger activation. However, more than seven of the 12 healthy subjects were significantly activated in the areas of inferior frontal lobe, cingulate gyrus, insula gyrus, corpus callosum, thalamus, caudate nucleus, globus pallidus, and inferior temporal lobe by erotic stimulation. In the hypogonadal patients, brain activation in response to the erotic film decreased compared to normal volunteers, however, it was restored by testosterone supplementation. These results are the first demonstration to show the functional neuroanatomy of the brain associated with sexual arousal by visual sexual stimulation using BOLD-based fMRI. Further studies are needed to verify that fMRI provides an important new tool in evaluating the cerebral center of the penile erection. PMID- 11426343 TI - Significant physiological roles of ancillary penile nerves on increase in intracavernous pressure in rats: experiments using electrical stimulation of the medial preoptic area. AB - The objectives of this work were to evaluate the contributions of the ancillary penile nerves to penile erection in male rats in vivo. We investigated the effects of unilateral and bilateral transection of the cavernous nerve (main penile nerve) on the increase in intracavernous pressure (ICP) following electrical stimulation of the medial preoptic area (MPOA) in male rats in vivo. After unilateral or bilateral transection of the cavernous nerve (main penile nerve), the ICP responses showed decreases of 28% and 55%, respectively compared to those ICP responses before transection. In other words, even after bilateral transection of the cavernous nerve, significant increases in the ICP response following central stimulation were observed. In contrast to these findings, the ICP response was completely eliminated following bilateral pelvic nerve transection. These data suggested that the ancillary penile nerves, which originate from the major pelvic ganglia, have a complementary role to the cavernous nerves in the autonomic motor innervation of the penis. PMID- 11426344 TI - Penile and perianal pudendal nerve somatosensory evoked potentials in the diagnosis of erectile dysfunction. AB - Neurophysiologic examinations in differential diagnosis of erectile dysfunction comprise electromyogramme of the pelvic floor, pudendal nerve terminal motor latency (PNTML) and evaluation of pudendal somatosensory evoked potentials (SSEP). We focused our interest on comparing diagnostic importance of penile and perianal pudendal nerve SSEP. We examined 20 patients suffering from erectile dysfunction and 20 patients without any manifestation of impotence. The stimulus was administered using penile ring electrodes at the base of the penis (cathode) and distally on the penis shaft (anode), as well as a perianal surface electrode applied at 3 o'clock in lithotomy position and 5 cm laterally on the gluteal skin. The potentials were recorded with intradermal needle electrodes at C(z)-2 cm (different) and F(z) (indifferent). 500 stimuli were averaged for a single tracing. The stimulus strength was set at an average of 3-4 times the stimulus threshold. Cortical latency of P 40 ranged from 39.0 to 45.6 ms (penile) and from 33.6 to 43.2 ms (perianal) in the control group, in the patient group latencies ranged from 38.8 to 51.6 (penile) and 34.0 to 44.8 ms (perianal). In two patients no potential was recordable after perianal stimulation, one patient showed a marked prolongation of the penile response with a normal perianal latency. Penile and perianal latencies of P 40 were significantly prolonged in the patient group compared to the control group (P<0.05). The combination of penile and perianal pudendal SSEP may provide valuable additional information in differential diagnosis of erectile dysfunction, especially allowing to identify different sites of neurogenic lesions. In contrast to perianal pudendal SSEP, penile stimulation may help to discover pathologic changes in the distal course of the pudendal nerve, especially the dorsal nerve of the penis. PMID- 11426345 TI - The effect of topically applied vasoactive agents and testosterone versus testosterone in the treatment of erectile dysfunction in aged men with low sexual interest. AB - The objective was to evaluate the efficacy and safety of topically applied cream containing testosterone, isosorbide dinitrate and co-dergocrine mesylate compared to testosterone cream in the treatment of erectile dysfunction in aged men with low sexual interest. A randomised double-blind crossover trial was performed over two months. The subjects were 42 men with erectile dysfunction and low normal or slightly depressed testosterone level randomly allocated to two equal groups. Polypharmacy cream containing testosterone 0.8%, isosorbide dinitrate 0.5% and co dergocrine mesylate 0.06% was applied for one month, and testosterone 0.8% cream for another month. The serum level of total testosterone was measured before and after each phase of treatment. Response to each therapy was assessed by a sexual questionnaire, measurement of tumescence and repeat penile duplex ultrasonography. Twenty-eight patients reported full erection and satisfactory intercourse with the polypharmacy cream. Thirteen men reported full erection and satisfactory intercourse with either cream. Polypharmacy cream increased penile arterial flow (P<0.001) and induced tumescence in 34 patients in lab. No patient in either phase of the study has tumescence or a significant increase in cavernous arterial peak systolic velocities after the application of testosterone cream. Serum level of total testosterone increased in all patients (P<0.05). Sexual desire was improved in 85% and 62% of patients during the treatment with polypharmacy cream and testosterone cream, respectively. No marked side effects were reported after either of them. Topical treatment with cream containing testosterone and vasoactive agents may represent a new effective treatment for erectile dysfunction associating with aging. PMID- 11426348 TI - Screening of ischemic heart disease with cavernous artery blood flow in erectile dysfunctional patients. AB - Erection is a hemodynamic event and accordingly, erectile dysfunction (ED) is closely related with ischemic heart disease. We should confirm that the cardiac condition of the ED patient is safe enough to perform sexual intercourse prior to beginning treatment for ED. Asymptomatic ischemic heart disease cannot be diagnosed only in an interview, but it's difficult to perform cardiac exercise tests on all patients complaining of ED. Therefore, screening methods to evaluate patients who should undergo exercise tests are needed. Sixty patients with erectile dysfunction participated in this study. Physical examinations, interviews, and color Doppler examinations were conducted. Chest X-rays and electrocardiograms of all patients in the resting position were obtained, as were electrocardiograms following exercise. Echocardiograms, treadmill test results, thallium exercise scintigrams, and coronary angiograms were obtained as required for diagnosis. Two patients were excluded because they had obvious arteriogenic ED due to perineal injury. Fifty-eight patients underwent Doppler evaluations of their cavernous arteries and heart exercise tests. Fourteen patients (24.1%) were diagnosed with ischemic heart disease. Although six of them had already been diagnosed with ischemic heart disease, eight were newly diagnosed by the exercise tests. Cardiovascular risk factors such as advanced age, hyperlipidemia, diabetes mellitus, hypertension, smoking, and obesity were not sufficient predictive factors. The mean peak systolic velocity of the patients without ischemic heart disease was 34.6 cm/s vs 22.0 cm/s in those with ischemic heart disease. Only 3.7% of patients whose peak systolic velocity in the cavernous artery was equal to or exceeded 35 cm/s had ischemic heart disease. On the other hand, 41.9% of patients with peak systolic velocity of less than 35 cm/s had ischemic heart disease. The sensitivity of peak systolic velocity against ischemic heart disease was 92.9%, and specificity was 59.1%. In ED patients, incidences of complications involving symptomatic or asymptomatic ischemic heart disease were found to be high. The peak systolic velocity in the cavernous artery is thought to be a useful predictive factor of ischemic heart disease in ED patients. When a patient reveals a peak systolic velocity of less than 35 cm/s, he should undergo heart exercise tests prior to treatment of ED. PMID- 11426349 TI - New insights into the role of endothelin-1 in radiation-associated impotence. AB - The objectives of this work were to: (1) Determine if prostate and penile tissue levels of endothelin-1 (ET-1) are increased in a rat following pelvic irradiation. (2) Determine if an ETa receptor antagonist (BQ-123) potentiates erectile function in these irradiated animals. Rats were divided into three study groups: control, 1000 cGy and 2000 cGy. The experimental groups received a single dose of radiation to the pelvic region. A time course was established to measure the effects of irradiation on prostate and penile tissue levels of endothelin-1 (ET-1)-like immunoreactivity. The effect of intracavernous injection of BQ-123 (25 microg/30 microl) was evaluated by measuring intracavernous pressure (ICP) following cavernous nerve electrical field stimulation. In the 2000 cGy group, a significant rise in ET-1-like immunoreactivity tissue levels was observed at 20 days. A significant decrease in ICP was recorded in the 1000 and 2000 cGy irradiated rats compared to the control group. Only the 2000 cGy group had a significant improvement in erectile function following BQ-123 administration. A significant improvement was observed 20 min post-administration, lasted 90 min, and was back to pre-administered levels at 120 min. The conclusion made was that radiation-induced impotence in irradiated rats is associated with an increased production of ET-1. Preliminary results are suggestive that ETa receptor antagonist may be of use to reverse such radiation-induced impotence in these irradiated animals. PMID- 11426350 TI - Spinal proerectile effect of apomorphine in the anesthetized rat. AB - Considering the presence of dopaminergic receptors in the lumbosacral spinal cord, we tested whether apomorphine could exert a proerectile effect by acting at the spinal level. Intracavernous (ICP) and blood pressures (BP) were measured in anesthetized rats. ICP rises were quantified (duration, percentage of ICPmaximum/meanBP (ICPmax/BPx100), area under ICP curve (AUC/BP) and sum of AUC/BP after intravenous (i.v.) and intrathecal (i.t.) injections of apomorphine alone or in presence of i.t. oxytocin (10 ng). Both 10 and 30 microg i.v. apomorphine dosings elicited erectile events evidenced by ICP rises. Upon the 30 microg i.v. injection, duration of ICP rises were increased from 25+/-10 to 69+/ 18 s (P<0.001), ICPmax/BPx100 from 21+/-3 to 50+/-14% (P=0.001), AUC/BP from 3+/ 1 to 14+/-6 s (P=0.002) and sum of AUC/BP from 5+/-7 to 34+/-35 s (P=0.021). Upon 30 microg i.t. injections of apomorphine at the lumbosacral level, the number of ICP rises was increased from 0.2+/-0.4 to 3.0+/-1.5, ICPmax/BPx100 from 16+/-9 to 43+/-12 and sum of AUC/BP from 1+/-3 to 31+/-15 s compared to vehicle injection (P<0.05 for all parameters). Injection of 30 microg i.v. or i.t. apomorphine non significantly enhanced the number and amplitude of the ICP rises induced by 10 ng i.t. oxytocin. However, the enhancement of the amplitude of the ICP rises elicited by i.t. oxytocin was more pronounced with i.t. apomorphine than with i.v. apomorphine. These results suggest the existence of a spinal site of action for apomorphine which may (1) participate to generation of erection and (2) exerts a facilitator effect on erection of supraspinal origin. PMID- 11426351 TI - Decreased circulating levels of estrogen alter vaginal and clitoral blood flow and structure in the rabbit. AB - Aging and menopause related decline in circulating levels of estrogen has been shown to adversely affect female sexual arousal function. Our aim was to study the effects of circulating levels of estrogen on the hemodynamic mechanism of vaginal and clitoral engorgement and on the structure of the vaginal and clitoral cavernosal tissue in the rabbit. New Zealand White female rabbits (3.5-4 kg) were randomly divided into three groups with five rabbits in each group: control; bilateral oophorectomy; bilateral oophorectomy undergoing subcutaneous injection of estrogen (40 microg/kg/day). After 6 weeks, the serum levels of 17 beta estradiol were measured and systemic blood pressure was monitored. Vaginal and clitoral cavernosal blood flows were measured with laser Doppler flowmeter before and after pelvic nerve stimulation. Cross sections of the clitoris and vagina were processed for histologic examination and histomorphometric image analysis. Serum level of 17 beta-estradiol (pg/ml; mean+/-s.d.) revealed a significant decrease in the oophorectomy group (25.4+/-5.1) compared with the control (38.5+/ 7.6) and estrogen replacement (115.9+/-57.3) groups (P<0.05). Nerve stimulation induced peak vaginal and clitoral intracavernosal blood flows in the oophorectomy group (28.9+/-16.3 and 6.1+/-1.4, respectively) were significantly less than those recorded in the control (48.9+/-6.5 and 11.0+/-2.4, respectively) or estrogen replacement (48.7+/-12.2 and 10.1+/-2.8, respectively) group (P<0.05). In histology, marked thinning of the vaginal epithelial layers, decreased vaginal submucosal microvasculature, and diffuse clitoral cavernosal fibrosis were evident in the oophorectomy group but not in the estrogen supplement and control groups. In histomorphometry, the percentage of clitoral cavernosal smooth muscle in the oophorectomy group (49.6+/-6.2) was significantly decreased compared with the control (56.8+/-2.6) and estrogen replacement (58+/-3.0) groups (P<0.05). Our studies show that decline in circulating levels of estrogen impairs the hemodynamic mechanism of vaginal and clitoral engorgement and leads to histopathologic changes in the vagina and clitoral cavernosal tissue. These observations suggest that decreased circulating levels of estrogen, a physiologic change in the menopausal state, may play a role in the development of female sexual arousal dysfunction. PMID- 11426352 TI - Sildenafil improves nocturnal penile erections in organic impotence. AB - We studied the effects of sildenafil on nocturnal penile erections. We prospectively evaluated 36 patients with organic or psychogenic impotence and 5 normal, potent men. All patients completed 3 sessions of consecutive nights using the RigiScan Plus device. The first two nights the patients were asked to take placebo before the session and to take 50 mg of sildenafil before the third session. In the organic impotence group the use of sildenafil induced a significant improvement in time of rigidity 60-100%, rigidity and tumescence activity unit values and rigidity and tumescence activity unit values per hour in the tip and base. In the psychogenic impotence group it caused significant improvement only in rigidity activity unit per hour in the tip. In the potent men, changes were statistically insignificant. Sildenafil improves nocturnal penile erectile activity in organic impotence. Our study shows that phosphodiesterase inhibitors can improve penile erections not induced by sexual stimulation. PMID- 11426353 TI - Clinical examination of the penis for fibrosis. PMID- 11426355 TI - Regional anesthesia for major cardiac and noncardiac surgery: more than just a strategy for effective analgesia? PMID- 11426356 TI - Pain relief after esophagectomy: Thoracic epidural analgesia is better than parenteral opioids. AB - OBJECTIVE: To compare postoperative pain relief and pulmonary function in patients after thoracoabdominal esophagectomy treated by continuing perioperative thoracic epidural anesthesia or changing to parenteral opioids. DESIGN: Prospective, randomized study. SETTING: University teaching hospital. PARTICIPANTS: Thirty-three patients undergoing thoracoabdominal esophagectomy. INTERVENTIONS: General anesthesia was combined with thoracic epidural anesthesia during surgery. The patients either continued with thoracic epidural analgesia (n = 18) or were switched to patient-controlled analgesia with intravenous morphine (n = 15) for 5 postoperative days. Pain scores were estimated twice daily, at rest and after mobilization. Peak expiratory flow, forced expiratory volume, and vital capacity were measured the day before surgery, postoperative day 2, and postoperative day 6. Adverse events and complications were recorded. MEASUREMENTS AND MAIN RESULTS: At rest, there were no differences in pain relief between the groups. Pain scores at mobilization showed a significantly lower value in the epidural group (p < 0.027). No intergroup differences were found regarding pulmonary function, which decreased on postoperative day 2, but was improved on postoperative day 6. CONCLUSION: Continuation of intraoperative thoracic epidural anesthesia for 5 postoperative days provides better pain relief at mobilization compared with a switch to patient-controlled analgesia with intravenous morphine. There was no intergroup difference in the impact on measures of pulmonary function. PMID- 11426357 TI - Comparison of continuous thoracic epidural and paravertebral blocks for postoperative analgesia after minimally invasive direct coronary artery bypass surgery. AB - OBJECTIVE: To compare continuous thoracic epidural analgesia (TEA) and paravertebral block (PVB) for postoperative analgesia in patients undergoing minimally invasive direct coronary artery bypass (MIDCAB) surgery for quality of analgesia, complications, compliance to chest physiotherapy, hemodynamics, and respiratory effects. DESIGN: Prospective, randomized study. SETTING: Specialty research hospital. PARTICIPANTS: Forty-one consenting patients undergoing MIDCAB surgery. INTERVENTIONS: Patients in the TEA group had an epidural catheter inserted at the T4-5 interspace, whereas patients in the PVB group had a catheter inserted in the paravertebral space on the left side at the T4-5 level. MEASUREMENTS AND MAIN RESULTS: Parameters evaluated included visual analog scale pain scores at rest and while coughing, supplemental analgesic requirement, complications, hemodynamics, and respiratory parameters. Measurements were made at 2-hour intervals for 12 hours beginning at 10 minutes after endotracheal extubation. There was no statistically significant difference in visual analog scale scores and requirement of supplemental analgesia between the 2 groups. Cardiac index at 4 hours and 6 hours was significantly higher in the TEA group. Patients in the PVB group had significantly lower respiratory rates at 8, 10, and 12 hours. All other parameters were comparable. In 1 patient, the epidural space could not be catheterized. One patient in the TEA group had transient hypotension, and 1 patient complained of backache at the site of the epidural catheter insertion. CONCLUSION: PVB is as effective as TEA for postoperative analgesia after MIDCAB surgery. PVB is technically easier than TEA and may be safer than TEA because no complications were seen in the PVB group. PMID- 11426358 TI - Transesophageal echocardiographic evaluation of native aortic valve area: utility of the double-envelope technique. AB - OBJECTIVE: To assess the accuracy of aortic valve area (AVA) calculations using the continuity equation with data obtained from the double envelope (DE) (simultaneously obtained left ventricular outflow tract [V1]) and aortic valve [V2] velocities) during intraoperative transesophageal echocardiography (TEE). DESIGN: Prospective study; measurements were performed on-line. SETTING: University hospital. PARTICIPANTS: Cardiac and noncardiac surgical patients (n = 75) with recent aortic valve assessment (<3 months) undergoing general anesthesia or endotracheal intubation. INTERVENTIONS: Intraoperative AVA was measured by the continuity equation using the DE technique (DE/TEE) and by planimetry (PL/TEE). Left ventricular outflow tract diameter was obtained from midesophageal views, whereas subvalvular (V1) and valvular (V2) velocities were obtained simultaneously using continuous-wave Doppler from transgastric views. V1 was also obtained using pulsed-wave Doppler. Measurements were compared with AVA obtained preoperatively by the Gorlin equation during cardiac catheterization (G/CATH) or by transthoracic echocardiography using the traditional continuity equation (C/TTE) (nonsimultaneously obtained V1 and V2). MEASUREMENTS AND MAIN RESULTS: A DE was obtained in 73 of 75 patients (97%). Four patients had atrial fibrillation at the time of the examination, whereas the rest were in sinus rhythm. PL/TEE was performed in 54 of 71 patients with sinus rhythm (76%). Agreement was good between DE/TEE and G/CATH (mean bias, 0.02 cm(2) [SD, 0.24 cm(2)]), and C/TTE (mean bias, -0.05 cm(2) [SD, 0.16 cm(2)]). Agreement was not as good between PL/TEE and G/CATH (mean bias, -0.07 cm(2) [SD, 0.28 cm(2)]) and C/TTE (mean bias, -0.13 cm(2) [SD, 0.30 cm(2)]). V1 obtained by pulsed-wave Doppler and with DE closely agreed (mean bias, 0.01 m/sec [SD, 0.05 m/sec]). CONCLUSION: TEE evaluation of native AVA using the DE technique is feasible and in good agreement with that obtained by C/TTE and G/CATH. Compared with DE/TEE, PL/TEE did not agree as well. Use of DE/TEE should simplify the continuity equation and may minimize errors resulting from beat-to-beat variability in stroke volume. PMID- 11426359 TI - The effects of the pericardium on length-dependent regulation of left ventricular function in coronary artery surgery patients. AB - OBJECTIVE: To analyze the effects of the pericardium on the length-dependent regulation of myocardial function in coronary artery surgery patients. DESIGN: Prospective. SETTING: University hospital. PARTICIPANTS: Patients scheduled for elective coronary artery surgery. INTERVENTIONS: In 10 patients, a combined micromanometer transducer conductance catheter was inserted into the left ventricle for measurement of left ventricular pressures and volumes. MEASUREMENTS AND MAIN RESULTS: Consecutive data were obtained during a progressive increase in left ventricular pressures and volumes obtained by leg elevation in closed chest closed pericardium and open chest-open pericardium conditions. Pericardiotomy did not alter baseline left ventricular hemodynamics. The effects of leg elevation were different, however. In closed chest-closed pericardium conditions, stroke volume and stroke work remained unchanged, whereas these parameters increased in open chest-open pericardium conditions. This increase was related to the increase in end-diastolic volume that was observed in open chest-open pericardium conditions and not in closed chest-closed pericardium conditions. CONCLUSIONS: In coronary artery surgery patients, pericardiotomy does not alter baseline left ventricular function. When cardiac load is increased by leg elevation, however, use of the Frank-Starling mechanism is enhanced in open chest-open pericardium conditions. PMID- 11426360 TI - Comparison of the hemodynamic effects of milrinone with dobutamine in patients after cardiac surgery. AB - OBJECTIVE: To compare the hemodynamic effects, efficacy, and safety of intravenous milrinone (M), 50 microg/kg during 10 minutes followed by 0.5 microg/kg/min, with intravenous dobutamine (D), 10 to 20 microg/kg/min, in patients with low cardiac output after cardiac surgery. DESIGN: Randomized, open label, multicenter study. SETTING: Cardiothoracic surgery departments, operating rooms, and intensive care units in 6 university hospitals. PARTICIPANTS: Patients (n = 120; 60 per group) after elective cardiac surgery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Analysis compared the hemodynamics at baseline and the percentage change from baseline during 4 hours of the drug infusion. The incidence of adverse events was recorded. Both groups had low mean (+/- SEM) cardiac indices (M, 1.6 ([0.03] L/min/m(2); D, 1.7 [0.03] L/min/m(2)) in association with adequate mean pulmonary capillary wedge pressures (M, 13.7 [1.3] mmHg; D, 12.7 [1.9] mmHg) at baseline. Group M had significantly higher systemic arterial pressures and systemic vascular resistances compared with group D; otherwise, the hemodynamics in both groups were comparable. During the study, hemodynamic responses included the following: group D had greater increases in cardiac index (at 1 hour, D = 55%, M = 36%; p < 0.01), heart rate (at 1 hour, D = 35%, M = 10%; p < 0.001), arterial pressures (mean arterial pressure at 1 hour, D = 31%, M = 7%; p < 0.001), and left ventricular stroke work index (at 1 hour, D = 75%, M = 45%; p < 0.05). Group M had greater decreases in mean pulmonary capillary wedge pressure (at 1 hour, D = -3%, M = -14%; p < 0.05). Comparisons of adverse events showed that dobutamine was associated with a higher incidence of hypertension (D = 40%, M = 13%; p < 0.02) and change of rhythm from sinus to atrial fibrillation (D = 18%, M = 5%; p < 0.04). Milrinone was associated with a higher incidence of sinus bradycardia (D = 2%, M = 13%; p < 0.03). CONCLUSIONS: Milrinone and dobutamine are appropriate and comparable for the pharmacologic treatment of the low- output syndrome after cardiopulmonary bypass. PMID- 11426361 TI - Influence of a new hydroxyethylstarch preparation (HES 130/0.4) on coagulation in cardiac surgical patients. AB - OBJECTIVE: To compare volume therapy with HES 130/0.4, a new hydroxyethylstarch (HES) solution with a gelatin-based fluid replacement strategy. DESIGN: Prospective, randomized, safety study. SETTING: Urban, university-affiliated hospital (single institution). PARTICIPANTS: Forty-two patients undergoing elective cardiac surgery. INTERVENTIONS: Patients were prospectively randomized into 2 groups: In group 1 (n = 21), gelatin was given perioperatively for volume support until the 1st postoperative day to keep the central venous pressure (CVP) between 10 and 14 mmHg; in group 2 (n = 21) HES 130/0.4 was administered using the same protocol as in group 1. MEASUREMENTS AND MAIN RESULTS: Standard coagulation variables and modified thromboelastography (TEG) were used. Using different activators for extrinsic and intrinsic activation and heparin inactivation by heparinase, the onset of coagulation (coagulation time), kinetics of clot formation (clot formation time), and maximum clot firmness were measured. Measurements were performed after induction of anesthesia (T0), at the end of surgery (T1), 4 hours after surgery (T2), and on the morning of the 1st postoperative day (T3). A total of 3310 +/- 810 mL of gelatin and 3070 +/- 570 mL of HES 130/0.4 were used in the 2 groups during the study period. The 2 groups did not differ with regard to postoperative bleeding or in use of packed red blood cells or fresh frozen plasma. Standard coagulation variables were similar between the 2 groups. All TEG variables were within the normal range at baseline. Coagulation time and clot formation time data were significantly elevated after surgery and in the intensive care unit, without showing specific differences between the 2 volume replacement groups. Intrinsic TEG and heparinase TEG clot formation times remained significantly higher until the end of the study period. No differences were seen between HES-treated and gelatin-treated patients. CONCLUSIONS: Volume replacement with the new HES preparation was as safe as gelatin-based volume replacement with regard to coagulation in cardiac surgical patients. HES 130/0.4 is an alternative plasma substitute to treat volume deficits. PMID- 11426362 TI - Blood loss in elective coronary artery surgery: a comparison of centrifugal versus roller pump heads during cardiopulmonary bypass. AB - OBJECTIVES: To compare the effects of centrifugal pumps versus roller pumps for cardiopulmonary bypass (CPB) in routine cardiac surgery on hematologic parameters in the context of modern practice. DESIGN: Prospective, randomized, partially blinded. SETTING: University teaching hospital. PARTICIPANTS: Elective coronary artery surgery patients (n = 113) INTERVENTION: Patients were randomized to be perfused with either a roller head (group R, n = 56) or a centrifugal head (group C, n = 57) pump. Patients received epsilon-aminocaproic acid before and during CPB. Core body temperatures were allowed to drift down to approximately 32 degrees C. MEASUREMENTS AND MAIN RESULTS: Postoperative chest tube blood loss, blood product requirements, hemoglobin, and platelet counts were assessed. There were no significant differences in preoperative or intraoperative parameters, including CPB time, complexity of procedure, and minimum core temperature. There were the expected reductions in hemoglobin and platelet levels post-CPB in both groups to a similar extent. Chest tubes remained in situ for similar durations, and the final volume of drainage was not significantly different (group C, 1300 +/- 92 mL; group R 1117 +/- 83 mL; p = 0.14). Allogeneic blood was given to 23% of patients in group C and 18% in group R (p = 0.63). Aspirin was associated with an increase in early chest tube drainage. CONCLUSIONS: In this surgical and perfusion environment, the authors were unable to show an advantage, from the hematologic point of view, in the routine use of a centrifugal pump head in elective coronary artery surgical patients. The use of antifibrinolytic agents and mild hypothermia may have effects on hemostasis that overshadow the influence of pump head design in this type of surgery. PMID- 11426363 TI - Comparison of the effects of a cell saver and low-dose aprotinin on blood loss and homologous blood usage in patients undergoing valve surgery. AB - OBJECTIVE: To compare 2 important techniques of blood conservation, use of a cell saver and low-dose aprotinin, in terms of blood loss and homologous blood usage in patients undergoing cardiac valve surgery. DESIGN: Prospective, randomized. SETTING: Tertiary care hospital. PARTICIPANTS: Sixty adult patients undergoing elective valve surgery. INTERVENTIONS: The patients were divided into 3 groups of 20 each. In group 1, aprotinin in the dose of 30,000 KIU/kg was added to the pump prime, with a further dose of 15,000 KIU/kg added at the end of each hour of cardiopulmonary bypass. In group 2, a cell-saver system was used to collect all blood at the operation site for processing in preparation for subsequent reinfusion. Group 3 patients acted as a control group and underwent routine management, which included collection of autologous blood during the pre cardiopulmonary bypass period. A hemoglobin of <8 g/dL was considered as an indication for bank blood transfusion in the postoperative period. MEASUREMENTS AND MAIN RESULTS: The chest tube drainage was significantly less in group 1 compared with groups 2 and 3, with total drainage (median [interquartile range]) amounting to 250 mL [105 to 325 mL] vs. 700 mL [525 to 910 mL] in group 2 and 800 mL [650 to 880 mL] in group 3 (p < 0.001). The patients in groups 1 and 2 required significantly less bank blood (median [interquartile range]) as compared with group 3 (350 mL [0 to 525 mL], 350 mL [0 to 350 mL], and 1050 mL [875 to 1050 mL]; p < 0.001), respectively. Cell saver provided 410 +/- 130 mL of hemoconcentrated blood in group 2. The average preoperative hemoglobin concentration was 11.3 g/dL, and it was around 9 g/dL on the 7th postoperative day. The hemoglobin concentration at various stages during hospitalization in all 3 groups was similar. CONCLUSIONS: Low-dose aprotinin and a cell saver are effective and comparable methods of blood conservation. Aprotinin helps by decreasing the postoperative drainage, and a cell saver helps by making the patient's own blood available for transfusion. PMID- 11426364 TI - Tranexamic acid in aortic valve replacement. AB - OBJECTIVE: To assess the relative efficacy of tranexamic acid compared with a control group to decrease bleeding and transfusion requirements in a uniform population undergoing aortic valve replacement. DESIGN: Prospective, randomized, double-blind study. SETTINGS: University hospital. PARTICIPANTS: Adult cardiac surgery patients (n = 300). INTERVENTIONS: Patients were randomized into 2 groups to receive either a total of 5 g of tranexamic acid or a saline solution. Bleeding in the postoperative period, transfusions of bank blood products, coagulation profile, intensive care unit stay, and hospital length of stay were recorded. MEASUREMENTS AND MAIN RESULTS: Postoperative bleeding in patients treated with tranexamic acid was significantly lower compared with the control group (p < 0.0001). Packed red blood cells and fresh frozen plasma usage were reduced in the tranexamic acid group compared with the control group (p = 0.0095 and p < 0.0001). Only 24.5% of tranexamic acid patients received blood products versus 45% of control patients (p < 0.01). There was no significant difference in hematologic and coagulation profiles after the operation between the groups. CONCLUSIONS: Tranexamic acid reduces postoperative blood loss and transfusion requirements in elective aortic valve replacement. PMID- 11426365 TI - Influence of a vital capacity maneuver on pulmonary gas exchange after cardiopulmonary bypass. AB - OBJECTIVE: To investigate the effect of a single, vital capacity breath (vital capacity maneuver [VCM]), administered at the end of cardiopulmonary bypass (CPB), on pulmonary gas exchange in patients undergoing coronary artery bypass graft surgery. DESIGN: Prospective, randomized, double-blind study. SETTING: University-affiliated hospital. PARTICIPANTS: Forty patients scheduled for elective coronary artery bypass graft surgery and early tracheal extubation. INTERVENTIONS: Patients were randomized to 1 of 2 groups. VCM patients received a VCM at the conclusion of CPB. Control patients received no VCM. MEASUREMENTS AND MAIN RESULTS: Intrapulmonary shunt (Q(S)/Q(T)), arterial oxygenation (PaO2), and alveolar-arterial oxygen gradients (P(A-a)O2) were measured after induction of anesthesia, CPB, intensive care unit (ICU) arrival, and extubation. The duration of postoperative intubation was recorded for each group. Q(S)/Q(T) increased significantly 30 minutes after CPB in the control group (15.7 +/- 1.8% to 27.4 +/ 2.6%; p = 0.01). In the VCM group, a small decrease in Q(S)/Q(T) occurred (16.1 +/- 2.0% to 14.9 +/- 2.0%). After ICU arrival and extubation, no significant difference in Q(S)/Q(T) existed between the 2 groups. With the exception of a higher P(A-a)O2 in the control group at induction of anesthesia, no differences in PaO2 or P(A-a)O2 were present between the 2 groups at any measurement interval. Patients who received a VCM were extubated earlier than the control group (6.5 +/- 2.1 hours v 9.4 +/- 4.2 hours; p = 0.01). CONCLUSION: The use of a VCM prevented an increase in Q(S)/Q(T) from occurring in the operating room. Although a VCM did not influence pulmonary gas exchange in the ICU, its application in the operating room appears to exert a beneficial effect on tracheal extubation times after cardiac surgery. PMID- 11426366 TI - Techniques and complications of one-lung ventilation in children with suppurative lung disease: experience in 15 cases. AB - OBJECTIVE: To evaluate lung isolation with Fogarty catheters and to analyze respiratory consequences of one-lung ventilation (OLV) in children with suppurative lung disease. DESIGN: Prospective. SETTING: University hospital. PARTICIPANTS: Fifteen children undergoing thoracotomy. INTERVENTIONS: Bronchial blockade with a 7F Fogarty catheter was attempted. In case of incomplete blockade or failure in directing the catheter into the desired mainstem bronchus, endobronchial intubation was done. Volume-controlled ventilation was performed with fraction of inspired oxygen (F(I)O2), 0.5; inspiratory-to-expiratory (I: E) ratio, 1:2; and 10 mL/kg tidal volume during two-lung ventilation (TLV). F(I)O2 was increased to 1.0 by the initiation of OLV. If peak airway pressure exceeded basal values during TLV by 35%, tidal volume was reduced to 8 mL/kg, inspiratory pause was zeroed, and I:E ratio was increased to 1:1. Hemodynamic and respiratory parameters were recorded during TLV and 30 minutes after initiation of OLV. Peripheral oxygen saturation and end-tidal carbon dioxide tension were recorded every 5 minutes. MEASUREMENTS AND MAIN RESULTS: Right lung isolation was successfully obtained by Fogarty catheters in 10 children undergoing right thoracotomy. Endobronchial intubation was performed in 2 children (40%) undergoing left thoracotomy. Three children (20%) developed episodes of severe hypercapnia and hypoxia requiring treatment during OLV. All of the parameters recorded at 30 minutes of OLV revealed statistically significant differences from TLV. OLV was transiently discontinued in 1 child. CONCLUSION: The use of Fogarty embolectomy catheters for lung isolation in children undergoing thoracotomy is recommended. Respiratory problems are not rare during OLV in children with suppurative lung disease and require immediate management. PMID- 11426367 TI - Repeated administration of protamine does not attenuate circulatory changes caused by protamine reversal of heparin in dogs. AB - OBJECTIVE: To determine whether repeated administration of protamine attenuates circulatory changes caused by protamine reversal of heparin and to evaluate the significance of nitric oxide generation. DESIGN: Prospective, randomized, controlled, animal study. SETTING: University research laboratory. PARTICIPANTS: Twenty-seven adult mongrel dogs. INTERVENTIONS: The animals were randomly assigned to 3 groups (n = 9 in each) according to the pretreatment. The control group was pretreated with normal saline, and the 2 other groups were given 2 different doses of protamine: protamine 0.1 (protamine, 0.1 mg/kg) and protamine 1.0 (protamine, 1.0 mg/kg). Under general anesthesia, all animals were anticoagulated with intravenous heparin, 200 IU/kg. Five minutes after heparin injection, preadministered saline (control) or protamine in saline was infused during 60 seconds. Five minutes after the pretreatment, protamine, 2.0 mg/kg in control, 1.9 mg/kg in protamine 0.1, or 1.0 mg/kg in protamine 1.0, was administered intravenously during 10 seconds. MEASUREMENTS AND MAIN RESULTS: Percent changes in mean arterial blood pressure among the 3 groups at each period were not significantly different except 60 minutes after protamine infusion. Mean pulmonary arterial pressure in the protamine 1.0 group at 5, 15, 20, and 60 minutes was higher than in the control group. Serum nitrate concentration was not significantly different among the 3 groups at baseline and 10 minutes after protamine injection. CONCLUSION: Repeated administration of protamine does not attenuate circulatory changes caused by protamine reversal of heparin in dogs. Nitric oxide generation does not appear to be responsible for the phenomenon. PMID- 11426368 TI - Off-pump coronary artery bypass surgery in a patient with a symptomatic intracranial mass. PMID- 11426369 TI - Severe dental pain during carotid endarterectomy under cervical plexus block. PMID- 11426370 TI - Hypogammaglobulinemia complicating chylothorax after cardiac surgery in two infants. PMID- 11426371 TI - Anesthetic management of a patient with hypertrophic obstructive cardiomyopathy after percutaneous transluminal septal myocardial ablation: early postinterventional period and anesthetic considerations. PMID- 11426372 TI - Myocardial dysfunction in septic shock: Part I. Clinical manifestation of cardiovascular dysfunction. PMID- 11426373 TI - Case 6--2001: Exsanguinating endotracheal hemorrhage during cardiopulmonary bypass. PMID- 11426374 TI - Pro: Only pediatric anesthesiologists should administer anesthetics to pediatric patients undergoing cardiac surgical procedures. PMID- 11426375 TI - Con: Anesthetics should be administered to pediatric patients undergoing cardiac surgical procedures not only by pediatric anesthesiologists. PMID- 11426376 TI - Pro: Pediatric anesthesiologists should be the primary echocardiographers for pediatric patients undergoing cardiac surgical procedures. PMID- 11426377 TI - Con: Pediatric anesthesiologists should not be the primary echocardiographers for pediatric patients undergoing cardiac surgical procedures. PMID- 11426378 TI - Echocardiographic diagnosis of left ventricular density: diagnostic pitfalls. PMID- 11426379 TI - An unusual finding in a patient with hypertrophic cardiomyopathy. PMID- 11426380 TI - "Baffled" by the conclusions. PMID- 11426382 TI - Hyperglycemia: an independent predictor of cardiovascular risk. PMID- 11426383 TI - Management of complex defects with pulmonary hypertension. PMID- 11426385 TI - Iatrogenic intrapericardial diaphragmatic hernia associated with cardiac tamponade. PMID- 11426386 TI - Surfactant-immobilized fibronectin enhances bioactivity and regulates sensory neurite outgrowth. AB - A PEO-containing surface coating was investigated as a means to control neurite outgrowth in the presence of serum. Various ratios of end-group-activated tri block copolymer Pluronic F108 were used to immobilize the extracellular matrix protein fibronectin (FN). Primary cultures of dorsal root ganglion neurons were cultured on F108-immobilized FN or, as a control, on FN adsorbed from solution directly to polystyrene. Although FN surface concentration could be controlled in a dose-dependent manner by either technique, dose-dependent control of neuronal behaviors was best achieved on F108-immobilized FN. This effect was similar regardless of the presence of serum in the culture medium. F108-immobilized FN supported twofold greater maximal neurite outgrowth than did directly adsorbed FN. Furthermore, at similar surface concentrations, F108-FN was significantly more active in promoting neurite outgrowth. Polypropylene filament bundles treated with F108-immobilized FN supported robust outgrowth from explants of dorsal root ganglia, demonstrating the utility of the surface coating on clinically relevant materials with more complex shapes. The ability to control neuronal behaviors in a serum-resistant manner, coupled with enhanced biologic activity, demonstrates the potential for surfactant-based immobilization as a method for generating biointeractive materials for tissue engineering. PMID- 11426387 TI - Fibronectin facilitates adhesion of K562 leukemic cells normally growing in suspension to cationic surfaces. AB - The role of protein adsorption in the forced adhesive growth of K562 leukemic cells onto a cationic surface composed of polylysine was investigated. Numerous studies have demonstrated that adhesion in anchorage-dependent cells is mediated in vitro by adsorption of serum proteins [particularly proteins of the extracellular matrix (ECM) such as fibronectin and vitronectin] present in the growth medium. Specifically, adhesion has been shown to occur when ECM proteins attach to the substratum and act as ligands for specific receptors located on the surface of cells. K562 cells are human erythroleukemic cells that normally grow in suspension. These cells are not involved in the same cell adhesion processes as anchorage-dependent cells and do not need to be attached to ECM proteins in order to survive and grow. Thus, with these systems, it is possible to better determine the role of protein adsorption in the adhesion of cells, growing in suspension such as blood cells, onto charged surfaces. The results presented show that adhesion of K562 cells onto the positively charged polylysine surface in the presence of serum is mediated through specific interactions between fibronectin receptors present on K562 cells and fibronectin adsorbed onto that cationic surface. Specifically, determination of cell adhesion under different experimental conditions indicates that nonspecific charge interactions do not take place directly between the cells and polylysine, but rather take place between polylysine and fibronectin, which adsorbs onto the cationic polymer. In addition, flow cytometric analyses reveal that only fibronectin receptors are present on these cells and, consequently, only fibronectin can be responsible for the actual adhesion of these cells onto the cationic surface. In view of the data presented, the possibility should be considered that ECM components adsorbed onto surfaces with specific charges and/or belonging to certain functional groups are involved in structural and functional modifications in cells. These cells grow in suspension and are normally not involved in adhesion phenomena, though these components should be considered. These considerations should be made especially when designing biomaterials that can modulate the response of cells growing in suspension, such as blood cells, and also in tissue engineering of blood substitutes. PMID- 11426388 TI - High mass clearance of autoantibodies from a murine model of lupus nephritis by immunoadsorption using star-configured polyethylene glycols. AB - The extracorporeal immunoadsorption of antibodies as part of the therapy for human autoimmune diseases has been limited by technology with inadequate and nonselective mass clearance or problems with bioincompatibility. To overcome these shortcomings, we designed a method utilizing star-configured polyethylene glycols (star-PEGs) having up to 63 free arms with immunoreactive (tresylate ester) end-groups for each arm immobilized to a polymer support substrate. The flexibility and length of the arms are thought to allow optimization of epitope presentation and to permit interaction with immunoligands on adjacent arms. To demonstrate efficacy we used an in vitro murine antibody model of human lupus nephritis, wherein we could study the kinetics and mass clearance of hybridoma derived antihistone antibodies from human plasma. Histones were covalently bound to the star-PEG end-groups and the kinetics of antibody adsorption were assessed using a surface plasmon resonance technique. The equilibrium constants of antihistone antibody binding to histone-star-PEGs that were linked to a support grid demonstrated high affinity with a KA of 3.56E + 07 and a KD of 2.81E - 08. The optimum reaction conditions were determined to accomplish the hydrophilization of polysulfone (PS; by an aqueous nitration method) and polymethylmethacrylate substrates (PMMA; by hydrazine), using sheet casts of both polymer substances. Hollow fiber devices of these polymers (commercial hemodialyzers) were modified so that histone-bound star-PEGs were linked to their intracapillary luminal surfaces, using a process which we have shown retains their immunoadsorption properties for antihistone antibodies. A closed loop recirculating model was constructed to measure mass clearance of antibodies from a reservoir. After optimizing conditions using extraction from saline solutions, the removal of antibody from human plasma by control and surface-modified devices was assessed over 4 h. There was no measurable antibody clearance by the control fibers over this time interval. The 2.1 m2 luminal surface area PMMA devices removed 5.0 +/- 1.1 mg, with a maximum of 7.0 mg. The 1.8 m2 PS device cleared 11.3 +/- 6.2 mg with a maximum of 17.5 mg. In summary, star-PEG immunoadsorption is a promising technique for the treatment of human autoimmune disease because it can achieve very high-mass clearance of autoantibodies using modified biocompatible hollow-fiber polymer devices. PMID- 11426389 TI - Investigations of thermal property gradients in the human dentine. AB - An investigation of the adaptation of dentine to temperature variation was conducted with the aid of digital moire interferometry and thermomechanical analysis. The moire interferometric patterns provided direct evidence of two major phases of thermally involved deformation in dentine. An initial phase of expansion was followed by contraction at higher temperatures. Significant gradients in thermal strain and the coefficient of thermal expansion were identified. Close agreement was found between the response of dentine to thermal changes as observed by moire interferometry and that detected by thermomechanical analysis. This study highlights the biological adaptation of dentine to thermal variations. PMID- 11426390 TI - Effects of hyaluronan on engineered articular cartilage extracellular matrix gene expression in 3-dimensional collagen scaffolds. AB - Hyaluronan (HA) is a component of cartilage matrix with known effects on chondrocytes. We tested the effects of adding HA to 3-dimensional (3-D) collagen. sponges on chondrocyte function in vitro. Bovine articular chondrocytes isolated by collagenase digestion were injected into either collagen or HA/collagen scaffolds comprising different amounts of HA (2, 5, 10, and 14% w/w). Expression of aggrecan and type II collagen genes was measured by gene-specific quantitative competitive reverse transcriptase-polymerase chain reactions, and the extracellular matrix was estimated by histomorphometrical analyses. After 7-day culture, the chondrocytes in 2% (w/w) HA sponges expressed fourfold more mRNA transcripts for type II collagen (p = 0.002) and twofold more mRNA transcripts for aggrecan (p = 0.022) than in control collagen sponges. Furthermore, there was 45% more extracellular matrix in 2% (w/w) HA sponges and 43% less matrix in the 10% (w/w) HA sponges compared with plain collagen sponges (p > 0.05). In sum, a small amount of HA in 3-D collagen scaffolds enhanced chondrogenesis, but a greater amount was inhibitory. This 3-D system represents a novel tool to identify mechanisms by which extracellular matrix molecules influence chondrocyte function. Further, these results show the potential for modifying scaffolds to improve production of engineered cartilage for in vivo applications. PMID- 11426391 TI - Evaluation of tissue-factor production by human endothelial cells incubated with three acrylic bone cements. AB - The effect of three methacrylate-based cements used for the fixation of joint prostheses on tissue-factor production by human umbilical vein endothelial cells was evaluated in vitro. The extracts in the culture medium of the cements were tested after 1-h and 7-day curing. The endothelial cells were incubated with the cement extracts for 4 h, and then the tissue factor was determined in cell lysates with both the recalcification time and enzyme immuno assay. The cements did not induce significant production of tissue factor and, therefore, did not activate the extrinsic pathway of coagulation within the limits of the mechanism considered. PMID- 11426392 TI - Novel patterned cell coculture utilizing thermally responsive grafted polymer surfaces. AB - Here we demonstrate a novel cell coculture method without any apparent limitation in cell-type combinations that exploits thermally responsive polymer-grafted patterns to alter cell-cell and cell-surface interactions. Thermally responsive acrylamide polymer is first covalently patterned onto culture surfaces by masked electron beam irradiation. One cell type is then cultured to confluency at 37 degrees C. Reducing cell culture temperature below 32 degrees C selectively swells temperature sensitive polymer-grafted domains, detaching adherent cells only from these grafted patterns. Another cell type is then seeded over the same surface at 37 degrees C. These subsequently seeded cells adhere only to the now exposed polymer-grafted domains. Initially seeded cells remaining adherent on nonpatterned surfaces and cells added in the second seeding are then cocultured at 37 degrees C in well-ordered patterns. PMID- 11426393 TI - Preparation and microstructure analysis of chitosan/hydroxyapatite nanocomposites. AB - Chitosan/hydroxyapatite (HAp) composites with a homogeneous nanostructure have been prepared by a co-precipitation method. According to TEM observations, HAp crystallites in the composites formed elliptic aggregations 230 nm in length and 50 nm in width. The typical length of the aggregations corresponded approximately to that of a chitosan molecule. The size of the constituent HAp crystallites was found to be predominantly 30 nm in length and 10 nm in width, and the c-axes were well aligned in parallel with the chitosan molecules in the respective aggregations. The growth of the HAp crystallites is considered to occur at nucleation sites, most probably forming the complexes with amino groups on chitosan with calcium ions. The compact composites obtained have been found to be mechanically flexible, and this flexibility has been improved further by heating at 120 degrees C for 20 min in an autoclave with saturated steam pressure. PMID- 11426394 TI - Repairing segmental bone defects with living porous ceramic cylinders: an experimental study in dog femora. AB - Twenty-one porous hydroxyapatite-tricalcium phosphate ceramic cylinders 1.5 cm long and 1.2 cm in diameter were swathed with fresh autogenetic periostea taken from 21 dogs and implanted in the same animals' muscles to get living ceramic bone substitutes. These substitutes contained autogenetic growth stimulators including osteoblasts and other multipotential cells. One month later, they were transferred to the segmental bone defect sites created in femoral diaphysis of the same animals. The roentgenograms showed that in time the boundaries between the cylinders and bone sections became vague. X-ray diffraction analysis indicated that the spectra of the samples tended to be similar to those of natural bones by 6 months postoperatively. Their bending strengths also increased gradually. After the cylinders were transferred from muscles to the bone defect sites, newly formed bone tissues rapidly increased and the cylinders gradually fused with the contacted bones 2 months later. By 4 and 6 months, bone tissue occupied most areas of the materials. The ratio of newly formed bone tissue had a large lead over the materials. Haversian systems were clearly observed and matured bone tissues filled the ceramic pores and connected with each other. Results suggested that culturing complexes that consist of autogenetic periostea and biomaterials, taking advantage of living organic culturing medium, should be an effective approach to get satisfactory bioactive bone substitutes. It also provides a basis for clinically repairing bone defects in bearing sites with complex bioceramics. PMID- 11426395 TI - In vitro biocompatibility assessment of naturally derived and synthetic biomaterials using normal human urothelial cells. AB - The reconstruction of urinary tissues often employs various types of biomaterials, and adequate material biocompatibility is essential for the successful reconstruction of urinary tissues. In this study we utilized a primary normal human urothelial cell culture system to evaluate the in vitro biocompatibility of a number of naturally derived biomaterials [i.e., bladder submucosa, small intestinal submucosa, collagen, and alginate] and polymeric biomaterials [i.e., poly(glycolic acid), poly(L-lactic acid), poly(lactic-co glycolic acid), and silicone] that are either experimentally or clinically used in urinary reconstructive surgery. To determine the cytotoxic and bioactive effects of these biomaterials, the cell viability, metabolic activity, apoptotic properties, and DNA-synthesis activity were measured with four types of assays [Neutral Red, 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyl tetrazolium bromide, apoptotic activity, and tritiated thymidine incorporation assays] using extract and direct contact methods. Most of the biomaterials tested did not induce significant cytotoxic effects and exhibited normal metabolic function and cell growth in vitro. This normal primary human urothelial cell culture model is suitable for in vitro biocompatibility assessments and is able to provide information on the cell-biomaterial interactions and the ability of biomaterials to support bioactive cell functions. PMID- 11426396 TI - Biomechanical examinations of cancellous bone concerning the influence of duration and temperature of cryopreservation. AB - The influence of cryopreservation of cancellous bone at different temperatures and different storage periods was investigated for mechanical changes. Standardized cancellous cylinders (50 x 25 mm) were stored at -20 degrees C or 80 degrees C for 6 or 12 months, respectively, or for 2 years and then subjected to a screw pull-out test. No significant differences were found among the various storage temperatures and periods investigated. We conclude that different cryopreservation temperatures do not have any influence on the mechanical properties of cancellous bone. Also, storage duration up to 2 years has no effects on the mechanical strength of the bone. PMID- 11426397 TI - Interactions between calcium, phosphate, and albumin on the surface of titanium. AB - The deposition of calcium phosphate on chemically polished commercially pure titanium immersed in Hank's balanced salt solution (HBSS) with bovine serum albumin (BSA) (concentrations 0 and 4 mg/mL) has been investigated. Electrochemical techniques, 125I labeling of albumin, scanning electron microscopy, energy dispersive spectroscopy, X-ray diffraction, and X-ray photoelectron spectroscopy were used. A tricalcium phosphate layer with a thickness of ca. 1 microm was formed for periods of immersion in HBSS ranging between 1 and 2 weeks. A concentration of 4 mg/mL of BSA prevented its formation, even for periods as long as 1 month. In the absence of BSA, the electrochemical behavior of titanium specimens was significantly affected by the length of immersion time, reflecting the changes that slowly occur on their surface. In the presence of BSA, the surfaces maintained most of their original electrochemical activity. Surface studies have shown that calcium and phosphate become incorporated in the surface at very early stages of immersion. Albumin, which was rapidly adsorbed on titanium, was slowly desorbed when titanium was placed in HBSS. Protein and phosphate may coexist on the same surface, but initially adsorbed albumin molecules prevent the precipitation of a thick layer of tricalcium phosphate. PMID- 11426398 TI - Effect of compression on fast swelling of poly(acrylamide-co-acrylic acid) superporous hydrogels. AB - Superporous hydrogels (SPHs) swell to a large size in a very short time. In many applications it is preferred to compress SPHs to reduce the overall dimension in the dried state. The effects of compression on the swelling property of SPHs were examined. The swelling property of the compressed SPHs was dependent on the orientation of the SPHs during compression. If SPHs were compressed in an orientated manner so that they retained interconnected porous structure, they were able to swell to near equilibrium within 10 min of immersion in aqueous fluids. If SPHs were compressed in a manner that did not retain the open pore structure, the swelling rate was greatly reduced. The results showed that the SPHs could be compressed without significant sacrifice of the fast swelling property if compressed in a proper orientation. Because pores were formed owing to the generation of gas which rose from bottom to the top of the container, the compression parallel to the pore formation resulted in preservation of the pore structure, and thus fast swelling property. The ability to compress SPHs, maintaining the fast swelling property, is expected to be useful in various applications including development of gastric retention devices for oral drug delivery. PMID- 11426399 TI - Mechanical regulation of localized and appositional bone formation around bone interfacing implants. AB - The local mechanical environment around bone-interfacing implants determines, in large part, whether bone formation leading to functional osseointegration will occur. Previous attempts to relate local peri-implant tissue strains to tissue formation have not accounted for implant surface geometry, which has been shown to influence early tissue healing in vivo. Furthermore, the process by which mechanically regulated peri-implant bone formation occurs has not been considered previously. In the current study, we used a unit cell approach and the finite element method to predict the local tissue strains around porous-surfaced and plasma-sprayed implants, and compared the predictions to patterns of bone formation reported in earlier in vivo experiments. Based on the finite element predictions, we determined that appositional bone formation occurred when the magnitudes of the strain components at the tissue-host bone interface were <8%. Localized, de novo bone formation occurred when the distortional tissue strains were less than approximately 3%. Based on these threshold tissue strains, we propose a mechanoregulatory model to relate local tissue strains to the process of peri-implant bone formation. The mechanoregulatory model is novel in that it predicts both appositional and localized bone formation and its predictions are dependent on implant surface geometry. The model provides initial criteria with which the osseointegration potential of bone-interfacing implants may be evaluated, particularly under conditions of immediate or early loading. PMID- 11426400 TI - Interaction of sulfonylurea-conjugated polymer with insulinoma cell line of MIN6 and its effect on insulin secretion. AB - A carboxylated derivative of sulfonylurea (SU), an insulinotropic agent, was synthesized and grafted onto a water-soluble polymer as a biospecific and stimulating polymer for insulin secretion. To evaluate the effect of the SU conjugated polymer on insulin secretion, its solution in dimethyl sulfoxide was added to the culture of insulinoma cell line of MIN6 cells to make 10 nM of SU units in the medium and incubated for 3 h at 37 degrees C. The culture medium was conditioned with glucose concentration of 3.3 or 25 mM. To verify the specific interaction between the SU (K+ channel closer)-conjugated polymer and MIN6 cells, the cells were pretreated with diazoxide, an agonist of adenosine triphosphate sensitive K+ channel (K+ channel opener), before adding the SU-conjugated polymer to the cell culture medium. This treatment suppressed the action of SUs on MIN6 cells. Fluorescence-labeled polymer with rodamine-B isothiocyanate was used to visualize the interactions, and we found that the labeled polymer strongly absorbed to MIN6 cells, probably owing to its specific interaction mediated by SU receptors on the cell membrane. The fluorescence intensity on the cells significantly increased with an increase in incubation time and polymer concentration. A confocal laser microscopic study further confirmed this interaction. The results from this study provided evidence that SU-conjugated copolymer stimulates insulin secretion by specific interactions of SU moieties in the polymer with MIN6 cells. PMID- 11426401 TI - Fibronectin modulates macrophage adhesion and FBGC formation: the role of RGD, PHSRN, and PRRARV domains. AB - To probe the role of human plasma fibronectin in modulating human blood-derived macrophage adhesion and fusion to form multinucleated foreign-body giant cells (FBGC), a series of biomimetic oligopeptides based on the functional structure of fibronectin was designed and synthesized. Peptides incorporated the RGD and PHSRN integrin-binding sequences from FIII-10 and FIII-9 modules, respectively, and the PRRARV sequence from the C-terminal heparin-binding domain, either alone or in combination. Peptides were immobilized onto a polyethyleneglycol-based polymer substrate. The following conclusions were reached. Fibronectin modulated macrophage adhesion and the extent (i.e., size) of FBGC formation on control surfaces in the presence of serum proteins. Macrophages adhered to all substrates with relatively subtle differences between adhesion mediated by RGD, PHSRN, PRRARV, or combinations thereof. beta1-integrin subunit was essential in macrophage adhesion to peptide-grafted networks in a receptor-peptide specific manner, whereas beta3-integrin subunit was less important. Macrophage adhesion to PRRARV was mediated primarily by the direct interaction with integrins. RGD or PHSRN alone did not provide an adequate substrate for macrophage fusion to form FBGCs. However, the PHSRN synergistic site and the RGD site in a single oligopeptide provided a substrate for FBGC formation that was statistically comparable to that on the positive control material in the presence of serum proteins. This response was highly dependent upon the relative orientation between RGD and PHSRN. PRRARV did not support FBGC formation. These results demonstrate the importance of fibronectin and, specifically, the synergy between RGD and PHSRN domains, in supporting macrophage fusion to form FBGCs. PMID- 11426402 TI - Effect of osteogenic protein 1/collagen composite combined with impacted allograft around hydroxyapatite-coated titanium alloy implants is moderate. AB - This study evaluated the effects of osteogenic protein 1/collagen composite (OP 1/col) mixed with impacted allograft around hydroxyapatite (HA)-coated titanium alloy implants in a canine model. The aim of the study was to test different doses of OP-1 growth factor in a collagen composite for stimulatory effect on allograft incorporation around an implant. Unloaded implants were inserted in each proximal humerus of 16 skeletally mature dogs. The cylindrical implants (4 x 9 mm) coated with HA were initially surrounded by a 3-mm gap into which allograft mixed with OP-1/col was impacted. Two different doses of OP-1 were investigated. In eight animals 325 mg OP-1 protein and 130 mg bovine collagen type I as carrier were mixed with the allograft chips. This composite is identical to the clinically used OP-1 device called Novus. In another eight animals a lower dose of 65 mg OP-1 protein and 130 mg bovine collagen type I was used. Control implants placed in the contralateral humerus were surrounded by allograft mixed with collagen carrier only. The dogs were euthanized at 6 weeks. Implant fixation was determined by push-out testing. Bone ingrowth and bone formation were evaluated by quantitative histomorphometry on serial sections of the bone-implant interface. Impacted allograft together with low-dose OP-1 enhanced bone volume in a zone adjacent to HA-coated titanium alloy implants. The high dose had no effect on bone formation. Mechanical fixation, bone ingrowth, and bone volume in the gap near the original trabecular bone were unaffected by both low and high OP-1/col composite. In this model and observation period, the low dose of OP-1/col composite mixed with impacted allograft has a moderate effect on bone healing around HA-coated implants and no effect on implant fixation. PMID- 11426403 TI - Controlled delivery of taxol from poly(ethylene glycol)-coated poly(lactic acid) microspheres. AB - The development of injectable microspheres for sustained drug delivery to the arterial wall is a major challenge. We demonstrated the possibility of entrapping an antiproliferative agent, taxol, in poly(ethylene glycol) (PEG)-coated biodegradable poly(lactic acid) (PLA) microspheres with a mean diameter of 2-6 microm. A solution of taxol and PLA dissolved in an acetone/dichloromethane mixture was poured into an aqueous solution of PEG [or poly(vinyl alcohol) (PVA] with stirring with a high-speed homogenizer for the formation of microspheres. Taxol recovery in PLA-PEG microspheres was higher (61.2 +/- 2.3%) than with PVA based (41.6 +/- 1.8%) preparations. An analysis by diffuse reflectance infrared Fourier transform spectroscopy revealed that PEG was incorporated well on the PLA microsphere surface. Scanning electron microscopy revealed that the PEG-coated PLA microspheres were spherical in shape and had a smooth surface texture like those of PVA-based preparations. The amount of drug release was much higher initially (25-30%); this was followed by a constant slow-release profile for a 30 day period of study. This PEG-coated PLA microsphere formulation may have potential for the targeted delivery of antiproliferative agents to treat restenosis. PMID- 11426404 TI - Web alert. PMID- 11426405 TI - Web alert. PMID- 11426406 TI - Neuromuscular disorders:gene location. PMID- 11426407 TI - Glucocorticoids and the thymus: the view from the middle of the road. PMID- 11426408 TI - [Prefrontal cortex in memory and attention processes]. AB - INTRODUCTION: The role of the prefrontal cortex still remains poorly understood. Only after 1970, the functions of the frontal lobes have been conceptualized from different points of view (behaviorism, cognitivism). DEVELOPMENT: Recently,different parallel circuits connecting discrete cortical and subcortical regions of the frontal lobes have been described. Three of these circuits are the most relevant to understanding of behavior: the dorsolateral prefrontal circuit, that mediates executive behavior; the orbitofrontal prefrontal circuit, mediating social behavior, and the medial frontal circuit, involved in motivation. Damage to the frontal cortex impairs planning, problem solving, reasoning, concept formation, temporal ordering of stimuli, estimation, attention, memory search, maintaining information in working memory, associative learning,certain forms of skilled motor activities, image generation and manipulation of the spatial properties of a stimulus, metacognitive thinking, and social cognition. Several theories have been proposed to explain the functions of the prefrontal cortex. Currently,the most influential cognitive models are: the Norman and Shallice supervisory attentional system, involved in non-routine selection; the Baddeley working memory model with the central executive as a supervisory controlling system, in which impairment leads to a 'dysexecutive syndrome'; and the Grafman's model of managerial knowledge units, stored as macrostructured information in the frontal cortex. CONCLUSION: The prefrontal cortex is essential for attentional control, manipulation of stored knowledge and modulation of complex actions, cognition, emotion and behavior. PMID- 11426409 TI - [Amusias]. AB - INTRODUCTION: We review the definition of the amusias and the cultural components involved in musical experience. We review the characteristics of musical language, perception and performance of it together with reading and writing it. DEVELOPMENT: We analyze the above data and significant clinical cases. We discuss whether in evaluation of the amusias it may be relevant to have had been taught music, since in this condition the functional specialization of the cerebral cortex has different topography of the lesions in persons who have or have not had musical education. CONCLUSIONS: We consider that in persons who have studied musical language, amusia should be evaluated as a second language, and its pathology as aphasia for musical language. In persons without musical education the dysperceptive amusias are a purely agnostic phenomenon with neuropsyschological characteristics which are different to those seen in persons who have had an education in music. PMID- 11426410 TI - [Attention processes and neuropsychological syndromes]. AB - INTRODUCTION: Both cognitive psychology and neuropsychology require an approach to the multidimensional 'attention'. DEVELOPMENT: Attention permits behaviour to be suitable to the task in hand. According to the situation or activity required, the components or mechanisms of attention necessary vary (selectivity, alertness, vigilance, focus, inhibitory control mechanisms, etc.)as do the different neural networks involved. In the different neuropsychological syndromes different components and attention mechanisms are affected. For instance, in attention deficit disorder there is inability to take notice of control of carrying out inappropriate responses, whilst in visual heminegligence there may be a defect of a mechanism such as disconnection of attention selective to a given space. To obtain coherent results in the study of attention disorders in various syndromes, the different components and attention mechanisms should be systematically evaluated using specific tests. In neuropsychology frequently the tests used only measured the aspects most closely connected to speed of perception and visual selective attention, without evaluating other components. Sometimes study data measuring different aspects of attention have been compared, and therefore the conclusions drawn regarding possible attention disorders were controversial. The lack of standardised data and the difficulty of establishing 'pure' tests of attention mechanisms make the task more arduous. CONCLUSION: Only the integration of experimental studies of cognitive psychology and the techniques and procedures of neuropsychology permit clarification of the construction and the advance of explanatory models. PMID- 11426411 TI - [Post-traumatic amnesia syndrome]. AB - INTRODUCTION: Mnesic deficits are frequent in subjects which have suffered from head injury and may persist during may years. OBJECTIVE: We will analyze posttraumatic amnesia characteristics (PTA) as well as learning and memory deficits which are normally observed once the PTA phase is over. DEVELOPMENT: The PTA is defined as the period that follows a brain injury in which the affected person is incapable of consistently remembering at least the last 24 hours. That is, the period after the head injury in which the incorporation of new information in long term memory is not possible. The study of PTA has generally been focussed on the analysis of the alterations of temporo-spatial orientation and mnesic deficits, however other cognitive and behavioral alterations do exist (linguistic, attentional, critical judgement, information processing, perception, etc.), and they are associated with the particular memory disorder that we are studying, given their influence on it. We will take on board the different theories that have been proposed to explain mnesic deficits which occur during the PTA phase: lack of consolidation of new information within longterm memory; recall deficit as a result of inefficient coding of information; failures of the mechanisms to carry out the process of consolidation-recall, and poor organization in the coding of new material. We will put forward a guide for the neuropsychological assessment of memory based on the analytic and concrete study of each mnesic cognitive component, sustained generally by specific neurophysiological functional systems which allow us to establish a diagnosis, prognosis, and adequate therapeutic focus for each concrete case. PMID- 11426412 TI - [Apraxia of ideas and movements and visual-constructive skills]. AB - OBJECTIVES: To study alterations in copying behaviour in Alzheimer's disease (AD) and its relationship with other cognitive parameters. As secondary objectives, in this paper we describe the evolution of concepts, definitions and examination of ideas-movement and constructive apraxia, together with its involvement in AD and review the investigations of the Uruguayan school on the development of behaviour copying and how this is affected in patients with cerebral lesions. PATIENTS AND METHODS: We made a study of 82 consecutive patients with probable AD at stages 3 to 5 on the overall deterioration scale (GDS), registered in the Department of Neuropsychology, and a group of healthy persons, paired for age, sex and educational level. We used an extensive protocol for cognitive evaluation. We evaluated the idea-movement apraxia by means of imitation of increasingly complex gestures, and constructive apraxia using the protocol described by Mendilaharsu et al. RESULTS: At stages 4 and 5 of the GDS scale, the idea-movement apraxia was significantly correlated with digital agnosia and the anomias. Constructive apraxia was found in GDS stage 3. We found 'homogeneous cognitive deterioration' in advanced patients (GDS 5) who showed closing-in when copying figures. The closing-in correlated significantly with the presence of anomias, idea apraxia and digital agnosia. CONCLUSION: In the early stages, EA shows varied praxis profiles, but above stage 5 of the GDS scale there was a tendency to homogeneity. PMID- 11426413 TI - [Attention systems and unilateral neglect]. AB - Unilateral neglect can be defined as an impairment to detect, refer, orient or respond to stimuli presented contralaterally to a cerebral lesion, without any impairments in sensory-motor elementary functions. Development. The first descriptions were those of Hughlings Jackson (1876) and Anton (1893). The most important feature of the syndrome is a lateralization bias, which is directional in nature independently of the visual fields. It can be classified in: 1. Attentional (sensory neglect). 2. Intentional (motor neglect). The lesions that may be responsible for the neglect syndromes are usually found in the inferior parietal cortex of the right cerebral hemisphere (superior parietal when optic ataxia is prominent), and the right frontal lobe. Extinction is more frequently related to subcortical lesions (right lenticular nucleus, anterior aspects of peri-ventricular white matter). Unilateral neglect comprises a set of features that may coexist or be isolated traits: 1. Attentional neglect: hemi-inattention, allesthesia, allochiria, anosognosia (with/without somato-paraphrenia or misoplegia), and anoso-diaphoria, sensory extinction. 2. Intentional neglect: hemi-akinesia, directional hypokinesia, motor impersistence, motor extinction. Different theories try to explain the patho-physiology of these phenomena: the attentional-intentional model, the neural network model, the vectorial model, the representational model, and the premotor model, among others. PMID- 11426414 TI - [Myasthemia gravis and myotonic dystrophy in the same patient]. PMID- 11426415 TI - [Tropical spastic paraparesis due to HTLV-1. A case report]. PMID- 11426416 TI - [Dubowitz syndrome, polymyositis and aleucemic myeloblastic leucemia: a new association]. PMID- 11426417 TI - Genomics: saviour or millstone? AB - The Keystone Symposium on the impact of Genomics Development was held in Santa Fe, New Mexico, from 2 to 7 February 2001. PMID- 11426418 TI - Somatic gene mosaicism and expressivity. PMID- 11426419 TI - CGRP receptors: beyond the CGRP(1)-CGRP(2) subdivision? PMID- 11426420 TI - Unified nomenclature for the subunits of eukaryotic initiation factor 3. PMID- 11426421 TI - [To the Editor: The problem of celiac disease in the present]. PMID- 11426422 TI - [To the Editor: Irregular position of the gallbladder]. PMID- 11426423 TI - Lid retraction in primary hepatocellular carcinoma. PMID- 11426424 TI - Bilateral birdshop retinochoroidopathy in absence of overt clinical lesions. PMID- 11426425 TI - Anterior ischaemic optic neuropathy in a patient with HLA-B27 associated anterior uveitis and ankylosing spondylitis. PMID- 11426426 TI - Cyclical central serous chorloretinopathy associated with cystoid macular oedema. PMID- 11426427 TI - Endogenous Rhodotorula minuta and Candida albicans endophthalmitis in an injecting drug user. PMID- 11426428 TI - The disappearing "melanoma". PMID- 11426429 TI - Binding and orientation of fibronectin on surfaces with collagen-related peptides. Student Research Award in the Masters Science Degree Candidate category, 27th annual meeting of the Society for Biomaterials, St. Paul, MN, April 24-29, 2001. AB - Although fibronectin (FN) has been used in a variety of in vitro studies to enhance cell and bacteria adhesion, relatively little is known about the molecular interactions of FN with surfaces, particularly the interactions that can control the binding, conformation, and functionality of FN on these surfaces. Even less is known about approaches needed to control binding, orientation, and functionality of FN bound on surfaces. To begin to fill this gap in our knowledge, we hypothesized that functional FN can be bound and specifically oriented on polystyrene surfaces with FN-specific collagen-related peptides (CRPs). We further hypothesized that monoclonal antibodies that react with specific epitopes on FN can be used to quantify both FN binding and orientation on these surfaces. On the basis of these hypotheses, we initiated a systematic investigation of the binding and orientation of FN on polystyrene surfaces with CRPs. To bind FN to surfaces, we used two different CRPs: CRP-I (TLQPVYEYMVGV) and CRP-II (TGLPVGVGYVVTVLT). The binding and orientation of the FN molecule to these immobilized CRPs was quantified with (125)I-FN and monoclonal antibodies. Monoclonal antibodies used for this study were reactive with specific regions of the FN molecule, that is, the amino (N) terminus (anti-N antibodies) and carboxyl (C) terminus (anti-C antibodies). The results of our studies demonstrated that although CRP-I and CRP-II could be bound directly to polystyrene, these directly immobilized CRPs failed to bind (125)I-FN . Thus, to facilitate FN binding to the CRPs, we used bovine serum albumin (BSA) as a spacer to physically elevate the CRPs away from the polystyrene surface. Thus, CRP-I and CRP-II were covalently linked to BSA via the N and C termini of each CRP (CRP-I-BSA and CRP-II-BSA). (125)I-CRP-BSAs were all found to bind to equivalent levels on polystyrene (1.60 2.60 microg/cm2). When CRP-BSAs were immobilized on polystyrene, they all successfully bound (125)I-FN in a range of 34-72 ng/cm2 (mean). Using monoclonal antibodies to FN to characterize the orientation of FN bound to the various CRP BSAs, we demonstrated that (1) FN consistently bound to either CRP-I-BSA or CRP II-BSA; (2) bound FN reacted significantly more with anti-C antibodies than with anti-N antibodies; and (3) the increased reactivity of bound FN to anti-C antibodies was consistent, whether FN was bound by CRP-I or CRP-II or the CRPs were bound to BSA by the C or N termini. These data demonstrated an enhanced binding of anti-C antibodies to immobilized CRP-BSA relative to anti-N antibodies. We interpreted the data to be the result of FN binding in an oriented fashion with N termini of FN bound tightly to the BSA-polystyrene surface. In this position, the C termini of FN are exposed and available for binding by the anti-C antibodies. Alternatively, in this orientation the N termini of the FN would not be available to bind the anti-N antibodies, thereby explaining the decreased reactivity of the CRP immobilized FN to the anti-N antibodies. These studies not only demonstrate the utility of peptides in binding and orienting large molecular weight proteins such as FN on surfaces but underscore the need for well-characterized reagents (e.g., monomeric/functional FN and antibodies) to specifically bind, orient, and characterize large molecular weight proteins immobilized on various surfaces. PMID- 11426430 TI - C. Everett Koop, MD: a lifetime of achievement. The 1999 recipient of the Pennsylvania Medical Society's Distinguished Service Award. PMID- 11426431 TI - Reduced prefrontal cognitive activation associated with aggression in schizophrenia. PMID- 11426432 TI - Brave new world of post-genomics! AB - The Royal Society Discussion Meeting on "Utilizing the Genome Sequence of Parasitic Protozoa" was held at the Royal Society in London, UK, 21-22 March 2001. PMID- 11426433 TI - Sulfa-drug resistance in Plasmodium falciparum. PMID- 11426434 TI - 50 years ago in the Journal of Pediatrics: Congenital tracheo-esophageal fistula without esophageal atresia. PMID- 11426435 TI - 50 years ago in the Journal of Pediatrics: The effect of allergy management on growth and development of allergic children. PMID- 11426436 TI - 50 years ago in the Journal of Pediatrics: Pertussis: clinical comparisons of the newer antibiotics. PMID- 11426437 TI - Long-term outcomes of the ileal pouch anal anastomosis: the association of bowel function and quality of life 5 years after surgery. AB - INTRODUCTION: Previous studies have reported that mean health related quality of life (HRQL) levels generally attain normalcy following construction of an ileal pouch anal anastomosis (IPAA). It appears inconsistent, however, that these normal HRQL levels are achieved while bowel function (BF) scores generally remain statistically worse than "normal" (e.g., 4-8 stools/day, possible anal leakage, diaper usage). To investigate this inconsistency, the current study attempts to determine if any statistical associations are present between HRQL and BF, specifically in the long term. Multivariate regression analyses are performed using each of 8 individual HRQL domains against the full model of BF characteristics. METHODS: All patients more than 5 years status post an ileal pouch anal anastomosis (IPAA) procedure for familial adenomatous polyposis (FAP) at a single institution were studied. FAP was chosen because patients are routinely asymptomatic preoperatively. BF (e.g., stool frequency, anal leakage) and HRQL (using the 8 health domains of the SF-36) were assessed by patient interview. Student's t tests and full model multivariate regression analyses were used to analyze associations between BF and HRQL. RESULTS: The sample included 25 patients (14 male). Mean age was 39 years, mean follow-up time was 11 years. Although mean scores for the 8 individual HRQL domains were not statistically different from the general United States population, regression analyses of the different domains did demonstrate significant associations with varying levels of BF. While controlling for age and gender, the analyses show that the physical function domain is improved with the ability to pass flatus independent of stool, and physical role and mental health domains are improved with decreased stool frequency. The social function domain is improved with increased stool retention time, while the perception of general health is improved with less diaper usage and less sexual dysfunction. CONCLUSIONS: This study shows that a statistically significant association between HRQL levels and BF is present. Of the numerous BF characteristics tested, five appear to be of greater importance with regard to certain HRQL domains. This finding may have clinical implications concerning pouch construction and surgical technique. Methodologically, this study demonstrates that merely using mean levels to describe HRQL may not elucidate meaningful relationships between important clinical outcomes, such as function and HRQL. PMID- 11426438 TI - The effects of restricted energy and fluid intake on simulated amateur boxing performance. AB - This study examined the effects of serial reductions in energy and fluid intake on two simulated boxing performances separated by 2 days recovery. Eight amateur boxers (age: 23.6 +/- 3.2 years; height 175 +/- 5 cm; body mass [BM] 73.3 +/- 8.3 kg [Mean +/- SD]) performed two simulated boxing bouts (BB) under normal (N trial) and restricted (R-trial) diets in a counterbalanced design over 5 days. The trials were separated by a 9-day period of normal dietary behavior (X-trial). BM was recorded on days 1, 3, and 5 of each trial. Simulated bouts of three, 3 min rounds with 1-min recovery were completed on days 3 (BB1) and 5 (BB2) of each 5-day trial. Punching force (N) was recorded from 8 sets of 7 punches by a purpose-built boxing ergometer. Heart rate (fC) was monitored continuously (PE3000 Polar Sports Tester, Kempele, Finland), and blood lactate (BLa) and glucose (BG) were determined 4-min post-performance (2300 StaPlus, YSI, Ohio). Energy and fluid intakes were significantly lower in the R-trial (p < .05). Body mass was maintained during the N-trial but fell 3% (p < .05) during the R-trial. There were no significant differences in end-of-bout fC or post-bout BG, but BLa was higher in the N- than the R-trial (p < .05). R-trial punching forces were 3.2% and 4.6% lower, respectively, compared to the corresponding N-trial bouts, but the differences did not reach statistical significance. These results suggest that energy and fluid restrictions in weight-governed sports do not always lead to a significant decrease in performance, but because of the small sample size and big variations in individual performances, these findings should be interpreted with care. PMID- 11426439 TI - Lessons learned in a European-Latin American collaboration for developing postgraduate education in public health. AB - BACKGROUND: New funding from the European Union (EU) allowed the collaborative development of postgraduate education in public health between five countries in South America and Europe. METHODS: A planning meeting, two workshops, a final meeting and the use of e-mail and faxes for discussing curricula were employed. RESULTS: One new master of public health in Argentina has begun, plans for new masters in Chile have been developed. A new collaborative module using the Internet was started and is now in its third year, new European plans for distance learning are being developed. Possibilities for collaborative research have been identified and those involved developed a broader horizon and international contacts. The planned grant from the EU was 72,040 Euros. Staff time spent administering the project was much greater than time spent discussing curricula. This is the only publication. CONCLUSION: Although the aims were achieved, the opportunity costs were great, as methods of assessing European research do not value such work highly and the administrative burden of small EU grants is high. Once a network is established greater efforts should be put into electronic communication, but European programmes currently do not fund such 'remote' support. PMID- 11426440 TI - Web alert. PMID- 11426441 TI - Dermatitis herpetiformis and cutaneous T-cell lymphoma. PMID- 11426442 TI - Erythema nodosum complicated by retrobulbar optic nerve neuritis. PMID- 11426443 TI - Scarring alopecia due to folliculitis decalvans in a patient with Darier's disease. PMID- 11426444 TI - Psoriasis vulgaris associated with Vogt-Koyanagi-Harada syndrome. PMID- 11426445 TI - Dissipation of potassium and proton gradients inhibits mitochondrial hyperpolarization and cytochrome c release during neural apoptosis. AB - Exposure of rat hippocampal neurons or human D283 medulloblastoma cells to the apoptosis-inducing kinase inhibitor staurosporine induced rapid cytochrome c release from mitochondria and activation of the executioner caspase-3. Measurements of cellular tetramethylrhodamine ethyl ester fluorescence and subsequent simulation of fluorescence changes based on Nernst calculations of fluorescence in the extracellular, cytoplasmic, and mitochondrial compartments revealed that the release of cytochrome c was preceded by mitochondrial hyperpolarization. Overexpression of the anti-apoptotic protein Bcl-xL, but not pharmacological blockade of outward potassium currents, inhibited staurosporine induced hyperpolarization and apoptosis. Dissipation of mitochondrial potassium and proton gradients by valinomycin or carbonyl cyanide p-trifluoromethoxy phenylhydrazone also potently inhibited staurosporine-induced hyperpolarization, cytochrome c release, and caspase activation. This effect was not attributable to changes in cellular ATP levels. Prolonged exposure to valinomycin induced significant matrix swelling, and per se also caused release of cytochrome c from mitochondria. In contrast to staurosporine, however, valinomycin-induced cytochrome c release and cell death were not associated with caspase-3 activation and insensitive to Bcl-xL overexpression. Our data suggest two distinct mechanisms for mitochondrial cytochrome c release: (1) active cytochrome c release associated with early mitochondrial hyperpolarization, leading to neuronal apoptosis, and (2) passive cytochrome c release secondary to mitochondrial depolarization and matrix swelling. PMID- 11426446 TI - Exclusion of growth factor gene mutations as a common cause of Sotos syndrome. AB - Sotos syndrome is characterized by somatic overgrowth, i.e., macrocephaly and tall stature. Because the cause and pathogenesis of Sotos syndrome remain unknown, we selected nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT-3) as possible genes mutated in Sotos syndrome. In seven patients with the classic phenotype, we excluded mutations in these growth factor genes. It is possible that these three genes are not involved in the cause of Sotos syndrome, or alternatively, mutations could not be identified in the small number of patients studied. PMID- 11426447 TI - Mother to son transmission of del(1) (q42.1q42.3). AB - Familial transmission of cytogenetically visible autosome deletions is rare in humans. We describe here a case of mother to son transmission of an interstitial deletion of the distal long arm of chromosome one, breakpoints q42.1q42.3. This is the smallest described deletion of this region to date. PMID- 11426448 TI - Molecular and cytogenetic characterization of a prenatally ascertained de novo (X;Y) translocation. PMID- 11426449 TI - Documenting nursing expertise in genetics: where are we going? PMID- 11426450 TI - Psychological impact of receiving a BRCA1/BRCA2 test result. AB - Mutation analysis for autosomal dominant hereditary breast/ovarian cancer genes (BRCA1/BRCA2) became an important technique for women at risk of carrying these mutations. Healthy female mutation carriers have a high lifetime risk for breast and/or ovarian cancer and may opt for frequent breast and ovary surveillance or prophylactic surgery (mastectomy and/or oophorectomy). Psychological distress was assessed in 78 healthy women at risk of having inherited a BRCA1/BRCA2 mutation opting for genetic testing and 56 partners several weeks prior to ("pre-test") and after ("post-test") learning about their DNA test result. Twenty-five women were found to be mutation carriers, and 53 were non-mutation carriers. One goal of the study was to identify individuals at risk for high distress in the weeks following disclosure of the test result. Interview transcripts were used to give a fuller picture of pre- and post-test distress. High post-test anxiety was reported by 20% of the mutation carrier women and by 35% of their partners. Eleven percent of women without the mutation and 13% of their partners reported high post-test anxiety levels. High post-test anxiety in women was significantly related to 1) a high level of pre-test anxiety and 2) being a mutation carrier. Women without a mutation who had a sister identified as a mutation carrier recently had higher post-test levels of depression than the other non-mutation carriers. It is suggested to consider seriously the need for psychological support in mutation carriers who had been anxious at pre-test already. For most non-mutation carriers, psychological follow-up might be of lesser importance, but those having a sister receiving an unfavorable test result should be informed about the possibility that they might not feel relief. PMID- 11426451 TI - Evidence of skewed X-chromosome inactivation in 47,XXY and 48,XXYY Klinefelter patients. AB - Klinefelter (47,XXY) syndrome occurs in approximately 1:800 male births and accounts for about 10-20% of males attending infertility clinics. Recent studies have shown no obvious phenotypic differences between subjects in which the extra X-chromosome is of paternal or maternal origin; however, a minority of Klinefelter patients are adversely affected clinically and intellectually to an exceptional level, and the underlying basis of this phenotypic variation is not known. We hypothesize that skewed X-inactivation and possibly parental origin of the X-chromosomes is involved. In this study, we determined parental origin and inactivation status of the X-chromosomes in 17 cytogenetically confirmed 47,XXY cases, two 48,XXYY cases and one mosaic 46,XY/47,XXY case. Eight highly polymorphic markers specific to the X-chromosome and the polymorphic human androgen-receptor (HUMARA) methylation assay were used to determine the parental origin and X-inactivation status of the X-chromosomes, respectively. Overall, 17 cases were fully informative, enabling parental origin to be assigned. In 59% of cases, both X-chromosomes were of maternal origin (Xm); in the remaining 41%, one X was of maternal (Xm) and one was of paternal origin (Xp). In 5 of 16 (31%) cases informative at the HUMARA locus, skewed X-inactivation was observed as defined by greater than 80% preferential inactivation involving one of the two X chromosomes. The two 48,XmXpYY cases both showed preferential paternal X chromosome (Xp) inactivation. Three 47,XmXmY cases also showed preferential inactivation in one of the two maternal X-chromosomes. These results suggest that skewed X-inactivation in Klinefelter (47,XXY and 48,XXYY) patients may be common and could explain the wide range of mental deficiency and phenotypic abnormalities observed in this disorder. Further studies are warranted to examine the role of X-inactivation and genetic imprinting in Klinefelter patients. PMID- 11426452 TI - Oncology nurses' knowledge, practice, and educational needs regarding cancer genetics. AB - This study evaluated oncology nurses' knowledge of cancer genetics and related topics, and identified current practice patterns and perceived educational needs in this area. A 54-item study questionnaire was mailed to a random sample of 1,200 Oncology Nursing Society (ONS) members and 75 members of the ONS-Cancer Genetics Special Interest Group; 656 (51%) of those eligible responded. After exclusions, we analyzed 573 responses. Most respondents were Caucasian, female, and worked in hospital or outpatient settings. Half were staff nurses and 8% specialized in cancer genetics. Respondents with higher levels of nursing education or with continuing education in cancer genetics, who worked in positions other than staff nurses, and whose primary practice area was cancer genetics had significantly higher mean scores overall on questions measuring knowledge of cancer genetics and related areas. Higher perceived educational needs to improve knowledge or practice related to cancer genetics at basic, intermediate or advanced levels were associated with all or some of the following variables: lower education; hospital/ outpatient or managed care/private practice settings; lack of continuing education in cancer genetics, and positions other than advanced practice nurses. Although nearly half of the respondents had received patient inquiries regarding cancer genetics, only 35% were aware of referral resources and 26% had made such referrals. These findings may be used to develop targeted educational approaches that prepare oncology nurses to incorporate cancer genetics into any level of practice. PMID- 11426453 TI - Assignment of PGL3 to chromosome 1 (q21-q23) in a family with autosomal dominant non-chromaffin paraganglioma. AB - We performed a whole genome scan in a family with maternally transmitted paraganglioma (PGL3). The family included five patients with histologically proven paraganglioma and one patient with imaging findings consistent with a paraganglioma. In addition, there were 33 clinically unaffected family members. Of these eight could be examined by magnetic resonance imaging. Our investigations indicate that PGL3 is located in 1q21-q23 for several reasons: 1) two-point linkage analysis yielded the highest LOD score of 2.25 at 1q21-q23 (marker D1S2675); 2) haplotype analysis was most consistent for 1q21-q23 markers; and 3) the locus was excluded from more than 97% of the genome using a total of 381 highly polymorphic markers. PMID- 11426454 TI - Longitudinal study of the carrier testing process for fragile X syndrome: perceptions and coping. AB - This paper reports the results of a longitudinal study of women at-risk to inherit the fragile X mutation. It addresses 1) how upsetting the women perceived their carrier information to be, 2) how serious a problem they perceive fragile X syndrome to be, and 3) descriptions of feelings about the carrier testing process. The study sample consisted of 42 women (20 carriers and 22 noncarriers). There were two measurement times (just prior to carrier testing and after learning actual carrier status). The measures used were a Fragile X Visual Analog Scale and a structured interview. At time 1, being at-risk was reported to be upsetting and fragile X syndrome was perceived to be a serious problem. For the women found to be carriers there was no change from time 1 to time 2 on any of the items. Significant change occurred in the non-carriers. They were significantly less upset at time 2 after receiving the results of their carrier test than at time 1. They also perceived fragile X syndrome to be a more serious problem than they did at time 1 and a more serious problem than the carriers at time 2. Themes found included concerns that carrier status for fragile X syndrome presented a barrier for having healthy biological children and concern for children's and grandchildren's adaptation to their own carrier status. Coping behaviors were activated to manage the emotions related to these concerns. The coping behaviors identified were minimization, acceptance of the possibility of being a carrier, a sense of being able to deal with the outcome of the carrier test, positive comparison, problem solving, and positive interpretation. PMID- 11426455 TI - Sib understanding of genetics and attitudes toward carrier testing for X-linked severe combined immunodeficiency. AB - X-linked severe combined immunodeficiency (XSCID) is the most common genetic form of SCID, a rare disease with profoundly impaired immunity. SCID was previously fatal but now can be treated by bone marrow transplantation. Mapping of XSCID in 1985 and identification of the disease gene, IL2RG, in 1993 made possible patient and carrier diagnosis. We assessed understanding of the genetics of XSCID in adult sibs recruited from families in which a proband had enrolled in our protocols and had attended an XSCID family workshop. Thirty-seven female and three male sibs completed a questionnaire and semistructured interview. Overall knowledge of genetics of XSCID was excellent. An overwhelming majority of participants (93%) believed that daughters should be tested for XSCID carrier status; 89% would prefer to have their own daughter tested prior to age 18 years (M = 9, median = 12), and 34% would test at birth. Moreover, 89% felt they would disclose carrier results to their daughter before adulthood (M = 12 years, median = 12); 51% would tell prior to adolescence. XSCID sibs were optimistic about medical science and assertive in their search for the latest information. Genetic information should be made available to families over time and should include discussion of reproductive risks for sons surviving with XSCID and daughters as they grow up. We recommend that genetic counseling for XSCID include children in age-appropriate discussions and that counselors help parents weigh benefits of early testing and disclosure versus the potential harm of loss of child autonomy. PMID- 11426456 TI - Family pictures: growing up with a brother with X-linked severe combined immunodeficiency. AB - Severe combined immunodeficiency (SCID) is a rare syndrome of profoundly impaired immunity, most often X-linked (XSCID). In past generations male infants with XSCID succumbed to infections during the first year of life, but prompt diagnosis and bone marrow transplantation currently make survival possible for over 80%. This treatment typically requires hospitalization for several months; thus, the burden on the family is considerable. We assessed the psychological impact on sibs of boys with XSCID. Forty adult sibs from families studied by J.M.P. were interviewed by J.H.F., and rating scales developed. The majority expressed distress over prolonged maternal absence during the affected child's hospitalization; 67% believed the mother had unsuccessfully mourned son(s) who died of XSCID. Half of the sibs reported that communication in the family about XSCID had been poor. Families with a spontaneous mutation were significantly more likely to report separation issues (P = 0.05), perhaps due to stronger maternal guilt. Family communication was significantly related to parental mourning (P = 0.001) and to survivor guilt (P = 0.05). Difficulties for daughters included desire to repair the mother's loss of her own child, as well as attempts to undo feelings of being flawed, by heightened wishes to bear a healthy son. In light of these findings we suggest: 1) bone marrow transplantation and the period of isolation places great stress on the family; parents need help balancing needs of well sibs with needs of the affected son; 2) parents need help with mourning the loss of a son so family secrets will not prevail; and 3) sibs, both bone marrow donors and non-donors, face psychological risks and need support. PMID- 11426457 TI - Methylation of HpaII site at the human DXS16 locus on Xp22 as an assay for abnormal patterns of X inactivation. AB - The highly polymorphic human DXS16 locus on Xp22 contains a BglII restriction fragment length polymorphism with 33% heterozygosity. We report that methylation of the HpaII site, 3.1 kb away from this restriction fragment length polymorphism, correlates with X-inactivation. The BglII polymorphism distinguishes between the maternal and paternal alleles, and HpaII digestion identifies their methylation status. The accuracy of this assay was tested on more than 30 control females and some patients with known patterns of X inactivation. The data obtained from this assay agree substantially with those obtained using the androgen receptor assay, which is widely used for detecting patterns of X-inactivation. This is the first marker on Xp22 found to be suitable for clonal analysis. Of additional significance is this marker's proximity to the pseudoautosomal boundary on the X chromosome and its potential use in identifying rare events occurring in this region, which lead to escape from normal X inactivation. PMID- 11426458 TI - Prevalence and clinical characteristics of celiac disease in Downs syndrome in a US study. AB - Celiac disease is an autoimmune gastrointestinal disorder characterized by mucosal atrophy of the jejunum on exposure to gluten, a protein found in grains. The purpose of our study was to determine the prevalence of celiac disease in children with Downs syndrome in a U.S.-based Caucasian population. The 97 Downs syndrome children were screened for celiac disease using serum IgA-anti endomysial antibody testing, which is highly specific and sensitive for the disorder. Children with titers greater than 1:5 (using the IgA endomysial antibody [EMA] test; EMA+) were considered affected. Ten children (10.3%) were EMA+. We examined their HLA DQA1 DQB1 genotype, karyotype, clinical characteristics, and the prevalence of celiac disease in their first-degree relatives. The nine available karyotypes were trisomy 21. Downs syndrome-specific mean height percentile was 64%+/-26% (range <5-99%) and weight percentile was 43%+/-28% (range 5-95%). Presence of diarrhea, constipation, vomiting, and abdominal pain was similar for children with and without celiac disease. Only bloating symptoms were significantly more frequent in those with celiac disease (EMA+). Seven of eight (88%) genotyped EMA+ children had the celiac disease associated high-risk HLA DQA1*0501 DQB1*0201 genotype as compared with 13/ 80 (16%) of EMA- children. Five of 48 (10%) first-degree relatives of the celiac disease (EMA+) children were EMA+. In conclusion, celiac disease, as diagnosed by positive endomysial antibody tests, has an increased prevalence in children with Downs syndrome in the U.S. as compared with the general population (1/250). Clinical and growth characteristics do not distinguish between children with and without celiac disease. Based on these observations, it is recommended that children with Downs syndrome be screened for celiac disease. PMID- 11426459 TI - Subtle radiographic findings of achondroplasia in patients with Crouzon syndrome with acanthosis nigricans due to an Ala391Glu substitution in FGFR3. AB - A unique type of craniofacial dysostosis, Crouzon syndrome with acanthosis nigricans (CAN), has been attributed to a specific substitution (Ala391Glu) in the fibroblast growth factor receptor 3 (FGFR3) gene. At birth, individuals with this disorder have craniosynostosis, ocular proptosis, midface hypoplasia, choanal atresia, hydrocephalus, and they experience the onset of acanthosis nigricans during childhood. We report three cases and compare the clinical characteristics of our cases with the previously reported cases of this disorder. Since the Ala391Glu substitution in FGFR3 is close to the substitutions in the transmembrane domain that result in achondroplasia, we carefully reviewed the skeletal findings in six patients. We identified subtle radiographic findings of achondroplasia in all six cases including narrow sacrosciatic notches, short vertebral bodies, lack of the normal increase in interpediculate distance from the upper lumbar vertebrae caudally, and broad, short metacarpals and phalanges. Even before acanthosis nigricans appears, the presence of choanal atresia and hydrocephalus in an individual with features of Crouzon syndrome should suggest the diagnosis of CAN, and subtle skeletal findings can lend further support to this diagnosis. PMID- 11426460 TI - Manifestations in four males with and an obligate carrier of the Lenz microphthalmia syndrome. AB - Lenz microphthalmia syndrome is a rare X-linked recessive condition first described by Lenz in 1955 and comprises of anophthalmia, microcephaly, mental retardation, external ear, digital, cardiac, skeletal, and urogenital anomalies. We present three brothers (ages 15 years, 9 years, and 18 months) and a maternal uncle (age 27 years) with congenital anophthalmia, delayed motor development, hypotonia, and moderate to severe mental retardation. They also have abnormally modeled ears, high-arched palate, pectus excavatum, finger and toe syndactyly, clinodactyly, fetal pads, scoliosis, cardiac, and renal abnormalities. An obligate carrier had abnormally modeled ears and syndactyly of the 2nd to 3rd toes bilaterally. Linkage and haplotype analysis in this family indicates that the gene is located in a 17.65-cM region on chromosome region Xq27-Xq28. PMID- 11426461 TI - Update on the use of nonhuman primate models for preclinical testing of gene therapy approaches targeting hematopoietic cells. AB - Transfer of genes into hematopoietic stem cells or primary lymphocytes has been a primary focus of the gene therapy field for more than a decade because of the wide variety of congenital and acquired diseases that potentially could be cured by successful gene transfer into these cell populations. However, despite success in murine models and in vitro, progress has been slow, and early clinical trials were disappointing due to inefficient gene transfer into long-term repopulating cells. The unique predictive value of nonhuman primate or other large animal models has become more apparent, and major advances in gene transfer efficiency have been made by utilizing these powerful but expensive and complex systems. This review summarizes more recent findings from nonhuman primate investigations focusing on hematopoietic stem cells or lymphocytes as target populations, and highlights specific preclinical issues, including safety. Results from studies using standard retroviral vectors, lentiviral vectors, adenoviral vectors, and adeno-associated viral vectors are discussed. Judicious application of these models should continue to be a priority, and advances should now be tested in proof-of-concept clinical trials. PMID- 11426462 TI - Flow cytometric characterization of myogenic cell populations obtained via the preplate technique: potential for rapid isolation of muscle-derived stem cells. AB - Myoblast transplantation has been investigated as a therapy for muscle-related diseases and as a gene delivery vehicle for therapeutic recombinant proteins. Clinical successes involving muscle cell transplantation have been limited, in part because of poor donor cell survival, and the heterogeneous nature of myogenic donor cells has largely been ignored. We have previously reported an isolation technique, preplating, that results in purified myogenic cells that are capable of significantly higher rates of donor cell survival leading to enhanced gene transfer to skeletal muscle. Characterization of these purified cells revealed that they display markers common to stem cells and are capable of multilineage differentiation. This study was performed to phenotypically characterize, by flow cytometry, muscle-derived cell populations obtained by the preplate technique for the purpose of eventually developing a method to quickly identify and isolate viable muscle cells best suited for transplantation. Muscle cell cultures were analyzed for expression of the surface proteins Sca-1, c-Kit, and CD34. We found that the preplate technique purifies distinct myogenic cell subpopulations expressing CD34 alone (Sca-1 negative) and Sca-1 alone (CD34 negative), but that this expression is subject to change with time in culture. Isolation and transplantation of phenotypically pure Sca-1-positive myogenic cells, obtained by magnetic cell sorting, demonstrates the ability to quickly select viable myogenic cells capable of regenerating skeletal muscle and restoring dystrophin expression within dystrophic host skeletal muscle. Flow cytometric described phenotypes will aid in the rapid isolation of specific donor cell populations for muscle cell transplants and muscle cell-mediated gene therapies, thereby enhancing their future success. PMID- 11426463 TI - In vivo and in vitro correction of the mdx dystrophin gene nonsense mutation by short-fragment homologous replacement. AB - Targeted genetic correction of mutations in cells is a potential strategy for treating human conditions that involve nonsense, missense, and transcriptional splice junction mutations. One method of targeted gene repair, single-stranded short-fragment homologous replacement (ssSFHR), has been successful in repairing the common deltaF508 3-bp microdeletion at the cystic fibrosis transmembrane conductance regulator (CFTR) locus in 1% of airway epithelial cells in culture. This study investigates in vitro and in vivo application of a double-stranded method variant of SFHR gene repair to the mdx mouse model of Duchenne muscular dystrophy (DMD). A 603-bp wild-type PCR product was used to repair the exon 23 C to-T mdx nonsense transition at the Xp21.1 dys locus in cultured myoblasts and in tibialis anterior (TA) from male mdx mice. Multiple transfection and variation of lipofection reagent both improved in vitro SFHR efficiency, with successful conversion of mdx to wild-type nucleotide at the dys locus achieved in 15 to 20% of cultured loci and in 0.0005 to 0.1% of TA. The genetic correction of mdx myoblasts was shown to persist for up to 28 days in culture and for at least 3 weeks in TA. While a high frequency of in vitro gene repair was observed, the lipofection used here appeared to have adverse effects on subsequent cell viability and corrected cells did not express dystrophin transcript. With further improvements to in vitro and in vivo gene repair efficiencies, SFHR may find some application in DMD and other genetic neuromuscular disorders in humans. PMID- 11426464 TI - System for efficient helper-dependent minimal adenovirus construction and rescue. AB - Helper-dependent minimal adenoviral vectors deleted for all viral coding sequences are promising vectors for gene therapy. They retain only the adenovirus cis elements for replication and packaging, can accommodate up to 36 kb of foreign DNA, and exhibit prolonged transgene expression and reduced tissue toxicity as compared with first-generation adenoviral vectors. We have developed a system consisting of a set of cosmid cloning vectors (pMV and pMVX) for simple routine construction and efficient rescue of minimal adenoviral vectors. In the cloning vectors the inverted terminal repeats (ITRs) are flanked by recognition sites for the super rare-cutting endonuclease I-SceI. This allows the release of linear minimal adenovirus genomes for rescue of minimal adenovirus regardless of the sequence of the insert DNA. pMV contains a multiple cloning site for the insertion of 26 to 36 kb of therapeutic DNA. pMVX contains a noncoding human X chromosomal DNA fragment as a vector backbone, which provides endonuclease restriction sites that allow for complete or partial replacement of the vector backbone by 1 to 26 kb of therapeutic DNA sequences, while retaining a packageable final minimal adenovirus genome size between 27 and 37.5 kb. Both vectors exist in two forms, with or without an Escherichia coli lacZ reporter gene cassette. Several minimal adenoviral vectors with insert sizes ranging from 1.5 to 16 kb were constructed with these cloning vectors. Minimal adenoviruses were efficiently rescued and amplified to high titers, using a Cre/lox-based helper system. Vectors containing the X-chromosomal backbone were stable during amplification. This simple and efficient system facilitates the construction of minimal adenoviruses and should be useful for further improvement of these new vectors. PMID- 11426465 TI - Autologous antileukemic immune response induced by chronic lymphocytic leukemia B cells expressing the CD40 ligand and interleukin 2 transgenes. AB - Although the B cells of chronic lymphocytic leukemia (B-CLL cells) express both tumor-specific peptides and major histocompatibility complex (MHC) class I antigens, they lack the capacity for costimulatory signaling, contributing to their protection against host antitumor immunity. To stimulate CLL-specific immune responses, we sought to transfer the human CD40 ligand (hCD40L) gene to B CLL cells, using an adenoviral vector, in order to upregulate costimulating factors on these cells. Because efficient gene transduction with adenoviral vectors requires the expression of virus receptors on target cells, including the coxsackievirus B-adenovirus receptors (CAR) and alpha(v) integrins, we cocultured B-CLL cells with human embryonic lung fibroblasts (MRC-5 line). This exposure led to increased expression of integrin alpha(v)beta3 on B-CLL cells, which correlated with higher transduction rates. Using this novel prestimulation system, we transduced B-CLL cells with the hCD40L gene. The Ad-hCD40L-infected cells had higher expression of B7 molecules and induced activation of autologous T cells in vitro, but these T cells could not recognize parental leukemic cells. By contrast, an admixture of Ad-hCD40L-positive cells and leukemic cells transduced with the human interleukin 2 (IL-2) gene produced greater T cell activation than did either immunostimulator population alone. Importantly, this combination generated autologous T cells capable of specifically recognizing parental B-CLL cells. These findings suggest that the combined use of genetically modified CD40L-expressing B-CLL cells in combination with IL-2-expressing B-CLL cells may induce therapeutically significant leukemia-specific immune responses. PMID- 11426466 TI - Interleukin 12 gene therapy of cancer by peritumoral injection of transduced autologous fibroblasts: outcome of a phase I study. AB - A phase I dose-escalation clinical trial of peritumoral injections of interleukin 12 (IL-12)-transduced autologous fibroblasts was performed in patients with disseminated cancer for whom effective treatment does not exist. The goals of this study were to assess the safety and toxicities as well as the efficacy, and ancillarily the immunomodulatory effects, of peritumoral IL-12 gene transfer. Primary dermal fibroblasts cultured from the patients were transduced with retroviral vector carrying human IL-12 genes (p35 and p40) as well as the neomycin phosphotransferase gene (TFG-hIL-12-Neo). Patients received four injections at intervals of 7 days. Nine patients were enrolled in this dose escalation study, with secreted IL-12 doses ranging from 300 ng/24 hr for the first three patients to 1000, 3000, and 5000 ng/24 hr for two patients in each subsequent dosage level. Although a definite statement cannot be made, there appears to be perturbation of systemic immunity. Also, the locoregional effects mediated by tumor necrosis factor alpha (TNF-alpha) and CD8+ T cells were observed with tumor regression. Treatment-related adverse events were limited to mild to moderate pain at the injection site; clinically significant toxicities were not encountered. Transient but clear reductions of tumor sizes were observed at the injected sites in four of nine cases, and at noninjected distant sites in one melanoma patient. Hemorrhagic necrosis of tumors was observed in two melanoma patients. These data indicate that gene therapy by peritumoral injection of IL-12 producing autologous fibroblasts is feasible, and promising in patients with advanced cancer. PMID- 11426467 TI - Contribution of plasmid DNA to hepatotoxicity after systemic administration of lipoplexes. AB - Several studies have demonstrated that intravenous administration of DNA complexed with cationic lipid vectors induces the production of large quantities of proinflammatory cytokines. In this study we confirm these observations, using cationic lipid DOTAP and cationic phospholipid compounds. Moreover, we demonstrate that although intravenous administration of lipid-DNA complexes does not induce toxic effects in the lung, high transgene expression in lung correlates with histopathological lesions in liver, this fact being documented by high transaminase levels in serum of treated mice. We examine the contribution of various components of the lipoplexes in this observed liver toxicity, as well as in the increasing level of transaminases, and more particularly the role of nonmethylated CpG sequences of plasmid DNA. We show that blood samples from animals treated either with cationic lipid alone, or with cationic lipid complexed with methylated plasmid DNA, contain low levels of transaminases. The significant decrease in transaminase levels after injection of cationic lipid methylated pDNA complexes leads us to believe that nonmethylated CpG sequences could play a major role in this hepatoxicity. Similar results were observed when using a vector that did not encode a transgene, demonstrating that the expression of luciferase in lung was not responsible for this liver toxicity. All these observations suggest that significant work should be devoted to understand more clearly the mechanism of cationic lipid-DNA complex toxicity, and to overcome the problems subsequent to administration of non-methylated CpG sequences of plasmid DNA. PMID- 11426468 TI - Morphologic analysis of BMP-9 gene therapy-induced osteogenesis. AB - The present study was performed to determine the histological, ultrastructural, and radiographic changes that occur over time at intramuscular BMP-9 gene therapy treatment sites. Several members of the bone morphogenetic protein (BMP) family have the potential to induce osteochondrogenesis when the protein is delivered to rodents, canines, rabbits, and nonhuman primates. Previous studies have also demonstrated that BMP gene therapy utilizing adenoviral vectors can also stimulate orthotopic and heterotopic bone formation in rodents and rabbits. Athymic nude and Sprague-Dawley rats were injected with Ad-BMP-9 or Ad-beta-Gal (3.75 x 10(9) particles) in their thigh musculature and light microscopic, electron microscopic, and computerized tomography analysis was performed 3, 6, 9, 12, 15, 18, 21, and 100 days later. To assess early mesenchymal cell proliferation, a bromodeoxyuridine (BrdU) immunohistochemical analysis was also performed 48, 60, and 72 hr postinjection in athymic nude rats. All animals demonstrated extensive endochondral bone formation at the Ad-BMP-9 treatment sites within 3 weeks. The Sprague-Dawley rats also exhibited a massive, acute inflammatory infiltrate during the first week. Proliferating mesenchymal stem cells were clearly evident as early as 2 days after treatment, which differentiated into small or hypertrophied chondrocytes during the next week. During the third week, the cartilaginous matrix mineralized and formed woven bone, which converted to lamellar bone by 3 months. No evidence of bone formation was demonstrated at the Ad-beta-Gal injection sites in the athymic nude or Sprague-Dawley rats. In addition, no cellular proliferation was seen at the Ad beta-Gal treatment sites in the athymic nude animals as assessed by light microscopy and BrdU immunohistochemistry. The extensive bone formation induced by Ad-BMP-9 suggests that BMP gene therapy may have potential utility in the treatment of degenerative, rheumatic, or traumatic bone pathology. PMID- 11426469 TI - Health Department of the United Kingdom Gene Therapy Advisory Committee. Guidance on making proposals to conduct gene therapy research on human subjects. PMID- 11426470 TI - Department of Health and Human Services. Recombinant DNA Advisory Committee. Minutes of meeting June 28-29, 2000. PMID- 11426471 TI - Redefining in-hospital resuscitation: the concept of the medical emergency team. PMID- 11426472 TI - Evaluation of a hospital-wide resuscitation team: does it increase survival for in-hospital cardiopulmonary arrest? AB - OBJECTIVE: To assess the impact (defined not only with regard to patient outcome but also to record keeping for evaluation of care) of a formal, structured resuscitation team for in-hospital cardiopulmonary resuscitation over the year following its creation. METHODS: This is a "before and after" study in which charts of all patients needing resuscitation during the two-year period were reviewed and data arranged in the Utstein Style of in-hospital reporting of cardiac arrests. The review was limited to adults (> or = 18 years of age) in nonICU settings. RESULTS: A total of 220 events were identified. Demographics and presenting rhythms for the two periods under review were similar. For the period of August 1996-August 1997 (group 1), there were 70 resuscitation events recorded with a return of spontaneous circulation (ROSC) rate of 21/70 (30%). For the period of August 1997-August 1998 (group 2), 150 events were recorded and the ROSC rate was significantly higher 87/150 (58%)) (P=0.0002). ROSC after ventricular fibrillation and ventricular tachycardia was similar in both groups (50 vs 57%) (P = 1.00) but an improvement in survival was seen in group 2 from events of bradycardia perfusing rhythm (25% vs 84%) (P = 0.0003). Survival from PEA/Asystole was also improved during period 2 (18 vs 48%) (P = 0.013). Survival to discharge was seen in 3/50 (6%) of patients in period 1 and 18/102 (18%) of patients in period 2 (P = 0.09). CONCLUSIONS: The formation of a structured, formalized hospital resuscitation team was associated with an increase in the number of recorded events, in the number of patients experiencing ROSC and in the percentage of patients who were discharged from the hospital. Facilities with no formal resuscitation team or with no skilled, practiced resuscitator on their current team should consider implementation of a similar strategy. PMID- 11426473 TI - Factors associated with the occurrence of cardiac arrest during hospitalization for acute myocardial infarction in the second national registry of myocardial infarction in the US. AB - Cardiac arrest can occur as a complication of acute myocardial infarction (AMI). To date, few studies have described factors associated with cardiac arrest occurrence and survival during hospitalization for treatment of AMI. We used data from a large national registry of hospitalized AMI patients to identify these factors. Data were collected from 1073 participating institutions, representing 14.4% of US hospitals. Hospital site coordinators conducted periodic chart reviews for AMI patients and data were submitted to an independent center for periodic review. Univariate analysis and multivariate logistic regression were used to identify factors associated with cardiac arrest. We found that cardiac arrest occurred in 4.8% (14,725/305,812) of hospitalized AMI patients. The survival rate to hospital discharge for these individuals was 29.4%. Sustained ventricular tachycardia or fibrillation (VT/VF) was present in 34.7% and was associated with a higher rate of survival to hospital discharge compared to cardiac arrest patients without a ventricular tachyarrhythmia (47.5 vs. 19.8%, P < 0.00001). Hypotension (initial systolic BP < 90 mmHg), q-wave AMI, old age, heart failure and initial heart rate abnormalities (bradycardia or tachycardia) were associated with a higher prevalence of cardiac arrest. A higher percentage of women compared to men experienced cardiac arrest (6.0 vs. 4.41%, P < 0.0001). Cardiac arrest prevalence was lower in patients with inferior wall infarction than in other types of ST-elevation infarction. Use of reperfusion therapy (PTCA or tPA) was associated with improved survival compared to hospitalized AMI patients who did not receive such therapy. PMID- 11426474 TI - Characteristics and outcome among patients suffering in-hospital cardiac arrest in monitored and non-monitored areas. AB - AIM: To describe the characteristics and outcome among patients suffering in hospital cardiac arrest in relation to whether the arrest took place in a ward with monitoring facilities. METHODS: All patients who suffered an in-hospital cardiac arrest during a 4-year period in Sahlgrenska Hospital, Goteborg, Sweden and in whom resuscitative efforts were attempted, were prospectively recorded and described in terms of characteristics and outcome. RESULTS: Among 557 patients, 292 (53%) had a cardiac arrest in wards with monitoring facilities. Those in a monitored location more frequently had a confirmed or possible acute myocardial infarction (AMI) as judged to be the cause of arrest (P < 0.0001), and the arrest was witnessed more frequently (96 vs. 79%; P < 0.0001). Ventricular fibrillation/tachycardia was observed more often as initial arrhythmia in monitored wards (56 vs. 44%; P = 0.006). The median interval between collapse and first defibrillation was 1 min in monitored wards and 5 min in non-monitored wards (P < 0.0001). Among patients with arrest in monitored wards 43.2% were discharged alive compared with 31.1% of patients in non-monitored wards (P = 0.004). Cerebral performance category (CPC-score) at discharge was somewhat better among survivors in monitored wards. CONCLUSION: In a Swedish University Hospital 47% of in-hospital cardiac arrests in which resuscitation was attempted took place in wards without monitoring facilities. These patients differed markedly from those having arrest in wards with monitoring facilities in terms of characteristics, interval to defibrillation and outcome. A shortening of the interval between collapse and defibrillation in these patients might increase survival even further. PMID- 11426475 TI - Geographical distribution of cardiac arrest in Nottinghamshire. AB - BACKGROUND: to analyse the incidence of out-of-hospital cardiac arrest in Nottinghamshire; to ascertain its geographical distribution; and to determine whether the geography of coronary heart disease mortality and out-of-hospital cardiac arrest are the same. METHODS AND RESULTS: population based, retrospective study in the County of Nottinghamshire with a total population of 993,914 in an area of 2183 km2 divided into 191 electoral areas. In the 4 years from 1 January, 1991 to 31 December, 1994, 1634 patients sustained a cardiac arrest attributed to a cardiac cause (International Classification of Diseases codes 390-414 and 420 429) and were attended by the Nottinghamshire Ambulance Service. The overall crude mean incidence rate of community cardiac arrest per electoral area was 40.2 per 100,000 population (range 0-121.2). Thirteen electoral areas, relatively deprived according to the Townsend score, had a significantly greater than expected incidence rate of cardiac arrest (median of 75.6/100,000 per electoral area; interquartile range (IQR) 65.3, 83.8). Twelve relatively affluent electoral areas had a significantly lower than expected incidence rate (median of 18.5/100,000 per area (IQR 13.0, 28.7). After adjusting for deprivation index, there were no differences in coronary heart disease (CHD) mortality and community cardiac arrest in urban and rural electoral areas. Apart from response times by ambulance crews, the events that follow the cardiac arrest such as bystander resuscitation, ventricular fibrillation found as the presenting rhythm and survival were similar in all electoral areas. CONCLUSIONS: increasing level of deprivation is associated with areas of increased incidence of out-of-hospital cardiac arrest in Nottinghamshire, and the effect is apparently different from that on CHD mortality. There is scope for reducing incidence rates of community cardiac arrest and to introduce strategies to improve survival in areas identified as having high rates of community cardiac arrest. PMID- 11426476 TI - A qualitative and quantitative study of the incidence, features and aetiology of near death experiences in cardiac arrest survivors. AB - AIM: To carry out a prospective study of cardiac arrest survivors to understand the qualitative features as well as incidence, and possible aetiology of near death experiences (NDEs) in this group of patients. METHOD: All survivors of cardiac arrests during a 1 year period were interviewed within a week of their arrest, regarding memories of their unconscious period. Reported memories were assessed by the Greyson NDE Scale. The postulated role of physiological, psychological and transcendental factors were studied. Physiological parameters such as oxygen status were extracted from the medical notes. Patients' religious convictions were documented in the interviews and hidden targets were used to test the transcendental theories on potential out of body claims. Those with memories were compared to those without memories. RESULTS: 11.1% of 63 survivors reported memories. The majority had NDE features. There appeared to be no differences on all physiological measured parameters apart from partial pressure of oxygen during the arrest which was higher in the NDE group. CONCLUSIONS: Memories are rare after resuscitation from cardiac arrest. The majority of those that are reported have features of NDE and are pleasant. The occurrence of NDE during cardiac arrest raises questions about the possible relationship between the mind and the brain. Further large-scale studies are needed to understand the aetiology and true significance of NDE. PMID- 11426477 TI - How often should defibrillation pads be changed?: the effect of evaporative drying. AB - OBJECTIVE: In order to minimise transthoracic impedance (TTI) during defibrillation, water-based pads are used to improve conductivity between metal defibrillation paddles and skin. Subjectively, these pads appear to dry very quickly; an effect that may lead to an increase in TTI due to poorer conduction between paddles and skin. This study was carried out to assess the effect of evaporative drying of defibrillation pads on TTI. MATERIALS AND METHODS: TTI was measured at 5-10 min intervals in 20 adult male volunteers across 3M defibrillation pads (2346N) placed in the anterior-apical position. Measurements of TTI were made at 30 kHz using a Bodystat MultiScan 5000 monitor at end expiration. A third pad was placed on the left precordium and its mass recorded each time a TTI measurement was made. RESULTS: The median age of subjects was 22 years (range 21-52 years) and their median body mass index was 23.1 kg m(-2) (range 18.4-42.8 kg m(-2)). Median room temperature was 23.0 degrees C (range 19.0-24.0 degrees C) and the median humidity was 31.0% (range 28.0-48.0%). 3M defibrillation pads had an initial mean mass of 25.14 g (S.D. +/- 0.41 g). Changes in defibrillation pad mass due to evaporative loss occurred immediately and rapidly, with all measurements being significantly lower than the baseline value. Mean baseline TTI was 63.6 ohms (S.D. +/- 13.7 ohms). After 30 min a statistically significant (P = 0.012) rise of 1.4 ohms (2.2%), was observed corresponding to a 12.6% decrease in pad mass, after which TTI continued to increase in a linear fashion. CONCLUSION: In the absence of a defibrillation current. 3M defibrillation pads can safely be left on the chest wall for at least 25 min in a typical hospital environment before evaporative drying results in a significant increase in transthoracic impedance. PMID- 11426478 TI - Long-term mild hypothermia with extracorporeal lung and heart assist improves survival from prolonged cardiac arrest in dogs. AB - BACKGROUND AND PURPOSE: although normothermic extracorporeal lung and heart assist (ECLHA) improves cardiac outcomes, patients can not benefit from hypothermia-mediated brain protection. The present study evaluated the effects of long-term ECLHA with mild to moderate hypothermia (33 degrees C) in a canine model of prolonged cardiac arrest. METHODS: 15 dogs were assigned to either the hypothermic (seven dogs, 33 degrees C) or normothermic group (eight dogs, 37.5 degrees C). All dogs were induced to normothermic ventricular fibrillation (VF) for 15 min, followed by 24 h of ECLHA and 72 h of intensive care. The hypothermia group maintained core (pulmonary artery) temperature at 33 degrees C for 20 h starting from resuscitation, then were rewarmed by 28 h. Outcome evaluations included: (1) mortality; (2) catecholamine dose; (3) time to extubation; (4) necrotic myocardial mass (g); and (5) neurological deficits score (NDS). RESULTS: in the normothermic group five dogs died of cardiogenic shock and one dog succumbed to poor oxygenation. The two surviving dogs remained comatose (NDS 60.5 +/- 4.9%) with necrotic myocardial mass of 14.5 +/- 3.5 g. In the hypothermic group, one dog died from pulmonary dysfunction, the other six dogs survived. The surviving dogs showed brain damage (29.8 +/- 2.5%), but there was evidence of some brain-protective effect. The mass of necrotic myocardium was 4.2 +/- 1.3 g in the hypothermic group or 3.4 times smaller than in the normothermic group. The survival rate was significantly higher in the hypothermic than in the normothermic group (P < 0.05). The catecholamine requirement was also lower in the hypothermic than in the normothermic dogs (P < 0.05). CONCLUSIONS: Long-term mild to moderate hypothermia with ECLHA induced immediately after cardiac arrest improved survival as well as cerebral and cardiac outcomes. PMID- 11426479 TI - An exceptional case of complete neurologic recovery after more than 5-h cardiac arrest. AB - We describe a case of more than 5 h cardiac arrest in a 60-year-old patient who underwent general anesthesia for a urologic operation. Before extubation, the patient suddenly developed ventricular fibrillation, pulseless ventricular tachycardia and asystole which was immediately treated by advanced life support (ALS) measures. Thirty minutes later seizures developed and were controlled by 200 mg of thiopentone and 10 mg of diazepam. A pattern of ventricular tachycardia, coarse ventricular fibrillation and asystole lasted for nearly 120 min. Termination of resuscitation maneuvers was considered, but long-term life support was continued for 5 h. After this time, peripheral pulses, with a supraventricular tachycardia-like rhythm and regular spontaneous breathing reappeared. Seven hours later, the patient had a Glasgow Coma Scale (GCS) of 5, dilated unresponsive, absence of pupils, and a systolic arterial pressure of 100 mmHg. He was then transferred to intensive care unit (ICU). The morning after, the patient was awake, responded to simple orders, breathing spontaneously, and free from sensomotor deficit. He was, therefore, extubated. Subsequently, other episodes of transitory ST-line upper wave followed by ventricular fibrillation appeared, suggesting Prinzmetal angina. This was successfully treated by percutaneous coronary angioplasty. The first electroencephalogram recorded the day after cardiac arrest showed a mild widespread background slowing. An electroencephalogram 6 days later showed a return to alpha rhythm with only mild theta-wave abnormalities. Four weeks after the first cardiac arrest the patient was discharged. This is an exceptional experience compared with the others reported. We believe that all the efforts must not be given up when such an event occurs during anesthesia and there are optimal conditions for resuscitation maneuvers. PMID- 11426480 TI - Failure of mouth-to-mouth resuscitation in cases of sudden infant death. AB - We describe two cases of sudden infant death syndrome (SIDS) and one case of apparent life threatening apnoea where resuscitation was attempted by the mouth to-mouth route. This was associated with evidence of gastric distension, including reflux of milk into the airway in the first two cases. In the second case the mother used mouth-to-mouth breathing after finding that she could not cover her baby's nose-and-open-mouth with her mouth. In the last case, the mother went on to try the mouth-to-nose route, with a good outcome. Systematic documentation of the route of resuscitation and its outcome in all cases of SIDS and near-miss SIDS may provide valuable insights into the optimal route for infant resuscitation. PMID- 11426481 TI - Chest compressions yielded higher airway pressures than Heimlich maneuvers when the airway was obstructed. PMID- 11426482 TI - On the existence of dragons. PMID- 11426483 TI - On the existence of dragons. PMID- 11426484 TI - Frank Pantridge and mobile coronary care. PMID- 11426485 TI - Pulmonary T cell repertoire in patients with graft-versus-host disease following blood and marrow transplantation. AB - Pulmonary inflammation is one of the risk factors associated with blood and marrow transplantation (BMT). To determine the potential role of T cells in pulmonary complications after transplantation, we analyzed the T-cell repertoire expressed in bronchoalveolar lavage fluids from eleven patients with graft-versus host disease following BMT. A reverse transcriptase-polymerase chain reaction was used to amplify rearranged TCR transcripts in unfractionated, CD4+, and CD8+ T cells from bronchoalveolar lavage fluids. The relative expression of TCR variable (V) gene families and the diversity of junctional region lengths associated with different AV and BV gene families were analyzed. Nearly all TCR AV and BV gene families were detected in bronchoalveolar lavage cells from BMT recipients. Oligoclonal patterns of TCR junctional region lengths were observed in unfractionated, CD4+, and CD8+ bronchoalveolar T cells. The oligoclonal expansion of bronchoalveolar T cells in patients was confirmed by DNA sequencing. TCRV gene expression is almost completely restored in the lungs of BMT recipients as early as two weeks after transplantation. Increased oligoclonality among TCR gene families suggests either an incomplete restoration of TCR diversity or an antigen driven expansion of T cells in the lungs of BMT recipients with graft-versus-host disease, not necessarily related to pulmonary infection. PMID- 11426486 TI - Role of transforming growth factor beta1 in microvascular endothelial cell apoptosis associated with thrombotic thrombocytopenic purpura and hemolytic uremic syndrome. AB - Primary human microvascular endothelial cells (MVEC) of restricted lineage undergo apoptosis when exposed to plasma from patients with thrombotic thrombocytopenic purpura (TTP) and sporadic hemolytic-uremic syndrome (HUS). This reflects the pathology and tissue distribution of lesions in vivo. As extracellular matrix (ECM) is critical to MVEC survival, and cytokines which regulate ECM, such as transforming growth factor (TGF)-beta1, have been reported anecdotally to be altered in TTP/HUS, we examined the role of TGF-beta1 and two ECM proteins, fibronectin and thrombospondin (TSP), in these disorders. Levels of active TGF-beta1 were elevated in acute but not convalescent phases of TTP/sporadic HUS, as well as TTP associated with human immunodeficiency virus infection and use of the anti-platelet drug ticlopidine. MVEC from tissues susceptible to TTP-mediated apoptosis showed little active TGF-beta1 production when exposed to TTP plasmas. In contrast, pulmonary MVEC and large-vessel EC, which are resistant to TTP-linked pathology, showed marked induction of TGF-beta1 following TTP plasma exposure. Exogenous TGF-beta1 suppressed TTP plasma-mediated apoptosis in susceptible MVEC in association with blockade of cell entry into S phase. Soluble TSP, devoid of detectable bound TGF-beta1, had a similar effect, which paralleled its ability to induce TGF-beta1 production in MVEC. In vivo, TSP deposition was diminished markedly in involved tissues of TTP patients. These data highlight the role of TGF-beta1 and ECM in TTP and suggest that differential production of TGF-beta1 by MVEC may play a role in their sensitivity or resistance to TTP/sporadic HUS-mediated apoptosis in vitro and in vivo. PMID- 11426487 TI - Phase II study of combination human recombinant GM-CSF with intermediate-dose cytarabine and mitoxantrone chemotherapy in patients with high-risk myelodysplastic syndromes (RAEB, RAEBT, and CMML): an Eastern Cooperative Oncology Group Study. AB - A Phase II study of GM-CSF with intermediate-dose cytarabine and mitoxantrone was conducted in patients with high-risk myelodysplastic syndrome. It was designed to evaluate if priming with growth factor could increase the efficiency of chemotherapy. In this older population only two of 10 patients achieved a bone marrow CR, including one patient whose leukemic blasts had an "S" phase increase of 2.55x at 48 hr. Unexpected hepatotoxicity was noted. This regimen cannot be recommended for this elderly population of patients. PMID- 11426488 TI - Primary thrombophilia in Mexico. II. Factor V G1691A (Leiden), prothrombin G20210A, and methylenetetrahydrofolate reductase C677T polymorphism in thrombophilic Mexican mestizos. AB - We have shown that in Mexican mestizo patients with clinical features of primary thrombophilia, 39% have activated protein C resistance phenotype, 5% protein C deficiency, and 2% protein S deficiency. In the present study, in a group of 37 thrombophilic Mexicans and 50 normal controls, we assessed the factor V G1691A (Leiden), the prothrombin G20210A, and the methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphisms. Four patients were found to be heterozygous for factor V Leiden, 5 heterozygous for the prothrombin 20210, 16 heterozygous, and 6 homozygous for the MTHFR 677. There were four individuals with co-segregation of alleles: two heterozygotes for the factor V Leiden/prothrombin 20210, one heterozygote for prothrombin 20210/MTHFR 677, and one heterozygote for prothrombin 20210/homozygote for MTHFR 677. For factor V Leiden, prothrombin 20210, and MTHFR 677 mutations, the allele frequencies were respectively 1% (+/ 0.2%, alpha = 0.05), <1% and 51% (+/-5%, alpha = 0.05), with calculated relative risks for thrombosis of 5.94 (P = 0.08), >7.66 (P < 0.05), and 0.44 (P NS), respectively. In Mexican mestizo thrombophilic patients, the low prevalence of the factor V Leiden mutation (10.8%) and the high prevalence of the prothrombin 20210 mutation (13.5%) contrast with those identified in Caucasian thrombophilic patients (21% and 6%, respectively; P < 0.01). On the other hand, the high prevalence of the MTHFR 677 mutation gene both in normal controls (78%) and thrombophilic patients (61%) does not support a role of this mutation in the thrombogenesis of Mexican mestizo patients. PMID- 11426489 TI - Prevalence of HTLV-I-associated T-cell lymphoma. AB - In order to assess the prevalence rate of HTLV-1-associated T-cell lymphomas and human retrovirus infection in general, approximately 21,000 individuals representing various patient populations, retroviral risk groups, and blood donors were examined for HTLV-I, HTLV-II, HIV-1, or HIV-2 infection using serologic and PCR assays. The prevalence rates among volunteer blood donors were 0.02% and 0% for HTLV and HIV, respectively. Significantly increased HTLV prevalence rates were observed among paid blood donors, African American health care clinic patients, Amerindians, recipients of HTLV-positive cellular blood products, intravenous drug users, sexual contacts and family members of HTLV positive people, and patients with primary thrombocytosis and other-than-low grade non-Hodgkin's lymphoma (NHL). Among some of these groups there were significant differences in the prevalence of HTLV-I versus HTLV-II. The eight HTLV-positive NHL patients all had mature, high-grade, CD4+ T-cell lymphomas with clonally integrated HTLV-I, for a prevalence of 4% among other-than-low-grade NHL patients. Seven of the eight died from their disease within 2 years despite treatment. Interestingly, two groups at risk for HTLV infection, namely needle stick victims and recipients of HTLV-infected and/or pooled plasma products, showed no evidence for infection. Significantly increased HIV-1 prevalence was observed among paid blood donors, African Americans, homosexuals, female prostitutes, hemophiliacs, and other-than-low-grade NHL patients. Only one patient was infected with HIV-2. Of the nine HIV-positive, other-than-low-grade NHL patients, seven HIV-1 positives had B-cell lymphomas, one HIV-1 positive had an HTLV-I-positive CD4+ T-cell lymphoma, and one infected with HIV-2 had a CD4+ T cell lymphoma that was HTLV negative. The data indicate that HTLV-I lymphoma, while uncommon, is not necessarily rare among other-than-low-grade NHL cases in the United States and, given its poor prognosis, should probably be studied separately in clinical trials. PMID- 11426490 TI - CD20-positive adult T-cell leukemia. AB - A 67-year-old woman was admitted to our hospital because of lymphadenopathy and lymphocytosis. Monoclonal integration of HTLV-I provirus DNA was detected, and a diagnosis of adult T-cell leukemia (ATL) was made. Flow cytometry revealed that the ATL cells expressed CD20 as well as T-cell-associated antigens, and expression of CD20 mRNA was also demonstrated. A novel T-cell subpopulation expressing CD20 molecules has recently been identified. This is the first report of CD20-positive ATL, suggesting that HTLV-I can infect and transform CD20 positive T cells. PMID- 11426491 TI - EBV-associated perianal Hodgkin's disease in an HIV-positive individual. AB - Hodgkin's disease (HD) has a higher incidence in HIV-positive individuals. It tends to occur at extranodal sites, frequently exhibits an unfavorable histological type with large numbers of neoplastic cells, and almost invariably harbors Epstein-Barr Virus (EBV). We describe a case of a 33-year-old HIV positive man who presented with anal pain from a 4-cm mass in the anorectal canal. He had no B symptoms or peripheral lymphadenopathy. A chest X-ray was within normal limits. A biopsy showed an ulcerated mass composed of a mixed infiltrate of lymphocytes, plasma cells, eosinophils, and Reed-Sternberg (RS) cells positive for CD15 and strongly positive for CD30. They were negative for CD45 and CD20. Numerous RS cells and lymphocytes were positive for EBV RNA using the EBER-1 probe. This highly unusual presentation of HD may reflect the greater incidence of anorectal lymphoma and of extranodal HD in the HIV-positive population. PMID- 11426492 TI - "Liver function tests" are not always tests of liver function. AB - A child with Wilm's tumor and a child with immune thrombocytopenic purpura (ITP) were each noted to have persistent elevations of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH). Both children underwent thorough evaluation for liver disease and, as a result, experienced delays in treatment of the Wilm's tumor and ITP. Eventually both children were found to have extremely elevated serum creatine kinase (CK). Muscle biopsy confirmed diagnoses of Duchenne's muscular dystrophy in one child, and Becker's muscular dystrophy in the second. Hematologists/oncologists should consider obtaining a serum CK to rule out muscle disease in patients with unexplained elevations of AST, ALT, and LDH. PMID- 11426493 TI - Anaplastic large-cell lymphoma which showed severe inflammatory status and myelodysplasia with increased VEGF and IL-6 serum levels after long-term immunosuppressive therapy. AB - We report a patient with anaplastic large-cell lymphoma (ALCL) who has been given immunosuppressive therapy for Evans syndrome for 10 years. He was admitted with spike fever, intra-abdominal lymphadenopathy, and multiple liver masses. Examination of biopsy specimens obtained by para-aortic lymph nodes and liver masses resulted in a diagnosis of ALCL. Immunohistochemically, these cells were reactive to anti-CD30 antibody but were not of B- or T-lineage. Bone marrow aspiration demonstrated the invasion of giant neoplastic cells and trilineage myelodysplasia. Because the patient showed severe inflammatory symptoms, we examined serum levels of various cytokines. Pretreatment levels of IL-6 and VEGF in this patient were significantly elevated compared to those of normal controls. He was treated with combination chemotherapy (ABVD regimen), achieving complete remission. Myelodysplasia and serum IL-6 and VEGF also normalized after treatment. We assumed that ALCL resulted from long-term immunosuppressive therapy and that the up-regulation of IL-6 and VEGF played a role in pathogenesis of this type of lymphoma. PMID- 11426494 TI - Possibility of potential VWD misdiagnosis or misclassification using LIA technology and due to presence of rheumatoid factor. AB - von Willebrand's disease (VWD) is now recognised to be the most common inherited bleeding disorder and is due to defects and/or deficiencies in von Willebrand factor (VWF). The latex immuno-assay (LIA) procedure has become a popular VWF:Ag detection methodology because of the ability to automate testing. In this report, we present findings which urge caution when normal LIA results are obtained co incident to striking clinical findings strongly suggestive of VWD, or previous laboratory findings consistent with VWD. As illustrated by a relevant case study, normal LIA results may lead to an "incorrect diagnosis" of "not VWD" or to a potential subtype misdiagnosis, should they be accepted without cross confirmation using alternative VWF methodologies. PMID- 11426495 TI - Behcet's disease in a patient with chronic myelogenous leukemia under hydroxyurea treatment: a case report and review of the literature. AB - Behcet's disease (BD) is a chronic, relapsing vasculitis of unknown etiology. Its association with chronic myelogenous leukemia (CML) is extremely rare, and typical manifestations of BD were observed in a very few patients with CML, mainly under interferon-alpha (IFN-alpha) treatment. Skin pathergy test, being positive in about 50% of patients with BD, is also positive in some IFN-alpha treated patients with CML without any evidence of BD symptoms. We describe a 62 year-old woman with CML who developed characteristic features of BD, including a positive skin hyperactivity test, during treatment with hydroxyurea. Hydroxyurea has been implicated in the appearance of skin vasculitic ulceration, but this is the first case, according to our knowledge, where the development of BD was observed during hydroxyurea maintenance in the chronic phase of CML. PMID- 11426496 TI - Plasma levels of vascular endothelial growth factor (VEGF) and its receptor, Flt 1, in haematological cancers: a comparison with breast cancer. AB - Raised plasma VEGF is found in some cancers but levels of its receptor soluble Flt-1 (sFlt-1) are unreported. Hypothesising increased levels to be present in haematological cancers, we measured both by ELISA in 22 patients with haematological cancer, 22 with breast cancer, and in age- and sex-matched controls. VEGF was raised in both patients groups compared to controls (P < 0.01) but was higher in haematological cancer compared to breast cancer (P = 0.0238). There was no difference in levels of sFlt-1. Our data point to changes in levels of plasma VEGF, but not sFlt-1, in haematological cancer that may have pathophysiological consequences. PMID- 11426497 TI - Satisfactory remission in a case of IgD myeloma: effectiveness of glucocorticoid treatment. AB - IgD myeloma is an infrequent type of multiple myeloma and is characterized by an aggressive clinical behavior and a short survival time. No satisfactory treatments have thus far been established. Recently we treated a patient with IgD(lambda)-type myeloma with glucocorticoids, and succeeded in achieving a complete remission. This case seems to indicate a usefulness of glucocorticoid monotherapy for treating IgD myeloma. PMID- 11426498 TI - T-gamma gene rearrangement and CMV mononucleosis. AB - A clonal T-gamma rearrangement was found in peripheral blood and bone marrow in a 57-year-old female who presented with 6-week history of fevers, night sweats, and weight loss. Splenomegaly, hemolytic anemia, atypical lymphocytosis, a marrow lymphoid aggregate, and elevated LDH had suggested lymphoproliferative disease. However, IgM serology for cytomegalovirus (CMV) was positive. With observation alone, her clinical features improved over 4 weeks with normalization of the blood count and disappearance of CMV viremia and the aberrant T-gamma clone. Acute CMV infection may mimic lymphoproliferative disease. T-gamma gene rearrangement may be part of the immune response to CMV infection and is not specific to lymphoid neoplasia. PMID- 11426499 TI - Mutations in the factor XI gene. PMID- 11426500 TI - Failure of recombinant FVIIa as treatment for abdominal bleeding in acquired hemophilia. PMID- 11426501 TI - Renal abnormalities in sickle-cell beta-thalassemia. PMID- 11426502 TI - Detection of pulmonary relapsed T-cell lymphoma by T-cell receptor (TCR) gene analysis. PMID- 11426503 TI - Splenic rupture as the presenting symptom of blastic crisis in a patient with Philadelphia-negative, bcr-abl-positive ET. PMID- 11426504 TI - High-dose cyclophosphamide in the treatment of severe aplastic anemia in children. PMID- 11426505 TI - Depression in later life: three etiologically different subgroups. AB - BACKGROUND: Various studies support the notion that early onset depression and late onset depression have different etiological pathways. Late onset depression has been found to be a heterogeneous group. This study attempts to divide the late onset group in two subgroups with different aetiology and find evidence for the vascular depression hypothesis. METHODS: Subjects were 132 depressed elderly persons from the general population, general practitioners and mental health care outpatient clinics. Sixty-four had early-onset depression (< 60), 69 had late onset depression (> or = 60). The latter group was divided into subjects with (n = 15) and without (n = 15) severe life stress. The groups were compared with respect to a variety of variables including vascular risk factors RESULTS: Early onset depression was associated with neuroticism and parental history. Subjects with late-onset depression and no severe life stress had higher vascular risk factors than whose depression was preceded by a severe stressor did. CONCLUSIONS: Our findings suggest that late life depression can be divided into subgroups with different etiological pathways: (1) early-onset with longstanding psychobiological vulnerability; (2) late-onset as reaction to severe life stress; and (3) late-onset with vascular risk factors. PMID- 11426506 TI - Sertraline versus imipramine to prevent relapse in chronic depression. AB - BACKGROUND: Chronic depressions are common, disabling and under-treated, and long term treatment is little studied. We report the continuation phase results from a long-term treatment study. METHODS: After 12 weeks of acute phase treatment in a double-blind, randomized, parallel-group, multi-center trial of sertraline or imipramine, patients with chronic depression (> or = 2 years in major depression, or major depression superimposed on dysthymia) continued study drug for 16 weeks. Initially, 635 patients were randomized to sertraline or imipramine in a 2:1 ratio. Nonresponders after 12 weeks entered a 12-week double-blind crossover trial of the alternate medication. Entry into continuation treatment required at least a satisfactory response (partial remission) to initial or crossover treatment. RESULTS: Of 239 acute or crossover responders to sertraline, 60% entered continuation in full remission and 40% with a partial remission. These proportions were identical for imipramine patients (n = 147). For both drug groups, over two-thirds of those entering in full remission retained it. For those entering in partial remission, over 40% achieved full remission. Patients requiring crossover treatment were less likely to maintain or improve their response during continuation treatment. The two drugs did not differ significantly in response distribution, drop out rates or discontinuation due to side effects during continuation treatment. LIMITATIONS: The absence of a placebo group constrains interpretation of our results, but chronic depressions have low placebo response rates. CONCLUSIONS: Most chronic depression patients who remit with 12 weeks of sertraline or imipramine treatment maintain remission during 16 weeks of continuation treatment. Most patients with a satisfactory therapeutic response (partial remission) after 12 weeks of treatment maintain it or further improve. Patients treated with imipramine experienced more side effects, but both drugs were well tolerated. PMID- 11426507 TI - Personality disorders in bipolar and depressive disorders. AB - The association of mood disorders with personality disorders (PDs) is relevant from a clinical, therapeutic and prognostic point of view. To examine this issue, we compared the prevalence of DSM-III-R personality disorders assessed with SCID II in patients with depressive (n = 117) and bipolar (n = 71) disorders both recovered from a major depressive index episode that needed hospital admission. PDs prevalence and comorbidity with axis I were calculated. Avoidant PD (31.6%) (O.R. = 1.7, C.I. = 1.06-2.9. P < 0.01), borderline PD (30.8%) and obsessive compulsive PD (30.8%) were the most prevalent axis II diagnoses among patients with depressive disorder. In bipolar disorder group, patients showed more frequently obsessive-compulsive PD (32.4%), followed by borderline PD (29.6%) and avoidant PD (19.7%). Avoidant PD showed a trend toward being significantly more prevalent among depressives (P < 0.07). A different pattern of PDs emerges between depressive and bipolar patients. PMID- 11426508 TI - Tryptophan hydroxylase gene 218A/C polymorphism is associated with somatic anxiety in major depressive disorder. AB - BACKGROUND: Abnormalities in functioning of the central serotonergic system have been implicated in the pathogenesis of depressive illness and suicidal behavior. Tryptophan hydroxylase (TPH) is the rate-limiting enzyme in the biosynthesis of serotonin, therefore, it may play an important role in regulation or control of serotonin functions. The aim of the present investigation was to determine whether there is an association between TPH gene polymorphism and major depression. particularly in patients with suicidal ideation. METHODS: A total of 135 unrelated patients suffering from major depressive disorder and 196 normal unrelated controls were included in the study. All controls and patients were Caucasian. A biallelic polymorphism at the tryptophan hydroxylase locus was genotyped. RESULTS: No significant difference between controls and depressed subjects in TPH gene polymorphism was detected. There was no association between TPH gene polymorphism and suicidal ideation. Total HAMD scores were not different between the genotypes or alleles in patients. However, among the HAMD clusters, somatic anxiety was significantly associated with TPH genotypes and alleles in that patients with 218A/A genotype had a significantly higher somatic anxiety scores compared to other genotypes. LIMITATION: Potential confounding effect of population stratification can not be excluded. The functional relevance of the TPH gene 218A/C polymorphism is, at present, uncertain. CONCLUSION: The polymorphism in serotonergic system related genes may be associated with depressive symptoms in major depressive disorder. The results suggest that analysis of clusters that narrow down the phenotype may be more suitable in genetic studies of major depressive illness. PMID- 11426509 TI - Depressive disorders and personality variables in HIV positive and negative intravenous drug-users. AB - BACKGROUND: Only a few reports investigated the prevalence of depression in intravenous drug-users with HIV infection, including both asymptomatic and symptomatic subjects. In the same group, the association of depression and personality diagnoses was also poorly researched. METHODS: A consecutive sample of intravenous drug-users was collected from patients admitted to an infectious disease clinic, another random sample was taken from out-patients attending a methadone maintenance treatment program. Subjects were first screened with the Hospital Anxiety and Depression Scale, and then all positive subjects were evaluated with the Composite International Diagnostic Interview. Depression was diagnosed according to DSM-IIIR. In-patients were also given a structured personality inventory (Karolinska Psychodynamic Profile). RESULTS: HIV-positive patients had a high rate of depression (major depression 36.2%, dysthymic disorder 7.1%) when compared to HIV-negatives (15.7 and 3.9%, respectively). In patients had the highest rate of depression, irrespective of HIV clinical staging. A personality disorder was diagnosed in 36% of the sample, but these subjects were no more significantly depressed. LIMITATIONS: Poor detection of depression by the admitting physician may have led to selective hospitalization of patients with both HIV and mood disorder. The composition of the sample may also be biased by the help-seeking behavior of HIV patients who are also depressed. CONCLUSION: Physicians treating AIDS patients should be alerted to the high rate of depression in clinical HIV illness, in order to identify and properly treat depression. PMID- 11426510 TI - Study of the influence of electroconvulsive therapy on the regional cerebral blood flow by HMPAO-SPECT. AB - STUDY OBJECTIVE: The aim of this preliminary study is to investigate the regional blood flow in response to ECT (electroconvulsive therapy) and to identify any responsive-pattern to the treatment. METHODS: STUDY DESIGN: Single longitudinal prospective study of cohorts. SUBJECTS: For this preliminary study ten patients, female sex, mean age 70.8 years with major mood disorder (CID-10 investigation criteria) were studied after signature consent. INTERVENTIONS: The intervention consisted in the administration of bilateral brief pulse ECT three times a week, during 6 to 12 sessions according to the standards of the Psychiatric Department of the Santiago Hospital in Victoria. MEASUREMENT: Clinical evaluation of depression was evaluated by Hamilton Depression Scale, Montgomery and Asberg Scale, Newcastle Scale and regional cerebral blood flow (rCBF) using the HMPAO SPECT. RESULTS: The pattern of distribution on the regional cerebral flow during the ECT showed changes from the basal pattern in all patients. All patients had a relative increased perfusion of the temporal lobes and basal ganglia. Other changes from the basal study were areas of decreased perfusion of the occipital lobe (6 patients) and parietal lobe (3 patients). CONCLUSIONS: Brain perfusion SPECT study of the patients with major depression shows changes during ECT. Further analysis are needed to understand the relationship between mechanisms of treatment and recovery in affective illness. PMID- 11426511 TI - Association between a catechol-o-methyltransferase polymorphism and obsessive compulsive disorder in the Afrikaner population. AB - BACKGROUND: It has been proposed that the catechol-o-methyl transferase gene (COMT) may play a role in the pathogenesis of obsessive-compulsive disorder (OCD). Whereas studies in a North American population showed that the low activity (L) allele of a functional polymorphism in COMT was associated with OCD in male patients, this result was not supported by studies in a Japanese population. The present association study assessed the risk for OCD conferred by this COMT polymorphism in a geographically different patient group, namely, the relatively genetically homogeneous Afrikaner population of South Africa. METHODS: Fifty-four unrelated OCD patients and fifty-four sex-matched controls were recruited from the same Afrikaner community. Patients and controls were phenotyped (DSM-IV) and genotyped for a NlaIII polymorphism with H (high activity) or L (low activity) alleles in the COMT gene. RESULTS: The H/L genotype was significantly more common than expected in the OCD patient group (P = 0.0017). LIMITATIONS: Replication studies with related individuals may be useful in discovering factors underpinning the H/L genotype abundance in the Afrikaner population. CONCLUSIONS: These results emphasise the need for further studies in genetically homogeneous populations to help define the complex etiology of this disease. PMID- 11426512 TI - The continuity of depression symptoms: use of cluster analysis for profile identification in patient and student samples. AB - BACKGROUND: Findings from several sources suggest that depression lies on a continuum whereby mild and severe variants are thought to differ in severity (i.e., quantitatively), but not in kind (i.e., qualitatively). The current study used cluster analysis to extend this work to examination of depression symptom profiles obtained in distressed student 'analogue' samples and clinically depressed samples. METHOD: Patients with major depressive disorder (n = 101) provided seed points for the depressed cluster, and 176 non-distressed university students (Beck Depression Inventory score < or = 8) provided seed points for the non-depressed cluster. The symptom profiles of three levels of analogue depressed samples were then evaluated (BDI > or = 9, BDI > or = 16, and BDI > or = 21). RESULTS: Only 35.4% of BDI > or = 9 analogue respondents were empirically sorted to the depression cluster and the majority were assigned to the non-depressed cluster. The proportion assigned to the depression cluster increased to 70.5% and to 86.2% when higher BDI cutoffs of 16 and 21 were examined, respectively. The DSM-IV depression symptom profile of the BDI > or = 21 group was very similar to the profile defined by clinical patients. LIMITATIONS: The study relied solely on self-report to assess symptom severity. CONCLUSIONS: It is recommended that higher BDI cutoffs be utilized in analogue depression research than is currently common. On quantitative grounds, analogue subjects who were sorted to the clinically defined depression cluster seem to best represent the idea of depression continuity. PMID- 11426513 TI - Atypical and non-atypical subtypes of depression: comparison of social functioning, symptoms, course of illness, co-morbidity and demographic features. AB - BACKGROUND: There are scant data regarding the demographic and psychosocial characteristics of outpatients with Atypical Depression (AD). METHODS: The demographic characteristics, rates of chronic dysphoria, baseline Symptom Check List Revised, and Social Adjustment Scale scores of 320 moderately depressed patients with and without AD were compared. RESULTS: ADs had a higher number of self-reported symptoms, greater impairments in functioning, and higher rates of chronic dysphoria and bipolar II than patients without Atypical Depression (NAD). LIMITATIONS: Variables used in this study were mostly cross-sectional, and the analyses were performed post-hoc. CONCLUSIONS: These data suggest ADs had a more pernicious course of illness than NADs, and that patients with AD were more symptomatic and dysfunctional at admission. PMID- 11426514 TI - Parent-child concordance for separation anxiety: a clinical study. AB - There is tentative evidence supporting a familial basis for separation anxiety. The present study aimed to examine parent-child concordance for that subtype of anxiety. Fifty-four children diagnosed with anxiety disorders and their parents (54 mothers and 29 fathers) were recruited from two juvenile anxiety clinics. Sixty-three percent of children diagnosed with juvenile separation anxiety disorder had at least one parent who suffered from the putative adult variant of the disorder (odds ratio = 11.1) (P < 0.001). Affected parents reported high levels of separation anxiety in their own childhoods. Juvenile separation anxiety disorder in children was not associated with any other parental diagnosis. The small sample size and other potential biases caution against definitive conclusions being drawn, but the present data add to existing evidence that separation anxiety may aggregate in families. PMID- 11426515 TI - A meta-analysis of the use of typical antipsychotic agents in bipolar disorder. AB - BACKGROUND: The potential benefits of typical antipsychotic agents in bipolar disorder are offset by serious treatment-associated side effects. Despite these concerns and the availability of mood stabilizing agents, the treatment of bipolar disorder with typical antipsychotic agents appears to be widespread. METHODS: A Medline search identified 16 publications that outlined medication use among 2378 bipolar disorder patients. Meta-analysis was used to estimate a weighted average of the relative proportions of the treatment use, where the weights were the reciprocals of the estimated variances for each study. RESULTS: Overall, 84.7% of bipolar patients received typical antipsychotic agents, with a loading toward a greater in-patient (90.7%) relative to out-patient (65.3%) use. Monotherapy accounted for 53.8% of typical antipsychotic use, and typical antipsychotic/mood stabilizer combination therapy accounted for 47.4%. In four studies where length of treatment data were available, the median of minimum typical antipsychotic use was 2.5 months, with 96.0% of the patients receiving typical antipsychotic agents. LIMITATIONS: The meta-analytic technique employed in this analysis is limited by the possible inclusion of studies with unreliable study designs or biased treatment practices, publication bias in which some studies may not have been reported, and possible lack of identification of all relevant studies. CONCLUSIONS: Typical antipsychotic agents are commonly used in the treatment of bipolar disorder, possibly due to dissatisfaction with mood stabilizer monotherapy especially in psychotic mania, the high prevalence of psychotic symptoms in acute mania, inappropriate continuation of typical antipsychotic agents after initial stabilization, and/or unavailability or unfamiliarity with new treatments. These findings also suggest that typical antipsychotics may have not only antipsychotic effects in mania but perhaps also antimanic properties. PMID- 11426516 TI - Antenatal depression, substance dependency and social support. AB - BACKGROUND: The purpose of this study was to explore the prevalence of depression and factors associated with depressive mood among pregnant women. METHOD: 391 women who were 14-37 weeks pregnant were evaluated with the Edinburgh Postnatal Depression Screen (EPDS), which has also been validated for prenatal use. Four questionnaires were used in order to explore associated factors: a questionnaire on background and pregnancy data, the Substance Abuse Subtle Screening Inventory (SASSI) and two Social Support Questionnaires (SSQ1 and 2). RESULTS: 7.7% of the total sample screened positive on the EPDS with a cut-off point of 12/13 recommended. Substance dependency and experienced difficulties in social environment had an independently significant association with maternal depression. LIMITATION: The caseness was defined with a self-report instrument. CONCLUSION: Substance dependency and experienced difficulties, especially in relation to friends, partner and own mother, are associated with antenatal depression. It is important to be aware of this when developing interventions in maternity care primary units. PMID- 11426517 TI - Effect of lithium and sodium valproate ions on resting membrane potentials in neurons: an hypothesis. AB - In an attempt to understand the therapeutic effects of lithium and sodium valproate in stabilizing the moods in manic depressive illness, the well-known Goldman-Hodgkin-Katz (G-H-K) equation is modified to include a fourth ion, such as a lithium ion or a sodium ion. The modified G-H-K equation is used to calculate the resting membrane potential in neurons. These calculations show that the resting membrane potential is depolarized depending upon the relative concentration of the lithium ion and upon its relative permeability. These calculations suggest that the resting membrane potential may be hyperpolarized in bipolar patients before treatment, and that the lithium ion perhaps depolarizes the resting membrane potential back to the normal level. They further support the prevailing hypothesis that manic-depressive illness may be caused by the hyperpolarization of the resting membrane potential, which, in turn, may be caused by the changes in ionic conductance (permeability) of the membranes. Sodium ions in sodium valproate do not significantly affect the resting membrane potential since they do not significantly change in the serum. PMID- 11426518 TI - Phenotypic characterization of subsets of T cell lymphoma: towards a functional classification of T cell lymphoma. AB - T cell non-Hodgkin's lymphomas (T-NHL) have traditionally been classified according to a variety of criteria including histological and clinical features, sites of involvement and etiologic agents. Except in select T-NHL types (e.g. CD30-positive anaplastic large cell lymphoma (ALCL)), immunophenotypic criteria are not used for routine subclassification of T-NHL. In this article. we outline the current models for classification and diagnosis of T cell tumors. We also briefly review the current understanding of non-neoplastic T cell subsets with regards to expression of activation markers belonging to the tumor necrosis factor receptor (TNFR) gene family. We summarize the currently available information on expression of these subset markers in T cell tumors, focusing on TNFR family members CD30 and CD134/OX40. CD134/OX40 expression is characteristic of certain entities (angioimmunoblastic lymphoma, angiocentric T-NHL) and a subset of T-NHLs of unspecified type, whereas CD30 expression is characteristic of ALCL and a largely non-overlapping subset of T-NHLs of unspecified type. Immunophenotypic stratification of T-NHL, using TNFR family members and other T cell subset-specific gene products, may provide a functional model for T-NHL classification as is currently the case for B cell tumors. PMID- 11426519 TI - Prognostic implications of p27 and cyclin E protein contents in malignant lymphomas. AB - The G1/S transition in the cell cycle is one of the checkpoints that can be deregulated in tumor development potentially causing increased proliferation and impaired capacity to arrest genetically damaged cells. The balance between activating and inhibitory molecules acting in the check point area seems to be critical and overexpression of cyclins and/or downregulation of the cdk inhibitors have been observed in many malignancies including lymphomas. In this review we have focused on two of the interplayers in the G1/S transition namely cyclin E and p27, and present the current knowledge of aberrations affecting these proteins in lymphomas as well as associations with clinico-pathological data including survival. PMID- 11426520 TI - The clinical significance of caspase regulation in acute leukemia. AB - Disruptions of pathways of programmed cell death, or apoptosis, are increasingly found in malignant cells of both solid and hematologic neoplasms. Caspases belong to a family of cysteine proteases and have emerged as central regulators of the apoptotic cascade. Despite many and diverse signals that can trigger apoptosis, the execution of apoptosis appears to be uniformly mediated through activation of caspase enzymes. Inapproriate expression of caspases or malfunctions in their regulation through other pathways may also be an important step in the pathogenesis of acute leukemias. Recent studies have shown that overexpression of the inactive forms of caspases CPP32 (caspase 3) and ICH-1 (caspase 2) is frequently observed in the blasts of patients with acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). Many other enzymes involved in apoptosis are expressed at high levels in patients with acute leukemia. Whether this signals the capacity of leukemic cells to rapid induction of apoptosis with fast reduction of the burden of disease and favorable clinical outcome, or accumulation of inactive substrates that cannot be activated by lack of cellular mechanisms to do so, requires further investigation. With the identification of many other regulators of apoptotic activity in the leukemic cells, new targets for future therapy may be identified and many new insights can be gained in understanding the biological behavior of response and resistance to therapy as well as control and relapse from minimal residual disease. PMID- 11426521 TI - TT virus in hematological disorders and bone marrow transplant recipients. AB - TT virus (TTV) was cloned as a possible causative agent for non A to C posttransfusion hepatitis. Determination of the entire sequence of the virus revealed that the virus is the first human circovirus. The nucleotide sequence of TTV has a wide range of diversity and at least sixteen genotypes have been discovered to date. The prevalence of TTV infection in the normal population differs among countries, but exceeds 10% in several countries. Most of TTV infections are not associated with hepatitis, although there is evidence of TTV induced hepatitis, especially caused by TTV of genotype I. To determine whether TTV is replicated in the liver is important in order to show that TTV is really a hepatitis virus, because results of a study in bone marrow transplant (BMT) recipients suggested that TTV might be replicated mainly in the hematopoietic cells. The prevalence of TTV infection in patients with hematological disorders who regularly require blood products was extremely high, but most of the infections did not cause liver injury, even in BMT recipients. PMID- 11426522 TI - Resveratrol--from the bottle to the bedside? AB - Resveratrol, a naturally occurring plant antibiotic has been the focus of a number of studies investigating its biological attributes, which include anti oxidant activity, anti-platelet aggregation effect, anti-atherogenic property, estrogen-like growth promoting effect, growth inhibiting activity, immunomodulation, and chemoprevention. More recently, since the first report on the apoptosis inducing activity of resveratrol in human cancer cells, the interest in this molecule as a potential chemotherapy agent has significantly intensified. Not only has its role as an anti-cancer agent been corroborated, but the precise mechanism(s) of the anti-cancer activity of resveratrol is/are being elucidated. Our group has been active in studying the cross talk between the caspase family of proteases and mitochondria, in drug-induced apoptosis. In this regard, we have shown that the cancer preventive activity of resveratrol could be attributed to its ability to trigger apoptosis in human leukemia and breast carcinoma cells. The cytotoxicity of resveratrol is restricted against these transformed cell types due to its ability to selectively upregulate CD95-CD95L interaction on the tumor cell surface, unlike normal peripheral blood cells. Despite the involvement of the CD95 signaling pathway, apoptosis induced by resveratrol is not accompanied by robust caspase 8 activation, but involves mitochondrial release of cytochrome C and downstream activation of caspases 9 and 3. We also extrapolate these in vitro findings in a murine model of carcinogensis, and demonstrate in vivo induction of apoptosis in mouse skin papillomas. These findings highlight the chemotherapeutic potential of this polyphenolic compound. PMID- 11426523 TI - Incidence of post transplant myelodysplasia/acute leukemia in non-Hodgkin's lymphoma patients compared with Hodgkin's disease patients undergoing autologous transplantation following cyclophosphamide, carmustine, and etoposide (CBV). AB - Secondary malignancies, particularly myelodysplasia (MDS), are serious events following high dose therapy with autologous stem cell support. We observed a higher frequency of secondary malignancies in patients with Hodgkin's disease (HD) than in patients with non-Hodgkin's lymphoma (NHL) undergoing high dose therapy with the same non-TBI conditioning regimen. Three hundred patients with Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL) were treated with cyclophosphamide, carmustine and etoposide and autologous stem cell support from 1986 through 1994. Median follow up of survivors is 3.9 years. Five-year survival is 51% for HD and 48% for NHL. Eleven patients developed second malignancies (9/150 treated for HD vs. 2/150 treated for NHL) a median of 2.4 years from transplantation and 5.2 years from initial diagnosis. Six patients had myelodysplasia or acute leukemia (MDS/AML) and 5 had lymphomas or solid tumors. Actuarial risk of MDS/AML at five years for patients transplanted for non Hodgkin's lymphoma is 3% (95% CI 0.6-9.6%). HD patients had significantly different pretreatment characteristics than patients with NHL. A Cox model showed that greater number of prior relapses and prior radiation therapy were significant risk factors for the development of MDS/AML. These data suggest that CBV is associated with a lower risk of secondary MDS/AML than TBI containing regimens and that much of the risk is associated with the pre-transplantation therapy. The use of autotransplantation early in the course of therapy for relapsed lymphoma might prevent some cases of MDS/AML. PMID- 11426524 TI - Comparison of amphotericin B lipid complex (ABLC) vs. ambisome in the treatment of suspected or documented fungal infections in patients with leukemia. AB - Fungal infections remain a major cause of treatment failure and death in acute leukemia. New liposomal preparations of amphotericin B are now available. While less toxic, their comparative efficacy and toxicity profiles are unknown. In this study the comparative efficacy and safety of ABLC vs. AmBisome was evaluated in seventy-five patients with leukemia who developed 82 episodes of suspected or documented mycosis, and were treated (1:1) with either ABLC (n=43) or AmBisome (n=39). Both drugs were dosed accordingly from 3 to 5 mg/kg/day. Using an intent to-treat analysis, the overall response to therapy was 27/43 (63%) for ABLC and 15/39 (39%) for AmBisome (p=0.03). Median dose and duration of treatment was 10 days at 3 mg/kg for ABLC and 15 days at 4 mg/kg for AmBisome. Acute, not dose limiting infusion side effects were seen in 70% vs. 36% (p=0.002), ABLC vs. AmBisome. Increase of bilirubin > 1.5 times from baseline was 38% vs. 59%, ABLC vs. AmBisome (p=0.05). ABLC and AmBisome were equally effective for the treatment of suspected or documented fungal infections. While, acute infusion-toxicity was greater with ABLC, infusion toxicity requiring discontinuation was similar for both drugs. AmBisome was better tolerated than ABLC but was associated with mild abnormalities in liver function tests at the end of therapy. PMID- 11426525 TI - Limited efficacy of intensified preparative regimens and autologous transplantation as salvage therapy in high grade non-Hodgkin's lymphoma. AB - Between 9/86 and 6/98, 22 patients with relapsed or refractory high grade lymphoma received intensified preparative therapy and underwent autologous transplantation at a single institution. Two intensified preparative regimens were used--cyclophosphamide, etoposide, total body irradiation (CY-VP-TBI) (N=17) and cyclophosphamide, BCNU, etoposide (CBV) (N=5). For all patients undergoing autologous transplantation, 5 year actuarial survival (S) and 5 year event free survival (EFS) were only 18% +/- 8%. Treatment related mortality was 14% overall but only 8% in patients receiving G-CSF or GM-CSF. Survival was significantly inferior to the survival observed in a concurrent series of patients with intermediate grade lymphoma, 34% +/- 6%, p < .05. Using high dose therapy in conjunction with autologous transplantation at the time of relapse may not be as valuable a strategy in high-grade lymphoma as in intermediate grade lymphoma although most studies combine the two disorders. Alternative strategies for the use of transplantation in high grade lymphoma, such as the use of transplantation as consolidation therapy, need to be investigated. PMID- 11426526 TI - Continuous infusion of vincristine-doxorubicin with bolus of dexamethasone(VAD) alternated with CHEP in the treatment of patients over 60 years old with aggressive non-Hodgkin's lymphoma. AB - This prospective study was undertaken to evaluate the efficacy and toxicity of combination chemotherapy with alternating cycles of vincristine, doxorubicin and dexamethasone (VAD) and cyclophophamide, doxorubicin, etoposide and prednisone (CHEP) in patients over 60 years old with previously untreated and advanced non Hodgkin's lymphoma (NHL) of intermediate- and high-grade malignancy. Eighty one consecutive, patients with NHL referred from April 1992 to October 1997 to GOELAMS centers were enrolled in this study and their outcome updated to June 1, 1999. Of 81 enrolled patients, 77 were eligible and assessable for response. The median age was 70 years (61 to 78), 85.7% were stage III or IV, 39% were of performance status > or = 2, 27.3% > or = 2 involved extra-nodal sites and 57.3% had higher LDH levels than normal. The immunophenotype was B in 87% and T in 13%. Fifty-one (66.2%) patients received the scheduled eight cycles of therapy and treatment was withdrawn in only 6 patients (7.8%) because of toxicity. Neutropenia grade 3-4 occurred in 11.1% after VAD courses vs 40.6% after CHEP courses. The mean cumulative dose of doxorubicin was 269 mg/m2 and the relative dose intensity was 84%. The overall response and complete response rates were 66.2% and 51.9% respectively, and after a median follow-up of 52 months the 3 year overall survival (OS) and event-free survival rates (EFS) were 43.5% and 33.0% respectively. In multivariate analysis, OS and EFS were statistically influenced by IPI (p = 3 x 10(-3); p < 1 x 10(-4)) and phenotype (p = 2 x 10(-3); p < 1 x 10(-4)). Our findings support the alternation of 4 courses of VAD and CHEP as it is well tolerated in patients over 60 years old with advanced intermediate- or high-grade NHL and provides response and survival rates comparable to 6 courses of CHOP. PMID- 11426527 TI - Second primary tumors and immune phenomena after fludarabine or 2-chloro-2' deoxyadenosine treatment. AB - The purine nucleoside analogs fludarabine and 2-chloro-2'-deoxyadenosine display substantial activity in the treatment of various chronic lymphoproliferative disorders. Their major toxicities are primarily immunosuppression and myelosuppression. The profound influence of these drugs on the immune system has raised questions as to the emergence of secondary neoplasms or auto-immune disorders after their use. Based on a literature review and on personal observations, this article reviews the potential clinical importance of these concerns. PMID- 11426528 TI - Cladribine in the treatment of chronic lymphocytic leukemia. AB - Cladribine (2-chlorodeoxyadenosine, 2-CdA) is a nucleoside analog with substituted halogen atom at position 2 in its purine ring that makes it resistant to deamination by adenosine deaminase (ADA). 2-CdA is the drug of choice in the treatment of hairy cell leukemia, but it is also highly active in other low grade lymphoid malignancies including chronic lymphocytic leukemia (CLL). The results of the studies presented so far have shown that 2-CdA gives similar complete response (CR) rate and overall response (OR) rate to fludarabine but the influence of both agents on survival times of the patients with CLL is still uncertain. CR rate induced with 2-CdA is significantly higher than in the patients treated with conventional chemotherapy. In refractory or relapsed patients 2-CdA induces 31 to 68% of overall responses including CR in 4 to 31%. In previously untreated patients overall remission rates of about 56-82% have been achieved with 2-CdA alone. When 2-CdA was used as primary therapy the CR rate was also significantly higher and ranged from 10% to 47%. Patients who received 2-CdA as their initial therapy and experienced a response lasting at least a year may be successfully treated subsequently with the same agent. A second response has been achieved in 35 to 100% patients treated with this agent for the second time. Despite the fact that 2-CdA gives higher CR and OR rates than conventional chemotherapy, it has not been established whether it has any influence on survival time. However, cross resistance between 2-CdA and FAMP in CLL patients is evident in the majority of studies. Bone marrow suppression with anemia neutropenia and thrombocytopenia are the dose limiting factors for 2-CdA use. These side effects are pronounced in heavily pretreated patients and after multiple courses of therapy. Treatment with this agent also leads to the decrease of the CD4+/CD8+ ratio for an extensive period of time exceeding 12, even up to 24 months. In consequence, infections including opportunistic type, are frequently observed. We suggest, that in patients with CLL, 2-CdA should be used as second line treatment rather than the first line therapy until the final results of ongoing randomized clinical trials are available. PMID- 11426529 TI - Cost-Effectiveness of interferon alfa-2b added to chemotherapy for high-tumor burden follicular non-Hodgkin's lymphoma. AB - Recent data from GELF (Groupe d'Etude des Lymphomes Folliculaires) have shown that the addition of interferon alfa-2b (IFN) to a doxorubicin-containing regimen (CHVP: cyclophosphamide, doxorubicin, teniposide and prednisone) prolongs both progression-free survival and overall survival in high-tumor-burden follicular non-Hodgkin's lymphoma. This gain must be weighed against the incremental toxicity and cost of IFN over CHVP alone and the objective here was, to determine the marginal cost-effectiveness of additive IFN in the specific setting of high tumor-burden follicular non-Hodgkin's lymphoma. Meta-analysis of GELF trial results employing a Markov model was used with three health states: No Progression, Progressive Disease, and Death. Treatment response, survival and toxicity data are drawn from the GELF study. The current study is based on the final analysis of 242 patients (J Clin Oncol 1998;16:2332-2338), with a six year median follow-up for overall survival (median overall survival: not reached for CHVP + IFN vs 5.6 years for CHVP Only, p = 0.008). MEASUREMENTS: Quality of life data (utilities) are taken from studies with similar dosing of IFN, from Q-TwiST (quality adjusted time without symptoms or toxicity) analysis of the GELF data and from a panel of experts gathered to develop treatment models for high-tumor burden follicular non-Hodgkin's lymphoma. Costs and quality-adjusted years of life saved were discounted at 3% per annum. SETTING: Costs determined for university medical centers in the United States. Results showed that, at the median cohort age of 52, IFN add 9.9 quality-adjusted months at an added cost of $13,900 (marginal cost-effectiveness of $16,900 per quality-adjusted life year, or QALY). A more complex, two-stage model approximates the actual cohort survival curves much better than a simple, one-stage model, but both models yield essentially the same marginal cost-effectiveness. Sensitivity analysis to quality of life on IFN shows marginal cost-effectiveness ranging from $15,200/QALY (no penalty for IFN) to $21,300/QALY (20% quality adjustment, greater than that reported). The model is quite insensitive to the probability of IFN toxicity. The model is moderately sensitive to the efficacy of IFN in delaying progression, particularly in the first 18 months (pProgI), but the marginal cost-effectiveness does not rise to $50,000/QALY until pProgI increases 220% from the baseline. Although the model is moderately sensitive to the cost of IFN (cIFN), marginal cost-effectiveness is below $50,000/QALY for values of cIFN below $2580/month (baseline cIFN = $850/month, corresponding to a marginal cost-effectiveness of $16,900/QALY in the baseline case). If the model is modified to reflect the 14% overall survival advantage at five years found in trials utilizing more intensive initial chemotherapy (including the GELF trial), then the marginal cost effectiveness drops to $11,900/QALY in the baseline case. In condusion, based on data from the GELF study, low-dose interferon alfa-2b is cost-effective when added to CHVP therapy in the treatment of high-tumor-burden follicular non Hodgkin's lymphoma. The analysis is robust: the model employs very conservative assumptions, and additive IFN remains cost-effective over wide ranges of variables in sensitivity analyses. The marginal cost-effectiveness is best expressed as being in the range of $12,000/QALY to $17,000/QALY in the baseline case. A simple Markov model can be used to describe treatment regimens with distinct periods of therapy. PMID- 11426530 TI - Secondary chromosome changes in mantle cell lymphoma: cytogenetic and fluorescence in situ hybridization studies. AB - To better define the incidence and nature of secondary chromosome anomalies in mantle cell lymphoma (MCL) carrying the t(11:14)/BCL1 rearrangement, cytogenetic and fluorescence in situ hybridization studies (FISH) were performed in 42 patients (39 classical histology, 3 blastoid variant), using 6q21, 9p21/p16, 13q14, 17p13/p53 and chromosome-12-specific probes. Karyotypes from 89 cases published in 5 recent series including patients diagnosed in a homogeneous fashion were reviewed. In our series, FISH confirmed the interpretation of the karyotype in all cases and disclosed cryptic chromosome deletions in a sizeable fraction of cases. One patient (2.4% of total) was found with a cryptic 9p21 deletion by FISH. Two cases (4.8%) had a 6q21 deletion at CCA and at FISH; +12 was found in three cases by CCA plus nine by FISH (28.6%); 13q14 deletion was found in six cases by CCA plus 16 by FISH (52.4%), 17p13 deletion in three cases by CCA plus 8 by FISH (26.2%). In 131 patients (42 present series plus 89 in the literature) secondary chromosome aberrations seen by conventional cytogenetic analysis in more than 5 cases included deletions/translocations (del/t) 6q15-23 [15 cases]; -13 [14 cases]; del/t 1p21-31 [12 cases]; +3q [11 cases]; del/t 17p [9 cases]; 8p translocations and del(Y) [8 cases each]; -20 [7 cases]; 13q14 deletion, del/t 11q22-23, del/t 9q, del(10)(q22q24), -20, -21, -22 and -X [6 cases each]. We arrived at the following conclusions: i) though no secondary anomaly is specific for MCL, there is a distinct profile of recurrent chromosome lesions in MCL with 1p21-31 deletions, 8p translocations, 11q22-23 anomalies having a strong association with CD5+ B-cell lymphomas of low-to-intermediate grade histology; ii) FISH enabled the detection of cryptic chromosome 12, 13q and 17p rearrangements in a sizeable fraction of cases; iii) 9p21/p16 deletions did not occur at a high incidence in this series, possibly because of the low number of cases with blastoid variant. PMID- 11426531 TI - Seroprevalence of Helicobacter pylori infection in patients with lymphoma. AB - To determine the Helicobacter pylori (HP) seroprevalence in patients with non Hodgkin's lymphoma (NHL) and other hematological conditions. Sera were collected from 444 patients with NHL, Hodgkin's disease (HD), lymphoproliferative disorders (LPD), myeloproliferative disorders (MPD), and other hematological conditions. HP seropositivity was determined by ELISA and the results were compared among diagnostic groups HP seropositivity was observed in 168/444 (38%) of the total population. Higher seropositivity rates were associated with increasing age (p=0.001), and country of birth outside the USA and Canada (p=0.0001). Among the diagnostic groups, patients with NHL demonstrated the highest frequency (43%) and those with HD, the lowest frequency (20%; p=.026) of HP seropositivity. The differences among diagnostic groups remained statistically significant after controlling for country of birth (p<0.05), but not after controlling for patient age at diagnosis. The HP seroprevalence of G1 NHL was 55% compared to 40% for non G1 NHL (p=NS). The highest rate of HP seropositivity (67%) occurred in gastric MALT lymphoma patients, although this did not reach statistical significance compared to the non MALT group (50%) due to small sample size. In conclusion, the rate of HP seropositivity in patients with MALT lymphoma in the USA appears to be lower than in Europe. Helicobacter pylori does not appear to be an important factor in other types of NHL of the G1 tract or elsewhere. Studies of HP prevalence should be controlled for country of birth as well as for age. PMID- 11426532 TI - Establishment of a cytokine-producing anaplastic large-cell lymphoma cell line containing the t(2;5) translocation: potential role of cytokines in clinical manifestations. AB - A permanent cell line, HSC-M1, was established from a child with advanced CD30 (Ki-1)+ anaplastic large-cell lymphoma (ALCL). Clinical features included irritability, fever, weight loss, tender lymphadenopathy, pneumonitis, neutrophilia, and bone marrow erythrophagocytosis. While HSC-M1 cells exhibited an immunophenotype characteristic of ALCL of T-cell lineage, the cell line also demonstrated features of monocyte-macrophage lineage. Cytogenetic and polymerase chain reaction (PCR) analysis of the HSC-M1 cell line and involved bone marrow demonstrated the characteristic non-random chromosomal translocation t(2:5)(p23:q35). Reverse transcriptase PCR for mRNA expression of cytokines and cytokine receptors showed that HSC-M1 cells expressed the message for multiple cytokines and their receptors. Measurement of cytokine levels in serum samples using enzyme-linked immunosorbent assays showed increased concentrations of several cytokines. The increased levels of some cytokines correlated with disease activity and clinical symptoms. Although spontaneous production by HSC-M1 cells of some of these cytokines was demonstrated, the production of others was only detectable after stimulation with exogenous CD30 ligand. With few exceptions, there was good correlation between serum cytokine levels and cytokines produced by HSC-M1 cells. These findings indicate that cytokine production is a feature of ALCL cells and that some of the clinical manifestations in ALCL may result from cytokines produced by either the malignant or accessory cells. PMID- 11426533 TI - MDR-1 expression and deletions of chromosomes 7 and 5(Q) separately indicate adverse prognosis in AML. AB - In order to assess any correlation between MDR-1 expression and chromosomal aberrations, and to define their impact on clinical outcome in newly diagnosed AML pts, we investigated bone marrow and peripheral blood samples of 49 consecutive pts admitted to our hospital. Monosomy 7, trisomy 8 and 5q- were evaluated by means of interphase fluorescence in situ hybridization (FISH). Monosomy 7 was present in 6 pts, trisomy 8 in 5 pts, and 5q- in 6 pts. More than one aberration was seen in 7 pts. Chromosomal aberrations were mostly found in older pts (12/14 >60 years; p=0.03) and in pts with CD34 positive leukemic blasts (13/14 coexpressed CD34, p=0.0004). In 25 pts also standard G-banding analysis was performed leading to concordant results regarding chromosomes 7, 8 and 5. Flow cytometry identifyed MDR-1 positivity (MDR+) in 16 pts. MDR-1 expression appeared to be a characteristic feature in CD34+ AML (12/16 were CD34+ and MDR+ pts; p=0.013). No correlation, however, was found between chromosomal aberrations and MDR-1 expression. Pts with aberrations of either chromosomes 7, 8 or 5 detected by FISH (FISH+) were predominantly resistant to induction therapy (6/8 pts, p=0.004). A lower rate of complete remission (CR) was also seen in pts with MDR-1 expression (p=0.006). MDR+/FISH+ pts (n=3) were all refractory to remission induction, while all MDR-/FISH- pts (n=19) achieved CR (p=0.0006). MDR-1+ as well as pts with aberrations of chromosomes 7, and 5(q) showed a significantly decreased probability of overall survival. In conclusion, MDR-1 expression as well as abnormalities of chromosomes 7, and 5(q) predict poor clinical outcome in AML. The identification of these prognostic factors provides useful information for risk adapted treatment strategies. PMID- 11426534 TI - Adhesion molecule expression, clinical features and therapy outcome in childhood acute lymphoblastic leukemia. AB - In view of the relevance of adhesion molecule expression for the mechanisms of homing, trafficking and spreading of malignant cells, we have investigated the expression of surface adhesion molecules in lymphoblasts from 57 acute lymphoblastic leukemia (ALL) cases and tried to correlate the adhesive phenotype with immunological typing, prognostic factors at diagnosis and clinical follow up. Blasts from all cases expressed adhesion molecules at high rates. Beta1 integrin chain (CD18) was consistently found on blasts from most ALL cases: among integrins of the beta2 family. LFA-1 was detected in 58% of cases, in the virtual absence of other alpha chains. CD54 and CD58 were expressed in variable proportions by ALL blasts and CD44 was detected in the majority of the malignant cells, whereas the CD62L selectin was only present in 24% of cases. B-lineage ALL's displayed similar adhesion molecule phenotypes irrespective of maturational stages of the leukemic cells. We found a significantly reduced expression of beta2 alphaL integrins in the hybrid ALL cases (CD13 and/or CD33 positive). However, these cases did not show differences in clinical presentation and behaviour in comparison with patients of other groups. We did not find a significant correlation between adhesion molecule expression and well established risk factors (age, white blood cell count, central nervous system involvement, chromosomal abnormalities), with the exception of splenomegaly, that was significantly associated with CD18 expression. In the follow-up, no evidence of significant correlation between adhesive phenotype and adverse events such as leukemic relapse and death was found. In conclusion, although expression of adhesion molecules on lymphoblasts confirms the phenotypic heterogeneity of ALL, it appears that this is not relevant for the clinical aspects of the disease and for prognosis. PMID- 11426535 TI - VLA-4 mediated adhesion to bone marrow stromal cells confers chemoresistance to adherent lymphoma cells. AB - The mechanisms of maintenance of residual lymphoma in bone marrow during chemotherapy are currently not well understood. Previous studies have shown that primary lymphoma cells obtained from histologically negative bone marrow of non Hodgkin's lymphoma (NHL) patients grew in long-term bone marrow cultures primarily in association with bone marrow stromal cells. Furthermore, the interaction of NHL patient cells with bone marrow stromal cells inhibited their spontaneous apoptosis. The current studies were designed to characterize the components of the heterotypic interaction between lymphoma cells and bone marrow stromal cells as well as to probe the consequences of this interaction as it pertains to the potential survival of minimal numbers of lymphoma cells during chemotherapy. Cellular adhesion assays performed in the presence of either neutralizing antibodies to VCAM- or the alpha and beta subunit of VLA-4 resulted in >95%, 82% and 35% inhibition of lymphoma cell line adhesion to the bone marrow stromal line MS-5, respectively. Modulation of VLA-4 affinity by the 8A2 antibody resulted in enhanced secondary adhesion at 24 and 72 hours to either cellular fibronectin (65% and 65%) or MS-5 cells (60% and 55%), superceding levels obtained using untreated lymphoma cells (<20%). The bone marrow stromal cells induced a chemoprotective effect for adherent lymphoma cells over a 3-log dose range of vincristine, resulting in a 2-log increase in the ED50 at day 6 of culture. The failure of glutaraldehyde fixed stromal cells to induce a chemoprotective effect demonstrated that viable bone marrow stromal cells were necessary. Similarly, lymphoma/stromal cell conditioned medium also failed to provide a survival advantage. These data demonstrated that viable bone marrow stromal cells possessed the ability to actively inhibit the apoptotic pathways of intimately adherent lymphoma cells and this potentially contributes to their survival during chemotherapy. PMID- 11426536 TI - Retinoblastoma gene product and P21 (WAF1, CIP1) protein expression in non Hodgkin's lymphomas: a multivariate survival analysis. AB - We evaluated immunohistochemically the expression of two negative regulators of the cell cycle, namely retinoblastoma gene product (pRb) and WAF1/Cip1 gene product (p21), in paraffin sections from 93 patients with non-Hodgkin's lymphomas (NHL) and related it to clinicopathological parameters, proliferative fraction, p53 expression and survival. Patients were followed until death (n=33) or for an average of 52 months (60-160). Rb labelling index (LI) increased with malignancy grade and proliferative activity but was unrelated to other clinicopathological parameters. In 33% of cases, especially those of the aggressive groups, we observed diminished pRb expression (i.e. low pRb/Ki-67 ratio). p21 expression on the other hand correlated only with histological grade, Rb LI and p53 LI. In multivariate analysis, Rb LI was a negative predictor of disease-free survival but was linked to a higher probability of complete response. However, diminished pRb expression as well as p21 expression were not statistically significant prognostic indicators. Our results suggest that pRb as a cell cycle related molecule may play an important role in determining prognosis and therapeutic response in NHL patients. PMID- 11426537 TI - Lovastatin induced control of blast cell growth in an elderly patient with acute myeloblastic leukemia. AB - We recently reported that AML cells derived either from cell lines or from patients undergo apoptosis in response to lovastatin, an agent used extensively in the treatment of hypercholesterolemia. The concentration of lovastatin required to achieve this in culture varies from patient to patient, however, the in vitro concentrations required to kill AML cells, can be attained clinically. While in vitro studies assessing responsiveness of leukemic cells to lovastatin were being performed, a 72 year old female presented with relapsed AML. The patient did not desire any further induction therapy. As the patient's cells proved to be sensitive in culture to lovastatin, the patient was offered this drug. In this brief report we describe a case in which there was apparent control of the patient's leukemic blast cells by lovastatin at a dose double the usual recommended dose for hypercholesterolemia. This case illustrates the potential for lovastatin to provide a novel means of controlling leukemic cell growth in AML patients. PMID- 11426538 TI - Primary lymphoma of spermatic cord. AB - Primary lymphomas of spermatic cord are extremely rare. In a review of the world medical literature, until now, only fourteen cases of spermatic cord lymphoma have been reported, and, furthermore, they have a poor prognosis even in patients with stage I disease. Herein, we report a new case of primary non-Hodgkin's lymphoma of the spermatic cord. In August, 1993, 76-year-old man visited an urological hospital with a compaint of a right intrascoral mass, and underwent orchiectomy. Macroscopically no invasive lesion in the testis was observed, and the tumorous lesion was restricted to the epididymis. The histopathological study indicated that he suffered from primary malignant lymphoma of the spermatic cord (B-cell, diffuse medium-sized cell type). As radiographic investigations showed no other invasive lesion, the patient was diagnosed to be in stage IE. He was followed only with clinical observation, and, in August, 1996, relapsed with extensive disease in the abdoninal cavity, and was transferred to our hospital. Fourty months after the orchiectomy, he died of progression of disease irrespective of the salvage radio-chemotherapies given to him. PMID- 11426539 TI - Primary hepatic lymphoma. AB - We report the histologic and CT imaging findings of a patient with primary Burkitt's lymphoma of the liver. She presented with a massive, painless abdominal mass. The tumor responded to chemotherapy but the patient died of neutropenic sepsis. PMID- 11426540 TI - Molecular remission following high-dose hydroxyurea and fludarabine plus cytarabine in a patient with simultaneous acute myeloid leukemia and low-grade lymphoma. AB - The occurrence of acute myeloid leukemia (AML) as a secondary tumor has been frequently reported in patients who received various chemotherapy regimens for hematologic malignancies wile the concomitant development of chronic lymphoproliferative diseases (CLD) and AML in previously untreated patients is extremely rare. We report a case with an apparently spontaneous occurrence of AML and non Hodgkin low-grade lymphoma diagnosed by immunological, cytogenetical and molecular analyses. In particular genetic studies allowed to identify the coexistence of a clonal lymphoid population and a myeloid blast component characterized by inv(16) marker and CBFbeta-MYH11 gene fusion. Complete remission of AML and the CLD was obtained following high doses of hydroxyurea and two consolidation cycles of fludarabine plus intermediate dose cytarabine. PMID- 11426541 TI - Mycobacterial pulmonary infection post allogeneic bone marrow transplantation. AB - Allogeneic bone marrow transplant recipients are prone to pulmonary infections caused by a wide spectrum of organisms. Since the first bone marrow transplatation (BMT) done in 1983 at the Tata Memorial Hospital, we have recently seen the first case of Mycobacterium Fortuitum Chelonae complex among 117 BMT (including 90 allogeneic and 27 autologous) patients. The patient was on immunosuppressants for chronic GVHD post allogeneic BMT done for CML-CP. He developed pulmonary mycobacterial infection 13 months post BMT. Diagnosis was difficult because of the atypical presentation, negative culture reports, and the presence of multiple pathogens due to immunosuppression. In our case the diagnosis was eventually established after examination of material obtained by bronchoscopy. Patient has shown response to antituberculosis drugs after 2 months. This shows the need to consider atypical mycobacterial infection in the differential diagnosis of pulmonary illness in the post allogeneic BMT setting. PMID- 11426542 TI - Central nervous system relapse in a patient with mantle cell lymphoma in continuous clinical and molecular remission at six years since autografting. AB - Although molecular remissions have been frequently observed and associated with low likelihood of relapse in some lymphoid tumours, they are seldom reported in mantle cell lymphoma (MCL). We performed PCR analysis of a MCL patient with central nervous system (CNS) relapse occurring 76 months after autologous transplantation. Molecular follow-up showed constant absence of PCR-detectable disease, even after the onset of relapse. These data indicate that isolated CNS relapse may occur even after several years of continuous remission and cannot be excluded based on a persistent pattern of molecular remission. However, the prolonged remission duration observed in this patient suggests that achieving PCR negativity may also be of benefit for MCL patients. PMID- 11426543 TI - Teaching cases from the Royal Marsden and St Mary's Hospitals. Case 20. Renal impairment in non-Hodgkin's lymphoma with C1 inhibitor deficiency. PMID- 11426544 TI - Hematopoietic stem cells can be CD34+ or CD34-. AB - Until recently, it was thought that the most primitive HSC have a fixed phenotype within a hierarchical differentiation system, and that changes in engraftment and renewal potential occur in a stepwise fashion linked with differentiation. In this review, we summarize the data from several different species and different animal models of hematopoietic stem cell function. Taking into account all of the published data, it becomes clear that the hematopoietic stem cell compartment contains more than one phenotypically identifiable population capable of self renewal and long term pluripotent engraftment. It is clear that some stem cells express CD34, and others do not. The exact phenotypic progression between these cells needs to be further defined, because different in vivo and ex vivo manipulations may shift the stem cells from one phenotype to another, and this can complicate interpretation of experimental transplant data. PMID- 11426545 TI - Deletions within the epstein-barr virus latent membrane protein-1 oncogene in adult ordinary, HIV-associated and paediatric Hodgkin's disease. AB - The aims of this study were the following: a) to perform Epstein-Barr virus (EBV) strain type assignment in three groups of Hodgkin's disease(HD): adult ordinary (39 cases), paediatric (24 cases), and HIV-associated (30 cases) and to compare the prevalence of type 1 and type 2 in each of the groups with that existing in two reference populations made up of 50 adults and 39 children; b) to assess the frequency of latent membrane protein-1 (LMP-1) 30-base pair (bp) deletions in the HD groups and in the healthy controls; and c) to relate the presence of LMP-1 deletions with EBV type. Type 2 EBV was observed in 12.8% of ordinary HD, in 26.7% of HIV-associated HD, in 25% of paediatric HD, in 4% of adult controls, and in none of the healthy children. The existence of double infections by type 1 and 2 EBV was also observed in 5.1% of ordinary HD, in 6.7% of HIV-associated HD, and in 10% of adult controls. The 30-bp deletion was identified overall in 33.3% of ordinary HD, in 83.3% of HIV-positive HD, 79.2% of paediatric HD, 34.7% of adult controls, and 36.4% of healthy children. Statistical analysis showed a significant association of the deleted strains with HD occurring in HIV-positive patients (P= 0.00003) and childhood HD (P= 0.006). On the other hand, the prevalence of the 30-bp deletion in the adult ordinary HD group reflects the prevalence of the deletion in the general population. Co-infections by deleted and non-deleted EBV strains were detected in 12.8% of ordinary HD, in 33.3% of HIV-associated HD, in 50% of paediatric HD, in 26.5% of adult controls, and in 27.3% of healthy children. Concerning the relationship between the deletion and the EBV typing, 26% of type 1 specimens carried the 30-bp deletion in an isolated manner compared with 64.7% of type 2. The statistical analysis showed that the deletion was associated with type 2 strains when coinfections were excluded and only the cases in which the deletion appeared alone were considered (P=0.003). PMID- 11426546 TI - Contribution of nitric oxide to the apoptotic process in human B cell chronic lymphocytic leukaemia. AB - B cell chronic lymphocytic leukaemia (B-CLL) is characterised by defective apoptosis that cannot be explained solely on the basis of the known chromosomal abnormalities. We and other have now reported that the leukemic cells spontaneously display the inducible isoform of nitric oxide synthase, iNOS. Inhibition of the iNOS pathway leads to increased apoptosis of the tumoral cells in vitro, indicating that the endogenous release of NO contributes to their resistance to the normal apoptotic process. The factors that induce the expression of iNOS in vivo in the leukemic cells are not yet identified. Yet, as interaction of B-CLL leukemic cells with bone marrow stromal cells promotes their survival, the involvement of adhesion molecules and integrins may be suspected. The engagement of CD23 stimulates iNOS activation in the tumoral cells, suggesting that in vivo interaction of CD23 with one of its recognised ligands may contribute to iNOS induction. A role for CD40-CD40 ligand interaction may also be hypothesised. The mechanisms involved in the anti-apoptotic role of NO are not fully understood, but may implicate the inhibition of caspase activity, hence the impairment of the Fas pathway. In addition, the mitochondrial membrane potential disruption appears to be a NO-sensitive step in the apoptosis cascade. The presence of a NOS displaying anti-apoptotic properties has now been recognised in different cell types, including various leukaemia. A better knowledge of the mechanisms governing the ultimate fate of NO, anti- versus pro apoptotic would allow the development of new therapeutic approaches for the treatment of these diseases. PMID- 11426547 TI - 11q deletions in hematological malignancies. AB - Structural aberrations involving 11q are among the most common aberrations in a number of hematological malignancies. Most of the aberrations, such as translocations and deletions, often harbor a breakpoint at 11q23, which suggests that this region might contain a tumor suppressor gene important for the genesis of lymphoproliferative disorders. Interestingly, deletions are concentrated only in some subtypes of hematological malignancies, where they are detected at a relatively high frequency. In B-cell chronic lymphocytic leukemia (B-CLL), deletions have been detected in 20-30% of the cases, whereas almost half of the mantle cell lymphomas (MCL) show deletion at 11q23 in fluorescence in situ hybridization analysis. In T-cell prolymphocytic leukemia (T-PLL), deletions involving the region 11q23.3-23.1 have also been detected to be frequent. In B cell chronic lymphocytic leukemia, 11q deletion is associated with more rapid disease progression and poor survival in a younger subgroup of patients. The putative tumor suppressor genes have remained unrevealed until recently, when the ATM gene was found to carry mutations in cases with deletion in B-CLL, MCL and T PLL. These data suggest that 11q deletions and dysfunction of the ATM gene might have significance in the tumorigenesis of certain subsets of hematological malignancies. Importance of 11q deletion as a diagnostic marker needs to be further studied in a larger series of patients. Another issue that remains to be investigated is the involvement of other target genes in the deletion. PMID- 11426548 TI - HTLV-I Tax related dysfunction of cell cycle regulators and oncogenesis of adult T cell leukemia. AB - HTLV-I is causually related to the oncogenesis of adult T cell leukemia (ATL). However, the precise mechanism of HTLV-I oncogenesis is unclear. HTLV-I Tax protein functions as an activator of various cellular genes, including IL-2, IL-2 receptor-alpha, and c-fos through the activation of nuclear transfer factors such as NF-kappaB and SRF, and also potently activates trascription of viral genes through CREB/ATF sites in the viral LTR. However, Tax activation of HTLV-I infected T cells through the above pathways induces polyclonal proliferation of the cells in vitro; Tax however may function only transiently in the immediate post-infection period following infection in vivo. The long latent period of 60 years from infection to onset of disease suggests other mechanisms for ATL oncogenesis. Recent studies suggest that the malignant transformation of ATL is a multi-hit phenomena, suggesting that discrete genetic events are responsible for ATL oncogenesis. These genetic events could be responsible for the different stages of ATL: smoldering, chronic, lymphoma, and acute type, p16 and p53 genes are important negative regulators of the cell cycle and are often found to be mutated in neoplasms. Recent studies including ours demonstrated a high frequency of alteration of these two genes in primary ATL cells. Furthermore, alteration of the two genes is associated with acute but not chronic type ATL. In addition, p16 gene alteration is linked to the growth rate of ATL cells, suggesting that the alteration of these cell cycle regulatory genes may be related to progression from smoldering or chronic to acute or lymphoma type ATL. Tax may be involved in mutagenesis of these genes through suppression of DNA-beta polymerase gene expression during the process from latent period to acute/lymphoma type. Once transformation occurs, activation of the pathway between Tax and the three nuclear transfer factors, NF-kappaB, SRF, and CREB/ATF, contributes to establish the aggressive manifestations of acute/lymphoma type ATL cells. PMID- 11426549 TI - Bone marrow leukemic progenitor cell content in pediatric T-lineage acute lymphoblastic leukemia patients with an isolated extramedullary first relapse. AB - Isolated extramedullary relapse in childhood acute lymphoblastic leukemia (ALL) is associated frequently with the T-lineage immunophenotype and may be accompanied by occult bone marrow disease. We employed highly sensitive multiparameter flow cytometry and blast colony assays to quantify the leukemic progenitor cell (LPC) burden in the pretreatment bone marrows of 15 pediatric T lineage ALL patients with an isolated extramedullary first relapse. Sites of extramedullary relapse were CNS (11 patients), testes (3 patients), and both CNS and testes (1 patient). Bone marrow LPC were detectable in 8 patients (53%) and undetectable in 7 patients (47%) at day 0 of post-relapse induction therapy, with LPC counts ranging from 0/10(6) mononuclear cells (MNC) to 518/10(6) MNC (mean +/ SEM, 50+/-34/10(6) MNC). Five of 9 patients with an early relapse (< 18 months after achieving a first complete remission [CR1]) and 3 of 6 patients with a late relapse (> or = 18 months from CR1) had detectable bone marrow LPC at day 0. Five of 8 patients with NCI-defined poor risk ALL and 3 of 7 patients with NCI-defined standard risk ALL had detectable LPC at day 0. Following post-relapse induction chemotherapy. LPC counts were detectable in bone marrows of 4 of 6 evaluated patients. Thus, approximately half of the extramedullary relapse T-lineage ALL patients studied had substantial occult involvement of the bone marrow. These findings may partly explain the previously observed poor prognosis of T-lineage patients following a CNS relapse. PMID- 11426550 TI - The combination of zidovudine and interferon alpha-2B in the treatment of adult T cell leukemia/lymphoma. AB - Adult T-cell leukemia/lymphoma (ATL) is frequently a very aggressive malignancy with a poor survival despite aggressive multiagent chemotherapy. The combination of the antiretroviral drug zidovudine (AZT) and interferon alpha (IFNalpha) has been reported to induce remissions in patients with ATL. The purpose of this study was to evaluate the clinical response and toxicity following administration of a combination of IFNalpha-2b and AZT in patients with human T-cell lymphotropic virus type I (HTLV-I)-associated ATL. Eighteen patients with ATL (chronic. crisis, acute or lymphoma type) were treated with the combination of AZT (50 - 200 mg orally 5 times a day) and IFNalpha-2b (2.5 - 10 million units subcutaneously daily). Three patients had objective responses lasting more than one month. One patient had a clinical complete remission, lasting 21.6 months and two patients had partial remissions lasting 3.7 and 26.5 months. Six patients were not considered evaluable for response due to short and/or interrupted periods of treatment. Seventeen patients have died with a median survival time after initiation of therapy of 6 months. Neutropenia and thrombocytopenia were the dose limiting toxicities. In conclusion, the response rate in this study was lower than noted in the two previous published series. This may be due to the amount and type of prior treatment our patients had received. PMID- 11426551 TI - Treatment of "poor risk" acute myeloid leukemia with fludarabine, cytarabine and G-CSF (flag regimen): a single center study. AB - We describe a single center experience of 41 consecutive patients with poor prognosis acute myeloid leukemia (AML) who received a single course of FLAG regimen consisting of Fludarabine 30 mg/m2/day plus Cytarabine 2 gr/m2/day (days 1-5) and G-CSF 5 mg/Kg/day (from day 0 to polymorphonuclear recovery) as salvage therapy. Eleven patients were primarily refractory to previous chemotherapy, 10 patients were in first relapse, 2 patients in second relapse and 7 patients in relapse after transplants. Eleven cases were defined as secondary AML (diagnosis of AML made after a preexisting diagnosis of myelodysplastic syndrome). The median age was 52.6 years (range 16-72); 29 patients were males and 12 females. Overall, 23 (56%) patients reached complete remission (CR), 3 patients died of infection (2) or hemorrhage (1) during induction, and 15 (36%) patients had resistant disease. The highest CR rates (80%) were obtained in relapsed cases; de novo and secondary AML registered 60% and 45% of CR rates, respectively. Patients achieving CR received a second FLAG course as consolidation and were submitted to an individualized program post-remission therapy, depending on the age and performance status. Hematological and non hematological toxicities were acceptable. In conclusion, our data confirm that FLAG is a an high effective treatment for poor prognosis AML and in young patients allows intensive post remissional therapy including allogeneic BMT. PMID- 11426552 TI - First line therapy with fludarabine combinations in 42 patients with either post myelodysplastic syndrome or therapy related acute myeloid leukaemia. AB - Acute myeloid leukaemias (AML) evolving from a myelodysplastic syndrome (MDS) or secondary to chemoradiotherapy frequently display unfavorable biologic characteristics. This may explain the lower remission rate obtained with conventional chemotherapy. Recently, the association of Fludarabine with intermediate dose Ara-C has produced interesting results particularly in high risk AML patients. Here, we report on 42 secondary AML patients treated with a combination of Fludarabine, intermediate dose Ara-C, G-CSF with or without an antracycline (FLANG, FLAG-IDA or FLAG). Overall, complete remissions (CR) were documented in 14 patients (33%) and partial responses (PR) in 12 (29%), while 10 patients proved resistant (24%). Six patients (14%) died early. The presence of a prognostically unfavorable karyotype had a negative impact on the CR rate (20% compared to 50% for patients with an intermediate prognosis karyotype, p 0.05). Patients treated with FLAG, FLANG and FLAG-IDA had similar CR rates. At the time of this analysis, after a mean follow-up of 12 months, the mean duration of CR is 16 months (range 3-66) and the mean survival is 11 months (range 1-67). The median time to granulocyte recovery (neutrophils > 0.5 x 10(9)/l) was 20 days (range 12-39) and 50 x 10(9)/l platelets were reached at a median of 26 days (range 9-56). Taken together, these Fludarabine containing regimens proved to be an effective and tolerable treatment for patients with secondary AML. Patients above 70 years of age may also benefit from this therapy, however the problem of treating patients with adverse chromosomal abnormalities still remains unresolved. PMID- 11426553 TI - Mitoxantrone and fludarabine in the treatment of patients with non-Hodgkin's lymphoma failing primary therapy with a doxorubicinor mitoxantrone-containing regimen. AB - Patients with recurrent lymphoma of any grade were treated with mitoxantrone (12 mg/m2 given intravenously (IV) over 15-30 minutes on day 1) followed by fludarabine at a dose of (25 mg/m 2 given IV over 30 minutes on days 1-3) every 28 days fludarabine at a dose of (25 mg/m2 given IV over 30 minutes on days 1-3) every 28 days. All patients had failed one prior chemotherapy regimen that contained either doxorubicin or mitoxantrone, total dose not exceeding 350 mg/m2 doxorubicin or 80 mg/m2 mitoxantrone. mitoxantrone. Thirty one patients (22 with intermediate- or high-grade and 9 with low-grade NHL) were enrolled. Median age was 63 years (range: 21 to 87). The objective response rate for patients with intermediate/high-grade NHL was 55% (27% with CR) and 89% (56% with CR) for patients with low-grade NHL. Median time to disease progression was 5.1 months for patients with intermediate/high-grade NHL and 10.8 months for patients with low-grade NHL. Median time to death for patients with intermediate/high-grade disease was 11.4 months. Median time to death for patients with low-grade NHL was not calculable as only one death (due to respiratory failure) occurred in this group 6.5 months after study start. The regimen was well tolerated. Grade 3/4 neutropenia was reported in 80% (24 of 30) of patients and Grade 3/4 thrombocytopenia in 19% (6 of 31) of patients. Nine hospitalizations for adverse events (primarily fever and neutropenia) occurred among eight patients, all with intermediate/high-grade NHL, during a total of 118 cycles of therapy. Further studies of this combination regimen in patients with intermediate/high-grade NHL and studies combined with monoclonal antibodies in low-grade NHL are warranted. PMID- 11426554 TI - Effectiveness of interferon-alfa and mid-cycle chemotherapy added to an anthracycline-based regimen in the treatment of aggressive non-Hodgkin's lymphoma. AB - Interferon-alfa in combination with cytotoxic chemotherapy has been shown to be effective in treating certain types of non-Hodgkin's lymphoma (NHL) (1). However, there is no published data on upfront induction treatment of aggressive NHL with IFN-alfa containing regimens. Studies have also shown that one can overcome regrowth resistance by administering mid-cycle agents which slow tumor proliferation between courses of cytotoxic therapy (2). Based on this, we treated 32 consecutive patients between 1/93 and 9/96 with a regimen containing cyclophosphamide 750 mg/m2, mitoxantrone 12 mg/m2, and teniposide 60 mg/m2 IV on day 1 with prednisone 100 mg PO given on days 1-5. On day 15, patients received vincristine 1.4 mg/m2 (2 mg max.) and bleomycin 10 units/m2 IV. Interferon-alfa 2b 5x10(6) units/m2 SQ was administered on days 22-26. The median age was 55 (range 26-83), M:F ratio was 2.5:1, and the median International Prognostic Index was 2. 38% of patients had stages I-II and 62% had stages III-IV disease. Fifty nine percent of the patients achieved a complete response, 22% a partial response, and 19% had progressive disease. The overall survival (OS) was 81% and the progression free survival (PFS) was 56% at 4.3 years. There were no severe (grade IV) hematologic, flu-like, GI and infectious toxicities from IFN-alpha. Leukopenia was the main severe toxicity related to the chemotherapy regimen (days 1-15), but not IFN-alpha. Severe infection secondary to the chemotherapy regimen occurred in one patient. Interferon-alfa-2b and mid-cycle chemotherapy added to an anthracycline based regimen is effective induction treatment for patients with aggressive NHL. The OS and PFS using this regimen, based on regrowth resistance, appears to be at least as or more effective than CHOP therapy for this group of patients. Severe toxicities were rare. PMID- 11426555 TI - Fludarabine, arabinosyl cytosine and idarubicin (FLAI) for remission induction in poor-risk acute myeloid leukemia. AB - Progress in treatment of acute myeloid leukemia (AML) is slow and treatment intensification alone has limited effects, particularly in poor-risk cases. Poor risk cases, that are identified mainly by prior history, leukemic cell mass and cytogenetic abnormalities, share multiple mechanisms of drug resistance that are responsible for treatment failure. Since Pgp-mediated resistance to anthracycline can be reduced with Idarubicin (IDA) and resistance to arabinosyl cytosine (AC) can be reduced with Fludarabine (FLUDA), we tested a combination of high dose AC (2000 mg/sqm, 5 doses), FLUDA (30 mg/sqm, 5 doses) and IDA (12 mg/sqm, 3 doses) for remission induction and consolidation in 45 consecutive cases of poor-risk AML. The complete remission (CR) rate was 71% after the first course and 82% overall, with a projected 2-year survival and relapse-free survival of 44% and 50% respectively. Non-hematologic toxicity was very mild, that is very important in elderly patients, but hemopoietic toxicity was substantial, with a time to hematologic recovery of 3 to 4 weeks and two cases of death in CR. Peripheral blood stem cells (PBSC) could be mobilized and collected successfully only in 11 cases. This three-drug combination is effective and has a limited non-hematologic toxicity, but FLUDA may increase the difficulty of obtaining PBSC early after remission induction. PMID- 11426556 TI - Amifostine in combination with erythropoietin and G-CSF promotes multilineage hematopoiesis in patients with myelodysplastic syndrome. AB - Ineffective hematopoiesis leading to profound cytopenias represents a major clinical problem in the management of patients with myelodysplastic syndrome (MDS). The aminothiol amifostine has shown to promote multilineage hematopoiesis both in vivo and in vitro in patients with MDS. We have treated 10 patients with 250 mg/m2 amifostine thrice weekly in combination with erythropoietin for 4 consecutive weeks followed by 2 weeks observation. Responding patients received the same 6 week schedule, while nonresponder received G-CSF in addition to erythropoietin and amifostine during the second treatment course. All patients experienced single or multilineage hematologic improvement, but only 2 reached transfusion independency. Moreover, response was durable only in a minority of patients and thus additional studies are warranted to further define the potential interaction of amifostine and growth factors. PMID- 11426557 TI - Lack of correlation between clinical characteristics and serum soluble Fas ligand levels in patients with multiple myeloma. AB - Multiple myeloma is characterized by the accumulation of malignant plasma cells in the bone marrow and rarely cured by chemotherapy. Villunger et al. showed that the neoplastic plasma cells express Fas ligand (FasL), which transmits a signal of apoptosis upon ligation to Fas, and suggested that the FasL suppresses the T cells activated against malignant cells, resulting in escape from tumour immunity. We examined serum soluble FasL (sFasL) levels in 35 multiple myeloma patients to evaluate the correlation between sFasL levels and clinical characteristics. The serum sFasL levels were not affected by the disease status, serum monoclonal protein levels, or other prognostic factors. We could not determine whether the expression of FasL is involved in the poor clinical course of the disease. PMID- 11426558 TI - Bleomycin, lomustine, cyclophosphamide, vincristine, procarbazine and prednisone (BLEO-CCVPP) in patients with Hodgkin's disease who relapsed after radiotherapy alone: a long-term follow-up study of the Eastern Cooperative Oncology Group (E3481). AB - Thirty-three evaluable patients with Hodgkin's disease who failed radiotherapy were treated on this phase II study with bleomycin, lomustine, cyclophosphamide, vincristine, procarbazine and prednisone given every 28 days for a minimum of eight courses. Twenty-five patients (76%; 95% CI=55.6-87.1%) achieved a complete remission, the median duration of which cannot yet be determined, but the probability of remaining in continuous complete remission at 10 years is.64. The median survival from entry on this study for all evaluable patients is 10 years, and 12 patients were alive at the time of this analysis with a median follow-up for them of 15.5 years. Of the 22 patients who died, 11 died of progressive or recurrent Hodgkin's disease and 11 died of other causes including 7 second primary neoplasms and at least one myocardial infarction. Both are now well known late complications of Hodgkin's disease treatment. PMID- 11426559 TI - Vascular access via peripherally inserted central venous catheters (PICCs): experience in 40 patients with acute myeloid leukemia at a single institute. AB - Reliable long-term vascular access is essential for the treatment of patients with acute myeloid leukemia (AML). Although peripherally inserted central catheters (PICCs) have been in use for many years, little data exist on their use in patients receiving intensive chemotherapy. We retrospectively reviewed all AML patients who had a PICC inserted between July 95 and May 98. Fifty two PICCs were inserted in 40 patients with AML. Thirty three PICCs were inserted during severe thrombocytopenia (platelets < 50 x 10(9)/L), and 31 during severe neutropenia (neutrophils < 0.5 x 10(9)/L). Mean catheter duration was 82 (median 63, range 3 441) days for a total of 4274 catheter days. A mean of 1.8 chemotherapy courses were administered via each PICC. There were 5 early complications of PICC placement. Other mechanical complications occurred in 14 catheters and phlebitis in 12. Twenty blood stream infections (BSI) occurred in 17 patients. All BSIs occurred during neutropenia. Seventeen PICCs were removed due to the following complications - phlebitis (11), possible catheter related BSI (4), mechanical reasons in 3 (2 with concomitant phlebitis) and persistent fever (1). PICC duration was significantly shorter in these 17 catheters (52.9 v 96.4 days in the other 35, p=0.0289). We conclude that PICCs provide long-term vascular access with an acceptable complication rate in patients with AML. However, a randomised trial is required before PICCs can be considered an alternative to tunneled central venous catheters in these patients. PMID- 11426560 TI - Tumour kinetics in multiple myeloma before, during, and after treatment. AB - Tumour progression was monitored in seven multiple myeloma (MM) patients undergoing a novel oral chemotherapy regimen (cyclophosphamide, idarubicin and dexamethasone; CID) followed by early autologous stem cell transplantation (ASCT). Allele-specific oligonucleotide PCR (ASO-PCR) was used to semi-quantitate the number of tumour cells within the peripheral blood (PB) and PB progenitor cell (PBPC) harvests and compared with paraprotein levels and morphological bone marrow (BM) assessments. Tumour cells were detected in the PB of all patients at diagnosis, but decreased in response to CID therapy. All but two of the 22 PBPC collections contained MM cells, the levels of which were statistically correlated with overall clinical response to therapy, but not with individual BM or PB tumour loads prior to mobilisation. We also found no correlation between the day of leucapheresis collection and the number of contaminating MM cells, CD34+ cells or MM cells per CD34+ cell. Regardless of tumour contamination levels in the PBPC collections, the majority of patients demonstrated post-ASCT clearing of circulating MM cells. This study suggests that levels of circulating MM cells may be the best indication of patient response to treatment and argues against the theory of differential mobilisation of tumour cells and CD34+ cells in response to cytokine treatment. PMID- 11426561 TI - Prevention of acute graft-versus-host-disease by selective depletion of T cells reactive with minor histocompatibility antigens on epithelial cells. AB - Graft-versus-host disease (GVHD) is a major obstacle in allogeneic hematopoietic stem cell transplantation (HSCT). Mature donor T-cells present in the graft play a pivotal role in the development of acute GVHD. On the other hand, mature donor T-cells in the graft are also crucial for the elimination of residual tumor cells still present in the patient after HSCT. Whether donor T cells act non specifically against the patient, including an overlapping GVHD/GVL reactivity, or some donor T cells have GVHD reactivity while other donor T cells have GVL reactivity is still unclear. Some in-vitro data are suggestive that selective T cell depletion techniques are possible by which GVHD-reactive T cells can be eliminated while GVL-reactive T cells are preserved. Here we update some approaches of selective T cell depletion that have been developed in our laboratory. PMID- 11426562 TI - Gamma-linolenic acid induces apoptosis in B-chronic lymphocytic leukaemia cells in vitro. AB - Gamma linolenic acid (GLA) is cytotoxic to many types of human cancer cells. Most chemotherapeutic agents are cytotoxic by inducing apoptosis. We examined the apoptotic activity of GLA on purified B-cells isolated from patients with B-cell chronic lymphocytic leukaemia (B-CLL) and from normal individuals. GLA significantly increased the degree of apoptosis in B-CLL B-cells after 24 hours of culture. The mean percentage of cells undergoing apoptosis when cultured in medium alone (spontaneous apoptosis) was 20% (range: 7 to 31%) (n=25) and in the presence of GLA (5 microg-60 microg) was: 42%-95%. In the presence of GLA 5 microg/ml and dexamethasone the degree of apoptosis was 86% (range: 72 to 100%). GLA induced apoptosis in B-CLL B-cells at a higher level than that observed with normal B-cells at all lower concentrations tested 5, 10 and 15 microg/ml: P=0.045; 0.027 and 0.022, respectively. At 30 microg/ml of GLA, no significant difference in the percentage of cells displaying apoptosis between B-CLL and normal B-cells was observed (P=0.075). GLA induced apoptosis in B-CLL T-cells at both 10 and 30 microg/ml. The degree of apoptosis in normal T-cells with GLA was also significant at the higher concentration of 30 microg/ml. Interleukin 4 (IL4), a viability factor in B-CLL, and vitamin E, an anti-oxidant, protected B CLL B-cells against GLA (20 microg/ml)-induced apoptosis. These results demonstrate that GLA induces apoptosis in B-CLL B- and T-cells cells in vitro and that they are more susceptible to GLA-induced apoptosis than normal peripheral blood B- and T-cells. PMID- 11426563 TI - Expression of EBV encoded nuclear small non-polyadenylated RNA (EBER) molecules in 32 cases of childhood Burkitt's lymphoma from Israel. AB - We have analyzed paraffin sections from 32 children with histologically confirmed Burkitt's Lymphoma (BL) for the presence of EBV using in situ hybridization to detect expression of the EBV-encoded early RNAs (EBERs). EBV was present in the tumors of 11 patients (34%). Sixty nine percent of the children presented with abdominal disease, 19% had bone marrow infiltration and only one child had jaw involvement. There was no statistically significant difference between EBV positive and EBV negative children with regard to age, gender, origin, primary site at presentation, or clinical stage of disease. However, there was a trend for younger age in the children with EBV positive BL with a median age of 4, compared to 7 years in children with EBV negative BL. None of the 7 children of Ashkenazi Jewish origin had EBER positive disease. There was no difference in the treatment outcome between the EBV positive patients (estimated survival at 24 months of 82%) and EBV negative children (estimated survival rate of 71% (p=0.58)). In conclusion, although this is only a small series it seems that childhood BL in Israel has the clinical characteristics of sporadic, non-African type with 34% EBV association and a low incidence of jaw tumors. Our data suggest that Ashkenazi Jewish children with BL are less likely to have EBV positive tumors than other ethnic groups. However, more patients will need to be studied in order to assess the validity of this observation. PMID- 11426564 TI - Absence of mutations in the CDKN2 binding site of CDK4 in childhood acute lymphoblastic leukemia. AB - The cell cycle regulatory circuit resulting in phosphorylation of the retinoblastoma protein (pRB) is frequently altered in human cancers. Several mechanisms of disruption are known in that pathway. In childhood acute lymphoblastic leukemia (ALL), the main disrupting mechanism is the homozygous deletion of the CDKN2 (cyclin dependent kinase inhibitor 2) genes: p16CDKN2a, p15CDKN2b, and p19ARF. Another pRB pathway disturbance is a previously described point mutation in the exon 2 of CDK4, a pRB phosphorylating enzyme, which abrogates binding of the latter to its inhibitors, p16CDKN2a and p15CDKN2b. Here we report the absence of point mutations in the CDKN2-binding site of CDK4 in 100 cases of childhood ALL, 2 cases of childhood chronic myeloid leukemia and 9 hematologic cell lines screened by PCR-SSCP (polymerase chain reaction single stranded conformational polymorphism gel electrophoresis), thereby minimizing the possibility of the existence of these specific CDK4 mutations in childhood ALL. PMID- 11426565 TI - Establishment of a human cell line (SKI-DLCL-1) with a t(1;14)(q21;q32) translocation from the ascites of a patient with diffuse large cell lymphoma. AB - Cytogenetic abnormalities at chromosome 1q21 are among the most common second genetic events observed in Non-Hodgkin's Lymphomas and have prognostic significance. Recently, BCL9 has been cloned from a pre-B-cell lymphoblastic leukemia cell line, which carried a t(1:14)(q21;q32). However, among a panel of 39 B-cell malignancies with 1q21 translocation, only two cases showed rearrangement for the BCL9 gene. We report the establishment of a new lymphoma cell line from a patient with relapsed diffuse large cell lymphoma. This cell line SKI-DLCL-1 showed cell surface antigens identical to the original tumor and demonstrated the profile of a mature B-cell phenotype: CD19 and CD20 positive, CD5 and C10 negative. It carried a t(1;14)(q21;q32) translocation identical to the original tumor. Although the clinical presentation was an isolated effusion lymphoma, studies for HIV-1, HHV8 and EBV were all negative. Southern blot analysis demonstrated that BCL9 was not rearranged in the SKI-DLCL-1 cell line. In addition, the BCL9 gene was not over-expressed in SKI-DLCL-1 cell line. The identification of a new locus at 1q21 will help clarify the pathogenesis of B cell malignancies with a translocation involving this locus. PMID- 11426566 TI - Nodal plasmacytoma with significant paraproteinaemia. AB - We present a case of primary nodal plasmacytoma in an elderly Chinese woman that was associated with significant paraproteinaemia and paraproteinuria. Clinical and laboratory features of the patient satisfied Durie's criteria for the diagnosis of multiple myeloma. The present case was unusual in two aspects. Firstly, there was no evidence of clonal plasma cell proliferation elsewhere in the body after extensive radiological investigations, repeated bone marrow examinations, and polymerase chain reaction for immunoglobulin gene rearrangement study. Secondly, the clinical behaviour was indolent despite the large amount of paraprotein production, and showed satisfactory disease control with local radiotherapy. The differential diagnoses of plasmacytosis in the lymph node are also discussed. PMID- 11426567 TI - Cryoprecipitate-induced mesenteric venous thrombosis during L-asparaginase therapy for acute lymphoblastic leukaemia. AB - We present a case of fatal mesenteric vein thrombosis (MVT) associated with L asparaginase (L-asp) therapy and temporally related to cryoprecipitate infusion, in an adult with acute lymphoblastic leukaemia (ALL). Cryoprecipitate was given on two consecutive days to raise a low fibrinogen level of 0.7 g/L, in the presence of severe thrombocytopenia and mucocutaneous bleeding. The thrombotic event presented as sudden abdominal pain a day after the second cryoprecipitate infusion, which raised the fibrinogen to 1.5 g/L. Concurrent levels of antithrombin III (AT III), protein C (PC) and protein S (PS) were very low. The patient died after laparotomy and wide resection of gangrenous bowel. We believe this is the first reported case in the English literature of a patient who developed mesenteric venous thrombosis during L-asp therapy, and once more we advise caution in using conventional blood products, especially cryoprecipitate, and recommend restricting the use of cryoprecipitate and fresh frozen plasma (FFP) to the treatment of serious hemorrhagic manifestations, until new effective and safe therapies are available. PMID- 11426568 TI - Leukemia cutis in an elderly patient treated with low dose cytosine arabinoside and etoposide. AB - A 78-year-old man presented with a generalized erythematous papular rash. Such skin lesions were not painful, tender or pruritic, and spread over the truncus. He was admitted to our hospital for examination of the skin lesions. Laboratory tests indicated microcytic hypochromatic anemia and thrombocytopenia, although gave a normal leukocyte count with normal differentiation. His bone marrow showed hypercellularity, with 43% peroxidase positive blasts that displayed positive immunophenotypes for CD4, CD13, CD33, CD41a, KP-1 (CD68), and HLA-DR. His skin specimen revealed infiltration in the dermis and subcutaneous fat tissue by leukemic cells that were positive for the leukocyte common antigen (LCA, CD45), CD15, CD33, CD68, and HLA-DR. He was diagnosed as having M4 subtype of acute myelogeneous leukemia (AML) with leukemia cutis. After three courses of low dose cytosine arabinoside (LDAC), combined with low dose etoposide, he achieved complete remission (CR). He remained well, with no evidence of relapse nine months later. LDAC should be considered as initial treatment for such cases of leukemia cutis with poor general condition. PMID- 11426569 TI - Subcutaneous extramedullary hematopoiesis in a patient with secondary myelofibrosis following polycythemia vera. AB - We describe the case of a 73-year-old woman with secondary myelofibrosis who developed subcutaneous extramedullary hematopoiesis. Although extramedullary hematopoiesis has been generally observed in primary myelofibrosis, in this case it was seen in myelofibrosis secondary to polycythemia vera. Histological examination of the subcutaneous nodule revealed that the lesion included cells from the myeloid and megakaryocytic series. The skin lesion almost disappeared after treatment with hydroxyurea. We report here this rare manifestation in secondary myelofibrosis including a review of literature. PMID- 11426570 TI - Systemic fusariosis after a preparative regimen including thiotepa, VP-16 and busulfan used for blood stem cell transplantation in Hodgkin's disease. AB - Fusarium infection is rare but important infection after bone marrow transplantation (BMT). A 27-year-old man developed systemic fusarial infection following severe skin damage probably caused by high-dose thiotepa administration. Systemic fusariosis rapidly progressed to a variety of organs despite antifungal treatment, and he finally died of this infection on day 75. Considering that this organism usually invades via damaged skin and that the penile lesion was the first manifestation of systemic fusariosis in this patient, careful examination of the skin might be helpful for early diagnosis of fusarial infection. His clinical course provided us with an important clue for diagnosis of fusarial infection. PMID- 11426571 TI - Incidental diagnosis of Gaucher's disease in an elderly patient with lymphoma. PMID- 11426572 TI - Mini-ice protocol is better than high-dose hydroxyurea to mobilize Ph-negative cells in earlier phases of chronic myelogenous leukemia. PMID- 11426573 TI - Chemokinetic behavior of the infective third-stage larvae of Strongyloides ratti on a sodium chloride gradient. AB - The movements of the infective third-stage larvae (L3) of a rodent parasitic nematode Strongyloides ratti were examined on a sodium chloride (NaCl) gradient set up on agarose plates. The movements of larvae were followed by observing their tracks on the surface of the agarose. The direction of movement depended on the NaCl concentration at the point of their initial placement on the gradient. Larvae placed at between 230 and 370 mM NaCl tended to migrate towards areas of lower concentration. On the other hand, when placed at concentrations less than 20 mM NaCl, larvae tended to migrate initially towards higher concentrations but did not linger in areas where the concentration was over approximately 80 mM NaCl. It seems that S. ratti L3, tested in vitro, prefer regions with a concentration of NaCl below 80 mM NaCl. Two typical chemokinetic behaviors are seen; a unidirectional avoidance movement when initially placed in unfavorable environmental conditions and a random dispersal movement when placed within an area of favorable conditions. Track patterns were straight in the avoidance movement but included multiple changes of direction and loops in the dispersal movement. This study introduces an assay system suitable for studying chemokinetic behavior of larvae of Strongyloides ratti. PMID- 11426574 TI - Laboratory maintenance of Eucyclops serrulatus (Copepoda: Cyclopoida). AB - A simple method for culturing a freshwater copepod Eucyclops serrulatus is described. A flagellate Chilomonas paramecium serves as the food organism. Starting from a single, egg-bearing female collected in a pond, the copepod proliferated for at least 10 generations apparently with no adverse changes. The stock culture has been maintained with minimal effort for at least 19 months. The method is expected to make the copepod useful in the laboratory as an intermediate host of various parasitic helminths. PMID- 11426575 TI - The 1992-1999 World Bank Schistosomiasis Research Initiative in China: outcome and perspectives. AB - Ongoing efforts over the last 50 years, aiming at the elimination of schistosomiasis in the People's Republic of China, have been spectaculary successful in reducing the prevalence and intensity of the infection. The endemic areas have been reduced to core regions with particular problems such as the middle and lower reaches of the Changjiang River (Yangtze), the land adjacent to the lakes of central China and certain mountainous areas in Sichuan and Yunnan. An effort to eradicate schistosomiasis as a public health problem in these areas, by means of mass chemotherapy in regions of high prevalence and selective chemotherapy in others, provided good results initially but a lasting effect proved unattainable with chemotherapy alone. A small part of the funds available for this effort were used for research and training. Overseen by a Joint Research Management Committee (JRMC), research training was intensified resulting in improved applications and a better quality of the scientific level of the research finally carried out. Several new control tools were produced which may improve future control approaches, which might achieve a more than temporary relief. In evaluating the contributions made, it was found that the great environmental variations between the eight provinces where control activities were implemented was the main reason why general use of chemotherapy only could not be entirely successful. The inclusion of a research component proved beneficial both for the short- and long-term control and the JRMC proved useful in exposing that sustained progress cannot be achieved without back-up by other approaches, e.g. snail control. Suggested future activities include strengthening of intersectoral and industrial collaboration but finding financial support for continuing the JRMC initiative in some form. It is crucial to consolidate progress made. PMID- 11426576 TI - Differential diagnosis of schistosomiasis mekongi and trichinellosis in human. AB - An indirect (plate) ELISA and, a more convenient version, a dot-blot (membrane) ELISA have been developed using haemocyanin of a mollusk, Megathura crenulata, i.e. keyhole limpet haemocyanin (KLH) and purified, specific antigen of Trichinella spiralis (APTsAg) obtained from a monoclonal antibody-affinity column chromatography, for differential diagnosis of schistosomiasis mekongi and trichinellosis. Serum samples of patients with parasitologically confirmed trichinellosis were reactive to both antigens in both versions of ELISA while sera of patients with schistosomiasis mekongi were positive only to the KLH. Both ELISA were negative when used to test sera of normal controls and patients with gnathostomiasis, paragonimiasis and opisthorchiasis. PMID- 11426577 TI - Serum factors governing intraerythrocytic development and cell cycle progression of Plasmodium falciparum. AB - Malaria is clinically manifested only when the human malaria parasites in the genus Plasmodium enter the obligatory intraerythrocytic life cycle. Elucidation for the roles of the serum, the key nutrient, and its components is then deemed essential for thorough understanding of the proliferation of Plasmodium cells at the erythrocytic stage. Fractionation and analysis of serum and its components was performed by chromatography, solvent extraction, and subsequent reconstitution experiments. Only fractions containing serum albumin (SA) from the serum and purified intact bovine serum albumin (BSA) showed comparable growth promoting activity with human serum (HS). Delipidated BSA can only effect parasite growth after reconstitution with lipids extracted from intact BSA. Fatty acid (FA) species in the neutral lipid fraction from intact BSA proved likewise when reconstituted with delipidated BSA. Furthermore, the involved FA species have to come in a pair of one saturated and one unsaturated, with palmitic and oleic acids as the best combination. The results were further substantiated by morphological analysis as well as biochemical analysis of the DNA synthesis during the intraerythrocytic development. This study can be a basis to explore the molecular mechanism of lipid traffic within the parasitized red blood cell (RBC), which can be an important adjunct to the development of drugs for malaria therapy. PMID- 11426578 TI - Molecular evidence of natural hybridization between Fasciola hepatica and F. gigantica. AB - Nucleotide sequences of two regions, cytochrome c-oxidase subunit 1 (CO1) and NADH dehydrogenase subunit 1 (ND1) of the mitochondrial DNA and two regions, internal-transcribed spacer 2 (ITS2) and the D2 region in the 28S rDNA (28S) of the nuclear DNA were obtained from five Korean worms of the genus Fasciola in order to elucidate their taxonomic status. The CO1 and ND1 regions are all monomorphic in the Korean worms and similar to those of F. gigantica. On the other hand, the ITS2 and D2 regions were found to be polymorphic; that is, out of five worms, two possessed a F. gigantica-type sequence, one, a F. hepatica-type sequence and two possessed sequences of both types indicating an existence of different alleles at the loci. It should be noted that these variations of the ITS2 and D2 regions co-occur at the same individual worms. This was confirmed by sequencing five to six cloned PCR products for each worm. The present study strongly suggests interspecific cross-hybridization between the two species coexisting in Korea. PMID- 11426579 TI - Ultrastructural and immunohistochemical analysis of cholangiocarcinoma in immunized Syrian golden hamsters infected with Opisthorchis viverrini and administered with dimethylnitrosamine. AB - Utilizing the experimental model in Syrian golden hamsters, we explored the role of immunization in carcinogenesis. The animals, which were infected with liver flukes (Opisthorchis viverrini), and administered a subcarcinogenic dose of dimethylnitrosamine, developed cancer. Pre-immunizing with a crude somatic antigen did not reduce cancer development, but accelerated carcinogenesis. Histopathological analysis of the cancer tissues was done once at week 30 and again at week 39 using H and E staining, immunostaining for the p53 tumor suppressor phosphoprotein, and electron microscopy. Thirty weeks after immunization, the immunized hamsters developed tubular adenocarcinoma at a higher rate (71.43%) than the non-immunized group (20.00%). This rate (20.00%) increased to 63.64% by week 39. The small foci cancer in the non-immunized group decreased in frequency from 80.00% (at week 30) to 36.36% (by week 39), suggesting the small foci cancer progressed to tubular adenocarcinoma during the 9-week interval. Most of the observed tubular adenocarcinoma was well differentiated. Nearly all hamsters that tested positive for cancer also tested positive for p53 immunostaining in the epithelia of the small bile ducts. The positive reaction for p53-immunostaining was localized in the rough endoplasmic reticulum, Golgi apparatus and perinuclear membranes. The electron micrographs of these positive p53-immunostained cells showed characteristics of early cancer. The detection of p53 in early cancer development makes it a candidate as a tumor marker. PMID- 11426580 TI - Prevalence of dirofilarial infection in raccoon dogs in Japan. AB - The raccoon dog (Nyctereutes procyonoides viverrinus) is known to acquire canine heartworm (Dirofilaria immitis) infection. We surveyed the prevalence of heartworm infection in free-ranging raccoon dogs in the Nishi-Tama (Tokyo) and Kanagawa areas of Japan. A total of 75 raccoon dog carcasses, including 29 animals from the Nishi-Tama area and 46 from the Kanagawa area, were necropsied between 1992 and 1993. Eight out of 75 raccoon dogs were found to be infected (overall 10.7%). The prevalence of infection was 6 and 16% in Nishi-Tama and Kanagawa, respectively. Microfilarial production was observed in the uterus of one female adult dog. PMID- 11426581 TI - Effects of gliclazide on low-density lipoprotein oxidizability and atherosclerosis in cholesterol-fed rabbits. AB - We studied the effects of a widely-used sulfonylurea, gliclazide, on the oxidizability of low density lipoprotein (LDL) and the development of experimental atherosclerosis in cholesterol-fed rabbits. Daily oral administration of gliclazide (20 mg/kg/day) tended to inhibit the aortic atherosclerosis induced by feeding a 1% cholesterol diet for 10 weeks, although it did not affect diet-induced hyperlipidemia. The administration of gliclazide tended to inhibit the increase of serum thiobarbituric acid-reacting substances (TBARS) by cholesterol feeding and to increase the lag time of the conjugated diene formation of LDL subjected to in vitro oxidation by copper ion, although without significance. The present study suggests that gliclazide may have antioxidative properties in vivo, and have further beneficial effects for the treatment of diabetes mellitus by inhibiting the oxidation of LDL. PMID- 11426582 TI - Pravastatin use and risk of coronary events and cerebral infarction in japanese men with moderate hypercholesterolemia: the Kyushu Lipid Intervention Study. AB - The Kyushu Lipid Intervention Study (KLIS) aimed to investigate the effects of pravastatin in the primary prevention of coronary events and cerebral infarction in Japanese men aged 45-74 years with serum total cholesterol of > or = 220 mg/dl (5.69 mmol/l). The coronary events included myocardial infarction, coronary artery surgery and angioplasty, cardiac death, and sudden death. A total of 5,640 patients were recruited, and 3,061 and 2,579 were allocated to the pravastatin (10-20 mg/day) and conventional treatment, respectively. They were followed up for 5 years on average. Due to unsuccessful randomization, a protocol-based analysis was carried out with adjustment for coronary risk factors at baseline using the Cox proportional hazards model. For several reasons (serum total cholesterol of > or = 300 mg/dl, use of drugs excluded in protocol, etc.), 2,219 men in the pravastain group and 1,634 in the conventional treatment group were used in the analysis. The baseline total cholesterol levels were 254 mg/dl in the pravastain group and 243 mg/dl in the conventional treatment group. Serum total cholesterol levels fell by 15% in the pravastain group and by 8% with the conventional treatment group. Adjusted relative risks for pravastatin versus conventional treatment were: coronary events 0.86 (one-side p=0.23), cerebral infarction 0.78 (p=0.13), and the two outcomes combined 0.81 (p=0.08). These findings add to evidence that pravastatin use is beneficial in the prevention of coronary events and cerebral infarction in hypercholesterolemic Japanese patients, and suggest that pravastatin may be more protective against cerebral infarction. PMID- 11426583 TI - In vivo metabolism of HDL, apo A-I, and lp A-I, and function of HDL--a clinical perspective. AB - Serum levels of high density lipoprotein cholesterol (HDL-C) are inversely correlated with coronary heart disease (CHD). Kinetic studies indicate that the mechanism for the variation in HDL levels associated with various pathophysiologic states includes changes in the fractional catabolic rate (FCR) and/or the synthesis rate of HDL and its major proteins apolipoprotein (apo) A-I and apo A-II. The antiatherogenic effects of HDL are thought to be mainly due to its role in reverse cholesterol transport. HDL is an assembly of heterogeneous particles. HDL enlarges when it takes up cellular cholesterol, and shrinks when HDL cholesterol ester (CE) is transfered to low density lipoprotein (LDL) and very low density lipoprotein (VLDL) particles. The functional ability of HDL (to remove cellular cholesterol) has drawn considerable attention. The fractional esterification rate of cholesterol in HDL (FER(HDL)) has been established as a functional assay of HDL, and reflects the size of HDL particles. We investigated the clinical significance of FER(HDL) and its relationship to the quantity of HDL. FER(HDL) values were inversely correlated with levels of HDL-C and large lipoprotein containing apo A-I (LpA-I). The association between FER(HDL) and CHD changed with serum HDL-C levels: increased FER(HDL) values significantly increased the risk of CHD when serum HDL-C levels were low, while there was no such relationship when HDL-C levels were high. We concluded that the combination of HDL-C levels and FER(HDL) is a stronger indicator of CHD than either the HDL-C level (quantitative measure of HDL) or FER(HDL) (functional measure of HDL) alone. PMID- 11426584 TI - Structural and functional properties of apolipoprotein A-I mutants. AB - Apolipoprotein (apo) A-I is composed of 243 amino acid residues that fold into amphipathic helixes, and plays a central role in the high density lipoprotein (HDL) metabolism. Familial apoA-I deficiency is a rare metabolic disorder of which three cases have been characterized at a molecular level in western Japan. However, in subjects with apoA-I deficiency, coronary artery disease was not always present. One apo A-I deficiency was compound heterozygous apoA-I mutant for a TATA box mutation and a structural nonsense mutation. To date, screening analysis in our laboratory has identified nine genetically-determined structural mutations of apo A-I. We have also characterized these apo A-I mutations, including apoA-I (Glu235del) Nichinan. Few structural mutations were associated with altered HDL cholesterol levels. PMID- 11426585 TI - Therapeutic angiogenesis induced by hepatocyte growth factor: potential gene therapy for ischemic diseases. AB - Recent progress in molecular biology has led to the development of gene therapy as a new strategy to treat a variety of cardiovascular diseases. Targeted diseases range from single gene deficiency diseases to more complex diseases in adults such as restenosis after angioplasty. One obvious major target in the field of gene therapy is ischemic diseases such as myocardial infarction, angina and peripheral arterial diseases (i.e. ASO (arteriosclerosis obliterans)). In a large proportion of such patients, the anatomical extent and the distribution of arterial occlusive disease make the patients unsuitable for operative or percutaneous revascularization. Thus, the disease frequently follows an inexorable downhill course. Of importance, there is no optimal medical therapy for severe ischemic hearts and critical ischemic limbs. Therefore, novel therapies are required to treat these patients. Recently, the efficacy of therapeutic angiogenesis using VEGF (vascular endothelial growth factor) gene transfer has been reported in human patients with critical limb ischemia and myocardial ischemia. Thus, the strategy for therapeutic angiogenesis using angiogenic growth factors should be considered for the treatment of patients with critical limb ischemia or myocardial infarction. The endothelial cell specificity of VEGF has been considered to be an important advantage for therapeutic angiogenesis, as endothelial cells represent the critical cellular element responsible for new vessel formation. Indeed, human gene therapy for ASO and angina has already begun in the USA, with surprising and beneficial effects. We have focused on hepatocyte growth factor (HGF), which is a mesenchyme-derived pleiotropic factor that regulates cell growth, cell motility, and morphogenesis in various types of cells. Recently, HGF is also considered to be a powertul growth tactor for endothelial cells. In this review, we described the potential gene therapy for ischemic diseases using HGF. PMID- 11426586 TI - Troglitazone, a PPARgamma ligand, inhibits osteopontin gene expression in human monocytes/macrophage THP-1 cells. AB - Peroxizome proliferator-activated receptor-gamma (PPARgamma) is a member of the nuclear receptor family of transcription factors that regulate adipocyte differentiation. Recent studies indicate that liganded PPARgamma not only promotes differentiation but also inhibits the activation of macrophages. Osteopontin, a component of extracellular matrix, is synthesized by macrophages in atherosclerotic plaques. In this study, we examined whether PPARgamma ligand regulates osteopontin gene expression in THP-1 cells, a cell line derived from human monocytic leukemia cells which can differentiate to macrophage upon stimulation with phorbol ester PMA. Northern blot analysis showed that osteopontin expression is markedly induced in response to PMA. Troglitazone, a PPARgamma ligand, dramatically attenuated the PMA-induced osteopontin expression. Transient transfection assays of the human osteopontin promoter/luciferase construct which contains a 5'-flanking region between -1500 and +87 relative to the transcription start site demonstrate that either treatment with troglitazone or cotransfection of PPARgamma expression vector inhibits osteopontin promoter activity. These data indicate that troglitazone reduces osteopontin gene expression at transcriptional level through PPARgamma activation, and suggest the role of troglitazone in inhibiting the ability of macrophages to produce extracellular matrix, which is particularly relevant to atherosclerotic plaque formation. PMID- 11426587 TI - Atherosclerotic plaques composed of a large core of foam cells covered with thin fibrous caps in twice-injured carotid arterial specimens obtained from high cholesterol diet-fed rabbits. AB - We attempted to find atherosclerotic plaques including a large lipid core and thin fibrous cap in twice-injured arterial specimens obtained from high cholesterol diet (HCD) fed rabbits. Rabbits fed a HCD were subjected to carotid artery injury using a balloon catheter. After 2 or 4 weeks of cholesterol feeding, a second mild injury was induced in the same region as the first injury. The rabbits were given a standard diet for 2 weeks after the second injury. Typical atherosclerotic plaques with a fibrous cap formed by smooth muscle cells and extracellular matrix overlying a core formed by macrophage foam cells were observed in the lesion. Gelatin proteolytic activities were found in homogenates containing either media or intima from the injured artery, and activated matrix metalloproteinase-2 (MMP2) was detected. With prolongation of the HCD feeding period (interval between injuries) from 2 weeks to 4 weeks, typical plaque was observed more frequently. Furthermore, the neointimal area and the macrophage foam cells area increased, as did gelatin-proteolytic activity. Since the typical atherosclerotic plaques observed in the present study have some histopathological and pathogenic characteristics in common with unstable atherosclerotic plaque, we expect that the typical atherosclerotic plaque found in the present study will be useful for basic studies of plaque stabilization and prevention of acute coronary syndromes. PMID- 11426588 TI - Effect of bezafibrate or pravastatin on serum lipid levels and albuminuria in NIDDM patients. AB - Lipid abnormalities in diabetic patients, particularly in those with nephropathy, may be partially due to deteriorating atherosclerosis. Therefore, strict control of the lipid metabolism in addition to glycemic control is desirable. Whether or not lipid lowering drugs prevent albuminuria in diabetic patients in the long term remains unclear. This study involved 71 NIDDM patients with hypercholesterolemia (group A: n = 37, group B: N = 34). The effect of bezafibrate (group A) or pravastatin (group B) on the cholesterol (CH) content of apolipoprotein AI, B100 containing particles or remnant-like particles (RLP) or urinary albumin excretion was studied over 4 years. The CH content in apolipoprotein B100 particles after treatment with either bezafibrate or pravastatin decreased significantly (group A: 24.7%, group B: 26.6%). The CH content in RLP after treatment with bezafibrate showed a significant decrease (67.9%). Apolipoprotein AI after treatment with bezafibrate showed a significant increase (10.9%). Apolipoprotein B100 after treatment with either drug decreased significantly (group A: 19.8%, group B: 23.4%). The urinary albumin excretion rate after treatment with either drug showed no significant change over 4 years. Bezafibrate and pravastatin appear to be useful in the preventive treatment of albuminuria as well as in lowering lipid levels in NIDDM patients. PMID- 11426589 TI - A novel zinc finger protein mRNA in human umbilical vein endothelial cells is profoundly induced by tumor necrosis factor alpha. AB - Zinc finger proteins are known to mediate various transcriptional control mechanisms and other cellular functions in human cells. Tumor necrosis factor alpha (TNFalpha) induces a variety of genes in human endothelial cells including A20, an antiapoptotic zinc finger protein. In order to identify other zinc finger protein genes induced by TNFalpha, we studied the gene expression profile of human umbilical vein endothelial cells (HUVECs) stimulated by TNFalpha by means of oligonucleotide microarrays. Among the 155 genes encoding zinc finger motif, the level of EST M88357 mRNA encoding a novel designated EZFIT (endothelial zinc finger protein induced by TNFalpha) was induced most profoundly (>19 fold). The EZFIT gene is located on the chromosome 19q13.4. Isolation of the full length cDNA coding sequence by PCR using primers architected from the genomic sequence revealed that EZFIT has 490 amino acids which contain 13 C2H2 zinc finger motifs. Among 24 human organs and cell types studied, EZFIT mRNA was found to be most highly expressed in the placenta followed by the brain, testis, pancreas, heart, small intestine, muscle, uterus, prostate and peripheral blood leukocytes. EZFIT mRNA was not detected in the liver, lung, colon, stomach, the salivary gland or the thyroid gland. PMID- 11426590 TI - Push-out testing and histological evaluation of glass reinforced hydroxyapatite composites implanted in the tibia of rabbits. AB - In vitro and in vivo bioactivity studies were performed to assess the biocompatibility of CaO-P2O5 glass-reinforced hydroxyapatite (GR-HA) composites. The ability to form an apatite layer by soaking in simulated body fluid (SBF) was examined and surfaces were characterized using FTIR reflection and thin-film X ray diffraction analyses. Qualitative histology, histomorphometric measurements, and push-out testing were performed in a rabbit model for characterizing bone/implant bonding. Under the in vitro conditions using SBF, an apatite layer could not be formed on GR-HA composites within 8 weeks. Results of push-out testing showed bonding between the composites and bone, ranging from 130-145 N after 2 weeks of implantation. After the longest implantation period, 16 weeks, the GR-HA composite prepared with the higher content of CaO-P2O5 glass showed the highest bonding force, 606 +/- 45 N, compared to 459 +/- 30 N for sintered HA. Development of immature bone and modifications in the turnover of a more mature bone on the surface of GR-HA composites were similar to those on sintered HA. PMID- 11426591 TI - Novel adhesion prevention membrane based on a bioresorbable copoly(ester-ether) comprised of poly-L-lactide and Pluronic: in vitro and in vivo evaluations. AB - Block copolymers consisting of poly(L-lactide) (PLLA) and poly(oxyethylene-co oxypropylene), with various compositions, were synthesized and characterized in vitro and in vivo for their application as postoperative adhesion prevention membranes. It was found that the flexibility and degradability of the cast films of the block copolymers grew with increasing Pluronic F68 [PN; poly(oxyethylene co-oxypropylene] composition. The receding contact angle of the copolymer films against water became lower than that of the PLLA film, because the surface was predominantly covered with more hydrophilic PN segments in a wet state. This surface property significantly affects the cell attachment property of the copolymer films, and the fibroblasts cultured on the films exhibit a spheroid like morphology. The copolymer films subcutaneously implanted in the back of rats induced milder tissue responses compared with PLLA homopolymers, because of the increased surface hydrophilicity in the former. In vivo evaluation using a uterus horn model in rats revealed that the performance of these copolymer films as an adhesion-prevention membrane is comparable to that of a conventionally utilized membrane of oxidized regenerated cellulose. These results indicate that the copolymer films are biocompatible materials with controllable mechanical properties and biodegradability as adhesion-prevention membranes. PMID- 11426592 TI - Physicochemical study of plasma-sprayed hydroxyapatite-coated implants in humans. AB - This study represents the first report of the physical and chemical changes occurring in coatings of failed hydroxyapatite (HA)-coated titanium implants obtained from a comprehensive, multicenter human dental implant study. A total of 53 retrieved samples were obtained and compared with unimplanted controls with the same manufacturer and similar manufacture dates. Forty-five retrieved implants were examined for surface characteristics and bulk composition. Implants were staged based on implantation history: stage 1 (implants retrieved between surgical placement and surgical uncovering), stage 2 (implants retrieved at surgical uncovering and evaluation), stage 3 (implants retrieved between surgical uncovering evaluation and occlusal loading), and stage 4 (implants retrieved after occlusal loading). Scanning electron microscopy showed progressive coating thinning with implantation time. At later stages, bare Ti metal was detected by energy-dispersive X-ray analysis and electron spectroscopy for chemical analysis. Increases in Ti and Al (2-7.5 atm % each) were detected at the apical ends of all stage 4 samples. In unimplanted coatings, X-ray diffraction analysis demonstrated the presence of amorphous calcium phosphate, beta-tricalcium phosphate, tetracalcium phosphate, and calcium oxide in addition to large hydroxyapatite crystals (c axis size, D002 = 429 +/- 13 A; a axis size, D300 = 402 +/- 11 A, a/c aspect ratio 0.92). The nonapatitic phases disappeared with increased implantation time, although there was a persistence of amorphous calcium phosphate. Bulk coating chemical analysis showed that Ca/P ratios for implant controls (1.81 +/- 0.01) were greater than stoichiometric HA (1.67) and decreased for implant stages 3 and 4 (1.69 +/- 0.09 and 1.67 +/- 0.09, respectively), explained by the dissolution of the non apatitic phases. Crystal sizes also changed with implantation times, being smaller than the control at all but stage 4. Fourier transform infrared analyses agreed with these results, and also indicated the accumulation of bone (protein and carbonate-apatite) in the retrieved coatings. The accumulation of bone was not stage dependent. These findings indicate that there was some biointegration with the surrounding bone, but the greatest changes occurred with the HA coating materials, their loss, and chemical change. PMID- 11426593 TI - A new bioactive bone cement: effect of glass bead filler content on mechanical and biological properties. AB - A new bioactive bone cement (designated GBC), consisting of bioactive glass beads as an inorganic filler and polymethylmethacrylate (PMMA) as an organic matrix, has been developed. The purpose of the present study was to examine the effect of the amount of glass bead filler added to GBC on its mechanical and biological properties, and to decide the most suitable content of filler. Serial changes in GBC with time were also examined. The newly designed bioactive beads, consisting of MgO-CaO-SiO2-P2O5-CaF2 glass, were added to the cement in the proportions 30, 40, 50, 60, and 70 wt %. These cements were designated GBC30, GBC40, GBC50, GBC60, and GBC70, respectively. The compressive strength and the elastic modulus of bending of GBC increased as the glass bead content increased. The various types of GBC were packed into the intramedullar canals of rat tibiae to evaluate osteoconductivity, as determined by an affinity index calculated as the length of bone in direct contact with the cement expressed as a percentage of the total length of the cement surface. Rats were killed at 4 and 8 weeks after the operation and the affinity index was calculated for each type of GBC. Histologically, new bone had formed along the surface of all types of GBC within 4 weeks, even in GBC30 containing only 30 wt % of glass beads. At each time interval studied, there was a trend for the affinity index of GBC to increase as the glass bead filler content increased. There was no significant increase of affinity index between GBC60 and GBC70. The affinity indices for all types of GBC increased significantly with time up to 8 weeks. The handling properties of GBC were comparable to those of conventional PMMA bone cement. We conclude that when mechanical properties and osteoconductivity are both taken into consideration, GBC60 is the most suitable formulation; it shows excellent osteoconductivity and sufficient mechanical strength for clinical use. PMID- 11426594 TI - Tissue responses around polymethylmethacrylate particles implanted into bone: analysis of expression of bone matrix protein mRNAs by in situ hybridization. AB - Tissue responses around implanted polymethylmethacrylate (PMMA) particles were analyzed by in situ hybridization with digoxigenin-labeled procollagen alpha1(I) (COL), osteonectin, osteocalcin, and osteopontin (OPN) mRNA probes. PMMA particles (150-300 microm in diameter) were implanted into rat tibiae, and specimens were collected at 3, 5, 7, and 10 days after operation. New bone was formed centripetally, and bone-forming osteoblasts expressed all four kinds of mRNAs. A COL signal was expressed most strongly and widely. In the early stage, COL-positive cells were detected on and among particles sporadically. A COL signal was rarely detected in cells on the surfaces of the particles, suggesting that PMMA particles may suppress osteoblast differentiation. Osteonectin and osteocalcin mRNAs were expressed in bone-forming osteoblasts in a similar pattern by day 7. By contrast, an OPN signal was detected mainly on the particles, not only in COL-positive osteoblasts but also in COL-negative round cells. The latter cells had acid phosphatase activity, suggesting that they might be macrophages responding to a foreign body. At day 10, an OPN signal was detected continuously in multinucleated cells on PMMA particles, whereas new bone was formed away from particles. Our approach helped us to understand the initial cellular reaction to materials, which may determine their biocompatibility. PMID- 11426595 TI - A novel surfactant-based immobilization method for varying substrate-bound fibronectin. AB - Most biomaterials can be rendered adhesive for anchorage-dependent cells by adsorption of serum, isolated extracellular matrix proteins, or immobilization of peptide sequences. However, difficulties are frequently encountered in characterizing the adsorbed layer due to conformational changes in the molecules following adsorption and interference from nonspecifically adsorbed molecules. In this study, we have investigated a technique for covalently immobilizing fibronectin to the PEO-containing triblock copolymer Pluronic F108 ("F108"). We have compared this technique to solution adsorption of fibronectin for its ability to provide controlled variation of bound fibronectin and regulation of fibroblast behavior. Both simple adsorption and covalent immobilization were effective for varying substrate-bound fibronectin. However, adsorption of fibronectin did not effectively regulate fibroblast attachment or spreading in either serum-free or serum-containing media. Fibroblast attachment, spreading, cytoskeletal organization, and proliferation were effectively regulated in response to fibronectin immobilized to F108. Furthermore, F108-treated surfaces without immobilized fibronectin did not support nonspecific fibroblast attachment, even in the presence of serum-containing medium. Fibroblasts were observed to only proliferate on surfaces with high levels of immobilized fibronectin that supported extensive cell spreading and cytoskeletal organization. In summary, covalent immobilization of fibronectin to F108 provided controlled regulation of fibroblast behavior without interference from nonspecific protein adsorption, even in the presence of serum-containing medium. PMID- 11426596 TI - Suppression of apoptosis in hepatocytes by fructose-modified dendrimers. AB - By immobilizing fructose-modified dendrimers on a polystyrene culture plate, the number of initially adhered hepatocytes on it was increased. Moreover, increasing the number of generations of fructose-modified dendrimer (fructose-dendrimer) increased the number. Urea synthesis per unit area also was increased, corresponding to the increase in the number of initially adhered hepatocytes. This result suggests that the fructose-dendrimers do not cause a decline in cell function. On the other hand, apoptosis of hepatocytes occurs during cultivation, and results in a decrease in the number of adhered cells and a decline in cell function. Fructose-dendrimers were found to suppress apoptosis of hepatocytes. This characteristic is considered to be responsible for the increase in the number of initially adhered hepatocytes without a decline in cell function. Fructose-dendrimers are shown to be very suitable scaffolds for use in a high performance bioartificial liver support system. PMID- 11426597 TI - Preparation and compressive strength of alpha-tricalcium phosphate/gelatin gel composite cement. AB - A novel alpha-tricalcium phosphate (TCP) and gelatin gel composite cement was prepared, and the effects of gelatin content, liquid/powder ratio, setting time, and additives (rod-like hydroxyapatite and CaTiO3 particles) on the microstructure and compressive strength of the setting product were investigated. Addition of gelatin gel to alpha-TCP cement resulted in the formation of a porous solid possessing pores of 20-100 microm in diameter whose pore diameter increased with increasing gelatin gel content. The compressive strength of alpha-TCP cement after 1 week increased from 9.0 to 14.1 MPa with increasing gelatin gel content up to 5 wt % and thereafter decreased. The compressive strength of the cement containing 5 wt % gelatin gel increased with time up to 35 MPa after 1 month whereas without gelatin gel it was approximately 20 MPa. Dispersion of 5 wt % of rod-like hydroxyapatite and CaTiO3 powders with alpha-TCP cement containing 5 wt % gelatin gel increased the compressive strength after 1 week from 14.1 to 31.3 and 34.8 MPa, respectively. PMID- 11426598 TI - Sensitizer contained in heat-decomposed dye. AB - Recently a new resorbable P(LA/CL) monofilament suture made of L-lactide and epsilon-caprolactone was developed. The sensitization test was carried out using a guinea pig maximization test according to the Japanese Guidelines. As a result of this test, it was revealed that the methanol extract of P(LA/CL) suture contains a skin sensitizer. It was suggested that the D&C Violet No. 2 contained in P(LA/CL) sutures is decomposed during the melt-spinning process and that the decomposed dye induces skin sensitization. To characterize the sensitizer (the decomposed dye) it was separated and collected by HPLC. To determine the molecular formula of the sensitizer, MS, IR, and NMR spectra were obtained. The molecular weight of the sensitizer was estimated as 240 by measuring the MS. Considering the chemical structure of the dye and the molecular weights, it was determined that the sensitizer was 1,4-dihydroxyanthraquinone (quinizarin). The D&C Violet No. 2 contained in P(LA/CL) sutures was decomposed by the melt spinning procedure to form quinizarin, one type of anthraquinone dye, and the sensitizer was identified to be, indeed, quinizarin. To evaluate the sensitizing potential of quinizarin, we tested skin sensitization using the maximization test method. Quinizarin induced skin sensitization in guinea pigs, and the minimum sensitizing concentration was approximately 100 ppm. The concentration of quinizarin contained in the P(LA/CL) suture was 1-5 ppm, a value that is lower than the minimum sensitizing concentration of quinizarin. This suggests that P(LA/CL) suture has a low risk with regard to contact dermatitis. PMID- 11426599 TI - In vitro studies of platelet adhesion on laser-treated polyethylene terephthalate surface. AB - In order to improve blood compatibility, the surfaces of polyethylene terephthalate (PET) were treated using CO2 pulsed and KrF excimer lasers. The physico-chemical characterization of the laser-treated PET surfaces was carried out through attenuated total reflectance infrared spectroscopy and contact-angle measurements. The hemocompatibility of the laser-irradiated PET films was examined in vitro to evaluate their capability of inducing platelet adhesion in comparison with unmodified PET. The number of adhered platelets was determined by lactate dehydrogenase (LDH) activity measurement. Platelet adhesion on the untreated PET was relatively high compared to the laser-treated samples. Laser irradiation of PET surface reduced the number of adherent platelets and prevented platelet spreading on the surface. Reduction of platelet adhesion was attributed to the change in morphology, chemical structure, and crystallinity of the PET surface due to laser irradiation with various numbers of pulses. The morphology of adhered platelets on the PET surfaces was investigated by scanning electron microscopy (SEM). The SEM observations were consistent with the results obtained from LDH activity measurement. PMID- 11426600 TI - Correlations between osteocalcin content, degree of mineralization, and mechanical properties of C. carpio rib bone. AB - Osteocalcin is one of the most abundant noncollagenous proteins in bone. It is strongly associated with the mineral phase of bone, and has long been associated as a marker of bone turnover. However, its relationship to bone composition, strength, and structure is unclear. Carp rib bone is an excellent model for the study, because osteocalcin represents almost 60% of the total extractable noncollagenous proteins found in it. Because of the abundance of osteocalcin relative to other extractable proteins, any changes in the properties of carp rib bone would be more likely influenced by the osteocalcin concentration. To test the hypotheses that the concentration of osteocalcin is reflected in other properties of bone, the correlations between the osteocalcin concentration and the mineral content, microstructural properties, and physical characteristics of the bone mineral crystals were determined utilizing radioimmunoassay (RIA), spectrophotometry, nanoindentation, and small-angle X-ray scattering (SAXS) techniques, respectively. Osteocalcin concentration was found to be correlated to the molar Ca/P ratio and inversely correlated to the elastic modulus and hardness in the longitudinal plane. This study provides evidence for a putative relationship between the concentration of osteocalcin and the microstructural mechanical properties of bone. Correlations were also found between the mechanical properties in the longitudinal plane and both the phosphate content and the molar Ca/P ratio. However, no relationships could be identified between osteocalcin concentration and several parameters of bone crystals, as determined by SAXS. PMID- 11426601 TI - Nanoindentation study of interfaces between calcium phosphate and bone in an animal spinal fusion model. AB - Intertransverse process spinal fusion is a common surgical procedure for the treatment of spinal disorders. In the present study, a porous hydroxyapatite (HA)/beta-tricalcium phosphate (beta-TCP) ceramic was tested as graft material using a rabbit lumbar transverse process (L5-L6) fusion model. The porous ceramic blocks were implanted onto the dorsal decorticated surface of the lumbar transverse processes. The specimens were harvested at the seventh week after implantation. Histomorphological observation revealed that the integration of HA/beta-TCP with the host bone of the transverse process occurred by both cancellous bone formation and cartilage formation. Scanning electron microscopy wavelength dispersive X-ray spectrometry examinations showed significant differences in calcium, phosphorus, and sulfur contents in the newly formed tissues and the porous HA/TCP implants. Nanoindentations were used to evaluate the intrinsic mechanical properties of the implants and the newly formed tissues. The Young's moduli of the newly formed cartilage, new cancellous bone, and HA/TCP, were 0.66 +/- 0.02 GPa, 2.36 +/- 0.50 GPa, and 10.2 +/- 1.21 GPa, respectively. Nanoindentation results revealed degradation of the porous ceramics and incomplete calcification of the new cancellous bone at the seventh week after implantation. Nanoindentation appeared to be a useful technique for assessing the mechanical status of spinal fusion in animal models. PMID- 11426602 TI - Long-term in vivo bioactivity and degradability of bulk sol-gel bioactive glasses. AB - Melt-derived bioactive glasses have been used with success in various clinical applications for over 10 years. Recently, particles of sol-gel-derived bioactive glasses with an initial high specific surface area have been shown to exhibit excellent osteoconductive properties as well as significant degradability. In this work, we explored the long-term in vivo bioactivity and degradability of bulk sol-gel-derived glasses in a rabbit model. Two sol-gel compositions (58S and 77S Bioglass) were used. Bulk 45S5 Bioglass was used as a control. Both sol-gel derived glasses demonstrated osteoconductive properties similar to 45S5 Bioglass. In addition, absorbability was observed for both sol-gel-derived glasses starting after 12 weeks of implantation. Total absorption reached 40% after 52 weeks. No degradation could be measured in the case of bulk 45S5 melt-derived Bioglass within 1 year of implantation. The degradation process was highly time dependent, as demonstrated by regression analysis. New bone formation was found to fill in areas that had been resorbed, similar to bone remodeling. This absorbability can be assumed to be at least partially related to an osteoclastic resorption as viable osteoclasts-like cells were found to be in direct contact with the glass surfaces. PMID- 11426603 TI - Polishing and coating carbon fiber-reinforced carbon composites with a carbon titanium layer enhances adhesion and growth of osteoblast-like MG63 cells and vascular smooth muscle cells in vitro. AB - Carbon fiber-reinforced carbon composites (CFRC) are considered to be promising materials for orthopedic and dental surgery. Their mechanical properties can be tailored to be similar to those of bone, and their chemical composition (close to pure carbon) promises that they will be tolerated well by the surrounding tissue. In this study, CFRC composites were fabricated from phenolic resin and unidirectionally oriented Torayca carbon fibers by carbonization (1000 degrees C) and graphitization (2500 degrees C). The material then was cut with a diamond saw into sheets of 8 x 10 x 3 mm, and the upper surface was polished by colloidal SiO2 and/or covered with a carbon-titanium (C:Ti) layer (3.3 microm) using the plasma-enhanced physical vapor deposition method. Three different kinds of modified samples were prepared: polished only, covered only, and polished + covered. Untreated samples served as a control. The surface roughness of these samples, measured by a Talysurf profilometer, decreased significantly after polishing but usually did not decrease after coating with a C:Ti layer. On all three modified surfaces, human osteoblast-like cells of the MG63 line and rat vascular smooth muscle cells (both cultured in a Dulbecco's minimum essential medium with 10% fetal bovine serum) adhered at higher numbers (by 21-87% on day 1 after seeding) and exhibited a shorter population doubling time (by 13-40%). On day 4 after seeding, these cells attained higher population densities (by 61 378%), volume (by 18-37%), and protein content (by 16-120%). These results were more pronounced in VSMC than in MG63 cells and in both groups of C:Ti-covered samples than in the polished only samples. The release of carbon particles from the CFRC composites was significantly decreased--by 8 times in the polished only, 24 times in the covered only, and 42 times in the polished + covered samples. These results show that both polishing and carbon-titanium covering significantly improve the biocompatibility of CFRC composites in vitro, especially when these two modifications are combined. PMID- 11426604 TI - In vitro bioactivity and structural features of mildly heat-treated sol-gel derived silica fibers. AB - The ability of sol-gel-derived silica fibers heat treated at a low temperature to induce formation of bone-like calcium phosphate (HCA) on their surfaces provides alternatives for the design of novel biomaterials, for example as implants used in tissue guiding or bone repairs. In this study, dry spinning was used to prepare the sol-gel fibers, which were heat-treated at 175 degrees and 250 degrees C. In addition, the differences in the surface topography (in a nanometer scale) of different fibers with respect to their in vitro bioactivity were studied. The structure of the fibers was varied using three different factors: (1) spinnable sols having varying structures and sizes of silica polymers to establish varying viscosity levels; (2) aging of green-state fibers; and (3) heat treatment of fibers. The in vitro bioactivity and solubility tests were done in simulated body fluid (SBF). To monitor surface topography and roughness of the heat-treated silica fibers, a scanning probe microscopy (SPM) with tapping mode AFM was used. Different fibers obtained clearly different properties. The fibers spun at about eta > 3.0 Pas had the best properties with respect to bioactivity, especially when they were heat-treated at 175 degrees C. It was found that surface structure in a nanometer scale was the most important factor controlling the in vitro bioactivity of heat-treated silica fibers. The correct proportions between the peaks and peak distances at the surfaces are suggested to be important with respect to in vitro bioactivity. The results indicate that peak distance distribution between 5-50 nm, especially between 5-20 nm, together with a peak height > or = 1 nm is most favorable for calcium phosphate formation. PMID- 11426605 TI - Observations at the articular surface of hip prostheses: an analytical electron microscopy study on wear and corrosion. AB - We used scanning electron microscopy in combination with X-ray microanalysis to evaluate Co-, Cr-, and Mo-based human femoral hip prostheses. In total, 23 retrieved implants and four new implants were included in this study. Scanning electron microscopy of the polished surface of all arthroplasties showed, in addition to the polishing marks, small round and angular holes or pits. Other types of surface irregularities were interpreted as wear or corrosion of the metal compound. In all cases studied, corrosion propagated from holes at the surface of the polished prosthesis heads, in some cases also along phase boundaries. X-ray microanalysis of the intact prosthetic surface showed a relative composition of the elements Co, Cr, and Mo, which was in agreement with the manufacturer's information (63:33:4%). However, X-ray microanalysis spot analysis of the surface holes showed deviation in the relative composition of the elements Co, Cr, and Mo and also the presence of Si, Ti, and Al. Furthermore, Ti and Al could be traced back at an artificially made fracture plane of a new prosthesis. Therefore, Ti and Al have to be present during the manufacturing process. These impurities in the metal prosthesis alloy may create a galvanic element with the Co, Cr, Mo alloy of the implant. If this is the case, such a galvanic element in combination with the electrolyte environment formed by body fluids, can induce galvanic corrosion with release of metal particles. PMID- 11426606 TI - Evaluation of plasma polymer-coated contact lenses by electrochemical impedance spectroscopy. AB - The influence of ultra-thin (i.e., 5-50 nm) plasma polymer coatings on siloxane based hydrogel contact lenses was investigated by electrochemical impedance spectroscopy (EIS). Impedance measurements as a function of frequency (Bode plots) were taken at regular intervals until steady impedance was obtained, indicating that the lenses were saturated. Appropriate equivalent circuit models were constructed to describe the salt intrusion characteristics of the plasma polymer-coated contact lenses. This provided information pertaining to the resistance and capacitance of interfacial and bulk layers in the plasma polymer coated lenses. Resistance relates to ion permeability and capacitance for water uptake. This investigation showed that some of an ultra-thin layer of plasma polymer applied onto a dry hydrogel remains a contiguous film after the substrate hydrogel swells upon hydration. In some cases, however, the overall impedance of the coated lens is so low that the state of the plasma polymer layer after the hydration of the substrate hydrogel could not be judged by EIS. PMID- 11426607 TI - Low-temperature synthesis, structure, and bioactivity of gel-derived glasses in the binary CaO-SiO2 system. AB - Glasses in the binary system CaO-SiO2 for which the molar fraction of CaO is 0 < or = x < or = 0.50, were prepared by means of a sol-gel route starting from tetraethylorthosilicate and calcium nitrate. The textural features of the monoliths obtained were characterized using N2 gas adsorption, helium ultrapycnometry, and mercury porosimetry. In vitro bioactivity tests were performed in simulated body fluid (SBF). The ionic concentration of the SBF after glass immersion was analyzed using inductively coupled plasma atomic emission spectroscopy. The surfaces of the specimens were characterized using X-ray diffraction and scanning electron microscopy coupled with energy dispersive X-ray analysis before and after in vitro testing. The textural characterization revealed that the glasses were mesoporous with cylindrical pores with average diameters ranging from 25 to 663 A depending on the molar fraction of CaO. The in vitro studies showed that all binary CaO-SiO2 gel-glass compositions produced were bioactive. These results indicate that the binary gel-derived CaO-SiO2 system exhibits a level of bioactivity over a similar molar range of SiO2 content as the previously studied ternary CaO-P2O5-SiO2 system. PMID- 11426608 TI - Bone bonding in sintered hydroxyapatite combined with a new synthesized agent, TAK-778. AB - We studied the stimulatory effects of TAK-778, a new synthetic 3-benzothiepin derivative that promotes osteoblast differentiation, in bone bonding to sintered hydroxyapatite implants in rabbit tibiae. Smooth-surfaced rectangular plates (15 x 10 x 2 mm) made of sintered hydroxyapatite were implanted into the proximal metaphyses of bilateral rabbit tibiae, with TAK-778-containing sustained-release microcapsules packed into the medullary cavity in one limb and untreated microcapsules packed in the contralateral limb to serve as a paired control. At 4, 8, and 16 weeks after implantation, bone bonding at the bone-implant interfaces was evaluated by a detaching test and undecalcified histological examination. The tensile failure load increased from 4 to 16 weeks for both groups; however, the tensile failure load of the TAK-778-treated group was significantly greater than that of the control group at each interval after implantation. Histologically, the TAK-778-treated specimens showed greater active new bone formation mainly in the medullary cavity and more extensive bonding between the implant and bone than the untreated specimens. The results of this study suggest that adding osteoinductive TAK-778 to hydroxyapatite implants may significantly accelerate bone apposition to the implants and improve the bonding process at the interface. This would help to establish an earlier and stronger bonding of orthopedic ceramic implants between the surrounding bone tissue. PMID- 11426609 TI - Excessive apoptosis in low risk myelodysplastic syndromes (MDS). AB - The paradox of peripheral cytopenias despite a normo/hypercellular marrow in MDS has been ascribed to excessive intramedullary hematopoietic cell apoptosis. Programmed cell death (PCD) in early disease might be triggered by the BM microenvironment, mediated either through inhibitory cytokines such as tumor necrosis factor alpha (TNF-alpha) or fas/fas ligand signaling or through a relative deficiency in hematopoietic growth factors. Intrinsic cellular defects giving rise to abnormalities in cell-cell or cell-stromal interaction, cell signaling or cell cycling may also underlie hematopoietic progenitor apoptosis. Alternatively, an early 'hit' in the multistep pathogenesis of MDS may result in a higher proliferative rate of the neoplastic clone. Increased apoptosis may thus represent a homeostatic process to control cell numbers. This paper shall summarize current evidence implicating a role for increased PCD in low risk MDS, outline possible etiologic factors and suggest potential therapeutic mechanisms whereby excessive hematopoietic progenitor cell apoptosis might be circumvented. PMID- 11426610 TI - Vasculitis and leukemia. AB - Vasculitis may accompany neoplasias and be of paraneoplastic type or associated with drugs used in patient treatment. We evaluated skin biopsies of twenty-eight cases with vasculitis accompanying leukemias reviewed and clinical outcome was evaluated. Eleven of the 28 cases had paraneoplastic vasculitis and 17 had vasculitis associated with various drugs including chemotherapy, cytokines and antibacterial agents. Paraneoplastic vasculitis was seen in 3 cases with chronic myelocytic leukemia in blastic phase, 5 patients with acute myeloblastic leukemia, and 3 with myelodysplastic syndrome. Drugs responsible for the 17 cases of vasculitis included hydroxyurea, vincristine, cytosine-arabinoside, methotrexate, all-trans retinoic acid, granulocyte-colony stimulating factor, interferon and antibiotics. Paraneoplastic vasculitis is not rare in leukemias and may be a manifestation of the blastic phase of chronic myeloid leukemia. Furthermore paraneoplastic vasculitis can be fatal in myelodysplastic syndromes and may be present clinically before the specific diagnosis is made. Drugs used in routine therapy may be the cause of the vasculitis, thus skin biopsy should be performed in all cutaneous lesions in patients with hemopoietic neoplasias. PMID- 11426611 TI - Lack of efficacy of 2-chlorodeoxyadenoside in the treatment of splenic lymphoma with villous lymphocytes. AB - Splenic lymphoma with villous lymphocytes (SLVL) is a B-cell chronic lymphoproliferative disorder. Splenectomy and/or chlorambucil (CLB) are usually regarded as the most effective treatment in SLVL patients. However, a few patients relapse and the second line therapy remains questionable. Although 2-Cda has been evaluated in patients with chronic lymphoid leukemia (CLL) and hairy cell leukemia (HCL), it has been reported as the treatment of SLVL in only one case report. Therefore, we have evaluated its efficacy and toxicity in 7 SLVL patients. The median duration between diagnosis and treatment was 18 months (range, 1 to 59). The patients received 2-CdA (0.1 mg/kg/d) by venous infusion for 7 days with a median number of 1 cycle (range, 1 to 2) either as a first line therapy (one patient) or after a failure of other therapies (splenectomy, chemotherapy). Two patients achieved a complete response. The first one maintained his CR during a follow-up of 9 months and then relapsed; the second patient remained in CR after a follow-up of 20 months. Four patients achieved a partial response and relapsed after a median follow-up of 3.5 months (range, 1 to 4). One patient had no response. The treatment was not well tolerated with many infectious events. In the limits of our study, 2-Cda does not appear to be efficient therapy for SLVL and is not well tolerated for patients in relapse after splenectomy or resistant to CLB. PMID- 11426612 TI - The significance of autoantibodies in non-Hodgkin's lymphoma. AB - Lymphoproliferative diseases are the most common disorders associated with autoimmune disturbances. We determined the autoimmune phenotype of 64 non Hodgkin's lymphoma patients' and compared their clinicopathologic properties. Serum direct antiglobulin test [(DAT) n=64], indirect antiglobulin test [(IAT) n=61], platelet autoantibodies [(PAA) n=51], anti nuclear antibodies [n=33], anti native DNA [n=29], anti phospholipid antibodies [n=40] and, lupus anticoagulant [n=33] were used as autoimmune markers. Twenty five patients (39%) displayed one or more autoimmune marker positivity (+). Three patients with (+) DAT and IAT had autoimmune hemolytic anemia and two patients with PAA had autoimmune thrombocytopenia. Male patients were more susceptible to autoimmunity in low grade lymphomas and the statistical difference was significant (p=0.035). Most of the autoimmune markers (+) patients had low grade and disseminated disease but this was not significant. Remission rates were not found to be different between autoimmune marker (+) and (-) patients. Although statistically not significant. median survival was longer in autoimmune marker (-) patients than in the others (50 versus 39 months). The significance of autoimmunity in NHL in a larger series of patients should be investigated in future studies. PMID- 11426613 TI - Primary non-Hodgkin's lymphoma of the gall bladder. AB - Primary non-Hodgkin lymphoma of the gallbladder is a very rare location of extranodal non-Hodgkin lymphomas. A patient with a primary non-Hodgkin lymphoma of the gallbladder is reported and in addition, the English literature is reviewed. Clinical presentation, diagnostic evaluation, histopathologic findings, treatment modalities and prognosis of primary gallbladder lymphomas reported up to date are reviewed and discussed. Our patient was diagnosed as a T-cell lymphoblastic lymphoma, after cholecystectomy, and had no evidence of disease elsewhere. She was treated with combination chemotherapy and complete remission was achieved. She remains free of disease 9 years later. Review of the literature over a 30-year period revealed only 12 cases of well-documented primary non Hodgkin lymphoma involvement of the gallbladder, including the present case. Patients present clinically with symptoms and signs indicating either biliary tract pathology or a gastrointestinal tumor. Diagnostic investigation included ultrasound of the upper abdomen, computed tomography of the abdomen and pelvis, oral cholecystography, percutaneous cholangiography and endoscopic retrograde cholangiopangreatography. Preoperative diagnosis was established in none of the patients. Treatment modalities included surgery and postoperative chemotherapy and irradiation. The prognosis is overall poor and only 2 patients are alive after 1 and 9 years respectively, the latter being our case. Here we document the first reported case of a patient with primary T-cell lymphoblastic non-Hodgkin lymphoma of the gallbladder. Review of the literature shows the existence of non Hodgkin lymphoma of the gallbladder, its rarity and its general dismal prognosis. PMID- 11426614 TI - Salvage therapy for relapsed mediastinal B-cell lymphoma with allogeneic HLA identical related donor bone marrow transplantation, donor lymphocyte infusion and IDEC-C2B8. AB - Primary B-cell lymphoma of the mediastinum is an aggressive non-Hodgkin's lymphoma with distinct clinicopathologic features. Response rates are between 60 80% following intensive chemotherapy regimens. Poor responders or patients with an early relapse usually do not achieve a prolonged second remission with conventional salvage therapy protocols and therefore qualify for intensive or experimental approaches. Here we describe two patients of same age, gender and stage with primary mediastinal B-cell lymphoma and an early relapse after the first courses of combination chemotherapy and irradiation of the mediastinum. One patient relapsed after a salvage therapy with allogeneic donor-related bone marrow transplantation and donor lymphocyte infusion but responded again with a continuing good partial remission after infusion of the chimeric anti-CD20 antibody IDEC-C2B8. For the other patient an allogeneic bone marrow transplantation was not possible. He finally failed to respond to salvage therapy with IDEC-C2B8 and died of progressive disease. The anti-CD20 antibody IDEC-C2B8 induced a partial remission in a patient with primary mediastinal B-cell lymphoma refractory to other therapeutic approaches, including allogeneic bone marrow transplanatation (alloBMT), donor lymphocyte infusion (DLI) and irradiation. The role of IDEC-C2B8 as a component of salvage regimens appears to be worthy for further evaluation in high-risk patients with primary mediastinal B-cell lymphoma PMID- 11426615 TI - Correction of disease related anaemia of B-chronic lymphoproliferative disorders by recombinant human erythropoietin: maintenance is necessary to sustain response. AB - Thirty three B-chronic lymphoproliferative disorder (B-CLD) patients [22 with B chronic lymphocytic leukemia (B-CLL), 5 with small lymphocytic lymphoma (SLL) and 6 with lymphoplasmacytic lymphoma (LPL)] with anaemia (Ht <32%) of no other cause but their disease, received recombinant human erythropoietin (r-HuEPO). The treatment protocol provided r-HuEPO in a dose of 150 U/kg s.c. thrice weekly for 3 mo. After 1.5 mo of r-HuEPO administration, if response was not satisfactory, r HuEPO dose escalation was utilised by giving incremental doses of 50 U/kg more than the previous dose up to a maximum dose of 300 U/kg tiw. After maximal response, half of the responding patients discontinued therapy, while the other half received maintenance therapy at a dose of 150 U/kg s.c./w. Oral iron was given throughout the study. Pretreatment EPO levels were determined in all patients. A complete response (CR) was defined when Ht was >38% and a partial response (PR) when there was an increase of the Ht >6% from the initial value was achieved. Sixteen of the 22 B-CLL patients had Rai stage III disease and 6 stage IV, with a median duration of anaemia 27 months (6-38); twelve of them were receiving chlorambucil while the rest were on no treatment. Of the SLL and LPL group, 4 patients had Ann Arbor stage III disease and 7 stage IV with a median duration of anaemia 24 months (5-36); 8 patients were on chlorambucil. Complete response was achieved in 50% of the B-CLL group and 54% of the SLL and LPL group, with an overall response rate of 77% and 81% respectively. All patients on maintenance therapy had a continuous response, while all patients, in whom rHuEPO was discontinued, relapsed. No correlation was found between patients: with low or high pretreatment serum EPO levels; those receiving concomitant therapy or not; those with B-symptoms or not; those with a non-diffuse or diffuse bone marrow infiltration pattern; and with splenomegaly or not. Life quality was significantly improved and no major side effects were encountered. We conclude from our study that r-HuEPO is very effective in correcting disease-related anaemia in B-CLD, resulting in down-staging of Rai stage III patients and that maintenance therapy is necessary. Whether the correction of anaemia improves patients' overall survival, still remains to be seen. PMID- 11426616 TI - Stimulation of Bruton's tyrosine kinase (BTK) and inositol 1,4,5-trisphosphate production in leukemia and lymphoma cells exposed to low energy electromagnetic fields. AB - We examined the effects of low energy electromagnetic field (EMF) exposure on the BTK kinase activity in B18-2 ([Btk-, rBTK(wt)] DT40) chicken lymphoma B cells and NALM-6 leukemic pre-B cells. Exposure of B 18-2 cells to EMF resulted in activation of BTK within 1 to 15 minutes in 8 of 8 independent experiments with stimulation indexes ranging from 1.2 to 13.3. While in some experiments the BTK stimulation was transient, in others the BTK activity continued to be significantly elevated for up to 4 hours. Similarly, exposure of NALM-6 cells to EMF resulted in activation of BTK within 30 minutes in 7 of 7 experiments with stimulation indexes ranging from 1.2 to 7.4. Stimulation of BTK activity in EMF exposed cells was associated with enhanced phosphoinositide turnover and increased inositol-1,4,5-trisphosphate (IP3) production in 7 of 13 experiments with DT40 cells and 7 of 13 experiments with NALM-6 cells. The likelihood and magnitude of an IP3 response after EMF exposure were similar to those after BCR ligation on DT40 cells and CD19 ligation on NALM-6 cells. These results confirm and extend our previous studies regarding EMF-induced biochemical signaling events in B-lineage lymphoid cells. PMID- 11426617 TI - Vascular density (tumor angiogenesis) in non-Hodgkin's lymphomas and florid follicular hyperplasia: a morphometric study. AB - Tumor angiogenesis is required for tumor growth and metastasis. A statistically significant correlation has been demonstrated between prognosis and the microvessel density (a measure of tumor angiogenesis) of solid tumors, particularly of the breast and prostate, and lymphoid neoplasms. The aim of this study was to establish whether a correlation exists between vascular density and the malignant category of Non-Hodgkin's Lymphoma (NHL) defined by two classification systems (Kiel and Working Formulation). We also tested whether florid follicular hyperplasia (FFH) and follicular lymphomas (FL) behave as new vessel stimulating conditions. Eighty-nine NHL lymph node biopsies were reviewed and categorized according to the Kiel Classification and Working Formulation. Twelve FL were also selected and compared to 12 FFH biopsies. Vessels were highlighted by immunostaining with anti-Factor VIII antibody and quantified both by counting higher vascular density fields and by estimating the proportional vascular area. The results showed a statistically significant difference between low and high grade NHL, when classified in either the Working Formulation (p=0.0015) or the Kiel Classification (p=0.002). No differences were found in vessel counts between Working formulation intermediate and high grade lymphomas. Vascular density is similar when FFH and FL interfollicular areas are compared. PMID- 11426618 TI - Lovastatin induces a pronounced differentiation response in acute myeloid leukemias. AB - We recently identified HMG-CoA reductase, the rate-limiting enzyme of the mevalonate pathway, as a potential therapeutic target of various retinoic acid responsive cancers. Lovastatin, a competitive inhibitor of HMG-CoA reductase, induced a retinoic acid-like differentiation response followed by extensive apoptosis in neuroblastoma cell lines at relatively low concentrations (<20 microM) of this agent. More recently, we demonstrated that acute myeloid leukemias but not acute lymphocytic leukemias also displayed increased sensitivity to lovastatin-induced apoptosis. In this study, we examined the ability of lovastatin to induce differentiation of acute myeloid leukemic cells and to evaluate the role differentiation may hold in the anti-leukemic properties of this agent. Increased expression of the leukocyte integrins CD11b and CD18 as well as down-regulation of the anti-apoptotic gene bcl-2 are associated with late stage differentiation of the myeloid lineage and retinoic acid induced maturation of acute myeloid leukemic cells. Lovastatin exposure induced increased expression of CD11b and CD18 markers similar to retinoic acid treatment. Following 24 hrs exposure to 20 microM lovastatin, all 7 acute myeloid leukemia cell lines tested showed a decrease in bcl-2 mRNA expression while only 1/5 acute lymphocytic leukemia cell lines showed a similar response. A role for bcl-2 in the apoptotic response of acute myeloid leukemia cells to lovastatin was demonstrated as exogenous constitutive expression of bcl-2 in the AML-5 cell line inhibited apoptosis in a time and dose dependent manner. Thus, lovastatin exposure of acute myeloid leukemia cells induced a differentiation response that may contribute to the therapeutic potential of this agent in the treatment of this disease. PMID- 11426619 TI - Thrombopoietin, interleukin-11, and early-acting megakaryocyte growth factors in human myeloid leukemia cells. AB - In this study we report our data on effects of early-acting megakaryocyte growth factors, particularly the c-mpl ligand also known as thrombopoietin (TPO) and interleukin-11 (IL-11), on cell proliferation and apoptosis (Apo) of primary acute myeloid leukemia (AML) cells. A proliferative response to TPO was noticed in the majority of AML samples (17/19) with an average increase of S-phase cells from 7.8% +/- 1.5 to 14.5% +/- 2.1 (p=0.0006). Resulting cell cycle activation did not always correlate with expression of the c-mpl receptor, although it was coupled, in the majority of samples, by an average decrease of apoptotic cells from 13% +/- 0.7 to 8.8% +/- 1.8 (p=0.05). Clonogenic cell growth (CFU-L) was confirmed in 5/17 of the samples with a mean colony number of 21.4 +/- 9.6 x 10(5) cells plated. Conversely, effects of IL-11 on AML cells demonstrated that cell cycle changes (recruitment from G0 to S phase) were promoted only in a minority of samples (2/14) and there was little, if any, effect on CFU-L growth (mean colony number=17.5 +/- 9.5) or Apo (from 13% +/- 0.7 to 13.3 +/- 1.9). Combination of TPO with IL-11 induced a slight increase of clonogenic cell growth, while the addition of IL-3 or SCF to the c-mpl ligand significantly raised the mean colony numbers up to 119.2 +/- 68.3 and 52.9 +/- 22.1 x 10(5) cells plated, respectively. In summary, TPO shows activity on AML cells by stimulating their proliferation in a significant proportion of cases and generally protecting the majority of AML blast cells from induction of Apo. Conversely, IL-11 exerts little effect on the cell cycle activation and Apo. These data help to understand regulation of myeloid leukemia cell growth and should be considered in the clinical use of early-acting megakaryocyte growth factors in acute leukemia. PMID- 11426620 TI - Differential effects of the MDR1 (multidrug resistance) gene-activating agents on protein kinase C: evidence for redundancy of mechanisms of acquired MDR in leukemia cells. AB - Human leukemia cells may acquire MDR1/P-glycoprotein-mediated multidrug resistance (MDR) in the course of short-term (within hours) exposure to many stress stimuli. This effect is thought to be associated with the activity of protein kinase C (PKC) (Chaudhary, Roninson, 1992. 1993). However, we show here that cytosine beta-D-arabinofuranoside (Ara C) and 12-O-tetradecanoylphorbol 13 acetate (TPA), agents that activated the MDR1 gene in the H9 T-cell leukemia line, caused different effects on PKC. Namely, TPA activated PKC whereas Ara C was without the effect. Furthermore, cell permeable ceramide, a lipid messenger known to mediate cellular effects of chemotherapeutic drugs and TPA, activated the MDR1 gene and down-regulated PKC. These results suggest that the MDR1 gene can be activated via the pathway(s) that requires PKC activity as well as via bypass of PKC. The redundancy of signaling pathways that regulate the acquisition of MDR should be taken into consideration for prevention of secondary drug resistance in hematological malignancies. PMID- 11426621 TI - Occurrence of angioimmunoblastic T cell lymphoma in a patient with chronic myelomonocytic leukemia features. AB - In a patient with recently diagnosed chronic myelomonocytic leukemia features, the biopsy of a peripheral lymphadenopathy seven months later revealed disorganised lymphoid tissue with a few large EBER (+) LMP1 (+) B-lymphocytes before any treatment was given. At this time, a clonal TCR gamma rearrangement and very faint clonal IgH rearrangement were demonstrated, and the diagnosis of angioimmunoblastic T-cell lymphoma was made. Treatment with MOPP was started, followed by Hydroxycarbamide and CHOP but the outcome was fatal. During the evolution, there was no blastic transformation of the chronic myelomonocytic leukemia. The T-cell lymphoma extended to abdominal lymph nodes, Waldeyer ring and bone marrow and the percentage of large LMPI EBER (+) B-cells increased in the lymph nodes. These findings do not support a common stem cell abnormality leading to myelodysplasia in the bone marrow and lymphoma in peripheral lymph nodes. The lack of a clearcut light chain restriction in the EBV infected B-cell is suggestive of a persistant EBV infection in polyclonal or oligoclonal activated B-cells as described in immunodepressed patients. The association of CMML features and an angioimmunoblastic T-cell lymphoma is discussed. PMID- 11426622 TI - Peroneal mononeuropathy in pediatric Hodgkin's disease. AB - A 12-year-old boy with Hodgkin's disease developed left peroneal nerve palsy during combination therapy with chemotherapy and low-dose irradiation. The palsy occurred twice; around 1-2 weeks after the second administration of vincristine in the second and third COPP (cyclophosphamide, vincristine, prednisolone, and procarbazine) regimens. Without any treatment, the peroneal neuropathy completely resolved clinically three months and electromyographically six months after the onset. He used to play television games for more than 6 hours a day with the legs crossed while sitting on the bedside. Compared to adult patients, little is known about the relationship between peroneal neuropathy and systemic malignant diseases in pediatric patients. This case shows for the first time that habitual leg crossing during potentially neurotoxic chemotherapy could induce peroneal mononeuropathy in a pediatric cancer patient. PMID- 11426623 TI - Plasma cell dyscrasia with polyneuropathy--POEMS syndrome presenting with vasculitic skin lesions and responding to combination chemotherapy. AB - We report a 61-year-old male patient who presented with severe sensorimotor neuropathy, leg edema and skin lesions with M-paraprotein and 50% plasma cells in the bone marrow. The POEMS (Crow-Fukase) syndrome was diagnosed and the skin lesions were compatible with vasculitis according to the histopathology. The patient was treated with aggressive combined chemotherapy, which induced improvement in both the clinical and laboratory parameters of his disease. To the best of our knowledge this is the first report of a vasculitic process underlying the skin changes in the POEMS syndrome. Our findings may shed light on the unknown pathogenesis of this syndrome and the successful results of treatment support the adoption of an aggressive therapeutic approach in symptomatic patients. PMID- 11426624 TI - Relapse of chronic myelogenous leukemia 12 years after allogeneic marrow transplantation: successful second transplantation with allogeneic peripheral blood progenitor cells. AB - We report on a 31-year old female patient who relapsed with CML in blast crisis 12 years after a successful BMT for CML in chronic phase from her HLA-identical sister. Because of her good performance status and the long time elapsed since her first BMT, PBPC transplantation of the originial donor was planned. Therefore, the patient was conditioned with busulfan and cyclophosphamide and then received unmanipulated PBPCs from her sister. GVHD prophylaxis consisted of MTX and CsA. She had early engraftment but considerable hepatotoxicity which resolved after more than six months. Furthermore, she developed acute GVHD of the skin grade 2, which responded to corticosteroids. Fifteen months after second transplantation the patient is alive and well in molecular remission and without signs of chronic GVHD. PMID- 11426625 TI - A case of meningeal myeloma presenting as obstructive hydrocephalus--a therapeutic challenge. AB - Meningeal involvement in myeloma is a rare though recognised complication, and previous treatment strategies have included radiotherapy, intra-thecal and systemic chemotherapy to which there is almost universally a poor response. We report a case of a 60-year-old lady in serological remission with IgG lambda myeloma, presenting with obstructive hydrocephalus due to meningeal infiltration treated with neurosurgical decompression. Such presentation and the treatment strategies are previously unreported, though in view of the ultimate resistance to treatment the prognosis remains poor. PMID- 11426626 TI - Homing-associated cell adhesion molecule (H-CAM/CD44) on human CD34+ hematopoietic progenitor cells. AB - Human CD34+ hematopoietic progenitor cells (HPCs) express CD44 and can directly adhere to hyaluronate (HA) via CD44. Furthermore, CD44 may also be involved in the regulation of CD34+ HPC proliferation and development. The expression of CD44 molecules on CD34+ hematopoietic progenitor cells is significantly lower on bone marrow (BM) CD34+ cells compared with circulating CD34+ cells in cord blood and peripheral blood. Myeloid and erythroid progenitor cells are found predominantly in CD34+ CD44+ cell fractions. More interestingly, CD34+ CD44- cells expressing B lymphocyte-associated CD10 and CD19 would represent unique B-lymphocyte committed precursors in the BM, which might undergo apoptotic cell death in the early steps of B-cell differentiation. PMID- 11426627 TI - The TEL-AML1 fusion accompanied by loss of the untranslocated TEL allele in B precursor acute lymphoblastic leukaemia of childhood. AB - The TEL-AML1 fusion which results from the t(12;21) rearrangement in childhood B precursor acute lymphoblastic leukaemia (B-precursor ALL) is often accompanied by loss of the untranslocated TEL allele. From 32/109 children with B-precursor ALL screened for these abnormalities, we found evidence for del 12p, including the loss of the untranslocated TEL allele, to be the secondary event to take place in the leukaemic cells from those patients positive for these abnormalities. This suggests that the initial or predisposing event is the generation of a TEL-AML1 fusion, followed by the promoting event of a deletion of a gene(s) on 12p. A striking characteristic of the leukaemic cells in 61% of the patients showing t(12:21). was the substantial evolution of the primary clonal line containing the reciprocal TEL-AML1 fusion. We were able to show loss of normal TEL in the same patients by interphase fluorescence in situ hybridisation (FISH) analysis and reverse-transcriptase polymerase chain reaction (RT-PCR). PMID- 11426628 TI - Telomere dynamics and genetic instability in disease progression of chronic myeloid leukemia. AB - Chronic myeloid leukemia (CML) is characterized by a Philadelphia (Ph) translocation creating a novel BCR-ABL oncoprotein, and CML patients have a chronic phase for several years followed by an intractable blast cell proliferation, called blast transformation. In the blast phase, more than 60% of patients show additional cytogenetic changes, e. g., double Ph, +8, i(17q). In this review, we would like to address genetic changes, including genome instability, cytogenetic changes, and telomere dynamics that relate to karyotypic instability. In the chronic phase, approximately 60% of CML patients show reduced telomere length without highly elevated telomerase activity or microsatellite alterations, indicating that telomere reduction may be linked to cell replication. Therefore, the Ph translocation might be a first event to immortalize cell proliferation. In the blast phase, 50% of CML patients have high levels of elevated telomerase activity and the same number of patients had microsatellite changes. Of note is that most patients with telomerase up regulation in the blast phase had additional cytogenetic changes and >60% of them showed microsatellite changes at least at one locus. In contrast, most patients without telomerase activity did not show microsatellite changes. These findings may indicate that telomerase up-regulation in the blast phase of CML patients is closely associated with microsatellite changes (representative of genome instability), while blast cells in the remaining patients (30%) maintain their proliferative capability without microsatellite changes and telomerase up regulation. This further suggests that there is also an unknown mechanism for genome stability without the process of telomerase up-regulation in some patients with CML in blast crisis. PMID- 11426629 TI - Epstein-Barr virus-infected natural killer cell leukemia. AB - Natural killer cell leukemia (NK leukemia) is an aggressive form of lymphoproliferative disease of granular lymphocytes, and frequently complicates fulminant hemophagocytic lymphohistiocytosis. NK leukemia cells usually possess a single episomal form of Epstein-Barr virus (EBV), and therefore originate from a single EBV-infected NK cell. The NK leukemia cells lack endogenous Bcl-2 expression and are sensitive to apoptotic cell death. However, they constitutively produce interferon-gamma and maintain their survival in an autocrine fashion. The interferon-gamma released from NK leukemia cells may trigger the occurrence of hemophagocytic lymphohistiocytosis through activating macrophages/histiocytes. In the primary infection of EBV, T cells infected with the episomal form of EBV sometimes produce a high amount of interferon-gamma that may lead to the occurrence of hemophagocytic lymphohistiocytosis. Thus, it is important to determine the role of EBV in the increased production of interferon gamma that occurs in EBV-infected T and NK cells to clarify the developmental mechanism of NK leukemia and its paraneoplastic hemophagocytic lymphohistiocytosis. PMID- 11426630 TI - Disease features in acute myeloid leukemia with t(8;21)(q22;q22). Influence of age, secondary karyotype abnormalities, CD19 status, and extramedullary leukemia on survival. AB - Over a period of 14 years, 50 patients (12 children and 38 adults) of whom 46 had acute myeloid leukemia (AML) and 4 had myelodysplastic syndrome characterized by the t(8;21)(q22;q22) translocation were referred to the Royal Marsden Hospital. The clinicopathological features of these cases were analyzed to determine the influence of age, secondary karyotype abnormalities, and expression of the lymphoid marker CD19 on event free survival, and presence of extramedullary leukemia on overall survival. They were treated with a variety of chemotherapy protocols and some had bone marrow transplantation. There appeared to be no difference in survival between children (age <17 years) and adults (age >16 years). Out of the 50 cases, 16 (32%) had the (8;21) translocation alone, 17 (34%) had additional loss of a sex chromosome and the remaining 17 (34%) had other karyotype abnormalities of which deletion or translocation of the long arms of a #9 was most common (observed in 8 of the 17 patients). The karyotype groups had a significant impact on survival, the group with loss of a sex chromosome having a poorer outcome and the group with abnormalities of chromosome 9 having a better outcome. CD19 positivity was seen in 21 of the 33 cases (63%) in whom it was measured compared to 11% observed in controls with AML without a t(8;21). CD19 status did not exert any influence on event free survival. Extramedullary leukemia (EML) occurred in 5 of the 50 cases (10%). In one patient it was observed at diagnosis but in the others it presented concurrent with bone marrow relapse. The overall survival of patients with EML was worse than that of the other patients but did not achieve statistical significance and was probably adversely affected by other factors. PMID- 11426631 TI - Experience with 9-cis retinoic acid in patients with relapsed and refractory non Hodgkin's lymphoma. AB - We conducted a phase II study to determine the efficacy and toxicity of 9-cis retinoic acid (9-cis RA), a pan-retinoid receptor agonist, in the treatment of patients with relapsed and refractory NHL. Patients were eligible if they had histologically documented relapsed or refractory T cell or indolent B cell NHL. The first three patients enrolled received 70 mg/m2 of 9-cis RA orally twice a day, but the remaining patients received a single oral daily dose of 100 mg/m2. After 6 weeks of therapy, tumor response was assessed objectively. Response rate and toxicity were determined in all 29 eligible patients based on an intent-to treat analysis. Four patients (14%) responded (3 PRs and 1 CR; 95% CI 4%-33%). One patient had a minor response, and eight had stable disease. Responses were observed in two (11%) of 19 patients with B-cell lymphoma and in two (20%) of 10 patients with T-cell lymphoma. The median time-to-treatment failure for the 29 eligible patients was 8 weeks. The most frequent toxic effects were dry skin, headache, hypertriglyceridemia, and hypercalcemia. Five patients discontinued therapy due to toxic side effects, but no toxic deaths occurred during the study. We conclude that 9-cis RA has a modest activity in relapsed and refractory NHL. In this study, responses were observed in patients with B-cell lymphomas and those with T-cell lymphomas. PMID- 11426632 TI - OPP-EBV-CAD regimen as salvage treatment in advanced refractory or resistant multiple myeloma. AB - With the aim of developing an effective therapy for heavily pretreated refractory MM outpatients, we evaluated the OPPEBVCAD regimen, a Hodgkin's disease-derived protocol that includes many drugs effective in MM administered in a sequential schedule. Twenty-two pts aged 42-72 years, with symptomatic highly-pretreated refractory (18 cases), or primary resistant MM (four cases. including two pts with plasma cell leukemia-PCL) received this therapy every 28 days (2-4 cycles, followed by a maintenance program). Therapeutic response (Chronic Leukemia Myeloma Task Force criteria) and performance status (PS) and pain (W.H.O.) were evaluated. All of the pts were evaluable for response. There were 9 (40%) objective responses (OR: stabilization of blood counts and bone lesions, serum calcium normalization, 50% or more reduction in the concentration of serum monoclonal component (MC), 90% reduction in Bence-Jones proteinuria), 8 (36%) partial responses (PR: 25-50% reduction in serum MC), 1 no response or stable disease (NR), and 4 (18%) cases of progressive disease (PD). OR plus PR were 77%. Of the 4 primary resistant tumors (2 PCL and 2 MM), 2 achieved PR, 1 OR (a PCL case) and 1 progressed. Median survival was 15 months for responding pts (OR plus PR) and 4.5 months for non-responders (NR plus PD). PS and pain improved in 15 pts and did not change in 9. The most frequent side effects were cytopenias, with one drug related infective death. The OPPEBVCAD regimen proved to be an effective therapy for refractory relapsing or primary resistant MM: in responders (two thirds of the pts), survival was prolonged by about 10 months. Its efficacy in anthracycline-treated pts, as well as the feasibility of using it on an outpatient basis without any continuous drug infusions, make this regimen a promising third line salvage therapy. PMID- 11426633 TI - A prospective randomized study of Chop versus Chop plus alpha-2B interferon in patients with intermediate and high grade non-Hodgkin's lymphoma: the International Oncology Study Group NHL1 Study . AB - The addition of a brief alpha interferon regimen to each CHOP induction cycle, plus one year of alpha interferon thrice weekly maintenance therapy, has no early effect on response rates or survival in patients with Intermediate or High grade cell NHL. BACKGROUND: The CHOP (Cyclophosphamide, Adriamycin. Vincristine, Prednisone) regimen is the most widely used first-line therapy for patients with Intermediate or High Grade (IG/HG) non-Hodgkin's lymphoma (NHL). Alpha 2b interferon (INF) enhances response rates and improves survival in low-grade NHL. The International Oncology Study Group (IOSG) conducted a prospective randomized study comparing CHOP alone or combined with INF in patients with IG/HG-NHL. The primary study aim was to compare the objective response rates in these patient cohorts. PATIENTS AND METHODS: Patients with a confirmed diagnosis of measurable NHL of International Working Formulation (IWF) groups D to H histology were randomized to receive CHOP alone or CHOP with 5Mu INF s.c. for 5 days on days 22 to 26 of each 28 day cycle with INF 5 million units (Mu) given three times per week subcutaneously for 52 weeks in those patients who responded to CHOP plus INF. RESULTS: The overall response rates were equivalent in both groups: CHOP alone (214 patients) 81% (complete 55%, partial 26%); CHOP plus INF (221 patients) 80% (complete 54%, partial 26%). At 36 months, the actuarial survival rate was equivalent in both groups. CONCLUSIONS: There is no apparent early advantage in terms of response or survival conferred by adding the study INF regimen to CHOP therapy for patients with IG/HG-NHL. PMID- 11426634 TI - Surgical technique for combined intestine-auxiliary liver transplantation in rats: development of a new microsurgical model. AB - Combined liver-intestine transplantation is an evolving procedure, and auxiliary liver transplantation has several advantages over standard orthotopic liver transplantation. We present a new model of combined intestine-auxiliary liver transplantation in rats. Total small bowel and 60% liver were harvested en bloc. An aortic segment that contained the celiac axis and superior mesenteric artery ensured blood supply to the graft. Venous drainage of the grafted intestine was achieved via the intact portal vein of the graft. The infrahepatic vena cava was cut at different levels during the modification period and at the oblique level of the left renal vein in consecutive series. Revascularization was accomplished by end-to-side anastomosis of the aorta and of the infrahepatic vena cava. The recipient small bowel was resected and the intestine continuity restored by anastomosis. Total operation time averaged 130 min. The overall survival rate of 3 months in the consecutive series was 80% (16/20). Exploratory laparatomy and histologic study in 3 rats on 90 days after transplantation revealed normal and viable grafts. Liver function was normal and both grafted liver and intestine showed normal histologic architectures in 5 rats observed for 12 months after transplantation. The present model is reproducible and allows preclinical research on several aspects of experimental combined intestine-auxiliary liver transplantation. PMID- 11426635 TI - Orthotopic intestinal transplantation using the cuff method in rats: a histopathological evaluation of the anastomosis. AB - Segmental small intestine transplantation (SIT) in rats, using a cuff technique, has achieved a high success rate. However, there have been few reports on the influence of the foreign body reaction to polyethylene cuff on vessel anastomoses and graft after SIT. This study involves the histopathological examination of the site of cuff anastomosis and grafts in the short- and long-term survival of segmental SIT. The data obtained from the suture anastomosis model also served as a control. One week after heterotopic segmental SIT using the cuff technique, orthotopic continuations were carried out in syngeneic combination. Twenty-five of 30 rats surviving >200 days (83.3%) were examined for vessel anastomosis. All arterial anastomoses were patent, but the portovenous anastomoses in 10 grafts (33%) were totally occluded and were associated with the formation of collateral vessels. Histopathological examination demonstrated good patency of the artery and vein anastomotic site in the short term, but granulation, fibrosis, and neovascularization at the anastomosis site surrounding the cuffs in the long surviving group. However, the grafts appeared to be intact, with normal features of the villi. On the contrary, the site of the sutured anastomosis in the long survival rats showed no inflammatory reaction. Although a polyethylene cuff caused foreign body reaction, the graft blood supplies were maintained by collateral vessels. Considering the low mortality and high success rate, polyethylene cuff is good for short-term study and an alternative method for long term SIT experiments. PMID- 11426636 TI - Simplified method of heterotopic rat heart transplantation using the cuff technique: application to sublethal dose protocol of methotrexate on allograft survival. AB - Rodent heterotopic heart transplantation (HHT) models have been developed for the study of transplantation immunology. Most of these transplantations are performed by hand-suture techniques, requiring several months of training. We describe a modified technique of rat HHT in the neck, using a cuff method that can be mastered by beginners within a few weeks. Our main modification of the rat HHT in the neck is that the right superior vena cava of the graft is chosen as an outflow duct, while the pulmonary artery has been taken as an effluent drainage in the ordinary HHT models. The aorta of the donor is anastomosed with the carotid artery of the recipient. Donation can be completed within 5 min and vascular connections in the recipient done within 3 min, resulting in a minimum of ischemia time. Using this minimum surgical intervention model, we tested the immunosuppressive effect of a sublethal dose of methotrexate (MTX), which has been widely used in cancer therapy. Our results showed that high doses of MTX severely suppressed the recipient bone marrow, but prolonged heart allografts for more than 365 days after HHT. In conclusion, the new model simplified the rat HHT procedure and made it possible for the beginner of rodent transplantation to master this skill within a few weeks. Using this minimized intervention technique, we found that the high doses of MTX can significantly prolong the survival of fully mismatched DA heart graft in PVG/c recipient. PMID- 11426637 TI - Computer-guided microsurgery: surgical evaluation of a telerobotic arm. AB - A new telerobotic arm was evaluated in different microsurgical procedures. The system consisted of a robotic arm bearing a microsurgical forceps controlled by a joystick-like master unit operated by the surgeon's hand. The robotic arm functioned as an operating as well as assisting instrument. Advantages of the system were its precise functioning, which was especially important when tissue or instruments had to be held for a longer period of time, as well as its ability to replace an assisting person to some extent. Deficiencies of the system were its 10-min startup, the spatial conflict with in the operative field, and the poor rotation of the robotic tip. In some procedures, the telerobotic arm could replace the standard microsurgical instrument guided by the surgeon's second hand; in other maneuvers it could function as the surgeon's third hand with precision and void of tremor. PMID- 11426638 TI - Cell proliferation and death of growth plate chondrocyte caused by ischemia and reperfusion. AB - The aim of this study was to assess the short-term response of cell kinetics of growth plate chondrocytes under conditions of warm ischemia and reperfusion. To understand the time-course changes that occur after reperfusion, 0 and 6 h of warm ischemia was produced in the right hindlimb of 35-day-old Wistar rats by isolating the vascular pedicle occlusion. The animals were killed at 12, 24, 48, or 96 h postoperatively after reperfusion, and proximal tibia growth plates were investigated. To investigate the effect of the ischemia period on the kinetics of growth plate chondrocytes, 0, 2, 4, 6, and 8 h of ischemia was induced, and the animals were killed for evaluation 24 h after reperfusion. For evaluation of cell kinetics, BrdU was used to observe the changes in cell proliferation of growth plate chondrocytes, and TUNEL was used to estimate the changes in rate of cell death. In the time-course study, both 0 and 6 h of ischemia increased cell proliferation at 12 and 24 h after reperfusion; however, at 48 and 96 h, the proliferation rate was not further increased. At 12 and 24 h postoperatively, 6 h of ischemia increased chondrocyte proliferation more than 0 h of ischemia with significant differences; 6 h of ischemia led to an increased cell death rate at 12, 24, and 48 h postoperatively, whereas 0 h of ischemia did not affect the cell death rate. In the ischemia time-dependent study, the cell proliferation rate induced by 4 h of ischemia was highest in all controlled periods of ischemia. Cell death rate increased gradually with increases in ischemia time 24 h after reperfusion. This experiment showed that ischemic damage causes short-term postoperative changes in the kinetics of growth plate chondrocytes. PMID- 11426639 TI - A Schwann cell-seeded intrinsic framework and its satisfactory biocompatibility for a bioartificial nerve graft. AB - To optimize the internal environment of a collagen nerve tube, we designed a Schwann cell-seeded intrinsic framework and its biocompatibility was investigated. We fixed 6-0 polyglactin woven filaments (Vicryl) or polydioxanone monofilaments (PDS) on a silicone ring in a net fashion. It was coated with matrigel and then incubated with cultured newborn or adult Schwann cells. Furthermore, we implanted 1.5-cm-long filament-filled collagen tubes in a rat model. Using a live/dead fluorescent assay and electron microscopy, we found that adherent Schwann cells onto filaments remained viable and oriented longitudinally along filaments. The preliminary in vivo study indicated that a mild inflammatory reaction was present around the tube wall. However, nerve regeneration occurred around and between filaments. We concluded that the arrangement of Schwann cell columns onto filaments was achieved, mimicking Bunger bands. It was shown that the biomaterials did not impede nerve regeneration. PMID- 11426640 TI - The EGF receptor family as targets for cancer therapy. AB - Human carcinomas frequently express high levels of receptors in the EGF receptor family, and overexpression of at least two of these receptors, the EGF receptor (EGFr) and closely related ErbB2, has been associated with a more aggressive clinical behavior. Further, transfection or activation of high levels of these two receptors in nonmalignant cell lines can lead to a transformed phenotype. For these reasons therapies directed at preventing the function of these receptors have the potential to be useful anti-cancer treatments. In the last two decades monoclonal antibodies (MAbs) which block activation of the EGFr and ErbB2 have been developed. These MAbs have shown promising preclinical activity and 'chimeric' and 'humanized' MAbs have been produced in order to obviate the problem of host immune reactions. Clinical activity with these antibodies has been documented: trastuzumab, a humanized anti-ErbB2 MAb, is active and was recently approved in combination with paclitaxel for the therapy of patients with metastatic ErbB2-overexpressing breast cancer; IMC-C225, a chimeric anti-EGFr MAb, has shown impressive activity when combined with radiation therapy and reverses resistance to chemotherapy. In addition to antibodies, compounds that directly inhibit receptor tyrosine kinases have shown preclinical activity and early clinical activity has been reported. A series of phase III studies with these antibodies and direct tyrosine kinase inhibitors are ongoing or planned, and will further address the role of these active anti-receptor agents in the treatment of patients with cancer. PMID- 11426641 TI - Design of growth factor antagonists with antiangiogenic and antitumor properties. AB - This review describes our recent efforts in the development of novel therapies for cancer. Our primary approach is to design synthetic agents that antagonize the function of growth factors that are critically involved in oncogenesis and angiogenesis. We achieve this by designing synthetic molecules that can recognize the exterior surface of the growth factor and so block the interaction with its receptor tyrosine kinase. A key step is the construction of synthetic agents that contain a large (> 400A2) and functionalized surface area to recognize a complementary surface on the target growth factor. In the course of this work we have discovered a molecule, GFB-111, that binds to PDGF, prevents it from binding to its receptor tyrosine kinase, blocks PDGF-induced receptor autophosphorylation, activation of Erk1 and Erk2 kinases and DNA synthesis. The binding affinity for PDGF is high (IC50=250 nM) and selective over EGF, IGF-1, aFGF, bFGF and HRGbeta. In nude mouse models GFB-111 also shows significant inhibition of tumor growth and angiogenesis. PMID- 11426642 TI - From oncogene to drug: development of small molecule tyrosine kinase inhibitors as anti-tumor and anti-angiogenic agents. AB - The confluence of two distinct but related activities in the past 10 years has dramatically accelerated efforts towards the discovery and development of novel drugs to treat cancer. The first is a rapidly emerging understanding that a number of distinct tyrosine kinases play roles in diverse but fundamentally important aspects of tumor progression (growth, survival, metastasis and angiogenesis). The second is the discovery that small molecule compounds have the capacity to potently and selectively inhibit the biochemical function of tyrosine kinases by competing for ATP binding at the enzyme catalytic site. These observations have been conjoined in major efforts to bring forward into clinical development novel cancer drugs with the potential to provide both clinical efficacy and improved tolerability. The focus of this review is on the development of small molecule tyrosine kinase inhibitors, and does not extend to other approaches that could be applied to disrupt the same pathways in clinical tumors (receptor and/or ligand-competitive antibodies, intrabodies, antisense ribonucleotides, ribozymes, phosphatase inhibitors or SH2/SH3-directed agents). Selected tyrosine kinase inhibitors, known or believed to be in development in cancer treatment trials, are summarized as are some of the key issues that must be addressed if these compounds are to be developed into clinically useful cancer chemotherapeutic agents. PMID- 11426643 TI - Farnesyltransferase and geranylgeranyltransferase I inhibitors and cancer therapy: lessons from mechanism and bench-to-bedside translational studies. AB - In 1990, more than 10 years after the discovery that the low molecular weight GTPase Ras is a major contributor to human cancer, farnesylation, a lipid posttranslational modification required for the cancer-causing activity of Ras, emerged as a major target for the development of novel anticancer agents. However, it took only 5 years from 1993, when the first farnesyltransferase inhibitors (FTIs) were reported, to 1998 when results from the first phase I clinical trials were described. This rapid progress was due to the demonstration of outstanding antitumor activity and lack of toxicity of FTIs in preclinical models. Although, many FTIs are currently in phase H and at least one is in phase III clinical trial, the mechanism of FTI antitumor activity is not known. In this review a brief summary of the development of FTIs as antitumor agents will be given. The focus of the review will be on important mechanistic and bench-to bedside translational issues. Among the issues that will be addressed are: evidence for and against inhibition of the prenylation of Ras and RhoB proteins in the mechanism of action of FTIs; implications of the alternative prenylation of K-Ras by geranylgeranyl-transferase I (when FTase is inhibited) in cancer therapy; GGTase I inhibitors (GGTIs) as antitumor agents; effects of FTIs and GGTIs on cell cycle machinery and progression and potential mechanisms by which FTIs and GGTIs induce apoptosis in human cancer cells. A thorough discussion about bench-to-bedside issues relating to hypothesis-driven clinical trials with proof-of-principle in man will also be included. This section will cover issues relating to whether the biochemical target (FTase) is inhibited and the level of inhibition of FTase required for clinical response; are signaling pathways such as H-Ras/PI3K/Akt and/or K-Ras/Raf/MEK/Erk relevant biological readouts?; is Ras (particularly N-Ras and H-Ras) mutation status a good predictor of clinical response?; in phase I trials should effective biological dose, not maximally tolerated dose, be used to determine phase II dose?; and finally, in phase II/III trials what are the most appropriate clinical end points for anti-signaling molecules such as FTIs? Parts of this topic have been recently reviewed (Sebti and Hamilton, 2000c). PMID- 11426644 TI - Development of anticancer drugs targeting the MAP kinase pathway. AB - Since the discovery of the role of ras oncogenes in tumorigenesis, we have witnessed an explosion of research in the signal transduction area. In the quest to understand how Ras transmits extracellular growth signals, the MAP kinase (MAPK) pathway has emerged as the crucial route between membrane-bound Ras and the nucleus. The MAPK pathway encompasses a cascade of phosphorylation events involving three key kinases, namely Raf, MEK (MAP kinase kinase) and ERK (MAP kinase). This kinase cascade presents novel opportunities for the development of new cancer therapies designed to be less toxic than conventional chemotherapeutic drugs. Furthermore, as a signal transduction-based approach to cancer treatment, inhibition of any one of these targets has the potential for translational pharmacodynamic evaluation of target suppression. The rationale for targeting the MAP kinase pathway will be reviewed here along with a discussion of various pharmacological approaches and the promise they hold for a new generation of anticancer drugs. PMID- 11426645 TI - Small molecule modulators of cyclin-dependent kinases for cancer therapy. AB - The majority of human malignancies have aberrancies in the Retinoblastoma (Rb) pathway. Loss in Rb function results from the phosphorylation and inactivation of Rb by the cyclin-dependent kinases (cdks), main regulators of cell cycle progression. Thus, modulators of cdks may have a role in the treatment of human malignancies. Flavopiridol, the first cdk modulator tested in clinical trials, demonstrates interesting preclinical features: cell cycle block, induction of apoptosis, promotion of differentiation, inhibition of angiogenic processes and modulation of transcriptional events. Initial clinical trials with infusional flavopiridol demonstrated activity in some patients with lymphomas and renal, colon gastric carcinomas. Main side effects were diarrhea and hypotension. Phase 2 trials with infusional flavopiridol, other schedules and combination with standard chemotherapies are ongoing. The second cdk modulator tested in clinical trials, UCN-01, is a PKC inhibitor that can also modulate cdk activity. Similar to flavopiridol, UCN-01 blocks cell cycle progression and promotes apoptosis. Moreover, UCN-01 may abrogate checkpoints induced by genotoxic stress due to inhibition of chk1 kinase. The first clinical trial of UCN-01 demonstrated very prolonged half-life (approximately 600 h), due to high binding affinity of UCN-01 to the human alpha-1-acid glycoprotein. Main side effects were headaches, vomiting, hypoxemia and hyperglycemia. Clinical activity was observed in some patients with melanoma and lymphoma. Trials of shorter infusions of UCN-01 or in combination with standard chemotherapeutic agents are ongoing. Although several important basic and clinical questions remain unanswered, development of cdk modulators is a reasonable strategy for cancer therapy. PMID- 11426646 TI - Small molecule inhibitors of dual specificity protein phosphatases. AB - One hallmark of neoplasia is the deregulation of cell cycle control mechanisms, which is secondary to altered protein phosphorylation. Dual specificity protein phosphatases uniquely dephosphorylate both phosphoserines/threonines and phosphotyrosines on the same protein substrate. As a class they regulate intracellular signaling through the mitogen activated and stress activated kinases and govern cellular movement through G1/S and G2/M cell cycle checkpoints by affecting the activity of cyclin-dependent kinases. In particular, the Cdc25 phosphatases, which dephosphorylate cyclin-dependent kinases, are overexpressed in many human tumors and this increased expression is associated with a poor prognosis. In addition to expression levels, the intracellular activity of Cdc25 phosphatases is determined by their subcellular distribution and physical proximity to substrates. Small molecules that either inhibit the catalytic activity or alter the subcellular distribution of these dual specificity protein phosphatases could provide effective tools to interrogate the role of phosphorylation pathways and may afford new approaches to the management of cancer. PMID- 11426647 TI - STAT proteins: novel molecular targets for cancer drug discovery. AB - Signal Transducers and Activators of Transcription (STATs) are a family of cytoplasmic proteins with roles as signal messengers and transcription factors that participate in normal cellular responses to cytokines and growth factors. Frequently, however, abnormal activity of certain STAT family members, particularly Stat3 and Stat5, is associated with a wide variety of human malignancies, including hematologic, breast, head and neck, and prostate cancers. Application of molecular biology and pharmacology tools in disease-relevant models has confirmed Stat3 as having a causal role in oncogenesis, and provided validation of Stat3 as a target for cancer drug discovery and therapeutic intervention. Furthermore, a constitutively-active mutant form of Stat3 is sufficient to induce oncogenic transformation of cells, which form tumors in vivo. Constitutive activation of Stat3 signaling is accompanied by upregulation of cyclin D1, c-Myc, and Bcl-x, changes consistent with subversion of normal cellular growth and survival control mechanisms. Block of constitutive Stat3 signaling results in growth inhibition and apoptosis of Stat3-positive tumor cells in vitro and in vivo. The observed dependence of certain tumors on constitutive Stat3 signaling for growth and survival has wide implications for cancer therapy, offering the potential for preferential tumor cell killing. This review evaluates constitutive Stat3 activation as a 'cancer-causing' factor, and proposes a number of molecular strategies for targeting Stat3 signaling for therapeutic intervention. PMID- 11426648 TI - Bcl-2 family proteins as targets for anticancer drug design. AB - Bcl-2 family proteins are key regulators of programmed cell death or apoptosis that is implicated in many human diseases, particularly cancer. In recent years, they have attracted intensive interest in both basic research to understand the fundamental principles of cell survival and cell death and drug discovery to develop a new class of anticancer agents. The Bcl-2 family includes both anti- and pro-apoptotic proteins with opposing biological functions in either inhibiting or promoting cell death. High expression of anti-apoptotic members such as Bcl-2 and Bcl-XL commonly found in human cancers contributes to neoplastic cell expansion and interferes with the therapeutic action of many chemotherapeutic drugs. The functional blockade of Bcl-2 or Bcl-XL could either restore the apoptotic process in tumor cells or sensitize these tumors for chemo- and radiotherapies. This article reviews the recent progress in the design and discovery of small molecules that block the anti-apoptotic function of Bcl-2 or Bcl-XL. These chemical inhibitors are effective modulators of apoptosis and promising leads for the further development of new anticancer agents. PMID- 11426649 TI - Telomere maintenance mechanisms as a target for drug development. AB - The shortening of the telomeric DNA sequences at the ends of chromosomes is thought to play a critical role in regulating the lifespan of human cells. Since all dividing cells are subject to the loss of telomeric sequences, cells with long proliferative lifespans need mechanisms to maintain telomere integrity. It appears that the activation of the enzyme telomerase is the major mechanism by which these cells maintain their telomeres. The proposal that a critical step in the process of the malignant transformation of cells is the upregulation of expression of telomerase has made this enzyme a potentially useful prognostic and diagnostic marker for cancer, as well as a new target for therapeutic intervention for the treatment of patients with cancer. It is now clear that simply inhibiting telomerase may not result in the anticancer effects that were originally hypothesized. While telomerase may not be the universal target for cancer therapy, we certainly believe that targeting the telomere maintenance mechanisms will be important in future research aimed toward a successful strategy for curing cancer. PMID- 11426650 TI - Critical appraisal of the use of matrix metalloproteinase inhibitors in cancer treatment. AB - Experimental studies performed prior to 1990 led to the widely held belief that matrix metalloproteinases (MMPs) produced by cancer cells are of critical importance in tumor invasion and metastasis. Based on this evidence, the pharmaceutical industry produced several well tolerated, orally active MMP inhibitors (MMPIs) which demonstrated efficacy in mouse cancer models. Phase III clinical trials initiated in 1997-98 using marimastat, prinomastat (AG3340), and BAY 12-9566 alone or in combination with standard chemotherapy in patients with advanced cancers (lung, prostate, pancreas, brain, GI tract) have recently been reported; no clinical efficacy was demonstrated. Bayer and Agouron have discontinued their ongoing Phase III drug trials of MMPIs in advanced cancer. In retrospect, the failure of MMPIs to alter disease progression in metastatic cancer might have been anticipated since MMPs appear to be important in early aspects of cancer progression (local invasion and micrometastasis) and may no longer be required once metastases have been established. Our understanding of MMP pathophysiology in cancer has expanded considerably in the past 10 years. Current views indicate that: (1) most MMPs in tumors are made by stromal cells, not carcinoma cells; (2) cancer cells induce stromal cells to synthesize MMPs using extracellular matrix metalloproteinase inducer (EMMPRIN) and cytokine stimulatory mechanisms; and (3) MMPs promote cell migration and the release of growth factors sequestered in the extracellular matrix. MMPs have a dual function in tumor angiogenesis: MMP-2 and MT1-MMP are required in breaking down basement membrane barriers in the early stage of angiogenesis, while other MMPs are involved in the generation of an angiogenic inhibitor, angiostatin. In spite of considerable recent progress in identifying multiple roles of MMPs in disease, our understanding of MMP function in cancer is far from complete (see Table 1). Based on accumulated data, it is recommended that future MMPI trials focus on: (1) patients with early stage cancer; (2) the use of MMPIs along with chemotherapy; (3) the measurement of MMPs in tumor tissue and blood as a means of identifying patients who are more likely to respond to MMPI therapy; and (4) identification of biomarkers that reflect activation or inhibition of MMPs in vivo. PMID- 11426651 TI - Potential roles of antisense technology in cancer chemotherapy. AB - Antisense technology may play a major role in cancer chemotherapy. It is clearly a tool of exceptional value in the functionalization of genes and their validation as potential targets for cancer chemotherapy. Additionally, there is now substantial evidence that antisense drugs are safe, and a growing body of data showing activity in animal models of human disease including cancer, and suggesting efficacy in patients with cancer. In this article, I review the progress in the technology, the anticancer antisense drugs in development and potential roles that antisense technology might play. PMID- 11426652 TI - Replication-selective oncolytic adenoviruses: virotherapy aimed at genetic targets in cancer. AB - Replication-selective oncolytic adenoviruses represent a novel cancer treatment platform. Clinical studies have demonstrated the safety and feasibility of the approach, including the delivery of adenovirus to tumors through the bloodstream (Heise et al., 1999b; Reid et al., 1999; Nemunaitis et al., 1999). The inherent ability of replication-competent adenoviruses to sensitize tumor cells to chemotherapy was a novel discovery that has led to chemosensitization strategies. These data will support the further development of adenoviral agents, including second-generation constructs containing exogenous therapeuitc genes to enhance both local and systemic antitumoral activity (Heise and Kirn, 2000; Hermiston, 2000; Agha-Mohammadi and Lotze, 2000). In addition to adenovirus, other viral species are being developed including herpesvirus, vaccinia, reovirus and measles virus (Kirn, 2000a; Martuza, 2000; Norman and Lee, 2000; Mastrangelo et al., 2000; Coffey et al., 1998; Martuza et al., 1991; Kirn, 2000b; Lattime et al., 1996). Since intratumoral spread also appears to be a substantial hurdle for viral agents, inherently motile agents such as bacteria may hold great promise for this field (Low et al., 1999; Sznol et al., 2000). Given the unknown predictive value of in vitro cell-based assays and murine tumor model systems for the efficacy and therapeutic index of replication-selective oncolytic adenoviruses in patients, we believe that encouraging adenoviral agents must be tested in well-designed clinical trials as soon as possible. Only then can the true therapeutic potential of these agents be realized. PMID- 11426653 TI - ONYX-015 selectivity and the p14ARF pathway. AB - ONYX-015, dl1520, is an adenovirus that lacks the E1B 55K gene and therefore lacks the capacity to neutralize p53 during infection. This virus induces high levels of p53 and fails to grow efficiently in primary epithelial cells. However, it does replicate in many tumor cells, including those expressing wild-type p53. In these cells, ONYX-015 fails to induce active p53. This is because the pathway from Ela to p53 is disrupted through loss of p14ARF. We propose that high levels of Mdm2 activity resulting from loss of p14ARF, and high levels of Mdm2 protein resulting from activated Ras prevent accumulation of functional p53 during infection of tumor cells that retain wild-type p53. PMID- 11426654 TI - Dendritic cell vaccination for cancer therapy. AB - A growing list of defined tumor-antigens opens the way to antigen specific immunotherapy of cancer. However current approaches are often limited in their potential to induce an effective anti-tumor response. Dendritic cells (DC) are natural adjuvants for the induction of antigen specific T cell response. They have been successfully used in clinical pilot trials to induce tumor specific immunity as well as clinical response in selected patients. Current research focuses on optimization of DC source, choice of antigen, antigen loading, mode of injection, as well as immuno-monitoring. Finally, a variety of immune escape mechanisms are operative at the tumor site and have to be overcome for successful vaccination. PMID- 11426655 TI - The rapamycin-sensitive signal transduction pathway as a target for cancer therapy. AB - The high frequency of mutations in cancer cells which result in altered cell cycle regulation and growth signal transduction, conferring a proliferative advantage, indicates that many of these aberrant mechanisms may be strategic targets for cancer therapy. The macrolide fungicide rapamycin, a natural product with potent antimicrobial, immunosuppressant, and anti-tumor properties, inhibits the translation of key mRNAs of proteins required for cell cycle progression from G1 to S phase. Rapamycin binds intracellularly to the immunophilin FK506 binding protein 12 (FKBP12), and the resultant complex inhibits the protein kinase activity of a protein kinase termed mammalian target of rapamycin (mTOR). The inhibition of mTOR, in turn, blocks signals to two separate downstream pathways which control the translation of specific mRNAs required for cell cycle traverse from G1 to S phase. Blocking mTOR affects the activity of the 40S ribosomal protein S6 kinase (p70s6k) and the function of the eukaryotic initiation factor 4E-binding protein-1 (4E-BP1), leading to growth arrest in the the G1 phase of the cell cycle. In addition to its actions on p70s6k and 4E-BP1, rapamycin prevents cyclin-dependent kinase activation, inhibits retinoblastoma protein (pRb) phosphorylation, and accelerates the turnover of cyclin D1 that leads to a deficiency of active cdk4/cyclin D1 complexes, all of which can inhibit cell cycle traverse at the G1/S phase transition. Both rapamycin and CCI-779, an ester analog of rapamycin with improved pharmaceutical properties and aqueous solubility, have demonstrated impressive activity against a broad range of human cancers growing in tissue culture and in human tumor xenograft models, which has supported the development of compounds targeting rapamycin-sensitive signal transduction pathways. CCI-779 has completed several phase I clinical evaluations and is currently undergoing broad disease-directed efficacy studies. The agent appears to be well tolerated at doses that have resulted in impressive anti-tumor activity in several types of refractory neoplasms. Important challenges during clinical development include the definition of a recommended dose range associated with optimal biological activity and maximal therapeutic indices, as well as the ability to predict which tumors will be sensitive or resistant to CCI 779. PMID- 11426656 TI - New agents in cancer clinical trials. PMID- 11426657 TI - Novel macrolactins as antibiotic lactones from a marine bacterium. AB - Seven new macrolactins (named G~M) and known macrolactins A and F were isolated from a culture broth of Bacillus sp. PP19-H3. The strain had been isolated from the macroalga, Schizymenia dubyi. Macrolactin A, which was 24-membered lactone, had previously been reported to show antibacterial, cytotoxic and antiviral activities. The new macrolactins include 22-membered ring or dicyclic lactone in addition to geometric isomers of known macrolactins A and F. The antibacterial activities of all the macrolactins examined in this study were relatively weak. PMID- 11426658 TI - New type II manumycins produced by Streptomyces nodosus ssp. asukaensis and their biosynthesis. AB - Five new type II manumycins, containing the hydroxyquinol mC7N unit, asukamycins A-II, B-II, C-II, D-II, E-II, were discovered in cultures of Streptomyces nodosus ssp. asukaensis. The biosynthetic origin of the type II manumycins from the type I compounds, containing an epoxyquinol mC7N unit, was deduced from the time course of production and proven by preparing [7'-13C]asukamycin A and demonstrating its incorporation into asukamycin A-II. PMID- 11426659 TI - Phagocytosis of alveolar macrophages after conagenin injection to rats. AB - Phagocytic functions of rat alveolar macrophages (AM) following intraperitoneal injection of conagenin (CNG) and of AM sub-populations fractionated by Percoll discontinuous gradient centrifugation were investigated. Phagocytosis of opsonized-sheep red blood cells (SRBC) following in vitro incubation with CNG showed a significant increase in a higher density of AM (fraction IV). In addition, phagocytosis was also increased in lower density ones (fractions I and II) by macrophage-activating factor (MAF) co-cultivation. CNG-injected rats for 5 consecutive days showed a dose-dependent increase in phagocytosis of AM compared to the control rats. Although the distribution of AM sub-population in rats injected CNG was not significantly different compared to the control rats, phagocytosis was significantly increased in AM of a lower density fraction (fraction II). These results suggest that CNG directly increases phagocytosis of AM in a higher density fraction, and indirectly enhances phagocytosis in AM of a lower density fraction via increasing MAF-like material production. PMID- 11426660 TI - The chemical structure of mumbaistatin, a novel glucose-6-phosphate translocase inhibitor produced by Streptomyces sp. DSM 11641. AB - The characterization of the structure of mumbaistatin (1), an effective inhibitor of the glucose-6-phosphatase system (EC 3.1.3.9), is reported. Isolation of mumbaistatin from cultures of Streptomyces sp. DSM 11641 was achieved by anion exchange and reversed-phase chromatography. The acid-labile inhibitor was methylated for the structure determination. Single-crystal X-ray structure analysis of a triply methylated dehydration product, C31H24O11, revealed the structure of an aromatic dispirodiketal (2), a compound containing a previously undescribed ring system. Extensive 2D-NMR experiments with mumbaistatin and with the methylation products showed that mumbaistatin itself possesses the hydroxydiketodicarboxylic acid structure 1, C28H20O12, which, in the presence of acid or upon activation through methyl ester formation, undergoes self condensation with loss of water to the dispirodiketal form (2). Mumbaistatin is an anthraquinone derivative, whose open-chain diketo form acts as a specific and powerful inhibitor of glucose-6-phosphate translocase: IC50=5 nM. The activity towards the same enzyme of the cyclized dispirodiketal derivatives is roughly one thousand times lower. PMID- 11426661 TI - Studies on anti-MRSA parenteral cephalosporins. IV. A novel water-soluble N phosphono type prodrug for parental administration. AB - A systematic approach for improving the water-solubility of anti-MRSA (methicillin-resistant Staphylococcus aureus) cephalosporin derivatives is described. We first tried to improve the water-solubility of 7beta-[2-(5-amino 1,2,4-thiadiazol-3-yl)-2(Z)-fluoromethoxyiminoacetamido]-3-[(E)-2-(1 methylimidazo[1,2-b]pyridazinium-6-yl)thiovinyl]-3-cephem-4-carboxylate (1a) by substitution of the C-3' pharmacophore. Replacement of the C-3' pharmacophore with a 1-methyl-4-pyridinio group improved the water-solubility without decreasing the anti-MRSA activity. Furthermore, we applied the N-modified prodrug strategy to the C-7 acyl group in order to enhance the water-solubility drastically. Among the compounds prepared, the N-phosphono type prodrugs 2a(1 methylimidazo[1,2-b]pyridazinium derivative) and 2b (1-methyl-4-pyridinio derivative) showed water-solubility appropriate for a product intended for intravenous injection and in vivo anti-MRSA activity comparable to that of vancomycin. PMID- 11426662 TI - Thiopeptide non-producing Streptomyces species carry the tipA gene: a clue to its function. PMID- 11426663 TI - Selective cytotoxicity and stereochemistry of aspochalasin D. PMID- 11426664 TI - Convergent synthesis of arisugacin skeletons and their acetylcholinesterase inhibitory activity. PMID- 11426665 TI - Determination of absolute stereochemistries of arisugacin F and Territrem B, novel acetylcholinesterase inhibitors. PMID- 11426666 TI - A rat model of liver cirrhosis and esophageal varices. AB - In an effort to develop a reproducible model of liver cirrhosis and esophageal varices, we administered phenobarbital (PhB) and carbon tetrachloride (CCl4) in 32 rats that had previously undergone complete devascularization of the left renal vein (DLRV). The operation was conducted to enhance the development of cephalad collaterals mainly responsible for the induction of esophageal varices. Thirty-two rats underwent sham operation and PhB and CCl4 administration, and 24 only sham-operated rats comprised the control group. After the induction of liver cirrhosis, histopathologic examination and morphometric analysis of the lower esophagus were performed to study the submucosal veins. We separately studied number of vessels in the submucosa, mean vessel area, percentage of submucosa occupied by vessels, and area of the single most dilated vein. All variables except the number of vessels were significantly higher in cirrhotic rats that had undergone DLRV (P<0.001). We conclude that induction of liver cirrhosis in rats that previously undergo complete DLRV could present a reproducible and reliable model for the induction of liver cirrhosis and esophageal varices. PMID- 11426667 TI - Homocyst(e)ine metabolism in hemodialysis patients treated with vitamins B6, B12 and folate. AB - Hyperhomocyst(e)inemia is commonly accepted as an independent atherosclerotic risk factor. In most hemodialysis patients, serum homocyst(e)ine is markedly elevated and may contribute to premature atherosclerosis in these patients. Whereas the beneficial effect of folate supplementation on serum homocyst(e)ine has been extensively studied, there are less detailed studies on the effects of cobalamin and pyridoxal phosphate alone, or in combination with folate. We examined the effect of a four-week course of intravenous treatment with folate (1.1 mg), cobalamin (1.0 mg), and pyridoxal phosphate (5.0 mg), administered once (group 1), twice (group 2) or thrice (group 3) weekly in 33 hemodialysis patients divided in three groups of 11 patients. All patients were followed for a further four weeks after treatment was stopped. Serum homocyst(e)ine, cobalamin, folate and pyridoxal phosphate, as well as the metabolites of homocyst(e)ine, methylmalonate, 2-methylcitrate and cystathionine, were determined before, during and after treatment. Baseline serum homocyst(e)ine correlated significantly with serum folate (P=0.0149), cobalamin (P=0.0047) and pyridoxal phosphate (P=0.0408). Correlations independent from the other metabolites or vitamins were found for methylmalonate (P=0.003) and folate (P=0.029). All regimens increased serum cobalamin significantly (in group 1 from 444 +/- 215 to 17,303 +/- 11,989 pg/ml, P<0.01; in group 2 from 542 +/- 633 to 44,896 +/- 15,772 pg/ml, P<0.001; in group 3 from 548 +/- 394 to 77,961 +/- 31,546 pg/ml, P<0.001), but did not increase any of the other vitamin levels. Serum homocyst(e)ine was lowered significantly by 39.8% +/- 31.9% (P<0.05) with two and by 30.1% +/- 26.9% (P<0.05) with three vitamin dosages weekly, but not with one dosage weekly. Since methylmalonate is known to be a sensitive marker of cobalamin deficiency, the data support an important influence of cobalamin levels on baseline homocyst(e)ine levels. Increasing cobalamin levels and additional treatment with folate and pyridoxal phosphate 156 may decrease serum homocyst(e)ine in the same way as high doses of folate alone. PMID- 11426668 TI - Risk of endotoxemia during the initial phase of gut decontamination with antimicrobial agents. AB - Decontamination of the digestive tract with antimicrobial agents has been used for prevention and therapy of bacterial translocation. With regard to the well described endotoxin-releasing properties of these agents, the question arises as to whether their enteral administration might result in an increased amount of intestinal endotoxins entering the circulation. Immunocompromised Wistar rats were intraduodenally challenged with live E. coli. Control animals received saline solution, decontaminated rats were treated with either tobramycin plus polymyxin B or ciprofloxacin alone through the duodenal tube. Plasma endotoxin activity and blood bacteria count were measured hourly over an observation period of 5 h. The intestinal bacterial count was determined at the end of the experiment. Gut decontamination in both groups receiving antimicrobial agents resulted in elevated plasma endotoxin levels compared with nondecontaminated controls. Maximum endotoxin levels were found to be 5-6 times higher in the ciprofloxacin group than in the control group and 2 times higher than in the tobramycin/polymyxin group. No positive blood cultures were detected. Intestinal bacterial count was similar in both treatment groups. Enterally applied antimicrobial agents bear an elevated risk of endotoxemia during the initial phase of gut decontamination. The amount of endotoxin translocating from the digestive tract to the circulation varies with the agents used. Polymyxin only partially reduced the observed endotoxin leakage from the gut. PMID- 11426669 TI - Relaxation induced by serotonin and sumatriptan in isolated guinea pig gallbladder strips. AB - The effects of 5-hydroxytryptamine (5-HT) and sumatriptan were investigated on isolated guinea pig gallbladder strips. While 0.1 microM-50 mM of 5-HT exhibited contractile and/or relaxant effects in quiescent preparations, the same concentrations of sumatriptan did not. On the other hand, carbachol-precontracted tissues were relaxed by the same amounts of 5-HT and sumatriptan in a concentration-dependent manner. The relaxant responses to 5-HT were not antagonized or changed by tetrodotoxin, indomethacin, capsaicin, NG-nitro-L arginine (L-NOARG), GR55562 [(+/-)-propranolol, 3-[3-(N,N-dimethylamino)propyl 1 4-hydroxy-N-[4-(pyridin-4-yl)phenyl]benzamide , S(-)-propranolol, methysergide, ketanserin. tropisetron, GR 113808 ([1-[2-(methylsulphonylamino)ethyl -4 piperidinyl]methyl-1-methyl-1H-indole-3-carboxylate maleate salt). pargyline, and fluvoxamine. The relaxant responses to sumatriptan were antagonized by GR55562 but not by S(-)-propranolol. These results suggest that 5-HT and sumatriptan cause relaxation in carbachol-precontracted isolated guinea pig gallbladder strips by yet undefined mechanisms. PMID- 11426670 TI - Biosynthetic response of cultured articular chondrocytes to mechanical vibration. AB - The objective of this study was to determine the effects of mechanical vibration loading on DNA and proteoglycan syntheses in cultured rabbit articular chondrocytes. Chondrocyte culture plates were placed in a vibratory apparatus and subjected to a mechanical vibratory load at various frequencies and periods in culture. Mechanical vibration was applied at a sinusoidal waveform of 1.4 g acceleration with frequencies of 200, 300, 400, 800, and 1600 Hz. 3H-Thymidine and H2(35)SO4 incorporation were used to detect radiolabeled DNA and proteoglycan syntheses, respectively. A frequency of 300 Hz showed a time-dependent augmentation of DNA synthesis and gave a maximal increase at day 3 with periodic vibration (8 h per day) and at 72 h or longer with continuous vibration. It also promoted proteoglycan synthesis in long-term culture (from 3 to 15 days) by periodic vibration. However. frequencies above 400 Hz suppressed biosynthesis. These results suggest that mechanical vibration at certain frequencies may modulate biosynthetic response of articular chondrocytes. PMID- 11426671 TI - Antithrombin III reduces renal ischemia-reperfusion injury in rats. AB - Few studies have pointed out the relationship between ischemia-reperfusion (IR) injury and the coagulation system. Antithrombin III (AT) has anti-inflammatory effects in IR injury. We investigated the effect of AT supplementation on renal IR injury in rats achieved by clamping of the left renal pedicle for 60 min and subsequent 24-h reperfusion after right nephrectomy. Sprague-Dawley rats were divided into three groups: sham-operated (no IR injury), ischemic controls, and an AT-treated group (250 U/kg before reperfusion). Creatinine values, tissue malondialdehyde (MDA) levels, myeloperoxidase (MPO) activity, and histopathological damage were investigated after 24 h of reperfusion. In addition, the 7-day survival rates in each group were evaluated. Creatinine and MDA levels and MPO activity were significantly elevated and histopathological damage was more severe in controls than in the sham group (P<0.05). Creatinine and MDA levels and MPO activity were significantly lower and there was less histopathological damage in the AT group than in controls. Accumulation of lipid peroxidation products and neutrophils were significantly inhibited by AT treatment. We conclude that AT may attenuate renal IR injury in rats. PMID- 11426672 TI - Effects of intraperitoneally administered vitamin C on antioxidative defense mechanism in rats with diabetes induced by streptozotocin. AB - We determined the effects of intraperitoneally administered vitamin C on the lipid peroxidation (as thiobarbituric acid-reactive substances, TBARS) and vitamin C and E levels and reduced glutathione (GSH) and glutathione peroxidase (GSH-Px) activity in the plasma, red blood cells (RBC), liver, and muscle of rats in relation to oxidative damage associated with diabetes induced by streptozotocin (STZ). One group was used as control and a second as diabetic. A third group received 30 mg vitamin C i.p. every other day. On day 4 after the injection of vitamin C, animals in the second and third groups were made diabetic by i.p. injection of STZ and administered vitamin C for 21 consecutive days, and we determined TBARS, vitamin E, and GSH levels and GSH-Px activities in plasma, RBC, liver, and muscle samples. Vitamin E levels in the plasma and liver were significantly higher (P<0.05) in the control group than in the diabetic group. Also, TBARS levels in the plasma, RBC, liver, and muscle samples were significantly lower (P<0.05) in controls than in the diabetic group. The TBARS levels in the RBC, liver, and muscle samples of the vitamin C group were significantly lower (P<0.05, P<0.01, and P<0.001, respectively). However, GSH-Px and GSH activities in RBC, liver, and muscle and vitamin C levels in liver were not significantly different between control and diabetic groups. Vitamin E levels in plasma (P<0.05, P<0.01) and liver (P<0.001), vitamin C levels in liver (P<0.001), and GSH (P<0.01) and GSH-Px activities in RBC (P<0.05, P<0.01) were significantly higher in the vitamin C group than both the control and diabetic groups. These results indicate that vitamin C has significant protective effects on the blood, liver, and muscle of rats against oxidative damage in diabetes. PMID- 11426673 TI - Effects of some nonsteroidal anti-inflammatory drugs on articular cartilage of rats in an experimental model of osteoarthritis. AB - We evaluate the chondrotoxic effects of some nonsteroidal anti-inflammatory drugs (NSAIDs) on articular cartilage in an experimental model of osteoarthritis (OA). Each of 40 Sprague-Dawley rats weighing 0.250 kg and 12 weeks old received weekly injections of sodium iodoacetate (1 mg/0.1 ml NaCl) in the right knees for 8 weeks to induce an experimental model of OA. The left knees served as controls. Four groups were formed of ten rats each. The control group received no medicine. In the other three groups, tiaprofenic acid, diclofenac, and indomethacin were given orally for 12 weeks. At the end of the 20th week, the complete groups were sacrificed and histologic evaluation performed. In the right knees of the control group, specific morphologic changes for OA were observed. Of the three NSAIDs tested, indomethacin was found to have deleterious effects on articular cartilage of both left and right knees. Diclofenac caused a statistically significant increase in the severity of most of the osteoarthritic parameters that were examined in the right knees, whereas tiaprofenic acid was observed to have some beneficial influences on the articular cartilage of right knees. Both diclofenac and tiaprofenic acid displayed the same deleterious effects on articular cartilage of left knees. We conclude that, in the prescription of NSAIDs for OA, it would be appropriate to choose a drug with a protective effect on chondrocytes and articular cartilage. PMID- 11426674 TI - Freeway management systems and motor vehicle crashes: a case study of Phoenix, Arizona. AB - This paper measures the impact of a freeway management system (FMS) on the incidence of reported motor vehicle crashes in Phoenix, AZ. Using a fixed effects negative binomial regression model, I find that the FMS reduces the frequency of crashes involving property damage only, possible injury, and minor injury by 25, 30, and 21%, respectively. I find no evidence for an effect on the frequency of major injury crashes or fatal crashes, although such accidents account for less than 5% of the total crashes in Phoenix. Classifying the data by the type of crash rather than by severity, I find that the frequencies of rearend crashes and sideswipe crashes are reduced by 25 and 37%, respectively, and I find no evidence of an effect on single vehicle crashes. The results are robust to many different model specifications, including a variety of functional forms, covariates, and data. A conservative estimate of the annual crash benefits of the FMS in Phoenix ranges from $4.8-13.2 million, depending on various assumptions about the value of pain and suffering and about the extent of crash underreporting. These annual crash benefits far outweigh the $1.6 million in annual operating and maintenance costs of the Phoenix FMS, and they offset considerably the approximately $47 million invested to date in the design and construction of the system. PMID- 11426675 TI - The epidemiology of traumatic injury-related fetal mortality in Pennsylvania, 1995-1997: the role of motor vehicle crashes. AB - METHODS: Rates and causes of traumatic injury-related fetal deaths in Pennsylvania were determined from a manual review of all fetal death certificates filed from 1995 to 1997 (7,131 cases). RESULTS: Thirty one traumatic injury cases were identified (6.5/100,000 live births). Most cases (94%) could be identified from the diagnosis code of 760.5 (maternal injury) and 87% contained narratives indicating specific injury mechanisms. Motor vehicles were the leading cause of injury (81%). Placental separation was the leading diagnosis (42%). CONCLUSIONS: The ICD-9-CM code 760.5 appears to be a specific indicator of traumatic fetal death, though it is not known how sensitive an indicator it is. Though not usually E-coded, the death certificates contained enough information to allow ascertainment of injury mechanism. These are very conservative estimates of the burden of the problem. The major role that motor vehicle injuries have on reported traumatic fetal injury deaths was shown and a significant new challenge for child passenger safety advocates is indicated. PMID- 11426676 TI - Public beliefs about the preventability of unintentional injury deaths. AB - This report is based upon the results of a national random digit dialed telephone survey in which 943 adults were queried. Subjects reported the proportion of deaths due to motor vehicle crashes, falls, fires/burns, drowning and poisoning that they felt were preventable. On average, respondents believed that 56% of 'fatal accidents' were preventable; as were 62% of motor vehicle crash deaths, 53% of fall deaths, 67% of drownings, 62% of fire/burn fatalities and 70% of accidental poisonings. Logistic regression models predicting preventability beliefs differed according to the type of injury event in question, but socio economic status and perceived alcohol involvement were positive predictors of beliefs related to all of the injuries under study. The ramifications of these findings and future research directions are discussed. PMID- 11426677 TI - Preventability of vehicle-related fatalities. AB - Several different methods have been used to evaluate the survivability of traffic injuries. Previously published methods were reviewed to develop a specific method for vehicle-related fatalities. All fatally injured victims (n = 474) of vehicle related crashes in the four northern-most counties of Sweden during a 5-year period were studied. Almost half (48%) of the victims had non-survivable injuries. The remaining cases were classified into different injury groups according to the Injury Severity Score (ISS) including 56 (12%) with an ISS < 25, 150 (32%) with ISS 25-49 and 42 (9%) with ISS 50-74. The median distance from the scene to the nearest hospital was 49 km. In the cases with ISS < 50 the medical care was further analyzed. In close to half of these cases, the victim did not receive optimal care in time with transportation time being of major importance. The absence of first aid was judged to have contributed to the death in 4% of the cases. PMID- 11426678 TI - Firearm availability and unintentional firearm deaths. AB - BACKGROUND: Between 1979 and 1997, almost 30,000 Americans died from unintentional firearm injuries, half of whom were under 25 years of age and 4,600 of whom were less than 15 years old. PURPOSE: To explore the association between state firearm levels and rates of unintentional firearm deaths by age group, accounting for several potential confounders. METHODS: The study used a proxy for firearm availability and pooled cross-sectional time-series data on unintentional firearm deaths for the 50 United States from 1979 to 1997. Negative binomial models were used to estimate the association between firearm availability and unintentional firearm deaths. RESULTS: A statistically significant and robust association exists between gun availability and unintentional firearm deaths for the US as a whole and within each age group. Multivariate analysis found that, compared to states with the lowest gun levels, states with the highest gun levels had, on average, 9 times the rate of unintentional firearm deaths. These results hold among men and women, for Whites and African Americans. CONCLUSION: Of the almost 30,000 people who died in unintentional firearm deaths over the 19-year study period, a disproportionately high number died in states where guns are more prevalent. The results suggest that the increased risk of unintentional violent death among all age groups is not entirely explained by a state's level of poverty, urbanization, or regional location. PMID- 11426679 TI - Roadway safety in rural and small urbanized areas. AB - Police Accident Reports (PAR) reveal that in a 5-year period between 1993 and 1997, there were 892 crashes at 87 two lane, undivided roadway sites in Strafford County, NH, a county consisting of suburban and rural communities. The purpose of this paper is to describe: (1) logistic regression model building efforts to identify statistically significant factors that predict the probabilities of crashes and injury crashes; and (2) to use these models to perform a risk assessment of the study region. The models are functions of factors that describe a site by its land use activity, roadside design, use of traffic control devices and traffic exposure. Comparative risk assessment results show village sites to be less hazardous than residential and shopping sites. Residential and shopping sites, which are distinctly different from village sites, reside in single purpose, land-use zones consisting mostly of single-family dwelling units and roadside shopping units with ample off-street parking. Village sites reside in multi-purpose, land-use zones permitting a combination of activities found in residential, shopping and commercial areas. They are pedestrian friendly, that is, have sidewalks and crosswalks, permit onstreet parking, have speed limits and other amenities that promote walking. Adjusted odds ratios and other comparative risk measures are used to explain why one site is more hazardous than another one. For example, the probability of a crash is two times more likely at a site without a sidewalk than at a site with one. The implications on roadway design to improve safety are discussed. PMID- 11426680 TI - Pickup truck use in the National Personal Transportation Survey. AB - To guide interventions to prevent injuries to pickup-truck occupants, we characterized pickup truck ownership, drivers and use in the 1995 National Personal Transportation Survey, which collects travel data from the civilian noninstitutionalized population of the US. SUDAAN software was used to account for the complex nature of the sample. Pickup truck ownership was more common in households with more vehicles, in rural households, in households living in single family homes and mobile homes, and in middle-income households. Among US regions, pickup truck ownership was highest among households in the mountain west. Pickup truck ownership was greater in households with two adults, whether or not children or youths were present, but this was largely due to the number of vehicles in these households. Driving a pickup on the sample day was more frequent among men, among drivers with less education and among full-time workers. A higher proportion of trips to work, work-related trips, longer trips and trips with fewer people were by pickup truck. Seat belt use was lower among pickup truck drivers than drivers of other vehicles. For only 0.5% of households (those with three or more members and a pickup truck as their only vehicle), restrictions on travel in cargo areas might be burdensome. Restrictions on cargo area travel, strengthening existing seat belt laws and social marketing strategies might increase the safety of pickup truck occupants. PMID- 11426681 TI - Evaluation of the effects of North Carolina's 0.08% BAC law. AB - This study was conducted to determine whether the lowered BAC limit for drivers in North Carolina resulted in fewer alcohol-related motor vehicle crashes. We used time-series analysis to examine several indicators of alcohol involvement in both injury and fatal crashes between 1991 and 1996. Data from NC crash files as well as the Fatality Analysis Reporting System (FARS) are used. We also examined several indicators used in previous research on lower BAC limits using a before after design to compare North Carolina with 37 states that did not change their BAC limit for 24 months before and after the BAC limit was lowered. When controlling for the pre-existing downward trend in drinking driver crashes, along with other pertinent factors such as amount of travel and number of weekends per month, there was no evidence of either a significant shift or a change in the downward trend of alcohol-related crashes associated with the lowered BAC limit. In conclusion, although the lower BAC limit was actively enforced and a substantial proportion of drinkers were aware of the new BAC limit, the drinking driving population in North Carolina, at the time the lower limit took effect in October 1993, was simply unresponsive to this change. PMID- 11426682 TI - The effects of driving cessation on the elderly with dementia and their caregivers. AB - This research explores how the loss of driving privileges by impaired drivers affects households. The particular focus is on the travel behavior and preceptions of people living in households where an elderly driver has had his or her license revoked due to Alzheimer's disease or a related dementia. The data for this analysis were drawn from a 1996 survey of households in California which queried the caregivers of people with dementia on how the former drivers access necessary destinations once they can no longer drive, and on the difficulties faced by other household members in seeking alternative means of transportation. After losing their license, the vast majority of people surveyed depended on informal support systems for transportation, such as rides from family and friends. Although such arrangements were not reported to be a problem for the majority of households, certain groups of non-drivers reported difficulty accessing services, particularly social and recreational destinations. The most commonly reported problem was a lack of available licensed drivers to chauffeur non-drivers. Importantly, no increase was observed in the number of people walking, using public transit, taxis, or van services following license revocation. People who did not live with at least one licensed driver and those who were younger and healthier reported the greatest mismatch between their need and desire to travel and the availability of transportation. In addition, some caregivers reported that they frequently missed work or stopped working entirely in order to care for and chauffeur people in the former drivers' household. Overall, these findings reinforce the importance of both developing transportation policies to support the functioning of informal transportation structures and in improving the range of alternative transportation options for those individuals with particular disabilities--like dementia--who are not well served by either informal arrangements or by formal transportation services for the disabled. PMID- 11426683 TI - Assessment of hospital and police ascertainment of automobile versus childhood pedestrian and bicyclist collisions. AB - Information about automobile versus pedestrian or bicyclist collisions differ according to the source of collision ascertainment. Hospital records and police reports focus on different characteristics of a collision, which reflects differences in case finding as well as information regarding risk factors. Pedestrians and bicyclists 0-14 years of age who were involved in a motor vehicle collision in the city of Long Beach, CA, between January 1, 1992 and June 30, 1995, were included in the study. Police reports and medical records were used to identify cases. Police reports were found for 1,015 cases, and medical records identified 474 cases; 379 cases were common to both sources. A capture-recapture model was used to evaluate the degree of overlap between the two sources and to derive "ascertainment corrected" injury rates. The injury rate from combined sources was 333.5/100,000 children per year, the pedestrian injury rate was 210.6/100,000 children per year and the bicyclist injury rate was 122.9/100,000 children per year. The "ascertainment corrected" injury rate overall was 381.3/100,000 children per year, 233.0/100,000 for pedestrians and 153.9/100,000 children per year for bicyclists. Eighty percent of hospital-reported cases were captured in the police database, whereas only 37% of police-reported cases were captured by the hospital database. We found that hospital sources identified younger children, fewer bicyclists, more Asian and Hispanic children, and fewer African-American children when compared to police sources. For more comprehensive surveillance resulting in more accurate incidence rates and more complete information, it is better to use multiple sources of data. PMID- 11426684 TI - In-vehicle distraction and fatal accidents in England and Wales. AB - The paper describes the coding and analysis of a database of police fatal accident reports to investigate the extent to which in-vehicle distraction is a contributory factor in vehicle crashes. A particular focus has been the involvement of mobile telephones and entertainment systems. Analysis of accidents occurring over the period 1985-1995 shows that in-vehicle distraction is reported as a contributory factor in about 2% of fatal accidents (although this figure may be a conservative estimate). Specific examples of distraction attributed to entertainment systems and telephones have been identified. Electronic driver information systems are also of particular interest, but have not featured in the available data. Work is progressing, on an annual cycle, to obtain, code and analyse further data and this is expected to provide an invaluable source of information for accident researchers. PMID- 11426685 TI - Causal models for road accident fatalities in Yemen. AB - An identification of the causes of road accident fatalities is becoming more important with the growth of technology, population, number of vehicles and the need for their use. Many authors have addressed the problem in the past but no universal findings have been obtained. The problem tends to be different under different environments and for different geographical regions. The aim of this paper is to develop a model for the analysis and forecasting of road accident fatalities in Yemen considering data restrictions. The proposed data has a particular structure of accident occurrence that has not been reported in any existing research using data in other countries. The available data for the period 1978-1995 is used to build models to understand the nature and extent of the causes of fatalities. Part of the data is used for model building and part of it for test purposes. The issues of correlation and causality have been addressed and multiple collinearity is investigated and dealt with. Two alternative models are proposed based on both statistical grounds and that of practicality in viable decision making. The influence of consuming a locally grown stimulant called Qat on road users has been addressed and it is found that it increases the risk of accidents. This is not the common understanding within the authorities in Yemen as growing and consuming Qat is unregulated. PMID- 11426686 TI - Safety and ergonomics evaluation of hybrid systems in Hong Kong. AB - This study is aimed at understanding common safety and ergonomics problems with robot installation in Hong Kong. Information was collected through direct observations and personal interviews with robot users from advanced hybrid manufacturing systems. The application areas of robots and types of safety devices employed were investigated. The ages, types of jobs, daily working hours and working conditions of personnel were examined. Fire precaution measures, types and varieties of training programs for users were analyzed. The results showed that although investments were made for installation of robots for improving production activity, limited attention was paid to the health, safety, and ergonomics aspects in the hybrid systems. There were also cases reported where no formal guidelines for robot manipulation and operation were provided. It seems that management and users paid attention to robot function and capability more than robot safety during robotization process. Conclusions were made by the authors that efforts should immediately be made to adopt recommendations on the safety devices and facilities and training requirements for improving users' health and safety before fatal accidents occur. PMID- 11426687 TI - Membrane fusion. AB - The fusion of biological membranes results in two bilayer-based membranes merging into a single membrane. In this process the lipids have to undergo considerable rearrangement. The nature of the intermediates that are formed during this rearrangement has been investigated. Certain fusion proteins facilitate this process. In many cases short segments of these fusion proteins have a particularly important role in accelerating the fusion process. Studies of the interaction of model peptides with membranes have allowed for increased understanding at the molecular level of the mechanism of the promotion of membrane fusion by fusion proteins. There is an increased appreciation of the roles of several independent segments of fusion proteins in promoting the fusion process. Many of the studies of the fusion of biological membranes have been done with the fusion of enveloped viruses with other membranes. One reason for this is that the number of proteins involved in viral fusion is relatively simple, often requiring only a single protein. For many enveloped viruses, the structure of their fusion proteins has certain common elements, suggesting that they all promote fusion by an analogous mechanism. Some aspects of this mechanism also appears to be common to intracellular fusion, although several proteins are involved in that process which is more complex and regulated than is viral fusion. PMID- 11426688 TI - Membrane fusion induced by phospholipase C and sphingomyelinases. AB - In the past decade lipid vesicle fusion induced by either bacterial PC-preferring phospholipase C, phosphatidylinositol-specific phospholipase C, sphingomyelinase, or a combination of phospholipase C and sphingomyelinase has been demonstrated. In the present paper, the experimental evidence is reviewed, and discussed in terms of the underlying molecular mechanisms of fusion, and of the possible physiological relevance of these findings. PMID- 11426689 TI - Mechanisms of initiation of membrane fusion: role of lipids. AB - Main emphasis in studies on the mechanisms of fusion of cellular membranes has been in the roles of various proteins, with far less interest in the properties of lipids. Yet, on a molecular level fusion involves the merging of lipid bilayers. Studies so far have revealed lipids forming inverted non-lamellar phases to be important in controlling membrane fusion. However, the underlying molecular level mechanisms have remained controversial. While this review is focused on presenting one possible mechanism, involving so-called extended lipid conformation, we are also advocating the view, that in order to obtain a more complete understanding of this process it is necessary to merge the relevant physicochemical properties of lipids with the models describing the specific functions of proteins. To this end, taking into account the central importance of fusion in a wide range of cellular processes, we may anticipate its control to open novel possibilities also for therapeutic intervention. PMID- 11426690 TI - Common properties of fusion peptides from diverse systems. AB - Although membrane fusion occurs ubiquitously and continuously in all eukaroytic cells, little is known about the mechanism that governs lipid bilayer fusion associated with any intracellular fusion reactions. Recent studies of the fusion of enveloped viruses with host cell membranes have helped to define the fusion process. The identification and characterization of key proteins involved in fusion reactions have mainly driven recent advances in our understanding of membrane fusion. The most important denominator among the fusion proteins is the fusion peptide. In this review, work done in the last few years on the molecular mechanism of viral membrane fusion will be highlighted, focusing in particular on the role of the fusion peptide and the modification of the lipid bilayer structure. Much of what is known regarding the molecular mechanism of viral membrane fusion has been gained using liposomes as model systems in which the molecular components of the membrane and the environment are strictly controlled. Many amphilphilic peptides have a high affinity for lipid bilayers, but only a few sequences are able to induce membrane fusion. The presence of alpha-helical structure in at least part of the fusion peptide is strongly correlated with activity whereas, beta-structure tends to be less prevalent, associated with non native experimental conditions, and more related to vesicle aggregation than fusion. The specific angle of insertion of the peptides into the membrane plane is also found to be an important characteristic for the fusion process. A shallow penetration, extending only to the central aliphatic core region, is likely responsible for the destabilization of the lipids required for coalescence of the apposing membranes and fusion. PMID- 11426691 TI - Viral fusion peptides: a tool set to disrupt and connect biological membranes. AB - The structure and function of viral fusion peptides are reviewed. The fusion peptides of influenza virus hemagglutinin and human immunodeficiency virus are used as paradigms. Fusion peptides associated with lipid bilayers are conformationally polymorphic. Current evidence suggests that the fusion-promoting state is the obliquely inserted alpha-helix. Fusion peptides also have a tendency to self-associate into beta-sheets at membrane surfaces. Although the conformational conversion between alpha- and beta-states is reversible under controlled conditions, its physiological relevance is not yet known. The energetics of peptide insertion and self-association could be measured recently using more soluble "second generation" fusion peptides. Fusion peptides have been reported to change membrane curvature and the state of hydration of membrane surfaces. The combined results are built into a model for the mechanism by which fusion peptides are proposed to assist in biological membrane fusion. PMID- 11426692 TI - Hydrophobic-at-interface regions in viral fusion protein ectodomains. AB - In this chapter we shall describe how to apply the hydrophobicity-at-interface scale, as proposed by Wimley and White [Wimley, W. C. and White, S. H. (1996) Nature Struct. Biol. 3:842-848], to the detection of amino acid sequences of viral envelope glycoproteins putatively engaged in interactions with the target membranes. In addition, a new approach will be briefly introduced to infer the bilayer location at equilibrium of membrane-partitioning sequences. The use of these new procedures may be important in describing the molecular mechanism leading to the formation of a fusion pore by viral glycoproteins. PMID- 11426693 TI - Functional domains within fusion proteins: prospectives for development of peptide inhibitors of viral cell fusion. AB - The entry of enveloped viruses into host cells is accomplished by fusion of the viral envelope and target plasma membrane and is mediated by fusion proteins. Recently, several functional domains within fusion proteins from different viral families were identified. Some are directly involved in conformational changes after receptor binding, as suggested by the recent release of crystallographically determined structures of a highly stable core structure of the fusion proteins in the absence of membranes. However, in the presence of membranes, this core binds strongly to the membrane's surface and dissociates therein. Other regions, besides the N-terminal fusion peptide, which include the core region and an internal fusion peptide in paramyxoviruses, are directly involved in the actual membrane fusion event, suggesting an "umbrella" like model for the membrane induced conformational change of fusion proteins. Peptides resembling these regions have been shown to have specific antiviral activity, presumably because they interfere with the corresponding domains within the viruses. Overall, these studies shed light into the molecular mechanism of membrane fusion induced by envelope glycoproteins and suggest that fusion proteins from different viral families share common structural and functional motifs. PMID- 11426694 TI - Insights into a structure-based mechanism of viral membrane fusion. AB - A number of different viral spike proteins, responsible for membrane fusion, show striking similarities in their core structures. The prospect of developing a general structure-based mechanism seems plausible in light of these newly determined structures. Influenza hemagglutinin (HA) is the best-studied fusion machine, whose action has previously been described by a hypothetical "spring loaded" model. This model has recently been extended to explain the mechanism of other systems, such as HIV gp120-gp41. However, evidence supporting this idea is insufficient, requiring re-examination of the mechanism of HA-induced membrane fusion. Recent experiments with a shortened construct of HA, which is able to induce lipid mixing, have provided evidence for an alternative scenario for HA induced membrane fusion and perhaps that of other viral systems. PMID- 11426695 TI - The role of the transmembrane and of the intraviral domain of glycoproteins in membrane fusion of enveloped viruses. AB - Fusion of enveloped viruses with their target membrane is mediated by viral integral glycoproteins. A conformational change of their ectodomain triggers membrane fusion. Several studies suggest that an extended, triple-stranded rod shaped alpha-helical coiled coil resembles a common structural and functional motif of the ectodomain of fusion proteins. From that, it is believed that essential features of the fusion process are conserved among the various enveloped viruses. However, this has not been established so far for the highly conserved transmembrane and intraviral sequences of fusion proteins. The article will focus on the role of both sequences in the fusion process. Recent studies from various enveloped viruses strongly imply that a transmembrane domain with a minimum length is required for later steps of membrane fusion, i.e., the formation and enlargement of the aqueous fusion pore. Although no specific sequence of the TM is necessary for pore formation, distinct properties and motifs of the domain may be obligatory to ascertain full fusion activity. However, with some exceptions, the intraviral domain seems to be not required for fusion activity of viral fusion proteins. PMID- 11426696 TI - Virus membrane fusion proteins: biological machines that undergo a metamorphosis. AB - Fusion proteins from a group of widely disparate viruses, including the paramyxovirus F protein, the HIV and SIV gp160 proteins, the retroviral Env protein, the Ebola virus Gp, and the influenza virus haemagglutinin, share a number of common features. All contain multiple glycosylation sites, and must be trimeric and undergo proteolytic cleavage to be fusogenically active. Subsequent to proteolytic cleavage, the subunit containing the transmembrane domain in each case has an extremely hydrophobic region, termed the fusion peptide, or at near its newly generated N-terminus. In addition, all of these viral fusion proteins have 4-3 heptad repeat sequences near both the fusion peptide and the transmembrane domain. These regions have been demonstrated from a tight complex, in which the N-terminal heptad repeat forms a trimeric-coiled coil, with the C terminal heptad repeat forming helical regions that buttress the coiled-coil in an anti-parallel manner. The significance of each of these structural elements in the processing and function of these viral fusion proteins is discussed. PMID- 11426697 TI - On the mechanism of intracellular membrane fusion: in search of the genuine fusion factor. AB - Intracellular membrane fusion events require a general protein machinery that functions in vesicular traffic and in assembly and maintenance of organelles. An array of cytosolic and integral membrane proteins are currently identified, and in conjunction with ongoing detailed structural studies, rapid progress is made in understanding basic features of the overall mechanism of the fusion machinery, but above all a proper appreciation of its enormous complexity. Thus a highly sophisticated level of regulation of the different steps involved in tethering, docking and merging itself is apparent. Apart from the relevance of protein protein interactions, also a role of distinct lipids is gradually emerging, particularly in fusion. However, although various suggestions have been made recently, largely based upon in vitro studies, the identity of the actual fusion factor(s) remains to be determined. PMID- 11426698 TI - Factors influencing the stability of alpha-helices and beta-strands in thermophilic ribonuclease H. AB - Understanding the influence of structural parameters is crucial to enhance the thermal stability of proteins. In this work, the stability (deltaG) of residues in different secondary structures of Ribonuclease H (RNase H) has been analyzed with 48 amino acid properties. The properties reflecting hydrophobicity show a good correlation with stability. Further, the linear distribution of surrounding hydrophobicity in alpha-helices, obtained from the three dimensional structure of thermophilic RNase H, agrees well with experimental deltaG values. Moreover, the stability parameters correlate better in alpha-helices than those did in beta strand segments. Multiple regression analysis, incorporating combinations of three properties from among all possible combinations of the 48 properties, increased the correlation coefficient to 0.77. PMID- 11426699 TI - Enzymatic preparation of heparin disaccharides as building blocks in glycosaminoglycan synthesis. AB - Pharmaceutical heparin and heparan sulfate, isolated from a side-stream of a commercial heparin manufacturing process, have been enzymatically depolymerzed with heparin lyases obtained from Flavobacterium heparinun. Heparin afforded a trisulfated disaccharide product that was recovered from the reaction mixture using gel permeation chromatography. Heparan sulfate afforded unsulfated disaccharide that was conveniently recovered from the product mixture by ion exchange chromatography. Both disaccharides were obtained in gram amounts at 90% or higher purity. Both enzymatically prepared disaccharides were chemically protected to prepare building blocks required for the future chemical synthesis of therapeutically valuable heparin oligosaccharides. PMID- 11426700 TI - Purification of lactoperoxidase from bovine milk and investigation of the kinetic properties. AB - Lactoperoxidase (LPO) was purified from bovine milk using Amberlite CG 50 H+ resin, CM Sephadex C-50 ion-exchange chromatography, and Sephadex G-100 gel filtration chromatography. During the purification steps, the activity of enzyme was measured using 2,2'-azino-bis (3-ethylbenzthiazoline-6 sulfonic acid) diamonium salt (ABTS) as a chromogenic substrate at pH 6. Optimum pH and optimum temperature values for LPO were determined for ABTS, p-phenylendiamine, catechol, epinephrine, and pyrogallol as substrates, and then Km and Vmax values for the same substrate were obtained by means of Lineweaver-Burk graphics. The purification degree of the enzyme was controlled by SDS-PAGE and Rz (A412/A280) values. Km values, at optimum pH and 20 degrees C, were 0.197 mM, 0.063 mM, 0.64 mM, 25.2 mM, and 63.95 mM for p-phenylendiamine, ABTS, epinephrine, pyrogallol, and catechol, respectively. Vmax values, at optimum pH and 20 degrees C, were 3.5x10(-5) EU/mL, 4.0x10(-5) EU/mL, 5.8x10(-4) EU/mL, 8.4x10(-4) EU/mL, and 1.01x10(-3) EU/mL for the same substrates, respectively. p-Phenylendiamine was first found as a new substrate for LPO. PMID- 11426701 TI - Purification and identification of N-glycolylneuraminic acid (Neu5Gc) from the holothuroidea Gumi, Cucumaria echinata. AB - N-Glycolylneuraminic acid (Neu5Gc), precious sialic acid which could not be synthesized by a chemical method, occurrs in the body of holothuroidea, Gumi Cucumaria echinata. Gumi contains 85% of total sialic acid, as Neu5Gc, in the body. Neu5Gc was purified from dry powder of the body using Dowex 1-x8 (HCOO* form) anion exchange chromatography after mild acid hydrolysis with 0.1 N trifluoroacetic acid. Using GC-MS and 1H-NMR spectroscopy, the purified Neu5Gc was correctly identified to be Neu5Gc. The purity of Neu5Gc was more than 99%. This is the first report of purification and identification of Neu5Gc from holothuroidea by using anion exchange chromatography, GC-MS, and 1H-NMR. PMID- 11426702 TI - Suitability of animals' purified milk caseins and their subunit kappa-caseins as substrates for subtilisin and trypsin. AB - Acid casein and kappa-casein were purified from different species of animal's milk, such as cow, sheep, goat, and water buffalo. These caseins were used as substrates for commercially available subtilisin and trypsin. It was established that, when acid caseins were used as a substrate for subtilisin, cow acid casein was found to be a better substrate for the enzymes, compared to other animals' milk casein. It was suggested that this acid casein has significantly more aromatic amino acids, as compared to arginine and lysine. K(M) and Vmax values, which were obtained for cow kappa-casein, showed that cow kappa-casein was a better susbstrate for trypsin than the others, suggesting that cow kappa-casein has a rich content of lysine, arginine, and aromatic amino acids by comparison with the others. The calculated C/N ratio also supports this suggestion. PMID- 11426703 TI - Isolation of total RNA from pollens. AB - Isolation of total RNA from plant materials has been difficult, due to the presence of complex organic substances and the associated pigmentation. In fact, there is a dearth of standardized protocols for isolating total RNA from pollens. To find a simple and reliable method for isolating total RNA from pollen, four methods, viz. phenol/SDS (PS), guanidine HCl (GH), tri-reagent (TR), and modified SDS-betaME (SB) were tested with fresh pollen of Ricinus communis (procured at 70 degrees C) and pollen dried at 30-37 degrees C. The quality and quantity of RNA was superior for the material processed at -70 degrees C. SB gave the highest RNA yield (2.35 mg/g, OD260/280 >2.0), compared to other methods. The results obtained by the SB method were found to be comparable with the widely used tri reagent method. This was validated with other pollens of Imperata cylindrica and Xanthium strumarium. The yield obtained from graded amounts of pollen was consistent with SB, compared to the TR method. The RNA isolated by SB gave good quality mRNA for synthesizing cDNA. The SDS-betaME method is simple, efficient, and uses less expensive reagents. Hence, we recommend the modified SDS-betaME method for isolating total RNA from pollens. PMID- 11426704 TI - Distribution of amino acid residues and residue-residue contacts in molecular chaperones. AB - The amino acid distribution and residue-residue contacts in molecular chaperones are different when compared to normal globular proteins. The study of molecular chaperones reveals a different surrounding environment to exist for the residues Cys, Trp, and His which may play an important role in determining the chaperone structures. Unlike globular proteins, it has been observed that a one-to-one correspondence between the amino acid distribution in a sequence and the structures of molecular chaperones. The preference of amino acid residues surrounding all 20 types of residues in secondary structures and their accessible surface areas have been analysed. PMID- 11426705 TI - Mapping of hiv-1 Gag epitopes recognized by polyclonal antibodies using gene fragment phage display system. AB - Phage display has emerged as a powerful technique for mapping epitopes recognised by monoclonal and polyclonal antibodies. We have recently developed a simple gene fragment phage display system and have shown its utility in mapping epitope recognised by a monoclonal antibody. In the present study, we have employed this system in mapping epitopes recognised by polyclonal antibodies raised against HIV 1 capsid protein, p24 which is derived from proteolytic cleavage of Gag polyprotein. HIV-1 gag DNA was fragmented by DNase I and the fragments (50-250 bp) were cloned into gene-fragment phage display vector to construct a library of phages displaying peptides. This phage library was used for affinity selection of phages displaying epitopes recognised by rabbit anti-p24 polyclonal antibodies. Selected phages contained sequences from two discrete regions of p24, demonstrating the presence of two antigenic regions. The DNA sequences encoding these regions were also cloned and expressed as GST fusion proteins. The immunoreactivity of these epitopes as GST fusion proteins, or as phage-displayed peptides, was comparable in ELISA system using same anti-p24 polyclonal antibodies. The results indicate that the gene-fragment based phage display system can be used efficiently to identify epitopes recognised by polyclonal antibodies, and phage displayed epitopes can be directly employed in ELISA to detect antibodies. PMID- 11426706 TI - WinPep 2.11: novel software for PC-based analyses of amino acid sequences. AB - WinPep 2.11 is the latest version of a user-friendly, versatile software for the analysis of amino acid sequences. WinPep was developed for Windows 95/98 and WindowsNT. Because of the standard windows techniques (copy & paste) and an intuitive user interface, no lengthy training is required to work with this program. Main features include calculation of molecular weight, isoelectric point, and molar absorption coefficients, as well as simulation of sequence specific cleavage. In addition to data analysis, data presentation is greatly assisted by the program, e.g., by producing helical wheel displays and hydropathy plots. The implemented functions of WinPep and it's usage are described in this publication. PMID- 11426707 TI - Rapid fractionation of bovine transferrin using immobilized gangliosides. AB - Bovine transferrin (BTF) was fractionated from bovine whey using ganglioside affinity chromatography. After loading the immobilized matrix with a 2% whey solution, the matrix was washed with sodium acetate buffer at pH 4 containing 1 M NaCl before elution of BTF with sodium phosphate buffers at pH 7. Concanavalin-A affinity and ion exchange chromatography were used for further purification. The ganglioside column showed a 74.2% BTF recovery from whey and BTF was enriched to 61% purity with ion exchange chromatography. Bovine transferrin was identified by SDS-PAGE and western analysis. The Concanavalin-A affinity and ion exchange chromatography steps enriched BTF in the samples and removed other whey proteins from ganglioside purified fractions. These results indicate that immobilized ganglioside can be used to fractionate BTF from bovine whey. Our novel ganglioside affinity chromatography is rapid and efficient for the fractionation of BTF from whey. PMID- 11426708 TI - Penetration of erythrocytes by merozoites of mammalian and avian malarial parasites. 1969. PMID- 11426709 TI - The fate of merozoites. PMID- 11426710 TI - Integrative neuroimmunomodulation of gastrointestinal function during enteric parasitism. AB - Enteric helminths have a significant impact on the structure, function, and neural control of the gastrointestinal (GI) tract of the host. Interactions between the host's nervous and immune systems redirect activity in neuronal circuits intrinsic to the gut into an alternative repertoire of defensive and adaptive motor programs. Gut inflammation and activation of the enteric neuroimmune axis play integral roles in the dynamic interaction between host and parasite that occurs at the mucosal surface. Three inter-related themes are stressed in this review to underscore the pivotal role that neural control mechanisms play in the host's GI tract functional responses to enteric parasitism. First, we address the discovery that signaling molecules of both parasite and host origin can reorient the dynamic ecology of enteric host parasite interactions. Second, we explore what has been learned from investigations of altered gut propulsive and secretomotor reflex activities that occur during enteric parasitic infections and the emerging picture derived from these studies that elucidates how nerves help facilitate and orchestrate functional reorganization of the parasitized gut. Third, we provide an overview of the direct impact that enteric parasitism has on nerve cell function and neurotransmission pathways in both the enteric and central nervous systems of the host. In summary, this review highlights and clarifies the complex mechanisms underlying integrative neuroimmunophysiological responses to the presence of both invasive and noninvasive enteric helminths and identifies directions for future research investigations in this highly important but understudied area. PMID- 11426711 TI - Photo- and geo-orientation by Echinostome cercariae results in habitat selection. AB - The effect of light and gravity on orientation was studied in cercariae of 4 echinostome species: Pseudechinoparyphium echinatum, Echinostoma revolutum, Hypoderaeum conoideum, and Isthmiophora melis. The cercariae were placed into vertical and horizontal cuvettes, illuminated with 2 different light intensities from various directions, and their distribution recorded for 6 hr Each species showed its individual pattern of horizontal photo-orientation and geo orientation, with distinct changes during the time after emerging. The geo orientation was controlled differently in each species by the intensity and the direction of light radiation. The different orientation patterns suggest functions such as leaving the habitats of the host-snails emitting the cercariae, dispersal, and frequenting the microhabitats of potential hosts. The high diversity of orientation patterns among the species that originated from the same first intermediate host Lymnaea stagnalis in the same ponds and that invade similar host spectra suggests adaptations to different ecological conditions. PMID- 11426712 TI - Human cerebral cysticercosis: immunolocalization of a sodium-dependent glucose cotransporter (SGLT) in larval and adult tapeworms. AB - Light microscopic immunocytochemistry was used to examine human brain cysticerci resected from the fourth ventricles of patients who had not been treated with anthelminthic drugs. Tissues were examined from 3 different patients undergoing surgery for treatment of hydrocephalus. A rabbit polyclonal antiserum to the peptide corresponding to amino acids 564-575 unique to the rabbit sodium dependent, SGLT1 glucose cotransporter labeled with immunoperoxidase, localized immunoreactive SGLT epitopes. This antibody localizes SGLT1 in the apical brush borders of human enterocytes, but is negative in cytoplasm, as well as lateral and basal enterocyte membranes. Taenia solium neurocysticerci were SGLT positive; transporter protein was highly expressed on the surface microvilli of the external cyst wall. The well-developed network of small and larger osmoregulatory ducts within racemose larval cystcerci displayed high expression of SGLT cotransporter, consistent with a resorptive function for this system of tubules. Because water is cotransported with glucose molecules by the SGLT protein, its high expression in neurocysticerci may contribute to the expansive growth of these larvae in subarachnoid and intraventricular sites. The SGLT epitopes were also immunolocalized in gravid proglottids of Taenia saginata, indicating that cotransporter expression persisted in intestinal-dwelling, adult tapeworms. Cotransporter antibody was abundantly localized at the proglottid tegumentary surface and in the lateral osmoregulatory ducts, analogous to the SGLT localization in cysticerci. Furthermore, high expression of this cotransporter was seen in the branches of the uterus, suggesting that SGLT-mediated absorption of glucose and water has an important functional role within the reproductive system of adult tapeworms. PMID- 11426713 TI - Relaxing and contracting activities of heartworm extract on isolated canine abdominal aorta. AB - Effect of adult heartworm (HW) crude extract on isolated canine abdominal aortic strips precontracted with noradrenaline was examined by recording isometric changes in tension. HW extract caused contraction of the aortic strip at a low concentration (LC) and its relaxation at a high concentration (HC). In aortic strips without endothelium, LC extract elicited a contraction similar to that in the strips with endothelium, whereas HC extract failed to produce any relaxation but instead produced a contraction. The relaxing effect of HC extract was blocked after treatment with 300 microM NG-nitro-L-arginine methyl ester hydrochloride, with reversal by additional treatment with 3 mM L-arginine. It was also markedly reduced or abolished after treatment with 3 microM oxyhemoglobin or 1 microM methylene blue. Fractionation of HW extract by high-performance liquid chromatography revealed that the relaxing and contracting activities are due to different substances in the extract. The results indicate that HW extract contains 2 different vasoactive substances, 1 causing contraction of canine abdominal aorta via a direct action on the smooth muscle, and the other its relaxation indirectly by releasing nitric oxide from endothelial cells. These vasoactive substances might play a role in HW extract-induced shock in dogs, and in the pathogenesis of HW infection. PMID- 11426714 TI - Umingmakstrongylus pallikuukensis (Nematoda: Protostrongylidae) in gastropods: larval morphology, morphometrics, and development rates. AB - Morphological and morphometric aspects of larval development of Umingmakstrongylus pallikuukensis in Deroceras laeve and the effects of temperature on development rates in D. laeve and Deroceras reticulatum were investigated in the laboratory. Larval stages were best differentiated by separation of cuticular sheaths, tail structure, and viability following digestion. Growth in body and esophagus width was observed during the first-stage within the intermediate host, but the major increases in body length and width occurred immediately following the second molt. Larval development in D. laeve and D. reticulatum occurred more rapidly at warmer temperatures. The calculated threshold temperatures were 8.5 and 9.5 C in D. laeve and D. reticulatum, respectively, and 167 degree-days were required for development to third-stage larvae (L3) in both hosts. These thresholds are higher than those calculated from published data for the closely related Muellerius capillaris (4.2 C) but are similar to those for the more distantly related northern protostrongylid, Elaphostrongylus rangiferi (8.3-10.3 C). Conversely, degree-days required for development to infective L3 were more similar among the Muelleriinae than between this group and the Elaphostrongylinae. Developmental parameters for protostrongylid larvae may be influenced both by the environment and by features of the parasites and the intermediate hosts, including phylogeny. PMID- 11426715 TI - Corynosoma cetaceum in the stomach of franciscanas, Pontoporia blainvillei (Cetacea): an exceptional case of habitat selection by an acanthocephalan. AB - Adult acanthocephalans are typically found in the intestine of vertebrates, where they can readily absorb nutrients. However, Corynosoma cetaceum has been frequently reported in the stomach of cetaceans from the Southern Hemisphere. The ecological significance of this habitat was investigated by examining data on number, sex ratio, maturity status, biomass, and fecundity of C. cetaceum in different parts of the digestive tract of 44 franciscanas Pontoporia blainvillei. Individual C. cetaceum occurred in the pyloric stomach (PS) and, to lesser degrees, in the duodenal ampulla (DA) and the main stomach (MS). Females outnumbered males in all chambers, although the sex ratio was closer to 1:1 in the MS; there also was a higher proportion of nongravid females, with a smaller biomass in the MS than in the PS and the DA. This evidence suggests that cystacanths are released from prey tissues in the MS, where entire prey are reduced to semi-fluid chyme. The 3 chambers harbored gravid females that did not differ significantly in mean biomass or fecundity. The maturity status of females was nearly identical between the PS and the DA. In the MS, the higher proportion of non-gravid females is probably due to the occurrence of newly recruited females to this site. Mean biomass and fecundity of gravid females covaried strongly and positively among chambers within hosts. These results suggest that there are no major differences between the 3 chambers with respect to the suitability for reproduction by C. cetaceum. However, although the MS is the largest chamber, it harbored the smallest number of gravid females. Interestingly, worms were largely restricted to the aboral portion of the MS, a sheltered region where a concentration of chyme, and thus nutrient availability, likely occurs. Linear distribution differences of gravid female C. cetaceum at increasing intensities suggest that reproductive females occupy chambers according to available space. In summary, the stomach should be considered the main habitat for C. cetaceum. The choice of this habitat is puzzling because other Corynosoma species occur in the intestine, and because the stomach of cetaceans is not an absorptive site. PMID- 11426716 TI - Fecundity of Hoplopleura scapteromydis (Phthiraptera: Anoplura, Hoplopleuridae), ectoparasite of Scapteromys aquaticus (Rodentia: Muridae) in Rio de La Plata marshlands, Argentina. AB - Density and age structure of lice collected from captured Scapteromys aquaticus rodents were studied to estimate the fecundity of Hoplopleura scapteromydis. The number of eggs with a visible embryo inside (E), nymphs (N), adult males (AM), and adult females (AF) were recorded for each rodent. For the ith rodent, the fecundity of H. scapteromydis, F(i), was estimated as F(i) = [[E(i) + N(i)]/2]/AF(i)/T, where T represents the period of preimaginal development (unknown and arbitrarily considered as T = 1), and the sex ratio of the preimaginal stages was supposed to be 1:1. In order to look for density-dependent effects, F(i) was plotted against AM(i), this being an independent estimation of infrapopulation density. The number of rodents suitable for AF and AM calculations was 38 (57% of the parasitized animals). Almost 95% had a low-to moderate louse burden (1 < AM < 30) and were captured every season, whereas only 3% had a heavy (61 < AM < 70, captured in winter) or very heavy (AM > 80, captured in summer) louse burden. The extreme values of F were 0.63 and 18 (1.3 and 36 if both sexes were considered). High-to-moderate F-values (F > 5) were estimated in only 5 rodents that exhibited low louse density, whereas low F values (F < 5) were found at all louse densities. Notwithstanding the tendency toward an inverse relationship between fecundity and infrapopulation density, the correlation was not significantly different from 0. PMID- 11426717 TI - Seasonal studies on commensal rats and their ectoparasites in a rural area of Egypt: the relationship of ectoparasites to the species, locality, and relative abundance of the host. AB - The present study was carried out in 3 villages, namely Kafr Ayoub Soliman, Kafr Ibrahim El-Aidi, and El-Sa'adat, Sharqiya Governorate, Egypt. A total of 519 rats was collected from the 3 study sites: 46.6% Rattus rattus, and 53.4% Rattus norvegicus. A total of 20,643 ectoparasites was recovered from R. rattus: 33.3% mites, 33.8% fleas, and 32.9% lice. From R. norvegicus a total of 40,997 ectoparasites was recovered: 28.9% mites, 31% fleas, and 40.1% lice. Three common mite species were recovered from both rat hosts, i.e., Ornithonyssus bacoti, Radfordia ensifera, and Laelaps nuttalli. Three common flea species were also recovered from both rat hosts, i.e., Echidnophaga gallinacea, Leptopsylla segnis, and Xenopsylla cheopis. Polyplax spinulosa was the only dominant louse species that infested both rat hosts. Rats did not show a definite breeding season, and the seasonal rat indices were generally low in different study sites. There were no significant differences between the prevalence of each of mites, fleas, and lice in both rat species. The total general indices of mites and fleas, on the other hand, was significantly higher in R. norvegicus. The general index of X. cheopis was high and ranged between 5.9 in R. rattus and 14.5 in R. norvegicus. Season-related changes were observed in the general index of each of L. segnis infesting both rat species and R. ensifera and O. bacoti infesting R. norvegicus. The prevalence and general indices of some ectoparasites showed differences related to the locality of their rat hosts. Seasonal changes in the general indices of some ectoparasites paralleled seasonal changes in the relative abundance of their rat hosts. PMID- 11426718 TI - Lipids of three microsporidian species and multivariate analysis of the host parasite relationship. AB - Sporal lipids of 3 microsporidia, Encephalitozoon cuniculi from mammals and Glugea atherinae and Spraguea lophii from fishes, were investigated. High phospholipid levels were found (54.8-64.5% of total lipids), which is in agreement with the presence of highly developed internal membranes in microsporidian spores. Sphingomyelin was not detected in G. atherinae. Triglycerides (less than 10% of total lipids), cholesterol, and free fatty acids were identified in all species. Analysis of fatty acids from the phospholipid fraction revealed the predominance of docosahexaenoic acid (30-40% of total phospholipid fatty acids) in G. atherinae and S. lophii and oleic acid (25.8% of total phospholipid fatty acids) in E. cuniculi. The 3 microsporidia possessed a significant amount of branched-chain fatty acids (iso and anteiso forms) not found in the hosts, supporting the existence of some parasite-specific metabolic steps for these fatty acids. On the basis of phospholipid fatty acid profiles, host-parasite relationships were investigated through correspondence factorial analysis. It shows 3 distinct clusters with the first corresponding to fishes, the second to fish parasites, and the third to E. cuniculi and its host cell. These data suggest that the mammal microsporidia developing within parasitophorous vacuoles are more dependent on host cells than the fish microsporidia that induce cystlike structures. PMID- 11426719 TI - Frontal filament morphogenesis in the salmon louse Lepeophtheirus salmonis. AB - The objective of the present study was to understand how and when the frontal filament (FF) in the salmon louse Lepeophtheirus salmonis is produced by examining the sequence of morphological changes leading to FF production in the copepodid and early chalimus stages. Atlantic salmon (Salmo salar) were heavily infested with newly molted copepodids. Sea lice were sampled prior to infestation and at 1, 2, 3, 4, 5, 6, 7, 8, and 9 days postinfestation. FF morphogenesis from newly molted copepodid to chalimus II, i.e., through 2 molts, was studied using high-resolution light microscopy of serial transverse and sagittal resin sections. Three groups of cells, identified as A, B, and C, are thought to be involved in the production of the secretions (S1 and S2) that form the filament material. The amount and shape of S1 and S2 and their association with B- and C group cells, respectively, changed with the molt cycle. The following scenario for FF development is proposed: the first secretion to form after the molt for both copepodid and chalimus stages is S1, and it is formed by B-group cells and becomes the basal plate of the external FE C-group cells produce S2 during mid intermolt to premolt stage. The S2 becomes the stem of the external FE In premolt larvae, S1 and S2 were contained within a cuticle-lined invagination that had a form similar to that of the extruded filament. The axial duct present in both copepodid and chalimus originates from the A-group cells and probably carries a secretion used to attach the filament to the host. This study provides strong evidence that L. salmonis produces a new filament with each molt, creating the possibility of using a sea lice control method based on interference with filament production more feasible. PMID- 11426720 TI - Chemical and physical factors affecting the excystation of Cryptosporidium parvum oocysts. AB - Cryptosporidium parvum oocysts were examined to ascertain excystation requirements and the effects of gamma irradiation. Oocysts and excysted sporozoites were examined for dye permeability and infectivity. Maximum excystation occurred when oocysts were pretreated with acid and incubated with bile salts, and potassium or sodium bicarbonate. Pretreatment with Hanks' balanced salt solution or NaCl lowered excystation; however, this effect was overcome with acid. Sodium ions were replaceable with potassium ions, and sodium bicarbonate was replaceable with sodium phosphate. Oocysts that received 200 krad irradiation excysted at the same rates as nonirradiated oocysts (95%), the excystation rates were lowered (50%) by 2,000 krad, and no excystation was observed by 5,000 krad. No differences were observed between the propidium iodide (PI) permeability of untreated oocysts and oocysts treated with 200 krad, while 92% of oocysts were PI positive after 2,000 krad. Most of the sporozoites exposed to 2,000 krad were not viable as indicated by the dye permeability assay. The oocysts irradiated with 200 and 2,000 krad infected cells, but no replication was observed. The results suggest that gamma-irradiated oocysts may still be capable of excystation and apparent infection; however, because the sporozoites could not reproduce they must not have been viable. PMID- 11426721 TI - Inhibitory role of antibodies in the development of Taenia solium and Taenia crassiceps toward reproductive and pathogenic stages. AB - Untreated Taenia solium cysticerci obtained from different naturally infected pigs vary notably in their capacity to develop into intestinal tapeworms in prednisolone-treated hamsters, whereas cells derived from Taenia crassiceps cysticerci after 2 mo of infection almost always develop to cysticerci in the peritoneal cavity of susceptible BALB/cAnN mice. Preincubation of whole cysticerci or parasite cells with mice immunoglobulins raised against an 18-mer peptide epitope (GK-1) common to both parasites significantly interferes with both transformations. These crippling effects of antiparasite antibodies suggest new forms of immunological interference with parasite biology other than simple killing. Antibodies that cripple biological functions of the parasite, e.g., their development to reproductive or pathogenic stages, make them important protagonists in taeniasis/cysticercosis disease as classic parasitocidal antibodies. Different serum levels of crippling antibodies in the infected pigs could be responsible for the varied ability of cysticerci to convert to tapeworms. Antigens capable of inducing crippling antibodies, e.g., GK-1, could be useful as a therapeutic vaccine for pigs in order to reduce parasite transmission. PMID- 11426722 TI - Cellular immune response and Th1/Th2 cytokines in human neurocysticercosis: lack of immune suppression. AB - Some reports have suggested that human neurocysticercosis (NCC) induces immunosuppression. To test this hypothesis, we performed a study on active cases of NCC who had not received cestocidal or immunosuppressive treatments. We examined blood counts and specific T cell markers (CD3, CD4, and CD8) by flow cytometry and found no differences between patients with NCC and healthy individuals. Both groups responded to concanavalin A (Con A), and patients with NCC responded more to a parasite crude antigen than uninfected individuals. Peripheral blood mononuclear cells were examined for interleukin (IL)-2, interferon-gamma, IL-10, and IL-4 mRNA. Regardless of infection status, more than 60% of individuals synthesized IL-2 mRNA and, less frequently, the other cytokines. These data suggest that immunosuppression does not occur in NCC patients. PMID- 11426723 TI - Kinetics and characterization of cellular responses in the peritoneal cavity of mice infected with Taenia crassiceps. AB - Changes in the leukocyte population of the peritoneal cavity ensue immediately after infection with Taenia crassiceps metacestodes. Basophils and neutrophils decrease, whereas macrophages, monocytes, and lymphocytes increase to reach only modest levels by 6 wk and then diminish to nearly disappear by 15 wk when the parasite begins rapid reproduction. Eosinophils also appear early in infection, but then abate to lower levels that persist. In late infections, when the mass of cysticerci equals that of the mouse, the cysticerci grow among surprisingly few inflammatory cells. Mingling with the peritoneal inflammatory cells is a number of odd-looking cells that could correspond to the metaplasic mesothelial cells of the host or be of parasite origin. These cells are multinucleated, they aggregate in varigerated clusters, and form cystic structures in vitro; they also bind specific anti-T. crassiceps antibodies and specific T. crassiceps DNA probes in their nuclei. When the peritoneal cell exudate is reinjected intraperitoneally into naive mice, the odd-looking cells subsist for months, inducing in the host the synthesis of specific anti-T. crassiceps antibodies and immune resistance to challenge but do not reassemble into cysticerci even after 6 mo of inoculation. The early appearance and the immunogenic and antigenic properties of these odd looking cells suggest they are important protagonists in the early host-parasite confrontation when the outcome of infection is set. PMID- 11426724 TI - The response of Gregarina niphandrodes (Apicomplexa: Eugregarinida: Septatina) to host starvation in Tenebrio molitor (Coleoptera: Tenebrionidae) adults. AB - Numerous studies of host starvation have emphasized pathological effects of parasites on their insect host, but little attention has been focused on the effects of host starvation on the parasites. This study addressed the possibility that parasite life-cycle events could be manipulated by withholding food from the host. The system used was Gregarina niphandrodes (Apicomplexa: Eugregarinida) in Tenebrio molitor (Coleoptera: Tenebrionidae) adults. Gregarine gametocyst formation and shedding ceased after 1 day in starved beetles but continued in fed controls. There were no statistically significant differences between total lengths of associated (3 of 5 trials) or unassociated (5 of 5 trials) gregarines found between experimental and control groups, but average numbers of the 2 life cycle events were generally higher in fed hosts than in starved ones. If infected, fed control beetles continued to form gametocysts throughout the 7-day trial periods, and gametocysts could be observed in the gut. Starved experimental beetles had no gametocysts in their guts. Refeeding of starved beetles after 4 days resulted in resumption of gametocyst formation and shedding. The studies demonstrated that the gregarine life cycle could be stopped and then started at the gametocyst formation stage like an off/on switch, simply by withholding food from, then refeeding, the host. PMID- 11426725 TI - Transmission of hepatozoon canis to dogs by naturally-fed or percutaneously injected Rhipicephalus sanguineus ticks. AB - Hepatozoon canis is an apicomplexan protozoan parasite of dogs, prevalent in Asia, Africa, and southern Europe. Experimental transmission of H. canis to dogs was performed with laboratory-reared Rhipicephalus sanguineus nymphs that fed on a naturally infected dog or were percutaneously injected with canine blood containing H. canis gamonts. Dogs were inoculated by oral ingestion of adult ticks containing H. canis oocysts. Transstadial transmission of H. canis was recorded, whereas transovarial transmission could not be demonstrated. Oocysts were detected in 85% of the adult ticks that had engorged as nymphs on an infected dog and in 61% of the adult ticks resulting from nymphs injected percutaneously with blood from the same dog. Nine of 12 dogs (75%) inoculated with naturally fed or percutaneously injected ticks became parasitologically positive, and all showed seroconversion. Meronts were initially detected in the bone marrow 13 days postinoculation and gamonts 28 days after infection. The variation in the time of initial detection of parasitemia among infected dogs and the rapid appearance of gamonts in dogs immunosuppressed with corticosteroids suggest that immune mechanisms play an important role in controlling H. canis parasitism. The artificial acquisition of Hepatozoon parasites by percutaneous injection of ticks, demonstrated here for the first time, may serve as a useful tool for studies on transmission, vector-host relationships, and the immunology of infection with Hepatozoon species. PMID- 11426726 TI - First isolation of Neospora caninum from the feces of a naturally infected dog. AB - Neospora caninum is a major cause of abortion in cattle worldwide. Cattle become infected with N. caninum by ingesting oocysts from the environment or transplacentally from dam to fetus. Experimentally, dogs can act as definitive hosts, but dogs excrete few oocysts after ingesting tissue cysts. A natural definitive host was unknown until now. In the present study, N. caninum was isolated from the feces of a dog. Gerbils (Meriones unguiculatus) fed feces from the dog developed antibodies to N. caninum in the Neospora caninum agglutination test, and tissue cysts were found in their brains. Neospora caninum was isolated in cell culture and in gamma-interferon gene knockout mice inoculated with brain homogenates of infected gerbils. The DNA obtained from fecal oocysts of the dog, from the brains of gerbils fed dog feces, and from organisms isolated in cell cultures inoculated with gerbil brains was confirmed as N. caninum. The identification of N. caninum oocyst by bioassay and polymerase chain reaction demonstrates that the dog is a natural definitive host for N. caninum. PMID- 11426727 TI - Identification of a collagen-binding protein from Necator americanus by using a cDNA-expression phage display library. AB - A phage display library was made starting from a cDNA library from the hematophagous human parasite Necator americanus. The cDNA library was transferred by polymerase chain reaction (PCR) cloning into phage display vectors (phagemids), using specially designed primers such that proteins would be expressed as fusions with the C-terminal part of the phage coat protein pVI. The vectors used are multicloning site variants of the original pDONG vectors described by Jespers et al. (1995). Electroporation of the ligation mixtures into electrocompetent Escherichia coli TGI cells yielded 3 x 10(8) pG6A, 1.9 x 10(8) pG6B, and 1 x 10(8) pG6C transfectants for N. americanus. The final libraries consisted of a mix of equal numbers of insert-containing phages from the A, B, and C libraries. Selection of phages for binding to human collagen was performed. Four rounds of panning on human collagens I and III resulted in a significant enrichment of collagen-binding phages from the N. americanus libraries. PCR analysis revealed various insert lengths; however, sequence determination indicated that all phages contained the same protein, albeit with different poly A tail lengths. The encoded protein itself is a 135-amino acid protein (15 kDa), with no apparent homology to any other known protein. Next the protein was recloned into E. coli using the pET-15b-vector. Upon isopropyl-1-thio-beta-D galactopyranoside induction, the recombinant protein, rNecH1, could be recovered by urea treatment from inclusion bodies. The rNecH1 protein binds to different collagens: human I > rat I > human III = calf skin I in a specific, dose dependent, and saturable manner. PMID- 11426728 TI - Plasmodium malariae blood-stage dynamics. AB - We examine the dynamics of parasitemia, fever, and gametocytemia reflected in the preintervention charts of 180 malaria-naive U.S. neurosyphilis patients infected with the USPHS strain of Plasmodium malariae, for malariatherapy, focusing on the 84 charts for which more than 35 days of patency preceded intervention and daily records encompassed 92% or more of the duration of each infection. Inoculum size did not influence any outcome variable. Fevers (days with temperatures > or =101 F) followed patterns that fit recognized brood structures more often than did our approximations of merogony cycles (via local peaks in parasitemia), but neither closely fit textbook quartan patterns. There were no discernable patterns in gametocytemia. Successful transmission to mosquitoes increased following subcurative drug treatment but did not depend on detectable gametocytemia. PMID- 11426729 TI - Thaumasioscolex didelphidis n. gen., n. sp. (Eucestoda: Proteocephalidae) from the black-eared opossum Didelphis marsupialis from Mexico, the first proteocephalidean tapeworm from a mammal. AB - Thaumasioscolex didelphidis n. gen., n. sp. is described from the intestine of the black-eared opossum Didelphis marsupialis L. (Marsupialia: Didelphidae) from Los Tuxtlas, Veracruz, Mexico. The new genus differs from all proteocephalidean genera in the morphology of the scolex that is formed by 4 well separated lobes each containing 1 noncircular sucker opening laterally inside the exterolateral cavity, a large-sized body (length up to 1 m), a large number of testes, the shape of gravid proglottids that are inversely craspedote (the anterior border of a proglottid overlaps the posterior border of a preceding proglottid), eggs in groups mostly of 4-6 eggs each, and an embryophore bearing digitiform projections on its external surface. This is the first tapeworm of the Proteocephalidea, the members of which were previously reported exclusively from poikilotherm vertebrates (freshwater fishes, amphibians, and reptiles), found in a homoiotherm vertebrate. PMID- 11426730 TI - Spinitectus osorioi n. sp. (Nematoda: Cystidicolidae) from Chirostoma spp. (Osteichthyes: Atherinidae) in Lake Patzcuaro, Michoacan, Mexico. AB - Spinitectus osorioi (Nematoda: Cystidicolidae) is described from the freshwater atherinids Chirostoma estor and Chirostoma attenuatum from Lake Patzcuaro in the Mesa Central, Michoacan State, Mexico. This nematode is characterized by a conspicuous protuberance on the ventral surface of the distal end of the long spicule that distinguishes it from its congeners in North America and in the neotropics. In addition, the species can be readily distinguished from 4 of the 5 nominal species of North American freshwater Spinitectus by the absence of either a terminal barb or heel on the short spicule and from Spinitectus mexicanus by the spination. Previous records of Spinitectus carolini from Chirostoma spp. in Mexico (Lakes Patzcuaro and Zirahuen) refer to S. osorioi, and the species appears to be specific to Chirostoma spp. The geological history of the Mesa Central drainages and the historical biogeography of freshwater atherinids in this region suggest that the origin of S. osorioi may be associated with either the marine history of their hosts or with host-switching from more distantly related freshwater hosts after colonization of freshwater environments by atherinids. PMID- 11426731 TI - Vexillata liomyos n. sp. (Nemata: Ornithostrongylidae) from Liomys pictus (Rodentia: Heteromyidae) from Mexico, with comments on the synlophe of Vexillata armandae. AB - Individuals of a new species of Vexillata were collected from the small intestines of Liomys pictus from the Estaci6n de Biologia Chamela, in Jalisco State, Mexico. The new species shows an array of characters that allow us to recognize it as a member of Vexillata; however, it can be distinguished from other species of the genus in that males possess an asymmetrical caudal bursa, females possess a characteristic cuticular inflation at the level of the ovijector, and both sexes possess a synlophe with 9 ridges at the midbody. Additional detail of the synlophe of Vexillata armandae Gardner et al., 1994 from Chaetodipus hispidus in New Mexico shows that both sexes have 12 cuticular ridges just posterior to the cephalic inflation, and in the posterior region of the body, females have 9 ridges of equal size while males possess 11 equal-sized ridges. In both sexes, the carene disappears at the posterior end of the body. PMID- 11426732 TI - Toxocara malaysiensis n. sp. (Nematoda: Ascaridoidea) from the domestic cat (Felis catus Linnaeus, 1758). AB - Toxocara malaysiensis n. sp. from the small intestine of the domestic cat (Felis catus L.) in Malaysia is described and illustrated. This ascaridoid nematode was previously assumed to be Toxocara canis, which it superficially resembles, or designated Toxocara sp. cf. canis. The new species differs from T. canis in the shape of the cervical alae in cross section, spicule length, and the lip structure. It is also distinct from other species assigned to Toxocara. PMID- 11426733 TI - Anindobothrium n. gen. (Eucestoda: Tetraphyllidea) inhabiting marine and freshwater potamotrygonid stingrays. AB - Anindobothrium n. gen. is proposed to accommodate Caulobothrium anacolum inhabiting Himantura schmardae from Colombia, and 2 new species, one inhabiting Potamotrygon orbigny in Brazil and the other inhabiting Paratrygon aereiba in Venezuela. Members of the new genus resemble members of Pararhinebothroides, Rhinebothroides, and Anthocephalum by having bothridia with poorly differentiated apical suckers and vasa deferentia expanded into external seminal vesicles. It further resembles Pararhinebothroides, Rhinebothroides, and Anthocephalum cairae by having vas deferens inserted near the poral rather than aporal end of the cirrus sac. The 3 species assigned to the new genus form an apparent monophyletic group, based on the possession of 3 putative synapomorphies: (1) genital pores in the anterior 1/4 of the proglottid, a trait that is unusual, but not unique, among phyllobothriids; (2) anteroventral ovarian lobes converging to the center of the proglottid, a character not previously reported for phyllobothriids; and (3) ovarian lobes comprising a loose network of digitiform processes. PMID- 11426734 TI - Two new species of Macrobothridium (Cestoda: Diphyllidea) from rhinobatid elasmobranchs in the Gulf of Gabes, Tunisia, with notes on the status of the genus. AB - As a result of an investigation of metazoan parasites of elasmobranch fishes in the Gulf of Gabes, Tunisia, we discovered 2 new species of diphyllidean cestodes. Macrobothridium euterpes n. sp. is described from the spiral intestine of Rhinobatos rhinobatos, and Macrobothridium syrtensis n. sp. from the spiral intestine of Rhinobatos cemiculus. Macrobothridium euterpes is distinguished from the only other species in the genus (Macrobothridium rhynchobati) by the number of rostellar hooks, size, genital pore position, vagina position, and ovary shape. Macrobothridium syrtensis is distinguished from M. rhynchobati by the hook morphology, testis number, and overall size, and from M. euterpes by the number of rostellar hooks, testis number, genital pore position, vagina position, and ovary shape. These are the first 2 species added to the genus since its establishment in 1989. A standardized formula for representing the number and arrangement of rostellar hooks in diphyllidean species is presented. PMID- 11426735 TI - Babesia leo n. sp. from lions in the Kruger National Park, South Africa, and its relation to other small piroplasms. AB - Babesia leo, a small piroplasm isolated from lions in South Africa is described as a distinct species based on a phylogenetic analysis of the 18S rRNA gene. Intraerythrocytic trophozoite and merozoite stages of B. leo are morphologically indistinguishable from other small piroplasms of felids. Previous studies showed that B. leo was biologically and antigenically distinct from B. felis, which is known to infect wild and domestic felids in South Africa. Molecular characterization showed strong support for the phylogenetic seperation of B. leo as a distinct species from B. felis and other felid piroplasms. Phylogenetic analysis also showed that Babesia microti and all of the felid piroplasms from Africa with known 18S rRNA gene sequences available, including B. leo, formed a single, separate clade, sister to the other babesial and theilerial piroplasm parasites. PMID- 11426736 TI - Glypthelmins poncedeleoni n. sp. (Trematoda: Macroderoididae) of amphibians from the neotropical region of Mexico. AB - A new species of Glypthelmins (Trematoda: Macroderoididae) is described from the intestine of Bufo marinus and Leptodactylus melanonotus from several localities of the Neotropical Region of Mexico. Glypthelmins poncedeleoni n. sp. can be differentiated from other species of the genus by having extracecally distributed vitellaria, ovary always larger than testes, and vitelline follicles grouped in 7 post-testicular, nonoverlapping, rosette-like clusters and 5 pretesticular overlapping clusters. PMID- 11426737 TI - Comparison of ivermectin, doramectin, selamectin, and eleven intermediates in a nematode larval development assay. AB - Chemical substitutions at pharmacologically relevant sites such as C-5, C-13, C 22,23, and C-25 were examined in ivermectin, doramectin, selamectin, and a series of 11 other intermediates using a larval development assay with Haemonchus contortus. A range of activities spanning 5 orders of magnitude were manifest with small changes in the substituents to the 14 avermectins. Within this compound series, there was no major potency advantage or disadvantage to a disaccharide over a monosaccharide substituent at C-13. Ivermectin and doramectin were each fully effective at a concentration of 0.001 microg/ml, and both were similar to their respective monosaccharide homologs. Specific patterns emerged among the analogs with substituents at C-5. Analogs possessing hydroxyl groups at C-5 were superior in activity by several orders of magnitude over those with oxo substituents. Replacement of the oxo with an oxime (NOH) restored activity to some degree but did not restore it to the level of those possessing the hydroxyl substituent. Consequently, ivermectin and doramectin that possess hydroxyl moieties at C-5 were superior against H. contortus to those like selamectin that have oxime substituents. There was no advantage for analogs with a single or double bond at C-22,23 within the cyclohexyl series, and these analogs had equivalent activity as those with a single bond at C-22,23 in the sec butyl/isopropyl series. However, there was superior activity for the analog series that possessed the combination of a double-bond at C-22,23 and a sec butyl/isopropyl substituent at C-25. As a result, the most potent compound in this test was not any of the 3 commercialized avermectins but was a monosaccharide with a double bond at C-22,23, an hydroxyl at C-5, and a sec butyl/isopropyl moiety at C-25. PMID- 11426738 TI - Immunoprophylaxis against Fasciola hepatica in rabbits using a recombinant Fh15 fatty acid-binding protein. AB - Previous studies of ours have demonstrated that a recombinant protein (Fh15) related to fatty acid-binding proteins did not induce significant protection in rabbits challenged 2 or 4 wk postimmunization over nonimmunized controls. In the current study, rabbits were immunized with Fh15 and challenged with Fasciola hepatica metacercariae 12 and 20 wk later. In the current study in which longer lag periods for challenge infection after the second immunization were used, worm burden reductions compared to adjuvant controls were a significant 43% and 76%, respectively. Importantly, rabbits immunized with Fh15 had significant numbers of immature flukes, 66% in the 12-wk period and 84% in the 20-wk lag period as compared to controls. In addition, liver lesions were clearly diminished in the vaccinated rabbits. Enzyme-linked immunosorbent assay absorbance values showed that immunized rabbits developed high antibody levels to Fh15 from 8 wk after the first immunization and did not increase after challenge. These results suggest that a recombinant F. hepatica molecule related to fatty acid-binding proteins induces protective (worm burden reductions), anti-fecundity (immature flukes), and anti-pathology (less liver lesions) effects in rabbits and may serve as a model for the immunoprophylaxis of fascioliasis. PMID- 11426739 TI - Modifications of Kohn's chlorazol black E staining and Wheatley's trichrome staining for temporary wet mount and permanent preparation of Entamoeba histolytica. AB - Preparation of stained smears of Entamoeba histolytica has several drawbacks. We therefore tried to simplify the staining procedures by modifing Kohn's chlorazol black E staining and Wheatley's trichrome staining techniques. Trophozoites and cysts of axenically cultured E. histolytica and Entamoeba invadens, respectively, and trophozoites and cysts of E. histolytica in stools of patients were used. Karyosomes and peripheral chromatin of nuclei and chromatoid bodies became distinctly visible after amoebae were suspended in the basic solution of Kohn's stain. Amoebae fixed in suspension with either basic solution or Bouin's fixative were clearly stained with Kohn's and trichrome preparations, both as wet mounts directly and as permanent slides after processing for mounting. These procedures were easier when the basic solution was used as a fixative and trichrome stain was employed. Erythrocytes ingested by trophozoites, however, were not stained with either of these preparations after fixation in the basic solution but were clearly stained when Bouin's fixative was used. Cysts of E. histolytica in stools concentrated using basic solution (instead of formalin) and ether were also stained with these stains. Consequently, without employing highly toxic mercuric chloride, wet mounts and permanent smears can be prepared with permanent stains, and preserved cysts can be stained after concentration. PMID- 11426740 TI - Human T and B cell immunoreactivity to a recombinant 23-kDa Cryptosporidium parvum antigen. AB - Cryptosporidial infection in humans results in parasite-specific IgG, IgM, and IgA antibody responses, but little is known of the cell-mediated immune responses to cryptosporidial antigens. In a convenience sample of 35 Haitian residents, there was a high level of cryptosporidial exposure (>90%) as determined by immunoblot reactivity of serum against cryptosporidial antigens. An attempt was made to determine if there was a relationship between antibody and T cell mediated responses to recombinant Cp23 antigen and how this correlated with reactivity to crude sporozoite antigen preparations (SAg). T cell reactivity was greater against SAg (57%) than to Cp23 (34.3%) as measured by [3H]thymidine incorporation. Proliferative responses to Cp23 were significantly correlated with SAg responses. By enzyme-linked immunosorbent assay, most persons had IgG responses to both SAg (91.4%) and to recombinant Cp23 (88.5%). Antibody responses were greater among persons who exhibited T cell responses to SAg and Cp23. This study demonstrates that recombinant Cp23 antigen could be a useful antigen for detection of both antibody and cell-mediated responses in epidemiologic studies. PMID- 11426741 TI - The pig as a host for Schistosoma mekongi in Laos. AB - A survey of helminths in domestic pigs was conducted in Khong District, Laos, to elucidate if these domestic animals could act as definitive hosts for Schistosoma mekongi and to obtain a general overview of their helminthological infection status. Fecal samples were collected from 98 pigs. Twelve pigs (12.2%) were found to excrete S. mekongi eggs. Infection was confirmed by detection of S. mekongi eggs in tissues of liver, rectum, and cecum of 2 pigs. A total of 75.8% of the pigs was infected with 1 or more helminth species. This study showed that pigs may act as a definitive host for S. mekongi. PMID- 11426742 TI - Unusual electrophoretic patterns for phosphoglucomutase and fumarase in a population of Lecithochirium rufoviride (Trematoda: Hemiuridae), a parasite of Conger conger. AB - Electrophoretic analyses of phosphoglucomutase (PGM) and fumarase (FH) in a population of Lecithochirium rufoviride parasitizing Conger conger, revealed 2 independent activity zones for each enzyme on starch gel electrophoresis. However, some individuals exhibited only 1 activity zone for 1 or both enzymes. The banding patterns observed strongly suggest that (1) PGM is coded by 2 polymorphic loci, Pgm-1 (expressed in all individuals) with allelic frequencies not significantly different from those expected under Hardy-Weinberg equilibrium, and Pgm-2 (expressed in a subset of individuals); and (2) FH is coded by 2 loci, Fh-2 (monomorphic and expressed in all individuals) and Fh-1 (expressed in a subset of individuals). A high degree of concordance (88.75%) was observed between the expression and nonexpression of Pgm-2 and Fh-1. The most likely explanations for these findings are either variation in enzyme expression with developmental stage or the presence of null alleles at high frequencies in the population. PMID- 11426743 TI - The anatomic basis for lung cancer staging: the end of the beginning? PMID- 11426744 TI - Prognosis and survival after resection for bronchogenic carcinoma based on the 1997 TNM-staging classification: the Japanese experience. AB - BACKGROUND: A new TNM staging system was proposed, and the previous system was revised in 1997. METHODS: To evaluate the new TNM staging system for lung cancer, records of 3,043 lung cancer patients who underwent pulmonary resection at the National Cancer Center Hospital, Tokyo, were analyzed. RESULTS: With regard to clinical stages, 3 patients had occult carcinoma; 786 patients had stage IA disease; 759 patients, stage IB; 54 patients, stage IIA; 469 patients, stage IIB; 582 patients, stage IIIA; 211 patients, stage IIIB; and 179 patients, stage IV. The 5-year survival rates for the respective stages were 70.8% for stage IA, 44.0% for stage IB, 41.1% for stage IIA, 36.9% for stage IIB, 22.7% for stage IIIA, 20.1% for stage IIIB, and 21.6% for stage IV. In terms of postoperative stages, 7 patients were classified in stage 0, 610 in stage IA, 506 in stage IB, 114 in stage IIA, 432 in stage IIB, 702 in stage IIIA, 448 in stage IIIB, and 224 in stage IV. The 5-year survival rates were as follows: stage IA, 79.0%; stage IB, 59.7%; stage IIA, 56.9%; stage IIB, 45.0%; stage IIIA, 23.6%; stage IIIB, 16.5%; and stage IV, 5.1%. CONCLUSIONS: In the clinical stage, there were significant prognostic differences between stage IA and stage IB, stage IIB and IIIA, and stage IIIA and stage IIIB, but there was no significant difference in 5 year survival rates between stage IB and stage IIA, stage IIA, and IIB, and stage IIIB and stage IV. In the postoperative stage, there were significant differences in 5-year survival rates between each stage except for stage IB and stage IIA. PMID- 11426745 TI - Prognostic implication of Ki-67 immunostaining in treating subclinical pleural cancer found at thoracotomy in lung cancer patients. AB - BACKGROUND: Therapeutic principles for managing subclinical pleural cancer found unexpectedly during intraoperative examination are unclear. We analyzed prognostic factors including the tumor proliferative marker Ki-67 in these circumstances. METHODS: The cases of 65 surgically treated patients with lung cancer and subclinical T4 pleural cancer, microscopic in 25 and macroscopic in 40, were reviewed. RESULTS: The overall 5-year survival rate of patients undergoing lobectomy was 14.3%. For patients with T4 NO disease, the 5-year survival rate was 46.7%. In patients with a low Ki-67 labeling index, the 5-year survival rate was 28.6%. The Ki-67 labeling index was a significant (p < 0.05) indicator of survival. Multivariate analysis demonstrated Ki-67 labeling index, lymph node involvement, and tumor differentiation to be the most influential prognostic factors for postoperative survival (p < 0.01). CONCLUSIONS: In the treatment of lung cancer patients with subclinical pleural cancer found at thoracotomy, tumor resection is not necessarily contraindicated. Resection appears to be beneficial in patients with no nodal involvement or a low tumor Ki 67 labeling index. This index is a good therapeutic indicator for lung cancer patients. PMID- 11426746 TI - Nodal occult metastasis in patients with peripheral lung adenocarcinoma of 2.0 cm or less in diameter. AB - BACKGROUND: Detection of occult micrometastasis in regional lymph nodes is crucial for diagnosis and selection of appropriate therapy for patients with pN0 non-small-cell lung carcinoma. Using immunohistochemical staining, we evaluated the impact of detection of occult micrometastasis on the prevalence and prognosis of patients with lung adenocarcinoma of 2.0 cm or less in diameter. METHODS: A total of 103 pN0 disease patients with peripheral lung adenocarcinomas of 2.0 cm or less in diameter were enrolled in this study. We studied 1,438 regional lymph nodes for occult micrometastasis by immunohistochemical staining for cytokeratins. RESULTS: Micrometastasis was detected in 49 lymph nodes (3.4%) of 21 patients (20.4%) but not in patients with localized bronchioloalveolar carcinoma or localized bronchioloalveolar carcinoma with foci of collapse of alveolar structure. The 5-year survival rate (61.9%) of patients with micrometastasis was significantly (p = 0.0041) lower than that of patients without micrometastasis (86.3%). CONCLUSIONS: There still remains a risk of nodal micrometastasis in patients with primary peripheral lung adenocarcinoma, even if the diameter of the tumor is smaller than 2.0 cm. Selection of patients for limited surgery should be done prudently, taking into consideration the risk of nodal micrometastasis. PMID- 11426747 TI - Open-lung biopsy guides therapy in children. AB - BACKGROUND: Open-lung biopsy is uncommon in children. Modern indications and outcomes are unknown. METHODS: This is a retrospective review of 64 open-lung biopsies (58 patients) from 1976 to 1996. Open-lung biopsies were used to grade vasculopathy in 8 patients (12% of 64) with pulmonary hypertension and in 10 patients (16% of 64) with combined pulmonary hypertension and lung parenchymal disease. Forty-six biopsies (72%) were obtained to diagnose parenchymal disease. Comparisons were made between biopsies performed from 1976 to 1989 and from 1990 to 1996. RESULTS: In the period 1990 to 1996, there were significantly more infants (p = 0.03), comorbid disease (p = 0.009), extracorporeal membrane oxygenation support (p < 10(-4)), and ventilator dependence (p = 0.05) and significantly less immunocompromise (p = 0.04). A definitive diagnosis was made in 43 of 64 cases (67%) and altered workup in 63 of 64 cases (98%). No correlation existed between Heath-Edwards grade of microangiopathy and catheterization data. Definitive diagnosis was most strongly associated with a nonimmunocompromised patient (p < 10(-4)). Although only one death (1.5%) was related to open-lung biopsy, the procedure was associated with a 30% inhospital mortality rate and an 11% morbidity rate. Of the 19 deaths, 1 patient died from the procedure, 13 died from their diseases, and 5 had support withdrawn. Death was associated with preoperative ventilator dependence (p < 10(-4)) and extracorporeal membrane oxygenation (p = 0.007). CONCLUSIONS: Pediatric open-lung biopsy commonly alters the diagnostic workup (98%). It is recommended for children who have been supported for 2 weeks by extracorporeal membrane oxygenation and for those with combined pulmonary hypertension and parenchymal lung disease. It is less useful in immunocompromised children. PMID- 11426748 TI - Bile exposure of the denervated stomach as an esophageal substitute. AB - BACKGROUND: Both the supine position and the existence of a gastric drainage procedure are suspected to promote reflux of duodenal juice into the denervated intrathoracic stomach. Erythromycin has been shown to weaken pyloric resistance to gastric outflow and to enhance antral motility, gastric emptying, and gallbladder contractility. METHODS: The presence of bile in the gastric transplant of 79 patients was monitored over a 24-hour period with use of the Bilitec 2000 optoelectronic device 3 to 195 months after subtotal esophagectomy. Ten patients were reinvestigated after a 3-year period. Five groups were studied: group I: n = 12, no gastric drainage, never given erythromycin, group 2: n = 40, gastric drainage, never given erythromycin, group 3: n = 7, no gastric drainage, given erythromycin, group 4: n = 13, gastric drainage, given erythromycin, and group 5: n = 7, no longer given erythromycin (with or without gastric drainage). The percentage of time gastric bile absorbance was more than 0.25 was calculated for the total, supine, and upright periods of recording in reference to data from 25 healthy volunteers. RESULTS: The Bilitec test was pathologic in 9 of the 12 patients of group 1 whereas it was normal in three. Gastric exposure to bile was longer in group I patients than in controls for the total (p = 0.012) and supine (0.036) periods, but the difference did not reach statistical significance for the upright period (p = 0.080). Bile exposure in group 4 did not significantly differ from controls (total: p = 0.701; supine: p = 0.124; upright: p = 0.712). Bile exposure for the total period did not significantly differ whether patients were taking erythromycin or the drug had been discontinued at the time of the study (p = 0.234); and it tended to decrease with time in patients investigated twice (p = 0.046). CONCLUSIONS: Gastric exposure to bile after truncal vagotomy and transposition of the stomach up to the neck is pathologic in three quarters of patients. It is more marked in the supine than in the upright position and tends to decrease with time. The addition of a gastric drainage procedure in combination with erythromycin therapy tends to normalize gastric exposure to bile. The effects of erythromycin may persist after discontinuation of the drug. PMID- 11426749 TI - Oropharyngeal dysphagia after cardiac operations. AB - BACKGROUND: As many as 15% of hospitalized patients have oropharyngeal dysphagia. The incidence and causes of postoperative oropharyngeal dysphagia (OD) in patients having cardiac operations are poorly documented and the best treatment is uncertain. We undertook a study to evaluate OD in patients having cardiac operations. METHODS: As part of a quality improvement project, all patients operated on in 1998 and 1999 were monitored for the signs or symptoms of OD. Patients with OD had diagnostic and therapeutic interventions to limit adverse outcomes. At the end of the 2-year evaluation period, patient risk factors, diagnoses, results of interventions, and outcomes were measured. RESULTS: Thirty one out of 1,042, patients (3%) had OD. OD is more common in older patients (p < 0.0001) with diabetes (p = 0.02), renal insufficiency (p = 0.012), hyperlipidemia (p = 0.046), and preoperative congestive heart failure (p < 0.0001), and in those having noncoronary artery bypass procedures (p < 0.0001). One patient with OD died from respiratory arrest, presumably secondary to aspiration. Modified barium swallow (MBS) identified oral dysphagia in 2 patients, pharyngeal dysphagia in 7 patients, and both oral and pharyngeal dysphagia in 17 patients. One patient had a structural defect (cervical osteophyte) causing dysphagia and 4 patients had no identifiable cause of dysphagia on MBS. Postoperative neurologic complications are more common in patients with OD. Ten of 31 patients (32%) with OD had some new neurologic complication after operation compared with 36 of 1,011 (3.5%) who had a postoperative neurologic problem without OD. In 19 patients with OD no cause for swallowing difficulty was identified. Specifically, no metabolic, myopathic, or infectious abnormalities were identified in any patient with OD. Hospital charges were significantly increased in patients with OD ($69,320 versus $36,087, p < 0.0001). Therapy consisting of modification of eating behavior and swallowing technique and in some severe cases enteral or parenteral feeding was successful in all patients except 1, but 4 patients required more than 4 months of supportive care before return to oral feeding was possible. CONCLUSIONS: OD is associated with increased cost and morbidity. Older patients with diabetes, preoperative heart failure, and renal insufficiency are at increased risk for OD. Early recognition and intervention is likely to result in satisfactory outcome but may be associated with a protracted postoperative course. PMID- 11426750 TI - Results of expandable metal stents for malignant esophageal obstruction in 100 patients: short-term and long-term follow-up. AB - BACKGROUND: Expandable metal stents palliate malignant dysphagia in most cases, but early complications and outcomes in long-term survivors have not been well described. This report summarizes our experience with expandable metal stents for malignant dysphagia. METHODS: Over a 48-month period, 127 stents were placed in 100 patients with dysphagia from esophageal cancer (93%) or lung cancer. Most had undergone prior treatment. Dysphagia scores, duration of palliation, complications, and reintervention were evaluated. RESULTS: Immediate improvement in dysphagia was observed in 85% of patients with no procedure-related deaths. Dysphagia score decreased from 3.3 before stent to 2.3 (p < 0.005). Average interval to reintervention was 80 days. In 40 patients surviving more than 120 days, 31 (78%) required reintervention. Major complications occurred in 3 patients receiving poststent chemoradiation (tracheoesophageal fistula, T1 vertebral body abscess, mediastinal abscess). Other complications included unsatisfactory deployment requiring immediate removal (3 patients), migration (11 patients), pain requiring removal (2 patients), food impaction (10 patients), and tumor ingrowth (37 patients). CONCLUSIONS: Expandable metal stents offer excellent short-term palliation of malignant dysphagia. In long-term survivors, recurrent dysphagia requiring reintervention is common. In a small subset of patients receiving chemoradiation after stent placement, major complications were observed. PMID- 11426751 TI - Ivor Lewis esophagogastrectomy for esophageal cancer. AB - BACKGROUND: To examine the efficacy of the Ivor Lewis esophagogastrectomy for esophageal carcinoma prior to the widespread use of preoperative chemotherapy and irradiation, we reviewed our experience. METHODS: We reexamined the cases of 220 consecutive patients who underwent an Ivor Lewis esophagogastrectomy for esophageal cancer from January 1992 through December 1995. RESULTS: There were 196 men (89.1%) and 24 women. Median age was 65 years (range, 29 to 85 years). The results of pathological study showed adenocarcinoma in 188 patients (85.5%), squamous cell carcinoma in 31 (14.1%), and leiomyosarcoma in 1 patient (0.5%). Postsurgical staging was as follows: stage 0 in 10 patients, stage I in 19, stage IIa in 38, stage IIb in 28, stage III in 111, and stage IV in 14. The operative mortality rate was 1.4% (3 patients), and complications occurred in 83 patients (37.7%). Follow-up was 98.6% complete. Median survival for operative survivors was 1.9 years (range, 32 days to 8.7 years). The overall 5-year survival rate was 25.2%; it was 80% for patients in stage 0, 94.4% for those in stage I, 36.0% for those in stage IIa, 14.3% for patients in stage IIb, 10% for those in stage III and 0% for patients in stage IV. CONCLUSIONS: Ivor Lewis esophagogastrectomy for esophageal cancer is a safe operation. Long-term survival is stage dependent. The low survival associated with advanced cancers should stimulate the search for effective neoadjuvant therapy. PMID- 11426752 TI - Effective treatment of malignant pleural effusion by minimal invasive thoracic surgery: thoracoscopic talc pleurodesis and pleuroperitoneal shunts in 101 patients. AB - BACKGROUND: For effective palliation of patients with malignant pleural effusion due to advanced neoplastic disease, any proposed treatment should have low procedure-related mortality and morbidity. METHODS: The clinical outcome of 119 thoracoscopies in 101 patients (56 women, 45 men), from 42 to 91 years of age (mean, 68 +/- 9 years) with malignant pleural effusions was evaluated in a retrospective study. Video-assisted thoracoscopy (VATS) talc pleurodesis was done in 105 instances, and a pleuroperitoneal shunt was performed 14 times as an alternative when complete expansion of the lung could not be achieved due to tumor implants on the visceral pleura. RESULTS: The VATS talc pleurodesis resulted in clinically significant improvement of dyspnea in 92.2% of the patients. Thirty-day mortality was 2.8% and morbidity was 2.8%. The mean duration of postoperative survival was 6.7 months. Recurrent pleural effusion occurred in 5.7% of patients after a mean interval of 6 months. Clinical relief of dyspnea was obtained in 73% of the patients treated with pleuroperitoneal shunts. Thirty day mortality in this group was 21% and morbidity was 14.3%. The mean duration of survival was 4.2 months. CONCLUSIONS: The VATS talc pleurodesis is appropriate for palliation of patients with malignant pleural effusions and should be performed once the diagnosis has been confirmed. Patients with lungs trapped by visceral carcinomatosis may benefit from placement of a pleuroperitoneal shunt as an alternative. PMID- 11426753 TI - Thoracoscopic decortication: a role for video-assisted surgery in chronic postpneumonic pleural empyema. AB - BACKGROUND: We evaluated a technique of video-assisted thoracoscopic (VAT) decortication of the visceral cortex to reexpand entrapped lung in cases of chronic postpneumonic pleural empyema. METHODS: A prospective cohort study of 48 consecutive patients with multiloculated postpneumonic pleural empyema in whom visceral pleural decortication was required was studied. The effect of VAT decortication on perioperative outcome and factors affecting its success were assessed. RESULTS: Before the introduction of VAT decortication 12 patients were treated by thoracotomy (group T). In the subsequent 36 patients VAT decortication was attempted with success in 21 (group VS) but lung expansion was not observed in 15 patients (group VF) who required thoracotomy. There was no difference in the age or sex distribution of the 3 groups. Operating time was significantly longer in group T than group VS, mean difference 30.3 minutes (p = 0.001) and postoperative hospital stay was longer in group T than group VS, mean difference 2.9 days (p = 0.004). The success of VAT decortication was not related to either the delay between onset of symptoms or hospital admission and surgery; indeed the operating time decreased with increasing preoperative delay. However, success was related to increasing operative experience (p = 0.001). CONCLUSIONS: VAT decortication is a feasible new technique to achieve lung reexpansion in chronic postpneumonic pleural empyema and has perioperative benefits over thoracotomy. PMID- 11426754 TI - Efficient naked plasmid cotransfection of lung grafts by extended lung/plasmid exposure time. AB - BACKGROUND: Multiple gene cotransfection may be an effective strategy to modulate concurrent pathologic events after lung transplantation. We investigated in vivo naked plasmid lung cotransfection during cold preservation and the role of lung parenchyma/naked plasmid exposure time. METHODS: F344 rats underwent left main bronchus instillation of pCF1-CAT (chloramphenicol acetyl transferase) (130 microg) +/- pCF1-beta-Gal (beta-galactosidase) (130 microg) in saline. Part Ia: 4 degrees C preservation versus cotransfection. Lung isografts (4 groups, n = 8) were stored after transfection for 1 (2 groups: one received only pCF1-CAT), 6, and 18 hours. Recipient sacrifice was after 48 hours. Part Ib: 4 degrees C preservation versus transgene expression. Rats were sacrificed 48 hours after transfection in a nontransplant setting (2 groups, n = 8; one received only pCF1 CAT). In a third group (n = 8) lungs were harvested 24 hours after transfection, stored for 18 hours, and recipients were sacrificed after 24 hours. The CAT and beta-Gal enzymatic-linked immunosorbent assays were performed. Part II: Lung/plasmid exposure time. In three groups (n = 6) after pCF1-CAT transfection the left main bronchus was not clamped, clamped for 10 minutes, or clamped for 1 hour. Sacrifice was after 48 hours. RESULTS: Part Ia: Lung CAT protein was (in picograms per 100 microg of total protein): median, 42 (range, 25 to 95) after 1 hour (only CAT); 67 (19 to 296) after 1 hour, 32 (6 to 157) after 6 hours; and 9 (5 to 243) after 18 hours. Lung beta-Gal protein was (in picograms per 100 microg of total protein): median, 20 (range, 5 to 353) after 1 hour; 17 (6 to 157) after 6 hours; 4 (1 to 74) after 18 hours (1 hour versus 18 hours, p = 0.04 for both proteins). CAT and beta-Gal production were significantly correlated (p = 0.0001, r = 0.924). Part Ib: Lung CAT protein was (in picograms per 100 microg of total protein): median, 2 (range, 0.6 to 10) no transplant, only CAT; 7 (0.3 to 13) no transplant; 3 (0.9 to 14) transplant. Part II: Left lung CAT protein was (in picograms per 100 microg of total protein): median, 31 (range, 6 to 83) no clamp; 74 (25 to 430) 10 minutes of clamp; 111 (30 to 263) 1 hour of clamp. Right lung CAT protein was (in picograms per 100 microg of total protein): median, 0.06 (range, 0 to 0.9) no clamp; 1 (0 to 6) 10 minutes of clamp; 1 (0 to 18) 1 hour of clamp. CONCLUSIONS: Efficient lung isograft endobronchial cotransfection results from using naked plasmid. Cold preservation affects transfection efficiency but not transgene expression. Lung parenchyma/naked plasmid exposure time determines transfection efficiency. PMID- 11426755 TI - Association of IL-8 and MCP-1 with the development of reexpansion pulmonary edema in rabbits. AB - BACKGROUND: The aim of this study is to determine the relationships between the cytokines and the inflammatory response in reexpansion pulmonary edema (RPE). METHODS: We examined the cell population, epithelial permeability measured by Evans blue dye (EB), betaglucuronidase and cytokine concentrations in bronchoalveolar lavage fluid (BALF) and/or blood using a rabbit RPE model. RESULTS: We confirmed that RPE is characterized by recruitment of polymorphonuclear leukocytes (PMNs), the release of PMN granular contents into the air spaces, and increased vascular permeability. These findings were highly correlated with increased interleukin-8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1) concentrations in the BALF. Growth related oncogene (GRO) was detected in the BALF from only 2 of the 7 reexpanded lungs while TNFalpha was not detected in any rabbits. A similar but less severe inflammatory response to the reexpanded lung was found in the contralateral lung. CONCLUSIONS: IL-8 and MCP-1 may play important roles in the development of RPE; the inflammatory response is independent of TNFalpha and unilateral reexpansion of the lung induces an inflammatory response not only in the reexpanded lung but also in the contralateral lung. PMID- 11426756 TI - The paradox of survival results after heart transplantation for cardiomyopathy caused by Trypanosoma cruzi. First Guidelines Group for Heart Transplantation of the Brazilian Society of Cardiology. AB - BACKGROUND: Donor supply limits heart transplantation (HT) and relative priority should be given to cases with greater chances of success. The objectives of this multicenter study were (1) to determine the survival rate after heart transplantation for patients with Chagas' heart disease (ChHD) in comparison with other causes; and (2) to identify the causes of death specifically due to reactivation of the Trypanosoma cruzi infection. METHODS: We studied 720 patients who had undergone orthotopic heart transplantation and were followed in 16 heart transplantation centers. The etiology was idiopathic dilated cardiomyopathy in 407 patients, ischemic cardiomyopathy in 196 patients, and ChHD in 117 patients. RESULTS: Follow-up was 2.87 +/- 3.05 years (from 1 month to 13.85 years). Survival of ischemic recipients at 1, 4, 8, and 12 years was 59%, 44%, 34%, and 22%, respectively; for idiopathic dilated cardiomyopathy it was 69%, 57%, 40%, and 32%; and for ChHD it was 71%, 57%, 55%, and 46% (p < 0.027). In ischemic recipients the most frequent causes of death were infection (15.3%), acute graft failure (13.3%), and graft coronary artery disease/sudden death (7.7%). In idiopathic dilated cardiomyopathy the causes were infection (11.1%), rejection (9.6%), and acute graft failure (9.1%). In ChHD the causes were infection (10.3%), rejection (10.3%), and neoplasm (4.3%). In ChHD, reactivation of the cruzi infection was the cause of death in 2 patients. CONCLUSIONS: The survival results after heart transplantation are paradoxical according to the usually high expected death rates for Chagas' disease. Heart transplantation for ChHD should be regarded as a valuable treatment option. PMID- 11426757 TI - Risk analysis in patients bridged to transplantation. AB - BACKGROUND: Efforts to predict mortality in bridge to cardiac transplant patients have concentrated on preventricular assist device (VAD) status. To more fully identify factors influencing survival to transplant, we reviewed the preoperative and postoperative VAD courses of 105 bridge to transplant patients. METHODS: Sixty-four parameters (34 pre-VAD, 30 post-VAD), including hemodynamics, complications, and evaluations of major organ function were examined and analyzed. RESULTS: Thirty-three patients (31%) died on VADs and 72 were transplanted. There were two posttransplant operative deaths (3%). By univariate analysis 23 of 64 factors were significant. These 23 factors were entered into a stepwise logistic regression analysis to identify predictors of survival to transplant. Four factors, including pre-VAD intubation (p < 0.005), cardiopulmonary bypass (CPB) time during VAD insertion (p < 0.0001), mean pulmonary artery pressure (first postoperative day after VAD) (p < 0.0002), and highest post-VAD creatinine (p < 0.01) were independent predictors of transplantation. CONCLUSIONS: Other than the need for intubation, pre-VAD variables were of little value in predicting survival to transplant. Problems during VAD insertion (long CPB time) and post-VAD renal insufficiency were independent predictors. Severe complications that developed during the interval of VAD support, including cerebrovascular accident, bleeding and infection, were surprisingly not predictors for transplantation. PMID- 11426758 TI - Reappearance of myocytes in ovine infarcts produced by six hours of complete ischemia followed by reperfusion. AB - BACKGROUND: In this study we tested the hypothesis that delayed reperfusion of ischemic myocardium-too late to save myocytes-attenuates infarct expansion and improves collagen synthesis. METHODS: The hypothesis was tested in a sheep model of anteroapical infarction that has no collateral blood flow to the area at risk. After coronary ligation or arterial occlusion for 1 or 6 hours, sheep had serial hemodynamic and quantitative echocardiographic studies before and after infarction and 2, 5, 8, and 12 weeks later. Hearts were examined by light and electron microscopy at 2 and 12 weeks; hydroxyproline and ratios of type I/III collagen were measured at 12 weeks. RESULTS: After coronary occlusion, left ventricular (LV) function progressively decreased and size increased to form an anteroapical aneurysm. After 1 hour of ischemia, neither resting LV size nor function changed; the infarct contained a midmyocardial scar between epicardial and endocardial muscle. After 6 hours of ischemia, LV function was significantly better than that in nonperfused sheep. Two weeks after 6 hours of ischemia, no viable myocytes were visible by light microscopy, but electron micrographs showed rare intact nucleated myocytes with scarce cytoplasmic myofibrils. At the 12th week epicardial and endocardial myocytes reappeared in the infarct. Infarct collagen type I/III ratios were 1.2 in reperfused groups and 0.7 in nonperfused sheep. CONCLUSIONS: Delayed reperfusion causes loss and subsequent reappearance of ovine epicardial myocytes, improves collagen type I/III ratios, and attenuates LV dilatation and loss of function. One hypothesis to explain the reappearance of myocytes is that reperfusion partially reverses an incomplete apoptotic process. PMID- 11426759 TI - Sarpogrelate inhibits serotonin-induced proliferation of porcine coronary artery smooth muscle cells: implications for long-term graft patency. AB - BACKGROUND: Serotonin can induce proliferation of vascular smooth muscle cells. We assessed the ability of a specific serotonin receptor antagonist, sarpogrelate, to inhibit proliferation of cultured porcine coronary artery smooth muscle cells. METHODS: Cell proliferation and mitotic activity were measured using 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide. To determine the effect of sarpogrelate on DNA (deoxyribonucleic acid), RNA (ribonucleic acid), and protein synthesis, radioactive incorporation of 3H-thymidine, 3H uridine, and 3H-phenylalanine, respectively, was used. Synthesis of DNA was also assessed by flow cytometry with propidium iodide as a fluorochrome. RESULTS: Serotonin, platelet-derived growth factor, endothelin, and angiotensin II all induced proliferation of porcine coronary artery smooth muscle cells. Sarpogrelate specifically inhibited the serotonin-induced cytokine trigger but did not influence platelet-derived growth factor-, endothelin-, or angiotensin II induced cell proliferation. Sarpogrelate inhibited the serotonin-induced increase in intracellular free ionized calcium concentration, prevented mitogen-activated protein kinase activation, and down-regulated expression of the protooncogenes c fos and c-jun. Sarpogrelate acted at the G1 phase of the cell cycle. CONCLUSIONS: These data suggest that sarpogrelate could be used as a therapeutic agent to inhibit serotonin-induced neointimal hyperplasia and improve patency of coronary artery bypass grafts. PMID- 11426760 TI - Vascular delay and intermittent stimulation: keys to successful latissimus dorsi muscle stimulation. AB - BACKGROUND: The goal of this study was to obtain physiologically significant increases in peak left ventricular (LV) systolic pressure and stroke volume with latissimus dorsi muscle (LDM) stimulation in cardiomyoplasty (CMP). We hypothesized that preserving LDM integrity by vascular delay and intermittent stimulation would significantly increase LDM cardiac assistance. METHODS: In 4 control dogs and 12 dogs that had undergone a vascular delay (VD) procedure, LV dysfunction was induced by intracoronary microsphere injections. Cardiomyoplasty surgery was performed 14 days later, followed by progressive LDM conditioning. In the control dogs and in 6 of the VD dogs, the LDM was stimulated 24 hours per day (VD plus constant stimulation [CS]). In the other 6 VD dogs, LDMs were stimulated on a daily schedule of 10 hours on and 14 hours off (VD plus interrupted stimulation [IS]). Latissimus dorsi muscle stimulated beats were compared with nonstimulated beats 9 weeks later. RESULTS: In the control dogs, LDM stimulation had minimal effects. In VD + CS and VD + IS, LDM stimulation increased peak LV pressure, stroke volume, stroke work, and stroke power (p < 0.05). However, these changes were greater in the VD + IS group, in which LDM stimulation increased peak aortic pressure by 17.6 +/- 1.7 mm Hg, peak LV pressure by 19.7 +/- 1.1 mm Hg, peak positive LV dp/dt by 398 +/- 144 mm Hg per second, stroke volume by 5.1 +/- 0.7 mL, stroke work by 10.9 +/- 0.9 gm.m, and stroke power by 122.7 +/- 11.6 gm.m per second (p < 0.05 compared with VD + CS). Quantitative morphometric analysis showed minimal LDM degeneration in the VD + IS group (7.5% +/- 1.1%), and VD + CS group (10.5% +/- 4.5%) compared with the control group (29.5% +/- 4.5%, p < 0.05). CONCLUSIONS: VD and IS considerably increased the LV assistance with LDM stimulation. Further studies of this combined approach to CMP should be planned. PMID- 11426761 TI - Double valve endocarditis. AB - BACKGROUND: There are little data concerning surgical outcomes in patients with native valve endocarditis affecting both the aortic and mitral valves. METHODS: From 1977 to 1998, 54 patients had simultaneous aortic and mitral valve grafting for native valve endocarditis. In 78%, mitral valve involvement was limited to the anterior leaflet, suggesting a jet lesion from the aortic valve. Surgical strategies included 31 valve repairs and valve replacement with mechanical (34), bioprosthetic (34), or allograft (9) prostheses. Three hundred twenty-five patient-years of follow-up were available for analysis (mean 6.0 +/- 4.8 years). RESULTS: There were no hospital deaths. Ten-year survival was 73%. Ten-year freedom from recurrent endocarditis was 84%, with risk peaking at 3 months, followed by a constant risk of 1.3%/yr. Choice of valvar procedure did not influence mortality or reinfection risk. CONCLUSIONS: The most common pattern of double valve infection was a jet lesion on the anterior mitral leaflet. Surgical treatment has late survival and freedom from reinfection similar to those of patients with single heart valve infection. PMID- 11426762 TI - Valvular heart operation in patients with previous mediastinal radiation therapy. AB - BACKGROUND: The outcome of valvular heart operations in patients with previous mediastinal radiation therapy was studied. METHODS: This is a single center retrospective study of 60 patients (37 females, 23 males) with a mean age of 62 +/- 15 years (28 to 88 years old) operated on from January 1976 to December 1998. Valvular heart operations performed included aortic valve replacements (n = 26), mitral valve procedures (n = 16), tricuspid valve procedures (n = 6), and multiple valve procedures (n = 12). A total of 264 clinical, hemodynamic, electrocardiographic and echocardiographic variables were analyzed. RESULTS: Total follow-up was 199 patient-years with a mean of 3.3 +/- 3.1 years and a range of 0 to 12.4 years old. Early mortality was 7 patients (12%). Early mortality in patients with constrictive pericarditis was 40% (4 of 10) compared with 6% (3 of 50) in patients without constrictive pericarditis. By univariate analysis, early mortality was associated with constrictive pericarditis (p = 0.011), reduced preoperative ejection fraction (p = 0.015), and longer cardiopulmonary bypass times (p = 0.037). A total of 14 patients (23%) required permanent pacemaker placement before (n = 7), during (n = 1), or early (n = 6) after valvular heart operations. There were 19 late deaths (malignancies, 7; heart failures, 5; other cardiac, 4; and other noncardiac, 3). Overall survival and freedom from late cardiac death and cardiac reoperation at 5 years for hospital survivors were 66% +/- 8%, 82% +/- 7%, and 93% +/- 4%, respectively. By univariate analysis, late cardiac death was associated with low ejection fraction (p = 0.002), New York Heart Association (NYHA) functional class IV (p = 0.004), preoperative congestive heart failure (p = 0.02), and preoperative atrial fibrillation (p = 0.038). Eighty-five percent of the discharged patients were in NYHA functional class I or II at follow-up. CONCLUSIONS: Early results of valve replacement after mediastinal radiation therapy were good except in the presence of constrictive pericarditis. Long-term outcome was limited by malignancy and heart failure. Early surgical intervention is recommended before the development of risk factors for late death, namely, severe symptoms, left ventricular dysfunction, and atrial fibrillation. PMID- 11426763 TI - Actuarial and actual analysis of surgical results: empirical validation. AB - This report validates the use of the Kaplan-Meier (actuarial) method of computing survival curves by comparing 12-year estimates published in 1978 with current assessments. It also contrasts cumulative incidence curves, referred to as "actual" analysis in the cardiac-related literature with Kaplan-Meier curves for thromboembolism and demonstrates that with the former estimate the percentage of events that will actually occur. PMID- 11426764 TI - Cardiac reoperation in the intensive care unit. AB - BACKGROUND: At our institution, cardiac reoperations are routinely performed in the cardiac intensive care unit, as opposed to taking these patients back to the operating room. Our hypothesis was that reoperation in a cardiac intensive care unit does not increase sternal infection rate. METHODS: A retrospective analysis was performed on 6,908 adult patients undergoing cardiac operation over a 9-year period. Excluding those in cardiac arrest, 340 (4.9%) patients underwent reoperation in the cardiac intensive care unit, of which 289 survived (85%). RESULTS: Of the 289 patients who survived reoperation in the intensive care unit, 6 developed wound infections that required operative debridement (2.1%), which was not significantly different from those patients not requiring reoperation (1.9%, 121 of 6,497, p = 0.70). Hospital charges for a 2-hour reoperation in the intensive care unit and operating room are approximately $1,972/patient and $5,832/patient, respectively. CONCLUSIONS: Reoperation in the intensive care unit does not increase wound infection rate compared to those without reoperation. Decreased charges, avoiding transport of potentially unstable patients, quicker time to intervention, and convenience are advantages of reoperation in an intensive care unit. PMID- 11426765 TI - Surgical experience with left ventricular free wall rupture. AB - BACKGROUND: Autopsy studies reveal that left ventricular free wall rupture (LVFWR) accounts for 7% to 24% of deaths after myocardial infarction. The condition occurs up to 10 times more often than papillary muscle or interventricular septal rupture. A high index of suspicion must be maintained to differentiate LVFWR from infarct extension, cardiogenic shock, pulmonary embolus, and even Dressler's syndrome. METHODS: Since 1980, we have operated on 18 patients with LVFWR. Fourteen patients had experienced "blow-out" rupture associated with cardiogenic shock. Four patients had "stuttering" ruptures, a less spectacular occurrence. Echocardiography was the most important diagnostic tool. Repair was performed, usually using infarctectomy and direct suture closure. RESULTS: Eleven patients (61%) died after operation, 4 patients as a result of rerupture 1 to 12 hours after operation. Recently, we have used a "patch/glue" technique to repair ruptures in 2 patients. We believe this technique is superior to direct suture closure in preventing rerupture. There have been 7 long-term survivors (39%) from 6 months to 15 years. CONCLUSIONS: Left ventricular free wall rupture is not always sudden and dramatic. Yet, the operating staff must be willing to race to the operating room even with the patient in full resuscitation. Echocardiography is the most sensitive and efficient diagnostic tool. All rupture sites should be aggressively repaired, possibly combining direct suture and patch/glue techniques. PMID- 11426766 TI - Closed chest hypothermic circulatory arrest for complex intracranial aneurysms. AB - BACKGROUND: Recent advances in techniques of cardiopulmonary bypass permitted hypothermic circulatory arrest (HCA) using groin cannulation with the chest closed (CC-HCA) and without direct access to the heart. Herein we describe our experience with this technique for complex intracranial aneurysms. METHODS: Between 1992 and 1999, 16 patients (4 men and 12 women) with a mean age of 52 years (range 32 to 61 years) with complex intracranial aneurysms underwent resection or clipping of their aneurysms at our institution using the technique of CC-HCA and groin cannulation. Groin access was obtained with 16F to 19F arterial and 18F to 20F venous cannulas placing the tips at the aortoiliac and atriocaval junctions, respectively. Patients were cooled to a nasopharyngeal temperature of 16 degrees C. RESULTS: Mean circulatory arrest time was 32 minutes. No patient required conversion to standard sternotomy and central cannulation. There were no intraoperative deaths. The 30-day hospital mortality was 2 of 16 patients (12%). Of the 14 surviving patients (88%), 1 developed bilateral third nerve palsy and another left hemiparesis that improved on follow up. Both were discharged to an extended care facility and continued to do well at home after discharge. Two patients developed deep venous thrombosis postoperatively and required anticoagulation. All patients continued to do well at a mean follow-up of 42 months. CONCLUSIONS: The less invasive technique of CC HCA through groin cannulation avoids complications associated with a sternotomy, is safe and is associated with little morbidity, reduced operative time, and early hospital discharge and rehabilitation. PMID- 11426767 TI - Prospective randomized neurocognitive and S-100 study of hypothermic circulatory arrest, retrograde brain perfusion, and antegrade brain perfusion for aortic arch operations. AB - BACKGROUND: To determine the optimal method of brain protection during deep hypothermic circulatory arrest (DHCA) for arch repair. METHODS: Of 139 potential aortic arch repairs (denominator), we randomized 30 patients to either DHCA alone (n = 10), DHCA plus retrograde brain perfusion (RBP) (n = 10), or antegrade perfusion (ANTE) (n = 10); a further 5 coronary bypass (CAB) patients were controls. Fifty-one neurocognitive subscores were obtained for each patient at each of four intervals: preoperatively, 3 to 6 days postoperatively, 2 to 3 weeks postoperatively, and 6 months postoperatively. Intraoperative and postoperative S 100 blood levels and electroencephalograms were also obtained. RESULTS: For the denominator, the 30-day and hospital survival rate was 97.8% (136 of 139) and the stroke rate 2.8% (4 of 139). For the randomized patients, the survival rate was 100% and no patient suffered a stroke or seizure. Circulatory arrest (CA) times were not different (DHCA: RBP:ANTE) for 11 total arch repairs (including 6 elephant trunk; mean, 41.4 minutes; standard deviation, 15). Hemiarch repairs (n = 17) were quickest with DHCA (mean 10.0 minutes; standard deviation, 3.6; p = 0.011) and longest with ANTE (mean 23.8 minutes; standard deviation, 10.28; p = 0.004). Of the patients, 96% had clinical neurocognitive impairment at 3 to 6 days, but by 2 to 3 weeks only 9% had a residual new deficit (1 DHCA, 1 RBP, 1 ANTE), and by 6 months these 3 patients had recovered. Comparison of postoperative mean scores showed the DHCA group did better than RBP patients in 5 of 7 significantly different (p < 0.05) scores and versus 9 of 9 ANTE patients. There were no S-100 level differences between CA groups, but levels were significantly higher versus the CAB controls, particularly at the end of bypass (p < 0.0001); however, these may have been influenced by other variables such as greater pump time, cardiotomy use, and postoperative autotransfusion. Circulatory arrest (p = 0.01) and pump time (p = 0.057) correlated with peak S-100 levels. CONCLUSIONS: The results of hypothermic arrest have improved; however, there is no neurocognitive advantage with RBP or ANTE. Nevertheless, retrograde brain perfusion may, in a larger study, potentially reduce the risk of strokes related to embolic material. S-100 levels may be artificial. In patients with severe atheroma or high risk for embolic strokes, we use a combination of retrograde and antegrade perfusion on a selective basis. PMID- 11426768 TI - S100beta correlates with neurologic complications after aortic operation using circulatory arrest. AB - BACKGROUND: Astrocyte protein S100beta is a potential serum marker for neurologic injury. The goals of this study were to determine whether elevated serum S100beta correlates with neurologic complications in patients requiring hypothermic circulatory arrest (HCA) during thoracic aortic repair, and to determine the impact of retrograde cerebral perfusion (RCP) on S100beta release in this setting. METHODS: Thirty-nine consecutive patients underwent thoracic aortic repairs during HCA; RCP was used in 25 patients. Serum S100beta was measured preoperatively, after cardiopulmonary bypass, and 24 hours postoperatively. RESULTS: Neurologic complications occurred in 3 patients (8%). These patients had higher postbypass S100beta levels (7.17 +/- 1.01 microg/L) than those without neurologic complications (3.63 +/- 2.31 microg/L, p = 0.013). Patients with S100beta levels of 6.0 microg/L or more had a higher incidence of neurologic complications (3 of 7, 43%) compared with those who had levels less than 6.0 microg/L (0 of 30, p = 0.005). Retrograde cerebral perfusion did not affect S100beta release. CONCLUSIONS: Serum S100beta levels of 6.0 microg/L or higher after HCA correlates with postoperative neurologic complications. Using serum S100beta as a marker for brain injury, RCP does not provide improved cerebral protection over HCA alone. PMID- 11426769 TI - Heparin-induced thrombocytopenia: bovine versus porcine heparin in cardiopulmonary bypass surgery. AB - BACKGROUND: Studies have demonstrated a high incidence of antibodies to heparin/platelet factor 4 complexes, the antigen in heparin-induced thrombocytopenia, in patients after cardiopulmonary bypass surgery. In many hospitals, beef lung heparin has been used historically for cardiopulmonary bypass, and there has been reluctance to change to porcine heparin despite concerns of an increased incidence of heparin-induced thrombocytopenia in patients receiving bovine heparin. METHODS: A prospective randomized trial comparing bovine and porcine heparin in cardiopulmonary bypass surgery was conducted. Presurgery and postsurgery heparin antibody formation was studied using the serotonin release assay and a heparin/platelet factor 4 enzyme-linked immunosorbent assay. RESULTS: Data available on 98 patients, randomized to receive either bovine or porcine heparin, revealed no significant difference in patient positivity by serotonin release assay (12% in both groups) or by the heparin/platelet factor 4 enzyme-linked immunosorbent assay (29% with porcine and 35% with bovine heparin) postoperatively. There were no significant differences between preoperative and postoperative platelet counts or thromboembolic complications. CONCLUSIONS: Our study does not support the belief that bovine heparin is more likely than porcine heparin to induce the development of antibodies to heparin/platelet factor 4. PMID- 11426770 TI - Lactate release during reperfusion predicts low cardiac output syndrome after coronary bypass surgery. AB - BACKGROUND: Cardioplegic arrest induces anaerobic myocardial metabolism with a net production of lactate from glycolysis. However, persistent lactate release during reperfusion suggests a delayed recovery of normal aerobic metabolism and may lead to depressed myocardial function necessitating inotropic or intraaortic balloon pump support (low output syndrome [LOS]). We examined the relation between perioperative myocardial metabolism and postoperative clinical outcomes in patients undergoing isolated coronary artery bypass surgery (CABG). METHODS: We reviewed 623 patients who were enrolled in clinical studies evaluating perioperative myocardial metabolism between 1983 and 1996. Arterial and coronary sinus blood samples were obtained intraoperatively to assess myocardial metabolism. Clinical data regarding patient demographics and postoperative outcomes were prospectively collected and entered into our institutional database. RESULTS: Low output syndrome developed in 36 patients (5.8%). Myocardial lactate release was higher in these patients compared with those who did not develop postoperative LOS. Advanced age and poor preoperative left ventricular function were independent predictors of lactate release during reperfusion. Persistent lactate release after 5 minutes of reperfusion was the only independent predictor of postoperative LOS in this low-risk population. CONCLUSIONS: Persistent lactate release during reperfusion occurs in a significant proportion of low-risk patients undergoing isolated CABG and is an independent predictor of postoperative low cardiac output syndrome. Persistent lactate release during reperfusion suggests a delayed recovery of aerobic myocardial metabolism and may be related to intraoperative misadventure or inadequate myocardial protection. Myocardial lactate release may be useful as an alternative end-point in clinical trials evaluating perioperative myocardial protection. PMID- 11426771 TI - Antiinflammatory effects of colforsin daropate hydrochloride, a novel water soluble forskolin derivative. AB - BACKGROUND: To evaluate the effects of colforsin daropate hydrochloride (colforsin), a water-soluble forskolin derivative, on hemodynamics and systemic inflammatory response after cardiopulmonary bypass, we conducted a prospective randomized study. METHODS: Twenty-nine patients undergoing coronary artery bypass grafting were randomized to receive either colforsin treatment (colforsin; n = 14) or no colforsin treatment (control; n = 15). Administration of colforsin (0.5 microg.kg(-1).min(-1)) was started after induction of anesthesia and was continued for 6 hours. Perioperative cytokine and cyclic adenosine monophosphate levels, hemodynamics, and respiratory function were measured serially. RESULTS: Marked positive inotropic and vasodilatory effects were observed in patients receiving colforsin. Interleukin 1beta, interleukin 6, and interleukin 8 levels after cardiopulmonary bypass were significantly (p < 0.05) lower in the colforsin group. Plasma levels of cyclic adenosine monophosphate increased significantly (p < 0.05) in the colforsin group, and the levels correlated inversely (r = -0.56, p = 0.002) with the respiratory index after cardiopulmonary bypass. CONCLUSIONS: Intraoperative administration of colforsin daropate hydrochloride had potent inotropic and vasodilatory activity and attenuated cytokine production and respiratory dysfunction after cardiopulmonary bypass. The results indicate that the technique can be a novel therapeutic strategy for the systemic inflammatory response associated with cardiopulmonary bypass. PMID- 11426772 TI - Optimal position of atrial epicardial leads for temporary pacing in infants after cardiac surgery. AB - BACKGROUND: The atrial epicardial wall of pediatric patients was topographically assessed to identify the optimal position for temporary atrial pacing. METHODS: Unipolar electrodes were fixed at the cephalic wall between the right and left atrial appendages, the interatrial groove, and the right atrial appendage of fifteen pediatric patients who underwent definitive surgical repair. The performance of the three electrodes in terms of pacing patterns and six combinations of bipolar pacing were evaluated in terms of the pacing threshold, P wave amplitude, slew rate, and lead impedance. RESULTS: Unipolar pacing of the right atrial appendage showed a significantly higher threshold than the other groups. A bipolar configuration of the cephalic atrial wall and interatrial groove had a significantly higher P-wave amplitude than groups without the electrode at the cephalic atrial wall, and a significantly higher slew rate than a unipolar configuration of the atrial appendage. CONCLUSIONS: Bipolar pacing with the negative electrode at the cephalic atrial wall and the indifferent electrode at the interatrial groove is the most efficient method for pediatric patients. PMID- 11426773 TI - Efficacy of single versus bilateral internal mammary artery grafting in women: a long-term study. AB - BACKGROUND: Coronary artery bypass grafting carries a higher operative mortality and less favorable long-term benefit in women than in men. Bilateral internal mammary artery grafting (BIMA) has been shown to yield excellent perioperative and long-term results in both women and men. However, controversy continues to exist as to the benefits of a second internal mammary artery graft in women. METHODS: A retrospective analysis was performed comparing 261 consecutive women from a single surgical practice receiving BIMA and supplemental vein grafts between January 1972 and October 1994 with a computer-matched cohort of 261 women receiving single internal mammary artery (SIMA) and vein grafts during the same period. Univariate analysis confirmed the homogeneity of the two groups based on nine preoperative variables. RESULTS: Operative mortality was comparable in the two groups, 3.8% (10 of 261 patients) in the SIMA and 3.4% (9 of 261 patients) in the BIMA group, with a markedly reduced mortality in both groups since 1990, 2.3% (2 of 86 patients) in the SIMA and 1.3% (1 of 78 patients) in the BIMA group. The mean number of distal grafts (2.78, SIMA; 3.14, BIMA), perfusion time (104 minutes, SIMA; 108 minutes, BIMA), and cross-clamp time (58 minutes, SIMA; 66 minutes, BIMA) were all comparable. There was no significant difference in the incidence of postoperative complications, including sternal wound infection. Patient follow-up ranged from 1 month to 27 years, with a mean of 10.0 years in the SIMA group and 9.1 years in the BIMA group. Clinical results were excellent, with 100% (136 of 136 patients) of the SIMA and 100% (167 of 167 patients) of the BIMA patients in Canadian Cardiovascular Society class I or II at follow-up. Rates of late myocardial infarction, percutaneous transluminal coronary angioplasty, and reoperation were similarly low in both groups: 3.7% (5 of 136 patients) versus 1.8% (3 of 166 patients), 5.4% (7 of 136 patients) versus 4.8% (8 of 166 patients), and 3.7% (5 of 136 patients) versus 1.8% (3 of 166 patients), for SIMA versus BIMA survivors, respectively. No significant difference was found in the long-term and event-free survival or in any of the eight subscales of the SF-36 quality of life survey for the two groups. CONCLUSIONS: Excellent short- and long-term results have been demonstrated with internal mammary artery grafting in women. However, the addition of a second internal mammary artery graft does not appear to confer any additional clinical benefits in a comparably matched cohort of patients. PMID- 11426774 TI - Off-pump reoperative coronary artery bypass grafting by thoracotomy: patient selection and operative technique. AB - BACKGROUND: Reoperative coronary artery bypass grafting (CABG) in patients with contraindications to sternotomy or cardiopulmonary bypass (CPB) presents a technical challenge. In this study we reviewed patient selection, operative technique, and early results in patients having redo-CABG to the circumflex artery system by a thoracotomy without CPB. METHODS: From January 1996 through December 1999, 21 patients with contraindications to conventional redo-CABG had target vessel revascularization off-pump by thoracotomy. A posterolateral thoracotomy approach was used. RESULTS: No patient required sternotomy or CPB. There was no hospital mortality. Postoperative cardiac morbidity included non-Q wave myocardial infarction (5%), need for intraaortic balloon pump support postoperatively (5%), and atrial fibrillation (5%). Two grafts were studied early and two were studied late (more than 6 months later). One venous graft was found to be occluded early. Survival at 2 years was 95%. Ninety percent of surviving patients were in New York Heart Association functional class I or II. CONCLUSIONS: This approach was associated with no mortality, low morbidity, and favorable early symptomatic improvement. This is the approach of choice in cases of reoperative CABG to the circumflex system when resternotomy or CPB are undesirable, and the culprit coronary vessels are accessible through a thoracotomy. PMID- 11426775 TI - Intraaortic balloon pump therapy facilitates posterior vessel off-pump coronary artery bypass grafting in high-risk patients. AB - BACKGROUND: Displacement of the heart to expose posterior vessels during coronary artery bypass grafting (CABG) without cardiopulmonary bypass (off-pump CABG, or OPCAB) may impair cardiac function. We used the intraaortic balloon pump (IABP) preoperatively to reduce operative risk and to facilitate posterior vessel OPCAB in high-risk patients with left main coronary artery disease (> 75% stenosis), intractable resting angina, postinfarction angina, left ventricular dysfunction (ejection fraction < 35%), or unstable angina. METHODS: One hundred and forty-two consecutive patients who underwent multivessel OPCAB including posterior vessel revascularization were studied prospectively. The patients were divided into group I (n = 57), which received preoperative or intraoperative IABP, and group II (n = 85), which did not receive IABP. In group I, there were 34 patients with left main coronary artery disease, 24 patients with intractable resting angina, 8 patients with left ventricular dysfunction, 5 patients with postinfarction angina, and 40 patients with unstable angina. Seven patients received intraoperative IABP support owing to hemodynamic instability during OPCAB. RESULTS: There was no operative mortality in group I and 1 death in group II. The average number of distal anastomoses was not different between group I and group II (3.4 +/- 0.9 versus 3.5 +/- 0.9, p = not significant). There were no significant differences in the number of posterior vessel anastomoses per patient. There were no differences in ventilator support time, length of stay in the intensive care unit, hospital stay, and morbidity between the two groups. There was one IABP-related complication in group I. CONCLUSIONS: IABP therapy facilitates posterior vessel OPCAB in high-risk patients, and surgical results are comparable with those in lower-risk patients. PMID- 11426776 TI - Myocardial revascularization with multiple arterial grafts: comparison between the radial artery and the right internal thoracic artery. AB - BACKGROUND: Bilateral internal thoracic artery (ITA) harvesting is significantly underused, whereas the radial artery is being used with increasing frequency. We have retrospectively analyzed perioperative and short-term outcomes of patients receiving a radial artery versus those receiving a right ITA as a second arterial graft. METHODS: Between February 1999 and May 2000, 250 patients underwent coronary artery bypass grafting using the radial artery (156 patients) or the right ITA (94 patients) in combination with the left ITA and, when required, the saphenous vein. RESULTS: There was a higher prevalence of risk factors in the radial artery group. More coronary artery bypass graftings (p < 0.001) were performed with the radial artery. Operative mortality was not different (p = not significant). In the right ITA group there was more bleeding (p < 0.001) and a longer hospital stay (p < 0.001). Mean follow-up was 8.1 +/- 3.9 months. The probability of survival was similar (p = not significant). CONCLUSIONS: The radial artery can extend the benefits of multiple arterial grafting to those patients who are usually excluded from bilateral ITA harvesting because of multiple risk factors. Perioperative and short-term results are good. PMID- 11426777 TI - Successful biventricular repair after initial Norwood operation for interrupted aortic arch with severe left ventricular outflow tract obstruction. AB - BACKGROUND: Management of newborns with interrupted aortic arch (IAA) remains challenging. Associated severe left ventricular outflow tract obstruction (LVOTO) have often led to increased mortality with neonatal biventricular repair. We review our experience with an alternative approach for this complex surgical problem. METHODS: From May 1991 to June 1999, 28 neonates were treated for IAA. Thirteen of 28 neonates (46%) had type B IAA, ventricular septal defect (VSD) and severe LVOTO (Z value -2 to -7; mean -5 +/- 1.7). Mean age was 8 days (3 to 23 days old) with average weight of 3.3 kg (2.4 to 4.2 kg). Eight of 13 (62%) had anomalous right subclavian artery. Ten of 13 (77%) had thymic aplasia and chromosome 22 region qll deletion. All 13 patients were treated initially with a modified Norwood procedure. RESULTS: There were no perioperative deaths. Complications included 2 patients with recurrent arch stenosis treated with balloon dilatation. Two patients had systemic arterial shunt revision. Follow-up ranged from 2 to 99 months old (mean 39 months). There were 2 late deaths unrelated to any operation. Nine of 12 patients had a second stage palliation consisting of a bidirectional Glenn shunt. Six patients went on to have biventricular repairs (3 Ross-Konno, 2 Rastelli, 1 VSD closure with LVOT resection). One patient had a modified Fontan operation and 5 patients are awaiting potential biventricular repair. CONCLUSIONS: Children with IAA and severe LVOTO may be managed by initial Norwood palliation with an excellent outcome likely. This initial "univentricular" approach has enabled eventual successful biventricular repair despite severe LVOTO. PMID- 11426778 TI - Arterial switch operation: factors impacting survival in the current era. AB - BACKGROUND: The arterial switch procedure has become the preferred procedure for the transposition of the great arteries (TGA) and Taussig-Bing anomaly. This analysis is intended to identify potential factors affecting survival in the current era. METHODS: From 1986 to 1999, 201 consecutive patients underwent an arterial switch operation for TGA or Taussig-Bing anomaly. Multivariate analysis of perioperative variables was performed for operative morbidity/ mortality. Patients were separated into two groups. Phase 1 (n = 29) included patients before mid-1989 who underwent an open coronary reimplantation technique. Phase 2 (n = 172) included the patients undergoing a technique of reimplanting coronary buttons after neoaortic reconstruction. RESULTS: The patient population included TGA with intact ventricular septum (58.7%, 118 of 201), with ventricular septal defect (31.3%, 61 of 201), and Taussig-Bing anomaly (10.0%, 22 of 201). Overall, early mortality was 9.5% (19 of 201) and there were five late deaths (2.7%). One month, 1-year, and 5-year actuarial survival rates were 90.4%, 87.9%, and 87.9%, respectively. Reoperation rate for late pulmonary stenosis was 2.7% (5 of 182). The freedom from reoperation at 3 and 5 years was 97.5% and 93.3%, respectively. In the analysis by time period, the operative mortality declined from 27.6% (8 of 29) to 6.4% (11 of 172) (p = 0.002). Risk factors for operative death were coronary artery patterns (usual vs retropulmonary left coronary artery, p = 0.009) in phase 1 and preoperative instability in phase 2 (p = 0.002). CONCLUSIONS: The arterial switch operation for TGA and Taussig-Bing anomaly has early low and late mortality and reoperation rates. Technical modifications in coronary reimplantation have minimized coronary artery pattern-related risks. PMID- 11426779 TI - Repair of anomalous left main coronary artery arising from the pulmonary artery in infants: long-term impact on the mitral valve. AB - BACKGROUND: Infants presenting with anomalous left coronary artery off the pulmonary artery (ALCAPA) are generally in heart failure and often have significant mitral valve regurgitation (MR). Although establishing a dual coronary circulation is the procedure of choice, there remains controversy as to how the mitral valve is handled. METHODS: We reviewed our experience with this lesion at St. Louis Children's Hospital. Over the past 15 years, 17 infants under 18 months of age have undergone repair, with all but one being treated with reimplantation of the left coronary artery into the aorta; the other underwent the Takeuchi procedure (intrapulmonary artery baffle) and was excluded from this evaluation. The average age and weight at operation were 0.5 +/- 0.3 years and 6.1 +/- 1.9 kg, respectively. All presented with varying degrees of heart failure and 9 patients also had either moderate or severe MR. RESULTS: There was one early and no late deaths after reimplantation of the left coronary artery. The left ventricular function postrepair improved from a preoperative shortening fraction of 0.19 +/- 0.09 to 0.34 +/- 0.08 (p < 0.01). Moderate or severe MR was present in 2 patients postoperatively, and both developed significant obstruction in the left coronary artery postoperatively as well. Both underwent mitral valve repair and revascularization of the left coronary artery. CONCLUSIONS: Excellent results can be obtained in the treatment of this very high-risk group of patients. Mitral valve repair is not generally necessary at the time of the initial operation. However, should MR recur or persist late, it may herald the presence of a coexistent, significant coronary stenosis. Cardiac catheterization should be performed to assess the patency of the left coronary artery before performing mitral valve surgery. PMID- 11426780 TI - Thrombus formation after the Fontan operation. AB - BACKGROUND: Thrombus formation is common after a Fontan operation. We investigated the frequency and location of thrombus in our population of children based on the type of Fontan operation performed. METHODS AND RESULTS: Between January 1987 and January 1999, 592 patients underwent echocardiography after Fontan operation and 52 (8.8%) had intracardiac thrombus. Median age at Fontan operation was 1.9 years (range 0.8 to 35.1). Freedom from thrombus was 92%, 90%, 84% and 82% at 1, 3, 8, and 10 years after Fontan operation, respectively. There was no difference in freedom from thrombus, based on type of operation (atriopulmonary vs. lateral tunnel) or presence of fenestration. Thrombus was detected in the systemic venous atrium in 26 (48%), in the pulmonary venous atrium in 22 (44%), in both atria in 1 (2%), in the hypoplastic left ventricular cavity in 2 (8%), and in the ligated pulmonary artery stump in 1 (2%). CONCLUSIONS: Thrombus formation occurs with equal frequency in all types of modifications and is seen in the pulmonary, as well as the systemic venous atria. Our study suggests that thrombus formation is inherent to the physiology after Fontan operation and is not related to the type of modification performed. PMID- 11426781 TI - One and a half ventricle repair with pulsatile bidirectional Glenn: results and guidelines for patient selection. AB - BACKGROUND: The guidelines for performing a one and a half ventricle repair with pulsatile bidirectional Glenn remains controversial. This retrospective report summarizes the experience of a single institution, with an attempt at providing an answer. METHODS: Fifty consecutive patients, aged 4 months to 42 years, underwent intracardiac repair along with a superior cavopulmonary connection. Twenty-seven of the patients had had previous surgical palliation. Repair consisted of patch closure of the ventricular septal defect (n = 25), tricuspid valve repair (n = 26), reconstruction of the right ventricular outflow tract (n = 34), transpulmonary annular patch (n = 34), right ventricle to pulmonary artery homograft conduit (n = 4), and concomitant repair of atrioventricular canal (n = 9). Ten patients were left with a fenestration in the atrial septum. RESULTS: There were six hospital deaths (12%) and two late deaths (4.5%). Forty-two survivors were followed from 8 months to 116 months. Eighty-eight percent are in functional class I. Actuarial survival at 97 months was 74%. CONCLUSIONS: Moderate right heart hypoplasia constitutes a safe anatomic category for a pulsatile bidirectional Glenn. It is advisable not to proceed with a one and a half ventricle repair if postoperative residual pulmonary artery hypertension is anticipated. Patients requiring an intricate intracardiac repair and those with concomitant right heart hypoplasia may be better suited for a Fontan type of repair to reduce the complexity of the procedure. PMID- 11426782 TI - Anti-HLA class I antibodies and pulmonary homograft function after the Ross procedure. AB - BACKGROUND: The Ross procedure provides excellent long-term results in the majority of patients. However, degeneration of the pulmonary homograft in some patients remains an unresolved problem that may be related to immunologic factors. Therefore, we studied the prevalence of antihuman leukocyte antigen (HLA) class I antibodies and echocardiographic results of homograft function at rest. METHODS: Forty-seven patients (37 men, 10 women; 47 +/- 15 years) were seen for echocardiography 1.1 to 63.9 months (median, 27 months) postoperatively. The presence of anti-HLA antibodies was tested against a panel of lymphocytes of 50 donors. RESULTS: Twenty-seven (57%) of the patients produced anti-HLA class I antibodies. No difference in the maximal or mean transhomograft pressure gradient, or in the frequency of homograft regurgitation according to the presence or absence of anti-HLA antibodies was found. However, the right ventricle was slightly but significantly larger in antibody-positive patients (26.3 +/- 4.2 versus 30.7 +/- 3.5 mm; p = 0.001). CONCLUSIONS: In the first years after the Ross procedure, we could not detect significant evidence of an association between anti-HLA class I antibodies and echocardiographic results of homograft function at rest in adults. PMID- 11426783 TI - Living-donor lobar lung transplantation for primary ciliary dyskinesia. AB - A ventilator-dependent patient with primary ciliary dyskinesia underwent successful living-donor lobar lung transplantation. The case was a 24-year-old woman who had developed recurrent lower respiratory infection and became ventilator-dependent due to severe bronchiectasis. Transmission electron microscopy of the resected bronchus demonstrated inner dynein arm deficiency. PMID- 11426784 TI - Rare clinical diagnosis of dendriform pulmonary ossification. AB - Dendriform pulmonary ossification, defined as "widespread heterotopic bone formation within the lungs," is a rare entity, which is usually diagnosed upon postmortem examination. The case of a 43-year-old man with dendriform pulmonary ossification is presented. In this patient's case, thoracoscopic biopsy was an excellent diagnostic modality for identifying dendriform pulmonary ossification. Although this entity has been reported rarely, with improved technology and survival we might expect its prevalence to increase. PMID- 11426785 TI - Delayed pulmonary hemorrhage 17 years after gunshot wound to the chest. AB - A 45-year-old male returned 17 years after a gunshot wound to the chest with intermittent hemoptysis that progressed to frank pulmonary hemorrhage. The complications of retained intrathoracic foreign bodies are briefly reviewed. PMID- 11426786 TI - Excision of an invasive thymoma: a cure for ulcerative colitis? AB - Thymoma is often associated with other diseases, including autoimmune diseases and endocrine disorders. Ulcerative colitis is a nonspecific inflammatory disease and is thought to be an immune-mediated disorder; however, coincidence of thymoma and ulcerative colitis is rare. We treated a 51-year-old woman with invasive thymoma and ulcerative colitis. After excision of the invasive thymoma with partial resections of pericardium, left lung, brachiocephalic vein, and left phrenic nerve, the ulcerative colitis was cured. PMID- 11426787 TI - Extrapleural resection of lung metastasis in a patient with dense pleural adhesions using VATS. AB - We report the case of a 78-year-old man with dense pleural adhesion who underwent a resection of a lung metastasis by video-assisted thoracic surgery (VATS) through an extrapleural approach. The approach for diagnosis and therapeutic wedge resection of a lung tumor by VATS is easier and safer than an intrapleural approach if the patient has dense pleural adhesions. PMID- 11426788 TI - Off-pump combined coronary artery bypass grafting and left upper lobectomy through left posterolateral thoracotomy. AB - A 61-year-old man with angina had a lesion in the left upper lobe of his lung on chest roentgenogram. Coronary angiography revealed a dominant circumflex lesion. Combined coronary artery bypass grafting (CABG) and left upper lobectomy was performed through left posterolateral thoracotomy without the use of cardiopulmonary bypass. Off-pump CABG abolishes the complications of cardiopulmonary bypass, while posterolateral thoracotomy provides a direct access to the circumflex vessels and is ideal for lung resection. PMID- 11426789 TI - Surgical treatment of annuloaortic ectasia with coronary aneurysm and fistula. AB - Annuloaortic ectasia associated with a giant aneurysm of the left coronary artery and a coronary artery fistula is extremely rare, and it is difficult to decide how to repair this complex lesion. The cause of the huge aneurysm of the left coronary artery in our patient was thought to be cystic medial necrosis, the coronary artery fistula, or both. The surgical management of this extremely rare pathological combination is described. PMID- 11426790 TI - Mitral valve replacement and endocavitary patch repair for a giant left ventricular pseudoaneurysm. AB - We present a case of a giant inferior left ventricular (LV) wall pseudoaneurysm. The patient had New York Heart Association class IV heart failure due to severe mitral valve regurgitation and poor LV function. Our operative approach included right thoracotomy, excision of the mitral valve, and patch repair of the pseudoaneurysm neck from inside of the dilated LV cavity followed by mitral valve replacement. Surgery was performed without aortic cross-clamping on a normothermic perfused beating heart. The patient had an uncomplicated cardiac recovery and is doing well 15 months after surgery. PMID- 11426791 TI - Organized thrombus of the tricuspid valve mimicking valvular tumor. AB - We report on a case of organized thrombus of the tricuspid valve mimicking a valve tumor. Preoperative transesophageal echocardiography showed the mass to have originated from the septal leaflet of the tricuspid valve. A pouch of the tricuspid valve and a ventricular septal defect were observed perioperatively, with the mass attached to the septal leaflet. Histologic examination revealed the mass to be an organized thrombus without tumor components. PMID- 11426792 TI - Emergent operation for percutaneous coronary rupture after abciximab administration. AB - A 72-year-old woman had rupture of a coronary artery during angioplasty in the setting of abciximab therapy. Prolonged reinflation of the balloon failed to produce closure of the perforated site. Emergency coronary ligation and bypass after abciximab administration was associated with excessive postoperative bleeding. PMID- 11426793 TI - Rupture of an aneurysm of a coronary arteriovenous fistula. AB - Coronary arteriovenous fistula is a relatively rare congenital malformation. The course of this malformation is generally benign, and rupture of the fistula is thought to be very rare. We report a case of rupture of an aneurysm of a coronary arteriovenous fistula that originated from the distal left circumflex coronary artery and drained into the right atrium; this rupture was successfully treated with an operation. PMID- 11426794 TI - Hypothermic arrest for descending aortic rupture in reoperative patients. AB - Establishing hypothermic bypass for repair of descending thoracic aortic rupture in reoperative patients presents unique challenges for the operative team. A higher risk of stroke, embolization, and malperfusion further increases overall morbidity and mortality. Traditional femoral arterial cannulation may not be the optimal route for bypass for these patients. We report two reoperative cases using the right subclavian artery for arterial inflow to avoid these problems. PMID- 11426795 TI - Atherosclerotic aneurysm in the circumflex retroesophageal right aortic arch. AB - A 63-year-old man had a distal arch aneurysm combined with a circumflex retroesophageal right aortic arch, which is a rare form of vascular ring. By using combined median sternotomy and left thoracotomy under deep hypothermia and retrograde cerebral perfusion, total arch replacement with a vascular graft was successful. We describe our surgical strategy for arch aneurysm associated with an uncommon congenital vascular malformation. PMID- 11426796 TI - Reconstruction of the left ventricle in a patient with cardiac hemangioma at the apex. AB - Cardiac hemangiomas in the left ventricle are extremely rare. A 34-year-old woman, without symptoms, with a diagnosis of cardiac tumor at the apex of the left ventricle was referred to us. The tumor was surgically resected, and the diagnosis was hemangioma. The Jatene technique, originally introduced for left ventricular aneurysmectomy was excellent for repair after resection of a cardiac tumor at the apex. PMID- 11426797 TI - Hydatid disease of the interventricular septum causing pericardial effusion. AB - Echinococcal infestation of the heart is uncommon. We report a case of a 35-year old man with an hydatid cyst located in distal interventricular septum. Clinical presentation was chest pain and urticaria. Diagnosis was made by echocardiography and magnetic resonance imaging. Surgical resection was performed; the cyst was punctured and its content was drained, hypertonic glucose solution was instilled for sterilization, and it was removed. The patient did well and remains asymptomatic. Diagnosis and ultimate surgical treatment of this disease prevented potentially lethal complications such as cyst rupture with embolic phenomena and anaphylactic shock. PMID- 11426798 TI - Dor's endoaneurysmorrhaphy in severe heart failure due to giant cell myocarditis. AB - Giant cell myocarditis is an unusual and frequently fatal form of myocarditis. A 37-year-old woman presented with resistant cardiac failure and left ventricular aneurysm. She underwent Dor's endoaneurymorrhaphy and was histopathologically proved to have giant cell myocarditis. She had significant improvement of symptoms and was alive 13 months after surgery. Dor's endoaneurysmorrhaphy may be a useful therapeutic modality in selected cases of giant cell myocarditis. PMID- 11426799 TI - Infection of a retained permanent epicardial pacemaker lead. AB - Infection of a retained permanent epicardial pacemaker lead rarely causes mediastinal infection. A 21-month-old boy who had undergone an arterial switch operation at day 6 of life presented with mediastinal infection 3 months after removal of the generator. Removal of the infected pacemaker leads with the inflammatory granuloma was performed under extracorporeal circulation. The mediastinal infection developed from the retained epicardial pacemaker lead infection. PMID- 11426800 TI - Staged unifocalization and anatomic repair in a patient with right isomerism. AB - Anatomic biventricular repair was successfully achieved subsequent to bilateral unifocalizations of the pulmonary blood supply using heterologous pericardial rolls in a patient with isomeric right appendages and major aortopulmonary collateral arteries. PMID- 11426801 TI - Bilateral absence of the superior vena cava. AB - Congenital anomalies of the superior vena cava are rare and often coexist with other cardiac abnormalities. In this report, we present a case of absent bilateral superior vena cava with no associated cardiac anomaly. Blood from the arms, head, and upper torso returned to the right atrium through the azygos vein and the inferior vena cava, mimicking chronic superior vena cava obstruction. PMID- 11426802 TI - Anomalous origin of innominate artery from right pulmonary artery in DiGeorge Syndrome. AB - We experienced a case of anomalous origin of innominate artery from right pulmonary artery (isolated innominate artery). This patient was a 2-month-old baby girl weighing 3.2 kg with DiGeorge syndrome, who was diagnosed with perimembranous ventricular septal defect, atrial septal defect, and patent ductus arteriosus. This type of anomaly is exceedingly rare. PMID- 11426803 TI - Anomalous left coronary artery arising from the right coronary cusp. PMID- 11426804 TI - Left ventricular volume reduction without ventriculectomy. AB - Partial left ventriculectomy (the Batista procedure) to achieve left ventricular volume reduction (LVVR) has been advocated as an alternative to cardiac transplantation in patients with end-stage dilated left ventricles. Here, we describe a new technique of LVVR that uses realignment of the papillary muscles, thus avoiding ventriculectomy, and report preliminary results. Eight patients (all male, mean age 49.3 [range 38 to 70] years) underwent LVVR between October 1998 and March 2000 as an adjunct to surgical coronary revascularization. Five were assessed with echocardiography and cardiopulmonary exercise testing before and after (mean follow-up time 267 [range 94 to 416] days) the operation. LVVR significantly improved left ventricular end-diastolic volume (254 +/- 32 to 218 +/- 36 mL, p = 0.03), left ventricular ejection fraction (20.14% +/- 1.36% to 31.28% +/- 2.32%, p = 0.007), and exercise duration (from 394 +/- 88 to 611 +/- 79 seconds, p = 0.03). A nonsignificant improvement in maximal oxygen consumption was also observed. This technique of LVVR is relatively simple to perform and is accomplished through a small apical cardiotomy. Preliminary results show an encouraging functional improvement following surgery. Future controlled studies are required to assess this novel technique further. PMID- 11426805 TI - A new modified Bentall procedure: the flanged technique. AB - Replacement of the aortic root with a composite graft containing a prosthetic mechanical valve is the preferred surgical procedure for tailoring the aortic root. A new composite graft is designed with an extension below the prosthetic valve. The flange of the graft is anastomosed to the aortic annulus with a continuous suture. The remainder of the procedure is performed in the usual fashion, with button coronary anastomoses. This method is an alternative to previously described Bentall procedures for all aortic root pathologies, especially in cases with small aortic root precluding root enlargement and in those with defects at the annular and subannular areas that require repair. PMID- 11426806 TI - Repair of atrial septal defect through a minimal right vertical infra-axillary thoracotomy in a beating heart. AB - We present a minimally invasive and cosmetic approach to repair atrial septal defect. Fifty-five atrial septal defects were successfully repaired through a minimal right vertical infra-axillary thoracotomy. Every repair was performed on the beating heart with continuous suction through aortic needle vents and with the head of the patient lowered. This approach can be used not only in adults but also in young girls. PMID- 11426807 TI - Repair of posterior coronary lacerations. AB - Posterior lacerations of the coronary arteries that occur during arteriotomy should be repaired if significant. We describe a modification of the standard technique to repair posterior coronary artery lacerations. This technique avoids distortion and narrowing of the coronary artery. PMID- 11426808 TI - Epicardial atrial pacemaker lead placement after multiple cardiac operations. AB - We describe a simple technique for the implantation of left atrial epicardial pacing leads in children with congenital heart disease who have undergone multiple operations. The pulmonary veins are exposed to reveal the pulmonary venous to atrial confluence using a left thoracotomy. A pacemaker lead is secured to the posterior left atrium inferior to the lower pulmonary vein. This approach provides a reliable site for atrial lead placement without the need for extensive dissection. PMID- 11426809 TI - Blunt tracheobronchial injuries: treatment and outcomes. AB - BACKGROUND: Tracheobronchial injury is a recognized, yet uncommon, result of blunt trauma to the thorax. Often the diagnosis and treatment are delayed, resulting in attempted surgical repair months or even years after the injury. This report is an extensive review of the literature on tracheobronchial ruptures that examines outcomes and their association with the time from injury to diagnosis. METHODS: We reviewed all patients with blunt tracheobronchial injuries published in the literature to determine the anatomic location of the injury, mechanism of the injury, time until diagnosis and treatment, and outcome. Only patients with blunt intrathoracic tracheobronchial traumas were included. RESULTS: We identified 265 patients reported between 1873 and 1996. Motor vehicle accidents were the most frequent mechanism of injury (59%). The overall mortality among reported patients has declined from 36% before 1950 to 9% since 1970. The injury occurred within 2 cm of the carina in 76% of patients, and 43% occurred within the first 2 cm of the right main bronchus. The proximity of the injury to the carina had no detectable effect on mortality. Injuries on the right side were treated sooner but were associated with a higher mortality than left-sided injuries. No association was detected between delay in treatment and successful repair of the injury; ninety percent of patients undergoing treatment more than 1 year after injury were repaired successfully. CONCLUSIONS: This review of patients with blunt tracheobronchial injuries represents the largest cohort studied to date. These data suggest an ability to repair tracheobronchial injuries successfully many months after they occur. We are also able to assess the mortality associated with the location and side of injury, examine the time from injury until diagnosis and treatment, and evaluate treatment outcome. PMID- 11426810 TI - Therapeutic implications of inflammation in atherosclerotic cardiovascular disease. AB - Atherosclerosis represents a spectrum of pathologic lesions with diverse clinical sequelae. In this review, we build upon the paradigm that arteriosclerosis represents an inflammatory disease. By examining mechanisms involved in the response to vascular injury, we can more effectively implement targeted therapy aimed at halting or regressing arteriosclerosis. PMID- 11426811 TI - The trachea. PMID- 11426812 TI - As originally published in 1993: right latissimus dorsi cardiomyoplasty augments left ventricular systolic performance. Updated in 2001. PMID- 11426813 TI - As originally published in 1994: dynamic descending thoracic aortomyoplasty: comparison with intraaortic balloon pump in a model of heart failure. Updated in 2001. PMID- 11426814 TI - Improved long-term survival seen after lung volume reduction surgery compared to continued medical therapy for emphysema. PMID- 11426815 TI - Diaphragm flap for routine prophylactic reinforcement of bronchial stump after pneumonectomy. PMID- 11426816 TI - Pleural tent after upper lobectomy: routine or selective? PMID- 11426817 TI - Chordal replacement or repair. PMID- 11426818 TI - Cardiac surgery in octogenarians with poor lung function. PMID- 11426819 TI - S100 release as an indicator of cerebral damage. PMID- 11426820 TI - Oral hypoglycemic agents in coronary artery bypass grafting. PMID- 11426821 TI - Off-pump coronary reoperations via left thoracotomy. PMID- 11426822 TI - Impact of myocardial infarction on the outcome after partial left ventriculectomy. PMID- 11426823 TI - Of cells and architecture: new approaches to old criteria in mycosis fungoides. PMID- 11426824 TI - Histologic criteria for the diagnosis of mycosis fungoides: proposal for a grading system to standardize pathology reporting. AB - BACKGROUND: The histological diagnosis of early lesions of mycosis fungoides (MF) is often difficult for dermatopathologists and prior studies have shown a low agreement rate among pathologists. An important reason for such difficulty may be the lack of specific histological criteria. METHODS: We tested a new method to interpret and report biopsies suspicious for MF. The method is based on a grading system reflecting the pathologist's degree of diagnostic certainty. A panel of four pathologists independently assessed a set of 50 biopsies suspicious for MF first without (Phase I) and subsequently with specific histological criteria (Phase II). A third Phase was carried out after a training session, using a new set of cases with corresponding immunophenotyping and gene rearrangement analysis. Weighted and unweighted kappa statistics were used to assess inter- and intra-pathologist variation. RESULTS: The consensus rate among pathologists improved from 48% in Phase I to 76% in Phase III. Overall precision weighted kappas increased from 0.630 in Phase I to 0.854 in Phase III, indicating excellent inter-pathologist agreement by Phase III. There was a significant association between the presence of an abnormal phenotype and/or T-cell clonality and a higher diagnostic score. CONCLUSIONS: The use of uniform criteria and training sessions can substantially improve the consensus rate among pathologists in the diagnosis of MF. PMID- 11426825 TI - Expression of Basigin in human fetal, infantile and adult skin and in basal cell carcinoma. AB - BACKGROUND: Basigin is a glycosylated transmembrane protein belonging to the immunoglobulin superfamily and is thought to be associated with cell development and differentiation. We investigated the relation between Basigin expression and epidermal development in this study. METHODS: Basigin expression was immunohistochemically investigated during organogenesis of human skin and in human basal cell carcinoma (BCC). RESULTS: Human fetal skin showed negative staining at 10 weeks of gestation. At 20 weeks, the cytoplasm and membranes of adnexal germ and hair follicular cells were strongly positive, while epidermal basal cells showed weakly positive staining. After birth, basal cells, suprabasal cells, anagen hair follicular cells and eccrine glandular cells showed positive staining. Membranes of basal cells expressed more Basigin compared to other cell components. Basigin was not detectable in granular cells and telogen hair follicular cells. Sixteen of 30 BCCs were entirely negative for Basigin. However, cells at budding areas of tumor masses were positive in 14 of the 30 BCCs. CONCLUSIONS: These findings suggest that 1) Basigin is associated with epidermal proliferation and differentiation, 2) most parts of BCCs might be derived from early fetal epidermal basal cells, and 3) that a part, only the budding area of BCCs, has the characteristics of epithelial germ cells. PMID- 11426826 TI - E-cadherin expression in wound healing of mouse skin. AB - BACKGROUND: E-cadherin has been studied extensively in other systems but little attention has been paid to its role in wound healing. We investigated E-cadherin expression in epithelial wound healing in vivo by focusing on the migrating cells in the epithelial tongue and the mitotic cells in the non-injured side apart from the original wound edge. METHODS: Round full-thickness excisional wounds (6 mm in diameter) and full-thickness incisional wounds were prepared dorsally in mice. On various days after the operation, E-cadherin expression was examined by immunohistochemical staining using a monoclonal antibody specific for E-cadherin. RESULTS: In both models, the level of E-cadherin expression did not decrease on the 1st postoperative (P.O.) day. After the 2nd P.O. day, E-cadherin expression decreased in cells at a site 500 microm apart from the original wound edge. After the 3rd P.O. day, decreased expression was also observed in cells at the top and in the basal layer of the epithelial tongue. This decreased expression continued for 1 or 2 days after the meeting of the epithelial tongue. There was no significant difference in the expression of E-cadherin between two models. CONCLUSIONS: The results suggested that E-cadherin expression decreases in epithelial cells. This decrease may depend on the activity of migration and mitosis. In addition, the change was similar in both the excisional and incisional wounds. PMID- 11426827 TI - The clonality of tumor-infiltrating lymphocytes in African Kaposi's sarcoma. AB - BACKGROUND: African Kaposi's sarcoma (KS) lesions contain human herpesvirus-8 (HHV-8) and Epstein-Barr virus (EBV), both of which are associated with various types of non-Hodgkin's lymphomas and are known to produce several factors suspected of lymphomagenic potential. The aim of this study was to evaluate tumor infiltrating lymphocytes for the evidence of clonal expansion in African KS. METHODS: We used polymerase chain reaction (PCR)-based assays to determine the clonality of tumor-infiltrating lymphocytes in African KS lesions and compared the results to similar studies of patient-matched uninvolved skin and peripheral blood. RESULTS: T cells were polyclonal in all samples tested. Peripheral blood B cells were also polyclonal; however, a minority of lesional and uninvolved skin samples exhibited evidence of restricted B-cell clonality. Correlation with immunohistological analysis revealed that this clonal B-cell restriction was secondary to the sparse nature of lesional B cells rather than their clonal overgrowth. CONCLUSIONS: We conclude that, despite the putative lymphomagenic potential of HHV-8 and EBV and their co-existence in African KS lesions, tumor infiltrating lymphocytes in these cases do not show evidence of clonal expansion that might be an early manifestation of lymphoma. Nevertheless, these studies are a case in point that sparse lymphoid subpopulations in lesional and uninvolved extranodal tissues can give rise to restricted clonal patterns that must be interpreted carefully to avoid the misdiagnosis of occult lymphoma. PMID- 11426828 TI - Human dermal pericytes express 3G5 ganglioside--a new approach for microvessel histology in the skin. AB - BACKGROUND: Pericytes cover the abluminal surface of microvessels and play an important role in capillary regulation and pathology. Studies on pericytes have been hindered by the lack of specific markers with which to facilitate microscopic identification of this cell type. Expression of the cell surface 3G5 ganglioside antigen has been reported in cultured retinal and cardiac pericytes. The objective of this study was to determine the usefulness of monoclonal antibody 3G5 as a pericyte marker in human skin. METHODS: Cryosections of 21 skin biopsies were examined by direct fluorescence technique with anti-3G5, anti-von Willebrand factor, anti-alpha-smooth muscle actin or DNA fluorochrome. RESULTS: In human dermis, 3G5 expression is limited to pericytes discriminating this cell type from all other cells including smooth muscle cells, myofibroblasts and myoepithelial cells. We found a pericyte: endothelial cell ratio of 1:12.4 (+/ 7.1), and a difference of alpha-smooth muscle actin expression between the subpapillary plexus and the microvessels of the Stratum reticulare. CONCLUSIONS: 3G5 mAB is an excellent and so far the only reported tool for identification of dermal pericytes by fluorescent light microscopy. Moreover, this is the first report of the application of 3G5 technique to the microvasculature in tissue sections at the light microscopic level. PMID- 11426829 TI - Restrictive dermopathy: report of a case and review of the literature. AB - BACKGROUND: Restrictive dermopathy is a rare autosomal recessive skin disorder that is fatal in the neonatal period. Clinical and pathologic findings are distinctive and allow for a specific diagnosis in most cases. METHODS: We present a case of an affected infant and a review of the previously reported cases in the literature. RESULTS: The infant had thick shiny skin with reduced compliance and multiple spontaneous linear splits. Additional findings included an abnormal facies with a distinctive small, round and open mouth, low set ears, small nose, widely spaced sutures, flexion contractures of the extremities, and poorly expanded lungs. The infant expired 65 h after birth. Histologic findings of the skin at autopsy included a relatively unremarkable epidermis, a flat dermal epidermal junction (absent rete ridges), an overall thinned dermis with hypoplastic appendage structures, a dense fibrotic reticular dermis with collagen parallel to the epidermis, a sharp subcutaneous margin, and an abnormally thick layer of subcutaneous adipose tissue. Electron microscopic findings included dense dermal patches of collagen and fibroblasts with abundant endoplasmic reticulum and unusually small tonofilaments. Review of previously reported cases reveals strikingly consistent findings. CONCLUSIONS: This rare condition illustrates that abnormal cutaneous development may produce fetal hypokinesia, leading to profound effects on intrauterine growth and development. The autosomal recessive pattern of inheritance and morphologic changes of the skin and skeletal system in this disorder suggest that a structural protein or enzyme defect, perhaps of collagen metabolism, may underlie the pathogenesis. PMID- 11426830 TI - Can we reliably use markers of cell cycle regulation (e.g. Ki-67, p21, p27, p53) to differentiate between benign and malignant skin tumors? PMID- 11426831 TI - Characterisation of agonist binding on human 5-HT2C receptor isoforms. AB - The 5-HT2C receptor is expressed in different isoforms as a result of mRNA editing. Both INI (unedited) and VSV (a fully edited version) isoforms are abundant in rat brain. The VSV isoform lacks the high affinity recognition site for 5-HT, which may be caused by low efficiency coupling to G-proteins. In this study we have investigated the pharmacology of the agonist binding site of these two isoforms of the 5-HT2C receptor. The VSV isoform was expressed in Chinese hamster ovary cells (CHO) and the INI isoform in both Chinese hamster ovary cells and human embryonic kidney cells (HEK-293). Saturation analysis using [3H]5-HT revealed high and low affinity recognition sites on the INI isoform in both cell types whilst the VSV isoform did not have the high affinity binding site for [3H]5-HT. Displacement studies were undertaken using [3H]5-HT to label the receptors. In these studies the affinity of agonists (5-HT, Ro600175 ((S)-2-(6 Chloro-5-fluoroindol-1-yl)-1-methylethylamine), MK212 (6-Chloro-2-(piperazinyl) pyrazine), mCPP (1-(m-chlorophenyl)-piperazine), TfMPP (N-(m trifluoromethylphenyl)piperazine), DOI (1-(2,5-Dimethoxy-4-iodophenyl)-2 aminopropane), DOB (1-(4-bromo-2,5-dimethoxyphenyl)-2-aminopropane) and 8OH-DPAT (8-hydroxy-2-(di-N-propylamino)tetralin) was higher at the INI isoform, whilst antagonist affinity (ketanserin and mesulergine) did not change between the two receptor isoforms. There were no differences between the INI isoform expressed in the CHO and HEK-293. This suggests that the INI isoform of the 5-HT2C receptor is pharmacologically similar to the VSV form of the 5-HT2C receptor but that it couples more efficiently to G-proteins. PMID- 11426832 TI - Characterization of organic anion transport inhibitors using cells stably expressing human organic anion transporters. AB - The organic anion transport system is involved in the tubular excretion of various clinically important drugs. The purpose of this study was to characterize the effects of various organic anion transport inhibitors on organic anion transport using proximal tubule cells stably expressing human organic anion transporter 1 (human-OAT1) and human-OAT3, which are localized to the basolateral membrane of the proximal tubule. Organic anion transport inhibitors including betamipron, cilastatin, KW-3902 (8-(noradamantan-3-yl)-1,3-dipropylxanthine) and probenecid significantly inhibited human-OAT1- and human-OAT3-mediated organic anion uptake in a dose-dependent manner. Kinetic analyses revealed that these inhibitions were competitive. The Ki values of betamipron, cilastatin, KW-3902 and probencid for human-OAT1 were 23.6, 1470, 7.82 and 12.1 microM, whereas those for human-OAT3 were 48.3, 231, 3.70 and 9.0 microM. These results suggest that betamipron and probenecid could inhibit both human-OAT1- and human-OAT3-mediated organic anion transport in vivo, whereas cilastatin could inhibit only human-OAT3 mediated one. In contrast, KW-3902 did not exert the effects of significance, whereas KW-3902 was the most potent. PMID- 11426833 TI - FK506 inhibits Cl- secretion in airway epithelium via calcineurin-independent mechanism. AB - FK506 (tacrolimus)-binding protein (FKBP) is associated with intracellular Ca2+ release channel and modulates its function. To elucidate the effect of FK506 on Ca2+ dynamics and Ca2+-mediated Cl- secretion in airway epithelium, we studied intracellular Ca2+ ([Ca2+]i) concentration and Cl(-)-dependent short-circuit current (Isc), in cultured bovine tracheal epithelial cells. Addition of ATP induced an increase in [Ca2+]i, and this response was dose dependently inhibited by FK506. Rapamycin, which binds FKBP with high affinity, likewise inhibited the [Ca2+]i rise, but cyclosporin A, a specific calcineurin inhibitor, did not. In Cl secretion studies using Ussing chamber, ATP increased Ca2+-mediated Isc in amiloride-treated cells, an effect that was inhibited by FK506 and rapamycin but not by cyclosporin A. Therefore, FK506 inhibits Ca2+ mobilization in airway epithelium via FKBP but not calcineurin-dependent mechanism, which may result in the suppression of Ca2+-activated Cl- secretion. PMID- 11426834 TI - Effect of alisol B acetate, a plant triterpene, on apoptosis in vascular smooth muscle cells and lymphocytes. AB - Glucocorticoid-induced apoptosis is a well-recognized physiological regulator of T-cell number and function. Alisol B acetate, a triterpene from Alisma Plantago aquatica, has a glucocorticoid-like structure, and may have a similar function like glucocorticoid-induced apoptosis in both vascular smooth muscle cell line (A7r5) and human acute lymphoblastic leukemia cell line (CEM cells). For exploring its mechanism, mitochondria membrane potential and apoptosis-related gene expression were discussed. Alisol B (10(-6)-10(-4) M) inhibited serum stimulated DNA synthesis in a concentration-dependent manner (IC50) = 4.0 +/- 0.8 x 10(-6) M in A7r5 and 2.1 +/- 1.2 x 10(-6) M in CEM cells). The cell viability was reduced at 10(-4) M of alisol B. Similar results were seen in dexamethasone treatment (a synthetic glucocorticoid, 10(-6) M, 48 h). Apoptosis was induced after the cells were exposed to 10(-5)-10(-4) M alisol B or 10(-6) M dexamethasone for 48 h. The mitochondrial membrane potential (delta psi(m)) was significantly reduced after the alisol B treatment, indicating that the mitochondria might play a role in the alisol B induced cell apoptosis. Alisol B (10(-5)-10(-4) M) increased the levels of c-myc and bax mRNA and proteins, but not on the anti-apoptotic proto-oncogene, bcl-2, in A7r5 and CEM cells. In contrast, dexamethasone (10(-6) M) treatment only caused significant increase in c-myc mRNA levels. These results suggest that the increased ratio of Bax/Bcl-2 and the decreased mitochondrial membrane potential might be involved in the mechanisms of alisol B-induced cell apoptosis. PMID- 11426835 TI - Melanin potentiates daunorubicin-induced inhibition of collagen biosynthesis in human skin fibroblasts. AB - One of the recognized side effects of antineoplastic anthracyclines is poor wound healing, resulting from an impairment of collagen biosynthesis. The most affected tissue is skin. The mechanism underlying the tissue specificity of the side effects of anthracyclines has not been established. In view of the fact that a number of pharmacologic agents are known to form complexes with melanin and melanins are abundant constituents of the skin, we determined whether daunorubicin interacts with melanin and how this process affects collagen biosynthesis in cultured human skin fibroblasts. Results indicated that daunorubicin forms complexes with melanin. Scatchard analysis showed that the binding of daunorubicin to melanin was heterogeneous, suggesting the presence of two classes of independent binding sites with K1 = 1.83 x 10(5) M(-1) and K2 = 5.52 x 10(3) M(-1). The number of strong binding sites was calculated as n1 = 0.158 micromol/mg of melanin and the number of weak binding sites as n2 = 0.255 micromol/mg of melanin. We have suggested that prolidase, an enzyme involved in collagen metabolism, may be one of the targets for anthracycline-induced inhibition of collagen synthesis. We found that daunorubicin induced inhibition of prolidase activity (IC50 = 10 microM), collagen biosynthesis (IC50 = 70 microM) and DNA biosynthesis (IC50= 10 microM) in human skin fibroblasts. Melanin (100 microg/ml) by itself produced about 25% inhibition of DNA synthesis and prolidase activity but it had no effect on collagen biosynthesis in cultured fibroblasts. However, the addition of melanin (100 microg/ml) to daunorubicin treated cells (at IC50 concentration) augmented the inhibitory action of daunorubicin on collagen and DNA biosynthesis without having any effect on prolidase activity. The same effect was achieved when the cells were treated with daunorubicin at one-fourth of the IC50 given at 0, 6, 12 and 18 h during a 24-h incubation. The data suggest that the melanin-induced augmentation of the inhibitory effects of daunorubicin on collagen and DNA biosynthesis may result from: (i) accumulation of the drug in the extracellular matrix, (ii) gradual dissociation of the complex, and (iii) constant action of the released drug on cell metabolism. The phenomenon may explain the potential mechanism for the organ specificity of daunorubicin-induced poor wound healing in patients administered this drug. PMID- 11426836 TI - The neuroprotective activity of the glycine receptor antagonist GV150526: an in vivo study by magnetic resonance imaging. AB - The neuroprotective activity of GV150526 (3-[2-(Phenylaminocarbonyl)ethenyl]-4,6 dichloroindole-2-carboxylic acid sodium salt), a selective glycine receptor antagonist of the NMDA receptor, has been evaluated by magnetic resonance imaging (MRI) in a rat model of middle cerebral artery occlusion. The aim of the work was to evaluate, using an in vivo method, whether GV150526 was able to reduce the extent of ischemic brain damage when administered both before and after (6 h) middle cerebral artery occlusion. GV150526 was administered at a dose of 3 mg/kg i.v. T2-weighted (T2W) and diffusion weighted (DW) images were acquired at 6, 24 and 144 h after the establishment of the cerebral ischemia. Substantial neuroprotection was demonstrated at all investigated time points when GV150526 was administered before the ischemic insult. The ischemic volume was reduced by 84% and 72%, compared to control values, when measured from T2W and DW images, acquired 24 h after middle cerebral artery occlusion. Administration of the same dose of GV150526, 6 h post-ischemia, also resulted in a significant (p < 0.05) neuroprotection. The ischemic volume was reduced by 48% from control values when measured from T2W images and by 45% when measured from DW images. No significant difference was found between volumes of brain ischemia obtained by either MRI or triphenyltetrazolium chloride staining. These data confirm the potential neuroprotective activity of the glycine receptor antagonist GV150526 when administered either before or up to 6 h after ischemia. PMID- 11426837 TI - Delta9-tetrahydrocannabinol enhances cortical and hippocampal acetylcholine release in vivo: a microdialysis study. AB - The intravenous administration of synthetic cannabinoid agonists was recently shown to dose dependently increase acetylcholine release from the rat prefrontal cortex and hippocampus (Eur. J. Pharmacol. 401 (2000) 179]. We report here that the active ingredient of cannabis preparations, delta9-tetrahydrocannabinol, administered at 10, 37.5, 75 and 150 microg/kg, dose dependently stimulated acetylcholine release from rat prefrontal cortex and hippocampus estimated by means of in vivo brain microdialysis with vertical concentric probes. At these doses, delta9-tetrahydrocannabinol induced behavioural stimulation. The administration of the CB1 receptor antagonist, ([N-(piperidin-1-yl)-5-(4 chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3carboxamide]HCl) SR 141716A (200 microg/kg i.p.) significantly reduced the effect of delta9 tetrahydrocannabinol (75 microg/kg i.v.) on acetylcholine release from rat prefrontal cortex and hippocampus. PMID- 11426838 TI - Conformationally restricted analogs of BD1008 and an antisense oligodeoxynucleotide targeting sigma1 receptors produce anti-cocaine effects in mice. AB - Cocaine's ability to interact with sigma receptors suggests that these proteins mediate some of its behavioral effects. Therefore, three novel sigma receptor ligands with antagonist activity were evaluated in Swiss Webster mice: BD1018 (3S 1-[2-(3,4-dichlorophenyl)ethyl]-1,4-diazabicyclo[4.3.0]nonane), BD1063 (1-[2-(3,4 dichlorophenyl)ethyl]-4-methylpiperazine), and LR132 (1R,2S-(+)-cis-N-[2-(3,4 dichlorophenyl)ethyl]-2-(1-pyrrolidinyl)cyclohexylamine). Competition binding assays demonstrated that all three compounds have high affinities for sigma1 receptors. The three compounds vary in their affinities for sigma2 receptors and exhibit negligible affinities for dopamine, opioid, GABA(A) and NMDA receptors. In behavioral studies, pre-treatment of mice with BD1018, BD1063, or LR132 significantly attenuated cocaine-induced convulsions and lethality. Moreover, post-treatment with LR132 prevented cocaine-induced lethality in a significant proportion of animals. In contrast to the protection provided by the putative antagonists, the well-characterized sigma receptor agonist di-o-tolylguanidine (DTG) and the novel sigma receptor agonist BD1031 (3R-1-[2-(3,4 dichlorophenyl)ethyl]-1,4-diazabicyclo[4.3.0]nonane) each worsened the behavioral toxicity of cocaine. At doses where alone, they produced no significant effects on locomotion, BD1018, BD1063 and LR132 significantly attenuated the locomotor stimulatory effects of cocaine. To further validate the hypothesis that the anti cocaine effects of the novel ligands involved antagonism of sigma receptors, an antisense oligodeoxynucleotide against sigma1 receptors was also shown to significantly attenuate the convulsive and locomotor stimulatory effects of cocaine. Together, the data suggests that functional antagonism of sigma receptors is capable of attenuating a number of cocaine-induced behaviors. PMID- 11426839 TI - Antinociceptive action of amlodipine blocking N-type Ca2+ channels at the primary afferent neurons in mice. AB - We investigated the antinociceptive action of amlodipine, a dihydropyridine derivative, which acts on both L- and N-type voltage-dependent Ca2+ channels (VDCCs), in mice. Intrathecal injection of amlodipine (300 nmol/kg) significantly shortened the licking time in the late phase of a formalin test, while no effect was found with another dihydropyridine derivative, nicardipine (300 nmol/kg). Cilnidipine and omega-conotoxin GVIA also showed marked analgesic effects under the same experimental conditions. Transcripts of alpha1A, alpha1B, alpha1E, alpha1F, alpha1H, beta3, and beta4 subunits were detected by polymerase-chain reaction (PCR) in the dorsal root ganglion, suggesting the existence of a variety of voltage-dependent Ca2+ channels. Electrophysiological experiments showed that amlodipine and cilnidipine inhibit N-type currents in the dorsal root ganglion cells. These results suggest that amlodipine, cilnidipine, and omega-conotoxin GVIA exert their antinociceptive actions by blocking N-type Ca2+ channels in the primary nociceptive afferent fibers. Blocking of the Ca2+ channels results in attenuation of synaptic transmission of nociceptive neurons. Furthermore, it is suggested that some N-type Ca2+ channel blockers might have therapeutic potential as analgesics when applied directly into the subarachnoidal space. PMID- 11426840 TI - Role of brain arachidonic acid cascade on central CRF1 receptor-mediated activation of sympatho-adrenomedullary outflow in rats. AB - The present experiments were designed to characterize the mechanisms involved in the corticotropin releasing factor (CRF)-induced activation of central sympatho adrenomedullary outflow in rats. Intracerebroventricularly (i.c.v.) administered CRF and urocortin (0.5, 1.5 and 3.0 nmol/animal) effectively and dose-dependently elevated plasma levels of adrenaline and noradrenaline, and the effect of urocortin was almost the same as that of CRF. The elevation of catecholamines induced by CRF and urocortin (1.5 nmol/animal) was reduced by CP-154,526(butyl ethyl-(2,5-dimethyl-7-(2,4,6trimethylphenyl)-7H-pyrrolo [2,3-d] pyrimidin-4 yl]amine), a selective CRF1 receptor antagonist, in a dose dependent manner (1.2 and/or 2.4 micromol/animal, i.c.v.), and abolished by indomethacin (1.2 micromol/animal, i.c.v.), an inhibitor of cyclooxygenase. Furegrelate (1.8 micromol/animal, i.c.v.), an inhibitor of thromboxane A2 synthase, abolished the CRF-induced elevation of adrenaline, but had no effect on the evoked release of noradrenaline. These results suggest that activation of brain CRF1 receptor facilitates the central sympathetic and adrenomedullary outflow in distinct central pathways in rats; brain thromboxane A2 is involved in the central adrenomedullary outflow; an active metabolite of arachidonic acid other than thromboxane A2 (probably prostaglandin E2) may be involved in the central sympathetic outflow. PMID- 11426841 TI - Antinociceptive activity of the endogenous fatty acid amide, palmitylethanolamide. AB - The endogenous fatty acid ethanolamide, palmitylethanolamide, alleviated, in a dose-dependent manner, pain behaviors elicited in mice by injections of formalin (5%, intraplantar), acetic acid (0.6%, 0.5 ml per animal, intraperitoneal, i.p.), kaolin (2.5 mg per animal, i.p.), and magnesium sulfate (120 mg per kg, i.p.). The antinociceptive effects of palmitylethanolamide were prevented by the cannabinoid CB2 receptor antagonist SR144528 [N-([1s]-endo-1.3.3 trimethylbicyclo[2.3.1]heptan-2-yl)-5-(4-chloro-3-methylphenyl)-1-(4 methylbenzyl)-pyrazole-3-carboxamide], not by the cannabinoid CB1 receptor antagonist SR141716A [N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4 dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide x HCl]. By contrast, palmitylethanolamide had no effect on capsaicin-evoked pain behavior or thermal nociception. The endogenous cannabinoid, anandamide (arachidonylethanolamide), alleviated nociception in all tests (formalin, acetic acid, kaolin, magnesium sulfate, capsaicin and hot plate). These effects were prevented by the cannabinoid CB1 receptor antagonist SR141716A, not the cannabinoid CB2 receptor antagonist SR141716A. Additional fatty acid ethanolamides (oleylethanolamide, myristylethanolamide, palmitoleylethanolamide, palmitelaidylethanolamide) had little or no effect on formalin-evoked pain behavior, and were not investigated in other pain models. These results support the hypothesis that endogenous palmitylethanolamide participates in the intrinsic control of pain initiation. They also suggest that the putative receptor site activated by palmitylethanolamide may provide a novel target for peripherally acting analgesic drugs. PMID- 11426842 TI - Effects of repeated GBR 12909 administration on brain stimulation reward. AB - Male rats were trained at three separate currents to bar press for intracranial self-stimulation. On days 1 and 15, all subjects were given 1-(2-bis(4 fluorophenyl)-methoxy)-ethyl-4-(3-phenylpropyl) piperazine, also known as GBR 12909 (10 mg/kg, i.p.), prior to test session. Between these days, the paired Chronic-before group was injected (every other day) with GBR 12909 prior to intracranial self-stimulation, while unpaired, Chronic-after group was given the drug just after the end of the session. A third group (Control) received saline injections (i.p.) 20 min following the session. Although GBR 12909 was found to be reward enhancing, neither sensitization nor tolerance developed to the rewarding and performance/motor effects regardless of the injection regimen. In addition, the rewarding effects of intracranial self-stimulation were found to be independent of both current and environment-specific pairing. The present data obtained for GBR 12909 agree with previous observations of the effects of repeated administration of drugs of abuse on intracranial self-stimulation. PMID- 11426843 TI - Critical role of the endogenous cannabinoid system in mouse pup suckling and growth. AB - Delta9-tetrahydrocannabinol, the active principle in marijuana, is a cannabinoid receptor agonist. Both the crude drug and delta9-tetrahydrocannabinol have been used as appetite promoters. The endogenous cannabinoid, arachidonoyl ethanolamide (anandamide), likewise a cannabinoid receptor agonist, has been shown to have the same effect. In contrast, the cannabinoid CB1 receptor antagonist N-(piperidin-1 yl)-5(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1-H-pyrazole-3-carboxamide (SR141716A) reduces food intake. Here, we report that administration of SR141716A to newly born mouse pups (either a single administration on postnatal day 1, or daily for a week as of postnatal day 2) had a devastating effect on milk ingestion and growth. The first 24 h after birth appeared the most critical for the growth stunting effect of SR141716A. Death followed within 4-8 days. Co administration of delta9-tetrahydrocannabinol almost fully reversed the effect of the antagonist in the week-long regimen. Co-administration of 2-arachidonoyl glycerol, an endocannabinoid, with 2-palmitoyl glycerol and 2-linoleoyl glycerol, which enhance 2-arachidonoyl glycerol potency, resulted in a significant delay in mortality rates caused by the antagonist. We conclude that the endocannabinoid system plays a vital role in milk suckling, and hence in growth and development during the early stages of mouse life. PMID- 11426844 TI - Functional and biochemical evidence for capsaicin-induced neural endothelin release in isolated working rat heart. AB - In isolated working rat heart, capsaicin elicited a concentration-dependent constriction of coronary arteries accompanied by decline of all cardiac parameters recorded (heart rate, coronary and aortic flow, left ventricular developed pressure, and first derivative of left ventricular developed pressure). The following evidence suggests that capsaicin-induced changes are mediated by endothelin of neural origin: (1) the capsaicin (10 nM)-evoked decrease in coronary flow resulting in deterioration of cardiac functions was mimicked by endothelin (0.1 nM); (2) the selective endothelin ET(A) receptor antagonist, cyclo (D-alpha-aspartyl-L-propyl-D-valyl-L-leucyl-D-tryptophyl) (1 microM), abolished the cardiac effects provoked by capsaicin (10 nM); (3) reduction of extracellular Ca2+ concentration from 2.4 to 1.2 or 0.6 mM inhibited the cardiac effects of capsaicin (10 nM) but not those induced by endothelin (0.1 nM); (4) perfusion of the heart with 0.1% (v/v) Triton X-100 damaged the endothelium and reversed the enhancement of coronary flow evoked by bethanechol (1 microM), decreased the basal flow, but was without effect on capsaicin-induced coronary constriction; (5) in response to capsaicin challenge (10-100 nM), the endothelin concentration measured in coronary effluent by means of radioimmunoassay increased up to sevenfold but remained unchanged in the presence of 0.6 mM Ca2+; (6) no reduction of coronary flow was induced by capsaicin (100 nM) applied to the heart of rats which were desensitised by capsaicin (150 mg/kg). It is concluded that, in the rat heart, capsaicin acting on VR1 capsaicin receptors elicits a release of endothelin from the sensory nerve terminals. PMID- 11426845 TI - Differential effects of cocaine on the positive inotropic effect of noradrenaline mediated by alpha1- and beta-adrenoceptors in failing human myocardium. AB - Electrically driven (1 Hz) ventricular trabeculae from explanted failing human myocardium were indirectly examined for the localization of the alpha1 adrenoceptor population and the beta-adrenoceptor population in relation to sympathetic nerve endings. We examined the influence of neuronal uptake blockade by cocaine upon the horizontal position of the concentration-response curves for the inotropic effects exerted by noradrenaline in the presence and absence of appropriate adrenoceptor antagonists. Cocaine shifted the concentration-response curve for alpha1-adrenoceptor stimulation, but not that for beta-adrenoceptor stimulation, to lower concentrations of noradrenaline in a parallel manner. The concentration-response curve for combined adrenoceptor stimulation was shifted by cocaine to lower concentrations of noradrenaline in a nonparallel manner. In explanted allograft heart, cocaine had no effect upon the position of the concentration-response curve to alpha1-adrenoceptor stimulation. The data indicate that in the explanted native hearts the alpha1-adrenoceptor population is located close to or within the synaptic cleft, while the beta-adrenoceptor population remaining in the failing myocardium is located more distantly to the neuronal release sites. PMID- 11426846 TI - Adrenergic nerves mediate acetylcholine-induced endothelium-independent vasodilation in the rat mesenteric resistance artery. AB - Mechanisms underlying acetylcholine-induced endothelium-independent vasodilation were studied in the rat mesenteric vascular bed isolated from Wistar rats. In preparations without endothelium, and contracted by perfusion with Krebs solution containing methoxamine (2-7 microM), perfusion of acetylcholine (1-100 microM) for 1 min produced a concentration-dependent vasodilation. Denervation of denuded preparations by cold storage (4 degrees C for 72 h) abolished the acetylcholine induced vasodilation; 10 and 100 nM atropine abolished 1 and 10 microM acetylcholine-induced vasodilation, but it inhibited only 20% of vasodilation by 100 microM acetylcholine. The acetylcholine-induced atropine-resistant vasodilation was inhibited by 10 and 100 microM hexamethonium, 5 microM guanethidine, 50 microM bretylium, in vitro 6-hydroxydopamine (2 mM for 20 min, twice), 1 microM capsaicin and 0.5 microM calcitonin gene-related peptide (CGRP) (8-37) (CGRP receptor antagonist). These findings suggest that the acetylcholine induced endothelium-independent nicotinic vasodilation requires the presence of intact adrenergic nerves, and is mediated by endogenous CGRP released from CGRP containing nerves. PMID- 11426847 TI - Improved survival with simendan after experimental myocardial infarction in rats. AB - This study compared the effects of simendan, a calcium sensitizer, with those of milrinone and enalapril on survival of rats with healed myocardial infarction. Seven days after ligation-induced myocardial infarction, the rats were randomized to control, milrinone, enalapril, or simendan groups. All compounds were administered via the drinking water for 312 days, at which time there was 80% mortality in the control group--the study's primary endpoint. The infarct sizes were similar across all groups. At endpoint, the mortality rates were: 63% (milrinone), 56% (enalapril) and 53% (simendan); the risk reductions were 25% (P = 0.04 vs. control) and 28% (P = 0.02 vs. control) with enalapril and simendan, respectively. Milrinone had no statistically significant effect on the survival rate. These findings suggest that, like enalapril, simendan improved survival in rats with healed myocardial infarction. PMID- 11426848 TI - Pertussis toxin-sensitive and -insensitive mechanisms of alpha1-adrenoceptor mediated inotropic responses in rat heart. AB - In rat left ventricular papillary muscle, phenylephrine, an alpha1-adrenoceptor agonist, induced a triphasic inotropic response; an initial transient, small, positive inotropic effect followed by a transient chloroethylclonidine-sensitive negative inotropic effect and a sustained 2-(2,6 dimethoxyphenoxyethyl)aminomethyl-1,4-benzodioxane (WB4101)-sensitive positive inotropic effect. Treatment with pertussis toxin for 2 days significantly inhibited only the transient negative inotropic effect without changing the sustained positive inotropic effect. This treatment also prevented the acetylcholine (1 microM)-induced negative inotropic effect. Further, phenylephrine-induced transient negative inotropic effect was attenuated in the presence of ouabain. These results suggest that pertussis toxin-sensitive or insensitive G-protein may be responsible for alpha1-adrenoceptor subtype-mediated negative inotropic effect or positive inotropic effect, respectively, in which the transient negative inotropic effect was produced via the stimulation of Na+, K+ pump, presumably through pertussis toxin-sensitive G-protein-dependent pathway. PMID- 11426849 TI - Effects of fenoterol on beta-adrenoceptor and muscarinic M2 receptor function in bovine tracheal smooth muscle. AB - Prolonged (18 h) incubation of isolated bovine tracheal smooth muscle with the beta2-adrenoceptor agonist fenoterol (10 microM) induced desensitization of isoprenaline-induced adenylyl cyclase activity in bovine tracheal smooth muscle membranes, characterized by a 25% decrease in maximal effect (Emax) (P < 0.05), while the sensitivity to the agonist (pEC50) was unchanged. The Emax value of isoprenaline-induced smooth muscle relaxation of submaximal methacholine-induced contractile tones was similarly reduced by about 25% (P < 0.001), while the pEC50 value was diminished by 1.0 log unit (P < 0.001). As determined by 30 microM gallamine-induced muscarinic M2 receptor antagonism and pertussis toxin-induced inactivation of G(i alpha), muscarinic M2 receptor-mediated functional antagonism did not play a role in isoprenaline-induced relaxation of bovine tracheal smooth muscle contracted by methacholine, both in control and in 18-h fenoterol-treated tissue. In line with these observations, we found no enhanced muscarinic M2 receptor-mediated inhibition of 1 microM forskolin-stimulated adenylyl cyclase activity after 18-h fenoterol treatment. These data indicate that 18-h fenoterol treatment of bovine tracheal smooth muscle induces beta2-adrenoceptor desensitization and reduced functional antagonism of methacholine-induced contraction by beta-adrenoceptor agonists, without a change of muscarinic M2 receptor function. PMID- 11426850 TI - Tryptase-induced airway microvascular leakage in guinea pigs: involvement of tachykinins and leukotrienes. AB - Tryptase, a serine protease synthesized by and stored in mast cells, is implicated as an important mediator in the pathogenesis of airway inflammation. In this study, tryptase was evaluated for its ability to induce microvascular leakage into the airways of guinea pigs. Dose- and time-dependent increases in airway microvascular leakage were produced by intratracheal tryptase (0.3-3 microg). Intratracheal tryptase (3-30 microg) had no effect on airway tone as measured by pulmonary insufflation pressure. Tryptase-induced airway microvascular leakage was partially blocked by the tachykinin NK1 receptor antagonist CP 99994 [(+)-(2S,3S)-3-(2-methoxybenzylamino)-2-phenylpiperidine] and an inhibitor of leukotriene formation SCH 37224 (1-(1,2-dihydro-4-hydroxy-2-oxo-1 phenyl-1,8-naphthyridin-2-yl)pyrrolidinium, hydroxide inner salt). Neither CP 99994 nor SCH 37224 inhibited tryptase proteolytic activity in-vitro. Pretreatment of guinea pigs with histamine H1 receptor antagonists or a tachykinin NK2 receptor antagonist had no affect on the airway microvascular leakage induced by tryptase. It is speculated that tryptase may be important in the pathogenesis of airway inflammation, particularly in disorders that involve increased airway microvascular leakage such as asthma. PMID- 11426851 TI - Diacerein suppresses the increase in plasma nitric oxide in rat adjuvant-induced arthritis. AB - We investigated the effects of rhein, an active metabolite of diacerein, on the interleukin-1alpha-stimulated production of nitric oxide (NO) in rabbit articular chondrocytes, and the effects of diacerein on NO production in rat adjuvant induced arthritis. At doses of 10 and 30 microM, rhein significantly inhibited the interleukin-1alpha-stimulated NO production in chondrocytes. In the rat adjuvant-induced arthritis model, diacerein was administered for 21 days, starting at the time of adjuvant injection. Paw swelling and plasma NO level were measured in order to assess the effect of diacerein on arthritis and NO biosynthesis in the whole body. At doses of 30 and 100 mg/kg/day, diacerein significantly suppressed the development of adjuvant-induced arthritis and the increase in plasma NO. These results suggest that the inhibitory effect of diacerein on rat adjuvant-induced arthritis is partly related to its reduction of the NO production induced by adjuvant-induced arthritis. PMID- 11426852 TI - Xylem colonization of tomato by Azorhizobium caulinodans ORS571. AB - Tomato seedlings growing aseptically in Murashige and Skoog Medium were inoculated with Azorhizobium caulinodans ORS571 (pXLGD4), carrying the lacZ reporter gene. By microscopic analyses of inoculated tomato roots, it has been demonstrated that the xylem of tomato roots can be colonized by Azorhizobium. We discuss whether this colonization of the xylem of tomato roots by diazotrophic azorhizobia might provide a suitable niche for endophytic nitrogen fixation. PMID- 11426853 TI - Identification of yeasts isolated from poultry meat. AB - In an assessment of the potential role of yeasts in the spoilage of poultry meat, the population and species diversity of yeasts were determined on 50 commercial raw and processed chicken and turkey product samples. Initial populations (log10 cfu/g) ranged from less than 0.1 to 2.9, and increased by the expiration date to 0.4 to 5.1, respectively. 145 of 152 isolates were identified as belonging to 12 species. Yarrowia lipolytica and Candida zeylanoides were predominant, accounting for 39% and 26% of the isolates, respectively. Six different species of basidiomycetous yeasts were determined representing 24% of the isolates. The ability of the predominant yeast species to grow at refrigeration temperatures and their proteolytic and lipolytic activies suggest that yeasts may play a more significant role than previously recognised in the spoilage of poultry products. PMID- 11426854 TI - Interaction between mitochondria derived from incompatible black Aspergillus isolates. AB - As black Aspergillus isolates are highly heterokaryon-incompatible mitochondrial transmissions were performed by protoplast fusion. Donor strains with oligomycin resistant mitochondria and sensitive recipient partners of various A. japonicus isolates were applied and the progeny were selected for oligomycin resistance and for recipient nuclear phenotype. These strains basically inherited the mitochondrial DNA of the donor strain, which might remain unchanged (substituted progeny) or might be modified by specific sequences of the recipient mtDNAs (recombinant progeny). Different mobile elements characteristic of the recipient parents were exclusively responsible for the development of the feature of recombinant mtDNAs. Substituted progeny were either stable wild-type-like strains as a result of compatible co-operation between donor mitochondria and recipient nuclei, or aconidial strains with a reduced fitness, exhibiting a certain instability. The latter type was probably due to the less compatible communication between nuclear and extrachromosomal genetic systems originating from different parents. These progeny were able to undergo some developmental (segregation) processes during subsequent cultivation, resulting in a stable, wild-type phenotype which possessed a new type of mtDNA resembling that of the acceptor parents. PMID- 11426855 TI - Surface structures of new and lesser known species of thermobifida as revealed by scanning electron microscopy. AB - Surface structures of representatives of the genus Thermobifida were examined by scanning electron microscopy. Spores formed at the tips of multibranched sporophores initially resembled short sausages; then, upon maturation, they gradually built up their typical ovoid shape. Characteristic differences were observed between T. cellulolytica strain TB108 and T. fusca strains TM51. The spores of TB108 were larger (0.8x 1.3 microm) than those of TM51 (0.6 x 1.1 microm) in consequence of the more thickened outer squamous layer. When Thermobifida strains were grown on cellulose as sole carbon source, the mycelium was found to coil around the cellulose crystals and multiple protuberances emerged, resulting in a scabrous appearance to the mycelial surface. The presence of these cellulosome-like structures yielded a 24.5% surface enlargement of the scabrous mycelium as compared with the smooth one. The cellulosome emergence pattern paralleled the proportional increase in free endoglucanase activity measured during the culturing of these actinomycetes in the presence of cellulose. PMID- 11426856 TI - Characterization of the extracellular enzyme systems of Trichoderma viride AH124. AB - A mycoparasitic Trichoderma viride strain was investigated for the production of extracellular enzymes important in antagonism, by using natural and chromogenic substrates. Some of these enzymes, such as beta-1,3-glucanases, and low levels of proteases were produced constitutively. Under inductive conditions, the measurable activities of beta-1,3-glucanase, protease and aspecific chitinase increased, while for the proteases and beta-1,3-glucanases, the levels depended on both the nitrogen and the carbon source. Gel filtration chromatography revealed at least 4 beta-1,3-glucanases, 6 proteases, 2 beta-glucosidases and 1 beta-1,4-N-acetyl-glucosaminidase izoenzyme under inductive conditions. PMID- 11426857 TI - Ethanol-induced cell aggregation (flocculation) and its physiological background in Schizosaccharomyces pombe rive 4-2-1. AB - Cell aggregation (flocculation) of the yeast Schizosaccharomyces pombe strain RIVE 4-2-1 developed in glucose-containing medium, but only in the presence of ethanol. Cell surface proteins which participated in cell to cell interactions were characterised by the susceptibility of flocculation to different proteolytic enzymes, heat treatment, denaturing and thiol compounds and by the inhibition of flocculation by sugars and derivatives. It was shown that a galactose-specific lectin was involved in this new type of flocculation. PMID- 11426858 TI - Post-antibiotic and synergic effects of fluoroquinolones and ceftazidime in combination against Pseudomonas strains. AB - Infectious complications are still the leading cause of morbidity and mortality in seriously ill patients [1-3]. To combat the resistance of (mainly Gram negative, non-fermentative) bacteria, e.g. Pseudomonas aeruginosa, to a wide spectrum of antibiotics, drug combination therapy has been widely adopted as standard clinical practice since the late 1990s. beta-Lactam combinations are not optimal and the potential of nephrotoxicity and ototoxicity from aminoglycosides has caused clinicians to evaluate new possibilities, such as combinations of fluoroquinolones and beta-lactams. We examine here the synergic and post antibiotic effects (PAEs) of ciprofloxacin, ofloxacin and pefloxacin-ceftazidime combinations against 6 clinical Pseudomonas isolates. The fluoroquinolone ceftazidime combinations were not only synergic against Comamonas (P.) testosteroni, but had double the PAEs of the two drugs alone. The ciprofloxacin ceftazidime combination had a longer PAE against P. aeruginosa isolate 1 than ciprofloxacin alone. The combinations, however, did not have longer PAEs than those of the single drugs against the other 5 P. aeruginosa isolates. PMID- 11426859 TI - Fungal genotype controls mutualism and sex in Brachypodium sylvaticum infected by epichloe sylvatica. AB - The fungal endophyte Epichloe sylvatica (Clavicipitaceae, Ascomycota) may obligatorily infect the woodland grass Brachypodium sylvaticum, on which it can display two alternative modes of reproduction. During the sexual cycle, external stromata suppress host flowering and production of seed (choke disease), whereas in the asexual cycle the fungus remains asymptomatic and transmits vertically by seeds. Variation in the reproductive system thus determines whether the symbiosis is mutualistic or parasitic. In order to assess the relative effects of each genotype on fungal reproduction, we used naturally infected seed families of B. sylvaticum and experimentally infected plants with different combinations of plant and fungal genotypes. The results of one experiment suggested a maternal effect of the host association on the choke rate in the offspring, while the results of a second experiment clearly indicated that the fungal genotype determines stroma formation and thus the mode of reproduction. Since sexual reproduction of the fungus is closely tied with disease expression on the host, the fungal genotype may also be responsible for whether an endophyte association is beneficial or pathogenic. We discuss the results in the light of current theories about the evolution of mutualism and the maintenance of sex. PMID- 11426860 TI - Cell density-correlated induction of pyruvate decarboxylase under aerobic conditions in the yeast Pichia stipitis. AB - During the aerobic batch cultivation of P. stipitis CBS 5776 with glucose, pyruvate decarboxylase was activated in a cell number-correlated manner. Activation started when a cell number between 7 x 10(7) and x 10(8) cells ml(-1) was reached and the enzyme activity increased during further cultivation. This induction might have been triggered either by an unknown quorum sensing system or by a shortage of cytoplasmic acetyl-CoA. PMID- 11426861 TI - Occurrence of metronidazole and imipenem resistance among Bacteroides fragilis group clinical isolates in Hungary. AB - During the period between 1987 and 1997, various surveillances of the antibiotic resistance of B. fragilis group isolates revealed that practically all the isolates tested were susceptible to imipenem, metronidazole and chloramphenicol; very few isolates (2.5%) exhibited resistance to amoxicillin/clavulanic acid. However, similarly as in some southern European countries, the percentages of the isolates that were resistant to ampicillin, tetracycline and clindamycin were high throughout this period, and the resistance to cefoxitin increased from 6% to 16%. In 2000, isolates with intermediate or high resistance to imipenem and isolates with increased MICs to metronidazole were emerging among the clinical isolates of B. fragilis. The presence of the cfiA gene was demonstrated by PCR in 7 of 242 isolates (2.9%); 2 of them with high MICs to carbapenems harboured the IS942 element immediately upstream of the resistance genes. In the 2 B. fragilis isolates with increased MICs to metronidazole, the nim gene could be detected by PCR. The IS1186 element was found in these isolates upregulating the metronidazole resistance gene. PMID- 11426862 TI - Cell-wall composition and polysaccharide synthase activity changes following photoinduction in Trichoderma viride. AB - The differentiation and metabolism in the soil-borne saprophytic deuteromycete Trichoderma viride are subject to control by light. We have investigated the effect of illumination of mycelia on the activities of cell-wall synthesizing enzymes beta-1,3-glucan synthase and chitin synthase and on the composition of cell alls. After 10 min illumination of dark-grown mycelia with white fluorescent light at 600 1x, a gradual rise in specific activity of membrane-bound enzymes beta-1,3-glucan synthase by about 130% and a decrease in specific activity of chitin synthase by about 50% in relation to the dark control were observed. The changes in enzyme activities were caused by de novo synthesis of corresponding polysaccharide synthases(s) and/or their regulatory components since they were not observed when protein synthesis was blocked with 50 microg/ml cycloheximide. The content of beta-1,3-glucan in the cell walls of illuminated mycelia has increased by 35-50% in comparison to the dark control while the content of chitin remained practically unchanged. PMID- 11426863 TI - Variation of isoenzyme and RAPD patterns in Candida albicans morphological mutants with altered colony ultrastructure. AB - Molecular typing methods were applied to characterize four stable morphological mutants [1] isolated from a UV-induced unstable mutant colony of Candida albicans. The wild-type strain (ATCC 64385), the intermediate unstable mutant and its four morphologically altered derivatives revealed the same electrophoretic karyotypes. Of the five isoenzymes tested (catalase, malate dehydrogenase, glutamate dehydrogenase, acid phosphatase and 3-glucosidase), glutamate dehydrogenase displayed a different enzyme pattern (with an extra band of lower mobility) in the morphological mutants. In contrast, the random amplification DNA polymorphism patterns of the mutant strains differed in all cases from that of the parental strain. Different primers revealed various degrees of DNA polymorphism; one of them (OPC-8) proved to be useful for differentiation between all examined strains. Differences in genetic alterations between spontaneous and induced mutants, and the applicability of different molecular markers to analyse the consequences of induced mutagenesis in C. albicans are discussed. PMID- 11426864 TI - Effects of double-stranded RNA viruses on the reproduction of Phaffia rhodozyma. AB - DsRNA viruses were transferred from a virus-containing strain to a virus-free strain of Phaffia rhodozyma by protoplast fusion. The resulting new strain carried all three types of dsRNA of the virus-containing strain and had the electrophoretic karyotype of the virus-free strain. The effects of the dsRNA viruses on the host fitness were checked by following the asexual and the sexual reproductivity. The results demonstrated that viruses have no effect on the growth rate during the lag and log phases of the vegetative reproduction, but the maximum cell numbers in the stationary phase differ significantly. Inconclusive results were obtained as concerns the effects of viruses on the sexual reproduction. PMID- 11426865 TI - Production and regeneration of protoplasts from Gremmeniella abietina and Ascocalyx abietis. AB - This work was undertaken to develop a system of protoplast isolation and regeneration for G. abietina and A. abietis that could be of use for the genetic manipulation of both species. Nuclear staining was performed to assess the nuclear conditions of the protoplasts. Of the 19 enzyme complexes studied, only 10 were found to have some lytic effect on either Gremmeniella, A. abietis, or both. Only snail enzyme (from Szeged University), Novozym 234, lytic enzyme L1, or mixtures of snail enzyme and Novozym 234 produced satisfactory yields of protoplasts. Regeneration of protoplasts was observed on complete and on minimal medium, and occurred from protoplasts plated out directly onto the surface and from those embedded in the agarose. In most cases, embedding increased the frequency of regeneration. Protoplasts formed after incubation at 20 degrees C regenerated at a frequency of approximately 5%, as opposed to 2% for those produced at 30 degrees C. As roughly 40% of the protoplasts were anucleate, the percentage of regeneration can be estimated as about 12.5% at 20 degrees C and 5% at 30 degrees C. Protoplasting appears to be a satisfactory method of obtaining material for genetic experiments with G. abietina and A. abietis when other methods are not directly applicable. PMID- 11426866 TI - Multifunctional cytokinesis genes in Schizosaccharomyces pombe. AB - The proper division of cells is essential for the production of viable daughter cells. In plants and fungi, the dividing cell produces a cross-wall or septum that bisects the cytoplasm. For separation of the daughter cells, the septum has to be cleaved. To study the regulation of this process, we isolated mutants defective in septum cleavage. The mutants showed highly pleiotropic phenotypes and defined 17 novel genes. The deduced amino acid sequences of the products of the cloned genes exhibited homologies to various transcription regulators of other organisms. The homologies and the pleiotropic effects of the mutations on sexual development, stress response, mitotic stability, septum initiation and septum placement indicated that these genes affect cell separation indirectly, through multifunctional regulatory modules. PMID- 11426867 TI - Cytoskeleton in regenerating protoplasts and restoration of cell polarity in the yeast Saccharomyces cerevisiae. AB - The actin cables and microtubules were disrupted during protoplasting in Saccharomyces cerevisiae cells. In the process of protoplast regeneration, the cytoplasmic microtubules were the first to be restored; the actin patches remained regularly distributed under the surface of the growing protoplast. After the cell wall had been regenerated in a gelatine medium, the actin patches aggregated into clusters, which marked the site of bud development. An incomplete cell wall was the apparent cause for uncoupling between karyokinesis and cytokinesis. The presence of several nuclei in the cell gave rise to several buds emerging simultaneously and was probably related to their random positions. In haploids, however, the axial type of budding was also obvious in the reverting protoplasts. These observations suggest that actin is a structure involved in the restoration of polar growth during protoplast regeneration, and that the cell wall plays an important role in this process: in its absence, actin fails to polarise. PMID- 11426868 TI - Genetic analysis in the asexual fungus Aspergillus niger. AB - The genetics of A. niger has been developed since 1980. An overview is presented of the advances in developing methods and collecting data. Important tools have been a) the application of essentially different methods to isolate mutants, b) the adaptation to A. niger ofA. nidulans methodology for analysis of the parasexual cycle, c) the choice of marker genes, and in some cases the artificial introduction of such genes, to select homozygous segregants arising from mitotic recombination. With the use of parasexual recombination, a genetic linkage map of A. niger has been established. In total, 110 nuclear and 1 cytoplasmic (mitochondrial) markers are available. The application of A. niger genetics in applied research is illustrated by examples. PMID- 11426869 TI - Evaluation of genetic diversity of poplar genotypes by RAPD and AP-PCR analysis. AB - RAPD (randomly amplified polymorphic DNA) and AP-PCR (arbitrarily primed PCR) were utilized to establish the genetic diversity of 19 Populus genotypes. A set of 40 primers of random sequence was tested, of which 35 exhibited polymorphism. Eighteen primers generated 162 easily detectable bands between 250 and 2,500 base pairs in size, sufficient to distinguish between the genotypes. Similarity measures, cluster and multidimensional scaling analysis were performed to evaluate the RAPD and AP-PCR data. Our study demonstrated that in most instances similarity in the RAPD and AP-PCR banding patterns reflected the relationship due to origin. Nineteen primers gave a species or hybrid-specific pattern. One primer generated a specific pattern in P. euramericana. Ten primers produced specific fragments in VIF (P. alba), 4 primers in KOR (P. pyramidialis x P. berolinensis) and 4 primers in UNA and RAS (P. trichocarpa x P. deltoides). The results of this study demonstrated that RAPD or AP-PCR can be used to distinguish between poplar genotypes. PMID- 11426870 TI - Double-stranded rna elements and virus-like particles in Aspergilli. AB - Mycoviruses with double-stranded RNA (dsRNA) genomes are frequently encountered in Aspergillus isolates. A detailed study of such dsRNA elements in black Aspergillus isolates collected worldwide was carried out, and the data were analysed. The results indicate that about 10% of black Aspergilli are infected. However, the geographic distribution of infected isolates exhibits large variations; 3-13% of the isolates collected from different continents were found to carry dsRNA elements. Hybridization experiments indicated that electrophoretic banding patterns are not reliable tools for estimating the diversity of these mycovirus genomes. Among strains representing other Aspergillus sections, dsRNA segments indicative of mycovirus infection were observed for the first time in 4 species (A. leporis, A. petrakii, A. fumigatus and A. primulinus). The latter species is able to reproduce sexually. This is the second report on the detection of naturally-occurring dsRNAs in sexually reproducing Aspergillus species. The presence of virus-like particles in these and other Aspergilli was also examined by electron microscopy. Most infected Aspergillus isolates examined were found to carry virus-like particles in the size range 36-40 nm. PMID- 11426871 TI - Variability of isozyme and rapd markers among isolates of Mucor genevensis. AB - Mucor genevensis is a dimorphic and homothallic fungal species (Zygomycetes). Ten M. genevensis strains, each strain of the recently described new homothallic species (M. meguroense and M. hachijyoensis) and strains of M. hiemalis and M. piriformis (as outgroups for numerical analysis) were investigated. Five different enzyme systems (CAT, GDH, G6D, MDH and SOD) and five 10-bp random primers were used in isoenzyme and random amplified polymorphic DNA analyses, respectively. The data from these studies were subjected to numerical analyses. Substantial intraspecific variability was detected in M. genevensis with both of the methods applied. Though both the M. meguroense strain and the M. hachijyoensis strain revealed characteristic differences, they grouped closer to the homothallic M. genevensis than to the heterothallic M. piriformis and M. hiemalis strains. PMID- 11426872 TI - Collagen-chitosan polymeric scaffolds for the in vitro culture of human epidermoid carcinoma cells. AB - A biodegradable polymer scaffold was developed using collagen and chitosan, in the form of interpenetrating polymeric network (IPN), for in vitro culture of human epidermoid carcinoma cells (HEp-2, Cincinnati). Glutaraldehyde was used as cross-linking agent for the development of scaffold. Various types of scaffolds were prepared using different proportionate mixtures of collagen and chitosan solutions in the ratio of 3:7, 4:6, 5:5, 6:4 and 7:3 (collagen:chitosan). These scaffolds were fully characterized by Fourier transform infrared spectroscopy (FT IR), differential scanning calorimetry (DSC) and Thermogravimetric analysis (TGA). Equilibrium swelling studies were carried out in phosphate buffer of physiological pH (7.4) to study its swelling characteristics at slightly alkaline pH. The scaffold that showed optimum swelling property was selected as the best scaffold for performing in vitro culture studies. In vitro culture studies were carried out using HEp-2 cells, over the selected scaffold and its growth morphology was determined through optical photographs taken at different magnifications at various days of culture. The results of the above studies suggest that the scaffolds prepared from collagen and chitosan can be utilized as a substrate to culture HEp-2 cells and can also be used as an in vitro model to test anticancerous drugs. PMID- 11426873 TI - Compressive properties of cancellous bone defects in a rabbit model treated with particles of natural bone mineral and synthetic hydroxyapatite. AB - A rabbit model was developed to evaluate the compressive mechanical properties of cancellous bone defects treated with particles of selected bone graft substitute materials. A novel feature of the model was the precise retrieval of the site of implantation. A notable finding was a 9-fold increase in the modulus of elasticity of the defect implanted with a synthetic hydroxyapatite material after 26 weeks when compared to the modulus of the trabecular bone normally at the site. The compressive modulus of lesions treated with particles of a natural bovine bone mineral (anorganic bovine bone) was closer to the normal modulus of the cancellous bone at the site. While the compressive strength of the anorganic bone particles was less than that of normal bone, the site implanted with the bone mineral particles achieved compressive strength greater than normal after 6 weeks. Moreover, the anorganic bone particles accelerated the increase in strength of the lesion, at 6 weeks exceeding the strength achieved by the untreated defect after 26 weeks. The potential problem associated with the disparity in the compressive modulus between sites implanted with the synthetic HA particles and surrounding bone is discussed. PMID- 11426874 TI - Multilayer capsules: a promising microencapsulation system for transplantation of pancreatic islets. AB - In 1980, Lim and Sun introduced a microcapsule coated with an alginate/polylysine complex for encapsulation of pancreatic islets. Characteristic to this type of capsule is, that it consists of a plain membrane which is formed during a single procedural step. With such a simple process it is difficult to obtain instantly a membrane optimized with respect to all the properties requested for islet transplantation. To overcome these difficulties, it is recommended to build up the membrane in several consecutive steps, each optimized for a certain property. In this study, we have analysed such a multilayer microcapsule for the encapsulation of pancreatic islets. Therefore, empty and islet containing alginate beads were coated with alternating layers of polyethyleneimine, polyacrylacid or carboxymethylcellulose and alginate. By scanning electron microscopy the thickness of the covering multilayer-membrane was estimated to be less than 800 nm by comparison with an apparatus scale. Ellipsometric measurements showed that the membrane thickness is in the range of 145 nm. Neither the encapsulation procedure, nor the membrane-forming step did impede the stimulatory response of the islets. The encapsulation even lead to a significantly better stimulatory response of the encapsulated islets during week three and five of cell culture. Furthermore, the multilayer-membrane did not deteriorate the biocompatibility of the transplanted microcapsules, allowing an easy tuning of the molecular cut-off and the mechanical stability depending on the polycation-polyanion combination used. The multilayer membrane capsule has obvious advantages compared to a one-step encapsulation procedure. These beads guarantee a high biocompatibility, a precisely adjusted cut-off, an optimal insulin-response and high mechanical stability although the membrane is only 145 nm thick. PMID- 11426875 TI - Hydrophilic hybrid IPNs of segmented polyurethanes and copolymers of vinylpyrrolidone for applications in medicine. AB - The preparation and biocompatibility properties of thermoplastic apparent interpenetrating polymer networks (T-IPNs) of a segmented polyurethaneurea, Biospan (BS), and vinylpyrrolidone-dimethylacrylamide (VP-DMAm) copolymers, are described. The biological interaction between the obtained materials and blood was studied by in vitro methods. The addition of the VP-DMAm copolymers to form T IPNs with BS substantially increased the equilibrium water uptake and water diffusion coefficients. Investigation of the proteins adsorption, platelet adhesion, thrombus formation and factor XII activation is presented. Investigations of the proteins adsorption of the BS/VP-DMAm T-IPNs surfaces show that the segmented polyurethane (BS) containing VP-DMAm copolymers with higher VP content adsorb more albumin than fibrinogen and gamma-globulin. The platelets adhesion, thrombus formation and factor XII activation are effectively suppressed with respect to the segmented polyurethane when VP-DMAm copolymers with high VP contents are incorporated into BS as T-IPNs. PMID- 11426876 TI - Interactions between human whole blood and modified TiO2-surfaces: influence of surface topography and oxide thickness on leukocyte adhesion and activation. AB - An in vitro model (Nygren et al., J Lab Clin Med 129 (1997) 35-46) was used to investigate interactions between leukocytes and four modified TiO2-surfaces. Surface topography was measured using scanning electron microscopy and optical profilometry while Auger electron spectroscopy was used to determine surface composition and oxide thickness. The surfaces were either smooth or rough with either thin or thick oxides. All surfaces consisted of TiO2 covered by a carbonaceous layer. The surfaces were incubated with capillary blood for time periods of between 8 min and 32 h. Immunofluorescence techniques together with computer aided image analysis and chemiluminescence technique were used to detect cell adhesion, expression of adhesion receptors and the zymosan-stimulated respiratory burst response. Leukocyte adhesion to the surfaces increased during the first hours of blood-material contact and then decreased. Polymorphonuclear granulocytes were the dominating leukocytes on all surfaces followed by monocytes. Cells adhering to rough surfaces had higher normalized expression of adhesive receptors than cells on smooth surfaces. Maximum respiratory burst response occurred earlier on the smooth than on the rough surfaces. In conclusion, topography had a greater impact than oxide thickness on most cellular reactions investigated, but the latter often had a dampening effect on the responses. PMID- 11426877 TI - Comparison of five procedures for the purification of insoluble elastin. AB - Elastin is an insoluble, highly cross-linked protein, providing elasticity to organs like lung. aorta, and ligaments. Despite its remarkable mechanical properties. elastin has found little use as a biomaterial. Purification of intact elastin from elastic fibres presents a major challenge, among others for the intimate interwoveness of elastin and microfibrils. Insoluble elastin preparations tend to calcify, which may be due to calcium-binding microfibrillar (e.g. fibrillin). In this study, elastin was purified from horse ligamentum nuchae using five different procedures. One procedure is based on treatment with 0.1 M NaOH, another on autoclaving and treatment with cyanogen bromide. Three other procedures are based on combinations of extraction steps and enzyme digestions. Purity of preparations was assessed by sodium dodecyl sulphate polyacrylamide gel electrophoresis, amino acid analysis, bright field immunofluorescence and transmission electron microscopy. The procedure involving extractions/enzymes combined with an early application of 2-mercaptoethanol and cyanogen bromide gives a highly pure elastin preparation. Electron microscopic analysis showed that this preparation is devoid of microfibrillar components. This procedure is therefore the method of choice for preparation of insoluble elastin as a biomaterial for tissue engineering. PMID- 11426878 TI - Apatite formed on the surface of plasma-sprayed wollastonite coating immersed in simulated body fluid. AB - Wollastonite coatings on titanium alloys substrates were prepared by plasma spraying and incubated in simulated body fluids for different periods to investigate the nucleation and growth of apatite on their surface. Surface structural changes of the specimens were analyzed by XRD and IR technologies. SEM and EDS were used to observe surface morphologies and determine the composition of wollastonite coatings before and after immersion in simulated body fluid. The changes in the concentrations of calcium, silicon and phosphorus in the simulated body fluids due to the immersion of the specimens were measured by inductively coupled plasma atomic emission spectroscopy. The results obtained showed that hydroxycarbonate apatite can be formed on the surface of the coating soaked in SBF for 1 day. With longer immersion periods, the coating surface was covered by hydroxycarbonate apatite, which indicated that the wollastonite coating possesses good bioactivity. PMID- 11426879 TI - Role of acid attack in the in vitro bioactivity of a glass-ceramic of the 3CaO.P2O5-CaO.SiO2-CaO.MgO.2SiO2 system. AB - A non-bioactive glass-ceramic (GC13) that contains hydroxyapatite (Ca5(PO4)3OH), diopside (CaMg(SiO3)2) and althausite (Mg2 PO4OH) as crystalline phases has been obtained by thermal treatment of a parent bioactive glass (G13) of nominal composition (wt%) 40.0 CaO-34.5 SiO2-16.5 P2O5-8.5 MgO-0.5CaF2. To induce bioactivity, GC13 was chemically treated with 1 M HCl for different periods of time. After chemical etching the in vitro studies showed formation of an apatite like surface layer. In this article the influence of etching time both on the surface composition of the glass-ceramic and on the growth rate of the apatite layer is studied. It is concluded that the presence of hydroxyapatite in the glass-ceramic, associated to microstructural fluctuations, can favour apatite deposition in vitro. PMID- 11426880 TI - In vitro interaction of bacteria with polypropylene/ePTFE prostheses. AB - The infection of an implanted prosthetic material can have serious consequences on the tissue integration of the implant and the scarring process in the host, and may even necessitate replacement of the prosthesis. This study was designed to explore the in vitro effects of Staphylococcus aureus (Sa) and Staphylococcus epidermidis (Se) on polypropylene (PL) and expanded polytetrafluoroethylene (ePTFE) prostheses. Fragments of PL and ePTFE were placed in a medium previously inoculated with Sa, Se or Sa + Se (1 x 10(8) cfu Sa or/and Se). Bacterial effects on the biomaterials were evaluated for 30 days through scanning electron microscopy (SEM). In order to confirm the presence of bacteria on the prostheses, specimens were Gram stained and challenged with an antibody against protein-A (a specific Sa bacterial wall component). In both methods, specimens were examined by light microscopy. The presence of bacteria as microcolonies or biofilms was detected at PL filament cross-over regions after 30 days. Bacterial colonization of the ePTFE fragments was observed in internodal areas, which led to the deformation of prosthetic filaments. The present findings indicate that Sa and Se colonize the cross-over regions of the PL filaments, whereas in ePTFE prostheses, it is the internodal areas which are mostly affected. The latter areas are of difficult access to defence agents. PMID- 11426881 TI - Inhibition of human gelatinases by metals released from dental amalgam. AB - The interaction between metal ions and the oral environment is a major subject matter in dental research. Matrix metalloproteinases (MMPs) have been implicated in pathologic oral processes such as periodontal tissue destruction, root caries, tumor invasion and temporomandibular joint disorders. The aim of this study was to test the effect of metal ions released from dental amalgam on the major gingival gelatinolytic MMPs. Gingival human explants were cultured overnight in DMEM and the activity of secreted enzymes was analyzed by gelatin zymography in buffers conditioned with dispersed phase or concentional phase dental amalgams. The major enzymes present in conditioned media were characterized as MMP-2 and MMP-9 by immunoprecipitation. The proteolytic activities of MMP-2 and MMP-9 were strongly inhibited by dispersed phase amalgams conditioned buffers. Inhibition of MMP-2 and MMP-9 activities was partly prevented by the addition of 1,10 phenanthroline, a divalent metal chelator, to the amalgam conditioned buffers. Dental amalgam conditioned buffer also inhibited the degradation of denatured type I collagen by purified MMP-2 on liquid phase assays. These findings suggest that the activity of oral tissue MMPs may be modulated by metal ions released from dental amalgam. PMID- 11426882 TI - Adsorption of xenobiotics to plastic tubing incorporated into dynamic in vitro systems used in pharmacological research--limits and progress. AB - Commonly used materials incorporated into dynamic culture systems typically show the feature of adsorption of lipophilic xenobiotics. Yet, this phenomenon is strongly limiting the use of dynamic culture models and ex vivo organ perfusions in pharmacological and toxicological research. The aim of the study was to characterize different materials with respect to their capacity for drug adsorption and to find methods or materials to reduce the loss of substrate by adsorption in order to improve the use of dynamic in vitro systems. The adsorption of different xenobiotics (lidocaine, midazolam, lormetazepam, phenobarbital, testosterone, ethoxyresoroufine) to tubes used in dynamic in vitro systems (polyvinyl-chloride, silicone) were investigated and compared to a new material (silicone-caoutchouc-mixture). In addition, the role of protein deposition onto the tubing was studied and it was investigated whether it was possible to reach saturation of the inner tube surface by pre-loading it with the test compound. We found that silicone tubes provided the highest comfort with respect to handling and reusability, but they also demonstrated the highest capacity for substrate adsorption. Polyvinyl-chloride was the second best in handling but also demonstrated a high complexity in its adsorption behavior. The silicone-caoutchouc-mixture reached acceptable experimental results with respect to its handling and demonstrated a very low capacity for substrate adsorption. PMID- 11426883 TI - The effect of particle size on the gelation of tissue conditioners. AB - Tissue conditioner materials comprise a methacrylate polymer powder, usually poly(ethyl methacrylate) (PEM), and a plasticiser that also contains ethanol. The aim of this study was to investigate the effect of ball milling the PEM on gelation time. PEM was used as received and after ball milling for 2, 4, 8 and 16 h. Particle size was measured in each case and gelation time at 37 degrees C when mixed with butyl phthalyl butyl glycollate (BPBG). Gelation time reduced with increasing milling time of the powder, however it was found that increase in the fines initially had a more significant affect than overall reduction in average particle size. The gelation time of unmilled and 16 h milled PEM were also measured when mixed with BPBG and various levels of ethanol. Unmilled PEM with 4% ethanol had a gelation time of 13.5 +/- 2.0 min compared to 5.5 +/- 1.0 min for the 16h milled PEM which was similar to that for the commercial material, Viscogel (6.0 +/- 1.0 min). Ball milling the PEM reduces the level of ethanol required to produce a clinically acceptable gelation time. PMID- 11426884 TI - Influence of surface modifications to titanium on antibacterial activity in vitro. AB - The antibacterial effect of surface modifications to titanium on Porphyromonas gingivalis ATCC 33277 and Actinobacillus actinomycetemcomitans ATCC 43718 was evaluated. Surface modifications were performed with dry processes including ion implantation (Ca+, N+, F+), oxidation (anode oxidation, titania spraying), ion plating (TiN, alumina), and ion beam mixing (Ag, Sn, Zn, Pt) with Ar+ on polished pure titanium plates. F+-implanted specimens significantly inhibited the growth of both P. gingivalis and A. actinomycetemcomitans than the polished titanium. The other surface-modified specimens did not exhibit effective antibacterial activity against both bacteria. No release of the fluorine ion was detected from F-implanted specimens under dissolution testing. This result and the characterization of the F+-implanted surfaces suggested that the possible antibacterial mechanism of the F+-implanted specimen was caused by the formation of a metal fluoride complex on the surfaces. In addition, F+-implanted surfaces did not inhibit the proliferation of fibroblast L929-cells. These findings indicate that surface modification by means of a dry process is useful in providing antibacterial activity of oral bacteria to titanium implants exposed to the oral cavity. PMID- 11426885 TI - Indomethacin-loaded methoxy poly(ethylene glycol)/ poly(epsilon-caprolactone) diblock copolymeric nanosphere: pharmacokinetic characteristics of indomethacin in the normal Sprague-Dawley rats. AB - We prepared the drug-loaded polymeric nanospheres composed of the methoxy poly(ethylene glycol) (MePEG) and poly(epsilon-caprolactone) (PCL) that showed a narrow size distribution and average diameter of less than 200 nm. We could obtain the nanosphere having a relatively high drug-loading efficiency of about 42% when the feed weight ratio of indomethacin (IMC) to polymer was 1:1. To investigate the IMC pharmacokinetics in the IMC-loaded MePEG/PCL nanosphere (DMEP70) using the rats as animal model, we analyzed the IMC concentration in plasma with HPLC after i.v. bolus administered at a dose of 10 mg/kg in free IMC (control) and IMC-loaded MePEG/PCL nanosphere (DMEP70) groups via tail vein. Pharmacokinetics parameters (mean +/- s.d.) such as the mean residence time (MRT, h), the steady-state volume of distribution (Vdss, l), the terminal half-time (t 1/2, h) and the plasma clearance (CL, l/h) of IMC in each groups (control vs. DMEP70) were determined; MRT (16.97 +/- 4.83 vs. 28.69 +/- 11.28, p < 0.01); Vdss (14.26 +/- 4.86 vs. 20.37 +/- 12.04, p < 0.05); t 1/2 (15.12 +/- 4.77 vs. 23.1 +/ 8.24, p < 0.01); CL (0.84 +/- 0.27 vs. 0.71 +/- 0.41). From these results, we could concluded that MEP70 has a significant potential for sustained release and the enhancement of circulation time of loaded drug by prolonging terminal half life, increasing MRT and Vdss of IMC. Therefore, The MePEG/PCL block copolymeric nanosphere system is being considered as promising biodegradable and biocompatible drug carrier vehicles for parentral use and may be useful as sustained release injectable delivery systems for hydrophobic drugs. PMID- 11426886 TI - Biocompatibility testing of novel starch-based materials with potential application in orthopaedic surgery: a preliminary study. AB - This paper describes an extensive biocompatibility evaluation of biodegradable starch-based materials aimed at orthopaedic applications as temporary bone replacement/fixation implants. For that purpose, a polymer (starch/ethylene vinyl alcohol blend, SEVA-C) and a composite of SEVA-C reinforced with hydroxyapatite (HA) particles, were evaluated in both in vitro and in vivo assays. For the in vitro analysis cell culture methods were used. The in vivo tissue reactions were evaluated in an intramuscular and intracortical bone implantation model on goats, using light and scanning electron microscopy. A computerized image analysis system was used to obtain histomorphometric data regarding bone contact and remodelling after 6 and 12 weeks of implantation. In both in vitro and in vivo models, the SEVA-C-based materials did not induce adverse reactions, which in addition to their bone-matching mechanical properties makes them promising materials for bone replacement fixation. PMID- 11426887 TI - Down's syndrome screening: where to now? PMID- 11426888 TI - Evaluation of a clinical test. I: assessment of reliability. PMID- 11426889 TI - Evaluation of a clinical test. II: Assessment of validity. PMID- 11426890 TI - The reliability and validity of three dimensional ultrasound volumetric measurements using an in vitro balloon and in vivo uterine model. AB - OBJECTIVE: To evaluate the reliability and validity of two and three dimensional ultrasound volumetric measurements using balloon and uterine models. DESIGN: Prospetive observational study. SETTING: Obstetric ultrasound department at a university teaching hospital. METHOD: Two and three dimensional ultrasound volumetric measurements (with 5, 10 and 15 ultrasonic slices) were performed on 30 different sets of ultrasound images obtained from 15 water filled balloons with volumes ranging from 19 to 697mL. The measurements were performed independently by two observers who were blinded to the true volumes of the balloons. For the uterine model, only three dimensional ultrasonic volume measurements were performed independently on 16 uteri by two observers who were again unaware of the definitive uterine volumes. OUTCOME MEASURE: For the assessment of intra-and inter-rater reliability, the intraclass correlation coefficient was used. The index of concordance between the ultrasonic volumes and those obtained by the reference standard (validity) was assessed with the conventional Pearson's correlation coefficient, limits of agreement method and the intra-class correlation coefficient. RESULTS: High levels of reliability and validity were obtained for both two and three dimensional ultrasound balloon volume measurements. For two dimensional ultrasonic volume measurements, the intra-class correlation coefficient ranged from 0.992 to 0.998 for reliability and validity whereas the Pearson's correlation coefficient for validity was 0.996. With three dimensional ultrasonic volume measurements, the intra-class correlation coefficient ranged from 0.991 to 0.999 for reliability and validity whereas the Pearson's correlation coefficient for validity was 0.999. Both two and three dimensional ultrasonic measurements tended to underestimate the true balloon volume with the largest observed mean difference obtained with three dimensional ultrasound measurements using five ultrasonic slices and the smallest value obtained with three dimensional ultrasound measurements employing 15 ultrasonic slices. The mean difference in volume measurement for two dimensional ultrasound was intermediate between these two values. However, two dimensional ultrasound volume measurement generated the largest range between the limits of agreement whereas the smallest range was obtained with three dimensional ultrasound using 10 ultrasonic slices. The intra-class correlation coefficient for reliability and validity with three dimensional ultrasonic uterine volume estimation ranged from 0.956 to 0.996 whereas the Pearson's correlation coefficient for validity ranged from 0.993 to 0.999). The use of three dimensional ultrasound also consistently under-estimated the actual uterine volumes. The larger the number of ultrasonic slices employed for three dimensional ultrasound, the smaller was the mean difference between the ultrasonic and true uterine volume measurements and the smaller the limits of agreement. CONCLUSIONS: The reliability and validity of balloon and uterine volume measurement by three dimensional ultrasound is high. This allows further research on three dimensional ultrasound for measuring pelvic organ volumes in the prediction of pelvic pathology. PMID- 11426891 TI - The accuracy of lamellar body count and lecithin/sphingomyelin ratio in the prediction of neonatal respiratory distress syndrome: a meta-analysis. AB - OBJECTIVE: To compare the performance of the lecithin/sphingomyelin ratio and the lamellar body count in the prediction of neonatal respiratory distress syndrome. DESIGN: Meta-analysis. SAMPLE: Six studies reporting on the performance of both the lecithin/sphingomyelin ratio and the lamellar body count published between January 1966 and August 1999. METHODS: We performed a computerised MEDLINE search to identify articles published on the subject. For the six selected studies, prevalence of respiratory distress syndrome and sensitivity and specificity of the tests in the prediction of respiratory distress syndrome were calculated, and overall performance was assessed by constructing summary receiver-operating characteristic curves. RESULTS: The constructed summary receiver-operating characteristic curves showed the lamellar body count to perform slightly better than the lecithin/sphingomyelin ratio in the prediction of respiratory distress syndrome (P= 0.13). CONCLUSIONS: Since the lamellar body count can be performed quickly and since it is less expensive than the lecithin/sphingomyelin ratio, we recommend the former as the test of first choice in the assessment of fetal lung maturity. PMID- 11426892 TI - False positive serum biochemical screening and subsequent fetal loss in women less than 35 years of age. AB - OBJECTIVE: To examine the fetal loss rate in women younger than 35 years of age following a false positive serum biochemical screening. DESIGN: Retrospective analysis of case records between 1991 and 1998. SETTING: Fetal medicine unit of a large teaching hospital. POPULATION: Four hundred and fifty-six women with singleton pregnancies and false positive serum biochemical screening for Down's Syndrome (study group). Nine hundred and twelve matched controls with true negative serum biochemical screening (control group). METHODS: Women of both groups had a second trimester serum screening for Down's Syndrome using alpha fetoprotein, human chorionic gonadotrophin (hCG) and unconjugated oestriol (uE3); and they also underwent genetic amniocentesis. RESULTS: The overall fetal loss rate in the study group was 5.3% (24/456), compared with 1.65% (15/912) in the control group RR 3.2, 95% CI 1.7-5.99; P < 0.001). The majority of fetal losses in the study group occurred after 28 weeks, while in the controls this happened between 24 and 28 weeks of gestation. CONCLUSIONS: A false positive serum biochemical screening in women under 35 years of age is associated with a threefold increased risk of subsequent fetal loss. However, most of fetal losses in this group occurred after 28 weeks, indicating that intensive antepartum fetal surveillance could improve the perinatal outcome. PMID- 11426893 TI - Fever in labour and neonatal encephalopathy: a prospective cohort study. AB - OBJECTIVE: To determine whether the reported association of maternal fever with neonatal encephalopathy is independent of other associated intrapartum risk factors. DESIGN: Prospective cohort study. SETTING: Dublin teaching hospital delivery ward. POPULATION: 4,915 low risk women in labour at 36-41 weeks of gestation. METHODS: Using logistic regression with odds ratios and 95% confidence intervals, the incidence of neonatal encephalopathy and other neonatal outcomes of women who had an intrapartum fever >37.5 degrees C was compared with those who did not. RESULTS: The cohort comprised 33% of all deliveries during the study period. Neonatal encephalopathy was diagnosed in 3.25/1,000 births. The incidence of intrapartum fever was 6.8%. Maternal fever was strongly associated with neonatal encephalopathy (crude OR 10.8, 95% CI 4.0-29.3). Univariate analysis showed maternal fever was associated with epidural analgesia, nulliparity, induction, longer labour, oxytocin administration, greater fetal birthweight and gestational age and instrumental vaginal delivery, but not with prolonged (>24hours) prelabour rupture of the membranes. The association of fever with neonatal encephalopathy persisted having adjusting for these covariates (adjusted OR 4.72, 95% CI 1.28-17.4). CONCLUSIONS: The relationship between maternal intrapartum fever and neonatal encephalopathy is independent of other known intrapartum risk factors. This provides further evidence for the role of inflammatory processes in the aetiology of neonatal neurological morbidity. PMID- 11426894 TI - Are there common triggers of preterm deliveries? AB - OBJECTIVE: To assess the effect(s) of transient events which are perceived as stressful on the inseption of preterm delivery. DESIGN: A case-control study, with immature infants as cases and borderline term babies as controls. SETTING: A teaching maternity hospital in Athens. POPULATION: All infants born at less than 37 weeks of gestation, during a twelve-month period. METHODS: Information was collected about maternal socio-demographic and lifestyle characteristics, clinical variables and stressful events occurring within two weeks prior to delivery. MAIN OUTCOME MEASURES: Factors affecting the risk of preterm delivery. RESULTS: Extreme prematurity (<33 weeks) is more common among younger (<25 years of age) and older (>29 years of age) women and is positively associated with parity, body mass index and smoking, whereas it is inversely associated with educational level, regular physical exercise and serious nausea/vomiting. After controlling for these factors, however, only coitus during the last weeks of pregnancy had a significant triggering effect on prematurity (P = 0.004, odds ratio 3.21, 95% CI 1.45 to 7.09 for very immature babies, and P = 0.04, OR = 2.20, 95% CI 1.03 to 4.70 for immature babies). On the contrary, several events perceived as stressful, such as illness of relatives or friends, husband's departure, loss of employment, were unrelated to the onset of premature labour. CONCLUSIONS: Coitus during the last few weeks of pregnancy appears to increase the risk of preterm delivery, while a possible detrimental effect of physical exertion seems more limited. Stressful events should not receive undue attention as possible causes of preterm delivery. PMID- 11426895 TI - Beta-3 versus beta-2 adrenergic agonists and preterm labour: in vitro uterine relaxation effects. AB - OBJECTIVE: 1. To investigate the effects of the selective beta-3 adrenoreceptor agonist, BRL 37344, on human pregnant myometrial contractility in vitro. 2. to compare these effects with those of the beta-2 adrenoreceptor agonist, ritodrine. METHODS: Isometric tension recording was performed under physiological conditions in isolated myometrial strips from biopsies obtained at elective caesarean section. Following pre-incubation with oxytocin (10(-9) M), the effects of cumulative additions of BRL 37344 or ritodrine (10(-8)-10(-3.5) M) on myometrial contractility were investigated. Results were expressed as -log EC50 (pD2) and mean maximal inhibition achieved for both drug compounds. RESULTS: BRL 37344 exerted a concentration dependant relaxant effect on myometrial contractions in all strips exposed [pD2, 7.26 (0.48) (SEM); mean maximal inhibition 61.98 (4.89%); n = 6]. Similarly, ritodrine exerted a concentration dependant inhibition of myometrial contractility in all strips exposed [pD2 = 7.40 (0.28); mean maximal inhibition 59.49 (3.97%); n = 6]. There was no significant difference between calculated pD2 values (P = 0.65) or mean maximal inhibition achieved (P = 0.79). CONCLUSIONS: The beta-3 adrenoreceptor agonist BRL 37344 induced relaxation of human myometrial contractions with similar potency to that of the most commonly used tocolytic agent ritodrine. This raises the possibility that the novel beta-3 adrenoreceptor agonists may have potential as therapeutic agents for human preterm labour. In view of their reported reduced cardiovascular side effects their potential clinical use requires further evaluation. PMID- 11426896 TI - Microvascular vasodilator response to acetylcholine is increased in women with pre-eclampsia. AB - OBJECTIVES: To evaluate in vivo microvascular responses to incremental doses of the endothelial-dependent vasodilator, acetylcholine, and the endothelial independent vasodilator, sodium nitroprusside, in women with pre-eclampsia and gestation-matched normotensive pregnant controls. DESIGN: Prospective clinical study. SETTING: Southern Derbyshire Acute Hospitals Trust, and University of Nottingham Division of Vascular Medicine. POPULATION: Thirteen women with pre eclampsia and 16 normotensive pregnant controls. METHODS: Cutaneous microvascular vasodilator responses to iontophoretic administration of incremental doses of acetylcholine and sodium nitroprusside (25-100 microC) were evaluated under temperature-controlled conditions using laser Doppler fluximetry. RESULTS: Resting skin temperature and blood flow were similar among 13 women with pre eclampsia [mean blood pressure 151/93 mmHg (4/3); mean gestation 35.6 weeks (1.0); and mean proteinuria 1.1 g/24h (0.2)] and 16 normotensive pregnant controls [mean blood pressure 111/63 mmHg (2/2); mean gestation 34.3 weeks (0.9)]. Peak vasodilator responses to acetylcholine were increased in women with pre-eclampsia (median flux ratio 15.1 [IQR 12.3-17.6] vs 11.7 [IQR 8.4-12.6], P < 0.05), whereas sodium nitroprusside responses were not different between the two groups: 11.4 [IQR 8.6-13.4] vs 9.5 [IQR 8.0-12.3]. CONCLUSION: In vivo microvascular vasodilator responses to acetylcholine are increased in women with pre-eclampsia, while endothelial-independent vasodilation is unchanged. Although the mechanism of acetylcholine induced vasodilation in small vessels is unclear, this study confirms previous animal data and provides in vivo evidence of altered microvascular endothelial cell function in pre-eclampsia. PMID- 11426897 TI - Leucocyte intracellular pH and Na+/H+ exchanger isoform-1 activity in postpartum women with pre-eclampsia. AB - OBJECTIVE: To investigate leucocyte Na+/H+ exchanger isoform 1 activity in postpartum pre-eclamptics. DESIGN: Exchanger isoform-1 activity and intracellular resting pH were established in leucocytes isolated from two study groups. SAMPLE: Leucocytes isolated from 10 women who had had pre-eclamptic pregnancies more than five months postpartum, and from 10 age-matched normotensive women who were more than five months postpartum. SETTING: Hypertension Clinic, Antenatal Assessment Area, Leicester Royal Infirmary. METHODS: A well validated technique involving flurometry using a pH sensitive dye (BCECF-AM) was performed to determine exchanger isoform-1 activity and intracellular pH. Determination of exchanger isoform-1 protein abundance was performed by western blotting. Exchanger isoform 3 protein abundance was examined to rule out the possibility of activity due to this particular isoform. RESULTS: Intracellular pH was significantly lower in the postpartum pre-eclamptic group (7.11 +/- 0.02), compared with the postpartum normotensive controls (7.33 +/- 0.04; P < 0.001). Exchanger isoform-1 efflux rate (in mmol/L/minute) was significantly higher in the postpartum pre-eclamptic group (35.91 +/- 3.1), compared with the postpartum normotensives (23.94 +/- 2.0; P = 0.005). Exchanger isoform-1 protein density was established to be similar among the two subject groups. No exchanger isoform-3 protein was identified by western blotting. CONCLUSION: Our results suggest that elevated exchanger isoform-1 activity is an important finding in women who have suffered from pre-eclampsia. This increased activity is not due to an increase in exchanger isoform-1 protein abundance or the presence of exchanger isoform-3. PMID- 11426898 TI - Premenopausal women affected by sexual arousal disorder treated with sildenafil: a double-blind, cross-over, placebo-controlled study. AB - OBJECTIVE: To verify whether sildenafil is effective in young premenopausal women affected by arousal disorder. DESIGN: A double-blind cross-over study. SETTING: Centre for Sexological Research, Department of Microbiological and Gynaecological Science, University of Catania, Italy. SAMPLE: Fifty-three volunteer women aged 22-28 years affected by arousal disorders. METHODS: The study consisted of three 4-week periods: sildenafil, washout, placebo, by six possible sequences. sildenafil was used at 25 mg or 50 mg. MAIN OUTCOME MEASURES: Efficacy was assessed at baseline and once monthly by the Personal Experiences Questionnaire based on the 5-point Likert scale. The questionnaire quantified subjective arousal (primary endpoint), and orgasm, enjoyment, sexual frequency, and the number of sexual fantasies. RESULTS: Fifty-one women completed the study. Mean (SD) usage of sildenafil 25mg and 50mg was, respectively, 2.8 (0.8) and 2.7 (1.3) times weekly, while mean usage of placebo was 2.8 (1.6) times weekly. During both sildenafil dosages, arousal and orgasm improved with respect to placebo (P < 0.001). Therapeutically significant differences were not noted during the treatment with both 50 mg and 25 mg of sildenafil for arousal and orgasm. The frequency of sexual fantasies and of sexual intercourse, and enjoyment, improved in the women treated with sildenafil (P < 0.05). CONCLUSIONS: Our study suggests that sildenafil may improve sexual performance of women affected by sexual difficulties such as arousal disorder, and may indirectly improve other aspects of sexual life. Moreover, further studies need to define the use of PDE type 5 inhibitors in this sexual pathophysiology. PMID- 11426899 TI - Sacrohysteropexy with synthetic mesh for the management of uterovaginal prolapse. AB - OBJECTIVE: To study the ongoing results of sacrohysteropexy with Teflon mesh for treatment of uterovaginal prolapse in women who desire to preserve their uterus. DESIGN: Prospective observational study. SETTING: Tertiary referral urogynaecology unit. PARTICIPANTS: Thirteen consecutive women with uterovaginal prolapse wishing to retain their uterus operated on by one surgeon. SURGICAL METHOD: Sacrohysteropexy with Teflon mesh attached to the uterine isthmus and to the anterior longitudinal ligament of the first or second sacral vertebra in a tension-free fashion. MAIN OUTCOME MEASURES: Subjective and objective cure of uterine prolapse and operative and post-operative complications. RESULTS: The mean age of the women was 38 years (range 27-60). Eight women were multiparous. Twelve women had second degree uterine prolapse and one woman had third degree of uterine prolapse. Mesh was extended to correct a cystocele in one woman and a rectocele in three women. In four women colposuspension was performed at the same time. There were no intra- and post-operative complications. The mean follow up time was 16 months (range 4-49). At follow up only one woman had a first degree uterine prolapse. A total of seven women (53.8%) reported constipation which had been experienced pre-operatively by four women (30.8%). CONCLUSIONS: We consider the sacrohysteropexy with Teflon mesh a safe, effective and durable surgical procedure for the management of uterovaginal prolapse in young women and those who desire to retain their uterus. PMID- 11426900 TI - Factors associated with nucleic acids related to human immunodeficiency virus type 1 in cervico-vaginal secretions. AB - OBJECTIVE: To assess HIV-related nucleic acids in cervico-vaginal secretions and the factors associated with them. DESIGN: Observational study. SETTING: Department of Obstetrics and Gynaecology, University of Pavia, Italy. POPULATION: HIV-positive patients attending a cytology service. METHODS: Paired blood and cervico-vaginal lavage samples were obtained from 122 known HIV-seropositive patients during periodic visits for cytologic screening for lower genital tract neoplasia. Vaginal specimens for the diagnosis of bacterial vaginosis, trichomonas vaginalis and candida infection were also obtained. HIV-1-RNA in plasma, proviral HIV-1-DNA, cell associated and cell-free HIV-1 RNA in cervico vaginal secretions were quantitatively evaluated by competitive polymerase chain reaction (c-PCR) and reverse transcriptase PCR (cRT-PCR). MAIN OUTCOME MEASURE: Prevalences of HIV related nucleic acids in cervico-vaginal secretions and their univariate and multivariate associations with clinical variables. RESULTS: Proviral HIV-1 DNA, cell-associated and cell-free HIV-1 RNA were detected in 50% (61/122), 37.7% (46/122) and 32.8% (40/122) of the patients, respectively. In logistic regression analysis, the presence of HIV-1 RNA in blood was the factor which correlated best with the detection of HIV-1 DNA (OR = 5.48, 95% CI = 2.28 13.20), cell-associated (OR = 4.85; 95% CI = 1.89-12.45) and cell-free HIV-1 RNA (OR = 4.63, 95% CI = 1.74-12.33) in cervico-vaginal samples. However, between 20% and to 35% of patients who tested negative for blood HIV-1 RNA were positive for either HIV-1 DNA or HIV-1 RNA detection in cervico-vaginal lavages. Bacterial vaginosis was associated with an increased prevalence of cell-associated (OR = 3.58, 95% CI = 1.22-10.54) and cell-free HIV-1 RNA (OR = 2.94, 95% CI = 1.0-8.7) detection in cervico-vaginal secretions. Additional factors associated with increased prevalence of HIV-1 RNA detection were advanced stage of HIV disease and vulvovaginal candidiasis. CONCLUSIONS: Although the presence of HIV-1 RNA in blood is the factor which correlates best with the detection of HIV-related nucleic acids in cervico-vaginal secretions, the shedding of HIV in the genital tract can occur in 20-30% of non-viremic subjects. Bacterial vaginosis and candida infection could have a facilitating role in local HIV viral replication and shedding. PMID- 11426901 TI - The role of comparative genomic hybridisation in prenatal diagnosis. AB - The aims of this study were to assess the feasibility of using comparative genomic hybridisation instead of conventional cytogenetics in prenatal diagnosis and to determine the size of DNA loss that can be detected. Using comparative genomic hybridisation, six cases with standard aneuploidies were diagnosed correctly. This technique clearly identified a partial duplication of the long arm of chromosome 1 but was not capable of detecting the associated inversion. A small interstitial deletion on short arm of chromosome 10 also was detected precisely. Although the current comparative genomic hybridisation resolution is similar to the sensitivity of the highest resolution G banding, the latter is not a routine strategy in prenatal diagnosis. Comparative genomic hybridisation can allow full chromosome assessment equal to the highest resolution cytogenetic studies without the need for cell culture. PMID- 11426902 TI - Lay persons' understanding of the risk of Down's syndrome in genetic counselling. AB - Genetic counselling traditionally expresses risk in proportions (e.g. 1 in 112) rather than as rates (e.g., 8.9 per 1,000). The justification for this practice is unclear. To assess the understanding of lay persons of the risk of Down's Syndrome, whether expressed as rates or as proportions, we analysed 589 self administered questionnaires. Overall, respondents understood rates significantly better than proportions (76.2% vs 72.3% correct, respectively; P = 0.03) Evidence from two studies in disparate populations suggests that rates are better understood and thus are the preferred way to explain genetic risk to lay persons. PMID- 11426903 TI - A novel in vitro co-culture system for the study of maternal decidual endothelial cell-trophoblast interactions in human pregnancy. AB - Investigation of the pathophysiology of pre-eclampsia (characterised by insufficient invasion of the intrauterine vasculature by cytotrophoblasts) has been hampered by the absence of a suitable animal model, and ethical constraints in clinical studies. We have developed a novel in vitro human cell co-culture system allowing direct assessment of cytotrophoblast invasion of a decidual endothelial cell monolayer from the abluminal side, as occurs in vivo. This model will facilitate detection, at the cellular level, of abnormal endothelial cell trophoblast functional interactions in pre-eclampsia and other pregnancy disorders with abnormal placentation. PMID- 11426904 TI - Screening for HIV, hepatitis B and C infection in a population seeking assisted reproduction in an inner London hospital. AB - The Human Fertilisation and Embryology Authority requires all sperm donors to be screened for human immunodeficiency virus (HIV), hepatitis B and C and their semen quarantined for six months. No guidelines exist for screening prior to in vitro fertilisation or intracytoplasmic sperm injection. We prospectively analysed the prevalence of these viruses in our patients. Screening detected one case of HIV (0.13%), four of hepatitis C (0.5%) and 14 new cases of hepatitis B (1.7%). The prevalence of hepatitis B and HIV in our antenatal population at this time was 1.4% and 0.8%, respectively. Knowledge allows measures to be taken to reduce the risk of transmission to partner, fetus, new born baby, or by cross contamination during embryo cryostorage and enables couples to make an informed decision regarding proceeding with treatment. Detection of infection in one partner should no longer preclude fertility treatment. PMID- 11426905 TI - Spontaneous opening of the Filshie clip as a cause of sterilisation failure. AB - An open Filshie clip is sometimes detected during the evaluation of a case of sterilisation failure. This spontaneous opening of the clip does not need to have been caused by material failure, but may be produced by jamming of the clip what can occur by incorrect squeezing during closure. On superficial observation, the clip will appear to be closed, but it may easily open itself by its spring mechanism when the clip is moved or merely touched. Especially in the teaching situation one should be careful for this lapse. PMID- 11426906 TI - Accidental delivery through a vaginal incision (laparoelytrotomy) during caesarean section in the second stage of labour. PMID- 11426907 TI - Disseminated leiomyomatosis peritonealis in association with oestrogen secreting ovarian fibrothecoma. PMID- 11426908 TI - Vulval lichen planus: progression of pseudoepitheliomatous hyperplasia to invasive vulval carcinomas. PMID- 11426909 TI - Menorrhagia due to Bernard-Soulier syndrome and its successful treatment by thermal balloon endometrial ablation. PMID- 11426910 TI - Venous malformation of the vulva. PMID- 11426911 TI - The Antileukotriene Working Group consensus conference overview. PMID- 11426912 TI - Safety of antileukotriene agents in asthma management. AB - OBJECTIVE: This article presents information on the safety of zafirlukast, montelukast, and zileuton, three antileukotrienes (anti-LTs) approved in the United States for the prophylaxis and treatment of asthma. After reading this article, readers should have an understanding both of the general safety of anti LTs and their specific adverse effects. DATA SOURCES: Relevant and appropriate controlled clinical studies on the safety of anti-LTs in asthma were used. Only literature in the English language was reviewed. STUDY SELECTION: Material was taken from academic/scholarly journals and appropriate reviews. RESULTS: Antiasthma agents, including corticosteroids, beta2-agonists, and methylxanthines, may be categorized into two classes: those used for the long term control and prevention of persistent asthma and those used for the prompt relief of acute symptoms and exacerbations of the disease. Although most agents are safe and well tolerated when used properly, adverse effects may occur with use at higher dose levels. The anti-LTs, including zafirlukast, montelukast, and zileuton, are the first new pharmacologic class in the therapeutic armamentarium for asthma management to be approved in the United States in the past 20 years. Both zafirlukast and montelukast carry pregnancy category B classification whereas zileuton carries pregnancy category C classification. The most common adverse effects observed in clinical trials were headache, pharyngitis, abdominal pain, dyspepsia, and cough. CONCLUSIONS: The results of clinical trials and real world experience indicate that these agents are generally safe and well tolerated, with an incidence of adverse effects comparable with placebo. PMID- 11426913 TI - Adherence to asthma therapy. AB - OBJECTIVE: This article presents information on the important impact the lack of adherence to prescribed treatments has on the morbidity and mortality of asthma. After reading this article, readers should have an understanding of the significant role clinicians, as well as patients and their families, play in promoting adherence. DATA SOURCES: A detailed literature search was conducted. Relevant studies were used. Only literature in the English language was reviewed. STUDY SELECTION: Material was taken from academic/scholarly publications, appropriate reviews, and published abstracts. RESULTS: Over the last decade, many researchers have examined ways to improve adherence to asthma therapy, including improving clinician-patient communication, educating the patient, and simplifying the treatment regimen. Nonadherence is a problem in pediatric and adolescent patients, whose needs are different from those of adult patients. Because proper technique is essential with metered-dose inhalers and peak flow meters, special efforts are required by clinicians to promote adherence to these devices. CONCLUSIONS: Promoting adherence involves a good clinician-patient relationship, as well as provision of personalized, practical, and repeated education. PMID- 11426914 TI - The role of antileukotrienes in the treatment of asthma. AB - OBJECTIVE: This article reviews the literature on the role of antileukotrienes (anti-LTs), specifically montelukast, zafirlukast, and zileuton, in the treatment of asthma. DATA SOURCES: Relevant and appropriate controlled clinical studies were used. Only literature in the English language was reviewed. STUDY SELECTION: Material was taken from academic/scholarly journals, appropriate reviews, and published abstracts. RESULTS: In guidelines established by the National Asthma Education and Prevention Program and the National Heart, Lung, and Blood Institute, a stepwise approach to asthma management is recommended, with recommendations varying depending on degree of disease severity. The anti-LTs, the newest class of drugs for the treatment of asthma, play a circumscribed role in the guidelines as they were only recently available when the latest guidelines were published. Subsequently, however, extensive clinical experience with the anti-LTs has been amassed. Multiple clinical studies have demonstrated that the anti-LTs improve pulmonary function and quality of life, and reduce asthma symptoms, asthma exacerbations, and use of beta2-agonists and oral steroids. The anti-LTs may be particularly useful in asthma patients with aspirin sensitivity or concomitant allergic rhinitis, as well as in pediatric patients. These agents have additive effects with inhaled corticosteroids and may permit a reduction in inhaled corticosteroid dosages. CONCLUSIONS: The anti-LTs have several features that are likey to promote adherence to treatment and are generally well tolerated. The available clinical data suggest that anti-LTs should be considered as a therapeutic option or as additive therapy in patients with mild to severe asthma. PMID- 11426915 TI - Discovery of leukotrienes and development of antileukotriene agents. AB - OBJECTIVE: This article presents information on the origin of leukotrienes (LTs) and the development of antileukotriene (anti-LT) agents. After reading this article, readers should have an understanding of the chemical mediators involved in the pathogenesis of asthma, the structural features of LTs, and the role of anti-LTs in the management of asthma symptoms. DATA SOURCES: Studies considered relevant and appropriately controlled were used. Only literature in the English language was reviewed. STUDY SELECTION: Material was taken from academic/scholarly journals and abstracts. RESULTS: One of the important chemical mediators implicated in the pathogenesis of asthma is the slow-reacting substance of anaphylaxis, which was subsequently found to comprise LTs C4, D4, and E4. 5 lipoxygenase products from arachidonic acid metabolism, LTs are released from the lung tissue of asthmatic patients and purified human lung mast cells by antigens. The LTs directly induce contraction of bronchial smooth muscle. The use of anti LT agents, particularly the receptor antagonists zafirlukast and montelukast and the biosynthesis inhibitor zileuton, reverses the bronchoconstrictive effects of LTs and significantly improve asthma symptoms. CONCLUSIONS: Extensive in vitro and in vivo evidence supports the role of LTs in the pathogenesis of asthma. Their discovery has had a significant impact on treatment strategies, including the use of anti-LT agents, for the management of asthma. PMID- 11426916 TI - Consensus guidelines for asthma therapy. AB - OBJECTIVE: This article provides information on the consensus reached by the Antileukotriene Working Group on the role of leukotriene (LT) modifiers in the treatment of asthma. DATA SOURCES: Relevant and appropriate controlled clinical studies were used. Only literature in the English language was reviewed. STUDY SELECTION: Material was taken from academic/scholarly journals and appropriate reviews. RESULTS: Only limited use of LT-modifying agents has been recommended in recently published guidelines of the National Asthma Education and Prevention Program and the National Heart, Lung, and Blood Institute. Consequently, the Antileukotriene Working Group was convened to arrive at a consensus on the wider use of LT modifiers in the treatment of asthma. The group' s purpose was 2-fold: to review and disseminate information on the role of LT modifiers in clinical practice. As determined by the group, a thorough understanding of the patient's disease, patient education, and an effective patient-clinician relationship are key elements in overall patient management. CONCLUSIONS: Based on evidence from clinical trials and expert opinions of participants comprising the Antileukotriene Working Group, LT-modifying agents potentially may be used as first-line therapy, in combination regimens, and as an alternative to inhaled corticosteroids in the treatment of asthma. PMID- 11426917 TI - Efficacy of antileukotriene agents in asthma management. AB - OBJECTIVE: This article reviews information on the role of three antileukotrienes (anti-LTs), montelukast, zafirlukast, and zileuton, in the management of asthma. After reading this article, readers should have an understanding of the efficacy of anti-LTs in specific patient populations. DATA SOURCES: Relevant and appropriately controlled clinical studies on the efficacy of anti-LTs were used. Only literature in the English language was reviewed. STUDY SELECTION: Material was taken from peer-reviewed journals and published abstracts. RESULTS: The efficacy of anti-LTs has been established in numerous randomized, controlled, multicenter trials involving patients with mild-to-moderate disease. These agents reduce asthma symptoms, beta2-agonist use, and asthma exacerbations, in addition to improving pulmonary function. The anti-LTs are also effective when added to low-dose or high-dose corticosteroid regimens in symptomatic asthma patients. They have proven efficacy in patients with aspirin-sensitive asthma, and they appear able to reduce nasal congestion in asthma patients with concomitant upper airway symptoms. Some anti-LTs have been shown to be effective in pediatric asthma patients. CONCLUSIONS: In reviewing available clinical results as well as real-world experience in managing asthma patients, the Antileukotriene Working Group concludes that anti-LTs may be used in patients with mild persistent asthma as well as in combination with other asthma medications at all levels of disease severity for long-term maintenance of asthma control. PMID- 11426918 TI - Pesticides as a cause of occupational skin diseases in farmers. AB - Pesticides are chemical substances used in agricultural production to protect crops against pests. They help to achieve better quality and quantity of crops; however, they also are capable of causing occupational diseases in farmers. Skin is the most exposed organ while spraying the pesticide on fields. Farmers are also exposed to pesticides while mixing, loading the pesticide as well as while cleaning the equipment and disposing of empty containers. Other activities associated with exposure are sowing pesticide-preserved seeds, weeding and harvesting previously sprayed crops. During the first decades of using pesticides the main problem was the risk of acute intoxication among people occupationally exposed. With decrease in the toxicity of improved pesticides, attention was turned to chronic intoxication and environmental contamination. Nowadays, the problem of diseases not immediately related to the toxic potential of pesticides gains increasing interest. The majority of these non-toxic diseases are dermatoses. Most pesticide-related dermatoses are contact dermatitis, both allergic or irritant. Rare clinical forms also occur, including urticaria, erythema multiforme, ashy dermatosis, parakeratosis variegata, porphyria cutanea tarda, chloracne, skin hypopigmentation, nail and hair disorders. Farmers exposed to arsenic pesticides are at risk of occupational skin cancer, mostly morbus Bowen (carcinoma in situ), multiple basal cell carcinomas and squamous cell carcinomas. Non-arsenic pesticides, e.g. paraquat, are also potentially carcinogenic. PMID- 11426919 TI - Dentist's hand symptoms and high-frequency vibration. AB - The paper discusses characteristics of mechanical vibration as well as the mechanisms of vibration syndrome. Analysis of the relationship between the high frequency vibration and the typical symptoms in the hands of the dentists is presented. Suggestions have been offered on how to limit exposure to vibration in the hands of dentists PMID- 11426920 TI - Risk of exposure to house dust pyroglyphid mites in Poland. AB - During the period of 1989-2000, 335 house dust samples were collected from dwellings at 27 different localities in Poland. Mite allergen exposure was measured in house dust samples collected by performing of mite taxa determination and measuring of the mite allergen levels by a semiquantitative guanine method (Acarex(R) test). Mites were found in 158 of the samples examined (47.2%). A total 3,714 mites were isolated and 15 species identified, including four species from the family Pyroglyphidae (house dust mites). Among them, Dermatophagoides farinae (DF) was predominant (approximately 67% of the total count), followed by D. pteronyssinus (DP) (17.6%) and Euroglyphus maynei (EM) (1.6%). Hirstia chelidonis (HCh) was found for the first time in house dust samples in Poland. DF was predominant in Iwonicz-Zdroj (96.6%), Katowice (91.8%), Sosnowiec (89.4%), Chorzow (94.8%), Bytom (50.9%), Swietochlowice (96.7%) and generally in Upper Silesia (88.2%), whereas DP dominated in Lod (92.9%), Wodzislaw (80.9%), Krakow (45.6%) and Bielsko-Biala (24.8%). Only 14.3% of the mites collected were alive. Total mean number of domestic mites per gram of dust (in all samples examined) was 204.1 +/- 1079.8. The greatest number of mites per 1 gram of dust was 14,971.4. Mite densities and levels of mite allergens (expressed as Acarex test steps) in samples from beds, floors and upholstery furnitures at particular localities in Poland, and in dwellings of atopic versus non-atopic subjects were compared. Highest mite concentrations were usually found in dust from beds, carpets and shutters. D. farinae was distinctly more abundant both per 1 gram of dust and per 1 sample than the species D. pteronyssinus. Other pyroglyphid mites, E. maynei and H. chelidonis, occurred in very small numbers. No significant differences were found between the counts of mites (total and live) in the dwellings of atopic and non-atopic persons. Approximately 49.5% of samples showed positive levels of the mite allergens (Acarex test steps). An influence of some abiotic indoor factors on the mite prevalence in the examined dwellings was analysed separately in relation to samples of bed dust, floor dust and dust from upholstery furnitures. The density of mites was influenced mainly by the type of heating, temperature, type of sleeping accommodation, type of floor or furniture, sampling method, and type of building, whereas levels of the mite allergens were associated with the mite density, relative humidity, month, sampling method, type of building and type of heating. PMID- 11426921 TI - Seroepidemiology of toxoplasmosis in the Lublin region. AB - Reported are results of serologic examinations for the presence of anti Toxoplasma antibodies by direct agglutination in 1,497 people: 1,327 forestry workers and 86 farmers occupationally exposed to T. gondii from the Lublin region (eastern Poland) and 84 inhabitants of the city of Lublin examined as the control group, including 50 blood donors and 34 workers from forestry headquarters. 58.5% positive results in forestry workers, 56.9% in farmers and 46.4% in the control group were obtained. The highest percentages of positive results were obtained in Sosnowica, Wlodawa and Sobibor, all localities in the Chelm district. This finding and the prevalence of clinical cases may suggest that the Chelm district (easternmost area of the Lublin region, bordering Ukraine) is an endemic area of toxoplasmosis. A case of toxoplasmosis in a 39 year old farmer is described in whom reinfection was identified 20 years after primary diagnosis. Rapid increase in specific serologic titres and symptoms typical for toxoplasmosis were noted. The rest of the family and household animals were also found to be positive which supports the suggestion of a family-environmental case of toxoplasmosis. Survey for anti-Toxoplasma antibodies in various domestic and wild animals comprised sera from 262 cows, 120 pigs, 34 geese, 65 chickens, 3 roe deer and 10 sheep from the Lublin region. High percentages of positive results were found in cattle (53.8%) and in pigs (15%). Fowl were positive only in 0-5.9%. The cattle and pigs from the Chelm district are most probably the main sources of toxoplasmosis threatening humans in this area. PMID- 11426922 TI - House dust mites (Acari: Pyroglyphidae) in the cities of Gdansk and Gdynia (northern Poland). AB - 277 samples of house dust collected in 1996-1998 from 17 flats, three hospitals, two hotels and one students' hostel situated in the cities of Gdansk and Gdynia were subjected to acarological examination. Acari were found in 50 (37.3%) of 134 samples from the flats, in 11 (15.5%) of 71 samples from the hospitals and in 13 (18.1%) of 72 samples from the hotels. The majority of mites (91.6%) was found in samples that originated from the private flats. 95.0% of mites from the flats, 35.0% mites from the hotels and 8.0% mites from the hospitals belonged to two dust mite species of the family Pyroglyphidae: Dermatophagoides pteronyssinus and Dermatophagoides farinae. D. farinae was significantly predominant and composed 82.8% of the whole pyroglyphid collection. Samples from private flats contained significantly more mites than those from hospitals and hotels: mean mites densities per 1 gram of dust were 13.07, 1.03 and 1.00, respectively. The authors suggest that only density of house dust mites in private flats may be clinically important. PMID- 11426923 TI - Work-related symptoms in sewage treatment workers. AB - The aim of this study was to investigate health symptoms in sewage treatment workers. A health questionnaire was distributed among 147 sewage treatment workers. Correlating symptoms were clustered using principal component analysis and the association with self-reported exposure was assessed by calculating prevalence odds ratios (OR). Endotoxin was measured in two treatment plants. Personal endotoxin exposure was low (<10 Endotoxin Units/m(3)). Factor analysis of 29 symptoms resulted in four clusters of highly correlating symptoms: 'flu like symptoms'; 'higher airway symptoms'; 'lower airway symptoms'; and 'neurological symptoms'. These clusters were positively associated with working with sewage, but only significant for 'flu- like symptoms' (OR=5.0; 95%CI=1.4 17.6; p<0.05) and 'neurological symptoms' (OR=4.2; 95%CI=1.5-11.7; p<0.01). Chemical exposure was associated with 'neurological symptoms' (OR=8.4; 95%CI=1.1 65.7; p<0.05). The use of daily washed working clothes was negatively associated with 'flu-like symptoms' (OR=0.3; 95%CI=0.1-0.6; p<0.01). In conclusion, sewage treatment workers develop a large variety of work-related symptoms that are not likely caused by endotoxin exposure only. Good hygienic practice at the workplace may prevent some of these symptoms PMID- 11426924 TI - Dermal absorption and distribution of (14)C carbaryl in Wistar rats. AB - The level of (14)C carbaryl was determined in blood (leukocytes, erythrocytes, all blood cells, plasma) and organs (brain, heart, lungs, liver, spleen, skin at the site of exposure) of male Wistar rats after dermal administration. The application liquid was (14)C carbaryl solution in 96% ethyl alcohol. This preparation, possessing an activity of 670 kBq/ml, containing 1.67 mg of carbaryl, was applied to the skin of the tail according to Massmann's method in own modification. The amount of the preparation per 1 cm(2) of the tail skin was 0.19 mg of carbaryl (74.4 kBq). The tails of experimental rats were exposed to (14)C carbaryl by soaking for 4 h daily: once, twice or three times. Beta radiation from (14)C was measured in homogenized organs (brain, heart, lungs, liver, skin) and in blood by computer controlled Wallac scintillation counter Model 1409, using Multi Calc software. The dermal absorption of carbaryl at the site of exposure and in the surrounding area of about 2 cm was observed already during 4 hour exposure. Carbaryl reached plasma within 4 h of a single dermal exposure and penetrated into leukocytes, erythrocytes, heart, liver, lung, kidney and brain. The largest amount of (14)C carbaryl, about 2% of absorbed dose, was detected in liver PMID- 11426925 TI - Work-related skin symptoms and type I allergy among eastern-Polish farmers growing hops and other crops. AB - 73 eastern-Polish farmers growing hops and other crops were examined. They were questioned by a dermatologist and subsequently skin prick-tested with allergens of hops, grain dust, straw dust, hay dust, storage mites, and antigens of microorganisms typical for farm environment. RESULTS: 14 farmers (19.2%) complained of work-related skin symptoms, caused most often by hops (11%), followed by grain (5.6%), hay (5.5%) and straw (4.1%). Five farmers (6.8%) complained of hand dermatitis, four (5.5%) of airborne dermatitis, and eight (11.0%) of pruritus. In two farmers, two skin diseases co-existed. The skin symptoms were mostly mild, however, one case of severe invalidating airborne dermatitis to hops was found. On skin prick tests, 14 farmers (19.2%) showed positive skin reaction to at least one allergen; 5.5% of farmers reacted to grain dust, 5.5% to straw dust, 11% to hay dust, and 8.2% to hops. Tests with storage mites showed positive reactions to Acarus siro in 9.6%, Lepidoglyphus destructor in 17.8%, and to Tyrophagus putrescentiae in 13.7%. Tests with microbial allergens elicited positive reactions to Pantoea agglomerans in 4.1%, Saccharopolyspora rectivirgula in 4.1%, Aspergillus fumigatus in 4.1% and to Streptomyces albus in 1.4% of farmers. Although results of skin prick tests in general did not correlate well with the work- related skin symptoms, in three of 14 farmers with skin symptoms the tests results played a crucial role in identifying the cause of their work-related skin disease. PMID- 11426926 TI - Is there a territorial differentiation in the prevalence of peptic ulcer among rural population in Poland? AB - The aim of this study was to determine the prevalence of peptic ulcer among rural population in various regions of Poland and to analyse the conditions influencing the prevalence of the disease. For organizational reasons, the division of the territory of Poland into eight regions was adopted for the study. The study covered a representative group of 6,512 rural inhabitants, comprising 3,107 males (47.7%) and 3405 women (52.3%), aged 20-64, selected by two-stage stratified sampling. At the first stage of the study all health centres (3,286) were classified into 150 groups and in each group two prevention-treatment regions were selected by means of stratified sampling. The second-stage samples were selected based on communes where the health centres classified for the study were located. People selected for the study were subject to examinations which covered: a specially designed questionnaire form, detailed physical examination, and the necessary specialist tests. The obtained results were recorded in a questionnaire form, which additionally contained questions concerning detailed demographic and social data, hazardous factors present in the working environment, as well as data pertaining to housing conditions, nutrition and habits. Among the rural population under study, peptic ulcer was found in 8.0% of males and 2.9% of females, gastric ulcer was observed in 1.2% of people under study, duodenal ulcer - in 3.2%, gastric and duodenal ulcer - in 0.2%, whereas patients who underwent surgical procedures due to peptic ulcer constituted 0.7% of respondents. Territorial differences were noted in the prevalence of peptic ulcer among Polish rural population. The highest peptic ulcer incidence rates were observed in Macroregion I (western Poland) - where the disease was diagnosed in 7.2% of people under study (Northern Region - 8.1%, Southern Region - 7.4%, and South-Western Region - 6.4%), while the lowest rates were noted in Macroregion II (central and eastern Poland), where peptic ulcer occurred among 4.7% of respondents (South-Eastern Region - 4.4%, North-Eastern Region - 4.5%, Middle-Eastern Region - 4.7%, Middle- Western Region - 4.8%, and Central Region - 5.1%). In regions where the highest incidence rates were noted, the greatest numbers of divorcees, widows and widowers were observed. An analysis by occupational groups showed that in these regions there were more unskilled and skilled workers, employees of services, and the largest number of people performed non-agricultural occupations. Cigarette smoking habit was also more prevalent in these regions. PMID- 11426927 TI - Physical workload on upper extremities in various operations during machine milking. AB - The aim of the study was to quantify the workload on the upper extremity for fundamental work tasks during machine milking. Eleven milkers working in a loose housing system with a milking parlour participated in the study. Muscle activity for the biceps and the forearm flexors, as well as positions and movements of the wrists were simultaneously measured by electromyography and electrogoniometry while video-recording the work. The milking work was broken up in three main tasks "Drying (the cow's udder)", "Pre-milking (the first milk)" and "Attaching (the milking unit to the udder)" and three supplementary tasks. All three main tasks show high muscle load values and almost no time for rest. The highest load values for the biceps and flexor muscles were found during the tasks "Attaching, holding the milking unit" and "Drying", respectively. For 10% of the recording time, the milkers held active hands in 42 degrees dorsal flexion during the milking tasks "Pre-milking" and "Attaching" and in deviated positions exceeding 50% of their maximum values during "Attaching" and "Drying". The high muscle loads in combination with extreme positions and movements of the hand and forearm might contribute to the development of injuries among milkers. The result from the study aims to form a basis for technical improvements of the milking equipment to decrease the risk for arm wrist and hand disorders. PMID- 11426928 TI - Exposure to airborne microorganisms in Polish sawmills. AB - Microbiological air sampling was performed in four sawmills located in eastern Poland, of which two were processing coniferous wood (pine, fir) and other two deciduous wood (oak, birch). Total concentration of microorganisms (bacteria and fungi) in the air of sawmills processing coniferous wood was on average 20.2 +/- 5.6 x 10(3) cfu/m(3) (mean +/- S.E.) and significantly (p < 0.05) higher compared to those processing deciduous wood where the mean concentration of airborne microorganisms was 9.8 +/- 3.0 x 10(3) cfu/m(3). The greatest concentrations of microorganisms in the sawmills processing coniferous wood were noted at debarking and at first-cut frame sawing of pine logs (42.1 +/- 7.6 x 10(3) cfu/m(3) and 39.8 +/- 7.0 x 10(3) cfu/m(3), respectively). Microflora released into air during debarking consisted mostly of allergenic fungi (mainly Aspergillus fumigatus) and corynebacteria, whereas airborne microflora recovered during first-cut frame sawing constituted mostly of endotoxin-producing Gram- negative bacteria of the genus Rahnella, developing in the sapwood of pine. In the sawmills processing deciduous wood, the largest concentration of microorganisms (30.6 +/- 3.4 x 10(3) cfu/m(3)) was found at sorting of the oak parquet boards and was due to the secondary infection of the boards with moulds Penicillium citrinum during prolonged storing in the open air. Values of the respirable fraction of airborne microflora in the examined sawmills varied within fairly wide limits and were between 22.5 86.6%. Altogether, 34 species or genera of bacteria and 21 species or genera of fungi were identified in the air of sawmills, of which respectively 13 and 9 species or genera were reported as having allergenic and/or immunotoxic properties. The concentrations of airborne bacterial endotoxin which were determined on two sampling sites in the sawmills processing pine and fir, were 0.24 microg/m(3) and 4.00 microg/m(3) respectively, distinctly exceeding the suggested safe level. In conclusion, the workers of Polish sawmills may be exposed on some working stands to airborne microorganisms posing respiratory hazard, of which the greatest risk is represented by allergenic fungi developing on bark of logs or stored wood products and endotoxin-producing Gram-negative bacteria of the genus Rahnella, developing in sapwood of coniferous logs PMID- 11426929 TI - Response of sawmill workers to work-related airborne allergens. AB - The aim of this work was to determine the reactivity of sawmill workers to biological allergens associated with wood dust. Allergological examinations by skin and precipitin tests were performed in 43 workers employed in a sawmill processing coniferous wood (pine), in 90 workers employed in two sawmills processing deciduous wood (oak), and in 32 healthy urban dwellers not exposed to organic dusts (referents). The skin test was performed by the intradermal method with the saline extracts of wood dust and of the cultures of three microbial species (Rahnella sp., Brevibacterium linens and Penicillium citrinum) isolated from the air polluted with wood dust. Sawdust from pine was used for testing of the pine processing workers and referents while sawdust from oak was used for testing of the oak processing workers. Skin reactions were recorded after 20 minutes, 8 hours and 24 hours. The agar-gel test for the presence of precipitins in serum was performed with the extract of pine wood dust and extracts of 17 microbial isolates. The workers processing pine showed a very high frequency of positive skin reactions to the extract of wood dust at all time intervals, significantly greater compared to the workers processing oak and referents (p < 0.001). The early skin reactions to the extracts of dust-borne bacteria and fungi were very common among sawmills workers and showed a significant relationship with the degree of exposure. The frequency of reactions to Gram-negative bacterium Rahnella sp. was significantly greater in the pine processing workers than in the oak processing workers and referents (p < 0.001). By contrast, the oak processing workers reacted significantly more frequently to Penicillium citrinum, compared to the pine processing workers and referents (p < 0.01). These results conform to the prior study of airborne microflora in which the dominancy of Gram-negative bacteria was stated in the pine processing sawmill while mould fungi were most common in the oak processing sawmills. The antibody response of sawmill workers to work-related antigens was much weaker compared to skin reactions. As many as 41 sawmill workers reported the occurrence of work-related symptoms. A significant relationship was found between the occurrence of symptoms and frequency of allergic reactions, but only with a limited number of antigens. The obtained results suggest that early allergic reactions to coniferous wood and to microorganisms associated with wood dust are common among sawmill workers, posing a potential risk of work-related disease in this occupational group. PMID- 11426930 TI - Neurological and psychological symptoms after the severe acute neuroborreliosis. AB - The purpose of this study was to delineate distant neurological and neuropsychological effects of severe neuroborreliosis. A group of 33 patients (12 men and 21 women) were selected for the study. Every patient had suffered from severe meningitis, meningoencephalitis or meningopolyradiculoneuritis due to neuroborreliosis in the chronic form of the illness. Standardised medical interview, physical examination and a series of neuropsychological tests (WAIS-R, BDI, BENTON-BENDER, DUM) were performed. In the clinical history, 36.4% of the patients complained of headache, 27.3% of subjective memory distortions; 33.3% of the patients suffered from sleeplessness. The neurological examination showed that 36.4% of the patients experienced such cerebellum integrity disturbances as abnormalities in gait and coordination or even mild ataxia. 21.2% of the patients experienced dysfunction in the proprioceptive pathways, 9% asymmetry in deep tendon reflexes (DTR's), 27.3% disturbances in the sensory responses. The examination showed, however, no muscular strength abnormalities. Half of the patients had slight depression. Psychological tests indicated that 21.2% of the patients had problems in thinking process and experienced memory impairment. 36.4% of the patients had significant organic damage in the central nervous system. The results of this study suggest the existence of long-lasting consequences of acute neuroborreliosis, which can significantly influence the quality of life of patients. PMID- 11426931 TI - Dysarthria as the isolated clinical symptom of borreliosis--a case report. AB - This report presents a case of dysarthria due to hypoglossal nerve mono neuropathy as the only consequence of neuroborreliosis. The 65-year-old man with a seven-months history of articulation disturbances was examined. The speech of the patient was slow and laboured. A slight weakness of the muscles of the tongue (left-side) was observed. The patient suffered from meningitis due to Borrelia burgdorferi infection in 1999 and initially underwent a successful antibiotic treatment. Detailed radiological investigation and psychological tests were performed and co-existing neurological diseases were excluded. To describe profile of speech abnormalities the dysarthria scale was designed based on S. J. Robertson Dysarthria Profile. There were a few disturbances found in self assessment of speech, intelligibility, articulation, and prosody but especially in the morphology of the articulation muscles, diadochokinesis, the reflexes (in the mouth, larynx and pharynx). Needle EMG examination confirmed the diagnosis of mono-neuropathy of left hypoglossal nerve. The study confirms the fact that neuroborreliosis may evoke chronic consequences. PMID- 11426932 TI - Noxious gases in greenhouses. AB - The concentration of NO(2) and SO(2) was measured in a commercial greenhouse from 23/9/1999 25/01/2000. The measurements showed that the level of the two gases is very high in the greenhouse atmosphere. Lung function tests in 42 workers showed that temporary work did not influence significantly the respiratory health status. PMID- 11426933 TI - Evaluation of dental health in mill workers. Part I. The state of dentition. AB - A study of the oral health of workers in flour mills was carried out. The examined group consisted of 40 males and 8 females, currently employed at flour mills. The results of the research indicate the necessity of intensification of stomatological care among mill workers. PMID- 11426934 TI - Intracellular trafficking determinants in APOBEC-1, the catalytic subunit for cytidine to uridine editing of apolipoprotein B mRNA. AB - The posttranscriptional deamination editing of apolipoprotein B (apoB) mRNA catalyzed by APOBEC-1 (apoB mRNA editing catalytic subunit 1) is a nuclear process. The signals in APOBEC-1 responsible for its dual cytoplasmic/nuclear distribution have been evaluated. Residues 97-172 in the middle of APOBEC-1 together with its N-terminal 56 residues affect nuclear localization. Mutagenesis studies however revealed no discrete nuclear localization signal in APOBEC-1. Fifteen amino acids (Leu 173-Leu 187) within the previously identified C-terminal domain of APOBEC-1 were sufficient as a determinant for cytoplasmic distribution in that context. These residues failed to demonstrate nuclear export function in a reporter assay. Further, the distribution of APOBEC-1 in the cytoplasm did not respond to leptomycin B, suggesting that APOBEC-1 did not have nuclear export activity. The data suggested that there are at least three regions in APOBEC-1 that participate in its distribution in both the nucleus and the cytoplasm of editing competent cells; however, none of these meet the functional criteria of nuclear localization or nuclear export signals. The findings are discussed in terms of their implications in the regulation of nuclear editing activity and the possibility that interactions with chaperones may play a role in the cellular distribution of APOBEC-1. PMID- 11426935 TI - Heat-induced proteasomic degradation of HSF1 in serum-starved human fibroblasts aging in vitro. AB - The exposure of human fibroblasts (HF) aging in vitro to heat shock resulted in an attenuated expression of the heat shock-inducible HSP70. When late passage cells were cultured in the continuous presence of serum, we observed a reduced accumulation of the cytoplasmic polyadenylated HSP70 mRNA. The levels of HSF1 activation and nuclear HSP70 mRNA were comparable to those of early passage cells (M. A. Bonelli et al., Exp. Cell Res. 252, 20-32, 1999). When late passage cells were serum-starved overnight, we observed a reduced activation of HSF1 and a decreased level of HSP70 mRNA during heat shock. However, at 37 degrees C the levels of HSF1 differed little between late passage HF and early passage cells, irrespective of the presence of serum. Interestingly, during heat shock a marked decrease in the level and, consequently, in the binding activity of HSF1 was noted only in serum-starved, late passage HF. The decrease in the level of HSF1 was counteracted by back addition of serum to the cells during heat shock. Addition of the specific proteasome inhibitor MG132 blocked a decrease in HSF1 during heat shock, maintaining levels observed in late passage cells and HSF1 activity comparable to that of early passage HF. The recovery of the level and activity of HSF1 observed in late passage HF incubated in the presence of MG132 suggests that heat shock unmasks a latent proteasome activity responsible for HSF1 degradation. PMID- 11426936 TI - Glycolipid-enriched membrane domains are assembled into membrane patches by associating with the actin cytoskeleton. AB - Nonionic detergent lysates of cells contain a glycolipid-enriched membrane (GEM) fraction. It has been proposed that the GEM fraction represents poorly solubilized GEM microdomains, or lipid rafts. However, the properties of GEM domains in intact cells remain controversial. To study the properties of a GEM associated protein using confocal microscopy, GFP was targeted to GEM domains using the N-terminal domain of p56(lck) (LckNT). Imaging of HeLa cells expressing LckNT-GFP showed that it was targeted to large actin-rich patches in the plasma membrane that contained up to a fivefold enrichment of protein. Double-labeling experiments showed that the patches were selectively enriched with other GEM associated molecules. Furthermore, the patches were resistant to extraction by TX 100, and disrupting GEM domains by extracting cholesterol also disrupted colocalization of LckNT-GFP with F-actin. Analogous to the actin-rich patches in HeLa cells, LckNT-GFP colocalized with actin-rich membrane caps in stimulated T cells. Furthermore, disrupting the GEM-targeting signal of LckNT-GFP also inhibited its targeting to membrane caps. Altogether, these findings extend previous studies by showing that association of GEM domains with the actin cytoskeleton provides a mechanism for targeting signaling molecules to membrane patches and caps. PMID- 11426937 TI - Kinetics of efficient recombinant adeno-associated virus transduction in retinal pigment epithelial cells. AB - The aim of this study was to investigate the premise that retinal pigment epithelial (RPE) cells are more permissive to recombinant adeno-associated virus (rAAV) transduction than other cells. We investigated the kinetics and mechanisms of rAAV transduction in RPE cells and found that the transduction efficiencies of cultured RPE cells HRPE51 and ARPE19 were significantly higher than those of 293 (P < 0.008) and HeLa (P < 0.025) cells. In addition, RPE cells reached maximum transduction efficiency at a much lower m.o.i. (m.o.i. 10) than 293 cells (m.o.i. 25). Competition experiments using 1 microg/ml heparin inhibited the high level of transduction in RPE cells by 30%, but additional heparin failed to reduce rAAV transduction further. Southern hybridization of low-molecular-weight DNA from transduced RPE cells indicated that 42% of single-stranded rAAV DNA was translocated into the nucleus by 2 h postinfection. By 6 h postinfection, double stranded rAAV DNA was observed, which coincided with the onset of transgene expression. Southern and fluorescence in situ hybridization of total genomic DNA indicated that long-term transgene expression in RPE cells was maintained by the integration of rAAV into the cellular chromosome. Together, these results suggest that the high permissiveness of RPE cells is not related to the presence of heparan sulfate receptors or nuclear trafficking but may be due to an enhanced rate of second-strand synthesis and that integration in RPE cells is responsible for long-term transgene expression. PMID- 11426938 TI - Stress-inducible transcription factor CHOP/gadd153 induces apoptosis in mammalian cells via p38 kinase-dependent and -independent mechanisms. AB - CHOP/gadd153 is a transcription factor induced by cellular stresses such as UV light, genotoxic agents, and protein misfolding in the endoplasmic reticulum. The fact that these stresses induce CHOP expression, and at the same time cause cellular apoptosis, suggests that CHOP may be directly involved in apoptosis. However, evidence has been circumstantial. Here, we show that CHOP can directly induce apoptosis. A GFP-tagged CHOP vector, ectopically overexpressed in several cell types (3T3 fibroblasts, keratinocytes, and HeLa cells), caused apoptosis as defined by morphology, DNA fragmentation, and FACS analysis. Apoptosis was quantified using a rapid fluorescence assay that measures the signal from cells collected in culture supernatants. The apoptosis-modulating effects of p38 kinase, previously shown to phosphorylate CHOP, were also examined. Simultaneous overexpression of CHOP and p38 significantly augmented apoptosis. However, although p38 kinase clearly modulated the activity of full-length CHOP, it was not absolutely required. Deletion mapping experiments showed that the bZIP region of CHOP stimulates apoptosis to nearly the same extent as wild-type CHOP. Thus, while the amino-terminal region of CHOP serves an important modulatory role (i.e., regulation by p38), the underlying apoptosis-inducing activity of CHOP resides within the bZIP region of the molecule. PMID- 11426939 TI - CD40L induces matrix-metalloproteinase-9 but not tissue inhibitor of metalloproteinases-1 in cervical carcinoma cells: imbalance between NF-kappaB and STAT3 activation. AB - Matrix-metalloproteinases (MMPs) are essentially required for tumor cell invasion and metastasis. Production of precursor enzymes is regulated on transcriptional level, while activation of the pro-enzymes is tightly controlled by posttranscriptional mechanisms. The enzyme activity can be blocked by specific tissue inhibitors of MMPs (TIMPs). In cervical carcinomas strong up-regulation of the type IV collagenase MMP-9 had been demonstrated. We show that activation of CD40, a receptor highly expressed on cervical carcinomas, induces MMP-9 in cervical carcinoma cells, whereas TIMP-1 production inhibiting MMP-9 activity was not affected. This gene induction pattern corresponded to the differential activation of the transcription factor nuclear factor kappaB (NF-kappaB) regulating MMP-9, but not signal transducer and activator of transcription 3 (STAT3), which is involved in TIMP-1 gene regulation. Transient expression of the CD40-inducible NF-kappaB subunit p65 was sufficient for MMP-9 induction. Agents that suppressed CD40-mediated NF-kappaB activation also reduced MMP-9 induction, further supporting an important role of NF-kappaB in CD40-mediated MMP-9 induction. Our data suggest that CD40 expression in carcinoma cells might convert a CD40L-dependent immunological defense signal into a tumor-promoting signal. Selective CD40-mediated signaling through NF-kappaB but not STAT3 correlates to a shift of the balance between MMP-9 and TIMP-1 toward the protease. PMID- 11426940 TI - Human calmodulin-like protein is an epithelial-specific protein regulated during keratinocyte differentiation. AB - Human calmodulin-like protein (CLP) is a calcium-binding protein down-regulated in a cell culture model of mammary tumorigenesis as well as in a majority of breast cancers in vivo. CLP down-regulation may be a result of the poorly differentiated state of these cell lines and tumors, or CLP expression may be incompatible with the uncontrolled cell growth associated with tumorigenesis. To learn more about CLP expression and regulation, we determined the distribution of CLP in various human tissues by immunohistochemistry. CLP was expressed exclusively in the epithelium of the tissues surveyed and was most abundant in thyroid, breast, prostate, kidney, and skin. CLP expression appears to increase in stratified epithelium during differentiation, as illustrated in the skin where CLP staining intensified from the basal through the spinous to the granular layers. Using a normal human keratinocyte culture model, we examined CLP expression in response to various agents known to affect keratinocyte differentiation. Agents that inhibit (epidermal growth factor, EGF) or permit (keratinocyte growth factor) terminal differentiation correspondingly regulate CLP expression. Factors modulating the EGF receptor signaling pathway were particularly potent in regulating CLP expression. CLP expression correlated with an agent's ability to promote terminal differentiation regardless of the agent's effect on keratinocyte proliferation. These studies show that CLP expression is coordinately regulated by, and may be involved in, the program of terminal differentiation in human keratinocytes and, likely, other differentiating epithelial cell types. PMID- 11426941 TI - Characterization of mesonephric cells that migrate into the XY gonad during testis differentiation. AB - In mouse fetal gonads, sex differentiation begins at 10.5-11.5 days postcoitum (dpc). With XY gonads of 12.5 dpc, cord-like structures are visible and stromal cells migrate from adjacent mesonephros, unlike in XX gonads. However, the migrated mesonephric cells, except for the endothelial cells, have not been specifically identified because they have not expressed differentiation markers over the course of organ coculture in previous experiments. In this study, we have for the first time succeeded in isolating only the mesonephric cells that migrate into the XY gonad from the mesonephros with alive and then cultured these cells in vitro through the use of an organ coculture system using EGFP-transgenic mice and a FACS Vantage. The migrated and isolated cells were used for morphological and molecular characterization. The migrated mesonephric cells contained three cell forms; a sharp cell form, a round cell form, and a cluster forming cell. The sharp cells have the characters of peritubular myoid cells. The round cells and cluster-forming cells have the potential to differentiate into Leydig cells, as some of them are 3beta-HSD-positive. In in vitro culture of migrated mesonephric cells, the cluster-forming cells proliferated well and then differentiated into round cells, suggesting that the cluster-forming cells may be stem or precursor cells for the round cells. Thus, our findings provide important information related to the migration and differentiation of migrated mesonephric cells in the XY gonad. PMID- 11426942 TI - Molecular and functional characterization of the four-transmembrane molecule l6 in epidermal keratinocytes. AB - In normal human epidermal keratinocytes (NHEK) proteolytic detachment from the substrate induces a complex activation cascade including expression of new proteins, morphological alterations, and the onset of migration for epidermal regeneration. By subtractive cloning we have shown that L6, a four-transmembrane protein, is newly expressed after proteolytic keratinocyte detachment. In this study, we have generated a novel anti-L6 antibody (clone HD-pKe#104-1.1) and investigated L6 expression regulation in vitro and in vivo as well as L6 function in keratinocyte migration. Dispase-mediated detachment induced L6 expression in NHEK at the mRNA and protein level. Immunohistology of skin biopsies displayed a strong expression of L6 in follicular epidermis and epidermolytic lesions of autoimmune bullous dermatoses (bullous pemphigoid, pemphigus vulgaris), but not in normal interfollicular epidermis. In contrast to normal keratinocytes, HaCaT cells showed constitutive L6 expression, indicating a constitutively active phenotype. After artificial wounding of confluent HaCaT cultures, anti-L6 antibody strongly impaired cell migration velocity and migratory reepithelization of the defect, indicating L6 involvement in keratinocyte migration. These findings suggest that L6 is an important activation-dependent regulator of keratinocyte function and epidermal tissue regeneration. PMID- 11426943 TI - Analysis of apoptosis induced by HIV-1 Vpr and examination of the possible role of the hHR23A protein. AB - The HIV-1 Vpr protein induces apoptosis of cells, the mechanism of which is unknown. To clarify how this function may be related to other Vpr functions, we simultaneously assessed the effects of multiple point mutations upon various Vpr properties. Our data suggest that induction of arrest by Vpr may be unnecessary for induction of apoptosis. This is exemplified by a C-terminal mutant, R80A, that does not arrest cells, yet induces low but significant levels of apoptosis. We also show that mutation of Vpr at both of its nuclear localization sequences (within its alpha-helices and the overlapping leucine zipper-like domain) does not affect induction of either apoptosis or cell cycle arrest. This indicates that neither sequence is essential for these two functions of Vpr. It further suggests that multimerization of Vpr, which maps to residues 60 and 67 within the leucine-rich region, is unnecessary for initiation of apoptosis and arrest. We previously found that the Vpr-binding protein, hHR23A, can partially alleviate induction of arrest. We now show that overexpression of hHR23A itself causes apoptosis of cells. Mutation of its C-terminal UBA( 2 ) domain that is responsible for binding Vpr disrupts the apoptotic effect. This suggests that Vpr may induce apoptosis through a pathway involving hHR23A. PMID- 11426944 TI - HGF/SF induces mesothelial cell migration and proliferation by autocrine and paracrine pathways. AB - Mesothelial repair differs from that of other epithelial-like surfaces as healing does not occur solely by centripetal in-growth of cells as a sheet from the wound margins. Mesothelial cells lose their cell-cell junctions, divide, and adopt a fibroblast-like morphology while scattering across and covering the wound surface. These features are consistent with a cellular response to hepatocyte growth factor/scatter factor (HGF/SF). In this study, we examined the ability of mesothelial cells to secrete HGF/SF and investigated its possible role as an autocrine regulator of mesothelial cell motility and proliferation. We found that human primary mesothelial cells expressed HGF/SF mRNA and secreted active HGF/SF into conditioned medium as determined by ELISA and in a scattering bioassay. These cells also expressed the HGF/SF receptor, Met, as shown by RT-PCR and by Western blot analysis and immunofluorescence. Incubation of mesothelial cells with neutralizing antibodies to HGF/SF decreased cell migration to 25% of controls, whereas addition of HGF/SF disrupted cell-cell junctions and induced scattering and enhanced mesothelial cell migration. Furthermore, HGF/SF showed a small but significant mitogenic effect on all mesothelial cell lines examined. In conclusion, HGF/SF is produced by mesothelial cells and induces both motility and proliferation of these cells. These data are consistent with HGF/SF playing an autocrine role in mesothelial healing. PMID- 11426945 TI - Absence of MyoD increases donor myoblast migration into host muscle. AB - Donor myoblast migration is a major limiting factor in the success of myoblast transfer therapy, a potential treatment for Duchenne muscular dystrophy. A possible strategy to promote the migration of donor myoblasts into host muscle is to enhance their proliferation and delay their fusion, two properties that are major characteristics of myoblasts in regenerating skeletal muscle in MyoD null ( /-) mice. Here we investigate whether the migration of MyoD (-/-) donor myoblasts into host muscle is enhanced in vivo. Sliced muscle grafts from male MyoD (-/-) or normal control (Balb/c) mice were transplanted into the muscles of female normal (Balb/c) host mice. Muscles were sampled at 1, 3, and 12 weeks after grafting, and the fate of male donor myoblasts within female host muscles determined by in situ hybridization with the mouse Y-chromosome-specific Y-1 probe. MyoD (-/-) donor myoblasts migrated into host muscle continuously over 1, 3, and 12 weeks after grafting, in contrast with Balb/c donor myoblasts, whose overall numbers and migratory distances did not increase significantly after 1 week. These results strongly support a role for elevated donor myoblast proliferation and/or their delayed fusion in enhancing migration into host muscle in vivo, and endorse the use of either genetically engineered donor myoblasts, or the administration of exogenous myoblast mitogens to improve donor myoblast migration in myoblast transfer therapy. PMID- 11426946 TI - Respiratory burst responses of rat macrophages to microsporidian spores. AB - This study investigated the respiratory burst responses of rat resident peritoneal macrophages and of peritoneal macrophages stimulated 5 days previously with viable spores of the fish infecting microsporidian Microgemma caulleryi. Nitric oxide production by resident macrophages and prestimulated macrophages in response to viable microsporidian spores was significantly lower than in response to Escherichia coli lipopolysaccharide (LPS) (nitrite concentration in medium 57 +/- 1 microM for resident macrophages stimulated with LPS versus 31 +/- 1 microM for resident macrophages stimulated with microsporidian spores and 36 +/- 4 microM for M. caulleryi prestimulated macrophages; P < 0.05). Extracellular release of reactive oxygen species (ROS) by resident macrophages in response to microsporidian spores was similar to that in response to Kluyveromyces lactis yeast cells and to that in response to phorbol myristate (a stimulator of protein C kinase). Intracellular ROS production by resident macrophages in response to microsporidian spores was similar to that produced in response to yeast cells. Both extracellular ROS production and intracellular ROS production (in response to all stimuli) were significantly lower after in vivo prestimulation of macrophages with microsporidian spores. These results demonstrate that microsporidian spores of species other than those that habitually infect mammals are capable of modulating the respiratory burst of rat peritoneal macrophages. Such modulation may contribute to avoidance by the microsporidian of cytotoxic responses associated with the respiratory burst. PMID- 11426947 TI - Theileria annulata: identification, by differential mRNA display, of modulated host and parasite gene expression in cell lines that are competent or attenuated for differentiation to the merozoite. AB - To identify both host and parasite genes that show altered expression during differentiation of Theileria annulata from the macroschizont to the merozoite stage of the life cycle, the RNA profiles of two T. annulata-infected clonal cell lines (D7 and D7B12) with the same genetic background have been compared by RNA display. In the cloned cell line D7, T. annulata differentiates from the macroschizont to the merozoite at 41 degrees C, whereas in the cell line D7B12, which was derived by recloning D7, the parasite does not differentiate. Therefore, genes that show altered expression levels in either clone could be modulated by the differentiation event and are possible candidates for regulators of this process. Differential display was carried out initially on RNA extracted from D7 and D7B12 macroschizont-infected cells cultured at 37 degrees C and secondly on RNA extracted from the two cell lines incubated at 41 degrees C to induce differentiation to the merozoite. The first procedure identified 29 cDNA fragments that displayed altered levels between D7 and D7B12, 9 of which were confirmed to be differentially expressed by Northern blot analysis. Of these 9 gene fragments, 8 were found to be of host origin, while 1 was parasite derived. The second RNA display analysis identified 14 transcripts that showed altered levels during a differentiation time course, of which 6 were confirmed to be differentially expressed between D7B12 cells and differentiating D7 cells by Northern blot analysis. Of these 6 gene fragments, 1 was of host and 5 were of parasite origin. The parasite genes either showed levels of RNA consistent with constitutive gene expression or, in one case, a genuine upregulation of mRNA associated with the differentiation process. PMID- 11426948 TI - Plasmodium falciparum: a major role for IgG3 in antibody-dependent monocyte mediated cellular inhibition of parasite growth in vitro. AB - In an attempt to identify parasite antigen-specific antibody isotype(s) mediating inhibition of growth in vitro, we tested unfractionated sera and their corresponding purified antibody isotype-containing fractions in in vitro assays with asexual-stage parasites of Plasmodium falciparum in the presence or absence of monocytes. Using affinity purification techniques we fractionated individual and pooled serum samples from semi-immune Gabonese adults, to obtain samples containing either IgG1, 2, 3, and 4, IgG1, 2, and 4, or IgG3 alone, and a non-IgG fraction. Antibodies were quantified spectrophotometrically and the presence of different isotypes in individual fractions was confirmed by protein gel electrophoresis. In the absence of monocytes, we observed inhibition of parasite growth with whole serum and varying levels of either growth enhancement or inhibition with purified Ig-containing fractions. When used in a standardized assay of antibody-dependent cellular inhibition (ADCI) with a monocyte:infected erythrocyte ratio of 1:1, seven of eight serum samples inhibited growth to a mean level of 42%, and the different Ig-containing fractions displayed varying mean levels of inhibition: IgG3, 44%; IgG1--4, 22%; IgG1, 2, and 4, 10%; and non-IgG, 10%. The results suggest that, among the different isotypes present in the serum of semi-immune individuals, parasite antigen-specific IgG3 in particular may play an important role in controlling parasitemia via an ADCI mechanism involving monocyte- derived mediators. PMID- 11426949 TI - Endogenous testosterone levels do not affect filarial worm burdens in mice. AB - It has been observed for several years that males of many mammalian species exhibit higher prevalence and intensity of parasitic infections than females. It has been theorized that this sexual dichotomy may be due to the immunosuppressive properties of testosterone. The administration of exogenous testosterone to laboratory animals often results in higher yields of several parasitic nematodes, lending credence to this hypothesis. We sought to determine if worm burdens in individual mice were related to serum testosterone concentrations. We found that in a cage of five male mice, one or two individuals exhibited high circulating testosterone levels, while the rest had low to undetectable levels. Serum testosterone did not correlate with worm recoveries. The implications of these findings are twofold. First, our data suggest that high endogenous testosterone does not necessarily decrease the ability of mice to resist nematode infection and second, that results obtained from administration of exogenous testosterone should be interpreted with caution. PMID- 11426950 TI - Course of Brugia malayi infection in C57BL/6J NOS2 +/+ and -/- mice. AB - Previous results from our laboratory using pharmacological approaches suggested a role for nitric oxide (NO) in the host defense against the human filarial parasite, Brugia malayi. We sought to determine whether a complementary genetic approach, using mice homozygous for a targeted mutation in the gene encoding inducible nitric oxide-synthase (NOS2), would confirm our observation. We hypothesized that such mice would exhibit some deficit in their ability to clear B. malayi. Our data show that the course of infection in NOS2-/- mice is the same as in wild-type mice. Thus, peritoneal cellular responses to infection are similar in NOS2-/- and wild-type mice, with the exception that T cells form a higher percentage of total peritoneal cells in the former. We find virtually no serum IgE in NOS2-/- mice, suggesting a less robust Th2 response. In contrast, NOS2-/- mice demonstrate an early rise in IgG2a titers compared to B6 +/+ mice. Our data suggest that NO is not an obligate requirement for the elimination of B. malayi from the peritoneal cavities of mice. PMID- 11426951 TI - Role of superoxide and reactive nitrogen intermediates in Rhodnius prolixus (Reduviidae)/Trypanosoma rangeli interactions. AB - This study compares aspects of the superoxide, nitric oxide and prophenoloxidase pathways in Rhodnius prolixus hemolymph, measured in parallel, in response to Trypanosoma rangeli inoculation. Responses to two strains of T. rangeli, and two developmental forms, were studied, and the results obtained were correlated with the ability of the parasites to survive, multiply, and complete their life cycles in the hemolymph of the host. T. rangeli H14 strain parasites, which fail to complete their life cycle in Rhodnius by invading the salivary glands, stimulated high levels of superoxide and prophenoloxidase activity, which peaked 24 h after inoculation. Simultaneously, the concentration of hemolymph nitrites and nitrates increased, indicative of nitric oxide activity, but parasite numbers remained low. T. rangeli Choachi strain parasite inoculation also stimulated superoxide and prophenoloxidase activity, which, though significantly lower than the equivalent responses to the H14 strain, also peaked at 24 h. However, nitrate and nitrite levels in Choachi strain-inoculated hemolymph remained low, and this parasite strain multiplied rapidly, especially following peak superoxide activity, and eventually invaded the salivary glands for transmission to a vertebrate host. In both strains, short form epimastigotes stimulated greater superoxide and prophenoloxidase responses than long form epimastigotes. Injection of the NADPH oxidase inhibitor N-ethylmaleimide or the inducible nitric oxide synthase inhibitor S-methyl isothiourea sulfate caused significantly higher insect mortalities in groups of R. prolixus inoculated with either parasite strain compared with those of uninfected control insects. This indicates that both NADPH oxidase and nitric oxide synthase activity may be involved in the immune response of R. prolixus to infection by T. rangeli. Finally, Western blotting of R. prolixus hemocyte lysates revealed the presence of a protein immunologically related to the human NADPH oxidase complex, the initiator enzyme of the respiratory burst. PMID- 11426952 TI - Lymphangiogenesis and lymph node microdissemination. PMID- 11426953 TI - Abnormal cervicovaginal cytology in women with lupus: a retrospective cohort study. AB - OBJECTIVE: The aim of this study was to review Pap smear reports in women with systemic lupus erythematosus and compare them to a large control population. METHODS: Pap smear results of 29 women with a diagnosis of lupus seen consecutively were compared to those of a control population of 747 women attending the gynecology clinic at the same medical center during the same year. Records of lupus patients were reviewed to obtain clinical data. Fisher's exact test and chi(2) analysis were used to determine statistical significance, as appropriate. RESULTS: Of 29 women with lupus, 1/29 had high-grade squamous intraepithelial lesions (HGSIL) and 6/29 had low-grade squamous intraepithelial lesions (LGSIL). The control population of 747 women had 9/747 with HGSIL and 63/747 with LGSIL. chi(2) and Fisher's exact tests showed that the lupus population had a statistically significant increase in Pap smear reports of dysplasia compared to the control group (P < 0.021 for HGSIL/LGSIL combined, P < 0.036 for LGSIL alone). Examination of serial Pap smear results revealed that 45% of the lupus patients had cervical dysplasia at some time. CONCLUSION: Women with lupus have an increased prevalence of cervical dysplasia. Serial observation revealed dysplastic cytologies in nearly half of the patients, suggesting that this may be a more common problem than previously reported. Serial prospective studies are needed to assess better the risk of premalignant cervical lesions in lupus. PMID- 11426954 TI - Evaluation of recombinant human interleukin-12 in patients with recurrent or refractory ovarian cancer: a gynecologic oncology group study. AB - Objective. The goal of this study was to estimate the antitumor activity and toxicity of recombinant human interleukin-12 (rhIL-12) in patients with recurrent or refractory epithelial ovarian cancer. Methods. From December 1997 to March 1999, patients with recurrent or refractory epithelial ovarian cancer were entered on a Gynecologic Oncology Group phase II study of intravenous rhIL-12. All patients had measurable disease, had a performance status of 0-2, and had failed first-line platinum-based chemotherapy regimen. Eligible patients received rhIL-12, 250 ng/kg IV bolus, as a single dose on Day 1 followed by a 2-week rest period, with subsequent cycles administered daily for 5 days followed by a 16-day rest period per cycle, until disease progression or adverse effects prohibited further therapy. Results. Twenty-eight patients were entered and evaluable for toxicity, while 26 were evaluable for response. The median age was 59.5 years (range: 45-77). The median number of cycles was 2 (range: 1-9). There were no complete responders; however, one patient (3.8%) was a partial responder and 13 patients (50%) had stable disease. Grade 4 myelotoxicity occurred in 21% of patients. Two patients experienced capillary leak syndrome: one grade 2 and one grade 4. Conclusion. As a single agent, rhIL-12 is tolerable and shows a low response rate in recurrent epithelial cancer with measurable disease. PMID- 11426955 TI - Prognostic factors in surgically treated stage ib-iib cervical carcinomas with special emphasis on the importance of tumor volume. AB - OBJECTIVE: The aim of the study was to analyze the importance of tumor volume as a prognostic factor for overall survival (OS) in surgically treated stage Ib-IIb cervical carcinoma. METHODS: One hundred thirteen of one hundred sixty-five patients with histopathological stage Ib-IIb cervical carcinoma (44 Ib1, 24 Ib2, 10 IIa, 35 IIb) treated by radical abdominal hysterectomy between 1989 and 1999, for whom tumor volume could be assessed, were included in this study. Of the 113 patients, 90 (79.6%) received postoperative radiotherapy. Measurement of tumor volume was performed on giant histological sections using a semiautomatic image analyzer. The prognostic significance of tumor volume was analyzed and compared with that of various clinicopathological parameters using uni- and multivariate statistics. RESULTS: The 5-year disease-free survival was 71.4%. Increasing tumor volume was associated with more frequent lymph node metastases and a significant decrease in OS (P = 0.0112). The Median tumor volume was smaller in stage IIa tumors than in stage Ib2 tumors, and histopathological stage did not correlate linearly with lymph node metastases as well as OS. Stage Ib2 tumors were associated with worse overall survival than stage IIa tumors. In univariate analysis, lymph node metastases, histopathological stage, lymph vascular space involvement, tumor volume, parametrial spread, and tumor involvement of resection margins were significant parameters for OS. In multivariate statistical analysis, only lymph node metastases and histopathological staging remained independent prognostic factors for OS. CONCLUSIONS: Tumor volume does not seem to confer additional prognostic information if histopathological stage and lymph node status are known. However, it may provide important prognostic information if lymph node status is not known or histopathological stage cannot be assessed. PMID- 11426956 TI - Positive emission tomography for evaluating a complete clinical response in patients with ovarian or peritoneal carcinoma: correlation with second-look laparotomy. AB - OBJECTIVE: Positive emission tomography (PET) provides a novel means of imaging malignancies. The following study was undertaken to evaluate the predictive value of PET in determining a pathologic complete response in patients with advanced ovarian or peritoneal carcinoma who had a complete clinical response following primary chemotherapy. METHODS: Twenty-two patients with advanced-stage ovarian (N = 17) or peritoneal (N = 5) carcinoma who had achieved complete clinical and radiologic remission and normal CA-125 level after six cycles of chemotherapy and who had consented to a second look laparotomy procedure were studied. All patients received platinum based therapy and all but one patient, treated elsewhere, received paclitaxel in combination with platinum. Following IV administration of 20 mCi [(18)F]fluorodeoxyglucose (FDG), the entire abdomen and pelvis were scanned. Various technical modifications including bladder activity dilution, intravenous hydration with diuretic therapy, and mechanical bowel preparations, were used to reduce background activity. Second-look laparotomy findings were classified as negative, macroscopically positive if a biopsy of a suspicious area was histologically positive, or microscopically positive if only a nonsuspicious area was histologically positive. The effect of patient preparation prior to PET imaging was evaluated. RESULTS: Persistent disease was found in 13 of the 22 patients (59%). Only one of nine sites with macroscopic and none of four with microscopic disease were accurately predicted. The sensitivity was only 10% and the specificity 42%. Intravenous hydration, diuretic therapy, and bowel preparation did not improve the results. CONCLUSIONS: These results suggest that despite technical modifications the sensitivity of PET before second look laparotomy for small-volume persistent disease is low. PMID- 11426957 TI - The correlation between colposcopically directed cervical biopsy and loop electrosurgical excision procedure pathology and the effect of time on that agreement. AB - OBJECTIVE: The goal of this study was to evaluate whether colposcopically directed cervical biopsy accurately predicts histopathology found on loop electrosurgical excision procedure specimen and to assess whether this correlation was affected by a delay in treatment of greater than 12 weeks. METHODS: A retrospective review was performed of all cervical biopsy and loop electrosurgical excision procedure pairs performed at University of Arizona Medical Center between March 1992 and March 2000. Agreement was assessed by kappa statistics and Spearman rho coefficients. A subgroup analysis was performed to assess the effect of a delay greater than 12 weeks between cervical biopsy and loop excision on the histopathologic correlation. RESULTS: An 84% agreement was found between cervical biopsy histopathology and loop electrosurgical excision procedure histopathology when compared to within 1 degree (kappa = 0.78, P < 0.0001; Spearman rho = 0.40, P < 0.0001). This agreement remained strong (87%) even when loop electrosurgical excision procedure was delayed greater than 12 weeks (kappa = 0.81, P = 0.01). CONCLUSION: Colposcopically directed cervical biopsy correlated strongly with loop specimen histopathology. A delay in loop electrosurgical excision procedure greater than 12 weeks does not negatively affect this correlation. This delay of 12 weeks may be used safely in the future to test chemotherapeutic dysplasia treatments. PMID- 11426958 TI - High prevalence of oncogenic human papillomavirus in the genital tract of women with human immunodeficiency virus. AB - OBJECTIVE: The goal of this study was to determine the prevalence of human papillomavirus (HPV) and squamous intraepithelial lesions (SILs) in women infected with human immunodeficiency virus (HIV) in Mexico. METHODS: Cases included women who were positive for human immunodeficiency virus (HIV) and accepted to participate. There were two control groups in this study: group A, heterosexual partners of HIV+ men; group B, commercial sex workers. Gynecologic examination was performed in all participants. Also, a cervical smear with colposcopy and a sample for detection of HPV DNA by polymerase chain reaction (PCR) were obtained in all subjects, as were CD4+ counts. Relative risks (RR) and 95% confidence interval were calculated. RESULTS: Eighty-five HIV+ women agreed to participate in this study; the route of HIV infection was heterosexual in 78.8%; transfusion in 8.2%; paid donors in 3.5%; and 9.4% unknown. A total of 9 controls were included: 4 from group A and 5 from group B. HPV DNA was detected by PCR in 57 (69%) cases and in 26 (29%) controls from both groups (P < 0.0001). The RR of HPV infection was 5.5 (2.7-11.5). Also, a significant difference in the prevalence of high-risk HPV types was observed between cases and controls, RR = 12.8 (4.07-42.9). These associations were independent of CD4+ counts and antiretroviral therapy. No association was observed between HIV infection and the risk for high-grade SIL. CONCLUSIONS: We observed a high prevalence of oncogenic HPV types in HIV-positive women. These women should be screened regularly for early diagnosis of premalignant lesions and prevention of cervical cancer. PMID- 11426959 TI - Neuroendocrine cervical carcinoma: a diagnostic and therapeutic challenge. AB - OBJECTIVE: The aim of this study was to analyze diagnostic criteria, response to chemotherapy, rate and site of relapse, and overall survival (OS) in neuroendocrine cervical carcinoma. METHODS: Twenty patients were included. Stage was Ia(2) in 1 case, Ib(1) in 4, Ib(2) in 4, II in 5, IIIb in 2, IVa in 2, and IVb in 2. Patients with stage Ib(2) or greater received neoadjuvant chemotherapy (NCH). Eighteen patients were operated on. Immunohistochemistry was performed on the surgical specimens. Statistical analysis included the Kaplan-Meier method and the chi(2) and log-rank tests. RESULTS: The response to NCH was <50% in 2/13 cases (15.3%), >50% in 9/13 (69.4%), and complete in 2/13 (15.3%). Cytokeratin was positive in 17/18 cases, neuron-specific enolase in 15/18, chromogranin in 9/18, and synaptophysin in 8/18. Tumor was pure in 12 cases. Two cases had simultaneous ovarian carcinoma. Positive nodes were observed in 9/20 pts (45%). Tumors <4 cm had no recurrences. Pure tumors >4 cm had distant relapses (6/11). Mixed tumors >4 cm had 2/6 pelvic and 3/6 lung metastases. OS was 39%. When the initial tumor volume was <4 cm OS was 76%, and it was 18% for tumors >4 cm (P < 0.05). OS was 58% when the residual tumor after NCH was <2 cm and 21% when it was >2 cm (P < 0.05). When the tumor was pure OS was 54% and 19% when it was mixed (P < 0.05). OS was 72% among pts with negative nodes and 11% for those with positive nodes (P < 0.01). CONCLUSIONS: (1) Stage IV was frequent (20%); (2) Response to NCH was high; (3) The pattern of relapse differs for mixed tumors; (4) For tumors <4 cm outcome is similar to that of squamous carcinoma. PMID- 11426960 TI - Three-dimensional power Doppler ultrasound improves the diagnostic accuracy for ovarian cancer prediction. AB - OBJECTIVE: The goal of this study was to determine if three-dimensional power Doppler ultrasound improves the specificity for ovarian cancer detection as compared with two-dimensional ultrasound. METHODS: Seventy-one women with a known complex pelvic mass were referred for a preoperative ultrasound evaluation with both two-dimensional and three-dimensional gray-scale ultrasonography. The 3D studies were performed with the Kretz Voluson 530D using a mechanized transvaginal probe. Surface rendering and power Doppler imaging were performed by the same gynecologic sonologist, and reassigned to one of four echo patterns: cystic, multicystic, complex, or solid. Sonographic criteria used for diagnosing ovarian cancer were based on a system that included morphological characteristics, histological prediction, and power Doppler imaging. RESULTS: Seventy-one women underwent surgical exploration: 14 (19.7%) had ovarian cancer (2 FIGO stage I, 2 stage II, 7 stage III, and 3 metastatic colon) and 2 had uterine cancer. Two-dimensional gray-scale ultrasound identified 40 masses as suspicious for cancer, including all 14 malignancies, yielding a sensitivity, specificity, and positive predictive value of 100, 54, and 35%, respectively. However, evaluation with 3D power Doppler identified only 28 cases as suspicious (including all 14 cancers), resulting in a sensitivity, specificity, and positive predictive value of 100, 75, and 50%, respectively. CONCLUSIONS: Three dimensional power Doppler imaging better defines the morphological and vascular characteristics of ovarian lesions. All malignancies were correctly identified by both 2D and 3D imaging; however, the specificity significantly improved with the addition of 3D power Doppler. This improved diagnostic accuracy may promote improved patient care by separating complex benign masses from ovarian cancer, therefore facilitating appropriate physician referral. PMID- 11426961 TI - Interleukin-6 in cervical cancer: the relationship with vascular endothelial growth factor. AB - OBJECTIVE: Interleukin-6 (IL-6), a central proinflammatory cytokine, has been implicated in cervical cancer, though its role remains elusive. This study was an attempt to elucidate the role of IL-6 in the pathogenesis of cervical cancer, with particular emphasis on tumor angiogenesis. METHODS: Cytosolic IL-6, vascular endothelial growth factor (VEGF), and platelet-derived growth factor (PDGF) levels were determined via enzyme immunoassay in 60 FIGO stage IB-IIA cervical cancer patients. Immunohistochemical staining in tissue sections was performed to analyze the distributions of IL-6 and IL-6 receptors. Meanwhile, human papillomavirus (HPV) DNA was detected by polymerase chain reaction-based survey. In vitro studies of two cervical cancer cell lines, C33A and SiHa, for the interaction between IL-6 and VEGF were also performed. RESULTS: Consistently higher expression of IL-6 and VEGF was evident in cancerous tissues than in adjacent noncancer tissues in early-stage cervical cancer patients (P < 0.01). After recombinant human IL-6 was added, VEGF was induced in a time- and dose dependent manner in cervical cancer cell line C33A. Correspondingly, interrupting the IL-6 autocrine machinery with either anti-IL-6 or anti-IL-6 receptor antibody markedly reduced the expression of VEGF at the transcriptional level in SiHa cells. Significantly higher levels of IL-6 in cancer tissues were observed in patients older than 45 (P < 0.01), patients with tumors >2 cm (P < 0.01), patients with oncogenic HPV-16 or -18 infections (P < 0.01), and patients with squamous cell carcinoma (P = 0.02). Patients with a deeper stromal invasion, vaginal invasion, lymphovascular emboli, or lymph node metastasis appeared to have higher intratumoral IL-6 levels, although the differences were statistically insignificant. CONCLUSIONS: Substantially high microenvironmental IL-6 levels promote tumor angiogenesis and the development of cervical cancer. Thus, inhibition of the biological activity of IL-6 may be potentially beneficial. PMID- 11426962 TI - An immunomagnetic-based method for the purification of ovarian cancer cells from patient-derived ascites. AB - OBJECTIVE: Primary ovarian cancer cells obtained from fresh tumor have many advantages over established cell lines. Therefore, a procedure for the specific and efficient purification of such neoplastic cells is critical. We report an effective immunomagnetic method for the isolation of tumor cells from the ascitic fluid of patients diagnosed with ovarian adenocarcinoma. METHODS: This procedure incorporates the use of monoclonal antibody (mAb) CC49, which recognizes the tumor-associated glycoprotein 72 (TAG-72). TAG-72 is highly expressed on ovarian tumor cell surfaces with little or no reactivity with normal tissues. Also used in this protocol are immunomagnetic beads, which bind to the CC49 mAb via a secondary antibody. When ovarian cancer cells adhere to the magnetic beads, a magnetic field is used to separate the tumor cells from all other cellular components. RESULTS: Using ascitic fluid from five patients, we found that preparations before purification contained between 38 and 52% neoplastic cells. Using our method, we produced preparations that were between 63 and 96% pure for cancer cells, thus obtaining an average increase in tumor cell enrichment of 86%. CONCLUSION: We, therefore, believe this method is preferable for producing high yields of pure ovarian neoplastic cells. We are now employing this technique in our laboratory to provide a stringent and pure template for our studies on gene transfer to primary ovarian cancer cells. PMID- 11426963 TI - Characteristics of ovarian and uterine cancers in a cohort of in vitro fertilization patients. AB - OBJECTIVE: The aim of this study was to describe the features of cancers of the uterus, ovary, and other uterine adnexae in a large cohort of in vitro fertilization (IVF) patients and to thereby assist gynecologic oncologists in counseling patients with a history of infertility. METHODS: Record-linkage to population-based cancer registries was used to determine the incidence of invasive cancer in a cohort of 29,700 IVF patients. Pathology reports were made available by the cancer registries and reviewed blind to IVF treatment status. Patient information was abstracted from IVF clinic records. RESULTS: Twelve cancers of the uterus and 13 cancers of the ovary and other uterine adnexae were identified and pathology reports were reviewed for all but 1. Patients ranged in age at diagnosis from 28 to 48 years. Five cancers were diagnosed within 8 months of referral for IVF or IVF treatment; the others were diagnosed between 1 and 12 years later. The ovarian cancers were of the following histologic types: serous (4), mucinous (1), seromucinous (1), endometrioid (3), clear cell (2), and unknown type (1). A choriocarcinoma of the fallopian tube was diagnosed in a woman with a history of tubal infertility. Cancers of the uterus included endometrial adenocarcinomas (8), stromal sarcomas (2), and leiomyosarcomas (2). Ovarian cancer was diagnosed in a woman with Bloom's syndrome, a rare autosomal recessive disorder associated with increased cancer risk. CONCLUSION: Cancer cases in this large, well-defined cohort of IVF patients show a broad range of patient and tumor characteristics. We found an increased incidence of uterine sarcoma in women with a history of infertility. Increased ascertainment of preexisting cancers in the course of infertility investigations and management is to be expected. PMID- 11426964 TI - Allelotype of papillary serous peritoneal carcinomas. AB - OBJECTIVE: Papillary serous peritoneal carcinoma (PSPC) is histologically indistinguishable from papillary serous ovarian carcinoma (PSOC) with a similar clinical presentation, yet may differ in its carcinogenesis. The purpose of this study was to determine the incidence of allelic loss and the frequency of p53 mutation by p53 overexpression in PSPC compared to PSOC. METHODS: An allelotype analysis of 26 patients with PSPC was performed using 39 microsatellite markers from 25 chromosomal arms. Thirty-seven previously studied patients with PSOC served as the comparison. P53 mutations were detected by immunohistochemical protein overexpression. RESULTS: There was significantly less LOH in PSPC than PSOC. Both the number of chromosomes with LOH and the proportion of tumors with allelic loss were less frequent. Significant LOH, defined as >/=30% of informative tumors having loss at a chromosome locus, was seen on 4 chromosome arms in PSPC: 12p, 17p, 17q, and 18q, compared to 18 arms in PSOC: 4q, 5q, 6p, 6q, 9p, 9q, 12p, 12q, 13q, 15q, 16q, 17p, 17q, 18q, 19p, 19q, 22q, and Xq (P < 0.001). The median LOH frequency was higher in PSOC than PSPC, 43% versus 33%, respectively (P = 0.013), and more PSOC tumors had LOH than PSPC tumors, 91% versus 65% (P = 0.042). P53 overexpression was detected in 80% of PSPC tumors. CONCLUSIONS: LOH occurs less frequently in PSPC compared to PSOC. Chromosomal regions with high frequencies of LOH common to PSPC and PSOC, such as 12p, 17p, 17q, and 18q, may harbor tumor suppressor genes important in the carcinogenesis of both malignancies and likely include p53. PMID- 11426965 TI - Synthetic peptides induce a cytotoxic response against human papillomavirus type 18. AB - OBJECTIVES: Over 90% of cervical carcinomas express human papillomavirus (HPV) E6 and E7 proteins. These unique antigens are ideal targets for the development of cytotoxic T-lymphocytes (CTL) for antitumor immunotherapy. In this study we identify peptides from HPV-18 E6 and E7 proteins that bind to HLA class I molecules. We further show that these peptides are able to induce peptide specific CTL from an HLA-A2-positive (+) peripheral blood donor in vitro. METHODS: A computer-assisted algorithm was devised to identify peptides from HPV 18 E6 and E7 proteins that bind to HLA-A2 molecules. Peptides that were predicted to bind were synthesized and their binding activity was determined. HLA-A2(+) irradiated stimulator cells pulsed with HPV-18 peptides were incubated with HLA A2(+) peripheral blood mononuclear cells. Cytotoxicity assays were performed to assess specific cell lysis. RESULTS: Of 295 possible sequences, the computer assisted algorithm predicted 10 peptides that would have a high probability of binding to HLA-A2. The 4 strongest binding peptides were analyzed for their ability to induce cytotoxic cells against HPV-18 peptide-pulsed targets. Two of the peptides induced significant lysis. CONCLUSIONS: There are limited data on peptide-based immunotherapy for HPV-18(+) tumors. The combination of our computer assisted algorithm and binding assay permits rapid selection of potential CTL epitopes. We identified two peptides that were able to induce peptide-specific lysis. These two epitopes are candidates for a peptide-based vaccine against HPV 18(+) tumors. The model described has broad applications and can be used in the development of immunotherapy for other types of cancers. PMID- 11426966 TI - Uptake of a cholesterol-rich emulsion by neoplastic ovarian tissues. AB - OBJECTIVE: Previously, it was shown that a lipidic emulsion (LDE) composed of phospholipids and cholesterol esters which binds to low-density lipoprotein (LDL) receptors may concentrate in acute myeloid leukemia cells. In this study, we aimed to verify whether LDE also has the ability to concentrate in malignant ovarian cancer after being injected into the blood circulation of the patients. METHODS: Three groups of women scheduled for surgery were included in the survey: 13 bearing malignant tumors, 9 with benign ovarian tumors, and 13 without ovarian tumor who were scheduled to undergo oophorectomy due to malignant disease of the uterine cervix or endometrium. On the day prior to surgery they were injected with LDE labeled with [(14)C]cholesteryl oleate. Specimens of tumors and normal ovaries excised during surgery were lipid extracted and analyzed for radioactivity counting. Results were expressed in radioactive count (cpm) per gram of tissue. RESULTS: The mean of the uptakes of the emulsion radioactivity by the malignant tumors was roughly eightfold greater when compared with that of the contralateral normal ovaries (2261 +/- 1444 and 275 +/- 137 cpm/g, respectively, P < 0.012), benign tumors, and normal ovaries of the patients without ovarian tumors. CONCLUSION: LDE has the ability to concentrate in malignant ovarian tumor tissue. Therefore, it can be used as a vehicle to direct cytotoxic drugs against malignant ovarian tumors, thus diminishing the side effects of chemotherapy. PMID- 11426967 TI - Ten-year survival of patients with locally advanced, stage ib-iib cervical cancer after neoadjuvant chemotherapy and radical hysterectomy. AB - OBJECTIVE(S): The aim of this study was to evaluate the effects of neoadjuvant chemotherapy and radical hysterectomy on long-term survival in stage IB-IIB locally advanced cervical cancer by conducting a 10-year follow-up. METHODS: Between August 1983 and May 1990, 80 locally advanced, stage IB-IIB cervical cancer patients with tumor diameter greater than or equal to 4 cm were treated with neoadjuvant VBP chemotherapy (cisplatin, vinblastine, and bleomycin) followed by radical hysterectomy with pelvic lymphadenectomy. After this therapeutic modality, patients were followed for more than 10 years. Ten-year survival rates and factors affecting recurrence after this therapy were evaluated. RESULTS: Of 80 patients, 75 (93.7%) showed a reduction in tumor size after neoadjuvant chemotherapy. At pathologic examination, stage reduction was noted in 53 (66.2%) patients and 20 patients (25%) showed no residual or microinvasive cervical tumor. Pelvic lymph node metastases were found in 17 patients (21.3%). During the 10-year follow up, 2 patients were lost and 16 patients recurred. Overall 5-year and 10-year disease-free actual survival rates were 82.0 (64/78) and 79.4% (62/78), respectively. Clinical stage, initial tumor size, clinical response, and residual tumor size were not risk factors for recurrence after this therapy. However, pelvic lymph node metastasis was a significant risk factor for recurrence. CONCLUSION(S): Neoadjuvant VBP chemotherapy followed by radical hysterectomy in locally advanced, stage IB-IIB cervical cancer patients seemed to improve the long-term survival rate for these patients compared to that of conventional therapy. However, randomized controlled trials are needed to confirm this result. PMID- 11426968 TI - Allele loss on chromosome 1p36 in epithelial ovarian cancers. AB - OBJECTIVES: Prior studies have shown that allelic loss on chromosome 1p36 occurs frequently in ovarian as well as several other types of cancer. This suggests that inactivation of gene(s) in this region may play a role in the pathogenesis of these cancers. The aim of this study was to further delineate the region of loss on chromosome 1p36 in ovarian cancers and to identify associated patient or tumor characteristics. METHODS: Paired normal/cancer DNA samples from 75 ovarian cancers (21 early stage I/II and 54 advanced stage III/IV) were analyzed using microsatellite markers. RESULTS: Forty-nine of 75 (65%) ovarian cancers had loss of at least one marker. The marker demonstrating the most frequent loss was D1S1597, which was lost in 29/57 (51%) informative cases. Allele loss on 1p36 was significantly more common in poorly differentiated ovarian cancers (73%) relative to well or moderately differentiated cases (48%) (P = 0.03). Evidence was obtained for two common regions of deletion: one flanked by D1S1646/D1S244 and another more proximally by D1S244/D1S228. CONCLUSION: These findings further delineate regions on chromosome 1p36 proposed to contain tumor suppressor gene(s) that may play a role in the development and/or progression of epithelial ovarian carcinoma. Allele loss on 1p36 is associated with poor histologic grade. PMID- 11426969 TI - Topoisomerase II immunostaining as a prognostic marker for survival in ovarian cancer. AB - OBJECTIVE: This study aimed to evaluate topoisomerase II compared to Ki-67 expression as a marker for tumor behavior and for prognosis of patients with ovarian cancer. METHODS: In order to screen for potential prognostic markers, two groups of patients with advanced stage (FIGO stages III and IV) epithelial ovarian carcinoma were selected based on differences in survival (mean survival, 11 years versus 2 years). Pathology slides were reviewed, and immunohistochemistry using antibodies to topoisomerase II and Ki-67 was performed on the original cell blocks. No patients were lost to follow-up. RESULTS: Detectable expression of topoisomerase II was present in 70.0 +/- 30.3% of cells in patients with rapidly progressing disease, compared to only 12.3 +/- 12.4% of cells in long-term survivors (P = 0.0001). Ki-67 expression was also more frequent in short-term survivors compared to long-term survivors, but the difference was less prominent than with topoisomerase II (P = 0.01). Specificity and sensitivity as prognostic factors reached 88.2 and 93.8% for topoisomerase II, compared to 55.6 and 88.2% for Ki-67. CONCLUSIONS: Topoisomerase II expression as detected by immunohistochemistry in tumor samples emerged as a promising clinically relevant biomarker for survival in advanced epithelial ovarian cancer. PMID- 11426970 TI - Prognostic factors of Krukenberg's tumor. AB - OBJECTIVE: The aim of this study was to determine prognostic factors of metachronous Krukenberg's tumors of gastric origin, thereby helping to establish a therapeutic plan for this rare entity. METHODS: Thirty-four female patients who underwent curative resection of gastric carcinoma from 1987 through 1996 and subsequently developed Krukenberg's tumors were identified. The covariates used for survival analysis were patient age at the time of ovarian relapse (categorized as <50 years or >/=50 years), size of Krukenberg's tumor (categorized as 10 cm), initial AJCC stage of gastric carcinoma, relapse-free interval (categorized as <1, 1-2, or >/=2 years), extent of recurrent disease, and the presence of gross residual disease after treatment for Krukenberg's tumors. The influence of all covariates on the survival period was assessed using the Cox proportional hazards model. RESULTS: At a median follow-up of 32.3 months (range, 2.2-83.1), the median survival period of 34 study patients was estimated to be 7.7 months (95% confidence interval (CI), 5.8-9.6) after the diagnosis of metachronous Krukenberg's tumor. Cox-model-based survival analysis demonstrated that the absence of residual disease after treatment and limited disease extent were favorable prognostic factors. The median survival period of patients without gross residual disease was longer (10.9 months (95% CI, 4.6 17.1)) than that of patients with gross residual disease (7.5 months (95% CI, 6.6 8.3)) [P = 0.036]. Median survival periods according to the extent of metastasis were 10.9 months (95% CI, 0-23.0) for patients with disease confined to the ovaries, 13.1 months (95% CI, 5.9-20.3) for patients with disease confined to the pelvis, 7.5 months (95% CI, 6.8-8.1) for patients with intraabdominal disease, and 3.6 months (95% CI, 3.4-3.7) for patients with disease spread outside the abdomen and pelvis [P = 0.003]. In contrast, patient age, size of ovarian tumor, initial stage of gastric adenocarcinoma, and relapse-free interval were not prognostic indicators for survival after the development of ovarian metastases. CONCLUSION: The absence of residual disease after treatment and limited disease extent were favorable prognostic factors of metachronous Krukenberg's tumors of stomach origin. Our data suggest that resection might have a role in the management of Krukenberg's tumors of stomach origin if it could render patients free of gross residual disease. PMID- 11426971 TI - Carboxyterminal telopeptide of type I collagen (ICTP) in predicting prognosis in epithelial ovarian cancer. AB - OBJECTIVE: The aim of this study was to assess the prognostic value of serum carboxyterminal telopeptide of type I collagen (ICTP) in ovarian cancer. Serum CA125 was used as a reference marker. METHODS: Forty-five patients with epithelial ovarian cancer were monitored with serial measurements of serum concentrations of ICTP, a degradation product of type I collagen likely to come about via the matrix metalloproteinase pathway. RESULTS: The patients with a good prognosis had significantly lower serum ICTP concentrations than the patients with a poor prognosis both before the operation and at all the postoperative time points studied (3, 6, 9, 12, 18, and 24 months), whereas a corresponding difference in CA125 was first seen after a 12-month follow-up. In multivariate regression analysis, the 9-month serum ICTP concentration remained the only independent prognostic indicator of all biochemical, clinical, and histological variables. The postoperative serum ICTP concentration did not correlate with the clinical stage, the grade of differentiation, or the presence of residual tumor. In contrast to ICTP, postoperative serum CA125 correlated with the clinical stage and the presence of residual tumor. CONCLUSIONS: Because our ICTP test does not detect defectively cross-linked carboxyterminal telopeptides of type I collagen, which is the predominant form in malignant ovarian tissue, the excess ICTP of ovarian cancer patients must originate from the tissue around the tumor, where the malignancy is causing tissue damage. As an indicator of invasion, the serum ICTP test opens up new possibilities to assess the clinical behavior of ovarian cancer and, in the future, also the effect of possible antiproteinase treatment in ovarian cancer. PMID- 11426972 TI - Production of steroids by human ovarian surface epithelial cells in culture: possible role of progesterone as growth inhibitor. AB - OBJECTIVE: The purpose was to investigate whether normal ovarian surface epithelial cells, harvested from premenopausal and postmenopausal women, are capable of steroid production, and to evaluate effects of estradiol and progesterone on growth regulation of such cells. METHODS: Ovarian surface epithelial cells were obtained by brushing of the ovarian surface of 9 premenopausal and 10 postmenopausal women undergoing surgery for benign gynecological diseases. The conditioned media after culture, with and without addition of FSH and LH, were analyzed for estradiol and progesterone. The proliferative effects of the steroids were analyzed using two different culture models, nonconfluent cells and confluent cells, and two different detection methods, [(3)H]thymidine incorporation and a colorimetric method assaying cell number. RESULTS: The normal ovarian surface epithelial cells were found to secrete both estradiol and progesterone, a production that was not regulated by FSH or LH. Addition of steroids to the cultured cells did not induce any overall significant growth effects. However, progesterone significantly inhibited the growth of ovarian surface epithelial cells from three of the patients. Enhanced thymidine incorporation was observed in the presence of the progesterone receptor antagonist Org 31710 in the nonconfluent cultures of cells from postmenopausal women, but no effect of an estrogen receptor antagonist was observed. CONCLUSIONS: The normal ovarian surface epithelium is capable of steroid production, which is also often observed in tissue from ovarian epithelial tumors. Progesterone appeared to be a negative regulator of ovarian surface epithelial growth, while estradiol had no effect. PMID- 11426973 TI - The effect of nulliparity on survival in endometrial cancer at different ages. AB - OBJECTIVE: Nulliparity is a major independent risk factor for endometrial cancer in Japan. We examined the effect of nulliparity on survival in endometrial cancer at different ages. METHODS: A retrospective study of 328 Japanese women with endometrial cancer was performed. The subjects were divided into two groups: a younger age group (women < 50 years) and an older age group (women >/= 50 years). Parity was analyzed for its influence on survival. RESULTS: No effect of nulliparity was observed on survival in the younger group. In the older women, nulliparity did not affect survival in 189 subjects with surgical stage I and II tumors (P < 0.27). In contrast, the cumulative 10-year survival rates associated with nulliparity, a parity of 1 or 2, and a parity of 3 or more were 7.7, 48.0, and 56.2% in 54 subjects with surgical stage III and IV tumors, respectively (P < 0.03). In these 54 subjects, the cumulative 10-year survival rates associated with < 6-month and > 7-month delays in diagnosis were 57.1 and 16.6%, respectively (P < 0.02). The prognostic impact of parity disappeared after adjustment for delay in diagnosis. Multivariate analysis including histopathological variables, parity, and delay in diagnosis showed no independent prognostic variable in the older subjects with surgical stage III and IV tumors. CONCLUSIONS: The negative effect of nulliparity on survival was observed in the older subjects with advanced-surgical-stage tumors. Delay in diagnosis contributed to the prognostic impact of nulliparity. PMID- 11426974 TI - Inhibitory effects of 17beta-estradiol and progesterone on ovarian carcinoma cell proliferation: a potential role for inducible nitric oxide synthase. AB - OBJECTIVES: Indirect evidence suggests that estrogen and progesterone are involved in the etiology of ovarian cancer (Oca). Estrogen and progesterone are also thought to modulate nitric oxide (NO) in human ovarian tumor tissue via regulation of inducible nitric oxide synthase (iNOS). Objectives in this study were: (1) to investigate the effects of 17beta-estradiol (E(2)) and progesterone (P(4)) on Oca cell proliferation employing elevated hormone concentrations occurring within the microenvironment of the ovary, and (2) to determine whether E(2) or P(4) affects iNOS expression and NO generation in Oca cells. METHODS: Proliferation assays assessed the effects of E(2) and P(4) on cell growth in three human Oca cell lines (HOC-7, OVCAR-3, SKOV-3). Reverse transcriptase polymerase chain reaction was used to assess mRNA expression and Western blots to determine protein levels. NO generation was determined via the Griess reaction. RESULTS: Elevated E(2), P(4), or E(2) plus P(4) (E + P), significantly inhibited HOC-7 cells and OVCAR-3 cells, but not SKOV-3 cells. E(2) at 10 microM downregulated iNOS expression and significantly reduced NO production in HOC-7 cells, while 10 microM P(4) or 10 microM E + P increased iNOS expression and NO production. Conclusions. Our findings demonstrate that elevated E(2), P(4), or E + P results in significant growth inhibition of Oca cells, and we propose a role for iNOS and NO in how these hormones modulate their activities in Oca cells. PMID- 11426975 TI - Evaluation of a region on chromosome 1p in ovarian serous carcinoma that is frequently deleted in uterine papillary serous carcinoma. AB - OBJECTIVE: The goal of this study was to assess the involvement of chromosome 1p deletion in ovarian papillary serous carcinoma (OPSC) via high-resolution physical mapping to detect a candidate tumor suppressor gene previously implicated in uterine papillary serous carcinoma (UPSC) tumorigenesis. METHODS: Previous studies have demonstrated a high rate of loss of heterozygosity (LOH) within a critical region of chromosome 1p in UPSC, suggesting the presence of a putative tumor suppressor gene important in the development of UPSC. To compare the genetic changes in OPSC with those in UPSC, seven microsatellite repeat polymorphisms were used to evaluate LOH in primary OPSC specimens. Allelic intensity was compared between normal and tumor DNA from microdissected, formalin fixed, paraffin-embedded specimens. In addition to the same seven 1p markers used for UPSC, three additional non-1p markers for chromosomes 1q, 11q, and 2p were tested to determine a baseline rate of LOH. RESULTS: Overall, 26.6% (8/30) of OPSC (vs 63.2% of UPSC) were characterized by loss at either of the two loci that define the critical region for UPSC. Rates of LOH at each of the 1p loci in the OPSC tumors ranged from 5.6 to 38.9%, which are compatible with background rates of loss for OPSC. LOH at non-1p loci was 29.2%. CONCLUSION: While a tumor suppressor gene on 1p appears to be an important genetic event in the development of UPSC, a similar rate and pattern of loss on 1p are not identified in OPSC. Thus, despite the striking clinical similarities between UPSC and OPSC, tumorigenesis in these carcinomas appears to occur via distinctly different pathways. PMID- 11426976 TI - The clinical significance of occult macroscopically positive retroperitoneal nodes in patients with epithelial ovarian cancer. AB - OBJECTIVE: The goal of this study was to determine the time during primary cytoreduction when retroperitoneal lymph nodes that are involved with macroscopic disease are recognized to be involved with tumor, the dimensions of intranodal disease present, and the possible clinical significance of macroscopically positive nodes that are recognized at different phases of the operation. METHODS: One hundred consecutive patients with stage IIIC and IV epithelial ovarian cancer underwent a retroperitoneal lymph node dissection during primary cytoreductive surgery. The phase of the operation in which nodes were recognized to be macroscopically involved with tumor was noted. Nodes were classified to be positive by palpation if recognized to be macroscopically involved by transperitoneal palpation, positive by inspection if recognized to be macroscopically involved by palpation after opening the retroperitoneal area, and positive by dissection if recognized to be macroscopically involved anytime after starting the actual process of lymph node dissection. The largest dimension of the intranodal disease in macroscopically positive nodes was measured. Log rank analysis determined whether nodal status or the time at which the nodes were recognized to be macroscopically positive influenced the probability of survival. RESULTS: Of the 100 patients, 66 had positive lymph nodes. Five were microscopically positive and 61 were macroscopically positive, of which 19 (31.1%) were positive by palpation, 16 (26.2%) were positive by inspection, 26 (42.6%) were positive by dissection. Of the 39 patients with negative and microscopically positive nodes 15 (38.5%) were clinically suspicious. Compared with patients with negative and microscopically positive lymph nodes, survival was not significantly different for patients who required excision of macroscopically positive nodal tissue. Survival was not influenced by the specific phase of surgery in which macroscopically positive nodes were recognized. CONCLUSIONS: A significant percentage of patients had retroperitoneal nodes recognized to be involved with macroscopic disease only after a lymph node dissection was in progress. The decision not to perform a lymph node dissection for optimally and completely cytoreduced patients may result in unrecognized macroscopic residual disease that is larger than what would otherwise be documented. PMID- 11426977 TI - Sigmoid perforation following radiation therapy in patients with cervical cancer. AB - OBJECTIVE: We describe the clinical presentation, evaluation, management, and outcome of patients experiencing sigmoid perforation following radiation therapy for cervical cancer. METHODS: A database consisting of over 5000 patients with stage IB-IIIB cervix cancer treated between 1963 and 1992 revealed 35 patients with sigmoid perforation. Twenty-seven were diagnosed and managed at one institution, and they form the study group. RESULTS: The median age at the time of perforation was 50 years, and the median follow-up care was 78 months (range 6 396). The median time from completion of radiotherapy to perforation was 13 months (range 3-98). The mean interval from the first documented complaint to the index admission was 90 days. Nine (33%) of 27 patients were treated with high dose radiation therapy. The most common complaint was abdominal pain in 25 (93%) patients, nausea occurred in 12 (44%) patients, weight loss in 12 (44%) patients, and vomiting in 10 (37%) patients. The pain was described as mild in 16 (73%) of 22 patients. Only 5 (18.5%) of 27 patients had physical signs of acute peritonitis, 8 (30%) of 27 patients had some form of tenderness, and 11 (41%) of 27 had a benign exam. A total of 20 (74%) patients had an abdominal radiograph, and 12 (44%) patients had a contrast enema for evaluation. Evidence of perforation was present in 5 (25%) of 20 plain abdominal radiographs and 1 (8%) of 12 contrast enemas. Following admission, 17 (63%) patients were observed initially with subsequent surgery after symptoms either failed to resolve or worsened. The median duration under observation was 4 days (range 1-23). Surgery was performed immediately in 8 patients (30%), and 2 (7%) were observed without operation. In these 2 patients, perforation was diagnosed postmortem. Seventeen (68%) of 25 patients had a localized abscess. Three of the patients who underwent immediate exploration and 7 who had surgery after a period of observation died postoperatively (10/25, 40%). Five (55%) of 9 patients in the group who received high-dose radiation therapy died because of sigmoid perforation. When the time frame of presentation was evaluated, we noted that 10 (50%) of 20 patients died between 1960 and 1979 and 1 (14%) of 7 died between 1980 and 1992. CONCLUSIONS: Sigmoid perforation following pelvic radiation for cervical cancer does not usually present with the typical signs of a ruptured viscus. A high degree of suspicion remains a priority in the care of radiated patients who present with abdominal pain given the atypical presentation of perforation in this group. PMID- 11426978 TI - Treatment of uterine papillary serous carcinoma with paclitaxel. AB - OBJECTIVE: The aim of this study was to determine the effectiveness and toxicity of monthly treatment with intravenous paclitaxel for women with advanced or recurrent uterine papillary serous carcinoma (UPSC). METHODS: Consenting women with histologically confirmed advanced (FIGO stage III or IV) or recurrent UPSC were treated on an Institutional Review Board approved protocol of a 24-h intravenous infusion of 200 mg/m(2) of paclitaxel every 3 weeks. Both measurable and nonmeasurable disease cases were enrolled. Treatment was continued until disease progression, patient intolerance, or (in women with nonmeasurable disease) completion of six courses. RESULTS: Twenty patients received from 1 to 11 cycles of therapy. Two women died of disease after 1 cycle of therapy and were not evaluable for response. Among 13 women with measurable tumor receiving 2 or more cycles of therapy, 4 had a complete clinical response and 6 had a partial response (objective response rate, 77%). The median time to progression was 7.3 months (range, 2-21 months). All 3 remaining patients with measurable disease had stable disease for a median of 6 months. The 5 patients without evaluable disease received 5 to 6 cycles of adjuvant paclitaxel. Three developed recurrence (range, 4-10 months; median, 7.2 months). Neutropenia was the major toxicity. Eleven of the 20 patients required G-CSF support, and 9 were hospitalized for neutropenic fever. One woman had reversible cardiac symptoms, which might have been related to paclitaxel treatment. At the time of analysis (mean follow-up, 23 months; range, 4.3-59.9 months), 13 women had died of disease, 4 were alive with disease, and 2 were disease free. All 3 disease-free patients had been treated for nonmeasurable advanced stage disease. CONCLUSION: Paclitaxel appears to have excellent activity in the treatment of advanced or recurrent UPSC, an uncommon but aggressive malignancy. Longer survival appears to be more common among women with small-volume disease. PMID- 11426979 TI - Vascular endothelial growth factor C gene expression is closely related to invasion phenotype in gynecological tumor cells. AB - OBJECTIVE: The correlation between the gene expression of various angiogenic factors and in vitro invasive activity in 16 human gynecological cancer cell lines was investigated. METHODS: Semiquantitative reverse transcription polymerase chain reaction analysis was performed to investigate the mRNA expression levels of vascular endothelial growth factors (VEGF-A, -B, -C, and D), basic fibroblast growth factor (bFGF), and matrix metalloproteinase (MMP)-2 with beta-actin coamplified as an internal standard. Tumor cell migration along a gradient of substratum-bound fibronectin and invasion into reconstituted basement membrane were evaluated by haptotactic migration and invasion assay. RESULTS: Expression of VEGF-A mRNA was detected in all 16 cell lines, whereas the relative expression levels of other VEGF family members and bFGF, differed markedly among the cell lines. There was a statistical correlation between VEGF-C gene expression and the number of cells that migrated and invaded (P < 0.01). However, expression of mRNAs of other angiogenic factors did not correlate with motility and invasive activity of the cells. Moreover, there was a close correlation between VEGF-C and MMP-2 gene expression levels (P < 0.05). CONCLUSION: Tumor cells that produce VEGF-C may have a higher invasive and metastatic potential because of their capacity to pass through tissue barriers. PMID- 11426980 TI - 10-Gy single-fraction pelvic irradiation for palliation and life prolongation in patients with cancer of the cervix and corpus uteri. AB - OBJECTIVE: The goal of this study was to evaluate the effects of single fractions of 10 Gy pelvic irradiation for palliation and life prolongation in patients with cancer of the uterine cervix or corpus. METHODS: A retrospective analysis was performed on 37 cervical cancer and 27 corpus cancer patients treated in the period 1988-1998. All patients had a life expectancy of less than 1 year. Due to stage of disease, age (median 82 years), or comorbidity they were considered unapt for surgery or conventional radiotherapy. Eleven patients with recurrence within irradiated field or early progression received one, 51 patients received two, and 2 patients received three fractions with a 4-week interval. RESULTS: Vaginal bleeding stopped in 90% and malodorous discharge in 39% of the patients. Among 46 patients with advanced disease treated for palliation, 10 (22%) showed complete tumor responses. Median time to progression was 6 months, and median survival, 9 months. Eighteen patients with early-stage disease and serious comorbidity were treated with life prolongation and symptom prevention as intention, and showed seven complete responses. Median survival was 13 months, and half the deaths were caused by intercurrent disease. Median hospital stay was 5 days and rehabilitation was easy. Fifty-six percent of the patients experienced no acute side effects and 33% had minor gastrointestinal problems. Three patients (6%) had serious late bowel complications, one with a fatal outcome, the symptoms appearing 9-10 months posttreatment. CONCLUSION: The 10-Gy single-fraction pelvic radiation regimen is an effective means of symptom palliation and is well tolerated. Tumor responses are obtained. The risk of late bowel complications is a concern for patients with a life expectancy greater than 9 months. PMID- 11426981 TI - Nitric oxide-dependent apoptosis in ovarian carcinoma cell lines. AB - OBJECTIVE: In a recent study, we found different profiles of inducible nitric oxide synthase (iNOS) gene expression in the ovarian carcinoma cell lines OVCAR 3, HOC-7, and 2774 following stimulation by proinflammatory cytokines. The present study was performed to determine whether nitric oxide (NO) synthesis correlates with programmed cell death in these cells. METHODS: NO-Dependent apoptosis was detected by DNA fragmentation analysis and fluorescence-activated cell sorter analysis. RESULTS: NO formation in response to interferon gamma (IFN gamma), interleukin-1beta (IL-1beta), and tumor necrosis factor alpha (TNF-alpha) was correlated with programmed cell death in the investigated cells. DNA fragmentation was most prominent in OVCAR-3 (34.17 +/- 1.81%), less pronounced in HOC-7 (12.86 +/- 0.45%), and undetectable in 2774 (4.54 +/- 0.40%) cells. The rate of apoptosis correlated with the amount of NO formation in cytokine-treated cells. Moreover, coincubation of OVCAR-3 and HOC-7 with the specific iNOS inhibitor aminoguanidine suppressed apoptosis induced by IFN-gamma, IL-1beta, and TNF-alpha. CONCLUSION: Our data indicate that in OVCAR-3 and HOC-7 cells, NO synthesis induced by IFN-gamma, IL-1beta, and TNF-alpha is correlated with the degree of apoptotic cell death. In clinical situations, this might in part explain the benefit of cytokine application in ovarian carcinoma patients (e.g., documented for IFN-gamma). PMID- 11426982 TI - Negative predictive value of human papillomavirus test following conization of the cervix uteri. AB - OBJECTIVE: The goal of this study was to determine/evaluate the negative predictive value of human papillomavirus (HPV) testing following conization of cervix uteri. METHODS: A prospective analysis was undertaken on 79 cone biopsies of women with high-grade lesions (cervical intraepithelial neoplasia (CIN) III). HPV testing was performed on cervical smears before and after conization. We correlated the margin status (defined as positive cone margin or endocervical curettage status) and positive conization HPV status with the residual disease in a hysterectomy specimen. A Digene II kit was used to perform HPV testing. HPV detection was done by Hybrid Capture assay. RESULTS: Of the 79 patients, 47(59.5%) had positive margins after conization. HPV testing was positive in 37 cases (78.7%) and negative in 10 cases (21.3%). Residual disease was found in 31 of 47 (66%) postconization hysterectomy specimens. No residual lesions were found in HPV-negative cases. Of the 32 cases with negative margins following conization, HPV testing was negative in 25 cases (78%) and was positive in 7 cases (22%). Among these 25 cases with negative HPV tests, no residual lesion was detected, and in 7 HPV-positive cases, only one residual lesion was found. CONCLUSION: HPV testing is potentially an effective tool in predicting residual dysplasia after conization and could potentially assist in the decision between hysterectomy and conservative follow-up in women with CIN III. PMID- 11426983 TI - Primary osteosarcoma of the uterine corpus: report of a case with immunohistochemical and ultrastructural study. AB - BACKGROUND: Primary uterine osteosarcoma is extremely rare, with only 15 cases reported in the literature. CASE: A 41-year-old woman presented with abnormal vaginal bleeding due to a large tumor arising from the uterine corpus. Histologically, the tumor showed the features of a malignant mesenchymal neoplasm with osteoid formation and lacked an epithelial component. Immunohistochemical and ultrastructural studies showed no evidence of epithelial differentiation. The tumor was excised and the patient received chemotherapy and radiation therapy postoperatively, but the tumor recurred locally at the 8-month follow-up. CONCLUSION: Osteosarcoma as a primary uterine tumor is exceedingly rare and should be distinguished from carcinosarcoma, which shows different macroscopic and histologic features. Prognosis of this neoplasm is very poor with an average life expectancy of 5 months. PMID- 11426984 TI - Large cell neuroendocrine cervical carcinoma: a report of two cases and review of the literature. AB - BACKGROUND: Large cell neuroendocrine cervical carcinoma is a rare malignancy. These tumors appear to mimic the aggressive behavior of small cell neuroendocrine tumors. Metastasis and recurrent disease are common. Due to the low incidence of these tumors, optimal therapy has not been delineated. CASES: Two patients presented with large cell neuroendocrine cervical carcinoma, stage IB1 and IIA, at our institution from 1997 to 1999. We describe the clinical course for these two patients and review the relevant literature for the management of large cell cervical carcinoma. CONCLUSION: Unlike squamous cell carcinoma, early-stage large cell neuroendocrine tumors of the cervix are aggressive. Disease recurrences are frequent and distant metastasis is common. Multimodal therapy should be considered at the time of initial diagnosis. PMID- 11426985 TI - Fatal recurrent ureteroarterial fistulas after exenteration for cervical cancer. AB - BACKGROUND: Ureteroarterial fistula (UAF) is a rare occurrence. It can be difficult to diagnose with a high mortality. We report a case of a recurrent UAF. CASE: A 38-year-old women diagnosed with cervical cancer had undergone pelvic exenteration for severe radiation-induced necrosis with a vesicovaginal and rectovaginal fistula after primary radiation therapy. Hemorrhage into the urinary tract necessitated surgical intervention and vascular repair with a femoral femoral bypass. Although these measures were effective, the patient died 6 months later following an acute hemorrhage into her conduit. Arteriogram revealed a second UAF. CONCLUSION: When urinary tract bleeding occurs in patients previously diagnosed with a gynecologic malignancy and treated with radiation therapy and extensive surgery with urinary diversion, UAF should be considered in the differential diagnoses. PMID- 11426986 TI - Bladder carcinoma associated with cyclophosphamide therapy for ovarian cancer occurring with a latency of 20 years. AB - BACKGROUND: Cyclophosphamide-induced urothelium cancer of the bladder is a well known entity. The risk for inducing such cancer grows with duration and dosage of cyclophosphamide therapy. The lag time between termination of treatment and development of urothelial cancer has been observed to be between 9 months and 11 years. Single cases have been reported 14, 16, 17, and 21 years after cyclophosphamide treatment. CASE: We report a case of a bladder cancer occurring after a lag time of 20 years after oral therapy with cyclophosphamide for ovarian cancer. The bladder cancer was detected due to gross hematuria. CONCLUSION: It is of great importance for gynecologists to continue to care for patients who have received long-term cyclophosphamide treatment, even if the treatment was completed decades ago. One possible early symptom of cyclophosphamide-induced bladder cancer is painless hematuria. This can easily be used to detect bladder cancer in women at risk, even after a very long latency period. PMID- 11426987 TI - Transitional cell carcinoma of the bladder mimicking recurrent paget's disease of the vulva: report of two cases, with one occurring in a myocutaneous flap. AB - BACKGROUND: Transitional cell carcinoma of the bladder may spread superficially along and beyond the urogenital epithelium, mimicking vulvar Paget's disease. CASES: These two cases illustrate unusual aspects of transitional cell carcinoma of the bladder and vulvar Paget's disease. Both patients had a history of breast cancer and previously had multiple operations for recurrent vulvar Paget's disease; one patient had a radical vulvectomy with transverse rectus abdominal muscle flap reconstruction. Both had a history of recurrent transitional cell carcinoma of the bladder. Both presented with recalcitrant transitional cell carcinoma of the bladder and clinically recurrent vulvar Paget's disease. Pathologic evaluation, however, revealed pagetoid spread of carcinoma in situ (CIS) throughout the urothelium, with an invasive component in the cervix and extension of the CIS into the rectum in one patient. CONCLUSION: If the history of the patient includes transitional cell carcinoma of the bladder and vulvar Paget's disease, histologic evaluation is needed for accurate diagnosis and proper treatment. PMID- 11426988 TI - Is the invasion depth in millimeters valid to determine the prognosis of early invasive cervical adenocarcinoma? A case of recurrent FIGO stage IA1 cervical adenocarcinoma. AB - BACKGROUND: Recurrence of FIGO stage IA1 cervical adenocarcinoma is extremely rare. We herein report a patient with early invasive cervical adenocarcinoma who developed a recurrence in the vaginal stump. CASE: A 52-year-old female complained of contact bleeding. Biopsy of the uterine cervix verified cervical adenocarcinoma, and the patient underwent Okabayashi hysterectomy with pelvic lymphadnectomy and bilateral adnectomy. Histopathologic examination of the uterus revealed an invasive cancer 3 mm in depth. Neither lymph node metastasis nor lymph-vascular space invasion was observed. However, the depth of her normal cervical gland area was 2 mm only, and the cancer invasion involved an area which was deeper than the normal cervical gland area. The vaginal stump recurrence developed 4 years after surgery. CONCLUSION: The depth of invasion with reference to that of normal cervical glands may become a possible prognostic factor for early invasive cervical adenocarcinoma. PMID- 11426989 TI - Cutaneous metastasis of papillary serous uterine cancer. AB - BACKGROUND: Cutaneous metastases from uterine cancer are uncommon. They usually indicate the aggressive nature of the underlying disease. CASE: A 65-year-old patient presented with stage 3 papillary serous carcinoma of the uterus. Eight months after surgery and pelvic radiation therapy, this patient presented with cutaneous metastases. A single cobalt field was used on three occasions over the next 9 months to provide symptomatic control of the abdominal and vulvar cutaneous metastases. CONCLUSIONS: This is the first case report of cutaneous metastases from papillary serous cancer of the uterus. Cutaneous metastases were the first indication of unsuspected occult disease. Palliative local radiation has a useful role in controlling symptoms like pruritus. PMID- 11426990 TI - Metastatic serous borderline ovarian tumor in an internal mammary lymph node: a case report and review of the literature. AB - Background. Reports of supradiaphragmatic involvement of lymph nodes by serous borderline ovarian tumors are rare. Case. We describe the finding of metastatic tumor involving an internal mammary lymph node in a 74-year-old patient with serous borderline ovarian tumor. The positive lymph node was found incidentally during cardiac surgery 7 years after excision of the patient's serous borderline ovarian tumor. The incidence and significance of pelvic, paraaortic, and supradiaphragmatic lymph node involvement among women with borderline ovarian tumors is discussed. Conclusion. Supradiaphragmatic lymph node involvement can occur among women with serous borderline ovarian tumors. Borderline ovarian tumors should be included in the differential diagnosis of metastatic adenocarcinoma involving the supradiaphragmatic lymph nodes. PMID- 11426991 TI - Endometrial carcinoma and an unusual presentation of bone metastasis: a case report. AB - BACKGROUND: Although metastasis to bone is common in solid tumors, it seldom occurs in endometrial carcinoma. It is usually seen together with abdominopelvic recurrences and/or other organ metastases. Furthermore, bone metastases involving only the lower limbs are seen extremely rare. CASE: 67-year-old woman was referred for a vaginal recurrence from endometrial carcinoma. Pelvic irradiation and intracavitary brachytherapy failed to control the disease. Subsequently, she was presented with swelling of her left foot resembling an abscess, and bone scan revealed an increased uptake in both lower extremities below the knees. Furthermore, biopsy showed metastatic disease. She died 2 months after palliative irradiation with progressive disease. CONCLUSION: It should be remembered that bone metastasis at unusual sites might be seen in endometrial carcinoma. PMID- 11426992 TI - Primitive Model Electrophoresis. AB - The electrophoresis of a spherical macroion is calculated with the new primitive model electrophoresis (PME) theory, which incorporates the ionic size. The results are compared with the classical theory of Wiersema, O'Brien, and White. The PME mobility, as a function of the macroion's surface charge (sigma) or zeta potential (zeta), is found to depend on the ionic valence, radius and concentration, and macroion hydrodynamical radius (A); i.e., it is not universal with kappaA, as predicted by the classical theory. The mobility is very nonlinear as a function of zeta. This behavior is related to the nonlinear dependence of zeta, as a function of sigma, when ionic size is included in the theory. In the classical theory zeta is a monotonic function of sigma. Important quantitative and/or qualitative differences between PME and the classical theory are found. However, in the limit of zero ionic diameter, and/or low salt concentration, and low macroion charge, the new theory reduces to the classical theory. The agreement of the PME with experimental data is very good. In particular, the prediction of reversed mobility is corroborated by recent experimental data. Copyright 2001 Academic Press. PMID- 11426993 TI - A Thermodynamically Consistent Langmuir Model for Mixed Gas Adsorption. AB - A multiregion, extended Langmuir model (MREL) is proposed for predicting multicomponent adsorption equilibria. For a binary mixture, it is assumed that two regions or sites exist on the surfaces. One region or site adsorbs both species from the mixture whereas the other region or site is excluded from one species due to size exclusion or energetic exclusion (or lack of competition). On the region where both species are adsorbed, the saturated adsorption capacities are the same, and consequently the extended Langmuir model (EL) applied to this region attains thermodynamic consistency. The model is also extended to multicomponent mixtures. The total adsorbed amount of a component is the sum of that adsorbed on all regions/sites. Only the two parameters of the Langmuir isotherm for each pure component are used. The MREL model is computationally as simple as the EL model. Eight binary and one ternary systems, including some highly nonideal ones, have been tested, and significant improvements are seen with the MREL over the EL. In particular, the MREL is capable of predicting the azeotropic behavior of highly nonideal systems. Copyright 2001 Academic Press. PMID- 11426994 TI - Sorption of Nickel on Carbonate Fluoroapatites. AB - The retention properties of a synthetic carbonate fluoroapatite and a natural francolite are compared in the present work from an investigation of the sorption of (63)Ni at tracer scale amounts onto these solids. Two different surface complexation models were successively used to fit the experimental adsorption isotherms obtained under various experimental conditions: the nonelectrostatic model and the constant capacitance model. The results are essentially described by two main equilibria involving one proton in acidic media and three protons in basic media. The corresponding thermodynamic constants are in agreement for both models. Modeling gives also close values for both solids, despite their distinct solubility and surface acidity. Copyright 2001 Academic Press. PMID- 11426995 TI - On the Temperature Dependence of Intrinsic Surface Protonation Equilibrium Constants: An Extension of the Revised MUSIC Model. AB - The revised multisite complexation (MUSIC) model of T. Hiemstra et al. (J. Colloid Interface Sci. 184, 680 (1996)) is the most thoroughly developed approach to date that explicitly considers the protonation behavior of the various types of hydroxyl groups known to exist on mineral surfaces. We have extended their revised MUSIC model to temperatures other than 25 degrees C to help rationalize the adsorption data we have been collecting for various metal oxides, including rutile and magnetite to 300 degrees C. Temperature-corrected MUSIC model A constants were calculated using a consistent set of solution protonation reactions with equilibrium constants that are reasonably well known as a function of temperature. A critical component of this approach was to incorporate an empirical correction factor that accounts for the observed decrease in cation hydration number with increasing temperature. This extension of the revised MUSIC model matches our experimentally determined pH of zero net proton charge pH values (pH(znpc)) for rutile to within 0.05 pH units between 25 and 250 degrees C and for magnetite within 0.2 pH units between 50 and 290 degrees C. Moreover, combining the MUSIC-model-derived surface protonation constants with the basic Stern description of electrical double-layer structure results in a good fit to our experimental rutile surface protonation data for all conditions investigated (25 to 250 degrees C, and 0.03 to 1.0 m NaCl or tetramethylammonium chloride media). Consequently, this approach should be useful in other instances where it is necessary to describe and/or predict the adsorption behavior of metal oxide surfaces over a wide temperature range. Copyright 2001 Academic Press. PMID- 11426996 TI - Heteroflocculation of Sulfate Polystyrene Latex and Anticarsia gemmatalis Nucleopolyhedrovirus as a Model System for Studying Sunlight Protection. AB - Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) is a baculovirus widely used as a pest control agent. Data on field persistence of this microbial insecticide reveal that sunlight is the most destructive of the environmental factors. The introduction of a physical barrier to protect the baculovirus from sunlight has been proposed. In the present work we report on the coating of a baculovirus surface with a uniform layer of smaller particles. The coating was carried out at pH 3, a condition where the baculovirus surface becomes positively charged and can be covered by sulfate polystyrene latex particles. The heteroflocculation was assessed by isotherm measurements and scanning electron microscopy. High-affinity isotherms were obtained for the two sizes of latex used, and the number of latex particles bound per baculovirus was not affected when pH was changed back to neutral range (50.6-0.8. The compared selectivities of Ca and Mg for this smectite are nearly equal. As far as the exchanger is concerned, the Na-Ca and Na-Mg exchange is nearly ideal, in the raoultian sense. Thermodynamic exchange constants and the corresponding Gibbs free energies of reaction have been computed from (i)(j)K(V) coefficients. This allow us to quantify the exchanger's affinity sequence, which appears to be Na(+)90% for BCG and >65% for M. leprae). The early activation of TKs during mycobacteria-host cell interaction was confirmed by immunoblot analysis, demonstrating that in several host cell proteins mycobacteria stimulated tyrosine phosphorylation. The use of the drugs wortmannin and bisindolylmaleimide I which, respectively, inhibit phosphatidylinositide 3-kinase (PI 3-kinase) and protein kinase C (PKC), produced lower but consistent results within a 35--60% association inhibition range for both bacteria. Dose response curves with these inhibitors were obtained. Similar results were obtained when primary human monocytes were used as host cells, strongly suggesting that TK, PKC and PI 3 kinase signals are activated during the interaction of human monocytes with both pathogenic and attenuated species of mycobacteria. PMID- 11427036 TI - The effect of revascularisation of an ischaemic lower limb on leukotriene E4 production. AB - It has previously been shown that leukotriene E4 production is increased both in acute and chronic lower limb ischaemia. The aim of this study was to measure the effect of revascularisation on leuktriene E4 excretion in chronic lower limb ischaemia. Revascularisation did not affect significantly on leukotriene E4 excretion (preop. 34.9+/-7.1 pg/mg creatinine, postop. 24.5+/-4.7 pg/mg creatinine, n=10, P<0.238). We suggest that the enhanced leukotriene E4 production continues after revascularisation which may have a therapeutical implication. PMID- 11427037 TI - Effect of low-dose aspirin in combination with stable fish oil on whole blood production of eicosanoids. AB - The effect of a combination of aspirin and fish oil on eicosanoids was studied. Four subjects were given 37.5 mg aspirin orally, and 6 weeks later they received a natural, stable fish oil daily for 1 week before taking the same single dose of aspirin. Blood samples for determination of whole blood production of eicosanoids were taken before and after each experimental period. Serum thromboxane A(2)was decreased by 40% (P<0.05) after aspirin alone, but by 62% (P<0.01) after fish oil + aspirin. Serum prostacyclin (measured as 6-keto PGF(1a)) was decreased by aspirin in both cases. The sum of 6-keto PGF(1a)and its equipotent fish oil derived analogue Delta(17)-6-keto PGF(1a)was reduced after aspirin intake (55%, NS), but after fish oil + aspirin the reduction was smaller (33%, NS). Leukotriene B(4)was increased by 19% (P<0.05) after aspirin, and decreased by 69% (P<0.05) after fish oil + aspirin. A combination of stable fish oil and aspirin thus improves the eicosanoid pattern more than aspirin alone. PMID- 11427038 TI - Do human platelets express COX-2? AB - The rate-limiting enzyme in prostaglandin (PG)- and thromboxane (TX)-synthesis is known as cyclooxygenase (COX). The COX-enzyme family consists of the classical COX-1 and the inducible COX-2-enzyme. To investigate whether platelets contain COX-2, we measured thiobarbituric acid reactive substances (TBARS) after either blocking COX-1 or COX-2 or adding compounds known to affect COX-expression. To stimulate platelets' different reagents such as collagen, thrombin and arachidonic acid (AA) were used. The inhibitors used in this study were acetylsalicylic acid (ASA), indomethacin and NS-398. Using the western-blot technique, we failed to detect COX-2 in platelets while COX-1 was detectable. We were not able to discover COX-2 in platelets using the methods we applied. As the amount of COX-2 in platelets might be below the detection limit of the methods used, the biological relevance COX-2 in platelets, if even existing at low amounts, remains to be established. PMID- 11427039 TI - Increase of isoprostane 8-epi-PGF(2alpha)after restarting smoking. AB - Isoprostanes are known as reliable markers of in vivo oxidation injury. Cigarette smoking has been shown to be associated with a significant increase in 8-epi PGF(2alpha), a major member of this family of compounds. Quitting smoking reduces 8-epi-PGF(2alpha) values to normal within a couple of weeks only. In this follow up we checked the 8-epi-PGF(2alpha), values in plasma, serum and urine in 28 people who restarted smoking after a quitting attempt of various duration. 8-epi PGF(2alpha)shows a certain increase after restarting smoking reaching a maximum after already 1 week. Continuation of smoking does not significantly further increase 8-epi-PGF(2alpha). These data indicate a fast response of restarting as on quitting smoking on in vivo oxidation injury. The oxidation injury reflected by 8-epi-PGF(2alpha)may be a key pathogenetic mechanism in smoking-induced vascular injury. PMID- 11427040 TI - Involvement of inducible isoforms of COX and NOS in streptozotocin-pancreatic damage in the rat: interactions between nitridergic and prostanoid pathway. AB - Streptozotocin-induced pancreatic damage involves nitric oxide (NO) and prostaglandins (PGs) overproduction. In this work we aim to evaluate a putative relationship between the elevated NO levels and the altered prostanoid production in pancreatic tissue from streptozotocin-diabetic rats. Total NOS activity and nitrate/nitrite pancreatic levels in tissues from diabetic rats are decreased when the cyclooxygenase (COX) inhibitor indomethacin (INDO) is added to the incubating medium, while the addition of PGE(2)increases nitrate/nitrite production and NOS levels. INDO and PGE(2)selectively affect Ca(2+)-dependent NOS (iNOS) activity in diabetic tissues, and they have not been able to modify nitrate/nitrite levels, iNOS or Ca(2+)-dependent (cNOS) in control tissues. When the NOS inhibitor L-NMMA was present in the incubating medium, control pancreatic [(14)C]-Arachidonic Acid ([(14)C]-AA) conversion to 6-keto PGF(1 alpha)and to TXB(2)was lower, and PGF(2 alpha), PGE(2)and TXB(2)production from diabetic tissues diminished. The NO donors, spermine nonoate (SN) and SIN-1, enhanced TXB(2)levels in control tissues, while PGF(2 alpha), PGE(2)and TXB(2)levels from diabetic tissues were increased. PGE(2)production from control and diabetic tissues was assessed in the presence of the NO donor SN plus INDO or NS398, a specific PG synthase 2 inhibitor. When SN combined with INDO or NS398 was added, the increment of PGE(2)production was abolished by both inhibitors in equal amounts, indicating that the activating effect of nitric oxide is exerted on the inducible isoform of cyclooxygenase. In the diabetic rat, prostaglandins and NO seem to stimulate the generation of each other, suggesting a lack of regulatory mechanisms that control the levels of vasoactive substances in acute phase of beta-cell destruction. PMID- 11427041 TI - Urinary PGE(2) concentrations measured by a new EIA method in infants with urinary tract infections or renal malformations. AB - In this work PGE(2)concentrations were measured by a new EIA method in the urine of infants (mean age: 9.35+/-4.24 months) with recurrent urinary tract infections or renal malformations. Compared to healthy subjects, PGE(2)excretion rates resulted significantly higher in both pathological groups, in particular in subjects with obstructive uropathies (29.55+/-8.12 vs 18.37+/-4.64 pg/ml). We did not find any age- or pH-dependent difference in urinary excretion of PGE(2); none of the examined indices of renal function showed any significant relationship to PGE(2). These results suggest that this parameter, measured non-invasively in the urine, could help in the differential diagnosis between obstructive vs non obstructive dilatation and in monitoring renal function in presence of recurrent UTI episodes. PMID- 11427042 TI - Plant sterols regulate rat vascular smooth muscle cell growth and prostacyclin release in culture. AB - Cardiovascular disease (CVD) is the leading cause of death in the USA and other industrialized countries. A large number of epidemiological studies have established a direct correlation between diet and the development and progression of atherosclerosis. Several studies have shown the incidence of CVD to be lower in populations consuming a predominantly plant-based diet, as compared to meat based diets. Besides being low in fat and cholesterol, vegetarian and Asian diets contain a large variety of phytochemicals, which may function in the body. For example, phytosterols (PS) are plant sterols that interfere with the absorption of cholesterol from the intestine when present in adequate amounts. Although PS may also function at a cellular level in the body, there are few studies examining the action of PS on cells involved in atherosclerosis. The purpose of this study was to examine the effect of dietary PS on vascular smooth muscle cell (VSMC) growth and function, since VSMC play a central role in the development of atherosclerosis. VSMC were treated with 16 microM cholesterol, 25 hydroxycholesterol, campesterol and beta-sitosterol (SIT) using an ethanol as a vehicle. Cell growth was determined by cell counting and cell proliferation by DNA synthesis, which was measured by [(3)H]-thymidine incorporation. Cholesterol supplementation had no effect on cell growth and proliferation. 25 Hydroxycholesterol decreased cell growth by 68% and DNA synthesis by 99%. SIT was found to inhibit VSMC growth more effectively than campesterol. Of the two PS, campesterol decreased cell growth by 16% and SIT decreased cell growth by 30%. DNA synthesis was decreased 25% by SIT supplementation but was not influenced by campesterol or cholesterol supplementation. Cholesterol, campesterol and SIT were not cytotoxic to VSMC and did not significantly alter cell viability. 25 Hydroxycholesterol, however, was cytotoxic and decreased cell viability by 45% as determined by lactate dehydrogenase release and a trypan blue dye exclusion test. De novo cholesterol synthesis was decreased 28% by campesterol, 49% by SIT and 23% by cholesterol. Beta-sitosterol exhibited a greater effect on cholesterol synthesis than campesterol or cholesterol supplementation. Measurement of cell sterol content demonstrated incorporation of PS into VSMC at the expense of cholesterol. Campesterol decreased VSMC cholesterol content by 36%, representing 40% of the total sterol content following treatment. Beta-sitosterol decreased VSMC cholesterol by 41% following supplementation and represented 49% of the total sterol amount. Cholesterol treatment did not alter the cholesterol content of the cells. Prostacyclin production was significantly altered by PS treatment. Basal prostacyclin release was increased 43% by campesterol and 81% by SIT. A23187 stimulated prostacyclin release was increased 25% by campesterol and 54% by SIT. SIT supplementation induced a greater effect on prostacyclin release from VSMC than cholesterol or campesterol supplementation. The in vitro results presented here suggest that dietary PS, especially SIT, may offer protection from the VSMC hyperproliferation found in atherosclerosis. Further in vivo research is needed to support these observations. PMID- 11427043 TI - Rational approaches to the design of cationic gemini surfactants for gene delivery. AB - We report a new class of amphiphilic gemini surfactants as vehicles for gene delivery into cells, and the beginnings of a systematic structure-activity study. Preliminary results suggest that combining gemini surfactants with dioleoylphosphatidylethanolamine (DOPE) should allow the preparation of liposomes of various sizes and lipid compositions. Control of such colloidal changes could be as significant as the changes in the molecular composition of the gemini surfactants in delivering optimum gene expression in animal models. PMID- 11427044 TI - Facilitated phosphatidylcholine flip-flop across erythrocyte membranes using low molecular weight synthetic translocases. AB - The transmembrane distribution of phospholipids plays an important regulatory role in human erythrocytes. Membrane-bound translocase enzymes maintain an asymmetric phospholipid distribution across the membrane monolayers by promoting transmembrane diffusion or flip-flop. Mechanistic understanding of the flip-flop process is weak at the molecular level. Recently, we discovered that amide and sulfonamide derivatives of tris(aminoethyl)amine facilitate phospholipid flip flop across vesicle membranes; that is, they act as low molecular weight, synthetic translocases. In this report, NMR evidence is provided that suggests that the synthetic translocases work by forming a hydrogen-bonded complex with the phosphocholine headgroup which decreases headgroup polarity and promotes diffusion across the lipophilic interior of the membrane. Also cell morphology and fluorescence probe methods are used to show that these synthetic translocases facilitate phosphatidylcholine flip-flop across erythrocyte membranes. Addition of a small amount of dilauroylphosphatidylcholine to erythrocytes produces echinocyte morphology which takes days to revert back to the original discocyte shape. The rate of return is significantly accelerated by the presence of the synthetic translocases. The synthetic translocases facilitate inward translocation (flip) of the fluorescent phosphatidylcholine probe, 1-palmitoyl-2 (N-[7-nitrobenz-2-oxa-1,3-diazol-4-yl]aminohexanoyl)-sn-glycero-3-phosphocholine (PC-NBD). PMID- 11427045 TI - Direct measurement indicates a slow cis/trans isomerization at the secondary amide peptide bond of glycylglycine. AB - Spectral differences between the cis and the trans isomer of a secondary amide peptide bond were used to follow the time course of the cis/trans isomerization of Gly-Gly, Gly-Ala, Ala-Gly, and Ala-Ala dipeptides in the UV/vis region at 220 nm. Isomerization rates and Eyring activation energies were calculated from pH- and LiCl-mediated solvent jump experiments. Rate constants were found to be in a narrow range of 0.29 to 0.64 s(-)(1) for the zwitterionic dipeptides at 25 degrees C. The isomerization rate is about 2-fold higher for the monoionic forms of Gly-Gly. The zwitterionic Gly-Gly has an activation enthalpy DeltaH() of 71.6 +/- 4.9 kJ mol(-)(1) that is in the range of the rotational barriers of aromatic side chain dipeptides that have been measured by (1)H NMR magnetization transfer experiments. Late stages of protein backbone rearrangements often involve crossing the energy barrier for rotational isomerization of imidic peptide bonds. Our findings are consistent with the idea that a wide range of secondary amide peptide bonds are also able to induce slow rate-limiting steps in protein restructuring. PMID- 11427046 TI - The role of cation-pi interactions in biomolecular association. Design of peptides favoring interactions between cationic and aromatic amino acid side chains. AB - Cation-pi interactions between amino acid side chains are increasingly being recognized as important structural and functional features of proteins and other biomolecules. Although these interactions have been found in static protein structures, they have not yet been detected in dynamic biomolecular systems. We determined, by (1)H NMR spectroscopic titrations, the energies of cation-pi interactions of the amino acid derivative AcLysOMe (1) with AcPheOEt (2) and with AcTyrOEt (3) in aqueous and three organic solvents. The interaction energy is substantial; it ranges from -2.1 to -3.4 kcal/mol and depends only slightly on the dielectric constant of the solvent. To assess the effects of auxiliary interactions and structural preorganization on formation of cation-pi interactions, we studied these interactions in the association of pentapeptides. Upon binding of the positively-charged peptide AcLysLysLysLysLysNH(2) (5) to the negatively-charged partner AcAspAspXAspAspNH(2) (6), in which X is Leu (6a), Tyr (6b), and Phe (6c), multiple interactions occur. Association of the two pentapeptides is dynamic. Free peptides and their complex are in fast exchange on the NMR time-scale, and 2D (1)H ROESY spectra of the complex of the two pentapeptides do not show intermolecular ROESY peaks. Perturbations of the chemical shifts indicated that the aromatic groups in peptides 6b and 6c were affected by the association with 5. The association constants K(A) for 5 with 6a and with 6b are nearly equal, (4.0 +/- 0.7) x 10(3) and (5.0 +/- 1.0) x 10(3) M( )(1), respectively, while K(A) for 5 with 6c is larger, (8.3 +/- 1.3) x 10(3) M( )(1). Molecular-dynamics (MD) simulations of the pentapeptide pairs confirmed that their association is dynamic and showed that cation-pi contacts between the two peptides are stereochemically possible. A transient complex between 5 and 6 with a prominent cation-pi interaction, obtained from MD simulations, was used as a template to design cyclic peptides C(X) featuring persistent cation-pi interactions. The cyclic peptide C(X) had a sequence in which X is Tyr, Phe, and Leu. The first two peptides do, but the third does not, contain the aromatic residue capable of interacting with a cationic Lys residue. This covalent construct offered conformational stability over the noncovalent complexes and allowed thorough studies by 2D NMR spectroscopy. Multiple conformations of the cyclic peptides C(Tyr) and C(Phe) are in slow exchange on the NMR time-scale. In one of these conformations, cation-pi interaction between Lys3 and Tyr9/Phe9 is clearly evident. Multiple NOEs between the side chains of residues 3 and 9 are observed; chemical-shift changes are consistent with the placement of the side chain of Lys3 over the aromatic ring. In contrast, the cyclic peptide C(Leu) showed no evidence for close approach of the side chains of Lys3 and Leu9. The cation-pi interaction persists in both DMSO and aqueous solvents. When the disulfide bond in the cyclic peptide C(Phe) was removed, the cation-pi interaction in the acyclic peptide AC(Phe) remained. To test the reliability of the pK(a) criterion for the existence of cation-pi interactions, we determined residue-specific pK(a) values of all four Lys side chains in all three cyclic peptides C(X). While NOE cross-peaks and perturbations of the chemical shifts clearly show the existence of the cation-pi interaction, pK(a) values of Lys3 in C(Tyr) and in C(Phe) differ only marginally from those values of other lysines in these dynamic peptides. Our experimental results with dynamic peptide systems highlight the role of cation-pi interactions in both intermolecular recognition at the protein-protein interface and intramolecular processes such as protein folding. PMID- 11427047 TI - Calculation of relative binding free energy differences for fructose 1,6 bisphosphatase inhibitors using the thermodynamic cycle perturbation approach. AB - An iterative, computer-assisted, drug design strategy that combines molecular design, molecular mechanics, molecular dynamics (MD), and free energy perturbation (FEP) calculations with compound synthesis, biochemical testing of inhibitors, and crystallographic structure determination of protein-inhibitor complexes was successfully used to predict the rank order of a series of nucleoside monophosphate analogues as fructose 1,6-bisphosphatase (FBPase) inhibitors. The X-ray structure of FBPase complexed with 5-aminoimidazole-4 carboxamide-1-beta-D-ribofuranosyl 5'-monophosphate (ZMP) provided structural information used for subsequent analogue design and free energy calculations. The FEP protocol was validated by calculating the free energy differences for the mutation of ZMP (1) to AMP (2). The calculated results showed a net gain of 1.7 kcal/mol, which agreed with the experimental result of 1.3 kcal/mol. FEP calculations were performed for 18 other AMP analogues. Inhibition constants were determined for over half of these analogues, usually after completion of the calculation, and were consistent with the predictions. Solvation free energy differences between AMP and various AMP analogues proved to be an important factor in binding free energies, suggesting that increased desolvation costs associated with the addition of polar groups to an inhibitor must be overcome by stronger ligand-protein interactions if the structural modification is to enhance inhibitor potency. The results indicate that FEP calculations predict relative binding affinities with high accuracy and provide valuable insight into the factors that influence inhibitor binding and therefore should greatly aid efforts to optimize initial lead compounds and reduce the time required for the discovery of new drug candidates. PMID- 11427048 TI - Computing handedness: quantized and superposed switch and dynamic memory of helical polysilylene. AB - Two new conjugating helical polymers comprising a rodlike silicon backbone and enantiopure chiral pendants, poly[(R)-3,7-dimethyloctyl-(S)-3 methylpentylsilylene] (PS-1) and its diastereomeric poly[(S)-3,7-dimethyloctyl (S)-3-methylpentylsilylene] (PS-2), were prepared. Molecular mechanics calculations of PS-1 and PS-2 model oligomers indicated a double well potential energy curve corresponding to almost enantiomeric helices with dihedral angles of 150-160 degrees (P-motif, global minimum) and 200-210 degrees (M-motif), regardless of their tacticity. Experimentally, it was found that PS-1 in dilute isooctane revealed switchable ambidextrous helicity on application of a thermal energy bias. Although PS-1 featured three distinct switching regions, viz. "region 1, between -80 and -10 degrees C", "region 2, between -10 and +10 degrees C", and "region 3, between +10 degrees C and +80 degrees C", the switching properties were interpreted as the result of superposed P- and M-helicities, undergoing dynamic pseudo-racemization or oscillation. Oscillating helicity in region 2 was roughly estimated to be about 13 cm(-)(1). The superposed helicity in region 2 was critical since it afforded molecular recognition ability with a dynamic memory function that was highly susceptible to solvent molecular topology and volume fraction. This could lead to potential as a molecular information processor to serve as a gauge of chemical properties. On the other hand, PS-2 could not switch its preferential screw-sense in the range of -80 to +80 degrees C. This may be related to greater differences the potential energy curve between P- and M-motifs. PMID- 11427049 TI - An enthalpic component in cooperativity: the relationship between enthalpy, entropy, and noncovalent structure in weak associations. AB - Attempts to quantify binding interactions of noncovalent complexes in aqueous solution have been stymied by complications arising from enthalpy-entropy compensation and cooperativity. We have extended work detailing the relationship between noncovalent structure and free energy of binding to include the roles of enthalpy and entropy of association. On the basis of van't Hoff measurements of the dimerization of vancomycin type antibiotics, we demonstrate that positive cooperativity manifests itself in a more favorable enthalpy of association and a partially compensating less favorable entropy of association. Finally, we extend these results to rationalize thermodynamic observations in unrelated systems. PMID- 11427050 TI - Synthesis of nitrogen analogues of salacinol and their evaluation as glycosidase inhibitors. AB - The syntheses of two nitrogen analogues (11 and 12) of the naturally occurring sulfonium ion, salacinol (7) are described. The latter compound is one of the active principles in the aqueous extracts of Salacia reticulata that are traditionally used in Sri Lanka and India for the treatment of diabetes. The synthetic strategy relies on the nucleophilic attack of a 1,4-dideoxy-1,4-imino-D or L-arabinitol at the least hindered carbon of 2,4-O-benzylidene D- or L erythritol-1,3-cyclic sulfate. The nitrogen analogues bear a permanent positive charge and serve as mimics of the sulfonium ion. We reasoned that these ammonium derivatives should function in a manner similar to that of known glycosidase inhibitors of the alkaloid class such as castanospermine (4) and deoxynojirimycin (5). Enzyme inhibition assays indicate that salacinol (7) is a weak (K(i) = 1.7 mM) inhibitor of glucoamylase, whereas compounds 11 and 12 inhibit glucoamylase with K(i) values in the range approximately 10-fold higher. The nitrogen analogues 11 and 12 showed no significant inhibitory effect of either barley alpha-amylase (AMY1) or porcine pancreatic alpha-amylase (PPA) at concentrations of 5 mM. In contrast, salacinol (7) inhibited AMY1 and PPA in the micromolar range, with K(i) values of 15 +/- 1 and 10 +/- 2 microM, respectively. PMID- 11427051 TI - The elusive 16-electron Cp*M(NO)Me(2)(M = Mo, W) complexes and their spontaneous conversions to Cp*M(NMe)(O)Me isomers. AB - Treatment of [Cp*Mo(NO)Cl(mu-Cl)](2) with magnesium (Me(2)Mg.dioxane, MeMgCl) or aluminum (Me(3)Al) methylating reagents affords the known compound [Cp*Mo(NO)Me(mu-Cl)](2) (1). Similar treatment of the dichloro precursor with MeLi in ethereal solvents generates an equimolar mixture of 1 and the trimethyl "ate" complex, Cp*MoMe(3)(NO-Li(OEt(2)(n)), (2-Et(2)O). Reaction of 2-Et(2)O with a source of [Me](+) forms Cp*MoMe(3)(=N-OMe)(3), a rare terminal alkoxylimido complex. Metathesis of the chloro ligands of [Cp*Mo(NO)Cl(mu-Cl)](2) by MeLi in toluene at low temperatures produces the target dimethyl complex, Cp*Mo(NO)Me(2) (4), in 75% isolated yield. In solution, 4 is predominantly a monomeric species, whereas in the solid state it adopts a dimeric or oligomeric structure containing isonitrosyl bridges as indicated by IR and (15)N/(13)C NMR spectroscopies. Hydrolysis of 4 affords meso- and rac-[Cp*Mo(NO)Me](2)(mu-O) (5), and the reactions of 4 with a range of Lewis bases, L, to form the 18e adducts Cp*Mo(NO)(L)Me(2) (e.g., Cp*Mo(NO)(PMe(3))Me(2) (7)), have established it to be the most electrophilic complex of its family. Acidolysis of the methyl groups of 4 is also facile. Most notably, 4 is thermally unstable in solution and undergoes isomerization via nitrosyl N-O bond cleavage to its oxo(imido) form, Cp*Mo(NMe)(O)Me (11), which is isolable from the final reaction mixture as the mu oxo-bridged adduct formed by 4 and 11, i.e., Cp*Mo(NO)Me(2)(mu-O)Cp*Mo(NMe)Me (4 <-- 11). The rate of this isomerization is significantly faster for the tungsten dimethyl complex; hence, Cp*W(NO)Me(2) (12) is not isolable free of a supporting donor interaction and can only be isolated as Cp*W(NO)Me(2)(mu-O)Cp*W(NMe)Me (12 <-- 13) or Cp*W(NO)Me(2)(PMe(3)) (14) adducts. PMID- 11427052 TI - Surface recognition of a protein using designed transition metal complexes. AB - Each protein has a unique pattern of histidine residues on the surface. This paper describes the design, synthesis, and binding studies of transition metal complexes to target the surface histidine pattern of carbonic anhydrase (bovine erythrocyte). When the pattern of cupric ions on a complex matches the surface pattern of histidines of the protein, strong and selective binding can be achieved in aqueous buffer (pH = 7.0). The described method of protein recognition is applicable to proteins of known structures. With rapidly increasing number of solved protein structures, the method has wide applicability in purification, targeting, and sensing of proteins. PMID- 11427053 TI - Highly luminescent Cu(I)-phenanthroline complexes in rigid matrix and temperature dependence of the photophysical properties. AB - We synthesized new [Cu(NN)(2)](+)-type complexes where NN = 2-5 and denotes a 2,9 disubstituted-1,10-phenanthroline ligand (related complexes of 1 and 6 ligands are used for reference purposes). For 2, 3, and 4 the ligand substituents are long alkyl-type fragments, whereas in 5 a phenyl ring is directly attached to the chelating unit. At 298 K the four complexes display relatively intense metal-to ligand-charge-transfer (MLCT) emission bands with maxima around 720 nm, Phi(em) approximately 1 x 10(-)(3) and tau > 100 ns in deaerated CH(2)Cl(2). The emission behavior at 77 K in a CH(2)Cl(2)/MeOH matrix is quite different for complexes of alkyl- (2-4) versus phenyl-substituted (5) ligands. The former exhibit very intense emission bands centered around 642 nm and hypsochromically shifted with respect to 298 K, whereas the luminescence band of [Cu(5)(2)](+) is faint and shifted toward the infrared side. These results prompted us to study in detail the temperature dependence of luminescence properties of [Cu(2)(2)](+) and [Cu(5)(2)](+) in the 300-96 K range. For both complexes the excited state lifetimes increase monotonically by decreasing temperatures, and the trend is well described by an Arrhenius-type treatment involving two equilibrated MLCT excited levels. The emission bands show a similar behavior for the two compounds (intensity decrease and red-shift) only in the 300-120 K range, when the solvent is fluid. In the frozen regime (T CH(3)CN charge flow, but on the other hand the Natural population analysis indicates very little charge flow. For the Al-NH(3) complex, the polarization effect of NH(3) and the N-->Al sigma donation would enhance the dipole moment strength. However, there still remains a controversial disagreement between the theoretical predictions and the experimental results. Further experimental determination using the hexapole method for various metal-ligand complexes and clusters could reveal the basic nature of interaction in the complex systems in general, and this method would complement theoretical calculations. PMID- 11427063 TI - Ruthenium(II)-catalyzed isomer-selective cyclization of 1,6-dienes leading to exo methylenecyclopentanes: unprecedented cycloisomerization mechanism involving ruthenacyclopentane(hydrido) intermediate. AB - In the presence of a catalytic amount of ruthenium(II) complexes, [RuCl(2)(cod)](n)(), RuCl(2)(cod)(MeCN)(2), [RuCl(2)(nbd)](n)(), [RuCl(2)(CO)(3)](2), and Cp*Ru(cod)Cl, 1,6-dienes were effectively converted into the corresponding exo-methylenecyclopentanes in good to excellent yields with good isomer purity in i-PrOH at 90 degrees C. The alcoholic solvent was essential for the present catalytic cyclization, and the efficiency increased in the following order: t-BuOH << EtOH < or = i-PrOH. In contrast, a Ru(0) complex, (C(6)Me(6))Ru(cod), catalyzed the cycloisomerization only in 1,2-dichloroethane. The unusual isomer-selectivity occurred when a 1,7-octadiene was subjected to cyclization to give a similar exo-methylenecyclopentane isomer as the major product. The identical isomer selectivity was observed for the cyclization of unsymmetrical 1,6-dienes having one terminal- and one internal-alkene termini. On the basis of the results from the studies using the known ruthenium hydrides and deuterium-labeling substrates, the novel mechanism via the Ru(II) <--> Ru(IV) system involving a ruthenacyclopentane(hydrido) intermediate was proposed, which better explains the particular regiochemistry of the present cyclization than other previous mechanisms. PMID- 11427064 TI - Effects of molecular structure on the stability of a thermotropic liquid crystal. Gas electron diffraction study of the molecular structure of phenyl benzoate. AB - As a model of the core of molecules forming liquid crystals, the molecular structure of phenyl benzoate (Ph-C(=O)-O-Ph) at 409 K was determined by gas electron diffraction, and the relationship between the gas-phase structures of model compounds and the nematic-to-liquid transition temperatures was studied. Structural constraints were obtained from RHF/6-31G ab initio calculations. Vibrational mean amplitudes and shrinkage corrections were calculated from the harmonic force constants given by normal coordinate analysis. Thermal vibrations were treated as small-amplitude motions, except for the phenyl torsion, which was treated as a large-amplitude motion. The potential function for torsion was assumed to be V(phi(1),phi(2)) = V(12)(1 - cos 2phi(1))/2 + V(14)(1 - cos 4phi(1))/2 + V(22)(1 - cos 2phi(2))/2, where phi(1) and phi(2) denote the torsional angles around the C-Ph and O-Ph bonds, respectively. The potential constants (V(ij)()/kcal mol(-)(1)) and the principal structure parameters (r(g)/A, angle(alpha)/deg) with the estimated limits of error (3sigma) are as follows: V(12) = -1.3 (assumed); V(14) = -0.5(9); V(22) = 3.5(15); r(C=O) = 1.208(4); r(C(=O)-O) = 1.362(6); r(C(=O)-O) - r(O-C) = -0.044 (assumed); r(C(=O) C) = 1.478(10); = 1.396(1); angleOCO = 124.2(13); angleO=CC = 127.3(12); angleCOC = 121.4(22); ( angleOCC(cis) - angleOCC(trans))/2 = 3.0(15); ( angleC(=O)CC(cis) - angleC(=O)CC(trans))/2 = 4.8(17), where < > means an average value and C-C(cis) and C-C(trans) bonds are cis and trans to the C(=O)-O bond, respectively. The torsional angle around the O-Ph bond was determined to be 64(+26,-12) degrees. An apparent correlation was found between the contributions of the cores to the clearing point of liquid crystals and the gas-phase structures of model compounds of the cores of mesogens, i.e., phenyl benzoate, trans-azobenzene (t-AB), N-benzylideneaniline, N-benzylideneaniline N-oxide (NBANO), trans-azoxybenzene (t-AXB), and trans-stilbene. The structures of t-AB, NBANO, and t-AXB have been obtained by our research group. PMID- 11427065 TI - Chemistry on surface-confined molecules: an approach to anchor isolated functional units to surfaces. AB - The synthesis of surface-confined, nanometer-sized dendrimers and Au nanoparticles was performed starting from single Pd(II) pincer adsorbate molecules (10) embedded as isolated species into 11-mercapto-1-undecanol and decanethiol self-assembled monolayers (SAMs) on gold. The coordination of monolayer-protected Au nanoclusters (MPCs) bearing phosphine moieties at the periphery (13), or dendritic wedges (8) having a phosphine group at the focal point, to SAMs containing individual Pd(II) pincer molecules was monitored by tapping mode atomic force microscopy (TM AFM). The individual Pd(II) pincer molecules embedded in the decanethiol SAM were visualized by their coordination to phosphine MPCs 13; isolated objects with a height of 3.5 +/- 0.7 nm were observed by TM AFM. Reaction of these embedded Pd(II) pincer molecules with the dendritic wedge 8 yielded individual molecules with a height of 4.3 +/- 0.2 nm. PMID- 11427066 TI - Origin of diastereofacial selectivity in tertiary 2-adamantyl cations. AB - The intrinsic factors governing the diastereofacial selectivity of 2-methyl-5-X-2 adamantyl cations (X = F (I(F)), Si(CH(3))(3) (I(Si))) toward a representative nucleophile, i.e., methanol, have been investigated in the gas phase at 750 Torr and in the 20-80 degrees C temperature range. The kinetic results indicate that CH(3)OH addition to I(F) proceeds through tight transition structures (TS(F)(syn) and TS(F)(anti)) characterized by advanced C-O bonding. The same interactions are much less pronounced in the comparatively loose transition structures involved in the CH(3)OH addition to I(Si) (TS(Si)(syn) and TS(Si)(anti)). The experimental evidence indicates that the activation barriers for the anti addition to I(F) and I(Si) are invariably lower than those for the syn attack. Large adverse entropic factors account for the preferred syn diastereoselectivity observed in the reaction with I(F). Entropy plays a minor role in the much looser transition structures involved in the reaction with I(Si), which instead exhibits a preferred anti diastereoselectivity. Comparison of the above gas-phase results with related theoretical and solution data suggests that the diastereofacial selectivity of I(F) and I(Si) measured in solution arises in part from the differential solvation of the two faces of the pyramidalized ions. PMID- 11427067 TI - Mechanism of activation of an immunosuppressive drug: azathioprine. Quantum chemical study on the reaction of azathioprine with cysteine. AB - Azathioprine is an important drug used in the therapy of autoimmune disorders and in preventing graft rejection. Its molecule is composed of two main moieties: mercaptopurine and imidazole derivative. It is an immunosuppressive agent whose biological activity results from its in vivo mercaptolysis mediated by a nucleophilic attack on the C(5i) atom of imidazole ring of the azathioprine molecule. Solvation model SM5.4 with the PM3 Hamiltonian have been applied to model the reaction of azathioprine with cysteine. The employed quantum mechanical method shed new light on the mechanism of the reaction of azathioprine with cysteine in aqueous solution. The obtained results indicated that the first step in the reaction most likely involves the nucleophilic attack of the COO(-) of cysteine on the C(5i) atom of the imidazole ring of azathioprine, followed by a subsequent intramolecular attack of the SH group of the cysteine residue. It was shown that biogenic thiols such as glutathione or cysteine facilitate the first and crucial step of azathioprine metabolism, due to the presence of COO(-), SH, and NH(3)(+) groups in their molecules. PMID- 11427068 TI - Monooxygenation mechanism by cytochrome p-450. AB - The substrate oxygenation mechanism by an ultimate species in monooxygenation by cytochrome P-450 (compound I) was investigated by the density functional theory method. An initial model compound was constructed from a structure obtained by 300-ps molecular dynamics simulation of compound I-formed P-450cam under physiologic conditions, and it consisted of porphine for protoporphyrin IX, S(-) CH(3) for the side chain of Cys357 of the fifth ligand of heme, a methane molecule for the substrate, a heme iron, and an oxygen atom of the sixth ligand of heme. The results of the calculation revealed that the substrate oxygenation mechanism had four elementary processes, i.e., (1) formation of [FeOH](3+) and a substrate radical by hydrogen atom abstraction from the substrate caused by [FeO](3+), (2) rotation of the OH group of the sixth ligand of [FeOH](3+) produced by process 1, (3) substrate radical binding with the [FeOH](3+), and (4) elimination of the oxygenated substrate formed at the sixth ligand binding site. The rate-determining step is process 1, hydrogen atom abstraction from the substrate, and the activation energy was determined to be about 15 kcal/mol. For this reason, it is thought that this reaction occurs in vivo. PMID- 11427069 TI - Water-soluble conjugated oligomers: effect of chain length and aggregation on photoluminescence-quenching efficiencies. PMID- 11427070 TI - Deoxygenations of (silox)(3)WNO and R(3)PO by (silox)(3)M (M = V, Ta) and (silox)(3)NbL (silox = (t)Bu(3)SiO): consequences of electronic effects. PMID- 11427072 TI - A stable five-coordinate platinum(IV) alkyl complex. PMID- 11427071 TI - 1,3,5-Tricyano-2,4,6-tris(vinyl)benzene derivatives with large second-order nonlinear optical properties. PMID- 11427073 TI - Five-coordinate trispyrazolylborate dihydridosilyl platinum(IV) complexes. PMID- 11427074 TI - Structural studies of Sc metallofullerenes by high-resolution ion mobility measurements. PMID- 11427075 TI - Selective C-C bond activation of methyl pyruvate on Ni(111) to yield surface methoxycarbonyl. PMID- 11427076 TI - One-electron reduction of an antiferromagnetically coupled triradical yields a mixed-valent biradical with enhanced ferromagnetic coupling. PMID- 11427077 TI - A general strategy for absolute stereochemical control in enone-olefin [2+2] photocycloaddition reactions. PMID- 11427078 TI - Oxyindolophyrin: a novel fluoride receptor derived from N-confused corrole isomer. PMID- 11427079 TI - Redirection of oxidation reactions by a polyoxomolybdate: oxydehydrogenation instead of oxygenation of alkanes with tert-butylhydroperoxide in acetic acid. PMID- 11427081 TI - Stereoselective synthesis of isomeric functionalized 1,3-dienes from cyclobutenones. PMID- 11427080 TI - Fluoride-free cross-coupling of organosilanols. PMID- 11427082 TI - Dimeric triarylbismuthane oxide: a novel efficient oxidant for the conversion of alcohols to carbonyl compounds. PMID- 11427083 TI - Direct measurement of enol ether radical cation reaction kinetics. PMID- 11427084 TI - Excited-state dynamics of oligo(p-phenyleneethynylene): quadratic coupling and torsional motions. PMID- 11427085 TI - Mechanistic studies on phosphopantothenoylcysteine decarboxylase. PMID- 11427086 TI - Polar interactions with aromatic side chains in alpha-helical peptides: Ch...O H bonding and cation-pi interactions. PMID- 11427088 TI - Steric stabilization of homoleptic bis(pi-allyl) complexes of chromium(II) and iron(II). PMID- 11427087 TI - Mellitic triimides: C(3)-symmetric, three-electron acceptors for supramolecular chemistry. PMID- 11427089 TI - Gamma-butyrolactone synthesis via catalytic asymmetric cyclocarbonylation. PMID- 11427090 TI - Molecular recognition of DNA intercalators at nanomolar concentration in water. PMID- 11427091 TI - High-throughput synthesis of nanoscale materials: structural optimization of functionalized one-step star polymers. PMID- 11427094 TI - What is goal blood pressure for the treatment of hypertension? PMID- 11427095 TI - Low-molecular-weight heparins in the treatment of acute coronary syndromes. AB - Platelet aggregation and activation of coagulation are key events in the development of acute coronary syndromes. Patients with an acute coronary syndrome are at high risk of death or myocardial infarction, and hence there is a strong rationale for the use of antithrombotic agents. Heparin has been shown to reduce the risk of death or myocardial infarction in aspirin-treated patients with acute coronary syndromes, but it has a number of limitations, including the need for regular monitoring and the risk of hemorrhage and thrombocytopenia. Low-molecular weight heparins offer a number of practical and clinical advantages over unfractionated heparin, such as higher bioavailability and administration by subcutaneous injection. Several low-molecular-weight heparins are available that differ in their biochemical and pharmacologic properties, and it is not possible to predict their clinical efficacy from their pharmacologic profile. The decision regarding the use of a specific low-molecular-weight heparin should be based on the efficacy and safety data available for each product. In clinical trials comparing low-molecular-weight heparin with heparin, only enoxaparin sodium has been shown to reduce the risk of coronary events in patients with non-ST segment elevation acute coronary ischemia. PMID- 11427096 TI - Angiotensin II subtype 1 receptor blockers and renal function. AB - Blood pressure reduction is the most significant factor in delaying onset and progression of renal disease. Blockade of the renin-angiotensin system (RAS) using angiotensin-converting enzyme inhibitors (ACEIs) delays renal disease progression. More recently, agents that block the RAS by preventing angiotensin II from binding to its subtype 1 receptor (ARBs) have been developed in an effort to prevent deleterious consequences of pathologic levels of angiotensin II and to reduce the adverse effects of RAS blockade associated with ACEIs. Human studies with a variety of ARBs have clearly demonstrated the antihypertensive and antiproteinuric efficacy of these agents in patients with progressive renal diseases. Moreover, the effects of ARBs are similar or identical to those of ACEIs. Ongoing long-term clinical trials are designed to determine whether ARBs also preserve renal function similar to ACEIs. Specifically, the role of ARBs in patients with hypertension and type 2 diabetes is being evaluated in 3 large trials, including Appropriate Blood Pressure Control in Diabetes-Part 2 With Valsartan, the Losartan Renal Protection Study, and the Irbesartan Diabetic Nephropathy Trial. Definitive evidence of the long-term protective effects of ARBs in chronic progressive renal disease is expected from these important studies. PMID- 11427097 TI - Relationship of blood pressure to 25-year mortality due to coronary heart disease, cardiovascular diseases, and all causes in young adult men: the Chicago Heart Association Detection Project in Industry. AB - BACKGROUND: Data are limited on blood pressure (BP) in young adults and long-term mortality. Moreover, screening and hypertension treatment guidelines have been based mainly on findings for middle-aged and older populations. This study assesses relationships of BP measured in young adult men to long-term mortality due to coronary heart disease (CHD), cardiovascular diseases (CVD), and all causes. METHODS: This cohort from the Chicago Heart Association Detection Project in Industry included 10 874 men aged 18 to 39 years at baseline (1967-1973), not receiving antihypertensive drugs, and without CHD or diabetes. Relationship of baseline BP to 25-year CHD, CVD, and all-cause mortality was assessed. RESULTS: Age-adjusted association of systolic BP to CHD mortality was continuous and graded. Multivariate-adjusted CHD hazard ratios (HRs) for 1 SD higher systolic BP (15 mm Hg) and diastolic BP (10 mm Hg) were 1.26 (95% confidence interval [CI], 1.11-1.44) and 1.17 (95% CI, 1.01-1.35), respectively. Compared with the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure stratum with normal BP (and lowest mortality rates), the large strata with high-normal BP and stage 1 hypertension had 25-year absolute risks for death of 63 and 72 per 1000, respectively, and absolute excess risks of 10 and 20 per 1000, respectively; accounted for 59.8% of all excess CHD, CVD, and all-cause mortality; and were estimated to have life expectancy shortened by 2.2 and 4.1 years, respectively. CONCLUSIONS: In young adult men, BP above normal was significantly related to increased long-term mortality due to CHD, CVD, and all causes. Population-wide primary prevention, early detection, and control of higher BP are indicated from young adulthood on. PMID- 11427098 TI - Cardiac arrest in medical and dental practices: implications for automated external defibrillators. AB - BACKGROUND: To determine the need for placing automated external defibrillators (AEDs) in medical and dental practices, we identified cardiac arrests at these locations. METHODS: Locations of cardiac arrest were abstracted from Emergency Medical Services data from January 1, 1990, through December 31, 1996. We calculated the annual incidence of cardiac arrest per type of practice. RESULTS: There were 142 cardiac arrests in medical or dental practices. Dialysis centers had a relatively high incidence of cardiac arrest (>/=0.746 per practice annually). Cardiology, internal and family medicine, and urgent care centers had a medium incidence (>/=0.01 per practice annually). All other medical and dental practices had a low incidence (/=5 apnea and hypopnea episodes per hour of sleep). Prospective analyses showed that persons with chronic severe nasal congestion had a high risk of habitual snoring according to the data from the 5-year follow-up survey: the odds ratio for habitual snoring and reporting congestion always or almost always at both baseline and follow-up was 4.9 (95% confidence interval, 2.8-8.8). CONCLUSIONS: Nocturnal nasal congestion is a strong independent risk factor for habitual snoring, including snoring without frank sleep apnea. Intervention studies are needed to determine if snoring can be reduced with treatment of nasal congestion. PMID- 11427100 TI - Temporal trends (1986-1997) in cholesterol level assessment and management practices in patients with acute myocardial infarction: a population-based perspective. AB - BACKGROUND: Elevated serum cholesterol levels are associated with increased risk for acute myocardial infarction (AMI) and adverse patient outcomes. It is unclear what proportion of patients have their serum cholesterol levels measured during hospitalization for AMI and are given hypolipidemic therapy. OBJECTIVE: To examine decade-long trends in measurement of serum cholesterol levels during hospitalization for AMI and use of hypolipidemic therapy. METHODS: Observational study of 5204 residents of the Worcester, Mass, metropolitan area hospitalized with validated AMI in all greater Worcester hospitals in seven 1-year periods from 1986 through 1997. RESULTS: Increases in the measurement of serum cholesterol levels during hospitalization for AMI were observed between 1986 and 1991, followed by a progressive decrease; only 24% of patients with AMI in 1997 underwent cholesterol level testing. Younger age, male sex, and absence of a history of cardiovascular disease were associated with an increased likelihood measurement of serum cholesterol levels. Although the relative use of hypolipidemic therapy increased significantly over time (0.4% in 1986 vs 10.7% in 1997), the absolute rate of use remained low. In patients with elevated serum cholesterol levels (>/=6.2 mmol/L [>/=240 mg/dL]), 1.9% received hypolipidemic therapy in 1986 and 36.6% in 1997. CONCLUSIONS: These findings suggest recent declines in the assessment of total cholesterol levels in patients hospitalized with AMI. Although the use of hypolipidemic therapy during hospitalization for AMI has increased over time, considerable room for improvement remains. PMID- 11427101 TI - A meta-analysis of salvage therapy for Pneumocystis carinii pneumonia. AB - OBJECTIVE: To determine the relative efficacies of alternative antipneumocystis agents in human immunodeficiency virus (HIV)-infected patients with Pneumocystis carinii pneumonia unresponsive to primary drug treatment with a combination product of trimethoprim and sulfamethoxazole or parenteral pentamidine. METHODS: Meta-analysis of 27 published clinical drug trials, case series, and case reports involving P carinii pneumonia. Data extracted included underlying disease, primary antipneumocystis treatment, days of failed primary treatment, salvage regimen, use of systemic corticosteroids and antiretroviral drugs, and clinical outcome. RESULTS: In 497 patients with microbiologically confirmed P carinii pneumonia (456 with HIV or acquired immunodeficiency syndrome), initial antipneumocystis treatment failed and they therefore required alternative drug therapy. Failed regimens included trimethoprim-sulfamethoxazole (160 patients), intravenous pentamidine (63 patients), trimethoprim-sulfamethoxazole and/or pentamidine (258 patients), aerosolized pentamidine (6 patients), atovaquone (3 patients), dapsone (3 patients), a combination product of trimethoprim and dapsone (2 patients), and trimethoprim-sulfamethoxazole followed by a combination of clindamycin and primaquine phosphate (2 patients). Efficacies of salvage regimens were as follows: clindamycin-primaquine (42 to 44 [88%-92%] of 48 patients; P<10(-8)), atovaquone (4 [80%] of 5), eflornithine hydrochloride (40 [57%] of 70; P<.01), trimethoprim-sulfamethoxazole (27 [53%] of 51; P<.08), pentamidine (64 [39%] of 164), and trimetrexate (47 [30%] of 159). CONCLUSION: The combination of clindamycin plus primaquine appears to be the most effective alternative treatment for patients with P carinii pneumonia who are unresponsive to conventional antipneumocystis agents. PMID- 11427102 TI - Increased plasma methylmalonic acid level does not predict clinical manifestations of vitamin B12 deficiency. AB - BACKGROUND: The prevalence of vitamin B(12) deficiency, defined as an elevated concentration of plasma methylmalonic acid (P-MMA), has been estimated to be 15% to 44% in the elderly. However, we do not know whether an increased P-MMA level actually indicates or predicts a clinical condition in need of treatment. PARTICIPANTS AND METHODS: In a follow-up study, 432 individuals not treated with vitamin B(12) were examined 1.0 to 3.9 years after initial observation of an increased P-MMA concentration (>0.28 micromol/L). The examination included laboratory tests, a structured interview to disclose symptoms, a food frequency questionnaire, and a clinical examination including a Neurological Disability Score. RESULTS: Variation in P-MMA levels over time was high (coefficient of variation, 34%). In only 16% of participants, P-MMA levels increased substantially, whereas 44% showed a decrease. Level of P-MMA was significantly but not strongly associated with levels of plasma cobalamins (r = -0.22, P<.001) and plasma total homocysteine (r = 0.37, P<.001). After adjustment for age and sex, we found no associations between P-MMA concentration and the total symptom score (P =.61), the total Neurological Disability Score (P =.64), or other clinical manifestations related to vitamin B(12) deficiency. CONCLUSIONS: An increased level of P-MMA did not predict a further increase with time and clinical manifestations related to vitamin B(12) deficiency. We therefore challenge the use of an increased P-MMA concentration as the only marker for diagnosis of vitamin B(12) deficiency. PMID- 11427103 TI - Physical activity and television watching in relation to risk for type 2 diabetes mellitus in men. AB - BACKGROUND: Television (TV) watching, a major sedentary behavior in the United States, has been associated with obesity. We hypothesized that prolonged TV watching may increase risk for type 2 diabetes. METHODS: In 1986, 37 918 men aged 40 to 75 years and free of diabetes, cardiovascular disease, and cancer completed a detailed physical activity questionnaire. Starting from 1988, participants reported their average weekly time spent watching TV on biennial questionnaires. RESULTS: A total of 1058 cases of type 2 diabetes were diagnosed during 10 years (347 040 person-years) of follow-up. After adjustment for age, smoking, alcohol use, and other covariates, the relative risks (RRs) for type 2 diabetes across increasing quintiles of metabolic equivalent hours (MET-hours) per week were 1.00, 0.78, 0.65, 0.58, and 0.51 (P for trend, <.001). Time spent watching TV was significantly associated with higher risk for diabetes. After adjustment for age, smoking, physical activity levels, and other covariates, the RRs of diabetes across categories of average hours spent watching TV per week (0-1, 2-10, 11-20, 21-40, and >40) were 1.00, 1.66, 1.64, 2.16, and 2.87, respectively (P for trend, <.001). This association was somewhat attenuated after adjustment for body mass index, but a significant positive gradient persisted (RR comparing extreme categories, 2.31; P for trend,.01). CONCLUSIONS: Increasing physical activity is associated with a significant reduction in risk for diabetes, whereas a sedentary lifestyle indicated by prolonged TV watching is directly related to risk. Our findings suggest the importance of reducing sedentary behavior in the prevention of type 2 diabetes. PMID- 11427104 TI - Quality of care for hospitalized medicare patients at risk for pressure ulcers. AB - BACKGROUND: No state peer review organization has attempted to identify processes of care related to pressure ulcer prediction and prevention in US hospitals. OBJECTIVE: To profile and evaluate the processes of care for Medicare patients hospitalized at risk for pressure ulcer development by means of the Medicare Quality Indicator System pressure ulcer prediction and prevention module. METHODS: A multicenter retrospective cohort study with medical record abstraction was used to obtain a total of 2425 patients aged 65 years and older discharged from acute care hospitals after treatment for pneumonia, cerebrovascular disease, or congestive heart failure. Six processes of care for prevention of pressure ulcers were evaluated: use of daily skin assessment; use of a pressure-reducing device; documentation of being at risk; repositioning for a minimum of 2 hours; nutritional consultation initiated for patients with nutritional risk factors; and staging of pressure ulcer. The associations between processes of care and incidence of pressure ulcer were determined with Kaplan-Meier survival analyses. RESULTS: National estimates of compliance with process of care were as follows: use of daily skin assessment, 94%; use of pressure-reducing device, 7.5%; documentation of being at risk, 22.6%; repositioning for a minimum of 2 hours, 66.2%; nutritional consultation, 34.3%; stage 1 pressure ulcer staged, 20.2%; and stage 2 or greater ulcer staged, 30.9%. CONCLUSION: These results suggest that US hospitals and physicians have numerous opportunities to improve care related to pressure ulcer prediction and prevention. PMID- 11427105 TI - Primary cerebellar degeneration and HIV. PMID- 11427106 TI - Erythema multiforme associated with bupropion use. PMID- 11427107 TI - Causes of death in patients with systemic lupus erythematosus in Sao Paulo, Brazil: a study of 113 autopsies. PMID- 11427108 TI - Consensus needed for evaluating safe and adequate anticoagulant control. PMID- 11427109 TI - Chronic fatigue syndrome, decreased exercise capacity, and adrenal insufficiency. PMID- 11427111 TI - JAMA 100 Years Ago: HYSTERICAL SENSORY APHASIA-WORD-DEAFNESS-IN A CHILD. PMID- 11427112 TI - Quick Uptakes: Expanding Care Information. PMID- 11427113 TI - Quick Uptakes: Emergency Care of Children Improved. PMID- 11427115 TI - Quick Uptakes: beta-Blocker Benefit. PMID- 11427114 TI - Quick Uptakes: Predicting Brain Cell Death. PMID- 11427117 TI - Physiological neuroimaging: emerging clinical applications. PMID- 11427118 TI - Confidentiality and privacy of electronic medical records: psychiatrists explore risks of the "information age". PMID- 11427119 TI - Study indicates utility for new breast cancer prognostic marker. PMID- 11427120 TI - What lures women physicians to practice medicine in rural areas? PMID- 11427126 TI - Relapse of depression after electroconvulsive therapy. PMID- 11427127 TI - Relapse of depression after electroconvulsive therapy. PMID- 11427129 TI - Relationship between postmenopausal hormone replacement therapy and ovarian cancer. PMID- 11427130 TI - Relationship between postmenopausal hormone replacement therapy and ovarian cancer. PMID- 11427132 TI - Breast cancer in women with HIV/AIDS. PMID- 11427134 TI - A novel about bioterrorism. PMID- 11427137 TI - Pervasive developmental disorders in preschool children. AB - CONTEXT: Prevalence rates of autism-spectrum disorders are uncertain, and speculation that their incidence is increasing continues to cause concern. OBJECTIVE: To estimate the prevalence of pervasive developmental disorders (PDDs) in a geographically defined population of preschool children. DESIGN, SETTING, AND PARTICIPANTS: Survey conducted July 1998 to June 1999 in Staffordshire, England. The area's 15 500 children aged 2.5 to 6.5 years were screened for developmental problems. Children with symptoms suggestive of a PDD were intensively assessed by a multidisciplinary team, which conducted standardized diagnostic interviews and administered psychometric tests. MAIN OUTCOME MEASURE: Prevalence estimates for subtypes of PDDs. RESULTS: A total of 97 children (79.4% male) were confirmed to have a PDD. The prevalence of PDDs was estimated to be 62.6 (95% confidence interval, 50.8-76.3) per 10 000 children. Prevalences were 16.8 per 10 000 for autistic disorder and 45.8 per 10 000 for other PDDs. The mean age at diagnosis was 41 months, and 81% were originally referred by health visitors (nurse specialists). Of the 97 children with a PDD, 25.8% had some degree of mental retardation and 9.3% had an associated medical condition. CONCLUSIONS: Our results suggest that rates of PDD are higher than previously reported. Methodological limitations in existing epidemiological investigations preclude interpretation of recent high rates as indicative of increased incidence of these disorders although this hypothesis requires further rigorous testing. Attention is nevertheless drawn to the important needs of a substantial minority of preschool children. PMID- 11427138 TI - Effect of condoms on reducing the transmission of herpes simplex virus type 2 from men to women. AB - CONTEXT: Herpes simplex virus type 2 (HSV-2) is one of the most common sexually transmitted infections in the United States. No prospective study has shown the ability of condoms to reduce transmission of HSV-2. OBJECTIVE: To evaluate risk factors for HSV-2 acquisition and efficacy of condoms in prevention of HSV-2 transmission. DESIGN: Analysis of data from a randomized, double-blind, placebo controlled trial conducted December 13, 1993, to June 28, 1996, of an ineffective candidate HSV-2 vaccine with 18 months of follow-up. SETTING: Eighteen clinical trial centers in the United States. PARTICIPANTS: A total of 528 monogamous couples discordant for HSV-2 infection, including an HSV-2-susceptible population of 261 men and 267 women. MAIN OUTCOME MEASURE: Acquisition of HSV-2 infection by susceptible partners, compared with those remaining free of HSV-2 with regard to demographic characteristics, sexual activity, and condom use. RESULTS: Twenty-six women (9.7%) vs 5 men (1.9%) acquired HSV-2, for a rate per 10 000 sex acts (episodes of sexual intercourse) of 8.9 vs 1.5, respectively (P<.001). In multivariable analysis, younger age (adjusted hazard ratio [HR] per 5 years, 1.57; 95% confidence interval [CI], 1.22-2.04), seropositivity for HSV-1 and HSV 2 vs HSV-2 alone in the source partner (adjusted HR, 2.34; 95% CI, 1.14-4.82), and more frequent sexual activity (adjusted HR per additional sex act per week, 1.10; 95% CI, 1.01-1.19) were associated with higher risk of HSV-2 acquisition. Condom use during more than 25% of sex acts was associated with protection against HSV-2 acquisition for women (adjusted HR, 0.085; 95% CI, 0.01-0.67) but not for men (adjusted HR, 2.02; 95% CI, 0.32-12.50). Risk of HSV-2 transmission declined from 8.5 per 100 person-years in the initial 150-day interval to 0.9 per 100 person-years in the final 150-day interval (P =.002 for trend), concurrent with a decrease in sexual activity and proportion of sex acts occurring when the source partner had genital lesions. CONCLUSIONS: Condom use offers significant protection against HSV-2 infection in susceptible women. Changes in sexual behavior, correlated with counseling about avoiding sex when a partner has lesions, were associated with reduction in HSV-2 acquisition over time. These data suggest that identification of discordant couples can reduce transmission of HSV-2, especially for heterosexual couples in which the male partner has HSV-2 infection. PMID- 11427139 TI - Policy analysis of cervical cancer screening strategies in low-resource settings: clinical benefits and cost-effectiveness. AB - CONTEXT: Cervical cancer is a leading cause of cancer-related death among women in developing countries. In such low-resource settings, cytology-based screening is difficult to implement, and less complex strategies may offer additional options. OBJECTIVE: To assess the cost-effectiveness of several cervical cancer screening strategies using population-specific data. DESIGN AND SETTING: Cost effectiveness analysis using a mathematical model and a hypothetical cohort of previously unscreened 30-year-old black South African women. Screening tests included direct visual inspection (DVI) of the cervix, cytologic methods, and testing for high-risk types of human papillomavirus (HPV) DNA. Strategies differed by number of clinical visits, screening frequency, and response to a positive test result. Data sources included a South African screening study, national surveys and fee schedules, and published literature. MAIN OUTCOME MEASURES: Years of life saved (YLS), lifetime costs in US dollars, and incremental cost-effectiveness ratios (cost per YLS). RESULTS: When analyzing all strategies performed as a single lifetime screen at age 35 years compared with no screening, HPV testing followed by treatment of screen-positive women at a second visit, cost $39/YLS (27% cancer incidence reduction); DVI, coupled with immediate treatment of screen-positive women at the first visit was next most effective (26% cancer incidence reduction) and was cost saving; cytology, followed by treatment of screen-positive women at a second visit was least effective (19% cancer incidence reduction) at a cost of $81/YLS. For any given screening frequency, when strategies were compared incrementally, HPV DNA testing generally was more effective but also more costly than DVI, and always was more effective and less costly than cytology. When comparing all strategies simultaneously across screening frequencies, DVI was the nondominated strategy up to a frequency of every 3 years (incremental cost-effectiveness ratio, $460/YLS), and HPV testing every 3 years (incremental cost-effectiveness ratio, $11 500/YLS) was the most effective strategy. CONCLUSION: Cervical cancer screening strategies that incorporate DVI or HPV DNA testing and eliminate colposcopy may offer attractive alternatives to cytology-based screening programs in low-resource settings. PMID- 11427140 TI - Relationship between annual volume of patients treated by admitting physician and mortality after acute myocardial infarction. AB - CONTEXT: Acute myocardial infarction (AMI) is a common condition that is treated by physicians with varying levels of clinical experience, but whether the level of experience affects outcome remains uncertain. OBJECTIVE: To evaluate the relationship between the average annual volume of cases treated by admitting physicians and mortality after AMI. DESIGN, SETTING, AND PATIENTS: Retrospective cohort study using linked administrative databases containing patient admission information for 98 194 patients treated by 5374 physicians between April 1, 1992, and March 31, 1998, in Ontario, Canada. MAIN OUTCOME MEASURES: Mortality risk rates for 30 days and 1 year post-AMI, adjusted by physician volume and patient, physician, and hospital characteristics. RESULTS: The 30-day mortality rate was 13.5% and the 1-year mortality rate was 21.8%. A strong inverse relationship between the average annual volume of AMI cases treated by the admitting physician and mortality after an AMI was observed. The 30-day risk-adjusted mortality rate was 15.3% for physicians who treated 5 or fewer AMI cases per year (lowest quartile) compared with 11.8% for physicians who treated more than 24 AMI cases annually (highest quartile; P<.001). The 1-year risk-adjusted mortality rate was 24.2% for physicians who treated 5 or fewer AMI cases per year (lowest quartile) compared with 19.6% for physicians who treated more than 24 AMI cases annually (highest quartile; P<.001). CONCLUSION: Patients with AMI who are treated by high volume admitting physicians are more likely to survive at 30 days and 1 year. PMID- 11427141 TI - Involvement in caregiving and adjustment to death of a spouse: findings from the caregiver health effects study. AB - CONTEXT: Most deaths in the United States occur among older persons who have 1 or more disabling conditions. As a result, many deaths are preceded by an extended period during which family members provide care to their disabled relative. OBJECTIVE: To better understand the effect of bereavement on family caregivers by examining predeath vs postdeath changes in self-reported and objective health outcomes among elderly persons providing varying levels of care prior to their spouse's death. DESIGN AND SETTING: Prospective, population-based cohort study conducted in 4 US communities between 1993 and 1998. PARTICIPANTS: One hundred twenty-nine individuals aged 66 to 96 years whose spouse died during an average 4 year follow-up. Individuals were classified as noncaregivers (n = 40), caregivers who reported no strain (n = 37), or strained caregivers (n = 52). MAIN OUTCOME MEASURES: Changes in depression symptoms (assessed by the 10-item Center for Epidemiological Studies-Depression [CES-D] scale), antidepressant medication use, 6 health risk behaviors, and weight among the 3 groups of participants. RESULTS: Controlling for age, sex, race, education, prevalent cardiovascular disease at baseline, and interval between predeath and postdeath assessments, CES-D scores remained high but did not change among strained caregivers (9.44 vs 9.19; P =.76), while these scores increased for both noncaregivers (4.74 vs 8.25; F(1,116) = 14.33; P<.001) and nonstrained caregivers (4.94 vs 7.13; F(1,116) = 4.35; P =.04). Noncaregivers were significantly more likely to be using nontricyclic antidepressant medications following the death than the nonstrained caregiver group (odds ratio [OR], 12.85; 95% confidence interval [CI], 1.02 162.13; P =.05). The strained caregiver group experienced significant improvement in health risk behaviors following the death of their spouse (1.47 vs 0.66 behaviors; F(1,118) = 20.23; P<.001), while the noncaregiver and nonstrained caregiver groups showed little change (0.27 vs 0.27 [P =.99] and 0.46 vs 0.27 [P =.39] behaviors, respectively). Noncaregivers experienced significant weight loss following the death (149.1 vs 145.3 lb [67.1 vs 65.4 kg]; F(1,101) = 8.12; P =.005), while the strained and nonstrained caregiving groups did not show significant weight change (156.2 vs 155.2 lb [70.3 vs 69.8 kg] [P =.41] and 156.2 vs 154.0 lb [70.3 vs 69.3 kg] [P =.12], respectively). CONCLUSIONS: These data indicate that the impact of losing one's spouse among older persons varies as a function of the caregiving experiences that precede the death. Among individuals who are already strained prior to the death of their spouse, the death itself does not increase their level of distress. Instead, they show reductions in health risk behaviors. Among noncaregivers, losing one's spouse results in increased depression and weight loss. PMID- 11427142 TI - Preimplantation diagnosis for Fanconi anemia combined with HLA matching. AB - CONTEXT: The advent of single-cell polymerase chain reaction (PCR) has presented the opportunity for combined preimplantation genetic diagnosis (PGD) and HLA antigen testing. This is a novel and useful way to preselect a potential donor for an affected sibling requiring stem cell transplantation. OBJECTIVE: To perform in vitro fertilization (IVF) and preimplantation HLA matching combined with PGD for Fanconi anemia (FA). DESIGN: DNA analysis for the IVS 4 + 4 A-->T (adenine to thymine) mutation in the FA complement C (FANCC) gene in single blastomeres, obtained by biopsy of embryos, to identify genetic status and HLA markers of each embryo before intrauterine transfer. SETTING: In vitro fertilization programs at large medical centers in Chicago, Ill, and Denver, Colo. PARTICIPANTS: A couple, both carriers of the IVS 4 + 4 A-->T mutation in the FANCC gene with an affected child requiring an HLA-compatible donor for cord blood transplantation. MAIN OUTCOME MEASURES: DNA analysis of single blastomeres to preselect unaffected embryos representing an HLA match for the affected sibling. RESULTS: Of 30 embryos tested in 4 IVF attempts, 6 were homozygous affected and 24 were unaffected. Five of these embryos were also found to be HLA compatible, of which 2 were transferred in the first and 1 in each of the other 3 cycles, resulting in a pregnancy and birth of an unaffected child in the last cycle. CONCLUSION: To our knowledge, this is the first PGD with HLA matching, demonstrating feasibility of preselecting unaffected embryos that can also be an HLA-compatible source for stem cell transplantation for a sibling. PMID- 11427143 TI - A 40-year-old woman who noticed a medication error. PMID- 11427144 TI - A 26-year-old woman with shoulder pain, 1 year later. PMID- 11427145 TI - Pervasive developmental disorders in young children. PMID- 11427146 TI - Ethical implications of a new application of preimplantation diagnosis. PMID- 11427152 TI - Cellular basis for paracrine regulation of ovarian follicle development. AB - Paracrine factors secreted by oocytes and somatic cells regulate many important aspects of early ovarian follicle development in mammals. From activation of dormant primordial follicles to selection of secondary follicles, locally acting factors have been identified that appear to exert important effects on the growth and differentiation of oocytes and granulosa cells. This article summarizes evidence to support a model for bi-directional paracrine communication that is based on developmental regulation of the delivery and reception of paracrine factors at the oocyte-granulosa cell interface. Transzonal projections that originate from granulosa cells and terminate at the oocyte plasma membrane provide a polarized means to orient the secretory organelles of somatic cells. Characterization of transzonal projections in follicles from normal and genetically modified mice reveals dynamic changes in the density and stability of transzonal projections. On the basis of new data analysing the orientation and cytoskeletal content of transzonal projections in mammalian oocytes, a model is proposed for regulation of paracrine growth factor secretion by follicle stimulating hormone. These findings have immediate implications for ovarian hyperstimulation protocols and follicle culture models as related to the production of mammalian embryos by assisted reproductive technologies. PMID- 11427153 TI - Aneuploidy in human spermatozoa: FISH analysis in men with constitutional chromosomal abnormalities, and in infertile men. AB - Reproductive difficulties are associated intimately with cytogenetic abnormalities. This article reviews multicolour fluorescence in situ hybridization studies on spermatozoa from men with constitutional chromosomal abnormalities and the consequences for spermatozoa, and on chromosomal abnormalities in the spermatozoa of infertile men who have normal somatic karyotypes. In 47,XYY men, the frequencies of 24,XY and 24,YY spermatozoa appear to be < or = 1%. Klinefelter (47,XXY) and mosaic Klinefelter patients had sperm aneuploidy frequencies of 2-25% and 1.5-7.0%, respectively. Robertsonian translocation carriers had 3-27% spermatozoa unbalanced for the chromosomes involved in the translocation, with a possible modest interchromosomal effect, but none of the increased frequencies of chromosomal disomy approached 1%. The frequency of chromosomally unbalanced spermatozoa in reciprocal translocations averages 50%, is strongly dependent on the chromosomes involved in the individual translocation, and may be slightly increased as a result of a small interchromosomal effect. Infertile men with a normal karyotype and low sperm concentration or certain types of morphologically abnormal spermatozoa have a significantly increased risk of producing aneuploid spermatozoa, particularly for the sex chromosomes. An increased risk of sperm aneuploidy was not observed in infertile men with poor sperm motility or in those with a normal karyotype and normal semen parameters. PMID- 11427154 TI - Roles of activin and its signal transduction mechanisms in reproductive tissues. AB - Activins were identified initially as gonadal proteins having a stimulating effect on FSH production by the pituitary gland. Strong evidence has accumulated that activins are important regulating factors for many reproductive processes. Activin may have paracrine or autocrine roles rather than solely an endocrine action on FSH secretion. Activins together with their signalling molecules must be shown to be produced locally in a particular tissue to provide support for their paracrine or autocrine action in that tissue. The discovery of the activin receptors, the intracellular signalling mediators (Smads) and some transcription co-factors involved in activin responses has helped to unravel the activin transforming growth factor beta signalling mechanism. However, few reports have clearly demonstrated the presence of all of the activin signalling molecules in reproductive tissues, despite the important roles of activin in these tissues. Several activin receptor types and Smad molecules have been identified, indicating either a redundancy in signalling molecules or different signalling pathways. At present, it is not clear which particular subset of these signalling molecules is important in reproductive processes. The aim of this review is to collate the information available on activin actions, as well as on the signalling molecules, to understand how activins may transduce their paracrine or autocrine signals in reproductive tissues. PMID- 11427155 TI - Mechanisms for pattern formation leading to axis formation and lineage allocation in mammals: a marsupial perspective. AB - Developing patterns in early embryogenesis are analysed in conceptuses from several families, including Dasyuridae, Phalangeridae, Macropodidae and Didelphidae, in which cleavage has been examined in some detail. Features common to cleavage and blastocyst formation, and in some cases to hypoblast formation, are used to develop an outline of possible mechanisms leading to axis formation and lineage allocation. Relevant features that have been described only in some species are also included. It is suggested that certain features of marsupial cleavage establish patterns in the developing blastocyst epithelia, pluriblast, trophoblast and hypoblast that contribute to axis formation and lineage allocation. All marsupials examined had a polarized oocyte or conceptus, the polarity of which was related to the conceptus embryonic-abembryonic axis and, eventually, the conceptus dorsal-ventral axis and the formation of the pluriblast (future embryo) and trophoblast. The embryonic dorsal-ventral and anterior posterior axes were established after the allocation of hypoblast and epiblast. Mechanisms that appear to result in patterning of the developing epithelia leading to axis formation and lineage allocation are discussed, and include sperm entry point, gravity, conceptus polarity, differentials in cell-zona, cell-cell and cell-type (boundary effects) contacts, cell division order during cleavage and signals external to the conceptus. A model of the patterning effects is included. The applicability of these mechanisms to other amniotes, including eutherian mammals, is also examined. PMID- 11427156 TI - Mammalian aromatases. AB - Aromatase is the enzyme complex that catalyses the synthesis of oestrogens from androgens, and therefore it has unique potential to influence the physiological balance between the sex steroid hormones. Both aromatase cytochrome P450 (P450arom) and NADPH-cytochrome P450 reductase (reductase), the two essential components of the enzyme complex, are highly conserved among mammals and vertebrates. Aromatase expression occurs in the gonads and brain, and is essential for reproductive development and fertility. Of interest are the complex mechanisms involving alternative promoter utilization that have evolved to control tissue-specific expression in these tissues. In addition, in a number of species, including humans, expression of aromatase has a broader tissue distribution, including placenta, adipose and bone. The relevance of oestrogen synthesis and possibly androgen metabolism in these peripheral sites of expression is now becoming clear from studies in P450arom knockout (ArKO) mice and from genetic defects recognized recently in both men and women. Important species differences in the physiological roles of aromatase expression are also likely to emerge, despite the highly conserved nature of the enzyme system. The identification of functionally distinct, tissue-specific isozymes of P450arom in at least one mammal, pigs, and several species of fish indicates that there are additional subtle, but physiologically significant, species-specific roles for aromatase. Comparative studies of mammalian and other vertebrate aromatases will expand understanding of the role played by this ancient enzyme system in the evolution of reproduction and the adaptive influence of oestrogen synthesis on general health and well being. PMID- 11427157 TI - Variation in antral follicle development during the follicular phase of the oestrous cycle in red deer (Cervus elaphus) hinds. AB - The aim of this study was to quantify antral follicle populations in cyclic red deer hinds and to monitor follicle development leading to ovulation. Oestrus was synchronized with exogenous progesterone and ovaries were recovered approximately 0, 12, 24 or 36 h (follicular phase) or 10 days (luteal phase) after progesterone withdrawal (n = 5 per group). All follicles > or = 2 mm in diameter were dissected out, health status was assessed, follicular fluid oestradiol content was measured, granulosa cells were harvested and their capacity for oestradiol and cAMP production was determined. The time of oestrus and the preovulatory LH surge were monitored in five control hinds. Deer ovaries contained 26.6 +/- 3.45 (mean +/- SEM) follicles > or = 2 mm in diameter (range 4-81), with at least one large antral follicle (diameter: 8.3 +/- 0.38 mm) per hind. There was a strong correlation between follicle size and granulosa cell population (r(2) = 0.676). Approximately half (50.7%) of the follicles were classified as healthy, with the percentage classified as atretic decreasing with increasing follicle size. Neither the total number of antral follicles nor their size distribution differed significantly among groups. There were significantly more (P < 0.05) healthy follicles at 24 h after progesterone withdrawal than at 0 h, when large oestrogenic follicles had fewer granulosa cells, lower follicular fluid oestradiol concentrations and lower aromatase activity (P < 0.05) than did those from other groups. In summary, antral follicle development in red deer is similar to that in other monovulatory ruminants, and at least one large follicle is present at all stages of the oestrous cycle. PMID- 11427158 TI - Effect of immunization against the alpha N (alphaN) and alpha C (alphaC) peptides of the alpha43 subunit of inhibin on antral follicular growth and atresia and the patterns of gonadotrophin secretion in ewes. AB - The aims of this study were to investigate the role of inhibin in the distribution of healthy and atretic antral follicles and the secretion patterns of gonadotrophins. Ewes were actively immunized against either alphaN or alphaC of the inhibin alpha subunit with a primary injection and three booster injections. The control ewes received adjuvant only. The ovaries were removed either before or at 24 h after hCG administration in a synchronized follicular phase 48 h after removal of intravaginal progesterone pessaries. Morphological observations were made on every fifth section of the complete ovary (one per ewe) stained with haematoxylin and eosin. The mean number of corpora lutea observed per ewe with corpora lutea was not significantly different in ewes immunized against alphaN (2.4; alphaN-immunized ewes) or alphaC (2.6; alphaC-immunized ewes), and control (2.4) ewes, although some corpora lutea appeared cystic in the immunized ovaries. Compared with luteal phase concentrations, mean basal FSH concentrations in the early follicular phase were significantly increased in the alphaC-immunized ewes, similar in alphaN-immunized ewes and reduced in control ewes. No differences were observed in any of the LH parameters. Before hCG treatment, healthy antral follicles > 1 mm in diameter were not observed in any of the 52 follicles in the aC-immunized ewes and were observed in one of 37 follicles from alphaN-immunized ewes compared with 19 of 28 follicles in control ewes (P < 0.0001). For healthy antral follicles < 1 mm in diameter, there were 72 of 85 follicles in the alphaC-immunized ewes, 79 of 81 follicles in the alphaN immunized ewes and 81 of 82 follicles in the control ewes. Similar results were obtained in healthy antral follicles < 1 mm in diameter at 24 h after hCG administration. In contrast to the control ewes, no healthy preovulatory follicles (> 6 mm in diameter) were observed in alphaN- and alphaC-immunized ewes either before or 24 h after hCG administration. Two newly formed corpora lutea from alphaC-immunized ovaries contained retained oocytes compared with none in control and alphaN-immunized ovaries. In conclusion, immunization against alphaN and alphaC may result in disruption of the normal processes of antral follicular growth and maturation independent of the concentrations of FSH and LH. PMID- 11427159 TI - Possible role of kallikrein in proteolysis of insulin-like growth factor binding proteins during the oestrous cycle and early pregnancy in pigs. AB - During early pregnancy, pig conceptuses initiate the synthesis of oestrogens and on day 12 their trophoblastic membranes undergo a rapid expansion throughout the uterine horns. The insulin-like growth factor (IGF) system may be involved with conceptus development and steroidogenesis in pigs. Changes in uterine luminal IGF, insulin-like growth factor binding proteins (IGFBPs) and enzymatic activity for cleavage of IGFBPs during the oestrous cycle and early pregnancy were investigated. Uterine luminal content of IGF-I and IGF-II in uterine flushings from pigs on day 12 of pregnancy were two and three times greater, respectively, compared with uterine flushings collected from gilts during the oestrous cycle. Both IGF-I and -II content decreased on day 15 of gestation but content of IGF-II in uterine flushings remained three times greater than that of cyclic gilts. IGFBP-2 and -3 were the predominant binding proteins present in uterine flushings during days 0-10 of the oestrous cycle or day 10 of pregnancy. No IGFBPs were detected in the uterine flushings of either cyclic or pregnant pigs after day 10 by ligand blotting. Incubation of [125I]-labelled IGFBPs with various protease inhibitors indicated that cleavage of [125I]-labelled IGFBP-2 and -3 in uterine flushings involved serine proteases such as tissue kallikrein and matrix metalloproteinases. The results of the present study indicate that an increase in tissue kallikrein activity on day 12 of the oestrous cycle and pregnancy in pigs can directly, or indirectly through activation of matrix metalloproteinases, cleave IGFBP-2 and -3, thus allowing uterine release of IGF-I and -II in the uterine lumen to stimulate conceptus development. PMID- 11427160 TI - Full-term development of enucleated mouse oocytes fused with embryonic stem cells from different cell lines. AB - The developmental potential of enucleated mouse oocytes receiving embryonic stem cells from ten lines with either the same or different genetic backgrounds using the cell fusion method was examined in vitro and in vivo. The development of nuclear-transferred oocytes into blastocysts was high (34-88%). However, there was no clear correlation between development into blastocysts after nuclear transfer and the chimaera formation rate of embryonic stem cells. The development into live young was low (1-3%) in all cell lines and 14 of 19 young died shortly after birth. Most of the live young had morphological abnormalities. Of the five remaining mice, two died at days 23 and 30 after birth, but the other three mice are still active at days 359 (mouse 1) and 338 (mice 4 and 5) after birth, with normal fertility. However, the reasons for the abnormalities and postnatal death of embryonic stem cell-derived mice are unknown. PMID- 11427161 TI - Characterization of the active site of monkey sperm hyaluronidase. AB - The mammalian sperm hyaluronidase, PH-20, is active in macaque spermatozoa at neutral and acid pH. Antibodies were produced to synthesized peptides representing regions of PH-20 that may be involved in hyaluronidase activity and designated peptide 1 (amino acid sequence 142-172) and peptide 3 (amino acid sequence 277-297). Western blotting of proteins extracted from the surface of acrosome-intact spermatozoa showed that the two peptide-specific, affinity purified IgGs label a 64 kDa band corresponding to the PH-20 molecule. Western blots of acrosome-reacted spermatozoa showed that, under reducing conditions, the two anti-peptide IgGs label the 44 kDa band only, which represents the N-terminal fragment of PH-20. Anti-peptide 3 IgG also labels the 53 kDa form of PH-20 in extracts of acrosome-reacted spermatozoa. Peptide-specific, affinity-purified Fab fragments from both IgGs were shown by fluorescence microscopy and transmission electron microscopy to label the sperm plasma membrane, fused acrosomal vesicles, acrosomal matrix and inner acrosomal membrane. Fab fragments of anti-peptide 1 IgG, but not anti-peptide 3 IgG, inhibited hyaluronidase activity of PH-20 from the sperm surface and from extracts of acrosome-reacted spermatozoa at pH 7.0. Fab fragments of both anti-peptide IgGs inhibited sperm hyaluronidase activity at pH 5.0. It is concluded that the region of PH-20 encompassed by the amino acid sequence 142-172 is essential for hyaluronidase activity at neutral pH, whereas the region of amino acid sequence 277-297 may be more important at a lower pH. It is likely that these two regions are the acid/base catalyst site and the nucleophilic site, respectively, of PH-20 hyaluronidases. PMID- 11427162 TI - Delayed effect of heat stress on steroid production in medium-sized and preovulatory bovine follicles. AB - During the autumn, the conception rate of dairy cattle in warm countries is low although ambient temperatures have decreased and cows are no longer exposed to summer thermal stress, indicating that there may be a delayed effect of heat stress on cattle fertility. Two experiments were conducted to examine possible delayed effects of heat stress on follicular characteristics and steroid production at two distinct stages of follicular growth: medium-sized and preovulatory follicles, 20 and 26 days after heat exposure, respectively. Lactating cows were subjected to heat stress for 12 h a day in an environmental chamber, during days 2-6 of a synchronized oestrous cycle. In Expt 1, ovaries were collected on day 3 of the subsequent cycle, before selection of the dominant follicle, and medium-sized follicles were classified as atretic or healthy. In Expt 2, on day 7 of the subsequent cycle, PGF(2a) was administered and preovulatory follicles were collected 40 h later. In both experiments, follicular fluid was aspirated, granulosa and thecal cells were incubated, and steroid production was determined. In healthy medium-sized follicles (Expt 1), oestradiol production by granulosa cells and androstenedione production by thecal cells were lower (P < 0.05) and the concentration of progesterone in the follicular fluid was higher in cows that had been previously heat-stressed than in control cows (P < 0.05). In preovulatory follicles (Expt 2), the viability of granulosa cells was lower (P < 0.05) and the concentration of androstenedione in the follicular fluid and its production by thecal cells were lower (P < 0.05) in cows that had been previously heat-stressed than in control cows. In both experiments, the oestradiol concentrations in the follicular fluids were not altered by heat stress. These results demonstrate a delayed effect of heat stress on steroid production and follicular characteristics in both medium-sized and preovulatory follicles; this effect could be related to the low fertility of cattle in the autumn. PMID- 11427163 TI - Mechanisms of cytokine-induced death of cultured bovine luteal cells. AB - Tumour necrosis factor alpha (TNF-alpha) and gamma-interferon (IFN-gamma) are cytotoxic to bovine luteal cells in vitro and may contribute to cell death during luteolysis in vivo. In this study, the mechanism by which luteal cells are killed by TNF-alpha and IFN-gamma was investigated. Luteal cells were cultured for 7 days in the presence or absence of TNF-alpha and IFN-gamma. Inhibitors of arachidonate metabolism or scavengers of free radicals were included in the culture media. In addition, the effect of IFN-beta on the viability of cytokine treated luteal cells was tested. Lastly, untreated and cytokine-treated cells were subjected to single cell gel electrophoresis for quantification of DNA fragmentation. Neither indomethacin nor nordihydroguaiaretic acid, which are inhibitors of cyclooxygenase and lipoxygenase, respectively, were able to prevent cytokine-induced cell death. Similarly, both the phospholipase A(2) inhibitor arachidonyltrifluoromethyl ketone and the nitric oxide synthase inhibitor N(G) monomethyl-L-arginine, were largely without effect. In contrast, while vitamin C did not significantly affect viability, superoxide dismutase plus catalase increased viability of cytokine-treated cells (P < 0.05), and IFN-beta prevented cell death (P < 0.05). Finally, while control cells remained free of DNA damage, TNF-alpha plus IFN-gamma induced significant amounts of DNA damage by 48 h after initiation of treatment (P < 0.05). In conclusion, reactive oxygen species, but not arachidonate metabolism or nitric oxide, contribute to cytokine-induced luteal cell death in vitro, and the process of cell death may be via apoptosis. Furthermore, IFN-beta may confer protective effects against cytokine-induced cell death in bovine luteal cells. PMID- 11427164 TI - Measurement of serum and peritoneal fluid LH concentrations as a diagnostic tool for human endometriosis. AB - A rapid, sensitive enzymeimmunoassay for the measurement of LH concentrations in serum and peritoneal fluid samples of healthy women and women with endometriosis is reported. The ligand (LH) was captured by a readily available, widely used and well-characterized monoclonal antibody (mAb, 518B7) generated against the beta subunit of bovine LH. This mAb, although specific for LH, shows very little species specificity and detects LH by radioimmunoassay in humans. A polyclonal antiserum raised in rabbits against hCG was conjugated to horseradish peroxidase and was used as the second antibody signal. This anti-hCG antiserum crossreacts with LH. The enzymeimmunoassay uses the standard human LH (hLH) preparations (NIADDK-hLH-I-3, AFP-827OB) and results are based on the relative concentrations of LH in serum and peritoneal fluid. Total assay time was < 3 h. The range of the standard curve was 0.002-0.500 ng LH per well and the lowest concentration of hLH that could be distinguished from zero concentration was 0.15 +/- 0.02 ng ml(-1) serum and 0.058 +/- 0.021 ng ml(-1) peritoneal fluid. Clinical diagnostic parameters for the LH enzymeimmunoassay showed a sensitivity of 85.71%, specificity 92.50%, efficiency 88.54%, positive predictive value 94.11% and negative predictive value 82.22%. The study was retrospective. Serum LH concentrations of women with endometriosis were 13.67 +/- 7.21 ng ml(-1), whereas serum LH concentrations of women in the control group were 4.52 +/- 2.03 ng ml( 1). One-way ANOVA showed significant differences (P < 0.001) between women with endometriosis and control groups. Women in the control group had peritoneal fluid LH values of 5.65 +/- 2.43 ng ml(-1), whereas peritoneal fluid LH values of 64.06 +/- 16.44 ng ml(-1) were obtained in women with endometriosis (P < 0.001). A cycle-dependent pattern of serum and peritoneal fluid LH concentration was observed in women in the control group, which was not observed in the peritoneal fluid of the group with endometriosis. The application of this assay to serum or peritoneal fluid samples provides the attractive possibility that it could be included in the panel of markers used for diagnosis of endometriosis. PMID- 11427165 TI - Role of prostaglandins in intrauterine migration of the equine conceptus. AB - Between at least day 9 and day 16 after ovulation the spherical equine conceptus migrates continuously throughout the uterine lumen, propelled by peristaltic myometrial contractions. This unusually long period of intrauterine movement ensures that the conceptus delivers its anti-luteolytic signal to the entire endometrium to achieve luteostasis. The present experiment tested the hypothesis that prostaglandins stimulate the myometrial contractions that result in the migration of the conceptus. Serial ultrasonographic examinations of the uteri of eight mares performed during 2 h periods between day 10 and day 18 of gestation recorded the pattern of conceptus migration before and after treatment with the cyclo-oxygenase inhibitor flunixin meglumine. Conceptus mobility was high between day 10 and day 14 after ovulation (4.3 +/- 0.8, 4.7 +/- 0.8 and 4.3 +/- 0.9 changes of location per h on day 10, day 12 and day 14, respectively), but was reduced immediately and markedly by an i.v. injection of flunixin meglumine (3.8 +/- 1.5, 1.8 +/- 0.8 and 0.7 +/- 0.2 location changes per h), thereby implicating prostaglandins as the primary stimulus for the myometrial contractions that drive migration of the conceptus. PMID- 11427166 TI - Appearance and number of follicles and change in the concentration of serum FSH in female bovine fetuses. AB - The aim of this study was to determine the relationships between early follicular development, the time at which follicles appear in early stages of follicular development, and changes in the serum concentrations of FSH in the female bovine fetus. Thirty-five female bovine fetuses aged between 59 and 285 days, as estimated from the crown-rump length, were obtained from dams at an abattoir. Serum samples were separated from fetal blood obtained from the heart. Fetal ovaries were removed and weighed. The content of oestradiol in one of the fetal ovaries and the serum concentrations of FSH and oestradiol were determined using radioimmunoassay. Sections of the other ovary were examined histologically for the appearance and number of follicles. The follicles were divided into four stages: primordial, primary, secondary and early antral. The appearance of primordial, primary, secondary and early antral follicles was observed at day 74, day 91, day 120 and day 150, respectively. Serum concentrations of FSH in female bovine fetuses increased between day 120 and day 150 of gestation. Fetal serum concentrations of oestradiol increased from day 120. The number of early antral follicles increased from day 180 together with an increase in the fetal ovarian content of oestradiol. These findings indicate that, in the female bovine fetus as well as in adult cows, the number of follicles and stages of follicular development are associated with changes in the concentration of FSH. PMID- 11427167 TI - Disomy frequency estimated by multicolour fluorescence in situ hybridization, degree of nuclear maturity and teratozoospermia in human spermatozoa. AB - The aim of this study was to determine whether sperm morphology and nuclear maturity are associated with an increase in the prevalence of disomy in human spermatozoa. Semen samples were obtained from 60 patients selected at random from a population of men undergoing semen analysis as part of consultation for infertility. Semen analysis and aniline blue staining were carried out on each ejaculate to assess nuclear maturity. Disomy frequencies were determined using 15 18 dual colour and X-Y-8 multicolour fluorescence in situ hybridization on the four groups of samples with the five lowest and the five highest teratozoospermia values (groups 1 and 2), and the five lowest and the five highest percentages of aniline blue staining (groups 3 and 4). The prevalence of autosomal disomy was significantly higher in group 4 compared with group 3, but differences between groups 1 and 2 were not significant. No significant differences in the prevalence of gonosomal disomies or diploidies were observed among the groups. These results indicate a link between chromosomal meiotic segregation and the dynamic process of nucleoproteins during gametogenesis. The difference observed between the frequency of autosomal and gonosomal disomy using aniline blue staining was unexpected and may be due to the extended isolation of the gonosomes in the heterochromatic body. PMID- 11427168 TI - Different types of oestrous cycle in two closely related South American rodents (Cavia aperea and Galea musteloides) with different social and mating systems. AB - A comparative approach was used to investigate two closely related South American rodent species: the wild cavy (Cavia aperea) and the yellow-toothed cavy (Galea musteloides). These species of wild guinea-pig inhabit different habitats and show divergent social and mating systems. Cavia have a polygynous mating system, whereas in Galea promiscuous mating occurs. These observations correspond to functional variations in sperm characteristics, which might be interpreted as adaptations to different reproductive patterns in the females of both species. Twenty-two female C. aperea and G. musteloides (11 of each species) were investigated to elucidate differences in the characteristics of the oestrous cycles. The experimental design consisted of three stages, in which the degree of contact between the sexes was successively changed. During these stages, the reproductive cycle of the females was monitored by examining the condition of the vaginal closure membrane, analysing vaginal smears and determining serum concentrations of oestrogens and progesterone. The results revealed different types of oestrous cycle in the two species. Female C. aperea showed periodical cycles with spontaneously occurring oestrous periods, ovulation and corpus luteum activity. In contrast, in female G. musteloides, oestrus was exclusively induced by the presence of a male. However, after the induction of oestrus, ovulation and corpus luteum activity occurred spontaneously without stimuli from copulation. To date this type of oestrous cycle has not been described for any other mammalian species. Thus, these data indicate that differences in the reproductive patterns of the females may have shaped the evolution of different sperm characteristics in the males. PMID- 11427169 TI - Enhancement of aromatase activity by D-aspartic acid in the ovary of the lizard Podarcis s. sicula. AB - The present study investigated the role of D-aspartic acid (D-Asp) in ovarian steroidogenesis and its effect on aromatase activity in the lizard, Podarcis s. sicula. It was determined that D-Asp concentrations vary significantly during phases of the reproductive cycle: they vary inversely with testosterone concentrations and directly with oestradiol concentrations in the ovary and plasma. Experimental treatment showed that administration of D-Asp induces a decrease in testosterone and an increase in oestradiol, and that treatment with other amino acids (L-Asp, D-Glu and D-Ala) instead of D-Asp has no effects. Experiments in vitro confirmed these results. Furthermore, these experiments showed an increase in aromatase activity, as the addition of D-Asp either to fresh ovarian tissue homogenate or to acetonic powder of ovarian follicles induced a significant increase in the conversion of testosterone to oestradiol. Aromatase activity is four times greater in the presence of D-Asp than in its absence. However, almost equivalent values of the two K(m) values (both approximately 25 nmol l(-1)) indicate that aromatase has the same catalytic properties in both cases. PMID- 11427170 TI - Evidence for the innervation of sperm storage tubules in the oviduct of the turkey (Meleagris gallopavo). AB - The presence of neural tissue and smooth muscle elements in the vicinity of the oviductal sperm storage tubules at the uterovaginal junction was assessed by several modes of light microscopy. Isolated neurones and small ganglia were identified in the uterovaginal junction of the turkey oviduct. The nerve cell bodies were observed in the tunica mucosa by bright field microscopy. Immunoreactivity against neurofilament antibody and recombinant fragment C of the tetanus toxin reacted with nerve fibres and the nuclei of neurones. Fluorescence microscopy and confocal laser scanning microscopy revealed that nerve fibres continued from the base of the tunica mucosa into the plicae. Axons appeared to terminate on, or run immediately adjacent to, individual sperm storage tubules. Neither phalloidin reacting with F-actin nor the monoclonal antibody against alpha-smooth muscle actin detected smooth muscle fibres in the tissue encapsulating individual sperm storage tubules. In contrast, F-actin was strongly localized in the apical region of the epithelial cells of the sperm storage tubule and in smooth muscle elements in the tunica mucosa and tunica muscularis. These observations present the first evidence for the innervation of the sperm storage tubules. It is suggested that a previously unrecognized neural factor may function in oviductal sperm storage in, and release of spermatozoa from, the sperm storage tubules of hens. PMID- 11427171 TI - Embryo implantation during the short luteal phase of the corn mouse, Calomys musculinus, and the apparent lack of a lactational diapause in South American murid rodents. AB - As the corn mouse, Calomys musculinus, has a short luteal phase (2-3 days) that is not prolonged after copulation, it was hypothesized that (i) implantation would occur at the end of this phase, that is, earlier than it occurs in most murid species that have been studied, and (ii) a lactational embryonic diapause would not occur during the luteal phase. These hypotheses were tested in females that had copulated during postpartum oestrus and were either lactating or not lactating. Data were recorded from day 3 to day 5 of pregnancy (day 1 = day after coitus), at both 03:00-05:00 h and 17:00-19:00 h. Evidence of implantation in both non-lactating and lactating animals was apparent at 03:00-05:00 h on day 4 (endometrial 'blue reaction' in all cases and failure to recover free uterine embryos in some cases) and implantation swellings appeared within 24 h in both groups. In another experiment, the increase in duration of interbirth intervals in continuously mated females and their correlation with the number of suckling young were compared among C. musculinus, C. laucha, Akodon molinae (South American murid species) and Peromyscus maniculatus (a North American murid in which a lactational embryonic diapause has been shown). The results were indicative of a lactational embryonic diapause in the North American species, but not in the South American species. It was concluded that in C. musculinus (i) implantation occurs at the end of the spontaneous luteal phase, and (ii) that a lactational embryonic diapause does not occur: the absence of a lactational embryonic diapause may be common to other South American murid rodents. PMID- 11427172 TI - Genetics of Parkinsonism: a review. AB - Idiopathic Parkinson's disease (IPD), a progressive neurodegenerative disorder, is a common cause of disability. No current therapies modify disease progression. The pathological hallmarks are the presence of Lewy bodies and massive loss of dopaminergic neurons in the pars compacta of the substantia nigra. Two genes (SNCA and parkin) as well as two gene loci have now been implicated in the pathogenesis of familial PD. These represent significant progress in our understanding of the disease, considering the rarity of large families, low heritability in the general population and genetic heterogeneity. Mutations in a further gene, UCHL1, have been described in familial PD although the evidence for its role in PD is less clear. Knowledge of the genes described in PD to date should help to define molecular mechanisms of neurodegeneration in PD, as well as in other diseases where defects in protein handling may be a common feature. Nigral degeneration with Lewy body formation and the resulting clinical picture of PD may represent a final common pathway of a multifactorial disease process in which both environmental and genetic factors have a role. This review discusses the major advances in the field to date and illustrates how the existence of genetic factors has now become firmly established. PMID- 11427173 TI - Distribution of FMR1 and FMR2 alleles in Javanese individuals with developmental disability and confirmation of a specific AGG-interruption pattern in Asian populations. AB - The number of trinucleotide repeats in the 5' untranslated regions of the FMR1 and FMR2 genes was determined by PCR in 254 Fragile XA-negative Javanese male children with developmental disabilities. The distribution of FMR1 and FMR2 trinucleotide repeat alleles was found to be significantly different in the Indonesian population with developmental disability compared to that in developmentally disabled populations in North America and Europe (p & 0.021). Sequence analysis was performed on the trinucleotide repeat arrays of the 27 individuals with FMR1 alleles in the 'grey zone' (35-54 repeats). A repeat array structure of 9A9A6A9 was found in 16 unrelated individuals with 36 repeats, confirming earlier observations in intellectually normal Japanese. We propose that this FMR1 array pattern is specific for Asian populations and that Javanese and Japanese populations arose from a single progenitor population. PMID- 11427174 TI - MtDNA from extinct Tainos and the peopling of the Caribbean. AB - Tainos and Caribs were the inhabitants of the Caribbean when Columbus reached the Americas; both human groups became extinct soon after contact, decimated by the Spaniards and the diseases they brought. Samples belonging to pre-Columbian Taino Indians from the La Caleta site (Dominican Republic) have been analyzed, in order to ascertain the genetic affinities of these groups in relation to present-day Amerinds, and to reconstruct the genetic and demographic events that took place during the peopling of the Caribbean. Twenty-seven bone samples were extracted and analyzed for mtDNA variation. The four major Amerindian mtDNA lineages were screened through amplification of the specific marker regions and restriction enzymatic digestion, when needed. The HVRI of the control region was amplified with four sets of overlapping primers and sequenced in 19 of the samples. Both restriction enzyme and sequencing results suggest that only two (C and D) of the major mtDNA lineages were present in the sample: 18 individuals (75%) belonged to the C haplogroup, and 6 (25%) to the D haplogroup. Sequences display specific substitutions that are known to correlate with each haplogroup, a fact that helped to reject the possibility of European DNA contamination. A low rate of Taq misincorporations due to template damage was estimated from the cloning and sequencing of different PCR products of one of the samples. High frequencies of C and D haplogroups are more common in South American populations, a fact that points to that sub-continent as the homeland of the Taino ancestors, as previously suggested by linguistic and archaeological evidence. Sequence and haplogroup data show that the Tainos had a substantially reduced mtDNA diversity, which is indicative of an important founder effect during the colonization of the Caribbean Islands, assumed to have been a linear migratory movement from mainland South America following the chain configuration of the Antilles. PMID- 11427175 TI - Mitochondrial DNA sequence diversity in two groups of Italian Veneto speakers from Veneto. AB - Although frequencies of mitochondrial DNA (mtDNA) haplogroups in the different European populations are rather homogenous, there are a few European populations or linguistic isolates that show different mtDNA haplogroup distributions; examples are the Saami and Ladin speakers from the eastern Italian Alps. MtDNA sequence diversity was analysed from subjects from two villages in Veneto. The first, Posina, is situated in the Venetian Alps near Vicenza. The second, Barco di Pravisdomini is a village on the plains near Venice. In spite of their common Veneto dialect, the two group populations have not preserved a genetic homogeneity; particularly, they show differences in T and J haplogroups frequencies. MtDNA diversity in these two groups seems to depend more on their geographic situation. PMID- 11427176 TI - Mortality and cancer incidence in persons with Down's syndrome, their parents and siblings. AB - A cohort study of 1425 persons with Down's syndrome (DS), and of their parents (447 mothers, 435 fathers) and siblings (1176), was set up to investigate death rates from various causes and cancer incidence patterns. In individuals with DS the all-cause death rate was six times that of the national population (SMR = 622: 95% CI 559-693), the excess being attributable to many different causes. These included: leukaemia (SMR = 1304: 95% CI 651-2334); diabetes mellitus (SMR = 982: 95% CI 267-2515); Alzheimer's disease (SMR = 22028: 95% CI 7137-51326); epilepsy (SMR = 1727: 95% CI 744-3403); and congenital anomalies (SMR = 4987: 95% CI 4175-5955). The overall survival showed marked improvements for successive birth cohorts, particularly at young ages. For mothers and fathers of persons with DS, all-cause death rates were 20% lower than national rates and there were no significant excesses from any specific cause. For siblings, all-cause death rates were similar to national rates; the only condition with a significantly raised mortality ratio was colo-rectal cancer (SMR = 793: 95% CI 216-2031), but this may well be a chance finding. PMID- 11427177 TI - Mortality and cancer incidence in persons with numerical sex chromosome abnormalities: a cohort study. AB - Mortality and cancer incidence were assessed in a cohort of 1373 patients with numerical sex chromosome abnormalities diagnosed at three cytogenetics centres in Britain during 1959-90, and were compared with expectations from national rates. Four hundred patients with Turner's syndrome were followed, of whom 62 died, with a relative risk (RR) of death of 4.16 (95% confidence interval (CI) 3.22-5.39). Turner's syndrome patients had greatly raised risks of death from diseases of the nervous, cardiovascular, respiratory, digestive and genitourinary systems. One hundred and sixty three deaths occurred among 646 patients with Klinefelter's syndrome with a 47,XXY constitution, giving an RR of 1.63 (1.40-1.91). Mortality in these patients was significantly raised from diabetes and diseases of the cardiovascular, respiratory and digestive systems. There was also significantly increased mortality for patients with X polysomy (RR = 2.11 (1.43-3.02)) and Y polysomy (RR = 1.90 (1.20-2.85)), the former with significantly increased mortality from cardiovascular disease and the latter from respiratory disease. The only significantly raised risks of cancer incidence or mortality in the cohort were for lung cancer and breast cancer in patients with Klinefelter's syndrome with a 47,XXY constitution, and non-Hodgkin's lymphoma in men with more than three sex chromosomes. PMID- 11427178 TI - A permutation procedure for the haplotype method for identification of disease predisposing variants. AB - Once a genetic region involved in a complex disease has been localized through linkage or association studies, we need methods to help us identify the actual disease predisposing genetic variant(s) in the region. A large number of single nucleotide polymorphic (SNP) sites may exist in this region. It is important to identify genetic variants directly involved in disease from those in linkage disequilibrium, and thus associated with, the disease predisposing variant(s). A question of great interest is to test whether a SNP, or a combination of SNPs, that influence the trait under investigation have been identified. For many complex HLA-associated diseases, patterns of amino acid site variability raise the possibility that HLA-variation association with a disease may not be due to a given allele but rather one or more variable amino acid sites occurring on several alleles. Here the question is whether an amino acid variant or a combination of amino acid variants involved in disease are identified. To address this question, this paper proposes a permutation procedure for the haplotype method, to test whether all the sites involved in the disease have been identified using the haplotypic data of patients and controls. The method is based on the theoretical result of Valdes and Thomson, that, for each haplotype combination containing all the amino acid sites involved in the disease process, the relative frequencies of amino acid variants at sites not involved in disease, but in linkage disequilibrium with the disease-predisposing sites, are expected to be the same in patients and controls. This procedure takes into account the non-independence of the sites sampled and is robust to mode of inheritance and penetrance of the disease, and can definitely specify when all the disease predisposing sites have not been identified. Application to both simulated data and real data sets on type 1 diabetes and alcoholism indicates that the proposed procedure works well in testing the important null hypothesis of whether all the predisposing sites are identified. PMID- 11427179 TI - Fine-scale mapping using Hardy-Weinberg disequilibrium. AB - Hardy-Weinberg disequilibrium (HWD) among affected individuals has recently been proposed for fine-scale mapping of disease susceptibility genes. We investigate the statistical properties of several available HWD measures and develop a new HWD measure J for fine-scale mapping. It is shown both theoretically and through simulations that the available HWD measures depend not only on the genetic distance between the marker locus of interest and the disease susceptibility locus, but also on the allele frequencies at the marker locus. On the contrary, the new measure is not affected by the allele frequencies at the marker locus under the following assumptions: (a) there is initial complete linkage disequilibrium between the marker and the disease loci, (b) there are no new mutations at the marker and the disease loci, and (c) the population under study is large. We develop a novel method to estimate the location of the disease susceptibility gene based on the HWD measure J. The estimator is robust to low mutation rates at the marker and the disease loci. We compare the standard error of the estimated disease gene loci using P excess for case-control studies with the standard error using J for case-only studies under various disease models. The newly developed method is successfully applied to a data set on hereditary haemochromatosis (HH). PMID- 11427180 TI - Physical mapping: integrating computational and molecular genetic data. AB - A crucial step beyond the identification of genetic linkage of a disease to a chromosomal region is the production of a physical map that will allow the identification of candidate genes. Although the process of physical map building has been facilitated by the flow of data released by the Human Genome Project, gathering all the information together requires significant effort. In a previous study, we reported linkage between Bipolar Affective Disorder and the chromosomal location 4p15.3--p16.1. In this review we use this example to describe how to collect publicly available sequence, DNA fingerprint, and genetic marker data and integrate these with empirical data to build a large scale high resolution physical map of a region. Methods used to identify new genetic markers and candidate genes within a circumscribed region are also presented. PMID- 11427181 TI - Novel mutations in the 7-dehydrocholesterol reductase gene of 13 patients with Smith--Lemli--Opitz syndrome. AB - Smith--Lemli--Opitz syndrome (SLOS) is caused by mutations in the DHCR7 gene leading to deficient activity of 7-dehydrocholesterol reductase (DHCR7; EC 1.3.1.21), the final enzyme of the cholesterol biosynthetic pathway, resulting in low cholesterol and high concentrations of its direct precursor 7 dehydrocholesterol in plasma and tissues. We here report mutations identified in the DHCR7 gene of 13 children diagnosed with SLOS by clinical and biochemical criteria. We found a high frequency of the previously described IVS8--1 G > C splice acceptor site mutation (two homozygotes, eight compound heterozygotes). In addition, 13 missense mutations and one splice acceptor mutation were detected in eleven patients with a mild to moderate SLOS-phenotype. The mutations include three novel missense mutations (W182L, C183Y, F255L) and one novel splice acceptor site mutation (IVS8--1 G > T). Two patients, homozygous for the IVS8--1 G > C mutation, presented with a severe clinical phenotype and died shortly after birth. Seven patients with a mild to moderate SLOS-phenotype disclosed compound heterozygosity of the IVS8--1 G > C mutation in combination with different novel and known missense mutations. PMID- 11427182 TI - Interaction of the common apolipoprotein C-III (APOC3 -482C > T) and hepatic lipase (LIPC -514C > T) promoter variants affects glucose tolerance in young adults. European Atherosclerosis Research Study II (EARS-II). AB - Both hepatic lipase (HL) and apolipoprotein C-III (apoC-III) influence lipid metabolism. Common variation in promoters of both genes, LIPC -514C > T and APOC3 -482C > T, respectively, have been shown to affect plasma lipids and lipoproteins and glucose tolerance. We studied the interaction between both variants on parameters of glucose tolerance and lipid metabolism in 714 healthy young males participating in the second European Atherosclerosis Research Study (EARS-II). Approximately 18% of the subjects were carriers of at least one rare LIPC and APOC3 allele. These subjects exhibited, after fasting and oral fat loading, the highest values of triglyceride-rich lipoproteins, but there was no significant interactive effect on any lipid variable. However, interaction occurred on basal diastolic blood pressure (p =0.036) and, during oral glucose tolerance testing, on peak (p = 0.0065) and area under the curve for glucose (p =0.049), and insulin (p = 0.035). This resulted in the highest diastolic blood pressure and lowest glucose tolerance in carriers of at least one rare allele of both genes. Thus gene:gene interaction between LIPC and APOC3, even in these healthy young males, leads to changes in parameters that are typically characteristic of Syndrome-X. PMID- 11427183 TI - HLA molecular markers in Tuvinians: a population with both Oriental and Caucasoid characteristics. AB - HLA class I and class II alleles have been studied for the first time in the Turkish-speaking Tuvinian population, which lives in Russia, North of Mongolia and close to the Altai mountains. Comparisons have been done with about 11000 chromosomes from other worldwide populations, and extended haplotypes, genetic distances, neighbor joining dendrograms and correspondence analyses have been calculated. Tuvinians show an admixture of Mongoloid and Caucasoid characters, the latter probably coming from the ancient Kyrgyz background or, less feasibly, more recent Russian Caucasoid admixture. However, Siberian population traits are not found and thus Tuvinians are closer to Central Asian populations. Siberians are more related to Na-Dene and Eskimo American Indians; Amerindians (from nowadays Iberian--America) are not related to any other group, including Pacific Islanders, Siberians or other American Indians. The 'more than one wave' model for the peopling of the Americas is supported. PMID- 11427184 TI - Y chromosome microsatellite variation in three populations of Jerba Island (Tunisia). AB - Six Y-chromosome linked microsatellites were typed in a sample of 135 unrelated males representing three different ethnic groups: Arabs, Berbers and Blacks of Jerba Island (Tunisia). Analysis of variation at the six Y chromosome STRs showed significant differences in allele distributions between the Black group and the two other Islander groups. The Black group revealed the highest level of genetic diversity as compared to Arabs and Berbers, while the latter group was the most homogeneous. Allele frequencies obtained for the three islander groups analysed were compared to data available for some European, Mediterranean and African populations. Principal-coordinate analyses showed genetic differentiation between the three geographically closed groups of Jerba. The absence of the YAP insertion marker and the position of Arabs and Jerban Blacks near the European cluster would suggest their relative 'admixture' with European populations. PMID- 11427185 TI - Evidence for population sub-structuring in Sao Tome e Principe as inferred from Y chromosome STR analysis. AB - Seven Y-chromosome STR loci, DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392 and DYS393 have been analysed in population samples of Angolares, Forros and Tongas, three ethnic groups from the African archipelago of Sao Tome e Principe (Gulf of Guinea). Complete typings were obtained for 103 chromosomes, which belonged to 79 different haplotypes. The mean heterozygosity per locus in the overall Sao Tomean sample was 0.566, with the highest value found among Forros and the lowest among Angolares. Angolares also showed the lowest level of haplotype diversity. On average, the mean pairwise difference between two random haplotypes from Angolares, Forros and Tongas was 4.69, 6.74 and 6.23 repeats, respectively. The genetic distances were found to be statistically significant between Angolares and Forros or Tongas. In accordance, AMOVA revealed that the percentage of variation attributable to differences among groups was only significant when we distinguished between Angolares and non-Angolares. Globally, these results indicate that, with respect to the pool of male lineages of Sao Tome e Principe, some genetic sub-structuring does exist, basically determined by the Angolares ethnic group. PMID- 11427186 TI - Sample size calculations for classical association and TDT-type methods using family data. AB - Transmission Disequilibrium Test (TDT)-based methods have been advocated by several authors for testing that a marker-phenotype association is actually due to linkage and not to uncontrolled stratification. As a pre-requisite of TDT-type methods is the presence of an association between marker and phenotype, one may wish to first investigate the association using a classical association study, and then to check by a TDT approach whether this association is actually due to linkage. We propose an estimating equation (EE) procedure, to compute analytically the minimum sample size of sibship data required to detect the association between a marker and a quantitative phenotype, and that required to confirm it by two TDT methods. We show that, when the marker allele frequency is low or high, the number of informative sibs needed in TDT-type methods can be lower than the number required in an association analysis, and even more so when the familial clustering is strong. However, in all cases, the number of sibs that need to be sampled to get the appropriate number of informative sibs for analysis is always larger for TDT methods than for an association study. In a phenotype first strategy, this number may be critical when investigating costly phenotypes. PMID- 11427187 TI - LOD score exclusion analyses for candidate genes using random population samples. AB - While extensive analyses have been conducted to test for, no formal analyses have been conducted to test against, the importance of candidate genes with random population samples. We develop a LOD score approach for exclusion analyses of candidate genes with random population samples. Under this approach, specific genetic effects and inheritance models at candidate genes can be analysed and if a LOD score is < or = - 2.0, the locus can be excluded from having an effect larger than that specified. Computer simulations show that, with sample sizes often employed in association studies, this approach has high power to exclude a gene from having moderate genetic effects. In contrast to regular association analyses, population admixture will not affect the robustness of our analyses; in fact, it renders our analyses more conservative and thus any significant exclusion result is robust. Our exclusion analysis complements association analysis for candidate genes in random population samples and is parallel to the exclusion mapping analyses that may be conducted in linkage analyses with pedigrees or relative pairs. The usefulness of the approach is demonstrated by an application to test the importance of vitamin D receptor and estrogen receptor genes underlying the differential risk to osteoporotic fractures. PMID- 11427188 TI - How to develop cost-conscious guidelines. AB - BACKGROUND: Clinical guidelines, defined as 'systematically developed statements to assist both practitioner and patient decisions in specific circumstances', have become an increasingly familiar part of clinical care. Guidelines are viewed as useful tools for making care more consistent and efficient and for closing the gap between what clinicians do and what scientific evidence supports. Interest in clinical guidelines is international and has its origin in issues faced by most healthcare systems: rising healthcare costs; variations in service delivery with the presumption that at least some of this variation stems from inappropriate care; the intrinsic desire of healthcare professionals to offer, and patients to receive, the best care possible. Within the UK, there is ongoing interest in the development of guidelines and a fast-developing clinical-effectiveness agenda within which guidelines figure prominently. Over the last decade, the methods of developing guidelines have steadily improved, moving from solely consensus methods to methods that take explicit account of relevant evidence. However, UK guidelines have tended to focus on issues of effectiveness and have not explicitly considered broader issues, particularly cost. This report describes the methods developed to handle benefit, harm and cost concepts in clinical guidelines. It reports a series of case studies, each describing the development of a clinical guideline; each case study illustrates different issues in incorporating these different types of evidence. HEALTH ECONOMICS AND CLINICAL GUIDELINES: There has been no widely accepted successful way of incorporating economic considerations into guidelines. Unlike other areas of guideline development, there is little practical or theoretical experience to direct the incorporation of cost issues within clinical guidelines. However, the reasons for considering costs are clearly stated: "health interventions are not free, people are not infinitely rich, and the budgets of [health care] programmes are limited. For every dollar's worth of health care that is consumed, a dollar will be paid. While these payments can be laundered, disguised or hidden, they will not go away" (Eddy DM. A manual for assessing health practices and designing practice policies: the explicit approach. Philadelphia: American College of Physicians; 1992). Such opportunity costs are a universal phenomenon. In the USA it has been recommended that every set of clinical guidelines should include information on the cost implications of the alternative preventive, diagnostic, and management strategies for each clinical situation. The stated rationale was that this information would help potential users to evaluate better the potential consequences of different practices. However, it was acknowledged that "the reality is that this recommendation poses major methodological and practical challenges" (Institute of Medicine. Guidelines for clinical practice: from development to use. Washington: National Academy Press; 1992). METHODS OF DEVELOPING CLINICAL GUIDELINES: A guideline development process summarises the technical information about the value of treatments in a manner that makes them accessible and ready for use in clinical practice, alongside information on contextual issues. The requirement is that the presentation of costs and benefits of treatments is methodologically sound, robust and accessible. This report includes a summary of the current best practice in evidence-based guideline development, including recent methodological advances. The manner in which cost and cost-effectiveness concepts have been successfully incorporated into the guideline process is introduced. GUIDELINE DEVELOPMENT CASE STUDIES: The 'cost effectiveness' sections of 11 guidelines are reported to illustrate both the range of methods used and the nature of the recommendations reached by the guideline development groups when considering the profile of consequences of treatments including costs. These guidelines are broadly grouped as: (1) those using qualitative evidence summary methods; (2) those using quantitative evidence summary methods and addressing relatively narrow clinical questions; (3) those using quantitative evidence summary methods and addressing a broad clinical area; (4) a guideline based upon a decision analysis model. CONCLUSIONS: The focus of this project was to explore the methods of incorporating cost issues within clinical guidelines. However, the process of reviewing evidence in guideline development groups is becoming increasingly sophisticated, not only in considerations of cost but also in review techniques and group process. At the outset of the project it was unclear how narrowly or broadly the concept of 'cost' could be considered. (ABSTRACT TRUNCATED) PMID- 11427189 TI - Extended scope of nursing practice: a multicentre randomised controlled trial of appropriately trained nurses and pre-registration house officers in pre-operative assessment in elective general surgery. AB - OBJECTIVES: (1) To determine whether pre-operative assessment carried out by an appropriately trained nurse (ATN) is equivalent in quality to that carried out by a pre-registration house officer (PRHO). (2) To assess whether pre-assessments carried out by ATNs and PRHOs are equivalent in terms of cost. (3) To determine whether assessments carried out by ATNs are acceptable to patients. (4) To investigate the quality of communication between senior medical staff and ATNs. DESIGN: The study design was principally a prospective randomised equivalence trial but was accompanied by additional qualitative assessment of patient and staff perceptions, and an economic evaluation. SETTING: The study was carried out at four NHS hospitals, three of which were teaching hospitals, in three NHS Trusts in Southampton, Sheffield and Doncaster. SUBJECTS: All patients attending at one site for assessment prior to general anaesthetic for elective general, vascular, urological or breast surgery were potentially included in the study. Of 1907 patients who were randomised, 1874 completed the study with a full evaluation. INTERVENTIONS: The intervention consisted of a pre-operative assessment carried out by either an ATN or a PRHO. Of the patients who completed the study with a full evaluation, 926 patients were randomised to the PRHO arm of the trial and 948 to the ATN arm. Three ATNs took part in the study, one from each centre, together with a total of 87 PRHOs. MAIN OUTCOME MEASURES: Immediately following the initial assessment of a patient by a PRHO or an ATN, one of a number of clinical research fellows, all specialist registrars in anaesthetics, repeated the assessment and recorded it on a study form, together with a list of investigations required. The clinical research fellow then evaluated the competency of the initial assessor by comparing the quality of their assessment with their own. Any deficiencies in ordering of investigations and referral to other specialities were met in order to maximise patient care. Three areas of ATN and PRHO performance were judged separately, history taking, examination and ordering of tests, and each was graded into one of four categories, the most important of which was under-assessment, which would possibly have affected peri-operative management. In the case of ordering of tests, it was possible to have both over- and under-assessed a patient on different tests. RESULTS: The pre-operative assessments carried out by the ATNs were essentially equivalent to those performed by the PRHOs in terms of under assessment that might possibly have affected peri-operative management, although there was variation between the ATNs in terms of the quality of history taking. This may be related to the low number of patients seen at one study site. PRHOs ordered significantly more unnecessary tests than the ATNs. The substitution of ATNs for PRHOs was calculated to be cost neutral. The results of the qualitative assessment showed that the use of ATNs for pre-operative assessment was acceptable to patients; however, there was no evidence that communication between senior medical staff and those carrying out pre-operative assessments was improved by their introduction. CONCLUSIONS: This study demonstrated no reason to inhibit the development of fully nurse-led pre-operative assessment, provided that the nurses are appropriately trained and maintain sufficient workload to retain skills. CONCLUSIONS--IMPLICATIONS FOR THE HEALTH SERVICE: ATNs provide an acceptable and efficient alternative to PRHOs for the purposes of routine pre operative assessment. Consideration will have to be given, however, to the positions of these nurses within the surgical team, and also to their career structure. CONCLUSIONS--RECOMMENDATIONS FOR FUTURE RESEARCH: Further research is needed in the following areas: (1) the extent and type of training needed for nurses undertaking the pre-operative assessment role; (2) the use, costs and benefits of routine pre-operative testing. PMID- 11427190 TI - Protein translocation across membranes. AB - Cellular membranes act as semipermeable barriers to ions and macromolecules. Specialized mechanisms of transport of proteins across membranes have been developed during evolution. There are common mechanistic themes among protein translocation systems in bacteria and in eukaryotic cells. Here we review current understanding of mechanisms of protein transport across the bacterial plasma membrane as well as across several organelle membranes of yeast and mammalian cells. We consider a variety of organelles including the endoplasmic reticulum, outer and inner membranes of mitochondria, outer, inner, and thylakoid membranes of chloroplasts, peroxisomes, and lysosomes. Several common principles are evident: (a) multiple pathways of protein translocation across membranes exist, (b) molecular chaperones are required in the cytosol, inside the organelle, and often within the organelle membrane, (c) ATP and/or GTP hydrolysis is required, (d) a proton-motive force across the membrane is often required, and (e) protein translocation occurs through gated, aqueous channels. There are exceptions to each of these common principles indicating that our knowledge of how proteins translocate across membranes is not yet complete. PMID- 11427191 TI - Surface-associated serum proteins inhibit the uptake of phosphatidylserine and poly(ethylene glycol) liposomes by mouse macrophages. AB - Serum proteins, acting as opsonins, are believed to contribute significantly to liposome-macrophage cell association and thus regulate liposome uptake by cells of the mononuclear phagocytic system (MPS). We studied the effect of serum protein on binding and uptake of phosphatidylglycerol-, phosphatidylserine-, cardiolipin-, and N,N-dioleyl-N,N-dimethylammonium chloride- (DODAC) containing as well as poly(ethylene glycol)- (PEG) containing liposomes by mouse bone marrow macrophages in vitro. Consistent with the postulated surface-shielding properties of PEG, protein-free uptake of liposomes containing 5 mol% PEG and either 20 mol% anionic phosphatidylserine or 20 mol% cationic DODAC was equivalent to uptake of neutral liposomes. In contrast to previous reports indicating that protein adsorption to liposomes increases uptake by macrophages, the presence of bound serum protein did not increase the uptake of these liposomes by cultured macrophages. Rather, we found that pre-incubating liposomes with serum reduced the uptake of liposomes containing phosphatidylserine. Surprisingly, serum treatment of PEG-containing liposomes also significantly reduced liposome uptake by macrophages. It is postulated that, in the case of phosphatidylserine liposomes, the bound serum protein can provide a non-specific surface-shielding property that reduces the charge-mediated interactions between liposomes and bone marrow macrophage cells. In addition, incubation of PEG-bearing liposomes with serum can result in a change in the properties of the PEG, resulting in a surface that is better protected against interactions with cells. PMID- 11427192 TI - Transmembrane topography of plasma membrane constituents in mung bean (Vigna radiata L.) hypocotyl cells. I. Transmembrane distribution of phospholipids. AB - The transmembrane distribution of phospholipids (PLs) in the plasma membrane (PM) of mung bean (Vigna radiata L.) hypocotyl cells was investigated using annexin V fluorescein isothiocyanate, porcine pancreas phospholipase A(2), and (31)P nuclear magnetic resonance (NMR) spectroscopy. Phosphatidylserine was not located on the cell surface of mung bean protoplasts. However, phosphatidylcholine, phosphatidylethanolamine and phosphatidic acid were found to be almost symmetrically distributed across right-side-out PM vesicles obtained by aqueous two-phase partitioning by porcine pancreas phospholipase A(2) assay. (31)P-NMR assay showed that the amount of PLs is about equal in the outer and the inner leaflets of the right-side-out PM vesicles. These results suggest that the topography of PM PLs might not contribute to well-known asymmetrical properties of the outer and inner surfaces of higher plant PMs. It is also indicated that inside-out PM vesicles created by Brij 58-treatment do not retain the native PL topography on dithionate reduction of 7-nitro-2,1,3-benzoxadiazol-4-yl-labeled PLs incorporated in the PM vesicles. PMID- 11427193 TI - Localization of the murine reduced folate carrier as assessed by immunohistochemical analysis. AB - The reduced folate carrier (RFC1) is a major route for the transport of folates in mammalian cells. The localization of RFC1 in murine tissues was evaluated by immunohistochemical analysis using a polyclonal antibody to the C-terminus of the carrier. There was expression of RFC1 in the brush-border membrane of the jejunum, ileum, duodenum and colon. RFC1 was localized to the basolateral membrane of the renal tubular epithelium. Carrier was detected on the plasma membrane of hepatocytes but not in bile duct epithelial cells. In the choroid plexus RFC1 was highly expressed at the apical surface. It was also expressed in axons and dendrites and on the apical membrane of cells lining the spinal canal. In spleen, RFC1 was detected only in the cells of the red pulp. These data provide insights into the role that RFC1 plays in folate delivery in a variety of tissues. In particular, the localization of carrier may elucidate the role of RFC1 in the vectorial transport of folates across epithelia. The data also indicate that in kidney tubules and choroid plexus the sites of RFC1 expression are different from what has been reported previously for the folate receptor; and while RFC1 is expressed in small intestine, folate receptor is not. PMID- 11427194 TI - Imaging mixed lipid monolayers by dynamic atomic force microscopy. AB - Phase imaging with tapping mode atomic force microscopy (AFM) and force modulation microscopy were used to probe the mechanical properties of phase separated lipid monolayers made of a mixture (0.25:0.75) of the surface-active lipopeptide surfactin and of dipalmitoylphosphatidylcholine (DPPC). The pi-A isotherms and the result of a molecular modeling study revealed a loose, 2-D liquid-like organization for the surfactin molecules and a closely packed, 2-D solid-like organization for DPPC molecules. This difference in molecular organization was responsible for a significant contrast in height, tapping mode phase and force modulation amplitude images. Phase imaging at light tapping, i.e., with a ratio of the set-point tapping amplitude with respect to the free amplitude A(sp)/A(0) approximately 0.9, showed larger phase shifts on the solid like DPPC domains attributed to larger Young's modulus. However, contrast inversion was observed for A(sp)/A(0)<0.7, suggesting that at moderate and hard tapping the image contrast was dominated by the probe-sample contact area. Surprisingly, force modulation amplitude images showed larger stiffness for the liquid-like surfactin domains, suggesting that the contrast was dominated by contact area effects rather than by Young's modulus. These data emphasize the complex nature of the contrast mechanisms of dynamic AFM images recorded on mixed lipid monolayers. PMID- 11427195 TI - Molecular dynamics simulations of the insertion of two ideally amphipathic lytic peptides LK(15) and LK(9) in a 1,2-dimyristoylphosphatidylcholine monolayer. AB - We present here the results of 1-ns molecular dynamics (MD) simulations of two ideally amphipathic lytic peptides, namely LK(15) and LK(9), in a 1,2 dimyristoylphosphatidylcholine monolayer with two different cross-sectional areas per lipid of 80 A(2) (loose film) and 63 A(2) (tight standard film). These peptides are lytic, ideally amphipathic with a minimalist composition L(i)K(j) and the following sequences: H(2)N-KLLKLLLKLLLKLLK-CO-Ph for LK(15) and H(2)N KLKLKLKLK-CO-Ph for LK(9). From experimental data, LK(15) exhibits an alpha helical secondary structure, whereas LK(9) was found to form antiparallel beta sheets at the interface of a DMPC monolayer. Whatever the specific lipid surface is, the two peptides exhibit very different behavior: the alpha-helix inserts deeply into the monolayer whereas the beta-sheeted peptide stays at the surface within the upper polar part of the monolayer. In all cases, a loose monolayer (80 A(2)) results in noticeable artifacts whereas a monolayer with standard specific surface leads to very reliable behavior well in accordance with experimental data. Despite their different insertion depth, the two peptides exhibit identical lytic efficiency. This is very likely a direct consequence of the very strong Van der Waals interactions between the fatty alkyl chains of the lipids and the highly lipophilic lower part of the peptide, resulting in an identical thinning of the two monolayers. PMID- 11427196 TI - Formation of liposomes by resorcinolic lipids, single-chain phenolic amphiphiles from Anacardium occidentale L. AB - Resorcinolic lipids isolated from Anacardium occidentale nut oil extract (CNSL), unsaturated congeners of those isolated from bacterial and graminaceous sources, form at alkaline conditions liposomal structures alone as well as in the mixtures with cholesterol, fatty acids or phosphatidylethanolamine. Those vesicular structures show relatively high entrapment of the marker and stability of their size. The retention of the captured solute depends upon the type of resorcinolic lipid and on the temperature, but in general, is lower than control phospholipid liposomes. PMID- 11427198 TI - The apoAI-CIII-AIV gene cluster. AB - The relationship between variations in the apoAI-CIII-AIV gene cluster and plasma lipid traits has been recognized for at least 15 years. Most studies have been focused upon the association between plasma triglycerides and a genetic variation present in the 3' untranslated region of the apoC-III gene (SstI polymorphism). Although not all studies showed similar results, this variant is most consistently found in association with plasma triglycerides. The other most interesting DNA variant is the mutation present in the promoter region of the apo A-I gene at position -75 (Msp-AI polymorphism), which has been associated with differences in plasma apoA-I and HDL-cholesterol levels. In this review, we will discuss the most frequent occurring genetic variants in the apoAI-CIII-AIV gene cluster and their impact on plasma lipid traits and associations with increased risk on developing coronary artery disease. PMID- 11427199 TI - Effects of the heparin-mimicking compound RG-13577 on lipoprotein lipase and on lipase mediated binding of LDL to cells. AB - Lipoprotein lipase (LPL) has high affinity for heparin and heparin-like compounds. In vivo the enzyme is attached to heparan sulfate proteoglycans on the endothelium of capillaries and larger blood vessels. The enzyme is released from these sites after intravenous injection of heparin. One has here investigated the effects of RG-13577 on LPL, both after intravenous injection to rats and under cell culture conditions. RG-13577 is a heparin-mimicking compound known to prevent angiogenesis by interference with binding of growth factors to cells. It has therefore been considered for use in cancer therapy as well as for prevention of atherosclerosis and restenosis. It was found that intravenously injected RG 13577 released both LPL and hepatic lipase (HL) to the blood. Binding of LPL in extrahepatic tissues was prevented and clearance of radiolabeled LPL from the circulation was delayed. Furthermore, RG-13577 released LPL from extracellular matrix (ECM) produced by endothelial cells and from THP-1 monocyte-derived macrophages. Lipase-mediated binding and uptake of human LDL in these cells was also prevented by RG-13577. Thus, in the test systems RG-13577 had the same effects as heparin, but on a molar basis RG-13577 was in all cases less effective. PMID- 11427200 TI - Time sequence of the inhibition of endothelial adhesion molecule expression by reconstituted high density lipoproteins. AB - We have used discoidal reconstituted high density lipoproteins (rHDL) containing apolipoprotein (apo) A-I and dimyristoyl phosphatidylcholine (DMPC) as a tool to investigate the time sequence of the HDL-mediated inhibition of vascular cell adhesion molecule (VCAM)-1 and E-selectin expression in cytokine-activated human umbilical vein endothelial cells (HUVECs). Specifically, we have asked a few questions - (i) how long do the cells need to be exposed to the rHDL before adhesion molecule expression is inhibited and (ii) how long does the inhibition persist after removing the rHDL from the cells. When the cells were not pre incubated with the rHDL, there was no inhibition. The magnitude of the inhibition increased progressively with increasing duration of pre-incubation up to 16 h. Inhibition did not require the rHDL to be physically present during the activation of adhesion molecule expression by tumour necrosis factor(TNF)-alpha, excluding the possibility that the rHDL was merely interfering with the interaction between TNF-alpha and the cells. When HUVECs were pre-incubated for 16 h with rHDL, the inhibition remained substantial even if the rHDL were removed from the medium up to 8 h prior to addition of TNF-alpha. The HDL-mediated inhibition of VCAM-1 in HUVECs was unaffected by the presence of puromycin, an inhibitor of protein synthesis, excluding the possibility that HDL may have acted by stimulating the synthesis of a cell protein that itself inhibits adhesion molecule expression. These results have important implications in terms of understanding the mechanism(s) of the HDL-mediated inhibition of endothelial adhesion molecule expression. PMID- 11427201 TI - Effect of dietary elaidic versus vaccenic acid on blood and liver lipids in the hamster. AB - Male hamsters (30 per group) were fed five different semi-purified diets ad libitum. The diets, containing 30% of energy (en%) as fat, differed in their dietary fat composition (specified fatty acids exchanged at 10 en%) and were fed for 4 weeks. The five fatty acids compared in mixed triglycerides were elaidic acid (C18:1 9t), vaccenic acid (C18:1 11t), their cis-counterpart oleic acid (C18:1 9c), medium-chain fatty acids (MCFA; C8:0 and C10:0), and palmitic acid (C16:0). Compared with oleic acid, dietary MCFA and palmitic acid tended to increase blood cholesterol levels in the hamsters. The effect of elaidic and vaccenic acid on blood cholesterol did not differ from that of oleic acid. When elaidic acid and vaccenic acids were compared directly, the ratio of LDL/HDL cholesterol in plasma was significantly higher in hamsters fed vaccenic acid than in those fed elaidic acid, and elaidic acid was incorporated at low levels, but more efficiently than vaccenic acid at the sn-2 position of platelet phospholipids. Biological consequences of this low incorporation are considered unlikely as levels of arachidonic acid (C20:4 n-6) and docosohexaenoic acid (C22:6 n-3) in the platelet phospholipids of all dietary groups did not differ. With respect to the effect on the LDL/HDL-cholesterol ratio, elaidic acid may be preferable to vaccenic acid. We conclude that this animal study does not provide evidence for the suggestion, based on epidemiological observations, that elaidic acid would be more detrimental to cardiovascular risk than vaccenic acid. PMID- 11427202 TI - Maternal atherogenic diet in swine is protective against early atherosclerosis development in offspring consuming an atherogenic diet post-natally. AB - Atherosclerosis found early in life suggests that it may arise from fetal adaptations during development in utero. We evaluated the effect of a maternal atherogenic diet on atherosclerosis development in offspring. Aortic fat deposition was evaluated by Sudan IV staining and coronary atherosclerosis was assessed histologically. Sows were fed an atherogenic diet (ath) or standard diet (std) throughout gestation. Twelve neonates (six per maternal diet group) were evaluated with no significant differences noted (P>0.05) in serum lipids or aortic fat deposition and there was no evidence of coronary atherosclerosis. Twenty offspring (10 per maternal diet group) were followed for 5 months forming the pubertal age group. Half of these swine received an atherogenic (ATH) diet (std-ATH and ath-ATH) and half received a standard (STD) diet (std-STD and ath STD). Pubertal age swine on the ATH diet had significantly greater (P<0.05) serum lipids and aortic fat deposition compared with those on the STD diet, with significantly greater fat deposition (P<0.05) occurring in the std-ATH versus ath ATH group. Coronary atherosclerosis was exhibited only in the std-ATH diet group. Our findings suggest that gestational diet may alter the body's management of cholesterol later in life, possibly providing a protective effect from atherosclerosis. PMID- 11427203 TI - Twenty four hour insulin infusion impairs the ability of plasma from healthy subjects and Type 2 diabetic patients to promote cellular cholesterol efflux. AB - Removal of cholesterol from peripheral cells by high density lipoproteins (HDL) is regarded as an important defence mechanism against atherosclerosis development. PLTP is involved in the generation of pre beta-HDL that can act as initial acceptors of cellular cholesterol. Exogenous hyperinsulinaemia may not only decrease HDL cholesterol, but also plasma phospholipid transfer protein (PLTP) activity. The effect of 24-h insulin infusion (30 mU/kg/h) on the ability of plasma to promote cholesterol efflux from Fu5AH cells was examined in eight healthy men and eight male Type 2 diabetic patients, matched for HDL cholesterol. Baseline HDL cholesterol and phospholipids, pre beta-HDL in incubated plasma, plasma apolipoprotein (apo) AI, PLTP activity and cholesterol efflux to plasma were not different between the groups. In both groups, HDL lipids, as well as plasma apo AI and PLTP activity decreased after 24 h of insulin (P<0.05 to P<0.01) compared to baseline and recovery, i.e. 1 week after insulin. Pre beta HDL in incubated plasma did not significantly change. Cholesterol efflux to plasma from both groups decreased after insulin (P<0.05). Using plasma from healthy subjects, cholesterol efflux was correlated positively with HDL cholesterol, HDL phospholipids, pre beta-HDL in incubated plasma, plasma apo AI and PLTP activity (P<0.05 to P<0.001). Using plasma from diabetic patients, cholesterol efflux was not significantly correlated with any of these parameters. In conclusion, 24-h moderate hyperinsulinaemia impairs the ability of plasma to promote cholesterol efflux from Fu5AH cells. It is suggested that, apart from HDL, plasma PLTP activity is a determinant of cholesterol efflux via stimulation of pre beta-HDL formation. Cellular cholesterol efflux to plasma from selected Type 2 diabetic patients is maintained, but the interaction of Fu5AH cells with HDL may be altered. PMID- 11427204 TI - Association of angiotensin converting enzyme and plasminogen activator inhibitor 1 promoter gene polymorphisms with features of the insulin resistance syndrome in patients with premature coronary heart disease. AB - Polymorphisms of the angiotensin-converting enzyme (ACE) (insertion/deletion (I/D) in intron 16) and of the plasminogen activator inhibitor-1 (PAI-1) (promoter 4G/5G) genes have been linked with coronary heart disease (CHD) and/or myocardial infarction (MI). We studied the association of polymorphisms in these genes with CHD with linkage and association analyses in 118 families with premature and severe CHD and in 110 healthy controls. In linkage analysis there was no evidence for a linkage of the ACE or PAI-1 loci with CHD. However, in quantitative linkage analysis the ACE locus was linked with fasting glucose (P=0. 047) and fasting free fatty acid levels (P=0.029). In association analysis the ACE genotype frequencies of probands with CHD did not differ from those of healthy controls. Normoglycemic probands with MI and with the ACE polymorphism DD genotype had characteristics of the insulin resistance syndrome. They had higher levels of 1-h glucose (P=0.008) and 2-h free fatty acids (P=0.011) in an oral glucose tolerance test and higher levels of total (P=0.005) and very-low-density lipoprotein triglycerides (P=0.006) than probands with the ID or the II genotypes. The PAI-1 gene polymorphism was not associated with any of the variables of glucose or lipid metabolism. In conclusion, the ACE and PAI-1 gene polymorphisms are not linked with early-onset CHD. However, the ACE gene polymorphism is associated with features of the insulin resistance syndrome. PMID- 11427205 TI - Reversible and irreversible non-internalized LDL and methyl LDL accumulation by human fibroblasts. AB - In previous in vivo animal studies, we showed that low density lipoprotein (LDL) accumulated irreversibly at the edges of healing arterial lesions rather than being internalized and degraded. To see if similar LDL accumulation occurs in vitro, fibroblasts from normal and homozygous familial hypercholesterolemic (FH) subjects were incubated at 37 degrees C with 125I-LDL and 125I-methyl LDL; the latter is not recognized by any known LDL receptor. Normal fibroblast accumulation of LDL and methyl LDL (5 microg/ml) plateaued within 1 h at 200 and 100 ng/mg, respectively. With FH cells, both LDL and methyl LDL accumulation plateaued at 100 ng/mg. Lipoprotein accumulation by both cell types rose steeply at concentrations up to 15-25 microg/ml, and less so at higher concentrations. Except for degradation of LDL by normal cells, degradation was minimal, which indicated that much of the lipoprotein accumulation was unaccompanied by internalization. The accumulation of both lipoproteins by both cell types was greater at 37 degrees C than at 4 degrees C, and was inhibited between 43 and 75% by homologous unlabeled lipoprotein. To see if any accumulation was irreversible, cells were incubated with radiolabeled lipoproteins for 3 h (pulse), then with homologous unlabeled lipoproteins for up to 20 h (chase). About 50% of intact radiolabeled lipoprotein rapidly dissociated from cells into the medium in the first 4 h of the chase period. In contrast, between 4 and 20 h, most of the remaining intact LDL and methyl LDL appeared to be irreversibly bound, because it was released at a rate of only 0-1%/h. Thus, we conclude that, under the conditions studied, both reversible and irreversible non-internalized LDL binding play a major role in LDL accumulation by cultured cells. PMID- 11427206 TI - Fibromuscular cap composition is important for the stability of established atherosclerotic plaques in mature WHHL rabbits treated with statins. AB - We examined the relationship between plaque vulnerability and fibromuscular cap composition using hydrophilic pravastatin and lipophilic fluvastatin. WHHL rabbits aged 10 months were given pravastatin (50 mg/kg) or fluvastatin (20 mg/kg) for 52 weeks. The atherosclerotic lesions were immunohistochemically or conventionally stained and the components were analyzed with a color image analyzer. Compared with the control group, the plasma cholesterol levels were decreased by about 25% in both statin groups. Pravastatin decreased the lipid components (macrophages+extracellular lipids) in whole aortic plaques by 34% and the fibrous caps of coronary plaques by 55%. Fluvastatin decreased the fibromuscular components (smooth muscle cells+collagen fibers) in whole aortic plaques and in the fibromuscular caps of the aortic and coronary plaques. In the pravastatin group, the vulnerability index, the ratio of (lipid components)/(fibromuscular components), was decreased in whole aortic plaques by 28% and in the fibromuscular caps of coronary lesions by 61%, while the indexes were increased in the fluvastatin group. The incidence of vulnerable plaques was decreased by 74% in the coronary plaques of the pravastatin group. Our results suggest that the stability of atheromatous plaques was improved due to a decrease of the lipid components and vulnerability index of the fibromuscular cap by pravastatin. PMID- 11427207 TI - Circulating lipoproteins and hepatic sterol metabolism in Psammomys obesus prone to obesity, hyperglycemia and hyperinsulinemia. AB - The liver plays a central role in lipoprotein metabolism and cholesterol homeostasis. As the physiopathology of lipid disorders in non-insulin-dependent diabetes mellitus (NIDDM) is multifactorial and still imperfectly known, we evaluated its onset on plasma lipid transport and hepatic cholesterol metabolism in Psammomys obesus. This sand rat lapses into hyperinsulinemia and hyperglycemia when transferred from its native food to laboratory rodent diets. Marked hypertriglyceridemia and hypercholesterolemia developed in hyperinsulinemic (Group B) and hyperglycemic/ hyperinsulinemic (Group C), compared with normal P. obesus (Group A). Group B showed significantly (P<0.05) higher plasma VLDL cholesterol (41.9%) and LDL-cholesterol (47.3%) concentrations, whereas Group C was characterized by an even more marked increase in VLDL-cholesterol (176%, P<0.001) compared with Group A. Lipoprotein composition was also altered, displaying impaired lipid and apolipoprotein moiety distribution in IDL, LDL, HDL(2) and HDL(3) lipoprotein fractions of Groups B and C. The activity of hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase, the rate-limiting enzyme in cholesterol biosynthesis, was consistently lower in Group B (P<63.4%, P<0.001) and C (43.9%, P<0.005). In contrast, the direct measurement of microsomal acyl CoA:cholesterol acyltransferase (ACAT), controlling the acylation of cholesterol, showed an increase averaging 53% in Group B (P<0.01) and 61% in Group C (P<0.005). Similarly, elevated activity (171.1%, P<0.05 and 291.4%, P<0.001, respectively) was related to cholesterol 7alpha-hydroxylase, the rate-limiting enzyme in bile acid biosynthesis. These alterations were accompanied with abundant deposition of triglycerides and cholesterol in the liver. Changes in circulating lipids and liver parameters were related to glucose and insulin levels, indicating the implication of insulin resistance and diabetes. Therefore, our findings demonstrate various disturbances in plasma lipid profile and lipoprotein composition, as well as in liver cholesterol metabolism during the sequential development of insulin resistance and diabetes in P. obesus rats. Furthermore, the current data point to an undoubtedly important role of the liver in the pathogenesis of metabolic disorders in the progression of nutritionally induced insulin resistance and diabetes in P. obesus. Finally, current research shows that more marked plasma and hepatic lipid perturbations occur in insulin resistance than in diabetes, which may culminate in the development of atherosclerosis. PMID- 11427208 TI - The combined effect of inhibiting both ACAT and HMG-CoA reductase may directly induce atherosclerotic lesion regression. AB - We hypothesized that coadministration of avasimibe and simvastatin would limit size, composition and extent of atherosclerotic lesions and potentially promote lesion regression, since bioavailable ACAT inhibitors decrease monocyte macrophage enrichment and HMG-CoA reductase inhibitors limit smooth muscle cell migration and proliferation. Male New Zealand white rabbits were sequentially fed a 0.5% cholesterol, 3% peanut oil, 3% coconut oil diet for 9 weeks and a chow-fat diet for 6 weeks prior to drug administration. A time zero control group was necropsied prior to drug administration and the progression control was fed various diets but untreated. Avasimibe (10 mg/kg), simvastatin (2.5 mg/kg) or combination of avasimibe (10 mg/kg) with simvastatin (2.5 mg/kg) were administered in the chow-fat diet for 8 weeks. Plasma total cholesterol exposure was unchanged by avasimibe but was reduced 21% by both simvastatin alone and in combination with avasimibe. Combination of avasimibe and simvastatin decreased VLDL-cholesterol exposure by 56%. VLDL+IDL lipid composition was similar in the progression control and simvastatin-treated animals. Administration of avasimibe alone or in combination with simvastatin reduced the cholesteryl ester fraction and increased the triglyceride fraction to comparable extents. Relative to the progression control, avasimibe plus simvastatin markedly decreased thoracic aortic cholesteryl ester content and lesion coverage by 50% and aortic arch lesion size and macrophage area by 75 and 73%, respectively. With respect to lesion regression, avasimibe+simvastatin decreased aortic arch lesion size by 64% and monocyte-macrophage area by 73% when compared to time zero. Based on these data, we conclude that despite changes in plasma total and lipoprotein cholesterol exposure and lipoprotein composition comparable to monotherapy, inhibition of both ACAT and HMG-CoA reductase may not only directly blunt lesion progression but also promote regression of pre-established atherosclerotic lesions. PMID- 11427209 TI - Prolonged inhibition of cholesterol synthesis by atorvastatin inhibits apo B-100 and triglyceride secretion from HepG2 cells. AB - Atorvastatin is a new HMG-CoA reductase inhibitor that strongly lowers plasma cholesterol and triglyceride (TG) levels in humans and animals. Since previous data indicated that atorvastatin has prolonged inhibition of hepatic cholesterol synthesis, we tested whether this longer duration of inhibitory effect on cholesterol synthesis decreased hepatic lipoprotein secretion in vitro. We used the HepG2 hepatoma cell line to: (1) determine the time required until levels of secreted apo B-100 and TG declined significantly, (2) examine the relation to the mass of cellular cholesteryl ester (CE) and (3) test microsomal triglyceride transfer protein (MTP) activity which leads to decreased apo B-100 production. Although atorvastatin significantly inhibited cholesterol synthesis in HepG2 cells regardless of treatment duration (1, 14 or 24 h), it did not inhibit TG synthesis. Apo B-100 and TG secretion were unchanged after 1-h atorvastatin treatment, but declined significantly after 24-h treatment. Atorvastatin treatment also reduced cellular CE mass, exhibiting both time- and dose dependency. Mevalonolactone, a product of HMG-CoA reductase, attenuated the inhibitory effects of atorvastatin. Atorvastatin strongly reduced mRNA levels of MTP, whereas it did not inhibit MTP activity as measured by TG transfer assay between liposomes. Simvastatin also induced treatment- and time-dependent reductions in apo B-100, whereas the MTP inhibitor BMS-201038 exhibited no time dependency, instead inhibiting this variable even on 1-h treatment. These results indicate that reduced apo B-100 secretion caused by atorvastatin is a secondary result owing to decreased lipid availability, and that atorvastatin's efficacy depends on the duration of cholesterol synthesis inhibition in the liver. PMID- 11427210 TI - The adventitia of atherosclerotic coronary arteries frequently contains Chlamydia pneumoniae. AB - The presence of Chlamydia pneumoniae in the human arterial system has mainly been determined in atherosclerotic plaque, whereas the adventitia has remained relatively unexplored. We assessed the presence of C. pneumoniae in all three vessel wall layers of coronary (n=72) and brachial (n=48) arteries in relation to local atherosclerosis. Immunohistochemical staining of C. pneumoniae was observed in plaque and adventitia. Cells stained for C. pneumoniae were detected in the same areas as cells stained for macrophages in adjacent sections. C. pneumoniae staining in the adventitia was associated with the extent and severity of atherosclerosis. Coronary sections with C. pneumoniae staining in both adventitia and plaque more often contained advanced atherosclerosis than sections with staining only in the adventitia. Staining was observed more often in the coronary artery than in the brachial artery (24/72 vs. 5/48 and 51/72 vs. 8/48 for plaque and adventitia, respectively, P=0.004 and P<0.001). PCR confirmed the presence of C. pneumoniae DNA in the adventitia. In summary, the adventitia of atherosclerotic coronary arteries frequently contains C. pneumoniae that seems to be located within macrophages. These results might indicate a possible route for infected circulating macrophages to home into atherosclerotic lesions in the artery via vasa vasorum. PMID- 11427211 TI - Lipoprotein lipase D9N, N291S and S447X polymorphisms: their influence on premature coronary heart disease and plasma lipids. AB - Lipoprotein lipase (LPL) plays a pivotal role in lipoprotein metabolism. Three recently described exonic polymorphisms of the gene, D9N, N291S and S447X, have been variably found to influence plasma lipids while effects on coronary heart disease (CHD) are less well documented. Two predominantly Caucasian groups were studied: CHD patients <50 years of age, with angiographically documented CHD; and a randomly recruited community control group without a history of heart disease. The 9N allele of the D9N polymorphism was present in 25 of 428 (5.8%) of Caucasian males with CHD and in seven of 291 (2.4%) of corresponding community subjects (odds ratio, 2.5; 95% confidence interval (CI), 1.1-5.9; P=0.03) and was also significantly over-represented in the Caucasian males with myocardial infarction (MI) (21 of 308 or 6.8%; odds ratio, 2.6; 95% CI, 1.1-5.9; P=0.01). The distributions of the other two polymorphisms were similar in the CHD and community groups. In multivariate models adjusted for age, sex, diabetes, body mass index, smoking, lipid levels and race, the D9N polymorphism remained significantly related to both CHD and MI, with an odds ratio >2. There were, generally, trends to more adverse fasting plasma high-density lipoprotein (HDL) cholesterol and triglycerides in carriers of the 291S and 9N alleles, and the opposite trends for triglycerides in 447X carriers. In the community group, male carriers of 291S (n=13) had significantly (20%) lower HDL cholesterol than corresponding non-carriers (n=323), 0.98+/-0.07 mmol/l (mean+/-S.E.) versus 1.22+/-0.02 mmol/l (P<0.005), while HDL cholesterol was not different in male carriers (n=8) and non-carriers (n=296) of 9N (1.23+/-0.13 mmol/l versus 1.22+/ 0.02 mmol/l). Multivariate analysis confirmed that the 291S allele carrier status conferred a significantly lower HDL cholesterol (P=0.001) and the 447X allele lower triglyceride (P<0.01) in the community group. In conclusion, LPL 9N carrier status was unequivocally related to premature CHD and to MI in males, strongly supporting recent results in older aged males. The somewhat different effects of the D9N and N291S polymorphisms on plasma lipids, and the absence of a clear effect of the N291S on CHD, raise the possibility that the effect of 9N carrier status might be mediated through effects on LPL function in addition to those influencing fasting plasma lipids. PMID- 11427212 TI - Effect of dietary omega-3 fatty acids on high-density lipoprotein apolipoprotein AI kinetics in type II diabetes mellitus. AB - The effect of a dietary fish oil supplementation on metabolism of HDL was studied in type II diabetes mellitus. Endogenous labeling of HDL-apo AI was performed using a 14 h primed infusion of D3-leucine in five diabetic patients before and 2 months after treatment with maxEPA(R). Isotopic enrichment curves were analyzed using a monoexponential function. After treatment, plasma cholesterol level remained unchanged (205.4+/-41.9 vs. 206.8+/-30.7 mg/dl, NS), whereas plasma triglycerides were decreased (155.4+/-67.9 vs. 202.6+/-32.2 mg/dl, P=0.06). Plasma apo AI was similar under maxEPA(R) (116.0+/-25.6 vs. 111.8+/-25.4 mg/dl, NS), and HDL-cholesterol and HDL-triglycerides were also not markedly changed (30.2+/-10.0 vs. 27.1+/-10 mg/dl, and 15.3+/-9.8 vs. 19.2+/-10.4 mg/dl, NS). HDL apo AI fractional catabolic rate (FCR) and absolute production rate (APR) were significantly decreased after treatment with maxEPA(R) (0.27+/-0.09 vs. 0.37+/ 0.08 pool day, P<0.05, and 12.1+/-2.8 vs. 16.1+/-3.3 mg/kg per day, P<0.05). These findings showed an effect of maxEPA(R) on kinetics of apolipoprotein AI in type II diabetes mellitus, probably linked to changes in plasma triglyceride level. PMID- 11427213 TI - Efficacy and short-term safety of a new ACAT inhibitor, avasimibe, on lipids, lipoproteins, and apolipoproteins, in patients with combined hyperlipidemia. AB - Although acyl-CoA:cholesterol acyltransferase (ACAT) inhibitors have been shown to reduce lipid levels in several animal models, the safety and lipid modifying activity of any single agent in this class has not been demonstrated in humans. The safety and efficacy of avasimibe (CI-1011), a new, unique, wholly synthetic ACAT inhibitor, was evaluated in the treatment of 130 men and women with combined hyperlipidemia and hypoalphalipoproteinemia (low levels of high-density lipoprotein cholesterol [HDL-C]). Following an 8-week placebo and dietary controlled baseline period, patients were randomly assigned to double-blind treatment with placebo, 50, 125, 250, or 500 mg avasimibe administered as capsules once daily for 8 weeks. At all evaluated doses, avasimibe treatment resulted in prompt and significant reductions (P<0.05) in plasma levels of total triglycerides (TG) and very low-density lipoprotein cholesterol (VLDL-C) with mean reductions of up to 23% and 30% respectively, apparently independent of dose. No statistically significant changes in total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), HDL-C or apolipoprotein (apo) B were detected. ApoAI levels were also unchanged on all doses of avasimibe apart from the 500 mg dosage, which was associated with a significant decrease in plasma apoAI. The relevance of this latter finding in only one dosage group is not known. All doses of avasimibe were well tolerated with no resulting significant abnormalities of biochemical, hematological, or clinical parameters. PMID- 11427214 TI - Isolation of remnant particles by immunoseparation: a new approach for investigation of postprandial lipoprotein metabolism in normolipidemic subjects. AB - Abnormal postprandial lipoproteins are associated with an increased risk for cardiovascular disease. Postprandial remnant lipoproteins were usually analyzed indirectly using retinyl esters (RE) as a chylomicron core label during an oral fat loading test. Apo B-100 containing VLDL remnants in addition to apo B-48 containing chylomicron remnants can also be directly quantified using the RLP Cholesterol Immunoseparation Assay. This recently available method uses monoclonal antibodies to apo A-I and apo B-100 to remove non-remnant lipoproteins and quantifies cholesterol in the remaining apo E-rich remnant fraction. In the present study we compared the analysis of retinyl ester with the immuno-based RLP Cholesterol (RLP-C) analysis in measuring postprandial remnant lipoproteins in healthy normolipidemic subjects. Sixteen healthy normolipidemic subjects were selected for this study. Postprandial plasma retinyl esters peaked at 5.0+/-1.2 h, whereas plasma RLP-C showed a peak significantly earlier (P<0.001) at 3.5+/ 0.6 h. In comparison, postprandial plasma TG and FFA peaked at 3.3+/-1.1 h (P<0.005 compared to retinyl esters). In conclusion, levels of RLP-C changed, during the postprandial phase, in parallel with plasma TG and FFA concentrations and peaked significantly earlier than retinyl esters. Postprandial measurements of RLP-C can be considered as a fast alternative method for the more laborious retinyl-ester analysis in clinical studies. PMID- 11427215 TI - Preactivated monocytes from hypertensive patients as a factor for atherosclerosis? AB - Recently, we reported our findings regarding the elevated secretion patterns of proinflammatory cytokines obtained from peripheral blood monocytes of hypertensive patients. To investigate the direct impact of these preactivated monocytes, the adhesion of monocytes from normal controls and hypertensive patients to vascular endothelial cell monolayers was determined spontaneously and after in vitro stimulation with either lipopolysaccharide (LPS) or angiotensin II (Ang II), with or without preincubation with the AT1 receptor antagonist eprosartan. Peripheral blood monocytes from 20 patients and 20 healthy individuals were isolated by density gradient centrifugation and plastic adherence; endothelial cells were obtained from human umbilical cords by collagenase digestion. The adhesion was determined by an assay with 51Cr radiolabeled monocytes. Oxygen species release induced by phorbol myristate acetate (PMA) as a further activation marker was analyzed for monocytes and HUVEC by chemiluminescence (CL). Spontaneous adhesion of monocytes from patients and the adhesion after stimulation with Ang II were significantly increased compared with normal controls (P<0.05). Preincubation with eprosartan diminished the adhesion in both groups to comparable levels. In monocytes, peak levels of PMA and Ang II induced CL analysis were significantly higher in patients (P<0.005). These data indicate that preactivated monocytes from hypertensives may be of pathogenic importance in atherosclerosis. PMID- 11427216 TI - Haptoglobin polymorphism as a risk factor for coronary heart disease mortality. AB - OBJECTIVES: the aim of our study was to evaluate the independent role of the haptoglobin (Hp) polymorphism as a risk factor for coronary heart disease (CHD) mortality. METHODS: within the framework of the longitudinal part of the Belgian Interuniversity Research on Nutrition and Health (BIRNH) survey, a nested case control study design was performed through matching the 107 deaths from CHD, occurring within a 10-year follow-up period, with three controls for age and gender. RESULTS: the distribution of the Hp types was found to be in Hardy Weinberg equilibrium. Conditional logistic regression analysis for matched sets revealed that the Hp polymorphism was significantly associated with CHD death. Rather surprisingly, the finding was that Hp 1-1 individuals were at doubled risk for CHD mortality compared with the others, the odds ratio being 2.09 (95% CI: 1.22-3.60). The association was independent from other classical cardiovascular risk factors and the Hp concentration, and of comparable magnitude between men and women. Moreover, evaluating the interaction term in a multiplicative model showed that the Hp type did not play a synergistic role in the prognostic value of established cardiovascular risk factors. CONCLUSION: in contrast to the findings from cross-sectionally based studies, the results from this longitudinal study show that Hp 1-1 individuals are at elevated risk for CHD mortality. PMID- 11427217 TI - Alpha-tocopherol (vitamin E) and beta-carotene supplementation does not affect the risk for large abdominal aortic aneurysm in a controlled trial. AB - Antioxidants may retard atherogenesis and limit inflammatory processes involved in aneurysm formation. We evaluated effects of alpha-tocopherol and beta-carotene supplementation on incidence of large abdominal aortic aneurysm (AAA) in a randomised, double-blind, placebo-controlled trial. Subjects (n=29133) were 50-69 years-old male smokers, participants in the Finnish alpha-Tocopherol, beta Carotene Cancer Prevention (ATBC) Study. They were randomised to receive either 50 mg/day of alpha-tocopherol, or 20 mg/day of beta-carotene, or both, or placebo in a 2x2 design. Incidence of AAA was evaluated from mortality and hospital registers. During 5.8 years of follow-up, 181 men were diagnosed with either ruptured AAA (n=77) or nonruptured large AAA treated with aneurysmectomy (n=104). Relative risk (RR) for AAA was 0.83 (95% confidence interval [CI] 0.62-1.11) among men receiving alpha-tocopherol compared with those who did not, and 0.93 (95% CI 0.69-1.24) among men receiving beta-carotene compared with those who did not. A modest though nonsignificant decrease in risk for nonruptured AAA was observed among alpha-tocopherol supplemented men (RR 0.71, 95% CI 0.48-1.04) compared with men not receiving alpha-tocopherol. For beta-carotene, RR for nonruptured AAA was 0.86 (95% CI 0.59-1.27) compared with men not receiving beta carotene. Neither antioxidant affected risk for ruptured AAA. In conclusion, long term supplementation with alpha-tocopherol or beta-carotene had no preventive effect on large AAA among male smokers. PMID- 11427218 TI - Plasma homocysteine, methylenetetrahydrofolate reductase mutation and carotid damage in elderly healthy women. AB - Plasma homocysteine (Hcy) is an independent vascular risk factor. Its remethylation to methionine is regulated by the activity of the enzyme 5,10 methylene tetrahydrofolate reductase (MTHFR). A C-to-T substitution at nucleotide 677 of the MTHFR gene is frequently associated to hyperhomocysteinemia. In this study, we evaluated the relationship among MTHFR C677T polymorphism, Hcy and some ultrasonographic parameters at the level of carotid arteries in 120 elderly women with normal ECG, normal blood pressure values, total cholesterol <250 mg/dl, normal glucose tolerance, normal albumin excretion rate. In all subjects, we measured Hcy by HPLC, MTHFR mutation by polymerase chain reaction followed by HinfI digestion and intima-media thickness (IMT), peak velocity of the systolic flow (SP(V)), end-diastolic velocity (ED(V)) and resistance and pulsatility indexes of intracranial circulation (RI and PI) by ultrasound imaging. Twenty eight women were homozygotes for the wild type allele (Ala/Ala), 72 were heterozygotes (Ala/Val) and 20 were homozygotes for the mutation (Val/Val). Groups were comparable for age, blood pressure values and plasma lipid levels. Hcy was higher in Val/Val group; moreover, after adjustment for confounding factors, Val/Val had significantly greater IMT and ED(V) (P<0.001 and P<0.05, respectively). Logistic analysis revealed that Val/Val genotype was the strongest risk factor for IMT (OR 30.8, 95% CI 2.82-335.6). Our results show that, in elderly healthy women, Val/Val homozygosity for C677T mutation in MTHFR gene could identify subjects at risk for asymptomatic carotid atherosclerotic impairment. PMID- 11427219 TI - Remnant-like lipoprotein particle cholesterol concentration and progression of coronary and vein-graft atherosclerosis in response to gemfibrozil treatment. AB - Remnant lipoproteins such as chylomicron and very low density lipoprotein (VLDL) remnants have been implicated in the progression of coronary atherosclerosis. Recently, a novel method for the determination of the remnant-like lipoprotein particle cholesterol (RLP-C) concentration was developed based on immunoaffinity separation of plasma. The compositional characteristics of RLP are strikingly similar to those of postprandially modified VLDL. In addition, the method also detects chylomicron remnants. We investigated the relationship between the plasma RLP-C concentration and the angiographic outcome of the 2-year, randomised, placebo-controlled Lipid Coronary Angiography Trial (LOCAT), which used gemfibrozil as lipid lowering agent. The RLP-C response to gemfibrozil treatment has not been described before. Gemfibrozil reduced the median RLP-C concentration by 34%. The on-treatment RLP-C concentration was significantly associated with the progression of the minimum lumen diameter (MLD) (P<0.004). The plasma levels of RLP-C as well as the change in response to treatment was closely associated with plasma triglycerides and the association between on-treatment RLP-C concentration and progression of MLD was not independent of plasma triglycerides. A significant relation was seen between RLP-C and the occurrence of new lesions in vein grafts. Subjects with one new lesion had an approximately 25% higher on treatment RLP-C concentration and the four patients showing two new lesions had a 100% higher RLP-C concentration than patients without vein graft stenosis. A total of 19 out of 23 subjects having one new lesion, and all four patients showing two new lesions, were assigned to the placebo group. We conclude that the RLP-C concentration, which is likely to reflect the plasma cholesterol contained in postprandially modified VLDL and chylomicron remnants, is strongly associated with angiographically verified progression of focal coronary atherosclerosis, and that lowering of RLPs prevents vein graft stenosis. PMID- 11427220 TI - Fibrinogen and C-reactive protein on admission as markers of final infarct size after primary angioplasty for acute myocardial infarction. AB - BACKGROUND: In acute myocardial infarction (AMI) treated conservatively or with thrombolysis, marked increases of C-reactive protein (CRP) and fibrinogen have been observed. No data are however available concerning a possible relation between CRP and fibrinogen levels on admission and markers of infarct size after obtaining thrombolysis in myocardial infarction (TIMI) flow III by primary angioplasty. METHODS: We studied 34 patients with a first AMI (29 men, mean age 54+/-11 years) who were treated with primary angioplasty (TIMI flow III in all patients, no concomitant treatment with glycoprotein IIb-IIIa antagonists) within 6 h of onset of pain. CRP and fibrinogen levels on admission were determined and related to the following markers of infarct size: peak creatine kinase MB (CKMB) levels, radionuclide left ventricular ejection fraction (LVEF) at discharge and thallium-201 single-photon emission computed tomography (SPECT) infarct size at 1 month. RESULTS: Median CRP levels were 0.4 mg/dl (range 0.09-3 mg/dl), median fibrinogen levels 412 mg/dl (range 198-679 mg/dl), mean CKMB was 178+/-151 U/l, mean LVEF 52+/-8% and mean thallium-201 infarct size 7+/-6%. Although CRP levels were related to fibrinogen levels on admission (r=0.56, P=0.002), only fibrinogen levels were related to markers of infarct size (r=0.58, P=0.001 for CKMB, r= 0.44, P=0.01 for LVEF and r=0.64, P=0.001 for thallium-201 infarct size). No relation was found between CRP or fibrinogen levels on admission and the extent of coronary artery disease or the myocardial area at risk. In multiple regression analysis, the relation between fibrinogen and markers of infarct size was independent of CRP levels and the duration of pain on admission. CONCLUSIONS: These findings indicate a relation between fibrinogen levels on admission and myocardial infarct size in patients treated with primary angioplasty for AMI. This relation seems to be independent of CRP levels and the duration of pain on admission. If confirmed in larger patient populations, fibrinogen levels on admission could have an important value for risk stratification and more aggressive reduction of infarct size in patients who are treated with primary angioplasty. PMID- 11427221 TI - Independent relationship between microalbuminuria and plasminogen activator inhibitor-1 activity (PAI-1) activity in clinically healthy 58-year-old men. AB - The aim of this cross-sectional study was to investigate the relationship between low-grade albuminuria (microalbuminuria) and factors of the coagulation- and fibrinolysis systems in 104 clinically healthy 58-year-old men recruited from the general population. Urinary albumin excretion was significantly associated with body mass index, systolic and diastolic blood pressure, plasminogen activator inhibitor (PAI)-1 activity, tissue plasminogen activator (tPA) antigen, tPA activity (negatively) and protein S (P<0.05). There were no associations between urinary albumin excretion and antithrombin III, fibrinogen, protein C, thrombin/antithrombin factor or von Willebrand factor. In multiple regression analysis urinary albumin excretion was independently and significantly associated with PAI-1 activity and systolic blood pressure (P<0.05). In conclusion we report that urinary albumin excretion was independently and significantly associated with PAI-1 activity in clinically healthy 58-year-old men. This relationship may contribute to the previously reported increased cardiovascular morbidity in subjects with microalbuminuria. PMID- 11427222 TI - Pericardial fat accumulation in men as a risk factor for coronary artery disease. AB - An increment of abdominal visceral fat accumulation has been reported to be a coronary risk factor. We determined the predictive power of pericardial fat (Pfat) accumulation as intra-thoracic visceral fat, in the diagnosis of coronary artery disease (CAD). Among 251 (181 non-obese [body mass index<25], 70 obese [body mass index> or =25]) Japanese male patients who underwent computed tomography (CT), 128 (90 non-obese, 38 obese) patients were suffering from CAD. Pfat volume was determined by the sum of cross-sectional images 1cm thick from the atrial appendage to the apex over the diaphragm. Abdominal visceral fat (Vfat) and subcutaneous fat (Sfat) areas were measured by a single scan at the L4 L5 region. Pfat was most associated with Vfat in body fat distribution. In non obese patients, Pfat was most associated with CAD among the various risk factors including body fat distribution. Moreover, Pfat was the strongest independent variable for the severity of CAD, determined by coronary angiogram. This result showed that pericardial fat accumulation was a stronger coronary risk factor than the other body fat distributions in non-obese men. PMID- 11427223 TI - The atherogenic lipoprotein phenotype: small dense LDL and lipoprotein remnants in nephrotic range proteinuria. AB - The dyslipidaemia in nephrotic-range proteinuria is believed to contribute to the increased atherogenesis associated with the condition. Excess small dense low density lipoprotein (LDLIII) contributes to this risk. Lipoprotein remnants (RLP) may also be implicated but have not been studied in this population. We measured the plasma concentration of low density lipoprotein (LDL) subfractions (by density gradient ultracentrifugation), RLP (by immunoaffinity gel), very low density lipoprotein (VLDL) subfractions, post heparin lipases and cholesteryl ester transfer protein (CETP) activity in 27 patients with glomerular disease and albuminuria >2.0g. These were compared with 27 age and sex matched controls. Proteinuric patients had increased LDLIII concentration (patients 182 (84:267) vs. controls 31 (27:62); P<0.0001) with reduced lighter LDLI (36 (24:43) vs 69 (46:101); P<0.0005) and LDLII (124 (79:220) vs 178 (129:236); P<0.04, all mg/dl, median+interquartile range). RLP-cholesterol (RLP-C) and triglyceride (RLP-TG) were increased in proteinuric patients (RLP-C 18.9 (11.0:26.9) vs 7.7 (6.0:8.8); P<0.0001, RLP-TG 35.8 (11.8:54.7) vs. 7.2 (4.3:10.0); P<0.0001, all mg/dl). Increased LDLIII and RLP were independent of renal function. VLDL(1) and VLDL(2) concentrations were increased by 258 and 260% (both P<0.0001). CETP activity was increased by 46% (P<0.005). Lipoprotein and hepatic lipase activities did not differ from control values. LDLIII concentration (r(2)=45.7%, P<0.001), RLP-C (r(2)=85.2%, P<0.001) and RLP-TG (r(2)=87.5%, P<0.001) all correlated positively with plasma triglyceride. Moreover, increased LDLIII was associated with both RLP C (r(2)=31.3%, P<0.002) and RLP-TG (r(2)=33.6%, P<0.002). Excess LDLIII and RLP are present in nephrotic-range proteinuria and add to the spectrum of cardiovascular risk factors present in proteinuric patients. Increases in LDLIII and RLP are closely related to plasma triglyceride. The association between excess RLP and LDLIII suggests that RLP contribute to the increased atherogenicity attributed to the atherogenic lipoprotein phenotype. PMID- 11427224 TI - Glucocorticoid receptor Bcl I variant is associated with an increased atherogenic profile in response to long-term overfeeding. AB - The effect of the glucocorticoid receptor (GRL) gene Bcl I polymorphism on body composition and metabolic changes in response to overfeeding was studied. Twenty four men (mean age 21+/-2 years) who constituted 12 pairs of identical twins ate a 4.2 MJ/day energy surplus, 6 days a week, during a period of 100 days. The GRL Bcl I marker was identified by Southern Blot technique. Total body fat was assessed by hydrodensitometry and abdominal fat areas were measured by computed tomography. Fasting plasma glucose and insulin during an oral glucose tolerance test (OGTT) were assayed. The insulin and glucose areas were computed using the trapezoidal method. Triglyceride and cholesterol concentrations in plasma and lipoprotein fractions were determined enzymatically. The results showed that overfeeding induced a greater increase in body weight (p=0.002) in the 2.3/2.3 kb (n=12) than in the 4.5/2.3 kb (n=12) subjects. In addition, plasma levels of total (p=0.007) and low-density lipoprotein (LDL) cholesterol (p=0.003), as well as systolic blood pressure (p=0.036) increased more in the 2.3/2.3 kb than in the 4.5/2.3 kb subjects. The 2.3/2.3 kb genotype was also associated with a greater increase in abdominal visceral fat (p=0.040) compared to the 4.5/2.3 kb genotype. In conclusion, 2.3/2.3 kb subjects of the GRL Bcl I polymorphism experience greater increases in body weight, blood pressure and cholesterol levels as well as visceral fat than 4.5/2.3 kb subjects in response to overfeeding. These data suggest that overfeeding induces an atherogenic profile in subjects who are homozygotes for the 2.3 kb allele. PMID- 11427225 TI - Leukocyte count, diabetes mellitus and age are strong predictors of stroke in a rural population in southern Italy: an 8-year follow-up. AB - Stroke incidence rates in the Mediterranean area are higher compared to northern European countries. In this study, we present the 8-year prospective data from a small rural Sicilian town. This population, consisting of 1351 subjects (622 males and 729 females), is homogeneous for ethnic background with traditional healthy dietary habits and shows low cholesterol mean levels. We found that the risk of stroke was significantly associated with the record of at least one previous neurological symptom (PNS), such as lack of strength, loss of vision or speech or possible drop attacks, and high hematocrit in males, and to high body mass index (BMI) and waist-hip ratio (WHR), diabetes, hypertension, high leukocyte count in females. We also documented age-related differences: stroke was associated in younger subjects (age<65 years) with diabetes, high BMI, high uric acid levels and in older patients (age>/=65 years) with high WHR, hypertension, diabetes, PNS, leukocyte count and hematocrit above the 95th percentile. Multivariate analysis demonstrated an independent association between stroke and age, diabetes, leukocyte count, hypertension and PNS. In conclusion, in this rural Sicilian population, the incidence rate of stroke is 1.72 cases per 1000/year in the subjects between 40 and 75 years of age. The risk factors associated with stroke are different in younger and older subjects. Leukocyte count, as an expression of an undergoing inflammatory process, may have a relevant role at least in the elderly. PMID- 11427226 TI - Effects of gender, hepatic lipase gene polymorphism and type 2 diabetes mellitus on hepatic lipase activity in Chinese. AB - Genetic variation in the hepatic lipase (HL) gene (LIPC) promoter is an important determinant of HL activity in Caucasians. As HL activity is increased in patients with type 2 diabetes mellitus, we have investigated whether the -514 C-to-T polymorphism acted independently of type 2 diabetes to regulate HL activity. The frequency of this polymorphism and its effect on plasma HL activity and lipids were examined in 203 Chinese patients with type 2 diabetes and 205 controls. The frequency of the T allele was 0.343 and 0.376 in male and female diabetic patients, respectively, compared with 0.371 and 0.372 in male and female controls. The effect of LIPC genotype on HL activity was similar between men and women, and between diabetic patients and non-diabetic controls, with the lowest HL activity being found in those subjects with the TT genotype. On multivariate analysis, gender, LIPC genotype, the presence of type 2 diabetes and body mass index were independent predictors of HL activity, accounting for 22, 9, 5 and 3%, respectively, of the variance in HL activity (whole model adjusted R(2)=0.39, P<0.0001). The T allele was associated with higher high-density lipoprotein in the controls but not in the diabetic patients, and no associations were found between LIPC genotype and low-density lipoprotein subfractions in either groups. In conclusion, despite the higher frequency of the T allele in Chinese than in Caucasians, gender was the best predictor for HL activity, with LIPC gene polymorphism and type 2 diabetes making relatively smaller contributions to the variation in HL activity. PMID- 11427227 TI - Lipoprotein (a) and development of intermittent claudication and major cardiovascular events in men and women: the Edinburgh Artery Study. AB - Lipoprotein (a) may be an important risk factor for atherosclerosis. It is widely accepted that lipoprotein (a) levels are raised in patients with coronary heart disease, but there is some doubt about the causality of the relationship. In addition, little is known about the relationship between lipoprotein (a) and either stroke or peripheral arterial disease, nor about the role of lipoprotein (a) in women. Subjects aged 55-74 years (n=1592) were selected at random from 11 general practices in Edinburgh, Scotland and followed up for 5 years. The incidences of myocardial infarction, intermittent claudication and stroke were 13.4, 9.4 and 3.7%, respectively. Raised lipoprotein (a) levels at baseline were associated with an increased risk (95% confidence interval) of myocardial infarction RR 1.15 (1.00, 1.32), intermittent claudication RR 1.32 (1.10, 1.57) but not significantly for stroke RR 1.24 (0.93, 1.64). This increased risk persisted for intermittent claudication after adjustment for baseline cardiovascular disease and other risk factors RR 1.20 (1.00, 1.43), but for myocardial infarction became non-significant RR 1.06 (0.91, 1.23). The risk of disease associated with raised lipoprotein (a) was slightly higher in women than in men, especially for intermittent claudication (men RR 1.09 (0.87, 1.36) compared to women RR 1.37 (1.01, 1.87)). In conclusion, we found that lipoprotein (a) was an independent predictor of cardiovascular events in both sexes. The association between lipoprotein (a) and cardiovascular events may have been stronger in women than in men, and for peripheral arterial disease than myocardial infarction and stroke. PMID- 11427228 TI - Activation of metalloproteinase-2, loss of matrix scleroprotein content and coronary artery calcification. AB - Plaques from the coronary arteries of explanted hearts showed massive calcification (15-fold increase) with a loss of scleroproteins (-36%), an increase in the collagen to elastin ratio (twofold) and activation (+15%) of matrix metalloproteinase-2 (MMP-2). Plaque-free portions of the coronary artery gave results similar to those obtained with the internal mammary artery. There was a significant correlation between plaque calcification and MMP-2 activation, suggesting that the two processes may be linked. PMID- 11427229 TI - Part III. Direct enzymatic esterification of lactic acid with fatty acids. AB - Lipase catalyzed esterification reactions between lactic acid and several fatty acids have been studied. Difficulties arise in esterifying lactic acid because of the potential for this substance to act both as an acyl donor and as a nucleophile. These difficulties were minimized via strategies which greatly increased the yield of the desired ester. Use of the companion fatty acid in excess with respect to lactic acid in an apolar solvent (n-hexane) in which the lactic is not completely dissolved has been employed to minimize the potential for lactic acid to act as an acyl donor in a self-polymerization reaction.Beneficial and sinergistic effects of both silica gel and molecular sieves on conversion to the desired product are described. However, careful control of the amount of molecular sieves used is required. This fact is a consequence of two opposing effects of this material: i.e. adsorption of both lactic acid and water from the reaction mixture. For reaction between caprylic and lactic acids, use of an excessive amount of enzyme reduces the extent of conversion to 2-O-caproyl-lactic acid.A very pure ester of the L-enantiomer (optical rotation of [alpha]D(25) = -23.5) can be prepared in n-hexane using a four fold excess of caprylic acid and Candida antarctica lipase. Optimum reaction conditions lead to 35% yield of 2-O-caproyl-lactic acid, a result which is close to the maximum yield that can be enantioselectively obtained from commercial grade lactic acid (68 mole per cent monomer). PMID- 11427230 TI - Effects of growth medium composition, iron sources and atmospheric oxygen concentrations on production of luciferase-bacterial magnetic particle complex by a recombinant Magnetospirillum magneticum AMB-1. AB - Growth conditions for mass production of luciferase-bacterial magnetic particles (BMPs) by a recombinant Magnetospirillum magneticum AMB-1 were investigated in a pH-regulated fed-batch culture system. Enrichment of growth medium with L cysteine, yeast extract and polypeptone enhanced both bacterial growth and BMP production. The presence of L-cysteine in the medium was useful for induction of cell growth. Strict anaerobic conditions led to a prolonged lag phase and limited the final cell density. Trace oxygen enhanced cell growth with increasing BMP production. As iron sources, ferrous sulfate and ferric gallate dramatically enhanced BMP yield as compared with ferric quinate, an iron chelate conventionally used. The optimized conditions increased cell density to 0.59 +/- 0.03 g cell dry weight/liter and BMP production to 14.8 +/- 0.5 mg dry weight/liter in fed-batch culture for four days. PMID- 11427231 TI - Regioselective biotransformation of the dinitrile compounds 2-, 3- and 4 (cyanomethyl) benzonitrile by the soil bacterium Rhodococcus rhodochrous LL100 21. AB - The cyanomethyl benzonitrile compounds used for this study contain two cyano groups: a -CH(2)CN side chain, plus a cyano group attached to the benzene ring. The ortho, meta and para -CH(2)CN substituted compounds were biotransformed using whole cell suspensions of the bacterium Rhodococcus rhodochrous LL100-21. The bacterium had previously been grown on the mono-nitrile compounds propionitrile, benzonitrile or acetonitrile, inducing the formation of nitrile hydrolyzing enzymes.Suspensions of R. rhodochrous LL100-21 that had been grown on propionitrile or benzonitrile converted the aliphatic group of 2-(cyanomethyl) benzonitrile (a) to the corresponding carboxylic acid, 2-(cyanophenyl) acetic acid (d) with excellent recovery of the product and no evidence for any other products. Conversely, when grown on acetonitrile the bacterium converted 2 (cyanomethyl) benzonitrile (a) to the amide derivatives 2-(cyanophenyl) acetamide (k) and 2-(cyanomethyl) benzamide (l) but only in low yields.Biotransformations of 3-(cyanomethyl) benzonitrile (b) and 4-(cyanomethyl) benzonitrile (c), by suspensions of bacteria that had been grown on benzonitrile or propionitrile, resulted in hydrolysis of the aromatic nitrile to produce 3- and 4-(cyanomethyl) benzoic acid (j) and (m), respectively, both with a high yield. Low concentrations of other products were also detected, for example the diacids 3- and 4-(carboxyphenyl) acetic acid (h) and (i).When the bacterium was grown on acetonitrile it could biotransform 3- and 4-(cyanomethyl) benzonitrile (b) and (c) to different products indicating less regiospecificity by the nitrile hydratase enzyme.Comparison of the initial rates of conversion of the aliphatic cyano side chain of 2-(cyanomethyl) benzonitrile (a) and other substituted benzonitriles indicated that electronic effects did not affect the initial rate of the reaction as they would require transmission through an SP(3) methylene carbon atom. PMID- 11427232 TI - Effect of contact time and inhibitor concentration on the affinity mediated adsorption of cells to surfaces. AB - Cell detachment by shear stress under conditions of laminar flow was used to investigate the effect of incubation time and soluble binding competitors on affinity mediated cell/surface interactions. Fractional attachment between yeast and a Concanavalin A (Con A) coated surface was studied as a function of adhesion time prior to exposure to shear in a parallel plate flow chamber. Two, four and sixteen hours adhesion times gave rise to significantly different fractional attachment profiles, with four hours giving greater cell retention.The effect of dextran as a competitive displacer of pre-attached cells was also examined using a number of exposure regimes. While the presence of dextran in the displacement buffer led to higher fractional displacement of pre-attached cells, this effect was magnified if an equilibration period between dextran solution and pre attached cells was allowed before detachment was attempted. The decline in fractional attachment increased with incubation time up to 30 min, with longer periods resulting in a smaller effect. Pre-incubation of the Con A surface with dextran prior to the introduction of cells led to a 60% reduction in attachment.Attempts to determine critical shear values were complicated by the presence of a tightly bound cell fraction of approximately 15% that was not removed at the highest shear values used. PMID- 11427233 TI - Inactivation of lignin peroxidase during oxidation of the highly reactive substrate ferulic acid. AB - Ferulic acid (4-hydroxy-3-methoxycinnamic acid) (FA) was found to be a highly reactive substrate for lignin peroxidase (LIP), exhibiting a k(cat) of 41.7 s( 1). Despite the high reactivity, two modes of inactivation prevailed during the oxidation of FA. The first, H(2)O(2)-dependent inactivation, was evidenced by incomplete substrate oxidation and accumulation of LIP compound III (LIPIII), even at relatively low H(2)O(2) concentrations. This was attributed to the high turnover rate along with the inability of FA to revert LIPIII to the native state, as evidenced by pre-steady-state kinetics. H(2)O(2)-dependent inactivation could be avoided by inclusion of veratryl alcohol (VA), which efficiently reverts LIPIII to the native state. However, VA also mediated FA oxidation, and significantly decreased the reaction rate, which is unlike for previously reported VA-mediated reactions. The second mechanism of LIP inactivation was attributed to binding of phenoxy radicals or oxidation products to the enzyme and its extent directly correlated with the amount of FA consumed. This inactivation could be considerably suppressed by inclusion of gelatin. Therefore, during the oxidation of highly reactive phenolics, different kinds of protectors are required for efficient oxidation and maintaining LIP activity over time. This is of importance when considering emerging biotechnological applications for LIP. PMID- 11427234 TI - Hexose oxidase from Chondrus crispus: improved purification using perfusion chromatography(*). AB - 1An improved method for purifying hexose oxidase (D-hexose: O(2) 1 oxidoreductase, EC 1.1.3.5) from the marine red alga Chondrus crispus is described for obtaining enzyme suitable for structural characterization and use in bioconversion of lactose to lactobionic acid. This involved extracting enzyme from finely ground lyophilized tissue in sodium phosphate buffer (pH 7) containing 20% ammonium sulfate, eliminating the previously used solvent extraction and protease treatments, and by applying Poros perfusion chromatography media to achieve rapid separations of high resolution. Primary separation of contaminating phycobiliproteins and carrageenans was achieved using Poros DEAE-50. Sequential HPLC purification steps using Poros HP2 and Poros HQ were followed by Sephacryl S200 h chromatography. Enzyme activity was determined with a peroxidase-coupled assay using 2,2'-azino-bis (3-ethylbenzthiazoline-6 sulfonic acid) substrate. A final specific activity of 69 U/mg was obtained, representing a 100-fold purification with an activity recovery of about 10%. A native size of approximately 117,000 Da was determined by size exclusion chromatography, and SDS-PAGE revealed the presence of 38,000 and 29,000 Da polypeptides that appear to be derived from a 65,000 Da subunit. Further properties of the enzyme are described. PMID- 11427235 TI - Parameters that determine virus adsorption kinetics: toward the design of better infection strategies for the insect cell - baculovirus expression system. AB - High productivities of bioprocesses involving viruses can be attained through infection strategies based on adequate understanding of parameters ruling cell virus interactions. Two factors that affect virus binding and infection efficiency were studied: the utilization of an adsorption step, where infection volume at constant cell/virus ratio was varied; and the concentration of fetal bovine serum (FBS). The insect cell-baculovirus expression system and recombinant protein VP4 of rotavirus were used as models. Virus binding kinetics were adequately described by a sigmoidal response curve. The adsorption step, with or without FBS, increased virus attachment rate, whereas it increased bound virus at equilibrium only in FBS-free infections. A first-order dependance of virus attachment on cell concentration was found above 5 x 10(6) cell/mL in infections with 10% FBS. Addition of 10% FBS decreased maximum bound baculovirus and binding rate by as much as 3 times and VP4 concentration up to 4 times. In contrast, heat inactivation of FBS increased bound virus from 20% to over 90%, an increase of 1.4 times compared to FBS-free infections. A direct linear relation was found between attached virus and maximum VP4 concentration for the different FBS concentrations tested, indicating that baculovirus-cell attachment was the limiting step for recombinant protein production. Interestingly, virus progeny accumulation was not affected by differences in virus binding. In conclusion, infection strategies aimed at increasing productivity should be performed at high cell concentrations and without FBS, or with heat-inactivated FBS. PMID- 11427236 TI - Dechlorination of chlorophenols using extracellular peroxidases produced by streptomyces albus ATCC 3005. AB - Streptomyces albus ATCC 3005 was found to produce higher levels of extracellular peroxidase activity (3.420 U mg(-1)) than previously reported for any other actinomycete. Maximum peroxidase activity was obtained after 72 h of incubation at a temperature of 30 degrees C in a liquid medium (pH 7.6) containing (in w/v) 0.8% to 0.9% oat spelts xylan and 0.6% yeast extract, corresponding to a C:N ratio of around 8.4:1. Characterization of the peroxidases revealed that the optimal temperature for peroxidase activity, using the standard 2,4 dichlorophenol (2,4-DCP) assay was 53 degrees C, when the enzyme reaction was performed at pH 7.2. A study of the effect of temperature on the stability of peroxidase over time, showed that the enzyme was stable at 40 degrees C, with a half-life of 224 min, while at higher temperatures the stability and activity was reduced such that at 50 degrees C and 70 degrees C the half-life of the enzyme was 50 min and 9 min respectively. The optimum pH for the activity of the enzyme occurred between pH 8.1 and 10.4. In terms of substrate specificity, the peroxidase was able to catalyze a broad range of substrates including 2,4-DCP, L 3,4-dihydroxyphenylalanine (L-DOPA), 2,4,5-trichlorophenol and other chlorophenols in the presence of hydrogen peroxide. Ion exchange chromatography was used to confirm that the enzyme was able to release chloride ions from a range of chlorophenols. PMID- 11427237 TI - Purification and properties of a high-molecular-weight, alkaline exopolygalacturonase from a strain of Bacillus. AB - 1An exopolygalacturonase [exo-PG; poly (1,4-alpha-D-galacturonide) digalacturonohydrolase, EC 3.2.1.82] was found in a culture of Bacillus sp. strain KSM-P576. The purified exo-PG had a molecular weight of approximately 115,000 and an isoelectric point of pH 4.6. The N-terminal amino acid sequence was Thr-Glu-Val-Ser-Pro-Lys-Ser-Pro-Ala-Ser-Pro-Val. Maximum activity toward polygalacturonic acid (PGA) was observed at 55 degrees C and pH 8.0 in 100 mM Tris-HCl buffer. The exo-PG was quite stable in various pH buffers between pH 6 and 12 when incubated at 30 degrees C for 1 h. Mg(2+,) Mn(2+,) Pd(2+) and Ca(2+) ions stimulated the enzyme activity. The exo-PG released digalacturonic acid from PGA, tri-, tetra-, and penta-galacturonic acids. The apparent K(m) values for oligogalacturonic acids were almost identical, and k(cat) values increased with the chain length of the substrates. PMID- 11427238 TI - Evidence for a non-phosphorylated route of galactose breakdown in cell-free extracts of Aspergillus niger. AB - Aspergillus niger could utilize D-galactose as sole source of carbon. Cell-free extracts of D-galactose-grown mycelia were able to catalyze the oxidation of D galactose to D-galactonic acid-gamma-lactone (GalA-gamma-lact) in the presence of NAD, followed by the appearance of 2-keto-3-deoxy-D-galactonate (KDGal), pyruvate and glyceraldehyde. From 10 &mgr;moles only 6.6 &mgr;moles of GalA-gamma-lact were disappeared after 60 min of reaction indicating the presence of GalA-gamma lactonase. Identification of GalA-gamma-lact was achieved by ascending paper chromatography. KDGal, pyruvate and glyceraldehyde were also chromatographically identified in the reaction mixture containing D-galactonate which suggests that D galactonate is degraded into pyruvate and glyceraldehyde via the intermediate formation of KDGal. Such reactions are supposed to be catalyzed by an inducible D galactonate dehydratase and a constitutive KDGal aldolase. The amount of KDGal, pyruvate and glyceraldehyde were found to be almost equivalent and the equilibrium of the reaction being toward the formation of KDGal. The apparent equilibrium constant (K(eq)) was calculated and found to be 0.5 x 10(-3) M. Results also proved the reversibility of the reaction catalyzed by KDGal aldolase of A. niger. In the light of the findings obtained from the degradation of D galactose by cell-free extracts of A. niger grown on D-galactose and D galactonate a nonphosphorolytic pathway was suggested to be operative for the degradation of D-galactose in extracts of A. niger. PMID- 11427239 TI - Use of a quartz crystal impedance technique to monitor the degradation of chondroitin sulfate by hyaluronidase. AB - A new quartz crystal impedance hyaluronidase sensing technique, which is based on the changes in viscosity and density during the enzymatic hydrolysis of chondroitin sulfate (CS) by hyaluronidase (HAse), is established in this paper. The variations of equivalent circuit parameters of the piezoelectric quartz crystal (PQC) during the enzymatic degradation are discussed. The initial hydrolysis rate of CS by HAse is obtained from the changes in viscosity and density with incubation time. Kinetic parameters (the Michaelis constant K(m) and the maximum hydrolysis rate V(max)) of the degradation process are estimated by using a linear method of Lineweaver-Burk plot. K(m) is 2.73 +/- 0.20 mg.ml(-1) and V(max) is -(5.43 +/- 0.38) x 10(-4) kg.m(-2).s(-1/2).min(-1.) PMID- 11427240 TI - Isolation and characterization of a thermostable endo-beta-glucanase active on 1,3-1,4-beta-D-glucans from the aerobic fungus talaromyces emersonii CBS 814.70. AB - A novel endoglucanase active on 1,3-1,4-beta-D-glucans was purified to apparent homogeneity from submerged cultures of the moderately thermophilic aerobic fungus Talaromyces emersonii CBS 814.70. The enzyme is a single subunit glycoprotein with M(r) and pI values of 40.7 +/- 0.3 kDa and 4.4, respectively, and an estimated carbohydrate content of 77% (w/w). The purified beta-glucanase displayed activity over broad ranges of pH and temperature, yielding respective optima values of pH 4.8 and 80 degrees C. This enzyme was markedly thermostable with 15% of the original activity remaining after incubation for 15 min at 100 degrees C. Substrate specificity studies revealed the identity of the enzyme to be a 1,3-1,4-beta-D-glucanase. Identical K(m) values (13.38 mg.ml(-1)) were obtained with lichenan and BBG, while the V(max) value with lichenan (142.9 IU.mg(-1)) was approximately twice the value obtained with BBG (79.3 IU.mg(-1)). Time-course hydrolysis of barley-beta-glucan did not proceed linearly with respect to time indicating an 'endo' or more processive action for the enzyme. HPAEC fractionation of the products of hydrolysis yielded a range of oligosaccharides, with cellobiose, cellotriose and cellotetraose being the predominant oligosaccharide products. PMID- 11427241 TI - Monitoring the expression and purification of recombinant proteins by MALDI-TOF mass spectrometry. AB - 1Samples coming from biologic sources usually contain several contaminants that interfere seriously with Mass Spectrometry (MS) measurements. In this paper we report the application of MALDI-TOF MS to monitor recombinant protein expression and purification. The technique is based on the use of a C18 resin to clean and concentrate proteins in batch. The utility of this method is demonstrated for samples coming from different bacterial cultures expressing secreted and intracellular proteins ranging from 4 to 53 kDa. MALDI-TOF MS of peptide and proteins can be accomplished directly from complex bacterial cultures or from any purification step in a few minutes using the conventional stainless steel sample targets, allowing for a nearly instantaneous monitoring of the nature and integrity of recombinant expression products. PMID- 11427242 TI - Somatic complaints among pregnant women cared for in obstetrics: normal pregnancy or depressive and anxiety symptom amplification revisited? AB - Despite the extensive research documenting the significance of medically unexplained somatic symptoms in primary care patients, few studies have examined somatic symptoms as a predictor of depressive and anxiety disorders among pregnant women cared for in Obstetrics. We utilized the Primary Care Evaluation of Mental Disorders Patient Health Questionnaire (PHQ) to assess current depressive and anxiety disorders and self-reported somatic symptoms among 186 women receiving prenatal care. We examined the bivariate relationships between depressive and anxiety disorders and mean number of somatic symptoms. Linear regression analyses assessed the unique association between maternal depression, anxiety and somatic symptoms, while controlling for selected demographics and maternal medical risk. Twenty three percent (N=43) of women met screening criteria for depressive and/or anxiety disorders. Women with depression and/or anxiety were significantly more likely to report somatic symptoms (mean=7.1, SD=2.6) compared to women without depression or anxiety (mean=5.0, SD=2.6) [t(df)=4.54(184), P<.001]. This association persisted in multivariate models. Our findings suggest that antenatal depressive and anxiety disorders are associated with an amplification of physical symptoms of pregnancy. Eliciting and tracking somatic symptoms during prenatal visits could potentially improve detection of depressive and anxiety disorders in the obstetrical sector. PMID- 11427243 TI - Collaborative interventions for physically injured trauma survivors: a pilot randomized effectiveness trial. AB - Posttraumatic behavioral and emotional disturbances occur frequently among physically injured hospitalized trauma survivors. This investigation was a pilot randomized effectiveness trial of a 4-month collaborative care intervention for injured motor vehicle crash and assault victims. As surgical inpatients, intervention subjects (N=16) were assigned to a trauma support specialist who provided counseling, consulted with surgical and primary care providers, and attempted postdischarge care coordination. Control subjects (N=18) received usual posttraumatic care. For all participants, posttraumatic stress disorder (PTSD) and depressive symptoms, episodic alcohol intoxication, and functional limitations were evaluated during the hospitalization and 1 and 4 months postinjury. Study logs and field notes revealed that over 75% of intervention activity occurred in the first month after the trauma. One-month post-trauma intervention subjects when compared to controls demonstrated statistically significant decreases in PTSD symptoms as well as a reduction in depressive symptoms. However, at the 4-month assessment, intervention subjects evidenced no significant improvements in PTSD and depressive symptoms, episodic alcohol intoxication, or functional limitations. Future larger scale trials of stepped collaborative care interventions for physically injured trauma survivors are recommended. PMID- 11427244 TI - Consultation-Liaison psychiatric service delivery: results from a European study. AB - The reported findings of the European Consultation-Liaison Workgroup (ECLW) Collaborative Study describe consultation-liaison service delivery by 56 services from 11 European countries aggregated on a C-L service level. During the period of 1 year (1991), the participants applied a standardized, reliability tested method of patient data collection, and data were collected describing pertinent characteristics of the hospital, the C-L service, and the participating consultants. The consultation rate of 1% (median; 1.4% mean) underscores the discrepancy between epidemiology and the services delivered. The core function of C-L services in general hospitals is a quick, comprehensive emergency psychiatric function. Reasons to see patients were the following. deliberate self-harm (17%), substance abuse (7.2%), current psychiatric symptoms (38.6%), and unexplained physical complaints (18.6%) (all means). A significant number of patients are old and seriously ill. Mood disorders and organic mental disorders are most predominant (17.7%). Somatoform and dissociative disorders together constitute 7.5%. C-L services in European countries are mainly emergency psychiatric services and perform an important bridge function between primary, general health, and mental health care. PMID- 11427245 TI - Physicians' low detection rates of alcohol dependence or abuse: a matter of methodological shortcomings? AB - Previous research may have underestimated physicians' detection rates of alcohol dependence or abuse because case findings have been based on screening questionnaires instead of using in-depth diagnostic criteria and detection rates have been assessed by analyzing patient records instead of directly interviewing the physician. To test this hypothesis, consecutive patients of a general hospital (N=436) and of 12 randomly selected general practices (N=929) were examined. A two-step diagnostic procedure included screening questionnaires and a diagnostic interview (SCAN). The analysis compares detection rates based on methods used in previous studies to data using more precise methods. Physicians' detection rates ranged from 37.0% to 88.9% in the general hospital and from 11.1% to 74.7% in general practices depending on methods used. The physicians' detection rates could be improved by 10% (general hospital) and 20% (general practice) through the additional use of a screening questionnaire. Of those patients assessed by the physicians as problem drinkers in the general hospital, 13.9% were referred to an addiction consultation-liaison service. Data reveal that physicians' abilities to detect problem drinkers have been underestimated. Routine screening procedures could play a major role in improving detection rates and reminding the physician to intervene. PMID- 11427246 TI - Rethinking practitioner roles in chronic illness: the specialist, primary care physician, and the practice nurse. AB - In this article, we describe an evidence-based stepped care approach to improving the care of chronic illness in organized health care systems. We review the common principles that have been found to improve the management and outcomes of patients with major depression, asthma, diabetes, and congestive heart failure. These population-based methods to improve care of chronic illness require reorganizing the roles of specialists, primary care physicians, and nurses. PMID- 11427247 TI - Effects of a modified group intervention with early-stage breast cancer patients. AB - The aim of this study was to investigate the clinical effectiveness of a psychiatric intervention program consisting of 5 weekly structured interventions and 3 additional group meetings every two months. Previous studies revealed that a 5 weekly structured intervention program was effective for alleviating psychological discomforts in Japanese breast cancer patients, and that the effectiveness persisted for 6 months for patients without lymph node metastasis or adjustment disorders. Since this 5-session intervention did not have persistent effects in patients with lymph node metastasis and/or adjustment disorders, 3 additional group meetings every two months were added after completion of the 5 weekly structured interventions. A total of 43 breast cancer patients completed the full program. The Profile of Mood States (POMS) scores were compared before, immediately after 5 sessions, immediately after the 3 additional interventions, and 6 months after all programs. As analyzed by POMS scores, the clinical effectiveness of a structured group intervention program persisted for 6 months for patients even with nodal metastases and/or adjustment disorders. These findings of the present study suggested that the 5 weekly intervention program was sufficient for patients without lymph node metastasis or adjustment disorders. In contrast, this intervention program alone was insufficient for patients with nodal metastases and/or adjustment disorders. For them, a new psychiatric intervention program consisting of 5 weekly structured interventions and 3 additional group meetings every two months were effective and sufficient. PMID- 11427248 TI - Referral pattern of neurological patients to psychiatric Consultation-Liaison Services in 33 European hospitals. AB - The referral pattern of neurological patients to 33 European psychiatric consultation-liaison (C-L) services in the general hospital was examined. Analyses were conducted on the ECLW CS data set, which consists of 14,717 psychiatric C-L referrals made in 56 European hospitals during 1991. Psychiatric referrals of patients admitted to neurological wards were compared to referrals from other wards. Information was obtained from 33 neurological wards, consisting of 34,506 neurological admissions. Of these admissions, 839 were referred to C-L psychiatry. The median consult rate among the hospitals was 1.8%. Compared to other hospital wards, patients referred from neurology were more frequently diagnosed as suffering from somatoform (P<.01) and dissociative disorders (P<.01), and less from substance abuse (P<.01) and delirium (P<.01). Referral to C-L psychiatry from neurological wards is characterized by an underestimation of psychiatric co-morbidity and a late detection, comparable to other medical specialties. An appeal is made for a standardized referral procedure including admission screening method, detecting patients at risk for nonstandard care during their hospital admission. PMID- 11427249 TI - Correlates of psychiatric morbidity in typhoid fever in a Nigerian general hospital setting. AB - This study explored factors associated with psychiatric morbidity in typhoid fever in a Nigerian general hospital. Information such as sociodemographic characteristics, symptom manifestations, results of investigations, neuropsychiatric symptoms, outcome and disposal were obtained from the case files of patients admitted for typhoid fever over a period of six years. The patients with psychiatric morbidity conspicuous enough to be documented by the attending physicians-mostly internists-were compared with those with no documented psychiatric morbidity on sociodemographic and clinical indices. Of the 136 cases, 26 (19.1%) had psychiatric morbidity. This included delirium (73.1%), generalized anxiety disorder (3.8%), depressive episode (3.8%), schizophrenia like disorder (3.8%) and monosymptomatic neuropychiatric manifestations such as apathy, hallucinations and irrelevant talking (15.5%). The clinical and sociodemographic indices that were significantly associated with psychiatric morbidity were diarrhea, blood biochemical imbalance and age (P<.05). Adolescents and young adults were more predisposed to developing psychiatric complications. Some factors potentially associated with psychiatric morbidity in typhoid fever have been identified. There is the need to prospectively assess the burden from psychiatric morbidity and identify interventions that may reduce it. PMID- 11427250 TI - Trichotillomania associated with dementia: a case report. AB - Trichotillomania represents a syndrome of hair pulling in which patients compulsively and ritualistically pluck their hair in response to a sense of tension or urgency. This report documents a case of hair pulling associated with dementia. Neuropsychological testing demonstrated the deficits to be localized predominantly to the frontal lobe dysfunction. PMID- 11427251 TI - Mutant hepatitis B viruses: a matter of academic interest only or a problem with far-reaching implications? PMID- 11427252 TI - MMR vaccine in HIV-infected children -- potential hazards? AB - Although given routinely as prophylaxis against wild-type measles to HIV-infected children, the live attenuated measles component of the MMR triple vaccine also possesses potential for disease. We document a case of measles proven to be caused by vaccine strain in a HIV-infected child here in the UK and discuss the clinical and immunological aspects. We also consider the new guidelines for MMR vaccination in HIV-infected children adopted last year in the USA. PMID- 11427253 TI - Modifying the cellular transport of DNA-based vaccines alters the immune response to hantavirus nucleocapsid protein. AB - Puumala virus is a member of the hantavirus genus (family Bunyaviridae) and is one of the causative agents of hemorrhagic fever with renal syndrome (HFRS) in Europe. A genetic vaccination approach was conducted to investigate if the immune response could be modulated using different cellular secretion and/or localisation signals, and the immune responses were analysed in BALB/c mice and in a bank vole infectious model. Rodents vaccinated with DNA constructs encoding the antigen fused to an amino-terminal secretion signal raised significantly higher antibody levels when compared to using constructs lacking secretion signals. Furthermore, the ratios of the IgG subclasses (IgG2a/IgG1) were raised by the use of cellular localisation signals, indicating a more pronounced Th1 type of immune response. The majority of the mice, or bank voles, immunised with DNA encoding a secreted form of the antigen showed a positive lymphoproliferative response and were protected against challenge with Puumala virus (strain Kazan wt). PMID- 11427254 TI - Nucleotide sequence analysis of varicella-zoster virus glycoprotein E epitope coding regions. AB - Varicella-zoster virus (VZV) glycoprotein E [gE] contains 623 amino acid residues. Fifty percent of the gE gene, codons 39 to 344 that encompasses two epitope coding regions e1 and c1, was sequenced and analyzed for variation among the 30 VZV isolates. A total of eleven isolates showed variance when compared with Dumas VZV strain sequence through base substitutions, with two isolates showing an amino acid change of tryptophan to arginine outside the coding regions of the epitopes e1 and c1 that are recognized by monoclonal antibodies 4F9 and c1, respectively. The results suggest that these epitopes were stable in the various VZV isolates. Thus, VZV glycoproteins with conserved epitopes are suitable candidates for both primary and booster vaccines. PMID- 11427255 TI - Experimental utility of rabies virus-neutralizing human monoclonal antibodies in post-exposure prophylaxis. AB - Rabies immune globulin (RIG) is essential for post-exposure prophylaxis but is expensive and not widely available. Rabies virus-neutralizing human monoclonal antibodies (Mabs) were evaluated in vitro and in a Syrian hamster model as a potential future alternative. Seven Mabs neutralized representative rabies virus variants. However, a European bat lyssavirus was not neutralized by either Mabs or RIG. Moreover, Duvenhage virus was neutralized by RIG, but not by Mabs, and Lagos bat and Mokola viruses were neutralized by one Mab but not by RIG. In hamsters, one Mab resulted in protection that was comparable to human RIG. These results suggest that Mabs may provide a promising alternative to RIG. PMID- 11427256 TI - Assembly of synthetic peptide vaccines by chemoselective ligation of epitopes: influence of different chemical linkages and epitope orientations on biological activity. AB - In this paper, we describe the assembly of synthetic peptide vaccines composed of a T helper cell epitope and a B cell epitope that were synthesized separately and then attached using three different chemoselective ligation methods: oxime bond formation, thioether bond formation and disulfide bond formation. The resulting vaccines were tested in animals to investigate their efficacy. We found that thioether bond formation gave the highest yield of material and that the chemistry involved did not adversely affect immunogenicity and biological activity of the peptide vaccine. Ligation of epitopes by oxime bond formation did not diminish biological activity either, but the yields of peptide vaccine were lower than when thioether bond formation was used. The vaccines in which a disulfide bond was used to attach the two epitopes resulted in the lowest yield and produced vaccines that also generated a weaker immune response with sub optimal biological activity. Connecting the T helper epitope via its N-terminus or its C-terminus to the N-terminus of the B cell epitope had little influence on resulting immunogenicity and biological activity. PMID- 11427257 TI - Tetanus toxoid loaded 'preformed microspheres' of cross-linked dextran. AB - The conformation of an antigen and hence its biological activity may get compromised when encapsulated in controlled release microspheres during formulation. In order to obviate the need for exposure of the antigen to the inactivating conditions, such as exposure to organic solvent and the high shear stress of emulsification required for microencapsulation, an alternate strategy was employed. 'Pre-formed' microspheres (20--340 microm in size) made of cross linked dextran (Dex) were employed as matrix for conjugation of tetanus toxoid (TT) under aqueous conditions. The native immunoreactivity of TT was completely retained after conjugation, as checked by immunofluorescence and quantitative ELISA. Immunogenicity of Dex--TT conjugate was tested in rodents. No untoward mortality or adverse effects of immunization with the test material were observed on histopathology of the site of injection. A single immunization with the long acting depot formulation elicited anti-TT antibody response lasting for 1 year without any need of booster. The titres were comparable after 12 weeks with those obtained using the conventional alum adsorbed toxoid. PMID- 11427258 TI - A classical inactivated vaccine induces protection against caprine herpesvirus 1 infection in goats. AB - Eight goats were inoculated with an inactivated caprine herpesvirus 1 (CpHV-1) vaccine. Four unvaccinated goats were kept as controls. After 30 days, four of the eight goats were revaccinated. Fifty days after the first vaccination the animals were divided into two groups (A and B) of six goats each (two goats vaccinated twice, two goats vaccinated once and two goats unvaccinated). The two groups were challenged with CpHV-1 intranasally (group A) or intravaginally (group B). After CpHV-1 challenge, the four control goats showed the typical genital lesions, whereas all the vaccinated goats were fully protected against clinical signs. Protection against infection depended on the number of injections of vaccine and the route of challenge. PMID- 11427259 TI - Present anti-measles immunity in Jordan. AB - The immunity of adults and the prevalence of measles was determined in order to evaluate the adequacy of current measles vaccination policy in Jordan. A total of 307 sera, collected from adults aged 18--40 years, were tested for anti-measles antibodies by ELISA technique. The overall prevalence of anti-measles antibodies was 94.8% and there was no significant difference in the seropositivity rate between males (95.7%) and females (94%). Of the tested adults, 71% were vaccinated and 29% escaped vaccination. In Jordan, measles outbreaks occur periodically and predominantly attack children aged 5--14 years. Our data support introducing the compulsory MMR vaccine in the year 2000 to improve the vaccination coverage of measles; since the optional single dose vaccination has not interrupted the circulation of the measles virus. A second dose of MMR vaccine could be offered to Jordanians either at school entry or at the age of 11 -12 years, based on the outcome of the compulsory single dose of MMR vaccine. PMID- 11427260 TI - Recombinant hemagglutinin protein of rinderpest virus expressed in insect cells induces humoral and cell mediated immune responses in cattle. AB - Rinderpest virus causes a highly contagious and often fatal disease in domestic and wild ruminants. The surface glycoproteins, hemagglutinin (H) and fusion (F) proteins of this enveloped virus are known to confer protective immunity in cattle. We have reported the generation of a recombinant baculovirus expressing H protein and studied its protective properties in cattle. In this report, we demonstrate that the recombinant baculovirus encoded H protein expressed in insect cells gets incorporated into extracellular baculovirus. Single administration of low doses of purified recombinant extracellular virus with or without adjuvant induces virus neutralizing antibody responses and bovine leukocyte antigen (BoLA) class II restricted helper T cell responses in cattle. PMID- 11427261 TI - A recombinant Semliki Forest virus particle vaccine encoding the prME and NS1 proteins of louping ill virus is effective in a sheep challenge model. AB - This study has examined the efficacy following intramuscular administration of a recombinant Semliki Forest virus (rSFV) vaccine, encoding the prME and NS1 proteins of louping ill virus (LIV), in sheep. Administration of rSFV-LIV vaccine resulted in transient detection at the injection site and draining lymph node only and no dissemination to distal sites. In addition, the recombinant vaccine offered complete protection against subcutaneous challenge with LIV, and partial protection following intranasal administration of LIV. Protected animals had no pathological changes normally associated with LIV infection, and had developed high antibody titres. In contrast, the two animals not protected exhibited classical clinical signs and neuropathological lesions of LIV infection. These findings indicate that rSFV-based vaccines have the potential to be developed as effective prototype vaccines for LIV. PMID- 11427262 TI - Gene gun-mediate DNA vaccination against foot-and-mouth disease virus. AB - Foot-and-mouth disease (FMD) is one of the most dangerous diseases of cloven hoofed animals and is a constant threat in the Middle-East and other regions throughout the world despite intensive vaccination programs. In this work, we describe the ability of FMDV expression constructs to protect pigs from FMDV challenge when used as a vaccine. The construct consists of encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES), the entire P1 and 2A together with 3CD sequences, all in the same reading frame. Another plasmid that was tested, carries the serotype O1 (G) VP1, Asia1 VP1 and O1 (G) 3C. Between each of the genes the 3C cleavage sequences were inserted. All constructs carried the cytomegalo virus (CMV) promoter. Using immunofluorescent and immunoblot techniques, we could show the expression and processing of viral proteins. Following the application of FMDV expression constructs into pigs skin by 'Gene Gun', pigs were partially protected from FMDV challenge. PMID- 11427263 TI - Staphylococcus aureus capsular polysaccharide type 5 conjugate and whole cell vaccines stimulate antibody responses in cattle. AB - Dairy heifers were immunized subcutaneously with one of four different vaccines which contained preparations of Staphylococcus aureus capsular polysaccharide type 5 (CP5) and a mineral oil adjuvant, or received a placebo containing saline and adjuvant. The vaccine containing a CP5-human serum albumin conjugate (CP5 HSA) and the vaccine with formaldehyde inactivated whole cells expressing CP5, both elicited strong anti-CP5 antibody responses. After two injections three weeks apart and a third injection 10 months later, the mean level and duration of the anti-CP5 antibody response was significantly higher in the whole cell group. No differences were found between the two groups with regard to the relative proportion of IgG subclasses, and the antibody responses to the polysaccharide were composed of both the IgG1 and IgG2. Vaccines containing only free CP5 or CP5 mixed with HSA produced weak and transient humoral immune responses. Only animals vaccinated with the whole cell vaccine or the conjugate vaccine showed responses to CP5 in a lymphocyte proliferation assay conducted one year after the third vaccination. This study indicates that CP5 expressed on the surface of formaldehyde inactivated whole cells, emulsified in an oil adjuvant, gives a strong and long lasting immune response in cattle. The use of conjugation technology, although effective, might not be necessary in order to achieve an immune response against S. aureus CP5 in cattle. PMID- 11427264 TI - Immunogenicity and reactogenicity of acellular diphtheria/tetanus/pertussis vaccines given as a pre-school booster: effect of simultaneous administration of MMR. AB - Four acellular diphtheria/tetanus/pertussis (aDTP) vaccines were compared with two diphtheria/tetanus (DT) vaccines given as a pre-school booster to 1033 children aged 4 to < 6 years who had completed primary immunisation with DTP vaccine according to the UK 2, 3 and 4 month schedule; 71 children had received aDTP vaccine and the remaining 962 a whole cell DTP vaccine for primary immunisation. The effect of simultaneous administration of a second dose of MMR vaccine was evaluated in 374 (37%). Overall, there was little difference in the frequency of post-vaccination symptoms in DT and aDTP vaccinees, although local reactions occurred more quickly in the aDTP group. The concomitant administration of MMR had no effect on local reactions or fever within 10 days, or on the proportions requiring a doctor's visit in the 4--6 week post-vaccination period. Local reactions > or = 3 cm were higher on day 2 in children who had received aDTP for primary immunisation (erythema 32.4% vs. 17.4% for wDTP, P = 0.0012; swelling 28.2% vs. 15.5%, P = 0.0027). Pertussis antibody responses were consistent with the antigen content of the aDTP vaccines. All were more immunogenic with respect to PT -- the only pertussis antigen which by itself has been shown to be protective in clinical trials -- than a wDTP pre-school booster given in an earlier trial. MMR vaccine had no significant effect on antibody responses to either the pertussis or diphtheria and tetanus antigens. Diphtheria antibody responses in children who had received wDTP for primary immunisation were 2.8 times higher than in those who had received aDTP vaccine (P < 0.0001); they were also higher in children who had received a single dose of a Haemophilus influenzae type b vaccine containing CRM(197) conjugate after 12 months of age. For countries currently using DT vaccines as a pre-school booster, replacement with an aDTP vaccine is unlikely to have a perceptible effect on reactogenicity, at least in children given wDTP for primary immunisation, and would boost antibody levels to antigens known to be associated with protection. PMID- 11427265 TI - The effect of introduction of universal childhood hepatitis B immunization in South Africa on the prevalence of serologically negative hepatitis B virus infection and the selection of immune escape variants. AB - The effect of universal hepatitis B vaccination on the prevalence of serologically negative hepatitis B virus infection (HBV) and the emergence of HBsAg variants is unknown. We prospectively studied two different cohorts of 12- 24 month old children in South Africa. They consisted of the unvaccinated children (n=459) born before the introduction of universal vaccination and the vaccinated children (n=1213) between 1 and 2 years after the introduction of the vaccination program. The frequency of detecting HBV DNA by PCR was reduced from 6.5% in unvaccinated children to 0.3% in vaccinated children (P<0.00001). There were no unique amino acid substitutions within the major hydrophilic region of the S sequence in both pre- and post-vaccination samples. Universal childhood vaccination reduced the frequency of serologically negative HBV infection and did not necessarily lead to selection of escape variants. PMID- 11427266 TI - The first five years of universal hepatitis B vaccination in South Africa: evidence for elimination of HBsAg carriage in under 5-year-olds. AB - South Africa implemented a vaccine against hepatitis B virus (HBV) into the Expanded Programme on Immunisation (EPI) in April 1995. The HBV vaccine is given at 6, 10, and 14 weeks, in parallel with OPV, DTP and Hib vaccines. This study assessed the impact of universal childhood HBV vaccination programme in reducing HBsAg carriage, in the first five years (1995--1999) since its implementation. In parallel, we investigated the current burden of HBV infection in mothers of vaccinees and the adult general population. A total of 598 babies (mean age=23.3 months) who received 3 doses of 1.5 microg/0.5 ml Hepaccine-B (Cheil) were recruited from the Northern Province (one of the nine provinces in South Africa). HBsAg, anti-HBs, anti-HBc, HBeAg and anti-HBe were tested using the IMx or Axsym kits (Abbott Laboratories). PCR assays were performed following established protocols. The overall seroprotection rate (i.e. anti-HBs titre> or =10 mIU/ml) was 86.8% (519/598) in vaccinated babies, while 13.2% had anti-HBs levels<10 mIU/ml. Seroprotection rates and geometric mean titres (GMT) decreased significantly with increasing age, possibly reflecting waning anti-HBs titre over time. Total HBV exposure (positive for either HBsAg, anti-HBs, or anti-HBc) was 31.0% (58/187) in mothers of vaccinees and 40% (72/180) in the adult general population. HBsAg carrier rate was virtually similar in both groups (3.2% in mothers of vaccinees vs. 3.3% in the general population). Against this background, no vaccine failures resulting in HBsAg and HBV DNA positivity were seen in vaccinated babies, including 6 babies born to HBsAg positive carrier mothers (one carrier mother was positive for HBeAg and HBV DNA). However, 0.9% (5/582) babies, aged between 8--11 months, tested positive for anti-HBc, all of whom had anti-HBs titres>10 mIU/ml and were negative for HBV DNA. Anti-HBc positivity was probably maternal in origin, or may represent sub-clinical averted HBV infections. It can be concluded that the HBV vaccine is highly effective within the framework of the South African EPI and already shows a positive impact in the elimination of HBsAg carrier rate in children<5 years. PMID- 11427267 TI - Expression of Helicobacter pylori urease subunit B gene in Lactococcus lactis MG1363 and its use as a vaccine delivery system against H. pylori infection in mice. AB - The use of Lactococcus lactis as an antigen delivery vehicle for mucosal immunisation has been proposed. To determine whether L. lactis could effectively deliver Helicobacter pylori antigens to the immune system, a recombinant L. lactis expressing H. pylori urease subunit B (UreB) was constructed. Constitutive expression of UreB by a pTREX1 vector resulted in the intracellular accumulation of UreB to approximately 6.25% of soluble cellular protein. Five different oral regimens were used to vaccinate C57BL/6 mice and the immune response measured. One regimen, which consisted of four weekly doses of 10(10) bacteria, followed after an interval of approximately 4 weeks by three successive daily doses, was able to elicit a systemic antibody response to UreB in the mice, although subsequently, a similar regimen produced a significant antibody response in only one out of six mice. The other three regimes, in which mice were vaccinated with two or three sets of three consecutive daily doses of recombinant bacteria over 30 days, failed to elicit significant anti-UreB serum antibody responses. In three regimens, the immunised mice were then challenged by H. pylori strain SS1 and no protective effect was observed. These findings suggest that any adjuvant effects of L. lactis are unlikely to be sufficient to produce an effective immune response and to protect against H. pylori challenge, when used to deliver a weak immunogen, such as UreB. PMID- 11427268 TI - Low-dose intradermal administration of recombinant hepatitis B vaccine in children: 5-year follow-up study. AB - Several studies have documented the efficacy of low-dose intradermal administration of hepatitis B vaccine. However, little is known about the duration of protection provided by low-dose intradermal administration of hepatitis B vaccine. This study reports results from a 5-year follow up period of 200 healthy children (100 infants and 100 preschool children) immunized intradermally with 2 microg doses of recombinant hepatitis B vaccine (GenHevac B) at months 0,1, and 6. In the 8th week after the third vaccine dose, 97% of the children developed anti-HBs antibodies higher than or equal to 10 mlU ml(-1), and the antiHBs geometric mean titre (GMT) was 676 mlU ml(-1). In month 18 and year 5, the anti-HBs GMT decreased to approximately one-third (220 mlU ml(-1)) and one tenth (68 mlU ml(-1)) of the initial levels, respectively. However, 87% of the children had protective levels of anti-HBs (> or =10 mlU ml(-1)) after 5 years. Among 156 children followed for 5 years, none became positive for anti-HBc and/or HbsAg. Seven children who were seronegative after 5 years developed anti-HBs antibodies higher than 1000 mlU ml(-1) after an additional 10 microg intramuscular hepatitis B vaccine. Persistent immunologic memory over periods of 5 years or more is evident, the anamnestic antibody response to a booster dose of vaccine, even in these children who have lost antibody. We conclude that intradermal administration of 2 microg recombinant hepatitis B vaccine provides long-term protection against hepatitis B virus in infants and preschool children. PMID- 11427269 TI - A truncated HCV core protein elicits a potent immune response with a strong participation of cellular immunity components in mice. AB - The immunogenicity of a truncated HCV core protein (Co.120) was studied in BALB/c and C57BL/6 mice, given three intramuscular injections of antigen, adjuvanted with either aluminum hydroxide or Freund's adjuvant. A rapid antibody response was noted after the first dose, with both strains of mice eventually exhibiting comparable levels of anti-core IgG (titers >1:100000), with a mixed IgG1/IgG2a subclass response. Spleen cells from Co.120-immunized mice gave a significant specific proliferative response. IFN-gamma gene expression was also detected after an ex-vivo specific stimulation of spleen cells in all immunized mice. This response was independent of dose, H-2 genetic background or type of adjuvant. The results indicated that immunization with the Co.120 protein elicits a potent anti HCV humoral and cellular immune response. PMID- 11427270 TI - Prothymosin alpha enhances protective immune responses induced by oral DNA vaccination against pseudorabies delivered by Salmonella choleraesuis. AB - Previously, we showed that vaccination with the glycoprotein D (gD) gene of pseudorabies virus (PrV) delivered by Escherichia coli induced protective immune responses. In this study, we report that oral DNA vaccination with attenuated Salmonella choleraesuis carrying the PrV gD gene conferred protective immunity in mice against PrV. Moreover, co-delivery of the prothymosin alpha gene carried by S. choleraesuis enhanced the vaccine efficacy. Our results thus demonstrate for the first time, to our knowledge, the effectiveness of oral DNA vaccination using S. choleraesuis as a delivery vehicle and the potential usefulness of prothymosin alpha as a DNA vaccine adjuvant. PMID- 11427271 TI - Three double-blind, randomized trials evaluating the safety and tolerance of different formulations of the saponin adjuvant QS-21. AB - The effects of the adjuvant QS-21 in various formulations on immediate pain on injection after intramuscular injection were evaluated in three Phase I clinical trials in healthy adults. Each trial was designed as a double-blind, randomized, four-way or five-way cross-over study with each subject acting as his/her own control. In the first trial, four formulations designed to evaluate the effect of QS-21 or pH (over a range of 6--7.2) were evaluated: phosphate-buffered saline at pH 6.0 or 7.2, and 50 microg of QS-21 in phosphate-buffered saline at pH 6.0 or 7.2. Thirty-three volunteers received each of the four intramuscular injections in random order separated by approximately 1 week. The volunteers assessed the immediate injection pain from 0 to 10 (none to most pain). The data indicate that the presence of QS-21, but not pH, is associated with transient injection site pain. The second trial, which utilized the same design as the first trial, evaluated formulations of QS-21 in various excipients. Fifteen volunteers received phosphate-buffered saline, QS-21/PBS, QS-21/aluminum hydroxide, and QS 21/4 mg/ml of polysorbate 80. Polysorbate 80, but not aluminum hydroxide, reduced the mean pain score compared to QS-21/PBS. The third trial evaluated formulations of QS-21 in additional excipients. Fifteen volunteers received aluminum hydroxide (without QS-21), QS-21/PBS, QS-21/0.72% benzyl alcohol, QS-21/30 mg/ml of hydroxypropyl-beta-cyclodextrin, and QS-21/8-mg/ml of polysorbate 80. Benzyl alcohol, cyclodextrin, and the higher concentration of polysorbate 80 reduced the pain scores associated with QS-21. Hence, QS-21 is associated with injection pain in simple buffer formulations, but it is possible to improve the acceptability of QS-21-containing formulations through reformulation with certain excipients. PMID- 11427272 TI - An open study of subcutaneous administration of inactivated hepatitis A vaccine (VAQTA) in adults: safety, tolerability, and immunogenicity. AB - A number of patients in clinical practice would be candidates for hepatitis A vaccine administered subcutaneously (SC), including patients with inherited and acquired coagulopathies. To assess the safety, tolerability, and immunogenicity of VAQTA (Hepatitis A Vaccine, Inactivated, Merck and Co. Inc., West Point, PA) was administered SC to healthy adults. A total of 114 healthy adults received two doses of vaccine SC 24 weeks apart. No serious vaccine-related adverse experiences were reported. Four weeks after dose 1, the seropositivity rate (SPR) was 77.9% (CI, 69.1, 85.1%). The geometric mean titer (GMT) was 21.0 mIU/ml. Twenty-four weeks after dose 1 (just prior to dose 2) and 28 weeks after dose 1 (4 weeks following dose 2), the SPRs were 95.3% [corrected] and 100%, respectively; the GMTs were 153.2 and 1563.9 mIU/mL, respectively [corrected]. Although the kinetics of the immune response were slower when VAQTA was administered SC compared to intramuscular injection, SPRs and GMTs increased over time, indicating that the vaccine administered SC demonstrated immunogenicity. PMID- 11427273 TI - Optimization of DNA immunization against human cytomegalovirus. AB - The immune responses of mice injected with plasmids VR-gB and VR-gB Delta tm expressing the full-length membrane-anchored, or secreted forms of human cytomegalovirus (HCMV)-glycoprotein B (gB), respectively, and VR-pp65 expressing the HCMV-phosphoprotein 65 (pp65) were analyzed. Pretreatment of mice with the local anesthetic bupivacaine did not enhance antibody production, and IFN-alpha co-expressed with the immunizing plasmids induced a moderate increase in the antibody response. However, antibody response was higher in mice inoculated at three sites in the musculus quadriceps than in mice inoculated at one site with the same dose and in the same muscle. pVR-gB Delta tm induced significantly higher antibody titers than the construct expressing the membrane-anchored form of gB, and priming with pVR-gB Delta tm followed by boosting with the gB subunit resulted in high-titer antibody responses. Immunization with VR-pp65 induced dose dependent CTL responses in about 50% of the mice at a dose of 50 microg. Co expression of IFN-alpha did not affect the number of responding mice. These findings might be important for optimization of humoral and cellular immune responses to HCMV after DNA vaccination. PMID- 11427274 TI - Immune responses and protection in different strains of aged mice immunized intranasally with an adjuvant-combined influenza vaccine. AB - Immune responses and protection against influenza virus infection were compared between young (2 months) and aged (18 months) BALB/c, C3H and C57BL/6 (B6) mice after intranasal vaccination. The mice were immunized with 2.5 microg protein of A/PR/8/34 (PR8) (H1N1) virus vaccine containing a cholera toxin adjuvant. In both the young and aged BALB/c mice, high levels of PR8-specific antibody-forming cell (AFC) responses were induced in the nasal-associated lymphoid tissue (NALT) 7 days after immunization. Nasal wash IgA and serum IgG antibody (Ab) responses to the PR8 haemagglutinin (HA) 4 weeks after immunization were slightly higher in the young mice than in the aged mice. The young mice showed complete protection against challenge infection, while the aged mice showed only a partial protection. In the C3H mice, NALT-AFC, and IgA and IgG Ab responses were higher in the young mice than those in the aged mice in parallel with the more efficient protection in the young mice than in the aged mice. Both the young and aged B6 mice showed no NALT-AFC responses, scarce IgA and IgG Ab responses and no protection. In the BALB/c mice, IgG1 and IgG2a levels were significantly lower in the aged mice. On the other hand, in the C3H mice, only IgG2a level was significantly lower in the aged mice. Similar results were obtained in terms of immune responses and protection between the young and aged mice of three different strains of mice after intra-nasal immunization with 0.1 microg of PR8 vaccine containing the adjuvant, two-times at 4-week intervals. In the B6 mice, the immune response was improved by immunization with a higher dose of the adjuvant-combined vaccine. These results suggest that local Ab responses, as well as systemic Ab responses, are downregulated in aged mice, although the degree of the downregulation of immune responses differs from strain to strain. PMID- 11427275 TI - Immunization of mice with recombinant gp41 in a systemic prime/mucosal boost protocol induces HIV-1-specific serum IgG and secretory IgA antibodies. AB - We tested the immunogenicity in mice of a recombinant fusion protein (gp41HA) consisting of the ectodomain of the HIV-1(IIIB) envelope glycoprotein gp41 fused to a fragment of the influenza virus HA2 hemagglutinin protein. An intraperitoneal prime followed by intranasal or intragastric boosts with gp41HA induced high concentrations of serum IgG antibodies and fecal IgA antibodies that reacted with gp41 in HIV-1(IIIB) viral lysate and were cross-reactive with gp41 in HIV-1(MN) lysate. By indirect immunofluorescence, serum IgG and fecal IgA from immunized mice were also shown to recognize gp41 in acetone-fixed human peripheral blood mononuclear cells infected with either syncytium-inducing (SI) or non-syncytium-inducing (NSI) North American HIV-1 field isolates, but not uninfected cells. Thus, this recombinant antigen may be useful in prime/boost immunization protocols designed to induce systemic and mucosal antibodies that recognize multiple primary HIV-1 isolates. PMID- 11427276 TI - Enhancement of the immunity to foot-and-mouth disease virus by DNA priming and protein boosting immunization. AB - Subunit vaccination is effective in eliciting humoral responses to a variety of viral antigens, however, it has not generated persistent protective immunity to foot-and-mouth disease virus (FMDV). In this study, we observed that priming mice with a DNA plasmid encoding VP1 of the FMDV O/Taiwan/97 capsid protein followed by boosting with a VP1 peptide conjugate (P29-KLH) resulted in production of not only high titers of antibodies but also antibodies with FMDV neutralizing activities. Moreover, the mice immunized in this manner cleared the virus from their sera in FMDV challenge experiments. Mice subjected to DNA plasmid priming and P29-KLH protein boosting had relatively higher ratio of IgG2a/IgG1 than those primed and boosted with P29-KLH conjugate. Addition of an oligodeoxynucleotide (ODN) containing immunostimulatory cytosine-phosphate-guanosine (CpG) motifs to P29-KLH conjugate also induced a higher ratio of IgG2a/IgG1 and significantly higher titer of neutralizing antibodies. These results indicate that treating animals with DNA plasmids priming and FMDV antigen(s) boosting may elicit immunity to FMD and this immune response may be augmented by CpG ODN. PMID- 11427277 TI - Protection of rainbow trout against infectious hematopoietic necrosis virus four days after specific or semi-specific DNA vaccination. AB - A DNA vaccine against a fish rhabdovirus, infectious hematopoietic necrosis virus (IHNV), was shown to provide significant protection as soon as 4 d after intramuscular vaccination in 2 g rainbow trout (Oncorhynchus mykiss) held at 15 degrees C. Nearly complete protection was also observed at later time points (7, 14, and 28 d) using a standardized waterborne challenge model. In a test of the specificity of this early protection, immunization of rainbow trout with a DNA vaccine against another fish rhabdovirus, viral hemorrhagic septicemia virus, provided a significant level of cross-protection against IHNV challenge for a transient period of time, whereas a rabies virus DNA vaccine was not protective. This indication of distinct early and late protective mechanisms was not dependent on DNA vaccine doses from 0.1 to 2.5 microg. PMID- 11427278 TI - A model to assess the infection potential of jet injectors used in mass immunisation. AB - Jet injectors are needleless injectors that penetrate skin with high-pressure fluid. They have potential advantages over needles and syringes in mass immunisation programs, but concerns over their capacity to transfer blood-borne viruses have been a barrier to acceptance. Hepatitis B infection can transmit in 10 pl of blood; detection of such low volumes presents severe difficulties to such assessments. A model to assess jet injector safety was developed using injection of an inert buffer into calves and assaying the next injector discharge, representing the next dose of vaccine, for blood using a highly sensitive ELISA. Four injectors were tested: two with reusable heads and direct skin contact, one with single-use injector heads and one where the injector head discharged at a distance from the skin. All injectors tested transmitted significant (over 10 pl) volumes of blood; the volumes and frequency of contamination varied with injector. The source of the contamination was consistent with contamination by efflux of injected fluid and blood from the pressurised pocket in tissue that is formed during injection. This insight should inform the design of safe jet injectors. PMID- 11427279 TI - Protective efficacy against tuberculosis of ESAT-6 secreted by a live Salmonella typhimurium vaccine carrier strain and expressed by naked DNA. AB - We have constructed a recombinant (r) attenuated Salmonella typhimurium strain which secretes ESAT-6 of Mycobacterium tuberculosis via the hemolysin secretion system of E. coli. Additionally, we have ligated ESAT-6 to different commercially available mammalian expression systems for use as naked DNA vaccines. We studied protection against M. tuberculosis induced by vaccination with each of these constructs alone or in combination in mice. Vaccination with a single dose of r S. typhimurium secreting ESAT-6 reduced numbers of tubercle bacilli in the lungs throughout the course of infection. The combined prime-boost vaccination did not considerably enhance protection. PMID- 11427280 TI - BBG2Na an RSV subunit vaccine candidate intramuscularly injected to human confers protection against viral challenge after nasal immunization in mice. AB - Respiratory syncytial virus (RSV) is a major respiratory pathogen responsible for severe pulmonary disease. We have developed a parenterally administered vaccine, BBG2Na, which is currently in a phase III clinical trial. BBG2Na comprises residues 130--230 of RSV-A G protein (G2Na) fused to the BB carrier protein. In this study, we show that BBG2Na can be delivered by the nasal route and generates both mucosal and systemic antibody responses when co-administered with cholera toxin B or a newly described delivery system, zwittergent 3--14. We found that nasal BBG2Na administration protects against RSV challenge and does not induce lung immunopathology upon subsequent RSV challenge. PMID- 11427281 TI - Paradoxical effects of IL-12 in leishmaniasis in the presence and absence of vaccinating antigen. AB - Protective immunity against Leishmania major requires parasite-specific CD4+T helper cells, the development of which is promoted by interleukin 12 (IL-12). In this study we investigated the use of IL-12 DNA to enhance the protective immunity induced by prophylactic vaccination with the L. major Parasite Surface Antigen 2 (PSA-2) DNA. A plasmid was constructed in which the two murine IL-12 subunits p35 and p40 were secreted as a biologically active single chain cytokine. The immunomodulatory effects of this IL-12 DNA were examined by codelivery with PSA-2 DNA in susceptible BALB/c and resistant C3H/He mice and subsequent infection with L. major promastigotes. Surprisingly, administration of IL-12 DNA alone had a protective effect, while coadministration of IL-12 with PSA 2 DNA abrogated protection. This effect of IL-12 DNA was dose dependent and affected by the timing of administration in relation to PSA-2 DNA. The effect of IL-12 on protection was associated with a reduced number of INF-gamma-producing T cells early in infection. A further understanding of this paradoxical effect of IL-12 and possibly other cytokines on protective immunity may be important for their use as adjuvants for Leishmania DNA vaccines. PMID- 11427282 TI - The immune response to a DNA vaccine can be modulated by co-delivery of cytokine genes using a DNA prime-protein boost strategy. AB - A large-scale DNA vaccination trial was performed in sheep to investigate whether co-delivery of the cytokine genes IL-4, IL-5, IL-15, GM-CSF or IFN-gamma could modulate the immune response generated to an antigen, in a DNA prime-recombinant protein boost regime. Vaccination with the recombinant EG95 protein has been shown to induce protection in sheep from Echinococcus granulosus infection, the causative agent of hydatid disease. Here we demonstrate that vaccination with DNA encoding EG95 effectively primed the humoral response, as judged by high IgG anti EG95 titres detected one-week after a boost with the recombinant protein. However, by two weeks after protein-boost the titres in the control group had reached levels similar to the groups primed with EG95 DNA. Priming with two doses of DNA vaccine followed by boosting with recombinant protein induced a predominantly IgG1 response. In contrast, priming and boosting with the protein vaccine generated a strong IgG2 response. Co-delivery of the EG95 DNA vaccine with DNA encoding GM-CSF enhanced the antibody titre to EG95 while co-delivery of IFN-gamma or IL-4 encoding DNA appeared to reduce the ability of the DNA vaccine to prime an IgG antibody response. This study has demonstrated the efficacy of the co-delivery of cytokines to modulate immune responses generated in a DNA prime-protein boost strategy. PMID- 11427283 TI - The anti-idiotypic antibody to chlamydial glycolipid exoantigen (GLXA) protects mice against genital infection with a human biovar of Chlamydia trachomatis. AB - Despite more than three decades of anti-chlamydial vaccine research and improved vaccine strategies with new technologies, no vaccine candidate has protected against heterologous challenge, nor at more than one site of infection. The majority of experimental anti-chlamydial vaccines to date have targeted the chlamydial major outer membrane protein (MOMP). Many MOMP-directed vaccine candidates have been highly immunogenic, but have failed to protect against infectious challenge. We have extended our previous studies of a different anti chlamydial vaccine, a monoclonal anti-idiotypic antibody (anti-Id; mAb2) which is a molecular mimic of the chlamydial glycolipid exoantigen (GLXA). The present studies demonstrate that the mAb2 vaccine is protective in a murine genital infection model utilizing a human urogenital strain. After either mucosal (oral or intranasal) or systemic (subcutaneous) immunization with the poly (lactide) encapsulated-mAb2 to GLXA, C3H/HeJ mice were significantly protected against topical vaginal challenge with Chlamydia trachomatis (K serovar; UW-31). Reduced vaginal shedding of organism and genital tract inflammation were associated with GLXA-specific and/or anti-EB neutralizing serum antibody. Our results demonstrate that the anti-Id (mAb2) vaccine is protective against an additional human biovar of C. trachomatis in C3H/HeJ mice, which are allogeneic to the source of mAb2 (BALB/c). PMID- 11427284 TI - Intranasal immunisation with influenza-ISCOM induces strong mucosal as well as systemic antibody and cytotoxic T-lymphocyte responses. AB - Intranasal administration of vaccines is preferred for induction of mucosal immune responses. In this study, mice were immunised intranasally and subcutaneously with influenza-immuno stimulating complexes (influenza-ISCOM). The intranasal dose was 15-times the subcutaneous dose. All mice dosed with influenza ISCOMs survived challenge with live virus and comparable serum antibody and splenic cytotoxic T-lymphocyte responses were detected in both groups. Induction of mucosal IgA was significantly higher with intranasal immunisation and was comparable to responses induced with the heat labile enterotoxin of Escherichia coli as adjuvant. These findings demonstrate that intranasal administration of high dose influenza-ISCOM results in potent systemic and mucosal immune responses. PMID- 11427285 TI - Evaluation of the response to a booster dose of hepatitis B vaccine in previously immunized healthcare workers. AB - INTRODUCTION: Hepatitis B vaccination is recommended for all healthcare workers (HCW) at risk of exposure to infectious body fluids. However, the absolute duration of protection from immunization is unknown. The purpose of this randomized comparison trial was to determine how previously immunized HCW respond to different booster doses of hepatitis B vaccine. METHOD: Adult HCW (n=59) were classified by level of hepatitis B surface antigen (anti-HBs), either <10 milli International Units per milliliter (mIU/ml) or 10-50 mIU/ml. Participants were then randomized to receive a 2.5 or 10 microg dose of hepatitis B vaccine. Evaluation of anti-HBs levels were conducted 10 to 14 days, one month and one year postbooster. RESULTS AND DISCUSSION: All participants responded to the booster dose with increased anti-HBs levels. At 14 days, mean anti-HBs levels were significantly higher for those with higher levels at baseline (P=0.004) and those receiving the 10 microg dose (P=0.016). At one month, those with higher anti-HBs levels at baseline and those receiving the 10 microg dose were significantly higher (P<0.01 for both). At one year, the increase for the higher dose was no longer statistically significant when examined by itself (P=0.081); statistical significance (P=0.021) was achieved after adjusting for anti-HBs level at baseline. For all participants, the geometric mean anti-HBs level was 2618 mIU/ml at 14 days, 2175 mIU/ml at one month and 88.9 mIU/ml at one year. At all time points the increase in anti-HBs levels represented an increase over the geometric mean baseline level of anti-HBs (7.4 mIU/ml). Hepatitis B immunized adults responded to a booster dose of hepatitis B vaccine from 3 to 13 yr postvaccination series. Data support current recommendations that immunized HCW do not require periodic antibody testing or vaccine boosters. PMID- 11427288 TI - The biomechanics and motor control of gait in Parkinson disease. AB - Parkinson disease is a progressive neurological condition characterised by hypokinesia (reduced movement), akinesia (absent movement), tremor, rigidity and postural instability. These movement disorders are associated with a slow short stepped, shuffling gait pattern. Analysis of the biomechanics of gait in response to medication, visual cues, attentional strategies and neurosurgery provides insight into the nature of the motor control deficit in Parkinson disease and the efficacy of current therapeutic interventions. In this article we supplement a critical evaluation of the Parkinson disease gait literature with two case examples. The first case describes the kinematic gait response of an individual with Parkinson disease to visual cues in the "off" phase of the levodopa medication cycle. The second case investigates the biomechanics and motor control of turning during walking in a patient with Parkinson disease compared with elderly and young control subjects. The results are interpreted in light of the need for gait analysis to investigate complex functional walking tasks rather than confining assessment to straight line walking, which has been the trend to date. PMID- 11427289 TI - Spinal posture and prior loading history modulate compressive strength and type of failure in the spine: a biomechanical study using a porcine cervical spine model. AB - OBJECTIVE: The purpose of this study was to investigate the effect of posture and loading history on the compressive strength and site of failure in the spine. DESIGN: An in vitro experiment was performed using a porcine cervical model that provided a homogeneous population of young healthy spines. BACKGROUND: The distribution of stresses amongst the many load bearing tissues of the spine is altered throughout the day by posture and the history of loading, but it is not clear how this modulates tissue damage or the risk of injury. METHODS: 48 porcine cervical spines were harvested and dissected into motion segments containing two vertebrae and the intervening disc (C3/4 and C5/6). Compressive loads and rotational torques (flexion/extension) were applied so that the effects of four loading histories (hydrated, neutral dehydration, flexed dehydration, superhydrated) and two failure postures (neutral, flexed) could be examined. Levels of dehydration were based on those reported over the course of a day. Dissection techniques and X-rays were used to document tissue damage. RESULTS. Specimens had a lower yield point (43--63%) and ultimate compressive strength (23 -47%) when in a flexed posture than when in a neutral posture. When injured in a neutral posture, superhydrated specimens had a lower strength (22--29%) than dehydrated specimens. Loading history also modulated the site of failure. CONCLUSIONS: The spine may be more prone to injury early in the morning when the discs are at their greatest level of hydration and/or when they are in a fully flexed posture. PMID- 11427290 TI - Kinematics of rotational mobilisation of the lumbar spine. AB - OBJECTIVE: The purposes of this study were to measure the movements of the lumbar spine produced by rotational mobilisation, and to study the effects of different grades of mobilisation on the movements produced. DESIGN: Kinematics of rotational mobilisation was assessed with an electromagnetic tracking device. BACKGROUND: Rotational mobilisation is frequently used in the treatment of back pain, but there was no information on its mechanical effects. METHODS: Movements of the lumbar spine were measured in 14 healthy volunteers when they were subjected to grades I to IV left rotational mobilisation. RESULTS: In the starting positions, the spines were found to be flexed, axially rotated to the left and laterally bent to the right. As the mobilisation grade increased, the spine was axially rotated further into the range. Rotational mobilisation was found to produce oscillatory movements of the lumbar spine in all three anatomical planes. It produced axial rotation which was accompanied by lateral bending in the opposite direction and sagittal rotation. The mean frequency of the oscillatory movements was 1.4 Hz. The amplitude of the oscillations was small, and was found to be increased in grades II and III mobilisation. CONCLUSION: Rotational mobilisation may be able to restore lost movements of the lumbar spine in any of the three anatomical planes. PMID- 11427291 TI - A three-dimensional mathematical model of temporomandibular joint loading. AB - OBJECTIVE: A mathematical model of the temporomandibular joint was developed to study the magnitude and direction of the compressive loading experienced at the temporomandibular joint during clenching. DESIGN: The model was based on the principles of static equilibrium in three dimensions. BACKGROUND: Direct measurement of temporomandibular joint loading in humans is extremely difficult. Animal models have provided an alternative in the past. However, evidence suggests that primates are not the most accurate human analogues for temporomandibular joint studies. A mathematical model was used as an alternative to direct measurement. METHODS: The EMG activity of two masticatory muscles was combined with their cross-sectional areas to calculate the force exerted by each muscle. Experimentally determined forces were implemented into a quadratic programming model to solve for the compressive forces on the joint. Two objective functions were chosen and their ability to predict muscle and joint forces was evaluated. RESULTS: The maximum bite forces for normal men, normal women, and women with temporomandibular joint disorders were 300 N (SD 102 N), 210 N (SD 57.7 N), and 120 N (SD 77.1 N), respectively. The calculated joint force for normal males was 260 N (SD 84.1 N). Normal females and female temporomandibular joint disorder patients produced temporomandibular joint forces of 172 N (SD 37.5 N) and 152 N (SD 44.2 N), respectively. PMID- 11427292 TI - Electromyographic and kinematic analysis of therapeutic knee exercises under water. AB - OBJECTIVE: This study aimed to evaluate muscle function and kinematics during commonly used knee rehabilitation exercises performed in water. DESIGN: Maximal effort single extension and flexion trials in still water and repeated extension flexion trials in flowing water in barefoot condition were analysed from 18 healthy participants (8 men, 10 women). BACKGROUND: Despite the fact that water exercises are widely used, there are only few studies involving biomechanical and hydrodynamical analysis of aquatic exercises in rehabilitation. METHODS: Electromyography of the quadriceps (vastus medialis, vastus lateralis) and hamstring muscles (biceps femoris, semitendinosus) and angular velocities of the movements were recorded under water. RESULTS: In the repeated extension-flexion exercises the early reduction of agonist activity occurred concurrently with a high level of activity of the antagonists. In the single trial exercises the level of antagonistic activity was low throughout the range of motion, whereas the level of agonist activity was higher during the final phase of the range of motion as compared with the repeated exercises. Angular velocity patterns and values were similar between the two types of exercises. CONCLUSIONS: The present data demonstrated that the flowing properties of water modified the neuromuscular function of the quadriceps and hamstring muscles acting as agonists and antagonists in the knee flexion-extension exercises. PMID- 11427293 TI - The effects of motion on force control abilities. AB - OBJECTIVE: To investigate the effects of motion on submaximal force control abilities in the knee extensors. DESIGN: Analysis of a force control task in a quasi-experimental design. BACKGROUND: Measuring strength control rather than strength capacity may provide insights in therapeutic intervention. METHODS: Twenty younger and 19 older community dwelling healthy adult males were instructed to maintain knee extensor strength at two levels of strength (20% and 60% of the maximal voluntary contraction) within a bandwidth (+/-6.10 N m) with visual feedback in isometric and isokinetic force control conditions. The effects of motion, force level, and age on force control ability were investigated. RESULTS: The addition of motion to the force control task decreased performance for both groups at both force levels. Isokinetic force control, compared to the isometric, increased force variability at both force levels. In isokinetic force control, as the force level increased, performance decreased for both groups. For the 60% maximal voluntary contraction isokinetic force control task, the older adults, compared to the younger, had an increased bias, increased percent time in bandwidth, and decreased number of correctional instances. CONCLUSIONS: This study has demonstrated that in the presence of motion, healthy younger and older adults experienced a decreased ability to maintain submaximal forces and their ability decreased even further with an increase in force level. Age group differences became more apparent by adding motion and increasing the force level. Further, we have successfully adapted the methodology used to assess isometric force control abilities to isokinetic force control testing in the lower extremities. PMID- 11427294 TI - Normative data for passive ankle plantarflexion--dorsiflexion flexibility. AB - OBJECTIVE: To describe the normal distribution parameters for measures of passive ankle plantarflexion-dorsiflexion flexibility obtained from a large sample of able-bodied young adult subjects. DESIGN: Seven variables were assessed and descriptive analyses were conducted. BACKGROUND: While assessment of plantarflexion-dorsiflexion flexibility is an important component of a clinical examination of plantarflexion contracture, there is limited normative data available that can be used as a reference for clinical decision-making. METHODS: Data were collected from 300 able-bodied male and female subjects aged between 15 and 34 years. Both ankles were measured. Load-displacement curves were collected using a manually controlled instrumented footplate. Six variables were extracted from these curves: passive torque at zero and 10 deg, passive stiffness at zero and 10 deg, and two coefficients from an equation fitted to the curve (i.e., k and b). The seventh variable, passive dorsiflexion range of motion, was quantified using a clinical procedure. RESULTS: Flexibility variables did not differ between the left and right ankles, nor between the dominant and non dominant legs. All variables were normally distributed. These distributions can, therefore, be adequately described using their mean and standard deviation values. CONCLUSIONS: This study has substantially increased the available database on plantarflexion-dorsiflexion flexibility and forms the basis of norm referenced clinical tests. PMID- 11427295 TI - Mild diabetic neuropathy affects ankle motor function. AB - OBJECTIVE: To evaluate the effect of age and diabetic neuropathy on ankle motor function in the frontal plane in terms of rate of torque development and capability for balance recovery. DESIGN: Case control study. Six older women with diabetic neuropathy compared to six women without neuropathy, matched for age and presence of diabetes mellitus; and nine healthy young women. BACKGROUND: Neuropathy causes a distal impairment in lower extremity sensory function which increases fall risk. Impairments in ankle inversion/eversion proprioceptive thresholds have been identified, but the effect of neuropathy on ankle motor strength in the frontal plane is unknown. METHODS: Subjects' abilities to recover from a lateral lean (with center of gravity offset as percentage of foot width) while standing on one foot, and to rapidly generate inversion torque about the ankle, were quantified. RESULTS: All nine of the young, but only one of six older, control subjects recovered from a 10% lean (P=0.0052). Three of six older controls, but no neuropathy subject, recovered from a 5% lean (P=0.083). Neuropathy subjects demonstrated half the ankle rate of torque development [78.2 (50.8) N m/s; P=0.016] of the young and older controls [162.0 (54.6) and 152.7 (22.2) N m/s, respectively]. CONCLUSIONS: Diabetic neuropathy leads to a decrease in rapidly available ankle strength which impairs balance recovery among older women. Younger women demonstrate similar ankle strength but superior balance recovery compared to older women without neuropathy. PMID- 11427296 TI - Influence of bone mineral density, age, and strain rate on the failure mode of human Achilles tendons. AB - OBJECTIVE: To examine the influence of strain rate, bone mineral density, and age in determining the mode by which human Achilles tendons fail. DESIGN: Dual-energy X-ray absorptiometry and mechanical testing of excised Achilles tendon-calcaneus specimens. BACKGROUND: The Achilles tendon can fail by tendon rupture or bony avulsion. These injuries are caused by similar loading mechanisms and can present similar symptoms. It is important to understand when each mode of injury is likely to occur so that accurate diagnoses can be made and appropriate treatments selected. METHODS: Excised human Achilles tendons were loaded to failure at strain rates of 1% s(-1) and 10% s(-1) following dual-energy X-ray absorptiometry examination to determine bone mineral density near the tendon insertion. Calcaneal bone mineral density, donor age, and strain rate were compared between specimens that failed by avulsion and those that failed by tendon rupture. RESULTS: While strain rate was not observed to affect failure mode, the calcaneal bone mineral density of specimens that failed by avulsion was significantly lower than the bone mineral density of specimens that failed by tendon rupture (P=0.004). There was a significant decrease in bone mineral density with age (P=0.004), and the difference in age between the avulsed and ruptured specimens was close to statistical significance (P=0.058). For the avulsed specimens, there was a significant linear relationship between failure load and bone mineral density squared (P=0.002). Logistic regression indicated that the effect of age on failure mode is secondary to the primary effect of bone mineral density. CONCLUSIONS: The avulsions were primarily "premature" failures associated with low bone mineral density. Since bone mineral density decreases with age, older individuals are more likely to experience avulsions while younger individuals are more likely to experience tendon ruptures. PMID- 11427297 TI - Medial displacement calcaneal osteotomy reduces the excess forces in the medial longitudinal arch of the flat foot. AB - OBJECTIVE: The hypothesis tested was that the increased load on the medial arch of the flat foot can be reduced through a medial displacement calcaneal osteotomy. DESIGN: A three-dimensional, biomechanical, multisegment model was used in conjunction with experimental data from the literature. BACKGROUND: Biomechanical models have been used to study the plantar fascia, medial arch height, subtalar motion and distribution of forces in the foot. METHODS: Responses of a normal foot, a flat foot and a flat foot with a medial displacement calcaneal osteotomy to an applied load of 683 Newtons were analyzed, and the distribution of support among the metatarsal heads and moment about various joints were computed. RESULTS: Compared to the normal foot, our flat foot model shifts the distribution of support from the lateral to the medial side, decreasing support provided by the fifth metatarsal from 11% to 1% of the total load, increasing support provided by the first metatarsal from 12% to 22% and increasing the moment about the talo-navicular joint from 20 to 28 Newton-meters. A ten millimeter medial displacement calcaneal osteotomy shifts support back toward the lateral side, with 11% provided by the fifth metatarsal and 13% by the first metatarsal. The moment at the talo-navicular joint decreases to eighteen Newton-meters. CONCLUSION: Our analysis indicates that a ten millimeter medial displacement calcaneal osteotomy in a flat foot model decreases the load on the medial arch. PMID- 11427298 TI - Contact stress on polyethylene components of a new rotating hinge with a spherical contact surface. AB - OBJECTIVE: To assess the nonlinear contact stress of a new rotating hinge of our knee prosthesis at various rotation angles. DESIGN: The contact surface between the metal tibial bearing and the ultra-high-molecular weight polyethylene plate of a conventional rotating hinge is of cylindrical design. We have designed a new type of rotating hinge with a congruous spherical contact surface. BACKGROUND: The endoprosthesis for reconstruction of limb after wide resection of malignant tumor around knee usually incorporates a rotating hinge. Our new rotating hinge with a spherical contact surface incorporates the benefits of an increased contact surface and potentially increased rotational stability during axial loading. METHODS: We utilized the ABAQUS finite element program to assess the nonlinear contact stress of this new rotating hinge at rotation angles of 0 degrees, 4 degrees and 8 degrees, based on a contact force of about 2800 N. RESULTS: The results show that von Mises stress for the finite element model of the polyethylene component of this rotating hinge ranges from 4.90 x 10(-6) to 8.22 MPa at the aforementioned rotational angles. The von Mises stress is about 1.31--1.82 MPa on the major parts of the ultra-high-molecular weight polyethylene plate, including both flanks. There is a mild stress concentration on the outer edge of polyethylene component, especially at 4 degrees and 8 degrees of rotation. The maximum values of von Mises stress at the contact surface at 0 degrees, 4 degrees and 8 degrees of rotation are 5.92, 7.49 and 8.22 MPa, respectively. These contact stresses are within the safety range of the ultra high-molecular weight polyethylene (compressive yield strength, 14 MPa). CONCLUSIONS: This new rotating hinge has an evenly distributed contact stress during axial load because of congruous contact design. PMID- 11427300 TI - Expression of neuronal isoform of nitric oxide synthase in spinal neurons of neonatal rats after sciatic nerve transection. AB - Motoneuron death induced by sciatic nerve transection in neonatal rats has been related to induction of the neuronal isoform of nitric oxide synthase (nNOS), a diaphorase of which one of the cofactors is nicotinamide adenine dinucleotide phosphate (NADPH). We transected the sciatic nerve of neonatal rats (P2) and examined nNOS expression by immunostaining in neurons of the sciatic pool and of other spinal levels on the 5th day after surgery. No correspondence was observed between the surviving motoneurons and nNOS positive cells. The appearance and distribution of nNOS positive neurons at all spinal levels and laminae were similar to those of adult animals. These results are at variance with previous studies which showed correlation between motoneuron loss after axotomy and number of NADPH-diaphorase positive motoneurons after sciatic transection. PMID- 11427301 TI - Influence of age on auditory startle responses in humans. AB - The auditory startle reaction is considered a brainstem reflex in response to an unexpected loud stimulus. We investigated the influence of age on auditory startle responses (ASRs) in 54 adult healthy subjects separated in three age groups (below 30; 30 to 50; above 50 years). ASRs were elicited by auditory stimuli randomly presented through tubal insert phones. Reflex electromyographic activity was simultaneously recorded from eight facial, neck, and extremity muscles. ASR probability was lower in extremity muscles of younger versus older subjects, but did not differ among age groups in facial and neck muscles. Median ASR latencies were significantly shorter in all muscles of younger versus older subjects. Our data provide evidence of a significant influence of age on ASRs. The observed differences are likely due to age-specific variations of central processing in the brainstem centers involved in ASR generation. PMID- 11427302 TI - Is the re-closure of surgically induced open neural tube defect the repetition of primary neurulation? An experimental study with chick embryos. AB - It is well known that surgically induced open neural tube defects (ONTDs) tend to re-close in early embryonic stages. To investigate whether the process of re closure is a repetition of primary neurulation, the morphological changes of surgically induced ONTDs were chronologically examined in early chick embryos. Longitudinal incisions of 3-somite length were made in the neural tubes of chick embryos at Hamburger and Hamilton stage 18 or 19. About half of the embryos showed complete closure of the skin on surgical microscopic examination. The re closed neural tubes showed no remarkable differences from the controls on histological examination. The process of re-closure involved a zipper-like (ventral to dorsal) closure in the transverse sections of neural tubes. The results indicate that the re-closure of surgically induced ONTDs does not repeat the process of natural primary neurulation. PMID- 11427303 TI - Increased innervation of rat preganglionic sympathetic neurons by substance P containing nerve fibers in response to spinal cord injury. AB - Substance P (SP) is elevated in the intermediate zone caudal to a spinal cord lesion presumably due to sprouting of intraspinal and primary afferent axons. It is unclear, however, if axon terminals are in direct contact with preganglionic neurons located within the different autonomic subnuclei. Therefore, the innervation of preganglionic sympathetic neurons by SP was quantified using confocal imaging and morphometric image analysis. The number of SP-immunoreactive varicosities apposed to nitric oxide synthase-positive neurons significantly increased bilaterally in all sympathetic areas of segment T2 one week after low cervical hemisection at C6/7. Consequently, direct excitatory effects of SP on preganglionic neurons may play an important role in the dysregulation of arterial blood pressure observed in patients with spinal cord injury at the cervical or upper thoracic level. PMID- 11427304 TI - Claudin-1, claudin-2 and claudin-11 are present in tight junctions of choroid plexus epithelium of the mouse. AB - The choroid plexus epithelium forms the blood-cerebrospinal fluid (CSF) barrier and is responsible for the secretion of the CSF from the blood. The morphological correlate of the blood-CSF barrier are the tight junctions of choroid plexus epithelium. By freeze-fracture electron microscopy it has been demonstrated that choroid plexus epithelial tight junctions form parallel strands resembling those of Sertoli cells building the blood-testis barrier and those of the myelin sheaths of oligodendrocytes. As the oligodendrocyte specific protein/claudin-11 has been shown to be the central mediator of parallel-array tight junctions in Sertoli cells and myelin sheaths in mice, we asked whether claudin-11 is present in the tight junctions of choroid plexus epithelial cells of the mouse. Here, we present the first direct evidence that claudin-11 besides claudin-1 and -2, occludin and the zonula occludens protein ZO-1 is present in choroid plexus epithelial tight junctions. During inflammation in the central nervous system such as experimental autoimmune encephalomyelitis, the molecular composition of choroid plexus epithelial tight junctions does not change considerably. Their unique molecular composition, with claudin-11 accompanied by claudin-1 and claudin-2 points to a unique regulatory mechanism of the blood-CSF-barrier function. PMID- 11427305 TI - Loss of plasma glucose lowering response to cold stress in opioid mu-receptor knock-out diabetic mice. AB - Opioid mu-receptor plays an important role in the regulation of glucose homeostasis in diabetic rats lacking insulin. Opioid mu-receptor knockout mice were employed to identify the essential role of this receptor in the present study. Western blotting analysis characterized the deletion of opioid mu-receptor in liver of knockout mice as compared to that of normal (wild-type) mice. We found that the plasma glucose concentration of diabetic mice induced by intraperitoneal injection of streptozotocin was markedly decreased after exposure to cold-stress in a cold room for 1 h. However, this plasma glucose lowering response to cold-stress was disappeared in diabetic mice lacking opioid mu receptor. The important role of opioid mu-receptor in the plasma glucose lowering response to cold stress can thus be considered. Moreover, bilateral adrenalectomy abolished this plasma glucose lowering response to cold stress in diabetic mice with opioid mu-receptor, as compared to the shamed-operated animals. Therefore, activation of opioid mu-receptor by opioid from adrenal gland appears to be responsible for the plasma glucose lowering response to cold-stress in diabetic mice with insulin deficiency. PMID- 11427306 TI - Changes in Ca2+-dependent protein kinase C isoforms induced by chronic ethanol treatment in mice. AB - Protein kinase C (PKC) has been shown to regulate ethanol sensitivity. The goal of the present study was to ascertain whether chronic in vivo ethanol treatment could affect PKC isoforms in the mouse brain. We measured the protein level of membrane-bound PKC isoforms following chronic ethanol treatment using Western blotting. The protein level of membrane-bound PKCalpha and PKCgamma isoforms, which are defined as Ca2+-dependent PKC isoforms (cPKC), in the limbic forebrain during chronic ethanol treatment was significantly increased, whereas the levels of both were significantly decreased in the frontal cortex. By contrast, there was no change in PKCepsilon, a Ca2+-independent PKC isoform, in both areas. These findings suggest that the change in membrane-bound cPKC in the limbic forebrain and frontal cortex may play substantial roles for the development of ethanol dependence. PMID- 11427307 TI - Deactivation and desensitization of mouse embryonic- and adult-type nicotinic receptor channel currents. AB - Recombinant nicotinic acetylcholine receptor (nAChR) channels transiently expressed in HEK293 cells were investigated using the patch-clamp technique in the cell-attached and outside-out modes for single-channel analysis and ultra fast agonist application to multiple channels. Deactivation (current decay after removal of agonist) and desensitization (current decay in the presence of agonist) were analyzed at embryonic- (gamma) and adult-type (epsilon) nAChR channels. Time constants of desensitization were similar for both receptor types (epsilon: 53.1+/-16.9 ms; gamma: 49.2+/-15.7 ms) and corresponded to the mean duration of clusters of single channel openings activated by pulses of 1 mM ACh. Deactivation showed distinct characteristics. Time constants were 1.76+/-0.16 ms for epsilon- and 3.19+/-0.18 ms for gamma-nAChR channels, corresponding to mean burst duration analyzed from single channels in the same preparation (epsilon: 1.85+/-1.2 ms, gamma: 3.85+/-2.1 ms). It is assumed that differences in deactivation are of functional relevance at the muscle endplate. PMID- 11427308 TI - Prepubertal increases in the levels of two salmon gonadotropin-releasing hormone mRNAs in the ventral telencephalon and preoptic area of masu salmon. AB - Ontogenic changes in the expression levels of two salmon gonadotropin-releasing hormone genes (sGnRH-I and -II) were examined in the forebrain region including the ventral telencephalon and preoptic area of masu salmon by competitive reverse transcription-polymerase chain reaction (RT-PCR). Two genes showed similar expression patterns throughout the lifetime in both sexes, although the levels of sGnRH-II mRNA were about 20 times higher than those of sGnRH-I mRNA. In males, the levels of sGnRH mRNAs increased at the beginning of the second year and reached their maximum in the autumn. The levels decreased gradually until the autumn of the third year when fish sexually matured. In females, the levels reached their maximum in the first autumn and fluctuated considerably along with the seasons in the third year. These results suggest that, in the salmon brain, sGnRH genes are activated long before the sexual maturation under sexually different control mechanisms. PMID- 11427309 TI - Isoflurane slows inactivation kinetics of rat recombinant alpha1beta2gamma2L GABA(A) receptors: enhancement of GABAergic transmission despite an open-channel block. AB - Recombinant alpha1beta2gamma2L gamma-aminobutyric acid (A) receptor (GABA(A)R) channels expressed in human embryonic kidney (HEK293) cells were used for patch clamp experiments. The currents activated by brief pulses of GABA (10(-4) M) applied with a device for fast solution exchange to cells clamped in the whole cell configuration mimicked GABA(A)R-mediated inhibitory postsynaptic currents. Isoflurane (ISO) at clinically relevant concentrations (0.6 mM) decreased the amplitude and prolonged the decay of the GABA-evoked response. To further detail the mechanism underlying the prolonged decay time, we made simulations based on these measurements. These simulations suggest that ISO slows the rate of GABA unbinding from the receptor. Under these conditions, ISO increases the GABA induced charge transfer and, thus, could enhance GABAergic inhibition despite the concomitant open-channel block causing the decrease in the current amplitude. PMID- 11427310 TI - Characterization of murine immunoglobulin G antibodies against human amyloid beta1-42. AB - It has been demonstrated that immunization of transgenic mouse models of Alzheimer's disease (AD) with amyloid-beta1-42 peptide (Abeta1-42) results in amelioration of AD-like pathology, including reduced soluble and deposited beta amyloid and decreased cognitive impairment. Based on the proposed importance of immunoglobulin G (IgG) anti-Abeta antibodies (Abs) in these effects, we sought to characterize these Abs in splenocytes from mice immunized with Abeta1-42. Data show that a more aggregated preparation of Abeta1-42 gives a robust IgG anti Abeta Ab response, while these Abs are almost undetectable when a less aggregated preparation of Abeta1-42 is used as the immunogen. Importantly, IgG anti-Abeta Ab production is detected after just 12 weeks of Abeta1-42 treatment. Analysis of anti-Abeta Ab IgG isotypes reveals that the majority of these Abs are IgG1, with significantly fewer Abs of the IgG2a or IgG2b isotypes (IgG1>IgG2a>IgG2b), suggesting a T lymphocyte helper type II response after Abeta1-42 immunization. To determine the epitope of Abeta recognized by IgG anti-Abeta Abs, intact Abeta and Abeta peptide fragments were analyzed for their ability to bind these Abs. Data show that these Abs specifically recognize an amino-terminal epitope of Abeta between amino acids one and twelve, with higher affinity for a more soluble preparation of Abeta1-42. These data further indicate the immunogenic potential of Abeta1-42 and offer insight into the nature of the IgG anti-Abeta Ab response. PMID- 11427311 TI - Activation of the hypothalamus characterizes the acupuncture stimulation at the analgesic point in human: a positron emission tomography study. AB - We performed a positron emission tomography study, using regional cerebral blood flow as the index of brain activity, to address the specificity of brain activation pattern by acupuncture stimulation of short duration at the classical analgesic point. Needling manipulation at 2 Hz was performed at a classical point of prominent analgesic efficacy (Li 4, Heku) and a near-by non-classical/non analgesic point, respectively, in normal subjects. Regions activated by acupuncture stimulation at Li 4 included the hypothalamus with an extension to midbrain, the insula, the anterior cingulate cortex, and the cerebellum. Of note, it was only the stimulation at Li 4 that activated the hypothalamus under the similar psychophysical ratings of acupuncture sensation (deqi) as elicited by the stimulation at the two points, respectively. The data suggested that the hypothalamus might characterize the central expression of acupuncture stimulation at the classical analgesic point and serve as one key element in mediating analgesic efficacy of acupuncture stimulation. PMID- 11427312 TI - Increased excitability in the primary motor cortex and supplementary motor area in patients with phantom limb pain after upper limb amputation. AB - Using functional magnetic resonance imaging and single slice FLASH technique, we investigated reorganization of the hand representation of the primary sensorimotor cortex (SMC) in 16 patients with upper extremity amputation. Patients were asked to perform finger tapping with the intact hand, repetitive eye closing and anteflexion of the amputation stump or intact shoulder. Six normal volunteers served as control. In the normal volunteers activations during shoulder anteflexion, finger tapping and eye closure were located within the central sulcus in a medio-lateral fashion. Patients demonstrated invasion of the face or shoulder representation into the hand representation of the amputated limb. Eight phantom limb pain patients showed significantly greater activation in SMC and supplementary motor area (SMA) in contrast to eight patients without phantom limb pain. We conclude, that different parts of the motor system are affected in patients with phantom limb pain--possibly in the sense of an up regulation of excitability. PMID- 11427313 TI - Auricular acupuncture decreases neuropeptide Y expression in the hypothalamus of food-deprived Sprague-Dawley rats. AB - The aim of the present study was to investigate whether stimulation of auricular acupuncture point has any effects on the expression of neuropeptide Y (NPY), appetite-inducing factor particularly abundant in the mammalian hypothalamus. In food-deprived condition, enhanced NPY expression was detected in both the arcuate nucleus (ARN) and the paraventricular nucleus (PVN) of the hypothalamus via immunohistochemistry in Sprague-Dawley rats. Needling the unfed rats on the auricular point resulted in decreased NPY levels in both the ARN and PVN, while it increased NPY levels in the ARN and PVN of fed rats. The present findings indicate that auricular acupuncture may affect NPY expression in the ARN and PVN of the hypothalamus. PMID- 11427314 TI - Characterization of 5' flanking region of alpha isoform of rat Ca2+/calmodulin dependent protein kinase II gene and neuronal cell type specific promoter activity. AB - The 5' flanking region of the alpha isoform of the rat Ca2+/calmodulin-dependent protein kinase II (alpha CaM kinase II) gene was isolated in 2.3 kbp of genomic sequence. Functional analysis of alpha CaM kinase II promoter deletion mutants fused to a reporter gene in neuroblastoma, including N18TG2, NG108-15, and CAD cells revealed strong transcriptional activity localized 100-145 bp, and a potent silencer 199-275 bp upstream of the transcription start site. The promoter is inactive in non-neuronal cells including BALB/c 3T3, Chinese hamster ovary, HT1080, and C6 glioma cells. These results indicated that the alpha CaM kinase II gene is transcribed from a tissue-specific promoter which is under intense negative control. PMID- 11427315 TI - Reduced cerebrospinal fluid estradiol levels are associated with increased beta amyloid levels in female patients with Alzheimer's disease. AB - Recent in-vitro studies indicate that estrogens such as 17beta-estradiol (E2) may decrease the production of beta-amyloid 1-42 (Abeta42), a peptide central for the formation of senile plaques in Alzheimer's disease (AD). To test this hypothesis in a clinical study, cerebrospinal fluid levels of E2 were compared between 30 female AD patients and 11 female patients with non-dementing diseases such as major depression and investigated with respect to beta-amyloid 1-40 and Abeta42 levels. E2 levels were significantly (P<0.05) lower in the AD group than in controls; within the AD group E2 levels were inversely correlated with Abeta42 concentrations (r=-0.36, P=0.05). This is the first clinical study providing evidence for an influence of E2 on Abeta42 metabolism in vivo. This observation corresponds to the putative beneficial effects of estrogen replacement therapy on the development and course of AD. PMID- 11427316 TI - No pathogenic mutations in the synphilin-1 gene in Parkinson's disease. AB - alpha-Synuclein is mutated in rare autosomal dominant forms of Parkinson's disease and is a major component of Lewy bodies and neurites. Synphilin-1, a novel protein interacts in vivo and co-localises with alpha-synuclein in Lewy bodies. We analysed the synphilin-1 gene in familial Parkinson's disease by single-strand conformation polymorphism (SSCP) and automated sequencing but found no coding mutations. However, we identified two novel intronic polymorphisms; an A/T polymorphism in intron 2, resulting in the introduction of an Alu1 site and a second G/T polymorphism in intron 4. We analysed the intron 2 polymorphism for allelic association as it was conducive to rapid screening but observed no changes in frequency between Parkinson's disease cases and controls. PMID- 11427317 TI - Influence of interleukin-1beta gene polymorphism on age-at-onset of spinocerebellar ataxia 6 (SCA6) in Japanese patients. AB - An inverse correlation is observed between the expanded CAG repeat number and age at-onset of spinocerebellar ataxia 6 (SCA6). To detect another modifying genetic factor for SCA6, we studied polymorphisms in the genes for interleukin (IL)-1beta and tumor necrosis factor in 122 Japanese patients with SCA6. No contribution of these polymorphisms to the variance in disease onset was observed by regression analysis or by ANOVA. The IL-1beta promoter polymorphism, however, significantly affected the age-at-onset, when adjusted for the CAG repeat number as a covariate (P=0.0004, by ANCOVA), suggesting that IL-1beta may be a genetic factor other than the SCA6 gene that modifies the age-at-onset of the disease. PMID- 11427318 TI - Detection of brain-specific autoantibodies to myelin oligodendrocyte glycoprotein, S100beta and myelin basic protein in patients with Devic's neuromyelitis optica. AB - Neuromyelitis optica (NMO) is a rare syndrome characterized by the combination of acute optic neuritis and transverse myelitis, usually not seen in Multiple Sclerosis (MS) and other demyelinating syndromes of the central nervous system (CNS). A high prevalence of various autoantibodies has been described in patients with NMO suggesting a polyclonal activation of the humoral immune system. We examined autoantibody responses to myelin (MBP, MOG with isotypes and epitopes) and astroglial (S100beta) antigens in four patients with NMO by ELISA and Immunoblot. All patients showed a positive anti-MOG response, with one showing reaction to the MOG epitope corresponding to amino acid sequence 63-87. MBP autoantibodies were only detected in two and S100beta-autoantibodies in one patient. Despite the limited number of samples, these findings suggest a predominant anti-MOG rather than anti-MBP or anti-S100beta autoantibody response in NMO, though no NMO-specific antibody pattern was found, which is in keeping with a widespread acute immune activation, including a strong B-cell response. PMID- 11427319 TI - Chest pain in the absence of coronary artery disease: a biopsychosocial perspective. PMID- 11427320 TI - Mapping a gene for neuropathic pain-related behavior following peripheral neurectomy in the mouse. AB - Total hindpaw denervation in rodents elicits an abnormal behavior of licking, scratching and self-injury of the anesthetic limb ("autotomy"). Since the same denervation produces phantom limb pain and anesthesia dolorosa in humans, autotomy has been used as a model of human neuropathic pain. Autotomy is an inherited trait in rodents, attributable to a few genes of major effect. Here we used recombinant inbred (RI) mouse lines of the AXB-BXA RI set to map a gene for autotomy. Autotomy levels following unilateral sciatic and saphenous nerve section were scored daily for 36 days, using a standardized scale, in all 23 RI lines available for this set. We used a genetic map of 395 marker loci and a permutation-based statistical method for categorical data to assess the statistical significance of mapping results. We identified a marker on chromosome 15 with statistical support (P=0.0003) in the range considered significant for genome-wide scans in the mouse. Several genes located in this chromosomal region encode for neural functions related to neuropathic pain and may indicate targets for development of novel analgesics. PMID- 11427321 TI - Osteoarthritis and its association with muscle hyperalgesia: an experimental controlled study. AB - Hypertonic saline effectively excites muscle nociceptors. Muscle hyperalgesia was assessed in osteoarthritis (OA) by intramuscular infusion of 0.5 ml hypertonic saline (6%) into the tibialis anterior muscle in humans. Patients (n=14) with OA in the lower extremities were compared with an equal number of age- and sex matched healthy controls. Ten of the 14 OA patients had pain in the knee joint as the most common presenting complaint. Visual analogue scale (VAS) pain intensity and assessment of pain areas were recorded before infusion and immediately, 2, 5, 10 and 20 min after infusion, and then every 10 min, until the pain vanished. The mean pain offset time in OA patients (11.3+/-7.9 min) was larger as compared with the control subjects (6.04+/-2.1 min) (P=0.025). OA patients had increased pain intensity VAS after the infusion in the right leg compared with controls (P<0.05). Referred and radiating pain areas at 2 min post-infusion increased in OA patients and not in controls as compared with the local pain areas (P<0.05). It is concluded that muscle hyperalgesia and extended pain areas might be due to central sensitization caused by painful osteoarthritis. PMID- 11427322 TI - Comorbidity of pain-associated disability and depressive symptoms in connection with sociodemographic variables: results from a cross-sectional epidemiological survey in Hungary. AB - The purpose of this study was to investigate the prevalence of pain symptoms causing disabilities in every-day activities and their possible connection to depressive symptomatology. A representative sample of 12640 adults from the Hungarian population participated in a door-to-door survey about demographic variables, pain-associated disability, and depressive symptomatology. The overall prevalence of pain-associated disability was 32.7%, significantly lower in men, showing a significant increasing trend with age. A decreasing tendency in prevalence rates was observed in connection with higher educational and occupational status. Results revealed a 30.2% prevalence of depressive symptomatology among interviewees reporting pain-associated disabilities. The co prevalence of depressive symptoms revealed a significant increasing trend with age and lower educational level. No significant gender difference was found in the co-occurrence of depressive symptoms. This survey concludes that pain symptoms constitute a substantial public health problem in the Hungarian population in forms of emerging disabilities and depression. Epidemiological studies offer a better understanding of sociodemographic differences in health status, and serve the better allocation of professional and economic resources. PMID- 11427323 TI - Reduced pain tolerance during and after pregnancy in women suffering from fear of labor. AB - Pain tolerance in women suffering and not suffering from fear of labor during and after pregnancy were compared. Twenty women with labor fear and 20 control women were subjected to a cold pressor test (CPT) on average 1 month before delivery and 9 months later. Half the volunteers were nulliparous and half parous. Pain endurance time (PET) and intensity of pain (Visual Analogue Scale, VAS) during CPT was assessed. Patients in the fear group tolerated CPT for a significantly shorter time than did women without fear both in pregnancy (154.8+/-109.9 s vs. 282.5+/-60.1 s (mean+/-SD), P<0.001), and in the postpartum period (128.6+/-111.7 s vs. 279.6 +/-60.3 s, P=0.002). Those with labor fear experienced CPT as significantly more painful than did the controls both during pregnancy (VAS 6.68+/-1.9 vs. 3.78+/-2.0 (mean+/-SD), P<0.001) and after (VAS 7.73+/-1.5 vs. 5.92+/-3.0, P=0.04). PET and VAS values correlated during pregnancy (r=-0.62, P<0.001), but not after pregnancy (r=-0.30, not significant). Parity was not associated with either PET or VAS scores. As a rule, pain in all women during CPT was regarded to be lower during pregnancy than after pregnancy (VAS 4.87+/-2.4 vs. 6.60+/-2.6, P=0.001). Patients with fear of labor were characterized by pain intolerance also in circumstances other than labor. This fact may indicate enhanced sensitivity to pain-causing mechanisms in women who develop fear of labor. PMID- 11427324 TI - Working with low back pain: problem-solving orientation and function. AB - A number of ergonomic, workplace and individual psychosocial factors and health behaviors have been associated with the onset, exacerbation and/or maintenance of low back pain (LBP). The functional impact of these factors may be influenced by how a worker approaches problems in general. The present study was conducted to determine whether problem-solving orientation was associated with physical and mental health outcomes in fully employed workers (soldiers) reporting a history of LBP in the past year. The sample consisted of 475 soldiers (446 male, 29 female; mean age 24.5 years) who worked in jobs identified as high risk for LBP related disability and reported LBP symptoms in the past 12 months. The Social Problem-Solving Inventory and the Standard Form-12 (SF-12) were completed by all subjects. Hierarchical multiple regression analyses were used to predict the SF 12 physical health summary scale from interactions of LBP symptoms with each of five problem-solving subscales. Low scores on positive problem-solving orientation (F(1,457)=4.49), and high scores on impulsivity/carelessness (F(1,457)=9.11) were associated with a steeper gradient in functional loss related to LBP. Among those with a longer history of low-grade LBP, an avoidant approach to problem-solving was also associated with a steeper gradient of functional loss (three-way interaction; F(1,458)=4.58). These results suggest that the prolonged impact of LBP on daily function may be reduced by assisting affected workers to conceptualize LBP as a problem that can be overcome and using strategies that promote taking an active role in reducing risks for LBP. Secondary prevention efforts may be improved by addressing these factors. PMID- 11427325 TI - Cognitive, behavioral and physiological reactivity to pain as a predictor of long term pain in rheumatoid arthritis patients. AB - A heightened reactivity to pain is assumed to play a significant role in the maintenance and exacerbation of pain in patients with chronic pain. In a prospective study involving 95 rheumatoid arthritis (RA) patients, the relative contribution of self-reported cognitive, behavioral and physiological components of pain reactivity were examined for a change in pain within 1 year. Regression analyses indicated that self-reported physiological reactivity predicted an increase in clinical and self-reported pain after 1 year, but not cognitive and behavioral reactivity. Neither disease activity nor neuroticism mediated or moderated the relationship of pain reactivity to long-term pain. However, structural equation modeling revealed that neuroticism directly affected physiological reactivity to pain, which in turn was the only significant predictor for subsequent pain. The results of this study underline the crucial role of physiological pain reactivity for exacerbation of pain in RA patients and are indicative for a symptom-specific pattern of physiological pain reactivity that is sustained by psychological predisposition and respondent learning processes. PMID- 11427326 TI - Effect of gabapentin and lamotrigine on mechanical allodynia-like behaviour in a rat model of trigeminal neuropathic pain. AB - Injury to the trigeminal nervous system may induce severe pain states. This study examined the antinociceptive effect of the novel anticonvulsants, gabapentin and lamotrigine, in a rat model of trigeminal neuropathic pain produced by chronic constriction of one infraorbital nerve. Responsiveness to von Frey filament stimulation of the vibrissal pad was evaluated 2 weeks post-operation. Hyper responsive rats received acute and repeated (five injections separated by the half-life of the compound) injections with gabapentin and lamotrigine. 76% of the nerve-injured rats displayed pronounced hyper-responsiveness (median 0.217 g (lower-upper percentiles 0.217-0.217) vs. 12.5 g pre-operative), that was resistant to both single (5-100 mg/kg) and repeated (5-30 mg/kg) injections with i.p. lamotrigine. Repeated (30 and 50 mg/kg), but not single (30-100 mg/kg) injections of i.p. gabapentin partially alleviated the mechanical allodynia-like behaviour. Repeated injections of gabapentin at 50 but not at 30 mg/kg produced motor deficits. The results indicate that gabapentin rather than lamotrigine may be a better therapeutic approach for the clinical management of some trigeminal neuropathic pain disorders. PMID- 11427327 TI - Methadone maintenance patients are cross-tolerant to the antinociceptive effects of morphine. AB - We have previously shown that methadone maintenance patients are hyperalgesic. Very little is known about the antinociceptive effects of additional opioids in these patients. This study (1) compared the intensity and duration of antinociceptive responses, at two pseudo-steady-state plasma morphine concentrations (C(SS1) and C(SS2)), between four patients on stable, once daily, doses of methadone and four matched control subjects; and (2) determined, in methadone patients, whether the antinociceptive effects of morphine are affected by changes in plasma R(-)-methadone concentration that occur during an inter dosing interval. Two types of nociceptive stimuli were used: (1) a cold pressor test (CP), (2) electrical stimulation (ES). Morphine was administered intravenously to achieve the two consecutive plasma concentrations. Blood samples were collected, concurrently with nociceptive responses, to determine plasma morphine concentrations. Methadone patients achieved mean C(SS1) and C(SS2) of 16 and 55 ng/ml respectively; those of controls were 11 and 33 ng/ml. Methadone patients were hyperalgesic to pain induced by CP but not ES. Despite significantly greater plasma morphine concentrations, methadone patients experienced minimal antinociception in comparison with controls. Furthermore in methadone patients, the antinociception ceased when the infusion ended. In comparison, the duration of effect in control subjects was 3 h. The fluctuations that occurred in plasma R(-)-methadone concentration during an inter-dosing interval had little effect on patients' responses to morphine. Our findings suggest that methadone patients are cross-tolerant to the antinociceptive effects of morphine, and conventional doses of morphine are likely to be ineffective in managing episodes of acute pain amongst this patient group. Further research is needed to determine whether other drugs are more effective than morphine in managing acute pain in this patient population. PMID- 11427328 TI - Despite clinical similarities there are significant differences between acute limb trauma and complex regional pain syndrome I (CRPS I). AB - In order to analyze the pathophysiology behind the clinical similarity acutely after limb trauma and in acute stages of complex regional pain syndrome (CRPS), 20 patients with external fixation after distal radius fracture (3.5 days after surgery) without signs of CRPS and 24 patients suffering from acute CRPS I (without nerve lesion; duration, 5 weeks) were investigated. Hyperalgesia to heat was tested by a feedback-controlled thermode, and to mechanical stimuli by an impact stimulator. The sympathetic nervous system was examined by measuring skin temperature (infra-red thermography), testing different sympathetic vasoconstrictor reflexes (laser-Doppler flowmetry) and quantitative sudometry after thermal load (thermoregulatory sweat test). We found hyperalgesia to heat after trauma (P<0.001), but not in CRPS, whereas mechanical hyperalgesia was present in both patient groups (trauma: P<0.001; CRPS: P<0.005). Skin temperature was significantly increased on the affected side in both patient groups (acute trauma: P<0.001; CRPS: P<0.005). However, sympathetic failure, as indicated by impairment of sympathetic vasoconstrictor reflexes (P<0.02) and hyperhidrosis (P<0.01), was found exclusively in CRPS patients. Our results indicate that pain and vasomotor disturbances may be generated by different mechanisms acutely after trauma and in acute CRPS. Despite the clinical similarity, additional changes in the peripheral or central nervous system are required for CRPS. In the light of our observations, it seems unlikely that CRPS is a simple exaggeration of post traumatic inflammation. PMID- 11427329 TI - The Faces Pain Scale-Revised: toward a common metric in pediatric pain measurement. AB - The Faces Pain Scale (FPS; Bieri et al., Pain 41 (1990) 139) is a self-report measure used to assess the intensity of children's pain. Three studies were carried out to revise the original scale and validate the adapted version. In the first phase, the FPS was revised from its original seven faces to six, while maintaining its desirable psychometric properties, in order to make it compatible in scoring with other self-rating and observational scales which use a common metric (0-5 or 0-10). Using a computer-animated version of the FPS developed by Champion and colleagues (Sydney Animated Facial Expressions Scale), psychophysical methods were applied to identify four faces representing equal intervals between the scale values representing least pain and most pain. In the second phase, children used the new six-face Faces Pain Scale-Revised (FPS-R) to rate the intensity of pain from ear piercing. Its validity is supported by a strong positive correlation (r=0.93, N=76) with a visual analogue scale (VAS) measure in children aged 5-12 years. In the third phase, a clinical sample of pediatric inpatients aged 4-12 years used the FPS-R and a VAS or the colored analogue scale (CAS) to rate pain during hospitalization for surgical and non surgical painful conditions. The validity of the FPS-R was further supported by strong positive correlations with the VAS (r=0.92, N=45) and the CAS (r=0.84, N=45) in this clinical sample. Most children in all age groups including the youngest were able to use the FPS-R in a manner that was consistent with the other measures. There were no significant differences between the means on the FPS-R and either of the analogue scales. The FPS-R is shown to be appropriate for use in assessment of the intensity of children's acute pain from age 4 or 5 onward. It has the advantage of being suitable for use with the most widely used metric for scoring (0-10), and conforms closely to a linear interval scale. PMID- 11427330 TI - A comparison of visual analog scale and categorical ratings of headache pain in a randomized controlled clinical trial with migraine patients. AB - A visual analog scale (VAS) method of assessing headache pain was compared with a standard categorical four-grade scale (4GS) in a randomized, placebo-controlled, double-blind, clinical trial involving 792 treated migraine outpatients who received oral rizatriptan 5 mg, sumatriptan 50 mg, or placebo for a moderate or severe headache. The VAS and 4GS were equally useful in demonstrating that the active drugs were superior to placebo at reducing headache pain, and in showing that the active drugs were similarly effective. For both rizatriptan and sumatriptan, slightly larger effect sizes were observed with the 4GS compared with the VAS. In analyses using data combined across all treatment groups, VAS and 4GS scores were highly correlated. Use of the VAS imposed additional administrative burdens. These findings suggest that the 4GS may be the preferred scale for assessing headache pain in clinical trials involving adult migraineurs. PMID- 11427331 TI - Gabapentin inhibits the substance P-facilitated K(+)-evoked release of [(3)H]glutamate from rat caudial trigeminal nucleus slices. AB - The effect of gabapentin on the release of the spinal sensory neurotransmitter glutamate has been investigated in an in vitro model using a perfused thin slice preparation from the rat brainstem containing the spinal trigeminal caudal subnucleus (Sp5C) and pre-incubated with [(3)H]glutamate. Addition of excess K(+) to the perfusing solution increased the content of tritium in the perfusate. The prior addition of substance P increased this index of glutamate release in a concentration-dependent manner, with the mean maximum of around 50% increase obtained at 1-3 microM. The action of substance P to increase the evoked release of glutamate was blocked by the antagonist CP-99994, suggesting a specific involvement of the NK(1) receptor in mediating the facilitatory effect. On its own, gabapentin at up to 100 microM did not modify the baseline level of K(+) evoked release of glutamate; however, gabapentin caused a concentration-dependent decrease of the facilitatory effect of substance P (EC(50)=6.49 microM). The R-( )- and S-(+)-isomers of 3-isobutylgaba were then tested against the increase in K(+)-evoked release of glutamate by substance P. S-(+)-3-isobutylgaba (pregabalin) at 30 microM acted like gabapentin to reduce the substance P mediated increase of release almost to the baseline level of K(+)-evoked release, while in contrast the R-(-)-isomer at this concentration produced no reduction, and rather a trend towards a further enhancement of the potentiating effect of substance P. In conclusion, we have found and characterized an effect of gabapentin that is of possible mechanistic relevance to the anti hyperalgesic/allodynic actions of this compound. PMID- 11427332 TI - Integrated brain activity in medial temporal and prefrontal areas predicts subsequent memory performance: human declarative memory formation at the system level. AB - After an era in which lesion studies have identified the declarative memory system and its essential anatomical structures, functional imaging and event related potential studies have begun to delineate the neural underpinnings of declarative memory formation at the system level. By memory formation, we refer to those mnemonic processes present during encoding that transform perceptual representations into enduring memories. Recent studies have revealed that distinct regions in medial temporal and prefrontal areas exhibit more neural activity during successful than unsuccessful memory formation. We attempt to identify the nature of the processes underlying these subsequent memory effects. Reviewed data suggest specific mnemonic operations in the medial temporal lobe that may be integrated with semantic/perceptual operations and subserving operations in the prefrontal cortex. The formation of relational and non relational memories may be supported by distinct subregions within these two brain regions. While the medial temporal lobe may have a serial organizational structure, with a processing hierarchy, interactions between medial temporal and prefrontal areas seem to occur in a parallel and bi-directional fashion. Interacting with this system, emotionally arousing events enhance neural activity in the amygdala, which in turn may modulate processing in other brain regions responsible for declarative memory formation. PMID- 11427333 TI - An approach to the complexity of the brain. AB - The establishment of ordered neuronal connections is supposed to take place under the control of specific cell adhesion molecules (CAM) which guide neuroblasts and axons to their appropriate destination. The extreme complexity of the nervous system does not provide a favorable medium for the development of deterministic connections. Simon's [112] theorems offer a mean to approach the high level of complexity of the nervous system. The basic tenet is that complex systems are hierarchically organized and decomposable. Such systems can arise by selective trial and error mechanisms. Subsystems in complex systems only interact in an aggregate manner, and no significant information is lost if the detail of aggregate interactions is ignored. A number of nervous activities, which qualify for these requirements, are shown. The following sources of selection are considered: internal and external feedbacks, previous experience, plasticity in simple structures, and the characteristic geometry of dendrites. The role played by CAMs and other membrane-associated molecules is discussed in the sense that they are either inductor molecules that turn on different homeobox genes, or downstream products of genes, or both. These molecules control cellular and tissular differentiation in the developing brain creating sources of selection required for the trial and error process in the organization of the nervous tissue. PMID- 11427334 TI - Effect of adrenomedullin receptor and calcitonin gene-related peptide receptor antagonists on centrally mediated adrenomedullin renal action. AB - To determine pharmacological specificity of intracerebroventricular (IVT) administration of adrenomedullin (AM) on water and sodium excretion, studies were performed in rats pretreated with AM (22-52), a putative AM-receptor antagonist or CGRP(8-37), a ligand that preferentially antagonizes the CGRP(1)-receptor subtype. In addition, the effects of IVT injection of calcitonin-gene-related peptide (CGRP) on urinary water and electrolyte excretion was assessed. Intracerebroventricular administration of rat adrenomedullin to conscious hydrated rats resulted in a significant increase in urinary volume and sodium excretion during the 6-h period of urine collection and was most effective at 3 and 6 h. Although less effective than AM, central administration of CGRP induced diuresis and natriuresis. Pretreatment with AM (22-52) or CGRP(8-37) significantly suppressed the diuretic and natriuretic effect of IVT-AM. These data suggest that both CGRP(1) and AM receptors are involved in the centrally mediated diuretic and natriuretic action of the AM. Our results provide evidence supporting the hypothesis that endogenous AM plays a role in the central nervous control of fluid and electrolyte homeostasis. PMID- 11427335 TI - Nitric oxide may underlie learned fear in the elevated T-maze. AB - The present study evaluated the role of nitric oxide (NO) in learned and innate fear in rats submitted to the elevated T-maze (ETM). Learned and innate fear were evaluated through the inhibitory avoidance and escape behaviour from the open arms, respectively. Rats treated with the inhibitor of NO synthesis N(omega) nitro-L-Arginine methyl ester (L-NAME; 5, 10, and 50 mg. kg(-1)) were able to learn the inhibitory avoidance. However, L-NAME (50 mg. kg(-1)), but not its inert isomer N(omega)-nitro-D-arginine methyl ester (D-NAME, 50 mg. kg(-1)), impaired the inhibitory avoidance 2 with no change in the baseline values, thus suggesting an anxiolytic-like effect without locomotor impairment. All treatments with L-NAME were able to induce increased mean arterial pressure (MAP), measured indirectly through the animal's tail. The treatment with L-NAME (5 and 10 mg. kg( 1)) failed to induce anxiolysis but significantly increased the MAP of the animals, which indicates that hypertension per se, did not underlie anxiolysis induced by L-NAME. L-Arginine, the precursor molecule for NO synthesis, facilitated the inhibitory avoidance and counteracted the L-NAME (50 mg. kg(-1)) induced anxiolysis. Neither previous treatment was able to change the escape behaviour. The results indicate that NO may underlie learned, but not innate, fear in the ETM. PMID- 11427336 TI - Role of wake inducing brain stem area on rapid eye movement sleep regulation in freely moving cats. AB - Some of the characteristic symptoms associated with rapid eye movement (REM) sleep are opposite to, while some apparently resemble, those of wakefulness. Therefore, it was hypothesised that the neurons present in the wakefulness inducing area(s) in the brain are likely to communicate with the REM sleep related neurons. Brain stem neurons were classified based on their firing rates in relation to electrophysiological correlates associated with spontaneous sleep and wakefulness recorded from freely moving, normally behaving cats. Thereafter, the responses of those classified neurons to stimulation of brain stem reticular wakefulness inducing area were studied. Results from 63 neurons showed that the wake inducing area affected 62% of the neurons. Fifty-eight percent of the neurons which increased firing during wakefulness, including the REM-OFF neurons, were excited, while 70% of the neurons which decreased firing during wakefulness, including the REM-ON neurons, were inhibited. These observations support our hypothesis and, along with their physiological significance, are discussed. PMID- 11427337 TI - Effects of intracerebroventricularly administered chimeric peptide of metenkephalin and FMRFa--[D-Ala2]YFa-on antinociception and its modulation in mice. AB - An enzymatically stable analog of YGGFMKKKFMRFamide (YFa), a chimeric peptide of metenkephalin and FMRFa, was synthesised. The antinociceptive effects of intracerebroventricular injections of this analog-[D-Ala2)]YAGFMKKKFMRFamide ([D Ala2]YFa)-was then investigated using the mouse radiant-heat tail-flick test. [D Ala2]YFa produced modest to good antinociception at 1, 2, and 5 microg/mouse (0.64, 1.28, and 3.22 nmol, respectively). This antinociceptive effect was completely reversed by the opioid receptor antagonist naloxone (1.5 microg/mouse: 4.12 nmol, intracerebroventricular [i.c.v.]), administered 5 min prior. Pretreatment (5 min) with either neuropeptides FF (1 microg/mouse: 0.92 nmol, i.c.v.) or FMRFa (1 microg/mouse: 1.69 nmol, i.c.v.) significantly attenuated the antinociceptive effects induced by [D-Ala2]YFa (1 microg/mouse, i.c.v.). Intracerebroventricular administration of [D-Ala2]YFa at 1 microg/mouse dose with morphine (2 microg/mouse: 5.86 nmol, i.c.v.) produced an additive antinociceptive effect, suggesting that [D-Ala2]YFa may have a modulatory effect on opioid (morphine) analgesia. These results provide further support for a role of such amphiactive sequences in antinociception and its modulation. PMID- 11427338 TI - Morphine tolerance in mice is independent of polymorphisms in opioid receptor sequences. AB - Pharmacogenomics links individual drug response variation to genetic differences, such as single nucleotide polymorphisms (SNPs). In particular, pharmacogenomics will allow clinicians to use genetic diagnostics to predict the response of a patient to a drug. We investigated whether SNPs in opioid receptors correlated with the development of morphine tolerance in mouse strains that showed either high or low tolerance to morphine. Sequencing identified five silent SNPs in the delta opioid receptor that varied from the published sequence in some strains, but which were found in both high and low tolerance strains. The mu and kappa opioid receptor sequences had no SNPs. Taken together, these data definitively demonstrate that morphine tolerance development in mice is independent of opioid receptor sequence. PMID- 11427339 TI - Elevation of guinea pig brain preprodynorphin mRNA expression and hypothalamic pituitary-adrenal axis activity by "binge" pattern cocaine administration. AB - The endogenous opioid system and the hypothalamic-pituitary-adrenal (HPA) axis have been implicated in many of the neurobiological effects of cocaine. Previous studies in our laboratory showed that "binge" pattern cocaine administration increases preprodynorphin (ppDyn) mRNA levels in the caudate putamen and circulating levels of corticosterone in the rat. The present study extended these findings to guinea pigs, a species known to have a kappa opioid receptor profile similar to that of humans. Male guinea pigs were treated with: (a) "binge" pattern cocaine for 7 days (subchronic) (3 x 15 mg/kg/day, hourly, intraperitoneal); (b) "binge" pattern saline for 5 days followed by "binge" pattern cocaine for 2 days (subacute); or (c) "binge" pattern saline for 7 days. Thirty minutes after the final injection, levels of ppDyn mRNA were quantitated in the nucleus accumbens, caudate putamen, frontal cortex, amygdala, hippocampus, and hypothalamus using a solution hybridization RNase protection assay. Regional distribution of ppDyn mRNA levels in the guinea pig brain was similar to that found in rat, with highest levels in the nucleus accumbens and caudate putamen. In the caudate putamen, ppDyn mRNA was significantly increased following either 2 days (38% increase) or 7 days (32% increase) of "binge" pattern cocaine administration as compared to saline-treated controls. No significant changes in ppDyn mRNA levels were found in any other brain region. Both subacute and subchronic "binge" cocaine administration significantly elevated plasma levels of adrenocorticotropin hormone (ACTH) and cortisol. However, the ACTH and cortisol increases were significantly blunted following 7 days of "binge" cocaine administration as compared to 2 days of drug treatment, reflecting the development of HPA tolerance or adaptation to repeated cocaine administration. Thus, the ppDyn mRNA and HPA responses to cocaine in guinea pigs are similar to those observed in rats. PMID- 11427340 TI - Lesion size and amphetamine hyperlocomotion after neonatal ventral hippocampal lesions: more is less. AB - Neonatal hippocampal lesions in rats produce behavioral and neurochemical abnormalities post-puberty that are used in animal models for developmentally linked pathology in schizophrenia. In one model, adult rats exhibit enhanced sensitivity to the locomotor-activating effects of amphetamine, if they had sustained excitotoxic lesions of the ventral hippocampus on post-natal day 7. The hippocampal elements responsible for these lesion-induced developmental changes have not been fully characterized. The present study assessed the locomotor activating effects of amphetamine in adult rats that on day 7 had sustained either sham or ibotenic acid lesions of the ventral hippocampus alone ("standard lesions"), or the ventral hippocampus plus surrounding portions of entorhinal cortex and dorsal hippocampus ("large lesions"). "Standard lesions" produced the expected "supersensitive" locomotor response to amphetamine, while "large lesions" did not. No differences between these lesion groups were observed in baseline levels of locomotor activity or habituation. These data suggest that models of enhanced behavioral sensitivity to dopamine agonists after neonatal hippocampal lesions require functionality in the entorhinal cortex and/or dorsal hippocampus. It is possible that the behavioral abnormalities in the "neonatal hippocampal lesion model" reflect, at least in part, aberrant function within spared elements of the hippocampal complex. PMID- 11427341 TI - A comparative neuroethological study of limbic seizures induced by Parawixia bistriata venom and kainic acid injections in rats. AB - A number of neurotoxins derived from arthropod venoms are known to show highly selective effects on nervous tissue. These neurotoxins have been proved to be extremely useful tools to investigate either convulsive or anticonvulsive mechanisms in the nervous system. In the present work, intracerebroventricular injection of the crude venom from the spider Parawixia bistriata (Araneae, Araneidae) in rats induced convulsive limbic seizures (head and forelimb myoclonus, as well as rearing and falling). Neuroethological analysis showed that the limbic seizures induced by the venom were different from those induced by kainic acid. Intravenous injection of the same venom did not induce seizures, but the neuroethological analysis showed an intensification of grooming behavior similar to a displaced activity. In conclusion, our experiments point that crude venom of P. bistriata may contain convulsant neurotoxins probably acting in limbic system structures. The mechanism of action of these neurotoxins may be different from simple activation of glutamatergic kainate receptors, as evidenced by a comparative neuroethological analysis of seizures induced by either venom or kainic acid. PMID- 11427342 TI - Prostaglandins play an important role in diurnal changes of tuberoinfundibular dopaminergic neuronal activity and prolactin secretion in ovariectomized, estrogen-treated rats. AB - Intracerebroventricular injection of prostaglandins (PGs) has long been shown to stimulate the secretion of prolactin (PRL). The action of specific PG and the underlying mechanism, however, remain inconclusive. In this study, we evaluated the involvement of PGs in the diurnal changes of the tuberoinfundibular dopaminergic (TIDA) neurons and PRL secretion in ovariectomized (OVX) rats treated with estrogen (E2). Prior treatment of indomethacin (50 mg/kg, subcutaneous) for 24 h significantly prevented the afternoon decrease of TIDA neuronal activity and blocked the PRL surge. Intracerebroventricular injection of PGE(1) (5 microg/3 microl/rat) at 1000 h significantly lowered the activity of TIDA neurons, while similar treatment of PGE2 was without effect. In OVX + E(2) rats pretreated with indomethacin, PGE(1) given at 15, but not 30 or 180 min before decapitation at 1500 h significantly reversed the effects of indomethacin on TIDA neuronal activity, while the blocked PRL surge was not reversed. The PGs also exhibited a stimulatory effect on nigrostriatal dopamine (DA) neurons with PGE2 being the most effective agent. These results indicate that endogenous PGs play a significant role in the control of central DA neurons, especially the diurnal changes of TIDA neuronal activity and PRL secretion in OVX + E2 rats. PMID- 11427343 TI - Expression of Fos in the piriform cortex after acquisition of olfactory learning: an immunohistochemical study in the rat. AB - The piriform cortex (PCx), the main area of the primary olfactory cortex, is assumed to play a role in olfactory memory. Involvement of this paleocortex in mnesic processes was investigated by using Fos immunocytochemistry after acquisition of a two-odor discrimination task. Trained rats had to associate one odor of a pair with water reward while pseudo-trained rats were randomly rewarded. We further used non-trained rats and home cage control animals to determine the effect of manipulation and basal Fos level respectively. Except in control rats, Fos immunoreactivity was mainly distributed in brain areas involved in olfactory processing, learning and arousal. The trained, pseudo-trained, and non-trained rats showed a high Fos labeling in the entire PCx. However, quantitative analysis demonstrated a statistically higher Fos immunoreactivity in the anterior PCx in comparison with the posterior PCx for these rats. Furthermore, behavioral data allowed us to distinguish two groups of trained rats according to the number of days required to acquire the task. Rats with slow acquisition showed a higher Fos immunoreactivity in the whole PCx in comparison with the rats exhibiting a fast acquisition. Our findings support the assumption of a PCx rostro-caudal heterogeneity which could sustain differential information processing. PMID- 11427344 TI - Caffeine reverses the memory disruption induced by intra-nigral MPTP-injection in rats. AB - The present study was carried out to test the possible effects of caffeine in improving the memory deficits observed in 1-methyl-4-phenyl-1,2,3,6 tetrahydropyrindine (MPTP)-lesioned rats, an animal model of early stage Parkinson's disease. Caffeine at the doses of 0.1-0.3 mg/kg (intraperitoneal) reversed the impairing effect of the administration of MPTP (1 micromol/side) into the substantia nigra, compact part, of rats on the avoidance scores in the training and test sessions of a two-way active avoidance task. This effect was not due to a motor or sensory alteration because the caffeine-induced learning and memory improvement was independent of the locomotor stimulant effect of the drug and there were no differences in the reaction time of the animals to a footshock (unconditioned stimulus) or a sound cue (conditioned stimulus) after caffeine treatment. These results suggest that the reported dopamine/adenosine receptor interaction can be used to restore defective learning and memory processes in Parkinson's disease and indicate that caffeine and other adenosine receptor antagonists are drugs with the potential for treatment of the cognitive disabilities of Parkinson's disease. PMID- 11427345 TI - Continuous, bilateral Achilles' tendon vibration is not detrimental to human walk. AB - Sensory feedback from the moving limbs contributes to the regulation of animal and human locomotion. However, the question of the specific role of the various modalities is still open. Further, functional loss of leg afferent fibres due to peripheral neuropathy does not always lead to major alteration in the gait pattern. In order to gain further insight on proprioceptive control of human gait, we applied vibratory tendon stimulation, known to recruit spindle primary afferent fibres, to both triceps surae muscles during normal floor walk. This procedure would disturb organisation and execution of walking, especially if spindles fire continuously and subjects are blindfolded. Vibration induced significant, though minor, changes in duration and length of stance and swing phase, and on speed of walking and kinematics of lower limb segments. No effect was induced on angular displacement of the ankle joint or trunk and head kinematics. This paucity of effects was at variance with the perception of the subjects, who reported illusion of leg stiffness and gait imbalance. These findings would speak for a selective gating of Ia input during locomotion and emphasise the notion that the central nervous system can cope with an unusual continuous input along the Ia fibres from a key muscle like the soleus. PMID- 11427346 TI - Microinjection of arginine vasopressin into the central nucleus of amygdala suppressed nociceptive jaw opening reflex in freely moving rats. AB - This study was performed to examine the antinociceptive effect after microinjection of arginine vasopressin (AVP) into the central nucleus of amygdala. We recorded the jaw opening reflex in freely moving rats. After injection of 0.2 or 0.4 nM AVP into the central nucleus of amygdala, digastric electromyogram (dEMG) was suppressed to 55 +/- 5% or 88 +/- 3 of the control. Artificial cerebrospinal fluid had no effects on the basal dEMG activity. V(1) vasopressin receptor antagonist blocked the suppressive effect produced by microinjection of 0.4 nM AVP from 53 +/- 3 to 81 +/- 3% of the control. However, V(2) vasopressin receptor antagonist did not affect changes in dEMG. We observed dEMG activity after intracerebroventricular injection of naloxone, methysergide, or phentolamine. All drugs did not affect the basal dEMG activity at our dose. Naloxone blocked the suppressive effect of 0.4 nM AVP from 42 +/- 4 to 79 +/- 5% of the control. Methysergide also inhibited the suppression of dEMG from 44 +/- 3 to 83 +/- 6% of the control. However, phentolamine, an alpha-adrenergic receptor antagonist, did not affect the suppression of dEMG. These results indicate AVP in the central nucleus of amygdala has potent analgesic effects in the orofacial area. The antinociception of central AVP seems to be mediated by opioid and serotonergic pathways. PMID- 11427347 TI - Mechanisms of transport across cell membranes of complexes contained in antitumour drugs. AB - Various mechanism of antitumour drug transport across cell membranes has been described. Particular attention has been paid to a passive transport, active transport and multidrug resistance of complexes contained in antitumour drugs. A drug supply to the target site depends on the blood circulation within the tumour, on characteristic drug diffusion in the tissue, and also on binding protein. The physiologic transfer of hydrophilic compounds across the membrane is usually intermediated by means of a specific receptor or a carrier in that membrane, which facilitates the transport of compounds to and from the cell. Some drugs, e.g. doxorubicin and annamycin, can pass across the membrane by intermediacy of liposomes which exhibit a great activity in penetrating into tumour cells. The efficiency of antitumour drugs is limited by the appearance of resistance, i.e. by the lack of sensitivity of the cell to the administered drug. The presence in the membrane of specific proteins belonging to the ABC carriers group is postulated in a resistance theory; they would be responsible for 'pumping out' lipophilic drug molecules from the cell. Participation of high energy ATP molecule is required by P-glycoprotein (Pgp) and by MRP protein described in this paper for their action. The mechanisms that are responsible for the cell resistance to drugs have been presented by analysing the resistance to antimetabolites, particularly to folate and fluoropyrimidine analogues, to alkylating agents, e.g. cisplatinum, and to heterocyclic compounds being responsible for so-called multidrug resistance. PMID- 11427348 TI - Substituted amylose as a matrix for sustained-drug release: a biodegradation study. AB - Substituted amylose polymers are prepared by reacting amylose chains with a suitable substituent such as 1,2-epoxypropanol (glycidol). Substituted amylose polymers are introduced as novel excipients for controlled release of bioactive materials. Since substituted amylose polymers are amylose-based polymers, they are subject to biodegradation by alpha-amylase enzymes present in the gastro intestinal tract; thus, gamma spectroscopy is used to follow the release of the natural abundant rhenium (VII) oxide used as a drug model, and to test their resistance to alpha-amylase enzymatic degradation. Two substituted amylose solid dosage forms were prepared: (i) matrix system and (ii) dry-coated tablets. Matrix systems and dry-coated tablets maintained their structure, and controlled the release of [186Re] showing no significant degradation of tablets by alpha amylase. PMID- 11427349 TI - The in-vitro porcine adhesion model is not predictive of the esophageal transit of risedronate tablets in humans. AB - Mucosal damage due to esophageal adhesion of pharmaceuticals is a continued concern to both health care providers and drug manufacturers. As a result of this concern, dosage forms are now being designed to exhibit minimal esophageal adhesion. Previous researchers have used an in-vitro porcine esophageal model to determine the propensity for formulations to adhere to the esophagus as an alternative to human scintigraphy studies. This study used a porcine esophageal adhesion model similar to that used previously to determine the adhesiveness of placebo bisphosphonate formulations. Results are analogous to those obtained by previous researchers, with film-coated tablets showing greater adhesiveness than uncoated tablets. These same tablet formulations were also evaluated previously by a human scintigraphy study, and the results were exactly opposite of those obtained using the in-vitro porcine model. In the human scintigraphy study, the film-coated placebo risedronate tablet had a faster transit time than an uncoated round placebo tablet. In conclusion, the in-vitro porcine esophageal model is not predictive of esophageal transit in man and gamma scintigraphy is the preferred method to evaluate esophageal transit. PMID- 11427350 TI - Effects of non-ionic surfactants as permeation enhancers towards piroxicam from the poloxamer gel through rat skins. AB - The enhancing effects of non-ionic surfactants on the permeation of piroxicam from the poloxamer gels were evaluated using Franz diffusion cells fitted with excised rat skins. The effectiveness of penetration enhancers, the ratio of piroxicam flux in the presence or absence of enhancers, was defined as the enhancement factor. Among the various non-ionic surfactants tested, polyoxyethylene-2-oleyl ether showed the highest enhancing effects with an enhancement factor of 2.84. To elucidate the mechanisms of the action of enhancers, thermal analysis and histological examinations were carried out. Thermal analysis reveals that various surfactants have different fluidizing effects on stratum corneum. Skin pretreated with the poloxamer 407 gels containing various surfactants showed a loosely layered stratum corneum and wide intercellular space. PMID- 11427351 TI - Influence of radiation sterilization on the stability of trifluorothymidine. AB - The influence of radiation sterilization on the stability of trifluorothymidine (TFT) was investigated. TFT was irradiated under ambient atmosphere with a 60Co source and with an electron accelerator at 25, 50, and 100 kGy, respectively. The radiation-induced effects were determined by chromatographic and spectroscopic methods as well as potentiometrically with a fluoride selective electrode. TFT was moderately stable to ionizing radiation. The degradation induced by electron beam irradiation was significantly (P=95%) smaller than by gamma-irradiation. The radiolysis products amounted to about 0.25% after electron-beam irradiation at 25 kGy, and to about 0.50% after gamma-irradiation, respectively. The main irradiation product was 5-trifluoromethyluracil (TFMU). In addition five further impurities were detected with HPLC. Identification of degradation products was performed using HPLC-ESI-MS. A degradation path of TFT after radiation sterilization was shown. PMID- 11427352 TI - The effect of alpha-tocopherol on the in vitro solubilisation of lipophilic drugs. AB - alpha-Tocopherol is an excellent solvent for many poorly soluble drugs. The aim of this work was to study whether or not the presence of alpha-tocopherol has an influence on the solubilisation of poorly soluble drugs in simulated intestinal fluids (SIF). The solubilizing capacity of mixed micelles containing alpha tocopherol towards three lipophilic drugs was investigated. The solubilisation of alpha-tocopherol in an aqueous micellar phase was increased by the addition of monoglycerides (MG) and free fatty acids (FFA), preferably of medium chain length, as compared to a simple bile salt solution. The addition of alpha tocopherol to mixed micellar solutions seems to have an effect on the solubilizing capacity, which can be correlated to the partition coefficient of the drug to be solubilised. A positive effect on the solubilisation of griseofulvin and felodipine was found. For a highly lipophilic drug (Lu28-179), a positive effect on solubilisation was observed only in media containing MG and FFA of medium chain length. Generally, alpha-tocopherol cannot be considered an important factor for the solubilisation of highly lipophilic drugs in SIF. The presence of lipolytic digestion products (LDP) of the proper chain length in relation to the drug to be solubilised is much more important. PMID- 11427353 TI - Evaluation of percutaneous absorption and skin irritation of ketoprofen through rat skin: in vitro and in vivo study. AB - The influences of different mechanisms of penetration enhancers (such as menthol, azone, ethanol and nonivarnide) regarding the percutaneous absorption and skin irritation of ketoprofen formulations through rat skin were investigated by in vitro and in vivo study. The skin irritation degree at the end of the experiment (10 h) was deterinined by pathologic biopsy and colorimetry methods. In vitro, the menthol showed the most potent enhancing effect. Furthermore, the enhancement effect of a combination of menthol and nonivamide was higher than that of their individual use alone. In vivo the formulation containing 0.05% nonivantide, 5% menthol and 20% ethanol showed a higher penetration rate and an acceptable degree of skin irritation compared to a commercial product (Formax plus gel containing 3% ketoprofen), indicating that it could be used in the clinical situation. PMID- 11427354 TI - Two glycine containing 2-chloroethylnitrosoureas--a comparative study on some physicochemical properties, in vivo antimelanomic effects and immunomodulatory properties. AB - Physicochemical properties such as alkylating and carbamoylating activity and in vivo antimelanomic effects against B16 melanoma of the spin labeled (nitroxyl free radical containing) glycine nitrosourea (SLCNUgly) and its nonlabeled analogue (ChCNUgly), synthesized in our laboratory are studied and compared to those of antitumour drug 3-cyclohexyl-1-(2-chloroethyl)-1-nitrosourea (CCNU). We have demonstrated that introducing of glycine moiety in the nitrosourea structure in practice does not affect either alkylating or carbamoylating activity. On the other hand replacement of cyclohexyl moiety in ChCNUgly structure with nitroxyl free radical leads to a decrease in carbamoylating activity and an increase in alkylating activity. Compound ChCNUgly showed in vivo a higher antimelanomic activity against B16 melanoma in comparison with CCNU and SLCNUgly. It completely inhibited B16 melanoma growth (TGI=100%) at a dose 64.0 mg/kg. Moreover, we established that joint i.p. application in normal mice of SLCNUgly plus a new immunostimulator (C3bgp) formerly isolated in our laboratory led to a 75% restoration in immune function with respect to antibody production measured by Jerne hemolytic plaque assay. In contrast, no immunostimulation was found after joint application of C3bgp plus ChCNUgly or CCNU at the same experimental conditions. Based on these preliminary results, a possibility for developing of new combination immunochemotherapy schemes for treatment of human cancers is discussed. PMID- 11427355 TI - Magnetic resonance imaging investigation of the mixing-segregation process in a pharmaceutical blender. AB - Magnetic Resonance Imaging (MRI) was used to study the mixing process of binary mixtures of free flowing sugar beads in a Turbula mixer. In order to make particles MRI-sensitive, some reference beads were doped with an organic oil. Doped and undoped particles were mixed and MRI was used to non-destructively image the particle bed for a given number of mixer rotations (NR), bead diameter ratio (R=d(ref)/d(i)) and rotation speed (V). All the results were quantified on the basis of image analysis to characterise the degree of mixing. Studies showed that for binary mixtures of identical particle size, the mixing was complete after 30 rotations, whereas for beads of different size (R=2.8) a segregated steady state was obtained after nearly 10 rotations. Experiments revealed that segregation appeared as soon as R=0.9. Moreover, the lower the rotation speed, the more segregated the final state was. It appeared that for a filling level greater than 80%, dead regions appeared in the centre of the powder bed. In conclusion, when the particles are non-cohesive, the Turbula blender perfectly mixes identical beads but segregation occurs for beads of different size after just a few rotations. PMID- 11427356 TI - Effect of some physiological and non-physiological compounds on the phase transition temperature of thermoresponsive polymers intended for oral controlled drug delivery. AB - Poly-N-isopropylacrylamide (PNIPAAm) thermosensibility makes this polymer a very attractive candidate for controlled drug delivery systems. The polymer possesses a lower critical solution temperature (LCST) which was found to be around 32 degrees C in pure water, but which can be affected by the medium composition, i.e. presence of salts or surfactants. The knowledge of the effects of such substances on the LCST is very important while using PNIPAAm as a controlled drug delivery agent. The influence of a number of physiological and non-physiological salts and surfactants has been studied. The results obtained show that the addition of salts provokes an important decrease of the LCST of the polymer (salting out effect). A strong influence of the valence and of the size of the anions of the halide group was found. As to the surfactants, according to their type and concentration, a decrease or an increase of the LCST or even no effect at all were found. The effect of the GI secretions on the PNIPAAm phase separation temperature is also discussed. PMID- 11427357 TI - Stability of ascorbyl palmitate in topical microemulsions. AB - Ascorbyl palmitate and sodium ascorbyl phosphate are derivatives of ascorbic acid, which differ in stability and hydro-lipophilic properties. They are widely used in cosmetic and pharmaceutical preparations. In the present work the stability of both derivatives was studied in microemulsions for topical use as carrier systems. The microemulsions were of both o/w and w/o types and composed of the same ingredients. The stability of the less stable derivative ascorbyl palmitate was tested under different conditions to evaluate the influence of initial concentration, location in microemulsion, dissolved oxygen and storage conditions. High concentrations of ascorbyl palmitate reduced the extent of its degradation. The location of ascorbyl palmitate in the microemulsion and oxygen dissolved in the system together significantly influence the stability of the compound. Light accelerated the degradation of ascorbyl palmitate. In contrast, sodium ascorbyl phosphate was stable in both types of microemulsions. Sodium ascorbyl phosphate is shown to be convenient as an active ingredient in topical preparations. In the case of ascorbyl palmitate, long-term stability in selected microemulsions was not adequate. To formulate an optimal carrier system for this ingredient other factors influencing the stability have to be considered. PMID- 11427358 TI - Comparison of the physicochemical properties of the N-(2-hydroxyethyl) pyrrolidine, diethylamine and sodium salt forms of diclofenac. AB - Non steroidal anti-inflammatory agents (NSAIDs) such as diclofenac have very low aqueous solubilities and consequently salt formation may be used to enhance solubility and dissolution rate. In this study, we examined the physicochemical properties of three diclofenac salts, diclofenac sodium (DNa), diclofenac N-(2 hydroxyethyl)pyrrolidine (DHEP) and diclofenac diethylamine (DDEA), and their different solid state forms to determine the influence of salt form on solubility, dissolution rate and membrane transport. The equilibrium solubility of DDEA at 25 degrees C was determined as 33 mM, lower than the solubilities of DHEP (273 mM) and DNa (66 mM) previously reported (Ledwidge and Corrigan, 1998). In addition to the dihydrate form of DHEP previously characterised, monohydrate forms of DHEP and DDEA were identified. Intrinsic dissolution rate studies were used to determine the solubility ratios of the hydrated and anhydrous forms. The monohydrate form of DHEP was found to be 1.8 times less soluble than the anhydrate, whereas DDEA anhydrate was approximately 1.7 times as soluble as the monohydrate form. On investigation of the pH-solubility profile (25 degrees C) of DDEA, appreciable supersaturation (76 mM) relative to the theoretical profile, was detected at the pH(max). This contrasts with values of >800 and 67 mM for DHEP and DNa, respectively. The transport of salt solutions through a porous membrane (Visking) was investigated. A linear relationship between concentration (mM) and rate of transport (mmol/h) was established for DNa and DHEP solutions. The mass transfer coefficient determined for DHEP was lower than that for the other two salts. Nevertheless, the maximum transport rate obtained for DHEP is almost six times higher than that obtained for DDEA. PMID- 11427359 TI - The use of scintigraphy to demonstrate the rapid esophageal transit of the oval film-coated placebo risedronate tablet compared to a round uncoated placebo tablet when administered with minimal volumes of water. AB - As our population ages, and the consumption of pharmaceutical products rises, the incidence of solid oral dosage forms lodging in the esophagus is likely to increase and may be formulation dependent. The aim of this study was to compare the esophageal transit of the commercial film-coated risedronate tablet and a round uncoated tablet resembling the alendronate 10 mg tablet which is reported to cause esophagitis if ingested with little to no water. Water volumes of 30 ml and 50 ml were selected as these volumes can detect formulations prone to esophageal adhesion and a habits and practice study showed that these volumes are within the range preferred by women (7-385 ml). A total of 28 healthy postmenopausal women completed the four-way crossover scintigraphy study. For both volumes of water, the film-coated placebo risedronate tablet had a statistically significant faster esophageal transit time than the uncoated placebo tablet (P=0.002 for 30 ml water and P<0.001 for 50 ml water). Among those taking the round, flat, uncoated tablet, five subjects had esophageal stasis (transit >20 s) and in three subjects the tablet remained in the esophagus at the end of the 10-min imaging period. No stasis was observed for the oval film-coated placebo risedronate tablet. This study demonstrates that tablet size, shape and coating are pharmaceutical parameters which can be controlled to minimize esophageal contact of a dosage form with esophageal tissue. PMID- 11427360 TI - Pharmacokinetics of acetaminophen in Hong Kong Chinese subjects. AB - The pharmacokinetics of acetaminophen have been well studied in different populations, especially in Caucasians. However, limited studies on acetaminophen pharmacokinetics have been conducted in the native Chinese and few such data have been reported in the English language literature. Previous published studies suggested that environmental and genetic factors may cause inter-individual difference in acetaminophen disposition, thus we investigated the pharmacokinetics of acetaminophen in Hong Kong Chinese subjects. A single 500 mg oral dose of acetaminophen was administered to 12 healthy male Chinese subjects under fasting conditions. Multiple blood samples were obtained after drug administration. Plasma acetaminophen concentrations were determined using HPLC, and its main pharmacokinetic parameters were generated. In comparison to other published data, acetaminophen half-life was considerably longer (15-62%), and oral clearance was lower (16-56%) in Hong Kong Chinese as compared to Australian Chinese, Caucasians (USA, UK, Australia), and subjects from Pakistan, Denmark, Spain and South Africa. Similarities however were found in the pharmacokinetic parameters between Hong Kong Chinese and Mainland Chinese subjects. The observed pharmacokinetic parameters of acetaminophen in Hong Kong Chinese subjects may be different from other ethnic populations. Further studies are needed to verify this hypothesis. PMID- 11427361 TI - Kinematic comparison of Hybrid II test dummy to wheelchair user. AB - Hybrid test dummies provide a safe alternative to human subjects when investigating mechanisms of wheelchair tips and falls. The data that researchers acquire from these test dummies are more useful if the test dummy represents the population being studied. The goal of this study was to measure the validity of a 50th percentile Hybrid II test dummy (HTD) as an accurate representation of a wheelchair user. A test pilot with T8 paraplegia due to traumatic spinal cord injury served as a basis for validation. Simple modifications were made to the HTD to approximate the trunk stability characteristics of a person with a spinal cord injury. An HTD, a modified HTD, and a human test pilot were seated in an electric-powered wheelchair and several braking tests performed. The standard HTD underestimated the kinematics when compared to the test pilot. The modified HTD had less trunk stability than the standard HTD during all braking methods. The modified HTD and wheelchair test pilot had similar trunk stability characteristics during kill switch and joystick full-reverse braking conditions. The modified HTD is a satisfactory representation of a wheelchair user with a spinal cord injury; however, the modified test dummy underestimates the trunk dynamics during the less extreme joystick release braking. Work should continue on the development of a low-speed, low-impact test dummy that emulates the wheelchair user population. PMID- 11427362 TI - Evaluation of wheelchair drop seat crashworthiness. AB - Wheelchair seating crash performance is critical to protecting wheelchair users who remain seated in their wheelchairs during transportation. Relying upon computer simulation and sled testing seat loads associated with a 20 g/48 kph (20 g/30 mph) frontal impact and 50th percentile male occupant were estimated to develop test criteria. Using a static test setup we evaluated the performance of various types of commercially available drop seats against the loading test criteria. Five different types of drop seats (two specimens each) constructed of various materials (i.e. plastics, plywood, metal) were evaluated. Two types of drop seats (three of the total 10 specimens) met the 16650 N (3750 lb) frontal impact test criteria. While additional validation of the test protocol is necessary, this study suggests that some drop seat designs may be incapable of withstanding crash level loads. PMID- 11427363 TI - A kinetic analysis of trained wheelchair racers during two speeds of propulsion. AB - The purpose of the study was to investigate the propulsion kinetics of wheelchair racers at racing speeds and to assess how these change with an increase in speed. It was hypothesised that propulsive force would increase in proportion to speed, to accommodate the additional work required. Six wheelchair racers volunteered to participate in this study which required each athlete to push a racing wheelchair at 4.70 and 5.64 m s(-1) on a wheelchair ergometer (WERG). Eight pairs (16 in total) of strain gauges, mounted on four bars attached to the hand-rim of a racing wheelchair wheel, measured the medio-lateral and tangential forces applied to the hand-rim. Kinetic data were sampled at 200 Hz while a single on-line (ELITE) infrared camera operating at 100 Hz was positioned perpendicular to the WERG to record the location of the hand with respect to the hand-rim. In general, peak tangential force occurred when the hand was positioned on the hand-rim between 140 and 180 degrees. With the increase in speed, the peak hand-rim forces applied tangentially increased from 132 to 158 N and those applied medio laterally increased from 90 to 104 N. The ratio of tangential to total measured force was similar at both speeds (80 and 82%, respectively). In conclusion, these data indicate that wheelchair racers adopt a different propulsion strategy than that employed in everyday chairs and that the forces increase in proportion to propulsion speed. PMID- 11427364 TI - Does computer game play aid in motivation of exercise and increase metabolic activity during wheelchair ergometry? AB - GAME(Wheels) is an interface between a portable roller system and a computer that enables a wheelchair user to play commercially available computer video games. The subject controls the game play with the propulsion of their wheelchair's wheels on the rollers. The purpose of this study was to investigate whether using the GAME(Wheels) System during wheelchair propulsion exercise can help increase the individual's physiological response and aid in the motivation to exercise. Fifteen subjects participated in this study. The subjects propelled their wheelchairs on a portable roller that was equipped with the GAME(Wheels) System. There were two exercise trials consisting of 2 min of warm-up, 16 min of exercise and 2 min of cool-down. Physiological data (ventilation rate, oxygen consumption, heart rate) were collected. A significant difference (P<0.05) was found between exercise with GAME(Wheels) versus without GAME(Wheels) for average ventilation rate and average oxygen consumption. The differences were found during time periods of transition from warm-up to exercise, and before and after the midpoint of exercise. Written questionnaires showed that 87% of the individuals tested reported the system would help them work out on a regular basis. Playing the video game helped these individuals to reach their exercise training zone faster and maintain it for the entire exercise trial. PMID- 11427365 TI - The effect of knee-flexion angle on wheelchair turning. AB - The increasingly popular hyperflexed knee-flexion angle was evaluated to determine its effects on wheelchair turning. Twenty able-bodied subjects were tested comparing the effect of full knee extension and full knee flexion on a number of parameters. We empirically measured the angular velocity of subjects spinning 720 degrees in place, subjects' perceived ease of wheelchair turning, the overall length of the wheelchair, the anteroposterior position of the center of mass (COM), rolling resistance, turning resistance and rear-wheel traction. The combined moment of inertia of the wheelchair and system was modeled. We found that, in comparison with full extension, fully flexing the knees increased angular velocity by 40% and was perceived to be 66% easier by subjects. Overall length decreased by 39%, COM moved rearward 38%, rolling and turning resistance decreased by 21% and 17% respectively, rear-wheel traction increased by 12% and moment of inertia decreased by 42%. All empirically tested parameters were statistically significant (p<0.007). We conclude that the knee-flexion angle has a significant effect on wheelchair turning. The implications of these findings for wheelchair design and prescription will need to be validated on actual wheelchair users and for smaller increments in knee-flexion range. PMID- 11427366 TI - An autoregressive modeling approach to analyzing wheelchair propulsion forces. AB - Biomechanical signals collected during wheelchair propulsion are often analyzed by computing averages and/or peak values over several strokes. Due to the complex nature of the signals, this type of analysis may not be specific to identifying factors that may predispose a wheelchair user to joint pain/injury. Hence, a new technique is introduced that uses a system identification approach, autoregressive (AR) modeling, to analyze wheelchair propulsion force waveforms. In this application an AR method was used to create a model force waveform based on current and past values of digital pushrim force data. The feasibility of the AR modeling method over point-wise methods to detect asymmetry among force waveforms was tested with a group of 20 wheelchair users. Subjects propelled at a constant 0.9 m/s on a roller system during which 20 s of force data were collected from the SMART(Wheel)s, force and torque sensing pushrims. Both methods showed that the wheelchair users as a group propelled evenly, however, individual analysis using the AR model error estimates indicated that twenty-five percent demonstrated significant asymmetry in their force waveforms. If only point-wise means and variances of the applied bilateral forces were considered, most subjects would have appeared symmetrical. Thus, the AR modeling approach is more sensitive to detecting anomalies in propulsion technique. PMID- 11427368 TI - Is Alzheimer's disease a form of cancer? PMID- 11427369 TI - Serious infections stunt tumour growth. PMID- 11427370 TI - Unravelling metabolic syndrome X. PMID- 11427373 TI - Crispin Kirkman discusses the European biotech industry. Interviewed by Ben Ramster and Rebecca N. Lawrence. PMID- 11427374 TI - Are microwave instruments just expensive hot-plate stirrers? AB - The Discussion Forum provides a medium for airing your views on any issues related to the pharmaceutical industry and obtaining feedback and discussion on these views from others in the field. You can discuss issues that get you hot under the collar, practical problems at the bench, recently published literature, or just something bizarre or humorous that you wish to share. Publication of letters in this section is subject to editorial discretion and company promotional letters will be rejected immediately. Furthermore, the views provided are those of the authors and are not intended to represent the views of the companies they work for. Moreover, these views do not reflect those of Elsevier, Drug Discovery Today or its editorial team. Please submit all letters to Dr Rebecca Lawrence, News & Features Editor, Drug Discovery Today, e-mail: Rebecca.Lawrence@drugdiscoverytoday.com PMID- 11427375 TI - Drug discoverers - you need us! - Reply. PMID- 11427376 TI - Too many targets, not enough target validation - Reply. PMID- 11427377 TI - Is caspase-3 inhibition a valid therapeutic strategy in cerebral ischemia? AB - Neurodegenerative diseases are characterized by progressive impairment of brain function as a consequence of ongoing neuronal cell death. Apoptotic mechanisms have been implicated in this process and a major involvement of caspase-3, a typical pro-apoptotic executioner protease, has been claimed. In this review, the role of caspase-3 in neuronal cell loss in animal models of stroke is discussed and critically evaluated. In summary, it is concluded that the biochemical evidence favoring caspase-3 as a therapeutic target in cerebral ischemia is not convincing, and the development of selective caspase-3 inhibitors for the treatment of human stroke must be viewed as high risk. PMID- 11427378 TI - A genomic perspective on human proteases as drug targets. AB - Of the approximately 400 known human proteases, approximately 14% are under investigation as drug targets. Although the total is certain to rise during the finishing phase of the human genome project, the initial annotation of the approximately 30,000 human proteome set includes approximately 500 proteases. Bioinformatic analysis can now be performed on complete human protease families and will soon include comparisons with mice and fish. New sequences will require evaluation of their function in normal physiology and human disease. By revealing details such as splice variants and population polymorphisms, genomic sequence information will have a central role in the validation of protease drug targets. PMID- 11427379 TI - Clinical efforts to modulate angiogenesis in the adult: gene therapy versus conventional approaches. AB - A gene therapy approach towards the modulation of neovascularization provides important advantages that could be crucial for the success of therapies that target blood vessels. These advantages include sustained local expression and the ability to supply multiple pro- or anti-angiogenic factors. There is potential near-term success in the application of this approach for the treatment of ischemic vascular diseases. Although there is convincing proof of concept in animal models that an anti-angiogenesis gene therapy approach can be used to treat cancer, this is a highly competitive field with small molecules, recombinant proteins and monoclonal antibodies already in clinical trials. The scientific rationale for the use of gene therapy is sound, but realization of its full potential for the treatment of a broad array of diseases will require several challenging technical hurdles to be overcome and safety concerns to be alleviated. PMID- 11427380 TI - Monitor: molecules and profiles. AB - Monitor provides an insight into the latest developments in drug discovery through brief synopses of recent presentations and publications together with expert commentaries on the latest technologies. There are two sections: Molecules summarizes the chemistry and the pharmacological significance and biological relevance of new molecules reported in the literature and on the conference scene; Profiles offers commentary on promising lines of research, emerging molecular targets, novel technology, advances in synthetic and separation techniques and legislative issues. PMID- 11427382 TI - Combinatorial chemistry. PMID- 11427381 TI - Novel antitumour molecules. PMID- 11427383 TI - Air pollution and child mortality: a time-series study in Sao Paulo, Brazil. AB - Although most available evidence relating air pollution and mortality was obtained for adults, pollution has been also associated with increased mortality in children, but in a significantly smaller number of studies. This study was designed to evaluate the association between child mortality and air pollution in the city of Sao Paulo, Brazil, from 1994 to 1997. Daily records of mortality due to respiratory diseases for children under 5 years of age were obtained from the municipal mortality information improvement program. Daily concentrations of sulfur dioxide (SO(2)), carbon monoxide (CO), inhalable particulate matter less than 10 microm in diameter (PM(10)), and ozone were obtained from the state air pollution controlling agency. Information on minimum daily temperature and on relative humidity were obtained from the Institute of Astronomy and Geophysics of the University of Sao Paulo. Statistical analysis was performed through generalized additive models considering a Poisson response distribution and a log link. Explanatory variables were time, temperature, humidity, and pollutant concentrations. The loess smoother was applied to time (in order to model seasonality) and temperature. Significant associations between mortality and concentrations of CO, SO(2), and PM(10) were detected. The coefficients (and standard errors) of these three pollutants were 0.0306 (0.0076), 0.0055 (0.0016), and 0.0014 (0.0006), respectively. The observed associations were dose dependent and quite evident after a short period of exposure (2 days). According to the proposed model and considering the mean of the pollutant concentration during the period of the study, the estimated proportions of respiratory deaths attributed to CO, SO(2), and PM(10), when considered individually, are around 15, 13, and 7%, respectively. PMID- 11427384 TI - Relation between ambient air pollution and low birth weight in the Northeastern United States. AB - We evaluated the relation between term low birth weight (LBW) and ambient air levels of carbon monoxide (CO), particulate matter up to 10 microm in diameter (PM(10)), and sulfur dioxide (SO(2)). The study population consisted of singleton, term live births (37-44 weeks of gestation) born between 1 January 1994 and 31 December 1996 in six northeastern cities of the United States: Boston, Massachusetts; Hartford, Connecticut; Philadelphia, Pennsylvania; Pittsburgh, Pennsylvania; Springfield, Massachusetts; and Washington, DC. Birth data were obtained from National Center for Health Statistics Natality Data Sets. Infants with a birth weight < 2,500 g were classified as LBW. Air monitoring data obtained from the U.S. Environmental Protection Agency were used to estimate average trimester exposures to ambient CO, PM(10), and SO(2). Our results suggest that exposures to ambient CO and SO(2) increase the risk for term LBW. This risk increased by a unit increase in CO third trimester average concentration [adjusted odds ratio (AOR) 1.31; 95% confidence interval (CI) 1.06,1.62]. Infants with SO(2) second trimester exposures falling within the 25 and < 50th (AOR 1.21; CI 1.07,1.37), the 50 to < 75th (AOR 1.20; CI 1.08,1.35), and the 75 to < 95th (AOR 1.21; CI 1.03,1.43) percentiles were also at increased risk for term LBW when compared to those in the reference category (< 25th percentile). There was no indication of a positive association between prenatal exposures to PM(10) and term LBW. Increased ambient levels of air pollution may be associated with an increased risk for LBW. PMID- 11427385 TI - Urban air pollution and health inequities: a workshop report. AB - Over the past three decades, an array of legislation with attendant regulations has been implemented to enhance the quality of the environment and thereby improve the public's health. Despite the many beneficial changes that have followed, there remains a disproportionately higher prevalence of harmful environmental exposures, particularly air pollution, for certain populations. These populations most often reside in urban settings, have low socioeconomic status, and include a large proportion of ethnic minorities. The disparities between racial/ethnic minority and/or low-income populations in cities and the general population in terms of environmental exposures and related health risks have prompted the "environmental justice" or "environmental equity" movement, which strives to create cleaner environments for the most polluted communities. Achieving cleaner environments will require interventions based on scientific data specific to the populations at risk; however, research in this area has been relatively limited. To assess the current scientific information on urban air pollution and its health impacts and to help set the agenda for immediate intervention and future research, the American Lung Association organized an invited workshop on Urban Air Pollution and Health Inequities held 22-24 October 1999 in Washington, DC. This report builds on literature reviews and summarizes the discussions of working groups charged with addressing key areas relevant to air pollution and health effects in urban environments. An overview was provided of the state of the science for health impacts of air pollution and technologies available for air quality monitoring and exposure assessment. The working groups then prioritized research needs to address the knowledge gaps and developed recommendations for community interventions and public policy to begin to remedy the exposure and health inequities. PMID- 11427386 TI - The spatial association between community air pollution and mortality: a new method of analyzing correlated geographic cohort data. AB - We present a new statistical model for linking spatial variation in ambient air pollution to mortality. The model incorporates risk factors measured at the individual level, such as smoking, and at the spatial level, such as air pollution. We demonstrate that the spatial autocorrelation in community mortality rates, an indication of not fully characterizing potentially confounding risk factors to the air pollution-mortality association, can be accounted for through the inclusion of location in the model assessing the effects of air pollution on mortality. Our methods are illustrated with an analysis of the American Cancer Society cohort to determine whether all cause mortality is associated with concentrations of sulfate particles. The relative risk associated with a 4.2 microg/m(3) interquartile range of sulfate distribution for all causes of death was 1.051 (95% confidence interval 1.036-1.066) based on the Cox proportional hazards survival model, assuming subjects were statistically independent. Inclusion of community-based random effects yielded a relative risk of 1.055 (1.033, 1.077), which represented a doubling in the residual variance compared to that estimated by the Cox model. Residuals from the random-effects model displayed strong evidence of spatial autocorrelation (p = 0.0052). Further inclusion of a location surface reduced the sulfate relative risk and the evidence for autocorrelation as the complexity of the location surface increased, with a range in relative risks of 1.055-1.035. We conclude that these data display both extravariation and spatial autocorrelation, characteristics not captured by the Cox survival model. Failure to account for extravariation and spatial autocorrelation can lead to an understatement of the uncertainty of the air pollution association with mortality. PMID- 11427387 TI - Air pollution and respiratory symptoms: results from three panel studies in Bangkok, Thailand. AB - Several studies in North American cities have reported associations between air pollution and respiratory symptoms. Replicating these studies in cities with very different population and weather characteristics is a useful way of addressing uncertainties and strengthening inferences of causality. To this end we examined the responses of three different panels to particulate matter (PM) air pollution in Bangkok, Thailand, a tropical city characterized by a very warm and humid climate. Panels of schoolchildren, nurses, and adults were asked to report daily upper and lower respiratory symptoms for 3 months. Concentrations of daily PM(10) (PM with a mass median aerodynamic diameter less than 10 microm) and PM(2.5) (airborne particles with aerodynamic diameters less than 2.5 microm) were collected at two sites. Generally, associations were found between these pollution metrics and the daily occurrence of both upper and lower respiratory symptoms in each of the panels. For example, an interquartile increase of 45 microg/m(3) in PM(10) was associated with about a 50% increase in lower respiratory symptoms in the panel of highly exposed adults, about 30% in the children, and about 15% in the nurses. These estimates were not appreciably altered by changes in the specification of weather variables, stratification by temperature, or inclusion of individual characteristics in the models; however, time trends in the data cause some uncertainty about the magnitude of the effect of PM on respiratory symptoms. These pollutants were also associated with the first day of a symptom episode in both adult panels but not in children. The estimated odds ratios are generally consistent with and slightly higher than the findings of previous studies conducted in the United States. PMID- 11427388 TI - Reassessment of the lethal London fog of 1952: novel indicators of acute and chronic consequences of acute exposure to air pollution. AB - This article develops and assesses novel indicators of respiratory and other morbidity and mortality following London's lethal smog in the winter of 1952. Public health insurance claims, hospital admission rates for cardiac and respiratory disease, pneumonia cases, mortality records, influenza reports, temperature, and air pollutant concentrations are analyzed for December-February 1952-1953 and compared with those for the previous year or years. Mortality rates for the smog episode from December 1952 to February 1953 were 50-300% higher than the previous year. Claims that the smog only elevated health risks during and immediately following the peak fog 5-9 December 1952 and that an influenza epidemic accounted fully for persisting mortality increases in the first 2 months of 1953 are rejected. We estimate about 12,000 excess deaths occurred from December 1952 through February 1953 because of acute and persisting effects of the 1952 London smog. Pollution levels during the London smog were 5-19 times above current regulatory standards and guidelines and approximate current levels in some rapidly developing regions. Ambient pollution in many regions poses serious risks to public health. PMID- 11427389 TI - Acute pulmonary toxicity of particulate matter filter extracts in rats: coherence with epidemiologic studies in Utah Valley residents. AB - Epidemiologic reports by C.A. Pope III et. al. demonstrated that in the Utah Valley, closure of an open-hearth steel mill over the winter of 1987 was associated with reductions in respiratory disease and related hospital admissions in valley residents. To better examine the relationship between plant-associated changes in ambient particulate matter (PM) and respiratory health effects, we obtained total suspended particulate filters originally collected near the steel mill during the winter of 1986 (before closure), 1987 (during closure), and 1988 (after plant reopening). PM subcomponents were water-extracted from these filters and Sprague-Dawley rats were intratracheally instilled with equivalent masses of extract. Data indicated that 24 hr later, rats exposed to 1986 or 1988 extracts developed significant pulmonary injury and neutrophilic inflammation. Additionally, 50% of rats exposed to 1986 or 1988 extracts had increased airway responsiveness to acetylcholine, compared to 17 and 25% of rats exposed to saline or the 1987 extract, respectively. By 96 hr, these effects were largely resolved except for increases in lung lavage fluid neutrophils and lymphocytes in 1986 extract-exposed rats. Analogous effects were observed with lung histologic assessment. Extract analysis using inductively coupled plasma-mass spectroscopy demonstrated in all three extracts nearly 70% of the mass appeared to be sodium based salts derived from the glass filter matrix. Interestingly, relative to the 1987 extract, the 1986/1988 extracts contained more sulfate, cationic salts (i.e., calcium, potassium, magnesium), and certain metals (i.e., copper, zinc, iron, lead, strontium, arsenic, manganese, nickel). Although total metal content was (3/4) 1% of the extracts by mass, the greater quantity detected in the 1986 and 1988 extracts suggests metals may be important determinants of the pulmonary toxicity observed. In conclusion, the pulmonary effects induced by exposure of rats to water-based extracts of local ambient PM filters were in good accord with the cross-sectional epidemiologic reports of adverse respiratory health effects in Utah Valley residents. PMID- 11427390 TI - Air pollution and blood markers of cardiovascular risk. AB - Recent studies have linked air pollution to tens of thousands of premature cardiovascular deaths per year. The mechanisms of such associations remain unclear. In this study we examine the association between blood markers of cardiovascular risk and air pollution in a national sample of the U.S. population. Air pollution concentrations were merged to subjects in the Third National Health and Nutrition Examination Survey (NHANES III) in the United States, and the association with fibrinogen levels and counts of platelets and white blood cells were examined. The subjects in NHANES III are a representative sample of the U.S. population. Regressions controlled for age, race, sex, body mass index, current smoking, and number of cigarettes per day. The complex survey design was dealt with using mixed models with a random sampling site effect. In single-pollutant models, PM(10) (particulate matter with a mass median aerodynamic diameter less than 10 microm) was associated with all three outcomes (p< 0.05): Sulfur dioxide (SO(2)) was significantly associated only with white cell counts, nitrogen dioxide (NO(2)) with platelet counts and fibrinogen, and ozone with none of the outcomes. In two-pollutant models, PM(10) remained a significant predictor of white cell counts controlling for SO(2) but not vice versa. PM(10) was marginally significant in a model for platelet counts with NO(2), and the sign of the NO(2) coefficient was reversed. These results were stable with control for indoor exposures (wood stoves, environmental tobacco smoke, gas stoves, fireplaces), dietary risk factors (saturated fat, alcohol, caffeine intake, n-3 fatty acids), and serum cholesterol. The magnitude of the effects are modest [e.g., 13 microg/dL fibrinogen for an interquartile range (IQR) change in PM(subscript)10(/subscript), 95% confidence interval (CI) 4.6 22.1 mg/dL]. However, the odds ratio of being in the top 10% of fibrinogen for the same IQR change was 1.77 (95% CI 1.26-2.49). These effects provide considerable biologic plausibility to the mortality studies. PM(10), but not gaseous air pollutants, is associated with blood markers of cardiovascular risk, and this may explain epidemiologic associations with early deaths. PMID- 11427391 TI - Assessing the health benefits of urban air pollution reductions associated with climate change mitigation (2000-2020): Santiago, Sao Paulo, Mexico City, and New York City. AB - To investigate the potential local health benefits of adopting greenhouse gas (GHG) mitigation policies, we develop scenarios of GHG mitigation for Mexico City, Mexico; Santiago, Chile; Sao Paulo, Brazil; and New York, New York, USA using air pollution health impact factors appropriate to each city. We estimate that the adoption of readily available technologies to lessen fossil fuel emissions over the next two decades in these four cities alone will reduce particulate matter and ozone and avoid approximately 64,000 (95% confidence interval [CI] 18,000-116,000) premature deaths (including infant deaths), 65,000 (95% CI 22,000-108,000) chronic bronchitis cases, and 46 million (95% CI 35-58 million) person-days of work loss or other restricted activity. These findings illustrate that GHG mitigation can provide considerable local air pollution related public health benefits to countries that choose to abate GHG emissions by reducing fossil fuel combustion. PMID- 11427392 TI - Farmworker pesticide exposure and community-based participatory research: rationale and practical applications. AB - The consequences of agricultural pesticide exposure continue to be major environmental health problems in rural communities. Community-based participatory research (CBPR) is an important approach to redressing health disparities resulting from environmental causes. In this article we introduce a collection of articles that describe projects using CBPR to address the health disparities resulting from pesticide exposure in agricultural communities, particularly the communities of migrant and seasonal farmworkers. The articles in this collection are based on a workshop convened at the 1999 American Public Health Association meeting. The goals in presenting this collection are to provide those endeavoring to initiate CBPR projects needed information, guidelines, and procedures to improve the quality of the CBPR experience; to increase the scientific validity of CBPR projects; and to reduce the potential difficulties and stress of these collaborations. In this introduction we discuss the context in which these projects operate, summarizing background information about farmworkers in the United States, what is known about farmworker pesticide exposure, and the concept of community-based participatory research. Finally, the articles in this collection are summarized, and major themes common to successful CBPR projects are identified. These common features are taking the time to interact with the community, using multiple approaches to engage the different parts of the community, understanding different participants often have different goals, appreciating each group's strengths, valuing community knowledge, and being flexible and creative in conducting research. The final article in this collection describes the translational research program at the National Institute of Environmental Health Sciences (NIEHS) highlighting activities pertinent to the health of rural communities, giving an overview of NIEHS-supported projects addressing health concerns of Native Americans and rural African-American communities in addition to farmworkers, and discussing future plans for CBPR at NIEHS. PMID- 11427393 TI - Something for everyone? A community and academic partnership to address farmworker pesticide exposure in North Carolina. AB - Partnerships between academic researchers and community organizations are frequently formed to address environmental health concerns in underserved communities. Although such participatory approaches to research combine valuable assets of both partners, they are often difficult to maintain. We describe a partnership formed to investigate migrant and seasonal farmworker exposure to pesticides in North Carolina and to develop effective interventions to reduce exposure. North Carolina ranks fifth in the United States in the number of farmworkers; most are from Mexico, and a significant minority come to the United States on work contracts. Several barriers to establishing effective collaboration were recognized in this partnership, including stereotypes, cultural differences, competing demands for time and attention, and differences in orientation to power structures. To overcome these barriers, members of the partnership took actions in three domains: clarifying the different goals of each partner, operationalizing a model of participation that could involve many different community segments developing cultural sensitivity. By taking these actions, the work of the partnership was accomplished in ways that met the criteria for success of both academic researchers and community members. This approach can be used by others to develop collaborative relationships to investigate environmental health issues within a community-based participatory framework. PMID- 11427394 TI - Identifying constituents to participate in a project to control pesticide exposure in children of farmworkers. AB - Farmers in Washington State use pesticides to control harmful pests that might interfere with the quality of their products. Farmworkers, who are primarily responsible for thinning, harvesting, and other agricultural work, are often exposed to these pesticides and take home pesticide residues on their clothing, shoes, and skin, potentially exposing children in the household to pesticides. We designed a project to reduce children's exposure by using a community organization model. To better understand the community views regarding pesticide exposure, we conducted a qualitative community analysis. Two methods of data gathering were used to collect information. Individual interviews were conducted with some respondents, and small group discussions were held with others. Analysis indicated wide disparity among involved groups in their views on pesticides; however, a number of themes common to the majority of the constituents were identified and discussed with a community planning group. Because of the contention around pesticides, the group recommended every constituent participate in decision making. The group noted it was important to emphasize that the research project could provide scientific information on pesticide exposure to everyone in the Lower Yakima Valley. Our study demonstrates both the widely varying views of different constituents around the issue of pesticide exposure and common themes that can form the basis for collaboration and consensus on approaching the issue. PMID- 11427395 TI - The Oregon migrant farmworker community: an evolving model for participatory research. AB - Migrant farmworker communities present distinct challenges that require new approaches for community participation in research. In the State of Oregon an agency that advocates for the migrant farmworker community has collaborated successfully with university researchers to implement a research program directed to reducing pesticide exposures among the children of migrant farmworkers. The research process has included both qualitative research methods with members of the community and quantitative approaches to measure pesticide dust residues in homes, biomarkers of pesticide exposure, and effects on health. A committee of university and community stakeholders advises the research. Evaluative processes have been initiated to assess the effectiveness of the participatory model used in this project. The components of the preliminary process evaluation and results are presented. Evaluative data show that researchers and community members differ on perceptions of community involvement and the extent to which communication problems have been resolved between the two groups. Suggestions for improved community involvement and communication are given. PMID- 11427396 TI - Recruiting a community sample in collaboration with farmworkers. AB - Few studies have examined health effects of pesticides in farmworkers, possibly because researchers perceive this population to be relatively inaccessible. We conducted an epidemiologic study of health effects among farmworkers in two towns in central Florida--Apopka and Pierson. Apopka is a suburb of Orlando with a diffuse farmworker community working in many crops, whereas Pierson is a small rural town with a tightly knit farmworker community working mainly in ferns. We collaborated with the Farmworker Association of Florida, a grassroots organization representing 6,700 farmworker families. We identified potential participants using membership lists of the Community Trust Federal Credit Union. Members of the Farmworker Association served as recruiters for the study, locating randomly selected Credit Union members and administering a screening interview to determine eligibility. In Apopka 90% of contacted workers were screened, and 79% of eligible workers participated in the study; corresponding proportions in Pierson were 94 and 85%. Farmworkers who had worked for 6-15 years and those who worked in a defined type of agriculture (nursery, citrus, or ferns) were more likely to enroll than others. Thus, while the response rate was good for a multistage recruiting process, study participants had a slightly different work history from those who chose not to enroll. We conclude that it is possible to conduct a study of health outcomes in farmworkers with a defined population and good response rates. Collaboration with the community is essential to the success of such a project, and community characteristics can affect response rates. PMID- 11427397 TI - Implementing a community-based social marketing project to improve agricultural worker health. AB - The Together for Agricultural Safety project is a community-based social marketing project working to reduce the adverse health effects of pesticide exposure among fernery and nursery workers in Florida. In 3 years, the collaboration between university and community researchers has embodied many of the principles of community-based research while completing multiple stages of formative data collection required for a social marketing project. This hybrid approach to developing a health intervention for a minority community has been successful in its early stages because the community partners are organized, empowered, and motivated to execute research activities with the assistance of academic partners. However, this work has also been labor intensive and costly. This article describes the lessons learned by project partners and considers the limitations of this approach for agricultural health research. PMID- 11427398 TI - Commitment of the National Institute of Environmental Health Sciences to community-based participatory research for rural health. AB - The National Institute of Environmental Health Sciences (NIEHS) is the leading biomedical research institution in the United States whose mission is to support research that seeks to understand how environmental exposures affect human health. NIEHS possesses a longstanding interest in the health effects of agrochemical and other environmental exposures in rural America, including pesticides, to farmers and their families and to migrant farmworkers and their families. In recent years, NIEHS has begun augmenting traditional basic science investigations with innovative programs that translate findings from the laboratory to affected populations. It is through community-based participatory research that NIEHS strives to advance the public health field by fostering the development of culturally relevant interventions that will reduce exposures to environmental contaminants and the risk of environmentally induced disease. In this article, we describe the translational research program at NIEHS as it relates to the NIEHS mission and highlight activities pertinent to the health of rural communities, especially underserved populations. We provide an overview of NIEHS-supported projects addressing health concerns of Native American and rural African-American communities in addition to farmworkers. We conclude with a discussion of future plans for community-based participatory research at NIEHS. PMID- 11427399 TI - Delta-aminolevulinic acid dehydratase genotype and lead toxicity: a HuGE review. AB - The ALAD gene (chromosome 9q34) codes for delta-aminolevulinic acid dehydratase (ALAD) (E.C. 4.2.1.24). ALAD catalyzes the second step of heme synthesis and is polymorphic. The ALAD G177C polymorphism yields two codominant alleles, ALAD-1 and ALAD-2, and it has been implicated in susceptibility to lead toxicity. Genotype frequencies vary by geography and race. The rarer ALAD-2 allele has been associated with high blood lead levels and has been thought to increase the risk of lead toxicity by generating a protein that binds lead more tightly than the ALAD-1 protein. Other evidence suggests that ALAD-2 may confer resistance to the harmful effects of lead by sequestering lead, making it unavailable for pathophysiologic participation. Recent studies have shown that individuals who are homozygous for the ALAD-1 allele have higher cortical bone lead levels; this implies that they may have a greater body lead burden and may be at higher risk of the long-term effects of lead. Individuals exposed to lead in occupational settings have been the most frequent subjects of study. Genotype selection bias may limit inferences from these studies. No firm evidence exists for an association between ALAD genotype and susceptibility to lead toxicity at background exposure levels; therefore, population testing for the ALAD polymorphism is not justified. PMID- 11427400 TI - Risk factors for neonatal seizures: a population-based study, Harris County, Texas, 1992-1994. AB - Risk factors for neonatal seizures were evaluated in 116,048 infants born between 1992 and 1994 to residents of Harris County, Texas; 207 of these infants were diagnosed with clinical neonatal seizures. Information was obtained from the infant's birth certificate to assess the relation between seizures and birth weight, gender, ethnicity, place of birth, mother's age, method of delivery, parity, and multiple births. These factors were evaluated by univariate and multivariate analysis using logistic regression. For preterm infants, a birth weight of <1,500 g was the strongest risk factor (relative risk (RR) = 9.1, 95% confidence interval (CI): 4.7, 17.5), followed by birth in a private/university hospital (RR = 2.8, 95% CI: 1.5, 5.0) and male gender (RR = 1.8, 95% CI: 1.0, 3.4). For term infants, significant risk factors included birth by cesarean section (RR = 2.2, 95% CI: 1.5, 3.2), small birth weight for gestational age (RR = 1.9, 95% CI: 1.2, 2.9), birth in a private/university hospital (RR = 1.8, 95% CI: 1.1, 3.0), and maternal age of 18-24 compared with 25-29 years (RR = 1.6, 95% CI: 1.1, 2.3). Birth by assisted vaginal delivery and primiparity were marginally significant for term infants. Birth weight is a significant risk factor for neonatal seizures. The role of perinatal complications warrants further evaluation. PMID- 11427401 TI - Tracking of body size from birth to late adolescence: contributions of birth length, birth weight, duration of gestation, parents' body size, and twinship. AB - Birth weight has correlated positively with adult body mass index (BMI), but rarely have birth length, duration of gestation, or parents' body size been taken into account. The authors examined tracking of birth length and weight, adjusted for gestational age, to late adolescence, with special reference to parents' height and BMI. Longitudinal information from a nationally representative sample of Finnish twin adolescents (birth cohorts 1975-1979) and their parents was collected via questionnaires mailed when the twins were aged 16 years (n = 4,376; 2,062 males, 2,314 females) and 18 years (n = 3,917; 1,742 males, 2,175 females). The twins showed significant tracking of body size from birth to late adolescence, which was greatly influenced by their parents' body size. Height in adolescence was predicted by length and weight at birth and by parents' height, whereas BMI was predicted by birth weight and parents' BMI. An especially high risk for overweight was found for subjects of average length but a high weight at birth. These findings suggest that the intrauterine period has enduring effects on later body size but leave unresolved whether these effects are genetic or environmental. PMID- 11427402 TI - Risk factors for the development of low back pain in adolescence. AB - A previous history and earlier onset of low back pain are associated with chronic low back pain in adults, implying that prevention in adolescence may have a positive impact in adulthood. The study objectives were to determine the incidence of low back pain in a cohort of adolescents and to ascertain risk factors. A cohort of 502 high school students in Montreal, Canada, was evaluated during 1995-1996 at three separate times, 6 months apart. The outcome was low back pain occurrence at a frequency of at least once a week in the previous 6 months. Of the 377 adolescents who did not complain of low back pain at the initial evaluation, 65 developed low back pain over the year (cumulative incidence, 17 percent). Risk factors associated with development of low back pain were high growth (odds ratio = 3.09; 95 percent confidence interval (CI): 1.53, 6.01), smoking (odds ratio = 2.20; 95% CI: 1.38, 3.50), tight quadriceps femoris (odds ratio = 1.02; 95% CI: 1.00, 1.05), tight hamstrings (odds ratio = 1.04; 95% CI: 1.01, 1.06), and working during the school year (odds ratio = 1.33; 95% CI: 1.03, 1.71). Modifying such risk factors as smoking and poor leg flexibility may potentially serve to prevent the development of low back pain in adolescents. PMID- 11427403 TI - Breastfeeding and reduced risk of breast cancer in an Icelandic cohort study. AB - Case-control studies on the association between breastfeeding and the subsequent risk of breast cancer have given inconsistent results. To date, only two cohort studies have been reported on this subject. The present nested case-control study uses data from an Icelandic cohort of 80,219 women visiting a Cancer Detection Clinic that offers population-based cervical and breast cancer screening, in the years 1979-1995. The 993 parous cases were aged 26-90 years at diagnosis, with 9,729 parous controls individually matched on birth year, vital status at case diagnosis, and age when giving information on several potential risk factors for breast cancer. Using conditional logistic regression and confining the analysis to the 84 cases who were <40 years at diagnosis, an inverse association was evident between total duration of breastfeeding and breast cancer, with the adjusted odds ratio = 0.77 per 6 months' increase in duration of breastfeeding (95% confidence interval: 0.59, 1.00), whereas for the remainder of the women, a much weaker trend was observed. Ever lactating was associated with decreased risk, with the adjusted odds ratio = 0.33 (95% confidence interval: 0.19, 0.56) for women diagnosed at all ages. This is the first cohort study to indicate a negative association between breastfeeding and breast cancer. PMID- 11427404 TI - Luteinizing hormone, its beta-subunit variant, and epithelial ovarian cancer: the gonadotropin hypothesis revisited. AB - The gonadotropin hypothesis postulates that excessive gonadotropin stimulation results in increased proliferation and subsequent malignant transformation of ovarian epithelium. The authors evaluated this hypothesis by analyzing the association between serum levels of wild-type luteinizing hormone (LH) and ovarian cancer risk. They also examined the relation between a variant of LH containing two missense point mutations (Trp(8)Arg and Ile(15)Thr) in its beta subunit and ovarian cancer risk. Fifty-eight cases of epithelial ovarian cancer and 116 controls matched on age, menopausal status, and date of blood donation were included in a case-control study nested within the New York University Women's Health Study, a prospective cohort enrolled between 1985 and 1991 in New York City. Wild-type serum levels and variant LH status were determined by immunofluorometric assays in which monoclonal antibodies specific for wild-type and variant LH were used. Compared with women in the lowest tertile of wild-type LH, women in the highest tertile had a lower risk of ovarian cancer, after adjustment for potential confounders (odds ratio = 0.42, 95% confidence interval: 0.09, 2.09). Women heterozygous for variant LH were not at increased risk (adjusted odds ratio = 0.95, 95% confidence interval: 0.27, 3.34). The results suggest that neither wild-type LH levels nor variant LH status is associated with increased risk of epithelial ovarian cancer. PMID- 11427405 TI - Leisure-time physical activity levels and changes in relation to risk of hip fracture in men and women. AB - The authors prospectively studied the effect of leisure-time physical activity level on hip fracture risk along with the influence of within-subject changes in activity levels, while taking possible confounding by other health behaviors and poor health into account. Analyses were based on pooled data from three population studies conducted in Copenhagen, Denmark. Among 13,183 women and 17,045 men, 1,121 first hip fractures were identified during follow-up. In comparison with being sedentary, the relative risk (RR) of hip fracture associated with being moderately physically active 2-4 hours per week was 0.72 (95% confidence interval (CI): 0.59, 0.89) in women and 0.75 (95% CI: 0.55, 1.03) in men after adjustment for confounders. Being in the most active leisure activity category did not decrease the risk of hip fracture further. Adjustment for poor health affected the risk estimates only modestly. Subjects who, during follow-up, reduced their physical activity level from the highest or the intermediate activity level to a sedentary level had a higher risk of hip fracture than did those who remained moderately physically active at the intermediate level (multivariate adjusted RR = 2.19, 95% CI: 1.00, 4.84 and RR = 1.89, 95% CI: 1.21, 2.95, for reduction from the highest and intermediate levels, respectively). There was no evidence of a fracture-protective effect from increasing physical activity. In conclusion, moderate levels of physical activity appear to provide protection against later hip fracture. Decline in the physical activity level over time is an important risk factor for hip fracture. PMID- 11427406 TI - Cigarette smoking and incidence of multiple sclerosis. AB - Experimental data suggest that cigarette smoking may play a role in the development of multiple sclerosis (MS), but epidemiologic studies have been small and inconclusive. The authors assessed the association between MS incidence and smoking in two cohort studies of US women, the Nurses' Health Study (121,700 women aged 30-55 years at baseline in 1976) and the Nurses' Health Study II (116,671 women aged 25-42 years at baseline in 1989). Smoking history was assessed at baseline and updated on biennial questionnaires. A total of 315 definite or probable cases of MS were documented. Compared with that for women who never smoked, the relative incidence rate was 1.6 (95% confidence interval: 1.2, 2.1) among current smokers and 1.2 (95% confidence interval: 0.9, 1.6) among past smokers after adjustment for age, latitude, and ancestry. The relative rate increased significantly with cumulative exposure to smoking (p for trend < 0.05), from 1.1 (95% confidence interval: 0.8, 1.6) for 1-9 pack-years to 1.5 (95% confidence interval: 1.2, 2.1) for 10-24 pack-years and 1.7 (95% confidence interval: 1.2, 2.4) for 25 or more pack-years. Similar results were obtained after adjustment for other potential confounding factors. Although these prospective results do not prove a cause-and-effect relation, they suggest that smoking is associated with an increased risk of MS. PMID- 11427407 TI - Prognostic value of cross-sectional anthropometric indices on short-term risk of mortality in human immunodeficiency virus-infected adults in Abidjan, Cote d'Ivoire. AB - In sub-Saharan Africa where weight loss is very difficult to estimate, cross sectional anthropometric indicators could be useful to predict human immunodeficiency virus (HIV)-associated mortality. The study objective was to look for threshold values of baseline body mass index, arm muscle circumference, and fat mass to predict the risk of death in HIV-infected adults included in a 1996-1998 trial of early cotrimoxazole chemoprophylaxis in Abidjan, Cote d'Ivoire (COTRIMO-CI-ANRS 059 trial). The authors graphically determined if consecutive anthropometric categories with the closest hazards ratios of the risk of death could be clustered to obtain a unique threshold that distinctly separated two categories. When the threshold values were determined, the authors estimated the hazards ratio of mortality of this two-category model. A significant increase of mortality was observed for a body mass index of < or =20.3 in men (hazards ratio = 2.6; 95% confidence interval (CI): 1.4, 5.0) and of < or =18.5 in women (hazards ratio = 2.2; 95% CI: 1.05, 4.5) and for a fat mass of < or =6% in men (hazards ratio = 4.6; 95% CI: 2.3, 9.4) and of < or =18% in women (hazards ratio = 2.4; 95% CI: 1.2, 4.9). No simple threshold could be identified for arm muscle circumference. In Cote d'Ivoire where chemoprophylaxis of opportunistic infections has recently been recommended to be widely initiated on clinical criteria, such thresholds may help to screen patients with higher risks of mortality. PMID- 11427408 TI - Interpreting results from trials of pneumococcal conjugate vaccines: a statistical test for detecting vaccine-induced increases in carriage of nonvaccine serotypes. AB - Conjugate vaccines against Streptococcus pneumoniae (pneumococcus) protect against nasopharyngeal carriage of serotypes included in the vaccine. However, in several clinical trials, vaccinees have shown increased carriage of nonvaccine serotypes of pneumococcus. These increases may be due to serotype replacement, if vaccine-induced protection against carriage of vaccine serotypes increases susceptibility to carriage of nonvaccine serotypes. Alternatively, observed increases may be an artifact of "unmasking," in which nonvaccine serotypes are more readily detected among vaccinees than among controls because vaccine serotypes are not present. In this paper, a statistical test for distinguishing serotype replacement from unmasking is described. The test attempts to reject a null model of unmasking alone; serotype replacement is inferred if the observed increase in detectable nonvaccine serotype carriage among vaccinees is significantly greater than that expected under the null model. Significance is assessed using the Bayesian "posterior predictive p value" as modified by Robins et al. (J Am Stat Assoc 2000;95:1143-56). Analysis of data from a South African trial suggests that replacement may have occurred in the study, but results do not reach the conventional level of significance in rejecting the null hypothesis of unmasking (p = 0.074). The author performs sensitivity analyses for the prior and for unmeasured confounding by differences in susceptibility to pneumococcus carriage. The implications of the findings and the assumptions and limitations of this technique are then discussed. PMID- 11427409 TI - Immunological effects of interleukin-2 therapy in human immunodeficiency virus positive subjects. PMID- 11427410 TI - Celiac disease-associated autoimmune endocrinopathies. AB - Celiac disease (CD) is an autoimmune disorder induced by gluten intake in genetically susceptible individuals. It is characterized by the presence of serum antibodies to endomysium, reticulin, gliadin, and tissue transglutaminase. The incidence of CD in various autoimmune disorders is increased 10- to 30-fold in comparison to the general population, although in many cases CD is clinically asymptomatic or silent. The identification of such cases with CD is important since it may help in the control of type I diabetes or endocrine functions in general, as well as in the prevention of long-term complications of CD, such as lymphoma. It is believed that CD may predispose an individual to other autoimmune disorders such as type I diabetes, autoimmune thyroid, and other endocrine diseases and that gluten may be a possible trigger. The onset of type I diabetes at an early age in patients with CD, compared to non-CD, and the prevention or delay in onset of diabetes by gluten-free diet in genetically predisposed individuals substantiates this antigen trigger hypothesis. Early identification of CD patients in highly susceptible population may result in the treatment of subclinical CD and improved control of associated disorders. PMID- 11427411 TI - Antituberculosis treatment: increasing evidence for drug effects on innate cellular immunity. PMID- 11427412 TI - Comparison of hepatitis C viral loads in patients with or without human immunodeficiency virus. AB - A better understanding of how human immunodeficiency virus (HIV) coinfection affects the course of hepatitis C virus (HCV) infection is required to select patients with HIV who would benefit from current HCV therapy. Between June 1996 and March 2000, HCV RNA levels were quantified for 1,279 patients at the Louisiana State University Health Sciences Center; 28 of these patients were coinfected with HIV. HCV loads were quantified by the Bayer branched-DNA assay with a lower limit of detection of 0.2 Meq/ml. We compared the median HCV RNA levels of for patients coinfected with HIV and HCV and patients infected only with HCV who were in the same age range (23 to 55 years). The median HCV load for the 28 patients coinfected with HCV and HIV (17.8 Meq/ml) was significantly greater (P < 0.05) than that for similarly aged patients infected only with HCV (6.1 Meq/ml). The HCV load did not correlate with age or sex for either group of patients. A significant (R = -0.4; P < 0.05) negative correlation was observed between HCV load and CD4 count in the coinfected group, for whom the CD4 counts at the time of HCV load analysis ranged from 6 to 1,773/mm(3). The increased HCV load in patients coinfected with HCV and HIV compared to that in patients infected only with HCV and the inverse relationship of the HCV load to the CD4 count indicate that immunosuppression results in decreased control of HCV replication. In addition, we report significantly higher HCV loads among coinfected African Americans than Caucasians. PMID- 11427413 TI - Induction by a lactic acid bacterium of a population of CD4(+) T cells with low proliferative capacity that produce transforming growth factor beta and interleukin-10. AB - We investigated whether certain strains of lactic acid bacteria (LAB) could antagonize specific T-helper functions in vitro and thus have the potential to prevent inflammatory intestinal immunopathologies. All strains tested induced various levels of both interleukin-12 (IL-12) and IL-10 in murine splenocytes. In particular, Lactobacillus paracasei (strain NCC2461) induced the highest levels of these cytokines. Since IL-12 and IL-10 have the potential to induce and suppress Th1 functions, respectively, we addressed the impact of this bacterium on the outcome of CD4(+) T-cell differentiation. For this purpose, bacteria were added to mixed lymphocyte cultures where CD4(+) T-cells from naive BALB/c mice were stimulated weekly in the presence of irradiated allogeneic splenocytes. In these cultures, L. paracasei NCC2461 strongly inhibited the proliferative activity of CD4(+) T cells in a dose-dependent fashion. This was accompanied by a marked decrease of both Th1 and Th2 effector cytokines, including gamma interferon, IL-4, and IL-5. In contrast, IL-10 was maintained and transforming growth factor beta (TGF-beta) was markedly induced in a dose-dependent manner. The bacteria were not cytotoxic, because cell viability was not affected after two rounds of stimulation. Thus, unidentified bacterial components from L. paracasei NCC2461 induced the development of a population of CD4(+) T cells with low proliferative capacity that produced TGF-beta and IL-10, reminiscent of previously described subsets of regulatory cells implicated in oral tolerance and gut homeostasis. PMID- 11427414 TI - In vitro hydroxyurea decreases Th1 cell-mediated immunity. AB - Hydroxyurea (HU) is used in the treatment of hematologic disorders and is sometimes added to antiretroviral combination therapy to potentiate human immunodeficiency virus (HIV) suppression. However, HU has toxic effects on rapidly dividing cells, including the effectors of the immune response. To determine whether HU affects specific T-cell responses, we measured lymphocyte proliferation and cytokine production in response to microbial antigen and mitogen stimulation in the presence of added HU (10 to 1,000 microM). HU treatment of peripheral blood mononuclear cells obtained from HIV-infected patients and uninfected controls decreased lymphocyte proliferation and gamma interferon production compared with untreated cells. Interleukin-2 (IL-2) and IL 10 production was not affected by HU. The HU-mediated decrease of lymphocyte proliferation was similar in peripheral blood mononuclear cells from HIV-infected patients and from uninfected controls. The inhibitory effect of HU required continuous exposure to the drug and could be reverted by washing the drug out of the culture environment. These findings suggest that HU-containing therapeutic regimens might decrease Th1-cell-mediated immune responses in vivo. PMID- 11427415 TI - Biological response modifier activity of an exopolysaccharide from Paenibacillus jamilae CP-7. AB - An extracellular polysaccharide was purified from culture supernatants of Paenibacillus jamilae CP-7, a gram-positive bacillus that was isolated from compost prepared with olive mill wastewaters. The extracellular polysaccharide was produced under aerobic conditions in a medium containing olive mill wastewaters (80% [vol/vol]). This exopolymer had a low level of acute toxicity when it is administered to BALB/c mice by the intraperitoneal route. Interesting immunomodulatory effects were detected when mice were given 10 mg of exopolysaccharide per kg of body weight; the proliferative responses of splenocytes to B-cell and T-cell mitogens were suppressed, the in vitro levels of production of gamma interferon and granulocyte-macrophage colony-stimulating factor by unstimulated and lipopolysaccharide-stimulated splenocytes were enhanced, and the levels of resistance to the intracellular pathogen Listeria monocytogenes was increased in mice. Also, the exopolysaccharide was able to induce lymphocyte proliferation in vitro. We conclude that P. jamilae produces an exopolysaccharide with interesting immunomodulatory properties. PMID- 11427416 TI - Isogenic strain of Escherichia coli O157:H7 that has lost both Shiga toxin 1 and 2 genes. AB - An Escherichia coli O157:H7 strain isolated from a patient with hemorrhagic colitis was found to exhibit two slightly different colony morphology types on differential medium. Each morphological type, designated TT12A and TT12B, was isolated, and serological testing using various assays confirmed that both strains carried the O157 and the H7 antigens. Biochemical testing showed that the strains had identical profiles on AP120E analysis and, like typical O157:H7 strains, did not ferment sorbitol or exhibit beta-glucuronidase activity. Analysis with a multiplex PCR assay showed that TT12B did not carry the gene for either Shiga toxin 1 (Stx1) or Stx2, whereas these genes were present in TT12A and the toxins were produced. Apart from that, both strains carried the +93 gusA mutation, the cluster I ehxA gene for enterohemolysin, and the eae gene for gamma intimin, which are all characteristics of the O157:H7 serotype. Phenotypic assays confirmed that both strains exhibited enterohemolysin activity and the attachment and effacing lesion on HeLa cells. Multilocus enzyme electrophoresis analysis showed that the strains are closely related genetically and belong in the same clonal group. Pulsed-field gel electrophoresis (PFGE) typing of XbaI-digested genomic DNA revealed that the two strains differed by two bands but shared 90% similarity and clustered in the same clade. All other non-Stx-producing O157:H7 strains examined clustered in a major clade that was distinct from that of Stx producing O157:H7 strains. The findings that TT12B was identical to TT12A, except for Stx production, and its PFGE profile is also more closely related to that of Stx-producing O157:H7 strains suggest that TT12B was derived from TT12A by the loss of both stx genes. PMID- 11427417 TI - Posttreatment follow-up of Helicobacter pylori infection using a stool antigen immunoassay. AB - The Helicobacter pylori stool antigen enzyme immunoassay (HpSA) was evaluated during posttreatment follow-up of patients in a country with a very high prevalence of H. pylori infection. From among 273 dyspeptic individuals (18 to 55 years) initially recruited from a shantytown in Lima, Peru, 238 participants who met the inclusion criteria and were suspected to be H. pylori positive based on (14)C urea breath test (UBT) results underwent endoscopy. Participants with endoscopy-proven infections received standard eradication therapy and were monitored by UBT and HpSA at 1 month following treatment and at 3-month intervals for 9 months posttreatment. A second endoscopy was performed if UBT results showed evidence of treatment failure or H. pylori recurrence. Biopsy results were considered the "gold standard" in all analyses. Among patients who underwent endoscopy, HpSA had a pretreatment sensitivity of 93%. Two-hundred thirty patients completed the treatment regimen, of whom 201 (93%) were considered to have had successful treatment outcomes based on a negative follow-up UBT. Thirty two patients with UBT-defined treatment failures or H. pylori recurrences at any point during the 9-month follow-up underwent a second endoscopy. In the posttreatment setting, HpSA had an overall sensitivity of 73% and a specificity of 67%. Agreement between UBT and HpSA diminished throughout the follow-up. Among 14 participants in whom HpSA remained positive at 1 month following treatment despite UBT evidence of treatment success, 12 (86%) became HpSA negative within 3 months posttreatment. Although this study confirmed the validity of the HpSA in the initial assessment of dyspeptic patients, the test demonstrated a reduced overall accuracy in the detection of treatment failures and H. pylori recurrences during 9 months of posttreatment follow-up. Furthermore, in some patients it may take up to 3 months after successful eradication for antigen shedding to diminish to levels within the negative HpSA range. PMID- 11427418 TI - Standardized method of measuring acanthamoeba antibodies in sera from healthy human subjects. AB - Acanthamoeba species can cause serious, debilitating, and sometimes life threatening infections. Three groups have been identified using morphological and immunological comparisons. Previous serological studies have utilized a variety of antigen preparations and assay methods and reported disparate (3 to 100%) results. This study was designed to (i) optimize an enzyme-linked immunosorbent assay for detecting serum antibodies to each of the Acanthamoeba serogroups and (ii) test 55 healthy individuals for specific immunoglobulin G reactivity. The highest signal-to-background ratio was found when 3,000 fixed, intact trophozoites per well were used with a 1:10 serum dilution. Sera yielding optical densities of <0.25 against all three Acanthamoeba serogroups were used to define the cutoff for positive results. The highest background reactivity with these sera was seen with Acanthamoeba polyphaga (serogroup 2), followed by Acanthamoeba culbertsoni (serogroup 3) and Acanthamoeba astronyxis (serogroup 1). Of 55 subjects tested, the highest number of positive results was seen with A. polyphaga (81.8%), followed by A. astronyxis (52.8%) and A. culbertsoni (40%). Seven serum samples (12.7%) were negative for all three Acanthamoeba serogroups, 16 (29.1%) were positive for one serogroup only, 16 were positive for two serogroups, and 16 reacted to all three serogroups. Further analysis showed no significant associations between serogroup reactivity and age or gender. However, some ethnic differences were noted, especially with A. polyphaga antigens. In that case, serum samples from Hispanic subjects were 14.5 times less likely to be positive (P = 0.0025) and had lower mean absorbance values (P = 0.047) than those from Caucasian subjects. Overall, these data suggest that Acanthamoeba colonization or infection is more common than previously thought. Mild or asymptomatic infections may contribute to the observed serum reactivities. PMID- 11427419 TI - Helicobacter pylori intrafamilial infections: change in source of infection of a child from father to mother after eradication therapy. AB - Biopsy specimens of the antrum and corpus were obtained from four Helicobacter pylori-infected members of a family and from the same boy (son 1) in whom the infection reappeared after simultaneous successful eradication treatment of three family members, excluding the mother. A total of 18 to 60 H. pylori isolates were obtained from each specimen and subjected to rRNA gene restriction pattern analysis. The father's isolates and the initial isolates from son 1 showed the same HindIII type, which was divided into three HaeIII subtypes. Isolates from the mother and a brother (son 2) and posttreatment isolates from son 1 showed a distinct HindIII type (with one minor subtype), which was divided into six HaeIII subtypes. All subtypes of the initial isolates from son 1 were present in the father's isolates, and all subtypes of the posttreatment isolates from son 1 were present in the mother's isolates but not in son 2's. Electron microscopic analysis of the biopsy specimens demonstrated extremely high levels of H. pylori colonization in the father's gastric mucosa. H. pylori adherence with a ruffle formation was also demonstrated. The findings suggest that son 1 was infected initially with the H. pylori strain of the father and son 2 was infected with the H. pylori strain of the mother and that after eradication therapy son 1 was reinfected with the H. pylori strain of the mother, who did not undergo eradication therapy. PMID- 11427420 TI - Immune function in healthy inner-city children. AB - The importance of investigating immunity in healthy children has been underscored in the last few years by studies of the immune pathology of childhood illnesses, including human immunodeficiency virus. This study reports both ennumerative and functional immune measures in healthy inner city children. A total of 152 of 207 children studied were completely heathy at the time of venipuncture and were included in this study. Laboratory immune batteries were completed (or begun) the same day as venipuncture. Relationships between age, gender, ethnicity, and immunity were then analyzed. We found that gender predicted both the absolute number and the percentage of T cells and helper cells and the percentage of natural killer cells. Total leukocyte counts and percentages of lymphocytes and granulocytes were related to ethnicity, as was the response to mitogen stimulation (concanavalin A and pokeweed mitogen) and phagocytic ability. In conclusion, age, gender, and ethnicity factors were found to contribute to differences in various immune measures in children and require further investigation. PMID- 11427421 TI - Cross-reactivity of Epstein-Barr virus-specific immunoglobulin M antibodies with cytomegalovirus antigens containing glycine homopolymers. AB - Timely and reliable detection of acute primary human cytomegalovirus (HCMV) infection is important in prenatal screening programs and for differential diagnosis of infectious mononucleosis-like disease. Enzyme-linked immunosorbent assays (ELISAs) based on HCMV proteins enable the sensitive detection of immunoglobulin M (IgM) antibodies during primary infection. However, concerns have been raised about possible cross-reactivities of the HCMV antigens used for the design of such ELISAs with IgM antibodies induced by Epstein-Barr Virus (EBV). In this study we investigated whether IgM antibodies generated during acute EBV infection reacted with recombinant HCMV antigens. Serum samples from patients with primary EBV infection frequently scored positive when tested in different HCMV IgM ELISAs, irrespective of whether conventional or recombinant antigens were used for the design of the HCMV IgM assays. Such cross-reactive IgM antibodies were found to be directed against short glycine-rich motifs contained within the nonstructural HCMV proteins pUL44 and pUL57. Further analyses revealed that these glycine-rich motifs were major antigenic domains for IgM antibodies induced during HCMV infection. Their deletion from recombinant proteins abrogated reactivity with IgM synthesized during HCMV infection. EBV-induced IgM antibodies that reacted with HCMV antigens showed similar kinetics of reactivity in HCMV- or EBV-specific assays in the course of primary EBV infection, indicating that the two populations of antibodies were highly overlapping. The results demonstrate that primary EBV infection leads to the induction of IgM antibodies that specifically bind to widely used diagnostic antigens of HCMV. This has to be considered in the interpretation of HCMV-specific IgM assays. PMID- 11427422 TI - Role of Nijmegen breakage syndrome protein in specific T-lymphocyte activation pathways. AB - Nijmegen breakage syndrome (NBS) is a genetic disorder characterized by immunodeficiency, microcephaly, and "bird-like" facies. NBS shares some clinical features with ataxia telangiectasia (AT), including increased sensitivity to ionizing radiation, increased spontaneous and induced chromosome fragility, and strong predisposition to lymphoid cancers. The mutated gene that results in NBS codes for a novel double-stranded DNA break repair protein, named nibrin. In the present work, a Spanish NBS patient was extensively characterized at the immunological and the molecular DNA levels. He showed low CD3(+)-cell numbers and an abnormal low CD4(+) naive cell/CD4(+) memory cell ratio, previously described in AT patients and also described in the present report in the NBS patient. The proliferative response of peripheral blood lymphocytes in vitro to mitogens is deficient in NBS patients, but the possible link among NBS mutations and the abnormal immune response is still unknown. PMID- 11427423 TI - Enhanced antigen-specific delayed-type hypersensitivity and immunoglobulin G2b responses after oral administration of viable Lactobacillus casei YIT9029 in Wistar and Brown Norway rats. AB - In this study, the effects of orally administered viable Lactobacillus casei Shirota strain YIT9029 on the immunity parameters of Wistar and Brown Norway rats were examined. For this purpose, we used the Trichinella spiralis host resistance model. Two weeks before and during T. spiralis infection, rats were fed 10(9) viable L. casei bacteria 5 days per week. The T. spiralis-specific delayed-type hypersensitivity (DTH) response was significantly enhanced in both Wistar and Brown Norway rats given L. casei. In both rat strains fed L. casei, serum T. spiralis-specific immunoglobulin G2b (IgG2b) concentrations were also significantly increased. In the model, no significant effects of L. casei on larval counts or inflammatory reactions in the tongue musculature, body weights, or lymphoid organ weights were observed. Serum specific antibody responses, other than IgG2b, were not changed by feeding of L. casei. In contrast to L. casei, it was shown that orally administered Bifidobacterium breve or Bifidobacterium bifidum had no influence on the measured infection and immunity indices in the rat infection model. Since the rat DTH response is considered to be a manifestation of Th1 cell-mediated immunity and the IgG2b isotype has been associated with Th1 activity, it was concluded that Th1 cells could play an active role in the immunomodulatory effects of orally administered L. casei. Furthermore, our data do not indicate that the effect of oral supplementation with L. casei is dependent on the genetic background of the host. PMID- 11427424 TI - Identification and strain differentiation of Vibrio cholerae by using polyclonal antibodies against outer membrane proteins. AB - Cholera is caused only by O1 and O139 Vibrio cholerae strains. For diagnosis, 3 working days are needed for bacterial isolation from human feces and for biochemical characterization. Here we describe the purification of bacterial outer membrane proteins (OMP) from V. cholerae O1 Ogawa, O1 Inaba, and O139 strains, as well as the production of specific antisera and their use for fecal Vibrio antigen detection. Anti-OMP antisera showed very high reactivity and specificity by enzyme-linked immunosorbent assay (ELISA) and dot-ELISA. An inmunodiagnostic assay for V. cholerae detection was developed; this assay avoids preenrichment and costly equipment and can be used for epidemiological surveillance and clinical diagnosis of cases, considering that prompt and specific identification of bacteria is mandatory in cholera. PMID- 11427425 TI - Use of recombinant BP26 protein in serological diagnosis of Brucella melitensis infection in sheep. AB - Previously a Brucella protein named CP28, BP26, or Omp28 has been identified as an immunodominant antigen in infected cattle, sheep, goats, and humans. In the present study we evaluated antibody responses of infected and B. melitensis Rev.1 vaccinated sheep to the BP26 protein using purified recombinant BP26 protein produced in Escherichia coli in an indirect enzyme-linked immunosorbent assay (I ELISA). The specificity of the I-ELISA determined with sera from healthy sheep (n = 106) was 93%. The sensitivity of the I-ELISA assessed with sera from naturally infected and suspected sheep found positive in the current conventional diagnostic tests was as follows: 100% for bacteriologically and serologically positive sheep (n = 50), 88% for bacteriologically negative but serologically and delayed-type hypersensitivity-positive sheep (n = 50), and 84% for bacteriologically and serologically negative but delayed-type hypersensitivity positive sheep (n = 19). However, the absorbance values observed did not reach those observed in an I-ELISA using purified O-polysaccharide (O-PS) as an antigen. In sheep experimentally infected with B. melitensis H38 the antibody response to BP26 was delayed and much weaker than that to O-PS. Nevertheless, the BP26 protein appears to be a good diagnostic antigen to be used in confirmatory tests and for serological differentiation between infected and B. melitensis Rev.1-vaccinated sheep. Weak antibody responses to BP26 in some of the latter sheep suggest that a B. melitensis Rev.1 bp26 gene deletion mutant should be constructed to ensure this differentiation. PMID- 11427426 TI - Flow cytometric microsphere-based immunoassay: analysis of secreted cytokines in whole-blood samples from asthmatics. AB - The ability of flow cytometry to resolve multiple parameters was used in a microsphere-based flow cytometric assay for the simultaneous determination of several cytokines in a sample. The flow cytometer microsphere-based assay (FMBA) for cytokines consists of reagents and dedicated software, specifically designed for the quantitative determination of cytokines. We have made several improvements in the multiplex assay: (i) dedicated software specific for the quantitative multiplex assay that processes data automatically, (ii) a stored master calibration curve with a two-point recalibration to adjust the stored curve periodically, and (iii) an internal standard to normalize the detection step in each sample. Overall analytical performance, including sensitivity, reproducibility, and dynamic range, was investigated for interleukin-4 (IL-4), IL 6, IL-10, IL-12, gamma interferon (IFN-gamma), and tumor necrosis factor alpha. These assays were found to be reproducible and accurate, with a sensitivity in the picograms-per-milliliter range. Results obtained with FMBA correlate well with commercial enzyme-linked immunosorbent assay data (r > 0.98) for all cytokines assayed. This multiplex assay was applied to the determination of cytokine profiles in whole blood from atopic and nonatopic patients. Our results show that atopic subjects' blood produces more IL-4 (P = 0.003) and less IFN gamma (P = 0.04) than the blood of nonatopic subjects. However, atopic asthmatic subjects' blood produces significantly more IFN-gamma than that of atopic nonasthmatic subjects (P = 0.03). The results obtained indicate that the FMBA technology constitutes a powerful system for the quantitative, simultaneous determination of secreted cytokines in immune diseases. PMID- 11427427 TI - Antenatal screening for hepatitis B and antibodies to Toxoplasma gondii and rubella virus: evaluation of two commercial immunoassay systems. AB - A comparative evaluation of the Abbott AxSYM and DPC Immulite random-access analyzers was performed using 497 prospectively collected serum samples. These samples were sent to the laboratory for routine antenatal screening for hepatitis B surface antigen and immunoglobulin G (IgG) and IgM antibodies to Toxoplasma gondii and rubella virus. The overall agreement between the two assay systems ranged from 97.4 to 100%. After discrepancy analysis, the outcome in terms of sensitivity and specificity varied from 98.2 to 100% for all but one of the assays tested. The AxSYM rubella virus IgG assay tended to report protective or indeterminate antibody levels in 1% of the samples. This shortcoming might be overcome by raising the cutoff of the microparticle enzyme immunoassay system. PMID- 11427428 TI - Quantitative analyses of variations in the injury of endothelial cells elicited by 11 isolates of Rickettsia rickettsii. AB - Eleven isolates of spotted fever group rickettsiae from the blood of patients or ixodid ticks from North and South America were characterized. All isolates were identified as Rickettsia rickettsii using restriction fragment length polymorphism analysis of a 532-bp rOmpA gene fragment obtained by PCR. The ability of the R. rickettsii isolates to elicit cytopathic effects and parameters of oxidative injury were examined in cultured human EA.hy 926 endothelial cells. Cytopathic effects were determined by direct observation of infected cultures, by measuring the release of cytoplasmic lactate dehydrogenase (LDH), and by determination of intracellular pools of peroxide and reduced glutathione. Four biotypes of R. rickettsii were defined. Group I included two highly cytopathic isolates from Montana, Bitterroot and Sheila Smith, and three isolates from Maryland, North Carolina, and Brazil. These isolates rapidly damaged cells, released large amounts of cytoplasmic LDH, caused accumulation of intracellular peroxide, and depleted intracellular pools of reduced glutathione. Group II contained three isolates, two from Montana, Hlp#2 and Lost Horse Canyon, and an isolate from Colombia, which were similar to group I but caused either lower responses in LDH release or smaller changes in intracellular peroxide levels. The group III isolates, Sawtooth from Montana and 84JG from North Carolina, caused lower cellular injury by all measures. Group IV isolate Price T from Montana was the least cytopathic and caused minimal alterations of all parameters measured. Understanding the molecular basis for the varied cellular injury caused by different isolates of R. rickettsii may contribute to improved treatment of Rocky Mountain spotted fever and to the rapid identification of those isolates which are more likely to cause fulminant disease. PMID- 11427429 TI - AhpC, AhpD, and a secreted 14-kilodalton antigen from Mycobacterium avium subsp. paratuberculosis distinguish between paratuberculosis and bovine tuberculosis in an enzyme-linked immunosorbent assay. AB - Sera from cattle naturally infected with Mycobacterium avium subsp. paratuberculosis (n = 56) and naturally (n = 4) and experimentally (n = 8) infected with Mycobacterium bovis were tested for the presence of antibodies against paratuberculosis antigens. An enzyme-linked immunosorbent assay (ELISA) was established based on absorption of M. avium subsp. paratuberculosis antigens on a hyperimmune antiserum against M. avium subsp. avium proteins in order to remove cross-reacting antigens. This absorbed-antigen ELISA recognized 66% of animals with paratuberculosis (37 of 56), while none of the animals with naturally occurring bovine tuberculosis (TB) had detectable antibodies. However, the animals with experimental bovine TB also responded in this ELISA. Similar results were found in a commercial ELISA, showing that neither of these tests was able to distinguish between paratuberculosis and bovine TB. The sera were further tested for antibody activities against purified AhpC and AhpD, which are proteins constitutively expressed by M. avium subsp. paratuberculosis, and against a secreted 14-kDa protein present in culture filtrates from the M. avium complex. Elevated antibody levels to AhpC, AhpD, and the 14-kDa antigen were found in 27% (13 of 48), 15% (7 of 48), and 27% (13 of 48), respectively, of the cattle with paratuberculosis. Together these ELISAs were positive with 35% (17 of 48) of the animals. None of the animals with bovine TB had detectable antibodies against any of the purified proteins despite their high levels of cross-reacting antibodies. These results show that purified specific antigens are needed to differentiate between paratuberculosis and bovine TB in ELISA. PMID- 11427430 TI - Antibody maturation in Trypanosoma cruzi-infected rats. AB - The study of antibody avidity changes during infection has improved the understanding of the pathologic processes involved in several infectious diseases. In some infections, like toxoplasmosis, this information is being used for diagnostic purposes. Results of the evolution of antibody avidity for different specific antigens in Trypanosome cruzi-infected rats are presented. A Western blotting technique, combined with avidity analysis to identify antigens that elicit high-avidity antibodies, is suggested. In this system, antibodies showed high avidity values only during the chronic phase of infection and only in relation to antibodies against 21-, 33-, 41-, 42-, 56-, 58-, 66-, and 72-kDa antigens. Finally, a 97-kDa T. cruzi antigen, which was recognized by high avidity antibodies and occurred in noninfected rats, was identified. These results allow us to evaluate the different antigens in chagasic infection. Our results show that with the correct choice of antigen it is possible to detect differences in maturation of antibodies and to discriminate, in an experimental model, between recent (acute) and chronic infections. PMID- 11427431 TI - Enhancement of splenic-macrophage Fcgamma receptor expression by treatment with estrogens. AB - Splenic-macrophage Fcgamma receptors (FcgammaRs) participate in the pathophysiologies of immune-complex diseases and in host defense against infection. Modulation of macrophage FcgammaR expression is an immuno-therapeutic target. Glucocorticoids, sex steroids, and dopaminergic drugs modulate macrophage FcgammaR expression. Previous data indicate that estradiol increases macrophage FcgammaR expression. Nevertheless, the effects of clinically used estrogens upon macrophage FcgammaR expression are unknown. We assessed the effects of treatment with commonly used estrogens on the expression of macrophage FcgammaRs using a guinea pig experimental model. Six estrogens have been studied: ethynylestradiol (Et), mestranol (M), chlortianisene (Ct), promestriene, 17-epiestriol, and 17beta estradiol. Following in vivo treatment of guinea pigs, we determined the clearance of immunoglobulin G (IgG)-sensitized erythrocytes in vivo, the binding of IgG-sensitized erythrocytes by isolated splenic macrophages, and splenic macrophage FcgammaR cell surface expression. Estrogens enhance the clearance of IgG-sensitized erythrocytes by increasing splenic-macrophage FcgammaR expression. Et, M, and Ct were more effective than the other estrogens. Flow cytometry and fluorescence microscopy with monoclonal antibodies demonstrated that estrogens increase the cell surface expression of FcgammaR1 and -2 more than that of FcgammaR2. These data indicate that treatment with commonly used estrogens enhances the clearance of IgG-sensitized cells by improving splenic-macrophage FcgammaR expression. PMID- 11427432 TI - In the absence of endogenous gamma interferon, mice acutely infected with Neospora caninum succumb to a lethal immune response characterized by inactivation of peritoneal macrophages. AB - Following infection with Neospora caninum, BALB/c mice were shown to be resistant to an acute infection but developed a latent chronic infection. However, BALB/c background gamma interferon (IFN-gamma)-deficient mice were sensitive to the acute infection. Since the immune response in IFN-gamma-deficient mice is scantly known, we examined the function of macrophages, major histocompatibility complex (MHC) class II expression, T-cell responses, and serum cytokine levels in the mice. All IFN-gamma-deficient mice died within 9 days of infection with N. caninum, whereas those treated with exogenous IFN-gamma lived longer. Although N. caninum invaded various organs in both types of mice at the early stage of infection, the parasite was not detected in the brains of resistant hosts until 21 days postinfection (dpi). Peritoneal macrophages from IFN-gamma-deficient mice were activated by exogenous IFN-gamma associated with inhibition of parasite growth and nitric oxide production as were those from BALB/c mice. IFN-gamma deficient mice failed to increase MHC class II expression on macrophages. Moreover, BALB/c mice induced T-cell proliferation while IFN-gamma-deficient mice did not. However, in vivo treatment with exogenous IFN-gamma induced up-regulated MHC class II expression in IFN-gamma-deficient mice. BALB/c mice treated with an antibody to CD4 showed an increase in morbidity and mortality after parasite infection. In serum, significant levels of IFN-gamma and interleukin-4 (IL-4) were detected in resistant hosts, whereas IL-10 was detected in IFN-gamma deficient mice. The levels of IL-12 in IFN-gamma-deficient mice were higher than those in BALB/c mice at 7 dpi. The present study indicates that early IFN-gamma production has a crucial role in the activation of peritoneal macrophages for the induction of protective immune responses against N. caninum. PMID- 11427433 TI - Viral load in breast milk correlates with transmission of human cytomegalovirus to preterm neonates, but lactoferrin concentrations do not. AB - In vitro, lactoferrin (LF) strongly inhibits human cytomegalovirus (HCMV), which led us to hypothesize that in vivo HCMV might also be inhibited in secretions with high LF concentrations. In breast milk, high viral loads observed as high viral DNA titers tended to coincide with higher LF levels. However, the LF levels did not correlate to virus transmission to preterm infants. The viral load in the transmitting group was highest compared to the nontransmitting group. We conclude that viral load in breast milk is an important factor for transmission of the virus. PMID- 11427434 TI - Neutralizing antibodies against autologous human immunodeficiency virus Type 1 isolates in patients with increasing CD4 cell counts despite incomplete virus suppression during antiretroviral treatment. AB - Antiretroviral-treated human immunodeficiency virus (HIV) type 1-seropositive individuals can remain clinically stable for a long period of time with an increasing CD4 cell count irrespective of incomplete viral suppression. We evaluated the role of neutralizing antibody (NtAb) activity in the etiopathogenesis of this viro-immunological disconnection (defined as an increasing CD4(+)-cell count despite a persistent, detectable viral load during antiretroviral therapy) in 33 patients failing therapy with two analogue nucleoside reverse transcriptase inhibitors. An HIV NtAb titer of >/=1:25 was detected in specimens from 16 out of 33 (48%) patients. A significant correlation was found between NtAb titers and CD4(+)-cell counts (P = 0.001; r = 0.546) but not with HIV RNA levels in plasma. Five patients with a viro-immunological disconnection had an NtAb titer of >1:125, statistically higher than the NtAb titers for the remaining 28 patients with both virologic and immunologic failure (P < 0.0001). The HIV-specific humoral immune response could play a role during antiretroviral treatment to improve immunological function despite an incomplete suppression of viral load. PMID- 11427435 TI - Generation and serological characterization of murine monoclonal antibodies against O antigens from Acinetobacter reference strains. AB - O-antigen-specific monoclonal antibodies were generated against Acinetobacter strains from international type culture collections and characterized by enzyme immunoassay and Western and colony blotting. The antibodies aid in the further completion of an O-serotyping scheme for Acinetobacter and, due to their high specificity, are especially useful to all working with these strains. PMID- 11427436 TI - Real-time PCR as a new tool for quantifying Leishmania infantum in liver in infected mice. AB - The parasitic loads of mouse livers experimentally infected with Leishmania infantum were determined using a double real-time quantitative PCR test targeted to the parasite DNA polymerase gene and to the mouse brain-derived neutrophic factor gene. The Leishmania DNA copy number was normalized to the number of mouse gene copies in order to quantify the former independently of liver weight. The correlation coefficient with the microtitration method was 0.66. This PCR assay can be considered for experimental pharmaceutical studies. PMID- 11427437 TI - groEL encodes a highly antigenic protein in Burkholderia pseudomallei. AB - No recombinant protein is available for serodiagnosis of melioidosis. In this study, we report the cloning of the groEL gene, which encodes an immunogenic protein of Burkholderia pseudomallei. Bidirectional DNA sequencing of groEL revealed that the gene contained a single open reading frame encoding 546 amino acid residues with a predicted molecular mass of 57.1 kDa. Basic Local Alignment Search Tool analysis showed that the putative protein encoded by groEL is homologous to the chaperonins encoded by the groEL genes of other bacteria. It has 98% amino acid identity with the GroEL of Burkholderia cepacia, 98% amino acid identity with the GroEL of Burkholderia vietnamiensis, and 82% amino acid identity with the GroEL of Bordetella pertussis. Furthermore, it was observed that patients with melioidosis develop a strong antibody response against GroEL, suggesting that the recombinant protein and its monoclonal antibody may be useful for serodiagnosis in patients with melioidosis and that the protein may represent a good cell surface target for host humoral immunity. Further studies in these directions would be warranted. PMID- 11427438 TI - Cytopathic changes in rat microglial cells induced by pathogenic Acanthamoeba culbertsoni: morphology and cytokine release. AB - To determine whether pathogenic Acanthamoeba culbertsoni trophozoites and lysate can induce cytopathic changes in primary-culture microglial cells, morphological changes were observed by transmission electron microscopy (TEM). In addition, the secretion of two kinds of cytokines, tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta), from microglial cells was observed. Trophozoites of pathogenic A. culbertsoni made contact with microglial cells and produced digipodia. TEM revealed that microglial cells cocultured with amoebic trophozoites underwent a necrotic process, accompanied by lysis of the cell membrane. TEM of microglial cells cocultured with amoebic lysate showed that the membranes of the small cytoplasmic vacuoles as well as the cell membrane were lysed. The amounts of TNF-alpha secreted from microglial cells cocultured with A. culbertsoni trophozoites or lysate increased at 6 h of incubation. The amounts of IL-1beta secreted from microglial cells cocultured with A. culbertsoni trophozoites at 6 h of incubation was similar to those secreted from the control group, but the amounts decreased during cultivation with A. culbertsoni lysate. These results suggest that pathogenic A. culbertsoni induces the cytopathic effects in primary-culture rat microglial cells, with the effects characterized by necrosis of microglial cells and changes in levels of secretion of TNF-alpha and IL-1beta from microglial cells. PMID- 11427439 TI - Syphilis fast latex agglutination test, a rapid confirmatory test. AB - Using 255 serum samples with various reactivities, we evaluated the Syphilis Fast latex agglutination test (Syphilis Fast) against the Treponema pallidum particle agglutination test (TP-PA) for confirming a diagnosis of syphilis. We found 98.8% agreement between the Syphilis Fast and the TP-PA. The Syphilis Fast, however, had a couple of advantages over the TP-PA: the test takes only 8 min to perform and produces results that are easy to read. It appears to be a good confirmatory test for syphilis, especially for point-of-care clinics such as prenatal or sexually transmitted disease clinics. PMID- 11427440 TI - Heartwater (Cowdria ruminantium infection) as a cause of postrestocking mortality of goats in Mozambique. AB - A serological survey in Mozambique to detect antibodies to Cowdria ruminantium, the etiologic agent of heartwater, revealed a seroprevalence of 8.1% (n = 332) for goats in the northern province of Tete and of 65.6% (n = 326) for goats in the southern provinces. Translocation of 10 serologically negative goats from Tete to farms in the south resulted in two clinical cases of heartwater that were fatal. In addition, four goats seroconverted within the study period of 5 weeks. One goat showed no symptoms. Two goats died of other causes, whereas the remaining goat went missing after 1 week. Experimental needle infections of goats and sheep were conducted to confirm results and to isolate different strains of C. ruminantium. These data indicate that translocation of goats from the north to the south of Mozambique bears a high risk of C. ruminantium infection, which can cause fatal disease. PMID- 11427441 TI - Production of Bartonella genus-specific monoclonal antibodies. AB - Monoclonal antibodies (MAbs) which react with heat-resistant proteins with molecular masses of 32 to 33 kDa of 14 different Bartonella species were produced. These antibodies did not react with antigens of 26 diverse bacterial strains by microimmunofluorescence assay except MAb B3D4, which reacted with Chlamydia psittaci and Chlamydia trachomatis at low titers. The identification of a common Bartonella antigenic protein will make it possible to later produce a diagnostic antigen by cloning and expressing it in Escherichia coli. Moreover, these MAbs allow all Bartonella species to be identified to the genus level. PMID- 11427442 TI - Spinal cord involvement in uncomplicated herpes zoster. AB - We prospectively evaluated herpes zoster patients during the acute phase of the disease for central nervous system involvement. Of 24 patients with spinal zoster, 13 (54%) had spinal cord abnormality, which was asymptomatic in 12 of the 13. Age but not lack of acyclovir treatment was associated with such involvement. In all but 2, neurological involvement resolved within 6 months. Although the mechanism responsible for the neurological abnormalities is unknown, findings may support the hypothesis that zoster is associated with spread of viral infection into the spinal cord and therefore support the possibility that zoster is due to active viral replication in the ganglion. PMID- 11427443 TI - The metabolic autopsy comes of age. PMID- 11427444 TI - Hereditary hemochromatosis since discovery of the HFE gene. AB - BACKGROUND: Hereditary hemochromatosis is an inherited disorder of iron metabolism that is characterized by excessive iron deposition in major organs of the body. Chronic increased iron absorption leads to multiorgan dysfunction. Since the discovery of the gene responsible for the majority of cases, research has progressed rapidly to identify the gene product, the effects of mutations, and the implications for different populations. The protein product of the HFE gene is a transmembrane glycoprotein, termed HFE, that modulates iron uptake. Mutations in the HFE protein compromise its function and produce disease symptoms. Two mutations, C282Y and H63D, have been linked to the majority of disease cases. APPROACH: We reviewed the recent literature for the molecular basis of hereditary hemochromatosis. Genotypic information was combined with biochemical and clinical phenotypic information to achieve a better understanding of the disease mechanism. CONTENT: This review provides a comprehensive discussion of known mutations in the HFE gene and their phenotypic expression. Diagnostic criteria using molecular genetic techniques in conjunction with traditional biochemical tests are provided. Current methods and limitations of molecular testing are examined in detail. A strategy for population screening and an algorithm for diagnosis that incorporates molecular testing are presented. Treatment by therapeutic phlebotomy and the use of blood obtained from hemochromatosis patients are discussed. SUMMARY: Although the disease mechanism has not been completely elucidated, phenotypic and penetrance data are becoming available. Controversy still exists concerning the role of genetic testing in diagnosis and population screening. PMID- 11427445 TI - Glycohemoglobin: a primary predictor of the development or reversal of complications of diabetes mellitus. AB - BACKGROUND: Diabetes mellitus is a major health problem worldwide with long-term micro- and macrovascular complications responsible for a majority of its morbidity and mortality. The development and progression of these complications relate strongly to glycemic control. METHODS: We reviewed the literature extensively for studies that relate glycemic control to the development and progression of diabetic complications. We discuss the problems of standardizing glycohemoglobin measurements for monitoring diabetic therapy and also consider recently developed electrospray ionization mass spectrometry methods that have been considered as candidate reference methods for estimation of glycohemoglobin. RESULTS: Several clinical trials and studies have clearly shown that improved glycemic control is strongly associated with decreased development and/or progression of complications in both type 1 and type 2 diabetes mellitus. Irrespective of the methods used for estimating glycohemoglobin, these results underline the importance of glycohemoglobin for guiding therapy of diabetes mellitus. Recently developed candidate reference methods promise to yield greatly improved standardization for the measurement of glycohemoglobin. CONCLUSIONS: Glycohemoglobin measurement remains the optimal indicator of glycemic control in diabetic patients, but translation of findings from clinical trials to clinical practice worldwide demands consistent values across all assays. To ensure that the important prognostic information still applies to all diabetic patients with the application of the reference method(s), the hemoglobin A(1c) values reported in the major clinical trials will have to be translated into statistically and computationally compatible values based on the new reference system(s). PMID- 11427446 TI - Electrospray tandem mass spectrometry for analysis of acylcarnitines in dried postmortem blood specimens collected at autopsy from infants with unexplained cause of death. AB - BACKGROUND: Deaths from inherited metabolic disorders may remain undiagnosed after postmortem examination and may be classified as sudden infant death syndrome. Tandem mass spectrometry (MS/MS) may reveal disorders of fatty acid oxidation in deaths of previously unknown cause. METHODS: We obtained filter paper blood from 7058 infants from United States and Canadian Medical Examiners. Acylcarnitine and amino acid profiles were obtained by MS/MS. Specialized interpretation was used to evaluate profiles for disorders of fatty acid, organic acid, and amino acid metabolism. The analyses of postmortem blood specimens were compared with the analyses of bile specimens, newborn blood specimens, and specimens obtained from older infants at risk for metabolic disorders. RESULTS: Results on 66 specimens suggested diagnoses of metabolic disorders. The most frequently detected disorders were medium-chain and very-long-chain acyl-CoA dehydrogenase deficiencies (23 and 9 cases, respectively), glutaric acidemia type I and II deficiencies (3 and 8 cases, respectively), carnitine palmitoyl transferase type II/translocase deficiencies (6 cases), severe carnitine deficiency (4 cases), isovaleric acidemia/2-methylbutyryl-CoA dehydrogenase deficiencies (4 cases), and long-chain hydroxyacyl-CoA dehydrogenase/trifunctional protein deficiencies (4 cases). CONCLUSIONS: Postmortem metabolic screening can explain deaths in infants and children and provide estimates of the number of infant deaths attributable to inborn errors of metabolism. MS/MS is cost-effective for analysis of postmortem specimens and should be considered for routine use by Medical Examiners and pathologists in unexpected/unknown infant and child death. PMID- 11427447 TI - In-tube DNA methylation profiling by fluorescence melting curve analysis. AB - BACKGROUND: Most PCR assays for detection of 5-methylcytosine in genomic DNA entail a two-step procedure, comprising initial PCR amplification and subsequent product analysis in separate operations that usually require manual transfer. These methods generally provide information about methylation of only a few CpG dinucleotides within the target sequence. METHODS: An in-tube methylation assay is described that integrates amplification of bisulfite-treated DNA and melting analysis by using a thermal cycler coupled to a fluorometer (LightCycler). DNA melting curves were acquired by measuring the fluorescence of a double-stranded DNA-binding dye (SYBR Green I) during a linear temperature transition. RESULTS: Analysis of a region comprising 11 CpG sites at the SNRPN promoter CpG island showed that the melting temperature (T(m)) differed by approximately 3 degrees C between unmethylated and fully methylated alleles. This assay could easily distinguish patients with Prader-Willi syndrome or Angelman syndrome from individuals without these conditions. Melting curve analysis also allowed resolution of methylation "mosaicism" at the p15(Ink4b) promoter in bone marrow samples from patients with acute myeloid leukemia (AML). AML samples representing pools of heterogeneously methylated p15(Ink4b) alleles showed broadened melting peaks with overall T(m)s between those of the unmethylated and fully methylated alleles. CONCLUSIONS: Integration of PCR and fluorescence melting analysis may be useful for simple and cost-effective detection of aberrant methylation patterns. PMID- 11427448 TI - Accumulation of free 3-hydroxy fatty acids in the culture media of fibroblasts from patients deficient in long-chain l-3-hydroxyacyl-CoA dehydrogenase: a useful diagnostic aid. AB - BACKGROUND: The diagnosis of long-chain L-3-hydroxy-acyl-coenzyme A dehydrogenase (LCHAD) deficiency frequently requires the study of cultured fibroblasts. We developed such a test that does not require disruption and loss of the cells. METHODS: We measured free 3-hydroxy fatty acids (3-OHFAs) in media of skin fibroblasts cultures from 11 patients with a genetic deficiency of LCHAD and the associated disorder of mitochondrial trifunctional protein (MTFP). Fibroblasts were cultured for 24 h with 100 micromol/L nonisotopic palmitate added. 3-OHFAs were measured by selected-ion monitoring, stable-isotope dilution gas chromatography-mass spectrometry with [(13)C]-labeled internal standards. RESULTS: 3-OH-hexadecanoic and 3-OH-tetradecanoic FAs were increased 14- and 11 fold, respectively, in all patients with LCHAD or MTFP deficiency when compared with control fibroblast cell lines after overnight incubation with palmitate. 3 OH-dodecanoic FA demonstrated a modest, fivefold increase in LCHAD-deficient cells. The concentrations of all 3-OHFAs were similar whether or not the medium samples were hydrolyzed to release conjugated species such as acylcarnitines, suggesting that 3-OHFAs accumulate in the media as free FAs. CONCLUSIONS: Measurement of 3-OHFA excretion from LCHAD- or MTFP-deficient cell lines can be used as a diagnostic tool. Free FAs are the predominant form of these abnormal metabolic intermediates in culture media. PMID- 11427449 TI - Capillary electrophoresis-based heteroduplex analysis with a universal heteroduplex generator for detection of point mutations associated with rifampin resistance in tuberculosis. AB - BACKGROUND: Slab gel heteroduplex analysis (HDA), a popular scanning method for genetic mutations, uses DNA fragments typically generated by PCR to create homo- and heteroduplex molecules with conformational differences and sequence-dependent electrophoretic profiles. Use of a universal heteroduplex generator (UHG) enhances the subtle variations caused by single-base substitutions. METHODS: The HDA-UHG slab gel format was modified for an efficient capillary-based method. The effect of staining dyes TOPRO5 and YOPRO1 on the analysis of heteroduplexes was studied, as well as ultraviolet absorbance and laser-induced fluorescence (LIF) detection methods. In addition, the entangled polymers hydroxyethyl cellulose, methyl cellulose, and linear polyacrylamide were evaluated as separation matrices. RESULTS: This assay was able to detect the presence of Mycobacterium tuberculosis and its rifampin susceptibility directly from clinical specimens in dramatically reduced analysis time (30 min vs 2.5 h). Optimized conditions included 0.3% methyl cellulose as the separation matrix, on-line staining using 1 micromol/L YOPRO1, and LIF detection for quantitative and reproducible analysis of single-base substitutions in the rifampin resistance-determining region of rpoB that give rise to the rifampin-resistant phenotype of M. tuberculosis. We generated 95% confidence limits using the wild-type sequence and used these limits to determine rifampin susceptibility in samples. CONCLUSIONS: Capillary electrophoresis, combined with the HDA-UHG technique, may be of value for rapid and efficient clinical diagnosis of rifampin-resistant tuberculosis strains. PMID- 11427450 TI - Discontinuation of the bleeding time test without detectable adverse clinical impact. AB - BACKGROUND: The bleeding time (BT) test predicts a higher bleeding complication rate in populations at risk for inherited or acquired platelet dysfunction, but it is of limited assistance in evaluating individual patients. There are no reports of clinical outcomes after discontinuation of the BT test. METHODS: Interviews with a subset of the physicians who had ordered the BT test before discontinuation of the test were conducted. The total number of platelet aggregation tests, the mean number of monthly, unmodified platelet units transfused, the incidence of kidney biopsy complications, and the number of doses of 1-deamino-8-D-arginine vasopressin (DDAVP) administered 5 months before and after discontinuation of the BT test were compared. We recorded the rates of bleeding complications in the Major Surgery Risk Pool during the 12 months before and the 5 months after the discontinuation of the BT test. RESULTS: Clinicians reported they did not significantly change their preprocedural work-ups, postpone an invasive procedure, experience an increase in bleeding complications, or increase their use of blood products after discontinuation of the BT test. Platelet-aggregation tests (n = 9, before and after), platelet transfusions (P = 0.958), and DDAVP administration (before = 24; after = 10) did not increase after discontinuation of the BT test. The rate of postprocedural bleeding complications did not increase significantly in either Major Surgery Risk Pool cases (<3final sigma deviation from the mean rate) or in patients undergoing renal biopsies (P = 0.225 for decrease in hematocrit; P = 1.000 for the percentage of patients transfused) after discontinuation of the BT test. CONCLUSIONS: Our study failed to identify a clinically significant, negative impact of discontinuing the BT test. PMID- 11427451 TI - Interpretation of hemoglobin A(1c) (HbA(1c)) values among diabetic patients: implications for quality specifications for HbA(1c). AB - BACKGROUND: Few studies have examined patients' views, knowledge, and understanding of glycohemoglobin A(1c) (HbA(1c)) testing. We explored such issues in patients with type 1 diabetes and used their statements to estimate analytical quality specifications for HbA(1c) testing. METHODS: We recruited 201 patients from a hospital outpatient clinic. A questionnaire was used to collect information on diabetes characteristics, perceived knowledge of HbA(1c), last HbA(1c) value, HbA(1c) target value, and thresholds for action. Patients were asked to indicate the magnitude of change in HbA(1c) from 9.4% that they would consider to be a true (real) change; from their responses, we calculated patient derived quality specifications for HbA(1c). RESULTS: Fifty-eight percent of the patients felt they had "high" knowledge about HbA(1c), and >80% of responders knew their last HbA(1c) value, their target HbA(1c), and the threshold value of HbA(1c) for treatment intensification. The mean acceptable HbA(1c) value was 7.5%. Patients with lower values on their most recent tests reported lower target values for HbA(1c) and lower values for the upper HbA(1c) threshold for treatment intensification. An analytical CV (CV(a)) of 3.1% would be satisfactory for 75% of patients when HbA(1c) is increasing (80% confidence), and a CV(a) of 3.2% would be satisfactory for 75% when HbA(1c) is decreasing (95% confidence). CONCLUSIONS: Type 1 patients' perceived knowledge about HbA(1c) testing is high. They are well informed about their own personal results and about target values and the upper HbA(1c) threshold for action. The patient-derived analytical quality specification for imprecision (CV) is 3.1%. PMID- 11427452 TI - Development of a dual monoclonal antibody immunoassay for total human kallikrein 2. AB - BACKGROUND: Human kallikrein 2 (hK2) shares 80% sequence identity with prostate specific antigen (PSA). Because both hK2 and hK2-alpha(1)-antichymotrypsin (hK2 ACT) complexes have been identified in patient sera, we devised an immunoassay for total hK2 [(thK2); hK2 and hK2-ACT] and evaluated it in healthy subjects and patients with prostate disease. METHODS: We developed monoclonal antibodies (mAbs) with high specificity for hK2 and hK2-ACT and minimal cross-reactivity to PSA. Using these mAbs, a sandwich assay was developed and its specificity for forms of hK2 was assessed. Serum samples (n = 1035) from healthy volunteers, patients with increased PSA, and men who had undergone radical prostatectomy were assayed for thK2. We also measured thK2 in samples before and after storage under common laboratory conditions. RESULTS: The minimum detectable concentration in the thK2 assay was 0.008 microg/L, and PSA cross-reactivity was <0.001%. The assay detected prohK2 and three different hK2-serum protease complexes. The median serum concentration of thK2 in control samples (0.013 microg/L) was significantly lower than the median in samples from patients with increased PSA concentrations (0.085 microg/L). Immunoreactive hK2 changed little in samples stored for up to 1 month at -70 degrees C. CONCLUSIONS: The thK2 assay recognizes all forms of hK2 that have been found in bodily fluids to date. PMID- 11427453 TI - Interference of transferrin isoform types with carbohydrate-deficient transferrin quantification in the identification of alcohol abuse. AB - BACKGROUND: Isoforms of transferrin interfere with measurement of carbohydrate deficient transferrin (CDT) as a marker of heavy alcohol consumption. We evaluated the rate of inaccurate CDT results by immunoassays. METHODS: We studied 2360 consecutive sera (1614 individuals) submitted for CDT assay without clinical information as well as samples from 1 patient with a congenital disorder of glycosylation (CDG Ia) and from 6 healthy carriers of CDG Ia. The CDTect, %CDT TIA, and new %CDT immunoassays were compared with HPLC (%CDT-HPLC). Transferrin isoform pattern were evaluated by isoelectric focusing (IEF). RESULTS: Transferrin BC and CD heterozygotes were found at frequencies of approximately 0.7% and approximately 0.2%, respectively. Another transferrin C subtype, where di- and trisialotransferrin partly coeluted (tentatively identified as C2C3), was observed in approximately 0.6%. Compared with the %CDT-HPLC method, the immunoassays often produced low results for transferrin BC and high results for transferrin CD and "C2C3". A very high trisialotransferrin value (frequency approximately 1%) often produced high CDT immunoassay results. In four of six healthy carriers of CDG Ia, a- and disialotransferrin were highly increased and the HPLC and IEF isoform patterns were indistinguishable from those in alcohol abuse. CONCLUSIONS: Rare transferrin isoform types and abnormal amounts of trisialotransferrin (total frequency approximately 2-3%) may cause incorrect determination of CDT with immunoassays. The observed variants were readily identified by HPLC and IEF, which can be recommended for verification of CDT immunoassay results in doubtful cases. In healthy carriers of CDG Ia, CDT is high by all assays. PMID- 11427454 TI - Enhanced susceptibility to oxidation and diminished vitamin E content of LDL from patients with stable coronary artery disease. AB - BACKGROUND: Convincing evidence points to oxidative modification of LDL as an important trigger in a complex chain of events leading to atherosclerosis. We investigated the occurrence of enhanced susceptibility of LDL to oxidation and decreased vitamin E concentration in LDL as additional risk factors promoting atherosclerosis among patients with established coronary artery disease (CAD). METHODS: We examined 132 patients with angiographically confirmed CAD and compared them with 111 healthy control individuals. We measured conjugated diene production to assess susceptibility of LDL to copper-mediated oxidation. Vitamin E content of LDL was measured by HPLC. RESULTS: The mean lag time of LDL oxidation and LDL alpha-tocopherol/LDL-cholesterol ratio were lower in the patients with CAD (55 +/- 14 min and 2.4 +/- 1.0 mmol/mmol) than in the controls (63 +/- 13 min and 2.9 +/- 1.1 mmol/mmol; P <0.0001 and <0.001, respectively). Multiple stepwise regression analysis demonstrated the lag time (odds ratio, 1.96; 95% confidence interval, 1.34-2.87; P <0.0001) and concentration of vitamin E in LDL (odds ratio, 1.65; 95% confidence interval, 1.16-2.33; P <0.005) as independent determinants of CAD. Significant inverse Spearman rank correlations were found between lag time (r = -0.285; P <0.001) or concentration of vitamin E in LDL (r = -0.197; P <0.002) and severity of CAD. Lag times were not significantly correlated with serum C-reactive protein or ferritin. CONCLUSIONS: Our data suggest that a short LDL oxidation lag time and a low concentration of vitamin E in LDL might be independent coronary risk factors for stable CAD in Iranian people. PMID- 11427455 TI - Pharmacokinetic basis for the efficient and safe use of low-dose mycophenolate mofetil in combination with tacrolimus in kidney transplantation. AB - BACKGROUND: Mycophenolate mofetil (MMF) is an effective posttransplantation immunosuppressive agent used in combination with cyclosporin A (CsA) or tacrolimus (Tc). An increase in plasma mycophenolic acid (MPA) has been shown in patients receiving Tc-MMF combination therapy compared with CsA-MMF combination therapy at the same dose of MMF. The aim of this prospective study was to assess the pharmacokinetic/pharmacodynamic (PK/PD) relationship for MPA in kidney transplant patients receiving low-dose MMF (500 mg twice a day) in combination with Tc. METHODS: Adult kidney transplant recipients (n = 51) were included. MPA PK profiles (blood sampling at 0, 0.5, 1, 2, 4, 6, and 12 h after MMF oral dose) were obtained within the first 2 weeks after transplantation, 3 months after grafting, and at every adverse clinical event [side effect or acute rejection (AR)]. All patients received Tc, MMF (500 mg twice a day), and steroids. RESULTS: Thirty patients (59%) had uneventful outcomes, and 21 patients had 33 episodes of MPA-related side effects; only 3 patients had AR. A total of 78 MPA-PK profiles were obtained. The following PK parameters were increased in the side-effects group compared with the non-side effects group: mean MPA c(min), 2.63 +/- 1.58 vs 1.75 +/- 0.82 mg/L (P = 0.016); mean c(30), 10.47 +/- 6.27 vs 7.66 +/- 8.95 mg/L (P = 0.009); mean c(60), 9.67 +/- 5.42 vs 5.83 +/- 2.6 mg/L (P = 0.0002); mean area under the MPA time-concentration curve from 0 to 12 h [MPA-AUC((0-12))], 48.38 +/- 18.5 vs 36.04 +/- 10.82 mg. h/L (P = 0.0006); mean dose-normalized MPA AUC, 0.16 +/- 0.05 vs 0.12 +/- 0.04 (mg. h/L)/(mg/m(2)) (P = 0.0015). For the three AR patients, MPA concentrations obtained at the time of AR revealed MPA c(min) values of 1.86, 1.76, and 3.83 mg/L, respectively, and MPA-AUC((0-12)) values of 37.7, 24.9, and 104.9 mg. h/L. The threshold of toxicity was 3 mg/L (sensitivity, 38.7%; specificity, 91.5%) for c(min), 8.09 mg/L for maximum MPA concentration during the first hour (sensitivity, 77.8%; specificity, 67.4%), and 37.6 mg. h/L for MPA-AUC((0-12)) (sensitivity, 83.3%; specificity, 59.6%). CONCLUSIONS: These results demonstrate the relationship between plasma MPA concentrations and toxicity. High c(min), c(30), and c(60) values as well as AUC((0-12)) are associated with increased risk for side effects. These values may have an importance in a routine monitoring program. PMID- 11427456 TI - Immunochemical assay of hemoglobin with N(epsilon)-(carboxymethyl)lysine at lysine 66 of the beta chain. AB - BACKGROUND: N(epsilon)-(Carboxymethyl)lysine (CML), a well-characterized and major advanced glycation end product structure, is produced via a Maillard reaction by nonenzymatic glycation and/or oxidation. Although few of the carboxymethylation sites of lysine residues on proteins have been identified, it is known that the possible lysine glycation site in hemoglobin (Hb) is Lys-66 on the beta chain. We aimed to develop an assay for the Hb with a CML (CML-Hb) site specific to Lys-66 on the Hb beta chain and to determine whether the lysine residue at that site is carboxymethylated. METHODS: Ala-His-Gly-Lys-Lys(CM)-Val Leu-Gly-Ala-Phe-Ser-Cys, the peptide derived from the beta chain of human Hb, was synthesized as an immunogen, and a monoclonal antibody against the peptide was prepared. A latex immunoassay method was established using the antibody on an automatic analyzer. In this study, 20 samples from healthy subjects and 80 samples from nondiabetic patients undergoing hemodialysis (HD) were analyzed. RESULTS: The latex immunoassay method using the antibody correlated significantly with the ELISA method using the antibody (r = 0.95; P <0.001). Between healthy subjects (n = 20) and nondiabetic HD patients (n = 80), a significant difference was seen in circulating CML-Hb (525 +/- 76 vs 778 +/- 137 pmol CML/mg of Hb; P <0.0001). CONCLUSION: The latex method for the CML-Hb site specific to Lys-66 on the beta chain can measure large numbers of samples on an automatic analyzer. PMID- 11427457 TI - Quantitative determination of estradiol fatty acid esters in human pregnancy serum and ovarian follicular fluid. AB - BACKGROUND: Lipophilic estradiol derivatives carried by lipoprotein particles in blood may mediate antioxidant or endocrine effects. We developed a new quantitative method to determine the concentration of circulating lipophilic estradiol fatty acid esters in human early- and late-pregnancy serum and in ovarian follicular fluid. METHODS: After extraction from serum or follicular fluid, estradiol fatty acid esters were separated from nonesterified estradiol by Sephadex LH-20 column chromatography. The estradiol ester fraction was hydrolyzed by saponification and further purified by several chromatographic steps. The hydrolyzed estradiol esters were measured by time-resolved fluoroimmunoassay. RESULTS: The average estradiol fatty acid ester concentration in serum increased 10-fold during pregnancy, from 40.4 pmol/L (expressed as pmol/L estradiol; range, 25.0-64.2 pmol/L) in early pregnancy (n = 8) to 404 pmol/L (196-731 pmol/L) in late pregnancy (n = 10). The ratio of estradiol ester to nonesterified estradiol remained relatively constant during pregnancy, at 0.4-0.6%. In 10 follicular fluid samples, the mean estradiol ester concentration was 106 nmol/L (56.9-262 nmol/L). Compared with serum, a greater proportion of estradiol in follicular fluid (3.0-10%) was in the esterified form. CONCLUSION: The new method provides a means to measure circulating estradiol fatty acid ester concentrations in human pregnancy serum. PMID- 11427458 TI - Cystatin C is an independent predictor of fasting and post-methionine load total homocysteine concentrations among stable renal transplant recipients. AB - BACKGROUND: An increased prevalence of hyperhomocysteinemia with an increased incidence of cardiovascular disease events has been reported among stable renal transplant recipients (RTRs). Preliminary studies in a small number of these individuals have shown that serum creatinine and cystatin C, both markers of kidney function and glomerular filtration rate, are independent determinants of fasting tHcy concentrations; however, determinants of tHcy concentrations after a methionine load have not been studied. METHODS: We determined the prevalence of both fasting and 4-h post-methionine load (PML) tHcy concentrations in 78 stable RTRs and compared the role of cystatin C with the role of serum creatinine as determinants of fasting and PML tHcy. RESULTS: Of the 78 RTRs, 21 (26.9%) had fasting and PML tHcy within the respective reference intervals, and 57 (73.1%) had increased plasma tHcy. Of these 57 RTRs, 22 had fasting hyperhomocysteinemia, 9 had PML hyperhomocysteinemia, and 26 had combined hyperhomocysteinemia (both fasting and PML). Unadjusted Pearson correlations showed that fasting plasma tHcy correlated with both cystatin C (r = 0.564; P <0.001) and creatinine (r = 0.519; P <0.001) and that increases in PML tHcy modestly correlated with cystatin (r = 0.205; P = 0.072), but not creatinine (r = 0.057; P = 0.624). General linear regression modeling with stepwise analysis of covariance showed that both cystatin C (partial R = 0.554; P <0.001) and creatinine (partial R = 0.535; P <0.001) were independent predictors of fasting tHcy, but of the two, only cystatin C (partial R = 0.242; P = 0.035) was an independent predictor of increased PML tHcy. CONCLUSIONS: Clinically stable RTRs have an excess prevalence of moderate hyperhomocysteinemia, and additional cases can be detected by methionine loading. Both creatinine and cystatin C are independent predictors of fasting tHcy in these individuals; however, only cystatin C is a determinant of tHcy concentration after a methionine load, probably because cystatin C is a more sensitive marker of glomerular filtration rate than serum creatinine. PMID- 11427459 TI - Supersensitive time-resolved immunofluorometric assay of free prostate-specific antigen with nanoparticle label technology. AB - BACKGROUND: The extreme specific activity of the long-lifetime fluorescent europium(III) chelate nanoparticles and the enhanced monovalent binding affinity of multivalent nanoparticle-antibody bioconjugates are attractive for noncompetitive immunoassay. METHODS: We used a noncompetitive, two-step immunoassay design to measure free prostate-specific antigen (PSA). Europium(III) chelate nanoparticles (107 nm in diameter) were coated with a monoclonal anti-PSA antibody (intrinsic affinity, 6 x 10(9) L/mol). The nanoparticle-antibody bioconjugates had an average of 214 active binding sites per particle and a monovalent binding affinity of 7 x 10(10) L/mol. The assay was performed in a low fluorescence microtitration well passively coated with an another monoclonal anti PSA antibody (affinity, 2 x 10(10) L/mol), and the europium(III) fluorescence was measured directly from the bottom of the well by a standard time-resolved microtitration plate fluorometer. RESULTS: The detection limit (mean + 2 SD) was 0.040 ng/L (7.3 x 10(5) molecules/mL), and the dynamic detection range covered four orders of magnitude in a 3-h total assay time. The imprecision (CV) over the whole assay range was 2-10%. The detection limit of the assay was limited by the fractional nonspecific binding of the bioconjugate to the solid phase (0.05%), which was higher than the nonspecific binding of the original antibody (<0.01%). CONCLUSIONS: The sensitivity of the new assay is equal to that of the ambient analyte, microspot immunoassay and will be improved by use of optimized, high binding-site density nanoparticle-antibody bioconjugates with reduced nonspecific binding and improved monovalent binding affinity. PMID- 11427460 TI - Near-infrared spectroscopy for measuring urea in hemodialysis fluids. AB - BACKGROUND: Near-infrared spectroscopy is proposed as a method for providing real time urea concentrations during hemodialysis treatments. The feasibility of such noninvasive urea measurements is evaluated in undiluted dialysate fluid. METHODS: Near-infrared spectra were collected from calibration solutions of urea prepared in dialysate fluid. Spectra were collected over three distinct spectral regions, and partial least-squares calibration models were optimized and compared for each. Selectivity for urea was demonstrated with two-component samples composed of urea and glucose in the dialysate matrix. The clinical significance of this approach was assessed by measuring urea in real hemodialysate samples. RESULTS: Urea absorptions within the combination and short-wavelength, near-infrared spectral regions provided sufficient spectral information for sound calibration models in the dialysate matrix. The combination spectral region had SEs of calibration (SEC) and prediction (SEP) of 0.38 mmol/L and 0.26 mmol/L, respectively, over the 4720-4600 cm(-1) spectral range with 5 partial least square factors. A second calibration model was established over the combination region from a series of solutions prepared with independently variable concentrations of urea and glucose. The best calibration model for urea in the presence of variable glucose concentrations had a SEC of 0.6 mmol/L and a SEP of 0.4 mmol/L for a 5-factor model over the 4600-4350 cm(-1) spectral range. There was no significant decrease in SEP when the 4720-4600 cm(-1) calibration model was used to measure urea in real samples collected during actual hemodialysis. CONCLUSIONS: Urea can be determined with sufficient sensitivity and selectivity for clinical measurements within the matrix of the hemodialysis fluid. PMID- 11427461 TI - Infrared analysis of urinary calculi by a single reflection accessory and a neural network interpretation algorithm. AB - BACKGROUND: Preparation of KBr tablets, used for Fourier transform infrared (FT IR) analysis of urinary calculus composition, is time-consuming and often hampered by pellet breakage. We developed a new FT-IR method for urinary calculus analysis. This method makes use of a Golden Gate Single Reflection Diamond Attenuated Total Reflection sample holder, a computer library, and an artificial neural network (ANN) for spectral interpretation. METHODS: The library was prepared from 25 pure components and 236 binary and ternary mixtures of the 8 most commonly occurring components. The ANN was trained and validated with 248 similar mixtures and tested with 92 patient samples, respectively. RESULTS: The optimum ANN model yielded root mean square errors of 1.5% and 2.3% for the training and validation sets, respectively. Fourteen simple expert rules were added to correct systematic network inaccuracies. Results of 92 consecutive patient samples were compared with those of a FT-IR method with KBr tablets, based on an initial computerized library search followed by visual inspection. The bias was significantly different from zero for brushite (-0.8%) and the concomitantly occurring whewellite (-2.8%) and weddellite (3.8%), but not for ammonium hydrogen urate (-0.1%), carbonate apatite (0.5%), cystine (0.0%), struvite (0.4%), and uric acid (-0.1%). The 95% level of agreement of all results was 9%. CONCLUSIONS: The new Golden Gate method is superior because of its smaller sample size, user-friendliness, robustness, and speed. Expert knowledge for spectral interpretation is minimized by the combination of a library search and ANN prediction, but visual inspection remains necessary. PMID- 11427462 TI - Tumor necrosis factor-alpha in serum of patients with inflammatory bowel disease as measured by a highly sensitive immuno-PCR. AB - BACKGROUND: The significance of serum concentrations of tumor necrosis factor alpha (TNF-alpha) in the pathogenesis of inflammatory bowel disease (IBD) is uncertain. We measured TNF-alpha in serum from IBD patients by immuno-PCR to analyze the relationship between TNF-alpha and pathophysiologic state in IBD. METHODS: Serum samples were collected from 54 healthy blood donors, 29 patients with ulcerative colitis (UC; 46 samples), and 7 patients with Crohn disease (CD; 8 samples). DNA label was generated by PCR amplification using biotinylated primer and was bound with streptavidin to biotinylated third antibody. TNF-alpha sandwiched by antibodies was detected by PCR amplification of the DNA label. RESULTS: TNF-alpha could be measured in all samples. The median serum concentration in IBD patients overall was approximately 390-fold higher than in healthy donors (median increase, 380-fold for UC, 640-fold for CD). The median serum TNF-alpha concentration was 1.7-fold higher in the active stage of UC than in the inactive stage (P <0.05), and this difference could be detected in individual patients. CONCLUSIONS: Sensitive measurement of serum TNF-alpha could provide an important pathophysiologic marker for the presence and activity of IBD. PMID- 11427463 TI - A novel test for the measurement of skin cholesterol. PMID- 11427464 TI - Chlamydia pneumoniae seropositivity and hyperhomocysteinemia are linked in patients with atherosclerosis. PMID- 11427465 TI - One-step solid-phase extraction procedure for F(2)-isoprostanes. PMID- 11427466 TI - Electrospray mass spectrometry: an efficient method to detect silent hemoglobin variants causing erythrocytosis. PMID- 11427467 TI - Quantification of aromatic hydrocarbon receptor (AHR) and related genes by calibrated reverse transcription-PCR in blood mononuclear cells. PMID- 11427468 TI - Biomarkers of bone turnover after a short period of steroid therapy in elderly men. PMID- 11427469 TI - HPLC assay with ultraviolet detection for therapeutic drug monitoring of sirolimus. PMID- 11427470 TI - Determination of D-mannose in serum by capillary electrophoresis. PMID- 11427471 TI - Quantification of reduced and oxidized glutathione in whole blood samples by capillary electrophoresis. PMID- 11427472 TI - Rapid screening for KIT mutations by capillary electrophoresis. PMID- 11427473 TI - 2-Mercaptoethanol treatment improves measurement of an IgMkappa M-protein by capillary electrophoresis. PMID- 11427474 TI - Influence quantities and uncertainty of measurement. PMID- 11427475 TI - Serendipitous detection of umbilical venous catheter displacement by cardiac troponin I measurement. PMID- 11427476 TI - How to improve total error modeling by accounting for error sources beyond imprecision and bias. PMID- 11427477 TI - Limitations of genotyping based on amplicon melting temperature. PMID- 11427478 TI - Use of heterophilic antibody blocking agent (HBT) in reducing false-positive hCG results. PMID- 11427481 TI - Ribosome-inactivating proteins from plants: more than RNA N-glycosidases? AB - Many plants contain proteins that are capable of inactivating ribosomes and accordingly are called ribosome-inactivating proteins or RIPs. These typical plant proteins receive a lot of attention in biological and biomedical research because of their unique biological activities toward animal and human cells. In addition, evidence is accumulating that some RIPs play a role in plant defense and hence can be exploited in plant protection. To understand the mode of action of RIPs and to optimize their medical and therapeutical applications and their use as antiviral compounds in plant protection, intensive efforts have been made to unravel the enzymatic activities of RIPs and provide a structural basis for these activities. Though marked progress has been made during the last decade, the enzymatic activity of RIPs has become a controversial issue because of the concept that RIPs possess, in addition to their classical RNA N-glycosidase and polynucleotide:adenosine glycosidase activity, other unrelated enzymatic activities. Moreover, the presumed novel enzymatic activities, especially those related to diverse nuclease activities, are believed to play an important role in various biological activities of RIPs. However, both the novel enzymatic activities and their presumed involvement in the biological activities of RIPs have been questioned because there is evidence that the activities observed are due to contaminating enzymes. We offer a critical review of the pros and cons of the putative novel enzymatic activities of RIPs. Based on the available data, it is suggested that there is little conclusive evidence in support of the presumed activities and that in the past too little attention has been given to the purity of the RIP preparation. The antiviral activity and mode of action of RIPs in plants are discussed in view of their classical and presumed novel enzymatic activities. PMID- 11427482 TI - The role of c-Myb and Sp1 in the up-regulation of methionine adenosyltransferase 2A gene expression in human hepatocellular carcinoma. AB - Liver-specific and non-liver-specific methionine adenosyltransferase (MAT) are products of two genes, MAT1A and MAT2A, respectively, that catalyze the formation of S-adenosylmethionine. We showed a switch from MAT1A to MAT2A expression at the transcriptional level in human hepatocellular carcinoma (HCC) that facilitates cancer cell growth. The purpose of the present study was to better understand the molecular mechanism of increased MAT2A expression in HCC. In vitro DNase I footprinting analysis revealed two protected sites (-354 to -312 and -73 to -28) using nuclear proteins from HCC and HepG2 cells, but not normal liver. These sites are also protected in HepG2 cells on in vivo DNase I footprinting analysis. These protected sites contain consensus binding sites for c-Myb and Sp1. In HCC, the mRNA levels of c-myb and Sp1 and binding to their respective sites increased. Mutation of the c-Myb or Sp1 site reduced MAT2A promoter activity by 67% and 50%, respectively. The importance of these cis-acting elements and trans-activating factors was confirmed using heterologous promoter and expression vectors. Increased expression of c-Myb and Sp1 and binding to the MAT2A promoter contribute to transcriptional up-regulation of MAT2A in HCC.-Yang, H., Huang, Z. Z., Wang, J., Lu, S. C. The role of c-Myb and Sp1 in the up-regulation of methionine adenosyltransferase 2A gene expression in human hepatocellular carcinoma. PMID- 11427483 TI - Activation of cellular invasion by trefoil peptides and src is mediated by cyclooxygenase- and thromboxane A2 receptor-dependent signaling pathways. AB - We have investigated the possible functional relationships between cellular invasion pathways induced by trefoil factors (TFFs), src, and the cyclooxygenases COX-1 and COX-2. Pharmacological inhibitors of the Rho small GTPase (C3 exoenzyme), phospholipase C (U-73122), cyclooxygenases (SC-560, NS-398), and the thromboxane A2 receptor (TXA2-R) antagonist SQ-295 completely abolished invasion induced by intestinal trefoil factor, pS2, and src in kidney and colonic epithelial cells MDCKts.src and PCmsrc. In contrast, invasion was induced by the TXA2-R mimetic U-46619, constitutively activated forms of the heterotrimeric G proteins Galphaq (AGalphaq), Galpha12, Galpha13 (AGalpha12/13), which are signaling elements downstream of TXA2-R. Ectopic overexpression of pS2 cDNA and protein in MDCKts.src-pS2 cells and human colorectal cancer cells HCT8/S11-pS2 initiate distinct invasion signals that are Rho independent and COX and TXA2-R dependent. We detected a marked induction of COX-2 protein and accumulation of the stable PGH2/TXA2 metabolite TXB2 in the conditioned medium from cells transformed by src. This led to activation of the TXA2-R-dependent invasion pathway, which is monitored via a Rho- and Galpha12/Galpha13-independent mechanism using the Galphaq/PKC signaling cascade. These findings identify a new intracrine/paracrine loop that can be monitored by TFFs and src in inflammatory diseases and progression of colorectal cancers. PMID- 11427484 TI - A cutaneous gene therapy approach to human leptin deficiencies: correction of the murine ob/ob phenotype using leptin-targeted keratinocyte grafts. AB - Leptin deficiency produces a phenotype of obesity, diabetes, and infertility in the ob/ob mouse. In humans, leptin deficiency occurs in some cases of congenital obesity and in lipodystrophic disorders characterized by reduced adipose tissue and insulin resistance. Cutaneous gene therapy is considered an attractive potential method to correct circulating protein deficiencies, since gene transferred human keratinocytes can produce and secrete gene products with systemic action. However, no studies showing correction of a systemic defect have been reported. We report the successful correction of leptin deficiency using cutaneous gene therapy in the ob/ob mouse model. As a feasibility approach, skin explants from transgenic mice overexpressing leptin were grafted on immunodeficient ob/ob mice. One month later, recipient mice reached body weight values of lean animals. Other biochemical and clinical parameters were also normalized. In a second human gene therapy approach, a retroviral vector encoding both leptin and EGFP cDNAs was used to transduce HK and, epithelial grafts enriched in high leptin-producing HK were transplanted to immunosuppressed ob/ob mice. HK-derived leptin induced body weight reduction after a drop in blood glucose and food intake. Leptin replacement through genetically engineered HK grafts provides a valuable therapeutic alternative for permanent treatment of human leptin deficiency conditions. PMID- 11427485 TI - Down-regulation of L-type calcium channel in pups born to 52 kDa SSA/Ro immunized rabbits. AB - Congenital heart block is considered a model of passively acquired autoimmune disease in which the mother generates anti-SSA/Ro and/or anti-SSB/La antibodies that cross the placenta and presumably injure the heart of developing fetus. CHB is accompanied by ECG abnormalities including AV block, sinus bradycardia, and ventricular dysfunction. Our previous data indicate that these abnormalities are caused by maternal autoantibody-mediated disturbance of L-type Ca channels. To investigate the consequence of chronic exposure of L-type Ca channels in newborn pups to maternal autoantibodies during pregnancy, we immunized female rabbits with human 52 kDa-SSA/Ro (Ro52) recombinant protein. ECG revealed that pups from the immunized group had varying degrees of conduction defects. In addition, I(CaL) density and protein were reduced in hearts of pups from the immunized group. Sera and purified IgG from immunized rabbits inhibited I(Ba) recorded from oocytes with expressed alpha(1C) and beta(2a) subunits of L-type Ca channel. Pups born to Ro52 immunized mothers exhibited down-regulation of L-type calcium channels in heart. The data provide new insight into the pathogenesis of congenital heart block. PMID- 11427486 TI - Heparan mimetic regulates collagen expression and TGF-beta1 distribution in gamma irradiated human intestinal smooth muscle cells. AB - Radiation-induced intestinal fibrosis is characterized by collagen accumulation, a process in which TGF-beta1 plays a key role. We analyzed the effects of gamma radiation on collagen expression and TGF-beta1 distribution in human intestinal smooth muscle cells (HISM). We investigated the activity of a carboxymethylated and sulfated dextran (RG-1503), exhibiting antifibrotic properties and promoting in vivo intestinal tissue repair, on irradiated HISM. After (60)Co irradiation (10 Gy), HISM were labeled with [(3)H] proline (+/-RG-1503). Radiolabeled collagen I, III, and V were quantified by SDS-PAGE. TGF-beta1 was quantified by ELISA in culture medium, pericellular and intracellular compartments. Irradiation induced a specific 2.85-fold increase in collagen III production by HISM. Collagen V decreased by 80% 72 h after irradiation. Pericellular TGF-beta1 was increased (up to twofold) in irradiated HISM. RG-1503 added before or after irradiation reversed both mRNA and protein levels of collagen III and V to control values. RG-1503 decreased the amount of TGF-beta1 in the cell layer below the control values. Irradiation of HISM induced the development of a fibrotic phenotype in terms of collagen production and TGF-beta1 distribution. The antifibrotic RG-1503 restored HISM physiological characteristics and may represent a promising therapeutic approach for radiation-induced intestinal fibrosis. PMID- 11427487 TI - ATP binding cassette transporter ABCA1 modulates the secretion of apolipoprotein E from human monocyte-derived macrophages. AB - Apolipoprotein E (apoE) produced by macrophages in the arterial wall protects against atherosclerosis, but the regulation of its secretion by these cells is poorly understood. Here we investigated the contribution of the adenosine triphosphate binding cassette transporters ABCA1 and ABC8 to the secretion of apoE from either primary human monocyte-derived macrophages (HMDM) or human THP1 macrophages. During incubations of up to 6 h, apoE secretion from both THP1 macrophages and HMDM was stimulated by 8-Br-cAMP, which activates ABCA1 expression. The putative ABCA1 inhibitor glyburide and antisense oligonucleotides directed against ABCA1 mRNA significantly reduced apoE secretion from THP1 macrophages and HMDM. Antisense oligonucleotides directed against ABC8 mRNA also inhibited apoE secretion, although this inhibition was less pronounced and consistent than in the case of ABCA1. ApoE secretion from HMDM of ABCA1-deficient patients with Tangier disease was also decreased. ApoE mRNA expression was not affected by inhibition of ABCA1 or ABC8 in normal HMDM or the lack of functional ABCA1 in HMDM from Tangier disease patients. Inhibition of ABCA1 in HMDM prevented the occurrence of anti-apoE-immunoreactive granular structures in the plasma membrane. We conclude that ABCA1 and, to a lesser extent, ABC8 both promote secretion of apoE from human macrophages. PMID- 11427488 TI - Metabolic radiolabeling: experimental tool or Trojan horse? (35)S-Methionine induces DNA fragmentation and p53-dependent ROS production. AB - Despite the general assumption that widely used radiolabeled metabolites such as [(35)S]methionine and (3)H-thymidine do not adversely affect or perturb cell function, we and others have shown that such low-energy beta-emitters can cause cell cycle arrest and apoptosis of proliferating cells. The goal of the present study was to elucidate the targets and mechanisms of [(35)S]methionine-induced cellular toxicity. Comet analyses (single-cell electrophoresis) demonstrated dose dependent DNA fragmentation in rabbit smooth muscle cells within a time frame (1 4 h) well within that of most radiolabeling protocols, whereas fluorescence analyses using a peroxide/hydroperoxide-sensitive dye revealed production of reactive oxygen species (ROS). Although ROS generation was inhibitable by antioxidants, DNA fragmentation was not inhibited and was in fact observed even under hypoxic conditions, suggesting that beta-radiation-induced DNA damage can occur independently of ROS formation. Studies with p53(+/+) and p53(-/-) human colorectal carcinoma cells further demonstrated the dissociation of early DNA damage from ROS formation in that both cell types exhibited DNA fragmentation in response to radiolabeling whereas only the p53(+/+) cells exhibited significant increases in ROS formation, which occurred well after significant DNA damage was observed. These findings demonstrate that metabolically incorporated low-energy beta-emitters such as [(35)S]methionine and (3)H-thymidine can induce DNA damage, thereby initiating cellular responses leading to cell cycle arrest or apoptosis. The results of this study require a reevaluation using low-energy beta-emitters to follow not only experimental protocols in vivo processes, but also acceptable exposure levels of these genotoxic compounds in the workplace and environment. PMID- 11427489 TI - NAD(P)H, a directly operating antioxidant? AB - Endogenous oxygen- and nitrogen-centered free radicals are considered to play a decisive role in a variety of diseases such as neurodegenerative disorders, atherosclerosis, or cancer. Directly operating antioxidants limit the action of freely diffusing radicals by scavenging the attacking, oxidizing radical and re reducing the oxidized biomolecule, i.e., the biomolecule-derived radical. From textbooks of biochemistry it is understood that NAD(P)H acts as a hydride (hydrogen anion) donor in a variety of enzymatic processes. One example is the re reduction of GSSG to GSH, catalyzed by glutathione reductase. Because of this reaction, NADPH has been suggested to also act as an indirectly operating antioxidant, thus maintaining the antioxidative power of glutathione. To the best of our knowledge, however, neither NADPH nor NADH has been considered to be directly operating antioxidants. Based on recently published data, new experiments, and theoretical considerations, we propose that NAD(P)H represents a decisive, directly operating antioxidant that should be considered of major importance in the mitochondrial compartment. NAD(P)H fulfills this task both by scavenging toxic free radicals and repairing biomolecule-derived radicals. PMID- 11427490 TI - Mind over immunity. AB - The central nervous system regulates the innate immune system by elaborating anti inflammatory hormone cascades in response to bacterial products and immune mediators. We recently discovered that the central nervous system also responds via acetylcholine-mediated efferent signals carried through the vagus nerve. Nicotinic cholinergic receptors expressed on macrophages detect these signals and respond with a dampened cytokine response. Vagus nerve stimulators can mimic this response and can prevent lethal endotoxemia. This newly appreciated cholinergic anti-inflammatory pathway provides a neural substrate to study brain-immune interactions and might be harnessed for therapy of cytokine-mediated disease. PMID- 11427491 TI - Protein kinase C-mediated calcium entry dependent upon dihydropyridine sensitive channels: a T cell receptor-coupled signaling pathway involved in IL-4 synthesis. PMID- 11427492 TI - Trans-plasma membrane electron transport induces macrophage-mediated low density lipoprotein oxidation. PMID- 11427493 TI - Oxidative stress-induced activation of Lyn recruits sphingomyelinase and is requisite for its stimulation by Ara-C. PMID- 11427494 TI - Regulation of spontaneous glutamate release activity in osteoblastic cells and its role in differentiation and survival: evidence for intrinsic glutamatergic signaling in bone. PMID- 11427495 TI - Caloric restriction decreases mitochondrial free radical generation at complex I and lowers oxidative damage to mitochondrial DNA in the rat heart. PMID- 11427496 TI - The lysosomal protease cathepsin D mediates apoptosis induced by oxidative stress. PMID- 11427497 TI - Tumor necrosis factor alpha attenuates interferon alpha signaling in the liver: involvement of SOCS3 and SHP2 and implication in resistance to interferon therapy. PMID- 11427498 TI - Analysis of the structure and function relationship of the human apolipoprotein E in vivo, using adenovirus-mediated gene transfer. PMID- 11427499 TI - Selective inhibition of CTL activation by a dipalmitoyl-phospholipid that prevents the recruitment of signaling molecules to lipid rafts. PMID- 11427500 TI - Carnitine protects the molecular chaperone activity of lens alpha-crystallin and decreases the post-translational protein modifications induced by oxidative stress. PMID- 11427501 TI - Antiproliferative action of dopamine and norepinephrine in neuroblastoma cells expressing the human dopamine transporter. PMID- 11427502 TI - Paracrinally stimulated expansion of early human hemopoietic progenitors by stroma-generated cyclic ADP-ribose. PMID- 11427503 TI - Resveratrol, a tumor-suppressive compound from grapes, induces apoptosis via a novel mitochondrial pathway controlled by Bcl-2. PMID- 11427504 TI - Administration of NF-kappaB decoy inhibits pancreatic activation of NF-kappaB and prevents diabetogenesis by alloxan in mice. PMID- 11427505 TI - Up-regulation of the fibrogenic cytokine TGF-beta1 by oxysterols: a mechanistic link between cholesterol and atherosclerosis. PMID- 11427506 TI - Celecoxib loses its anti-inflammatory efficacy at high doses through activation of NF-kappaB. PMID- 11427507 TI - Down-regulation of amphiphysin-1 is responsible for reduced receptor-mediated endocytosis in the senescent cells. PMID- 11427508 TI - Circulating hematopoietic stem cells serve as novel targets for in utero gene therapy. PMID- 11427509 TI - Syndecan-4 up-regulation in proliferative renal disease is related to microfilament organization. PMID- 11427510 TI - Diffusion-limited translocation mechanism of protein kinase C isotypes. PMID- 11427511 TI - Use of an anaerobic environment to preserve the endogenous activity of protein tyrosine phosphatases isolated from intact cells. PMID- 11427512 TI - Alcohol consumption is controlled by angiotensin II. PMID- 11427513 TI - Interaction of disintegrins with human neutrophils induces cytoskeleton reorganization, focal adhesion kinase activation, and extracellular-regulated kinase-2 nuclear translocation, interfering with the chemotactic function. PMID- 11427514 TI - Protein and peptide delivery via engineered polyomavirus-like particles. PMID- 11427515 TI - Octanol antagonism of ethanol teratogenesis. PMID- 11427516 TI - Trp1-dependent enhancement of salivary gland fluid secretion: role of store operated calcium entry. PMID- 11427517 TI - Dystrophin-deficient cardiomyocytes are abnormally vulnerable to mechanical stress-induced contractile failure and injury. PMID- 11427518 TI - Interaction of Clostridium botulinum C2 toxin with lipid bilayer membranes and Vero cells: inhibition of channel function by chloroquine and related compounds in vitro and intoxification in vivo. PMID- 11427519 TI - Heparanase as mediator of angiogenesis: mode of action. PMID- 11427520 TI - Substrate analogs incorporating beta-amino acids: potential application for peptidase inhibition. PMID- 11427521 TI - Vascular endothelial growth factor induces cyclooxygenase-dependent proliferation of endothelial cells via the VEGF-2 receptor. PMID- 11427522 TI - A novel group IIA phospholipase A2 interacts with v-Src oncoprotein from RSV transformed hamster cells. AB - We have isolated a novel isoform of phospholipase A(2). This enzyme was designated srPLA(2) because it was discovered while analyzing the proteins interacting with different forms of the v-Src oncoproteins isolated from Rous sarcoma virus-transformed hamster cells. It contains all the functional regions of the PLA(2) group IIA proteins but differs at its C-terminal end where there is an additional stretch of 8 amino acids. The SrPLA(2) isoform was detected as a 17 kDa precursor in cells and as a mature 14-kDa form secreted in culture medium. A direct interaction of the 17-kDa precursor with the Src protein was observed in lysates of transformed cells. Both the 17- and 14-kDa forms were found to be phosphorylated on tyrosine. To our knowledge, this is the first report of a PLA(2) group II protein that is tyrosine phosphorylated. We surmise that srPLA(2) interacts with the Src protein at the cell membrane during the process of its maturation. PMID- 11427523 TI - Binding of microsomal triglyceride transfer protein to lipids results in increased affinity for apolipoprotein B: evidence for stable microsomal MTP-lipid complexes. AB - Apolipoprotein B (apoB) and microsomal triglyceride transfer protein (MTP) are known to interact with each other. We evaluated the effect of different lipids on the protein-protein interactions between MTP and apoB100 or its C-terminally truncated forms. Negatively charged lipids decreased protein-protein interactions between apoB and MTP. In contrast, zwitterionic phospholipids enhanced (2-4-fold) the binding of apoB100 to MTP by increasing affinity (1.5-3-fold) between these proteins without affecting the number of binding sites. Similarly, phospholipids augmented (1.5-4-fold) the binding of various C-terminally truncated apoB peptides to MTP. The increased binding was greater for apoB peptides containing lipid-binding domains, such as apoB28 and apoB42. Surprisingly, preincubation of apoB28 with lipid vesicles had no effect on MTP binding. In contrast, incubation of MTP with lipid vesicles resulted in a stable association of MTP with vesicles, and MTP-lipid vesicles bound better (5-fold increase) to LDL than did lipid-free MTP. To determine whether MTP exists stably associated with lipids in cells, microsomal contents from COS cells expressing MTP, HepG2 cells, and mouse liver were ultracentrifuged, and MTP was visualized in different density fractions. MTP was found associated and unassociated with lipids. In contrast, apoB17 and apoB:270-570 were present unassociated with lipids in COS cells. These studies show that the binding of MTP to lipids results in increased affinity for apoB and that stable MTP-lipid complexes exist in the lumen of the endoplasmic reticulum. Protein-protein interactions between apoB and MTP may juxtapose lipids associated with MTP to lipid-binding domains of apoB and facilitate hydrophobic interactions leading to enhance affinity. We speculate that MTP-lipid complexes may serve as nuclei to form "primordial lipoproteins" and may also play a role in the bulk addition of lipids during the "core expansion" of these lipoproteins. PMID- 11427524 TI - Soluble Jagged 1 represses the function of its transmembrane form to induce the formation of the Src-dependent chord-like phenotype. AB - We have previously demonstrated that the expression of the soluble extracellular domain of the transmembrane ligand for Notch receptors, Jagged 1 (sJ1), in NIH 3T3 cells results in the formation of a matrix-dependent chord-like phenotype, the loss of contact inhibition of growth, and an inhibition of pro-alpha 1(I) collagen expression. In an effort to define the mechanism by which sJ1 induces this phenotype, we report that sJ1 transfectants display biochemical and cytoskeletal alterations consistent with the activation of Src. Indeed, cotransfection of sJ1 transfectants with a dominant-negative mutant of Src resulted in the loss of matrix-dependent chord formation and correlated with the restoration of type I collagen expression and contact inhibition of growth. We also report that the sJ1-mediated induction of Src activity and related phenotypes, including chord formation, may result from the inhibition of endogenous Jagged 1-mediated Notch signaling since it was not possible to detect an sJ1-dependent induction of CSL-dependent transcription in these cells. Interestingly, NIH 3T3 cells transfected with dominant-negative (but not constitutively active) mutants of either Notch 1 or Notch 2 displayed a similar Src-related phenotype as the sJ1 transfectants. These data suggest that the ability of sJ1 to mediate chord formation is Src-dependent and requires the repression of endogenous Jagged 1-mediated Notch signaling, which is tolerant to the destabilization of the actin cytoskeleton, a mediator of cell migration. PMID- 11427525 TI - Remodeling of Kv4.3 potassium channel gene expression under the control of sex hormones. AB - Kv4.3 channels are important molecular components of transient K(+) currents (Ito currents) in brain and heart. They are involved in setting the frequency of neuronal firing and heart pacing. Altered Kv4.3 channel expression has been demonstrated under pathological conditions like heart failure indicating their critical role in heart function. Thyroid hormone studies suggest that their expression in the heart may be hormonally regulated. To explore the possibility that sex hormones control Kv4.3 expression, we investigated whether its expression changes in the pregnant uterus. This organ represents a unique model to study Ito currents, because it possesses this type of K(+) current and undergoes dramatic changes in function and excitability during pregnancy. We cloned Kv4.3 channel from myometrium and found that its protein and transcript expression is greatly diminished during pregnancy. Experiments in ovariectomized rats demonstrate that estrogen is one mechanism responsible for the dramatic reduction in Kv4.3 expression and function prior to parturition. Furthermore, the reduction of plasma membrane Kv4.3 protein is accompanied by a perinuclear localization suggesting that cell trafficking is also controlled by sex hormones. Thus, estrogen remodels the expression of Kv4.3 in myometrium by directly diminishing its transcription and, indirectly, by altering Kv4.3 delivery to the plasma membrane. PMID- 11427526 TI - Phospholipase D is required in the signaling pathway leading to p38 MAPK activation in neutrophil-like HL-60 cells, stimulated by N-formyl-methionyl leucyl-phenylalanine. AB - Human acute myelogenous leukemia cells (HL-60 cells) can be induced to differentiate to neutrophils by exposure to dibutyryl-cyclic AMP. The differentiation of HL-60 cells allowed the mitogen-activated protein kinases p38 and p44/p42 to be rapidly and transiently activated upon stimulation with N formyl-methionyl-leucyl-phenylalanine (fMLP). Western blot analysis using phosphospecific p38 and p44/p42 mitogen-activated protein kinase antibodies showed that increasing concentrations of ethanol or 1-butanol but not 2-butanol (0.05-0.5%) inhibited fMLP-induced p38 activation but did not inhibit p44/p42 activation. These data indicated that activation of phospholipase D (PLD) was required for activation of p38 but not p44/p42. We compared the effect of fMLP with those of tumor necrosis factor alpha (TNF alpha) and granulocyte-macrophage colony-stimulating factor (GM-CSF). We found that ethanol did not inhibit p38 phosphorylation upon stimulation with either GM-CSF or TNF alpha. These results suggested that in cells stimulated with fMLP, PLD was upstream of p38. To further test the involvement of PLD, we used antisense inhibition of human PLD1 expression. Treatment with antisense oligonucleotides inhibited p38 but not p44/p42 phosphorylation. These data supported a role for human PLD1 in fMLP induced p38 activation in neutrophil-like HL-60 cells. In addition, the results obtained with TNF alpha and GM-CSF demonstrated that p38 activation occurred independently of PLD activation. PMID- 11427527 TI - A novel approach to the design of inhibitors of human secreted phospholipase A2 based on native peptide inhibition. AB - Human Type IIA secreted phospholipase A(2) (sPLA(2)-IIA) is an important modulator of cytokine-dependent inflammatory responses and a member of a growing superfamily of structurally related phospholipases. We have previously shown that sPLA(2)-IIA is inhibited by a pentapeptide sequence comprising residues 70-74 of the native sPLA(2)-IIA protein and that peptides derived from the equivalent region of different sPLA(2)-IIA species specifically inhibit the enzyme from which they are derived. We have now used an analogue screen of the human pentapeptide (70)FLSYK(74) in which side-chain residues were substituted, together with molecular docking approaches that modeled low-energy conformations of (70)FLSYK(74) bound to human sPLA(2)-IIA, to generate inhibitors with improved potency. Importantly, the modeling studies showed a close association between the NH(2) and COOH termini of the peptide, predicting significant enhancement of the potency of inhibition by cyclization. Cyclic compounds were synthesized and indeed showed 5-50-fold increased potency over the linear peptide in an Escherichia coli membrane assay. Furthermore, the potency of inhibition correlated with steady-state binding of the cyclic peptides to sPLA(2)-IIA as determined by surface plasmon resonance studies. Two potential peptide interaction sites were identified on sPLA(2)-IIA from the modeling studies, one in the NH(2)-terminal helix and the other in the beta-wing region, and in vitro association assays support the potential for interaction of the peptides with these sites. The inhibitors were effective at nanomolar concentrations in blocking sPLA(2)-IIA-mediated amplification of cytokine-induced prostaglandin synthesis in human rheumatoid synoviocytes in culture. These studies provide an example where native peptide sequences can be used for the development of potent and selective inhibitors of enzyme function. PMID- 11427528 TI - A new lysozyme fold. Crystal structure of the muramidase from Streptomyces coelicolor at 1.65 A resolution. AB - Cellosyl is a bacterial muramidase from Streptomyces coelicolor. Similar to other lysozymes, the enzyme cleaves the beta-1,4-glycosidic bond between N acetylmuramic acid and N-acetylglucosamine units, but it also exhibits a beta-1,4 N,6-O-diacetylmuramidase activity. The latter enables Cellosyl to degrade the cell walls of Staphylococcus aureus, which are not hydrolyzed by chicken-, goose , or bacteriophage T4-type lysozymes. The enzymatic activity and amino acid sequence of Cellosyl group it with lysozymes of the Chalaropsis type, for which no detailed structural information has been available so far. The crystal structure of Cellosyl from S. coelicolor has been determined to a resolution of 1.65 A and refined to an R-factor of 15.2%. The enzyme is comprised of a single domain and possesses an unusual beta/alpha-barrel fold. The last strand, beta 8, of the (beta/alpha)(5)beta(3)-barrel is found to be antiparallel to strands beta 7 and beta 1. Asp-9, Asp-98, and Glu-100 are located at the active site. The structure of Cellosyl exhibits a new lysozyme fold and represents a new class of polysaccharide-hydrolyzing beta/alpha-barrels. PMID- 11427529 TI - The interaction of DNA mismatch repair proteins with human exonuclease I. AB - Exonucleolytic degradation of DNA is an essential part of many DNA metabolic processes including DNA mismatch repair (MMR) and recombination. Human exonuclease I (hExoI) is a member of a family of conserved 5' --> 3' exonucleases, which are implicated in these processes by genetic studies. Here, we demonstrate that hExoI binds strongly to hMLH1, and we describe interaction regions between hExoI and the MMR proteins hMSH2, hMSH3, and hMLH1. In addition, hExoI forms an immunoprecipitable complex with hMLH1/hPMS2 in vivo. The study of interaction regions suggests a biochemical mechanism of the involvement of hExoI as a downstream effector in MMR and/or DNA recombination. PMID- 11427530 TI - Functional characterization of mutants in the predicted pore region of the rabbit cardiac muscle Ca(2+) release channel (ryanodine receptor isoform 2). AB - A highly conserved amino acid sequence, GVRAGGGIGD(4831), which may form part of the Ca(2+) release channel pore in RyR2, was subjected to Ala scanning or Ala to Val mutagenesis; function was then measured by expression in HEK-293 cells, followed by Ca(2+) photometry, high affinity [(3)H]ryanodine binding, and single channel recording. All mutants except I4829A and I4829T (corresponding to the I4897T central core disease mutant in RyR1) displayed caffeine-induced Ca(2+) release in HEK-293 cells; only mutants G4826A, I4829V, and G4830A retained high affinity [(3)H]ryanodine binding; and single-channel function was found for all mutants tested, except for G4822A and A4825V. EC(50) values for caffeine-induced Ca(2+) release were increased for G4822A, R4824A, G4826A, G4828A, and D4831A; decreased for V4823A; and unchanged for A4825V, G4827A, I4829V, and G4830A. Ryanodine (10 microm), which did not stimulate Ca(2+) release in wild type (wt), did so in Ala mutants in amino acids 4823-4827. It inhibited the caffeine response in wt and most mutants, but enhanced the amplitude of caffeine-induced Ca(2+) release in mutant G4828A. It also restored caffeine-induced Ca(2+) release in mutants I4829A and I4829T. In single-channel recordings, mutants I4829V and G4830A retained normal conductance, whereas all others had decreased unitary channel conductances ranging from 27 to 540 picosiemens. Single-channel modulation was retained in G4826A, I4829V, and G4830A, but was lost in other mutants. In contrast to wt and G4826A, I4829V, and G4830A, in which divalent metals were preferentially conducted, mutants with loss of modulation had no selectivity of divalent cations over a monovalent cation. Analysis of Gly(4822) to Asp(4831) mutants in RyR2 supports the view that this highly conserved sequence constitutes part of the ion-conducting pore of the Ca(2+) release channel and plays a key role in ryanodine and caffeine binding and activation. PMID- 11427531 TI - Evidence that inhibition of cathepsin-B contributes to the neuroprotective properties of caspase inhibitor Tyr-Val-Ala-Asp-chloromethyl ketone. AB - During the use of tetrapeptide and other proprietary caspase inhibitors in the study of neurodegeneration, we had concluded that mechanisms other than the inhibition of caspases contributed to the protective effects of certain caspase inhibitors. Here we report our studies to identify a target for and hence a mechanism by which the tetrapeptide inhibitor tyrosine-valine-alanine-aspartate chloromethyl ketone (Ac-YVAD-cmk) is able to rescue neuronal cell cultures from cell death. Ac-YVAD-cmk rescued neuronal cells from cell death in response to oxidative stress and oxygen/glucose deprivation. Affinity labeling with biotinylated YVAD-cmk demonstrated distinct binding proteins for the inhibitor in cells from the central nervous system versus Jurkat cells. Binding to the novel target protein was displaced by class-specific protease inhibitors and suggested that the target is a cysteine protease. Affinity purification and sequencing identified the target as cathepsin-B. Cathepsin-B inhibitors competed with biotinylated YVAD-cmk for the target protein. The availability of the target for binding was reduced in cells that had been rescued by unlabeled inhibitor. Cathepsin-B inhibitors rescue hippocampal slices from cell death induced by oxygen/glucose deprivation. These data provide evidence to support a role for cathepsin-B in neuronal cell death, particularly that following ischemia. PMID- 11427532 TI - p53 Modulates the exonuclease activity of Werner syndrome protein. AB - Werner syndrome (WS) is characterized by the early onset of symptoms of premature aging, cancer, and genomic instability. The molecular basis of the defects is not understood but presumably relates to the DNA helicase and exonuclease activities of the protein encoded by the WRN gene that is mutated in the disease. The attenuation of p53-mediated apoptosis in WS cells and reported physical interaction between WRN and the tumor suppressor p53 suggest that p53 and WRN functionally interact in a pathway necessary for the normal cellular response. In this study, we have demonstrated that p53 inhibits the exonuclease activity of the purified full-length recombinant WRN protein. p53 did not have an effect on a truncated amino-terminal WRN fragment that retains exonuclease activity but lacks the physical interaction domain for p53 located in the carboxyl terminus. Two naturally occurring p53 mutants found in human cancer displayed a reduced ability to inhibit WRN exonuclease activity. In cells arrested in S phase with hydroxyurea, WRN exits the nucleolus and colocalizes with p53 in the nucleoplasm. The regulation of WRN function by p53 is likely to play an important role in the maintenance of genomic integrity and prevention of cancer and other clinical symptoms associated with WS. PMID- 11427533 TI - TGIF2 interacts with histone deacetylase 1 and represses transcription. AB - TG-interacting factor (TGIF) is a transcriptional repressor, which represses transcription by binding directly to DNA or interacts with transforming growth factor beta (TGF beta)-activated Smads, thereby repressing TGF beta-responsive gene expression. Mutation of TGIF in humans causes holoprosencephaly, a severe genetic disorder affecting craniofacial development. Searching human expressed sequence tag data bases revealed the presence of clones encoding a TGIF-related protein (TGIF2), which contains two regions of high sequence identity with TGIF. Here we show that, like TGIF, TGIF2 recruits histone deacetylase, but in contrast to TGIF, is unable to interact with the corepressor CtBP. TGIF2 and TGIF have very similar DNA-binding homeodomains, and TGIF2 represses transcription when bound to DNA via a TGIF binding site. TGIF2 interacts with TGF beta-activated Smads and represses TGF beta-responsive transcription. TGIF2 appears to be a context-independent transcriptional repressor, which can perform similar functions to TGIF and may play a role in processes, which, when disrupted by mutations in TGIF, cause holoprosencephaly. PMID- 11427534 TI - A unique beta-hairpin protruding from AAA+ ATPase domain of RuvB motor protein is involved in the interaction with RuvA DNA recognition protein for branch migration of Holliday junctions. AB - The Escherichia coli RuvB protein is a motor protein that forms a complex with RuvA and promotes branch migration of Holliday junctions during homologous recombination. This study describes the characteristics of two RuvB mutants, I148T and I150T, that do not promote branch migration in the presence of RuvA. These RuvB mutants hydrolyzed ATP and bound duplex DNA with the same efficiency as wild-type RuvB, but the mutants did not form a complex with RuvA and were defective in loading onto junction DNA in a RuvA-assisted manner. A recent crystallographic study revealed that Ile(148) and Ile(150) are in a unique beta hairpin that protrudes from the AAA(+) ATPase domain of RuvB. We propose that this beta-hairpin interacts with hydrophobic residues in the mobile third domain of RuvA and that this interaction is vital for the RuvA-assisted loading of RuvB onto Holliday junction DNA. PMID- 11427535 TI - Mechanistic basis for kinetic differences between the rat alpha 1, alpha 2, and alpha 3 isoforms of the Na,K-ATPase. AB - Previous studies showed that the alpha 1, alpha 2, and alpha 3 isoforms of the catalytic subunit of the Na,K-ATPase differ in their apparent affinities for the ligands ATP, Na(+), and K(+). For the rat isoforms transfected into HeLa cells, K'(ATP) for ATP binding at its low affinity site is lower for alpha 2 and alpha 3 compared with alpha 1; relative to alpha 1 and alpha 2, alpha 3 has a higher K'(Na) and lower K'(K) (Jewell, E. A., and Lingrel, J. B (1991) J. Biol. Chem. 266, 16925--16930; Munzer, J. S., Daly, S. E., Jewell-Motz, E. A., Lingrel, J. B, and Blostein, R. (1994) J. Biol. Chem. 269, 16668--16676). The experiments described in the present study provide insight into the mechanistic basis for these differences. The results show that alpha 2 differs from alpha1 primarily by a shift in the E(1) E(2) equilibrium in favor of E(1) form(s) as evidenced by (i) a approximately 20-fold increase in IC(50) for vanadate, (ii) decreased catalytic turnover, and (iii) notable stability of Na,K-ATPase activity at acidic pH. In contrast, despite its lower K'(ATP) compared with alpha 1, the E(1) E(2) poise of alpha 3 is not shifted toward E(1). Distinct intrinsic interactions with Na(+) ions are underscored by the marked selectivity for Na(+) over Li(+) of alpha 3 compared with either alpha1 or alpha 2 and higher K'(Na) for cytoplasmic Na(+), which persists over a 100-fold range in proton concentration, independent of the presence of K(+). The kinetic analysis also suggests alpha 3-specific differences in relative rates of partial reactions, which impact this isoform's distinct apparent affinities for both Na(+) and K(+). PMID- 11427536 TI - The anaerobic ribonucleotide reductase from Lactococcus lactis. Interactions between the two proteins NrdD and NrdG. AB - Deoxyribonucleotide synthesis by anaerobic class III ribonucleotide reductases requires two proteins, NrdD and NrdG. NrdD contains catalytic and allosteric sites and, in its active form, a stable glycyl radical. This radical is generated by NrdG with its [4Fe-4S](+) cluster and S-adenosylmethionine. We now find that NrdD and NrdG from Lactobacillus lactis anaerobically form a tight alpha(2)beta(2) complex, suggesting that radical generation by NrdG and radical transfer to the specific glycine residue of NrdD occurs within the complex. Activated NrdD was separated from NrdG by anaerobic affinity chromatography on dATP-Sepharose without loss of its glycyl radical. NrdD alone then catalyzed the reduction of CTP with formate as the electron donor and ATP as the allosteric effector. The reaction required Mg(2+) and was stimulated by K(+) but not by dithiothreitol. Thus NrdD is the actual reductase, and NrdG is an activase, making class III reductases highly similar to pyruvate formate lyase and its activase and suggesting a common root for the two anaerobic enzymes during early evolution. Our results further support the contention that ribonucleotide reduction during transition from an RNA world to a DNA world started with a class III-like enzyme from which other reductases evolved when oxygen appeared on earth. PMID- 11427537 TI - Identification of genes induced in peripheral nerve after injury. Expression profiling and novel gene discovery. AB - Peripheral nerve injury results in axonal degeneration and in phenotypic changes of the surrounding Schwann cells, whose presence is critical for nerve regeneration. To identify genes induced after nerve injury in Schwann cells, we developed a strategy that included differential screening of a subtractive library enriched for cDNAs expressed in injured nerve, sequence analysis, and expression profiling. By using real time quantitative reverse transcriptase polymerase chain reaction, we found that injury-induced genes could be categorized into four temporal expression patterns. Among the clones we identified were a number that were homologous only to expressed sequence tags in the data base. These were stratified based on their expression profile, presence of identifiable sequence motifs, homologies to other proteins, and evolutionary conservation. We chose one representative gene, nin283, to analyze in detail. The nin283 gene encodes a 227-residue protein containing both a zinc finger and a RING finger motif. nin283 is highly expressed in the central nervous system, particularly in the developing cortical plate in embryos. It is also expressed in peripheral ganglia and is induced by nerve growth factor in PC12 cells. Subcellular localization analysis demonstrated that Nin283 is located in the endosome/lysosome compartment, suggesting that it may participate in ubiquitin mediated protein modification. PMID- 11427538 TI - Sphingosine 1-phosphate may be a major component of plasma lipoproteins responsible for the cytoprotective actions in human umbilical vein endothelial cells. AB - Sphingosine 1-phosphate (S1P), a novel lipid mediator, is concentrated in the fraction of lipoproteins that include high density lipoprotein (HDL) and low density lipoprotein (LDL) in human plasma. Here, we show that oxidation of LDL resulted in a marked reduction in the S1P level in association with a marked accumulation of lysophosphatidylcholine (LPC). We therefore investigated the role of the lipoprotein-associated lipids especially S1P in the lipoprotein-induced cytoprotective or cytotoxic actions in human umbilical vein endothelial cells. The viability of the cells gradually decreased in the absence of serum or growth factors in the culture medium. The addition of oxidized LDL (ox-LDL) accelerated the decrease in the cell viability. LPC and 7-ketocholesterol mimicked ox-LDL actions. On the other hand, HDL and LDL almost completely reversed the serum deprivation- or ox-LDL-induced cytotoxicity. Exogenous S1P mimicked cytoprotective actions. Moreover, the S1P-rich fraction and chromatographically purified S1P from HDL exerted cytoprotective actions, but the rest of the fractions did not. The cytoprotective actions of HDL and S1P were associated with extracellular signal-regulated kinase (ERK) activation and were almost completely inhibited by pertussis toxin and PD98059, an ERK kinase inhibitor. The HDL induced action was specifically desensitized in the S1P-pretreated cells. Taken together, these results indicate that the lipoprotein-associated S1P and the lipid receptor-mediated signal pathways may be responsible for the lipoprotein induced cytoprotective actions. Furthermore, the decrease in the S1P content, in addition to the accumulation of cytotoxic substances such as LPC, may be important for the acquisition of the cytotoxic property to ox-LDL. PMID- 11427539 TI - Dnmt3a and Dnmt3b are transcriptional repressors that exhibit unique localization properties to heterochromatin. AB - We demonstrate that the recently identified DNA methyltransferases, Dnmt3a and Dnmt3b, like DNMT1, repress transcription in a methylation-independent manner. Dnmt3a and Dnmt3b repress transcription primarily through a plant homeodomain like motif that is shared with the ATRX protein but is not present in DNMT1. Unlike DNMT1, which localizes to replication foci during S-phase in murine embryonic fibroblasts, Dnmt3a co-localizes with heterochromatin protein 1 alpha (HP1 alpha) and methyl-CpG binding proteins throughout the cell cycle to late replicating pericentromeric heterochromatin. In contrast to Dnmt3a, Dnmt3b remained diffuse in the nucleus of embryonic fibroblasts at all cell cycle stages. However, Dnmt3a and Dnmt3b co-localize to these pericentromeric heterochromatin regions in murine embryonic stem cells. This finding is important to the fact that mutations in DNMT3B are found in the developmental syndrome, ICF (immunodeficiency, centromeric heterochromatin instability, and facial anomalies), which involves extensive loss of methylation from pericentromeric regions. The localization of Dnmt3a and Dnmt3b was unaffected in Dnmt1 null embryonic stem cells, which lose the majority of methylation at pericentromeric major satellite repeats, suggesting that these enzymes are not dependent upon preexisting methylation for their targeting. DNMT1 is then positioned to reestablish transcriptionally repressive chromatin as cells replicate, while Dnmt3a and Dnmt3b may help to establish such chromatin in late S-phase and maintain this repressive heterochromatin throughout the cell cycle in a developmentally and/or cell type manner. PMID- 11427540 TI - A naturally occurring nonpolymerogenic mutant of alpha 1-antitrypsin characterized by prolonged retention in the endoplasmic reticulum. AB - The classical form of alpha 1-antitrypsin (alpha 1-AT) deficiency is associated with a mutant alpha 1-ATZ molecule that polymerizes in the endoplasmic reticulum (ER) of liver cells. A subgroup of individuals homozygous for the protease inhibitor (PI) Z allele develop chronic liver injury and are predisposed to hepatocellular carcinoma. In this study we evaluated the primary structure of alpha 1-AT in a family in which three affected members had severe liver disease associated with alpha 1-AT deficiency. We discovered that one sibling was a compound heterozygote with one PI Z allele and a second allele, the PI Z + saar allele, bearing the mutation that characterizes alpha 1-ATZ as well as the mutation that characterizes alpha 1-AT Saarbrucken (alpha 1-AT saar). The mutation in PI saar introduces a premature termination codon resulting in an alpha 1-AT protein truncated for 19 amino acids at its carboxyl terminus. Studies of a second sib with severe liver disease and other living family members did not reveal the presence of the alpha 1-AT saar mutation and therefore do not substantiate a role for this mutation in the liver disease phenotype of this family. However, studies of alpha 1-AT saar and alpha 1-ATZ + saar expressed in heterologous cells show that there is prolonged intracellular retention of these mutants even though they do not have polymerogenic properties. These results therefore have important implications for further understanding the fate of mutant alpha 1-AT molecules, the mechanism of ER retention, and the pathogenesis of liver injury in alpha 1-AT deficiency. PMID- 11427541 TI - Glycosylation increases potassium channel stability and surface expression in mammalian cells. AB - N-linked glycosylation is not required for the cell surface expression of functional Shaker potassium channels in Xenopus oocytes (Santacruz-Toloza, L., Huang, Y., John, S. A., and Papazian, D. M. (1994) Biochemistry 33, 5607-5613). We have now investigated whether glycosylation increases the stability, cell surface expression, and proper folding of Shaker protein expressed in mammalian cells. The turnover rates of wild-type protein and an unglycosylated mutant (N259Q,N263Q) were compared in pulse-chase experiments. The wild-type protein was stable, showing little degradation after 48 h. In contrast, the unglycosylated mutant was rapidly degraded (t(1/2) = approximately 18 h). Lactacystin slowed the degradation of the mutant protein, implicating cytoplasmic proteasomes in its turnover. Rapid lactacystin-sensitive degradation could be conferred on wild-type Shaker by a glycosylation inhibitor. Expression of the unglycosylated mutant on the cell surface, assessed using immunofluorescence microscopy and biotinylation, was dramatically reduced compared with wild type. Folding and assembly were analyzed by oxidizing intersubunit disulfide bonds, which provides a fortuitous hallmark of the native structure. Surprisingly, formation of disulfide-bonded adducts was quantitatively similar in the wild-type and unglycosylated mutant proteins. Our results indicate that glycosylation increases the stability and cell surface expression of Shaker protein but has little effect on acquisition of the native structure. PMID- 11427542 TI - Potential impact of the VITEK 2 system and the Advanced Expert System on the clinical laboratory of a university-based hospital. AB - A study was designed to assess the impact of the VITEK 2 automated system and the Advanced Expert System (AES) on the clinical laboratory of a typical university based hospital. A total of 259 consecutive, nonduplicate isolates of Enterobacteriaceae members, Pseudomonas aeruginosa, and Staphylococcus aureus were collected and tested by the VITEK 2 system for identification and antimicrobial susceptibility testing, and the results were analyzed by the AES. The results were also analyzed by a human expert and compared to the AES analyses. Among the 259 isolates included in this study, 245 (94.6%) were definitively identified by VITEK 2, requiring little input from laboratory staff. For 194 (74.9%) isolates, no inconsistencies between the identification of the strain and the antimicrobial susceptibility determined by VITEK 2 were detected by the AES. Thus, no input from laboratory staff was required for these strains. The AES suggested one or more corrections to results obtained with 65 strains to remove inconsistencies. The human expert thought that most of these corrections were appropriate and that some resulted from a failure of the VITEK 2 system to detect certain forms of resistance. Antimicrobial phenotypes assigned to the strains by the AES for beta-lactams, aminoglycosides, quinolones, macrolides, tetracyclines, and glycopeptides were similar to those assigned by the human expert for 95.7 to 100% of strains. These results indicate that the VITEK 2 system and AES can provide accurate information in tests for most of the clinical isolates examined and remove the need for human analysis of results for many. Certain problems were identified in the study that should be remediable with further work on the software supporting the AES. PMID- 11427543 TI - Evaluation of methods for subtyping Campylobacter jejuni during an outbreak involving a food handler. AB - In October 1998, the Centers for Disease Control and Prevention (CDC) assisted in an investigation of an outbreak of campylobacteriosis at a school in Salina, Kansas. Twenty-two isolates were submitted from the Kansas state public health laboratory to CDC, 9 associated with the outbreak and 13 epidemiologically unrelated sporadic isolates. Pulsed-field gel electrophoresis (PFGE) using SmaI and SalI was initially used to validate the epidemiologic data. We then tested the ability of other subtyping techniques to distinguish the outbreak-associated isolates from unrelated sporadic isolates. The methods employed were somatic O serotyping, PCR-restriction fragment length polymorphism (RFLP) analysis of flaA, DNA sequence analysis of 582 bp of flaA that included the short variable region (SVR), and sequencing of the entire flaA gene. PFGE was the most discriminatory technique, yielding 11 SmaI and 10 SalI restriction profiles. All outbreak isolates were indistinguishable by PFGE, somatic O serotyping, and sequencing of the 582-bp region of the flaA gene. fla typing by PCR-RFLP grouped one sporadic isolate with the outbreak strain. Analysis of the DNA sequence of a 582-bp segment of flaA produced strain groupings similar to that generated by PCR-RFLP but further differentiated two flaA PCR-RFLP types (with a 1-bp difference in the 582-bp region). Two sporadic strains were distinct by flaA PCR-RFLP but differed only by a single base substitution in the 582-bp region. The entire flaA gene was sequenced from strains differing by a single base pair in the 582-bp region, and the data revealed that additional discrimination may in some cases be obtained by sequencing outside the SVR. PFGE was superior to all other typing methods tested for strain discrimination; it was crucial for understanding the Kansas outbreak and, when SmaI was used, provided adequate discrimination between unrelated isolates. PMID- 11427544 TI - Development of molecular methods for identification of Schizophyllum commune from clinical samples. AB - In the last 50 years, to our knowledge, only 16 cases of diseases caused by Schizophyllum commune in humans have been reported. Within only 6 months, we found four isolates of this basidiomycetous fungus, obtained from patients suffering from chronic sinusitis. The cultures of the isolated fungi showed neither clamp connections nor fruiting bodies (basidiocarps), which are distinctive features for S. commune, but fast-growing cottony white mycelium only. This was harvested, and DNA was extracted. The internal transcribed spacer region of the ribosomal DNA (rDNA) was amplified with fungus-specific primers, and the PCR products were sequenced. Two strains of S. commune, collected from branches of a European hornbeam (Carpinus betulus) and a tree of heaven (Ailanthus altissima), respectively; four specimens from the herbarium of the Institute of Botany, Karl-Franzens-University Graz; and two strains from internationally known culture collections (CBS 340.81 [ATCC 44201] and CBS 405.96) were investigated in the same way. The sequence data of all strains were compared and showed homology of over 99% in this 660-bp-long fragment of rDNA. With these results, a map of restriction enzyme cutting sites and a primer set specific for S. commune were created for reliable identification of this human pathogenic fungus. PMID- 11427545 TI - Truncated hantavirus nucleocapsid proteins for serotyping Hantaan, Seoul, and Dobrava hantavirus infections. AB - Truncated recombinant nucleocapsid proteins (rNPs) of Hantaan virus (HTNV), Seoul virus (SEOV), and Dobrava virus (DOBV) were expressed by a baculovirus system. The truncated rNPs, which lacked 49 (rNP50) or 154 (rNP155) N-terminal amino acids of the NPs of HTNV, SEOV, and DOBV, were able to differentiate HTNV-, SEOV , and DOBV-specific immune sera. Recombinant NP50s retained higher reactivities than rNP155s and were proven useful for enzyme-linked immunosorbent assay (ELISA). The ELISAs based on the rNP50s of HTNV, SEOV, and DOBV successfully differentiated three groups of patient sera, previously defined by neutralization tests: 17 with HTNV infection, 12 with SEOV infection, and 20 with DOBV infection. The entire rNP of Puumala virus (PUUV) distinguished PUUV infection from the other types of hantavirus infection. Serotyping with these rNP50s can be recommended as a rapid and efficient system for hantavirus diagnosis. PMID- 11427546 TI - Genetic diversity and biochemical characteristics of Trichosporon asahii isolated from clinical specimens, houses of patients with summer-type-hypersensitivity pneumonitis, and environmental materials. AB - Trichosporon asahii, which is distributed in the environment, is the major causative agent of the opportunistic infection trichosporonosis, and it also causes summer-type hypersensitivity pneumonitis (SHP). Random amplification of polymorphic DNA analysis was used to determine the intraspecies diversity of 39 T. asahii isolates from clinical specimens, SHP patients' houses, and environmental materials. The three primers used revealed 46 polymorphic bands. A phenogram was generated by the unweighted pair-group method with arithmetic mean. Clinical isolates formed a cluster, characterized by a 90% matching coefficient, but they did not cluster with strains isolated from SHP patients' houses or environmental sources. In addition, the biochemical characteristics of 86 strains from three sources were examined with 31 compounds using an ID32C kit, and a phenogram was constructed. The phenogram consisted of three major clusters. Cluster I included most of the clinical SHP isolates, and cluster II included most of the environmental isolates. Cluster III contained only one strain. A remarkable difference was found in the abilities of the strains belonging to clusters I and II to utilize six compounds. These results suggest that the genetic diversity and biochemical characteristics of T. asahii seem to be related to the source of the isolate. We also found a specific DNA fragment for the clinical isolates and strains isolated from SHP patients' houses. PMID- 11427547 TI - Helicobacter winghamensis sp. nov., a novel Helicobacter sp. isolated from patients with gastroenteritis. AB - From 1997 to 1999 seven isolates of Campylobacter-like organisms from five patients that were exhibiting symptoms of gastroenteritis, including fever, stomach malaise, and diarrhea, were investigated. The organisms were isolated from stool samples and found to exhibit a diverse colony morphology; hence multiple isolates were submitted from one of the patients. All isolates were found to be identical. The organisms were catalase, urease, alkaline phosphatase, and nitrate negative but oxidase and indoxyl acetate positive. They grew at 37 degrees C but not at 42 degrees C, and three of the isolates from two different patients were sensitive to nalidixic acid and cephalothin. Full 16S rRNA sequence analysis not only grouped these organisms within the Helicobacter genus but also differentiated them from previously identified Helicobacter species. The closest relative by phylogenetic analysis was Helicobacter sp. flexispira taxon 1. Electron microscopy showed that these isolates had one or two bipolar flagella; however, the periplasmic fibers, a characteristic of the known Helicobacter sp. flexispira taxa, were not observed. The present isolates also lacked a flagellar sheath, a trait shared with four other Helicobacter spp., H. canadensis, H. mesocricetorum, H. pullorum, and H. rodentium. On the basis of the unique phenotypic properties of these isolates and 16S rRNA sequence analysis, we propose the classification of a new Helicobacter species, Helicobacter winghamensis sp. nov. PMID- 11427548 TI - Combinatorial use of antibodies to secreted mycobacterial proteins in a host immune system-independent test for tuberculosis. AB - Laboratory diagnosis of tuberculosis is often difficult. Immunodetection of circulating Mycobacterium tuberculosis proteins shed during active infection would not depend on an intact host immune response and could take advantage of the speed and low costs afforded by antibody-based assays. We previously showed that patients with active tuberculosis had increased levels of circulating antigen 85 (Ag85) proteins independent of their tuberculin skin test status (S. I. Bentley-Hibbert, X. Quan, T. Newman, K. Huygen, and H. P. Godfrey, Infect. Immun. 67:581-588, 1999). To extend these observations to a Mycobacterium bovis BCG-vaccinated population and to another secreted mycobacterial protein, Ag85 and PstS-1 (protein antigen B, p38 antigen) were quantified in sera from 97 Chilean tuberculosis patients and healthy controls (many of whom had received BCG as children) using dot immunobinding, mouse monoclonal anti-BCG Ag85 complex antibody, and chicken antipeptide antibodies reactive with M. tuberculosis Ag85B and PstS-1. The latter antibodies had been raised to peptide-derived immunogens expressed on a novel proprietary protein carrier in Escherichia coli. Median serum Ag85 levels measured by using either anti-Ag85 antibody were significantly higher in patients with active tuberculosis than in healthy controls (P, <0.001 to 0.01); the median serum PstS-1 levels were similar in patients and controls. The sensitivity of significantly elevated circulating Ag85 levels in patients with pulmonary tuberculosis measured by anti-Ag85 complex or anti-Ag85B antibodies was 60 and 55%, respectively, but increased to 77% when results obtained with both anti-Ag85 antibodies were considered jointly (P < 0.02). The corresponding specificities for individual and joint consideration were 95, 85, and 80%, respectively. These results indicate that elevated Ag85 levels can be detected in patients with active tuberculosis even after BCG vaccination and suggest that combinatorial use of antibodies directed at different epitopes of this protein could provide a viable strategy for developing new host immune response-independent diagnostic tests for tuberculosis. PMID- 11427549 TI - rpoB sequence analysis of cultured Tropheryma whippelii. AB - Until recently no isolate of Tropheryma whippelii was available, and therefore genetic studies were limited to those based on PCR amplification of conserved genes. In this study we determined the nucleotide sequence of rpoB (encoding the beta-subunit of RNA polymerase) from a cultured strain of T. whippelii using degenerate consensus PCR and genome walking. The T. whippelii rpoB consists of 3,657 bp with a 50.4% GC content and encodes 1,218 amino acids with a calculated molecular mass of 138 kDa. Comparison of T. whippelii RpoB with other eubacterial RpoB proteins indicated sequence similarity ranging from 57.19 (Mycoplasma pneumoniae) to 74.63% (Mycobacterium tuberculosis). Phylogenetic analysis of T. whippelii based on comparison of its RpoB sequence with sequences available for other bacteria was consistent with that previously derived from the 16S ribosomal DNA (rDNA) sequence, indicating that it belongs to the actinomyces clade. The sequence comparison allowed the design of a primer pair, TwrpoB.F and TwrpoB.R, specific for T. whippelii rpoB. When incorporated into a PCR, this primer pair allowed the detection of T. whippelii rpoB in three of three 16S rDNA PCR positive biopsy specimens and zero of seven negative controls. rpoB could therefore be targeted in PCR-mediated detection and identification of this emerging bacterial species. This approach has previously been shown useful for the identification of related mycobacteria. This study underscores that a method involving isolation and then propagation of emerging bacteria is a useful way to quickly achieve extensive molecular knowledge of these pathogens. PMID- 11427550 TI - Identification of two different 14-alpha sterol demethylase-related genes (cyp51A and cyp51B) in Aspergillus fumigatus and other Aspergillus species. AB - Two cyp51-related genes (cyp51A and cyp51B) encoding 14-alpha sterol demethylase like enzymes were identified in the opportunistic human pathogen Aspergillus fumigatus. PCR amplification using degenerate oligonucleotides based on conserved areas of cytochrome P450 demethylases of other filamentous fungi and yeasts allowed the cloning and sequencing of two different homologue genes in A. fumigatus. Southern analysis confirmed that both genes hybridized to distinct genomic loci and that both are represented as single copies in the genome. Comparison of the deduced Cyp51A and Cyp51B proteins with the CYP51 proteins from Penicillium italicum, Aspergillus nidulans, Erysiphe graminis, Uncinula necator, Botrytis cinerea, Ustilago maydis, Cryptococcus neoformans, Candida albicans, Saccharomyces cerevisiae, Candida tropicalis, and Candida glabrata showed that the percentages of identity of the amino acid sequences (range, 40 to 70%) were high enough to consider Cyp51A and Cyp51B to be members of the fungal CYP51 family. Fragments from both genes were also cloned from other Aspergillus spp. (A. flavus, A. nidulans, and A. terreus). Phylogenetic analysis showed that, at least in the most pathogenic species of Aspergillus, there are two fungal CYP51 proteins. This is the first report of the existence of two homologue genes coding for 14-alpha sterol demethylase in the fungal kingdom. This finding could provide insights into the azole resistance mechanisms operating in fungi. The primers used here may be useful molecular tools for facilitating the cloning of novel 14 alpha sterol demethylase genes in other filamentous fungi. PMID- 11427551 TI - Evaluation of current methods for detection of staphylococci with reduced susceptibility to glycopeptides. AB - The sensitivity and specificity of seven methods (agar dilution, broth microdilution, Etest at 0.5 and 2.0 McFarland (McF) inocula, two agar screening methods, and population studies [PS]) were evaluated in a double-blind study involving 284 methicillin-resistant Staphylococcus aureus (MRSA) strains and 45 Staphylococcus strains with reduced susceptibilities to vancomycin (SRSV). The results were compared to the population analysis profile-area under the curve ratio method (PAP-AUC ratio compared to that of Mu3) as described by Wootton et al. The agar screening method using brain heart infusion agar (6 microg of vancomycin per ml) gave a sensitivity of 22% and a specificity of 97%. A similar method using Mueller-Hinton agar (5 microg of vancomycin per ml) gave a sensitivity of 20% and a specificity of 99%. The PS method detected 34 false positives (12%) and gave a sensitivity of 71% and a specificity of 88%. Etest using 0.5 and 2.0 McF inocula gave sensitivities and specificities of 82 and 93% and of 96 and 97%, respectively. The best Etest interpretative criteria for the 2.0 McF inoculum was > or =8 mg of vancomycin per liter and > or =8 microg teicoplanin per ml or > or =12 microg of teicoplanin per ml. The direct colony suspension inoculum for this method was found to be equally accurate in detecting (hetero-)glycopeptide-intermediate S. aureus compared to the overnight broth inoculum preparation method. Agar dilution and broth microdilution using the NCCLS breakpoint criteria for vancomycin gave sensitivities and specificities of 20 and 100% and of 11 and 100%, respectively. Using the Etest with a 2.0 McF inoculum, six different media were assessed against a selection of SRSV (n = 48) and MRSA (n = 12). Brain heart infusion agar yielded the highest sensitivity and specificity values: 88 and 88%, respectively. PMID- 11427552 TI - Diagnosis of Oropouche virus infection using a recombinant nucleocapsid protein based enzyme immunoassay. AB - Oropouche (ORO) virus is an emerging infectious agent that has caused numerous outbreaks of an acute febrile (dengue-like) illness among humans in Brazil, Peru, and Panama. Diagnosis of ORO virus infection is based mainly on serology. Two different antigens, hamster serum antigen (HSA) and Vero cell lysate antigen (VCLA), are currently used in enzyme immunoassays (EIAs) in Brazil and Peru, respectively, to investigate the epidemiology of ORO virus infection. Both antigens involve use of infectious virus, and for this reason their use is restricted. Consequently, the frequency and distribution of ORO virus infection are largely unexplored in other countries of South America. This report describes the use of a bacterially expressed recombinant nucleocapsid (rN) protein of ORO virus in EIAs for the diagnosis of ORO virus infection. The data revealed that the purified rN protein is comparable to the authentic viral N protein in its antigenic characteristics and is highly sensitive and specific in EIAs. Among 183 serum samples tested, a high degree of concordance was found between rN protein based EIA and HSA- and VCLA-based EIAs for the detection of both ORO virus specific immunoglobulin M (IgM) and IgG antibodies. The high sensitivity, specificity, and safety of the rN protein-based EIA make it a useful diagnostic technique that can be widely used to detect ORO virus infection in South America. PMID- 11427553 TI - Comparison of variable number tandem repeat and IS6110-restriction fragment length polymorphism analyses for discrimination of high- and low-copy-number IS6110 Mycobacterium tuberculosis isolates. AB - The present study was designed to evaluate the use of variable number tandem repeat (VNTR) and IS6110-restriction fragment length polymorphism (RFLP) analyses in combination as a two-step strategy for discrimination (as measured by the Hunter-Gaston Discrimination Index [HGDI]) of both high- and low-copy-number IS6110 Mycobacterium tuberculosis isolates compared to IS6110-RFLP alone with an unselected collection of isolates. Individually, IS6110-RFLP fingerprinting produced six clusters that accounted for 69% of the low-copy-number IS6110 isolates (five clusters) and 5% of the high-copy-number IS6110 isolates (one cluster). A total of 39% of all the isolates were clustered (HGDI = 0.97). VNTR analysis generated a total of 35 different VNTR allele profile sets from 93 isolates (HGDI = 0.938). Combining IS6110-RFLP analysis with VNTR analysis reduced the overall percentage of clustered isolates to 29% (HGDI = 0.988) and discriminated a further 27% of low-copy-number isolates that would have been clustered by IS6110-RFLP alone. The use of VNTR analysis as an initial typing strategy facilitates further analysis by IS6110-RFLP, and more importantly, VNTR analysis subdivides some IS6110-RFLP-defined clusters containing low- and single copy IS6110 isolates. PMID- 11427554 TI - Antifungal susceptibility testing of fluconazole by flow cytometry correlates with clinical outcome. AB - Susceptibility testing of fungi by flow cytometry (also called fluorescence activated cell sorting [FACS]) using vital staining with FUN-1 showed a good correlation with the standard M27-A procedure for assessing MICs. In this study we determined MICs for blood culture isolates from patients with candidemia by NCCLS M27-A and FACS methods and correlated the clinical outcome of these patients with in vitro antifungal resistance test results. A total of 24 patients with candidemia for whom one or more blood cultures were positive for a Candida sp. were included. Susceptibility testing was performed by NCCLS M27-A and FACS methods. The correlation of MICs (NCCLS M27-A and FACS) and clinical outcome was calculated. In 83% of the cases, the MICs of fluconazole determined by FACS were within 1 dilution of the MICs determined by the NCCLS M27-A method. For proposed susceptibility breakpoints, there was 100% agreement between the M27-A and FACS methods. In the FACS assay, a fluconazole MIC of <1 microg/ml was associated with cure (P < 0.001) whereas an MIC of > or =1 microg/ml was associated with death (P < 0.001). The M27-A-derived fluconazole MICs did not correlate with outcome (P = 1 and P = 0.133). PMID- 11427555 TI - Clinical relevance of the babA2 genotype of Helicobacter pylori in Japanese clinical isolates. AB - Genotypic variation of Helicobacter pylori is speculated to associate with different clinical outcomes. In Western countries, the gene encoding blood group antigen-binding adhesin (BabA), babA2, is of high clinical relevance and is a useful marker to identify patients who are at higher risk for peptic ulceration and gastric adenocarcinoma, as are vacA and cagA. We investigated the presence of babA2 and cagA in 179 Japanese clinical isolates by PCR and Southern blot analysis and looked for correlations with various clinical outcomes (nonulcer dyspepsia, duodenal ulcers, gastric ulcers gastric adenocarcinoma, and mucosa associated lymphoid tissue lymphoma). The prevalence of the babA2 genotype was 84.9% and that of the cagA genotype was 96.1%. There was no correlation between the babA2 and cagA genotypes, and there was no association between the babA2 or cagA status and clinical outcome. These results indicate that babA2 status is not of high clinical relevance in Japan and that Japanese strains are different from those infecting Western populations. PMID- 11427556 TI - Serodiagnosis of human granulocytic ehrlichiosis by using novel combinations of immunoreactive recombinant proteins. AB - A panel of seven recombinant antigens, derived from Ehrlichia phagocytophila (the agent of human granulocytic ehrlichiosis), was evaluated by class-specific enzyme linked immunosorbent assays (ELISAs) for utility in the diagnosis of the infection. Fourteen genomic fragments, obtained by serologic expression screening, contained open reading frames (ORFs) encoding 16 immunodominant antigens. Eleven of these antigens were members of the major surface protein (MSP) multigene family. Alignment of their predicted protein sequences revealed a pattern of conserved sequences, which contained short direct repeats, flanking a variable region. In addition, two genomic clones contained two and three MSP ORFs, respectively, indicating that these genes are clustered in tandem copies. The implications for this pattern of both genomic and protein arrangements in antigenic variations of MSPs and in their utilities in a diagnostic assay are discussed. In addition to two MSP recombinant antigens (rHGE-1 and -3) and a fusion protein of these antigens (rErf-1), five further recombinants were evaluated by ELISA. Two of these antigens (rHGE-14 and -15) were novel, while a third (rHGE-2), with no known function, has been described. The final two recombinant antigens (rHGE-9 and -17) represent overlapping segments of the ankyrin gene (ank). The addition of rHGE-9 ELISA data resulted in the detection of 78% (21 of 27) of acute-phase sera. When serologic data for all recombinants are combined, 96.2% (26 of 27) of convalescent-phase patient serum samples and 85.2% (23 of 27) of acute-phase patient serum samples are detected, indicating the potential of these antigens for use in the development of a rapid serologic assay for the detection of E. phagocytophila infection. PMID- 11427557 TI - Comparison of cultivation and PCR-hybridization for detection of Salmonella in porcine fecal and water samples. AB - A total of 150 fecal and water samples from four swine farms were tested for the presence of Salmonella enterica using different enrichment techniques as follows: (i) 92 fecal samples from nursery and farrowing barns at three swine farms were preenriched overnight in tryptic soy broth (TSB) at 37 degrees C followed by overnight enrichment in Rappaport-Vassiliadis 10 broth (RV10) at 42 degrees C; (ii) 24 water samples from the third farm were preenriched overnight in 3MC broth at 37 degrees C followed by overnight enrichment in RV10 at 42 degrees C; and (iii) 34 fecal samples from a fourth farm, a finishing farm, were enriched overnight in RV10 at 42 degrees C with no additional enrichment. Following each of the enrichment techniques, samples were subcultured onto modified semisolid Rappaport-Vassiliadis (MSRV) agar prior to transfer to Hektoen Enteric agar plates for the recovery of viable Salmonella bacteria. Presumptive Salmonella isolates were biochemically and serologically confirmed. For the PCR detection of Salmonella, a 1-ml portion was removed from each sample after the first overnight enrichment and the DNA was extracted using a Sepharose CL-6B spin column. Amplicons (457 bp) derived from primers to the invA and invE genes were confirmed as Salmonella specific on ethidium bromide-stained agarose gels by Southern hybridization with a 20-mer oligonucleotide probe specific for the Salmonella invA gene. Neither the standard microbiological method nor the molecular method detected all of the 65 samples that tested positive by both methods or either method alone. Salmonella bacteria were detected by both cultivation and PCR hybridization in 68% (17 of 25) of the positive samples that were preenriched in TSB, in 73% (11 of 15) of the positive samples preenriched in 3MC broth, and in 24% (6 of 25) of the positive samples enriched in RV10. Agreement between Salmonella detection using cultivation with preenrichment and detection by PCR was 76% using the kappa statistic. However, agreement between Salmonella detection using cultivation without preenrichment and detection by PCR was about 6%; the PCR assay detected 80% (20 of 25) of the 25 positive samples, while Salmonella bacteria were recovered from only 44% (11 of 25) by cultivation. Our results indicate that the PCR-hybridization approach is equivalent to or better than cultivation for detecting Salmonella in swine feces or water samples from swine farms when using the medium combinations evaluated in this study. PMID- 11427558 TI - Serological expression cloning and immunological evaluation of MTB48, a novel Mycobacterium tuberculosis antigen. AB - Improved diagnostics are needed for the detection of Mycobacterium tuberculosis, especially for patients with smear-negative disease. To address this problem, we have screened M. tuberculosis (H37Rv and Erdman strains) genomic expression libraries with pooled sera from patients with extrapulmonary disease and with sera from patients with elevated reactivity with M. tuberculosis lysate. Both serum pools were reactive with clones expressing a recombinant protein referred to here as MTB48. The genomic sequence of the resulting clones was identical to that of the M. tuberculosis H37Rv isolate and showed 99% identity to the Mycobacterium bovis and M. bovis BCG isolate sequences. The genomic location of this sequence is 826 bp upstream of a region containing the esat-6 gene that is deleted in the M. bovis BCG isolate. The mtb48 1,380-bp open reading frame encodes a predicted 47.6-kDa polypeptide with no known function. Southern and Western blot analyses indicate that this sequence is present in a single copy and is conserved in the M. tuberculosis and M. bovis isolates tested but not in other mycobacterial species tested, including Mycobacterium leprae and Mycobacterium avium. In addition, the native protein was detected in the cytoplasm, as was a processed form that was also shed into the medium during culture. Serological analysis of recombinant MTB48 and the M. tuberculosis 38-kDa antigen with a panel of patient and control sera indicates that the inclusion of recombinant MTB48 in a prototype serodiagnostic test increases assay sensitivity for M. tuberculosis infection when it is combined with other known immunodominant antigens, such as the 38-kDa antigen. PMID- 11427559 TI - Recombinant major antigenic protein 2 of Ehrlichia canis: a potential diagnostic tool. AB - The major antigenic protein 2 (MAP2) of Ehrlichia canis was cloned and expressed. The recombinant protein was characterized and tested in an enzyme-linked immunosorbent assay (ELISA) format for potential application in the serodiagnosis of canine monocytic ehrlichiosis. The recombinant protein, which contained a C terminal polyhistidine tag, had a molecular mass of approximately 26 kDa. The antigen was clearly identified by Western immunoblotting using antihistidine antibody and immune serum from an experimentally infected dog. The recombinant MAP2 (rMAP2) was tested in an ELISA format using 141 serum samples from E. canis immunofluorescent antibody (IFA)-positive and IFA-negative dogs. Fifty-five of the serum samples were from dogs experimentally or naturally infected with E. canis and were previously demonstrated to contain antibodies reactive with E. canis by indirect immunofluorescence assays. The remaining 86 samples, 33 of which were from dogs infected with microorganisms other than E. canis, were seronegative. All of the samples from experimentally infected animals and 36 of the 37 samples from naturally infected animals were found to contain antibodies against rMAP2 of E. canis in the ELISA. Only 3 of 53 IFA-negative samples tested positive on the rMAP2 ELISA. There was 100% agreement among IFA-positive samples from experimentally infected animals, 97.3% agreement among IFA-positive samples from naturally infected animals, and 94.3% agreement among IFA-negative samples, resulting in a 97.2% overall agreement between the two assays. These data suggest that rMAP2 of E. canis could be used as a recombinant test antigen for the serodiagnosis of canine monocytic ehrlichiosis. PMID- 11427560 TI - Genetic heterogeneity of Borrelia burgdorferi sensu lato in the southern United States based on restriction fragment length polymorphism and sequence analysis. AB - Fifty-six strains of Borrelia burgdorferi sensu lato, isolated from ticks and vertebrate animals in Missouri, South Carolina, Georgia, Florida, and Texas, were identified and characterized by PCR-restriction fragment length polymorphism (RFLP) analysis of rrf (5S)-rrl (23S) intergenic spacer amplicons. A total of 241 to 258 bp of intergenic spacers between tandemly duplicated rrf (5S) and rrl (23S) was amplified by PCR. MseI and DraI restriction fragment polymorphisms were used to analyze these strains. PCR-RFLP analysis results indicated that the strains represented at least three genospecies and 10 different restriction patterns. Most of the strains isolated from the tick Ixodes dentatus in Missouri and Georgia belonged to the genospecies Borrelia andersonii. Excluding the I. dentatus strains, most southern strains, isolated from the ticks Ixodes scapularis and Ixodes affinis, the cotton rat (Sigmodon hispidus), and cotton mouse (Peromyscus gossypinus) in Georgia and Florida, belonged to Borrelia burgdorferi sensu stricto. Seven strains, isolated from Ixodes minor, the wood rat (Neotoma floridana), the cotton rat, and the cotton mouse in South Carolina and Florida, belonged to Borrelia bissettii. Two strains, MI-8 from Florida and TXW-1 from Texas, exhibited MseI and DraI restriction patterns different from those of previously reported genospecies. Eight Missouri tick strains (MOK-3a group) had MseI patterns similar to that of B. andersonii reference strain 21038 but had a DraI restriction site in the spacer. Strain SCGT-8a had DraI restriction patterns identical to that of strain 25015 (B. bissettii) but differed from strain 25015 in its MseI restriction pattern. Strain AI-1 had the same DraI pattern as other southern strains in the B. bissettii genospecies but had a distinct MseI profile. The taxonomic status of these atypical strains needs to be further evaluated. To clarify the taxonomic positions of these atypical Borrelia strains, the complete sequences of rrf-rrl intergenic spacers from 20 southeastern and Missouri strains were determined. The evolutionary and phylogenetic relationships of these strains were compared with those of the described genospecies in the B. burgdorferi sensu lato species complex. The 20 strains clustered into five separate lineages on the basis of sequence analysis. MI-8 and TXW-1 appeared to belong to two different undescribed genospecies, although TXW-1 was closely related to Borrelia garinii. The MOK-3a group separated into a distinct deep branch in the B. andersonii lineage. PCR-RFLP analysis results and the results of sequence analyses of the rrf-rrl intergenic spacer confirm that greater genetic heterogeneity exists among B. burgdorferi sensu lato strains isolated from the southern United States than among strains isolated from the northern United States. The B. andersonii genospecies and its MOK-3a subgroup are associated with the I. dentatus-cottontail rabbit enzootic cycle, but I. scapularis was also found to harbor a strain of this genospecies. Strains that appear to be B. bissettii in our study were isolated from I. minor and the cotton mouse, cotton rat, and wood rat. The B. burgdorferi sensu stricto strains from the south are genetically and phenotypically similar to the B31 reference strain. PMID- 11427561 TI - Evaluation of a modified sanitary napkin as a sample self-collection device for the detection of genital chlamydial infection in women. AB - A modified sanitary napkin was compared with endocervical swab and urine specimens for the detection of urogenital Chlamydia trachomatis infection. Endocervical swabs and/or first-catch urine were collected from 510 women at medical or community settings in Quebec City. Participants were also asked to wear a modified sanitary napkin (Ezy-Detek) during 4 consecutive hours and to bring it back to the clinic or mail it to the laboratory. Endocervical and urine specimens were tested using the Cobas Amplicor CT/NG assay (Roche Diagnostic Systems) according to the manufacturer's instructions, as were specimens collected with the napkin after adequate preparation. If the PCR test result was positive on the endocervical sample or on any two samples, a woman was considered to be infected. PCR testing results on paired samples were identical for 493 (96.6%) of 510 women. According to the definition given above, 58 (11.3%; 95% confidence interval [CI], 8.7 to 14.5%) women were infected with C. trachomatis. The sensitivity and specificity of PCR testing on modified sanitary napkin specimens were, respectively, 93.1% (54 of 58; 95% CI, 83.3 to 98.1%) and 98.9% (447 of 452; 95% CI, 97.4 to 99.6%) compared to 81.0% (47 of 58; 95% CI, 68.6 to 90.1%) and 100% (451 of 451; 95% CI, 99.2 to 100%) for urine specimens. The positive and negative predictive values were, respectively, 91.5% (54 of 59) and 99.1% (447 of 451) for the sanitary napkin specimens compared to 100% (47 of 47) and 97.6% (451 of 462) for urine samples. These results suggest that a modified sanitary napkin represents an effective noninvasive device for self-collection of specimens to detect urogenital C. trachomatis infection. PMID- 11427562 TI - Standardization of antifungal susceptibility variables for a semiautomated methodology. AB - Recently, the methodology that will serve as a basis of the standard for antifungal susceptibility testing of fermentative yeasts of the European Committee on Antibiotic Susceptibility Testing has been described. This procedure employs a spectrophotometric method for both inoculum adjustment and endpoint determination. However, the utilization of a spectrophotometer requires studies for standardization. The present work analyzes the following parameters: (i) accuracy of inoculum preparation, (ii) correlation between optical density and CFU per milliliter, (iii) influence of the wavelength on the endpoint determination, and (iv) influence of the dimethyl sulfoxide concentration on the growth kinetics. The main results can be summarized as follows: (i) inoculum preparation following the methodology recommended by the National Committee for Clinical Laboratory Standards is an exact procedure; (ii) the relationship between optical density and CFU per milliliter is linear (coefficient of determination, r(2) = 0.84); (iii) MICs obtained by means of spectrophotometric readings at different wavelengths are identical (for amphotericin B, an intraclass correlation coefficient of 0.98 was obtained; for fluconazole, the intraclass correlation coefficient was 1); and (iv) a 2% concentration of dimethyl sulfoxide produces a significantly slower and lower growth curve of Candida spp. than other concentrations. PMID- 11427563 TI - Fourth-generation enzyme-linked immunosorbent assay for the simultaneous detection of human immunodeficiency virus antigen and antibody. AB - The VIDAS HIV DUO Ultra, a fourth-generation immunoassay under development for the simultaneous detection of human immunodeficiency virus type 1 (HIV-1) p24 antigen and antibodies to HIV-1 and HIV-2, was evaluated. The enzyme-linked fluorescence immunoassay, performed on the automated VIDAS instrument, is claimed to detect early and established HIV infection. The assay was challenged with a total of 2,847 samples that included 74 members of 10 seroconversion panels, 9 p24 antigen-only-reactive members of a panel of group M clades, 503 consecutively collected samples from individuals seeking care in the University of Maryland Medical System, 1,010 samples from U.S. blood donors, 1,141 samples from patients in a high-incidence population in Trinidad, 83 samples from a clinic for sexually transmitted diseases in the Bahamas, 10 confirmed HIV-1 group O samples, and 16 confirmed HIV-2 samples from the Cote d'Ivoire. Reference tests were U.S. Food and Drug Administration-licensed HIV antibody screening, p24 antigen tests, HIV confirmatory assays, and the Roche Diagnostics Amplicor HIV-1 Monitor. The VIDAS HIV DUO Ultra demonstrated 100% sensitivity and 99.5% specificity overall, with a 99.7% specificity in low-risk individuals. The analytical sensitivity, as assessed by seroconversion panels and p24 antigen in samples, was equivalent to the sensitivity of the reference assays used to characterize these panels. The VIDAS HIV DUO Ultra is accurate, offers potential advantages over conventional HIV testing for time and cost savings, has walk-away capability, and correctly identifies both early and established HIV infections. PMID- 11427564 TI - Interaction between Mycoplasma hyopneumoniae and swine influenza virus. AB - An experimental respiratory model was used to investigate the interaction between Mycoplasma hyopneumoniae and swine influenza virus (SIV) in the induction of pneumonia in susceptible swine. Previous studies demonstrated that M. hyopneumoniae, which produces a chronic bronchopneumonia in swine, potentiates a viral pneumonia induced by the porcine reproductive and respiratory syndrome virus (PRRSV). In this study, pigs were inoculated with M. hyopneumoniae 21 days prior to inoculation with SIV. Clinical disease as characterized by the severity of cough and fever was evaluated daily. Percentages of lung tissue with visual lesions and microscopic lesions were assessed upon necropsy at 3, 7, 14, and 21 days following SIV inoculation. Clinical observations revealed that pigs infected with both SIV and M. hyopneumoniae coughed significantly more than pigs inoculated with a single agent. Macroscopic pneumonia on necropsy at days 3 and 7 was greatest in both SIV-infected groups, with minimal levels of pneumonia in the M. hyopneumoniae-only-infected pigs. At 14 days post-SIV inoculation, pneumonia was significantly more severe in pigs infected with both pathogens. However, by 21 days postinoculation, the level of pneumonia in the dual-infected pigs was similar to that of the M. hyopneumoniae-only-infected group, and the pneumonia in the pigs inoculated with only SIV was nearly resolved. Microscopically, there was no apparent increase in the severity of pneumonia in pigs infected with both agents compared to that of single-agent-challenged pigs. The results of this study found that while pigs infected with both agents exhibited more severe clinical disease, the relationship between the two pathogens lacked the profound potentiation found with dual infection with M. hyopneumoniae and PRRSV. These findings demonstrate that the relationship between mycoplasmas and viruses varies with the individual agent. PMID- 11427565 TI - Identification of rifampin-resistant Mycobacterium tuberculosis strains by hybridization, PCR, and ligase detection reaction on oligonucleotide microchips. AB - Three new molecular approaches were developed to identify drug-resistant strains of Mycobacterium tuberculosis using biochips with oligonucleotides immobilized in polyacrylamide gel pads. These approaches are significantly faster than traditional bacteriological methods. All three approaches-hybridization, PCR, and ligase detection reaction--were designed to analyze an 81-bp fragment of the gene rpoB encoding the beta-subunit of RNA polymerase, where most known mutations of rifampin resistance are located. The call set for hybridization analysis consisted of 42 immobilized oligonucleotides and enabled us to identify 30 mutant variants of the rpoB gene within 24 h. These variants are found in 95% of all mutants whose rifampin resistance is caused by mutations in the 81-bp fragment. Using the second approach, allele-specific on-chip PCR, it was possible to directly identify mutations in clinical samples within 1.5 h. The third approach, on-chip ligase detection reaction, was sensitive enough to reveal rifampin resistant strains in a model mixture containing 1% of resistant and 99% of susceptible bacteria. This level of sensitivity is comparable to that from the determination of M. tuberculosis drug resistance by using standard bacteriological tests. PMID- 11427566 TI - Development of a PCR assay for identification of staphylococci at genus and species levels. AB - We have developed a PCR-based assay which allows the detection of staphylococci at the genus level by targeting the tuf gene, which encodes the elongation factor Tu. Degenerate PCR primers derived from consensus regions of several tuf genes were used to amplify a target region of 884 bp from 11 representative staphylococcal species. Subsequently, the entire nucleotide sequence of these amplicons was determined. The analysis of a multiple alignment of these sequences revealed regions conserved among staphylococci but distinct from those of other gram-positive bacteria genetically related to staphylococci. PCR primers complementary to these regions could amplify specifically and efficiently a DNA fragment of 370 bp for all of 27 different staphylococcal species tested. There was no amplification with genomic DNA prepared from 53 nonstaphylococcal species tested to verify the specificity of the assay (20 gram positive and 33 gram negative). Furthermore, this assay amplified efficiently all 27 American Type Culture Collection (ATCC) staphylococcal reference strains as well as 307 clinical isolates of staphylococci from the Quebec City region. Analysis of the multiple sequence alignment for the 884-bp fragment for the 11 staphylococcal species as well as comparison of the sequences for the 370-bp amplicon from five unrelated ATCC and clinical strains for each of the species S. aureus, S. epidermidis, S. haemolyticus, S. hominis, and S. saprophyticus demonstrated sufficient interspecies polymorphism to generate genus- and species-specific capture probes. This sequence information allowed the development of Staphylococcus-specific and species-specific (targeting S. aureus, S. epidermidis, S. haemolyticus, S. hominis, or S. saprophyticus) capture probes hybridizing to the 370-bp amplicon. In conclusion, this PCR assay is suitable for detection of staphylococci at both genus and species levels. PMID- 11427567 TI - Genomic heterogeneity and O-antigenic diversity of Campylobacter upsaliensis and Campylobacter helveticus strains isolated from dogs and cats in Germany. AB - A serotyping scheme based on heat-stable surface antigens was established for 101 Campylobacter upsaliensis and 10 Campylobacter helveticus strains isolated from 261 dogs and 46 cats of different ages originating from two geographically distinct regions in Germany. The prevalence of C. upsaliensis varied between 27.8% in juvenile dogs (<12 months of age) and 55.4% in adult dogs (P < 0.05). Of the cats, 19.6% harbored C. upsaliensis, whereas 21.7% carried C. helveticus. Of the C. upsaliensis isolates from both host species, 93.1% belonged to five different serogroups, two of them being prevalent at rates of 47.5 and 27.7%, with different frequencies in both regions. Six (54.6%) of the C. helveticus isolates also belonged to serotypes found among C. upsaliensis strains, whereas five (45.4%) possessed an O antigen unique for C. helveticus. In contrast, a considerable degree of genomic diversity of the isolates was assessed by macrorestriction analyses with the endonucleases SmaI and XhoI, using pulsed field gel electrophoresis as well as enterobacterial repetitive intergenic consensus sequence PCR (ERIC PCR). Restriction with SmaI pointed towards the existence of clonal groups associated to some extent with serotypes, while restriction with XhoI disintegrated these groups to smaller noncoherent subgroups. Analysis of ERIC PCR profiles did not exhibit any associations with serotypes. In conclusion these data demonstrate the genomic heterogeneity among C. upsaliensis strains and indicate that the combination of SmaI restriction with serotyping is a useful tool to investigate the expansion of clonal groups of C. upsaliensis. PMID- 11427568 TI - Pasteurella multocida subsp. multocida and P. multocida subsp. septica differentiation by PCR fingerprinting and alpha-glucosidase activity. AB - Pasteurella multocida is composed of three subspecies that are often differentiated by fermentation of sorbitol and dulcitol. We studied 35 dulcitol negative P. multocida isolates from infected dog and cat bite wounds, 16 of which yielded weak and/or conflicting fermentation reactions in Andrades sorbitol, thus making it difficult to distinguish between the two dulcitol-negative subspecies of P. multocida, i.e., P. multocida subsp. multocida and P. multocida subsp. septica. All isolates and two control strains were further analyzed using a PCR fingerprinting technique with a single primer (M13 core) and assessed for alpha glucosidase (alpha-Glu) activity. Although the PCR fingerprint patterns and alpha Glu activity did not correlate well with the sorbitol fermentation reactions, they did correlate well with each other. All strains identified as P. multocida subsp. septica were positive for alpha-Glu activity and exhibited the group I PCR fingerprint profile. All strains categorized as P. multocida subsp. multocida displayed either the group II or group III PCR fingerprint profile; 9 of 11 of these isolates were alpha-Glu negative. These data suggest that both PCR fingerprinting and alpha-Glu activity provide reliable means for differentiating P. multocida subsp. multocida from P. multocida subsp. septica, particularly in strains that produce weak and/or discrepant sorbitol fermentation reactions. PMID- 11427569 TI - Nomenclature of major antimicrobial-resistant clones of Streptococcus pneumoniae defined by the pneumococcal molecular epidemiology network. AB - The emergence of disease caused by penicillin-resistant and multidrug-resistant pneumococci has become a global concern, necessitating the identification of the epidemiological spread of such strains. The Pneumococcal Molecular Epidemiology Network was established in 1997 under the auspices of the International Union of Microbiological Societies with the aim of characterizing, standardizing, naming, and classifying antimicrobial agent-resistant pneumococcal clones. Here we describe the nomenclature for 16 pneumococcal clones that have contributed to the increase in antimicrobial resistance worldwide. Guidelines for the recognition of these clones using molecular typing procedures (pulsed-field gel electrophoresis, BOX-PCR, and multilocus sequence typing) are presented, as are the penicillin binding profiles and macrolide resistance determinants for the 16 clones. This network can serve as a prototype for the collaboration of scientists in identifying clones of important human pathogens and as a model for the development of other networks. PMID- 11427570 TI - Suitability of a rapid immunochromatographic test for detection of antibodies to human immunodeficiency virus in Ghana, West Africa. AB - In West African countries such as Ghana, efficient human immunodeficiency virus (HIV) testing is a priority in the fight against AIDS. A new immunochromatographic rapid test, Determine HIV-1/2 (Abbott Diagnostics, North Chicago, Ill.), that detects antibodies against HIV type 1 (HIV-1) and/or HIV-2 was evaluated using Ghanaian blood samples. Two hundred four serum and/or plasma specimens were tested. HIV screening was done by a particle agglutination test and confirmed by a Western blot (WB) test as the "gold standard." The results revealed 125 HIV-seropositive AIDS patients, 75 HIV-seronegative healthy individuals, and 4 individuals for whom the HIV-1 result was indeterminate. The results obtained by the Determine HIV-1/2 assay and Diagnostic HIV SPOT (Genelabs), which is currently widely used in many districts in Ghana, were compared with those of the WB test, excluding the four HIV-1-indeterminate samples. The sensitivity of the Determine HIV-1/2 assay was 100%, compared with 98.0% for the HIV SPOT assay. The specificity was 100% for both tests. Determine HIV-1/2 is a single-step assay and was found to be rapid and easy to perform without any special equipment. It was highly sensitive and specific. The kit can be applied without electricity and water supplies, making it suitable for the detection of HIV antibodies especially in the rural areas of Ghana, West Africa. PMID- 11427571 TI - O-antigen diversity among Acinetobacter baumannii strains from the Czech Republic and Northwestern Europe, as determined by lipopolysaccharide-specific monoclonal antibodies. AB - O-antigen-specific monoclonal antibodies (MAbs) are currently being generated to develop an O-serotyping scheme for the genus Acinetobacter and to provide potent tools to study the diversity of O-antigens among Acinetobacter strains. In this report, Acinetobacter baumannii strains from the Czech Republic and from two clonal groups identified in Northwestern Europe (termed clones I and II) were investigated for their reactivity with a panel of O-antigen-specific MAbs generated against Acinetobacter strains from various species. The bacteria were characterized for their ribotype, biotype, and antibiotic susceptibility and the presence of the 8.7-kb plasmid pAN1. By using the combination of these typing profiles, the Czech strains could be classified into four previously defined groups (A. Nemec, L. Janda, O. Melter, and L. Dijkshoorn, J. Med. Microbiol. 48:287-296, 1999): two relatively homogeneous groups of multiresistant strains (termed groups A and B), a heterogeneous group of other multiresistant strains, and a group of susceptible strains. O-antigen reactivity was observed primarily with MAbs generated against Acinetobacter calcoaceticus and Acinetobacter baumannii strains. A comparison of reaction patterns confirmed the previously hypothesized clonal relationship between group A and clone I strains, which are also similar in other properties. The results show that there is limited O antigen variability among strains with similar geno- and phenotypic characteristics and are suggestive of a high prevalence of certain A. baumannii serotypes in the clinical environment. It is also shown that O-antigen-specific MAbs are useful for the follow-up of strains causing outbreaks in hospitals. PMID- 11427572 TI - Performance of the chromogenic medium CHROMagar Staph Aureus and the Staphychrom coagulase test in the detection and identification of Staphylococcus aureus in clinical specimens. AB - CHROMagar Staph aureus (CSAM) (CHROMagar Microbiology, Paris, France) is a new chromogenic medium designed to enable detection of colonies of Staphylococcus aureus by their pink color. A total of 775 specimens were cultured in parallel on CHROMagar Staph aureus and conventional media. Among the 267 S. aureus strains recovered on at least one medium, 263 were isolated on CSAM medium (sensitivity, 98.5%), and 245 (sensitivity, 91.8%) were isolated on conventional media. The specificity of presumptive identification of S. aureus on the basis of pink colony color on CSAM medium was 97% (493 of 508). This specificity increased to 100% when coagulase detection with the Staphychrom coagulase test was added and to 98.8% when S. aureus surface components were detected by agglutination in the Pastorex Staph Plus test. Susceptibility testing of 67 S. aureus strains, performed in parallel on pink CSAM colonies and on colonies grown on blood agar, gave similar results. Thus, rapid and accurate recognition and identification of S. aureus isolates were achieved with CSAM as the primary isolation medium, followed by the staphylocoagulase Staphychrom test. Antimicrobial susceptibility testing (disk-diffusion method or ATB STAPH System) can be performed directly on pink CSAM colonies. PMID- 11427573 TI - Development of a rapid and sensitive test for identification of major pathogens in bovine mastitis by PCR. AB - Bovine mastitis is the most important source of loss for the dairy industry. A rapid and specific test for the detection of the main pathogens of bovine mastitis is not actually available. Molecular probes reacting in PCR with bacterial DNA from bovine milk, providing direct and rapid detection of Escherichia coli, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus parauberis, and Streptococcus uberis, have been developed. Two sets of specific primers were designed for each of these microorganisms and appeared to discriminate close phylogenic bacterial species (e.g., S. agalactiae and S. dysgalactiae). In addition, two sets of universal primers were designed to react as positive controls with all major pathogens of bovine mastitis. The sensitivities of the test using S. aureus DNA extracted from milk with and without a pre-PCR enzymatic lysis step of bacterial cells were compared. The detection limit of the assay was 3.125 x 10(2) CFU/ml of milk when S. aureus DNA was extracted with the pre-PCR enzymatic step compared to 5 x 10(3) CFU/ml of milk in the absence of the pre-PCR enzymatic step. This latter threshold of sensitivity is still compatible with its use as an efficient tool of diagnosis in bovine mastitis, allowing the elimination of expensive reagents. The two PCR tests avoid cumbersome and lengthy cultivation steps, can be performed within hours, and are sensitive, specific, and reliable for the direct detection in milk of the six most prevalent bacteria causing bovine mastitis. PMID- 11427574 TI - Diagnosis of bacteriuria by detection of volatile organic compounds in urine using an automated headspace analyzer with multiple conducting polymer sensors. AB - The Osmetech Microbial Analyzer (OMA) is an automated headspace analyzer fitted with a novel detector system consisting of an array of polymer sensors, each of which responds to different volatile organic compounds. The system can be used for screening clinical urine specimens for significant bacteriuria by sampling urine headspace and subjecting the output of the multiple-detector response to principal component analysis. The OMA readily distinguished artificially infected urine samples from sterile controls. The OMA was then used to analyze 534 unselected clinical urine specimens, of which 21.5% had significant bacteriuria (containing >10(5) CFU of bacteria/ml). The sensitivity and specificity of the OMA compared with conventional culture were 83.5 and 87.6%, respectively. The OMA is a promising automated system for the rapid routine screening of urine specimens, and further clinical trials are in progress. PMID- 11427575 TI - Detection of RTX toxin gene in Vibrio cholerae by PCR. AB - A PCR that amplifies a recently discovered Vibrio cholerae RTX (repeat in toxin) toxin gene was developed. Among 166 clinical and environmental isolates of V. cholerae causing epidemics and sporadic cases of cholera in various parts of the world, all were found to be toxigenic by both PCR and HEp-2 cell cytotoxicity assay. Standard strains of the classical biotype containing a deletion within the gene cluster exhibited negative results by both assays. This is the first rapid genotyping method for differentiation of V. cholerae O1 classical biotype strains from El Tor biotype strains as well as strains of other non-O1 serogroups including serogroup O139. The PCR assay that was developed also specifically detects RTX toxin genes in V. cholerae, as clinical isolates of Vibrio parahaemolyticus, diarrheagenic Escherichia coli, Aeromonas species, and Plesiomonas species were all negative by the RTX toxin-specific PCR as well as the HEp-2 cytotoxicity assay. These findings highlight the characteristics of the RTX toxins in V. cholerae. Their role in the pathogenicity of the bacterium requires further investigation. PMID- 11427576 TI - Fluorogenic PCR-based quantitative detection of a murine pathogen, Helicobacter hepaticus. AB - Helicobacter hepaticus infection in mice is being used as an animal model for elucidating the pathogenesis of gastrointestinal and biliary diseases in humans. H. hepaticus, which forms a spreading film on selective agar, is not amenable to routine quantitative counts of organisms in tissues using a CFU method. In this study, a fluorogenic PCR-based assay was developed to quantitatively detect H. hepaticus in mouse ceca and feces using the ABI Prism 7700 sequence detection system. A pair of primers and a probe for this assay were generated from the H. hepaticus cdtB gene (encoding subunit B of the H. hepaticus cytolethal distending toxin). Using this assay, the sensitivity for detection of H. hepaticus chromosomal DNA prepared from pure culture was 20 fg, which is equivalent to approximately 14 copies of the H. hepaticus genome based on an estimated genome size of approximately 1.3 Mb determined by pulsed-field gel electrophoresis. H. hepaticus present in feces and cecal samples from H. hepaticus-infected mice was readily quantified. The selected PCR primers and probe did not generate fluorescent signals from eight other helicobacters (H. canis, H. cineadi, H. felis, H. mustelae, H. nemestrinae, H. pullorum, H. pylori, and H. rodentium). A fluorescent signal was detected from 20 ng of H. bilis DNA but with much lower sensitivity (10(6)-fold) than from H. hepaticus DNA. Therefore, this assay can be used with high sensitivity and specificity to quantify H. hepaticus in experimentally infected mouse models as well as in naturally infected mice. PMID- 11427577 TI - Identification and expression of a 50-kilodalton surface antigen of Babesia gibsoni and evaluation of its diagnostic potential in an enzyme-linked immunosorbent assay. AB - A cDNA expression library prepared from Babesia gibsoni merozoite mRNA was screened with B. gibsoni-infected dog serum. cDNA encoding a 50-kDa protein was cloned and designated the P50 gene. The complete nucleotide sequence of the P50 gene was 1,922 bp. Computer analysis suggested that the sequence of the P50 gene contained an open reading frame of 1,401 bp with a coding capacity of approximately 50 kDa. The complete genomic nucleotide sequence of the P50 gene has been analyzed and shown to contain a single intron of 37 bp. Southern blotting analysis indicated that the P50 gene was present at a single copy in the B. gibsoni genome. The native P50 protein of B. gibsoni with a molecular mass of 50 kDa was identified by Western blotting with anti-recombinant P50 mouse serum. Confocal laser microscopic analysis showed that the P50 protein was located on the surface of B. gibsoni merozoites. The recombinant P50 protein expressed by baculovirus in insect cells was used as the antigen in an enzyme-linked immunosorbent assay (ELISA). The ELISA was able to differentiate between B. gibsoni-infected dog serum and B. canis-infected dog serum or noninfected dog serum. Furthermore, the antibody response against the recombinant P50 protein was maintained until the chronic stage of infection in dogs experimentally infected with B. gibsoni was developed. These results demonstrate that the recombinant P50 protein might be a useful diagnostic reagent for detection of antibodies to B. gibsoni in dogs. PMID- 11427578 TI - Detection of rifampin-resistant Mycobacterium tuberculosis in sputa by nested PCR linked single-strand conformation polymorphism and DNA sequencing. AB - Either PCR-mediated single strand conformation polymorphism (SSCP) analysis or DNA sequencing of rpoB DNA (157 bp) can be used as a rapid screening method for the detection of mutations related to the rifampin resistance of Mycobacterium tuberculosis. However, due to the nonspecific amplification of rpoB DNA from nontuberculous mycobacteria these methods cannot be directly applied to clinical specimens such as sputa. We developed a nested PCR method that can specifically amplify the rpoB DNA of M. tuberculosis on the basis of rpoB DNA sequences of 44 mycobacteria. Nested PCR-linked SSCP analysis and the DNA sequencing method were applied directly in order to detect M. tuberculosis and determine its rifampin susceptibility in 56 sputa. The results obtained by nested PCR-SSCP and DNA sequencing were concordant with those of conventional drug susceptibility testing and DNA sequencing performed with culture isolates. PMID- 11427579 TI - Direct detection of Legionella species from bronchoalveolar lavage and open lung biopsy specimens: comparison of LightCycler PCR, in situ hybridization, direct fluorescence antigen detection, and culture. AB - We developed a rapid thermocycling, real-time detection (also known as real-time PCR) method for the detection of Legionella species directly from clinical specimens. This method uses the LightCycler (Roche Molecular Biochemicals, Indianapolis, Ind.) and requires approximately 1 to 2 h to perform. Both a Legionella genus PCR assay and Legionella pneumophila species-specific PCR assay were designed. A total of 43 archived specimens from 35 patients were evaluated, including 19 bronchoalveolar lavage (BAL) specimens and 24 formalin-fixed, paraffin-embedded open lung biopsy specimens. Twenty-five of the specimens were culture-positive for Legionella (9 BAL specimens and 16 tissue specimens). BAL specimens were tested by LightCycler PCR (LC-PCR) methods and by a direct fluorescent antibody (DFA) assay, which detects L. pneumophila serogroups 1 to 6 and several other Legionella species. Tissue sections were tested by the two LC PCR methods, by DFA, by an in situ hybridization (ISH) assay, specifically designed to detect L. pneumophila, and by Warthin-Starry (WS) staining. The results were compared to the "gold standard" method of bacterial culture. With BAL specimens the following assays yielded the indicated sensitivities and specificities, respectively: Legionella genus detection by Legionella genus LC PCR, 100 and 100%; Legionella genus detection by DFA assay, 33 and 100%; and L. pneumophila detection by L. pneumophila species-specific LC-PCR, 100 and 100%. With open lung biopsy specimens the following assays yielded the indicated sensitivities and specificities, respectively: Legionella genus detection by LC PCR 68.8 and 100%; Legionella genus detection by DFA assay, 44 and 100%; Legionella genus detection by WS staining, 63 and 100%; L. pneumophila species specific detection by LC-PCR, 17 and 100%; and L. pneumophila species-specific detection by ISH, 100 and 100%. The analytical sensitivity of both LC-PCR assays was <10 CFU/reaction. LC-PCR is a reliable method for the direct detection of Legionella species from BAL specimens. The Legionella genus LC-PCR assay could be performed initially; if positive, L. pneumophila species-specific LC-PCR could then be performed (if species differentiation is desired). The speed with which the LC-PCR procedure can be performed offers significant advantages over both culture-based methods and conventional PCR techniques. In contrast, for the methods evaluated, culture was the best for detecting multiple Legionella species in lung tissue. WS staining, Legionella genus LC-PCR, and L. pneumophila species specific ISH were useful as rapid tests with lung tissue. PMID- 11427580 TI - Specific detection of Pasteurella multocida in chickens with fowl cholera and in pig lung tissues using fluorescent rRNA in situ hybridization. AB - A Pasteurella multocida species-specific oligonucleotide probe, pmhyb449, targeting 16S rRNA was designed and evaluated by whole-cell hybridization against 22 selected reference strains in animal tissues. It differentiated P. multocida from other bacterial species of the families Pasteurellaceae and Enterobacteriaceae and also from divergent species of the order Cytophagales (except biovar 2 strains of Pasteurella avium and Pasteurella canis, which have high 16S rRNA similarity to P. multocida). The potential of the probe for specific identification and differentiation of P. multocida was further detected in formalin-fixed paraffin-embedded lung tissues from experimental fowl cholera in chickens and infections in pigs. In chicken lung tissues P. multocida cells were detected singly, in pairs, as microcolonies, and as massive colonies within air capillaries (septa and lumen), parabronchial septa, and blood vessels (wall and lumen). In pig lung, postmortem-injected P. multocida was detected in the alveoli (lumen and wall), and in both animals the bacterial cells were seen in the bronchi. The results showed that with the oligonucleotide probe pmhyb449, fluorescent in situ hybridization is a suitable and fast method for specific detection of P. multocida in histological formalin-fixed tissues. The test was replicable and reproducible and is recommended as a supplementary test for diagnosis and as a tool in pathogenesis studies of fowl cholera and respiratory tract infections in pigs due to P. multocida. PMID- 11427581 TI - Molecular identification of a Dietzia maris hip prosthesis infection isolate. AB - Dietzia maris, an environmental actinomycete, has been implicated only once in human disease. We herein report the first D. maris isolate from a bone biopsy specimen in a patient hospitalized for a total hip prosthesis replacement. Cell wall fatty acid analysis and 16S ribosomal DNA gene sequencing were utilized to achieve its definite identification. This case report illustrates the usefulness of such methods for the accurate identification of actinomycetes. PMID- 11427582 TI - Practical strategies for detecting and confirming vancomycin-intermediate Staphylococcus aureus: a tertiary-care hospital laboratory's experience. AB - The clinical microbiology laboratory plays a critical role in the detection of Staphylococcus aureus with decreased susceptibility to vancomycin. Staff education and rapid laboratory response are of utmost importance. We report on our laboratory's experience and provide recommendations for the identification and confirmation of vancomycin-intermediate S. aureus. PMID- 11427583 TI - Rapid diagnosis of Histoplasma capsulatum endocarditis using the AccuProbe on an excised valve. AB - Histoplasma capsulatum is an infrequent but serious cause of endocarditis. The definitive diagnosis requires culture, which may require a long incubation. We demonstrated the ability of the Histoplasma capsulatum AccuProbe to accurately identify this organism when applied directly on an excised valve that contained abundant yeast forms consistent with H. capsulatum. PMID- 11427584 TI - Mycolic acid index susceptibility method for Mycobacterium tuberculosis. AB - A rapid drug susceptibility test to measure the susceptibility of Mycobacterium tuberculosis to isoniazid (INH) and rifampin (RIF) using clinical isolates and a newly defined mycolic acid index (MAI) was evaluated. A total of 200 clinical isolates of M. tuberculosis were tested for susceptibility or resistance to INH and RIF by the MAI susceptibility and indirect-proportion methods. Overall, there was agreement between the two methods for 398 (99.5%) of the 400 total tests. Specifically, the sensitivity of the MAI susceptibility method for INH and RIF was 97.6 and 100%, respectively. The specificity and positive predictive value were 100% for both drugs, and the negative predictive value for INH and RIF was 98.3 and 100%, respectively. In conclusion, the MAI susceptibility method described here can be used for rapid drug susceptibility testing of M. tuberculosis clinical isolates within 5 days after clinical isolates are incubated in the presence or absence of an antituberculosis drug. PMID- 11427585 TI - Antimicrobial susceptibilities of Helicobacter pylori isolates under microaerophilic atmospheres established by two different methods. AB - The MICs of clarithromycin, amoxicillin, and metronidazole for 150 Helicobacter pylori isolates were determined using the AnaeroPack system and were compared with those determined using a microaerophilic incubator. The MICs of clarithromycin, amoxicillin, and metronidazole determined under both microaerophilic atmospheres were mostly within one twofold dilution for 146 (97.3%), 150 (100%), and 149 (99.3%) of the isolates, respectively. PMID- 11427586 TI - Clarithromycin-susceptible and -resistant Helicobacter pylori isolates with identical randomly amplified polymorphic DNA-PCR genotypes cultured from single gastric biopsy specimens prior to antibiotic therapy. AB - Of the Helicobacter pylori populations from 976 patients, six contained clarithromycin-resistant as well as -susceptible colonies. In each heterogeneous H. pylori population, resistant H. pylori colonies harbored identical 23S ribosomal DNA (rDNA) mutations associated with clarithromycin resistance, while the susceptible H. pylori colonies all had wild-type 23S rDNA. The resistant and susceptible colonies of each of the heterogeneous H. pylori populations had identical randomly amplified polymorphic DNA-PCR genotypes. In conclusion, evaluation of antimicrobial susceptibility can be misinterpreted if only a single colony from the primary H. pylori population is used to test for clarithromycin susceptibility. PMID- 11427587 TI - Resistance-associated mutations in the human immunodeficiency virus type 1 subtype c protease gene from treated and untreated patients in the United Kingdom. AB - This work reports the variability of human immunodeficiency virus type 1 (HIV-1) protease from treated and untreated patients infected with HIV-1 subtype C in the United Kingdom. The most common primary mutation observed in treated patients was L90M. D30N, M46I, V82A/F, and I84V were seen rarely. M36I and I93L mutations were observed in nearly all samples from both treated and untreated patients and so cannot be considered as resistance-associated mutations in this subtype. PMID- 11427588 TI - Use of a reverse dot blot procedure to identify the presence of multiple serovars in Chlamydia trachomatis urogenital infection. AB - Epidemiologic research requires identification of Chlamydia trachomatis serovars and detection of mixed infection. Antibody-based serotyping is unworkable when specimens are urine or vaginal swabs. We developed a reverse dot blot (RDB) to screen for multiple serotypes in these specimens. RDB yielded the predicted results on all artificially mixed samples and on seven of eight clinically mixed samples. PMID- 11427589 TI - Improved detection of methicillin-resistant Staphylococcus aureus using phenyl mannitol broth containing aztreonam and ceftizoxime. AB - We tested a phenyl mannitol broth containing ceftizoxime and aztreonam (PHMB(+)) for detection of methicillin-resistant Staphylococcus aureus (MRSA) with reference MRSA strains and, subsequently, with clinical samples (n = 1,098). All reference MRSA strains induced color change in PHMB(+) after 24 to 72 h of incubation. In a clinical setting, 40 MRSA strains were detected with PHMB(+), compared with only 23 detected with a routine method. Thus, this selective broth significantly (P < 0.001) improved the rate of MRSA detection. PMID- 11427590 TI - Sensitive detection of Borrelia burgdorferi sensu lato DNA and differentiation of Borrelia species by LightCycler PCR. AB - In order to differentiate species within the Borrelia burgdorferi sensu lato complex, LightCyler PCR and melting-curve analysis of the amplicons of two genes with intraspecies variability, the p66 gene and the recA gene, were performed. It was demonstrated that nested LightCycler PCR amplification of p66 is more sensitive in the detection of borrelia DNA than amplification of the recA gene. B. burgdorferi sensu stricto could be differentiated from Borrelia garinii and Borrelia afzelii by melting-curve analysis of the p66 gene amplicon. B. garinii could be differentiated from B. afzelii and B. burgdorferi sensu stricto by melting-curve analysis of the recA gene amplicon. Therefore, the PCRs complement each other in subtyping different Borrelia species, and combined LightCycler PCR and melting-curve analysis of both target genes is a rapid method to distinguish the three species of B. burgdorferi sensu lato. PMID- 11427591 TI - Pulmonary infection caused by an unusual, slowly growing nontuberculous Mycobacterium. AB - Mycobacterium triplex, a recently described slowly growing nontuberculous mycobacterium, was isolated from a Finnish patient with pulmonary mycobacteriosis. The disease was successfully treated with antimycobacterial drugs. The strain isolated, which was similar to the type strain but differed slightly from the species description, was regarded as a variant of M. triplex sensu stricto. According to present knowledge this variant of the species has never been isolated before. PMID- 11427592 TI - Evidence of different Enterocytozoon bieneusi genotypes in patients with and without human immunodeficiency virus infection. AB - We classified 100 Enterocytozoon bieneusi isolates into five genotypes by a PCR restriction fragment length polymorphism method. Type I strains were encountered only in human immunodeficiency virus (HIV)-infected patients, whereas type II strains were more frequently found in non-HIV-infected patients (75 versus 10%, respectively; P < 10(-4)), suggesting differences in the epidemiology of E. bieneusi among these patients. PMID- 11427593 TI - Use of real-time quantitative PCR to detect Chlamydophila felis infection. AB - A real-time PCR assay was developed to detect and quantify Chlamydophila felis infection of cats. The assay uses a molecular beacon to specifically identify the major outer membrane protein gene, is highly reproducible, and is able to detect fewer than 10 genomic copies. PMID- 11427594 TI - Increasing multidrug resistance in Helicobacter pylori strains isolated from children and adults in Mexico. AB - The susceptibilities to three antimicrobials of 195 Helicobacter pylori strains isolated from Mexican patients is reported; 80% of the strains were resistant to metronidazole, 24% were resistant to clarithromycin, and 18% presented a transient resistance to amoxicillin. Resistance to two or more antimicrobials increased significantly from 1995 to 1997. PMID- 11427595 TI - Efficacy of new low-cost filtration device for recovering Schistosoma haematobium eggs from urine. AB - A new, inexpensive filtration device for the diagnosis of urinary schistosomiasis was tested against the commonly used Millipore device. The experimental protocol was performed with 25 urine samples known to be positive for Schistosoma haematobium. The results suggest that the new device is as effective as the Millipore device for the diagnosis of urinary schistosomiasis. Its low cost will be attractive to schistosomiasis control programs. PMID- 11427596 TI - Comparison of restriction fragment length polymorphism, microsatellite length polymorphism, and random amplification of polymorphic DNA analyses for fingerprinting Aspergillus fumigatus isolates. AB - Aspergillus fumigatus fingerprints generated by random amplification of polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP) upon hybridization with repeated DNA sequences, and PCR detection of microsatellite length polymorphism (MLP) were compared among 67 isolates. In contrast to RAPD, RFLP and MLP gave discriminating and significantly concordant genotyping results. PMID- 11427597 TI - Multicenter evaluation of a pathogenic mycobacterium screening probe. AB - The introduction of nucleic acid amplification assays into the clinical laboratory has reduced the time needed to diagnose diseases caused by members of the Mycobacterium tuberculosis complex (MTBC). However, several mycobacterial species other than those of the MTBC are known to cause disease, especially in immunocompromised individuals. A screening assay has been developed for the detection of the major pathogenic mycobacterial species. The assay utilizes pan genus primers to amplify mycobacterial DNA and a screening probe (KY493) that detects all major pathogenic mycobacteria. A multicenter European study was conducted to assess the performance of the screening probe in the clinical laboratory. The screening probe was evaluated against individual probes specific for M. tuberculosis, M. avium, and M. intracellulare, a genus-specific probe with broader species coverage, and culture. The screening probe had a sensitivity equivalent to that of the species-specific probes; all specimens positive with any of the species-specific probes were also positive with the screening probes. Compared to culture, the sensitivity of the screening probe was 89% (154 of 173) for all culture-positive specimens tested. This value was 89.6% for the genus specific probe. The screening probe was more specific than the genus-specific probe. Specificity was 93.9% (661 of 704) compared to culture results alone. The comparable specificity value for the genus-specific probe was 84.8%. When clinical data were taken into consideration, the sensitivity of the screening assay was similar to that of culture (81% versus 76.2%) but the positive predictive value of the test was lower (76.2% versus 100% for culture). However, the screening probe was more sensitive than smear and may be a useful tool in the rapid diagnosis of mycobacterial disease. PMID- 11427598 TI - Heminested multiplex reverse transcription-PCR for detection and differentiation of Norwalk-like virus genogroups 1 and 2 in fecal samples. AB - The present study describes a heminested multiplex reverse transcription (RT)-PCR assay which enables simultaneous detection and differentiation of Norwalk-like virus (NLV) genogroups from clinical fecal samples without the need to perform sequencing or hybridization. The assay developed was able to detect concentrations of fewer than 100 viral particles per 5 microl of clarified fecal extract and could differentiate the two genogroups with a specificity of 100%. Although the multiplex RT-PCR assay failed to detect NLV in about 3% of the fecal samples which were NLV positive by electron microscopy (EM), the assay was approximately six times more sensitive than EM for NLV detection. PMID- 11427599 TI - Comparison of serogroup W-135 meningococci isolated in Sweden during a 23-year period and those associated with a recent hajj pilgrimage. AB - An outbreak of serogroup W-135 meningococcal disease was reported among pilgrims returning from the annual hajj (pilgrimage to Mecca) in mid-March 2000. Molecular characterization was used to investigate the similarity of the hajj-associated W 135 strains with those isolated in Sweden during a 23-year period (1978 to 2000). The same hajj-associated genosubtype, genosubtype P1.5,2,36b, has been documented in Sweden since 1979, while pulsed-field gel electrophoresis and the sulfadiazine resistance of the W-135 isolates indicated that the outbreak was probably due to a new clone of W-135 meningococci. PMID- 11427600 TI - Characterization of multidrug-resistant Salmonella enterica serovar Typhimurium isolated in Japan. AB - A total of 221 isolates of multidrug-resistant Salmonella enterica serovar Typhimurium in Japan were characterized in the present study. The results revealed that clonal serovar Typhimurium definitive phage type 104 strains prevailed and that these strains had drug resistance patterns, integron types, and pulsed-field gel electrophoresis patterns similar to those predominant among isolates in Western countries. PMID- 11427601 TI - Correlations between molecular subtyping and serotyping of Listeria monocytogenes. AB - To define relationships between Listeria monocytogenes genetic lineages, ribotypes, and serotypes, 235 L. monocytogenes isolates were characterized by serotyping and automated EcoRI ribotyping. Genetic lineage predicted the following serovar clusters: lineage I, comprising serotypes 1/2b, 3b, 3c, and 4b; lineage II, comprising serotypes 1/2a, 1/2c, and 3a; and lineage III, comprising serotypes 4a and 4c. Some EcoRI ribotypes contained multiple serotypes; a subset of these isolates was further differentiated with PvuII ribotyping. Of the 12 resultant EcoRI-PvuII combination types, only 4 contained multiple serotypes, demonstrating the potential of ribotyping for serotype prediction. PMID- 11427602 TI - Novel fluorescent broth microdilution method for fluconazole susceptibility testing of Candida albicans. AB - A comparative evaluation of the reference National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution method with a novel fluorescent carboxyfluorescein diacetate (CFDA)-modified microdilution method for the susceptibility testing of fluconazole was conducted with 68 Candida strains, including 53 Candida albicans, 5 Candida tropicalis, 5 Candida glabrata, and 5 Candida parapsilosis strains. We found trailing endpoints and discordant fluconazole MICs of < 8 microg/ml at 24 h and of > or =64 microg/ml at 48 h for 12 of the C. albicans strains. These strains satisfy the definition of the low high MIC phenotype. All 12 low-high phenotype strains were correctly shown to be susceptible at 48 h with the CFDA-modified microdilution method. For the 41 non low-high phenotype C. albicans strains, the CFDA-modified microdilution method yielded 97.6% (40 of 41 strains) agreement within +/-1 dilution at 24 h compared with the reference method and 92.7% (38 of 41 strains) agreement within +/-1 dilution at 48 h compared with the reference method. The five strains each from C. tropicalis, C. glabrata, and C. parapsilosis that were tested showed 100% agreement within +/-2 dilutions for the two methods being evaluated. PMID- 11427603 TI - Basis of the superiority of cefoperazone amphotericin teicoplanin for isolating Campylobacter upsaliensis from stools. AB - The optimum method for isolating Campylobacter upsaliensis from stools has not been clearly defined. In a preliminary study, cefoperazone amphotericin teicoplanin (CAT) selective medium isolated six C. upsaliensis strains which were not detected using modified cefoperazone charcoal deoxycholate (mCCDA). In order to identify the factors that underlie the superiority of CAT over mCCDA for isolating C. upsaliensis, we examined the effect of incubation time and antibiotic content of culture media on the growth of C. upsaliensis isolates using semiquantitative methods. The recovery of a subgroup of C. upsaliensis isolates from seeded stool specimens was also evaluated. Differences in growth of C. upsaliensis on CAT and mCCDA were modest and were not explained by the antibiotic profiles of the two media. Recovery of C. upsaliensis from spiked human feces on CAT was superior to that on mCCDA at lower concentrations of organisms (10(3) CFU/ml). We conclude that although CAT is more suitable than mCCDA for the isolation of C. upsaliensis from stools, the superiority of CAT for detecting this organism is not accounted for by the antibiotic composition of the medium. PMID- 11427604 TI - Pneumocystis carinii carriage among cystic fibrosis patients, as detected by nested PCR. AB - A total of 137 sputa from 95 consecutive cystic fibrosis (CF) patients undergoing routine bacteriological surveillance were analyzed for Pneumocystis carinii colonization using nested PCR. Seven of 95 patients (7.4%) were PCR positive, suggesting that P. carinii carriage may exist among CF patients due to their underlying pulmonary disease. PMID- 11427605 TI - Fluoroquinolone resistance is a poor surrogate marker for type II topoisomerase mutations in clinical isolates of Streptococcus pneumoniae. AB - The association between fluoroquinolone susceptibility and DNA mutations coding for amino acid substitutions in the quinolone resistance-determining region was assessed with 44 clinical isolates of Streptococcus pneumoniae. Twenty-three strains bore at least one amino acid substitution. Only seven strains with mutations were suggested by diminished susceptibility to ciprofloxacin (MIC, > or =2 microg/ml). PMID- 11427606 TI - Molecular detection of human calicivirus in young children hospitalized with acute gastroenteritis in Melbourne, Australia, during 1999. AB - Reverse transcription-PCR and sequence analysis identified calciviruses in 32 of 60 stool specimens (negative for other enteric pathogens) obtained from children admitted to our hospital with acute gastroenteritis. The overall annual incidence rate for calcivirus was 9% (32 of 354 children). Molecular analysis identified 30 "Norwalk-like virus" genogroup II (predominantly Lordsdale cluster) and 2 "Sapporo-like virus" strains. PMID- 11427607 TI - Lack of usefulness of carbon utilization tests for identification of Mycobacterium mucogenicum. AB - Carbon utilization tests have proven to be useful for the identification of some species of rapidly growing mycobacteria and have been described as one of the few tests useful for the differentiation of Mycobacterium mucogenicum from other rapid growers. We have found the carbon utilization tests to be unreliable for the identification of patient isolates of this species. In this study, using 28 isolates of rapidly growing mycobacteria, we examined several variables which might have an effect on results of citrate, inositol, and mannitol utilization: inoculum concentration, incubation temperature, and medium manufacturer. None of these variables affected results obtained for most species of rapid growers or for ATCC strains of M. mucogenicum. Results for patient isolates of M. mucogenicum were found to be inconsistent regardless of the methodology employed and resulted in an ambiguous identification of these isolates or an incorrect identification as Mycobacterium chelonae. Molecular or cell wall analysis may be the best technique to employ for accurate identification of M. mucogenicum. PMID- 11427608 TI - Comparative evaluation of penicillin, ampicillin, and imipenem MICs and susceptibility breakpoints for vancomycin-susceptible and vancomycin-resistant Enterococcus faecalis and Enterococcus faecium. AB - Although imipenem has in vitro activity against Enterococcus faecalis and Food and Drug Administration-approved indications for treatment of infections caused by this microorganism, there are no NCCLS guidelines for susceptibility testing of imipenem versus enterococci. Therefore, the in vitro activities of penicillin, ampicillin, imipenem, and vancomycin against 201 blood isolates of E. faecalis and 24 blood isolates of Enterococcus faecium were compared. The susceptibility of isolates to penicillin or ampicillin accurately predicted the in vitro activity of imipenem. Since the susceptibility of enterococci to imipenem can be predicted by the results obtained by testing of penicillin or ampicillin, testing of imipenem by clinical laboratories probably is not necessary. PMID- 11427609 TI - Effects of two different growth media on the postantifungal effect induced by polyenes on Candida species. AB - There are no data on the effects of different growth media on polyene-induced postantifungal effect (PAFE) in Candida species. Hence, the nystatin- and amphotericin B-induced PAFEs in six Candida species (26 isolates) grown in Sabouraud's dextrose broth (SAB) and RPMI broth were evaluated, following limited exposure to the MICs of the two polyenes, using an automated turbidometric method. For nystatin, PAFE varied between 1.88 and 4.87 h in SAB and 0.66 and 6.89 h in RPMI, and for amphotericin B, the equivalent values were 3.13 to 10.98 h in SAB and 0.97 to 7.01 h in RPMI. These highly significant (P < 0.001) variations in the PAFE with both drugs, noted with most Candida strains grown in different media, call for standardization of intralaboratory methodology in measuring this parameter in order to obtain universally comparable data. PMID- 11427610 TI - Limited level of accuracy provided by available rapid diagnosis tests for malaria enhances the need for PCR-based reference laboratories. AB - The rise of imported malaria cases and the high fatality rate in Europe make the search for new and easy diagnostic methods necessary. Rapid diagnosis tests (RDTs) are, in part, developed to cover the lack of diagnosis experience. Unfortunately, our data suggest that the accuracy of RDTs is insufficient and could increase the number of incorrect malaria diagnoses. PMID- 11427611 TI - Utility of 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenyl-amino)carbonyl]-2H tetrazolium hydroxide (XTT) and minimum effective concentration assays in the determination of antifungal susceptibility of Aspergillus fumigatus to the lipopeptide class compounds. AB - The susceptibility of Aspergillus fumigatus to mulundocandin, an echinocandin like compound, and other antifungal agents was assessed by the National Committee for Clinical Laboratory Standards (NCCLS) M38-P method, a 2,3-bis(2-methoxy-4 nitro-5-sulfophenyl)-5-[(phenyl-amino)carbonyl]-2H-tetrazolium hydroxide (XTT) based colorimetric assay, and determination of morphologic alterations by microscopy. In contrast to the NCCLS M38-P method, which does not predict the activity in vivo, the XTT-based assay showed that A. fumigatus is susceptible to mulundocandin. Thus, the XTT-based assay might be useful for determination of the susceptibilities of molds to echinocandins. Further evaluation is warranted. PMID- 11427612 TI - Endophthalmitis caused by Listeria monocytogenes. AB - Listeria monocytogenes was isolated from the aqueous chamber of an immunodepressed patient with acute hypertensive uveitis, who developed a dark hypopyon and pigment dispersion. No extraocular septic focus was found. Treatment was successful with intravitreal vancomycin, anterior chamber irrigation with vancomycin, orally administered ciprofloxacin, and topical fortified vancomycin. PMID- 11427614 TI - Vascular cytochrome P450 in the regulation of renal function and vascular tone: EDHF, superoxide anions and blood pressure. PMID- 11427615 TI - 'Flash pulmonary oedema'--a diagnosis for both the cardiologist and the nephrologist? PMID- 11427616 TI - New aspects in the management of ANCA-positive vasculitis. PMID- 11427617 TI - Compliance with drug therapy-new answers to an old question. PMID- 11427618 TI - Daily dialysis: is this a reasonable option for the new millennium? PMID- 11427619 TI - Renal problems after lung transplantation of cystic fibrosis patients. PMID- 11427620 TI - The epithelial calcium channel, ECaC1: molecular details of a novel player in renal calcium handling. PMID- 11427621 TI - An oral ET(A)-selective endothelin receptor antagonist for contrast nephropathy? PMID- 11427622 TI - The epidemiology of end-stage renal disease in the Baltic countries: an evolving picture. AB - INTRODUCTION: The epidemiology of end-stage renal disease (ESRD) and renal replacement therapy (RRT) is under continuous evolution all over the world. Of particular interest is the development of RRT in the countries of the former Soviet bloc which underwent great political and socio-economical changes in the last decade. We report here the epidemiological analysis of ESRD and RRT in the three Baltic countries: Lithuania, Estonia, Latvia. Subjects and methods. This epidemiological report is based on data from centre questionnaires which were collected from 1996 onwards, with a response rate of 98-99%. RESULTS: The prevalence/incidence of RRT patients in 1999 were 213/99.5 p.m.p. in Lithuania, 186/45.5 p.m.p. in Estonia and 172/55.8 p.m.p. in Latvia. Haemodialysis (HD) was the most common RRT modality in Lithuania (60% of prevalent patients), but not in Estonia (29%), while in Latvia it was nearly as common as renal transplantation (45 and 46%, respectively). Home HD was not performed. The proportion treated by peritoneal dialysis (PD) was very low in Lithuania (4% of RRT patients), while the percentage was higher in Latvia (9%) and Estonia (20.4%). The percentage of patients on RRT treated by renal transplantation was high throughout, representing the main modality of treatment in Estonia (50.5% of RRT prevalent patients, 94 p.m.p.) and in Latvia (46%, 79 p.m.p.) and being high in Lithuania (36%, 77 p.m.p.). The main renal diseases leading to ESRD were glomerulonephritis, pyelonephritis and diabetes. CONCLUSION: The epidemiology of RRT in the Baltic countries is undergoing rapid changes. Transplantation has reached an impressive level. A high percentage of RRT patients live with a functioning graft. PMID- 11427623 TI - Renoprotective and anti-hypertensive effects of combined valsartan and perindopril in progressive diabetic nephropathy in the transgenic (mRen-2)27 rat. AB - BACKGROUND: We have previously reported that severe glomerulosclerosis progressively develops in the streptozotocin (STZ) diabetic transgenic (mRen-2)27 rat. In this diabetic model, monotherapy with either angiotensin converting enzyme inhibition (ACEI) or angiotensin type 1 (AT(1)) receptor blockade is largely renoprotective. The objective of the present study was to determine if a combination therapy at lower doses than monotherapy would confer greater renoprotection. METHODS: At 6 weeks of age, non-diabetic control and STZ diabetic female heterozygous Ren-2 rats were randomized to receive vehicle, the AT(1) receptor blocker valsartan (V, 20 mg/kg/day), the ACEI perindopril (P, 6 mg/kg/day), or a combination of low-dose V+P (V, 3 mg/kg/day plus P, 0.5 mg/kg/day) for 12 weeks. RESULTS: Systolic blood pressure was lowered with all treatments, but the greatest reductions were observed with V monotherapy and combination V+P therapy. All treatments reduced albuminuria, the decline in glomerular filtration rate, and cortical collagen staining, to the same extent. The glomerulosclerotic index was increased with diabetes and reduced with V and P monotherapy. However, the low-dose combination therapy was more effective than single therapy and reduced severe glomerulosclerosis to levels observed in non diabetic controls. CONCLUSIONS: Monotherapy with either V or P reduced blood pressure and retarded the decline in renal function and glomerulosclerosis in the diabetic Ren-2 rat. Combination therapy has the additional benefit of requiring only low doses of AT(1) receptor blockade and ACEI to achieve superior renoprotective effects in this diabetic nephropathy model. PMID- 11427624 TI - Reduction of glomerular hyperfiltration by dopamine D(2)-like receptor blockade in experimental diabetes mellitus. AB - BACKGROUND: Dopamine D(2)-like receptors are involved in the physiological response of renal haemodynamics to amino-acid infusion. The present study was performed to investigate whether domperidone, a D(2)-like receptor antagonist, modulates the pathological hyperfiltration in experimental diabetes mellitus. METHODS: Renal function was studied in anaesthetized rats 2 weeks after induction of moderate diabetes mellitus by streptozotocin, and in non-diabetic controls. Rats in both groups continuously received domperidone or vehicle via drinking water. Following infusion of Ringer's saline for measurement of baseline values, an i.v. amino-acid load was applied to investigate the renal functional reserve. RESULTS: In vehicle-treated diabetic rats baseline glomerular filtration rate and renal plasma flow were significantly higher compared with controls (1.10+/- 0.04 vs. 0.83+/-0.02 (P<0.004) and 4.83+/-0.26 vs 3.32+/-0.24 ml/min/100 g body weight (bw) (P<0.001) respectively). Domperidone completely normalized glomerular filtration rate and renal plasma flow in diabetic rats to values observed in vehicle-treated controls (0.81+/-0.07 (P=0.740) and 3.35+/- 0.30 ml/min/100 g bw (P=0.889) respectively). In the clearance experiments, arterial blood pressure, urinary flow rate and sodium excretion did not significantly differ when comparing the four groups. However, in conscious rats, urinary flow rate and sodium excretion were significantly higher in diabetic rats compared with non diabetic controls. In both non-diabetic groups, amino-acid infusion induced a significant glomerular hyperfiltration that was completely absent in diabetic rats, and restored by domperidone treatment. In conscious vehicle-treated diabetic rats urinary albumin excretion was enhanced (449.0+/-47.7 vs. 185.7+/- 18.1 microg/24 h in non-diabetic rats (P<0.001)) and significantly lowered in diabetic rats by domperidone treatment (109.8+/-15.4 microg/24 h (P<0.001)). CONCLUSION: The data suggest that dopaminergic mechanisms are involved in the changes in renal haemodynamics during early experimental diabetes mellitus in rats. PMID- 11427625 TI - High proteinuria selectivity index based upon IgM is a strong predictor of poor renal survival in glomerular diseases. AB - BACKGROUND: The transport of large proteins across the glomerular capillary wall (GCW) may increase several fold in glomerular diseases. The occurrence of IgM in urine is a consequence of the presence of large defects or shunts in the GCW, whereas albuminuria is probably a result of an altered charge- and size selectivity of the GCW. In order to examine whether patho-morphological differences influence the renal outcome in proteinuric glomerulopathies, we examined urinary excretion of IgM and albumin as prognostic markers of glomerular disease. METHODS: An observational study over a median of 41 (+/-3) months was conducted in 84 patients with biopsy-verified glomerular disease. The patients were subdivided into groups with low (< or =0.002) and high (>0.002) proteinuria selectivity index based upon IgM (IgM-SI), and into groups with low (< or =200 mg/mmol) and high (>200 mg/mmol) albumin creatinine index (ACI). RESULTS: In the high IgM-SI group, the median creatinine clearance (Ccr) decreased by 26%, and 62% of the patients decreased in Ccr by >5 ml/ min/year during the follow-up time. In comparison, the median Ccr decreased by 8% in the low IgM-SI group (P<0.001) and only 18% of the patients in this group deteriorated by >5 ml/min/year in the Ccr. Eleven (21%) of the 51 patients in the high IgM-SI group developed end-stage renal failure compared with none of the 33 patients in the low IgM-SI group. All the patients that progressed to uraemia had decreased Ccr (<60 ml/min) at entry into the study. However, among all these patients, only those with high IgM-SI, and none with low IgM-SI, developed end stage renal failure. The fall in Ccr did not differ significantly between the patients in high (12%) and low (16%) ACI groups. CONCLUSION: The results of this study indicate that an increased IgM-SI value is a stronger predictor of clinical outcome in proteinuric glomerulopathies than baseline albuminuria. This finding may reflect different patho-histological mechanisms influencing renal survival in glomerular diseases. PMID- 11427626 TI - The incidence of biopsy-proven glomerulonephritis in Australia. AB - BACKGROUND: There is limited population-based epidemiological data on renal disease. An insight into the spectrum of clinically significant glomerulonephritis can be obtained from renal biopsy diagnoses. This is a descriptive report of biopsy-proven glomerulonephritis within a defined population. METHODS: A retrospective review of the pathology reports of all native renal biopsies performed in the Australian state of Victoria in 1995 and 1997 was undertaken. Trends in the average annual age- and sex-specific incidence rates for biopsy-proven glomerulonephritis were calculated. Comparisons were made with the incidence of end-stage renal disease due to glomerulonephritis confirmed on renal biopsy. RESULTS: The most common glomerulonephritides in adults are IgA disease, focal glomerulosclerosis, lupus nephritis and vasculitis, and in children are lupus nephritis, focal glomerulosclerosis, IgA disease and minimal change disease. A male predominance is seen for all glomerulonephritides, except lupus nephritis, in both adults and children. An increase in incidence of disease with age, particularly in males, is seen for vasculitis and focal glomerulosclerosis. The most common glomerulonephritides on renal biopsy are reflected in the most common causes of end-stage renal disease due to glomerulonephritis. CONCLUSIONS: This review has provided population-based descriptive epidemiological data on clinically significant glomerulonephritis. This data provides important clues for further studies relating to the identification of risk factors for the various types of glomerulonephritis. PMID- 11427627 TI - Polymorphism of the cytokine genes in hospitalized patients with Puumala hantavirus infection. AB - BACKGROUND: Nephropathia epidemica (NE) is a mild type of haemorrhagic fever with renal syndrome caused by Puumala (PUU) hantavirus. The clinical course of NE varies from asymptomatic to fatal. The aim of this study was to establish whether polymorphisms in the cytokine genes are associated with susceptibility to and outcome of NE. METHODS: The genotypes of the genes of tumour necrosis factor alpha (TNFalpha), interleukin-1alpha (IL-1alpha), IL-1beta and IL-1 receptor antagonist (IL-1RA) were analysed by polymerase chain reaction in 87 subjects, all hospital-treated for serologically confirmed acute NE. The control group comprised 400 healthy blood donors. Nineteen out of these 400 (5%) controls were PUU virus-seropositive. RESULTS: IL-1RA allele 2 and IL-1beta (base exchange polymorphism at position -511) allele 2 were strongly associated with each other in both groups. NE patients were more often IL-1RA-2 negative/IL-1beta-2 negative than PUU-seronegative blood donors (38 vs 27%, odds ratio 1.65, 95% confidence interval 1.0-2.7). However, there were no differences in the clinical severity of NE between the IL-1RA-2 negative/IL-1beta-2 negative and the other patients. The other allele frequencies studied evinced no statistically significant differences between the groups. Thirty-three out of 87 (38%) patients and 121 out of 381 (32%) seronegative controls were carriers of the high-producer genotype TNF2 allele. Several parameters showed the clinical course of NE to be more severe in TNF2 carriers than in non-carriers. CONCLUSIONS: These data suggest that non carriage of the IL-1RA allele 2 and IL-1beta (-511) allele 2 may contribute to susceptibility to NE. Furthermore, TNFalpha polymorphism seems to be associated with the outcome of NE. PMID- 11427628 TI - T cell receptor BV gene usage in interstitial cellular infiltrates in active Heymann nephritis. AB - BACKGROUND: Infiltration of the kidney by mononuclear cells is a prominent feature of active Heymann nephritis (HN). These cells could be present as a part of generalized inflammatory response, or could be proliferating in response to specific antigens. To examine these questions, we have analysed the T cell receptor (TCR) BV repertoire of T cells infiltrating the renal interstitium at regular time intervals throughout the course of the disease. METHODS: HN was induced in Lewis rats by immunization with renal tubular antigen (Fx1A) in complete Freund's adjuvant (CFA). Kidneys were collected 8 and 12 weeks after immunization. Renal tissue was homogenized and RNA extracted. RT-PCR and sequencing were used to characterize expression of TCR BV genes. RESULTS: Preferential expression of TCR BV2 and BV16 gene families was seen at 8 weeks. By 12 weeks the diversity of the TCR BV gene repertoire had increased and was highly heterogeneous. Sequence analysis of BV2, and BV16 RT-PCR products from 8 week HN kidneys revealed conserved usage of CDR3 regions, and an over-representation of arginine residues in the CDR3 regions at a frequency of between 60 and 100% of clones sequenced in most of BV2 and BV16 subfamilies. CONCLUSION: The preferential usage of CDR3 region sequences in TCR BV2 and BV 16 families indicates clonal expansion of individual T cells in HN kidneys at 8 weeks. The conserved usage of arginine residues in the CDR3 regions may indicate recognition of select antigenic epitopes. By 12 weeks, the diverse TCR BV repertoire in the kidney may be due to epitope spreading or may represent a non-specific inflammatory response in the late phase of the disease. PMID- 11427629 TI - Urinary albumin excretion rate and glomerular filtration rate in the prediction of diabetic nephropathy; a long-term follow-up study of childhood onset type-1 diabetic patients. AB - BACKGROUND: Predictors of diabetic nephropathy are only partly known. The role of glomerular hyperfiltration is much discussed. We have studied the cumulative incidence of micro and macroalbuminuria and the predictive value of glomerular filtration rate (GFR) and screening value of albumin excretion rate (AER) in type 1 diabetes. METHODS: A cohort of diabetic children was followed up at a mean duration of 29+/-3 years. All 75 children treated in one hospital with diabetes duration > or =8 years were prospectively followed for 8 years examining GFR, AER, blood pressure and HbA1c. After another 8-10 years, 60 of them were traced for endpoint follow-up. RESULTS: Seven patients (12%) developed macroalbuminuria, i.e. persistent overnight AER>200 mg/min, 12 (20%) developed persistent microalbuminuria (AER 15-200 mg/min) and 17 (28%) transient microalbuminuria (>15 mg/min on two consecutive occasions, normalized at endpoint). One baseline screening value of 24-h AER>15 mg/min predicted 93% of patients with persistent micro or macroalbuminuria. The negative predictive value was 78%. Six of seven macroalbuminuric and 10 of 12 microalbuminuric patients had a baseline GFR above the normal limit of the method (> or =125 ml/min/1.73 m(2)). When adjusted for diabetes duration, increased GFR predicted macro or microalbuminuria (odds ratios=5.44, P=0.04). The positive predictive value for having an increased baseline GFR was 53%. The negative predictive value was 77%. Stratification for HbA1c did not change the effect of an increased GFR. CONCLUSIONS: At a mean diabetes duration of 29 years the cumulative incidence of macroalbuminuria was 12%; however, another 20% had persistent microalbuminuria. A screening value of 24-h AER >15 mg/min was a strong predictor, whereas increased GFR was a weaker but significant predictor for micro and macroalbuminuria. PMID- 11427630 TI - Self-assessed physical and mental function of haemodialysis patients. AB - BACKGROUND: Physical (PCS) and mental (MCS) component summary scales of the Short Form 36 (SF-36) health survey are validated measures of quality of life (QOL) and functional status. We sought to evaluate the PCS and MCS in haemodialyis patients as compared to the general population and other chronic diseases. METHODS: A cohort of 134 haemodialysis patients (mean age 60.9+/-14.3 years, males 63.4%, Caucasians 66.4%) was followed from January 1996 to December 1998 (mean follow up 14.5+/-5.7 months). SF-36 questionnaires were administered every 3 months and PCS and MCS were calculated. Results were compared to the general population and other chronic diseases. Correlators of PCS and MCS, change in QOL over time, and the correlators of this change were determined. RESULTS: Mean PCS was 36.9+/-8.8 and mean MCS was 47+/-10.7. Compared to the general US population, these represent a decline of 8.7+/-0.8 for PCS (P<0.0001) and 2.7+/-0.8 for MCS (P<0.001). PCS and MCS in end-stage renal disease (ESRD) were lower than in most other chronic diseases studied. Univariate correlators of PCS in haemodialysis patients included age, male sex, haematocrit, serum albumin, and severity of comorbid cardiac and pulmonary illnesses. Multivariate analysis demonstrated independent correlators of PCS to be male sex, serum albumin and severity of comorbid cardiac and pulmonary diseases. Univariate as well as multivariate correlators of MCS included: serum albumin, KT/V(urea), and status living alone. A trend analysis revealed that both PCS and MCS tended to decline in the initial months of dialysis but stabilized over time. Status living alone was a significant predictor of improvement in MCS by univariate as well as multivariate analysis. CONCLUSIONS: Self assessed physical and mental health of haemodialysis patients is markedly diminished compared to the general population and other chronic diseases. PMID- 11427631 TI - Does ethnicity influence perceived quality of life of patients on dialysis and following renal transplant? AB - BACKGROUND: Quality of life (QoL) as perceived by patients with end-stage renal disease (ESRD) is an important measure of patient outcome. There is a high incidence of ESRD in the Indo-Asian population in the UK and a lower rate of transplantation compared with white Europeans. The aim of this study was to determine whether perceived quality of life was influenced by treatment modality and ethnicity. METHODS: Sixty Indo-Asians treated with either peritoneal dialysis (n=20), hospital haemodialysis (n=20) or with a renal transplant (n=20) for >3 months were compared with 60 age-matched white Europeans closely matched for gender, diabetes and duration of renal replacement therapy. QoL was measured using the Kidney Disease and Quality of Life questionnaire (KDQOL-SF). The KDQOL SF measures four QoL dimensions: physical health (PH), mental health (MH), kidney disease-targeted issues (KDI) and patient satisfaction (PS). Adequacy of treatment was measured by biochemistry, 24 h urine collection and dialysis kinetics. The number of comorbid conditions was scored. Social deprivation was calculated from the patient's postal address using Townsend scoring. RESULTS: QoL was significantly lower in Indo-Asians than white Europeans for PH, MH and KDI. This was not related to treatment adequacy, which was similar in both for each modality. Indo-Asians had a worse index of social deprivation than white Europeans (P=0.008). PH and KDI were related to social deprivation (P=0.007 and P=0.005, respectively). QoL (except PS) was inversely correlated with comorbidity. Dialysis patients had higher comorbidity than transplant patients (P<0.02). Comparing only those dialysis patients considered fit for transplantation (n=51) with transplant patients, comorbidity was similar, but differences in QoL persisted. CONCLUSION: This study demonstrates a lower perceived QoL in Asians compared with white Europeans with ESRD. Analysis of QoL indicates that Asian patients in particular perceive kidney disease as a social burden, even if successfully transplanted. PMID- 11427632 TI - Strong depletion of CD14(+)CD16(+) monocytes during haemodialysis treatment. AB - BACKGROUND: The immune defect in haemodialysis (HD) patients is associated with a monocytic dysfunction, including an increased production of proinflammatory cytokines. Monocytes fall into subpopulations comprising CD14(++)CD16(-) and CD14(+)CD16(+) cells. Circulating numbers of the latter can rapidly increase during infectious episodes and inflammation. METHODS: We determined the amount of CD14(+)CD16(+) monocytes in HD patients and characterized their fate during HD treatment. In 34 HD patients and 17 healthy controls, the distinct cell populations were determined by differential blood counts and flow cytometry. Cells from 14 HD patients were analysed at the start, 10, 30 and 120 min thereafter, and at the end of HD treatment. RESULTS: Before HD, patients show a monocytosis with a strongly increased CD14(+)CD16(+) subpopulation. Early during HD treatment, circulating leukocyte numbers decrease, with monocytes being most profoundly influenced. Interestingly, among them, sequestration is most pronounced in the CD14(+) CD16(+) subpopulation. After 30 min, approximately 83+/ 9% of CD14(+)CD16(+) cells are removed from circulation. This sequestration does not differ between patients treated with polyamide or haemophan membranes. The sequestration is a short-lived temporary effect and cell numbers are replenished within 120 min of treatment for the entire monocyte population. Beyond that time point, cellular activation by the dialyser membrane becomes visible. Reappearence kinetics of CD14(+)CD16(+) monocytes is slower; however, initial numbers are reached by the end of treatment. CONCLUSION: Haemodiaysis leads to temporary removal of monocytes from the bloodstream followed by the reappearance of activated cells. This might contribute to the state of chronic microinflammation, which is reflected by high levels of CD14(+)CD16(+) monocytes. PMID- 11427633 TI - Which parameters affect cytosolic free calcium in polymorphonuclear leukocytes of haemodialysis patients? AB - BACKGROUND: Cytosolic free calcium ([Ca(2+)](i)) is an important second messenger during stimulation in a wide variety of cells, including polymorphonuclear leukocytes (PMNs). Its mobilization in PMNs is altered in various diseases such as atherosclerosis and ageing. In chronic haemodialysis (HD) patients, both atherosclerosis and accelerated ageing are well known. Therefore [Ca(2+)](i) in resting PMNs of HD patients was determined along with certain parameters which might affect it, such as recombinant human erythropoietin (rHuEpo) treatment, calcium-phosphate balance, and biocompatibility of dialysis membranes. METHODS: PMNs were separated by density centrifugation and [Ca(2+)](i) was determined by spectrofluorimetry using Quin 2/AM fluorescent dye. Laboratory parameters were determined by standard methods in clinical chemistry. RESULTS: It was found that [Ca(2+)](i) in resting PMNs of HD patients not undergoing rHuEpo therapy was higher than that of controls. After 12-weeks of rHuEpo therapy, [Ca(2+)](i) decreased to near normal level. The role of erythropoiesis in normalization of [Ca(2+)](i) in resting PMNs was supported by PMN [Ca(2+)](i) which was elevated in patients who had low haemoglobin (<100 g/l) or haematocrit (<0.30) values. In some patients, including those receiving rHuEpo treatment, [Ca(2+)](i) remained high, suggesting a role for other parameters in increasing [Ca(2+)](i). One possible parameter might be the disturbed calcium-phosphate metabolism of chronic renal failure, because we found a strong correlation between [Ca(2+)](i) and plasma iPTH levels in HD patients (r=0.743, P<0.001). [Ca(2+)](i) was also elevated in PMNs of those patients who had either low plasma calcium or high plasma phosphate levels. PMN [Ca(2+)](i) of HD patients correlated positively with the duration of HD (r=0.671, P<0.001). However, there was no correlation between [Ca(2+)](i) and patient age. The dialysis procedure itself also transiently increased PMN [Ca(2+)](i) HD patients, independently of the type of dialysis membrane. CONCLUSION: PMN [Ca(2+)](i) is modulated by various parameters in HD patients, including the degree of anaemia, disturbances of calcium metabolism, and duration of dialysis treatment. The elevated [Ca(2+)](i) of resting PMNs might contribute to altered functions in these cells. PMID- 11427634 TI - The role of iron status markers in predicting response to intravenous iron in haemodialysis patients on maintenance erythropoietin. AB - BACKGROUND: Iron deficiency (ID) is the main cause of hyporesponsiveness to erythropoietin in haemodialysis patients and its detection is of value since it is easily corrected by intravenous iron. Markers of iron supply to the erythron, including erythrocyte zinc protoporphyrin (Er-ZPP), percentage of hypochromic erythrocytes (Hypo), reticulocyte haemoglobin content (CHr) and soluble transferrin receptor (sTfR), may be more accurate predictors of ID than ferritin (Fer) and transferrin saturation (TSat), but relative diagnostic power and best threshold values are not yet established. METHODS: In 125 haemodialysis patients on maintenance erythropoietin, the diagnostic power of the above parameters was evaluated by ROC curve, multivariate regression, and stepwise discriminant analyses. Diagnosis of ID was based on haemoglobin response to intravenous iron (992 mg as sodium ferric gluconate complex over an 8-week period). RESULTS: Fifty one patients were considered iron deficient (haemoglobin increase by 1.9+/-0.5 g/dl) and 74 as iron replete (haemoglobin increase by 0.4+/-0.3 g/dl). ROC curve analysis showed that all tests had discriminative ability with the following hierarchy: Hypo (area under curve W=0.930, efficiency 89.6% at cut-off >6%), CHr (W=0.798, efficiency 78.4% at cut-off < or =29 pg), sTfR (W=0.783, efficiency 72.4% at cut-off >1.5 mg/l), Er-ZPP (W=0.773, efficiency 73.0% at cut-off >52 micromol/mol haem), TSat (W=0.758, efficiency 70.4% at cut-off <19%) and ferritin (W=0.633, efficiency 64.0% at cut-off <50 ng/ml). Stepwise discriminant analysis identified Hypo as the only variable with independent diagnostic value, able to classify 87.2% of patients correctly. Additional tests did not substantially improve diagnostic efficiency of Hypo >6% alone. CONCLUSIONS: In haemodialysis patients on maintenance erythropoietin, Hypo >6% is the best currently available marker to identify those who will improve their response after intravenous iron. Cost-effectiveness suggests that this parameter should be a first-line tool to monitor iron requirements in clinical practice. PMID- 11427635 TI - A novel technique to demonstrate disturbed appetite profiles in haemodialysis patients. AB - BACKGROUND: Malnutrition is common among dialysis patients and is associated with an adverse outcome. One cause of this is a persistent reduction in nutrient intake, suggesting an abnormality of appetite regulation. METHODS: We used a novel technique to describe the appetite profile in 46 haemodialysis (HD) patients and 40 healthy controls. The Electronic Appetite Rating System (EARS) employs a palmtop computer to collect hourly ratings of motivation to eat and mood. We collected data on hunger, desire to eat, fullness, and tiredness. HD subjects were monitored on the dialysis day and the interdialytic day. Controls were monitored for 1 or 2 days. RESULTS: Temporal profiles of motivation to eat for the controls were similar on both days. Temporal profiles of motivation to eat for the HD group were lower on the dialysis day. Mean HD scores were not significantly different from controls. Dietary records indicated that dialysis patients consumed less food than controls. CONCLUSIONS: Our data indicate that the EARS can be used to monitor subjective appetite states continuously in a group of HD patients. A HD session reduces hunger and desire to eat. Patients feel more tired after dialysis. This does not correlate with their hunger score, but does correlate with their fullness rating. Nutrient intake is reduced, suggesting a resetting of appetite control for the HD group. The EARS may be useful for intervention studies. PMID- 11427636 TI - Quantitative flow cytometry shows activation of the TNF-alpha system but not of the IL-2 system at the single cell level in renal replacement therapy. AB - BACKGROUND: Immunological dysfunction in patients on haemodialysis may be related to imbalanced cytokine systems, such as tumour necrosis factor (TNF)-alpha and interleukin (IL)-2. Despite activation of these systems, haemodialysis patients show high susceptibility for infections and malignancies, and have a poor immunological reaction to T-cell-dependent antigens, like hepatitis B vaccination. In this study we have determined the activation status of the two different cytokine systems, at the single cell level, using quantitative flow cytometry. METHODS: Using fluorescein isothiocyanate- or phycoerythrin-conjugated antibodies directed against TNF-R2 (CD120b), IL-2Ralpha (CD25) and IL-2Rbeta (CD122), we measured the expression of these receptors at the single cell level in order to determine the level of activation of monocytes and T-lymphocytes. RESULTS: Significantly higher expression of the TNF-alpha receptor, TNF-R2, was present on both monocytes and T-lymphocytes in patients on renal replacement therapy (RRT) compared with pre-dialysis chronic renal failure (CRF) patients and controls, indicating activation of the TNF-alpha system. In contrast, IL-2R expression was comparable in all groups studied, which may reflect a non activated state of the IL-2 system. CONCLUSIONS: The present study illustrates an activated state of the TNF-alpha system in patients on RRT, at the single cell level, while the IL-2 system seems to be unaffected. These findings support the hypothesis that the interaction between the TNF-alpha and IL-2 cytokine systems is disturbed. PMID- 11427637 TI - Methylene blue, a nitric oxide inhibitor, prevents haemodialysis hypotension. AB - BACKGROUND: Plasma nitric oxide (NO) levels have been found to be high in haemodialysis (HD) patients, especially in those prone to hypotension in dialysis. The aim of the study was to prevent dialysis hypotension episodes by i.v. administration of methylene blue (MB), an inhibitor of NO activity and/or production. METHODS: MB was given i.v. in 18 stable HD patients with hypotensive episodes during almost every dialysis, in 18 HD patients without hypotension during dialyses, and in five healthy controls. MB was given as a bolus of 1 mg/kg bodyweight followed by a constant infusion of 0.1 mg/kg bodyweight lasting 210 min until the end of the dialysis session and only as a bolus on a non-dialysis day. Systolic and diastolic blood pressures (BP) were measured at 10-min intervals during HD sessions with or without MB and on a non-dialysis day with MB. RESULTS: In hypotension-prone patients, MB completely prevented the hypotension during dialysis and increased both systolic and diastolic BP on non dialysis days. In normotensive patients, MB increased BP during the first hour of dialysis and for 90 min on the non-dialysis day. The BP in the healthy controls remained unchanged. Plasma and platelet NO(2)+NO(3) (stable metabolites of NO) levels were determined. The NO(2)+NO(3) generation rate in the first post dialysis day was calculated. The plasma and platelet NO(2)+NO(3) were higher in the hypotensive group than in the normotensive dialysis group. The generation rate of nitrates was higher (P<0.01) in the hypotensive group (1.21+/-0.13 micromol/min and 0.74+/-0.16 after MB) than in the normotensive patients (0.61+/ 0.11 micromol/ min and 0.27+/-0.14 after MB). No side-effects were recorded. CONCLUSIONS: MB is an efficient therapy in the prevention of dialysis hypotension. PMID- 11427638 TI - Endothelial dysfunction marker von Willebrand factor antigen in haemodialysis patients: associations with pre-dialysis blood pressure and the acute phase response. AB - BACKGROUND: Increased plasma soluble von Willebrand factor antigen (vWF : Ag) level, a marker of vascular endothelial cell dysfunction, is a strong predictor of atherosclerotic cardiovascular disease (CVD) in the general population. We studied cross-sectional associations between vWF : Ag level, prevalence of CVD, and related factors including pre-dialysis arterial blood pressure (BP) and some markers of inflammation in maintenance haemodialysis (HD) patients. Methods and results. Plasma vWF : Ag level measured by an enzyme-linked immunosorbent assay (ELISA) was higher in 110 HD patients than in 20 controls. On bivariate regression analysis, vWF : Ag level was directly associated with the presence of CVD, age, fibrinogen and the use of enoxaparin (vs unfractionated heparin) during HD procedures, and inversely with albumin and pre-dialysis BP. The patients with prevalent CVD were older, had higher vWF : Ag, white blood cell and platelet counts, fibrinogen and triglycerides, lower albumin levels, and were less frequently on combination antihypertensive therapy. Multivariable analyses identified low pre-dialysis BP, hypoalbuminaemia and hyperfibrinogenaemia (in descending order of significance) as independent predictors of high vWF : Ag level. There were no associations between vWF : Ag levels and gender, ABO blood type, smoking, body mass index, renal failure cause, duration of HD therapy, K(t)/V, normalized protein catabolic rate, dialysate buffers, dialysers, viral hepatitis, erythropoietin treatment, specific antihypertensive drugs, haemoglobin, white blood cell and platelet counts, liver enzymes, phosphorous, total cholesterol, and triglycerides. CONCLUSION: Elevated plasma levels of endothelial dysfunction marker vWF : Ag in maintenance HD patients are associated with established cardiovascular mortality risk factors such as low pre-dialysis blood pressure and the activated acute phase response. PMID- 11427639 TI - A randomized, double-blind, placebo-controlled trial of supplementary vitamins E, C and their combination for treatment of haemodialysis cramps. AB - BACKGROUND: Muscle cramps that improve after carnitine or vitamin E therapies are common in haemodialysis (HD) patients. Because vitamin C participates in carnitine biosynthesis, and its levels are reduced in uraemia, subclinical vitamin C depletion may contribute to HD cramps. Our aim was to determine the effects of vitamins C, E and their combination on the frequency and intensity of HD cramps. METHODS: In this placebo-controlled, double-blind study, 60 HD patients were randomized into four therapeutic groups. Each group (n=15) received six identical capsules daily for 8 weeks, containing one of the following: vitamin E (400 mg), vitamin C (250 mg), their combination, or placebo. RESULTS: The frequency and intensity of HD cramps decreased significantly in all three vitamin groups compared with the placebo group at the end of the trial, and compared with the pre-treatment values. At the end of the trial, vitamins E, C, their combination, and placebo produced cramp reductions of 54, 61, 97 and 7%, respectively. The percentage cramp reduction had no significant correlation with age, sex, aetiology of end-stage renal disease, serum electrolytes or HD duration, but showed a positive correlation (r=0.33, P=0.01) with the type of therapy. No vitamin-related adverse effects were encountered during the trial. CONCLUSION: Short-term treatment with the combination of vitamins E and C is safe and effective in reducing HD cramps; however, its safety for prolonged therapy has yet to be evaluated in HD patients. PMID- 11427640 TI - Factors associated with increased serum levels of cardiac troponins T and I in chronic haemodialysis patients: Chronic Haemodialysis And New Cardiac Markers Evaluation (CHANCE) study. AB - BACKGROUND: Serum concentrations of the cardiac troponins (cTn) T and I, specific markers of myocardial injury, are frequently elevated in haemodialysis patients. The clinical relevance of this is unclear. The aim of this study was to investigate factors associated with increased serum levels of cTn in haemodialysis patients. METHODS: We included in this cross-sectional study 258 chronic haemodialysis patients (150 men, age 60+/-15 years) without acute coronary symptoms. Clinical data, echocardiographic hypertrophy, biochemical status, and haemodialysis regimen were evaluated for each patient. Pre-dialysis serum cTnT (Elecsys, Roche), cTnI (Stratus and RXL, Dade-Berhing), and CK-MB (Stratus, Dade-Berhing) concentrations were determined. Logistic regression was the principal method of analysis. RESULTS: Pre-dialysis levels of cTnT >0.1 ng/ml (n=48, 18.6% of patients) were associated with age (P<0.001), diabetes (P<0.005), history of ischaemic heart disease (P<0.05), and left ventricular hypertrophy (P<0.05). In multivariate analysis, age odds ratio ((OR) 1.04), diabetes (OR 4.9), and indexed left ventricular mass (OR 1.01) were found to be independently associated with cTnT concentration above the threshold. Only six patients had cTnI-Stratus levels >0.6 ng/ml. cTnI-RXL levels >0.3 ng/ml (n=13, 5.0%) were associated with age (P=0.05) and hypercholesterolaemia (P<0.05). Only age (OR 1.06) remained associated in multivariate analysis. CONCLUSION: Elevated baseline serum levels of cardiac troponins were associated with cardiovascular risk factors, history of ischaemic heart disease and left ventricular hypertrophy in asymptomatic chronic haemodialysis patients. PMID- 11427641 TI - Long-term CAPD patients are volume expanded and display more severe left ventricular hypertrophy than haemodialysis patients. AB - BACKGROUND: Whether hypertension and left ventricular hypertrophy (LVH) are more prevalent in CAPD than in haemodialysis (HD) patients is still under discussion. METHODS: To examine this problem we compared a group of 51 CAPD patients, with a group of 201 HD patients. The evaluation included the measurement of atrial natriuretic peptide (atrial natriuretic factor (ANF)), taken as indicator of volume status, and echocardiographic measurements. RESULTS: CAPD patients were older, had been treated for a shorter time, and had lower serum albumin and phosphate than HD patients. Plasma ANF was higher (P<0.01) in CAPD (median 33.8 pmol/l (interquartile range 18.2-63.0)) than in HD patients (22.7 pmol/l (14.9 38.7)). Similarly, the left atrial volume was substantially higher (P<0.0001) in CAPD patients (49+/-22 ml) than in HD patients (37+/-17 ml), while the left ventricular end-diastolic diameter was similar in the two groups (CAPD 51+/-7 mm; HD 50+/-7 mm). Furthermore, left ventricular hypertrophy was more severe (P<0.0001) in CAPD (157+/-37 g/m(2)) than in HD patients (133+/-39 g/m(2)). The proportion of CAPD patients requiring antihypertensive drugs was markedly higher than that of HD patients (65 vs 38% P<0.001). Multivariate modelling showed that volume expansion and pressure load as well as serum albumin were independent predictors of left ventricular mass. CONCLUSIONS: Left ventricular hypertrophy is more severe in long-term CAPD patients than in HD patients. This finding is associated with evidence of more pronounced volume expansion, hypertension, and hypoalbuminaemia. Volume and pressure load along with factors associated with hypoalbuminaemia may aggravate LVH in uraemic patients on CAPD. PMID- 11427642 TI - Better microvascular function on long-term treatment with lisinopril than with nifedipine in renal transplant recipients. AB - BACKGROUND: The prevalence of hypertension in renal transplant recipients is high but the pathophysiology is poorly defined. Impaired endothelial function may be a factor of major importance. The present study addresses the effects of long-term treatment with either lisinopril or slow-release nifedipine on microvascular function and plasma endothelin in renal transplant recipients on cyclosporin A (CsA). METHODS: Seventy-five hypertensive renal transplant recipients were double blind randomized to receive slow-release nifedipine (NIF, n=40) or lisinopril (LIS, n=35). Ten normotensive, age-matched recipients served as controls. All patients received CsA-based immunosuppressive therapy including prednisolone and azathioprine. Microvascular function was assessed in the forearm skin vasculature, using laser Doppler flowmetry in combination with post-occlusive reactive hyperaemia and endothelial-dependent function during local acetylcholine (ACh) stimulation. RESULTS: The analysis of microvascular function (AUC(rh)) showed that nifedipine-treated patients had significantly lower responses compared with lisinopril-treated patients (20+/-17 and 43+/-20 AU x min respectively, P=0.0016). Endothelial function was borderline significantly lower in the NIF group compared with the LIS group (640+/-345 and 817+/-404 AU x min respectively, P=0.056). The responses in the LIS group were comparable with those in non-hypertensive controls (AUC(rh) was 37+/-16 and AUC(ACh) was 994+/-566 AU x min). Plasma endothelin-1 concentrations were significantly higher in the NIF group compared with the LIS group (0.44+/-0.19 vs. 0.34+/-0.10 fmol/ml respectively, P=0.048), and were 0.29+/-0.09 fmol/ml in the control patients. AUC(ACh) was associated with plasma endothelin-1 (P=0.0053), while AUC(rh) was not (P=0.080). CONCLUSIONS: The study indicates that long-term treatment with lisinopril, when compared with nifedipine, yields a more beneficial effect on microvascular function in hypertensive renal transplant recipients on CsA. The beneficial microvascular effect may be mediated in part by an endothelin-1 associated effect on the endothelium. PMID- 11427643 TI - Removal of contrast media by different extracorporeal treatments. AB - BACKGROUND: Although the capability of extracorporeal treatments after administration of contrast media to prevent radiocontrast-induced nephropathy is controversial, haemodialysis is performed in many institutions after radiographic procedures. There are conflicting reports on the efficacy of different dialysers and treatment modalities to remove contrast media. METHODS: We compared the contrast medium-removing ability of different extracorporeal treatments in a randomized trial. Thirty-nine patients on chronic renal-replacement therapy or with chronic renal failure were randomized to receive low-flux haemodialysis (Low HD, n=10), high-flux haemodialysis (High-HD, n=10), online haemodiafiltration (HDF, 10 litre substitution, n=10) and online haemofiltration (HF, 18 litre substitution, n=9) after administration of contrast medium during routine radiological procedures. Plasma concentrations of contrast medium (iopromide or iomeprol) were measured by energy-dispersive X-ray fluorescence analysis. RESULTS: The extraction ratio for contrast media was 0.64+/-0.1 for Low HD (P<0.05 vs. High-HD and vs. HDF), 0.74+/-0.1 for High-HD (P<0.05 vs. HF), 0.81+/ 0.1 for HDF (P<0.05 vs HF), and 0.62+/-0.1 for HF. Mean extracorporeal plasma clearances were 82+/-2 for Low-HD (P<0.05 vs. High-HD and vs HDF), 100+/-2 for High-HD, 115+/-4 for HDF (P<0.05 vs. HF), and 86+/-5 ml/min for HF. CONCLUSIONS: We conclude that HDF and High-HD remove contrast media more effectively than Low HD and HF during the time of each treatment session. However, whether this is also true for the overall elimination of contrast media by these different procedures needs to be addressed in future studies, by a precise assessment of the drug time course after the session. PMID- 11427644 TI - Castleman's disease with nephrotic syndrome, amyloidosis and autoimmune manifestations. PMID- 11427645 TI - Severe non-proliferative lupus nephritis with predominant sub-endothelial IgA deposits. PMID- 11427646 TI - Valproic acid overdose and haemodialysis. PMID- 11427647 TI - Primary bilateral T-cell renal lymphoma presenting with sudden loss of renal function. PMID- 11427648 TI - Unclear radiographic pulmonary changes in a patient who recently underwent renal transplantation. PMID- 11427649 TI - Right-sided chest pain at the onset of haemodialysis. PMID- 11427650 TI - 'Decoy cells' in the urine due to polyomavirus BK infection: easily seen by phase contrast microscopy. PMID- 11427651 TI - Acute renal failure in a young woman with endometriosis. PMID- 11427652 TI - Severe hypokalaemia and paralysis in a Chinese scientist. PMID- 11427653 TI - Acquired renal cysts. PMID- 11427654 TI - Reflections on the XVth ICN meeting: renal and cardiovascular protection in the developing world. PMID- 11427655 TI - Dialysis membranes in convective treatments. PMID- 11427656 TI - An unusual presentation of relapsing Wegener's granulomatosis. PMID- 11427657 TI - Dent's disease: can we slow its progression? PMID- 11427658 TI - Immunoglobulin as a risk factor for contrast media nephrotoxicity. PMID- 11427659 TI - Effect of N-acetylcysteine on serum creatinine level. PMID- 11427660 TI - Coexisting renal artery stenosis and membranous glomerulonephritis. Is there a link? PMID- 11427661 TI - Fibrate-induced increase in blood urea and creatinine. PMID- 11427662 TI - Renal failure and nephrotic syndrome associated with gastrointestinal stromal tumour (GIST)-a rare cause of AA amyloidosis. PMID- 11427663 TI - Renal bone disease in black dialysis patients: are algorithms developed for white dialysis patients valid? PMID- 11427664 TI - Using AN69 ST membrane: a dialysis centre experience. PMID- 11427665 TI - Iliac vein stenosis secondary to femoral catheter placement. PMID- 11427666 TI - Trisodium citrate 30% vs. heparin 5% as catheter lock in the interdialytic period in twin- or double-lumen dialysis catheters for intermittent haemodialysis. PMID- 11427667 TI - Antibiotic management of infected vascular access. PMID- 11427668 TI - Does ultrapure dialysate prevent the development of biofilm in dialysis therapy? PMID- 11427669 TI - Renal failure, anaemia, cytokines and inflammation. PMID- 11427670 TI - Epoietin-alpha-associated total alopecia. PMID- 11427671 TI - Traumatic pleural leak in peritoneal dialysis. PMID- 11427673 TI - Visualization of an FtsZ ring in chloroplasts of Lilium longiflorum leaves. AB - FtsZ is a bacterial division protein which forms a ring at the leading edge of the cell division site. To date, a hypothesis that the plant FtsZ forms the same structure in chloroplast division is proposed, but has not been demonstrated yet. In this study, recombinant LlFtsZ (Lilium longiflorum FtsZ) protein was produced from a previously isolated ftsZ cDNA clone [Mori and Tanaka (2000) Protoplasma 214: 57] and used to raise polyclonal anti-LlFtsZ antibodies in rabbits. In immunoblot analysis with the total protein extracted from L. longiflorum leaves, purified antibodies specifically recognized LlFtsZ whose molecular mass was approximately 43 kDa. This size corresponded to that of the recombinant LlFtsZ protein lacking N-terminal sequence, which suggests that the full-length LlFtsZ translation product has a putative N-terminal signal peptide. Moreover, fluorescent and electron microscopy revealed that the anti-LlFtsZ antibodies recognized ring structures at stromal side of the constriction point of dividing chloroplasts. Here, we show direct evidence that FtsZ ring is involved in chloroplast division. PMID- 11427674 TI - Molecular aspects of self-incompatibility in Brassica species. AB - Many flowering plants possess self-incompatibility (SI) systems to prevent inbreeding. SI in Brassica species is controlled by a single S locus with multiple alleles. In recent years, much progress has been made in determining the male and female S determinant in Brassica species. In the female, a gain-of function experiment clearly demonstrated that SRK was the sole S determinant, and that SLG enhanced the SI recognition process. By contrast, the male S determinant (termed SP11/SCR) was identified in the course of genome analysis of S locus to be a small cysteine-rich protein, which was classified as a pollen coat protein. This SP11/SCR may function as a ligand for the S domain of SRK in the SI recognition reaction of Brassica species. PMID- 11427675 TI - Ancestral MADS box genes in Sugi, Cryptomeria japonica D. Don (Taxodiaceae), homologous to the B function genes in angiosperms. AB - In flowering plants, flower organ identity is controlled by the ABC genes, including several MADS box genes. We present two MADS box genes of a conifer, Cryptomeria japonica D. Don. The genes, CjMADS1 and CjMADS2, were related to the angiosperm B function genes which determine the identities of petals and stamens. A phylogenetic analysis showed that these genes form a new clade outside the angiosperm B group, that is, PISTILLATA (PI) and APETALA3 (AP3) lineages. CjMADS1 had a PI-group specific motif and CjMADS2 had AP3-group specific motifs at the C terminal end, respectively. CjMADS1 was expressed in male strobili (or cones) throughout its development, while CjMADS2 was transiently expressed during male strobilus development. The specific expression in the male reproductive organ indicated that the B function is maintained in gymnosperms. Our cladistic analysis suggests that the gene duplication event which generated B function gene lineages predates the divergence of angiosperms and gymnosperms and that the gene duplication which produced the two genes of C. japonica occurred in an ancestral conifer species. PMID- 11427676 TI - Characterization of cDNA of the liverwort phytochrome gene, and phytochrome involvement in the light-dependent and light-independent protochlorophyllide oxidoreductase gene expression in Marchantia paleacea var. diptera. AB - The cDNA of the phytochrome gene in the liverwort Marchantia paleacea var. diptera (MpdPHY1) was isolated. MpdPHY1 encoded a conventional phytochrome apoprotein. The MpdPHY1 transcript was accumulated in the dark and suppressed in the light. The degradation of the MpdPHY1 transcript by red light irradiation had red/far-red reversibility, suggesting that the liverwort phytochrome gene expression was regulated by a phytochrome. Northern blot analysis of the transcripts in cells irradiated by red/far-red light revealed that the liverwort phytochrome was involved in the expressions of chlB, chlL, chlN, or por, which encode subunits of light-independent and light-dependent protochlorophyllide oxidoreductase, respectively. PMID- 11427677 TI - Spatial and temporal distribution of sucrose synthase in the radish hypocotyl in relation to thickening growth. AB - Sucrose synthase (SuSy) is a key enzyme in the development of storage root of radish. Clarification of its spatial and temporal expression during the thickening growth of radish hypocotyl, which later develops into storage root, was carried out immunologically using light microscopy. Sequential harvests at 3, 7, 11 and 13 d after sowing (DAS) were performed on two radish cultivars having different sink capacity. A very low level of SuSy was observed 3 DAS for both cultivars. White Cherrish (WC; strong storage root) showed the maximum level of SuSy between 7 and 11 DAS with increased cell development (thickening), while in Kosena (K; low storage root) the level remained high after 13 d of growth. A high level of SuSy was found in companion cells, which was consistent with previous observations, but SuSy was also found in the xylem parenchyma and in some cortical cells. The level of SuSy differed according to the localization and depended highly on cell development. Both cell division and cell enlargement were stimulated in WC compared with K. The role of SuSy during thickening growth of radish hypocotyl is discussed in terms of utilizing photosynthates. PMID- 11427678 TI - Turgor pressure regulation and the orientation of cortical microtubules in Spirogyra cells. AB - Microtubules (MTs) of cells of Spirogyra sp. were depolymerized by treatment with amiprophos-methyl (APM) for 1 h and then reorganized in 0.30 M mannitol solution. The reorganized MTs after 1.5 h incubation showed an oblique/longitudinal orientation and then became transversely oriented as the incubation was prolonged. During this incubation, the osmotic pressure of cells was measured by the plasmolysis method. The cell osmotic pressure increased with time. The calculated turgor pressure at 1.5 h was 0.11 M (mannitol equivalent) and, at 13.5 h, 0.25 M. Similar changes in MT orientation and recovery of the turgor pressure were also observed in 0.30 M sorbitol solution. These results suggest that the MT orientation may be correlated with the turgor pressure. Among fresh water algae sensitive to a saline environment, this Spirogyra was the first species shown to have a turgor regulating mechanism, although the recovery of turgor pressure was incomplete. The recovery of turgor pressure in mannitol solutions was also observed without APM treatment. PMID- 11427679 TI - Functional analysis of psbV and a novel c-type cytochrome gene psbV2 of the thermophilic cyanobacterium Thermosynechococcus elongatus strain BP-1. AB - Cytochrome c-550 is an extrinsic protein associated with photosystem II (PSII) in cyanobacteria and lower eukaryotic algae and plays an important role in the water splitting reaction. The gene (psbV) for cytochrome c-550 was cloned from the thermophilic cyanobacteria Thermosynechococcus (formerly Synechococcus) elongatus and T. (formerly Synechococcus) vulcanus. In both genomes, located downstream of psbV were a novel gene (designated psbV2) for a c-type cytochrome and petJ for cytochrome c-553. The deduced product of psbV2 showed composite similarities to psbV and petJ. Phenotype of psbV-disruptant in Thermosynechococcus was practically the same as that reported in Synechocystis sp. PCC 6803. Either psbV or psbV2 gene of T. elongatus was expressed in the psbV-disruptant of Synechocystis sp. PCC 6803, which resulted in recovery of the photoautotrophic growth. However, the enhanced requirement of Ca(2+) or Cl- ions in the psbV disruptant of Synechocystis was suppressed by expression of psbV but not by expression of psbV2. Thus, it is concluded that psbV2 can partly replace the role of psbV in PSII. The close tandem arrangement of psbV/psbV2/petJ implies that psbV2 was created by gene duplication and intergenic recombination during evolution. PMID- 11427680 TI - The M locus and ethylene-controlled sex determination in andromonoecious cucumber plants. AB - Sex determination in cucumber (Cucumis sativus L.) plants is genetically controlled by the F and M loci. These loci interact to produce three different sexual phenotypes: gynoecious (M-F-), monoecious (M-ff), and andromonoecious (mmff). Gynoecious cucumber plants produce more ethylene than do monoecious plants. We found that the levels of ethylene production and the accumulation of CS-ACS2 mRNA in andromonoecious cucumber plants did not differ from those in monoecious plants and were lower than the levels measured in gynoecious plants. Ethylene inhibited stamen development in gynoecious cucumbers but not in andromonoecious ones. Furthermore, ethylene caused substantial increases in the accumulation of CS-ETR2, CS-ERS, and CS-ACS2 mRNA in monoecious and gynoecious cucumber plants, but not in andromonoecious one. In addition, the inhibitory effect of ethylene on hypocotyl elongation in andromonoecious cucumber plants was less than that in monoecious and gynoecious plants. These results suggest that ethylene responses in andromonoecious cucumber plants are reduced from those in monoecious and gynoecious plants. This is the first evidence that ethylene signals may influence the product of the M locus and thus inhibit stamen development in cucumber. The andromonoecious line provides novel material for studying the function of the M locus during sex determination in flowering cucumbers. PMID- 11427681 TI - Different cytoplasmic calcium contents among three species of Characeae. AB - Internodal cells of three species of Characeae, Nitella flexilis, Nitella axilliformis and Chara corallina, were analyzed for the contents of Ca(2+ )and Mg(2+) in the cytoplasm. To avoid contamination of Ca(2+) from the cell wall and vacuole, the vacuolar sap was replaced with a sorbitol solution containing Sr(2+) by the vacuolar perfusion method after the cell had been treated with Sr(2+). No significant difference in the cytoplasmic content of Mg(2+) was found among three species of Characeae, but significant differences in the cytoplasmic content of Ca(2+) were observed among them. The cytoplasmic Ca(2+) content of N. flexilis was 2.0 times that of N. axilliformis and 3.3 times that of C. corallina. The cytoplasmic drop was furthermore separated into two fractions: a chloroplast-free fraction and a chloroplast fraction. In the chloroplast-free fraction the Ca(2+) content of N. flexilis was 2.3 times that of C. corallina and 2.0 times that of N. axilliformis, while the Mg(2+) content was the same among the three species. In the chloroplast fraction N. flexilis contained about seven times more Ca(2+) and about two times more Mg(2+) than C. corallina. The difference in the cytoplasmic Ca(2+ )content was discussed in relation to the difference in the capacity for the hydration-induced Ca(2+) release existing among the three species. PMID- 11427682 TI - Serine acetyltransferase involved in cysteine biosynthesis from spinach: molecular cloning, characterization and expression analysis of cDNA encoding a plastidic isoform. AB - A cDNA clone that encodes a chloroplast-localizing isoform of serine acetyltransferase (SATase) (EC 2.3.1.30) was isolated from spinach (Spinacia oleracea L.). The cDNA encodes a polypeptide of 347 amino acids containing a putative transit peptide of ca. 60-70 amino acids at the N-terminal. Deduced amino acid sequence of SATase from spinach exhibited homology with other SATases from plants. DNA blot hybridization analysis showed the presence of 2-3 copies of Sat gene in the genome of spinach. RNA blot hybridization analysis indicated the constitutive expression of Sat gene in green and etiolated seedlings of spinach. Bacterial expression of the cDNA could directly rescue the cysteine auxotrophy of Escherchia coli caused by a lack of SATase locus (cysE). Catalytically active SATase protein was produced in E. coli cells. L-Cysteine, an end product of the cysteine biosynthetic pathway, inhibited the activity of recombinant spinach SATase, indicating the regulatory function of SATase in this metabolic pathway. A chloroplastic localization of this spinach SATase was revealed by the analyses of transgenic plant expressing transit peptide of SATase-beta-glucuronidase (GUS) fusion protein, and transient expression using the transit peptide-green fluorescent protein (GFP) fusion protein. The result from in vitro translation analysis suggests that this cDNA may encode both plastidic and cytosolic SATases. PMID- 11427683 TI - A TGACGT motif in the 5'-upstream region of alpha-amylase gene from Vigna mungo is a cis-element for expression in cotyledons of germinated seeds. AB - Alpha-amylase is expressed at high levels in cotyledons of germinated seeds of Vigna mungo. The mRNA for alpha-amylase appeared in cotyledons of the seeds at 1 d after imbibition started (DAI). Two TGACGT motifs at -445 and at -125 in the promoter region of the gene interacted with nuclear proteins from cotyledons of dry seeds and the activities were detected until 3 DAI. A transient assay with particle bombardment showed that the downstream region from -135 in the promoter was required for high level expression in the cotyledons and the activity was reduced by mutation of the TGACGT motif at -125. The activities to bind the TGACGT motifs were detected in the axes of the seeds at 1 DAI but disappeared at 4 DAI, although the mRNA for alpha-amylase in the axes appeared at 4 DAI and increased in level by 6 DAI. A transient assay experiment showed that a positive regulatory element for the expression in the axes was located in the region from 630 to -453. These results indicated that the TGACGT motif at -125 was required for high level expression of the gene in the cotyledons of the germinated seeds. PMID- 11427684 TI - Protein phosphorylation in pea root plastids. AB - Protein phosphorylation has been investigated in non-photosynthetic plastids of pea roots. Intact and lysed preparations of plastids were incubated with [gamma (32)P]ATP and three stromal proteins of sizes 41, 58 and 62 kDa were phosphorylated on a serine residue. No other proteins were significantly labelled under the conditions used. The 62 kDa protein is probably phosphoglucomutase and represents a phosphoenzyme catalytic intermediate. The protein kinase(s) and phosphatase(s) acting on the other proteins were not sensitive to exogenous calcium but were sensitive to magnesium. The protein phosphatase which acts on the 41 kDa protein is possibly of type 2C, whereas that acting on the 58 kDa phosphoprotein did not fall into any class defined by mammalian systems. Metabolism of exogenous glucose 6-phosphate by the oxidative pentose phosphate pathway in intact plastids abolished the phosphorylation of the 58 kDa protein. Dihydroxyacetone phosphate, phosphoenolpyruvate and 3-phosphoglycerate also inhibited phosphorylation of the 58 kDa protein and had a time-dependent effect on the phosphorylation of the 41 kDa protein. The significance of these results in relation to a possible role for protein phosphorylation in these plastids is considered. PMID- 11427685 TI - Lipoic acid metabolism in Arabidopsis thaliana: cloning and characterization of a cDNA encoding lipoyltransferase. AB - Lipoic acid is an essential coenzyme required for activity of several key enzyme complexes, such as the pyruvate dehydrogenase complex, in the central metabolism. In these complexes, lipoic acid must be covalently attached to one of the component proteins for it to have biological activity. We report the cloning and characterization of Arabidopsis thaliana LIP2 cDNA for lipoyltransferase that catalyzes the transfer of the lipoyl group from lipoyl-acyl carrier protein to lipoate-dependent enzymes. This cDNA was shown to code for lipoyltransferase by its ability to complement an Escherichia coli lipB null mutant lacking lipoyltransferase activity. The expressed enzyme in the E. coli mutant efficiently complemented the activity of pyruvate dehydrogenase complex, but less efficiently than that of 2-oxoglutarate dehydrogenase complex. Comparison of the deduced amino acid sequence of LIP2 with those of E. coli and yeast lipoyltransferases showed a marked sequence similarity and the presence of a leader sequence presumably required for import into mitochondria. Southern and northern hybridization analyses suggest that LIP2 is a single-copy gene and is expressed as an mRNA of 860 nt in leaves. Western blot analysis with an antibody against lipoyltransferase demonstrated that a 29 kDa form of lipoyltransferase is located in the mitochondrial compartment of A. thaliana. PMID- 11427686 TI - Growth in microgravity increases susceptibility of soybean to a fungal pathogen. AB - The influence of microgravity on the susceptibility of soybean roots to Phytophthora sojae was studied during the Space Shuttle Mission STS-87. Seedlings of soybean cultivar Williams 82 grown in spaceflight or at unit gravity were untreated or inoculated with the soybean root rot pathogen P. sojae. At 3, 6 and 7 d after launch while still in microgravity, seedlings were photographed and then fixed for subsequent microscopic analysis. Post-landing analysis of the seedlings revealed that at harvest day 7 the length of untreated roots did not differ between flight and ground samples. However, the flight-grown roots infected with P. sojae showed more disease symptoms (percentage of brown and macerated areas) and the root tissues were more extensively colonized relative to the ground controls exposed to the fungus. Ethylene levels were higher in spaceflight when compared to ground samples. These data suggest that soybean seedlings grown in microgravity are more susceptible to colonization by a fungal pathogen relative to ground controls. PMID- 11427687 TI - The sensitivity of chlamydomonas photoreceptor is optimized for the frequency of cell body rotation. AB - For phototactic migration, Chlamydomonas scans the surrounding light environment by rotating the cell body with an eyespot located on the equator. The intensity of the light signal received by the eyespot should therefore change cyclically at the frequency of the cell body rotation. In this study, the response of the photoreceptor to cyclically changing light stimuli was analyzed using immotile mutant cells. To simulate the light intensity change perceived by a rotating cell, light stimuli were applied that consisted of a light phase with the intensity changing similar to a half cycle of a sine wave and a dark phase of the same length. The fluence rate at the peak of the sine wave was of the order of 10(19) photons m(-2) s(-1), i.e. high intensity at which phototaxis is saturated. A photoreceptor current (PRC) was produced at the onset of each light phase. Interestingly, its amplitude varied depending on the frequency and was largest at 1-5 Hz, a frequency range similar to the frequency of cell body rotation. Experiments on the kinetics of the PRC indicate that the response was small at low frequency because of the inactivation of the PRC before full activation. In contrast, at high frequency the PRC was suppressed by adaptation to the repetitive stimuli. These characteristic kinetics of the PRC should be important for Chlamydomonas cells to extract information from the signals generated by the cell body rotation. PMID- 11427688 TI - A possible role of glutathione and glutathione disulfide in tracheary element differentiation in the cultured mesophyll cells of Zinnia elegans. AB - We investigated the relationship between the cellular redox state of GSH or GSSG and tracheary element (TE) differentiation using a Zinnia experimental system, in which isolated mesophyll cells transdifferentiate to TEs. TE differentiation was suppressed by the application of L-buthionine sulfoximine (BSO), a potent inhibitor of GSH biosynthesis, at the early stage of cell culture. Application of GSSG at the early culture stage promoted the differentiation, but that of GSH or GSSG at an advanced period of culture suppressed the differentiation. Application of GSH and GSSG nullified the TE differentiation-suppressing effect of BSO. The results suggest that changes in the redox states of GSH and GSSG have a role in TE differentiation. PMID- 11427689 TI - Molecular physiology of kainate receptors. AB - A decade ago, our understanding of the molecular properties of kainate receptors and their involvement in synaptic physiology was essentially null. A plethora of recent studies has altered this situation profoundly such that kainate receptors are now regarded as key players in the modulation of transmitter release, as important mediators of the postsynaptic actions of glutamate, and as possible targets for the development of antiepileptic and analgesic drugs. In this review, we summarize our current knowledge of the properties of kainate receptors focusing on four key issues: 1) their structural and biophysical features, 2) the important progress in their pharmacological characterization, 3) their pre- and postsynaptic mechanisms of action, and 4) their involvement in a series of physiological and pathological processes. Finally, although significant progress has been made toward the elucidation of their importance for brain function, kainate receptors remain largely an enigma and, therefore, we propose some new roads that should be explored to obtain a deeper understanding of this young, but intriguing, class of proteins. PMID- 11427690 TI - Vascular smooth muscle growth: autocrine growth mechanisms. AB - Vascular smooth muscle cells (VSMC) exhibit several growth responses to agonists that regulate their function including proliferation (hyperplasia with an increase in cell number), hypertrophy (an increase in cell size without change in DNA content), endoreduplication (an increase in DNA content and usually size), and apoptosis. Both autocrine growth mechanisms (in which the individual cell synthesizes and/or secretes a substance that stimulates that same cell type to undergo a growth response) and paracrine growth mechanisms (in which the individual cells responding to the growth factor synthesize and/or secrete a substance that stimulates neighboring cells of another cell type) are important in VSMC growth. In this review I discuss the autocrine and paracrine growth factors important for VSMC growth in culture and in vessels. Four mechanisms by which individual agonists signal are described: direct effects of agonists on their receptors, transactivation of tyrosine kinase-coupled receptors, generation of reactive oxygen species, and induction/secretion of other growth and survival factors. Additional growth effects mediated by changes in cell matrix are discussed. The temporal and spatial coordination of these events are shown to modulate the environment in which other growth factors initiate cell cycle events. Finally, the heterogeneous nature of VSMC developmental origin provides another level of complexity in VSMC growth mechanisms. PMID- 11427691 TI - Short-chain fatty acids and human colonic function: roles of resistant starch and nonstarch polysaccharides. AB - Resistant starch (RS) is starch and products of its small intestinal digestion that enter the large bowel. It occurs for various reasons including chemical structure, cooking of food, chemical modification, and food mastication. Human colonic bacteria ferment RS and nonstarch polysaccharides (NSP; major components of dietary fiber) to short-chain fatty acids (SCFA), mainly acetate, propionate, and butyrate. SCFA stimulate colonic blood flow and fluid and electrolyte uptake. Butyrate is a preferred substrate for colonocytes and appears to promote a normal phenotype in these cells. Fermentation of some RS types favors butyrate production. Measurement of colonic fermentation in humans is difficult, and indirect measures (e.g., fecal samples) or animal models have been used. Of the latter, rodents appear to be of limited value, and pigs or dogs are preferable. RS is less effective than NSP in stool bulking, but epidemiological data suggest that it is more protective against colorectal cancer, possibly via butyrate. RS is a prebiotic, but knowledge of its other interactions with the microflora is limited. The contribution of RS to fermentation and colonic physiology seems to be greater than that of NSP. However, the lack of a generally accepted analytical procedure that accommodates the major influences on RS means this is yet to be established. PMID- 11427692 TI - Mechanisms of spreading depression and hypoxic spreading depression-like depolarization. AB - Spreading depression (SD) and the related hypoxic SD-like depolarization (HSD) are characterized by rapid and nearly complete depolarization of a sizable population of brain cells with massive redistribution of ions between intracellular and extracellular compartments, that evolves as a regenerative, "all-or-none" type process, and propagates slowly as a wave in brain tissue. This article reviews the characteristics of SD and HSD and the main hypotheses that have been proposed to explain them. Both SD and HSD are composites of concurrent processes. Antagonists of N-methyl-D-aspartate (NMDA) channels or voltage-gated Na(+) or certain types of Ca(2+) channels can postpone or mitigate SD or HSD, but it takes a combination of drugs blocking all known major inward currents to effectively prevent HSD. Recent computer simulation confirmed that SD can be produced by positive feedback achieved by increase of extracellular K(+) concentration that activates persistent inward currents which then activate K(+) channels and release more K(+). Any slowly inactivating voltage and/or K(+) dependent inward current could generate SD-like depolarization, but ordinarily, it is brought about by the cooperative action of the persistent Na(+) current I(Na,P) plus NMDA receptor-controlled current. SD is ignited when the sum of persistent inward currents exceeds persistent outward currents so that total membrane current turns inward. The degree of depolarization is not determined by the number of channels available, but by the feedback that governs the SD process. Short bouts of SD and HSD are well tolerated, but prolonged depolarization results in lasting loss of neuron function. Irreversible damage can, however, be avoided if Ca(2+) influx into neurons is prevented. PMID- 11427693 TI - Physiological and molecular basis of thyroid hormone action. AB - Thyroid hormones (THs) play critical roles in the differentiation, growth, metabolism, and physiological function of virtually all tissues. TH binds to receptors that are ligand-regulatable transcription factors belonging to the nuclear hormone receptor superfamily. Tremendous progress has been made recently in our understanding of the molecular mechanisms that underlie TH action. In this review, we present the major advances in our knowledge of the molecular mechanisms of TH action and their implications for TH action in specific tissues, resistance to thyroid hormone syndrome, and genetically engineered mouse models. PMID- 11427694 TI - Cerebellar long-term depression: characterization, signal transduction, and functional roles. AB - Cerebellar Purkinje cells exhibit a unique type of synaptic plasticity, namely, long-term depression (LTD). When two inputs to a Purkinje cell, one from a climbing fiber and the other from a set of granule cell axons, are repeatedly associated, the input efficacy of the granule cell axons in exciting the Purkinje cell is persistently depressed. Section I of this review briefly describes the history of research around LTD, and section II specifies physiological characteristics of LTD. Sections III and IV then review the massive data accumulated during the past two decades, which have revealed complex networks of signal transduction underlying LTD. Section III deals with a variety of first messengers, receptors, ion channels, transporters, G proteins, and phospholipases. Section IV covers second messengers, protein kinases, phosphatases and other elements, eventually leading to inactivation of DL-alpha amino-3-hydroxy-5-methyl-4-isoxazolone-propionate-selective glutamate receptors that mediate granule cell-to-Purkinje cell transmission. Section V defines roles of LTD in the light of the microcomplex concept of the cerebellum as functionally eliminating those synaptic connections associated with errors during repeated exercises, while preserving other connections leading to the successful execution of movements. Section VI examines the validity of this microcomplex concept based on the data collected from recent numerous studies of various forms of motor learning in ocular reflexes, eye-blink conditioning, posture, locomotion, and hand/arm movements. Section VII emphasizes the importance of integrating studies on LTD and learning and raises future possibilities of extending cerebellar research to reveal memory mechanisms of implicit learning in general. PMID- 11427695 TI - Gene regulation in the magnocellular hypothalamo-neurohypophysial system. AB - The hypothalamo-neurohypophysial system (HNS) is the major peptidergic neurosecretory system through which the brain controls peripheral physiology. The hormones vasopressin and oxytocin released from the HNS at the neurohypophysis serve homeostatic functions of water balance and reproduction. From a physiological viewpoint, the core question on the HNS has always been, "How is the rate of hormone production controlled?" Despite a clear description of the physiology, anatomy, cell biology, and biochemistry of the HNS gained over the last 100 years, this question has remained largely unanswered. However, recently, significant progress has been made through studies of gene identity and gene expression in the magnocellular neurons (MCNs) that constitute the HNS. These are keys to mechanisms and events that exist in the HNS. This review is an inventory of what we know about genes expressed in the HNS, about the regulation of their expression in response to physiological stimuli, and about their function. Genes relevant to the central question include receptors and signal transduction components that receive and process the message that the organism is in demand of a neurohypophysial hormone. The key players in gene regulatory events, the transcription factors, deserve special attention. They do not only control rates of hormone production at the level of the gene, but also determine the molecular make-up of the cell essential for appropriate development and physiological functioning. Finally, the HNS neurons are equipped with a machinery to produce and secrete hormones in a regulated manner. With the availability of several gene transfer approaches applicable to the HNS, it is anticipated that new insights will be obtained on how the HNS is able to respond to the physiological demands for its hormones. PMID- 11427696 TI - Nuclear hormone receptors and gene expression. AB - The nuclear hormone receptor superfamily includes receptors for thyroid and steroid hormones, retinoids and vitamin D, as well as different "orphan" receptors of unknown ligand. Ligands for some of these receptors have been recently identified, showing that products of lipid metabolism such as fatty acids, prostaglandins, or cholesterol derivatives can regulate gene expression by binding to nuclear receptors. Nuclear receptors act as ligand-inducible transcription factors by directly interacting as monomers, homodimers, or heterodimers with the retinoid X receptor with DNA response elements of target genes, as well as by "cross-talking" to other signaling pathways. The effects of nuclear receptors on transcription are mediated through recruitment of coregulators. A subset of receptors binds corepressor factors and actively represses target gene expression in the absence of ligand. Corepressors are found within multicomponent complexes that contain histone deacetylase activity. Deacetylation leads to chromatin compactation and transcriptional repression. Upon ligand binding, the receptors undergo a conformational change that allows the recruitment of multiple coactivator complexes. Some of these proteins are chromatin remodeling factors or possess histone acetylase activity, whereas others may interact directly with the basic transcriptional machinery. Recruitment of coactivator complexes to the target promoter causes chromatin decompactation and transcriptional activation. The characterization of corepressor and coactivator complexes, in concert with the identification of the specific interaction motifs in the receptors, has demonstrated the existence of a general molecular mechanism by which different receptors elicit their transcriptional responses in target genes. PMID- 11427698 TI - Spectrin and ankyrin-based pathways: metazoan inventions for integrating cells into tissues. AB - The spectrin-based membrane skeleton of the humble mammalian erythrocyte has provided biologists with a set of interacting proteins with diverse roles in organization and survival of cells in metazoan organisms. This review deals with the molecular physiology of spectrin, ankyrin, which links spectrin to the anion exchanger, and two spectrin-associated proteins that promote spectrin interactions with actin: adducin and protein 4.1. The lack of essential functions for these proteins in generic cells grown in culture and the absence of their genes in the yeast genome have, until recently, limited advances in understanding their roles outside of erythrocytes. However, completion of the genomes of simple metazoans and application of homologous recombination in mice now are providing the first glimpses of the full scope of physiological roles for spectrin, ankyrin, and their associated proteins. These functions now include targeting of ion channels and cell adhesion molecules to specialized compartments within the plasma membrane and endoplasmic reticulum of striated muscle and the nervous system, mechanical stabilization at the tissue level based on transcellular protein assemblies, participation in epithelial morphogenesis, and orientation of mitotic spindles in asymmetric cell divisions. These studies, in addition to stretching the erythrocyte paradigm beyond recognition, also are revealing novel cellular pathways essential for metazoan life. Examples are ankyrin-dependent targeting of proteins to excitable membrane domains in the plasma membrane and the Ca(2+) homeostasis compartment of the endoplasmic reticulum. Exciting questions for the future relate to the molecular basis for these pathways and their roles in a clinical context, either as the basis for disease or more positively as therapeutic targets. PMID- 11427697 TI - Mechanics of the mammalian cochlea. AB - In mammals, environmental sounds stimulate the auditory receptor, the cochlea, via vibrations of the stapes, the innermost of the middle ear ossicles. These vibrations produce displacement waves that travel on the elongated and spirally wound basilar membrane (BM). As they travel, waves grow in amplitude, reaching a maximum and then dying out. The location of maximum BM motion is a function of stimulus frequency, with high-frequency waves being localized to the "base" of the cochlea (near the stapes) and low-frequency waves approaching the "apex" of the cochlea. Thus each cochlear site has a characteristic frequency (CF), to which it responds maximally. BM vibrations produce motion of hair cell stereocilia, which gates stereociliar transduction channels leading to the generation of hair cell receptor potentials and the excitation of afferent auditory nerve fibers. At the base of the cochlea, BM motion exhibits a CF specific and level-dependent compressive nonlinearity such that responses to low level, near-CF stimuli are sensitive and sharply frequency-tuned and responses to intense stimuli are insensitive and poorly tuned. The high sensitivity and sharp frequency tuning, as well as compression and other nonlinearities (two-tone suppression and intermodulation distortion), are highly labile, indicating the presence in normal cochleae of a positive feedback from the organ of Corti, the "cochlear amplifier." This mechanism involves forces generated by the outer hair cells and controlled, directly or indirectly, by their transduction currents. At the apex of the cochlea, nonlinearities appear to be less prominent than at the base, perhaps implying that the cochlear amplifier plays a lesser role in determining apical mechanical responses to sound. Whether at the base or the apex, the properties of BM vibration adequately account for most frequency specific properties of the responses to sound of auditory nerve fibers. PMID- 11427699 TI - Essential roles for four cytoplasmic intermediate filament proteins in Caenorhabditis elegans development. AB - The structural proteins of the cytoplasmic intermediate filaments (IFs) arise in the nematode Caenorhabditis elegans from eight reported genes and an additional three genes now identified in the complete genome. With the use of double stranded RNA interference (RNAi) for all 11 C. elegans genes encoding cytoplasmic IF proteins, we observe phenotypes for the five genes A1, A2, A3, B1, and C2. These range from embryonic lethality (B1) and embryonic/larval lethality (A3) to larval lethality (A1 and A2) and a mild dumpy phenotype of adults (C2). Phenotypes A2 and A3 involve displaced body muscles and paralysis. They probably arise by reduction of hypodermal IFs that participate in the transmission of force from the muscle cells to the cuticle. The B1 phenotype has multiple morphogenetic defects, and the A1 phenotype is arrested at the L1 stage. Thus, at least four IF genes are essential for C. elegans development. Their RNAi phenotypes are lethal defects due to silencing of single IF genes. In contrast to C. elegans, no IF genes have been identified in the complete Drosophila genome, posing the question of how Drosophila can compensate for the lack of these proteins, which are essential in mammals and C. elegans. We speculate that the lack of IF proteins in Drosophila can be viewed as cytoskeletal alteration in which, for instance, stable microtubules, often arranged as bundles, substitute for cytoplasmic IFs. PMID- 11427700 TI - Prospective measurements of dehydroepiandrosterone sulfate in a cohort of elderly subjects: relationship to gender, subjective health, smoking habits, and 10-year mortality. AB - The decrease with age of the adrenal-secreted dehydroepiandrosterone sulfate (DHEAS) in serum has suggested that it may be causally related to longevity. For the PAQUID [People (Personnes) Aged (Agees) About What (Quid, in Latin)] cohort of elderly subjects, we have previously reported higher DHEAS in men than in women, a decrease with age and, among men, a negative correlation between the DHEAS level and mortality at 2 and 4 years. Here, with an 8-year followup in 290 subjects, we show a global decrease of 2.3% per year for men and 3.9% per year for women. However, in approximately 30% of cases, there was an increase of DHEAS. We observed no relationship between the evolution of DHEAS level and functional, psychological, and mental status, possibly because of selection by death. In women, no association was found between mortality and DHEAS level. In men, the relative risk (RR) of death was higher for the lowest levels of DHEAS (RR = 1.9, P = 0.007), with RR = 6.5, P = 0.003 for those under 70 years old, a result indicating heterogeneity of the population. There was an effect of subjective health on mortality that disappeared after adjustment of DHEAS levels, suggesting its relation with these DHEAS levels. Death RR was much higher in smokers with a low DHEAS level than in nonsmokers with high DHEAS (RR = 6.7, P = 0.001). We submit that the involvement of DHEAS is possibly different according to gender, that association between low DHEAS level and mortality only for men under 70 years old possibly reflects heterogeneity of the population, and that DHEAS level is a reliable predictor of death in male smokers. PMID- 11427701 TI - Excitatory versus inhibitory GABA as a divergence point in steroid-mediated sexual differentiation of the brain. AB - Whereas adult sex differences in brain morphology and behavior result from developmental exposure to steroid hormones, the mechanism by which steroids differentiate the brain is unknown. Studies to date have described subtle sex differences in levels of proteins and neurotransmitters during brain development, but these have lacked explanatory power for the profound sex differences induced by steroids. We report here a major divergence in the response to injection of the gamma-aminobutyric acid type A (GABA(A)) agonist, muscimol, in newborn male and female rats. In females, muscimol treatment primarily decreased the phosphorylation of cAMP response element binding protein (CREB) within the hypothalamus and the CA1 region of the hippocampus. In contrast, muscimol increased the phosphorylation of CREB in males within these same brain regions. Within the arcuate nucleus, muscimol treatment increased the phosphorylation of CREB in both females and males. Thus, the response to GABA can be excitatory or inhibitory on signal-transduction pathways that alter CREB phosphorylation depending on the sex and the region in developing brain. This divergence in response to GABA allows for a previously unknown form of steroid-mediated neuronal plasticity and may be an initial step in establishing sexually dimorphic signal-transduction pathways in developing brain. PMID- 11427702 TI - The origin of 15R-prostaglandins in the Caribbean coral Plexaura homomalla: molecular cloning and expression of a novel cyclooxygenase. AB - The highest concentrations of prostaglandins in nature are found in the Caribbean gorgonian Plexaura homomalla. Depending on its geographical location, this coral contains prostaglandins with typical mammalian stereochemistry (15S-hydroxy) or the unusual 15R-prostaglandins. Their metabolic origin has remained the subject of mechanistic speculations for three decades. Here, we report the structure of a type of cyclooxygenase (COX) that catalyzes transformation of arachidonic acid into 15R-prostaglandins. Using a homology-based reverse transcriptase--PCR strategy, we cloned a cDNA corresponding to a COX protein from the R variety of P. homomalla. The deduced peptide sequence shows 80% identity with the 15S specific coral COX from the Arctic soft coral Gersemia fruticosa and approximately 50% identity to mammalian COX-1 and COX-2. The predicted tertiary structure shows high homology with mammalian COX isozymes having all of the characteristic structural units and the amino acid residues important in catalysis. Some structural differences are apparent around the peroxidase active site, in the membrane-binding domain, and in the pattern of glycosylation. When expressed in Sf9 cells, the P. homomalla enzyme forms a 15R-prostaglandin endoperoxide together with 11R-hydroxyeicosatetraenoic acid and 15R hydroxyeicosatetraenoic acid as by-products. The endoperoxide gives rise to 15R prostaglandins and 12R-hydroxyheptadecatrienoic acid, identified by comparison to authentic standards. Evaluation of the structural differences of this 15R-COX isozyme should provide new insights into the substrate binding and stereospecificity of the dioxygenation reaction of arachidonic acid in the cyclooxygenase active site. PMID- 11427703 TI - Tsg101, a homologue of ubiquitin-conjugating (E2) enzymes, binds the L domain in HIV type 1 Pr55(Gag). AB - Ubiquitination appears to be involved in virus particle release from infected cells. Free ubiquitin (Ub), as well as Ub covalently bound to a small fraction of p6 Gag, is detected in mature HIV particles. Here we report that the p6 region in the Pr55(Gag) structural precursor polyprotein binds to Tsg101, a putative Ub regulator that is involved in trafficking of plasma membrane-associated proteins. Tsg101 was found to interact with Gag in (i) a yeast two-hybrid assay, (ii) in vitro coimmunoprecipitation by using purified Pr55(Gag) and rabbit reticulocyte lysate-synthesized Tsg101, and (iii) in vivo in the cytoplasm of COS cells transfected with gag. The PTAPP motif [or late (L) domain] within p6, which is required for release of mature virus from the plasma membrane, was the determinant for binding Pr55(Gag). The N-terminal region in Tsg101, which is homologous to the Ubc4 class of Ub-conjugating (E2) enzymes, was the determinant of interaction with p6. Mutation of Tyr-110 in Tsg101, present in place of the active-site Cys that binds Ub in E2 enzymes, and other residues unique to Tsg101, impaired p6 interaction, indicating that features that distinguish Tsg101 from active E2 enzymes were important for binding the viral protein. The results link L-domain function in HIV to the Ub machinery and a specific component of the cellular trafficking apparatus. PMID- 11427704 TI - Submucosal gland secretions in airways from cystic fibrosis patients have normal [Na(+)] and pH but elevated viscosity. AB - Fluid and macromolecule secretion by submucosal glands in mammalian airways is believed to be important in normal airway physiology and in the pathophysiology of cystic fibrosis (CF). An in situ fluorescence method was applied to measure the ionic composition and viscosity of freshly secreted fluid from airway glands. Fragments of human large airways obtained at the time of lung transplantation were mounted in a humidified perfusion chamber and the mucosal surface was covered by a thin layer of oil. Individual droplets of secreted fluid were microinjected with fluorescent indicators for measurement of [Na(+)], [Cl(-)], and pH by ratio imaging fluorescence microscopy and viscosity by fluorescence recovery after photobleaching. After carbachol stimulation, 0.1--0.5 microl of fluid accumulated in spherical droplets at gland orifices in approximately 3--5 min. In gland fluid from normal human airways, [Na(+)] was 94 +/- 8 mM, [Cl(-)] was 92 +/- 12 mM, and pH was 6.97 +/- 0.06 (SE, n = 7 humans, more than five glands studied per sample). Apparent fluid viscosity was 2.7 +/- 0.3-fold greater than that of saline. Neither [Na(+)] nor pH differed in gland fluid from CF airways, but viscosity was significantly elevated by approximately 2-fold compared to normal airways. These results represent the first direct measurements of ionic composition and viscosity in uncontaminated human gland secretions and indicate similar [Na(+)], [Cl(-)], and pH to that in the airway surface liquid. The elevated gland fluid viscosity in CF may be an important factor promoting bacterial colonization and airway disease. PMID- 11427705 TI - The generation of oscillations in networks of electrically coupled cells. AB - In several biological systems, the electrical coupling of nonoscillating cells generates synchronized membrane potential oscillations. Because the isolated cell is nonoscillating and electrical coupling tends to equalize the membrane potentials of the coupled cells, the mechanism underlying these oscillations is unclear. Here we present a dynamic mechanism by which the electrical coupling of identical nonoscillating cells can generate synchronous membrane potential oscillations. We demonstrate this mechanism by constructing a biologically feasible model of electrically coupled cells, characterized by an excitable membrane and calcium dynamics. We show that strong electrical coupling in this network generates multiple oscillatory states with different spatio-temporal patterns and discuss their possible role in the cooperative computations performed by the system. PMID- 11427706 TI - Reovirus reverse genetics: Incorporation of the CAT gene into the reovirus genome. AB - We have modified the infectious reovirus RNA system so as to generate a reovirus reverse genetics system. The system consists of (i) the plus strands of nine wild type reovirus genome segments; (ii) transcripts of the genetically modified cDNA form of the tenth genome segment; and (iii) a cell line transformed so as to express the protein normally encoded by the tenth genome segment. In the work described here, we have generated a serotype 3 reovirus into the S2 double stranded RNA genome segment of which the CAT gene has been cloned. The virus is stable, replicates in cells that have been transformed (so as to express the S2 gene product, protein final sigma 2), and expresses high levels of CAT activity. This technology can be extended to members of the orbivirus and rotavirus genera. This technology provides a powerful system for basic studies of double-stranded RNA virus replication; a nonpathogenic viral vector that replicates to high titers and could be used for clinical applications; and a system for providing nonselectable viral variants (the result of mutations, insertions, and deletions) that could be valuable for the construction of viral vaccine strains against human and animal pathogens. PMID- 11427707 TI - Extreme morphological and ecological homoplasy in tropical salamanders. AB - Fossorial salamanders typically have elongate and attenuated heads and bodies, diminutive limbs, hands and feet, and extremely elongate tails. Batrachoseps from California, Lineatriton from eastern Mexico, and Oedipina from southern Mexico to Ecuador, all members of the family Plethodontidae, tribe Bolitoglossini, resemble one another in external morphology, which has evolved independently. Whereas Oedipina and Batrachoseps are elongate because there are more trunk vertebrae, a widespread homoplasy (parallelism) in salamanders, the genus Lineatriton is unique in having evolved convergently by an alternate "giraffe-neck" developmental program. Lineatriton has the same number of trunk vertebrae as related, nonelongated taxa, but individual trunk vertebrae are elongated. A robust phylogenetic hypothesis, based on sequences of three mtDNA genes, finds Lineatriton to be deeply nested within a clade characterized by generalized ecology and morphology. Lineatriton lineolus, the only currently recognized taxon in the genus, shows unanticipated genetic diversity. Surprisingly, geographically separated populations of L. lineolus are not monophyletic, but are sister taxa of different species of the morphologically generalized genus Pseudoeurycea. Lineatriton, long thought to be a unique monospecific lineage, is polyphyletic. Accordingly, the specialized morphology of Lineatriton displays homoplasy at two hierarchical levels: (i) with respect to other elongate lineages in the family (convergence), and (ii) within what is currently recognized as a single taxon (parallelism). These evolutionary events are of adaptive significance because to invade the lowland tropics salamanders must be either arboreal or fossorial; the repeated evolution of elongation and attenuation has led to multiple lowland invasions. PMID- 11427708 TI - A growth-constrained environment drives tumor progression invivo. AB - We recently have shown that selective growth of transplanted normal hepatocytes can be achieved in a setting of cell cycle block of endogenous parenchymal cells. Thus, massive proliferation of donor-derived normal hepatocytes was observed in the liver of rats previously given retrorsine (RS), a naturally occurring alkaloid that blocks proliferation of resident liver cells. In the present study, the fate of nodular hepatocytes transplanted into RS-treated or normal syngeneic recipients was followed. The dipeptidyl peptidase type IV-deficient (DPPIV(-)) rat model for hepatocyte transplantation was used to distinguish donor-derived cells from recipient cells. Hepatocyte nodules were chemically induced in Fischer 344, DPPIV(+) rats; livers were then perfused and larger (>5 mm) nodules were separated from surrounding tissue. Cells isolated from either tissue were then injected into normal or RS-treated DPPIV(-) recipients. One month after transplantation, grossly visible nodules (2--3 mm) were seen in RS-treated recipients transplanted with nodular cells. They grew rapidly, occupying 80--90% of the host liver at 2 months, and progressed to hepatocellular carcinoma within 4 months. By contrast, no liver nodules developed within 6 months when nodular hepatocytes were injected into the liver of recipients not exposed to RS, although small clusters of donor-derived cells were present in these animals. Taken together, these results directly point to a fundamental role played by the host environment in modulating the growth and the progression rate of altered cells during carcinogenesis. In particular, they indicate that conditions associated with growth constraint of the host tissue can drive tumor progression in vivo. PMID- 11427709 TI - Three SNARE complexes cooperate to mediate membrane fusion. AB - Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins of the syntaxin, SNAP-25, and VAMP families mediate intracellular membrane fusion through the formation of helical bundles that span opposing membranes. Soluble SNARE domains that lack their integral membrane anchors inhibit membrane fusion by forming nonfunctional complexes with endogenous SNARE proteins. In this study we investigate the dependence of membrane fusion on the concentration of a soluble SNARE coil domain derived from VAMP2. The increase in the inhibition of fusion observed with increasing concentration of inhibitor is best fit to a function that suggests three SNARE complexes cooperate to mediate fusion of a single vesicle. These three complexes likely contribute part of a protein and lipidic fusion pore. PMID- 11427710 TI - Life in the end-Permian dead zone. AB - The fossil record of land plants is an obvious source of information on the dynamics of mass extinctions in the geological past. In conjunction with the end Permian ecological crisis, approximately 250 million years ago, palynological data from East Greenland reveal some unanticipated patterns. We document the significant time lag between terrestrial ecosystem collapse and selective extinction among characteristic Late Permian plants. Furthermore, ecological crisis resulted in an initial increase in plant diversity, instead of a decrease. Paradoxically, these floral patterns correspond to a "dead zone" in the end Permian faunal record, characterized by a paucity of marine invertebrate megafossils. The time-delayed, end-Permian plant extinctions resemble modeled "extinction debt" responses of multispecies metapopulations to progressive habitat destruction. PMID- 11427711 TI - Wheat leaves emit nitrous oxide during nitrate assimilation. AB - Nitrous oxide (N(2)O) is a key atmospheric greenhouse gas that contributes to global climatic change through radiative warming and depletion of stratospheric ozone. In this report, N(2)O flux was monitored simultaneously with photosynthetic CO(2) and O(2) exchanges from intact canopies of 12 wheat seedlings. The rates of N(2)O-N emitted ranged from <2 pmol x m(-2) x s(-1) when NH(4)(+) was the N source, to 25.6 +/- 1.7 pmol x m(-2) x s(-1) (mean +/- SE, n = 13) when the N source was shifted to NO(3)(-). Such fluxes are among the smallest reported for any trace gas emitted by a higher plant. Leaf N(2)O emissions were correlated with leaf nitrate assimilation activity, as measured by using the assimilation quotient, the ratio of CO(2) assimilated to O(2) evolved. (15)N isotopic signatures on N(2)O emitted from leaves supported direct N(2)O production by plant NO(3)(-) assimilation and not N(2)O produced by microorganisms on root surfaces and emitted in the transpiration stream. In vitro production of N(2)O by both intact chloroplasts and nitrite reductase, but not by nitrate reductase, indicated that N(2)O produced by leaves occurred during photoassimilation of NO(2)(-) in the chloroplast. Given the large quantities of NO(3)(-) assimilated by plants in the terrestrial biosphere, these observations suggest that formation of N(2)O during NO(2)(-) photoassimilation could be an important global biogenic N(2)O source. PMID- 11427712 TI - A unique phospholipid organization in bovine erythrocyte membranes. AB - Ruminant erythrocytes are remarkable for their choline-phospholipid anomalies; namely, low or absent phosphatidylcholine (PC) along with high sphingomyelin levels. Here, we report another anomaly in bovine erythrocytes that affects aminophospholipids: phosphatidylethanolamine (PE) shows an extreme asymmetry, with only 2% of the total present in the outer leaflet. Furthermore, we found that phospholipase A(2), an enzyme located on the external surface of the erythrocytes, shows higher activity against PC than against PE. In addition, we observed that acylation of PE is by far the most important biosynthetic event in this system. We propose that deacylation of PE and PC by phospholipase A(2) to generate lysocompounds, followed by selective reacylation of lyso-PE in the inner leaflet, can account for the compositional and architectural peculiarities of bovine erythrocyte membranes. PMID- 11427713 TI - A TRP homolog in Saccharomyces cerevisiae forms an intracellular Ca(2+)-permeable channel in the yeast vacuolar membrane. AB - The molecular identification of ion channels in internal membranes has made scant progress compared with the study of plasma membrane ion channels. We investigated a prominent voltage-dependent, cation-selective, and calcium-activated vacuolar ion conductance of 320 pS (yeast vacuolar conductance, YVC1) in Saccharomyces cerevisiae. Here we report on a gene, the deduced product of which possesses significant homology to the ion channel of the transient receptor potential (TRP) family. By using a combination of gene deletion and re-expression with direct patch clamping of the yeast vacuolar membrane, we show that this yeast TRP-like gene is necessary for the YVC1 conductance. In physiological conditions, tens of micromolar cytoplasmic Ca(2+) activates the YVC1 current carried by cations including Ca(2+) across the vacuolar membrane. Immunodetection of a tagged YVC1 gene product indicates that YVC1 is primarily localized in the vacuole and not other intracellular membranes. Thus we have identified the YVC1 vacuolar/lysosomal cation-channel gene. This report has implications for the function of TRP channels in other organisms and the possible molecular identification of vacuolar/lysosomal ion channels in other eukaryotes. PMID- 11427714 TI - Intramolecular secondary structure rearrangement by the kissing interaction of the Neurospora VS ribozyme. AB - Kissing interactions in RNA are formed when bases between two hairpin loops pair. Intra- and intermolecular kissing interactions are important in forming the tertiary or quaternary structure of many RNAs. Self-cleavage of the wild-type Varkud satellite (VS) ribozyme requires a kissing interaction between the hairpin loops of stem-loops I and V. In addition, self-cleavage requires a rearrangement of several base pairs at the base of stem I. We show that the kissing interaction is necessary for the secondary structure rearrangement of wild-type stem-loop I. Surprisingly, isolated stem-loop V in the absence of the rest of the ribozyme is sufficient to rearrange the secondary structure of isolated stem-loop I. In contrast to kissing interactions in other RNAs that are either confined to the loops or culminate in an extended intermolecular duplex, the VS kissing interaction causes changes in intramolecular base pairs within the target stem loop. PMID- 11427715 TI - Binding and disruption of phospholipid bilayers by supramolecular RNA complexes. AB - In an RNA world, RNAs would have regulated traffic through normally impermeable bilayer membranes. Using selection-amplification we previously found RNAs that bind stably and increase the ionic conductance of phospholipid membranes at high Mg(2+) and Ca(2+) concentrations. Now selection in reduced divalents yields RNAs that bind phosphatidylcholine liposomes under conditions closer to physiological. Such affinity for phospholipid membranes requires interactions between RNAs. In fact, we detected no functional monomeric membrane-binding RNAs. A membrane active end-to-end heterotrimer consisting of 2 RNA 9 and 1 RNA 10 is defined by nucleotide protection, oligonucleotide competition, and mutant analysis. Oligomers of the heterotrimer bind stably, cause release of liposome-encapsulated solutes, and disrupt model black membranes. Individual RNA molecules do not show any of these activities. This novel mechanism of RNA binding to lipid membranes may not only regulate membrane permeability, but suggests that arrays of catalytic or structural RNAs on membranes are plausible. Finally, a selection met only by RNA complexes evokes new possibilities for selection-amplification itself. PMID- 11427716 TI - Neurotensin-deficient mice show altered responses to antipsychotic drugs. AB - The peptide transmitter neurotensin (NT) exerts diverse neurochemical effects that resemble those seen after acute administration of antipsychotic drugs (APDs). These drugs also induce NT expression in the striatum; this and other convergent findings have led to the suggestion that NT may mediate some APD effects. Here, we demonstrate that the ability of the typical APD haloperidol to induce Fos expression in the dorsolateral striatum is markedly attenuated in NT null mutant mice. The induction of Fos and NT in the dorsolateral striatum in response to typical, but not atypical, APDs has led to the hypothesis that the increased expression of these proteins is mechanistically related to the production of extrapyramidal side effects (EPS). However, we found that catalepsy, which is thought to reflect the EPS of typical APDs, is unaffected in NT-null mutant mice, suggesting that NT does not contribute to the generation of EPS. We conclude that NT is required for haloperidol-elicited activation of a specific population of striatal neurons but not haloperidol-induced catalepsy. These results are consistent with the hypothesis that endogenous NT mediates a specific subset of APD actions. PMID- 11427717 TI - Simple mechanochemistry describes the dynamics of kinesin molecules. AB - Recently, Block and coworkers [Visscher, K., Schnitzer, M. J., & Block, S. M. (1999) Nature (London) 400, 184--189 and Schnitzer, M. J., Visscher, K. & Block, S. M. (2000) Nat. Cell Biol. 2, 718--723] have reported extensive observations of individual kinesin molecules moving along microtubules in vitro under controlled loads, F = 1 to 8 pN, with [ATP] = 1 microM to 2 mM. Their measurements of velocity, V, randomness, r, stalling force, and mean run length, L, reveal a need for improved theoretical understanding. We show, presenting explicit formulae that provide a quantitative basis for comparing distinct molecular motors, that their data are satisfactorily described by simple, discrete-state, sequential stochastic models. The simplest (N = 2)-state model with fixed load-distribution factors and kinetic rate constants concordant with stopped-flow experiments, accounts for the global (V, F, L, [ATP]) interdependence and, further, matches relative acceleration observed under assisting loads. The randomness, r(F,[ATP]), is accounted for by a waiting-time distribution, psi(1)(+)(t), [for the transition(s) following ATP binding] with a width parameter nu identical with (2)/<(Delta t)(2)> approximately 2.5, indicative of a dispersive stroke of mechanicity approximately 0.6 or of a few ( greater than or similar to nu - 1) further, kinetically coupled states: indeed, N = 4 (but not N = 3) models do well. The analysis reveals: (i) a substep of d(0) = 1.8--2.1 nm on ATP binding (consistent with structurally based suggestions); (ii) comparable load dependence for ATP binding and unbinding; (iii) a strong load dependence for reverse hydrolysis and subsequent reverse rates; and (iv) a large ( greater than or similar to 50-fold) increase in detachment rate, with a marked load dependence, following ATP binding. PMID- 11427718 TI - Light-induced phase-delay of the chicken pineal circadian clock is associated with the induction of cE4bp4, a potential transcriptional repressor of cPer2 gene. AB - The chicken pineal gland contains the autonomous circadian oscillator together with the photic-input pathway. We searched for chicken pineal genes that are induced by light in a time-of-day-dependent manner, and isolated chicken homolog of bZIP transcription factor E4bp4 (cE4bp4) showing high similarity to vrille, one of the Drosophila clock genes. cE4bp4 was expressed rhythmically in the pineal gland with a peak at very early (subjective) night under both 12-h light/12-h dark cycle and constant dark conditions, and the phase was nearly opposite to the expression rhythm of cPer2, a chicken pineal clock gene. Luciferase reporter gene assays showed that cE4BP4 represses cPer2 promoter through a E4BP4-recognition sequence present in the 5'-flanking region, indicating that cE4BP4 can down-regulate the chick pineal cPer2 expression. In vivo light-perturbation studies showed that the prolongation of the light period to early subjective night maintained the high level expression of the pineal cE4bp4, and presumably as a consequence delayed the onset of the induction of the pineal cPer2 expression in the next morning. These light-dependent changes in the mRNA levels of the pineal cE4bp4 and cPer2 were followed by a phase-delay of the subsequent cycles of cE4bp4/cPer2 expression, suggesting that cE4BP4 plays an important role in the phase-delaying process as a light-dependent suppressor of cPer2 gene. PMID- 11427719 TI - A cytomegalovirus-encoded inhibitor of apoptosis that suppresses caspase-8 activation. AB - We have identified a human cytomegalovirus cell-death suppressor, denoted vICA, encoded by the viral UL36 gene. vICA inhibits Fas-mediated apoptosis by binding to the pro-domain of caspase-8 and preventing its activation. vICA does not share significant sequence homology with FLIPs or other known suppressors of apoptosis, suggesting that this protein represents a new class of cell-death suppressors. Notably, resistance to Fas-mediated apoptosis is delayed in fibroblasts infected with viruses that encode mutant vICA, suggesting that vICA suppresses death receptor-induced cell death in the context of viral infection. Although vICA is dispensable for viral replication in vitro, the common targeting of caspase-8 activation by diverse herpesviruses argues for an important role for this antiapoptotic mechanism in the pathogenesis of viral infection in the host, most likely in avoiding immune clearance by cytotoxic lymphocytes and natural killer cells. PMID- 11427720 TI - Genetic fidelity under harsh conditions: analysis of spontaneous mutation in the thermoacidophilic archaeon Sulfolobus acidocaldarius. AB - Microbes whose genomes are encoded by DNA and for which adequate information is available display similar genomic mutation rates (average 0.0034 mutations per chromosome replication, range 0.0025 to 0.0046). However, this value currently is based on only a few well characterized microbes reproducing within a narrow range of environmental conditions. In particular, no genomic mutation rate has been determined either for a microbe whose natural growth conditions may extensively damage DNA or for any member of the archaea, a prokaryotic lineage deeply diverged from both bacteria and eukaryotes. Both of these conditions are met by the extreme thermoacidophile Sulfolobus acidocaldarius. We determined the genomic mutation rate for this species when growing at pH 3.5 and 75 degrees C based on the rate of forward mutation at the pyrE gene and the nucleotide changes identified in 101 independent mutants. The observed value of about 0.0018 extends the range of DNA-based microbes with rates close to the standard rate simultaneously to an archaeon and to an extremophile whose cytoplasmic pH and normal growth temperature greatly accelerate the spontaneous decomposition of DNA. The mutations include base pair substitutions (BPSs) and additions and deletions of various sizes, but the S. acidocaldarius spectrum differs from those of other DNA-based organisms in being relatively poor in BPSs. The paucity of BPSs cannot yet be explained by known properties of DNA replication or repair enzymes of Sulfolobus spp. It suggests, however, that molecular evolution per genome replication may proceed more slowly in S. acidocaldarius than in other DNA based organisms examined to date. PMID- 11427721 TI - Role of a pineal cAMP-operated arylalkylamine N-acetyltransferase/14-3-3-binding switch in melatonin synthesis. AB - The daily rhythm in melatonin levels is controlled by cAMP through actions on the penultimate enzyme in melatonin synthesis, arylalkylamine N-acetyltransferase (AANAT; serotonin N-acetyltransferase, EC ). Results presented here describe a regulatory/binding sequence in AANAT that encodes a cAMP-operated binding switch through which cAMP-regulated protein kinase-catalyzed phosphorylation [RRHTLPAN - > RRHpTLPAN] promotes formation of a complex with 14-3-3 proteins. Formation of this AANAT/14-3-3 complex enhances melatonin production by shielding AANAT from dephosphorylation and/or proteolysis and by decreasing the K(m) for 5 hydroxytryptamine (serotonin). Similar switches could play a role in cAMP signal transduction in other biological systems. PMID- 11427722 TI - Deficiency of rds/peripherin causes photoreceptor death in mouse models of digenic and dominant retinitis pigmentosa. AB - Retinitis pigmentosa (RP) is a group of inherited blinding diseases caused by mutations in multiple genes including RDS. RDS encodes rds/peripherin (rds), a 36 kDa glycoprotein in the rims of rod and cone outer-segment (OS) discs. Rom1 is related to rds with similar membrane topology and the identical distribution in OS. In contrast to RDS, no mutations in ROM1 alone have been associated with retinal disease. However, an unusual digenic form of RP has been described. Affected individuals in several families were doubly heterozygous for a mutation in RDS causing a leucine 185 to proline substitution in rds (L185P) and a null mutation in ROM1. Neither mutation alone caused clinical abnormalities. Here, we generated transgenic/knockout mice that duplicate the amino acid substitutions and predicted levels of rds and rom1 in patients with RDS-mediated digenic and dominant RP. Photoreceptor degeneration in the mouse model of digenic RP was faster than in the wild-type and monogenic controls by histological, electroretinographic, and biochemical analysis. We observed a positive correlation between the rate of photoreceptor loss and the extent of OS disorganization in mice of several genotypes. Photoreceptor degeneration in RDS mediated RP appears to be caused by a simple deficiency of rds and rom1. The critical threshold for the combined abundance of rds and rom1 is approximately 60% of wild type. Below this value, the extent of OS disorganization results in clinically significant photoreceptor degeneration. PMID- 11427723 TI - Catalytic consumption of nitric oxide by 12/15- lipoxygenase: inhibition of monocyte soluble guanylate cyclase activation. AB - 12/15-Lipoxygenase (LOX) activity is elevated in vascular diseases associated with impaired nitric oxide (( small middle dot)NO) bioactivity, such as hypertension and atherosclerosis. In this study, primary porcine monocytes expressing 12/15-LOX, rat A10 smooth muscle cells transfected with murine 12/15 LOX, and purified porcine 12/15-LOX all consumed *NO in the presence of lipid substrate. Suppression of LOX diene conjugation by *NO was also found, although the lipid product profile was unchanged. *NO consumption by porcine monocytes was inhibited by the LOX inhibitor, eicosatetraynoic acid. Rates of arachidonate (AA) or linoleate (LA)-dependent *NO depletion by porcine monocytes (2.68 +/- 0.03 nmol x min(-1) x 10(6) cells(-1) and 1.5 +/- 0.25 nmol x min(-1) x 10(6) cells( 1), respectively) were several-fold greater than rates of *NO generation by cytokine-activated macrophages (0.1-0.2 nmol x min(-1) x 10(6) cells(-1)) and LA dependent *NO consumption by primary porcine monocytes inhibited *NO activation of soluble guanylate cyclase. These data indicate that catalytic *NO consumption by 12/15-LOX modulates monocyte *NO signaling and suggest that LOXs may contribute to vascular dysfunction not only by the bioactivity of their lipid products, but also by serving as catalytic sinks for *NO in the vasculature. PMID- 11427724 TI - Different modes of hippocampal plasticity in response to estrogen in young and aged female rats. AB - Estrogen regulates hippocampal dendritic spine density and synapse number in an N methyl-D-aspartate (NMDA) receptor-dependent manner, and these effects may be of particular importance in the context of age-related changes in endocrine status. We investigated estrogen's effects on axospinous synapse density and the synaptic distribution of the NMDA receptor subunit, NR1, within the context of aging. Although estrogen induced an increase in axospinous synapse density in young animals, it did not alter the synaptic representation of NR1, in that the amount of NR1 per synapse was equivalent across groups. Estrogen replacement in aged female rats failed to increase axospinous synapse density; however, estrogen up regulated synaptic NR1 compared with aged animals with no estrogen. Therefore, the young and aged hippocampi react differently to estrogen replacement, with the aged animals unable to mount a plasticity response generating additional synapses, yet responsive to estrogen with respect to additional NMDA receptor content per synapse. These findings have important implications for estrogen replacement therapy in the context of aging. PMID- 11427725 TI - Identification of a subpopulation of rapidly self-renewing and multipotential adult stem cells in colonies of human marrow stromal cells. AB - Marrow stromal cells are adult stem cells from bone marrow that can differentiate into multiple nonhematopoietic cell lineages. Previous reports demonstrated that single-cell-derived colonies of marrow stromal cells contained two morphologically distinct cell types: spindle-shaped cells and large flat cells. Here we found that early colonies also contain a third kind of cell: very small round cells that rapidly self-renew. Samples enriched for the small cells had a greater potential for multipotential differentiation than samples enriched for the large cells. Also, the small cells expressed a series of surface epitopes and other proteins that potentially can be used to distinguish the small cells from the large cells. The results suggested it will be important to distinguish the major subpopulations of marrow stromal cells in defining their biology and their potential for cell and gene therapy. PMID- 11427726 TI - The complete genome of the crenarchaeon Sulfolobus solfataricus P2. AB - The genome of the crenarchaeon Sulfolobus solfataricus P2 contains 2,992,245 bp on a single chromosome and encodes 2,977 proteins and many RNAs. One-third of the encoded proteins have no detectable homologs in other sequenced genomes. Moreover, 40% appear to be archaeal-specific, and only 12% and 2.3% are shared exclusively with bacteria and eukarya, respectively. The genome shows a high level of plasticity with 200 diverse insertion sequence elements, many putative nonautonomous mobile elements, and evidence of integrase-mediated insertion events. There are also long clusters of regularly spaced tandem repeats. Different transfer systems are used for the uptake of inorganic and organic solutes, and a wealth of intracellular and extracellular proteases, sugar, and sulfur metabolizing enzymes are encoded, as well as enzymes of the central metabolic pathways and motility proteins. The major metabolic electron carrier is not NADH as in bacteria and eukarya but probably ferredoxin. The essential components required for DNA replication, DNA repair and recombination, the cell cycle, transcriptional initiation and translation, but not DNA folding, show a strong eukaryal character with many archaeal-specific features. The results illustrate major differences between crenarchaea and euryarchaea, especially for their DNA replication mechanism and cell cycle processes and their translational apparatus. PMID- 11427727 TI - Expression of error-prone polymerases in BL2 cells activated for Ig somatic hypermutation. AB - High affinity antibodies are generated in mice and humans by means of somatic hypermutation (SHM) of variable (V) regions of Ig genes. Mutations with rates of 10(-5)--10(-3) per base pair per generation, about 10(6)-fold above normal, are targeted primarily at V-region hot spots by unknown mechanisms. We have measured mRNA expression of DNA polymerases iota, eta, and zeta by using cultured Burkitt's lymphoma (BL)2 cells. These cells exhibit 5-10-fold increases in heavy chain V-region mutations targeted only predominantly to RGYW (R = A or G, Y = C or T, W = T or A) hot spots if costimulated with T cells and IgM crosslinking, the presumed in vivo requirements for SHM. An approximately 4-fold increase pol iota mRNA occurs within 12 h when cocultured with T cells and surface IgM crosslinking. Induction of pols eta and zeta occur with T cells, IgM crosslinking, or both stimuli. The fidelity of pol iota was measured at RGYW hot- and non-hot-spot sequences situated at nicks, gaps, and double-strand breaks. Pol iota formed T x G mispairs at a frequency of 10(-2), consistent with SHM generated C to T transitions, with a 3-fold increased error rate in hot- vs. non hot-spot sequences for the single-nucleotide overhang. The T cell and IgM crosslinking-dependent induction of pol iota at 12 h may indicate an SHM "triggering" event has occurred. However, pols iota, eta, and zeta are present under all conditions, suggesting that their presence is not sufficient to generate mutations because both T cell and IgM stimuli are required for SHM induction. PMID- 11427728 TI - Raf-1 promotes cell survival by antagonizing apoptosis signal-regulating kinase 1 through a MEK-ERK independent mechanism. AB - The Ser/Thr kinase Raf-1 is a protooncogene product that is a central component in many signaling pathways involved in normal cell growth and oncogenic transformation. Upon activation, Raf-1 phosphorylates mitogen-activated protein kinase kinase (MEK), which in turn activates mitogen-activated protein kinase/extracellular signal-regulated kinases (MAPK/ERKs), leading to the propagation of signals. Depending on specific stimuli and cellular environment, the Raf-1--MEK--ERK cascade regulates diverse cellular processes such as proliferation, differentiation, and apoptosis. Here, we describe a MEK--ERK independent prosurvival function of Raf-1. We found that Raf-1 interacts with the proapoptotic, stress-activated protein kinase ASK1 (apoptosis signal-regulating kinase 1) in vitro and in vivo. Deletion analysis localized the Raf-1 binding site to the N-terminal regulatory fragment of ASK1. This interaction allows Raf-1 to act independently of the MEK--ERK pathway to inhibit apoptosis. Furthermore, catalytically inactive forms of Raf-1 can mimic the wild-type effect, raising the possibility of a kinase-independent function of Raf-1. Thus, Raf-1 may promote cell survival through its protein-protein interactions in addition to its established MEK kinase function. PMID- 11427729 TI - Onset of natural killer cell lymphomas in transgenic mice carrying a truncated HMGI-C gene by the chronic stimulation of the IL-2 and IL-15 pathway. AB - Rearrangements of the high mobility group protein I-C (HMGI-C) gene, consisting in the loss of the carboxyl-terminal tail, have been frequently detected in benign human tumors of mesenchymal origin. We have previously demonstrated that transgenic (TG) mice carrying a truncated HMGI-C construct (HMGI-C/T) exhibit a giant phenotype together with a predominantly abdominal/pelvic lipomatosis. Here, we report that HMGI-C/T TG mice develop natural killer (NK)-T/NK cell lymphomas starting from 12 months of age. We found an increased expression of IL-2 and IL 15 proteins and their receptors in these lymphomas, and we demonstrate that HMGI C/T protein positively regulates their expression in vitro. Therefore, the HMGI C/T-mediated chronic stimulation of the IL-2/IL-15 pathway could be responsible for the onset of NK-T/NK cell lymphomas in HMGI-C/T TG mice. PMID- 11427730 TI - In vitro characterization of DNA gyrase inhibition by microcin B17 analogs with altered bisheterocyclic sites. AB - Microcin B17 (MccB17) is a 3.1-kDa Escherichia coli antibiotic that contains thiazole and oxazole heterocycles in a peptide backbone. MccB17 inhibits its cellular target, DNA gyrase, by trapping the enzyme in a complex that is covalently bound to double-strand cleaved DNA, in a manner similar to the well known quinolone drugs. The identification of gyrase as the target of MccB17 provides an opportunity to analyze the relationship between the structure of this unusual antibiotic and its activity. In this report, steady-state parameters are used to describe the induction of the cleavable complex by MccB17 analogs containing modified bisheterocyclic sites. The relative potency of these analogs corresponds to the capacity of the compounds to prevent growth of sensitive cells. In contrast to previously reported experiments, inhibition of DNA gyrase supercoiling activity by wild-type MccB17 also was observed. These results suggest that DNA gyrase is the main intracellular target of MccB17. This study probes the structure-function relationship of a new class of gyrase inhibitors and demonstrates that these techniques could be used to analyze compounds in the search for clinically useful antibiotics that block DNA gyrase. PMID- 11427731 TI - Altered peptide ligand vaccination with Flt3 ligand expanded dendritic cells for tumor immunotherapy. AB - Most tumor-associated antigens represent self-proteins and as a result are poorly immunogenic due to immune tolerance. Here we show that tolerance to carcinoembryonic antigen (CEA), which is overexpressed by the majority of lethal malignancies, can be reversed by immunization with a CEA-derived peptide. This peptide was altered to make it a more potent T cell antigen and loaded onto dendritic cells (DCs) for delivery as a cellular vaccine. Although DCs are rare in the blood, we found that treatment of advanced cancer patients with Flt3 ligand, a hematopoietic growth factor, expanded DCs 20-fold in vivo. Immunization with these antigen-loaded DCs induced CD8 cytotoxic T lymphocytes that recognized tumor cells expressing endogenous CEA. Staining with peptide-MHC tetramers demonstrated the expansion of CD8 T cells that recognize both the native and altered epitopes and possess an effector cytotoxic T lymphocyte phenotype (CD45RA(+)CD27(-)CCR7(-)). After vaccination, two of 12 patients experienced dramatic tumor regression, one patient had a mixed response, and two had stable disease. Clinical response correlated with the expansion of CD8 tetramer(+) T cells, confirming the role of CD8 T cells in this treatment strategy. PMID- 11427732 TI - Xenopus Bcl-X(L) selectively protects Rohon-Beard neurons from metamorphic degeneration. AB - Amphibian metamorphosis involves extensive, but selective, neuronal death and turnover, thus sharing many features with mammalian postnatal development. The antiapoptotic protein Bcl-X(L) plays an important role in postnatal mammalian neuronal survival. It is therefore of interest that accumulation of the mRNA encoding the Xenopus Bcl-X(L) homologue, termed xR11, increases abruptly in the nervous system, but not in other tissues, during metamorphosis in Xenopus tadpoles. This observation raises the intriguing possibility that xR11 selectively regulates neuronal survival during postembryonic development. To investigate this hypothesis, we overexpressed xR11 in vivo as a green fluorescent protein (GFP)-xR11 fusion protein by using somatic and germinal transgenesis. Somatic gene transfer showed that the fusion protein was effective in counteracting, in a dose-dependent manner, the proapoptotic effects of coexpressed Bax. When GFP-xR11 was expressed from the neuronal beta-tubulin promoter by germinal transgenesis we observed neuronal specific expression that was maintained throughout metamorphosis and beyond, into juvenile and adult stages. Confocal microscopy showed GFP-xR11 to be exclusively localized in the mitochondria. Our findings show that GFP-xR11 significantly prolonged Rohon-Beard neuron survival up to the climax of metamorphosis, even in the regressing tadpole tail, whereas in controls these neurons disappeared in early metamorphosis. However, GFP-xR11 expression did not modify the fate of spinal cord motoneurons. The selective protection of Rohon-Beard neurons reveals cell-specific apoptotic pathways and offers approaches to further analyze programmed neuronal turnover during postembryonic development. PMID- 11427733 TI - Reversible encapsulation by self-assembling resorcinarene subunits. AB - Encapsulation complexes are assemblies in which a reversibly formed host more or less completely surrounds guest molecules. Host structures held together by hydrogen bonds have lifetimes in organic solvents of milliseconds to hours, long enough to directly observe the encapsulated guest by NMR spectroscopy. We describe here the action of alkyl ammonium compounds as guests that gather up to six molecules of the host module to form encapsulation complexes. The stoichiometry of the complexes--the largest hydrogen-bonded host capsules to date -is determined by the size and concentration of the guest. PMID- 11427734 TI - The Caenorhabditis elegans hif-1 gene encodes a bHLH-PAS protein that is required for adaptation to hypoxia. AB - Hypoxia-inducible factor, a heterodimeric transcription complex, regulates cellular and systemic responses to low oxygen levels (hypoxia) during normal mammalian development or tumor progression. Here, we present evidence that a similar complex mediates response to hypoxia in Caenorhabditis elegans. This complex consists of HIF-1 and AHA-1, which are encoded by C. elegans homologs of the hypoxia-inducible factor (HIF) alpha and beta subunits, respectively. hif-1 mutants exhibit no severe defects under standard laboratory conditions, but they are unable to adapt to hypoxia. Although wild-type animals can survive and reproduce in 1% oxygen, the majority of hif-1-defective animals die in these conditions. We show that the expression of an HIF-1:green fluorescent protein fusion protein is induced by hypoxia and is subsequently reduced upon reoxygenation. Both hif-1 and aha-1 are expressed in most cell types, and the gene products can be coimmunoprecipitated. We conclude that the mechanisms of hypoxia signaling are likely conserved among metazoans. Additionally, we find that nuclear localization of AHA-1 is disrupted in an hif-1 mutant. This finding suggests that heterodimerization may be a prerequisite for efficient nuclear translocation of AHA-1. PMID- 11427735 TI - Id1 regulation of cellular senescence through transcriptional repression of p16/Ink4a. AB - The Id family of helix-loop-helix (HLH) transcriptional regulatory proteins does not possess a basic DNA-binding domain and functions as a negative regulator of basic HLH transcription factors. Id proteins coordinate cell growth and differentiation pathways within mammalian cells and have been shown to regulate G(1)-S cell-cycle transitions. Although much recent data has implicated Id1 in playing a critical role in modulating cellular senescence, no direct genetic evidence has been reported to substantiate such work. Here we show that Id1-null primary mouse embryo fibroblasts undergo premature senescence despite normal growth profiles at early passage. These cells possess increased expression of the tumor-suppressor protein p16/Ink4a but not p19/ARF, and have decreased cyclin dependent kinase (cdk) 2 and cdk4 kinase activity. We also show that Id1 is able to directly inhibit p16/Ink4a but not p19/ARF promoter activity via its HLH domain, and that Id1 inhibits transcriptional activation at E-boxes within the p16/Ink4a promoter. Our data provide, to our knowledge, the first genetic evidence for a role for Id1 as an inhibitor of cellular senescence and suggest that Id1 functions to delay cellular senescence through repression of p16/Ink4a. Because epigenetic and genetic abrogation of p16/Ink4a function has been implicated in the evolution of several human malignancies, we propose that transcriptional regulation of p16/Ink4a may also provide a mechanism for the dysregulation of normal cellular growth controls during the evolution of human malignancies. PMID- 11427736 TI - Electroretinogram analysis of relative spectral sensitivity in genetically identified dichromatic macaques. AB - The retinas of macaque monkeys usually contain three types of photopigment, providing them with trichromatic color vision homologous to that of humans. However, we recently used molecular genetic analysis to identify several macaques with a dichromatic genotype. The affected X chromosome of these animals contains a hybrid gene of long-wavelength-sensitive (L) and middle-wavelength-sensitive (M) photopigments instead of separate genes encoding L and M photopigments. The product of the hybrid gene exhibits a spectral sensitivity close to that of M photopigment; consequently, male monkeys carrying the hybrid gene are genetic protanopes, effectively lacking L photopigment. In the present study, we assessed retinal expression of L photopigment in monkeys carrying the hybrid gene. The relative sensitivities to middle-wavelength (green) and long-wavelength (red) light were measured by electroretinogram flicker photometry. We found the sensitivity to red light to be extremely low in protanopic male monkeys compared with monkeys with the normal genotype. In female heterozygotes, sensitivity to red light was intermediate between the genetic protanopes and normal monkeys. Decreased sensitivity to long wavelengths was thus consistent with genetic loss of L photopigment. PMID- 11427737 TI - Metabolic engineering of essential oil yield and composition in mint by altering expression of deoxyxylulose phosphate reductoisomerase and menthofuran synthase. AB - Peppermint (Mentha x piperita L.) was independently transformed with a homologous sense version of the 1-deoxy-d-xylulose-5-phosphate reductoisomerase cDNA and with a homologous antisense version of the menthofuran synthase cDNA, both driven by the CaMV 35S promoter. Two groups of transgenic plants were regenerated in the reductoisomerase experiments, one of which remained normal in appearance and development; another was deficient in chlorophyll production and grew slowly. Transgenic plants of normal appearance and growth habit expressed the reductoisomerase transgene strongly and constitutively, as determined by RNA blot analysis and direct enzyme assay, and these plants accumulated substantially more essential oil (about 50% yield increase) without change in monoterpene composition compared with wild-type. Chlorophyll-deficient plants did not afford detectable reductoisomerase mRNA or enzyme activity and yielded less essential oil than did wild-type plants, indicating cosuppression of the reductoisomerase gene. Plants transformed with the antisense version of the menthofuran synthase cDNA were normal in appearance but produced less than half of this undesirable monoterpene oil component than did wild-type mint grown under unstressed or stressed conditions. These experiments demonstrate that essential oil quantity and quality can be regulated by metabolic engineering. Thus, alteration of the committed step of the mevalonate-independent pathway for supply of terpenoid precursors improves flux through the pathway that leads to increased monoterpene production, and antisense manipulation of a selected downstream monoterpene biosynthetic step leads to improved oil composition. PMID- 11427738 TI - Aromatase (Cyp19) expression is up-regulated by targeted disruption of Dax1. AB - DAX-1 [dosage-sensitive sex reversal, adrenal hypoplasia congenita (AHC) critical region on the X chromosome, gene 1] is an orphan nuclear receptor that represses transcription by steroidogenic factor-1 (SF-1), a factor that regulates expression of multiple steroidogenic enzymes and other genes involved in reproduction. Mutations in the human DAX1 gene (also known as AHC) cause the X linked syndrome AHC, a disorder that is associated with hypogonadotropic hypogonadism also. Characterization of Dax1-deficient male mice revealed primary testicular defects that included Leydig cell hyperplasia (LCH) and progressive degeneration of the germinal epithelium, leading to infertility. In this study, we investigated the effect of Dax1 disruption on the expression profile of various steroidogenic enzyme genes in Leydig cells isolated from Dax1-deficient male mice. Expression of the aromatase (Cyp19) gene, which encodes the enzyme that converts testosterone to estradiol, was increased significantly in the Leydig cells isolated from mutant mice, whereas the expression of other proteins (e.g., StAR and Cyp11a) was not altered. In in vitro transfection studies, DAX-1 repressed the SF-1-mediated transactivation of the Cyp19 promoter but did not inhibit the StAR or Cyp11a promoters. Elevated Cyp19 expression was accompanied by increased intratesticular levels of estradiol. Administration of tamoxifen, a selective estrogen-receptor modulator, restored fertility to the Dax1-deficient male mice and partially corrected LCH, suggesting that estrogen excess contributes to LCH and infertility. Based on these in vivo and in vitro analyses, aromatase seems to be a physiologic target of Dax-1 in Leydig cells, and increased Cyp19 expression may account, in part, for the infertility and LCH in Dax1-deficient mice. PMID- 11427739 TI - Induction of primordial germ cells from murine epiblasts by synergistic action of BMP4 and BMP8B signaling pathways. AB - Extraembryonic ectoderm-derived factors instruct the pluripotent epiblast cells to develop toward a restricted primordial germ cell (PGC) fate during murine gastrulation. Genes encoding Bmp4 of the Dpp class and Bmp8b of the 60A class are expressed in the extraembryonic ectoderm and targeted mutation of either results in severe defects in PGC formation. It has been shown that heterodimers of DPP and 60A classes of bone morphogenetic proteins (BMPs) are more potent than each homodimers in bone and mesoderm induction in vitro, suggesting that BMP4 and BMP8B may form heterodimers to induce PGCs. To investigate how BMP4 and BMP8B interact and signal for PGC induction, we cocultured epiblasts of embryonic day 6.0--6.25 embryos with BMP4 and BMP8B proteins produced by COS cells. Our data show that BMP4 or BMP8B homodimers alone cannot induce PGCs whereas they can in combination, providing evidence that two BMP pathways are simultaneously required for the generation of a given cell type in mammals and also providing a prototype method for PGC induction in vitro. Furthermore, the PGC defects of Bmp8b mutants can be rescued by BMP8B homodimers whereas BMP4 homodimers cannot mitigate the PGC defects of Bmp4 null mutants, suggesting that BMP4 proteins are also required for epiblast cells to gain germ-line competency before the synergistic action of BMP4 and BMP8B. PMID- 11427741 TI - Human herpesviruses 6 and 7: effects on hematopoiesis and mode of transmission. AB - Human herpesvirus 6 (HHV-6) and human herpesvirus 7 (HHV-7) were recently discovered, and are known as etiologic agents of exanthem subitum (roseola). HHV 6 and HHV-7 are T-lymphotropic, and have been classified as betaherpesviruses. In monitoring of herpesviruses after hematopoietic stem cell transplantation, each herpesvirus had a unique temporal profile of detection. HHV-6 DNA was detected most frequently at 3 weeks, whereas cytomegalovirus and Epstein-Barr virus DNA were detected later. HHV-7 DNA was not detected throughout the observation period. In in vitro hematopoietic colony assays, HHV-6 suppressed all three lineages of hematopoiesis, i.e., erythroid, granulocyte/macrophage, and megakaryocyte, whereas HHV-7 did not have any suppressive effect. Molecular epidemiological analysis revealed that HHV-7 was transmitted horizontally from grandparents to parents to children through close contact within a household. Either parent could transmit HHV-7 to the children. Follow-up studies of the amount of viral DNA in saliva samples revealed that the amount of HHV-7 DNA was rather constant for each individual, and that "high producers" and "low producers" could be distinguished. Transferred antibodies against HHV-7 tended to be higher and remain longer after birth than those of HHV-6, and these findings are consistent with the clinical observation that HHV-6 infection occurs earlier than HHV-7 infection. PMID- 11427742 TI - GB virus-C/hepatitis G virus. AB - GB virus-C (GBV-C)/hepatitis G virus (HGV) is a positive, single-strand RNA virus that has been classified in the family Flaviviridae. Interestingly, GBV-C/HGV appears to have a truncated or absent core protein at the amino terminus of the polyprotein. GBV-C/HGV is transmitted parenterally and probably sexually. Most GBV-C/HGV infections appear to be asymptomatic, persistent, and no correlation between virus infection and liver dysfunction although the disease-inducing activity of GBV-C/HGV remains to be investigated. Furthermore, there was no evidence of pathogenesis in the liver by experiment with chimpanzees. From these results, GBV-C/HGV might be considered as a kind of "orphan" virus in search of a disease. Epidemiological investigation demonstrated that GBV-C/HGV infection is present in about 1-1.4% of the healthy population in developed countries and in 8 14.6% in developing countries. The genome of GBV-C/HGV exhibits a sequence variation among different isolates. On the basis of this variation, it has been proposed that GBV-C/HGV can be classified into at least four major genotypes, consisting of type 1 (West Africa), type 2 (US/Europe), type 3 (Asia), and type 4 (Southeast Asia). PMID- 11427743 TI - Prevalence of resistance to antimicrobials of Escherichia coli isolates from clinical sources at a private hospital in Trinidad. AB - Antimicrobial susceptibility patterns of strains of Escherichia coli isolated between 1994 and 1998 were studied. Of the 1,283 strains examined, 75% were recovered from urine, 8.7% from wounds, 3.2% from blood, 2.6% from pus, and 10.5% from other sources. Isolates from inpatients and outpatients accounted for 46.1% and 53.9%, respectively. Gentamicin and nalidixic acid showed the greatest efficacy against isolates from both inpatients and outpatients, revealing a >90% sensitivity. Drugs with the lowest efficacies were ampicillin and amoxicillin clavulanic acid, which showed a >45% resistance. Tetracycline showed a significant decline in resistance from 1994 to 1998 among strains from both inpatients and outpatients (P < 0.001). This decline may be related to a policy of restrictive antibiotic reporting by the Microbiology Laboratory and seminars for general practitioners, subsequent to an island-wide survey an antibiotic resistance. A similar pattern of declining resistance was also observed for cefuroxime. E. coli sensitivity to co-trimoxazole was relatively stable during the study period. Although the overall prevalence of resistance among E. coli strains is relatively low, on-going surveillance of bacterial resistance must continue. The microbial antibiogram can provide general practitioners and clinicians with data essential for optimum empiric choices. Further, the introduction of a policy of restrictive reporting may act "synergistically" with the education of doctors on resistance patterns, to effect island-wide reduction of antimicrobial resistance. PMID- 11427744 TI - Periodontitis and serum interleukin-6 levels in the elderly. AB - The elderly lose teeth as a result of dental caries and periodontitis caused by pathogenic oral bacteria. Periodontitis produces inflammatory cytokines due to the presence of lipopolysaccharides from oral gram-negative bacteria. Although the number of circulating inflammatory cytokines is related to the severity of the periodontitis, it is unclear whether the concentrations also correlate with periodontitis in the elderly. We investigated the relationship between periodontitis status and the concentrations of serum interleukin-6 (IL-6) in the serum from 276 subjects of 70- and 80-year-olds. Of the 276 subjects, 227 (82%) were dentate, 149 (54%) were found to be positive for serum IL-6, and 29 (13%) of the dentate subjects had severe periodontitis. However, there were no significant differences between the severity of periodontitis or the number of teeth and the mean serum IL-6 concentrations. These results provided no evidence to support an association between circulating IL-6 and periodontitis in the elderly. PMID- 11427745 TI - Detection of Giardia lamblia cysts in non-fixed long-term stored human feces by direct immunofluorescence assay. AB - Giardia lamblia cysts in fecal specimens are detected by conventional morphological methods. The direct immunofluorescence monoclonal antibody assay (DFA) is also applied to detect G. lamblia cysts in feces, but little is known about the usefulness of DFA in fecal specimens stored under various conditions. The aim of the present study was to evaluate the DFA for detection of G. lamblia cysts and to compare these results with the direct smear method in long-term storage of non-fixed fecal specimens. Fecal specimens with G. lamblia cysts were stored in a refrigerator at 5 degrees C(14 samples), a freezer at -20 degrees C (9 samples), or in 3.9% formalin-saline solution at room temperature (28 samples). G. lamblia cysts were detected by DFA in all stored specimens, while they were detected in only 56% of refrigerated and 93% of frozen specimens by the direct smear method. The storage period of all samples testing negative by direct smear was more than 24 months. Many degenerated cysts were recognized by DFA when cysts were negative by the direct smear method. Our results indicate that DFA is a useful method for detecting G. lamblia cysts in fecal samples that have undergone long-term storage. PMID- 11427746 TI - Detection of anti-Coxiella burnetii antibodies by immunofluorescent technique and enzyme-linked immunosorbent assay. PMID- 11427747 TI - Epidemiological survey of beta-hemolytic streptococci isolated from acute pharyngitis in a private pediatric practice. PMID- 11427748 TI - Autism and infection/immunization episodes in Japan. PMID- 11427749 TI - Tertian malaria outbreak three decades after its eradication. PMID- 11427750 TI - Surveillance of poliovirus-isolates in Japan, 2000. PMID- 11427751 TI - Molecular epidemiology of methicillin-resistant Staphylococcus aureus in a hospital in Chiang Mai. PMID- 11427752 TI - Further simulation on survival of mutants under non-selective condition. PMID- 11427753 TI - Seroepidemiological study of human Parechovirus 1. PMID- 11427754 TI - Air quality monitoring in a neonatal intensive care unit. PMID- 11427755 TI - Outbreak of enterohemorrhagic Escherichia coli O157 mass infection caused by "whole roasted cow". PMID- 11427756 TI - GB virus C/hepatitis G virus does not induce expression of p44 antigen in chimpanzee hepatocytes. PMID- 11427757 TI - Molecular epidemiology of methicillin-resistant Staphylococcus aureus in a Tokyo hospital in 2000. PMID- 11427758 TI - Physical activity and health: introduction to the dose-response symposium. PMID- 11427759 TI - Dose-response issues concerning physical activity and health: an evidence-based symposium. PMID- 11427760 TI - Analyzing the relationship of exercise and health: methods, assumptions, and limitations. AB - The papers in this supplement to Medicine and Science in Sports and Exercise seek to summarize our knowledge of the dose-response relationship of exercise with a variety of health outcomes. This type of rigorous evidence summation has become quite popular in the past 10 years as health care providers and policymakers seek to distinguish interventions that are helpful--the expected benefits justify human and economic costs--from those of no or uncertain benefit (7). Proponents of "evidence-based medicine" argue that in contrast to old-style literature reviews in which the writer, typically an "expert," develops a thesis and cites only those publications that support his point of view, an evidence-based approach finds all of the evidence and objectively weighs it to determine the "truth" (20,22). Although such methods offer the promise of increased objectivity, they are not immune to bias, and readers should understand that the declaration that something is "evidence-based" is no guarantee of its veracity. In fact, because the literature virtually never includes studies that perfectly match the setting and personal circumstance of the patient under treatment, "evidence-informed medicine" would be a more honest title for care grounded in systematic reviews of the literature (23). This paper begins with a discussion of potential theoretical problems with systematic reviews and concludes by considering specific problems that members of the panel were likely to encounter when evaluating the literature on exercise and health. These concepts were presented in the initial lecture of the conference so that the 24 lecturers and 6 members of the consensus panel would be forewarned of the dangers that lay ahead. PMID- 11427761 TI - Type of activity: resistance, aerobic and leisure versus occupational physical activity. AB - PURPOSE: To define and describe the essential terminology associated with dose response issues in physical activity and health. METHODS: Recent consensus documents, position stands, and reports were used to provide reference definitions and methods of classifying physical activity and exercise. RESULTS: The two principal categories of physical activity are occupational physical activity (OPA) and leisure-time physical activity (LTPA). OPA is usually referenced to an 8-h d, whereas the duration of LTPA is quite variable. LTPA includes all forms of aerobic activities, structured endurance exercise programs, resistance-training programs, and sports. Energy expenditure associated with aerobic activity can be expressed in absolute terms (kJ x min(-1)), referenced to body mass (METs), or relative to some maximal physiological response (i.e., maximal heart rate (HR) or aerobic power (VO(2max))). The net cost of physical activity should be used to express energy expenditure relative to dose-response issues. The intensity of resistance training is presented in terms relative to the greatest weight that can be lifted one time in good form (1RM). The intensity of OPA followed the guidance of a previous consensus conference. The intensity of most LTPA can be categorized using the standard aerobic exercise classifications; however, for long-duration (2+ hours) LTPA, the classifications for OPA may be more appropriate. CONCLUSION: Physical activities should be classified in a consistent and standardized manner in terms of both energy expenditure and the relative effort required. PMID- 11427762 TI - Quantifying energy expenditure and physical activity in the context of dose response. AB - PURPOSE: Methods for assessing physical activity (PA) and energy expenditure (EE) were reviewed to identify potential limitations to evaluating and interpreting dose-response relationships between PA and health-related outcomes and to suggest future research directions in this area. METHODS: Literature describing PA and EE assessment methodology was reviewed according to the reported validity, reliability, and feasibility of the measurement in epidemiologic studies. A summary of this review is presented for techniques applicable to studying PA or EE among free-living individuals. RESULTS: Several methods with varying degrees of precision and feasibility have been used to assess PA and EE in free-living populations. Lack of a gold standard field measure of PA may explain some of the variability in precision among these methods. The most accurate field measure of EE appears to be doubly labeled water; however, this approach has limited feasibility in terms of cost and use in studies of total EE only. Electronic motion sensors and physiologic measures related with EE are limited in their ability to discriminate specific types of PA and by inconvenient measurement procedures. Self-reported PA records and surveys are low-cost, relatively unobtrusive methods of assessing PA and EE in field settings and vary in terms of their format, mode of administration, and degree of detailing habitual PA levels. Disparity in the metric used to quantify PA and EE exists within the current literature, which limits the interpretation and comparison of observed dose response relationships. CONCLUSIONS: Efforts to develop equated methods of assessing PA and EE in free-living populations are needed before a systematic evaluation and interpretation of dose-response characteristics between PA and specific health-related parameters can be undertaken. PMID- 11427763 TI - Is physical activity or physical fitness more important in defining health benefits? AB - PURPOSE: We addressed three questions: 1) Is there a dose-response relation between physical activity and health? 2) Is there a dose-response relation between cardiorespiratory fitness and health? 3) If both activity and fitness have a dose-response relation to health, is it possible to determine which exposure is more important? METHODS: We identified articles by PubMed search (restricted from 1/1/90 to 8/25/00) using keywords related to physical activity, physical fitness, and health. An author scanned titles and abstracts of 9831 identified articles. We included for thorough review articles that included three or more categories of activity or fitness and a health outcome and excluded articles on clinical trials, review papers, comments, letters, case reports, and nonhuman studies. We used an evidence-based approach to evaluate the quality of the published data. RESULTS: We summarized results from 67 articles meeting final selection criteria. There is good consensus across studies with most showing an inverse dose-response gradient across both activity and fitness categories for morbidity from coronary heart disease (CHD), stroke, cardiovascular disease (CVD), or cancer; and for CVD, cancer, or all-cause mortality. CONCLUSIONS: All studies reviewed were prospective observational investigations; thus, conclusions are based on Evidence Category C. 1) There is a consistent gradient across activity groups indicating greater longevity and reduced risk of CHD, CVD, stroke, and colon cancer in more active individuals. 2) Studies are compelling in the consistency and steepness of the gradient across fitness groups. Most show a curvilinear gradient, with a steep slope at low levels of fitness and an asymptote in the upper part of the fitness distribution. 3) It is not possible to conclude whether activity or fitness is more important for health. Future studies should define more precisely the shape of the dose-response gradient across activity or fitness groups, evaluate the role of musculoskeletal fitness, and investigate additional health outcomes. PMID- 11427764 TI - Absolute versus relative intensity of physical activity in a dose-response context. AB - PURPOSE: To examine the importance of relative versus absolute intensities of physical activity in the context of population health. METHODS: A standard computer-search of the literature was supplemented by review of extensive personal files. RESULTS: Consensus reports (Category D Evidence) have commonly recommended moderate rather than hard physical activity in the context of population health. Much of the available literature provides Category C Evidence. It has often confounded issues of relative intensity with absolute intensity or total weekly dose of exercise. In terms of cardiovascular health, there is some evidence for a threshold intensity of effort, perhaps as high as 6 METs, in addition to a minimum volume of physical activity. Decreases in blood pressure and prevention of stroke seem best achieved by moderate rather than high relative intensities of physical activity. Many aspects of metabolic health depend on the total volume of activity; moderate relative intensities of effort are more effective in mobilizing body fat, but harder relative intensities may help to increase energy expenditures postexercise. Hard relative intensities seem needed to augment bone density, but this may reflect an associated increase in volume of activity. Hard relative intensities of exercise induce a transient immunosuppression. The optimal intensity of effort, relative or absolute, for protection against various types of cancer remains unresolved. Acute effects of exercise on mood state also require further study; long-term benefits seem associated with a moderate rather than a hard relative intensity of effort. CONCLUSIONS: The importance of relative versus absolute intensity of effort depends on the desired health outcome, and many issues remain to be resolved. Progress will depend on more precise epidemiological methods of assessing energy expenditures and studies that equate total energy expenditures between differing relative intensities. There is a need to focus on gains in quality-adjusted life expectancy. PMID- 11427766 TI - Issues of fractionization of exercise (short vs long bouts). AB - PURPOSE: To evaluate evidence comparing the influence on health outcomes of different patterns and intensities of exercise with equivalent total energy expenditure. METHODS: A computerized literature search, with searches of the reference lists of papers identified. RESULTS: Studies fell into two categories: 1) comparisons of one continuous session of exercise with several short (> or = 10-min) sessions of the same total duration; and 2) comparisons of a session of moderate/hard exercise with a session of lower intensity but equivalent energy expenditure. Within each category, studies were found for training effects and for acute effects. Category 1: Several small, randomized controlled trials showed that improvements in measures of cardiorespiratory fitness did not differ significantly between training regimens based on long or short sessions. Acute effects of two short sessions on excess postexercise oxygen consumption were reported to be greater than those of one longer session. By contrast, short-term decreases in postprandial triglyceride concentrations were found to be similar with three short or one long session. Category 2: Higher-intensity training was consistently found to elicit greater increases in VO(2max) than lower-intensity training of longer duration. No conclusion could be drawn for any other outcome. A session of hard/moderate exercise may be more likely than to induce short-term negative energy balance than light exercise. Findings on the comparability of sessions of different intensities on blood lipids and glucose/insulin dynamics are conflicting. CONCLUSION: Further research is required before the principle of fractionization can be endorsed with confidence. PMID- 11427767 TI - Dose response between total volume of physical activity and health and fitness. AB - PURPOSE: Studies published in 1990s were evaluated for the possible dose response between the total volume of physical activity and the fitness and health outcomes, and for the characteristics of the dose response relations. METHODS: Nineteen observational studies and 15 randomized trials were identified. The scope of the studies was on primary prevention among inactive, healthy, middle aged and elderly men and women. MET-min x wk(-1) was used as the primary volume measure. No studies addressing specifically the volume-outcome dose response were identified. RESULTS: The cross-sectional and follow-up studies suggested a graded dose response of the volume of physical activity with all-cause mortality, stroke and several coronary heart disease risk factors. The benefits were apparent among both men and women. Nonrandomized and uncontrolled randomized trials exhibited no clear dose response relationship, whereas the randomized controlled trials showed a crude graded dose response between the exercise volume as measured by MET-min x wk(-1) and VO(2max) but not between volume and disease risk factors. An apparently clearer dose response was seen between the intensity of physical activity and the VO(2max) response. These data do not allow for quantitative characterization of the observed dose response relations between physical activity volume and health and fitness. CONCLUSION: Fairly strong evidence indicates a crude dose response between the total volume of weekly physical activity and cardiorespiratory fitness but only weak evidence for a dose response of activity volume and health measurers. PMID- 11427768 TI - The acute versus the chronic response to exercise. AB - PURPOSE: There is strong and consistent evidence that a single exercise session can acutely reduce triglycerides and increase high-density lipoprotein (HDL) cholesterol (HDL-C), reduce blood pressure, and improve insulin sensitivity and glucose homeostasis. Such observations suggest that at least some of the effects on atherosclerotic cardiovascular disease (ASCVD) risk factors attributed to exercise training may be the result of recent exercise. RESULTS: These acute and chronic exercise effects cannot be considered in isolation. Exercise training increases the capacity for exercise, thereby permitting more vigorous and/or more prolonged individual exercise sessions and a more significant acute effect. The intensity, duration, and energy expenditure required to produce these acute exercise effects are not clearly defined. The acute effect of exercise on triglycerides and HDL-C appears to increase with overall energy expenditure possibly because the effect maybe mediated by reductions in intramuscular triglycerides. Prolonged exercise appears necessary for an acute effect of exercise on low-density lipoprotein (LDL) cholesterol (LDL-C) levels. The acute effect of exercise on blood pressure is a low threshold phenomenon and has been observed after energy expenditures requiring only 40% maximal capacity. The acute effect of exercise on glucose metabolism appears to require exercise near 70% maximal, but this issue has not been carefully examined. CONCLUSIONS: Exercise has definite acute effects on blood lipids, blood pressure, and glucose homeostasis. Exercise also has acute effects on other factors related to atherosclerosis such as immunological function, vascular reactivity, and hemostasis. Considerable additional research is required to define the threshold of exercise required to produce these putatively beneficial effects. PMID- 11427769 TI - Individual differences in response to regular physical activity. AB - PURPOSE: The purpose of this review was to address the question of interindividual variation in responsiveness to regular exercise training and to define the contributions of age, sex, race, and pretraining phenotype level to this variability. METHODS: A literature review was conducted of the studies reporting interindividual variation in responsiveness to standardized and controlled exercise-training programs, and included an analysis of the contribution of age, sex, race, and initial phenotype values to the heterogeneity in VO(2max), high-density lipoprotein (HDL)-C and submaximal exercise, heart rate (HR), and systolic blood pressure (SBP) training responses in subjects from the HERITAGE Family Study. RESULTS: Several studies have shown marked individual differences in responsiveness to exercise training. For example, VO(2max) responses to standardized training programs have ranged from almost no gain up to 100% increase in large groups of sedentary individuals. A similar pattern of heterogeneity has been observed for other phenotypes. Data from the HERITAGE Family Study show that age, sex, and race have little impact on interindividual differences in training responses. On the other hand, the initial level of a phenotype is a major determinant of training response for some traits, such as submaximal exercise heart rate and blood pressure (BP) but has only a minor effect on others (e.g., VO(2max), HDL-C). The contribution of familial factors (shared environment and genetic factors) is supported by data on significant familial aggregation of training response phenotypes. CONCLUSIONS: There is strong evidence for considerable heterogeneity in the responsiveness to regular physical activity. Age, sex, and ethnic origin are not major determinants of human responses to regular physical activity, whereas the pretraining level of a phenotype has a considerable impact in some cases. Familial factors also contribute significantly to variability in training response. PMID- 11427771 TI - What to look for in assessing responsiveness to exercise in a health context. AB - PURPOSE: When attempting to assess responsiveness to habitual exercise in a health context, there is a wide range of issues that need to be addressed in order to provide science-based recommendations for use in evidence-informed health care delivery. Issues regarding responsiveness include characteristics of the exercise regimen or dose as well as characteristics of the response or effect. RESULTS: The exercise dose typically has been characterized by its type, intensity, session duration, and session frequency with the concepts of activity profile, activity volume, and accumulation over multiple bouts throughout the day recently added to these discussions. When establishing the dose for a designated outcome, specificity of the response in relation to exercise type needs to be considered. Understanding the role of intensity as a stimulus for change is critical because of the intensity-related biological responses to exercise, its role in exercise-induce medical complications, and the aversion many adults have to vigorous exercise. Receiving considerable attention is whether a higher intensity or greater volume of endurance-type exercise is more important in producing a variety of health-related benefits. To understand the potential role of accumulation of exercise, more information is needed on benefits derived from very short (< or = 5 min) bouts of exercise performed frequently as well as very long bouts (> or = 90 min) performed infrequently. CONCLUSIONS: When considering the response, it is important to establish the priority health outcomes, the relationship of the dose response for individual biological variables to the dose response for clinical outcomes, the basis for substantial interindividual variations in the response to a specific exercise dose, and the health benefit to health risk relationship for various doses of exercise. Scientific resolution of these issues will substantially facilitate the development and dissemination of appropriate guidelines for the use of exercise in promoting health. PMID- 11427772 TI - Physical activity and all-cause mortality: what is the dose-response relation? AB - PURPOSE: The purpose of this review is to assess the dose-response relation between physical activity and all-cause mortality. We examined these parameters of physical activity dose: volume, intensity, duration, and frequency. METHODS: We used a computer-assisted literature search to identify papers on this topic. After excluding papers examining only two levels of physical activity or fitness, papers investigating specific causes of mortality, reviews, and those not written in English, 44 papers satisfying all criteria were included in this review. RESULTS: There is clear evidence of an inverse linear dose-response relation between volume of physical activity and all-cause mortality rates in men and women, and in younger and older (> or = 60 yr) persons. Minimal adherence to current physical activity guidelines, which yield an energy expenditure of about 1000 kcal x wk(-1) (4200 kJ x wk(-1)), is associated with a significant 20--30% reduction in risk of all-cause mortality. Further reductions in risk are observed at higher volumes of energy expenditure. It is unclear whether a volume of <1000 kcal x wk(-1) also may be associated with lower risk; there are some data supporting this. Due to limited data, it is also unclear whether vigorous intensity activity confers additional benefit beyond its contribution to volume of physical activity when compared with moderate-intensity activity. No data are available on duration and frequency of physical activity in relation to all-cause mortality rates after controlling for volume of physical activity. CONCLUSIONS: All studies in this review are observational studies, so conclusions are based on Evidence Category C. There is an inverse linear dose-response relation between volume of physical activity and all-cause mortality. Further research is needed to clarify the contributions of its components--intensity, duration, and frequency--to decreased all-cause mortality rates. PMID- 11427773 TI - Physical activity and cardiovascular disease: evidence for a dose response. AB - PURPOSE: To summarize and synthesize existing literature providing evidence of a dose-response relation between physical activity and cardiovascular disease endpoints. METHODS: MEDLINE search of indexed English-language literature through August 2000. Findings supplemented by existing consensus documents and other published literature. Only studies with greater than two physical activity exposure categories were included, and studies not focusing on the clinical manifestation of the outcome (incidence or mortality) were excluded. RESULTS: Existing studies were classified by outcome used: all cardiovascular disease (CVD), coronary (ischemic) heart disease (CHD), and stroke. The vast majority of the literature in this area has relied on prospective observational studies and has been conducted in European men or populations of men of primarily European descent. Follow-up intervals ranged from 3 to 26 yr, and most studies related a single initial measure of physical activity to the outcome of interest, sometimes many years in the future. No randomized trials of physical activity and cardiovascular disease as a clinical outcome exist. Taken together, the available evidence indicates that cardiovascular disease incidence and mortality, and specifically ischemic heart disease, are causally related to physical activity in an inverse, dose-response fashion. These findings have been demonstrated in a variety of populations and using a variety of physical activity assessment methods. Contrarily, equivocal evidence for stroke incidence and mortality prohibits a similar conclusion. No strong evidence for dose-response relation between physical activity and stroke as a CVD outcome is available. CONCLUSION: Physical inactivity is prominent in the causal constellation for factors predisposing to cardiovascular disease, particularly ischemic heart disease. Methodologic advances in physical activity assessment; additional studies on changes in the antecedent variable, physical activity, as it relates to the outcome; and more studies among women and ethnically diverse populations are needed to clarify these relations. PMID- 11427774 TI - Exercise characteristics and the blood pressure response to dynamic physical training. AB - PURPOSE: The purpose of this study was to assess the influence of the characteristics of the exercise program, particularly exercise intensity, on the blood pressure response to dynamic physical training in otherwise healthy normotensive and hypertensive subjects. METHODS: This study is a meta-analysis of randomized controlled intervention trials and a description of studies in which different training regimens have been compared. RESULTS: The weighted net reduction of blood pressure in response to dynamic physical training averaged 3.4/2.4 mm Hg (P < 0.001). Interstudy differences in the changes in pressure were not related to weekly frequency, time per session, or exercise intensity, which ranged from approximately 45--85%; these three characteristics combined explained less than 5% of the variance of the blood pressure response. The response of diastolic blood pressure was not different according to training intensity in studies that randomized patients to training programs with different intensities. Some studies reported a greater reduction of systolic blood pressure when intensity was about 40% than when participants exercised at about 70%, but this finding was not consistent, neither within nor between studies. CONCLUSION: Training from three to five times per week during 30--60 min per session at an intensity of about 40--50% of net maximal exercise performance appears to be effective with regard to blood pressure reduction. The evidence that higher intensity exercise would be less effective is at present inconsistent. PMID- 11427776 TI - Effects of exercise on glucose homeostasis in Type 2 diabetes mellitus. AB - PURPOSE: This review has evaluated the effectiveness of physical activity in the treatment and prevention of Type 2 diabetes mellitus (DM). METHODS: The available literature was reviewed through a computerized search (MEDLINE, 1966--2000) to classify studies examining the influence of physical activity on the treatment and prevention of Type 2 DM. Additional studies were reviewed through ancestral searches from their bibliographies. RESULTS: It is not possible at the present time to discern from the available literature whether a dose-response relationship exists between exercise volume or intensity and improvements in glucose control in Type 2 DM. Large-scale, prospective studies indicate that higher levels of physical activity are clearly associated with a lower incidence of Type 2 DM. However, no randomized controlled trials have been conducted to address the dose-response effect of exercise or physical activity on diabetes treatment or prevention. CONCLUSION: It is uncertain whether there is a dose response effect of exercise on improved glucose control in Type 2 DM. There does appear to be, however, a limited amount of evidence suggesting that increasing levels of physical activity contribute to better diabetes prevention. Clearly, additional studies are needed to determine the influence of physical activity on the treatment and prevention of Type 2 DM. PMID- 11427777 TI - Response of blood lipids to exercise training alone or combined with dietary intervention. AB - PURPOSE: The purpose of this study is to review the effects of aerobic exercise training (AET) on blood lipids and assess dose-response relationships and diet interactions. METHODS: We reviewed papers published over the past three decades pertaining to intervention trials on the effects of > or = 12 wk of AET on blood lipids and lipoprotein outcomes in adult men and women. Included were studies with simultaneous dietary and AET interventions, if they had appropriate comparison groups. Studies were classified by the participants' relative weights expressed as mean BMIs. Information was extracted on baseline characteristics of study subjects, including age, sex, and relative baseline cholesterol levels; details on the training programs; and the responses to training of body weight, VO(2max), and blood total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C), and triglyceride (TG). RESULTS: We identified 51 studies, 28 of which were randomized controlled trials. AET was generally performed at a moderate to hard intensity, with weekly energy expenditures ranging from 2,090 to >20,000 kJ. A marked inconsistency was observed in responsiveness of blood lipids. The most commonly observed change was an increase in HDL-C (with reductions in TC, LDL-C, and TG less frequently observed). Insufficient data are available to establish dose response relationships between exercise intensity and volume with lipid changes. The increase in HDL-C with AET was inversely associated with its baseline level (r = -0.462), but no significant associations were found with age, sex, weekly volume of exercise, or with exercise-induced changes in body weight or VO(2max). CONCLUSION: Moderate- to hard-intensity AET inconsistently results in an improvement in the blood lipid profile, with the data insufficient to establish dose-response relationships. PMID- 11427778 TI - Dose-response and coagulation and hemostatic factors. AB - PURPOSE: The purpose of this study was to evaluate the dose-response relations of regular physical activity on platelet function, blood coagulation factors, and fibrinolytic factors, on the basis of studies with appropriate experimental design. METHODS: MEDLINE-based literature search supplemented with relevant review articles and other individual articles was used. The review concentrates on controlled randomized clinical trials on the impact of regular physical exercise on platelet function, and on blood coagulation and fibrinolytic factors. RESULTS: Physical exercise acutely activates platelets and the fibrinolytic system, and some factors of the blood coagulation cascade. These findings, the earliest of which were published already in the 1960s, are mainly derived from uncontrolled observations. These studies have led to the conclusion that unbalanced activation of the hemostatic system during acute exercise may be part of the mechanisms for sudden cardiac events during or shortly after heavy physical exercise. The effects of regular physical exercise on various aspects of platelet function, blood coagulation, and fibrinolysis have been the object in only a few controlled randomized trials. With the exception of data on platelet function, the results remain contradictory. CONCLUSION: Controlled randomized clinical trials with adequate statistical power and experimental study designs with subjects of different ages and health status are warranted for the evaluation of the dose-response issues to clearly delineate the preventive and therapeutic potential of regular physical exercise. PMID- 11427779 TI - Physical activity, total and regional obesity: dose-response considerations. AB - PURPOSE: This review was undertaken to determine whether exercise-induced weight loss was associated with corresponding reductions in total, abdominal, and visceral fat in a dose-response manner. METHODS: A literature search (MEDLINE, 1966--2000) was performed using appropriate keywords to identify studies that consider the influence of exercise-induced weight loss on total and/or abdominal fat. The reference lists of those studies identified were cross-referenced for additional studies. RESULTS: Total fat. Review of available evidence suggested that studies evaluating the utility of physical activity as a means of obesity reduction could be subdivided into two categories based on study duration. Short term studies (< or = 16 wk, N = 20) were characterized by exercise programs that increased energy expenditure by values double (2200 vs 1100 kcal.wk-1) that of long-term studies (> or = 26 wk, N = 11). Accordingly, short-term studies report reductions in body weight (-0.18 vs -0.06 kg x wk(-1)) and total fat (-0.21 vs 0.06 kg x wk(-1)) that are threefold higher than those reported in long-term studies. Moreover, with respect to dose-response issues, the evidence from short term studies suggest that exercise-induced weight loss is positively related to reductions in total fat in a dose-response manner. No such relationship was observed when the results from long-term studies were examined. Abdominal fat. Limited evidence suggests that exercise-induced weight loss is associated with reductions in abdominal obesity as measured by waist circumference or imaging methods; however, at present there is insufficient evidence to determine a dose response relationship between physical activity, and abdominal or visceral fat. CONCLUSION: In response to well-controlled, short-term trials, increasing physical activity expressed as energy expended per week is positively related to reductions in total adiposity in a dose-response manner. Although physical activity is associated with reduction in abdominal and visceral fat, there is insufficient evidence to determine a dose-response relationship. PMID- 11427781 TI - Physical activity and cancer risk: dose-response and cancer, all sites and site specific. AB - PURPOSE: The association between physical activity and overall and site-specific cancer risk is elaborated in relation to whether any observed dose-response association between physical activity and cancer can be interpreted in terms of how much physical activity (type, intensity, duration, frequency) is needed to influence site- and gender-specific cancer risk. METHODS: Observational studies were reviewed that have examined the independent effect of the volume of occupational physical activity (OPA) and/or leisure time physical activity (LPA) on overall and site-specific cancer risk. RESULTS: The evidence of cohort and case-control studies suggests that both leisure time and occupational physical activity protect against overall cancer risk, with a graded dose-response association suggested in both sexes. Confounding effects such as diet, body weight, and parity are often included as a covariate in the analyses, with little influence on the observed associations. A crude graded inverse dose-response association was observed between physical activity and colon cancer in 48 studies including 40,674 colon/colorectal cancer cases for both sexes. A dose-response effect of physical activity on colon cancer risk was especially observed, when participation in activities of at least moderate activity (>4.5 MET) and demonstrated by activities expressed as MET-hours per week. An observed inverse association with a dose-response relationship between physical activity and breast cancer was also identified in the majority of the 41 studies including 108,031 breast cancer cases. The dose-response relationship was in particular observed in case-control studies and supported by observations in cohort studies when participation in activities of at least moderate activity (>4.5 MET) and demonstrated by activities expressed by MET-hours per week. This association between physical activity and breast cancer risk is possibly dependent on age at exposure, age at diagnosis, menopausal status and other effect modifiers, e.g., body mass index. Furthermore, data concerning carcinoma of other cancers (prostate, lung, endometrium, ovary, and testicular cancers) are required. CONCLUSION: A protective effect of physical activity on site-specific cancer risk with a dose-response association between physical activity and colon and pre- and postmenopausal breast cancer supported by identified biological mechanisms has been observed. The optimal permutation of type, intensity, duration, and frequency of physical activity across the lifespan is unclear, but it is gender, age, and site specific and supports moderate activity (>4.5 MET) more than light activities (<4.5 MET). The complicated nature of the physical activity variable, combined with lack of knowledge regarding possible biological mechanisms operating between physical activity and cancer, warrants further studies including controlled clinical randomized trials. PMID- 11427782 TI - Dose-response of physical activity and low back pain, osteoarthritis, and osteoporosis. AB - PURPOSE: The purpose of this study was to examine the evidence for causal relationships between physical activity (PA) and low back pain (LBP), osteoarthritis (OA), and osteoporosis (OP), and for dose-response relations involved. METHODS: Computer database searches and personal retrieval systems were used to locate relevant literature. RESULTS: PA can be effective in preventing LBP (Category A) but prolonged, heavy loading can lead to LBP (Category C). Specific exercises have not been found effective in treatment of acute LBP (Category A), but PA can be effective in chronic LBP (Category B), especially for diminishing the effects of deconditioning. No evidence indicates that PA directly prevents OA. Large amounts of intensive PA involving high impacts or torsional loadings or causing injuries increases risk of OA (Category C). Light or moderate PA does not increase the risk of OA (Category C). PA can be effective in the treatment and rehabilitation of OA (Category B). High-intensity loading is osteogenic and possibly useful in prevention of OP (Category A) at the loaded site, but low to moderate loading is not osteogenic (Category D). Static efforts and slow movements are ineffective or less effective than fast application of force (Category B). The types of PA to attain the effects mentioned above are known except in the case of prevention of LBP, but dose-response relationships are poorly known; at best, semiquantitatively on the basis of just a few studies. CONCLUSION: Given the shown primary and/or secondary preventative effectiveness of PA regarding LBP, OA, and OP, research to elucidate the inadequately known dose-response relations should be given high priority. PMID- 11427783 TI - Physical activity dose-response effects on outcomes of depression and anxiety. AB - PURPOSE: The purpose of this study was to examine the scientific evidence for a dose-response relation of physical activity with depressive and anxiety disorders. METHODS: Computer database searches of MEDLINE, PsychLit, and Internet and personal retrieval systems to locate population studies, randomized controlled trials (RCTs), observational studies, and consensus panel judgments were conducted. RESULTS: Observational studies demonstrate that greater amounts of occupational and leisure time physical activity are generally associated with reduced symptoms of depression. Quasi-experimental studies show that light-, moderate-, and vigorous-intensity exercise can reduce symptoms of depression. However, no RCTs have varied frequency or duration of exercise and controlled for total energy expenditure in studies of depression or anxiety. Quasi-experimental and RCTs demonstrate that both resistance training and aerobic exercise can reduce symptoms of depression. Finally, the relation of exercise dose to changes in cardiorespiratory fitness is equivocal with some studies showing that fitness is associated with reduction of symptoms and others that have demonstrated reduction in symptoms without increases in fitness. CONCLUSION: All evidence for dose-response effects of physical activity and exercise come from B and C levels of evidence. There is little evidence for dose-response effects, though this is largely because of a lack of studies rather than a lack of evidence. A dose response relation does, however, remain plausible. PMID- 11427784 TI - Exercise dose-response effects on quality of life and independent living in older adults. AB - PURPOSE: The purpose of this study was to determine if exercise operates in a dose-response fashion to influence well-being and to postpone dependency. METHODS: A computer-assisted search was made by using the following key words: resistance training, strength training, function, exercise, elderly, quality of life, frailty, physical activity, independence, performance, aerobic training, mobility, well-being, and disability. Review articles and personal files were also used, and a critical review of research studies meeting the criteria described in the methods section of the article was conducted. RESULTS: In large sample correlational studies and prospective longitudinal studies, researchers consistently report that measures of physical function in old adults are related to feelings of well-being, and that old adults who are physically active also report higher levels of well-being and physical function, but the results of randomized intervention studies of aerobic and/or resistive strength training do not always support this relationship. Even if changes in well-being and physical function were reported, no evidence was found that levels of intensity operated in a dose-response fashion to influence these changes. Research design problems included ineffective aerobic or strength training treatments, widely varying participation and effort of the research participants, and both treatment and physical function tests that were not appropriate for the physical status of the participants. CONCLUSION: The most consistent results were that long-term physical activity is related to postponed disability and independent living in the oldest-old subjects. Even in individuals with chronic disease, systematic participation in physical activities enhances physical function. PMID- 11427786 TI - Health effects resulting from exercise versus those from body fat loss. AB - OBJECTIVE: The purpose of this review was to assess whether body weight confounds the relationships between physical activity and its health benefits. METHODS: The review includes 80 reports from population-based studies (Evidence Category C) of physical activity or fitness and cardiovascular disease (CVD) or coronary heart disease (CHD). RESULTS: Eleven of 64 reports on activity found no relationship between physical activity and disease. Of the remaining 53 reports, 11 did not address the possible confounding effects of body weight, nine cited reasons that weight differences should not explain their observed associations, and 33 statistically adjusted for weight (as required). Only three of these changed their associations from significant to nonsignificant when adjusted. Ten of 16 reports on cardiorespiratory fitness and CHD or CVD used statistical adjustment, and none of these changed their findings to nonsignificant. Population studies show that vigorously active individuals also have higher high-density lipoprotein (HDL)-cholesterol concentration, a major risk factor for CHD and CVD, than sedentary individuals when statistically adjusted for weight. In contrast, intervention studies, which relate dynamic changes in weight and HDL, suggest that adjustment for weight loss largely eliminates the increase in HDL cholesterol in sedentary men who begin exercising vigorously. Adjusting the cross sectional HDL-cholesterol differences for the dynamic effects of weight loss eliminates most of the HDL-cholesterol difference between active and sedentary men. CONCLUSION: Population studies show that the lower incidence of CHD and CVD and higher HDL of fit, active individuals are not because of lean, healthy individuals choosing to be active (i.e., self-selection bias). Nevertheless, metabolic processed associated weight loss may be primarily responsible for the HDL differences between active and sedentary men, and possibly also their differences in CHD and CVD. PMID- 11427787 TI - Dose-response: variation with age, sex, and health status. AB - PURPOSE: The purpose of this study was to summarize the literature on the influence of age, sex, and health status on the changes in systolic (SBP) and diastolic blood pressure (DBP), triglycerides (TG), and high-density lipoprotein cholesterol (HDL-C) consequent to exercise training. METHODS: A MEDLINE search was performed from January 1966 through August 2000 to identify studies that have investigated the effects of age, sex, and health status on the changes in the outcome variables with exercise training. References from these studies and from review and meta-analysis studies were also reviewed. RESULTS AND CONCLUSIONS: The results indicate that age has little or no influence on the changes in SBP, DBP, TG, and HDL-C in response to exercise training. When looking at sex, females appear to have an attenuated response to exercise training compared with males with respect to SBP, DBP, and HDL-C, but the data for TG are equivocal. Finally, there appears to be more favorable changes in resting SBP and DBP, TG, and HDL-C in unhealthy subjects (hypertensive and post-MI patients) when compared with healthy subjects. PMID- 11427788 TI - Population attributable risk: implications of physical activity dose. AB - PURPOSE: The purpose of this study is to describe the application of population attributable risk estimates in relation to the dose-related benefits or risks of physical activity. METHODS: Assumptions and limitations of population attributable risk calculations and interpretations are reviewed and evaluated in the context of physical activity dose. Theoretical estimates are developed for several hypothetical situations. RESULTS: National estimates of population attributable risk may be inaccurate because definitions and measurement techniques applied in physical activity research studies and physical activity prevalence surveys do not correspond. In addition, it is not established whether vigorous or moderate physical activity are independent contributors, sequential categories, or interactive variables in the process of disease reduction. This information is necessary to calculate population attributable risk most appropriately. CONCLUSION: Estimates of the disease burden of physical inactivity will be improved by two advances in empirical studies: first, the pairing of prevalence and relative risk estimates for nationally representative population based samples; and second, refined relative risk estimates for various doses of physical activity. PMID- 11427791 TI - [Diagnostic criteria: for whom? Why? Which tests?]. PMID- 11427792 TI - [Role of the hemochromatosis gene in prophyria cutanea tarda. Prospective study of 56 cases]. AB - BACKGROUND: The cause of iron overload in prophyria cutanea tada is unknown. The aim of this work was to determine the frequency of the hemochromatosis gene (HFE) in 56 patients with porphyria cutanea tarda. We analyzed the relationship between HFE mutations and biochemical abnormalities in porphyria cutanea tarda and the interaction with other triggering factors of porphyria cutanea tarda (alcohol abuse, hepatitis C, drugs). PATIENTS AND METHODS: Hepatitis C, alcohol abuse, drug intake and HFE mutations were determined in 56 patients with porphyria cutanea tarda (44 men and 12 women). Iron status was determined from transferrin saturation, serum iron, and serum ferritin. Liver metabolism was determined from liver chemistries: alanine aminotransferase, aspartate aminotransferase, and gamma-glutamyl transpeptidase. RESULTS: Thirty-nine patients (69.4 p. 100) carried HFE mutations, 18 (32.1 p. 100) were H63D heterozygous, 4 (7.1 p. 100) were H63D homozygous, 9 (16 p. 100) C282Y heterozygous, 8 (14.2 p. 100) compound C282Y/H63D heterozygous and none were C282Y homozygous. Comparison between porphyria cutanea tarda with and without mutations showed that compound C282Y/H63D heterozygous status was significantly linked to iron overload: transferrin saturation=0.61 vs 0.39 (p=0.0001) and serum iron=32.9 vs 22.4 (p=0.0046). H63D homozygous status was linked to iron overload but non significantly: transferrin sturatin=0.53 vs 0.39 (p=0.06). The class with high iron overload (transferrin saturation > 0.45) was not linked with triggering factors of porphyria cutanea tarda. Hepatatic cytolysis was linked to alcohol abuse and hepatitis C but not to HFE mutations. DISCUSSION: The frequencies of HFE mutations in Lyons France are halfway between Anglo-Saxon and Italian papers, highlighting the Celtic origin of C282Y mutation. Compound heterozygous and to a lesser degree H63D homozygous status explained the highest iron overload in our patients. This favors clinical expression of porphyria cutanea tarda. This iron overload due to HFE mutations is a new triggering factor of porphyria cutanea tarda independent of classical triggering factors: mutation of the erythrocytic uroporpyrinogen decarbocylase gene, alcohol abuse, hepatitis C, and drugs. PMID- 11427793 TI - [Pityriasis rosea in secondary schools in Ouagadougou, Burkina Faso]. AB - BACKGROUND: Pityriasis rosea is a spontaneously regressive benign erythematous squamous dermatosis. A better understanding of this condition in developing countries would be useful. PATIENTS AND METHODS: We conducted a cross section study based on one-day surveys in Ouagadougou, Burkina Faso, secondary schools. The stratified cluster sampling method described by Henderson was used. Six thousand pupils were examined to collect epidemiology and clinical data and to determine the presence of complications and results of complementary explorations performed. RESULTS: Thirty-six cases of pityriasis rosea were observed (0.6 p. 100). Prevalence was higher in children from an unfavorable social and economic background. A typical eruption was often observed with classical localizations and morphology in 86.1 p. 100 of the cases. Pruritus was often observed with an inaugural lesion (61.1 p. 100) predominantly on the upper limbs (54.6 p. 100) and the trunk (31.8 p. 100) measuring 1 to 3 cm. A typical second eruption was found in most cases, situated on the trunk (75 p. 100), distal portions of the limbs (64.4 p. 100), proximal portions of the limbs (33.3 p. 100) and the neck (30.5 p. 100). The eruption evolved for 1 week to 1 month in most cases (61 p. 100). In 55.5 p. 100 of the cases, therapeutic abstention was the rule. Secondary treatment-related complications occurred in 38.9 p. 100 of the cases. Stool studies generally demonstrate ameba cysts. Syphilis serology was negative. DISCUSSION: Our sampling technique in the secondary school setting provided a representative sample of the urban population of African cities. This work demonstrated the variability of prevalence by social and economic conditions and is in favor of an infections cause, demonstrating several similar features with other series reported in the literature. Generally, a stool study is not necessary and syphilis serology is negative in typical cases. Health care workers should be more aware of pityriasis rosea in order to avoid poorly adapted treatment. PMID- 11427794 TI - [Sarcoidosis: thalidomide treatment in ten patients]. AB - BACKGROUND: Acute cutaneous sarcoidosis is generally spontaneously regressive but persistent chronic cutaneous lesions are esthetically prejudicial. There have been several case reports on thalidomide efficacy but long-term outcome is unknown. We report results in 10 cases of cutaneous sarcoidosis treated with thalidomide. PATIENTS AND METHODS: Data from ten patients with sarcoidosis treated with thalidomide between January 1998 and March 1999 were collected from delivery authorizations and analyzed. All ten patients had chronic cutaneous sarcoidosis resistant to conventional therapy. Six patients had an associated visceral localization and disease duration of 2 to 18 years (median 6 years). We considered that regression was complete when erythema and infiltration had totally disappeared, that regression was incomplete when cutaneous signs remained, and that treatment had failed when no effect was observed or when the disease worsened. RESULTS: Disease regression was noted in 7 patients for a daily dose of 1.84 mg/kg for 2.8 months. Skin lesions totally regressed in 3 patients, an incompletely in 4. Treatment failed in 3 patients. Patients were treated for 10 months (2 to 21 months). The daily dose of thalidomide was gradually reduced in 5 of 7 patients for whom treatment was effective. Three of these 5 patients relapsed and thalidomide was again given and was effective again at the same dose and after the same delay. We observed improved kidney function in one patient, improvement in nasal infiltration in one other and complete regression in 3 patients who achieved long lasting reduction in angiotensin convertase level. When treatment failed, the daily dose was 1.15 mg/kg and the treatment had to be stopped for 2 patients. Side effects were minor, excepting 2 cases of neuropathy. DISCUSSION: This open study of 10 patients treated with thalidomide showed the efficacy of a 1.84 mg/kg daily dose in 7 out of 10, but complete regression of the lesions was obtained in only 3 patients. Thalidomiide appears to suspend the disease, with relapse when the drug is discontinued and efficacy at re introduction. This would argue against a placebo effect. The mode of action could involve immunomodulating and antiinflammatory mechanisms. PMID- 11427795 TI - [Sensitization to resorcinol in a prescription verrucide preparation: unusual systemic clinical features and prevalence]. AB - BACKGROUND: Resorcinol is a rare sensitizer. In Lorraine, where it is used in high concentrations in an anti-wart ointment, this molecule has induced many cases of allergy. The purpose of this study was to describe clinical features in patients sensitized to resorcinol, to determine the incidence of sensitization in Lorraine and ascertain the occurrence of cross reactions with other phenolic pro haptens. PATIENTS AND METHODS: The files of all patients sensitized to resorcinol were studied from 1992 to 1999. Over a 20-month period, 983 consecutive patients underwent our standard patch tests where we included resorcinol (1 p. 100 in petrolatum). Co-sensitization with other pro-haptens was studied in all sensitized patients. RESULTS: Sensitization to resorcinol was observed in 0.5 p. 100 of the 983 consecutively tested patients. Contact sensitization was found in 24 patients, who all but one had previously used the anti-wart ointment containing resorcinol. All developed contact eczema on the site of application of the ointment, with generalized urticaria (4 cases), pompholyx (1 case), and generalized papulo-vesicular rash with pompholyx (6 cases). Positive patch tests were observed with resorcinol monobenzoate (15/19 cases), pyrocatechol (7/14 cases), pyrogallol (9/19 cases), salicylaldehyde (2/17 cases), and hydroquinone (6/17 cases). Negative patch tests were observed with phenysalicylate, hexylresorcinol, or fluoresceine. CONCLUSION: Resorcinol should not be applied in high concentrations as it can provoke generalized sensitization and co sensitization with other pro-haptens. PMID- 11427796 TI - [Large hairy pigmented spots in neurofibromatosis type 1: an atypical form of neurofibromas]. AB - INTRODUCTION: Large hairy pigmented spots have been observed in patients with neurofibromatosis type 1. In this study we tried to determine the nature and the frequency of these hairy pigmented spots in neurofibromatosis type 1. PATIENTS AND METHODS: In patients with neurofibromatosis type 1, hairy pigmented spots with a diameter more than 3 cm were systematically notified. Realisation of the biopsy of the spot was proposed to the patient. RESULTS: Among 614 patients with neurofibromatosis type 1, seven (1.1 p. 100) had a large hairy pigmented spot. Biopsy was realized in six cases. In five cases, diagnosis was superficial and plexiform neurofibroma, the 6(th) case was a Becker's nevus. CONCLUSION: Large hairy pigmented spot is a rare aspect of superficial and plexiform neurofibroma during neurofibromatosis type 1. A biopsy may be useful if it is necessary for the disorder diagnosis. PMID- 11427797 TI - [Atopic dermatitis in Tunisia: epidemiological and clinical aspects]. AB - INTRODUCTION: The prevalence of atopic dermatitis is usually high in western countries, varying from 18 p. 100 to 20 p. 100. Recent studies suggest an increasing of this frequency. The aim of our study is to determine the epidemiological and clinical characteristics of atopic dermatitis in Tunisia through the analysis of a retrospective cohort. PATIENTS AND METHODS: We retrospectively studied all the medical reports of atopic dermatitis registrated in the department of dermatology during a 7 years period (1992-1998). We analysed epidemiological and clinical features of every medical report. We used diagnosis criteria of Hanifin and Rajka. RESULTS: Four hundred fifty-one cases of atopic dermatitis have been diagnosed (54 adults and 397 infants). Relative frequency of new cases of atopic dermatitis compared to new diagnosis was 0.37 p. 100 in 1992 and 0.72 p. 100 in 1998. Mean age was 3.2 years. A personal history of atopy was observed for 15.2 p. 100 of patients and a family history of atopy for 35.9 p. 100. A few numbers of complications were observed. Bacterial infections interested 15.3 p. 100 of cases. Hospitalisation was needed for 4 patients (0.88 p. 100). Class III and IV topical corticosteroids were usually used (84.8 p. 100). Potent corticosteroids (class I) were required for only 3.2 p. 100 of cases. CONCLUSIONS: We notice a low frequency of atopic dermatitis in this study and a predominance of mild forms of the disease. Other studies are needed to confirm these results and to determine the prevalence of atopic dermatitis in Tunisia. PMID- 11427798 TI - [Minocycline]. AB - Minocycline belongs to the second generation class of cyclines. It was synthesized in 1967 and marketed in 1972. Minocycline has an antiinfectious activity with a spectrum similar to that of other cyclines, notably against Chlamydias, Treonema and Proprionibacterium acenes. The antiinflammatory activity is associated with this antiinfectious action is greater than that of first generation cyclines with specifically a modulator effect on epidermal cytokines. The pharmokinetics of minocycline is characterized by an excellent absorption, a long half-life and an important lipophilic property inducing good tissue distribution. Clinical trials of minocycline have mainly been performed in sexually transmissible diseases and in acne, a field where randomized studies are the most frequent. These trials show that the effect of minocycline is not stronger than first generation cyclines or doxycycline, but that the action is quicker than that of tetracycline at the dose of 500 mg a day. Minocycline is also efficient in nocardiasis, mycobacteriosis, leprosy, Lyme disease, pyoderma gangrenosum, autoimmune bullous dermatitis, Carteaud disease, and prurigo. However, the effect of minocycline in these different conditions has always been evaluated in open trials with a small number of patients. The usual side effects of cyclines, i.e. digestive problems, fungal infections, are less frequent than with first generation cyclines. No photosensitivity has been demonstrated although pigmentations have been described. Dizziness is a specific side effect of minocycline. Furthermore, rare but severe side effects have been reported, including hypersensitivity syndrome, autoimmune hepatitis, and lupus. Regular indications for minocycline in dermatology are acne and three sexually transmissible diseases (mycoplasm, chlamydia, treponema). Proposed dosage is 100 mg per day in sexually transmissible disease with a reduction to 50 mg per day after 15 days in acne. PMID- 11427799 TI - [Pemphigoid gestationis: treatment by topical class I corticosteroid]. AB - BACKGROUND: Systemic corticosteroid is the main treatment of severe forms of pemphigoid gestationis. We report a case of generalised pemphigoid gestationis successfully treated with very potent topical corticosteroid. CASE: A 37-year-old woman developed during her third pregnancy with a new partner an urticated generalised eruption associated with bullous lesions. The diagnosis of pemphigoid gestationis was confirmed by direct immunofluorescence which detected a linear C3 deposition along the basement membrane zone and the positivity of Herpes Gestationis Factor (10 units). Local treatment with potent corticosteroid (betamethasone dipropionate 0.05 p. 100) failed and the patient was successfully treated by clobetasol propionate 0.05 p. 100 cream. The infant, in good health, was not delivered prematurely. DISCUSSION: Severe form of pemphigoid gestationis are currently treated with 0.5 to 1 mg/kg/day of systemic corticosteroids, with maternal and pediatric possible side effects. As in bullous pemphigoid, this observation underlines the efficiency and good tolerance of very potent corticosteroid in severe forms of pemphigoid gestationis. PMID- 11427800 TI - [Cutaneous aseptic abscesses, manifestations of neutrophilic diseases]. AB - BACKGROUND: Neutrophilic skin disease includes several entities: Sweet syndrome, pyoderma gangrenosum, erythema elevatum diutium, Sneddon-Wilkinson sub-keratous pustulosis, and neutrophilic eccrine hidradenitis. We report two cases of aseptic abscesses which correspond to the deepest anatomoclinical form of neutrophilic dermatosis. CASE REPORTS: A 28-year-old man was hospitalized for fever and abdominal pain with bloody diarrhea in relation with Crohn's disease. The patient also presented two skin abscesses on the lower limbs. Bacteriology specimens were negative. The histology specimen of a skin lesion revealed neutrophil infiltration of the hypodermis without granulomatosis. Systemic corticosteroid therapy was given and rapidly led to resolution of the inflammatory bowel disease and the skin lesions. The patient developed inflammatory spondylarthropathy several months later. The second patient was a 36-year-old woman with a history of splenomegaly with asceptic abscesses. She was admitted for abdominal pain with non-bloody diarrhea, fever and multiple joint pain related to spondylarthropathy. She developed several simultaneous abscessed nodules on the legs. Biopsy revealed neutrophil infiltration of the hypodermis. The diagnosis of neutrophilic disease with aseptic cutaneous and visceral abscesses was retained. Nonsteroidal antiinflammatory drugs and dapsone were given leading to regression of the skin lesions and the abdominal and joint pain. DISCUSSION: Aseptic skin abscesses result from a deep localization of neutrophilic disease. They suggest the presence of inflammatory bowel disease, spondylarthropathy or other aseptic visceral localizations. PMID- 11427801 TI - [Paraneoplastic pemphigus: a pustular form during chronic lymphoid leukemia]. AB - BACKGROUND: Paraneoplastic pemphigus is an autoimmune disease of the skin and mucosa described in 1990. The condition is generally associated with lymphoma or chronic lymphoid leukemia. Lesions are often misleading, masquerading as polymorphous erythema or lichen. We report a case of paraneoplastic pemphigus with pustulous skin lesions. CASE REPORT: A 52-year-old man developed over a few weeks time erosive lesions of the oral cavity and lips associated with papulous skin lesions. Secondarily, large-sized pustules, sometimes a hypopion, were observed associated with bullae. The diagnosis of paraneoplastic pemphigus was confirmed by direct immunofluorescence that evidenced IgG deposits within the keratinocytes and along the basal membrane and by indirect immunofluorescence on rat bladder that evidenced circulating antibodies. This paraneoplastic pemphigus was the inaugural sign of chronic lymphoid leukemia. DISCUSSION: Skin lesions described in paraneoplastic pemphigus include: erosion, vesicles, bullae, and psoriasiform, lichen-like, plate-like or vegetative formations. To our knowledge, this is the first report of a pustulous form; clinically similar to Hallopeau pustulous pemphigus. PMID- 11427802 TI - [Pigmented Paget's disease of the male nipple]. AB - BACKGROUND: Mammary Paget's disease unfrequently occurs in males, and may be pigmented in rare instances. Differential diagnosis with malignant melanoma relies on immunohistochemical studies. CASE REPORT: A case of Paget's disease of the nipple in a 76 year-old male is reported, clinically mimicking a malignant melanoma because of massive pigmentation. Histologically, large Paget's clear cells were intermingled with numerous melanin-rich dendritic melanocytes. An underlying ductal carcinoma was found. After differential immunohistochemical staining, diagnosis of Paget's disease could be unequivocally substantiated since Paget's cells stained for epithelial markers, c-erbB-2 and hormonal receptors, whereas protein S100 and HMB45 were negative. DISCUSSION: Pigmentation in mammary Paget's disease occurs preferentially in males. Pigmentation results from numerous melanocytes with abundant melanin in close contact with Paget's cells. An increased number of melanocytes may also be observed in cutaneous metastatic breast carcinomas. It could result from a chemotactic factor produced by neoplastic cells. PMID- 11427803 TI - [Necrotizing dermatitis due to Vibrio Vulnificus]. AB - BACKGROUND: Vibrio vulnificus is a Gram-negative halophilic pathogen for man that can cause septicemia in patients with chronic liver disease. In healthy subjects, infections are generally local or regional. We report two cases of necrotizing dermo-hypodermitis due to Vibrio vulnificus. CASE REPORTS: The first patient was a 21-year-old fisherman who developed extensive necrotizing dermo-hypodermitis 24 hours after a penetrating scratch with a fish bone. The second patient was also a fisherman. This 35-year-old man developed a massive area of necrotic tissue on the left leg, also 48 hours after a penetrating fish bone scratch. Both patients exhibited neutrophil hyperleukocytosis and thrombocytopenia. Bacteriological study of the pus and damaged tissue identified Vibrio vulnificus. Blood cultures were negative. Liver tests and liver ultrasound were normal. The course was favorable in both cases with antibiotic treatment and excision of the necrotic tissue. DISCUSSION: For these two cases of Vibrio vulnificus necrotizing dermo hypodermitis, the diagnosis was based on epidemiological, clinical and bacteriological arguments. Neither of these patients had liver disease, explaining the limited locoregional nature of the infection. The severity of the skin necrosis and its rapid course are related to the toxins produced by V. vulnificus. Curative medical and surgical care is generally effective in cases without septicemia and prognosis is good. Education of exposed subjects, fishermen and patients with chronic liver disease, should be helpful for prevention. PMID- 11427804 TI - [Primary infection with West-Nile virus]. AB - BACKGROUND: The recent outbreak of an epidemic of West-Nile fever in New-York revealed this virus' responsibility in some cases of fatal encephalitis. However the clinical manifestations of the infection may be mild as in the case reported herein. CASE-REPORT: A 41-year-old woman without previous medical history presented an acute febrile rash after a trip to Senegal. The course was spontaneously favorable after 5 days, with a biphasic fever at day 3. Reverse passive hemagglutination test confirmed the diagnosis of acute West-Nile fever. DISCUSSION: West-Nile virus, a mosquito-borne flavivirus is usually responsible for a flue-like disease presenting a maculopapular rash in half of the cases. The biphasic evolution of fever is suggestive of the diagnosis. The diagnosis relies on serological tests but cross-reactivities with heterologous flaviviruses may render the interpretation difficult. A rapid recovery is usual but fatal meningoencephalitis can occur especially in the elderly. Treatment is supportive and prevention is limitation of human exposure. West-Nile virus is widely distributed (Africa, Asia, Middle East), but is also endemic in european areas and responsible for seasonal infections. PMID- 11427805 TI - [Iron and Kaposi's sarcoma]. PMID- 11427806 TI - [Hypertrophic lingual papillae]. PMID- 11427807 TI - [A brown plaque on the back]. PMID- 11427808 TI - [Bartonellosis]. PMID- 11427809 TI - [Lymphedema in children]. PMID- 11427810 TI - [Management of bullous pemphigoid]. PMID- 11427811 TI - [The external carotid]. PMID- 11427812 TI - [Basocellular carcinoma and congenital angiomatosis]. PMID- 11427813 TI - [Dissecting cellulitis of the scalp: treatment by isotretinoine]. PMID- 11427814 TI - [Vulvar lichen sclerosus in a little girl]. PMID- 11427815 TI - [Aggravation of asthma by isotretinoine]. PMID- 11427816 TI - [Gynecomastia and finasteride]. PMID- 11427817 TI - [Analysis of an Internet site. Ministry of Health]. PMID- 11427818 TI - [The worldwide challenges of "new" or reemerging communicable diseases at the dawn of the 21st century]. AB - In spite of the very significant advances made during the 20 th century in the prevention and the treatment of communicable diseases, infections are still today, even in developed countries, a major cause of morbidity and mortality. New infectious diseases have emerged (AIDS, legionellosis, exterotoxigenic E. coli, Ebola fever), others have significantly reemerged (tuberculosis, diphtheria, Bartonella infections) or have seen their geographic distribution widen considerably (dengue, Hantavirus, West Nile Virus, Lyme disease). New and widespread hepatotropic viruses (mainly hepatitis C) have been identified, while the bacterial cause (Helicobacter pylori) of gastric ulcer was demonstrated. The second part of this review will deal with other examples of emerging or reemerging infections and with the problem of the increasing resistance of pathogens to antimicrobial agents. It will analyse the multiple causes of these various phenomena and describe the diverse strategies which should become available for the prevention and/or treatment of these numerous infectious diseases. PMID- 11427819 TI - [Mechanism of action and therapeutic uses of botulinum and tetanus neurotoxins]. AB - Botulinum neurotoxins are produced by anaerobic spore forming bacteria, Clostridiumbotulinum. They are synthesized as a single chain protein (150kDa) which is not or weakly active. The active form results from proteolysis that cleaves the precursor into a light chain (about 50kDa) and a heavy chain (about 100kDa) which are linked by a disulfide bridge. The heavy chain is involved in the recognition of a specific neurone surface receptor and mediates the internalization of the light chain into the cytosol. The light chain is responsible for the intracellular activity. It catalyzes the proteolysis of SNARE proteins which are involved in the exocytosis of synaptic vesicles containing acetylcholine. Hence, the release of acetylcholine at the neuromuscular junction is blocked leading to a flaccid paralysis. The tetanus neurotoxin shares with botulinum neurotoxins a common structure and mechanism of action. Tetanus neurotoxin blocks the release of neurotransmitters in the inhibitory interneurons leading to spastic paralysis. The paralytic properties of the botulinum neurotoxins are used to treat certain myoclonies such as blepharospasm, torticolis, hemifacial paralysis. Botulinum neurotoxins are thus efficient therapeutic agents helpful in avoiding surgery. PMID- 11427820 TI - [Inflammatory cascade response to toxin release: therapeutic perspectives]. AB - Release of toxins in the organism trigger a cascade of biological and chemical events. The process involves a large number of molecules produced under genetic control in a perfectly regulated chronological order. Certain molecules (TNFx, IL 1, Il-2.) have inflammatory proprieties, generally producing systemic Inflammatory Response Syndrome (SIRS). Other less numerous molecules (IL-4, IL 10) have antiinflammatory actions. Finally, soluble receptors and these cytokines contribute to the decrease in the quantity of active cytokines at the receptor level. Dozens of other molecules, many of which remain to be fully understood, are also found in the blood stream. They can, for example, facilitate white cell adhesion (ICAM, VCAM, selectins.). Some of them (G-CSF. CM-CSF. IL-5) stimulate production of granulocytes, monocytes, eosinophils. More recently, certain peptides, like macrophage inhibiting factor (MIF) and procalcitonin (PCT) have been added to the list of molecules involved in bacterial infections. Coagulation factors are also very rapidly released in response to toxinic aggression triggering disseminated intravascular coagulation. Later, acute-hase inflammation proteins, such as C-reactive protein (CRP), haptoglobin, serum amyloid A, etc. are found in largely increased quantities. All these molecules are potential markers of inflammation. Only a few have however been retained for routine assay due to their fragility and their short-half life or due to analytical difficulties. CRP and PCT can however be used to differentiate viral infections from bacterial infections and are thus routinely used in clinical applications. In the second part of this review, we briefly discuss therapeutic perspectives for severe infections and septic shock. There have been many attempts to neutralize different cytokines but results have been disappointing to date. There are however many possibilities currently under study, particularly neutralization of endotoxins, immunomodulation, use of recombinant C and S proteins, etc. PMID- 11427821 TI - [Cellular and acellular anti-pertussis vaccines]. AB - Since 1940, vaccination with whole cell vaccines, composed of Bordetella pertussis extracts, has dramatically reduced the incidence of whooping cough. However, the occurrence of side effects has stimulated research that has resulted in the design of new, acellular vaccines. These vaccines are composed of adhesins, such as filamentous haemagglutinin and pertactin, and detoxified pertussis toxin. Detoxification can be achieved by genetic engineering resulting in alterations of specific amino acid residues that are involved in the enzymatic activity or in the target-cell receptor binding activity of the toxin. Assessment on laboratory animals and in clinical trials have indicated that these acellular vaccines are almost as protective as the whole cell vaccines, and that they are much less reactogenic. However, these vaccines are more expensive and still need multiple injections to achieve optimal protection. An interesting alternative would be the construction of attenuated strains able to protect after a single intranasal administration. In addition, the construction of recombinant attenuated strains would perhaps simultaneously protect against whooping cough and other infectious or parasitic diseases. PMID- 11427822 TI - [Synthesis and pharmacological study of 7-phenyl-1,4-diazepin-5-one and its derivatives]. AB - We studied the synthesis and psychotropic activity of the 7-phenyl-1,4-diazepin-5 one and derivatives. It can be conclude that these products have sedative, myorelaxant and anxiolytic actions. The toxicity study demonstrated that two diazepines are non-toxic at therapeutic dosages but that a third compound is very toxic. PMID- 11427823 TI - [Antihypertensive effect of Herniaria glabra saponins in the spontaneously hypertensive rat]. AB - We studied the antihypertension action of Herniaria glabra saponins and compared the effect with that of furosemide. Spontaneously hypertensive rats were treated with H. glabra saponins at a dosage of 200mg/Kg of body weight. Treatment led to progressive decline in both systolic and diastolic blood pressures. After one month of therapy, pressures were: 141,60+/-7,51 / 90,40+/-5.46mmHg versus 187,60+/-5,94/119,10+/-7.79mmHg (p<0,001). We did not observe any change in heart rate. H. glabra saponins were able to lower blood pressure by a multifactorial mechanism. PMID- 11427824 TI - [Elaboration of community healthcare services: an opportunity to prospectively reflect on the healthcare system]. PMID- 11427825 TI - [Impaired fertility in women infected with HIV-1. Implications for sentinel serosurveillance]. AB - BACKGROUND: Given the relationship between HIV infection and fertility, antenatal clinic-based HIV prevalence may not provide a good estimate of the community HIV prevalence. The objective of this work was to evaluate the impact of HIV infection on fertility among women attending antenatal clinics in Bobo-Dioulasso (Burkina Faso), and to discuss possible implications on HIV sentinel surveillance. METHODS: In the context of a phase II/III clinical trial of a short course of Zidovudine during pregnancy (DITRAME - ANRS 049 trial) we consecutively proposed voluntary counselling and HIV testing (VCT) to 1349 women aged at least 18 years, carrying a pregnancy of 7 months or less and living in Bobo-Dioulasso. During pre-test counselling session, a standardised questionnaire was administered to collect detailed information regarding socio-demographic characteristics and obstetrical history. Blood samples were then taken and tested for HIV after written informed consent. RESULTS: Mean age (+/- standard deviation) at first sexual intercourse was similar among HIV-infected (HIV+) (16.7+/- 2; n=83) and HIV-negative (HIV-) women (16.9+/- 2; n=1336). However, HIV+ women aged 25 years and above had, on the average fewer pregnancies (3.8+/- 1.5; n=37) than HIV- women (5.0+/- 2.3; n=567), p<0.01. Similarly, these HIV+ women had, on average, less live births (2.8+/- 1.3; n=35) than HIV- ones (3.7+/- 2.1; n=555), p=0.02. Other sexual and obstetrical characteristics such as maternal age, proportion of primigravidae, stillbirths or spontaneous abortions were comparable between HIV+ and HIV- women. CONCLUSIONS: Our data suggest that the level of fertility of HIV+ women aged 25 years and above is significantly lower than for HIV- women. Therefore, HIV+ women in this age group are likely to be under-represented among antenatal clinic attendees. These findings suggest adjusting antenatal clinic-based HIV sentinel surveillance data for age and fertility in order to derive a good estimate of the community HIV prevalence. PMID- 11427826 TI - Social and health status of arrivals in a French prison: a consecutive case study from 1989 to 1995. AB - BACKGROUND: To assess the demographic, socioeconomic and health status of male arrivals in French jails and to analyze the time trends of these characteristics. METHODS: The study was carried out in a prison for detained persons and short term prisoners. Using a standardized questionnaire, we recorded the characteristics of all male detainees and prisoners arriving in the prison between 1989 and 1995. The information collected concerned: demographic data, level of education and professional status, reasons for detention or imprisonment, social and family background, lifestyle, medical and psychiatric history, suicide attempts and illicit use of drugs. The characteristics of the study population were compared with those found in the general regional population. We analyzed developing trends in the health status of the prison population as well as their socio-economic profile over a period of seven years (1989 to 1995). RESULTS: A total of 14,785 questionnaires were analyzed. Of the study population, 56% had no professional qualification, and 62% was unemployed. About two-thirds of the inmates presented psychiatric problems or problems with illicit drug use (one or several drugs). Amongst these, 70% had not had any form of care -neither therapeutic nor educational- before their arrival in prison. Between 1989 and 1995, the proportion of drug users increased by 30%. A parallel increase was observed in the level of unemployment and in the frequency of mental problems. CONCLUSIONS: Our results suggest a need for therapeutic and educational care to be provided for prison inmates. This poses a problem which needs to be addressed in terms of public health. The study also illustrates the usefulness of a standardised questionnaire for each arrival. The systematic use of such a tool would make it possible to identify inmates'needs and to propose adapted care solutions. PMID- 11427827 TI - [Declared knowledge, beliefs and practices for cardiovascular disease prevention in the French population]. PMID- 11427828 TI - [Cost-benefit ratio of tick-borne encephalitis vaccination of French troops based in Kosovo]. AB - BACKGROUND: French military troops based in Kosovo are exposed to Tick-Borne Encephalitis (TBE) biorisk. An efficacious vaccine is available to prevent this disease, burdened by high morbidity and mortality. The epidemiologic features of TBE in Kosovo are unprecise. To help the French Department of Defense to make a decision about immunization against TBE, we performed a cost-benefit analysis. METHODS: By a Bayes'analysis decision, we have estimated the net benefits for a three-injection vaccine program versus absence of vaccination, for all French military troops based in Kosovo. Time expectancy was 4 years. Through the review of several medical reports, we have estimated the useful parameters for this study: morbidity incidence rate, mortality rate, sequelae rate, efficacy vaccine and side effects. We have chosen as initial hypothesis a sero-conversion rate of 1,080 per 100,000 men-by-year. Human life was valorized in francs, by calculating the allowance paid by Department of Defense to the family in case of death or for sequelae. RESULTS: Net benefits arised to 2.17 millions of francs. The vaccine program cost was 25.0 millions of francs. 140 cases of encephalitis viruses were saved by vaccination. Nevertheless a sensibility analysis has shown that results are well reliable with TBE incidence rate. CONCLUSION: The break even point being close to the incidence rate of the initial assumption, the decision to vaccine all French military troops depends at the same time on a better knowledge of the incidence of the disease in Kosovo, but also of the number of potentially avoided cases of tick-borne encephalitis and of the programs of vaccination concerning this disease of the other countries forming the United Nations Organization forces. PMID- 11427829 TI - [Medical and economic evaluation of organ transplantation in France. The example of liver transplantation]. AB - BACKGROUND: Organ transplantation is an expensive and risky medical procedure. Estimating the cost is difficult because the care is complex and involves many actors. We present a methodological framework for the economic evaluation of organ transplantation in France which include a detailed collection of all the direct costs and the simultaneous analysis of health status. It was applied to evaluate the cost of liver transplantation in France. METHODS: All consecutive adults transplanted or placed on the waiting list of liver transplantation in the Department of Surgery of Cochin Hospital, Paris, between 1994 and 1996 were included. All resource use was measured during one year: staff wages, pharmacy and blood, laboratory and radiology, supplies, overhead hospital services. Mean quality of life was estimated by the survival weighted by the Karnofsky index. RESULTS: Transplantation: 38 patients were included. The first year mean cost after transplantation was 561,000FF (included rehospitalizations cost of 120,000FF). Care outside the hospital induced 10% of the total cost. Mean quality of life was 63% (from 0% to 93%) and increased with time, whereas cost decreased. Waiting list: 26 of 33 patients on waiting list were transplanted. The first year mean cost was 95,000FF (included 32,000FF for first evaluation) and increased with time. 44% of cost was supported by another hospital than the transplanting one. Sickness allowance added 20% to the cost. The mean quality of life was 56% during the first year. CONCLUSIONS: This complete approach of organ transplantation cost respected the medical procedure over the time. Detailed costs take into account the care outside the hospital. This method can be used in other countries and generalised to all surgical or medical procedure as heavy as the organ transplantation. PMID- 11427830 TI - A method for shortening instruments using the Rasch model. Validation on a hand functional measure. AB - BACKGROUND: Many measurement instruments, particularly measures of hand functional ability, frequently comprise a large number of items. Reduced versions of these instruments can facilitate their use. This work proposes a new method for shortening an instrument. METHODS: The method proposed was based on a scale of item difficulty calculated using the Rasch model. It was applied on a hand functional measure comprising 67 tests. The sample included 194 patients with hand lesions. The shortened instrument obtained was compared with those provided by classic methods used in the literature, with item random choice, and with shortened versions proposed by four independent experts, two rehabilitation physicians and two occupational therapists, who are clinicians familiar with the tool. All the statistical analyses were carried out on a random sub-group of two thirds of the sample. A cross validation was then carried out on the remaining third. RESULTS: The reduction obtained had score non significantly different from that of the original instrument. In addition, the intra-class correlation coefficient and the Cronbach alpha coefficient were high. Among the different degrees of reduction investigated, the 12-item version seemed to be appropriate. Our method appeared to provide better results in terms of discriminant validity and internal validity than the choices of the four experts. The reductions produced were also better than those obtained by classic methods based on principal component analysis and multiple linear regression, as well as those obtained by random choices of items. CONCLUSION: The method presented is pertinent and useful. The reduction obtained appeared to be better than the choices of experts and the reductions provided by classic methods. The method could be used in other fields. PMID- 11427831 TI - [Comorbidity indexes: review of the literature and application to studies of elderly population]. AB - BACKGROUND: Older patients often suffer concomitant chronic diseases in addition to various health disorders. When attempting to study one of these diseases, it is necessary to consider the complexity of the patient's health status. Comorbidity is a cause of confusion, especially for elderly patients with multiple diseases. Comorbidity indexes have been developed in order to measure this complexity. METHODS: We focused this article on a review of the literature in order to identify and examine the validity of comorbidity indexes applied to the elderly population. RESULTS: Five indexes have been used to measure comorbidity in patient populations with complex health situations: the Cumulative Illness Rating Scale (CIRS), the Kaplan-Feinstein index, the Charlson index, the Index of Coexistent Disease (ICED), and the Total Illness Burden Index (TIBI). Each of these indexes included a series of domains which vary according to the authors'view of comorbidity. These indexes were validated under different conditions with similar targets. Their validation fields limited their use and extrapolation of results. Only the CIRS, the Charlson index, the Kaplan-Feinstein index and the ICED were validated and applied to old patients. However, the Charlson index was found to be limited in recording the entirety of the old patients'pathologies, and in patients with cognitive deficits, only CIRS appeared to be sufficiently trustworthy because it allows a comprehensive recording of all the comorbid disease from clinical examination and medical file data. CONCLUSION: When studying chronic disease states in frail elderly patients, it is essential to consider comorbidity using standard validated indexes in order to get a comprehensive assessment of the patient's situation and avoid neglecting diseases and handicaps. Further studies are necessary to develop and validate tools specifically adapted to the elderly population. PMID- 11427832 TI - [How to evaluate the quality of information transfer from physician to patient? Choice of psychometric tests for a decision tree in a Regional Cancer Centre]. AB - BACKGROUND: Decision boards are used to transfer information from physicians to patients to help them participate in the clinical decision-making process. We present the tests and results of the psychometric properties of a decision board in a sample of healthy volunteers. METHODS: In the Regional Cancer Centre located in Lyon, we developed a decision board for post-menopausal women with breast cancer after lumpectomy without any poor prognostic factors. Two treatment options were proposed, one involving chemotherapy and the other not. We tested for the following psychometric properties: comprehension, construct validity and reliability. Comprehension was evaluated using a questionnaire, in order to test whether the rates of correct answers were due to chance alone. The construct validity was assessed by changing the information provided (relapse and survival rates, characteristics of chemotherapy) and testing whether the proportion of healthy volunteers choosing an option changed in a predictable and significant way. The reliability was evaluated using the test-retest method. Two reliability statistics were computed: the Pearson correlation and the Intraclass Correlation Coefficient. RESULTS: In the sample of 40 healthy volunteers, 23 chose the option with chemotherapy and 17 the option without chemotherapy. Results show that the decision board was comprehensive, valid (the women changed their choices in a predictable way) and reliable (Pearson correlation and Intraclass Correlation Coefficient close to 1). CONCLUSION: The choice of the psychometrics properties tested and the statistical tests used are discussed. The psychometric properties of our tool are found to be satisfactory. PMID- 11427833 TI - Safer colonoscopy with patient-controlled analgesia and sedation with propofol and alfentanil. AB - BACKGROUND: The aim of this study was to assess the efficacy of patient controlled analgesia and sedation with propofol/alfentanil for colonoscopy compared with continuous drug infusion and conventional nurse-administered medication. METHODS: One hundred fifty patients undergoing colonoscopy on an outpatient basis were randomly assigned to 1 of 3 medication regimens. To maintain blinding, all patients were connected to an infusion pump. Group I patients could self-administer boluses of 4.8 mg propofol and 125 microg alfentanil without restriction. Group II patients received a continuous infusion with 0.048 mg/kg propofol and 0.12 microg/kg alfentanil per minute. Group III patients received intravenous premedication with 0.035 mg/kg midazolam and 0.35 mg/kg meperidine. RESULTS: There were no differences between the groups with respect to pain (visual analogue scale) and procedure time. Patient-controlled analgesia and sedation with propofol/alfentanil (group I) resulted in less of an increase in the transcutaneous partial pressure of carbon dioxide (p = 0.0004) during colonoscopy and less of a decrease in mean arterial blood pressure (p = 0.0021) during recovery, as well as more complete recovery (p = 0.0019) after 45 minutes compared with conventional administration of midazolam/meperidine. Furthermore, patient-controlled analgesia and sedation yielded a higher degree of patient satisfaction than continuous infusion of propofol/alfentanil (p = 0.0033) or nurse-administered midazolam/meperidine (p = 0.0094). CONCLUSIONS: Patient controlled administration of propofol and alfentanil for colonoscopy may provide a better margin of safety than conventional administration of midazolam and meperidine and results in a higher level of patient satisfaction and shorter recovery. PMID- 11427834 TI - Patient-controlled sedation with propofol for colonoscopy. AB - BACKGROUND: Patient-controlled sedation (PCS) allows the patient to titrate the dosage of sedative drugs according to need. METHODS: To compare the efficacy of PCS by using propofol with anesthetist-administered midazolam during colonoscopy, 88 patients were randomized to receive either intravenous midazolam 0.05 mg/kg bolus (1 mg increments as required) or propofol PCS (0.3 mg/kg bolus, zero lockout). Heart and respiratory rates, blood pressure, and oxygen saturation were monitored. Patient cooperation, endoscopist satisfaction, and level of sedation were scored. Patient satisfaction was assessed by questionnaire. To correct for multiple testing of data, statistical significance was asserted only for individually stated p values with p < 0.01. RESULTS: Oxygen saturation and hemodynamics were stable in both groups. Better patient cooperation (good vs. minimal; p = 0.008) and higher endoscopist satisfaction (very good vs. good; p = 0.001) were achieved with PCS. Although more sedated intraoperatively (sedation score 4 vs. 2; p = 0.03 for a single test of hypothesis; correction for multiple testing of data removes this significance), patients in the propofol PCS group were more alert by 30 minutes and discharged earlier (mean and SD times were 43.3 [12.1] min compared with 61.0 [29.7] min; p = 0.001.) More patients in the PCS group (86 vs. 6l%; p < 0.001) were satisfied with their overall level of comfort. CONCLUSION: PCS with propofol is effective and results in high patient satisfaction and faster discharge. PMID- 11427835 TI - Patient-maintained sedation for ERCP with a target-controlled infusion of propofol: a pilot study. AB - BACKGROUND: ERCP requires patient cooperation and often prolonged sedation. In different areas of anesthetic practice, patient-controlled sedation with a target controlled infusion (TCI) of propofol provides effective sedation. The aim of this study was to assess the safety and efficacy of the same system in patients undergoing ERCP. METHODS: Twenty patients used the TCI system. Patients received oxygen at 2 L/min via nasal cannulae. By using pharmacokinetic TCI software modeling, an initial propofol target blood concentration (Ct) of 1.0 microg/mL was supplemented on patient demand with a handset that, when pressed twice within 1 second, increased the Ct of propofol by 0.2 microg/mL. The maximum permissible target concentration was set at 3.0 microg/mL to prevent oversedation. RESULTS: Sixteen patients used the system successfully throughout the procedure. The Ct propofol ranged from 1.2 to 2.6 microg/mL, and the number of successful handset activations (after commencement of the ERCP) ranged from 0 to 3. In 3 patients, the ceiling Ct propofol was attained without adequate sedation and the system was manually overridden. The system failed in 1 case because of patient confusion. There were no episodes of hemodynamic instability, airway obstruction, or significant oxygen desaturation. Endoscopist and patient satisfaction were high. Four patients were oversedated according to our criteria at the end of the procedure, but all were awake within 5 minutes of arrival in the recovery area. CONCLUSIONS: Patient-maintained sedation with TCI propofol was safe and fully effective in 16 patients. Ease of endoscopy was rated high by the endoscopists, and all patients were well satisfied with their sedation. Adjustments to the software programming are being evaluated to increase the safety profile to avoid oversedation. PMID- 11427836 TI - Feasibility and potential benefit of maintenance endoscopic variceal ligation in patients with unresectable hepatocellular carcinoma and acute esophageal variceal hemorrhage: a controlled trial. AB - BACKGROUND: Patients with unresectable hepatoma and acute esophageal variceal bleeding have extremely high rates of recurrent bleeding and mortality. This controlled study evaluates the feasibility and potential benefit of maintenance endoscopic variceal ligation in these patients. METHODS: Patients with unresectable hepatoma and acute esophageal variceal bleeding underwent emergent endoscopic variceal ligation. After hemostasis, patients were randomized to undergo maintenance or esophageal variceal ligation (EVL) as necessary (demand ligation). RESULTS: Fifty-four patients underwent maintenance EVL and 55 demanded EVL. One or more subsequent EVL session could be performed in only 30 patients (55.6%) in the maintenance group (actual maintenance ligation). Logistic regression analysis found that hepatic function determines the feasibility of maintenance ligation (Child-Pugh's A+B vs. C, OR 23.00: 95% CI [5.26, 100.66]). The survival and recurrent bleeding rates were similar in both groups. A subgroup analysis of patients with Child-Pugh's A and B hepatic reserve in both the maintenance EVL group (n = 24) and demand EVL group (n = 25) was performed to assess the potential benefit of maintenance ligation. Maintenance ligation reduced the rate of recurrent bleeding compared with demand ligation (p = 0.043). Cox regression showed that portal vein thrombosis and tumors in both hepatic lobes were also factors together with EVL that determined recurrence of bleeding. Survival was similar in both groups. CONCLUSIONS: Maintenance ligation is feasible in patients with unresectable hepatoma and variceal hemorrhage if they have a good hepatic reserve. Maintenance ligation might lower the rate of recurrent bleeding in this subgroup of patients. PMID- 11427837 TI - Validation of color Doppler EUS for azygos blood flow measurement in patients with cirrhosis: application to the acute hemodynamic effects of somatostatin, octreotide, or placebo. AB - BACKGROUND: Color Doppler EUS (CD-EUS) allows minimally invasive measurement of azygos blood flow (AzBF) in portal hypertension, but further validation of the method is needed. Because a limited number of patients has been studied, the acute hemodynamic effects of somatostatin and octreotide on AzBF and gastric mucosal perfusion are poorly defined in portal hypertension. METHODS: A double blind hemodynamic study was designed to assess rapid changes in AzBF over a 60 minute period after intravenous administration of somatostatin, octreotide, and placebo in 30 stable patients with biopsy-proven cirrhosis. AzBF was measured by using both CD-EUS and the invasive thermal dilution technique in the first 10 patients (phase 1). Then, with CD-EUS alone, the hemodynamic study was extended to a further 20 patients (phase 2). In addition, gastric mucosal perfusion changes were assessed by using laser Doppler flowmetry at endoscopy. RESULTS: In phase 1, the 2 methods for AzBF measurement showed significant correlations both for baseline values (r = 0.685) and for AzBF changes over 60 minutes after drug administration (r = 0.733). In phase 2, a reduction was observed in AzBF 10 minutes after octreotide or somatostatin administration (-47% and -23%, p < 0.0001 vs. placebo, p = 0.058 vs. placebo, respectively). After 60 minutes of somatostatin infusion, AzBF increased 27% over placebo values (p < 0.04). Gastric mucosal perfusion was transiently reduced 5 minutes after octreotide or somatostatin (-21% and -32%, respectively, p < 0.02 vs. placebo). CONCLUSIONS: This is the first study to validate CD-EUS AzBF measurement with reference to the invasive thermodilution technique in cirrhosis. It confirmed the transient effects of somatostatin and octreotide on both AzBF and gastric mucosal perfusion. In addition, a significant rebound phenomenon after 60 minutes of continuous intravenous somatostatin infusion was observed. PMID- 11427838 TI - Wire-guided intraductal US: an adjunct to ERCP in the management of bile duct stones. AB - BACKGROUND: Endoscopic retrograde cholangiography (ERC) may misdiagnose bile duct stones if air bubbles are introduced during contrast injection, and it may also fail to diagnose stones in the presence of bile duct dilation. METHODS: Our aim was to determine whether intraductal US (IDUS) improves the accuracy of cholangiography and whether it is a useful adjunct in the management of bile duct stones. IDUS with a wire-guided US probe was performed after initial ERC in patients in whom bile duct stones were suspected. The diagnostic accuracy of ERC alone was compared with that of ERC plus IDUS. RESULTS: ERC with IDUS was performed in 62 patients who were suspected to have bile duct stones. Both IDUS and ERC were performed by the same endoscopist, and ERC was performed with a C arm fluoroscope. The presence of bile duct stones and/or sludge were confirmed after sphincterotomy and extraction in 34 patients. Overall, the accuracy of ERC combined with IDUS in the diagnosis of bile duct stone and/or sludge was higher than that of ERC alone (97% vs. 87%, p < 0.05). With dilated bile ducts, the diagnostic accuracy of ERC combined with IDUS was also higher than that of ERC alone (95.5% vs. 72.7%, p < 0.05). Additional diagnostic information provided by IDUS included identification of cystic duct stones in 5 patients, characterization of bile duct strictures in 2 patients, and choledochal varices in 1 patient. Performance of wire-guided IDUS required 5% of the total procedure time. CONCLUSIONS: IDUS improves diagnostic accuracy of ERC and is a useful adjunct to ERC when bile duct stones are suspected. PMID- 11427839 TI - The frequency of bile duct crystals in patients with presumed biliary pancreatitis. AB - BACKGROUND: Previous studies of biliary microlithiasis in acute pancreatitis of uncertain etiology were conducted a few weeks to months after the acute episode. Bile obtained during urgent ERCP (less than 24 hours after admission) was studied for the presence of microlithiasis during the acute phase of acute pancreatitis of suspected biliary origin. METHODS: Fifteen consecutive patients with acute pancreatitis of suspected biliary origin were recruited from a population of 309 patients with acute pancreatitis (5%) treated during the last 4 years. Patients with gallstones on US and/or ERCP and those in whom the etiology of acute pancreatitis was certain were excluded. RESULTS: Microlithiasis (mostly calcium bilirubinate granules) was found in 12 (80%) cases. Despite endoscopic sphincterotomy 3 patients died within 2 weeks because of multisystemic organ failure. Among the 12 remaining patients, 2 (16%) developed gallbladder stones and 1 underwent cholecystectomy for cholecystitis (8%) during follow-up. The average length of follow-up was 30 months. No episodes of acute pancreatitis were noted during follow-up. CONCLUSIONS: In the acute phase of acute pancreatitis of suspected biliary origin, biliary microlithiasis was found in most cases. Endoscopic sphincterotomy appears to protect patients from further episodes of acute pancreatitis. PMID- 11427840 TI - Risk factors for recurrence of primary bile duct stones after endoscopic biliary sphincterotomy. AB - BACKGROUND: Endoscopic biliary sphincterotomy (EST) is a well-established procedure for bile duct stone extraction. Bile duct stones can be classified as primary or secondary. However, few data are available on the recurrence of primary and secondary bile duct stones after EST. Therefore risk factors for the recurrence of primary bile duct stones after EST were prospectively studied. METHODS: Between 1991 and 1997, 61 patients underwent EST for primary bile duct stones. All met the following criteria: (1) previous cholecystectomy without bile duct exploration, (2) detection of bile duct stones at least 2 years after initial cholecystectomy. Mean follow-up was 2.2 years. Fourteen patients were lost to follow-up. The recurrence of primary bile duct stones was defined as the detection of bile duct stones no sooner than 6 months after complete clearance of primary bile duct stones. RESULTS: The overall recurrence rate of primary bile duct stones was 21% (10 of 47). Two significant risk factors for recurrence were identified by multivariate analysis: (1) patients with a bile duct diameter of 13 mm or greater after stone removal had recurrences more frequently than those with a duct diameter of 13 mm or less, and (2) patients whose papilla was located on the inner rim or deep within a diverticulum, so that the papillary orifice was not visible endoscopically, had more frequent recurrences than patients with a papilla outside the diverticulum, or no peripapillary diverticulum. CONCLUSION: The independent risk factors for recurrence of primary bile duct stones were sustained dilation of the bile duct even after complete removal of stones and location of the papilla on the inner rim or deep within a diverticulum. PMID- 11427841 TI - The role of endoscopic retrograde pancreatography in the treatment of traumatic pancreatic duct injury. AB - BACKGROUND: The status of the main pancreatic duct (MPD) is the most important determinant of the morbidity and mortality associated with pancreatic trauma. Early diagnosis and optimal treatment are critical, especially when there is MPD injury. METHODS: Twenty-three patients with pancreatic trauma were studied prospectively with respect to clinical and laboratory findings, CT, and endoscopic retrograde pancreatography (ERP). Treatment modalities and clinical outcome were assessed in relation to ERP findings. RESULTS: The pancreatic duct was injured in 14 of 23 patients (11 MPD, 3 branch duct). Contrast leakage from the MPD into peritoneal cavity at ERP confirmed MPD injury in 8 patients, who underwent surgical exploration. Three patients with leakage from a branch duct into the pancreatic parenchyma recovered with conservative treatment. Three patients in whom ERP demonstrated contrast leakage from the MPD confined to the parenchyma underwent successful transpapillary stent insertion with complete resolution of the leak at 3-month follow-up. Patients who underwent ERP more than 72 hours after trauma had a significantly higher rate of pancreas-associated complications and a tendency to remain hospitalized longer than patients who underwent ERP earlier. CONCLUSION: Early ERP is one of the most useful methods for demonstrating MPD injury. ERP assists with treatment planning based on the degree of pancreatic duct injury. PMID- 11427842 TI - The relationship between juxtapapillary duodenal diverticula and disorders of the biliopancreatic system: analysis of 350 patients. AB - BACKGROUND: Data concerning the association of juxtapapillary duodenal diverticula (JPDD) with biliopancreatic disorders are inconsistent, but an association between bile duct stones and JPDD is widely accepted. The aim of this study was to investigate the frequency of JPDD and its association with biliopancreatic disorders in patients undergoing ERCP. METHODS: A retrospective analysis was conducted of 5497 consecutive ERCP procedures performed in 2925 patients. Matched-pair analysis yielded 350 pairs of patients with and without JPDD, matched for definite risk criteria such as age, gender, and indication for ERCP. RESULT: The incidence of JPDD was 12%. Patients with JPDD were significantly older than patients without JPDD (71 vs. 62 years; p < 0.0019) and had a significantly higher bleeding rate after endoscopic sphincterotomy (8.8% vs. 4.8%; p = 0.039). The presence of JPDD correlated with gallbladder stones (29.4% vs. 20.8%; p = 0.039), bile duct stones (46% vs. 33.1%; p < 0.001), and recurrence of bile duct stones (6.6% vs. 1.4%; p = 0.002). There were no significant differences in frequency of acute and chronic pancreatitis as well as pancreas divisum. After multivariate logistic regression analysis, technically difficult ERCP, bleeding after endoscopic sphincterotomy, and bile duct stones remained as independent risk factors. CONCLUSION: JPDD appears to be a risk factor for complications of endoscopic sphincterotomy and for gallbladder stones, bile duct stones, and their recurrence. PMID- 11427843 TI - Clinicopathologic features and endoscopic treatment of superficially spreading colorectal neoplasms larger than 20 mm. AB - BACKGROUND: A colorectal neoplasm that spreads superficially over the mucosa is known as a laterally spreading tumor. The clinicopathologic features of these large lesions and the efficacy and safety of endoscopic mucosal resection (EMR) were studied retrospectively. METHODS: Surgically or endoscopically resected laterally spreading tumors larger than 20 mm in diameter were studied. Lesions were divided into 2 macroscopic subtypes: F-type, composed of superficially spreading lesions with a flat and smooth surface, and G-type, composed of superficially spreading aggregates of nodules that form relatively flat, broad based lesions with granulonodular and uneven surfaces. RESULTS: Thirty-three lesions were of the F-type and 87 the G-type. G-type (mean +/- SD, 35.3 +/- 11.4 mm) lesions were significantly larger (p < 0.01) than F-type (26.0 +/- 7.2 mm) lesions. F-type lesions had a significantly higher frequency of invasive cancer (27.2%) than G-types (10.3%)(p < 0.05). Of the 120 lesions, 81 (67.5%) were resected endoscopically. Patients with 78 of these lesions were followed postoperatively for 60.8 +/- 20.1 months. The rate of local recurrence of endoscopically treated tumors as determined at colonoscopy was 7.4% (6/78). These lesions were completely resected endoscopically. Distant metastases were not detected. Thirteen (16.0%) patients had local bleeding after EMR that was stopped endoscopically. Microperforation of the colonic wall as a result of EMR was diagnosed in 1 (1.2%) of 81 cases. CONCLUSIONS: Laterally spreading tumors larger than 20 mm, especially those of the G-type, have a low rate of invasion despite their relatively large size. The F-type lesion has a higher malignant potential than the G-type. EMR is an effective and safe treatment for the large laterally spreading tumor. PMID- 11427844 TI - Comparison of patients' experiences during imaging tests of the colon. AB - BACKGROUND: Physical discomfort, anxiety, embarrassment, and other aspects of patient experience impact on future compliance for colonic imaging tests. Therefore, a prospective study was performed comparing patient experiences during air contrast barium enema (ACBE), flexible sigmoidoscopy, and colonoscopy. METHODS: Immediately after each procedure, patients completed a questionnaire assessing pretest anxiety, difficulty with preparation, pain, cramping, bloating, overall discomfort, loss of dignity, willingness to repeat the test, and overall satisfaction. A follow-up questionnaire was administered within 48 hours. Nurses and physicians also completed questionnaires to assess the provider impression of patient experience. RESULTS: Four hundred ten patients (80 ACBE, 202 sigmoidoscopy, 128 colonoscopy) were prospectively enrolled. Sigmoidoscopy caused more pain than ACBE (Odds ratio [OR] 2.64: 95% CI [1.63, 4.27]) or colonoscopy (OR 1.83: 95% CI [1.21, 2.77]). ACBE and colonoscopy did not differ in the degree of pain. Although overall satisfaction appeared to be similar for all tests, patients were less willing to repeat ACBE than sigmoidoscopy (OR 1.85: 95% CI [1.13, 3.02]) or colonoscopy (OR 1.82: 95% CI [1.07, 3.09]). Initial and follow up pain ratings by patients were highly correlated (Spearman correlation 0.81); however, correlation of pain assessments between staff and patients was poor (Spearman correlation 0.48). CONCLUSIONS: Sigmoidoscopy was more painful than other colonic imaging modalities. Although ACBE and colonoscopy caused similar pain, patients were less willing to repeat ACBE. In aggregate, the data suggest that patients perceive colonoscopy as the most acceptable colonic imaging procedure. Better methods are required to allow staff to adequately assess discomfort experienced by patients during these procedures. PMID- 11427845 TI - Metastatic osteosarcoma involving the colon and ileum. PMID- 11427846 TI - Isolated metastatic gastric cancer to the colon: a unique endoscopic appearance. PMID- 11427847 TI - A case of chronic lymphocytic leukemia-complicated colonic intussusception. PMID- 11427848 TI - Peri-minor-ampullary diverticulum. PMID- 11427849 TI - Electrical stimulation for propelling endoscopes. AB - BACKGROUND: Methods are needed for propulsion of endoscopes and wireless video capsules along the small intestine. This work aims to test the hypothesis that electrical stimulation could propel an endoscope by stimulating muscular contraction. METHODS: Prototype acrylic ovoid-shaped devices were constructed with 2 stainless steel electrodes mounted on the tapered section. Five devices 15 to 23 mm diameter with a taper of 28 degrees to 40 degrees (included angle) were tested. When these devices were in contact with the bowel wall, electrostimulation was applied causing circular muscle contraction, which when applied to the taper of the ovoid resulted in forward propulsion of the device. The method does not induce peristalsis but works by stimulating local contraction. The device was tested in the small intestine and esophagus of anesthetized pigs. RESULTS: Electrostimulation caused the ovoid to advance rapidly (6 mm/sec) up and down the esophagus by inducing circular esophageal muscle contraction. When stimulated at 15 Hz with 30-ms pulses, the threshold for movement in the small intestine was 12 mA; at 20 mA the device moved reliably in either direction in the small intestine at speeds of up to 4.5 mm/sec and negotiated tight curves. CONCLUSION: Electrostimulation can move endoscopes in the small intestine. PMID- 11427850 TI - Wire-assisted minor papilla precut papillotomy. AB - BACKGROUND: Deep cannulation of the dorsal duct in patients with pancreas divisum may not be possible, even for the experienced endoscopist. This is a description of a technique to facilitate access to the dorsal duct. METHODS: Patients undergoing dorsal ductography in whom endoscopic therapy was indicated and in whom a 4.5F tapered-tip catheter could not be passed into the dorsal duct over a guidewire were prospectively identified. After ductal opacification, either a 0.035- or 0.021-inch guidewire was passed into the dorsal duct; a needle-knife was then passed to the minor papilla alongside the guidewire, and with appropriate orientation, a precut papillotomy was made. RESULTS: Eleven patients undergoing dorsal ductography for recurrent pancreatitis (10 patients) or ductal leak (1 patient) were prospectively treated with the above technique. Secretin was used in 1 patient. In all patients, after precut papillotomy the diagnostic catheter was passed without difficulty into the dorsal duct allowing for guidewire exchange and/or stent placement. Extension of the papillotomy could then be performed with the needle-knife over a pancreatic stent. Complications included pancreatitis in 1 patient. CONCLUSIONS: This technique, which uses widely available accessories, facilitates access to the dorsal pancreatic duct for endoscopic therapy. PMID- 11427851 TI - EUS-guided one-step drainage of pancreatic pseudocysts: experience in 3 patients. AB - BACKGROUND: EUS-guided drainage has been recognized as a nonsurgical treatment for pancreatic pseudocysts. Bleeding at the puncture site is a known complication of this procedure. This is a report of the use of new equipment for one-step drainage without the use of an electrosurgical needle. METHODS: EUS-guided cystoenterostomy was performed in 3 patients. The mean size of the pseudocysts was 8 cm (head = 1, body = 2). A needle and drainage tube were inserted in a one step procedure into the cyst through the accessory channel of an echoendoscope under EUS. The drainage tube was released immediately after insertion. OBSERVATIONS: All 3 patients were treated successfully without complication. A drainage tube was inserted through the duodenal bulb (n = l) and the body of the stomach (n = 2). The drain was removed after 1 month in all patients. Two of the 3 drains were obstructed by sludge. One pseudocyst recurrence was observed at a mean follow-up of 3.6 months (range 2-6 months). CONCLUSIONS: The EUS-guided one step system is simple to use and suitable for drainage of pancreatic pseudocysts. It appears to minimize the risk of hemorrhage. PMID- 11427852 TI - Dilation of high-grade pancreatic and biliary ductal strictures with small caliber angioplasty balloons. AB - BACKGROUND: Pancreatic and bile duct strictures may be too stenotic to allow passage of conventional endoscopic dilators. METHODS: Four patients with strictures (3 pancreatic, 1 biliary) that could not be traversed with conventional endoscopic dilating devices, or in 1 case by a Soehendra stent extractor, underwent stricture dilation with a 3.3F peripheral angioplasty balloon to a maximum diameter of 6 mm. OBSERVATIONS: All strictures in the 4 patients were successfully traversed and dilated and stents were placed with resolution of the presenting clinical problem. CONCLUSIONS: Small-caliber angioplasty balloons are useful for dilation with subsequent stent placement of pancreatic and biliary strictures that are refractory to standard endoscopic approaches. PMID- 11427853 TI - Regression of primary lymphoma of the ampulla of Vater after eradication of Helicobacter pylori. PMID- 11427854 TI - Dissolution of an esophageal bezoar with pancreatic enzyme extract. PMID- 11427855 TI - Gastric perforation after endoscopic ligation for gastric varices. PMID- 11427856 TI - Endoscopic treatment of gastritis cystica polyposa found in an unoperated stomach. PMID- 11427857 TI - Bullous colon lesions in a patient with bullous pemphigoid. PMID- 11427858 TI - Depressed type primary signet ring cell carcinoma of the colon: a case report. PMID- 11427859 TI - Aphthoid pancolitis in a patient with a verotoxin-producing Escherichia coli infection. PMID- 11427860 TI - EUS in the diagnosis of collagenous colitis. PMID- 11427861 TI - Bleeding duodenal lipoma successfully treated with endoscopic polypectomy. PMID- 11427862 TI - Directed balloon-assisted guidewire access into intrahepatic ducts. PMID- 11427863 TI - An unusual case of melanosis coli. PMID- 11427864 TI - The menu of endoscopic sedation: all-you-can-eat, combination set, a la carte, alternative cuisine, or go hungry. PMID- 11427865 TI - Extreme endoscopy: presidential address. PMID- 11427866 TI - Treatment of Zenker's diverticulum. PMID- 11427867 TI - Follow-up of Barrett's epithelium after ablation by endoscopic argon plasma coagulation. PMID- 11427869 TI - EUS and the prediction of gallbladder neoplastic polyps: are polyps of 5 to 15 mm diameter really a homogenous group? PMID- 11427871 TI - Self-expanding metal stents for the treatment of benign esophageal strictures. PMID- 11427873 TI - Is ERCP a procedure for all, the majority, or just a few endoscopists? A dilemma. PMID- 11427875 TI - The analysis of clinical studies: comparison of means, part I. PMID- 11427876 TI - Cushing's syndrome. PMID- 11427877 TI - Rethinking aspects of the training system. PMID- 11427878 TI - Finding the function buried in SAND. PMID- 11427879 TI - The double life of PX domains. PMID- 11427880 TI - Mapping out the basement membrane. PMID- 11427881 TI - ReCoGnizing methylated DNA. PMID- 11427882 TI - The structure of photosystem I. PMID- 11427883 TI - The descent of polymerization. PMID- 11427884 TI - Twenty years ago. PMID- 11427885 TI - Picture story. A DNA acrobat. PMID- 11427886 TI - Structure of the Rho-activating domain of Escherichia coli cytotoxic necrotizing factor 1. AB - Certain uropathogenic and neonatal meningitis-causing strains of Escherichia coli express a 114 kDa protein toxin called cytotoxic necrotizing factor 1 (CNF1). The toxin causes alteration of the host cell actin cytoskeleton and promotes bacterial invasion of blood-brain barrier endothelial cells. CNF1 belongs to a unique group of large cytotoxins that cause constitutive activation of Rho guanosine triphosphatases (GTPases), which are key regulators of the actin cytoskeleton. This group also includes E. coli cytotoxic necrotizing factor 2 (CNF2, 114 kDa) and dermonecrotic toxins (DNT, 159 kDa) of Bordetella spp. with related sequences occurring in Yersinia spp. Here we show that the catalytic region of CNF1 exhibits a novel protein fold as determined by its 1.83 A resolution crystal structure. The structure reveals that CNF1 has a Cys-His-main chain oxygen catalytic triad reminiscent of enzymes belonging to the catalytic triad superfamily. The position of the catalytic Cys residue at the base of a deep pocket restricts access to potential substrates and helps explain the high specificity of this and related toxins. PMID- 11427887 TI - Selected peptide extension contacts hydrophobic patch on neighboring zinc finger and mediates dimerization on DNA. AB - Protein-protein interactions often play a crucial role in stabilizing protein-DNA complexes and thus facilitate site-specific DNA recognition. We have worked to incorporate such protein-protein contacts into our design and selection strategies for short peptide extensions that promote cooperative binding of zinc finger proteins to DNA. We have determined the crystal structure of one of these fusion protein-DNA complexes. The selected peptide extension was found to mediate dimerization by reaching across the dyad axis and contacting a hydrophobic patch on the surface of the zinc finger bound to the adjacent DNA site. The peptide zinc finger protein interactions observed in this structure are similar to those of some homeodomain heterodimers. We also find that the region of the zinc finger surface contacted by the selected peptide extension corresponds to surfaces that also make key interactions in the zinc finger proteins GLI and SWI5. PMID- 11427888 TI - Structure of GSK3beta reveals a primed phosphorylation mechanism. AB - GSK3beta was identified as the kinase that phosphorylates glycogen synthase but is now known to be involved in multiple signaling pathways. GSK3beta prefers prior phosphorylation of its substrates. We present the structure of unphosphorylated GSK3beta at 2.7 A. The orientation of the two domains and positioning of the activation loop of GSK3beta are similar to those observed in activated kinases. A phosphate ion held by Arg 96, Arg 180 and Lys 205 occupies the same position as the phosphate group of the phosphothreonine in activated p38gamma, CDK2 or ERK2. A loop from a neighboring molecule in the crystal occupies a portion of the substrate binding groove. The structure explains the unique primed phosphorylation mechanism of GSK3beta and how GSK3beta relies on a phosphoserine in the substrate for the alignment of the beta- and alpha-helical domains. PMID- 11427889 TI - Interhelical hydrogen bonds in the CFTR membrane domain. AB - Critical mutations in the membrane-spanning domains of proteins cause many human diseases. We report the expression in Escherichia coli of helix-loop-helix segments of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel domain in milligram quantities. Analysis of gel migration patterns of these constructs, in conjunction with circular dichroism spectroscopy, demonstrate that a neutral-to-charged, CF-phenotypic point mutation of a hydrophobic residue (V232D) in the CFTR transmembrane (TM) helix 4 induces a hydrogen bond with neighboring wild type Gln 207 in TM helix 3. As an electrostatic crosslink within a hydrocarbon phase, such a hydrogen bond could alter the normal assembly and alignment of CFTR TM helices and/or impede their movement in response to substrate transport. Our results imply that membrane proteins may be vulnerable to loss of function through formation of membrane buried interhelical hydrogen bonds by partnering of proximal polar side chains. PMID- 11427890 TI - Computer-based redesign of a protein folding pathway. AB - A fundamental test of our current understanding of protein folding is to rationally redesign protein folding pathways. We use a computer-based design strategy to switch the folding pathway of protein G, which normally involves formation of the second, but not the first, beta-turn at the rate limiting step in folding. Backbone conformations and amino acid sequences that maximize the interaction density in the first beta-hairpin were identified, and two variants containing 11 amino acid replacements were found to be approximately 4 kcal mol-1 more stable than wild type protein G. Kinetic studies show that the redesigned proteins fold approximately 100 x faster than wild type protein and that the first beta-turn is formed and the second disrupted at the rate limiting step in folding. PMID- 11427891 TI - Unfolding individual nucleosomes by stretching single chromatin fibers with optical tweezers. AB - Single chromatin fibers were assembled directly in the flow cell of an optical tweezers setup. A single lambda phage DNA molecule, suspended between two polystyrene beads, was exposed to a Xenopus laevis egg extract, leading to chromatin assembly with concomitant apparent shortening of the DNA molecule. Assembly was force-dependent and could not take place at forces exceeding 10 pN. The assembled single chromatin fiber was subjected to stretching by controlled movement of one of the beads with the force generated in the molecule continuously monitored with the second bead trapped in the optical trap. The force displayed discrete, sudden drops upon fiber stretching, reflecting discrete opening events in fiber structure. These opening events were quantized at increments in fiber length of approximately 65 nm and are attributed to unwrapping of the DNA from around individual histone octamers. Repeated stretching and relaxing of the fiber in the absence of egg extract showed that the loss of histone octamers was irreversible. The forces measured for individual nucleosome disruptions are in the range of 20-40 pN, comparable to forces reported for RNA- and DNA-polymerases. PMID- 11427892 TI - Structural determinants of CCR5 recognition and HIV-1 blockade in RANTES. AB - Certain chemokines act as natural antagonists of human immunodeficiency virus (HIV) by blocking key viral coreceptors, such as CCR5 and CXCR4, on the surface of susceptible cells. Elucidating the structural determinants of the receptor binding and HIV-inhibitory functions of these chemokines is essential for the rational design of derivative molecules of therapeutic value. Here, we identify the structural determinants of CCR5 recognition and antiviral activity of the CC chemokine RANTES, showing that critical residues form a solvent-exposed hydrophobic patch on the surface of the molecule. Moreover, we demonstrate that the biological function is critically dependent on dimerization, resulting in the exposure of a large ( approximately 180 A2), continuous hydrophobic surface. Relevant to the development of novel therapeutic approaches, we designed a retroinverted RANTES peptide mimetic that maintained both HIV- and chemotaxis antagonistic functions. PMID- 11427893 TI - Structural basis for substrate specificities of cellular deoxyribonucleoside kinases. AB - Deoxyribonucleoside kinases phosphorylate deoxyribonucleosides and activate a number of medically important nucleoside analogs. Here we report the structure of the Drosophila deoxyribonucleoside kinase with deoxycytidine bound at the nucleoside binding site and that of the human deoxyguanosine kinase with ATP at the nucleoside substrate binding site. Compared to the human kinase, the Drosophila kinase has a wider substrate cleft, which may be responsible for the broad substrate specificity of this enzyme. The human deoxyguanosine kinase is highly specific for purine substrates; this is apparently due to the presence of Arg 118, which provides favorable hydrogen bonding interactions with the substrate. The two new structures provide an explanation for the substrate specificity of cellular deoxyribonucleoside kinases. PMID- 11427894 TI - Structure of the histone deacetylase SIRT2. AB - Sir2 is an NAD-dependent histone deacetylase that mediates transcriptional silencing at mating-type loci, telomeres and ribosomal gene clusters, and has a critical role in the determination of life span in yeast and Caenorhabditis elegans. The 1.7 A crystal structure of the 323 amino acid catalytic core of human SIRT2, a homolog of yeast Sir2, reveals an NAD-binding domain, which is a variant of the Rossmann fold, and a smaller domain composed of a helical module and a zinc-binding module. A conserved large groove at the interface of the two domains is the likely site of catalysis based on mutagenesis. Intersecting this large groove, there is a pocket formed by the helical module. The pocket is lined with hydrophobic residues conserved within each of the five Sir2 classes, suggesting that it is a class-specific protein-binding site. PMID- 11427895 TI - The SAND domain structure defines a novel DNA-binding fold in transcriptional regulation. AB - The SAND domain is a conserved sequence motif found in a number of nuclear proteins, including the Sp100 family and NUDR. These are thought to play important roles in chromatin-dependent transcriptional regulation and are linked to many diseases. We have determined the three-dimensional (3D) structure of the SAND domain from Sp100b. The structure represents a novel alpha/beta fold, in which a conserved KDWK sequence motif is found within an alpha-helical, positively charged surface patch. For NUDR, the SAND domain is shown to be sufficient to mediate DNA binding. Using mutational analyses and chemical shift perturbation experiments, the DNA binding surface is mapped to the alpha-helical region encompassing the KDWK motif. The DNA binding activity of wild type and mutant proteins in vitro correlates with transcriptional regulation activity of full length NUDR in vivo. The evolutionarily conserved SAND domain defines a new DNA binding fold that is involved in chromatin-associated transcriptional regulation. PMID- 11427896 TI - Crystal structure and mutational analysis of a perlecan-binding fragment of nidogen-1. AB - Nidogen, an invariant component of basement membranes, is a multifunctional protein that interacts with most other major basement membrane proteins. Here, we report the crystal structure of the mouse nidogen-1 G2 fragment, which contains binding sites for collagen IV and perlecan. The structure is composed of an EGF like domain and an 11-stranded beta-barrel with a central helix. The beta-barrel domain has unexpected similarity to green fluorescent protein. A large surface patch on the beta-barrel is strikingly conserved in all metazoan nidogens. Site directed mutagenesis demonstrates that the conserved residues are involved in perlecan binding. PMID- 11427897 TI - Structure of 4-diphosphocytidyl-2-C- methylerythritol synthetase involved in mevalonate- independent isoprenoid biosynthesis. AB - The YgbP protein of Escherichia coli encodes the enzyme 4-diphosphocytidyl-2-C methylerythritol (CDP-ME) synthetase, a member of the cytidyltransferase family of enzymes. CDP-ME is an intermediate in the mevalonate-independent pathway for isoprenoid biosynthesis in a number of prokaryotic organisms, algae, the plant plastids and the malaria parasite. Because vertebrates synthesize isoprenoid precursors using a mevalonate pathway, CDP-ME synthetase and other enzymes of the mevalonate-independent pathway for isoprenoid production represent attractive targets for the structure-based design of selective antibacterial, herbicidal and antimalarial drugs. The high-resolution structures of E. coli CDP-ME synthetase in the apo form and complexed with both CTP-Mg2+ and CDP-ME-Mg2+ reveal the stereochemical principles underlying both substrate and product recognition as well as catalysis in CDP-ME synthetase. Moreover, these complexes represent the first experimental structures for any cytidyltransferase with both substrates and products bound. PMID- 11427898 TI - Neural stem cell: biology and prospects of neurotransplantation. PMID- 11427899 TI - Blockade of hyperpolarizing currents produces a dose-dependent effect on heart rate. AB - Intravenous injection of ZD 7288, a new specific hyperpolarizing current blocker, dose-dependently reduces heart rate in adult rats. The autonomic nervous system modulates changes in heart rate caused by hyperpolarizing currents. PMID- 11427900 TI - Effect of ACTH on pain sensitivity in rats. AB - Systemic administration of ACTH to rats with normal hormone production induced a rapid (started 3 min postinjection) and long-term (persisted 30 min) elevation of pain threshold. Complete inhibition of glucocorticoid production shortened the duration of ACTH-induced analgesia to 15 min. The biphasic effect of ACTH on pain sensitivity is probably mediated by short-term glucocorticoid-independent and long-term glucocorticoid-dependent mechanisms. PMID- 11427901 TI - Effect of transplantation of embryonic nervous tissue on reorganization of interneuronal relationships after mechanical damage to sensorimotor cortex. AB - The effect of implanted embryonic nervous tissue on restoration of axonal connections in the cerebral cortex after mechanical injury was studied on albino rats using fluorescent lipophilic probe DiI (1,1'-dioctadecyl-3,3,3',3' tetramethyl-indocarbocyanine perchlorate) and confocal laser scanning microscope. Implantation of embryonic tissue to damaged area promotes the growth of axons through the transplant to adjacent tissue. The damaged area is impenetrable for axons growing without implantation. PMID- 11427902 TI - Acute bradyarrhythmia induced by occlusion of the posterior interventricular branch of the right coronary artery. AB - Ligation of the posterior interventricular branch of the right coronary artery in rats induced bradyarrhythmia similar by its pathophysiological mechanisms to bradyarrhythmias developed in humans during acute ischemia of the posterior cardiac wall. The type and severity of arrhythmia and conduction disturbances, their latency and duration, and correlation with the volume of damaged myocardial tissue were determined. The efficacy and safety of the use of methylxanthines during acute myocardial ischemia was proved. PMID- 11427903 TI - Urine antipyrine metabolites in rats with different resistance to hypoxia subjected to cold stress. AB - We studied antipyrine metabolism in rats with different resistance to hypoxia during adaptation to cold stress. Changes in the concentrations of some antipyrine metabolites at low temperature were associated with individual resistance to hypoxia. In low-resistant rats, antipyrine metabolism was suppressed from day 5 of cold exposure to day 3 of the recovery period. In highly resistant rats, antipyrine metabolism was inhibited on day 3 of cold exposure, but returned to normal on day 3 of the recovery period. PMID- 11427904 TI - Effect of xymedon on cell survival in the system sensory neuron Schwann cell. AB - Pyrimidine derivative xymedon inhibits neuronal death in L4-L5 spinal ganglia 30 days after ligation of rat sciatic nerve. After treatment with xymedon the number of neurons on the operated side decreased by 22.1% compared to that on the contralateral side, while in the control group this parameter decreased by 28.7%. At the same terms, the number of Schwann cells on the operated side after xymedon injection increased by 27.7% in comparison with that on the contralateral side, while in the control group this parameter decreased by 57.3%. PMID- 11427905 TI - Maintenance of homeostasis of endogenous ethanol as a method for the therapy of alcoholism. AB - We propose a new method for the therapy of alcoholism based on maintenance of homeostasis of endogenous ethanol and inhibition of alcohol dehydrogenase with emetine. After the standard antialcohol therapy, activity of this enzyme remained high or even increased, and pathological alcohol addiction also increased. Emetine normalized activity of alcohol dehydrogenase and suppressed pathological alcohol addiction. After this therapy more than 50% patients achieved stable remissions from alcoholism over 1 year, which indicated high efficiency of the proposed method. PMID- 11427906 TI - Oxidative modification of plasma proteins during hypothermia and after dalargin administration. AB - Short-term and prolonged (3 h) moderate (30 degrees C) hypothermia intensified oxidative modification of plasma proteins, while deep hypothermia (20 degrees C) decreased the intensity of this process to a control level. Preliminary intraperitoneal injection of dalargin had practically no effect on oxidative modification of plasma proteins during moderate hypothermia. PMID- 11427907 TI - Effect of disaggregation on calpain activity in explants from rat thyroid glands. AB - We studied the regulation of calpains in explants from rat thyroid glands. Collagenase disaggregation decreased proteinase activity in thyrocytes. It was hypothesized that this effect is mediated by heterotrimeric G proteins, protein kinase C, and tyrosine kinase, but not via Ca(2+) and cAMP-dependent signal pathway. PMID- 11427908 TI - Mechanism of therapeutic effect of low-intensity infrared laser radiation. AB - The effect of infrared low-intensity laser irradiation on functional activity of blood polymorphonuclear leukocytes was studied in vitro. A dose-dependent priming of polymorphonuclear leukocytes induced by infrared low-intensity laser irradiation was demonstrated. Similar effects were also observed in the presence of the photosensitizer photosense. PMID- 11427909 TI - Effect of ionic carbohydrate-containing biopolymers on complement activation. AB - We studied in vitro effects of charged polysaccharides on the classic and alternative pathways of complement activation. The complement system was affected by substances having different charges. Our findings suggest that the conformation of polysaccharide molecules, but not their charge, plays a primary role in the interaction with C1 and C3 complement components followed by initiation of cascade enzymatic reactions. PMID- 11427910 TI - Bioactive 5-reduced 16 alpha,17 alpha-cyclohexanoprogesterone derivatives weakly interact with proteins of the soluble uterine fraction. AB - We studied competitive activities of 16 alpha,17 alpha-cyclohexano-5 alpha- and 5 beta-dihydroprogesterone in replacing(3)H-progesterone and(3)H-16 alpha,17 alpha cyclohexano-6 alpha-methylprogesterone from protein complexes. Direct binding of(3)H-5-reduced derivatives with proteins of soluble fractions from rat and rabbit uteri was also assayed. C(d) values for 5-reduced derivatives were in the micro- or submicromolar range. The data suggest that biological effects of these analogues are not mediated via soluble uterine receptors. PMID- 11427911 TI - Effect of morphine on the number and branching of astrocytes in various regions of rat brain. AB - We studied the effects of morphine in high doses on astrocytes in the nucleus accumbens, lateral septum, and caudate nucleus of rat brain. Activation of astrocytes in the nucleus accumbens and lateral septal nucleus was manifested in hyperplasia and elongation of astrocyte processes. In the caudate nucleus, the total length of astrocyte processes and branching of individual astrocytes decreased. PMID- 11427912 TI - Endogenous cannabinoid anandamide increases heart resistance to arrhythmogenic effects of epinephrine: role of CB(1) and CB(2) receptors. AB - Intravenous injection of 10 mg/kg anandamide reduces the incidence and duration of epinephrine-induced arrhythmias in rats. SR141716A and SR144528, antagonists of cannabinoid receptor I and II did not abolish the antiarrhythmic effect of anandamide. These data suggest that the antiarrhythmic effect of anandamide is nonspecific or mediated via unknown cannabinoid receptors, but not associated with activation of cannabinoid receptors I and II. PMID- 11427913 TI - Analgesic effect of olipiphate on mouse model of chemical stimulation of peritoneum. AB - We studied the analgesic effect of olipiphate, a product of lignin, against writhing provoked by intraperitoneal injection of acetic acid. Paracetamol was used as the reference drug. Both agents dose-dependently decreased the number of motor reactions caused by the irritant. Olipiphate possessed analgesic activity and efficiency comparable with those of paracetamol, but produced a more long lasting effect. PMID- 11427914 TI - Regulatory effects of Ribotim on functional activity of neutrophils and wound healing during experimental burn trauma. AB - We studied the effects of ribonucleic immunotropic preparation Ribotim obtained from the thymus on functional activity of peripheral blood neutrophils and wound healing after burn trauma. Immunotherapy normalized immunological parameters, attenuated demarcation inflammation in burn wound, and promoted epithelialization of the wound surface preserving skin structures. PMID- 11427915 TI - Abnormalities of human genetic apparatus manifested in leukocyte proteins in psoriasis. AB - The content of 53K/H2A and 43K/H2A proteins in leukocytes of patients with psoriasis, newborns, and healthy donors was studied. In all patients one or both parameters differed from normal. Mean values and distribution of 53K/H2A in healthy siblings of psoriatic proband significantly differed from those in newborns and donors. It was concluded that 53K/H2A is a marker of pathological changes in the genome responsible for predisposition to psoriasis. This marker can be revealed in peripheral blood leukocytes before skin manifestations of the disease. PMID- 11427916 TI - Role of alpha(2A)-adrenoceptors of locus coeruleus in regulation of plasma corticosterone content in male rats. AB - Administration of specific oligonucleotide selectively inhibiting alpha(2A) adrenoceptor gene expression into the locus coeruleus of male rats for 3 days activated the hypothalamic-pituitary-adrenal system, which was manifested in a rise of blood plasma corticosterone content in rats with normal and hypertrophied (after castration) adrenal glands. These data indicate that alpha(2A) adrenoceptors of the locus coeruleus are involved in the regulation of basal plasma corticosterone content. PMID- 11427917 TI - Modification of hemostatic status improves antitumor efficiency of photodynamic therapy. AB - Complex treatment with acetylsalicylic acid, heparin, and dexamethasone improves the efficiency of photodynamic therapy in rats with myosarcoma-I. PMID- 11427918 TI - Induction of blebbing in Ehrlich ascitic adenocarcinoma cells during in vitro hyperthermia. AB - We studied the effect of in vitro hyperthermia on induction of blebbing in cultured Ehrlich ascitic adenocarcinoma cells. Hyperthermia (42-48 degrees C) promotes induction of blebbing in cell culture, but this induction is reversible, and cessation of hyperthermia leads to almost complete recovery of the morphological composition of cell suspension. PMID- 11427919 TI - Metabolic interrelations between rat brain lipids during geomagnetic perturbations. AB - Lipid components in various brain structures undergo different changes during geomagnetic perturbations. These changes involve synthetase, hydrolase, and transferase biotransformations of lipids. These changes reflect differences in metabolic activities in the left and right cerebral hemispheres and the development of biochemical cerebral asymmetry under the effect of natural stress factors. Changes in lipid metabolism probably reduce adaptive capacities of the organism. PMID- 11427920 TI - Effect of audiovisual stimulation on heart rhythm variability. AB - Presentation of visual and acoustic stimuli at a rate of 2-30 Hz modulated variability of the heart rhythm in volunteers. Potentation of vagal influences and a decrease in the contribution of ergotropic systems to the regulation of the heart rate were noted after 10 sessions of audiovisual stimulation. These changes were accompanied by a pronounced decrease of individual and reactive anxiety. PMID- 11427921 TI - Suppressed production of transforming growth factor-beta(2)mRNA in endometrium of women with polycystic ovary syndrome. AB - Production of transforming growth factor-beta(2)mRNA in the endometrium of women with polycystic ovary syndrome decreased compared to normal and this decrease directly depends on the duration of anovulatory period (from 3 weeks to 4 months). Low production of transforming growth factor-beta(2)mRNA probably contributes to the development of endometrial hyperplasia in women with polycystic ovary syndrome. PMID- 11427922 TI - Effects of various modes of sonication with low frequency ultrasound on in vitro survival of human tumor cells. AB - The effect of low frequency ultrasound in the cavitation (5-15 sec exposure) and subcavitation (5-210 sec exposure) modes on in vitro survival of cultured human tumor cells was studied. Analysis of the dose-effect curves and mathematical estimation of the effects low-frequency ultrasonication in the subcavitation mode on cell membranes helped to choose time intervals of tumor cell sonication ensuring potentiation of the cytostatic effects. PMID- 11427923 TI - Detection of mutant k-ras sequences in the urine of cancer patients. AB - DNA fragments from apoptotic cells crossing the renal barrier retain their matrix functions, which allows PCR identification of mutant sequences in excreted DNA. We investigated the possibility of detecting k-ras mutations in urinary DNA of tumor patients (colon cancer). In some patients with k-ras codon 12 mutations in tumor cell DNA the same changes were detected in the urinary DNA. The possibility of using this approach for early diagnosis and monitoring of tumors is discussed. PMID- 11427924 TI - Immunohistochemical and morphometric analysis of effects of vilon and epithalon on functional morphology of radiosensitive organs. AB - Studies of the effects of vilon and epithalon on functional morphology of the thymus, spleen, and duodenum in intact rats and rats exposed to single whole-body gamma-irradiation in a dose of 6 Gy showed that vilon stimulated proliferative activity of thymocytes and enhanced proliferative potential of stem cells in the intestine, thus stimulating the postradiation recovery of critical organs. Epithalon decelerated metabolic processes in the duodenal mucosa and suppressed hemopoiesis and lymphopoiesis in the spleen. PMID- 11427925 TI - Ultrastructure of the contractile apparatus in cardiomyocytes during regenerative and plastic insufficiency of the myocardium. AB - Lytic changes in cardiomyocyte myofibrils constituting the morphological basis of contractile insufficiency were found in Wistar rats with regenerative and plastic myocardial insufficiency 3 h after daunomycin administration. Myofibrils became less dense, empty spaces appeared in many sarcomeres, sometimes total lysis of myofilaments within the sarcomere was noted. These changes were most pronounced in the perinuclear zone. Intracellular regeneration of cardiomyocytes was characterized by disorientation of newly formed myofibrils in relation to the long axis of muscle fibers and preserved myofibrils. Progressive inhibition of protein synthesis, lysis of myofibrils, and focal degradation of the sarcoplasm caused apoptotic death of some cardiomyocytes. PMID- 11427926 TI - Ultrastructural organization of Helicobacter pylori under natural conditions and during ex vivo culturing. AB - Ultrastructure of Helicobacter pylori were studied by transmission electron microscopy in freshly isolated and ex vivo cultured biopsy specimens of gastric antral mucosa from patients with duodenal ulcer. PMID- 11427927 TI - Albumin fluorescent test in surgical stress. AB - Albumin fluorescent test showed that serum albumin concentration was normal in patients with uncomplicated inguinal hernia before surgery, but albumin binding centers were considerably changed. Surgery also modified binding centers, but these changes were less pronounced than before the operation. It was assumed that long-term psychic distress before surgery is responsible for preoperative changes in albumin molecule. PMID- 11427937 TI - Optical method for the determination of the oxygen-transfer capacity of small bioreactors based on sulfite oxidation. AB - The growth of microorganisms may be limited by operating conditions which provide an inadequate supply of oxygen. To determine the oxygen-transfer capacities of small-scale bioreactors such as shaking flasks, test tubes, and microtiter plates, a noninvasive easy-to-use optical method based on sulfite oxidation has been developed. The model system of sodium sulfite was first optimized in shaking flask experiments for this special application. The reaction conditions (pH, buffer, and catalyst concentration) were adjusted to obtain a constant oxygen transfer rate for the whole period of the sulfite oxidation reaction. The sharp decrease of the pH at the end of the oxidation, which is typical for this reaction, is visualized by adding a pH dye and used to measure the length of the reaction period. The oxygen-transfer capacity can then be calculated by the oxygen consumed during the complete stoichiometric transformation of sodium sulfite and the visually determined reaction time. The suitability of this optical measuring method for the determination of oxygen-transfer capacities in small-scale bioreactors was confirmed with an independent physical method applying an oxygen electrode. The correlation factor for the maximum oxygen transfer capacity between the chemical model system and a culture of Pseudomonas putida CA-3 was determined in shaking flasks. The newly developed optical measuring method was finally used for the determination of oxygen-transfer capacities of different types of transparent small-scale bioreactors. PMID- 11427938 TI - Probing the performance limits of the Escherichia coli metabolic network subject to gene additions or deletions. AB - An optimization-based procedure for studying the response of metabolic networks after gene knockouts or additions is introduced and applied to a linear flux balance analysis (FBA) Escherichia coli model. Both the gene addition problem of optimally selecting which foreign genes to recombine into E. coli, as well as the gene deletion problem of removing a given number of existing ones, are formulated as mixed-integer optimization problems using binary 0-1 variables. The developed modeling and optimization framework is tested by investigating the effect of gene deletions on biomass production and addressing the maximum theoretical production of the 20 amino acids for aerobic growth on glucose and acetate substrates. In the gene deletion study, the smallest gene set necessary to achieve maximum biomass production in E. coli is determined for aerobic growth on glucose. The subsequent gene knockout analysis indicates that biomass production decreases monotonically, rendering the metabolic network incapable of growth after only 18 gene deletions. In the gene addition study, the E. coli flux balance model is augmented with 3,400 non-E. coli reactions from the KEGG database to form a multispecies model. This model is referred to as the Universal model. This study reveals that the maximum theoretical production of six amino acids could be improved by the addition of only one or two genes to the native amino acid production pathway of E. coli, even though the model could choose from 3,400 foreign reaction candidates. Specifically, manipulation of the arginine production pathway showed the most promise with 8.75% and 9.05% predicted increases with the addition of genes for growth on glucose and acetate, respectively. The mechanism of all suggested enhancements is either by: 1) improving the energy efficiency and/or 2) increasing the carbon conversion efficiency of the production route. PMID- 11427939 TI - Assessment of near-infrared spectral information for rapid monitoring of bioprocess quality. AB - Access to real-time process information is desirable for consistent and efficient operation of bioprocesses. Near-infrared spectroscopy (NIRS) is known to have potential for providing real-time information on the quantitative levels of important bioprocess variables. However, given the fact that a typical NIR spectrum encompasses information regarding almost all the constituents of the sample matrix, there are few case studies that have investigated the spectral details for applications in bioprocess quality assessment or qualitative bioprocess monitoring. Such information would be invaluable in providing operator level assistance on the progress of a bioprocess in industrial-scale productions. We investigated this aspect and report the results of our investigation. Near infrared spectral information derived from scanning unprocessed culture fluid (broth) samples from a complex antibiotic production process was assessed for a data set that incorporated bioprocess variations. Principal component analysis was applied to the spectral data and the loadings and scores of the principal components studied. Changes in the spectral information that corresponded to variations in the bioprocess could be deciphered. Despite the complexity of the matrix, near-infrared spectra of the culture broth are shown to have valuable information that can be deconvoluted with the help of factor analysis techniques such as principal component analysis (PCA). Although complex to interpret, the loadings and score plots are shown to offer potential in process diagnosis that could be of value in the rapid assessment of process quality, and in data assessment prior to quantitative model development. PMID- 11427940 TI - Bromoperoxidase activity in microplantlet suspension cultures of the macrophytic red alga Ochtodes secundiramea. AB - Bromoperoxidase is an enzyme found in marine macroalgae that catalyzes the bromination of organic substrates. Photosynthetic microplantlet suspension cultures derived from the macrophytic red alga Ochtodes secundiramea were shown to possess bromoperoxidase. The optimum pH for O. secundiramea bromoperoxidase activity in cell-free extracts was 6.0, and the half-saturation constant for bromination of the exogeneous substrate monochlorodimedone (MCD) was 18 microM. O. secundiramea microplantlets were cultivated in a bubble-column photobioreactor at an incident light intensity of 38 microE x m(-2) x s(-1) (71% of light saturated photosynthesis, 10:14 light:dark photoperiod), and the kinetics of cell growth and bromoperoxidase production were followed. At these conditions, the specific growth rate was 0.052 x day(-1). The lowest specific bromoperoxidase activity of 0.3 micromol MCD x g(-1) cell x min(-1) occurred during the midexponential phase of growth, and then increased steeply to 1.9 micromol MCD x g(-1) cell x min(-1) during the late stationary phase, suggesting that bromoperoxidase production was part of secondary metabolism. The estimated bromoperoxidase content in the cell mass at late stationary phase was 67 microg x g(-1) dry cell mass, demonstrating that bioreactor production of marine bromoperoxidase is feasible. PMID- 11427941 TI - Effect of media composition on yield values of bacteria growing on binary and ternary substrate mixtures in continuous culture. AB - Pseudomonas aeruginosa 142 and a presumed variant were grown axenically in chemostats on salicylate/benzoate or salicylate/glucose binary feeds. Each substrate was supplied at 2, 10, 50, 90, 98, or 100% of the total energy flux. Two experiments were also run with ternary mixtures using the same substrates. Aliquots were transferred to fed-batch reactors receiving the same substrates at the same specific rates as the chemostat, but with one substrate radiolabeled with 14C. Radiolabel incorporated into biomass, 14CO2, and soluble microbial products over a period of 8 minutes was used to establish the biomass yield, CO2 yield, and product yield, respectively, associated with a given substrate. The effect of the percent substrate in the feed on the yields depended on the pair of substrates supplied. When benzoate comprised 50% or more of the applied substrate in salicylate/benzoate feeds, the fraction of benzoate in the feed had a small effect on the yield values associated with benzoate. However, when benzoate constituted 2% or 10% of the feed, CO2 yields were lower, biomass yields were slightly lower, and product yields were higher. In contrast, the percent of salicylate in the feed had little effect on any of the salicylate yields for cells growing on the salicylate/benzoate feeds. When salicylate was mixed with glucose, the yields associated with salicylate behaved quite differently. Biomass and CO2 yields were lower and product yields higher when salicylate was 2% or 10% of the feed than when it was higher. In the same substrate mixtures, glucose based biomass yields were higher and CO2 yields were lower when glucose constituted 2% or 10% of the feed but were constant for higher percentages. The results suggest that the fate of a substrate is relatively independent of the feed composition as long as the substrate in question constitutes a significant percentage of the mixture. Thus, in those situations the assumption of a constant biomass yield in multicomponent substrate modeling is justified. However, when a given substrate constitutes a small percentage of the feed, significant changes in yield may occur. PMID- 11427942 TI - Novel kinetic analysis of enzymatic dipeptide synthesis: effect of pH and substrates on thermolysin catalysis. AB - The point of maximum activity is specific to a particular substrate-enzyme system but may vary with different substrates and the same enzyme. The specificity of enzymes has, however, been generally reported only at their "optimal" pH. In this article, we introduce the Michaelis-Menten equation taking pH into account, and apply it to the pH-activity profile of the thermolysin-catalyzed dipeptide synthesis. It has been reported to date that the pH-activity profile of thermolysin follows a bell-shaped curve with a maximal activity at or near pH 7.0. The profiles obtained in this study, however, indicated that the optimal pH varied from 5.8 (for F-AspPheOMe) to 7.3 (for Z-ArgPheOMe), and the order of thermolysin activity was greatly dependent on the pH of reaction media. We have succeeded in evaluating the substrates-induced change of the dissociation states of the active site of thermolysin using the hydrophobicity of substrates. We have obtained apparent kinetic parameters which are independent of the pH of reaction media. The apparent specificity of thermolysin which were independent of pH of the reaction media was in order L-Leu > L-Asp > L-Arg > L-Ala > L-Gly > L-Val and Z > Boc = F at P1 and P2 positions, respectively. PMID- 11427943 TI - Replacing factor-dependency with that for lysozyme: affordable culture of IL-6 dependent hybridoma by transfecting artificial cell surface receptor. AB - Cytokines and growth factors are indispensable for the propagation and maintenance of factor-dependent mammalian cells. However, cytokines are often so expensive that the use of factor-dependent cells for industrial applications such as protein production is often not practical. Based on our previous design of a binary hen egg lysozyme (HEL)-specific receptor composed of portions of the anti HEL antibody and the erythropoietin receptor, a new pair of chimeric receptors having the intracellular domain of gp130 were made and transfected to an interleukin-6 (IL-6)-dependent hybridoma, 7TD1. The clone expressing the two new receptors showed clear HEL dose-dependent cell growth and monoclonal antibody production in both serum-based and serum-free media without IL-6. These results establish the feasibility of applying receptor design to tailor cells for the inexpensive induction of cell growth for the purpose of producing therapeutic products. PMID- 11427944 TI - Mechanistic study of microbial control of hydrogen sulfide production in oil reservoirs. AB - Microbial control of biogenic production of hydrogen sulfide in oil fields was studied in a model system consisting of pure cultures of the nitrate-reducing, sulfide-oxidizing bacterium (NR-SOB) Thiomicrospira sp. strain CVO and the sulfate-reducing bacterium (SRB) Desulfovibrio sp. strain Lac6, as well as in microbial cultures enriched from produced water of a Canadian oil reservoir. The presence of nitrate at concentrations up to 20 mM had little effect on the rate of sulfate reduction by a pure culture of Lac6. Addition of CVO imposed a strong inhibition effect on production of sulfide. In the absence of added nitrate SRB we were able to overcome this effect after an extended lag phase. Simultaneous addition of CVO and nitrate stopped the production of H2S immediately. The concentration of sulfide decreased to a negligible level due to nitrate-dependent sulfide oxidation activity of CVO. This was not prevented by raising the concentration of Na-lactate, the electron donor for sulfate reduction. Similar results were obtained with enrichment cultures. Enrichments of produced water with sulfide and nitrate were dominated by CVO, whereas enrichments with sulfate and Na-lactate were dominated by SRB. Addition of an NR-SOB enrichment to an SRB enrichment inhibited the production of sulfide. Subsequent addition of sufficient nitrate caused the sulfide concentration to drop to zero. A similar response was seen in the presence of nitrate alone, although after a pronounced lag time, it was needed for emergence of a sizable CVO population. The results of the present study show that two mechanisms are involved in microbial control of biogenic sulfide production. First, addition of NR-SOB imposes an inhibition effect, possibly by increasing the environmental redox potential to levels which are inhibitory for SRB. Second, in the presence of sufficient nitrate, NR-SOB oxidize sulfide, leading to its complete removal from the environment. Successful microbial control of H2S in an oil reservoir is crucially dependent on the simultaneous presence of NR-SOB (either indigenous population or injected) and nitrate in the environment. PMID- 11427945 TI - Flow cytometry: an improved method for the selection of highly productive gene amplified CHO cells using flow cytometry. AB - In previous work, we clarified the relationship between the productivity and stability of gene-amplified cells and the location of the amplified gene. The location of the amplified gene enabled us to classify resistant cells into two types. One type of resistant cell group, in which the amplified genes were observed near the telomeric region, was named the "telomere type." The other type of cell group, in which the amplified genes were observed in other chromosomal regions, was named the "other type." The phenotypes of these two types of cells are very different. In this experiment, using a fluorescein isothiocyanate labeled methotrexate (F-MTX) reagent with flow cytometry, we were easily able to distinguish between highly productive cells and the other types of cells. The level of fluorescence differed according to the difference in resistance to MTX. Based on this new finding, highly productive gene-amplified cells could be isolated from heterogeneous gene-amplified cell pools more easily than by the method of limiting-dilution assay. The limiting-dilution method requires several months to obtain highly productive gene-amplified cells, while our flow-cytometry based method of selection requires only a few weeks. PMID- 11427946 TI - Assessment of effects of experimental imprecision on optimized biochemical systems. AB - Metabolic pathways may be optimized with S-system models that prescribe profiles of control variables leading to optimal output while keeping metabolites and enzyme activities within predefined ranges. Monte Carlo simulations show how much the yield and the corresponding metabolite concentrations would be affected by inaccuracies in the experimental implementation of the prescribed profiles. For a recent model of citric acid production in Aspergillus niger, the yield is roughly normally distributed, whereas the distributions of metabolite concentrations differ greatly in shape and statistical characteristics. Even moderate inaccuracies may lead to constraint violations, which appear to be correlated with high logarithmic gains. PMID- 11427947 TI - Use of odds ratio or relative risk to measure a treatment effect in clinical trials with multiple correlated binary outcomes: data from the NINDS t-PA stroke trial. AB - In clinical trials, when a single outcome is not sufficient to describe the underlying concept of interest, it may be necessary to compare treatment groups on multiple correlated outcomes. A global test based on a logit link function provides an estimate of the odds ratio for assessing a common treatment effect among correlated binary outcomes. In this paper we extend the use of generalized estimating equations (GEE) to calculate a common relative risk from correlated binary outcomes based on a log link function. In the context of global tests, we discuss the equivalence and difference between logit and log links and their estimates. We also derive a formula for calculating a common risk difference between two treatment groups based on multiple correlated binary outcomes with categorical covariates, assuming the asymptotic equivalency between the logit and log-linear links. We discuss the statistical tools to be used in choosing between the logit and log links when models on different links yield contrasting results. Examples using data from the NINDS t-PA Stroke Trials are provided. We conclude, in a study of correlated binary outcomes, that the choice of the logit or log link could be based on a comparison of goodness-of-link. PMID- 11427948 TI - Group sequential test strategies for superiority and non-inferiority hypotheses in active controlled clinical trials. AB - In a group sequential active controlled clinical trial, the study hypothesis may be a superiority hypothesis that an experimental treatment is more effective than the active control therapy or a non-inferiority hypothesis that the treatment is no worse than the active control within some non-inferiority range. When it is necessary to plan for testing the superiority and the non-inferiority hypotheses, we propose an adaptive group sequential closed test strategy by which the sample size is planned for testing superiority and is to be increased for showing non inferiority given that it is deemed more plausible than superiority based on the observed sample path during the course of the trial. The proposed adaptive test strategy is valid in terms of having the type I error probability maintained at the targeted alpha level for both superiority and non-inferiority. It has power advantage or sample size saving over the traditional group sequential test designed for testing either superiority only or non-inferiority only. PMID- 11427949 TI - An improved method of evaluating drug effect in a multiple dose clinical trial. AB - In drug development, finding an optimal dose is normally carried out in a phase II trial. A phase III trial will then be conducted to demonstrate that the selected dose is efficacious and safe. As choosing a dose from the phase II trial which has the highest observed response rate could overestimate the true response rate of the selected dose, the data from the phase II study cannot be simply pooled with the data from the phase III study in a final analysis. Therefore, a solution to the overestimation problem needs to be found so that the information obtained from phase II dose finding clinical trials can appropriately be combined with the data in the phase III study. In this paper, the potential overestimation in a multiple dose clinical trial is assessed and a method for correcting this bias is proposed. Simulations show that stepwise over-correction, the proposed method, is better than methods such as Bonferroni's procedure. PMID- 11427950 TI - Simultaneous modelling of operative mortality and long-term survival after coronary artery bypass surgery. AB - Typical analyses of lifetime data treat the time to death or failure as the response variable and use a variety of modelling strategies such as proportional hazards or fully parametric, to investigate the relationship between the response and covariates. In certain circumstances it may be more natural to view the distribution of the response variable as consisting of two or more parts since the survival curve appears segmented. This article addresses such a scenario and we propose a model for simultaneously investigating the effects of covariates over the two segments. The model is an analogue of that proposed by Lambert for zero-inflated Poisson regression. The application is central to the model development and is concerned with survival after coronary artery bypass surgery. Here operative mortality, defined as death within 30 days after surgery, and long term mortality, are viewed as distinct outcomes. For the application considered, the survivor function displays much steeper descent during the first 30 days after surgery, that is, for operative mortality, than after this period. An investigation of the effects of covariates on operative and long-term mortality after coronary artery bypass surgery illustrates the usefulness of the proposed model. PMID- 11427951 TI - Meta-analysis of binary data: which within study variance estimate to use? AB - We applied a mixed effects model to investigate between- and within-study variation in improvement rates of 180 schizophrenia outcome studies. The between study variation was explained by the fixed study characteristics and an additional random study effect. Both rate difference and logit models were used. For a binary proportion outcome p(i) with sample size n(i) in the ith study, (circumflexp(i)(1-circumflexp(i))n)(-1) is the usual estimate of the within-study variance sigma(i)(2) in the logit model, where circumflexpi) is the sample mean of the binary outcome for subjects in study i. This estimate can be highly correlated with logit(circumflexp(i)). We used (macronp(i)(1-macronp)n(i))(-1) as an alternative estimate of sigma(i)(2), where macronp is the weighted mean of circumflexp(i)'s. We estimated regression coefficients (beta) of the fixed effects and the variance (tau(2)) of the random study effect using a quasi likelihood estimating equations approach. Using the schizophrenia meta-analysis data, we demonstrated how the choice of the estimate of sigma(2)(i) affects the resulting estimates of beta and tau(2). We also conducted a simulation study to evaluate the performance of the two estimates of sigma(2)(i) in different conditions, where the conditions vary by number of studies and study size. Using the schizophrenia meta-analysis data, the estimates of beta and tau(2) were quite different when different estimates of sigma(2)(i) were used in the logit model. The simulation study showed that the estimates of beta and tau(2) were less biased, and the 95 per cent CI coverage was closer to 95 per cent when the estimate of sigma(2)(i) was (macronp(1-macronp)n(i))(-1) rather than (circumflexp(i)(1-circumflexp)n(i))(-1). Finally, we showed that a simple regression analysis is not appropriate unless tau(2) is much larger than sigma(2)(i), or a robust variance is used. PMID- 11427952 TI - Regression analysis of group testing samples. AB - This paper develops a general regression methodology that relates the group testing responses to individual covariate information. It can be used to study samples from a group testing procedure and to deal with a wide range of regression problems. A detailed illustration of the methodology is provided for a group testing procedure proposed by Gastwirth and Hammick. To demonstrate the utility of the method, simulation studies are performed on an HIV antibody testing data set published by Nusbacher et al. PMID- 11427953 TI - Sample size calculations for clustered binary data. AB - In this paper we propose a sample size calculation method for testing on a binomial proportion when binary observations are dependent within clusters. In estimating the binomial proportion in clustered binary data, two weighting systems have been popular: equal weights to clusters and equal weights to units within clusters. When the number of units varies cluster by cluster, performance of these two weighting systems depends on the extent of correlation among units within each cluster. In addition to them, we will also use an optimal weighting method that minimizes the variance of the estimator. A sample size formula is derived for each of the estimators with different weighting schemes. We apply these methods to the sample size calculation for the sensitivity of a periodontal diagnostic test. Simulation studies are conducted to evaluate a finite sample performance of the three estimators. We also assess the influence of misspecified input parameter values on the calculated sample size. The optimal estimator requires equal or smaller sample sizes and is more robust to the misspecification of an input parameter than those assigning equal weights to units or clusters. PMID- 11427954 TI - A note on the distribution of the number of vaccinated infected under non-random mixing conditions. AB - A common situation in vaccine efficacy (VE) estimation is dealing with non randomly mixing populations, which may subject vaccinated and unvaccinated individuals to a different infection pressure. These conditions may lead to a bias in VE estimates. The derivation of the statistical distribution of the number of vaccinated and infected out of a sample of n infections in a VE trial is essential to develop estimates and their properties. For randomly mixing populations, it has been shown recently that this follows a hypergeometric distribution for 'all/nothing' vaccines, whereas it is a non-central hypergeometric distribution for 'leaky' ones. Here it is shown that these distributions still hold under non-random mixing conditions, provided that mixing preferences and contact rates are independent of vaccination status. These conditions are met when vaccine and placebo are randomized. PMID- 11427955 TI - Some issues in resolution of diagnostic tests using an imperfect gold standard. AB - As a subject's true disease status is seldom known with certainty, it is necessary to compare the performance of new diagnostic tests with those of a currently accepted but imperfect 'gold standard'. Errors made by the gold standard mean that the sensitivity and specificity calculated for the new test are biased, and do not correctly estimate the new method's sensitivity and specificity. The traditional approach to this problem was 'discrepant resolution', in which the subjects for whom the two methods disagreed were subjected to a third 'resolver' test. Recent work has pointed out that this does not automatically solve the problem. A sounder approach goes beyond the discordant test results and tests at least some of the subjects with concordant results with the resolver also. This leaves some issues unresolved. One is the basic question of the direction of biases in various estimators. We point out that this question does not have a simple universal answer. Another issue, if one is to test a sample of the subjects with concordant results rather than all cases, is how to compute estimates and standard errors of the measures of test performance, notably sensitivity and specificity of the test method relative to the resolver. Expressions for these standard errors are given and illustrated with a numeric example. It is shown that using just a sample of subjects with concordant results may lead to great savings in assays. The design issue of how many concordant cells to test depends on the numbers of concordants and discordants. The formulae given show how to evaluate impact of different choices for these numbers and hence settle on a design that gives the required precision of estimates. PMID- 11427956 TI - Estimating HIV incidence rates from age prevalence data in epidemic situations. AB - We present a method of estimating HIV incidence rates in epidemic situations from data on age-specific prevalence and changes in the overall prevalence over time. The method is applied to women attending antenatal clinics in Hlabisa, a rural district of KwaZulu/Natal, South Africa, where transmission of HIV is overwhelmingly through heterosexual contact. A model which gives age-specific prevalence rates in the presence of a progressing epidemic is fitted to prevalence data for 1998 using maximum likelihood methods and used to derive the age-specific incidence. Error estimates are obtained using a Monte Carlo procedure. Although the method is quite general some simplifying assumptions are made concerning the form of the risk function and sensitivity analyses are performed to explore the importance of these assumptions. The analysis shows that in 1998 the annual incidence of infection per susceptible woman increased from 5.4 per cent (3.3-8.5 per cent; here and elsewhere ranges give 95 per cent confidence limits) at age 15 years to 24.5 per cent (20.6-29.1 per cent) at age 22 years and declined to 1.3 per cent (0.5-2.9 per cent) at age 50 years; standardized to a uniform age distribution, the overall incidence per susceptible woman aged 15 to 59 was 11.4 per cent (10.0-13.1 per cent); per women in the population it was 8.4 per cent (7.3-9.5 per cent). Standardized to the age distribution of the female population the average incidence per woman was 9.6 per cent (8.4-11.0 per cent); standardized to the age distribution of women attending antenatal clinics, it was 11.3 per cent (9.8-13.3 per cent). The estimated incidence depends on the values used for the epidemic growth rate and the AIDS related mortality. To ensure that, for this population, errors in these two parameters change the age specific estimates of the annual incidence by less than the standard deviation of the estimates of the age specific incidence, the AIDS related mortality should be known to within +/-50 per cent and the epidemic growth rate to within +/-25 per cent, both of which conditions are met. In the absence of cohort studies to measure the incidence of HIV infection directly, useful estimates of the age-specific incidence can be obtained from cross sectional, age-specific prevalence data and repeat cross-sectional data on the overall prevalence of HIV infection. Several assumptions were made because of the lack of data but sensitivity analyses show that they are unlikely to affect the overall estimates significantly. These estimates are important in assessing the magnitude of the public health problem, for designing vaccine trials and for evaluating the impact of interventions. PMID- 11427957 TI - Multinomial analysis of smoothed HIV back-calculation models incorporating uncertainty in the AIDS incidence. AB - Back-calculation models, developed to reconstruct the past trend of human immunodeficiency virus (HIV) and to project future acquired immunodeficiency syndrome incidence (AIDS), are usually and unrealistically based on the assumption that the observed AIDS counts are independently distributed according to a Poisson process. In contrast, we argue that a multinomial framework is more suitable to this situation, leading to a natural covariance structure. The ill conditioned nature of the problem is solved by modelling the HIV parameters according to a cubic spline function to reduce the dimensionality of the parameter space and obtain smoother parameter estimates. We applied a regression spline technique which yields to a computationally stable basis incorporating the incubation period in the new design matrix. We directly incorporate the reporting delay distribution in the AIDS incidence data, leading to a more complex formulation of the variance and covariance model that is adapted to the iteratively reweighted least square (IRLS) algorithm. In this case we obtain more accurate estimates of the standard error of the HIV incidence, especially in the most recent time. Our model, which uses a cubic spline reparameterization based on a multinomial probability distribution, is applied to the AIDS epidemic data in Italy. PMID- 11427958 TI - The use of mixture models for identifying high risks in disease mapping. AB - Conventional approaches for estimating risks in disease mapping or mortality studies are based on Poisson inference. Frequently, overdispersion is present and this extra variability is modelled by introducing random effects. In this paper we compare two computationally simple approaches for incorporating random effects: one based on a non-parametric mixture model assuming that the population arises from a discrete mixture of Poisson distributions, and the second using a Poisson-normal mixture model which allows for spatial autocorrelation. The comparison is focused on how well each of these methods identify the regions which have high risks. Such identification is important because policy makers may wish to target regions associated with such extreme risks for financial assistance while epidemiologists may wish to target such regions for further study. The Poisson-normal mixture model is presented from both a frequentist, or empirical Bayes, and a fully Bayesian point of view. We compare results obtained with the parametric and non-parametric models specifically in terms of detecting extreme mortality risks, using infant mortality data of British Columbia, Canada, for the period 1981-1985, breast cancer data from Sardinia, for the period 1983 1987, and Scottish lip cancer data for 1975-1980. However, we also investigate the performance of these models in a simulation study. The key finding is that discrete mixture models seem to be able to locate regions which experience high risks; normal mixture models also work well in this regard, and perform substantially better when spatial autocorrelation is present. PMID- 11427959 TI - Isolation and sequence analysis of the gene encoding triose phosphate isomerase from Zygosaccharomyces bailii. AB - The ZbTPI1 gene encoding triose phosphate isomerase (TIM) was cloned from a Zygosaccharomyces bailii genomic library by complementation of the Saccharomyces cerevisiae tpi1 mutant strain. The nucleotide sequence of a 1.5 kb fragment showed an open reading frame (ORF) of 746 bp, encoding a protein of 248 amino acid residues. The deduced amino acid sequence shares a high degree of homology with TIMs from other yeast species, including some highly conserved regions. The analysis of the promoter sequence of the ZbTPI1 revealed the presence of putative motifs known to have regulatory functions in S. cerevisiae. The GenBank Accession No. of ZbTPI1 is AF325852. PMID- 11427960 TI - Disruption and phenotypic analysis of six open reading frames from chromosome VII of Saccharomyces cerevisiae reveals one essential gene. AB - Six open reading frames (ORFs) located on chromosome VII of Saccharomyces cerevisiae (YGR205w, YGR210c, YGR211w, YGR241c, YGR243w and YGR244c) were disrupted in two different genetic backgrounds using short-flanking homology (SFH) gene replacement. Sporulation and tetrad analysis showed that YGR211w, recently identified as the yeast ZPR1 gene, is an essential gene. The other five genes are non-essential, and no phenotypes could be associated to their inactivation. Two of these genes have recently been further characterized: YGR241c (YAP1802) encodes a yeast adaptor protein and YGR244c (LSC2) encodes the beta-subunit of the succinyl-CoA ligase. For each ORF, a replacement cassette with long flanking regions homologous to the target locus was cloned in pUG7, and the cognate wild-type gene was cloned in pRS416. PMID- 11427961 TI - A Saccharomyces servazzii clone homologous to Saccharomyces cerevisiae chromosome III spanning KAR4, ARS 304 and SPB1 lacks the recombination enhancer but contains an unknown ORF. AB - In order to learn about the evolutionary conservation of the recombination enhancer (RE) that controls donor preference during mating type switching in Saccharomyces cerevisiae, we have cloned a 13 kb region from S. servazzii. We find that the order of four genes surrounding the RE in S. cerevisiae (PRD1, KAR4, SPB1 and PBN1) is preserved in S.servazzii. However, there is an additional ORF in S. servazzii between PRD1 and KAR4 that is not homologous to any gene in S. cerevisiae or to genes in other organisms. Despite a 75-79% amino acid identity for KAR4 and SPB1, respectively, the S. servazzii sequence did not carry a well-conserved RE sequence and these sequences lacked RE function when introduced into S. cerevisiae. The S. servazzii region contains a sequence that supports autonomous DNA replication in S. cerevisiae and may represent a homologue of ARS304. The S. servazziii sequence has Genbank Accession No. BankIt359091 AF307954. PMID- 11427962 TI - High-level production of human type I collagen in the yeast Pichia pastoris. AB - Four human genes, two of them encoding the proalpha1 and proalpha2 chains of type I procollagen and two of them the two types of subunit of prolyl 4-hydroxylase (4 PH), were integrated into the genome of Pichia pastoris. The proalpha1 and proalpha2 chains expressed formed type I procollagen molecules with the correct 2:1 chain ratio, and the 4-PH subunits formed an active enzyme tetramer that fully hydroxylated the proalpha chains. Chains lacking their N but not C propeptides formed pCcollagen molecules with the 2:1 chain ratio and, surprisingly, the expression levels of pCcollagen were 1.5-3-fold relative to those of procollagen. Both types of molecule could be converted by pepsin treatment to collagen molecules that formed native-type fibrils in vitro. The expression levels obtained for the pCcollagen using only single copies of each of the four genes and a 2 l fermenter ranged up to 0.5 g/l, indicating that it should be possible to optimize this system for high-level production of recombinant human type I collagen for numerous medical applications. PMID- 11427963 TI - Gene order in a 10 275 bp fragment of Yarrowia lipolytica, including adjacent YlURA5 and YlSEC65 genes conserved in four yeast species. AB - We have determined the sequence of a 10275 bp DNA segment of Yarrowia lipolytica located on chromosome VI. The sequence contains six complete open reading frames (ORFs) longer than 100 amino acids and two more partial ORFs at both ends. Two of the ORFs encode for the well-characterized genes YlURA5 (orotate phosphoribosyltransferase) and YlSEC65 (encoding a subunit of the signal recognition particle). These two genes show an identical organization-located on opposite strands and in opposite orientations-in four yeast species: Saccharomyces cerevisiae, Kluyveromyces lactis, Candida albicans and Y. lipolytica. One ORF and the two partial ORFs code for putative proteins showing significant homology with proteins from other organisms. YlVI-108w (partial) and YlVI-103w show 39% and 54% identity, respectively, with YDR430c and YHR088w from S. cerevisiae. YlVI-102c (partial) shows significant homology with a matrix protein, lustrin A from Haliotis rufescens, and with the PGRS subfamily (Gly-rich proteins) of Mycobacterium tuberculosis. The three remaining ORFs show weak or non-significant homology with previously sequenced genes. The nucleotide sequence has been submitted to the EMBL database under Accession No. AI006754. PMID- 11427964 TI - Kluyveromyces lactis cytoplasmic plasmid pGKL2: heterologous expression of Orf3p and proof of guanylyltransferase and mRNA-triphosphatase activities. AB - The predicted ORF3 polypeptide (Orf3p) of the linear genetic element pGKL2 from Kluyveromyces lactis was expressed in Bacillus megaterium as a fusion protein with a His(6X)-tag at the C-terminus for isolation by Ni-affinity chromatography. This is the first time that a yeast cytoplasmic gene product has been expressed heterologously as a functional protein in a bacterial system. The purified protein was found to display both RNA 5'-triphosphatase and guanylyltransferase activities. When the lysine residue present at position 177 of the protein within the sequence motif (KXDG), highly conserved in capping enzymes and other nucleotidyl transferases, was substituted by alanine, the guanylyltransferase activity was lost, thereby proving an important role for the transfer of GMP from GTP to the 5'-diphosphate end of the mRNA. Our in vitro data provides the first direct evidence that the polypeptide encoded by ORF3 of the cytoplasmic yeast plasmid pGKL2 functions as a plasmid-specific capping enzyme. Since genes equivalent to ORF3 of pGKL2 have been identified in all autonomous cytoplasmic yeast DNA elements investigated so far, our findings are of general significance for these widely distributed yeast extranuclear genetic elements. PMID- 11427965 TI - Saccharomyces cerevisiae YCRO17c/CWH43 encodes a putative sensor/transporter protein upstream of the BCK2 branch of the PKC1-dependent cell wall integrity pathway. AB - The Saccharomyces cerevisiae cwh43-2 mutant, originally isolated for its Calcofluor white hypersensitivity, displays several cell wall defects similar to mutants in the PKC1-MPK1 pathway, including a growth defect and increased release of beta-1,6-glucan and beta-glucosylated proteins into the growth medium at increased temperatures. The cloning of CWH43 showed that it corresponds to YCR017c and encodes a protein with 14-16 transmembrane segments containing several putative phosphorylation and glycosylation sites. The N-terminal part of the amino acid sequence of Cwh43p shows 40% similarity with the mammalian FRAG1, a membrane protein that activates the fibroblast growth factor receptor of rat osteosarcoma (FGFR2-ROS) and with protein sequences of four uncharacterized ORFs from Caenorhabditis elegans and one from Drosophila melanogaster. The C-terminus of Cwh43p shows low similarities with a xylose permease of Bacillus megaterium and with putative sugar transporter from D. melanogaster, and has 52% similarity with a protein sequence from a Schizosaccharomyces pombe cDNA. A Cwh43-GFP fusion protein suggested a plasma membrane localization, although localization to the internal structure of the cells could not be excluded, and it concentrates to the bud tip of small budded cells and to the neck of dividing cells. Deletion of CWH43 resulted in cell wall defects less pronounced than those of the cwh43-2 mutant. This allele-specific phenotype appears to be due to a G-R substitution at position 57 in a highly conserved region of the protein. Genetic analysis places CWH43 upstream of the BCK2 branch of the PKC1 signalling pathway, since cwh43 mutations were synthetic lethal with pkc1 deletion, whereas the cwh43 defects could be rescued by overexpression of BCK2 and not by high-copy-number expression of genes encoding downstream proteins of the PKC1 pathway However, unlike BCK2, whose disruption in a cln3 mutant resulted in growth arrest in G(1), no growth defect was observed in a double cwh43 cln3 mutants. Taken together, it is proposed that CWH43 encodes a protein with putative sensor and transporter domains acting in parallel to the main PKC1-dependent cell wall integrity pathway, and that this gene has evolved into two distinct genes in higher eukaryotes. PMID- 11427966 TI - At acidic pH, the GPA2-cAMP pathway is necessary to counteract the ORD1-mediated repression of the hypoxic SRP1/TIR1 yeast gene. AB - The hypoxic SRP1/TIR1 gene encodes a stress-response cell wall mannoprotein and this gene is downregulated at acidic pH. The stress-responsive HOG pathway is necessary to maintain hypoxic TIR1 expression, but only at acidic pH. However, unlike known HOG pathway-dependent genes, TIR1 is under positive cAMP control and this effect is mediated by GPA2 but not by RAS2. Genetic analysis showed that ord1 mutation was epistatic to the gpa2 mutation, thereby indicating that Gpa2p is needed to counteract the Ord1 factor, which is involved in the repression of hypoxic TIR1 expression, while the HOG pathway appears to be independent from Ord1 repression. In addition, an increased ORD1 gene expression was observed in the Deltagpa2 mutant cells, meaning that GPA2 maintains a low basal level of ORD1 transcripts. Thus, cAMP allows partial relief of the TIR1 repression exerted by Ord1p. However, this is contradicted at acidic pH by the HOG pathway requirement because Hog1p is activated under stress conditions when the cAMP cellular content is low. The opposite effects of the GPA2-cAMP and HOG pathways are likely to explain the diminished hypoxic expression of TIR1 at acidic pH. PMID- 11427967 TI - A single-copy suppressor of the Saccharomyces cerevisae late-mitotic mutants cdc15 and dbf2 is encoded by the Candida albicans CDC14 gene. AB - The Saccharomyces cerevisiae CDC15, DBF2, TEM1 and CDC14 genes encode regulatory proteins that play a crucial role in the latest stages of the M phase of the cell cycle. By complementation of a S. cerevisiae cdc15-lyt1 mutant with a Candida albicans centromeric-based genomic library, we have isolated a homologue of the protein phosphatase-encoding gene CDC14. The sequence analysis of the C. albicans CDC14 gene reveals a putative open reading frame of 1626 base pairs interrupted by an intron located close to the 5' region. Analysis of C. albicans cDNA proved that the intron is processed in vivo. The CaCDC14 gene shares 49% of amino acid sequence identity with the S. cerevisiae CDC14 gene, 46% with Schizosaccharomyces pombe homologue, 35% with Caenorhabditis elegans and 37% and 38% with human CDC14A and CDC14B genes, respectively. As expected, the C. albicans CDC14 gene complemented a S. cerevisiae cdc14-1 mutant. We found that this gene was able to efficiently suppress not only a S.cerevisiae cdc15-lyt1 mutant but also a dbf2-2 mutant in a low number of copies and allowed growth, although very slightly, of a tem1 deletant. Overexpression of the human CDC14A and CDC14B genes complemented, although very poorly, S. cerevisiae cdc15-lyt1 and dbf2-2 mutants, suggesting a conserved function of these genes throughout phylogeny. The sequence of CaCDC14 was deposited in the EMBL database under Accession No. AJ243449. PMID- 11427968 TI - Cassettes for PCR-mediated construction of green, yellow, and cyan fluorescent protein fusions in Candida albicans. AB - We have developed a set of plasmids containing fluorescent protein cassettes for use in PCR-mediated gene tagging in Candida albicans. We engineered YFP and CFP variants of the GFP sequence optimized for C. albicans codon usage. The fluorescent protein sequences, linked to C. albicans auxotrophic marker sequences, were amplified by PCR and transformed directly into yeast. Gene specific sequence was incorporated into the PCR primers, such that the tag cassette integrates by homologous recombination at the 3'-end of the gene of interest. This technique was used to tag Cdc3 and Tub1 with GFP, YFP and CFP, which were readily visualized by fluorescence microscopy and localized as expected. In addition, Tub1-YFP and Cdc3-CFP were visualized in the same cells. Thus, this technique directs one-step construction of multiple fluorescent protein fusions, facilitating the study of protein co-expression and co localization in C. albicans cells in vivo. PMID- 11427969 TI - The diversity of retrotransposons in the yeast Cryptococcus neoformans. AB - We have undertaken an analysis of the retrotransposons in the medically important basidiomycetous fungus Cryptococcus neoformans. Using the data generated by a C. neoformans genome sequencing project at the Stanford Genome Technology Center, 15 distinct families of LTR retrotransposons and several families of non-LTR retrotransposons were identified. Members of at least seven families have transposed recently and are probably still active. For several families, only partial elements could be identified and these are quite diverse in sequence, suggesting that they are ancient components of the C. neoformans genome. Most C. neoformans elements are not closely related to previously identified fungal retrotransposons, suggesting that the diversity of fungal retrotransposons has been only sparsely sampled to date. C. neoformans has fewer distinct retrotransposon families than Candida albicans (37 or more), in particular fewer families represented solely by ancient and inactive elements, but it has considerably more families than either Saccharomyces cerevisiae (five) or Schizosaccharomyces pombe (two). The findings suggest that elimination of retrotransposons is faster in C. neoformans than in C. albicans, but perhaps not as rapid as in S. cerevisiae or Sz. pombe. The identification of the retrotransposons of C. neoformans should assist in the molecular characterization of this important pathogen, and also further our understanding of the role played by retroelements in genome evolution. PMID- 11427970 TI - Current awareness on yeast. AB - In order to keep subscribers up-to-date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly-published material on yeasts. Each bibliography is divided into 10 sections. 1 Books, Reviews & Symposia; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (4 weeks journals - search completed 4th Apr. 2001) PMID- 11427971 TI - PIs--do they really have all the answers? PMID- 11427972 TI - Bone Fos-silization. PMID- 11427973 TI - Eyeing-up stem cell transplantation. PMID- 11427974 TI - Idiopathic Down's syndrome cluster in Ireland. PMID- 11427975 TI - Lasers aimed at mad cows. PMID- 11427976 TI - Scaffolding in pigs offers cardiovascular gene therapy. PMID- 11427977 TI - Cannibalism reveals vCJD-susceptibility. PMID- 11427978 TI - Split-personality RANTES mutation. PMID- 11427979 TI - Gene therapy for the head and neck. PMID- 11427980 TI - Kissing and Kaposi's sarcoma. PMID- 11427981 TI - Black Death. PMID- 11427982 TI - Molecular genetics of pseudoxanthoma elasticum: a metabolic disorder at the environment-genome interface? AB - Pseudoxanthoma elasticum (PXE) is a relatively rare heritable disorder affecting the skin, eyes and cardiovascular system, with considerable morbidity and mortality. The disease affects the elastic fibers of affected organs, which become progressively calcified. Thus, PXE has been considered as a prototypic heritable connective tissue disorder affecting the elastic fiber system. Recently, PXE has been linked to mutations in the MRP6/ABCC6 gene, a member of the ABC transporter family, expressed primarily in the liver and the kidneys. This information, together with clinical observations suggesting environmental, hormonal and/or dietary modulation of the disease, raises the intriguing possibility that PXE is a primary metabolic disorder at the environment-genome interface. PMID- 11427983 TI - Molecular regulation of lymphangiogenesis and targets for tissue oedema. AB - New insight has recently been obtained into the molecular mechanisms regulating the function of lymphatic endothelial cells. Vascular endothelial growth factors C and -D have been shown to stimulate lymphangiogenesis, and their receptor VEGFR 3 has been linked to human hereditary lymphoedema, although there is evidence that other genes are also involved. These data suggest that it may become possible to stimulate lymphatic growth and function and to treat tissue oedema involved in many diseases. PMID- 11427984 TI - BSE crisis--transmission through blood transfusions? PMID- 11427985 TI - Preimplantation genetic diagnosis: applications for molecular medicine. AB - Preimplantation genetic diagnosis is an alternative to prenatal diagnosis for the detection of genetic disorders. Tests are conducted on single cells biopsied from embryos before they are implanted, allowing the selection of unaffected embryos before a pregnancy has been established. Thus, the issue of pregnancy termination is circumvented. The use of preimplantation genetic diagnosis might have a significant impact on in vitro fertilization success rates as well as allowing the diagnosis of inherited disease. PMID- 11427986 TI - Cord blood: from waste product to valuable resource. PMID- 11427987 TI - Novel clinical strategies for the treatment of pancreatic carcinoma. AB - Pancreatic carcinoma ranks as the eighth most frequent type of solid tumour arising worldwide yet it represents the fourth most frequent cause of death. This discrepancy reflects the current lack of effective treatment available for the pancreatic cancer patient and highlights the urgent need for new therapeutic principles in this area. The last five years have seen an increasing number of novel approaches both in the pre-clinical area as well as in clinical trials for pancreatic cancer treatments. This review summarizes these new developments and attempts to rationalize the possibilities available for the patient at the beginning of the new millennium. PMID- 11427988 TI - Disease model: LAMP-2 enlightens Danon disease. AB - Danon disease ('lysosomal glycogen storage disease with normal acid maltase') is characterized by a cardiomyopathy, myopathy and variable mental retardation. Mutations in the coding sequence of the lysosomal-associated membrane protein 2 (LAMP-2) were shown to cause a LAMP-2 deficiency in patients with Danon disease. LAMP-2 deficient mice manifest a similar vacuolar cardioskeletal myopathy. In addition to the patient reports LAMP-2 deficiency in mice causes pancreatic, hepatocytic, endothelial and leucocyte vacuolation. LAMP-2 deficient mice represent a valuable animal model of Danon disease. They will further be used to study the exact role of LAMP-2 in autophagy and to analyse the consequences of an impaired autophagic pathway in various tissues. PMID- 11427989 TI - UK genetic testing and insurance decision provokes mixed response. PMID- 11427990 TI - Extending the ethical boundaries of stem cell research. PMID- 11427991 TI - Genes and brains, molecular medicine and neuropathology. PMID- 11427992 TI - Too much hypothalamic serotonin transporter is bad for your mood. PMID- 11427993 TI - Pseudohypoparathyroidism: new insight into Gs alpha. PMID- 11427994 TI - Protein shuttles, IGF-I and colorectal cancer. PMID- 11427995 TI - Benefits from early treatment of HIV-1 infection. PMID- 11427996 TI - Functional properties of failing human ventricular myocytes. AB - Reduced peak systolic Ca2+ and slow decay of the Ca2+ transient are common features of the end-stage failing human ventricular myocyte and are thought to underlie abnormal ventricular contractility in congestive heart failure (CHF). Individual changes in the expression or activity of Ca2+ transport proteins of the sarcoplasmic reticulum (SR Ca2+ ATPase, SERCa) or the sarcolemmal (sodium calcium exchanger, NCX) have not always been observed in CHF and cannot per se consistently explain these Ca2+ transient defects. We review recent data that suggests that the normal balance of transport activities of SERCa and NCX is deranged in failing human myocytes. We hypothesize that an increase in the NCX/SERCa transport capacity in failing myocytes can explain the abnormal Ca2+ homeostasis of the failing human ventricular myocyte. PMID- 11427997 TI - The biology of myocardial hibernation. AB - Patients with chronic coronary artery disease frequently have contractile dysfunction that recovers upon reperfusion. The concept of myocardial hibernation views the observed reduction in contractile function not as the result of an ongoing energetic deficit, but as an adaptive down-regulation that serves to maintain myocardial integrity and viability. In the experiment, sustained perfusion-contraction matching, recovery of energy and substrate metabolism during ongoing ischemia, the potential for recruitment of inotropic reserve, lack of necrosis, and therefore recovery of function upon reperfusion are established features of hibernation. Apart from reduced calcium responsiveness, the underlying mechanisms are still unclear. In patients, the importance of reduced baseline blood flow vs. that of superimposed repetitive stunning is somewhat controversial; however, in most studies blood flow is reduced, and the myocardium must be ischemic often enough to have persistent dysfunction. Morphologically, hibernating myocardium displays features of dedifferentiation, with loss of cardiomyocytes and myofibrils, and of degeneration, with increased interstitial fibrosis. Patients with hibernating myocardium must be identified and undergo revascularization. With a better understanding of the underlying mechanisms of hibernation, these adaptive responses to ischemia can potentially be recruited and reinforced pharmacologically to delay impending myocardial infarction. PMID- 11427998 TI - Molecular and ionic basis of congenital complete heart block. AB - Congenital heart block (CHB), detected at or before birth in a structurally normal heart, is strongly associated with autoantibodies reactive with the intracellular soluble ribonucleoproteins 48kD SSB/La, 52kD SSA/Ro, and 60kD SSA/Ro. CHB is presumed to be due to the transplacental passage of autoantibodies from the mother into the fetal circulation. Varying degrees of heart block have been reported. Although second degree block has, on rare occasion, reverted to normal sinus rhythm, complete atrio-ventricular (AV) block is irreversible. CHB carries substantial mortality and morbidity, with > 60% of affected children requiring lifelong pacemakers. The recurrence rate exceeds, by at least twofold, that of the first birth and is likely to influence the decision to have more children. Curiously, the mother's heart is almost never affected (with complete heart block) despite exposure to identical circulating autoantibodies. As part of our continuing effort to understand the complex factors contributing to the pathogenesis of CHB, we have established an animal model of CHB by immunizing female mice with recombinant proteins/antigens, reproduced the human complete AV block in an isolated Langendorff perfused fetal heart, and correlated these findings with L-type Ca channel inhibition by maternal antibodies from mothers of children with CHB. In addition, we established a passive animal model by directly injecting maternal antibodies into pregnant mice and reported significant sinus bradycardia, indicating that the spectrum of conduction abnormalities may extend beyond the AV node. All together, the data provided strong evidence supporting an etiologic role of antibody/Ca channel involvement in the pathogenesis of CHB. However, other yet unknown factors seem necessary to explain the full expression of CHB. PMID- 11427999 TI - Invasive cardiac electrophysiology in the mouse: techniques and applications. AB - As the genetic nature of a wide spectrum of cardiovascular diseases is being elucidated, it is increasingly important to understand the functional role of specific genes on cardiac arrhythmia and conduction disturbances. The progress made in molecular genetics has allowed the creation of mice with targeted gene overexpression or elimination. These animals are valuable tools for researchers who have adapted their clinical and technical skills to the mouse, in order to extract information on the phenotypic consequences of the specific genetic disruption. In this review, we summarize the progress made in the field of invasive murine electrophysiology, focusing on the recent technical advances in in vivo electrophysiologic testing and its application to various genetically engineered mouse models. The authors' views on the future needs and trends in the field are also presented. PMID- 11428000 TI - TGF-beta in the cardiovascular system: molecular mechanisms of a context-specific growth factor. AB - Transforming growth factor beta-1 is the prototypical member of a class of growth factors whose actions have been strongly implicated in a number of pathophysiologic processes including chronic vascular diseases such as atherosclerosis and hypertension. One of the hall-marks of this class of growth factors is the diverse nature of their actions; a characteristic that is thought to arise from the fact that the effects of these factors are very dependent upon the particular cellular context in which they operate. There has been substantial progress in understanding the molecular signaling mechanisms utilized by these factors. These findings are beginning to provide a mechanistic framework with which to understand the complex and pleiotropic actions of these factors on cells and tissues of the cardiovascular system. PMID- 11428001 TI - Atrial form and function: lessons from human molecular genetics. AB - Molecular genetic analyses of human hereditary disorders that affect cardiac atrial structure and function have recently identified several genes that regulate atrial morphogenesis. Mutations of the TBX5, NKX2.5, EVC, and PRKAR1 alpha genes all result in abnormalities of human atrial growth and development, and mutations in at least one gene results in familial atrial fibrillation and is as yet unidentified. Ongoing studies to find interactions between these transcription factors and intracellular signaling molecules and other as yet unknown genes are establishing critical pathways in human cardiogenesis. Human investigation and experimental animal models of heart development synergize to elucidate etiologies of common congenital heart disease. PMID- 11428002 TI - Pd-catalyzed asymmetric hydrogenation of alpha-fluorinated iminoesters in fluorinated alcohol: a new and catalytic enantioselective synthesis of fluoro alpha-amino acid derivatives. AB - [figure: see text] Under hydrogen pressure, a catalytic amount of palladium(II) trifluoroacetate and 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP) promoted asymmetric hydrogenation of alpha-fluorinated iminoesters to afford highly enantioenriched beta-fluorinated alpha-amino esters. The yield and ee were much improved by employing fluorinated alcohols such as 2,2,2-trifluoroethanol (up to 91% ee). PMID- 11428003 TI - A versatile, three-component-reaction route to N-glycosylamines. AB - [figure: see text] Under the agency of N-bromosuccinimide, n-pentenyl glycosides, acetonitrile, and carboxylic acids participate in three-component-reactions that afford N-acylated glycosylamines. The procedure tolerate diverse donors, and C2 tetrachlorophthalimido and C2-azido groups effectively control anomeric stereoselectivity. Success of the procedure does not appear to depend on the acid's strength, but for an aromatic acid, substitution pattern affects the rate, while the presence of a lone pair on the para substituent inhibits the process. PMID- 11428004 TI - Synthesis of ethyl 5-O-(alpha-D-arabinofuranosyl)-6-O-(beta-D-galactofuranosyl) beta-D- galactofuranoside present in motif E of the Mycobacterium tuberculosis cell wall. AB - [figure: see text] The stereocontrolled synthesis of the trisaccharide ethyl 5-O (alpha-D-arabinofuranosyl)-6-O-(beta-D-galactofuranosyl)- beta-D galactofuranoside present in motif E of the Mycobacterium tuberculosis cell wall is described. PMID- 11428005 TI - A first comparison of solid state and solution structure of a complex between lanthanum nitrate and 1,9-diaza-18-crown-6. AB - [figure: see text] The structure of the complex between lanthanum(III)nitrate and 1,9-diaza-18-crown-6 is analyzed with three independent methods. The conformation observed by analysis of vicinal NMR coupling constants in solution agrees with both the one found by a single crystal X-ray study and with simulations by molecular orbital and force field calculations. In the acetonitrile solution, the NMR data show a bent crown conformation with nearly C2v symmetry, which is stable on the NMR time scale. PMID- 11428006 TI - In search of catalytically active species in the surfactant-mediated biphasic alkene epoxidation with Mimoun-type complexes. AB - [figure: see text] A biphasic protocol for the catalytic olefin epoxidation with Mimoun-type complexes [MoO(O2)2(OPR3)] (1) was recently patented by BASF. Density functional calculations have been carried out to identify potentially active species in addition to the parent complex 1. It has been found that the (mu 2,eta 1:eta 2-O2)-bridged dimer [MoO(O2)2(OPR3)]2 is significantly less reactive than the monomer. The calculations show that the parent complex is strongly activated by protons coordinating with the peroxo functionalities. PMID- 11428007 TI - Synthesis of a fully functionalized protected C1-C11 fragment for the synthesis of the tedanolides. AB - [figure: see text] The use of several non-aldol aldol processes allows one to prepare a fully functionalized and completely protected C1-C11 fragment that should be useful for the total synthesis of the tedanolides. PMID- 11428008 TI - Solid-phase synthesis of beta-lactams via the Miller hydroxamate approach. AB - [figure: see text] beta-Lactams were prepared on solid phase starting from serine, threonine, or other beta-hydroxyacids derived from naturally occurring amino acids and a resin bound hydroxylamine. The ring closure was carried out under Mitsunobu conditions. The amino group present on the beta-lactam was used to assemble a short peptide. After a reductive cleavage with Sml2, beta-lactam containing peptides were obtained. PMID- 11428009 TI - Asymmetric synthesis of alpha,beta-dialkyl-alpha-phenylalanines via direct alkylation of a chiral alanine derivative with racemic alpha-alkylbenzyl bromides. A case of high enantiomer differentiation at room temperature. AB - [figure: see text] This study demonstrates that the direct alkylation of a Ni(II) complex of the chiral Schiff base of alanine with (S)-o-[N-(N-benzylprolyl)amino] benzophenone, with racemic alpha-alkylbenzyl bromides, is a synthetically feasible and methodologically advantageous approach to the target alpha,beta dialkylphenylalanines over previously reported methods. For the first time we report and rationalize a case of a high enantiomer differentiation process at room temperature. PMID- 11428010 TI - Stereocontrolled total synthesis of alkaloid G via the oxy-anion Cope rearrangement and improved total synthesis of (+)-ajmaline. AB - [figure: see text] The oxy-anion Cope rearrangement followed by protonation of the enolate which resulted under conditions of kinetic control has been employed to generate the key asymmetric centers at C(15), C(16), and C(20) in alkaloid G (1) and (+)-ajmaline (2) in a highly stereocontrolled fashion. The aldehyde 7b from this process has been converted into alkaloid G (1) and (+)-ajmaline (2) in 36% and 13% overall yields (11 reaction vessels from 3), respectively. PMID- 11428011 TI - Iodination of organic substrates with halide salts and H2O2 using an organotelluride catalyst. AB - [figure: see text] Organotelluride 1 is a water-soluble catalyst for the oxidation of iodide with hydrogen peroxide in pH 6 phosphate buffer. In two-phase systems, organic substrates are efficiently iodinated using 0.8 mol % of catalyst. Water-soluble substrates are iodinated without an organic cosolvent. PMID- 11428012 TI - Exploiting the Curtin-Hammett principle--recognition-mediated acceleration of an aldol reaction. AB - [figure: see text] Curtin-Hammett-Winstein-Holness kinetics are exploited in the acceleration of the aldol reaction between formylbenzo-15-crown-5 and acetylbenzo 15-crown-5. Potassium cations facilitate this acceleration through the fast, reversible formation of a 1:1:1 sandwich complex between the crown ethers and the metal cation. PMID- 11428013 TI - Connectivity effects in isomeric naphthalenedinitrenes. AB - [figure: see text] UV-vis, FTIR, and ESR spectroscopic studies were carried out on samples of 1,4- and 1,5-naphthalenediazide that were photolyzed at cryogenic temperatures in argon and frozen solvent matrices. Mononitrenes and diiminediyl systems are produced. Spectral and computational results are consistent with quinonoidal singlet ground-state structures for the diiminediyls, rather than aromatic dinitrene structures. PMID- 11428014 TI - Organolanthanide-catalyzed cyclization/boration of 1,5- and 1,6-Dienes. AB - [figure: see text] 1,5- and 1,6-Dienes undergo a cyclization/boration reaction in the presence of a catalytic amount of Cp*2Sm.THF. The resulting organoboranes can be oxidized to the corresponding primary cyclic alcohols using standard conditions. PMID- 11428015 TI - Synthesis of 3-indolyl-2,5-dihydroxybenzoquinones. AB - [figure: see text] 3-Indolylquinones can be efficiently prepared by the acid catalyzed condensation of Indoles with 2,5-dichiorobenzoquinone, followed by DDQ oxidation. The resulting dichloroquinones are hydrolyzed to the 3 indolyidihydroxybenzoquinones. The 3-indolylquinone substructure is of interest because of its presence in natural products that modulate biological processes through protein-protein interactions, including the asterriquinones. PMID- 11428016 TI - Synthesis of cyclic sulfonamides through intramolecular Diels-Alder reactions. AB - [figure: see text] Substituted 2,3,3a,4,5,7a-hexahydrobenzo[d]isothiazole 1,1 dioxides and 3,4,4a,5,6,8a-hexahydro-2H-benzo[e][1,2]thiazine 1,1-dioxides, novel cyclic sulfonamides, were synthesized by the thermal Diels-Alder reaction of triene derivatives of buta-1,3-diene-1-sulfonic acid amide. The stereochemical outcome of the reaction was determined by NMR spectroscopy and X-ray crystallographic analysis. This chemistry has been used for the synthesis of 2-(4 chlorobenzyl)-5-(1H-imidazol-4-yl)-2,3,3a,4,5,7a-hexahydrobenzo[d] isothiazole 1,1-dioxide, a histamine H3 receptor antagonist. PMID- 11428017 TI - Highly enantioselective hydrogenation of alpha-dehydroamino acids by rhodium complexes with new unsymmetric P-chirogenic bisphosphine ligands. AB - [figure: see text] New rhodium catalysts with unsymmetric P-chirogenic bis(phosphino)ethanes, BisP*-Rh, exhibited very high enantioselectivity (98-99%) in the hydrogenation of alpha-dehydroamino acid derivatives. Such high enantioselectivity should result from the asymmetric environment around the Rh atom, as was shown in the molecular structure of the catalyst analyzed by X-rays. The asymmetry can be controlled by the combination of the alkyl groups on the two phosphorus atoms. PMID- 11428018 TI - Stereoselective synthesis of 2-S-phenyl-2-deoxy-beta-glycosides using phenyl 2,3 O-thionocarbonyl-1-thioglycoside donors via 1,2-Migration and concurrent glycosidation. AB - [figure: see text] 1,2-Migration and concurrent glycosidation of phenyl 2,3-O thionocarbonyl-1-thlo-alpha-L-rhamnopyranosides under the action of methyl trifluoromethanesulfonate (MeOTf) afforded in high yields the 3-O (methylthio)carbonyl-2-S-phenyl-2,6-dideoxy-beta-L-glucopyranosides, ready precursors to the corresponding 2-deoxy-beta-glycosides. PMID- 11428019 TI - Photo amidoglycosylation of an allal azidoformate. Synthesis of beta-2-amido allopyranosides. AB - [figure: see text] Photolysis of an allal C-3 azidoformate provoked intramolecular nitrene insertion into the glycal C=C unit and allowed direct incorporation of alcohol nucleophiles as beta-disposed substituents at C-1. The 2 amido allopyranoside products were elaborated via N-acylation and selective oxazolidinone hydrolysis, providing N-Boc-protected 2-amino sugars and simplifying stereochemical assignments. Synthesis of the potentially labile allal azidoformate was achieved via reaction of the corresponding carbonyl imidazolide with trimethylsilyl azide, facilitated by dibutyltin oxide. PMID- 11428020 TI - Cyclization/hydrosilylation of functionalized diynes catalyzed by a cationic platinum phenanthroline complex. AB - [figure: see text] A 1:1 mixture of the platinum phenanthroline complex (phen)PtMe2 and B(C6F5)3 catalyzed the cyclization/hydrosilylation of functionalized 1,6- and 1,7-diynes to form silylated 1,2-dialkylidenecycloalkanes in good yield and with high Z-selectivity. PMID- 11428021 TI - A new polymer-anchored chiral catalyst for asymmetric Michael addition reactions. AB - [figure: see text] Monomer (R,R)-3-aza-3-(p-vinylbenzyl)-1,5-diphenyl-1,5 dihydroxypentane (2) when polymerized with styrene and divinylbenzene affords polymers, onto which lithium and aluminum are incorporated via reaction with lithium aluminum hydride. The resulting insoluble polymers containing chiral lithium and aluminum active centers are quite effective for asymmetric Michael addition of nitro compounds, thiols, and amines. The optimized reaction conditions yield Michael adducts in good yield with high enantiomeric excesses. PMID- 11428022 TI - Cross-coupling reactions of potassium alkyltrifluoroborates with aryl and 1 alkenyl trifluoromethanesulfonates. AB - [figure: see text] The palladium-catalyzed coupling reaction of potassium alkyltrifluoroborates with aryl- or alkenyltriflates proceeds to afford the corresponding arenes or alkenes in high yield. The borates are all solids, stable in air, and thus can be stored on the shelf indefinitely. The cross coupling can be effected using PdCl2(dppf).CH2Cl2 as the catalyst in THF-H2O in the presence of Cs2CO3. A variety of functional groups can be tolerated within the borate and/or the triflate coupling partner. PMID- 11428023 TI - Carbon hydroxylation of alkyltetrahydropyranols: a paradigm for spiroacetal biosynthesis in Bactrocera sp. AB - [figure: see text] In a number of Bactrocera species the penultimate step in the biosynthesis of spiroacetals is shown to be the hydroxylation of an alkyltetrahydropyranol followed by cyclization. The monooxygenases that catalyze this side chain hydroxylation show a strong preference for oxidation four carbons from the hemiketal center, to produce the spiroacetal. The hydroxy spiroacetals observed in Bactrocera appear to derive from direct oxidation of the parent spiroacetals and not from alternate precursors. PMID- 11428024 TI - Synthesis of D- and L-deoxymannojirimycin via an asymmetric aminohydroxylation of vinylfuran. AB - [figure: see text] The Sharpless catalytic asymmetric aminohydroxylation has been applied to 2-vinylfuran, producing beta-hydroxyfurfurylamine 5a with enantioexcess of > 86% and 21% yield from furfural. The Cbz and TBS protected amino alcohol 5a was converted into both the D- and L-isomers of deoxymannojirimycin (DMJ) and deoxygulonojirimycin in five to seven steps and 48% and 66% overall yields. The key steps include the use of an aza-Achmatowicz reaction, a diastereoselective Luche reduction, diastereoselective dihydroxylation, and a tandem Cbz deprotection/reductive amination. PMID- 11428025 TI - Synthesis of S-linked glycosyl amino acids in aqueous solution with unprotected carbohydrates. AB - [figure: see text] The cyclic sulfamidate 5 was synthesized in 60% overall yield from L-serine benzyl ester. Compound 5 reacted cleanly with the sodium thiolate salt of a variety of unprotected 1-thio sugars in aqueous buffer to afford the corresponding S-linked amino acid glycoconjugates in good yields after hydrolysis of the N-sulfates. PMID- 11428026 TI - Proton-assisted switching of reaction pathways of stilbene analogues brought by direct irradiation. AB - [figure: see text] Photochemical reaction pathways during direct irradiation of stilbene analogues (1a-1d) can be switched completely by adjusting the concentration of hydrochloric acids. Competitive ring opening and acid-catalyzed hydrolysis processes are responsible for this novel selectivity. PMID- 11428027 TI - Transannular nitrone cycloaddition. A stereocontrolled entry to the spirocyclic core of pinnaic acid. AB - [figure: see text] Thermolysis of lactone 18 initiated a stereospecific transannular nitrone-olefin [3 + 2] cycloaddition to yield tetracycle 19. Methanolysis followed by reductive cleavage of the isoxazolidine yielded 20, representing the azaspirocyclic core of pinnaic acid (1). PMID- 11428028 TI - A "hard/soft" mismatch enables catalytic Friedel-Crafts acylations. AB - [figure: see text] Cationic complexes of Pt(II) and other late transition metals efficiently catalyze Friedel-Crafts acylations of moderately activated arenes by carboxylic acid anhydrides. The nature of the catalytically relevant species formed from (PhCN)2PtCl2 and AgSbF6 and their interactions with the substrates are studied by NMR and ESI-MS. PMID- 11428029 TI - Rhodium complex-catalyzed reaction of isonitriles with carbonyl compounds: catalytic synthesis of pyrroles. AB - [figure: see text] Low-valent rhodium complexes are efficient catalysts for the activation of alpha-C-H bond of isonitriles. Addition of isonitriles to carbonyl compounds proceeds under mild and neutral conditions to give the corresponding alpha,beta-unsaturated formamides. Catalytic synthesis of pyrroles can be performed by cyclocondensation of isonitriles with 1,3-dicarbonyl compounds. PMID- 11428030 TI - On the inherent instability of alpha-amino alpha'-fluoro ketones. Evidence for their transformation to reactive oxyvinyliminium ion intermediates. AB - [figure: see text] alpha-Amino alpha'-fluoro ketones are shown to be inherently unstable intermediates. Evidence is presented that they undergo enolization toward the amino group followed by expulsion of fluoride ion, forming a proposed oxyvinyliminium ion (amino-substituted oxyallyl cation). In protic, nucleophilic media the proposed intermediate is trapped by solvent. In the presence of a reactive diene, [4 + 3] cycloadducts have been isolated. Prior observations concerning fluorinated amino ketones are discussed in light of these findings. PMID- 11428031 TI - First total synthesis of mosin B. AB - [figure: see text] The first total synthesis of mosin B and a diastereomer was accomplished using asymmetric desymmetrization of the sigma-symmetric diol and the Nozaki-Hiyama-Kishi reaction as the key steps. The THF core segment was stereoselectively constructed employing a stereodivergent synthesis starting from a common intermediate, 4-cyclohexene-1,2-diol, based on a desymmetrization strategy. By virtue of these synthetic results, it is suggested that the absolute configuration is 1a. PMID- 11428032 TI - Unprecedented migration of N-alkoxycarbonyl groups in protected pyroglutaminol. AB - [figure: see text] Cleavage of an O-silyl ether in an N-BOC-protected pyroglutaminol using TBAF led to an unprecedented migration of the BOC group. An investigation of the mechanism, based on experimental data and quantum mechanical calculations, is presented. Similar migration was observed for N-Cbz and N methoxycarbonyl groups. PMID- 11428033 TI - Bogorol A produced in culture by a marine Bacillus sp. reveals a novel template for cationic peptide antibiotics. AB - [figure: see text] Bogorol A (1), a novel peptide antibiotic active against MRSA and VRE, has been isolated from cultures of a marine Bacillus sp. collected in Papua New Guinea. The structure of bogorol A was elucidated by a combination of spectroscopic analyses and chemical degradation. Bogorol A illustrates a new structural template for "cationic peptide antibiotics". PMID- 11428034 TI - Functionalized bicyclo[3.3.0]octanes by enantioselective transannular desymmetrization. AB - [figure: see text] Enantioselective alpha-deprotonation-rearrangement of achiral substituted cyclooctene oxides 7, 17, and 18 using organolithiums in the presence of (-)-sparteine (3) or (-)-alpha-isosparteine (4) gives the functionalized bicyclo[3.3.0]octan-2-ols 8, 19, and 20 in 56-72% yields and 83-89% ee's. PMID- 11428035 TI - Facile oxidation of fused 1,4-dimethoxybenzenes to 1,4-quinones using NBS: fine tuned control over bromination and oxidation reactions. AB - [figure: see text] Fused 1,4-dimethoxybenzenes could be oxidized to benzoquinones by either direct oxidation or demethylation-oxidation. The oxidative demethylation of 5,8-dimethoxy-2-methylquinoline using 1.1 equiv of NBS in aqueous THF and a catalytic amount of H2SO4 at 20 degrees C for 5 min gave 2 methylquinoline-5,8-dione in 98% yield without bromination. Moreover, we can control either bromination or oxidative demethylation, or both reactions. PMID- 11428036 TI - Exploiting the reversibility of olefin metathesis. Syntheses of macrocyclic trisubstituted alkenes and (R,R)-(-)-pyrenophorin. AB - [figure: see text] The formation of the trisubstituted cycloalkene 7 by RCM of diene 5 proceeds via the acyclic dimer 6, thus demonstrating the ready reversibility of olefin metathesis if catalyzed by "second generation" ruthenium carbene complexes such as 2-4. When applied to acrylate 11, these catalysts trigger a cyclooligomerization process that evolves with time and serves as key step en route to the lactide antibiotic (-)-pyrenophorin 8. PMID- 11428037 TI - Studies on the total synthesis of formamicin: synthesis of the C(1)-C(11) fragment. AB - [figure: see text] An efficient and highly concise synthesis of 6, corresponding to the C(1)-C(11) fragment of formamicin (1), has been accomplished by a route utilizing a diastereoselective lactate aldol reaction to set the C(6) tertiary ether and the TES-OTf mediated transketalization of the C(6) tertiary methoxymethyl ether and the C(25) PMB ether to set the seven-membered methylene acetal unit (see 37-->38). PMID- 11428038 TI - Enantioselective rhodium(I)-catalyzed hydrogenation of trifluoromethyl ketones. AB - [figure: see text] The asymmetric hydrogenation of trifluoromethyl ketones to yield chiral alpha-trifluoromethyl alcohols with enantiomeric excesses up to 98% was achieved in the presence of chiral rhodium-(amidephosphine-phosphinite) complexes. PMID- 11428039 TI - First direct deprotonation-electrophile trapping of simple epoxides: synthesis of alpha,beta-epoxysilanes from terminal epoxides. AB - [figure: see text] The presence of a suitable diamine ligand is the key to achieving ring lithlation-substitution of epoxides without the need for activating substituents on the epoxide and constitutes a new synthetic entry to trans-alpha,beta-epoxysilanes. PMID- 11428040 TI - Palladium and Raney nickel catalyzed methanolic cleavage of stable borane-amine complexes. AB - [figure: see text] Palladium and Raney nickel were found to catalyze the methanolysis of borane-amine adducts. Hence, strongly complexed amines can now be liberated by simple treatment with Pd/C or Raney Ni in methanol. The method is applicable to primary, secondary, tertiary, and aromatic amines, and the mildness of the reaction conditions allows preservation of otherwise labile functional groups. PMID- 11428041 TI - Enantiopure 2,3-dihydro-4-pyridones as synthetic intermediates: a concise asymmetric synthesis of (+)-allopumiliotoxin 267A. AB - [figure: see text] A concise asymmetric synthesis of (+)-allopumillotoxin 267A has been accomplished using an enantiopure dihydropyridone building block. The synthesis is highly stereoselective and requires 10 steps from readily available material. PMID- 11428042 TI - A concise enantioselective synthesis of a key A-ring synthon for 1 alpha hydroxyvitamin D3 compounds. AB - [figure: see text] This report describes a concise enantioselective synthesis of the A-ring synthon for the synthesis of 1 alpha-hydroxyvitamin D3 compounds. The synthesis involves two notable transformations: (I) stereoselective construction of the enol triflate from the vinyl ketone by Michael addition of Ph2P(O)Li followed by in situ triflation of the resulting enolate and (II) palladium catalyzed Heck type cyclization of the enol triflate. PMID- 11428043 TI - Lewis acid-catalyzed ring-opening reactions of semicyclic N,O-acetals. AB - [figure: see text] Ring-opening reactions of semicyclic N,O-acetals with various nucleophiles such as silyl enol ethers are effectively catalyzed by a Lewis acid (TMSOTf). Reactions of 3-substituted N,O-acetals showed high diastereoselectivities. Synthetic utility of this method has been demonstrated in the stereoselective synthesis of an anti-malarial agent, isofebrifugine. PMID- 11428044 TI - Oxabicyclo[3.2.1]octenes in organic synthesis--direct ring opening of oxabicyclo[3.2.1] systems employing silyl ketene acetals in concentrated solutions of lithium perchlorate-diethyl ether: application to the synthesis of the C(19)-C(27) fragment of rifamycin S. AB - [figure: see text] The direct opening at the bridgehead of oxabicyclo[3.2.1]octenes employing silyl ketene acetals in 4.0-5.0 M lithium perchlorate in diethyl ether has been realized, which gives rise to highly functionalized cycloheptadienes that can be further manipulated for use in natural product synthesis. The bridgehead opening reaction has been employed in the construction of the C(19)-C(27) fragment of Rifamycin S. PMID- 11428045 TI - Synthesis of pyrrolidine C-nucleosides via Heck reaction. AB - [figure: see text] A novel method for the synthesis of pyrrolidine C-nucleosides has been developed. The key step of the synthesis is the palladium(0)-mediated coupling of a disubstituted N-protected 2-pyrroline and 5-iodouracil. C Nucleoside 14 and its N-methyl derivative 15 can easily be converted to the corresponding phosphoramidite building blocks for DNA synthesis. PMID- 11428046 TI - Identification of cytoskeletal markers for the different microvilli and cell types of the rat vomeronasal sensory epithelium. AB - The vomeronasal organ (VNO) of the mammal nose is specialized to detect pheromones. The presumed site of the chemosensory signal transduction of pheromones is the vomeronasal brush border of the VNO sensory epithelium, which has been shown to contain two different sets of microvilli: (i) the tall microvilli of supporting cells and (ii) the short microvilli of the chemoreceptive VNO neurons that branch and intermingle with the basal portions of the longer supporting cell microvilli. A key problem when studying the subcellular distribution of possible VNO signal transduction molecules at the light microscope level is the clear discrimination of immunosignals derived from dendritic microvilli of the VNO neurons and surrounding supporting cell structures. In the present study we therefore looked for cytoskeletal marker proteins, that might help to distinguish at the light microscope level between the two sets of microvilli. By immunostaining we found that the VNO dendritic microvilli can be selectively labelled with antibodies to the calcium-sensitive actin filament-bundling protein villin, whereas supporting cell microvilli contain the actin filament cross-linking protein fimbrin, but not villin. Useful cytoplasmic marker molecules for cellular discrimination were cytokeratin 18 for supporting cells and beta-tubulin for dendrites of VNO neurons. A further finding was that the non-sensory epithelium of the rat VNO contains brush cells, a cell type that appears to be involved in certain aspects of chemoreception in the gut. Brush cells or other structures of the vomeronasal brush border did not contain alpha-gustducin. PMID- 11428047 TI - Astrocyte invasion and vasculogenesis in the developing ferret retina. AB - We studied the time course of astrocyte invasion and blood vessel formation in the developing ferret retina using glial fibrillary acidic protein (GFAP) immunohistochemistry for astrocytes and isolectin B4 histochemistry for blood vessels. As in other mammals, strongly GFAP positive astrocytes invade the ferret retina from the optic nerve. At birth, strongly GFAP positive astrocytes have reached about 22% of the distance between optic disc and outer retinal edge whereas weakly GFAP positive processes already extend to the edge of the retina. At postnatal days P30-P37 about 82% of the distance between optic disc and outer retinal edge and in the adult 88% of this distance is covered with strongly labelled astrocytes. Superficial blood vessels form from the optic disc. They reach up to about 24% of the retinal radius at birth and grow radially across the retina during further development. At P30-P37, the whole retina is covered with superficial blood vessels. The deep vascular layer forms later (around P30) through sprouting from superficial vessels. The radial pattern of astrocyte and vessel growth from the optic disc is not affected by the formation of the area centralis and visual streak. PMID- 11428048 TI - Hypertrophy of the ageing rat medial preoptic nucleus. AB - The medial preoptic nucleus (MPN) plays an essential role in the coordination of behaviours and physiological responses necessary for reproduction. Since ageing is associated with a progressive deterioration of reproductive functions we have explored the possibility that changes in the structural organization of the MPN might be implicated in this process. Thus, we have estimated the volume of the MPN, and the total number and size of its neurons, using stereological methods, and quantitatively evaluated the dendritic trees of MPN neurons in Golgi impregnated material. Male and female rats, aged 6, 24 and 30 months, were independently analysed. No cell loss was observed in aged rats of both sexes. However, the volume of the MPN and the somatic size of its neurons were remarkably enlarged in aged rats. No significant age-related changes in the size or shape of the dendritic trees or in dendritic spine density were found. To evaluate whether the changes observed in aged rats could be ascribed to an altered interaction between gonadal steroids and steroid-sensitive neurons, we have additionally estimated the total number of MPN neurons immunoreactive for the estrogen receptor-alpha. No significant age-related variations were detected. The age effects upon the MPN were more marked in females than in males and, consequently, the sexual dimorphisms in neuronal size and in the number of estrogen receptor-immunoreactive neurons were blunted in aged rats. PMID- 11428049 TI - Neuronal distribution and subcellular localization of HCNP-like immunoreactivity in rat small intestine. AB - A novel peptide, hippocampal cholinergic neurostimulating peptide (HCNP), originally purified from young rat hippocampus, affects the development of specific cholinergic neurons of the central nervous system in vitro. In this study, HCNP-like-immunoreactive nerve processes and nerve cell bodies were identified by electron microscopic immunocytochemistry in the rat small intestine. Labeled nerve processes were numerous in the circular muscle layer and around the submucosal blood vessels. In the submucosal and myenteric plexuses, some HCNP-like-immunopositive nerve cell bodies and nerve fibers were present. The reaction product was deposited on the membranes of various subcellular organelles, including the rough endoplasmic reticulum, Golgi saccules, ovoid electron-lucent synaptic vesicles in axon terminals associated with submucosal and myenteric plexuses, and the outer membranes of a few mitochondria. The synaptic vesicles of HCNP-like positive terminals were 60-85 nm in diameter. The present data provide immunocytochemical evidence that HCNP-like-positive nerve cell bodies and nerve fibers are present in the submucosal and myenteric plexuses of the rat small intestine. An immunohistochemical light microscopic study using mirror-image sections revealed that in both the submucosal and myenteric ganglia, almost all choline acetyltransferase (ChAT)-immunoreactive neurons were also immunoreactive for HCNP. These observations suggest (i) that HCNP proper and/or HCNP precursor protein is a membrane-associated protein with a widespread subcellular distribution, (ii) that HCNP precursor protein may be biosynthesized within neurons localized in the rat enteric nervous system, and (iii) that HCNP proper and/or HCNP precursor protein are probably stored in axon terminals. PMID- 11428050 TI - Expression of GFR alpha-1, GFR alpha-2, and c-Ret mRNAs in rat adrenal gland. AB - Glial cell line-derived neurotrophic factor (GDNF), an important factor for developing and lesioned pre- and postganglionic sympathetic neurons, and its congeners signal through a receptor complex consisting of the tyrosine kinase c Ret and a lipid-anchored alpha receptor (GFR alpha 1-4). Using in situ hybridization we show now that the mRNA for GFR alpha-2 is abundant in the adult rat adrenal medulla and its chromaffin cells. Coexpression of c-Ret and GFR alpha 1 mRNA's is restricted to a scarce subpopulation of medullary sympathetic neurons. Both GFR alpha-1 and GFR alpha-2 mRNA's are associated with preganglionic nerve trunks in the adrenal cortex. It is conceivable therefore that GDNF and related factors may activate chromaffin and preganglionic Schwann cells through a GFR-alpha receptor in absence of c-Ret. PMID- 11428051 TI - Differential effects of NgCAM and N-cadherin on the development of axons and dendrites by cultured hippocampal neurons. AB - A fundamental step in neuronal development is the acquisition of a polarized form, with distinct axons and dendrites. Although the ability to develop a polarized form appears to be largely an intrinsic property of neurons, it can be influenced by environmental cues. For example, in cell cultures substrate and diffusible factors can enhance and orient axonal development. In this study we examine the effects of growth on each of two cell adhesion molecules (CAMs), NgCAM and N-cadherin, on the development of polarity by cultured hippocampal neurons. We find that although the same pattern of development occurs on control substrates and the CAMs, the CAMs greatly accelerate the rate and extent of development of axons-axons from sooner and grow longer on the CAMs than on the control substrate. In contrast, the CAMs have opposite effects on dendritic development-N-cadherin enhances, but NgCAM reduces dendritic growth compared to control. These results provide further evidence that the development of polarity is largely determined by a cell-autonomous program, but that environmental cues can independently regulate axonal and dendritic growth. PMID- 11428052 TI - [Antiretroviral resistance testing. A necessary element in the follow-up of the HIV-infected patient]. PMID- 11428053 TI - [Comparative analysis of methods for detecting resistance]. PMID- 11428054 TI - [Primary resistance to antiretroviral agents]. PMID- 11428055 TI - [Resistance to nucleoside reverse transcriptase inhibitors]. PMID- 11428056 TI - [Biological basis for generating resistance to antiretroviral drugs]. PMID- 11428057 TI - [Resistance to non-nucleoside reverse transcriptase inhibitors]. PMID- 11428058 TI - [Resistance to protease inhibitors]. PMID- 11428059 TI - [Studies on resistance are useful in guiding antiretroviral therapy]. PMID- 11428060 TI - [HIV susceptibility determination: a qualified position against regarding a complex technology]. PMID- 11428061 TI - [Consensus documents and clinical guidelines on resistance to antiretroviral agents]. PMID- 11428062 TI - [GESIDA Consensus document on the use of resistance studies in clinical practice]. PMID- 11428063 TI - [Geriatrics--imbedding into the health care scheme of Germany]. AB - In this paper, we first give an overview of the origins of geriatrics. We then give a description of the development of modern geriatrics in the United States and Great Britain, based on the work of Nascher and Warren. Finally, we give an account of the development of geriatrics in Germany. Clinical geriatrics, which first developed 20-30 years ago, has shown a high rate of growth in the last few years. However, recently there has been a slowing down in this rate of growth--if not a complete halt or, indeed, a counter-movement. The reasons for this somewhat negative development in the past five years are comprehensively discussed. In the last part of the paper, improvements which could be made in geriatrics are described, thus possibly achieving a uniform and self-assured provision of geriatric care in the future, in relation to four priority areas, with regard to outpatient and inpatient care. The first priority area is the care of geriatric patients at home and in nursing homes. To this end, geriatrics in priority areas will be promoted. The second priority is the development of geriatrics in hospitals (acute geriatrics departments and departments with rehabilitation as their focuses of activity). A third priority is geriatric rehabilitation clinics. Finally, we need a few geriatric centers in Germany (centers of excellence), geriatric departments with professorial positions and university clinics with geriatric departments in which geriatrics is taught and from which the impulse for research will be given. PMID- 11428064 TI - [Epidemiological aspects of morbidity in the elderly]. AB - The spectrum of morbidity in older age is dominated by chronic diseases. Higher age is associated with functional difficulties, limitations, and disability. However, inter- and intraindividual variability is wide. Sequela of chronic diseases are crucial to the development of dependency and the use of health care. They are cost intensive. The number of very old people will particularly increase according to estimations of future demographic changes in Germany. It can, thus, be assumed that the need for geriatric care, specific prevention and rehabilitation will not decrease soon. PMID- 11428065 TI - [Overview of health care services in clinical and rehabilitation geriatrics and their development in Germany]. AB - A survey of the 2000 statistics about the existing geriatric care structures in Germany showed that the number of inpatient geriatric care centers rose from 84 in 1993 to 318 in 2000. The number of day clinics rose in the same timeframe from 13 to 136. However, there are serious deficits in the area of ambulant geriatrics; in 2000 there were only 11 such institutions. The question is discussed as to why there are three different care structures for acute geriatrics, neurological early rehabilitation, and general early rehabilitation in hospitals. It is suggested how the existing care structures could be sensibly integrated. PMID- 11428066 TI - [Ambulatory geriatric rehabilitation--assessment of current status and prospects]. AB - Clinical geriatrics in Germany has evolved remarkably since 1990, when first concepts had been developed for establishing structures and facilities for the ambulatory rehabilitation of geriatric patients. However, achievements during the last 10 years were made only for inpatient geriatrics and geriatric day clinics. Geriatric outpatient rehabilitation (GOR) is still a "missing link" in the networked care for elderly patients in Germany and remains until today in a planning phase. The article is aimed at describing the current organisational status of geriatric rehabilitation in Germany and focuses on the question of where and how to fit in geriatric outpatient rehabilitation. Distinction criteria between the different types of geriatric rehabilitation as well as indications, goals, and organisational aspects of GOR are discussed. PMID- 11428067 TI - [Is the ICIDH (International Classification of Impairments, Disabilities and Handicaps) suitible for geriatric rehabilitation?]. AB - The International Classification of Impairments, Disabilities, and Handicaps (ICIDH), a manual of disease consequences issued by WHO, at present exists in two versions. ICIDH-1, issued in 1980, has been under revision since 1993. This revision process will result in a more sophisticated and more comprehensive ICIDH 2 in 2001. The ICIDH, based on the biopsychosocial disease model, describes disease consequences in three dimensions: Structural and functional body impairments. Disabilities (ICIDH-1) or restrictions of activities (ICIDH-2) in activities of daily living. Handicaps (ICIDH-1) or participation limitations (ICIDH-2) in social integration. In the field of geriatrics ICIDH is also helpful as a coherent conceptual framework and reference manual to determine and code the disabilities and handicaps with regard to daily living competence. Geriatric assessment instruments are no alternative, but complementary to the ICIDH. In view of the high frequency of geriatric patients with multimorbidity, i.e. multiple impairments, disabilities, and handicaps, the utility of the ICIDH in geriatric care is highly dependent on the flexible application of this classification system. PMID- 11428068 TI - [Geriatric assessment. The status of current knowledge with reference to suitability criteria (discrimination, prediction, evaluation, practical aspects)]. AB - Comprehensive geriatric assessment was developed in the Anglo-American countries as a diagnostic process to better understand the effects of diseases and interactions of multiple chronic conditions. To standardize the use of assessment instruments, a working group of the two German geriatric societies has elaborated recommendations. As the first step, an expanded version of the screening according to Lachs should be used, followed by performing the Barthel-ADL, the Mini-Mental State Examination, the Geriatric Depression Scale, the 'Timed Up and Go' and the (Semi-) Tandem Stance. The expanded version of the screening of Lachs is helpful for targeting patients and for placement; the others are not useful for that purpose. No instrument satisfactorily fulfills all test criteria, which include validity, reliability, sensitivity, practicability and repeatability, and, in part, these dimensions are not investigated at all. All test instruments are diagnostic tools for functional status and capabilities but do not give information about resource needs. No instrument covers the new concept of ICIDH completely. PMID- 11428069 TI - [Education, graduate and continuing education in geriatrics and geriatric rehabilitation]. AB - Education and training in geriatric medicine for family doctors and medical specialists, nurses, physio- and ergotherapists, social workers becomes more important. A curriculum in high quality geriatric medicine can help solve the problems coming up with the increasing life-expectancy and with the increasing proportion of elderly depending on care and social services. The development of curricula and programs in high quality geriatric medicine demonstrates that the geriatrician has a special approach to the elderly patient different from traditional medicine. Describing and teaching this special approach to the elderly patient shows the special view of the geriatrician working with elderly patients. A number of barriers to program development must be considered. To successfully implement geriatrics training, geriatric faculties need to have adequate protected time to plan, administer and teach the curriculum. It is very important to implement geriatrics training in universities and medical institutes. PMID- 11428070 TI - [Expert assessment of nursing care need according to SGB XL. Trigger for intervention programs in geriatric rehabilitation]. AB - "Rehabilitation instead of nursing care" is one of the most important entitlements of the social welfare legislation to long-term care. In Germany 1.2 million evaluations per year are done by the experts of the medical review boards of the health insurance funds. As a part of those evaluations the experts also have to discuss the necessity of any kind of rehabilitation procedures. In 1999 according to the experts' opinion only about 12% of the patients need such special support. At first this number seems much too low. We discuss the problems in that social field: Does the demand of the social welfare legislation "prevention and rehabilitation instead of nursing care" not correspond to the real needs or are the circumstances of social policy and medical supply responsible for that low recommendation of rehabilitation programs. This discussion leads to the following consequences: The patients, the members of their family but also the treating physicians must be informed much better than today about diagnostic and therapeutic possibilities for the special diseases of elderly people. All in the game have to prepare concepts, pilot projects and then common solutions for geriatric rehabilitation programs as soon as possible. Politicians, physicians, nursing staff, rehabilitation staff, patients, family members and any other caregivers all have to take responsibility, maybe the liability. Not only German people will be growing older than ever before, which everyone has to recognize as one of the most important facts of our welfare state. To handle this development will be the most serious duty of our community in the future. Therefore, geriatrics has to receive high public attention. PMID- 11428071 TI - [Health services research in geriatrics and geriatric rehabilitation from the national and international viewpoint]. AB - In Germany, health services research geriatric problems is not of major scientific and political importance. Focusing on geriatrics, it is essential to produce good data concerning efficacy, effectiveness and costs of the interventions. Otherwise, the discussion about the allocation of resources in the German health care system will take place without good arguments for special geriatric interventions. Prerequisite for this goal is the definition of adequate endpoints targeting functional deficits and the operationalization of multi- and comorbidity. Another problem is research focused on the "black box" of specific geriatric interventions. A special German situation is the distinction between acute and rehabilitative geriatric settings where very similar patients are treated. For this reason, it is essential for the geriatrician in the German context to focus more on health services research to demonstrate with scientific evidence their important contribution for the care of the elderly. PMID- 11428072 TI - [Referral and management in geriatrics and geriatric rehabilitation with special reference to medical case fee schedules and diagnosis related groups (DRGs)]. AB - Every system of payment of flat-rate case charges results in an economically determined optimizing of treatment processes. This presupposes a high level of co operation between all those involved in the treatment process. Collaboration must extend over occupational groups (doctors, nursing staff and therapists), over departments (specialist geriatric competence and specialist knowledge) and must increasingly take place in a manner extending over sectors (inpatient, partly inpatient and outpatient). To this end, interdisciplinary, team-oriented modes of working provide the direction to be followed in geriatrics. What is essential for efficient patient management is for patient allocation control to be implemented in good time, with individual requirements regarding treatment being provided by the most suitable department. This will only be possible, in perspective, by implementing binding treatment guidelines and quality standards, as well as the timely interdisciplinary incorporation of acute geriatrics, e.g. by participation of the admission wards of the clinics. Geriatrics in acute hospitals has demonstrated its value and has become an indispensable element of appropriate health provision in Germany. The retention of this structure of geriatric care presupposes that, in the future, the DRG system of geriatric services will also be included with a sufficient measure of differentiation and that consequently an appropriate level of remuneration for geriatric services will be ensured. As the DRG system has not been applied internationally, up until now, in the area of acute geriatrics, suitable adjustments to the system of the basic Australian AR DRG system are absolutely essential. PMID- 11428073 TI - [Development of geriatrics in Germany]. AB - After an inquiry of the Bundesministerium fur Arbeit und Sozialordnung by the responsible Landesministers, the number of inpatient geriatric institutions rose by about 34% from 1997 to 2000 and the number of partly inpatient geriatric institutions rose by 106% in the same timeframe. Outpatient geriatric rehabilitation institutions are, on the other hand, still basically nonexistent, whose number has risen from 9 to 12 since 1997. The situation is similar for mobile outpatient services, whose number doubled from 5 to 10. PMID- 11428074 TI - [Glucocorticoid-induced osteoporosis. A underestimated sequela of long-term treatment of autoimmune diseases]. AB - The treatment of autoimmune diseases with systemic glucocorticosteroids remains a therapeutic challenge and requires close collaboration with internists, radiologists and in some cases orthopedic surgeons. Generally, patients initially receive high-dose glucocorticosteroid therapy and are then treated for a longer period of time with moderate to low doses above the level causing symptoms of Cushing's disease. A major cause of complications is glucocorticosteroid-induced osteoporosis with hip fractures, crush fractures of the spine and other low trauma fractures as well as deformities of the skeleton leading to neurological and other systemic problems. Loss of bone mass as a result of high dose or long term systemic glucocorticosteroid treatment is well studied and can be documented in a standardized and reproducible fashion using modern radiological techniques. In recent years several controlled studies of bone loss and therapy of osteoporosis have been published, mostly including patients with rheumatoid arthritis or systemic lupus erythematosus. This review discusses recent publications and provides a brief overview on therapeutic options. PMID- 11428075 TI - [Guideline for evaluating drugs in chronic venous insufficiency. 10 May 2000 edition]. PMID- 11428076 TI - [Treatment of therapy refractory verrucae vulgares with a ciclopirox-containing lacquer]. AB - BACKGROUND AND OBJECTIVE: Anti-inflammatory actions of ciclopirox, an antifungal agent, have been described previously. We assessed the effectiveness of ciclopirox in the treatment of viral warts. PATIENTS/METHODS: Twenty-three immunocompetent patients (age 9-61 years) with common warts at the hands, feet and in the face resistant to conventional therapy were treated. Following keratolysis with a salicylic acid-containing patch, an 8% ciclopirox lacquer was applied 1-2 times daily. At the beginning and the end of the treatment lesions were photographed and the time span (minimum 4 weeks) until complete resolution was documented. RESULTS: In 7 patients complete remission, and in 12 patients partial remission was achieved. Only 4 patients did not show any effect under topical ciclopirox treatment (mean duration of therapy 3.2 +/- 2.1 months). The overall response rate was 82.6%. Local side effects like erythema or pruritus were not reported. CONCLUSION: Our data suggest that a ciclopirox lacquer may be helpful in the treatment of common warts. However, the data have to be confirmed in a randomized controlled trial in the future. PMID- 11428077 TI - [Development of the German Scale for Assessing Quality of Life in Skin Diseases]. AB - BACKGROUND AND OBJECTIVE: A German dermatological quality-of-life (QoL) instrument was developed on the basis of a representative and standardised patient population. PATIENTS/METHODS: Qualitative interviews were performed in 633 patients. Based on the answers of 20 patients from each of the 10 most frequent diagnostic groups, items were identified and a questionnaire developed. This was used in 704 patients and eventually the number of items was reduced. RESULTS: The long version of the instrument has 72 items. The median of the total score of the questionnaire was 75 and significantly different between diagnostic groups (herpes/shingles 110, verruca 26; p < 0.001). Significant differences occurred also between questionnaire domains with categories "psychological, physical, and treatment" showing the highest results. In contrast to the total group, patients with urticaria scored highest in dimensions "social, leisure, and work" (p < 0.001). By factor analysis the number of items was reduced to 36. CONCLUSIONS: The DIELH was developed on the basis of a standardised and representative patient population. The long version proved to have a good discriminant validity with respect to diagnostic groups and dimensions. PMID- 11428078 TI - [Successful symptomatic tazarotene treatment of juvenile acanthosis nigricans of the familial obesity-associated type in insulin resistance]. AB - An 11-year-old boy suffering from morbid obesity since infancy developed at age 9 a progressive brown-black hyperpigmentated and hyperkeratotic eruption in the neck and axillary region with minor involvement of both groins. Based on this clinical picture, and confirmed by histopathology, we diagnosed acanthosis nigricans. Following a thorough endocrinological examination and because the patient's obese mother showed similar skin lesions, the disease was subclassified as a familial obesity-associated type of acanthosis nigricans associated with insulin resistance. In a right-left-comparison the affected skin of one body side was treated with tazarotene 0.05% versus urea 10%, once daily each. A great benefit for the tazarotene-treated over the opposite side could already be seen after three weeks which was also verified by dermatohistopathology. Three months after topical tazarotene treatment had been extended to both sides, the residual lesions were significantly improved. The highly satisfying, good result has been maintained up to now by a continuous topical retinoid treatment over 18 months, usually with an interval application regimen, i.e., 3 x per week. PMID- 11428079 TI - [Pemphigus vegetans. A historical perspective]. AB - Pemphigus vegetans is a rare variant of pemphigus which is mainly localized in the intertriginous areas (Neumann type, Hallopeau type) or primarily involves the mucous membranes (pyostomatitis vegetans). A 18-year-old patient with erosive stomatitis developed a vegetating plaque with papillomatous and verruciform features in her left axilla. Histopathological examination of the axilla revealed papillomatosis and acanthosis as well as suprabasal clefting with acantholytic cells. By direct immunofluorescent examination, deposits of immunoglobulin IgG and complement (C3) were found in the intercellular space of the epidermis. Serological examination by indirect immunofluorescent techniques was indicative of pemphigus autoantibodies at a titer of 1:40. This case report demonstrates that the classical differentiation of pemphigus vegetans in two types, Neumann type and Hallopeau type, is mainly of historical importance, because both entities may represent variants of the same disease. In addition, other disorders such as IgA pemphigus may also present with vegetating plaques in intertriginous sites. PMID- 11428080 TI - [Skin manifestations of Erdheim-Chester disease. Case report and review of the literature]. AB - A 46 year old woman suffering from Erdheim-Chester disease is reported for the first time in the German dermatologic literature. She showed periorbital xanthelasmas and indurated erythemas on both flanks. Histologically the skin manifestations were also a sign of the basic disease which had spread to various organs. Small granulomas with foam cells, macrophages and few giant cells were found in the corium. Immunohistochemistry demonstrated positive reactions to protein S-100 and alpha-1-Antichymotrypsin, while CD 1 a-antigen and PNA were negative. Further localizations of Erdheim-Chester disease were found in the femurs, tibiae and mandibula as well as in the right breast, retroorbital region and abdominal aorta. Infiltration of the retroperitoneal cavity led to urinary retention and nephrectomy. With systemic corticosteroid therapy, the skin lesions on the flanks regressed but recurred after discontinuance of the drug. Erdheim Chester disease is now classified as part of the histiocytoses which is confirmed by our immunohistochemical findings. The histiocytes are probably not identical with Langerhans cells. The diagnosis is usually made by the radiologist on the basis of typical osteosclerotic symmetrical lesions of the long bones, histologically representing lipoid granulomas. Common non-osseous manifestations of the disease are in the retroperitoneal region, orbita and intracranially (partially associated with diabetes insipidus) which are often fatal. On the skin, xanthelasma and xanthoma are present in 1/6 of the cases; in some patients yellow-brown papular and widespread infiltrated lesions have been described. As regards to therapy corticosteroids chemotherapy, radiation and immunomodulating agents as well as surgery have been used with limited success. PMID- 11428081 TI - [Subcutaneous calcifications after radiotherapy]. AB - A 75-year-old female patient presented with late stage cutaneous radiation syndrome, following postoperative combined radiotherapy for cervical carcinoma administered 32 years ago. She was hospitalized because of a deep abscess in the radiation-exposed area on the sacrum. Extension into the subcutaneous fibrosis was verified by 7,5-MHz-sonography and nuclear magnetic resonance imaging. With intravenous cefotiam (Spizef) 2.0 g three times daily, the inflammation decreased, as seen clinically and with nuclear magnetic resonance imaging. As a secondary finding, an unusual distinctive subcutaneous calcification was diagnosed in the radiation-exposed area by means of 7,5-MHz-sonography, as well as computer- and nuclear magnetic resonance imaging. These subcutaneous calcifications are most likely to have been radiation-induced. PMID- 11428082 TI - [Multiple miliary osteomata cutis. Excision with "front lift" approach]. AB - Multiple miliary osteomata of the skin commonly occur on the face and the upper trunk of young healthy women. The mechanism of bone formation is unknown. There is no adequate therapy, which reduces or reverses the growth of osteomatas. Those tumours which are cosmetically disturbing may be excised by a small incision through the skin. For the first time we report their removal via the subcutaneous "front-lift" approach. PMID- 11428083 TI - [Large nodular lymphomatoid papulosis associated with parakeratosis variegata]. AB - A 63 year old man suffering from lymphomatoid papulosis in association with parakeratosis variegata over a period of 20 years is presented. This case is unusual in respect to long-standing history, the rare combination of the two entities, and the extraordinary size of the nodules of lymphomatoid papulosis. Extracorporal photopheresis has induced partial remission lasting up to nine months so far. PMID- 11428084 TI - [Familial dyskeratotic comedones. A rare entity]. AB - Familial dyskeratotic comedones is a rare autosomal dominant genodermatosis. In childhood or adolescence disseminated keratotic papules develop and gradually increase in number with time. The isolated papules show a central keratotic plug which tends to recur after extraction. Pruritus and occasional inflammation are the only symptoms. The lesions appear on the extremities, and less frequently on the trunk and the face. About half of the patients have a history of acne vulgaris. The histologic picture is highly characteristic and shows a deep invagination of an acantholytic and dyskeratotic epidermis with prominent cornification. Familial dyskeratotic comedones are generally refractory to any therapy. We report on two sisters with familial dyskeratotic comedones successfully treated by CO2-laser therapy. PMID- 11428085 TI - [Human experimentation in dermatology 100 years ago and today]. AB - The problem of ethics in dermatology is explored using selected examples from the literature. The classic case of Neisser as well as Eduard Arning's experiments on a Hawaiian with leprosy convicted to death are discussed. In addition, inoculation studies with syphilis 100 years ago and the author's own experiences are cited. The differences in the approaches to medical ethics 100 years ago and today lead to the conclusion that ethics change to fit the needs of the society and do not have a permanent fixed status. PMID- 11428086 TI - [Comment on the contribution by S. Stander, K. Assmann, D. Nashan, B. Wigbels, T. Luger, D. Metze: "Uninterrupted incision margin controlled surgery with paraffin section of melanomas of the face"]. PMID- 11428087 TI - [Comment on the contribution by S. Stander, K. Assmann, D. Nashan, B. Wigbels, T. Luger, D. Metze: "Uninterrupted incision margin controlled surgery with paraffin section of melanomas of the face"]. PMID- 11428088 TI - [Pediatric dermatology in Mexico--a stipend sojourn]. PMID- 11428089 TI - [Figurated plaque on actinically damaged skin. Actinic O'Brien granuloma]. PMID- 11428090 TI - [Therapy of varicella zoster and herpes simplex virus-induced diseases. 2: References for implementing and indications for virustatic therapy]. PMID- 11428091 TI - [Information Society of Dermatological Clinics. Clinical epidemiology for prevention of allergic contact eczema]. PMID- 11428092 TI - ["One does not snack on prions!" About illnesses, cannibalism and cosmetics]. PMID- 11428093 TI - [Diagnosis related groups (DRG). Perspectives, analysis, strategies from the viewpoint of dermatology]. PMID- 11428094 TI - [Phenotypic variations of the genotype: retrotransposons as epigenetic mediators?]. PMID- 11428095 TI - [Emergence of drug-resistant gram-positive pathogenic bacteria]. AB - Staphylococcus aureus is a common cause of soft tissue infection, e.g. impetigo, cellulitis, or wound infection, and causes osteomyelitis, arthritis, bacteremia with metastatic infection, and scalded skin and toxic shock syndromes. Coagulase negative staphylococci have become increasingly important causes of nosocomial bacteremia associated with invasive monitoring, intravascular catheters and prosthetic heart valves or joints. Most staphylococci produce blactamase and are resistant to penicillin. An increasing proportion of S. aureus have intrinsic resistance to methicillin (MRSA) and present major problems in hospitals for the control of cross infection. The glycopeptides, teicoplanin and vancomycin, are the antibiotics of first choice for treatment of these infections. After the first report describing a Japanese clinical isolate of vancomycin-resistant S. aureus (VRSA), several papers have documented the emergence of these microorganisms. Since the development and spreading of this phenomenon which is perceived as a fearsome threat to the already difficult therapy of nosocomial infections due to the prevalence of heterogeneous vancomycin resistance, we found the incidence of MRSA exceeds 35% in our hospital. Out of 179 methicillin resistant S. aureus isolated during 1997-1998, two strains (1.1%) gave subclones with vancomycin MICs of 8 mg/L. PFGE showed identical restriction patterns for both isolates, suggesting transfer of a single clone between two different patients. PMID- 11428096 TI - [Risk factors and prevention of infections of hip and knee prostheses]. AB - Prevention of deep periprosthetic infection requires appreciation of the contributing factors to this phenomenon. The concept of an interdependent relationship between the bacteria, wound and host is most useful when considering the prevention of infections. Establishment of infection depends on the number and virulence of the bacteria, the host's ability to eliminate those bacteria and the status of the wound. Multiple variables contribute to the deposition and proliferation of bacteria into the wound environment which enhance the infectious process, such as the presence of a prosthetic implant or medical conditions and medications which impair the patient's defense. Prevention must address optimization of the wound environment, minimizing the number of bacteria into the wound and augmenting the host defenses. Administration of prophylactic antimicrobials prior to skin incision reduces the incidence of wound infections. The optimal antimicrobial agent should have excellent in vitro activity against staphylococci and streptococci, a long serum half-life, good tissue penetration, be non-toxic and inexpensive. PMID- 11428097 TI - [Antibiotic chemoprophylaxis in orthopedic prosthesis implantation]. AB - In elective orthopedic surgery, as well as in any type of clean operations, it is now advised to administer antibiotic prophylaxis only when extraneous material is implanted (prostheses). Given the most frequent appearance of staphylococcal postoperative infections, in orthopedic surgery the antibiotic used for prophylaxis should be particularly active against these microorganisms. Therefore, the first- (cefazolin) and second-generation (cefuroxime or cefamandole) cephalosporins are usually recommended. Because prosthetic infections are more and more frequently caused by methicillin-resistant staphylococci, the glycopeptides (vancomycin and teicoplanin) are often recommended for prophylaxis and more frequently used in routine practice, at least in hospital environments in which these microorganisms often appear. Indeed, various investigations have demonstrated that a single dose of teicoplanin is equally protective as multiple doses of other antibiotics usually used. Regardless of the type of surgery, perioperative prophylaxis should be administered as an i.v. bolus or as a small infusion. However, for mono- or bilateral knee prosthesis, regional prophylaxis has been suggested in recent years, i.e., antibiotic administration in a vein of the foot or leg being operated. The pharmacokinetic and clinical results of this technique have proved to be quite successful. PMID- 11428098 TI - [Antibiotic prophylaxis of postoperative infection in orthopedics. Results of an epidemiologic survey in Italy conducted by the Journal of Chemotherapy]. AB - An epidemiological survey of the use of antimicrobial prophylaxis in Italian hospitals was carried out under the auspices of the Journal of Chemotherapy. Out of 500 Italian orthopedic centers requested, 225 have participated in this study. A total of 136,321 surgical procedures were reported in the 166 centers reporting complete answers on type of surgery. They comprised hip and knee prosthesis (13.9%), spine surgery (4%), hip endoprosthesis (5.2%), osteosynthesis (26.9%), arthroscopy (24.4%), and others (25.5%). Perioperative antimicrobial prophylaxis was used in 75% of operations (ranging from 57.1% to 99.4% in arthroscopy and joint prosthesis, respectively). Short term (< 24 h) antimicrobial prophylaxis was performed in 38.4% of the 206 Centers answering this question correctly. 61.1% of Centers employed single agent prophylaxis. 70.8% of these prescriptions were betalactam antibiotics. Bacteriological analysis of the wound in 86 Centers (total number of isolates 2013) revealed the presence of Gram-positive isolates in 73.7% of cases. Methicillin resistance was expressed in 45% of 915 tested strains. Out of 4221 patients with high risk of infectious complications (joint prosthesis surgery) given antimicrobial prophylaxis in 46 Centers, the percentage of surgical wound infections was overall 2.1%, while that of non-surgical wound infections was 3.6%. A decrease in the total infection rate by about half was associated with long-term (> 24 h) as compared to short-term (< 24 h) antibiotic treatment (3.7 vs 7.6%, respectively), and with the use of antibiotic drug combinations vs single antibiotic drugs (3.9 vs 6.6%, respectively). The incidence of surgical-site infection is not decreased by extending the chemoprophylaxis for more than the first 24 h after surgery, while it is reduced from 2.5 to 1.4% by use of combination antibiotic therapy. PMID- 11428099 TI - [Introduction: current possibilities for the control of surgical infections in orthopedics]. PMID- 11428100 TI - Direct hydrolytic route to molecular oxo-hydroxo lanthanide clusters. PMID- 11428101 TI - A redox reaction to synthesize nanocrystalline Cu2-xSe in aqueous solution. PMID- 11428102 TI - Preparation, characterization, X-ray crystal structure, and energetics of cesium 5-cyano-1,2,3,4-tetrazolate: Cs[NCCNNNN]. AB - Cesium 5-cyano-1,2,3,4-tetrazolate, Cs[NCCNNNN] (Cs1), was prepared in 100% yield by a 3 + 2 cycloaddition reaction of CsN3 and (CN)2 in SO2. Cs1 forms monoclinic crystals (a = 8.297(2) A, b = 11.040(3) A, c = 6.983(2) A, beta = 120.31(2) degrees, space group C2/c, Z = 4, R1 = 0.048, wR2 = 0.120 for 1217 independent reflections). Cs1 is best described as a three-dimensional array of cations and anions connected by weak Cs(+)-N delta- contacts. The cations and anions each form a diamond-type lattice (tetrahedral arrangement of ions) with the counterions lying in hexagonal channels running parallel to the c-axis. The anions in the channels form stacks with the CN groups pointing in opposite directions in adjacent layers. The calculated (RB3PW91/6-311 + G*) geometry of 1 is in agreement with the X-ray crystal structure, and the calculated vibrational spectrum is in good agreement with the observed FT-IR and FT-Raman spectra. The lattice enthalpies and heats of formation of M1 (M = Cs, K) as well as the enthalpy of formation from MN3(s) and (CN)2(g) were estimated. The 13C and 14N NMR spectra of Cs1 are also reported. The ionization potential (450.7 kJ mol) and electron affinity (427.4 kJ/mol) of 1 were calculated. Attempts to oxidize 1 with AsF5 led to the formation of Cs1.xAsF5 (x approximately 2). The 7 pi dianion [NCCNNNN].2- is calculated to be a stable entity in the gas phase, but Cs(2)1 is estimated to be unstable with respect to dissociation to 2 CsCN and 3/2 N2 (delta Hdiss = -132.4 kJ/mol). The preparation of the potassium salt of 1 and the corresponding thermodynamic quantities are reported. PMID- 11428103 TI - Structure of triamidoaluminum complexes: a theoretical ab initio/IMOMM study. AB - The electronic structure of three-coordinated complexes of aluminum Al(NR2)3 has been studied through theoretical calculations. In the unsubstituted system Al(NH2)3, ab initio calculations (MP2/6-31G(d,p) level) show that the optimal geometry results from a conrotatory motion of the three amido substituents starting from the fully conjugated planar species. The energy difference between these two structures is found to be small (less than 0.5 kcal/mol). In methylated species Al(NMe2)3, steric effects become important and the planar geometry is destabilized. The conrotatory geometry corresponds to the absolute minimum and is located 14.2 kcal/mol below the planar structure (MP2/6-31G(d,p) level). Several coupled motions of the amido substituents have been computed and have been found to stabilize the system with respect to the fully conjugated structure. A rough estimate of the steric repulsion is calculated by comparison between the unsubstituted and methylated species. Finally, the real molecules Al[N(SiMe3)2]3 and Al(NiPr2)3 have been studied through a coupled quantum mechanics/molecular mechanics method. In accordance with the experimental data, it is found that the conrotatory minimum is the absolute minimum in the R = SiMe3 case whereas a less symmetrical minimum is found in the R = iPr case. In this last minimum, an amido group is almost deconjugated and the two other groups move in a conrotatory manner. The different behavior of these two systems may originate from the quasi spherical shape of the SiMe3 group, which leads to unavoidable steric repulsion. PMID- 11428104 TI - Alkaline earth complexes of 1,4,7,10-tetrakis(2-hydroxyethyl)- and 1,4,7,10 tetrakis(2-methoxyethyl)-1,4,7,10-tetraazacyclododecane: an equilibrium and kinetic study. AB - The 1,4,7,10-tetrakis(2-hydroxyethyl)-1,4,7,10-tetraazacyclododecane complexes [M(thec12)]2+, where M2+ = Mg2+, Ca2+, Sr2+, and Ba2+, are characterized by log(K/dm3 mol-1) = 2.86 +/- 0.09, 7.41 +/- 0.04, 6.47 +/- 0.04, and 4.84 +/- 0.03 at 298.2 K in aqueous Et4NClO4 (I = 0.10 mol dm-3), where K is a potentiometrically determined stability constant. The analogous literature values for the 1,4,7,10-tetrakis(2-methoxyethyl)-1,4,7,10-tetraazacyclododecane complexes [M(tmec12)]2+ are 2.47, 5.47, 5.00, and 4.72. The enantiomerization of eight-coordinate delta- and lambda-[M(thec12)]2+ is characterized by k(298.2 K) = 2310 +/- 260, 582 +/- 17, and 445 +/- 5 s-1, delta H++ = 19.1 +/- 0.8, 33.3 +/- 0.5, and 43.9 +/- 0.4 kJ mol-1, and delta S++ = -117 +/- 4, -80.3 +/- 1.8, and 47.0 +/- 1.3 J K-1 mol-1 when M2+ = Mg2+, Ca2+, and Ba2+, respectively, in methanol-12C-d4 as shown by 13C NMR spectroscopy. For the enantiomerization of eight-coordinate delta- and lambda-[M(tmec12)]2+, k(298.2 K) = 310 +/- 1 and 688 +/- 3 s-1, delta H++ = 54.0 +/- 0.2 and 39.6 +/- 0.1 kJ mol-1, and delta S++ = 16.1 +/- 0.5 and -57.9 +/- 0.3 J K-1 mol-1 when M2+ = Ca2+ and Ba2+, respectively. However, [Mg(tmec12)]2+ has a seven-coordinate structure where one of the methoxy groups is not coordinated and exchange of the methoxy groups between the coordinated and free states is characterized by k(298.2 K) = 163,000 +/- 8000 s-1, delta H++ = 35.8 +/- 0.4 kJ mol-1, and delta S++ = -25.1 +/- 1.7 J K-1 mol-1. The intermolecular exchange of thec12 and tmec12 between the coordinated and free states is substantially slower than the enantiomerizations in the first five complexes and the intramolecular exchange process observed in [Mg(tmec12)]2+. PMID- 11428105 TI - Wide spectral range nonlinear optical crystals of one-dimensional coordination solids [Et4N][Cd(SCN)3] and [Et4N][Cd(SeCN)3] and the general design criteria for [R4N][Cd(XCN)3] (Where R = Alkyl and X = S, Se, Te) as NLO crystals. AB - We report herein two new nonlinear optical (NLO) crystals, [Et4N][Cd(XCN)3], where X = S (1) and Se (2), that are transparent from 220 to 3300 nm, covering the entire near-ultraviolet, the visible, and the near-infrared spectral regions and giving rise to a very wide and continuous optical window, which is useful for many frequency conversion applications. Both 1 and 2 exhibit highly efficient second harmonic generation (SHG) as measured via the Kurtz-Perry method. The corresponding [Me4N]+ salts, [Me4N][Cd(XCN)3 where X = S (3) and Se (4), show no SHG effects. All four structures adopt noncentrosymmetric space groups (Cmc2(1) for 1 and 2 and Pna2(1) for 3 and presumably 4) and are based on one-dimensional anionic [Cd(XCN)3-] infinity zigzag chains. However, a detailed analysis of the structures of [R4N][Cd(XCN)3], where R = Et, Me and X = S, Se revealed that the difference in the second-order nonlinear responses of the Et4N+ salts (1 and 2) and the Me4N+ salts (3 and 4) is attributable to the relative alignment of the [Cd(XCN)3-] infinity zigzag chains, being parallel in the crystals of 1 and 2 but antiparallel in the crystals of 3 and 4. Also reported, for the first time, are the synthesis and crystal structure of [Et4N][Cd(SeCN)3] (2). Compound 2 crystallizes in an orthorhombic unit cell of Cmc21 space group symmetry with lattice parameters of 9.938(1) A, 16.868(2) A, 11.054(1) A, and Z = 4. Other issues related to the molecular and crystal engineering of this class of NLO materials are also discussed. PMID- 11428106 TI - Synthesis, structure, and magnetic characterization of a hysteretic charge transfer salt metamagnet, decamethylferrocenium 2,3-dicyano-1,4-naphthoquinonide, [Fe(Cp*)2][DCNQ]. AB - The synthesis and single-crystal structure of a new one-to-one charge-transfer salt, derived from decamethylferrocene and 2,3-dicyano-1,4-naphthoquinone, are described. [Fe(Cp*)2][DCNQ] crystallizes in the orthorhombic space group Pbca, with a = 17.3149(5) A, b = 14.6862(4) A, c = 21.0507(6) A, and Z = 8. Magnetization vs temperature data obtained in 100 G suggest that the compound exhibits dominant one-dimensional ferromagnetic coupling and that it subsequently undergoes an antiferromagnetic phase transition below TN approximately 4 K. Results of magnetization vs applied field experiments show that the compound is a metamagnet with a critical field of approximately 3 kG at 1.8 K. In the nominally antiferromagnetic state, apparent canting of the moments gives rise to a small amount of hysteresis. This picture is supported by ac susceptibility data. The 57Fe Mossbauer spectrum exhibits the expected decamethylferrocenium unresolved quadrupole doublet (delta = 0.53 mm/s) at 77 K and magnetic hyperfine splitting, Hint = 37.9 T, corresponding to long-range magnetic order at 1.63 K. PMID- 11428107 TI - Solution equilibria of tetrakis(ethyisocyanide)platinum(II) with tetracyanoplatinate(II): equilibria and thermodynamics of the formation of di-, tri-, and tetraplatinum species. AB - In aqueous solution, the tetrakis(ethylisocyanide)platinum(II) cation, Pt(CNC2H5)4(2+), reacts with tetracyanoplatinate(II), Pt(CN)4(2-), to form four multinuclear species. These species are the diplatinum(II) compound [Pt(CNC2H5)4(2+)][Pt(CN)4(2-)], two triplatinum compounds, [Pt(CNC2H5)4(2+)]2[Pt(CN)4(2-)]2+ and [Pt(CNC2H5)4(2+)][Pt(CN)4(2-)]2(2-), and the tetraplatinum species [Pt(CNC2H5)4(2+)]2[Pt(CN)4(2-)]2. The interactions leading to the stability of these species are not simply ionic. The interactions result in major differences in the electronic absorption spectra, and the oligomers are not disrupted by the addition of other salts. The formation constant for each of the oligomers was determined at several temperatures, and the enthalpies and entropies of formation were measured. Species with even numbers of platinum ions have more negative stepwise enthalpies of formation than do the species with odd numbers of platinum ions. Entropies of formation are negative as well; however, the values are significantly more positive for the species with odd numbers of platinum ions than for those with an even number. These thermodynamic trends can be rationalized by recognizing that the species with even numbers of platinum ions are dipolar and organize the solvent better than the species with odd numbers of platinum ions, which are not dipolar. Thus, the more negative enthalpies of formation of the species with even numbers of platinum ions are a consequence of better solvation than species with odd numbers of platinum ions. Accordingly, the entropies of formation of the species with even numbers of platinum ions are more negative because they organize solvent to a greater extent. Exclusive of solvent effects, the anion-cation interaction is about 30 kJ/mol. PMID- 11428109 TI - Synthesis, crystal structure, EXAFS, and magnetic properties of catena [mu tris(1,2-bis(tetrazol-1-yl)propane-N1,N1')iron(II)] bis(perchlorate). First crystal structure of an iron(II) spin-crossover chain compound. AB - [Fe(btzp)3](ClO4)2 (btzp = 1,2-bis(tetrazol-1-yl)propane) represents the first structurally characterized Fe(II) linear chain compound exhibiting thermal spin crossover. It shows a very gradual spin transition (T1/2 = 130 K) which has been followed by magnetic susceptibility measurements and 57Fe Mossbauer spectroscopy. The structure has been solved at 200 and 100 K by single-crystal X-ray analysis. It crystallizes in the trigonal space group P3c1 with Z = 2 Fe(II) units at both temperatures. The molecular structure consists of chains running along the c axis in which the Fe(II) ions are linked by three N4,N4' coordinating bis(tetrazole) ligands. The main difference between the two forms appears to be in the Fe-N bond lengths, which are 2.164(4) A at 200 K and 2.038(4) A at 100 K. The Fe-Fe separations are 7.422(1) A at 200 K and 7.273(1) A at 100 K. The EXAFS results are consistent with the crystal structure. In both spin states, the FeN6 octahedron is almost regular within the EXAFS resolution. The Fe-N distance is found as 2.16(2) A at 300 K and 2.00(2) A at 40 K. The absence of the "7 A peak" in the EXAFS spectra of [Fe(btzp)3](ClO4)2, in contrast with what has been observed for the [Fe(4-R-1,2,4-triazole)3]-(anion)2 chain compounds, confirms that this peak can be used as the signature of a metal alignment only when it involves a strongly enhanced multiple scattering M-M-M path, with M-M spacing less than 4 A. Irradiation with green light at 5 K has led to the population of the metastable high-spin state for the iron(II) ion. The nature of the spin crossover behavior has been discussed on the basis of the structural features. PMID- 11428108 TI - Inversion of the cis geometry requirement for cytotoxicity in structurally novel platinum(II) complexes containing the bidentate N,O-donor pyridin-2-yl-acetate. AB - Water soluble platinum(II) complexes have been synthesized that contain the N,O chelate pyridin-2-yl acetate (PyAc) as a novel structural motif in platinum antitumor complexes. The trans-platinum complex trans-[PtCl(PyAc-N,O)(NH3)] (2) (N-donors are trans) and its isomer cis-[PtCl(PyAc-N,O)(NH3)] (4) (N trans to Cl) were prepared from trans-[PtCl2((NH3)(PyAcH)].H2O (1.H2O) and cis [PtCl2(NH3)(PyAcMe) (3), respectively, employing the bidentate ligand as its methylester (PyAcMe). 2 and 4 are readily formed from the respective dichloro species, even at low pH and in the presence of extra chloride, indicating a high thermodynamic stability of the PyAc chelate ring. 1.H2O and 2-4 were characterized by 1H NMR and IR spectroscopy and elemental analyses. The solid state structure of 2 was determined: triclinic, P1 (no. 2), with a = 8.170(2) A, b = 9.274(3) A, c = 7.374(2) A, alpha = 108.68(2) degrees, beta = 113.27(2) degrees, gamma = 74.40(2) degrees, V = 479.7(6) A3, Z = 2. The six-membered metallacyclus in 2 adopts a "boat" form, allowing a strainless coordination of platinum. The most promising cytotoxic properties in the above series of compounds have been established for 2 (and 1, which rapidly transforms into 2 at 37 degrees C and neutral pH). Preliminary ID50 values were 0.88 and 1.26 microM, respectively, in cisplatin-sensitive L1210 leukemia. Both compounds proved to be cross-resistant to the clinical drug. Reactions of 2 and 4 with 5'-guanosine monophosphate (5'-GMP) under physiological conditions gave the monofunctional adducts trans- and cis-[Pt(5'-GMP-N7)(PyAc-N,O)(NH3)] (I and II). Chelate-bound carboxylate was not replaced by guanine-N7 when an excess of nucleotide was applied (NMR). In an analogous reaction, 2 reacts with the oligonucleotide d(TCGT) [5'-T(1)-C(2)-G(3)-T(4)-3'] to give the adduct d(TCGT)-N7(3)-Pt(PyAc O,N)(NH3) (III), which was characterized by a combination of total correlation spectroscopy, double-quantum-filtered correlation spectroscopy, nuclear Overhauser effect spectrometry, and rotating-frame Overhauser enhancement spectroscopy experiments. Binding of the [Pt(PyAc-N,O)(NH3)]+ fragment to N7 of G(3) causes an increase of N-type character of the T(4) and G(3) deoxyribose residues relative to the unplatinated sequence, while those of T(1) and C(2) remain S-type. An internucleotide nuclear Overhauser effect between H6(4) and H2'(3) indicates stacking between guanine and the 3'-thymine base. The most striking feature proved to be the pronounced upfield shift and broadening of the 1H NMR signals assigned to the base protons H5 and H6 in III. Magnetization transfer between H5(2) and H3 of pyridine suggests that this effect is caused by base-base interactions involving the planar ligand on platinum, which must be situated on the 5' face of guanine. Possible implications for the DNA binding and cytotoxic effect of the compounds are discussed. PMID- 11428110 TI - The synthesis and characterization of bis-substituted derivatives of the [a2 B20H18]4- anion and the interconversion of isomers. AB - The apical-apical (a2) isomer of [Et4N]4[B20H18] reacts with oxalyl chloride in dichloromethane to produce a protonated bis-substituted carbonyl species, [Et4N][a2-B20H17(CO)2] ([Et4N][H1]), in 60% yield. Removal of the bridging hydrogen of the [H1]- anion in aprotic media results in rearrangement to form the equatorial-equatorial [e2-B20H16(CO)2]2- anion ([e2-1]2-). The reaction of [Et4N][H1] with sodium azide in acetonitrile produces [Et4N]3- [a2-B20H17(NCO)2] ([Et4N]3[H2]) in 53% yield, which subsequently reacts with isopropylamine in acetonitrile to provide the urea derivative [a2-B20H16(NH2C(O)NH(i-Pr))2]2- ([Et4N]2[4]) in 89% yield. The [H1]- ion is hydrolyzed in aqueous acetonitrile to give a protonated [a2-B20H17(CO2H)2]3- ion ([H3]3-) in 61% yield. The a2 isomers of the bis-substituted species [B20H16(CO2H)2]4- ([3]4-), [B20H16(NCO)2]4- ([2]4 ), and [B20H16(NH2C(O)NH(i-Pr))2]2- ([4]2-), formed by the removal of the bridging proton from their protonated precursors, rearrange to form a mixture of ae isomers in solution. PMID- 11428111 TI - Triribbed-functionalized clathrochelate iron(II) dioximates as a new and promising tool to obtain polynucleating and polynuclear compounds with improved properties. AB - Template condensation on iron(II) ion of dichloroglyoxime (H2C12Gm) with (C6H5O)3, n-C4H9B(O-n-C4H9)2, and BF3.O(C2H5)2 in CH3NO2 afforded reactive clathrochelate precursors Fe(C12Gm)3(BC6H5)2 (2), Fe(C12Gm)3(B-n-C4H9)2 (3), and Fe(C12Gm)3(BF)2 (4). A series of triribbed-functionalized clathrochelate dioximates have been synthesized. Reaction of 2 with C6H5SH/K2CO3 and CH3SH/t C4H9OK in 1,4-dioxane and THF afforded Fe((C6H5S)2Gm)3(BC6H5)2 (5) and Fe((CH3S)2Gm)3(BC6H5)2 (6). Reaction of 3 with C6H5OK in THF afforded Fe((C6H5O)2Gm)3(B-n-C4H9)2 (7). Condensation of 3 with bis(2-(o oxyphenoxy))diethyl ether in THF afforded di- and tricrown etheric Fe(CwGm)2(C12Gm)(Bn-C4H9)2 (8) and Fe(CwGm)3(B-n-C4H9)2 (9) clathrochelates. Condensation of 3 with 3,5-dithiaoctane-1,8-dithiol/Cs2CO3 in DMF afforded the thiocrown etheric Fe((12anS4)Gm)3(BC6H5)2 complex (10). Reaction of 2 with n butylamine in DMF resulted in the tetrasubstituted Fe((n C4H9NH)2Gm)2(C12Gm)(BC6H5)2 clathrochelate (11). The clathrochelates obtained have been characterized both on the basis of elemental analysis, FAB and PD mass spectrometry, IR, UV-vis, 57Fe Mossbauer, and NMR spectroscopies and crystallographically (compounds 3, 4, 6, 7, and 11). An encapsulated iron(II) ion in a distorted trigonal-prismatic environment of six nitrogen atoms of the macrobicyclic ligand was found to be in a low-spin state. The cyclic voltammograms for the complexes 2-11 show irreversible oxidation waves assignable to Fe3+/Fe2+ couples. The correlation of E1/2 for these couples with Hammet sigma para constants for substituents in dioximate fragments has been demonstrated. PMID- 11428112 TI - Structural distortions in mer-M(H)3(NO)L2 (M = Ru, Os) and their influence on intramolecular fluxionality and quantum exchange coupling. AB - Molecules of the type mer-M(H)3(NO)L2 [M = Ru (1), Os (2); L = PR3] are characterized on the basis of 1H NMR T1min values and IR spectra as pseudo octahedral trihydrides significantly distorted by compression of the cis H-M-H angles to approximately 75 degrees. The distortion, uncharacteristic of six coordinate d6 complexes, is rationalized with DFT (B3LYP) calculations as being driven by increased H-to-M sigma donation and by the exceptional pi-accepting ability of linear NO+. In both 1 and 2, hydrides undergo intramolecular site exchange with delta HHH++(1) = 10-11 kcal/mol and delta HHH++(2) = 16-20 kcal/mol, depending on L, whereas for mer-Ru(H)3(NO)(PtBu2Me)2 (1b), moderate exchange couplings (up to 77 Hz) are featured in the low-temperature 1H NMR spectra, in addition to chemical exchange. On the basis of experimental and theoretical results, a dihydrogen intermediate is suggested to mediate hydride site exchange in 1. The cis H-M-H distortion shortens the tunneling path for the exchanging hydrides in 1, thereby increasing the tunneling rate; diminishes the "conflict" between trans hydrides in the mer geometry; and decreases the nucleophilicity of the hydrides. The generality of the observed structural distortion and its dependence on the ligand environment in late transition metal tri- and dihydrides are discussed. A less reducing metal center is generally characterized by greater distortion. PMID- 11428113 TI - Syntheses, molecular structures, and vibrational spectra of chloropentacarbonylrhodium(III) and -iridium(III) undecafluorodiantimonate(V), [Rh(CO)5Cl][Sb2F11]2 and [Ir(CO)5Cl][Sb2F11]2: an experimental and density functional study. AB - The reactions of either bis(mu-chloro)tetracarbonyldirhodium(I), [Rh(CO)2(mu Cl)]2, or chlorotricarbonyliridium(I), [Ir(CO)3Cl]n, in the conjugate Bronsted Lewis superacid HF-SbF5 and in a CO atmosphere, produce [Rh(CO)5Cl][Sb2F11]2 or [Ir(CO)5Cl][Sb2F11]2, respectively. In these oxidative carbonylation reactions, antimony(V) fluoride functions as an oxidizing agent. The reduced product is identified as 6SbF3.5SbF5. [Rh(CO)5Cl][Sb2F11]2 is obtained in the form of single crystals. Crystal data: monoclinic, space group P2(1) (No. 4); a = 9.721(1), b = 12.602(1), c = 10.538(1) A; beta = 106.51(1) degrees; V = 1237.7(2) A3; Z = 2; T = 300 K; R1 [I > 3 sigma (I)] = 0.0367, wR2 = 0.0739. Single crystals of [Ir(CO)5Cl][Sb2F11]2 are produced in small amounts from a solution of mer Ir(CO)3(SO3F)3 in magic acid, HSO3F-SbF5. The possible source of chlorine will be discussed. Crystal data for [Ir-(CO)5Cl][Sb2F11]2: monoclinic, space group P2(1) (No. 4); a = 9.686(2), b = 12.585(2), c = 10.499(2) A; beta = 106.59(2) degrees; V = 1226.5(4) A3; Z = 2; T = 294 K; R1[I > 3 sigma (I)] = 0.032, Rw = 0.031. The bond lengths and bond angles are nearly identical in the two isostructural salts; however, the cell volume of [Ir(CO)5Cl][Sb2F11]2 is slightly smaller than that of [Rh(CO)5Cl][Sb2F11]2. The cations (point group C4v) feature unusually long M-C bonds (M = Rh, Ir) and correspondingly short CO bonds, as well as high CO stretching wavenumbers and high CO stretching force constants. The [Sb2F11]- anions are not symmetry related, and their dihedral and bridge angles differ slightly in both salts. There are significant interionic contacts in [Ir(CO)5Cl][Sb2F11]2 exclusively of the C-F type (about 2 for each C atom of the five carbonyl groups) resulting in extended structures. The vibrational spectra for both [M(CO)5Cl]2+ cations (M = Rh, Ir) are assigned with the help of density functional calculations, which also provide intensities for IR and Raman bands. While [Rh(CO)5Cl]2+ is the first cationic carbonyl derivative of Rh(III), the vibrational and structural parameters for [Ir(CO)5Cl]2+ are compared to data for [Ir(CO)6]3+ and mer-Ir(CO)3(SO3F)3. PMID- 11428114 TI - Metal-containing ligands for mixed-metal polymers: novel Cu(II)-Ag(I) mixed-metal coordination polymers generated from [Cu(2-methylpyrazine-5 carboxylate)2(H2O)].3H2O and silver(I) salts. AB - One Cu(II)-containing ligand and two Cu(II)-Ag(I) mixed-metal coordination polymers have been synthesized. [Cu(2-methylpyrazine-5-carboxylate)2(H2O)].3H2O (1) was obtained as a molecular complex with two uncoordinated nitrogen donors by the reaction of 2-methylpyrazine-5-carboxylate sodium with CuCl(2).2H2O in water. Compound 1 crystallized in the triclinic space group P1, with a = 10.498(2) A, b = 11.000(2) A, c = 8.1424(16) A, alpha = 98.33(3) degrees, beta = 101.83(3) degrees, gamma = 66.68(3) degrees, and Z = 2. Reactions of 1 with silver(I) salts have been studied. Two Cu(II)-Ag(I) mixed-metal coordination polymers, namely, Ag[Cu(2-methylpyrazine-5-carboxylate)2.(H2O)2](BF4) (2) and Ag[Cu(2 methylpyrazine-5-carboxylate)2.(H2O)2](NO3) (3), have been generated by treating 1 with AgBF4 and AgNO3, respectively. Compound 2 crystallized in the monoclinic space group C2/c, with a = 25.827(5) A, b = 9.6430(19) A, c = 7.4525(15) A, beta = 94.74(3) degrees, and Z = 4. Compound 3 also crystallized in the monoclinic space group C2/c, with a = 25.855(5) A, b = 9.782(2) A, c = 7.1201(14) A, beta = 96.90(3) degrees, and Z = 4. The main structural feature in both 2 and 3 is a zigzag Cu(II)-Ag(I) mixed-metal chain, in which the alternating Cu(II) and Ag(I) centers are linked by 2-methylpyrazine-5-carboxylate spacers. The effect of the nitrate counterion was illustrated by compound 3, in which a novel [Ag+...NO3-] coordination chain has been found which acts as the connector to cross-link the one-dimensional zigzag chains into a three-dimensional network. In addition, an identical interchain O-H...O hydrogen bonding system has been found in both 2 and 3 and has been shown to play a significant role in directing the alignment of the one-dimensional mixed-metal polymer chains in the crystalline state. The magnetic susceptibilities of 2 and 3 were measured and found to follow the Curie law (mu eff = 1.85 for 2 and 1.83 for 3). PMID- 11428115 TI - Kinetics and mechanism of complex formation reactions in the Iron(III)-phosphate ion system at large iron(III) excess. Formation of a tetranuclear complex. AB - The kinetics and mechanism of the iron(III)-phosphate ion reaction were studied at large iron(III) excess using the stopped-flow method at 10.0 degrees C in 1.0 M NaClO4. In the first few hundred milliseconds of the reaction, the formation of a novel tetranuclear complex was confirmed. The following composition is proposed for the new species: Fe4(PO4)(OH)2(H2O)16(7+). According to detailed kinetic studies, the formation of this species is first order with respect to Fe2(OH)2(H2O)8(4+) and H2PO4- and presumably proceeds via a dinuclear intermediate species. At longer reaction times slow dissociation of the tetranuclear complex controls the formation of the thermodynamically favored Fe(PO4)(H2O)5 complex. The overall reaction was interpreted in terms of the following reactions: Fe2(OH)2(H2O)8(4+) [symbol: see text] 2Fe3+ mn; Fe2(OH)2(H2O)8(4+) + P(V) [symbol: see text] Fe2PV; Fe2PV + Fe2(OH)2(H2O)8(4+) [symbol: see text] Fe4PV; Fe3+ mn + P(V) [symbol: see text] Fe(PO4)(H2O)5. (Fe3+ mn = Fe(H2O)6(3+) + Fe(OH)(H2O)5(2+); P(V) = H3PO4 + H2PO4-; Fe2PV = Fe2(HPO4)(OH)(H2O)8(3+); Fe4PV = Fe4(PO4)(OH)2(H2O)16(7+).) The pH dependence and relevant rate and equilibrium constants are reported for the individual reaction steps. PMID- 11428116 TI - Chloroaluminate ionic liquids as reagents for isolating soluble hexanuclear zirconium halide cluster compounds. AB - Ambient-temperature chloroaluminate molten salts, mixtures of aluminum trichloride (AlCl3) and 1-ethyl-3-methylimidazolium chloride (ImCl), have been used as solvents to excise and isolate centered hexanuclear zirconium halide clusters from their solid-state precursors. Cluster compounds synthesized via high-temperature reactions, KZr6CCl15 and Li2Zr6MnCl15, were dissolved into basic molten salts at 100-110 degrees C. The C-centered cluster compound, Im4Zr6CCl18, was isolated in 70% yield, and the Mn-centered cluster compound, Im5Zr6MnCl18.C7H(8).2CH3CN, was isolated in 54% yield. Im5Zr6BCl18 is efficiently oxidized by ferrocenium tetrafluoroborate, and one-electron-oxidized B-centered cluster, [(Zr6B)Cl18]4-, was isolated in 90% yield as the salt Im4Zr6BCl18. PMID- 11428117 TI - Kinetics and mechanisms of the oxidation of hydrazinium ion (N2H5+) by aqueous Br2, Cl2, and BrCl. Electrophilicity scale for halogens and interhalogens. AB - Very rapid oxidations of N2H5+ by Br2, Cl2, and BrCl are measured by stopped-flow and pulsed-accelerated-flow methods in acidic solutions with excess N2H5+. Second order rate constants (M-1 s-1) at 25.0 degrees C, mu = 1.0 M are 1.49 x 10(7), 1.01 x 10(8), and 5.6 x 10(8) for the reactions with Br2, Cl2, and BrCl, respectively. The reactions are postulated to proceed by nucleophilic reaction of N2H5+ with XY electrophiles (XY = Br2, Cl2, BrCl) to form XN2H4+ with Y- and H+ release in the rate-determining step. In the subsequent reactions, we propose that XN2H4+ eliminates X- and H+ rapidly to form N2H3+ and diazine, N2H2, which is oxidized by a second Br2, Cl2, or BrCl to form N2 in fast steps. The stoichiometries are measured and confirmed to be 1:2 for the Cl2 and BrCl reactions. The relative reactivities for the oxidation of N2H5+ by halogens and interhalogens (BrCl > Cl2 > Br2 > ICl >> IBr >> I2) are used to establish an electrophilicity scale (EXY) for this type of reaction in aqueous solution. PMID- 11428118 TI - Spin-state variation in solid state and solution of mononuclear iron(II) 1,4,7 trimethyl-1,4,7-triazacyclonane complexes. AB - The series of mononuclear iron(II) complexes with the tridentate macrocycle Me3tacn have been prepared (Me3tacn = 1,4,7-trimethyl-1,4,7-triazacyclonane). A purple, spin-crossover complex [Fe(Me3tacn)(MeCN)3](CF3-SO3)2 (1-OTf) forms in acetonitrile but readily loses MeCN ligands to form a colorless high-spin complex Fe(Me3tacn)(OTf)2 (2). The BPh4- salt of 1 is stable to loss of MeCN and remains purple even under a vacuum. Methylene chloride solutions of Fe(OTf)2 and Me3tacn afford crystals of [Fe(Me3tacn)(MeCN)2(OTf)](OTf) (3). Crystallization of 1-OTf in the presence of water affords a colorless high-spin complex, Fe(Me3tacn)(H2O)(CF3-SO3)2 (4), that exists as a pair of molecules bridged by hydrogen bonds between the coordinated water and the two bound triflate anions of the inversion-related partner. The crystallographic parameters are the following. 1-BPh4: C63H70B2Fe, monoclinic, P2(1)/c, a = 18.360(1) A, b = 11.761(1) A, c = 25.754(2) A, beta = 90.72(1) degrees, Z = 4. 3: C16H29Cl2F6FeN5O6S2, triclinic group P1, a = 8.500(1) A, b = 11.421(2) A, c = 15.677(2) A, alpha = 92.23(1) degrees, beta = 94.79(1) degrees, gamma = 97.03(1) degrees, Z = 2. 4: C20H18F6FeN4O6S2, monoclinic, P2(1)/n, a = 11.253(3) A, b = 12.624(5) A, c = 14.683(5) A, beta = 94.02(2) degrees, Z = 4. Variable temperature visible spectra and 1H NMR spectra of solutions of both 1-OTf and 1-BPh4 exhibit low-spin, high spin crossover behavior, whereas 2, 3, and 4 remain high-spin in solution. The extensive role of coordinated triflate as a terminal and/or bridging ligand as well as a counteranion is demonstrated by variable temperature 19F NMR spectra. PMID- 11428119 TI - Coordination polymers: structural transformation from two to three dimensions through ligand conformation change. PMID- 11428120 TI - EPR characterization of the products formed after photolysis of [CoIII(Salen)(CH3)(H2O)] and [CoIII(SaltMe)(CH3)(H2O)] in the presence of N- and P-donor bases. PMID- 11428121 TI - SiONB unit as reference for blocked Si...N interactions in SiON compounds. PMID- 11428122 TI - Titanium and niobium imido complexes derived from diamidoamine ligands. PMID- 11428123 TI - A decairon cluster devoid of polydentate ligands. PMID- 11428124 TI - Synthesis and characterization of (hfac)In(CH3)2: a volatile compound useful for CVD of indium and indium-containing materials. PMID- 11428125 TI - Low-melting, mononuclear tetrahydrofuran complexes of M(2,2,6,6 tetramethylheptane-3,5-dionate)2 (M = Ba, Sr) and related analogues. PMID- 11428126 TI - Clinical significance of weight loss in cancer patients: rationale for the use of anabolic agents in the treatment of cancer-related cachexia. PMID- 11428127 TI - Thomas J. King Jr. 1921-2000. PMID- 11428128 TI - Two B or not two B? Overview of the rapidly expanding B-box family of proteins. AB - The B-box gene family represents a large number of genes involved in functions such as axial patterning, growth control, differentiation, and transcriptional regulation. These genes possess several conserved motifs that always include a B box zinc binding motif associated with various other motifs such as the RING zinc finger, an alpha-helical coiled-coil, the rfp or B30.2 motif, propeller domain, and the NHL motif in various combinations. Mutations or rearrangements in several B-box family members are associated with human diseases and cancers such as familial Mediterranean fever (FMF), Optiz/BBB syndrome, acute promyelocytic leukemia, mulibrey nanism, and thyroid carcinomas. This suggests that members of this gene family play important roles in fundamental biological processes. Here we discuss the known members of this rapidly expanding protein family. PMID- 11428129 TI - Spore coat formation and timely sporulation depend on cellulose in Dictyostelium. AB - Cellulose is a major component of the extracellular coat that surrounds the terminally-differentiated spore of Dictyostelium. It is sandwiched between two layers of proteins that derive from prespore vesicles by exocytosis. Strains unable to synthesize cellulose due to null mutations in the gene encoding the catalytic subunit of cellulose synthase (dcsA) failed to make detergent-resistant spores but produced small, highly refractile, round spore-like cells up to a day late. Although these cells resembled spores in appearance, they were unstable, only transiently ellipsoid in shape, and sensitive to hypo-osmotic shock, drying, or detergents. Differentiation of these pseudo-spores was induced in the normal time frame by activation of the cAMP-dependent protein kinase or co-development with wild type cells, and coat proteins were secreted by the dcsA-null cells at the same time as wild type cells. A substantial fraction of secreted coat proteins was loosely associated with the surface of the mutant cells, resembling the precoat posited to form early during normal sporulation. Transmission electron microscopy revealed that the precoat had little ultrastructural organization in the absence of cellulose. Thus, cellulose in the coat appears to be required for the organization of the pre-coat precursors as well as the stability, dormancy, and shape of the spore. PMID- 11428130 TI - Mitochondrial ribosomal RNA in the germinal granules in Xenopus embryos revisited. AB - Germ cells of various animals contain a determinant that is called the germ plasm. In amphibians such as Xenopus laevis, the germ plasm is composed of mitochondria and electron dense germinal granules that are embedded in a fibrillar matrix. Previous reports indicated that one of the components of germinal granules was mitochondrial large and small ribosomal RNA (mtlrRNA and mtsrRNA). Utilizing a modified procedure for electron microscopy in situ hybridization, we investigated the distribution of these RNAs along with other components of the germ plasm in Xenopus laevis embryos. We found, that contrary to previous reports, the mtlrRNA and mtsrRNA were located in close vicinity to the germinal granules but were not major constituents of granules. The majority of the mtlrRNA and mtlsrRNAs was present inside the mitochondria and in the germ plasm matrix. PMID- 11428131 TI - Gli2 mediation of hedgehog signals in slow muscle induction in zebrafish. AB - Zebrafish skeletal muscles are composed of two major types of muscle fibers, broadly classified as fast or slow fibers. Recent studies have demonstrated that members of the Hedgehog (Hh) family induce the formation of slow muscle fibers. Hedgehog signals are secreted proteins that function through the transcription factor Glis. We report here the characterization of a zebrafish Gli2 expression in slow and fast muscle cells and the study of the roles of Hedgehogs and Gli2 in zebrafish muscle development using two mutant strains; sonic-you (syu) and you too (yot), respective for sonic hedgehog (shh) and Gli2 mutation. We have demonstrated that Shh and Gli2 mutation causes similar defects in slow muscle formation. There is, however, a difference in the degree of defect between these two mutants. In yot mutant embryos, development of slow muscles was completely blocked, whereas in syu mutant embryos, a small number of slow muscle cells could still form, suggesting that other Hhs were also involved in slow muscle induction. Induction of slow muscles by other Hhs appeared to require Gli2, because ectopic expression of Echidna hedgehog (Ehh) and Tiggy-winkle hedgehog (Twhh) failed to induce slow muscles in yot mutant embryos. Together, these data suggest that further Hhs, other than Shh, are also involved in the induction and differentiation of slow muscle cells and that Gli2 is required by Shh, Twhh, and Ehh, thus playing a key role in the induction and differentiation of slow muscle cells. PMID- 11428132 TI - Effect of chronic ozone fumigation on the photosynthetic process of poplar clones showing different sensitivity. AB - Rooted cuttings from two poplar clones (Populus x euramericana, I-214, and Populus deltoides x maximowiczii, Eridano) were exposed for 15 days to diurnal square-wave treatment with ozone (60 nL L-1 for 5 h day-1). Completely fully expanded leaves exposed to ozone showed a reduction in net CO2 assimilation rate as compared to the control leaves during whole exposure period in both the clones. The reduction was related to a strong stomatal closure in clone I-214, but also to an altered mesophyll activity ascribed to limitation of the dark reactions of photosynthetic process. The results obtained in leaves of I-214 subjected to long-term fumigation seem to support the view that the decrease in quantum yield of electron transport may be a mechanism to down-regulate photosynthetic electron transport so that production of ATP and NADPH would be in equilibrium with the decreased demand in the Calvin cycle. In Eridano the CO2 assimilation was reduced because of the exposure and any alteration in stomatal conductance was observed. Thus, chlorophyll fluorescence parameters showed that an inhibition of photosystem II had occurred (reduction in Fv/Fm ratio), while no alterations in quenching parameters were observed upon illumination. The results seem to indicate that an alternative sink for reducing equivalent, other than carbon metabolism is present. PMID- 11428133 TI - Hydrocarbon deposition and soil microflora as affected by highway traffic. AB - The proximity of a busy highway (90,000 vehicles/day) increased the amount of polycyclic aromatic hydrocarbons (PAHs) in soil at the depth of 5-15 cm from 106 ng/g as a grassland background to 3095 ng/g dry soil at the highway verge (a sum of 10 PAH species). The PAH concentration was related to the distance from the source and exhibited a biphasic character, which is interpreted in terms of bimodal distribution of the exhaust microparticles with different rates of deposition. Similarly, the tendency of benz(a)anthracene, chrysene, benzo(b)fluoranthene, benzo(k)fluoranthene, and indeno(1,2,3-cd)pyrene to decrease their proportion with distance from the highway, in contrast to phenanthrene, fluoranthene, pyrene, benzo(a)pyrene, and benzo(g,h,i)perylene, was attributed to their prevalent localisation on the heavier particle fraction. The abundance of bacteria (8.33 x background) and fungi (3.17 x background) close to the highway is thought to be a consequence of hydrocarbon deposition from the traffic that serves as a significant energetic input into the soil. The elevated concentrations of hydrocarbon substrates, as indicated by PAHs, increased both the absolute and relative numbers of the microbial degraders of diesel fuel, biphenyl, naphthalene, and pyrene. Their maximum numbers at 0.5-1.5 m from the pavement reached 1.3 x 10(4), 1.2 x 10(5), 1.1 x 10(4), and 6.6 x 10(3) colony forming units (CFU) or infection units per gramme dry soil, respectively. On the other hand, the number of anthracene degraders (1.1 x 10(3) CFU per g dry soil) remained close to the detection limit of the enumeration technique used (0.1-0.2 x 10(3) per g dry soil), consistently with the absence of anthracene and higher linear PAHs in the investigated soil samples. The amounts of persisting PAHs justify artificial inoculation with effective degrader strains in the vicinity of motorways. PMID- 11428134 TI - Extraction of chlorinated aliphatic hydrocarbons from groundwater at micromolar concentrations for isotopic analysis of chlorine. AB - A method is described for near-quantitative extraction of micromolar concentrations of chlorinated aliphatic hydrocarbons (CAHs) from water for determination of chlorine (Cl) isotope ratios. A low pressure, carrier-gas procedure of extraction was proven to be applicable to CH2Cl2, CCl4, C2H2Cl2, and C2HCl3. The pH of the water was adjusted with NaOH to prevent extraction of CO2 from air and/or dissolved inorganic carbonate species. Recoveries of CAH samples (approximately 15 mumol), added to and extracted from approximately 340 ml of water, averaged approximately 96%. Average changes in the delta 37Cl values of the CAHs, attributable to the extraction process, were -0.01 +/- 0.06@1000. Significant isotopic fractionation of Cl was measured during partial extraction of C2CHCl3 from water, indicating that near-quantitative extraction is required for reliable stable Cl isotope analysis of CAHs. This method is also suitable for the extraction of dissolved CAH for gas chromatography-combustion-isotope ratio mass spectrometric measurements of hydrogen and carbon. PMID- 11428135 TI - Effects of oxidants on soybean growth and yield in the Pakistan Punjab. AB - Plants of soybean (Glycine max L.) were grown with and without the ozone protectant EDU (N-[2-(2-oxo-1-imidazolidinyl)ethyl]-n2 phenylurea) at a suburban site, a remote rural site and a rural roadside site around the city of Lahore, Pakistan. The development and yield of the plants was determined in two experiments--one immediately post-monsoon and one in the following spring (pre monsoon). Concentrations of nitrogen dioxide and photochemical oxidants were measured at each site. The effect on yield of EDU at the suburban site (47 and 113% increase in seed weight per plant relative to the untreated plants in the post- and pre-monsoon experiments, respectively) was similar to the effects of filtration on yield on soybean in a parallel open-top chamber study at the same site (77% increase relative to plants subjected to unfiltered air for the pre monsoon experiment). Effects of EDU on yield were greater at both rural sites than at the suburban site in both experiments, and greater in the spring experiment (182% at the remote rural site and 285% at the rural roadside site) than in the post-monsoon experiment (94% at the remote rural site and 170% at the rural roadside site); oxidant concentrations were also greater at the rural sites than at the suburban site, and greater in the spring experiment than the post monsoon experiment. The results imply that ozone may be causing significant crop losses in rural areas around Lahore; however, the geographical extent of the problem, and the implications for peri-urban agriculture around other cities of south Asia are uncertain. PMID- 11428136 TI - Development of a Marine Sediment Pollution Index. AB - To facilitate translation of the state of marine sediment quality for public information, a pollution index was developed from the results of a routine monitoring program. Principal component analysis (PCA) of 24 variables at 64 monitoring stations from 1987 to 1997 was carried out to identify the most important parameters that were applied in the index formulation. Of the 24 variables, six (Cr, Cu, Ni, chemical oxygen demand, Zn, Mn) were selected on the basis of their high PCA loadings. The derived Marine Sediment Pollution Index, rating from 0 to 100, reflected the general trend in the monitoring areas. The index was also found to have a significant negative correlation (P < 0.05) with the benthic species diversity and toxicity of the sediment, indicating its usefulness in reflecting marine sediment quality. The application of PCA to identify important variables from a monitoring program would reduce sampling resources, as parameters that did not show significant spatial or temporal variations could be analyzed in a lesser frequency than those that were identified to be more important from the results of PCA. PMID- 11428137 TI - Relationship of properties of polycyclic aromatic hydrocarbons to sequestration in soil. AB - A study was conducted to determine whether the sequestration of 21 polycyclic aromatic hydrocarbons (PAHs) in soil was correlated with their properties. From 22 to 58% of the PAHs was not extracted with n-butanol after their addition to soil. After 28 days of aging, the percentage of the PAHs remaining in the soil increased to 47-77%; however, nearly all of each compound was recovered by Soxhlet extraction. Correlations were based on the amounts of aged compound extracted with butanol. Properties of compounds used in the correlations included Kow, molecular length and molecular-connectivity indices (MCIs). No one property, including log Kow, resulted in an R2 value greater than 0.26. A chain MCI (2 chi vCH) together with log Kow or a first-order MCI (1 chi) resulted in R2 values of 0.49 and 0.54, respectively. The data suggest that the properties tested are not important to predicting the sequestration of PAHs in soil. PMID- 11428138 TI - Use of radium isotopes to determine the age and origin of radioactive barite at oil-field production sites. AB - Radium-bearing barite (radiobarite) is a common constituent of scale and sludge deposits that form in oil-field production equipment. The barite forms as a precipitate from radium-bearing, saline formation water that is pumped to the surface along with oil. Radioactivity levels in some oil-field equipment and in soils contaminated by scale and sludge can be sufficiently high to pose a potential health threat. Accurate determinations of radium isotopes (226Ra + 228Ra) in soils are required to establish the level of soil contamination and the volume of soil that may exceed regulatory limits for total radium content. In this study the radium isotopic data are used to provide estimates of the age of formation of the radiobarite contaminant. Age estimates require that highly insoluble radiobarite approximates a chemically closed system from the time of its formation. Age estimates are based on the decay of short-lived 228Ra (half life = 5.76 years) compared to 226Ra (half-life = 1600 years). Present activity ratios of 228Ra/226Ra in radiobarite-rich scale or highly contaminated soil are compared to initial ratios at the time of radiobarite precipitation. Initial ratios are estimated by measurements of saline water or recent barite precipitates at the site or by considering a range of probable initial ratios based on reported values in modern oil-field brines. At sites that contain two distinct radiobarite sources of different age, the soils containing mixtures of sources can be identified, and mixing proportions quantified using radium concentration and isotopic data. These uses of radium isotope data provide more description of contamination history and can possibly address liability issues. PMID- 11428139 TI - Effects of oil sands effluent on cattail and clover: photosynthesis and the level of stress proteins. AB - The oil sands industry located in northeastern Alberta, Canada, generates large volumes of effluent characterized by a high level of dissolved ions and naphthenic acids. The dikes used to store the effluent seep, creating wetlands which are subsequently invaded by obligate wetland flora such as cattail (Typha latifolia L.). The appearance of these wetlands prompted the oil sands industry to consider wetlands as part of their reclamation strategy. However, to ensure long-term viability of such wetlands, the response of the flora to the industrial effluent needed to be determined. To this end, apparent photosynthesis (APS), the level of ribulose-1,5-bisphosphate carboxylase (RuBisCo) large subunit, dehydrin related polypeptides, and protein disulphide isomerase (PDI) were evaluated in cattail and alsike clover plants (Trifolium hybridum L.) exposed to the oil sands effluent. APS measured in plants impacted by oil sands effluent was significantly higher than that of plants in the non-impacted off-site location. Among the on site locations, plants growing in the natural wetlands site had higher APS compared to all other sites. The level of RuBisCo was not increased in cattail or clover growing in effluent-contaminated sites indicating that enhanced photosynthesis was not due to greater levels of this enzyme. Dehydrin-related polypeptides were detected only in the roots of cattail and were absent in clover. The polypeptide profile was altered in cattail exposed to oil sands effluent indicating that they were responding to an osmotic stress. The level of PDI was unaffected in the leaves of cattail regardless of the nature of the effluent to which they were exposed. Overall, the data indicate that cattail and clover are adapted to the oil sands effluent, although further studies are needed to assess their long-term ability to survive in the presence of this anthropogenic stress. PMID- 11428140 TI - Efficacy of histopathology in detecting petrochemical-induced toxicity in wild cotton rats (Sigmodon hispidus). AB - A variety of chemical mixtures exist in the soil of petrochemical waste sites, and many of these compounds are known immunotoxicants that have been observed to induce immune alterations in wild rodents inhabiting many of these petrochemical waste sites. Conventional histopathological assessments have been widely used with considerable success to investigate immunotoxicity of various agents under laboratory conditions. We hypothesized that histopathologic assessments would be equally sensitive for detecting exposure to complex mixtures of toxicants in cotton rats (Sigmodon hispidus) residing in contaminated habitats. Histopathological parameters were examined from a total of 624 cotton rats that were seasonally collected from 13 petrochemical-contaminated waste sites and 13 ecologically matched reference sites in Oklahoma over a 3-year period. Histopathological examination did not reveal any lesion associated with exposure to petrochemical wastes except renal inclusion bodies. Prevalence and severity of histologic lesions in liver and kidneys of cotton rats were significantly influenced by season, where prevalence and severity were lower in winter than summer on all study sites. These results suggest that the evaluation of toxicity from exposure to contaminants in the soil of industrial waste sites using histopathological assessments is not sensitive enough to detect exposure to the low levels of environmental contaminants present on most waste sites. PMID- 11428141 TI - Characteristics of different molecular weight fractions of organic matter in landfill leachate and their role in soil sorption of heavy metals. AB - We have characterised two kinds of municipal landfill leachates derived from 'old' and 'young' municipal waste landfills on the basis of the molecular weight distribution of the constituents, taking into account that the great variety of leachate constituents prevents any evaluation of the fate and of the role played by each component in the environmental impact. In the sample S1 (old leachate), the constituents were distributed over a wider range of molecular weights; high molecular weight fractions were present. In sample S2 (young leachate), the fractions are actually narrower at the lower molecular weights. The high molecular weight fractions of old leachates are found to be complex structures formed by condensed nuclei of carbons substituted by functional groups containing nitrogen, sulphur and oxygen atoms; the low molecular weight fractions of leachates are, instead, characterised by linear chains substituted by oxygenated functional groups such as carboxyl and/or alcoholic groups. After characterising each fraction we studied the role played by these fractions in the soil's capability for retaining heavy metals [copper (Cu) and cadmium(Cd)]. The Cd uptake increases only on the soil treated with sample S1 characterised by a higher pH value and by the presence of high molecular weight fractions. The Cu uptake also increases on the soil treated with sample S2, characterised by the sole presence of low molecular weight fractions. On the other hand, the metal adsorption tests performed on soil treated with the single fractions show that the amount of Cu and Cd retained by soil treated with the high molecular weight fractions of sample does not increase after 72 h of treatment and that the amount of Cu retained by the low molecular weight fractions of sample S1 and by the fractions of sample S2 increases, but does not justify the amount retained by soil treated with the total leachates. PMID- 11428142 TI - First report on the uptake of automobile catalyst emitted palladium by European eels (Anguilla anguilla) following experimental exposure to road dust. AB - Following the introduction of automobile catalysts in the middle of the 1980s in Germany there is an increasing emission of the platinum-group-metals platinum, palladium (Pd) and rhodium. Still, it remains unclear if these metals are bioavailable for aquatic animals and to which extent they become accumulated by the aquatic biosphere. Because of analytical problems in detecting Pd in small biological samples the present investigation concentrates on the bioavailability of this metal. To answer the question of a Pd uptake by aquatic organisms experimental studies were conducted with European eels maintained in water containing road dust at a concentration of 10 kg/100 l. Following an exposure period of four weeks, samples of liver and kidney were analysed by total reflection X-ray fluorescence analysis after co-precipitation of Pd with mercury. These experiments revealed an uptake of traffic related Pd by European eels which showed a mean liver Pd concentration of 0.18 +/- 0.05 ng/g (wet wt.), whereas the Pd concentration in the kidney ranged below the detection limit. Thus, in this study we can demonstrate for the first time that automobile catalyst emitted Pd is bioavailable for aquatic animals. PMID- 11428143 TI - Metal levels in body tissues, forage and fecal pellets of elk (Cervus elaphus) living near the ore smelters at Sudbury, Ontario. AB - Ontario's only free-ranging elk herd located at Burwash-French River has been subjected to the influence of historic copper-nickel-iron (Cu-Ni-Fe) ore smelting operations within the Sudbury area. Metal levels in selected body tissues, preferred forage items and fecal pellets were determined, and their potential effects on health and reproductive viability assessed. Significant age effects were apparent in 13 out of 31 tissue metal values obtained, with most age-related differences attributable to elevated concentrations in fetal tissues. Cu and zinc (Zn) levels were four to six times higher in livers and one and a half to two times higher in the bones of fetuses as compared to post-natal animals. Enhanced pre-natal tissue burdens were also noted for cobalt (Co), Ni and lead (Pb) in muscles. However, mean cadmium (Cd) levels in kidney, liver and muscle tissues, as well as hepatic Fe concentrations, showed significant increases with age. Although Cd levels in the kidneys of older elk were still well below the reported threshold for irreversible organ damage, and not considered to be health threatening, detailed histological study of the cortical tissues for subtle pathotoxicological effects may be warranted. Unlike renal and hepatic tissues, Cd concentrations in skeletal muscle were minimal and within levels considered acceptable for human consumption. Forage species collected in the elk range generally showed higher (two to eight times) Ni and Fe levels than the corresponding plants from a non-contaminated reference site. The highest Ni and Fe burdens were carried by eastern white cedar, a preferred winter food item for elk. No consistent site-related trends were demonstrated by Cu, Zn, Co, Pb, chromium (Cr) and Cd forage levels. Mean metal concentrations in fecal pellets generally exceeded the mean composite forage levels by a factor of 3, except for Ni, which was five times more concentrated in feces. Significantly elevated Ni and Fe levels in preferred browse species and fecal pellets of the Sudbury-area elk could be of concern and warrant further investigation. PMID- 11428144 TI - Small-scale spatial and temporal variance in the concentration of heavy metals in aquatic sediments: a review and some new concepts. AB - Uncertainty associated with data derived by the analyses of heavy metals in aquatic sediment is due to variance produced in the laboratory (precision), plus 'natural', small-scale spatial variance, (or field variance) at the sampling site. Precision is easily determined and is usually reported in contaminant studies, but field variance is poorly understood and seldom documented. It is important to have an understanding of the field variance because if small-scale spatial variance in the concentration of heavy metals is excessive, regional trends may be limited value. Similarly, if temporal change is large, the results of single synoptic surveys may be questionable and the ability to demonstrate anthropogenic contributions over time will be difficult. However, it is evident from the literature that the information needed to address problems of spatial and temporal variance in the field is beyond the resources of most researchers. Analytical precision of about 5% relative standard deviation (RSD) for heavy metal analysis is typical of a well-managed laboratory. Many studies of small scale spatial variability made during the current investigation indicate that field variance is related to ambient energy and to the type of sedimentological environment. Total variance (analytical plus field variance) is approximately 10% RSD (mean for a suite of nine trace elements) for depositional parts of estuaries and the marine environment, but increases to about 20-35% RSD for the more dynamic parts of the estuarine environment and the fluvial system. Repeated sampling over periods of up to 7 years undertaken during the present study, indicate a similar order of magnitude for temporal variability in these sedimentological environments. A proposed scheme to provide information on field variance is to undertake small-scale spatial and temporal studies in discrete sedimentological environments in the study area after sediment sampling and characterisation has been completed. The comparatively large proportion of total variance associated with small-scale spatial and temporal variability in the field questions the often excessive cost and effort made in attempting minor reductions in analytical precision in contaminant investigations. PMID- 11428145 TI - Polychlorinated biphenyl congeners and organochlorine insecticides in the water column and sediments of Daya Bay, China. AB - Samples of sub-surface water, suspended particulate matter (SPM) and surface sediments collected from Daya Bay, China have been analysed for 12 polychlorinated biphenyl (PCB) congeners and 18 organochlorine insecticides, using gas chromatography electron capture detection. Total PCB levels varied from 91.1 to 1355.3 ng l-1 in water and from 0.85 to 27.37 ng g-1 dry weight in sediments. The levels of total organochlorine insecticides were in the range 143.3-5104.8 ng l-1 in water and 2.43-86.25 ng g-1 dry weight in sediment. None of the target compounds were detected in SPM. The levels of total hexachlorocyclohexanes in water varied from 35.5 to 1228.6 ng l-1, whilst in sediments they ranged from 0.32 to 4.16 ng g-1 dry weight. For the sum of dichlorodiphenyltrichloroethanes (DDTs), their levels were in the range 26.8 975.9 ng l-1 in water, and 0.14-20.27 ng g-1 dry weight in sediments. The distribution profiles of these contaminants in water and sediments suggest that there are a number of sources contributing to total contaminant burden in the bay, including soil runoffs, wastewater discharges, sewage outfalls and shipping activites. Ratios of DDT/(DDE + DDD) in the water and sediments indicate recent inputs of such chemicals into the day. The results, therefore, provide important information on the current contamination status of a key aquacultural area in China, and point to the need for urgent actions to stop the use of persistent agrochemicals such as DDT and lindane. PMID- 11428146 TI - Bioaccumulation of heavy metals in terrestrial invertebrates. AB - In this literature study, accumulation data of metals in terrestrial invertebrates were collected and compared (Arthropoda and Lumbricidae). Based on total soil concentrations and body concentrations, regression equations were calculated for each metal (Cd, Cu, Pb and Zn) and each taxonomic group. We also tried to find out whether or not accumulation levels of metals in Lumbricidae are representative for all of the studied terrestrial invertebrates. Taxonomic groups could be ordered according to the extent of metal accumulation. Significant differences in accumulation levels of a factor 2-12 were found between taxonomic groups. Overall, metal concentrations were high in Isopoda and low in Coleoptera. The concentrations in Lumbricidae were in between. It should be kept in mind that the data for Lumbricidae were mainly derived from laboratory experiments, while the data for other groups were derived from field studies. The internal Pb, Cd and Cu concentration increased with the soil concentration for most taxonomic groups in the order Pb > Cd > Cu. Body concentrations of Zn were quite constant over a range of soil concentrations. The differences in accumulation level between taxonomic groups show the relevance of including detailed information on feeding behaviour in risk assessment for invertebrate-eating animals. PMID- 11428147 TI - Towards the characterisation of heavy metals in dredged canal sediments and an appreciation of 'availability': two examples from the UK. AB - Canal sediments can act as sinks for a wide range of contaminants including heavy metals from various sources (e.g. industrial and waste water discharges). Dredging of canals is required to maintain navigational depth and prevent flooding. The sediments removed from canals are often disposed of to land, being deposited either straight on to the banks of the canal or, in recent years, in licensed disposal sites. The aim of this work was to investigate the nature of dredged sediment-derived soils and the heavy metals present in them. Two disposal sites in the United Kingdom (UK) were investigated and soil samples taken. A variety of analytical techniques were used, including Aqua regia digestion and sequential extraction, in order to assess the concentrations and associations of metals present. Diethylene triaminepenta-acetic acid extracts, performed to illustrate plant-available metal concentrations, reveal that up to 40% of the total extracted metals were in an 'available' form. Variations in metal concentrations with depth in the soil cores show a significant correlation with total organic carbon content. PMID- 11428148 TI - Dialogical nature of cognition. PMID- 11428149 TI - Face-to-face play: its temporal structure as predictor of socioaffective development. PMID- 11428150 TI - Rhythms of dialogue in infancy: coordinated timing in development. AB - Although theories of early social development emphasize the advantage of mother infant rhythmic coupling and bidirectional coordination, empirical demonstrations remain sparse. We therefore test the hypothesis that vocal rhythm coordination at age 4 months predicts attachment and cognition at age 12 months. Partner and site novelty were studied by recording mother-infant, stranger-infant, and mother stranger face-to-face interactions in both home and laboratory sites for 88 4 month-old infants, for a total of 410 recordings. An automated dialogic coding scheme, appropriate to the nonperiodic rhythms of our data, implemented a systems concept of every action as jointly produced by both partners. Adult-infant coordination at age 4 months indeed predicted both outcomes at age 12 months, but midrange degree of mother-infant and stranger-infant coordination was optimal for attachment (Strange Situation), whereas high ("tight") stranger-infant coordination in the lab was optimal for cognition (Bayley Scales). Thus, high coordination can index more or less optimal outcomes, as a function of outcome measure, partner, and site. Bidirectional coordination patterns were salient in both attachment and cognition predictions. Comparison of mother-infant and stranger-infant interactions was particularly informative, suggesting the dynamics of infants' early differentiation from mothers. Stranger and infant showed different patterns of vocal rhythm activity level, were more bidirectional, accounted for 8 times more variance in Bayley scores, predicted attachment just as well as mother and infant, and revealed more varied contingency structures and a wider range of attachment outcomes. To explain why vocal timing measures at age 4 months predict outcomes at age 12 months, our dialogue model was construed as containing procedures for regulating the pragmatics of proto-conversation. The timing patterns of the 4-month-olds were seen as procedural or performance knowledge, and as precursors of various kinesic patterns in the outcomes of 12-month-olds. Thus, our work further defines a fundamental dyadic timing matrix--a system that guides the trajectory of relatedness, informing all relational theories of development. PMID- 11428151 TI - The development and piloting of a capacity assessment tool. PMID- 11428152 TI - Too much ethics, not enough medicine: clarifying the role of clinical expertise for the clinical ethics consultant. PMID- 11428153 TI - How to determine competency. PMID- 11428154 TI - Referral and decision making among advanced cancer patients participating in Phase I trials at a single institution. PMID- 11428155 TI - The use of e-mail in clinical ethics case consultation. PMID- 11428156 TI - Active voluntary euthanasia, terminal sedation, and assisted suicide. PMID- 11428157 TI - Physician-assisted suicide or voluntary euthanasia: a meaningless distinction for practicing physicians? PMID- 11428158 TI - Telemedicine and end-of-life care: what's wrong with this picture? PMID- 11428159 TI - Telethics and the virtual intensivist--a comment on Pronovost and Williams. PMID- 11428160 TI - Should ethics consultants use telemedicine? A comment on Pronovost and Williams. PMID- 11428162 TI - Matrix biology: past, present and future. AB - Matrix biology (the biology of extracellular matrix) is a relatively recent branch of biomedical sciences and comprises a number of subspecialties. From molecular-cell biology, biochemistry, genetics and clinical science of diseases localised at or affecting the matrix rich tissues (connective tissues) as bone, cartilage, vessel wall, skin, eye and some others. The rapid expansion of all these branches of matrix biology is the combined result of the availability of advanced methods of cell and molecular biology and the increasing awareness of the importance of this field for a number of basic and applied sciences. This introduction is a review for the special issue of Pathologie Biologie devoted to 'Matrix Biology' and brushes an impressionistic landscape of the major advances accomplished over the finishing century and tries to predict some of the most important advances to be expected during the coming century. PMID- 11428163 TI - Structure and function in extracellular matrices depend on interactions between anionic glycosaminoglycans. AB - Connective tissue extracellular matrices define and maintain shape. Collagen fibrils transmit and resist pulling forces and soluble interfibrillar carbohydrate-rich polymers (anionic glycosaminoglycans AGAGs) resist compressive forces. This article shows that AGAGs (chondroitin, dermochondan and keratan sulphates) form supramolecular organisations which take tensile stresses as well, helping to maintain shape as part of 'shape modules'. Specific interactions with collagen fibrils tie down the AGAGs but conversely they orientate and maintain the organisation of collagen fibrils by forming interfibrillar bridges. The shapes of the AGAG chains in solution, shown by NMR to be two-fold helices, are completely complementary, allowing duplexes and higher aggregates to form spontaneously, providing participating AGAGs are oriented head-to-head i.e. antiparallel to each other. These tertiary structures are exact analogues of the beta-sheets are the basic structure of the interfibrillar bridges in the shape modules. In the absence of key shape module components, in a genetic disorder, extracellular matrices do not form an ordered tissue. PMID- 11428164 TI - Fibronectin in the vitreous body--distribution and possible functional role. AB - The presence of fibronectin in the bovine vitreous was demonstrated by immunohistochemical procedures which showed a uniform coating of the vitreous collagen network. A fractional extraction of bovine vitreous was carried out in order to determine the distribution of fibronectin and glycosaminoglycans as related to collagen fibers. About half of total fibronectin could be extracted with aqueous buffers with increasing concentrations of KCl, part of fibronectin remained however strongly associated with the insoluble collagen network even after a final extraction with 4 M urea and 0.05 M DTT. Total extractable fibronectin was of the order of 76 micrograms per vitreous, corresponding to approximately 0.17 nM fibronectin. Total quantity of GAG-s determined as uronic acid were of the order of 2200 micrograms/vitreous corresponding approximately to 4400 micrograms disaccharide units that is to about 11 nM disaccharide units of GAG per vitreous. The persistence of fibronectin, strongly associated with the collagen fibers even after repeated KCl and urea-DTT extractions was confirmed using immuno-gold labelling of vitreous collagen fibers. Gold particle density on the collagen fibers increased with the molarity of KCl used for the extractions. These findings suggest that KCl mainly removed fiber associated components probably GAG-s, which hindered the immune recognition of fiber-bound fibronectin. The strong association of fibronectin with vitreous collagen suggested a modified model for vitreous structure taking in account the binding of fibronectin both by collagen and GAG-s. PMID- 11428165 TI - Effect of procyanidolic oligomers on the permeability of the blood-brain barrier. AB - Blood-brain barrier (BBB) is the site of regulatory mechanisms which control the exchange of substances between the brain and the blood through the wall of 'true' brain capillaries with tight junctions between endothelial cells. In some pathological situations the permeability of the BBB is increased because of a partial proteolytic degradation of some constituents of the capillary basement lamina. In such cases it is important to restore normal permeability. The effect of procyanidolic oligomers (PCO) on the BBB was investigated in vivo with quantitative morphologic procedures. We also investigated the action of this drug on collagen and basement lamina constituents (Matrigel) in vitro. Collagenase injected in lateral brain ventricles was shown to increase BBB permeability. Per os administration of PCO to rats greatly increased the resistance of brain capillaries to bacterial collagenase, as shown by the inhibition of the diffusion of fluorescein-isothiocyanate-marked dextran particles from the blood-stream into the brain tissues. Calf skin collagen pretreated in vitro with PCO became more resistant to the hydrolytic action of collagenase. Similar, even more intense protective effect was seen when basal lamina constituents containing type IV collagen was incubated with PCO before exposure to pronase. These in vitro effects may partly explain the in vivo protective effect of PCO against the alteration of brain capillaries by i.v. injected bacterial collagenase. PMID- 11428166 TI - Effect of an alpha-blocker (Nicergoline) and of a beta-blocker (Acebutolol) on the in vitro biosynthesis of vascular extracellular matrix. AB - The effect of an alpha-blocking agent and of a beta-blocking agent on the biosynthesis of extracellular matrix macromolecules of the arterial wall was investigated. Rabbit aorta explants were cultured up to 48 hours with radioactive proline, lysine or glucosamine. In presence of these drugs, at concentration shown to be effective for the inhibition of platelet-endothelial cell interactions (10(-7) M), the incorporation of 14C proline in total macromolecular proline was higher than in macromolecular hydroxyproline suggesting a relatively higher rate of biosynthesis of non-collagenous proteins as compared to collagens. The alpha-blocking increased the incorporation of 14C proline in collagenous and non-collagenous proteins after 18 hours of incubation. beta-blocking also increased the incorporation of proline in macromolecular proline and hydroxyproline as compared to control cultures. Both increased the incorporation of 3H glucosamine in newly synthesised glycosaminoglycans. beta-blocking increased mainly the neosynthesis of heparan sulphate, alpha-blocking that of hyaluronan. The incorporation of 14C-lysine in crosslinked, insoluble elastin was not modified. These experiments confirm that alpha and beta-blocking agents can influence not only the tonus of aortic smooth muscle cells but also the relative rates of biosynthesis of extracellular matrix macromolecules. This effect should be taken in consideration for the evaluation of the long range effect of alpha and beta-blocking drugs on the vascular wall. PMID- 11428167 TI - Function-structure relationship of elastic arteries in evolution: from microfibrils to elastin and elastic fibres. AB - Evolution of species has led to the appearance of circulatory systems including blood vessels and one or more pulsatile pumps, typically resulting in a low pressurised open circulation in most invertebrates and a high-pressurised closed circulation in vertebrates. In both open and closed circulations, the large elastic arteries proximal to the heart damp out the pulsatile flow and blood pressure delivered by the heart, in order to limit distal shear stress and to allow regular irrigation of downstream organs. To achieve this goal, networks of resilient and stiff proteins adapted to each situation--i.e. low or high blood pressure--have been developed in the arterial wall to provide it with non-linear elasticity. In the low-pressurised circulation of some invertebrates, the mechanical properties of arteries can almost be entirely microfibril-based, whereas, in high-pressurised circulations, they are due to an interplay between a highly resilient protein, an elastomer in the octopus and elastin in most vertebrates, and the rather stiff protein collagen. In vertebrate development, elastin is incorporated in elastic fibres, on a earlier deposited scaffold of microfibrils. The elastic fibres are then arranged in functional concentric elastic lamellae and, with the smooth muscle cells, lamellar units. The microfibrils may also play a direct functional role in all mature arteries of high- and low-pressurised circulations. Finally, since blood pressure regularly increases with developmental stages, it appears possible that the early deposition of microfibrils, which are highly-conserved in evolution, corresponds, at least in part, to an early microfibril-driven elasticity in low-pressurised arteries, present across species. In vertebrates, when pressure developmentally rises above a threshold value, the vascular wall stress may turn on the expression of other resilient protein genes, including the elastin gene. Elastin would then be deposited on microfibrils and resulting in the elastic fibre network and elastic lamellae whose mechanical properties are adapted to allow for proper arterial work at higher pressures. PMID- 11428168 TI - Extracellular matrix remodeling in the vascular wall. AB - The extracellular matrix provides a structural framework essential for the functional properties of vessel walls. The three dimensional organization of the extracellular matrix molecules--elastin, collagens, proteoglycans and structural glycoproteins--synthesized during fetal development--is optimal for these functions. Early in life, the vessel wall is subjected to injury: lipid deposition, hypoxia, enzyme secretion and reactive oxygen species production during inflammatory processes, and the extracellular matrix molecules are hydrolyzed by proteases--matrix metalloproteinases, leukocyte elastase, etc. In uninjured arteries and veins, some proteases are constitutively expressed, but through the control of their activation and/or their inhibition by inhibitors, these proteases have a very low activity. During the occurrence of vascular pathologies--atherosclerosis, hypertension, varicosis, restenosis, etc.--the balance between proteases and their inhibitors is temporally destroyed through the induction of matrix metalloproteinase gene expression or the secretion of enzymes by inflammatory cells. Smooth muscle cells, the most numerous cells in vascular walls, have a high ability to respond to injury through their ability to synthesize extracellular matrix molecules and protease inhibitors. However, the three dimensional organization of the newly synthesized extracellular matrix is never functionally optimal. In some other pathologies--aneurysm--the injury overcomes the responsive capacity of smooth muscle cells and the quantity of extracellular matrix decreases. In conclusion, care should be taken to maintain the vascular extracellular matrix reserve and any therapeutic manipulation of the protease/inhibitor balance must be perfectly controlled, because an accumulation of abnormal extracellular matrix may have unforeseen adverse effects. PMID- 11428169 TI - Elastin-laminin receptor and abdominal aortic aneurysms. New subject to study? A review. AB - Abdominal aortic aneurysms and their management remain a significant health problem that is likely to assume greater importance with the expansion of the elderly population. Elastin fibres degradation and extracellular matrix remodelling seems to be the basic process in aneurysm formation. Recent investigations revealed the principal role of elastin-laminin receptor in extracellular matrix remodelling in aging and atherosclerosis. The correlation between events observed in animal aneurysm models, human aneurysms and in experiments on elastin-laminin receptor properties was discussed to propose the hypothesis about the role of elastin peptides and elastin-laminin receptor in aortic aneurysm formation. PMID- 11428170 TI - Age-related alterations in the signal transduction pathways of the elastin laminin receptor. AB - With aging we assist to alterations in the vascular structure and function. One important factor in these vascular wall changes is the degradation of the elastin fibre major protein: elastin. Elastin peptides derived from the degradation are present in human sera. Elastin peptides induce on fibroblasts, phagocytic cells, lymphocytes, smooth muscle cells and endothelial cells, a variety of biological effects mediated by the elastin-laminin receptor which has been demonstrated to be present on the membrane of these cells. The transduction pathway of the ELR receptor involves the activation of phospholipase C (PLC) by a pertussis toxin sensitive G-protein. PLC induces the production of inositol trisphosphate (IP3) leading to the increase of the intracellular free calcium on one hand, and of diacylglycerol (DAG) which stimulates the translocation to the membrane of PKC leading to the phosphorylation of members of the MAPK family, such as p42/p44 MAPK. A progressive age dependent uncoupling of the elastin-laminin receptor occurs impairing its transduction pathway and which results in alteration of the calcium signaling and loss in calcium homeostasis of the cells. These alterations in the signal transduction of the elastin-laminin receptor result in modified activities of parenchymal and phagocytic cells with aging, such as free radical production and elastase release. Thus, these age-related alterations in the elastin-laminin receptor signal transduction may be involved in the atherogenesis. PMID- 11428171 TI - Cell-matrix interactions, alteration with aging and age associated diseases. A review. AB - Cell-matrix interactions are mediated by matrix components and cell surface receptors, among them integrins. During aging there are qualitative and quantitative alterations of matrix components. Similar modifications were observed with integrins. During diseases, some integrins appear to be differentially or ectopically expressed. Two examples are given showing the appearance of key receptors and the role of antagonists in their expression opening the field of therapeutic possibilities. PMID- 11428172 TI - Corneal collagens. AB - Cornea is a highly differentiated tissue rich in extracellular matrix (ECM) specifically distributed in space in order to insure its dual role--transparency and protection of inner eye-tissues. Corneal ECM is especially rich in collagens. Since the characterisation of a number of distinct collagen types it appeared that most of them are present in the cornea. Their synthesis follows a specific program of sequential expression of the different collagen types to be synthesised during the development and maturation of the cornea. The precise regulation of the diameter and orientation of fibers, and of the interfibrillar spaces is partially at least attributed to interactions between glycosaminoglycans and collagens. The 'program' of vectorial collagen synthesis and GAG-collagen interactions changes also with age and in several pathological conditions as corneal dystrophies and wound healing. The Maillard reaction, especially in diabetes, is one of these important factors involved in age dependent modifications of corneal structure and function. Far from being inert, corneal collagens were shown to have relatively short half-lives. The biosynthesis of corneal collagens was studied also during wound healing. The refibrillation of wounded corneas does not follow the original 'program' of ECM synthesis as shown by the comparative study of wound healing using biochemical and morphometric methods. This review recapitulates briefly previous and recent studies on corneal collagens in order to present to clinicians and scientists an overview of the state of the art of this important field at the intersection of eye research and matrix biology. PMID- 11428173 TI - [Structure and regulation of articular cartilage proteoglycan expression]. AB - Beyond aggrecan, the major proteoglycan present in articular cartilage that confers resistance to compressive load and viscoelasticity to the tissue, other proteoglycan families have been described in cartilage. Among them, decorin, biglycan and fibromodulin which belong to the small leucine-rich proteoglycans family bind to matrix components, specially to collagen fibrils and thus regulate fibrillogenesis in cartilage and matrix integrity. These small proteoglycans can also interact with TGF-beta and modulate its bioavailability and stability. The third family is composed by cell surface proteoglycans as syndecans, glypican-1 and betaglycan. These molecules interact with various components of cell environment (growth factors, proteases, matrix components, etc.) and mediate numerous cell functions. Some modifications of one of these proteoglycan expression occur during degenerative pathologies and may lead to alteration of the functional properties of the tissue as well as variations in growth factor bioavailability. These factors are involved in the attempt of cartilage repair initiated by chondrocytes in the early stages of osteoarthritis. PMID- 11428174 TI - A role for estrogen in the primary prevention of Alzheimer's disease. PMID- 11428175 TI - Is low-dose hormone replacement therapy for postmenopausal women efficacious and desirable? PMID- 11428176 TI - Tibolone and its effects on bone: a review. AB - This review examines the evidence for the effects of tibolone on bone. Tibolone is a synthetic steroid with a mixed (estrogenic-progestogenic-androgenic) hormonal profile. Data suggest a complex receptor-mediated as well as metabolic regulation of the activity of tibolone at target tissue level. It has been shown that tibolone can prevent axial and appendicular bone loss induced by ovariectomy and/or a low calcium diet in young and mature rats. In addition, tibolone increases trabecular and cortical bone mineral density in rats with established osteopenia. In the rat, treatment with tibolone results in an increased strength of the femoral neck and of the vertebral body, similar to that found with estrogens. The protective effect on bone can be blocked by antiestrogens, indicating that the effect is estrogen receptor-mediated. Clinical trials have shown that loss of bone in the spine and proximal hip can be prevented with tibolone 2.5 mg/day in early- and late-postmenopausal women. In addition, a dose of 1.25 mg/day seems also to be effective, especially in late-postmenopausal women. In women with established osteoporosis, bone density of the axial and appendicular skeleton increases with tibolone. In comparative studies, tibolone 2.5 mg/day seems to be as effective as conventional hormone replacement therapy regimens. There are no direct comparative studies between tibolone and bisphosphonates or raloxifene. Furthermore, to establish the efficacy of tibolone for prevention of osteoporotic fractures, studies of the magnitude of reduction in fracture risk remain to be conducted. Finally, tibolone seems to be effective in preserving bone density in patients treated with gonadotropin-releasing hormone agonist. PMID- 11428177 TI - Relaxin: a possible future preventive therapy for cardiovascular disease in postmenopausal women and men? PMID- 11428178 TI - Effects of wild yam extract on menopausal symptoms, lipids and sex hormones in healthy menopausal women. AB - OBJECTIVES: Many women seek alternatives to hormonal therapies for the management of menopausal symptoms. Among the treatments currently popular are extracts of wild yam (Dioscorea villosa), which are applied topically in the form of a cream. These preparations are known to contain steroidal saponins, including diosgenin, which has been claimed to influence endogenous steroidogenesis. However, there have been no studies of the safety or efficacy of these preparations in the management of menopausal symptoms. METHODS: We therefore conducted a double blind, placebo-controlled, cross-over study of the effects of a wild yam cream in 23 healthy women suffering from troublesome symptoms of the menopause. After a 4 week baseline period, each woman was given active cream and matching placebo for 3 months in random order. Diaries were completed over the baseline period and for 1 week each month thereafter, and blood and saliva samples were collected at baseline and at 3 and 6 months, for measurement of lipids and hormones. RESULTS: The average age of the subjects was 53.3 +/- 1.1 (SEM) years and average time since last period 4.3 +/- 0.9 years. At baseline, the average body mass index was 27.3 +/- 0.8, cholesterol level 5.7 +/- 0.2 mmol/l and follicle stimulating hormone (FSH) level 74.2 +/- 5.1 IU/l; estradiol levels were undetectable in the majority of cases. After 3 months of treatment, no significant side-effects were reported with either active treatment or placebo, and there were no changes in weight, systolic or diastolic blood pressure, or levels of total serum cholesterol, triglyceride, high-density lipoprotein (HDL) cholesterol, FSH, glucose, estradiol, or serum or salivary progesterone. Symptom scores showed a minor effect of both placebo and active treatment on diurnal flushing number and severity and total non-flushing symptom scores, and on nocturnal sweating after placebo, but no statistical difference between placebo and active creams. CONCLUSIONS: This study suggests that short-term treatment with topical wild yam extract in women suffering from menopausal symptoms is free of side-effects, but appears to have little effect on menopausal symptoms. It emphasizes the importance of careful study of treatments for menopausal symptoms if women are to be adequately informed about the choices available to them. PMID- 11428179 TI - Dietary phytoestrogens and estrogen inhibit experimental atherosclerosis. AB - OBJECTIVE: The effect of phytoestrogen intake in combination with estrogen replacement therapy (ERT) on atherogenesis is largely unknown. The aim was thus to study the impact of phytoestrogens alone, or combined with oral estrogen, on experimental atherosclerosis in cholesterol-fed rabbits. METHODS: Two separate studies were performed in ovariectomized, cholesterol-fed female rabbits. In Study A, 45 rabbits were randomized to either a soy-free diet with or without oral 17 beta-estradiol (E2) 4 mg/day, or a soy-rich diet without any hormone for 14 weeks. In Study B, 100 rabbits were randomized into five groups (oral E2 0.5, 1, 2 or 4 mg/day, or no hormone) based on a soy-rich diet for 30 weeks. RESULTS: By the end of treatment in Study A, aortic cholesterol content was twice the amount in the group treated with the soy-free diet compared with the soy-rich group and with the soy-free plus E2 group (p < 0.001). In Study B, aortic cholesterol content showed no significant difference between the groups (ANOVA, p = 0.49), but a tendency towards a lower aortic cholesterol content in the E2 treated animals compared with placebo was observed. CONCLUSION: Dietary phytoestrogens significantly reduce aortic cholesterol content with a potency comparable to that of ERT, and seem to enhance (although mildly) the antiatherogenic effect of E2 in this model. PMID- 11428180 TI - Influence of hormone replacement therapy on postmenopausal pelvic organs. AB - OBJECTIVE: To assess the influence of hormone replacement therapy (HRT) on the pelvic organs of postmenopausal women by vaginal ultrasonography. DESIGN: The study (case-control) included 753 consecutive, postmenopausal women who were referred for routine transvaginal ultrasound examination. A total of 290 women who were using HRT at the time of examination (study group) were compared with 463 who were not treated (control group). RESULTS: The women using HRT were younger and had been menopausal for a shorter period, compared with those who had not been treated. Mean uterine volume, endometrial thickness and ovarian area were all increased in the HRT group, compared to the control group. A negative correlation was found between menopausal age and uterine volume, and ovarian area in both treated and untreated groups. However, endometrial thickness was negatively correlated with menopausal age in the untreated group only. After controlling for various parameters, a multivariate logistic analysis demonstrated that endometrial thickness was treatment status-dependent only. Uterine volume was also treatment status-dependent, but was also negatively correlated with menopausal age. As expected, the endometrium was thicker and the uterine volume was larger in the treated group. Ovarian area was not found to be treatment dependent for all menopausal ages. CONCLUSIONS: Postmenopausal women treated with HRT have a larger uterus and thicker endometrium than those of non-treated matched controls. There is a negative correlation between menopausal age and uterine and ovarian size. Endometrial thickness was found to be negatively correlated with menopausal age in only the untreated group, and is almost entirely HRT-dependent. In contrast to the uterus and the endometrium, the ovaries are not influenced by HRT. PMID- 11428181 TI - Index medicus and MEDLINE indexing for Climacteric. PMID- 11428182 TI - Clinical implications of perimenopausal steroid changes. PMID- 11428183 TI - Hormone replacement therapy and irregular bleeding. PMID- 11428184 TI - Molecular genomic characterization of the viruses of malignant catarrhal fever. PMID- 11428185 TI - Onset of accumulation of PrPres in murine ME7 scrapie in relation to pathological and PrP immunohistochemical changes. AB - In a murine scrapie model, three different methods (immunohistochemistry, Western blotting and histoblotting) for determining disease-specific PrP accumulation were compared. The incubation period of ME7 scrapie in the F1 cross of C57 BL and VM/Dk mice is about 230 days. Mice show hippocampal neuronal loss from 160-180 days post-inoculation (dpi), CA1 neuron dendritic spine atrophy at 126 dpi, and axon terminal degeneration and synaptic loss from 84-98 dpi. Infectivity titres of at least 100 are present from 40 dpi. PrP was detected immunohistochemically at 60 dpi in the hippocampus and in the thalamus. Thus, PrP accumulation in the hippocampus precedes even the earliest neurodegenerative changes. Low amounts of PrP immunolabelling were found between 60 dpi and 126 dpi, after which the intensity increased markedly. The histoblot method detected PrPres in one of four mice at 100 dpi. Western blotting of whole brains first identified the PrPres at 80 dpi. Thus, in our hands, the most sensitive method for detecting disease specific accumulations of PrP was immunohistochemical examination. However, immunohistochemical methods are unable to distinguish the normal and abnormal isoforms of PrP. It is therefore possible that the initial accumulation of PrP takes place as PrPsen and that the translation of PrPsen to PrPres does not take place until the later stages of the disease process. The accumulation of disease specific PrP lags behind the development of infectivity titres. The relative rates of increase of infectivity titre and PrP accumulation are different, suggesting that these parameters may be measures of different biological events. PMID- 11428186 TI - Atypical cilia in the bronchiolar epithelium of pigs experimentally infected with hog cholera virus. AB - To study the effect of hog cholera virus on the epithelial cells of the bronchiolar mucosa, 12 pigs were inoculated with a highly virulent strain. Immunohistochemical and ultrastructural examination of the ciliated epithelial cells demonstrated an increase in the number of atypical cilia. The latter showed alterations in the microtubular pattern, possibly resulting from viral interference with the normal metabolism of the epithelial cells. PMID- 11428187 TI - Immunohistochemical study of age-dependent brain lesions in mice infected intracerebrally with Kasba (Chuzan) virus. AB - When Kasba (Chuzan) virus (an orbivirus) was injected intracerebrally into 1-, 2- or 4-week-old mice, non-purulent necrotizing encephalitis developed and the mice showed nervous symptoms and became moribund. The necrotic lesions were more severe in younger animals. In 1-week-old mice, viral titres rose until 7 days post-infection, while in 2- and 4-week-old animals the titres reached a peak on day 3 and then declined gradually. Glial fibrillary acidic protein-positive astrocytes increased in the white matter, hippocampus and subpial area of the cerebral cortex of infected animals, and lectin-RCA-1-positive cells, thought to be microglial cells, increased in the necrotic lesions. The number of these glial cells increased even after viral titres had declined. In this study there were no survivors in any age group, but survival time increased with age. PMID- 11428188 TI - Lymphocyte subtypes in experimentally induced early-stage bovine tuberculous lesions. AB - The identity of the lymphocyte subtypes constituting the lymphocytic mantle within developing early-stage lesions of bovine tuberculosis was investigated immunohistochemically in calves inoculated intranasally with 2 x 10(7) colony forming units of a field isolate of Mycobacterium bovis. Pulmonary lesions were examined 7, 14, 21, 28 and 42 days after inoculation, and bronchial lymph node lesions at 35 days. The immunolabelling results reported were obtained with monoclonal antibodies against two T-cell epitopes (WC1+ gamma delta and CD2+) and against B-cell epitopes. Large numbers of CD2+ T-lymphocytes were observed around developing areas of necrosis throughout the study; WC1+ gamma delta cells, however, were more numerous at these sites up to and including day 21. On the other hand, aggregates of B lymphocytes did not become prominent in areas adjacent to lesions until day 42. The results suggest that these lymphocyte phenotypes play a role in the pathogenesis of early-stage lesions. PMID- 11428189 TI - Histological observations on the brains of symptomless 7-year-old cattle. AB - The histological changes in the brains of 506 clinically normal 7-year-old cattle, which were part of a cohort study on maternal transmission of bovine spongiform encephalopathy, are described. Vacuolation of the white matter, of unknown aetiology, located particularly in the substantia nigra, was a frequent finding. Vacuolated neurons were commonly observed in the red nucleus (64.3% of the animals) and in the habenular nucleus (50.1%). Spheroids were found in 10.8% of the brains, most frequently in the vestibular nuclei. Cellular inflammatory infiltrates in association with blood vessels occurred in 30% of the animals at various locations in the brain; their aetiology remains uncertain, but they may have reflected subclinical or latent infections. Mineralization of the wall of blood vessels, with proliferation of the intima, was observed frequently in vessels of the internal capsule and was probably associated with ageing. The description of histological findings in the brain of symptomless adult cattle in the present study provides a useful background for diagnostic bovine neuropathology. PMID- 11428190 TI - Immunohistochemical analysis of macrophages and myofibroblasts appearing in hepatic and renal fibrosis of dogs. AB - Some peptide growth factors produced by macrophages play a role in fibrosis following tissue injury, through the induction of myofibroblasts. In the present study, the appearance of macrophages and myofibroblast development in hepatic and renal fibrosis was determined by immunohistochemical analysis of tissue from 15 dogs. The hepatic and renal fibrosis was classified as grade I, II or III, depending on the extent (percentage) of fibrotic areas per unit area measured by morphometry with Azan-stained sections. The presence of alpha-smooth muscle actin immunolabelled myofibroblasts was directly related to advancing grade of both hepatic and renal fibrosis. Lysozyme-immunolabelled macrophages also increased in number with increasing grade of hepatic and renal fibrosis. These findings indicate that myofibroblasts and lysozyme-positive macrophages may contribute to progressive fibrosis in canine liver and kidney disease. Interestingly, the number of macrophages recognized by AM-3K, a newly generated monoclonal antibody capable of labelling exuded macrophages and resident tissue macrophages in dogs, fell significantly in grades II and III of renal fibrosis. By contrast, in hepatic fibrosis there were no marked differences in the number of AM-3K-positive macrophages between grades. These findings suggest that there are functional differences between lysozyme- and AM-3K-positive macrophages. PMID- 11428191 TI - Expression of cytokeratins and vimentin in normal and neoplastic tissue from the bovine female reproductive tract. AB - The distribution of cytokeratins (CKs) and vimentin in the normal genital tract of calves and cows at different stages of the oestrous cycle and in epithelial tumours of the tract was studied immunohistochemically. Few differences in CK and vimentin immunolabelling were detected in relation to age or stage of the oestrous cycle. Coexpression of CKs in simple epithelia and in basal cells of stratified epithelia was detected in the oviduct and endocervix; this coexpression was different from that previously described in women. The demonstration of CKs but not vimentin in the neoplastic cells of a serous superficial ovarian papilloma suggested an origin from the ovarian surface epithelium, while the coexpression of CKs and vimentin in serous papillary and mucinous cystadenomas pointed to a possible origin from the rete ovarii. Studies on three uterine adenocarcinomas and the ovarian metastases from two of these showed an endometrial-CK phenotype. The intermediate filament profile of normal endometrium, conserved in uterine adenocarcinomas and their ovarian metastases, may be useful in discriminating between ovarian metastases from endometrial carcinomas and those originating from primary carcinomas in other organs. PMID- 11428192 TI - Sertoli cell tumour in an Amur tiger. AB - The histological and immunohistochemical characteristics of a malignant Sertoli cell tumour in a 17-year-old Amur tiger (Panthera tigris altaica) are described. Histological examination of the primary lesion in the right testis and metastatic lesions throughout the internal organs showed a variable cellular pattern with an admixture of tubular structures divided by fine stroma filled with fusiform to stellate cells, and sheets of polygonal cells with abundant vacuolated cytoplasm. Immunohistochemical techniques demonstrated strong positive staining for neuron specific enolase and variable positive staining for vimentin in neoplastic cells, supporting a diagnosis of a tumour of Sertoli cell origin. PMID- 11428193 TI - The anti-human CD21 antibody, BU33, identifies equine B cells. AB - The number of antibodies for identifying equine B cells is small and the number that react with well-defined epitopes even smaller. The monoclonal antibody, BU33, which is directed against human CD21 (Complement Receptor 2; CR2) was shown to identify (1) follicular lymphocytes in the lymph nodes and spleen of three horses, and (2) a mean of 18 +/- 6% (SEM) of peripheral blood lymphocytes from 10 horses. These findings indicate that the antibody identifies equine B cells and possibly equine CR2 or a related molecule. This study adds to the reagents available for equine research and diagnostic pathology. PMID- 11428194 TI - [Children's health: trends and determinants in the city of Sao Paulo, Brazil, in the second half of the 20th Century]. PMID- 11428195 TI - [Secular trends of height in adulthood of children born in the city of Sao Paulo, Brazil from 1950-1976]. AB - OBJECTIVE: The purpose of this study was to estimate the secular trend/change in the height of young males born in the city of S. Paulo between 1950 and 1976 and measured in the year they turn 18 years. METHODS: A random and representative sample (6,942 individuals) was evaluated in military draft. Statistical analyses included Shapiro-Wilk test for normality of height distribution in each birth cohort, and linear regression analysis for trend on heights. RESULTS/CONCLUSIONS: Stature has increased 3.42 centimeters during the 27 years covered by the study (1.26 cm/decade). The trend was not linear: in the 50s, there was a statistically significant increase (0.84 cm/decade); in the 60s, a smaller but non-significant increase (0.5 cm/decade) was seen; in the period of 1970-76, a greater increase in heights (2.9 cm/decade) was observed. The secular change rate observed was comparable to the rate seen in other countries. The most recent birth cohorts (1975 and 1976) achieved the higher statures in the study (approximately 175 cm). Despite these height increments, deficits of 1.8 e 6.2 cm were seen when the taller cohorts of the study were compared to American young males born in 1961 (NCHS) and Dutch men born in 1972. If there won't be any changes in the accelerated rates of the 70s, young people of Sao Paulo may overcome these deficits in about one or three decades. PMID- 11428196 TI - [Evolution of maternal and child health care in Sao Paulo (1984-1996)]. AB - OBJECTIVE: Two consecutive household surveys undertaken in mid-80s and mid-90s in the city of S. Paulo, Brazil, made possible to establish time trends of several child health determinants and indicators as well as to analyse the relationships among them. The study intends to report trends in maternal and child health care. METHODS: Random samples of the population aged from zero to 59 months were studied: 1,016 children in the period of 1984-85 and 1,280 children in 1995-96. Both surveys investigated three components of maternal and child health care: prenatal care, delivery and newborn care and routine health care provided to children up to five years of age (including development follow-up and vaccination). RESULTS: Favourable changes seen in the period between the two surveys were the continuing universal birth coverage, significant increase in rooming-in in maternity hospitals and the number of routine visits for babies in their first year of life and, particularly, the universal outreach of the DPT, measles and tuberculosis vaccinations. Unfavourable trends were seen regarding the slight and clearly insufficient growth of prenatal care, the still high percentage (of near 50%) of cesarean sections, and the limited routine visits for children after their first year of age. CONCLUSIONS: Estimates in the same period for the outreach of maternal and child health care in other urban areas of Brazil reinforce the unsatisfactory trends of the prenatal care in S. Paulo. Favourable comparisons are only seen regarding the outreach of child vaccination. The influence that changes in the maternal and child health care provided in the city may have exerted on several child health indicators is examined in subsequent articles. PMID- 11428197 TI - [Secular trends in birth weight in the city of Sao Paulo, Brazil (1976-1998)]. AB - OBJECTIVE: Data from two household surveys on infant and child health status undertaken in the mid-80s and mid-90s, complemented with previous data collected from maternity hospitals records and more recent data provided by the state system on birth registries, allowed to characterize and analyse secular trends in birth weight in the city of S. Paulo, Brazil. METHODS: The household surveys included random samples of children under 5 years old (n = 1,016 children in 1984 85 and n = 1,280 children in 1995-96). A random sample of births that took place in the city's hospitals in the year of 1976 (n = 5,734) was drawn from the hospital records. Birth registries refer to children born in the city between 1993 and 1998 (around 200,000 per year). The study of the social distribution of birth weight took into account the per capita family income and maternal schooling. For the analysis of the determinants of secular trends, hierarchical causal models, multivariate regression analyses and calculations analogous to the ones used to assess population attributable risks were applied. RESULTS/CONCLUSIONS: Birth weight distribution in S. Paulo city (an average of 3,160 g and 8.9% of the values < 2,500 g) is below the expected for optimum fetal growth conditions (average 3,400-3,500 g and 4-5% of the values < 2,500 g). The birth weight distribution did not change substantially along the study period (1976-1998). However, there are evidences of changes when different socioeconomic strata are considered separately. Among the lower strata trends have been positive and this seems to be due to increases in intrauterine growth as a result of an improvement in family's purchasing power, women's weight and height, prenatal care and, possibly, the reduction in smoking. Among the higher socioeconomic strata, birth weight trends have been negative apparently due to an increase in premature births of unknown origin. PMID- 11428198 TI - [Secular trends in postanal growth in the city of Sao Paulo, Brazil (1974-1996)]. AB - OBJECTIVE: Data from three household surveys undertaken in the city of S. Paulo, Brazil, from mid-80s to mid-90s allow to characterize and analyse secular trends in infant and child linear growth. METHODS: In the three surveys, random population samples aged from zero to 59 months (1,008 children in the period of 1974-75; 1,016 in 1984-85 and 1,280 in 1995-96) were studied. Recumbent length was obtained from children up to 24 months of age and then stature was recorded. The international growth standard was used to evaluate the child's height according to age and sex. For the study of the social distribution of growth status, tertiles of the per capita family income was taken into account in each survey. For the study of the determinants of secular trends, hierarchical causal models, multivariate regression analyses and calculations analogous to the ones used to assess population attributable risks were applied. RESULTS/CONCLUSIONS: In the period of 22 years covered by the three surveys, the secular trend of child growth in S. Paulo City showed a positive, continuous and apparently uniform curve. It was equivalent to a total average gain of 0.650 z scores of the international growth reference, or near 2.3 cm at the age of 30 months. The higher gain was observed among the third poorest fraction of the population--3.3 cm--and the lower gain--1.7 cm--among the third richest. Positive changes in distal determinants (family income and maternal schooling) and intermediate determinants (housing, sanitation, access to health services and reproductive past history) of child growth explained substantially part of the improvements seen from the mid-80s to mid-90s. PMID- 11428199 TI - [The study of secular trends of health indicators as a strategy in epidemiologic investigation]. PMID- 11428200 TI - [Secular trends in malnutrition and obesity among children in the city of Sao Paulo, Brazil (1974-1996)]. AB - OBJECTIVE: Data from three household surveys undertaken in the city of S. Paulo, Brazil, from mid-80s to mid-90s allow to characterize and analyse secular trends in malnutrition and obesity among infants and children. METHODS: The three surveys included random population samples aged from zero to 59 months (1,008 children in the period of 1974-75; 1,016 in 1984-85 and 1,280 in 1995-96). The malnourished status was diagnosed when height-for-age and/or weight-for-height indices were below two standard deviation of the mean values expected according to the international growth reference. The obesity status was determined when weight-for-height indices fell two standard deviations above the reference. The study of the social distribution of malnutrition and obesity in each survey took into account tertiles of the per capita family income. For the study of the determinants of secular trends, hierarchical causal models, multivariate regression analyses and calculations analogous to the ones used to assess population attributable risks were applied. RESULTS/CONCLUSIONS: In the period of 22 years covered by the three surveys, child malnutrition was controlled in the city and became relatively rare even among the poorest families. The risk of obesity remained low and restricted to the richest families. Positive changes in distal (family income and maternal schooling) and intermediate determinants (sanitation, access to heath services and reproductive past history) of child nutritional status substantially explained part of the decline in the prevalence of malnutrition seen in the mid-80s to mid-90s. PMID- 11428201 TI - [Secular trends in childhood in the city of Sao Paulo, Brazil (1984-1996)]. AB - OBJECTIVE: Data from two consecutive households surveys undertaken in mid-80s and mid-90s allow to characterize and analyse secular trends in infant and child anaemia in the city of S. Paulo, Brazil. METHODS: The two surveys included random population samples aged from zero to 59 months (1,016 in the period of 1984-85 and 1,280 in 1995-96). Capillary blood samples, collected by digital puncture in the two surveys, were analysed regarding their haemoglobin concentration. The anaemic status was determined when haemoglobin concentration was below 11 g/dL. For each survey, the study of the social distribution of child anaemia took into account tertiles of the per capita family income. For the study of the determinants of secular trends, hierarchical causal models, multivariate regression analyses and calculations analogous to the ones used to assess population attributable risks were applied. RESULTS/CONCLUSIONS: In the time span from the first to the second survey, there was a significant reduction in the average haemoglobin concentration (from 11.6 g/dl to 11.0 g/dl), as well as a considerable increase in anaemia prevalence (from 35.6% to 46.9%). Unfavourable trends were observed in both sexes, all age groups and all income strata. Trends were still less favourable among the poorest families, aggravating the social burden related to child anaemia. Changes in distal (family income and maternal schooling) and proximal determinants (breast or bottle-feeding) of child anaemia were positive in the study period and therefore they cannot explain the increase in the disease. A low iron diet could explain the high prevalence of anaemia in both surveys but could not explain its further increase. PMID- 11428202 TI - [Evolution of socioeconomic determinants of health in childhood in the city of Sao Paulo, Brazil (1984-1996)]. AB - OBJECTIVE: Two consecutive household surveys undertaken in the mid-80s and mid 90s in the city of S. Paulo, Brazil, made possible to establish time trends of several child health determinants and indicators as well as to analyse the relationships among them. The study intends to report trends in socioeconomic determinants of child health. METHODS: Random samples of the population aged from zero to 59 months were studied: 1,016 children in the period 1984-85 and 1,280 children in 1995-96. Both surveys investigated the family's per capita income and the mother's years of schooling. Nominal incomes were deflated according to the National Consumer Prices Index and expressed as values of October 1997. RESULTS: From mid-80s to mid-90s average family income doubled and the proportion of low income families was reduced by 50% while average maternal schooling increased by 1.5 years and maternal illiteracy was almost eradicated. Income concentration increased in the period. CONCLUSIONS: Increases in income and schooling are higher than those reported for the whole population in the country, which may be attributed to selective declines in fertility among the city's poorest families. The influence that changes in family income and maternal schooling may have exerted on several child health indicators is examined in subsequent articles. PMID- 11428203 TI - [Secular trends in diarrhea disease of childhood in the city of Sao Paulo, Brazil (1984-1996)]. AB - OBJECTIVE: Data from two consecutive household surveys undertaken in mid-80s and mid-90s allow to characterize and analyse secular trends in infant and child diarrhea in the city of S. Paulo, Brazil. METHODS: The two surveys included random population samples aged from zero to 59 months of age (1,016 in the period of 1984-85 and 1,280 in 1995-96). In both surveys the instant prevalence of diarrhea (proportion of examined individuals reporting three or more episodes of liquid stools in 24 hours) and the annual incidence of hospitalizations due to the disease were calculated. These two indicators were estimated from household interviews conducted by pediatricians with the children's mothers. In both surveys the interviews were distributed throughout a period of almost 12 months to assure a uniform coverage of the various areas of the city during the four seasons. For each survey, the study of the social distribution of the disease took into account tertiles of the per capita family income. For the study of the determinants of secular trends, hierarchical causal models, multivariate regression analyses and calculations analogous to the ones used to assess population attributable risks were applied. RESULTS/CONCLUSIONS: In the time span from the first to the second survey, there was substantial reduction in both the prevalence of diarrhea (from 1.70% to 0.90%) and the hospitalizations due to the disease (from 2.21 to 0.79 hospitalizations per 100 children-year). A more significant reduction was observed among the third poorest families, narrowing the social gradient relative to the disease. An increase in family income and improvement in water supply could substantially explain part of the decline in the disease and, for children under two years of age, a discrete increase in breast-feeding may have also played a positive role. PMID- 11428204 TI - [Value of imaging in acute appendicitis diagnosis in the adult: the surgeon's point of view]. PMID- 11428205 TI - [Uterine artery embolization for symptomatic uterine fibromas. Informed consent of the patient]. AB - It would seem necessary that an information sheet and written consent should be exchanged and signed by the physician and the patient, especially when interventional radiology procedures are involved. A comprehensive interview with full explanation is also mandatory if the patient is to be adequately informed. The existence of a specific document provides two benefits: For the patient: an understanding of the procedure and of the associated risks and benefits tends to reduce the natural anxiety which stems from ignorance and better prepares her to make an informed decision on whether to agree (i.e. give consent) to the procedure or decline it. For the physician: the document represents a formal way of demonstrating that he or she has fulfilled his or her duties in the matter of patient information and consent. A written form would seem to be the best solution to meet these requirements. A document of this type is proposed in this case in the form of a series of questions and answers about embolization of the uterine arteries as a modality for the treatment of symptomatic uterine fibroids. PMID- 11428206 TI - [Evaluation of diagnostic methods]. AB - AIM: Describe the statistical tools for the evaluation of a diagnostic test. MATERIAL AND METHODS: Description of the methods and practical examples based on published data. RESULTS: The following methods are described: 1) reproducibility of a measurement, both for a qualitative and a quantitative result, 2) comparison of a new diagnostic test to a reference test, 3) comparison of two diagnostic tests, 4) sample size computation. CONCLUSION: The tools required to evaluate diagnostic tests rigorously are available and simple. They should be used more often. PMID- 11428207 TI - [Normal bone marrow: dynamic aspects in magnetic resonance imaging]. AB - The bone marrow is a complex organ that contains fat and nonfat cells, the proportion of which varies greatly with age and in the different bones of the skeleton. Magnetic resonance imaging provides information on the composition of the medullary cavity of any given bone and on the distribution of red and yellow marrow in the skeleton. The wide spectrum of appearances of the normal bone marrow at MR imaging will be reviewed. The purpose of this paper is to determine the MR appearance of the bone marrow, to illustrate the phenomenon of marrow conversion and to familiarize the readers with the complex parameters that interfere with the MR appearance of normal bone marrow. PMID- 11428208 TI - [Value of local ophthalmic artery fibrinolysis in severe forms of central retinal vein occlusion]. AB - PURPOSE: To assess the value of selective ophthalmic artery thrombolysis as a treatment for central retinal vein occlusion (CRVO) for which no alternative therapy is available. MATERIALS AND METHODS: Patients included in this study presented with recent severe non-ischemic CRVO. Urokinase (300,000 IU) was perfused for 40 minutes into the ophthalmic artery. Visual acuity, fundoscopy and retinal arteriovenous transit time were assessed during one year of follow-up. RESULTS: Five of the 13 patients treated experienced a marked improvement of vision (p = 0.05) and retinal perfusion within 24-48 hours, and exhibited progressive lesion regression at fundoscopy within 2-4 weeks. The clinical course of the 5 patients prior to treatment resembled that of combined central retinal artery and vein obstruction (CRAO/CRVO), which typically has a poor visual outcome. One patient relapsed 1 month after thrombolysis. No technical complications were observed. CONCLUSION: Although there was no control group, the short period between fibrinolysis and significant visual improvement combined with marked retinal perfusion improvement, suggests that local thrombolysis is beneficial for CRVO, especially recent CRAO/CRVO. PMID- 11428209 TI - [Helical CT and diagnostic evaluation of cranio-facial fibrous dysplasia]. AB - PURPOSE: To evaluate the role of helical CT in the evaluation of skull abnormalities seen in fibrous dysplasia. MATERIALS AND METHODS: Prospective study on 6 cases with cranio-facial fibrous dysplasia (3 males and 3 females) aged 8 to 59 years old, evaluated with plain radiographs, helical CT and, in 1 case, MRI. Helical CT was performed in 3 cases on a CT Twin scan (Elscint), in 3 cases on a Somatom Plus scan (Siemens). Multiplanar reconstructions and 3D reconstructions were done in all cases. 4 cases had a histopathological confirmation of fibrous dysplasia lesions by bone-biopsy (3 cases) or surgical biopsy (1 case). RESULTS: The 6 cases included sclerotic form in 3 cases, cystic variety in 1 case, polyostotic variety in 1 case, mixed form in 1 case. Topographic localizations of fibrous dysplasia were skull base involvement in 1 case, multizonal skull involvement in 4 cases, maxillo-mandibulary involvement (cherubism) in 1 case. CONCLUSION: Helical CT is an optimal method to evaluate the skull lesions of fibrous dysplasia. The review of source images must always be done. 3-dimensional bone reconstructions are better to visualize the anterior cranial vault and skull base dysmorphy. Helical acquisition is a compromise in case by case between the dose of irradiation and the quality of source images. PMID- 11428210 TI - [Imaging of the intrahepatic biliary tree with thick slice MR cholangiography]. AB - PURPOSE: To assess the value of single shot fast spin echo MR sequence (SS-FSE) in the evaluation of the normal and pathologic intrahepatic biliary tree. MATERIAL AND METHODS: 418 consecutive patients (457 examinations) referred for clinical and/or biological suspicion of biliary obstruction underwent MR cholangiopancreatography (MRCP). All patients were imaged with a Signa 1.5 T GE MR unit, with High Gradient Field Strength and Torso Phased Array Coil. Biliary ducts were imaged with SS-FSE sequence, coronal and oblique coronal 20 mm thick slices on a 256 x 256 matrix. Total acquisition time was 1 second. Source images were reviewed by two radiologists blinded to clinical information. In case of disagreement, a third radiologist's opinion was requested. In all cases, MRCP results were compared with direct biliary tract evaluation, other imaging studies and clinical and biological follow-up. RESULTS: In all cases, MRCP produced high quality images. Numerous branch of division were observed although the peripheral intrahepatic ducts were well seen in more than 90% in an area 2 cm below the capsule. The number of division was statistically higher when mechanical obstruction was present. Intrahepatic calculi or peripheral cholangiocarcinoma were well detect by MRCP. For the detection of cholangitis, MRCP sensitivity was 87.5% but the positive predictive value was only 57.7% because of a high number of false positive. The diagnosis of primary sclerosing cholangitis must be made only on strict criteria and slightly dilated peripheral bile ducts unconnected to the central ducts in several hepatic segments were a characteristic MR sign of primary sclerosing cholangitis. CONCLUSION: MRCP can be proposed as a first intention imaging technique for the evaluation of intrahepatic ducts. PMID- 11428211 TI - [Localization of non-palpable breast lesions with black carbon powder (experience of the Catholic University of Louvain)]. AB - PURPOSE: The authors report their experience in presurgical localization of non palpable lesions with black carbon powder. MATERIALS AND METHODS: 153 cases of primary tumorectomies and mastectomies have been reviewed. The suspension is prepared in the hospital's pharmacy, the procedure is performed days or weeks before scheduled surgery without diffusion in the surrounding tissues. RESULTS: In 92% of cases, the carbon marker was observed less than 5 mm from the target and no significant interference with the pathologic diagnosis has been observed. CONCLUSION: Patient tolerance is good and providing close collaboration between the different members of the treating team, the authors believe that this technique may be a good alternative to wire localization. PMID- 11428212 TI - [Mazabraud syndrome: a case diagnosed with MRI]. AB - We report a case of Mazabraud's syndrome diagnosed with MRI in a 40-year-old woman. PMID- 11428213 TI - [Coronary steal syndrome by systemic pulmonary hypervascularization fed by a mammary-left anterior descending artery bypass. Treatment by embolization]. AB - We report a case of coronary-steal syndrome which occurred after coronary bypass surgery. It was related to systemic hypervascularization of the lung caused by a bronchopathy. The steal syndrome was fed by an ectopic bronchial artery arising from the internal mammary--left anterior descending artery bypass graft. The myocardial ischemia disappeared after hyperselective embolization of the ectopic bronchial artery. The authors outline the rarity of this syndrome and its pathophysiology. They insist on the necessity to perform broncho-systemic arteriography for candidates to coronary surgery, in patients with thoracic diseases which can induce systemic hypervascularization of the lung. PMID- 11428214 TI - [Post-traumatic rupture in ureteropelvic junction obstruction syndrome: two case reports]. AB - Post-traumatic rupture of UPJ obstruction is a rare event, with few reported cases in the literature. Diagnosis is suggested on imaging studies, especially US and CT findings. The presence of an anterior pelvic hematoma associated with thinning of kidney parenchyma, very distended pelvis and non dilated ureter is suggestive of pre-existing pathology. PMID- 11428215 TI - [Malignant bilateral mediastinal teratoma: a case report]. AB - The authors report a case of bilateral malignant teratoma of the mediastinum in a 40 year old female presenting with chest pain. Two separate bilateral soft tissue masses were noted in the mediastinum at imaging. On the right side, the mass was heterogeneous with large calcification and infiltration of the pericardium. On the left side, the mass was well defined, heterogeneous and without evidence of local invasion. The analysis of samples obtained by US guided puncture biopsy and surgical specimen of the two masses confirmed the diagnosis of pluricellular malignant teratoma. Mediastinal malignant teratomas represent 1 to 5% of all mediastinal tumors. To our knowledge, no case of synchronous bilateral mediastinal teratoma has never been reported. PMID- 11428216 TI - [MRI of an afferent loop syndrome presenting as obstructive icterus]. AB - The afferent loop syndrome corresponds to an acute or chronic obstruction of the afferent loop following a partial gastrectomy with Billroth II gastro-jejunal anastomosis. We describe the case of a 77-year-old man with history of partial gastrectomy for peptic ulcer disease performed 31 years ago and currently admitted for jaundice and poor general status. MR imaging showed dilatation of biliary and pancreatic ducts and showed a soft tissue mass between the afferent loop and the residual stomach. Endoscopy showed complete obstruction of the afferent loop by a biopsy-proven adenocarcinoma. The patient died of sepsis shortly after endoscopy of septicemia. PMID- 11428217 TI - [Quid? Intestinal invagination]. PMID- 11428218 TI - [About quid "abdominal textiloma. Associated intestinal fistula"]. PMID- 11428219 TI - Serum lipid levels and M/L55 allele distribution of HDL paraoxonase gene in Saami and Finnish men. AB - Paraoxonase (PON) is an antioxidative enzyme, which eliminates lipid peroxides. The mutation in codon 55 of PON1 gene causes a change of methionine (M-allele) to leucine (L-allele) and influences PON activity. The Saami are a population living in the northern part of Fennoscandia. In previous studies their death rate from coronary artery disease (CAD) was found to be low. We compared PON M/L55 allele frequencies of 68 Saami and 68 Finnish men and related the PON genotypes to plasma lipid levels and to the levels of autoantibodies against oxidized LDL. The M/L55 genotypes were determined by PCR and restriction enzyme digestion. ELISA was used to measure antibodies against oxidized LDL. The L- and M-allele frequencies were 64% and 36% in Saami population and 64% and 36% in Finnish men, respectively (p = NS, Fisher's exact test). There were also no significant differences in plasma lipid levels or in antibody levels against oxidized LDL between PON genotypes or between Saami and Finnish men. Our results indicate that the PON M/L55 genotype is not associated with plasma lipid levels or the levels of autoantibodies against oxidized LDL in these populations. The Saami men have the same PON M/L55 allele distribution as the Finnish men and the PON genotype might thus not be one factor protecting Saami against CAD. PMID- 11428220 TI - Modulation of atherosclerotic risk factors by seal oil: a preliminary assessment. AB - We examined whether dietary supplementation with seal oil influenced the risk factors of atherosclerosis in healthy volunteers. Two intervention studies were carried out as preliminary steps in a larger project which aim at elucidating the disease preventive potential of seal oil. In study I ten healthy volunteers added 10 capsules of seal oil to their normal Western diet for six weeks. Blood tests were analysed for total-, HDL-, and LDL-cholesterol and plasma triglyceride, and the ratio of n-6/n-3 fatty acid was determined in plasma and erythrocyte membranes. In study II we examined the effect in five healthy volunteers who had only 5 capsules of seal oil daily for six weeks. As an additional test in study II, the effect on the proinflammatory TNF-alpha cytokine in lymphocytes was determined. A slightly decreased, however, not significant effect was observed for each of the cholesterol's after seal oil supplementation. In both studies plasma triglyceride, and the n-6/n-3 fatty acid ratio of plasma and erythrocytes were significantly reduced upon seal oil intake. During the intervention period of study II a distinct reduced level of TNF-alpha was observed in isolated lymphocytes. The examinations suggest that supplementation of seal oil, 10 capsules or 5 capsules/day, may have beneficial effects on factors thought to be associated with cardiovascular and thrombotic diseases. PMID- 11428221 TI - Seasonal fluctuations in hospitalisation for pneumonia in Finland. AB - The purpose of this paper is to describe seasonal fluctuations in hospitalisation for pneumonia in Finland over the period 1972-1993. Treatment periods with pneumonia as the main or secondary diagnosis were collected from the national hospital discharge register. The seasonality of monthly hospital admissions was analysed using the X11 ARIMA procedure. The population of Finland numbered 5 million during the period concerned, and there were a total of 453,393 pneumonia treatment periods, 17.9% in the age-group under 16 years, 29.6% among working-age persons and 52.5% among persons aged over 64 years. The number was greatest in the total population in December (20.5%) and January (25.1%) and lowest in July ( 19.7%) and August (-24.3%). Seasonal fluctuations in pneumonia hospitalisations pointed to the same trend in both the sexes, in that the number increased in winter and decreased in summer. Children's admissions began to increase a month earlier than those of the other age groups, however, in November, while a transient increase in admissions of working age persons in March is attributable to the more extensive use of hospital services by army conscripts. Admissions of persons aged over 64 years fell more slowly from the winter peak than did those of the other segments of the population. The winter peak in hospital admissions for pneumonia can be anticipated administratively by increasing the number of hospital beds and intensifying outpatient care. It may not be necessary to increase health care resources to achieve this, however, as the necessary resources can be obtained through reductions in the summer months. Proper recognition of admission peaks, the points in time at which these regularly occur and the target groups within the population can contribute greatly to health care planning and its economy. PMID- 11428222 TI - The effect of season, occupation and repeated winterings on anthropologic and physiological characteristics in Russian antarctic staff. AB - Thirty anthropometric and ten physiological parameters were examined over a 10 month-period in 1985-86, in 66 male polar explorers, aged 25-61 years, at an Antarctic station (Mirny observatory). For 30 of these persons this was their first wintering in Antarctic while the remaining 36 had wintered there at least once before. The mentioned measurements were taken on 3 different occasions in April, September and January corresponding in the Antarctic to the beginning of the polar night and interim season period and the beginning of the polar day. Subjects of the investigation belonged to 3 occupational groups: administrative personnel, scientific staff, and manual laborers. Extended statistical analysis of the data was carried out in an attempt to distinguish the dynamics of the studied parameters in relation to the season of wintering (Climate), the number of previous winterings (Frequency), the type of occupation (Work), and their interactions. Multifactorial statistical analyses were also performed, so as to adjust for age of subjects, which is a requisite for evaluating the factor of repeating winterings, obviously age-related. Changes in a number of characteristics were clearly recognized as connected with the factor of repeating winterings, to wit: 1) Anthropometric parameters such as or subcutaneous fat and relative muscle mass; 2) Parameters pertaining to speed of neuromuscular response such as wrist muscle effort, or time of simple motor response; and 3) Physiological parameters encompassing vascular--blood pressure and respiratory- spirometric measurements. CONCLUSION: Our findings suggest that frequency of repeated winterings is not a leading factor influencing physical status of examinees. Occupational status of examinees alone or in interaction with repeated winterings and seasonal climatic factors have more prominent impact on the well being of polar explorers. PMID- 11428223 TI - Family as a child development context and smoking behaviour among schoolchildren in Greenland. AB - Smoking among greenlandic schoolchildren is considered a major health hazard. Greenland is a modern society susceptible to social changes, including changes in family structure. The objective was to investigate if changes in the family, as a child development context, were affecting schoolchildren's smoking behaviour. The survey was carried out in February 1998 in all schools in Greenland in the context of the WHO Health Behaviour in School-Aged Children study. 3081 questionnaires were received, which gave a response rate of 68% of the total study population. The present sample from the HBSC study consists of 1648 students, 826 boys and 822 girls in the age groups 11, 13 and 15 years old. Descriptive statistics and a logistic regression model were applied. The results were that smoking prevalence among Greenlandic schoolchildren increased with age and was lower in boys than in girls. Daily smoking prevalence among 15 year olds was high, namely 50% among boys and 56% among girls. With regard to family structure, it was found that the odds ratio for smoking was higher when the child was living in an broken/restructured family versus living in a biological family. The influence of socio-economic class on smoking behaviour was weak and statistically insignificant, and age proved to be the strongest predictor of prevalence in smoking behaviour. PMID- 11428224 TI - Physical development in Tuvinian adolescents. AB - It was found that mean values of height and mass in 13-14-year-old indigenous Tuvinian children were increased for the last 26 years. We have found gender difference in a chronological dynamics of a yearly body mass and height increases. As for the last ten years, these changes correspond to a harmonic physical development only for the boys of 13. At the same time we observed an insufficient increase in body mass and significant blood pressure increase in senior boys. A yearly percentage of the increase in blood pressure in junior boys was the same as mass increase, but among the senior children the increase in blood pressure was greater compared with mass increase. In the 70-80s, the indices of body mass and height increases for Tuvinian girls and boys were the same. PMID- 11428225 TI - The utilization of antenatal services in remote Manitoba First Nations communities. AB - The purpose of this study was to describe the utilization of antenatal services by First Nations women in four northern Manitoba communities between January, 1996 and December 1996, and to explore possible relationships between the women's behaviors and antenatal clinic attendance. This study indicated that First Nations women received an optimal level of antenatal service. On average, the women first came to the nursing station in the ninth week of pregnancy and saw the health care provider generally a nurse, ten times for routine visits before maternal evacuation. A minority of women, however, had fewer than five visits. The frequency of routine antenatal clinic attendance was explained by the linear multiple regression model. A higher number of past pregnancies was associated with a decrease in the number of routine antenatal visits, while the number of pregnancy losses predicted the number of visits, after other variables had been taken into account. Married marital status was a positive predictor for the early initiation of care, after past pregnancies and risk score have been taken into account. PMID- 11428226 TI - The incidence of injury with the British Antarctic Survey, 1986-1995. AB - Medical consultations recorded by the British Antarctic Survey between 1986-1995 were analysed. A total of 3683 new consultations were recorded, an incidence of 2193.7/1000/year, with a significantly higher incidence on the bases compared to the ships. Injuries comprised 30.2% of consultations, an incidence of 661.7/1000/year. The bases had a significantly higher incidence of injuries. Work accounted for 47.0% and recreation 30.6% of all injuries. PMID- 11428227 TI - From "trailing edge" to "leading edge": an employer's diabetes approach. PMID- 11428228 TI - Challenges to implementing employee wellness programs. PMID- 11428229 TI - Are we missing the grade on report cards? PMID- 11428230 TI - Health plan incentives and quality improvement: do employers support them? PMID- 11428231 TI - Getting optimal bang for the buck in diabetes services. PMID- 11428232 TI - Efficiently and effectively educating patients with diabetes. PMID- 11428233 TI - Transfer of health data in the post-HIPAA era. PMID- 11428234 TI - Risk of enteric illness associated with travel: a case review of gastroenteritis among Canadian travellers: January to April, 2000. PMID- 11428235 TI - Outbreak of Ebola hemorrhagic fever, Uganda, August 2000-January 2001. PMID- 11428236 TI - [Pneumococci: a little known danger]. PMID- 11428237 TI - [Pneumococcal pneumonia in everyday practice. Why prevent it?]. PMID- 11428238 TI - [Pneumococcal pneumonia: the particular case of aged patients in institutions]. PMID- 11428239 TI - [Pneumococcal infections and risk factors: in whom should we prevent?]. PMID- 11428240 TI - Inhibition of cytomegalovirus immediate early gene expression: a therapeutic option? AB - The replication cycle of the human cytomegalovirus (HCMV) is characterized by the expression of immediate early (IE), early (E), and late (L) gene regions. Current antiviral strategies are directed against the viral DNA polymerase expressed during the early phase of infection. The regulation of the IE-1 and IE-2 gene expression is the key to latency and active replication due to their transactivating and repressing functions. There is growing evidence that the pathogenic features of HCMV are largely due to the abilities of IE-1 and IE-2 to transactivate cellular genes. Consequently, current drugs used to inhibit HCMV infection would have no impact on IE-1 and IE-2-induced effects that are produced before the early phase. Moreover, when HCMV DNA replication is inhibited, IE gene products accumulate in infected cells causing disturbances of host cell functions. This review summarizes the biological functions of HCMV-IE gene expression, their relevance in pathogenesis, as well as efforts to develop novel treatment strategies directed against HCMV-IE expression. PMID- 11428241 TI - Position statement: global neuraminidase inhibitor susceptibility network. PMID- 11428242 TI - Innate gastrointestinal immunity: characterization of broadly active viral inhibitors. AB - Innate viral inhibitors that are broadly active have been characterized in the serum and the nervous system, but incompletely characterized in the gastrointestinal (GI) tract. GI preparations from porcine gastric mucosa, mouse intestine, and in neuramide (a pharmaceutical product), were examined for broad antiviral activity, molecular size and mechanism of action for comparison with the previously characterized, innate inhibitors in the serum and nervous system. The GI inhibitors were found to be active in high titers against RNA and DNA viruses, resistant to proteolysis, glycolysis, lipid extraction and possessed differing mechanisms of action. The mouse intestinal inhibitor prevented virus attachment to cells, and neuramide acted at an early post-attachment stage of virus multiplication. The porcine mucosal inhibitor acted as late as 6 h after initiation of the multiplication cycle. These broadly active GI inhibitors differed from the previously described serum inhibitor (UTI beta) high density lipoproteins (HDL) and the nervous system (NS) inhibitor by being smaller (600 +/ 400 kDa) and resistant to proteinase K, glycosidases and organic solvents. The mouse intestinal inhibitor acts similarly to UTI beta and NS inhibitor by preventing attachment of virus to the cells. In comparison, the neuramide and the porcine mucosal inhibitor, like HDL, acted after attachment to the target cells. The innate nonspecific, broadly-active virus inhibitors, based on high titers and location, are considered important initial immune defense mechanisms against viral infections and thus potentially useful in medical applications. PMID- 11428243 TI - In vitro antiviral activity of the anthraquinone chrysophanic acid against poliovirus. AB - Chrysophanic acid (1,8-dihydroxy-3-methylanthraquinone), isolated from the Australian Aboriginal medicinal plant Dianella longifolia, has been found to inhibit the replication of poliovirus types 2 and 3 (Picornaviridae) in vitro. The compound inhibited poliovirus-induced cytopathic effects in BGM (Buffalo green monkey) kidney cells at a 50% effective concentration of 0.21 and 0.02 microgram/ml for poliovirus types 2 and 3, respectively. The compound inhibited an early stage in the viral replication cycle, but did not have an irreversible virucidal effect on poliovirus particles. Chrysophanic acid did not have significant antiviral activity against five other viruses tested: Coxsackievirus types A21 and B4, human rhinovirus type 2 (Picornaviridae), and the enveloped viruses Ross River virus (Togaviridae) and herpes simplex virus type 1 (Herpesviridae). Four structurally-related anthraquinones--rhein, 1,8 dihydroxyanthraquinone, emodin and aloe-emodin were also tested for activity against poliovirus type 3. None of the four compounds was as active as chrysophanic acid against the virus. The results suggested that two hydrophobic positions on the chrysophanic acid molecule (C-6 and the methyl group attached to C-3) were important for the compound's activity against poliovirus. PMID- 11428244 TI - Inhibition of murine cytomegalovirus and human cytomegalovirus by a novel non nucleosidic compound in vivo. AB - Novel non-nucleosidic compounds have recently been identified as potent inhibitors of the human cytomegalovirus (HCMV) and murine cytomegalovirus (MCMV) in vitro. We have now investigated the antiviral activity of these compounds in MCMV-infected NOD/LtSz-scid/j mice that lack functional T, B and, in contrast to C.B-17/Icr scid/scid mice, natural killer cells, and represent a novel model for cytomegalovirus infection in immunocompromised hosts. BAY 38-4766 (3-hydroxy-2,2 dimethyl-N-[4(([5-(dimethylamino)-1-naphthyl]sulfonyl)amino)- phenyl]propanamide) was identified as the most potent representative of this class of antiviral compounds. Per os administration of BAY 38-4766 at dosages > or = 10 mg/kg body weight led to antiviral effects that were comparable to ganciclovir 9-(1,3 dihydroxy-2-propoxymethyl)-guanine (Cymevene) as measured by survival and levels of viral DNA in organs of infected mice. In order to assess the anti-HCMV activity of BAY 38-4766 in vivo, we used a model, in which HCMV-infected human cells were entrapped in hollow fibers and subsequently transplanted into immunodeficient mice. Using this model, we demonstrated antiviral activity of BAY 38-4766 similar to that of ganciclovir. We conclude that BAY 38-4766 shows potential as an anti-HCMV drug. PMID- 11428245 TI - Enhancing readiness-to-change substance abuse in persons with schizophrenia. A four-session motivation-based intervention. AB - This article describes a four-session intervention designed for persons with co occurring substance abuse and schizophrenia-spectrum disorders, to be administered as an add-on module to supplement ongoing mental health treatment in an outpatient setting. The intervention targets those dually diagnosed individuals with low readiness-to-change as indicated by current use, and/or low level of engagement in treatment for substance abuse. The intervention is designed to increase problem recognition, to enhance motivation to change maladaptive patterns of substance use, and to facilitate engagement in substance abuse treatment. To achieve these goals, the authors have adopted constructs from the Transtheoretical Model of Change, the authors used principles of motivational and harm reduction interventions, and tailored them to the target population. PMID- 11428246 TI - An ABAC comparison of two intensive interventions for food refusal. AB - An ABAC comparison of two treatment packages for food refusal, physical guidance and nonremoval of the spoon, was conducted with two children with limited food acceptance. Both of these treatment packages included prevention of escape from presented food. Subsequent to baseline, one of the two treatment packages was implemented for each child. The treatment packages were implemented ABAC for one child and ACAB for the other child. Once the percentage of bites accepted had increased to at least 75% with the initial exposure to a treatment package, that treatment was withdrawn with a subsequent exposure to the second treatment package. The results indicated that both treatment packages were effective in establishing food acceptance. Also, initial exposure to either of the two treatment packages facilitated acquisition of food acceptance during the second exposure. Parental preference of the treatment package may have been influenced by the order of exposure to the treatment conditions. PMID- 11428247 TI - Stereotyping among providers and consumers of public mental health services. The role of perceived group variability. AB - The authors examine stigmatization and mental illness, focusing on the role of perceived group variability in stereotype use. Consumers' and providers' in-group and out-group stereotypes were assessed. Although providers had extensive experience, they judged consumers more stereotypically and just as negatively as did the consumers themselves. Consumers' education and involvement in services were weakly predictive of more stereotypic, less variable, and more negative views of providers, whereas providers' education and involvement in services predicted more stereotypic but also more variable views of both groups. Perceived group stereotypicality predicted more stereotypic judgments of individuals, whereas perceived variability predicted less confidence in judgments. Because providers perceived greater variability, they were less confident in applying the stereotype to individuals. We suggest that increasing perceptions of the variability among consumers may lead to more sensitive use of diagnostic criteria, more individualized treatment, and a decrease in the negative effects of stigmatization. PMID- 11428248 TI - Food selectivity and problem behavior in children with developmental disabilities. Analysis and intervention. AB - Excessive food selectivity typifies some children with developmental disabilities. We conducted functional analyses to determine the controlling variables for problem behavior that accompanied food selectivity and analyzed the role of establishing operations in ameliorating food selectivity. Specifically, we studied the differential effects on intervention efficacy of an individual's having or not having access to preferred food items prior to an intervention that involved the presence versus absence of a positive reinforcement contingency applied to food consumption. Participants displayed significantly more problem behavior during the nonpreferred-foods condition. Participants consumed nonpreferred target food items only when prior access to preferred foods was limited and a positive reinforcement contingency was implemented. Functional analysis suggested that problem behavior was maintained by negative reinforcement. Intervention data suggested that establishing operations increased the efficacy of the contingency-based intervention. The implications of applying this intervention in the community were discussed as were the relative merits of stimulus fading versus escape extinction intervention strategies. PMID- 11428249 TI - Evaluation of a brief intervention for increasing seat belt use on a college campus. AB - The authors evaluated a brief intervention for increasing seat belt use among the front seat occupants of cars at a junior college, in a jurisdiction with a mandatory belt use law. The intervention included public posting of performance feedback and distribution of an informational flyer to cars in target parking lot. Feedback was the display of the proportion of drivers observed wearing seat belts on the previous observation day. Seat belt use among drivers increased from 64% during the baseline phase to 71% during the intervention phase. Seat belt use among front passengers increased from 49% during the baseline phase to 67% during the intervention phase. In both cases, seat belt use at follow-up was comparable to seat belt use during the intervention phase, although a trend toward decreasing belt use was noted. Also found was higher seat belt use among females as compared with males irrespective of their front seat occupant status (driver or passenger). Effects of the intervention are discussed in the context of increasing seat belt use in a hardcore nonuser population of predominantly young adults. PMID- 11428250 TI - Occupational health in European member states: a road to organizational health. AB - After an introduction, giving a short historical perspective on European Health and Safety legislation, the first experiences and opportunities for occupational physicians regarding international co-operation are described, with the first result in a form of a Position Paper on Occupational Health, adopted by the Standing Committee of European Doctors and presented to the European Commission. Since the ultimate shared goal for occupational physicians should be to contribute to the promotion of healthy workforce in safe and sound working conditions, the characteristics of a healthy organization are described with the possible activities to reach these characteristics, with particular attention drawn to the role of different disciplines. The growing importance of the management responsibility for health and safety in their enterprises is stressed and the possible benefits elucidated. The implications and consequences of such an extensive and ambitious program are pointed out with special regard to the challenge to contribute pro-actively to ensuring a healthy workforce in safe and sound working conditions for the benefit of employees, hereby clarifying the role and improving the general perception and position of occupational physicians. The importance of speaking with one voice and sharing ideas with colleagues, employers, employees and governments is essential, if the mission "to contribute to the realization of a healthy workforce in safe and sound working conditions" is to be reached. PMID- 11428251 TI - Current trends, examples of regulations and practical approaches to occupational health services in the United Kingdom. AB - Occupational health services in the United Kingdom are evolving from the traditional approach using doctor and nurses to provide clinical care at the worksite for any medical ailment, to multidisciplinary occupational health practitioners focussing on the prevention of ill-health from workplace factors. Nevertheless, there continues to be an artificial divide between safety departments and occupational health departments within the same organisation. Many occupational health services focus on the need to comply with the requirements of health and safety legislation. In the UK, these include the Health and Safety at Work, etc. Act of 1974, the Control of Substances Hazardous to Health, the 1994 regulations, and a newer legislation based on the European Union Directives. A practical approach to providing occupational health cover has been the development of occupational health departments within the public healthcare sector, private occupational health service providers, and independent consultants. These are some similarities between the UK situation and other countries in the models used for providing occupational health care. The appropriate model for any country would depend on their perceived needs, resources, industries and hazards. PMID- 11428252 TI - Multidisciplinary model of occupational health services. Medical and non-medical aspects of occupational health. AB - Since 1950, the International Labour Organization (ILO) and the World Health Organization (WHO) have had a common definition of occupational health. The definition was adopted by the Joint ILO/WHO Committee on Occupational Health at its First Session (1950) and revised at its 12th Session (Geneva, November 1995). Occupational health should aim to promote and maintain the highest degree of physical, mental and social well-being of workers in all occupations; to prevent amongst workers the departure from health caused by their working conditions; to protect workers in their employment from risk resulting from factors adverse to health; to place and maintain workers in an occupational environment adapted to their physiological and psychological capabilities; in summary, to adapt work to the workers and each worker to his or her job. According to the ILO Convention No. 161/1985 (1) and the 1996 WHO Global Strategy on Occupational Health for All, to protect the worker against sickness, disease and injury arising out of his or her employment, the establishing of occupational health services for all workers is recommended. The Convention as well as the Strategy emphasize the importance of multidisciplinary approach and multisectoral collaboration. It is evident that during the last decades of the 20th century, the concept of occupational medicine and occupational health has been changed. Occupational health services mean services entrusted with essentially preventive functions. Is there still a room for medical services provided by occupational physicians? The dominance of medical professionals seems to disappear in the modern multidisciplinary model of Occupational Health Services. PMID- 11428253 TI - WORKSAFE IOWA Occupational Medicine Associates Network: a university--community partnership in occupational safety and health. AB - The WORKSAFE IOWA Occupational Medicine Associates Network is a unique health care model for dispensing regional occupational medicine services in the state of Iowa, USA. WORKSAFE IOWA is an educational, informational, and consultation service of the College of Public Health at The University of Iowa. WORKSAFE IOWA includes a fee-for-service industrial hygiene program, the Occupational Medicine Associates Network, and educational and informational services. The Associates Network provides education, information, and consultation in exchange for an annual fee paid by each Associate to the university-based network. The Associates clinics provide comprehensive occupational medicine services to up to 1,500 clients through 66,000 annual clinic visits in their respective communities. The Associates Network has been financially self-sustaining over a period of 10 years, and has proven to provide valuable services to the community-based Associates, and to provide excellent training opportunities for the University students in several occupational safety and health disciplines. PMID- 11428254 TI - Good practice in health, environment and safety management in enterprise. AB - Good practice in health, environment and safety management in enterprise (GP HESME) is a process that aims at continuous improvement in health, environment and safety performance, involving all stakeholders within and outside the enterprise. This WHO program is supported by other international organizations, and the declaration of Ministers of Health and Ministers of Environment adopted in 1999. The basic issues of the GP HESME concept are presented as well as its prerequisites, benefits and participants. The key partners in GP HESME are employers and their organizations, representatives of employees, governmental agencies, local authorities, financial and insurance institutions, occupational health services, environmental and social services, associations of professionals, research and training institutions. The HESME system is intended to function at different levels: international, national, local community, and enterprise settings. The lists of expected benefits for each group of stakeholders are discussed. Evaluation of GP HESME is based on the criteria and indicators, the most important of them are briefly presented. PMID- 11428255 TI - Legal regulations on occupational health system in Poland. AB - Occupational health care has a long tradition in Poland. It has evolved from the industrial health care system established in the post-war period to a modern system defined in the Occupational Health Services Act of 1997. When the process of political and economic transformation began in Poland in 1989, the reforms of the workers' health care became inevitable. The process of introducing and implementing new legal regulations comprised three phases: 1. Introduction of amendments to the Polish Labor Code, according to which employers are now committed to provide their workers with occupational health services (OHS) appropriate to given working conditions. 2. Incorporation into the 1991 Health Care Institutions Act the provisions which allow for the establishment of alternative non-public structures responsible for the health care of the working population. 3. Adoption of the Occupational Health Services Act in 1997, the most important law providing grounds for establishing a most comprehensive system of occupational health services. The Occupational Health Services Act introduces, for the first time, the concept of the OHS system that encompasses the whole working population. The Act outlines a broad range of OHS competences; defines individual tasks; and provides the up-to-date definition of the OHS structure with its two organizational levels, consisting of primary and regional occupational health centers. In addition, the Act specifies the sources of finance for the accomplishment of the defined tasks. Pursuant to the provisions of the Act, OHS units co-operate with employers and employees, bodies supervising working conditions and other organizations involved in occupational health that become their partners in activities aimed at protecting and promoting health of workers. In order to further develop the workers' health care system its constant adaptation to changing conditions is absolutely necessary through: developing modern training for specialists in the field of occupational medicine; establishing an efficient system for controlling the quality of services provided by the occupational health care centers; and implementing more effective forms of health protection and promotion at workplace, especially for those self-employed. PMID- 11428256 TI - Summary report on the workshop "Occupational health from the perspective of future accession of Poland to the European Union" Lodz, Poland, 30 June-1 July, 2000. PMID- 11428257 TI - The role of the School of Public Health at the Nofer Institute of Occupational Medicine in multidisciplinary training of physicians in occupational medicine. AB - The School of Public Health was opened on 30 June 1992 as an integral part of the Nofer Institute of Occupational Medicine. The late Director General of the Institute, Professor Janusz A. Indulski (1930-1999) was its initiator and founder. The School of Public Health is a scientific, research and didactic unit of the Nofer Institute. It offers post-graduate studies in the areas of occupational medicine, work hygiene, public health, environmental health, organization and management of health care and health promotion. The opportunity for multidisciplinary education of specialists in occupational medicine lies in the fact that the didactic process organized by the School is based on the knowledge, experience and skills of the Institutes' research workers. Lectures are provided by 100 persons, including 22 professors, 6 assistant professors and over 50 doctors. The didactic team also includes outstanding national and international experts (WHO, OECD, IAAE, etc.). During the last five years, the number of students increased from 1200 in 1996 to over 2200 by the end of 2000. PMID- 11428258 TI - Training of occupational physicians in The Netherlands with regard to occupational health services delivered to the population. AB - In the Netherlands, the need for a basically new approach to education and training in occupational medicine was felt by professionals, students, schools and occupational health services (OHS) in the early 1990s. After an inventory of the problems and shortcomings of the traditional curriculum, the Netherlands School of Occupational Health defined the framework for a new curriculum. In this article the background, principles and structure of the new curriculum are described. Three principles shape the curriculum: the needs of OHS; professional standards; and the state-of-the-art. The characteristics of the new curriculum are: interaction between theory and practice; students' self-management of the learning process; co-makership with OHS; and multidisciplinarity. The curriculum consists of a course/theory and a practical part. Most of the theoretical part is presented to so called core group of 12 students, which is to be maintained during the full course period of 4 years. The adage for the practical part to be spent in a certified OHS institution is: "the best teaching OHS are learning OHS". In 1999, the first group of students entered the renewed curriculum. First impressions of the experience gained are presented. PMID- 11428259 TI - The competences postulated as requirements for occupational medicine training in Europe as viewed by Polish specialists. AB - A questionnaire has been developed and disseminated in several European countries to determine common key competences required of occupational medicine specialists. The questionnaire contained 115 subjects relating to eight fields of activity of an occupational medicine physician (occupational hazards to health, assessment of disability and fitness for work, communications, research methods, management, environmental medicine, occupational health law and ethics, and health promotion). Items in each part were classified into three categories: knowledge, skills and experience. For each of the subjects respondents were asked to allocate a score from 0 (not necessary) to 5 (most important or essential). In Poland the questionnaire was distributed among two groups of specialists: (1) chief administrators of occupational health services, and (2) relatively young occupational medicine physicians attending a specialist training. A comprehensive analysis of the completed questionnaires had three dimensions: (a) substantive (classification of the importance of particular key competences, as perceived by Polish specialists in occupational medicine); (b) personal (differences in opinions among occupational medicine physicians and an attempt to explain these differences in sociological terms); and (c) comparative (evaluation and interpretation of similarities and differences between two groups). PMID- 11428260 TI - Principles for shaping working conditions according to ILO conventions and EC directives. AB - The strategy for the improvement of occupational safety and health is presented. The role of the state and the workers in shaping working conditions has changed over the years, and this process is traced through ILO and EC documents. There must be a coherent up-to-date national policy in this field. Safety and the protection of the workers' health is perceived by the Community legislation not only in the terms of the worker's individual interest, but also from a social viewpoint, consisted in striving for general improvement of the work environment. The Community regulations aspire to satisfy the public interest, and directives set the maximum level of protection but also serve as minimum standards not infringing the more favourable national regulations. They define requirements that should already be met at the planning and designing stages. PMID- 11428261 TI - AUDIOBUS--the first Polish audiological mobile unit. PMID- 11428262 TI - Occupational medicine in the Internet--educational Internet service. PMID- 11428263 TI - Electrophysiological approaches to the study of neuronal exocytosis and synaptic vesicle dynamics. PMID- 11428264 TI - Input-output functions of mammalian motoneurons. AB - Our intent in this review was to consider the relationship between the biophysical properties of motoneurons and the mechanisms by which they transduce the synaptic inputs they receive into changes in their firing rates. Our emphasis has been on experimental results obtained over the past twenty years, which have shown that motoneurons are just as complex and interesting as other central neurons. This work has shown that motoneurons are endowed with a rich complement of active dendritic conductances, and flexible control of both somatic and dendritic channels by endogenous neuromodulators. Although this new information requires some revision of the simple view of motoneuron input-output properties that was prevalent in the early 1980's (see sections 2.3 and 2.10), the basic aspects of synaptic transduction by motoneurons can still be captured by a relatively simple input-output model (see section 2.3, equations 1-3). It remains valid to describe motoneuron recruitment as a product of the total synaptic current delivered to the soma, the effective input resistance of the motoneuron and the somatic voltage threshold for spike initiation (equations 1 and 2). However, because of the presence of active channels activated in the subthreshold range, both the delivery of synaptic current and the effective input resistance depend upon membrane potential. In addition, activation of metabotropic receptors by achetylcholine, glutamate, noradrenaline, serotonin, substance P and thyrotropin releasing factor (TRH) can alter the properties of various voltage- and calcium-sensitive channels and thereby affect synaptic current delivery and input resistance. Once motoneurons are activated, their steady-state rate of repetitive discharge is linearly related to the amount of injected or synaptic current reaching the soma (equation 3). However, the slope of this relation, the minimum discharge rate and the threshold current for repetitive discharge are all subject to neuromodulatory control. There are still a number of unresolved issues concerning the control of motoneuron discharge by synaptic inputs. Under dynamic conditions, when synaptic input is rapidly changing, time- and activity-dependent changes in the state of ionic channels will alter both synaptic current delivery to the spike-generating conductances and the relation between synaptic current and discharge rate. There is at present no general quantitative expression for motoneuron input-output properties under dynamic conditions. Even under steady state conditions, the biophysical mechanisms underlying the transfer of synaptic current from the dendrites to the soma are not well understood, due to the paucity of direct recordings from motoneuron dendrites. It seems likely that resolving these important issues will keep motoneuron afficiandoes well occupied during the next twenty years. PMID- 11428265 TI - Transport of proteins into mitochondria. AB - Most mitochondrial proteins are nuclear-encoded and synthesised as preproteins on polysomes in the cytosol. They must be targeted to and translocated into mitochondria. Newly synthesised preproteins interact with cytosolic factors until their recognition by receptors on the surface of mitochondria. Import into or across the outer membrane is mediated by a dynamic protein complex coined the translocase of the outer membrane (TOM). Preproteins that are imported into the matrix or inner membrane of mitochondria require the action of one of two translocation complexes of the inner membrane (TIMs). The import pathway of preproteins is predetermined by their intrinsic targeting and sorting signals. Energy input in the form of ATP and the electrical gradient across the inner membrane is required for protein translocation into mitochondria. Newly imported proteins may require molecular chaperones for their correct folding. PMID- 11428266 TI - [Long-term functional results in ossicular reconstruction with cartilage]. AB - Optimal hearing is one goal of otologic surgery. This study examines the long term functional success rate of cartilage as graft material for ossicular chain reconstruction in type III tympanoplasty. A retrospective review of seventy-nine patients having undergone type III tympanoplasty (53 with closed techniques and 26 with open techniques) was made. Lateral graft ("onlay") techniques were used in almost all cases. All operations were performed by one of the authors, a well established otologic surgeon. Seventy-four percent of the reconstructions were performed in an one-stage procedure. The mean period of follow-up was 54 months (range 1-17 years). The results of air-bone gap closure and complications of the surgery are presented. Our results show that a significant percentage of patients gain a long-term substantial improvement in their hearing with minimal risk. However, this hearing improvement tends to decline eventually in a progressive manner. A post-operative air-bone closure equal or < 30 dB was achieved in 92.2% of the all cases (n = 79) with a mean gain of 9.72 dB. Functional outcomes in closed techniques were slightly better than in open techniques. There were few complications. Perforation and retraction of the tympanic graft were the most common observed complications. In conclusion, cartilage is an available, costless, reliable and technically easy to use graft material for functional reconstructions of the ossicular chain. It is a well-tolerated element in middle ear space as well as by the tympanic membrane, it has a very low risk of extrusion and a good long-term functional viability. Therefore, update cartilage is an interesting option in functional restoration of the ossicular chain as graft placed on the stapes capitulum. PMID- 11428267 TI - [Auditory training with wide-band white noise]. AB - The auditory training with wide-band white noise is a methodology for the qualitative recovery of the hearing loss in people suffering from sensorineural hearing loss. It is based on the application of a wide-band modified noise which we will call "K-noise". The auditory trainer GAES 100 KT is a voice amplifier device with earphone outlets. It incorporates a wide-band noise generator which produces a noise that is a mixture of all the audibles frequencies. Therefore, it stimulates all the sensory cells in the organ of Corti, but from 1000 Hz it has a progressive fall towards the high pitch of 6 db per octave. This device consists of a series of controls which make possible to apply both the noise and the phonetic materials--texts and words lists that the therapist's voice presents--to either one of the ears or to both of them, in different intensities and for different periods of time. In this first article of our study of the auditory training with wide-band white noise we will review the different methods of hearing training and present not only the basic concepts on auditory training, but also the systematic we have followed to apply it. PMID- 11428268 TI - [Maxillary sinus hypoplasia]. AB - Maxillary sinus hypoplasia is rare, with an estimated prevalence of 1-5%. Out of the CT scans performed in sinusal patients between March 1998 and June 1999, we report on 4 isolated maxillary sinus hypoplasia, 4 maxillary sinus hypoplasia associated to concha bullosa, and 10 isolated conchae bullosas. All cases were evaluated by nasosinusal endoscopy and CT scan. Size, location and uni/bilateral presentation of concha bullosa is correlated to maxillary sinus hypoplasia presence, specially with regards to uncinate process presence, medial or lateral retraction. The pathogenesis of maxillary sinus hypoplasia is reviewed, and its relation to concha bullosa, evaluating how this could explain some cases of the so called chronic maxillary sinus atelectasia, as an acquired and progressive variant of maxillary sinus hypoplasia in adults. PMID- 11428269 TI - [Parapharyngeal abscess: perilaryngeal infrahyoid extension]. AB - Parapharyngeal abscesses are located between the skull base and the hyoid bone. Their infrahyoid extension is usually following the retropharyngeal space, but also can be through the medial aspect of the parapharyngeal space. Thus, surrounding the visceral fascia, they might invade the perilaryngeal and perihypopharyngeal spaces. This results in more significant airway compromise. Computed tomography is essential while locating and evaluating the extension of these infections, specially with regards to the great vessels. We report on 8 parapharyngeal abscesses with perilaryngeal and/or perihypopharyngeal extension. We review their clinical and radiological features, based on computed tomography, as well as their medical and surgical treatment. When medical therapy fails, as they are usually medial to the great vessels, it is possible to perform aspiration and drainage transorally, avoiding the need for cervicotomy, in selected cases. PMID- 11428270 TI - [Antibiotic prophylaxis in oncologic pharyngolaryngeal surgery: ceftriaxone versus clindamycin and gentamycin]. AB - There are many papers comparing two antibiotic protocols for the profilaxis of head and neck infections after laryngeal surgery. We present one prospective and randomised study in 60 patients comparing the efficacy of two protocols. The comparison was between ceftriaxone versus the association of clindamicyn and gentamicyn. In our database we included the risk factors for infection, the surgical approach, the duration of surgery and the patient characteristics. We observed an incidence of 28% of infection, with a 23.3% in the clindamicyn + gentamicyn group and a 33.3% in the ceftriaxone group. The differences between the two groups were not statistically significant. In this study we observed a small difference between the amount of alcohol comsuption, the effectiveness of the surgical drainage, the surgical approach and the presence of wound infection. The difference was not statistical significant due to the small group of patients. The profilaxis was adequate for the total laryngectomy and cordectomy group, with a higher incidence of wound infection in patients treated with a supraglottic laryngectomy. PMID- 11428271 TI - [Giant cholesteatoma with intact tympanic membrane]. AB - The cholesteatoma is characterized by Epidermic Keratinizated tissue in the middle ear and mastoid with capacity of migration and erosion of near-by structures. Commonly cholesteatoma is associated with chronic medium otitis, but in a few cases, tympanic membrana is integry and the diagnosis is made when appear complications. We report a case of giant cholesteatoma with intact tympanic membrana. The extension, silent period and evolution are evaluate. PMID- 11428272 TI - [Frontal ethmoid metastases of prostatic carcinoma. Report of one case and review of the literature]. AB - Prostatic metastases in the nose and paranasal sinuses are rare. Less than 100 cases have been reported in the literature. Kidney are the commonest site of primary tumour, followed by lung and breast. Only 10 cases have previously been reported in the world literature. Prostatic metastases have been mainly reported in the sphenoid sinus. This paper reports one case of metastases of prostatic carcinoma in the fronto-ethmoid sinus in a 72 years old male. The clinical picture includes acute fronto-ethmoid right sinusitis, severe exophthalmos and chemosis. The CT scan showed extensive soft tissue filling the maxillary, ethmoid cells, sphenoid and frontal right sinuses, with subdural abscess. Biopsies from the fronto-ethmoid mass showed infiltration by adenocarcinoma with positive immunostaining for prostatic specific antigen. We also review the literature about metastases involving the nose and paranasal sinuses. PMID- 11428273 TI - [Benign sinusal mucor colonization in association with septal deviation]. AB - Mucormycosis is a well recognised opportunistic infection caused by saprophytic fungi of the Mucoraceae family. Usually, the upper aerodigestive tract is the portal of entry. Patients who are debilitated or inmunosuppressed might present with these infections, although sometimes can happen in patients with good health. We present a case of benign mucor colonitation at the sinus maxillary associated a deviated nasal septum. PMID- 11428274 TI - [Lemierre's syndrome: septic thrombophlebitis of the internal jugular vein secondary to acute amygdalitis]. AB - Lemierre's syndrome is an uncommon clinical entity. It consists of an acute oropharyngeal infection, with secondary septic thrombophlebitis of the internal jugular vein frequently complicated with multiple metastatic infections. It is generally caused by anaerobic Gram-negative organisms. Although it is rarely reported in the antibiotic era, this disease must be taken into account because it is a potentially life-threatening infection requiring a specific and early treatment. We report a case of Lemierre's syndrome managed in our hospital. PMID- 11428275 TI - [Commentaries on the complication of vestibular diseases]. AB - Handicap of the patient with vestibular disease can be measured by means of scales and questionnaires. The utility of a questionnaire developed and validated in Spanish language is commented (CEMPV). The CEMPV consists of 46 questions distributed in three scales (emotional, functional and organic) whose obtained scores are related to diverse degrees of handicap (global, emotional, functional and organic). The CEMPV is self-employed and evaluates the evolution of the patient and allows to accredit the results obtained between different investigators. PMID- 11428276 TI - [Otosclerosis and Van der Hoeven's syndrome: a contribution]. AB - Morphological and microchemical changes that effect to the otosclerotic stape in the Van der Hoeve's syndrome were examined with a scanning electron microscope equipped with an energy dispersive X-ray fluorescence. Using the Ca/P ratio as criterion--measured by the characteristic x-ray fluorescence--it was shown that the Van der Hoeve stape had a higher Ca/P ratio (2.6:1) as compared to the normal stape (2:1). The Van der Hoeve's syndrome lesions as poorly mineralized, with low calcium salt and apparent increase of phosphates. This finding indicates a possible change from hydroxyapatite (or apatite) to brushite, which imply an acidification of bone. PMID- 11428278 TI - Each time bears its own burden. PMID- 11428277 TI - [Importance of impedance audiometry on infant hearing screening test with otoacoustic emissions]. AB - The target of this paper is to evaluate the importance of Impedanciometry in a protocol with transient evoked otoacoustic emissions on high risk infant hearing screening. We used tympanometry and stapedious reflex only when otoacoustic emissions became altered. This method try to decrease the impact in this test caused for middle ear diseases. We realized an Impedanciometry study in all children with abnormal otoacoustic responses and then, we obtained normal otoacoustic responses when tympanometry normalize. In the other hand, the time of test wasn't too large, between 15-20 minutes for child. The analysis of specificity and positive predictive value, of otoacoustic emissions without Impedanciometry was 89% and 45% respectively, however the same values with Impedanciometry was 96% and 75% respectively. In our opinion, high frequency of middle ear diseases in children, and our results, justify the introduction of Impedanciometry in a complete protocol of hearing screening with Otoacoustic Emissions, preventing appearance of false positive caused by these diseases. PMID- 11428279 TI - Radiologic changes of ischial tuberosity in ankylosing spondylitis. AB - The objective of the paper was to emphasize the meaning of involvement of ischial tuberosity in ankylosing spondylitis, enriching the clinical picture of the disease and pointing to ankylosing spondylitis, especially when sacroiliitis is absent or less developed. Radiologic examination of ischial tuberosity was performed in 68 patients with different developmental stages of ankylosing spondylitis. There were 66 (97%) men and two (3%) women, aged 30 to 56 years. Anteroposterior radiographs of the pelvis were performed in all patients and independently assessed by two radiologists. The radiologic changes of ischial tuberosity were classified into four stages: 1) stage of minimal changes, 2) stage of destructive changes, 3) stage of reconstructive changes, and 4) stage of ossification of tendon fibers in the form of rumpled tufts. Radiologic changes of enthesitis were found on ischial tuberosity in 31 patients (45.5%, p < 0.01). Stage 1 or minimal changes were found in two (6.5%); stage 2 or destructive changes in 13 (42.0%); stage 3 or reconstructive changes in 11 (35.5%) and stage 4 or "tufts phenomenon" in five (16.0%) patients. The classification of changes is useful for the assessment of disease range and progression as well as for the diagnosis when sacroiliitis is obscurely developed and other radiologic signs are not present. PMID- 11428280 TI - Transfusion medicine in the new millennium. AB - Blood is a tissue like all other tissues, however, it has always had a special meaning for man. Blood used to be attributed characteristics of the man whose body it was circulating through. Attempts were made to influence the person's character, properties, or disease, such as strength, old age and illness, by the bloodletting or blood administration. Blood was for centuries considered an elixir of life, and was ascribed various meanings at different times, e.g., mystic, religious, symbolic, racial, patriotic, biological, medical, scientific, industrial, and economic. Nowadays, the meaning of blood is most pronounced in the science, medicine, and economy. PMID- 11428281 TI - My term of office at the Academy of Medical Sciences of Croatia 1963-1983. A contribution to the history of the academy. PMID- 11428282 TI - Laparoscopic management of infantile hypertrophic pyloric stenosis. AB - Infantile hypertrophic pyloric stenosis is a common problem in pediatric surgery. Conventional management by the upper laparotomy was the method of choice over the last few decades. Advanced minimally invasive surgery allows successful endoscopic management of this entity too. We report on our initial experience with endoscopic surgery in the treatment of infantile hypertrophic pyloric stenosis with respect to some technical details. The operative procedure was well tolerated by the infant. After a short and uneventful postoperative course, the infant regained eating habits and was discharged from the hospital on the fifth postoperative day. Our favourable initial experience suggests that laparoscopic pyloromyotomy could be a safe and efficient alternative to the open surgery. PMID- 11428283 TI - Congenital tracheal stenosis treated with dilatation. AB - Congenital tracheal stenosis is a rare anomaly characterized by a broad spectrum of variations. Affected segments differ in the grade extent of stenosis, from thin webs to more severe stenoses such as short-segmental, funnel-like, or long segment tracheal stenosis. A 5-month-old infant with persistent congenital stridor and acute respiratory distress is presented. Radiographic and tracheobronchoscopic investigations confirmed the diagnosis of congenital tracheal stenosis without associated vascular, pulmonary or gastrointestinal anomalies. The stenosis was of a fibrous nature and segmental morphology affecting the initial tracheal section. Dilatation was performed by a rigid tracheobronchoscope with favorable outcome. The reported case was classified into class 1 lesions, which are associated with low mortality and good prognosis. Dilatation performed with a rigid bronchoscope proved to be a rational therapeutic approach in this case. The possible diagnostic and other therapeutic approaches to congenital tracheal stenosis are also discussed. PMID- 11428284 TI - Diagnostic accuracy of transesophageal echocardiography for detection of atrial masses. AB - The aim of this study was to assess the diagnostic accuracy of transthoracic (TTE) and transesophageal echocardiography (TEE) for the detection of atrial masses. The authors' own experiences with the use of TTE and TEE images in the assessment of atrial masses are reported. These masses included tumors, thrombi, and valvular vegetations. The study groups consisted of 14 consecutive patients (7 women an 7 men), age range 24-72 (mean age 56.6 < +13.4) years. Eleven patients had left atrial tumor, two patients had atrial thrombi, and one patient had vegetation in left atrium detected with TEE. Eight patients had left atrial myoma, two patients had right atrial myxoma, and one patient had right atrial leiomyosarcoma. There was no false negative and no false positive TEE diagnosis, yielding a 100% sensitivity and specificity of TEE in detecting atrial masses. TEE detected atrial masses in six (43%) patients, provided poor images in five (36%) patients, and failed to reveal atrial masses in three (21%) patients. The TEE diagnosis was confirmed by surgery and pathohistology in all patients. The ability of TEE to visualize both atria with great diagnostic accuracy makes it a very valuable procedure in the assessment of atrial masses. PMID- 11428285 TI - Clinical, epidemiological and epizootic features of Q fever in the northern coastal part of Croatia from 1989 to 1998. AB - Although Q fever is endemic in the northern coastal part of Croatia, it usually occurs sporadically. Analysis of 58 patients hospitalized for Q fever during the last 10-year period (1989-1998) revealed some differences in the clinical manifestation compared to a previous study (1954-1977). Most cases of Q fever (N = 55; 91%), presented with pneumonia, but no rash was noticed, compared to 46% of patients with Q fever developing exanthema in the previous study. The previously observed high seropositivity to Coxiella burnetii among domestic animals was confirmed in this study. A two-peak seasonal distribution of Q fever observed in 1991 was connected with the imported Russian sort of sheep with special biology of delivery. The clinical outcome was favorable for all patients, since no complications or chronic forms of the disease were recorded. Disproportion between the number of registered and hospitalized patients, including a number of asymptomatic and several undiagnosed or misdiagnosed infections, leads to a conclusion that the real number of persons infected with Coxiella burnetii in the area is several times higher. PMID- 11428286 TI - [The importance to nineteenth century hospital medicine of an anatomic specimen of severely traumatized vertebrae. Scientific history at the Museum of Pathologic Anatomy in the Hospital of Trieste]. AB - The museum of pathological anatomy, established in Trieste in 1847, a few years after the opening of the general hospital, keeps a sole pathological finding of "lethal vertebral trauma" dated 1903 and prepared by one of the fathers of Trieste's pathological anatomy, Enrico Ferrari (1874-1970). This museum object recalls a clinical sensitivity in traumatology going back to Simone Pertot (1845 1907), the first pathologist of Trieste, but also the clinical observations of one of Billroth's disciples, the triestine Arthur Menzel (1844-1878), head of the i.v. Surgery Division. Giving this museum object its historical scientific genesis means answering the following questions: Why does the pathologist provide clinical medicine with such a preparation? In other words, what pragmatic meaning does it have? What is its function? In the practical clinical activity of the above-mentioned hospital's medicine, the state of disease finds its meaning in concepts like diagnosis and prognosis and, therefore, the same epicritic function of the pathologic sample representing vertebral trauma. Its "clinical" meaning is in the pathologist's and clinician's ability at elaborating a pathognomonic sign which is able to put forth, at the patient's bedside, the physiopathological laws determining the seriousness of the trauma taking place in the organism. PMID- 11428287 TI - [Demonstration of TT virus in liver tissue fixed in formalin and embedded in paraffin]. AB - INTRODUCTION: TT virus has been recently isolated in Japan in patients with acute and chronic non-A/non-G hepatitis. Its possible etiopathogenetic role in causing hepatitis has been initially taken in consideration. On the contrary, more recent studies deny the importance of TT virus in causing liver damage. Most of the studies are based on serological data or on viral detection from frozen liver tissue. AIM OF THE STUDY: In the present paper we describe a method to detect viral genome from formalin-fixed and paraffin-embedded liver tissue. MATERIALS AND METHODS: Twelve needle biopsies from liver were studied. Six cases were selected on the basis of serological negativity for HBV and HCV markers. Five cases of HCV-related chronic hepatitis and one HCV- and HIV-positive intravenous drug abuser were also included. All patients underwent liver biopsy, performed with a 14-G needle. Liver specimens were formalin-fixed and paraffin-embedded as routine. From each block, sections were cut and stained for histopathologic examination. Additional 5 microns sections were employed to extract DNA for nested PCR. RESULTS: In 2 of 12 cases studied, TT virus genome was found. In both cases the presence of viral DNA was confirmed by sequencing. Both patients were male. The first patient was a 39-year-old HIV- and HCV-positive intravenous drug abuser. The second patient was a 60-year-old heavy alcohol drinker. In both cases the presence of TT virus apparently did not affect the histological picture. CONCLUSION: It is possible to detect TT virus genome from formalin-fixed and paraffin-embedded tissue. This method offers the possibility to perform retrospective studies. PMID- 11428288 TI - [Cytogenetic analysis of 19 renal cell tumors]. AB - To further evaluate the role of cytogenetic analysis we studied 19 cases of renal neoplasms. Specific chromosomal aberrations have been demonstrated associated with different histologic types. Particularly, clear cell renal cancers were associated with deletions of the short arm of chromosome 3 and papillary renal cell cancers demonstrated multiple trisomies, and chromophobe cancers and oncocytomas were characterized by loss of whole chromosomes. The utility of cytogenetics as a tool to define the pathological spectrum of renal cell neoplasms is stressed. PMID- 11428289 TI - [Malignant phyllode tumor of the breast with features of intraductal carcinoma]. AB - Malignant phyllode tumor is a rare biphasic breast tumor consisting of a malignant mesenchymal component and an epithelial component that is usually benign. We report an unusual case of a malignant phyllode tumor of the breast with neoplastic features of both the epithelial and stromal components. The patient was a 39-year-old woman with family history for breast carcinoma. Grossly, the excised tumor was a 9 x 7 x 5.5 cm gray lobulated mass with infiltrative margins and necrotic-hemorrhagic areas. Histologically the tumor consisted mainly of neoplastic mesenchyme with non invasive comedo, cribriform and micropapillary features of the ducts. Three months after the excision of the neoplastic mass, the patient developed an infiltrating ductal carcinoma of the opposite breast. Hereditary and bilateral tumors are commonly associated with germline mutations. Tissue from both neoplasms however did not express either BRCA1 or BRCA2 mutations. PMID- 11428290 TI - [Villograndular adenocarcinoma of uterine cervix: a case report]. AB - We report a case of villoglandular adenocarcinoma of the uterine cervix, discovered in a 69-year-old woman. The patient underwent a radical Wertheim hysterectomy and bilateral pelvic lymphoadenectomy and is alive and well, with no evidence of recurrent disease after 5 years from diagnosis. 13 reports totaling 70 patients with diagnoses of villoglandular adenocarcinoma have been published. The patients ages ranged from 22 to 61 years. Well-differentiated villoglandular adenocarcinoma of the uterine cervix has been recently recognized and described as a distinctive histologic entity which develops in young women. It is possible that cases of villoglandular adenocarcinoma in older women, as our case, were previously considered to represent different histologic types of adenocarcinoma. The differential diagnosis of villoglandular adenocarcinoma is important for prognostic and treatment reasons. PMID- 11428291 TI - [Synovial chondromatosis presenting as soft tissue mass]. AB - Primary synovial chondromatosis is a rare condition characterized by the formation of hyaline cartilaginous nodules; pathogenesis is not fully understood. A soft tissue mass is an even more rare presentation of synovial chondromatosis. Histologically the condition shows some atypical features that suggest a malignant process. We describe a case of primary synovial chondromatosis presenting as soft tissue mass. PMID- 11428292 TI - [Aneurysmatic fibrous histiocytoma: case report and reivew of the literature]. AB - A case of aneurysmal fibrous histiocytoma is described. The patient is a 26-year old man with a reddish nodule on the back, recently presenting a volume increase. The tumor was composed of fascicles of short spindle cells, histiocyte-like and inflammatory cells, and blood-filled spaces, mimicking vascular channels but lacking an endothelial lining. Immunohistochemical analysis (performed with the following monoclonal antibodies: smooth muscle actin, vimentin, desmin, CD-31, CD 34, CD-68) showed only vimentin positively on neoplastic cells. We discuss the differential diagnostic hypotheses and review the literature on this subject. PMID- 11428293 TI - [Herceptin: a new drug finding new life in the determination of c-erb-B2 in breast cancer]. PMID- 11428294 TI - [Wegener's granulomatosis: role of anatomopathology in early diagnosis, with particular reference to pulmonary and nasal sinus biopsy]. PMID- 11428295 TI - [Cancer as a genetic disease]. PMID- 11428296 TI - Cytometric investigations of bladder irrigation/washing and voided urine. AB - Several studies have shown that cytometry, including DNA analysis, gives valuable information on the grade and stage of bladder cancer when performed on cytological preparations obtained from urine. Although, cytometry should not be used as a screening tool, it has a role in the follow-up of patients with a previous history of superficial bladder cancer. In this group of patients, the combination of cytological examination with cytometric evaluation allows the detection of the majority of recurrent tumors. In patients treated with chemotherapy or immunotherapy, the presence or the appearance of an aneuploid cell population is a good indication of tumor recurrence or progression. A variety of wet laboratory immunoassays, on-slide immunoassays, in situ hybridization procedures and post-nucleic acid extraction molecular techniques have been designed to complement cytology and cytometry and to improve the overall sensitivity and specificity of the detection of recurrent urothelial neoplasia. PMID- 11428298 TI - [Science and man. Inauguration of the academic year 2000-2001 of the Bologna University]. PMID- 11428297 TI - [Sovereign illuminated, Napoleon, Pavia, morphology: the anatomic museum of Bologna]. PMID- 11428299 TI - [Silver-stained Nucleolar Organizer Regions (AgNOR)]. AB - The present paper discusses the relevance of silver-stained Nucleolar Organizer Regions (AgNOR) to tumour pathology. First, the structural and functional aspects of AgNORs and the biological meaning of their quantitative variations have been reviewed to clearly define which indications can be obtained by the use of AgNOR. In continuously proliferating cells, AgNOR indicates the extent of ribosomal biogenesis, which is strictly related to the rapidity of cell proliferation. Therefore, AgNOR can be considered to represent a marker of cell proliferation rate and, in tumour pathology, should be used only for prognostic purposes. The predictive value of AgNOR, the only parameter which indicates the cell growth rate in situ in routinely processed cyto-histological samples, is strengthened by the combination with markers of cell growth kinetics (e.g. Ki67/MIB1). PMID- 11428300 TI - ["If I were the department head in this case..."--interventional interview with patients and co-workers of a psychiatric department]. AB - OBJECTIVE: How can patients and staff participate in redesigning psychiatric procedures through survey research? METHODS: Using interviews with circular hypothetical questioning, 58 patients and 30 staff members of a psychiatric department of a general hospital were interviewed about their preferences for change and continuity in clinical practices, and results were feedbacked. RESULTS: Suggestions for change concerning doctors' rounds, the integration of music therapy, the number of individual sessions and the postdischarge care initiated real change. CONCLUSIONS: Surveys planned cooperatively with staff and without competitive benchmarking can be effective tools in psychiatric organization development. PMID- 11428301 TI - [Burnout in psychiatric physicians. Results of an empirical study]. AB - OBJECTIVE: The study aimed at assessing burnout in psychiatrists and psychiatric residents, all of them working in the Canton of Zurich, Switzerland, and at identifying some of its correlates. METHODS: Potentially burnout- related demographic, work, leisure activities, and personality (Munich Personality Test) variables were studied in three subgroups of 307 probands--psychiatric residents, psychiatrists working as staff members or in leading positions in psychiatric institutions, and psychiatrists working in private practice--the degree of burnout having been assessed with the Tedium Measure (TM). RESULTS: The mean TM score of all probands of 2.9 (SD = 0.7) was on a low level, high TM scores (> 3.5) were indicated by 57 (18%) participants. Significantly higher TM scores were indicated by psychiatric residents, who also scored higher with regard to neuroticism and lower with regard to frustration tolerance on MPT. CONCLUSIONS: Neuroticism alone explained a substantial proportion of the total TM variance in individual groups. Work-related variables turned out to be of a small importance only, whereas no influence could be demonstrated for different leisure activities. PMID- 11428302 TI - ["(...) sometimes I have doubts about myself, when I'm not doing well". Effect of an ambulatory psychoeducational group program from the subjective perspective of patients with schizophrenic illnesses]. AB - PURPOSE: Aim of the study is an analysis of the patients' evaluation of the impact of a psychoedukative group program for patients with schizophrenia in outpatient treatment, which was part of the project "Integrated Treatment of Schizophrenia". METHOD: Problem-focussed interviews were carried out with 30 randomly selected participants of the psychoeducative group programme. Interviews were fully transcribed and analysed by means of qualitative content analysis. RESULTS: With the exception of two persons all patients made a very positive assessment of their participation at the psychoeducative programme. The quality of the information received and the possibilities of exchanging experiences with other patients during the group sessions were mentioned by most of the participants as reasons for this positive assessment. Although only few patients reported on changes of their illness behaviour due as a result of psychoeducation, most of them underlined that that the increase of their illness related wisdom as well as the exchange of illness experiences with other patients helped them to cope somewhat better with their illness. CONCLUSION: Psychoeducative group programmes contribute to the improvement of patients quality of life even if they have no measurable impact on illness behaviour or compliance. Hence, such group programmes should be integrated into outpatient services even if there is no evidence of a direct impact on relapse rates or cost savings. PMID- 11428303 TI - [Etiology of schizophrenia from the viewpoint of the patient]. AB - OBJECTIVE: According to a quantitative study on causal beliefs patients with functional psychoses favour psychosocial over biological explanations. They also tend to endorse a multicausal concept. The aim of this study is to try to replicate these findings using semiqualitative methods. METHODS: Problem-centered interviews were conducted with 78 schizophrenic patients consecutively discharged from inpatient or dayhospital treatment. RESULTS: As in the previous study, patients provided more frequently psychosocial explanations. Biological causes were relatively rarely mentioned--mostly in response to suggestions of the interviewer. In addition, they were rather brief and not very elaborated. By contrast to the previous study, hardly any indications for a multicausal concept could be observed. DISCUSSION: Patients' beliefs are contrasted with the results of psychiatric research on the causes of schizophrenia. PMID- 11428304 TI - [Memory performance and instruction effects in schizophrenia. A comparative study of chronic schizophrenic patients and healthy probands]. AB - 34 chronically schizophrenic persons were matched with 34 healthy control participants according to age, level of education, and sex and received memory tests (short-term, priming, working, prospective, episodic, source and semantic long-term memory) and tests of attention and intelligence. Patients performed worse on all tests. The patients' memory deficits compared with controls are nearly one standard deviation greater than the differences in attention and intelligence. The deficits are smaller for priming and semantic long-term memory than for short-term, working, prospective, episodic, and source memory. After one short instruction all patients and controls received a repeat administration of the tests. Patients showed a lower, but also significant improvement for short term, working, and prospective memory. We conclude that schizophrenia patients might suffer from specific memory deficits (specific vulnerability of strategic and thus of interference- or context-sensitive memory tasks) not attributable to more general psychopathologic symptoms. PMID- 11428305 TI - [Phenomenology of heautoscopy. Case report of atypical mirror hallucination]. AB - Heautoscopy is a well-known phenomenon in clinical psychopathology. Strictly speaking it is not a syndrome but a symptom and can be associated with different kinds of mental illness. Our case report deals with an atypical form of heautoscopy. We try to elucidate the essential qualities of our patient's disturbing mental experiences. In addition, we present a psychodynamic approach which seeks to explain the patient's experiences in terms of projection and protective functions. PMID- 11428306 TI - [Psychogenic seizures: domain for a hoca?]. AB - We report a case of an 18 year old female Turkish patient, who was suffering from psychogenic seizures. The family did not accept the disease, prohibited treatment by a medical doctor, preferred to bring her to a Hoca. She was maltreated by her father und sexually abused at the age of eight years by her brother. In this article we describe the problems of a psychiatric ill female patient within a family of Turkish origin. PMID- 11428307 TI - [Neuropsychological follow-up of attention deficit/hyperactive disorder in adulthood before and after treatment with methylphenidate]. AB - Attention-deficit/hyperactivity disorder (ADHD) in adulthood is frequently overlooked. Motor restlessness, impulsivity, low frustration tolerance, and conduct disorders are diagnostically ambiguous, but are predominantly ascribed to affective disorders, anxiety disorders, and personality disorders. Furthermore, substance-induced addiction may develop consecutively to untreated ADHD. We report on a case of primary diagnosis of ADHD in adulthood with secondary cocaine dependency, who was neuropsychologically assessed before and following treatment with methylphenidate. Frequency of errors and reaction time in performance on attention tests, concentration tasks, and multiple stimuli-/reaction tasks improved under treatment with methylphenidate. ADHD may be treated efficiently even after primary diagnosis in adulthood. Differential diagnosis of ADHD should be considered in disorders associated with motor hyperactivity, impulsivity, stress intolerance and conduct disorders. PMID- 11428308 TI - [Decentralized geriatric psychiatry. Experiences and evaluation of developing a memory consultation clinic at a regional hospital]. AB - PURPOSE: At the General Hospital in Thun (Switzerland) a memory clinic was established in 1999. METHOD: The first 55 cases (30 females and 25 males) were statistically analysed as a part of internal quality management. The patients were examined using a usual assessment including physical and psychiatric examination, blood and neuropsychological testing (Alzheimer's Disease Assessment Scale). RESULTS: 14% of the examined patients did show kind of cognitive deficit. Men yielded significantly better results in neuropsychological testing than women. CONCLUSIONS: The general acceptance of the new memory clinic in Thun is due to the growing demand for such institutions because of the demographic development. Men's better results in neurological testing might be a sign of different reaction-patterns of their families, that make family practitioners send them at an earlier stage of dementia. PMID- 11428309 TI - [Polyneuropathy and myelopathy in N2O abuse within the scope of multiple drug abuse]. PMID- 11428310 TI - The design of a corporate identity for a department of medical illustration: a case study. AB - This paper outlines the author's attempt to design and introduce a corporate identity into the Department of Medical Illustration at the South Buckinghamshire NHS Trust. It is intended to furnish the reader with an insight into the process of designing a corporate identity and to relate one department's experience. This may be useful for those who wish to develop a corporate identity of their own or contribute, as a department, towards an identity for their own Trust or other institution. A major change in government policy about the identity of NHS Trusts has meant that use of the department's new logo has had to be abandoned in favour of the new NHS corporate identity. PMID- 11428311 TI - Research for medical illustrators: searching for references. AB - The proliferation of online databases means that medical illustrators have ready access to a wide range of resources for research. This paper looks at approaches to finding references from a systematic keyword search, published bibliographies and journals of abstracts, to locally and nationally available online databases, e-mail alerting services and full-text journals. The Internet will often provide an abstract (similar to this), however, it may still be necessary to visit a library, archive or other source to find a hard copy, particularly for older material. PMID- 11428312 TI - What's your style? PMID- 11428313 TI - Double-vision: stereo photography in ophthalmology. PMID- 11428314 TI - Adobe Photoshop and Illustrator techniques. PMID- 11428315 TI - Virtual reality applied to medicine, IMI Trent regional meeting, 25 October 2000. PMID- 11428316 TI - The Prince of Wales Hospital archive (1930-1942). PMID- 11428317 TI - Future direction of medical genetics. PMID- 11428318 TI - Double contrast barium enema and colorectal carcinoma: sensitivity and potential role in screening. AB - To establish the sensitivity of double contrast barium enema (DCBE) for detection of colorectal carcinoma in a tertiary referral centre and consider its possible role as a suitable imaging method in screening for this disease. A total of 160 patients with a histopathologically proven diagnosis of colorectal carcinoma over a two year period were reviewed. Subsequently 112 of the 160 patients were identified as having undergone DCBE, the results of which were analysed to determine its sensitivity for detecting colorectal carcinoma. Colorectal carcinoma was missed in 4 of the 112 barium enemas performed. This corresponds to a sensitivity of 96.5% with a false negative rate of 3.5%. The Dukes Classification in these 4 cases showed that Dukes stage B, C and D were missed, with tumours located in the right and the sigmoid colon. The mean delay to operation in these four cases was 6 weeks. Our study correlates with previous studies showing a false negative rate for DCBE of 3.5%. Colonoscopy also fails to detect small numbers of tumours with false negative rates reported as high as 10%. We suggest that double contrast barium enema should be effective as a screening method in any future colorectal cancer screening program. PMID- 11428319 TI - Ionising radiation (medical exposure) regulations (Northern Ireland) 2000 and their implications for Accident and Emergency (A&E) doctors in training. AB - With the introduction of the Ionising Radiation (Medical Exposure) Regulations (Northern Ireland) 2000 (IRMER) the medical practitioner faces greater accountability when requesting radiological investigations. The referrer (usually a doctor or dentist) must supply sufficient medical data to justify radiation exposure to a patient. These regulations can lead to criminal prosecution if breached. Our objectives were to identify the level of unjustified requests for plain abdominal radiography among A&E doctors and whether there is a statistically significant difference in the justification of request between doctors of differing experience. We reviewed and prepared statistical analysis of 100 A&E request forms for plain abdominal radiography. Royal College of Radiologist Guidelines were used as a "Gold standard" for justification of the investigation. A&E doctors of less than six months experience are at greater risk of breaching these regulations when requesting plain abdominal films, when compared to more experienced doctors. This is a serious issue which should be addressed at undergraduate and pre-registration level in addition to ongoing audit. PMID- 11428320 TI - Absence of the palmaris longus muscle: a population study. AB - We examined 300 Caucasian subjects (150 males, 150 females) aged 18-40 years to assess the incidence of palmaris longus absence. The presence or absence of palmaris longus was assessed by clinical inspection. Forty-nine subjects had unilateral absence of palmaris longus (16%). The tendon was absent bilaterally in 26 subjects (9%). Unilateral and bilateral absence was more common in males, however this was not statistically significant (p = 0.25 and 0.56 respectively). In those subjects with unilateral absence, the right side was found to be more commonly affected however no statistical significance was evident (p = 0.25). PMID- 11428321 TI - Abnormal uterine bleeding: an evaluation endometrial biopsy, vaginal ultrasound and outpatient hysteroscopy. AB - Abnormal uterine bleeding is a common gynaecological symptom. Whilst most patients have benign disease, thorough investigation is necessary, particularly in the peri- and post-menopausal woman. Hysteroscopy with directed biopsy of suspicious lesions is the gold standard investigation but it is invasive and is not offered in all units as an outpatient procedure. Ultrasound and outpatient biopsy techniques may allow patients to be triaged to select those who require formal evaluation by hysteroscopy. We assessed the records of 100 consecutive referrals to the outpatient hysteroscopy clinic at the Royal Maternity Hospital, Belfast. In order to develop a nomogram for the investigation of women with abnormal bleeding patterns, we compared ultrasound/biopsy diagnoses with hysteroscopy/biopsy diagnosis. Pre- and peri-menopausal patients, and post menopausal patients taking hormone replacement therapy should be investigated further if the endometrial thickness is more than 10 mm, if endometrial biopsy is abnormal, or if symptoms are recurrent. Similarly, post-menopausal patients not taking hormone replacement therapy with an endometrial thickness of 5 mm or more should be referred for hysteroscopy. PMID- 11428322 TI - Idiopathic intracranial hypertension; incidence, presenting features and outcome in Northern Ireland (1991-1995). AB - OBJECTIVES: To determine the age and sex specific incidence rates, presenting features, and visual outcome of idiopathic intracranial hypertension in Northern Ireland. METHODS: A case-note review of all patients with idiopathic intracranial hypertension, diagnosed at the Royal Victoria Hospital, Belfast between 1991 and 1995. RESULTS: Forty-two patients were identified corresponding to an average annual incidence rate per 100,000 persons of 0.5 for the total and 0.9 for the female population. The commonest presenting symptoms were headache (84%), transient visual obscurations (61%) and sustained visual loss (34%). Impaired Snellen visual acuity and visual field loss were documented in 21% and 62% of patients respectively at presentation, and in 24% and 39% at last follow-up. One patient suffered deterioration in visual functioning sufficient to interfere with normal daily activities. CONCLUSIONS: The age and sex specific incidence rates of IIH in Northern Ireland are lower than have been reported in previous population based series. Disabling visual loss occurs in a small number of patients despite all interventions. PMID- 11428323 TI - In search of optimism. PMID- 11428324 TI - Familial craniosynostosis due to Pro250Arg mutation in the fibroblast growth factor receptor 3 gene. PMID- 11428325 TI - The history of the teaching of the specialty of general practice in Northern Ireland. Presidential address to the Ulster Medical Society. PMID- 11428326 TI - Endovascular repair for acute symptomatic and ruptured abdominal aortic aneurysms. PMID- 11428327 TI - Gastric variceal haemorrhage successfully managed by splenectomy--a case report and literature review. PMID- 11428328 TI - Cronkhite-Canada syndrome. PMID- 11428329 TI - Cystic dilatation of the rete testis. PMID- 11428330 TI - A fatal case of carcinoma arising from a pilonidal sinus tract. AB - We report a male patient with carcinoma arising on the basis of neglected sacrococcygeal pilonidal sinus disease. Following initial operation, performed without suspicion of malignancy, histology demonstrated cellular atypia and an increased mitotic rate. A second, wider tissue excision was recommended but the patient declined further surgery. Two years later, he presented with fungating carcinoma involving the rectum but again declined surgery. This rare case demonstrates that the presence of carcinoma should be suspected in long-standing, although innocent-looking, pilonidal sinus disease. In the circumstance of uncertain histologic diagnosis, more generous surgical sampling is required. Every effort must be made to overcome patient's reluctance to accept a second, possibly life-saving procedure. PMID- 11428331 TI - Dysphagia due to an aberrant left subclavian artery in a right-sided aortic arch. PMID- 11428332 TI - Review of referrals for the FISH detection of Williams syndrome highlights the importance of testing in supravalvular aortic stenosis/pulmonary stenosis. PMID- 11428333 TI - Perch (Perca fluviatilis L.) parasites reflect ecosystem conditions: a comparison of a natural lake and two acidic reservoirs in Finland. AB - Parasite communities of perch were studied in a natural lake (Vetamajarvi) and two reservoirs (the Kyrkosjarvi and Liikapuro reservoirs) located in Western Finland. All water bodies studied are small, shallow and humic. However, the reservoirs are more acidic than the lake (pH 5.9 and 5.3 vs. 6.4). Altogether, 18 parasite species were found, but the component communities were reduced in the harsh conditions of the reservoirs (12 and six species) as compared with the lake (17 species). In addition, the mean number of metazoan parasite specimens per fish was markedly lower in the reservoirs (64.3 and 14.3 specimens) than in the lake (116.1 specimens). Our prediction on the depauperation of parasite communities in reservoirs as compared with the lake was confirmed. Many ectoparasite groups were rare and most of the digenean parasites using molluscs as their intermediate host were absent from the reservoirs due to the inability of molluscs to tolerate the acidic water. The only digenean parasite found in the less acidic reservoir was Bunodera luciopercae. There were also species that preferred reservoirs as was expected. The protozoan ciliate Capriniana piscium benefiting acidic water and Acanthocephalus lucii, due to the abundance of its intermediate host, were most prevalent in the reservoir conditions. PMID- 11428334 TI - Parasites of recruiting coral reef fish larvae in New Caledonia. AB - Recruiting coral reef fish larvae from 38 species and 19 families from New Caledonia were examined for parasites. We found 13 parasite species (Platyhelminthes: Monogenea, Cestoda and Trematoda) but no acanthocephalan, crustacean or nematode parasites. Over 23% of individual fish were infected. Didymozoid metacercariae were the most abundant parasites. We conclude that most of the parasites are pelagic species that become 'lost' once the fish larvae have recruited to the reef. Larval coral reef fish probably contribute little to the dispersal of the parasites of the adult fish so that parasite dispersal is more difficult than that of the fish themselves. PMID- 11428335 TI - Differential influence of Pomphorhynchus laevis (Acanthocephala) on the behaviour of native and invader gammarid species. AB - Although various species of acanthocephalan parasites can increase the vulnerability of their amphipod intermediate hosts to predation, particularly by altering their photophobic behaviour, their influence on the structure of amphipod communities and the success of invader species has so far received little attention. We compared the prevalence and behavioural influence of a fish acanthocephalan parasite, Pomphorhynchus laevis, in two species of amphipods, Gammarus pulex and Gammarus roeseli in sympatry in the river Ouche (Burgundy, eastern France). There, G. pulex is a resident species, whereas G. roeseli is a recent coloniser. Both uninfected G. pulex and G. roeseli were strongly photophobic. although less so in the invading species. However, there was no significant difference in reaction to light between infected and uninfected G. roeseli, whereas infected G. pulex were strongly photophilic. We discuss our results in relation to the parasite's ability to manipulate invading host species, the possibility that resistant individuals have been selected during the invasion process, and the role that acanthocephalan parasites can play in shaping the structure of amphipod communities. PMID- 11428336 TI - Regional genetic variation in the major sperm protein genes of Onchocerca volvulus and Mansonella ozzardi (Nematoda: Filarioidea). AB - Onchocerca volvulus and Mansonella ozzardi are two human filarial parasites present in South and Central America. In the Brazilian Amazonia they are found in sympatry, and the lack of clear morphological diagnostic characters in the microfilariae hinders their identification. The major sperm protein (MSP) gene of both species has been sequenced and characterised to determine its potential as a molecular diagnostic character. The length of the MSP gene is different in each species, and this could be used to detect and differentiate them by running the polymerase chain reaction (PCR) product in an agarose gel. Two major gene groups were identified in O. volvulus with a genetic distance of 6% between them. In M. ozzardi only one major group of genes was observed. The high similarity between the protein amino acid sequence of both filarial species confirms that the MSP has been highly conserved through nematode evolution. PMID- 11428337 TI - The production of a 70 kDa heat shock protein by Toxoplasma gondii RH strain in immunocompromised mice. AB - A 70 kDa heat shock protein (HSP70) has been reported previously to be strongly expressed in virulent Toxoplasma gondii strains taken from immunocompetent mice but it is poorly expressed by virulent parasites in mice immunocompromised by treatment with cortisone acetate or by virulent parasites cultured in vitro. Immune factors such as interferon-gamma, tumour necrosis factor and reactive nitrogen intermediates derived from nitric oxide are known to be important inducers of HSP70 production and are also known to be produced during the immune response to acute T. gondii infection. The ability of these immune factors to induce T. gondii HSP70 production was tested by analysing HSP70 production in tachyzoites of the virulent RH strain of T. gondii recovered from mice deficient in: (1) T cells (nude mice); (2) T and B cells (SCID mice); (3) interferon-gamma receptors (interferon-gamma receptor knockout mice); and (4) tumour necrosis factor receptors (tumour necrosis factor receptor knockout mice). Parasites from nude and SCID mice produced as much HSP70 as immunocompetent mice. Likewise, T. gondii tachyzoites from mice lacking receptors for interferon-gamma or tumour necrosis factor produced HSP70 in quantities similar to wild-type mice. The ability to produce reactive nitrogen intermediates in response to T. gondii infection, as detected by elevated levels of nitrate and nitrite in sera, was normal in tumour necrosis factor receptor knockout mice but was completely lacking in interferon-gamma receptor knockout mice, indicating that reactive nitrogen intermediates are also not responsible for induction of parasite HSP70. Thus, immune factors that induce HSP70 production in mammalian cells do not appear to play primary roles in inducing HSP70 production by T. gondii. PMID- 11428338 TI - Trypanosoma cruzi: skin-penetration kinetics of vector-derived metacyclic trypomastigotes. AB - In order to investigate the natural route of infection of nude and normal BALB/c mice with Trypanosoma cruzi via the skin, a drop of vector faeces/urine containing metacyclic trypomastigotes was placed onto the puncture site of a bite from Triatoma infestans. The periods of exposure, i.e. until removal of flagellates from the skin, and the time elapsed until surgical removal of the skin around the puncture were varied. After 15 min of exposure, T. cruzi developed in all nude mice without surgery, and in four of 10 mice if the puncture region of the skin was removed directly after exposure. In a shaved puncture region, 5 min of exposure were sufficient to infect all normal BALB/c mice without surgery and one of four mice with direct removal of the puncture region. Longer periods of exposure or time until removal of the skin only sometimes resulted in higher infection rates. Prepatent periods and the development of parasitaemia varied irrespective of the period of exposure or the period until skin removal at the puncture site. The importance of these findings is that they clearly prove that T. cruzi can rapidly invade the host via the puncture site of the bite of the vector and that at least some parasites are immediately transported away from this site. PMID- 11428339 TI - The role of oxygen availability in the embryonation of Heterakis gallinarum eggs. AB - The importance of oxygen availability in the embryonation of the infective egg stages of the gastrointestinal nematode parasite Heterakis gallinarum was studied in the laboratory. Unembryonated H. gallinarum eggs were kept under either aerobic conditions by gassing with oxygen, or anaerobic conditions by gassing with the inert gas nitrogen, under a range of constant temperatures. Oxygenated eggs embryonated at a rate influenced by temperature. Conversely, eggs treated with nitrogen showed no embryonation although when these eggs were transferred from nitrogen to oxygen gas after 60 days of treatment, embryonation occurred. This demonstrated that oxygen is an essential requirement for H. gallinarum egg development, although undeveloped eggs remain viable, even after 60 days in low oxygen conditions. The effects of climate on the biology of free-living stages studied under constant laboratory conditions cannot be applied directly to the field where climatic factors exhibit daily cycles. The effect of fluctuating temperature on development was investigated by including an additional temperature group in which H. gallinarum eggs were kept under daily temperature cycles between 12 and 22 degrees C. Cycles caused eggs to develop significantly earlier than those in the constant mean cycle temperature, 17 degrees C, but significantly slower than those in constant 22 degrees C suggesting that daily temperature cycles had an accelerating effect on H. gallinarum egg embryonation but did not accelerate to the higher temperature. These results suggest that daily fluctuations in temperature influence development of the free-living stages and so development cannot be accurately predicted on the basis of constant temperature culture. PMID- 11428340 TI - Serodiagnosis of amoebiasis using a recombinant protein fragment of the 29 kDa surface antigen of Entamoeba histolytica. AB - To develop an improved serodiagnostic test for amoebiasis, we performed a detailed analysis of the immunodominant epitopes of the 29 kDa surface antigen and evaluated its sensitivity and specificity. Enzyme-linked immunosorbent assay (ELISA) based on the fragment containing the immunodominant epitope was evaluated further and compared with full-length recombinant 29 kDa protein. Specificity and sensitivity of the two ELISAs were assessed using 55 human sera of parasitic protozoa infection cases (25 amoebiasis, 20 giardiasis and 10 toxoplasmosis sera) and 10 healthy control sera. The immunodominant epitope of the 29 kDa antigen is localised only in the N-terminus 14-54 amino acid residues. The sensitivities of the two ELISAs were very high, 92 and 96%, respectively. The specificity of the fragment was 100%, whereas the specificity of the full-length 29 kDa protein was 86.6%. These results indicate that the fragment containing the immunodominant epitope of the 29 kDa protein can be used to accurately serodiagnose amoebiasis without cross-reactivity from other parasites. PMID- 11428341 TI - Mapping and expression of microneme genes in Eimeria tenella. AB - Microneme organelles are located at the apical tip of invading stages of all apicomplexan parasites and they contain proteins that are critical for parasite adhesion to host cells. In this paper, we have utilised the process of oocyst sporulation in Eimeria tenella to investigate the timing of expression of components of the microneme organelle, at both mRNA and protein levels. Two time course studies showed that there is a high level of synchrony in the sporulation process, especially during the time period when sporozoites are formed. Western blotting showed that the expression of five microneme proteins (EtMIC1-5) is differentially regulated and highly co-ordinated during sporulation with the proteins being detected only towards the end of the process, as the sporozoites matured within the sporocysts. In contrast, mRNA for all five of these microneme proteins was detected some 10-12 h earlier in sporulation than when the corresponding proteins were seen. Overall these data suggest that the expression of proteins destined for the microneme is regulated both at the transcriptional and translational level. The single copy genes encoding EtMIC1-5 are not clustered on the genome, but are found on four different chromosomes. PMID- 11428342 TI - There are at least three genetically distinct small piroplasms from dogs. AB - The 18S nuclear subunit ribosomal RNA (18S rRNA) gene of small piroplasms isolated from dogs from Okinawa (Japan), Oklahoma, North Carolina, Indiana, Missouri, and Alabama, was isolated and sequenced. Phylogenetic analysis of these sequences and comparisons with sequences from other Babesia, Cytauxzoon, and Theileria species revealed that all canine small babesial isolates, with the exception of isolates from California and Spain, were placed in a group containing the Babesia spp. sensu stricto. Within the Babesia spp. sensu stricto, there was support for separating the small canine piroplasms from the large canine piroplasm, Babesia canis. The isolate from California was in a distinct phylogenetic clade, closely related to babesial isolates from wildlife and humans from the Western US. The canine isolate from Spain was closely related to Babesia microti. These results suggest that there are multiple small piroplasm species in dogs. The isolates from the Midwestern and Eastern US and the one from Japan probably represent a single species with wide geographic distribution. PMID- 11428343 TI - Gene transfer of endothelial nitric oxide synthase to the penis augments erectile responses in the aged rat. PMID- 11428344 TI - Erectile dysfunction in cyclic GMP-dependent kinase I-deficient mice. PMID- 11428345 TI - [Occupational exposures to blood-borne pathogens in health care workers]. AB - OBJECTIVE: To determine the rate of occupational exposures to blood-borne pathogens in different occupations of health care workers. To analyze the characteristics and outcome of the occupational exposure. MATERIAL AND METHODS: We have evaluate occupational exposures to blood-borne pathogens reported by health care workers during 1996-1999. The following data were collected: characteristics of the workers, type of occupational exposure, immunity status of the exposed worker, infectivity of the source patient and follow up serologic testing of the worker. RESULTS: A total of 407 occupational exposures were reported. The highest rate of occupational exposure was found among nurses (61.6%). Needlestick accident was the most often occupational exposure reported (84.5%). Mucosal exposures with accidental splashes were reported in 15.2% of cases. In 14.5% of these accidents workers were at risk for occupational transmission of blood-borne pathogens. Among the different occupations of health care workers, the rate of exposures with a source infected patient was higher in medical staffs (28.3%) than nurses (13.9%) The rate of exposures with a source infected patient was higher in accidental splashes than in percutaneous exposures (33.8% vs 13.3%), besides in none of the accidental splashes, employees had used appropriate barrier precautions. There were no cases of transmission of occupational blood-borne infections. CONCLUSIONS: Although nurses are the health care workers with highest rates of occupational exposures, medical staffs are the most often occupationally exposed to a source infected patient. Universal barrier precautions are no appropriately used in most of the occupational accidents, specially in those involving mucosal exposures. PMID- 11428346 TI - Genetic polymorphisms of the dopamine D2 and D3 receptor and neuroleptic drug effects in schizophrenic patients. PMID- 11428347 TI - Clinical and biochemical correlates of "high-dose" clozapine therapy for treatment-refractory schizophrenia. PMID- 11428348 TI - The relationship between mental status examination scores and cerebral blood flow in geriatric schizophrenia. PMID- 11428349 TI - Financial thoughts for the new dentist. PMID- 11428350 TI - Getting started in dentistry: reflections at six months. PMID- 11428352 TI - A dental analogy to financial planning. PMID- 11428351 TI - Membership directory of the Colorado Dental Association. Directory 2000/2001. PMID- 11428353 TI - Hire qualified assistants, know delegable tasks. PMID- 11428354 TI - Guidelines for reporting clinical features in cases with MECP2 mutations, by Allison Kerr and an international group. PMID- 11428355 TI - Visual vignette. A case presentation: eruptive xanthoma. PMID- 11428356 TI - AACE medical guidelines for clinical practice for the diagnosis and treatment of dyslipidemia and prevention of atherogenesis. PMID- 11428357 TI - Visual vignette. A case presentation: giant mucocele. PMID- 11428358 TI - Albright's hereditary osteodystrophy: historical credit. PMID- 11428359 TI - The American Association of Clinical Endocrinologists Medical Guidelines for the Management of Diabetes Mellitus: the AACE system of intensive diabetes self management--2000 update. PMID- 11428360 TI - An alternative estimate of the disappearance rate of ventricular fibrillation in our-of-hospital cardiac arrest in Sweden. PMID- 11428361 TI - Antioxidant strategy for cardiovascular disease. PMID- 11428362 TI - Antioxidant strategy for cardiovascular diseases. PMID- 11428363 TI - Antioxidant strategy for cardiovascular diseases. PMID- 11428364 TI - Cerebral venous thrombosis. PMID- 11428365 TI - Dopamine dose and renal damage. PMID- 11428366 TI - Direct observation of tuberculosis treatment. PMID- 11428367 TI - Propofol use in head-injury patients. PMID- 11428368 TI - Congenital absence of inferior vena cava. PMID- 11428369 TI - What is happening with hand transplants. PMID- 11428370 TI - Familial hypercholesterolaemia. PMID- 11428371 TI - Japan's new long-term care insurance. PMID- 11428372 TI - Dutch-Romanian connection on family medicine. PMID- 11428373 TI - Health warning to visitors to the Indian sub continent. PMID- 11428374 TI - Human rabies and bat bites. PMID- 11428375 TI - Restaurant employment boom in New York City. PMID- 11428376 TI - An evaluation of the impact of the introduction of "non-smoking" areas on trade and customer satisfaction in 11 public houses in Staffordshire. PMID- 11428377 TI - [A-56-year-old woman with parkinsonism, whose mother had Parkinson's disease]. AB - We report a 56-year-old woman with progressive gait disturbance. Her mother had Parkinson's disease with onset at age 70. She died at age 74 and the post-mortem examination confirmed the diagnosis of Lewy body positive Parkinson's disease. The patient was well until the age of 50(1995) when she noted an onset of resting tremor and difficulty of gait. She also developed delusional ideation and was admitted to a psychiatric service of another hospital, where a major tranquilizer was given. The delusion disappeared but she developed marked rigidity. The major tranquilizer was discontinued and an anticholinergic and amantadine HCl were given. She showed marked improvement to Hoehn and Yahr stage II and was discharged. In 1995, when she was 52 years of the age, she developed delusion again and a major tranquilizer was given. She developed marked parkinsonism again and became Hoehn and Yahr stage V. The major tranquilizer was discontinued and she was treated with levodopa/carbidopa, trihexyphenidyl, bromocriptine, and dops. She improved remarkably to stage II. She was admitted to our service on October 8, 1996 for drug adjustment. She was alert and not demented. She was anxious but delusion or hallucination was noted. Higher cerebral functions were intact. Cranial nerve functions were also intact except for masked face and small voice. Her posture was stooped and steps were small. She showed retropulsion and moderate bradykinesia. Resting tremor was noted in her left hand. Rigidity was noted in both legs. No cerebellar ataxia or weakness was noted. Deep tendon reflexes were within normal range and sensation was intact. Her cranial MRI revealed some atrophic changes in the putamen, in which a T 2-high signal linear lesion was seen along the lateral border of the putamen bilaterally. In addition, posterior part of the putamen showed T 2-low signal intensity change. She was treated with 1.6 mg of talipexole, 6 mg of trihexyphenidyl, and 100 mg of L-dops. She was in stage III of Hoehn and Yahr. She developed neurogenic bladder with a large amount of residual urine for which she required catheterization. She was transferred to another hospital. Despite drug adjustment, she lost response to levodopa and her parkinsonism deteriorated gradually. She also developed syncope orthostatic hypotension. In April of 1998, she developed intracerebral hemorrhage and was admitted again on April 19, 1998. She was unable to stand and showed marked akinesia and rigidity. She was in stage V of Hoehn and Yahr. Her cranial CT scan revealed bilateral high-density lesions in the posterior parietal lobes. She developed dysphagia for which she required gastrostomy. She was transferred to another hospital but her clinical condition deteriorated further. On December 22, 1999, she developed fever and dyspnea and was admitted to our service again. She developed cardial arrest at the emergency room from hypoxia. She was resuscitated; however, she was comatose with loss of brain stem reflexes. Later on she developed generalized myoclonus. She developed cardiac arrest and pronounced dead on December 28, 1999. The patient was discussed in a neurological CPC. The chief discussant arrived at the conclusion that the patient had striatonigral degeneration because of poor response to levodopa in the later course, autonomic failures, and MRI changes. Some other participants thought that the patient had a form of familial Parkinson's disease. Opinions were divided into these two possibilities. Post-mortem examination revealed that the substantia nigra showed intense neuronal loss and gliosis, however, no Lewy bodies were seen. In addition, intracytoplasmic inclusions were seen in oligodendrocytes. The putamen was markedly atrophic in its posterior part with marked gliosis and neuronal loss. The ventromedial part of the pontine nucleus also showed neuronal loss and intracytoplasmic glial inclusions. Pathologic diagnosis was multiple system atrophy. In the parietal lobe, an arteriovenous malformation with bleeding was noted. This is very unique case. Although her mother had Lewy body-positive Parkinson's disease, the patient had Lewy body negative multiple system atrophy with a-synuclein-positive glial inclusions. Whether this is just a coincidental occurrence or the presence of a genetic load for Parkinson's disease might triggered her multiple system atrophy is an interesting question to be answered in future. PMID- 11428378 TI - Management of major pancreatic duct injuries in children. AB - BACKGROUND: The operative versus nonoperative management of major pancreatic ductal injuries in children remains controversial. The computed tomographic (CT) scan may not be accurate for determination of location and type of injury. We report our experience with ductal injury including the recent use of acute endoscopic retrograde cholangiopancreatography (ERCP) for definitive imaging, and an endoscopically placed stent as definitive treatment. This has not been reported in children. METHODS: In review of 14,245 admissions to a regional pediatric trauma center over a 14-year period, 18 patients with major ductal injuries from blunt trauma were noted. Records were reviewed for mechanism of injury, method of diagnosis, management, and outcome. RESULTS: There were 10 girls and 8 boys, ranging in age from 2 months to 13 years. The most common mechanisms of injury were motor vehicle and bicycle crashes. Admission CT scan in 16 children was suggestive of injury in 11, and missed the injury in 5. Distal pancreatectomy was carried out in eight patients with distal duct injuries: one died of central nervous system injury. Nonoperative management in three proximal duct injuries suggested by initial CT scan and in three missed distal duct injuries resulted in pseudocyst formation in five survivors; one patient died of central nervous system injuries. Two children with minimal abdominal pain, normal initial serum amylase, and no initial imaging developed pseudocysts. Two of seven pseudocysts spontaneously resolved and five were treated by delayed cystogastrostomy. Two recent children with suggestive CT scans were definitively diagnosed by acute ERCP and treated by endoscopic stenting. Clinical and chemical improvement was rapid and complete and the stents were removed. Follow-up ERCP, CT scan, and serum amylase levels are normal 1 year after injury. CONCLUSION: Pancreatic ductal injuries are rare in pediatric blunt trauma. CT scanning is suggestive but not accurate for the diagnosis of type and location of injury. Acute ERCP is safe and accurate in children, and may allow for definitive treatment of ductal injury by stenting in selected patients. If stenting is not possible, or fails, distal injuries are best treated by distal pancreatectomy; proximal injuries may be managed nonoperatively, allowing for the formation and uneventful drainage of a pseudocyst. PMID- 11428379 TI - The New Injury Severity Score and the evaluation of pediatric trauma. AB - BACKGROUND: To compare the effectiveness of the Injury Severity Score (ISS) and New Injury Severity Score (NISS) in predicting mortality in pediatric trauma patients. METHODS: NISS, the sum of the squares of a patient's three highest Abbreviated Injury Scale scores (regardless of body region), were calculated for 9,151 patients treated at four regional pediatric trauma centers and compared with previously calculated ISS values. The power of the two scoring systems to predict mortality was gauged through comparison of misclassification rates, receiver operating characteristic curves, and Hosmer-Lemeshow goodness-of-fit statistics. RESULTS: Although there were significant differences in mean NISS and ISS values for each hospital, differences in the predictive abilities of the two scoring systems were insignificant, even when analysis was restricted to the subgroup of patients with severe or penetrating injuries. CONCLUSION: The significant differences in the predictive abilities of the ISS and NISS reported in studies of adult trauma patients were not seen in this review of pediatric trauma patients. PMID- 11428380 TI - Re.: Severe head injury caused by airbag displacement. PMID- 11428381 TI - Re.: Nitroprusside in resuscitation of major torso trauma. PMID- 11428382 TI - Re.: Nitroprusside in resuscitation of major torso trauma. PMID- 11428383 TI - Coronary-bronchial blood flow and airway dimensions in exercise-induced syndromes. AB - 1. We have an incomplete understanding of integrative cardiopulmonary control during exercise and particularly during the postexercise period, when symptoms and signs of myocardial ischaemia and exercise-induced asthma not present during exercise may appear. 2. The hypothesis is advanced that baroreflex de-resetting during exercise recovery is normally associated with (i) a dominant sympathetic vasoconstrictor effect in the coronary circulation, which, when associated with obstructive coronary disease, may initiate a potentially positive-feedback cardiocardiac sympathetic reflex (variable myocardial ischaemia with symptoms and signs); and (ii) a dominant parasympathetic bronchoconstrictor effect in the presence of bronchovascular dilatation, which, when associated with raised mediator release in the bronchial wall, reinforces the tendency for airway obstruction (variable dyspnoea results). 3. There is a need for new techniques to examine hypotheses concerning autonomic control, during and after exercise, of the coronary and bronchial circulations and the dimensions of airways. Accordingly, a new ultrasonic instrument has been designed named an 'Airways Internal Diameter Assessment (AIDA) Sonomicrometer'. It combines pulsed Doppler flowmetry with transit-time sonomicrometry of airway circumference and single crystal sonomicrometry of airway wall thickness. Initial evaluation suggests it is relatively easy to apply during thoracotomy in recovery animals. The component devices are linear and will measure target variables with excellent accuracy. 4. In anaesthetized sheep, intubated with controlled ventilation, intravenous isoproterenol causes large increases in bronchial blood flow, a fall in arterial pressure and a reduction in airway circumference. This may reflect the dominant action of reflex vagal activity over direct beta-adrenoceptor inhibition of bronchial smooth muscle, the reflex source being baroreflex secondary to the fall in arterial pressure. These findings provide insight into the integrative mechanisms underlying the paradoxical negative effects sometimes observed when beta-adrenoceptor agonists are used in asthma. PMID- 11428384 TI - Neural mechanisms in the cardiovascular responses to acute central hypovolaemia. AB - 1. The haemodynamic response to acute central hypovolaemia consists of two phases. During phase I, arterial pressure is well maintained in the face of falling cardiac output (CO) by baroreceptor-mediated reflex vasoconstriction and cardio-acceleration. Phase II commences once CO has fallen to a critical level of 50-60% of its resting value, equivalent to loss of approximately 30% of blood volume. 2. During phase II, sympathetic vasoconstrictor and cardiac drive fall abruptly and cardiac vagal drive increases. In humans, this response is invariably associated with fainting and has been termed vasovagal syncope. 3. In both experimental animals and in humans, the responses to acute central hypovolaemia are greatly affected by anaesthetic agents, in that the compensatory responses during phase I (e.g. halothane) or their failure during phase II (e.g. alfentanil) are blunted or abolished. 4. Therefore, our present knowledge of the neurochemical basis of the response to hypovolaemia depends chiefly on the results of experiments in conscious animals. Use of techniques for simulating haemorrhage has greatly enhanced this research effort, by allowing the effects of multiple treatments on the response to acute central hypovolaemia to be tested in the same animal. 5. The results of such experiments indicate that phase II of the response to hypovolaemia is triggered, at least in part, by a signal from cardiac vagal afferents. There is also strong evidence that phase II depends on brainstem delta-opioid receptor and nitrergic mechanisms and can potentially be modulated by circulating or neuronally released adrenocorticotropic hormone, brainstem serotonergic pathways operating through 5-HT1A receptors and opioids acting through mu- and kappa-opioid receptors in the brainstem. 6. Phase II also appears to require input from supramedullary brain centres. Future studies should determine how these neurotransmitter systems interact and their precise neuroanatomical arrangements. PMID- 11428385 TI - Statistics in physiology and pharmacology: a slow and erratic learning curve. AB - 1. Learning how to apply statistical analyses to the results of experimental or clinical studies may take a lifetime of trial (and sometimes error), as it has done in the author's case. There is no evidence that biomedical investigators of the present generation are on a steeper learning curve. Gross misunderstandings of the purpose and functions of statistical analysis are apparent in applications to research grant-giving bodies and ethics committees, in manuscripts submitted to journals and sometimes in published papers. 2. Although estimation of minimal group (sample) size for a given power is an essential step in planning clinical studies, it seems to be used rarely in laboratory experimental work. This is despite exhortations to restrict the number of animals used to a minimum. 3. Most investigators use hypothesis testing to analyse their results, but their understanding of the meaning of the resultant P-values is slight. 4. A flaw found almost universally in biomedical manuscripts is to make multiple inferences from the results of a single study. The goal of statistical analysis is to maintain the familywise type I error rate (risk of false-positive inference) at a predetermined level (usually 5%). But, when multiple inferences are made from the same experiment, the risk of false-positive error is inflated. There are two solutions to this problem: (i) use a multiple comparison procedure to control the familywise type I error rate; and (ii) test a single, global hypothesis. 5. Biomedical investigators have been quick to acquire computer statistics software and to use it to analyse their experiments. However, they have been slow to recognize the limitations of this software. These include: (i) inadequate documentation of routines, so that neither the user nor the reader of published papers can be sure how the tests have been executed; (ii) flawed algorithms for the execution of statistical procedures; and (iii) failure to recognize that the best software for their purposes is that which takes them just beyond their statistical horizons. 6. The obvious solution to these difficulties is to recruit a biomedical statistician into every research group, at a relatively trivial cost. However, properly qualified biostatisticians are in desperately short supply in Australia. It follows that research groups, national grant-giving agencies and academic institutions must make provision for the proper training and subsequent employment of biostatisticians. PMID- 11428386 TI - Maternal prepregnancy weight and congenital heart defects in offspring. AB - To determine the relation between having an infant with a major heart defect and a mother's prepregnancy weight, we compared 1,049 Atlanta-area women who gave birth to liveborn or stillborn infants, each with a major heart defect, with 3,029 Atlanta-area women who gave birth to infants without birth defects. The infants of control women were randomly selected from birth certificates and were frequency-matched to the case group by race, birth hospital, and birth period from 1968 through 1980. After excluding diabetic mothers and adjusting for potential confounders, compared with average-weight women (body mass index 19.9- 22.7), we found that underweight women (body mass index <16.5) were less likely to have a child with a major isolated heart defect [odds ratio (OR) = 0.64; 95% confidence interval (CI) = 0.43--0.97], whereas the OR was elevated among overweight or obese women (body mass index >26) (OR = 1.36; 95% CI = 0.95--1.93). Using average-weight women who did not take periconceptional multivitamins as the reference group, periconceptional multivitamin use was associated with a reduced OR for isolated heart defects among average-weight women (OR = 0.61, 95% CI = 0.36--0.99) and underweight women but not among overweight or obese women (OR = 1.69, 95% CI = 0.69--3.84). PMID- 11428387 TI - Maternal caffeine intake and intrauterine growth retardation. AB - This study estimates the effect of maternal caffeine consumption throughout pregnancy on fetal growth. We studied 2,714 women who delivered a liveborn infant between 1988 and 1991. Detailed information regarding coffee, tea, and soda drinking during the first and third trimesters of pregnancy was obtained. Average caffeine intake during month 1 of pregnancy was higher than for month 7 (72.4 vs 54.0 mg per day). Consumption of >300 mg caffeine per day during month 1 (adjusted odds ratio = 0.91; 95% confidence interval = 0.44--1.90) and during month 7 (adjusted odds ratio = 1.00; 95% confidence interval = 0.37--2.70) was not associated with intrauterine growth retardation. There was little evidence for any effect modification due to cigarette smoking on the caffeine associations. This study provides evidence that antenatal caffeine consumption has no adverse effect on fetal growth. PMID- 11428388 TI - Surgical treatment of atrial fibrillation using radiofrequency energy. AB - BACKGROUND: The Maze III procedure for atrial fibrillation (AF) is effective but has not been used widely due to its complexity, bleeding risk, and added operative time. Surgical radiofrequency ablation may simplify the procedure and make intraoperative correction of AF more accessible and widely performed. METHODS: Endocardial pulmonary venous isolation was performed on 48 patients with AF undergoing concurrent operation using temperature-controlled radiofrequency energy delivered through a hand-held flexible probe. Additional right-sided lesions were made at the surgeon's discretion. RESULTS: Forty-two patients were appropriate for analysis (6 died). These patients had an AF duration of 4.8 +/- 6.4 years. At a mean follow-up of 138 +/- 96 days, 34 patients were in sinus rhythm. We were unable to demonstrate a difference in outcome based on AF duration, left atrial size, or addition of right-sided lesions. CONCLUSIONS: Radiofrequency atrial ablation was effective in 81% of patients with AF at restoring sinus rhythm at an average follow-up of 4 months. This procedure is simple to perform and should broaden the number of patients that receive an AF treatment procedure during concurrent cardiac operation. PMID- 11428389 TI - A BRCA1 variant, IVS23+1G-->A, causes abnormal RNA splicing by deleting exon 23. PMID- 11428390 TI - Acquired channelopathies in nerve injury and MS. AB - Although neurophysiologic doctrine has traditionally referred to "the" voltage gated sodium channel, it is now clear that there are at least nine genes that encode molecularly and physiologically distinct sodium channels. Mutations of sodium channel genes provide a basis for genetic channelopathies. Dysregulated expression of sodium channels due to alterations in activity of nonmutated channel genes, on the other hand, can produce acquired channelopathies. Two examples of acquired channelopathies are discussed in this article. Recent research has established that peripheral nerve injury can provoke an acquired channelopathy in spinal sensory neurons; axonal transection triggers the turning off of some previously active sodium channel genes and the turning-on of at least one previously silent sodium channel gene, a set of molecular changes that can result in hyperexcitability of these cells. Emerging evidence also suggests that an acquired channelopathy, characterized by abnormal expression of sensory neuron specific sodium channels that can alter impulse trafficking within Purkinje cells, may contribute to the pathophysiology of MS. Subtype-specific drugs that selectively modulate various types of channels probably will soon be developed. The acquired channelopathies associated with nerve injury and MS may thus represent prototype disorders that present therapeutic opportunities. PMID- 11428391 TI - Canine distemper in terrestrial carnivores: a review. AB - Canine distemper virus is a member of the genus Morbillivirus in the family Paramyxoviridae. Canine distemper has been recorded in domestic dogs for centuries. It is now recognized as a worldwide problem of carnivores and has the second highest fatality rate of any infectious disease, after rabies, in domestic dogs. The importance of this disease in nondomestic animals has become evident with vaccine-induced infections in a variety of species and large-scale epidemics in captive and free-ranging felids. To date, canine distemper has been reported in all families of terrestrial carnivores: Canidae, Felidae, Hyaenidae, Mustelidae, Procyonidae, Ursidae, and Viverridae. Veterinarians, including those working with nondomestic carnivores, should be familiar with the clinical signs, diagnosis, and clinical management of this disease. PMID- 11428392 TI - Infectious and parasitic diseases and contaminant-related problems of North American river otters (Lontra canadensis): a review. AB - The aquatic North American river otter (Lontra canadensis), a charismatic nonendangered mustelid, presently occupies a greatly reduced range. The species is common in zoos and has economic value as a furbearer. At least 17 states and one Canadian province have undertaken recent reintroduction programs. Nevertheless, little has been published on nonparasitic diseases of river otters, and little is known of the clinical significance of most of their parasitic infections. Records of environmental contaminant-related diseases and traumatic injuries in free-ranging North American river otters are also rare. This paper reviews present knowledge of North American river otter diseases, especially those with the greatest potential for impacting reintroduction programs. PMID- 11428393 TI - Clinical anatomy of the North American river otter (Lontra canadensis). AB - Published anatomic studies of North American river otters (Lontra canadensis) include only comparative reviews of single anatomic systems, but this paper includes multiple clinically relevant anatomic features. Both captive and free ranging North American river otters were studied by gross dissection of nine specimens, radiography of 18 live otters, and physical examination of 170 otters. Body fat in North American river otters is stored primarily subcutaneously at the base of the tail and in the axillae, so lack of visceral detail on abdominal radiographs is due to a lack of coelomic fat. The heart and great vessels are oriented as in the dog. The cardiac radiographic silhouette measures three intercostal spaces laterally between the fourth and sixth ribs and 4-4.5 intercostal spaces between the seventh through 11th ribs on the ventrodorsal projection. Subjective estimation of age by evaluation of tooth wear is confirmed by histologic examination of cementum annuli. Structures of the feet were imaged by high-resolution mammographic technique. The adult thymus was prominent, but the gland was small in the one juvenile animal studied. The spleen was unexpectedly large but consistent with findings in European otters (Lutra lutra). As in other otters and mammals, kidneys are reniculated. PMID- 11428394 TI - Hematology and serum biochemistry parameters of North American river otters (Lontra canadensis). AB - Blood samples were obtained from 155 North American river otters (Lontra canadensis; 94 adult males, 38 adult females, 10 juvenile males, and 13 juvenile females) to establish baseline hematology and from 50 adult river otters (40 males and 10 females) for baseline serum biochemistry parameters for the species. The otters were livetrapped from eastern North Carolina (USA) during a 4-yr period. Data for 14 routine hematologic parameters and 22 serum chemistry assays showed significant differences in total leukocyte count and absolute neutrophil and monocyte numbers for adults versus juveniles, red blood cell counts and hemoglobin between adult and juvenile males, and calcium and alkaline phosphatase values for adult males between years of the study and an increase in leukocyte counts and absolute neutrophils with increased degree of trap injury sustained. PMID- 11428395 TI - Hepatic iron accumulation over time in European starlings (Sturnus vulgaris) fed two levels of iron. AB - European starlings (Sturnus vulgaris) were used as a passerine bird model to examine the effect of dietary iron on the level of hepatic iron in birds. Nestling and fledgling starlings (n = 56) were raised on a controlled-iron diet. When birds maintained constant body weight, they were assigned in pairs to cages, and baseline sampling was performed. Pairs were then assigned to one of two diets: the controlled-iron diet (168 ppm, dry basis) or a high-iron diet (3,035 ppm, dry basis). Dry-matter intake and iron consumption were recorded. Dry-matter intake did not differ between the dietary treatment groups and was stable during treatment periods. Iron intake was higher in the high-iron group (P < 0.05). Birds were euthanized at baseline, 8 wk, and 16 wk. Body, liver, and spleen weights were measured. Hepatic iron and copper concentrations were determined. Body weight did not differ between the two treatment groups or among individuals for the study duration. Liver iron concentration differed over time and between treatment groups. Birds receiving both treatments had similar liver iron content at week 8 (3,107 +/- 228.6 ppm and 3,122 +/- 306.2 ppm high and controlled iron, respectively; P > 0.05), but by week 16, birds consuming the high-iron diet had greater hepatic iron levels than those consuming the controlled-iron diet (5,929 +/- 937.2 ppm and 3,683 +/- 229.5 ppm high and controlled iron, respectively; P < 0.05). Birds on the controlled-iron diet also had higher hepatic iron at 16 wk than at 8 wk. Liver copper decreased over time in all birds regardless of treatment. Results show that both dietary iron level and duration of time influenced hepatic iron storage. The controlled-iron diets still allowed accumulation of hepatic iron in an 8-wk period. PMID- 11428396 TI - Protein electrophoresis as a diagnostic and prognostic tool in raptor medicine. AB - Plasma proteins of 139 healthy adult birds of prey from 10 species were separated by electrophoresis to characterize and document normal reference ranges and species-specific electrophoretic patternsand to evaluate the value of this technique for health screening, disease diagnosis, and prognostic indication. Species studied included bald eagle (Haliaeetus leucocephalus), red-tailed hawk (Buteo jamaicensis), barn owl (Tyto alba), great horned owl (Bubo virginianus), turkey vulture (Cathartes aura), Harris' hawk (Parabuteo unicinctus), Stellar's sea eagle (Haliaeetus pelagicus), barred owl (Strix varia), screech owl (Otus asio), and black vulture (Coragyps atratus). Several clinical cases show the diagnostic/therapeutic value of protein electrophoresis in raptors. This study establishes species-specific reference ranges for several birds of prey and discusses the benefit of electrophoresis as a diagnostic technique in health screens, as a diagnostic aid in conjunction with other tests, and as a prognostic indicator in clinical evaluation of raptors. PMID- 11428397 TI - Identification and treatment of Moniliformis clarki (Acanthocephala) in cotton topped tamarins (Saguinus oedipus). AB - Ten cotton-topped tamarins (Saguinus oedipus) were infected with Moniliformis clarki a parasitic acanthocephalan. One animal died of intestinal perforation and peritonitis before treatment was initiated. Five of the remaining tamarins were treated successfully with oral albendazole at either 50 mg/kg b.i.d. for 16 days or 100 mg/kg b.i.d. for 3 days, then repeated biweekly for four treatments. A less-intensive treatment regime of 50 mg/kg albendazole orally s.i.d. for 3 days and monthly repeated treatments was unsuccessful. Parasites were removed via surgical enterotomy from two animals, both of which continued to shed acanthocephalan eggs postoperatively. PMID- 11428398 TI - Apparent digestibility of macronutrients in captive babirusa (Babyrousa babyrussa). AB - The apparent digestibilities of macronutrients by babirusa (Babyrousa babyrussa) were measured. Three group-housed captive babirusa were fed a diet of pelleted feed, boiled potatoes, grains, vegetables, and fruits. To determine reproducibility of the results, two digestibility trials were conducted 5 wk apart. The apparent digestibilities for crude protein, crude fat, and nonstructural carbohydrates (nitrogen-free extract) were 71, 76, and 78% of intake, respectively. Average apparent digestibilities of neutral detergent fiber and acid detergent fiber were 62 and 19% of intake, respectively. Hemicellulose (neutral detergent fiber minus acid detergent fiber) digestibility was 75%. Cellulose digestibility on average was 19% of intake, i.e., identical to acid detergent fiber digestibility because the diet did not contain a detectable amount of acid detergent lignin. PMID- 11428399 TI - Increasing the calcium content of mealworms (Tenebrio molitor) to improve their nutritional value for bone mineralization of growing chicks. AB - The purpose of these studies was to determine the husbandry variables that optimize the Ca content of mealworms (Tenebrio molitor) and to determine the bioavailability of this Ca for bone mineralization in chicks that consume the mealworms. To determine the optimal level of Ca in the substrates used in short term (< 14 days) holding of mealworms and to determine the length of time that mealworms should be exposed to high-Ca substrates, mealworms were placed in either a wheat bran or a chicken starter substrate supplemented with 0, 4, 8, or 12% Ca from CaCO3. The mealworms were harvested after 0.5, 1, 2, 3, 4, 7, or 14 days. The Ca content of the mealworms was greatest with the use of chicken starter and increased linearly with the Ca content of the substrate. In general, the Ca content of the mealworms increased during the first 24 hr and decreased after > or = 1 wk, especially at the higher levels of Ca supplementation. The chicken starter also resulted in higher levels of vitamin D in mealworms. Mealworms held in wheat bran with 8% Ca were fed to growing chicks. Ca bioavailability was calculated from the chicks' bone ash. The Ca in these mealworms was 76% as bioavailable as the Ca in oyster shell. PMID- 11428400 TI - Use of famciclovir for the treatment of endotheliotrophic herpesvirus infections in Asian elephants (Elephas maximus). AB - Two juvenile Asian elephants (Elephas maximus) presented with an acute onset of facial edema and lethargy. Examination of the oral cavity of each animal revealed cyanosis of the tip and distal margins of the tongue suggestive of endothelial inclusion body disease (EIBD) of elephants. Whole-blood samples were obtained, and polymerase chain reaction tests confirmed the presence of elephant herpesvirus. The animals were administered famciclovir (Famvir, SmithKline Beecham Pharmaceuticals, Philadelphia, Pennsylvania 19101, USA), a potent human anti-herpesvirus drug, in the course of their disease, and recovery followed a treatment regime of 3-4 wk. These are the first known cases of elephants surviving EIBD. PMID- 11428401 TI - Common cuttlefish (Sepia officinalis) mortality at the National Zoological Park: implications for clinical management. AB - Six out of seven cuttlefish acquired by the Smithsonian National Zoological Park in July 1998 died before 1 November 1998. Postmortem examinations showed mantle ulcers, secondary bacterial infections, inanition, and cuttlebone fractures. The surviving cuttlefish developed a progressive focal mantle ulcer, was treated with oral chloramphenicol intermittently for 9 wk, and maintained a normal appetite and growth rate until death at 7 mo of age. The National Zoological Park pathology database showed signalments, histories, and causes of mortality of 186 common cuttlefish, each 1-14 mo old, that received gross and histologic examinations; for example, the largest group of cuttlefish of known sex, age, and body weight at postmortem were 7-9 mo old and weighed an average of 376.2 g (males, n = 18) and 299.0 g (females, n = 15). Many cuttlefish had multiple pathologic diagnoses. Significant diseases included inflammation and secondary bacterial infections, especially gastrointestinal, cardiovascular, respiratory, reproductive, and ophthalmic, and septicemia due to Vibrio spp. or other gram negative bacteria. Mantle lesions, including ulceration/dermatitis, abscess/granuloma, necrosis/fibrosis/cellulitis, and laceration/abrasion/erosion, were also identified, along with inanition, cuttlebone lesions, and trauma. Mantle lesions were associated with secondary bacterial infections and death. On the basis of this information, if captive cuttlefish behavior creates risk for development of mantle lesions, administration of antibiotics effective against gram-negative bacteria may delay or halt disease progression. Cuttlefish exhibits require proper design, husbandry, economic resources, and staffing to minimize disease syndromes and mortality. PMID- 11428402 TI - Chorionic villus sampling for sex determination in a western lowland gorilla (Gorilla gorilla gorilla). AB - Chorionic villus sampling was undertaken on an anesthetized gorilla (Gorilla gorilla gorilla) to determine the fetal sex in the first trimester of pregnancy. The tissue samples were subject to sex determination by polymerase chain reaction, fluorescent in situ hybridization, and cell culture with cytogenetic analysis. Polymerase chain reaction testing was found to be the most accurate and rapid method of sex determination on this tissue sample. PMID- 11428403 TI - Management of reactive arthritis in a juvenile gorilla (Gorilla gorilla gorilla) with long-term sulfasalazine therapy. AB - A juvenile western lowland gorilla (Gorilla gorilla gorilla) experienced recurrent fever, lethargy, diarrhea, and/or arthritis starting at age 6 mo. During an episode at age 15 mo, Shigella sp. was isolated from diarrheic feces. At age 41 mo, reactive arthritis was diagnosed. In addition, the gorilla's growth was retarded. All arthritic attacks were managed symptomatically prior to age 4 yr, at which time a severe episode precipitated the implementation of therapy with sulfasalazine, an arthritis suppressive medication. Examination 27 mo later revealed cessation of progressive joint pathology although the animal exhibited decreased range of motion in most joints. The gorilla has been on sulfasalazine therapy for 4 yr without lameness. Growth has resumed, and there has been no radiographic evidence of progressive joint degeneration. Immunogenetic analysis of whole blood obtained at age 68 mo identified the gorilla major histocompatibility class I allele, Gogo-B*0101, which has limited nucleotide sequence similarity to HLA-B27, an allele associated with postinfection reactive arthritis in humans. Sulfasalazine therapy effectively managed reactive arthritis in this gorilla and should be considered for similarly frequently affected animals. Juvenile gorillas, in populations with a history of clinical shigellosis and/or postdiarrhea arthritis, may benefit from prophylactic sulfasalazine therapy after episodes of bacterial enterocolitis. Sulfasalazine therapy should be considered in all gorillas, juvenile and adult, experiencing confirmed Shigella sp.-associated enterocolitis. PMID- 11428404 TI - Use of pyloroplasty (Y-U) to treat presumed delayed gastric emptying in a cheetah (Acinonyx jubatus). AB - A 4-yr-old cheetah (Acinonyx jubatus) with a 2-yr history of chronic intermittent vomiting and spiral bacteria-associated gastritis presented with dramatically increased vomiting frequency and marked intermittent abdominal distention. Physical examination revealed loss of muscle mass and poor fur coat quality. Contrast radiography was consistent with delayed gastric emptying due to presumed gastric outlet obstruction. Both Y-U pyloroplasty and incisional gastropexy were performed, and no subsequent vomiting has been observed for 3 yr with the exception of three episodes during the immediate postoperative period. The cause of delayed gastric emptying was not determined, although a gastric motility disorder associated with gastric bacterial infection and elevated gastrin levels was suspected. PMID- 11428405 TI - Tyzzer's disease in a red panda (Ailurus fulgens fulgens). AB - A debilitated 9-yr-old female red panda (Ailurus fulgens fulgens) with a recent history of corticosteroid administration displayed anorexia, depression, and diarrhea for 2 days. Blood work revealed a moderate nonregenerative anemia, leukocytosis, hypokalemia, hyperbilirubinemia, and mildly elevated alanine aminotransferase and aspartate aminotransferase. Serology was negative for occult heartworm, Toxoplasma gondii, feline leukemia virus, feline infectious peritonitis, feline immunodeficiency virus, and canine distemper virus. Electron microscopy of the feces demonstrated corona-like virus particles. The panda died 3 days after initial presentation. Histologic findings included multifocal, acute, hepatic necrosis and diffuse, necrotizing colitis. Liver and colon lesions contained intracellular, curved, spore-forming, gram-negative, silver-positive rods morphologically consistent with Clostridium piliforme. This panda most likely contracted Tyzzer's disease subsequent to having a compromised immune system after corticosteroid administration and concurrent disease. PMID- 11428406 TI - Glucagonoma in a jaguar (Panthera onca). AB - An obese adult jaguar (Panthera onca) was euthanized because of progressive lameness. Two 3-cm-diameter pancreatic nodules were identified as islet cell tumors, which were positive with immunohistochemical stains for glucagon, neuron specific enolase, chromogranin A, and synaptophysin. The jaguar did not present clinical evidence of hyperglucagonemia. PMID- 11428407 TI - Birth of Siberian tiger (Panthera tigris altaica) cubs after transvaginal artificial insemination. AB - A captive 9-yr-old female Siberian tiger (Panthera tigris altaica) at the Lisbon Zoo was treated with equine chorionic gonadotropin followed 80 hr later by human chorionic gonadotropin to stimulate follicular development and ovulation, respectively. Forty-one hours after the second hormone administration, the tiger was subjected to transvaginal artificial insemination with diluted fresh semen containing 500 x 10(6) motile spermatozoa obtained from an 8-yr-old male Siberian tiger via electroejaculation. After anesthesia, the female tiger was placed in an inclined position with the hind limbs elevated during and after the insemination and was maintained in this position for 15 min to minimize semen reflux. An anesthetic reversal agent was then administered. After a 103-day gestation, the female gave birth to three apparently healthy cubs that survived 24-48 hr. These results demonstrate that transvaginal artificial insemination can be successfully performed in the Siberian tiger to produce full-term offspring. PMID- 11428408 TI - Trombiculid mites (Hannemania sp.) in canyon tree frogs (Hyla arenicolor). AB - Six wild-caught, captive canyon tree frogs (Hyla arenicolor) purchased as a group and housed at the North Carolina Zoological Park were diagnosed with intradermal Hannemania mites, the first described infestation of this frog species by this mite. Typical gross lesions were orange-colored skin vesicles approximately 1 mm in diameter, predominantly located on the ventrum and ventral hind limbs. The larval mites had ovoid bodies approximately 44 microm in length and 240 microm in width. Three of the frogs died, and three became free of mites and pathologic changes after approximately 28 wk of treatment with transcutaneous ivermectin and manual mite removal. The frogs remained free of mites at a 1-yr recheck. PMID- 11428409 TI - Cat flea (Ctenocephalides felis) infestation in quarantined marsh deer (Blastocerus dichotomus) populations. AB - Marsh deer (Blastocerus dichotomus) were captured for a research program in Brazil and maintained in quarantine stations. After 60 days, fleas were detected on animals and identified as Ctenocephalides felis felis. Elimination of the infestation was difficult. Animal treatment with a fipronil-based compound was effective, and subsequently captured animals were treated prophylactically. Some animals remained infested, and some died from the infestation. PMID- 11428410 TI - Clinical challenge. Metallic foreign body in the region of the proventriculus. PMID- 11428411 TI - Clinical challenge. Oral mucosa, tongue, gingiva, lips; multiple papillomas (oral papillomatosis). PMID- 11428412 TI - The radiology of pulmonary cryptococcosis in a tertiary medical center. AB - Pulmonary cryptococcal infections occur in both immunocompetent and immunocompromised individuals, with a reported increased incidence of diffuse pulmonary disease in acquired immune deficiency syndrome (AIDS) patients. The authors observed no differences in the radiographic appearances of pulmonary cryptococcal disease between human immunodeficiency virus (HIV) patients and other immunocompromised individuals. Chest computed tomography (CT) contributes to a more comprehensive understanding of pulmonary cryptococcal infections. PMID- 11428413 TI - Diaphragmatic shape change after lung volume reduction surgery. AB - Diaphragmatic shape in normal patients was significantly different from shape in emphysema patients. Postoperative diaphragmatic shape in patients with good clinical outcome differed from preoperative shape and was similar to shape in normal patients. In patients with poor clinical outcome, surgery appeared to have little effect on diaphragm shape. PMID- 11428414 TI - Large cell neuroendocrine carcinoma of the lung: clinical, CT, and pathologic findings in 11 patients. AB - The purpose of this study was to describe the clinical, computed tomographic (CT), and pathologic findings of large cell neuroendocrine carcinoma (LCNEC) of the lung. CT and pathologic findings as well as clinical features of surgically proven LCNEC of the lung were reviewed retrospectively in 11 consecutive patients (eight men and three women; mean age, 63 years; range, 44-77 years). Chest CT showed peripheral mass or nodule (n = 8) and central mass with distal atelectasis (n = 3). Six tumors were accompanied by mediastinal (n = 3) and hilar (n = 3) lymph node enlargement at CT. On pathologic examination, all resected tumors showed necrosis of variable extent (mean: 38%, range; 10-70%). The areas of intrinsic lipoid pneumonia and tumor emboli in two patients appeared at CT as areas of ground-glass opacity surrounding the tumor. Mediastinal nodal metastases were seen in three (27%) patients. Pathologic staging of 11 patients was IB in six, IIA in one, IIB in one, IIIA in two, and IIIB in one. Follow-up data showed extrathoracic metastases in four patients at mean follow-up period of 15 months. One patient died of distant metastasis 5 months after the surgery. CT findings of LCNEC of the lung are nonspecific and similar to those of other non-small cell lung cancers and extrathoracic metastasis is seen in approximately one third of the patients with follow-up study. PMID- 11428415 TI - The effect of imaging modality on patient management in the evaluation of pulmonary thromboembolism. AB - A retrospective medical record review was performed to study the differences in clinical risk profiles and the relationships between test results versus management for suspected pulmonary thromboembolism (TE) in patients undergoing either radionuclide ventilation perfusion (V/Q) scans or pulmonary computed tomographic angiography (CTA), as the initial test. Data of 138 consecutive V/Q patients were compared with that of 149 consecutive CTA patients during equivalent 6-month intervals before and after the introduction of CTA. Information on risk factors, signs and symptoms, all diagnostic test results, and the relationships between the test results and ultimate physician management were collected and analyzed. V/Q results predicted physician management in all patients with high probability scans and 91% with normal to low probability scans. There were 35 patients with indeterminate V/Q scans--43% of these patients were managed without any other diagnostic test. CTA results predicted management in all patients with positive studies and 99% of patients with negative studies. In contrast to the V/Q cohort, only seven CTA studies were inconclusive- additional diagnostic tests determined management in all but one case. Compared with V/Q, CTA has fewer indeterminate results, is more directly reflective of management, and reduces the number of patients managed with inconclusive data. PMID- 11428416 TI - Right hilar pseudomass due to partial right lower lobe atelectasis. AB - In cases of right lower lobe atelectasis, in which the lobe remains partially aerated, rotation of the right hilum into an anteroposterior plane can result in the formation of a "pseudomass" in the right hilar region on the frontal radiograph. Failure to recognize this variant appearance of right lower lobe atelectasis can result in a false diagnosis of a hilar mass and result in an unnecessary workup for lung cancer. Familiarity with this variant appearance of right lower lobe atelectasis, coupled with careful comparison with recent normal chest radiographs, should allow a correct diagnosis of a "pseudomass" in most cases. Computed tomography of the thorax can reliably exclude a hilar mass in cases in which recent normal chest radiographs are unavailable for comparison. PMID- 11428417 TI - Gallstone empyema complicating laparoscopic cholecystectomy. AB - Empyema may rarely present as a delayed complication after laparoscopic cholecystectomy. Patients with this complication invariably have associated dropped gallstones in the peritoneal cavity. The gallstones may erode through the diaphragm or migrate through preexisting defects in the diaphragm. The latter are seen in over 50% of the elderly population and may predispose them to this rare complication. PMID- 11428418 TI - Primary diffuse tracheobronchial amyloidosis: radiologic findings. AB - Reports detailing the radiologic features of tracheobronchial amyloidosis are scant and typically describe only the computed tomography findings of this disease. No reports have described the magnetic resonance (MR) findings of tracheobronchial amyloidosis. With this case, the authors describe the radiologic findings in a patient with diffuse tracheobronchial amyloidosis, focusing particularly on the MR and bone scintigraphy findings in this disease. PMID- 11428419 TI - Malignant peripheral nerve sheath tumors of intrathoracic vagus nerve. AB - The authors report two cases of malignant peripheral nerve sheath tumors arising from the vagus nerve in the mediastinum in patients who had no stigmata of neurofibromatosis (von Recklinghausen's disease). Computed tomography showed homogeneous soft tissue masses, with minimal calcification seen in one patient. T2-weighted magnetic resonance images showed regions of low signal intensity caused by a dense cellular population. PMID- 11428420 TI - Pulmonary lymphangiomyomatosis mimicking Langerhans cell histiocytosis. AB - The authors describe a patient with histologically confirmed pulmonary lymphangiomyomatosis and thin-section computed tomography findings mimicking Langerhans cell histioctytosis. The description emphasizes the nonspecificity of the computed tomography findings in this patient. This report also suggests that the computed tomography diagnosis of lymphangioleiomyomatosis can be difficult at an early stage of the disease and should be complemented by biopsy verification. PMID- 11428421 TI - Hemophilic pseudotumor of the chest. AB - We report the progressive imaging findings of a case of hemophilic pseudotumor of the chest, not previously described. The pseudotumor, although originally extrapleural, has gradually enlarged and eroded into a bronchus, producing a bronchopleural fistula. PMID- 11428422 TI - Tuberculous fibrosing mediastinitis: CT and MRI findings. AB - Fibrosing mediastinitis is a rare entity usually caused by granulomatous disease. Most cases develop as a complication of histoplasmosis. Mediastinal involvement caused by tuberculosis most often produces focally enlarged lymph nodes, with diffuse mediastinal involvement a very unusual manifestation of disease. The authors describe a patient with extensive tuberculous fibrosing mediastinitis, in whom fibrotic masses produced compression of airways and vascular structures with resultant parenchymal changes. PMID- 11428423 TI - Friendship quality and sociometric status: between-group differences and links to loneliness in severely abused and nonabused children. AB - OBJECTIVE: There were two main aims: first, to illuminate the difference between abused children's general popularity with classmates and success in close friendships; second, to examine the specific interactional qualities of abused children's friendships and their links to loneliness. METHOD: Thirty-five severely abused children and 43 matched, nonabused children were compared on peer rated sociometric status, self-reported loneliness, and observed and self reported friendship quality. RESULTS: Abused children were not rated significantly lower sociometrically, nor did they differ significantly from control children on several measures of friendship quality, such as resolving conflicts and helping each other. However, abused children were observed to be more negative and less proactive in their interactions. They also reported their friendships as being more conflictual, and as higher on betrayal and lower on caring. Only observational friendship variables predicted loneliness. CONCLUSIONS: The results challenge the assumption that abused children's peer relationships are uniformly more maladaptive than nonabused children's, and point to the possible benefits of structured interventions for "normalizing" their friendship interactions. The pattern of difficulties exhibited by abused children (e.g., conflict) provides foci for more specific interventions. Multi-method assessments are necessary and the multi-dimensional nature of children's social adjustment is important to understand. PMID- 11428424 TI - A comparative study of risk assessments and recommended interventions in Canada and Israel. AB - OBJECTIVES: Our goal was to examine how professionals assess children at risk and their parents, and decide on particular interventions. Specifically, we explored whether their assessments and decision-making are influenced by (1) the mother's degree of cooperativeness and/or (2) the country in which the worker lives (Canada or Israel). METHOD: Workers working in the child welfare field (N = 181) were presented with a case vignette and asked to assess the child and parents, and the degree of risk to the child, and make an intervention recommendation. The measures used in this study were based on previous work and field-tested in both countries. RESULTS: Significant differences were found between the two countries regarding workers' age and level of experience, with Canadians being older and more experienced than Israelis. Significant differences were also found between the two countries regarding the assessments of the child and parents and also of risk to the child, with Canadians assessing significantly more stringently than Israelis. The difference in levels of experience between the two countries did not explain these differences; however, it did influence intervention recommendations, only for those with 3 years or more of experience. Within this group, significantly more Canadians than Israelis recommended removing the child from the home. Regarding maternal cooperativeness, this factor did affect workers' assessments of the mother, but not of the father or child, or the worker's recommended intervention. Israelis' assessments were significantly more influenced by the mother's cooperativeness than Canadians'. CONCLUSIONS: Significant differences were found between the Canadian and Israeli professionals in this study in both their assessments and their intervention recommendations. These appear to reflect the different social, cultural, and political contexts in which these professionals work, and underscores the value of cross-national comparative studies in child welfare. PMID- 11428425 TI - Child emotional maltreatment: a 2-year study of US Army cases. AB - OBJECTIVE: Reports of childhood emotional maltreatment have increased greatly over the past decade. The objective of this research was to determine the types of emotional maltreatment substantiated in a community of US Army families residing temporarily in Germany. Such a description may help to improve the understanding of how a jurisdictional body defines emotional maltreatment in day to-day practice. METHOD: Data were obtained from a review of the minutes of case review committees (CRCs) for 181 cases of child emotional abuse in 1997-1998. We determined the type, number, and severity of incidents, the substantiation rate, and the situations to which children were exposed. RESULTS: The most frequently substantiated type of incident was witnessing domestic violence, 60% of all cases. Primary emotional abuse was found in 26% of cases, while emotional abuse in conjunction with child physical abuse or child neglect was found in 14% of cases. The more severe the case, the more likely it was to be substantiated. CONCLUSIONS: Emotional maltreatment was substantiated more as a single type than in combination with other forms of maltreatment. Seeing emotional abuse as a single entity may allow clinicians to focus on a relationship or situation (such as spouse abuse) that is potentially harmful to a child. However, recognizing the emotionally abusive aspects of child physical abuse and neglect could allow an expanded treatment plan that could include treatment of the emotionally abusive behavior to strengthen the relationship of the caregiver to the child, in addition to the focus on the physical abuse and neglect. PMID- 11428426 TI - Lay therapy intervention with families at risk for parenting difficulties: The Kempe Community Caring Program. AB - OBJECTIVE: To determine which areas of family functioning lay therapy intervention can improve in a home visitation program for first time mothers at high risk for parenting difficulties, including the potential for child abuse and neglect. METHODS: Families were assessed for risk of parenting difficulties using the Parent Readiness and Risk Assessment Checklists. Lay therapists completed the Scale of Family Functioning pre- and postintervention with 108 high risk families for whom they provided support, education, and referrals. RESULTS: Paired t tests were done to determine whether the eight categories of the Scale of Family Functioning improved from baseline to termination of the service. Four categories showed statistically significant improvement: social support (p = .001), self esteem (p < .001), confidence as a parent (p < .001), and affective relationships (p = .003). CONCLUSIONS: Statistically significant improvement occurred in four of the eight categories on the Scale of Family Functioning indicating that home intervention programs can improve some categories of family functioning. Longer intervention and an increased focus on improving parent/infant interactions may be able to increase parental sensitivity and expectations. In this study, the categories of family conflict and stability/meeting basic needs were not amenable to change with lay therapy intervention. PMID- 11428427 TI - Child protective services' response to prenatal drug exposure: results from a nationwide survey. AB - OBJECTIVE: This survey was conducted to: (1) document child protective services (CPS) agencies' actual practices regarding prenatal drug exposure; (2) examine urban and rural differences in CPS responses; (3) explore whether CPS practices varied as a function of county median income, birth rate, population size, or percent minority births; and (4) assess respondent satisfaction with their county's current responses. METHOD: Data were collected via a nationwide telephone survey of child welfare supervisors from two urban and two rural counties in every state (N = 200). RESULTS: Ninety percent of counties (100% of urban and 80% of rural) reported receiving referrals of infants with prenatal drug exposure. Among those receiving referrals, extreme variations in practice were found; all possible response options (from very inactive to very aggressive) were equally represented on key questions (e.g., filing court petitions, taking custody). Rural counties tended to have stronger responses than urban counties (t[175] = -2.26, p = .024). County response did not vary with county-level median family income, percent minority births, or birth rate. Despite wide variations in practice, the majority of respondents (69%) felt their county's response was appropriate. Of respondents who did indicate that their county's response was inappropriate, most (85%) felt that the county needed to do more to protect children. CONCLUSIONS: There is currently tremendous variation across US counties in CPS responses to, and beliefs regarding, the issue of prenatal drug exposure. Some of this variation is due to differences between urban and rural counties, with little variation explained by differences in median income or percent minority births at the county level. There is a need for research-based guidance and consensus building in CPS practice in this area. PMID- 11428428 TI - The Memorandum of Good Practice: theory versus application. AB - OBJECTIVE: The goal of this study was to evaluate the quality of investigative interviews in England and Wales since implementation of the Memorandum of Good Practice (MOGP), which specified how forensic interviews of alleged child abuse victims should be conducted. METHOD: Transcripts of 119 videotaped interviews of alleged victims between the ages of 4 and 13 years were obtained from 13 collaborating police forces. Trained raters then classified the types of prompts used by the investigators to elicit substantive information from the children, and tabulated the number of forensically relevant details provided by the children in each response. RESULTS: Like their counterparts in the United States, Israel, and Sweden, forensic interviewers in England and Wales relied heavily on option-posing prompts, seldom using open-ended utterances to elicit information from the children. Nearly 40% of the information obtained was elicited using option-posing and suggestive prompts, which are known to elicit less reliable information than open-ended prompts do. CONCLUSION: Despite the clarity and specificity of the MOGP, its implementation appears to have had less effect on the practices of forensic interviewers in the field than was hoped. Further work should focus on ways of training interviewers to implement the superior practices endorsed by the MOGP and similar professional guidelines. PMID- 11428429 TI - On the interpretation of pictures with and without a content of child sexual abuse. AB - OBJECTIVE: The purposes of the three experiments were to validate the possibility of a picture to evoke the recognition of child sexual abuse, to determine if the picture was anxiety evoking, and to investigate if the content of child sexual abuse would be transferred to a neutral picture. METHOD: In all three experiments, adult men and women were presented with a drawing intended to depict child sexual abuse, and were requested to interpret the picture. Experiment 1: Before and after the picture presentation, 226 participants were given a test of anxiety. Experiment 2: After the exposure of the child abuse picture, 200 new participants were asked to interpret an innocent child-adult picture. Experiment 3: To complete Experiment 2, 89 new participants were asked to interpret the pictures in the reverse order. RESULTS: Almost three-fourths of the participants saw child sexual abuse in the picture with a sexual threat. Those in Experiment 1 who saw the picture as child sexual abuse or as a problematic child-adult situation without sexual implications reported a significant increase of anxiety level. None in Experiment 2 or 3 saw child sexual abuse in the innocent picture. The sex of the abused child was significantly more often interpreted as opposite to one's own sex. CONCLUSIONS: The study indicates some people's deficient capacity to recognize the message of child sexual abuse in the picture. It seems that certain people can spare themselves anxiety by not registering the child's precarious situation or not seeing the child as being of their own sex. This has implications for the recognition of child sexual abuse in society. It was also shown that a sexual abuse theme was not transferred from one context to another context, which immediately followed it. PMID- 11428430 TI - Classification models of child molesters utilizing the Abel Assessment for sexual interest. AB - OBJECTIVE: The aims of this study are to demonstrate 1) the criterion validity of the Abel Assessment for sexual interest (AASI) based on its ability to discriminate between non child molesters and admitting child molesters, and 2) its resistance to falsification based on its ability to discriminate between liar denier child molesters and non child molesters. METHOD: A group of 747 participants matched by age, race, and income was used to develop three logistic regression equations. The models compare a group of non child molesting patients under evaluation for other paraphilias to three groups: 1) a group of admitting molesters of girls under 14 years of age, 2) a group of admitting molesters of boys under 14 years of age, and 3) a group believed to be concealing or denying having molested. RESULTS: Both of the equations designed to discriminate between admitting child molesters and non child molesters were statistically significant. The equation contrasting child molesters attempting to conceal or deny their behavior and non child molesting patients was also statistically significant. CONCLUSIONS: The models classifying admitting child molesters versus non child molesters demonstrate criterion validity, while the third model provides evidence of the AASI's resistance to falsification and its utility as a tool in the detection of child molesters who deny the behavior. Results of the equations are reported and suggestions for their use are discussed. PMID- 11428431 TI - Outcomes of child sexual contacts: patterns of incarcerations from a national sample. AB - OBJECTIVE: In a test of the relationships between sexual touching before puberty and later incarcerations during adolescence and adulthood, two theoretical models were examined. These models focus on examination of crime from the developmental origins of criminals and the adaptive and maladaptive outcomes of early experiences. METHOD: Data were taken from the National Health and Social Life Survey to study the sexual touching and social origin variables in childhood and incarceration variable during teen years. The sample was a stratified, random sample of 3,362 adults (18 to 59 years of age) throughout the United States. RESULTS: Data support inferences from the two interrelated models. Respondents reporting touching before puberty were significantly more likely to experience incarceration. They were also more likely to engage in behaviors as teenagers that were associated with an increased probability of jailing. Such behaviors include sexual promiscuity and early departure from the parental family. CONCLUSIONS: These findings are consistent for first jailing as a teenager and as an adult, as well as for those with short (less than a week) or longer (a week or more) spells of incarceration. Prepubertal sexual touching has a significant and enduring effect on later adolescent and adult incarcerations. PMID- 11428432 TI - Glycopeptide resistance in Staphylococcus haemolyticus during treatment with teicoplanin. PMID- 11428433 TI - Alarming baseline rates of nosocomial infection and surgical prophylaxis errors in a small teaching hospital in Argentina. PMID- 11428434 TI - Surgical-site infections (SSI) and the NNIS Basic SSI Risk Index, part II: room for improvement. PMID- 11428435 TI - Suitability of the NNIS index for estimating surgical-site infection risk at a small university hospital in Brazil. AB - OBJECTIVES: To detect the occurrence of surgical-site infection (SSI) in our study sample, using the traditional variables of the National Nosocomial Infection Surveillance (NNIS) index with a locally modified cut-off point for the "T time" defining length of surgical procedure; to compare the modified and the traditional NNIS index under the hypothesis that a cut-off point discriminating procedures of short and long duration, based upon the actual experience of the study sample, can adequately predict the risk of SSI. DESIGN: A retrospective chart review of 9,322 patients undergoing surgical procedures in the period January 1993 to December 1998. SETING: A small university hospital (UH) in southern Brazil. RESULTS: The composite index using the local sample procedure duration cut-off point (UH-index) performed better than any of the individual components of the composite index (anesthesia risk index and surgical-wound class [SWC]). The UH-index also predicted adequately the risk of SSI when compared to the traditional NNIS index, particularly when stratifying by SWC. CONCLUSIONS: A modified NNIS index, using the sample cut-off point, can adequately predict the risk of SSI in a given population. Further studies are needed to compare and validate the NNIS index of risk for populations other than those of the NNIS participating hospitals. Larger samples using different hospitals with similar characteristics are needed to investigate the risk of SSI associated with specific operations. PMID- 11428436 TI - Assessing excess nurse work load generated by multiresistant nosocomial bacteria in intensive care. AB - OBJECTIVE: To compare three methods for assessing the excess nurse work load related to recommended procedures for managing nosocomial infections (NI) due to multiresistant bacteria (MRB): two activity scores, the Omega score and the Projet de Recherche en Nursing (PRN) system, and a specific evaluation based on functional analysis of nursing procedures. SETTING: 10 beds in a medical intensive care unit (MICU). PATIENTS: Patients admitted from November 15, 1995, to June 15, 1996, were included and divided in two groups based on presence of MRB colonization or infection (MRB+ and MRB-groups). METHODS: Data were collected regarding length of stay (LOS) in days; Omega score for the entire stay; PRN score for the entire stay and per day; and time required to perform correctly four nursing procedures related to MRB NI, as evaluated specifically by the nursing staff, using a detailed functional analysis document that described all elementary nursing tasks in chronological order and all material needed to carry out those tasks. RESULTS: The LOS and total Omega and PRN scores were higher in the MRB+ group than in the MRB- group: LOS, 23 +/- 20.6 versus 12 +/- 15.3 days, (P<.001); Omega score, 164 +/- 103.4 versus 123 +/- 93.7 points (P<.001); PRN score, 3,606 +/- 3,187 versus 1,854 +/- 2,356 points (P<.001), respectively. The daily PRN score was also higher in MRB+ group (PRN, 160 +/- 25 vs 146 +/- 34 points in the MRB- group; P<.028). Four nursing procedures made necessary by MRB acquisition were identified: isolation precautions, with two levels according to whether the risk of contamination was mild-moderate or high; bathing the patient with antiseptic solution; bedpan management; and microbiological screening. The functional analysis indicated that the time needed to carry out these four procedures correctly was 245 minutes per patient per day, as compared to 85 minutes according to the PRN system. CONCLUSIONS: Our data confirm that MRB NIs are responsible for an increase in nurse work load, as estimated by LOS, Omega, and PRN scores. However, the daily excess nurse work load related directly to recommended procedures for managing MRB NIs in MICUs is underestimated by these activity scores, as compared to a specific functional analysis of nursing tasks. This may be of importance in evaluating potential links between nurse work load and MRB NIs and in determining the number of nurse hours needed to comply with infection control recommendations. PMID- 11428437 TI - Persistence of immunity to varicella-zoster virus after vaccination of healthcare workers. AB - OBJECTIVE: Varicella-zoster virus (VZV) vaccine is recommended to protect susceptible healthcare workers (HCWs) from serious disease and to prevent nosocomial spread of VZV. We evaluated clinical outcomes and serological responses in HCWs after immunization with live attenuated VZV vaccine. DESIGN: Vaccinees were immunized from 1979 to 1998 during VZV vaccine trials, as well as after licensure, and followed prospectively for 1 month to 20.6 (mean 4.6) years after vaccination. Sera were tested by fluorescent antibody to membrane antigen (FAMA), latex agglutination (LA), and enzyme-linked immunoassay (EIA) to detect VZV-specific antibodies. STUDY PARTICIPANTS: The median age of the 120 HCWs was 26 years; 51 (42%) were males. INTERVENTIONS: Ninety eight (82%) of these study subjects received vaccine prepared by Merck and 22 (18%) by SmithKline Beecham; 25, 81, and 14 vaccinees received one dose, two doses, and three doses, respectively. RESULTS: The crude attack rate was 10%; 12 of 120 HCWs developed chickenpox 6 months to 8.4 years after vaccination. The attack rates following household and hospital exposures were 18% (4/22) and 8% (6/72), respectively. All resulting illness was mild to moderate (mean 40 vesicles). Seroconversion after vaccination was documented by FAMA in 96% of HCWs, although 31% lost detectable antibodies. Compared with FAMA, LA and EIA were 82% and 74% sensitive and 94% and 89% specific, respectively. CONCLUSIONS: The VZV vaccine effectively protected HCWs from varicella, particularly from serious disease. Currently available serological tests are not optimal, and improved assays are needed. PMID- 11428438 TI - Antibiotic resistance is a major risk factor for epidemic behavior of Acinetobacter baumannii. AB - OBJECTIVE: To study the presence of bacterial factors in clinical isolates of Acinetobacter species in order to identify markers of epidemic potential. DESIGN: Case-control study. METHODS: Forty-six isolates of Acinetobacter species, including 23 epidemic and 23 sporadic strains from different outbreaks in nine European countries, were compared for the presence of the following factors: hemagglutination, presence of capsules and fimbriae, binding to salivary mucins, resistance to drying, and antibiogram typing. Genotyping of all strains was performed by amplified fragment-length polymorphism (AFLP). RESULTS: All outbreak strains except two (91%) were identified as Acinetobacter baumannii. Binding to salivary mucins and resistance to antibiotics were significantly associated with epidemic behavior. Antibiogram typing showed clustering of predominantly A baumannii strains within one group, and these strains were significantly more resistant to antibiotics than sporadic strains. AFLP genotyping revealed a great heterogeneity among the different European Acinetobacter strains. Cluster analysis of AFLP fingerprints showed several small clusters of different A baumannii outbreak strains. AFLP genotyping could not identify a common epidemic marker within the strains studied. CONCLUSIONS: Antibiogram typing can be used in routine clinical laboratories as a screening method to recognize potentially epidemic A baumannii strains. Several other factors were found, both in different outbreaks as well as in sporadic Acinetobacter isolates. These characteristics were unable to predict epidemic behavior and therefore cannot be used as discriminative epidemic markers. AFLP genotyping demonstrated no common clonal origin of European epidemic A baumannii strains. This indicates that any clinical A baumannii isolate with resistance to multiple antibiotics can be a potential nosocomial outbreak strain. PMID- 11428439 TI - Emergency department management of occupational exposures: cost analysis of rapid HIV test. AB - OBJECTIVE: To compare costs for evaluation and treatment of a healthcare worker (HCW) experiencing an occupational exposure, using a rapid human immunodeficiency virus (HIV) test versus a standard enzyme-linked immunosorbent assay (ELISA) HIV test. DESIGN: Retrospective chart review of all HCWs presenting to the emergency department (ED) for care of an occupational exposure over a 13-month period. SETTING: A 404-bed university-based level 1 trauma center with an annual ED census of approximately 35,000. PARTICIPANTS: All HCWs experiencing an occupational exposure treated in the ED using a rapid HIV protocol were included in the analysis. METHODS: A calculation of selected costs of the initial evaluation and treatment of patients whose evaluation included a rapid HIV test on the source patient were performed. A similar calculation was then made for these patients, had the standard ELISA test been used. Evaluated costs included laboratory tests, postexposure prophylactic medications, and estimated lost work time. Other costs were constant and were not included in the evaluation. RESULTS: Total evaluated cost using the rapid HIV test as part of the evaluation and treatment protocol was $465.80 for 17 patients. Had the ELISA test been used instead of the rapid test, the total evaluated cost for the 17 patients would have been $5,965.81. CONCLUSIONS: When used as part of the evaluation and treatment of the HCW with an occupational exposure, the rapid HIV test results in substantial cost savings over the ELISA test . PMID- 11428440 TI - Rapid molecular typing of methicillin-resistant Staphylococcus aureus by PCR RFLP. AB - OBJECTIVE: To establish a new, rapid, and reliable genotypic fingerprinting technique for methicillin-resistant Staphylococcus aureus (MRSA) typing in routine epidemiological surveillance. DESIGN: The method is based on polymerase chain reaction (PCR) restriction fragment-length polymorphism (RFLP) following HaeII digestion of simultaneously amplified parts of the protein A gene, the coagulase gene, and the hypervariable region adjacent to mecA. A total of 46 MRSA initial isolates were analyzed, including 14 isolates from five countries; the six German epidemic strains; 16 isolates from the Frankfurt metropolitan area, which were known to be heterogeneous by pulsed-field gel electrophoresis (PFGE); and 10 isolates obtained during three epidemics, all of which displayed an identical genotype. RESULTS: Restriction analysis by PCR-RFLP permitted discrimination of 10 of 14 international isolates, all six German epidemic strains, and 15 of 16 national isolates. It also confirmed the homogeneous character of the 10 outbreak isolates. CONCLUSIONS: This new and rapid PCR-RFLP typing method is an attractive tool in routine epidemiological surveillance. Its impressive characteristics are ease of performance and interpretation, while at the same time guaranteeing good discriminatory power, reproducibility, and typeability. PMID- 11428441 TI - Nosocomial rotavirus in a pediatric hospital. AB - We describe a nosocomial rotavirus outbreak among pediatric cardiology patients and the impact of a prospective, laboratory-based surveillance program for rotavirus in our university-affiliated, quartenary-care pediatric hospital in New York City. Improved compliance with infection control and case-finding among patients and healthcare workers halted the outbreak. PMID- 11428442 TI - Outbreak of vancomycin-resistant Enterococcus faecium in a neonatal intensive care unit. AB - An outbreak of vancomycin-resistant Enterococcus faecium involving 28 infants in a neonatal intensive care unit was observed. Successful control of the outbreak was achieved following use of patient and staff cohorting, contact isolation precautions, patient and environmental surveillance cultures, environmental decontamination, molecular typing, introduction of an alcohol-based hand disinfectant, and decreased use of vancomycin. PMID- 11428443 TI - Outbreak of Serratia marcescens infection in a neonatal intensive care unit. AB - We report an outbreak of Serratia marcescens infection in the neonatal intensive care unit of a community hospital. The outbreak involved eight neonates, (five infected and three colonized), one of whom died. Pulsed-field gel electrophoresis confirmed that all isolates were identical strains. Cohorting and isolation of the infected neonates helped to control the outbreak. No environmental source of infection was found. PMID- 11428444 TI - The emerging nosocomial pathogens Cryptosporidium, Escherichia coli O157:H7, Helicobacter pylori, and hepatitis C: epidemiology, environmental survival, efficacy of disinfection, and control measures. AB - New and emerging infectious diseases pose a threat to public health and may be responsible for nosocomial outbreaks. Cryptosporidium parvum and Escherichia coli are gastrointestinal pathogens that have caused nosocomial infections via person to-person transmission, environmental contamination, or contaminated water or food. Helicobacter pylori has been transmitted via inadequately disinfected endoscopes. Finally, hepatitis C may be acquired by healthcare personnel by percutaneous or mucous membrane exposure to blood or between patients by use of contaminated blood products or via environmental contamination. Rigorous adherence to Standard Precautions, Contact Precautions for patients with infectious diarrhea, disinfection of environmental surfaces, and appropriate disinfection of endoscopes are adequate to prevent nosocomial acquisition of these pathogens. PMID- 11428445 TI - The chronic indwelling catheter and urinary infection in long-term-care facility residents. AB - From 5% to 10% of residents of long-term-care facilities have urinary drainage managed with chronic indwelling catheters. These residents are always bacteriuric, usually with a complex microbiological flora of two to five organisms and a biofilm on the catheter that may contribute to obstruction. Residents with chronic indwelling catheters have increased morbidity from urinary infection compared to bacteriuric residents without chronic catheters. The most effective means to prevent infection is limitation of chronic indwelling catheter use. While appropriate catheter care and infection control precautions are recommended in managing these patients, the impact of these practices on the occurrence of urinary infection or prevention of symptomatic episodes has not been evaluated. Symptomatic infection can likely be prevented by attention to catheter care, including early recognition and replacement of obstructed catheters and prevention of catheter trauma. Appropriate use of prophylactic antimicrobial therapy prior to invasive genitourinary procedures is also necessary. Asymptomatic bacteriuria should not be treated. When symptomatic episodes occur, patients should be evaluated clinically and microbiologically and treated with appropriate antimicrobial therapy. Further technological advances in catheter material and urine drainage will be needed to have a substantial impact on the frequency of urinary infection with chronic catheter use. PMID- 11428446 TI - CO2-laser treatment of recurrent glottic carcinoma. AB - BACKGROUND: Evaluation of the results of CO2 laser treatment of recurrent glottic carcinoma after radiotherapy. METHODS: Records of all patients treated in the University Hospital Rotterdam and the Dr. Daniel den Hoed Cancer Center between 1980 and 1996 by CO2 laser for recurrent glottic carcinoma were studied. RESULTS: Forty patients were treated by laser surgery. Average follow-up was 77 months. Twenty-three patients (58%) had another recurrence develop after laser surgery. Three were cured by a second laser procedure. Therefore, 20 patients (50%) were successfully treated with laser surgery. In 23 patients the recurrence did not extend into the anterior commissure; in 57% a total laryngectomy could be avoided. In 17 patients the recurrence did extend into the anterior commissure; in 41% a total laryngectomy was avoided. CONCLUSION: Most patients with recurrent carcinoma of the larynx after radiotherapy can be cured by laser surgery if the tumor does not extend into the anterior commissure. PMID- 11428447 TI - Outcome of patients treated with cobalt and 6 MV in head and neck cancers. AB - BACKGROUND: Since 1992, we have been using a 6-MV linear accelerator instead of a cobalt machine. The aim of the study is to evaluate the impact of this on neck control, particularly on postoperative patients in which subcutaneous tissues are at risk. METHOD: A retrospective study including all of 1,452 consecutive patients treated by definitive or postoperative radiotherapy between 1989 and 1997. All stages and subsites of the head/neck were included. Local and neck control were evaluated by the Kaplan Meier method, and comparisons were made between the cobalt and the 6-MV subgroups with a subsequent Cox analysis. For neck control analysis, the postoperative patients were divided in low and high risk (extracapsular extension [ECE], >two nodes, or T4). RESULTS: Radical radiotherapy: A better local control (LC) is observed with 6 MV than with cobalt, but neck control was similar. Postoperative radiotherapy: A better LC is observed with 6 MV. In high-risk patients, the neck control was higher for the cobalt group (79%) vs 60% for the 6-MV group (p = .09 and .03 in a Cox model). CONCLUSION: In postoperative patients at high risk for neck relapse, cobalt seems to give a better neck control. We are currently doing a prospective study in which a bolus is added for half the treatment when patients at high risk for neck relapse are treated with 6 MV. PMID- 11428448 TI - Selective use of drains in thyroid surgery. AB - OBJECTIVE: To identify whether there are differences in the use of drains and, if used, which would be the best for thyroid surgery. DESIGN: Prospective, longitudinal, comparative, randomized study. SETTING: General Hospital Mexico City, Mexico. PATIENTS AND METHODS: One hundred fifty patients were studied, divided into three groups: group A, without drain; group B, with a Penrose drain; and group C, with a semirigid suction drain. On the basis of the preoperative diagnosis, subtotal or total thyroidectomy or hemithyroidectomy was performed. Analyzed variables were thyroid volume (TV), transoperative bleeding (TOB), flow of postoperative drain (PD), length of hospital stay (HS), and complications, such as seromas, hematomas, and hemorrhages. Statistical Analysis. Multiple variant analysis, using Scheffe's procedure and chi2. RESULTS: Group A had an average TOB of 107 mL, HS of 2 days, and TV of 153.24 mL with two complications (seromas). Group B had an average TOB of 149.8 mL, HS of 2.6 days, TV of 175.4 mL, PD of 29.6 mL, and three complications (2 seromas and 1 hematoma). Group C had an average TOB of 161.5 mL, HS of 3.11 days, TV of 173.5 mL, PD of 25.84 mL, and two seromas. No differences existed regardless of the type of drain used between groups B and C. CONCLUSION: Statistical analysis showed that the size of the gland, diagnosis, type of surgery, transoperative bleeding, and complications are not valid arguments to leave an external drain in thyroid surgery. No advantages were found between the Penrose or the semirigid suction drains. Hospital stay was longer in patients with the suction drain. These results support the notion that the use of wound drainage cannot substitute for meticulous dissection and transoperative hemostasis. PMID- 11428449 TI - Presence of Epstein-Barr virus latent membrane protein 1 gene in the nasopharyngeal swabs from patients with nasopharyngeal carcinoma. AB - BACKGROUND: Nasopharyngeal carcinoma (NPC) is the most common head and neck malignancy in southeastern China and Taiwan. Early detection of the local disease followed immediately by proper treatment is essential to increase the cure and survival rates. Because every NPC tumor cell carries Epstein-Barr Virus (EBV) genomes, detection of EBV genomic DNA such as latent membrane protein 1 gene (LMP1) might indicate the presence of NPC. We developed a simple and noninvasive technique of nasopharyngeal swabbing to acquire nasopharyngeal cells for detecting the presence of EBV genome. The aim of this study was to investigate the feasibility and reliability of this technique. METHODS: We collected nasopharyngeal cells by means of a nasopharyngeal swabbing technique and detected the presence of EBV LMP1 with polymerase chain reaction (PCR). Thirty-eight swab specimens were obtained from patients with NPC who were newly diagnosed or were just beginning radiotherapy. Two groups of control subjects were recruited, including 20 patients with other head and neck cancers and eight family members of the NPC patients. An additional group of 65 NPC patients were enrolled in the course of regular follow-up after definitive radiotherapy. RESULTS: All of the samples yielded sufficient DNA for PCR amplification. Thirty-six of 38 NPC swab samples were positive for EBV LMP1, and all the control subjects had swab sample results negative for EBV. All five patients with suspected local recurrence exhibited positive EBV test results. CONCLUSIONS: Demonstration of EBV LMP1 in the nasopharyngeal swab specimens detected NPC with a sensitivity of 94.7% and specificity of 100%. This study confirms the reliability and feasibility of nasopharyngeal swab in the predicting and screening of NPC. PMID- 11428450 TI - A structured review of quality of life instruments for head and neck cancer patients. AB - BACKGROUND: Quality of life (QOL) is an important treatment outcome for head and neck cancer. Our purpose was to critically review published disease-specific QOL instruments. METHODS: Medline and Cancerlit were searched from 1966-1999. Eight disease-specific QOL instruments were identified, described, and appraised for development, sensibility, reliability, validity and responsiveness to change. RESULTS: Several of the available instruments have been well-developed and characterized. No one instrument is ideal for all purposes. When selecting a disease-specific QOL instrument for head and neck cancer patients, careful consideration must be given to disease subsite, treatment, timing of assessment, clinical setting, study purpose and research question. CONCLUSION: Validation of QOL instruments is an ongoing process. Direct comparisons of different instruments may help to establish the most appropriate questionnaire for each situation. Efforts should be focused on the evaluation of existing instruments, rather than the development of new questionnaires. PMID- 11428451 TI - How I do it: closure of tracheoesophageal puncture site. AB - BACKGROUND: Tracheoesophageal voice prosthesis is highly effective in providing speech after total laryngectomy. Although it is a safe method, tracheoesophageal fistulas occasionally need to be closed, usually at the request of the patient, because of leakage through an enlarging fistula. We present our technique for closure of tracheoesophageal fistula. METHODS: An incision is made at the mucocutaneous junction of the stoma from the 9 to the 3-o'clock position. Tracheoesophageal space is dissected down to and beyond the fistula. The tracheoesophageal tract is divided. The esophageal mucosa is closed with inverted sutures. After multiple layer closure of the esophageal fistula, the tracheal mucosa is closed with everted sutures. RESULTS: This technique has been used in nine patients. Eight were successful. The remaining patient had radiation therapy failure. CONCLUSION: This method of closure is simple and effective for those patients who require permanent closure of the tracheoesophageal fistula. PMID- 11428452 TI - Does type II nitric oxide synthase expression correlate with cellular proliferation in oral squamous cell carcinoma and dysplasia? AB - BACKGROUND: Nitric oxide (NO) has been implicated both in tumor progression and inhibition. This study investigated whether type II nitric oxide synthase (NOS2) expression correlated with cell proliferation in oral squamous cell carcinoma (OSCC) and dysplasia. METHODS: Paraffin-embedded tissue samples of normal oral mucosa, OSCC, and dysplasia were assessed immunohistochemically using monoclonal antibodies to NOS2 and Ki-67 antigen. We used Western blotting to confirm NOS2 antibody specificity and protein expression in select cases. RESULTS: NOS2 staining was increased in OSCC relative to normal oral mucosa, in which no expression was found. Both NOS2 expression and Ki-67 indices independently correlated with grade of dysplasia (p < .001) but not with the degree of tumor differentiation. A positive correlation was found between NOS2 expression and Ki 67 in cases of mild and moderate dysplasia (p < .001), but not in severe dysplasia and OSCC. CONCLUSIONS: No correlation exists between Ki-67 and NOS2 expression in severe dysplasia and OSCC. The findings suggest that the level of NO produced by NOS2 is insufficient to affect cellular proliferation in these conditions. The mechanism of NOS2 activation and the consequences of its expression remain to be fully explained. PMID- 11428453 TI - Growth inhibition and chemosensitivity of poorly differentiated human thyroid cancer cell line (NPA) transfected with p53 gene. AB - BACKGROUND: We investigated whether retroviral p53 transfection could enhance growth inhibition and chemosensitivity in a p53 mutant papillary thyroid cancer cell line (NPA). METHODS: NPA cells were transfected with either LXSN/p53 or mock infection in the presence of Adriamycin. Gene expression was confirmed by western blotting. Nude mice were injected subcutaneously with NPA cells after transfection with either LXSN/p53 or mock infection on opposite sides, and the tumor growth was compared. RESULTS: There was a dose-dependent inhibition of tumor growth with LXSN/p53 transfection. Tumor growth was inhibited more by p53 gene transfection relative to mock transfection in the presence of Adriamycin. CONCLUSION: These treatment modalities could be beneficial in the treatment of p53 mutant positive thyroid cancers. PMID- 11428454 TI - Relocating the site of frozen sections--is there room for improvement? AB - INTRODUCTION: In an attempt to improve the marginal control of oropharyngeal carcinoma, some surgeons routinely perform frozen section analysis. Because current methods of relocating the site from which frozen section specimens are harvested can be haphazard, we studied the accuracy of a common technique used to localize specimens to the resected tumour bed. METHODS: One surgeon was asked to identify the sites of proposed sampling in 14 consecutive cases. After approximately 5 minutes, the same surgeon was asked to relocate each site. RESULTS: In all, 71 soft tissue points were identified. The mean error in relocating the sample site was 9 mm for those placed at mucosal margins and 12 mm for those placed deep to the tumor bed. The error exceeded 1 cm in 32% (23 of 71) of cases. CONCLUSION: These findings highlight the need to accurately locate the position of frozen sections if samples that subsequently prove positive are to used to greatest effect. PMID- 11428455 TI - Intraoperative B-mode endosonography of tongue carcinoma. AB - BACKGROUND: Exact estimation of a tumor's size and definition of adequate resection margins in carcinomas of the tongue are often difficult because of the tumor's extension and deep infiltration. METHODS: We developed a method that allows intraoperative visualization and marking of tumor margins: intraoperative endosonography was performed in five patients with carcinomas of the tongue with an 8- to 12-MHz linear array transducer. The oral cavity was flooded with normal saline solution, and the transducer was immersed therein. This allowed scanning in a noncontact mode. The tumor margins were marked with a surgical suture under endosonographic monitoring. RESULTS: In the five patients studied, histologic margins corresponded to sonographic margins. The sonographic marking proved to be useful during the resection of the tumor, and histologic safety margins were respected in each case. CONCLUSIONS: This noninvasive procedure provides a quick and reliable orientation during resection of tongue carcinoma, and a more precise and individual definition of resection margins is possible. Intraoperative noncontact use of endosonography is a promising method, and further studies may confirm this. PMID- 11428456 TI - Chromosomal alterations in squamous cell carcinomas of the head and neck: window to the biology of disease. AB - BACKGROUND: Cytogenetic alterations underlie the development of squamous cell carcinomas of the head and neck (SCCHN). Because many of the molecular genetic changes in SCCHN result from chromosomal alterations, a complete perspective on the genetic changes in tumors requires a basic introduction to cytogenetics. This review presents a brief description of the latest cytogenetic techniques and a description of chromosomal alterations in SCCHN, their molecular correlates, and clinical implications. RESULTS: The most frequent cytogenetic alterations in SCCHN are gains of 3q, 8q, 9q, 20q, 7p, 11q13, and 5p and losses of 3p, 9p, 21q, 5q, 13q, 18q, and 8p. The karyotypes often provide an explanation for the mechanism by which the molecular genetic alterations arose. For example, the coordinate gains and losses involving whole arms of chromosomes 3, 5, 7, 8, and 9 often result from isochromosome formation. In addition, apparent allelic imbalances may not represent loss of heterozygosity but gene amplification. These results suggest that cytogenetic analysis is valuable for placing the molecular genetic findings in perspective at the cellular level. CONCLUSIONS: Cytogenetic endpoints may be useful tools for dissecting clinical differences in tumor behavior and response to therapy. Numerous studies are underway to examine the biology of and genetic alterations in SCCHN that will lead to additional markers for use as rapid, noninvasive screening methods for individuals at high risk for primary or recurrent SCCHN. Our goal is to minimize morbidity and mortality from SCCHN by identifying useful predictors of disease and recurrence risk and response to therapy to implement earlier detection and more effective prevention and/or treatment strategies. PMID- 11428457 TI - MALT lymphoma presenting as a cystic salivary gland mass. AB - BACKGROUND: Mucosa associated lymphoid tissue (MALT) lymphoma has been noted to involve the salivary glands in chronic inflammatory conditions such as Sjogren's syndrome and in HIV infection. METHODS AND RESULTS: The authors encountered a patient with bilateral cystic changes in the parotid glands which proved to be due to MALT lymphoma. The clinical course, histopathology, and treatment options of MALT lymphoma in the salivary gland are discussed in detail. CONCLUSION: This malignant entity should be considered in the differential diagnosis of refractory cystic lesions of the salivary glands. PMID- 11428458 TI - Pseudoaneurysm of the proximal facial artery presenting as oropharyngeal hemorrhage. AB - BACKGROUND: Penetrating trauma to the neck traversing zones II and III may cause considerable damage to soft tissues and neurovascular structures. Delayed sequelae of vascular injuries, such as pseudoaneurysm (PA), may present weeks to months after the initial injury. METHODS: We report an unusual case of a traumatic PA of the proximal facial artery that ruptured into the oropharynx. RESULTS: A 30-year-old man presented with oropharyngeal hemorrhage one month after a gunshot wound to the neck. Angiography revealed a PA of the proximal facial artery, which was treated with embolization. The arterial injury leading to the pseudoaneurysm had not been detected by arteriography at the time. CONCLUSIONS: PAs are rare complications of penetrating neck trauma. To our knowledge, this is only the second report of PA involving the proximal facial artery, and the first of a facial PA rupture into the pharynx. PMID- 11428459 TI - Ikirara insertions reveal five new Anopheles gambiae transposable elements in islands of repetitious sequence. AB - Characterization of Anopheles gambiae genomic clones containing Ikirara inverted repeats revealed five novel sequences related to known transposable elements (TEs). One TE is related to the mariner/Tc1 superfamily of class II (DNA-to-DNA) transposons, while four are related to class I (RNA-mediated transposition) elements. Crusoe, the class II element; is most similar to the Caenorhabditis elegans transposon Tc1-like TEs. Vash elements, represented twice in our clones, are related to the Q/T1 family of A. gambiae non-LTR retrotransposable elements. Guildenstern is a member of the RT1 and RT2 non-LTR retrotransposon family. Although RT1 and RT2 elements normally have a highly stereotyped insertion preference for sequences within ribosomal genes, Guildenstern is not located in ribosomal sequence. JuanAg is the first anopheline member of the mosquito non-LTR retrotransposon family of Juan elements that previously had included just the culicine elements JuanA and JuanC. Approximately 753 bp is missing from the central portion of the JuanAg reverse transcriptase gene, where an Ikirara inverted repeat is found in its stead. Ozymandias, the only LTR retrotransposon found in the clones, is most similar to the Drosophila melanogaster 412 element. Single Ikirara inverted repeats were also found adjacent to nontransposable element repetitious sequences. Our analysis suggests that the A. gambiae genome organization could best be described as islands of short-period interspersion repetitious DNA in a sea of long-period interspersion, mostly unique sequence DNA. PMID- 11428460 TI - Phylogenetic analysis of the Friedreich ataxia GAA trinucleotide repeat. AB - Friedreich ataxia is an autosomal recessive neurodegenerative disorder associated with a GAA repeat expansion in the first intron of the gene (FRDA) encoding a novel, highly conserved, 210 amino acid protein known as frataxin. Normal variation in repeat size was determined by analysis of more than 600 DNA samples from seven human populations. This analysis showed that the most frequent allele had nine GAA repeats, and no alleles with fewer than five GAA repeats were found. The European and Syrian populations had the highest percentage of alleles with 10 or more GAA repeats, while the Papua New Guinea population did not have any alleles carrying more than 10 GAA repeats. The distributions of repeat sizes in the European, Syrian, and African American populations were significantly different from those in the Asian and Papua New Guinea populations (p < 0.001). The GAA repeat size was also determined in five nonhuman primates. Samples from 10 chimpanzees, 3 orangutans, 1 gorilla, 1 rhesus macaque, 1 mangabey, and 1 tamarin were analyzed. Among those primates belonging to the Pongidae family, the chimpanzees were found to carry three or four GAA repeats, the orangutans had four or five GAA repeats, and the gorilla carried three GAA repeats. In primates belonging to the Cercopithecidae family, three GAA repeats were found in the mangabey and two in the rhesus macaque. However, an AluY subfamily member inserted in the poly(A) tract preceding the GAA repeat region in the rhesus macaque, making the amplified sequence approximately 300 bp longer. The GAA repeat was also found in the tamarin, suggesting that it arose at least 40 million years ago and remained relatively small throughout the majority of primate evolution, with a punctuated expansion in the human genome. PMID- 11428461 TI - Evolution of the aldose reductase-related gecko eye lens protein rhoB-crystallin: a sheep in wolf's clothing. AB - rhoB-crystallin (AJ245805) is a major protein component (20%) in the eye lens of the gecko Lepidodactylus lugubris. Limited peptide sequence analysis earlier revealed that it belongs to the aldo-keto reductase superfamily, as does the frog lens rho-crystallin. We have now determined the complete cDNA sequence of rhoB crystallin and established that it is more closely related to the aldose reductase branch of the superfamily than to frog rho-crystallin. These gecko and frog lens proteins have thus independently been recruited from the same enzyme superfamily. Aldose reductase is implicated in the development of diabetic cataract in mammals, and, if active, rhoB-crystallin might be a potential risk for the gecko lens. Apart from a replacement 298 Cys --> Tyr, rhoB-crystallin possesses all amino acid residues thought to be required for catalytic activity of the aldose reductases. However, modeling studies of the rhoB-crystallin structure indicate that substrate specificity and nicotinamide cofactor affinity might be affected. Indeed, neither recombinant rhoB-crystallin nor the reverse mutant 298 Tyr --> Cys showed noticeable activity toward aliphatic and aromatic substrates, although cofactor binding was retained. Various other oxidoreductases are known to be recruited as abundant lens proteins in many vertebrate species; rhoB-crystallin demonstrates that an aldose reductase-related enzyme also can be modified to this end. PMID- 11428462 TI - The comparative genomics of polyglutamine repeats: extreme differences in the codon organization of repeat-encoding regions between mammals and Drosophila. AB - Polyglutamine repeats within proteins are common in eukaryotes and are associated with neurological diseases in humans. Many are encoded by tandem repeats of the codon CAG that are likely to mutate primarily by replication slippage. However, a recent study in the yeast Saccharomyces cerevisiae has indicated that many others are encoded by mixtures of CAG and CAA which are less likely to undergo slippage. Here we attempt to estimate the proportions of polyglutamine repeats encoded by slippage-prone structures in species currently the subject of genome sequencing projects. We find a general excess over random expectation of polyglutamine repeats encoded by tandem repeats of codons. We nevertheless find many repeats encoded by nontandem codon structures. Mammals and Drosophila display extreme opposite patterns. Drosophila contains many proteins with polyglutamine tracts but these are generally encoded by interrupted structures. These structures may have been selected to be resistant to slippage. In contrast, mammals (humans and mice) have a high proportion of proteins in which repeats are encoded by tandem codon structures. In humans, these include most of the triplet expansion disease genes. PMID- 11428463 TI - Vertebrate LTR retrotransposons of the Tf1/sushi group. AB - LTR retrotransposons of the Tf1/sushi group from a diversity of vertebrates, including fish, amphibians, and mammals (humans, mice, and others), are described as full-length or partial elements. These elements are compared, and the mechanisms involved in self-priming of reverse transcriptase and programmed phase shifting are inferred. Evidence is presented that in mammals these elements are still transcriptionally active and are represented as proteins. This suggests that members of the Tf1/sushi group are present as functional elements (or incorporated as partial elements into host genes) in diverse vertebrate lineages. PMID- 11428464 TI - Synonymous codon usage, accuracy of translation, and gene length in Caenorhabditis elegans. AB - In many unicellular organisms, invertebrates, and plants, synonymous codon usage biases result from a coadaptation between codon usage and tRNAs abundance to optimize the efficiency of protein synthesis. However, it remains unclear whether natural selection acts at the level of the speed or the accuracy of mRNAs translation. Here we show that codon usage can improve the fidelity of protein synthesis in multicellular species. As predicted by the model of selection for translational accuracy, we find that the frequency of codons optimal for translation is significantly higher at codons encoding for conserved amino acids than at codons encoding for nonconserved amino acids in 548 genes compared between Caenorhabditis elegans and Homo sapiens. Although this model predicts that codon bias correlates positively with gene length, a negative correlation between codon bias and gene length has been observed in eukaryotes. This suggests that selection for fidelity of protein synthesis is not the main factor responsible for codon biases. The relationship between codon bias and gene length remains unexplained. Exploring the differences in gene expression process in eukaryotes and prokaryotes should provide new insights to understand this key question of codon usage. PMID- 11428465 TI - Ancient mitochondrial DNA reveals the origin of Sus scrofa from Rebun Island, Japan. AB - The Kabukai A site (5 to 8C A.D.) of the Okhotsk cultural area is on Rebun Island, a small island near the coast, north-northwest of Hokkaido, Japan. Specimens of Sus scrofa, called the Sakhalin pig, were discovered in five cultural layers at the Kabukai A site. Ancient DNA was extracted from the remains of 42 Sakhalin pig bones. Thirty-nine nucleotide sequences of the 574-bp mitochondrial DNA control region, estimated to have originated from at least 21 individuals, were amplified and analyzed phylogenetically. Nine distinct haplotypes (A1, A2, A3, B1, B2, C1, C2, D1, and D2) from this site were classified into four haplotype groups (A, B, C, and D) by parsimonious network analysis. Phylogenetic analysis of 9 ancient and 55 modern haplotypes indicated that the population of Sakhalin pigs at the Kabukai A site belonged to two distinct clusters; haplotype groups A and B formed a cluster comprised only of themselves, and haplotype groups C and D belonged to the cluster of one of the two genetic groups of Japanese wild boars uniquely distributed in the western part of Japan, including one northeast Mongolian wild boar. Analysis of the haplotype distribution among three archaeological sites and their historical transitions among the five layers reflecting the cultural periods at the Kabukai A site suggests that the Sakhalin pig populations were introduced from Sakhalin island and the Amur River basin in the northeastern Eurasian continent together with some cultural influences. PMID- 11428466 TI - Evidence for multiple origins of human infectivity in Trypanosoma brucei revealed by minisatellite variant repeat mapping. AB - In recent years a wide variety of biochemical and molecular typing systems has been employed in the study of parasite diversity aimed at investigating the level of genetic diversity and delineating the relationship between different species and subspecies. However, such methods have failed to differentiate between two of the classically defined subspecies of the protozoan parasite Trypanosoma brucei: the human infective, T. b. rhodesiense, which causes African sleeping sickness, and the non-human infective T. b. brucei. This has led to the hypothesis that T. b. rhodesiense is a host range variant of T. b. brucei. In this paper we test this hypothesis by examining highly polymorphic tandemly repeated regions of the trypanosome genome, i.e., minisatellite loci. We have employed the technique of minisatellite variant repeat mapping by PCR (MVR-PCR), which determines the distribution of variant repeat units along the tandem array of one minisatellite, MS42. The maps generated by this technique not only allow unequivocal allele identification but also contain within them cladistic information which we used to determine the possible genetic relationship between the different subspecies of T. brucei. Our findings revealed that human infective (T. b. rhodesiense) isolates from Uganda are more closely related to the local non-human infective isolates (T. b. brucei) than they are to other human infective stocks from different regions, suggesting that human infectivity has originated independently in these different geographical regions. This would infer that the separate classification of all human infective stocks from East Africa into the subspecies T. b. rhodesiense is genetically inappropriate and it would be better to consider geographically separate populations as host range variants of T. brucei brucei or perhaps as a series of different subspecies. Based on these data, it is clear that MVR mapping is a very useful tool for the analysis of zoonotic eukaryotic pathogens where delineation of the origins of outbreaks of disease and definition of human infective strains are key questions. PMID- 11428467 TI - A phylogenetic study of the origin of the domestic pig estimated from the near complete mtDNA genome. AB - The near-complete pig mtDNA genome sequence (15,997 bp) was determined from two domestic pigs (one Chinese Meishan and one Swedish Landrace) and two European wild boars. The sequences were analyzed together with a previously published sequence representing a Swedish domestic pig. The sequences formed three distinct clades, denoted A, E1, and E2, with considerable sequence divergence between them (0.8-1.2%). The results confirm our previous study (based on the sequence of the cytochrome B gene and the control region only) and provide compelling evidence that domestication of pigs must have occurred from both an Asian and a European subspecies of the wild boar. We estimated the time since the divergence of clade A (found in Chinese Meishan pigs) and E1 (found in European domestic pigs) at about 900,000 years before present, long before domestication about 9,000 years ago. The pattern of nucleotide substitutions among the sequences was in good agreement with previous interspecific comparisons of mammalian mtDNA; the lowest substitution rates were observed at nonsynonymous sites in protein-coding genes, in the tRNA and rRNA genes, while the highest rates were observed at synonymous sites and in the control region. The presence of Asian clade A in some major European breeds (Large White and Landrace) most likely reflects the documented introgression of Asian germplasm into European stocks during the 18th and 19th centuries. The coexistence of such divergent mtDNA haplotypes for 100+ generations is expected to lead to the presence of recombinant haplotypes if paternal transmission and recombination occur at a low frequency. We found no evidence of such recombination events in the limited sample studied so far. PMID- 11428468 TI - Management of coronary artery disease in patients with diabetes mellitus. PMID- 11428469 TI - Percutaneous coronary intervention in patients with unstable angina. PMID- 11428470 TI - Failed thrombolysis: a continuing problem. PMID- 11428471 TI - A randomized study of the safety and efficacy of reused angioplasty balloon catheters. AB - BACKGROUND: To lower costs, many centers around the world utilize previously used, resterilized balloon catheters to perform coronary angioplasty There are no controlled trials regarding their safety and efficacy. METHODS AND RESULTS: We performed the first randomized, double-blind, controlled, single-center clinical trial comparing the safety (clinical success) and efficacy (angiographic success) of reused versus new coronary angioplasty balloon catheters. A total of 377 procedures were included, 178 in the reused catheter arm and 199 in the new catheter arm. There were no significant differences in clinical or lesion characteristics among the two arms. The incidence of first balloon failure in the reused catheter arm was similar to that of the new catheter arm (12 cases [7%] v. 10 cases [5%], respectively). The angiographic success rate was also similar-176 cases (98.9%) in the reused catheter arm and 196 cases (98.5%) in the new catheter arm. The number of balloon catheters used per lesion, amount of contrast, and procedural and fluoroscopy time were similar in the two arms. At 30 days, the incidence of major adverse cardiac events was similar in both arms, 8 cases (4.5%) in the reused catheter arm and 10 cases (5%) in the new catheter arm. The incidence of fever was also similar. CONCLUSIONS: When performing coronary angioplasty, reused catheters are as effective (similar angiographic success) and safe (similar clinical success) as new catheters. PMID- 11428472 TI - Lipid-lowering effect of simvastatin in patients of type 2 diabetes mellitus. AB - BACKGROUND: Dyslipidemia is an important factor in causation of macrovascular disease in type 2 diabetics. The role of simvastatin in the management of dyslipidemia in patients with type 2 diabetes mellitus is not very well elucidated, particularly in the context of the recent American Diabetes Association criteria 2001. The American Diabetes Association suggests that aggressive therapy of diabetic dyslipidemia will reduce the risk of coronary heart disease in diabetics and that optimal levels are serum low-density lipoprotein cholesterol <2.60 mmol/L (< 100 mg/dl), high-density lipoprotein cholesterol >1.1 5 mmol/L (>45 mg/dl) and triglycerides <2.30 mmol/L, (<200 mg/dl). This study was planned to compare the effect of simvastatin together with behavioral modification and behavioral modification alone, in age, sex and body mass index matched patients with type 2 diabetes mellitus with dyslipidemia, in reaching the target levels of various lipids as suggested by the American Diabetes Association criteria 2001. METHODS AND RESULTS: An open-label, prospective study was conducted on 80 patients with type 2 diabetes mellitus, who had fair to moderate glycemic control with a total glycated hemoglobin < 10%. The patients in the control group (n=40) were treated with only behavioral modifications like calorie control and daily walking for 30 minutes, and no lipid lowering agent was given. The lipid profile was re-evaluated after 6 and 12 weeks. The patients in the test group (n=40) were advised behavioral modification and given simvastatin. The starting dose was 10 mg at bed time. After 6 weeks of simvastatin therapy, a lipid profile was done. If the goal of low-density lipoprotein cholesterol < 100 mg/dl and/or triglycerides <200 mg/dl and/or high density lipoprotein cholesterol >45 mg/dl was not achieved, the dose of simvastatin was increased to 20 mg at bedtime for another 6 weeks. It was observed that low-density lipoprotein dyslipidemia was most prevalent. In the control group, a favorable alteration in lipid levels was brought about but none was statistically significant and the American Diabetes Association goals were not achieved in any of the patients. In the test group, there was a significant and favorable alteration in all lipid moieties, and the target levels were achieved in 80% of patients after 12 weeks. There was no significant alteration in glycemic control and liver functions. Myopathy and epigastric pain were seen in 1 patient in each group. CONCLUSIONS: In our study, behavioral modification alone did not achieve the target levels of various lipids in diabetic dyslipidemia as per the American Diabetes Association guidelines. Hence, pharmacological therapy with statins should be resorted to in patients with type 2 diabetes mellitus who carry a high risk of coronary heart disease. Simvastatin is a safe and efficacious lipid-lowering drug. PMID- 11428473 TI - Relationship of Xba1 and EcoR1 polymorphisms of apolipoprotein-B gene to dyslipidemia and obesity in Asian Indians in North India. AB - BACKGROUND: Genetic investigation of dyslipidemia and obesity prevalent in the Indian population form the basis of this study. METHODS AND RESULTS: The frequency of restriction fragment length polymorphisms (Xba1 and EcoR1) of the apolipoprotein-B gene was investigated in a case-control study of 30 hyperlipidemic and 40 normolipidemic subjects. By univariate analysis, old age, higher body mass index, waist-hip ratio and sum of four skinfolds were found to be significantly associated with hyperlipidemia. The frequencies of X- and E+ alleles of the apolipoprotein-B gene were significantly higher in North Indians in the state of New Delhi (0.83 and 0.91, respectively) as compared to the observations made in Caucasians in previous studies, but was similar to the frequency reported in Indians settled in Singapore and the UK. There were no significant differences in the allele or genotype frequencies of either Xba1 or EcoR1 polymorphisms between the hyperlipidemic and normolipidemic groups. On multiple logistic regression analysis considering body mass index, waist-hip ratio, percentage body fat and genotypes as independent variables, no association was observed between the apolipoprotein-B genotypes and serum lipid components. Further, there were no associations between apolipoprotein-B polymorphisms and generalized obesity (as assessed by body mass index, sum of four skinfolds, and percentage total body fat) and abdominal obesity (as measured by waist circumference and waist-hip ratio). CONCLUSIONS: We conclude that apolipoprotein B (Xba1 and EcoR1) polymorphisms do not appear to influence serum lipid levels and parameters of generalized andregional obesity in the study sample. PMID- 11428474 TI - The isoprenoid pathway in lone atrial fibrillation with embolic stroke. AB - BACKGROUND: The isoprenoid pathway was assessed and compared in patients of lone atrial fibrillation with embolic stroke as well as in patients with right hemispheric, left hemispheric and bihemispheric dominance to determine the role of hemispheric dominance in its pathogenesis. METHODS AND RESULTS: The activities of hydroxyl methyl glutaryl-CoA reductase and RBC sodium-potasium ATPase as well as serum levels of plasma magnesium, digoxin, dolichol and ubiquinone were measured. The tyrosine/tryptophan catabolic patterns, glycoconjugate metabolism, free radical metabolism and RBC membrane composition were also assessed. In patients with lone atrial fibrillation with embolic stroke, there was elevated digoxin synthesis, increased dolichol and glycoconjugate levels, and low ubiquinone and elevated free radical levels. There was also an increase in tryptophan catabolites and a reduction in tyrosine catabolites: and an increase in the cholesterol: phospholipid ratio with a reduction in the glycoconjugate levels of the RBC membrane. The same biochemical patterns were obtained in individuals with right hemispheric dominance whereas the patterns were reversed in patients with left hemispheric dominance. CONCLUSIONS: Lone atrial fibrillation with embolic stroke is associated with an upregulated isoprenoid pathway and elevated digoxin secretion from the hypothalamus. This occurs in right hemisphere-dominant individuals. PMID- 11428475 TI - Prevalence of coronary artery disease in patients with symptomatic peripheral vascular disease. AB - BACKGROUND: Prevalence of coronary artery disease has been reported to be quite high in patients with peripheral vascular disease in western literature. Therefore, it is important to study the coronary anatomy in patients with symptomatic peripheral vascular disease. METHODS AND RESULTS: Fifty-three patients presenting with symptoms of peripheral vascular disease underwent peripheral angiography in our institute during the last 2 years. The total number of vessels involved in these patients was 117. Fifteen patients had involvement of the upper limb vessels, 46 patients had involvement of the lower limb vessels and 6 patients had involvement of the carotid/vertebral arteries. Coronary arteriography was done in all the patients. Only 8 (15%) patients were found to have coronary artery disease with involvement of 11 arteries. Eighty-four (72%) peripheral vessels out of the 117 vessels involved showed total occlusion, whereas only 2 (18%) coronary arteries out of 11 vessels involved showed total occlusion. CONCLUSIONS: This study shows that the majority of patients with symptomatic peripheral vascular disease have normal coronaries, the extent of their involvement being low despite severe peripheral vascular disease. PMID- 11428476 TI - Percutaneous transseptal mitral commissurotomy in pregnant women with critical mitral stenosis. AB - BACKGROUND: Percutaneous transseptal mitral commissurotomy has been successfully performed in selected pregnant patients with severe symptomatic mitral stenosis. Its safety and efficacy needs to be evaluated in a large number of cases. METHODS AND RESULTS: Percutaneous transseptal mitral commissurotomy was performed in 85 severely symptomatic (New York Heart Association functional class III or IV) pregnant women aged 22.7+/-4.1 years (range 18-39 years) with critical mitral stenosis at 24.8+/-4.7 weeks (range 20-34 weeks) of gestation. Percutaneous valvotomy was performed using a flow-guided Inoue balloon in all the patients. The procedure was considered successful in 80 (94%) patients. The hemodynamic mean end-diastolic gradient decreased from 26.7+/-6.8 mm Hg (range 16-35 mmHg) to 4.5+/-3.8 mmHg (range 0-14 mmHg) (p<0.001). The mean diastolic gradient decreased from 29.1+/-9.1 mmHg (range 18-38 mmHg) to 7.2+/-4.1 mmHg (range 4.1-18 mmHg) (p<0.001). The mean mitral valve area assessed by echocardiography increased from 0.75+/-0.5 cm2 (range 0.4-1.0 cm2) to 2.0+/-0.5 (range 1.0-2.7 cm2) (p<0.001). The mean fluoroscopy time was 3.6+/-3.2 minutes. The results of the mitral valvotomy were considered suboptimal in 4 patients. Mitral regurgitation increased by 1 grade in 16 patients and more than 2 grades in 2 patients. One patient developed pericardial tamponade during the procedure and was managed by catheter drainage. Percutaneous mitral valve dilatation was then successfully performed in this patient. No fetal abortion occurred after the procedure. CONCLUSIONS: The results of this study indicate that percutaneous transseptal mitral commissurotomy is a safe and effective procedure for severe symptomatic mitral stenosis in pregnancy. PMID- 11428477 TI - A meta-analysis of controlled clinical trials comparing low-molecular weight heparins with unfractionated heparin in unstable angina. AB - BACKGROUND: Unfractionated heparin has been used extensively for the treatment of unstable angina/non-Q wave myocardial infarction but it has several disadvantages. Low-molecular weight heparins are now recommended although they are 3-5 times costlier than unfractionated heparin since they are convinient to administer and do not require activated thromboplastin time monitoring. Whereas enoxaparin, a low-molecular weight heparin, has been demonstrated to be superior to unfractionated heparin, the results of other low-molecular weight heparins have not been so convincing. METHOD AND RESULTS: Through manual, MEDLINE and EMBASE search, we identified five randomized trials (excluding enoxaparin trials) that compared low-molecular weight heparins with unfractionated heparin in unstable angina. The prespecified efficacy end point of interest included a composite of death, myocardial infarction, recurrent angina and urgent revascularization. The safety end point was taken as a composite of major hemorrhage, minor hemorrhage, thrombocytopenia, allergic reaction and any other adverse event. We calculated odds ratio (95% confidence interval) for each trial for the composite end point, and the pooled odds ratio (95%) confidence interval) was calculated using two established methods of meta-analysis, the Mantel Haenszel-Peto method and the DerSirmonian-Laird method. Both the methods yielded similar odds ratio (95% confidence interval). Separate odds ratio were calculated for efficacy and safety end points. There was a nonsignificant reduction in the incidence of the composite efficacy end point: the odds ratio (95% confidence interval) was 0.83 (0.70-0.99: p=0.08). The odds ratio (95% confidence interval) for the safety data was 0.78 (0.69-1.26: p=0.33). CONCLUSIONS: No statistically significant difference was observed when the efficacy and safety of low-molecular weight heparins were compared with those of unfractionated heparin. A cost effectiveness analysis of low-molecular weight heparins versus unfractionated heparin must be done urgently to establish more firmly the place of low-molecular weight heparins in the management of unstable angina. PMID- 11428478 TI - Brugada syndrome with monomorphic ventricular tachycardia in a one-year-old child. AB - A one-year-old child with a structurally normal heart presented with monomorphic ventricular tachycardia. Electrocardiogram in sinus rhythm showed right bundle branch block with ST segment elevation suggesting a diagnosis of Brugada syndrome. At a later date, when the ST segment was isoelectric. intravenous procainamide caused ST elevation typical of Brugada syndrome. PMID- 11428479 TI - Valvuloplasty balloon detaching from the stem of a catheter. AB - A 12-year-old boy underwent pulmonary balloon valvotomy for isolated critical pulmonary stenosis. Following valvotomy, blood was found to be drawn into the syringe during deflation of the balloon, so a provisional diagnosis of a burst balloon was made. However, when the balloon catheter was withdrawn, the balloon got detached from the stem of the catheter at the level of the right atrium and was retained over the exchange guidewire. The balloon, when retrieved with a snare, was found to be intact. The balloon may have been partially detached at the junction of the proximal end of the balloon and the catheter; hence, blood was drawn from the catheter during deflation. In our institution balloons are reused following sterilization with ethylene oxide gas. We conclude that any balloon presumed to have burst inside the heart must be removed with great caution. In a third world country like India, where cost is an important factor, balloons can be reused, but with caution, keeping in mind complications such as in this case. PMID- 11428480 TI - Adenosine-induced ventricular fibrillation. AB - The use of adenosine has been suggested as a diagnostic tool in the evaluation of wide ORS complex tachycardia. However, adenosine shortens the antegrade refractoriness of accessory atrioventricular connections and may cause acceleration of the ventricular rate during atrial fibrillation. We observed ventricular fibrillation in 2 patients who presented to the emergency department with pre-excited atrial fibrillation and were given 12 mg of adenosine. PMID- 11428481 TI - Isolated congenital left ventricular diverticulum in adults. AB - Isolated congenital ventricular diverticulum or aneurysm is rare and usually arises from the left ventricle. The presentation of this condition is diverse. We report three cases of isolated congenital left ventricular diverticula. The age range was 17-30 years. Chest X-ray provided the earliest clinical suspicion in these three cases of a cardiac anomaly which was diagnosed by echocardiography and confirmed by angiocardiography. The location of the congenital left ventricular diverticulum was the left ventricular apex in two cases and basal in the other. We conclude that congenital left ventricular diverticulum is a disease of protean presentations. A high index of suspicion is necessary while interpreting chest X-rays and echocardiographs to diagnose congenital left ventricular diverticulum. A contractile accessory chamber of the left ventricle with a narrow neck with or without midline defects and an electrocardiogram without Q waves is consistent with the diagnosis of congenital left ventricular diverticulum. PMID- 11428482 TI - Stenting for SVC obstruction in an infant operated for total anomalous pulmonary venous return. AB - Superior vena cava obstruction following corrective repair of total anomalous pulmonary venous return has rarely been described in the literature. A one-month old boy who underwent corrective surgery for obstructive supracardiac total anomalous pulmonary venous return with consequent symptomatic superior vena cava obstruction in the immediate postoperative period, is reported. This was treated by balloon dilatation followed by stenting of the superior vena cava. The immediate postoperative result was satisfactory and the infant continued to remain asymptomatic at six months follow up. We suggest that this intervention could prove to be a viable alternative to a repeat surgical procedure for such complex cases. PMID- 11428483 TI - Pulmonary embolism with isolated right ventricular infarction. AB - Concomitant occurrence of pulmonary embolism and right ventricular infarction is rare. It poses important diagnostic and therapeutic implications. A case of pulmonary embolism with isolated right ventricular anterior wall infarction presented with ventricular tachycardia. One pathology could have led to the other. Two-dimensional echocardiography was useful in documenting pulmonary artery hypertension as well as regional wall motion abnormality of the right ventricle. Thrombolytic therapy and dobutamine infusion were useful. Nitrates, fluid infusion and diuretics should be used cautiously. PMID- 11428484 TI - Secondary lymphoma of the heart presenting as recurrent syncope. AB - A 12-year-old male child presented with recurrent syncope. Ventricular tachycardia was noted on the electrocardiogram. Transthoracic echocardiogram revealed a homogeneous tumor mass in the right ventricular cavity with extension into the outflow region. Left cervical lymph node biopsy confirmed the diagnosis of non-Hodgkin's lymphoma. The tumor resolved completely with chemotherapy without surgical intervention. PMID- 11428485 TI - Does moderate alcohol intake protect against coronary heart disease? AB - There is abundant epidemiological and clinical evidence to show that light-to moderate drinking is associated with a reduced risk of coronary heart disease (CHD), total and ischemic stroke, and total mortality in middle-aged and elderly men and women. The evidence suggests a J- or U-shaped relationship between alcohol and CHD. Alcohol reduces the risk of coronary heart disease both by inhibiting the formation of atheroma and by decreasing the rate of blood coagulation. It appears that for most conditions, other than cardiovascular diseases and cholelithiasis, moderate alcohol consumption has either none or only an intermediate type of risk as compared with the risk of either abstinence or excessive drinking. It is now fully recognized and accepted that drinking alcohol regularly for years is toxic to almost every tissue of the body. However, most people who choose to drink alcohol have little or no problem limiting their consumption to amounts that do not generally cause serious health or social consequences. Moreover, a given dose of alcohol may affect different people differently. It is, therefore, imperative that a critical evaluation, based on the observations made hitherto, be done of both the harmful and the protective effects of alcohol consumption on various organs/systems of the body. This article reviews epidemiological evidence for the protective effects of alcohol on the cardiovascular system and discusses how alcohol might lower the risk of CHD. PMID- 11428486 TI - Communications in cardiology: why digital? PMID- 11428487 TI - Pulmonary vein varix in association with bilateral pulmonary vein stenosis. PMID- 11428488 TI - Effects of atorvastatin on early recurrent ischemic events in acute coronary syndromes. The MIRACL study. PMID- 11428489 TI - Unprotected left main coronary artery stenting: immediate and medium-term outcomes of 140 elective procedures. PMID- 11428490 TI - Arterial Revascularization Therapies Study (ARTS). PMID- 11428491 TI - One-year survival following early revascularization for cardiogenic shock (SCHOCK trial). PMID- 11428492 TI - Can we eradicate rheumatic fever in the 21st century? PMID- 11428493 TI - HLA-DQA1 genotyping by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and restriction endonuclease digestion in Papua New Guinea. AB - We have used PCR-SSCP, a technique based on the conformation of single-stranded DNA, to characterize the HLA-DQA1 gene in four geographically diverse population groups in Papua New Guinea. Among the 294 individuals that were studied from Goroka, north coast of Madang, Kimbe and Wanigela, we detected 5 of the 20 known variants of this gene locus. These included alleles 0101, 0102, 0103, 0301 and 0501. Furthermore, variable mobility shifts observed for alleles 0301 and 0501 from Madang suggested a further 3 variants. All 15 combinations of the 5 confirmed alleles were detected and their respective gene frequencies found to be consistent with the groups' ethnic and linguistic diversity. In respect to their frequencies and the observed overall allelic heterozygosity, the distribution in Kimbe showed some similarity to that in the north coast of Madang while Madang and Goroka were the most different. The distribution of alleles 0102 and 0501 was observed to be similar for Goroka and Wanigela as was 0301 for Madang and Wanigela. Our results, confirmed by endonuclease digestion, show PCR-SSCP to be a highly sensitive technique that can be used to characterize HLA-DQ antigens. In addition, the simplicity of the method provides an opportunity for large-scale typing of HLA antigens. PMID- 11428494 TI - The management of splenic trauma in patients with splenomegaly due to malaria. PMID- 11428495 TI - Snakebite and antivenene. PMID- 11428496 TI - Introduction of a new curriculum method of teaching known as problem-based learning to the University of Papua New Guinea Medical School. PMID- 11428497 TI - A survey of pregnant women with tuberculosis at the Port Moresby General Hospital. AB - From March 1995 to February 1998, 110 patients diagnosed with tuberculosis (TB) in pregnancy or the puerperium at the Port Moresby General Hospital (PMGH) were surveyed. 96% were diagnosed as a result of the symptoms of tuberculosis, 4% through contact tracing. 11 of 40 patients who first attended antenatal clinic in the second trimester were not diagnosed until after delivery. The mean birthweight of term infants of TB patients was significantly less than term infants in a previous survey at PMGH. 45% of babies were growth restricted. With increasing duration of treatment, both increasing maternal weight gain in pregnancy and higher mean birthweight were found. Maternal and perinatal mortality were high in the study patients. There were 6 maternal deaths and a perinatal mortality rate of 137/1000. The majority of maternal and fetal losses occurred in patients who had pulmonary, miliary and meningeal TB. Improvement in the detection of tuberculosis in antenatal patients and the introduction of adequate treatment before delivery should prevent maternal deaths and perinatal morbidity and mortality. PMID- 11428498 TI - Reporting of lactation and normal menstrual information by Papua New Guinean women. AB - As part of a survey of 600 women enrolled in a study of postpartum progesterone only contraception (400 women who opted to use progesterone-only contraception and 200 controls) participants were asked about menstrual history and lactation experience. The mean longest menstrual cycle duration was found to be 29.5+/ SD3.5 days and the mean shortest cycle duration was 26.6+/-SD2.8 days. The mean duration of menstrual bleeding was 3.75+/-SD1.16 days. Few women reported menstrual period problems such as dysmenorrhoea (6.5%) and menorrhagia with clots (0.7%). However, 3% of the women reported irregular cycles with intervals of longer than 1 month. Overall the women reported breastfeeding their previous baby for a mean duration of 14 months. The group of women electing to use hormonal contraception reported that they had breastfed their last baby for 13.5+/-SD7.5 months while control women had done so for 14.1+/-SD9.4 months. The longest mean duration that women reported to have breastfed a previous infant was 19.5+/-SD9.6 months in the hormonal contraception group and 19.1+/-SD8.6 months in the control group. PMID- 11428499 TI - Malaria in pregnancy: getting to grips with a sticky problem. AB - In malaria-endemic areas, by young adulthood people have developed functional immunity to malaria. However, during pregnancy this immunity is breached and infection occurs, leading to increased risks for mother and child. In the past this effect has been attributed to nonspecific immunosuppression, but recent research has revealed a specific pathogenic mechanism, involving the adherence of rare parasite variants to the placenta. This commentary explores the current state of research in this area and proposes a model of immune evasion and placental colonization. This model points the way to the development of future therapeutic interventions. PMID- 11428500 TI - Traumatic false aneurysms in Port Moresby. AB - BACKGROUND: Trauma is responsible for about 30% of surgical admissions in Port Moresby. Vascular injuries are frequently missed due to inadequate assessment by primary health care workers and often present late with large aneurysms which are difficult to manage. The aim of this study was to identify the patterns of morbidity associated with traumatic false aneurysms complicating vascular injuries in Port Moresby General Hospital from January 1995 to July 1999. METHODS: A surgical database with 11,004 records was used to identify patients with a diagnosis of false aneurysm. Charts of patients with gunshot wounds, knife wounds and compound fractures were also reviewed to identify further cases with vascular injury. RESULTS: 51 patients were found to have had documented evidence of vascular injury requiring surgical repair within the study period. 21 patients with traumatic false aneurysm presented to Port Moresby General Hospital. 17 of the 21 patients were aged 11 to 40 years. There was only one female patient. Stab wounds (50%) were the commonest mode of injury followed by lacerations (23%) and fish bites (14%). The calf (42%), forearm (13%) and the head (13%) were the commonest sites involved. The time from insult to admission varied from 4 days to 11 years with the modal time being 14 days. Most patients presented with hard signs of false aneurysms (94%) and 81% of the patients were treated within five days. Ligation (67%) was the commonest operation, followed by excision (43%) and vascular repair (19%). There were no deaths or amputations in any of the patients with false aneurysms. CONCLUSION: False aneurysms are a not infrequent complication of vascular injuries. Greater awareness of the possibility of vascular injury is needed. Patients in Port Moresby often present after considerable delay. However, the diagnosis can be made clinically and the results of surgical exploration are good. Although colour Doppler ultrasound is a useful investigation it sometimes fails to identify the aneurysm because it requires blood flow to be present in the aneurysm. PMID- 11428501 TI - Review of cases of hyponatraemia in the Port Moresby General Hospital between August 1993 and June 1995. AB - Hyponatraemia (serum sodium level below 130 mmol/l) is a common electrolyte abnormality in a hospital population. It can be associated with dehydration, overhydration or normal hydration. Clinically, it is important to recognize the common diseases associated with hyponatraemia since correct treatment in terms of fluid replacement is essential in preventing complications of low serum sodium. We have reviewed results of serum sodium tested from patients admitted to the Port Moresby General Hospital between 1993 and 1995. This was aimed at identifying the most common features associated with low sodium. Clinical information and diagnosis were obtained by looking through a series of request forms. Of the approximately 30,000 blood samples taken over 23 months, the percentage of samples with hyponatraemia was about 1%. Hyponatraemia was more common in medical (38%) and paediatric (35%) cases and at the extremes of ages, ie, under the age of 6 years and above 40 years. Over a quarter of the hyponatraemic patients had severe hyponatraemia (serum sodium below 120 mmol/l). Clinical conditions commonly associated with hyponatraemia, in descending order of importance, were diarrhoea and vomiting, renal failure, central nervous system infections and trauma, pulmonary infections, oedematous states (eg, nephrotic syndrome) and diabetes mellitus. PMID- 11428502 TI - Arterial oxygen saturation in healthy young infants in the highlands of Papua New Guinea. AB - To determine the effect of moderate altitude on arterial oxygen saturation (SaO2), pulse oximetry was performed on 302 children aged <3 months attending a clinic in Goroka, Eastern Highlands Province (1584 metres above sea level) for minor ailments or immunization. Respiratory and heart rates were also measured. The overall mean SaO2 was 96%. Comparison between log-transformed means showed that SaO2 was significantly lower in the first month of life than later (p=0.04). 6% of SaO2 values were <92%, which is a practical cut-off for normal SaO2 in this population of highland children aged <3 months. Mean respiratory and heart rates were 50/minute and 145/minute, respectively. After adjusting for age, respiratory rate increased significantly as SaO2 declined (p=0.002). We have thus defined reference values for SaO2, respiratory rate and heart rate in healthy young infants residing in the highlands of Papua New Guinea. Further investigation is needed to determine whether SaO2 is lower in babies when they are asleep and to define reference values for older children in the highlands. PMID- 11428503 TI - Patterns of child growth in Papua New Guinea and their relation to environmental, dietary and socioeconomic factors--further analyses of the 1982-1983 Papua New Guinea National Nutrition Survey. AB - Child growth and nutrition in rural Papua New Guinea vary widely among different environments. The 1982-1983 National Nutrition Survey (NNS) was re-analyzed in order to relate patterns of growth to a wide range of dietary, socioeconomic, agricultural and demographic variables. Anthropometric indices of growth were calculated based on an internal Papua New Guinean growth standard constructed from the children included in the NNS. Children were subsequently classified as stunted, wasted or underweight if they were more than 1 SD below the national mean. Regression analyses on 15,975 children show that variation in growth among environments can largely be accounted for by differences in diet, although significant differences in relation to altitude, relief and rainfall patterns persist. Other important predictors of child growth and nutrition included socioeconomic status, maternal education, marital status of the mother and father's occupation. Variance components analysis revealed that most of the geographical variation in child growth was accounted for by the environmental, dietary, socioeconomic, agricultural and demographic variables included in the regression analyses. Most of the factors which were found to be associated with child growth in this study are related in one form or another to differences in local subsistence agriculture, which may therefore be the main determinant of child growth and nutrition patterns in Papua New Guinea. PMID- 11428504 TI - Removal of anterior clinoid process for basilar tip aneurysm: clinical and cadaveric analysis. AB - The difficulty in the operation for basilar tip aneurysm is the restriction in surgical working space. To resolve this problem, aggressive skull base surgery has been reported, but these techniques are not prevalent. Pterional and subtemporal approaches are commonly used for basilar tip aneurysms. In an attempt to increase the surgical working space during the pterional approach, the anterior clinoid process and the roof of the optic nerve were removed extradurally to increase the mobilization of the intracranial internal carotid artery and optic nerve. The effects of removing the anterior clinoid process and microanatomy in the perioptic area were analyzed by cadaveric procedures in 10 cases (20 sides). With this procedure, the internal carotid artery can be retracted medially with a spatula 6.1 +/- 0.8 mm (mean +/- SD). The length and the area of dural fold in the bone defect region in the optic canal roof are 2.1 mm and 13.6 mm. In 10 clinical cases, this procedure allowed enough space to approach the basilar tip aneurysm without disturbing the internal carotid artery blood flow. The clinical outcome was satisfactory. PMID- 11428505 TI - Cerebral microaneurysms found incidentally during aneurysm surgery. AB - Not uncommonly, cerebral microaneurysms are found incidentally during surgery for another previously diagnosed cerebral aneurysm(s). The frequency and angiographic characteristics of such incidental microaneurysms are retrospectively summarized. Seventeen patients were identified as harboring incidental microaneurysms, comprising 4.9% of the whole series. The middle cerebral artery (MCA) was the most frequent location (seven cases, 41%) of these microaneurysms. There was a tendency for MCA microaneurysms to be contiguous to a previously known, larger aneurysm at the same location. Neurosurgeons as well as interventional neuroradiologists should be aware of the possible presence of these incidental microaneurysms while treating patients with a cerebral aneurysm(s). Although the actual clinical implications of these incidental microaneurysms have not been elucidated, the few additional risks to patients already surgically exposed for the treatment of another aneurysm, along with the possible benefit of preventing their rupture and growth, would justify the surgical treatment of these microaneurysms. PMID- 11428506 TI - Stroke patterns in cardio-embolic infarction in a population-based study. AB - The study demonstrates that clinical-radiological causes and outcome of cardio embolic infarcts in a population-based study correspond to a well-identified stroke pattern. Cardio-embolic infarcts was diagnosed in 882 cases (37.9%) of 2,330 consecutive first-ever stroke patients included in a prospective population based stroke registry over a 14-year period (1985-1997). Thirty-three criteria out of 98 were introduced into a monovariate analysis and the significant variable were introduced into a multivariate analysis to identify significant criteria to define stroke patterns in cardio-embolic infarction. Cardiac sources of embolus included atrial arrhythmia, valvular heart disease (19%), and cardiac failure (18%). Patients with cardio-embolic infarction showed a significantly higher rate of female predominance (p < 0.001), history of ischemic heart disease (p < 0.001), acute stroke onset (p < 0.05), headache (p < 0.05), previous treatment by anti-platelets and anti-K-vitamin (p < 0.001), Wernicke aphasia (p < 0.001), severe deficit (p < 0.001) and death (p < 0.001). After a logistic procedure, female gender and ischemic heart disease were the two independent risk factors associated with cardio-embolic stroke. Cardio-embolic stroke is a specific subtype of stroke with its own clinical, radiological, etiological and prognostic characteristics. In the acute stage, it is necessary to identify quickly this type of stroke because of severe prognosis and appropriate treatment. PMID- 11428507 TI - Advanced neuromonitoring including cerebral tissue oxygenation and outcome after traumatic brain injury. AB - For 51 patients suffering from traumatic brain injury (GCS < 9), we compared the prognostic value of critical parameters derived from neuromonitoring of intracranial pressure (ICP), cerebral perfusion pressure (CPP) and brain tissue oxygenation (PiO2) during different time periods after trauma (< or = 12, < or = 24, < or = 48, < or = 72 and < or = 96 h). For patients with good outcome (GOS = 4-5, n = 30) the proportion of critical ICP values (> 40 mmHg) was about 0.2% during all time periods. The corresponding proportions for patients with bad outcome (GOS = 1-3, n = 21) rose from 0.2% to 4.7% during increasing time periods. The frequency of critical ICP values was significantly related to outcome (p < 0.001) for time periods > 48 h after trauma. Differences of critical CPP (< or = 50 mmHg) and hypoxic PiO2 (< or = 5 mmHg) between both outcome groups were less pronounced and for both parameters significant relations to outcome were only obtained for the longest time period (< or = 96 h, p < or = 0.05). Higher thresholds for CPP (< or = 60 mmHg, < or = 70 mmHg) did not reveal any relation to outcome. For all neuromonitoring parameters significant relations between the frequency of critical values and outcome could be determined. Critical ICP values provide the earliest and highest prognostic power, while critical CPP and hypoxic PiO2 only showed prognostic power in later time periods. PMID- 11428508 TI - An efficient method for the culturing and generation of neurons and astrocytes from second trimester human central nervous system tissue. AB - The isolation, culturing and expansion of human neural progenitors cells has important potential clinical applications in cellular transplantation strategies as well as in developmental studies involving the central nervous system (CNS). This study describes an efficient method to culture neurons and astrocytes as primary cultures, as well as from proliferative progenitor cells derived from second trimester fetal CNS tissue. Second trimester fetal human tissue was mechanically dissociated and subjected to trypsin-dissociation and trituration. The resulting suspension was passed over a Percoll density gradient. The middle (second) fraction of cells was centrifuged to yield a homogenous population of cells with 80%-90% viability. These cells were either cultured directly on laminin coated dishes with defined medium supplemented with fetal bovine serum or in defined medium supplemented with growth factors including epidermal growth factor, basic fibroblast growth factor and leukemia inhibitory factor. The primary cell cultures yielded neurons and astrocytes after 3-5 days in vitro verified by immunostaining with MAP2ab and GFAP. Cells exposed to growth factor supplemented medium formed free-floating spheres within one week. Upon growth factor removal and plating on laminin-coated dishes, brain derived spheres gave rise to neurons, astrocytes and oligodendrocytes; spinal cord derived spheres generated only astrocytes. This protocol describes an efficient method to generate and culture neurons and astrocytes from second trimester human CNS tissue that may be useful in transplantation and developmental studies. PMID- 11428509 TI - Time courses of increased expression of signaling transduction molecules induced by basic fibroblast growth factor in PC12 cells. AB - We previously demonstrated that basic fibroblast growth factor (bFGF) protected neuronal injury in in vivo experimental cerebral ischemia. The precise molecular mechanisms of the neuroprotective effect of bFGF, however, remains unsolved. We investigated time courses of up-regulated molecules involved in intracellular signaling transduction pathways induced by bFGF in PC12 cells to explore the possible neuroprotective mechanism of bFGF action. In Western blot analysis, bFGF increased expression of Ras mainly in the early stage up to 24h, returning to the baseline level at 48 h. Expression of phosphatidylinositol 3-kinase (PI 3-kinase) was enhanced throughout the early and later stages, and was more up-regulated at 48 h compared to 24 h. The present findings suggest that bFGF might promote cell survival or proliferation mainly via Ras, and PI 3-kinase might be involved in cell survival and differentiation in PC12 cells. PMID- 11428510 TI - Microsurgical anatomy of the ophthalmic artery and the distal dural ring for the juxta-dural ring aneurysms via the pterional approach. AB - Microsurgical anatomy for the pterional approach was studied regarding the origin and the course of the ophthalmic artery and the distal dural ring using human cadaveric specimens, with special reference to the surrounding bony structures. In 50 human adult formalin-fixed cadaveric cerebral hemispheres and 10 block specimens of the skull base region including the ophthalmic artery and the carotid dural ring were examined under magnification using an operating microscope. The ophthalmic artery originated from the intradural portion of the internal carotid artery (ICA), except in 5% where the ophthalmic artery originated extradurally. The extradural origin had two patterns: one was that the ophthalmic artery penetrated the bony optic strut (trans-optic strut pattern) and the other was that it coursed into the optic canal proximally to the optic strut without bone penetration (supra-optic strut pattern). The origin of the intradural ophthalmic artery was commonly located at the medial third of the superior wall of the ICA (78%). The ophthalmic artery was commonly taking an S shaped course in the intradural portion and entered the optic canal over the optic strut. The distal dural ring was tightly adherent to the internal carotid artery; circumferential sectioning of the dural ring is required to mobilize the internal carotid artery. When approaching juxtadural ring ICA aneurysms via the pterional route, it is important to recognize the extradural origin, especially the trans-optic strut type, and to precisely understand the microsurgical anatomy around the dural ring. PMID- 11428511 TI - Secondary partial empty sella syndrome in an elite bodybuilder. AB - The pituitary gland is a hormone-responsive gland and is known to vary in size depending on the hormonal status of the patient and the multifaceted positive and negative feedback hypothalamic-pituitary-gonadal axis. Partial empty sella syndrome with an atrophied pituitary gland is seen in primary neuroendocrinopathies such as growth hormone deficiency, primary hypothyroidism, central diabetes insipidus and hypogonadism. Partial empty sella has also been shown to occur in patients with elevations in intracranial pressure. Secondary partial empty sella syndrome with significant pituitary gland atrophy from negative feedback inhibition of long-term exogenous hormonal use has not been previously reported. We are reporting on a case of partial empty sella syndrome occurring in an elite bodybuilder with a long history of exogenous abuse of growth hormone, testosterone and thyroid hormone. The pathophysiological mechanisms of secondary partial empty sella syndrome from exogenous hormone use and the possibility for elevations in intracranial pressure contributing to this syndrome will be discussed. PMID- 11428512 TI - Method to improve cosmetic outcome following craniotomy. AB - This technical note describes a simple method for reducing the dead space created by craniotome due to the loss of bone dust and improving the cosmetic outcome following a craniotomy. After drilling the burr holes for the craniotomy, the bone between the holes is drilled away in a standard fashion except that multiple regions of about 1 cm in length are left intact. These intact regions are broken using a periosteal elevator and fixed like a bridge when the bone is replaced. The resulting bone flap is readily returned to its original position without making the dead space created by regular craniotomy. The amount of the dead space caused by losing the bone dust is reduced and a good cosmetic recovery is obtained. This technique is useful for both craniotomy and facial bone surgery, which requires cosmetic results. PMID- 11428513 TI - Transcranial magnetic stimulation and acoustic trauma or hearing loss in children. AB - Transcranial magnetic stimulation is a non-invasive method used to assess motor function in humans; however, some reports suggest it may cause internal ear damage (cochlear). Eighteen patients with normal auditory function (ages 2 months to 16 years, mean 6.8 years), two medical doctors and two technicians who performed the studies were tested with brain stem auditory evoked potentials, otoacoustic emissions, acoustic reflex and a pure tone audiometric and logoaudiometric test when possible, before and after transcranial magnetic stimulation for central motor conduction studies in different neurological conditions. All the tests were repeated two weeks and two months later. Patients had no auditory protection nor history of seizures. Motor evoked potentials and silent periods were recorded from the right abductor pollicis brevis and the first dorsal interosseous muscles at rest and during weak voluntary contraction when possible. A mean of 48 transcranial magnetic stimulations with 50%-75% Tesla intensity were used. Natural logarithmic transformation of latency and amplitude data had a normal distribution. There were no significant differences in auditory function testing. PMID- 11428514 TI - Spinal cord sarcoidosis: a diagnostic dilemma. AB - Sarcoidosis is a chronic disorder of unknown etiology characterized by the development of non-caseating granulomas with derangement of the normal tissue architecture. Compromise of the spinal cord is one of the rarest neurologic manifestations of the disease, which may be clinically and radiologically indistinguishable from a spinal cord malignant tumor. However, neurosarcoidosis can be treated with steroids. This study reviews the clinical, radiological, and pathological features of the sarcoid compromise of the spinal cord, emphasizing the difficulties commonly encountered in making a diagnosis. PMID- 11428515 TI - FGF-2 immunoreactivity in adult rat ependyma and choroid plexus: responses to global forebrain ischemia and intraventricular FGF-2. AB - Fibroblast growth factor 2 (FGF-2) immunoreactivity (IR) was examined in the ependyma and choroid plexus (CP) of lateral and third ventricles in normal adult rats, as well as in response to transient forebrain ischemia (TFI) and exogenous FGF-2 delivered intraventricularly for several days by osmotic pump. Similar patterns of FGF-2 IR were seen in the CP epithelia of both lateral and third ventricles, as well as in ependymal cells of the third ventricle and along lateral sides of the lateral ventricles. Consistent staining was seen along the apical aspect of epithelial cells facing the cerebrospinal fluid (CSF). Cytoplasmic staining was seen in the absence of ischemia, and was dramatically reduced in response to TFI. FGF-2 treatment followed by TFI resulted in sustained FGF-2 IR within CP and ependymal cells, supporting the idea that these tissues are involved in synthesis and secretion of growth factors into the CSF. In contrast, along the medial sides of the lateral ventricles, adjacent to brain structures such as the hippocampus, consistent staining was seen along the basal aspect of the ependymal cells. We propose that at least some regions of ependyma may function to transport molecules such as FGF-2 directly into the underlying brain parenchyma. PMID- 11428516 TI - Modified immunoregulation associated with interferon-gamma treatment of rat glioma. AB - Little is known about modulation by cytokines of major histocompatibility complex (MHC) antigen expression on intracranial tumors in vivo. The ability of cytokines to up-regulate MHC class-1 (MHC-1) antigen expression was investigated first in vitro using three rat glioma cell lines. Immunohistochemistry showed that incubation with recombinant rat interferon-gamma (rrIFN-gamma) increased MHC-1 antigen expression in RG2, C6, and 9L cell lines. Flow cytometric analysis revealed different baseline levels of MHC-1 antigen expression in each line (RG2 lowest, C6 highest), and that these levels increased in all lines after stimulation with 100 U ml(-1) or more of rrIFN-gamma. The antitumor effect of rrIFN-gamma in vivo was evaluated by assessing survival of rats with implanted intracerebral RG2 gliomas after intracarotid infusion of rrIFN-gamma. A high dose of rrIFN-gamma (2.4 x 10(5) U kg(-1)) significantly increased the survival, compared to control (p < 0.02). Intracarotid pre-treatment with the bradykinin analogue RMP-7 did not further increase survival. Immunohistochemical staining of tumor sections after in vivo rrIFN-gamma, infusion showed no clear increase in MHC-1 antigen expression on tumor cells but increased staining for ED2 antigen within tumor tissue, presumably from perivascular cells with MHC class-2 antigen. PMID- 11428517 TI - Mechanism of hemolysate-induced [Ca2+]i elevation in cultured fibroblasts. AB - Erythrocyte lysate (hemolysate) released from blood clot after subarachnoid hemorrhage is the causative agent for chronic cerebral vasospasm, a prolonged contraction of cerebral arteries. Fibroblasts, the outer layer cells of vessel wall that in contact with blood clot directly, may contribute to cerebral vasospasm. However, the effect of hemolysate on intracellular Ca2+ ([Ca2+]i) mobilization in fibroblasts has not been studied. We investigated hemolysate induced [Ca2+]i mobilization in cultured neonatal human dermal and canine middle cerebral arterial fibroblasts by using fura-2 microfluorimetry. Hemolysate increased [Ca2+]i by releasing internal Ca2+ stores and promoting Ca2+ entry. Tyrosine kinase inhibitors partially but significantly reduced the effect of hemolysate. The major components of hemolysate, oxyhemoglobin (OxyHb) and adenosine triphosphate (ATP) failed to mimic the effect of hemolysate. In cultured canine middle cerebral arterial fibroblasts, hemolysate produced similar Ca2+ mobilization to that of dermal cells. OxyHb and ATP failed again to reproduce the effect of hemolysate. We conclude that hemolysate increases [Ca2+]i in fibroblasts and this effect of hemolysate is not mediated by OxyHb or ATP but by some unknown factors. PMID- 11428518 TI - Internal shunting in small vessel reconstruction: an experimental study. AB - We describe the experimental use of an endoluminal shunt in different procedures for small vessel reconstruction. Since the arteries used in this work are similar in diameter to those of the brain, this method could be applied to human vascular surgery. PMID- 11428519 TI - Correlation between bradykinin-induced blood-tumor barrier permeability and B2 receptor expression in experimental brain tumors. AB - Localization of B2 receptors in brain tumor cells and microvessel endothelial cells of the brain tumors was investigated to study the differential sensitivity of brain tumors to bradykinin. The present study shows that B2 receptor expression levels vary in cultured RG2, C6 and 9L glioma cells as well as in the intracerebral tumors established with these cell lines in rats. The double immunohistochemical data indicate that B2 receptors are localized to tumor cells and not to the tumor capillaries. Immunostaining and Western blot analysis for B2 receptor showed that the B2 receptor expression was in the order C6 > RG2 > 9L. The permeability studies on RG2, C6 and 9L tumors in rats showed that C6 tumor had the highest increase (178%) in Ki (unidirectional transport across blood brain barrier (BBB)/blood-tumor barrier (BTB)), while 9L tumor had the least increase of Ki (35%) over the control group, following intracarotid infusion of bradykinin. We found a positive correlation (r = 0.965, p < 0.001) between B2 receptor levels and bradykinin-induced increase in BTB permeability. We conclude that B2 receptors are localized to tumor cells and not to normal or tumor capillary endothelial cells. C6 tumor with highest B2 receptor expression was most responsive to bradykinin, while RG2 and 9L tumors with lower B2 receptor expression level were less sensitive to bradykinin with regard to BTB permeability. PMID- 11428520 TI - Strategy for the treatment of inaccessible unruptured giant and large aneurysms of the internal carotid artery. AB - Our goal was to clarify the optimum management of the inaccessible unruptured giant and large aneurysms of the internal carotid artery (ICA). Since 1981, we have treated 18 patients with unclippable unruptured giant or large aneurysms of the ICA. Aneurysms were classified as either intracavernous or intradural. We performed proximal carotid occlusion in 12 patients and conservatively treated six patients. We retrospectively analyzed long-term outcomes in these patients. Four of seven patients with intradural aneurysm underwent proximal carotid occlusion, with good long-term outcomes. The three patients with intradural aneurysm, who were treated conservatively, died of subarachnoid hemorrhage. Eight of 11 patients with intracavernous aneurysm underwent proximal carotid occlusion, one dying of massive nasal bleeding 25 months after the procedure. In this case, the aneurysm was partially thrombosed, and residual lumen growth was revealed 22 months after proximal carotid occlusion. Cranial nerve paresis improved in five of the eight patients (63%), and two patients had a minor ischemic attack. Neurological problems failed to occur in the three patients with intracavernous aneurysm who were treated conservatively. The risk of rupture is relatively high in intradural giant and large aneurysms. Proximal carotid occlusion can effectively prevent bleeding from intradural aneurysms. Aggressive management is justified for intradural aneurysms with poor collateral circulation. Operative procedures in the management of an intracavernous aneurysm require careful consideration. PMID- 11428521 TI - Fractal dimension analysis of static stabilometry in Parkinson's disease and spinocerebellar ataxia. AB - The static stabilometry patterns associated with Parkinson's disease (PD, n = 15) and spinocerebellar ataxia (SCA, n = 15) were compared with those of normal control (n = 15) by measuring the fractal dimensions. Fractal dimensions were estimated using the modified pixel dilation (mPD) method. The fractal dimensions with closed eyes showed a significant correlation with Environmental area for SCA group (p < 0.05). The fractal dimension for SCA group was significantly higher with closed eyes than that with open eyes (p < 0.05). The fractal dimension with closed eyes was significantly higher in PD and SCA groups than that in normal group (p < 0.05). The fractal dimension with closed eyes was higher when the clinical stage was more severe with PD and SCA group while Environmental and Longitude/Environmental areas were not. These findings suggest that the fractal dimension is more sensitive than traditional stabilometric analysis in an evaluation of postural instability in PD and SCA. PMID- 11428522 TI - Acrylic hydrogel implants after spinal cord lesion in the adult rat. AB - Acrylic hydrogels, like the polymer of 2-hydroxyethyl methacrylate, are biocompatible, mechanically stable, porous materials that can be coated with collagen or laminin acting as bioadhesive substrates. Poly-2-hydroxyethyl methacrylate sponges have been proposed for restoring the anatomical continuity of damaged neural structures. In the present work, the ability of poly-2 hydroxyethyl methacrylate sponges to provide the injured spinal cord neurons with a conductive substrate for their regenerating axons was investigated in 32 adult Wistar rats. Collagen impregnated poly-2-hydroxyethyl methacrylate sponges were implanted into suction cavities of the dorsal funiculus of the spinal cord. Two to four months after implantation, the spinal cord was removed and processed for histology, and S100 and GFAP immunohistochemistry. To study axonal regeneration into the sponge, the spinal cord or the sensorimotor cortex were injected with 0.05-0.1 microl of an 8% solution of lectin-conjugated horseradish peroxidase or 10% dextran tetramethylrhodamine. The fibroglial reaction, accumulation of mononuclear cells, and angiogenesis at the interface between the spinal cord and the sponge were minimal. Cystic cavitation in the spinal cord was virtually absent. Anterograde labeled axons were seen to penetrate and to elongate the full length of the sponge. These results demonstrate that poly-2-hydroxyethyl methacrylate sponges represent a safe supportive material for regenerating spinal cord axons. PMID- 11428523 TI - 3-Aminobenzamide, a poly (ADP-ribose) synthetase inhibitor, attenuates the acute inflammatory responses and brain injury in experimental Escherichia coli meningitis in the newborn piglet. AB - The aim of the present study was to evaluate the anti-inflammatory and neuroprotective effects of a poly (ADP-ribose) synthetase inhibitor 3 aminobenzamide during the early phase of experimental bacterial meningitis in the newborn piglet. Meningitis was induced by intracisternal injection of 10(8) colony forming units of Escherichia coli in 100 microl of saline. 3 Aminobenzamide, given 30 mg kg(-1) as a bolus i.v. injection 30 min before induction of meningitis, significantly attenuated the meningitis-induced acute inflammatory responses such as increased cerebrospinal fluid (CSF) lactate concentration, CSF leukocytosis and increased CSF tumor necrosis factor-alpha level. However, meningitis-induced increase in intracranial pressure and decrease in CSF glucose level were not significantly improved. Increased cerebral cortical cell membrane lipid peroxidation products (conjugated dienes) and decreased brain ATP/phosphocreatine levels observed in the meningitis group were also significantly improved with 3-aminobenzamide treatment. However, the improvement of reduced Na+, K+-ATPase activity did not reach a statistical significance (p = 0.06). In summary, 3-aminobenzamide significantly attenuated the acute inflammatory responses and the ensuing brain injury during the early phase of neonatal bacterial meningitis. These findings suggest that poly (ADP-ribose) synthetase inhibitors such as 3-aminobenzamide might be a promising novel anti inflammatory and neuroprotective adjuvant therapy in neonatal bacterial meningitis. PMID- 11428524 TI - Hepatocyte growth factor reduces the infarct volume after transient focal cerebral ischemia in rats. AB - Hepatocyte growth factor (HGF) was originally discovered as a powerful mitogen for hepatocytes. HGF also has been reported to function as a neurotrophic factor as well as an angiogenetic factor. The present study examined the neuroprotective effect of HGF against transient focal cerebral ischemia in rats, in which an anti apoptotic and an angiogenetic effect of HGF was assumed to contribute to the reduction of the infarct volume. The intraventricular administration of human recombinant HGF prevented neuronal death after 120 min of occlusion in the right middle cerebral artery and the bilateral common carotid arteries. HGF significantly reduced the infarct volume in a dose-dependent manner. In a separate series of experiments, we next histopathologically investigated both the anti-apoptotic effect on neurons and the angiogenetic effect of HGF. A large number of TUNEL positive neurons were observed in the inner boundary of the infarct area in both the control and the vehicle group whereas only a few TUNEL positive neurons were observed in the corresponding area in the HGF group. In the HGF group, Bcl-2 protein was obviously represented in surviving neurons subjected to ischemia. The number of the vascular lamina in HGF group were significantly higher than those in the vehicle group. These data suggest that HGF appears to have an ability to prevent apoptotic neuronal cell death while also possessing an angiogenetic effect in the central nervous system which was affected with transient focal cerebral ischemia. PMID- 11428525 TI - What are you reading? PMID- 11428526 TI - An inventory of information on blindness and visual impairment in Canada. AB - BACKGROUND: Several health databases contain information on blindness and visual impairment in Canada. Such databases may permit studies of the outcomes, predictors, economic costs and meta-analysis of ocular health problems and visual disability. In this paper we summarize the existing public health information on blindness and visual impairment in Canada. METHODS: A systematic search was performed to find the information in three areas: health administrative databases, health surveys and registries. Both literature searching and Internet searching were performed. The institutions were notified by email or telephone that information pertaining to vision and blindness was desired. When necessary, we visited the institution to obtain the information. RESULTS: Health administrative databases contain information on a wide cross-section of diagnoses but are usually limited in detail and seldom provide longitudinal information. Health surveys have a limited amount of systematic information on vision-related questions and rely exclusively on self-reporting. Registries exist on a wide range of visual topics. The Canadian National Institute for the Blind (CNIB) has the most comprehensive registry. INTERPRETATION: This is the first attempt at summarizing all the public health information on blindness and visual impairment in Canada. The present population-based ocular information has numerous shortcomings. More comprehensive validation of large registries, such as that of the CNIB, is needed to provide the foundation for a longitudinal ocular surveillance system. Such a system could guide research on risk factors and the effect of treatment, economic analyses, and public policy for ocular research and service allocation. PMID- 11428527 TI - Identification of diabetic retinopathy by stereoscopic digital imaging via teleophthalmology: a comparison to slide film. AB - BACKGROUND: Diabetic retinopathy is a leading cause of vision loss in North America. We compared mydriatic seven-field stereoscopic digital imaging to 100 ASA slide film photography for the identification of diabetic retinopathy via teleophthalmology. METHODS: Patients from a northern Alberta community with diabetes mellitus diagnosed by a physician were asked to participate in a teleophthalmology pilot project. Patients were enrolled at four different times between October 1999 and June 2000. Seven 30 degree fields of the retina were photographed with both slide film and digital imaging (resolution 2008 x 3040 pixels) through a dilated pupil. Slide film was developed and reviewed in a masked fashion by a retinal specialist. Digital images for each patient were transmitted by satellite to Edmonton and analysed a minimum of 2 months after the original slide film. Retinal abnormalities were graded with the use of the Early Treatment Diabetic Retinopathy Study extension of the modified Airlie House classification. We calculated the sensitivity and specificity of digital imaging for the identification of features of diabetic retinopathy as seen on slide film. Pearson's correlation coefficient was also calculated. RESULTS: A total of 121 patients (241 eyes), of whom 114 (94.2%) had non-insulin-dependent diabetes, participated in the study. The average duration of diabetes was 8.5 years. Of the 121 patients 57 (47.1%) had diabetic retinopathy, 12 (9.9%) had clinically significant macular edema and 2 (1.6%) had neovascularization. Pearson's correlation coefficient for the presence of retinopathy between slide film and stereoscopic digital imaging was 0.92 for microaneurysms, 0.80 for hemorrhages, 0.45 for intraretinal microvascular abnormalities, 0.32 for venous beading, 1.00 for neovascularization of the disc, 1.00 for neovascularization elsewhere in the retina and 0.97 for clinically significant macular edema (p < 0.001). The correlation between the two techniques for severe nonproliferative diabetic retinopathy (NPDR) was 0.86 and for high-risk proliferative diabetic retinopathy 1.00 (p < 0.001). INTERPRETATION: Stereoscopic digital imaging has a high level of correlation with slide film for the identification of most features of diabetic retinopathy, including microaneurysms, hemorrhage, severe NPDR, high risk proliferative diabetic retinopathy and clinically significant macular edema. PMID- 11428528 TI - Contrast sensitivity and depth of focus with aspheric multifocal versus conventional monofocal intraocular lens. AB - BACKGROUND: Laboratoires Domilens, Lyon, France, has introduced a new aspheric multifocal intraocular lens (IOL), the Progress 3. The central portion, measuring 4.7 mm in diameter, has an anterior surface of progressively increasing power, such that there is a central add of +5.00 dioptres. We compared contrast sensitivity and depth of focus in patients who received the Progress 3 and in those who received a conventional monofocal IOL. METHODS: Prospective study. Forty patients with age-related cataract were randomly divided into two groups: 20 patients received the Progress 3 aspheric multifocal IOL, and 20 patients received a conventional monofocal IOL of similar design. Contrast sensitivity was measured with the Pelli-Robson letter-based chart. Depth of focus was determined by dialling a series of overcorrections over the patient's manifest refraction until the patient read 6/12 clearly. The depth of focus was defined as the range over which 6/12 or better acuity was achieved. Quality of vision was evaluated by patient questionnaire. RESULTS: Mean contrast sensitivity was significantly lower in the patients with a multifocal IOL than in those with a monofocal IOL (1.38 vs. 1.56 log units) (p < 0.001). The mean depth of focus values for the two groups were 3.10 D and 1.65 D respectively (p < 0.001). The prevalence of subjective problems was similar in the two groups. INTERPRETATION: In our opinion, aspheric multifocal IOLs should be reserved for patients who are willing to trade increased depth of focus for reduced contrast sensitivity postoperatively. PMID- 11428529 TI - Reliability of the time trade-off technique of utility assessment in patients with retinal disease. AB - BACKGROUND: Studies in medical fields other than ophthalmology have given conflicting results regarding the reliability of the time trade-off technique of utility assessment. We performed a study to determine the test-retest reliability of the time trade-off technique for assessing utilities in patients with ocular diseases of the retina and to investigate possible factors associated with differences in utility over time. METHODS: Patients referred to the retina service of a tertiary care hospital in eastern Canada were eligible for the initial interview if they had best corrected vision of 20/30 or worse in at least one eye and were deemed competent to answer the required questions. Patients were interviewed prospectively between December 1999 and March 2000 during a normal 30 minute period needed for pharmacologic mydriasis to occur. Demographic, clinical (including Snellen visual acuity) and time trade-off utility information was collected through chart review and standardized interview. Patients who completed the interview successfully were called back 28 days later for follow-up. RESULTS: Of the 138 eligible patients 112 (81.2%) completed the initial interview. Of the 112, 96 (85.7%) completed the second interview. Half of the respondents were women, and all but one respondent were white. The mean age was 65.3 years. The primary reasons for visual loss included diabetic retinopathy (59 patients [61.4%]) and age-related macular degeneration (14 patients [14.6%]). The intraclass correlation coefficient between the initial and follow-up visual utilities was 0.7634 (95% confidence interval 0.6655-0.8355). INTERPRETATION: Our results show excellent reliability of the time trade-off technique of utility assessment in patients with ocular diseases of the retina. PMID- 11428530 TI - Successful spectacle correction for an ametropic amblyopic child with congenital auricular aplasia using a new spectacle-supporting apparatus. PMID- 11428531 TI - Lacrimal gland inflammation as the presenting sign of Crohn's disease. PMID- 11428532 TI - Resolution of bilateral multifocal subretinal cysticercosis without significant inflammatory sequelae. PMID- 11428533 TI - Total removal of an unruptured orbital hydatid cyst. PMID- 11428534 TI - Lowering of blood pressure and coronary blood flow in isolated systolic hypertension. AB - BACKGROUND: In essential hypertension, the lower limit of autoregulation of coronary flow shifts to higher perfusion and the hypertensive ventricle is at a higher than normal risk of ischemia, and less able to tolerate acute reduction of coronary perfusion pressure. Little is known about pattern of coronary flow in isolated systolic hypertension, a pathologic condition in which the elevated systolic blood pressure is associated with a lower than normal vascular compliance and normal or slightly greater than normal mean arterial pressure and vascular resistance. OBJECTIVE: To evaluate the effects of rapid normalization of blood pressure on coronary blood flow in isolated systolic hypertension. METHODS: We subjected 20 patients with isolated systolic hypertension to intraoperative hemodynamic and transesophageal echocardiographic monitoring during peripheral vascular surgery. Coronary flow velocity integrals and diameters in the left anterior descending coronary artery were evaluated under baseline conditions and after normalization of blood pressure, which occurred spontaneously during anesthesia (10 cases; group 1A) or was induced by infusion of nitrate (10 cases, group 1B). RESULTS: After normalization of systolic blood pressure integrals decreased significantly only for patients in group 1A; percentage changes of diameter were significantly greater for patients in group 1B. Therefore, coronary blood flow after normalization of systolic blood pressure increased for patients in group 1B (by 28+/-25%) and decreased for patients in group 1A (by 30+/-21%). Changes in integrals were inversely related to those in diameter (r= -0.72, P < 0.001); for patients in group 1A changes in coronary perfusion pressure and diameter were related to those of integrals (r= 0.94; P < 0.0005). CONCLUSIONS: In isolated systolic hypertension, despite there being similar changes of the systolic blood pressure, administration of nitrates caused a marked increase of coronary flow through direct effects on coronary circulation, whereas spontaneous normotension was associated with a significant reduction of coronary flow. PMID- 11428535 TI - Cyclic variation of the myocardial integrated backscatter signal in hypertensive cardiopathy: a preliminary study. AB - BACKGROUND: Ultrasound tissue characterization studies realized through integrated backscatter analysis with end-diastolic sampling in hypertensive cardiopathy have demonstrated that abnormalities in the left ventricular myocardial ultrasonic texture are present in extreme forms of left ventricular hypertrophy (LVH). Such abnormalities are not evident in the athlete's heart. The aim of the present study was to analyze the ultrasonic backscatter myocardial indexes both as peak end-diastolic signal intensity and as cardiac-cyclic variation in two models of LVH: hypertensive cardiopathy and athlete's heart. METHODS: Three groups of 10 subjects each, all men of mean age (31.6+/-3.5 years), and of comparable weight and height, were analyzed. Group A comprised 10 cyclists of good professional level, while hypertensive patients were grouped in Group H. Both groups presented a comparable left ventricular mass (LVM). Group C included 10 healthy subjects acting as controls. The men with hypertension were selected on the basis of the results of ambulatory monitoring of the blood pressure according to ISH-World Health Organization guidelines (International Society of Hypertension). A 2D-color Doppler echocardiography with a digital echograph Sonos 5500 (Agilent Technologies, Andover, Massachusetts, USA), was carried out on all the subjects in the study for conventional analysis of the LVM and function. The ultrasonic myocardial integrated backscatter signal (IBS) was analyzed with an 'acoustic densitometry' module implemented on a AT echograph. The signal was also sampled with a region of interest (ROI) placed at interventricular septum and at posterior left ventricular wall level. The systo diastolic variation of the backscatter was also considered, as cyclic variation index (CVIibs). RESULTS: According to the inclusion criteria, the LVM was comparable in groups A and H, but it was significantly higher than group C (left ventricular mass (body surface) (LVMbs)=154.5+/-18.7 (A), 146.8+/-25.5 (H), 101.4+/-12.4 (C), p < 0.001). The end-diastolic IBS did not show significant statistical differences among the three groups. The CVI(IBS) both at septum (30.5+/-5.3 (A), 13.2+/-13.1 (H), 27.2+/-7.3(C), p < 0.002) and posterior wall level (43.7+/-9.1 (A), 16.5+/-12.1 (H), 40.7+/-9.1 (C), p < 0.001) though, was significantly lower in the hypertensive patients than in both the athletes and the control group, where the results were comparable. CONCLUSION: A significant alteration of the myocardial CVIibs (both for septum and posterior wall) was found in the hypertensive model. This was probably the expression of an alteration in the intramural myocardial function. PMID- 11428536 TI - Clinical implications of early ST-segment variability. A report from the ASSENT 2 ST-monitoring sub-study. AB - OBJECTIVES: Evolution of the ST-segment during acute myocardial infarction has been shown to yield more information on prognosis than invasive measurements. By continuous ST-monitoring even very occasional dynamic changes can be analysed. We have recently suggested these variations to be of prognostic importance and possibly reflect individual abilities to deal with a vascular event. We wanted to confirm these findings. METHODS: Four hundred and forty-eight patients were included in the vectorcardiographic sub-study of the second Assessment of Safety and Efficacy of a New Thrombolytic (ASSENT 2) trial. Patients underwent 24 h of ST-monitoring. ST-trend curves were blindly analysed by two independent observers. RESULTS: ST-variability, defined as an increase of the ST-segment shift of > or = 25 microV for 2 min or more, was found to predict death, reinfarction at 30 days or urgent revascularization. By combining variability with resolution of the ST-segment elevation we could identify a high-risk group with 9.9%, and a low-risk group with only 0.8% 30-day mortality. Hypertensive patients, suggested to have an impaired secretion of endogenous t-PA, expressed significantly more ST-variability, possibly a non-invasive marker of impaired capability of dissolving and preventing thrombi. CONCLUSION: Small variations in ST-segment shift during the first 4 h of acute myocardial infarction predict worse outcome. PMID- 11428537 TI - Time course of vascular remodeling, formation of neointima and formation of neoadventitia after angioplasty in a porcine model. AB - BACKGROUND: Vascular remodeling is the major cause of restenosis after coronary balloon angioplasty but the pathophysiology of this process is not known. OBJECTIVE: To examine the time courses of vascular remodeling, formation of neointima and adventitial changes after coronary angioplasty. DESIGN: An experimental study on pigs using coronary angiography, intravascular ultrasound (IVUS), and histology. METHODS: Deep vessel-wall injury was induced by conventional balloon angioplasty in the circumflex and right coronary arteries, and by retraction of a chain-encircled balloon in the left anterior descending artery. Angiography in all three arteries and IVUS measurements in circumflex and left anterior descending arteries were performed before and after angioplasty, and at follow-up on days 0, 1, 4, 7, 14, 28, and 56 (n = 5 in each group). Serial IVUS measurements were used to determine vascular remodeling. Formation of neointima and neoadventitia was measured by histomorphometry. RESULTS: Angiographically evident loss of lumen and ultrasonographically detectable constrictive remodeling occurred between day 7 and day 28. IVUS measurements showed that late loss of lumen (days 28 and 56) was correlated to vascular remodeling but not to the increase in wall area (neointima plus media). Histomorphometry revealed that neointima was present from day 7 and that amount of neointima increased up to day 28. Area of adventitia increased during the first 4 days and remained unchanged thereafter. Adventitial neovascularization by vasa vasorum was observed from day 4 onward. CONCLUSIONS: Formation of neoadventitia precedes late loss of lumen, constrictive remodeling, and formation of neointima. The time course of vascular remodeling coincides with growth of neointima rather than with changes in the adventitia. PMID- 11428538 TI - New non-invasive protocol for detection of coronary spastic angina with significant organic stenosis. AB - OBJECTIVES: This study sought to determine whether a newly-combined test, accelerated exercise following mild hyperventilation (HV) is more beneficial to detect ischaemic evidence in patients with pharmacology-induced coronary artery spasm (CAS) and luminal narrowing of > 75% than classic methods. METHODS AND RESULTS: Forty consecutive patients who all had luminal narrowing of > 75% but < 90% and pharmacology-induced coronary vasospasms of fixed lesions were involved in this study. In these patients, initial HV test, followed by treadmill (TM) exercise test and lastly the newly combined test were performed on three consecutive days. Of the 40 patients, firstly six, secondarily 16 and lastly 32 had positive responses to the HV test, TM exercise test, and newly combined test, respectively. The remaining six patients (15%) had negative results, although the triple sequential tests were performed. Thus, sensitivity of the HV test, the TM exercise test, and the newly combined test was 15% (6/40), 40% (16/40), and 84% (32/38), respectively. Specificity of the three tests were all 100% (46/46). Non sustained ventricular tachycardia and hypotension were observed in two (5%) patients. However, no serious or irreversible complications were encountered in this study. CONCLUSIONS: We recommend the newly combined protocol rather than the classic tests for the detection of ischaemic evidence in patients with coronary spastic angina and fixed stenosis. PMID- 11428539 TI - Non-lipid effects of statin on hypercholesterolemic patients established to have coronary artery disease who remained hypercholesterolemic while eating a step-II diet. AB - BACKGROUND: Results of clinical trials of statin therapy demonstrate that an improvement in incidence of cardiovascular end points and coronary stenosis can be achieved. The beneficial effects of statins on clinical events may involve nonlipid mechanisms that affect endothelial function, such as inflammatory responses, formation of thrombi, and stabilization of plaque. OBJECTIVE: To investigate levels of serologic markers, which may be useful surrogates for activity of vascular disease after administration of statin. METHODS: We administered 20-40 mg simvastatin daily for 14 weeks to 13 patients established to have coronary artery disease who remained hypercholesterolemic during step-II diet therapy. RESULTS: Administration of simvastatin significantly lowered lipoprotein levels and the low: high-density lipoprotein cholesterol level ratio and apolipoprotein B:A-I level ratio compared with pretreatment values (P < 0.01). Administration of simvastatin significantly lowered plasma levels of matrix metalloproteinase-9 (MMP-9) and monocyte chemoattractant protein-I [33+/ 46 and 13+/-19%, respectively (P = 0.027 and 0.020, respectively)]. Furthermore, administration of simvastatin tended to lower plasma levels of plasminogen activator inhibitor type-1 and tumor necrosis factor-alpha [by 20+/-44 and 13+/ 29%, respectively (P= 0.066 and 0.110, respectively)]. There were significant inverse correlations between pretreatment levels of MMP-9 and the degree of change in those levels after administration of simvastatin (r = -0.714, P= 0.005). However, there was no significant correlation between levels of lipoprotein and levels of MMP-9, monocyte chemoattractant protein-I, and plasminogen activator inhibitor type-1 during administration of simvastatin. CONCLUSIONS: Our current data support the hypothesis that nonlipid mechanisms elicited by administration of simvastatin contribute to the decrease in incidence of cardiovascular events and explain the early clinical benefit observed in clinical trials, independent of changes in levels of lipoprotein. PMID- 11428540 TI - Low-pressure deployment of stents: short- and long-term outcome. AB - This study investigates a strategy of low-pressure stenting with concomitant anti platelet treatment designed to prevent short- and long-term events after stenting. Ninety consecutive patients who underwent percutaneous transluminal coronary angioplasty with stenting using low-pressure stent deployment (mean 8.1 atmospheres) with concomitant anti-platelet therapy based on ticlopidine and aspirin were followed. The 30-day outcome revealed a stent thrombosis rate of 6.4%, while the 9-month major clinical event rate was 8.6%. Low-pressure stent deployment appears to confer added risk for acute or sub-acute thrombosis even when aspirin and ticlopidine are used. Conversely, low-pressure inflation is associated with excellent long-term results. PMID- 11428541 TI - A novel 32P deployable balloon system inhibits formation of neointima in porcine coronary arteries after balloon-overstretching injury. AB - BACKGROUND: Intracoronary radiation has shown the potential to reduce formation of neointima in porcine models of restenosis. OBJECTIVE: To investigate the feasibility, safety, and efficacy of a new 'deployable-balloon' device with radioisotope 32P integrated into the balloon material. METHODS: Ten swine were subjected to balloon-overstretching injury in 20 coronary arteries and were randomly allocated to receive a radiation dose of 0, 15, or 20Gy prescribed to 1 mm from the surface of the radioactive balloon material. The animals were killed 4 weeks after the procedure. Their coronaries were perfusion fixed and stained. Vessel parameters (area of intima and length of fracture) and area of thrombus area were analyzed by computer-aided histomorphometry. RESULTS: Radiation treatment with the new 32P balloon device was performed without complications. The lengts of fracture for the three groups were similar (NS). Formation of neointima after balloon injury was less in members of the radiated groups than it was in controls (area of intima/length of fracture was 0.70 +/-0.12 mm for controls, 0.08+/-0.13mm with 15 Gy, and 0.07+/-0.17 mm with 20 Gy; P < 0.001). Vessels treated with 20 Gy had a greater total area of thrombus (0.00+/-0.00 versus 0.51+/-0.98 mm2, P< 0.01). CONCLUSIONS: Intracoronary radiation therapy using a new 32P deployable-balloon system is feasible and safe. A radiation dose of 15 Gy was sufficient to inhibit neointimal response in the porcine coronary balloon-injury model. PMID- 11428542 TI - Fibrin specificity of plasminogen activators, rebound generation of thrombin, and their therapeutic implications. AB - Optimal induction of coronary thrombolysis depends in part upon the nature of the specific plasminogen activator used. The two general classes of plasminogen activators available clinically differ in a fundamental respect delineated by the term, clot selectivity. Clot selective agents are less prone to induce plasminemia and consequent occult activation of the coagulation cascade than are non-selective agents. However, under clinical conditions, all plasminogen activators result in some activation of the cascade with consequent generation of thrombin. Accordingly, optimal therapy requires the use of conjunctive anticoagulation to preclude the deleterious effects of rebound generation of thrombin, which has been well documented biochemically. The potential value of antiplatelet agents that can attenuate the positive feedback loop between activation of platelets and markedly amplified generation of thrombin in the setting of coronary thrombolysis is under active exploration. With appropriate monitoring of the efficacy of such agents in vivo it should be possible to enhance even further the benefits that can be conferred by pharmacologically induced coronary thrombolysis. PMID- 11428543 TI - Insulinoma, type 2 diabetes and plasminogen activator inhibitor type-I. PMID- 11428544 TI - Bibliography. Current world literature. PMID- 11428545 TI - The amyloid-beta peptide and its role in Alzheimer's disease. AB - Amyloid formation plays a central role in the cause and progression of Alzheimer's disease. The major component of this amyloid is the amyloid-beta (A beta) peptide, which is currently the subject of intense study. This review discusses some recent studies in the area of A beta synthesis, purification and structural analysis. Also discussed are proposed mechanisms for A beta-induced neurotoxicity and some recent advances in the development of A beta-related therapeutic strategies. PMID- 11428546 TI - A new strategy for the synthesis of cyclopeptides containing diaminoglutaric acid. AB - A new synthesis of orthogonally protected diaminoglutaric acid containing peptides using the Ugi four component condensation is presented. To demonstrate that this method is useful to replace cystine by diaminoglutaric acid in biologically interesting peptides, we built up two cyclic somatostatin analogues deriving from Sandostatin and from TT-232. A photolytically cleavable amine derivative of the nitroveratryl type is used for the Ugi four component condensation. Because of a racemic build up of the new stereocentre of the diaminoglutaric acid, and racemization of the isonitrile component, four diastereomeric peptides resulted that were separated by HPLC. The stereochemistry of the cyclopeptides could be easily and unambiguously assigned by chiral gas chromatography and a reference sample of enantiomerically pure (2S,4S) diaminoglutaric acid. PMID- 11428547 TI - Solid-Phase synthesis of a dendritic peptide related to a retinoblastoma protein fragment utilizing a combined boc- and fmoc-chemistry approach. AB - Dendritic peptides, often presented as multiple antigen peptides (MAPs), are widely used in immunological-based fields of research, although their synthesis can be extremely challenging. In this paper, a tetrameric dendritic MAP-like presentation of the retinoblastoma protein [649-654] sequence (4RB(649-654)) has been prepared using solid-phase peptide synthesis (SPPS) methods. During the synthesis of this dendritic molecule, numerous modifications to the synthetic protocols were examined. These modifications included the introduction of a combination Boc- and Fmoc-chemistry approach and also the use of 1,8 diazabicyclo[5.4.0]-undec-7-ene as a Fmoc-deprotection agent. The use in combination of Boc- and Fmoc-based synthetic strategies resulted in the production of the desired peptide molecule, 4RB(649-654), in high purity and acceptable yields following purification by reversed phase HPLC. PMID- 11428548 TI - New conformationally homogeneous beta-turn antagonists of the human B2 kinin receptor. AB - We have designed and synthesized a conformationally homogeneous series of cyclic pentapeptides of the general structure c[Pro-aa(i)-D-Tic-Oic-aa(i + 3)] which adopt a type-II' beta-turn conformation believed important for high affinity antagonism of the bradykinin (BK) B2 receptor. We incorporated D-Tic and octahydroindole-2-carboxylic acid (Oic) residues (present in known active antagonists) in a cyclic pentapeptide that would place the D-aa in the i + 1 position of the beta-turn and a proline as a bridge between the C- and N-termini sides of the turn. In positions i and i + 3 alkyl, aromatic, polar or charged amino acids could be introduced without dramatically changing the overall structure. Ten analogues were studied using 1H nuclear magnetic resonance (NMR) and evaluated for their binding affinity for the human B2 receptor. The NMR data in dimethylsulfoxide (DMSO) confirmed the structural homogeneity within the class and, on the basis of this, one representative member of the series was chosen for a detailed structure determination using NMR data in sodium dodecylsulphate (SDS) micelles and molecular dynamics calculations. Despite the structural similarity, the binding affinity of the ten analogues was strongly influenced by the nature of the side-chains in positions i and i + 3, with the doubly charged analogue 49 (pKi = 6.2) proving best. This compound may serve as the starting point for the discovery of new non-peptide bradykinin B2 receptor antagonists. PMID- 11428549 TI - Synthesis and biological evaluation of analogues of the tetrapeptide N-Acetyl-Ser Asp-Lys-Pro (AcSDKP), an inhibitor of primitive haematopoietic cell proliferation. AB - The tetrapeptide N-Acetyl-Ser-Asp-Lys-Pro (AcSDKP), an inhibitor of haematopoietic stem cell proliferation, reduces in vivo and in vitro the damage to the stem cell compartment resulting from treatment with chemotherapeutic agents or ionizing radiations. In order to provide new molecules likely to improve the myeloprotection displayed by this tetrapeptide, we have prepared a set of analogues of AcSDKP. These compounds are derived from the parent peptide by substitution or modification of the N- or of the C-terminus, or substitution of side chains. We report here that almost all investigated analogues retain the antiproliferative activity reducing in vitro the proportion of murine Colony Forming Units Granulocyte, Macrophage (CFU-GM) in S-phase and inhibiting the entry into cycle of High Proliferative Potential Colony-Forming Cells (HPP-CFC). This shows that the polar groups of Ser, Asp or Lys are critical for the expression of biological activity, but that the modification of the N- or C terminus mostly yielded compounds still retaining antiproliferative activity and devoid of toxicity. The efficacy of AcSDKP analogues in preventing in vitro the primitive haematopoietic cells from entering into cycle makes these molecules new candidates for further in vivo investigations. PMID- 11428550 TI - Transcranial Doppler in stroke. AB - Transcranial Doppler (TCD) has been extensively used in various clinical situations, and in the last two decades has established its role in the management of patients with cerebrovascular disease and stroke. Based on the Doppler principle, it uses ultrasound waves to insonate the blood vessels supplying the brain to obtain hemodynamic information. Anatomic abnormalities of vascular occlusion, stenosis and spasm can be indirectly derived. Intracranial arterial disease is an important cause of ischemic stroke and TCD can detect these with a fair amount of sensitivity and specificity. In hemodynamically significant extracranial internal carotid artery disease, TCD shows significant abnormalities in flow dynamics of the anterior circulation and abnormalities of cerebral vasomotor reactivity. A distinct advantage of TCD is the ability to monitor blood flow in a blood vessel over prolonged periods of time, which has shown microembolic signals in acute ischemic stroke, carotid artery disease, atrial fibrillation and during angiography. In acute ischemic stroke, TCD can be used to elucidate stroke mechanisms, plan and monitor treatment, and determine prognosis. In an era when stroke is increasingly being recognized as an emergency requiring immediate treatment, TCD may be capable of providing rapid information about the hemodynamic status of the cerebral circulation, within the time frame of the rather small 'therapeutic window'. TCD predicts vasospasm with a high degree of sensitivity and specificity and because of its non-invasive nature repeated assessments can be performed after subarachnoid hemorrhage. PMID- 11428551 TI - Tumour necrosis factor-alpha is increased in the cerebrospinal fluid and serum of ischaemic stroke patients and correlates with the volume of evolving brain infarct. AB - A growing body of evidence suggests the involvement of inflammatory mediators, including cytokines, in the development of ischaemic brain lesions. The aim of the present study was to investigate whether tumour necrosis factor-alpha (TNF alpha), the proinflammatory cytokine, contributes to early pathophysiological mechanisms leading to brain damage as a consequence of acute stroke. We have studied TNF-alpha levels in cerebrospinal fluid (CSF) and serum in 23 stroke patients within the first 24 hours after ischaemic stroke, confirmed by computerized tomography of the brain (CT). The control group consisted of 15 patients with the diagnosis of tension headache and neurasthenia. In stroke patients the levels of TNF-alpha both in CSF and serum were significantly higher in comparison with the control group. The positive correlation between the levels of TNF-alpha in CSF and serum of the studied patients has been observed. Furthermore, a positive correlation between both TNF-alpha levels in CSF and serum and the volume of evolving brain infarct have been shown. PMID- 11428552 TI - Liposomal antioxidants in combating ischemia-reperfusion injury in rat brain. AB - Liposome-encapsulated antioxidants have been tested in vivo to prevent oxidative attack during cerebral ischemia and reperfusion. Oxidative stress is a causal factor in the neuropathogenesis of ischemic-reperfusion injury. From the therapeutic point of view free chemical antioxidants were almost ineffective to protect cerebral tissues from those oxidative attacks. Thus an attempt has been made to prevent the oxidative damage due to the cerebral ischemic insult by the introduction of chemical antioxidants, ascorbic acid and alpha-tocopherol either encapsulated or intercalated in small unilamellar liposomes. The effectiveness of antioxidant-loaded liposomes was tested against an experimental in vivo rat model of global cerebral ischemia. Oxidative free radical attack on cerebral tissues by the ischemic insult and brief reperfusion was accounted for by the amount of diene production per unit of tissue protein. Diene production in ischemic reperfused rat brain increases almost twofold over that of the normal rats. Prevention of excess diene production has been attributed to rats when they were treated either with L-ascorbic acid-encapsulated liposomes or alpha-tocopherol intercalated liposomes 2 hours prior to the cerebral ischemic insult. Complete restriction of excess diene generation has also been achieved when a mixture of alpha-tocopherol and L-ascorbic acid-encapsulated liposomes were injected 3 hours before the ischemic infraction. PMID- 11428553 TI - Twenty-four-hour endothelin-1 secretory pattern in stroke patients. AB - Endothelin-1 (ET-1) is a potent and long-acting vasoconstrictor peptide, which may play a role in the pathophysiology of a number of diseases. Controversial data exist on its role in human ischemic stroke. In order to ascertain whether changes in ET-1 plasma levels occur in ischemic stroke, plasma ET-1 levels and mean arterial pressure were determined in 15 patients at their first ischemic cerebral infarction and in 15 control subjects, over a 24-hour period. In stroke patients, mean 24-hour plasma ET-1 levels (4.9+/-0.5 ng/L) were higher (P< 0.05) than in control subjects (3.2+/-0.3 ng/L), and correlated with the mean size of the lesion, but not with the severity score of the neurological deficit. These results support the hypothesis that ET-1 levels reflect an indicator function for the amount of damaged cerebral tissue rather than a pathophysiological role. PMID- 11428554 TI - Determination, activity and biological role of adenylosuccinate lyase in blood cells. AB - Adenylosuccinate lyase deficiency, which is associated with severe mental retardation and autistic features, was discovered in 1984. Since then this enzyme has been analyzed in many human tissues and it is now generally agreed that screening for this enzyme defect should be performed in all unexplained neurological diseases. The aim of the present study was to analyze adenylosuccinate lyase activity in blood cells by a fast simple method adaptable to screening purposes. The activity was also analyzed in B-lymphocytes from patients with B-cell chronic lymphocytic leukemia. The biological role of adenylosuccinate lyase and its importance in regulating cellular levels of AMP is discussed. PMID- 11428555 TI - Do safety practices differ between responders and non-responders to a safety questionnaire? AB - OBJECTIVE: To compare reported safety practices between responders and non responders to a safety survey. DESIGN: Cross sectional survey at baseline compared with safety practices reported at subsequent child health surveillance checks. SUBJECTS: Parents of children aged 3-12 months registered with practices participating in a controlled trial of injury prevention in primary care that did, and did not, respond to the baseline survey and who subsequently attended child health surveillance checks. RESULTS: No difference in safety practices was found between responders and non-responders to the survey at the 6-9 month check. Responders were more likely to report owning a stair gate (odds ratio (OR) 2.75, 95% confidence interval (CI) 1.82 to 4.16) and socket covers (OR 2.16, 95% CI 1.53 to 3.04) at the 12-15 month check, and owning socket covers (OR 2.19, 95% CI 1.34 to 3.61) at the 18-24 month check. Responders were more likely to report greater than the median number of safety practices at the 18 month check. CONCLUSIONS: Non-responders to a safety survey appear to be less likely to report owning several items of safety equipment than responders. Further work is needed to confirm these findings. Extrapolating the results of safety surveys to the population as a whole may lead to over estimation of safety equipment possession. PMID- 11428556 TI - Epidemiology of violent deaths in the world. AB - OBJECTIVE: This study describes epidemiologic patterns of mortality due to suicide, homicide, and war for the world in order to serve as a benchmark against which to measure future progress and to raise awareness about violence as a global public health problem. SETTING: The world and its eight major regions. METHOD: Data were derived from The Global Burden of Disease series and the US National Center for Health Statistics to estimate crude rates, age adjusted rates, sex rate ratios, and the health burden for suicide, homicide, and war related deaths for the world and its eight major regions in 1990. RESULTS: In 1990, an estimated 1,851,000 people died from violence (35.3 per 100,000) in the world. There were an estimated 786,000 suicides. Overall suicide rates ranged from 3.4 per 100,000 in Sub-Saharan Africa to 30.4 per 100,000 in China. There were an estimated 563,000 homicides. Overall homicide rates ranged from 1.0 per 100,000 in established market economies to 44.8 per 100,000 in Sub-Saharan Africa with peaks among males aged 15-24 years old, and among females aged 0-4 years old. There were an estimated 502,000 war related deaths with peaks in rates for both sexes among people aged 0-4, 15-29, and 60-69 years old. CONCLUSION: The number of violence related deaths in the world is unacceptably high. Coordinated prevention and control efforts are urgently needed. PMID- 11428557 TI - Rural population survey of behavioral and demographic risk factors for loaded firearms. AB - OBJECTIVES: In the United States, firearm deaths are almost as frequent as motor vehicle deaths. Firearm unintentional and suicide death rates are raised in rural areas. This study examines firearm prevalence and storage practices in three different types of rural households. METHODS: Adults from a stratified random sample of 983 households in a rural Iowa county were interviewed. The chi2 test of independence was used to assess association between loaded, unlocked firearms and seven behavioral and demographic risk factors. RESULTS: Nearly 67% of respondents reported firearms in their households. Nearly 7% of households had a loaded, unlocked gun. Prevalence of firearms at home was higher while prevalence of loaded, unlocked guns was lower than reported in other surveys. Prevalence of loaded, unlocked guns in farm households, 10.5%, was about twice the level in town households, 5.5% (chi2 test, p=0.033). Having taken a gun safety course was associated with more than double the prevalence of a loaded, unlocked gun, 13.5% v 5.1% (chi2 test, p=0.001). The prevalence of loaded, unlocked guns in households with a handgun, 19.3%, was four and one half times higher than in households with a long gun only, 4.2% (chi2 test, p=0.001). Households with someone with a lifetime prevalence of alcohol abuse or dependence were about twice as likely as other households, 13.0% v 6.6% (chi2 test, p=0.004), to report having loaded, unlocked firearms. CONCLUSIONS: Anyone interested in promoting safe storage of firearms in rural homes should consider these observations. PMID- 11428558 TI - Injuries among children and youth in farm households: Regional Rural Injury Study I. AB - OBJECTIVES: The purpose of this effort was to identify the incidence and consequences of both farming and non-farming related injuries and the potential risk factors for farming related injuries among children and youth, aged 0-19 years, who lived in farm households in a large region of the United States. METHODS: Data were collected from randomly selected farm households during 1990. Rates and rate ratios with 95% confidence intervals were calculated for sociodemographic and exposure variables. Multivariate analyses were conducted, using a priori and backward stepwise logistic regression models. RESULTS: Within the population of 3,939 farm households and 13,144 persons, children and youth accounted for 33%. Injury rates for farming and non-farming sources, respectively, were 1,683 and 6,980 per 100,000 persons. Animals (40%) were the primary sources of the farming operation related injuries; sports/recreation sources (61%) were associated primarily with non-farming related injuries. Of the farming and non-farming operation related injury cases, 83% and 90%, respectively, required some type of health care; moreover, 17% and 24%, respectively, were restricted from regular activities for one month or more. Through multivariate analyses, important increased rate ratios were observed for operating a tractor, working with dairy cattle, and being male. Increased rate ratios for working with beef cattle, operating a harvester, and living on a farm where there were all terrain vehicles in use, and a decreased rate ratio for living on a farm where there were sheep, appeared suggestive. CONCLUSIONS: Based on the relevant rates, injury consequences, and potential risk factors identified, injuries to children and youth on farms represent a significant problem. Future analytic studies are essential to identify more specific risk factors that can serve as a basis for development of appropriate intervention efforts. Given the population at risk, and the opportunity for intervention in this unique occupational setting, many of these injuries may be readily amenable to prevention efforts. PMID- 11428559 TI - Surveillance of hospitalized farm injuries in Canada. AB - OBJECTIVE: To provide an overview of hospital admissions for the treatment of farm injuries. METHODS: DESIGN: descriptive analysis of data from the Canadian Agricultural Injury Surveillance Program (CAISP). POPULATION: persons experiencing a farm injury requiring hospitalization, April 1991 to March 1995. Access to hospital separation data was negotiated within Canadian provinces. Individual cases were verified by medical records personnel and supplemental data describing injury circumstances were obtained. ANALYSIS: descriptive analyses characterizing farm injuries by: persons involved, mechanisms, primary diagnoses, and agents of injury. RESULTS: Data from 8/10 Canadian provinces representing 98% of the farm population were obtained. A total of 8,263 farm injuries were verified. Adults aged 60 years and older were over-represented in these injuries. Leading external causes of agricultural machinery injury included entanglements, being pinned/struck by machinery, falls, and runovers. Non-machinery causes included falls from heights, animal related trauma, and being struck/by against objects. Leading diagnoses varied by age group, but included: limb fractures/open wounds, intracranial injuries, skull fractures, and spinal/ truncal fractures. CONCLUSIONS: CAISP is a new agricultural injury surveillance program in Canada. Data from this system are actively used to inform prevention initiatives, and to indicate priorities for etiological and experimental research in the Canadian agricultural setting. PMID- 11428560 TI - Potential benefits of restrictions on the transport of teenage passengers by 16 and 17 year old drivers. AB - OBJECTIVES: The presence of passengers is associated with fatal motor vehicle crashes of teenage drivers. A restriction against newly licensed teenage drivers carrying passengers has been included in some, but not all, graduated licensing systems. The purpose of this study was to predict the net effects on all types of road users, including vehicle occupants and non-occupants, of possible prohibitions against 16-17 year old drivers carrying passengers. METHODS: Two national datasets, a census of fatal crashes and a sample of trips in the United States, were used to compute 1995 road user death rates. Potential effects of restrictions on drivers ages 16-17 carrying passengers younger than 20 were estimated, based on road user death rates and potential choices made by passengers who would have traveled with 16-17 year old drivers if there were no restrictions. RESULTS: There were 1,181 road user deaths in 1995 involving drivers ages 16-17 whose passengers were all younger than age 20. The predicted number of lives in the United States that would be saved annually ranges from 83 to 493 (corresponding to reductions of 7-42% in road user deaths) for drivers ages 16 and 17 combined. Similar percentages of reductions (8-44%) were predicted solely for 16 year old drivers. Assuming passenger restrictions would apply to all 16 year old drivers and at least one third of 17 year old drivers, an estimated 60-344 fewer deaths per year may occur if restrictions are mandated. CONCLUSIONS: Restrictions on carrying passengers younger than 20 should be considered for inclusion in graduated licensing systems. Even if fewer than half the drivers obey the restrictions, a substantial reduction in road user deaths would be expected. Further evaluation based on real world experience is needed to confirm their efficacy. PMID- 11428561 TI - Injury related infant death: the impact of race and birth weight. AB - OBJECTIVE: To examine the effect of race and birth weight independent of other sociodemographic factors on injury related infant death using national data. SETTING: Infants born in the United States to mothers who were white (non Hispanic), African American, Mexican American, and Native American. METHODS: Linked infant birth and death data from the National Center for Health Statistics for 1989-91 were analyzed to calculate unadjusted and adjusted odds ratios for death due to homicide or unintentional injury within the first year of life. In addition to maternal race and birth weight, the risk of death was adjusted for maternal age, prenatal care, maternal education, paternal education, marital status, birth order, interval since last pregnancy, smoking during pregnancy, and alcohol intake during pregnancy. RESULTS: Among 10.7 million births during 1989 91, 821 homicides and 2397 unintentional deaths were reported in infants. Relative to whites, African Americans were at highest risk for homicides (unadjusted and adjusted odds ratios = 3.6 (95% confidence interval = 3.1 to 4.2) and 1.6 (1.3 to 1.9), respectively) and Native Americans at highest risk for unintentional injuries (unadjusted and adjusted odds ratios = 3.8 (3.0 to 4.8) and 2.1 (1.7 to 2.6), respectively). After accounting for other sociodemographic factors, Mexican American infants appeared protected from injury (adjusted odds ratio = 0.7 (0.6 to 1.0) for homicides and 0.7 (0.6 to 0.8) for unintentional injuries). An inverse effect of birth weight was seen-as birth weight decreased, risk of homicides and unintentional injuries increased. After adjustment for the sociodemographic factors, very low birthweight babies were still at substantially increased risk of homicides with an odds ratio of 2.1 (1.4 to 3.1) and unintentional injuries with an odds ratio of 2.9 (2.4 to 3.7). CONCLUSIONS: Using a large national data-set, the effect of race as a risk factor for fatal infant injuries was mostly explained by birth weight and other sociodemographic factors. Preventable risk factors need to be identified for African Americans and Native Americans, in particular. Birth weight is an important independent risk factor; very low birthweight babies should be monitored for both homicide and unintentional injury. PMID- 11428562 TI - Hypothermia deaths in Jefferson County, Alabama. AB - INTRODUCTION: Some reported characteristics associated with hypothermia mortality include older age, alcohol consumption, male sex, and black race. The purpose of this paper is to present the epidemiology of hypothermia deaths in Jefferson County, Alabama. METHODS: Autopsy reports maintained by the county coroner's office were abstracted for all cases with primary or underlying causes of death listed as "hypothermia" or "exposure to cold" between January 1983 and July 1999. RESULTS: Sixty three hypothermia deaths occurred in Jefferson County during the study period. The mean age among cases was 68 years, 63.9% were male and 70% were of black race. Rates of hypothermia death were highest among black males, followed by black females, particularly blacks aged 80 years or older. Deaths occurring indoors were more common among older persons and outdoor deaths more common among younger persons. Thirty per cent of decedents tested positive for alcohol, 75% of whom were found outdoors. Nine decedents tested positive for drugs or medications. Approximately 90% of decedents were identified as having one or more chronic medical conditions. Excluding alcoholics, 52% of decedents had one or more chronic medical conditions. CONCLUSIONS: Hypothermia in Jefferson County, Alabama is a cause of death primarily affecting two distinct groups of individuals, elderly persons who develop hypothermia inside a dwelling and middle aged males who develop hypothermia out of doors and have consumed alcohol. PMID- 11428563 TI - Pupil injury risks as a function of physical and psychosocial environmental problems experienced at school. AB - OBJECTIVES: To investigate relations between physical and psychosocial environmental problems in schools, as perceived by school principals, and injuries among pupils. METHOD: Proportionate injury ratios (PIRs) were computed for 77 public sector Swedish schools (33,248 pupils), and divided into four classes based on types of environmental problems reported. Sports related injuries, injuries during recesses, and violence related injuries were considered. RESULTS: The schools reporting psychosocial problems (9.1% of schools and 7.3% of pupils) had more injuries than expected by chance than all types of injuries aggregated (PIR = 1.92; 95% confidence interval (CI) 1.64 to 2.27), and in the case of sports related injuries (PIR = 1.79; 95% CI 1.37 to 2.34) and injuries due to physical violence (PIR = 2.20; 95% CI 1.33 to 3.65). There were no significant excess risks of injuries for schools facing physical problems or a combination of physical and psychosocial problems. CONCLUSIONS: Psychosocial problems may exacerbate the risk of intentional and unintentional injuries among pupils. The results offer a reminder that school environment must be planned as part of any assessment of youth safety. PMID- 11428564 TI - Children hospitalized early and increased risk for future serious injury. AB - OBJECTIVE: To determine if infants hospitalized for any reason before 90 days of age are at increased risk for future serious injury. SETTING: Washington State. METHODS: A population based retrospective cohort study, using data from Washington State birth and death certificates linked to a statewide hospital discharge database for the years 1989 through 1997. Participants included healthy full term infants born in Washington State between 1989 and 1995. A total of 29,466 infants hospitalized <90 days of age (early hospitalization) were compared to 29,750 randomly selected infants not hospitalized early. The primary outcome was an injury resulting in hospitalization or death between 3-24 months. RESULTS: Among infants hospitalized early, 76/10,000 had a subsequent serious injury before age 2, compared with 47/ 10,000 infants without an early hospitalization (relative risk (RR) 1.6; 95% confidence interval (CI) 1.3 to 2.0). In a multivariate model including maternal age and parity, the adjusted RR for serious injury associated with early hospitalization was 1.5 (95% CI 1.2 to 1.8). Infants hospitalized early were three times as likely to be hospitalized between 3-24 months of age for intentional injury compared with infants not hospitalized early (RR 3.3; 95% CI 1.1 to 10.1). CONCLUSIONS: Infants hospitalized in the first three months of life for any reason were 50% more likely to have a subsequent serious injury compared with infants not hospitalized early and were also at increased risk of intentional injury. This identifiable group of infants might be suitable for targeted childhood injury prevention programs including those involving prenatal and postnatal visits. PMID- 11428565 TI - A geographic analysis of motor vehicle collisions with child pedestrians in Long Beach, California: comparing intersection and midblock incident locations. AB - OBJECTIVES: The purpose of this study was to use geographic information system (GIS) software to locate areas of high risk for child pedestrian-motor vehicle collisions in the city of Long Beach and to compare risk factors between midblock and intersection collisions. METHODS: Children 0-14 years of age involved in a motor vehicle versus pedestrian collision that occurred on public roadways in Long Beach, CA, between 1 January 1992 and 30 June 1995, were identified retrospectively from police reports. The GIS software program, ArcView, was used for spatial analysis and distance calculations. chi2 Tests were used to compare the distribution of the characteristics between intersection and midblock collisions. RESULTS: The average annual incident and fatality rate was 183.3/100,000 children/ year and 2.4/100,000 children/year, respectively. Children less than 5 years of age were significantly more likely to be hit at a midblock location while those aged 5-9 and 10-14 were more often hit at an intersection. Intersection collisions were more likely to occur on major arterials and local streets, and the driver to be the primary party at fault (p<0.001). While intersection incidents tended to occur further from the child's home (64.4%) the majority of midblock incidents (61.5%) occurred within 0.1 miles of the child's residence. For both midblock and intersection locations, pedestrian collisions tended to occur more frequently in those census tracts with a larger number of families per census tract-a measure of household crowding and density. CONCLUSIONS: Future studies taking into consideration traffic volume and vehicle speed would be useful to focus prevention efforts such as environmental modifications, improving police enforcement, and educational efforts targeted at parents of younger children. As GIS illustrative spatial relationships continue to improve, relationships between pedestrian collision sites and other city landmarks can advance the study of pedestrian incidents. PMID- 11428566 TI - Updating the evidence. A systematic review of what works in preventing childhood unintentional injuries: part 1. PMID- 11428567 TI - Scooter injuries to children. PMID- 11428568 TI - UNICEF's child injury league tables: a bag of mixed messages. PMID- 11428569 TI - Barriers and opportunities in reducing motor vehicle injuries. PMID- 11428570 TI - Risk compensation theory should be subject to systematic reviews of the scientific evidence. PMID- 11428571 TI - The risk compensation theory and bicycle helmets. PMID- 11428572 TI - Validation of the ICD/AIS MAP for pediatric use. AB - OBJECTIVE: To determine the performance of the ICD/AIS MAP (EJ MacKenzie et al) as a method of classifying injury severity for children. METHODS: Data on all children less than 16 years of age admitted to all designated trauma centers in Pennsylvania from January 1994 through October 1996 were obtained from the state trauma registry. The ICD/AIS MAP was used to convert all injury related ICD-9-CM diagnosis codes into abbreviated injury scale (AIS) score and injury severity score (ISS). Agreement between trauma registry AIS and ISS scores and MAP generated scores was assessed using the weighted kappa (kappaw) coefficient for ordered data and the intraclass correlation coefficient for continuous data. RESULTS: Agreement in ISS scores was excellent, both overall (intraclass correlation coefficient = 0.86, 95% confidence interval (CI) 0.84 to 0.89)), and when grouped into three levels of severity (kappaw= 0.86, 95% CI 0.85 to 0.87). Agreement in AIS scores across all body regions and ages was also excellent, (kappaw= 0.86 (95% CI 0.83 to 0.87). Agreement increased with age (kappaw= 0.78 for children <2 years; kappaw= 0.86 for older children) and varied by body region, though was excellent across all regions. CONCLUSIONS: The performance of the ICD/AIS MAP in assessing severity of pediatric injuries was equal to or better than previous assessments of its performance on primarily adult patients. Its performance was excellent across the pediatric age range and across nearly all body regions of injury. PMID- 11428573 TI - Minutes of the fourth meeting of the Working Party on Academic Education in Food Microbiology (WPAEFM) of the International Committee on Food Microbiology. PMID- 11428574 TI - Antibacterial effect of protamine in combination with EDTA and refrigeration. AB - The antimicrobial effect of protamine (clupeine) on a range of gram-positive and gram-negative foodborne pathogens and spoilage bacteria, was evaluated using an agar dilution assay and a broth dilution assay with Alamar Blue as growth indicator. Protamine was tested alone at concentrations from 0 to 10,000 microg/ml, and in combination with EDTA (0.9 mM). Assays were performed at 5 degrees C, 10 degrees C, 18 degrees C and 30 degrees C to test the effect of temperature. Minimum inhibitory concentration (MIC) values ranged from 10 microg/ml for Brochothrix thermosphacta to no inhibition at 10,000 microg/ml for bacteria such as Aeromonas hydrophila, proteolytic strains of Clostridium botulinum, Hafnia alvei and Morganella morganii. The minimum bactericidal concentrations (MBCs) were generally higher than MICs. In combination with EDTA, MICs of protamine decreased for gram-negative test strains, whereas EDTA alone inhibited gram-positive strains. The effect of assay incubation temperature was variable and not clear for most strains. Concentrations of 100-750 microg/ml protamine inhibited the five non-proteolytic C. botulinum strains, while none of the eight proteolytic strains was inhibited, indicating the possible role of proteolytic enzymes in protecting cells from protamine. Clearing zones, indicative of proteolytic activity, were observed in the opaque TSB-agarose around colonies of some but not all protamine-resistant bacteria, suggesting that this is not the only resistance mechanism. Addition of 5% (w/v) gelatin to study the effect of an increased protein concentration in the agar dilution assay showed that electrostatic interactions between protamine and the protein decreased the antimicrobial efficacy of the peptide. PMID- 11428575 TI - Microstructural factors controlling the survival of food-borne pathogens in porous media. AB - The survival of Salmonella typhimurium LT2, Escherichia coli K-12 and Pseudomonas putida in several model porous media poised at a water activity of 0.94 is shown to depend critically on the microstructure of the particulate matrix and the microscopic water distribution. The porous media were made by randomly dispersing a liquid inoculum containing ca. 10(7) cells/ml throughout the pores and interparticle spaces of packed beds of silica particles and Sephadex microspheres. The purely "microstructural stress" effects were isolated by comparison with a homogeneous liquid growth medium having the same water activity. The possibility of exploiting similar microstructural stress effects in food preservation is discussed. PMID- 11428576 TI - Characterisation of volatile compounds produced by bacteria isolated from the spoilage flora of cold-smoked salmon. AB - This study investigated the volatile compounds produced by bacteria belonging to nine different bacterial groups: Lactobacillus sake, L. farciminis, L. alimentarius, Carnobacterium piscicola, Aeromonas sp., Shewanella putrefaciens, Brochothrix thermosphacta, Photobacterium phosphoreum and Enterobacteriaceae isolated from cold-smoked salmon. Each bacterial group was represented by several strains. In addition, combinations of the groups were examined as well. Sterile blocks of cold-smoked salmon were inoculated, vacuum-packed and stored at 6 degrees C. After 40 days of storage at 6 degrees C, aerobic viable count and pH were recorded, the volatile fraction of the samples was analysed by gas chromatography-mass spectrometry (GC-MS), and spoilage was assessed by sensory evaluation. Among the 81 volatile compounds identified by GC-MS, 30 appeared to be released as a result of bacterial metabolism. Some of the effects of inoculated bacterial strains on the composition of the volatile fraction seemed to be characteristic of certain bacterial species. Sensory analysis showed relationships between bacteria, the composition of the volatile fraction and the organoleptic quality of smoked salmon. PMID- 11428577 TI - Amino acid-decarboxylase activity of bacteria isolated from fermented pork sausages. AB - The occurrence of amino acid-decarboxylase activity in 92 strains of lactic acid bacteria, coagulase-negative staphylococci, and Enterobacteriaceae isolated from Spanish fermented pork sausages was investigated. The presence of biogenic amines in a decarboxylase synthetic broth was determined by ion-pair high performance liquid chromatography with o-phtalaldehyde post-column derivatization. Among the 66 lactic acid bacteria strains tested, 21 lactobacilli (in particular, Lactobacillus curvatus) and all 16 enterococci were amine producers. Tyramine was the main amine produced by these bacteria, although they also produced phenylethylamine, tryptamine, and/or the diamines putrescine and cadaverine. None of the lactic acid bacteria produced histamine. Coagulase-negative staphylococci were found to be negative amine-producers. Aromatic monoamines, apart from histamine, were not formed by Enterobacteriaceae. This family was responsible for cadaverine and putrescine production. The results obtained for biogenic amine production by bacteria in a synthetic medium suggest that amino acid decarboxylase activity is strain dependent rather than being related to specific species. PMID- 11428578 TI - Industrial application of an antilisterial strain of Lactobacillus sakei as a protective culture and its effect on the sensory acceptability of cooked, sliced, vacuum-packaged meats. AB - The application of a protective lactic acid bacterium (LAB) during the commercial production of cooked meat products is described. The LAB, a strain of Lactobacillus sakei, was previously isolated from cooked ham and inhibited growth of Listeria monocytogenes and Escherichia coli O157:H7 in this product. L. sakei was applied to the cooked products at a concentration of 10(5)-10(6) cfu/g immediately before slicing and vacuum-packaging using a hand-operated spraying bottle. The LAB strain inhibited growth of 10(3) cfu/g of a cocktail of three rifampicin resistant mutant L. monocytogenes strains both at 8 degrees C and 4 degrees C. Consumer acceptance tests of cooked ham and of servelat sausage, a Norwegian non-fermented cooked meat sausage, showed that control and inoculated products were equally acceptable. The products were still acceptable after storage for 28 days at 4 degrees C and, after opening the packages, for a further 5 days at 4 degrees C. The findings presented here confirm that the L. sakei strain is suitable for use as a protective culture and may technically easily be implemented in the commercial production of cooked meat products. PMID- 11428580 TI - Bibilography of food microbiology. PMID- 11428579 TI - Effects of sodium lactate and other additives in a cooked ham product on sensory quality and development of a strain of Lactobacillus curvatus and Listeria monocytogenes. AB - Cooked cured ham products were produced according to a standard recipe for cooked ham with various levels of sodium lactate, sodium diacetate or buffered sodium citrate. They were compared with a reference ham product with respect to sensory quality and growth of Lactobacillus curvatus and Listeria monocytogenes. For this, a part of the products was sensory analysed directly after preparation. Another part of the cooked ham products was minced and homogeneously inoculated with L. curvatus (10(4)/g) and L. monocytogenes (10(2)/g) and filled in 60-g plastic pouches. After vacuum packaging, the pouches were stored at 4 degrees C for up to 40 days. Between the different ham compositions, only minor differences were found for appearance, internal colour, structure and firmness. The addition of 0.2% Na-diacetate had a negative effect on the odour and taste of the ham product. The addition of 2.5% to 3.3% Na-lactate inhibited the growth of L. curvatus compared to the reference, while 0.1% and 0.2% Na-diacetate did not. L. monocytogenes was best inhibited by the addition of Na-lactate but also by the addition of 0.2% Na-diacetate. On the other hand, the growth of L. monocytogenes was stimulated by the addition of 1% buffered Na-citrate. PMID- 11428581 TI - Central regulation of hepatic function by neuropeptides. AB - In addition to classical neurotransmitters, such as acetylcholine and noradrenaline, neuropeptides have been recognized as new neurotransmitters and neuromodulators. Neuropeptides are widely distributed in the central nervous system as well as in peripheral nerves, and act as neurotransmitters to regulate various physiological functions. The digestive organs are no exception, and several neuropeptides in the central nervous system are shown to act in specific brain sites and control gastrointestinal functions, such as gastric acid secretion, and gastrointestinal motility, through the autonomic nervous system. Recently, a relationship between central neuropeptides and hepatic function through the autonomic nervous system has been revealed in animal models. Thyrotropin-releasing hormone acts in the medulla, in particular in the left dorsal vagal complex, to induce stimulation of hepatic blood flow and hepatic proliferation, and protect against experimental liver injury through vagal and cholinergic pathways. Corticotropin-releasing factor injected intracisternally elicits inhibition of the hepatic blood flow and exacerbates experimental liver injury through sympathetic and noradrenergic pathways. Neuropeptide Y acts in the left dorsal vagal complex, in particular in regard to the Y1 receptor subtype, to stimulate bile secretion. Other neuropeptides such as beta-endorphin and bombesin in the brain modulate hepatic proliferation and bile secretion. Through the use of neuropeptides, new knowledge of the central and peripheral mechanisms underlying brain regulation of hepatic function will be revealed. Further studies in regard to the physiological relevance of the central action of neuropeptides on specific brain sites should be performed to unravel the underlying pathways that mediate brain-liver interaction. PMID- 11428582 TI - Incidence of the appearance of the red color sign on esophageal varices and its predictive factors: long-term observations of 359 patients with cirrhosis. AB - PURPOSE: The red color sign observed by endoscopic examination is a reliable predictive factor for variceal bleeding. The aim of this study was to calculate the incidence of the appearance of the red color sign and to evaluate its predictive factors. METHODS: Endoscopic examination was repeatedly performed in 359 consecutive patients diagnosed as having liver cirrhosis with or without esophageal varices, during a median follow-up period of 2651 days. RESULTS: The incidence of the appearance of the red color sign on esophageal varices at the end of the tenth year was compared among patients without varices (11.4%), those with small varices (45.4%), and those with mid-size varices (65.0%). The difference was significant (P < 0.0001). The number of varices (P = 0.0010), size of varices (P = 0.0064), platelet count (P = 0.0168), and alpha-fetoprotein level (P = 0.0207) were significantly correlated with the appearance of the red color sign, as estimated by the multivariate Cox hazard model. To exclude the influence of carcinogenesis, observation was stopped when hepatocellular carcinoma was discovered. Additive predictive factors with significance were: number of varices (P = 0.001), size of varices (P = 0.027), and platelet count (P = 0.0315). CONCLUSIONS: Endoscopic signs of esophageal varices and platelet count were significant predictors for the appearance of the red color sign. PMID- 11428583 TI - Effects of omeprazole and pirenzepine on enterochromaffin-like cells and parietal cells in rat stomach. AB - PURPOSE: The purpose of this study was to investigate the mechanism of the regulation of histamine synthesis in enterochromaffin-like cells, chemically and structurally, by treatment with omeprazole and pirenzepine. METHODS: The ultrastructures of enterochromaffin-like cells and parietal cells were examined in rats treated with oral omeprazole (20 mg/kg) or intraperitoneal pirenzepine (1 mg/kg) administration. Serum gastrin concentrations, mRNA levels of H+-K+-ATPase and histidine decarboxylase, and the fundic concentrations of somatostatin and histamine were determined. RESULTS: Pirenzepine treatment suppressed omeprazole induced increases in serum gastrin levels and mRNA levels of H+-K+-ATPase and histidine decarboxylase. Pirenzepine also decreased omeprazole-induced increases of histamine concentration in fundic mucosa. Pirenzepine elevated somatostatin mRNA level, previously decreased by omeprazole treatment, in fundic mucosa. In the cytoplasm of enterochromaffin-like cells, omeprazole markedly reduced the numbers of vesicles and granules, but significantly increased their diameters, whereas pirenzepine treatment changed neither of these features. The densities and diameters of both vesicles and granules produced by treatment with omeprazole and pirenzepine were between those produced by treatment with omeprazole alone and pirenzepine alone. CONCLUSIONS: Omeprazole-induced hypergastrinemia and pirenzepine-induced somatostatin synthesis play important roles not only in histamine synthesis but also in ultrastructural changes in enterochromaffin-like cells. PMID- 11428584 TI - The role of nitric oxide (NO) in the human internal anal sphincter. AB - PURPOSE: Nitric oxide (NO) has recently been shown to be a neurotransmitter in nonadrenergic noncholinergic (NANC) inhibitory nerves in the human gut. To clarify the physiological significance of NO in the human internal anal sphincter (IAS), we investigated enteric nervous responses in normal IAS muscle strips above the dentate line, obtained from patients with rectal cancer. METHODS: Normal IAS muscle strips above the dentate line, obtained from ten patients who underwent rectal amputation for low rectal cancers were used. The subjects consisted of eight men and two women, aged from 46-72 years (mean age, 54.2 years). A mechanographic technique was used to evaluate in-vitro IAS muscle responses to electrical field stimulation (EFS) of adrenergic and cholinergic nerves before and after treatment with various autonomic nerve blockers, N(G) nitro-L-arginine (L-NNA) and L-arginine. RESULTS: Excitatory nerves were mainly involved in the regulation of enteric nerve responses to EFS in the baseline condition of the study, and NANC inhibitory nerves acted on the normal IAS. L-NNA concentration-dependently inhibited the relaxation in response to EFS in the human IAS, and this inhibitory effect in the IAS was reversed by L-arginine. CONCLUSIONS: These findings suggest that NANC inhibitory nerves play important roles in regulating relaxation of the human IAS, and that NO plays an important role as a neurotransmitter in NANC inhibitory nerves of the human IAS. PMID- 11428585 TI - Clinical, virological, and pathological significance of hepatic bile duct injuries in Chinese patients with chronic hepatitis C. AB - PURPOSE: Hepatic bile duct injuries are characteristic histological findings in patients with chronic hepatitis C virus (HCV) infection. However, the pathogenesis and clinical significance of this phenomenon remain unclear. The aims of this study were to evaluate the prevalence and clinical significance of hepatic bile duct injuries in Chinese patients with chronic hepatitis C. METHODS: One hundred and seventeen Chinese patients with chronic hepatitis C were enrolled. Clinical, biochemical, immunological (serum autoantibodies and cryoglobulinemia), histological, and virological data (serum HCV RNA titer and HCV genotype) were compared between patients with and without hepatic bile duct injuries. RESULTS: Eighty-three (71%) of the 117 patients with chronic hepatitis C had hepatic bile duct injuries. Patients with hepatic bile duct injuries had a significantly higher frequency of HCV genotype 1b; a higher mean serum globulin level; significantly higher mean scores for histological periportal necro inflammation, portal inflammation, and fibrosis; and more severe portal lymphoid aggregation/follicles when compared with patients without hepatic bile duct injuries (P < 0.05, all). No significant differences in the presence of serum autoantibodies, cryoglobulinemia, mean serum HCV RNA titer, or response to interferon treatment were noted between the two groups. Multivariate logistic regression analysis showed that HCV genotype 1b infection, portal inflammation, and lymphoid aggregation/follicles were significant independent predictors associated with hepatic bile duct injuries. CONCLUSIONS: The presence of hepatic bile duct injuries in Chinese patients with chronic hepatitis C was significantly correlated with HCV genotype 1b infection, and the patients with these injuries had more severe portal inflammation and formation of lymphoid aggregates/follicles. PMID- 11428586 TI - Diagnostic value of serum markers of connective tissue turnover for predicting histological staging and grading in patients with chronic hepatitis C. AB - PURPOSE: Chronic hepatitis C is an insidiously progressive disease, in which repeated assessment of liver histology is required. Various serum fibrotic markers have now been introduced. Our present aim was to assess, by receiver operating characteristic analysis, the usefulness of serum fibrotic markers for diagnosing fibrotic staging and necroinflammatory grading in chronic hepatitis C. METHODS: Serum levels of procollagen type III N-terminal peptide (PIIINP), 7S fragment of type IV collagen (PIVNP), hyaluronan (HA), matrix metalloproteinase (MMP)-1, MMP-2, and tissue inhibitor of metalloproteinases (TIMP)-1 were measured in 169 patients with chronic hepatitis C. RESULTS: The accuracy of these tests for discriminating stages greater than F2 from stages less than F1 was superior to that for discriminating stage F3 from stages less than F2. The most useful test for predicting stages greater than F2 was the serum HA test (cutoff value, 50 ng/ml; sensitivity, 75%; specificity, 80%), and the next-most useful was the serum MMP-2 test (cutoff value, 550 ng/ml; sensitivity, 75%; specificity, 70%). The usefulness of these tests for discriminating moderate grade from grades less than mild was superior to that for discriminating grades more than mild from minimal grade. The most useful test for predicting moderate grade was the serum HA test (cutoff value, 60 ng/ml; sensitivity, 77%; specificity, 74%), and the second-most useful was the serum PIVNP test (cutoff value, 6.5 ng/ml: sensitivity, 74%; specificity, 75%). The combination of the most useful and next most useful test results increased the accuracy of the diagnosis of staging and grading. CONCLUSIONS: These serum fibrotic markers, especially the serum HA test, would be clinically useful for assessing staging and grading in patients with chronic hepatitis C. PMID- 11428587 TI - Activity of brush border membrane enzymes in proximal jejunum of portal hypertensive rats. AB - PURPOSE: Malabsorption accompanies portal hypertension, especially when associated with chronic liver disease. The development of portal hypertension is accompanied by significant alterations in the splanchnic microcirculation. In this study, the effect of extrahepatic and intrahepatic portal hypertension on brush border membrane enzymes was estimated. METHODS: Portal hypertension was induced in rats by partial portal vein ligation (PPVL) (n = 6) and common bile duct ligation (CBDL) (n = 6), and the activity of sucrase, lactase, alkaline phosphatase, and leucine aminopeptidase (LAP) in the intestinal homogenate was measured. RESULTS: Intrasplenic pulp pressure (ISPP) (in cm of saline) was found to be elevated in PPVL (21.3 +/- 1.47) and CBDL animals (21.5 +/- 1.79) as compared with findings in their respective sham-operated controls (12.74 +/- 0.86, 11.83 +/- 1.04). Only sucrase and LAP activity was significantly elevated (P < 0.05) in the PPVL group. No changes were observed in the CBDL group. CONCLUSION: Only sucrase and LAP activities were increased in PPVL rats. PMID- 11428588 TI - Adenocarcinoma complicating Barrett's esophagus: an analysis of cell proliferation. AB - In Japan, cases of Barrett's esophagus with concurrent adenocarcinoma are relatively rare. We report herein a case of long-segment Barrett's esophagus associated adenocarcinoma in a 72-year-old Japanese man. The surgical specimen showed that an ulcerating tumor, measuring 5.5 x 3.9 cm, was present in the lower esophagus adjacent to the esophagogastric junction, the background lower esophagus having an erythematous appearance. Histologically, the ulcerating tumor was a well-to-moderately differentiated tubular adenocarcinoma, with a small area of signet ring cell carcinoma invading the adventitia. In addition, the esophageal epithelium was replaced by columnar epithelium (9.5 cm in length) with multifocal dysplastic changes. Immunohistochemically, the number of Ki-67 positive cells gradually increased, moving from the normal gastric mucosa (mean Ki-67 labeling index [mKLI], 2.6%) through Barrett's epithelium (mKLI, 12.9%), low-grade dysplasia (mKLI, 16.9%), and high-grade dysplasia (mKLI. 23.7%) to invasive carcinoma, in that order, with labeling higher in the invasive tubular adenocarcinoma elements (mKLI, 40.5%) than in areas of signet ring cell carcinoma (mKLI, 20.4%). Findings in our patient suggest that increased cellular proliferation plays an integral part, in the progression of Barrett's metaplasia to adenocarcinoma. The collection of further cases for analysis will be necessary to confirm this hypothesis. PMID- 11428589 TI - Multiple hepatic infarction after transcatheter arterial infusion with SMANCS. AB - We report a patient with hepatocellular carcinoma who developed multiple hepatic infarction after transcatheter arterial infusion (TAI) with a suspension of styrene maleic acid neocarzinostatin (SMANCS) and Lipiodol (SMANCS/Lipiodol). The parameters of hepatic functional reserve were apparently decreased after the second TAI with SMANCS/Lipiodol, and the patient died of hepatic failure 103 days after the second TAI. The autopsy liver specimen revealed multiple hepatic infarctions associated with peripheral arterial stenosis or occlusion, and portal thrombosis. It is speculated that both the arterial occlusion and the portal thrombosis caused the hepatic infarction, based on a long-term insufficiency of blood supply to the hepatocytes arising from toxic arteritis caused by SMANCS/Lipiodol. PMID- 11428590 TI - Pancreatic ductal adenocarcinoma associated with Potter type III cystic disease. AB - Although polycystic liver disease (PLD) is known to be associated with autosomal dominant polycystic kidney disease, a finding of PLD with pancreatic ductal adenocarcinoma is extremely rare. We have experienced one such case of a ductal adenocarcinoma of the pancreas in a patient with Potter type III cystic disease of the liver and kidney. A 63-year-old man was admitted to our hospital because of obstructive jaundice. Six months previously, on admission to a local hospital for treatment of diabetes mellitus, he had been found to have polycystic disease of the liver and kidney. Ultrasound examination revealed dilatation of the intrahepatic bile duct and the common bile duct. Blood tests showed an elevated total bilirubin level. Abdominal computed tomography scans and magnetic resonance imaging demonstrated polycystic lesions in the liver and the bilateral kidneys. Percutaneous transhepatic cholangio-drainage was performed, and fluorography of the biliary tree revealed obstruction of the lower common bile duct, causing jaundice. This appears to be a case of independent association of pancreatic ductal adenocarcinoma with polycystic disease of the liver and kidney. The patient's sister, who also had polycystic disease of the liver and kidney, had died of squamous cell carcinoma of the tongue. Although familial associations of carcinomas with polycystic liver disease may be extremely rare, they provide a perspective for the etiology of polycystic liver disease. PMID- 11428591 TI - Sclerosing cholangitis and systemic lymphadenopathy. AB - A 48-year-old man was admitted to our hospital with jaundice and systemic lymphadenopathy. Cholangiographic findings and liver histology disclosed the presence of sclerosing cholangitis. The patient also had a marked polyclonal increase in IgG levels. The cholangiographic findings, the systemic lymphadenopathy, and the increase in IgG levels resolved completely after treatment with prednisolone. This case suggests that there is an association between sclerosing cholangitis and immunologic abnormalities, and that corticosteroid treatment is useful for this disorder. PMID- 11428592 TI - Clinical implications of the red color sign on esophageal varices. PMID- 11428593 TI - Does pirenzepine suppress the undesirable effects of a proton pump inhibitor on enterochromaffin-like cells? PMID- 11428594 TI - Potter type III cystic disease and pancreatic malignancies. PMID- 11428595 TI - Tubule-based bioartificial liver: a proposal. PMID- 11428596 TI - Cyst fluid levels of carcinoembryonic antigen, carbohydrate antigen 19-9, squamous cell carcinoma antigen, and amylase in thyroglossal duct and branchial cleft cysts. AB - Cyst fluid was aspirated from five thyroglossal duct cysts (TDCs) and four branchial cleft cysts (BCCs). The cyst fluid levels of carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 19-9, squamous cell carcinoma antigen (SCC), and amylase were measured and compared between the two types of cyst. The enzyme immunoassay technique was used for CEA and CA19-9, while SCC and amylase levels were measured by the immunoradiometric assay and nitrophenol method, respectively. Immunostaining for CEA, CA19-9, and amylase was also performed. The serum levels of these markers in both groups of patients were within the normal ranges. In contrast, their cyst fluid levels were extremely high. There was no significant difference in the cyst fluid levels of CEA and SCC between the two types of cyst; however, the cyst fluid from the TDCs showed significantly higher levels of CA19-9 and significantly lower levels of amylase compared with that from the BCCs. Immunostaining revealed expression of CA 19-9 in nearly half the columnar epithelial cells in the TDCs, but not in the squamous epithelial cells in the BCCs. CEA and amylase were not found in the epithelial cells of either type of cyst. These findings seem to reflect the difference in etiology between TDCs and BCCs. PMID- 11428597 TI - Influence of cytoimmunological state on the development of tuberculosis in heart transplant recipients. AB - We examined the relationship between the development of tuberculosis and the cytoimmunological state of orthotopic heart transplant (HTx) recipients, which is affected by immunosuppressive therapy. Tuberculosis developed in 7 (1%) of 716 HTx recipients (four men and three women, aged 33-71 years) during a 7-year period, the standardized annualizing rate being about 1370/100000 per year, which is greater than the 17.5/100000 per year in the general population of Germany. Tuberculosis developed in the early posttransplant period in four patients when they were experiencing episodes of ongoing rejection, after 2.5, 3.5, 4.0, and 9.0 months, respectively, the standardized annualizing rate being 780/100000 per year. In three of those four patients, cytoimmunological monitoring was carried out until the development of tuberculosis. The repeated administration of pulsed corticosteroid therapy followed by oral steroids reduced T-cell and CD4+ T-cell counts, which could have increased the risk of tuberculosis developing if they were exposed. The cytoimmunological state of the remaining three patients in whom tuberculosis developed late after HTx, when episodes of ongoing rejection did not exist, was similar to the preoperative state, the standardized annualizing rate being 590/100000 per year. These findings indicate that the relatively high incidence of tuberculosis in post-HTx patients could be attributable to the immunosuppressive therapy given, including steroids. PMID- 11428598 TI - Albendazole therapy in the treatment of hydatid liver disease. AB - Hydatid disease, a common health problem in Turkey as in many countries, should be included in the differential diagnosis of all patients presenting with a cystic mass in the abdominal or thoracic cavity. Previously, surgery had been the main form of treatment. But with the introduction of preoperative medication with albendazole, treating this disease medically is now growing in popularity. We report herein the findings of ten patients with two lung and 20 liver hydatid cysts ranging from 3 to 20 cm in diameter, who were successfully treated with medical therapy. Multiple organ involvement and multiple cysts in an organ are the main indications for medical treatment with albendazole, but the indications should not be limited to certain situations such as small-sized cysts, since even huge-sized cysts responded well to the therapy in this series. PMID- 11428599 TI - Long-term outcomes of immunosuppressed renal transplant recipients with malignancies. AB - This study analyzes ten cases of malignancy in a cohort of 183 renal transplant recipients, examining surgical management, postoperative immunosuppressive therapy, and long-term outcome. One of these ten patients, who had malignant lymphoma of the jejunum, died of the neoplasm, but the other nine patients did not show any signs of tumor recurrence after removal. All of these nine patients, except for one who had transplant renal cell carcinoma (RCC), received the same dose of immunosuppressive agents after surgery for the malignant disease. Seven patients were still alive at the time of this report, six of whom had good transplant renal function. The findings of this study indicate that even if immunosuppressive agents predispose to the development of cancer, it is not necessary to reduce their dose after removal of the tumor. PMID- 11428600 TI - Protective effect of a lipoxygenase inhibitor, nordihydroguaretic acid, on the ileal motor disturbances induced by Serratia marcescens endotoxin in rats. AB - This study was conducted to examine the effects of peptidoleukotrienes on the ileal contractility disturbances induced by Serratia marcescens endotoxin in rats. Thirty-two male Wistar rats were divided into four groups (n = 8 each). The first group was given only an anesthetic agent (control group); the second group was given the endotoxin (endotoxin group); the third group was given a lipoxygenase inhibitor, nordihydroguaretic acid (NDGA); and the fourth group was given NDGA 10 min before administration of the endotoxin (NDGA+endotoxin group). The isolated ileum response was recorded in each group. Normal contractile activity was seen in the control group. After the endotoxin was given. the isolated ileum did not respond to 497acetylcholine (ACh) in the endotoxin group, but the contractile results of isolated ileum to ACh were similar to the control group results in both the NDGA and endotoxin+NDGA groups. The results of this study demostrate that leukotrienes may play a role in endotoxin-induced ileal contractility disturbances, and that the lipoxygenase inhibitor, NDGA, could be useful for the treatment of ileal motility disturbances induced by endotoxin. PMID- 11428601 TI - The effects of cefephim, G-CSF, and sucralfate on bacterial translocation in experimentally induced acute pancreatitis. AB - The preventive effects of granulocyte colony-stimulating factor, cefephim, and sucralfate on bacterial translocation in experimentally induced acute pancreatitis were investigated. Forty male Wistar albino rats were used in this study. For each rat, the pancreatobiliary ductus was ligated and hence acute pancreatitis was induced. In the control group, no further procedure was performed. Meanwhile, cefephim as an antibiotic, filgrastim, which is a colony stimulating factor, and sucralfate were given to the other groups at the specified doses. To inhibit bacterial translocation by preserving the bowel barrier, sucralfate, which is known to have a cytoprotective effect on the gastrointestinal system, was used in high doses. Cefephim 30 mg/kg per day (intramuscularly) in group II, filgrastim 10 mg/kg per day (subcutaneously) in group III, and sucralfate 50 mg/kg per day by 8-F feeding tube gavage into the stomach in group IV were given. The number of bacteria translocated into the mesenteric lymph nodes, pancreas, liver, and spleen in the control group significantly increased in comparison with the other groups (P < 0.05). The average number of leukocytes (per mm3) in the control group was significantly higher than that of other groups (P < 0.0001). Regarding the average serum amylase levels, the values of all groups clearly decreased in comparison with the control group (P < 0.0001). Although in the cefephim, filgrastim, and sucralfate groups, (+) pancreatitis was generally seen, in the control group (+++) pancreatitis was detected. Bacterial translocation to the mesenteric lymph nodes and pancreas was partially prevented by filgrastim and sucralfate, and was completely prevented by cefephim. We conclude that in the management of acute pancreatitis, the use of the prophylactic antibiotics, sucralfate and filgrastim, may be advantageous. PMID- 11428602 TI - Invasive thymoma associated with lung cancer: report of a case. AB - We report herein a case of invasive thymoma simultaneously associated with lung cancer. A 64-year-old man presented with a cough and anterior chest pain, and preoperative examinations revealed an anterior mediastinal tumor as well as lung cancer. The patient underwent a total thymectomy, partial resection of the right lung, left lower lobectomy, and mediastinal lymph node dissection, followed by radiotherapy. Although it is well known that thymomas may be accompanied by nonthymic cancers, invasive thymomas occurring coincidentally with lung cancer are rarely reported in Japan. This case is very interesting in its relation to the oncogenesis of thymomas. PMID- 11428603 TI - Pulmonary carcinoid found in a patient who presented with initial symptoms of brain metastasis: report of a case. AB - Although atypical pulmonary carcinoids frequently metastasize to regional lymph nodes, they rarely metastasize to the brain. We describe herein an extremely unusual case of a 52-year-old woman who presented with the symptoms of brain metastasis as the initial manifestation of an atypical pulmonary carcinoid. After control of the multiple metastatic brain lesions had been achieved by tumorectomy and stereotactic radiosurgery, a middle lobectomy of the right lung was performed to completely resect the primary pulmonary carcinoid. This aggressive surgical approach was successful in that it prolonged the survival of the patient and enhanced her quality of life. PMID- 11428604 TI - Regression of lymph node metastases by immunotherapy using autologous breast tumor-lysate pulsed dendritic cells: report of a case. AB - A 50-year-old woman was diagnosed as having stage IV breast cancer with bilateral supraclavicular lymph node metastasis resistant to CAF therapy. She received immunotherapy using autologous tumor lysate-pulsed dendritic cells (DCs). Four cycles of DC injection into the right supraclavicular lymph nodes resulted in regression of bilateral supraclavicular lymphogenous metastasis. Histological studies revealed an accumulation of CD45+ T lymphcytes in the regressive lymph nodes. This case suggests that immunotherapy with DCs may be a safe and promising approach for the treatment of advanced breast cancer. PMID- 11428605 TI - A solitary gastric Peutz-Jeghers type polyp: report of a case. AB - Peutz-Jeghers type polyps of the stomach are rare and almost always associated with intestinal polyposis and mucocutaneous pigmentation; a condition known as Peutz-Jeghers syndrome. The case presented in this report refers to a woman found to have a large solitary Peutz-Jeghers type polyp of the stomach, with a maximal diameter of 7cm. Extended investigation did not reveal intestinal polyposis or any other sign of Peutz-Jeghers syndrome. Because of the size of the polyp, a partial gastrectomy was performed. To the best of our knowledge, there are only three other reports in the literature of a solitary Peutz-Jeghers type gastric polyp occurring in the absence of Peutz-Jeghers syndrome. This patient is scheduled to undergo a follow-up examination every 2 years to detect any sign of the development of Peutz-Jeghers syndrome or malignancies commonly associated with it. PMID- 11428606 TI - Cronkhite-Canada syndrome associated with advanced rectal cancer treated by a subtotal colectomy: report of a case. AB - A 41-year-old man with Cronkhite-Canada syndrome presented with multiple juvenile polyps with hyperplastic and adenomatous changes throughout his stomach and entire colorectum. Dysgeusia was recognized and the degree of hypoproteinemia was remarkable. A barium enema study and colonofiberscopy also revealed an advanced cancer in the rectum. Chronic hepatitis B and membranous glomerulonephritis were also present. It was difficult to design a conservative protocol using steroids for the treatment of protein-loosing enteropathy because the patient was a hepatitis B virus carrier. As a result, a subtotal colectomy while preserving the cecum with cecorectal anastomosis was performed. Pathologically, the ulcerated rectal tumor was a moderately differentiated adenocarcinoma with invasion into the muscularis propria. Most polyps showed cystically dilated glands without dysplasia or edematous stroma with inflammatory cell infiltration. A few polyps were juvenile-type polyps with adenoma components. Although no remarkable improvement was observed in the hypoproteinemia postoperatively, an alpha1 antitrypsin clearance test showed a significant decrease in protein loss from the gastrointestinal tract, which was only about one third of the loss seen preoperatively. These findings lead us to conclude that when improvement using conservative treatment can be neither obtained nor is expected, then the use of surgery should be considered when treating patients with Cronkhite-Canada syndrome. PMID- 11428607 TI - Amelanotic malignant melanoma of the rectum: report of a case. AB - We describe herein a case of amelanotic melanoma of the rectum. Macroscopically, the tumor was lacking in pigmentation and microscopically, it was found to be deficient in melanin. Moreover, the tumor cells showed positive staining for HMB 45, the antimelanoma antibody, which led to a diagnosis of amelanotic melanoma. The patient died of metastatic tumors in the liver and bone 17 months after undergoing abdominoperineal resection of the rectum and dissection of the bilateral inguinal lymph nodes. PMID- 11428608 TI - Unusual liver carcinomas with sarcomatous features: analysis of four cases. AB - We recently examined the clinicopathological and immunohistochemical features of four cases of primary hepatic carcinoma with sarcomatoid elements. Three of the four patients had associated ordinary hepatocellular carcinoma (HCC) and one had a sarcomatoid carcinoma with no apparent elements of HCC. The presenting symptoms were high fever and hypochondralgia in three patients, and right hypochondralgia without a high fever in one. The preoperative diagnoses were liver abscess in two patients, HCC in one, and cholangioma in one. Preoperative imaging showed necrotic change or abscess formation in the tumors. The sarcomatous elements showed a positive reaction to vimentin in three patients, but the ordinary HCC cells did not. Macroscopically, the tumors appeared as a single nodule with pericapsular growth. The prognoses of these patients were poor due to the early development of intrahepatic or distal metastases. We conclude that symptoms such as a high fever or hypochondralgia are characteristics of these tumors and that they may be histogenetically derived from a dedifferentiation of HCC, although no elements of HCC were found in one of our cases. PMID- 11428609 TI - Carcinoma arising in the pancreas 17 years after primary excision of a choledochal cysts: report of a case. AB - We report herein the case of a 42-year-old woman in whom a pancreatic carcinoma was found 17 years after excision of a choledochal cyst with Roux-en-Y hepaticojejunostomy. Although the incidence of biliary carcinoma associated with choledochal cyst is high, reports of cancer developing in the remaining intrapancreatic biliary tract or pancreas in patients who have previously undergone primary excision of a choledochal cyst are extremely rare. Long-term follow-up is recommended even for patients who have undergone excisional surgery for a choledochal cyst, in consideration of the possibility that cancer associated with the intrapancreatic biliary tract or pancreas may develop. PMID- 11428610 TI - Mucin-producing tumor of the pancreas associated with pyothorax: report of a case. AB - We report herein the case of a 76-year-old man for whom an invasive mucin producing tumor of the pancreas (MPTP) was successfully treated by surgery. A cystic lesion of the pancreas had been found by computed tomography (CT) 9 years earlier, 2 years following which suction drainage for left pyothorax had been carried out. A pancreatic cyst fistula to the thorax had subsequently been found during decortication for recurrent pyothorax 2 years later. Methicillin-resistant Staphylococcus aureus was detected in the pleural discharge after the thoracotomy, and thoracic fenestration was performed. A CT scan done 4 years later showed enlargement of the pancreatic cysts and a cystography revealed communication to the duodenum via the main pancreatic duct. Endoscopic retrograde cholangiopancreatography (ERCP) showed dilatation of the main pancreatic duct. The pancreatic cyst fistulated to the stomach and to the fenestrated thorax. Since MPTP was suspected from this clinical course, a distal pancreatectomy, partial gastrectomy, and omentopexy to the thorax were performed. The pathological diagnosis was intraductal papillary-mucinous tumor of the pancreas with a megacyst. While MPTP is recognized as a low-grade malignancy, some cases of invasive disease have been reported. To the best of our knowledge, this is the first case of MPTP associated with pyothorax due to fistula formation. PMID- 11428611 TI - A ruptured pancreaticoduodenal artery aneurysm associated with a splenic artery aneurysm: report of a case. AB - True pancreaticoduodenal artery (PDA) aneurysms are extremely rare. We report herein a case of a ruptured PDA aneurysm associated with a nonruptured splenic artery aneurysm which was successfully treated by surgery. A 55-year-old man was admitted to a local hospital complaining of sudden abdominal and back pain, and thereafter he was transferred to our university hospital. Abdominal computed tomography revealed retroperitoneal hematoma and an enhanced round spot suggesting a peripancreatic aneurysm. Emergency angiography showed a 20-mm-sized aneurysm in the inferior PDA and a 10-mm-sized aneurysm in the splenic artery. The patient underwent an emergency laparotomy with a diagnosis of a ruptured PDA aneurysm. After evacuating a large volume clot in the right retroperitoneal space and the peritoneal cavity, we detected an index finger-sized aneurysm with arterial bleeding in the right inferioposterior aspect of the pancreas. Hemostasis was obtained by oversewing the aneurysm and a ligation of the feeding arteries. A prophylactic splenectomy was performed for the nonruptured splenic artery aneurysm. This case indicates that emergency angiography is indispensable for both a definitive diagnosis and adequate surgical treatment of PDA aneurysms. PMID- 11428612 TI - Strangulated umbilical hernia in a child: report of a case. AB - Most umbilical hernias in children close spontaneously. Complications associated with umbilical hernias are rarely observed during follow-up. We report herein a 5 month-old girl with a strangulated umbilical hernia. Her umbilicus was hard, reddish, and irreducible. Plain radiography of the abdomen showed signs of mechanical ileus. The patient was thus diagnosed to have a strangulated umbilical hernia. A 5-cm section of the ascending colon and a 5-cm section of the terminal ileum, as well as the cecum and appendix, were congested, edematous, and erythematous, and together were enclosed by a firm hernial ring. A closure of the fascial defect and umbilicoplasty were performed. The postoperative course was uneventful. In patients with infantile umbilical hernias, strangulation may occur as the fascial defect decreases in size. PMID- 11428613 TI - Postoperative necrotizing enterocolitis following incarcerated inguinal hernia repair: report of a case. AB - The postoperative development of necrotizing enterocolitis (NEC) following major surgery in neonates has often been described. We report herein the case of an older infant in whom postoperative NEC developed following emergency repair of an incarcerated inguinal hernia. PMID- 11428614 TI - Fournier's gangrene: report of six cases. AB - Fournier's gangrene (FG) is a fatal infectious disease with necrotic fasciitis of the external genitalia. This disease persists to this day in spite of recent advances in antibiotics. Although fewer than 100 cases have been reported in Japan, we have treated six cases in the last 4 years. The patients consisted of five men and one woman, with an average age of 47.5 years. All patients received surgical treatment including incisions, aggressive debridement, drainage, irrigation, and antibiotic therapy. Two patients, who suffered from underlying diseases of diabetic nephropathy and inclusion body myositis, died. These findings confirm the fact that FG requires a prompt diagnosis and immediate surgical treatment. PMID- 11428615 TI - Angiomyofibroblastoma of the vulva: report of a case. AB - A 45-year-old woman was first seen by us 2 years after becoming aware of a slightly painful subcutaneous mass in her left vulva. The mass was 7.5 x 3.0 cm in size, well circumscribed, mobile, and rubbery. It was at first clinically considered to be a benign tumor. Microscopically, the resected mass was composed of spindle or polygonal tumor cells which were cellularly or hypocellularly arranged with perivascular accentuation in a mucoid or fibrocollagenous background. Immunohistochemically, myxoid tumor cells were positive for vimentin but not for alpha-smooth muscle actin, CD34, CD31, desmin, or S-100 protein. The tumor was diagnosed as an angiomyofibroblastoma (AMBF), based on the typical findings of histology and immunohistochemistry. There are many histological types of vulvar tumors, and establishing a preoperative diagnosis is difficult in many patients. Rapid intraoperative pathological diagnosis should be performed if possible, considering the possibility of diseases such as AMFB and aggressive angiomyxoma (AAM). When AAM is suspected, the peripheral tissues should also be resected to prevent recurrence. PMID- 11428616 TI - Laparoscopic appendectomy with the help of a wire snare. AB - This report describes the performance of a laparoscopic appendectomy (LA), a new technique using a "wire snare" without any specific complications. The snare consisted of an 18-gauge injection needle and folded fine wire, measuring 0.28 mm in diameter. LA was performed in 95 patients with acute appendicitis using a cheap and simple wire snare to catch the ligation thread of #0 silk. A good view of the appendix and mesoappendix was obtained in all cases by traction of the #0 silk. The operative time needed to perform LA using a wire snare for nongangrenous appendicitis in 77 cases overall, the first 10 cases, and the last 10 cases was 54+/-17, 64+/-5, and 33+/-4 min, respectively. Two of the 95 patients required conversion to an open appendectomy because of technical difficulties due to the formation of an inflammatory mass. There were no complications associated with insertion of the snare. The wire snare is an innovative technique and does not increase the costs significantly, and should help reduce the number of trocar sites and the operative time, while also making an LA easier to perform. PMID- 11428617 TI - The "mini-elephant trunk" technique of distal anastomosis in total arch replacement through a median sternotomy for a distal aortic arch aneurysm. AB - A median sternotomy is considered to have several advantages over a left thoracotomy as a route for the surgical treatment of a distal aortic arch aneurysm, including less pain and less lung damage. In a median sternotomy, distal anastomosis is such an important problem that we herein present a useful technique to prevent surgical bleeding from it. An invaginated 40-mm-long graft is inserted into the distal aorta, sutured to the aortic wall, unfolded, and anastomosed with a four-limbed tube graft. We applied this "mini-elephant trunk" technique to total arch replacement for both a saccural distal aneurysm, where the distal aorta was transected, and a fusiform aneurysm, where the invaginated graft was sutured by the "inclusion" method. This technique enables a greater surface contact area between the graft and the aortic wall at the distal anastomosis, thus resulting in a reduced risk of bleeding. PMID- 11428618 TI - Introduction to vesicant supplement of journal of applied toxicology. PMID- 11428619 TI - Effects of CEES on inflammatory mediators, heat shock protein 70A, histology and ultrastructure in two skin models. AB - Chemical warfare threats require the development of diverse models for the assessment of countermeasures. Human skin products, Skin2 (differentiating keratinocytes on a fibroblast-collagen matrix) and EpiDerm (differentiating keratinocytes) were exposed (2 h) to the sulfur mustard 2-chloroethyl ethyl sulfide (CEES, 1-2 mg l(-1) min(-1)) in humidified air or to humidified air alone. Tissues were evaluated histologically, ultrastructurally and for viability 22 h later; media and tissues were also analyzed for inflammatory mediators. Histology showed that CEES induced the separation of dermal and epidermal regions in Skin2 with severe damage to basal keratinocytes. Histology and electron microscopy of both products revealed condensation of nuclear chromatin, retraction of spinous processes, collapse of the tonofibrillar network and cytoplasmic vacuolization and blebbing in those cells with loss of pseudobasement membrane integrity. Exposure of Skin2 to CEES increased extracellular interleukin 1alpha (IL-1alpha), prostaglandin-E2 (PGE2) and especially IL-1 receptor antagonist (IL-1Ra) release (56,334 vs 84,614 pg ml(-1)), but decreased interleukin-6 (IL-6, 4,755 vs 351 pg ml(-1)). Exposure of EpiDerm to CEES led to unaffected extracellular and reduced intracelluar IL-1alpha (371 vs 92 pg ml( 1)). Extracellular IL-1Ra greatly increased (2,375 vs 24,875 pg ml(-1)), whereas cellular levels decreased (16,5425 vs 96,625 pg ml(-1)). Extracellular (224 vs 68 pg ml(-1)) and intracellular (485 vs 233 pg ml(-1)) soluble interleukin-1 receptor H (sIL-1RII) decreased. Prostanglandin E2 increased (1,835 vs 2,582 pg ml(-1)), whereas heat shock protein 70A (Hsp70A) remained statistically unchanged (57,000 vs 96,000 pg ml(-1)). Failure to obtain a heat shock response to CEES may contribute to the susceptibility of tissue to the alkylating agent. Consistent and marked responses of cellular and extracellular IL-1Ra to CEES suggest a potential for use as a tissue status marker and primary antiinflammatory regulator in skin. PMID- 11428620 TI - Beneficial effects of topical anti-inflammatory drugs against sulfur mustard induced ocular lesions in rabbits. AB - Ocular injuries following sulfur mustard (HD) exposure are characterized by an inflammatory response, observed as eyelid swelling, conjunctivitis, corneal oedema and cellular infiltration starting 1-4 h after exposure, depending on dose. These effects heal partially during the first 1-2 weeks after exposure, with the later appearance of neovascularization, recurrent erosions and recurrent oedema of the cornea (delayed response). We have shown previously that topically applied steroid treatment, administered after HD exposure, attenuated the extent of neovascularization, one of the characteristics of delayed ocular pathology in rabbits. The present study was designed to characterize further the initial inflammatory response and to elucidate the role of anti-inflammatory (AI) drugs as a potential therapy. Rabbit eyes were exposed to HD vapour (390 microg l(-1) for 2 min) and were treated with a topical commercial ophthalmic solution of dexamethasone or diclofenac, starting 1 h post-exposure (four times a day). Inflammation was evaluated by clinical observations, biochemical analysis of aqueous humour and by histology. Sulfur mustard exposure initiated typical clinical ocular symptoms within 4-6 h after exposure. Biochemical analysis of aqueous humour showed that protein content and prostaglandin E (PGE) increased significantly at 6 h and were still high 48 h after HD exposure. Light microscopy evaluation revealed severe damage to the cornea, characterized by epithelial denudation, oedema and cellular infiltration (mostly eosinophiles) in the stroma. Both treatments were effective in alleviating the clinical symptoms and in preventing the HD-induced increase in protein and PGE in the anterior chamber, as well as the cellular infiltration, in the corneal stroma. However, the AI treatments had no therapeutic effect on corneal erosions, and a short delay in epithelial regeneration was noted. It is concluded that AI drugs are potential candidates for the treatment of ocular lesions following HD exposure. PMID- 11428621 TI - Effects of sulfur mustard on the basal cell adhesion complex. AB - Among the most intriguing questions about sulfur mustard (di(2-chloroethyl) sulfide) is why basal cells are the primary targets of its vesicating lesions. To investigate this problem, replicate cultures of human epidermal keratinocytes (HEK) were grown from normal skin and exposed to 400 microM sulfur mustard (HD) for 5 min. Using fluorescein isothiocyanate (FITC)-conjugated antibodies, confocal laser microscopy and image analyses, we found that in early passages, sham-treated HEK maintained in a 0.15 mM Ca2+ medium continued to express keratins K5 and K14 as well as alpha6beta4-integrin. Both K5 and K14 are intermediate filaments characteristic of basal cells and linked with attachment mechanisms effecting epidermolysis bullosa simplex, a family of blistering skin diseases. Acute exposure to HD caused a statistically significant (P < 0.01) 30.74% decrease in K14 fluorescence within 1 h of exposure. Within 2 h of exposure, K14 fluorescence decreased to near-zero values. The loss in expression of K14 was progressive and occurred well before the expected appearance of in vivo blisters, which have a dose-dependent, clinical latent phase of 8-24 h. Acute exposure to HD also caused a statistically significant (P < 0.002) decrease in expression of beta4, an integrin which has been associated with junctional epidermolysis bullosa (JEB). Disruption of K14 and alpha6beta4-integrin may be early events in the HD injury pathway; however, they had no immediate or obvious effect on cell to substrate attachment. PMID- 11428622 TI - N-acetylcysteine and endothelial cell injury by sulfur mustard. AB - Understanding the underlying mechanisms of cell injury and death induced by the chemical warfare vesicant sulfur mustard (HD) will be extremely helpful in the development of effective countermeasures to this weapon of terror. We have found recently that HD induces both apoptosis and necrosis in endothelial cells (Toxicol. Appl. Pharmacol. 1996; 141: 568-583). Pretreatment of the endothelial cells for 20 h with the redox-active agent N-acetyl-L-cysteine (NAC) selectively prevented apoptotic death induced by HD. In this study, we tested the hypotheses that pretreatment with NAC acts through two different pathways to minimize endothelial injury by HD: NAC pretreatment acts via a glutathione (GSH)-dependent pathway; and NAC pretreatment acts to suppress HD-induced activation of the nuclear transcription factor NFkappaB. We used a fluorescence microscopic assay of apoptotic nuclear features to assess viability and electrophoretic mobility shift assays (EMSAs) to assess the activity of NFkappaB following exposure to HD. The cells were treated with 0-10 mM GSH for 1 h prior to and during exposure to 0 or 500 microM HD for 5-6 h. Cells were also treated with 50 mM NAC or 200 microM buthionine sulfoximine (BSO), an inhibitor of GSH synthesis, alone or in combination overnight prior to exposure to 0 or 500 microM HD for 5-6 h. Externally applied GSH up to a concentration of 5 mM had no toxic effect on the cells. Mild toxicity was associated with 10 mM GSH alone. There was a dose related enhancement of viability when 2.5 and 5 mM GSH were present during the HD exposure. Pretreatment with BSO alone had no discernible toxicity. However, pretreatment with this inhibitor of GSH synthesis potentiated the toxicity of HD. Pretreatment with 50 mM NAC, as previously reported, provided substantial protection. Combining pretreatment with both BSO and NAC eliminated the protective effect of NAC pretreatment alone on HD injury. These observations are highly suggestive that NAC enhances endothelial survival via GSH-dependent effects and confirms and extends the work of others with different models that externally supplied GSH alone may be a fairly effective countermeasure against HD injury of endothelium. We next examined the hypothesis that HD may activate the nuclear transcription factor NFkappaB by performing EMSAs with nuclear extracts of endothelial cells following exposure to 0, 250 or 500 microM HD. This demonstrated an up to 2.5-fold increase (scanning densitometry) in activation of NFkappaB binding to its consensus sequence induced by 500 microM HD after 5 h of HD exposure. Paradoxically, treatment of the endothelial cells alone with 50 mM NAC activated NFkappaB, although HD-induced activation of NFkappaB was partially suppressed by NAC at 5 h. Factor NFkappaB is an important transcription factor for a number of cytokine genes (e.g. tumor necrosis factor, TNF), which can be activated following stress in endothelial cells. Taken together, these observations suggest that the protective effects of NAC may be mediated by enhanced GSH synthesis. The increased GSH may act to scavenge HD and also prevent oxidative activation of NFkappaB. Under some conditions, NAC may act as an oxidizing agent and thus increase NFkappaB activity. The NFkappaB-dependent gene expression may be important in inducing endothelial cell death as well as in generating a local inflammatory reaction associated with the release of endothelial-derived cytokines. PMID- 11428623 TI - Treatment of sulfur mustard (HD)-induced lung injury. AB - An in vivo sulfur mustard (HD) vapor exposure model followed by bronchoalveolar lavage was developed previously in this laboratory to study biochemical indicators of HD-induced lung injury. This model was used to test two treatment compounds--niacinamide (NIA) and N-acetyl cysteine (NAC)--for their ability to ameliorate HD-induced biochemical changes. Anesthetized rats were intratracheally intubated and exposed to 0.35 mg of HD in 0.1 ml of ethanol or ethanol alone for 50 min. At the beginning of the exposure (t = 0), the rats were treated with either NIA (750 mg kg(-1)) or NAC (816 mg kg(-1)), i.p. At 24 h post-exposure, rats were euthanized and the lungs were lavaged with saline (three 5-ml washes). One milliliter of the recovered lavage fluid was analyzed for cellular components. The remaining fluid was centrifuged (10 min at 300 g) and the supernatant was assayed on a Cobas FARA clinical analyzer for lactate dehydrogenase (LDH), gamma-glutamyltransferase (GGT), albumin (ALB), total protein (TP) and glutathione peroxidase (GP). The HD alone and HD+NIA treatment caused significant increases in all of the biochemical parameters compared with control levels. The NAC treatment yielded LDH, ALB and TP values that, although elevated, were not significantly different from the control. The GP levels were significantly higher than the control but significantly lower than the HD alone levels, indicating some protection compared with the HD alone group. The GGT levels were unaffected by NAC compared with HD alone. Cytological analysis of lavage fluid showed that the percentages of neutrophils were 5.3 +/- 1.0 (mean +/ SEM) for control, 46.6 +/- 4.5 for HD, 31.4 +/- 4.7 for HD + NIA and 21.6 +/- 4.7 for HD + NAC, respectively. The neutrophil counts were significantly higher for the three HD-exposed groups vs controls; however, the NAC-treated group had neutrophil counts lower than HD alone, indicating decreased inflammatory response. These results show that NAC may be useful as a potential treatment compound for HD-induced lung injury. PMID- 11428624 TI - Role of poly(ADP-ribose) polymerase (PARP) in DNA repair in sulfur mustard exposed normal human epidermal keratinocytes (NHEK). AB - We previously reported that, in normal human epidermal keratinocytes (NHEK) cultures exposed to the alkylating compound sulfur mustard (bis-(2-chloroethyl) sulfide, HD, 0.3-1 mM), there is a rapid (< or =1 h) activation (100% above unexposed control) of the DNA repair enzyme DNA ligase I (130 kD) followed by a first-order decay (1-5 h). The DNA ligase activation is accompanied by a time dependent (0.5-4 h) and significant DNA repair. Inhibition of another putative DNA repair enzyme, poly(ADP-ribose) polymerase (PARP), by using 3-amino benzamide does not affect DNA ligase activation following HD exposure, but increases the half-life of the activated enzyme threefold. To examine the role of PARP in HD induced DNA ligase activation and subsequent DNA repair, we conducted studies using cultured keratinocytes in which the level of PARP had been selectively lowered (> or =85%) by the use of induced expression of antisense RNA. In these cells, there was no stimulation of DNA ligase up to 3 h, and a small stimulation (ca. 30% above unexposed control at 5-6 h after HD exposure. A time-course (0.5-6 h) study of DNA repair in HD-exposed PARP-deficient keratinocytes revealed a much slower rate of repair compared with HD-exposed NHEK. The results suggest an active role of PARP in DNA ligase activation and DNA repair in mammalian cells, and also indicate that modulation of PARP-mediated mechanisms may provide a useful approach in preventing HD toxicity. PMID- 11428625 TI - Treatment of skin injuries induced by sulfur mustard with calmodulin antagonists, using the pig model. AB - Sulfur mustard (HD) is a potent cutaneous vesicant that penetrates rapidly through the skin, causing prolonged injuries and leading to severe incapacitation. Although there has been long and intensive efforts to find a treatment for HD skin lesions, no effective treatment is available for HD-induced skin injuries. Recently, ointments containing calmodulin antagonists were found to be effective in preventing skin injuries induced by HD in hairless mice. The present study was designed to investigate the beneficial effects of topical treatments with calmodulin antagonists against HD skin lesions in the pig model. The pig is used as a preferred animal model for human skin in many studies, including vesicants. Neat HD, either in liquid form (0.2-1 microl droplets) or as vapour, was applied to the back skin of female pigs (a cross Large White & Landrace, 10-12 kg) for various exposure durations. Evaluation was based on quantitative analysis of the degree of erythema and area of the lesions, as well as histological evaluation. Calmodulin antagonists (10% pentamide, 1% trifluoperazine, 2% thioridazine) and anaesthetics (20% lidocaine and 3% benoxinate) were dissolved in pluronic F-127 base according to Kim et al. (Eur. J. Pharmacol. 1996; 313: 107-114) or in saline, and were applied either topically as ointments or by intradermal injection, as early as 5 min post-exposure (twice a day for at least 3 days). The results demonstrated that topically applied pluronic base ointments containing lidocaine or pentamide produce beneficial effects when applied immediately after short-term HD exposure to pig skin. PMID- 11428626 TI - Procedure for assessing myeloperoxidase and inflammatory mediator responses in hairless mouse skin. AB - A preparation procedure for making multiple inflammatory biomarker measurements from the same skin tissue was assessed. The backs of euthymic hairless mice were exposed to sulfur mustard (HD) vapor for 6 min. Animals were euthanized 24 h following exposure, dorsal skin tissue was excised and 12-mm, full-thickness biopsy punches of the exposed skin sites were taken. Specimens were snap-frozen, crushed to a powder using a biopulverizer unit, solubilized in buffer and centrifuged. Supernatant was assayed for pro-inflammatory cytokines and the acute phase reactive protein, serum amyloid P (SAP). Myeloperoxidase (MPX), which is indicative of neutrophil infiltration into the skin, was associated with the pellet fraction. Results indicate an elevation of interleukin-6, SAP and MPX in mouse skin tissue specimens 24h following HD vapor exposure. The tissue preparation procedure allows the use of a single skin specimen to make multiple inflammatory endpoint measurements requiring different preparation processes, and it will be used in subsequent studies to characterize further the inflammatory nature of HD-exposed skin tissue. PMID- 11428627 TI - Systemic administration of candidate antivesicants to protect against topically applied sulfur mustard in the mouse ear vesicant model (MEVM). AB - The mouse ear vesicant model (MEVM) provides a quantitative edema response as well as histopathological and biochemical endpoints as measurements of inflammation and tissue damage following exposure to the chemical warfare agent sulfur mustard (HD). In the MEVM, several topically applied anti-inflammatory agents provided a significant degree of protection against HD-induced edema and dermal-epidermal separation. This study evaluated the protective effects of three of these pharmacological compounds when administered systemically in the MEVM. Alzet osmotic pumps were used to deliver a subcutaneous dose of the appropriate anti-inflammatory agent, starting 24 h before exposure to sulfur mustard and continuing until 24 h post-exposure to HD. Twenty-four hours after pump implantation, 5 microl of a 195 mM (0.16 mg) solution of sulfur mustard (density = 1.27 g ml(-1); MW = 159; purity = 97.5%) in methylene chloride was applied to the inner surface of the right ear of each mouse. Sulfur mustard injury in the mouse ear was measured by both edema response (fluid accumulation) and histopathological damage (necrosis, epidermal-dermal separation). The systemic administration of hydrocortisone, indomethacin and olvanil provided a significant reduction in edema (24%, 26% and 22%, respectively) from the positive control. Compared to HD-positive controls, hydrocortisone, indomethacin and olvanil caused a significant reduction in subepidermal blisters (71%, 52% and 57%, respectively) whereas only hydrocortisone produced a significant reduction in contralateral epidermal necrosis (41%). We show here that these anti-inflammatory drugs are effective when administered systemically in the MEVM. PMID- 11428628 TI - Therapeutic approaches to dermatotoxicity by sulfur mustard. I. Modulaton of sulfur mustard-induced cutaneous injury in the mouse ear vesicant model. AB - The mouse ear edema model is recognized for its usefulness in studying skin responses and damage following exposure to chemical irritants, and for evaluating pharmacological agents against chemically induced skin injury. We recently modified the mouse ear edema model for use with sulfur mustard (HD) and used this model to study the protective effect of 33 topically applied compounds comprising five pharmaceutical strategies (anti-inflammatories, protease inhibitors, scavengers/chelators, poly(ADP-ribose) polymerase (PARP) inhibitors, calcium modulators/chelators) against HD-induced dermatotoxicity. Pharmacological modulation of HD injury in mouse ears was established by a reduction in edema or histopathology (epidermal necrosis and epidermal-dermal separation) at 24 h following topical liquid HD exposure. Ten of the 33 compounds administered as single topical pretreatments up to 2 h prior to HD challenge produced significant reductions in edema. Five of these ten also produced significant reductions in histological endpoints. Three candidates (olvanil, indomethacin, hydrocortisone) showing protection at 24 h were evaluated further for 'extended protection' at 48 and 72 h after HD challenge and showed significant modulation of edema at 48 h but not at 72 h. Olvanil also showed significant reductions in histology at 48 and 72 h. Olvanil and indomethacin were shown to reduce significantly the edema at 24 h post-exposure when administered topically 10 min after HD challenge, with olvanil additionally protecting against epidermal necrosis. These results demonstrate prophylactic and treatment effects of pharmacological agents against HD-induced skin injury in an in vivo model and support the continued use of the mouse ear vesicant model (MEVM) for evaluating medical countermeasures against HD. PMID- 11428629 TI - Sulfur mustard-induced skin burns in weanling swine evaluated clinically and histopathologically. AB - Histopathological indicators and clinical observations were used to evaluate wound severity, depth and degree of healing on days 2 and 8 for full-skin thickness sulfur-mustard (HD)-induced burns in weanling swine. Six female weanling swine were exposed for 2 h to 400 microl of HD at each of six dose sites on the hairless abdominal skin. Biopsy samples (8 mm) were taken from the periphery and from the center of the wound on day 2, and the wound was excised on day 8. Histopathological indicators evaluated were epidermal necrosis, follicular necrosis, dermal necrosis, vascular necrosis, depth of injury, ulceration (loss of epidermis), granulation tissue response, neovascularization, re epithelialization (hyperplasia) and completeness of healing. Wounds were more severe from anterior to posterior. Histopathological assessment of epidermal ulceration and necrosis of epidermis, dermis, basal epithelium, adnexal structures and subcutaneous tissue were useful indicators of wound development on day 2. Granulation tissue response (observed as early as day 8) and re epithelialization were good indicators of wound healing. Clinical evaluations were performed on day 2 prior to and after debriding, and on study day 8. Clinical observations on study day 2 were for wound size and for exudation, erythema, edema, necrosis and eschar. Clinical observations on study day 8 were for the previous parameters and for re-epithelialization, granulation and infection. Wound size and severity increased from anterior to posterior position. Size, exudation and edema were useful indicators of wound development. These histological and clinical observation parameters will be used in future experiments to compare various treatments for HD-induced burns. PMID- 11428630 TI - A cutaneous full-thickness liquid sulfur mustard burn model in weanling swine: clinical pathology and urinary excretion of thiodiglycol. AB - Sulfur mustard (bis(2-chloroethyl)sulfide, HD) is a well-known blistering chemical warfare agent. We have developed a cutaneous full-thickness HD burn model in weanling pigs for efficacy testing of candidate treatment regimens. This report addresses clinical pathology findings and the urinary excretion profile of a major HD metabolite (thiodiglycol, TDG) in this model. Six female Yorkshire pigs were exposed to HD liquid on the ventral surface for 2 h, generating six 3 cm diameter full-thickness dermal lesions per pig. Blood samples were collected throughout a 7-day observation period for hematology and serum chemistry examinations. Urine was collected in metabolism cages. Routine urinalysis was performed and the urine analyzed for TDG using gas chromatography/mass spectrometry. Examination of clinical pathology parameters revealed subtle HD related changes that are suggestive of a mild hemolytic episode. No other signs of clinically significant systemic toxicities were noted, including bone marrow suppression. Thiodiglycol was detected at the earliest time point tested (6-8 h post-exposure) at levels ranging from 0.66 to 4.98 microg ml(-1) with a mean of 2.14 microg ml(-1). Thiodiglycol concentrations were the highest for half of the animals at this earliest time point and at 24-48 h for the others. By the evening of day 3, the mean level had reached 50 ng ml(-1). Mean levels remained 10-40 ng ml(-1) for the remainder of the 7-day observation period, with the highest individual concentration noted during this period of 132 ng ml(-1). Our results are in general agreement with the TDG excretion profiles previously described for rodent models and humans. Urinary excretion of absorbed HD in our weanling pig wound healing model appears to follow the same pattern as is seen in other laboratory animals models. In general, urinary excretion of TDG appears to peak within the first 1-4 days following exposure, with detectable levels after 1 week. Relatively high urinary TDG levels may thus indicate agent exposure within the previous 96 h. Low levels significantly above natural background levels may indicate either exposure to low levels of agent or exposure that occurred more than 4 days prior to collection of the sample. PMID- 11428631 TI - Acute ocular effects of mustard gas: ultrastructural pathology and immunohistopathology of exposed rabbit cornea. AB - Whole-body exposure to sulfur mustard (HD) produces cutaneous, respiratory and ocular impairment. Of these, ocular damage causes the most immediate incapacitation. Heretofore, characterization of HD ocular toxicity has been largely limited to gross and histological observations. In the present study we explore histological, ultrastructural and immunopathological acute effects of HD ocular exposure and establish correlations with HD toxicity data already documented for dermal exposure. Anesthetized rabbits were exposed to 0.4 microl of liquid HD placed directly on the cornea. Animals were euthanized at 6, 9 and 24 h post-exposure and the eyes were enucleated and processed for histopathology, ultrastructural and immunoperoxidase study. At 6 and 9 h, the most prominent histological feature was nuclear pyknosis, necrosis and loss of polarity of corneal epithelial basal cells to the exclusion of other epithelial cells. At 24 h, all corneal epithelial cells presented degenerative changes, with the epithelium eventually detaching from the underlying basement membrane at the level of the lamina lucida. Microblisters, a characteristic HD-induced skin pathology of the basement membrane zone of animals, were absent in this corneal study. Edema, degenerating fibroblasts and inflammatory cellular infiltrates were persistent stromal responses. Immunopathological effects included changes in antigenicity of bullous pemphigoid protein, laminin, desmosonal protein, Ki67 and p53. These morphological and immunopathological effects of corneal exposure to HD appear to be largely consistent with that previously reported for dermal exposures, perhaps providing shared anatomical considerations for the development of specific HD prophylaxis and therapy. PMID- 11428632 TI - Cutaneous protease activity in the mouse ear vesicant model. AB - Tissue homogenates from mouse ear skin exposed to sulfur mustard (HD, which is a military designation and probably originated from a World War I slang term 'Hun Stuff') were assayed for serine and cysteine protease activities. Enzyme activity was measured using synthetic chromogenic thioester and fluorogenic 7-amino-4 methylcoumarin (AMC) substrates. The tissue samples were obtained from animals (n = 6) at 3, 6, 12 and 24 h post-exposure from the right ear (HD exposed), whereas control samples were obtained from the left ear (treated only with dichloromethane vehicle). The samples of naive control (left and right ear) were obtained from animals that received no HD treatment (n = 3). Elastase activity was assayed with t-butyloxycarbonyl-Ala-Ala-Ala-thiobenzylester, tryptase activity with benzyloxycarbonyl-Arg-AMC and benzyloxycarbonyl-Arg thiobenzylester, chymase activity with succinylAla-Ala-Pro-Phe-thiobenzylester and succinyl-Ala-Ala-Pro-Phe-AMC, cathepsin B activity with benzyloxycarbonyl-Arg Arg-AMC, cathepsin H activity with Arg-AMC and calpain activity with succinyl-Leu Tyr-AMC. The HD-exposed skin homogenates obtained at 12 and 24 h post-exposure had higher elastase activity (670% and 1900% increase) than control samples. For tryptase and calpain activities, only HD-exposed skin homogenates at 24h post exposure showed higher activities (220% and 170% increase) when compared to the control. No differences from control were observed for HD-exposed skin obtained at 3 and 6 h post-exposure for elastase, tryptase and calpain activities. Generally, both unexposed and HD-exposed skin had distinct cathepsin B and cathepsin H enzyme activities and small chymase activity. Enzymatic assays were also performed for other serine, cysteine and metalloproteases. These data document that proteases are involved in HD skin injury and continued assessment of proteolytic activity should be useful for identifying effective antiproteases with therapeutic use in reducing or eliminating tissue injury caused by HD cutaneous exposure. PMID- 11428633 TI - Protective effect of povidone iodine ointment against skin lesions induced by chemical and thermal stimuli. AB - Mustard gas (sulfur mustard, HD) is a powerful vesicant employed as a chemical weapon. The present study demonstrates the effect of povidone iodine (PI) ointment against skin toxicity caused by HD. Gross and histopathological examinations showed that application of PI 20 min or less following exposure to the vesicant resulted in marked skin protection. The shorter, interval between exposure and treatment, the better was the protection achieved. Povidone iodine was also effective against other mustards, such as carboxybutylchloroethyl sulfide (CBCS) and mechlorethamine. The fact that PI protected the skin against agents that cannot be oxidized, such as iodoacetic acid, divinylsulfone and cantharidine, indicated that the antidotal effect of PI was unrelated to oxidation of the nitrogen and sulfur atoms of the mustards. Furthermore, NMR spectroscopy of CBCS treated with iodine did not show oxidation of the sulfur atom. Clinical experience with patients after accidential heat burns (mostly of grade I) has shown that topical application of PI ointment immediately after the stimulus significantly reduced, and often prevented, skin lesions. Apart from being a safe and widely used disinfectant, PI ointment is recommended as an efficient protective agent against skin toxicity caused by hazardous chemicals and by heat stimuli. PMID- 11428634 TI - Diagnosis and dosimetry of exposure to sulfur mustard: development of a standard operating procedure for mass spectrometric analysis of haemoglobin adducts: exploratory research on albumin and keratin adducts. AB - Experiments were carried out to develop a standard operating procedure for analysis of sulfur mustard adducts to the N-terminal valine in haemoglobin and to explore adduct formation with albumin and keratin. In the first approach, gas chromatography-negative chemical ionization/mass spectrometry (GC-NCI/MS) of the thiohydantoin sample subsequent to the modified Edman degradation was performed using a thermodesorption/cold trap (TCT) injection technique (detection limit for in vitro exposure of human blood to sulfur mustard: 30 nM). In the second approach, the crude thiohydantoin sample was purified by solid-phase extraction procedures. In the third approach, the procedure was shortened significantly by performing the Edman degradation for 2 h at 60 degrees C. Upon exposure of human blood to various concentrations of [14C]sulfur mustard, ca. 20% was covalently bound to albumin. One of the tryptic fragments (T5 containing an alkylated cysteine (HETE-(A-L-V-L-I-A-F-A-Q-Y-L-Q-Q-C-P-F-E-D-H-V-K); MW 2536 Da) could be detected sensitively with liquid chromatography/tandem mass spectrometry analysis (detection limits: > or =15 pg absolute and 1 microM for in vitro exposure of human blood). Upon exposure of human callus (suspensions in 0.9% NaCl; 500 mg ml( 1)) to various concentrations of [14C]sulfur mustard we found 15-20% of the added radioactivity covalently bound to keratin. Upon incubation with base, 80% of the bound radioactivity was split off as [14C]thiodiglycol. This result opens the way for sensitive mass spectrometric detection of sulfur mustard exposure of skin by gas chromatography/mass spectrometry of (derivatized) thiodiglycol. PMID- 11428635 TI - Automated assay for nicotinamide adenine dinucleotide (NAD+). AB - Sulfur mustard (HD), a vesicating chemical warfare compound, has been shown to deplete the nicotinamide adenine dinucleotide (NAD+) content in several cell systems and tissues. This NAD+ depletion has been proposed as an indicator of HD exposure and can be used to evaluate potential antivesicant compounds. To examine NAD+ levels, an automated method based on the alcohol dehydrogenase cycling assay of Jacobson and Jacobson and utilizing a Cobas FARA clinical analyzer has been developed. Automation of this assay led to smaller sample volumes and more efficient use of personnel and resources. The usefulness of this automated method was tested by evaluating the protection, if any, by the topical application of vitamin D or betamethasone against HD-induced NAD+ depletion in skin punches from the hairless guinea pig. The results showed that the samples exposed to HD exhibited significant decreases in NAD+ levels when compared with controls. However, neither vitamin D nor betamethasone demonstrated protection against HD induced NAD+ depletion. In fact, betamethasone exacerbated the NAD+ depletion when compared with the HD exposed group. This assay appears to be useful for testing potential antivesicant compounds using both in vivo and in vitro exposure systems. PMID- 11428636 TI - MALDI-ToF/MS as a diagnostic tool for the confirmation of sulfur mustard exposure. AB - The continual threat of chemical and biological warfare has prompted the need for unambiguous analytical methods for the confirmation of agent exposure. In this paper, we have investigated the use of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-ToF/MS) as a diagnostic tool for this purpose. Mass spectral studies of the interaction of sulfur mustard (bis (2-chloroethyl) sulfide, HD) with hemoglobin and metallothioneine were conducted. In vitro experiments with purified proteins were performed, using both HD and chloroethylethyl sulfide (CEES), in an effort to determine the extent of alkylation and occurrence of HD cross-linking using the MALDI-ToF/MS technique. In a typical experiment, 50 ml of 5 mM HD in acetonitrile was added to an equal volume of 0.5 mM hemoglobin in deionized water followed by vortexing and incubation at room temperature. After 24 h, the samples were analyzed by MALDI ToF/MS. Mass spectral results indicated the presence of at least two distinct alkylation adducts for both HD and CEES experiments. These results demonstrate that MALDI-ToF/MS is a useful analytical technique to investigate the interaction of HD with biomolecules and may be employed potentially as a diagnostic tool for the confirmation of exposure to chemical warfare agents. PMID- 11428637 TI - Detection and measurement of sulfur mustard offgassing from the weanling pig following exposure to saturated sulfur mustard vapor. AB - Animal models are employed to investigate mechanisms of injury and to evaluate protective measures against sulfur mustard (HD) exposure. The ability to detect and quantify HD enables the researcher to follow safe procedures in handling skin samples. We designed an experimental procedure to measure HD offgassing from animal models. A Minicams--a portable gas chromatograph equipped with a flame photometric detector and on-line sorbent collection and desorption--was used to monitor the HD concentration. Confirming measurements were made using a two-step process that trapped HD on a Tenax sorbent off-line and then transferred the sample by means of an ACEM 900 to a gas chromatograph equipped with either a flame photometric detector or a mass spectrometer. Sulfur mustard offgassing data are presented from three experiments in which weanling pigs were exposed to saturated HD vapor via vapor caps containing 10 microl of HD. The HD concentration was measured in time-weighted-average (TWA) units at a specific HD application site. The current 8-h maximum exposure limit for HD is 3-ng l(-1), (1 TWA unit). The largest TWA value measured near a 3 h time point was a Minicams measurement of 0.48 TWA at 2 h and 53 min after removal of a vapor cap containing HD from a single exposure site on an animal that had 24 concurrent dorsal exposure sites. Gas chromatography/flame photometric detection and gas chromatography/mass spectrometry were used to confirm the Minicams data and to provide greater sensitivity and selectivity down to 0.1 TWA. The gas chromatography/mass spectrometry data confirmed that HD concentrations fell below 0.1 TWA in <5 h for a specific site. These measurements of HD concentrations provided information on the expeditious and safe handling of HD-exposed tissue. PMID- 11428638 TI - Comparison of cell size in sulfur mustard-induced death of keratinocytes and lymphocytes. AB - Sulfur mustard (HD) is a vesicant chemical warfare agent that directly alkylates cellular DNA and produces DNA strand breaks. To identify cellular models for in vitro screening of antivesicant compounds in DNA repair assays, we compared the mechanism of HD-induced cell death in cultured adult normal human epidermal keratinocytes (NHEK) and peripheral blood lymphocytes (PBL). One parameter that we used to distinguish apoptotic from necrotic cell death was the change in cell size due to HD. In the presence or absence of a poly(ADP-ribose) polymerase inhibitor (PARPI), cell preparations were exposed to various concentrations of HD (0.01-1.0 mM) and harvested at selected times after exposure (up to 24 h). Results from these experiments suggest that, with increasing HD concentration and time, NHEK will fragment irrespective of the presence or absence of PARPI, with cell fragmentation presumably preceded by necrosis. In the absence of PARPI, PBL size initially decreases and then remains constant over time. Previous DNA fragmentation studies indicate that both apoptosis and necrosis occur in HD exposed PBL in a time-dependent manner. In the presence of PARPI, there is a HD concentration- and time-dependent decrease in PBL size that is characteristic of apoptosis. The shift in the mechanism of HD-induced PBL death from apoptosis followed by necrosis to exclusively apoptosis in the absence and presence of PARPI, respectively, is in agreement with previous findings on HD-induced changes in membrane integrity, energy levels and DNA fragmentation. Considering that NHEK fragment early after exposure to HD concentrations that produce vesication in human skin, PBL may be a more appropriate model for use in DNA repair assays. PMID- 11428639 TI - In vitro effects of anionic sulfur compounds on the spectrophotometric properties of native DNA. AB - Several anionic sulfur compounds are recognized as efficacious pretreatments for sulfur mustard (HD) poisoning. Our intent was to see if pretreatment compounds had a direct effect on DNA, a site where HD damage is thought to occur. A modification of the method of Szinicz et al. (Arzneim.-Forsch. 1981; 31: 1,713 1,717) was used to analyze the UV/VIS spectrum (205-400 nm) (n = 6) of calf thymus DNA (10-15 x 10(3) kDa) in the absence or presence of increasing concentrations of sodium thiosulfate, sodium 2-aminoethanethiosulfonate (thiotaurine), sodium metabisulfite or sodium sulfate. All compounds produced concentration-dependent absorbance decreases primarily at 212 nm, but also at 259 nm, with the exception of sodium sulfate. For example, 8.36 x 10(-4) M sodium thiosulfate reduced the absorbance of DNA at 212 nm by >60%. The kinetics of sulfur compounds on native DNA need further study. We propose that these anionic sulfur compounds interact with DNA possibly by changing the topology of this macromolecule. Effects may be due to interactions of these sulfur compounds at higher concentrations with DNA, with resulting ligand-DNA supercoiling. This process could protect against HD intoxication, which is caused in part by the uncoiling of DNA. PMID- 11428640 TI - Organosulfur compounds as pre-exposure therapy for threat agents. AB - A series of symmetric (Ar-S-S-Ar) and unsymmetric (Ar-S-S-CH2CH2NH3+Cl-) disulfides have been prepared and evaluated as potential cyanoprotective agents. Target compounds have been prepared by known methods and/or methods developed by us specifically for this program, e.g. reaction of a thiol with 2,2' dithiobis(benzothiazole) (BT-S-S-BT) followed by reaction with a second thiol. Both 4-methoxyphenyl disulfide and 2-aminoethyl-4-methoxyphenyl disulfide hydrochloride are cyanoprotective against 2-LD50 of injected cyanide. Evaluation of both symmetric and unsymmetric related disulfides indicates that structural requirements for cyanoprotective activity are stringent and strongly suggest that protection is enzyme mediated. In addition to cyanoprotective action, initial results suggest that unsymmetric disulfides may evolve into effective antimustard agents. PMID- 11428641 TI - In vitro neurotoxic and DNA-damaging properties of nitrogen mustard. AB - Sulfur mustard and nitrogen mustard (HN2) are reported to produce neurobehavioral and neuropathological changes in animals and humans, but the mechanisms are unknown. We examined the cytotoxic properties of HN2 in cultures of dividing and post-mitotic neurons and astrocytes, which comprise the majority of cells in the central nervous system. Cultures of rat cerebellar astrocytes, post-mitotic granule cell neurons or dividing and terminally differentiated human SY5Y neuroblastoma cell cultures were treated with various concentrations of HN2 for 24 h. After treatment, culture medium was removed, the cell monolayer was incubated for 30 min with calcein-AM (green, live cells) and propidium iodide (red, dead cells) in control medium, the fluorochrome-containing medium was removed and replaced with control medium and cell density and viability were examined by fluorescence and light microscopy. Extensive cell loss (>90%) was observed in rat neuronal and SY5Y neuroblastoma cell cultures treated with 10 microM HN2, whereas cell loss was similar to controls in comparably treated astrocyte cultures. The DNA from HN2-treated cultures of rat neurons and SY5Y neuroblastoma cells was examined by high-performance liquid chromatography with electrochemical detection for the major HN2 DNA adduct N-(2-hydroxyethyl)-N[2-(7 guaninyl)ethyl]methylamine (GMOH). GMOH was detected in rat neuronal (85 fmol microg(-1) DNA) and SY5Y neuroblastoma cell cultures (46 fmol microg(-1) DNA) treated with 10 microM HN2 for 24 h, but was not detected in comparably treated astrocyte cell cultures. These findings are consistent with HN2 preferentially targeting neurons in vivo, possibly through a mechanism involving DNA damage. PMID- 11428642 TI - Calmodulin, poly(ADP-ribose)polymerase and p53 are targets for modulating the effects of sulfur mustard. AB - We describe two pathways by which the vesicating agent sulfur mustard (HD) may cause basal cell death and detachment: induction of terminal differentiation and apoptosis. Following treatment of normal human epidermal keratinocytes (NHEK) with 10 or 100 microM HD, the differentiation-specific keratin pair K1/K10 was induced and the cornified envelope precursor protein, involucrin, was cross linked by epidermal transglutaminase. Fibronectin levels were reduced in a time- and dose-dependent manner. The rapid increase in p53 and decrease in Bcl-2 levels was consistent not only with epidermal differentiation but with apoptosis as well. Further examination of biochemical markers of apoptosis following treatment of either NHEK or human papillomavirus (HPV)-immortalized keratinocytes revealed a burst of poly(ADP-ribose) synthesis, specific cleavage of poly(ADP ribose)polymerase (PARP) in vivo and in vitro into characteristic 89 and 24 kDa fragments, processing of caspase-3 into its active form and the formation of DNA ladders. The intracellular calcium chelator BAPTA suppressed the differentiation markers, whereas antisense oligonucleotides and chemical inhibitors specific for calmodulin blocked both markers of differentiation and apoptosis. Modulation of p53 levels utilizing retroviral constructs expressing the E6, E7 or E6 + E7 genes of HPV-16 revealed that HD-induced apoptosis was partially p53-dependent. Finally, immortalized fibroblasts derived from PARP -/- 'knockout mice' were exquisitely sensitive to HD-induced apoptosis. These cells became HD resistant when wild-type PARP was stably expressed in these cells. These results indicate that HD exerts its effects via calmodulin, 3 and PARP-sensitive pathways. PMID- 11428643 TI - Sulfur mustard toxicity in macrophages: effect of dexamethasone. AB - Cells from the murine macrophage-like cell line J774A.1 (J774) and cultures of primary alveolar macrophages (PAM) obtained from guinea pigs were exposed to sulfur mustard (HD, 50-200 microM) and treated with dexamethasone (2.5 microM) 10 min after HD exposure. Cell cultures were studied at 3 and 24 h after exposure by the cleavage of Thiazolyl blue reaction (MTT) reaction and crystal violet staining (viability assays), by morphological observation and by [3H]thymidine incorporation. Exposure of J774 cells to HD caused a dose-dependent decrease in viability that was evident at 24 h. Although no significant change in viability was observed at 3-4 h after HD exposure, a dose-dependent decrease in [3H]thymidine incorporation was observed. Treatment with dexamethasone caused a dose-dependent decrease in viability. However, the combined exposure to HD and dexamethasone had a synergistic effect on the decrease of cell viability. This synergistic effect is not due to a change in DNA synthesis rate because [3H]thymidine incorporation was not affected by dexamethasone. In PAM cultures, HD caused some 'activating' effect on [3H]thymidine incorporation and an increase in cell number at the lower dose (100 microM) but this was less at 200 microM. Both effects were reduced by dexamethasone treatment. We conclude that macrophages derived from different sources exhibit a different responsiveness to immunomodulators (HD and dexamethasone) and that dexamethasone can reduce the 'inflammatory' effect of HD in PAM. PMID- 11428644 TI - Novel endogenous inhibitor of sulfur mustard-stimulated protease in cultured human epidermal keratinocytes: possible application in vesicant intervention. AB - Protease stimulation at the dermal-epidermal junction may be responsible for the skin blistering (vesication) action of sulfur mustard (HD). We have purified a protease to homogeneity from cultured normal human epidermal keratinocytes (NHEK) exposed to 300 microM HD. In this report, we describe the results of our studies on purification and characterization of an endogenous inhibitor of HD-stimulated protease in NHEK. Purification to homogeneity was accomplished by chromatographic separation of the dialyzed Triton X-100-solubilized inhibitor using ion-exchange DEAE-cellulose. Analysis of the purified inhibitor by sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed one polypeptide with an apparent molecular mass of 116 kDa. Activity of the inhibitor was screened by incubating different column elute fractions with protease purified from the same cells. Preliminary results showed that the purified inhibitor effectively inhibited the protease isolated from NHEK, whereas other naturally occurring inhibitors, e.g. soybean trypsin-chymotrypsin inhibitors, elafin and aprotinin, were ineffective. Although complete characterization and regulation of this inhibitor remain to be resolved, this purification may be a major step towards developing a specific protective measure against HD-induced toxicity. PMID- 11428645 TI - Response of normal human keratinocytes to sulfur mustard: cytokine release. AB - Cytokines play a major role in both acute and chronic inflammatory processes, including those produced by sulfur mustard (2,2'-dichlorodiethyl sulfide, HD). This study describes responses of normal human epidermal keratinocytes (NHEK) to HD, defined by interleukin-1beta (IL-1beta), IL-6, IL-8 and tumor necrosis factor alpha (TNF-alpha) release. Commercially available enzyme-linked immunosorbent assay (ELISA) kits were used to measure the cytokine release in NHEK during exposure to 100 and 300 microM of HD. Exposure to 100 microM HD increased the release of cytokines. The amounts of IL-8 and TNF-alpha present in cell suspensions increased up to 59-fold and 4-fold, respectively, above control levels when NHEK were exposed to 300 microM HD. Exposure of NHEK to 300 microM HD had a highly variable effect on the release of IL-1beta, where sometimes the secretion of IL-1beta increased above baseline level and at other times it decreased in cell suspensions. Supernatants were collected from cell culture flasks 24 h after exposure of 100 and 300 microM HD and significantly increased levels of IL-6 were observed. Interleukin-6 was released in a concentration dependent manner, 3.6-fold up to 8.4-fold, respectively, in supernatant. These pro-inflammatory mediators IL-1beta, IL-8, TNF-alpha and IL-6 may play an important role in HD injury. The present findings suggest that the cytokine changes detected could be used as potential biomarkers of cutaneous vesicant injury. PMID- 11428646 TI - Reactivity of chloroethyl sulfides in the presence of a chlorinated prophylactic: a kinetic study by EPR/spin trapping and NMR techniques. AB - This study reports the kinetic reaction of a chlorinated glycoluril, 1,3,4,6 tetrachloro-7, 8-diphenyl-2,5-diimino glycoluril, also known as S-330, with butyl 2-chloroethyl sulfide (half-sulfur mustard, H-MG) and bis-(2-chloroethyl) sulfide (sulfur mustard, HD) using electron paramagnetic resonance (EPR)/spin trapping and nuclear magnetic resonance (NMR) techniques. Both H-MG and HD are highly reactive in water and are capable of alkylating a variety of critical target molecules. It is well known that compounds containing reactive chlorine are useful neutralizers of HD and other vesicating agents. Organic compounds containing a chloroamide group are generally preferred. Currently, the reactive mechanism of this chlorinated glycoluril with these chloroethyl sulfides has not been documented. The kinetic experiments were performed by adding the monofunctional sulfur mustard (H-MG) directly to the spin trap agent alpha-phenyl N-tert-butylnitrone (PBN, pH 7.1). The intensity of the EPR spectra obtained from the resulting spin adduct (hyperfine coupling constants aN = 1.45 mT and abetaH = 0.225 mT) was sensitive to the rate at which the spin adduct was formed. Different concentrations of the chloroamide were added to the reaction mixtures of PBN and H-MG. The EPR spectra of separate identical reaction mixtures were recorded with the spectrometer set for kinetic experiments. The rate constant determined by EPR was 1.78 +/- 0.14 x 10(7) M(-1)s(-1). It was found that S-330 reacts 55 times faster than PBN. The results obtained for S-330 by EPR indicate that S-330 is an efficient scavenger of H-MG. Furthermore, a 13C-NMR chemical shift of 0.903 +/- 0.002 ppm was observed for the Cl-N-C-N-Cl carbon in S-330 after exposure to HD (1 mM). In addition, the decay of 13C-NMR resonance at 91.7 ppm chemical shift was observed in the presence of HD. The 13C-NMR data showed that the formation of the ethylene sulfonium ion usually found in the case of HD was not observed in the presence of S-330. PMID- 11428647 TI - Inflammatory cytokine response in sulfur mustard-exposed mouse skin. AB - Assessment of anti-inflammatory therapies against sulfur-mustard (bis(2 chloroethyl)sulfide, HD)-induced skin injury has mainly relied on qualitative histopathological evaluation. Development of quantifiable inflammatory biomarkers using fast and reliable molecular methods is needed for screening anti inflammatory drugs against HD injury. In this study, we used two different HD exposure models to determine the in vivo cutaneous response of the inflammatory cytokines interleukin-6 (IL-6), IL-1alpha, IL-1beta and tumor necrosis factor alpha (TNF-alpha), in order to identify a suitable inflammatory biomarker common to both models. In the first model, the backs of hairless mice were exposed to HD vapor (1.4 g m(-3)) or sham controls for 6 min using an occluded vapor cup technique. In the second model, right ears of CD1 mice were exposed to a solution (5.0 microl of 195 mM) of HD (0.16 mg) in dichloromethane (CH2Cl2) whereas left ears received only CH2Cl2 (vehicle control). Sulfur-mustard-induced skin inflammation was assessed in skin punch specimens collected at time points up to 24 h post-exposure. Edema was determined by measuring tissue weight, and cytokine content was measured by enzyme immunosorbent assay. Characterized by an increase in edema and IL-6, HD provoked a cutaneous inflammatory response in both models beginning at 6 h post-exposure and continuing to 24 h. An increase in IL-1alpha was observed only in the hairless mouse model, also beginning at 6 h post exposure and continuing to 24 h. No IL-1beta or TNF-alpha response was observed at any time point in either exposure model. These data document the in vivo production of cutaneous IL-6, a distinct inflammatory biomarker, in two different HD exposure models. We conclude that IL-6 should be a useful in vivo biomarker for evaluating anti-inflammatory drugs against HD-induced skin injury. PMID- 11428648 TI - Exposure of human epidermal keratinocyte cell cultures to sulfur mustard promotes binding of complement C1q: implications for toxicity and medical countermeasures. AB - Sulfur mustard (HD)-increased proteolytic activity, HD-enhanced expression of Fc receptor (FcR) on human epidermal keratinocytes (HEK) and associated inflammatory responses may contribute to HD pathology. Like the FcR, the first component of the classical complement (C') cascade, C1q, binds to the Fc region of antibody to mediate inflammatory responses. Complement C1q binds specifically to the C1q receptor (C1qR) on the blebs of apoptotic human keratinocytes and is proposed as a cell surface marker for apoptosis. Assays by fluorescent antibodies demonstrated significantly enhanced binding of C1q to HEK cell cultures exposed to HD. The cell populations of HEK that showed enhanced C1q binding also demonstrated an intermediate uptake of propidium iodide that was greater than in viable unexposed cells but less than in dead cells. The HD-enhanced C1q binding was concentration-dependent, negative by flow cytometry or weakly positive by digital scanning microscopy at 100 microM and positive by both methods at 300 microM. Binding of C1q was also time-dependent, weakly positive at 8 h, and positive at 16 and 24 h after HD exposure. The HD-increased C1qR that binds C1q to the surface of HEK might be a contributing mechanism or a marker for the inflammation and vesication associated with HD exposure. PMID- 11428649 TI - Sulfur mustard induces apoptosis and necrosis in SCL II cells in vitro. AB - Sulfur mustard (bis(2-chloroethyl) sulfide, HD) is an alkylating agent causing erythema and blistering with a latency of several hours after skin exposure. In the present in vitro study the influence of HD (1 microM-1 mM for 30 min or 4 h) on the viability and growth of SCL II cells was investigated. No significant differences in cytotoxicity were observed as assessed by formazan formation from XTT tetrazolium salt at 24, 48 and 72 h after exposure. Sulfur mustard concentrations of >500 microM were associated with an increasing portion of apoptotic cells without change in necrosis rate as assessed by nuclear morphology and gel electrophoresis of the DNA. The ATP levels were not affected up to 6 h after HD exposure (< or =1 mM). Twelve hours later, ATP depletion was observed at HD concentrations of >500 microM. Colony-forming ability was impaired at concentrations of <1 microM. Cell growth studies in comparison with nuclear morphology indicated late apoptotic death predominating at lower concentrations of HD. In summary, the data show that HD may inhibit cell growth already at concentrations where viability parameters and cell metabolism are not yet affected. PMID- 11428650 TI - Intervention of sulfur mustard toxicity by downregulation of cell proliferation and metabolic rates. AB - Metabolically active and proliferating basal cells in the skin are most sensitive to the potent skin blistering chemical warfare compound HD (bis-(2-chloroethyl) sulfide). We previously described a Ca2+-dependent mechanism of HD (0.3-1 mM) toxicity that was inhibited by the cell-permeant Ca2+ chelator BAPTA AM (1,2 bis(O-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester). We describe some cellular effects of BAPTA AM that suggest a mechanism for its protective action. Monolayer log-phase normal human epidermal keratinocytes were incubated (37 degrees C) first in keratinocyte growth medium (KGM) containing BAPTA AM (10-40 microM) for 30 min and then in KGM alone overnight prior to evaluation. The BAPTA AM inhibited cell growth in a concentration-dependent manner with some cellular degeneration above 30 microM (light microscopy). At 20 30 microM, BAPTA AM also inhibited cellular metabolic processes, as evidenced by a lower incorporation of [3H]-thymidine (DNA synthesis, 54 +/- 5%), [3H]-uridine (RNA synthesis, 29 +/- 6%) and [14C]-valine (protein synthesis, 12 +/- 2%) as well as a lower protein content per culture (30 +/- 3%) compared with corresponding untreated controls. However, 20-30 microM BAPTA AM did not cause any demonstrable cytopathology based on morphological (electron microscopy) as well as biochemical (lactate dehydrogenase release, an indicator of cell viability loss) criteria, indicating a lack of acute toxicity. These results suggest that a mechanism of protection by BAPTA AM against HD may be via decreasing some metabolic, and therefore proliferative, rates. PMID- 11428651 TI - Poly(ADP-ribose) polymerase inhibitors regulate the mechanism of sulfur mustard initiated cell death in human lymphocytes. AB - Sulfur mustard (HD) produces slow-healing skin lesions that contain large, tight fluid-filled blisters. These lesions are the result of severe damage to areas of the body exposed to HD and require extensive medical care before complete recovery is achieved. Converting the mechanism of HD-initiated cell death from an inflammatory oncosis (homicide) to benign apoptosis (assisted suicide) may reduce the extent of cellular damage and the time required for healing. HD-exposed human lymphocytes lose cellular function, membrane integrity and viability, and suffer degradation of their nuclear components. The treatment of HD-exposed cells with poly(ADP-ribose) polymerase inhibitors prevents or alters the HD-initiated loss of cell viability, membrane integrity, cellular metabolic constituent (NAD) and cellular energy (ATP), while initiating alterations in nuclear constituents. It is hoped that by preventing or altering these HD-initiated changes we can limit the extent of the injury, decrease the time required for repair and reduce the loss of performance suffered by exposed individuals. The use of poly(ADP-ribose) polymerase inhibitors to assist in initiating apoptosis in affected cells should help to achieve these objectives while preventing the chance of further disease development later in the exposed individuals. PMID- 11428652 TI - Non-invasive assessment of the effects of iontophoresis on human skin in-vivo. AB - The stratum corneum (SC), the outermost layer of the skin, presents a formidable barrier to transdermal drug delivery. As a result, different strategies have been developed to enhance drug transport into and through skin. Iontophoresis involves the application of a small electrical current which drives molecules across the skin and controls relatively well the rate of delivery. Although the technique has been widely investigated in-vitro, the evaluation of skin integrity in-vivo after iontophoresis is absolutely necessary for the future clinical application of this approach. This paper reviews the non-invasive biophysical techniques which have been used to assess the effects of current application on human skin in-vivo. Specifically, transepidermal water loss, infrared spectroscopy, impedance spectroscopy and skin blood flow measurements are discussed. After first presenting the basic principles of these methods, their application to the determination of SC barrier function and skin integrity is addressed, and the criteria for selecting the most appropriate approach are considered. PMID- 11428653 TI - Triton-X-100-modified polymer and microspheres for reversal of multidrug resistance. AB - Triton X-100 is a non-ionic detergent capable of reversing multidrug resistance (MDR) due to its interaction with cell membranes. However, it interacts with cells in a non-specific way, causing cytotoxicity. This work aimed to develop polymeric chemosensitizers that possess the ability to reverse MDR and lower toxic side effects. When being delivered to tumours, the polymeric chemosensitizers may also have longer retention times in tumours than the free detergent. Triton-X-100-immobilized dextran microspheres (T-MS) and inulin (T-IN) were prepared and characterized. Their cytotoxicity against multidrug-resistant Chinese hamster ovary cells (CH(R)C5) was compared with that of free Triton X-100 solutions. The in-vitro effect of the products on 3H-vinblastine accumulation by CH(R)C5 cells was determined. Both T-MS and T-IN showed a marked decrease in the cytotoxicity, as compared with free Triton solutions at equivalent concentrations. Drug accumulation by CH(R)C5 cells was increased over two fold in the presence of T-MS or T-IN. These results suggest that polymeric drug carriers with MDR-reversing capability and lower cytotoxicity may be prepared by immobilization of chemosensitizers. PMID- 11428654 TI - Novel approach to improve permeation of ondansetron across shed snake skin as a model membrane. AB - The purpose of this study was to investigate the feasibility of transdermal drug delivery of ondansetron, an antagonist of the 5-HT3 receptor, used for the treatment of chemotherapy-induced emesis. The permeability of ondansetron from an aqueous suspension through shed snake skin as a model membrane was very low and in order to improve it, several enhancers were tested. Ethanol increased the flux at a concentration of 40% or more. The solubility of ondansetron also increased as the ethanol concentration increased. The permeability coefficient increased after pretreatment of the shed snake skin with Azone, oleic acid or lauryl alcohol. Further improvement of the permeability was observed when ethanol was combined with other enhancers and was maximum for the combination of ethanol and oleic acid. Oleic acid dramatically increased the partition of ondansetron to n hexane and shed snake skin. Oleic acid may enhance the permeation of ondansetron in two ways: by a direct effect on the stratum corneum or via counterion formation of an ion-pair. The maximum flux obtained from the combination of ethanol and other enhancers seems to be high enough to obtain a therapeutic effect. PMID- 11428655 TI - Nefiracetam metabolism by human liver microsomes: role of cytochrome P450 3A4 and cytochrome P450 1A2 in 5-hydroxynefiracetam formation. AB - An in-vitro study was conducted to investigate the metabolism of nefiracetam in human liver microsomes and to identify the enzymes responsible for the metabolism. Nefiracetam was hydroxylated by human liver microsomes to 5 hydroxynefiracetam (5-OHN). Eadie-Hofstee plots for the formation of 5-OHN suggested substrate activation. The kinetic parameters, apparent Km, Vmax, and Hill coefficient, for the formation of 5-OHN by pooled human liver microsomes were 4012 microM, 2.66 nmol min(-1) (mg protein)(-1), and 1.65, respectively. The formation of 5-OHN was significantly correlated with cytochrome P450 (CYP)3A4 mediated testosterone 6beta-hydroxylase activity and dextromethorphan N demethylase activity. The 5-OHN formation was inhibited (94%) by antibody to human CYP3A4/5. The 5-OHN formation was also inhibited by the CYP3A4 inhibitors ketoconazole and troleandomycin, but not significantly inhibited by several other P450 inhibitors. The microsomes containing cDNA-expressed CYP3A4 formed 5-OHN with sigmoidal kinetics. CYP3A5-containing microsomes did not form 5-OHN. These results indicated that CYP3A, most likely CYP3A4, was the major isozyme responsible for the formation of 5-OHN in human liver microsomes. CYP1A2 and CYP2C19 microsomes were also capable of forming 5-OHN. However, the contribution of CYP1A2 was considered to be relatively minor compared with that of CYP3A4, and the contribution of CYP2C19 was assumed to be negligible, based on the result of the immunoinhibition study and taking into account both the turnover rate by each isozyme and the relative abundance of each isozyme in human liver. We conclude that on average the formation of 5-OHN, the major metabolite of nefiracetam, is principally mediated by CYP3A4 with a relatively minor contribution by CYP1A2. PMID- 11428656 TI - Effect of experimental renal dysfunction on bioavailability of ajmaline in rats. AB - The effect of renal dysfunction on the bioavailability of ajmaline has been investigated in rats, where experimental renal dysfunction was induced by subcutaneous injection of uranyl nitrate (10 mg kg(-1)). Renal dysfunction did not cause any change in the blood ajmaline concentration after intravenous administration (2 mg kg(-1)), but it increased the blood ajmaline concentration by approximately 2.8-fold after intraduodenal administration (10 mg kg(-1)). The availability of ajmaline in control rats was 16.7%, whereas the availability was increased to 41.1% in rats with renal dysfunction. The unbound fraction in the blood and the metabolic activity in the liver, was assessed with the 10000-g supernatant fraction and with isolated hepatocytes, respectively. The values were found to be similar in both groups. The blood concentration following intraportal infusion was only slightly increased in rats with renal dysfunction, but the hepatic first-pass extraction was infusion rate-dependent and saturable. The initial absorption rate of ajmaline from the small intestine in rats with renal dysfunction was significantly greater compared with control rats. These results indicated that the increased availability of ajmaline in renal dysfunction was mainly a result of partially saturated extraction in the liver, which was caused by an increased absorption rate in the intestine and non-linear extraction in the liver. PMID- 11428657 TI - Contradistinction between doxorubicin and epirubicin: in-vitro interaction with blood components. AB - The molecular structure and anti-tumour activity of doxorubicin and epirubicin are similar. However, the incidence of their cardiotoxicity occurs at different cumulative dose concentrations. The purpose of this study was to investigate the in-vitro interaction of these two drugs with different blood components, namely intact erythrocytes, haemoglobin and erythrocyte ghosts. Plasma protein binding was also evaluated. The intended goal was to identify the most relevant samples among total blood, plasma or blood cells for pharmacokinetic analysis. The methodology involved the incubation of each of the blood components (the intact erythrocytes, erythrocyte ghosts, haemoglobin and plasma proteins) at physiological pH and temperature with different concentrations of each drug, followed by measurement by HPLC and fluorometry at excitation and emission wavelengths of 480 and 580 nm, respectively. The results indicated that the binding of doxorubicin and epirubicin to plasma proteins, erythrocyte ghosts and intact erythrocytes was essentially the same. However, the binding of both compounds to intact erythrocytes was significantly different from erythrocyte ghosts, which indicates that haemoglobin plays an important role in the binding to and uptake by erythrocytes. The isotherms of binding to haemoglobin revealed that the maximum binding of doxorubicin was approximately 0.42 microg mg(-1) haemoglobin; for epirubicin this value was ten times greater than for doxorubicin. The Scatchard plot of binding of both drugs to haemoglobin exhibited two distinct binding sites for each drug. The constant of association of high affinity and low capacity binding sites was significantly greater for epirubicin, whereas the constant of association of low affinity and high capacity binding sites was significantly higher for doxorubicin. The number of high affinity binding sites per mg of haemoglobin was estimated to be 0.072 for doxorubcin and 0.030 for epirubicin. The number of low affinity binding sites was significantly greater for epirubicin (1.963) than for doxorubicin (0.305). Since the combined number of binding sites for epirubicin was more than doxorubicin, and the total uptake by erythrocytes remained the same for both drugs, it was concluded that epirubicin, being a more lipophilic compound, may diffuse more freely into the cells. Therefore, it binds more to haemoglobin, whereas doxorubicin remains more adsorbed on the surface of the cells due to its self-association property. It was concluded that the interaction of both drugs with erythrocytes, although it appears to be similar, is significantly different due to the interaction with haemoglobin. The difference in this interaction is expected to influence the disposition of both drugs in-vivo. PMID- 11428658 TI - Inhibition by carbapenem antibiotic imipenem of intestinal absorption of valproic acid in rats. AB - The concomitant use of carbapenem antibiotics with valproic acid has been prohibited because panipenem induced a decrease in plasma concentration of valproic acid in epileptic patients during valproic acid therapy. To clarify the possible mechanism of the carbapenem-valproic acid interaction, we investigated the effect of imipenem on the pharmacokinetic behaviour of valproic acid in rats. Co-administration of imipenem (30 mg kg(-1), i.v.) induced a decrease in the peak plasma concentration of valproic acid after oral administration. However, the imipenem-induced decrease in plasma concentrations of valproic acid was not observed within 60 min after intravenous injection of valproic acid. By utilizing in-situ vascular and luminal perfused small intestine, it was confirmed that absorption of valproic acid from the luminal to the vascular perfusate was decreased in the presence of imipenem (0.5 mM) in the vascular perfusate. The everted gut sac method was used to determine the effect of imipenem on active transport of valproic acid. The accumulation of valproic acid on the serosal side of the intestinal sac against the concentration gradient was reduced by lactic acid that inhibits the carrier-mediated transport of valproic acid across the intestinal brush-border membrane. However, imipenem did not affect the active transport of valproic acid. Therefore, the inhibition by imipenem of valproic acid absorption may be caused by a mechanism different from that of lactic acid. In conclusion, imipenem inhibits the intestinal absorption of valproic acid, which contributes to the decrease in plasma concentration of valproic acid after oral administration. PMID- 11428659 TI - Synthesis of potential aldose reductase inhibitors based on minimal pharmacophore requirements. AB - A series of 17 compounds were synthesized based on the premise that the minimal pharmacophore for aldose reductase inhibition requires the presence of both an aryl group and polar group connected by a linking structure. Three groups of compounds were synthesized, the first possessing an aniline-4-(2'-6' methylbenzothiazole) or 2-aminobenzothiazole group as the aryl group, the second possessing a 2-naphthyl as the aryl group and the third possessing either a 4-(2 phenylthiazole) or 2-(5-2'-nitrophenylfuran) as the aryl group. In all three of these groups the carboxylate or its methyl ester are linked to the aryl group through various lengths of methylene carbons and amide or cinnamide groups. Optimal activity was observed when the carboxylic group was separated from the aryl group by a linking structure of five atoms in length. Both a double bond and an amide moiety are well tolerated in the linking structure. PMID- 11428660 TI - Synthesis, chemical and enzymatic hydrolysis, and bioavailability evaluation in rabbits of metronidazole amino acid ester prodrugs with enhanced water solubility. AB - A series of amino acid esters (3a-e) have been synthesized and evaluated as potential prodrugs of metronidazole with the aim of improving aqueous solubility and therapeutic efficacy. The aqueous solubility and the lipophilicity (expressed as the log P value) of metronidazole and its esters were investigated. In general the prodrugs revealed enhanced water solubility compared with metronidazole. N,N diethylglycinate hydrochloride (3a) and 4-ethylpiperazinoacetate (3e) derivatives displayed higher aqueous solubility, which exceeded that of the parent drug by factors of approximately 140 and 100, respectively. All the esters revealed lower log P values than metronidazole except for the 4-phenylpiperazinoacetate derivative (3f), which was 6.5-times more lipophilic than metronidazole. The hydrolysis kinetics of the esters were studied in aqueous phosphate buffer (pH 7.4) and 80% human plasma at 37 degrees C. In all cases the hydrolysis followed pseudo-first-order kinetics and resulted in a quantitative reversion to metronidazole as evidenced by HPLC analysis. The prodrugs exhibited adequate chemical stability (half-life, t1/2, 4-16 h) in aqueous phosphate solution of pH 7.4. In 80% human plasma they were hydrolysed within a few minutesto metronidazole. The esters 3d (methylpiperazinoacetate derivative) and 3f were exempted since their t1/2 values were approximately 2.5 and 8.5 h, respectively. A comparative pH-rate profile study of N,N-diethylglycinate hydrochloride (3a) and 4-ethyl-piperazinoacetate (3e) derivatives in aqueous buffer solution over the pH range 2.2-10 was investigated. The results indicated that 3a showed marked stability at pH 2-6 followed by accelerated hydrolysis at pH 7.4. The basic ester 3e was found to be less stable at lower pH values but exhibited comparative stability at physiological pH. Moreover, in-vivo experiments in rabbits revealed a higher metronidazole plasma level with sustained release characteristics within the prodrug-treated animals (10- and 2.5-fold) as compared with the parent drug treated group. In conclusion, the designed amino acid esters 3a and 3c-e might be considered as good candidates for water-soluble prodrug forms of metronidazole. PMID- 11428661 TI - Comparison of chemical components and antioxidants capacity of different Echinacea species. AB - Alcoholic extracts of the roots and leaves of three Echinacea species (E. purpurea, E. angustifolia and E. pallida) were analysed for the presence of characteristic chemicals by HPLC directly coupled to ultraviolet absorbance and electrospray mass spectrometric detectors. The method permitted rapid characterization and tentative identification of a large number of caffeoyl conjugates and alkamides in all the samples investigated. The roots of the three species differed markedly in their contents of characteristic compounds. Cichoric acid and verbascoside predominated in extracts of E. purpurea root whereas cynarine and dodeca-2E,4E,8Z,10Z/E-tetraenoic acid isobutylamide were the major chemicals characteristic of E. angustifolia root extracts. Echinacoside and 6-O caffeoylechinacoside predominated in extracts of E. pallida roots. Characteristic alkamides were also examined by electrospray tandem mass spectrometry (MS/MS) and these compounds provided characteristic fragmentation patterns. Extracts of the roots and leaves of all three species were found to have antioxidant properties in a free radical scavenging assay and in a lipid peroxidation assay. PMID- 11428662 TI - Pharmacological investigation on nigrescigenin-a cardenolide from Parquetina nigrescens (Afzel.) Bullock: comparative studies on cardiotonic effects of Parquetina nigrescens, g-strophanthin and noradrenaline in guinea-pig isolated atria. AB - The cardiotonic and catecholamine-like effects of Parquetina nigrescens extract induced contractile force of guinea-pig left and right atria were investigated in vitro. Isometric contractions were recorded. P. nigrescens extract, 5-150 microg mL(-1), increased the force of contraction dose dependently in electrically driven left atria. The concentration of P. nigrescens extract producing 50% of the maximal effect (EC50 value) was 7.5 microg mL(-1). The positive inotropic response differed from that of g-strophanthin by its high rate of onset and its complete reversibility upon removal of the extract from the incubation medium. In spontaneously beating right atrial muscle, P. nigrescens extract increased the rate of contractions. Its positive chronotropic and inotropic effects were partly antagonized by propranolol and atenolol indicating the presence of an adrenergic acting principle in P. nigrescens extract. In contrast, the inotropic response to P. nigrescens extract could not be completely suppressed by beta-blocking agents, suggesting that the force of contraction is not only increased by a sympathomimetic ingredient of P. nigrescens extract but also by the cardenolides known to be present in P. nigrescens. PMID- 11428663 TI - Anti-inflammatory activity of abietic acid, a diterpene isolated from Pimenta racemosa var. grissea. AB - The anti-inflammatory activity of abietic acid, a diterpene isolated from Pimenta racemosa var. grissea (Myrtaceae), was evaluated in-vivo and in-vitro. This compound significantly inhibited rat paw oedema induced by carrageenan in a time- and dose-dependent manner, and mouse ear oedema induced by 12-O tetradecanoylphorbol acetate, after oral or topical administration. The inhibition of myeloperoxidase enzyme showed that its topical activity was influenced by neutrophil infiltration into the inflamed tissues (ears). In addition, the effect of abietic acid on some macrophage functions was analysed in vitro. Non-toxic concentrations of abietic acid inhibited prostaglandin E2 (PGE2) production in lipopolysaccharide-treated macrophages, whereas nitrite, tumour necrosis factor alpha and interleukin-1beta production were only weakly affected by this diterpene. PGE2 production from A23187-stimulated macrophages was only inhibited at high doses (100 microM) and it failed to modify leukotriene C4 production. These results indicate that abietic acid exerts in-vivo anti inflammatory activity after oral or topical administration and has partial ability to prevent the production of some inflammatory mediators. PMID- 11428664 TI - Expression and function of P-glycoprotein in rats with carbon tetrachloride induced acute hepatic failure. AB - Acute hepatic failure was induced experimentally in rats by intraperitoneal injection of 2.5 mL kg(-1) carbon tetrachloride (CCl4), and the effects on the expression and function of P-glycoprotein in the liver, kidney and brain were evaluated. The CCl4 injection significantly increased the indicators of hepatic function (glutamate oxaloacetate transaminase, glutamate pyruvate transaminase), but not of renal function (blood urea nitrogen, glomerular filtration rate). In rats with acute hepatic failure, the hepatic P-glycoprotein concentration increased 1.5-fold and the ATP concentration decreased to approximately 40% that in control rats. In contrast, P-glycoprotein concentrations in the kidney and brain and ATP concentrations in the kidney remained unchanged. The in-vivo P glycoprotein function in these tissues was suppressed as evaluated by biliary and renal secretory clearances and brain distribution of rhodamine 123, a P glycoprotein substrate. These findings suggest that factors other than P glycoprotein concentration are involved in the systemic suppression of P glycoprotein function in diseased rats. In Caco-2 cells, plasma collected from CCl4-treated rats exhibited a greater inhibitory effect on P-glycoprotein mediated transport of rhodamine 123 than that from control rats, suggesting the accumulation of an endogenous P-glycoprotein substrate/inhibitor in the plasma of diseased rats. In fact, the plasma concentration of corticosterone, an endogenous P-glycoprotein substrate, increased 2-fold in CCl4-treated rats compared with control rats. It was demonstrated that P-glycoprotein function is systemically suppressed in rats with CCl4-induced acute hepatic failure, not only in the target organ (liver), but also in other organs (kidney and brain), although the P glycoprotein concentration remained unchanged in the kidney and brain, and increased in the liver. In the systemic suppression of the P-glycoprotein function in the diseased state, the alteration of plasma concentrations or components of endogenous P-glycoprotein-related compounds, such as corticosterone, would likely be involved. PMID- 11428665 TI - Arachidonic acid and its metabolites are involved in the expression of neocortical spike-and-wave spindling episodes in DBA/2J mice. AB - This work was undertaken to study the effects of dexamethasone, indometacin and mifepristone plus dexamethasone on the neocortical spike-and-wave spindling episodes (S&W) in the electrocorticogram of DBA/2J mice. Our data indicate that both dexamethasone and indometacin (1, 10, 100 microg kg(-1), i.p.) reduced the S&W of DBA/2J mice. This effect appeared 30 min after drug administration and lasted for the duration of the recording period (240 min). Mifepristone, a glucocorticoid receptor antagonist (1, 10, 100 microg kg(-1), i.p.), injected 2 h before dexamethasone, totally blocked the steroid effect. These results indicate that both dexamethasone and indometacin significantly reduce the S&W of DBA/2J mice, suggesting a possible involvement of arachidonic acid and its metabolites in the development of brain excitability. PMID- 11428666 TI - A tryptamine analog with high affinity to the heart tissues is a potential antiarrhythmic agent. AB - A novel tryptamine analog, 1-methyl-3-[N-(3-indolyl)ethyl]carbamoyl-1,4 dihydropyridine (T-CDS) was synthesized and converted into a stable, solid complex with 2-hydroxypropyl-beta-cyclodextrin. An aqueous solution of the complex was given intravenously to dogs and the concentration of T-CDS and its corresponding quaternary (T-Q+) forms were monitored in the blood for 50 min. The effect of the drug on vital heart parameters was monitored throughout the studies. At the end of the experiment the dogs were sacrificed and the concentration of the quaternary pyridinium form (T-Q+) was determined in the different heart tissues, as well as in the kidney, liver, lung, brain, urine and cerebrospinal fluid. The compound was found to be selectively bound to the heart muscles and showed different concentrations in different heart tissues. The T-Q+ concentrations were much higher in the heart after administration of the dihydro form (T-CDS), than after administering T-Q+ directly. The compound was found to be active on certain vital signs of the cardiovascular system and could be an effective and safe antiarrhythmic agent. PMID- 11428667 TI - Azo-containing urethane analogues for colonic drug delivery: synthesis, characterization and in-vitro evaluation. AB - A urethane-based analogue containing an azo aromatic linkage in the backbone was synthesized for use in colon-specific delivery systems by reacting toluene-2,6 diisocyanate with a mixture of an aromatic azo diol, (bis-4-hydroxyphenyl)-4,4' diazobiphenyl, poly(ethylene glycol) (Mn = 4000; number-average molecular weight) and 1,2-propanediol (propylene glycol). The resultant compounds (UR-1 and UR-2) were characterized by IR spectroscopy, 1H NMR spectroscopy, DSC studies, X-ray diffraction studies and molecular weight determination by gel permeation chromatography. The compounds exhibited low molecular weight, lacked film-forming properties and crystallinity in the structure. An in-vitro bacterial degradation test to demonstrate the susceptibility of azo bond to bacterial enzymes was performed using media inoculated with lactobacillus culture. The results indicated degradation of films by azoreductase. In-vitro permeation of 5 aminosalicylic acid was studied in control and lactobacilli-treated films. The permeability of the lactobacilli treated films was significantly increased suggesting the potential of these compounds for application in colonic targeting. PMID- 11428668 TI - Evaluation of absorbability of centpropazine in rats: in-situ and in-vivo appraoches. AB - Intestinal absorption of centpropazine was studied in rats by both in-situ (closed-loop method) and in-vivo (portal-venous difference) approaches. The drug was found to be well absorbed from solution in in-situ studies. However, the results obtained in-vivo suggested that very low amounts of drug reach the portal circulation after oral dosing. This could imply extensive binding to the mucosa or metabolism in the intestinal wall. The presence of higher amounts of metabolites in the portal vein compared with the inferior vena cava samples signal their formation in the gastrointestinal tract or enterohepatic recirculation. These findings will be useful in incorporating suitable structural and formulation modifications for enhancing the bioavailability of centpropazine and its analogues. PMID- 11428669 TI - Acute cardio-respiratory effects in rats of PS4alpha, an antineoplastic peptidoglycan from Mycobacterium vaccae. AB - PS4alpha is a high molecular weight peptidoglycan extracted from Mycobacterium vaccae, which has demonstrated considerable antineoplastic activity in-vivo without apparent toxicity. Available fortesting in only small quantities, a sensitive in-vivo method for measuring pulse and breathing rates in cannulated rats was applied to this compound at doses of 5, 50 and 500 microg kg(-1). Various parameters (mean arterial pressure, maximum transpulmonary pressure, compliance, heart rate, minute volume, respiratory rate and tidal volume) were followed for up to 1 h and demonstrated no significant deviation in the baseline values obtained before injection. This compound at doses up to 500 microg kg(-1) had no apparent acute toxicity in rats, but chronic effects at this and higher doses have to be determined by more conventional toxicological methods before proceeding to evaluate PS4alpha as an antineoplastic agent. PMID- 11428670 TI - Effect of fudosteine, a new cysteine derivative, on mucociliary transport. AB - We examined the effect of fudosteine ((-)-(R)-2-amino-3-(3 hydroxypropylthio)propionic acid) on the mucociliary transport (MCT) rate in quails. The MCT rate was estimated by ash transport velocity on the tracheal mucosa of quails. Fudosteine (500 mg kg(-1), p.o.) did not affect the normal MCT rate. However, topical application of fudosteine to the tracheal mucosa dose dependently protected the impairment of the MCT rate caused by exposure to cigarette smoke. The results suggest that fudosteine may participate in the defence mechanism in the respiratory tract against irritant gases. PMID- 11428671 TI - Potentiation of nerve growth factor-induced elongation of neurites by gelsemiol and 9-hydroxysemperoside aglucone in PC12D cells. AB - In PC12D cells, nerve growth factor (NGF) increased the proportion of neurite bearing cells and made neurites longer. A methanol extract of Verbena littoralis H. B. K. collected in Paraguay only slightly potentiated the proportion of PC12D cells with neurites but markedly increased the length of neurites in the presence of NGF (2 ng mL(-1)). The methanol extract was partitioned between ethyl acetate and water followed by further extraction of water fraction with n-butanol. The potentiating activity of NGF-action was observed in the ethyl acetate and n butanol fractions. The n-butanol fraction was separated by silica gel chromatography, monitoring the NGF-potentiating activity to give gelsemiol and 9 hydroxysemperoside aglucone (9-OHSA). Neither compound (30-300 microM) exhibited neurite-inducing activity alone. Gelsemiol (100-300 microM) markedly enhanced an increase in the proportion of neurite-bearing cells and an extension of the neurite length in the presence of NGF (2 ng mL(-1)). Interestingly, in the presence of NGF (2 ng mL(-1)), 9-OHSA (100-300 microM) enhanced the elongation of neurites without affecting the increase in the proportion of cells with neurites. These results suggested that gelsemiol and 9-OHSA were major active components of V. littoralis in the NGF-potentiating action. It was possible that the mechanism of neurite elongation by NGF was different from that of the increase in the proportion of neurite-bearing cells, and that 9-OHSA selectively affected the neurite elongation mechanism. PMID- 11428672 TI - Enhanced cytotoxicity of doxorubicin encapsulated in liposomes with reconstituted Sendai F-proteins. AB - Sendai F-virosomes, a novel type of liposome with reconstituted Sendai F proteins, have been tested as a delivery system for various bioactive materials. However, encapsulation limitations and difficulties in controlling their constituents were drawbacks for further application to therapeutic purposes. We have tried to control virosomal constituents and have enhanced drug encapsulation efficiency into the virosomes. In vitro cytotoxicity of doxorubicin encapsulated in the F-virosomes were compared with free doxorubicin and doxorubicin in conventional liposomes. The F-virosomes were spontaneously prepared by detergent dialysis, a reconstitution process of Sendai F-proteins into liposomes. The reconstitution density of F-proteins affected the vesicle size of virosomes prepared by detergent dialysis; the larger amount of F-proteins made a smaller size of virosomes. There was little variation of size with time at physiological conditions, whilst the vesicle size of virosomes increased at acidic storage conditions (pH 5.5). Doxorubicin encapsulated in the F-virosomes exhibited a lower IC50 against B16BL6 mouse melanoma cells and Chang human hepatocarcinoma cells than that in conventional liposomes. The F-virosomes also exhibited higher cellular uptake than conventional liposomes. Addition of dioleoylphophatidylethanolamine, a fusogenic phospholipid, into the F-virosome further increased the cellular uptake as well as in vitro cytotoxicity. These types of virosome formulations can be clinically applicable as versatile vesicles for the efficient delivery of various therapeutic drugs, including genetic materials. PMID- 11428673 TI - Evaluation of protein release from chitosan-alginate microcapsules produced using external or internal gelation. AB - A series of experiments was undertaken to evaluate the diffusion of a model protein, i.e. bovine serum albumin (BSA), from chitosan-alginate microcapsules produced using either internal or external gelation. Diffusion of BSA was quantified during the microcapsule manufacture processes (gelation, washing, rinsing) and during incubation in conditions simulating the pH encountered during the gastric and intestinal phases of digestion. Encapsulation of an acid phosphmonoesterase permitted in situ protein localization, providing evidence to explain results obtained with BSA. There was significantly greater protein loss from internally versus externally-gelled chitosan-alginate microcapsules during the manufacture process (37.6% versus 4.7%, respectively). Similar trends were observed during 24 h incubation in 0.1 N hydrochloric acid. Increasing alginate concentration from 2-4% (w:v) did not significantly reduce losses from internally gelled microcapsules. Addition of 0.25 M NaCl to the gelling medium significantly increased protein diffusing during microcapsule manufacture and acid incubation from externally gelled microcapsules. In situ protein localization revealed a higher level of protein near the surface of the microcapsules of externally gelled microcapsules versus internal gelation. The above data indicate that externally-gelled microcapsules are inhomogeneous with a higher concentration of alginate near the microcapsule surface, thus reducing the porosity of the resulting microcapsules. These results suggest that the porous nature of internally-gelled chitosan-alginate microcapsules may result in low encapsulation efficiency, depending on the nature of the product being encapsulated. PMID- 11428674 TI - In vitro evaluation and intra-articular administration of biodegradable microspheres containing naproxen sodium. AB - The dispersion of non-steroidal antiinflammatory drugs (NSAIDs) into biodegradable polymeric matrices have been accepted as a good approach for obtaining a therapeutic effect in a predetermined period of time meanwhile minimizing the side effects of NSAIDs. In the present study, it was aimed to prepare Naproxen Sodium (NS), (a NSAID) loaded microsphere formulation using natural Bovine Serum Albumin (BSA) and synthetic biodegradable polymers such as poly(lactide-co-glycolic acid) (PLGA) (50:50 MW 34,000 and 88,000 Da) for intra articular administration, and to study the retention of the drug at the site of injection in the knee joint. NS incorporated microspheres were evaluated in vitro for particle size (the mean particle size; for BSA microspheres, 10.0 +/- 0.3 microm, for PLGA microspheres, 9.0 +/- 0.2 and 5.0 +/- 0.1 microm for MW 34,000 and 88,000 Da, respectively), yield value, drug loading, surface morphology and drug release. For in vivo studies, monoarticular arthritis was induced in the left knee joints of rabbits by using ovalbumin and Freund's Complete Adjuvant as antigen and adjuvant. A certain time (4 days) is allowed for the formation of arthritis in the knee joints, then the NS loaded microspheres were injected directly into the articular cavity. At specific time points, gamma scintigrams were obtained to determine the residence time of the microspheres in knee joints, in order to determine the most suitable formulation. This study indicated that PLGA, a synthetic polymer, is more promising than the natural type BSA microspheres for an effective cure of mono-articular arthritis in rabbits. PMID- 11428675 TI - Microencapsulation of gentamicin in biodegradable PLA and/or PLA/PEG copolymer. AB - Biodegradable carriers containing gentamicin for local treatment of bone infection were developed. This paper describes the preparation and in vitro evaluation of these biodegradable implants. Poly-L-lactic acid (PLA) and poly-L lactic acid:polyethylene glycol (PLA/PEG) disk implants containing gentamicin sulphate were obtained by compression of microspheres prepared by a double emulsion process. The mean particle size distribution of the microspheres, based on volume, ranged from 95-270 microm. The gentamicin sulphate loading of the microspheres, after a methylene chloride-water extraction procedure, exceeded 90% of the theoretical value. In vitro dissolution studies on the microspheres and implants with drug loadings 10-40% w/w indicated that the rate of drug release from both PLA and PLA/PEG implants increased, with an increase in drug loading. The release of gentamicin from microspheres was dependent on the properties of PLA and/or PLA/PEG. The PLA/PEG copolymer was more hydrophilic than the PLA homopolymer, and with a smaller pH change in the microenvironment with polymer being degraded. In comparison, the PLA/PEG implant released antibiotic faster and had a larger inhibitory zone based on the Bauer-Kirby experiments used to test the inhibitory activity of antimicrobial devices. Experimental results showed that the biodegradable PLA/PEG gentamicin delivery system had a potential for prophylaxis of post-operative infection. PMID- 11428676 TI - Preparation and in vivo evaluation of parenteral metoclopramide-loaded poly(alkylcyanoacrylate) nanospheres in rats. AB - Metoclopramide-loaded poly(alkylcyanoacrylate) (PACA) nanospheres were prepared by emulsifier-free polymerization in aqueous media at ambient conditions. The optimum polymerization conditions for metoclopramide sorption on PACA nanospheres in the presence of dextran (DEX) or hydroxypropyl-beta-cyclodextran (HPCD) in the polymerization medium were studied and the feasibility of either poly(isobutylcyanoacrylate) (PIBCA) or poly(ethylcyanoacrylate) (PECA) nanospheres as parenteral prolonged release drug delivery system of metaclopramide in rats was also investigated. The optimum time for the addition of metaclopramide after initiating the polymerization was 1 h, which results in 14.8 +/- 0.4 and 9.2 +/- 0.5% of drug loading for PIBCA and PECA, respectively. The HPCD in the polymerization medium of PECA nanospheres improved the drug adsorption compared to DEX at all times, but the difference was only significant (p < 0.05) when metoclopramide was added at 0 and 30 min. Wistar rats were given subcutaneous (s.c.) injections of metoclopramide solution (5 mg/kg) and three different metoclopramide nanospheres suspensions (10 mg/kg) on two phases. The drug solution is rapidly absorbed, distributed, and eliminated. The maximum drug concentration was observed after 30 min of s.c. administration of all the tested nanosphere formulations. PECA-HPCD showed the highest concentration (3.16 +/- 0.66 mg/L) followed by PIBCA-DEX (1.95 +/- 0.37 mg/L) and PECA-DEX (1.68 +/- 0.28 mg/L). The AUCs of PECA-DEX, PECA-HPCD and PIBCA-DEX were 4.8, 1.88 and 2.43 times higher than that of the solution form, respectively. Following PECA-DEX the maximum drug concentration, 1.68 +/- 0.28 mg/L, rapidly decreased to 0.54 +/- 0.05 mg/L. The drug was successfully maintained around this serum drug concentration up to 12 h in rats and the mean drug concentration was reduced to 0.2 +/- 0.02 mg/L, 63% reduction, after 24 h of nanosphere administration. The developed aqueous parenteral prolonged release preparation (PECA-DEX) could be used as a promising intermittent formula for metoclopramide or other drugs when the oral route is not accessible, especially during managing chronic nausea in patients with advanced cancer. PMID- 11428677 TI - A two-step process for controlling the surface smoothness of polyelectrolyte based microcapsules. AB - Biocompatibility is one of the crucial requirements to be fulfilled when designing devices for immunoisolation of transplanted cells. The quality of the capsule surface (smoothness/roughness) influences the nature of cell overgrowth on it by immunocytes, which eventually may lead to the transplant failure. A microcapsule has been developed based on the polyelectrolyte complexation of the polyanions sodium alginate and cellulose sulphate with the polycation poly(methylene-co-guanidine), which was successfully tested in rodent animal models. Recently, the principles for controlling the surface smoothness of these capsules has been identified. This paper reports on a two-step process used for production of stable capsules with improved surface properties. The methodology involves separating the process of drop shape recovery and precursor capsule formation from the process of membrane formation by applying a two-reactor design. The multi-loop reactors are connected in series, and the process separation is given by the different composition of cation solutions flowing in each reactor. This process enables one to prepare the microcapsule immunoisolation device, which can differ in the extent of surface roughness and, thus, is suitable for studying the effect of surface morphology of the immunoisolation device on cell overgrowth. The effect of this process on the capsule permeability has also been evaluated. PMID- 11428678 TI - In vivo perivascular implantation of encapsulated packaging cells for prolonged retroviral gene transfer. AB - Long-term benefits of coronary angioplasty remain limited by the treatment induced renarrowing of arteries, termed restenosis. One of the mechanisms leading to restenosis is the proliferation of smooth muscle cells. Therefore, proliferating cells of the injured arterial wall, which can be selectively transduced by retroviruses, are potential targets for gene therapy strategies. A direct single-dose therapeutic application of retroviral vectors for inhibition of cell proliferation is normally limited by too low transduction efficiencies. Encapsulated retrovirus-producing cells release viral vectors from microcapsules, and may enhance the transduction efficiency by prolonged infection. Primary and immortal murine and porcine cells and murine retrovirus-producing cells were encapsulated in cellulose sulphate. Cell viability was monitored by analysing cell metabolism. Safety, stability, transfer efficiency and extent of restenosis using capsules were determined in a porcine restenosis model for local gene therapy using morphometry, histology, in situ beta-galactosidase assay and PCR. Encapsulation of cells did not impair cell viability. Capsules containing retrovirus-producing cells expressing the beta-galactosidase reporter gene were implanted into periarterial tissue or a pig model of restenosis. Three weeks following implantation, beta-galactosidase activity was detected in the pericapsular tissue with a transduction efficiency of approximately 1 in 500 cells. Adventitial implantation of vector-producing encapsulated cells for gene therapy may, therefore, facilitate successful targeting of proliferating vascular smooth muscle cells, and allow stable integration of therapeutic genes into surrounding cells. The encapsulation of vector-producing cells could represent a novel and feasible way to optimize local retroviral gene therapy. PMID- 11428679 TI - Formulation factors for preparing ocular biodegradable delivery system of 5 fluorouracil microparticles. AB - Microparticles containing 5-fluorouracil (5-FU) were prepared using poly(DL lactide-co-glycolide) with an oil-in-oil emulsion/solvent extraction technique. Particle characteristics including size distribution, 5-FU loading efficiencies, in vitro release and degradation were investigated. The dispersed phase was composed of PLG dissolved in dichloromethane, and the continuous phase was paraffin oil containing lecithin. 5-FU was successfully entrapped in the microparticles with trapping efficiencies up to 76%, loading level 10% w/v, and particle size 3 microm. Release profiles of 5-FU loaded microparticles were determined to follow a first-order-time relationship. An optimized preparation of 5-FU microparticles was achieved and was capable of controlling the release of 5 FU over 21 days with an in vitro delivery rate of 0.4 microg 5-FU/mg particles/day in the study. Preliminary animal studies indicated that the 5-FU loaded microparticles as an ocular delivery system showed no ocular toxicity and no significant inflammatory response in rabbits for 2 months. The 5-FU loaded microparticles approach, with PLG, might be a potential for the application of long-term delivery of hydrophilic drugs in the eye. PMID- 11428680 TI - Preparation of regular sized Ca-alginate microspheres using membrane emulsification method. AB - Monodisperse Ca-alginate microspheres were prepared using the membrane emulsification method. Three ionic types of drugs (anionic, cationic and non ionic) were incorporated into the microspheres, and the effects of sodium alginate concentration and the pressure applied during the dispersing process on the properties of the microspheres were examined. Monodisperse microspheres were obtained when the concentration of alginate solution was 2 wt% and the pressure applied was 0.4 x 10(5) Pa. The mean size of microspheres was approximately 4 microm. Lidocaine x HCl (cationic), sodium salicylate (anionic) and 4 acetamidophenol (non-ionic) were selected as ionic model drugs and included in the alginate microspheres. Lidocaine x HCl (cationic drug) release was more retarded than that of the anionic drug, because of the electrostatic attraction between the negative charge of the ionized carboxyl group in the alginate chain and the positive charge of the cationic drug. In acidic release medium, a slow release was observed due to the low swelling characteristic and the increased viscosity of alginate, regardless of ionic type of drug. PMID- 11428681 TI - Encapsulation of aluminium hydroxide fillers with poly-methyl-methacrylate. AB - A process was developed for the microencapsulation of inorganic filler particles with poly-methyl-methacrylate, to increase the interaction between the hydrophilic filler particles and a polymer matrix. The filler utilised was aluminium hydroxide with an average diameter of 1.9 microm and a specific surface area of 5 m2/g. The process comprised a surface modification, in which a monolayer of isopropoxy titanium isostearate was chemically bound to the surface to render it hydrophobic and to ensure a chemical bond between the filler and the organic phase. Then, an encapsulation reaction was carried out by means of an emulsion-like polymerization process at monomer starved conditions. The modified particles were stabilized in water with sodium-dodecyl-sulphate. A redox system consisting of cumene-hydroperoxide in combination with sodium-formaldehyde sulphoxylate and iron(II) salt was applied for the initiation of the polymerization. Besides surface polymer, free polymer particles were also formed. The parameters which varied were the filler concentration, the concentration of the initiator components and the surfactant concentration. At optimum conditions, approximately 50% of the added monomer polymerized at the modified filler surface, thus forming encapsulated filler particles. SEM together with TGA analysis indicated that a smooth polymer layer had been formed on the filler surface. At high filler loading, however, coagulation occurred. PMID- 11428682 TI - Literature alerts. PMID- 11428683 TI - Characteristics of track cycling. AB - Track cycling events range from a 200 m flying sprint (lasting 10 to 11 seconds) to the 50 km points race (lasting approximately 1 hour). Unlike road cycling competitions where most racing is undertaken at submaximal power outputs, the shorter track events require the cyclist to tax maximally both the aerobic and anaerobic (oxygen independent) metabolic pathways. Elite track cyclists possess key physical and physiological attributes which are matched to the specific requirements of their events: these cyclists must have the appropriate genetic predisposition which is then maximised through effective training interventions. With advances in technology it is now possible to accurately measure both power supply and demand variables under competitive conditions. This information provides better resolution of factors that are important for training programme design and skill development. PMID- 11428684 TI - Physiological characteristics of nationally competitive female road cyclists and demands of competition. AB - There are few published data describing female cyclists and the studies available are difficult to interpret because of the classification of athletes. In this review, cyclists are referred to as either internationally competitive (International Cycling Union world rankings provided when available) or nationally competitive. Based on the limited data available it appears that the age, height, body mass (BM) and body composition of women cyclists who have been selected to the US and Australian National Road Cycling Teams from 1980 to 2000 are fairly similar. Female cyclists who have become internationally competitive are generally between 21 to 28 years of age, 162 to 174 cm, 55.4 to 58.8 kg and 38 to 51 mm (sum of 7 skinfolds) corresponding to 7 to 12% body fat. The lower BM and percentage body fat are traits unique to the most competitive women. Internationally competitive women cyclists also possess a slightly superior ability to produce a high absolute power output for a fixed time period and a noticeably greater ability to produce power output relative to BM. In Women's World Cup races, successful women (top 20 places) spend more time >7.5 W/kg (11 +/- 2 vs 7 +/- 2%, p < 0.01) and less time <0.75 W/kg (24 +/- 4 vs 29 +/- 3%, p = 0.05) compared with non-top 20 placed riders. Additionally, cyclists in the top 20 produced higher average power (3.6 +/- 0.4 vs 3.1 +/- 0.1 W/kg, p = 0.01). Unlike professional men's road cycling, the physiological characteristics of internationally competitive female road cyclists and the demands of women's cycling competition are poorly understood. PMID- 11428685 TI - Physiological and performance characteristics of male professional road cyclists. AB - Male professional road cycling competitions last between 1 hour (e.g. the time trial in the World Championships) and 100 hours (e.g. the Tour de France). Although the final overall standings of a race are individual, it is undoubtedly a team sport. Professional road cyclists present with variable anthropometric values, but display impressive aerobic capacities [maximal power output 370 to 570 W, maximal oxygen uptake 4.4 to 6.4 L/min and power output at the onset of blood lactate accumulation (OBLA) 300 to 500 W]. Because of the variable anthropometric characteristics, 'specialists' have evolved within teams whose job is to perform in different terrain and racing conditions. In this respect, power outputs relative to mass exponents of 0.32 and 1 seem to be the best predictors of level ground and uphill cycling ability, respectively. However, time trial specialists have been shown to meet requirements to be top competitors in all terrain (level and uphill) and cycling conditions (individually and in a group). Based on competition heart rate measurements, time trials are raced under steady state conditions, the shorter time trials being raced at average intensities close to OBLA (approximately 400 to 420 W), with the longer ones close to the individual lactate threshold (LT, approximately 370 to 390 W). Mass-start stages, on the other hand, are raced at low mean intensities (approximately 210 W for the flat stages, approximately 270 W for the high mountain stages), but are characterised by their intermittent nature, with cyclists spending on average 30 to 100 minutes at, and above LT, and 5 to 20 minutes at, and above OBLA. PMID- 11428686 TI - Tests of cycling performance. AB - Performance tests are an integral component of assessment for competitive cyclists in practical and research settings. Cycle ergometry is the basis of most of these tests. Most cycle ergometers are stationary devices that measure power while a cyclist pedals against sliding friction (e.g. Monark), electromagnetic braking (e.g. Lode), or air resistance (e.g. Kingcycle). Mobile ergometers (e.g. SRM cranks) allow measurement of power through the drive train of the cyclist's own bike in real or simulated competitions on the road, in a velodrome or in the laboratory. The manufacturers' calibration of all ergometers is questionable; dynamic recalibration with a special rig is therefore desirable for comparison of cyclists tested on different ergometers. For monitoring changes in performance of a cyclist, an ergometer should introduce negligible random error (variation) in its measurements; in this respect, SRM cranks appear to be the best ergometer, but more comparison studies of ergometers are needed. Random error in the cyclist's performance should also be minimised by choice of an appropriate type of test. Tests based on physiological measures (e.g. maximum oxygen uptake, anaerobic threshold) and tests requiring self-selection of pace (e.g. constant duration and constant-distance tests) usually produce random error of at least approximately 2 to 3% in the measure of power output. Random error as low as approximately 1% is possible for measures of power in 'all-out' sprints, incremental tests, constant-power tests to exhaustion and probably also time trials in an indoor velodrome. Measures with such low error might be suitable for tracking the small changes in competitive performance that matter to elite cyclists. PMID- 11428687 TI - Modelling human locomotion: applications to cycling. AB - Mathematical models of performance in locomotor sports are reducible to functions of the sort y = f(x) where y is some performance variable, such as time, distance or speed, and x is a combination of predictor variables which may include expressions for power (or energy) supply and/or demand. The most valid and useful models are first-principles models that equate expressions for power supply and power demand. Power demand in cycling is the sum of the power required to overcome air resistance and rolling resistance, the power required to change the kinetic energy of the system, and the power required to ride up or down a grade. Power supply is drawn from aerobic and anaerobic sources, and modellers must consider not only the rate but also the kinetics and pattern of power supply. The relative contributions of air resistance to total demand, and of aerobic energy to total supply, increase curvilinearly with performance time, while the importance of other factors decreases. Factors such as crosswinds, aerodynamic accessories and drafting can modify the power demand in cycling, while body configuration/orientation and altitude will affect both power demand and power supply, often in opposite directions. Mathematical models have been used to solve specific problems in cycling, such as the chance of success of a breakaway, the optimal altitude for performance, creating a 'level playing field' to compare performances for selection purposes, and to quantify, in the common currency of minutes and seconds, the effects on performance of changes in physiological, environmental and equipment variables. The development of crank dynamometers and portable gas-analysis systems, combined with a modelling approach, will in the future provide valuable information on the effect of changes in equipment, configuration and environment on both supply and demand-side variables. PMID- 11428688 TI - Adaptations to training in endurance cyclists: implications for performance. AB - Our present scientific knowledge of the effects of specific training interventions undertaken by professional cyclists on selected adaptive responses in skeletal muscle and their consequences for improving endurance performance is limited: sport scientists have found it difficult to persuade elite cyclists to experiment with their training regimens and access to muscle and blood samples from these athletes is sparse. Owing to the lack of scientific study we present a theoretical model of some of the major training-induced adaptations in skeletal muscle that are likely to determine performance capacity in elite cyclists. The model includes, but is not limited to, skeletal muscle morphology, acid-base status and fuel supply. A working premise is that the training-induced changes in skeletal muscle resulting from the high-volume, high-intensity training undertaken by elite cyclists is at least partially responsible for the observed improvements in performance. Using experimental data we provide evidence to support the model. PMID- 11428689 TI - Nutritional practices of male and female endurance cyclists. AB - The nutritional requirements of the training and competition programmes of elite endurance cyclists are challenging. Notwithstanding the limitations of dietary survey techniques, studies of high-level male road cyclists provide important information about nutrient intake and food practices during training and major stage races. Typically, male cyclists undertaking intensive training programmes report a high energy intake (> or = 250 kJ/kg/day) and carbohydrate (CHO) intakes of 8 to 11 g/kg/day. Intakes of protein and micronutrients are likely to meet Recommended Dietary Intake levels, because of high energy intakes. Data on female cyclists are scarce. Stage racing poses an increased requirement for energy and CHO, with daily energy expenditure often exceeding 25 MJ. This must be achieved in the face of practical constraints on the time available for eating, and the suppression of appetite after exhausting exercise. However, studies show that male cyclists riding for professional teams appear to meet these challenges, with the assistance of their medical/scientific support crews. Current dietary practices during cycle tours appear to favour greater reliance on pre-stage intake and post-stage recovery meals to achieve nutritional goals. Recent reports suggest that current riding tactics interfere with previous practices of consuming substantial amounts of fluid and CHO while cycling. Further study is needed to confirm these practices, and to investigate whether these or other dietary strategies produce optimal cycling performance. Other issues that should receive attention include dietary practices of female cyclists, beliefs and practices regarding bodyweight control among cyclists, and the use of supplements and sports foods. PMID- 11428691 TI - Improving cycling performance: how should we spend our time and money. AB - Cycling performance is dependent on physiological factors which influence mechanical power production and mechanical and environmental factors that affect power demand. The purpose of this review was to summarize these factors and to rank them in order of importance. We used a model by Martin et al. to express all performance changes as changes in 40 km time trial performance. We modelled the performance of riders with different ability ranging from novice to elite cyclists. Training is a first and most obvious way to improve power production and was predicted to have the potential to improve 40 km time trial performance by 1 to 10% (1 to 7 minutes). The model also predicts that altitude training per se can cause a further improvement of 23 to 34 seconds. Carbohydrate-electrolyte drinks may decrease 40 km time by 32 to 42 seconds. Relatively low doses of caffeine may improve 40 km time trial performance by 55 to 84 seconds. Another way of improving time trial performance is by reducing the power demand of riding at a certain velocity. Riding with hands on the brake hoods would improve aerodynamics and increase performance time by approximately 5 to 7 minutes and riding with hands on the handlebar drops would increase performance time by 2 to 3 minutes compared with a baseline position (elbows on time trail handle bars). Conversely, riding with a carefully optimised position could decrease performance time by 2 to 2.5 minutes. An aerodynamic frame saved the modelled riders 1:17 to 1:44 min:sec. Furthermore, compared with a conventional wheel set, an aerodynamic wheel set may improve time trial performance time by 60 to 82 seconds. From the analysis in this article it becomes clear that novice cyclists can benefit more from the suggested alterations in position, equipment, nutrition and training compared with elite cyclists. Training seems to be the most important factor, but sometimes large improvements can be made by relatively small changes in body position. More expensive options of performance improvement include altitude training and modifications of equipment (light and aerodynamic bicycle and wheels). Depending on the availability of time and financial resources cyclists have to make decisions about how to achieve their performance improvements. The data presented here may provide a guideline to help make such decisions. PMID- 11428692 TI - Measuring adaptive behavior in inpatient neuropsychiatry: the Behavioural Assessment Scale. AB - The Behavioural Assessment Scale (BAS) was administered to a sample of 95 inpatients with neuropsychiatric conditions. The total scores in the sample ranged from 26 to 145 (maximum possible score is 163), without any evidence of the "floor effect" encountered with other tests used with this population. Investigation into the psychometric structure of the BAS revealed three factors interpreted as Daily Living Skills, Communication/Social Skills, and Problem Behavior. The high intercorrelation between the two dominant factors was interpreted as general sensitivity of the BAS to global decline in functioning associated with severity of illness. These results confirm a previous psychometric investigation carried out on an elderly psychiatric sample. A recommendation is made for interpreting subscales based on these factor domains when specific abilities are of interest and using a Global Functioning subscale as a measure of overall adaptive behavior for both adult and geriatric inpatient neuropsychiatry patients. PMID- 11428690 TI - The effect of altitude on cycling performance: a challenge to traditional concepts. AB - Acute exposure to moderate altitude is likely to enhance cycling performance on flat terrain because the benefit of reduced aerodynamic drag outweighs the decrease in maximum aerobic power [maximal oxygen uptake (VO2max)]. In contrast, when the course is mountainous, cycling performance will be reduced at moderate altitude. Living and training at altitude, or living in an hypoxic environment (approximately 2500 m) but training near sea level, are popular practices among elite cyclists seeking enhanced performance at sea level. In an attempt to confirm or refute the efficacy of these practices, we reviewed studies conducted on highly-trained athletes and, where possible, on elite cyclists. To ensure relevance of the information to the conditions likely to be encountered by cyclists, we concentrated our literature survey on studies that have used 2- to 4 week exposures to moderate altitude (1500 to 3000 m). With acclimatisation there is strong evidence of decreased production or increased clearance of lactate in the muscle, moderate evidence of enhanced muscle buffering capacity (beta m) and tenuous evidence of improved mechanical efficiency (ME) of cycling. Our analysis of the relevant literature indicates that, in contrast to the existing paradigm, adaptation to natural or simulated moderate altitude does not stimulate red cell production sufficiently to increase red cell volume (RCV) and haemoglobin mass (Hb(mass)). Hypoxia does increase serum erthyropoietin levels but the next step in the erythropoietic cascade is not clearly established; there is only weak evidence of an increase in young red blood cells (reticulocytes). Moreover, the collective evidence from studies of highly-trained athletes indicates that adaptation to hypoxia is unlikely to enhance sea level VO2max. Such enhancement would be expected if RCV and Hb(mass) were elevated. The accumulated results of 5 different research groups that have used controlled study designs indicate that continuous living and training at moderate altitude does not improve sea level performance of high level athletes. However, recent studies from 3 independent laboratories have consistently shown small improvements after living in hypoxia and training near sea level. While other research groups have attributed the improved performance to increased RCV and VO2max, we cite evidence that changes at the muscle level (beta m and ME) could be the fundamental mechanism. While living at altitude but training near sea level may be optimal for enhancing the performance of competitive cyclists, much further research is required to confirm its benefit. If this benefit does exist, it probably varies between individuals and averages little more than 1%. PMID- 11428693 TI - A new MMPI-derived indicator of liability to develop schizophrenia: evidence from the New York High-Risk Project. AB - A large body of research indicates that the liability to develop schizophrenia is largely genetically mediated, although phenotypic expression requires environmental triggers/insults and/or epigenetic and/or stochastic factors. In an effort to identify the precise environmental factors that precipitate a predisposition to schizophrenia, researchers have implemented a high-risk model the prospective study of offspring born to schizophrenic parents. As it is difficult to ascertain exactly which of the "high-risk" participants will actually develop the disorder, we examined the validity of an experimental MMPI scale, Schizophrenia Proneness (SzP), and the Moldin-Gottesman psychometric index to identify such individuals. Results suggest that the SzP scale can be an effective predictor of schizophrenia-related psychoses. A revised psychometric index is offered for further study. PMID- 11428694 TI - Revised guidelines for Urist's Mutuality of Autonomy Scale (MOA). AB - Research findings reported in the literature with less than 80% interrater agreement for the Rorschach Mutuality of Autonomy (MOA) scale may be problematic due to possible error. In preparation for a new Rorschach study using the MOA scale, we found it necessary to revise and clarify MOA scale scoring guidelines because they were confusing and ambiguous. A group of 19 naive graduate students improved their interrater agreement scores from 67% using guidelines from the literature, to 75% on the first revision, and these 19 raters plus 13 new raters produced an 80% agreement on a second revision. This revision, with only minor changes based on raters' feedback, is included in an appendix. PMID- 11428695 TI - Increasing the reliability of the fluid/crystallized difference score from the Kaufman Adolescent and Adult Intelligence Test with reliable component analysis. AB - The unreliability of difference scores is a well documented phenomenon in the social sciences and has led researchers and practitioners to interpret differences cautiously, if at all. In the case of the Kaufman Adult and Adolescent Intelligence Test (KAIT), the unreliability of the difference between the Fluid IQ and the Crystallized IQ is due to the high correlation between the two scales. The consequences of the lack of precision with which differences are identified are wide confidence intervals and unpowerful significance tests (i.e., large differences are required to be declared statistically significant). Reliable component analysis (RCA) was performed on the subtests of the KAIT in order to address these problems. RCA is a new data reduction technique that results in uncorrelated component scores with maximum proportions of reliable variance. Results indicate that the scores defined by RCA have discriminant and convergent validity (with respect to the equally weighted scores) and that differences between the scores, derived from a single testing session, were more reliable than differences derived from equal weighting for each age group (11-14 years, 15-34 years, 35-85+ years). This reliability advantage results in narrower confidence intervals around difference scores and smaller differences required for statistical significance. PMID- 11428696 TI - The development and preliminary validation of the Tower of Hanoi-revised. AB - The objective of this study was to redesign the Tower of Hanoi (TOH) in order to increase the reliability and explore the validity of the task. In order to achieve this goal, a two-phase process was undertaken. In Phase 1, the TOH item pool was enlarged from 12 items to 60 items, and the task was administered to a sample of college students (N = 81). The 22 most homogeneous items for the final task were identified based on the item-total correlations. In Phase 2, the rebuilt TOH (TOH-R) was administered to a second sample (N = 50). The internal consistency reliability (Cronbach alpha) was .77 for this new sample. In this phase, the TOH-R was administered concurrently with the Tower of London-Revised (TOL-R), and the convergent validity of the TOH-R against the TOL-R was .53. PMID- 11428697 TI - Psychometric properties and factor structure of the Worry Domains Questionnaire. AB - The Worry Domains Questionnaire (WDQ) is a content-based measure of nonpathological worry. The current study investigates an adapted WDQ for clinical assessment. The WDQ was completed by 570 participants (286 clinical, 284 nonclinical). Internal consistency of the WDQ domains was found to be adequate. Clinical participants obtained significantly higher WDQ scores than control participants. WDQ scores were moderately correlated with several clinical self report measures. A seven-factor structure and a higher order worry factor model were separately subjected to structural equation modeling in the clinical sample. The factor groupings originally outlined by the scale's authors (with minor revisions) provided the best description of the clinical data set. Evidence of differences in the worry factor structures of clinical and control samples did emerge. This study provides initial support of the psychometric suitability of the adapted WDQ for use in clinical populations. PMID- 11428698 TI - Factor index score patterns in the WAIS-III standardization sample. AB - Representative profile subtypes, based on factor index scores, were examined in the standardization sample of the Wechsler Adult Intelligence Scale-Third Edition (WAIS-III). Two-stage cluster analysis identified five reliable subtypes. Three clusters were differentiated primarily by level of performance across all factor index scores (ranging from below average, to average, to above average). The other two clusters were characterized by distinct patterns of performance, with relative efficacy on the Processing Speed index being the most prominent distinction (ranging from a relative weakness to a relative strength). The clusters did not differ significantly in age, but ethnicity and income covaried directly with level of WAIS-III performance, whereas gender affected differential patterns of performance. Furthermore, education and memory were associated primarily with level and, to a relatively minor degree, pattern of WAIS-III performance. Implications for clinical practice are discussed. PMID- 11428699 TI - Comparison of the Fake Bad Scale and other MMPI-2 validity scales with personal injury litigants. AB - Five MMPI-2 validity scales were evaluated with 120 personal injury litigation patients (LP) and 208 clinical patients (CP) along with 43 normal participants (NP). The validity measures included the Fake Bad Scale (FBS), Infrequency scale (F), Back Infrequency scale (Fb), Infrequency-Psychopathology scale (F[p]), and the Dissimulation scale-2 (Ds2). Results showed that only the FBS significantly differentiated the LP and CP, whereas the LP and CP scored significantly higher than the NP on FBS, F, Fb, and Ds2. The content of the FBS, with several items from the Hypochondriasis (Hs) and Hysteria (Hy) Scales, appears to enhance the FBS' ability to detect the somatic overreporting often observed with personal injury claimants. The authors suggest that the FBS may be a useful index of symptom magnification when employed within a comprehensive assessment of malingering in personal injury plaintiffs. PMID- 11428700 TI - MMPI-2 short form: psychometric characteristics in a neuropsychological setting. AB - A 180-item short form of the MMPI-2 (MMPI-2-180) was recently developed by Dahlstrom and Archer and has been proposed for clinical use under special circumstances. This study investigated the psychometric characteristics of the MMPI-2-180 in order to delineate its strengths, limitations, and appropriate scope of clinical application. Using a neuropsychological referral sample (N = 205), we examined accuracy of the short-form as it pertains to the following (a) the prediction of basic scale scores and profile code types, (b) the identification of high-point scales, and (c) the classification of scores as pathological (T > or = 65) or normal range. The results indicate that the MMPI-2 180 provides an unreliable basis for predicting clinical code types, identifying the high-point scale, or predicting the scores on most of the basic scales. In contrast, scores on the MMPI-2-180 are accurate predictors of whether the full scale scores fall within the pathological range (T > or = 65). These findings suggest that (a) standard interpretive procedures involving profile configuration should not be used, in most cases, with the 180-item short-form results, (b) properly interpreted, this shortened version provides potentially useful information regarding the probable presence of various problem areas, and (c) this information is very limited when contrasted with that obtained using the complete or abbreviated (i.e., 370 item) version of the MMPI-2. PMID- 11428701 TI - The detection of malingered posttraumatic stress disorder with MMPI-2 fake bad indices. AB - This investigation explored the effect of posttraumatic stress disorder (PTSD) simulation on Minnesota Multiphasic Personality Inventory-2 (MMPI-2) responses, to detect malingered from genuine PTSD. Sixty-four adult PTSD outpatients at a child sexual abuse (CSA) survivor treatment program were compared with 85 adult college students instructed and trained to malinger PTSD. MMPI-2 overreporting indices examined were F, F-Fb, F-K, F(p), Ds2, O-S, OT, and FBS. A stepwise discriminant analysis identified F(p), F-K, and O-S as the best malingering predictors. A predictive discriminant analysis yielded good hit rates for the model, with impressive cross-validation results. Cutoff scores were assessed for the model's predictors. Clinical implications for detecting malingered PTSD using the MMPI-2 are discussed. PMID- 11428702 TI - Transcriptional influence of two poly purine-pyrimidine tracts located in the HSD11B2 (11beta-hydroxysteroid dehydrogenase type 2) gene. AB - Alternating purine-pyrimidine (APP) sequences which might assume left handed DNA helical structures (Z-DNA) could influence the expression of genes in which they were located. There are two such repeat elements in the HSD11B2 (11beta hydroxysteroid dehydrogenase) gene. First, a CA repeat is located in the first intron. Deletion of this element in a minigene construct leads to a significant decrease (40%) in the expression of the HSD11B2 message in human cortical collecting duct cells. The second APP tract is located approximately 0.9 kb from the last exon or 4.8 kb from the intronic CA repeat element. Deleting this APP tract in a minigene decreases gene expression by 30%. The CA repeat along with its flanking sequences increases luciferase reporter gene expression if placed 5' of the HSD11B2 promoter but not when placed downstream of the reporter gene. Similarly the second APP tract increases luciferase reporter gene expression when placed 5' of the HSD11B2 promoter in an antisense orientation but not in a sense orientation. These results suggests that these dinucleotide repeats influence expression of the HSD11B2 gene in a manner dependent on position and orientation. PMID- 11428703 TI - Impaired testicular function in rats with diet-induced hypercholesterolemia and/or streptozotocin-induced diabetes mellitus. AB - Hypercholesterolemia and diabetes mellitus are known to be accompanied by reproductive dysfunction. In this study, we investigated the effects of hypercholesterolemia, hyperglycemia, and these conditions combined, on testosterone (T) and testicular luteinizing hormone/human chorionic gonadotropin (LH/hCG) binding. Sprague-Dawley rats (8 weeks old) were divided into four groups: Group 1 was the control, group 2 was fed standard chow containing 2% cholesterol (C-diet), group 3 was administered streptozotocin (STZ, 65 mg/kg, i.p.), group 4 was treated with both the C-diet and STZ. After 4 weeks, rats were sacrificed. Serum glucose was significantly higher in the STZ group (304% that of controls) and the C-diet plus STZ group (345%), but there was no difference between the C-diet group (89%) and the control group. Serum cholesterol was significantly higher in the C-diet group (206% that of controls), the STZ group (452%) and the C-diet plus STZ group (2042%). Serum T, testicular T, and LH/hCG binding were significantly lower in the C-diet group (49%, 52%, and 81% that of controls, respectively), the STZ group (15%, 32%, and 72%) and the C-diet plus STZ group (8%, 21%, and 57%). These results suggest that hypercholesterolemia is an independent risk factor for testicular dysfunction and that the reduction of serum and testicular T levels is due at least in part to a reduction in testicular LH/hCG binding in rats with hypercholesterolemia, hyperglycemia, and these conditions combined. It is further suggested that the reduction in LH/hCG binding is mainly related to a rise in serum cholesterol levels. PMID- 11428704 TI - Prolonged cerebellin administration inhibits the growth, but enhances steroidogenic capacity of rat adrenal cortex. AB - Cerebellin is a 16-amino acid peptide, that has been previously found to acutely stimulate steroid secretion from rat adrenal cortex in vivo and in vitro. We have investigated the effects of a prolonged cerebellin treatment (daily injections of 15 nmoles/kg for 6 consecutive days) on the growth and secretion of rat adrenal cortex. Cerebellin lowered adrenal weight, and morphometry showed that this was due to the decrease in the volume of each adrenocortical zone exclusively ensuing from the reduction in the number of its parenchymal cells. Cerebellin did not alter plasma concentration of ACTH, but it raised the levels of circulating aldosterone and corticosterone. The conclusion is drawn that cerebellin chronic administration evokes a marked hypoplastic atrophy of rat adrenocortical cells, that is coupled with an enhanced ACTH-independent steroidogenic capacity of the remaining parenchymal cells. PMID- 11428705 TI - Impact of neonatal hypothyroidism on Leydig cell number, plasma, and testicular interstitial fluid sex steroids concentration. AB - We have previously demonstrated that neonatal and transient neonatal hypothyroidism modulates Leydig, Sertoli, and germ cell numbers, sex steroids and androgen binding protein concentration. The present study was undertaken to study the effect of neonatal onset hypothyroidism on Leydig and peritubular myoid cell numbers, plasma and testicular interstitial fluid (TIF) sex steroid concentration at different age groups of Wistar rats. Hypothyroidism was induced by giving 0.05% methimazole (MMI) to lactating mothers or directly to the male pups from day 1 postpartum through days 10, 15, 30, 40 and 60 postpartum. To confirm hypothyroidism, plasma thyroid hormones and TSH were assayed. Plasma and TIF testosterone, progesterone, dihydrotestosterone (DHT) and estradiol were assayed by radioimmunoassay. Leydig cell number in hypothyroid rats were less than the age-matched controls. The diameter of Leydig cells in hypothyroid rats was smaller than the controls but 10 days old hypothyroids alone had larger than control rats. A significant decrease of peritubular myoid cell number was observed in 30, 40 and 60 days hypothyroid rats and increased in 10 and 15 days hypothyroidism. Hypothyroid rats had elevated level of plasma LH and decreased GH (except day 10 postpartum). Plasma PRL level was increased in 10 and 15 days hypothyroid rats and an opposite effect was observed in 40 and 60 days hypothyroidism. Plasma testosterone, DHT and estradiol were decreased in all hypothyroid rats. However, plasma progesterone level in hypothyroid rats was significantly higher at days 10, 30, and 40 postpartum and an opposite effect was seen in 15 and 60 days experimental groups. TIF testosterone and progesterone titre showed a consistent decrease in hypothyroid rats irrespective of the duration. In hypothyroid rats, TIF DHT levels decreased significantly in days 10, 40 and 60 postpartum. However, it increased in days 15 and 30 postpartum. Except at day 10 postpartum, the level of TIF estradiol in hypothyroid rats was significantly less than their age matched controls. Our data clearly indicate that neonatal onset hypothyroidism adversely affect Leydig cell proliferation along with impaired steroidogenesis. PMID- 11428706 TI - Prostate-thyroid axis: prostatic TRH is one of the stimulators of thyroid hormone. AB - Ventral prostatectomy decreased serum thyroid hormones and histology of the thyroid gland indicate that hypothyroid condition. Co-culture of thyroid gland and ventral prostate stimulates thyroid hormone secretion. In the present study we report prostatic thyrotropin releasing hormone (TRH) is the stimulating factor of thyroid hormone secretion. Mature rat (90 days old) ventral prostate, anterior pituitary and thyroid glands were co-cultured in vitro with or without TRH antibody to assess the direct influence ofprostatic TRH on thyroid hormone secretion. Total thyroxine (T4) and triiodothyronine (T3) were increased significantly in the culture media of ventral prostate, anterior pituitary and thyroid gland when compared with thyroid gland plus anterior pituitary culture media. However, media T4 and T3 concentration decreased significantly in thyroid gland alone; also in thyroid gland plus ventral prostate, thyroid gland plus anterior pituitary and thyroid gland plus anterior pituitary plus ventral prostate were co-cultured with TRH antibody (Ab) in a dose dependent manner. The results suggest that ventral prostatic TRH is one ofthe stimulating factors of thyroid hormone secretion under these in vitro conditions. PMID- 11428707 TI - Androgen-induced cell growth and c-myc expression in human non-transformed epithelial prostatic cells in primary culture. AB - We assessed androgen-induced cell growth and c-myc expression in human non transformed epithelial prostatic (HNTEP) cells in primary culture. Prostatic tissue was obtained from 48 retropubic prostatectomy patients (age: 61-77years) with benign prostatic hyperplasia (malignant tumors excluded). HNTEP cells were treated with testosterone or DHT, alone or in association with hydroxyflutamide. DHT action on c-myc mRNA was examined using Northern blots and RT-PCR. RT-PCR also was used to verify if HNTEP cells expressed the androgen receptor gene. Cell proliferation was assessed on days 3 and 6. Testosterone (2 x 10(-11) M) and DHT (10(-13)M) caused a significant increase (P < 0.05) in cell proliferation on both days. Addition of hydroxyflutamide (10(-6) M) to DHT abolished cell proliferation. HNTEP cells expressed androgen receptor (AR) gene and the treatment with DHT increased AR mRNA levels. C-myc expression was maximal at 30 min and 1 h with DHT (10(-13) M). C-myc seems to play a key role in the control of hormone responsiveness and cell proliferation in epithelial prostatic cells. The detection of androgen receptor gene expression and the increase in this expression with the addition of androgen shows that the HTNEP cells maintain functional characteristics and hormone dependence, and that they are a fruitful in vitro model for studying steroid hormone action mechanisms. PMID- 11428708 TI - Helicobacter pylori infection in Chinese subjects with type 2 diabetes. AB - The relationship between diabetes and Helicobacter pylori (HP) infection is controversial. In this study, we examined the possible relationship between HP infection and type 2 diabetes in Chinese subjects. Sixty-three Chinese type 2 diabetic patients (mean age +/- SD: 49.9 +/- 12.0 years; range: 17-76 years) were recruited irrespective of the duration of diabetes or type of therapy. Twenty nine (46%) of them had upper gastrointestinal symptoms and the other 34 (54%) did not. Another 55 age- and sex-matched non-diabetic subjects (mean age +/- SD: 45.6 +/- 15.6 years, p=0.098; range 18-79 years) with dyspepsia indicated for upper endoscopy were recruited as a comparison group. Upper endoscopy was performed with antral mucosal biopsy specimens taken for rapid urease test (CLO test). HP infection was considered to be present if the rapid urease test was positive. The rates of HP infection of the diabetic and non-diabetic individuals were 50.8% and 56.4% respectively (p: NS). The rate of HP infection was similar between the 2 groups of diabetic patients with or without gastrointestinal symptoms (42.9% vs. 56.3%, p: NS). Using logistic regression analysis (forward stepwise) with age, sex, glycaemic control, duration of diabetes and upper gastrointestinal symptoms as independent variables to predict the risk of HP infection in diabetic patients, none of the parameters enter into the model. In conclusion, the rate of HP infection in Hong Kong Chinese subjects with type 2 diabetes is around 50%, which is similar to control subjects. No association was found between HP infection, glycaemic status, and duration of diabetes and upper gastrointestinal symptoms in these diabetic subjects. PMID- 11428709 TI - Prevalence and comparisons of five different diagnostic methods for Helicobacter pylori in diabetic patients. AB - Helicobacter pylori is now regarded as a major gastroduodenal pathogen that is etiologically linked with duodenal and gastric disease. It has been suggested recently as an important factor for nongastroenterologic conditions such as coronary heart disease and diabetes mellitus. In this study, we planned to investigate the prevalence of H. pylori in diabetic patients and to evaluate five different diagnostic tests. Group I consisted of 67 patients with type II diabetes mellitus and seventy-three aged-matched health people served as control in group II. Group I was divided in two subgroups with good (Group IA) and poor (Group IB) glycemic control. H. pylori was diagnosed by five different tests: 1) biopsy, 2) culture, 3) gram staining, 4) imprint cytology and 5) brushing cytology. The usefulness of each test for each group was statistically compared. There was a higher prevalence for H. pylori in diabetic patients. This study showed that two positive out of five tests was most reliable for predicting the H. pylori in diabetic and nondiabetic patients. In conclusion, the prevalence of H. pylori is high in diabetic patients. Peristaltic activity, and impaired nonspecific immunity must be evaluated as risk factors in diabetics. We recommend that the 'gold standard' should be regarded as two positive out of these five different tests. PMID- 11428710 TI - Stress from exercise in the below sea level environment causes an increase in serum testosterone levels in trained athletes. AB - A comparative study (n = 20) of serum levels of leutinizing hormone (LH) and testosterone (T) between male trained athletes (high-school students, 16-18 years old) living in North and South Shouna in the Jordan Valley (JV), 320-360 meters below sea level, and those living in Ramtha and Irbid, 550-650 meters above sea level, was conducted in November, 1999. Serum levels of LH and T were also measured in these athletes following a 20 Km noncompetitive run. The air temperatures on the day of the experiment was comparable (25 degrees C in North and South Shouna versus 23 degrees C in Ramtha and Irbid). Before exercise, serum levels of LH and T in athletes of the below sea level environment (North and South Shouna) were similar to those levels in athletes of the above sea level environment (Ramtha and Irbid). Exercise caused a significant increase in serum levels of both LH and T only in athletes of North and South Shouna. It seems likely that the higher secretion of LH contributes, at least in part, for the higher serum levels of T following exercise. Taken together, these data suggest that exercise has an effect on LH and T secretion that is similar to that of fasting. And finally, the below sea level environmental factors, such as the high barometric pressure, as well as the genetic background of the athletes affect the pituitary and adrenal cortex as well as testicular tissue, resulting in the secretion of more LH and T. PMID- 11428711 TI - Effect of G-CSF on ethanol-induced hemorrhagic gastritis model in diabetes mellitus-induced rats. AB - Diabetes mellitus can affect every organ system, including large and small vessels, eyes, nerves, kidneys and gastrointestinal system. Acid peptic disease is an inflammatory condition involving the upper gastrointestinal tract. The elevated serum glucose levels of diabetics affect traditional host defenses such as neutrophil counts and functions. We aimed to investigate changes of gastric mucosa and the role of impaired neutrophil functions in a diabetes-induced experimental model and whether G-CSF, which modulates neutrophil counts and function, has protective effects against gastric mucosal injury in diabetic rats. Fifty rats were divided into three groups. Diabetes mellitus was induced by a single dose of streptozotocin in 40 of 50 rats. Controls had a sham injection. The gastric mucosal lesions were produced by intragastric administration of 1 ml of 95% ethanol in all three groups. Granulocyte colony-stimulating factor (G-CSF) was subcutaneously injected to twenty of diabetes-induced rats. Stomach histology and tissue malondialdehyde and glutathione levels were determined. White blood cell count, neutrophil counts and functions were determined. Peripheral blood cell counts, neutrophil phagocytosis index were decreased but neutrophil adhesivity index was not different in diabetes-induced groups. G-CSF administration improved netrophil counts and function. Macroscopic and microscopic gastric mucosal injury were significantly greater in control and only diabetes group compared with G-CSF pretreated group (p < 0.05). The tissue malondialdehyde and glutathione levels were significantly decreased in G-CSF administrated diabetic group compared to untreated diabetics (p < 0.001). Finally, G-CSF has been shown to cause neutrophilia and improve neutrophil phagocytosis in diabetic. G-CSF may be cytoprotective for gastric mucosa in diabetes mellitus-induced rats. PMID- 11428712 TI - Early changes in parameters of bone and mineral metabolism during therapy for hyper- and hypothyroidism. AB - The effects of thyroid hormones on various organs and metabolic systems have been the focus of intensive research. In this study we investigated the mechanisms of the changes in some parameters of bone and mineral metabolism before and during treatment of hyper- and hypothyroidism. Our study groups were as follows; 1) Untreated hyperthyroid patients (n= 38), 2) Hyperthyroid patients treated for three months (n=21), 3) Untreated hypothyroid patients (n=27), 4) Hypothyroid patients treated for three months (n= 20), and 5) Euthyroid control subjects (age, weight, sex and menopausal status matched) (n = 47). As expected, the mean serum calcium (Ca), phosphorus (P), alkaline phosphatase (ALP), and urinary Ca/creatinine and deoxypyridinoline (D-Pyr)/creatinine levels were higher in group-1 than in the control group. Serum PTH level was lower in group-1 than in group-5. However, after treatment for three months (group-2) we found that the serum and urinary levels of these parameters (except ALP) were not different than in the control group. Group-3 and group-4 did not show any differences in these parameters compared with group-5. Covariance analysis showed that urinary D-Pyr excretion had a positive, independent relationship to the serum free T3 level and age (P < 0.001 and P = 0.02, respectively). These results suggest that both bone formation and resorption markers increase in hyperthyroid patients, and with the treatment, particularly, in the period of first three months the bone resorption markers decrease rapidly. If the treatment is maintained the decrease slows, becoming more gradual. However, bone formation markers like ALP remain high in hyperthyroid patients during the treatment. In the light of this data, it is possible to conclude that osteoblastic activity lasts longer in hyperthyroidism. On the other hand, we demonstrated that these bone formation and resorption markers do not seem to be different in hypothyroid patients, even during the treatment, compared to the euthyroid controls. PMID- 11428713 TI - Effect of chronic morphine administration on glucose tolerance and insulin binding to isolated rat adipocytes. AB - Morphine addiction was induced in six male Wistar rats. Improved glucose tolerance (peak value less by 28%, p<0.01) was observed in chronically morphinized rats as compared to the control rats, injected with saline. An increase in the maximal specific binding of 125I-labeled insulin to unit membrane area of adipocytes was observed in the experimental group (p < 0.01). The changes in insulin receptor number could be responsible for the improved glucose tolerance observed during morphine addiction. PMID- 11428714 TI - Progesterone induced modulations of serum hormonal profiles in adult male and female rats. AB - The impact of progesterone on serum hormonal profiles in the presence and absence of gonads was studied in adult male and female albino rats. Progesterone was administered intramuscularly for 30 days at a dose of 1 mg/100g body weight/day. Serum testosterone, estradiol and prolactin titres decreased in male and female rats with intact gonads given progesterone. While the levels of both luteinizing hormone (LH) and follicle stimulating hormone (FSH) decreased in male rats with intact gonads, only FSH decreased in female rats. The inhibitory effect of progesterone on serum estradiol, LH, FSH and prolactin persisted even after gonadectomy in male rats. This persistent inhibitory effect of progesterone was also seen on serum testosterone, FSH and prolactin levels of female rats. Ovariectomy modified progesterone action on LH, as is evident from the decreased levels of LH observed only in ovariectomized rats given progesterone. While progesterone had no effect on serum T3 and T4 in male rats, gonadectomy altered the levels of T3 and T4 in male and female rats. Progesterone increased the levels of T3 and decreased the levels of T4 in ovariectomized rats. Growth hormone (GH) and thyroid stimulating hormone (TSH) levels seem to be resistant to changes in progesterone titre, irrespective of the sex and gonadal status. The present data suggest the existence of a sex specific effect of progesterone on gonadotrophins. The data on T3, T4 and TSH reveals that progesterone has no effect on the pituitary thyroid axis in the presence of gonads. PMID- 11428715 TI - cAMP-induced apoptosis in granulosa cells is associated with up-regulation of P53 and bax and down-regulation of clusterin. AB - Evidence indicates that cAMP induces apoptosis in granulosa cells of rat and human ovary. The mechanism by which cAMP induces apoptosis is not known. This study was carried out to evaluate changes in expression of cell death promoters, P53 and bax, and cell death repressor, bcl-2, in cAMP-treated granulosa cells. Treatment of granulosa cells with forskolin (FSK), or 8-bromo-cAMP induced apoptosis as evidenced by internucleosomal DNA fragmentation and chromatin condensation as revealed by gel electrophoresis and fluorescent DAPI staining, respectively. The apoptotic effect of cAMP was accompanied by an increase in the expression of P53 and bax proteins as evaluated by Western blot and immunocytochemistry. No change in bcl-2 protein level was observed in cAMP treated granulosa cells as compared to control. These data suggest that cAMP may activate apoptosis in granulosa cells by shifting the ratio of the death promoter to death repressor genes via alteration of P53 and bax expression. cAMP was also shown to inhibit gene expression of clusterin, an apoptosis-associated protein, suggesting a role for this protein in cAMP-induced apoptosis in granulosa cells. The data of the present study provide a basis for future studies to elucidate the molecular mechanism of follicular atresia and regulation of apoptotic cell death in ovarian follicles. PMID- 11428716 TI - Evidence of different actions of testosterone, estradiol, FSH, and LH on the growth axis. AB - To study the possible role of sexual hormones, Testosterone (T), Estradiol (E2), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) on the growth axis, we examined the correlations between the sex, growth hormone (GH), insulin like growth factor-1 (IGF-1), insulin-like growth factor binding protein 3 (IGFBP3), FSH, LH, T and E2, in growth retarded children ranging in age from 7 to 13 yr. All hormones were measured by Radioimmunoassay (RIA) in a pool of aliquots of samples obtained every 20 min over 12 h (overnight) in each child. Multivariate statistical analysis was performed. We have found: a) Thai only FSH concentrations were significantly higher in girls than in boys; b) A positive correlation between T-IGF-1; T-IGFBP3; FSH-LH; FSH-IGF-1; FSH-IGFBP3, LH-IGF-1, LH-IGFBP3 the sex-FSH; and the sex-IGFBP3 c) A high positive correlation between plasmatic E2 and IGF-1/IGFBP3 ratio (an index of free, active IGF-1). We concluded that the sex, FSH, LH, T and E2 influence the growth axis. The sex through IGFBP3; LH, FSH, and T through IGF1 and IGFBP3; E2 through the IGF 1/IGFBP3 ratio, an index of active IGF-1. PMID- 11428717 TI - The influence of short-term fasting on serum leptin levels, and selected hormonal and metabolic parameters in morbidly obese and lean females. AB - The aim of our study was to compare the changes of serum leptin levels after 24-h fasting in morbidly obese and lean females and to search for hormonal and metabolic factors responsible for the changes in serum leptin levels. Fourteen morbidly obese and twelve lean females were included in the study. The blood for leptin, insulin, cortisol, blood glucose (BG), beta-OH-butyrate (beta-OH), dehydroepiandrosterone (DHEA) and DHEA-sulphate (DHEA-S) measurements was withdrawn before and after a 24-h fast. Basal body mass index (BMI), serum leptin, insulin and beta-OH levels were significantly higher in the obese compared to the lean group. The 24-h fasting decreased significantly BMI, serum leptin (by 20% in obese vs. 62% in lean subjects), insulin (by 23.3% in obese vs. 23.1% in lean subjects) and increased beta-OH (by 36% in obese vs. 1300% in lean subjects). Basal serum leptin levels correlated positively with BMI in both groups and with insulin levels in the obese group. The multiple regression analysis using delta leptin as dependent and the basal values of the rest of studied parameters as independent variables revealed that in lean subjects serum cortisol together with DHEA-S and BMI accounted for 71% of variations of the change of serum leptin levels (delta leptin = 0.31- 0.0101 cortisol + 0.0012 DHEA S + 0.37 BMI). In obese subjects the 43.9% of variations of the change of serum leptin levels was explained by BMI together with age and DHEA-S levels (delta leptin = 36.09 + 0.35 BMI - 0.717 age- 0.008 DHEA-S). The drop of serum leptin levels after 24-h starvation is significantly blunted in obese compared to lean subjects. The reason for the difference is probably the insulin resistance possibly further modified by different DHEA-S levels. PMID- 11428718 TI - Urinary free cortisol excretion pattern in morbid obese women. AB - The urinary excretion of free cortisol in a group of 10 control and 20 morbidly obese women was measured in all bladder voidings during 24 h. The data from obese women were measured under Hospital basal controlled conditions and after 3 days of very low calorie diet (VLCD, 1.9 MJ/d). The hourly cortisol excretion pattern was determined for each woman, and means of each group were computed in order to obtain a 24 h excretion pattern. In controls, the highest excretion rate was in the morning (8-9 h) and the lowest at 21-22 h. Inbasal conditions, the obese showed a similar but flatter pattern; the highest peak was also in the morning (9 10 h), but the lowest rate was between 21 and 24 h. The VLCD diet flattened the pattern even more, in away that no clear peak was observed from the early morning until the afternoon; however, the nadir coincided with that found in basal conditions. These patterns resulted in significant differences between VLCD, basal diet and control. The amount of free cortisol excreted was 93.0 +/- 6.9 nmol/ day in controls, 70.1 +/- 4.7 nmol/day in obese under basal conditions and 62.6 +/- 3.0 nmol/day when subjected to VLCD. The results presented are consistent with a lower overall cortisol secretion in the morbid obese women, which also show a narrower margin of variation in cortisol secretion than non obese controls. The data also show the significant influence of dietary energy on the pattern of cortisol excretion in obese women. PMID- 11428719 TI - Crystalline dihydrotestosterone implants in the lateral septum of male rats. A positive effect on LH and FSH. AB - Previous investigations in our laboratory have shown that testosterone implanted into the lateral septum in male rats increases LH and FSH secretion. However, it was unclear whether the effect of testosterone was direct via androgen receptor, or indirect via the estrogen receptor after conversion by aromatization to estradiol. To answer this question, we implanted either testosterone or the non aromatizable androgen 5 alpha-dihydrotestosterone (DHT), into the lateral septum of adult male rats and measured plasma levels of LH and FSH by radioimmunoassay 2 days after implantation. Both testosterone and DHT significantly increased the plasma LH and FSH concentrations. Mean concentration of LH in control animals was 0.21 +/- 0.06 ng/ml, a figure that increased to 0.7 +/- 0.12 and 0.55 +/- 0.1 ng/ml after DHT or testosterone implantation respectively. Mean concentration of FSH in control animals was 1.5 +/- 0.3 ng/ml; this figure increased to 3 +/- 0.3 and 2.9 +/- 0.3 ng/ml after DHT or testosterone implantation. Neither plasma DHT (64.0 +/- 5.6 vs. 52 +/- 5 ng/100ml) nor plasma testosterone levels (4.1 +/- 0.38 vs. 3.3 +/- 0.18 ng/ml) were significantly affected by the implants. We conclude that androgens independently of conversion to estrogen acting in the lateral septum facilitates the release of LH and FSH. PMID- 11428720 TI - Unrecognized persistence of beta-hydroxybutyrate in diabetic ketoacidosis. AB - We quantitated serial serum beta-hydroxybutyrate (beta-OHB) levels using the Ketosite method in 9 children with IDDM who were treated for diabetic ketoacidosis (DKA) and compared them to urinary ketone measurements by dipstick. Persistent elevations of serum beta-OHB were seen in six patients when the urine became clear of ketones; five of these patients had a recurrence of ketonuria. We conclude that many patients recovering from ketoacidosis have continuing elevations of beta-OHB after the urine is free of ketones and this unrecognized abnormality is the likely cause of recurrence of the ketonuria. We recommend that fluid therapy be continued beyond clearance of ketonuria and suggest using the Ketosite method to document restoration of normal serum beta-OHB levels in patients recovering from DKA. PMID- 11428721 TI - Lack of mutations of type 1 11beta-hydroxysteroid dehydrogenase gene in patients with abdominal obesity. AB - There is increasing evidence that in human obesity, particularly the abdominal phenotype, the activity of the hypothalamic-pituitary-adrenal (HPA) axis is disregulated. At least two distinct alterations have been reported: one is characterized by several neuroendocrine abnormalities and hyperresponsiveness of the HPA axis to different neuropeptides, the other is characterized by elevated cortisol traffic and probably by supranormal cortisol production. The 11beta hydroxysteroid dehydrogenase (11beta-HSD) enzymes interconvert cortisol and cortisone in human. Two different isoforms have been identified. A possible modification of the activity of the enzyme 11beta-HSD1 in subjects with abdominal obesity has been described in the literature. We decided to test the hypothesis that mutated isoforms of type 11beta-HSD1 protein could be responsible for alterations of cortisol metabolism in patients with abdominal obesity. A mutational screening of the whole coding sequence and exon-flanking regions of the 11B-HSD1 gene has been performed in 8 patients. The main results of our study are the exclusion of a common association of 11beta-HSD1 mutations to obesity and the identification of two novel allelic variants for the gene 11beta-HSD1 in the Italian population, not previously described in any database. PMID- 11428722 TI - Hepatocyte nuclear factor-1alpha inhibits insulin promoter factor 1-dependent transactivation of the human insulin gene. AB - To investigate the regulational interaction of hepatocyte nuclear factor-1alpha (HNF-1alpha) and insulin promoter factor 1 (IPF1) on insulin gene expression, either or both of the expression vectors carrying each transcription factor were transiently transfected into HeLa cells, RINm5F cells and MIN6 cells together with the luciferase reporter construct driven by a human preproinsulin gene promoter (-1998 to +237) designated as, pINS-1998/luc. IPF1-transfection into HeLa cells strongly stimulated the luciferase activity to 725 fold that of the basal level. In contrast, HNF-1alpha-transfection resulted in only a 6.7 fold increase. In co-transfection experiments, increasing the amount of HNF-1alpha resulted in an 84.5% and 74.4% decrease in IPF1-stimulated luciferase activity in HeLa and RINm5F cells, respectively. Deletion constructs designated as pINS 248/luc, pINS-213/luc and pINS-185/luc were transfected into RINm5F cells to determine the role of the A3 element and its 5' flanking sequence in the inhibitory effect of HNF-1alpha. The results showed that the inhibiting effects of HNF-1alpha with pINS-213/luc and pINS-185/luc were significantly smaller than those with both pINS-1998/luc and pINS-248/luc. Transfection into MN6 cells with pINS-1998/luc in the absence of IPF1 resulted in constitutional transactivation of the insulin gene, and this transactivation was abolished by the co transfection with HNF-1alpha. The present data indicate that IPF1 rather than HNF 1alpha predominantly transactivates the insulin gene, and that HNF-1alpha inhibits IPF1-dependent insulin gene transactivation mediated through the 5' flanking sequence of the A3 element. It is suggested that HNF-1alpha may be involved in insulin gene expression as a negative regulator. PMID- 11428723 TI - Sertoli cells inhibited apoptosis of pachytene spermatocytes and round spermatids. AB - Apoptosis in the testis represents an important physiological mechanism that regulates the number of germ cells in the seminiferous epithelium. This apoptosis is believed to be regulated by many factors, including growth factors and cytokines, which appear to suppress apoptosis of the germ cells. In this study, we examined the roles of Sertoli cells on the regulation of pachytene spermatocyte (PS) and round spermatid (RSd) apoptosis with either a co-culture trans-well system or a direct contact system. Apoptosis was detected by low molecular weight DNA fragmentation assay, in situ end labeling, and an LDH assay. In addition, the level of Bcl-2, Bax, and ICE mRNAs in PS and RSd by Northern blot analysis. When PS and RSd were cultured with Sertoli cells in either a trans well system or direct contact system, the extent of apoptotic DNA fragmentation and LDH level were both significantly lower than those control values. TUNEL staining also revealed the inhibition of apoptosis of PS and RSd when they were cultured with Sertoli cells compared with controls (p <0.05). Moreover, the extent of apoptotic DNA fragmentation and LDH level were significantly lower in the direct contact system than in the trans-well system. TUNEL staining also demonstrated a more decrease in apoptosis of PS and RSd in the direct contact system compared with the trans-well system (p < 0.05). PS and RSd cultured with Sertoli cells exhibited an increase in Bcl-2 mRNA, whereas those cultured with serum-free medium did not show any change. The levels of Bax and ICE mRNAs decreased in PS and RSd cultured with Sertoli cells in comparison with control values. These results suggest that Sertoli cells can prevent apoptosis of germ cells, and that the effect of Sertoli cells on germ cells is mediated by cell to cell interaction or, remote effects of inhibitory factors on apoptosis. PMID- 11428724 TI - Plasma ferritin and type 2 diabetes mellitus: a critical review. AB - Multiple factors appear to be involved in the pathogenesis of type 2 (non insulin dependent) diabetes mellitus (DM). One of these factors may be iron overload. This critical review summarizes the major studies on the link between type 2 DM, insulin resistance, glycemic control, diabetic complications and hyperferritinemia. Although some studies suggested that plasma ferritin concentration is positively correlated with insulin resistance and with the risk of acquiring type 2 DM, substantial iron overload is not a typical feature of DM. There is no correlation between plasma ferritin level and glycemic control or diabetic microangiopathic complications. PMID- 11428725 TI - The DD genotype of the ACE gene polymorphism is associated with diabetic nephropathy in the type-1 diabetics. AB - Genetic factors are involved in the development of diabetic nephropathy in type-1 diabetes. We are examining the association of the angiotensin-converting enzyme (ACE), insertion/deletion (I/D) polymorphism with the presence of diabetic nephropathy in type-1 diabetic patients. 52 type-1 diabetic patients with diabetic nephropathy (30 with either microalbuminuria or macroalbuminuria and 22 with end stage renal disease on dialysis) were compared with 10 type-1 diabetic patients with normoalbuminuria and duration of disease longer than 15 years and 27 non-diabetic healthy subjects. We found that the D-allele frequency was higher in patients with nephropathy than in the healthy and normoalbuminuric controls. There was an association in the DD polymorphism of the ACE gene with patients with diabetic nephropathy and not with the control subjects. We conclude that the DD genotype of ACE gene polymorphism is associated with diabetic nephropathy in patients with type-1 diabetes mellitus. PMID- 11428726 TI - Progress toward poliomyelitis eradication--West and Central Africa, 1999-2000. AB - In 1988, the World Health Assembly of the World Health Organization (WHO) resolved to eradicate poliomyelitis by 2000. Reported polio cases have decreased on all continents. In 2000, poliovirus was isolated from 24 countries, 13 in the African Region of WHO (AFR). This report summarizes the routine polio vaccination coverage, surveillance for acute flaccid paralysis (AFP) during 1999 and 2000, and the synchronization of national immunization days (NIDs) against polio during 2000 and early 2001 in 16 countries in west and central Africa. PMID- 11428727 TI - Exposure to patients with meningococcal disease on aircrafts--United States, 1999 2001. AB - Neisseria meningitidis is a leading cause of bacterial meningitis and sepsis in children and young adults in the United States and is spread through direct contact with respiratory secretions. Persons in close contact with patients who have meningococcal disease are at increased risk for contracting the disease. Commercial aircraft are suitable environments for the spread of airborne pathogens, including N. meningitidis. A case of air-travel-associated meningococcal disease is defined as a patient who meets the case definition of meningococcal disease within 14 days of travel on a flight of at least 8 hours duration. Because of concerns about disease transmission aboard aircraft, CDC has developed recommendations to ensure a standard approach to management of airline contacts. This report presents a case of air-travel-associated meningococcal disease and presents guidelines for the management of persons potentially exposed to meningococcus during airtravel. PMID- 11428728 TI - University outbreak of calicivirus infection mistakenly attributed to Shiga toxin producing Escherichia coli O157:H7--Virginia, 2000. AB - On February 21-22, 2000, the Virginia Department of Health (VDH) was notified by a university student health center of two suspected cases of Escherichia coli O157:H7. At a local hospital laboratory, stool specimens from the two ill students tested positive for Shiga toxin-producing E. coli (STEC) using a commercially available enzyme immunoassay (EIA) kit. Further investigation revealed that the outbreak of gastrointestinal illness was caused by a Norwalk like virus (NLV), a member of the calicivirus family. This report summarizes the outbreak investigation and laboratory findings used to identify the causative agent, and highlights the need for follow-up cultures on all specimens testing positive for STEC by EIA and for submission of isolates to state laboratories so that public health agencies can respond appropriately in identifying common source outbreaks. PMID- 11428729 TI - Kernicterus in full-term infants--United States, 1994-1998. AB - Kernicterus is a preventable life-long neurologic syndrome caused by severe and untreated hyperbilirubinemia during the neonatal period. High levels of bilirubin are toxic to the developing newborn. In full-term infants, hyperbilirubinemia symptoms include severe jaundice, lethargy, and poorfeeding. Features of kernicterus may include choreoathetoid cerebral palsy, mental retardation, sensorineural hearing loss, and gaze paresis. Kernicterus is not a reportable condition in the United States, and its prevalence is unknown; however, a pilot registry at a Pennsylvania hospital documented 90 cases in 21 states from 1984 to June 2001 (L. Johnson, Pennsylvania Hospital, Philadelphia, personal communication, 2001). This report summarizes case histories of four full-term, healthy infants who developed kernicterus and underscores that to prevent kernicterus, newborns must be screened and promptly treated for hyperbilirubinemia. PMID- 11428730 TI - Self-injectable epinephrine: no size fits all! PMID- 11428731 TI - Corticosteroid dosing and asthma control. PMID- 11428732 TI - Churg-Strauss syndrome. AB - OBJECTIVES: This article discusses the clinical manifestations and treatment protocols of Churg-Strauss syndrome (CSS). A review of the definition, pathophysiology, and prognosis of CSS is included, as well as more recent evidence of the presumed association between antileukotriene antagonists and CSS. DATA SOURCES: Relevant articles in the medical literature derived from searching the MEDLINE database (1966 to present) with key terms Churg-Strauss syndrome, allergic granulomatosis, and allergic granulomatous angiitis. Sources included review articles, meta-analyses, randomized control trials, case reports, case series, and seminal articles, the majority of which had been published within the past decade. STUDY SELECTION: Studies that described the clinical manifestations, pathophysiology, etiology, treatment, or prognosis of CSS. RESULTS: CSS is a systemic vasculitic disorder with multiorgan involvement and diverse presentations. CONCLUSIONS: Recognition of the multiorgan manifestations of CSS is crucial to clinical management. Whether a causal relationship exists between antileukotriene antagonists and onset of CSS remains unclear. PMID- 11428733 TI - Epinephrine dispensing for the out-of-hospital treatment of anaphylaxis in infants and children: a population-based study. AB - BACKGROUND: Epinephrine is life-saving in the treatment of anaphylaxis. A limited number of fixed-dose epinephrine formulations are available for out-of-hospital treatment of this disorder. OBJECTIVE: To examine dispensing patterns for epinephrine formulations over 4 consecutive years in a population of 279,638 infants, children, and adolescents (from birth up to but not including the 17th birthday). METHODS: We used the Drug Programs Information Network, an administrative claims database for prescriptions dispensed in ambulatory care settings, developed from real-time computer links with retail pharmacies in the province of Manitoba, Canada. We studied the specific epinephrine formulation dispensed and the precise age of the infant or child at the time it was dispensed. RESULTS: Epinephrine formulations were dispensed for 1.2% of the pediatric population (3,340 children). Boys comprised 59.5% of the recipients. Of all epinephrine formulations, 38.6% were dispensed as EpiPen Jr (0.15 mg), and 57.4% were dispensed as EpiPen (0.3 mg). EpiPen Jr was dispensed for patients ranging in age from 2 months to 16 years, 10 months, inclusive. EpiPen was dispensed for patients ranging in age from 1 year, 8 months to 16 years, 11 months, inclusive. During the 4 years studied, a subgroup of children transitioned from EpiPen Jr to EpiPen auto-injectors at a mean age of 6 years, 6 months +/- 2 years, 8 months (range 1 year, 10 months to 16 years, 11 months). CONCLUSIONS: Both EpiPen Jr and EpiPen auto-injectors were dispensed over almost the entire age range of the pediatric population. Physicians should consider a child's age more carefully when prescribing these auto-injectors. Additional concentrations of epinephrine are needed in these fixed-dose formulations. PMID- 11428734 TI - Once-daily budesonide via Turbuhaler improves symptoms in adults with persistent asthma. AB - BACKGROUND: Previous studies have demonstrated the efficacy and safety of twice daily budesonide Turbuhaler (Pulmicort Turbuhaler, AstraZeneca, Wilmington, DE) for the treatment of mild to severe asthma. OBJECTIVE: To compare the efficacy and safety of budesonide Turbuhaler administered once daily each morning with placebo in inhaled corticosteroid-naive adults with persistent asthma. METHODS: In this randomized, double-blind, placebo-controlled, multicenter study, 177 adults (aged 18 to 70 years) received placebo or once-daily budesonide Turbuhaler (400 microg) for 12 weeks. Efficacy variables included mean changes from baseline in forced expiratory volume in 1 second (FEV1) and AM/PM peak expiratory flow rate (PEFR), and nighttime/daytime asthma symptom scores, patient discontinuations, use of breakthrough medication (albuterol), forced vital capacity (FVC), forced expiratory flow between 25% and 75% of FVC (FEF25%-75%), and quality of life assessments. Safety was evaluated based on adverse events, physical examinations, vital signs, and laboratory tests. RESULTS: Demographic and baseline characteristics were comparable between study groups. The mean percentages of predicted FEV1 at baseline were 71.9 +/- 9.8 in patients receiving budesonide Turbuhaler and 70.6 +/- 11.0 in patients receiving placebo. Mean changes from baseline over the 12-week treatment period in FEV1 were significantly (P = 0.007) improved in patients receiving once-daily budesonide Turbuhaler compared with placebo (0.31 L and 0.17 L, respectively). Significant (P < or = 0.037) improvements over placebo also were observed in AM PEFR, nighttime/daytime asthma symptoms, and albuterol use with budesonide Turbuhaler treatment. Adverse events were generally mild or moderate in intensity and similar between study groups. CONCLUSIONS: Budesonide Turbuhaler 400 microg administered once daily in the AM is efficacious and safe for inhaled corticosteroid-naive asthmatic adults. PMID- 11428735 TI - Once-daily budesonide inhalation powder (Pulmicort Turbuhaler) maintains pulmonary function and symptoms of asthmatic children previously receiving inhaled corticosteroids. AB - BACKGROUND: The incidence of pediatric asthma has increased dramatically over the past few decades, with approximately 5% of American children affected by the disease. OBJECTIVES: To compare the efficacy and safety of once-daily budesonide Turbuhaler with placebo in asthmatic children previously treated with orally inhaled corticosteroids. METHODS: This randomized, double-blind, placebo controlled, multicenter (17 centers) study included 274 male and female children (aged 6 to 17 years) with a history of asthma for at least the previous 6 months. Patients received placebo or budesonide Turbuhaler (200 microg or 400 microg) once daily for 12 weeks. Efficacy variables included mean changes from baseline in forced expiratory volume in 1 second (FEV1), AM and PM peak expiratory flow rates (PEFRs), nighttime and daytime asthma symptom severity scores, patient discontinuations, use of beta2-agonists as breakthrough medication, forced vital capacity (FVC), and midexpiratory flow rate between 25% and 75% of FVC (FEF25% 75%). Safety was evaluated by adverse events, physical examinations, vital signs, and laboratory tests. RESULTS: Baseline characteristics were comparable among treatment groups. Percentage of predicted FEV1 at baseline was 76.6 +/- 6.9 for placebo, 77.5 +/- 7.1, and 77.0 +/- 7.8 for the budesonide Turbuhaler 200 microg and 400 microg groups, respectively. Significantly (P < or = 0.024) more placebo patients (24%) discontinued treatment because of disease deterioration or no improvement than budesonide Turbuhaler 200 microg (11%) or 400 microg patients (10%). Patients receiving budesonide Turbuhaler experienced significant improvements in FEV1 compared with patients receiving placebo (P < or = 0.015). Significant (P < or = 0.041) improvements over placebo also were observed in AM and PM PEFRs, FVC, FEF25%-75%, nighttime and daytime asthma symptoms, and amount of beta2-agonist used in both budesonide Turbuhaler groups. Adverse events were generally mild or moderate in intensity and similar among treatment groups. CONCLUSIONS: Once-daily budesonide Turbuhaler is effective and safe in children with persistent asthma previously maintained on at least twice-daily dosing regimens of inhaled corticosteroids. PMID- 11428736 TI - Olopatadine ophthalmic solution adjunctive to loratadine compared with loratadine alone in patients with active seasonal allergic conjunctivitis symptoms. AB - BACKGROUND: Olopatadine ophthalmic solution 0.1% (Patanol, Alcon Laboratories, Fort Woth, TX) is approved for the treatment of the signs and symptoms of allergic conjunctivitis. Loratadine 10 mg (Claritin, Schering-Plough, Madison, NJ) is a nonsedating oral antihistamine approved for the treatment of the signs and symptoms of allergic rhinitis. OBJECTIVE: To compare the efficacy of olopatadine used adjunctively with loratadine versus loratadine alone in patients with seasonal allergic conjunctivitis. METHODS: This three-center, observer masked, treatment-controlled, randomized, parallel-group study involved patients aged 7 to 74 years with seasonal allergic conjunctivitis. Patients were treated for 7 days with either olopatadine twice daily adjunctive to loratadine once daily or only loratadine once daily. Efficacy variables (ocular itching and redness, physician's impression, patient's impression, patient diary ratings of ocular redness and itching), and safety parameters were evaluated during the screening visit and on days 0, 3, and 7. Patients completed the rhinoconjunctivitis quality of life questionnaire on days 0 and 7. RESULTS: Ninety-four patients received study drug. Patients receiving olopatadine twice daily in addition to loratadine once daily exhibited less ocular itching (P = 0.0436) and rated their ocular condition as more improved compared with those receiving loratadine alone (P < 0.0022). Twenty minutes after initial dosing, olopatadine plus loratadine relieved ocular itching and redness significantly better than loratadine alone (P = 0.001). Both treatment groups showed clinically meaningful improvements in overall quality of life in all but one of the rhinoconjunctivitis quality of life questionnaire domains. Overall, and in most domains, olopatadine plus loratadine also provided significantly better (P < 0.05) quality of life than loratadine alone at day 7. CONCLUSIONS: Compared with loratadine alone, olopatadine adjunctive to loratadine provides greater relief of ocular itching and redness, a better quality of life, and is well tolerated in patients with seasonal allergic conjunctivitis. PMID- 11428737 TI - Safety of inhaled corticosteroid therapy in young children with asthma. AB - BACKGROUND: Physicians have had some reluctance to use inhaled corticosteroids in very young children with asthma because of the possible risks of adverse systemic effects. OBJECTIVE: The purpose of this study was to evaluate the effects of fluticasone propionate on growth and adrenocortical function in young children with asthma. METHODS: We performed an open, prospective study for 24 weeks of 20 children with asthma, 2.5 to 5.0 years of age, who had received fluticasone by a large volume spacer at dosages ranging from 190.50 to 565.40 microg/m2 daily. Growth was evaluated by height standard deviation scores measured by a stadiometer. Adrenocortical function was evaluated twice in each child, before and after the study, by determining fasting serum cortisol concentrations at 8 AM and also at 30 and 60 minutes after adrenocorticotropic hormone stimulation. Posttreatment values of height standard deviation scores and fasting morning serum cortisol concentrations were compared with those of 18 age-matched children, who constituted the control group. RESULTS: The evaluation of mean +/- SEM (and range) of height standard deviation scores revealed a significant decrease from 0.44 +/- 0.27 (-1.46 to 2.22) to 0.28 +/- 0.26 (-1.51 to 2.07; P = 0.01) at week 18 and to 0.25 +/- 0.24 (-1.90 to 2.13; P = 0.04) at the week 24 in fluticasone-treated children. At the end of the treatment, however, height standard deviation scores of these children did not differ significantly (P = 0.35) from those of the control group. Delayed growth with medium-duration treatment was not associated with alterations in serum cortisol measurements, either at baseline or after stimulation. The mean fasting morning serum cortisol concentrations did not differ significantly between the fluticasone-treated patients and the control group. CONCLUSIONS: Some concern prevails about the safety of medium- or long-term treatment with regularly inhaled corticosteroids in young children with asthma. The prepubertal growth may be delayed, but the effect on ultimate height remains uncertain in such children. Growth should be regularly monitored in children who begin inhaled corticosteroid therapy for mild persistent asthma at an age <5 years old. PMID- 11428738 TI - Comparison of montelukast versus budesonide in the treatment of exercise-induced bronchoconstriction. AB - BACKGROUND: Previous studies in which leukotriene-receptor antagonist and corticosteroids were used have suggested a possible role for these anti inflammatory drugs in the prevention of exercise-induced bronchoconstriction, but no direct comparisons have been made. OBJECTIVE: A crossover study was undertaken to compare the ability of both montelukast and budesonide to protect patients from exercise-induced bronchoconstriction. METHODS: A total of 20 patients (median age, 17 years; range, 8 to 36 years), who had clinical exercise-induced bronchoconstriction for 1 year and decreased FEV1 of at least 20% after exercise on two occasions, were enrolled in this study. To compare the therapies in each patient, we administered, consecutively, 10 mg of montelukast once daily at bedtime for 3 days and, later, 400 microg of budesonide twice daily for 15 days, or vice versa, with a 15-day intervening washout period during which no patient received treatment. Exercise challenges were performed at baseline (no therapy) and after each treatment. The percentage of FEV1 declines at 2, 7, and 12 minutes after exercise and the area under the curve (summarizing the extent and modification of FEV1 decreases relative to time) were measured and compared. RESULTS: Both budesonide and montelukast significantly reduced the decrease in FEV1 (area under the curve) after exercise with respect to the baseline condition of no therapy (P = 0.0001). Overall, budesonide offered better protection (area under the curve) than did montelukast (P = 0.01), particularly in the short-term evaluation (2 minutes after exercise; P = 0.003); however, considerable individual variations in the responses to both budesonide and montelukast were observed. The degree of protection against decreases in FEV1 ranged from 0% to almost 100% for both treatments. In 16 of 20 patients, budesonide therapy offered better protection than did montelukast, and in the other 4 patients, montelukast showed better protection than did budesonide. No side effects of either montelukast or budesonide were detected during the study. CONCLUSIONS: Treatment with budesonide or montelukast prevents exercise-induced bronchoconstriction. Because substantial variation in the response may be present among patients, both drugs should be tested in each patient before long-term therapy is chosen. PMID- 11428739 TI - Distribution and cytokine production of CD4 and CD8 T-lymphocyte subsets in patients with acute asthma attacks. AB - BACKGROUND: The activation of T cells and the elevation of Th2-type cytokines have been observed in asthmatic patients, but the relative role of CD4 and CD8 T cell is still unclear. OBJECTIVE: To investigate the role of T cell subset in patients with acute asthma attacks, we analyzed the distribution, activation status, and cytokine production of CD4 and CD8 cells. METHODS: The percentages of the CD4 and CD8 cell in peripheral blood (PB) and bronchoalveolar lavage (BAL) fluid were analyzed by flow cytometry. The cytokines (IL-4, IL-5, and IFN-gamma) and soluble IL-2 receptor (sIL-2R) were measured by ELISA in culture supernatants of CD4 and CD8 cells purified from PB. RESULTS: The CD4/CD8 ratio in PB of asthmatic patients was significantly higher than that of controls, which was significantly reduced after treatment. In contrast, there was a tendency to high percentage of CD8 cells in asthmatic patients as compared with controls in BAL, which resulted in a decreased CD4/CD8 ratio. Comparing the T cell subsets in BAL with paired PB in asthma, the CD4 cells were higher in PB, but CD8 cells were higher in BAL. The IL-4, IL-5, and sIL-2R produced by CD4 cells were significantly higher than those produced by CD8 cells in asthmatic patients. CONCLUSIONS: Our results provide evidence that activated CD4 T cells increase and produce type 2 cytokines in PB, but CD8 T cell are more sequestrated than CD4 T cells in the airway during an acute asthma attack. PMID- 11428740 TI - Duodenal IgE-positive cells and elimination diet responsiveness in children with atopic dermatitis. AB - BACKGROUND: Parameters for identifying eczematous children who could respond to an elimination diet are needed. In children with food allergy, duodenal IgE containing cells are enhanced. OBJECTIVE: To determine the presence of duodenal mucosal IgE-positive cells in atopic dermatitis and to determine whether duodenal IgE-positive cells may identify eczematous children who will benefit from an elimination diet. METHODS: Thirty-one children with severe eczema underwent gastrointestinal endoscopy because of gastrointestinal symptoms and were treated with an elimination diet. A clinical score to skin lesions was given before and after diet. All subjects were skin-prick tested with food antigens and aeroallergens. Serum IgE levels were measured. Duodenal IgE-positive cells were investigated in 18 control subjects and in all eczematous children before diet. RESULTS: The number of duodenal IgE-positive cells in children with atopic dermatitis was significantly increased compared with that of control group (P < 0.001). Nineteen (61%) eczematous children improved on a few food diet. Diet responsive children had significantly higher IgE-positive cells compared with both nondiet-responsive and controls. Positive predictive accuracy of duodenal IgE-positive cells was poor, whereas negative predictive accuracy was high at the cutoff level of 50 IgE-positive cells/10 visual fields. Diagnostic accuracy both of SPT reactions with foods and of food-specific serum IgE antibodies was poor. CONCLUSIONS: An intestinal IgE-mediated reaction occurred in children with severe atopic dermatitis who underwent intestinal endoscopy because of gastrointestinal symptoms. In these eczematous children, the number of IgE-positive cells in the duodenal mucosa might be helpful for excluding a positive response to the elimination diet. PMID- 11428741 TI - Tacrolimus inhibits cytokine production and chemical mediator release following antigen stimulation of passively sensitized human lung tissues. AB - BACKGROUND: The immunosuppressive effects of tacrolimus are mediated by inhibition of cytokine production by inflammatory cells. The role of tacrolimus on cytokine production and release of chemical mediators in asthma is not known at present. OBJECTIVES: We compared the effects of tacrolimus on interleukin (IL) 5 and tumor necrosis factor-alpha (TNF-alpha) production and chemical mediator release from excised human lung tissue with those of steroids. METHODS: Human lung tissue was passively sensitized with serum from atopic patients then preincubated with tacrolimus (10(-6), 10(-7), 10(-8) M) or dexamethasone (10(-6) M) for 2 hours. The lung tissue was then exposed to 1.5 microg/mL of mite antigen and then cultured for 48 hours. Culture supernatants were collected and IL-5 and TNF-alpha levels were measured by ELISA. IL-5 and TNF-alpha messenger ribonucleic acid (mRNA) expression was also investigated by reverse transcriptase-polymerase chain reaction. The level of histamine and leukotriene E4 was also measured in the culture supernatant. In addition, tryptase staining was performed to compare degranulation of mast cells. RESULTS: Antigen stimulation increased histamine and leukotriene release in the supernatant. Tacrolimus significantly and dose dependently inhibited the release of histamine and leukotriene; dexamethasone did not. The results of tryptase staining demonstrated that tacrolimus dose dependently inhibited degranulation of mast cells, whereas dexamethasone did not. Antigen stimulation increased TNF-alpha and IL-5 protein production and mRNA expression. Tacrolimus and dexamethasone significantly inhibited TNF-alpha and IL 5 protein production and mRNA expression. CONCLUSIONS: Our results indicated that tacrolimus is more powerful in inhibition of cytokine production and release of chemical mediators than steroids, and suggested that this immunosuppressor drug might be useful for the treatment of asthma. PMID- 11428742 TI - Anisakis simplex-sensitized patients: should fish be excluded from their diet? AB - BACKGROUND: Anisakis simplex (A.s.) allergy is an emerging disease. The third stage larvae of this nematode are a source of hidden allergens in fish. There are no clear guidelines concerning dietary restrictions for patients with serum specific IgE to this parasite. OBJECTIVE: To follow up the clinical data and immunological parameters of patients sensitized to A.s. during 6 to 23 months. METHODS: The clinical symptoms and serologic status of 17 patients with specific IgE and positive skin prick test results to A.s. were studied prospectively. Six of these had anaphylaxis (ANA) attributed to A.s. and 11 patients experienced concomitant chronic urticaria (CU). All patients were advised not to eat fish for 6 months. RESULTS: Four patients from the ANA group excluded fish, and ANA did not recur. Two other patients with ANA refused to exclude fish; one remained free of symptoms and the other experienced several urticarial episodes. During this 6 month period total IgE levels decreased in all six ANA patients; specific IgE for A.s. decreased in four patients and increased in two. Two patients from the CU group did not exclude fish, and symptoms persisted in these two patients. Clinical improvement was observed in 78% of the patients with CU who excluded fish. Total and specific IgE levels decreased in all the patients with CU. CONCLUSIONS: Because ANA symptoms are very severe, patients should always be advised to exclude fish until specific food allergens are identified. However, in patients with CU and specific IgE to A.s., only the clinical response to fish ingestion will determine the need for strict fish avoidance. PMID- 11428743 TI - The diagnostic value of crude or boiled extracts to identify tolerant versus nontolerant lentil-sensitive children. AB - OBJECTIVE: The aim of this study was to compare two types of lentil extracts for use in skin prick tests for the diagnosis of lentil clinical allergy. METHODS: Thirty-six patients with a history of allergic reactions after the ingestion of lentils were skin tested with two types of lentil extracts at 0.05, 0.5, 5, and 10 mg/mL. Both extracts were extracted at 40 degrees C and afterward, one of them was boiled for 15 minutes. Thirty-three of these patients underwent oral challenges with lentils and three had a convincing recent history of lentil anaphylaxis. RESULTS: Twenty patients had a positive oral challenge; 13 were negative. Skin prick tests performed with the boiled extract at 0.5 and 5 mg/mL were positive in 96% and 100% of patients with positive food challenge, and in 31% and 85% of those with negative food challenge, respectively; positive skin test results were similar in both groups using the crude extract. Mean wheal sizes using the boiled extract at 0.5, 5, and 10 mg/mL were significantly greater in patients with a positive oral challenge than in those with a negative one (4.9, 6.8, and 7.4 mm versus 1.9, 3.5, and 5.1 mm, respectively; P < 0.05) These mean values were not statistically different using the crude extract. CONCLUSIONS: These data suggest that lentil extracts for the diagnosis of lentil hypersensitivity should be heated, since boiled extracts, used at a concentration of 0.5 or 5 mg/mL, best identify clinically sensitive individuals. PMID- 11428744 TI - Differences in sensitization rates to outdoor aeroallergens, especially citrus red mite (Panonychus citri), between urban and rural children. AB - BACKGROUND: A recent investigation has suggested that citrus red mite (Panonychus citri, CRM) is the most important allergen in citrus-cultivating farmers with asthma and allergic rhinitis. OBJECTIVE: A cross-sectional survey was performed to evaluate the prevalence of asthma and chronic rhinitis symptoms and sensitization to common indoor and outdoor aeroallergens, including CRM and Japanese cedar pollen, in rural and urban Korean children. METHODS: A total of 2,055 children (1,055 subjects living in rural areas with citrus farms and 1,000 controls in urban areas without citrus farms) were enrolled. They were evaluated by a questionnaire, and by skin prick tests with 13 common indoor and outdoor aeroallergens, including CRM and Japanese cedar pollen. RESULTS: The prevalence of wheezing and chronic rhinitis symptoms during the last 12 months was 8.3% and 35.7% in the rural children and 10.5% and 22.4% in the control group. The most common sensitizing allergens in order of decreasing frequency were Dermatophagoides pteronyssinus (26.6%), Dermatoplagoides farinae (22.7%), CRM (14.2%), cockroach (11.3%), and Japanese cedar pollen (9.7%) among the rural children, but the sensitization rates to CRM and Japanese cedar pollen were 1.3% and 0.2% among the control children, respectively. The prevalence of wheeze during the last 12 months was not different between rural children with sensitization to CRM or Japanese cedar pollen and those without sensitization (5.4% vs 6.1%; 6.9% vs 5.9%). However, the prevalence of chronic rhinitis during the last 12 months was higher among those with sensitization to CRM or to Japanese cedar pollen than among those without sensitization (40.8% vs 34.4%; 51.5% vs 33.5%). CONCLUSIONS: CRM is a common sensitizing allergen in rural children, and the sensitization rates to outdoor aeroallergens, especially CRM and Japanese cedar pollen, are very different between children from rural and urban areas in Korea. PMID- 11428745 TI - Chronic urticaria and Helicobacter pylori. AB - BACKGROUND: Although the clinical manifestations of chronic urticaria (CU) are similar in most patients, a variety of factors should be taken into consideration. In general, the cause of CU cannot be determined in most patients, and it is considered idiopathic. In the past several years, relationships between some patients with CU and hepatitis C or autoimmune thyroid diseases have been established. Similarly, other factors may also be considered as possible causes to explain certain patients with CU. Previously, some patients with CU have had their disease attributed to Helicobacter pylori (HP), but the relationship was only clinical. OBJECTIVE: None of the patients previously described included an immunological study. Thus, we studied a patient with CU, who showed marked clinical improvement after eradication of HP, to demonstrate an IgE relationship with this skin disease. METHODS: First, blood analytical parameters, roentgenograms, fecal examination for parasites, and skin tests were performed to try to establish an etiology. In addition, endoscopy with gastric biopsy confirmed HP colonization, and eradication treatment was prescribed. To investigate an immunological relationship, other tests performed included the following: HP-specific IgG, histamine release induced by HP, HP-specific IgE, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis with immunoblotting. RESULTS: The blood analytical parameters, roentgenograms, fecal examination for parasites, and skin tests were all negative. In contrast, the tests for HP specific IgG, histamine release induced by HP, and HP-specific IgE were all positive. In addition, the sulfate-polyacrylamide gel electrophoresis with immunoblotting showed specific IgE binding to an extract of HP. CONCLUSIONS: Our results may indicate an immunological IgE relationship between HP colonization and CU in this particular patient. PMID- 11428746 TI - Natural history of adult asthma. PMID- 11428747 TI - The expiration reflex from the vocal folds. AB - The authors present their 30 years' experience with expiration reflex. The reflex can be elicited from vocal folds by mechanical, chemical or electrical stimulation of the superior laryngeal nerve of man and laboratory animals, except mice and rats. It manifests itself by a short, forcible expiratory effort without a preceding inspiration which is indispensable for cough effort. The role of expiration reflex is to prevent penetration of foreign bodies into airways, expelling phlegm and detritus from subglottal area. The initial inspiration before expiration is undesired and could lead to inspiration pneumonia. The reflex is well known to laryngologists as '"laryngeal cough." Its receptors are small in number, localised mainly in medial margin of vocal folds deep in mucosa which can explain their stability in pathological conditions of the larygx. Afferentiation of the reflex is via laryngeal nerve similarly to sneezing and cough. Expiration reflex is not co-ordinated by a single "centre" but rather by a network system in the brain stem. Its motor pattern is supposedly produced by "multifunctional" population of medullar neurones in Botzinger complex and the rostral ventral respiratory group involved also in the genesis of breathing and cough. However, in cats also other neurones may play a vital role in production, shaping and mediation of the motor pattern of respiratory reflex, localised in rostral pons, lateral tegmental field or in the raphe medullar midline. PMID- 11428748 TI - A hybrid on-line ECG segmenting system for long-term monitoring. AB - This paper introduces a new hybrid ECG beat segmenting system, which can be applied in the processing unit of single-channel, long-term ECG monitors for the on-line segmentation of the ECG signal. Numerous ECG segmentation techniques are already existing and applied, however sufficiently robust and reliable methods currently require more than one ECG signal channel and quite complex computations, which are practically not feasible in stand-alone, low-cost monitors. Our new system approach presents a time domain segmentation technique based on a priori physiological and morphological information of the ECG beat. The segmentation is carried out after classifying the ECG beat, using the linear approximation of the filtered ECG signal and considering the pathophysiological properties as well. The proposed algorithms require moderate computational power, allowing the practical realization in battery powered stand-alone long-term cardiac monitors or small-sized cardiac defibrillators. The prototype version of the system was implemented in Matlab. The test and evaluation of the system was carried out with the help of reference signal databases. PMID- 11428749 TI - The effect of moderate weight loss on echocardiographic parameters in obese female patients. AB - Based on the data collected by KNOLL Hungary Ltd. in Hungary in 1999, 37% of the adult population is overweight while 23% is obese. Inappropriate diet containing excess calories and physical inactivity are responsible for these statistical values. In their former studies, the authors investigated the effects of different stages of obesity on the cardiovascular system, and have verified that even moderate obesity elicits pathological geometric and functional changes in the heart. In the present study, effect of a half-year-long life-style modification program on the morphologic and functional characteristics of the heart was investigated in twenty-one obese women. Life-style modification contained a diet with reduced energy uptake (1000-1300 Cal/day) and a regular physical training of minimum 3-4 hours weekly. By the end of the sixth month the weight loss was 5.1 kg (5.9%) on an average. There was a marked reduction in cardiac dimensions measured by echocardiography, with a very slight, non significant decrease in left ventricular internal diameter, and a marked, significant reduction in the left ventricular wall thickness. Decrease of the left ventricular muscle mass exceeded the decrease of body weight. A marked elevation was found in the E/A quotient that reflected a definite improvement in diastolic function. Results indicate that physical training programs have a favourable effect on the echocardiographic parameters, therefore the process is reversible even without a pharmacological intervention. PMID- 11428750 TI - Effect of glycerol-induced acute renal failure and di-2-ethylhexyl phthalate on the enzymes involved in biotransformation of xenobiotixs. AB - The effects of di-(2-ethylhexyl)-phthalate (DEPH) on the levels of cytochrome P 450 and b5 monooxygenases were studied in the rat kidney and liver in acute renal failure induced by glycerol. Intramuscular injection of glycerol (50%,10 ml x kg( 1)) to rats produced proximal tubular damage and acute renal failure. The indicators of renal function, serum urea and creatinine significantly increased (480 and 350 percent, respectively). In control and glycerol-treated animals DEPH had no significant effect on the concentrations of serum urea and creatinine. Twenty-four hours after glycerol injection the total amount of cytochrome P-450 and b5 significantly decreased in renal but increased in liver microsomal fractions. Moreover, 48 and 72 hours after glycerol injection the level of cytochrome P-450 and b5 significantly increased in both organs. A single dose of DEPH (2 ml x kg(-1), i.p.) also elevated the total cytochrome P-450 and b5 in control animals. This enhancing effect of DEPH was additive to that of glycerol in glycerol-induced acute renal failure. These results indicate that DEPH and glycerol evoked pathological changes may affect the metabolism of xenobiotics plus endogenous hormones in the liver and in kidney. PMID- 11428751 TI - Aerobic and anaerobic training effects on the antioxidant enzymes of the blood. AB - The purpose of the present study was to investigate the effects of aerobic and anaerobic training on serum lipid peroxidation levels and on antioxidant enzyme activities. Long distance runners for aerobic training group, and wrestlers for anaerobic training group were chosen. Non-sporting men were used as control group. When the aerobic power was compared; indirect VO2max of long-distance runners were found higher than wrestlers and control group (p<0.001, p<0.001). When lipid peroxidation levels were compared; levels of the thiobarbituric acid reactive substances (TBARS) of long distance runners were found to be lower than those in the control group (p<0.05), but similar to those found in wrestlers. Comparison of antioxidant enzyme activities in erythrocytes show that there were no significant difference among the groups in superoxide dismutase enzyme activities, but glutathione peroxidase (GPx) activity of long distance runners was higher than that measured in wrestlers (p<0.05). These results suggest that aerobic training increased in erythrocytes GPx activity with a subsequent decrease in plasma TBARS levels but anaerobic training had no effect on this process. PMID- 11428752 TI - Coexistence of at least three distinct beta-adrenoceptors in human internal mammary artery. AB - The internal mammary artery (IMA) is currently the preferred conduit for myocardial revascularization. However, perioperative vasospasm and a hypoperfusion state during maximal exercise may limit its use as a bypass graft. The mechanism of spasm has not been clearly defined. Since beta-adrenoceptor activation plays a major role in vasorelaxation, the present study was carried out to investigate the beta-adrenoceptor responsiveness of human IMA smooth muscle. Isoproterenol produced a concentration-dependent relaxation in endothelium-denuded IMA segments, precontracted with phenylephrine (maximal relaxation 46.33+/-5.45%). Atenolol (10(-6)M) and propranolol (2x10(-7)M) inhibited isoproterenol-induced relaxation. While atenolol produced partial inhibition, propranolol caused a complete inhibition in a majority of the segments and a partial inhibition in a minority. BRL 37344, a selective beta 3 adrenoceptor agonist, produced a concentration-dependent relaxation in phenylephrine-precontracted rings of endothelium-denuded IMA (maximal relaxation 40.35+/-4.07%). Cyanopindolol, a beta-adrenoceptor partial agonist, produced a marked relaxation (58.65+/-6.2%) in endothelium-denuded IMA rings, precontracted with phenylephrine. Cyanopindolol-induced relaxation was resistant to blockade by propranolol (2x10(-7)M). Spontaneous contractions of IMA rings were also observed in some cases that were inhibited by isoproterenol and BRL 37344. This observation implies the important role of beta-adrenoceptor activation in prevention of human IMA spasm. The results obtained in present study indicate that human IMA smooth muscle possesses an atypical beta-adrenoceptor together with beta1- and beta2-adrenoceptors. Regarding the relaxation induced in IMA rings by adding BRL 37344, the possible identical entities of IMA atypical beta adrenoceptors and beta 3-adrenoceptors are suggested. PMID- 11428753 TI - Eudaimonia. PMID- 11428754 TI - Roger A. Mann Award . The reliability of angular measurements in hallux valgus deformities. . AB - The purpose of this study was to determine the intraobserver and inter-observer reliability of physicians on a repetitive basis in making angular measurements of hallux valgus deformities. The hallux valgus angle, the 1-2 intermetatarsal angle, and the distal metatarsal articular angle and the assessment of congruency/subluxation of the first MTP joint were evaluated on a repetitive basis. Physicians were provided with a series of black and white photographs of radiographs with a hallux valgus deformity. Three different sets of photographs randomly ordered were sent at a minimum interval of six weeks to the participants. Participating physicians were extremely reliable in the measurement of the 1-2 metatarsal angle. 96.7% of the photographs were repeatedly measured within a range of 5 degrees or less. The angular measurements to determine the hallux valgus angle were slightly less reliable, but 86.2% of photos were repeatedly measured within a range of 5 degrees or less. In the measurement of the distal metatarsal articular angle, 58.9% of photographs were repeatedly measured within a range of 5 degrees or less. There was a wide range within physician evaluators who recognized very few congruent joints (2 of 21) and those who recognized several congruent joints (11 of 21). Most physicians appeared to be internally consistent in the assessment of MTP congruency; however, some photographs were much more difficult to assess than others. This study validates the reliability of the measurement of the hallux valgus and the 1-2 metatarsal angle. The interobserver reliability in the measurement of the distal metatarsal articular angle is questioned. PMID- 11428755 TI - Treatment of type V osteochondral lesions of the talus with ipsilateral knee osteochondral autografts. AB - Treatment of symptomatic large cystic lesions of the talus has had mixed results. A technique of treatment using a cored osteochondral graft taken from the ipsilateral knee is presented. Preliminary results in ten consecutive patients show significant improvement in all patients with an average increase of 27 points in the AOFAS Hindfoot score. PMID- 11428756 TI - Osteochondral defects of the talus treated with fresh osteochondral allograft transplantation. AB - Between 1980 and 1996, 9 patients with osteocartilagenous lesions of the talus were treated surgically using fresh osteochondral allograft transplantation. In 8 cases the reason for surgery was osteochondritis dissecans (4 of these cases had a previous traumatic injury). In 1 case a fresh osteochondral allograft of the talus was required following a traumatic open fracture of the talus sustained in a motor vehicle accident. Of these 9 grafts, 6 grafts remain in situ with a mean survival of 11 years (range 4 to 19). In the three cases requiring fusion the reason for surgery was not related to arthritic deterioration but due to resorption and fragmentation of the graft. PMID- 11428757 TI - Fractures and fracture dislocations of the midfoot: occurrence, causes and long term results. AB - Etiology and outcome of 155 patients with midfoot fractures between 1972 and 1997 were analyzed to create a basis for treatment optimization. Cause of injuries were traffic accidents (72.2%), falls (11.6%), blunt injuries (7.7%) and others (5.8%). Isolated midfoot fractures (I) were found in 55 (35.5%) cases, Lisfranc fracture dislocations (L) in 49 (31.2%), Chopart-Lisfranc fracture dislocations (CL) in 26 (16.8%) and Chopart fracture dislocations (C) in 25 (16%). One hundred and forty eight (95%) of the midfoot fractures were treated operatively; 30 with closed reduction, 115 with open reduction, 3 patients had a primary amputation. Seven (5%) patients were treated non-operatively. Ninety seven (63%) patients had follow-up at an average of 9 (1.3-25, median 8.5) years. The average scores of the entire follow-up group were as follows: AOFAS - sum of all four sections (AOFAS-ET): 296, AOFAS-Midfoot (AOFAS-M): 71, Hannover Scoring System (HSS): 65, and Hannover Questionnaire (Q): 63. Regarding age, gender, cause, time from injury to treatment and method of treatment no score differences were noted (t test: p>0.05). L, C or I showed similar scores and CL significantly lower scores (AOFAS-ET, AOFAS-M, HSS, Q). The highest scores in all groups were achieved in those fractures treated with early open reduction and operative fixation. Midfoot fractures, particularly fracture dislocation injuries, effect the function of the entire foot in the long-term outcome. But even in these complex injuries, an early anatomic (open) reduction and stable (internal) fixation can minimize the percentage of long-term impairment. PMID- 11428758 TI - Results of operative fixation of unstable ankle fractures in geriatric patients. AB - It is widely accepted that operative fixation of unstable ankle fractures yields predictably good outcomes in the general population. The current literature, however reports less acceptable results in the geriatric population age 65 years and older. The current study analyzes the outcome of the surgical treatment of unstable ankle fractures in patients at least 65 years old. Twenty three patient over 65 years old were surgically treated after sustaining 21 (91%) closed and 2 (9%) open grade II unstable ankle fractures. Fractures were classified according to the Danis-Weber and Lauge-Hansen schemes. Fracture type was predominantly Weber B (21/23, 91%), or supination external rotation stage IV (21/23, 91%). Fracture union rate was 100%. There were three significant complications including a lateral wound dehiscence with delayed fibular union in an open fracture dislocation, and two below knee amputations, neither of which was directly related to the fracture treatment. There were three minor complications; one superficial wound infection and two cases of prolonged incision drainage, all of which resolved without further surgical intervention. Complications were associated with open fractures and preexisting systemic disease. These results indicate that open reduction and internal fixation of unstable ankle fractures in geriatric patients is an efficacious treatment regime that with results that are comparable to the general population. PMID- 11428759 TI - Why do ankle arthrodeses fail in patients with rheumatic disease? AB - Solid and painless fusion was achieved in 117/130 patients (90%) with rheumatic diseases after primary ankle arthrodesis at the authors' institution. Operations were performed using internal fixation according to the Adams technique. Critical retrospective analysis of failures in 13 patients (11 nonunions, one postoperative low-grade infection, and one painful arthrodesis) revealed errors in the primary operative technique in 10/13 ankles (77%), resulting typically from the surgeon's attempt to overcompensate a malaligned ankle while ignoring correction of the hindfoot deformity (subtalar complex). The optimum of 0-5 degrees of valgus was found in only 5/13 patients (38%). All four patients with varus alignment presented with malleolar pain. Bone grafting was adequate even in those patients with failure, whereas immobilization time was suboptimal in one patient (eight weeks). Patient satisfaction was lowered in every case of nonunion. Revision arthrodesis of failed primary fusion was successful in 10/13 patients (77%), however three additional stress fractures, two painful ankles without nonunions, and one superficial wound infection were detected. Ankle arthrodesis is a demanding procedure, and the operation should always be performed by an experienced surgeon, taking into account the alignment, ligament, and muscle balance of the rheumatoid ankle and hindfoot. Correction and rebalancing of these factors and the use of bone grafts are of crucial importance when considering the optimal conditions for fusion. Nonunions, infections, and stress fractures occurring after the primary arthrodesis are severe complications, leading eventually to revision operations and problems with osteoporotic bone, fragile soft tissues, and skin. PMID- 11428760 TI - Rubber foreign bodies in puncture wounds of the foot in patients wearing rubber soled shoes. AB - We report 8 cases of puncture wound of the foot associated with rubber foreign bodies in patients who were wearing rubber-soled shoes. The difficulty in making the correct diagnosis and the complications arising from these injuries are reviewed. The morbidity associated with these seemingly innocuous puncture wounds can be serious. Infective complications resolved only with removal of all imbedded rubber foreign bodies. A history of wearing rubber-soled shoes during the injury and a high index of suspicion may prevent complications. PMID- 11428761 TI - Long-term outcome of anatomical reconstruction versus tenodesis for the treatment of chronic anterolateral instability of the ankle joint: a multicenter study. AB - The long-term clinical outcome after anatomical reconstruction and tenodesis in the treatment of chronic anterolateral ankle instability was assessed in a retrospective multicentre study. The first group (AR) consisted of 25 patients (mean age at operation 22 yrs +/- 5.7) who underwent anatomical reconstruction and the second group (TE) of 29 patients (mean age 23 yrs +/- 6.6) who underwent tenodesis. For both groups, the mean follow-up period was 12.3 yrs (AR +/- 2.5 yrs, TE +/- 2.7 yrs). At physical examination, there were significantly more patients in the TE group (n=18) with a positive anterior drawer sign as compared with the AR group (n=7) (p=0.02). Medially located degenerative changes in the ankle joint as seen on standard radiographs were seen more often in the TE group (n=7) than in the AR group (n=1) (p=0.03). The mean talar tilt, 4.7 degrees in the AR group vs 6.9 degrees in the TE group, (p=0.02) and anterior talar translation, 2.9 mm in the AR group vs 4.3 mm in the TE group, (p=0.04) were significantly higher in the TE group at radiographic stress examination. According to the rating system developed by Good et al. (1975), significantly fewer patients in the TE group (n=8) had an excellent result as compared with the AR group (n=15) (p=0.03) and more patients in the TE group (n=9) had a fair or poor result (p=0.04) as compared with the AR group (n=2). We conclude that a tenodesis procedure does not restore the normal anatomy of the lateral ankle ligaments. When compared with anatomical reconstruction, a tenodesis leads to inferior results in terms of functional and mechanical stability, as well as overall satisfaction at long-term follow-up. PMID- 11428762 TI - When does the flat-top talus lesion occur in idiopathic clubfoot: evaluation with magnetic resonance imaging at three months of age. AB - Flat-top talus has been described as a pathologic change secondary to idiopathic clubfoot condition and/or as a direct result of nonoperative manipulation involving forced dorsiflexion and molding of the cartilaginous talus. No definitive study, however, on the etiology and the timing of the flat-top talus deformity has been performed to date. The authors evaluated the magnetic resonance images of eleven patients with idiopathic clubfoot deformities treated with 2 to 3 months of casting to assess if flattening of the talar dome occurred at this age with this amount of casting. All children were 3 months of age, were casted for a maximum of 2 to 3 months, and sedated before MRI examination. The images were evaluated for maximum talar head height, maximum talar body height, and deviation of the talar body from a perfect circle. Maximum talar head height ranged from 4 to 9 mm, maximum talar body height ranged from 6 to 10 mm. Eight of the eleven had maximum talar body measurements 3 to 5mm greater than maximum talar head height. Three of the eleven patients had head and body size of equal proportion. Two of the eleven had a talar body that was within 1 mm of a perfect circle. The remaining nine patients had perfectly round talar bodies. In the senior author's (RSD) experience with treating clubfeet, a substantial increase has been seen at operation in flat-top tali among children that were casted for more than 1 year before surgical correction, compared to children casted for 3 months before surgical correction. The current investigation indicated that although tali of children with clubfeet are abnormally shaped, the talar body remains larger than the talar head and maintains its roundness after two to three months of corrective casting. Maintenance of cast treatment for more than three months may lead to the flat-top talus deformity. The authors recommend surgical intervention following three months of failed manipulation and casting to prevent this deformity. PMID- 11428763 TI - Measurement of isolated subtalar range of motion: a cadaver study. AB - Fifteen fresh-frozen cadaveric lower extremities were studied to evaluate the reliability of measuring subtalar motion using a bubble inclinometer. There was high intra-observer reliability for manual inversion and eversion of the subtalar joint with the tibiotalar joint locked and unlocked. Poor correlation of radiographic and clinical measurements questioned the validity of bubble inclinometer measurements. The contribution of the tibiotalar joint to apparent subtalar motion, as measured clinically and radiographically, was found to be one third of the arc of motion, as compared to motion measured clinically and radiographically with the tibiotalar joint locked. PMID- 11428764 TI - The effect of heel elevation on strain within the plantar aponeurosis: in vitro study. AB - Mild, temporary reduction of symptoms from plantar fasciitis have been reported with the use of high heeled shoes (i.e. cowboy boots, ladies pumps). However, little is known on how heel elevation may contribute to a decrease in the pain and inflammation. The aim of this study was to quantify strain in the plantar aponeurosis in cadaveric feet with the use of various heel elevation configurations. An in vitro method that simulated "static" stance was used to determine the loading characteristics of the plantar aponeurosis (n = 12). Heel elevation was evaluated with blocks placed beneath the heel and with a contoured platform that simulated the arch profile of a shoe at three different heel heights (2.0, 4.0, 6.0 cm) with a level plane serving as the control. Strain in the plantar aponeurosis decreased with elevations of the heel that simulated the arch profile of a shoe at load levels (337, 450 N) (P < 0.05). Elevations of the heel with blocks did not significantly affect strain in the plantar aponeurosis (P < 0.05). Contrasting results of some specimen limbs compared with the overall means suggests that the influence of heel elevation on loading of the plantar aponeurosis may be dependent on individual variation and foot structure differences. Therefore, clinicians should be cautious in recommending heel elevation as a treatment for plantar fasciitis since some subjects may not achieve the desired decrease in plantar aponeurosis strain. PMID- 11428765 TI - Intra-articular neuropathic fracture of the calcaneal body treated by open reduction and subtalar arthrodesis. AB - A novel pattern of neuropathic intra-articular calcaneal fracture in a diabetic patient is described. This fracture combined proximal retraction of the tuberosity and body along with hindfoot collapse and plantar ulceration. Following control of the acute Charcot process with total contact casting, surgical reduction and subtalar arthrodesis was performed to stabilize the hindfoot and decrease the risk of recurrent ulceration. After healing, the patient successfully resumed ambulation and presently uses extra-depth shoes for daily activities. Despite the risks of surgical treatment, this difficult fracture may be treated operatively to maintain plantigrade alignment and offer limb salvage for the neuropathic patient. PMID- 11428766 TI - Speech after repair of isolated cleft palate and cleft lip and palate. AB - The speech of children with isolated cleft palate (CP) repaired by one surgeon has been compared with the speech of children with some form of unilateral cleft lip and palate (CLP) repaired by the same surgeon. All palate repairs included an intravelar veloplasty. We identified 57 children (5--12 years old) with cleft palates repaired in infancy, of which three patients with other medical problems were excluded. Of the 54 patients, 44 (81%) attended for review (27 CP, 17 CLP). Video recordings were analysed by two speech and language therapists, using the Cleft Audit Protocol for Speech. The CP patients had no evidence of permanent fistulas. Final speech outcomes were similar for CP and CLP patients. Intelligibility was normal in 10 (37%) CP and nine (53%) CLP patients. Mild consistent hypernasality was present in five (18.5%) CP and four (23.5%) CLP patients. No patients had moderate or severe hypernasality or nasal emission. Mild consistent hyponasality was present in five (18.5%) CP and five (29%) CLP patients. Moderate consistent hyponasality was present in one (4%) CP patient. Dysphonia was present in eight (30%) CP and seven (41%) CLP patients. Cleft-type characteristics were noted in 11 (41%) CP and nine (53%) CLP patients. No CLP patients but 10 (37%) CP patients had required a pharyngoplasty (P=0.004, Fisher's exact test). Possible reasons for this (age, cleft type, surgeon and surgery) are discussed. PMID- 11428767 TI - Cranial distraction osteogenesis in sheep using a totally implantable hydraulic mechanism: early results. AB - Conventional distraction osteogenesis has been performed by using either percutaneous pins attached to a thread and screw mechanism, or a thread and screw mechanism that is implanted but still requires turning by a percutaneous instrument or device. In these situations there is always the risk of infection passing to the bone via the percutaneous connection. We have developed a totally implantable hydraulic mechanism, which is activated in much the same way as injection into the port of a tissue expander. Preliminary in vitro testing has suggested that our mechanism can produce sufficient force at appropriate rates of distraction to have a promising role in distraction osteogenesis. We examined the effect of osseous distraction on the cranial vault. Expansion of the cranial vault by distraction osteogenesis is not widespread in clinical practice, in part because of the risk of infective complications of the extra-dural space. We were, therefore, presented with an ideal opportunity both to test the new distraction device and to examine the effect of distraction osteogenesis on the cranial vault. A sheep animal model was used in a pilot study to test the plausibility and examine the physiology of cranial-vault distraction osteogenesis using a totally implantable hydraulic device. Two sheep had a device implanted in the temporal fossa so as to push the craniotomised cranial cap upwards. Distraction was performed for 13 days beginning 1 week after craniotomy. After 9 weeks of consolidation, 5.5 mm and 7.4 mm of new woven bone of normal cranial thickness were demonstrated. PMID- 11428768 TI - Pre-expanded arterialised venous free flaps for burn contracture of the cervicofacial region. AB - Despite the fact that arterialised venous flaps provide thin good-quality tissue to repair defects of the face and neck, their clinical applications have been limited by an unstable postoperative course and variable flap necrosis. In an effort to resolve these problems, a tissue-expansion technique has been applied to the arterialised venous flap before flap transfer. Three pre-expanded arterialised venous free flaps have been used to treat post-burn scar contracture of the cervicofacial region. The donor site was confined to the forearm in each case. A rectangular expander was usually placed over the fascia of the flexor muscles in the proximal two-thirds of the forearm. The mean expansion period, volume and flap size were 44 days, 420 cm(3)and 147 cm(2), respectively. There were no complications caused by insertion and expansion. The cervicofacial region was successfully reconstructed, after excision of the post-burn contractures, with pre-expanded arterialised venous flaps, with no marginal necrosis or postoperative instability. Large thin arterialised venous flaps are well matched with the recipient defect in the cervicofacial area and the colour and texture match obtained with forearm tissue produced an aesthetically favourable result. Pre-expanded arterialised venous flaps are another new option for free flap reconstruction of the face and neck. PMID- 11428769 TI - Emergency treatment of accidental infusion leakage in the newborn: report of 14 cases. AB - Infusion leakage in the paediatric population of the intensive-care unit is known to cause skin necrosis and significant scarring around tendons, nerves and joints, extending the length of hospital stay. We report a series of 14 newborn children affected by accidental infusion leakage, and their early treatment with Gault's procedure: saline flush-out and liposuction. The results were good: there was no skin impairment in 11 cases and three cases of skin necrosis healed spontaneously. Early treatment of toxic infusion leakage in neonates is recommended to avoid skin necrosis at the site of extravasation. The two procedures proposed by Gault are simple and effective in such cases. They should be employed as early as possible in order to dilute and remove the toxin from the subcutaneous tissue. PMID- 11428770 TI - Decussation patterns of the platysma in Koreans. AB - The importance of the platysma muscle in plastic surgery is well known. During surgery, we have observed that the platysmal fibres do not merely decussate and interlace from each side but that sometimes one side of the muscle overlaps and covers the other side. The aim of this study was to clarify the precise anatomical variations in the decussation and overlapping patterns of the platysma muscle in 70 Korean cadavers. We defined three groups (types I--III) and four patterns (types A--D) according to the length of decussation and the pattern of overlapping, respectively. In 60 specimens (85.7%) we observed decussation and in 10 (14.3%) we did not. In type I there was 0--20 mm of decussating fibres below the mandibular border (43% of specimens). In type II, the decussation of fibres extended over more than 20 mm (43% of specimens). Type III had no decussation (14% of specimens). The proportions of type A (interlacing pattern), type B (right dominant), type C (left dominant) and type D (no decussation) were 41%, 29%, 16% and 14%, respectively. In 45% of the specimens, one side of the platysma covered and overlapped the other side. This is relevant to the removal of the subplatysmal fat in corset platysmaplasty, submental Z-plasty and vertical myectomy of the platysma muscle. PMID- 11428771 TI - Cranial-bone defects with depression deformity treated with ceramic implants and free-flap transfers. AB - Hydroxyapatite ceramic has many advantages in the treatment of cranial-bone defects. However, for large skull defects with severe depression deformities, it may be risky to use ceramic implants because an extradural dead space will be left and the overlying scalp will have to be closed under tension. In these cases microvascular free-flap transfers are a good solution. We have treated three patients for large skull defects with severe depression deformities after repeated local infections and several operations or irradiation. A latissimus dorsi myocutaneous flap was combined with a serratus anterior muscle flap to fill the extradural space under the ceramic implant in the first patient. A latissimus dorsi muscle flap was inserted under the ceramic implant in an irradiated site and a combined small serratus anterior muscle flap was used as a monitor in the second patient. A latissimus dorsi myocutaneous flap was used to cover the ceramic implant and fill the scalp defect in the third patient. The follow-up periods varied from 12 to 35 months (mean: 20.7 months). The clinical courses of all three patients were uneventful and no flap was lost. The extradural space can be reduced to some extent by making the ceramic implant slightly flatter or thicker, but in patients with severe depression deformities, whose brain expansion cannot be expected, a muscle flap should be transferred into the space. PMID- 11428772 TI - Sternomastoid-muscle transposition improves the cosmetic outcome of superficial parotidectomy. AB - A facial depressed deformity subsequent to superficial parotidectomy is unsightly. Although a facelift incision can improve the cosmetic outcome by concealing the scar, the hollow contour around the angle of the mandible remains conspicuous. We have attempted to mitigate this problem by transposition of the sternomastoid muscle. Transposition of the sternomastoid muscle to cover the parotid bed after superficial parotidectomy for benign tumour was performed in eight consecutive patients. The histopathology and postoperative results, including complications and patient satisfaction, were analysed. The depressed deformity was considerably alleviated in all eight patients. All patients except one, who had a wound infection, were satisfied with the cosmetic outcome postoperatively. The extra operative time required for sternomastoid-muscle transposition was only 10 min. There was no morbidity related to this additional procedure. By combining this simple method with a facelift incision, an appealing cosmetic outcome can be achieved after superficial parotidectomy. PMID- 11428773 TI - Endothelin receptor blockade improves oxygenation in contralateral TRAM flap tissue in pigs. AB - Partial skin and fat necrosis is the most common complication occurring in TRAM flaps. It is related to disturbances of the microcirculation and oxygenation in the contralateral part of the flap. It may be hypothesised that the development of necrosis is promoted by the vasoconstrictor endothelin, the production of which is enhanced in ischaemic flap tissues. The purpose of this study was to evaluate the effect of tezosentan, a new endothelin receptor blocker, on microcirculation and oxygenation in experimental TRAM flaps. The administration of tezosentan began preoperatively (3 mg/kg body weight) and then continued at a rate of 1.5 mg/kg/h. A TRAM flap with a skin island measuring 16 x 8 cm was raised in the middle of the epigastrium in minipigs. The flap was pedicled on the right superior epigastric vessels. Microcirculatory blood flow was measured with laser Doppler flowmetry and tissue oxygen tension was measured with a Clark-type microprobe. Dominant subcutaneous veins were cannulated in both the ipsilateral and the contralateral parts of the flap. Subdermal tissue oxygen tension in the contralateral part of the flap was significantly reduced 4h after surgery to 5 mmHg (ca. 48 mmHg in normal tissue) in the control group, but to only 12 mmHg in the group that had been administered tezosentan (P< 0.05). Furthermore, tezosentan significantly attenuated venous hypertension (14 mmHg versus 24 mmHg), as well as lactate (4.0 mmol/l versus 5.6 mmol/l) and haemoglobin (10.2 g/dl versus 11.4 g/dl) concentrations in the venous effluent of the contralateral part of the flap, although microcirculatory blood flow remained virtually unchanged. Our findings suggest that tezosentan improves oxygenation and metabolism in the jeopardised contralateral flap tissue, probably as a result of a decrease in venous vascular resistance and fluid extravasation. PMID- 11428774 TI - Anatomical relationship between arteries and veins in the scapular region. AB - Veins in the scapular region were investigated in five fresh cadavers, in which radiopaque materials were injected into both the arterial and the venous systems, to determine their locational relationship to the arteries. This radiographic technique is very useful for comparing veins and arteries. Many veins not accompanying arteries were observed, and veins in the skin and subcutaneous tissue were considered to consist of venae comitantes and non-venae-comitantes. The non-venae-comitantes formed a mainly polygonal venous network in the skin layer, and large communicating veins connected the venous network to the scapular or parascapular veins. These veins played a role in the drainage of venous blood that had perfused the dermis and, in this sense, they should be named cutaneous veins. PMID- 11428775 TI - Surgical technique and clinical application of the transverse gracilismyocutaneous free flap. AB - The unreliability of the distal skin component of the gracilis myocutaneous free flap has been frequently reported. To improve the reliability of the skin we orientated the cutaneous paddle in a transverse direction in the proximal third of the gracilis muscle, as first described by Yousif et al in 1993. Their anatomical studies showed that cutaneous branches of the dominant proximal pedicle have a pronounced tendency to travel in a transverse direction, supplying the skin anteriorly over the adductor longus and sartorius muscles and extending beyond the posterior margin of the gracilis muscle. We adopted this transverse design and transferred myocutaneous gracilis flaps measuring up to 17 x 9 cm. The transverse gracilis myocutaneous flap was dissected in the subfascial plane to include the peri-gracilis fascia, which preserved the fascial vascular network and thus optimised skin-paddle perfusion. Ten transverse gracilis myocutaneous free flaps were performed over 3 years. Skin paddles ranged in size from 10 x 7 cm (70 cm(2)) to 17 x 9 cm (153 cm(2)) with a mean of 113.4 cm(2). Five defects were located in the head and neck region, three in the lower leg, one in the thigh and one in the thorax. Patients were followed for an average of 16.6 months (range: 6--46 months). Minor complications (donor-site wound dehiscence and flap wound-edge separation) occurred in four patients;however, all 10 flaps survived and healed with complete cutaneous survival. PMID- 11428776 TI - The 'superficial' peroneal artery: a variation in cutaneous branching from the peroneal artery, nourishing the distal third of the leg. AB - Two different vascular patterns were encountered in cutaneous branches from the peroneal artery to the posterolateral aspect of the distal third of the leg in 22 flaps from 22 patients who underwent cutaneous or osteocutaneous peroneal flap surgery. In the type 1 vascular pattern, a branch from the peroneal artery, named the 'superficial peroneal artery', nourished the posterolateral aspect of the leg by splitting into several septocutaneous branches. In the type 2 pattern a few septocutaneous branches originated directly from the main peroneal artery and nourished the same area as that fed by the type 1 branch. The type 1 vascular pattern has not been reported to date but was seen in nine out of the 22 consecutive peroneal flaps. The superficial peroneal artery, with its considerable vascular diameter, may serve as a recipient vessel for free flaps or may serve as a donor nutrient vessel for a cutaneous flap, which can be transferred without sacrificing the main peroneal artery. Awareness of these two vascular patterns in the distal third of the leg should also help to reduce the small percentage of skin-flap failures that occur when the cutaneous or osteocutaneous peroneal flap is used. PMID- 11428777 TI - The use of a thin abdominal flap in degloving finger injuries. AB - Three multiple-finger degloving injuries were successfully resurfaced with thin abdominal flaps. This method offers immediate total wound coverage in multiple finger degloving injuries. PMID- 11428778 TI - The management of pressure sores by constant-tension approximation. AB - The management of pressure sores in medically compromised patients is problematic as they are seldom candidates for major surgical procedures. We treated 52 pressure sores in 49 patients (including three bilateral lesions) by constant tension approximation. The lesions were distributed as follows: 21 sacral, two ischial, 13 trochanteric and 16 posterior heel. The average age was 75 years and there were 22 males and 27 females. All patients had significant co-morbid conditions that, except for one wheelchair-bound person,confined them to bed. Nine patients were insensate; in two comatose cases this state was temporary. The management regime consisted of excision of the pressure sore and expansion of the skin and subcutaneous tissue using devices that, by means of continuous low-grade tension, approximated the wound edges, together with rigorous wound toilet and the judicious use of sutures. In total, 38 pressure sores healed primarily without recurrence, including three wounds that partially reopened and healed; 11 of these patients died, with healed wounds, at varying times between 2 and 48 months after treatment. Nine patients whose lesions were closed died within a month of treatment, and five patients, three of whose lesions were completely closed and two partially closed, were lost to follow-up; their lesions were not regarded as healed. This minimally invasive technique used to close pressure sores in elderly patients with co-morbid conditions has a satisfactory success rate. PMID- 11428779 TI - Development of a bullous pemphigoid after split-skin grafting. AB - An elderly female patient developed blisters localised to her split-skin donor and graft sites. Despite repeated swabs, bacteriology and virology returned negative; multiple antibiotic courses did not improve her condition, which generalised within weeks. Histology and immunofluorescence confirmed a clinical diagnosis of bullous pemphigoid and her condition resolved rapidly after treatment with prednisolone. Bullous pemphigoid can be precipitated or exacerbated by surgery and the diagnosis should be considered when patients present with persistent localised or generalised blisters, especially when microbiological investigations are negative. PMID- 11428780 TI - Sensory and motor function impairment after brachial plexuscord compression by a pedicled latissimus dorsi flap. AB - A case of neuropraxia of the sensory and motor nerve fibres of the brachial plexus is reported after successful transfer of an ipsilateral pedicled myocutaneous latissimus dorsi flap to reconstruct a large-volume tissue defect in the neck resulting from a shotgun injury. PMID- 11428781 TI - Use of bilateral retroangular flaps for reconstruction of the glabella and nose. AB - We report the use of bilateral retroangular flaps for reconstruction of the glabella and nasal tip following extirpation of a cavernous haemangioma. The vascularity of the flaps was good, and the colour and texture match excellent. Donor-site closure was easy, and the scar in the nasolabial groove was not conspicuous. Our experience demonstrates that this flap is useful for reconstruction of skin defects of not only the nose but also the glabella and forehead. By raising bilateral flaps, a wide skin defect in the centre of the face can be repaired. PMID- 11428782 TI - Acute facial-nerve paralysis with parotid adenolymphoma. AB - An 82-year-old woman presented with acute facial paralysis in association with parotid swelling following fine-needle aspiration cytology of a parotid adenolymphoma. Evacuation of a tense haematoma decompressed the nerve and led to complete recovery within 6 months. Additionally, the tumour is no longer evident, either clinically or radiologically, and fine-needle aspiration cytology appears to have been both diagnostic and, inadvertently, therapeutic in this case. PMID- 11428783 TI - Subcranial anterior skull base dural repair with galeal frontalis flap. AB - The pericranial flap has been widely used to repair surgically created anterior skull base defects during craniofacial resection. This report describes a technique of subcranial dural repair with a galeal frontalis flap after resection of an ethmoidal adenoid cystic carcinoma. PMID- 11428784 TI - Aggressive keloid scarring of the Caucasian wrist and palm. AB - Keloid scarring of the distal upper extremity is very rare. We report a Caucasian woman who presented with aggressive keloids of the hand and wrist causing De Quervain's syndrome, superficial radial-nerve entrapment and ulnar-nerve compression at the wrist. Multiple operations were required to alleviate her symptoms. A number of management conundrums arose, requiring defensive planning to pre-empt the possible complications of recurrent keloid scarring as a result of the surgical procedures. PMID- 11428785 TI - Use of ovum forceps to hold the ear lobule. PMID- 11428786 TI - Self-examination for metastatic melanoma is a simple and worthwhile strategy. PMID- 11428788 TI - Free-flap debulking: the safe option. PMID- 11428789 TI - A technical refinement for removal of a ganglion. PMID- 11428790 TI - Osmotic instability of hydrogel-filled breast implants. PMID- 11428791 TI - Further comment on Trilucent breast implants. PMID- 11428792 TI - Hands off! PMID- 11428794 TI - Mr angiography in cerebrovascular disease. AB - MR angiography has become a realistic diagnostic option for patients with neurovascular disease. MR angiography is not a single imaging sequence, but a collection of related methods for obtaining angiographic data. As a guide for practice, we review the literature on MR angiography in a spectrum of neurovascular indications with particular attention paid to choice of technique. The principles underlying the different techniques available are also presented. Summers, P. E.et al. PMID- 11428795 TI - Thermal injuries associated with MRI. AB - Most physicians are aware of the absolute contraindications to magnetic resonance imaging (MRI). However, less familiar is the potential for an MRI-induced thermal or electrical burn associated with electrical monitoring devices. Although detailed studies concerning the burn hazard in MRI have not been reported, it is widely believed that direct electromagnetic induction in looped cables associated with the patient is responsible for the excessive heating and it is on this theory that present guidelines are based. Recent reports have however indicated that other mechanisms may cause the heating of metal, either in or on the patient. This document reviews numerous reported burn injuries sustained during MRI and addresses the underlying heating mechanisms possibly causing these events. PMID- 11428796 TI - Accuracy and reliability of quantitative measurements in coronary arteries by multi-slice computed tomography: experimental and initial clinical results. AB - AIM: To evaluate the accuracy of non-invasive measurements within coronary arteries by multi-slice computed tomography (MSCT). We present experimental as well as clinical data. MATERIALS AND METHODS: Silicon tubes simulating coronary arteries (outer diameter 6 mm, lumen diameter within stenotic area 2 mm) were used for experimental studies. Clinical data were derived from 15 patients in whom vessel diameters were assessed by MSCT, intracoronary ultrasound (ICUS) and quantitative coronary angiography (QCA). MSCT were performed in a Somatom Volume Zoom(trade mark)CT system (Siemens, Forchheim, Germany) at 2 collimated slice widths (2.5 mm, 1.0 mm). RESULTS: Outer silicon tube diameters were overestimated by MSCT (6.56 mm +/- 0.32 mm). All measurements revealed significantly better results on 1.0 collimation compared to 2.5 mm collimation (outer diameter: 6.36 mm +/- 0.22 mm vs 6.76 mm +/- 0.27 mm, P < 0.0001; lumen diameters: 1.83 mm +/- 0.14 mm vs 1.51 mm +/- 0.19 mm, P < 0.0001). The comparison of vessel diameters within human coronary arteries revealed comparable results between ICUS and MSCT (4.89 mm +/- 0.67 mm vs 4.91 mm +/- 0.71 mm, P = 0.79, r = 0.79, P < 0.0001). QCA measurements showed significantly lower results (3.67 +/- 0.71, P < 0.0001, r = 0.62, P < 0.001). CONCLUSIONS: Experimental as well as initial clinical results indicate acceptable reliability and accuracy of quantitative measurements by MSCT, when using thin collimated slice widths. Partial volume effects lead to a systematic overestimation of vessel size. MSCT has the potential to become an important non-invasive diagnostic tool in patients with coronary artery disease. PMID- 11428797 TI - Spinal subdural haematoma: a study of three cases. AB - AIMS: To define the clinical, computed tomography (CT) and magnetic resonance imaging (MRI) features and the role of MRI in the follow-up of spinal subdural haematoma (SSH), and to compare these findings with those of spinal epidural haematomas (SEH). METHODS: We report three cases of SSH (two women, one male, age: 50-74 years). Two patients were on anticoagulant therapy; in the other case the SSH was spontaneous. All the patients were examined 1-3 days after the onset of the symptoms. All of them had CT, two had MRI and one had angiography. Two patients underwent surgery. RESULTS: The haematoma was located in the thoracolumbar region (two) and in the thoracic region (one), extending from five (two) to 11 vertebral body levels (one). The haematomas were posteriorly located with lateral extension. The transverse shape differed with the level: biconvex, biloculated or circumferential. The haematomas were hyperdense on CT. On MRI, SSH yielded high signal on both T1 and T2. The integrity of the posterior fat pads, which was well shown on CT and MRI, and the visualization of the dura mater demonstrated the intradural location of these collections, making them easily distinguishable from spinal epidural haematoma. MRI provides better evaluation of the longitudinal extent. Our results are compared with those reported in the literature. CONCLUSION: MRI is superior to CT for diagnosis and follow-up of SSH. Our findings and those reported in the literature show that the MR features of SSH are quite specific and allow differentiation from SEH. PMID- 11428798 TI - Erdheim-chester disease. AB - Erdheim-Chester disease (ECD) is a rare histiocytic infiltrative disease, characterized by long bone sclerosis. We describe two confirmed cases of ECD, both of which demonstrate non-malignant retroperitoneal and peri-renal infiltration causing dilatation of the upper renal tracts. The cases are illustrated with contrast studies, computed tomography (CT) and magnetic resonance imaging (MRI). Typical sclerosis of the long bones was apparent on radiography. Both cases have been treated conservatively to date.A brief review of the literature regarding the manifestations of ECD is included. In cases of non-malignant retroperitoneal infiltration, ECD should be considered as a diagnosis and radiographs of the long bones performed. PMID- 11428799 TI - MRI findings in neuro-behcet's disease. AB - AIM: To evaluate the pattern and site of involvement in neuro-Behcet's disease (NBD). MATERIALS AND METHODS: Twenty-one patients with NBD were evaluated. Using 1.5T magnetic resonance imaging (MRI), T1-weighted axial and sagittal images, gadolinium enhanced axial and coronal images and T2-weighted axial images were obtained. RESULTS: The brainstem, basal ganglia, cerebral white matter, internal capsule, thalamus and spinal cord were involved in eighteen, nine, nine, seven, six and two patients, respectively. In nine patients with cerebral white matter involvement, four had subcortical involvement and three had periventricular involvement, in addition to two patients with focal deep white matter lesions. Among the brainstem lesions, pons involvement was seen in fourteen patients, all had ventrally located lesions, and nine had tegmental involvement. Midbrain involvement was seen in fourteen patients; the cerebral peduncle was involved in 11 of these. Five patients had brainstem atrophy: two cases were demonstrated at initial MRI, the other three cases were seen on follow-up MRI. Pyramidal signs, the most common neurological signs, were demonstrated in fourteen patients. Follow-up MRI was obtained 10 days to 20 months after the initial MRI in eight cases; all showed changes in size, shape and site of involvement. After gadolinium enhancement, thirteen patients demonstrated mottled non-confluent enhancement in the brainstem (eight patients), posterior limb of the internal capsule (three patients), pachymeninges (two patients) and spinal cord (two patients). CONCLUSION: NBD manifests a reversible course, but chronic NBD may result in brainstem atrophy. Characteristic involvement along the corticospinal tract is well correlated with neurological signs. PMID- 11428800 TI - An in-vitro study to understand successful free-hand ultrasound guided intervention. AB - AIMS: To describe the optimum conditions for successful free-hand ultrasound guided intervention using an in-vitro experiment. MATERIALS AND METHODS: Optimal free-hand biopsy conditions were determined using targets buried in a purpose built interventional phantom. The needle/probe distance and probe/phantom surface angle were changed whilst recording the needle track lengths and needle conspicuity. RESULTS: Optimal free-hand conditions included a needle/probe distance of 2-3 cm in combination with a needle/probe angle 55-60 degrees. This enabled excellent needle conspicuity with short needle track length and early needle tip visualization. CONCLUSION: Understanding how to optimize the free-hand conditions will help the interventionalist to obtain greater performance using a variety of puncture devices. This concept is likely to be particularly important to the novice interventionalist as well as to those teaching the free-hand technique. PMID- 11428801 TI - In-vitro visualization of biopsy needles with ultrasound: a comparative study of standard and echogenic needles using an ultrasound phantom. AB - AIM: To demonstrate the circumstances where echogenically enhanced biopsy needles confer an increase in visualization during ultrasound-guided biopsy using an in vitro experiment. MATERIALS AND METHODS: An experiment using both standard and echogenically enhanced needles was conducted in a purpose built interventional phantom. The needle tip echogenicity and shaft visualization was recorded at different angles (20 and 60 degrees ) between the ultrasound probe and needle. The needle tip echogenicity was also recorded in four different angles of bevel rotation. The experiment was performed using a total of 18 different needles of 18 and 20 gauge. RESULTS: The experiment confirmed some previous findings. The needle was better visualised at an angle of approximately 60 degrees to the probe. The needle tip was optimally visualized with the bevel facing the transducer or 180 degrees to it. Needle tip movement improved conspicuity. Using an optimal angle of approach (60 degrees ) the use of an echogenic needle did not confer any increase in conspicuity. If the transducer to needle angle was suboptimal (20 degrees ) then there was increased visualization of some echogenically enhanced needles. CONCLUSION: Optimisation of the conditions prior to ultrasound-guided needle biopsy will increase the visualization of the chosen device thus aiding the interventionalist. Needle gauge, bevel position, movement of the needle and the probe to needle angle all affect conspicuity. Needle coating or roughening does not improve tip visualization if the angle between the needle and probe can be optimized (60 degrees ). In unfavourable situations when the angle is suboptimal (20 degrees ), we have shown that echogenic enhancement of the needle tip can make it more visible. PMID- 11428802 TI - Radiological management of an inoperable cholangiocarcinoma arising from a choledochal cyst. PMID- 11428803 TI - Hepatic tumours in glycogen storage disease type 1 (von Gierke's disease). PMID- 11428804 TI - Choledochoduodenal fistula complicating non-hodgkin's lymphoma of the duodenum during chemotherapy. PMID- 11428805 TI - Posterior interosseous nerve palsy caused by synovial chondromatosis of the elbow joint. PMID- 11428806 TI - CT angiographic appearances of carotico-cavernous fistula. PMID- 11428807 TI - Cine magnetic resonance imaging of isolated right ventricular outflow obstruction in hypertrophic cardiomyopathy. PMID- 11428808 TI - Diabetic patients and acute coronary syndromes. PMID- 11428809 TI - Gaps and opportunities in the practice of medicine: the need for improved systems of care. PMID- 11428810 TI - Every piece is important to build a puzzle. What can we learn from early experiences with balloon PTCA? PMID- 11428811 TI - Importance of control of fluid volumes in heart failure. PMID- 11428812 TI - Do heart transplant recipients need annual coronary angiography? PMID- 11428813 TI - The general concepts of an equivalence trial, applied to ASSENT-2, a large-scale mortality study comparing two fibrinolytic agents in acute myocardial infarction. PMID- 11428814 TI - Angiogenesis and the ischaemic heart. PMID- 11428815 TI - At what coronary risk level is it cost-effective to initiate cholesterol lowering drug treatment in primary prevention? AB - BACKGROUND: The entire risk factor profile should be taken into account when considering initiating cholesterol lowering drug treatment. Recent treatment guidelines are therefore based on the absolute risk of coronary heart disease. We estimated at what coronary risk it is cost-effective to initiate cholesterol lowering drug treatment in primary prevention for men and women of different ages in Sweden. METHODS: The cost-effectiveness was estimated as the incremental cost per quality-adjusted life-year (QALY) gained of cholesterol lowering drug treatment. Treatment was assumed to lower the risk of coronary heart disease by 31%. The analysis was carried out from a societal perspective including both direct and indirect costs of the intervention and morbidity, and the full future costs of decreased mortality. The coronary risk, in a Markov model of coronary heart disease, was raised until the cost per QALY gained corresponded to a specific threshold value per QALY gained. Three different threshold values were used: $40,000, $60,000 and $100,000 per QALY gained. RESULTS: The risk cut-off value for when treatment is cost-effective varied with age and gender. If society is willing to pay $60,000 to gain a QALY it was cost-effective to initiate treatment if the 5-year-risk of coronary heart disease exceeded 2.4% for 35-year old men, 4.6% for 50-year-old men, and 10.4% for 70-year-old men. The corresponding risk cut-off values for women were 2.0%, 3.5% and 9.1%. CONCLUSIONS: The results can serve as a basis for treatment guidelines based on cost-effectiveness. PMID- 11428816 TI - Siblings of myocardial infarction patients are overlooked in primary prevention of cardiovascular disease. AB - AIMS: Patients with arterial hypertension or hypercholesterolaemia may benefit from medical therapy for primary prevention of myocardial infarction. Preventive therapy may be particularly effective in individuals with a positive family history for myocardial infarction since such subjects are at high risk for coronary events. The objective of this population-based study was to analyse the risk profile as well as the current utilization of preventive strategies in asymptomatic siblings of patients with myocardial infarction. METHODS AND RESULTS: We studied siblings of 325 patients with premature myocardial infarction from the Augsburg MONICA myocardial infarction registry by standardized questionnaire, blood pressure recordings, and biochemical measurements. Out of 580 siblings, 510 were free of coronary heart disease symptoms. With multiple risk factors being present in most asymptomatic siblings, 29.4% of asymptomatic individuals had an estimated individual 10-year risk for a major cardiovascular event of > or = 20%, or when projected to the age of 60. According to the guidelines of the European Societies of Cardiology, Atherosclerosis, and Hypertension (ESC/EAS/ESH) from 1994 (1998 guidelines in parenthesis) dietary and lifestyle interventions were indicated for arterial hypertension in 48.1% (43.0%) and/or for hypercholesterolaemia in 17.3% (78.8%). Drug treatment was indicated for arterial hypertension in 27.9% (30.6%) and for hypercholesterolaemia in 13.6% (19.1%) of asymptomatic siblings. Of those individuals with the respective indication, actual drug treatment was given for arterial hypertension in 91.5% (83.3%) and for hypercholesterolaemia in 46.4% (33.0%). However, treatment targets were reached in only 31.0% (28.2%) with arterial hypertension and in 7.2% (5.2%) with hypercholesterolaemia, respectively. CONCLUSIONS: Most asymptomatic individuals with positive fraternal family history have more than one modifiable risk factor. Interestingly, a large number of these individuals appears to be under medical surveillance as many receive some sort of drug treatment. However, this therapy did not meet the treatment goals in the majority of those with arterial hypertension and/or hypercholesterolaemia. Thus, although individuals with a positive fraternal history for myocardial infarction can be easily identified, implementation of sufficient preventive strategies continues to be poor in a Western European country. PMID- 11428817 TI - Long-term clinical outcome after coronary balloon angioplasty: identification of a population at low risk of recurrent events during 17 years of follow-up. AB - AIMS: This study reports the clinical outcome, up to 17 years, of the first 856 consecutive patients treated by coronary angioplasty at a single centre and attempts to identify a subgroup of patients at low risk of adverse events. METHODS AND RESULTS: Follow-up status was established via hospital and general practitioner records and the civil registry. Median follow-up was 16 years. The overall 5-, 10-, 15- and 17-year survival was 90%, 78%, 64% and 58%, respectively and corresponding event-free survival was 53%, 33%, 22% and 19%. After 32% of patients had experienced a major adverse cardiac event in the first year, the annual coronary re-intervention incidence thereafter and, even beyond year 10, remained at 2%--3%. Using multivariable Cox regression, significant independent predictors of mortality were advanced age, diabetes, multivessel disease and impaired left ventricular function at the time of PTCA. A subgroup of 26% of the patients with none of these risk factors had a survival rate similar to the general Dutch population matched for age and gender (at 5 years: 96%, at 10 years: 89% and at 15 years: 83%). CONCLUSION: Although the majority of patients (>80%) experienced a further cardiac event during the 17 years after their first angioplasty procedure, in those non-diabetics under 60 years with single-vessel disease and good left ventricular function, prognosis was similar to the general population. PMID- 11428818 TI - Continuous ambulatory monitoring of absolute right ventricular pressure and mixed venous oxygen saturation in patients with heart failure using an implantable haemodynamic monitor: results of a 1 year multicentre feasibility study. AB - BACKGROUND: Implantable sensors that monitor haemodynamics over time may be useful in patients with heart failure. This multicentre study assessed the feasibility of a system that has one sensor measuring absolute pressure and another measuring mixed venous oxygen saturation (SvO(2)). Both sensors were mounted on leads that were implanted in the right ventricle. METHODS: Twenty-one patients with heart failure (NYHA II-III) were included. Comparisons were made to right heart catheterizations at implant and at 2, 6 and 12 months thereafter. Patients underwent several haemodynamic provocations during the catheterizations. RESULTS: Overall, among functioning sensors, the IHM-1 values were highly correlated with reference values for all time points during all provocations, demonstrating high reproducibility and stability (r(2)=0.91, 0.79 and 0.78 for systolic, right ventricular diastolic and SvO(2), respectively). Although IHM-1 underestimated reference pressure values by 4.5 mmHg and SvO(2)by 1.6%, this difference was consistent across provocation and stable over 12 months of follow up. Twelve of the 21 oxygen sensors failed to function and two pressure sensors had component failures. Preliminary analysis of long-term data revealed haemodynamic patterns that may be key indicators for therapeutic interventions. CONCLUSION: This multicentre feasibility study demonstrated the accuracy and stability of sensors implanted in the right ventricle. The IHM-1, using right ventricular pressures and SvO(2), with improved performance, might be useful in the study of pathophysiological mechanisms and treatment interventions in heart failure. PMID- 11428819 TI - Norepinephrine induces action potential prolongation and early afterdepolarizations in ventricular myocytes isolated from human end-stage failing hearts. AB - AIMS: Congestive heart failure is characterized by high levels of norepinephrine which is considered to be arrhythmogenic. It is unclear whether increased norepinephrine is only a marker of the severity of heart failure or whether it directly triggers ventricular arrhythmias. METHODS AND RESULTS: Ventricular myocytes were isolated from eight explanted hearts of patients with end-stage heart failure (ischaemic or dilated cardiomyopathy). With the whole-cell configuration of the patch-clamp technique the effect of 1 micromol x l( 1)norepinephrine on action potentials and membrane currents was studied. The cells had a membrane capacitance of 256 +/- 25 pF (n = 26) and action potential duration (APD90) during control conditions was 620 +/- 45 ms at 1 Hz (n = 14). Norepinephrine induced action potential prolongation in all cells and early afterdepolarizations in 50% of them. Norepinephrine significantly increased the calcium current but had no effect on the delayed rectifier current, the inward rectifier current or the transient outward current. Norepinephrine also significantly increased the steady-state calcium window-current measured between 40 and 0 mV. CONCLUSIONS: In contrast to many animal species, norepinephrine induces action potential prolongation in ventricular myocytes from human failing hearts, as well as early afterdepolarization, by an increase in both the calcium peak current and window current. Thus norepinephrine seems to be an important arrhythmogenic factor in congestive heart failure. PMID- 11428820 TI - Resting echocardiography and quantitative dipyridamole technetium-99m sestamibi tomography in the identification of cardiac allograft vasculopathy and the prediction of long-term prognosis after heart transplantation. AB - AIMS: To evaluate the accuracy of echocardiography in conjunction with quantitative high-dose dipyridamole technetium-99m sestamibi tomography (SPECT) in detecting coronary allograft vasculopathy. METHODS AND RESULTS: Seventy-eight consecutive heart transplant recipients underwent echocardiography while at rest and high-dose dipyridamole SPECT within 48 h of a yearly angiogram. Resting wall motion abnormalities were considered significant if present in two or more segments. SPECT was considered abnormal in the presence of reversible/fixed defects. The coronary angiogram was normal in 53, showed non-significant coronary allograft vasculopathy in 13 and significant (> or = 50% stenosis) coronary allograft vasculopathy in 12 cases. Resting wall motion abnormalities were observed in nine cases and perfusion defects in 20. Echocardiography and SPECT were concordant in 59 cases (five positive and 54 negative); in these, accuracy was 100% for significant coronary allograft vasculopathy and 83% for any coronary allograft vasculopathy. Over 6.5+/-2 years, 17 patients suffered coronary allograft vasculopathy-related events, including death in six and retransplantation in three. Resting wall motion abnormalities, SPECT perfusion defects and angiographic coronary allograft vasculopathy were significant predictors of cardiac events. CONCLUSION: Normal resting wall motion at echocardiography coupled to normal stress myocardial perfusion, rules out the presence of significant coronary allograft vasculopathy in many heart transplant recipients. Conversely, resting wall motion abnormalities and perfusion defects strongly predict cardiac events. Therefore, a strategy which reserves angiography for patients with resting wall motion abnormalities and/or perfusion defects may be safe and cost-effective. PMID- 11428822 TI - Is a mechanical or a metabolic approach superior in the treatment of coronary disease? Results of the atorvastatin versus revascularization (AVERT) trial. PMID- 11428823 TI - About EMIP-FR and reperfusion damage in AMI: a comment to the comment. PMID- 11428824 TI - The EMIP-FR Study: the evolution of scientific background as a non-controlled parameter. PMID- 11428825 TI - Trimetazidine and reperfusion injury. PMID- 11428826 TI - Trimetazidine in AMI. PMID- 11428829 TI - Are the data on trends in case-fatality in patients with acute myocardial infarction observed in Scotland applicable across Europe? PMID- 11428830 TI - The Q wave infarct; phenomenon or chimera? PMID- 11428831 TI - 'Localized' right ventricular morphological abnormalities in patients with the Brugada syndrome: what is their significance? PMID- 11428832 TI - Thromboprophylaxis for atrial flutter. PMID- 11428833 TI - D-transposition of the great arteries: post-operative evaluation by breath-by breath analysis of ventilation and pulmonary gas exchange during exercise. PMID- 11428834 TI - Cost-effectiveness of implantable cardioverter-defibrillators. PMID- 11428835 TI - The role of C-type natriuretic peptide in cardiovascular medicine. PMID- 11428836 TI - Expression of RGS3, RGS4 and Gi alpha 2 in acutely failing donor hearts and end stage heart failure. AB - BACKGROUND: Regulators of G-protein Signalling (RGS) proteins have been shown to limit in vitro signalling of G proteins. In common with end-stage heart failure, we have recently shown that upregulation of the inhibitory G-protein, Gialpha, occurs in acutely failing donor hearts unused for transplantation due to severe myocardial dysfunction. In light of recent data on RGS proteins, we have evaluated mRNA and protein expression of RGS3, RGS4 and Gialpha2 in the myocardium from normal, end-stage failing and acutely failing unused donor hearts. METHODS AND RESULTS: Myocardial samples were obtained from end-stage failing hearts explanted prior to transplantation (n=19), unused donor hearts with ejection fractions < 30% (n=14) and used donor hearts with good function (ejection fraction > 60%) (n=4-7). mRNA levels were quantified using quantitative reverse transcriptase polymerase chain reaction. Levels of RGS3 and RGS4 mRNA were found to be significantly upregulated in unused donor and end-stage failing myocardium (P < 0.05 and 0.01, and P < 0.05 and 0.02, respectively) compared to non-failing hearts. Protein abundance of RGS3 and RGS4 was found to be higher in myocardium from end-stage failing hearts, and relative RGS4 expression higher in unused donor hearts. CONCLUSIONS: We show here that RGS3 and RGS4 mRNA and protein expression is upregulated in human heart failure. These observations suggest that RGS4 may be induced in the heart to regulate cell signalling pathways in response to hypertrophy, and support the existence of a negative feedback loop for the long-term regulation of hypertrophy. PMID- 11428837 TI - Reduced costs with bisoprolol treatment for heart failure: an economic analysis of the second Cardiac Insufficiency Bisoprolol Study (CIBIS-II). AB - BACKGROUND: Beta-blockers, used as an adjunctive to diuretics, digoxin and angiotensin converting enzyme inhibitors, improve survival in chronic heart failure. We report a prospectively planned economic analysis of the cost of adjunctive beta-blocker therapy in the second Cardiac Insufficiency BIsoprolol Study (CIBIS II). METHODS: Resource utilization data (drug therapy, number of hospital admissions, length of hospital stay, ward type) were collected prospectively in all patients in CIBIS II. These data were used to determine the additional direct costs incurred, and savings made, with bisoprolol therapy. As well as the cost of the drug, additional costs related to bisoprolol therapy were added to cover the supervision of treatment initiation and titration (four outpatient clinic/office visits). Per diem (hospital bed day) costings were carried out for France, Germany and the U.K. Diagnosis related group costings were performed for France and the U.K. Our analyses took the perspective of a third party payer in France and Germany and the National Health Service in the U.K. RESULTS: Overall, fewer patients were hospitalized in the bisoprolol group, there were fewer hospital admissions per patient hospitalized, fewer hospital admissions overall, fewer days spent in hospital and fewer days spent in the most expensive type of ward. As a consequence the cost of care in the bisoprolol group was 5-10% less in all three countries, in the per diem analysis, even taking into account the cost of bisoprolol and the extra initiation/up-titration visits. The cost per patient treated in the placebo and bisoprolol groups was FF35 009 vs FF31 762 in France, DM11 563 vs DM10 784 in Germany and pound4987 vs pound4722 in the U.K. The diagnosis related group analysis gave similar results. INTERPRETATION: Not only did bisoprolol increase survival and reduce hospital admissions in CIBIS II, it also cut the cost of care in so doing. This 'win-win' situation of positive health benefits associated with cost savings is favourable from the point of view of both the patient and health care systems. These findings add further support for the use of beta-blockers in chronic heart failure. PMID- 11428838 TI - Localized right ventricular morphological abnormalities detected by electron-beam computed tomography represent arrhythmogenic substrates in patients with the Brugada syndrome. AB - AIMS: This study was designed to determine, using electron-beam CT, whether there are morphological abnormalities in patients with the Brugada syndrome and to elucidate the relationship between those abnormalities and arrhythmogenesis. METHODS AND RESULTS: Twenty-six consecutive patients with the Brugada syndrome and 23 age- and gender-matched control subjects (controls) were evaluated for morphological abnormalities using electron beam CT. Electron beam CT demonstrated morphological abnormalities of the right ventricle in 21 (81%) of 26 patients, but in only two (9%) of 23 controls. The sites of morphological abnormalities were the right ventricular outflow tract area in 17 patients and the inferior wall of the right ventricle in four patients. Of the seven patients with monoform premature ventricular contractions recorded only in the acute phase, four of the five patients with premature ventricular contractions from the right ventricular outflow tract area had morphological abnormalities in the right ventricular outflow tract area, and the other two patients with premature ventricular contractions from the inferior wall of the right ventricle had morphological abnormalities in the inferior wall of the right ventricle. CONCLUSION: The sites of morphological abnormalities detected by electron beam CT in patients with the Brugada syndrome were related to the origins of premature ventricular contractions recorded only in the acute phase, which may trigger ventricular fibrillation. These morphological abnormalities may be related to arrhythmogenic substrates in patients with the Brugada syndrome. PMID- 11428839 TI - Thromboembolic risk in atrial flutter. The FLASIEC (FLutter Atriale Societa Italiana di Ecografia Cardiovascolare) multicentre study. AB - AIMS: Patients with atrial flutter are believed to be at lower risk of thromboembolism than patients with atrial fibrillation. However, the incidence of atrial thrombi and the need for anticoagulation in patients with atrial flutter is not well established. METHODS AND RESULTS: A prospective observational multicentre study was undertaken to assess the frequency of atrial thrombi and spontaneous echocontrast and the prevalence for aortic complex atherosclerotic lesions in a cohort of unselected patients with atrial flutter. We evaluated 134 patients (102 male, aged 70+/-9 years); exclusion criteria were history of atrial fibrillation, rheumatic mitral valve disease and mitral mechanical prosthesis. The median of atrial flutter duration was 33 days. Twelve patients had been taking warfarin for more than 7 days. One hundred and twenty-four patients (94%) underwent a transoesophageal echocardiogram, which revealed left atrial appendage thrombi in two patients (1.6%) and right atrial thrombi in one patient (1%). At least moderate left atrial echocontrast was found in 16/124 patients (13%). Complex atherosclerotic aortic plaques were detected in 10 patients (8%). Atrial flutter conversion was attempted in 93/134 patients (69%). At the 1-month follow up, two patients experienced a thromboembolic event following restoration of sinus rhythm. CONCLUSIONS: Atrial thrombi and echocontrast, and complex aortic atherosclerotic plaques are relatively uncommon in patients with atrial flutter. Post-cardioversion embolism was observed in two patients in our study population. PMID- 11428840 TI - Cardiorespiratory exercise function after the arterial switch operation for transposition of the great arteries. AB - AIMS: This study compares the functional outcome and cardiorespiratory response to exercise, in patients who have undergone arterial switch for transposition of the great arteries, with normal controls and patients who have undergone atrial switch operation. METHODS AND RESULTS: Fifteen patients who had undergone arterial switch (mean age 8.5+/-2.9 years) were compared to 32 patients who had undergone atrial switch (9.2+/-1.8 years) and 27 normal controls (8.5+/-2.1 years). Exercise testing was performed on a treadmill and gas exchange measured breath-by-breath. Aerobic capacity, assessed by determination of the ventilatory anaerobic threshold, averaged 91+/-7.8% of normal (95% confidence limits: 92 108%) for arterial switch and 75.1+/-13.1% for atrial switch (P<0.001 patients vs normals). Aerobic exercise function was evaluated by calculation of the slope of oxygen uptake vs exercise intensity. The mean value for this slope was 2.0+/-0.25 for arterial switch, 2.5+/-0.46 for normals and 1.7+/-0.80 for atrial switch (P<0.05; patients vs normals). Efficiency of the pulmonary gas exchange was assessed by calculation of the slope of ventilation vs carbon dioxide output during exercise. This averaged 38.7+/-14.7 for arterial switch, 48.1+/-14.1 for atrial switch and 30.3+/-7.6 for normals (P<0.001; patients vs normals). CONCLUSION: Cardiorespiratory exercise function is at, or slightly below, the lower limit of normal in patients with arterial switch, while the lowest values were observed for those who had undergone atrial switch. PMID- 11428842 TI - Blood glucose in the CCU: time to measure. PMID- 11428843 TI - Secondary prevention: is inpatient rehabilitation effective or are lifestyle changes to be replaced by drugs? PMID- 11428844 TI - Statins and cardiovascular disease--major therapeutic advances but are we seizing the moment? PMID- 11428845 TI - Fibrinolysis and stenting in acute myocardial infarction: newlyweds destined for a 'menage a trois'? PMID- 11428846 TI - A sympathetic heart and the art of dying. PMID- 11428847 TI - Wearing your heart in your sleeve? PMID- 11428848 TI - Indications for implantable cardioverter defibrillator (ICD) therapy. Study Group on Guidelines on ICDs of the Working Group on Arrhythmias and the Working Group on Cardiac Pacing of the European Society of Cardiology. PMID- 11428849 TI - Social and psychosocial influences on sudden cardiac death, ventricular arrhythmia and cardiac autonomic function. PMID- 11428850 TI - Myocardial infarction and prevalence of diabetes mellitus. Is increased casual blood glucose at admission a reliable criterion for the diagnosis of diabetes? AB - AIMS: To investigate the prevalence of diabetes mellitus in patients with acute myocardial infarction and to determine whether casual blood glucose and haemoglobin A1c measured at admission could be used to diagnose diabetes mellitus. METHODS AND RESULTS: A prospective study that included all patients with acute myocardial infarction hospitalized during a one-year period at a coronary care unit. Casual blood glucose was measured at admission, fasting blood glucose during the hospital stay and, if necessary for classification, 2-3 months after discharge. Haemoglobin A1c was measured once at admission. Of 305 patients included in the study, 285 could be classified into three groups: 21% of these had previously diagnosed diabetes, 4% had newly diagnosed diabetes and the remaining patients were categorized as non-diabetic. Casual blood glucose >or=11.1 mmol x l(-1)at admission was found in 12 patients with no previously known diabetes, but diabetes mellitus was confirmed in only six of these patients. Haemoglobin A1c showed considerable overlapping of values between the three groups of patients (i.e. patients with known diabetes mellitus, patients with newly diagnosed diabetes mellitus and non-diabetics). CONCLUSION: One of four patients with acute myocardial infarction had diabetes mellitus. Increased casual blood glucose at admission was not a reliable measure to establish a diagnosis of diabetes and thus follow-up measures were necessary. Haemoglobin A1c was found to be an unreliable measure in the verification of diabetes. PMID- 11428851 TI - Long-term effects of in-hospital cardiac rehabilitation on the cardiac risk profile. A case-control study in pairs of siblings with myocardial infarction. AB - AIMS: In the general population, measures for secondary prevention of myocardial infarction are poorly utilized. Our aim was therefore to analyse whether post myocardial infarction in-hospital rehabilitation and education programmes improve the subsequent utilization of preventive strategies. METHODS AND RESULTS: We screened 93 500 patient charts in cardiac rehabilitation clinics to identify a myocardial infarction patient with a sibling, who likewise had a myocardial infarction prior to the age of 60 years but was discordant with respect to the participation in cardiac in-hospital rehabilitation. In 92 such sibling pairs the coronary risk profile was studied by standardized questionnaire, biochemical measurements and physical examination. At the time of the acute myocardial infarction, both groups showed an equal risk factor distribution. However, at follow-up (on average 5.5 years after myocardial infarction), rehabilitation siblings presented with significantly lower systolic (137+/-2 vs 150+/-3 mmHg, P<0.01) and diastolic blood pressure (82+/-1 vs 89+/-1 mmHg, P<0.01). Antihypertensive drug therapy resulted more often in effective ( DMRIE 50 ranging at approximately 3 mU per beta-gal per well. Low expression of beta-galactosidase was achieved using DAC 30, DOSGA 30 and DOSGA 100. No beta-galactosidase expression was found in control dishes. There was no difference seen following transfection of primary or subsequent passages of BCEC. As indicated by the acid phosphatase assay, no significant toxicity was detected for the most efficient lipids used. Of the preparations studied, SP 20 appeared as the optimal vehicle for plasmid-mediated transfection of BCEC. The ability to deliver genes to BCEC via liposomes could be valuable, since the use of other vectors for transfection may be limited by undesired effects. PMID- 11428858 TI - Isolation and characterization of a mouse monoclonal antibody against human corneal endothelial cells. AB - In vitro cultivation of human corneal endothelial cells (HCEC) is associated with loss of typical cobblestone-like appearance during successive passages. Thus far morphology was the sole criterion for the cell's endothelial nature. Mouse monoclonal antibodies (mabs) to human corneal endothelial cells were raised using standard immunization and hybridoma isolation procedures. The specificity of mabs for human corneal endothelial cells was tested in comparison to other endothelial cell types, to fibroblasts, corneal keratocytes and to human retinal pigmented epithelial cells. In addition immunofluorescence or immunoperoxidase staining was performed with frozen tissue sections of human corneas and with various other human tissues. The mab 9.3.E reacts with cultured human corneal endothelial cells, but not with cultured human fibroblasts and human keratocytes. In frozen sections selective positivity of corneal endothelium in contrast to negativity of the other corneal cell types was confirmed. In investigated extraocular tissues positivity was observed in smooth muscle cells including related cells (i.e. Ito and mesangial cells) and in Schwann's cells and adipocytes, but apparently not in vascular endothelial cells. The mab is human-specific and binds to a protein with a molecular weight of 130 kDa mainly accumulating along cell membranes. A mouse monoclonal antibody against human corneal endothelial cells was established in vitro and was shown to be capable of differentiating corneal endothelial cells from other corneal cell types, especially from corneal keratocytes. It is, however, not cornea-specific, but also reacts with certain extraocular cell types. PMID- 11428859 TI - Cell density regulated expression of transcription factor Sp1 in corneal stromal cultures. AB - Sp1, a ubiquitously expressed transcription factor, has been implicated to have a role in cell differentiation and cell proliferation. In keratoconus, a corneal disease characterized by thinning and scarring of the central cornea, Sp1 is found up-regulated. In the present study, we examined the expression of Sp1 in stromal cells cultured from normal human and keratoconus-afflicted corneas and evaluated the influence of varying cell densities. Immunohistochemical staining, Western blotting and electrophoretic mobility shift assays indicated that in both normal human and keratoconus cultures, Sp1 protein levels and binding activities increased with the density of cells. The basal level of Sp1 in keratoconus cultures was higher than that in normals. These results demonstrate a marked density mediated up-regulation of Sp1 in corneal stromal cells, suggesting that the Sp1 expression may be regulated by differentiation states of the cells in the cornea. In addition, cells from keratoconus corneas in vitro appear to carry and retain the Sp1 abnormality as in vivo. The Sp1 defect may be an inborn error in keratoconus. PMID- 11428860 TI - Activation of extracellular signal-regulated kinase in trabecular meshwork cells. AB - A number of different agents, such as growth factors, cytokines and phorbol esters have been shown to modulate trabecular meshwork cell function. These studies were designed to evaluate the role extracellular signal-regulated kinase (ERK) pathway plays in mediating the responses to platelet-derived growth factor BB (PDGF-BB) and phorbol 12-myristate 13-acetate (PMA) in trabecular meshwork cells. The human trabecular meshwork cell line, HTM-3, and the bovine trabecular meshwork (BTM) cells were treated with either PDGF-BB or PMA and the activation of ERK 1/2 evaluated. The effects of the MAP kinase kinase (MEK) inhibitor U0126, and the PKC inhibitor chelerythrine on ERK 1/2 were also determined. In a separate group of experiments, cells were treated with PDGF-BB or PMA and the secretion of matrix metalloproteinase-2 (MMP-2) evaluated. The addition of PDGF BB or PMA produced time- and dose-dependent activation of ERK 1/2. Pretreatment with U0126 or chelerythrine significantly reduced ERK 1/2 activation induced by PDGF-BB or PMA. The addition of PDGF-BB or PMA stimulated the secretion of MMP-2. This secretory response was inhibited by pretreatment with the MEK inhibitor U0126. In trabecular meshwork cells, PDGF-BB and PMA activate ERK 1/2 by a PKC dependent mechanism. Activation of ERK 1/2 by these agents in trabecular meshwork cells leads to the secretion of MMP-2. These studies provide evidence that ERK pathway is an important mechanism for integrating various signals that regulate trabecular function. PMID- 11428861 TI - The effect of high and very low fluorescent light exposure levels on age-related cataract in a pigmented mouse strain. AB - This study examined the effect of fluorescent light on the timing and severity of age-related cataracts in a fully pigmented mouse strain, the (C57BL/6 x C3H)F1, that normally develops slowly progressing age-related cataracts only beyond middle age. Two groups of 56 animals each were exposed, respectively, either to a daily range of 66-222 foot candles (FC) or to 1 FC of standard fluorescent lighting for a period beginning at 5 weeks of age and ending at 33.5 months (by which time approximately 65% of the colony had died). Contrary to previous reports involving albino rats or mice and a strain of pigmented but cataract prone transgenic mice, the two groups of animals in this experiment did not differ for cataract development in time of first occurrence, rate of advancement, or degree of severity. It was concluded that genetic predisposition, based on levels of oxidative free radical production vs antioxidant enzyme and repair enzyme protection in the lens, was probably the major factor governing the rate and degree of age-related cataract development in these animals. The effect of relatively intense life-long fluorescent light exposure was so minimal as not to be manifested in this strain of mice under the conditions of this experiment. Remarkably, maintaining the one group of mice in semi-darkness from 5 weeks of age to beyond their mean lifespans did nothing to delay or reduce the incidence or severity of their age-related cataracts. PMID- 11428862 TI - Apoptotic cell death in the mouse retinal ganglion cell layer is induced in vivo by the excitatory amino acid homocysteine. AB - Homocysteine, an excitatory amino acid and a homolog of cysteine, induces neuronal cell death in brain via stimulation of N-methyl-D-aspartate (NMDA) receptors. It also selectively activates NMDA receptors of retinal ganglion cells, but it is not known if high levels of homocysteine are toxic to these cells. The purpose of this study was to determine whether increased levels of homocysteine caused death of neurons in the ganglion cell layer; if so whether this death occurred via an apoptotic mechanism and to determine the consequences of simultaneous elevation of homocysteine and glutamate, a known retinal excitotoxin, on the viability of neurons of the ganglion cell layer. C57BL/6 mice were injected intravitreally with either homocysteine or glutamate/homocysteine combined (final concentrations: 25, 75, and 200 microM); injection of glutamate (25 and 200 microM) served as a positive control. Eyes were harvested and cryosections prepared 5-6 days post-injection. Systematic morphometric analysis of retinas of mice injected with homocysteine indicated that the total number of cells in the ganglion cell layer decreased by about 23% following exposure to 200 microM homocysteine. To determine whether the neurons of the ganglion cell layer were dying by apoptosis, the TUNEL method was used and was confirmed by immunohistochemical studies of caspase-3, known to be expressed at high levels during retinal ganglion cell apoptosis. Microscopic analysis revealed significantly more TUNEL-positive cells in the ganglion cell layer in homocysteine-injected eyes than in contralateral PBS-injected eyes. Retinas injected with 75 and 200 microM homocysteine displayed significantly more TUNEL positive neurons in the ganglion cell layer (2 and 2.9, respectively) than PBS injected retinas (0.25). In eyes injected simultaneously with homocysteine/glutamate, the number of apoptotic cells in the ganglion cell layer almost doubled that for homocysteine or glutamate injections alone. Immunohistochemical analysis of activated caspase-3 revealed numerous positively labelled neurons in the ganglion cell layer in homocysteine and homocysteine/glutamate-injected eyes, but not in PBS-injected eyes. Quantification of this data revealed a significantly greater number of caspase-3 positive neurons in the ganglion cell layer of retinas injected with 75 and 200 microM homocysteine (2.9 and 4.4, respectively) than for PBS-injected retinas (0.5). This confirms that death of neurons in the ganglion cell layer is occurring by apoptosis. The present study provides the first evidence that homocysteine is toxic to neurons of the ganglion cell layer. In addition, it provides evidence that these retinal neurons are dying by apoptosis and it demonstrates for the first time that excitotoxic damage to neurons of the ganglion cell layer is potentiated by simultaneous elevation of homocysteine and glutamate. These findings are relevant to retinal ganglion cell death characteristic of diabetic retinopathy, which is thought to be mediated by overstimulation of the NMDA receptor. PMID- 11428863 TI - [(3)H]-serotonin release from bovine iris-ciliary body: pharmacology of prejunctional serotonin (5-HT(7)) autoreceptors. AB - In the present study, we investigated the pharmacological characteristics of electrically stimulated [(3)H]-serotonin release from mammalian iris-ciliary bodies. Isolated bovine and human iris-ciliary bodies were loaded with [(3)H] serotonin, superfused with Krebs buffer solution and then stimulated with trains of 300 direct current (d.c.) pulses to initiate the release of the transmitter. The modification of this [(3)H]-serotonin release process by various serotonergic agonists and antagonists was studied in order to define the pharmacology of serotonin receptor(s) present in the iris-ciliary body. In bovine iris-ciliary body, electrically-evoked [(3)H]-serotonin release was calcium-dependent, tetrodotoxin-sensitive and was enhanced by serotonin (EC(50) = 200 n M) and 5 carboxmidotryptamine (EC(50) = 4 n M). The rank order of potency of agonists in enhancing field-stimulated [(3)H]-serotonin release was: 5-carboamidotryptamine > m-chlorophenylbiguanide > 2-methyl-5-hydroxytryptamine = 5-methoxy dimethyltryptamine > serotonin > 5-methoxy-tryptamine > L-694,247 = alpha-methyl 5-hydroxytryptamine > CGS 12066A = 8-hydroxy-2-(di- n -propylamino)tetraline. Serotonin and m-chlorophenylbiguanide also enhanced electrically-evoked [(3)H] serotonin release from human iris-ciliary bodies with EC(50)s of 3 microM and 30 n M, respectively. The pharmacological profile displayed by serotonin receptor agonists was supported by the potent antagonism of the serotonin-induced enhancement of [(3)H]-serotonin release by 5HT(7)receptor antagonists SB-258718 (IC(50) = 18.6 +/- 1.2 nM; n = 4) and mesulergine (IC(50) = 0.26 +/- 0.05 nM; n = 4). However, antagonists at 5HT(6)and 5HT(3)receptors exhibited a relatively weak blockade of serotonin induced enhancement of field-stimulated [(3)H]-serotonin release. These studies have shown the presence of functionally active prejunctional 5HT(7)autoreceptors regulating the release of [(3)H]-serotonin from bovine iris-ciliary bodies. Excitatory prejunctional 5-HT autoreceptors also exist in human iris-ciliary bodies. It is possible that these serotonin autoreceptors may have relevance to the regulation of aqueous humor dynamics in the anterior uvea. PMID- 11428864 TI - Suppressive effects of histamine H1 receptor antagonist diphenhydramine on the leukocyte infiltration during endotoxin-induced uveitis. AB - Histamine has been shown to play an important role in the step of leukocyte rolling, the initial step to leukocyte infiltration into an inflamed region. We investigated the roles of histamine in the leukocyte recruitment during endotoxin induced uveitis (EIU) in vivo using acridine orange digital fluorography. An injection of histamine into the vitreous cavity of a Lewis rat induced leukocyte rolling along the major retinal veins. In other experiments, EIU was induced in Lewis rats by footpad injection of lipopolysaccharide (LPS). Leukocyte rolling was also observed in the retinal veins of EIU rats. To block the histamine H1 receptor, diphenhydramine (DPH) was administered intraperitoneally 15 min before the LPS injection. DPH significantly inhibited leukocyte rolling along the major retinal veins of EIU rats, suppressing leukocyte infiltration into the vitreous cavity. The vasodilation in EIU was also significantly suppressed with DPH. Moreover, leukocyte infiltration into aqueous humor was significantly suppressed in DPH-treated rats. Although the inhibitory effects of DPH was less obvious at later time points, addition of DPH every 12 hr showed prolonged anti-inflammatory effects up to 48 hr after LPS injection. In contrast, protein leakage into the aqueous humor was not suppressed as much as leukocyte infiltration with DPH. These results suggest that histamine would play a pivotal role in leukocyte recruitment during EIU in rats. Blocking the histamine H1 receptor might help to prevent or minimize leukocyte infiltration in uveitis. PMID- 11428865 TI - Expression of the 67 kDa laminin receptor (67LR) during retinal development: correlations with angiogenesis. AB - Interaction of vascular cells with the laminin component of basement membranes is important for normal cell function. Likewise, abnormal interactions may have a critical role in vascular pathology. It has been previously demonstrated that the 67 kDa laminin receptor (67LR) is expressed at high levels during proliferative retinopathy in a mouse model and in the current study we have examined 67LR in the neonatal mouse to determine if this receptor plays a role in aspects of developmental angiogenesis in the developing murine retina. Groups of C57/BL6 mice were killed at postnatal day P1, P3, P5, P7, P9 and P11 to assess the retinal vasculature. A number of mice were perfused with FITC-dextran and the eyes removed, fixed in 4% paraformaldehyde (PFA) and flat-mounted for confocal scanning laser microscopy. The eyes from the remaining mice were either placed in 4% PFA and embedded in paraffin-wax, or had the neural retina dissected off and total RNA or protein extracted. Immunofluorescence, in situ hybridization, quantitative reverse transcriptase polymerase chain reaction and Western blotting analysis were employed to locate and determine expression levels of 67LR. Both 67LR mRNA and protein expression showed a characteristic bi-phasic expression pattern which correlated with key stages of retinal vascular development in the murine retina. 67LR showed high expression levels at P1 (P < 0.05) (correlating with superficial vascular plexus formation) and at P7 (P < 0.05) (correlating with deep vascular plexus formation). Conversely, 67LR expression was decreased when active angiogenic activity was lowest. Significantly, optical sectioning of retinal flat-mounts revealed high levels of 67LR expression in developing segments of both superficial and deep capillary plexi, a pattern which co localized strongly with laminin. 67LR is regulated during post-natal development of the retinal vasculature. High levels of 67LR during the two well-defined phases of retinal capillary plexus formation suggests that this receptor may play an important role in retinal angiogenesis. PMID- 11428866 TI - A functional study on prostanoid receptors involved in cultured human iridal melanocyte stimulation. AB - The effects of various prostanoids on the growth, melanogenesis and dendrification of cultured iridal melanocytes were studied. Iridal melanocytes were isolated and cultured with medium supplemented with cAMP elevating agents and basic fibroblast growth factor (bFGF) (complete medium). The iridal melanocytes were plated into multiple well plates and cultured with complete medium or various deleted media with or without various prostanoids at different concentrations. After 6 days, the numbers of cells and dendrites were counted and melanin content was measured and compared with controls. Prostaglandin E(2), an EP(2)receptor agonist (AH 13205) and AGN 192093 (thromboxane mimetic) stimulated growth, melanogenesis and dendrification of cultured iridal melanocytes in cAMP deleted medium. A mixed EP(1)and EP(3)receptor agonist (sulprostone), a EP(4)receptor agonist (ONO-AE1-329), IP receptor agonists (cicaprost or iloprost) and a TP receptor agonist (U-46619) showed no effect. Prostaglandin D(2)showed stimulating effects. However, these stimulating effects could not be blocked by the addition of a DP receptor antagonist (BW A868C). Furthermore, a DP receptor agonist (BW 245C) showed no effects, indicating that the effect of prostaglandin D(2)may involve receptors other than the DP receptor subtype. The present study indicates that: (1) among various EP receptor agonists, only an EP(2)receptor agonist has stimulating effects on iridal melanocytes; (2) DP, IP and TP receptor agonists do not have stimulating effects; and (3) the mechanisms of action of prostaglandin D(2)and AGN 192093 need further study. PMID- 11428867 TI - Macrophage depletion prevents leukocyte adhesion and disease induction in experimental melanin-protein induced uveitis. AB - The purpose was to study the effects of macrophage depletion with liposomal dichloromethylene-diphosphonate (Cl(2)MDP-lip) on inflammation and leukocyte endothelium interaction in experimental melanin-protein induced uveitis (EMIU). Lewis rats (n = 48) were immunized with melanin-associated protein in complete Freund's adjuvant and pertussis toxin. Control groups received adjuvants without the antigen (n = 12) or no injection (n = 6). Animals received treatment with either CL(2)MDP-lip or empty liposomes (empty-lip) on day -2, 1, 4, 6 and 8. Leukocytes were stained with rhodamine 6G i.v. and intravital fluorescence microscopy (IVM) was performed on day 4, 6, 8 and 10 to quantify leukocyte rolling and arrest. After IVM, the cell count and protein concentration were determined in aqueous humor and plasma levels of TNF-alpha and IFN-gamma were measured by ELISA. In EMIU, leukocyte rolling increased on day 4 (10.0 +/- 1.2 cells min(-1)vs baseline of 5.7 +/- 0.7 cells min(-1), mean +/- S.E.(M.)) and peaked on day 8 (40.8 +/- 4.2 cells min(-1);P < or = 0.05). Leukocyte arrest was increased on day 8 (175.4 +/- 18.2 cells mm(-2)vs baseline of 59.7 +/- 7.1 cells mm(-2);P < or = 0.05) and day 10 (371.7 +/- 30.7 cells mm(-2)). CL(2)MDP-lip prevented leukocyte rolling (day 10: 16.6 +/- 1.8 cells min(-1)vs 30.7 +/- 2.9 cells min(-1); CL(2)MDP-lip vs untreated EMIU;P < or = 0.05) and arrest (day 8: 88.3 +/- 13 cells mm(-2); day 10: 128.5 +/- 12.9 cells mm(-2);P < or = 0.05). Empty-lip had no effect on leukocyte rolling (day 10: 34.8 +/- 4.2 cells min(-1)) or arrest (day 8: 159.3 +/- 12.9 cells mm(-2), day 10: 421.2 +/- 41.6 cells mm( 2)). CL(2)MDP-lip completely suppressed leukocyte emigration (11 +/- 2 cells microl(-1)vs 100 +/- 29 cells microl(-1); CL(2)MDP-lip vs empty-lip;P < or = 0.05) and protein extravasation into aqueous humor (2.7 +/- 0.3 mg ml(-1)vs 14.2 +/- 2.1 mg ml(-1); CL(2)MDP-lip vs empty-lip;P < or = 0.05), abrogated the TNF alpha response (32.5 +/- 2.7 pg ml(-1)vs 954.9 +/- 216.3 pg ml(-1); CL(2)MDP-lip vs untreated EMIU;P < or = 0.05) and caused an attenuated and delayed elevation of IFN-gamma. CL(2)MDP-lip prevented the inflammatory reaction of EMIU and inhibited the increase of leukocyte-endothelium interaction in iris vessels. Our findings emphasize the pivotal role macrophages play in the initiation of autoimmune disease. PMID- 11428868 TI - Activation of p38, ERK1/2 and NIK pathways is required for IL-1beta and TNF-alpha induced chemokine expression in human retinal pigment epithelial cells. AB - Chemokine secretion by human retinal pigment epithelium (hRPE) in response to IL 1beta and TNF-alpha occurs in infectious and noninfectious retinal diseases. In this study, the roles of p38 kinase and extracellular signal-regulated kinase (ERK) signaling pathways were investigated for IL-1beta- or TNF-alpha-induced IL 8 and MCP-1 secretion by hRPE cells. Treatment of hRPE cells with IL-1beta or TNF alpha caused increased steady-state IL-8 and MCP-1 mRNA levels and protein secretion. Stimulation of hRPE with IL-1beta and TNF-alpha resulted in degradation of IkappaB-alpha, nuclear translocation of NF-kappaB, and prominent increases in p38 and ERK1/2 phosphorylation for as little as 3 min. The induced IL-8 and MCP-1 mRNA and proteins were partially suppressed by U0126, a specific MEK inhibitor, and by SB202190, a selective p38 inhibitor. This induction was completely blocked by simultaneous administration of the two drugs or by incubation with inhibitors for activation of NF-kappaB such as BAY11-7085, CAPE, and parthenolide. These results suggest that co-activation of MEK/ERK and p38 pathways as well as activation of NIK pathway are essential for IL-1beta- and TNF alpha-stimulation of IL-8 and MCP-1 gene expression in hRPE cells. Furthermore, co-administration of U0126 and SB202190 did not affect the induced degradation of IkappaB-alpha and NF-kappaB nuclear translocation, indicating that NF-kappaB is activated by IL-1beta and TNF-alpha independently of activation of MEK/MAPK and p38 pathways in hRPE cells. PMID- 11428869 TI - 18 kDa protein tyrosine phosphatase in the ocular lens. AB - Protein tyrosyl phosphorylation and dephosphorylation play essential roles in regulating cellular events such as proliferation and differentiation, and their involvement in the lens development and transparency is also suggested. The level of tyrosine phosphorylation in a given protein is regulated by the opposing actions of protein-tyrosine kinases (Tyr kinases) and protein-tyrosine phosphatases (TyrPases). Recent studies have revealed that some Tyr kinases, such as platelet-derived growth factor receptor and fibroblast growth factor receptor, are present in the lens, however, little is known about TyrPases in the lens. In this study, we found a 18 kDa protein tyrosine phosphatase (18 kDa TyrPase) predominantly present in the ocular lens of various animals. We purified the phosphatase from the lens of chick embryo and characterized its activity.Phosphatase activity was determined in chick embryo, mouse, rabbit and bovine lenses using p -nitrophenyl phosphate (p NPP) as substrate. All lenses examined dephosphorylated p NPP under acidic conditions, and a large portion of the activity resided in a low molecular weight protein, ca. 18 kDa, following high-resolution gel permeation column chromatography. The brain and liver showed high dephosphorylation activities, but most of their activity was present in high molecular weight fractions, unlike that in the lens. The 18 kDa phosphatase was purified from the lens of 17 day old chick embryos to near-homogeneity with two step rapid chromatography. This phosphatase showed strict substrate specificity for phosphotyrosine and phosphotyrosyl peptides, suggesting that it was a kind of protein tyrosine phosphatases (TyrPases). Several known inhibitors of TyrPases, such as SH blockers, vanadate and phenylarsine oxide, strongly inhibited the enzyme activity. The molecular weight, substrate specificity, and responses to various inhibitors and activators coincide well with those reported for the low molecular weight protein tyrosine phosphatase (LMW-TyrPase), belonging to the TyrPase superfamily. These results suggest that the 18 kDa phosphatase found in the lens is a LMW-TyrPase. The 18 kDa TyrPase is the predominant phosphatase in the ocular lens. It may be involved in regulation of lens cell proliferation, differentiation and/or lens transparency. PMID- 11428870 TI - Syneretic response to pressure in bovine and rhesus monkey vitreous. AB - Excised bovine and rhesus monkey vitreous samples were subjected to different hydrostatic pressures (0.03, 1 and 2 atm) for 12 hr. At the end of the experiment the samples were frozen in a -70 degrees C dry ice-acetone bath and thereafter maintained in frozen conditions. Differential scanning calorimetry (DSC) was used to obtain freezable water (FW) content. The total water content of the samples was obtained by thermogravimetric (TGA) techniques. The non-freezable water (NFW) content was obtained as a difference between total and freezable water contents. The total water content did not show a trend in pressure dependence, neither in bovine nor in rhesus monkey vitreous samples. However, the free water/bound water ratio, i.e. FW/NFW, increased with increasing pressure in vitreous samples of both species. The trend of the average FW/NFW values as a function of hydrostatic pressure implies a simple syneretic response, i.e. the conversion of bound (non freezable) water to free water as pressure was increased. PMID- 11428871 TI - Local fundus response to blue (LED and laser) and infrared (LED and laser) sources. AB - Light damage research began during the early years of laser light exploration. There is a clear and significant literature that identifies an easily demonstrated retina-pigment epithelium pathology which is associated with short wavelength exposures below 520 nm. Recent interest has expanded because of the growing evidence for a blue light contribution to the retina aging process by way of a poorly understood chemical process(es) that involve circulation, oxidative reactions and the spectral absorption properties of the pigment epithelium. New powerful sources of relatively inexpensive blue energy have become available as a family of light emitting diodes. In this experiment, we examined funduscopic, angiographic and scanning laser tomographic measures of the retinal-pigment epithelium response to LED and laser spectral blue and infrared emissions closely matched in wavelengths and delivered under carefully matched circumstances. Ten retinas in normal young rhesus monkeys were locally exposed to various energy density values at 458 nm (Argon laser) ranging from 5 to 54 J cm(-2). Eight rhesus eyes were exposed to LED irradiation with a peak wavelength of 460 nm ranging from 9 to 62 J cm(-2). Similarly, a matched infrared (IR) laser and IR LED pair were used to expose an additional ten eyes for comparison of the long wavelengths. IR irradiance ranged from 21 to 306 J cm(-2). There was no response to IR exposure in any of the eyes. Blue light exposure results were measured from the color fundus photographs, scanning laser tomographs and early- and late-phase fluorescein angiogram responses at 2 and 30 days after the exposure. Results scores were accumulated for the four measures at the two time periods. The resulting lesion scores when plotted against the exposure in J cm(-2)showed no demonstrable effect at irradiance lower than 10 J cm(-2)and near 100% effectiveness for irradiance greater than 30 J cm(-2). The most sensitive and enduring indicator of change was the late fluorescein angiograms. Nonparametric statistical analysis of the scores from the two samples support the conclusion that there is no difference in the consequences of LED and laser light exposures under these matched conditions. PMID- 11428872 TI - The organization of infection control in Italy. AB - This paper describes the organization of infection control in Italy with respect to regulatory requirements, the tasks and training of the infection control physician and nurse, and the function and responsibilities of the infection control committee. Moreover, the paper reports on incidence and prevalence of hospital-acquired infections (HAI), antibiotic usage and antimicrobial resistance in Italy. PMID- 11428873 TI - Molecular analysis of relapse vs re-infection in HIV-positive patients suffering from recurrent Clostridium difficile associated diarrhoea. AB - Recurrence is a major complication of Clostridium difficile associated diarrhoea, especially in human immunodeficiency virus (HIV) positive patients, and it is important to distinguish between relapse and re-infection in recurrent episodes. The aim of our study was to analyse C. difficile isolates obtained from HIV positive patients with recurrent diarrhoea in order to distinguish between relapse and re-infection. This analysis was based on the study of DNA similarities among isolates obtained from different episodes within each patient. Relapses occurred in 64% of patients, 32% suffered re-infections and a combination of relapse plus re-infection was seen in 4%. DNA typing methods can be useful tools to characterize recurrent episodes of C. difficile associated disease. PMID- 11428874 TI - Antibiotic prophylaxis and the risk of Clostridium difficile-associated diarrhoea. AB - To test the hypothesis that extended antibiotic prophylaxis increases the risk of Clostridium difficile -associated diarrhoea (CDAD), we conducted a retrospective cohort study of 2641 patients under-going cardiovascular surgery. Main outcome measures were the duration of prophylaxis (< 48 h vs. > 48 h) and the occurrence of CDAD. CDAD occurred in 31 patients (1.2%), who were significantly older (70 +/ 9 y vs. 66 +/- 10 y; P = 0.03), received more therapeutic antibiotics (2.2 +/- 1.9 vs. 0.4 +/- 0.9; P = 0.001) and had a longer postoperative hospital stay (26 +/- 19 d vs. 9 +/- 8 d; P < 0.001) than non-cases. After adjusting for confounding, we did not observe an association between prolonged prophylaxis and CDAD [adjusted odds ratio (AOR), 0.8; CI, 0.4-1.8]. In contrast, three independent predictors were identified: increasing length of hospital stay (AOR per one-day-increment, 1.03; CI, 1.01-1.05), and treatment with third generation cephalosporins (AOR, 5.9; CI, 2.2-16.0) or beta-lactam-beta-lactamase inhibitor combinations (AOR, 4.6; CI, 1.7-12.3). Our results did not confirm that extended prophylaxis after clean surgery increases the risk of CDAD, which remains an uncommon postoperative complication, associated even with short antibiotic exposure. PMID- 11428875 TI - MRSA in the critically ill. AB - MRSA colonization and infection rates were prospectively examined over an 18 month period in a general Intensive Care Unit. Of 642 admissions, 305 were in ICU for longer than 48 h and were hence included and a further three patients were already colonized at admission but stayed less than 48 h. Ninety-seven patients were colonized with MRSA including 19 who were already colonized at admission. There were 56 episodes of clinical infection in 43 patients. The mortality rates in the colonized and infected groups, were 14.8% and 16.2% respectively, while the rate in those not colonized was 23%. These figures were not statistically different. Those colonized or infected with MRSA had significantly longer ICU stays than those not colonized. Sputum colonization and infection was a major site for MRSA. There was diagnostic certainty of MRSA infection in 40% of cases emphasizing the difficulty in diagnosis of infection due to MRSA in the critically ill. Both colonization and infection with MRSA are associated with longer ICU stay but do not appear to influence mortality. PMID- 11428876 TI - Methods for identifying methicillin resistancein Staphylococcus aureus. AB - Five methods for the detection of methicillin-resistant Staphylococcus aureus were used to examine a collection of 100 clinical isolates comprising both susceptible and resistant strains. The disc diffusion test with oxacillin had a sensitivity of 93.3% and a specificity of 92.0% whereas mannitol salt agar containing oxacillin had a sensitivity of 100% and specificity 80.6% with high inoculum. With a low inoculum the sensitivity was 90.7% and specificity 96.0%. The MRSA screen test (Denka Seiken Co. Ltd., Japan) and Evigene MRSA Detection Kit (State Serum Institute, Denmark) tests were in complete agreement with results obtained with polymerase chain reaction assays amplifying mecA and nuc gene sequences. PMID- 11428877 TI - Risk factors for central vascular catheter-associated bloodstream infections among patients in a neonatal intensive care unit. AB - The aim of this study was to identify risk factors for catheter-associated bloodstream infection (CABSI) in neonates. We undertook a prospective investigation of the potential risk factors for CABSI (patient-related, treatment related and catheter-related) in a neonatal intensive care unit (NICU) using univariate and multivariate techniques. We also investigated the relationship between catheter hub and catheter exit site colonization with CABSI.Thirty-five episodes of CABSI occurred in 862 central catheters over a period of 8028 catheter-days, with a cumulative incidence of 4.1/100 catheters and an incidence density of 4.4/1000 catheter days. Factors independently associated with CABSI were: catheter hub colonization (odds ratio [OR] = 44.1, 95% confidence interval [CI] = 14.5 to 134.4), exit site colonization (OR = 14.4, CI = 4.8 to 42.6), extremely low weight (< 1000 g) at time of catheter insertion (OR = 5.13, CI = 2.1 to 12.5), duration of parenteral nutrition (OR=1.04, CI=1.0 to 1.08) and catheter insertion after first week of life (OR = 2.7, CI = 1.1 to 6.7). In 15 (43%) out of the 35 CABSI episodes the catheter hub was colonized, in nine (26%) cases the catheter exit site was colonized and in three (9%) cases colonization was found at both sites. This prospective cohort study on CABSI in a NICU identified five risk factors of which two can be used for risk-stratified incidence density description (birthweight and time of catheter insertion). It also emphasized the importance of catheter exit site, hub colonization and exposure to parenteral nutrition in the pathogenesis of CABSI. PMID- 11428878 TI - A comparison of prophylactic antibiotic regimens against airborne orthopaedic wound contamination. AB - Deep infection remains a major complication of joint replacement surgery despite advances in theatre design, surgical technique and antibiotic prophylaxis. Major randomized controlled trials to determine the most effective antibiotic prophylaxis are difficult to construct and interpret. In a conventional theatre, most orthopaedic intra-operative wound contamination arrives by the airborne route. This paper describes a unique method used to compare antibiotics against airborne bacteria. Seven antibiotics were incorporated into blood agar at concentrations equivalent to serum levels. Plates were then exposed to airborne theatre bacteria using a multiple synchronous collection technique. After incubation, the percentage kill was calculated for each antibiotic. At concentrations equivalent to serum level 1h post i.v. dose, all the antibiotics proved highly effective, with kill rates > 95%. Imipenem and co-amoxiclav significantly outperformed the other antibiotics with kill rates of 99.6% and 99.4%, respectively. At trough levels, the antibiotics achieved kill rates from 61% to 97.6%. Future randomized controlled trials comparing large numbers of antibiotics in the setting of an already low infection rate are inappropriate. This technique for comparing antibiotic prophylaxis in quick, inexpensive and repeatable. The superiority of imipenem is not unexpected, but of more interest is the effectiveness of co-amoxiclav over the presently favoured cefuroxime. PMID- 11428879 TI - Decrease in Candida albicans strains with reduced susceptibility to fluconazole following changes in prescribing policies. AB - This study was undertaken to identify prescribing policies likely to favour or limit fluconazole resistance within a clinical department. Fluconazole exposure within the infectious diseases and clinical haematology units was investigated, and data were compared with in vitro susceptibility of Candida albicans isolates obtained in these units. Fluconazole utilization was determined by the number of fluconazole treatment-days per 100 hospitalization days (penetration index). In the infectious diseases unit, separate evaluations for low-dose fluconazole (50 mg) prescribed as intermittent or prolonged treatment, and for higher-dosing schedules (fluconazole 200 mg) were made. Susceptibility of C. albicans isolates was surveyed in a broth microdilution assay by measuring the inhibitory concentration 50% (IC50). The penetration index (PI) for fluconazole 50mg declined from 1992 to 1977 in infectious diseases (P= 0.0048). In the meantime, total usage of fluconazole increased, due to increased prescribing of fluconazole 200 mg (P = 0.0724). The IC50 of C. albicans isolates tested in infectious diseases decreased between 1994 and 1996 from 7.33 mg/ml to 1.64 mg/ml (P = 0.0075). In clinical haematology, declines in C. albicans IC50 and fluconazole PI were not significant (P = 0.35 and P = 0.07, respectively). These data suggest that prolonged or repeated exposure to low-dose fluconazole, rather than total cumulative use, was associated with fluconazole resistance in the infectious diseases unit. Moreover, restoration of a normal ecology was observed when low dose prolonged or intermittent prescriptions were reduced. PMID- 11428880 TI - Rapid characterization of the genomovars of the Burkholderia cepacia complex by PCR-single-stranded conformational polymorphism (PCR-SSCP) analysis. AB - Four polymerase chain reaction (PCR) primer pairs (A-D), specific for the Burkholderia cepacia complex of organisms were designed to encompass the entire gene (approximately 1300 bp) from sequence alignments of the rec A operon of B. cepacia. Genomic bacterial DNA from type strains and wild-type B. cepacia complex isolates of previously determined genomovar status was amplified employing these four primer pairs, as well as a fifth primer set (E) already published. Primer sets B, C and E were successful in obtaining a PCR amplicon of correct estimated size of 598, 1107 and 1043 bp, respectively. Subsequent single-stranded conformational polymorphism (SSCP) analysis of PCR amplicons demonstrated unique profiles for the five genomovar types for primer pairs B and E, but was unable to differentiate distinguishable profile types for primer pair C. SSCP analysis was demonstrated to be simple, rapid and cost effective and may prove a useful method of genomovar typing of B. cepacia complex organisms from cystic fibrosis patients in busy diagnostic laboratories. PMID- 11428881 TI - Evaluation of the microbicidal efficacy of Steris System I for digestive endoscopes using GERMANDE and ASTM validation protocols. AB - In the light of more and more invasive procedures being carried out in digestive endoscopy using sterile devices, it appears necessary to put in place a process of endoscope reprocessing capable of ensuring the complete elimination of micro organisms contaminating the device. We undertook a study of the microbial efficacy of STERIS SYSTEM 1 (SS1) which purports to achieve this objective. The channels of a gastroscope and a colonoscope were contaminated with suspensions of Pseudomonas aeruginosa, Aspergillus niger and Bacillus subtilis spores. Two procedures were then followed: (1) manual washing only, and (2) treatment in SS1 without prewashing. Recoveries of organisms were made from each channel according to a standard methodology to discover any survivors. Contamination controls we re assessed to measure the logarithmic reduction between the initial contamination and that recovered from the channels. Six cycles per micro-organisms, per type of endoscope, and per type of procedure were carried out. From an initial contamination leve l of 10(6) micro-organisms per endoscope, no micro-organisms were recovered in 35 of the 36 cycles with the SS1. In one cycle with the colonoscope, three B. subtilis organisms were recovered from the channels. Washing only gave microbial reductions which varied according to the micro organism tested. The maximum reduction with washing alone was by a factor of 10(3 +/- 0.1)for B. subtilis and the minimum reduction factor was 10(3 +/- 0.3)for P. aeruginosa. Considering the results obtained with SS1 without prewashing and the efficacy obtained from washing only, the washing step offers an additional antimicrobial assurance reduction factor of between 10 and 103.1. This study shows that SS1, integrated into an overall reprocessing procedure for digestive endoscopes, is capable of delivering the complete elimination of contaminating micro-organisms in a reduced time and eliminates the toxic risk of reprocessing associated with aldehyde based disinfectants. PMID- 11428882 TI - Bacillus sphaericus bacteraemia in children with cancer: case reports and literature review. AB - In the period 1989-1999, Bacillus sphaericus was demonstrated to cause 12 out of 469 (2%) episodes of bacteraemia in children with cancer or receiving bone marrow transplant at G. Gaslini Children's Hospital, Genoa, Italy. Neutropenia was present in five episodes, six episodes, (all without neutropenia) were related to the presence of a central venous catheter, and one episode occurred in a patient with intestinal graft vs. host disease and gut colonization. All patients survived. Ciprofloxacin was the only drug active against all the isolated strains.Bacillus sphaericus represents a new cause of infection in the immunocompromised host, with low aggressiveness, but a peculiar pattern of antibiotic susceptibility. PMID- 11428883 TI - Diploma in Hospital Infection Control: a progress report. AB - The Diploma in Hospital Infection Control (DipHIC) was established by the Hospital Infection Society (HIS), the London School of Hygiene and Tropical Medicine (LSHTM) and the Public Health Laboratory Service (PHLS) in 1997 and has now completed two examinations. We outline progress since the announcement of the diploma and changes to the written examination and reflective portfolio. The reflective process is described and guidance provided to active infection control practitioners wishing to consider application for the diploma by accreditation of prior learning. PMID- 11428884 TI - Myobacterium chelonae and Acremonium species isolated from endoscope autodisinfector rinse water despite daily treatment with chlorine dioxide. PMID- 11428885 TI - Sterilox disinfection of endoscopes. PMID- 11428886 TI - An outbreak of multiply resistant Enterobacter cloacae in a neonatal intensive care unit. PMID- 11428889 TI - Rational design and molecular characterization of a chimaeric response regulator protein. AB - BvgA and EvgA are closely related response regulators from Bordetella pertussis and Escherichia coli. To analyze the domain borders and linker sequences of these proteins, we used limited proteolysis and matrix-assisted laser desorption/ionization-mass spectrometry analysis of the in-gel-digested proteolytic fragments. The thermolysin-sensitive linker regions were found to extend from Leu130 to Thr144 for BvgA and from Leu127 to Ser133 for EvgA. These data provided the rationale for the construction of the chimaeric protein HA. HA carries the EvgA receiver and BvgA output domains, fused in the central part of the linker sequences of the parent proteins. Thermolysin-sensitive sites of HA were found at positions identical with those in the EvgA and BvgA linker sequences, indicating intact folding of its receiver and output domains. Consistent with this, the chimaera showed virtually unchanged phosphorylation and dimerization properties. However, BvgA and HA differed in the effect of phosphorylation on their DNA-binding activities. In the case of BvgA, phosphorylation resulted in an increased affinity and specificity in DNA binding, whereas the DNA-binding properties of HA were not affected by phosphorylation. The chimaera HA was unable to activate transcription of the BvgA-dependent fha promoter, either in vivo or in vitro. These results indicate that the phosphorylation-induced activation of BvgA requires specific interactions between the receiver and output domains that are disturbed in the chimaera. PMID- 11428890 TI - Modular structure, local flexibility and cold-activity of a novel chitobiase from a psychrophilic Antarctic bacterium. AB - The gene archb encoding for the cell-bound chitobiase from the Antarctic Gram positive bacterium Arthrobacter sp. TAD20 was cloned and expressed in Escherichia coli in a soluble form. The mature chitobiase ArChb possesses four functionally independent domains: a catalytic domain stabilized by Ca(2+), a galactose-binding domain and an immunoglobulin-like domain followed by a cell-wall anchorage signal, typical of cell-surface proteins from Gram-positive bacteria. Binding of saccharides was analyzed by differential scanning calorimetry, allowing to distinguish unequivocally the catalytic domain from the galactose-binding domain and to study binding specificities. The results suggest that ArChb could play a role in bacterium attachment to natural hosts. Kinetic parameters of ArChb demonstrate perfect adaptation to catalysis at low temperatures, as shown by a low activation energy associated with unusually low K(m) and high k(cat) values. Thermodependence of these parameters indicates that discrete amino acid substitutions in the catalytic center have optimized the thermodynamic properties of weak interactions involved in substrate binding at low temperatures. Microcalorimetry also reveals that heat-lability, a general trait of psychrophilic enzymes, only affects the active site domain of ArChb. PMID- 11428891 TI - Effect of mutations in the Mu-host junction region on transpososome assembly. AB - Mu transposition occurs through a series of higher-order nucleoprotein complexes called transpososomes. The region where the Mu DNA joins the host DNA plays an integral role in the assembly of these transpososomes. We have created a series of point mutations at the Mu-host junction and characterized their effect on the Mu in vitro strand transfer reaction. Analysis of these mutant constructs revealed an inhibition in transpososome assembly at the point in the reaction pathway when the junction region is engaged by the transposase active site (i.e. the transition from LER to type 0). We found that the degree of inhibition was dependent upon the particular base-pair change at each position and whether the substitution occurred at the left or right transposon end. The MuB transposition protein, an allosteric effector of MuA, was shown to suppress all of the inhibitory Mu-host junction mutants. Most of the mutant constructs were also suppressed, to varying degrees, by the substitution of Mg(2+) with Mn(2+). Analysis of the mutant constructs has revealed hierarchical nucleotide preferences at positions -1 through +3 for transpososome assembly and suggests the possibility that specific metal ion-DNA base interactions are involved in DNA recognition and transpososome assembly. PMID- 11428892 TI - Domain combinations in archaeal, eubacterial and eukaryotic proteomes. AB - There is a limited repertoire of domain families that are duplicated and combined in different ways to form the set of proteins in a genome. Proteins are gene products, and at the level of genes, duplication, recombination, fusion and fission are the processes that produce new genes. We attempt to gain an overview of these processes by studying the evolutionary units in proteins, domains, in the protein sequences of 40 genomes. The domain and superfamily definitions in the Structural Classification of Proteins Database are used, so that we can view all pairs of adjacent domains in genome sequences in terms of their superfamily combinations. We find 783 out of the 859 superfamilies in SCOP in these genomes, and the 783 families occur in 1307 pairwise combinations. Most families are observed in combination with one or two other families, while a few families are very versatile in their combinatorial behaviour; 209 families do not make combinations with other families. This type of pattern can be described as a scale-free network. We also study the N to C-terminal orientation of domain pairs and domain repeats. The phylogenetic distribution of domain combinations is surveyed, to establish the extent of common and kingdom-specific combinations. Of the kingdom-specific combinations, significantly more combinations consist of families present in all three kingdoms than of families present in one or two kingdoms. Hence, we are led to conclude that recombination between common families, as compared to the invention of new families and recombination among these, has also been a major contribution to the evolution of kingdom-specific and species-specific functions in organisms in all three kingdoms. Finally, we compare the set of the domain combinations in the genomes to those in the RCSB Protein Data Bank, and discuss the implications for structural genomics. PMID- 11428893 TI - E. coli Rep oligomers are required to initiate DNA unwinding in vitro. AB - E. coli Rep protein is a 3' to 5' SF1 superfamily DNA helicase which is monomeric in the absence of DNA, but can dimerize upon binding either single-stranded or duplex DNA. A variety of biochemical studies have led to proposals that Rep dimerization is important for its helicase activity; however, recent structural studies of Bacillus stearothermophilus PcrA have led to suggestions that SF1 helicases, such as E. coli Rep and E. coli UvrD, function as monomeric helicases. We have examined the question of whether Rep oligomerization is important for its DNA helicase activity using pre-steady state stopped-flow and chemical quenched flow kinetic studies of Rep-catalyzed DNA unwinding. The results from four independent experiments demonstrate that Rep oligomerization is required for initiation of DNA helicase activity in vitro. No DNA unwinding is observed when only a Rep monomer is bound to the DNA substrate, even when fluorescent DNA substrates are used that can detect partial unwinding of the first few base-pairs at the ss-ds-DNA junction. In fact, under these conditions, ATP hydrolysis causes dissociation of the Rep monomer from the DNA, rather than DNA unwinding. These studies demonstrate that wild-type Rep monomers are unable to initiate DNA unwinding in vitro, and that oligomerization is required. PMID- 11428894 TI - Micromechanics and ultrastructure of actin filament networks crosslinked by human fascin: a comparison with alpha-actinin. AB - Fascin is an actin crosslinking protein that organizes actin filaments into tightly packed bundles believed to mediate the formation of cellular protrusions and to provide mechanical support to stress fibers. Using quantitative rheological methods, we studied the evolution of the mechanical behavior of filamentous actin (F-actin) networks assembled in the presence of human fascin. The mechanical properties of F-actin/fascin networks were directly compared with those formed by alpha-actinin, a prototypical actin filament crosslinking/bundling protein. Gelation of F-actin networks in the presence of fascin (fascin to actin molar ratio >1:50) exhibits a non-monotonic behavior characterized by a burst of elasticity followed by a slow decline over time. Moreover, the rate of gelation shows a non-monotonic dependence on fascin concentration. In contrast, alpha-actinin increased the F-actin network elasticity and the rate of gelation monotonically. Time-resolved multiple-angle light scattering and confocal and electron microscopies suggest that this unique behavior is due to competition between fascin-mediated crosslinking and side branching of actin filaments and bundles, on the one hand, and delayed actin assembly and enhanced network micro-heterogeneity, on the other hand. The behavior of F-actin/fascin solutions under oscillatory shear of different frequencies, which mimics the cell's response to forces applied at different rates, supports a key role for fascin-mediated F-actin side-branching. F-actin side-branching promotes the formation of interconnected networks, which completely inhibits the motion of actin filaments and bundles. Our results therefore show that despite sharing seemingly similar F-actin crosslinking/bundling activity, alpha-actinin and fascin display completely different mechanical behavior. When viewed in the context of recent microrheological measurements in living cells, these results provide the basis for understanding the synergy between multiple crosslinking proteins, and in particular the complementary mechanical roles of fascin and alpha-actinin in vivo. PMID- 11428895 TI - DNA G-quartets in a 1.86 A resolution structure of an Oxytricha nova telomeric protein-DNA complex. AB - The Oxytricha nova telomere end binding protein (OnTEBP) recognizes, binds and protects the single-stranded 3'-terminal DNA extension found at the ends of macronuclear chromosomes. The structure of this complex shows that the single strand GGGGTTTTGGGG DNA binds in a deep cleft between the two protein subunits of OnTEBP, adopting a non-helical and irregular conformation. In extending the resolution limit of this structure to 1.86 A, we were surprised to find a G quartet linked dimer of the GGGGTTTTGGGG DNA also packing within the crystal lattice and interacting with the telomere end binding protein. The G-quartet DNA exhibits the same structure and topology as previously observed in solution by NMR with diagonally crossing d(TTTT) loops at either end of the four-stranded helix. Additionally, the crystal structure reveals clearly visible Na(+), and specific patterns of bound water molecules in the four non-equivalent grooves. Although the G-quartet:protein contact surfaces are modest and might simply represent crystal packing interactions, it is interesting to speculate that the two types of telomeric DNA-protein interactions observed here might both be important in telomere biology. PMID- 11428896 TI - Specific and non-specific interactions of integration host factor with DNA: thermodynamic evidence for disruption of multiple IHF surface salt-bridges coupled to DNA binding. AB - Site-specific DNA binding of architectural protein integration host factor (IHF) is involved in formation of functional multiprotein-DNA assemblies in Escherichia coli, while non-specific binding of IHF and other histone-like proteins serves to structure the nucleoid. Here, we report an isothermal titration calorimetry study of the thermodynamics of binding IHF to a 34 bp fragment composed entirely of the specific H' site from lambda-phage DNA. At low to moderate [K(+)] (60-100 mM), strong competition is observed between specific and non-specific binding as a result of a low specificity ratio (approximately 10(2)) and a very small non specific site size. In this [K(+)] range, both specific and non-specific binding are enthalpy-driven, with large negative enthalpy, entropy and heat capacity changes and binding constants that are insensitive to [K(+)]. Above 100 mM K(+), only specific binding is observed, and both the binding constant and the magnitudes of enthalpy, entropy and heat capacity changes all decrease strongly with increasing [K(+)]. When interpreted in the context of the structure of the specific complex, the thermodynamics provide compelling evidence for a previously unrecognized design principle by which proteins that form extensive binding interfaces with nucleic acids control binding constants, binding site sizes and effects of temperature and ion concentrations on stability and specificity. We propose that up to 22 of the 23 IHF cationic side-chains that are located within 6 A of DNA phosphate oxygen atoms in the complex, are masked in the absence of DNA by pairing with anionic carboxylate groups in intramolecular salt-bridges (dehydrated ion-pairs). These salt-bridges increase in stability with increasing temperature and decreasing [K(+)]. To explain the unusual thermodynamics of IHF DNA interactions, we propose that both specific and non-specific binding at low [K(+)] require disruption of salt-bridges (as many as 18 for specific binding) whereupon many of the unmasked charged groups hydrate and the cationic groups interact with DNA. From structural or thermodynamic parallels with IHF, we propose that large-scale coupling of disruption of protein salt-bridges to DNA binding is significant for other large-interface DNA wrapping proteins including the nucleosome, lac repressor core tetramer, RNA polymerase core protein, HU and SSB. PMID- 11428897 TI - Plasmid transcriptional repressor CopG oligomerises to render helical superstructures unbound and in complexes with oligonucleotides. AB - CopG is a 45 amino acid residue transcriptional repressor involved in the copy number control of the streptococcal plasmid pMV158. To do so, it binds to a DNA operator that contains a 13 bp pseudosymmetric DNA element. Binding of CopG to its operator results in repression, at the transcriptional level, of its own synthesis and that of the initiator of replication protein, RepB. Biochemical experiments have shown that CopG co-operatively associates to its target DNA at low protein:DNA ratios, completely protecting four helical turns on the same face of the double helix in both directions from the inverted repeat that constitutes the CopG primary target. This has been correlated with a CopG-mediated DNA bend of about 100 degrees. Here, we show that binding of CopG to DNA fragments containing the inverted repeat just at one end led to nucleation of the protein initiating from the inverted repeat. Nucleation extended to the entire fragment, with CopG-DNA contacts occurring on the same face of the DNA helix. The protein, the prototype for a family of homologous plasmid repressors, displays a homodimeric ribbon-helix-helix arrangement. It polymerises within the unbound crystal to render a continuous right-handed protein superhelix of homodimers, around which a bound double-stranded (ds) DNA could wrap. We have solved the crystal structure of CopG in complex with a 22 bp dsDNA oligonucleotide encompassing the cognate pseudosymmetric element. In the crystal, one protein tetramer binds at one face of the DNA with two parallel beta-ribbons inserted into the major groove. The DNA is bent about 50 degrees under compression of both major and minor grooves. A continuous right-handed complex helix made up mainly by protein-protein and some protein-DNA interactions is observed. The protein protein interactions involve regions similar to those observed in the oligomerisation of the native crystals and those employed to set up the functional tetramer. A previously solved complex structure of the protein with a 19 bp dsDNA had unveiled a left-handed helical superstructure just made up by DNA interactions. PMID- 11428898 TI - The crystal structure of Escherichia coli MoeA, a protein from the molybdopterin synthesis pathway. AB - MoeA is involved in synthesis of the molybdopterin cofactor, although its function is not yet clearly defined. The three-dimensional structure of the Escherichia coli protein was solved at 2.2 A resolution. The locations of highly conserved residues among the prokaryotic and eukaryotic MoeA homologs identifies a cleft in the dimer interface as the likely functional site. Of the four domains of MoeA, domain 2 displays a novel fold and domains 1 and 4 each have only one known structural homolog. Domain 3, in contrast, is structurally similar to many other proteins. The protein that resembles domain 3 most closely is MogA, another protein required for molybdopterin cofactor synthesis. The overall similarity between MoeA and MogA, and the similarities in a constellation of residues that are strongly conserved in MoeA, suggests that these proteins bind similar ligands or substrates and may have similar functions. PMID- 11428899 TI - Crystal structure of pentaerythritol tetranitrate reductase: "flipped" binding geometries for steroid substrates in different redox states of the enzyme. AB - Pentaerythritol tetranitrate reductase (PETN reductase) degrades high explosive molecules including nitrate esters, nitroaromatics and cyclic triazine compounds. The enzyme also binds a variety of cyclic enones, including steroids; some steroids act as substrates whilst others are inhibitors. Understanding the basis of reactivity with cyclic enones requires structural information for the enzyme and key complexes formed with steroid substrates and inhibitors. The crystal structure of oxidised and reduced PETN reductase at 1.5 A resolution establishes a close structural similarity to the beta/alpha-barrel flavoenzyme, old yellow enzyme. In complexes of oxidised PETN reductase with progesterone (an inhibitor), 1,4-androstadiene-3,17-dione and prednisone (both substrates) the steroids are stacked over the si-face of the flavin in an orientation different from that reported for old yellow enzyme. The specifically reducible 1,2 unsaturated bonds in 1,4-androstadiene-3,17-dione and prednisone are not optimally aligned with the flavin N5 in oxidised enzyme complexes. These structures suggest either relative "flipping" or shifting of the steroid with respect to the flavin when bound in different redox forms of the enzyme. Deuterium transfer from nicotinamide coenzyme to 1,4-androstadiene-3,17-dione via the enzyme bound FMN indicates 1alpha addition at the steroid C2 atom. These studies rule out lateral motion of the steroid and indicate that the steroid orientation is "flipped" in different redox states of the enzyme. PMID- 11428900 TI - Structure, dynamics and electrostatics of the active site of glutaredoxin 3 from Escherichia coli: comparison with functionally related proteins. AB - The chemistry of active-site cysteine residues is central to the activity of thiol-disulfide oxidoreductases of the thioredoxin superfamily. In these reactions, a nucleophilic thiolate is required, but the associated pK(a) values differ vastly in the superfamily, from less than 4 in DsbA to greater than 7 in Trx. The factors that stabilize this thiolate are, however, not clearly established. The glutaredoxins (Grxs), which are members of this superfamily, contain a Cys-Pro-Tyr-Cys motif in their active site. In reduced Grxs, the pK(a) of the N-terminal active-site nucleophilic cysteine residue is lowered significantly, and the stabilization of the corresponding thiolate is expected to influence the redox potential of these enzymes. Here, we use a combination of long molecular dynamics (MD) simulations, pK(a) calculations, and experimental investigations to derive the structure and dynamics of the reduced active site from Escherichia coli Grx3, and investigate the factors that stabilize the thiolate. Several different MD simulations converged toward a consensus conformation for the active-site cysteine residues (Cys11 and Cys14), after a number of local conformational changes. Key features of the model were tested experimentally by measurement of NMR scalar coupling constants, and determination of pK(a) values of selected residues. The pK(a) values of the Grx3 active-site residues were calculated during the MD simulations, and support the underlying structural model. The structure of Grx3, in combination with the pK(a) calculations, indicate that the pK(a) of the N-terminal active-site cysteine residue in Grx3 is intermediate between that of its counterpart in DsbA and Trx. The pK(a) values in best agreement with experiment are obtained with a low (<4) protein dielectric constant. The calculated pK(a) values fluctuate significantly in response to protein dynamics, which underscores the importance of the details of the underlying structures when calculating pK(a) values. The thiolate of Cys11 is stabilized primarily by direct hydrogen bonding with the amide protons of Tyr13 and Cys14 and the thiol proton of Cys14, rather than by long-range interactions from charged groups or from a helix macrodipole. From the comparison of reduced Grx3 with other members of the thioredoxin superfamily, a unifying theme for the structural basis of thiol pK(a) differences in this superfamily begins to emerge. PMID- 11428901 TI - Native-like beta-hairpin retained in the cold-denatured state of bovine beta lactoglobulin. AB - Bovine beta-lactoglobulin is denatured by increased temperature (heat denaturation) and by decreased temperature (cold-denaturation) in the presence of 4 M urea at pH 2.5. We characterized the structure of the cold-denatured state of beta-lactoglobulin using circular dichroism (CD), small-angle X-ray scattering (SAXS) and heteronuclear nuclear magnetic resonance (NMR). CD and SAXS indicated that the cold-denatured state, in comparison with the highly denatured state induced by urea, is rather compact, retaining some secondary structure, but no tertiary structure. The location of the residual structures in the cold-denatured state and their stability were characterized by 1H/2H exchange combined with heteronuclear NMR. The results indicated that the residues adjacent to the disulfide bond (C106-C119) connecting beta-strands G and H had markedly high protection factors, suggesting the presence of a native-like beta-hairpin stabilized by the disulfide bond. Since this beta-hairpin is conserved between different conformational states, including the kinetic refolding intermediate, it should be of paramount importance for the folding and stability of beta lactoglobulin. On the other hand, the non-native alpha-helix suggested for the folding intermediate was not detected in the cold-denatured state. The 1H/2H exchange experiments showed that the protection factors of a mixture of the native and cold-denatured states is strongly biased by that of the labile cold denatured state, consistent with a two-process model of the exchange. PMID- 11428902 TI - High enzymatic activity and chaperone function are mechanistically related features of the dimeric E. coli peptidyl-prolyl-isomerase FkpA. AB - We have recently described the existence of a chaperone activity for the dimeric peptidyl-prolyl cis/trans isomerase FkpA from the periplasm of Escherichia coli that is independent of its isomerase activity. We have now investigated the molecular mechanism of these two activities in vitro in greater detail. The isomerase activity with a protein substrate (RNaseT1) is characterized by a 100 fold higher k(cat)/K(M) value than with a short tetrapeptide substrate. This enhanced activity with a protein is due to an increased affinity towards the protein substrate mediated by a polypeptide-binding site that is distinct from the active site. The chaperone activity is also mediated by interaction of folding and unfolding intermediates with a binding site that is most likely identical to the polypeptide-binding site which enhances catalysis. Both activities are thus mechanistically related, being based on the transient interaction with this high-affinity polypeptide-binding site. Only the isomerase activity, but not the chaperone activity, with the substrate citrate synthase can be inhibited by FK520. Experiments with the isolated domains of FkpA imply that both the isomerase and the chaperone site are located on the highly conserved FKBP domain. The additional amino-terminal domain mediates the dimerization and thus places the two active sites of the FKBP domains in juxtaposition, such that they can simultaneously interact with a protein, and this is required for full catalytic activity. PMID- 11428903 TI - A study on the prevention and treatment of myopia with nacre on chicks. AB - The contents of mineral elements and amino acids in the hydrolysate of the traditional Chinese mineral medicine nacre have been determined. It has long since been proved by the practice of doctors of traditional Chinese medicine that pearl can be used to treat eye diseases. Based on such an understanding, a study is made of the influence of the said medicine on the incidence of myopia. First a form-sense-deprived model (FDM) for chicks is developed and the effect of the said medicine on the elongation of axis oculi is determined with an Abbe's comparator and an A-mode ultrasound instrument. The activity of superoxide dismutase (SOD), nitric oxide synthetase (NOS), and the content of nitric oxide (NO) in the retino-pigmental epithelium choroid homogenate are also analysed. The role of the said traditional Chinese mineral medicine in preventing and treating myopia is explained with respect to the above findings. The results obtained will provide a basis for using nacre, a traditional Chinese mineral medicine, to prevent and treat myopia. PMID- 11428904 TI - Effects of melatonin on enzyme activities of glucose-6-phosphate dehydrogenase from human erythrocytes in vitro and from rat erythrocytes in vivo. AB - The in vivo and in vitro effects of melatonin on glucose-6-phosphate dehydrogenase (G6PD) from human erythrocytes have been investigated. For this purpose, human erythrocyte glucose-6-phosphate dehydrogenase was purified, at the beginning, 13.654 times in a yield of 28% by using ammonium sulphate precipitation and 2',5'-ADP Sepharose 4B affinity gel. A temperature of +4 degrees C was maintained during the purification process. Enzyme activity was determined by the Beutler method using a spectrophotometer at 340 nm. This method was utilized for all kinetic studies. For in vitro experiments, the enzyme activity increased below 0.08 mM melatonin concentration and reached a plateau above 0.1 mM. Ten mg kg(-1)melatonin was administered intraperitonally and indicated the stimulatory effect on the enzyme. Time-dependent in vivo studies were executed for melatonin in Sprague-Dawley type rats. It was found that G6PD activity in the erythrocytes was increased by the melatonin in 1.5 and 3.5 h. These results show that both in vitro(below 0.08 mM) and in vivo pharmacological levels of melatonin increased enzyme activity in erythrocytes. The findings also indicate that melatonin may be pharmacologically useful in patients where a deficiency of the enzyme in red blood cells (RBC) causes haemolytic anaemia. PMID- 11428905 TI - d,l-sotalol enhances baroreflex sensitivity in conscious rats surviving acute myocardial infarction. AB - The aim of the present study was to characterize the influence of D,L-sotalol on arterial baroreflex sensitivity (BRS) which is generally considered as an estimate of vagal activity and has prognostic value in patients after myocardial infarction (MI). Conscious rats were studied 3 days after left anterior descending coronary artery ligation (n= 5) or sham-operation (SH, n= 6). BRS was determined by linear regression analysis of the RR-interval (interval between heart beats) and mean arterial pressure changes evoked by i.v. bolus injections of methoxamine (inducing reflex bradycardia, RB) and nitroprusside (inducing reflex tachycardia, RT). In MI-rats heart rate and mean arterial pressure were not significantly different from values in SH-rats, left ventricular end diastolic pressure was increased and contractility was depressed. The BRS (RB: MI: 0.48 +/- 0.04(*), SH: 0.79 +/- 0.08; RT: MI: 0.41 +/- 0.05(*), SH: 0.86 +/- 0.08 ms mmHg(-1)) ((*)P< 0.05 vs SH) was markedly reduced. d, l -Sotalol (1.5 mg kg(-1)i.v.) reduced heart rate (MI: -11 +/- 3 %(*), SH: -11 +/- 3 %(*)) and mean arterial pressure only moderately [MI: -6 +/- 4 %(n.s.), SH: -7 +/- 2 %(*)], while BRS depression in MI-rats was completely neutralized [RB: MI: 1.08 +/- 0.14(*), SH: 1.19 +/- 0.11(*); RT: MI: 0.84 +/- 0.08(*), SH: 0.88 +/- 0.12 (n.s.) ms mmHg(-1)] ((*)P< 0.05 vs pretreatment). The BRS is reduced in rats early after MI, indicating a depressed reflex vagal activity. Treatment with D,L-sotalol at a dose with little effect on heart rate and mean arterial pressure markedly enhances and, thus, restores BRS in MI-rats. These data suggest that D,L-sotalol has both peripheral and central effects leading to an increase of reflex vagal control of heart rate in rats. PMID- 11428906 TI - Endotoxin-induced vascular hyporesponsiveness in rat aorta: in vitro effect of aminoguanidine. AB - The current study was designed to evaluate the endotoxin-induced alterations of the mechanisms involved in Ca(2+)handling within the rat thoracic aorta and further to examine whether in vitro inhibition of inducible nitric oxide synthase (iNOS) by aminoguanidine would account for this effect or not. Endothelium denuded aortic rings from rats injected with lipopolysaccharide (LPS) (5 mg kg( 1), i.p. 18 h prior to functional studies) or saline were mounted in isolated organ baths. Various experimental conditions were studied on paired rings of the same animal which were incubated in the presence or absence of aminoguanidine (100 microM). Phenylephrine contractility in Ca(2+)-containing buffer or in Ca(2+)-free buffer, contractions induced by K(+)depolarization and CaCl(2)in depolarized muscle and by caffeine exposure were significantly decreased in LPS treated rings and were reversed by aminoguanidine exposure. Aminoguanidine also improved the contractions recorded while switching the Ca(2+)-free buffer to Ca(2+)-containing buffer. We conclude that endotoxin induces a generalized contractile defect in vascular smooth muscle including impairment in the influx of extracellular Ca(2+)and release of Ca(2+)from intracellular stores. An increase in iNOS activation leading to excessive nitric oxide synthesis, possibly non-endothelial in origin, may account for this defect. PMID- 11428907 TI - MDMA ("ecstasy") exhibits an anxiogenic-like activity in social encounters between male mice. AB - Recent studies have suggested that 3,4-methylenedioxymethamphetamine (MDMA; "ecstasy"), a synthetic amphetamine derivative, might exhibit an anxiogenic-like activity in rodents. In this work, we examined the effects of MDMA (1, 8 and 15 mg kg(-1), i.p.) on social encounters between male mice using an ethopharmacological approach. Likewise, we compared the behavioural profile of MDMA with that induced by FG 7142 (15 mg kg(-1), i.p.), an anxiogenic benzodiazepine receptor ligand. Individually housed mice were exposed to anosmic standard opponents 30 min after drug administration, and the encounters were videotaped and evaluated using an ethologically based analysis. MDMA (8 and 15 mg kg(-1)) produced a behavioural pattern characterized by a marked decrease of aggression (threat and attack) as well as social investigation, body care and digging behaviours, without affecting immobility. Moreover, avoidance/flee and defence/submission behaviours were significantly increased by the drug. A very similar behavioural profile was observed in FG 7142-treated animals. Overall, these findings might indicate that MDMA has anxiogenic-like properties in male mice. PMID- 11428908 TI - The role of enzyme induction and inhibition on cypermethrin hepatotoxicity. AB - Cypermethrin at different concentrations (100, 200, 400 and 800 ng ml(-1)) was incubated with a primary culture of rat hepatocytes. Cypermethrin was cytotoxic to rat hepatocytes at concentrations of 200 ng ml(-1)or greater. Toxicity was measured by a decrease in cell viability and leakage of ALT and AST enzymes into the culture medium. The role of cytochrome P450 in the hepatotoxicity of cypermethrin insecticide was investigated in fresh hepatocytes isolated either from phenobarbital pretreated rats or control rats and coincubated with SKF525A. Pretreatment with phenobarbital strongly protected the hepatocytes against the cypermethrin induced loss of cell viability percentage and increased enzyme leakage percentage. Coincubation of the hepatocytes with SKF525A, a well-known cytochrome P450 inhibitor, substantially potentiated the effect of cypermethrin on cell viability and enzyme leakage. These results suggest that the cytocidal hepatotoxicity of cypermethrin in primary hepatocyte culture depends on its parent compound and phenobarbital, as a cytochrome P450 inducer, could be of therapeutic value. PMID- 11428909 TI - Ethanol combined with cocaine inhibits amylase release in guinea pig pancreatic lobules. AB - Concurrent ingestion of alcohol and cocaine is a common occurrence in cocaine dependent individuals. Cocaethylene is a pharmacologically active metabolite of cocaine that is formed in the liver in the presence of ethanol. The effects of ethanol combined with cocaine on the exocrine pancreas are not known. We studied the effect of ethanol and cocaine, alone or in combination, and cocaethylene on amylase release from isolated lobules of the guinea pig pancreas. Incubation of lobules with ethanol plus cocaine produced a more evident reduction of amylase release than each drug alone. An even larger reduction was observed with cocaethylene. HPLC analysis of incubation medium showed that no cocaethylene was formed in vitro in the presence of ethanol and cocaine. It is concluded that cocaethylene could strongly contribute to inhibition of exocrine pancreatic secretion in individuals who coadminister alcohol with cocaine. PMID- 11428910 TI - Increased potency of some substituted short peptide analogues in comparison to galanin(1-15)-NH(2)in rat fundus strips. AB - The activity of short porcine galanin (Gal) analogues was tested in vitro using rat gastric fundus strips. The peptides contracted longitudinal smooth muscle in a concentration-dependent manner with the following order of potency: Gal(1-29) >[Cit(14)]Gal(1- 15) >[Asp(14)]Gal(1- 15) >[Dab(14)]Gal(1- 15) >[Nle(14)] Gal(1 15) >[Dpr(14)]Gal(1- 15) >[Arg(14)]Gal(1- 15) >[Orn(14)]Gal(1- 15) >Gal(1-15). Only in the case of two peptides, namely [Cit(14)]Gal(1-15) and [Dab(14)]Gal(1 15) did the values of Hill coefficients, estimated from the appropriate concentration-contraction curves, differ significantly from unity. Our results indicate that both N- and C-terminals of Gal molecule contribute towards the affinity and activity of Gal in rat gastric smooth muscle cell receptors, indicating that their integrity is essential for its full excitatory myogenic action. The substitution of histidine with citruline, aspartic acid, norleucine or diaminobutyric acid in position 14 of the amino acid chain led to a considerable increase in potency, suggesting that amino acids located at this position might play a crucial role where the strength of short analogues is concerned. PMID- 11428911 TI - The effects of ovariectomy and female sex hormones on hepatic metallothionein-I gene expression after injection of cadmium chloride in mice. AB - Metallothioneins (MTs) have a low molecular weight and have been considered to be important metal-binding proteins in defense from cadmium (Cd) toxicity in animals. These proteins are known to be induced by the injection of heavy metals such as Cd. Previously, we developed the measurement of the MT mRNA expression by the RT-PCR method. In this study, to clarify the relation between Cd-induced MT-I mRNA expression and female sex hormones in liver, we investigated the influences of the ovariectomy and female sex hormones on hepatic MT-I mRNA expression after Cd injection, and also investigated the effects of aging on hepatic MT-I mRNA expression in mice after Cd injection. We analysed the MT-I mRNA expression by the RT-PCR method. Cd-induced MT-I mRNA expression in ovariectomized mice was more than that in sham-operated mice (9 weeks old). Both 17 beta -estradiol and progesterone reduced the Cd-induced MT-I mRNA expression in ovariectomized mice (9 weeks old). Moreover, the MT-I mRNA expression in male mice was more than that of females (9 weeks old). However, the sex difference in the gene expression was not found in younger (4 weeks old) or older (46 weeks old) mice. These results suggest that the expression of hepatic MT-I mRNA after Cd injection is influenced by female sex hormones. PMID- 11428913 TI - Phylogeny of multidrug transporters. AB - We currently recognize five large ubiquitous superfamilies and one small eukaryotic-specific family in which cellular multidrug efflux pumps occur. One, the ABC superfamily, includes members that use ATP hydrolysis to drive drug efflux, but the MFS, RND, MATE and DMT superfamilies include members that are secondary carriers, functioning by drug:H(+)or drug:Na(+)antiport mechanisms. The small MET family seems to be restricted to endosomal membranes of eukaryotes, and only a single such system has been functionally characterized. In this review article, these families of drug transporters are discussed and evaluated from phylogenetic standpoints. PMID- 11428914 TI - Preventing drug access to targets: cell surface permeability barriers and active efflux in bacteria. AB - Bacteria, being unicellular, are constantly exposed to toxic compounds in their environment. Gram-negative bacteria and mycobacteria are unusually successful in surviving in the presence of toxic compounds because they combine two mechanisms of resistance. They produce effective permeability barriers, comprising the outer membrane and the mycolate-containing cell wall, on the cell surface. Further, they actively pump out drug molecules that trickle through the barrier, often utilizing multidrug efflux pumps. In Gram-negative bacteria, multidrug pumps of exceptionally wide specificity frequently interact with outer membrane channels and accessory proteins, forming multisubunit complexes that extrude drug molecules directly into the medium, bypassing the outer membrane barrier. PMID- 11428915 TI - Transcriptional regulation of multidrug efflux pumps in bacteria. AB - As integral membrane proteins demonstrating an extraordinarily wide substrate range, some degree of regulatory control over the expression of bacterial multidrug-resistance (MDR) transporters is to be expected. Excessive expression could be deleterious, due to direct, physical disruption of membrane integrity, or the unwanted export of essential metabolites, a potential side-effect of their broad substrate specificity. There are limited clues as to the physiological functions of most MDR transporters, but their expression is likely to be up regulated in response to the presence of natural substrates of these pumps. Thus, it is no surprise that MDR genes are subject to regulation at the local level, consisting of examples of both transcriptional repression and activation by proteins encoded adjacent to that for the transporter. Furthermore, an increasing number of MDR genes have also been found to be controlled by global transcriptional activator proteins. PMID- 11428916 TI - Towards the molecular mechanism of prokaryotic and eukaryotic multidrug transporters. AB - Due to their ability to extrude structurally dissimilar cytotoxic drugs out of the cell, multidrug transporters are able to reduce the cytoplasmic drug concentration, and, hence, are able to confer drug resistance on human cancer cells and pathogenic microorganisms. This review will focus on the molecular properties of two bacterial multidrug transporters, the ATP-binding cassette transporter LmrA and the proton motive force-dependent major facilitator superfamily transporter LmrP, which each represent a major class of multidrug transport proteins encountered in pro- and eukaryotic cells. In spite of the structural differences between LmrA and LmrP, the molecular bases of their drug transport activity may turn out to be more similar than might currently appear. PMID- 11428917 TI - Molecular genetic analysis and biochemical characterization of mammalian P glycoproteins involved in multidrug resistance. AB - A variety of human cancers become resistant or are intrinsically resistant to treatment with conventional drug therapies. This phenomenon is due in large part to the overexpression of a 170 kDa plasma membrane ATP-dependent pump known as the multidrug resistance transporter or P-glycoprotein. P-glycoprotein is a member of the large ATP binding cassette (ABC) superfamily of membrane transporters. This review focuses on the use of structure-function analyses to elucidate further the mechanism of action of mammalian P-glycoproteins. Ultimately, a complete understanding of the mechanism is important for the development of novel strategies for the treatment of many human cancers. PMID- 11428918 TI - Exploring the structure and function of the P-glycoprotein multidrug transporter using fluorescence spectroscopic tools. AB - P-glycoprotein is an ABC protein that functions as an efflux pump for multiple drugs, natural products and peptides. It is proposed to operate as a hydrophobic vacuum cleaner, expelling non-polar compounds from the membrane bilayer to the exterior, driven by the energy of ATP hydrolysis. The nucleotide-binding domains of P-glycoprotein appear to operate by an alternating sites mechanism to power drug transport. In recent years, purification and functional reconstitution of the protein has allowed the application of fluorescence spectroscopic techniques. This approach has led to insights into the structural architecture of the P glycoprotein molecule, and a more detailed understanding of the way in which it interacts with nucleotides and drugs. PMID- 11428919 TI - From models to molecules: opioid receptor dimers, bivalent ligands, and selective opioid receptor probes. PMID- 11428920 TI - Isomerization of (Z,Z) to (E,E)1-bromo-2,5-bis-(3-hydroxycarbonyl-4 hydroxy)styrylbenzene in strong base: probes for amyloid plaques in the brain. AB - In developing probes for detecting beta-amyloid (Abeta) plaques in the brain of Alzheimer's disease (AD), we have synthesized 1-bromo-2,5-bis-(3-hydroxycarbonyl 4-hydroxy)styrylbenzene (5, BSB). Due to the presence of two double bonds, formation of four different isomers is possible. Four isomers, E,E-5, E,Z-5, Z,E 5, and Z,Z-5, were prepared. Surprisingly, all showed strong fluorescent labeling of Abeta plaques in the brain of postmortem brain sections of patients with confirmed AD. In vitro binding assay also showed that all four isomers of BSB (E,E-5, E,Z-5, Z,E-5, and Z,Z-5) displayed a similar high binding affinity inhibiting the binding of [(125)I]E,E-6, 1-iodo-2,5-bis-(3-hydroxycarbonyl-4 methoxy)styrylbenzene (IMSB) to Abeta(1-40) aggregates. The inhibition constants (K(i)) of E,E-5, E,Z-5, Z,E-5, and Z,Z-5 were 0.11 +/- 0.01, 0.19 +/- 0.03, 0.27 +/- 0.06, and 0.13 +/- 0.02 nM, respectively. Due to the fact that geometric stability of these styrylbenzenes is unknown, and the conversion of Z,Z-5 to E,E 5 may occur automatically in the binding or labeling assaying conditions, we have investigated the kinetics of conversion of Z,Z-5 to E,E-5 by NMR in D(2)O/NaOD at elevated temperatures (70, 95, and 115 degrees C). The activation energy was determined to be 14.15 kcal/mol. The results strongly suggest that the isomeric conversion at room temperature in aqueous buffer solution is unlikely. All of the styrylbenzene isomers clearly showed potential as useful tools for studying Abeta aggregates in the brain. The data suggest that, despite the rigidity of this series of styrylbenzenes, the binding sites on Abeta aggregates may have certain flexibility and the binding pockets could be adaptable for binding to other smaller ligands. Such information could be exploited to develop new ligands for detecting amyloid plaques in AD. PMID- 11428921 TI - Utilization of an intramolecular hydrogen bond to increase the CNS penetration of an NK(1) receptor antagonist. AB - This paper describes the synthesis and physical and biological effects of introducing different substituents at the alpha-position of the tryptophan containing neurokinin-1 receptor antagonist [(R)-2-(1H-indol-3-yl)-1-methyl-1 ((S)-1-phenyl-ethylcarbamoyl)-ethyl]-carbamic acid benzofuran-2-ylmethyl ester (CI 1021). The described compounds all exhibit less than 5 nM binding affinities for the human neurokinin-1 receptor and selectivity over the tachykinin NK(2) and NK(3) receptor subtypes. Application of variable temperature nuclear magnetic resonance spectroscopy studies of the amide and urethane protons was utilized to determine the existence of an intramolecular hydrogen bond. This intramolecular hydrogen bond increases the apparent lipophilicity to allow increased central nervous system penetration and pharmacological activity (gerbil foot tap test) in the case of the highest affinity compound [(S)-1-dimethylaminomethyl-2-(1H-indol 3-yl)-1-((S)-1-phenyl-ethylcarbamoyl)-ethyl]-carbamic acid benzofuran-2-ylmethyl ester (PD 174424) over those analogues that could not form an intramolecular hydrogen bond. PMID- 11428922 TI - Novel N-(arylalkyl)indol-3-ylglyoxylylamides targeted as ligands of the benzodiazepine receptor: synthesis, biological evaluation, and molecular modeling analysis of the structure-activity relationships. AB - A series of N-(arylalkyl)indol-3-ylglyoxylylamides (4-8) was synthesized as ligands of the benzodiazepine receptor (BzR) and tested for their ability to displace [(3)H]flumazenil from bovine brain membranes. The new compounds, bearing a branched (4) or a geometrically constrained benzyl/phenylethyl amide side chain (5-8), represent the continuation of our research on N-benzylindol-3 ylglyoxylylamides 1 (Da Settimo et al., 1996), N'-phenylindol-3 ylglyoxylohydrazides 2 (Da Settimo et al., 1998), and N-(indol-3 ylglyoxylyl)alanine derivatives 3 (Primofiore et al., 1989). A few indoles belonging to the previously investigated benzylamides 1 and phenylhydrazides 2 were synthesized and tested to enrich the SARs in these two series. The affinities and the GABA ratios of selected compounds for clonal mammalian alpha(1)beta(2)gamma(2), alpha(3)beta(2)gamma(2), and alpha(5)beta(3)gamma(2) BzR subtypes were also determined. It was hypothesized that the reduced flexibility of indoles 4-8 would both facilitate the mapping of the BzR binding cleft and increase the chances of conferring selectivity for the considered receptor subtypes. In the series of indoles 4, the introduction of a methyl group on the benzylic carbon with the R configuration improved affinity of the 5-substituted (5-Cl and 5-NO(2)) derivatives, whereas it was detrimental for their 5 unsubtituted (5-H) counterparts. All S enantiomers were less potent than the R ones. Replacement of the methyl with hydrophilic substituents on the benzylic carbon lowered affinity. The isoindolinylamide side chain was tolerated if the 5 position was unsubstituted (K(i) of 5a = 123 nM), otherwise affinity was abolished (5b, c). All the 2-indanylamides 6 and (S)-1-indanylamides 8 were devoid of any appreciable affinity. The 5-Cl and 5-NO(2) (R)-1-indanylamides 7b (K(i) 80 nM) and 7c (K(i) 28 nM) were the most potent among the indoles 5-8 geometrically constrained about the side chain. The 5-H (R)-1-indanylamide 7a displayed a lower affinity (K(i) 675 nM). The SARs developed from the new compounds, together with those collected from our previous studies, confirmed the hypothesis of different binding modes for 5-substituted and 5-unsubstituted indoles, suggesting that the shape of the lipophilic pocket L(1) (notation in accordance with Cook's BzR topological model) is asymmetric and highlighted the stereoelectronic and conformational properties of the amide side chain required for high potency. Several of the new indoles showed selectivity for the alpha(1)beta(2)gamma(2) subtype compared with the alpha(3)beta(2)gamma(2) and alpha(5)beta(3)gamma(2) subtypes (e.g.: 4t and 7c bind to these three BzR isoforms with K(i) values of 14 nM, 283 nM, 239 nM, and 9 nM, 1960 nM, 95 nM, respectively). The GABA ratios close to unity exhibited by all the tested compounds on each BzR subtype were predictive of an efficacy profile typical of antagonists. PMID- 11428923 TI - Enantioselective syntheses of potent retinoid X receptor ligands: differential biological activities of individual antipodes. AB - The synthesis and characterization of chiral RXR selective ligands are described. The enantiomeric acids 2 and 3 were synthesized employing an enantioselective cylopropanation procedure as the key step. Compound 2, with an S,S configuration at C-9 and C-10, is a potent RXR agonist devoid of any RAR activity. The R,R enantiomer 3 is a weak RXR agonist and has demonstrable RAR activity in the receptor transactivation assays. The potent RXR activity of 2 was further confirmed in a hyperglycemic animal model (db/db mice). Compound 2 lowered glucose by 50% by day 7 at 2 mg/kg, whereas 3 had no effect at the same dosage. This further supports the contention that RXR mediated gene transcription is involved in the antidiabetic effects of RXR ligands. PMID- 11428924 TI - Discovery of novel mesangial cell proliferation inhibitors using a three dimensional database searching method. AB - A three-dimensional pharmacophore model of mesangial cell (MC) proliferation inhibitors was generated from a training set of 4-(diethoxyphosphoryl)methyl-N-(3 phenyl-[1,2,4]thiadiazol-5-yl)benzamide, 2, and its derivatives using the Catalyst/HIPHOP software program. On the basis of the in vitro MC proliferation inhibitory activity, a pharmacophore model was generated as seven features consisting of two hydrophobic regions, two hydrophobic aromatic regions, and three hydrogen bond acceptors. Using this model as a three-dimensional query to search the Maybridge database, structurally novel 41 compounds were identified. The evaluation of MC proliferation inhibitory activity using available samples from the 41 identified compounds exhibited over 50% inhibitory activity at the 100 nM range. Interestingly, the newly identified compounds by the 3D database searching method exhibited the reduced inhibition of normal proximal tubular epithelial cell proliferation compared to a training set of compounds. PMID- 11428925 TI - Heterocycle-containing retinoids. Discovery of a novel isoxazole arotinoid possessing potent apoptotic activity in multidrug and drug-induced apoptosis resistant cells. AB - In a search for retinoic acid (RA) receptor ligands endowed with potent apoptotic activity, a series of novel arotinoids were prepared. Because the stereochemistry of the C9-alkenyl portion of natural 9-cis-RA and the olefinic moiety of the previously synthesized isoxazole retinoid 4 seems to have particular importance for their apoptotic activity, novel retinoid analogues with a restricted or, vice versa, a larger flexibility in this region were designed and prepared. The new compounds were evaluated in vitro for their ability to activate natural retinoid receptors and for their differentiation-inducing activity. Cytotoxic and apoptotic activities were, in addition, evaluated. In general, these analogues showed low cytotoxicity, with the restricted structures being slightly more active than the more flexible ones. As an exception, however, the isoxazole retinoid 15b proved to be particularly able to induce apoptosis at concentrations <5 microM, showing a higher activity than the classical retinoids such as all trans-RA, 13-cis-RA, and 9-cis-RA and the previously described synthetic retinoid 4. 15b also exhibited a good affinity for the retinoid receptors. Interestingly, another important property of 15b was its ability to induce apoptosis in the HL60R multidrug-resistant (MDR) cell line, at the same concentration as is effective in HL60. Therefore, 15b represents a new retinoid possessing high apoptotic activity in an MDR cell line. The ability of 15b to act on K562 and HL60R cells suggests that this compound may have important implications in the treatment of different leukemias, and its structure could offer an interesting model for the design of new compounds endowed with apoptotic activity on MDR- and retinoid-resistant malignancies. PMID- 11428926 TI - 4-Hydroxy-5,6-dihydropyrones as inhibitors of HIV protease: the effect of heterocyclic substituents at C-6 on antiviral potency and pharmacokinetic parameters. AB - Due largely to the emergence of multi-drug-resistant HIV strains, the development of new HIV protease inhibitors remains a high priority for the pharmaceutical industry. Toward this end, we previously identified a 4-hydroxy-5,6-dihydropyrone lead compound (CI-1029, 1) which possesses excellent activity against the protease enzyme, good antiviral efficacy in cellular assays, and promising bioavailability in several animal species. The search for a suitable back-up candidate centered on the replacement of the aniline moiety at C-6 with an appropriately substituted heterocyle. In general, this series of heterocyclic inhibitors displayed good activity (in both enzymatic and cellular tests) and low cellular toxicity; furthermore, several analogues exhibited improved pharmacokinetic parameters in animal models. The compound with the best combination of high potency, low toxicity, and favorable bioavailabilty was (S)-3 (2-tert-butyl-4-hydroxymethyl-5-methyl-phenylsulfanyl)-4-hydroxy-6-isopropyl-6-(2 thiophen-3-yl-ethyl)-5,6-dihydro-pyran-2-one (13-(S)). This thiophene derivative also exhibited excellent antiviral efficacy against mutant HIV protease and resistant HIV strains. For these reasons, compound 13-(S) was chosen for further preclinical evaluation. PMID- 11428927 TI - A new concept for multidimensional selection of ligand conformations (MultiSelect) and multidimensional scoring (MultiScore) of protein-ligand binding affinities. AB - In this work, eight different scoring functions have been combined with the aim of improving the prediction of protein-ligand binding conformations and affinities. The obtained scores were analyzed using multivariate statistical methods to generate expressions, with the ability (1) to select the best candidate between different docked conformations of an inhibitor (MultiSelect) and (2) to quantify the protein-ligand binding affinity (MultiScore). By use of the docking program GOLD, 40 different inhibitors were docked into the active site of three matrix metalloproteinases (MMP's), yielding a total of 120 enzyme inhibitor complexes. For each complex, a single conformation of the inhibitor was selected using principal component analysis (PCA) for the scores obtained by the eight functions SCORE, LUDI, GRID, PMF_Score, D_Score, G_Score, ChemScore, and F_Score. Binding affinities were estimated based on partial least-squares projections onto latent structures (PLS) on the eight scores of each selected inhibitor conformation. By use of this procedure, R(2) = 0.78 and Q(2) = 0.78 were obtained when comparing experimental and calculated binding affinities. MultiSelect was evaluated by applying the same method for selecting docked conformations for 18 different protein-ligand complexes of known three dimensional structure. In all cases, the selected ligand conformations were found to be very similar to the experimentally determined ligand conformations. A more general evaluation of MultiScore was performed using a set of 120 different protein-ligand complexes for which both the three-dimensional structures and the binding affinities were known. This approach allowed an evaluation of MultiScore independently of MultiSelect. The generality of the method was verified by obtaining R(2) = 0.68 and Q(2) = 0.67, when comparing calculated and experimental binding affinities for the 120 X-ray structures. In all cases, LUDI, SCORE, GRID, and F_Score were included as important functions, whereas the fifth function was PMF_Score and ChemScore for the MMP and X-ray models, respectively. PMID- 11428928 TI - Synthesis and structure-activity relationships of trisubstituted phenyl urea derivatives as neuropeptide Y5 receptor antagonists. AB - 1-((1R,2R)-2-Hydroxy-1-methyl-2-phenylethyl)-1-methyl-3-(4-phenoxyphenyl)urea (1) was identified as a hit from the screening of the neuropeptide Y5 (NPY5) receptor. This lead was optimized for in vitro potency by changing the stereochemistry, the phenylethyl segment, the urea portion, and the 4 phenoxyphenyl group on the molecule. Over 40 analogues of 1 were prepared to study the structure-activity relationship for this series. The most potent compounds in this class have IC(50)s less than 0.1 nM at the NPY5 receptor (e.g., 40f, 44a, and 47). To determine the functional activity for this series of compounds, selected analogues were tested in a cellular assay measuring forskolin induced cyclic AMP accumulation in 293 cells transfected with the human NPY5 receptor. All urea analogues tested in the functional assay acted as antagonists (e.g., 1, 32, 40a, and 44e). PMID- 11428929 TI - Synthesis and antimalarial activity of novel medium-sized 1,2,4,5 tetraoxacycloalkanes. AB - CsOH- or Ag(2)O-mediated cycloalkylation of (alkylidene)bisperoxides 3 and 1,n dihaloalkanes (n = 3-8) provided the corresponding medium-sized 1,2,4,5 tetraoxacycloalkanes 4-8 in moderate yields. Subsequent evaluation of the antimalarial activity of the cyclic peroxides 4-8 in vitro and in vivo revealed that 1,2,6,7-tetraoxaspiro[7.11]nonadecane 4a has considerable potential as a new, inexpensive, and potent antimalarial drug. PMID- 11428930 TI - Novel inhibitors of mitochondrial respiratory chain: endoperoxides from the marine tunicate Stolonica socialis. AB - The Mediterranean tunicate Stolonica socialis contains a new class of powerful cytotoxic acetogenins, generically named stolonoxides. In this paper, which also details the isolation and chemical characterization of a minor component (3a) of the tunicate extract, we report the potent inhibitory activity (IC(50) < 1 microM) of stolonoxides (1a and 3a) on mitochondrial electron transfer. The compounds affect specifically the functionality of complex II (succinate:ubiquinone oxidoreductase) and complex III (ubiquinol:cytochrome C oxidoreductase) in mammalian cells, thereby causing a rapid collapse of the whole energetic metabolism. This result, which differs from the properties of similar known products (e.g., 6), reflects the molecular features of stolonoxides. PMID- 11428931 TI - Synthesis and biological activity of 7-oxo substituted analogues of 5-deaza 5,6,7,8-tetrahydrofolic acid (5-DATHF) and 5,10-dideaza-5,6,7,8-tetrahydrofolic acid (DDATHF). AB - We recently described the syntheses of 12a-c, 4-amino-7-oxo substituted analogues of 5-deaza-5,6,7,8-tetrahydrofolic acid (5-DATHF), and 5,10-dideaza-5,6,7,8 tetrahydrofolic acid (DDATHF), in six steps from commercially available p substituted methyl benzoates in 20-27% overall yields. Such analogues were tested in vitro against CCRF-CEM leukemia cells and showed that they are completely devoid of any activity, the IC(50) being higher than 20 microg/mL for all cases. To clarify if the presence of the carbonyl group in position C7, the distinctive feature of our synthetic methodology, is the reason for this lack of activity, we have now obtained the 7-oxo substituted analogues of 5-DATHF and DDATHF, 18a-c, in 10-30% overall yield. Testing of 18a-c in vitro against CCRF-CEM leukemia cells revealed that these compounds are totally inactive. A molecular modeling study of 18b inside the active site of the complex E. coliGARTFase-5-DATHF-GAR pointed to an electronic repulsion between the atoms of the 7-oxo group and the carbonyl group of Arg90 as a possible explanation for the inactivity of 18a-c. PMID- 11428932 TI - Peptide analogues of a subdominant epitope expressed in ebv-associated tumors: synthesis and immunological activity. AB - H-Cys-Leu-Gly-Gly-Leu-Leu-Thr-Met-Val-OH (CLG) peptide is an EBV subdominant epitope that represents the target of HLA-A2 restricted CTL responses. The CLG peptide has low affinity for HLA-A2 and does not produce stable complexes, both factors that determine weak CTL responses. In contrast, the [Tyr(1), Ala(3)]CLG (YLA) analogue showed high affinity for HLA-A2 molecules and efficiently stimulated CLG-specific CTL precursors. Nevertheless, this modified epitope showed low enzymatic stability. To further improve the immunotherapeutical potential of this "improved epitope", we have synthesized and tested YLA analogues containing different modifications next to the scissile peptide bond. Among the analogues we found three peptides, with higher enzymatic resistance, that efficiently stimulate CTL responses. These peptides may be used for EBV specific immunotherapies. PMID- 11428933 TI - Synthesis and antibacterial evaluation of certain quinolone derivatives. AB - A number of 7-substituted quinolone derivatives were synthesized and evaluated for antibacterial and cytotoxic activities. Preliminary results indicated that most compounds tested in this study demonstrated better activity against methicillin-resistant Staphylococcus aureus than norfloxacin. Among them, 1-(4 amino-2-fluorophenyl)-6-fluoro-1,4-dihydro-7-[4-[2-(4-methoxyphenyl)-2 hydroxyiminoethyl]-1-piperazinyl]-4-oxo-3-quinolinecarboxylic acid (11d) and its ketone precursor 10d exhibited significant activities against Klebsiella pneumoniae, methicillin-resistant S. aureus, erythromycin- and ampicillin resistant Streptococcus pneumoniae, and vancomycin-resistant Enterococcus faecalis. Due to strong cytotoxicities of 11d (a mean log GI(50) of -5.40), compound 10d, with good antibacterial activities and low cytotoxicities (a mean log GI(50) of -4.67), is a more potential drug candidate. PMID- 11428934 TI - Conformational analysis of a glycosylated human myelin oligodendrocyte glycoprotein peptide epitope able to detect antibody response in multiple sclerosis. AB - Myelin oligodendrocyte glycoprotein (MOG), a minor myelin component, is an important central nervous system specific target autoantigen for primary demyelination in autoimmune diseases such as multiple sclerosis (MS). The native structure of MOG presents a glycosylation site at position 31 (Asn(31)). It has been recently described that glycosylation of a MOG peptide epitope improved the detection of specific autoantibodies in sera of MS patients. The solution conformational behavior of two MOG derived peptides-hMOG(30-50) (1) and the glycosylated analogue [Asn(31)(N-beta-Glc)]hMOG(30-50) (2)-were investigated through NMR analysis in a water/HFA solution. Conformational studies revealed that peptides 1 and 2 adopted similar conformations in this environment. In particular, they showed strong propensity to assume a well-defined amphipatic structure encompassing residues 41-48. The N-terminal region resulted to be almost completely unstructured for both peptides. The presence in 1 of a low populated Asx-turn conformation characteristic of the Asn-Xaa-Thr glycosylation sites was the only conformational difference between peptides 1 and 2. Thus, the specific antibody recognition of peptide 2 is most likely driven by direct interactions of the antibody binding site with the Asn-linked sugar moiety. PMID- 11428935 TI - Automated melanoma diagnosis: where are we at? AB - BACKGROUND: It has been over a decade since the first mention was made of the computer as a tool for assisting clinicians in diagnosing skin lesions. This review tabulates and summarises the major research papers, and comments on the state of the field after a decade of research. CONCLUSIONS: We conclude that epiluminescent microscopy has become the image-capture technique of choice in this field. However, the reporting of research to date has been less than exemplary, making "reinvention of the wheel" likely. It also appears that although the goal of a clinically useful diagnostic system is closer, the complexity and variation displayed by skin lesions, coupled with the ad hoc direction and reporting of research, may hinder the achievement of this goal for some time to come. PMID- 11428936 TI - In vivo optical coherence tomography imaging of human skin: norm and pathology. AB - BACKGROUND/AIMS: Since the majority of skin diseases are known to be accompanied by structural alterations, research efforts are focused on the development of various novel diagnostic techniques capable of providing in vivo information on the skin structure. An essential parameter here is spatial resolution. In this paper we demonstrate the capabilities of optical coherence tomography (OCT) in detecting in vivo specific features of thin and thick skin. A particular focus is made on the identification of OCT patterns typical of certain pathological processes in skin, by performing parallel histological and tomographical studies. METHODS: To obtain images of the skin, we used a compact fiber OCT system developed at the Institute of Applied Physics of the Russian Academy of Sciences. A low coherence source (superluminescent diode) operated at a wavelength of 1280 nm; the output power was 0.5-2 mW. This power is low enough to conform to the ANSI safety standards for light exposure. The in-depth resolution limited by the spectral bandwidth (40-50 nm) of the probing light was approximately 20 &mgr;m. The lateral resolution determined by the probe light focusing ranged from 15 to 30 &mgr;m. In this series of experiments the maximum depth of imaging did not extend beyond 1.5 mm. Obtaining images of skin regions 2-6 mm long took 2-4 s. OCT capabilities for imaging normal skin of different localization and some skin diseases were studied in 12 healthy volunteers and 24 patients. RESULTS: OCT imaging of the skin can detect in vivo such general pathological reactions of the human body as active inflammation and necrosis. OCT is useful for in vivo diagnosis of some specific processes in the skin, including hyperkeratosis, parakeratosis and formation of intradermal cavities. OCT imaging is noninvasive and therefore allows frequent multifocal examination of skin without any adverse effects. OCT can perform monitoring of disease progress and recovery in the course of therapy. Morphometric studies, measurements of the depth and extension of skin pathology within the human body can be easily performed by OCT. CONCLUSIONS: OCT allows imaging of subsurface soft tissues with the spatial resolution of 15-20 &mgr;m, a resolution one order of magnitude higher than that provided by other clinically available noninvasive diagnostic techniques. An imaging depth of up to 1.5-2 mm, given by current OCT technology, is sufficient to examine the skin. Real time OCT imaging can provide information not only on the structure, but also on some specific features in the functional state, of tissues. OCT imaging is a noninvasive technique, i.e., OCT does not cause trauma and has no side effects since it utilizes radiation in the near infrared wavelength range at a power as low as 1 mW. PMID- 11428937 TI - Interdigitation index - a parameter for differentiating between young and older skin specimens. AB - BACKGROUND/AIMS: Quantitative morphometry developed rapidly during the last decade due to advances is computers and software. We wish to establish a simple baseline for the morphometric differences due to intrinsic ageing between young and old cohorts: the interdigitation index. It is an expression of the shape of the border between the epidermis and dermis. METHODS: We used volar forearm biopsies of women, since the volar forearm is usually not photodamaged. The biopsies were fixed in buffered formalin, embedded in paraffin and sectioned. Separate sections were stained by hematoxylin-eosin, by orcein and by dimethyl methylene blue. We had seven female volunteers in each group; the young cohort had a mean age of 26.6 years, the older cohort 50.9 years. We chose a cohort that was just about postmenopausal, since in the future we wish to evaluate the effect of externally-applied agents on postmenopausal female skin and the earlier it is applied the better its chance of being effective. RESULTS: We found no difference between the young and older cohort with regard to epidermal thickness. We found a decrease of glycosaminoglycen (GAG) as measured by dimethyl-methylene blue staining. The results of the elastic staining by orcein, although in line with the reports in the literature, are not useful for evaluating the intrinsic ageing process, at least not by the simple percentage of area stained procedure. We introduced a new parameter: the interdigitation index. It is a simple measurement of the interdigitation in the epidermal-dermal junction, known to be diminished by age. This index was diminished by about 20% between the young and older cohort. CONCLUSIONS: Quantitative morphometry using simple epidermal and dermal measurements on biopsies of the volar forearm of women is suitable for following intrinsic ageing of the skin and offers a simple objective method for following the ageing process of the skin. PMID- 11428938 TI - Potential of a high-frequency correlation method to study skin blood flow. AB - BACKGROUND/AIMS: The vast number of existing dedicated techniques proves that skin blood flow estimation is an unsolved problem. Specificities of cutaneous vascularization (very low blood velocity, noisy environment, complexity of the vascularization architecture) result in the unsuitability of conventional ultrasonic Doppler techniques (long acquisition time, low spatial resolution). The object here was to present a high-frequency time-domain correlation METHOD: In particular, the difficulties of adaptating this type of measurement (data processing, hardware problem) are pointed out. METHODS: Radio-frequency (RF) backscattered signals, obtained with a modified version of a home-made 20 MHz skin imaging system, are studied. Time shifts between successive windowed sections of the RF signals are determined by the mean of the cross-correlation algorithm. A realignment procedure (to remove the artefacts caused by the movements of the patient and the manipulator) and a stationary echo cancelling procedure (to remove the signals coming from the cutaneous tissues and to permit the detection of very small vessels) are used. RESULTS: In vitro results show that velocity measurements as low as 0.1 mm/s are attainable with a 80 &mgr;m axial resolution, and blood vessels of 100 &mgr;m are detectable. Our technique has also been validated by means of in vivo experiment on an erysipelas located on a human leg. In this way, a 180-&mgr;m-diameter blood vessel has been detected on a M-mode RF image and the corresponding velocity profile has been obtained. CONCLUSION: Further improvements can be expected, and the level of performance obtained in vitro in this work should be also attainable in vivo and should then provide an effective tool for skin physiology and pathology. PMID- 11428939 TI - Is vascular reactivity in skin predictable? AB - BACKGROUND/AIMS: Cutaneous microdialysis can be used to follow the pharmacology and metabolism of an inflammatory reaction. It is assumed that an equilibration period compensates for the trauma of catheter insertion. METHODS: In the present paper, the vascular reactivity of the skin to histamine was tested using assessment by laser Doppler perfusion imaging. In a group of six subjects, the provocation was preceded by insertion of a microdialysis catheter. In a second control group, histamine provocation alone was performed (six subjects). RESULTS: The vascular response to histamine was greater in the microdialysis catheter group than the control group. The histamine provocation caused a greater response than catheter insertion. Further, there was a correlation between the response to catheter insertion and the histamine provocation. CONCLUSION: It appears that the insertion and/or presence of a microdialysis catheter increases vascular reactivity to subsequent provocation. The response to catheter insertion may predict skin reactivity in general in the individual subject. PMID- 11428940 TI - Transepidermal water loss, temperature and sebum levels on women's facial skin follow characteristic patterns. AB - BACKGROUND/AIMS: The aim of this study was to compare the biophysical properties of different facial zones. METHODS: We investigated transepidermal water loss (TEWL), skin temperature and sebum casual level (CL) on 90 adjacent test sites distributed on the forehead, cheeks and chin of five women. RESULTS: All three parameters showed a symmetrical distribution around the facial median line. Only minor variations of individual values were found within the forehead and the chin areas. In contrast, the cheeks exhibited a distinct gradient with highest values in the paranasal zones and lowest on the cheek bones for all of the three parameters. The mean values on both cheeks of a given individual were nearly identical, and the patterns within the two cheeks were superimposable. Both CL and skin temperature distributions pointed out a "T-zone" with highest values on the forehead, on the chin and on the median part of the cheek. CONCLUSIONS: Our data demonstrate that biophysical skin properties differ considerably between different facial areas but that they follow a characteristic distribution. PMID- 11428941 TI - A novel in vivo model in guinea pigs for dry skin syndrome. AB - BACKGROUND/AIMS: The lack of a suitable, validated animal model for the comparison of the pharmacological effectiveness of known and potential moisturizers in the treatment of "dry skin syndrome" led us to develop such an in vivo model. METHODS: "Dry skin syndrome" was induced in guinea pigs by daily application of 2% sodium lauryl sulphate (SLS) in deionized water on one of the two shaved flanks for three consecutive days. After ascertaining skin dryness, that side was treated with an agent for 6 days. The in vivo humectant effect was measured by a Corneometer CM 825(R), erythema was measured by a Mexameter MX 16(R). In some cases histological studies were carried out. RESULTS: The treatment with the 2% SLS led to a consistent "dry skin syndrome" for 2 weeks. Glycerol, Vaseline, urea and ammonium lactate treatments validated the model, since the Corneometer CM 825(R) readings of the treated dry side was equal to that of the control untreated side after 1 week of treatment. Mexameter MX 16(R) measurements showed abolishment of the erythema by glycerol only. Histological study showed that SLS treatment creates acanthosis that is partially reversed by Vaseline and fully reversed by glycerol treatment. CONCLUSION: The guinea pig dry skin model is a relevant model of the human "dry skin syndrome". The instrumental results combined with the histological findings indicate that erythema measurements are relevant for the determination of curative effect. PMID- 11428942 TI - Imaging, image processing and pattern analysis of skin capillary ensembles. AB - BACKGROUND/AIMS: The capillary bed is recognized as the site where metabolic and nutrient processes occur for living tissues at all levels. The evaluation of this vital process is a major concern in microcirculation. Unlike traditional approaches that concentrated on the extreme local properties of this process, a more global analysis toward capillary ensembles is employed here, since capillaries work as a cooperative entirety. As a first step toward ensemble analysis, the static and planar geometric parameters are investigated. Parameters such as the capillary adjacency and size information are very important in predicting and analysing certain malfunctions in the microvascular bed. METHODS/RESULTS: In order to achieve an objective and accurate analysis of these vital parameters, a computerized imaging system is proposed. Not only the number of capillaries and the capillary cross-sectional areas are important in describing the microvascular bed but the planar distribution pattern of the capillaries also carries valid information. This information, unique to the ensemble analysis, can be used to reveal, visualise and quantify the clustering of capillaries; and this information, according to the Krogh model, is fundamental in estimating the tissue oxygen supply. Two spatial models, the closest neighbor and triangulation methods, have been applied to the captured images of capillary ensembles. The closest neighbor technique generates a minimal distance map or displays a distribution, which depicts the local clustering of capillaries. The triangulation technique, on the other hand, generates a mutual distance map, which is a global description of the capillary positions. Triangulation methods have been evaluated but all except the Greedy triangulation method have been rejected due to lack of robustness and model weakness. Therefore, the capillaries are triangulated by the Greedy triangulation method, and the capillary distribution uniformity is defined as one minus the coefficient of variance of the edge lengths of the mutual distance map. CONCLUSIONS: A series of advanced image processing methods have been developed that efficiently extract the capillary position, size and distribution information from the images. These results facilitate the automatic counting of capillaries and the capillary size related pathological analysis. PMID- 11428943 TI - Computer recognition of skin structures using discriminant and cluster analysis. AB - BACKGROUND/AIMS: Automated image analysis of complex tissues is usually limited by the difficulty of recognizing special structures by computer. The aim of this study was to test the applicability of discriminant and cluster analysis to the interpretation of skin images. METHODS: Digital images from microscopic, dermatoscopic and clinical views of skin specimens were electronically dissected into elements of equal size and shape, and a set of grey level, colour and texture features was assessed for each element. Elements were classified interactively and submitted to discriminant analysis. Furthermore, hierarchical cluster analysis was used to enable the system to classify the tissue elements automatically, based on the available digital information. The classification results were relocated to the original image in order to evaluate the performance of the procedure. RESULTS: The system performs well in reproducibly detecting different skin structures in digital images. Discriminant analysis of interactively classified elements yielded a correct reclassification in 98 to 100% of tissue elements. Among the cluster analysis procedures, the conservative Ward method after removal of all highly correlated features produced the best results. The method turned out to be applicable irrespective of the image source used. CONCLUSIONS: Discriminant and cluster analysis may be helpful techniques for a user-independent, subjectively unbiased measurement system of skin structures. PMID- 11428944 TI - Biophysical measurements of human forearm skin in vivo: effects of site, gender, chirality and time. AB - BACKGROUND/AIMS: Measurements of transepidermal water loss (TEWL) and skin colour are biophysical techniques commonly used to measure the in vivo skin effects of cosmetics, topical medicaments and chemical irritants. The purpose of this study was to investigate the variability of TEWL and skin colour on human forearm skin as a function of regional variation, gender and preferred chirality over an 8 h period. METHODS: Biophysical measurements of TEWL and skin colour were made at five sites on both forearms of male (n=8) and female (n=9) human volunteers in vivo (38% relative humidity, 21 degrees C). RESULTS: Rates of TEWL at the forearm midpoint were 10% lower than at the forearm extremities (P<0.01). Skin redness (a*) near the wrist was 5-10% higher than at other sites (P<0.05). Rates of TEWL were 5% higher in male volunteers (P<0.05). Red and blue (b*) colour measurements of male forearm skin differed by 18% and 20% in comparison with female, respectively. Rates of TEWL, skin brightness (L*) and b* decreased by 9% (P<0.05), 1.8% (P<0.05) and 4% (P<0.05), respectively, with time whereas a* and skin temperature increased by 4.5% (P<0.01) and 7.2% (P<0.01), respectively. There was a significant correlation between the change in all measured parameters with time. CONCLUSIONS: Significant differences in TEWL and skin colour were identified that may have relevance in the design and interpretation of multivariate analyses of human forearm skin. Diurnal variation of TEWL, skin colour and temperature may have a single underlying mechanism. PMID- 11428945 TI - Seasonal variability in the biophysical properties of stratum corneum from different anatomical sites. AB - BACKGROUND/PURPOSE: A 10-month-long study on a panel of 24 young female subjects was carried out to determine whether various biophysical aspects of the stratum corneum (SC) varied with season. METHODS: Three different anatomical sites (calf, inner forearm and crow's foot wrinkle area of the face) were assessed in February, April, July and December of the same year. The assessments made were skin surface hydration using an electrical capacitance technique, transepidermal water loss by evaporimetry, number of corneocytes released using a turbine stimulation method, and skin surface topography using optical profilometry. RESULTS: The results showed significant anatomical differences: with the crow's foot site > forearm > calf, for skin surface hydration and corneocyte numbers; and the crow's foot site > forearm and calf, with no difference between the latter two sites, for TEWL measurements. With these techniques, seasonal differences were observed mainly in the calf, to a lesser extent in the forearm, but not in the crow's foot area. These mainly involved increases in these three parameters in July, as opposed to the other time-points. Parameters of skin surface topography however, showed no consistent seasonal pattern, but markedly higher values were observed for the forearm in comparison to the calf and crow's foot sites, which were similar. From meteorological data obtained, the average daily maximum temperature and hours of sunshine increased to peak values in July, as did the absolute humidity derived from relative humidity data. CONCLUSION: We conclude that the results are most likely to represent changes in the SC due to climatic factors, with the calf and forearm sites being most affected in comparison to the face (crow's foot wrinkle area), which seems unaffected. The reasons for this latter site remaining unchanged may be due to its greater UV exposure, sebum content, and that the use of facial cosmetics was allowed. PMID- 11428946 TI - A bioengineering study on the efficacy of a skin protectant lotion in preventing SLS-induced dermatitis. AB - BACKGROUND/AIMS: This study evaluated the efficacy of a dimethicone skin protectant lotion against sodium lauryl sulfate (SLS)-induced irritant contact dermatitis (ICD) by clinical visual grading and bioengineering techniques in 12 healthy humans. METHODS: The flexor aspects of both forearms of the subjects were used as test sites. Each test was duplicated to diminish the variations of the test sites. In a random order and a double-blind manner, two test sites were pretreated either with the testing protectant lotion or with its vehicle control prior to contact with SLS. Thirty minutes later, 0.2 ml of 0.5% SLS in a polypropylene chamber was applied to each pretreated site. One additional test site served as a positive control (without lotion), receiving the irritant only. After 24 h of exposure to the irritant, the chambers were removed. The efficacy of protective effect was determined by four parameters: visual scoring (VS), transepidermal water loss (TEWL), skin color (a* value), and cutaneous blood flow volume (BFV). All test sites were assessed with the parameters daily for 5 days. RESULTS: The VS data showed a significant decrease on the site pretreated with protectant lotion in comparison with the SLS-only treated site (P<0.01) and with the site pretreated with control vehicle (P<0.05) (overall for 5 days). TEWL value was significantly decreased in comparison with the SLS-only treated site (P=0.02 at day 2; P=0.008 at day 4; P=0.014 at day 5) and with the site pretreated with the control vehicle (P<0.05) (day 2, 4 and 5). However, the BFV and a* values did not show a statistical difference between protectant lotion, vehicle, and SLS-only treated sites. CONCLUSIONS: This study demonstrated that appropriate dimethicone skin protection products may provide certain benefits from surfactant ICD. The skin protectant lotion may be used to prevent ICD in home or work environments, where skin irritants may induce dermatitis or eczema. PMID- 11428947 TI - Electrical impedance measurements at different skin sites related to seasonal variations. AB - BACKGROUND/AIMS: In a previous study we mapped differences in electrical impedance between different anatomical locations related to age and sex. Now we employ the electrical impedance technique for studying the seasonal variations for the same skin sites. METHOD: The investigation was performed on 48 healthy volunteers divided into two groups on the basis of sex. Readings of electrical impedance were taken from ten different body locations at each of the four seasons during one year. Electrical impedance spectra were measured in the frequency range 1 kHz to 1 MHz, and four impedance indices based on the complex impedance values at 20 kHz and 500 kHz were calculated. RESULTS: The baseline values varied from one site to another and increased significantly for most sites from the winter to the summer, while the changes from October to March were negligible. Using ANOVA, the seasonal variation was found to be a more important factor than sex. CONCLUSIONS: Our results suggest that seasonal variations for different skin sites should be taken into consideration when using the electrical impedance technique. PMID- 11428948 TI - Axillary pH and influence of deodorants. AB - BACKGROUND/AIMS: In moist intertriginous regions, such as the armpit, the pH value is physiologically higher than in other skin regions. The regulation of the axillary pH-value was examined in an open study with 48 subjects in three groups with n=16 each. METHODS: In the first 10 days (run-in) the subjects received a standard treatment in the axilla with shaving, cleansing and application of a pH neutral deodorant. This was followed by a 5 day treatment period with the three test products (pH5 Eucerin(R) Deodorant Roll-on, Deodorant Balsam Spray, Deodorant Cream). The study was concluded by a wash-out period with procedures identical to the run-in phase. The pH was measured with a calibrated pH-meter. RESULTS: A significant pH reduction was shown during the treatment period when compared to the run-in phase. The Deodorant Roll-on induced a reduction of the mean pH values from 6.1 to 5.3, the Deodorant Balsam Spray from 6.5 to 5.7 and the Deodorant Cream from 6.2 to 5.3. During the wash-out period all pH values returned to baseline. CONCLUSION: All of the deodorants tested demonstrated a significant reduction in axillary pH. There is evidence that a high skin pH promotes the growth of several microorganisms that produce malodor. Therefore, the regulation of pH may contribute to the deodorant efficacy of the test products. PMID- 11428950 TI - Skin Research and Technology on electronic files, the Internet and MEDLINE/Index Medicus. PMID- 11428949 TI - Degreasing method for the seborrheic areas with respect to regaining sebum excretion rate to casual level. AB - BACKGROUND/AIMS: Sebum excreted from the seborrheic glands keeps the skin surface subtle and moist. Before determining the activity of seborrheic glands, the skin surface must be degreased to remove contamination but without provoking sebum excretion. The purpose of this study was to set up a standardised degreasing procedure for various seborrheic areas in different skin types. The method will take day-to-day variations into account with respect to the kinetics of refatting. METHODS: The Sebumeter(R) from Courage+Khazaka is used to quantify the sebum excretion. Day-to-day variations were measured on the forehead in groups of 12 volunteers on 2 consecutive days. The degreasing procedure was investigated by individual cleaning with alcohol compared to washing with a mild detergent followed by wipes with alcohol on the forehead. The degree of refatting was monitored until 3 h after defatting on seborrheic areas: the forehead, cheek, nose, chin and upper back. RESULTS: There was no statistical significant difference in the variation from day to day (P<0.05), although a change in the individual skin types was observed. An individual difference in the number of alcoholic wipes needed to degrease the forehead was seen. Washing followed by several repetitions of alcoholic wipes was not sufficient for the forehead, chin and nose (P>0.05). For the cheek and upper back, it was sufficient to wash with soap (P<0.05). The time from degreasing the skin until the sebum output reached the casual level was 2 h (P>0.05). The area under the curve (AUC) indicates that individual skin types return to casual level after defatting. CONCLUSION: There was no statistically significant day-to-day variation using the Sebumeter(R). We ended up with different degreasing procedures in different seborrheic areas. Washing gently with a detergent solution and then performing three wipes with alcohol was optimal for degreasing the forehead, nose and chin. For the cheek and upper back, it was sufficient to use a mild soap. Casual level was reached after 2 h in all seborrheic areas, irrespective of level of oiliness. PMID- 11428951 TI - Electron paramagnetic resonance: a new techniquein skin research. AB - BACKGROUND/AIM: Electron paramagnetic resonance (EPR) spectra of nitroxide spin probes have been used for studying biological membranes and chemical-membrane interaction. We have investigated the influence of surfactants on the intercellular lipid structure of cadaver stratum corneum and the possibility of EPR spectral measurements on the stripped stratum corneum utilizing cyanoacrylate resin, which might reflect the actual skin lipid conditions. CONCLUSION: EPR spectra are useful in evaluating the fluidity measurement of stratum corneum of cadaver skin and stripped stratum corneum. PMID- 11428952 TI - Electron paramagnetic resonance study utilizing stripping method on normal human stratum corneum. AB - BACKGROUND/AIM: Electron paramagnetic resonance (EPR) spectra of nitroxide spin probes are useful for studying biological membranes, and chemical-membrane interaction. The objective of this study was to assess the fluidity of stripped stratum corneum (SC) of normal human skin with EPR. METHODS: We quantified the fluidity of stripped SC (S-SC) by a single stripping with cyanoacrylate onto quartz glass, cover glass or transparent film. Spectra were compared with those of the SC of cadaver skin (C-SC). RESULTS: All S-SC spectra were similar to those of C-SC. There were no statistical differences in the order parameters S between C-SC and those on quartz cell and cover glass, but those of transparent film were significantly higher. Spectra after a month storage at room temperature at 5 degrees C and at -20 degrees C were unchanged. The order parameter S had decreased after a 3-month storage in all conditions. CONCLUSION: This method is useful for the evaluation of the fluidity measurement utilizing EPR spectra of S SC. PMID- 11428953 TI - Autofluorescence spectrum of skin: component bands and body site variations. AB - BACKGROUND/AIMS: The autofluorescence spectrum of skin consists of a number of overlapping single spectra emitted by fluorescent molecules. The overlapping makes it difficult to distinguish changes in the overall spectrum and the molecular basis for it. We applied curve fitting analysis to decompose the autofluorescence spectrum of normal human skin and studied the variation in different body sites. METHODS: Skin autofluorescence spectra were obtained in vivo from the normal skin of 17 subjects, using 5 excitation wavelengths: 340, 350, 360, 370 and 380 nm. The spectra were displayed in the wavelength range of 400 nm to 800 nm. Curve fitting was performed for each spectrum using the Levenberg-Marquardt algorithm. RESULTS: The skin autofluorescence spectra, provoked by all the excitation wavelengths, consisted of three component bands with center wavelengths at 450 nm, 520 nm and 625 nm. The 450 nm band contributed about 75% to the intensity of the overall spectrum, the 520 nm band contributed 25%, and the 625 nm band contributed about 2%. The three bands may correspond mainly to the fluorescence of nicotinamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD), and porphyrins. Significant differences were found between the spectra of forehead and buttock skin. In forehead skin, the 625 nm band contributed more to the intensity of the overall spectrum, while the 450 nm and 520 nm bands had much lower peak heights, and the 450 nm and 625 nm bands were broader. CONCLUSIONS: The autofluorescence of skin is an overlapped spectra of molecules such as NADH, FAD, and porphyrins. These molecules contribute differently to the overall spectrum in different body sites. PMID- 11428954 TI - Characterisation of gravity-induced facial skin oedema using biophysical measurement techniques. AB - BACKGROUND/AIMS: In humans, the microgravity environment can be expected to induce swelling of facial tissues and shrinking of the tissues in the lower limbs, together with a loss in body weight. To evaluate fluid shifts in skin, the head-down bed-rest model was used. The aim of the present study was to evaluate the appearance of facial oedema in subjects undergoing anti-orthostatic bed-rest at an angle of -10 degrees. METHODS: The forehead of each of four subjects was measured before and after 1, 10 and 24 h in this head-down tilt position. At these time points, interstitial fluid migration and facial oedema were assessed using a high resolution B-scan ultrasound and a device for measuring the skin's mechanical properties. RESULTS: The results obtained showed a progressive increase in dermal thickness and initial stress, and a reduction in stiffness and elasticity of the skin during the study period. CONCLUSIONS: This preliminary study has demonstrated the feasibility of the method in measuring fluid displacement and retention in the skin. Furthermore, it highlights the influence of fluids on the mechanical behaviour of the skin. These techniques could be used for studying the redistribution of liquid masses during periods spent in space. PMID- 11428955 TI - Characterization of the physical properties of the stratum corneum by a new tactile sensor. AB - BACKGROUND/AIMS: The physical properties of the stratum corneum (SC) change with its water content, which is regulated by the presence of water solutes (natural moisturizing factors) and lipids in the SC, and are considered to be responsible for the induction of desquamation, skin surface roughness, and fine wrinkles. Recently a new type of tactile sensor developed for evaluating the physical properties of objects has been introduced as a simple, non-invasive method to evaluate them; because the data obtained with this sensor have not yet been characterized in detail, we compared them with other physical parameters of the skin. METHODS: A change in resonant frequency (Deltaf ) was measured under various levels of pressure applied by a tactile sensor placed on the cheeks of 29 women. We also measured high-frequency conductance that reflects the hydration state of the skin surface, water holding capacity of the SC, transepidermal water loss (TEWL), skin flexibility, skin elasticity, amino acid composition of the SC, and desquamation index of the SC at the same time, and evaluated the correlation between Deltaf and other physical parameters. RESULTS: The correlation between Deltaf and high-frequency conductance of the SC, and that between Deltaf and the water holding capacity of the SC were substantially high. Deltaf under high pressure was more closely correlated with the acidic amino acid ratio of the SC. Deltaf also showed a correlation with desquamation index for thickness of the SC as well as with skin elasticity, independent of the applied pressure. CONCLUSIONS: Because Deltaf determined with the tactile sensor correlated with other physical parameters specific to the properties of the SC, such as high frequency conductance of the SC, its water holding capacity, ratio of acidic amino acids, and the desquamation index for thickness, the physical properties of the SC may be adequately evaluated with the measurements of Deltaf. PMID- 11428956 TI - Lesion classification using skin patterning. AB - BACKGROUND/AIMS: The observation that skin line patterning tends to be disrupted by malignant but not non-malignant skin lesions suggests that this could be used as an aid to lesion differentiation. Since recognised differentiating features can be obtained from the simply-captured white light optical image, the possibility of using such images for skin pattern disruption detection is worth exploring. METHODS: The skin pattern has been extracted from optical images by high-pass filtering and profiles of local line strength variation with the angle estimated using a new consistent high-value profiling technique. The resultant profile images have been analysed using a novel region-based agglomerative clustering technique (mRAC) and also a local variance measurement. A measure based on the relationship between the classification results and an intensity based segmentation was calculated, and this represented the disruption of the skin line patterning. RESULTS: A set of images containing a variety of histologically confirmed malignant and non-malignant lesions was analysed. The computed textural disruption figure was compared to both the histological diagnosis and to a visual estimate of patterning disruption for each image. It was demonstrated that lesion separation could be achieved by both analysis methods, with a good correlation with visual estimate of disruption and with mRAC providing the best performance. CONCLUSIONS: It was concluded that the acquisition and modelling of skin line patterning from clinical images of skin lesions had been successfully achieved and that the analysis of the resulting data provided an assessment of pattern disruption that is both consistent with visual inspection and effective in presenting information useful for discrimination between melanoma and benign naevi lesion examples. PMID- 11428957 TI - Detection of solid pigment in dermatoscopy images using texture analysis. AB - BACKGROUND/AIMS: Epiluminescence microscopy (ELM), also known as dermoscopy or dermatoscopy, is a non-invasive, in vivo technique, that permits visualization of features of pigmented melanocytic neoplasms that are not discernable by examination with the naked eye. ELM offers a completely new range of visual features. One such feature is the solid pigment, also called the blotchy pigment or dark structureless area. Our goal was to automatically detect this feature and determine whether its presence is useful in distinguishing benign from malignant pigmented lesions. METHODS: Here, a texture-based algorithm is developed for the detection of solid pigment. The factors d and a used in calculating neighboring gray level dependence matrix (NGLDM) numbers were chosen as optimum by experimentation. The algorithms are tested on a set of 37 images. A new index is presented for separation of benign and malignant lesions, based on the presence of solid pigment in the periphery. RESULTS: The NGLDM large number emphasis N2 was satisfactory for the detection of the solid pigment. Nine lesions had solid pigment detected, and among our 37 lesions, no melanoma lacked solid pigment. The index for separation of benign and malignant lesions was applied to the nine lesions. We were able to separate the benign lesions with solid pigment from the malignant lesions with the exception of only one lesion, a Spitz nevus that mimicked a malignant melanoma. CONCLUSION: Texture methods may be useful in detecting important dermatoscopy features in digitized images and a new index may be useful in separating benign from malignant lesions. Testing on a larger set of lesions is needed before further conclusions can be made. PMID- 11428958 TI - Tonality of suntan vs sunless tanning with dihydroxyacetone. AB - BACKGROUND/AIMS: Although there is an increasing awareness of the detrimental effects of solar irradiation on skin, a tanned look is still in fashion. To achieve the tanned look without sun exposure various sunless tanning formulations have become available. Most of these contain dihydroxyacetone (DHA) which binds to the proteins of the stratum cornium and imparts a brown color to skin. This color is similar to a suntan but can be somewhat more yellow, making it appear unnatural. The aim of this study was to determine a quantitative method to define a "natural" tan and to study methods to improve the tonality of sunless tanning on skin. METHODS: Tonality of suntan was determined as the marker for a "natural" tan. In order to achieve this, human volunteers were exposed to the sun and the change in skin color was observed after 3 days. Color measurements obtained with the Minolta Chromameter were plotted in two standard graphs labeled as the "natural universe of tan", which depicted a balance between Chroma and change in reflectance, while the "natural universe of color" determined the balance between changes in yellow and red components of the suntan. Change in skin color after treatment with DHA was then inserted in these graphs to observe how "natural" the tonality of sunless tan appeared relative to suntan. RESULTS: Results show a good balance between Chroma and change in reflectance for suntanned skin in the "natural universe of tan". Conversely, the "natural universe of color" exhibited a good balance between increase in yellow and red components of suntan. Sunless tan data inserted in these graphs showed that for several subjects with skin types I-II the tonality is not within the realm of "natural" but is unnaturally yellow. Addition of antioxidants, especially caffeic acid phenethyl ester (CAPE) in DHA formulation significantly shifted the tonality towards the center of the "natural universe of color". CONCLUSIONS: The "natural universes of tan and color" exhibit a simple and quantitative assessment of tonality of suntan and sunless tan. Although skin tonality from DHA-induced sunless tan can often lie outside the realm of the "natural" tan, it is possible to improve this tonality to a more "natural" look by addition of strong antioxidants in the DHA formulations. PMID- 11428959 TI - Study of three complementary techniques for measuring cutaneous hydration in vivo in human subjects: NMR spectroscopy, transient thermal transfer and corneometry - application to xerotic skin and cosmetics. AB - BACKGROUND/AIMS: The aim of this study was to determine the capability and the analytical quality of three different in vivo, non-invasive, quantitative methods for measuring skin hydration: two innovative methods that have been used for more than eight years - nuclear magnetic resonance spectroscopy (NMR-S) and transient thermal transfer (TTT) - and the more widely used and conventional corneometry. METHODS: The work presented evaluated the capability and precision, as well as cutaneous exploration depths, of the three methods. Experiments were carried out in vivo following the hydration, in kinetic terms, induced by topic application of reference moisturizing products. Spatio-temporal efficacy of a lipolotion was also studied by the TTT METHOD: Cases of xerotic skin were studied with TTT and corneometry. RESULTS: The results obtained showed better repeatability and reproducibility with the TTT and NMR-S methods than with corneometry. NMR-S is one of the only direct hydration measurement methods. It measures skin hydration down to the outer dermis with high precision. It is indicated for products having an action down to the deep cutaneous layers. By changing thermal power parameters, the TTT method can determine hydration to the outer, middle or deep epidermal layers. It is, therefore, possible to track the penetration of products in various layers of the epidermis. The small size of the probe enables the hydration measurement of skin sites (lips, eyelids) that were not, up to now, measurable with the two other methods. Corneometric investigations are restricted to the surface of the horny layer; measurements are easy and rapid but influenced by the composition of products applied to the skin and their phases: aqueous, oily or ionic. The xerotic skin study highlights the importance of exploration in different layers of the epidermis, as dehydration concerns not only the upper layers of the epidermis but also the medial and deep layers. With the TTT method, it has been possible to highlight the penetration dynamics of a lipolotion with, initially, an increase in the hydration in the outer epidermis, followed 3 h later by a transfer from the outer to the middle epidermis. CONCLUSION: NMR-S, TTT and corneometry represent three possible ways to assess skin hydration. Because they explore different cutaneous depths, they are more complementary than competitive. Transient thermal transfer, although a semi-direct method, is a precise, informative, and innovative solution to evaluate skin hydration at different epidermal depths and sites. PMID- 11428960 TI - In vivo model of the mechanical properties of the human skin under suction. AB - BACKGROUND/AIMS: A new method for the in vivo characterization of the mechanical properties of skin has been developed. This comprises a suction chamber and an ultrasound device to measure both the vertical displacement of the skin's surface, and the skin's thickness. METHODS: A mathematical model of the mechanical behaviour of a taught elastic membrane is used to obtain a set of parameters intrinsic to the skin, such as Young's modulus (E) and the initial stress (sigma0), which reflect the stiffness and the natural tension of the skin, respectively. We also calculated an index of non-elasticity of the skin (unrestored energy ratio, UER), which takes into account the volume of tissue mobilized. It determines a ratio between the energy input to the skin and the energy it dissipates. These parameters were evaluated from the volar forearm of 10 normal male volunteers. RESULTS: The results were: 129+/-88 kPa for E, 13.5+/ 5 kPa for sigma0, and 0.42+/-0.04 for UER; with reproducibilities of 9.5%, 12.4% and 6.4%, respectively. CONCLUSIONS: This new suction device was found useful for the study of the behaviour of the skin, and the device may be used for the evaluation of certain skin diseases and their therapy. PMID- 11428961 TI - Quantitative assessment of properties of make-up products by video imaging: application to lipsticks. AB - BACKGROUND/AIMS: The different properties and visual effects of lipstick have been studied by image analysis directly on volunteers. METHODS: After controlling the volunteer's position mechanically using an ophthalmic table and visually using an acquirement mask, which is an indicator of luminance and guide marks, we carried out video colour images of the make-up area. From these images, we quantified the colour, gloss, covering power, long-lasting effect and streakiness, using computer science programs. RESULTS/CONCLUSION: Quantitative colorimetric assessment requires the transformation of the RGB components obtained by a video colour camera into CIELAB colorimetric space. The expression of each coordinate of the L*a*b* space according to R,G,B was carried out by a statistical method of polynomial approximations. A study, using 24 colour images extracted from a Pantone(R) palette, showed a very good correlation with a Minolta Colorimeter(R) CR 300. The colour assessment on volunteers required a segmentation method by maximizing the entropy. The aim was to separate the colour information sent back by the skin to the make-up area. It was very useful to precisely delimit the contour between the skin and the product in the case of almost identical colours and to evaluate the streakiness. From this colour segmentation, an algorithm was studied to search for the shades most represented in the overall colour of the make-up area. The capacity to replicate what the consumer perceives of the make-up product, to carry out studies without having any contact with the skin surface, and the constant improvement of software and video acquirement systems all make video imaging a very useful tool in the quantitative assessment of the properties and visual effects of a make-up product. PMID- 11428962 TI - Skin color measurements: comparison between three instruments: the Chromameter(R), the DermaSpectrometer(R) and the Mexameter(R). AB - BACKGROUND/AIMS: Two types of skin reflectance instruments are available nowadays for the determination of skin color: a tristumulus colorimeter (Chromameter from Minolta) using the CIE L*a*b* color system and the narrow-band simple reflectance meters (DermaSpectrometer from Cortex and Mexameter from Courage-Khazaka) using the erythema/melanin indices. The purpose of this study was to compare the capabilities of the three instruments (sensitivity, repeatability and correlation) in vitro and in vivo. METHODS: Comparative color measurements were carried out first in vitro on standardized color charts and subsequently in vivo on different skin areas in human volunteers. Skin color changes induced by various physico-chemical treatments were also quantitatively evaluated with the three instruments. RESULTS: The in vitro and in vivo repeatabilty as well as the sensitivity of the three instruments are rather good. Erythema and skin blanching could be readily quantified by the increase of the a* parameter and of the erythema indices of the simple reflectance meters. Natural UV tanning and artificial chemical tanning could be measured by the decrease of L* and increase of b* and of the melanin indices. CONCLUSION: The Chromameter and the two narrow band reflectance instruments were able to characterize skin color and to quantify small skin color changes. Moderate to high significant linear correlations could be established between the CIE L*a*b* color parameters and the erythema/melanin indices. PMID- 11428963 TI - Use of Cutometer to assess epidermal hydration. AB - BACKGROUND/AIMS: The Cutometer equipped with a 2-mm diameter suction probe is a device suitable for assessment of epidermal mechanics. The objectives of this study were to determine the sensitivity of Cutometer parameters to epidermal hydration, to evaluate the correlation between skin mechanical parameters and skin capacitance, and to study the role of pretension of the skin. METHODS: In the first study, skin capacitance (Corneometer) and 10 mechanical parameters were determined before and 60 min after application of five different moisturizers on the volar forearm. In the second study, measurements of epidermal mechanics were made without and with 0.1 s presuction of the skin, before and 120 min after application of petrolatum and glycerin. RESULTS: Hydration of the skin significantly increased the values of capacitance and all rheological parameters. Delayed distension (Uv) and viscoelastic-to-elastic ratio (Uv/Ue) were the most sensitive Cutometer parameters. Significant correlations between skin capacitance and mechanical parameters were not found. Pretension of the skin did not significantly influence the values of skin mechanical parameters. CONCLUSIONS: The non-invasive measurements of skin elasticity are appropriate for an objective and quantitative evaluation of the complex effect of different dermatological and cosmetic products on skin mechanics and hydration. PMID- 11428964 TI - Comparison of surface free energy between reconstructed human epidermis and in situ human skin. AB - BACKGROUND/AIMS: The objectives of this study were to characterise the Episkin(R) model of reconstructed epidermis histologically and energetic, and to compare the results with the data obtained by Mavon (9, 10) on different areas of skin on the human body and on different skin types. METHODS: Histologically, the two skin systems studied were stained using Goldner's trichome METHOD: Surface free energy was determined by the measurement of contact angles, using the posed drop METHOD: RESULTS: Histologically, the two skin systems demonstrated the same cellular organisation in stratified layers. With regard to surface free energy, Episkin, skin from the forehead, normal and hyper-seborrheic skins are hydrophilic surfaces, while skin from the forearm or of an oligo-seborrheic type tends to be hydrophobic. Episkin and skin from the forehead exhibit similar surface free energy profiles. It is impossible to state whether Episkin has characteristics closer to those of normal or hyper-seborrheic skin, but they differ significantly from those of oligo-seborrheic skin. CONCLUSION: The results obtained during this study justify the use of Episkin in a range of tests where the use of in situ human skin raises problems. PMID- 11428965 TI - Evaluation of an experimental patch test model for the detection of irritant skin reactions to moisturisers. AB - BACKGROUND/AIMS: Moisturisers are used daily by a large number of people to prevent dryness of the skin. Irritant skin reactions to moisturisers are, however, known to occur. In order to prevent such irritant reactions reliable test methods for irritancy testing of moisturisers are needed. This study was undertaken to evaluate a non-invasive patch test model for the detection of irritant skin reactions to moisturisers. METHODS: Twenty healthy volunteers were patch tested with three different moisturisers: empty chamber, sodium lauryl sulphate and a moisturizer known to be non-irritating. Skin reactions were evaluated by visual scoring, measurement of transepidermal water loss (TEWL) by an Evaporimeter, blood flow by laser Doppler flowmetry and electrical capacitance by a Corneometer. RESULTS: A statistically significant increase in blood flow was found 48 h after application of one of the moisturisers tested, indicating an irritant effect of the product. A statistically significant decrease in skin hydration was found for the same moisturiser after 48 h. No statistically significant differences between the moisturisers were found by visual scoring. None of the products tested had any negative effect on the skin barrier function. CONCLUSION: The non-invasive patch test model was found useful for detecting irritant skin reactions to moisturisers. PMID- 11428967 TI - Opportunistic infections after blood and marrow transplantation. AB - Opportunistic infections are major causes of morbidity and mortality following bone marrow transplantation. Technological advances in stem cell procurement, the introduction of hematologic growth factors to speed engraftment, the development of new immunosuppressive regimens to control graft-versus-host disease (GVHD), the development of technology to perform graft engineering with removal of T lymphocytes in toto or subpopulations of T lymphocytes, the use of molecular techniques to optimize donor and recipient matching, advances in blood banking, and development of international donor registries, are among the various factors that have led to tremendous changes in transplant practices. Because of such changes in transplant practices, along with the advent of new antimicrobial agents, and development of infection control measures affecting pathogen exposure, alterations in the interplay between host and potential pathogens have occurred. Shifts in the incidence and types of opportunistic pathogens are taking place. Several historically important infectious syndromes are today well controlled; others have diminished in importance early after transplant but are more problematic at a later time; new emerging pathogens are being recognized due to selection pressures from antimicrobial usage and new hosts, such as recipients of alternate donor allogeneic transplant procedures, with even more profound and prolonged immune suppression. Such shifts and new syndromes pose continuing new challenges to the transplant clinician. PMID- 11428968 TI - Infection in solid organ transplantation. AB - Infection in the organ transplant recipient is a major determinant of the outcome of transplantation. The attributable mortality from invasive fungal disease and cytomegalovirus disease has been 50 and 30%, respectively. Determinants of infection include epidemiologic exposures to environmental pathogens or exposure to latent infection in the donated organ or recipient, and changes in the net state of immunosuppression. For the past two decades significant improvements have been made in the recognition, prevention and treatment of infections, such as cytomegalovirus and P. carinii. Future research will need to focus on treatment of invasive mold infections, and the management of hepatitis B and C in liver transplantation. In addition, improvement in the prevention of the post transplant lymphoproliferative disease syndrome will be necessary. PMID- 11428969 TI - The therapeutic prescription for the organ transplant recipient: the linkage of immunosuppression and antimicrobial strategies. AB - Infection and rejection, the two major barriers to successful organ transplantation, are closely linked, with immunosuppressive therapy being central to the pathogenesis of both. After almost two decades when azathioprine and prednisone, supplemented by antilymphocyte antibody therapy, were the cornerstones of post-transplant immunosuppressive programs, there has been a major increase in the therapeutic armamentarium available to treat rejection: cyclosporine, tacrolimus, mycophenolate mofetil, rapamycin, and antibodies directed against the interleukin-2 receptor. These agents are potent inhibitors of microbial specific T cell function, thus potentiating opportunistic infection with herpes group viruses, fungal and mycobacterial species, Strongyloides stercoralis, and a variety of intracellular pathogens. The mechanisms by which each of these drugs exerts its effects are an important determinant of the antimicrobial strategies that will be necessary to combat infection. Indeed, strategies to limit these infections are being linked to the nature of the immunosuppressive therapy required in a particular patient. Thus, the therapeutic prescription for the transplant patient is said to have two components: an immunosuppressive component to prevent and treat rejection, and an antimicrobial one to make it safe. In addition to using antimicrobial agents therapeutically, in the transplant patient prevention is stressed in which antibiotics are deployed prophylactically or preemptively. PMID- 11428970 TI - Preventing opportunistic infections in bone marrow transplant recipients. AB - In 1996, a Center for Disease Control and Prevention (CDC)-sponsored working group began developing guidelines for preventing opportunistic infections (OIs) in bone marrow transplant (BMT) recipients. The purposes of the guidelines are to: a) summarize current data regarding the epidemiology of OIs in BMT recipients; b) produce an evidence-based statement of recommended strategies for preventing OIs in BMT recipients; c) decrease the incidence, morbidity, and mortality of OIs in BMT recipients; and d) define directions for future OI prevention research. Each recommendation is given two ratings: one indicating the strength of the recommendation, and another indicating the strength of evidence supporting the recommendation. The target audience for the guidelines includes transplant and infectious disease physicians and BMT unit and clinic staff. The BMT OI guidelines include sections on viral, bacterial, fungal, protozoal, and helminth infections, immunization, infection control, and blood and stem cell safety. The disease-specific sections address preventing exposure and disease among both adult and pediatric recipients of allogeneic and autologous BMTs. The immunization section addresses: a) immunization of BMT recipients, their household contacts, and health care workers; b) travel immunizations for BMT recipients; and c) passive immunization with immune globulin products. The infection control sections address room ventilation, isolation and barrier precautions, and prevention of nosocomial and other infections (e.g. infections acquired from visitors, plants, food, pets, construction sites, etc.). The blood safety section contains recommendations on preventing transmission of infections to BMT recipients from infected donated cells. After the guidelines are made available for public comment, they will be finalized and published in the Morbidity and Mortality Weekly Report and placed on the CDC web site. PMID- 11428971 TI - Prevention of infections in solid organ transplant recipients. AB - Solid organ transplantation has become an important therapeutic option for multiple chronic diseases. With the advent of newer and potent immunosuppressive regimens, graft survival has improved, but at the expense of an increased risk for the development of infections secondary to bacterial, fungal, viral and parasitic pathogens. Prevention of such infectious complications with effective, well tolerated, and cost-effective anti-microbials would be ideal to improve the outcome of transplanted patients. However, the emergence of multi-drug resistant pathogens, medication toxicity and drug-drug interactions need to be carefully evaluated. This review summarizes the most relevant data pertaining to our current understanding of infection prevention for solid organ transplant recipients. Specific recommendations are given for the prevention of each group of microorganisms and types of solid organ transplant. PMID- 11428972 TI - Immunosuppression without immunosuppression? How to be a tolerant individual in a dangerous world. AB - The field of transplantation has developed based on two principles: allografts are rejected because they express foreign antigens, and the immune system must be suppressed to prevent rejection. Recently, in vitro and in vivo experimental evidence has accumulated that calls both of these beliefs into question. This article reviews an alternative approach to transplantation that focuses on tissue injury as the instigator of graft rejection and employs physiological mechanisms of tolerance to avoid graft loss. Methods that allow for defense against infectious microbes while at the same time allowing for graft survival are proposed. In particular, the rationale behind the use of anti-CD154 antibody treatment is highlighted. A model is introduced that takes into consideration the experimental successes seen with anti-CD154 therapies. PMID- 11428973 TI - Abdominal complaints in a long-term renal transplant patient with a failing allograft. PMID- 11428974 TI - Tuberculosis isolated to the renal allograft. AB - Immunosuppressed patients after renal transplantation are at increased risk of developing infective complications, including tuberculosis. We describe four renal transplant patients who presented with tuberculosis isolated to the renal allograft. The three patients with adequate follow-up presented with a febrile illness 12-26 months after the transplant. The mycobacterial infection was found in the graft nephrectomy specimen in three patients. One patient with renal allograft dysfunction had a percutaneous biopsy that showed tuberculosis. The latter patient recovered renal allograft function after anti-tuberculous therapy. None of the patients had evidence of tuberculosis elsewhere. We conclude that tuberculosis isolated to the renal allograft, especially in an endemic area, may be the cause of renal dysfunction, and appropriate therapy could lead to salvage of the graft. PMID- 11428975 TI - Cytomegalovirus and transplantation: yesterday, today, and tomorrow. PMID- 11428976 TI - High-dose acyclovir prophylaxis reduces cytomegalovirus disease in liver transplant patients. AB - Cytomegalovirus (CMV) is still a major pathogen in liver transplantation (LTX). The clinical efficacy of prophylactic high-dose acyclovir therapy (800 mg qid) was assessed for the prevention of CMV infection and disease in liver recipients. Fifty-five patients were enrolled in a prospective, randomised, double-blind and placebo-controlled trial; 28 on acyclovir vs. 27 on placebo. The therapy was given for 12 weeks. The patients were followed for 24 weeks. CMV infection was diagnosed in 60% (16 on acyclovir, 17 on placebo) and CMV disease developed in 38% (7 on acyclovir, 14 on placebo) of the patients. The total mortality was 27% (6 on acyclovir, 10 on placebo). Acyclovir delayed 32% of the CMV infections and prevented 59% of the CMV disease cases which occurred in the placebo cohort. The time to CMV disease was significantly prolonged in patients on acyclovir as compared to patients on placebo (P=0.013). Adverse events included neurotoxicity which occurred in 5 cases in the acyclovir, but none in the placebo arm, and nephrotoxicity which was detected in 6 patients in the acyclovir and 5 in the placebo arm, respectively. We conclude that acyclovir prophylaxis significantly reduced the incidence of CMV disease, and delayed the onset of CMV infection in liver transplant patients. PMID- 11428977 TI - Tuberculosis in renal transplant recipients. AB - Tuberculosis (TB) has been described in kidney transplant recipients as an infection with predominantly pulmonary involvement. We report the impact of TB in kidney transplantation. Clinical records of adult kidney recipients, transplanted between 1 January 1986 and 31 December 1995 were analyzed for sex, age, graft origin, immunosuppressive therapy, TB sites, diagnostic methods and concomitant infections. Annual incidence, mean time of onset, relation to rejection treatment, tuberculin skin test (PPD) and outcome were analyzed. Patients with a history of TB or graft loss in the first month were excluded. TB was diagnosed in 14 of 384 (3.64%). Mean age at transplantation was 35 years. Twelve of these received the graft from a living donor. All had triple immunosuppression with cyclosporine. Ten had pulmonary TB, three extrapulmonary infection and one disseminated disease. In 13 cases an invasive diagnostic procedure was performed. Mycobacterium tuberculosis cultures were positive in all cases; microscopy revealed acid-fast bacilli (AFB) in 6, and adenosine deaminase was elevated in CSF and pleural effusion in 2. Annual incidence varied from 0% to 3.1%. At the time of TB presentation 8 patients had other concomitant infections (cytomegalovirus, nocardia, Pneumocystis carinii, disseminated herpes simplex virus). Median time of onset was 13 months. Diagnostic results became available post-mortem in 2 cases, and one had TB in a failing allograft. TB was treated with 4 drugs including rifampin in 10 patients. Cyclosporine was discontinued in one, lowered in one and increased in 8. During treatment 5 patients had rejection episodes. At 1 year, graft survival was 72.7% and patient survival 90.9%. TB was more prevalent when recipient and donor were both PPD positive. In summary: although TB is a growing threat in the transplant setting, early and aggressive diagnosis with meticulous monitoring of immunosuppression allows a successful outcome for both patient and graft. Optimal prophylaxis guidelines have yet to be completely defined. PMID- 11428978 TI - Antiviral drug resistance in human cytomegalovirus. AB - Drug-resistant cytomegalovirus (CMV) should be considered when viral shedding persists after several weeks of therapy. The problem is most likely to arise in the setting of a severely immunosuppressed host with continuing or relapsing disease. Not all treatment failure can be attributed to drug resistance. The testing of CMV isolates for drug resistance in cell culture is time-consuming and labor-intensive, but recent advances in understanding of the genetics of resistance have resulted in rapid genotypic assays for specific mutations in the viral UL97 phosphotransferase or UL54 DNA polymerase genes that can predict resistance and cross-resistance to specific drugs. This information may help in the selection of alternative therapy. PMID- 11428979 TI - Cytomegalovirus infection and cardiac allograft vasculopathy. AB - There is a wealth of clinical and experimental evidence indicating the interaction of cytomegalovirus (CMV) infection and rejection in cardiac and other solid organ allografts. A plausible explanation for this association comes from data showing that therapy with biologicals, sepsis, and rejection, all lead to the release of TNF-alpha which, upon binding to its receptor, activates NF-kB. TNF-alpha is also able to stimulate the activity of the CMV-IE enhancer/promoter region. CMV infection of several cell lines leads to NF-kB activation. NF-kB binding sites are present in regulatory regions of various cellular and viral genes, including the IE enhancer region of CMV. In a reciprocal situation, CMV infection, most likely via gamma-interferon, leads to upregulation of MHC antigens in the transplant and, thereby, to increased transplant immunogenicity. Thus, a vicious circle is induced. We have investigated in detail the pathobiology of CMV and allograft vasculopathy (chronic rejection) in experimental animals, using aortic and cardiac allografts as well as a trachea model. The results may be summarized as follows: Infection of the recipient with rat CMV results in an early inflammatory response in the aortic and cardiac allograft vascular adventitia and intima (endothelialitis) and in the airway wall of tracheal allografts. This early inflammatory response leads to enhanced intimal thickness in aortic and cardiac allografts and enhanced luminal occlusion of tracheal allografts. Timewise, this coincides with early activation of intragraft inflammatory leukocytes and increased mRNA of various growth factors and cytokines. When the recipients receive gancyclovir, the enhanced intimal response in aortic and cardiac allografts and luminal occlusion in tracheal allografts is entirely abolished. Gancyclovir treatment dramatically reduces the inflammatory response in the allograft, and thereby growth factor synthesis in response to injury. However, gancyclovir does not prevent the expression of IE antigen of CMV, suggested to inactivate tumor suppressor protein p53 predisposing smooth muscle cells to increased growth. Taken together, the effect of CMV infection on cardiac allograft dysfunction is bidirectional and biphasic. The bidirectional nature emerges from the observations that acute CMV infection may accelerate acute rejection, and, on the other hand, acute alloimmune response associated cytokine response may activate latent CMV infection. The biphasic effect of CMV on allograft dysfunction refers to its early and late detrimental effects, i.e. during the time of acute and chronic rejection. These two effects of CMV on allograft dysfunction emphasize the need for precise diagnosis of CMV infection in transplant recipients and pre-emptive or prophylactic anti-viral therapy. The benefits of this strategy may not be evident during the early post transplant period, but 5-10 years after transplantation they manifest as better graft survival. PMID- 11428980 TI - Molecular pathogenesis of human cytomegalovirus infection. AB - Despite progress in diagnosis and treatment of human cytomegalovirus (CMV) infection, we do not understand why, in hosts with comparable levels of immunosuppression, some CMV infections result in symptomatic CMV disease while others are limited to asymptomatic virus shedding with no discernible clinical consequences. CMV viral detection and quantification are useful for identifying those at highest risk, but do not consistently predict clinical outcome. Factors such as host genotype and immune response are active areas of research. However, the importance of CMV strain variability, recognized since 1976, is now receiving attention. Advances in technology that allow the rapid sequencing of viral DNA for purposes of strain characterization have fueled the renewed interest. The focus of this review will be to summarize our evolving knowledge of CMV strain variability and to document where possible a potential relationship to strain virulence. Studies with the UL55 (gB) envelope glycoprotein will be emphasized because of the ability to clearly identify naturally occurring variants, as well as the increasing number of reports that there are differences in biological activities that may contribute to virulence. PMID- 11428981 TI - Reactivation of chronic Chagas' disease following allogeneic bone marrow transplantation and successful pre-emptive therapy with benznidazole. AB - This report shows the early detection of reactivation of chronic Chagas' disease (CCd) in a 27-year-old man with chronic myelogenous leukemia undergoing allogeneic bone marrow transplantation (ABMT). Pre-emptive therapy with benznidazole during a period of 7 weeks led to a rapid recovery of the patient, who remains free of parasitemia 2 years after the bone marrow transplantation. PMID- 11428982 TI - Infective endocarditis in renal transplant recipients. AB - Because of the increasing number of renal transplantations performed and the rarity of reported cases of infective endocarditis in these patients, we studied the clinical characteristics of this infection in this population. We report on two cases from our experience and review reported cases of infective endocarditis in renal transplant recipients retrieved from the MEDLINE system. In addition, we reviewed a large series of infective endocarditis looking for patients with renal transplants. In addition to our 2 cases, 12 previously reported cases were found. The mean time from transplantation to diagnosis of infective endocarditis was 3.5 years (range 2 months to 15 years). Causative organisms included fungi, Staphylococcus aureus (3 cases each), Corynebacterium sp. (2 cases), Streptococcus viridans, VRE, Brucella sp., Clostridium sp., Nocardia sp. and Erysipelothrix sp. (one case each). Skin manifestations of endocarditis and/or splenomegaly were not reported in these patients. Septic emboli and mycotic aneurysms were relatively common. The overall mortality rate was 50% (7 of 14 patients died). Infective endocarditis seems to be rare in renal transplant recipients. The few reported cases are characterized by unusual causative micro organisms and atypical clinical presentation. Further studies are needed to delineate the magnitude and scope of this association. PMID- 11428984 TI - Low HLA-DR expression on peripheral blood monocytes predicts bacterial sepsis after liver transplantation: relation with prednisolone intake. AB - Bacterial sepsis remains a frequent complication after liver transplantation. We previously reported the results of a pilot study that suggested that low expression of HLA-DR on monocytes is a predictive marker for the occurrence of sepsis. We have studied the value of this marker in an additional cohort of patients, and have analyzed the relation of HLA-DR expression with the use of immunosuppressive agents. 20 adult liver transplantation patients were prospectively monitored during the first 4 weeks after transplantation. All were treated according to standard protocols. The percentage of monocytes expressing HLA-DR was measured by flow cytometry. In addition, the effects of incubation of monocytes with prednisolone in vitro on the expression of HLA-DR was determined in 7 healthy volunteers. Seven patients developed bacterial sepsis after a median 15 (range 10-20) days after transplantation. HLA-DR expression was significantly lower in these patients on days 7, 14, 21, and 28 after transplantation compared with non-septic patients. The percentage of HLA-DR positive monocytes was 30% or less, 3 (1-8) days before onset of sepsis. On day 7 after transplantation, HLA-DR expression on 50% or less of monocytes had a positive predictive value for sepsis of 71%, whereas the negative predictive value was 85%. Patients who developed sepsis received significantly more prednisolone. Incubation with prednisolone in vitro lowered the expression of HLA-DR in a dose-dependent manner. We conclude that low HLA-DR expression on monocytes is a marker for a high risk of subsequent sepsis in liver transplantation patients. This high risk may be (at least partly) related to the dose of prednisolone. PMID- 11428985 TI - Neutrophil respiratory burst following liver transplantation: in vitro effects of granulocyte colony-stimulating factor. AB - Early postoperative infections and septic complications are predominant causes of morbidity and mortality in patients following orthotopic liver transplantation (OLTx). Prophylactic granulocyte colony-stimulating factor (G-CSF) administration after OLTx was found to decrease the number of sepsis episodes and sepsis-related mortality. Since polymorphonuclear neutrophils (PMNs) are one of the major determinants of antimicrobial defense, alteration of their functions may influence the development of sepsis in these patients. Therefore, we investigated in vitro whether or not priming with G-CSF affects the neutrophils' respiratory burst (RB) in immunosuppressed liver-transplanted patients. Venous blood was drawn from liver allograft recipients (n=12) between the 5th and 15th day postoperatively. Patients without clinical signs of infection or rejection were included in this study. Leukocytes were obtained as supernatant following sedimentation and incubated with 1000 IE ml-1 G-CSF. The RB was measured by the intracellular oxidation of non-fluorescent dihydrorhodamine to the fluorescent rhodamine by flow cytometry. The results were expressed as a percentage of increasing stimulation compared to the control responses, which are made up of the percentage of cells with RB reaction after stimulation with phorbol ester (PMA), bacteria (E. coli), or the combination of a cytokine (TNF-alpha) and a bacterial peptide (FMLP) in the absence of G-CSF. In vitro priming with G-CSF resulted in significantly increased activity of the RB after PMA (from 71.7% to 85.6%) and TNF-alpha/FMLP (from 58.4% to 72.7%) stimulation. These data demonstrate that G-CSF in vitro augments the RB of PMNs, thereby suggesting a possible therapeutic role for G-CSF as immunomodulating agent during bacterial and fungal infections following OLTx. PMID- 11428986 TI - Mechanisms of human cytomegalovirus (HCMV) (re)activation and its impact on organ transplant patients. AB - Human cytomegalovirus (HCMV) infection plays an important role in transplant patients. Its impact is both direct and indirect. This review focuses on new aspects of HCMV (re)activation and HCMV related pathology, particularly HCMV associated renal allograft injury. During the last two years we have learned that HCMV is more frequently (re)activated, even in healthy people, than previously expected. Inflammatory as well as stress mediators and some drugs may (re)activate the virus by using distinct intracellular pathways. Commonly, HCMV (re)activation is accompanied by HCMV antigenemia/DNAemia, suggesting that precursor cells in the bone marrow play an important role as a reservoir of latent virus. However, local HCMV (re)activation (colon, lung) without detection of active HCMV infection in the peripheral blood is possible. In healthy people a sufficient type 1 T-cell response controls the active HCMV infection, while in patients with severe immune deficiency (AIDS, high-dose immunosuppression) the virus can spread in an uncontrolled fashion and induce 'classic' HCMV disease. In patients with moderate immune deficiency (e.g. long-term transplant patients on low-dose immunosuppression) virus spreading is controlled but the elimination of cells harboring the active virus may be insufficient. The resulting persistent HCMV antigenemia may induce chronic inflammatory processes leading to tissue injury, particularly in the allograft. Therefore, antiviral therapy may be useful in patients suffering from graft deterioration with otherwise clinically symptomless HCMV infection. HCMV-related immune deficiency with an increased risk of developing bacterial/fungal superinfections is frequently seen in patients with symptomatic HCMV disease but not in asymptomatic CMV antigenemia. The risk of developing superinfections can be predicted by flow-cytometric monitoring of peripheral blood monocytes. PMID- 11428987 TI - Current antiviral strategies for controlling cytomegalovirus in hematopoietic stem cell transplant recipients: prevention and therapy. AB - Cytomegalovirus (CMV) remains a cause of significant morbidity and mortality after hematopoietic stem cell transplantation (HSCT). Ganciclovir prophylaxis, or preemptive treatment based on detection of antigenemia or CMV DNA by PCR, effectively prevents CMV disease during the first 100 days after transplant in allograft recipients. In recipients of T-cell depleted transplant or if severe acute graft-versus-host disease is present, ganciclovir prophylaxis or preemptive treatment should be started with an induction course of ganciclovir (5 mg/kg BID) and given at least 5 days per week and continued until day 100 after transplant. Although prevention of CMV disease before day 100 is highly effective, there is a continued risk of late-onset CMV disease after day 100. In CMV-seropositive recipients, the incidence of late CMV disease may be as high as 17%. Strategies to prevent late CMV infection and disease are needed. In seronegative recipients, seronegative or leukocyte-reduced blood products are effective in preventing acquisition of CMV through blood products. Controversy exists about the optimal strategy of preventing CMV disease in seropositive autologous HSCT recipients. The outcome of CMV pneumonia remains poor despite treatment with ganciclovir in combination with CMV hyperimmune globulin or intravenous immunoglobulin. Owing to continued clinical significance of CMV in the HSCT setting, new and more effective anti-CMV drugs with improved pharmacokinetic properties are urgently needed. PMID- 11428988 TI - Diagnostic approaches to cytomegalovirus infection in bone marrow and organ transplantation. AB - Cytomegalovirus (CMV) continues to be a clinical problem, impairing the overall success rate of transplantation, either through direct involvement of a variety of end-organs or by inducing indirect effects such as graft rejection. We review here how the virus manages to evade host immune responses and replicate extensively in allograft recipients. Recent studies show that the quantity of CMV (viral load) is related directly to the development of CMV disease. We review how clinically significant levels of CMV viral load can be defined and summarize the results of studies showing that a high CMV viral load is the major determinant of CMV disease, explaining the previously reported risk factors of pre-transplant serostatus and the post-transplant detection of CMV viremia.Note PMID- 11428989 TI - Prevention and treatment of cytomegalovirus infection in organ transplant recipients. AB - Cytomegalovirus (CMV) is the most common viral pathogen in organ transplant recipients. The patients at highest risk of developing CMV disease are seronegative recipients of seropositive donors, and seropositive recipients who receive antilymphocyte agents such as OKT3 and antithymocyte globulin (ATG) for induction or for rejection. There have been many trials of CMV prevention, but they are difficult to compare with one another because of variability in definitions and end points. Two modalities that have been used to prevent CMV disease are prophylaxis and preemptive therapy. In prophylaxis all patients are given an antiviral agent in order to prevent CMV disease, while in preemptive therapy (also called targeted prophylaxis) only patients who are identified as 'high risk' are selected for treatment. Selected trials of prophylaxis and preemptive therapy in solid-organ recipients are reviewed. The factors to be considered in using one modality or the other are side effects from antivirals, cost of monitoring and antivirals, efficacy of the two modalities, and potential emergence of drug resistance. Sensitive tests that have been used for early diagnosis and monitoring of CMV are antigenemia and the polymerase chain reaction (PCR). Antigen pp65 is a lower matrix protein and can be detected in peripheral blood leukocytes. The sensitivity and specificity are high and vary from 89% to 100% and 92% to 96%, respectively. Currently, many authors believe that the antigenemia test is more useful than the PCR test. The antigenemia test is useful for viral monitoring as a guide for preemptive therapy after organ transplantation. Persistence of high counts of antigenemia may indicate inadequate antiviral therapy or emergence of resistance. Recurrence of positive antigenemia after treatment of CMV disease can be a sign of relapse. Transplant patients who develop resistance to antiviral drugs are usually seronegative recipients who receive an organ from a seropositive donor and have several courses of antivirals for CMV disease. Ganciclovir is the most frequent antiviral agent used in transplant recipients and is usually well tolerated. Resistance to ganciclovir may occur and is usually secondary to virus mutation in the UL97 gene. The availability of sensitive diagnostic tests such as pp65 antigenemia has made the early diagnosis of CMV possible in organ transplant recipients. CMV is being treated much earlier now, and progression to disseminated disease is uncommon. Prudent use of antiviral drugs will hopefully limit the problem of drug resistance. PMID- 11428990 TI - The biology of Epstein-Barr virus in post-transplant lymphoproliferative disease. AB - Post-transplant lymphoproliferative disease (PTLD) is a B cell proliferative disorder that is associated with Epstein-Barr virus (EBV), an ubiquitous herpesvirus. EBV-seronegative organ transplant recipients are at highest risk. EBV infection in PTLD lesions exists in a latent rather than lytic state, making tumor regression in response to antiviral agents unlikely. Viral latency proteins drive proliferation of T cells but also allow T cells to target PTLD lesions for destruction. Augmentation of the cellular immune response via the infusion of EBV specific cytotoxic T cells has yielded promising results in the prevention and treatment of PTLD in bone marrow transplant recipients. Efforts to extend this strategy to solid organ transplant recipients are ongoing. PMID- 11428991 TI - Cryptococcal meningitis in renal transplant patients associated with environmental exposure. AB - Fungal infections in renal transplant recipients are less common than bacterial infections; however, the morbidity from fungal infections is high. There is limited information in the literature concerning post-transplantation cryptococcal infection due to environmental exposure of patients living in high risk areas. We report three patients who were diagnosed with cryptococcal meningitis after kidney transplantation. Cryptococcal titers prior to transplant surgery were negative in all three patients. These patients all lived in rural areas and demonstrated evidence of environmental exposure leading to subsequent cryptococcal meningitis. All patients had exposure to pigeon and chicken excreta and, after treatment, two patients are alive and well with excellent allograft function. The third patient has marginal renal function but is currently not on dialysis. Early diagnosis is essential for salvage from these potentially lethal infections. Intense headache was a prominent feature in the clinical presentation of our patients, and should signal the need for early sampling and culture of spinal fluid. Meningismus was not present in any of our patients, even when other systemic symptoms were identified. We recommend a high index of suspicion post transplantation for all patients who may have environmental or occupational exposure to cryptococcus. If infection is detected quickly and treatment instituted promptly, patient recovery and allograft survival are possible. Long term therapy with fluconazole, a non-nephrotoxic agent, should permit eradication of the infection with preservation of kidney function. PMID- 11428992 TI - A 42-year-old lung transplant patient with ganciclovir-resistant cytomegalovirus (CMV) infection. PMID- 11428993 TI - Transplantation and the Second Law of Thermodynamics. PMID- 11428994 TI - Epidemiology of fungal infections in solid organ transplant patients. AB - The epidemiology of fungal infection in solid organ transplant patients is of concern due to the high mortality associated with this complication. Rates of fungal infections vary by type of transplant recipient. Most of these infections occur two to six months after transplantation. Liver transplant recipients are more likely to have early fungal infection which is often due to Candida species. Exogenous and endogenous Candida infection may occur in the immunosuppressed patient in the intensive care unit. Patients with chronic rejection are more likely to have late infection (after six months) which may be due to Aspergillus or endemic fungi such as Cryptococcus. Lung and heart-lung transplant recipients are more predisposed to infection with Aspergillus and other filamentous fungi, due to exposure of the transplanted organ to the external environment. Preventative measures such as environmental controls and chemoprophylaxis may be beneficial in high-risk patients. Emerging fungal pathogens such as the dematiaceous fungi may cause skin or soft tissue infection, or more serious systemic infections. Fungal infection should be ruled out in the solid organ transplant patient with early brain abscess. Characteristic risk factors in high risk types of solid organ transplant recipients should be recognized for early diagnosis and treatment of these infections associated with high morbidity and mortality. PMID- 11428995 TI - Fungal infections in patients undergoing blood and marrow transplantation. AB - Fungal infections are currently a leading cause of infectious morbidity and mortality in patients undergoing allogeneic blood and marrow transplantation (BMT). Although the introduction of azole antifungals for prophylaxis has had a significant impact on the incidence of candidal infections (especially those caused by C. albicans and C. tropicalis), invasive aspergillosis has increased in incidence in many centers worldwide. Given the long risk period corresponding with graft-versus-host disease, and the toxicities of currently available mold active antifungals, the development of a prevention strategy for these angioinvasive molds remains a challenge. The introduction of new antifungal drugs and adjunctive therapy to improve immune function may be beneficial in decreasing mortality associated with these infections in the future. Most importantly, a greater understanding of the pathogenesis of fungal disease and specific host risks is necessary to impact this increasingly important infection in immunocompromised hosts. PMID- 11428996 TI - Emerging fungal pathogens: evolving challenges to immunocompromised patients for the twenty-first century. AB - Opportunistic fungi have emerged during the past decade as important causes of morbidity and mortality in immunocompromised patients. Candida species constitute the third to fourth most common causes of nosocomial blood stream infections, and Aspergillus species have emerged as the most common infectious cause of pneumonic mortality in bone marrow/stem cell transplant recipients. Among HIV-infected patients, meningoencephalitis due to Cryptococcus neoformans ranks among the most common AIDS-defining infections. Hyaline septated filamentous fungi, such as Fusarium species, Acremonium species, Paecilomyces species, and Trichoderma species, are increasingly reported as causing invasive mycoses refractory to conventional therapy. Dematiaceous septated filamentous fungi, such as Pseudallescheria boydii, Bipolaris species, and Cladophialophora bantiana cause pneumonia, sinusitis, and CNS infection unresponsive to current therapy. An increasing number of different members of the class of Zygomycetes are reported as causing lethal infections, despite aggressive medical and surgical interventions. Yet the treatment for zygomycosis has not changed in approximately 40 years. The prevalence of the endemic mycoses, such as those due to Penicillium marneffei, Coccidioides immitis, and Histoplasma capsulatum, has been reported to expand rapidly in response to environmental exposures and increased numbers of vulnerable hosts in endemic regions of the world. Dermatophytoses are occurring with increasing prevalence and morbidity in elderly and immunocompromised patients. As we enter the next millennium, we may anticipate that emergent fungal infections will continue to develop in the settings of permissive environmental conditions, selective antifungal pressure, and an expanding population of immunocompromised hosts. PMID- 11428997 TI - Approaches to fungal diagnosis in transplantation. AB - The diagnosis of invasive fungal infection in patients undergoing solid organ or bone marrow transplantation remains a significant clinical challenge. Consideration of the epidemiology of these infections and host risk factors may be an important clue to a specific fungal diagnosis. Despite extensive investigation on methods such as serologic techniques to improve the rapid diagnosis of these infections, the diagnosis of invasive mycoses remains largely dependent on clinical presentation. For example, the signs and symptoms that result from angioinvasion of fungal organisms include pleuritic chest pain or hemoptysis. In a high-risk patient these findings can be important clues to invasive fungal infection. Cultures of opportunistic fungi in certain settings, such as Aspergillus in respiratory samples from immunosuppressed patients, may be associated with infection. Radiographic findings can also be useful to establish a diagnosis of infection. In patients with invasive aspergillosis as well as other angioinvasive moulds, chest CT scans may demonstrate lesions that are not visible on plain radiographs. Serodiagnosis of these infections remains largely investigational. Microbiological antifungal resistance has increasingly been reported, but in patients at high risk for serious fungal infection, including patients undergoing bone marrow and organ transplantation, antifungal resistance remains uncommon, particularly in Candida albicans. Higher doses of azoles should be used to treat patients with infections due to less susceptible yeasts and those with more serious infection. Prompt recognition of fungal infection combined with intensive antifungal therapy is needed for successful therapy. PMID- 11428998 TI - Amphotericin B and its new formulations: pharmacologic characteristics, clinical efficacy, and tolerability. AB - Amphotericin B (amB) remains the gold standard for the treatment of invasive fungal infections. However, the efficacy is limited, with response rates from 10% to 80%. Moreover, amB is toxic, especially for the kidneys. New formulations have been developed in an attempt to improve both efficacy and tolerability. In an attempt to reduce toxicity, a number of investigators have reconstituted amB in a lipid emulsion, but few data are available on efficacy in documented infections. An improvement in immediate and renal tolerance was obtained with equivalent daily dose regimens, but the therapeutic index does not appear to be improved. This approach cannot be recommended at present. Three lipid formulations have been developed and are now available in most countries: amB colloidal dispersion (ABCD), amB lipid complex (ABLC), and liposomal amB (AmBisome). The efficacy of ABCD on various fungal infections has been assessed in open trials, with a response rate of 49% in aspergillosis, 70% in candidiasis, and 67% in mucormycosis. In two randomized trials comparing ABCD with amB in invasive aspergillosis and in persistent febrile neutropenia, the response rates were equivalent. ABCD was less nephrotoxic. In contrast, immediate reactions to ABCD were as frequent and severe as with amB. These immediate effects are more frequent during the first infusions and lessen as treatment continues. The recommended dose is 3-4 mg/kg/day. ABLC appeared to be effective as rescue therapy in various types of invasive mycoses, with a response rate of 42% in aspergillosis, 67% in candidiasis, and 82% in fusariosis. Efficacy identical to that of amB was demonstrated in a comparative randomized trial involving patients with invasive candidiasis. General and renal tolerability is improved compared with amB. The recommended dose regimen is 5 mg/kg/day. Liposomal amB (AmBisome) is the only truly liposomal formulation. The response rates in preliminary trials were 66% in aspergillosis and 81% in candidiasis. Several comparative studies have confirmed that this formulation has similar or superior efficacy relative to amB in various fungal infections and also in the empirical treatment of febrile neutropenia. Renal and general tolerability is excellent. The optimal dosing remains unclear but is generally between 3 and 5 mg/kg/day. A double-blind trial comparing the tolerance of liposomal amB and ABLC demonstrated that both infusion related events and nephrotoxicity were significantly lower for liposomal amB. In sum, the new lipid formulations of amB are effective in various invasive fungal infections. The three formulations exhibit reduced nephrotoxicity compared with conventional amB. Large-scale comparative clinical trials may clarify issues of relative efficacy in various forms of mycotic infections. PMID- 11428999 TI - Listeria monocytogenes tricuspid valve endocarditis with septic pulmonary emboli in a liver transplant recipient. AB - Listeria monocytogenes has long been known as a pathogen of immunocompromised hosts, including solid organ and bone marrow transplant recipients. Its principal manifestations include bacteremia and meningitis. Endocarditis due to Listeria is far less common and in general affects the left side of the heart. We here report an unusual case of Listeria tricuspid valve endocarditis and septic pulmonary emboli in a sulfa-intolerant liver transplant recipient with a history of relapsing cytomegalovirus (CMV) hepatitis and an indwelling Hickman catheter. The literature on Listeria endocarditis and infections in transplant recipients is reviewed. The possible relationship between susceptibility to Listeria infection and the discontinuation of trimethoprim-sulfamethoxazole prophylaxis is of interest. PMID- 11429000 TI - Renal dysfunction and hyperglycemia in a renal transplant recipient. PMID- 11429001 TI - Prophylactic perioperative antibiotics. PMID- 11429002 TI - Pushing the envelope: transplantation and emerging infections. PMID- 11429003 TI - Prevention of recurrent cytomegalovirus disease in renal and liver transplant recipients: effect of oral ganciclovir. AB - BACKGROUND: Although the primary treatment of symptomatic cytomegalovirus (CMV) disease in organ transplant recipients is successful in >90% of individuals, relapsing disease, particularly in those with primary infection, remains an important problem. Previously, we had observed that the rate of symptomatic recurrence was >60% in those with primary disease (seronegative for CMV prior to transplant), and approximately 20% in those who were seropositive prior to transplant. The present study was undertaken to determine whether a maintenance regimen of oral ganciclovir for 2-3 months added to the routine 14-21 days of intravenous ganciclovir would further prevent symptomatic CMV recurrence. METHODS: From May 1995 until June 1998, all kidney and liver transplant recipients with confirmed tissue-invasive CMV disease or CMV syndrome were treated with 14-21 days of intravenous ganciclovir (5 mg/kg b.i.d. with dose adjusted for renal dysfunction) followed by 2-3 months of oral ganciclovir (2 g daily). The incidence of recurrence of CMV disease and/or viremia during and after oral therapy was then determined over a mean follow-up of 530.6 days. RESULTS: Thirty-seven patients, 19 kidney and 18 liver transplant recipients, were studied; 5 had biopsy-proven tissue-invasive disease (13.5) and 32 suffered a CMV syndrome (86.5). Twenty-one of these patients (58.6) were seronegative for CMV prior to transplant and received an allograft from a seropositive donor (D+/R ). Overall, 10 patients (27.0) developed CMV recurrence. Eight of 21 patients who were D+/R- for CMV (38.1) developed recurrence as opposed to 2 of 16 patients with other serologic status (12.5) (P=0.14). Patients with recurrent CMV disease and/or viremia had a peak antigenemia assay titer during their initial CMV event of 319.2 positive cells/2 slides compared with 109.8 positive cells/2 slides for patients without recurrent CMV infection (P=0.14); the trend of having a higher peak antigenemia assay titer among patients who recurred occurred both in patients who were at risk of primary CMV infection (D+/R- for CMV) and in those who were not. Two patients developed recurrent infection with strains of CMV that were resistant to ganciclovir. CONCLUSIONS: This new therapeutic regimen of oral ganciclovir following intravenous ganciclovir slightly reduced the overall rate of recurrent CMV disease and/or viremia, but it still did not adequately prevent CMV recurrence in patients who are at risk of primary infection prior to transplant. Of particular concern, 2 patients with primary infection treated with this regimen developed ganciclovir-resistant recurrent disease. PMID- 11429004 TI - Expression of porcine endogenous retrovirus in peripheral blood leukocytes from ten different breeds. AB - The expression of porcine endogenous retroviruses (PERV) was investigated in primary porcine peripheral blood leukocytes (PBL) of ten different pig breeds. The data suggest that PERV exists in all porcine PBL. A new retroviral element, a foamy-like pol-related sequence, was also detected in PBL. Three types of PERV were detected in almost every animal. The breeding of PERV-free pigs is likely to be difficult. Further studies are required to assess the infectious disease risks associated with xenotransplantation. PMID- 11429005 TI - Safety and efficacy of granulocyte colony-stimulating factor in kidney and liver transplant recipients. AB - BACKGROUND: Leukopenia is not infrequently encountered following solid organ transplantation, most often in the setting of cytomegalovirus (CMV) disease and/or its treatment with ganciclovir. The present study was undertaken to determine the safety and efficacy of granulocyte colony-stimulating factor (G CSF) in renal and liver transplant recipients with leukopenia. METHODS: Between 1 June 1991 and 1 June 1998, patients received G-CSF for 2 indications: 1) white blood cell count (WBC) < 3000/mm3, with a decline from baseline; 2) to shorten the duration of leukopenia associated with chemotherapy. A retrospective review of the outcome of such therapy was undertaken. RESULTS: 50 patients were given 100 courses of treatment with G-CSF; 35 of 168 liver transplant recipients (20.8%), 14 of 391 kidney transplant recipients (3.6%), and 1 of 4 recipients of combined liver-kidney transplants (25.0%) received from 1 to 9 courses of G-CSF. Presumed causes of leukopenia were identified as ganciclovir in 28 cases (28.0%), CMV in 21 (21.0%), chemotherapy in 12 (12.0%), sepsis in 11 (11.0%), azathioprine in 5 (5.0%), interferon in 3 (3.0%) and other causes in 20 cases (20.0%). The median length of therapy was 10.0 days (range 1-154 days) and the average dose of daily G-CSF received was 3.9+/-1.5 microg/kg/day. The average WBC was (2.4+/-1.3 )x 10(3)/microl at the beginning of therapy, and (13.8+/-9.1) x 10(3)/microl at the end of therapy. In 7 of 100 treatments (7.0%) a WBC of 5.0 x 10(3)/microl was not reached during G-CSF therapy; in 6 of these 7 cases, G-CSF therapy lasted fewer than 4 days. The mean time needed to reach a WBC count of 5 x 10(3)/microl was 3.7+/-3.3 days among 71 patients who had daily WBC counts sent. Eight G-CSF treatments (8.0%) were followed by episodes of rejection appearing during or within 2 months of treatment; 5 of them were biopsy-documented. No relation was found between the highest WBC obtained during G-CSF therapy and the risk of rejection. Eight patients (16.0%) died while receiving G-CSF, all from infection. Six of these 8 patients were receiving G-CSF for leukopenia secondary to sepsis. Overall, 25 patients (50.0%) received 49 courses of G-CSF secondary to CMV and/or ganciclovir therapy. In 40 of 49 courses (81.6%), ganciclovir could be continued at recommended doses. Twenty-one of 22 patients (95.5%) with symptomatic CMV infection had a clinical response to ganciclovir. Sixteen of 18 patients (88.9%) treated for a CMV infection and followed with serial antigenemia assays attained microbiological cure; both patients who did not were infected with ganciclovir resistant CMV. CONCLUSION: G-CSF was well tolerated in solid organ transplant recipients. It was particularly useful in patients with CMV disease, allowing optimal ganciclovir therapy. PMID- 11429006 TI - Investigation and control of aspergillosis and other filamentous fungal infections in solid organ transplant recipients. AB - Filamentous fungal infections are associated with high morbidity and mortality in solid organ transplant patients, and prevention is warranted whenever possible. An increase in invasive aspergillosis was detected among solid organ transplant recipients in our institution during 1991-92. Rates of Aspergillus infection (18.2%) and infection or colonization (42%) were particularly high among lung transplant recipients. Epidemiologic investigation revealed cases to be both nosocomial and community-acquired, and preventative efforts were directed at both sources. Environmental controls were implemented in the hospital, and itraconazole prophylaxis was given in the early period after lung transplantation. The rate of Aspergillus infection in solid organ transplant recipients decreased from 9.4% to 1.5%, and mortality associated with this disease decreased from 8.2% to 1.8%. The rate of Aspergillus infection or colonization among lung transplant recipients decreased from 42% to 22.5%; nosocomial Aspergillus infection decreased from 9% to 3.2%. Cases of aspergillosis in lung transplant recipients were more likely to be early infections in the pre-intervention period. Early mortality in lung transplant recipients decreased from 15% to 3.2%. Two cases of dematiaceous fungal infection were detected, and no further cases occurred after environmental controls. The use of environmental measures that resulted in a decrease in airborne fungal spores, as well as antifungal prophylaxis, was associated with a decrease in aspergillosis and associated mortality in these patients. Ongoing surveillance and continuing intervention is needed for prevention of infection in high-risk solid organ transplant patients. PMID- 11429007 TI - Visual symptoms after lung transplantation: a case of progressive multifocal leukoencephalopathy. AB - After solid organ transplantation, signs and symptoms of the central nervous system may present a diagnostic challenge. A 43-year-old patient developed a decrease in vision 15 months after bilateral lung transplantation. The initial diagnosis was a left posterior cataract, but left eye cataract extraction did not improve his vision. Seizures led to investigation of a broader differential diagnosis (cyclosporine intoxication, post-transplant lymphoproliferative disorder, infectious disease, chronic lymphatic leukemia). The clinical diagnosis of progressive multifocal leukoencephalopathy (PML) was confirmed by demonstration of JC virus in the cerebrospinal fluid and by autopsy findings. Modulation of the immunosuppressive regimen was unsuccessful. This case illustrates that decreased vision in immunocompromised patients may be the first manifestation of PML. PMID- 11429008 TI - Hepatic arterial thrombosis due to Mucor species in a child following orthotopic liver transplantation. AB - Infection is a frequent complication in patients following liver transplantation, and mycotic etiology is the third most common cause. Thrombosis of the hepatic artery is a high-risk condition for the graft and is generally secondary to noninfectious diseases. We present a 2-year-old child who developed hepatic artery obstruction due to Mucor sp. The child did not respond to medical treatment. We conclude that a high index of suspicion and rapid diagnosis are required so that aggressive and early treatment can be initiated. To our knowledge this is the first case published with this particular association. PMID- 11429009 TI - Fungal wound infection in a lung transplant recipient. PMID- 11429010 TI - Two further cases of tuberculosis in a renal allograft. PMID- 11429012 TI - Advances in the management of invasive fungal infections in organ transplant recipients: step by step. PMID- 11429013 TI - Amphotericin B preparations: a maximum tolerated dose in severe invasive fungal infections? AB - Availability of lipid formulations of amphotericin B has opened up the possibility of treating invasive fungal infections in immunocompromised patients with high doses of this antifungal agent. Evidence is emerging to suggest that lipid formulations may have heightened efficacy compared to conventional amphotericin B. The issue of optimal dosage has been a neglected area. This article reviews published data accrued from clinical, open-label, salvage, and other studies, and finds little support that the use of high doses of lipid formulations are more efficacious than lower doses. The response rates for invasive fungal infection from most studies are predictably around 56%, irrespective of the lipid formulation and dose used. Animal models provide evidence that low doses of a lipid formulation are as successful in reducing fungal dissemination and in prolonging survival as higher doses, although concomitant tissue fungal eradication is not as effectively achieved by the lower doses (survival-mycologic eradication dissociation). Kinetic studies performed in the clinically relevant setting of critically ill patients give further support to the use of low doses, since levels of liposomal amphotericin B at all dosages between 1 and 4 mg/kg/day are similar and above maximum inhibitory concentrations for commonly encountered fungi. There has only been one prospective randomised study designed with the primary end-point of comparing two dosages of an amphotericin B lipid formulation on clinical response and survival. That European Organization for Research and Treatment of Cancer (EORTC) study concluded that liposomal amphotericin B given at 1 mg was as efficacious as 4 mg/kg/day in treating neutropenic patients with invasive pulmonary aspergillosis. There are a multitude of unanswered questions concerning dosing, and their answers are confounded by difficulties in performing clinical trials and the multiplicity of factors other than antifungal chemotherapy that influence outcome. Maximum tolerated dose studies using existing lipid formulations, or perhaps with the newer formulations such as pegylated immunoliposomal amphotericin B, could be performed to shed light on this difficult area. PMID- 11429014 TI - Fluconazole in transplant recipients: options and limitations. AB - Fluconazole is currently a first-line agent used for therapy of non-critically ill patients with candidal infection. Its efficacy, the availability of an oral formula, and its relatively low toxicity make it a very attractive drug for use in many clinical situations. The advisability of prophylaxis and empirical treatment in transplant patients is a difficult issue for the following reasons: the potential emergence of resistance to the azoles, the lack of solid data establishing its advantage over placebo and/or oral nonabsorbable antifungal agents in some of the clinical conditions encountered, its ineffectiveness against molds, and its cost. Judicious use of fluconazole where its efficacy has been well established would provide the best therapy for patients and would limit the emergence of potential pathogens. As new antifungal agents are approved for clinical use, appropriate clinical trials will need to be designed and conducted in order for clinicians to make rational decisions in selecting the most appropriate drug for the specific indication. Prophylaxis and treatment with fluconazole in various transplant situations is reviewed. PMID- 11429015 TI - The use of itraconazole as prophylaxis against invasive fungal infection in blood and marrow transplant recipients. AB - Invasive fungal infections play a key role in contributing to morbidity and mortality in patients undergoing treatment for haematological malignancies and related diseases. Risk factors for development of invasive fungal infections after blood or bone marrow transplantation include the use of broad-spectrum antibiotics, steroids, mismatched or unrelated donor transplant, right atrial catheters, and prolonged or profound neutropenia. Previous attempts at use of oral itraconazole as antifungal prophylaxis in the setting of chemotherapy induced neutropenia were unsuccessful because of its poor absorption in capsule form. Itraconazole-cyclodextrin is well absorbed even in the presence of chemotherapy-induced neutropenia. Plasma levels of 250-500 ng/ml are required for prophylaxis. Studies to date show a favourable outcome in patients receiving itraconazole as prophylaxis against invasive fungal infections, although many studies looked at small numbers of patients and the incidence of invasive fungal infection in the control groups was low, prohibiting meaningful statistical evaluation. Fungi differ in their sensitivity to antifungal agents, and itraconazole is not the agent of choice in all patients. With the widespread use of antifungal prophylaxis, the possibility of resistance to antifungal agents and an increase in the number of invasive fungal infections caused by ubiquitous fungi previously considered nonpathogenic must be considered as potential problems. PMID- 11429016 TI - Microbiological diagnosis of invasive fungal infections in transplant recipients. AB - Invasive fungal infections remain an important cause of morbidity and mortality in transplant recipients. Since conventional diagnostic tools such as culture lack sensitivity and specificity, alternative diagnostic assays have been developed. Among the most promising techniques are the detection of fungal DNA and serology. Fungal DNA can be detected with high sensitivity and specificity when performed with specimens from sterile sites such as blood. Polymerase chain reaction (PCR) assays can be used to detect a broad range of fungal pathogens and combined with species identification. Multicenter diagnostic studies are needed to establish the diagnostic value of PCR but lack of standardization hampers such studies. The serodiagnosis of invasive fungal infections has become an important tool in the management of invasive fungal infections. Especially the detection of circulating galactomannan has been shown to be a sensitive marker for invasive aspergillosis. Both serology and PCR can be used to monitor the response to antifungal therapy. The optimal use of non-culture-based methods is in prospective screening of patients at high risk. Since the incidence of disease greatly influences the positive predictive value, screening should take place only in those patients at very high risk for invasive fungal disease. PMID- 11429017 TI - A strategy for managing fungal infections in haematopoietic stem cell transplantation. AB - As more indications continue to be found for allogeneic haematopoietic transplantation, more patients are at risk for invasive fungal infectious diseases (IFID), particularly candidiasis and aspergillosis. Risk factors for disease are becoming better defined and diagnostic methods have improved considerably. In addition, there is now international agreement that three elements form the basis for defining IFID (host factors, clinical evidence, and mycological results), that imaging is acceptable for diagnosing disease, and that indirect tests such as antigen detection are also adequate mycological proof of cause. There are also more drugs available and still more to come, offering the potential for selective prophylaxis as well as preemptive and specific therapy, as well as for flexible administration. Hence, all the elements are in place for designing and testing an effective and economically sound strategy for dealing with IFID. PMID- 11429018 TI - Atypical presentation of Madurella mycetomatis mycetoma in a renal transplant patient. AB - A renal transplant patient presented with a swelling on his foot. Fungal hyphae were found in the abscess, and culture demonstrated Madurella mycetomatis, an agent of mycetoma. There were no black grains in the pus and no overlying sinuses. This was an atypical presentation of maduromycosis in a renal transplant patient. PMID- 11429019 TI - Prevention of cytomegalovirus infection in organ transplant recipients. PMID- 11429020 TI - Infections of the central nervous system in transplant recipients. AB - Central nervous system (CNS) infections, accounting for 4-29% of CNS lesions in transplant recipients, are a significant post-transplant complication. Focal CNS infectious lesions or brain abscesses have been documented in 0.36-1% of the transplant recipients. Mycelial fungi, particularly Aspergillus, are by far the most frequent etiologies of post-transplant brain abscesses. Bacteria, with the exception of Nocardia, are rarely associated with brain abscesses in transplant recipients. Time of onset and concurrent extraneural lesions have implications relevant towards invasive diagnostic procedures in transplant recipients with brain abscesses. Meningoencephalitis in transplant recipients is predominantly due to viruses, e.g., herpesviruses, and less frequently due to Listeria monocytogenes, Toxoplasma gondii, and Cryptococcus. Despite a wide, and at times perplexing array of opportunistic pathogens that can cause CNS infections, the temporal association of the infection with the time elapsed since transplantation, risk factors, clinical manifestations, and neuroimaging characteristics of the lesion can allow a reasoned and rational approach towards the recognition, diagnosis, and appropriate management of CNS infections in transplant recipients. PMID- 11429021 TI - Prevention of primary cytomegalovirus disease in organ transplant recipients with oral ganciclovir or oral acyclovir prophylaxis. AB - BACKGROUND: Optimal prophylaxis against cytomegalovirus (CMV) disease for organ transplant patients at risk for primary infection (donor seropositive, recipient seronegative, D+R-) remains to be determined. We hypothesized that prolonged oral ganciclovir therapy following intravenous therapy would provide increased protection. METHODS: A total of 155 evaluable D+R- organ transplant recipients from 13 transplant centers were entered into the study: all received intravenous ganciclovir (5 mg/kg/day) for 5-10 days and then either oral acyclovir (400 mg tid) or oral ganciclovir (1 g tid) for an additional 12 weeks. Patients were assigned to their treatment groups at a central randomization site, with a separate randomization scheme for each of the organs transplanted (kidney, heart, or liver). In the case of kidney transplants, the patients were stratified according to source of the kidney (living related vs. cadaveric donor). The primary endpoint was the incidence of CMV disease in the first six months post transplant. RESULTS: Treatment with oral ganciclovir was associated with a significant decrease in the incidence of symptomatic disease or viremia when compared with the oral acyclovir group (32% vs. 50%, P<0.05). This difference was most marked in terms of tissue invasive disease: only 3 of 15 symptomatic patients in the ganciclovir group vs. 10 of 21 in the acyclovir group developed tissue-invasive infection (P<0.05). There was a significant difference in the time to CMV disease or viremia in the two groups: mean time 212+/-17 days post transplant for the acyclovir group vs. 291+/-13 days for the ganciclovir group (P<0.001). The incidence of allograft rejection was 34% in the ganciclovir group and 46% in the acyclovir group (P=NS). Leukopenia was more common in the ganciclovir group (P<0.05), but in no case did it require drug discontinuation. Ganciclovir resistance did not develop in this study. CONCLUSION: Prophylaxis with oral ganciclovir following a brief course of intravenous ganciclovir provides useful protection against primary CMV disease. PMID- 11429022 TI - Preemptive treatment approach to cytomegalovirus (CMV) infection in solid organ transplant patients: relationship between compliance with the guidelines and prevention of CMV morbidity. AB - Cytomegalovirus (CMV) remains a major cause of morbidity in solid organ transplant patients. In order to reduce CMV morbidity, we designed a program of routine virological monitoring that included throat and urine CMV shell vial culture, along with peripheral blood leukocyte (PBL) shell vial quantitative culture for 12 weeks post-transplantation, as well as 8 weeks after treatment for acute rejection. The program also included preemptive ganciclovir treatment for those patients with the highest risk of developing CMV disease, i.e., with either high-level viremia (>10 infectious units [IU]/106 PBL) or low-level viremia (<10 IU/106 PBL) and either D+/R- CMV serostatus or treatment for graft rejection. During 1995-96, 90 solid organ transplant recipients (39 kidneys, 28 livers, and 23 hearts) were followed up. A total of 60 CMV infection episodes occurred in 45 patients. Seventeen episodes were symptomatic. Of 26 episodes managed according to the program, only 4 presented with CMV disease and none died. No patient treated preemptively for asymptomatic infection developed disease. In contrast, among 21 episodes managed in non-compliance with the program (i.e., the monitoring was not performed or preemptive treatment was not initiated despite a high risk of developing CMV disease), 12 episodes turned into symptomatic infection (P=0.0048 compared to patients treated preemptively), and 2 deaths possibly related to CMV were recorded. This difference could not be explained by an increased proportion of D+/R- patients or an increased incidence of rejection among patients with episodes treated in non-compliance with the program. Our data identify compliance with guidelines as an important factor in effectively reducing CMV morbidity through preemptive treatment, and suggest that the complexity of the preemptive approach may represent an important obstacle to the successful prevention of CMV morbidity by this approach in the regular healthcare setting. PMID- 11429023 TI - Prospective study of toxoplasma reactivation by polymerase chain reaction in allogeneic stem-cell transplant recipients. AB - Toxoplasmosis is a rare but life-threatening complication of allogeneic stem-cell transplantation. Polymerase chain reaction (PCR) offers the possibility to make the diagnosis earlier than conventional techniques, and is then expected to improve the prognosis. We undertook a prospective screening using a competitive PCR in blood in 32 stem-cell transplant recipients. The sampling covered the first 150 days post-transplant, at days 21, 30, 45, 60, 90, 120, and 150. Twenty four patients had anti-toxoplasma antibodies before transplant. Three of them (12.5%) had transient PCR-positive samples at 21, 45, and 90 days post transplant, respectively. The three PCR-positive patients were febrile but had no funduscopic examination or cerebral computerised tomography (CT) scan abnormalities. The PCR signal disappeared when the patients were given trimethoprim-sulfamethoxazole, and no full-blown toxoplasmosis was observed. Toxoplasma reactivation evidenced using PCR is frequent in seropositive patients not receiving trimethoprim-sulfamethoxazole during the 1-3 months post transplant. Toxoplasma PCR should be included in the diagnostic strategy of fever of unexplained origin in allogeneic stem-cell transplant recipients. Then, prompt specific therapy can be initiated to avoid development of full-blown toxoplasmosis. PMID- 11429024 TI - Significant post-transplant hypogammaglobulinemia in six heart transplant recipients: an emerging clinical phenomenon? AB - BACKGROUND: The recent development of powerful agents such as mycophenolate mofetil and tacrolimus has altered current regimens for the prevention and treatment of allograft rejection. Questions have been raised about these newer regimens in terms of susceptibility to opportunistic infections and effects on host defenses. Severe hypogammaglobulinemia has been infrequently described in solid organ transplant recipients, but has been recently noted in six heart transplant recipients at one center, of whom five were receiving a combination of tacrolimus, mycophenolate mofetil, and prednisone. METHODS: Case summaries of six recent heart transplant recipients with total immunoglobulin G (IgG) levels of less than 310 mg/dl, five of whom had cytomegalovirus (CMV) infection and three of whom had multiple infections including Nocardia, invasive Trichophyton, and Acinetobacter bacteremia. Previous literature was reviewed with the aid of a Medline search using the search terms hypogammaglobulinemia; kidney, liver, heart, lung, and organ transplantation; mycophenolate mofetil; tacrolimus; cyclosporine; azathioprine; and nocardiosis. RESULTS: We here report six cardiac transplant recipients seen over a period of one year who were found to have immunoglobulin G levels of 310 mg/dl or below (normal: 717-1400 mg/dl). The first five patients were diagnosed because of evaluation for infections; the sixth, who was asymptomatic with an IgG level of 175, was found during screening for hypogammaglobulinemia instituted as a result of these first five patients. All six patients had received steroid pulses for rejection; all received mycophenolate mofetil; and 5/6 had been switched from cyclosporine to tacrolimus because of steroid-resistant rejection. Transient neutropenia (absolute neutrophil count less than 1000) was observed in 2/6; 3/6 had received OKT3 therapy for refractory rejection. These six patients were treated with a combination of antimicrobials, immunoglobulin replacement, and decrease in immunosuppressive therapy. CONCLUSION: The finding of unexpected hypogammaglobulinemia and concomitant infectious complications in six heart transplant recipients highlights a possible complication in a subset of patients receiving newer immunosuppressive agents. A larger prospective study is underway to determine risk factors for development of post-transplant hypogammaglobulinemia and to assess pre-transplant immune status of these recipients. Monitoring of immunoglobulin levels in high-risk patients receiving intensified immunosuppressive therapy for rejection may help to prevent infectious complications. PMID- 11429025 TI - Antigen detection in the diagnosis and management of a patient with probable cerebral aspergillosis treated with voriconazole. AB - This report describes the diagnosis and management of a 16-year-old boy who developed neurological signs and symptoms suggestive of cerebral aspergillosis following a haploidentical bone marrow transplant. A new sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of Aspergillus galactomannan circulating antigens (Platelia Aspergillus, Sanofi Diagnostic Pasteur, France) was used on serum and cerebrospinal fluid to obtain a presumptive diagnosis and to monitor the course of the disease. Having failed conventional therapy with amphotericin B, the patient received compassionate treatment with voriconazole for a period of 37 days. High levels of voriconazole were observed in both serum and cerebrospinal fluid (CSF), with a trend toward accumulation. After 7 days a marked improvement in the patient's neurological symptoms was noted, and ELISA data indicated a corresponding decrease in Aspergillus galactomannan levels in both serum and CSF. Voriconazole was well tolerated, with only transient increases in ALT/AST recorded during therapy. Although the patient survived the acute Aspergillus infection, he subsequently died of an unrelated infection. PMID- 11429026 TI - Cerebral toxoplasmosis and Guillain-Barre syndrome after allogeneic peripheral stem cell transplantation. AB - We report an unusual case of cerebral toxoplasmosis associated with Guillain Barre syndrome (GBS) in a 25-year-old patient diagnosed with chronic myelogenous leukaemia (CML), who underwent a mismatched allogeneic peripheral stem cell transplantation (PSCT). On day +83 he started with fever, and 7 days later tremor, muscular weakness, diplopia, dysarthria, respiratory difficulty, and universal arreflexia appeared, compatible with GBS. As the patient had a positive cytomegalovirus (CMV) antigenemia, this was the aetiology suspected for his neurologic findings, but specific treatment failed to improve his clinical situation, and he died on day +123. Necropsy demonstrated cerebral toxoplasmosis and axonal degeneration of nerve roots compatible with the axonal form of GBS. Interestingly, the polymerase chain reaction (PCR) signal for Toxoplasma gondii in two different cerebrospinal fluid (CSF) samples had been negative. In addition, this case showed unique magnetic resonance imaging (MRI) abnormalities. We conclude that a negative PCR on CSF cannot exclude toxoplasmosis in a transplant patient, and we emphasise the importance of considering Toxoplasma as an aetiology of fever and neurological symptoms in the transplant setting. PMID- 11429027 TI - Control of hepatitis in the transplant patient: a journey begun--but not there yet. PMID- 11429028 TI - Hepatitis B in transplantation. AB - Hepatitis B has been a major challenge within the field of transplantation over the past few decades. Due to aggressive recurrence post-transplant, patients with hepatitis B have been excluded from the benefits of both solid organ and bone marrow transplants. Progress has been made, however, through an improved understanding of the biology of hepatitis B and the development of new antiviral strategies that can reliably suppress the virus. Patients with hepatitis B are now candidates for transplantation in an increasing number of circumstances. Careful pre-transplant evaluation is mandatory, together with a tailored antiviral regimen depending on the replicative status of the virus and the organ being transplanted. Minimizing steroid dose following transplantation is an important part of the strategy to reduce the risk of viral reactivation. Lamivudine has been an important development and it has assumed an increasing role in the management of these patients. As additional antivirals are developed, increasingly effective drug combinations will prevent viral recurrence as well as the emergence of drug-resistant mutants, which plagues the use of single agents. It is a rapidly evolving field and there is every reason for continued optimism. PMID- 11429029 TI - Effects of immunosuppression and organ transplantation on the natural history and immunopathogenesis of hepatitis C virus infection. AB - The hepatitis C virus (HCV) is recognized as the leading cause for parenterally transmitted hepatitis. It is characterized by a high propensity to chronicity. Several efforts have been directed towards understanding the natural history of chronic HCV infection and the immunopathogenic pathways involved in mediating liver injury in the non-immunosuppressed and immunosuppressed states. In the non immunosuppressed setting, liver damage seems to be largely immune mediated. In contrast, in the non-immunosuppressed state, there are several other factors that may modify the natural course of the infection and play a role in mediating liver injury. In this review we will address the natural history, virological and immunological aspects of HCV infection. Also, the role played by immunosuppression and organ transplantation in modifying the course of the infection and the pathogenesis of liver injury will be discussed. PMID- 11429030 TI - Allograft dysfunction and hyperbilirubinemia in a liver transplant recipient. PMID- 11429031 TI - Invasive Scedosporium apiospermum infection in a heart transplant recipient presenting with multiple skin nodules and a pulmonary consolidation. PMID- 11429032 TI - The use of antimicrobial drugs in transplant patients. PMID- 11429033 TI - Diagnostic yield of bronchoalveolar lavage following renal transplantation. AB - Organ transplant recipients are at high risk of infectious pulmonary complications. In this retrospective study, the diagnostic yield of bronchoalveolar lavage (BAL) was evaluated in renal transplant recipients. The results were analysed in special regard to the clinical presentation of pulmonary infections and the possible impact of new immunosuppressive agents. Over a 5-year period 91 BAL were performed in 71 renal transplant recipients. Microorganisms were isolated from 69% of BAL (63/91): bacteria 32%; cytomegalovirus (CMV) 27%; Pneumocystis carinii (PC) 22%; other viruses 9% (HSV; EBV, RSV, adenovirus, HHV8); Aspergillus fumigatus 1%. Total cell counts and neutrophil counts in BAL were significantly elevated in bacterial infection, whereas BAL positive for PC showed eosinophilia (P<0.05). There was no association between clinical symptoms and the radiological pattern of infiltrates and the type of infection. Immunosuppression containing tacrolimus or mycophenolate mofetil was associated with a significantly higher percentage of PC and CMV infections compared to cyclosporin-based immunosuppression (65% vs. 30%, P<0.005). A considerable number of PC and CMV infections occurred beyond 6 months after transplantation. In conclusion, BAL has a high diagnostic yield in renal transplant recipients. Infection with CMV and PC should also be considered beyond 6 months after transplantation, and prophylaxis for opportunistic infections should be given if the immunosuppression is intensified. PMID- 11429034 TI - Patterns of cytomegalovirus infection in simultaneous kidney-pancreas transplant recipients receiving tacrolimus, mycophenolate mofetil, and prednisone with ganciclovir prophylaxis. AB - BACKGROUND: The impact of tacrolimus (TAC), mycophenolate mofetil (MMF) and steroid immunosuppression on cytomegalovirus (CMV) infection in combination with ganciclovir prophylaxis in simultaneous kidney-pancreas transplantation (SKPT) has not been well studied. METHODS: A retrospective analysis was made of 75 SKPTs performed between 1 January 1996 and 7 January 1999. All patients received ganciclovir for 3 months, but CMV donor (D)+ / recipient (R)- patients received ganciclovir for 6 months. RESULTS: 16/74 (22%) were CMV D+/R-, 25 (33%) D+/R+, 16 (22%) D-/R+, and 17 (23%) D-/R- (1 patient with unknown donor serology was excluded). The mean time to CMV infection was 198 days post-transplant. The incidence of either CMV infection or tissue invasive CMV disease was 16/74 (22%), including 9 (12%) with CMV infection and 7 (10%) CMV disease. The one-year patient, kidney, and pancreas graft survival rates were 91%, 89%, and 83%, respectively. The mean follow-up was 29 months (minimum of 12 months). CMV infection was not associated with an increased incidence of graft failure or mortality. The D+/R- group had the highest incidence of CMV infection (44%) compared with the other serologic groups (17%, P=0.02). Concurrent CMV and rejection occurred more frequently in the D+/R- than the other serologic groups (25% vs. 7%, P=0.03). The D-/R- group had the best outcomes, with no CMV infection, improved kidney graft survival at the end of follow-up (82% vs. 72%, P=0.04) and the highest event-free survival (no CMV infection, rejection, or graft loss) when compared to the other groups (76% vs. 33%, P<0.01). CONCLUSIONS: Compared to previous studies, ganciclovir prophylaxis delayed the onset and reduced the severity of CMV infection in patients receiving TAC, MMF, and steroids. Despite ganciclovir prophylaxis, CMV seronegative patients receiving CMV D+ organs had worse outcomes than seronegative recipients receiving CMV D- organs. PMID- 11429035 TI - Famciclovir treatment of hepatitis B infection following liver transplantation: a long-term, multi-centre study. AB - Famciclovir is a novel guanosine nucleoside analogue with activity against herpes viruses and hepatitis B virus (HBV). Several preliminary reports have described efficacy of famciclovir in patients with recurrent hepatitis B after orthotopic liver transplantation (OLT). This report describes the largest study to date of long-term famciclovir treatment in patients with de novo or recurrent hepatitis B post-OLT. One hundred thirty patients with detectable serum HBV DNA after OLT received oral famciclovir 500 mg tid on a compassionate-use basis. Safety analyses included all treated patients; efficacy was assessed in all patients and a subgroup of 73 patients with complete baseline HBV DNA and alanine aminotransferase (ALT) data who had received > or =6 months of treatment. Efficacy parameters included serum levels of HBV DNA, ALT, and anti-HBe or anti HBs seroconversion rates. Of the 70 patients treated for > or =6 months who could be evaluated for response/non-response to famciclovir, 52 (74%) were responders, defined as patients who experienced a 70% decrease or more in HBV DNA levels from baseline, or who became HBV DNA-negative, for at least two consecutive visits. In famciclovir responders, HBV DNA levels decreased by a median of 91% after 12 weeks of treatment, 95% after 6 months and >99% after 18 months of treatment. Marked differentiation between responders and non-responders could be made soon after the onset of treatment. Among anti-HBe positive patients with evidence of HBV replication, 12/13 were responders. Patients with high baseline ALT levels experienced more rapid suppression of HBV DNA during therapy with famciclovir. Famciclovir therapy was safe and well tolerated; serious adverse events were reported infrequently. Famciclovir treatment may be beneficial in patients with hepatitis B infection post-OLT. PMID- 11429036 TI - Disseminated toxoplasmosis after bone marrow transplantation: report of 9 cases. AB - Toxoplasma infection following bone marrow transplantation (BMT) is infrequently reported. We report 9 cases of disseminated Toxoplasma gondii infection in BMT recipients documented during an 11-year period at our institution. The incidence of T. gondii infection in our institution (1.14 per 100 allogeneic BMT) is higher than previously reported. The most frequently affected sites were the brain, lungs, and heart. Findings common to most patients who developed toxoplasmosis were positive pre-transplant serology, allogeneic transplant and graft-versus host disease and its treatment, as well as BMT from matched unrelated donors. All 9 patients died and 8 were diagnosed only after autopsy. Heightened awareness of the occurrence of toxoplasmosis in marrow recipients, especially in highly endemic areas, and early diagnosis and therapy are needed for a better outcome. PMID- 11429037 TI - Fatal Salmonella group G enteritis mimicking intestinal graft-versus-host disease in a bone marrow transplant recipient. PMID- 11429038 TI - Human granulocytic ehrlichiosis in pancreas transplant recipients. AB - Human ehrlichioses are tick-borne infections caused by bacteria in the genus Ehrlichia of the family Rickettsiaceae. To date there have been three cases of ehrlichiosis reported in the transplant population, a human monocytic ehrlichiosis (HME) infection in a liver transplant recipient and two cases of human granulocytic ehrlichiosis (HGE) in kidney transplant recipients. We report three pancreas transplant patients who developed HGE in the last two years at a single southeastern center in the United States. All three patients had clinical, laboratory, and pathophysiologic findings on bone marrow biopsy and peripheral blood smears consistent with HGE, and responded to doxycycline therapy. In the setting of potent immunosuppression, ehrlichiosis should be considered in the differential diagnosis of transplant patients presenting with persistent fever, pancytopenia, and abnormal liver function. Patients with ehrlichiosis infection may be at risk for developing other opportunistic infections or lymphoproliferative disease. PMID- 11429039 TI - Disseminated Dactylaria constricta infection in a renal transplant recipient. AB - We report the case of a 32-year-old renal transplant recipient who developed disseminated Dactylaria constricta infection. The patient died despite treatment with amphotericin B, itraconazole, and fluconazole. PMID- 11429040 TI - Cidofovir treatment of human polyomavirus-associated acute haemorrhagic cystitis. AB - We report the case of an 18-year-old patient who received an allogeneic bone marrow transplant from an HLA-identical unrelated donor for a Ph+ acute lymphoblastic leukemia, in his third complete remission. Cyclophosphamide and busulfan were used as conditioning treatment. Acute graft-versus-host disease developed on day +9, and the response to adequate treatment (steroids) was favourable. On day +45 the patient developed an acute severe haemorhragic cystitis, and BK polyomavirus was demonstrated in urine samples using electron microscopy and polymerase chain reaction. Urinary symptoms did not improve in spite of palliative treatment, but a response was evident after 2 weeks of cidofovir treatment. PMID- 11429041 TI - Cytomegalovirus maculopapular eruption in a kidney transplant patient. AB - Cytomegalovirus (CMV) is the most important viral agent in kidney transplantation. Clinical manifestations of CMV disease in transplantation include hepatitis, pneumonitis, pancreatitis, kidney allograft dysfunction, colitis, and meningoencephalitis. However, skin involvement is rare. We describe a severely compromised cadaveric-kidney transplant recipient who developed renal failure, colonic ulcers, and a maculopapular rash accompanied by fever and malaise 4 months after transplantation. Only the skin biopsy was diagnostic and consistent with CMV disease. Intravenous ganciclovir administration resulted in clinical improvement of CMV-induced skin lesions; kidney function normalized and the patient became asymptomatic after 14 days of ganciclovir therapy. Nephrologists should consider the diagnosis of CMV disease in the febrile immunosuppressed patient with skin involvement. Skin biopsy must be considered as a useful and safe procedure in patients with a rash to obtain a prompt diagnosis and efficiently treat this immunocompromised population. PMID- 11429042 TI - Management of relapsing cytomegalovirus disease in transplant recipients: diagnostic and therapeutic dilemmas. PMID- 11429043 TI - pncA mutations in clinical Mycobacterium tuberculosis isolates from Korea. AB - BACKGROUND: Pyrazinamide (PZA) is among the first-line drugs for the treatment of tuberculosis. In vitro, it kills semidormant mycobacteria only at low pH. The purpose of this study was to compare PZA resistance with pyrazinamidase (PZase) activity and the genotype to better understand the molecular basis of PZA resistance and to expand the profile of pncA mutations worldwide. RESULTS: Of the 28 tested strains of Mycobacterium tuberculosis, 6 were susceptible to PZA and positive for PZase activity and had no pncA mutations. Twenty-one strains were resistant to PZA and negative for PZase activity and had mutations in the pncA gene, including 15 point mutations, 5 insertions, and 2 deletions. One strain had no mutation in the pncA gene, even though it was resistant to PZA and negative for PZase activity. Three isolates had adenine to guanine point mutations in the 11 upstream region, making this the most common type of pncA mutations in this study, with at least two different RFLP patterns. CONCLUSION: These data help in the understanding of the molecular basis of PZA resistance. An adenine to guanine point mutation in the -11 upstream region was the most common type of pncA mutation in our isolates. The results of pncA mutation analyses should be carefully interpreted for epidemiologic purposes. PMID- 11429044 TI - Frameshift mutations at mononucleotide repeats in RAD50 recombinational DNA repair gene in colorectal cancers with microsatellite instability. AB - To identify additional genes targeted for microsatellite instability (MSI), we search for human genes which contain mononucleotide repeats in their coding region, selected 7 genes (RAD50, DNA-PKcs, FLASH, Apaf-1, XPG, CtIP, and MLSN1), and analyzed frameshift mutations in them. Here we report that 60% (3 out of 5) of human colorectal cancer cell lines exhibiting a high frequency of MSI (MSI-H) and 46% (6 out of 13) of MSI-H primary colorectal tumors had mutations in the (A)9 repeat of RAD50 recombinational repair gene. In contrast, no frameshift mutations were found in any of the 5 MSI-negative colorectal cancer cell lines, 8 colorectal tumors exhibiting a low frequency of MSI (MSI-L), or 28 MSI-negative colorectal tumors. No mutations were found in the mononucleotide repeats of 6 other genes, even in MSI-H cancers. These results suggest that RAD50 frameshift mutations may play a role in the tumorigenesis of MSI-H colorectal cancers. PMID- 11429045 TI - Association of ANA, a member of the antiproliferative Tob family proteins, with a Caf1 component of the CCR4 transcriptional regulatory complex. AB - A 35-kDa protein, ANA, belongs to an emerging family of antiproliferative proteins consisting of Tob, Tob2, ANA / BTG3, PC3B, PC3 / TIS21 / BTG2, and BTG1. All of these, except ANA and PC3B, have been shown to interact with the CCR4 transcription factor-associated protein Caf1. Here we show that ANA also associates with Caf1, ANA being the preferred partner of Caf1 among the Tob family proteins. Although ANA is likely to interact with Caf1 at its amino terminal half, which is conserved among the family members, our data suggest that the carboxyl-terminal half of ANA plays a role in the interaction. Finally, in situ hybridization experiments revealed that expression of Caf1 overlaps at least in part with that of ANA. Thus, ANA could function through its interaction with Caf1. PMID- 11429046 TI - Reversal of cisplatin resistance by the 1,4-benzothiazepine derivative, JTV-519. AB - The 1,4-benzothiazepine derivative JTV-519 is a new type of calcium ion channel modulator. We examined the modulatory effect of JTV-519 on the antitumor activity of several platinum compounds (cisplatin, carboplatin, and nedaplatin) in a human cancer cell line resistant to cisplatin (PC-14 / CDDP) in vitro. PC-14 / CDDP cells showed 8-fold resistance to cisplatin compared with the parental PC-14 cells as determined by dye formation [3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide, MTT] assay. In PC-14 / CDDP, but not PC-14 cells, augmentation of cytotoxicity was observed when a nontoxic concentration (10 mM) of JTV-519 was combined with the platinum compounds. Increased intracellular cisplatin accumulation was observed in PC-14 / CDDP cells in the presence of JTV 519 as measured by atomic absorption assay. Therefore, increased cisplatin accumulation was considered to be a possible mechanism underlying the reversing effect of JTV-519 on cisplatin resistance. These results suggest that JTV-519 is a potent agent reversing cisplatin resistance. PMID- 11429047 TI - Chemopreventive effects of scordinin on diethylnitrosamine and phenobarbital induced hepatocarcinogenesis in male F344 rats. AB - Modifying effects of scordinin, a biological active component in garlic, on diethylnitrosamine (DEN)- and phenobarbital (PB)-induced hepatocarcinogenesis were examined in rats. Male F344 rats, 5 weeks old, were divided into 8 groups. After a week, groups 1 - 5 were given DEN (100 mg / kg body weight, i.p.) once a week for 3 weeks, whereas groups 6 - 8 received vehicle treatment. Group 2 was given 600 ppm scordinin-containing diet in the initiation phase. From 4 weeks after the start of experiment, groups 3 and 5 were fed scordinin, and groups 1 - 3 and 7 received drinking water containing 500 ppm PB. Group 6 was given scordinin diet alone throughout the experiment (24 weeks). The incidences of hepatocellular adenoma and carcinoma were significantly smaller in group 3 than those in group 1 (P < 0.005 and P < 0.05, respectively). The average numbers of liver neoplasms in groups 2 and 3 were significantly smaller than in group 1 (P < 0.01 and P < 0.0001, respectively). Glutathione S-transferase placental form positive foci were also significantly decreased by scordinin treatment in the initiation or promotion phase. Scordinin significantly decreased the mean number of nucleolar organizer regions' protein (AgNORs) / nucleus in hepatocellular adenoma and carcinoma. AgNORs / nucleus in the non-lesional area was also significantly decreased by scordinin treatment during the promotion phase. These results suggest that scordinin is a promising chemopreventive agent for liver neoplasia. PMID- 11429048 TI - Five quantitative trait loci affecting 4-nitroquinoline 1-oxide-induced tongue cancer in the rat. AB - In our previous study, Dark-Agouti (DA) rats were found to be highly susceptible to 4-nitroquinoline 1-oxide (4NQO)-induced tongue carcinoma (TC), whereas Wistar / Furth (WF) rats were barely susceptible. Interval mapping analysis of reciprocal backcross rats showed two quantitative trait loci (QTL) on rat chromosomes (RNO) 1 and 19. In the present study, a composite interval mapping analysis was applied to 4NQO-induced TC in 130 (DA x WF) F2 rats, demonstrating five independent QTL, Tongue squamous cell carcinoma 1 - 5 (Tscc1 - 5), responsible for phenotypic differences in the size and number of TCs in the two strains. Two of these QTL were mapped on RNO1, and the others were mapped on RNO4, 14, and 19. The DA allele at these loci consistently yielded semidominant susceptibility to TC. Out of the five loci detected in this F2 generation, Tscc1 and 2 were identical to Stc1 and Rtc1 described in our previous study, but the other three were novel. We propose a new nomenclature consistent with their function. Genome-wide screening of the F2 progeny also suggested the presence of three additional QTL on RNO5, 6, and 10. The possible roles of these loci in tongue carcinogenesis are discussed. PMID- 11429049 TI - Suppression of occurrence and advancement of beta-catenin-accumulated crypts, possible premalignant lesions of colon cancer, by selective cyclooxygenase-2 inhibitor, celecoxib. AB - Suppression of occurrence and advancement of premalignant lesions is important for cancer prevention. Our previous studies clarified that beta-catenin accumulated crypts, independent of aberrant crypt foci (ACF), are probably direct precursors of colon cancers in rats. Here we investigated the effects of a selective cyclooxygenase-2 inhibitor, celecoxib, on the development of beta catenin-accumulated crypts in comparison with those on ACF. Male F344 rats were divided into 4 groups. Groups 1 - 3 were administered azoxymethane (AOM) s.c. at a dose of 15 mg / kg body weight, once weekly for 3 weeks to induce beta-catenin accumulated crypts. Groups 2 and 3 also received experimental diet containing celecoxib (500 and 1500 ppm, respectively) for 8 weeks, starting a week before the first dosing of AOM. At termination, the frequency and crypt multiplicity (number of crypts / lesion) of beta-catenin-accumulated crypts of groups 2 and 3 were significantly less than that of group 1. Furthermore, numbers of silver stained nucleolar organizer regions (AgNORs) / nucleus in beta-catenin accumulated crypts were also decreased by exposure to celecoxib. In this study, celecoxib had greater effects on the frequency and growth of beta-catenin accumulated crypts than on those of ACF. These findings represent additional evidence that beta-catenin-accumulated crypts are premalignant lesions of colon cancer. The results also suggest that beta-catenin-accumulated crypts could be a novel target for evaluation of possible chemopreventive agents against colon carcino-genesis, and indicate that possible chemopreventive effects of celecoxib on the initial stage of colon carcinogenesis may be related to modulation of cell proliferation activity in such early lesions. PMID- 11429050 TI - Quantitative analysis of BRCA1 and BRCA2 mRNA expression in sporadic breast carcinomas and its relationship with clinicopathological characteristics. AB - BRCA1 and BRCA2 mRNA expression in sporadic breast cancers was quantified by a real-time reverse transcriptase-polymerase chain reaction (RT-PCR), and the relationship of their expression with various clinicopathological factors was studied. BRCA2 mRNA levels (0.993 +/- 1.395, mean +/- SD (BRCA2 / beta glucuronidase mRNA ratios)) were significantly (P < 0.01) higher than BRCA1 mRNA levels (0.519 +/- 0.570 (BRCA1 / beta-glucuronidase mRNA ratios)), and a weak but significant (r = 0.390, P < 0.01) correlation was observed between BRCA1 and BRCA2 mRNA expression levels. There was no significant association between BRCA1 mRNA expression and clinicopathological factors such as menstrual status, tumor size, lymph node status, estrogen and progesterone receptor status, and histological grade. On the other hand, there was a significant association between higher BRCA2 mRNA expression and estrogen receptor (ER) negativity (P < 0.01) or progesterone receptor (PR) negativity (P < 0.01) or high histological grade (P < 0.01). These results suggest a differential contribution of BRCA1 and BRCA2 in the pathogenesis of sporadic breast cancers. BRCA2 mRNA is speculated to be up-regulated in response to proliferation and genomic instability in high histological grade tumors. PMID- 11429051 TI - Expression of apoptosis regulatory proteins in the skeletal muscle of tumor bearing rabbits. AB - We reported finding that apoptosis occurred in skeletal muscle in the early stage after implantation. In the present study, we investigated expression of the apoptosis-related proteins Bax and Bcl-2 to determine the mechanism of the apoptosis. In the early stage of tumor bearing, 20 days after implantation, lean body mass (LBM) was reduced by 5.06 +/- 1.10% in the tumor-bearing group, compared with an increase of 4.96 +/- 1.26% in the control group. The apoptotic index (AI) of the skeletal muscle in the tumor-bearing group increased to 40.5 +/ 3.20% but was 0% in the control group, and Bax expression was strongly positive in 5 of the 10 rabbits in the tumor-bearing group, and significantly stronger than in the control group (P = 0.0002). In the late stage of tumor bearing, 40 days after implantation, the AI had declined to 0.93 +/- 0.96% in the tumor bearing group, but was still 0% in the control group. Bax expression was rarely detected in either the tumor-bearing group or the control group, and there was no significant difference between the two groups (P = 0.706). No significant changes in Bcl-2 were observed in either group. The above results showed that apoptosis via Bax played a role in muscle wasting associated with progression of the malignant tumor. However, the apoptosis and expression of Bax were seen only in the early stage, within 20 days after implantation, not in the late stage. This suggested that the muscle wasting in the early stage might be caused by a different mechanism from that in the late stage. PMID- 11429052 TI - Ectopic expression of MAFB gene in human myeloma cells carrying (14;20)(q32;q11) chromosomal translocations. AB - Chromosome 14q +, which represents a chromosomal rearrangement involving the immunoglobulin heavy chain gene (IgH) locus, is a genetic hallmark of human multiple myeloma (MM). Here, we report the identification of (14;20)(q32;q11) chromosomal translocations found in MM cells. Double color fluorescence in situ hybridization analyses pinpointed the breakpoints at the 20q11 locus in two MM cell lines within a length of at most 680 kb between the KIAA0823 and MAFB gene loci. Among the transcribed sequences in the vicinity of the breakpoints, an ectopic expression of the MAFB gene, which is located at 450 - 680 kb telomeric to one of the breakpoints and encodes a member of the MAF family basic region / leucine zipper transcription factor, was demonstrated to be associated with t(14;20). This finding, together with that of a previous study describing its transforming activity, suggests that the MAFB gene may be one of the targets deregulated by regulatory elements of the IgH gene as a result of t(14;20). PMID- 11429053 TI - Large-scale analysis of mutations in RET exon 16 in sporadic medullary thyroid carcinomas in Japan. AB - Germline mutations in the RET proto-oncogene are the cause of multiple endocrine neoplasia type 2 (MEN 2A and 2B) and familial medullary thyroid carcinoma (FMTC). Some cases of sporadic medullary thyroid carcinoma (MTC) have also been reported to have mutations in the RET gene. However, two previous reports have given discrepant results on the frequency of the mutations in RET in sporadic MTCs in Japan. To clarify this problem, we analyzed mutations in RET exon 16 in 72 sporadic MTCs by means of the two methods used in the previous studies, direct sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Mutations in exon 16 were detected in only 2 of 72 cases of sporadic MTC. These results suggest that when a MTC has a mutation in RET exon 16, it is more likely to be a hereditary MTC than a sporadic one in Japan. PMID- 11429054 TI - A murine osteosarcoma cell line with a potential to develop ossification upon transplantation. AB - An osteosarcoma cell line has been established from a soft tissue tumor that occurred spontaneously in a BALB / c mouse. This cell line showed ossification when transplanted into syngeneic mice. To examine the mechanism of bone formation, the expression of mRNAs for osteoblastic and chondroblastic markers and factors associated with ossification has been investigated. In culture, the cells exhibited a spindle shape in the growth phase, but had a polygonal shape in the stationary phase. Reverse transcription-polymerase chain reaction analysis showed that the cells expressed mRNAs for pro-alpha1(I) chain of type I collagen, alkaline phosphatase, osteopontin, osteocalcin, and core binding factor alpha1, suggesting differentiation into the stage of osteoblasts during the stationary phase. After transplantation, histological examination revealed small foci of pale blue material and basophilic networks that were scattered in the tumor tissues at one week. The former stained positive with alcian blue, suggesting a chondroid matrix. Pro-alpha1(II) chain of type II collagen mRNA was expressed at one week. A large part of tumors at two and three weeks consisted of basophilic networks, which stained positive via von Kossa's method, indicating a calcified woven bone. In situ hybridization analysis showed strong expression of osteopontin and osteocalcin mRNAs in tumor cells surrounding the bone matrix. Bone morphogenetic protein-6 and -7 mRNAs were detected in transplanted tumors, but not in cultured cells. These results suggest that the cell line has the properties of an osteoblastic lineage when cultured in vitro and has an ossifying ability through endochondral bone formation processes when transplanted in vivo. PMID- 11429055 TI - A novel method for detecting single glandular intestinal metaplasia in the mucosal surface of the fixed stomach using methylene blue. AB - A close association between intestinal metaplasia of the stomach and the well differentiated type of gastric cancer is well recognized. The etiological relationship and how intestinal metaplasia contributes to gastric carcinogenesis are, however, still unclear. In order to answer this question, precise mapping and identification of the smallest lesion of intestinal metaplasia are desired. Establishment of an accurate and easy method for detecting intestinal metaplasia was the goal of this study. Surgical specimens of stomachs resected for gastric cancer were used. The specimens were stained with methylene blue, an oxidation reduction marker, in whole mount, after fixation with 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4), and observed under a stereo-microscope. Normal gastric mucosa was stained blue, whereas intestinal metaplasia mucosa was not stained and had white or sky-blue island-like features. Intestinal metaplasia of complete type was unstained and showed white island-like features, while intestinal metaplasia of incomplete type showed sky-blue staining. With this method, we were able to detect even intestinal metaplasia composed of a single gland, when the intestinal metaplasia was of complete type. When stomach samples were stained in the presence of diphenyleneiodonium chloride (DPI), an inhibitor of nicotineamide adenine dinucleotide phosphate reduced form (NADPH) reductase, all the samples were homogeneously stained blue. Loss of the color of methylene blue was caused by the reductase activity of NADPH reductase, which is strongly and specifically expressed in intestinal metaplasia. A novel method for detecting intestinal metaplasia, even a single gland, was established. PMID- 11429056 TI - Synergistic effects of highly unsaturated fatty acid-containing phosphatidyl ethanolamine on differentiation of human leukemia HL-60 cells by dibutyryl cyclic adenosine monophosphate. AB - Highly unsaturated fatty acid-containing phospholipid (HUFA-PL) has many nutritional and medical applications. We investigated the effect of HUFA-PL on differentiation of human leukemia HL-60 cells induced by dibutyryl cyclic adenosine monophosphate (dbcAMP). HUFA-containing phosphatidylethanolamine (HUFA PE), such as salmon testis PE, significantly enhanced dbcAMP-induced cell differentiation. A combined treatment of 200 mM dbcAMP with 50 mM HUFA-PE increased the nitroblue tetrazolium (NBT)-reducing activity, which is an indicator of differentiation, to a level comparable to that in the case of 500 mM dbcAMP treatment. In contrast, HUFA-lyso PE (a monoacyl form) did not exert an enhancing effect on dbcAMP-induced differentiation. The enhancing effect of HUFA PE was suppressed by a protein kinase C inhibitor, staurosporine, while a protein kinase A inhibitor, H-8, did not suppress the enhancing effect. These findings suggest that HUFA-PE might enhance dbcAMP-induced differentiation through modulation of the protein kinase C signaling pathway in HL-60 cells. PMID- 11429057 TI - Sequential histopathological changes in vivo after suicide gene therapy of gastric cancer induced by N-methyl-N'-nitro-N-nitrosoguanidine in rats. AB - Gastrointestinal cancer is the most important clinical target of gene therapy. Suicide gene therapy, such as with the herpes simplex virus type 1 thymidine kinase (HSV-TK) gene, has been shown to exert antitumor efficacy in various cancer models in vitro. We previously reported in situ gene transfer and gene therapy for gastric cancer induced by N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG) in dogs. Here, we describe the sequential histopathological changes after suicide gene therapy of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced gastric cancer in rats. Gastric tumors were induced by MNNG in 38 / 73 (52%) of Wistar strain rats. The suicide gene therapy group (14 rats) was subjected to in situ gene transfer with a recombinant adenovirus vector carrying the HSV-TK gene driven by CAG promoter (Ad.CAGHSV-TK) in gastric tumor, followed by the antiviral drug ganciclovir (GCV). To observe the histopathological changes at various times after HSV-TK / GCV gene therapy, groups of animals were sacrificed at 3, 8, and 30 days after gene transfer. Apoptosis in the gastric tumors was detected by the TUNEL method to assess the efficacy of HSV-TK / GCV gene therapy, and it was marked in the 8- and 30-day treatment groups compared to the sham operation controls (P < 0.001). Various histopathological changes, degeneration of cancer tissue and fibrosis after necrosis and apoptosis were significantly greater in the 30-day treatment group. The HSV-TK gene was detectable in peripheral blood by PCR until 30 days after gene transfer. These results may be useful in devising a method of suicide gene therapy for humans. PMID- 11429058 TI - Coordinate involvement of cell cycle arrest and apoptosis strengthen the effect of FTY720. AB - A novel reagent, FTY720 (2-amino-2-[2-(4-octylphenyl)ethyl]-1,3-propanediol hydrochloride), has been shown to induce a significant decrease of lymphocytes and lymphoma cells and is expected to be a potent immunosuppressant and anti tumor drug. The decrease in lymphocytes and lymphoma cells is mainly the result of FTY720-induced apoptosis. FTY720 directly affects mitochondria and induces cell death. Moreover, FTY720 activates protein phosphatase (PP) 2A and affects anti-apoptotic intracellular signal transduction proteins to attenuate the anti apoptotic effect. In this study, we examined the relationship between FTY720 induced apoptosis and cell cycle regulation. FTY720 induced apoptosis significantly at the G0 / G1 phase and caused G0 / G1 cell cycle arrest of the human lymphoma cell lines HL-60RG and Jurkat. Simultaneously, retinoblastoma protein (pRB) was dephosphorylated, suggesting that dephosphorylation of pRB was related to FTY720-induced G0 / G1 cell cycle arrest. Because this dephosphorylation was completely blocked by a specific PP1 / 2A inhibitor, okadaic acid, it appears that FTY720-activated PP2A is essential for FTY720 induced cell cycle arrest. FTY720-induced apoptosis was inhibited by Bcl-2 overexpression in Jurkat cells, but this did not prevent FTY720-induced cell cycle arrest, suggesting that the mechanism of FTY720-induced cell cycle arrest is independent of the mechanism of FTY720-induced apoptosis. These two independent pathways strengthen the effect of FTY720. PMID- 11429059 TI - Effect of combination therapy with matrix metalloproteinase inhibitor MMI-166 and mitomycin C on the growth and liver metastasis of human colon cancer. AB - Several synthetic inhibitors of matrix metalloproteinases (MMPs) show antitumor, antimetastasis and antiangiogenesis effects in various models. Synergistic effects of combinations with conventional cytotoxic agents were reported previously. In this study, we examined the effects of a new selective MMP inhibitor, MMI-166, on tumor growth, angiogenesis and metastasis in a liver metastatic model of human xenotransplanted colon cancer (TK-4). We also investigated the synergistic effects of MMI-166 and a conventional cytotoxic agent, mitomycin C (MMC), in this model. Mice transplanted orthotopically with TK 4 were divided into 4 groups; a control group (treated with vehicle solution), an MMI-166 group in which MMI-166 was orally administered (p.o.) at a dose of 200 mg / kg, 6 days / week for 5 weeks, an MMC group in which MMC was administered intraperitoneally (i.p.) at a dose of 2 mg / kg / week for 5 weeks, and a combination group (treated with MMI-166 and MMC). MMI-166 did not inhibit transplanted tumor growth, but significantly inhibited liver metastasis compared with the control group and MMC group (P < 0.01). Significant antitumor and antimetastatic effects of the combination therapy were demonstrated. The microvessel density (MVD) detected by immunohistochemical staining with ER-MP12 antibody tended to be lower in the MMI-166 and the combination groups. These results suggest that MMI-166 has potential antimetastatic ability and a synergistic effect with MMC. PMID- 11429060 TI - Screening of differentially expressed genes in 5-fluorouracil-resistant human gastrointestinal tumor cells. AB - To identify genes differentially expressed in association with resistance to 5 fluorouracil (5FU), an mRNA differential display (DD) analysis was used to compare transcripts from the NUGC-3 human gastric tumor cell line and the NUGC-3 / 5FU / L line, which had acquired 208-fold resistance as a consequence of repeated exposure to escalating concentrations of 5FU. The 110 cDNA fragments differentially expressed in the DD analysis of either the NUGC-3 or NUGC-3 / 5FU / L cells were sequenced and subjected to a homology search, and 29 overexpressed and 22 underexpressed genes were identified in NUGC-3 / 5FU / L as a result. To confirm whether the changes in the gene expression levels in the NUGC-3 / 5FU / L cells were shared by other 5FU-resistant cells, 35 genes were analyzed by northern hybridization in 3 pairs of parent / 5FU-resistant human gastrointestinal tumor cell lines. The analysis revealed 20 overexpressed and 10 underexpressed genes in at least one of the three 5FU-resistant cells as compared with those in the parent cells. Among them, P-glycoprotein, equilibrative nucleoside transporter 1, and methylenetetrahydrofolate dehydrogenase were highly expressed in two of the three 5FU-resistant cells and cytidine deaminase and integrin alpha3 were underexpressed. The acquisition of resistance to 5FU by tumor cells may result from multiple changes in cellular functions. PMID- 11429061 TI - Treatable subsets in cancer of unknown primary origin. AB - The purpose of this study was to investigate the treatable subsets in cancer of unknown primary origin (CUP). Fifty patients (27 males and 23 females; median age, 53 years) with CUP diagnosed between April 1992 and June 1999 were analyzed retrospectively. Of the 50 patients, 39 received chemotherapy: platinum-based in 31, non-platinum-based in 4, and clinical trials of new agents in 4. Of the 39 patients, 13 (33.3%; 95% confidence interval: 19.1 - 50.2%) showed objective responses, with 4 complete responders. Patients with poorly differentiated carcinomas in whom beta-subunit of human chorionic gonadotropin (beta-HCG) was elevated more than 10 mIU / ml and female patients with peritoneal adenocarcinomatosis achieved high response rates (83.3% and 80%, respectively) with platinum-based chemotherapy, as compared with only a 15.3% response rate in the remaining patients. Platinum-based chemotherapy provided promising results in patients with poorly differentiated carcinomas and in female patients with peritoneal adenocarcinomatosis. Significantly elevated serum levels of beta-HCG in patients with poorly differentiated carcinoma might predict a better response to platinum-based chemotherapy. However, the investigation of novel chemotherapeutic approaches is warranted for other groups of patients with CUP. PMID- 11429063 TI - What is the value of ENT specialist outreach clinics? AB - Ear, nose and throat (ENT) specialist outreach clinics, in which hospital-based consultants hold clinics in general practice surgeries, have been popular with general practitioners (GPs) and patients. This prospective study recorded data on 1155 consecutive patients seen by one ENT surgeon in two GP surgeries. At each consultation, a record was kept of the requirement for further investigations that would normally be done at the same time as the consultation in a hospital department. The results showed that 76 per cent of patients needed an investigation, which would be readily available in a hospital but not in a GP surgery (audiometry, endoscopy, microscopy of the ear, a minor procedure or X ray). This study indicates that despite the apparent convenience of outreach ENT clinics to patients and GPs, patients may need to spend more time being assessed than they would if they were investigated in one visit to a hospital department. Unless an outreach clinic is used frequently, it is difficult to justify the cost of equipping it to the same level as a hospital department. Limited resources would be better spent providing good access to well-equipped regularly-used hospital ENT outpatient departments. PMID- 11429062 TI - X-irradiation induces up-regulation of ATM gene expression in wild-type lymphoblastoid cell lines, but not in their heterozygous or homozygous ataxia telangiectasia counterparts. AB - Ataxia-telangiectasia (AT) is an autosomal recessive disease. The relevant gene has been cloned and designated ATM. We studied the expression of both ATM mRNA and the ATM protein in unirradiated and X-irradiated EBV (Epstein-Barr virus) transformed lymphoblastoid cell lines (LCLs) derived from donors who were normal (ATM + / + ), AT heterozygotes (ATM + / - ), or AT homozygotes (ATM - / - ), respectively. In ATM + / + LCLs, the levels of ATM mRNA were found to have increased by approximately 1.5-fold within 1 h of exposure to 10 Gy of X-rays, while the ATM protein levels had increased by 1.5- to 2.0-fold within 2 to 3 h of irradiation. The wild-type mRNA and protein levels both returned to their basal values fairly quickly after this time. The results obtained with the ATM + / - LCLs were quite different, however: neither the mRNA nor protein levels were found to have increased as a consequence of X-irradiation in any ATM + / - LCL. Twelve of the mutations in the ATM - / - LCLs we used were truncating mutations, and we suspected that the corresponding truncated ATM proteins would be too labile to be detected by western blot analysis. However, five of the ATM - / - LCLs produced mutant ATM proteins that were identical in molecular weight to the wild-type ATM protein. When cells from three of these five clones were exposed to X-rays, transcription of the mutant ATM genes appeared to reduce somewhat, as were the levels of protein being produced. These results suggest that the normal ATM gene responds to ionizing radiation by up-regulating its activity, whereas none of the mutant ATM genes we studied were able to respond in this way. PMID- 11429064 TI - Stapedectomy following tympanoplasty. AB - The aim of this study was to evaluate the success of stapedectomy in patients who have previously had a tympanoplasty because of chronic otitis media (COM). Fourteen patients from a private otology practice had undergone tympanoplasty for COM and subsequently underwent stapedectomy. Measurements were taken of the air bone gap (ABG) closure and pure tone average (PTA) which showed hearing improvement. Patients had a mean 36.9 dB PTA hearing gain with 79 per cent closing the ABG to within 20 dB. The need for stapedectomy alone is a rare occurrence for patients with a history of COM requiring a tympanoplasty. Hearing improvement following stapedectomy in these cases was significant, although somewhat less than following traditional stapedectomy in otosclerosis alone. PMID- 11429065 TI - Morphometric analysis of anatomical relationships of the facial nerve for mastoid surgery. AB - Surgical anatomical relationships of the facial nerve (FN) with several landmarks used in mastoid surgery were studied in temporal bone axial high resolution CT scans of 90 patients (180 ears). The shortest distances between the FN and external auditory canal (EAC), sigmoid sinus (SS), posterior fossa dural plate (PFD), and joint of the bony EAC with the lateral surface of the mastoid (M) were measured. These measurements were also analysed in respect of pneumatization and side differences. On average, it was found that FN-EAC was 2.9 mm, FN-SS was 10.5 mm, FN-PFD was 7.3 mm and FN-M was 15.3 mm. FN-EAC was found to be longer in poorly pneumatized bones whereas other distances were longer in pneumatized bones. FN-M was found to be longer on the right side. PMID- 11429066 TI - Quality of life in laryngectomees: a post-treatment comparison of laryngectomy alone versus combined therapy. AB - Patients with advanced and recurrent laryngeal and hypopharyngeal cancer often need combined therapy, which is associated with greater morbidity than single modality treatment. The aim of this study was to assess the quality of life in laryngectomees and to assess whether differences exist between the irradiated and the non-irradiated patients. The University of Michigan Head and Neck Quality of Life questionnaire, a validated instrument for assessing the head and neck cancer related functional status and well-being, was used for this purpose. Subjects for the study included patients who had undergone total laryngectomy for laryngeal or hypophyarngeal squamous cancer. No significant difference between the groups was evident in the various domain scores, although a trend towards higher scores was seen in the combined therapy group. This study suggests that long-term side effects induced by radiotherapy do not adversely affect the quality of life in laryngectomised patients. PMID- 11429067 TI - Blood splash and tonsillectomy: an underestimated hazard to the otolaryngologist. AB - Mucocutaneous exposure is an important route of transmission of hepatitis viruses and HIV in healthcare workers. Few data exist in the literature on the risk of transconjunctival exposure during many surgical procedures. We investigated the use of eye protection during tonsillectomy and measured the incidence of potential eye splash. No otolaryngologist surveyed routinely used eye protection during tonsillectomy. Splash events occurred in 23 out of 103 tonsillectomies. In all cases the surgeon was unaware of the splash. Splash events were significantly more likely to occur during adult tonsillectomy (p < 0.05). We strongly advise the routine use of eye protection during tonsillectomy. PMID- 11429068 TI - Classification of haemorrhage following tonsillectomy. AB - To date there exists no sensible way of classifying the intensity of haemorrhage following tonsillectomy, though this is a prerequisite when comparing the results presented in literature. We evaluated the incidence of post-operative haemorrhage according to our classification in 602 patients who underwent tonsillectomy at our department in 1999. In 21 patients a grade 1 (spontaneous cessation) bleeding occurred. One patient had a grade 2 (infiltration anaesthesia), 14 patients had a grade 3 bleeding (treatment under general anaesthesia) with one patient receiving a blood transfusion. Two patients had a grade 4 bleeding (ligature of the external carotid artery). There was no patient with grade 5 bleeding (lethal outcome). Grades 1 and 3 had the same incidence rates. Primary haemorrhage (<24 hours) can be expected in the majority of patients undergoing tonsillectomy. Secondary haemorrhage is rare and can be life-threatening. PMID- 11429069 TI - Long-term clinico-radiological assessment of endoscopic stapling of pharyngeal pouch: a series of cases. AB - The endoscopic division of the pharyngeal pouch wall with a mechanical stapling device has become increasingly popular. When compared to open excision, the reduced operative time, early resumption of oral intake, and short in-patient stay with no early recurrence of symptoms, are the proposed advantages. Small pouches or thick walled pouches are not suitable for stapling. From December 1996 to December 1999, 32 patients were admitted to the Aberdeen Royal Infirmary for endoscopic stapling of a pharyngeal pouch. Five patients were unsuitable for stapling. In addition, three patients were treated for pouch recurrence after an external approach. Two patients required repeat stapling at a later date. Our results are encouraging in terms of short operation time and hospital admission, improvement of symptoms and minimal complication rate. Fifteen patients were assessed 24 months after the procedure with satisfaction surveys and contrast swallow X-rays. Subjective improvement was sustained throughout this period, despite radiological evidence of persistence of the pharyngeal pouch. PMID- 11429070 TI - Cervical paragangliomas: diagnosis, management and complications. AB - Sixteen patients were diagnosed as suffering from cervical paragangliomas. Eleven patients (68.75 per cent) had twelve carotid paragangliomas (CPs), and five patients (31.25 per cent) had six vagal paragangliomas (VP). One CP (8.33 per cent) originated from paraganglia around the common carotid artery (CCA). Three cases of multiple paragangliomas are presented (18.75 per cent). In 80 per cent (4/5) of VP patients there was widening of the carotid bifurcation similar to that seen with CP. This widening occurred whenever the VP was large enough to grown in between the external carotid artery and internal carotid artery (ECA and ICA). Large VPs may displace the vessels either anterolaterally or anteromedially. Knowledge of the direction of the carotid displacement is essential to avoid intra-operative vascular injuries. Colour flow doppler ultrasound (CFD-US) was found to be a good non-invasive method for diagnosis of vascular neck swellings. It enabled the diagnosis of CP with 100 per cent accuracy, but it was not sufficient for diagnosis of high VP. A transcervical approach, cutting the digastric muscle and the styloid process with the attached ligaments and muscles, was sufficient for excision of most VP. However, midline mandibulotomy might be necessary with high VP. Vascular injuries occurred in 12.5 per cent (2/16) of patients. Superior laryngeal nerve and hypoglossal nerve paralysis occurred, respectively, in (2/11) and (1/11) of patients with CP. Vagal paralysis occurred in all patients with VP. Cerebrovascular accident and post operative death occurred in one patient (6.26 per cent). PMID- 11429071 TI - Management of paediatric otitis media with effusion in the UK: a survey conducted with the guidance of the Clinical Effectiveness Unit at the Royal College of Surgeons of England. AB - A survey was undertaken of the practice of all Otorhinolaryngologists in England and Wales looking both at practice in general and at management of individual patients, including a patient satisfaction questionnaire, under the auspices of the Clinical Effectiveness Unit at the Royal College of Surgeons. Postal questionnaires enquired about practice in general and management in particular of 20 sequential patients per consultant. Parent questionnaires were sent six weeks and one year post-operatively. Ninety consultants took part and 1503 individual patient proformas were returned. Results showed that these consultants are managing children with otitis media with effusion (OME) in a manner very close to the recommendations of the Effective Healthcare Bulletin (1992) and consistent with the recent Clinical Effectiveness Statement of the BAOHNS, and that the intervention is popular with parents. PMID- 11429072 TI - The 'double bag': a device for protection against blood spray in the management of epistaxis. AB - Nasal bleeding is one of the commonest emergencies that place medical staff at risk of blood contamination during management. A sizeable proportion of incidents have been reported where blood splashes in the facial area caused such contamination, and the exposed conjunctivae are potential sites for transmission of human immunodeficiency viruses (HIV). We present a simple self-made device named the 'Double Bag', which is highly effective in preventing blood spraying during the management of epistaxis. It was designed to be used while treating epistaxis or removing nasal packing after surgery, when transient bleeding is expected. PMID- 11429073 TI - Relapsing polychondritis associated with monoclonal gammopathy in a patient with myelodysplastic syndrome. AB - Relapsing polychondritis is a rare condition characterized by inflammation and subsequent degeneration of cartilages. Deformity of the pinna, nasal saddling and stridor due to involvement of the cartilages of the respiratory tract may lead to patients being referred to the otolaryngologist for initial assessment and further management. Recent observations have suggested that relapsing polychondritis may occur as a paraneoplastic phenomenon in cases of myelodysplasia. The case of a patient with relapsing polychondritis, myelodysplastic syndrome and a monoclonal gammopathy is presented. The authors highlight the apparent existence of this association and encourage otolaryngologists to consider such possible links when cases of relapsing polychondritis present to the outpatients department. PMID- 11429074 TI - Artificial tympanic membrane and ossiculoplasty. AB - We report an unusual case of a subjective hearing improvement, confirmed by audiometry, in a patient using a paper towel prosthesis. PMID- 11429075 TI - Extensive metastatic renal cell carcinoma presenting as facial nerve palsy. AB - Metastatic lesions of the parotid gland are well described in the literature. Metastatic spread to the parotid from renal cell carcinoma is rare. We present the only reported case of facial nerve palsy caused by a metastasis to the parotid from a renal cell carcinoma. PMID- 11429076 TI - Tuberculous meningitis presenting as sensorineural hearing loss. AB - We report a 60-year-old male who presented to the Otorhinolaryngology department with an acute unilateral sensorineural hearing loss associated with fever and night sweats. The diagnosis of tuberculous meningitis was made. Unilateral sensorineural hearing loss as a presenting symptom of tuberculous meningitis has not been previously reported. PMID- 11429077 TI - Rhinophycomycosis. AB - Rhinophycomycosis, a subcutaneous fungal infection, is less uncommon than many other frequently cited granulomatous lesions of the paranasal sinuses. Four illustrative cases are presented with their characteristic clinical, radiological and histopathological features. The initial origin of the swelling was always in the region of the nasal vestibule and ala. The therapeutic response to potassium iodide, ketaconazole and fluconazole was dramatic and sustained. PMID- 11429078 TI - Pott's puffy tumour: an unusual presentation and management. AB - Pott's puffy tumour is a rare clinical entity in this era of antibiotics. It is usually seen as a complication of frontal sinusitis. This is the first report of Pott's puffy tumour presenting as a complication of maxillary sinusitis. This is also the first reported case of Pott's puffy tumour treated with debridement and gentamicin beads. We discuss the clinical presentation and successful treatment of this rare disease. PMID- 11429079 TI - External carotid artery aneurysm in an infant presenting with oropharyngeal haemorrhage. AB - We report an unusual case of a mycotic external carotid artery aneurysm presenting in a nine-month-old infant. She presented with coryzal-like symptoms and oropharyngeal haemorrhage. This was thought initially to be due to acute tonsillitis. There was significant haemorrhage and reversible hypovolaemic arrest. A contrast computerised tomography scan (CT) confirmed the diagnosis of a mycotic external carotid artery aneurysm. This was subsequently successfully treated by percutaneous embolization with microcoil insertion. PMID- 11429080 TI - Inflammatory pseudotumour in the submandibular region. AB - We present a rare case of inflammatory pseudotumour in the submandibular region. A review of the literature revealed that this is only the second case of inflammatory pseudotumour in the submandibular region to be reported. Clinical presentation and management of this condition are discussed. PMID- 11429081 TI - A very rare benign tumour in the parotid region: calcium pyrophosphate dihydrate crystal deposition disease. AB - Calcium pyrophosphate dihydrate crystal deposition disease, exhibits several clinical manifestations, from absence of symptoms to severely destructive arthropathy or conditions simulating neoplasm, which is frequently related to the temporomandibular joint. Fifteen of the 31 reported cases of tophaceous pseudogout were found in the head and neck region. A patient presented with a parotid swelling, which initially was suspected to be malignant because of the following findings: radiodensity, progression into the joint, osseous destruction of the major ala of the sphenoid and a fine needle aspirate with crystals, osteoblasts, megakaryocytes and irregular cells of varying size. At surgery there was found a tumour consisting of a white, firm gritty material. It progressed to the skull base where material had to be left, because of the presence of the nerves and vessels. A frozen specimen was reported to be benign. Histological examination showed inflammatory cells, macrophages, a chondroid material with embedded metaplastic chondroid cells and giant cells of foreign body type. Crystal examination of X-ray diffraction revealed calcium pyrophosphate dihydrate. PMID- 11429082 TI - Chondrosarcoma of the thyroid cartilage. AB - Sarcomas of the larynx are extremely rare neoplasms that account for approximately one per cent of all tumours of this organ. We present a case of laryngeal chondrosarcoma originating from thyroid cartilage, characterized by an unusual long clinical course over 15 years without laryngeal symptoms or duplication of metastases, treated at the Suleyman Demirel University Medical Faculty, Ear, Nose and Throat (ENT) department. PMID- 11429083 TI - The crucial role of imaging in detection of facial nerve haemangiomas. AB - Facial nerve haemangioma is a rare benign neoplasm accounting for 0.7 per cent of all tumours involving the temporal bone. The diagnosis of a facial nerve tumour is often missed or delayed. Early diagnosis is imperative as it influences the eventual outcome for facial nerve function. Prognosis is related to the size of the tumour, the severity and the duration of pre-operative paralysis. The definitive diagnosis of a facial nerve tumour rests exclusively with high resolution imaging of the temporal bone using enhanced magnetic resonance imaging (MRI) and thin-sectioned computed tomography (CT). This case emphasizes the crucial role that high quality imaging can play in the diagnosis of facial nerve tumours, and elegantly illustrates the imaging features of facial nerve haemangiomas. PMID- 11429084 TI - Calcium oxalate granuloma of the nose of a chronically dialysed nephritic patient. AB - The patient was a 54-year-old woman who had been suffering from chronic tubulo interstitial nephritis for about seven years, requiring haemodialysis. More recently, she developed a polypoid mass in the left nasal cavity causing discomfort on breathing and slight epistaxis. The tumour was of gritty consistency and measured 28 x 8 x 5 mm. Microscopy showed a lobulated almost cystic structure composed of granulation tissue with comparatively few plasma cells and many multinucleated giant cells lining the spaces filled with crystalline deposits of calcium oxalate. PMID- 11429085 TI - Depression and traumatic brain injury. PMID- 11429086 TI - The prevalence and symptom rates of depression after traumatic brain injury: a comprehensive examination. AB - PRIMARY OBJECTIVE: Differing definitions of depression, limited sample sizes, and variability in methodologies have contributed to equivocal findings about the prevalence of depression among persons with traumatic brain injury. The present investigation used standardized diagnostic criteria and a large sample to identify the manifestations of depression after TBI. METHODS AND PROCEDURES: 722 outpatients with brain injury, referred for comprehensive assessment at a regional Level I trauma centre, were studied. Depressive symptoms were characterized utilizing standard DSM-IV criteria and the Neurobehavioural Functioning Inventory. RESULTS: Forty-two per cent of patients with brain injury met the prerequisite number of symptoms for a DSM-IV diagnosis of major depressive disorder. Fatigue (46%), frustration (41%), and poor concentration (38%) were the most commonly cited manifestations of depression. CONCLUSIONS: Many patients with brain injury are at great risk for developing depressive disorders. Future research should focus on prognostic factors, developing protocols for identification of high risk patients, and examining the efficacy of treatment interventions. PMID- 11429087 TI - A computer system for continuous long-term recording, processing, and analysis of physiological data of brain injured patients in ICU settings. AB - The objective of this project was to assemble and test a computer-based system for continuous long-term physiological data acquisition. The system would be used to study short-term (heart rate variability) and long-term (circadian rhythms) dynamics of physiological parameters in severely brain injured patients in ICU settings. A system has been built using open-source software and the Linux operating system as the platform. The system consists of three main parts: data acquisition, processing and analysis. The system was tested in ICU and experimental settings for long periods of time (up to 10 days of non-stop recording). The system appeared to function properly and accurately. Samples of the data according to the stages of acquisition-analysis process are presented in the paper. Avenues for the system use and development are discussed. PMID- 11429088 TI - Unilateral visual neglect in right-hemisphere stroke: a longitudinal study. AB - Unilateral visual neglect occurs frequently after right-hemisphere stroke. This longitudinal study assessed the performance pattern of 54 individuals with unilateral right-hemisphere stroke on the Behavioural Inattention Test (BIT), a measure of unilateral neglect. Eighteen subjects returned for retesting between 6 26 months post-onset. Results confirmed the frequent occurrence of unilateral visual neglect in patients with right-hemisphere damage. A high correlation was found between the Conventional and Behavioural Subtests of the BIT, indicating that the shorter subtest may be sufficient to identify neglect. Longitudinal data revealed two subgroups, those with transient neglect that resolved within 6 months and those with neglect that persisted over time. No differences in aetiology and lesion location distinguished patients with and without neglect or patients with transient or persistent neglect. Further research is needed to confirm these results and identify other characteristics that would predict the nature of the neglect. PMID- 11429089 TI - Cognitive deficits following blast injury-induced neurotrauma: possible involvement of nitric oxide. AB - Blast injuries, that is injuries caused by the complex pressure wave generated by some explosions, show increasing frequency throughout the world. However, whether blast injury is capable of inducing memory dysfunction has not been previously investigated. The present study examines the effects of blast injury-induced neurotrauma on memory deficit in rats. Furthermore, it is hypothesized that blast injury, stimulating nitric oxide production in the medial mesodiencephalic reticular formation and the dorsal hippocampus, both structures being involved in memory processing, may induce memory deficits. Prior to blast injury, Wistar rats were trained for an active avoidance task for 6 days. On day 6, rats that had acquired the avoidance response were subjected to whole-body blast injury, using a BT-I shock tube. Neurotrauma was confirmed by electron microscopical examination. At the completion of cognitive testing, rats were sacrificed at 3, 24 hours and 5 days after injury. The nitric oxide production in the brain structures was determined by the total nitrite/nitrate concentration, and by the expression of inducible nitric oxide synthase mRNA. The rats with blast injury revealed significant deficits in performance of the active avoidance task that persisted up to 5 days post-injury. Electron microscopical findings in both brain structures showed swellings of neurons, glial reaction, myelin debris, and increased pinocytotic activity on the fifth day following trauma. In blast injured rats, there was a significant elevation in total nitrite/nitrate levels 3 and 24 hours following injury which was comparable with the changes in the expression of inducible nitric oxide synthase mRNA. The results indicate that blast injury-induced neurotrauma is able to cause cognitive deficits. PMID- 11429090 TI - Rehabilitation outcomes for brain injured patients in Australia: functional status, length of stay and discharge destination. AB - This study describes the rehabilitation length of stay (LOS), discharge destination and discharge functional status of 149 patients admitted with traumatic brain injury (TBI) to an Australian hospital over a 5-year period. Hospital charts of patients admitted between 1993-1998 were reviewed. Average LOS over the 5-year time period was 61.8 days and only decreased nominally over this time. Longer LOS was predicted by lower admission motor FIM scores and presence of comorbidities. Mean admission and discharge motor FIM scores were 58 and 79, which represented a gain of 21 points. Higher discharge motor FIM scores were predicted by higher admission motor FIM scores and younger age. FIM gain was predicted by cognitive status and age. Most patients, 88%, were discharged back to the community, with 30% changing their living setting or situation. Changing living status was predicted by living alone and having poorer functional status on admission. PMID- 11429091 TI - Combined motor disturbances following severe traumatic brain injury: an integrative long-term treatment approach. AB - Patients surviving severe traumatic brain injury (TBI) often suffer from residual impairments in motor control, communication skills, cognition and social behaviour. These distinctly hamper their capability to return to their 'pre trauma' activity. Comprehensive and integrated rehabilitation programmes initiate, during the acute phase, a prolonged treatment process which starts at the most sophisticated medical systems. There is no clear end point for the treatment of these patients, since the recovery process and the rehabilitation activity may continue for years, even after patients return home to live with their families. The inherent inability to make a firm early prediction regarding outcome of patients and the late appearance of additional symptoms stress the need for a comprehensive close long-term follow-up. The following presentation concerns the description of the treatment strategy and long-term improvement of a 22-year-old male who suffered from very severe TBI. On admission to the emergency room, he was in the decerebrated position and his Glasgow Coma Scale (GCS) was at the lowest (3). The focus of this presentation is on the recovery of motor function. The initial motor disabilities included weakness in all four limbs, in particular left hemiplegia, and right hemiparesis with severe bilateral ataxic elements and a marked tremor of the right arm. Range of motion was limited in hips, and he suffered from stiff trunk and neck. Goals of physiotherapy were directed towards improving range of motion (ROM) and active movement. Casting, use of orthoses, biofeedback, hydrotherapy, hippotherapy, medication and nerve blocks for reducing spasticity were timely applied during the process. The motor improvement in this very severe TBI patient who is now over 3 years post-injury still continues and has a functional meaning. He has succeeded in being able to stand up by himself from a chair and is able to walk unaided and without orthoses for very short distances--up to five steps. He is able to drink soup without assistance and play a few notes on the piano. Marked cognitive improvement occurred as well. It is concluded that motor improvement may be evident over long periods of time and various timely interventions may assist in the process. PMID- 11429092 TI - Conceptual dilemmas in evaluating individuals with severely impaired consciousness. AB - States of severely impaired consciousness (SIC) are characterized by cognitive and motor limitations. This case report describes a 45-year-old female with impaired consciousness who began to 'walk'. She initially presented to the hospital unresponsive and was subsequently diagnosed with metabolic encephalopathy due to severe hypoglycaemia. Traditional indices of consciousness indicated a low level of responsiveness; however, during physical therapy, she displayed reciprocal walking movements when lifted to a standing position by two therapists. Despite her ability to walk increased distances during and after neurorehabilitation, she was unable to consistently demonstrate responses indicative of higher levels of consciousness. This case illustrates the challenge of rating patients with limited behavioural repertoire using established measures of impaired consciousness. PMID- 11429093 TI - Sports-related brain injury research: methodological difficulties associated with ambiguous terminology. AB - A basic premise underlying sound methodological research is that it is based on constructs that have a sound operational definition. Failure to do so impedes comparison with other relevant literature. In sports-related brain injury research, the varied criteria and inconsistent use of the term 'concussion' and its interchangeability with other generic terms such as mild/minor head injury, and mild brain injury exemplifies such a situation. This paper will background the issues involved and provide rationale to support a call for tightening the use of terminology in the area. PMID- 11429095 TI - MAM-6C-8 (Anti-Penicillin Acylase). PMID- 11429094 TI - Regarding the search for a unified definition of mild traumatic brain injury. PMID- 11429096 TI - HA1, HA2 Anti-Human Arginase. PMID- 11429097 TI - ATOM-2, specific for S-peptide-tagged recombinant proteins. PMID- 11429099 TI - ior egf/r3 anti-human EGF receptor. PMID- 11429098 TI - MAb to rat galanin. PMID- 11429100 TI - MAb HT10 (anti-mucin). PMID- 11429106 TI - Fluorescence resonance energy transfer analysis of CCR-V64I and SDF1-3'a polymorphisms: prevalence in southern Spain hiv type 1+ cohort and noninfected population. AB - The relationship between host genotype and AIDS, as well as the different genotype frequencies observed in different populations, have become important topics in HIV research. Therefore, the development of methods that provide faster and reliable results may contribute to further development and knowledge of those topics. We present the results of genotyping SDF1-3'A and CCR2-V64I in 440 HIV-1 infected people and 100 noninfected controls from southern Spain, using a novel method based on real-time PCR with LightCycler technology and fluorescence resonance energy transfer. Frequencies obtained were 23.8% for SDF1-3'A and 9.5% for CCR2-V64I for both HIV+ cohort and general population. Both polymorphisms are in accordance with the Hardy-Weinberg equilibrium law and no differences between patients and controls have been observed. PMID- 11429107 TI - Trial-related discrimination in HIV vaccine clinical trials. AB - Participants in preventive HIV vaccine trials may experience negative social consequences of trial participation, including problems related to a vaccine induced positive HIV antibody test, yet few vaccine studies have reported on this issue. From October 1995 through November 1998, 1516 AIDS Vaccine Evaluation Group (AVEG) volunteers were assessed for reports of trial-related discrimination (TRD). Ninety TRD events were reported by 76 (5%) of 1516 volunteers. The most commonly reported incidents (n = 52, 57.8%) were negative reactions of friends, family, and co-workers to the volunteer. Few incidents (approximately 10%) were reported as linked to HIV testing. The majority of events (n = 47, 52%) were described by volunteers as "resolved" at the time of reporting, 36 (40%) as "not resolved," and for 7 (8%) events volunteers did not report resolution status. Reported incidents were analyzed by logistic regression to determine their association with the volunteer's age, sex, race, sexual orientation, and HIV risk category. There was no association between volunteer characteristics and TRD. Logistic regression and analysis of variance (ANOVA) were used to analyze association of trial sites with the number of TRD events reported. After controlling for site variation in data collection and reporting, no significant differences were found between the sites in terms of the number or type of TRD reported. Fears that TRD would be widespread and severe have not been borne out by this analysis. While the results of this study are reassuring, they should be interpreted with caution, as it is unclear whether these results may be extended to phase III trials enrolling large numbers of individuals at higher risk of HIV acquisition. PMID- 11429108 TI - Evidence of two distinct subsubtypes within the HIV-1 subtype A radiation. AB - Members of HIV-1 group M are responsible for the vast majority of AIDS cases worldwide and have been classified on the basis of their phylogenetic relationships into nine roughly equidistant clades, termed subtypes. Although there are no known phenotypic correlates for these genotypes, the disproportionate spread of certain of these lineages has been taken to indicate that subtype-specific biological differences may exist. The subtype nomenclature thus remains an important molecular epidemiological tool with which to track the course of the group M pandemic. In this study, we have characterized HIV-1 strains described previously as unusual subtype A variants on the basis of partial sequence analysis. Six such strains from Cyprus (CY), South Korea (KR), and the Democratic Republic of Congo (CD) were PCR amplified from infected cell culture or patient PBMC DNA, cloned, and sequences in their entirety (94CY017, 97KR004, 97CDKTB48, and 97CDKP58) or as half genomes (97CDKS10 and 97CDKFE4). Distance and phylogenetic analyses showed that four of these viruses (94CY017, 97CDKTB48, 97CDKFE4, and 97CDKS10) were closely related to each other, but quite divergent from all other HIV-1 strains, except for subtype A viruses, which represented their closest relatives. In phylogenetic trees from gag, pol, env, and nef regions, the four newly characterized HIV-1 strains formed a distinct sister clade to subtype A, which was as closely related to subtype A as subsubtypes F1 and F2 are to each other. According to current nomenclature rules, this defines a subsubtype, which we have tentatively termed A2. The two other viruses, 97KR004 and 97CDKP58, as well as a full-length HIV-1 sequence from the sequence database (ZAM184), were found to represent complex A2/D, A2/G, and A2/C recombinants, respectively. These results indicate that HIV-1 subtype A is composed of two subsubtypes (A1 and A2), both of which appear to have a widespread geographic distribution. The A2 viruses described here represent the first reference reagents for this new group M lineage. PMID- 11429109 TI - Phylogenetic analysis of multiple heterosexual transmission events involving subtype b of HIV type 1. AB - Between 1996 and 1999 thirteen cases of HIV infection were detected in Doncaster, a small town in the north of England (population approximately 250,000). A complex network of shared sexual histories involving local nightclubs linked these cases, with the only known risk factor being heterosexual intercourse. A series of frozen blood samples was collected in 1998-1999 and amplified by PCR to generate full-length gp120 clones. Sequencing demonstrated that all the transmission events in this heterosexual group involved the B subtype of HIV-1. When relationships between the samples were assessed it became clear that these 13 cases represented at least three separate strains of HIV-1, indicating that HIV is well established in this community. Eleven of the 13 cases were related, forming two distinct groups. Further investigation revealed that one group contained five patients whose general health was good and who were not receiving HAART. In contrast, the second group of six patients, including the putative index case, were symptomatic, receiving HAART, and may have been infected with a CXCR-4-utilizing virus. Several of the cases that were linked by genetic criteria were not linked by contact tracing, implying that further undiagnosed cases may exist in this community. To our knowledge, this is the largest outbreak of HIV studied within the heterosexual community in the United Kingdom to date, suggesting that this route of infection is becoming more common within the United Kingdom. PMID- 11429110 TI - CCR5 is the major coreceptor used by HIV-1 subtype C isolates from patients with active tuberculosis. AB - Tuberculosis (TB) is the major opportunistic infection of HIV-infected patients in developing countries and is associated with activation of the immune system and increased HIV-1 expression. The aim of this study was to explore the biological properties of HIV-1 isolates from patients with active TB. Ten HIV-1 subtype C isolates were analyzed for biological phenotypes, using MT-2 cells, and for coreceptor usage, using coreceptor-transfected cell lines. All isolates were nonsyncytium inducing (NSI) in the MT-2 assay and replicated in CCR5-expressing cells. None of the isolates used CXCR4 or any of the minor coreceptors (CCR1, CCR2b, or CCR3) efficiently. Analysis of the V3 region showed that all isolates contained the GPGQ motif characteristic of subtype C and also had a sequence profile typical of NSI viruses. These data indicate that despite their advanced disease state, patients with TB harbor viruses that use the CCR5 coreceptor. It is possible that activation of monocytes and macrophages during TB infection results in the expansion of macrophage-tropic isolates that preferentially use CCR5. PMID- 11429111 TI - An HLA-directed molecular and bioinformatics approach identifies new HLA-A11 HIV 1 subtype E cytotoxic T lymphocyte epitopes in HIV-1-infected Thais. AB - Only limited cytotoxic T lymphocyte (CTL) epitope mapping has been done in nonsubtype B HIV-infected persons. We used molecular immunogenetic tools to determine HIV-specific CTL responses in HIV-1 Env subtype E-infected female sex workers (FSWs) from northern Thailand, where more than 50% of the population is HLA-A11 positive. EpiMatrix, a computer-based T cell epitope prediction algorithm, and a manual editing approach were used to predict 77 possible HLA-A11 CTL epitopes in HIV-1, some of which were conserved between subtypes B and E. MHC binding of these peptides was determined in an HLA-A11 stabilization assay, and binding peptides were tested for CTL recognition in eight HLA-A11-positive FSWs. Subtype E versions of known HLA-A2 subtype B HIV epitopes were also tested in four HLA-A2 positive FSWs. CTL responses were detected in all HLA-A11-positive and in three of four HLA-A2-positive persons. Among the 12 FSWs responses to peptides were found to Pol in 9 (75%), Env in 7 (58%), Nef in 5 (42%), and Gag in 5 (42%), and to conserved epitopes in 8 (67%). To identify HLA-A11 CTL epitopes in the absence of prediction tools, it would have been necessary to test almost 3000 10-mer peptides. EpiMatrix and manual predictions reduced this number to 77, of which 26 were MHC binding and 12 were CTL epitopes. Six of these HLA-A11 CTL epitopes have not been previously reported and are located in RT, gp120, and gp41. This report of CTL responses in subtype E-infected individuals defines epitopes that may be useful in HIV pathogenesis or vaccine studies. PMID- 11429112 TI - HIV-specific cytotoxic T lymphocytes, HLA-A11, and chemokine-related factors may act synergistically to determine HIV resistance in CCR5 delta32-negative female sex workers in Chiang Rai, northern Thailand. AB - Understanding how highly HIV-exposed individuals remain HIV uninfected may be useful for HIV vaccine design and development of new HIV prevention strategies. To elucidate mechanisms associated with resistance to HIV infection, immunologic and genetic factors were examined in 14 HIV-exposed but persistently seronegative (HEPS) female sex workers from Chiang Rai, northern Thailand and in ethnically matched, HIV-positive (n = 9) and HIV-negative women (n = 9). The HEPS women were identified in a study of commercial sex workers who had an HIV-1 incidence of 20.3 per 100 person-years. A high frequency of HLA-A11 was observed in HEPS women (86%) compared with northern Thai controls (56%). HIV-specific cytotoxic T lymphocyte (CTL) lytic responses were detected in cryopreserved peripheral blood mononuclear cells (PBMCs), using HLA-A-matched subtype E HIV-1 peptides in four of seven (57%) HEPS women, eight of eight HIV-positive women, and zero of nine HIV-negative unexposed controls (p = 0.019 HEPS women vs. HIV-negative controls). CTL lysis levels were low, but responses were detected to peptides from Nef, Pol, Gag, and Env. Nef responses predominated in HEPS women. Compared with controls, HEPS women tended to have higher frequencies of CCR5 promotor 59402GG and SDF-1 3'UTR 801A genotypes known to influence HIV transmission or course of disease. HEPS women also had higher levels of spontaneous RANTES production by PBMCs than other groups. Each of these factors could potentially contribute to HIV resistance. As most HEPS women had one or more of these factors, they may prevent HIV infection synergistically by blocking HIV cell entry, delaying its dissemination, or killing HIV-infected cells. PMID- 11429113 TI - Production of a novel viral suppressive activity associated with resistance to infection among female sex workers exposed to HIV type 1. AB - To investigate mechanisms of natural resistance to human immunodeficiency virus type 1 (HIV-1), we obtained blood samples from eight women who remained HIV-1 negative after > 3 years of high-risk sex work in Chiang Rai, Thailand. CD4+ T lymphocytes from these highly exposed, persistently seronegative (HEPS) women were readily infectable in vitro with HIV-1 subtypes B and E. Autologous CD8+ cell suppression of both HIV-1 subtypes was evident in HEPS infection cultures, but to an extent also observed in cultures from non-HIV-exposed individuals. Furthermore, production of beta-chemokines was not enhanced in HEPS cultures. However, HEPS cultures displayed significantly enhanced production of a soluble activity that suppressed postintegrated HIV-1 replication. This activity was the unique product of CD4+ T cell and monocyte cocultures. Therefore, although HEPS individuals are apparently susceptible to infection, the production of a postintegrated HIV-1 suppressive activity during monocyte-T cell interactions might protect against the establishment of infection by limiting viral dissemination. PMID- 11429114 TI - Comparative pathobiology of HIV- and SIV-associated lymphoma. PMID- 11429115 TI - Analysis of HIV type 1 protease and reverse transcriptase sequences from Venezuela for drug resistance-associated mutations and subtype classification: a UNAIDS study. AB - We report the first study on prevalence of antiretroviral drug-associated resistance mutations in Venezuela. Protease and reverse transcriptase (RT) coding regions were analyzed in DNA samples obtained from 100 HIV-1-infected individuals. Primary resistance mutations to RT inhibitors were identified in 26% of patients treated with these drugs. Transmission of HIV-1-resistant strains was detected in a drug-naive patient (3%). Primary resistance mutations to protease inhibitors (PIs) were present in 9% of the 44 PI-treated patients and in 1 PI naive individual. Phylogenetic analysis of these samples has resulted in the most extensive survey, to date, of HIV-1 genetic forms circulating in Venezuela. Ninety-nine samples clustered with subtype B, and 1 individual harbored the first B/F recombinant virus reported in Venezuela, with protease clustering with subtype F and RT with subtype B. In addition, this isolate had a new insertion (Glu-34 duplication) in the protease gene. PMID- 11429116 TI - Emergence of new HIV-1 subtypes other than Subtype C among antenatal women in Lusaka, Zambia. AB - Molecular epidemiology of HIV-1 in Zambia was investigated by direct sequencing of PCR products from samples collected from antenatal attendees in Lusaka, Zambia. One hundred and forty samples were initially screened for HIV, using antibody assays. Thirty-three (23.6%) samples were HIV-1 positive. Sequences of the HIV-1 env gp120 region were obtained from 28 of 33 (85%) HIV-1-positive samples. Twenty-six of the 28 sequences were HIV-1 env subtype C-like as previously reported. However, one HIV-1 env subtype D-like virus and one HIV-1 env subtype G-like virus were identified. This is the first time that these two HIV-1 env subtype viruses have been identified in Zambia, suggesting that more subtypes could be in existence. PMID- 11429117 TI - Characterization of HIV type 1 tat sequences associated with perinatal transmission. AB - Human immunodeficiency virus type 1 (HIV-1) tat exon I sequences were analyzed from six mother-infant pairs after perinatal transmission. The tat open reading frame was maintained in 140 of the 154 clones analyzed, with a 90.9% frequency of intact tat open reading frames. In addition, a low degree of heterogeneity was observed in tat sequences within mothers, within infants, and between epidemiologically linked mother-infant pairs. However, the distances of tat sequences between epidemiologically unlinked individuals were greater than in epidemiologically linked mother-infant pairs. The infant sequences showed amino acid sequence patterns similar to those present in their respective mothers. The functional domains required for Tat function, including amino-terminal, cysteine rich, core and basic regions, which constitute domains for activation and RNA binding, were highly conserved in most of the sequences. Phylogenetic analysis of 154 mother-infant tat sequences showed that they formed distinct clusters for each mother-infant pair and grouped with subtype B sequence. These findings suggest that an intact and functional tat gene is conserved in HIV-1 mother infant isolates that are involved in perinatal transmission. PMID- 11429118 TI - Characterization and phylogenetic analysis of South African HIV-1 subtype C accessory genes. AB - To acquire new knowledge about the genetic diversity and potential impact on vaccine strategies of HIV-1 subtype C in South Africa, we have characterized the vif, vpr, and vpu genes of 15 isolates. Phylogenetic analysis of the genomic fragment encompassing these genes revealed subtype C subclusters, suggesting close relatedness with subtype C strains from other geographic locations and excluded isolation of South African strains. The putative T155 phosphorylation site in the C terminal of Vif was absent in all subtype C sequences. Variation in the predicted amino acid sequences of the three genes further showed strong correlation with other subtype C sequences. PMID- 11429119 TI - Phylogenetic characterization of a new HTLV type 1 from the Ainu in Japan. AB - Human T cell leukemia virus type 1 (HTLV-1) is endemic among three ethnically distinguishable populations in Japan (the Ainu, Ryukyuans, and Wajin), which, together, account for most of the population in Japan. While much is known about the phylogeny of the Ryukyuan and Wajin strains of HTLV-1, only one Ainu strain has been phylogenetically analyzed. We report here a new HTLV-1 strain from the Ainu. The new isolate (U8306), as well as the previously reported isolate, are members of the Cosmopolitan group and further belong to the Transcontinental subgroup. This subgroup also predominates among the Ryukyuans, whereas the Japanese subgroup is the major subgroup among the Wajin. The predominance of subgroup A in the Ainu and Ryukyuans suggests that they share a common origin of HTLV-1. PMID- 11429120 TI - A phase III study of recombinant human interferon gamma to prevent opportunistic infections in advanced HIV disease. AB - The efficacy and safety of recombinant human interferon gamma (rIFN-gamma) in the reduction of opportunistic disease in patients with advanced HIV-1 infection are assessed. A 12-month double-blind, placebo-controlled, multicenter, Phase III trial of rIFN-gamma in HIV-positive patients with CD4 < 100 x 10(9)/liter on stable antiretroviral therapy. Eighty-four patients were allocated treatment on a 1:1 basis to rIFN-gamma or placebo. Patients received rIFN-gamma 0.05 mg/m(2) or 0.9% saline subcutaneously three times weekly for 48 weeks (optional extension to 18 months). The primary end point was the incidence of opportunist infections (CDC categories B/C). Secondary end points included mortality, immunological, and virological parameters. Patients on placebo had a mean of 3.45 opportunist infections (OIs) in the first 48 weeks. Patients treated with rIFN-gamma had a mean of 1.71 OIs (p = 0.04). However, the model showed overdispersion and the inclusion of a dispersion factor raised the p value to 0.13. rIFN-gamma appeared to have a particular effect on the incidence of Candida, herpes simplex, and cytomegalovirus infections. Three-year survival in the rIFN-gamma arm was 28% compared to 18% in the placebo group (not significant). rIFN-gamma-associated side-effects of headache, fatigue, rigors, influenza-like symptoms, depression, myalgia, and granulocytopenia were reversible. There was no evidence for HIV activation. Although not significant, the trend towards decreased opportunistic infections and increased survival warrants consideration of further trials of rIFN-gamma. The study gives additional information on the safety profile of this cytokine. PMID- 11429121 TI - Quantification of HTLV type I and HIV type I DNA load in coinfected patients: HIV type 1 infection does not alter HTLV type I proviral amount in the peripheral blood compartment. AB - Several reports suggest that HTLV-I/HIV coinfection may be associated with an increased risk of HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). In HTLV-I-monoinfected patients, the occurrence of HAM/TSP is associated with high peripheral blood HTLV-I proviral load. Using a real-time quantitative PCR assay, we assessed the proviral DNA load in peripheral blood mononuclear cells (PBMCs) from 15 asymptomatic HTLV-I-monoinfected patients, 15 HTLV-I-monoinfected patients with HAM/TSP, and 25 HTLV-I/HIV-1 coinfected patients, including 4 with HAM/TSP. We also measured HIV-1 proviral DNA load in PBMCs from the coinfected patients. The median HTLV-I proviral loads were 6,800 and 4,100 copies per 10(6) PBMCs in the asymptomatic monoinfected and coinfected groups, and 58,800 and 43,300 copies per 10(6) PBMCs in the monoinfected and coinfected patients with HAM/TSP, respectively. The difference between HTLV-I proviral loads in HAM/TSP and asymptomatic monoinfected patients was statistically significant (p < 0.0001), but there was no difference between the HTLV-I-monoinfected and HTLV-I/HIV-1-coinfected groups. There was no correlation between HTLV-I and HIV-1 proviral load. HTLV-I proviral load did not correlate with the CD4+ T lymphocyte count. Among patients with no HTLV-I disease, the median copy number of HTLV-I per 10(6) circulating CD4+ T cells was 114,000 in the coinfected group and 16,700 in the monoinfected group, but the difference was not significant (p = 0.089). These data do not confirm the hypothesis in which HIV-1 coinfection would increase HTLV-I proviral burden in the PBMCs. However, depletion of the CD4+ T cell subset, the main target of HTLV-I, could be counterbalanced by an up-regulation of HTLV-I replication or by greater resistance of HTLV-I-infected cells to HIV-1-induced destruction. PMID- 11429122 TI - Selection by AZT and rapid replacement in the absence of drugs of HIV type 1 resistant to multiple nucleoside analogs. AB - We studied the intrahost evolution and dynamics of a multidrug-resistant HIV-1, which contains an insertion of two amino acids (aa) and several aa changes within the reverse transcriptase (RT) gene. From an individual receiving intermittent therapy, sequences of 231 full-length molecular clones of HIV-1 RT were obtained from serum-derived viruses at 12 consecutive time points over a period of 6 years, 17 to 20 clones per time point. In the 3.5-year period prior to the first course of therapy, only wild-type (wt) viruses were found. As soon as 6 months after the start of zidovudine (AZT) monotherapy, all viruses contained an insertion of two aa between positions 68 and 69 of the RT and aa changes at positions 67 and 215, a combination conferring resistance to multiple nucleoside analogs. After termination of therapy, the insertion mutants were rapidly and completely replaced by the wt viruses. In turn, the insertion mutants replaced the wt viruses after initiation of therapy with 3TC, d4T, and saquinavir. After termination of triple therapy, the wt viruses completely replaced the mutants within 1 month, which is markedly faster than has been observed earlier for the replacement of AZT-resistant viruses. Fast replacements of the mutant virus populations after termination of therapy indicate gross competitive disadvantage of the insertion mutant in the absence of therapy, which we estimated by using several models. The insertion mutants attained high virus loads, demonstrating that virus load cannot be used as a direct measure of virus fitness. PMID- 11429123 TI - Stimulation of HIV gp120-specific cytolytic T lymphocyte responses in vitro and in vivo using a detoxified pertussis toxin vector. AB - CD8+ cytolytic T lymphocytes (CTL) are almost certainly an important component of a potentially protective immune response to HIV. To test the ability of pertussis toxin (PT) to deliver an HIV-derived major histocompatibility complex (MHC) class I peptide for CTL stimulation, we constructed a fusion of the gp120 P18-I10 CTL epitope with a genetically detoxified derivative of PT (PT9K/129G) and assayed this fusion for its ability to stimulate a gp120-specific CTL response in vitro and in vivo. Antigen-presenting cells incubated with this fusion protein were lysed by P18-I10-specific CTL in vitro and this activity was shown to be MHC class I restricted. The activity was inhibited by brefeldin A but was not inhibited by proteasome inhibitors, possibly because PT undergoes retrograde intracellular transport through the Golgi apparatus to the endoplasmic reticulum and delivers epitopes directly to nascent class I molecules. Mice immunized intraperitoneally with a single dose of the fusion protein without adjuvant raised a strong gp120-specific CTL response in the spleen. This CTL response was dependent on (1) the dose of fusion administered, (2) the fusion of the epitope with the toxin (since coadministration of peptide and toxin gave no response), and (3) the activity of CD8+ cells. These data demonstrate that this detoxified derivative to PT, which is already a component of a licensed vaccine for humans, could represent a useful vaccine vector molecule for stimulation of HIV-specific CTL responses. PMID- 11429125 TI - A conformational C4 peptide polymer vaccine coupled with live recombinant vector priming is immunogenic but does not protect against rectal SIV challenge. AB - The conserved, immunogenic CD4 binding site on the viral envelope is an attractive HIV or SIV vaccine candidate. Polymerization of an 18 amino acid segment derived from the C4 domain of SIV gp120 produced a peptide polymer or "peptomer," having an alpha-helical conformation possibly mimicking a proposed structure of the C4 domain in the unbound native protein. The SIV peptomer and native gp120 were compared as subunit boosts following two adenovirus type 5 host range (Ad5hr)-SIVenv recombinant priming immunizations. Both vaccine regimens successfully elicited SIV-specific CTL responses in five of six immunized macaques. Peptomer-boosted macaques exhibited significantly higher envelope specific T cell proliferative responses than either the gp120-boosted macaques or controls. Peptomer immunization also elicited peptomer and SIV gp120-specific binding antibodies, but only native gp120 boosting elicited SIV neutralizing antibodies. Upon intrarectal challenge with SIVmac32H, all nine macaques became infected. The solely envelope-based vaccine conferred no protection. However, changing the boosting immunogen to the C4 peptomer did not improve protective efficacy in spite of its elicitation of humoral and cellular immune responses, including robust T-helper activity. In spite of the peptomer's strong immunogenicity and potential for induction of broadly protective immune responses, it was not effective as a subunit vaccine. PMID- 11429124 TI - Enhanced avidity maturation of antibody to human immunodeficiency virus envelope: DNA vaccination with gp120-C3d fusion proteins. AB - DNA vaccination can elicit both humoral and cellular immune responses and can confer protection against several pathogens. However, DNA vaccines expressing the envelope (Env) protein of human immunodeficiency virus (HIV) have been relatively ineffective at generating high titer, long-lasting, neutralizing antibodies in a variety of animal models. In this study, we report that fusion of Env and the complement component, C3d, in a DNA vaccine, enhances the titers of antibody to Env. Plasmids were generated that expressed a secreted form of Env (sgp120) from three isolates of HIV and these same forms fused to three tandem copies of the murine homologue of C3d (sgp120-3C3d). Analyses of titers and avidity maturation of the raised antibody indicated that immunizations with each of the sgp120-3C3d expressing DNAs accelerated both the onset and the avidity maturation of antibody to Env. PMID- 11429126 TI - Molecular epidemiology of HIV type 1 subtypes in equatorial guinea. AB - Equatorial Guinea is endemic for HIV-1. This country borders to the north with Cameroon, where different subtypes belonging to group M, as well as group O strains, are circulating simultaneously. To assess the molecular epidemiology of HIV-1 in Equatorial Guinea we analyzed 76 plasma samples collected throughout 1999 from seropositive individuals. Phylogenetic analysis of the gp41 region revealed that 53 were of subtype A, with 64% of these sequences clustering with CRF02_AG reference strains; 11 were of subtype C; 4 were of subtype D; 2 (closely related to subtype F2) were of subtype F; 3 were of subtype G, two of them forming a separate cluster with the recombinant circulating forms CRF06_cpx; 1 was of subtype H; and 2 were unclassifiable. Although subtype A is predominant, the presence of 14% of subtype C is also noteworthy. This work represents the first HIV-1 subtype distribution study in Equatorial Guinea. PMID- 11429127 TI - Genetic diversity and phylogenetic analysis of human immunodeficiency virus type 1 subtypes circulating in French Guiana. AB - We investigated the characterization of different HIV-1 subtypes present in French Guiana by use of three different methods. Serological methods were used for the initial screening, which were then confirmed by the heteroduplex mobility assay (HMA). The V3 env region was subsequently sequenced for phylogenetic analysis, to confirm the subtype of the samples, and to assign a subtype to samples that gave results that were difficult to interpret or discordant by serology or HMA. A total of 221 HIV-1 seropositive samples were typed; 110 of them were confirmed by HMA and 16 were sequenced. Of the 221 samples tested 210 patients (95%) were found to be infected with subtype B, 10 (4.5%) were infected with subtype A, and one patient was infected with subtype F. Phylogenetic analysis demonstrated that the strains from French Guiana were closely related to the subtype A and B subtypes, and that one strain was closely related to an F subtype (100% bootstrap value). Four strains from French Guiana clustered in the subtype A (99% bootstrap value) and the other strains were associated with subtype B (100% bootstrap value). The geographic position of French Guiana suggested that HIV-1 was probably introduced into the country via several routes, and thus the pattern of the HIV-1 epidemic might evolve in the near future. PMID- 11429128 TI - Variable sequences in the long terminal repeat and Its downstream region of some of HIV Type 1 CRF01_AE recently distributing among Thai carriers. AB - Human immunodeficiency virus type 1 (HIV-1) proviral DNA sequences in and downstream of the 5' long terminal repeat (LTR) were compared among samples obtained from 13 HIV-1 CRF01_AE-infected individuals in Thailand from 1998 to 1999. Eleven individuals had highly conserved sequences compared with previously reported CRF01_AE viruses. However, T cell-specific factor (TCF)-1alpha motif, which is located just beside the 3' terminus of the nef sequence, was duplicated in 2 out of the 13 subjects, one of whom had also lost the 24 nucleotides next to the 3' of the primer-binding site. Thus, several characteristics of CRF01_AE LTR and gag-leader sequence were identified in some samples recently obtained in Thailand. PMID- 11429129 TI - Full-length sequences of two CRF01_ae (subtype e) HIV type 1 isolates from 1995 samples of patients with sexually transmitted diseases in Thailand. AB - We isolated two CRF01_AE human immunodeficiency virus type 1 (95TNIH022 and 95TNIH047) from the 1995 blood samples derived from asymptomatic carriers in Ubonratchatani province of northeastern Thailand. Both isolates can replicate in peripheral blood mononuclear cells, but not in several T cell lines examined. The full-length sequences recovered from proviruses in infected cells by long-range polymerase chain reaction were determined. Phylogenetic analyses of these sequences at individual genes showed them to be closely related to those of reported CRF01_AE HIV-1, such as 1990 isolate CM240 and 1993 isolate 93TH253. Two isolates in this study also showed a similar pattern of CRF01_AE mosaicism and a similar structure at the long terminal repeat, i.e., a copy number of NF-kappaB binding sites, sequence at the TATA box, and the putative secondary structure of stem-loop in the transactivation response region. Our results showed that 1995 Thai E isolates could contribute to our understanding of the epidemiology, pathogenesis, and diagnostics of HIV-1 CRF01_AE and further to vaccine development. PMID- 11429130 TI - Rock-scissors-paper and the survival of the weakest. AB - In the children's game of rock-scissors-paper, players each choose one of three strategies. A rock beats a pair of scissors, scissors beat a sheet of paper and paper beats a rock, so the strategies form a competitive cycle. Although cycles in competitive ability appear to be reasonably rare among terrestrial plants, they are common among marine sessile organisms and have been reported in other contexts. Here we consider a system with three species in a competitive loop and show that this simple ecology exhibits two counter-intuitive phenomena. First, the species that is least competitive is expected to have the largest population and, where there are oscillations in a finite population, to be the least likely to die out. As a consequence an apparent weakening of a species leads to an increase in its population. Second, evolution favours the most competitive individuals within a species, which leads to a decline in its population. This is analogous to the tragedy of the commons, but here, rather than leading to a collapse, the 'tragedy' acts to maintain diversity. PMID- 11429131 TI - Exploitation of pollen by mycorrhizal mycelial systems with special reference to nutrient recycling in boreal forests. AB - Very large quantities of pollen are released annually by wind-pollinated trees, which dominate northern forest ecosystems. Since pollen is enriched in both nitrogen and phosphorus, this recurrent pulse of deposition constitutes a significant potential source of these elements in what are known to be severely nutrient-limited systems. Here, we demonstrate for the first time, to our knowledge, that an ectomycorrhizal fungus, Paxillus involutus, is able to scavenge effectively for nitrogen and phosphorus in pollen and to return a significant proportion of each nutrient to its autotrophic host, Betula pendula. More than 75 and 96%, respectively, of the nitrogen and phosphorus were removed from pollen in microcosms containing the mycorrhizal fungus, 29 and 25%, respectively, being transferred to the plants. In contrast, in microcosms without the mycorrhizal fungus only 42 and 35%, respectively, of nitrogen and phosphorus were lost from the pollen, presumably as a result of export by saprotrophs, and only 12 and 7%, respectively, were transferred to the plants. We hypothesize that this process of resource recapture, by contributing significantly to the ability of the trees to sustain the necessary annual investment in pollen production, will have a major impact upon their reproductive capabilities and hence 'fitness'. PMID- 11429132 TI - Testosterone influences basal metabolic rate in male house sparrows: a new cost of dominance signalling? AB - Sexually selected signals of individual dominance have profound effects on access to resources, mate choice and gene flow. However, why such signals should honestly reflect individual quality is poorly understood. Many such signals are known to develop under the influence of testosterone. We conducted an experiment in male house sparrows in which testosterone was manipulated independently during two periods: before the onset of the breeding season and prior to the autumn moult. We then measured the effects of these manipulations on basal metabolic rate and on the size of the chest bib, a sexually selected signal. The results demonstrate that testosterone simultaneously affects both signal development and basal metabolic rate in the house sparrow (Passer domesticus). This evidence, therefore, supports a novel conclusion: that testosterone-dependent signals act as honest indicators of male quality possibly because only high-quality individuals can sustain the energetic costs associated with signal development. PMID- 11429133 TI - Convergence and divergence in the evolution of aquatic birds. AB - Aquatic birds exceed other terrestrial vertebrates in the diversity of their adaptations to aquatic niches. For many species this has created difficulty in understanding their evolutionary origin and, in particular, for the flamingos, hamerkop, shoebill and pelecaniforms. Here, new evidence from nuclear and mitochondrial DNA sequences and DNA-DNA hybridization data indicates extensive morphological convergence and divergence in aquatic birds. Among the unexpected findings is a grouping of flamingos and grebes, species which otherwise show no resemblance. These results suggest that the traditional characters used to unite certain aquatic groups, such as totipalmate feet, foot-propelled diving and long legs, evolved more than once and that organismal change in aquatic birds has proceeded at a faster pace than previously recognized. PMID- 11429134 TI - From bipedalism to bicyclism: evolution in energetics and biomechanics of historic bicycles. AB - We measured the metabolic cost (C) and mechanical work of riding historic bicycles at different speeds: these bicycles included the Hobby Horse (1820s), the Boneshaker (1860s), the High Wheeler (1870s), the Rover (1880s), the Safety (1890s) and a modern bicycle (1980s) as a mean of comparison. The rolling resistance and air resistance of each vehicle were assessed. The mechanical internal work (W(INT)) was measured from three-dimensional motion analysis of the Hobby Horse and modern bicycle moving on a treadmill at different speeds. The equation obtained from the modern bicycle data was applied to the other vehicles. We found the following results. (i) Apart from the Rover, which was introduced for safety reasons, every newly invented bicycle improved metabolic economy. (ii) The rolling resistance decreased with subsequent designs while the frontal area and, hence, aerodynamic drag was fairly constant (except for the High Wheeler). (iii) The saddle-assisted body weight relief (which was inaugurated by the Hobby Horse) was responsible for most of the reduction in metabolic cost compared with walking or running. Further reductions in C were due to decreases in stride/pedalling frequency and, hence, W(INT) at the same speeds. (iv) The introduction of gear ratios allowed the use of pedalling frequencies that optimize the power/contraction velocity properties of the propulsive muscles. As a consequence, net mechanical efficiency (the ratio between the total mechanical work and C) was almost constant (0.273 +/- 0.015s.d.) for all bicycle designs, despite the increase in cruising speed. In the period from 1820 to 1890, improved design of bicycles increased the metabolically equivalent speed by threefold compared with walking at an average pace of ca. + 0.5 ms(-1) per decade [corrected]. The speed gain was the result of concurrent technological advancements in wheeled, human-powered vehicles and of 'smart' adaptation of the same actuator (the muscle) to different operational conditions. PMID- 11429135 TI - Parasite infection rather than tactile stimulation is the proximate cause of cleaning behaviour in reef fish. AB - Cleaning behaviour is a popular example of non-kin cooperation. However, quantitative support for this is generally sparse and the alternative, that cleaners are parasitic, has also been proposed. Although the behaviour involves some of the most complex and highly developed interspecific communication signals known, the proximate causal factors for why clients seek cleaners are controversial. However, this information is essential to understanding the evolution of cleaning. I tested whether clients seek cleaners in response to parasite infection or whether clients seek cleaners for tactile stimulation regardless of parasite load. Parasite loads on client fish were manipulated and clients exposed to cleaner fish and control fish behind glass. I found that parasitized client fish spent more time than unparasitized fish next to a cleaner fish. In addition, parasitized clients spent more time next to cleaners than next to control fish, whereas unparasitized fish were not attracted to cleaners. This study shows, I believe for the first time, which is somewhat surprising, that parasite infection alone causes clients to seek cleaning by cleaners and provides insight into how this behaviour evolved. PMID- 11429136 TI - Status change during adulthood: life-history by-product or kin selection based on reproductive value? AB - When dominance status predicts fitness, most adaptive models of dominance relationships among cercopithecine primate females predict lifetime maintenance of status. These models and alternative ones positing rank decline as a non adaptive by-product have remained largely untested, however, because lifetime status of older adults has been virtually unknown for natural populations. In a 25-year study of adult female savannah baboons (Papio cynocephalus), in each of three social groups, rank losses were common among the 66 females that lived past median adult age. These losses were not accounted for by loss in relative rank from group growth or by loss in absolute rank from reversals in rank between members of different maternal families or between sisters. Rather, females that had mature daughters experienced loss of dominance status to these offspring, a characteristic of all but the top-ranking matriline of each group. Among proposed hypotheses for rank reversals between adults, that of kin selection based on relative reproductive value is most clearly supported by these data. In contrast, observed patterns of rank loss are not consistent with alternative models that postulate that changes during adult lifespan are a product of accumulated risk, physical decline during ageing, or coalitionary support among females within or between matrilines. PMID- 11429137 TI - Auditory sensitivity of Hawaiian moths (Lepidoptera: Noctuidae) and selective predation by the Hawaiian hoary bat (Chiroptera: Lasiurus cinereus semotus). AB - The islands of Hawai'i offer a unique opportunity for studying the auditory ecology of moths and bats since this habitat has a single species of bat, the Hawaiian hoary bat (Lasiurus cinereus semotus), which exerts the entire predatory selection pressure on the ears of sympatric moths. I compared the moth wings discarded by foraging bats with the number of surviving moths on the island of Kaua'i and concluded that the endemic noctuid Haliophyle euclidias is more heavily preyed upon than similar-sized endemic (e.g. Agrotis diplosticta) and adventive (Agrotis ipsilon and Pseudaletia unipuncta) species. Electrophysiological examinations indicated that, compared with species less preyed upon, H. euclidias has lower auditory sensitivities to the bat's social and echolocation calls, which will result in shorter detection distances of the bat. The poor ears of H. euclidias suggest that this moth coevolved with the bat using non-auditory defences that resulted in auditory degeneration. This moth now suffers higher predation because it is drawn away from its normal habitat by the man-made lights that are exploited by the bat. PMID- 11429138 TI - Acoustic divergence in two cryptic Hipposideros species: a role for social selection? AB - We present evidence that a relatively widespread and common bat from South East Asia comprises two morphologically cryptic but acoustically divergent species. A population of the bicoloured leaf-nosed bat (Hipposideros bicolor) from Peninsular Malaysia exhibits a bimodal distribution of echolocation call frequencies, with peaks in the frequency of maximum energy at ca. 131 and 142 kHz. The two phonic types are genetically distinct, with a cytochrome b sequence divergence of just under 7%. We consider the mechanisms by which acoustic divergence in these species might arise. Differences in call frequency are not likely to effect resource partitioning by detectable prey size or functional range. However, ecological segregation may be achieved by differences in microhabitat use; the 131kHz H. bicolor is characterized by significantly longer forearms, lower wing loading, a lower aspect ratio and a more rounded wingtip, features that are associated with greater manoeuvrability in flight that may enable it to forage in more cluttered environments relative to the 142 kHz phonic type. We suggest that acoustic divergence in these species is a consequence of social selection for a clear communication channel, which is mediated by the close link between the acoustic signal and receptor systems imposed by the highly specialized nature of the hipposiderid and rhinolophid echolocation system. PMID- 11429139 TI - Immigration and the ephemerality of a natural population bottleneck: evidence from molecular markers. AB - Population bottlenecks are often invoked to explain low levels of genetic variation in natural populations, yet few studies have documented the direct genetic consequences of known bottlenecks in the wild. Empirical studies of natural population bottlenecks are therefore needed, because key assumptions of theoretical and laboratory studies of bottlenecks may not hold in the wild. Here we present microsatellite data from a severe bottleneck (95% mortality) in an insular population of song sparrows (Melospiza melodia). The major findings of our study are as follows: (i) The bottleneck reduced heterozygosity and allelic diversity nearly to neutral expectations, despite non-random survival of birds with respect to inbreeding and wing length. (ii) All measures of genetic diversity regained pre-bottleneck levels within two to three years of the crash. This rapid recovery was due to low levels of immigration. (iii) The rapid recovery occurred despite a coincident, strong increase in average inbreeding. These results show that immigration at levels that are hard to measure in most field studies can lead to qualitatively very different genetic outcomes from those expected from mutations only. We suggest that future theoretical and empirical work on bottlenecks and metapopulations should address the impact of immigration. PMID- 11429140 TI - Anomalous mole-fraction effects in recombinant and native cyclic nucleotide-gated channels in rat olfactory receptor neurons. AB - Anomalous mole-fraction effects (AMFE) were studied, using the inside-out configuration of the patchclamp technique, in both recombinant wild-type alpha homomeric rat olfactory adenosine 3',5'-cyclic monophosphate (cAMP)-gated channels (rOCNC1) expressed in human embryonic kidney cells (HEK 293) and native cyclic nucleotide-gated (CNG) channels in acutely isolated rat olfactory receptor neurons. Single-channel and macroscopic currents were activated by 200 microM and 500 microM cAMP, respectively. Macroscopic currents, measured with mixtures of Na(+)-NH(4)(+) or Cs(+)-Li(+) in the cytoplasmic bathing solution, displayed AMFE in the rOCNC1 channels at both positive and negative membrane potentials. The rOCNC1 single-channel conductance showed a distinct minimum (or maximum) in an 80% Na(+)-20% NH(4)(+) mixture (or a 60% Cs(+)-40% Li(+) mixture), but only at positive membrane potentials. Macroscopic measurements in native olfactory CNG channels with mixtures of Na(+)-NH(4)(+) indicated similar AMFE. These results suggest that both native CNG channels and recombinant alpha-homomeric channels allow several ions to be present simultaneously within the channel pore. They also further validate the dominant role of the alpha-subunit in permeation through these channels, provide the first evidence to suggest that rOCNC1 channels have multi-ion properties and further justify the use of the rOCNC1 channel as an effective model for structure-function studies of ion permeation and selectivity in olfactory CNG channels. PMID- 11429141 TI - The avian tail reduces body parasite drag by controlling flow separation and vortex shedding. AB - The aerodynamic effect of the furled avian tail on the parasite drag of a bird's body was investigated on mounted, frozen European starling Sturnus vulgaris in a wind tunnel at flight speeds between 6 and 14 m s(-1). Removal of tail rectrices and dorsal and ventral covert feathers at the base of the tail increased the total parasite drag of the body and tail by between 25 and 55%. Flow visualization and measurements of dynamic pressure in the tail boundary layer showed that in the intact bird a separation bubble forms on the ventral side of the body, and reattaches to the ventral side of the tail. This bubble is a consequence of the morphology of the body, with a rapid contraction posterior to the pelvis and hind legs. The tail and the covert feathers at its base act as a combined splitter plate and wedge to control vortex shedding and body wake development, and thereby are important to minimize drag. This hitherto unsuspected mechanism is central to understanding the morphology of the avian body, and may have had a significant influence on the evolution of avian tail morphology by pre-adapting the tail for radiation and specialization as an aerodynamic lifting structure and as an organ of communication in sexual selection. PMID- 11429142 TI - Effects of Schistocephalus solidus infection on brain monoaminergic activity in female three-spined sticklebacks Gasterosteus aculeatus. AB - The three-spined stickleback Gasterosteus aculeatus is an intermediate host of the tapeworm Schistocephalus solidus. Changes in predator avoidance, foraging and shoaling behaviour have been reported in sticklebacks infested with S. solidus, but the mechanisms underlying parasite-induced behavioural changes are not understood. Monoamine neurotransmitters are involved in the control of behaviour and central monoaminergic systems are sensitive to various stressors. Thus, the behavioural effects of S. solidus infestation might be a reflection of changes in brain monoaminergic activity in the stickleback host. The concentrations of 5 hydroxytryptamine (5-HT), dopamine (DA), norepinephrine (NE) and their metabolites 5-hydroxy-indoleacetic acid (5-HIAA), homovanilic acid (HVA) and 3 methoxy-4-hydroxyphenylglycol (MHPG) were measured in the telencephalons, hypothalami and brainstems of parasitized and non-parasitized female sticklebacks held in the laboratory. The ratios of 5-HIAA:5-HT were significantly elevated in both the hypothalami and brainstems of infected sticklebacks. The concentrations of 5-HT and NE were significantly reduced in the telencephalons of infected fish as compared with controls, but there was no elevation of metabolite concentrations. The results are consistent with chronic stress in infected fish, but may also reflect other alterations of neuroendocrine status resulting from parasite infection. PMID- 11429143 TI - The unique features of starch metabolism in red algae. AB - Red algae (Rhodophyceae) are photosynthetic eukaryotes that accumulate starch granules outside of their plastids. The starch granules from red algae (floridean starch) show structural similarities with higher plant starch granules but lack amylose. Recent studies have indicated that the extra-plastidic starch synthesis in red algae proceeds via a UDP glucose-selective alpha-glucan synthase, in analogy with the cytosolic pathway of glycogen synthesis in other eukaryotes. On the other hand, plastidic starch synthesis in green cells occurs selectively via ADP glucose in analogy with the pathway of glycogen synthesis in prokaryotes from which plastids have evolved. Given the emerging consensus of a monophyletic origin of plastids, it would appear that the capacity for starch synthesis selectively evolved from the alpha-glucan synthesizing machinery of the host ancestor and its endosymbiont in red algae and green algae, respectively. This implies the evolution of fundamentally different functional relationships between the different subcellular compartments with regard to photosynthetic carbon metabolism in these organisms. It is suggested that the biochemical and molecular elucidation of floridean starch synthesis may offer new insights into the metabolic strategies of photosynthetic eukaryotes. PMID- 11429144 TI - Recombination confounds interpretations of Wolbachia evolution. AB - Wolbachia are vertically transmitted bacteria known from arthropods and nematode worms, which are maintained in host populations because they either physiologically benefit infected individuals or parasitically manipulate their reproduction. The different manipulation phenotypes are scattered across the Wolbachia phylogeny, suggesting that there have been multiple evolutions of similar phenotypes. This conclusion relies on the assumption of an absence of recombination between bacterial strains, so that the gene used to reconstruct the phylogeny reflects the evolutionary history of the genes involved in the trait. We tested for recombination by reconstructing the phylogeny of two Wolbachia genes from seven B-subdivision strains. The two genes produced mutually incompatible topologies, indicating that these lineages are subject to genetic recombination. This means that many evolutionary patterns inferred from Wolbachia phylogenies must be re-evaluated. Furthermore, recombination may be an important feature both in the evolution of the manipulation phenotypes and avoidance of Muller's ratchet. Finally, we discuss the implications of recombination for attempts to genetically engineer Wolbachia for use in the control of crop pests and human pathogens. PMID- 11429145 TI - A polymorphic effect of sexually differential production costs when one parent controls the sex ratio. AB - R. A. Fisher's sex ratio theory predicts that if sons and daughters cost fixed amounts of resources to raise and parents have fixed amounts to invest, then the numerical sex ratio of a panmictic population will evolve to be inversely proportional to relative cost. However, the theory assumes control by both parents. We show that allowing one parent to control the sex ratio biases it further from parity than Fisher's theory predicts. Quantitatively, the additional bias towards the cheaper sex depends only very weakly on which sex is in control. Qualitatively, however, the effect is very strong: a monomorphic, mixed-brood strategy evolves only if the more expensive sex is in control. If the controlling sex is cheaper to raise, then the sex ratio is instead achieved through a polymorphism of single-sex broods. Such polymorphisms are seldom observed in nature, generating the prediction that wherever the sexes are not equally costly, sex ratio is usually either under biparental control or under uniparental control by the more expensive sex. However, such polymorphisms do occur, and some of them may be explained by our model. PMID- 11429146 TI - Schwann cell response to micropatterned laminin surfaces. AB - In the peripheral nervous system, Schwann cells are closely associated with, and play key roles in, the development, maintenance, and regeneration of peripheral neurons. Following injury, Schwann cell orientation may also play a role in guiding regenerating axons. To aid in the investigation of these interactions between Schwann cells and growing neurites, we have developed a method of controlling Schwann cell placement and orientation in vitro by using microlithographically patterned laminin substrates, alternating 20-microm regions of laminin with bovine serum albumin (BSA) stripes. The Schwann cells predominantly attached and elongated on the laminin stripes and organized into multicellular aggregates that were oriented with the micropattern. A detailed analysis of Schwann cell aggregate orientation and shape demonstrated a strong dependence on time. At 1 h after seeding the cells, 70% of the aggregates were oriented with respect to the micropattern; 94% were oriented at 24 h. Variations in laminin concentration and seeding density were also investigated. The only significant differences in Schwann cell response occurred 1 h after seeding (the earliest time point the cultures were observed), and the main factor controlling the cellular orientation appeared to be the presence of the laminin-BSA interface. This ability to control cell orientation and placement provides a tool for future investigations of Schwann cell-neuronal interactions in vitro. PMID- 11429147 TI - Development of a three-dimensional transmigration assay for testing cell--polymer interactions for tissue engineering applications. AB - The ability of synthetic or natural scaffolds to support invasion of cells from surrounding tissue is a key parameter for tissue engineering (TE). In this study, the migration of fibroblasts, chondrocytes, and osteoblasts into biodegradable polymer scaffolds was evaluated using a novel, three-dimensional (3-D) transmigration assay. This assay is based on a cell-populated contracted collagen lattice with a biodegradable polymer scaffold implanted at the center of the collagen gel. Cell migration into the scaffolds was assessed both quantitatively and qualitatively following various time lengths in culture using image analysis. Chondrocytes, incorporated within the collagen lattice, migrated into polymer scaffolds, when cultured both statically or in a rotating bioreactor. However, the bioreactor cultures resulted in a significantly greater cell invasion as compared to static cultures. There was a cell density-dependent osteoblast migration from collagen lattice into polymer scaffold, when tested in the transmigration assay. In addition, polymer scaffolds, treated with or without recombinant human platelet-derived growth factor (rh-PDGF-BB) were evaluated for fibroblast migration. The presence of rh-PDGF-BB resulted in significantly greater fibroblast invasion as compared to untreated scaffolds. Our studies suggest that the transmigration model provides a rapid system for testing cell invasion of potential scaffolds for tissue engineering applications. PMID- 11429148 TI - Analysis of integrin expression in U2OS cells cultured on various calcium phosphate ceramic substrates. AB - Earlier we observed that calcium phosphate (Ca-P)-coated implant substrates stimulated the differentiation of osteoblast-like cells compared to uncoated substrates. This suggests that this difference in osteogenic induction is due to the chemical composition of the substratum. We hypothesized that Ca-P coatings modulate integrin expression patterns, because those receptors are the sensors of the cell. Therefore, in the present study we quantitatively analyzed integrin expression of osteosarcoma cells and their proliferation behavior on various well defined Ca-P substrates. For this study we used the osteosarcoma cell line U2OS. Five groups of substrates were used: thermanox (Th), uncoated titanium (Ti), dense sintered hydroxyapatite (HA), and two Ca-P-coated titanium discs (TiHA-O% and TiHA-5%). At day 5, cell numbers were significantly lower (p < 0.05) for both types of Ca-P-coated titanium substrates compared to the other substrates. There were no significant differences between HA and uncoated titanium. From day 5 to 8, accumulated cell number was ranking highest to lowest HA > Th = Ti > TiHA-0% > TiHA-5%. Integrin expression at day 5 and day 8 of incubation was analyzed by flow cytometry for integrin subunits beta 1, alpha 3, alpha 4, alpha 5, alpha 6, and alpha v. Fluorescence-activated cell sorting (FACS) analysis showed that the cells express high levels of beta 1, low levels of alpha 4, alpha 5, and alpha 6, and moderate levels of alpha 3 and alpha v integrin subunits on the various biomaterial substrates. Minor differences in integrin expression between the various substrates were seen. Therefore, the observed differences in proliferation between the coatings may reside in modulating the functional properties of integrins. PMID- 11429149 TI - Mechanisms of enhanced osteoblast adhesion on nanophase alumina involve vitronectin. AB - The role, including concentration, conformation, and bioactivity, of adsorbed vitronectin in enhancing osteoblast adhesion on nanophase alumina was investigated in the present study. Vitronectin adsorbed in a competitive environment in the highest concentration on nanophase alumina compared to conventional alumina. Enhanced adsorption of vitronectin on nanophase alumina was possibly due to decreased adsorption of apolipoprotein A-I and/or increased adsorption of calcium on nanophase alumina. In a novel manner, the present study utilized surface-enhanced Raman scattering (SERS) to determine the conformation of vitronectin adsorbed on nanophase alumina. These results provided the first evidence of increased unfolding of vitronectin adsorbed on nanophase alumina. Increased adsorption of calcium on nanophase alumina may affect the conformation of adsorbed vitronectin specifically to promote unfolding of the macromolecule to expose cell-adhesive epitopes recognized by specific cell-membrane receptors. Results of the present study also provided evidence of dose-dependent inhibition of osteoblast adhesion on nanophase alumina pretreated with vitronectin following preincubation (and thus blocking respective cell-membrane receptors) with either Arginine-Glycine-Aspartic Acid-Serine (RGDS) or Lysine-Arginine-Serine-Arginine (KRSR). These events, namely, enhanced vitronectin adsorption, comformation, and bioactivity, may explain the increased osteoblast adhesion on nanophase alumina. PMID- 11429150 TI - Muristerone A-induced nerve growth factor release from genetically engineered human dermal fibroblasts for peripheral nerve tissue engineering. AB - In this study, human dermal fibroblasts (hDFBs) were genetically modified to release human nerve growth factor (NGF) using an ecdysone-inducible system. NGF cDNA was inserted into the pIND vector and then hDFBs were cotransfected with pIND-NGF and pVgRXR. Muristerone A, an analog of ecdysone, was used as the inducing agent. NGF release from transfected hDFBs was assessed in vitro and in vivo. Transfected hDFBs in the presence of Muristerone A possessed a maximal in vitro release of 8.5 +/- 0.4 pg of NGF/mL per 10(3) cells, demonstrating significantly higher NGF levels compared to control hDFBs. The in vitro release rate curve for transfected hDFBs in the presence of Muristerone A exhibited a maximum of 5.1 +/- 0.2 ng NGF/10(6) cells/day. A PC-12 bioassay demonstrated that the in vitro NGF released is bioactive. When transfected hDFBs in the presence of Muristerone A were placed in vivo in nude rats, NGF levels reach 2074 +/- 257 pg/mL and 1620 +/- 132 pg/mL at 24 and 48 h, respectively. These levels were significantly higher than negative control and wound fluid levels. Results support further in vivo investigation of this molecular "on" switch for peripheral nerve regeneration. PMID- 11429151 TI - Validation of a simple histological-histochemical cartilage scoring system. AB - In this study, we assessed the validity of a subjective histological histochemical scoring system as compared to an automated histomorphometry program for analyzing cartilage repair tissue. In the first part of the study, we assessed the ability of the human eye to estimate the percent cartilage in a histological section. Twenty-nine rabbit periosteal explants that had been cultured in agarose transforming growth factor-beta (TGF-beta) were selected so that the percentage of cartilage in the specimens was distributed equally from 0% to 100%. Color photomicrographs were evaluated by 5 expert observers who gave a visual estimate of the percent cartilage. There was a strong correlation between the estimated and actual percent cartilage (R(2) = 0.92, p < 0.0001) and among the observers (I.C.C. = 0.89). On average, the estimated percent cartilage was within ten percent of the actual percent measured. In the second part, we compared the data derived using a simple cartilage score with those obtained by automated image analysis. The histological slides from 159 explants cultured under various experimental conditions (14 treatment groups) in two different experiments were analyzed. The cartilage content was estimated visually and a score from 0 to 3 was assigned. A previously validated, computerized image analysis system was used to measure the actual percent cartilage. Statistical analyses revealed a good linear regression (R(2) = 0.84, p = 0.0001), and even better polynomial correlation between the actual measurement and the score (R(2) = 0.88, p = 0.0001). These data demonstrate the validity of a simple histological histochemical subjective scoring system. A computerized automated program such as the one employed in this study is preferable due to its many advantages. However, a subjective scoring system may be appropriate to use when the funding and expertise required for a computerized image analysis program are not available. PMID- 11429152 TI - Kinetic study of the replacement of porcine small intestinal submucosa grafts and the regeneration of meniscal-like tissue in large avascular meniscal defects in dogs. AB - Porcine small intestinal submucosa (SIS) was used to replace large, avascular defects in the medial menisci of dogs. Twelve dogs received SIS grafts and 3 dogs were left untreated as controls. Dogs were evaluated at 4, 8, and 12 weeks by means of lameness scoring and ultrasonography. Dogs were sacrificed at 1, 6, or 12 weeks after implantation, and the tissue at the site of meniscal resection was evaluated for gross and histologic appearance, cross-sectional and surface area, and collagen types I and II. The femoral and tibial condyles were assessed for articular cartilage damage. Control dogs were significantly more lame than grafted dogs 8 and 12 weeks after instrumentation. Grafted dogs' replacement tissue appeared meniscal-like when evaluated grossly and ultrasonographically 12 weeks after instrumentation. The amount of replacement tissue was significantly greater in both cross-sectional and surface area for grafted dogs than for controls at all time points. Histologically, the SIS biomaterial could be identified in all grafted dogs at 1 week post-implantation, but in none at 6 weeks post-implantation. Subjectively, grafted dogs' replacement tissue was histologically superior to that of controls with respect to tissue type, organization, and architecture. Collagen types I and II immunoreactivity in grafted menisci were similar to that of normal menisci. Control dogs had significantly more articular cartilage damage than grafted dogs. SIS appears to induce regeneration of meniscal-like tissue in large, avascular meniscal defects in dogs, resulting in superior clinical function and articular cartilage protection compared to ungrafted controls. PMID- 11429153 TI - Establishment of heterotropic liver tissue mass with direct link to the host liver following implantation of hepatocytes transfected with vascular endothelial growth factor gene in mice. AB - One of the major goals of tissue engineering is to establish an integrated organ in vivo. We have previously shown that transfection of vascular endothelial growth factor (VEGF) gene into hepatocytes promotes tissue formation by engrafted cells. Here we show that tissue growth was significantly enhanced by co transplantation of hepatocyte growth factor (HGF) and tumor necrosis factor-alpha (TNF alpha) gene transfected hepatocytes with VEGF-gene transfected cells, but tissue islands were scattered nonspecifically in the abdomen of mice. The result brought us forward to the next step to establish an integrated mass and structural formation of liver tissue. We entrapped VEGF gene transfected hepatocytes in a nylon mesh bag and intraperitoneally engrafted close to the liver. Three weeks later, the bag was covered by a thick network of blood vessels, compared to the control. Histological examination showed that the blood vessels penetrated the parenchyma of the engrafted bag and formed a well developed vessel network in the region. The use of hepatocytes from lacZ transgenic mice and PCR analysis demonstrated survival and albumin production by hepatocytes in the engrafted bag. Our model can potentially be developed into a heterotropic artificial liver with direct access to the host blood circulation. PMID- 11429154 TI - Liver tissue engineering: a role for co-culture systems in modifying hepatocyte function and viability. AB - A major limitation in the construction of a functional engineered liver is the short-term survival and rapid de-differentiation of hepatocytes in culture. Heterotypic cell-cell interactions may have a role to play in modulating long term hepatocyte behavior in engineered tissues. We describe the potential of 3T3 fibroblast cells in a co-culture system to modulate function and viability of primary isolated rat hepatocytes. Over an 18-day period after isolation, hepatocytes in pure culture rapidly declined in viability, displayed sparse bile canaliculi, and lost two function markers, the secretion of albumin and ethoxyresorufin O-dealkylase (EROD) activity. In comparison, the hepatocytes within the co-cultures maintained viability, possessed well-formed canalicular systems, and displayed both functional markers. Fixed 3T3 cells or 3T3 cell conditioned medium did not substitute for the viable 3T3 cell co-culture system in preserving hepatocyte viability and functionality. PMID- 11429155 TI - The use of fibrin beads for tissue engineering and subsequential transplantation. AB - New biological technologies such as tissue engineering procedures require the transplantation of functionally active cells within supportive carrier matrices. This paper describes a sequential culture procedure for different types of cells. The technique includes the initial preparation of a mixed alginate-fibrin vehicle that guaranteed an initial cell proliferation and differentiation to establish a stable matrix structure, and the subsequent removal of the alginate component prior to transplantation to circumvent the problem of missing bioresorbability. The resulting biodegradable carrier is mechanically stable and promotes further tissue maturation. Chondrocytes, periosteal-derived cells, as well as nucleus pulposus cells were entrapped in fibrin-alginate beads and in fibrin beads. The results indicate a promising technical approach to create stable transplants for reconstructive surgery of cartilage and bone. PMID- 11429156 TI - In vitro modulation of adhesion molecules, adhesion phenomena, and fluid phase endocytosis on human umbilical vein endothelial cells and brain-derived microvascular endothelium by IFN-beta 1a. AB - Administration of interferon-beta (IFN-beta) in multiple sclerosis (MS) patients provides clinical benefits, although its mechanism(s) of action are not completely understood. We addressed the issue of whether concentrations of IFN beta1a close to those reached in the serum of treated MS patients could modulate either adhesion molecules or adhesion of peripheral blood mononuclear cells (PBMC) as well as fluid phase endocytosis (FPE) in human umbilical vein endothelial cells (HUVEC) and in brain-derived microvascular endothelial cells (HBMEC). Adhesion was assessed by flow cytometry, and FPE was evaluated by peroxidase uptake. In our study, 200 U/ml IFN-beta1a induced a reduction in adhesion of PBMC to HUVEC. The information reported herein may contribute to further elucidating some of the mechanisms of action of IFN-beta on vascular endothelium. PMID- 11429157 TI - Neuropsychiatric effects and type of IFN-alpha in chronic hepatitis C. AB - Chronic hepatitis is often associated with neuropsychiatric disorders. Interferon (IFN) is the drug most widely used to treat this disease, and its side effects, such as depression, often involve the central nervous system (CNS). Symptoms include a slowing down of psychomotor functions, loss of interest, frontal lobe dysfunction, parkinsonism, and delirium. The occurrence of these complications calls for dropping out of IFN treatment or for a significant dose reduction and administration of antidepressants. Efficacy and side effects vary on the basis of the IFN type employed. The aim of our study was to evaluate if the frequency, form, and degree of depression induced are related to the type of IFN employed. We studied 96 patients with chronic hepatitis C. Our study series was divided into four groups according to the type of IFN-alpha administered. Depression degree was clinically evaluated using the Hamilton Depression Rating Scale (HAM D). All patients were tested before treatment and 1, 3, and 6 months (15 days after the end of treatment) later. Our results showed that the type of IFN used seemed to influence the depression onset rate, with the leukocyte type inducing the lowest level of depression. However, when a number of symptoms associated with the depression were considered, the results of other types of IFN-alpha were found to be better. Use of the most suitable type of IFN-alpha could thus lead to more personalized treatment, with fewer side effects. The type of IFN used seems to influence the psychological side effects and the adaptation rate to therapy. It would be appropriate to choose the type of IFN on the basis of a neuropsychiatric assessment carried out before treatment. PMID- 11429158 TI - Participation of type I interferon in the decreased virulence of the UL13 gene deleted mutant of herpes simplex virus type 1. AB - We isolated a UL13 gene-deleted mutant of the herpes simplex virus type 1 (HSV-1) strain VR3 (VRDelta13) and its revertant virus (VRDelta13R). This deletion mutant still had virus host shutoff (vhs) activity, although a previous report had suggested the possibility of a functional relation between the UL13 product, that is protein kinase (PK), and vhs activity. We compared the in vivo growth of these viruses in BALB/c mice. VRDelta13 was cleared in the early period of intraperitoneal infection. VRDelta13 had a higher sensitivity to the mouse type I interferon (IFN) and showed a higher level of IFN induction in the study period of infection than did VR3 and VRDelta13R. These results suggest that a nonspecific antiviral response (i.e., the IFN system) may contribute to this rapid inhibition of viral replication in vivo. PMID- 11429159 TI - Cellular responses in chickens treated with IFN-alpha orally or inoculated with recombinant Marek's disease virus expressing IFN-alpha. AB - Mammalian type I interferons (IFN-alpha/beta) are potent mediators of innate antiviral immune responses, in particular through enhancement of natural killer (NK) cell cytotoxicity. Recently, chicken IFN-alpha (ChIFN-alpha) has been identified and shown to ameliorate Newcastle disease virus (NDV) infection when given to chickens at relatively high concentrations in the drinking water. In this report, the effect of recombinant ChIFN-alpha (rChIFN-alpha) on NK cell cytotoxicity was examined using (51)Cr-release assays. NK cell cytotoxic activity was also analyzed following inoculation with attenuated Marek's disease virus (MDV) serotype 1 strain R2/23 and a recombinant MDV (parent strain R2/23) expressing ChIFN-alpha [rMDV(IFN-alpha)]. Treatment of chickens with high doses of rChIFN-alpha in the drinking water significantly decreased NK cell cytotoxicity compared with untreated chickens over a 7-day period. Inoculation of chickens with R2/23 significantly decreased NK cell cytotoxicity as well, whereas the rMDV(IFN-alpha) had no effect on NK cell cytotoxicity. Treatment of chicken embryo cell cultures with rChIFN-alpha inhibited replication of the very virulent MDV RB-1B strain in vitro, and oral treatment of chickens with rChIFN-alpha reduced MDV R2/23 replication in vivo. PMID- 11429160 TI - Monocyte chemoattractant protein-1 expression by osteoblasts following infection with Staphylococcus aureus or Salmonella. AB - Two common pathogens of bone, Staphylococcus aureus and Salmonella, were investigated for their ability to induce chemokine expression in bone-forming osteoblasts. Cultured mouse or human osteoblasts could rapidly respond to bacterial infection by upregulating the mRNA encoding the chemokine, monocyte chemoattractant protein-1 (MCP-1). This rapid induction occurred on infection with either the gram-positive pathogen, S. aureus, or the gram-negative pathogen, Salmonella. Increased mRNA expression translated into MCP-1 secretion by cultured mouse or human osteoblasts in response to viable bacteria, whereas UV-killed bacteria were less effective in stimulating chemokine secretion. There was a dose response relationship observed between the amount of input bacteria and increases in MCP-1 secretion. Immunohistochemical staining of infected osteoblasts indicated that the majority of cells could express MCP-1, with some osteoblasts having a higher intensity of staining than others. Organ cultures of mouse calvaria (skullcap) bone showed increases in MCP-1 immunostaining following bacterial infection. The immunoreactive MCP-1 in infected calvaria localized to areas containing active osteoblasts. Taken together, these studies demonstrate a conserved osteoblast-derived MCP-1 response to two very different pathogens of bone. PMID- 11429161 TI - Molecular cloning, characterization, and expression of porcine Fas ligand (CD95 ligand). AB - We isolated and sequenced cDNA that contained the coding sequence of porcine Fas ligand (FasL). Using mixed oligonucleotide primers based on the 5' and 3' nucleotide sequences conserved among human, murine, and rat FasL, we performed the reverse transcription polymerase chain reaction (RT-PCR) with total RNA prepared from porcine thymocytes stimulated with 5 microg/ml concanavalin A (ConA) to clone the cDNA of porcine FasL. The open reading frame (ORF) of porcine FasL cDNA was 849 base pairs (bp) in length and encoded 282 amino acids. The predicted amino acid sequence was 85.5%, 76.6%, and 75.5% homologous to the predicted human, murine, and rat FasL, respectively. The recombinant porcine FasL expressed by recombinant baculovirus containing the whole coding sequences of porcine FasL showed cytotoxic effect and induced apoptosis in porcine renal tubular cell line PK-15 cells sensitized by cycloheximide (CHX), which was confirmed by MTT assay, DNA fragmentation assay, and TUNEL staining, respectively. Furthermore, the mRNA expression of porcine FasL in porcine peripheral blood lymphocytes (PBL) was induced by porcine interleukin-18 (IL-18). These results indicate that porcine FasL identified in this study is biologically functional and has the ability to induce apoptosis as reported in other species. PMID- 11429162 TI - Differential effects of phosphatidylinositol-3/Akt-kinase inhibition on apoptotic sensitization to cytokines in LNCaP and PCc-3 prostate cancer cells. AB - Alterations in phosphatidylinositol 3'-kinase (PI3'-kinase) and Akt activation frequently occur in prostate cancer and may disrupt apoptotic induction by such cytokines as tumor necrosis factor (TNF) and TNF-related apoptosis-inducing ligand (TRAIL). To examine the role of PI3' phosphorylation in the cellular response to cytokines, two prostate cancer cell lines with constitutively activated PI3'-kinase cascades (LNCaP and PC-3) were examined for direct sensitivity to cytokines. TNF or TRAIL alone failed to activate apoptosis in either LNCaP or PC-3 cells, and drug-mediated inhibition of the PI3k/Akt cascade caused only minimal activation of apoptosis in either cell line. Suppression of PI3'-kinase/Akt signaling markedly enhanced the apoptotic activity of both TNF and TRAIL in LNCaP cells but not in PC-3 cells. Adenovirus-mediated PTEN/MMAC1 expression in LNCaP cells reduced Akt activation, activated apoptosis, and sensitized cells to TNF but not to TRAIL. Together, these results suggest that PI3'-kinase signaling inhibits both TNF-mediated and TRAIL-mediated apoptosis but may represent one of several apoptotic resistance mechanisms that inhibit cytokine-mediated killing of prostate cancer cells. PMID- 11429163 TI - CHOP, a basic leucine zipper transcriptional factor, contributes to the antiproliferative effect of IL-1 on A375 human melanoma cells through augmenting transcription of IL-6. AB - Interleukin-1 (IL-1) inhibits the proliferation of A375 human melanoma cells. We have demonstrated previously that p38 mitrogen-activated protein kinase (MAPK) mediated the antiproliferative effect of IL-1 partially through the downregulation of activity and protein level of ornithine decarboxylase (ODC). In this study, we investigated the role of CCAAT/enhancer-binding protein (C/EBP) homologous protein (CHOP), one of the p38 MAPK target transcriptional factors. The mRNA level of CHOP was not affected by IL-1 treatment in A375-6 cells. Unexpectedly, CHOP was constitutively phosphorylated, and IL-1 or p38 MAPK inhibitor, SB203580, did not affect the phosphorylation level. However, A375-6 cells exhibited enhanced sensitivity to IL-1 by transfecting CHOP expression plasmid and reduced sensitivity to IL-1 by antisense CHOP mRNA expression plasmid. Furthermore, CHOP appeared to regulate positively IL-6 production at the transcriptional level. The experiments using CHOP muteins revealed that dimerization ability - but not p38 MAPK-dependent phosphorylation or DNA binding activity - is important for the IL-6 inducing activity of CHOP. These results indicate that CHOP contributes to the IL-1 growth-inhibitory signal through augmenting IL-6 production. PMID- 11429164 TI - Recombinant, soluble LIGHT (HVEM ligand) induces increased IL-8 secretion and growth arrest in A375 melanoma cells. AB - The heterotrimeric lymphotoxin alpha(1)beta(2) (LTalpha(1)beta(2)) complex and LIGHT, a new member of the tumor necrosis factor (TNF) superfamily, have been identified as membrane-anchored ligands for the LTbeta receptor (LTbetaR), a member of the TNF receptor (TNFR) superfamily. Although some of the biologic activities of this receptor have been described using either soluble LTalpha(1)beta(2) as a ligand or agonistic monoclonal antibodies (mAb), very little is known about the signaling of LIGHT via the LTbetaR. To gain more insight into the biologic functions of LIGHT, we generated a recombinant soluble form of human LIGHT (rsHuLIGHT). We demonstrate here that this rsHuLIGHT is capable of binding to the LTbetaR. Interestingly, receptor-mediated ligand precipitation analysis revealed that rsHuLIGHT bound only to human LTbetaR but not to mouse LTbetaR, indicating a species-specific receptor ligand interaction. Activation of A375 human melanoma cells by rsHuLIGHT induced an increased secretion of interleukin-8 (IL-8). Furthermore, rsHuLIGHT caused growth arrest of A375 cells even in the absence of interferon-gamma (IFN-gamma). PMID- 11429165 TI - The differential expression of multiple isoenzyme forms during stage conversion of Toxoplasma gondii: an adaptive developmental strategy. AB - The apicomplexan parasite Toxoplasma gondii has the ability to switch between a rapidly replicating tachyzoite and a slowly dividing encysted bradyzoite within its intermediate hosts such as humans or other warm-blooded vertebrates. It is likely that in vivo, the tachyzoites differentiate into encysted bradyzoites in response to the immune system attack during disease progression. As part of a developmental strategy and, in order to survive within infected hosts, T. gondii tachyzoites undergo profound metabolic and morphological changes by differentiating into encysted bradyzoites. Bradyzoites are characterised by their resistance to both the immune system and chemotherapy. The stimulus that triggers Toxoplasma encystation and the molecular mechanisms triggering the switch from tachyzoite to bradyzoite remain unknown. It is very important to elucidate these mechanisms since bradyzoites within tissue cysts are not only the source of infection transmitted from domestic animals to humans, but can also be converted into tachyzoites that are the cause of fatal toxoplasmic encephalitis in acquired immunodeficiency syndrome patients. In this review, I focus on recent efforts towards the characterisation of genes that encode several stage-specific isoenzymes. The picture emerging from these studies is that stage-specific expression of isoenyzmes having different biochemical properties accompanies the interconversion of tachyzoite into bradyzoite, and vice versa. It can be hypothesised that the difference found between these enzymatic activities may be instrumental in maintaining some major parasitic metabolisms such as glycolysis in pace with the stage-specific requirements of carbohydrate or polysaccharide biosynthesis. PMID- 11429166 TI - Acidic ribosomal P proteins are phosphorylated in Trypanosoma cruzi. AB - Trypanosoma cruzi ribosomes from epimastigote forms were purified as determined by electron microscopy and isoelectrofocusing was used to analyse this purified ribosome fraction. Silver stained gels revealed that acidic proteins are present in at least 10 different isoforms, in accord with previous cloning studies. To detect phosphorylation, in vitro phosphorylation assays using the recombinant protein TcP2beta-mbp were carried out. The results showed that T. cruzi cytosolic fraction possesses protein kinase activity able to phosphorylate the recombinant protein. Purified ribosomes contain protein kinases that could also phosphorylate the recombinant protein TcP2beta-mbp. Labelling parasites with [(32)Pi] in a phosphate free medium demonstrated that ribosome proteins, recognised with a specific mouse antiserum against recombinant TcP2beta proteins, are phosphorylated in vivo. All these results suggest that in vivo phosphorylation of ribosome TcP2beta proteins are mediated by protein kinase(s) not yet identified. PMID- 11429167 TI - Life cycle of the tick Ixodes uriae in penguin colonies: relationships with host breeding activity. AB - A survey of the temporal pattern of population structure and feeding activity of the seabird tick Ixodes uriae was conducted for the first time in two host species colonies: King penguin (Aptenodytes patagonicus halli) and Macaroni penguin (Eudyptes chrysolophus chrysolophus). The life cycle of the tick was investigated over 3 years in a King penguin colony and 2 years in a Macaroni penguin colony at Possession Island (Crozet Archipelago). There was a marked seasonal feeding activity pattern of ticks in both host species, connected with the presence of birds during the breeding season. Although the King penguin colonies were occupied throughout the year by birds, the favourable period for engorgement was limited to 3.5-4.5 months, and almost all the ticks overwintered in the unengorged state. Consequently, I. uriae probably completed its life cycle over 3 years in King penguin colonies. In contrast, this life cycle could be shortened to 2 years in Macaroni penguin colonies, as a result of a different timetable of the presence of birds for breeding and moulting. The relationships between such plasticity and the host behaviour and subantarctic climatic conditions are discussed. PMID- 11429168 TI - Complete development and long-term maintenance of Cryptosporidium parvum human and cattle genotypes in cell culture. AB - This study describes the complete development (from sporozoites to sporulated oocysts) of Cryptosporidium parvum (human and cattle genotypes) in the HCT-8 cell line. Furthermore, for the first time the complete life cycle was perpetuated in vitro for up to 25 days by subculturing. The long-term maintenance of the developmental cycle of the parasite in vitro appeared to be due to the initiation of the auto-reinfection cycle of C. parvum. This auto-reinfection is characterised by the production and excystation of new invasive sporozoites from thin-walled oocysts, with subsequent maintenance of the complete life cycle in vitro. In addition, thin-walled oocysts of the cattle genotype were infective for ARC/Swiss mice but similar oocysts of the human genotype were not. This culture system will provide a model for propagation of the complete life cycle of C. parvum in vitro. PMID- 11429169 TI - Quantification of the crowding effect during infections with the seven Eimeria species of the domesticated fowl: its importance for experimental designs and the production of oocyst stocks. AB - The 'crowding effect' in avian coccidia, following administration of graded numbers of sporulated oocysts to naive hosts, is recognisable by two characteristics. First, increasing doses of oocysts give rise to progressively higher oocyst yields, until a level of infection is reached (the 'maximally producing dose') above which further dose increases result in progressive decreases in oocyst yields. Second, the number of oocysts produced per oocyst administered (the 'reproductive potential') tends to decrease as the oocyst dose is increased. The dose that gives the maximal reproductive potential is the 'crowding threshold' and doses exceeding this are 'crowded doses'. Graded doses of Eimeria acervulina, Eimeria brunetti, Eimeria maxima, Eimeria mitis, Eimeria necatrix, Eimeria praecox or Eimeria tenella were given to chickens of the same breed, sex and age, reared on the same diet, under identical management. The two characteristics of the crowding effect were demonstrated graphically and, by interpolation, the estimated crowding thresholds were 903, < or =16, 39, < or =14, < or =16, < or =16 or 72 sporulated oocysts, respectively, for the seven Eimeria species enumerated above. This is apparently the first report of definitive experiments to quantify a crowding effect in E. brunetti, E. maxima, E. mitis, E. necatrix and E. praecox. Maximum experimental reproductive potentials were considerably lower than the theoretical reproductive potentials for all seven species. The interaction between availability of host intestinal cells and immunity contributing to the crowding effect is discussed. Standard curves obtained under specified conditions should be used to estimate appropriate infective doses for experimental designs or in vivo production of oocyst stocks. For experiments on effects of chemotherapy or immunisation on oocyst production, an infective dose lower than the crowding threshold should be used. For efficient production of laboratory or factory oocyst stocks, the maximally producing dose (which is greater than the crowding threshold), should be used. PMID- 11429170 TI - Modelling host cell availability and the crowding effect in Eimeria infections. AB - Within-host mathematical models of Eimeria maxima and Eimeria praecox infections of the chicken are presented and used to investigate the role of host cell availability as a possible determinant of the so-called 'crowding effect'; whereby the fecundity of the parasites decreases as infectious dose increases. Assumptions about the number of available host cells, the average lifespan of these cells and the age structure within the host-cell population were made and mathematical models were constructed and combined with experimental data to test whether these conditions could reproduce the crowding effect in the two species. Experimental data demonstrated that crowding during in vivo infections was apparent following very low infectious doses, but none of the models could adequately reproduce crowding at the same doses while maintaining realistic estimates of the dynamics of the enterocyte pool. However, both the size and lifespan of the enterocyte pool were demonstrated to have substantial effects on the fecundity of the infections, particularly at higher doses. These data indicate that host cell availability cannot be solely responsible for the crowding effect. Alternative factors such as the influence of the primary immune response to the parasite may also be explored using within-host models and other applications of these models are discussed. PMID- 11429171 TI - Large sequence variation at two microsatellite loci among zoonotic (genotype C) isolates of Cryptosporidium parvum. AB - The genetic polymorphism among 57 Cryptosporidium parvum isolates belonging to genotype 'C' was studied by PCR amplification and the sequencing of two microsatellite loci (ML1 and ML2). A comparative analysis of DNA sequences showed the presence of three (ML1-238, ML1-226, and ML1-220) and seven (ML2-231, ML2 229, ML2-227, ML2-213, ML2-193, ML2-191, and ML2-187) different alleles at these two loci. Alleles differed by expansions/contractions of the microsatellite repeats that generated length polymorphisms. Some alleles were found to be associated with infections of all examined hosts (calf, kid, lamb, and human), whereas others were either associated with a single host, or were geographically restricted. When considering the information from both loci, some preferential associations between alleles are apparent. These data confirm the utility of microsatellite markers for the molecular identification of C. parvum, which is of particular relevance in the investigation of the source of infection of outbreaks and single cases, as well as for genetic studies. PMID- 11429172 TI - Avidity analysis of the human immune response to a chitin binding protein of Toxoplasma gondii. AB - A 30-33 kDa electroeluted fraction of T. gondii tachyzoites improved discrimination between acute and chronic phase sera when used instead of the whole tachyzoite extract in an avidity-ELISA. In order to identify the components of these fractions, crude tachyzoite antigen was fractionated by anionic exchange chromatography. The 30-33 kDa antigen cluster eluted in the not-bound fraction could account for a large proportion of the antibody response against the 30-33 kDa electroeluted fraction. According to the N-terminal sequence data, this antigen fraction is composed mainly of SAG1 and another protein with high homology to chitin binding proteins from plants. PMID- 11429173 TI - Stability and reproductive fitness of Schistosoma mansoni isolates with decreased sensitivity to praziquantel. AB - These studies are focused on schistosomes derived from human infections not cured by three successive doses of praziquantel that also produced infections in mice that were significantly more difficult to cure than infections with control worms. Half (three of six) of these isolates retained their decreased response to praziquantel after multiple passages through the life-cycle in the absence of therapeutic pressure. Two of the isolates, including the one initially least sensitive to praziquantel; reverted, to a sensitivity not significantly different from controls. For example, the EE6 isolate initially required 680 mg/kg praziquantel to affect a 50% reduction in worm load in murine infections, but after only six passages through the life cycle over 5 years this was reduced to 113 mg/kg, not different from control infections. The stability of some of the isolates and the reversion of others indicates that the biological or genetic factors conferring decreased praziquantel response varies among the isolates. The three isolates that retained decreased sensitivity to praziquantel all showed compromises in reproductive fitness in the laboratory, expressed most frequently as a decreased cercarial production from snails infected with those isolates compared to controls. For example, the total cercarial production of snails infected with the EE10 isolate was only 57% that of controls. The reversion of some of the isolates to a praziquantel sensitive state and the decreased reproductive fitness of those that did not revert suggest that there is some biological cost associated with the relative praziquantel insensitivity of these worms, which could help limit the impact of such isolates in the field. Infections with the less sensitive isolates also produced significantly less circulating schistosomal antigen in mice, suggesting that a decrease in the host immune response elicited by these worms could be one of the factors contributing to the diminished praziquantel efficacy. PMID- 11429174 TI - Ancient DNA from Ascaris: extraction amplification and sequences from eggs collected in coprolites. AB - On the Middle-Age site of Namur (Belgium) the analysis of coprolites revealed the presence of many well-preserved Ascaris eggs. Following rehydratation of the coprolite samples, 104 eggs were collected and extracted with an ultrasonication and phenol-chloroform based method. Three overlapping fragments of the 18S rRNA gene and one fragment of the cytochrome b gene have been reproducibly amplified, cloned and sequenced. The analysis of these sequences confirms the identification of the eggs as coming from Ascaris. Our study reveals that coprolites can be an interesting source of parasites that can be readily identified using molecular approaches. The study of ancient DNA from helminth parasites is of interest as it may answer long-standing questions in the history of infectious diseases and gives a possibility to compare these ancient sequences with those of modern populations. PMID- 11429175 TI - Complete nucleotide sequence of the 6 kb element and conserved cytochrome b gene sequences among Indian isolates of Plasmodium falciparum. AB - The malaria parasite contains a nuclear genome with 14 chromosomes and two extrachromosomal DNA molecules of 6 kb and 35 kb in size. The smallest genome, known as the 6 kb element or mitochondrial DNA, has been sequenced from several Plasmodium falciparum isolates because this is a potential drug target. Here we describe the complete nucleotide sequence of this element from an Indian isolate of P. falciparum. It is 5967 bp in size and shows 99.6% homology with the 6 kb element of other isolates. The element contains three open reading frames for mitochondrial proteins-cytochrome oxidase subunit I (CoI), subunit III (CoIII) and cytochrome b (Cyb) which were found to be expressed during blood stages of the parasite. We have also sequenced the entire cyb gene from several Indian isolates of P. falciparum. The rate of mutation in this gene was very low since 12 of 14 isolates showed the identical sequence. Only one isolate showed a maximum change in five amino acids whereas the other isolate showed only one amino acid change. However, none of the Indian isolates showed any change in those amino acids of cyb which are associated with resistance to various drugs as these drugs are not yet commonly used in India. PMID- 11429176 TI - Membrane potential changes after infection of monocytes by Toxoplasma gondii. AB - Membrane potential changes in host cell plasma membrane were analyzed and the parasitophorous vacuole membrane (PVM) potential was characterized after infection by Toxoplasma gondii. Human monocytes infested by T. gondii were stained with two membrane potential sensitive dyes, DiOC(6)(3) carbocyanine and DiSBAC(2)(3) bis-oxonol, before fluorescence emission analysis by confocal laser scanning microscopy. After 24 and 48 h of infection, 34 and 39%, respectively, of monocytes showed several parasites (from two to six) per cell. At these infection times, significant decreases in cytoplasmic emissions were observed for both DiOC(6)(3) and DiSBAC(2)(3). Thus, hyperpolarisation of the host plasma membrane would occur consecutively to infection. Inside the parasitophorous vacuole, the fluorescence intensity of DiOC(6)(3) and DiSBAC(2)(3) increased significantly from 6 to 24 h after infection and the PVM became less polarised. Involvement of different ATPases in the membrane potential of infected monocytes was evaluated with ouabain, DCCD, omeprazole and sodium orthovanadate, ATPase inhibitors. All inhibitors induced a depolarisation of the plasma membrane. In the parasitophorous vacuole compartment, DCCD, omeprazole and sodium orthovanadate but not ouabain caused a significant depolarisation of the PVM, suggesting that H(+), H(+)/K(+) and P-type ATPases were at the origin of the PVM potential. This is the first report showing the presence of ion transporters in the T. gondii PVM and the existence of at least two members of the P-type family of ion pumps: an electrogenic H(+)ATPase and an electroneutral H(+)/K(+) ATPase. PMID- 11429177 TI - Phylogenetic analysis of European species of Proteocephalus (Cestoda: Proteocephalidea): compatibility of molecular and morphological data, and parasite-host coevolution. AB - The phylogeny of European species of the tapeworm genus Proteocephalus was studied, based on partial 18S rDNA and morphological data. The group was found to be monophyletic. The analysis showed a low informative value of available morphological characters in comparison with molecular data. The morphological matrix resulted in a poorly resolved tree which is, however, compatible with the topology (Proteocephalus osculatus (Proteocephalus torulosus (Proteocephalus macrocephalus, Proteocephalus filicollis) (Proteocephalus tetrastomus, Proteocephalus percae, Proteocephalus longicollis))) based on the 18S rDNA data. A comparison performed by the program TreeMap showed a lack of significant congruency between parasite and host phylogenies. Therefore, the distribution of species in their hosts appears to be independent of the phylogeny and it is likely to be a result of host-switching, rather than co-speciation events. PMID- 11429178 TI - Effect of xanthurenic acid on infectivity of Plasmodium falciparum to Anopheles stephensi. AB - Terminally differentiated malarial gametocytes remain in the vertebrate circulation in a developmentally arrested state until they are taken up by the mosquito. The gametocytes then undergo gametogenesis in the mosquito mid-gut within minutes after ingestion of the infected blood meal. The male gametogenesis (exflagellation) can be triggered by the combination of a decrease in temperature of at least 5 degrees C and a simultaneous increase in pH between 8.0 and 8.3. Xanthurenic acid, which is present in mosquito mid-gut as well as in mosquito head, had been shown to induce exflagellation in vitro at a non-permissible pH. Here we report for the first time that with the increasing concentration of exogenous xanthurenic acid, there is a gradual increase in the number of oocysts in the mid-gut of infected mosquitoes. The concentration of xanthurenic acid for optimum infection in the membrane feeding assay was determined to be 100 microM. Three different strains of Plasmodium falciparum, viz. 3D7, 7G8 and W2 were tested in different experiments and similar findings hold true for all of them. These results demonstrate that xanthurenic acid not only induces exflagellation of male gametocytes but also promotes infectivity of Plasmodium falciparum to mosquito vectors. PMID- 11429179 TI - Treatment with beta-cyclodextrin of natural Cryptosporidium parvum infections in lambs under field conditions. AB - Following the unexpected activity of the excipient beta-cyclodextrin against experimental infection by Cryptosporidium parvum in suckling mice, its efficacy in the prevention and treatment of natural infections in lambs was evaluated under field conditions. Fifty-three crossbred neonatal lambs were randomly selected for the study. Treatment consisted of oral administration of an aqueous suspension of beta-cyclodextrin at a dose of 500 mg/kg of body weight. To test prophylactic efficacy, the suspension was administered at 1, 2 and 3 days of age. To evaluate therapeutic efficacy, the suspension was administered on each of the 3 days following onset of diarrhoea. Infection was monitored by daily examination of faecal samples, from birth to 30 days. The criteria studied in evaluating efficacy were: oocyst shedding, the presence of diarrhoea, and weight gain at 15 and 30 days. In the group that received prophylactic treatment with beta cyclodextrin, there were no mortalities and, compared with control lambs, there was a decrease in the number of animals infected, a longer prepatent period and notable reduction in the patent period and the duration of diarrhoea. Therapeutic treatment also reduced the patent period and the severity of diarrhoea. beta cyclodextrin was well tolerated by all of the treated animals. PMID- 11429180 TI - Amplified fragment length polymorphism analysis of genetic diversity of Haemonchus contortus during selection for drug resistance. AB - For the first time we used amplified fragment length polymorphism on individual nematode parasites to analyse the genetic diversity between and within isolates during consecutive stages of increased benzimidazole resistance and of increased levamisole resistance of Haemonchus contortus. The genetic diversity of the H. contortus genome turned out to be unusually high, within and between the isolates. The difference between individuals of an isolate could be as high as between individuals of two different mammalian species that do not interbreed. During benzimidazole selection the genetic constitution of the population was changed, but did not lead to a decrease in the genetic diversity. The selection for levamisole resistance resulted in a limited reduction of the genetic diversity only after the first selection step. The extensive genetic diversity apparently has allowed a fast and flexible response of H. contortus to drug selection as shown by the appearance of drug resistant isolates. This selection however has little or no effect on the extent of the genetic diversity of these resistant isolates. Implications for more sustainable control methods are discussed. PMID- 11429181 TI - Immunohistochemical and ultrastructural evidence for Neospora caninum tissue cysts in skeletal muscles of naturally infected dogs and cattle. AB - Protozoan stages were detected in the skeletal muscles of four dogs suffering from neosporosis and two neonatal calves with confirmed Neospora caninum- infection which could be immunohistochemically labelled by an antiserum against the bradyzoite-specific antigen BAG-5. In one calf, a tissue cyst was labelled by an antiserum against the N. caninum isolate NC-1. Ultrastructurally, a 0.3-1 microm-thick cyst wall surrounded the labelled parasites. The cysts were located within myofibres and contained varying numbers of bradyzoites each measuring 5.2(+/-0.6) x 1.6(+/-0.3) microm. The encysted stages showed typical ultrastructural features of N. caninum bradyzoites with subterminal nuclei, electron-dense rhoptries, and micronemes that were orientated perpendicular to the zoite pellicle. Immunohistochemistry and serology did not reveal any evidence for co-infection with Toxoplasma gondii. The detection of tissue cysts in skeletal muscle of N. caninum-infected intermediate hosts is of major epidemiological importance for the understanding of how definitive or intermediate carnivorous hosts become infected with N. caninum. PMID- 11429183 TI - 2H magnetic relaxation of 2H2O absorbed by wool. AB - D2O absorbed by intact wool fibers was studied by solid-state 2H nuclear magnetic resonance (NMR) spectroscopy. In wool fibers swollen in D2O, the deuteron transverse magnetization and the spin-locked magnetization revealed a non exponential decay. At least two NMR phases with different sets of the NMR relaxation parameters, T(1rho) (2H) and T2 2H, have been detected that may be a manifestation of two different morphological phases of the cortex of the fiber. PMID- 11429184 TI - Large solutes induce structural perturbations in proteins and membranes. AB - Structural perturbations in biopolymers with hydrophobic interiors i.e. specific proteins and dimyristoylphosphatidylcholine (DMPC) vesicles were investigated as a function of solute concentrations in the medium. 1,6-diphenyl-1,3,5-hexatriene (DPH) was used as fluorescent probe. Response of DPH was comparable to that of intrinsic tryptophan in BSA in terms of steady state and time resolved fluorescence. The solutes induced a decrease in steady state anisotropy as well as rotational correlation time (computed from lifetime measurements) for DPH in both proteins and membranes. Enhanced access of the quencher potassium iodide to tryptophan in bovine serum albumin (BSA) and ovalbumin, and enhanced terbium leakage in DMPC vesicles induced by various solutes concomitant with decreased anisotropy/correlation time were consistent with structural perturbations of the nature of defects or voids in these polymers. PMID- 11429185 TI - Development of a novel polycationic adsorbent for cryogel removal. AB - Cryogel, prevalent in the plasma of rheumatism patients, is a plasma fibronectin (pFN)-extra domain A containing FN (EDA(+)FN)-fibrinogen (Fbg) complex formed by adding heparin (HP) at a low temperature (4 degrees C). Although EDA(+)FN does not usually exist in normal plasma, its prevalence in rheumatic patients causes cryogelation in plasma. Removal of cryogel is thus a promising and novel approach to treating rheumatism. As HP-EDA(+)FN aggregate, which is induced by the main component of cryogel, is considered to be an anion, cationic materials capable of eliminating this anionic conjugate were innovated in this study. We found that an amino group density of 100-130 micromol/g (dry weight) of adsorbents prompted selective adsorption of the EDA(+)FN-HP complex. Elimination of EDA(+)FN as high as 80% accompanied by removal of the components of total FN (pFN) (10%) and Fbg (10%) in the model patient plasma was established. PMID- 11429186 TI - Role of medium--and long-range interactions in discriminating globular and membrane proteins. AB - The analysis of inter-residue interactions in protein structures provides considerable insight to understand their folding and stability. We have previously analyzed the role of medium- and long-range interactions in the folding of globular proteins. In this work, we study the distinct role of such interactions in the three-dimensional structures of membrane proteins. We observed a higher number of long-range contacts in the termini of transmembrane helical (TMH) segments, implying their role in the stabilization of helix-helix interactions. The transmembrane strand (TMS) proteins are having appreciably higher long-range contacts than that in all-beta class of globular proteins, indicating closer packing of the strands in TMS proteins. The residues in membrane spanning segments of TMH proteins have 1.3 times higher medium-range contacts than long-range contacts whereas that of TMS proteins have 14 times higher long-range contacts than medium-range contacts. Residue-wise analysis indicates that in TMH proteins, the residues Cys, Glu, Gly, Pro, Gln, Ser and Tyr have higher long-range contacts than medium-range contacts in contrast with all alpha class of globular proteins. The charged residue pairs have higher medium range contacts in all-alpha proteins, whereas hydrophobic residue pairs are dominant in TMH proteins. The information on the preference of residue pairs to form medium-range contacts has been successfully used to discriminate the TMH proteins from all-alpha proteins. The statistical significance of the results obtained from the present study has been verified using randomized structures of TMH and TMS protein templates. PMID- 11429187 TI - Composition and effect of salt on rheological and gelation properties of Enterolobium contortisilliquum gum exudate. AB - The composition, structure and rheological properties and metal ions interaction with Enterolobium contortisilliquum gum were investigated. This gum contains galactose, arabinose, rhamnose and glucuronic acid as main monosaccharide components. 13C nuclear magnetic resonance spectroscopy revealed that the anomeric composition is similar to the Enterolobium cyclocarpum exudate, however no 4-O-methylglucuronic acid was detected for E. contortisilliquum. The rheological experiment resulted in a very similar flow behaviour to that found for E. cyclocarpum gum. The empirical stiffness parameter, B was determined (B=0.090) and suggested that the polysaccharide possesses semi-flexible chains. Cation affinity of the gum aqueous solution was determined by intrinsic viscosity measurements and gelation. The melting temperature of gels pointed to a charge/ionic radius ratio dependence for metal ions. Thermodynamic parameters (enthalpy and entropy) at the melting temperature were also calculated. PMID- 11429188 TI - Influence of pectins on the solubility and the molar mass distribution of dehydrogenative polymers (DHPs, lignin model compounds). AB - Dehydrogenation polymers (DHPs, lignin model compounds) were synthesized in the presence of increasing pectin concentrations using two different methods. The first method ('Zutropfverfahren', ZT) consists in the slow adding of monomers whereas in the second method ('Zulaufverfahren', ZL) all the reactants are added simultaneously. DHPs solubility increases with the pectin concentration in the ZT experiments and remains stable in the ZL experiments. Covalent bonds between pectin and DHP are formed during ZT polymerization resulting in lignin carbohydrate complex (LCC) which keeps the unbound DHPs in solution by the formation of aggregate or micelle-like structures. In contrast LCC are not formed during the ZL process which behave like the DHP reference. The ZT DHP molar masses increase observed is attributed to the reactivity of the high molar mass polymer solubilized by the LCC whereas ZL higher molar mass polymers are precipitated out of the solution and cannot react further. PMID- 11429189 TI - Shear creep of gelatin gels from mammalian and piscine collagens. AB - This paper describes new measurements on the creep rheological behaviour of gelatin gels from both traditional mammalian and piscine sources. Measurements on a series of concentrations of gels were obtained using a high-precision controlled stress rheometer. Results for the concentration dependence of compliance are close to those expected from dynamic oscillatory measurements of gel modulus, assuming ideal elasticity. The concentration dependence of viscosity approximates power law behaviour, with eta~C( approximately 2-3), lower than the exponent expected for semi-dilute solutions. The apparent contradiction implied by this is discussed and a novel gel viscosity versus concentration state diagram presented. PMID- 11429190 TI - Use of RT-PCR to detect co-expression of neuropeptides and their receptors in lung cancer. AB - Small-cell lung cancer (SCLC) synthesises a wide range of neuropeptides and their corresponding receptors. Together, these can form autocrine growth loops. Non small-cell lung cancer (NSCLC) does not generally share this neuroendocrine phenotype. In this study, we tested the hypothesis that multiple neuropeptides and their receptors are co-expressed in SCLC, constituting potential autocrine loops. Expression of mRNA for arginine vasopressin, gastrin, cholecystokinin, gastrin-releasing peptide, endothelin and neurotensin, together with their cognate receptors, was evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) in a panel of human lung cancer cell lines. We have assessed those neuropeptides and neuropeptide receptors that could be used as potential early markers to detect lung cancer cells both as micrometastases in blood and within dysplasia in bronchial biopsies. We establish that although no cell line expressed all neuropeptides, co-expression of neuropeptides and their receptors is common in SCLC but not in NSCLC. We conclude that mRNA for the neuropeptides gastrin-releasing peptide and arginine vasopressin and the cholecystokinin receptor B were most SCLC-specific and RT-PCR for these markers could be used to distinguish between SCLC and NSCLC. PMID- 11429191 TI - Identification of small lung nodules at autopsy: implications for lung cancer screening and overdiagnosis bias. AB - PURPOSE: Unsuspected cases of lung cancer are reported to be uncommon in autopsy series, and these data have been used to suggest that indolent tumors are rare and that overdiagnosis bias is not an important factor in lung cancer screening. The purpose of this study was to determine if a retrospective autopsy review is indeed accurate in identifying all small lung nodules on CT, and thus provide a true estimate of unsuspected lung tumors. MATERIALS AND METHODS: We identified all 1047 patients who had an autopsy at our institution from 1994 to 1998. We then reviewed the patients radiology records and found 187 patients with a thoracic CT within 2 months of the postmortem examination. All 187 CT reports were reviewed in order to identify patients with at least one pulmonary nodule. CT studies with reports that described a nodule(s) were then re-reviewed to confirm presence and location of the nodule(s). The CT findings were than compared to the autopsy report to determine if the postmortem examination indeed found the nodule(s). RESULTS: 28 autopsy patients had at least one pulmonary nodule identified on their thoracic CT no more than 2 months before death. Nineteen patients (68%) had nodule(s) recorded on the autopsy report, two ( approximately 10%) of which proved to have undiagnosed squamous cell carcinoma. Nine patients (22%) had no mention of pulmonary nodules seen on the CT recorded on their autopsy report. CONCLUSIONS: This study suggests autopsies do not identify all small pulmonary nodules found at CT. The true incidence of clinically insignificant lung cancer is thus uncertain, and overdiagnosis bias in lung cancer screening may be more important than previously recognized. PMID- 11429192 TI - Prognostic value of ground-glass opacity found in small lung adenocarcinoma on high-resolution CT scanning. AB - OBJECTIVE: This study was undertaken to investigate the value of the ground-glass opacity (GGO) area found on high-resolution computed tomography (HRCT) scanning as a preoperative prognostic indicator. PATIENTS AND METHODS: We studied 104 patients with small-sized lung adenocarcinoma, 20 mm or less in diameter, between 1995 and 1999. Three independent radiologists semi-quantitatively scored the extent of GGO on HRCT as greater than or less than 50%. Three independent pathologists semi-quantitatively scored the extent of the bronchioloalveolar carcinoma (BAC) component of the tumor on histologic examination as greater than or less than 50%. As no relapse occurred in patients with GGO greater than 50%, multivariate analysis of this prognostic factor was not possible. RESULTS: Fifty patients were scored as having both BAC and GGO greater than 50%, 36 as both BAC and GGO less than 50%, and 16 as BAC greater than 50% and GGO less than 50%. In only two patients (1.9%), BAC less than 50% was overestimated on HRCT as GGO greater than 50%. The sensitivity and specificity of GGO to BAC were 76 and 95%, respectively. The 3 year-relapse-free survival rates in each group of 52 patients with GGO greater than and less than 50% were 100 and 72%, respectively, after a median follow-up of 24 months. Univariate analysis indicated that both GGO and BAC areas were significantly correlated with cancer relapse (P=0.005 and P=0.002). The multivariate analysis revealed an independent prognostic influence of the BAC area on relapse-free survival (P=0.015, relative risk=0.07). CONCLUSIONS: To date there has been no relapse among the 52 patients with GGO greater than 50%. This novel classification based on the semiquantitative analysis of GGO area on HRCT should become an useful independent preoperative indicator when deciding on operative procedure, and to predict the potential of relapse in patients with small adenocarcinoma arising from the peripheral lung. PMID- 11429193 TI - Interaction between novel anticancer agents and radiation in non-small cell lung cancer cell lines. AB - Integration of chemotherapy and radiation is the standard practice in the management of locally advanced inoperable NSCLC. To assess the biological interaction between third generation chemotherapeutic agents and radiation in non small cell lung cancer (NSCLC) in vitro, we tested a number of different drugs (paclitaxel, docetaxel, gemcitabine, topotecan, SN-38 and cisplatin) combined with radiation, in lung cancer cell lines. Cellular chemosensitivity was determined, using the semi-automated colorimetric MTT assay, after 48, 72 and 96 h of exposure to increasing drug concentrations, (0.001-100 microM) and radiation doses (100-400 cGy). Cell lines used were the adenocarcinoma (ADK), A-549, and the squamous-cell carcinoma (SCC), LX-1. Cells were pre-treated with anticancer agents at 24, 12 and 0 h before irradiation. Cytofluorimetric cell cycle analysis was performed. A significant S-phase block or a G(2)/M block was seen with gemcitabine and topotecan or paclitaxel pre-treatment, respectively. Apoptosis was seen only after paclitaxel exposure in the A-549 cell line. Despite a similar pattern of cell-kinetic changes induced by chemotherapy pre-treatment in all cell lines, the adenocarcinoma A-549 cell line was not radiosensitized by any of the anticancer agents tested, whereas synergism was observed in the LX-1 squamous carcinoma cell line, when exposed to gemcitabine, SN-38, topotecan and cisplatin. Paclitaxel, despite a favourable cell cycle effect, was not found to be synergistic with radiotherapy in our experimental model. In conclusion, the observed synergism appears to be dose- and timing-independent and seems to be related to the histological subtype being present in SCC only. Favourable perturbation of the cell cycle is evident with all the new agents tested in both cell types, but was not sufficient to produce synergism with radiation. PMID- 11429194 TI - A comparative study of telomerase activity in sputum, bronchial washing and biopsy specimens of lung cancer. AB - The potential of telomerase, the ribonucleoprotein enzyme, as a non-invasive screening marker was studied in pre-bronchoscopy sputum (S), bronchial washings (W) and bronchoscopic biopsy (B) samples from individuals under evaluation for lung cancer. Out of the 52 cases studied, 42 were clinically suspected primary lung cancer patients and 10 had pulmonary disorders but had no clinical evidence of lung cancer. Fifteen (39.5%) S samples, 24 (63.1%) W samples and 32 (84.2%) B samples, which were cytologically/histopathologically positive were also positive for telomerase activity. Interestingly, 16 (42%) S samples, 20 (52.6%) W samples and 20 (52.6%) B samples, initially reported cytologically/histopathologically negative, showed detectable telomerase activity. Lung cancer was finally confirmed in these cases by repeat cytology/histopathology. However, telomerase activity was detected in 31 (81.6%) S, 26 (68.4%) W and 33 (86.8%) B samples of suspected lung cancer patients. Telomerase activity was negative in S, W, and B of four of the suspected cases, which ultimately turned out to be negative for lung cancer. Cytopathology/histopathology alone (including repeat attempts) identified 15 (39.5%) cases of sputum, 24 (63.1%) cases of bronchial washings and 32 (84.2%) bronchoscopic biopsy samples. Out of 10 controls, low telomerase activity was detected in only one (10%) of the bronchial washings, which later turned out to be due to large number of inflammatory cells. Telomerase activity assay of sputum carried sensitivity, specificity and diagnostic accuracy of 81.6, 100 and 86.5%, respectively, while that for bronchial washing was 68.4, 100 and 76.9%, respectively, and for bronchoscopic biopsy samples was 86.8, 100 and 88.1%, respectively. A positive correlation (P<0.01) was seen between age and telomerase activity in sputum, bronchial washing and biopsy samples but no significant correlation was seen between sex and telomerase activity or duration of smoking and telomerase activity. A significant positive correlation was observed between staging and telomerase activity in sputum (P<0.01), bronchial washing (P<0.01) and biopsy samples (P<0.01). Our findings indicate that telomerase is a specific marker for malignant lung disease and can complement cytology/histopathology in the diagnosis of lung cancer. Sputum telomerase assay holds the potential for early and non-invasive diagnosis of lung cancer. PMID- 11429195 TI - Early hilar lung cancer--risk for multiple lung cancers and clinical outcome. AB - In early hilar lung cancer patients, multiple lung cancers frequently develop. The clinical outcome of such patients were studied. A total of 91 patients, 88 men and three women, who were endoscopically diagnosed with early hilar lung cancer were studied retrospectively. Surgery was performed in 46 patients, while organ-sparing treatment, including photodynamic therapy (PDT), Nd-YAG (neodymium yttrium, argon, garnet) laser vaporization, and radiotherapy, were done for 45 patients. During follow-up, newly developed lung cancers and/or malignancies in other organs were recorded. The average smoking index (cigarettes per day x years) was 1040. Synchronous and/or metachronous multiple lung cancers developed in 26/91 patients (28.6%). Malignancies in other organs were found in 12/91 (13.2%). The smoking index of patients with multiple lung cancers was significantly higher than for other patients. The overall 5 year survival rate was 70.7% in all patients, 76.0% in the surgery group, and 64.4% in the nonsurgery group. The lung cancer-specific 5 year survival rate was 89.8% in all patients, 89.3% in the surgery group, and 90.5% in the nonsurgery group. Early hilar lung cancer frequently accompanies other lung cancers or malignancies in other organs. A favorable prognosis can be obtained with organ-sparing treatment. PMID- 11429196 TI - Increased expression of high but not low molecular weight heat shock proteins in resectable lung carcinoma. AB - Strong expression of high-molecular-weight (HMW) heat-shock proteins (HSP) by lung carcinoma has been documented using immunohistochemistry. Far less is known about the expression of low-molecular-weight (LMW) HSP in lung cancer. We compared the quantitative expression of HMW (HSP-60, HSP-70) and LMW (HSP-27, ubiquitin) HSP in tumor and non-tumor lung tissue obtained from 47 patients undergoing surgical resection of lung carcinoma. HSP levels were determined in cell lysates from tissue samples by ELISA using streptavidin-biotin technology. Results were normalized to total protein content measured by spectrophotometry. Compared to disease-free lung tissue, tumor tissue samples showed higher levels of both HSP-60 (median value: 227 pg versus 96 pg per mg protein (P<0.001 by Wilcoxon Rank test for paired data) and HSP-70 (median value: 525 ng versus 401 ng per mg protein (P=0.01 by Wilcoxon Rank test for paired data). Tumor and tumor free tissues show similar levels of ubiquitin and HSP-27. Neither the survival rate nor the histologic type and extent of cancer are correlated with the observed differences in HSP-60 and HSP-70 expression (P>0.1 by one way analysis of variance for repeated measures with one between subject factor). Our data confirm, on a quantitative basis, the increased expression of HSP-60 and HSP-70 in non-small-cell lung carcinoma. However, no prognostic value was found to be associated with this over-expression. In contrast, LMW stress proteins such as ubiquitin and HSP-27, although implicated in cellular processes potentially related to malignant transformation, show no increased expression in lung carcinoma. PMID- 11429197 TI - A phase I study of weekly docetaxel and cisplatin in advanced non-small cell lung cancer. AB - This phase I study was designed to determine the maximum tolerated dose (MTD) and toxicity of a weekly docetaxel (TXT) and cisplatin (CDDP) combination regimen in advanced non-small cell lung cancer (NSCLC). Patients with stage IIIB or IV NSCLC who were previously untreated were eligible. Docetaxel, at a starting dose of 20 mg m(-2) per week on days 1, 8 and 15, was combined with a fixed dose of cisplatin 80 mg m(-2) on day 1. Docetaxel was increased in 5 mg m(-2) per week steps. Chemotherapy was given in a 4-weeks cycle. Dose-limiting toxicities (DLTs) were defined as grade 3-4 leukopenia, thrombocytopenia, anemia, fever with grade 4 neutropenia and more than grade 2 non-hematologic toxicity, with the exception of nausea, vomiting, and alopecia. Omission of chemotherapy on day 8 and/or 15 was also considered DLT. Eighteen patients were enrolled in this study. Leukopenia, anemia and fatigue were the DLTs. No grade 4 toxicities were seen in any patients. The overall response rate was 44.4% (95% confidence interval, 21.5 67.4%). The recommended dose of TXT to be combined with CDDP 80 mg m(-2) on day 1 is 35 mg m(-2) per week on days 1, 8 and 15. This is a promising regimen, therefore a multicenter phase II study is now under way. PMID- 11429198 TI - Endoscopic argon plasma coagulation for palliative treatment of malignant airway obstructions: early results in 47 cases. AB - Argon plasma coagulation (APC) is a new method of non-contact electrocoagulation, using high frequency current by means of ionized argon gas (argon plasma). Recently, this technique has become available for flexible endoscopic delivery through special probes. Aim of this study is to evaluate the efficacy, indications and the possible side effects of APC use in the palliative treatment of malignant airway obstructions and/or bleeding. Over a 24-month period, 47 patients underwent APC treatment for malignant neoplasms of the tracheobronchial system causing obstruction and/or recurrent bleeding. Immediate airway patency and haemostasis were obtained in 91.5% of cases (43/47). No complications or side effects caused by the treatment were observed. In two patients, the treatment allowed a radical surgical approach after induction chemotherapy. In all cases, APC proved to be highly effective and easy to perform. In our experience, APC has proven to be easy to perform, rapidly effective, safe and well tolerated by the patient, even after repeated application. This study highlights the value of endoscopic APC in the palliative management of tracheobronchial neoplasms. PMID- 11429199 TI - Lung cancer as the presenting feature of AIDS. AB - Several reports have suggested that there is an increased incidence of lung cancer amongst the HIV positive population. All cases of lung cancer reported in this group to date were in patients known previously to be HIV positive. We now report a case of AIDS presenting with small cell lung cancer. Unexpectedly severe pancytopenia and immunosuppression after combination chemotherapy revealed the underlying diagnosis. The prognosis in such cases is very poor, but it is important to establish the correct diagnosis in order for patients to make informed treatment decisions and to allow potentially life-saving treatment of their sexual partners. PMID- 11429200 TI - Furanochromone radical cations: generation, characterization and interaction with DNA. AB - The radical cations of naturally occurring furanochromones visnagin (VI) and khellin (KH) have been generated and identified for the first time by use of laser flash photolysis and pulse radiolysis techniques. The lifetimes of VI(.+) and KH(.+) are determined as approximately 6 and approximately 35 micros under these conditions, respectively. Direct 308-nm excitation of VI in aqueous buffer at physiological pH results in monophotonic photoionization to generate VI(.+), with a quantum yield of 0.075, which is much higher than that of 8 methoxypsoralen and KH under identical conditions. Though VI(.+) is a more powerful oxidant than KH(.+), both of them react with guanosine mononucleotide (k=1.2x10(9) and 3.8x10(7) dm(3) mol(-1) s(-1), respectively) via electron transfer to give the guanine radical cation. Furthermore, selective oxidation of guanine in single and double strand DNA by VI(.+) was also observed. These novel findings suggest that electron transfer reactions involving furanochromone radical cations may be of considerable importance in furanochromone photochemotherapy. PMID- 11429201 TI - Analysis of peptides synthesized in the presence of SAz-1 montmorillonite and Cu(2+) exchanged hectorite. AB - We have investigated the synthesis of oligopeptides containing glycine and tyrosine in the presence of the clay minerals montmorillonite (non-exchanged, SAz 1) and Cu(2+) exchanged hectorite. In both cases, homopolymers of the two amino acids are formed, as are mixed peptides. In the case of Cu(2+) hectorite, mixed oligopeptides up to trimers are detected in small amounts. For montmorillonite, heterogeneous oligopeptides up to hexamers are detected. Our experiments indicate montmorillonite is more effective in promoting oligopeptide formation than Cu(2+) hectorite. Analysis of the oligopeptide sequences formed on the montmorillonite surfaces indicates preferential synthesis of certain Gly-Tyr sequences over others. PMID- 11429202 TI - The interactions of amphotericin B with various sterols in relation to its possible use in anticancer therapy. AB - Amphotericin B (AmB) is still the most common anti-fungal agent used to treat systemic fungal infections. It is known that this antibiotic acts by forming pores with the ergosterol contained in the membranes of fungi, but it also interacts with the cholesterol contained in the membranes of eukaryotic cells, hence its toxicity. AmB may also interact with the most common oxidation products of cholesterol found in vivo, together with interacting with biosynthetic precursors of cholesterol, namely, lanosterol and 7-dehydrocholesterol (7-DHC). The purpose of the present work was to study the interactions in solution between AmB and these various sterols, the techniques used being UV-Vis spectroscopy and differential scanning calorimetry. The results are globally interpreted in terms of the structural differences between the sterols. We show that AmB selectively interacts with 7-DHC which, according to a recent hypothesis proposed in the literature, has been identified in connexion with a therapeutic strategy against hepatocellular carcinomas. We find that the affinity of AmB towards 7-DHC is even greater than the affinity of the antibiotic towards ergosterol. We also find that AmB selectively interacts with the principal oxidation product of cholesterol, 7 ketocholesterol, a situation that has to be taken into account when AmB is administered. PMID- 11429203 TI - Influence of N6-isopentenyladenosine (i(6)A) on thermal stability of RNA duplexes. AB - The thermodynamic stability of self-complementary oligoribonucleotides containing N6-isopentenyladenosine (i(6)A) or N6-isopentanyladenosine (p(6)A) was determined. The base pairs i(6)A.U and p(6)A.U were placed in either an internal (separated and tandem) and a terminal position within the duplex, or unpaired i(6)A and p(6)A as a 3'-dangling ends. The thermal unfolding of the oligomers was determined by means of UV melting profiles and the thermodynamic parameters: enthalpy (DeltaH degrees ), entropy (DeltaS degrees) and free energy (DeltaG degrees (37)) as well as the melting temperature (T(m)) were calculated. Both modified nucleosides destabilized the duplexes, however, the effect depended on the position of the modified adenosine within the duplex. The similarity of the behavior of oligomers containing i(6)A and p(6)A suggests a negligible effect of the double bond on the thermal stability. The largest destabilization was observed when derivatives of adenosine were placed in an internal position. The effect of 3'-dangling ends suggests that the presence of the N6-isopentenyl- or N6-isopentanyl substitutent affects hydrogen bonding rather than stacking within duplex. PMID- 11429204 TI - Study of human serum albumin-TiO(2) nanocrystalline electrodes interaction by impedance electrochemical spectroscopy. AB - The adsorption of human serum albumin (HSA) onto nanocrystalline TiO(2) electrodes was studied by electrochemical impedance spectroscopy (EIS) in function of pH and electrode potential. The characterization and physico-chemical properties of the TiO(2) electrode were investigated by scanning electron microscopy (SEM), UV-photoelectron spectroscopy (UPS), cyclic voltammetry and capacitance measurements. The impedance response of the particulate TiO(2) electrode/protein interface was fitted using an equivalent circuit model to describe the adsorption process. The adsorbed protein layer, which is formed as soon as the protein is injected into the solution and becomes in contact with the electrode, was investigated as a function of electrode potential and solution pH. The measurements were performed under pseudo-steady-state and steady-state conditions, which gave information about the different states of the system. With the pseudo-steady state measurements, it was possible to determine two rate constants of the protein adsorption process, which correspond to two different states of the protein. The shortest one was associated with the first contact between the protein and the substrate and the second relaxation time, with the protein suffering an structural rearrangement due to the interaction with the TiO(2) electrode. It was detected that at sufficiently long times (approx. 1 h, where the system was under steady state conditions), a quasi-reversible protein adsorption mechanism was established. The measurements performed as a function of frequency under steady-state conditions, an equivalent circuit with a Warburg element gave the better fitting to data taken at -0.585 V closer to the oxide flat band potential and it was associated with protein diffusion. Experimental results obtained at only one frequency as a function of potential could be fitted to a model that takes into account non-specific and probable specific protein adsorption, which renders to be potential- and pH-dependent. Low capacity values were obtained in the whole potential range, which were measured in the presence and in the absence of the protein layer. The capacity dependence on potential and pH were associated with the generation of surface states on TiO(2). A surface state concentration of 4.1x10(18) cm(-2) was obtained by relating the parallel capacitance with oxide surface states arising from the protein-oxide interaction. PMID- 11429205 TI - Noise enhanced hormonal signal transduction through intracellular calcium oscillations. AB - In a wide range of non-linear dynamical systems, noise may enhance the detection of weak deterministic input signals. Here, we demonstrate this phenomenon for transmembrane signaling in a hormonal model system of intracellular Ca(2+) oscillations. Adding Gaussian noise to a subthreshold extracellular pulsatile stimulus increased the sensitivity in the dose-response relation of the Ca(2+) oscillations compared to the same noise signal added as a constant mean level. These findings may have important physiological consequences for the operation of hormonal and other physiological signal transduction systems close to the threshold level. PMID- 11429206 TI - Evidence for carbon monoxide binding to sickle cell polymers during melting. AB - The melting of sickle cell hemoglobin (HbS) polymers was induced by rapid dilution using a stopped-flow apparatus. The kinetics of polymer melting were monitored using light scattering. Polymer melting in the absence of any hemoglobin ligand was compared to melting when the diluting buffer was saturated with carbon monoxide (CO). In this way the role of CO in polymer melting could be assessed. The data were analyzed using models that assumed that melting occurs only at the ends of polymers. It was further assumed that CO could only bind to HbS in the solution phase. However, our data could not be fitted to this model, where CO cannot bind directly to the polymer. Thus, CO probably binds directly to the polymers during our melting experiments. This result is discussed in terms of oxygen induced polymer melting and polymerization processes in sickle cell disease PMID- 11429207 TI - Influence of the oligomeric state of yeast hexokinase isozymes on inactivation and unfolding by urea. AB - The effect of the association-dissociation equilibrium on the urea-induced inactivation and unfolding of the yeast hexokinase isoforms, PI and PII, showed that these enzymes are more stable as dimers. For the monomeric PII, the inactivation and unfolding processes occurred in parallel. However, inactivation precedes the unfolding of monomeric PI or dimeric PI and PII. The unfolding transitions are biphasic for PI indicating stable intermediates, whereas for the PII isoform the unfolding occurs in a single step. Our data suggests that although PI and PII present a 78% identity in their amino acid sequences, they probably have distinct inactivation and unfolding by urea behavior. PMID- 11429208 TI - A fluorescence-based assay for transcription using a novel fluorescent GTP analogue. AB - A new fluorescent analogue of GTP (Cm-GTP) was synthesized, which contained a coumarin fluorophore attached to the gamma phosphorus. This compound was tested in transcription assays using T7 RNA polymerase as a model system. The fluorescent nucleotide was incorporated specifically at the 5' end of nascent RNA synthesized in two different modes of transcription initiation. In the first mode, with only Cm-GTP (+GTP), reiterative slippage synthesis occurred and poly rG ladders of up to 14 nucleotides were synthesized. In the second mode, with Cm GTP (+GTP)+ATP+CTP, abortive transcripts of up to seven or eight nucleotides were synthesized. The fluorescence properties of the two types of RNA were studied in detail. There was greater reduction in fluorescence intensity in G-ladders than in abortive transcripts. Steady-state anisotropy and anisotropy decay indicated that the fluorophore motion was constrained in G-ladder RNAs as compared to abortive RNAs. Quenching experiments by using extraneous quenchers showed that the excited state of fluorophore at the 5' end of G-ladder RNA was less efficiently quenched as compared to the free fluorophore. These studies suggested that the fluorescent GTP analogue sensed the structural features that distinguished G-ladder RNAs from abortive RNAs. The results suggested that G ladder RNAs adapt unusual conformations such as G-quartets. Thus, the new fluorescent probe can be useful for structural studies on RNA. PMID- 11429210 TI - Patterns and fate of PSA bouncing following 3D-CRT. AB - PURPOSE: The goals of this study were to quantify the frequency of post-treatment prostate-specific antigen (PSA)-level bouncing following three-dimensional conformal radiation therapy (3D-CRT) for prostate cancer and to identify any relationships that may exist between bouncing activity and biochemical control (bNED). METHODS: Between May 1989 and July 1995, 306 patients were treated with 3D-CRT alone. All patients had 6 or more post-treatment PSA levels and at least 5 years of PSA follow-up. The median total follow-up and total dose to the center of prostate was 79 months and 74 Gy, respectively. A bounce was defined by a minimum rise in PSA of 0.4 ng/mL over a 6-month period, followed by a drop in PSA of any magnitude. Estimates of bNED control rates were made using Kaplan-Meier methodology and comparisons were made using the log-rank test. Multivariate analysis of bNED control predictors was accomplished using a stepwise Cox proportional hazards model. RESULTS: Nearly one third of the patients experienced at least one bounce. Bouncers were found to present with higher pretreatment PSA levels and were treated with lower dose levels to the center of prostate. Five year bNED control estimates for nonbouncers vs. bouncers were 69% and 52%, respectively (p = 0.0024). After controlling for dose and pretreatment PSA level, total number of bounces emerged as a significant predictor of bNED control (p = 0.02). CONCLUSIONS: Bouncing PSA levels occur in approximately one third of the patients treated with 3D-CRT alone, with bouncing occurring at a constant rate from 2 to 5 years post-treatment. Bouncing is associated with lower radiation dose levels, higher pretreatment PSA levels, and decreased bNED control. Nearly half of the bouncers are bNED controlled; thus, clinicians should not use bouncing as a sole indicator of relapse. PMID- 11429211 TI - Radiation-induced hypoxia may perpetuate late normal tissue injury. AB - PURPOSE: The purpose of this study was to determine whether or not hypoxia develops in rat lung tissue after radiation. METHODS AND MATERIALS: Fisher-344 rats were irradiated to the right hemithorax using a single dose of 28 Gy. Pulmonary function was assessed by measuring the changes in respiratory rate every 2 weeks, for 6 months after irradiation. The hypoxia marker was administered 3 h before euthanasia. The tissues were harvested at 6 weeks and 6 months after irradiation and processed for immunohistochemistry. RESULTS: A moderate hypoxia was detected in the rat lungs at 6 weeks after irradiation, before the onset of functional or histopathologic changes. The more severe hypoxia, that developed at the later time points (6 months) after irradiation, was associated with a significant increase in macrophage activity, collagen deposition, lung fibrosis, and elevation in the respiratory rate. Immunohistochemistry studies revealed an increase in TGF-beta, VEGF, and CD-31 endothelial cell marker, suggesting a hypoxia-mediated activation of the profibrinogenic and proangiogenic pathways. CONCLUSION: A new paradigm of radiation-induced lung injury should consider postradiation hypoxia to be an important contributing factor mediating a continuous production of a number of inflammatory and fibrogenic cytokines. PMID- 11429212 TI - Reciprocal translocations in patients with testicular seminoma before and after radiotherapy. AB - PURPOSE: The evaluation of radiation-induced chromosomal translocations in peripheral lymphocytes using fluorescent in situ hybridization is a promising method for retrospective dosimetry after a radiation accident. We evaluated the genomic frequency of chromosomal translocations in patients with testicular seminoma who received adjuvant radiotherapy to the retroperitoneal lymph nodes, to evaluate the time-effect relationship of radiation-induced stable aberrations after partial body irradiation. METHODS: In 13 patients, peripheral lymphocytes could be evaluated before radiotherapy and at several time points after radiotherapy. In 17 additional patients, lymphocyte samples were obtained after radiotherapy. Thirteen healthy men served as age-matched controls for the aberration frequency before radiotherapy. Fluorescent in situ hybridization was performed using whole chromosome probes against chromosomes No. 4, No. 6, and No. 7. RESULTS: Nearly all patients displayed an increased spontaneous rate of genomic translocations (F(G)) before radiotherapy compared to age-matched, healthy men. The difference was significant in the paired ranks test (p < 0.0001). After adjuvant radiotherapy, the F(G) increased 2- to 8-fold in individual patients. Within 20 months after radiotherapy, the F(G) returned to pretherapeutic levels. CONCLUSIONS: The frequency of genomic translocations after partial body irradiation is time dependent. A persistence of chromosomal aberrations, which is to be expected after total body irradiation, could not be observed. It is likely that the dose and the volume of the irradiated bone marrow are playing a role in the persistence of stable chromosomal aberrations. Patients with testicular seminoma displayed an increased frequency of spontaneous genomic translocations before the initiation of radiotherapy. This chromosomal instability might be related to the known increased rate of secondary cancers in this patient group. PMID- 11429213 TI - Tumor hypoxia, p53, and prognosis in cervical cancers. AB - BACKGROUND: The p53 protein is involved in the regulation of initiation of apoptosis. In vitro, p53-deficient cells do not respond to hypoxia with apoptosis as do p53-normal cells, and this may lead to a relative growth advantage of cells without a functioning p53 under hypoxia. On the basis of this hypothesis, a selection of cells with a functionally inactive p53 may occur in hypoxic tumors. The development of uterine cervical carcinomas is closely associated with infections of human papilloma viruses, which may cause a degradation of the tumor suppressor gene p53, resulting in a restriction of apoptosis. Thus, cervical cancers have often a functionally inactive p53. The purpose of our clinical study was therefore to investigate the association between p53, hypoxia, and prognosis in cervical cancers in which the oxygenation status can be determined by clinical methods. MATERIAL AND METHODS: Seventy patients with locally advanced squamous cell cervical cancer Stages IIB (n = 14), IIIB (n = 49), and IVA (n = 7) were investigated in the period from 1996 through 1999. All were treated with definitive radiotherapy with curative intent by a combination of external radiotherapy plus high-dose-rate afterloading. Before therapy, tumor oxygenation was measured with a needle probe polarographically using the Eppendorf histograph. Hypoxic tumors were defined as those with pO(2) measurements below 5 mm Hg (HF5). Pretreatment biopsies were taken and analyzed immunohistologically for p53 protein expression with the DO-7 antibody. The DNA index was measured by flow cytometry. The statistical data analysis was done with SPSS 9.0 for Windows. RESULTS: The 3-year overall survival was 55% for the whole group of patients. Clinical prognostic factors in a multivariate analysis were pretreatment hemoglobin level (3-year survival 62% for patients with a pretreatment hemoglobin > or =11 g/dl vs. 27% for hemoglobin <11 g/dl, p = 0.006) and FIGO stage (Stage IIB: 65%; Stage IIIB: 60%; Stage IVA: 29%, p = 0.01). Sixty of the 70 tumors showed positive immunohistologic staining for p53 protein (transformed p53 = tp53), and 10/70 were negative (wild-type p53 = wtp53); p53 expression had no significant impact on survival (50% for tp53 vs. 79% for wtp53, p = 0.11). FIGO stage and anemia had no impact on p53 expression. Forty-nine of 70 tumors were hypoxic (HF5+), and 21 showed no hypoxia (HF5-). Hypoxic carcinomas were more frequently positive for p53 as compared to nonhypoxic tumors (27% vs. 13%, p = 0.011) and showed a trend toward a lower survival (48% vs. 70%, p = 0.07). In a further multivariate analysis, the impact of a combination of p53 expression and hypoxia on survival was examined. After adjusting for FIGO stage and pretreatment anemia, patients with wtp53 tumors had the best prognosis (3-year survival 79%) followed by tp53-HF5(-) patients (57%), and the most unfavorable prognosis was observed for tp53-HF5(+) patients (47%). The DNA index was higher in tp53 carcinomas compared to wtp53 tumors, 1.97 +/- 0.4 vs. 1.67 +/- 0.1, p = 0.05. The highest DNA index was found in hypoxic tumors with transformed p53 (2.2 +/- 3.1). CONCLUSIONS: Advanced stage and pretreatment hemoglobin level are independent prognostic factors in cervical carcinomas. The immunohistologic detection of (a functionally inactive) p53 and the presence of hypoxia had no prognostic impact, if analyzed as single parameters. However, the combination of both parameters was able to discriminate different prognostic subgroups. Moreover, hypoxic cancers were more often immunohistologically positive for tp53 protein and had a higher DNA index with the highest DNA index in tumors with both hypoxia and tp53 protein expression. These findings in summary support the theory that the tumor's microenvironment may influence the biologic behavior via hypoxia. PMID- 11429214 TI - Combined brachytherapy and surgery for early carcinoma of the uterine cervix: analysis of extent of surgery on outcome. AB - PURPOSE: The aim of this retrospective study was to evaluate the survival data and rates and patterns of complications and recurrences for patients who had early uterine cervix carcinoma and underwent brachytherapy and subsequent surgery. METHODS AND MATERIALS: Between January 1990 and December 1997, 192 women with cervical carcinoma (Stages IA2 with vascular invasion [n = 28], IB1 [n = 144], and IIA [n = 20]) underwent brachytherapy, delivering 60 Gy and then hysterectomy with external iliac lymphadenectomy. Piver class I, II, and III hysterectomies were performed on 136, 38, and 18 patients, respectively. Adjuvant chemoradiotherapy was delivered to patients with positive lymph nodes. RESULTS: The median follow-up time was 61 months. After brachytherapy, a pathologically complete response (CR) was observed in 137 (71.3%) of 192 women. The distribution of CRs according to tumor stage was as follows: Stage IA2, 24 (85.7%) of 28; Stage IB1, 105 (72.9%) of 144; and Stage IIA, 8 (40%) of 20. Patients with Stage IB1 cancer had 13 lymph node metastases (9%), as did 6 with Stage IIA disease (30%). Pelvic recurrences occurred in 9 (4.6%) of the 192 patients; in 3, local relapses were associated with relapses at distant sites. Ten patients had systemic relapses (5.2%). Recurrences at distant sites were more frequent (p < 0.02) in partial responders, and other recurrences were more frequent in patients with lymph node metastases (p < 0.04). The overall 5-year disease-free survival rate was 91.2% (96.2% for Stage IA2, 91% for Stage IB1, and 84.4% for Stage IIA cancers). The class of hysterectomy did not influence the outcome. Late complications occurred in 28 patients (Grade 1, 24 [12.5%]; Grade 2, 4 [2%]; and Grade 3, 1 [0.5%] of 192 patients). CONCLUSIONS: Combined treatments resulted in high local control and low morbidity rates in patients with early-stage cervical carcinoma. Limited surgery seemed to be adequate after intracavitary therapy. PMID- 11429215 TI - Early toxicity from preoperative radiotherapy with continuous infusion 5 fluorouracil for resectable adenocarcinoma of the rectum: a Phase II trial for the Trans-Tasman Radiation Oncology Group. AB - PURPOSE: To assess the toxicity and the efficacy of preoperative radiotherapy with continuous infusion 5-fluorouracil (5-FU) for locally advanced adenocarcinoma of the rectum. METHODS AND MATERIALS: Eligible patients had newly diagnosed localized adenocarcinoma of the rectum within 12 cm of the anal verge, Stage T3-4, and were suitable for curative resection. Eighty-two patients were treated with radiotherapy-50.4 Gy in 28 fractions in 5.6 weeks, given concurrently with continuous infusion 5-FU, using either 96-h/week infusion at 300 mg/m(2)/day or 7-days/week infusion at 225 mg/m(2)/day. RESULTS: The median age was 59 years (range, 27-87), and 67% of patients were male. Pretreatment stages of the rectal cancer were T3, 89% and resectable T4, 11%, with endorectal ultrasound confirmation in 67% of patients. Grade 3 acute toxicity occurred in 5 of 82 patients (6%; 95% confidence interval [CI], 2-14%). Types of surgical resection were anterior resection, 61%; abdominoperineal resection, 35%; and other procedures, 4%. There was no operative mortality. Anastomotic leakage after low anterior resection occurred in 3 of 50 patients (6%; 95% CI, 1-17%). The pathologic complete response rate was 16% (95% CI, 9-26%). Pathologic Stages T2 or less occurred in 51%. CONCLUSION: Preoperative radiotherapy with continuous infusion 5-FU for locally advanced rectal cancer is a safe regimen, with a significant downstaging effect. It does not seem to lead to a significant increase in serious surgical complications. PMID- 11429216 TI - The effectiveness of an immobilization device in conformal radiotherapy for lung tumor: reduction of respiratory tumor movement and evaluation of the daily setup accuracy. AB - PURPOSE: To evaluate the daily setup accuracy and the reduction of respiratory tumor movement using a body frame in conformal therapy for solitary lung tumor. METHODS AND MATERIALS: Eighteen patients with a solitary lung tumor underwent conformal therapy using a body frame. The body shell of the frame was shaped to the patient's body contour. The respiratory tumor movement was estimated using fluoroscopy, and if it was greater than 5 mm, pressure was applied to the patient's abdomen with the goal of minimizing tumor movement. CT images were then obtained, and a treatment planning was made. A total dose of 40 or 48 Gy was delivered in 4 fractions. Portal films were obtained at each treatment, and the field displacements between them and the simulation films were measured for daily setup errors. The patients were repositioned if the setup error was greater than 3 mm. Correlations were analyzed between patient characteristics and the tumor movement, or the tumor movement reduction and the daily setup errors. RESULTS: Respiratory tumor movement ranged from 0 to 20 mm (mean 7.7 mm). The abdominal press reduced the tumor movement significantly from a range of 8 to 20 mm to a range of 2 to 11 mm (p = 0.0002). Daily setup errors were within 5 mm in 90%, 100%, and 93% of all verifications in left-right, anterior-posterior, and cranio caudal directions, respectively. Patient repositioning was performed in 25% of all treatments. No significant correlation was detected between patient characteristics and tumor movement, tumor movement reduction, and the daily setup errors. CONCLUSIONS: The abdominal press was successful in reducing the respiratory tumor movement. Daily setup accuracy using the body frame was acceptable. Verification should be performed at each treatment in hypofractionated conformal therapy. PMID- 11429217 TI - Predicting the risk of symptomatic radiation-induced lung injury using both the physical and biologic parameters V(30) and transforming growth factor beta. AB - PURPOSE: To correlate the volume of lung irradiated with changes in plasma levels of the fibrogenic cytokine transforming growth factor beta (TGFbeta) during radiotherapy (RT), such that this information might be used to predict the development of symptomatic radiation-induced lung injury (SRILI). METHODS AND MATERIALS: The records of all patients with lung cancer treated with RT with curative intent from 1991 to 1997 on a series of prospective normal tissue injury studies were reviewed. A total of 103 patients were identified who met the following inclusion criteria: (1) newly diagnosed lung cancer of any histology treated with RT +/- chemotherapy with curative intent; (2) no evidence of distant metastases or malignant pleural effusion; (3) no thoracic surgery after lung RT; (4) no endobronchial brachytherapy; (5) follow-up time more than 6 months; (6) plasma TGFbeta1 measurements obtained before and at the end of RT. The concentration of plasma TGFbeta1 was measured by an enzyme-linked immunosorbent assay. Seventy-eight of the 103 patients were treated with computed tomography based 3-dimensional planning and had dose-volume histogram data available. The endpoint of the study was the development of SRILI (modified NCI [National Cancer Institute] common toxicity criteria). RESULTS: The 1-year and 2-year actuarial incidence of SRILI for all 103 patients was 17% and 21%, respectively. In those patients whose TGFbeta level at the end of RT was higher than the pre-RT baseline, SRILI occurred more frequently (2-year incidence = 39%) than in patients whose TGFbeta1 level at the end of RT was less than the baseline value (2-year incidence = 11%, p = 0.007). On multivariate analysis, a persistent elevation of plasma TGFbeta1 above the baseline concentration at the end of RT was an independent risk factor for the occurrence of SRILI (p = 0.004). The subgroup of 78 patients treated with 3-dimensional conformal radiotherapy, who consequently had dose-volume histogram data, were divided into groups according to their TGFbeta1 kinetics and whether their V(30) level was above or below the median of 30%. Group I (n = 29), with both a TGFbeta1 level at the end of RT that was below the pre-RT baseline and V(30) < 30%; Group II (n = 35), with a TGFbeta1 level at the end of irradiation that was below the baseline but a V(30) > or = 30% or with a TGFbeta1 level at the end of RT that was above the pre-RT baseline but V(30) < 30%; Group III (n = 14), with both a TGFbeta1 level at the end of RT that was above the baseline and V(30) > or = 30%. A significant difference was found in the incidence of SRILI among these three groups (6.9%, 22.8%, 42.9%, respectively, p = 0.02). CONCLUSIONS: (1) An elevated plasma TGFbeta1 level at the end of RT is an independent risk factor for SRILI; (2) The combination of plasma TGFbeta1 level and V(30) appears to facilitate stratification of patients into low, intermediate, and high risk groups. Thus, combining both physical and biologic risk factors may allow for better identification of patients at risk for the development of symptomatic radiation-induced lung injury. PMID- 11429218 TI - Radiotherapy of testicular intraepithelial neoplasia (TIN): a novel treatment regimen for a rare disease. AB - PURPOSE: Testicular intraepithelial neoplasia (TIN) is a consistent precursor of most invasive germ cell tumors, currently treated by radiotherapy with 20 Gy, which destroys TIN but preserves Leydig cells. Nevertheless, analysis has shown dose-dependent dysfunction even with low therapeutic doses of 20 Gy in some cases. Therefore, we tested a dose reduction regimen by delivering smaller fractional doses to enhance the tolerance of Leydig cells. METHODS AND MATERIALS: Between 1993 and 1999, 9 patients were treated for TIN in a prospective multicenter trial. A total dose of 13 Gy was administered in 10 fractions of 1.3 Gy. Hormonal levels of follicle-stimulating hormone, luteinizing hormone, and testosterone were assayed serially. RESULTS: During a median follow-up time of 36 months, no patient showed evidence of local disease. A first postradiation biopsy was obtained 3-12 months after radiotherapy; 5 patients underwent a second biopsy 2-3 years after treatment. All biopsies showed a Sertoli cell-only pattern. Follicle-stimulating hormone levels continued to increase 1 year after radiotherapy, signaling eradicated spermiogenesis. Luteinizing hormone and testosterone remained within the normal range 2 years after radiotherapy. CONCLUSIONS: In the treatment of TIN, there seems to be a dose reduction potential to 13 Gy by lowering single fractional doses, which enhances the therapeutic ratio in favor of the Leydig cells. PMID- 11429219 TI - MR-spectroscopy guided target delineation for high-grade gliomas. AB - PURPOSE: Functional/metabolic information provided by MR-spectroscopy (MRSI) suggests MRI may not be a reliable indicator of active and microscopic disease in malignant brain tumors. We assessed the impact MRSI might have on the target volumes used for radiation therapy treatment planning for high-grade gliomas. METHODS AND MATERIALS: Thirty-four patients (22 Grade III; 12 Grade IV astrocytomas) were evaluated; each had undergone MRI and MRSI studies before surgery. MRI data sets were contoured for T1 region of contrast enhancement (T1), region of necrosis, and T2 region of hyperintensity (T2). The three-dimensional MRSI peak parameters for choline (Cho) and N-acetylaspartate (NAA), acquired by a multivoxel technique, were categorized based on an abnormality index (AI), a quantitative assessment of tissue metabolite levels. The AI data were aligned to the MRI and displayed as three-dimensional contours. AI vs. T conjoint and disjoint volumes were compared. RESULTS: For both grades, although T2 estimated the region at risk of microscopic disease as being as much as 50% greater than by MRSI, metabolically active tumor still extended outside the T2 region in 88% of patients by as many as 28 mm. In addition, T1 suggested a lesser volume and different location of active disease compared to MRSI. CONCLUSION: The use of MRSI to define target volumes for RT treatment planning would increase, and change the location of, the volume receiving a boost dose as well as reduce the volume receiving a standard dose. Incorporation of MRSI into the treatment planning process may have the potential to improve control while reducing complications. PMID- 11429220 TI - Radiosurgery in the management of pediatric brain tumors. AB - OBJECTIVE: To describe the outcome of pediatric brain tumor patients following stereotactic radiosurgery (SRS), and factors associated with progression-free survival. METHODS: We reviewed the outcome of 90 children treated with SRS for recurrent (n = 62) or residual (n = 28) brain tumors over a 10-year period. Median follow-up from SRS was 24 months for all patients and 55.5 months for the 34 patients currently alive. RESULTS: The median progression-free survival (PFS) for all patients was 13 months. Median PFS according to tumor histology was medulloblastoma = 11 months, ependymoma = 8.5 months, glioblastoma and anaplastic astrocytoma = 12 months. Median PFS in patients treated to a single lesion was 15.4 months. No patient undergoing SRS to more than 1 lesion survived disease free beyond 2 years. After adjusting for histology and other clinical factors, SRS for tumor recurrence (RR = 2.49) and the presence of > 1 lesion (RR = 2.3) were associated with a significantly increased rate of progression (p < 0.05). Three-year actuarial local control (LC) was as follows: medulloblastoma = 57%, ependymoma = 29%, anaplastic astrocytoma/glioblastoma = 60%, other histologies = 56%. Nineteen patients with radionecrosis and progressive neurologic symptoms underwent reoperation after an interval of 0.6-62 months following SRS. Pathology revealed necrosis with no evidence of tumor in 9 of these cases. CONCLUSION: SRS can be given safely to selected children with brain tumors. SRS appears to reduce the proportion of first failures occurring locally and is associated with better outcome when given as a part of initial management. Some patients with unresectable relapsed disease can be salvaged with SRS. SRS to multiple lesions does not appear to be curative. Serious neurologic symptoms requiring reoperation is infrequently caused by radionecrosis alone. PMID- 11429221 TI - Acute urinary toxicity following transperineal prostate brachytherapy using a modified Quimby loading method. AB - PURPOSE: To examine the acute urinary toxicity following transperineal prostate implant using a modified Quimby loading method with regard to time course, severity, and factors that may be associated with a higher incidence of morbidity. METHODS AND MATERIALS: One hundred thirty-nine patients with prostate adenocarcinoma treated with brachytherapy from 1997 through 1999 had follow-up records available for review. Patients considered for definitive brachytherapy alone included those with prostate specific antigen (PSA) < or = 6, Gleason score (GS) < or = 6, clinical stage < T2b, and prostate volumes generally less than 40 cc. Patients with larger prostate volumes were given neoadjuvant antiandrogen therapy. Those with GS > 6, PSA > 6, or Stage > T2a were treated with external beam radiation therapy followed by brachytherapy boost. Sources were loaded according to a modified Quimby method. At each follow-up, toxicity was graded based on a modified RTOG urinary toxicity scale. RESULTS: Acute urinary toxicity occurred in 88%. Grade I toxicity was reported in 23%, grade II in 45%, and grade III in 20%, with 14% requiring prolonged (greater than 1 week) intermittent or indwelling catheterization. Overall median duration of symptoms was 12 months. There was no difference in duration of symptoms between patients treated with I 125 or Pd-103 sources (p = 0.71). After adjusting for GS and PSA, multivariate logistic regression analysis showed higher incidence of grade 3 toxicity in patients with larger prostate volumes (p = 0.002), and those with more seeds implanted (p < 0.001). Higher incidence of prolonged catheterization was found in patients receiving brachytherapy alone (p = 0.01), with larger prostate volumes (p = 0.01), and those with more seeds implanted (p < 0.001). CONCLUSION: Interstitial brachytherapy for prostate cancer leads to a high incidence of acute urinary toxicity, most of which is mild to moderate in severity. A prolonged need for catheterization can occur in some patients. Patients receiving brachytherapy alone, those with prostate volumes greater than 30 cc, and those implanted with a greater number of seeds have the highest incidence of significant toxicity. PMID- 11429222 TI - To move or not to move: measurements of prostate motion by urethrography using MRI. AB - PURPOSE: Urethrography is commonly used to aid in definition of the prostate apex during CT simulation for prostate cancer. If the position of the prostate were altered by the urethrogram itself, then systematic error could be introduced into the patient's treatment. Sagittal MRI scans were acquired immediately before and after a localization urethrogram to determine the extent of displacement. METHODS AND MATERIALS: Thirteen patients underwent sagittal T2-weighted fast spin echo MRI scans. Patients were scanned supine in an alpha cradle cast in the treatment position. The prostate was contoured by 3 different observers to determine the apex location on the central sagittal MRI section and the center of mass relative to an immobile bony landmark. Statistical multivariate analysis was performed to establish if there was a net displacement of the prostate (systematic error), and to determine the margin required to cover the random prostate position within a 95% confidence interval. RESULTS: There was no significant systematic motion of either the prostate nor its apex in either the anterior-posterior or superior inferior directions. The average motion of the prostate center of mass was 0.04 +/- 0.40 cm (1 SD) and 0.01 +/- 0.33 cm in the anterior-posterior and superior inferior direction, respectively. The corresponding figures for location of the apex were 0.05 +/- 0.30 cm and 0.01 +/- 0.33 cm, respectively. The statistical analysis revealed that a margin of 2 mm is sufficient to cover any random motion of the prostate that could occur as a result of the urethrogram 95% of the time. CONCLUSION: Urethrography during CT simulation for prostate cancer does not cause significant prostate displacement or systematic error in planning and delivering external-beam radiation. PMID- 11429223 TI - Treatment of early and moderately advanced vocal cord carcinoma with 6-MV X-rays. AB - PURPOSE: Whereas there are many reports regarding treatment of early vocal cord cancer with cobalt 60 or 2-4-MV X-rays, there are still few reports on the results of treatment with 6-MV X-rays. Theoretically, 6-MV X-rays result in greater underdosage of tumor at the air-tissue interface and at the anterior commissure. This paper analyzes the results of irradiation of early and moderately advanced squamous cell carcinoma of the true vocal cord treated exclusively with 6-MV X-rays in a community hospital. The literature pertinent to the issue is reviewed. METHODS AND MATERIALS: Eighty-three patients with Tis, T1, T2, or T3 squamous cell carcinoma of the true vocal cord were treated with curative intent at Bethesda Memorial Hospital in Boynton Beach, Florida between April 1986 and April 1998. The dose schedules most commonly used were 63 Gy in 28 fractions (2.25 Gy per fraction once a day) for T1 tumors or 74.40 Gy in 62 fractions (1.2 Gy per fraction twice a day) for T2 and T3 tumors. All patients have minimum 2-year follow-up; 63 (76%) have 5-year minimum follow-up. RESULTS: Local control was achieved in 6 of 6 Tis, 53 of 54 (98%) T1, 8 of 8 T2, and 6 of 6 T3 lesions. No complications were encountered. CONCLUSIONS: A recent literature review indicates that the treatment of early vocal cord cancer with 6-MV X-rays remains controversial. The dose schedules used in the present paper produced a high rate of local control, a finding that is consistent with reports of other investigators who used dose schedules similar to those used in the present series. However, several other investigators have reported significantly lower rates of local control for T1 or T2 glottic cancer treated with 6-MV X-rays when compared to results obtained with cobalt 60 or 4 MV at their own institution. The latter institutions used lower total doses and/or lower dose per fraction than those institutions reporting high rates of local control with 6 MV. Data from the literature, as well as our own data, are consistent with the following hypotheses: (1) the lower rates of local control reported by several institutions when using 6 MV compared with cobalt or 2-4 MV, using the same radiation dose schedules for each beam energy, indicate that underdosage of mucosal surfaces in the laryngeal air cavity may be a clinically important phenomenon, and (2) time dose factors, although certainly important for lower energy beams, may be even more important when using 6 MV. PMID- 11429224 TI - Implications of quantitative tumor and nodal regression rates for nasopharyngeal carcinomas after 45 Gy of radiotherapy. AB - PURPOSE: To quantitatively investigate the clinical implications of tumor regression rate (TRR-45) and nodal regression rate (NRR-45) of nasopharyngeal carcinomas (NPC) after receiving 45 Gy of radiotherapy (RT). The values, predictive values, and associated factors of TRR-45 and NRR-45 in NPC are analyzed. METHODS AND MATERIALS: One hundred one patients with newly diagnosed NPC and who were curatively treated by RT alone were included in the study. Tumor volume and nodal volume before treatment and after 45 Gy were obtained from computed tomographic (CT) scans performed at those times and calculated with the assistance of a computer-based imaging analyzing system. TRR-45 (NRR-45) was defined as the ratio of reduced tumor (nodal) volume after 45 Gy to the initial tumor (nodal) volume. TRR-45 (NRR-45) values were stratified into three groups of slow (below 50%), moderate (between 50% and 75%), and rapid (above 75%) change. After conventional RT with 45 Gy, conformal RT for primary tumors was boosted to 70.2-72 Gy for T1-2 tumors, and 75.6-81 Gy for T3-T4 tumors. RT for residual neck masses was boosted by electron beam to 61-75 Gy. RESULTS: The mean value of TRR 45 for all patients was lower than that of NRR-45 for the 78 patients with metastatic neck nodes (70% +/- 4.8% vs. 81% +/- 5%, p = 0.003). The 3-year actuarial neck control rate was better than the primary tumor control rate with statistical significance (98% vs. 85%, p = 0.009). No significant statistical differences concerning local control probability, nodal control probability, or survival rate were found among patients with slow, moderate, or rapid TRR-45 or NRR-45. T-stage was the only significant prognostic factor for locoregional control after multivariate analysis. Tumor volume and T-stage were found to have a statistically significant negative correlation with TRR-45. No associated factor was found to be significantly correlated with NRR-45. CONCLUSION: Slow regression rates of the primary tumor or neck nodes in NPC after receiving 45 Gy of irradiation do not mean ultimately poor radiocurability, but may merely imply slow clearance of the cells damaged during irradiation. The different radiobiological behaviors of the regression rates during treatment, ultimate control probabilities, or associated factors for regression rates of NPC between primary tumors and neck nodes need to be further investigated. PMID- 11429225 TI - Mantle irradiation alone for pathologic stage I and II Hodgkin's disease: long term follow-up and patterns of failure. AB - PURPOSE: We performed a retrospective study to determine the long-term outcome, patterns of failure, and prognostic factors for patients with pathologic Stage I or II Hodgkin's disease (HD) who were treated with mantle irradiation alone. METHODS AND MATERIALS: The medical records of 145 patients with pathologic Stage I or II supradiaphragmatic Hodgkin's disease treated with mantle irradiation alone between June 1967 and June 1991 were reviewed. Patterns of failure, overall survival (OS) rate, and progression-free survival (PFS) rate were determined. Univariate and multivariate analyses were performed to identify adverse prognostic factors for OS and PFS. The number of adverse prognostic factors per patient was counted, and a prognostic score was assigned to each patient. The log rank test was used to compare the OS or PFS rates among patients with prognostic scores 0, 1, and 2. RESULTS: The median patient age was 27 years (range 10-66), with almost even male to female distribution. Every patient had splenectomy and negative laparotomy (LAP). Fifty-one patients had Stage I disease (IA-49, IB-2) and 94 Stage II (IIA-89, IIB-5). The histologic subtypes were nodular sclerosing in 110, mixed cellularity in 28, lymphocyte predominance in 5, lymphocyte depleted in 1, and unclassified in 1. Twelve patients with Stage II disease had >/= 3 sites of nodal involvement. Fifty-four patients had a prognostic score of 0, 70 of 1, and 21 of 2. The median follow-up time for the 109 surviving patients was 146 months (range 25-381). The 10- and 20-year actuarial OS rates for the whole group were 87.6% and 65.3%, respectively. The corresponding actuarial PFS rates were 75.3% and 74.2%, respectively. Thirty-six patients (9 Stage I, 27 Stage II) had relapses in a total of 41 sites. Failures by histology were 29 patients with nodular sclerosing, 6 with mixed cellularity, and 1 with lymphocyte predominance. Failures by sites were: trans-diaphragmatic, 22 (para-aortic nodes, 15; as the only site of progression in 12; visceral, 7; as the only site of progression in 5); within radiation field, 8; marginal miss, 8 (as the only site of failure in 2); and unknown, 3. The majority of the failures occurred within 5 years of diagnosis. Long-term side effects of radiation included cardiac complications in 30 patients, with 10- and 20-year actuarial cardiac complication rates of 12.6% and 35.1%, respectively; secondary solid tumors in 14, with 10- and 20-year actuarial rates of 2.3% and 25.7%, respectively; leukemia in 4; non- Hodgkin's lymphoma in 4, with the 10- and 20-year actuarial rates for leukemia and non-Hodgkin's lymphoma of 4.0% and 13.9%; and hypothyroidism in 38. Four adverse prognostic factors were identified for PFS: age > or = 40 years, > or = 3 sites of involvement, male sex, and constitutional symptoms. The prognostic score correlated with patients' outcome as indicated by PFS and OS rates. Patients with a prognostic score of 0 did significantly better than those with a score of 1 or 2. CONCLUSION: In this select group of patients with pathologic Stage I and II Hodgkin's disease treated with mantle irradiation alone, the OS and PFS rates at 10 and 20 years were comparable to those reported in the literature. The major pattern of disease progression was relapse below the diaphragm, therefore close surveillance of the abdomen is warranted. The prognostic score used in our series may predict the patient's outcome, and might be worth testing in a prospective trial. In our series, patients with a prognostic score of 0 had excellent long term survival, indicating adequate treatment with mantle irradiation alone. Late complications of the treatment pose a significant threat for the patient's survival with long-term follow-up. PMID- 11429226 TI - Costs and effectiveness of staging and treatment options in early-stage Hodgkin's disease. AB - PURPOSE: Using a cost-effectiveness analysis, to weigh the costs and benefits of the different staging and treatment options in early-stage Hodgkin's disease. METHODS: We constructed a decision-analytic model for a hypothetical cohort of 25 year-old patients with early-stage Hodgkin's disease. Markov models were used to simulate the lifetime costs and prognosis of each staging and treatment strategy. Baseline probabilities and cost estimates were derived from published studies and bills of relevant patient cohorts. RESULTS: Among the six management strategies considered, the incremental cost-effectiveness ratio of laparotomy and tailored treatment compared with mantle and para-aortic-splenic radiation therapy in all clinical stage I-II patients was $24,100/quality-adjusted life year, while that of the strategy of combined modality therapy in all clinical stage I-II patients compared with laparotomy was $61,700/quality-adjusted life year. All the remaining strategies were dominated by one of these three strategies. Sensitivity analysis showed that the cost-effectiveness ratios were driven predominantly by the effectiveness rather than the cost of each strategy. In particular, the analysis was heavily influenced by the utility of the post-laparotomy health state. CONCLUSIONS: In considering the various alternative management strategies in early-stage Hodgkin's disease, even very small gains in effectiveness were enough to justify the additional costs of more expensive treatment options. PMID- 11429227 TI - Mammographically detected ductal carcinoma in situ of the breast treated with breast-conserving surgery and definitive breast irradiation: long-term outcome and prognostic significance of patient age and margin status. AB - PURPOSE: This study was performed to determine the long-term outcome for women with mammographically detected ductal carcinoma in situ (DCIS; intraductal carcinoma) of the breast treated with breast-conserving surgery followed by definitive breast irradiation. METHODS AND MATERIALS: An analysis was performed of 422 mammographically detected intraductal breast carcinomas in 418 women from 11 institutions in North America and Europe. All patients were treated with breast-conserving surgery followed by definitive breast irradiation. The median follow-up time was 9.4 years (mean, 9.4 years; range, 0.1-19.8 years). RESULTS: The 15-year overall survival rate was 92%, and the 15-year cause-specific survival rate was 98%. The 15-year rate of freedom from distant metastases was 94%. There were 48 local failures in the treated breast, and the 15-year rate of any local failure was 16%. The median time to local failure was 5.0 years (mean, 5.7 years; range, 1.0-15.2 years). Patient age at the time of treatment and final pathology margin status from the primary tumor excision were both significantly associated with local failure. The 10-year rate of local failure was 31% for patient age < or = 39 years, 13% for age 40-49 years, 8% for age 50-59 years, and 6% for age > or = 60 years (p = 0.0001). The 10-year rate of local failure was 24% when the margins of resection were positive, 9% when the margins of resection were negative, 7% when the margins of resection were close, and 12% when the margins of resection were unknown (p = 0.030). Patient age < or = 39 years and positive margins of resection were both independently associated with an increased risk of local failure (p = 0.0006 and p = 0.023, respectively) in the multivariable Cox regression model. CONCLUSIONS: The 15-year results from the present study demonstrated high rates of overall survival, cause-specific survival, and freedom from distant metastases following the treatment of mammographically detected ductal carcinoma in situ of the breast using breast conserving surgery and definitive breast irradiation. Younger age and positive margins of resection were both independently associated with an increased risk of local failure. The 15-year results in the present study serve as an important benchmark for comparison with other treatment modalities. These results support the use of breast-conserving surgery and definitive breast irradiation for the treatment of appropriately selected patients with mammographically detected ductal carcinoma in situ of the breast. PMID- 11429228 TI - The use of high-dose-rate brachytherapy alone after lumpectomy in patients with early-stage breast cancer treated with breast-conserving therapy. AB - PURPOSE: We present the preliminary results of our in-house protocol using outpatient high-dose-rate (HDR) brachytherapy as the sole radiation modality following lumpectomy in patients with early-stage breast cancer. METHODS AND MATERIALS: Thirty-seven patients with 38 Stage I-II breast cancers received radiation to the lumpectomy cavity alone using an HDR interstitial implant with (192)Ir. A minimum dose of 32 Gy was delivered on an outpatient basis in 8 fractions of 4 Gy to the lumpectomy cavity plus a 1- to 2-cm margin over consecutive 4 days. RESULTS: Median follow-up is 31 months. There has been one ipsilateral breast recurrence for a crude failure rate of 2.6% and no regional or distant failures. Wound healing was not impaired in patients undergoing an open cavity implant. Three minor breast infections occurred, and all resolved with oral antibiotics. The cosmetic outcome was good to excellent in all patients. CONCLUSION: In selected patients with early-stage breast cancer, treatment of the lumpectomy cavity alone with outpatient HDR brachytherapy is both technically feasible and well tolerated. Early results are encouraging, however, longer follow-up is necessary before equivalence to standard whole-breast irradiation can be established and to determine the most optimal radiation therapy technique to be employed. PMID- 11429229 TI - Extent and kinetics of recovery of occult spinal cord injury. AB - PURPOSE: To obtain clinically useful quantitative data on the extent and kinetics of recovery of occult radiation injury in primate spinal cord, after a commonly administered elective radiation dose of 44 Gy, given in about 2 Gy per fraction. METHODS AND MATERIALS: A group of 56 rhesus monkeys was assigned to receive two radiation courses to the cervical and upper thoracic spinal cord, given in 2.2 Gy per fraction. The dose of the initial course was 44 Gy in all monkeys. Reirradiation dose was 57.2 Gy, given after 1-year (n = 16) or 2-year (n = 20) intervals, or 66 Gy, given after 2-year (n = 4) or 3-year (n = 14) intervals. Two animals developed intramedullary tumors before reirradiation and, therefore, did not receive a second course. The study endpoint was myeloparesis, manifesting predominantly as lower extremity weakness and decrease in balance, occurring within 2.5 years after reirradiation, complemented by histologic examination of the spinal cord. The data obtained were analyzed along with data from a previous study addressing single-course tolerance, and data from a preliminary study of reirradiation tolerance. RESULTS: Only 4 of 45 monkeys completing the required observation period (2-2.5 years after reirradiation, 3-5.5 years total) developed myeloparesis. The data revealed a substantial recovery of occult injury induced by 44 Gy within the first year, and suggested additional recovery between 1 and 3 years. Fitting the data with a model, assuming that all (single course and reirradiation) dose-response curves were parallel, yielded recovery estimates of 33.6 Gy (76%), 37.6 Gy (85%), and 44.6 Gy (101%) of the initial dose, after 1, 2, and 3 years, respectively, at the 5% incidence (D(5)) level. The most conservative estimate, using a model in which it was assumed that there was no recovery between 1 and 3 years following initial irradiation and that the combined reirradiation curve was not necessarily parallel to the single-course curve, still showed an overall recovery equivalent to 26.8 Gy (61%). The spinal cords of symptomatic monkeys consistently revealed a mixture of white matter necrosis and vascular injury, but the majority of spinal cords of asymptomatic animals did not exhibit overt lesions detectable by light microscopy. CONCLUSION: Combined analysis with the data of the previous studies yielded firm evidence that the spinal cord has a large capacity to recover from occult radiation injury induced by a commonly prescribed elective dose. This finding strengthens the rationale for selective use of radiotherapy to treat second primary tumors arising in previously irradiated tissues or late recurrences. However, some caution should be exercised in applying quantitative experimental data, because the length of follow-up in these experiments was limited to 2-2.5 years after reirradiation, whereas human myelopathy cases occasionally occur after longer latency. Because there is a large variation in long-term recovery among tissues, the tolerance of other tissues at risk should also be taken into account in prescribing therapy. PMID- 11429230 TI - Is alpha/beta for prostate tumors really low? AB - PURPOSE: Brenner and Hall's 1999 paper estimating an alpha/beta value of 1.5 Gy for prostate tumors has stimulated much interest in the question of whether this ratio (of intrinsic radiosensitivity to repair capacity) is much lower in prostate tumors than in other types of tumors that proliferate faster. The implications for possibly treating prostatic cancer using fewer and larger fractions are important. In this paper we review updated clinical data and present somewhat different calculations to estimate alpha/beta. METHODS AND MATERIALS: Seventeen clinical papers published from 1995 to 2000 were reviewed to obtain estimates of biochemical control from radiotherapy alone using external beam, I-125 implants, or Pd-103 implants. The focus was on intermediate risk patients. Three methods of estimating alpha/beta were employed. First, a simple two-step graphical comparison of isoeffective doses from external beam and implant modalities was made, to see which value of alpha/beta predicted the observed identity of biologic effect. Second, the same data were subjected to Direct Analysis (maximum likelihood estimation), from which an estimate of alpha/beta and also of the T(12) of repair of sublethal damage in the tumors (both with confidence intervals) were obtained. Third, preliminary clinical data comparing two different sizes of high-dose boost doses were analyzed in which significantly different bNED was observed at 2 years. RESULTS: The second method gave the definitive result of alpha/beta = 1.49 Gy (95% CI 1.25-1.76) and T(12) = 1.90 h (95% CI 1.42-2.86 h). The first method gave a range from 1.4 to 1.9 Gy and showed that if mean or median dose were used instead of prescribed dose, the estimate of alpha/beta would be substantially below 1 Gy. The third method, although based on early follow-up, was consistent with low values of alpha/beta in the region of 2 Gy or below. The estimate for T(12) is the first value reported for prostate tumors in situ. CONCLUSIONS: All the estimates point toward low values of alpha/beta, at least as low as the estimates of Brenner and Hall, and possibly lower than the expected values of about 3 Gy for late complications. Hypofractionation trials for intermediate-risk prostatic cancer appear to be indicated. PMID- 11429231 TI - Decreased adventitial neovascularization after intracoronary irradiation in swine: a time course study. AB - BACKGROUND: Intracoronary radiation (IR) suppresses the formation of neointima after arterial injury in swine, through mechanisms incompletely understood. Neointimal development appears related to expansion of adventitial microvessels; we therefore examined the hypothesis that IR inhibits neointima formation through an anti-angiogenic effect. METHODS AND RESULTS: Juvenile swine were treated with either 0 or 15 Gy (192)Ir (gamma-source) and euthanized 3, 7, or 14 days later or treated with 18 Gy (90)Y (beta-source) and euthanized after 14 days. Adventitial area (AA), intimal area (IA), IA corrected for medial fracture length, and adventitial vessel area were assessed in both injured and uninjured segments by computer-aided histomorphometry on Verhoeff-Von Giesson stained sections. Adventitial vessel count (AVC) was enumerated visually on hematoxylin and eosin stained sections and confirmed by anti-factor VIII-associated antigen immunostaining for endothelial cells. AA and IA were reduced in injured arteries subjected to IR as compared to controls. The AVC was significantly lower in injured irradiated arterial segments as well as all uninjured segments as compared with injured control segments. In the injured and irradiated arteries, the AVC remained unchanged at 3, 7, and 14 days. The injured segments of arteries treated with IR demonstrated a significantly lower adventitial microvessel density (AVC/AA) as compared to the injured control segments. Comparison of gamma and beta-irradiation at 14 days did not show any differences for vessel parameters and measurements of adventitial microvessels. IA and AVC were correlated positively (R(2) = 0.63, alpha = 0.79, p < 0.01). CONCLUSION: IR induced an early and sustained anti-angiogenic effect between 3 and 14 days. The relation between IA and AVC may indicate an antiproliferative effect associated with an anti-angiogenic effect independent of the type of radiation. CONDENSED ABSTRACT: Intracoronary radiation suppresses neointima formation after arterial injury in swine, through mechanisms and with consequences that are not fully known. Reduction of angiogenesis may inhibit restenosis. In the present study, intimal area and adventitial area were reduced in the intracoronary radiation groups 3-14 days after arterial injury as compared to their respective controls, with a parallel reduction in the adventitial vessel count and adventitial vessel density. Intimal area and adventitial vessel count were correlated positively. Neointima reduction after intracoronary radiation may depend not only on an antiproliferative effect but also on an anti-angiogenic effect. PMID- 11429232 TI - The role of protein kinase B (PKB) in modulating heat sensitivity in a human breast cancer cell line. AB - PURPOSE: Protein kinase B (PKB) is a critical mediator of phosphoinositide 3 kinase-dependent survival signals in mammalian cells. Its activity is induced after heat shock, and is inhibited in cells undergoing apoptosis. We hypothesized that PKB may be an important modulator for heat-induced apoptosis in human cancer cells. METHODS AND MATERIALS: MCF-7 cells were transfected using four different plasmids, encoding a kinase-dead mutant PKB-AAA, a constitutively activated mutant PKB-DD, wild-type PKB, and the neomycin-resistant selection gene. These stable transfectants were subjected to heat shock, and assessed for PKB phosphorylation, PKB activity, and likelihood of undergoing apoptosis. RESULTS: After heating to 45 degrees C x 30 mins, 25% of MCF-7/neo transfectants underwent apoptosis, which increased to 38% in the presence of wortmannin (WT), an inhibitor of phosphoinositide 3-kinase. In contrast, 23% of the constitutively activated MCF-7/DD transfectants underwent apoptosis, minimally affected by WT. Heat-induced apoptosis occurred in 34% of the kinase-dead MCF-7/AAA transfectants, which increased further to 58% with the addition of WT. This in turn was associated with a two-fold reduction in clonogenic survival compared to the MCF-7/neo transfectants. CONCLUSION: Heat shock activation of PKB in human MCF-7 cells appears to be a significant modulator of heat-induced apoptosis and survival. Further understanding of this important pathway may offer potential in developing novel strategies in cancer therapy. PMID- 11429233 TI - Heterogeneity in 2-deoxy-D-glucose-induced modifications in energetics and radiation responses of human tumor cell lines. AB - PURPOSE: The glucose analog and glycolytic inhibitor, 2-deoxy-D-glucose (2-DG), has been shown to differentially enhance the radiation damage in tumor cells by inhibiting the postirradiation repair processes. The present study was undertaken to examine the relationship between 2-DG-induced modification of energy metabolism and cellular radioresponses and to identify the most relevant parameter(s) for predicting the tumor response to the combined treatment of radiation + 2-DG. METHODS AND MATERIALS: Six human tumor cell lines (glioma: BMG 1 and U-87, squamous cell carcinoma: 4451 and 4197, and melanoma: MeWo and Be-11) were investigated. Cells were exposed to 2 Gy of Co-60 gamma-rays or 250 kVP X rays and maintained under liquid-holding conditions 2-4 h to facilitate repair. 2 DG (5 mM, equimolar with glucose) that was added at the time of irradiation was present during the liquid holding. Glucose utilization, lactate production (enzymatic assays), and adenine nucleotides (high performance liquid chromatography and capillary isotachophoresis) were investigated as parameters of energy metabolism. Induction and repair of DNA damage (comet assay), cytogenetic damage (micronuclei formation), and cell death (macrocolony assay) were analyzed as parameters of radiation response. RESULTS: The glucose consumption and lactate production of glioma cell lines (BMG-1 and U-87) were nearly 2-fold higher than the squamous carcinoma cell lines (4197 and 4451). The ATP content varied from 3.0 to 6.5 femto moles/cell among these lines, whereas the energy charge (0.86 0.90) did not show much variation. Presence of 2-DG inhibited the rate of glucose usage and glycolysis by 30-40% in glioma cell lines and by 15-20% in squamous carcinoma lines, while ATP levels reduced by nearly 40% in all the four cell lines. ATP:ADP ratios decreased to a greater extent ( approximately 40%) in glioma cells than in squamous carcinoma 4451 and MeWo cells; in contrast, presence of 2-DG reduced ADP:AMP ratios by 3-fold in the squamous carcinoma 4451, whereas an increase was noted in the glioma cell line BMG-1. 2-DG significantly reduced the initial rates of DNA repair in all cells, resulting in an excess residual damage after 2 h of repair in BMG-1, U-87, and 4451 cell lines, whereas no significant differences could be observed in the other cell lines. Recovery from potentially lethal damage was also significantly inhibited in BMG-1 cells. 2 DG increased the radiation-induced micronuclei formation in the melanoma line (MeWo) by nearly 60%, while a moderate (25-40%) increase was observed in the glioma cell lines (BMG-1 and U-87). Presence of 2-DG during liquid holding (4 h) enhanced the radiation-induced cell death by nearly 40% in both the glioma cell lines, while significant effects were not observed in others. CONCLUSIONS: The modifications in energetics and radiation responses by 2-DG vary considerably among different human tumor cell lines, and the relationships between energy metabolism and various radiobiologic parameters are complex in nature. The 2-DG induced modification of radiation response does not strictly correlate with changes in the levels of ATP. However, a significant enhancement of the radiation damage by 2-DG was observed in cells with high rates of glucose usage and glycolysis, which appear to be the two most important factors determining the tumor response to the combined treatment of 2-DG + radiation therapy. PMID- 11429234 TI - Radiosensitization effect by combination with paclitaxel in vivo, including the effect on intratumor quiescent cells. AB - PURPOSE: To evaluate the radiosensitization effect on solid tumors upon combination treatment with paclitaxel (TXL), including the effect on intratumor quiescent (Q) cells. METHODS AND MATERIALS: Mice bearing SCC VII or EL4 solid tumors received 5-bromo-2'-deoxyuridine (BrdU) continuously for 5 days to label all proliferating (P) cells. The mice then received gamma-irradiation with or without tirapazamine (TPZ) at various time points after TXL administration. Another group of mice received a series of test doses of gamma-rays while alive or after tumor clamping to obtain hypoxic fractions (HFs) in the tumors at various time points after TXL administration. Immediately after irradiation, the tumor cells were isolated and incubated with a cytokinesis blocker. The micronucleus (MN) frequency in cells without BrdU labeling (Q cells) was determined using immunofluorescence staining for BrdU. Meanwhile, 6 h after irradiation, the tumor cells were isolated from the solid tumors in another group of mice, and the apoptosis frequency in Q cells was also determined with immunofluorescence staining for BrdU. The MN and apoptosis frequency in total (P + Q) tumor cells were determined from the tumors that were not pretreated with BrdU. For the measurement of the HFs, the MN or apoptosis frequency of Q cells was then used to calculate the surviving fraction of Q cells from the regression line for the relationship between the MN or apoptosis frequency and the surviving fraction of total tumor cells. RESULTS: In both SCC VII and EL4 tumors, maximum values of mitotic index (MI) and apoptosis frequency were observed 9 and 24 h after TXL administration, respectively. However, on the whole, the apoptosis frequency for SCC VII was very low. gamma-Irradiation 9 h after TXL administration induced significant radiosensitization effects on the total cells of both tumors. Irradiation at 60 h had a more significant effect on total cells of EL4 tumor, but no significant effect on total cells of SCC VII tumor. Combined treatment with TXL induced no radiosensitization effect on Q cells in either tumor. The effect on Q cells was observed only after TPZ was administered. The HF of total cells in EL4 tumors decreased significantly 60 h after TXL administration. CONCLUSION: No radiosensitization effect upon combination treatment with TXL is induced in Q tumor cells. However, the effect on P cells is produced by irradiation at the time when the maximum values of MI are induced following TXL administration. In addition, for tumors that are susceptible to apoptosis after TXL administration alone, irradiation at the time of sufficient reoxygenation in tumors after TXL administration produces a greater radioenhancement effect on P cells. PMID- 11429235 TI - Influence of preoperative combined radiochemotherapy on surgical outcome and colonic anastomotic healing: experimental study in the rat. AB - PURPOSE: To study the influence of combined preoperative hyperfractionated irradiation with intraperitoneal 5-fluorouracil (5-FU) on surgical outcome and colonic anastomotic healing in a rat model. METHODS: Male Wistar rats were given 41.6 Gy of preoperative radiotherapy (RT) or sham irradiation, with intraperitoneal 5-FU at low dose (10 mg/kg) or high dose (20 mg/kg). Animals were arranged in 6 groups: RT + low-dose 5-FU (RCT-L), RT + high-dose 5-FU (RCT-H), sham RT + low-dose 5-FU (CT-L), sham RT + high-dose 5-FU (CT-H), RT alone (R), and a control group (sham RT + intraperitoneal saline). Side-to-side colonic anastomoses were constructed from one irradiated and one nonirradiated limb 4 days after radiochemotherapy. Animals were sacrificed 10 days after surgery. RESULTS: Compared to controls, more complications occurred in group RCT-H (50% versus 0%, p = 0.01). Adhesion formation was more intense in groups RCT-H and CT H (p < 0.001 and p = 0.001, respectively). After therapy, white blood cell counts dropped significantly in all irradiated animals (p < 0.01), and platelet counts decreased significantly in group RCT-H (p = 0.01). No significant differences were noticed in anastomotic bursting pressure when the treated groups were compared to each other or to the control group. CONCLUSIONS: Neoadjuvant radiochemotherapy has no adverse effect on the strength of colonic anastomosis in this rat model. However, the combined RT with high-dose 5-FU does increase operative morbidity and adhesion formation. PMID- 11429236 TI - Biology-based combined-modality radiotherapy: workshop report. AB - PURPOSE: The purpose of this workshop summary is to provide an overview of preclinical and clinical data on combined-modality radiotherapy. METHODS AND MATERIALS: The 8th Annual Radiation Workshop at Round Top was held April 13-16, 2000 at the International Festival Institute (Round Top, TX). RESULTS: Presentations by 30 speakers (from Germany, Netherlands, Australia, England, and France along with U.S. participants and M. D. Anderson Cancer Center faculty) formed the framework for discussions on the current status and future perspectives of biology-based combined-modality radiotherapy. CONCLUSION: Cellular and molecular pathways available for radiation modification by chemical and biologic agents are numerous, providing new opportunities for translational research in radiation oncology and for more effective combined-modality treatment of cancer. PMID- 11429237 TI - Dosimetric evaluation of lung tumor immobilization using breath hold at deep inspiration. AB - PURPOSE: To examine the dosimetric benefit of self-gated radiotherapy at deep inspiration breath hold (DIBH) in the treatment of patients with non-small-cell lung cancer (NSCLC). The relative contributions of tumor immobilization at breath hold (BH) and increased lung volume at deep inspiration (DI) in sparing high-dose lung irradiation (> or = 20 Gy) were examined. METHODS AND MATERIALS: Ten consecutive patients undergoing radiotherapy for Stage I-IIIB NSCLC who met the screening criteria were entered on this study. Patients were instructed to BH at DI without the use of external monitors or breath-holding devices (self-gating). Computed tomography (CT) scans of the thorax were performed during free breathing (FB) and DIBH. Fluoroscopy screened for reproducible tumor position throughout DIBH, and determined the maximum superior-inferior (SI) tumor motion during both FB and DIBH. Margins used to define the planning target volume (PTV) from the clinical target volume included 1 cm for setup error and organ motion, plus an additional SI margin for tumor motion, as determined from fluoroscopy. Three conformal treatment plans were then generated for each patient, one from the FB scan with FB PTV margins, a second from the DIBH scan with FB PTV margins, and a third from the DIBH scan with DIBH PTV margins. The percent of total lung volume receiving > or = 20 Gy (using a prescription dose of 70.9 Gy to isocenter) was determined for each plan. RESULTS: Self-gating at DIBH was possible for 8 of the 10 patients; 2 patients were excluded, because they were not able to perform a reproducible DIBH. For these 8 patients, the median BH time was 23 (range, 19-52) s. The mean percent of total lung volume receiving > or = 20 Gy under FB conditions (FB scan with FB PTV margins) was 12.8%. With increased lung volume alone (DIBH scan with FB PTV margins), this was reduced to 11.0%, tending toward a significant decrease in lung irradiation over FB (p = 0.086). With both increased lung volume and tumor immobilization (DIBH scan with DIBH PTV margins), the mean percent lung volume receiving > or = 20 Gy was further reduced to 8.8%, a significant decrease in lung irradiation compared to FB (p = 0.011). Furthermore, at DIBH, the additional benefit provided by tumor immobilization (i.e., using DIBH instead of FB PTV margins) was also significant (p = 0.006). The relative contributions of tumor immobilization and increased lung volume toward reducing the percent total lung volume receiving > or = 20 Gy were patient specific; however, all 8 of the patients analyzed showed a dosimetric benefit with this DIBH technique. CONCLUSION: Compared to FB conditions, at DIBH the mean reduction in percent lung volume receiving > or = 20 Gy was 14.3% with the increase in lung volume alone, 22.1% with tumor immobilization alone, and 32.5% with the combined effect. The dosimetric benefit seen at DIBH was patient specific, and due to both the increased lung volume seen at DI and the PTV margin reduction seen with tumor immobilization. PMID- 11429238 TI - Fraxinus ornus L. AB - The literature on the chemical constituents and the biological activity of Fraxinus ornus bark, leaves and flowers has been reviewed. Chemical studies show the presence of many compounds belonging mainly to the groups of hydroxycoumarins, secoiridoid glucosides, phenylethanoids and flavonoids. Biological studies reveal significant antimicrobial, antioxidative, photodynamic damage prevention, wound healing, anti-inflammatory, immunomodulatory and antiviral activities, and support the use of the bark in the folk medicine. PMID- 11429239 TI - Cytoprotective role of Solanum nigrum against gentamicin-induced kidney cell (Vero cells) damage in vitro. AB - The 50% ethanol extract of the whole plant of Solanum nigrum was tested in vitro for its cytoprotection against gentamicin-induced toxicity on Vero cells. Cytotoxicity was significantly inhibited as assessed by the Trypan blue exclusion assay and mitochondrial dehydrogenase activity (MTT) assay. The test extract also exhibited significant hydroxyl radical scavenging potential, thus suggesting its probable mechanism of cytoprotection. PMID- 11429240 TI - Anti-inflammatory and anti-ulcer activity of Calligonum comosum in rats. AB - The 10% ethanol extract of the aerial parts of Calligonum comosum (50--400 mg/kg, i.p.) significantly reduced the increase in hind paw oedema induced by carrageenan in rats. Following sub-acute oral administration of the extract (50 and 100 mg/kg, daily for 14 days), a significant anti-inflammatory activity in the cotton pellet model was observed. Pre-treatment with the extract (100, 200 and 400 mg/kg) produced a significant and dose-dependent inhibition to the acute gastric ulcers induced by phenylbutazone, indomethacin, 0.2 N NaOH and 80% ethanol. PMID- 11429241 TI - Studies on the anti-inflammatory properties of Entada abyssinica. AB - The defatted methanolic extract of Entada abyssinica was evaluated for anti inflammatory activity in acute and chronic models of inflammation. The extract (50--200 mg/kg, p.o.) exhibited dose-dependent and significant inhibition of both the carrageenan-induced rat paw oedema and granuloma tissue formation in rats. The extract (50--200 mg/kg, p.o.) was also found to inhibit the acetic acid induced vascular permeability in a dose-dependent fashion in mice. This study demonstrated the anti-inflammatory activity of Entada abyssinica. PMID- 11429242 TI - Evaluation of the anti-inflammatory and analgesic properties of Chasmanthera dependens leaf methanol extract. AB - A methanol extract of the dried leaves of Chasmanthera dependens was investigated for anti-inflammatory and analgesic activities. The extract (100--400 mg/kg, p.o.) produced dose-related inhibition of carrageenan-induced paw oedema and cotton pellet-induced granuloma in rats. Furthermore, an inhibition in the leakage of Evan's blue induced by acetic acid was observed in mice. At same doses, analgesic effect was also observed on writhing response induced by acetic acid as well as on the early and late phase of formalin-induced paw licking in mice. PMID- 11429243 TI - Wound healing activity of the aqueous extract of Thespesia populnea fruit. AB - Villagers have traditionally used the poultice prepared from the fruits of Thespesia populnea to treat a variety of skin ailments including wounds. In the present study, the aqueous extract of T. populnea fruit showed significant wound healing activity in the excision wound and incision wound models in rats following topical and oral administration, respectively. PMID- 11429244 TI - Cyclopeptide alkaloids from Zizyphus jujuba. AB - A new cyclopeptide alkaloid, jubanine-C (1), together with known alkaloids scutianine-C (4) and zizyphine-A (5), have been isolated from the stem bark of Zizyphus jujuba and identified by spectral analysis. PMID- 11429245 TI - Explaining photodermatosis: cyclopentenone vs. alpha-methylene-gamma-lactone natural products. AB - The possible role of cyclopentenone-containing sesquiterpene lactones in the cause of photochemical chronic actinic dermatitis (CAD) is examined in light of recent reports that the alpha-methylene-gamma-lactone group of these natural products forms 2+2 photoadducts with the DNA base thymine. Neither cyclopentenone nor tenulin (a cyclopentenone-containing sesquiterpene lactone) form such photoadducts with thymine either with sunlight or a UV lamp (300 nm). In contrast, alpha-methylenebutyrolactone readily forms the 2+2 photoadduct with thymine in sunlight. Thus, the photochemical role of the alpha-methylene-gamma lactone group (rather than cyclopentenone) is strongly implicated in the CAD disease. PMID- 11429246 TI - Hepatoprotective effect of lupeol and lupeol linoleate on tissue antioxidant defence system in cadmium-induced hepatotoxicity in rats. AB - Lupeol, a pentacyclic triterpene and its ester derivative, lupeol linoleate, were investigated for their possible hepatoprotective effect against cadmium-induced toxicity in rats. Cadmium intoxicated rats showed elevated levels of malondialdehyde (basal and induced), and decreased levels of antioxidants and antioxidising enzymes in the liver. The oral administration of triterpenes (150 mg/kg, once a day for 3 days before injection of cadmium chloride) changed the tissue redox system by scavenging the free radicals and by improving the antioxidant status of the liver. Lupeol linoleate had a better effect on the antioxidant status of the liver, when compared to lupeol. PMID- 11429247 TI - Prevention of 4-nitroquinoline 1-oxide-induced rat tongue carcinogenesis by garlic. AB - The present study was undertaken to examine the inhibitory effect of garlic (Allium sativum) on 4-nitroquinoline 1-oxide (4NQO)-induced tongue carcinogenesis in male rats, both in the initiation and post-initiation phases. Lipid peroxidation, reduced glutathione (GSH), glutathione peroxidase (GPx), and glutathione S-transferase (GST) were used to monitor the chemopreventive potential of garlic. Biochemical estimations were carried out on tumour and normal tongue tissues. Diminished lipid peroxidation in the tumour tissue was accompanied by a significant increase in the levels of GSH, GPx and GST. Administration of garlic (250 mg/kg, p.o., three times a week) effectively suppressed 4NQO-induced tongue carcinogenesis as revealed by the absence of carcinomas in the initiation phase and their reduced incidence in the post initiation phase. The results of the present study suggest that garlic may exert its chemopreventive effects by modulating lipid peroxidation and enhancing the levels of GSH, GPx and GST. PMID- 11429248 TI - Flavonol glycosides from Mentha lavandulacea. AB - In addition to kaempferol 3-O-rhamnoside-7-O-glucoside, the methanol extract of the aerial parts of Mentha lavandulacea yielded three new flavonoids identified as kaempferol 3-O-glucosyl-(1-->2)-rhamnoside-7-O-glucoside (1), kaempferol 3-O (6"'-p-coumaroylglucosyl)-(1-->2)-rhamnoside-7-O-glucoside (2) and kaempferol 3-O (4"'-p-coumaroylglucosyl)-(1-->2)-rhamnoside-7-O-glucoside (3). PMID- 11429249 TI - Antifeedant constituents from Fagara macrophylla. AB - Analysis of the polar fractions of an EtOH extract obtained from the bark of the African medicinal plant Fagara macrophylla led to the isolation and identification of the alkaloids oblongine (6), tembetarine (7) and magnoflorine (8) and the flavonoid hesperidin (9). These compounds, together with other metabolites (1--5) previously isolated from F. macrophylla, were tested for antifeedant activity in a binary-choice bioassay. The acridone alkaloid xanthoxoline (4) was found to have a potent antifeedant activity against larvae of both Spodoptera frugiperda and S. littoralis. 1-Hydroxy-3-methoxy-N-methyl acridone (2), arborinine (3), tembetarine (7) and magnoflorine (8) were antifeedant against S. frugiperda. PMID- 11429250 TI - Antibacterial activity of Jatropha multifida roots. AB - Hexane, ethyl acetate, chloroform and methanol successive extracts of Jatropha multifida yellow rootbark, red rootbark and rootwood effectively inhibited the growth of B. subtilis and S. aureus at concentration of 200 microg/disk. PMID- 11429251 TI - Antibacterial activity of Clausena heptaphylla. AB - The antibacterial activity of different crude extracts of Clausena heptaphylla leaves as well as three purified coumarins, obtained from the cold methanol extract, is reported. PMID- 11429252 TI - Antibacterial activity of Bergenia ciliata rhizome. AB - The methanol extract of Bergenia ciliata (tested at 200--1000 microg/disc) showed a wide spectrum of concentration-dependent antibacterial activity. PMID- 11429253 TI - A preliminary study on the analgesic activity of Grangea maderaspatana. AB - The methanol extract of the whole plant of Grangea maderaspatana showed a dose dependent analgesic activity. At doses of 1 and 3 g/kg, the extract significantly (P<0.001) inhibited acetic acid-induced writhing in mice by 50 and 80%, respectively. PMID- 11429254 TI - Constituents of Coriaria ruscifolia fruits. AB - Corianin (1) and ellagic acid 3,3'-dimethylether (2) were obtained from the methanol extract of powdered fruits of Coriaria ruscifolia. Biological screening of both compounds and of the methanol extract revealed slight antibacterial activity and cytotoxicity. PMID- 11429255 TI - Antibacterial spectrum of Hypericum hookerianum. AB - The antibacterial activity of the chloroform, acetone and methanol extracts of Hypericum hookerianum leaves and stems was evaluated. All extracts showed antibacterial activity against six different gram-positive and gram-negative bacteria, the methanol extracts exhibiting the maximum inhibitory activity at 400 microg/ml. PMID- 11429256 TI - Antimicrobial activity of Cassia alata. AB - The methanol extracts of leaves, flowers, stem and root barks of Cassia alata showed a broad spectrum of antibacterial activity. The activity was increased on fractionation (petrol, dichloromethane, ethyl acetate), the dichloromethane fraction of the flower extract being the most effective. No activity was shown against tested moulds. PMID- 11429257 TI - Antifungal constituent from Gordonia dassanayakei. AB - The hexane extract of the stem bark of Gordonia dassanayakei showed high antifungal activity against plant pathogenic fungi Curvularia sp., Colletotrichum gloeosporioides, Rhizoctonia solani, Corynespora cassiicola, and Fusarium sp. A compound responsible for the antifungal activity was isolated and identified as 3 formyl-2,4-dihydroxy-6-methylbenzoic acid 3-hydroxy-4-(methoxycarbonyl)-2,5 dimethylphenyl ester (1). PMID- 11429258 TI - Insect antifeedant and growth regulating activities of quassinoids from Samadera indica. AB - Four quassinoids, indaquassin C (1), samaderins C (2), B (3) and A (4), isolated from the seeds and bark of Samadera indica, were tested for insect antifeedant and growth regulatory activities against the tobacco cutworm, Spodoptera litura. Indaquassin C was the most effective antifeedant. Samaderin C increased pupal duration and induced pupal mortality. PMID- 11429259 TI - (+)-Totarol from Chamaecyparis nootkatensis and activity against Mycobacterium tuberculosis. AB - The isolation of (+)-totarol as active compound against Mycobacterium tuberculosis is reported from Chamaecyparis nootkatensis outerbark. PMID- 11429260 TI - Antimicrobial activity of Clematis papuasica and Nauclea obversifolia. AB - The methanol extracts of Clematis papuasica leaves and stem bark and of Nauclea obversifolia leaves, stem and root barks showed awide spectrum of antibacterial activity which was increased on fractionation (petrol, dichloromethane, ethyl acetate), the ethyl acetate fractions being in all cases the most effective. None of the extractives was active against tested moulds. PMID- 11429261 TI - Antimicrobial activity of Rhynchosia beddomei. AB - The petroleum ether and ethyl acetate extracts of Rhynchosia beddomei leaves showed inhibiting activity against some bacterial and fungal species at different concentrations. PMID- 11429262 TI - Constituents of the stem bark of Ormosia wayana. AB - The isolation of lupeol, beta-sitosterol acetate and three quinolizidinic alkaloids, (+)-lupanine, (-)-sparteine and (-)-3 beta-hydroxy-13 alpha-tigloyloxy lupanine (1), from the stem bark of Ormosia wayana is reported. PMID- 11429263 TI - Triterpenes from Licania licaniaeflora. AB - Nine triterpenoids with lupane, oleanane, and ursane skeleton have been isolated and characterized from the leaves of Licania licaniaeflora. The structural identification was based on (1)H and (13)C-NMR spectral data. PMID- 11429264 TI - 7-Epimer oxindole alkaloids of Cabucala cryptophlebia: their (13)C-NMR and CD data. AB - The isolation, (13)C-NMR and CD data of two 7-epimer oxindole alkaloids, 10,11 dimethoxyisomitraphylline (1) and 10,11-dimethoxymitraphylline (2) from Cabucala cryptophlebia are reported. PMID- 11429265 TI - Saponins from Strasburgeria robusta. AB - The isolation of three saponins, 24-hydroxytormentic acid ester glucoside (1), niga-ichigoside F1 (2) and niga-ichigoside F2 (3), from the stem bark of Strasburgeria robusta, an endemic plant from New Caledonia, is reported. PMID- 11429266 TI - 24-Methylenecycloartenone from Bhesa nitidissima. AB - Complete (1)H and (13)C-NMR assignments are reported for 24 methylenecycloartenone (1) isolated from the stem of Bhesa nitidissima. PMID- 11429267 TI - Phytochemical study of Mentha longifolia of Morocco. AB - A phytochemical study of Moroccan Mentha longifolia was carried out. The essential oil showed unusual relevant amounts of piperitenone oxide and piperitone oxide, which constitute a new chemotype. From the non-volatile extract, five flavonoids were isolated, among which 5,7,4'-trihydroxy-6,2',3' trimethoxyflavone (1) is reported for the first time. PMID- 11429268 TI - Ultrastructure of NADPH diaphorase-positive nerve fibers and their terminals in the rat cerebral arterial system. AB - To investigate how perivascular NO synthase (NOS)-containing nerves in the cerebral arterial system are involved in controlling the cerebral circulation, we observed the ultrastructure of NOS-containing nerve fibers and their terminals by means of nicotinamide adenine dinucleotide hydrogen phosphate-diaphorase (NADPH d) histochemistry. We also observed the correlation between NADPH-d stained perivascular nerves and the perivascular sympathetic nerves, by means of double staining with NADPH-d histochemistry and tyrosine hydroxylase (TH) immunohistochemistry at the light microscopic level. NADPH-d-positive nerve fibers showed dense distribution mainly in the rostral portion of the circle of Willis and proximal portions of its main branches, where some of the NADPH-d positive fibers coexisted with TH-positive fibers in a single nerve bundle. NADPH d-positive nerve fibers were unmyelinated and had close contact with NADPH-d negative myelinated and unmyelinated nerve fibers in a single nerve bundle, and NADPH-d-positive nerve terminals also existed closely with NADPH-d-negative nerve terminals. The number of NADPH-d-positive nerve terminals and their ratio to all other terminals were significantly higher in the rostral portion of the circle of Willis and the proximal portion of its branches, than the caudal portion of the circle of Willis and the distal portion of its branches. Nerve terminals were observed to locate within 250 nm from the basal lamina of arterial smooth muscle cells in the rostral portion of the circle of Willis and proximal portion of its branching arteries. The present observation confirmed that NOS-containing nerve fibers truly innervate the smooth muscle cells of the arterial wall in the circle of Willis and its main branches. Close contact between NADPH-d-positive and negative nerve fibers and terminals in these arterial portions may indicate that NOS-containing perivascular nerves may work to modulate the rest of the other perivascular nervous system, such as the sympathetic nerves, to regulate the homeostasis of the arterial tonus. PMID- 11429269 TI - Depletion of glial cell line-derived neurotrophic factor in substantia nigra neurons of Parkinson's disease brain. AB - The distribution of nerve growth factor (NGF), ciliary neurotrophic factor (CNTF), glial cell line-derived neurotrophic factor (GDNF), brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4 (NT-4) in substantia nigra pars compacta (SNc) of Parkinson's disease (PD) brains was investigated by immunofluorescence. Cases studied included four 69-77 year old neurologically normal male controls and four 72-79 year old male PD patients. Integrated optical densities (IODs) of immunofluorescence over individual neuromelanin-containing neurons and in areas of neuropil and the number of neurons on H & E stained adjacent sections were quantitated with the use of the BioQuant Image Analyzer. Data were statistically analyzed by ANOVA, including the unpaired two-tailed Student t-test and the Mann-Whitney test. The results showed 55.8% (P<0.0001) dropout of SNc neurons in PD brains compared to age-matched controls. Despite considerable neuronal dropout, immunofluorescent NTFs in the PD brains showed differential reductions that were consistent within the group as compared to age-matched controls: reductions were GDNF, 19.4%/neuron (P<0.0001), 20.2%/neuropil (P<0.0001); CNTF, 11.1%/neuron (P<0.0001), 9.4%/neuropil (P<0.0001); BDNF, 8.6%/neuron (P<0.0001), 2.5%/neuropil. NGF, NT-3 and NT-4 showed no significant differences within surviving neurons or neuropil. Since the depletion of GDNF both within surviving neurons and neuropil was twice as great as that of CNTF and BDNF and since the other NTFs showed no changes, GDNF, of the tested NTFs, is probably the most susceptible and the earliest to decrease in the surviving neurons of SNc. These observations suggest a role for decreased availability of GDNF in the process of SNc neurodegeneration in PD. PMID- 11429270 TI - Immunohistochemical localisation of neuronal nitric oxide synthase in the rainbow trout kidney. AB - The distribution of nitrergic nervous structures in the trout kidney was studied by peroxidase-linked ABC immunostaining procedures using a polyclonal antibody raised against the neuronal isoform of nitric oxide synthase. The nitrergic plexus reaches the kidney along the vasculature, mainly running with the postcardinal vein where nitrergic fibres, microganglia like cellular clusters and isolated neurones were detected. The atubular head-kidney only showed isolated nitrergic fibres close to the larger arteries. On the other hand, the collecting tubules, collecting ducts, large arteries and glomerular arterioles of the tubular middle and posterior trunks were innervated by nitrergic fibres even though immunoreactive neurones were also observed in close apposition to some tubular elements and large arteries. These results suggest that, according to morphofunctional differences between the fish and mammalian kidneys, nitrergic neural structures may be involved in the control of particular renal functions in the rainbow trout. PMID- 11429271 TI - The chemical architecture of the rat's periaqueductal gray based on acetylcholinesterase histochemistry: a quantitative and qualitative study. AB - The chemoarchitecture of the periaqueductal gray has been extensively studied, based on acetylcholinesterase reaction and comparing it to other chemical markers. We have divided the periaqueductal gray into four main longitudinal columns, namely dorsomedial, dorsolateral, lateral and ventrolateral. We also identified the dorsal midline column, the supraoculomotor cap and the juxta aqueductal ring. The acetylcholinesterase gave rise to a strong reaction in the outer half of the lateral column, the outer half of the dorsomedial column, the supraoculomotor cap and the ventral half of the juxta-aqueductal ring. This labeling was in part complementary to that of the NADPH diaphorase and allowed the lateral column to be differentiated from the ventrolateral column. However, the inner half of both lateral and ventrolateral columns displayed the same chemical properties including acetylcholinesterase, tyrosine hydroxilase and serotonin. Thus, from the chemical view, these inner halves should be considered as one different region. Finally, the juxta-aqueductal ring was composed of two clearly different halves, i.e. dorsal and ventral. The dorsal half did not show any clear differences from the above columns and was negative for acetylcholinesterase, NADPH diaphorase and tyrosine hydroxilase, while the ventral half was clearly different from the lateral and ventrolateral columns and displayed a positive reaction to all those chemical markers. From these results, we strongly suggest the use of acetylcholinesterase histochemistry as a tool for accurate parcellation of the periaqueductal gray. PMID- 11429272 TI - Origin of the noradrenergic innervation of the superior olivary complex in the rat. AB - In the rat, the superior olivary complex contains lateral and medial olivocochlear neurones, which respectively innervate two separate targets within the cochlea; the auditory afferents contacting the inner hair cells and the outer hair cells themselves. Previous double label immunohistochemical studies have shown that both lateral and medial olivocochlear neurones are contacted by noradrenergic nerve endings, and electrophysiological studies on in-vitro rat brain slices have demonstrated that noradrenaline exerts a direct, predominantly excitatory effect on medial olivocochlear neurones. In this paper, we have investigated the origin of the noradrenergic input to the superior olivary complex (SOC). A retrograde tracer, Fluorogold, was used to map the inputs to the SOC, and this was combined with immunofluorescent staining for dopamine-beta hydroxylase (DbetaH) to identify which of the afferent inputs was noradrenergic. These experiments showed small numbers of neurones double-stained for both Fluorogold and DbetaH in the A6 cell group (the locus coeruleus). In the A7 cell group, within and medial to the lateral lemniscus, numerous Fluorogold labelled and DbetaH positive neurones were found, but no neurones were seen that were double-labelled. In none of the other major noradrenergic cell groups were labelled Fluorogold neurones ever detected. To confirm the results obtained by retrograde tracer injections, anterograde tracer injections with biotinylated dextran amine were made in the locus coeruleus. This resulted in labelled fibres within all subdivisions of the superior olivary complex. These experiments indicate that the noradrenergic input to the olivocochlear neurones originates solely from the locus coeruleus. The small numbers of double-labelled neurones found in the locus coeruleus indicate a very divergent non-selective noradrenergic input to the SOC. PMID- 11429273 TI - Amyloid beta peptide levels and its effects on hippocampal acetylcholine release in aged, cognitively-impaired and -unimpaired rats. AB - Excessive extracellular deposition of amyloid beta (Abeta) peptide in neuritic plaques and degeneration of forebrain cholinergic neurones, which innervate the hippocampus and the neocortex, are the invariant characteristic features of Alzheimer's disease (AD). Studies of the pathological changes that characterize AD, together with several other lines of evidence, indicate that Abeta accumulation in vivo may initiate and/or contribute to the process of neurodegeneration observed in the AD brain. However, the underlying mechanisms by which Abeta peptide influences/causes degeneration of the basal forebrain cholinergic neurones in AD brains remain obscure. We reported earlier that nM concentrations of Abeta-related peptides, under acute conditions, can potently inhibit K+-evoked endogenous acetylcholine (ACh) release from the hippocampus and the cortex but not from striatum in young adult rats (J. Neurosci. 16 (1996) 1034). In the present study, to determine whether the effects of Abeta peptides alter with normal aging and/or cognitive state, we have measured Abeta1-40 levels and the effects of exogenous Abeta1-40 on hippocampal ACh release in young adult as well as aged cognitively-unimpaired (AU) and -impaired (AI) rats. Endogenous levels of Abeta(1-40) in the hippocampus are significantly increased in aged rats. Additionally, 10 nM Abeta1-40 potently inhibited endogenous ACh release from the hippocampus of the three groups of rats, but the time-course of the effects clearly indicate that the cholinergic neurones of AI rats are more sensitive to Abeta peptides than either AU or young adult rats. These results, together with earlier reports, suggest that the processing of the precursor protein of Abeta peptide alters with normal aging and the response of the cholinergic neurones to the peptide possibly varies with the cognitive status of the animals. PMID- 11429275 TI - Abstracts of the 22nd International Congress of Chemotherapy. Amsterdam, The Netherlands, June 30-July 3, 2001. PMID- 11429277 TI - Moving towards the next generation. AB - In most organisms, primordial germ cells are set aside from the cells of the body early in development. To form an embryonic gonad, germ cells often have to migrate along complex routes through and along diverse tissues until they reach the somatic part of the gonad. Recent advances have been made in the genetic analysis of these early stages of germ line development. Here we review findings from Drosophila, zebrafish, and mouse; each organism provides unique insight into the mechanisms that determine germ cell fate and the cues that may guide their migration. PMID- 11429278 TI - Command and control: regulatory pathways controlling invasive behavior of the border cells. AB - The invasiveness of cancer cells resembles the normal behavior of cells that migrate into surrounding tissues during development. For example, the border cells in the Drosophila ovary undergo a partial epithelial to mesenchymal transition and invade the neighboring cluster of germline cells, migrating to the oocyte border. Once there, they provide patterning information to the oocyte and produce an eggshell specialization known as the micropyle. Border cell migration has been subjected to extensive genetic analyses using a variety of screening approaches. Recent findings demonstrate that conversion of the border cells from a stationary group of epithelial cells to invasive cells requires integration of the activities of at least two transcriptional regulatory pathways. One such pathway requires the slbo gene, which encodes Drosophila C/EBP, a basic region/leucine zipper transcriptional activator that is required for elevated expression of a number of downstream targets, including DE-cadherin and focal adhesion kinase (FAK). An independent pathway requires the activity of the ecdysone receptor and a recently identified co-activator for the ecdysone receptor known as Taiman (abbreviated TAI, pronounced ti-maan', meaning too slow). Ecdysone is produced in the Drosophila ovary in response to adequate nutrition and is required for progression of oogenesis through stage 9, when border cell migration occurs. Border cells mutant for tai accumulate abnormally high levels of adhesion complexes at their surfaces, which may account for their inability to migrate. Thus border cell migration requires a differentiation program mediated by the C/EBP pathway, which is required for elevated expression of a number of proteins required for motility. In addition, migration requires a hormonal signal that relays information regarding nutritional status and appears to be required for regulation of the proper localization of some of the C/EBP targets. These findings suggest that steroid hormones can regulate cell motility relatively directly, independent of the effects on proliferation. This may contribute to the metastatic effects of steroid hormones on certain cancers and the inhibition of metastasis by steroid hormone antagonists such as tamoxifen. PMID- 11429279 TI - The early steps of neural crest development. AB - The neural crest is an intriguing cell population that gives rise to many derivatives which are all generated far from their final destinations. From its induction to the delamination of the cells, multiple signalling pathways converge to regulate the expression of effector genes, the products of which endow the cells with invasive and migratory properties reminiscent of those displayed by malignant cells in tumours. As such, the neural crest constitutes an excellent model to study cell migration. PMID- 11429280 TI - Segmental organization of neural crest migration. AB - Avian neural crest cells migrate on precise pathways to their target areas where they form a wide variety of cellular derivatives, including neurons, glia, pigment cells and skeletal components. In one portion of their pathway, trunk neural crest cells navigate in the somitic mesoderm in a segmental fashion, invading the rostral, while avoiding the caudal, half-sclerotome. This pattern of cell migration, imposed by the somitic mesoderm, contributes to the metameric organization of the peripheral nervous system, including the sensory and sympathetic ganglia. At hindbrain levels, neural crest cells also travel from the neural tube in a segmental manner via three migratory streams of cells that lie adjacent to even-numbered rhombomeres. In this case, the adjacent mesoderm does not possess an obvious segmental organization, compared to the somitic mesoderm at trunk levels. Thus, the mechanisms by which the embryo controls segmentally organized cell migrations have been a fascinating topic over the past several years. Here, I discuss findings from classical and recent studies that have delineated several of the tissue, cellular and molecular elements that contribute to the segmental organization of neural crest migration, primarily in the avian embryo. One common theme is that neural crest cells are prohibited from entering particular territories in the embryo due to the expression of inhibitory factors. However, permissive, migration-promoting factors may also play a key role in coordinating neural crest migration. PMID- 11429281 TI - Neuronal migration. AB - Like other motile cells, neurons migrate in three schematic steps, namely leading edge extension, nuclear translocation or nucleokinesis, and retraction of the trailing process. In addition, neurons are ordered into architectonic patterns at the end of migration. Leading edge extension can proceed at the extremity of the axon, by growth cone formation, or from the dendrites, by formation of dendritic tips. Among both categories of leading edges, variation seems to be related to the rate of extension of the leading process. Leading edge extension is directed by microfilament polymerization following integration of extracellular cues and is regulated by Rho-type small GTPases. In humans, mutations of filamin, an actin associated protein, result in heterotopic neurons, probably due to defective leading edge extension. The second event in neuron migration is nucleokinesis, a process which is critically dependent on the microtubule network, as shown in many cell types, from slime molds to vertebrates. In humans, mutations in the PAFAH1B1 gene (more commonly called LIS1) or in the doublecortin (DCX) gene result in type 1 lissencephalies that are most probably due to defective nucleokinesis. Both the Lis1 and doublecortin proteins interact with microtubules, and two Lis1-interacting proteins, Nudel and mammalian NudE, are components of the dynein motor complex and of microtubule organizing centers. In mice, mutations of Cdk5 or of its activators p35 and p39 result in a migration phenotype compatible with defective nucleokinesis, although an effect on leading edge formation is also likely. The formation of architectonic patterns at the end of migration requires the integrity of the Reelin signalling pathway. Other known components of the pathway include members of the lipoprotein receptor family, the intracellular adaptor Dab1, and possibly integrin alpha 3 beta 1. Defective Reelin leads to poor lamination and, in humans, to a lissencephaly phenotype different from type 1 lissencephaly. Although the action of Reelin is unknown, it may trigger some recognition-adhesion among target neurons. Finally, pattern formation requires the integrity of the external limiting membrane, defects of which lead to overmigration of neurons in meninges and to human type 2 lissencephaly. PMID- 11429282 TI - The selector gene cut represses a neural cell fate that is specified independently of the Achaete-Scute-Complex and atonal. AB - The peripheral nervous system (PNS) of Drosophila offers a powerful system to precisely identify individual cells and dissect their genetic pathways of development. The mode of specification of a subset of larval PNS cells, the multiple dendritic (md) neurons (or type II neurons), is complex and still poorly understood. Within the dorsal thoracic and abdominal segments, two md neurons, dbd and dda1, apparently require the proneural gene amos but not atonal (ato) or Achaete-Scute-Complex (ASC) genes. ASC normally acts via the neural selector gene cut to specify appropriate sensory organ identities. Here, we show that dbd- and dda1-type differentiation is suppressed by cut in dorsal ASC-dependent md neurons. Thus, cut is not only required to promote an ASC-dependent mode of differentiation, but also represses an ASC- and ato-independent fate that leads to dbd and dda1 differentiation. PMID- 11429283 TI - Pattern formation in the lateral line of zebrafish. AB - The lateral line of fish and amphibians is a sensory system that comprises a number of individual sense organs, the neuromasts, arranged in a defined pattern on the surface of the body. A conspicuous part of the system is a line of organs that extends along each flank (and which gave the system its name). At the end of zebrafish embryogenesis, this line comprises 7-8 neuromasts regularly spaced between the ear and the tip of the tail. The neuromasts are deposited by a migrating primordium that originates from the otic region. Here, we follow the development of this pattern and show that heterogeneities within the migrating primordium prefigure neuromast formation. PMID- 11429284 TI - Gene trap insertion reveals two open reading frames in the mouse SSeCKS gene: the form predominantly detected in the nervous system is suppressed by the insertion while the other, specific of the testis, remains expressed. AB - Scaffold proteins play an important role in regulating signal transduction by targeting kinases and phosphatases in close proximity to their relevant substrates. SSeCKS protein has been described as a protein kinase C and A (PKC/PKA) anchoring protein as well as a PKC substrate with a tumor suppressor activity. In this study, we report the generation, via gene trapping in embryonic stem cells of mice carrying an insertion in the mouse SSeCKS gene. Through the molecular analysis of the insertion site, we show that SSeCKS contains two alternative promoters directing the synthesis of mRNAs (P1- and P2-mRNA), encoding two different proteins, one of which would be a truncated form of the other. Interestingly, these RNAs are differentially expressed, P2 being found exclusively in the male germ line, while P1 exhibits a dynamic and wider pattern of expression during embryonic development and in the adult; its expression is predominant in the nervous system. Finally, we show that P1- but not P2-mRNA expression is abolished by the insertion and furthermore that mice homozygous for the mutation lack SSeCKS in all tissues except the male germ cells. Nevertheless and surprisingly, these mice do not exhibit any obvious phenotype. The functional implications of these observations are discussed. PMID- 11429285 TI - Differentiation of embryonic stem cell-derived dopaminergic neurons is enhanced by survival-promoting factors. AB - Here, we describe the generation of viable and dopamine-producing neurons derived from pluripotent mouse embryonic stem cells. Neurotrophic factors in combination with survival-promoting factors, such as interleukin-1beta, glial cell line derived neurotrophic factor, neurturin, transforming growth factor-beta(3) and dibutyryl-cyclic AMP, significantly enhanced Nurr1 and tyrosine hydroxylase (TH) mRNA levels, whereas En-1, mash-1 and dopamine-2-receptor mRNA levels were not upregulated. In parallel, mRNA levels of the anti-apoptotic gene bcl-2 were found to be upregulated at terminal stages. Double immunofluorescence analysis revealed increased numbers of TH- and dopamine transporter-, but not gamma-aminobutyric acid- and serotonin-positive neurons in relation to synaptophysin-labeled cells by survival-promoting factors. Moreover, high-performance liquid chromatography analysis showed detectable levels of intracellular dopamine. We conclude that survival-promoting factors enhance differentiation, survival and maintenance of dopaminergic neurons derived from embryonic stem cells. PMID- 11429286 TI - Branching and differentiation defects in pulmonary epithelium with elevated Gata6 expression. AB - The transcription factor GATA6 is expressed in the fetal pulmonary epithelium of the developing mouse lung and loss of function studies strongly suggested that it is required for proper branching morphogenesis and epithelial differentiation. We have further investigated the role of GATA6 in this process by utilizing a pulmonary epithelium specific promoter to maintain high levels of GATA6 protein during fetal lung development. Transgenic mice expressing Gata6 cDNA under the control of the human Surfactant Protein-C (SP-C) promoter were generated and their lungs were analyzed during fetal stages. Transgenic lungs exhibit branching defects as early as embryonic day (E) 14.5 and molecular analysis just before birth (E18.5) shows a lack of distal epithelium differentiation whereas proximal epithelium is unaffected. Electron microscopic analysis and glycogen staining confirm the lack of differentiation to mature Type II cells. Thus, elevated levels of GATA6 protein affect early lung development and in analogy to other GATA factors in other tissues, GATA6 also plays a crucial role in the terminal differentiation in this case of the distal pulmonary epithelium. PMID- 11429287 TI - Spatial regulation of DELTA expression mediates NOTCH signalling for segmentation of Drosophila legs. AB - The Notch (N) signalling pathway is recruited for segregation of cell fates in a number of Drosophila tissue types. We show here that N dependent segmentation of Drosophila legs is regulated by a dynamic pattern of expression of its ligand, DELTA (DL). During third larval instar and early stages of pupation, high levels of DL expression is seen in stripes of cells in the leg imaginal discs which later form the proximal borders of leg joints. These domains also displayed heightened Dl enhancer activity. During subsequent stages of pupation, following segmentation of the leg primordium, DL expression becomes uniform throughout these segments barring the joints. We further show that regulatory Dl mutations or mis-expression of DL abolish leg segmentation. Domains of N signalling for segmentation of legs of flies are thus set up by a stringent spatial regulation of expression of its ligand at the segment border. Further, a comparable role of DL in antennal development reveals a common paradigm of DL-N signalling for segmentation of appendages in flies. PMID- 11429288 TI - Signals from the ventral midline and isthmus regulate the development of Brn3.0 expressing neurons in the midbrain. AB - The vertebrate midbrain consists of dorsal and ventral domains, the tectum and tegmentum, which execute remarkably different developmental programs. Tectal development is characterized by radial migration of differentiating neurons to form a laminar structure, while the tegmentum generates functionally diverse nuclei at characteristic positions along the neural axis. Here we show that neurons appearing early in the development of the tectum are characterized either by the expression of the POU-domain transcription factor Brn3.0, or by members of the Pax and LIM families. Early neurons of the rostral tegmentum co-express Brn3.0 and Lim1/2, and caudal tegmental neurons express Islet1/2. Notochord tissue or Shh-transfected epithelial cells, transplanted to the developing tectum, suppress the development of tectal neurons, and induce the differentiation of multiple tegmental cell types. The distance from the midbrain hindbrain boundary (MHB) determines the specific markers expressed by the tegmental neurons induced in the tectum, and the transplantation of MHB tissue adjacent to the rostral tegmentum also induces caudal markers, demonstrating the role of MHB signals in determining the phenotype of these early midbrain neurons. Co-culture of isolated midbrain neuroepithelium with Shh-expressing cells demonstrates that Shh is sufficient to convert tectal neurons to a tegmental fate. In mice lacking Shh, Brn3.0- and Pax7-expressing neurons typical of the tectum develop throughout the ventral midbrain, and gene expression patterns characteristic of early tegmental development do not appear. PMID- 11429289 TI - Identification of a regulatory allele of teashirt (tsh) in Drosophila melanogaster that affects wing hinge development. An adult-specific tsh enhancer in Drosophila. AB - A cis-acting regulatory element defined herein is required to drive teashirt (tsh) expression in the regions of the developing adult that give rise to proximal wing and haltere tissues. Loss of this expression results in the fusion of the proximal structures of the wing and halteres to the thoracic cuticle. This represents the first description of a viable adult-specific regulatory allele of tsh with a visible phenotype, and it enlarges our understanding of the expression of tsh and its function during the development of the adult. PMID- 11429290 TI - Expression of the receptor tyrosine kinase genes, Ror1 and Ror2, during mouse development. AB - In mammals, the Ror-family receptor tyrosine kinases consist of two structurally related proteins, Ror1 and Ror2, characterized by the extracellular Frizzled-like cysteine-rich domain and membrane proximal kringle domains. As an attempt to gain insights into their roles in mouse development, expression patterns of Ror1 and Ror2 during early embryogenesis were examined and compared. Interestingly, at early stages, Ror1 and Ror2 exhibit similar expression patterns in the developing face, including the frontonasal process and pharyngeal arches, which are derived from cephalic neural crest cells. On the other hand, they exhibit different expression patterns in the developing limbs and brain, where the expression of Ror2 was detected broadly compared with that of Ror1. At a later stage, both genes are expressed in a similar fashion in the developing heart and lung, yet in a distinct manner in the brain and eye. PMID- 11429291 TI - Sequence analysis of zebrafish chondromodulin-1 and expression profile in the notochord and chondrogenic regions during cartilage morphogenesis. AB - Chondromodulin-I (ChM-I) is suggested in higher vertebrate systems to function as a key regulatory protein for cartilage development. To further understand the process of chondrogenesis and the function of ChM-I, we have cloned the zebrafish cDNA for chondromodulin-1 (chm1) and have mapped the chm1 gene locus. The expression profile of chm1 was determined during zebrafish embryonic development and compared to that of type II collagen (col2a1). Maternal chm1 transcripts were detected before midblastula transition and zygotic expression of chm1 was first observed in the notochord at the 10-somite stage. At later developmental stages, chm1 expression was detected in areas surrounding the otic vesicles, in the developing craniofacial cartilage elements, and in the chondrogenic region of the pectoral fins. PMID- 11429292 TI - Expression of Pcdh15 in the inner ear, nervous system and various epithelia of the developing embryo. AB - We previously determined that Protocadherin 15 (Pcdh15) is associated with the Ames waltzer mutation in the mouse. Here we describe where the Pcdh15 gene is expressed at specific times during mouse development using RNA in situ hybridization. The expression of Pcdh15 is found in the sensory epithelium in the developing inner ear, in Rathke's pouch, and broadly throughout the brain with the highest level of expression being detected at embryonic day 16 (E16). Pcdh15 transcripts are also found in the developing eye, dorsal root ganglion, and the dorsal aspect of the neural tube, floor plate and ependymal cells adjacent to the neural canal. Additionally, expression is also detected in the developing glomeruli of the kidney, surface of the tongue, vibrissae, bronchi of the lung, and in the epithelium of the olfactory apparatus, gut and lung. PMID- 11429293 TI - Expression analysis of the histamine H(3) receptor in developing rat tissues. AB - Endogenous histamine is involved in tissue growth and cell proliferation. In accordance with a putative function of the H(3) receptor in this mitogenic effect, we show that H(3)-receptor mRNAs are expressed together with those of the histamine-synthesizing enzyme in the embryonic liver and adipose tissue, and in various epithelia. Finally, we show that activation of recombinant H(3) receptors enhances MAP kinase activity. PMID- 11429294 TI - Zebrafish lunatic fringe demarcates segmental boundaries. AB - Cell interactions involving Notch signaling are required for the demarcation of tissue boundaries in both invertebrate and vertebrate development. Members of the Fringe gene family encode beta-1,3 N-acetyl-glucosaminyltransferases that function to refine the spatial localization of Notch-receptor signaling to tissue boundaries. In this paper we describe the isolation and characterization of the zebrafish (Danio rerio) homologue of the lunatic fringe gene (lfng). Zebrafish lfng is generally expressed in equivalent structures to those reported for the homologous chick and mouse genes. These sites include expression along the A-P axis of the neural tube, within the lateral plate mesoderm, in the presomitic mesoderm and the somites and in specific rhombomeres of the hindbrain; however, within these general expression domains species-specific differences in lfng expression exist. In mouse, Lfng is expressed in odd-numbered rhombomeres, whereas in zebrafish, expression occurs in even-numbered rhombomeres. In contrast to reports in both mouse and chicken embryos showing a kinematic cyclical expression of Lfng mRNA in the presomitic paraxial mesoderm, we find no evidence for a cyclic pattern of expression for the zebrafish lfng gene; instead, the zebrafish lfng is expressed in two static stripes within the presomitic mesoderm. Nevertheless, in zebrafish mutants affecting the correct formation of segment boundaries in the hindbrain and somites, lfng expression is aberrant or lost. PMID- 11429295 TI - Expression patterns of Wnt genes in mouse gut development. AB - Wnt signaling regulates cell fate decisions and cell proliferation during development and in adult tissues in both invertebrates and vertebrates. Here we describe the identification of Wnt genes, Wnt2a, 4, 5a, 5b, 6 and 11, expressed in mouse embryonic gut development. Each of these genes exhibits a characteristic and regional-specific expression pattern along the anterior-posterior axis of the digestive tube between embryonic day (E) 12.5 and 16.5 of embryonic development. The expression of Wnt5a is confined to the mesenchymal compartment, while expression of Wnt4 is found both in the intestinal epithelium and the mesenteric anlage. Wnt11 is expressed in the epithelium of esophagus and colon, but also in mesenchymal cells of the stomach. Wnt5b and Wnt6 exhibit restricted expression in the epithelium of the esophagus. A characteristic regionalized expression pattern is observed in the developing stomach. Wnt5a is expressed in the mesenchymal layer of the prospective gland region but becomes restricted to the tip of the gland region by E14.5. Wnt11 is highly expressed at the gastro-esophageal junctions, while Wnt4 is found in the epithelium lining the pyloric region of the stomach but not in the epithelium of the prospective gland region. PMID- 11429296 TI - Co-localization of KLF6 and KLF4 with pregnancy-specific glycoproteins during human placenta development. AB - Pregnancy-specific glycoproteins (PSGs) are major placental proteins essential for the maintenance of normal gestation. However, little is known about their gene expression regulation during placentation. It was previously demonstrated that the human core promoter binding protein recently renamed Kruppel-like factor (KLF) 6 binds to a highly conserved sequence within the PSG promoters and is mainly expressed in human term placenta. Here, we determined the expression pattern of the 13 other KLFs during human placental development. We demonstrate that eight KLFs exhibit specific expression patterns in human placental tissues and membranes, in favor of a functional cooperation of specific KLFs during placentation. In addition, we demonstrate that KLF6, KLF4 and PSG proteins are co expressed in same cell types of placental villi and membranes. This experimental evidence further strengthens the potential cross talk of both transcription factors for PSG gene regulation in vivo. PMID- 11429297 TI - Expression of two novel zebrafish iroquois homologues (ziro1 and ziro5) during early development of axial structures and central nervous system. AB - Previously, we reported a zebrafish iroquois gene, ziro3, and its expression during early embryogenesis (Mech. Dev. 87 (1999) 165). In the present study, we have isolated two novel zebrafish iroquois genes, ziro1 and ziro5, homologs of mouse Irx1 and mouse Irx5, respectively. The expression of both genes is initiated in dorsal neuroectoderm and mesoderm during gastrulation. Later, their expression appears in the central nervous system (CNS), excluding the telencephalon and most of the diencephalon. ziro1 expression is complementary to that of ziro3 in the notochord and later in the gut. In contrast, ziro5 expression mostly overlaps with that of ziro3. Interestingly, all three iroquois zebrafish genes are expressed in the notochord while only Irx3 is active in the mouse notochord. Their expression in later stages of embryogenesis was also compared. PMID- 11429300 TI - Postabortion psychological adjustment: are minors at increased risk? AB - PURPOSE: To assess whether younger adolescents experience greater adverse psychological outcomes after abortion than those aged 18-21 years, whether abortion places all adolescents at risk for negative sequelae, and what factors predict negative outcomes. METHODS: A total of 96 young women aged 14-21 years seeking counseling for unwanted pregnancies at four clinics completed questionnaires after counseling. These included the Beck Depression Inventory (BDI), an emotion scale, questions regarding sociodemographic and reproductive background, feelings about pregnancy, and decision-making. Sixty-three respondents were reinterviewed 4 weeks postabortion and completed the BDI, emotion scale, Spielberger State Anxiety Inventory, Rosenberg Self-esteem Scale, Impact of Events Scale, and Positive States of Mind Scale. Chi-squares and Student's t-tests were used to compare: (a) responses of adolescents under 18 years of age with those 18-21 years, (b) preabortion and postabortion responses, and (c) the current sample with other samples of adolescents. RESULTS: Adolescents under age 18 years were less comfortable with their decision, but showed no other differences compared with those aged 18-21 years. Both groups showed significant improvement in psychological responses postabortion. Postabortion scores did not differ significantly from those of other adolescent samples reported in the literature. Preabortion emotional state and perception of partner pressure predicted postabortion response. CONCLUSIONS: Despite its legal significance, age 18 years was not a meaningful cutoff point for psychological response to abortion in this sample. There was no evidence that abortion poses a threat to adolescents' psychological well-being. PMID- 11429301 TI - Do adolescents appreciate the risks of smoking? Evidence from a national survey. AB - PURPOSE: To evaluate whether adolescents understand the risks of smoking when they decide to start. Estimates of objective risks that can be compared with epidemiologic evidence suggest that adolescents overstate the risks. Ratings of personal risk suggest the opposite. METHODS: A nationally representative telephone survey of 300 14- to 22-year-old nonsmokers and 300 14- to 22-year-old smokers was conducted. Respondents estimated both objective and personal risks of smoking, and smokers reported their plans to quit. Objective estimates were compared with both epidemiologic evidence and personal ratings of risk. Regression procedures were used to assess relationships between different estimates of risk and between risk estimates and plans to quit. RESULTS: Two of the three objective estimates of risk revealed high proportions of misunderstanding. Over 40% of smokers and 25% of nonsmokers underestimated, or did not know, the likelihood of smoking-related death, and over 40% did not know, or underestimated, the number of years of life lost owing to smoking. Although young people overestimated lung cancer risk relative to objective data, these estimates are inflated by underestimation of the fatality of lung cancer and by overlap with other illnesses not included in objective risk measures. Young smokers exhibited optimism about personal risks of smoking regardless of their perceptions of objective risk. Both objective and personal measures of risk predicted plans to quit. CONCLUSIONS: Because perceptions of both personal and objective risks are related to plans to quit, antismoking messages should include evidence about risk, particularly to the individual smoker. PMID- 11429302 TI - Peers, schools, and adolescent cigarette smoking. AB - PURPOSE: To investigate the effects of popularity, best friend smoking, and cigarette smoking within the peer networks on current smoking of seventh- through 12th-grade students. These factors were examined for adolescents attending schools with varying rates of student cigarette smoking. METHODS: This study used data from the saturated school sample of National Longitudinal Study of Adolescent Health (Add Health), a nationally representative school-based sample. The sample for the present study was 2525 adolescents in Grades 7-12 who completed an in-school questionnaire and an in-home interview. Information from the in-school questionnaire was used to construct measures of school smoking prevalence and popularity. Using peer nominations from the in-home interview, best friend smoking, and peer network smoking exposure were constructed using the peers' own reports of their cigarette smoking. Multiple regression techniques were used to estimate the risk of current cigarette smoking as a function of popularity, best friend smoking, peer network smoking, and school smoking prevalence, and all first-order interactions between measures of peer and school smoking prevalence. RESULTS: Adjusting for age, gender, race/ethnicity, parent education, school, and availability of cigarettes in the home, the risk of current smoking was significantly associated with peer networks in which at least half of the members smoked [odds ratio (OR) = 1.91], one or two best friends smoked (OR = 2.00), and with increasing rates of school smoking prevalence (OR = 1.73). In addition, there was a significant interaction of popularity and school smoking prevalence such that risk of current smoking was somewhat greater among popular students in schools with high smoking prevalence than among popular students in schools with low smoking prevalence. CONCLUSION: Findings suggest that school environments are important contexts for understanding peer group influences on adolescent cigarette smoking. PMID- 11429303 TI - Relationship of sexual orientation to substance use, suicidal ideation, suicide attempts, and other factors in a population of homeless adolescents. AB - PURPOSE: To explore the relationship of sexual orientation and gender to four sets of factors: (a) family history, (b) incarceration, (c) substance use, and (d) depression and suicide, in a population of homeless adolescents. METHODS: A sample of homeless adolescents was recruited in Portland, Oregon and assessed using semi-structured interviews at baseline, three months and six months. A total of 532 youths (216 females and 316 males) provided data on sexual orientation and other variables. Heterosexual (n = 391) and non-heterosexual youths (n = 141) were compared on all sets of factors, primarily using logistic regressions. RESULTS: 44.9% of females identified as lesbian or bisexual, while only 13.9% of males identified as gay or bisexual. Gay, lesbian, bisexual, and "unsure" (GLBU) youths were less likely to have been in foster care or arrested, but were more likely to have spent time in a locked mental health treatment facility. More than one-third of all participants reported use of injection drugs. GLBU youths were more likely to have recently used amphetamines and to have injected drugs, however, gay-bisexual males were less likely to have recently used marijuana. GLBU status was associated with recent measures of depression and suicidal ideation, but not with lifetime measures. Associations of sexual orientation with several lifetime measures were different than with prospective measures, demonstrating the limitations of using lifetime measures rather than recent or prospective measures. CONCLUSIONS: This population of homeless adolescents appears to be higher in its high rate of injection drug use and the large proportion of females who identify as lesbian or bisexual than found in other studies. The high rates of depression and suicidal ideation, especially among GLBU youth, are of great concern. PMID- 11429304 TI - Adolescent pregnancy, infant mortality, and source of payment for birth: Alabama residential live births, 1991-1994. AB - PURPOSE: To assess potential impact of programs to prevent adolescent pregnancy on infant mortality. METHODS: Infants (n = 247,503) born alive to Alabama residents from 1991 to 1994 were identified from birth cohort files and linked to infant death records. Deliveries funded by Medicaid, private insurance, and self payers were studied separately. Relative risks (RR) and population attributable risks were calculated. The latter provided estimated percentages by which infant (<1 year) death risks would have decreased if adolescent (< or =19 years) pregnancies had been prevented. RESULTS: Infants of mothers <16 years of age were at higher risk of death than were infants of mothers aged 20 to 34 years (adults): the RRs were 4.1, 3.4, and 1.4 for self-payers, private insurance, and Medicaid groups, respectively. In the self-payer and private insurance groups, but not the Medicaid group, infants of mothers aged 17 to 19 years were more likely to die than were infants of adults. Infants from repeat adolescent pregnancies comprised nearly one-third of all infants born to adolescents, and in each payer-group were more likely to die than were infants of adults: The RRs were 4.9, 2.5, and 1.3 for self-payers, private-insurance, and Medicaid groups, respectively. Theoretically, preventing all adolescent pregnancies and preventing only repeat adolescent pregnancies would have reduced infant mortality in the total population by 8% and 4%, respectively. CONCLUSIONS: Programs to prevent first and subsequent adolescent pregnancies probably have little effect on infant mortality. Efforts to prevent adolescent pregnancies should not have a short-term goal of notably reducing infant mortality. PMID- 11429305 TI - Adolescent medicine faculty development needs. AB - OVERVIEW: Adolescent medicine faculty physicians were surveyed regarding training, career, and perceived needs. Most stated that training in teaching, research, and administration would be useful. Time spent in adolescent medicine activities was not associated with satisfaction, whereas having a mentor was. These issues should be considered when addressing faculty development needs. PMID- 11429306 TI - The AMA Residency Training in Adolescent Preventive Services Project: report of the working group. The American Medical Association. AB - PURPOSE: To improve resident education in provision of adolescent preventive health care. The American Medical Association (AMA) Residency Training in Adolescent Preventive Services Project Working Group convened to identify specific goals and objectives (G&Os) for pediatric and family medicine resident education in adolescent clinical preventive services and recommend strategies to achieve these G&Os. METHODS: Iterative review process involving members of the working group, nine experienced teaching faculty and 16 resident physicians from family medicine and pediatric training programs, and an advisory board. RESULTS: We achieved consensus on appropriate G&Os for pediatric and family medicine residency education in adolescent clinical preventive services. Faculty and residents expressed concerns about achieving G&Os because of challenges to implementing effective training and evaluation strategies. Suggestions for achieving G&Os included development of an adolescent clinical preventive services curriculum and evaluation program that could be adapted for use in a variety of training program structures. Faculty and residents anticipated the success of a training curriculum would be influenced by: (a) availability of adequate numbers of skilled teaching faculty; (b) availability of time and support for faculty development and teaching efforts; and (c) exposure of residents to adequate numbers of adolescent patients in settings where there are clear expectations for delivery of comprehensive preventive services. CONCLUSIONS: The AMA Residency Training in Adolescent Preventive Services Project Working Group presents G&Os for organizing training experiences in adolescent clinical preventive services in family medicine and pediatric residency training programs and recommends strategies to achieve these G&Os. PMID- 11429307 TI - Catalytically active polymers obtained by molecular imprinting and their application in chemical reaction engineering. AB - Molecular imprinting is a way of creating polymers bearing artificial receptors. It allows the fabrication of highly selective plastics by polymerizing monomers in the presence of a template. This technique primarily had been developed for the generation of biomimetic materials to be used in chromatographic separation, in extraction approaches and in sensors and assays. Beyond these applications, in the past few years molecular imprinting has become a tool for producing new kinds of catalysts. For catalytic applications, the template must be chosen, so that it is structurally comparable with the transition state (a transition state analogue, TSA) of a reaction, or with the product or substrate. The advantage of using these polymeric catalysts is obvious: the backbone withstands more aggressive conditions than a bio material could ever survive. Results are presented showing the applicability of a molecularly imprinted catalyst in different kinds of chemical reactors. It is demonstrated that the catalysts can be utilized not only in batch but also in continuously driven reactors and that their performance can be improved by means of chemical reaction engineering. PMID- 11429308 TI - Block-replacement mutagenesis for functional dissection of multiple transcription factor complexes. AB - We propose a simple method for investigation of roles of individual transcription factors in complexes of multiple factors bound to the same cis element. By block replacement mutagenesis, the whole cis element is replaced with a new one containing a binding site for a single factor. From the reporter activity of the mutant promoter construct, the importance of the individual factor in transcriptional activation is deduced. This approach allowed us to functionally identify SP1 as the most important PR1 binding transcription factor in the MN promoter. PMID- 11429309 TI - Controlling lipase enantioselectivity for organic synthesis. AB - Lipases are used frequently as chiral catalysts in the synthesis of various fine chemicals and intermediates. The increasing need of compounds with high stereochemical purity requires catalysts with an improved and controlled performance. This overview emphasizes some important aspects for the control of lipase enantioselectivity and some examples where the enantioselectivity has been altered or reversed are highlighted. However, in several of these cases the complete explanation for the altered or reversed enantioselectivity remains unclear and needs to be solved. Three different strategies (engineering of the reaction medium, the substrate molecule, and the enzyme) for exploring lipase enantioselectivity at a molecular level are discussed and summarized. These three different approaches represent powerful tools for understanding the molecular basis for lipase enantioselective catalysis and can guide the rational improvement and tailoring of catalyst performance. By combining approaches from chemistry and biology much is learnt about the most important parameters controlling lipase enantioselectivity for organic synthesis. PMID- 11429310 TI - Prediction of presence and absence of two- and three-amino-acid sequences of human monoamine oxidase from its amino acid composition according to the random mechanism. AB - Three probabilistic procedures are used to analyse the human monoamine oxidase B (MAO-B). (1) Each kind of two-, three- and multi-amino-acid sequences is counted along MAO-B from one terminal to the other and their frequencies and probabilities are calculated. (2) The amino-acid sequences in MAO-B are compared with the theoretical amino-acid sequences and the present or absent theoretical amino-acid sequences in MAO-B are determined. (3) The random principle is used to predict the frequencies of presence and absence of amino-acid sequences in MAO-B from its amino acid composition and the comparison is made between predicted and counted frequencies. The results show the random principle can predict the presence of 99 two-amino-acid sequences in MAO-B and the absence of 38 and 7506 kinds of theoretical two- and three-amino-acid sequences from MAO-B. PMID- 11429311 TI - Hemolytic uremic syndrome: how do factor H mutants mediate endothelial damage? AB - Hemolytic uremic syndrome is characterized by microangiopathic hemolytic anemia, thrombocytopenia and acute renal failure. Data from recent genetic analyses reveal a clear association between the complement regulator factor H and the atypical form of this fatal human disease. The clustering of the identified mutations in the C-terminus of factor H identifies a "hot spot" that is central to the pathogenesis of the disease. What are the possible biological and functional consequences of the identified mutations for the disease process and mechanisms of disease progression? PMID- 11429312 TI - A role for leptin in autoimmunity? PMID- 11429313 TI - Moving Ras in and out of lipid rafts. PMID- 11429314 TI - Motility is more important than [correction of that] stealth. PMID- 11429315 TI - Gob genes, mucus and asthma. PMID- 11429316 TI - IL-1 versus TNF in arthritis? PMID- 11429317 TI - A vaccine for colorectral cancer. PMID- 11429318 TI - Oligomeric antigen receptors: a new view on signaling for the selection of lymphocytes. AB - Antigen receptors on lymphocytes utilize different signaling modes to control the positive and negative selection of lymphocytes. In addition, these receptors have to detect the amount and affinity of the antigen and set delicate threshold values for the activation of lymphocytes. It is suggested that the antigen receptors on B and T-cells form oligomeric complexes, inside of which, signals can be processed and amplified in a manner that might explain the different signaling outputs of these receptors. PMID- 11429319 TI - Follicular dendritic cells: beyond the necessity of T-cell help. AB - Follicular dendritic cells (FDCs) are potent accessory cells for B cells, but the molecular basis of their activity is not understood. Several important molecules involved in FDC-B-cell interactions are indicated by blocking the ligands and receptors on FDCs and/or B cells. The engagement of CD21 in the B-cell coreceptor complex by complement-derived CD21 ligand on FDCs delivers a crucial signal that dramatically augments the stimulation delivered by the binding of antigen to the B-cell receptor (BCR). The engagement of Fc gamma receptor IIB (FcgammaRIIB) by the Ig crystallizable fragment (Fc) in antigen-antibody complexes held on FDCs decreases the activation of immunoreceptor tyrosine-based inhibition motifs (ITIMs), mediated by the crosslinking of BCR and FcgammaRIIB. Thus, FDCs minimize a negative B-cell signal. In short, these ligand-receptor interactions help to signal to B cells and meet a requirement for B-cell stimulation that goes beyond the necessity of T-cell help. PMID- 11429320 TI - Mac-1(+) myelopoiesis induced by CFA: a clue to the paradoxical effects of IFN gamma in autoimmune disease models. AB - The mechanisms accounting for the protective role of endogenous interferon gamma (IFN-gamma) in certain murine autoimmune disease models, versus a disease promoting role in others, have remained elusive. The protective effect of IFN gamma might be unique to models that rely on the use of complete Freund's adjuvant (CFA) and whose pathogenesis is predominantly driven by delayed-type hypersensitivity. In these models, IFN-gamma counteracts disease development by inhibiting CFA-induced proliferation of a pathogenically important Mac-1(+) cell population(s). This calls into question our usual conceptualization of the balance between innate and specific immunity in these models, as well as their clinical relevance, particularly when the role of IFN-gamma or related cytokines is considered. PMID- 11429321 TI - Th2 responses without atopy: immunoregulation in chronic helminth infections and reduced allergic disease. AB - The immune response to helminth infections has long been known to share key features with the allergic response. In particular, both are typified by enhanced T helper 2 (Th2) responses with high levels of interleukin-4 (IL-4), IL-5 and IL 13, accompanied by eosinophilia and abundant IgE production. Paradoxically, the geographical distribution of helminth parasitism and allergic disease is complementary rather than coincident. Thus, the question arises does the Th2 response to parasites protect or pre-empt the host from developing Th2-linked allergic manifestations? It is suggested that downregulatory immune mechanisms, which dampen the anti-parasite response, might benefit the host by blocking progression to atopic reactions. This is of relevance in explaining how the "hygiene hypothesis" might operate immunologically and in the design of therapeutics. PMID- 11429322 TI - MICA and MICB genes: can the enigma of their polymorphism be resolved? AB - The human MHC class I chain-related genes (MICA and MICB) are located within the HLA class I region of chromosome 6. Their organization, expression and products differ considerably from classical HLA class I genes. MIC proteins are considered to be markers of "stress" in the epithelia, and act as ligands for cells expressing a common activatory natural killer-cell receptor (NKG2D). Molecular models are now available for the MICA protein, both bound and complexed with NKG2D. MICA molecules appear to be highly flexible and polymorphic, although the functional relevance and implications of their polymorphism have yet to be fully discerned. PMID- 11429323 TI - Human-SCID mouse chimeric models for the evaluation of anti-cancer therapies. AB - The ability to engraft human tumors and human immunocompetent cells successfully in severe combined immunodeficient (SCID) mice has spawned the development and use of human-mouse chimeric models to evaluate anti-cancer therapies. The lack of standardization and many other potential pitfalls have contributed to the current controversy surrounding the reliability of these different models. Five frequently used SCID mouse models and their specific applications are summarized with the specific aim of providing an objective discussion of the strengths and limitations of each model, together with suggestions for overcoming some of the variabilities and for improving the design and use of future models. PMID- 11429324 TI - Dendritic cells as a tool to induce anergic and regulatory T cells. AB - The induction of antigen-specific T-cell tolerance in the thymus and its maintenance in the periphery is crucial for the prevention of autoimmunity. As well as their stimulatory functions, there is growing evidence that dendritic cells, acting as professional antigen-presenting cells, also maintain and regulate T-cell tolerance in the periphery. This control function is exerted by certain maturation stages and subsets of different ontogeny, and can be influenced by immunomodulatory agents. What is the current state of knowledge of the "immunoregulatory" properties of dendritic cells and how might tolerance inducing dendritic cells be relevant to therapeutic applications in humans? PMID- 11429325 TI - Ig heavy-chain gene revision: leaping towards autoimmunity. AB - B cells can revise their antigen receptors outside the confines of the bone marrow by secondary Ig gene rearrangements. Although the initial motivation to perform these revisions might be to silence a self-reactive specificity, those B cells that reinitiate the recombination process can perform a series of "leaping" rearrangements and inadvertently shift their receptor specificity towards autoimmunity. Heavy-chain receptor revision, coupled with other atypical rearrangements, might contribute to autoantibody production in systemic lupus erythematosus. PMID- 11429326 TI - Personal and outdoor nitrogen dioxide concentrations in relation to degree of urbanization and traffic density. AB - To assess differences in exposure to air pollution from traffic in relation to degree of urbanization and traffic density, we measured personal and home outdoor nitrogen dioxide (NO(2)) concentrations for 241 children from six different primary schools in the Netherlands. Three schools were situated in areas with varying degrees of urbanization (very urban, fairly urban, and nonurban) and three other schools were located near highways with varying traffic density (very busy, fairly busy, and not busy). Weekly averaged measurements were conducted during four different seasons. Simultaneously, indoor and outdoor measurements were conducted at the schools. Personal and outdoor NO(2) concentrations differed significantly among children attending schools in areas with different degrees of urbanization and among children attending schools in areas close to highways with different traffic densities. For the children living near highways, personal and outdoor NO(2) concentrations also significantly decreased with increasing distance of the home address to the highway. Differences in personal exposures between children from the different schools remained present and significant after adjusting for indoor sources of NO(2). This study has shown that personal and outdoor NO(2) concentrations are influenced significantly by the degree of urbanization of the city district and by the traffic density of and distance to a nearby highway. Because NO(2) can be considered a marker for air pollution from traffic, the more easily measured variables degree of urbanization, traffic density, and distance to a nearby highway can all be used to estimate exposure to traffic-related air pollution. PMID- 11429327 TI - Maternal mortality in the United States: where are we going and how will we get there? PMID- 11429328 TI - Mouth-to-mouth ventilation during cardiopulmonary resuscitation: word of mouth in the street versus science. PMID- 11429329 TI - The effects of vasopressin on systemic hemodynamics in catecholamine-resistant septic and postcardiotomy shock: a retrospective analysis. AB - We retrospectively investigated the effects of continuous arginine vasopressin (AVP) infusion on systemic hemodynamics, acid/base status, and laboratory variables in patients (mean age [mean +/- SD]= 66.3 +/- 10.1 yr) with catecholamine-resistant septic (n = 35) or postcardiotomy shock (n = 25). Hemodynamic and acid/base data were obtained before; 30 min after; and 1, 4, 12, 24, 48, and 72 h after the start of AVP infusion. Laboratory examinations were recorded before and 24, 48, and 72 h after the start of AVP infusion. For statistical analysis, a mixed-effects model was used. The overall intensive care unit mortality was 66.7%. AVP administration caused a significant increase in mean arterial pressure (+29%) and systemic vascular resistance (+56%), accompanied by a significant decrease in heart rate (-24%) and mean pulmonary arterial pressure (-11%) without any change in stroke volume index. Norepinephrine requirements could be reduced by 72% within 72 h. During AVP infusion, a significant increase in liver enzymes and total bilirubin concentration and a significant decrease in platelet count occurred. Arginine vasopressin was effective in reversing systemic hypotension. However, adverse effects on gastrointestinal perfusion and coagulation cannot be excluded. IMPLICATIONS: In this retrospective analysis, the influence of a continuous infusion of an endogenous hormone (arginine vasopressin) on systemic hemodynamics and laboratory variables was assessed in patients with vasodilatory shock unresponsive to conventional therapy. Arginine vasopressin was effective in reversing systemic hypotension. However, adverse effects on gastrointestinal perfusion and coagulation cannot be excluded. PMID- 11429330 TI - Atrial fibrillation after coronary artery bypass graft surgery is unrelated to cardiac abnormalities detected by transesophageal echocardiography. AB - Atrial fibrillation is a common complication of coronary artery bypass graft (CABG) surgery that is associated with adverse patient outcomes. We evaluated whether preexisting abnormalities of cardiac structure or function detected with transesophageal echocardiography (TEE) are prevalent in patients later developing atrial fibrillation after CABG surgery. TEE imaging was performed after induction of general anesthesia, but before primary CABG surgery, in 62 consecutive patients without cardiac valvular disease or preexisting atrial fibrillation. Measurements included left atrial diameter, left ventricular wall thickness, left ventricular end-systolic and end-diastolic dimensions and fractional area change. Pulsed-wave Doppler measurements of pulmonary venous and trans-mitral blood flow velocity were obtained. Continuous monitoring with telemetry electrocardiography for the development of atrial fibrillation was performed. Eighteen patients (29%) developed postoperative atrial fibrillation. There were no significant differences in left atrial or left ventricular TEE variables or pulsed-wave Doppler pulmonary venous flow measurements between patients with and without postoperative atrial fibrillation. After adjusting for age and duration of aortic cross-clamping, there were no differences in the transmitral Doppler diastolic filling variables between these same groups. These data suggest that atrial fibrillation commonly occurs after CABG surgery in the absence of atrial enlargement or Doppler-derived cardiac functional abnormalities. The data imply that the use of TEE immediately before surgery would be an insensitive means for routine identification of patients susceptible to this arrhythmia. IMPLICATIONS: Transesophageal echocardiography performed immediately before coronary artery bypass graft (CABG) surgery is not useful for prediction of susceptibility to develop atrial fibrillation postoperatively. Postoperative atrial fibrillation commonly occurs after CABG surgery in the absence of preoperative atrial enlargement or Doppler derived functional abnormalities. PMID- 11429331 TI - The effects of heparin, protamine, and heparin/protamine reversal on platelet function under conditions of arterial shear stress. AB - Platelet dysfunction contributes to blood loss after cardiopulmonary bypass. This study examined the antiplatelet effects of heparin, protamine, and varying heparin/protamine ratios in an in vitro physiologic model and further elucidated the mechanism of the antiplatelet and anticoagulant effects of protamine. We used the Clot Signature Analyzer (CSA(TM)), a system that analyzes coagulation in flowing whole blood, to test two aspects of platelet function, with different concentrations of heparin and protamine, under conditions simulating arterial flow: collagen-induced thrombus formation (CITF) under moderate shear and high shear platelet activation, platelet hemostasis time (PHT). In addition, platelet aggregometry, celite activated clotting time (Hepcon(TM) ACT), prothrombin time (PT), and partial thromboplastin time (PTT) were measured. Both PHT and the CITF were prolonged by heparin at 20 microg/mL, protamine at 20 and 40 microg/mL, and heparin/protamine ratios of 1:1 and 1:2, but not at 1:1.5. The Hepcon ACT was prolonged by heparin 20 microg/mL and protamine alone at 20 and 40 microg/mL, was normal at a ratio of 1:1, and was prolonged at 1:1.5 and 1:2. Protamine 80 microg/mL prolonged the PT and PTT. Dependency on thrombin, protein kinase C activation, and nonspecific charge effects were examined. The direct thrombin inhibitor D-phenylalanyl-L-prolyl-L-arginyl-chloromethyl ketone prolonged the PHT and ACT, but not the CITF, whereas the polycationic molecules polyarginine and polylysine prolonged the CITF, but not the PHT. The effect of protamine on the PTT, but not PT, could be shortened by the addition of excess phospholipid. Therefore, heparin inhibits both high shear collagen-independent and moderate shear collagen-dependent platelet activation; however, the latter is not mediated by its antithrombin activity. Protamine's antithrombin effect may explain its inhibition of platelet activation at high shear stress. Protamine's nonspecific charge effects are more important for inhibiting moderate shear collagen-induced platelet activation. IMPLICATIONS: This study suggests that protamine reversal of heparin's antiplatelet effect occurs within a narrow window because of the direct antiplatelet effects of protamine. Antithrombin effects may explain the inhibition of shear activation of platelets by both heparin and protamine. Nonspecific charge effects of protamine may explain the inhibition of collagen platelet activation in the presence of medium shear. PMID- 11429333 TI - The relationship between hirudin and activated clotting time: implications for patients with heparin-induced thrombocytopenia undergoing cardiac surgery. AB - Anticoagulation with recombinant hirudin (r-hirudin) (Refludan) has been suggested as an alternative to heparin for patients with heparin-induced thrombocytopenia requiring cardiac surgery. We sought to develop a modified activated coagulation time (ACT) that would allow quantification of the levels of r-hirudin required during cardiopulmonary bypass (CPB). Twenty-one patients scheduled for elective cardiac surgical procedures requiring CPB were enrolled in this IRB-approved study. R-hirudin was added to blood specimens obtained before heparin administration (before CPB) and 30 min after heparin neutralization with protamine (after CPB) to result in concentrations of 0, 2, 4, 6, 7, or 8 microg/mL. Kaolin/ACT and complete blood count measurements were assayed in native specimens (first 10 patients, Phase I) or in specimens mixed with equal volumes of commercial normal plasma (second 11 patients, Phase II). In Phase I, good (r(2) = 0.83) linear relationships between ACT values and r-hirudin concentrations (< or =4 microg/mL) were observed in specimens obtained before CPB. However, ACT values were markedly prolonged (P < 0.0001) by r-hirudin in specimens obtained after CPB, with ACT values generally exceeding the ACT's detection limit (>999 s) at hirudin concentrations >2 microg/mL. In patient specimens mixed with normal plasma (Phase II), ACT/hirudin relationships (i.e., hirudin/ACT slope values obtained with hirudin concentration < or =4 microg/mL) in the post-CPB period (0.022 +/- 0.004 microg. mL(-1). s(-1)) were similar (P = 0.47) to those (0.019 +/- 0.004 microg. mL(-1). s(-1)) obtained in the pre-CPB period. Accordingly, a significant relationship between normal plasma supplemented ACT values and predilution hirudin concentration was obtained in the post-CPB (hirudin = 0.039ACT - 4.34, r(2) = 0.91) period. Although our data demonstrate that the ACT test cannot be used to monitor hirudin during CPB, the addition of 50% normal plasma to post-CPB hemodiluted blood specimens yields a consistent linear relationship between hirudin concentration and ACT values up to a predilution concentration of 8 microg/mL. Plasma-modified ACT may be useful in monitoring hirudin anticoagulation during CPB. IMPLICATIONS: A modified activated clotting time test system that may be helpful in monitoring hirudin anticoagulation in patients with heparin-induced thrombocytopenia during cardiac surgery with cardiopulmonary bypass is described. PMID- 11429335 TI - The accuracy of coagulation tests during spinal fusion and instrumentation. AB - Patients undergoing major spine surgery may acquire a perioperative coagulopathy from dilution of coagulation factors and/or platelets or fibrinolysis. The mechanisms of the coagulopathy and role of coagulation testing during these procedures are poorly defined. Theoretically, coagulation tests could be used perioperatively to determine which patients are at risk for significant bleeding and guide transfusion therapy. We retrospectively evaluated the sensitivity, specificity, and accuracy of coagulation tests in predicting excessive surgical bleeding in 244 consecutive patients undergoing thoracic, lumbar, or sacral spinal fusion with or without instrumentation. Excessive bleeding was reported by the surgeon in 39 of the patients and was defined as recurrent microvascular bleeding despite adequate use of electrocautery and suture or decreased clot formation of blood pooled within the surgical field. Patients with excessive clinical bleeding sustained larger estimated blood losses than those with normal hemostasis. The total number of allogeneic red blood cells, platelets, and fresh frozen plasma units were also larger in patients with excessive bleeding noted during surgery. The intraoperative coagulation tests with the most sensitivity and specificity were the international normalized ratio (INR), prothrombin time (PT), and activated partial thromboplastin time (aPTT). The INR had a sensitivity of 94%, a specificity of 88%, and an accuracy of 0.9 at a value of 1.4 (normal, 0.8-1.2). The PT had a sensitivity of 90%, a specificity of 64%, and an accuracy of 0.73 at a value of 13.5 s (normal, 8.4-12.0 s). The aPTT had a sensitivity of 85%, a specificity of 64%, and an accuracy of 0.71 at a value of 30.9 s (normal, 23-37 s). The thromboelastogram values were of marginal use. We conclude that the INR, PT, and aPTT may be helpful in guiding transfusion therapy in patients undergoing major spine surgery. IMPLICATIONS: Patients undergoing major surgery to the spine often acquire a perioperative coagulopathy. The prothrombin time and activated partial thromboplastin time had the greatest sensitivity and specificity for predicting bleeding in major surgery of the spine. The test values that differentiated normal from excessively bleeding patients could be used to guide transfusion therapy during surgery. PMID- 11429336 TI - Somatosensory evoked potential monitoring during carotid endarterectomy in patients with a stroke. AB - The aim of our study was to assess the characteristics and feasibility of somatosensory evoked potential (SSEP) monitoring in patients who have had a stroke undergoing carotid endarterectomy. We retrospectively reviewed the medical and SSEP records of 204 patients. The patients were divided into two groups: Stroke (n = 65) and No-Stroke (n = 139). The amplitude and latency of the N20-P25 cortical complex on the ipsilateral side (surgical) were compared with the contralateral side in each group and between groups. Stroke patients showed asymmetry of their cortical waveforms; the ipsilateral N20-P25 baseline amplitude was 1.5 +/- 1.0 microv versus 1.9 +/- 1.2 microv for the contralateral (P = 0.001), for No-Stroke patients 2.0 +/- 1.1 microv versus 2.1 +/- 1.1 microv (P = 0.2). Forty-eight percent of Stroke patients had a ratio (ipsilateral/contralateral amplitude) of <1.0 +/- 0.2 compared with 26% for No Stroke patients (P = 0.01). There were no differences in latency measurements, in the incidences of significant SSEP changes (four Stroke, six No-Stroke) and immediate postoperative neurological deficits (two Stroke, six No-Stroke) between the two groups. Nine patients (three Stroke, six No-Stroke) had a decrease in ipsilateral N20-P25 amplitude >50% after cross-clamping, and had a shunt inserted. In conclusion, patients with a history of a stroke before surgery had a decrease in the amplitude of the ipsilateral cortical peak. There were no differences in the incidences of SSEP changes or neurological deficits. IMPLICATIONS: Patients who have had a preoperative stroke may show asymmetry of their cortical baseline somatosensory evoked potential waveforms; however, this does not interfere with the ability to use somatosensory evoked potential as a monitor during surgery. PMID- 11429337 TI - Reduced regional and global cerebral blood flow during fenoldopam-induced hypotension in volunteers. AB - Dopamine has a wide spectrum of receptor and pharmacologic actions that may affect cerebral blood flow (CBF). A new, selective dopamine-1 agonist, fenoldopam, is a potent systemic vasodilator with moderate alpha(2)-receptor affinity. However, the effects of fenoldopam on the cerebral circulation are undefined. We therefore hypothesized that infusion of fenoldopam would decrease mean arterial blood pressure (MAP) and might concurrently decrease CBF via vascular alpha(2)-adrenoreceptor activation in awake volunteers. We studied nine healthy normotensive subjects, using positron emission tomography to measure CBF in multiple cortical and subcortical regions of interest. In addition, bioimpedance cardiac output and middle cerebral artery blood flow velocity were determined during fenoldopam-induced hypotension. Three men and four women, aged 25-43 yr, completed the study. Fenoldopam infused at 1.3 +/- 0.4 microg. kg(-1). min(-1) (mean +/- SD) reduced MAP 16% from baseline: from 94 (89-100) mm Hg (mean [95% confidence interval]) to 79 [74-85] mm Hg (P < 0.0001). During the fenoldopam infusion, both cardiac output (+39%), and heart rate (+45%) increased significantly, whereas global CBF decreased from baseline, 45.6 [35.6-58.5] mL. 100 g(-1). min(-1), to 37.7 [33.9-42.0] mL. 100 g(-1). min(-1) (P < 0.0001). Despite restoration of baseline MAP with a concurrent infusion of phenylephrine, global CBF remained decreased relative to baseline values at 37.9 [34.0-42.3] mL. 100 gm(-1). min(-1) (P < 0.0001). Changes in middle cerebral artery velocity did not correlate with positron emission tomography-measured changes of CBF induced by fenoldopam, with or without concurrent phenylephrine. IMPLICATIONS: In awake volunteers with (presumably) intact cerebral autoregulation,fenoldopam-induced hypotension significantly decreased global cerebral bloodflow (CBF). Clinicians should be aware of these pharmacodynamic effects when choosing a vasodilator to control blood pressure, especially in situations where control of CBF, cerebral blood volume, and intracranial pressure are therapeutic priorities. PMID- 11429338 TI - Terlipressin for treating intraoperative hypotension: can it unmask myocardial ischemia? AB - IMPLICATIONS: After administration of terlipressin to treat hypotension related to induction of general anesthesia, profound hypertension occurred in association with myocardial ischemia and occlusion of the left anterior descending coronary artery. The authors emphasize cautious use of this drug because of such adverse events. PMID- 11429339 TI - Intracoronary thrombolysis and intraaortic balloon counterpulsation for the emergency treatment of probable coronary embolism after repair of an acute ascending aortic dissection. AB - IMPLICATIONS: This report shows that if diffuse coronary thromboembolism is encountered during ascending aortic dissection-repair, the option of combining single-bolus, intracoronary thrombolysis with intraaortic balloon counterpulsation should be considered. PMID- 11429340 TI - Anesthetic management of patients with Takayasu's arteritis: a case series and review. AB - Takayasu's arteritis is a rare, chronic progressive panendarteritis involving the aorta and its main branches. Anesthesia for patients with Takayasu's arteritis is complicated by their severe uncontrolled hypertension, end-organ dysfunction resulting from hypertension, stenosis of major blood vessels affecting regional circulation, and difficulties encountered in monitoring arterial blood pressure. Takayasu's arteritis is an uncommon disease and previous descriptions of the anesthetic management of patients with this disease have been limited to isolated case reports in the anesthetic literature, mostly in women undergoing cesarean delivery. We present our experience in this series of eight patients for various emergency and elective surgical procedures and review their perioperative problems and management. IMPLICATIONS: This case series describes the anesthetic problems and management of patients with pulseless disease. PMID- 11429341 TI - Can pediatric anesthesiologists detect an occluded tracheal tube in neonates? AB - To determine whether pediatric anesthesiologists can reliably detect occluded tracheal tubes, 18 pediatric anesthesiologists who were blindfolded and fitted with earplugs manually ventilated the lungs of 16 neonates. Consent was obtained from the parents of the neonates. All auditory signals from the monitors were silenced. Six conditions were studied (for 3 min each) in random order: three models of Ayre's t-piece with the Jackson Rees modification and two fresh gas flows (FGF) (2 and 6 L/min). During each condition, the tracheal tube was clamped at five predetermined but randomized times. The volume/pressure relationships of the three t-piece models were determined. Tube occlusions were detected more frequently at a low FGF (82%) than at a high FGF (64%) (P < 0.001). Experienced anesthesiologists (>8 yr experience) detected occlusions (83%) more frequently than less experienced (<2 yr experience) anesthesiologists (63%) (P < 0.027). There was no interaction between FGF and experience. The type of circuit did not affect the detection rate. We conclude that during isolated hand ventilation with the t-piece, pediatric anesthesiologists can detect >80% of occluded tubes provided they use a low FGF or have >8 yr experience, but only 60% of occluded tubes at high FGF or if they have <2 yr experience. IMPLICATIONS: Hand ventilation of the lungs in neonates has been used to detect changes in respiratory compliance, but laboratory models have failed to demonstrate its usefulness. We determined that pediatric anesthesiologists could detect 83% of tracheal tube occlusions in neonates if either the fresh gas flow was 2 L/minor the pediatric anesthesiologist was experienced (> 8 yr). PMID- 11429342 TI - The dose-response relationship for clonidine added to a postoperative continuous epidural infusion of ropivacaine in children. AB - Epidurally administered clonidine enhances the quality and duration of postoperative analgesia when it is used as an adjunct to local anesthetics in children. We investigated the dose-response relationship for epidural clonidine when added to a continuous postoperative epidural infusion of ropivacaine. By use of an observer-blinded design, 55 pediatric patients (1-4 yr old) were randomly given a postoperative epidural infusion of plain ropivacaine 0.1% 0.2 mg. kg(-1). h(-1) (Group R), ropivacaine 0.08% 0.16 mg. kg(-1). h(-1) plus clonidine 0.04 microg. kg(-1). h(-1) (Group RC1), ropivacaine 0.08% 0.16 mg. kg(-1). h(-1) plus clonidine 0.08 microg. kg(-1). h(-1) (Group RC2), or ropivacaine 0.08% 0.16 mg. kg(-1). h(-1) plus clonidine 0.12 microg. kg(-1). h(-1) (Group RC3). A clear dose response relationship could be identified for a continuous infusion of epidural clonidine, with clonidine dosages in the 0.08-0.12 microg. kg(-1). h(-1) range providing improved postoperative analgesia (reduced Children's Hospital of Eastern Ontario pain score, increased time to first supplemental analgesic demand, and a reduced total number of doses of supplemental analgesics during the first 48 h after surgery). Analgesia was improved without any signs of increased sedation or other side effects. The adjunct use of epidural clonidine in the dosage range of 0.08-0.12 microg. kg(-1). h(-1) appears effective and safe for use in children. IMPLICATIONS: The addition of clonidine (0.08-0.12 microg.kg( 1).h(-1))to a continuous epidural infusion of ropivacaine was found to improve postoperative pain relief in children. No clinically significant signs of sedation or other side effects were observed. PMID- 11429343 TI - Spinal anesthesia with tetracaine in 7.5% or 0.75% glucose in adolescents and adults. AB - To examine whether adolescents and adults might develop different anesthetic distribution and hemodynamic consequences after spinal injection of 0.5% tetracaine in 7.5% or 0.75% glucose, we studied 100 ASA I or II patients who were scheduled for elective surgery to the lower limb and fulfilled the following criteria: age between 13 and 16 yr (Adolescent group, n = 40) or between 25 and 74 yr (Adult group, n = 60); height between 155 and 180 cm; and body mass index between 18 and 32 kg/m(2). Patients in each group were then randomly divided into two equal subgroups to receive spinal anesthesia with 0.5% tetracaine in either 7.5% or 0.75% glucose with 0.125% phenylephrine at the L3-4 interspace. With patients in the supine horizontal position, neural block was assessed by cold, pinprick, and touch sensation and a modified Bromage scale after the injection of the study drug. The 7.5% glucose solution produced a significantly higher and faster spread of blockade in adolescents than in adults. In contrast, there were no differences in the levels of three sensory modalities between the two age groups after the 0.75% glucose solution, which produced a lower spread of blockade than the 7.5% glucose solution in either age group. Adolescents given the 0.75% glucose solution developed a smaller maximum decrease in systolic pressure than those given the heavier solution. We conclude that adolescents may develop an extensive level of blockade more easily and quickly than adults after intrathecal hyperbaric tetracaine, but that the difference may be reduced by using a less heavy solution. IMPLICATIONS: The influence of age on the characteristics of spinal anesthesia is still controversial. Our results show that adolescents develop blockade more extensively and quickly than adults after spinal anesthesia with 0.5%tetracaine in 7.5% glucose but not after the 0.75% glucose solution. PMID- 11429344 TI - A randomized trial of tranexamic acid to reduce blood transfusion for scoliosis surgery. AB - Pediatric patients who undergo posterior spinal fusion surgery to correct scoliosis often require multiple blood transfusions. Tranexamic acid is a synthetic antifibrinolytic drug that reduces transfusion requirements in cardiac surgery and total knee arthroplasty. We evaluated the efficacy of prophylactic tranexamic acid to reduce perioperative blood transfusion requirements in a prospective, double-blinded, placebo control study. Forty patients, 9-18 yr of age, were randomized to either tranexamic acid (initial dose of 10 mg/kg and infusion of 1 mg. kg(-1). h(-1)) or placebo (isotonic saline). Perioperative management was standardized. A uniform transfusion threshold for noncell saved red blood cells was 7.0 g/dL. The total amount of blood transfused in the perioperative period was significantly reduced in the Tranexamic group (P = 0.045). No thrombotic complications were detected in either group. The administration of prophylactic tranexamic acid in patients with scoliosis undergoing posterior spinal fusion surgery has the potential to reduce perioperative blood transfusion requirements. IMPLICATIONS: The administration of prophylactic tranexamic acid in patients with scoliosis who are undergoing posterior spinal fusion surgery has the potential to reduce perioperative blood transfusion requirements. PMID- 11429345 TI - The incidence of emergence agitation associated with desflurane anesthesia in children is reduced by fentanyl. AB - The rapid emergence and recovery from general anesthesia provided by desflurane is associated with a frequent incidence of emergence agitation in children. We sought to determine the mean effective dose of fentanyl that would significantly reduce the incidence of emergence agitation while preserving rapid recovery. Thirty-two children undergoing adenoidectomy received general anesthesia with desflurane and a dose of fentanyl (1.25, 1.87, 2.8, and 4.2 microg/kg) determined by the classic up-down method. Recovery characteristics, including time to extubation, recovery, hospital discharge, agitation, pain, and vomiting, were recorded. Demographics and recovery features were assessed by analysis of variance and Kruskal-Wallis tests. The mean effective dose of fentanyl to reduce agitation was calculated with the Dixon-Massey method to be 2.5 +/- 6.2 microg. There were no significant differences when treatment groups were compared for recovery criteria. Postoperative emesis occurred in 75% of patients. The results of this study demonstrate that a dose of 2.5 microg/kg of fentanyl is sufficient to prevent emergence agitation while preserving the rapid recovery associated with desflurane anesthesia in children undergoing adenoidectomy. IMPLICATIONS: A dose of 2.5 microg/kg of fentanyl prevents emergence agitation associated with desflurane anesthesia in children undergoing adenoidectomy without delaying emergence. PMID- 11429346 TI - The effect on the hemodynamic stability of varying calcium chloride administration during protamine infusion in pediatric open-heart patients. AB - IMPLICATIONS: We conducted a randomized study in 147 pediatric patients undergoing cardiopulmonary bypass to determine when there are any differences in hemodynamic effects if CaCl(2) 20 mg/kg and protamine 5mg/kg are mixed together and infused over 10 min versus administering half of the calcium dose (10 mg/kg) as a bolus followed by a 10-min infusion of protamine 5 mg/kg and CaCl(2) 10mg/kg. PMID- 11429347 TI - The successful management of thoracoscopic thoracic duct ligation in a compromised infant with targeted lobar deflation. AB - IMPLICATIONS: We describe the successful management of thoracoscopic thoracic duct ligation in a 2.5-kg compromised infant by using selective lobar-bronchial blockade. The technique reduces the risk of intraprocedural physiologic impairment, allowing the benefits of otherwise minimally invasive thoracoscopic procedures, even when the conditions of children are severely compromised. PMID- 11429348 TI - The management of preoperative anxiety in children: an update. PMID- 11429349 TI - The clinical and biochemical effects of propofol infusion with and without EDTA for maintenance anesthesia in healthy children undergoing ambulatory surgery. AB - We conducted this randomized, double-blinded, comparative, parallel-group study to determine whether adding EDTA to propofol would affect the clinical profile, calcium and magnesium homeostasis, or renal function in healthy children. After the induction of anesthesia with halothane, 69 ambulatory surgical patients (1 mo to <17 yr old), received propofol without EDTA (n = 33) or propofol with EDTA (n = 36). Blood samples were obtained for the measurement of ionized calcium, ionized magnesium, and laboratory indicators of renal function. Hemodynamic measurements, recovery, and adverse events were recorded. Propofol with EDTA produced no significant effects on clinical efficacy or renal function. Propofol and propofol EDTA produced a statistically significant decrease from baseline in serum concentrations of ionized calcium and magnesium during infusion (P<0.05), but with no apparent clinical effect. Hemodynamic measurements generally remained stable and were similar for both groups. Statistically significant changes in systolic blood pressure, mean arterial pressure, and heart rate were not considered clinically significant. Adverse events were mild or moderate. The addition of EDTA does not alter the clinical profile of propofol in pediatric ambulatory surgical patients. With or without EDTA, propofol is associated with a decrease in ionized calcium with no apparent clinical effect. IMPLICATIONS: The addition of EDTA does not alter the clinical profile of propofol in pediatric ambulatory surgical patients. With or without EDTA, propofol is associated with a decrease in ionized calcium with no apparent clinical effect. PMID- 11429350 TI - Fast-tracking after outpatient laparoscopy: reasons for failure after propofol, sevoflurane, and desflurane anesthesia. AB - IMPLICATIONS: In this study, although 41%-94% of the patients were fast-track eligible after laparoscopic surgery, only 35%-53% of the patients actually bypassed the postanesthesia care unit (PACU) because of anesthetic-related factors and surgical complications. Residual sedation was the most common anesthetic-related cause of failure to bypass thePACU. PMID- 11429351 TI - Adding ketamine to morphine for patient-controlled analgesia after major abdominal surgery: a double-blinded, randomized controlled trial. AB - In this double-blinded, randomized controlled trial we tested if the addition of ketamine to morphine for patient-controlled analgesia (PCA) resulted in improved analgesic efficacy and lower pain scores compared with morphine PCA alone after major abdominal surgery. Seventy-one patients were randomly allocated to receive either morphine 1 mg/mL (Group M) or morphine 1 mg/mL plus ketamine 1 mg/mL (Group MK) delivered via PCA after surgery. No other analgesics or regional blocks were permitted during the 48-h study period. Postoperatively there were no differences between the groups for subjective assessment of analgesic efficacy, pain scores at rest, and on movement, opioid consumption, or adverse events. Group MK patients performed worse in cognitive testing (P = 0.037). There was an increased risk of vivid dreaming in patients who received ketamine (relative risk = 1.8, 95% confidence interval 0.78-4.3). We conclude that small-dose ketamine combined with PCA morphine provides no benefit to patients undergoing major abdominal surgery. IMPLICATIONS: We performed a randomized, controlled trial comparing the use of ketamine and morphine with morphine alone to relieve pain after major abdominal surgery.Ketamine did not improve pain relief and merely increased side effects. PMID- 11429352 TI - Understanding the mechanisms by which isoflurane modifies the hyperglycemic response to surgery. AB - We studied the effect of anesthesia on the kinetics of perioperative glucose metabolism by using stable isotope tracers. Twenty-three patients undergoing cystoprostatectomy were randomly assigned to receive epidural analgesia combined with general anesthesia (n = 8), fentanyl and midazolam anesthesia (n = 8), or inhaled anesthesia with isoflurane (n = 7). Whole-body glucose production and glucose clearance were measured before and during surgery. Glucose clearance significantly decreased during surgery independent of the type of anesthesia. Epidural analgesia caused a significant decrease in glucose production from 10.2 +/- 0.4 to 9.0 +/- 0.4 micromol. kg(-1). min(-1) (P < 0.05), whereas the plasma glucose concentration was not altered (before surgery, 5.0 +/- 0.2 mmol/L; during surgery, 5.2 +/- 0.1 mmol/L). Glucose production did not significantly change during fentanyl/midazolam anesthesia (before surgery, 10.5 +/- 0.5 micromol. kg( 1). min(-1); during surgery, 10.1 +/- 0.5 micromol. kg(-1). min(-1)), but plasma glucose concentration significantly increased from 4.8 +/- 0.1 mmol/L to 5.3 +/- 0.2 mmol/L during surgery (P < 0.05). Isoflurane anesthesia caused a significant increase in plasma glucose concentration (from 5.2 +/- 0.1 mmol/L to 7.2 +/- 0.5 mmol/L) and glucose production (from 10.8 +/- 0.5 micromol. kg(-1). min(-1) to 12.4 +/- 1.0 micromol. kg(-1). min(-1)) (P < 0.05). Epidural analgesia prevented the hyperglycemic response to surgery by a decrease in glucose production. The increased glucose plasma concentration during fentanyl/midazolam anesthesia was caused by a decrease in whole-body glucose clearance. The hyperglycemic response observed during isoflurane anesthesia was a consequence of both impaired glucose clearance and increased glucose production. IMPLICATIONS: Epidural analgesia combined with general anesthesia prevented the hyperglycemic response to surgery by decreasing endogenous glucose production. The increased glucose plasma concentration in patients receiving fentanyl/midazolam anesthesia was caused by a decrease in whole-body glucose clearance. The hyperglycemic response observed during inhaled anesthesia with isoflurane was a consequence of both impaired glucose clearance and increased glucose production. PMID- 11429353 TI - Cardiopulmonary resuscitation performed by bystanders does not increase adverse effects as assessed by chest radiography. AB - Important adverse effects of bystander cardiopulmonary resuscitation (CPR) are well known. We describe the number of nonmedical professional CPR-related complications in patients surviving cardiac arrest, as assessed by chest radiograph. Within 2 yr, all consecutive patients admitted to the department of emergency medicine at a university hospital who had a witnessed, nontraumatic, normothermic cardiac arrest were studied. Radiologically evaluated adverse effects were compared with Mann-Whitney U-tests between patients who received bystander basic life support (Bystander group) and patients who did not receive bystander basic life support before advanced life support was started (ALS group). For assessment of bystander CPR-associated complications, chest radiographs were used. Of 224 patients, 173 were eligible. The median age was 58 yr (interquartile range, 51-71 yr), and 126 patients (73%) were men. The incidence of adverse effects associated with assisted-ventilation maneuvers and external chest compressions did not differ significantly between groups (severe gastric insufflation, 17% vs 18% between the Bystander group [n = 59] and the ALS group [n = 96], respectively; suspicion of aspiration, 22% vs 17%, respectively; soft tissue emphysema, 2% vs 1%, respectively; and serial rib fractures, 8% vs 8%, respectively). CPR administered by nonmedical personnel did not increase the number of life support-related adverse effects in patients surviving cardiac arrest as assessed by means of chest radiograph on admission. IMPLICATIONS: Complications related to cardiopulmonary bypass (CPR) are not increased when CPR is administered by nonmedical personnel, as assessed by chest radiograph. These data may be valuable in motivating lay people to perform basic life support. PMID- 11429354 TI - Maternal mortality during hospital admission for delivery: a retrospective analysis using a state-maintained database. AB - This study reports the overall age- and race-specific delivery mortality ratios from January 1984 to December 1997 and medical and demographic risk factors associated with maternal death during hospital admission for delivery. We performed a retrospective case control study using patient records from a state maintained anonymous database of all nonfederal Maryland hospitals that performed deliveries from 1984 to 1997. Variables studied included patient demographics and International Classification of Diseases, 9th Revision, Clinical Modification, diagnosis, and procedure codes. Mortality was the outcome variable. Of the 822,591 hospital admissions for delivery during the 14-yr study period, there were 135 deaths. The overall delivery mortality ratio was 16.4. The most common diagnoses associated with mortality during hospital admission for delivery included preeclampsia/eclampsia (22.2%), postpartum hemorrhage/obstetric shock (22.2%), pulmonary complications (14%), blood clot and/or amniotic embolism (8.1%), and anesthesia-related complications (5.2%). The identification of medical and demographic risk factors may have significant implications creating initiatives aimed at decreasing the public health burden associated with maternal mortality. IMPLICATIONS: This study reports the medical and demographic risk factors associated with maternal death during hospital admission for delivery by using a state-maintained database. This information could prove useful in the creation of initiatives aimed at decreasing the public health burden associated with maternal mortality. PMID- 11429355 TI - Fetal oxygen content is restored after maternal hemorrhage and fluid replacement with polymerized bovine hemoglobin, but not with hetastarch, in pregnant sheep. AB - We investigated the ability of hemoglobin-based oxygen carrying solutions (HBOCs) to alleviate fetal hypoxemia from maternal hemorrhage. Fifteen pregnant ewes (132 day gestational age) were hemorrhaged 20 mL/kg over 1 h; they were randomized to receive 20 mL/kg IV of HBOC, hetastarch (HTS), or autologous blood (BLD) (n = 5 each) over 30 min and were monitored for 2 h. Hemorrhage significantly (P < or = 0.05) decreased maternal mean blood pressure (from 98 to 48 mm Hg, median), arterial oxygen content (from 12.2 to 11.1 mL/dL), and fetal arterial oxygen content (from 8.1 to 3.9 mL/dL). Fluid replacement restored maternal blood pressure in all groups, although maternal oxygen content immediately returned to baseline only after BLD or HBOC. Maternal oxygen saturation decreased after HBOC (from 98% to 88%). Fetal oxygen content rapidly returned to baseline with either BLD (7.1 mL/dL) or HBOC (8.0 mL/dL) but was never restored with HTS (4.7 mL/dL), and, 60 min after fluid replacement, it was higher with HBOC (8.3 mL/dL) than with HTS (4.7 mL/dL). Fetal plasma-free hemoglobin did not change after HBOC. In conclusion, maternal fluid replacement with HBOC or BLD effectively restored fetal oxygenation, primarily by restoring maternal oxygen content, whereas HTS did not. IMPLICATIONS: Hemoglobin solutions eliminate many limitations of blood transfusions. Our results show that fluid replacement with either blood or a hemoglobin solution, compared with hetastarch, restored fetal oxygenation in pregnant ewes after hemorrhage. If applicable to women, these results suggest a potential for the use of hemoglobin solutions in obstetrics. PMID- 11429356 TI - The effects of uterine and umbilical blood flows on the transfer of propofol across the human placenta during in vitro perfusion. AB - The safety of using propofol in parturients is controversial, and little information is available on the factors that influence the placental transfer of propofol. In this study, we investigated the effects of uterine and umbilical blood flows on the placental transfer of propofol by using the dually perfused human placental cotyledon. Placental transfer was evaluated on the basis of the placental clearances at various uterine and umbilical flow rates. The placental transfer of propofol was significantly facilitated by the increased uterine flow rates over the range from 7.5 to 25 mL/min. The placental clearances of propofol were also dependent on the umbilical flow rates over the range from 0.5 to 4.0 mL/min. In contrast, the placental transfer of antipyrine was flow dependent when the umbilical flow rate was <2.0 mL/min and became permeability limited when it was >2.0 mL/min. No differences in either maternal or fetal venous concentrations of propofol were observed as umbilical flow rates varied from 0.5 to 4.0 mL/min, suggesting that an equilibration across the placenta occurs at low flow rates. These results indicate that fetal uptake of propofol can be profoundly altered by the changes in both uterine and umbilical blood flows observed in various pathophysiologic conditions and that lipid solubility greatly influences placental transfer of drugs. IMPLICATIONS: Uterine and umbilical blood flows are determinant features in controlling the placental transfer of propofol, and, therefore, changes in these variables would significantly affect the extent of fetal exposure to propofol. PMID- 11429357 TI - Hyperbaric spinal ropivacaine for cesarean delivery: a comparison to hyperbaric bupivacaine. AB - We evaluated the clinical efficacy and safety of spinal anesthesia with 0.5% hyperbaric ropivacaine compared with 0.5% hyperbaric bupivacaine for elective cesarean delivery. Sixty healthy, full-term parturients were randomly assigned to receive either 12 mg of 0.5% hyperbaric bupivacaine or 18 mg of 0.5% hyperbaric ropivacaine intrathecally. There were no significant differences in demographic or surgical variables or neonatal outcomes between groups. Onset time of sensory block to T10 or to peak level was later in the Ropivacaine group (P < 0.05). The median (range) peak level of analgesia was T3 (T1-5) in the Bupivacaine group and T3 (T1-4) in the Ropivacaine group. Time for sensory block to recede to T10 did not differ between groups. Duration of sensory block was shorter in the Ropivacaine group (188.5 +/- 28.2 min vs 162.5 +/- 20.2 min; P < 0.05). Complete motor block of the lower extremities was obtained in all patients. Ropivacaine also produced a shorter duration of motor blockade than bupivacaine (113.7 +/- 18.6 min vs 158.7 +/- 31.2 min; P < 0.000). The intraoperative quality of anesthesia was excellent and similar in both groups. Side effects did not differ between groups. Eighteen milligrams of 0.5% hyperbaric ropivacaine provided effective spinal anesthesia with shorter duration of sensory and motor block, compared with 12 mg of 0.5% hyperbaric bupivacaine when administered for cesarean delivery IMPLICATIONS: Eighteen milligrams of 0.5% hyperbaric ropivacaine provided effective spinal anesthesia with shorter duration of sensory and motor block, compared with 12mg of 0.5% hyperbaric bupivacaine when administered for cesarean delivery. PMID- 11429358 TI - Nalbuphine versus propofol for treatment of intrathecal morphine-induced pruritus after cesarean delivery. AB - In this prospective, randomized, double-blinded study, we compared the efficacy of nalbuphine and propofol for treating intrathecal morphine-induced pruritus after cesarean delivery. One-hundred-eighty-one parturients who developed moderate to severe pruritus after the administration of intrathecal morphine were randomly allocated into two groups. One group received 3 mg IV nalbuphine (n = 91), and the other received 20 mg IV propofol (n = 90). The improvement of pruritus and other adverse effects was determined at 10 min after study drug administration. The treatment success rate was higher in the Nalbuphine group than in the Propofol group (83% vs 61%; P < 0.001). Among the successfully treated patients, recurrence rates of moderate to severe pruritus within 4 h were not significantly different (nalbuphine 9% versus propofol 7%; P = 0.76). Other side effects, such as decreased analgesia, increased nausea, vomiting, increased sedation, pain on injection, and dizziness, were not significantly different between groups. Sedation and pain on injection, which were the two most common side effects, were minor and clinically inconsequential. IMPLICATIONS: Nalbuphine was superior to propofol for the treatment of intrathecal morphine-induced pruritus after cesarean delivery. PMID- 11429359 TI - Sodium nitroprusside compared with isoflurane-induced hypotension: the effects on brain oxygenation and arteriovenous shunting. AB - We compared sodium nitroprusside (SNP)-induced hypotension with 3% isoflurane induced hypotension with regard to brain tissue oxygen pressure (PtO(2)), middle cerebral artery (MCA) blood flow, and cerebral arteriovenous shunting. Eight dogs were anesthetized with 1.5% isoflurane. After a craniotomy, a probe was inserted into the left frontoparietal brain cortex to mea-sure tissue gases and pH. Blood flow was measured in a secondary branch of the MCA by a flowprobe. Measurements were made during baseline 1.5% isoflurane, during 1.5% isoflurane and SNP-induced hypotension or 3% isoflurane-induced hypotension to a mean pressure of 60-65 mm Hg, and during continued treatment with SNP or 3% isoflurane with blood pressure support to baseline levels with phenylephrine. Shunting was calculated from arterial, sagittal sinus, and tissue (indicating capillary) oxygen content. During hypotension with SNP, PtO(2) decreased 50%, and shunting increased 50%. During hypotension with 3% isoflurane, PtO(2) and shunting did not change. Blood pressure support increased PtO(2) and MCA flow during both SNP and 3% isoflurane treatment. These results show that SNP is a cerebrovasodilator but that hypotension will decrease PtO(2), probably because of an increase in arteriovenous shunting and a decrease in capillary perfusion. IMPLICATIONS: We measured brain arteriovenous shunting and tissue oxygen pressure(PtO(2))during a 40% decrease in blood pressure induced by sodium nitroprusside (SNP)or 3% isoflurane. Large-dose isoflurane maintainedPtO(2) with no change in shunting. SNP infusion decreasedPtO(2) 50%and increased shunting 50%. This suggests that SNP-induced hypotension decreases PtO(2) because of a decrease in capillary perfusion. PMID- 11429360 TI - A comparison of the transient hyperemic response test and the static autoregulation test to assess graded impairment in cerebral autoregulation during propofol, desflurane, and nitrous oxide anesthesia. AB - The transient hyperemic response (THR) test has been used to assess cerebral autoregulation in anesthesia and intensive care. To date it has not been compared with the static autoregulation test for assessing graded changes in cerebral autoregulation. We compared the two tests during propofol, desflurane, and nitrous oxide anesthesia. Seven subjects were studied. For the THR test, changes in the middle artery blood flow velocity were assessed during and after a 10-s compression of the ipsilateral common carotid artery. Two indices of autoregulation--THR ratio (THRR) and strength of autoregulation (SA)--were calculated. For the test of static autoregulation, changes in the middle cerebral artery flow velocity after a phenylephrine-induces increase in mean arterial pressure were assessed, and the static rate of regulation (sROR) was calculated. The tests were performed before induction and after equilibrium at 0.5 minimum alveolar anesthetic concentration (MAC) and then at 1.5 MAC of desflurane. THRR, SA and sROR decreased significantly (P < 0.001) at 0.5 MAC and then at 1.5 MAC desflurane. CHanges in THRR and SA reflected the changes in sROR with a sensitivity of 100%. IMPLICATIONS: When compared with the established test of static autoregulation, the transient hyperemic response test provides a valid method for assessing graded impairment in cerebral autoregulation. PMID- 11429361 TI - The effects of alpha-human atrial natriuretic peptide and milrinone on pial vessels during blood-brain barrier disruption in rabbits. AB - The effects of alpha-human atrial natriuretic peptide (HANP) and milrinone on cerebral pial vessels, especially during blood-brain barrier (BBB) disruption, are not clear. We studied topical HANP (10(-14), 10(-12), and 10(-10) M) or milrinone (10(-7), 10(-5), and 10(-3) M), and IV HANP (0.1, 0.2, and 1.0 microg. kg(-1). min(-1)) or milrinone (0.5, 5.0, and 20.0 microg. kg(-1). min(-1)) with or without hyperosmolar BBB disruption, using a rabbit cranial window preparation. At 10(-12) and 10(-10) M topical HANP produced significant arteriolar (16%, 20%, respectively), but no venular dilation. Topical milrinone (10(-3) M) produced significant arteriolar and venular dilation (21%, 8%, respectively). IV HANP produced no arteriolar or venular changes at any dose except during BBB disruption, when it caused a significant arteriolar (16%, 16%, and 17%, respectively), but no venular dilation. In contrast, IV milrinone caused small but significant arteriolar and venular dilation without BBB disruption (arterioles, 6%, 7% and 8%, respectively; venules, 6% at 20.0 microg. kg(-1). min(-1)). During BBB disruption, these responses to milrinone were similar. Although HANP and milrinone each have a direct vasodilator effect on arterioles, their systemic administration at clinical doses could induce different effects. BBB disruptive conditions could increase the response of pial vessels to systemically administered HANP. IMPLICATIONS: Although alpha-human atrial natriuretic peptide (HANP) and milrinone each have a direct vasodilator effect on cerebral pial arterioles, their systemic administration at clinical doses could have different effects and blood-brain-barrier disruptive conditions could alter the response of pial vessels to HANP, but not to milrinone. PMID- 11429362 TI - The effects of single or multiple injections on the volume of 0.5% ropivacaine required for femoral nerve blockade. AB - We compared the effects of using a single- or multiple-injection technique on the volume of 0.5% ropivacaine required to block the femoral nerve, in a prospective, randomized, blinded fashion in which 50 premedicated patients received a femoral nerve block with 0.5% ropivacaine by use of a nerve stimulator and either a single- (n = 25) or multiple- (n = 25) injection technique. Muscular twitches were elicited at < or =0.5 mA before anesthetic injection. The designated volume of local anesthetic was equally divided among contraction of the vastus medialis, vastus intermedius, and vastus lateralis for the multiple injections, or it was injected at the contraction of the vastus intermedius with motion of the patella for the single injection. The local anesthetic volumes were varied for consecutive patients by using an up-and-down staircase method; a blinded observer determined the adequacy of nerve blockade (loss of pinprick sensation in the medial, patellar, and lateral portions of the knee, with concomitant block of the quadriceps muscle) 20 min after injection. The mean (95% confidence interval) volume required for blocking the femoral nerve with the multiple-injection technique (14 [12-16] mL) was significantly smaller than that observed with the single injection (23 [20-26] mL) (P = 0.001). According to logistic regression analyses, the 95% effective volumes of ropivacaine required to block the femoral nerve within 20 min after injection were 29 and 21 mL with a single or multiple injection, respectively. We conclude that searching for multiple muscular twitches reduces the volume of 0.5% ropivacaine required to produce blockade of the femoral nerve. IMPLICATIONS: We evaluated the effects of using a single- or multiple-injection technique on the volume of 0.5% ropivacaine required to block the femoral nerve. The 95%effective concentration values for producing the same degree of sensory and motor blockade of the femoral nerve within 20 min after injection were 29 mL after elicitation of a patella twitch and 21 mL when the three main branches of the femoral nerve were identified, potentially leading to an important benefit for patients receiving peripheral nerve blocks. PMID- 11429363 TI - A comparison of 1% prilocaine with 0.5% ropivacaine for outpatient-based surgery under axillary brachial plexus block. AB - We compared the use of 1% prilocaine with 0.5% ropivacaine for axillary brachial plexus anesthesia in a double-blinded manner in day-stay patients to determine the better of the two local anesthetics in terms of onset time and duration of motor block. Sixty patients scheduled for outpatient upper-limb surgery were allocated randomly to receive either prilocaine (28 patients) or ropivacaine (32 patients) at a volume of 0.7 mL/kg. The brachial plexus was located with a plexus needle and nerve stimulator. By 20 min after injection of prilocaine or ropivacaine, there was no difference in analgesic effect. By this time, it was apparent whether or not a block was going to be adequate for surgery. Pain returned after a mean of 278 min (SD 111 min; range, 160-630 min) with prilocaine as compared with 636 min (SD 284 min; range, 210-1440 min) with ropivacaine. Analgesia use was similar in both groups. Duration of motor block with prilocaine was a mean of 254 min (SD 62 min; range, 130-385 min), as compared with 642 min (SD 199 min; range, 350-1080 min) with ropivacaine. We conclude that there is no clinically important difference between 1% prilocaine and 0.5% ropivacaine in time to onset of axillary brachial plexus block when they are injected in equal volumes. There is a significantly longer duration of action with ropivacaine, which may make it less suitable for day-stay upper-limb surgery because of the handicap from reduced muscle power. IMPLICATIONS: This study compares two local anesthetics to determine which is most suitable for day-stay upper-limb surgery under axillary brachial plexus block. Prilocaine 1% is more suitable than ropivacaine 0.5% because of a more prolonged duration of action of ropivacaine, although this could be useful in other circumstances. PMID- 11429364 TI - Amantadine, a N-methyl-D-aspartate receptor antagonist, does not enhance postoperative analgesia in women undergoing abdominal hysterectomy. AB - N-methyl-D-aspartate (NMDA) antagonists administered before surgery will improve postoperative analgesia, presumably by inhibiting spinal sensitization processes. However, current clinical formulations of NMDA antagonists either enable only an oral application (i.e., dextromethorphan) or are associated with psychotropic side effects, as with the IV delivery of ketamine. Because of its noncompetitive NMDA receptor antagonist characteristics, amantadine may improve postoperative analgesia when administered before surgically induced trauma. In this prospective, randomized clinical study, we examined whether female patients undergoing elective abdominal hysterectomy experienced less postoperative pain when IV amantadine was applied in comparison with placebo before the start of surgery. Thirty patients were randomly assigned to receive 500 mL saline IV before the induction of standardized general anesthesia in Group 1 (Control group) or, in a double-blinded manner, 200 mg amantadine IV in 500 mL saline in Group 2 (Treatment group). Postoperative pain control was provided via IV patient controlled analgesia with piritramide. During the first 48 h after tracheal extubation, pain perception was assessed by visual analog scales, and all analgesic requirements were documented. There were no significant differences between the two groups with respect to pain scores, postoperative analgesic requirements, and the incidence of side effects. Because of no differences in postoperative pain or opioid consumption, we conclude that a preoperative dose of 200 mg amantadine IV fails to enhance postoperative analgesia in patients undergoing elective abdominal hysterectomy. IMPLICATIONS: Because of no differences in postoperative pain or opioid consumption, we conclude that a preoperative dose of 200 mg amantadine IV fails to enhance postoperative analgesia in patients undergoing elective abdominal hysterectomy. PMID- 11429365 TI - The antinociceptive effect of the combination of spinal morphine with systemic morphine or buprenorphine. AB - We sought to analyze the mode of interaction of spinal morphine with systemic morphine or buprenorphine, administered in a wide range of antinociceptive doses. The study was performed on Sprague-Dawley rats by using a plantar stimulation test and isobolographic and fractional analyses of drug interaction. The isobolographic and fractional analyses demonstrated that intrathecal morphine interacted with subcutaneous morphine in a synergistic manner while producing a 50% or 75% antinociceptive effect. The sum of D(75) fractions was more than that for 50% antinociception, suggesting a less dramatic interaction. The combination with a maximal relative dose of systemic morphine (0.66:1) showed a maximal degree of supraadditivity. The interaction between spinal morphine and systemic buprenorphine was similar to that of morphine/morphine, although the supra additivity was not as pronounced. For the doses that produced a 50% antinociceptive effect, a synergistic interaction was observed only for the combination with a morphine/buprenorphine ratio of 1.33:1. When the relative amount of intrathecal morphine was decreased or increased, the effect became additive. At the doses that produced 75% antinociception, both combinations of morphine and buprenorphine demonstrated supraadditive interaction. IMPLICATIONS: Spinal morphine interacts with systemic morphine or buprenorphine in asupraadditive manner. This mode of interaction most probably results from the simultaneous activation of spinal and supraspinal antinociceptive systems. Supraspinal structures played a more important role in the antinociceptive effect of experimental combinations than structures of the spinal cord. PMID- 11429366 TI - Opioid-induced hyperalgesia and incisional pain. AB - Opioids occupy a position of unsurpassed clinical utility in the treatment of pain of many etiologies. However, recent reports in laboratory animals and humans have documented the occurrence of hyperalgesia when the administration of opioids is abruptly tapered or discontinued, a condition known as opioid-induced hyperalgesia (OIH). In these studies we documented that rats administered morphine (40 mg. kg(-1). day(-1) for 6 days) via subcutaneous osmotic minipumps demonstrated thermal hyperalgesia and mechanical allodynia for several days after the cessation of morphine administration. Additional experiments using a rat model of incisional pain showed that that attributable to OIH were additive with the hyperalgesia and allodynia that resulted from incision. In our final experiments we observed that if naloxone is administered chronically before incision then discontinued (20 mg. kg(-1). day(-1) for 6 days), the hyperalgesia and allodynia that result from hind paw incision was markedly reduced. In contrast, naloxone 1 mg/kg administered acutely after hind paw incision increased hyperalgesia and allodynia. We conclude that the chronic administration of exogenous opioid receptor agonists and antagonists before incision can alter the hyperalgesia and allodynia observed in this pain model, perhaps by altering intrinsic opioidergic systems involved in setting thermal and mechanical nociceptive thresholds. IMPLICATIONS: The chronic administration of opioids followed by abrupt cessation can lead to a state of hyperalgesia. In these studies we demonstrate that the hyperalgesia from opioid cessation and from hind paw incision are additive in rats. We suggest that failure to take into consideration preoperative opioid use can lead to excessive postoperative pain. PMID- 11429367 TI - Decreased tensile strength of an epidural catheter during its removal by grasping with a hemostat. AB - IMPLICATIONS: Our case suggests that gripping an epidural catheter with a hemostat during the removal might result in accidental breakage of the catheter. To demonstrate the effect of the use of a hemostat, tensile strengths of catheters were measured while they were being held with either a stainless steel or rubber-sleeved hemostat. PMID- 11429368 TI - The reversal of fentanyl-induced tolerance by administration of "small-dose" ketamine. PMID- 11429369 TI - Pioneers in epidural needle design. AB - IMPLICATIONS: In this article we discuss the development of epidural needles and the historical factors leading to their invention. The most popular needles are described and their inventors acknowledged. PMID- 11429370 TI - The carbon dioxide absorption capacity of Amsorb is half that of soda lime. AB - A new CO(2) absorbent, Amsorb (A), which does not contain monovalent bases, is ideal because it does not degrade volatile anesthetics to either Compound A (from sevoflurane) or carbon monoxide (from desflurane, enflurane, or isoflurane). The CO(2) absorption capacity of A, however, has not been investigated under clinical conditions. In this study, we compared the longevity (time to exhaustion) and CO(2) absorption capacity (the volume of CO(2) absorbed before CO(2) rebreathing occurs) of A under low-flow anesthesia (1 L/min) with those of two soda lime absorbents-Medisorb (M) and Sodasorb (S)-by using a 750-mL ADU canister and a 1350-mL Aestiva 3000 canister. In the study with the ADU canister, the longevity of A was 213 +/- 71 min, significantly less than those of M (445 +/- 125; P < 0.01) and S (503 +/- 89; P < 0.001). The CO(2) absorption capacity (L/100 g absorbent) of A was 5.5 +/- 1.2, significantly less than those of M (10.7 +/- 1.7) and S (12.1 +/- 1.8; P < 0.001). In the study with the Aestiva 3000 canister, the longevity of A was 218 +/- 61 min, significantly less than those of M (538 +/- 136) and S (528 +/- 103; P < 0.001). The CO(2) absorption capacity (L/100 g absorbent) of A was 7.6 +/- 1.6, significantly less than those of M (14.4 +/- 1.8) and S (14.8 +/- 2.3; P < 0.001). These results indicate that the CO(2) absorption capacity of A is half that of M or S and that the difference in the CO(2) absorption capacity between A and M or S is almost constant, regardless of the canister design. IMPLICATIONS: The CO(2) absorption capacity of Amsorb is half that of Medisorb and Sodasorb under clinical low-flow (1 L/min) anesthesia with either a 750-mL Ohmeda ADU compact or a 1350-mL Ohmeda Aestiva 3000 canister. PMID- 11429371 TI - Resting esophageal sphincter pressures and deglutition frequency in awake subjects after oropharyngeal topical anesthesia and laryngeal mask device insertion. AB - We investigated the effects of oropharyngeal topical anesthesia and placement of the standard (LMA) and the ProSeal (PLMA) laryngeal mask airway on resting gastroesophageal barrier pressure (GEBP), upper esophageal sphincter pressure (UESP), and deglutition frequency in awake subjects. Each subject was studied on 2 consecutive days: 1 day with the LMA and the other with the PLMA, in random order. GEBP and UESP were measured between deglutitions by using a pull-through technique in five sequential conditions: 1) after acclimatization to the manometer, 2) after topical anesthesia, 3) after the LMA or PLMA was self inserted and the cuff inflated with either 10 or 30 mL of air in random order, 4) after the cuff volume was adjusted to the other randomized volume, and 5) after LMA or PLMA removal. Deglutition frequency was determined between pressure measurements by using a neck microphone. UESP was always larger than GEBP (P < 0.001 for all). Topical anesthesia had no influence on GEBP, UESP, or deglutition frequency. LMA and PLMA placement did not influence GEBP or UESP, but deglutition frequency was higher (P < 0.02 for all). GEBP and UESP did not vary between devices for any condition. Cuff volume did not influence GEBP or UESP. Deglutition frequency was more frequent for the LMA than the PLMA at a 30-mL cuff volume (P = 0.008). We conclude that resting GEBP and UESP are unaffected by oropharyngeal topical anesthesia and the LMA or PLMA in awake subjects, but that deglutition frequency is increased by the LMA or PLMA. This may have implications for the incidence of regurgitation in these situations. IMPLICATIONS: Resting gastroesophageal barrier pressure and upper esophageal sphincter pressure are unaffected by oropharyngeal topical anesthesia and laryngeal mask devices in awake subjects, but deglutition frequency is increased by laryngeal mask devices. This may have implications for the incidence of regurgitation in these situations. PMID- 11429372 TI - A comparison of the relaxant effects of olprinone and aminophylline on methacholine-induced bronchoconstriction in dogs. AB - IV aminophylline, a nonselective phosphodiesterase (PDE) inhibitor, is often used to treat an asthma attack during anesthesia. However, in some instances, aminophylline-resistant attacks are observed. Selective PDE3 inhibitors are now clinically available and have been reported to produce bronchodilation. Thus, we compared the relaxant effects of olprinone, a novel PDE3 inhibitor, and aminophylline on methacholine-induced bronchoconstriction. Dogs were anesthetized with pentobarbital. Bronchoconstriction was elicited with methacholine (0.5 microg/kg + 5.0 microg. kg(-1). min(-1)) and assessed as percentage of changes in the bronchial cross-sectional area (BCA; basal = 100%) monitored by bronchoscope. Initially, the relaxant effects of olprinone (n = 8; 0-1000 microg/kg) and aminophylline (n = 8; 0-50 mg/kg) were compared. The bronchial cross-sectional areas were assessed before and 30 min after methacholine infusion began and 5 min after each dose of olprinone or aminophylline. We then determined whether propranolol (0.4 mg/kg) reversed the relaxation induced by olprinone (1000 microg/kg) and aminophylline (50 mg/kg). Olprinone and aminophylline dose dependently antagonized bronchoconstriction by 56.2% +/- 21.3% (SD) and 68.0% +/- 30.3% with -log 50% effective dose (mean) of 4.80 +/- 0.38 (15.8) microg/kg and 1.96 +/- 0.42 (10.9) mg/kg, respectively. Aminophylline 50 mg/kg significantly increased plasma epinephrine, whereas olprinone did not. In addition, propranolol significantly reduced aminophylline-induced relaxation, but not olprinone-induced relaxation. Therefore, the relaxant effects of olprinone are independent of plasma epinephrine, whereas aminophylline effects may partially result from increased circulating concentrations of epinephrine. IMPLICATIONS: We compared the relaxant effects of olprinone and aminophylline on methacholine-induced bronchoconstriction in dogs. The relaxant effects of olprinone are independent of plasma epinephrine, whereas the aminophylline effects may be partly caused by an increase in plasma epinephrine. PMID- 11429373 TI - The effect of temperature on solubility of volatile anesthetics in human tissues. AB - Hypothermia often occurs during surgery, a factor influencing anesthetic pharmacokinetics through its influence on solubility. Information on the tissue solubility of volatile anesthetics under hypothermia is limited. The present study supplies this information for the solubility of volatile anesthetics in human tissues. Tissue specimens of brain, heart, liver, muscle, and fat were obtained from 10 postmortem males (27 +/- 8 yr). Tissue/gas partition coefficients of desflurane, sevoflurane, enflurane, isoflurane and halothane were measured at 37 degrees C, 33 degrees C, 29 degrees C, 25 degrees C, 21 degrees C, and 17 degrees C. For each given tissue, the order of tissue/gas partition coefficient was halothane >enflurane >isoflurane >sevoflurane >desflurane. Tissue/gas partition coefficients at 37 degrees C differed significantly (P < 0.05) across drugs, except that liver/gas partition coefficients for isoflurane and enflurane did not differ. The logarithm of all tissue/gas partition coefficients increased linearly with decreasing temperature (P < 0.05). In conclusion, hypothermia increases tissue/gas partition coefficients of volatile anesthetics. The increases are proportional to those for blood/gas partition coefficients, and therefore tissue/blood partition coefficients will not change during hypothermic conditions. IMPLICATIONS: Volatile anesthetics are often used during hypothermic conditions, and tissue solubility of volatile anesthetics is an important determinant for the wash-in and washout of the anesthetics in tissue. Tissue/gas partition coefficients during hypothermia have implications for understanding the pharmacokinetics of volatile anesthetics at hypothermic conditions. PMID- 11429374 TI - A successful monitoring for intraoperative calcium stimulation test in complete resection of pancreatic insulinoma. AB - IMPLICATIONS: An intraoperative arterial stimulation, venous sampling method was recently used for resection of insulinoma. In using this technique, some anesthetic concerns complicate the usual management of insulinoma. We suggest methods for solving these problems. PMID- 11429375 TI - The implications of different failed endotracheal intubation rates. PMID- 11429376 TI - Esmolol is not an alternative to remifentanil for fast-track outpatient gynecologic laparoscopic surgery. PMID- 11429378 TI - Restoration of circulation after cessation of positive pressure ventilation in a case of "Lazarus syndrome". PMID- 11429377 TI - This little piggy went to MRI: the tale of the toe test. PMID- 11429380 TI - Video imaging of the larynx needs careful evaluation. PMID- 11429379 TI - Facilitation of pulmonary artery catheter placement by transesophageal echocardiography after tricuspid valve surgery. PMID- 11429381 TI - A new grip to help during endotracheal intubation. PMID- 11429382 TI - Cerebral oximetry and hyperbilirubinemia. PMID- 11429383 TI - Cardiac arrest during spinal anesthesia. PMID- 11429384 TI - Nitric oxide- and nitric oxide donors-induced relaxation and its modulation by oxidative stress in piglet pulmonary arteries. AB - Inhaled nitric oxide (iNO) is widely used in the treatment of pulmonary hypertension while inhaled NO donors have been suggested as an alternative therapy. The differential susceptibility to inactivation by oxidative stress and oxyhaemoglobin of NO and two NO donors, sodium nitroprusside (SNP) and S-nitroso N-acetyl-penicillamine (SNAP) were analysed in isolated endothelium-denuded pulmonary arteries from 2-week-old piglets stimulated with U46619. NO, SNAP and SNP relaxed the arteries (pIC(30)=7.73+/-0.12, 7.26+/-0.17 and 6.43+/-0.13, respectively) but NO was not detected electrochemically in the bath after the addition of SNP and only at concentrations at which SNAP produced more than 50% relaxation. The sGC inhibitor ODQ (10(-6) M) or the sarcoplasmic Ca(2+)-ATPase thapsigargin (2x10(-6) M) markedly inhibited the relaxation induced by NO, SNAP and SNP. Addition of oxyhaemoglobin (3x10(-7) M) or diethyldithiocarbamate (1 mM) markedly inhibited NO- (pIC(30)=6.88+/-0.07 and 6.92+/-0.18, respectively), weakly inhibited SNAP- and had no effect on SNP-induced relaxation. Xanthine oxidase (5 mu ml(-1)) plus hypoxanthine (10(-4) M) markedly inhibited NO- (pIC(30)=6.96+/-0.12) but not SNAP- or SNP-induced relaxation. Superoxide dismutase (SOD), MnCl(2), diphenileneiodonium and exposing the luminal surface of the rings outwards (inversion) potentiated the relaxant responses of NO (pIC(30)=8.52+/-0.16, 8.23+/-0.11, 8.01+/-0.11 and 8.20+/-0.10, respectively). However, SOD did not modify the NO detected by the electrode and had no effect on SNAP- or SNP-induced relaxation. Therefore, the kinetics and local distribution of NO release of NO donors influence the susceptibility to the scavenging effects of oxyhaemoglobin and superoxide. PMID- 11429385 TI - Electrophysiological effects of dronedarone (SR 33589), a noniodinated amiodarone derivative in the canine heart: comparison with amiodarone. AB - The electrophysiological effects of dronedarone, a new nonionidated analogue of amiodarone were studied after chronic and acute administration in dog Purkinje fibres, papillary muscle and isolated ventricular myocytes, and compared with those of amiodarone by applying conventional microelectrode and patch-clamp techniques. Chronic treatment with dronedarone (2x25 mg(-1) kg(-1) day p.o. for 4 weeks), unlike chronic administration of amiodarone (50 mg(-1) kg(-1) day p.o. for 4 weeks), did not lengthen significantly the QTc interval of the electrocardiogram or the action potential duration (APD) in papillary muscle. After chronic oral treatment with dronedarone a small, but significant use dependent V(max) block was noticed, while after chronic amiodarone administration a strong use-dependent V(max) depression was observed. Acute superfusion of dronedarone (10 microM), similar to that of amiodarone (10 microM), moderately lengthened APD in papillary muscle (at 1 Hz from 239.6+/-5.3 to 248.6+/-5.3 ms, n=13, P<0.05), but shortened it in Purkinje fibres (at 1 Hz from 309.6+/-11.8 to 287.1+/-10.8 ms, n=7, P<0.05). Both dronedarone (10 microM) and amiodarone (10 microM) superfusion reduced the incidence of early and delayed afterdepolarizations evoked by 1 microM dofetilide and 0.2 microM strophantidine in Purkinje fibres. In patch-clamp experiments 10 microM dronedarone markedly reduced the L-type calcium current (76.5+/-0.7 %, n=6, P<0.05) and the rapid component of the delayed rectifier potassium current (97+/-1.2 %, n=5, P<0.05) in ventricular myocytes. It is concluded that after acute administration dronedarone exhibits effects on cardiac electrical activity similar to those of amiodarone, but it lacks the 'amiodarone like' chronic electrophysiological characteristics. PMID- 11429386 TI - Role of protein kinase C in the reduction of infarct size by N-methyl-1 deoxynojirimycin, an alpha-1,6-glucosidase inhibitor. AB - Preischaemic treatment with N-methyl-1-deoxynojirimycin (MOR-14), an alpha-1,6 glucosidase inhibitor, attenuates glycogenolysis and lactate accumulation during ischaemia and markedly reduces infarct size in rabbit hearts. In the present study, we have investigated whether protein kinase C (PKC), a principal mediator of ischaemic preconditioning, is also involved in the cardioprotective effect of MOR-14. To assess the effect of PKC inhibition on infarct size in MOR-14-treated hearts, 38 rabbits were subjected to 30 min of ischaemia followed by 48 h of reperfusion. Infarct size, as a per cent of area at risk, was significantly smaller in rabbits administered 100 mg kg(-1) of MOR-14 10 min before ischaemia (17+/-2%, n=10), than in a control group (46+/-5%, n=10). This beneficial effect of MOR-14 was abolished when 5 mg kg(-1) of chelerythrine, a PKC inhibitor, was given 10 min prior to MOR-14 injection (39+/-4%, n=10), although chelerythrine alone did not alter infarct size (43+/-4%, n=8). Further, chelerythrine had no effect on MOR-14-induced attenuation of glycogen breakdown and lactate accumulation in hearts excised at 30 min of ischaemia. Immunoblot analysis of PKC in homogenates of Langendorff-perfused rabbit hearts revealed that MOR-14 significantly increased levels of PKC-epsilon in the particulate fraction at 20 and 30 min of ischaemia and in the cytosolic fraction at 30 min of ischaemia. Taken as a whole, our data suggest that PKC acts downstream of the inhibition of glycogenolysis by MOR-14 to reduce infarct size. Thus, activation of PKC is a more direct mediator of the cardioprotection afforded by MOR-14 than is inhibition of glycogenolysis. PMID- 11429387 TI - Generation of 8-epi-prostaglandin F(2alpha) in isolated rat kidney glomeruli by a radical-independent mechanism. AB - Isoprostanes comprise a group of free radical-catalyzed products of arachidonic acid. However, there is recent evidence pointing towards an enzyme-dependent formation of isoprostanes. With the use of isolated rat glomeruli we addressed the mechanisms of isoprostane generation. Synthesis of prostanoids and isoprostanes, including 8-epi-PGF(2alpha), was studied under conditions favouring radical formation. Cultured glomeruli formed different prostanoids including 8 epi-PGF(2alpha). Upon LPS challenge cyclo-oxygenase (COX)-2 expression was enhanced, and this was paralleled by a 2 - 9-fold increase in prostanoid formation, including isoprostanes. Addition of COX-isoform unselective inhibitors (diclofenac, indomethacin) or a selective inhibitor (NS-398) suppressed the synthesis of prostanoids, 8-epi-PGF(2alpha) and total isoprostane fraction; however, inhibition of the latter was less pronounced. Antioxidants such as butylated hydroxytoluene (BHT), nordihydroguaiaretic acid (NDGA), or dimethylurea exhibited an only minimal inhibitory effect on 8-epi-PGF(2alpha) synthesis. Moreover, ROS-generating drugs (menadione, methylviologen) or NADPH-driven radical formation were unable to cause the generation of significant amounts of 8 epi-PGF(2alpha) by rat glomeruli. In contrast, the total isoprostane fraction could be increased by menadione addition. These data provide further evidence for a radical-independent, but COX-dependent formation of 8-epi-PGF(2alpha) in renal tissue. Regarding the other isoprostanes, both radicals and COX enzymes contribute to their formation. Based on our data we assume that elevated release of vasoactive 8-epi-PGF(2alpha) has to be expected under conditions when the prostanoid system in the kidney is stimulated, e.g. under inflammatory conditions. Regarding renal oxidative injuries, the usefulness of 8-epi PGF(2alpha) as a representative marker molecule of oxidative stress has to be questioned. PMID- 11429388 TI - Straight-chain alcohols exhibit a cutoff in potency for the inhibition of recombinant glutamate receptor subunits. AB - The effects of n-alcohols (methanol to 1-decanol) on kainate-activated AMPA receptor subunit GluR1 and GluR3 ion currents were studied in Xenopus oocytes using the two-electrode voltage-clamp recording technique. For short-chain alcohols from methanol to 1-hexanol, potency for inhibition of GluR1 and GluR3 receptor-mediated current increased in proportion to the chain length or hydrophobicity of the alcohol. The IC(50) values of these alcohols for GluR1 were: methanol, 702 mM; ethanol, 170 mM; 1-propanol, 69 mM; 1-butanol, 20 mM; 1 pentanol, 17 mM; and 1-hexanol, 10 mM. For GluR3, IC(50) values were: methanol, 712 mM; ethanol, 238 mM; 1-propanol, 50 mM; 1-butanol, 32 mM; 1-pentanol, 13 mM; and 1-hexanol, 7 mM. For long-chain alcohols, 1-heptanol was less potent than 1 hexanol (estimated IC(50): 19 mM for GluR1 and 18 mM for GluR3), 1-octanol had little effect only on GluR3, and 1-nonanol and 1-decanol did not significantly inhibit both GluR1 and GluR3 responses. The observations indicate that straight chain n-alcohols exhibit a cutoff in their potency for inhibition of the function of non-NMDA glutamate receptor subunits, GluR1 and GluR3. The cutoff in potency of n-alcohols for inhibition of non-NMDA glutamate receptor function is consistent with the interpretation that alcohols affect the function of these receptor-channels by interacting with an alcohol binding site of specific dimensions on the receptor protein. PMID- 11429389 TI - Blockade by ruthenium red of tissue factor-initiated coagulation. AB - The ability of ruthenium red (RuR) to inhibit tissue factor (TF)-initiated blood coagulation was demonstrated at the protein and cellular levels as well as in human plasma. In a single-stage clotting assay, RuR concentration-dependently inhibited rabbit brain thromboplastin (rbTF)-induced coagulation and offset bacterial endotoxin (LPS)-induced monocytic TF (mTF) hypercoagulation; the IC(50)s were estimated at 7.5 and 12.3 microM, respectively. A 15-min preincubation of RuR with rbTF or monocyte suspension resulted in the pronounced inhibition with a significantly lowered IC(50) at 1.8 or 7.7 microM for rbTF or mTF procoagulation, respectively. The differences in IC(50)s between rbTF and mTF without or with the preincubation indicated that TF was a primary target for RuR action. The effect of RuR on the physiological function of TF in FVII activation was demonstrated by the proteolytic cleavage of FVII zymogen to its active forms of serine protease on Western blotting analyses. RuR readily blocked TF-catalyzed FVII activation (diminished FVIIa formation), thus down regulating the initiation of blood coagulation. Inclusion of RuR into human plasma samples in vitro significantly prolonged prothrombin time, indicating the depressed coagulation. FVII activity was inhibited by 30 - 60% depending on the dose; as a result, FX activity also decreased. However, RuR showed no effect on thrombin time. Thus, RuR inhibited FVII activation to block the initiation of coagulation. PMID- 11429390 TI - Relaxation to authentic nitric oxide and SIN-1 in rat isolated mesenteric arteries: variable role for smooth muscle hyperpolarization. AB - Authentic nitric oxide (NO; 0.1 - 10 micromoles) caused transient, dose-dependent relaxation of phenylephrine-induced tone without changing membrane potential in mesenteric arteries. Larger doses, above 10 micromoles, did not evoke more relaxation (maximal relaxation to 150 micromoles NO in denuded arteries, 69+/-7%, n=8) but stimulated muscle hyperpolarization (maximum 19+/-3 mV, n=5). The soluble guanylyl cyclase inhibitor, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; 10 microM), abolished relaxation to low doses of NO (n=4), but did not modify hyperpolarization with higher doses of NO (n=4). The potassium channel blocker charybdotoxin (ChTX; 50 nM) abolished hyperpolarization to high doses of NO and significantly reduced the maximal relaxation (to 43+/-6%, n=4; P<0.01). ODQ and ChTX together abolished tension and membrane potential change to all doses of NO (n=4). All relaxations to 3-morpholino-sydnonimine (SIN-1; 0.01 - 10 microM) were associated with hyperpolarization. When the endothelium was intact, ChTX inhibited hyperpolarization and relaxation to SIN-1 (n=5), while iberiotoxin (IbTX; 50 nM) or 4-aminopyridine (4-AP; 500 microM) reduced relaxation by 40% and 20%, respectively and by 80% in combination (n=6 in each case). In denuded arteries, relaxation to SIN-1 was unaffected by either ChTX or ODQ alone, but abolished by the inhibitors together (n=6). Alone, 4-AP did not alter relaxation, but in the presence of ODQ it reduced the maximal response by around 45% (n=6; P<0.01). 4-AP, ODQ and IbTX together inhibited relaxation to SIN-1 by 75% (n=6; P<0.01). Therefore, cyclic guanosine 3',5'-monophosphate (cyclic GMP)-independent smooth muscle hyperpolarization, possibly involving direct activation of calcium activated and voltage-sensitive potassium channels, contributes to relaxation evoked by authentic NO and SIN-1. However, the importance of each pathway depends on the source of NO and with SIN-1 the relative contribution from each pathway is modified by the endothelium. PMID- 11429391 TI - Nipecotic acid directly activates GABA(A)-like ion channels. AB - The GABA-related compound nipecotic acid is commonly used to inhibit GABA uptake. This report shows that nipecotic acid can also directly activate GABA(A)-like chloride channels. When applied to outside-out patches of paraventricular neurones, nipecotic acid (1 mM) activated inward unitary currents (approximately 3 pA at a holding potential of -60 mV, E(Cl)+44 mV). The EC(50) for ion channel activation was approximately 300 microM, 3 fold greater than that found for GABA itself in this preparation. The nipecotic acid activated channels had similar conductance and kinetic properties to those of GABA activated channels in the same patches, reversed near E(Cl) and were inhibited by bicuculline (3 microM). This study indicates that for experiments in which relatively high concentrations of nipecotic acid are used, possible direct GABA(A) receptor agonist properties should be considered. PMID- 11429392 TI - Functional characterization of alpha(1)-adrenoceptor subtypes in human skeletal muscle resistance arteries. AB - alpha(1)-adrenoceptor subtypes in human skeletal muscle resistance arteries were characterized using agonists noradrenaline (non-selective) and A61603 (alpha(1A) selective), the antagonists prazosin (non-selective), 5-methyl-urapidil (alpha(1A)-selective) and BMY7378 (alpha(1D)-selective) and the alkylating agent chloroethylclonidine (preferential for alpha(1B)). Small arteries were obtained from the non-ischaemic skeletal muscle of limbs amputated for critical limb ischaemia and isometric tension recorded using wire myography. Prazosin antagonized responses to noradrenaline with a pA(2) value of 9.18, consistent with the presence of alpha(1)-adrenoceptors, although the Schild slope (1.32) was significantly different from unity. 5-Methyl-urapidil competitively antagonized responses to noradrenaline with a pK(B) value of 8.48 and a Schild slope of 0.99, consistent with the presence of alpha(1A)-adrenoceptors. In the presence of 300 nM 5-methyl-urapidil, noradrenaline exhibited biphasic concentration response curves, indicating the presence of a minor population of a 5-methyl-urapidil resistant subtype. Contractile responses to noradrenaline were not affected by 1 microM chloroethylclonidine suggesting the absence of alpha(1B)-adrenoceptors. Maximum responses to noradrenaline and A61603 were reduced to a similar extent by 10 microM chloroethylclonidine, suggesting an effect of chloroethylclonidine at alpha(1A)-adrenoceptors at the higher concentration. BMY7378 (10 and 100 nM) had no effect on responses to noradrenaline. BMY7378 (1 microM) poorly shifted the potency of noradrenaline giving a pA(2) of 6.52. These results rule out the presence of the alpha(1D)-subtype. These results show that contractile responses to noradrenaline in human skeletal muscle resistance arteries are predominantly mediated by the alpha(1A)-adrenoceptor subtype with a minor population of an unknown alpha(1)-adrenoceptor subtype. PMID- 11429393 TI - Reversal of cardiac and renal fibrosis by pirfenidone and spironolactone in streptozotocin-diabetic rats. AB - Fibrosis leads to chronic impairment of cardiac and renal function and thus reversal of existing fibrosis may improve function and survival. This project has determined whether pirfenidone, a new antifibrotic compound, and spironolactone, an aldosterone antagonist, reverse both deposition of the major extracellular matrix proteins, collagen and fibronectin, and functional changes in the streptozotocin(STZ)-diabetic rat. Streptozotocin (65 mg kg(-1) i.v.)-treated rats given pirfenidone (5-methyl-1-phenyl-2-[1H]-pyridone; approximately 200 mg kg(-1) day(-1) as 0.2 - 2g 1(-1) drinking water) or spironolactone (50 mg kg(-1) day(-1) s.c.) for 4 weeks starting 4 weeks after STZ showed no attenuation of the increased blood glucose concentrations and increased food and water intakes which characterize diabetes in this model. STZ-treatment increased perivascular and interstitial collagen deposition in the left ventricle and kidney, and surrounding the aorta. Cardiac, renal and plasma fibronectin concentrations increased in STZ-diabetic rats. Passive diastolic stiffness increased in isolated hearts from STZ-diabetic rats. Both pirfenidone and spironolactone treatment attenuated these increases without normalizing the decreased +dP/dt(max) of STZ diabetic hearts. Left ventricular papillary muscles from STZ-treated rats showed decreased maximal positive inotropic responses to noradrenaline, EMD 57033 (calcium sensitizer) and calcium chloride; this was not reversed by pirfenidone or spironolactone treatment. STZ-treatment transiently decreased GFR and urine flow rates in isolated perfused kidneys; pirfenidone but not spironolactone prevented the return to control values. Thus, short-term pirfenidone and spironolactone treatment reversed cardiac and renal fibrosis and attenuated the increased diastolic stiffness without normalizing cardiac contractility or renal function in STZ-diabetic rats. PMID- 11429394 TI - Disturbance of the prejunctional modulation of cholinergic neurotransmission during chronic granulomatous inflammation of the mouse ileum. AB - The effect of chronic granulomatous inflammation of the intestine was studied on the prejunctional modulation of cholinergic nerve activity in the mouse ileum. Contractions to carbachol (0.01 - 0.3 microM) and to electrical field stimulation (EFS, 0.25 - 8 Hz) of enteric neurons were higher in inflamed ileum as compared to control ileum. However, when the neurally-mediated contractions to EFS were expressed as percentage of the direct smooth muscle contraction to carbachol, the responses to EFS were similar in control and inflamed ileum. Atropine (1 microM) abolished all contractions to EFS and carbachol in control and inflamed ileum. DMPP (3 - 30 microM), a nicotinic receptor agonist, induced concentration dependent contractions that were more pronounced in inflamed ileum as compared to control ileum. Hexamethonium (100 microM), a nicotinic receptor blocker, significantly inhibited the contractions to EFS in inflamed ileum but not in control ileum. In control ileum, histamine (10 - 100 microM) and the histamine H(1) receptor agonist HTMT (3 - 10 microM) inhibited the contractions to EFS concentration-dependently without affecting the contractions to carbachol. The inhibitory effect of histamine and HTMT was prevented by the histamine H(1) antagonist mepyramine (5 - 10 microM) but not by the H(2)- and H(3)-receptor antagonists cimetidine and thioperamide (both 10 microM). In chronically inflamed ileum however, histamine (10 - 100 microM) and HTMT (3 - 10 microM) failed to inhibit the contractions to EFS. The histamine H(2) and H(3) receptor agonists dimaprit and R(-)-alpha-methylhistamine did not affect the contractions to EFS in control and inflamed ileum. The alpha(2)-receptor agonist UK 14.304 (0.01 - 0.1 microM) inhibited the contractions to EFS in control and inflamed ileum without affecting the contractions to carbachol. The effect of UK 14.304 was reversed by the alpha(2)-receptor antagonist yohimbine (1 microM). The inhibitory effect of UK 14.304 on contractions to EFS was of similar potency in control and inflamed ileum. Our results suggest that the prejunctional modulation of cholinergic nerve activity by nicotinic and histaminic H(1) receptors is disturbed during chronic intestinal inflammation whereas the modulation by alpha(2)-receptors is preserved. Such a disturbance of cholinergic nerve activity may contribute to the motility disturbances that are often observed during chronic intestinal diseases in humans. PMID- 11429395 TI - The effect of the beta(2) adrenoceptor gene Thr164Ile polymorphism on human adipose tissue lipolytic function. AB - A rare beta(2)-adrenoceptor gene polymorphism, Thr164Ile, has been described that impairs receptor function when transfected into cell lines. We investigated whether the polymorphism influences native receptor function by studying lipolysis in freshly isolated subcutaneous fat cells from 236 apparently healthy subjects. Twelve subjects were heterozygous for the 164Ile variant. The fat cells of Ile carriers displayed a 6 fold increase (P=0.02) in the lipolytic EC(50) of terbutaline (a selective beta(2)-adrenoceptor agonist), but no change in the lipolytic action of dobutamine (a selective beta(1)-adrenoceptor agonist), compared with the Thr carriers. Maximum adrenoceptor agonist stimulated lipolysis did not differ between Thr and Ile carriers. The influence of two other polymorphisms (Arg16Gly and Gln27Glu) in the beta(2)-adrenoceptor gene was considered. Six 164Ile carriers also carried the 16Gly and 27Glu alleles. The latter combination occurred among 105 of the 164Thr carriers. For the 16Gly27Glu subgroup, the EC(50) of terbutaline was about 10 fold higher in 164Ile as than in 164Thr carriers (P=0.02) but there was no difference between genotypes in maximum terbutaline action. There was no difference between groups in dobutamine action. In conclusion, the 164Ile variant of the beta(2)-adrenoceptor is associated with a decreased native adipocyte receptor function, as evidenced by a marked increase in the half maximal effective concentration of the lipolytic action of a selective beta(2)-adrenoceptor agonist. This suggests that genetic variance in the beta(2)-adrenoceptor gene might be important for catecholamine function in humans, at least as far as adipocyte lipolysis is concerned. PMID- 11429396 TI - Platelet-activating factor and endotoxin activate CCAAT/enhancer binding protein in rat small intestine. AB - The transcription factor family CCAAT/enhancer binding proteins (C/EBP) is involved in inflammation via the regulation of the gene expression of various pro inflammatory cytokines and proteins. PAF and endotoxin (lipopolysaccharide, LPS) are known agents causing intestinal inflammation and injury. In this study, we examined the binding activity of C/EBP isoforms in rat small intestine in response to PAF (1.5 microg kg(-1), i.v.) or LPS (5 mg kg(-1), i.v.). We found that C/EBP is constitutively active in normal small intestine, mainly as C/EBP alpha and beta (C/EBP-beta>alpha). Both C/EBP-alpha and beta are localized in the intestinal epithelial cells: C/EBP-alpha mainly in the crypts, and C/EBP-beta in both villi and crypts, as well as in some lamina propria cells. Only minute amounts of C/EBP-delta were found. PAF rapidly upregulates the binding activity of C/EBP-alpha and beta within 30 min. The increase in C/EBP-alpha is prominent in the crypt cells, whereas the change of C/EBP-beta is more widespread. LPS also increases the binding activity of C/EBP-alpha and beta, and the response is slower than PAF. PAF synergizes with LPS to markedly activate all three subunits. The increase in C/EBP-alpha is transient, whereas the other two have a sustained elevation until 120 min. After challenge with PAF (but not LPS), small amounts of nuclear factor -kappaB (NF-kappaB) p50 and p65 subunits are found in the C/EBP DNA binding complex, indicating cross-dimerization of the two transcription families. Pretreatment of rats with pyrrolidine dithiocarbamate (PDTC) suppresses LPS-, but not PAF-, induced NF-kappaB and C/EBP binding activity, and significantly increases the C/EBP-delta subunit in LPS- or PAF-induced C/EBP complex. These results suggest that PAF and LPS activate intestinal C/EBP in vivo, probably via different pathways. PMID- 11429397 TI - Identification and quantification of phosphodiesterase 4 subtypes in CD4 and CD8 lymphocytes from healthy and asthmatic subjects. AB - In the present study, for the first time, PDE4 subtypes were identified and semi quantified in both CD4 and CD8 lymphocytes from healthy and asthmatic individuals. CD4 and CD8 lymphocytes from healthy and mild asymptomatic asthmatic subjects (receiving beta-agonist therapy only) were isolated from peripheral venous blood using appropriate antibody coated paramagnetic beads. PDE4 subtypes and beta-actin were identified by digoxigenin (DIG)-labelling reverse transcriptase-polymerase chain reaction and semi-quantified by DIG-detection enzyme-linked immunosorbance assay. In CD4 and CD8 lymphocytes PDE4A, PDE4B and PDE4D were detected, with no significant differences observed between healthy and asthmatic groups. In CD8 lymphocytes, enzyme subtype expression was lower and showed more intersubject variability. In functional studies investigating the effects of various PDE inhibitors on PHA-induced proliferation of mononuclear cells from healthy and asthmatic subjects, CDP840 (0.03 - 10 microM), rolipram (0.1 - 10 microM) and theophylline (10 microM - 1 mM) inhibited PHA-induced proliferation of mononuclear cells from healthy and asthmatic subjects in a concentration-dependent manner, although no significant difference was observed between the groups investigated. In additional studies, total monocyte cyclic AMP PDE activity was investigated in cells isolated from asthmatic subjects both prior to and 24 h after allergen challenge. Total monocyte cyclic AMP PDE activity remained unaffected following challenge of asthmatic subjects with either house dust mite or cat dander and was inhibited in a concentration dependent manner by rolipram (0.01 - 100 microM) both before and after allergen challenge. PMID- 11429398 TI - The effects of flecainide on ATP-sensitive K(+) channels in pig urethral myocytes. AB - The effects of the antiarrhythmic drug flecainide on levcromakalim-induced hyperpolarization, macroscopic and unitary K(+) currents in pig urethra were investigated using patch-clamp techniques. The effects of flecainide were also examined on currents in inside-out patches of COS7 cells expressing carboxy terminus truncated inwardly rectifying K(+) channel (Kir6.2) subunits (i.e. Kir6.2DeltaC36) which form ATP-sensitive K(+) channels (K(ATP) channels). In current-clamp mode, application of flecainide (> or =100 microM) caused a significant depolarization after the membrane potential had been hyperpolarized by levcromakalim. In voltage-clamp experiments, the levcromakalim-induced outward current was suppressed by 300 microM flecainide in quasi-physiological K(+) conditions (K(i)=51 microM). In contrast, approximately 20% of the levcromakalim induced inward current still remained even after application of 300 microM flecainide in symmetrical 140 mM K(+) conditions (K(i)=51 microM). In contrast, approximately 20% of the levcromakalim-induced inwar=126 microM). In cell attached configuration, the channel activity of the levcromakalim-induced K(ATP) channels was reversibly inhibited by flecainide (> or =30 microM) at -50 mV. Their activity was also suppressed by either disopyramide or cibenzoline. Flecainide reversibly inhibited the channel activity of Kir6.2DeltaC36 expressed in COS7 cells using inside-out configuration. Inhibitory effects of flecainide on the levcromakalim-induced currents became more potent when the value of external pH increased, although this slightly reduced the proportion of drug molecules carrying a positive charge. These results suggest that flecainide inhibits channel activity through blocking the pore site of the K(ATP) channel in pig urethra. PMID- 11429399 TI - Antioxidant activity of amiodarone on human lipoprotein oxidation. AB - Lipoprotein oxidation is crucial in atherogenic processes. Amiodarone is a lipophilic antiarrhythmic/antianginal drug which is able to influence the physicochemical status of biological lipid components. Since oxidation of lipids is affected by their physicochemical state and amiodarone binds to lipoproteins, we hypothesized that the drug may exert an antioxidant activity on human lipoprotein oxidation. Dose-dependent effects of therapeutically achievable amiodarone concentrations (1.5, 3, 5, 7 and 10 microM) were studied on copper catalysed oxidation of the non-HDL fraction in vitro. Amiodarone inhibited oxidation as judged by generation of thiobarbituric acid reactive substances (TBARS), lipid hydroperoxides (LOOH) and fluorescent products of lipoperoxidation (FPL) as well as from the kinetics of conjugated diene formation. This antioxidant activity was significant at 1.5 microM with total inhibition at 10 microM and an IC(50) of 4 microM. The primary in vivo metabolite of amiodarone, namely desethylamiodarone, also exhibited specific antioxidant properties although it was less effective than amiodarone with an IC(50) of 7 microM. In further in vivo experiments, susceptibility to copper-mediated oxidation of the non-HDL fraction was investigated before and 4 weeks after oral amiodarone administration to humans. Following treatment, significant inhibition of TBARS, LOOH and FPL generation was observed in comparison with baseline levels and a placebo-treated control group, highlighting an effective antioxidant capacity of amiodarone in vivo. Amiodarone did not change lipoprotein vitamin E and phospholipid content in vivo and did not show scavenging effects on oxidizing species involved in lipoprotein oxidation, such as peroxyl radicals, nor metal binding/inactivating properties, suggesting that physicochemical modifications of lipoprotein lipids induced by the lipophilic drug may be involved in its antioxidant activity. In conclusion, amiodarone, and its primary metabolite desethylamiodarone, show previously unrecognized antioxidant activity on human lipoprotein oxidation. This effect is also evident in vivo and at therapeutically achievable drug concentrations. Thus, amiodarone may act as an antioxidant/antiatherosclerotic agent in humans, although this issue warrants further clinical study. PMID- 11429400 TI - Effects of YM471, a nonpeptide AVP V(1A) and V(2) receptor antagonist, on human AVP receptor subtypes expressed in CHO cells and oxytocin receptors in human uterine smooth muscle cells. AB - YM471, (Z)-4'-[4,4-difluoro-5-[2-(4-dimethylaminopiperidino)-2-oxoethylidene] 2,3,4,5-tetrahydro-1H-1-benzoazepine-1-carbonyl]-2-phenylbenzanilide monohydrochloride, is a newly synthesized potent vasopressin (AVP) receptor antagonist. Its effects on binding to and signal transduction by cloned human AVP receptors (V(1A), V(1B) and V(2)) stably expressed in Chinese hamster ovary (CHO) cells, and oxytocin receptors in human uterine smooth muscle cells (USMC) were studied. YM471 potently inhibited specific [(3)H]-AVP binding to V(1A) and V(2) receptors with K(i) values of 0.62 nM and 1.19 nM, respectively. In contrast, YM471 exhibited much lower affinity for V(1B) and oxytocin receptors with K(i) values of 16.4 microM and 31.6 nM, respectively. In CHO cells expressing V(1A) receptors, YM471 potently inhibited AVP-induced intracellular Ca(2+) concentration ([Ca(2+)](i)) increase, exhibiting an IC(50) value of 0.56 nM. However, in human USMC expressing oxytocin receptors, YM471 exhibited much lower potency in inhibiting oxytocin-induced [Ca(2+)](i) increase (IC(50)=193 nM), and did not affect AVP-induced [Ca(2+)](i) increase in CHO cells expressing V(1B) receptors. Furthermore, in CHO cells expressing V(2) receptors, YM471 potently inhibited the production of cyclic AMP stimulated by AVP with an IC(50) value of 1.88 nM. In all assays, YM471 showed no agonistic activity. These results demonstrate that YM471 is a potent, nonpeptide human V(1A) and V(2) receptor antagonist which will be a valuable tool in defining the physiologic and pharmacologic actions of AVP. PMID- 11429401 TI - Human neuroglobin, a hexacoordinate hemoglobin that reversibly binds oxygen. AB - Neuroglobin is a newly discovered mammalian hemoglobin that is expressed predominately in the brain (Burmester, T., Welch, B., Reinhardt, S., and Hankeln, T. (2000) Nature 407, 520-523). Neuroglobin has less than 25% identity with other vertebrate globins and shares less than 30% identity with the annelid nerve myoglobin it most closely resembles among known hemoglobins. Spectroscopic and kinetic experiments with the recombinant protein indicate that human neuroglobin is the first example of a hexacoordinate hemoglobin in vertebrates and is similar to plant and bacterial hexacoordinate hemoglobins in several respects. The ramifications of hexacoordination and potential physiological roles are explored in light of the determination of an O(2) affinity that precludes neuroglobin from functioning in traditional O(2) storage and transport. PMID- 11429402 TI - Defective cytochrome c-dependent caspase activation in ovarian cancer cell lines due to diminished or absent apoptotic protease activating factor-1 activity. AB - Apoptosis via the mitochondrial pathway requires release of cytochrome c into the cytosol to initiate formation of an oligomeric apoptotic protease-activating factor-1 (APAF-1) apoptosome. The apoptosome recruits and activates caspase-9, which in turn activates caspase-3 and -7, which then kill the cell by proteolysis. Because inactivation of this pathway may promote oncogenesis, we examined 10 ovarian cancer cell lines for resistance to cytochrome c-dependent caspase activation using a cell-free system. Strikingly, we found that cytosolic extracts from all cell lines had diminished cytochrome c-dependent caspase activation compared with normal ovarian epithelium extracts. The resistant cell lines expressed APAF-1 and caspase-9, -3, and -7; however, each demonstrated diminished APAF-1 activity relative to the normal ovarian epithelium cell lines. A competitive APAF-1 inhibitor may account for the diminished APAF-1 activity because we did not detect dominant APAF-1 inhibitors, altered APAF-1 isoform expression, or APAF-1 deletion, degradation, or mutation. Lack of APAF-1 activity correlated in some but not all cell lines with resistance to apoptosis. These data suggest that regulation of APAF-1 activity may be important for apoptosis regulation in some ovarian cancers. PMID- 11429403 TI - Protein dynamics enhance electronic coupling in electron transfer complexes. AB - Electron-transferring flavoproteins (ETFs) from human and Paracoccus denitrificans have been analyzed by small angle x-ray scattering, showing that neither molecule exists in a rigid conformation in solution. Both ETFs sample a range of conformations corresponding to a large rotation of domain II with respect to domains I and III. A model of the human ETF.medium chain acyl-CoA dehydrogenase complex, consistent with x-ray scattering data, indicates that optimal electron transfer requires domain II of ETF to rotate by approximately 30 to 50 degrees toward domain I relative to its position in the x-ray structure. Domain motion establishes a new "robust engineering principle" for electron transfer complexes, tolerating multiple configurations of the complex while retaining efficient electron transfer. PMID- 11429404 TI - Two vicinal cysteines confer a peculiar redox regulation to low molecular weight protein tyrosine phosphatase in response to platelet-derived growth factor receptor stimulation. AB - Low molecular weight protein tyrosine phosphatase (LMW-PTP) is an enzyme involved in platelet-derived growth factor (PDGF)-induced mitogenesis and cytoskeleton rearrangement because it is able to bind and dephosphorylate the activated receptor. LMW-PTP presents two cysteines in positions 12 and 17, both belonging to the catalytic pocket; this is a unique feature of LMW-PTP among all protein tyrosine phosphatases. Our previous results demonstrated that in vitro LMW-PTP is oxidized by either H(2)O(2) or nitric oxide with the formation of a disulfide bond between Cys-12 and Cys-17. This oxidation leads to reversible enzyme inactivation because treatment with reductants permits catalytic activity rescue. In the present study we investigated the in vivo inactivation of LMW-PTP by either extracellularly or intracellularly generated H(2)O(2), evaluating its action directly on its natural substrate, PDGF receptor. LMW-PTP is oxidized and inactivated by exogenous oxidative stress and recovers its activity after oxidant removal. LMW-PTP is oxidized also during PDGF signaling, very likely upon PDGF induced H(2)O(2) production, and recovers its activity within 40 min. Our results strongly suggest that reversibility of in vivo LMW-PTP oxidation is glutathione dependent. In addition, we propose an intriguing and peculiar role of Cys-17 in the formation of a S-S intramolecular bond, which protects the catalytic Cys-12 from further and irreversible oxidation. On the basis of our results we propose that the presence of an additional cysteine near the catalytic cysteine could confer to LMW-PTP the ability to rapidly recover its activity and finely regulate PDGF receptor activation during both extracellularly and intracellularly generated oxidative stress. PMID- 11429405 TI - The POU domain factor Skin-1a represses the keratin 14 promoter independent of DNA binding. A possible role for interactions between Skn-1a and CREB-binding protein/p300. AB - The genes encoding keratin 5 and 14 are highly expressed in the basal cell layer keratinocytes of the epidermis, but both genes are silenced when keratinocytes move into the suprabasal compartment. The POU homeodomain factors Skn-1a and Tst 1, which are expressed in epidermis, may play a role in the suprabasal repression of the keratin 5 and 14 genes because keratin 14 mRNA expression persists in suprabasal cells in Skn-1/Tst-1 double knockout mice. In transfection experiments, both Skn-1a and Tst-1 repress the keratin 14 promoter, with the POU domain being sufficient for repression. The region of the keratin 14 gene sufficient and required for repression by Skn-1a is a 100-base pair sequence lacking POU-binding sites adjacent to the transcription start site. DNA-binding defective mutants of Skn-1a and Tst-1 are as effective at mediating repression as the wild type proteins, suggesting that protein-protein interactions rather than direct DNA binding are important for repression. We also show that CREB-binding protein (CBP)/p300 co-activators are strong activators of keratin 14 gene expression, acting through sequences close to the keratin 14 promoter. Further, CBP interacts directly with the POU domain of Skn-1a, and increasing concentrations of CBP can overcome Skn-1a-mediated repression, suggesting that POU domain factors may repress keratin 14 gene expression by interfering with the activity of co-activators such as CBP/p300. PMID- 11429406 TI - Regulation of protein-tyrosine phosphatases alpha and epsilon by calpain-mediated proteolytic cleavage. AB - The precise subcellular localization of non-receptor tyrosine phosphatases is a major factor in regulating their physiological functions. We have previously shown that cellular processing of protein-tyrosine phosphatase epsilon (PTP epsilon) generates a physiologically distinct, cytoplasmic form of this protein, p65 PTP epsilon. Here we describe a novel protein form of the related receptor type tyrosine phosphatase alpha (RPTP alpha), p66 PTP alpha, which is detected in nearly all cell types where RPTP alpha is expressed. Both p66 PTP alpha and p65 PTP epsilon are produced by calpain-mediated proteolytic cleavage in vivo. Cleavage is inhibited in living cells by a variety of calpain inhibitors, can be induced in primary cortical neurons treated with calcium chloride, and is observed in lysates of brain or of cultured cells following addition of purified calpain. Cleavage occurs within the intracellular juxtamembrane domain of RPTP alpha, releasing the phosphatase catalytic domains from their membranal anchors and translocating them to the cytoplasm. Translocation reduces the ability of PTPalpha to act on membrane-associated substrates, as it loses its ability to dephosphorylate Src at its C-terminal regulatory site, and its ability to dephosphorylate the Kv2.1 voltage-gated potassium channel is severely impaired. In all, the data indicate that control of phosphatase function via post translational processing occurs also among receptor-type phosphatases, and demonstrate the molecular complexity of regulating these parameters within the PTP alpha/PTP epsilon phosphatase subfamily. PMID- 11429407 TI - Cross-linking of human multidrug resistance P-glycoprotein by the substrate, tris (2-maleimidoethyl)amine, is altered by ATP hydrolysis. Evidence for rotation of a transmembrane helix. AB - We identified a thiol-reactive substrate, Tris-(2-maleimidoethyl)amine (TMEA), to explore the contribution of the TM segments 6 and 12 of the human multidrug resistance P-glycoprotein (P-gp) during transport. TMEA is a trifunctional maleimide and stimulated the ATPase activity of Cys-less P-gp about 7-fold. Cysteine-scanning mutagenesis of TM12 showed that the activity of mutant V982C was inhibited by TMEA. P-gp mutants containing V982C (TM12) and another cysteine in TM6 were constructed and tested for cross-linking with TMEA. A cross-linked product was observed in SDS-polyacrylamide gel electrophoresis for mutant L339C(TM6)/V982C(TM12). Cross-linking by TMEA also inhibited the ATPase activity of the mutant protein. Substrates such as cyclosporin A, vinblastine, colchicine, or verapamil inhibited cross-linking by TMEA. In the presence of ATP at 37 degrees C, cross-linking of mutant L339C/V982C was decreased. In contrast, there was enhanced cross-linking of mutant F343C(TM6)/V982C(TM12) in the presence of ATP. These results show that cross-linking must be within the drug-binding domain, that residues L339C(TM6)/V982C(TM12) must be at least 10 A apart, and that ATP hydrolysis promotes rotation of one or both TM helices. PMID- 11429408 TI - CYP98A3 from Arabidopsis thaliana is a 3'-hydroxylase of phenolic esters, a missing link in the phenylpropanoid pathway. AB - The 4- and 5-hydroxylations of phenolic compounds in plants are catalyzed by cytochrome P450 enzymes. The 3-hydroxylation step leading to the formation of caffeic acid from p-coumaric acid remained elusive, however, alternatively described as a phenol oxidase, a dioxygenase, or a P450 enzyme, with no decisive evidence for the involvement of any in the reaction in planta. In this study, we show that the gene encoding CYP98A3, which was the best possible P450 candidate for a 3-hydroxylase in the Arabidopsis genome, is highly expressed in inflorescence stems and wounded tissues. Recombinant CYP98A3 expressed in yeast did not metabolize free p-coumaric acid or its glucose or CoA esters, p coumaraldehyde, or p-coumaryl alcohol, but very actively converted the 5-O shikimate and 5-O-d-quinate esters of trans-p-coumaric acid into the corresponding caffeic acid conjugates. The shikimate ester was converted four times faster than the quinate derivative. Antibodies directed against recombinant CYP98A3 specifically revealed differentiating vascular tissues in stem and root. Taken together, these data show that CYP98A3 catalyzes the synthesis of chlorogenic acid and very likely also the 3-hydroxylation of lignin monomers. This hydroxylation occurs on depsides, the function of which was so far not understood, revealing an additional and unexpected level of networking in lignin biosynthesis. PMID- 11429409 TI - DNA binding-independent transcriptional activation by the androgen receptor through triggering of coactivators. AB - Androgens have critical roles in the development and maintenance of the male reproductive system and are important for progression of prostate cancer. The effects of androgens are mediated by the androgen receptor (AR), which is a ligand-modulated transcription factor that belongs to the nuclear receptor superfamily. In the presence of androgens, AR binds to androgen response elements in the vicinity of androgen receptor target genes and activates transcription. In addition, liganded AR can interfere with the activity of other transcription factors, such as activator protein-1 and nuclear factor kappaB, for which DNA binding by AR is not necessary. In this study, we describe a novel ligand dependent transactivation function for AR that is independent of its DNA binding ability. AR dramatically increased the intrinsic transcriptional activity of the nuclear receptor coactivators glucocorticoid receptor-interacting protein-1 (GRIP1), cAMP response element-binding protein-binding protein, and p300 that are tethered to DNA. This "triggering" phenomenon required both similar and distinctly different regions of AR compared with those needed for ligand dependent transactivation from androgen-responsive elements. Furthermore, the domains of GRIP1 required for triggering by AR are different from those required when GRIP1 serves as a coactivator for AR at androgen-responsive promoters. These data suggest that triggering may constitute an important part of the mechanism by which AR regulates transcription. PMID- 11429410 TI - The digalactosyldiacylglycerol (DGDG) synthase DGD1 is inserted into the outer envelope membrane of chloroplasts in a manner independent of the general import pathway and does not depend on direct interaction with monogalactosyldiacylglycerol synthase for DGDG biosynthesis. AB - Galactolipids make up the bulk of chloroplast lipids. Therefore, the genes involved in the synthesis of the galactolipids monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) play a critical role in chloroplast development. In this study, we analyzed the subcellular localization of the Arabidopsis DGDG synthase DGD1, which was recently identified by complementation of the Arabidopsis dgd1 mutant. In vitro import experiments demonstrated that DGD1 was targeted to the chloroplast outer envelope in an ATP-independent manner. DGD1 could not be extracted from the membranes by high salt or alkali, suggesting that it is an integral membrane protein. Uptake experiments with truncated versions of DGD1 indicated that the information for targeting and insertion into the outer envelope resides in the N-terminal half of DGD1, but not in the first 33 amino acids. DGD1 apparently does not contain a cleavable signal peptide. Antibodies to Arabidopsis DGD1 detected a 90-kDa protein localized to the chloroplast envelopes of both pea and Arabidopsis. Transformation of DGD1 constructs into cyanobacteria resulted in the expression of active DGDG synthase and demonstrated that DGDG synthesis depends on MGDG lipid, but does not require direct interaction with the plant MGDG synthase. PMID- 11429411 TI - Identification of a nonconserved amino acid residue in multidrug resistance protein 1 important for determining substrate specificity: evidence for functional interaction between transmembrane helices 14 and 17. AB - Murine multidrug resistance protein 1 (mrp1), differs from its human ortholog (MRP1) in that it fails to confer anthracycline resistance and transports the MRP1 substrate, 17beta-estradiol 17-(beta-d-glucuronide) (E(2)17betaG), very poorly. By mutating variant residues in mrp1 to those present in MRP1, we identified Glu(1089) of MRP1 as being critical for anthracycline resistance. However, Glu(1089) mutations had no effect on E(2)17betaG transport. We have now identified a nonconserved amino acid within the highly conserved COOH-proximal transmembrane helix of MRP1/mrp1 that is important for transport of the conjugated estrogen. Converting Ala(1239) in mrp1 to Thr, as in the corresponding position (1242) in MRP1, increased E(2)17betaG transport 3-fold. Any mutation of mrp1 Ala(1239), including substitution with Thr, decreased resistance to vincristine and VP-16 without altering anthracycline resistance. However, introduction of a second murine to human mutation, Q1086E, which alone selectively increases anthracycline resistance, into mrp1A1239T restored resistance to both vincristine and VP-16. To confirm the importance of MRP1 Thr(1242) for E(2)17betaG transport and drug resistance, we mutated this residue to Ala, Cys, Ser, Leu, and Lys. These mutations decreased E(2)17betaG transport 2 fold. Conversion to Asp eliminated transport of the estrogen conjugate and also decreased leukotriene C(4) transport approximately 2-fold. The mutations also reduced the ability of MRP1 to confer resistance to all drugs tested. As with mrp1, introduction of a second mutation based on the murine sequence to create MRP1E1089Q/T1242A restored resistance to vincristine and VP-16, but not anthracyclines, without affecting transport of leukotriene C(4) and E(2)17betaG. These results demonstrate the important role of Thr(1242) for E(2)17betaG transport. They also reveal a highly specific functional relationship between nonconserved amino acids in TM helices 14 and 17 of both mrp1 and MRP1 that enables both proteins to confer similar levels of resistance to vincristine and VP-16. PMID- 11429412 TI - Activation of estrogen receptor blocks interleukin-6-inducible cell growth of human multiple myeloma involving molecular cross-talk between estrogen receptor and STAT3 mediated by co-regulator PIAS3. AB - Estrogen receptors (ERs)(1) highly expressed by multiple myeloma (MM) cells and stimulation of estrogenic ligands leads to cell apoptosis. Interleukin (IL)-6 is a major growth factor in the pathogenesis of MM. However, little is known concerning the molecular consequences of ER activation on IL-6-regulated MM cell growth. Here we show that the ER agonist 17 beta-estradiol completely abolished IL-6-inducible MM cell proliferation. By contrast, the ER antagonist ICI 182,780 overcame the inhibitory effect of estrogen. Estrogen blocked STAT3 DNA binding and transactivation but failed to affect the mRNA expression of IL-6 receptor chains or activation of JAK2 and STAT3. Estrogen-activated ER did not associate directly with STAT3. Estrogen induced the mRNA expression of PIAS3 (protein inhibitor of activated STAT3) and increased PIAS3 physical association with STAT3, suggesting a possible mechanism of STAT3 inhibition requiring PIAS3 as a co-regulator modulating the cross-talk between ER and STAT3. These data directly demonstrate STAT3 to be a molecular participant in ER inhibition of the IL-6 signaling pathway in human MM cells and provides the molecular basis for the potential use of estrogenic ligands in the treatment of MM or other tumors where IL-6 has an autocrine or paracrine role. PMID- 11429413 TI - Structure and activity of the insect cytokine growth-blocking peptide. Essential regions for mitogenic and hemocyte-stimulating activities are separate. AB - Growth-blocking peptide (GBP) is a 25-amino acid insect cytokine found in Lepidopteran insects that possesses diverse biological activities such as larval growth regulation, cell proliferation, and stimulation of immune cells (plasmatocytes). The tertiary structure of GBP consists of a structured core that contains a disulfide bridge and a short antiparallel beta-sheet (Tyr(11)-Arg(13) and Cys(19)-Pro(21)) and flexible N and C termini (Glu(1)-Gly(6) and Phe(23) Gln(25)). In this study, deletion and point mutation analogs of GBP were synthesized to investigate the relationship between the structure of GBP and its mitogenic and plasmatocyte spreading activity. The results indicated that deletion of the N-terminal residue, Glu(1), eliminated all plasmatocyte spreading activity but did not reduce mitogenic activity. In contrast, deletion of Phe(23) along with the remainder of the C terminus destroyed all mitogenic activity but only slightly reduced plasmatocyte spreading activity. Therefore, the minimal structure of GBP containing mitogenic activity is 2-23 GBP, whereas that with plasmatocyte spreading activity is 1-22 GBP. NMR analysis indicated that these N- and C-terminal deletion mutants retained a similar core structure to wild-type GBP. Replacement of Asp(16) with either a Glu, Leu, or Asn residue similarly did not alter the core structure of GBP. However, these mutants had no mitogenic activity, although they retained about 50% of their plasmatocyte spreading activity. We conclude that specific residues in the unstructured and structured domains of GBP differentially affect the biological activities of GBP, which suggests the possibility that multifunctional properties of this peptide may be mediated by different forms of a GBP receptor. PMID- 11429414 TI - Induction of cellular antioxidative stress genes through heterodimeric transcription factor Nrf2/small Maf by antirheumatic gold(I) compounds. AB - Gold(I)-containing compounds have long been used in the treatment of rheumatoid arthritis (RA), but the molecular mechanism of their action has remained largely unknown. In this paper we have demonstrated that gold(I) drugs selectively activate the DNA binding of a heterodimer consisting of the basic-leucine zipper transcription factors Nrf2 and small Maf. Once bound to its recognition DNA sequence termed antioxidant-responsive element or Maf-recognition element, Nrf2/small Maf induces a set of antioxidative stress genes, including heme oxygenase-1 and gamma-glutamylcysteine synthetase, whose products have been demonstrated to contribute to the scavenging of reactive oxygen species and to exhibit anti-inflammatory effects. Our findings suggest that stimulation of antioxidative stress response through activation of Nrf2/small Maf may be a pharmacologically important part of the actions of gold(I) drugs for the treatment of rheumatoid arthritis. Alternatively, activation of Nrf2/small Maf may be a protective response of cells against toxic effects of the drugs. PMID- 11429415 TI - A single point mutation at the 3'-untranslated region of Ran mRNA leads to profound changes in lipopolysaccharide endotoxin-mediated responses. AB - By functional cDNA expression cloning, we have previously established that Ran is important in lipopolysaccharide (LPS) signaling. This was achieved by functional comparison between two cDNAs, differing by a single base substitution within the 3'-untranslated region of the cDNA. This point mutation results in a striking RNA conformational change. No dramatic difference in total RNA at steady state could be found between the two molecules. However, at the protein level, RanC/d (from 870C mRNA) was 5-10-fold higher than RanT/n (from 870T mRNA) and this difference was not observed in non-hematopoietic cells transduced with the same vectors. This tissue-specific difference correlated with a difference in LPS endotoxin responses in corresponding hematopoietic cells. Importantly, the amounts of Ran- C/d and RanT/n proteins were similar initially but the difference became obvious with time. Both Ran proteins migrated from the cytoplasm to the nucleus, but Ran from RanC/d migrated faster than that of RanT/n. RanT/n protein preferentially remained in the cytoplasm and its overall amount was reduced at steady state, consistent with its degradation by intracellular proteases known to be involved in LPS-mediated signal transduction. As the two proteins are identical, the faster RanC/d nuclear localization and a preferred initial cytoplasmic RanT/n distribution suggest a difference in mRNA intracellular localization between the two molecules, as dictated by their RNA structural difference. By pulse-chase experiments, RanC/d proteins are more resistant to degradation than RanT/n protein; there also appear to have two populations of RanT/n proteins, one may reside in the cytoplasm and the other, in the nucleus. More RanC/d GTPase accumulated in the nuclei would conceivably alter the potency of signal transduction and therefore down-modulate LPS-mediated biological responses. PMID- 11429416 TI - Mutation of residues 423 (Met/Ile), 444 (Thr/Met), and 506 (Asn/Ser) confer cholesteryl esterase activity on rat lung carboxylesterase. Ser-506 is required for activation by cAMP-dependent protein kinase. AB - Site-directed mutagenesis is used to identify amino acid residues that dictate reported differences in substrate specificity between rat hepatic neutral cytosolic cholesteryl ester hydrolase (hncCEH) and rat lung carboxylesterase (LCE), proteins differing by only 4 residues in their primary sequences. Beginning with LCE, the substitution Met(423) --> Ile(423) alone or in combination with other mutations increased activity with p-nitrophenylcaprylate (PNPC) relative to more hydrophilic p-nitrophenylacetate (PNPA), typical of hncCEH. The substitution Thr(444) --> Met(444) was necessary but not sufficient for expression of cholesteryl esterase activity in COS-7 cells. The substitution Asn(506) --> Ser(506), creating a potential phosphorylation site, uniformly increased activity with both PNPA and PNPC, was necessary but not sufficient for expression of cholesteryl esterase activity and conferred susceptibility to activation by cAMP-dependent protein kinase, a property of hncCEH. The 3 mutations in combination were necessary and sufficient for expression of cholesteryl esterase activity by the mutated LCE. The substitution Gln(186) --> Arg(186) selectively reduced esterase activity with PNPA and PNPC but was not required for cholesteryl esterase activity. Homology modeling from x-ray structures of acetylcholinesterases is used to propose three-dimensional models for hncCEH and LCE that provide insight into the effects of these mutations on substrate specificity. PMID- 11429417 TI - The Journal of Clinical Pathology online (http://www.jclinpath.com): free for the developing countries. PMID- 11429418 TI - Histological typing of lung and pleural tumours: third edition. PMID- 11429419 TI - ACP Best Practice No 165: front line tests for the investigation of suspected porphyria. AB - The porphyrias are uncommon disorders of haem biosynthesis and their effective management requires prompt and accurate diagnosis. This article describes methods for the determination of urinary porphobilinogen, urinary and faecal total porphyrins, and total porphyrins in erythrocytes and plasma that are suitable for use in non-specialist laboratories. The selection and interpretation of these methods, and the indications for further, more specialised, investigation are discussed. PMID- 11429420 TI - UK NEQAS for leucocyte immunophenotyping: the first 10 years. AB - In the past decade, cellular immunophenotyping has become a new discipline in diagnostic haematology and immunology, and is invaluable in the rapid diagnosis of leukaemia and monitoring disease progression in human immunodeficiency virus infected individuals. The introduction of bench top flow cytometers has meant that immunophenotyping is now also used for the quantitation of CD34(+) peripheral blood stem cells (PBSCs) to ensure the correct timing and adequacy of haematopoietic progenitor cell harvests. Furthermore, flow cytometry has become an important tool for the counting of leucocytes in blood components after leucocyte depletion. Because this new discipline is now such a major diagnostic and prognostic tool in the clinical arena, its use must be subject to both internal and external quality control. Such a requirement was first recognised as early as 1986 when an Inter-Regional Quality Assessment Scheme (IRQAS) was initiated for laboratories that undertook the immunocytochemical diagnosis of leukaemia using the alkaline phosphates anti-alkaline phosphatase technique. This programme began with around 25 UK laboratories. In 1990, after the introduction of two more programmes (one for leukaemia diagnosis using UV microscopy and latterly flow cytometry, and one for the enumeration of CD4(+) T cells) the IRQAS achieved UK National External Quality Assessment Scheme (UK NEQAS) status and changed its title to UK NEQAS for Leucocyte Immunophenotyping. In the past decade the once small IRQAS programme has evolved into the largest international scheme of its kind, providing EQA to over 650 laboratories world wide for leukaemia immunophenotyping, lymphocyte subset analysis, PBSCs, and more recently low level leucocyte counting. Over the years, this EQA programme has highlighted important problems, such as the inappropriate use of fluorochromes and antibody titre, and the identification of effective gating strategies, all of which have contributed directly to the high interlaboratory variations seen in cellular immunophenotyping. Furthermore, particularly in absolute counting of lymphocyte subsets, PBSCs, and the enumeration of low numbers of leucocytes, UK NEQAS for Leucocyte Immunophenotyping programmes have been instrumental in highlighting the differences that occur between single and dual platform flow cytometric technologies. As a result of these findings, UK NEQAS for Leucocyte Immunophenotyping has helped to reduce the variation seen on an interlaboratory basis and enabled greater standardisation both in the UK and internationally. These advances have been attributable to the development, by UK NEQAS for Leucocyte Immunophenotyping, of a unique whole blood stabilising process that ensures the retention of the physical characteristics (both light scatter and antigenic profile) required of cells to ensure successful cellular immunophenotyping. This major technological advancement has enabled the distribution of specimens for EQA purposes on a global scale that have minimal matrix effect and behave in a manner identical to fresh blood for several months after stabilisation. PMID- 11429421 TI - Ataxia telangiectasia gene mutations in leukaemia and lymphoma. AB - Ataxia telangiectasia (AT) is a rare multisystem, autosomal, recessive disease characterised by neuronal degeneration, genome instability, and an increased risk of cancer. Approximately 10% of AT homozygotes develop cancer, mostly of the lymphoid system. Lymphoid malignancies in patients with AT are of both B cell and T cell origin, and include Hodgkin's lymphoma, non-Hodgkin's lymphoma, and several forms of leukaemia. The AT locus was mapped to the chromosomal region 11q22-23 using genetic linkage analysis in the late 1980s and the causative gene was identified by positional cloning several years later. The ATM gene encodes a large protein that belongs to a family of kinases possessing a highly conserved C terminal kinase domain related to the phosphatidylinositol 3-kinase domain. Members of this kinase family have been shown to function in DNA repair and cell cycle checkpoint control following DNA damage. Recent studies indicate that ATM is activated primarily in response to double strand breaks and may be considered a caretaker of the genome. Most mutations in ATM result in truncation and destabilisation of the protein, but certain missense and splicing errors have been shown to produce a less severe phenotype. AT heterozygotes have a slightly increased risk of breast cancer. Atm deficient mice exhibit many of the symptoms found in patients with AT and have a high frequency of thymic lymphoma. The association between mutation of the ATM gene and a high incidence of lymphoid malignancy in patients with AT, together with the development of lymphoma in Atm deficient mice, supports the proposal that inactivation of the ATM gene may be of importance in the pathogenesis of sporadic lymphoid malignancy. Loss of heterozygosity at 11q22-23 (the location of the ATM gene) is a common event in lymphoid malignancy. Frequent inactivating mutations of the ATM gene have been reported in patients with rare sporadic T cell prolymphocytic leukaemia (T-PLL), B cell chronic lymphocytic leukaemia (B-CLL), and most recently, mantle cell lymphoma (MCL). In contrast to the ATM mutation pattern in AT, the most frequent nucleotide changes in these sporadic lymphoid malignancies were missense mutations. The presence of inactivating mutations, together with the deletion of the normal copy of the ATM gene in some patients with T-PLL, B-CLL, and MCL, establishes somatic inactivation of the ATM gene in the pathogenesis of lymphoid malignancies, and strongly suggests that ATM functions as a tumour suppressor. The presence of missense mutations in the germline of patients with B-CLL has been reported, suggesting that some patients with B-CLL may be constitutional AT heterozygotes. The putative hereditary predisposition of B-CLL, although intriguing, warrants further investigation. PMID- 11429422 TI - Paraffin wax embedded muscle is suitable for the diagnosis of muscular dystrophy. AB - AIM: At present, the diagnosis of muscular dystrophy is made by means of immunohistochemistry on frozen sections. The aim of this study was to develop a sensitive and reproducible immunohistochemical method for use on formalin fixed, paraffin wax embedded sections for the demonstration of dystrophin associated proteins and other muscle associated antigens. METHODS: All the cases studied were from the files of the department of histopathology, Great Ormond Street Hospital for Children NHS Trust. Immunohistochemistry was performed on paraffin wax embedded sections with heat mediated antigen retrieval and overnight incubation with the antibodies at room temperature. Four different pretreatment buffers were tested in the attempt to optimise the immunostaining. Frozen sections were run in parallel for direct comparison. RESULTS: All the antibodies except delta sarcoglycan gave strong, consistent immunostaining in paraffin wax embedded sections, comparable with the frozen sections. The most consistent results were obtained using citrate/EDTA as the pretreatment buffer. CONCLUSION: A reliable and reproducible technique has been established, using a heat mediated citrate/EDTA buffer antigen retrieval method, which works well for most of the antibodies needed to make the diagnosis of muscular dystrophy in formalin fixed, paraffin wax embedded sections. This technique overcomes some of the inherent problems encountered using frozen muscle tissue and it could become a valuable tool for the diagnosis of muscular dystrophy. PMID- 11429423 TI - The histopathology of non-steroidal anti-inflammatory drug induced gastroduodenal damage: correlation with Helicobacter pylori, ulcers, and haemorrhagic events. AB - AIMS: The spectrum of microscopic lesions resulting from the chronic use of non steroidal anti-inflammatory drugs (NSAIDs), known as chemical gastritis, remains unclear, and the variable prevalence reported in different studies makes this issue a matter of lively debate. The aim of this study was to evaluate the prevalence and importance of chemical gastritis in patients regularly taking NSAIDs. Owing to the high prevalence of Helicobacter pylori infection, particularly in subjects over 60 years of age, and in view of a possible association with damage, the presence of H pylori infection in the same tissue sample was also determined in all patients. METHODS: One hundred and ninety seven subjects were enrolled, 118 of whom were receiving chronic treatment with NSAIDs and 79 of whom were controls, pair matched for age, sex, and clinical symptoms (ulcer-like dyspepsia or upper digestive tract haemorrhage). Antral biopsies taken during upper gastroduodenal endoscopy were assessed for chemical gastritis according to a modified version of Dixon's score, and for Helicobacter correlated chronic active gastritis, according to the updated Sydney system. RESULTS: Chemical gastritis was identified in 11 patients taking NSAIDs (9%) and in four controls (5%) (p < 0.05). Helicobacter pylori was detected in 53 patients taking NSAIDs (45%) and in 34 controls (43%). Patients taking NSAIDs had a significantly higher number of erosions and ulcers and worse endoscores than controls. The presence of H pylori did not appear to increase histological damage, ulcer prevalence, or haemorrhagic events. CONCLUSIONS: Chemical gastritis is present in a limited number of patients regularly taking NSAIDs, and is not strongly correlated with NSAID induced damage. In many cases of peptic ulcer or upper gastrointestinal bleeding in patients taking NSAIDs, the presence of chemical gastritis or H pylori infection cannot solely account for the development of mucosal damage. PMID- 11429424 TI - Microsatellite instability in inflamed and neoplastic epithelium in ulcerative colitis. AB - BACKGROUND: Several genetic alterations have been documented in dysplasia and cancer developing in ulcerative colitis (UC). However, the microsatellite instability (MSI) status has rarely been described, especially in the inflamed epithelium of UC. AIMS: To study MSI status during neoplastic and inflammatory changes in UC. METHODS: Seventy five surgically resected samples of colorectal mucosa, taken from 16 colectomy specimens of patients with UC were examined: five patients had a long duration with dysplasia or cancer (UC-LD with neoplasm), seven patients had a long duration without neoplastic changes (UC-LD without neoplasm), and four patients had a short duration without neoplastic changes (UC SD). In addition to MSI status examined by six microsatellite markers, p53 expression was compared among the three groups. RESULTS: With regard to non neoplastic inflamed epithelium, MSI in two or more loci (MSI> or =2) was seen more frequently in the UC-LD without neoplasm group than in the UC-SD group (six of 14 v one of 12; p = 0.060), and significantly more often than in the UC-LD with neoplasm group (six of 14 v two of 23; p = 0.016). In the UC-LD without neoplasm group, MSI> or =2 was detected significantly more frequently in patients with severe inflammation than in those with mild inflammation (six of nine v none of five; p = 0.028). With regard to neoplastic epithelium in the UC-LD with neoplasm group, MSI in two or more loci was found in three of 17, and p53 overexpression was seen in 11 of 17 of the neoplastic lesions. CONCLUSIONS: A high incidence of MSI in long standing UC with severe inflammation probably reflects genomic instability caused by repeated inflammatory stress. Thus, the influence of inflammation should be considered when estimating MSI in UC. It is possible that changes in p53 expression are important in the development of cancer in UC. PMID- 11429425 TI - Expression of transcription factor AP-2 in colorectal adenomas and adenocarcinomas; comparison of immunohistochemistry and in situ hybridisation. AB - AIMS: To investigate whether the three different AP-2 isoforms are expressed differently in colorectal adenomas and carcinomas. METHODS: The study comprised 43 randomly selected patients diagnosed and treated at Kuopio University Hospital in 1996 for colorectal adenocarcinoma (n = 30) and colorectal adenoma (n = 13). The expression of AP-2alpha, AP-2beta, and AP-2gamma was analysed by immunohistochemistry (IHC) and the mRNA status of AP-2alpha was determined by in situ hybridisation (ISH) and confirmed by reverse transcription polymerase chain reaction (RT-PCR). AP-2 expression patterns were correlated with clinicopathological variables. RESULTS: In adenomas and carcinomas, AP-2beta cytoplasmic positivity was higher than that of AP-2alpha or AP-2gamma. AP-2alpha expression was reduced in advanced Dukes's stage carcinomas. In high grade carcinomas, both AP-2alpha and AP-2gamma expression was reduced. ISH demonstrated increased AP-2alpha values in high grade carcinomas. Seven of 30 carcinoma specimens displayed a moderate or strong mRNA signal, despite being negative for AP-2alpha protein. RT-PCR from AP-2alpha mRNA and protein positive tumours confirmed that the positive signal in ISH originated from the exon 2 of TFAP2A. CONCLUSIONS: AP-2alpha was reduced in advanced Dukes's stage adenocarcinomas. Together with reduced AP-2gamma expression in high grade carcinomas, this might contribute to tumour progression. The discrepancy between mRNA and protein expression suggests that post-transcriptional regulatory mechanisms might modify the availability of functional AP-2alpha protein in colorectal carcinoma. PMID- 11429426 TI - p53 codon 72 polymorphism and human papillomavirus associated skin cancer. AB - BACKGROUND/AIMS: Non-melanoma skin cancers frequently harbour multiple human papillomavirus (HPV) types. A recent report suggests that a polymorphism of the p53 tumour suppressor gene that results in the substitution of a proline residue with an arginine residue at position 72 of the p53 protein might act as a risk factor in HPV associated malignancies. This study aimed to determine the following: (1) the relation between HPV infection and the development of cutaneous squamous cell carcinoma (SCC), and (2) whether there is a correlation between p53 codon 72 polymorphism and the development of SCC. METHODS: Blood samples were taken from 55 patients with skin cancer (both renal transplant recipients and immunocompetent patients with skin cancer) and 115 ethnically matched volunteers. A polymerase chain reaction based assay was used to determine p53 codon 72 genotypes. In addition, 49 benign and malignant lesions from 34 of the patients with skin cancer and 20 normal human skin samples from 20 of the control volunteers were examined for HPV. RESULTS: The proportions of p53 codon 72 genotypes found were 78% arginine homozygous, 2% proline homozygous, and 20% heterozygous among patients with skin cancer and 79% arginine homozygous, 3.5% proline homozygous, and 17.5% heterozygous among the control population. Statistical analysis showed no significant differences in the distribution of the two p53 isoforms between the patients with skin cancer and the control population. The predominant viral types detected in both the patients and the control group were EV associated HPVs, although the incidence was lower in normal skin samples than in malignant lesions or viral warts. CONCLUSIONS: These results suggest that in a Celtic population there is no correlation between the presence of HPV, the p53 codon 72 arginine polymorphism, and the development of skin cancer. PMID- 11429427 TI - Prognostic comparative study of S-phase fraction and Ki-67 index in breast carcinoma. AB - AIMS: To investigate the prognostic value of recently proposed flow cytometric S phase fraction (SPF) variables (average SPF and SPF tertiles) compared with conventional SPF, and to compare the one with the best predictive value with the immunohistochemical Ki-67 index in breast carcinoma. METHODS: A short term follow up study (median, 39.6 months) of a large series of patients (n = 306) was conducted. DNA ploidy was analysed on fresh/frozen tumour samples by flow cytometry, and the SPF was calculated from the DNA histogram using an algorithm. The Ki-67 index was assessed on paraffin wax embedded material by immunohistochemistry (cut off point, 10%). The two methods were compared by means of kappa statistics, and the prognostic significance of both in relation to disease free survival (DFS) and overall survival (OS) was determined. RESULTS: SPF and Ki-67 analysis was performed on 234 (76.5%) and 295 (96.4%) tumours, respectively. The two assessments were simultaneously available in 230 cases. All SPF variables analysed in the whole series significantly correlated with disease evolution, with the conventional median SPF (cut off point, 6.1%) showing the highest predictive value in relation to both DFS (p = 0.0001) and OS (p = 0.0003). SPF tertiles and median SPF evaluated according to DNA ploidy status had no prognostic significance. The Ki-67 index showed a trend in relation to DFS (p = 0.086) that did not reach significance, and no correlation with OS was found (p = 0.264). The comparative analysis of SPF and Ki-67 revealed some agreement between the two methods (agreement, 69.13%; kappa statistic, 0.3844; p < 0.001), especially in the subgroup of diploid tumours. CONCLUSIONS: Flow cytometric SPF is a better prognosticator than the Ki-67 index, but only SPF variables applied in the whole series show potential clinical usefulness. PMID- 11429428 TI - Sentinel node investigation in breast cancer: detailed analysis of the yield from step sectioning and immunohistochemistry. AB - AIMS: To evaluate in detail the extent to which step sectioning and immunohistochemical examination of sentinel lymph nodes (SNs) in patients with breast cancer reveal additional node positive patients, to arrive at a sensitive yet workable protocol for histopathological SN examination. METHODS: This study comprised 86 women with one or more positive SN after a successful SN procedure for clinical stage T1-T2 invasive breast cancer. SNs were lamellated into pieces of approximately 0.5 cm in size. One initial haematoxylin and eosin (H&E) stained central cross section was made for each block. When negative, four step ribbons were cut at intervals of 250 microm. One section from each ribbon was stained with H&E, and one was used for immunohistochemistry (IHC). RESULTS: When taking the cumulative total of detected metastases at level 5 as 100%, the percentage of SN positive patients increased from 80%, 83%, 85%, 87% to 88% in the H&E sections through levels 1 to 5, and with IHC these values were 86%, 90%, 94%, 98%, and 100%. Three of nine patients in whom metastases were detected at levels 3-5 only had metastases in the subsequent axillary lymph node dissection. CONCLUSIONS: Multiple level sectioning of SNs (five levels at 250 microm intervals) and the use of IHC detects additional metastases up to the last level. Although more levels of sectioning might increase the yield even further, this protocol ensures a reasonable workload for the pathologist with an acceptable sensitivity when compared with the published literature. PMID- 11429429 TI - Salicylic acid in the serum of subjects not taking aspirin. Comparison of salicylic acid concentrations in the serum of vegetarians, non-vegetarians, and patients taking low dose aspirin. AB - AIMS: To determine serum salicylic acid concentrations in non-vegetarians and vegetarians not taking salicylate drugs, and to compare these concentrations with those found in patients taking aspirin, 75 mg daily. METHODS: Serum samples were obtained from vegetarians (n = 37) and non-vegetarians (n = 39) not taking salicylate drugs. Non-vegetarians and vegetarians were recruited from the community and from a Buddhist monastery, respectively, in Dumfries and Galloway, Scotland. Patients (n = 14) taking aspirin (75 mg daily) were recruited from the Dumfries diabetic clinic. Serum salicylic acid concentrations were determined using a high performance liquid chromatography method with electrochemical detection. RESULTS: Salicylic acid was detected in every serum sample analysed. Higher serum concentrations of salicylic acid were found in vegetarians than non vegetarians: median concentrations of 0.11 (range, 0.04-2.47) micromol/litre and 0.07 (range, 0.02-0.20) micromol/litre, respectively; the median of the difference was 0.05 micromol/litre (95% confidence interval for difference, 0.03 to 0.08; p < 0.0001). The median serum concentration of salicylic acid in patients taking aspirin (75 mg daily) was 10.03 (range, 0.23-25.40) micromol/litre, which was significantly higher than that found in non-vegetarians and vegetarians. There was overlap in serum salicylic acid concentrations between the vegetarians and patients taking aspirin. CONCLUSIONS: Salicylic acid, a non steroidal anti-inflammatory drug, is present in fruits and vegetables and is found in higher concentrations in vegetarians than non-vegetarians. This suggests that a diet rich in fruits and vegetables contributes to the presence of salicylic acid in vivo. There is overlap between the serum concentrations of salicylic acid in vegetarians and patients taking aspirin, 75 mg daily. These findings may explain, in part, the health promoting effects of dietary fruits and vegetables. PMID- 11429430 TI - Introduction of an automated service for the laboratory confirmation of meningococcal disease in Scotland. AB - The Scottish Meningococcus and Pneumococcus Reference Laboratory provides a national service for the laboratory confirmation of meningococcal and pneumococcal disease in Scotland. The main tests used for the laboratory confirmation of meningococcal disease are culture, the polymerase chain reaction (PCR), antibody testing, and more recently DNA sequencing. This paper describes the automation of PCR for the laboratory confirmation of meningococcal disease and the typing of meningococcal isolates using DNA sequencing. Both methods have been automated using a robotic liquid handler and automated DNA sequencer. These methods, along with standard culture phenotyping and antibody testing, provide Scotland with an excellent service for the confirmation of meningococcal disease. PMID- 11429431 TI - No sequence variation in part of the hexon and the fibre genes of adenovirus 8 isolated from patients with conjunctivitis or epidemic keratoconjunctivitis (EKC) in Norway during 1989 to 1996. AB - BACKGROUND/AIMS: Several local epidemics of keratoconjunctivitis/conjunctivitis caused by adenovirus type 8 (Ad8) occurred in Norway from August 1995 to May 1996. A smaller epidemic occurred in 1992. The Ad8 hexon forms the surface of the virion and contains the hypervariable regions loop I(1) and loop I(2). The fibre mediates the primary contact with cells. Sequence variation in hexon and fibre genes might play an important role in the pathogenicity of adenoviruses. The aim of this study was to investigate the genetic variability at the hexon and fibre genes in 26 strains of Ad8 isolated from 1989 to 1996. METHODS: The genetic variability of 26 strains of Ad8 isolated from 1989 to 1996 was studied by sequencing part of the hexon and fibre genes. The Ad8 sequences were compared with each other and with two Ad8 strains from the EMBL database. In addition, 14 of the 26 isolates were subjected to restriction endonuclease analysis. RESULTS: No significant sequence variation was seen during the six year period. CONCLUSION: The Ad8 strains causing epidemics of keratoconjunctivitis/conjunctivitis in Norway are genetically stable. PMID- 11429432 TI - Lymphadenopathy and lymph node infarction as a result of gold injections. AB - This report describes a case of lymphadenopathy and lymph node infarction as a consequence of intramuscular gold administered to a patient suffering from rheumatoid arthritis, to highlight this rare association. A 34 year old woman with a four year history of rheumatoid arthritis affecting multiple joints was started on intramuscular gold injections after little response to anti inflammatory medication. After her sixth injection the patient developed enlarged neck and axillary lymph nodes. Biopsy showed subtotal infarction of a reactive node, confirmed by histochemical, immunohistochemical, and molecular techniques. The patient continued to suffer from rheumatoid arthritis with no evidence of malignant lymphoma after three years. This case provides strong evidence that lymphadenopathy with infarction is a rare complication of gold injections. In such a situation, it is particularly important to exclude a diagnosis of lymphoma, because this is the most common cause of spontaneous lymph node infarction. This can be achieved through awareness of the association, and by the use of ancillary histochemical, immunohistochemical, and molecular techniques on the biopsy material. PMID- 11429433 TI - The role of molecular analysis of immunoglobulin and T cell receptor gene rearrangements in the diagnosis of lymphoproliferative disorders. AB - AIMS: To investigate whether the analysis of immunoglobulin (Ig)/T cell receptor (TCR) rearrangements is useful in the diagnosis of lymphoproliferative disorders. METHODS: In a series of 107 consecutive cases with initial suspicion of non Hodgkin's lymphoma (NHL), Southern blot (SB) analysis of Ig/TCR rearrangements was performed. RESULTS: In 98 of 100 histopathologically conclusive cases, Ig/TCR gene results were concordant. In one presumed diffuse large B cell lymphoma (DLCL) and one follicular lymphoma (FL) case no clonality could be detected by SB analysis, or by polymerase chain reaction (PCR) at second stage. In the DLCL, sampling error might have occurred; the FL was revised after an initial diagnosis of reactivity. In many of the histopathologically inconclusive cases Ig/TCR gene SB analysis was helpful, giving support for the histopathological suspicion. However, because of a lack of (clinical) follow up data this could not be confirmed in a few cases. CONCLUSIONS: Experienced haematopathologists or a pathologist panel can diagnose malignant versus reactive lesions in most cases without the need for Ig/TCR gene analysis and can select the 5-10% of cases that might benefit from molecular clonality studies. PMID- 11429434 TI - Predominant fatty variant of myofibroblastoma of breast. AB - Myofibroblastoma of the breast is an uncommon but well defined benign stromal tumour. This report describes a case in which the predominant histological component was mature adipose tissue and two further cases with a major adipocytic component. Although small foci of adipose tissue are a recognised feature of this tumour, the dominance of the histological pattern by fat has not been described previously, and the recognition of this variant is important to allow confident diagnosis and avoid confusion with other primary adipocytic or stromal lesions, especially in the setting of potential needle core biopsy of such a lesion. PMID- 11429435 TI - Sarcoma of the thyroid region mimicking Riedel's thyroiditis. AB - Because sarcomas of the anterior lower neck region occur so infrequently, they are not usually considered in the differential diagnosis of Riedel's thyroiditis. Riedel's thyroiditis itself may be confused on clinical grounds alone with malignant neoplasms because of its invasive features. Sarcomatoid carcinoma is the main entity to be discarded in this regard. This is accomplished through histological examination by the finding of carcinomatous areas and/or reactivity with epithelial markers. These features also set apart sarcomatoid carcinoma from true sarcomas. This report concerns a patient with a sarcoma of the anterior lower neck region which was initially suspected to be Riedel's thyroiditis or sarcomatoid carcinoma on clinical and radiological grounds. A peroperative biopsy was interpreted by two independent pathologists as consistent with Riedel's thyroiditis. The subsequent clinical course and postmortem examination demonstrated a high grade sarcoma with metastasis to both lungs and the pleura, and invasion of adjacent neck structures. Nevertheless, some areas of the postmortem material showed a microscopic pattern similar to mediastinal fibrosis, raising the possibility of the malignant transformation of a fibrosclerotic lesion. PMID- 11429436 TI - Down's syndrome screening: a controversial test, with more controversy to come! PMID- 11429437 TI - Microsatellite unstable colorectal cancer. PMID- 11429438 TI - Scientific dishonesty: European reflections. PMID- 11429439 TI - S4 charges move close to residues in the pore domain during activation in a K channel. AB - Voltage-gated ion channels respond to changes in the transmembrane voltage by opening or closing their ion conducting pore. The positively charged fourth transmembrane segment (S4) has been identified as the main voltage sensor, but the mechanisms of coupling between the voltage sensor and the gates are still unknown. Obtaining information about the location and the exact motion of S4 is an important step toward an understanding of these coupling mechanisms. In previous studies we have shown that the extracellular end of S4 is located close to segment 5 (S5). The purpose of the present study is to estimate the location of S4 charges in both resting and activated states. We measured the modification rates by differently charged methanethiosulfonate regents of two residues in the extracellular end of S5 in the Shaker K channel (418C and 419C). When S4 moves to its activated state, the modification rate by the negatively charged sodium (2 sulfonatoethyl) methanethiosulfonate (MTSES(-)) increases significantly more than the modification rate by the positively charged [2-(trimethylammonium)ethyl] methanethiosulfonate, bromide (MTSET(+)). This indicates that the positive S4 charges are moving close to 418C and 419C in S5 during activation. Neutralization of the most external charge of S4 (R362), shows that R362 in its activated state electrostatically affects the environment at 418C by 19 mV. In contrast, R362 in its resting state has no effect on 418C. This suggests that, during activation of the channel, R362 moves from a position far away (>20 A) to a position close (8 A) to 418C. Despite its close approach to E418, a residue shown to be important in slow inactivation, R362 has no effect on slow inactivation or the recovery from slow inactivation. This refutes previous models for slow inactivation with an electrostatic S4-to-gate coupling. Instead, we propose a model with an allosteric mechanism for the S4-to-gate coupling. PMID- 11429441 TI - Electrically triggered all-or-none Ca(2)+-liberation during action potential in the giant alga Chara. AB - Electrically triggered action potentials in the giant alga Chara corallina are associated with a transient rise in the concentration of free Ca(2)+ in the cytoplasm (Ca(2)+(cyt)). The present measurements of Ca(2)+(cyt) during membrane excitation show that stimulating pulses of low magnitude (subthreshold pulse) had no perceivable effect on Ca(2)+(cyt). When the strength of a pulse exceeded a narrow threshold (suprathreshold pulse) it evoked the full extent of the Ca(2)+(cyt) elevation. This suggests an all-or-none mechanism for Ca(2)+ mobilization. A transient calcium rise could also be induced by one subthreshold pulse if it was after another subthreshold pulse of the same kind after a suitable interval, i.e., not closer than a few 100 ms and not longer than a few seconds. This dependency of Ca(2)+ mobilization on single and double pulses can be simulated by a model in which a second messenger is produced in a voltage dependent manner. This second messenger liberates Ca(2)+ from internal stores in an all-or-none manner once a critical concentration (threshold) of the second messenger is exceeded in the cytoplasm. The positive effect of a single suprathreshold pulse and two optimally spaced subthreshold pulses on Ca(2)+ mobilization can be explained on the basis of relative velocity for second messenger production and decomposition as well as the availability of the precursor for the second messenger production. Assuming that inositol-1,4,5, trisphosphate (IP(3)) is the second messenger in question, the present data provide the major rate constants for IP(3) metabolism. PMID- 11429442 TI - Different fast-gate regulation by external Cl(-) and H(+) of the muscle-type ClC chloride channels. AB - The fast gate of the muscle-type ClC channels (ClC-0 and ClC-1) opens in response to the change of membrane potential (V). This gating process is intimately associated with the binding of external Cl(-) to the channel pore in a way that the occupancy of Cl(-) on the binding site increases the channel's open probability (P(o)). External H(+) also enhances the fast-gate opening in these channels, prompting a hypothesis that protonation of the binding site may increase the Cl(-) binding affinity, and this is possibly the underlying mechanism for the H(+) modulation. However, Cl(-) and H(+), modulate the fast gate P(o)-V curve in different ways. Varying the external Cl(-) concentrations ([Cl(-)](o)) shifts the P(o)-V curve in parallel along the voltage axis, whereas reducing external pH mainly increases the minimal P(o) of the curve. Furthermore, H(+) modulations at saturating and nonsaturating [Cl(-)](o) are similar. Thus, the H(+) effect on the fast gating appears not to be a consequence of an increase in the Cl(-) binding affinity. We previously found that a hyperpolarization favored opening process is important to determine the fast-gate P(o) of ClC-0 at very negative voltages. This [Cl(-)](o)-independent mechanism attracted little attention, but it appears to be the opening process that is modulated by external H(+). PMID- 11429443 TI - Molecular basis of Ca(2)+ activation of the mouse cardiac Ca(2)+ release channel (ryanodine receptor). AB - Activation of the cardiac ryanodine receptor (RyR2) by Ca(2)+ is an essential step in excitation-contraction coupling in heart muscle. However, little is known about the molecular basis of activation of RyR2 by Ca(2)+. In this study, we investigated the role in Ca(2)+ sensing of the conserved glutamate 3987 located in the predicted transmembrane segment M2 of the mouse RyR2. Single point mutation of this conserved glutamate to alanine (E3987A) reduced markedly the sensitivity of the channel to activation by Ca(2)+, as measured by using single channel recordings in planar lipid bilayers and by [(3)H]ryanodine binding assay. However, this mutation did not alter the affinity of [(3)H]ryanodine binding and the single-channel conductance. In addition, the E3987A mutant channel was activated by caffeine and ATP, was inhibited by Mg(2)+, and was modified by ryanodine in a fashion similar to that of the wild-type channel. Coexpression of the wild-type and mutant E3987A RyR2 proteins in HEK293 cells produced individual single channels with intermediate sensitivities to activating Ca(2)+. These results are consistent with the view that glutamate 3987 is a major determinant of Ca(2)+ sensitivity to activation of the mouse RyR2 channel, and that Ca(2)+ sensing by RyR2 involves the cooperative action between ryanodine receptor monomers. The results of this study also provide initial insights into the structural and functional properties of the mouse RyR2, which should be useful for studying RyR2 function and regulation in genetically modified mouse models. PMID- 11429444 TI - In vivo assessment of local phosphodiesterase activity using tailored cyclic nucleotide-gated channels as cAMP sensors. AB - Phosphodiesterases (PDEs) catalyze the hydrolysis of the second messengers cAMP and cGMP. However, little is known about how PDE activity regulates cyclic nucleotide signals in vivo because, outside of specialized cells, there are few methods with the appropriate spatial and temporal resolution to measure cyclic nucleotide concentrations. We have previously demonstrated that adenovirus expressed, olfactory cyclic nucleotide-gated channels provide real-time sensors for cAMP produced in subcellular compartments of restricted diffusion near the plasma membrane (Rich, T.C., K.A. Fagan, H. Nakata, J. Schaack, D.M.F. Cooper, and J.W. Karpen. 2000. J. Gen. Physiol. 116:147-161). To increase the utility of this method, we have modified the channel, increasing both its cAMP sensitivity and specificity, as well as removing regulation by Ca(2)+-calmodulin. We verified the increased sensitivity of these constructs in excised membrane patches, and in vivo by monitoring cAMP-induced Ca(2)+ influx through the channels in cell populations. The improved cAMP sensors were used to monitor changes in local cAMP concentration induced by adenylyl cyclase activators in the presence and absence of PDE inhibitors. This approach allowed us to identify localized PDE types in both nonexcitable HEK-293 and excitable GH4C1 cells. We have also developed a quantitative framework for estimating the K(I) of PDE inhibitors in vivo. The results indicate that PDE type IV regulates local cAMP levels in HEK-293 cells. In GH4C1 cells, inhibitors specific to PDE types I and IV increased local cAMP levels. The results suggest that in these cells PDE type IV has a high K(m) for cAMP, whereas PDE type I has a low K(m) for cAMP. Furthermore, in GH4C1 cells, basal adenylyl cyclase activity was readily observable after application of PDE type I inhibitors, indicating that there is a constant synthesis and hydrolysis of cAMP in subcellular compartments near the plasma membrane. Modulation of constitutively active adenylyl cyclase and PDE would allow for rapid control of cAMP-regulated processes such as cellular excitability. PMID- 11429445 TI - Contrasting contributions of endoplasmic reticulum and mitochondria to Ca(2)+ handling in neurons. PMID- 11429446 TI - Multiple modes of calcium-induced calcium release in sympathetic neurons I: attenuation of endoplasmic reticulum Ca2+ accumulation at low [Ca2+](i) during weak depolarization. AB - Many cells express ryanodine receptors (RyRs) whose activation is thought to amplify depolarization-evoked elevations in cytoplasmic Ca2+ concentration [Ca2+](i) through a process of Ca2+ -induced Ca2+ release (CICR). In neurons, it is usually assumed that CICR triggers net Ca2+ release from an ER Ca2+ store. However, since net ER Ca 2+ transport depends on the relative rates of Ca2+ uptake and release via distinct pathways, weak activation of a CICR pathway during periods of ER Ca accumulation would have a totally different effect: attenuation of Ca2+ accumulation. Stronger CICR activation at higher [Ca2+](i) could further attenuate Ca2+ accumulation or trigger net Ca2+ release, depending on the quantitative properties of the underlying Ca2+ transporters. This and the companion study (Hongpaisan, J., N.B. Pivovarova, S.L. Colgrove, R.D. Leapman, and D.D. Friel, and S.B. Andrews. 2001. J. Gen. Physiol. 118:101-112) investigate which of these CICR "modes" operate during depolarization-induced Ca2+ entry in sympathetic neurons. The present study focuses on small [Ca2+](i) elevations (less than approximately 350 nM) evoked by weak depolarization. The following two approaches were used: (1) Ca2+ fluxes were estimated from simultaneous measurements of [Ca2+](i) and I(Ca) in fura-2-loaded cells (perforated patch conditions), and (2) total ER Ca concentrations ([Ca](ER)) were measured using X ray microanalysis. Flux analysis revealed triggered net Ca2+ release during depolarization in the presence but not the absence of caffeine, and [Ca2+](i) responses were accelerated by SERCA inhibitors, implicating ER Ca2+ accumulation, which was confirmed by direct [Ca](ER) measurements. Ryanodine abolished caffeine induced CICR and enhanced depolarization-induced ER Ca2+ accumulation, indicating that activation of the CICR pathway normally attenuates ER Ca2+ accumulation, which is a novel mechanism for accelerating evoked [Ca2+](i) responses. Theory shows how such a low gain mode of CICR can operate during weak stimulation and switch to net Ca2+ release at high [Ca2+](i), a transition demonstrated in the companion study. These results emphasize the importance of the relative rates of Ca2+ uptake and release in defining ER contributions to depolarization-induced Ca2+ signals. PMID- 11429447 TI - Multiple modes of calcium-induced calcium release in sympathetic neurons II: a [Ca2+](i)- and location-dependent transition from endoplasmic reticulum Ca accumulation to net Ca release. AB - CICR from an intracellular store, here directly characterized as the ER, usually refers to net Ca(2)+ release that amplifies evoked elevations in cytosolic free calcium [Ca2+](i). However, the companion paper (Albrecht, M.A., S.L. Colegrove, J. Hongpaisan, N.B. Pivovarova, S.B. Andrews, and D.D. Friel. 2001. J. Gen. Physiol. 118:83-100) shows that in sympathetic neurons, small [Ca2+](i) elevations evoked by weak depolarization stimulate ER Ca accumulation, but at a rate attenuated by activation of a ryanodine-sensitive CICR pathway. Here, we have measured depolarization-evoked changes in total ER Ca concentration ([Ca](ER)) as a function of [Ca2+](i), and found that progressively larger [Ca2+](i) elevations cause a graded transition from ER Ca accumulation to net release, consistent with the expression of multiple modes of CICR. [Ca](ER) is relatively high at rest (12.8 +/- 0.9 mmol/kg dry weight, mean +/- SEM) and is reduced by thapsigargin or ryanodine (5.5 +/- 0.7 and 4.7 +/- 1.1 mmol/kg, respectively). [Ca](ER) rises during weak depolarization (to 17.0 +/- 1.6 mmol/kg over 120s, [Ca2+](i) less than approximately 350 nM), changes little in response to stronger depolarization (12.1 +/- 1.1 mmol/kg, [Ca2+](i) approximately 700 nM), and declines (to 6.5 +/- 1.0 mmol/kg) with larger [Ca2+](i) elevations (>1 microM) evoked by the same depolarization when mitochondrial Ca2+ uptake is inhibited (FCCP). Thus, net ER Ca2+ transport exhibits a biphasic dependence on [Ca2+](i). With mitochondrial Ca2+ uptake enabled, [Ca](ER) rises after repolarization (to 16.6 +/- 1.8 mmol/kg at 15 min) as [Ca2+](i) falls within the permissive range for ER Ca accumulation over a period lengthened by mitochondrial Ca2+ release. Finally, although spatially averaged [Ca](ER) is unchanged during strong depolarization, net ER Ca2+ release still occurs, but only in the outermost approximately 5-microm cytoplasmic shell where [Ca2+](i) should reach its highest levels. Since mitochondrial Ca accumulation occurs preferentially in peripheral cytoplasm, as demonstrated here by electron energy loss Ca maps, the Ca content of ER and mitochondria exhibit reciprocal dependencies on proximity to sites of Ca2+ entry, possibly reflecting indirect mitochondrial regulation of ER Ca(2)+ transport. PMID- 11429448 TI - Selective open-channel block of Shaker (Kv1) potassium channels by s nitrosodithiothreitol (SNDTT). AB - Large quaternary ammonium (QA) ions block voltage-gated K(+) (Kv) channels by binding with a 1:1 stoichiometry in an aqueous cavity that is exposed to the cytoplasm only when channels are open. S-nitrosodithiothreitol (SNDTT; ONSCH(2)CH(OH)CH(OH)CH(2)SNO) produces qualitatively similar "open-channel block" in Kv channels despite a radically different structure. SNDTT is small, electrically neutral, and not very hydrophobic. In whole-cell voltage-clamped squid giant fiber lobe neurons, bath-applied SNDTT causes reversible time dependent block of Kv channels, but not Na(+) or Ca(2)+ channels. Inactivation removed ShakerB (ShBDelta) Kv1 channels expressed in HEK 293 cells are similarly blocked and were used to study further the action of SNDTT. Dose-response data are consistent with a scheme in which two SNDTT molecules bind sequentially to a single channel, with binding of the first being sufficient to produce block. The dissociation constant for the binding of the second SNDTT molecule (K(d2) = 0.14 mM) is lower than that of the first molecule (K(d1) = 0.67 mM), indicating cooperativity. The half-blocking concentration (K(1/2)) is approximately 0.2 mM. Steady-state block by this electrically neutral compound has a voltage dependence (about -0.3 e(0)) similar in magnitude but opposite in directionality to that reported for QA ions. Both nitrosyl groups on SNDTT (one on each sulfur atom) are required for block, but transfer of these reactive groups to channel cysteine residues is not involved. SNDTT undergoes a slow intramolecular reaction (tau approximately 770 s) in which these NO groups are liberated, leading to spontaneous reversal of the SNDTT effect. Competition with internal tetraethylammonium indicates that bath-applied SNDTT crosses the cell membrane to act at an internal site, most likely within the channel cavity. Finally, SNDTT is remarkably selective for Kv1 channels. When individually expressed in HEK 293 cells, rat Kv1.1-1.6 display profound time-dependent block by SNDTT, an effect not seen for Kv2.1, 3.1b, or 4.2. PMID- 11429449 TI - Out of the iron age: new insights into the critical role of manganese homeostasis in bacteria. PMID- 11429450 TI - Repeated ruminal dosing of Ruminococcus spp. does not result in persistence, but changes in other microbial populations occur that can be measured with quantitative 16S-rRNA-based probes. AB - Digestibility of fibre in ruminants may be improved by the introduction of highly fibrolytic strains of ruminal bacteria. This approach may be feasible if, for example, strains of Ruminococcus that are significantly more fibrolytic than the normal population of Ruminococcus are used for inoculation purposes. Introduced strains of bacteria, irrespective of ecosystem, often decline after inoculation, and in this study, highly fibrolytic strains of Ruminococcus were continuously dosed to ensure that measurements of fibre digestion were made in the presence of significant numbers of the introduced bacteria. During dosing the total culturable count increased significantly (P<0.05), but declined post-dosing. The level of dosed Ruminococcus, and total Ruminococcus, Fibrobacter succinogenes and eukaryotes measured by 16S rRNA probes increased significantly (P<0.05) during the dosing period, but also declined post-dosing. When in vitro nylon bag digestibility, feed intake or whole-tract digestibility was measured, no improvement could be measured. PMID- 11429451 TI - Closely related Prochlorococcus genotypes show remarkably different depth distributions in two oceanic regions as revealed by in situ hybridization using 16S rRNA-targeted oligonucleotides. AB - An in situ hybridization method was applied to the identification of marine cyanobacteria assignable to the genus Prochlorococcus using horseradish peroxidase-labelled 16S rRNA-targeted oligonucleotide probes in combination with tyramide signal amplification (TSA). With this method very bright signals were obtained, in contrast to hybridizations with oligonucleotides monolabelled with fluorochromes, which failed to give positive signals. Genotype-specific oligonucleotides for high light (HL)- and low light (LL)-adapted members of this genus were identified by 16S rRNA sequence analyses and their specificities confirmed in whole-cell hybridizations with cultured strains of Prochlorococcus marinus Chisholm et al., 1992, Prochlorococcus sp. and Synechococcus sp. In situ hybridization of these genotype-specific probes to field samples from stratified water bodies collected in the North Atlantic Ocean and the Red Sea allowed a rapid assessment of the abundance and spatial distribution of HL- and LL-adapted Prochlorococcus. In both oceanic regions the LL-adapted Prochlorococcus populations were localized in deeper water whereas the HL-adapted Prochlorococcus populations were not only distinct in each region but also exhibited strikingly different depth distributions, HLI being confined to shallow water in the North Atlantic, in contrast to HLII, which was present throughout the water column in the Red Sea. PMID- 11429452 TI - Use of a halobacterial bgaH reporter gene to analyse the regulation of gene expression in halophilic archaea. AB - The bgaH reading frame encoding a beta-galactosidase of 'Haloferax alicantei' was used as a reporter gene to investigate three different promoter regions derived from gvpA genes of Haloferax mediterranei (mc-gvpA) and Halobacterium salinarum (c-gvpA and p-gvpA) in Haloferax volcanii transformants. The fusion of bgaH at the start codon of each gvpA reading frame (A1-bgaH fusion genes) caused translational problems in some cases. Transformants containing constructs with fusions further downstream in the gvpA reading frame (A-bgaH) produced beta galactosidase, and colonies on agar plates turned blue when sprayed with X-Gal. The beta-galactosidase activities quantified by standard ONPG assays correlated well with the mRNA data determined with transformants containing the respective gvpA genes: the cA-bgaH fusion gene was completely inactive, the mcA-bgaH transformants showed low amounts of products, whereas the pA-bgaH fusion gene was constitutively expressed in the respective transformants. The transcription of each A-bgaH gene was activated by the homologous transcriptional activator protein GvpE. The cGvpE, pGvpE and mcGvpE proteins were able to activate the promoter of pA-bgaH and mcA-bgaH, whereas the promoter of cA-bgaH was only activated by cGvpE. Among the three GvpE proteins tested, cGvpE appeared to be the strongest transcriptional activator. PMID- 11429453 TI - Differential regulation of laccase gene expression in Pleurotus sajor-caju. AB - Four laccase isozyme genes, Psc lac1, 2, 3 and 4 have been cloned from the edible mushroom, Pleurotus sajor-caju. The genes display a high degree of homology with other basidiomycete laccases (55-99%) at the amino acid level. Of the laccase genes isolated, Psc lac1 and 4 displayed the highest degree of similarity (85% at the amino acid level), while Psc lac3 showed the highest degree of divergence, exhibiting only 52-57% amino acid similarity to the other PL: sajor-caju laccase gene sequences. Laccase activity in PL: sajor-caju is affected by nutrient nitrogen and carbon, and by the addition of copper and manganese to the growth medium. In addition, 2,5-xylidine, ferulic acid, veratric acid and 1 hydroxybenzotriazole induced laccase activity in the fungus. Induction of individual laccase isozyme genes by carbon, nitrogen, copper, manganese and the two aromatic compounds, 2,5-xylidine and ferulic acid, occurred at the level of gene transcription. While Psc lac3 transcript levels appeared to be constitutively expressed, transcript levels for the other laccase isozyme genes, lac1, 2 and 4, were differentially regulated under the conditions tested. PMID- 11429454 TI - Expression control and specificity of the basic amino acid exporter LysE of Corynebacterium glutamicum. AB - LysE of Corynebacterium glutamicum belongs to a large new superfamily of translocators whose members are probably all involved in the export of small solutes. Here, the transcript initiation site of lysE, and its divergently transcribed regulator gene, lysG, are identified. Single-copy transcriptional fusions of lysE with lacZ, and titration experiments, show that LysG is the positive regulator of lysE expression enabling its up to 20-fold induction. This induction requires the presence of a coinducer, which is either intracellular L lysine, or L-arginine. A competition experiment showed that LysE exports these two basic amino acids at comparable rates of about 0.75 nmol min(-1) (mg dry wt)( 1). Although L-histidine and L-citrulline also act as coinducers of lysE expression, these two amino acids are not exported by LysE. As is evident from the analysis of a lysEG deletion mutant, the physiological role of the lysEG system is to prevent bacteriostasis due to elevated L-lysine or L-arginine concentrations that arise during growth in the presence of peptides or in mutants possessing a deregulated biosynthesis pathway. C. glutamicum has additional export activities other than those of LysE for exporting L-histidine, L citrulline and L-ornithine. PMID- 11429455 TI - Bacterial Ohr and OsmC paralogues define two protein families with distinct functions and patterns of expression. AB - Xanthomonas campestris Ohr (a protein involved in organic peroxide protection) and Escherichia coli OsmC (an osmotically inducible protein of unknown function) are related proteins. Database searches and phylogenetic analyses reveal that Ohr and OsmC homologues cluster into two related subfamilies of proteins widely distributed in both Gram-negative and Gram-positive bacteria. To determine if these two subfamilies are functionally distinct, ohr and osmC in Pseudomonas aeruginosa (a bacterium with one representative from each subfamily) were analysed. Only ohr mutants are hypersensitive to organic peroxide, and this phenotype can be restored by complementation with ohr but not osmC. In addition, expression of ohr was highly induced only by organic peroxides, and not by other oxidants or stresses. In contrast, osmC was induced by ethanol and osmotic stress. A similar pattern of regulation was observed for Ohr and OsmC homologues in the Gram-positive bacterium Deinococcus radiodurans, though uninduced expression was much higher and induction lower in this species. These data clearly support the conclusion that Ohr and OsmC define two functionally distinct subfamilies with distinct patterns of regulation. PMID- 11429456 TI - YkrB is the main peptide deformylase in Bacillus subtilis, a eubacterium containing two functional peptide deformylases. AB - Peptide deformylation is an essential process in eubacteria. The peptide deformylase Def has been suggested to be an attractive target for antibacterial drug discovery. Some eubacteria including medically important pathogens possess two def-like genes. Until now, the functionality of both genes has been tested only in Staphylococcus aureus with the result that one gene copy was functional. Here, expression of two functional def-like gene products in Bacillus subtilis is demonstrated. Besides the def gene, which is chromosomally located close to the formyltransferase gene fmt and which was overexpressed and biochemically tested previously, B. subtilis possesses a second def-like gene, called ykrB. The encoded protein is 32% identical to the def gene product. It was shown that either def or ykrB had to be present for growth of B. subtilis in rich medium (each was individually dispensable). Studies with a def/ykrB double deletion strain with xylose-inducible ykrB copy demonstrated that, besides def, the gene ykrB is a second cellular target of deformylase inhibitors such as the antibiotic actinonin. The gene products exhibited similar enzymic properties, exemplified by similar inhibition efficacy of actinonin in biochemical assays. Antibiotic susceptibility tests with different deletion strains and Northern analyses indicated that YkrB is probably the predominant deformylase in B. subtilis. It was shown that duplication of the deformylase function does not lead to an increased actinonin-resistance frequency in comparison to B. subtilis mutants carrying only one deformylase gene. PMID- 11429457 TI - A Bacillus amyloliquefaciens ChbB protein binds beta- and alpha-chitin and has homologues in related strains. AB - A small (19.8 kDa) protein was identified in Bacillus amyloliquefaciens ALKO 2718 cultures during growth in the presence of yeast extract and chitin, but not with glucose. The protein targets beta-chitin best, then alpha-chitin, but barely any other polysaccharide. This described chitin-binding protein (ChbB) is the first of its type from a Bacillus strain and cross-reacts with antibodies raised against the Streptomyces alpha-chitin-binding protein CHB1. Using reverse genetics, the chromosomal chbB gene of strain ALKO 2718 was identified, cloned and sequenced. ChbB shares several motifs with the alpha-chitin-binding proteins CHB1 and CHB2 of Streptomyces and CBP21 of Serratia marcescens predominantly targeting beta-chitin. Synthesis was repressed by glucose and the presence of cre boxes suggests catabolite control. Using PCR, Southern hybridization and anti ChbB antibodies, the presence of a chbB gene, as well as of a ChbB protein homologue, was ascertained in several tested B. amyloliquefaciens strains, but not in Bacillus subtilis 168. Contrary to B. subtilis 168, all B. amyloliquefaciens strains secreted varying amounts of enzymic activity, degrading carboxymethyl chitin coupled with Remazol brilliant violet. PMID- 11429458 TI - Identification of genes involved in the activation of the Bacillus thuringiensis inhA metalloprotease gene at the onset of sporulation. AB - The immune inhibitor A (InhA) metalloprotease from Bacillus thuringiensis specifically cleaves antibacterial proteins produced by the insect host, suggesting that it may contribute to the overall virulence of B. thuringiensis. The transcriptional regulation of the inhA gene in both B. thuringiensis and Bacillus subtilis was investigated. Using a transcriptional inhA'-lacZ fusion, it was shown that inhA expression is activated at the onset of sporulation. However, the transcriptional start site of inhA is similar to sigma(A)-dependent promoters, and deletion of the sporulation-specific sigma factors sigma(F) or sigma(E) had no effect on inhA expression in B. subtilis. The DNA region upstream from inhA contains two genes encoding polypeptides similar to the SinI and SinR regulators of B. subtilis. SinR is a DNA-binding protein regulating gene expression and SinI inhibits SinR activity. Overexpression of the sin genes affects the expression of the inhA'-lacZ transcriptional fusion in B. thuringiensis: early induction of inhA expression was observed when sinI was overexpressed, whereas inhA expression was reduced in a strain overexpressing sinR, suggesting that inhA transcription is repressed, directly or indirectly, by SinR. inhA transcription was greatly reduced in B. thuringiensis and B. subtilis spo0A mutants. Analysis of the inhA'-lacZ expression in abrB and abrB-spo0A mutants of B. subtilis indicates that the Spo0A-dependent regulation of inhA expression depends on AbrB, which is known to regulate expression of transition state and sporulation genes in B. subtilis. PMID- 11429459 TI - Genetic and biochemical characterization of an enantioselective amidase from Agrobacterium tumefaciens strain d3. AB - An enantioselective amidase was purified to homogeneity from Agrobacterium tumefaciens d3. The enzyme has a molecular mass of about 490000 Da and is composed of identical subunits with a molecular mass of about 63000 Da. The purified enzyme converted racemic 2-phenylpropionamide to the corresponding S acid with an enantiomeric excess (ee) value >95% at almost 50% conversion of the racemic amide. The purified enzyme was digested with trypsin and the amino acid sequences of the N terminus and different tryptic peptides determined. These amino acid sequences were used to clone the encoding gene. Finally, a 9330 bp DNA fragment was sequenced and the amidase gene identified. The deduced amino acid sequence showed homology to other enantioselective amidases from different bacterial genera. No indications of a structural coupling of the amidase gene with the genes for a nitrile hydratase could be found on the cloned DNA fragment. The amidase gene was encoded by an approximately 500 kb circular plasmid in A. tumefaciens d3. The amidase was heterologously expressed in Escherichia coli and, as well as 2-phenylpropionamide, was shown to hydrolyse alpha-chloro- and alpha methoxyphenylacetamide and 2-methyl-3-phenylpropionamide highly enantioselectively. Some amino acids within a highly conserved region common amongst all known enantioselective amidases ('amidase signature') were changed by site-specific mutagenesis and significant changes in the relative activities with different amides observed. PMID- 11429460 TI - Separation and characterization of individual mycolic acids in representative mycobacteria. AB - Total mycolic acid methyl ester fractions were isolated from 24 representatives of Mycobacterium tuberculosis, Mycobacterium bovis (including BCG), Mycobacterium microti, Mycobacterium kansasii and Mycobacterium avium. The total mycolate functional group composition was estimated from (1)H-NMR spectra. Mycolates were separated into alpha-mycolates, methoxymycolates and ketomycolates and each class was further separated by argentation chromatography into mycolates with no double bonds, with one trans-double bond and with one cis-double bond. Mass spectrometry revealed the mycolate chain lengths and (1)H-NMR the cis- and trans-double bond and cyclopropane ring content. The same species had similar mycolate profiles; the major type of each class had cis- or trans-cyclopropane rings and lacked double bonds. Minor proportions of possible unsaturated precursors of the cyclopropane mycolates were commonly encountered. Among unusual alpha-mycolates, many strains had tricyclopropyl components with chains extended by 6 to 8 carbons. Significantly, M. tuberculosis (Canetti) and M. avium had alpha mycolates with a trans-double bond and cyclopropane ring, whose chain lengths suggested a relationship to possible precursors of oxygenated mycolates. The methoxy- and ketomycolates from a majority of M. tuberculosis strains had minor amounts of components with additional cyclopropane rings, some of whose chains were also extended by 6 to 8 carbons. These latter mycolates were major components in the attenuated M. tuberculosis H37Ra strain, whose mycolate profile was distinct from those of other strains of M. tuberculosis. PMID- 11429461 TI - Comparison of cell-wall polysaccharides from Nectria cinnabarina with those from the group of Nectria with Sesquicillium anamorphs. AB - Alkali-extractable and water-soluble polysaccharides were purified from cell walls of five species of Sesquicillium or its teleomorphs, Nectria lasiacidis and Nectria impariphialis, and from Nectria cinnabarina, the type species of Nectria, a heterogeneous genus that belongs to the Hypocreales. Methylation and NMR analyses for determination of linkage types and structure were performed and indicated differences between the polysaccharides purified during the present study and those isolated from other nectrioid fungi, namely the presence of 5-O substituted galactofuranose (-->5)-Galf-(1-->) in the main chain together with 2,6-di-O-substituted galactofuranose (-->2,6)-Galf-(1-->) residues in Sesquicillium buxi and Sesquicillium pseudosetosum. The polysaccharide from N. impariphialis was similar to those obtained from the above species, although an additional residue of 6-O-substituted glucopyranose (-->6)-Glcp-(1-->), was detected in some side chains. In N. lasiacidis and Sesquicillium candelabrum the polysaccharide contained an additional branching point of 5,6-di-O-substituted galactofuranose (-->5,6)-Galf-(1-->) linked to terminal N-acetylglucosamine GlcNAc-(1-->). These chains were linked to a small mannan core. All these polysaccharides showed major differences to the polysaccharide of N. cinnabarina, which was formed by a main chain of (1-->6)-beta-linked galactofuranose units almost fully branched at positions 2-O by either single residues of glucopyranose or acidic chains containing glucuronic acid and mannose. PMID- 11429462 TI - Trehalose is required for the acquisition of tolerance to a variety of stresses in the filamentous fungus Aspergillus nidulans. AB - Trehalose is a non-reducing disaccharide found at high concentrations in Aspergillus nidulans conidia and rapidly degraded upon induction of conidial germination. Furthermore, trehalose is accumulated in response to a heat shock or to an oxidative shock. The authors have characterized the A. nidulans tpsA gene encoding trehalose-6-phosphate synthase, which catalyses the first step in trehalose biosynthesis. Expression of tpsA in a Saccharomyces cerevisiae tps1 mutant revealed that the tpsA gene product is a functional equivalent of the yeast Tps1 trehalose-6-phosphate synthase. The A. nidulans tpsA-null mutant does not produce trehalose during conidiation or in response to various stress conditions. While germlings of the tpsA mutant show an increased sensitivity to moderate stress conditions (growth at 45 degrees C or in the presence of 2 mM H(2)O(2)), they display a response to severe stress (60 min at 50 degrees C or in the presence of 100 mM H(2)O(2)) similar to that of wild-type germlings. Furthermore, conidia of the tpsA mutant show a rapid loss of viability upon storage. These results are consistent with a role of trehalose in the acquisition of stress tolerance. Inactivation of the tpsA gene also results in increased steady-state levels of sugar phosphates but does not prevent growth on rapidly metabolizable carbon sources (glucose, fructose) as seen in Saccharomyces cerevisiae. This suggests that trehalose 6-phosphate is a physiological inhibitor of hexokinase but that this control is not essential for proper glycolytic flux in A. nidulans. Interestingly, tpsA transcription is not induced in response to heat shock or during conidiation, indicating that trehalose accumulation is probably due to a post-translational activation process of the trehalose 6 phosphate synthase. PMID- 11429463 TI - The acid-stress response in Lactobacillus sanfranciscensis CB1. AB - Lactobacillus sanfranciscensis CB1, an important sourdough lactic acid bacterium, can withstand low pH after initial exposure to sublethal acidic conditions. The sensitivity to low pH varied according to the type of acid used. Treatment of LB: sanfranciscensis CB1 with chloramphenicol during acid adaptation almost completely eliminated the protective effect, suggesting that induction of protein synthesis was required for the acid-tolerance response. Two constitutively acid tolerant mutants, CB1-5R and CB1-7R, were isolated using natural selection techniques after sequential exposure to lactic acid (pH 3.2). Two-dimensional gel electrophoresis analysis of protein expression by non-adapted, acid-adapted and acid-tolerant mutant cells of LB: sanfranciscensis showed changes in the levels of 63 proteins. While some of the modifications were common to the acid-adapted and acid-tolerant mutant cells, several differences, especially regarding the induced proteins, were determined. The two mutants showed a very similar level of protein expression. Antibodies were used to identify heat-shock proteins DnaJ, DnaK, GroES and GrpE. Only GrpE showed an increased level of expression in the acid-adapted and acid-tolerant mutants as compared with non-adapted cells. The N terminal sequence was determined for two proteins, one induced in both the acid adapted and mutant cells and the other showing the highest induction factor of those proteins specifically induced in the acid-adapted cells. This second protein has 60% identity with the N-terminal portion of YhaH, a transmembrane protein of Bacillus subtilis, which has 54 and 47% homology with stress proteins identified in Listeria monocytogenes and Bacillus halodurans. The constitutively acid-tolerant mutants showed other different phenotypic features compared to the parental strain: (i) the aminopeptidase activity of CB1-5R decreased and that of CB1-7R markedly increased, especially in acid conditions; (ii) the growth in culture medium at 10 degrees C and in the presence of 5% NaCl was greater (the same was found for acid-adapted cells); and (iii) the acidification rate during sourdough fermentation in acid conditions was faster and greater. PMID- 11429464 TI - Characterization of an autostimulatory substance produced by Escherichia coli. AB - The recovery of dilute populations of stationary phase cells of Escherichia coli was studied using an automatic growth analyser. The addition of 30% supernatant from 2-d-old stationary phase cells of the organism reproducibly shortened the apparent lag times by 22-57.5%, depending on the age of the inoculum. True lag times, as determined by colony counts, of stationary phase cells were reduced by supernatant addition by 41-62%. The growth-stimulating substance was characterized and partly purified from supernatants: the active material was shown to be dialysable, heat-stable, acid- and alkali-stable and protease resistant. Extraction with ethyl acetate or ion-exchange resins was not successful, but the active material could be quantitatively extracted with ethanol after saturation with salt. It is concluded that the active substance is a small, non-proteinaceous, non-ionic organic molecule. Separation of extracts by HPLC indicated that the stimulatory substance is weakly hydrophobic and has retention times similar to those of uracil. So far, however, the exact chemical identity of the active substance has not been elucidated. PMID- 11429465 TI - The cryptic ushA gene (ushA(c)) in natural isolates of Salmonella enterica (serotype Typhimurium) has been inactivated by a single missense mutation. AB - Two mutational mechanisms, both supported by experimental studies, have been proposed for the evolution of new or improved enzyme specificities in bacteria. One mechanism involves point mutation(s) in a gene conferring novel substrate specificity with partial or complete loss of the original (wild-type) activity of the encoded product. The second mechanism involves gene duplication followed by silencing (inactivation) of one of these duplicates. Some of these 'silent genes' may still be transcribed and translated but produce greatly reduced levels of functional protein; gene silencing, in this context, is distinct from the more common associations with bacterial partitioning sequences, and with genes which are no longer transcribed or translated. Whereas most Salmonella enterica strains are ushA(+), encoding an active 5'-nucleotidase (UDP-sugar hydrolase), some natural isolates, including most genetically related strains of serotype Typhimurium, have an ushA allele (designated ushA(c)) which produces a protein with, comparatively, very low 5'-nucleotidase activity. Previous sequence analysis of cloned ushA(c) and ushA(+) genes from serotype Typhimurium strain LT2 and Escherichia coli, respectively, did not reveal any changes which might account for the significantly different 5'-nucleotidase activities. The mechanism responsible for this reduced activity of UshA(c) has hitherto not been known. Sequence analysis of Salmonella ushA(+) and ushA(c) alleles indicated that the relative inactivity of UshA(c) may be due to one, or more, of four amino acid substitutions. One of these changes (S139Y) is in a sequence motif that is conserved in 5'-nucleotidases across a range of diverse prokaryotic and eukaryotic species. Site-directed mutagenesis confirmed that a Tyr substitution of Ser-139 in Salmonella UshA(+) was solely responsible for loss of 5' nucleotidase activity. It is concluded that the corresponding single missense mutation is the cause of the UshA(c) phenotype. This is the first reported instance of gene inactivation in natural isolates of bacteria via a missense mutation. These results support a model of evolution of new enzymes involving a 'silent gene' which produces an inactive, or relatively inactive, product, and are also consistent with the evolution of a novel, but unknown, enzyme specificity by a single amino acid change. PMID- 11429466 TI - A chromosomal region surrounding the ompD porin gene marks a genetic difference between Salmonella typhi and the majority of Salmonella serovars. AB - In this work it is shown that the majority of Salmonella serovars most frequently associated with the systemic infection of vertebrate hosts produce a major outer membrane porin, OmpD. However, OmpD is absent from the outer-membrane protein profiles of Salmonella typhi strain Ty2 and 26 clinical isolates of S. typhi examined by SDS-PAGE. To determine whether the ompD gene is present in S. typhi, primers internal to the ompD coding sequence were used to amplify the gene by PCR. With the exception of S. typhi strains, the ompD gene was amplified from the genomes of all Salmonella serovars tested. Consistently, a specific ompD probe did not hybridize with DNA isolated from the S. typhi strains. Taken together, these results demonstrate that S. typhi does not produce OmpD due to the absence of the ompD gene. Furthermore, it was investigated whether the deletion of ompD extended to smvA. This gene is adjacent to ompD in the Salmonella typhimurium chromosome and encodes a protein involved in the resistance to methyl viologen, a superoxide-generating agent. Although PCR failed to amplify the smvA gene from the S. typhi strain Ty2 genome, it was possible to amplify it from the chromosome of the clinical strains. On the other hand, hybridization analyses showed that the smvA gene is present in all the S. typhi strains tested. In contrast to the other Salmonella serovars, S. typhi strain Ty2 and the clinical isolates showed sensitivity to methyl viologen, suggesting that smvA gene is inactive in S. typhi. In conclusion, the ompD-smvA region is variable in structure among Salmonella serovars. It is hypothesized that the absence of ompD may suggest a role in host specificity. PMID- 11429467 TI - Physical and genetic map of the Clostridium saccharobutylicum (formerly Clostridium acetobutylicum) NCP 262 chromosome. AB - A physical and genetic map of the Clostridium saccharobutylicum NCP 262 chromosome was constructed. The order of macrorestriction fragments was determined by analysing fragments generated after single and double digestion with the restriction enzymes BssHII, I-CeuI, Sse8387I, RsrII and SfiI and separation by PFGE. The I-CeuI backbone of C. saccharobutylicum was constructed by indirect end-labelling with rrs- and 3' rrl-specific probes located on either side of the I-CeuI site in the rrn operon, and reciprocal separation of BssHII and I-CeuI digestion products by two-dimensional PFGE. The positions of BssHII fragments on the physical map were determined using a library of linking clones containing BssHII cleavage sites. The size of the circular genome was estimated to be 5.3 Mb with a mean resolution of approximately 140 kb. The chromosome of C. saccharobutylicum contains 12 rrn operons, located on 46% of the chromosome, which are transcribed divergently from the deduced origin of replication. The genetic map was constructed by determining the location of 28 genes involved in house-keeping, heat-shock response, sporulation, electron transfer and acid- and solvent-formation. Comparison of the C. saccharobutylicum genetic map with those of the spore-forming bacteria Bacillus subtilis, Clostridium acetobutylicum, Clostridium perfringens and Clostridium beijerinckii indicated C. saccharobutylicum to be most similar to the latter two Clostridium species, with the order of the genes within the gyrAB and recA loci being conserved. PMID- 11429468 TI - The double mechanism of incompatibility between lambda plasmids and Escherichia coli dnaA(ts) host cells. AB - For plasmids derived from bacteriophage lambda, the initiation of bidirectional DNA replication from orilambda depends on the stimulation of transcription from the p(R) promoter by the host replication initiator protein DnaA. Certain Escherichia coli dnaA(ts) mutants cannot be transformed by wild-type lambda plasmids even at the temperature permissive to cell growth. This plasmid-host incompatibility appeared to be due to inefficient stimulation of transcription from the p(R) promoter by the mutant DnaA protein. This paper shows that there is a second mechanism for the incompatibility between lambda plasmids and dnaA(ts) hosts, exemplified in this study by the dnaA46 mutant. This is based on the competition between the lambda P protein and the host DnaA and DnaC proteins for DnaB helicase. Both mechanisms must be operative for the incompatibility. PMID- 11429469 TI - Mosaic structure of Shiga-toxin-2-encoding phages isolated from Escherichia coli O157:H7 indicates frequent gene exchange between lambdoid phage genomes. AB - Shiga-toxin-2 (stx(2))-encoding bacteriophages were isolated from Norwegian Escherichia coli O157:H7 isolates of cattle and human origin. The phages were characterized by restriction enzyme analysis, hybridization with probes for toxin genes and selected phage DNA such as the P gene, integrase gene and IS1203, and by PCR studies and partial sequencing of selected DNA regions in the integrase to stx(2) region of the phages. The stx(2)-phage-containing bacteria from which inducible phages were detected produced similar amounts of toxin, as shown by a Vero cell assay. The results indicate that the population of stx(2)-carrying phages is heterogeneous, although the phages from epidemiologically linked E. coli O157:H7 isolates were similar. There appears to have been frequent recombination of stx(2) phages with other lambdoid phages. PMID- 11429470 TI - Analysis of the wee gene cluster responsible for the biosynthesis of the polymeric bioemulsifier from the oil-degrading strain Acinetobacter lwoffii RAG 1. AB - A cluster (27 kbp) of genes responsible for the biosynthesis of the amphipathic, polysaccharide bioemulsifier emulsan from the oil-degrading Acinetobacter lwoffii RAG-1 was isolated and characterized. The complete sequence of this cluster, termed wee, consisted of 20 ORFs. One set of 17 ORFs was transcribed in one direction, while a second set of three ORFs, 607 bp upstream of the first, was transcribed in the opposite direction. Mutations in either of the two regions caused defects in emulsan production, yielding specific activities of 5-14% of parental emulsifying activity. Putative functions could be assigned to proteins involved in production of nucleotide amino sugar precursors, transglycosylation, transacetylation, polymerization and transport. However, no JUMPstart or ops sequences, normally found associated with some polysaccharide biosynthetic gene clusters, were identified. Evidence is presented suggesting that the bioemulsifier may be a member of the group 1 or group 4 polysaccharides. PMID- 11429471 TI - Ralstonia metallidurans CH34 RpoN sigma factor and the control of nitrogen metabolism and biphenyl utilization. AB - Ralstonia metallidurans CH34 can use biphenyl as carbon and energy source when provided with the catabolic transposon Tn4371. Previous results suggested that this property was dependent on the RNA polymerase subunit sigma(54). The authors sequenced the CH34 rpoN gene and flanking DNA and isolated a CH34 rpoN-deficient strain. Analysis of the sequence revealed a set of features conserved in all rpoN genes and flanking DNA regions previously analysed in other bacterial species. Nevertheless, despite this conservation, CH34 differed even from the closely related strain R. eutropha H16 by one particular ORF. The rpoN null mutation did not affect expression of the Tn4371 bph operon although it did alter the ability of the Tn4371 host strain to grow on biphenyl. The CH34 rpoN mutant had lost the capacity for autotrophic growth and for responding to poor nitrogen sources by a decrease in urease and proline oxidase activity. CH34 RNA polymerase sigma(54) thus positively controls autotrophy as well as nitrogen metabolism but only indirectly affects Tn4371-directed biphenyl utilization. PMID- 11429472 TI - A diffusible analogue of N(3)-(4-methoxyfumaroyl)-L-2,3-diaminopropanoic acid with antifungal activity. AB - N(3)-(4-Methoxyfumaroyl)-L-2,3-diaminopropanoic acid (FMDP), a specific and potent inactivator of glucosamine-6-phosphate (GlcN-6-P) synthase from Candida albicans, exhibits relatively poor anticandidal activity, with an MIC value amounting to 50 microg ml(-1) (200 microM). Uptake of FMDP into C. albicans cells follows saturation kinetics and is sensitive to the action of metabolic inhibitors, thus indicating the active transport mechanism. However, the acetoxymethyl ester of FMDP penetrates the fungal cell membrane by free diffusion and is rapidly hydrolysed by C. albicans cytoplasmic enzymes to release the free FMDP. This mechanism gives rise to continuous accumulation of the enzyme inhibitor and results in higher antifungal activity of the FMDP ester (MIC=3.1 microg ml(-1), 10 microM). These results show that the 'pro-drug' approach can be successfully applied for the enhancement of antifungal activity of glutamine analogues that inhibit GlcN-6-P synthase. PMID- 11429473 TI - Overexpression of a dominant-negative allele of YPT1 inhibits growth and aspartyl protease secretion in Candida albicans. AB - To investigate the pre-Golgi secretion pathway in the pathogenic yeast Candida albicans, we cloned the C. albicans homologue of the Saccharomyces cerevisiae protein secretion gene YPT1. The C. albicans YPT1 ORF contained a 624 bp intronless ORF encoding a deduced protein of 207 aa and 2.3 kDa. This deduced protein was 77% identical to S. cerevisiae Ypt1 protein (Ypt1p) and it contained GTP-binding domains that are conserved among all known ras-like GTPases. Multicopy plasmids containing C. albicans YPT1 complemented the temperature sensitive S. cerevisiae ypt1 (A136D) mutation. One chromosomal YPT1 allele in C. albicans CAI4 was readily disrupted by homologous gene targeting, but attempts to disrupt the second allele yielded no viable null mutants. Since this suggested that C. albicans YPT1 may be essential, a mutant ypt1 allele was constructed encoding the amino acid substitution analogous to the N121I substitution in a known trans-dominant inhibitor of S. cerevisiae Ypt1p. Next, a GAL1-regulated plasmid was used to express the mutant ypt1(N121I) allele in C. albicans CAI4. Ten of 11 transformants tested grew normally in glucose and poorly in galactose, and plasmid curing restored growth to wild-type levels. When these transformants were incubated in galactose, secretion of aspartyl proteinase (Sap) was inhibited and membrane-bound secretory vesicles accumulated intracellularly. These results imply that C. albicans YPT1 is required for growth and protein secretion, and they confirm the feasibility of using inducible dominant-negative alleles to define the functions of essential genes in C. albicans. PMID- 11429474 TI - Different adhesins for type IV collagen on Candida albicans: identification of a lectin-like adhesin recognizing the 7S(IV) domain. AB - Adherence of the opportunistic pathogen Candida albicans to basement membrane (BM) proteins is considered a crucial step in the development of candidiasis. In this study the interactions of C. albicans yeast cells with the three main domains of type IV collagen, a major BM glycoprotein, were analysed. C. albicans adhered to the three immobilized domains by different mechanisms. Adhesion to the N-terminal cross-linking domain (7S) required the presence of divalent cations, whereas interaction with the central collagenous domain (CC) was cation independent. Recognition of the C-terminal non-collagenous domain (NC1) was partially cation-dependent. Binding inhibition assays with the corresponding domains in soluble form showed that these interactions were specific. Both Ca(2+) and Mg(2+) promoted adhesion to the 7S domain and the interaction was completely abolished by EDTA. Treatment of the 7S domain, or its subunits, with N glycosidase F reduced yeast binding by approximately 70%. Moreover, several sugars known to be part of the N-linked oligosaccharide chains of collagen IV inhibited adhesion to immobilized 7S; N-acetylglucosamine, L-fucose and methylmannoside caused a similar inhibition whereas N-acetyllactosamine was a more effective inhibitor. In contrast, glucose, galactose, lactose or heparan sulfate did not affect yeast binding. Combinations of the inhibitory sugars at suboptimal inhibition concentrations did not reduce C. albicans adhesion more than the individual sugars, pointing to a single lectin as responsible for the interaction. These results taken together show that C. albicans utilizes several adhesins for interacting with type IV collagen, and that at least one of them is a lectin which recognizes the 7S(IV) oligosaccharide residues as its receptor. PMID- 11429475 TI - Secretion, interaction and assembly of two O-glycosylated cell wall antigens from Candida albicans. AB - The mechanisms of incorporation of two antigens have been determined using a monoclonal antibody (3A10) raised against the material released from the mycelial cell wall by zymolyase digestion and retained on a concanavalin A column. One of the hybridomas secreted an IgG that reacted with two bands in Western blots. Indirect immunofluorescence showed that the antigens were located on the surfaces of mycelial cells, but within the cell walls of yeasts. These antigens were detected in a membrane preparation, in the SDS-soluble material and in the material released by a 1,3-beta-glucanase and chitinase from the cell walls of yeast and mycelial cells. In the latter three samples, an additional high molecular-mass, highly polydispersed band was also detected. Beta-elimination of each fraction resulted in the disappearance of all antigen bands, suggesting that they are highly O-glycosylated. In addition, the electrophoretic mobility of the high-molecular-mass, highly polydispersed bands increased after digestion with endoglycosidase H, indicating that they are also N-glycosylated. New antigen bands were released when remnants of the cell walls extracted with 1,3-beta glucanase or chitinase were digested with chitinase or 1,3-beta-glucanase. These results are consistent with the notion that, after secretion, parts of the O glycosylated antigen molecules are transferred to an N-glycosylated protein(s). This molecular complex, as well as the remaining original 70 and 80 kDa antigen molecules, next bind to 1,3-beta-glucan or chitin, probably via 1,6-beta-glucan, and, in an additional step, to chitin or 1,3-beta-glucan. This process results in the final molecular product of each antigen, and their distribution in the cell walls. PMID- 11429476 TI - Respiratory infectious complications during chemotherapy-induced aplasia. PMID- 11429477 TI - The spectrum of pulmonary infections in cancer patients. AB - The lung is a common site of infection in patients with cancer. The spectrum of pulmonary infection depends on the underlying immunologic deficit or deficits. In neutropenic patients, gram-negative bacterial infections predominate early, whereas fungal infections (Aspergillus, Zygomycetes, Fusarium species) are common if neutropenia persists. In patients with impaired cellular immunity, viral infections (cytomegalovirus, other herpes viruses) predominate and may coexist with bacterial (Legionella, Nocardia), mycobacterial, and fungal (Aspergillus, Histoplasma, etc.) infections. Pneumocystis carinii pneumonia is also common in this setting. Infections caused by Streptococcus pneumoniae and Haemophilus influenzae are the primary bacterial infections encountered in patients with impaired humoral immunity. In patients with primary or metastatic pulmonary neoplasms, postobstructive pneumonitis, lung abscess, and occasionally empyema of mixed bacterial etiology (Staphylococcus species, gram-negative bacilli, anaerobes) are frequent. Patients with brain tumors and head and neck cancer develop aspiration pneumonitis, which is usually caused by organisms living in the oropharynx and upper airways. Several immunologic deficits might be present in the same patient, making such a patient susceptible to a wide variety of opportunistic pathogens. PMID- 11429478 TI - Mechanical ventilation in critically ill cancer patients. AB - Outcomes for cancer patients undergoing mechanical ventilation have improved in recent times, and the outlook for this patient group is no longer hopeless. Advances in ventilation techniques and increasingly sophisticated equipment have allowed oncologic intensivists to intervene more effectively in their patients' critical illness. In addition, many of these patients are now referred for intensive care much earlier. This article reviews the use of mechanical ventilation in critically ill cancer patients and includes discussion of the recently published outcome literature. We also discuss some recent advances in the broader field of mechanical ventilation and demonstrate how these advances can be applied to the oncology population. PMID- 11429479 TI - Pneumonia in febrile neutropenic patients: radiologic diagnosis. AB - Pneumonia in febrile neutropenic cancer patients should be diagnosed as early as possible, because prompt institution of targeted therapeutic measures might be essential for their prognosis. Conventional chest radiographs frequently fail to detect lung infiltrates at an early stage, meaning that a normal chest radiograph finding must be interpreted with caution. Thoracic computed tomograph scans provide a much higher yield and are therefore recommended in patients at risk for a complicated pulmonary infection. Lung infiltrates documented by computed tomograph scans (eg, nodular infiltrates with or without a halo, ground-glass opacities, or cavitations with or without air crescent signs) open up a wide range of differential diagnoses, such as invasive pulmonary aspergillosis, other types of pneumonia, hemorrhage, infiltration by the underlying malignancy, drug toxicity, alveolar proteinosis, or acute respiratory distress syndrome. High resolution techniques or magnetic resonance imaging may provide further details to help distinguish inflammatory processes from processes that may not require an antimicrobial intervention. Sequential nonculture-based monitoring for invasive fungal infections, using Aspergillus antigen sandwich enzyme-linked immunosorbent assay, and panfungal or Aspergillus-specific polymerase chain reaction, may add important tools in the early identification of patients who may benefit from systemic antifungal treatment. PMID- 11429480 TI - Bronchoscopic invasive diagnostic techniques in the cancer patient. AB - The cancer patient often presents with fever and pulmonary infiltrates, in particular during the course of chemotherapy or after bone marrow transplantation. In these conditions, specific diagnoses are mainly related to an infective cause, but noninfectious processes, malignant or not, are also found alone or in combination with infection. Identification of the pulmonary process can be achieved by bronchoscopic techniques, including bronchoalveolar lavage (BAL) and transbronchial biopsy (TBB). BAL may help identify opportunistic organisms but also bacterial pneumonia, provided quantitative cultures are performed, and TBB has been shown to increase the diagnostic yield of BAL. These two procedures should then be combined, provided there is no contraindication. PMID- 11429481 TI - Radiation-induced and chemotherapy-induced pulmonary injury. AB - The management of cancer has continued to advance with the development of new chemotherapeutic agents and improved techniques of radiation therapy. Although new therapeutic approaches have improved survival in cancer patients, each form of intervention has the potential to produce adverse effects on normal host tissues. Some of these toxicities may be accentuated with combined modality therapy. The use of chemotherapy and radiation therapy, alone or combined, can be associated with clinically significant pulmonary toxicity. The pulmonary toxic effects of chemotherapy can be divided into (1) early onset, resulting in interstitial lung injury, and (2) late onset, with pulmonary fibrosis as a sequela. These toxic effects are frequently dose related but may be enhanced by radiation therapy. Similar to chemotherapy, radiation can produce acute or chronic lung injury depending on dose rate, duration, preexisting lung disease, and concomitant steroid use. Acute radiation injury typically occurs 2 weeks to 3 months after treatment and is usually limited to the irradiated field. Mild injury often resolves without treatment, whereas more serious injury results in fibrosis 6 to 12 months after treatment. Histopathologic evaluation of acute lung injury is no different from drug-induced injury, and damage to vascular endothelial cells and alveolar lining cells is seen. This article reviews and provides an update on the clinically important chemotherapy and radiation-induced pulmonary injuries, the pathologic mechanisms, where known, and the treatment advances that have occurred in this field. PMID- 11429482 TI - Signal transduction pathways in sarcoma as targets for therapeutic intervention. AB - Investigations into the molecular alterations in sarcomas have made substantial progress during the past decade. Classical linkage analysis and the direct sequencing of chromosomal translocation fusions have identified candidate genes in many different sarcomas. A large group of these genes participate in signal transduction pathways and represent potential sites of disease intervention with targeted therapies. This review will discuss five types of sarcoma that display aberrant tyrosine kinase pathway signaling: gastrointestinal stromal tumor, inflammatory myofibroblastic tumor, congenital fibrosarcoma and mesoblastic nephroma, dermatofibrosarcoma protuberans, and desmoplastic small round cell tumor; one sarcoma predisposition syndrome with specific dysregulation of the ras pathway--neurofibromatosis--will also be discussed. PMID- 11429483 TI - Recent progress in the classification of soft tissue tumors: role of genetics and clinical implications. AB - Soft tissue tumors comprise a vast and heterogeneous group of neoplasms. Because different tumors often have different biological behaviors and respond differently to various therapeutic modalities, precise classification is paramount. The majority of soft tissue tumors were first delineated on the basis of morphologic and clinical findings, which in many cases were adequate to accurately separate different tumors into homogeneous groups; however, it has increasingly been appreciated that many entities are actually heterogeneous groups of tumors that have similar histologic and pathologic characteristics but differ in their clinical behavior and underlying pathogenesis. Within the past several years, great strides have been made in the purification of different entities. This accomplishment has largely been because of advances in our understanding of the molecular genetics that underlie the pathogenesis of many sarcomas and the development of new and specific tumor markers. This review highlights some important recent work in two selected soft tissue tumors gastrointestinal stromal tumor and inflammatory myofibroblastic tumor. These examples illustrate the type of progress that is being made in the classification of soft tissue tumors. PMID- 11429484 TI - Angiogenesis and antiangiogenic approaches to sarcomas. AB - Established chemotherapy regimens for advanced or metastatic sarcoma generally have low response rates and substantial toxicity, and resistance often arises quickly, making sarcoma an ideal target for alternative treatment approaches. Antiangiogenic therapies have a number of potential advantages including decreased resistance, fewer side effects, and a broad spectrum of activity. This paper reviews the evidence supporting the use of antiangiogenic therapies for adult soft tissue sarcomas. Human sarcomas express a number of proangiogenic factors that may represent therapeutic targets, with vascular endothelial growth factor being the best characterized, and inhibitors of angiogenesis have antitumor activity in animal models of human sarcomas. Clinical trials are in early stages, although promising results have already been seen. In the future, improved drugs, refined molecular profiling of tumors, and new ways of combining antiangiogenic agents with cytotoxic agents may lead to more effective and tolerable therapies for sarcomas. PMID- 11429485 TI - Soft tissue sarcoma as a model disease to examine cancer immunotherapy. AB - Immunotherapy is in its infancy for many diseases, whether they are neoplastic or autoimmune. The major issues for cancer immunotherapy today involve the definition of molecular targets and the generation of effector mechanisms to attack the targets of interest. Soft tissue sarcomas provide a unique opportunity to examine the immune response against defined antigens. Many types of sarcomas contain tumor-specific chromosomal translocations encoding fusion proteins, which are attractive targets for immunotherapy. Our understanding of the immune system is also coming into clearer focus with the discovery of dendritic cells as powerful natural adjuvants and the teasing out of mechanisms leading to immunity versus tolerance as examples. It is hoped that the intersection of cellular immunology and sarcoma molecular biology will lead to new modalities of therapy for this group of patients with this heterogeneous group of diseases. PMID- 11429486 TI - New chemotherapy approaches in colorectal cancer. AB - Colorectal cancer is one of the leading causes of cancer deaths in the Western world, with approximately 50% of all patients dying from metastatic disease. Until recently, therapeutic options for advanced colorectal cancer were mainly confined to chemotherapy with 5-fluorouracil in various schedules, with or without biochemical modulation with leucovorin. The development of new cytotoxic drugs with substantial activity in this tumor during the past 2 years has dramatically changed treatment strategies and therapeutic goals in metastatic colorectal cancer and has introduced neoadjuvant chemotherapy followed by secondary surgery with the intent of long-term survival. Among these new drugs, oral fluoropyrimidines (tegafur/uracil and capecitabine), irinotecan, and oxaliplatin have already established themselves as part of various treatment approaches. Other novel therapeutics including agents designed to act on molecular targets already show promising activity and will become part of combination protocols with current standard chemotherapy. PMID- 11429487 TI - Adjuvant therapy for noncolorectal cancers. AB - Cancers of the esophagus, stomach, and pancreas account for 11% of American cancer deaths and have a high case fatality rate. For esophageal cancer, the superiority of chemoradiotherapy rather than radiotherapy alone as a nonsurgical management was reaffirmed by a large pattern-of-care study in the United States. The study of preoperative chemoradiotherapy followed by surgery continues, with the investigation of newer chemotherapeutic agents combined with radiotherapy in an attempt to improve the therapeutic index of therapy. Trials attempting to intensify chemoradiotherapy treatments have included the addition of postoperative chemotherapy, the addition of brachytherapy, and the escalation of radiotherapy dose above the standard dose of 50.4 Gy. Neither brachytherapy nor an increase in external beam radiotherapy dose has been proven to improve local tumor control or patient survival. Adjuvant chemotherapy alone may have an impact on patient survival in one preliminary report, despite the results of earlier trials that failed to show a benefit for adjuvant chemotherapy alone. In the adjuvant treatment of gastric cancer, a meta-analysis of adjuvant chemotherapy trials suggested a survival benefit for adjuvant chemotherapy compared with surgery alone; however, preliminary reports of two large adjuvant chemotherapy trials using cisplatin-based chemotherapy failed to improve survival compared with surgery alone. The large Intergroup Trial 116, comparing surgery alone to surgery followed by postoperative fluorouracil, leucovorin, and radiotherapy, indicated a significant survival benefit for postoperative chemoradiotherapy. Postoperative chemoradiotherapy is the new standard of care for high-risk resected gastric cancer. Ongoing and future trials will address the inclusion of newer chemotherapeutic agents, the use of preoperative chemotherapy and radiotherapy, and the use of intraperitoneal therapy. In the adjuvant treatment of pancreatic cancer, clinical trials continue to evaluate the role of fluorouracil and radiotherapy, the use of preoperative chemoradiotherapy, and the incorporation of new therapeutic agents. PMID- 11429488 TI - Predictive and prognostic markers in gastrointestinal cancers. AB - As the roles of specific genes and their functions in cancer etiology and progression become defined, the necessity of looking at multiple genes and their interaction becomes clearer. In gastrointestinal cancer, tumor suppressor and promoter genes, cell cycle control and angiogenic factors, repair genes, and metabolic genes have been examined as they relate to prognosis and therapeutic outcomes. To date, most correlative studies for specific molecular parameters have been done retrospectively. Methodologies, sample sizes, and definitions differ. The following analysis provides a brief review of the current state of knowledge and recent research on the most commonly studied molecular markers, and the newer areas of inquiry attempting to define how to use molecular-genetic features of individual tumors as targets for more effective therapies. Consideration is given to the design of trials employing molecular targets as outcome factors. PMID- 11429489 TI - Radiation therapy in the management of rectal cancer. AB - Substantial advances have been made in the adjuvant management of patients with resectable rectal cancer. Increasing interest in patient quality of life has promoted the use of radiation therapy to enhance sphincter-preserving surgical approaches as an alternative to the standard abdominoperineal resection. Because of the suggestion of enhanced sphincter preservation with preoperative therapy and the potential advantage of decreased acute morbidity, randomized trials comparing preoperative and postoperative adjuvant combined modality therapy are ongoing. Recent progress in adjuvant postoperative treatment regimens relates to the integration of systemic therapy to radiation, and redefining the techniques for both modalities. The incorporation of improved radiation planning may reduce treatment-related bowel toxicity. The integration of novel chemotherapeutic agents in the adjuvant therapy of rectal cancer remains an active area of investigation. PMID- 11429491 TI - Evolving concepts in surgical management of urinary calculi. PMID- 11429490 TI - Chemoprevention of colorectal cancer. AB - Chemopreventive strategies hold substantial promise for reducing the incidence of colorectal cancer, the second leading cause of cancer-related mortality in the United States. This review focuses on recent advances in the identification of molecular targets and novel strategies for chemopreventive intervention. Many clinical trials are now in progress to assess the ability of synthetic agents or nutritional supplements to alter either the number of colorectal adenomas or biomarkers associated with colorectal tumorigenesis. Populations under study include genetically defined high-risk people and those with increased risk based on a personal history of colorectal neoplasia. A recent study showing that celecoxib, a cyclooxygenase-2 inhibitor, can alter the natural history of polyp formation in patients with familial adenomatous polyposis has provided a benchmark for the clinical development of other chemopreventive agents and heightened awareness that colorectal cancer is a preventable disease. PMID- 11429492 TI - Metabolic evaluation of stone disease patients: a practical approach. AB - The high success rates of extracorporeal shockwave lithotripsy and endourological techniques, together with their ease of use, in the treatment of urinary stones often overshadow the importance of the metabolic component of stone disease. It is my opinion that the prevention of further stones complements management of the acute episode. This review summarizes a variety of approaches to the measurement and manipulation of individual risk factors in recurrent urinary stone disease. PMID- 11429493 TI - Medical and surgical therapy of the cystine stone patient. AB - Cystinuria is an inherited defect in renal tubular and intestinal transport of dibasic amino acids that results in elevated urinary excretion of cystine, ornithine, lysine, and arginine. The only clinical manifestation of this disease is the development of urinary tract cystine calculi. Cystinuric patients suffer recurrent stone episodes, requiring an aggressive multi-modal approach to management. This article reviews the results of medical prevention regimens, the role of minimally invasive urologic intervention, and the recent insights into the complex genetic basis for the varied cystinuric phenotypes. PMID- 11429494 TI - Renal collecting system anatomy: its possible role in the effectiveness of renal stone treatment. AB - The anatomy of the kidney collecting system may play a role in the selection of the best method of kidney stone treatment for a specific patient. Also, an analysis of the collecting-system anatomy would indicate the likely effectiveness of each method of treatment. For stones located in the lower pole, the clearance rate after shockwave lithotripsy has been uniformly low relative to that of calculi elsewhere. Some special anatomical findings suggest that retention of what are considered to be 'passable stone fragments' (4 mm in diameter or less) in the inferior pole might be a consequence not only of the gravity-dependent position of lower calices but also of particular anatomical features of the inferior pole collecting system. The aspects reviewed and discussed are the angle formed between the main lower infundibulum and the renal pelvis (the infundibulopelvic angle), the lower infundibula diameters, the lower infundibulum length and height, and the lower calices spatial distribution. Also, the presence of minor calices perpendicular to the surface of the collecting system and drainage of superior and inferior poles are reviewed and discussed in the context of their importance to the effectiveness of renal stone treatment. PMID- 11429495 TI - An update on percutaneous nephrolithotomy in the management of urinary calculi. AB - Since its introduction in the 1970s, percutaneous nephrolithotomy has undergone numerous refinements in both instrumentation and technique to become an efficacious and safe procedure for the removal of renal calculi. The aim of this review is to discuss issues raised in publications in the year 2000 regarding percutaneous nephrolithotomy in the management of urinary stone disease. PMID- 11429496 TI - New concepts in the treatment of ureteral calculi. AB - The management of patients with ureteral stones remains under debate in several areas. The ability to predict spontaneous passage has improved but remains imprecise, whilst the range of therapeutic options continues to widen. Excellent results can be obtained by both shockwave lithotripsy and ureteroscopic methods, with relatively minimal complications. Routine ureteral stenting is not warranted whichever treatment is chosen. In future, directly comparative studies should be designed to incorporate quality-of-life parameters rather than just stone-free status, to improve our understanding of the effect of treatment decisions on patients. PMID- 11429497 TI - The management of urinary calculi in pregnancy. AB - When a pregnant patient presents with a urinary calculus, the customary investigations and management must change in order to take into account the well being of the developing fetus. Transabdominal or endovaginal ultrasound should be the initial imaging modality used in order to establish the diagnosis. A plain abdominal X-ray, limited intravenous pyelography, or retrograde pyelography is used secondarily if a definitive diagnosis is lacking. The treatment of first choice for urolithiasis in pregnancy is conservative, because 70-80% of stones will pass spontaneously. If conservative management fails, or in cases of sepsis, obstruction of a solitary kidney, or bilateral ureteric obstruction, then surgical intervention is indicated. Traditional surgical management consists of draining the obstructed collecting system with a ureteral stent or percutaneous nephrostomy tube with definitive treatment of the stone in the post-partum period. Ureteroscopic lithotripsy and stone extraction is another option that has been used safely and reliably with increasing frequency in many centers. Despite recent reports of using extracorporeal shock-wave lithotripsy, this treatment is still considered contraindicated in pregnancy. PMID- 11429498 TI - Pediatric urolithiasis. AB - Urinary lithiasis in the pediatric population has evolved from a mere clinical curiosity to a disease process worthy of thoughtful and rigorous scientific study. All aspects of urinary lithiasis have undergone this evolution, including the epidemiology of stone formation in children, defining new modalities of radiologic imaging, and refining surgical techniques for stone treatment. These advancements and observations in pediatric stone disease are highlighted. PMID- 11429499 TI - Economics of urolithiasis: cost-effectiveness of therapies. AB - The cost of treating urolithiasis with extracorporeal shockwave lithotripsy and endoscopic surgery continues to be a significant burden on a nation's healthcare economy. Cost-effectiveness evaluations of various medical and surgical treatment options for urolithiasis is a practical method of developing rational allocation strategies for limited economic resources. In this review, the cost-effectiveness of shockwave lithotripsy and that of endoscopic surgery in the management of kidney and ureteral stones are compared. This is followed by a discussion of the cost of the medical management of urolithiasis in comparison with repeated surgical treatment. PMID- 11429500 TI - Voiding dysfunction: definitions. AB - 'Lower urinary tract symptoms' is a term that describes symptoms related to both the storage and emptying phases of the micturition cycle. Storage symptoms include urinary frequency urgency, urge incontinence, nocturia, dysuria and other kinds of pain emanating from the bladder or urethra. Emptying symptoms consist of hesitancy, straining to void, difficulty starting, diminished stream, a feeling of incomplete bladder emptying and urinary retention. In both sexes, the etiology of lower urinary tract symptoms is multifactorial, and symptoms are a poor indicator of underlying pathophysiology. In men, lower urinary tract symptoms are most often attributed to prostatic obstruction, but only approximately two-thirds of men with lower urinary tract symptoms meet the accepted diagnostic criteria for obstruction. Approximately half have detrusor overactivity and a smaller number have impaired detrusor contractility, sensory urgency, sphincteric incontinence, polyuria or nocturnal polyuria. In women, lower urinary tract symptoms are often considered to result from hormonal abnormalities, childbirth, aging, or previous surgery, but the multifactorial underlying pathophysiology is similar to that seen in men, except for a much lower incidence of urethral obstruction and a high incidence of sphincteric incontinence. Treatment typically begins with empiric, conservative therapies aimed at resolving detrusor instability or bladder outlet obstruction. However, although either or both of these etiologies may exist in the individual with lower urinary tract symptoms, treatment may fail as a result of another cause. We believe that treatment based on the pathophysiology of the symptoms will lead to better outcomes than treatment based on symptoms alone. PMID- 11429501 TI - Sacral nerve neuromodulation in the treatment of refractory motor urge incontinence. AB - Sacral nerve neuromodulation may be an effective treatment option in patients with motor urge incontinence when conservative therapy fails. The effect of treatment is durable in the majority of patients, but there is also a high failure rate. The cause of failure remains unclear in most cases as it is still unknown exactly how neuromodulation works. The present review summarizes the results of patient treatment and research into the working mechanism of neuromodulation, as well as the technical developments described in the past year. PMID- 11429502 TI - Pubovaginal slings: a review of the technical variables. AB - The pubovaginal sling, reintroduced in the late 1970s by Maguire and Blaivas, has become the gold standard for managing anatomic incontinence. Newer technology, materials, surgical techniques and even new theories on the mechanism of action are evolving to further reduce the morbidity of these procedures and improve patient satisfaction. In the following review, we will highlight some of the exciting advances we have witnessed over the last year and try to put them into perspective for the reader. PMID- 11429503 TI - Post-urethral suspension obstruction. AB - Urethral obstruction is a consequence of surgery to treat stress incontinence in women. Although its incidence is relatively low, it still presents a challenging problem for the urologist. Recent work has focused on clarifying the etiology and incidence of this condition, as well as providing new definitions on bladder outlet obstruction in women. In addition, effective, less invasive ways of treating this condition have been described. PMID- 11429504 TI - Current management of vesicovaginal fistulae. AB - Vesicovaginal fistula continues to be a distressing problem that results most commonly from urogynecologic surgery. Several surgical techniques for correction of vesicovaginal fistulae have been described. The present review emphasizes those advances in surgical management and recent changes in etiology. PMID- 11429505 TI - Diagnostic imaging of pelvic floor dysfunction. AB - Pelvic floor dysfunction encompasses a variety of fascial and anatomic defects that can include a combination of cystocele, rectocele, uterine prolapse, enterocele and vault prolapse. Accurate diagnosis of the coexisting abnormalities is essential in planning reconstructive procedures so that the risks of recurrence and reoperation can be minimized. At this time, dynamic magnetic resonance imaging is the study of choice to evaluate the female pelvis and delineate the possible components of pelvic floor dysfunction. PMID- 11429506 TI - The use of synthetics in the treatment of pelvic organ prolapse. AB - Despite an improved understanding of pelvic anatomy and organ function and advances in surgical techniques, long-term success rates in pelvic surgery are still variable (3-59%), but can reach up to 92% in the case of associated procedures. The major causes of recurrent pelvic prolapse after corrective surgery are related to patient factors, such as poor tissues, impaired healing processes and chronic pathological increases in intra-abdominal pressure. Other causes of failure are, however, probably related to surgical techniques: the use of weak or insufficient sutures, or suboptimal performance of the surgery. In line with progress in the surgical correction of abdominal hernias, the use of synthetic mesh in pelvic reconstructive surgery also seems to guarantee its reliability, especially when autologous tissue is of poor quality or insufficient quantity. Moreover, the use of artificial mesh may simplify a surgical procedure, reducing operative time and the need for additional techniques. This review evaluates the main properties of synthetic biomaterials, their complications and the most common procedures involved in the use of synthetic mesh: the abdominal sacral colpopexy and transvaginal cystocele repair. PMID- 11429507 TI - Treatment of severe malnutrition. PMID- 11429508 TI - Protein-energy malnutrition: there is still work to do. PMID- 11429509 TI - Management of severe malnutrition: efficacious or effective? PMID- 11429510 TI - Measuring health-related quality of life of pediatric patients. PMID- 11429511 TI - Nitric oxide and intestinal water and electrolyte transport: in, out, or both? PMID- 11429512 TI - Proabsorptive action of gum arabic: regulation of nitric oxide metabolism in the basolateral potassium channel of the small intestine. AB - BACKGROUND: Gum arabic, a high-molecular-weight natural polysaccharide, has been shown to have proabsorptive properties in animal models of gastrointestinal disease that involve nitric oxide (NO). Gum arabic may indirectly regulate NO metabolism by creating an outward NO gradient, thus altering other intracellular NO-dependent mechanisms such as gating of the potassium (K+) channel. This hypothesis was further investigated using the K+ channel blocker, glybenclamide. METHODS: Following intraperitoneal injection of 4.5 mg/kg glybenclamide or saline, the jejunum of anesthetized rats was perfused with a standard oral rehydration solution in the presence or absence of 2.5 g/L gum arabic, as well as 1 mmol/L l-arginine to enhance NO production. Sodium, net water, and glucose absorption and unidirectional water movement were determined. RESULTS: Gum arabic showed regulatory capacity for NO-dependent metabolism by reducing net water absorption in the absence of arginine, and sodium absorption after arginine stimulation, in the absence of glybenclamide. Addition of gum arabic to oral rehydration solution, in glybenclamide pretreated animals, and in the absence of arginine, normalized sodium absorption, but was less effective in restoring net water transport. Injection of glybenclamide sharply decreased all absorption markers in arginine supplemented oral rehydration solution, which were at least partially restored by addition of gum arabic to the oral rehydration solution. In the presence of glybenclamide, the effects of arginine became antiabsorptive, as had those observed in preceding studies with high arginine concentration. Gum arabic partially or fully reversed alterations produced by perfused 1 mmol/L arginine. CONCLUSIONS: Some of the effects of gum arabic on the small intestine are likely caused by its ability to remove NO as it diffuses into the lumen, thus reducing NO concentration in the enterocyte and indirectly affecting the absorptive/secretory response of the gut, which leads to normalization of absorptive function. These findings are consistent with the previously shown gum arabic-scavenging properties of NO and support a potential therapeutic role for this product. PMID- 11429513 TI - Gas production by feces of infants. AB - BACKGROUND: Intestinal gas is thought to be the cause abdominal discomfort in infants. Little is known about the type and amount of gas produced by the infant's colonic microflora and whether diet influences gas formation. METHODS: Fresh stool specimens were collected from 10 breast-fed infants, 5 infants fed a soy-based formula, and 3 infants fed a milk-based formula at approximately 1, 2, and 3 months of age. Feces were incubated anaerobically for 4 hours at 37 degrees C followed by quantitation of hydrogen (H2), methane (CH4), carbon dioxide (CO2), hydrogen sulfide (H2S), methanethiol (CH3SH), and dimethyl sulfide (CH3SCH3) in the head-space. RESULTS: H2 was produced in greater amounts by breast-fed infants than by infants in either formula group, presumably the consequence of incomplete absorption of breast milk oligosaccharides. CH4 was produced in greater amounts by infants fed soy formula than by infants on other diets. CO2 was produced in similar amounts by infants in all feeding groups. Production of CH3SH was conspicuously low by feces of breast-fed infants and production of H2S was high by soy-formula-fed infants. CH3SCH3 was not detected. Only modest changes with age were observed and there was no relation between gas production and stool consistency, although stools were more likely to be malodorous when concentrations of H2S and/or CH3SH were high. CONCLUSIONS: Gas release by infant feces is strongly influenced by an infant's diet. Of particular interest are differences in production of the highly toxic sulfur gases, H2S and CH3SH, because of the role that these gases may play in certain intestinal disorders of infants. PMID- 11429514 TI - Relation between serum insulinlike growth factor-1, insulinlike growth factor binding protein-2, and insulinlike growth factor binding protein-3 and nutritional intake in premature infants with bronchopulmonary dysplasia. AB - BACKGROUND: The usefulness of serum insulinlike growth factor (IGF)-system peptide measurement to assess the adequacy of nutritional intake in premature infants with chronic lung disease bronchopulmonary dysplasia (BPD) was assessed. METHODS: Twenty-nine premature infants had serial measurements taken of their serum IGF-1, insulinlike growth factor binding protein (IGFBP)-2, and IGFBP-3 concentrations between 2 and 6 weeks of age. Regression analyses were used to examine the relation between nutritional parameters and IGF-1, IGFBP-2, and IGFBP 3 concentrations in premature infants with and without BPD. RESULTS: The group of infants with BPD (n = 12) did not differ from infants without BPD (n = 17) in gestational age or weight at entry, but gained less weight during the study period. In infants without BPD, IGF-1 correlated positively with protein intake (r = 0.50) and caloric intake (r = 0.41) over the 3 days before sample collection and with weight change over the previous week (r = 0.46). In contrast, infants with BPD showed a significant correlation between IGF-1 and weight change (r = 0.54) only. There was a significant negative correlation between IGFBP-2 and protein intake in infants without BPD (r = -0.50) and in infants with BPD (r = 0.41). Negative correlations between IGFBP-2 and both weight change (r = -0.64) and caloric intake (r = -0.43) over the previous week were found only in the group of infants without BPD. IGFBP-3 correlated positively with weight changes and protein intake in both groups but correlated with caloric intake only in the group without BPD. Multiple regression analyses were used to determine significant independent variables associated with IGF-1, IGFBP-2, and IGFBP-3. In infants without BPD, significant independent predictors of IGFBP-2 were 7-day weight change and 2-day protein intake; 3-day caloric intake was the only significant independent predictor for IGFBP-3. For infants with BPD, 3-day weight gain was the only independent variable associated with serum IGF-1. Protein intake in the week before sample collection was an independent predictor of IGFBP 2 and 3-day weight change and 2-day protein intake were independent predictors of IGFBP-3. CONCLUSIONS: These results confirm that changes in serum IGF-1, IGFBP-2, and IGFBP-3 reflect the nutritional status of premature infants and demonstrate that the relation between these proteins and nutritional intake differs in premature infants with and without BPD. Refinement of these observations by future studies may permit a more accurate determination of the protein and caloric intake sufficient for growth and repair after injury in premature infants with lung disease. PMID- 11429515 TI - Intestinal and systemic infection, HIV, and mortality in Zambian children with persistent diarrhea and malnutrition. AB - BACKGROUND: Persistent diarrhea-malnutrition syndrome is a complex of infection and immune failure that involves protein, calorie and micronutrient depletion, and metabolic disturbances. We report an analysis of the impact of HIV infection on infectious disease, clinical presentation, and mortality in Zambian children with persistent diarrhea and malnutrition. METHODS: Two hundred children (94 boys and 106 girls, 6-24 months old) were examined on admission to the malnutrition ward of University Teaching Hospital in Lusaka, Zambia. There was then 1 month of follow-up. RESULTS: Antibodies to HIV were found in 108 of the children (54%). The common intestinal infections (Cryptosporidium parvum [26%] and nontyphoid Salmonella spp [18%]), septicemia (17%), and pulmonary tuberculosis confirmed by gastric lavage (13.5%) were not significantly more common in HIV-seropositive than in HIV-seronegative children. HIV-seropositive children were more likely to have marasmus whereas HIV-seronegative children were more likely to have kwashiorkor. Weight-for-age z scores at nadir (postedema) were lower in HIV seropositive children (median, -4.4; interquartile range [IQR], -5.0 to -3.8) than in HIV-seronegative children (median, -3.7; IQR, -4.2 to -3.1; P < 0.0001). Height-for-age and weight-for-height z scores and mid-upper arm circumference showed a similar difference. Of the 200 children, 39 (19.5%) died within 28 days; cryptosporidiosis and marasmus were the only independent predictors of death. CONCLUSIONS: Although intestinal and systemic infections did not differ for HIV seropositive and HIV-seronegative children, HIV influenced nutritional states of all children. Cryptosporidiosis and marasmus were associated with higher mortality. PMID- 11429516 TI - Nutritional efficacy of preterm formula with a partially hydrolyzed protein source: a randomized pilot study. AB - BACKGROUND: Decreased nitrogen levels, calcium intestinal absorption rates, and plasma amino acid imbalances were reported for preterm infants who were fed partially hydrolyzed preterm formulas. In this pilot study, we evaluated a new formula with modified nitrogen and calcium sources. METHODS: During their second week of life, 16 preterm infants were randomly assigned to one of two groups: 9 were fed the new partially hydrolyzed formula and 7 were fed a conventional formula. Nutrient balance was performed at the end of the first month of life. Amino acid concentrations and anthropometric parameters were measured at theoretical term. RESULTS: Birth weight and gestational age (mean +/- SD) were similar in the two groups (28.9 +/- 7.0 weeks and 1183 +/- 242 g vs. 27.7 +/- 1.0 weeks and 1139 +/- 162 g). Median nitrogen absorption rates (85% vs. 89%; P = 0.03) and biological values (59% vs. 69%; P = 0.13) were lower for infants who were fed the new formula than for those fed the conventional formula. After correction for difference in nitrogen intake, there was no significant difference in nitrogen retained between the two groups (P = 0.11). Plasma amino acid concentrations were also similar in the two groups. Median calcium absorption tended to be higher in the new-formula group than in the conventional-formula group (54% vs. 45%, P = 0.19). At theoretical term, infants fed the conventional formula were heavier than infants fed the new formula (3559 +/- 362 g vs. 3193 +/ 384 g, P = 0.04). CONCLUSIONS: Because nitrogen content is 10% higher in hydrolyzed-protein formula than in entire-protein formula, appropriate nitrogen retention, plasma amino acid profile, and mineral use can be achieved with the new partially hydrolyzed formula. Further studies with larger groups are needed to evaluate the effect on growth. PMID- 11429517 TI - An epidemiological survey of hepatitis C virus infection in Italian children in the decade 1990-1999. AB - BACKGROUND: A retrospective-prospective survey of Italian children with hepatitis C virus (HCV) infection was planned in 1998 to explore the epidemiologic features of infection during the past decade. METHODS: Anti-HCV-positive patients (or HCV RNA-positive infants) aged 1 month to 16 years, consecutively observed in 20 pediatric Institutions, were considered. An anonymous epidemiologic questionnaire based on clinical records was used. RESULTS: From 1990 through March 1999, 606 patients were observed (296 boys, average age 5.8 years). Maternal infection (46% of cases) and blood transfusions (34%) were the most frequent risk factors. Of 279 infected mothers, 61% did not recall a putative source of infection (by history, many could possibly have had exposure through routes such as therapeutic injections with nondisposable material), whereas 94 (34%) admitted drug abuse, including 49 (17%) coinfected with human immunodeficiency virus (HIV). Only 157 (26%) children were born after 1991: 90% of their mothers were infected (11% were HIV coinfected vs. 25% mothers of older children, P < 0.01). CONCLUSIONS: Maternal infection is a prominent source of pediatric HCV infection in Italy. The fact that most mothers had a history of covert exposure to HCV, probably through percutaneous routes that are no longer operating, and that the number of those with HIV coinfection has decreased suggests that the frequency of pediatric infection could decrease in the future. PMID- 11429518 TI - Extent of intestinal damage in the developing chick embryo after repetitive hypoxia under normoxic or hyperoxic conditions. AB - BACKGROUND: Episodes of hypoxia and reperfusion play an important role in the development of intestinal damage during perinatal development. The aim of this study was to investigate the histopathology of the intestine in the developing chick embryo after exposure to repetitive hypoxia and recovery under two different conditions: normoxic and hyperoxic (60% O2). METHODS: Chick embryos were exposed to 5 minutes of hypoxia. This was repeated six times with a recovery period of 15 minutes under normoxic conditions (21% O2) for chick embryos in test group 1 (TG1) and under hyperoxic conditions (100% O2) for chick embryos in test group 2 (TG2), from day 11 until day 20. Chick embryos that recovered under hyperoxic conditions (100% O2) were previously incubated under hyperoxic conditions (60% O2 for 24 hours). Histologic evaluation of the ileum was performed at different times after the interventions (2, 4, 8, 16, and 24 hours). RESULTS: In both test groups, only chick embryos older than 19 days showed intestinal damage. Intestinal histology on day 19 showed vasodilation of villus capillaries (10% in TG1 and 15% in TG2), necrosis in the top of the villi (29% in TG1 and 30% in TG2), and necrosis with preservation of base of the crypts (2% in TG1) and transmucosal necrosis (2% in TG2). CONCLUSIONS: Significant histologic changes, compared with the control group, were only found in chick embryos that were studied 2 hours after the interventions. Furthermore, recovery under hyperoxic conditions did not cause more intestinal damage compared with recovery under normoxic conditions. PMID- 11429519 TI - Quality of life in inflammatory bowel disease: a cross-cultural comparison of English and Canadian children. AB - BACKGROUND: Any disease and its treatment has an important impact on health related quality of life for affected individuals. There have been few previous studies on the quality of life for children with inflammatory bowel disease (IBD). METHODS: A cross-cultural comparison was performed to determine whether the concerns of children with IBD in the United Kingdom are ranked similarly to those of children with IBD in Canada. An item reduction questionnaire, developed from interviews with Canadian children with IBD, was scored by 53 British children with IBD for importance and frequency, as a questionnaire had been scored previously by 117 Canadian children. RESULTS: There was a significant correlation between the mean scores (r = 0.831, P < 0.001) and ranks (r = 0.801, P < 0.001) for the 96 questions, and 43 of the 50 highest-ranking concerns corresponded for both populations. Confidence interval analysis showed a significant difference between the mean values for 21 of the 96 items; 20 of these 21 were ranked higher in the United Kingdom than they had been in Canada, suggesting that the frequency and/or degree of concern was greater for the British children with IBD. CONCLUSIONS: Health-related concerns of British children with Crohn disease and ulcerative colitis correlate closely with those of Canadian children with those diseases. Further studies are needed to determine the sensitivity of individual questions, the most appropriate wording of these questions, and the optimal length for a proposed instrument to assess quality of life in children with IBD. PMID- 11429520 TI - 13C mixed triglyceride breath test: a noninvasive method to assess lipase activity in children. AB - BACKGROUND: Results from the 13C mixed triglyceride (MTG) breath test correlate with duodenal lipase activity in adults. This noninvasive test is a potential screening and diagnostic tool for children with fat malabsorption. The aim of this study was to adapt the methodology of the MTG breath test to study test meals and sampling methods and to define normal values for healthy children of all age groups; premature and full-term infants have similar pancreatic lipase deficiencies. METHODS: After parental consent was obtained, 12 premature infants (< 37 weeks gestation and with body weights > 2 kg), 12 full-term infants (1-6 months old), 20 children (3-10 years old), and 20 teenagers (11-17 years old) were tested. All children were thriving well, had no gastrointestinal or respiratory problems, and had not received any medication that contained natural 13C. For the premature and full-term infants, a formula was prepared that had a low and stable natural 13C content mixed with 100 mg 13C-labeled MTG (1,3 distearyl, 2-[13C-carboxyl] octanoyl glycerol) and 1 g polyethylene-glycol 3350. The best accepted test meal for children over 3 years old was a slice of white bread with 5 g butter and 15 g chocolate paste, mixed with 250 mg 13C-labeled MTG, and a glass of 100 mL whole-fat milk. Children over 3 years old were able to blow through a straw in a vacutainer for collecting the breath samples. In children under 3 years old, expired air was collected by aspirating breath via a nasal prong. Carbon dioxide production was calculated according to weight, age, and sex. RESULTS: For healthy pediatric control participants, the mean values for cumulative excretion of 13CO2 as a percentage of the administered dose after 6 hours were 23.9 +/- 5.2% in premature infants, 31.9 +/- 7.7% in full-term infants, 32.5 +/- 5.3% in children, and 28.0 +/- 5.4% in teenagers. The mean value for healthy adults is 35.6% with a lower reference limit of 22.8%. CONCLUSIONS: Age-specific test meals and breath-sampling techniques for the MTG breath test were defined. The mean values for different age groups may serve as guidelines for normal values in the pediatric population. The cumulative values for expired 13CO2 were above the lower limit for adults, which suggests that preduodenal lipases compensates for pancreatic lipase deficiency in premature and full-term infants. PMID- 11429521 TI - Extremely short small bowel induces focal tubulointerstitial fibrosis. AB - BACKGROUND: Arginine becomes an essential amino acid after massive resection of the small bowel as a result of decreased biosynthesis of citrulline in the remaining small bowel. It is also reported that nitric oxide (NO) is synthesized from l-arginine by NO synthase (NOS), and NO is involved in the regulation of blood flow in the kidney. The authors observed a patient with an extremely short small bowel, showing focal tubulointerstitial fibrosis. The experiment was designed to clarify whether massive small bowel resection (SBR) produces focal tubulointerstitial fibrosis in the kidney. METHODS: An experimental study was performed using 4-week-old rats with 90% proximal SBR either with or without arginine supplementation for 6 weeks after surgery. RESULTS: In rats without arginine supplementation, low plasma levels of citrulline and arginine increased urinary excretion of orotate, and focal tubulointerstitial fibrosis was observed 6 weeks after 90% SBR. The data from plasma amino acid chromatography and increased excretion of urinary orotate suggested the presence of arginine deficiency. The kidney pathology was similar to that of our patient. Rats with arginine supplementation after 90% SBR and pair-fed control rats without 90% SBR showed almost normal glomeruli and tubulointerstitium. CONCLUSIONS: Experimental study strongly suggests that arginine deficiency causes focal tubulointerstitial fibrosis in the kidney after massive SBR. PMID- 11429522 TI - Lipoprotein lipase deficiency: benefits and limitations of a novel therapeutic surgical approach. PMID- 11429523 TI - Biliary microlithiasis: a cause of idiopathic pancreatitis in childhood. PMID- 11429524 TI - Primary non-Hodgkin lymphoma of the extrahepatic biliary tract and gallbladder in a child. PMID- 11429525 TI - Coexistence of pancreas divisum and septate common channel in a child with choledochal cyst. PMID- 11429526 TI - Fulminant hepatitis associated with Schistosoma mansoni. PMID- 11429527 TI - Primary gastric lymphoma associated with Helicobacter pylori in a child. PMID- 11429528 TI - Infectious strictures requiring esophageal replacement in children. PMID- 11429529 TI - Lansoprazole triple therapy for Turkish children with Helicobacter pylori infections. PMID- 11429530 TI - Dual-energy x-ray absorptiometry: a new insight into geophagy. PMID- 11429531 TI - Foot and mouth. PMID- 11429532 TI - Peer-to-peer: reply to the Editor. PMID- 11429533 TI - Vascular targeting and antigen presentation. PMID- 11429534 TI - The lymphocyte in immunology: from James B. Murphy to James L. Gowans. AB - Between 1912 and 1921, James Murphy established conclusively the role of the lymphocyte in tissue and tumor graft rejection and in protection against infection. Contemporary mainstream immunology paid little attention to these findings, until the lymphocyte was "rediscovered" with the advent of modern cellular immunology after the mid-1950s. PMID- 11429535 TI - ICOS costimulation: It's not just for TH2 cells anymore. AB - A current paradigm has ICOS participating in TH2 costimulation. New data indicates ICOS regulates not only TH2 cells, but also TH1s. PMID- 11429536 TI - Kissing cousins: immunological and neurological synapses. AB - Immunological synapse formation is essential for T cell activation. A recent paper in Science reports that immunological and neurological synapses utilize a common molecule, agrin. PMID- 11429537 TI - To B or not to B: TNF family signaling in lymphocytes. AB - BLyS and family are known to affect B cells in a positive fashion. Knock-outs of BLyS receptors indicate some new functions, including negative regulation by one BLyS receptor, TACI. PMID- 11429538 TI - Unraveling the mysteries of gammadelta T cell recognition. AB - gammadelta T cells are the misunderstood siblings of the antigen receptor family. A recent paper in Nature that describes the crystal structure of a gammadelta TCR should initiate a clearer understanding of these enigmatic cells. PMID- 11429539 TI - B cells with the guts to switch. AB - Expression of mIgM was thought to be esential for the differentiation of B cells expressing antibodies of other classes. New evidence suggests isotype class switching to IgA can occur in the absence of mIgM. PMID- 11429541 TI - Dendritic cell lineage, plasticity and cross-regulation. AB - Dendritic cells (DCs) are professional antigen-presenting cells that have an extraordinary capacity to stimulate naive T cells and initiate primary immune responses. Here we review progress in understanding the additional functions of DCs in regulating the types of T cell-mediated immune responses and innate immunity to microbes. In addition, evidence for the existence of myeloid and lymphoid DC lineages and their different functions are summarized. We propose that the diverse functions of DCs in immune regulation are dictated by the instructions they received during innate immune responses to different pathogens and from their evolutionary lineage heritage. PMID- 11429542 TI - Importance of ICOS-B7RP-1 costimulation in acute and chronic allograft rejection. AB - Primary T cell activation requires B7-CD28 and CD40-CD154 costimulation, but effector T cell functions are considered to be largely independent of these costimulatory pathways. Although blockade of costimulation with cytolytic T lymphocyte-associated antigen 4-immunoglobulin (CTLA-4-Ig) or monoclonal antibody (mAb) to CD154 prolongs allograft survival, chronic rejection follows, which suggests that additional key costimulatory pathways are active in vivo. We found that both antibody to inducible costimulator (anti-ICOS) and an ICOS-Ig fusion protein suppressed intragraft T cell activation and cytokine expression and prolonged allograft survival in a manner similar to that in ICOS-/- allograft recipients. The combination of anti-ICOS therapy and cyclosporin A led to permanent engraftment. In addition, ICOS-B7RP-1 costimulation was required for the development of chronic rejection after CD40-CD154 blockade. These data demonstrate a key role for the ICOS-B7RP-1 pathway in acute and chronic rejection and highlight the benefits of targeting this pathway in combination with the use of conventional immunosuppressive agent. PMID- 11429543 TI - ICOS is critical for T helper cell-mediated lung mucosal inflammatory responses. AB - We examined the requirement for and cooperation between CD28 and inducible costimulator (ICOS) in effective T helper (TH) cell responses in vivo. We found that both CD28 and ICOS were critical in determining the outcome of an immune response; cytolytic T lymphocyte-associated antigen 4-immunoglobulin (CTLA-4-Ig), ICOS-Ig and/or a neutralizing ICOS monoclonal antibody attenuated T cell expansion, TH2 cytokine production and eosinophilic inflammation. CD28-dependent signaling was essential during priming, whereas ICOS-B7RP-1 regulated TH effector responses, and the up-regulation of chemokine receptors that determine T cell migration. Our data suggests a scenario whereby both molecules regulate the outcome of the immune response but play separate key roles: CD28 primes T cells and ICOS regulates effector responses. PMID- 11429544 TI - The costimulatory molecule ICOS plays an important role in the immunopathogenesis of EAE. AB - The inducible costimulatory molecule (ICOS) is expressed on activated T cells and participates in a variety of important immunoregulatory functions. After the induction of experimental allergic encephalomyelitis in SJL mice with proteolipid protein (PLP), brain ICOS mRNA and protein were up-regulated on infiltrating CD3+ T cells before disease onset. ICOS blockade during the efferent immune response (9-20 days after immunization) abrogated disease, but blockade during antigen priming (1-10 days after immunization) exacerbated disease. Upon culture with PLP and compared with immunized controls, splenocytes produced either decreased interferon-gamma (IFN-gamma, in efferent blockade) or excessive IFN-gamma (in priming blockade). PLP-specific immunoglobulin G1 was decreased in animals treated with anti-ICOS during antigen priming, but not in other groups. PMID- 11429545 TI - Lipid mediator class switching during acute inflammation: signals in resolution. AB - Leukotrienes (LTs) and prostaglandins (PGs) amplify acute inflammation, whereas lipoxins (LXs) have unique anti-inflammatory actions. Temporal analyses of these eicosanoids in clinical and experimental exudates showed early coordinate appearance of LT and PG with polymorphonuclear neutrophil (PMN) recruitment. This was followed by LX biosynthesis, which was concurrent with spontaneous resolution. Human peripheral blood PMNs exposed to PGE2 (as in exudates) switched eicosanoid biosynthesis from predominantly LTB4 and 5-lipoxygenase (5-LO) initiated pathways to LXA4, a 15-LO product that "stopped" PMN infiltration. These results indicate that first-phase eicosanoids promote a shift to anti inflammatory lipids: functionally distinct lipid-mediator profiles switch during acute exudate formation to "reprogram" the exudate PMNs to promote resolution. PMID- 11429546 TI - The essential role of MEKK3 in TNF-induced NF-kappaB activation. AB - Activation of IkappaB kinase (IKK) is the key step in stimulation of the transcription factor NF-kappaB, which regulates many genes in the inflammatory response pathway. The molecular mechanism that underlies IKK activation in response to tumor necrosis factor (TNF) is still unknown. Using mitogen-activated protein kinase kinase kinase 3 (MEKK3)-deficient fibroblast cells, we found that MEKK3 plays a critical role in TNF-induced NF-kappaB activation. We have shown that MEKK3 is required for IKK activation and functions downstream of receptor interacting protein (RIP) and TNF receptor- associated factor 2. We have also shown that MEKK3 interacts with RIP and directly phosphorylates IKK. The kinase activity of MEKK3 is pivotal to its function and, therefore, MEKK3 links RIP and IKK in TNF-induced NF-kappaB activation. PMID- 11429547 TI - IgA production without mu or delta chain expression in developing B cells. AB - Surface, membrane-bound, immunoglobulin M (IgM) or IgD expression early in B cell ontogeny is considered essential for the differentiation of antibody-producing cells in mammals; only in IgM+ B cells is the heavy chain locus rearranged to express antibodies of other classes. We show here that IgA is selectively expressed in muMT mice, which lack IgM or IgD expression and have a pro-B cell developmental block. muMT IgA binds proteins of commensal intestinal bacteria and is weakly induced by Salmonella infection, although not through conventional immunization. This muMT IgA pathway requires extrasplenic peripheral lymphoid tissues and may be an evolutionarily primitive system in which immature B cells switch to IgA production at peripheral sites. PMID- 11429548 TI - TACI-ligand interactions are required for T cell activation and collagen-induced arthritis in mice. AB - Interactions of the tumor necrosis factor superfamily members B lymphocyte stimulator (BLyS) and a proliferation-inducing ligand (APRIL) with their receptors-transmembrane activator and CAML interactor (TACI) and B cell maturation molecule (BCMA)-on B cells play an important role in the humoral immune response. Whereas BCMA is restricted to B cells, TACI is also expressed on activated T cells; we show here that TACI-Fc blocks the activation of T cells in vitro and inhibits antigen-specific T cell activation and priming in vivo. In a mouse model for rheumatoid arthritis (RA), an autoimmune disease that involves both B and T cell components, TACI-Fc treatment substantially inhibited inflammation, bone and cartilage destruction and disease development. Thus, BLyS and/or APRIL are important not only for B cell function but for T cell-mediated immune responses. Inhibition of these ligands might have therapeutic benefits for autoimmune diseases, such as RA, that involve both B and T cells. PMID- 11429549 TI - Activation and accumulation of B cells in TACI-deficient mice. AB - The tumor necrosis factor (TNF)-related ligand B lymphocyte stimulator (BLyS) binds two TNF receptor family members, transmembrane activator and calcium modulating and cyclophilin ligand interactor (TACI) and B cell maturation molecule (BCMA). Mice that are transgenic for BLyS show B cell accumulation, activation and autoimmune lupus-like nephritis. The existence of at least two distinct BLyS receptors raises the question of the relative contribution of each to B cell functions. We therefore generated mice that were deficient in TACI. TACI-/- mice showed increased B cell accumulation and marked splenomegaly. Isolated TACI-/- B cells hyperproliferated and produced increased amounts of immunoglobulins in vitro. In vivo antigen challenge resulted in enhanced antigen specific antibody production. Thus, TACI may play an unexpected inhibitory role in B cell activation that helps maintain immunological homeostasis. PMID- 11429550 TI - ER aminopeptidases generate a unique pool of peptides for MHC class I molecules. AB - We define here the specificity and significance of proteases in the endoplasmic reticulum (ER) that generate peptides for presentation by major histocompatibility complex (MHC) class I molecules. We show that aminopeptidases efficiently trimmed all residues except proline that flank the NH2-termini of antigenic precursors in the ER and caused an accumulation of X-P-Xn peptides. An aminopeptidase inhibitor blocked peptide trimming in the ER and, consequently, the generation of peptide-loaded MHC molecules. Peptide trimming in the ER is therefore a key step in the MHC class I antigen-processing pathway and also explains the paradox of why many MHC class I molecules display peptides with the X-P-Xn motif despite the inability of the transporter associated with antigen processing to transport such peptides from the cytoplasm. PMID- 11429551 TI - CIITA coordinates multiple histone acetylation modifications at the HLA-DRA promoter. AB - We present here an in vivo view of major histocompatibility complex (MHC) class II promoter assembly, nucleosome modifications and gene expression mediated by the class II transactivator (CIITA). Acetylation and deacetylation of histones H3 and H4 at the HLA-DRA promoter were found to occur during a time-course that depended on CIITA expression and binding. Expression of a CIITA mutant, which lacked the activation domain, induced H4 but not H3 histone acetylation. This suggested that multiple histone acetyltransferase activities are associated with MHC class II expression. H4 acetylation was mapped to Lys8, which implicated several histone acetyltransferases as possible modulators of this activity. PMID- 11429554 TI - Lessons from an epidemic. PMID- 11429555 TI - Defending tax-funded navigation. PMID- 11429556 TI - Stem-cell debate heats up as Bush ponders policy. PMID- 11429557 TI - Physicist nominated as science adviser for US administration. PMID- 11429558 TI - Budget proposal casts doubt over physics portal's future. PMID- 11429559 TI - Fear of bias puts spotlight on drug approval. PMID- 11429561 TI - Europe implements package deals for space station. PMID- 11429560 TI - US army advised to soldier on with biotechnology. PMID- 11429562 TI - Poor nations push for right to produce cheap medicines. PMID- 11429563 TI - Curbs at NIH frustrate travel plans. PMID- 11429564 TI - India blocks sale of transgenic cotton seeds. PMID- 11429566 TI - Brane new world. PMID- 11429567 TI - Vital signs. PMID- 11429568 TI - 'Espionage' charge may be based on a misunderstanding of rules. PMID- 11429569 TI - You can help rare plants survive in the cities. PMID- 11429570 TI - Undergraduates must learn to communicate. PMID- 11429571 TI - Following the lead of pioneering teachers. PMID- 11429572 TI - Brazil's WTO case has no bearing on AIDS. PMID- 11429578 TI - Archaeology of type. PMID- 11429579 TI - Predicting the future. PMID- 11429580 TI - Are all cancer genes equal? PMID- 11429581 TI - Planetary candidates. PMID- 11429582 TI - Repopulating the RNA world. PMID- 11429583 TI - Martian motion. PMID- 11429584 TI - Glowing switches. PMID- 11429585 TI - A bottleneck for optical fibres. PMID- 11429587 TI - T cells and tumours. PMID- 11429588 TI - Catch the atom. PMID- 11429590 TI - Genes lost during evolution. PMID- 11429591 TI - Night-time predation by Steller sea lions. AB - New insight into the feeding habits of these mammals will help conservation attempts. PMID- 11429592 TI - Failure to correct murine muscular dystrophy. PMID- 11429593 TI - Formation of ozone by fireworks. PMID- 11429594 TI - Subcellular positioning of small molecules. PMID- 11429595 TI - Specific protection against breast cancers by cyclin D1 ablation. AB - Breast cancer is the most common malignancy among women. Most of these cancers overexpress cyclin D1, a component of the core cell-cycle machinery. We previously generated mice lacking cyclin D1 using gene targeting. Here we report that these cyclin D1-deficient mice are resistant to breast cancers induced by the neu and ras oncogenes. However, animals lacking cyclin D1 remain fully sensitive to other oncogenic pathways of the mammary epithelium, such as those driven by c-myc or Wnt-1. Our analyses revealed that, in mammary epithelial cells, the Neu-Ras pathway is connected to the cell-cycle machinery by cyclin D1, explaining the absolute dependency on cyclin D1 for malignant transformation in this tissue. Our results suggest that an anti-cyclin D1 therapy might be highly specific in treating human breast cancers with activated Neu-Ras pathways. PMID- 11429596 TI - Gravitational microlensing by low-mass objects in the globular cluster M22. AB - Gravitational microlensing offers a means of determining directly the masses of objects ranging from planets to stars, provided that the distances and motions of the lenses and sources can be determined. A globular cluster observed against the dense stellar field of the Galactic bulge presents ideal conditions for such observations because the probability of lensing is high and the distances and kinematics of the lenses and sources are well constrained. The abundance of low mass objects in a globular cluster is of particular interest, because it may be representative of the very early stages of star formation in the Universe, and therefore indicative of the amount of dark baryonic matter in such clusters. Here we report a microlensing event associated with the globular cluster M22. We determine the mass of the lens to be 0.13(+0.03)(-0.02) solar masses. We have also detected six events that are unresolved in time. If these are also microlensing events, they imply that a non-negligible fraction of the cluster mass resides in the form of free-floating planetary-mass objects. PMID- 11429597 TI - Sub-poissonian loading of single atoms in a microscopic dipole trap. AB - The ability to manipulate individual atoms, ions or photons allows controlled engineering of the quantum state of small sets of trapped particles; this is necessary to encode and process information at the quantum level. Recent achievements in this direction have used either trapped ions or trapped photons in cavity quantum-electrodynamical systems. A third possibility that has been studied theoretically is to use trapped neutral atoms. Such schemes would benefit greatly from the ability to trap and address individual atoms with high spatial resolution. Here we demonstrate a method for loading and detecting individual atoms in an optical dipole trap of submicrometre size. Because of the extremely small trapping volume, only one atom can be loaded at a time, so that the statistics of the number of atoms in the trap, N, are strongly sub-poissonian (DeltaN2 approximately 0.5N). We present a simple model for describing the observed behaviour, and we discuss the possibilities for trapping and addressing several atoms in separate traps, for applications in quantum information processing. PMID- 11429598 TI - Nonlinear limits to the information capacity of optical fibre communications. AB - The exponential growth in the rate at which information can be communicated through an optical fibre is a key element in the 'information revolution'. However, as for all exponential growth laws, physical limits must be considered. The nonlinear nature of the propagation of light in optical fibre has made these limits difficult to elucidate. Here we use a key simplification to investigate the theoretical limits to the information capacity of an optical fibre arising from these nonlinearities. The success of our approach lies in relating the nonlinear channel to a linear channel with multiplicative noise, for which we are able to obtain analytical results. In fundamental distinction to linear channels with additive noise, the capacity of a nonlinear channel does not grow indefinitely with increasing signal power, but has a maximal value. The ideas presented here may have broader implications for other nonlinear information channels, such as those involved in sensory transduction in neurobiology. These have been often examined using additive noise linear channel models but, as we show here, nonlinearities can change the picture qualitatively. PMID- 11429599 TI - Control of conformational and interpolymer effects in conjugated polymers. AB - The role of conjugated polymers in emerging electronic, sensor and display technologies is rapidly expanding. In spite of extensive investigations, the intrinsic spectroscopic properties of conjugated polymers in precise conformational and spatial arrangements have remained elusive. The difficulties of obtaining such information are endemic to polymers, which often resist assembly into single crystals or organized structures owing to entropic and polydispersity considerations. Here we show that the conformation of individual polymers and interpolymer interactions in conjugated polymers can be controlled through the use of designed surfactant poly(p-phenylene-ethynylene) Langmuir films. We show that by mechanically inducing reversible conformational changes of these Langmuir monolayers, we can obtain the precise interrelationship of the intrinsic optical properties of a conjugated polymer and a single chain's conformation and/or interpolymer interactions. This method for controlling the structure of conjugated polymers and establishing their intrinsic spectroscopic properties should permit a more comprehensive understanding of fluorescent conjugated materials. PMID- 11429600 TI - Biodegradation of oil in uplifted basins prevented by deep-burial sterilization. AB - Biodegradation of crude oil by bacterial activity--which has occurred in the majority of the Earth's oil reserves--is known to reduce greatly the quality of petroleum in reservoirs. For economically successful prospecting for oil, it is therefore important to understand the processes and conditions in geological formations that lead to oil biodegradation. Although recent studies speculate that bacterial activity can potentially occur up to temperatures as high as 150 degrees C (refs 3, 4), it is generally accepted that effective petroleum biodegradation over geological timescales generally occurs in reservoirs with temperatures below 80 degrees C (ref. 2). This appears, however, to be at odds with the observation that non-degraded oils can still be found in reservoirs below this temperature. Here we compile data regarding the extent of oil biodegradation in several oil reservoirs, and find that the extensive occurrence of non-biodegraded oil in shallow, cool basins is restricted to those that have been uplifted from deeper, hotter regions of the Earth. We suggest that these petroleum reservoirs were sterilized by heating to a temperature around 80-90 degrees C during deep burial, inactivating hydrocarbon-degrading organisms that occur in the deep biosphere. Even when such reservoirs are subsequently uplifted to much cooler regions and filled with oil, degradation does not occur, implying that the sterilized sediments are not recolonized by hydrocarbon-degrading bacteria. PMID- 11429601 TI - Mineral disequilibrium in lavas explained by convective self-mixing in open magma chambers. AB - Characteristic features of many porphyritic andesite and dacite lavas are that they are rich in crystals and display a range of disequilibrium features, including reversely zoned crystals, resorption surfaces, wide ranges of mineral compositions and minerals which are not in equilibrium with the surrounding rock matrix. These features are often interpreted as evidence of the mixing of magmas of contrasting composition, temperature and origin. Here, however, we propose that such features can also be caused by convection within a magma body with a single composition, that is heated from below and cooled from above. We describe petrological observations of andesite lava erupted at the Soufriere Hills volcano, Montserrat, which indicate a heating event and the intermingling of crystals that have very different thermal histories. We present experimental data on a representative groundmass composition of this lava, which indicate that it is difficult to explain the calcic compositions of plagioclase overgrowth rims and microphenocrysts unless parts of the magma were at temperatures much higher than the inferred average temperature. The concept of convective self-mixing allows us to explain the occurrence of compositions of minerals that apparently cannot coexist under equilibrium conditions. PMID- 11429602 TI - Anaerobic benzene oxidation coupled to nitrate reduction in pure culture by two strains of Dechloromonas. AB - Benzene contamination is a significant problem. It is used in a wide range of manufacturing processes and is a primary component of petroleum-based fuels. Benzene is a hydrocarbon that is soluble, mobile, toxic and stable, especially in ground and surface waters. It is poorly biodegraded in the absence of oxygen. However, anaerobic benzene biodegradation has been documented under various conditions. Although benzene biomineralization has been demonstrated with nitrate, Fe(III), sulphate or CO2 as alternative electron acceptors, these studies were based on sediments or microbial enrichments. Until now there were no organisms in pure culture that degraded benzene anaerobically. Here we report two Dechloromonas strains, RCB and JJ, that can completely mineralize various mono aromatic compounds including benzene to CO2 in the absence of O2 with nitrate as the electron acceptor. This is the first example, to our knowledge, of an organism of any type that can oxidize benzene anaerobically, and we demonstrate the potential applicability of these organisms to the treatment of contaminated environments. PMID- 11429603 TI - Life-history traits of voles in a fluctuating population respond to the immediate environment. AB - Life-history traits relating to growth and reproduction vary greatly among species and populations and among individuals within populations. In vole populations, body size and age at maturation may vary considerably among locations and among years within the same location. Individuals in increasing populations are typically larger and start reproduction earlier in the spring than those in declining populations. The cause of such life-history variation within populations has been subject of much discussion. Much of the controversy concerns whether the memory of past conditions, leading to delayed effects on life-history traits, resides in the environment (for example, predators, pathogens or food) or intrinsically within populations or individuals (age distribution, physiological state, genetic or maternal effects). Here we report from an extensive field transplant experiment in which voles were moved before the breeding season between sites that differed in average overwintering body mass. Transplanted voles did not retain the characteristics of their source population, and we demonstrate an over-riding role of the immediate environment in shaping life-history traits of small rodents. PMID- 11429604 TI - Protein dispensability and rate of evolution. AB - If protein evolution is due in large part to slightly deleterious amino acid substitutions, then the rate of evolution should be greater in proteins that contribute less to individual fitness. The rationale for this prediction is that relatively dispensable proteins should be subject to weaker purifying selection, and should therefore accumulate mildly deleterious substitutions more rapidly. Although this argument was presented over twenty years ago, and is fundamental to many applications of evolutionary theory, the prediction has proved difficult to confirm. In fact, a recent study showed that essential mouse genes do not evolve more slowly than non-essential ones. Thus, although a variety of factors influencing the rate of protein evolution have been supported by extensive sequence analysis, the relationship between protein dispensability and evolutionary rate has remained unconfirmed. Here we use the results from a highly parallel growth assay of single gene deletions in yeast to assess protein dispensability, which we relate to evolutionary rate estimates that are based on comparisons of sequences drawn from twenty-one fully annotated genomes. Our analysis reveals a highly significant relationship between protein dispensability and evolutionary rate, and explains why this relationship is not detectable by categorical comparison of essential versus non-essential proteins. The relationship is highly conserved, so that protein dispensability in yeast is also predictive of evolutionary rate in a nematode worm. PMID- 11429605 TI - The contribution of sensory experience to the maturation of orientation selectivity in ferret visual cortex. AB - Sensory experience begins when neural circuits in the cerebral cortex are still immature; however, the contribution of experience to cortical maturation remains unclear. In the visual cortex, the selectivity of neurons for oriented stimuli at the time of eye opening is poor and increases dramatically after the onset of visual experience. Here we investigate whether visual experience has a significant role in the maturation of orientation selectivity and underlying cortical circuits using two forms of deprivation: dark rearing, which completely eliminates experience, and binocular lid suture, which alters the pattern of sensory driven activity. Orientation maps were present in dark-reared ferrets, but fully mature levels of tuning were never attained. In contrast, only rudimentary levels of orientation selectivity were observed in lid-sutured ferrets. Despite these differences, horizontal connections in both groups were less extensive and less clustered than normal, suggesting that long-range cortical processing is not essential for the expression of orientation selectivity, but may be needed for the full maturation of tuning. Thus, experience is beneficial or highly detrimental to cortical maturation, depending on the pattern of sensory driven activity. PMID- 11429606 TI - A defined range of guard cell calcium oscillation parameters encodes stomatal movements. AB - Oscillations in cytosolic calcium concentration ([Ca2+]cyt) are central regulators of signal transduction cascades, although the roles of individual [Ca2+]cyt oscillation parameters in regulating downstream physiological responses remain largely unknown. In plants, guard cells integrate environmental and endogenous signals to regulate the aperture of stomatal pores and [Ca2+]cyt oscillations are a fundamental component of stomatal closure. Here we systematically vary [Ca2+]cyt oscillation parameters in Arabidopsis guard cells using a 'calcium clamp' and show that [Ca2+]cyt controls stomatal closure by two mechanisms. Short-term 'calcium-reactive' closure occurred rapidly when [Ca2+]cyt was elevated, whereas the degree of long-term steady-state closure was 'calcium programmed' by [Ca2+]cyt oscillations within a defined range of frequency, transient number, duration and amplitude. Furthermore, in guard cells of the gca2 mutant, [Ca2+]cyt oscillations induced by abscisic acid and extracellular calcium had increased frequencies and reduced transient duration, and steady-state stomatal closure was abolished. Experimentally imposing [Ca2+]cyt oscillations with parameters that elicited closure in the wild type restored long-term closure in gca2 stomata. These data show that a defined window of guard cell [Ca2+]cyt oscillation parameters programs changes in steady-state stomatal aperture. PMID- 11429607 TI - Roles of tumour localization, second signals and cross priming in cytotoxic T cell induction. AB - The vertebrate immune system has evolved to protect against infections that threaten survival before reproduction. Clinically manifest tumours mostly arise after the reproductive years and somatic mutations allow even otherwise antigenic tumours to evade the attention of the immune system. Moreover, the lack of immunological co-stimulatory molecules on solid tumours could result in T-cell tolerance; that is, the failure of T cells to respond. However, this may not generally apply. Here we report several important findings regarding the immune response to tumours, on the basis of studies of several tumour types. First, tumour-specific induction of protective cytotoxic T cells (CTLs) depends on sufficient tumour cells reaching secondary lymphatic organs early and for a long enough duration. Second, diffusely invading systemic tumours delete CTLs. Third, tumours that stay strictly outside secondary lymphatic organs, or that are within these organs but separated from T cells by barriers, are ignored by T cells but do not delete them. Fourth, co-stimulatory molecules on tumour cells do not influence CTL priming but enhance primed CTL responses in peripheral solid tumours. Last, cross priming of CTLs by tumour antigens, mediated by major histocompatibility complex (MHC) class I molecules of antigen-presenting host cells, is inefficient and not protective. These rules of T-cell induction and maintenance not only change previous views but also rationales for anti-tumour immunotherapy. PMID- 11429608 TI - Spatio-temporal images of growth-factor-induced activation of Ras and Rap1. AB - G proteins of the Ras family function as molecular switches in many signalling cascades; however, little is known about where they become activated in living cells. Here we use FRET (fluorescent resonance energy transfer)-based sensors to report on the spatio-temporal images of growth-factor-induced activation of Ras and Rap1. Epidermal growth factor activated Ras at the peripheral plasma membrane and Rap1 at the intracellular perinuclear region of COS-1 cells. In PC12 cells, nerve growth factor-induced activation of Ras was initiated at the plasma membrane and transmitted to the whole cell body. After three hours, high Ras activity was observed at the extending neurites. By using the FRAP (fluorescence recovery after photobleaching) technique, we found that Ras at the neurites turned over rapidly; therefore, the sustained Ras activity at neurites was due to high GTP/GDP exchange rate and/or low GTPase activity, but not to the retention of the active Ras. These observations may resolve long-standing questions as to how Ras and Rap1 induce different cellular responses and how the signals for differentiation and survival are distinguished by neuronal cells. PMID- 11429609 TI - Cyclin-dependent kinases prevent DNA re-replication through multiple mechanisms. AB - The stable propagation of genetic information requires that the entire genome of an organism be faithfully replicated once and only once each cell cycle. In eukaryotes, this replication is initiated at hundreds to thousands of replication origins distributed over the genome, each of which must be prohibited from re initiating DNA replication within every cell cycle. How cells prevent re initiation has been a long-standing question in cell biology. In several eukaryotes, cyclin-dependent kinases (CDKs) have been implicated in promoting the block to re-initiation, but exactly how they perform this function is unclear. Here we show that B-type CDKs in Saccharomyces cerevisiae prevent re-initiation through multiple overlapping mechanisms, including phosphorylation of the origin recognition complex (ORC), downregulation of Cdc6 activity, and nuclear exclusion of the Mcm2-7 complex. Only when all three inhibitory pathways are disrupted do origins re-initiate DNA replication in G2/M cells. These studies show that each of these three independent mechanisms of regulation is functionally important. PMID- 11429610 TI - Multiple pathways cooperate in the suppression of genome instability in Saccharomyces cerevisiae. AB - Gross chromosome rearrangements (GCRs), such as translocations, deletion of a chromosome arm, interstitial deletions and inversions, are often observed in cancer cells. Spontaneous GCRs are rare in Saccharomyces cerevisiae; however, the existence of mutator mutants with increased genome instability suggests that GCRs are actively suppressed. Here we show by genetic analysis that these genome rearrangements probably result from DNA replication errors and are suppressed by at least three interacting pathways or groups of proteins: S-phase checkpoint functions, recombination proteins and proteins that prevent de novo addition of telomeres at double-strand breaks (DSBs). Mutations that inactivate these pathways cause high rates of GCRs and show synergistic interactions, indicating that the pathways that suppress GCRs all compete for the same DNA substrates. PMID- 11429611 TI - Crystal structure of a complex of a type IA DNA topoisomerase with a single stranded DNA molecule. AB - A variety of cellular processes, including DNA replication, transcription, and chromosome condensation, require enzymes that can regulate the ensuing topological changes occurring in DNA. Such enzymes-DNA topoisomerases-alter DNA topology by catalysing the cleavage of single-stranded DNA (ssDNA) or double stranded DNA (dsDNA), the passage of DNA through the resulting break, and the rejoining of the broken phosphodiester backbone. DNA topoisomerase III from Escherichia coli belongs to the type IA family of DNA topoisomerases, which transiently cleave ssDNA via formation of a covalent 5' phosphotyrosine intermediate. Here we report the crystal structure, at 2.05 A resolution, of an inactive mutant of E. coli DNA topoisomerase III in a non-covalent complex with an 8-base ssDNA molecule. The enzyme undergoes a conformational change that allows the oligonucleotide to bind within a groove leading to the active site. We note that the ssDNA molecule adopts a conformation like that of B-DNA while bound to the enzyme. The position of the DNA within the realigned active site provides insight into the role of several highly conserved residues during catalysis. These findings confirm various aspects of the type IA topoisomerase mechanism while suggesting functional implications for other topoisomerases and proteins that perform DNA rearrangements. PMID- 11429613 TI - Nitric oxide and the autonomic regulation of cardiac excitability. The G.L. Brown Prize Lecture. AB - Cardiac sympathetic imbalance and arrhythmia; Nitric oxide-cGMP pathway and the cholinergic modulation of cardiac excitability; Nitric oxide-cGMP pathway and the sympathetic modulation of cardiac excitability; Functional significance of nitric oxide in the autonomic regulation of cardiac excitability; Summary; References. Experimental Physiology (2001) 86.1, 1-12. PMID- 11429614 TI - Kinetics of thyroxine (T(4)) and triiodothyronine (T(3)) transport in the isolated rat heart. AB - The dynamics and kinetics of thyroid hormone transport in the isolated rat heart were examined using the modified unidirectional paired tracer dilution method. The uptake of (125)I-thyroxine ((125)I-T(4)) and (125)I-triiodothyronine ((125)I T(3)) from the extracellular space into heart cells was measured relative to the extracellular space marker (3)H-mannitol. The thyroid hormone maximal uptake was 54.4 % for (125)I-T(4) and 52.15 % for (125)I-T(3). The thyroid hormone net uptake was 25.69 % for (125)I-T(4) and 25.49 % for (125)I-T(3). Backflux from the intracellular space was 53.17 % for (125)I-T(4) and 61.59 % for (125)I-T(3). In the presence of unlabelled thyroid hormones, (125)I-T(4) and (125)I-T(3) maximal uptakes were reduced from 10.1 to 59.74 % and from 34.6 to 65.3 %, respectively, depending on the concentration of the unlabelled hormone, suggesting a saturable mechanism of the thyroid hormone uptake by the heart cells, with K(m(T4))= 105.46 microM and the maximal rate of (125)I-thyroid hormone flux from the extracellular space to heart cells (V(max(T4))) = 177.84 nM min(-1) for (125)I-T(4) uptake, and K(m(T3)) = 80.0 microM and V(max(T3)) = 118.5 nM min(-1) for (125)I-T(3) uptake. Experimental Physiology (2001) 86.1, 13-18. PMID- 11429615 TI - Raised extracellular potassium attenuates the sympathetic modulation of sino atrial node pacemaking in the isolated guinea-pig atria. AB - Intense exercise or myocardial ischaemia can significantly increase both the concentration of extracellular potassium ([K(+)](o)) and cardiac sympathetic nerve activity. Since changes in [K(+)](o) modulate membrane currents involved in sino-atrial node pacemaking, in particular the voltage-sensitive hyperpolarization-activated current (I(f)), we investigated whether raised [K(+)](o) (from 4 mM to 8 or 12 mM) could directly affect the heart rate response to cardiac sympathetic nerve stimulation (SNS). In the isolated guinea-pig atrial right stellate ganglion preparation, raised [K(+)](o) significantly decreased the maximum diastolic potential, amplitude and maximum rate of rise of the upstroke of sino-atrial node pacemaker action potentials in 8 and 12 mM [K(+)](o) (P < 0.05). At 12 mM [K(+)](o) these effects were associated with significant decreases in baseline heart rate (4 mM [K(+)](o) = 187 +/- 5 beats min(-1) (bpm); 12 mM = 144 +/- 11 bpm; P < 0.05) and the heart rate response to SNS (1, 3 and 5 Hz; P < 0.05). A 10 % increase in the baseline heart rate with sympathetic activation (3 Hz) was associated with a significant enhancement of the slope of the pacemaker diastolic depolarization at 4 mM [K(+)](o) (increased by 16 +/- 6 %; n = 7; P < 0.05), but not with raised [K(+)](o). When the I(f) current was blocked with 2 mM caesium (n = 8), 12 mM [K(+)](o) had no effect on baseline heart rate and the heart rate response to 3 Hz SNS. The heart rate response to bath-applied noradrenaline (0.01-100 microM) was significantly attenuated by 12 mM [K(+)](o) (at 4 mM [K(+)](o,) EC(50) = -6.31 +/- 0.18; at 12 mM [K(+)](o,) EC(50) = -5.80 +/- 0.10; n = 6, ANOVA, P < 0.05). In conclusion, extreme physiological levels of [K(+)](o) attenuate the positive chronotropic response to cardiac sympathetic activation due to decreased activation of the I(f) current. This is consistent with raised [K(+)](o) protecting the myocardium from potentially adverse effects of excessive noradrenaline. Experimental Physiology (2001) 86.1, 19-25. PMID- 11429616 TI - The effect of acidosis on the ECG of the rat heart. AB - We have investigated the effect of acidosis on the ECG in isolated rat heart to determine whether acidosis has marked effects on the ECG, and have used pharmacological agents to investigate possible mechanisms whereby acidosis alters the ECG. Acidosis produced a marked decrease in heart rate and an increase in P-R interval with little apparent effect on the duration of the QRS complex. The effects of acidosis did not appear to be due to acidosis-induced changes in transmitter release from severed autonomic nerve terminals within the heart. Experimental Physiology (2001) 86.1, 27-31. PMID- 11429617 TI - Interstitial fluid pressure surrounding rat mesenteric venules during changes in fluid filtration. AB - The interstitial fluid pressure (P(isf)) has been measured in the exposed superfused mesenteries of anaesthetised rats using the micropipette servo-null technique. When mesenteries were superfused with Ringer-Locke solutions, P(isf) was close to atmospheric pressure with mean +/- S.E.M. values of -0.46 +/- 0.14 cmH(2)O (n = 22). Superfusing with paraffin oil did not alter P(isf) significantly, but P(isf) could be lowered considerably by removing fluid from the upper surface of the mesentery. Measurements of P(isf) were also made in the tissues immediately outside mesenteric venules as the pressure inside these vessels and the filtration of fluid through their walls was varied. No significant changes in perivascular P(isf) could be detected even though the intravascular pressure varied from 20 to 70 cmH(2)O. Addition of histamine or the mast cell degranulating agent compound 48/80 to the superfusate had no significant effect on P(isf). The findings are relevant to experiments on the permeability of single perfused mesenteric microvessels. They strengthen the assumption, which is made in these studies, that P(isf) is close to atmospheric pressure and does not change significantly with changes in the filtration and reabsorption of fluid through the vessel walls. Experimental Physiology (2001) 86.1, 33-38. PMID- 11429618 TI - Activation of IP(3)-protein kinase C-alpha signal transduction pathway precedes the changes of plasma cholesterol, hepatic lipid metabolism and induction of low density lipoprotein receptor expression in 17-beta-oestradiol-treated rats. AB - The intracellular concentration of cholesterol is regulated by the balance between endogenous synthesis and exogenous uptake. Oestrogens have been reported to be involved in the physiological regulation of cellular cholesterol content. Relevant reports have focused on long-term responses and there is a lack of information about the relationship between the timing of the oestrogen effect and the regulation of cholesterol homeostasis. The aim of this work has been to set up a systematic picture of the short-term effects induced by oestrogen on hepatic lipid metabolism in vivo and the involvement of some relevant signal transduction pathways. At intervals after oestrogen administration (30 min to 6 h), oestrogen receptor expression and changes in liver cAMP, IP(3) and protein kinase C-alpha (PKC-alpha) were followed. Changes in the expression of the low density lipoprotein receptor at mRNA and protein levels, and of hydroxy-methyl-glutaryl CoA reductase activity have been verified. At the same time, the content of hepatic cholesterol, ubiquinone and dolichol and of plasma cholesterol have been determined. Changes of rab 5 and rab 8, small GTP-binding prenylated proteins involved in the transfer of neosynthesised proteins through the cell, have been also checked. In vivo treatment with oestradiol produced no change in cyclic AMP but a rapid increase in IP(3), increased PKC-alpha localisation on the membranes and enhanced expression of the low density lipoprotein receptor in the liver occurred. PKC inhibition completely prevented any increase in low density lipoprotein receptor mRNA in isolated and perfused rat liver. Early changes of ubiquinone and dolichol content and a later reduction in hepatic hydroxy-methyl glutaryl-CoA reductase activity and plasma cholesterol content were also detectable. A functional role of the IP(3) -protein kinase C-alpha pathway in the induction of the low density lipoprotein receptor is suggested. Experimental Physiology (2001) 86.1, 39-45. PMID- 11429619 TI - Role of nitric oxide and renal nerves in the renal responses to acute volume expansion in anaesthetized rats. AB - An investigation was undertaken into the potential role of nitric oxide (NO) and its interaction with renal sympathetic nerves in mediating renal responses to acute saline volume expansion (VE). Groups of anaesthetized Wistar rats with innervated and denervated kidneys were subjected to VE, 0.25 % body wt min-1 for 40 min, in the presence and absence of nitric oxide synthase (NOS) inhibitors, NG nitro-L-arginine-methyl-ester (L-NAME, non-selective), aminoguanidine (AG, relatively selective for inducible NOS (iNOS)), and 7-nitroindazole (7-NI, relatively selective for neuronal NOS (nNOS)). Pretreatment with L-NAME or AG enhanced the cumulative sodium excretion (CuU(Na)V) after 40 min VE in the innervated kidneys by 27 and 23 % (both P < 0.001), respectively, compared to the untreated control group, whereas they were without effect in the denervated kidneys. Cumulative urine flow (CuUV) after VE in L-NAME- and AG-treated groups was enhanced in both kidneys, by some 17-21 % in the denervated (P < 0.01) and 37 39 % in the innervated kidneys (P < 0.001) by comparison with the corresponding untreated controls. 7-NI had no effect on CuUV, but reduced CuU(Na)V in the denervated kidneys by 25 % (P < 0.01) when compared to the control group. The results suggested that NO, possibly generated by endothelial NOS (eNOS) and iNOS, was a contributory factor in mediating the renal response to VE. There appeared to be a tonic inhibitory action of NO on water excretion which was renal nerve independent, whereas its impact on sodium handling appeared to be dependent upon a background level of renal nerve activity. Experimental Physiology (2001) 86.1, 47-54. PMID- 11429620 TI - Re-expression of pulmonary surfactant proteins following tracheal obstruction in fetal sheep. AB - Increased fetal lung expansion, induced by tracheal obstruction (TO), is a potent stimulus for fetal lung growth, but rapidly reduces surfactant protein (SP) mRNA levels. Our aim was to determine the time course for the re-expression of the surfactant proteins in fetal lung tissue following the release of a TO and to relate these to the changes in lung liquid volume. Fetal sheep were exposed to either: (1) no treatment (controls); (2) 4 days of TO; (3) 4 days of TO, followed by release of the obstruction for 24 h; (4) 4 days of TO followed by release of the obstruction for 3 days. Four days of TO increased lung liquid volumes from 26.8 +/- 1.9 to 72.0 +/- 5.6 ml kg(-1) and reduced SP-A, SP-B and SP-C mRNA levels to 38.5 +/- 10.7, 56.8 +/- 10.3 and 18.3 +/- 5.3 % of control values, respectively. One day after TO release, lung liquid volumes were reduced to 17.4 +/- 5.3 ml kg(-1) (control 128 days, 31.0 +/- 3.8 ml kg(-1)) and SP-A and SP-B mRNA levels were not different from control levels. In contrast, SP-C mRNA levels only increased to 45.4 +/- 17.3 % of control. Three days after TO release, lung liquid volumes increased to 48.0 +/- 8.5 ml kg(-1) and SP-A and SP-B mRNA levels were reduced to 48.8 +/- 10.2 % and 71.5 +/- 19.8 % of control, respectively; SP C mRNA levels remained at 35.3 +/- 12.3 % of control. Following the release of a TO, SP-A, SP-B and SP-C mRNA levels were closely and inversely related to the volume of lung liquid. Based on these relationships, the lung liquid volumes that equate to 100 % expression were considerably less than control lung volumes (< 10 vs. 30-40 ml kg(-1)) in fetuses of this age. Thus, the changes in fetal lung SP A, SP-B and SP-C mRNA levels following the release of a TO are variable, differ between the proteins and are closely related to the changes in lung liquid volumes. We conclude that the re-expression of surfactant proteins following TO is variable and that the change in lung liquid volume is potentially a good indicator for surfactant protein re-expression. Experimental Physiology (2001) 86.1, 55-63. PMID- 11429621 TI - Effect of postnatal age and a beta(3)-adrenergic agonist (Zeneca D7114) administration on uncoupling protein-1 abundance in the lamb. AB - We examined the effect of time after birth and beta(3)-adrenergic agonist (Zeneca D7114) administration on uncoupling protein-1 (UCP1) abundance and thermoregulation in the lamb. Forty twin lambs, all born normally at term, were maintained at a cold ambient temperature of between 3 and 8 degrees C. At 0.5, 1.75, 5.25, 11.25 and 23.25 h after birth eight sets of twins were fed 20 ml of formula milk +/- 10 mg kg(-1) of beta(3)-adrenergic agonist, and 45 min after feeding brown adipose tissue (BAT) was sampled. Colonic temperature was measured and BAT analysed for UCP1 abundance, GDP-binding to mitochondrial protein (i.e. thermogenic activity) and catecholamine content. Colonic temperature declined between 1.25 and 6 h from 40.2 degrees C to 39.2 degrees C and then increased to 39.8 degrees C at 12 h, but increased after feeding at all ages. UCP1 abundance increased from 1.25 h after birth, to peak at 2 h after birth in controls, compared with 6 h after birth in beta(3)-adrenergic agonist-treated lambs. The level of GDP-binding to mitochondrial protein did not change significantly with age but was increased by beta(3)-adrenergic agonist treatment. The noradrenaline (norepinephrine) content of BAT increased between 1.25 and 12 h after birth, irrespective of beta(3)-adrenergic agonist administration. The total weight of perirenal BAT plus its lipid, protein and mitochondrial protein content declined over the first 6 h of life. UCP1 development continues over the first 24 h of neonatal life, and can be manipulated by beta(3)-adrenergic agonist administration. This may represent one method of improving thermoregulation in newborn lambs. Experimental Physiology (2001) 86.1, 65-70. PMID- 11429622 TI - Effects of gestation on ovine fetal and maternal angiotensin receptor subtypes in the heart and major blood vessels. AB - Previous studies in fetal sheep have concluded that (a) the vascular AT(1) angiotensin II (Ang II) receptor subtype is present in the external umbilical artery, but not in other systemic blood vessels, and (b) carotid arterial rings contract in vitro in response to Ang II. These contractions are blocked by the AT(1) specific receptor antagonist losartan. The aim of the present study was to resolve the apparent contradiction of these earlier conclusions, by examining the distribution of Ang II receptor subtypes in different regions of the ovine fetal cardiovascular system, and to find out at what stage in development AT(1) receptors first appear. We measured AT(1) and AT(2) receptors in hearts, carotid arteries, aortae and umbilical vessels from fetal sheep aged 65-144 days (term approximately 150 days), and in hearts and aortae from lambs, and adult pregnant and non-pregnant ewes. Both AT(1) and AT(2) receptors were present in aortae of fetuses > 118 days gestation, and carotid arteries of fetuses > 121 days gestation, while in younger fetuses only AT(2) receptors were found. The proportion of carotid artery and aortic AT(1) receptors increased with age, while the proportion of AT(2) receptors decreased. The internal umbilical artery contained both subtypes, but there was no relationship between receptor density and gestational age. The external umbilical artery had only AT(1) receptors. The highest density of Ang II receptors was found in the fetal heart where the AT(2) subtype predominated. The density of fetal cardiac Ang II receptors declined with age (r = -0.44, P < 0.02) due to the decrease in the AT(2) subtype. The density in late gestation fetal hearts was greater than in lamb or adult hearts (P < 0.001). Our study shows that fetal systemic blood vessels contain AT(1) receptors, and we have documented for the first time that the appearance of AT(1) receptors is both different in different regions of the fetal cardiovascular system and is developmentally regulated. Together with the in vitro contractile studies, this suggests that Ang II can play an important role in fetal blood pressure regulation via AT(1) receptors in the fetal systemic vasculature, as well in the umbilicoplacental vessels. Experimental Physiology (2001) 86.1, 71 82. PMID- 11429623 TI - Effects of exposure to high temperature and feeding level on regional blood flow and oxidative capacity of tissues in piglets. AB - To determine to what extent exposure to high ambient temperature and feeding level affect tissue energy metabolism in piglets, regional blood flow and oxidative capacity of tissues were evaluated in sixteen 21.8 +/- 2.8 kg pigs. At 5 weeks of age, littermates were divided into three groups and acclimated to the treatment for 25 days. One group was reared at 33 degrees C and fed ad libitum (33AL, n = 6) while the other two groups were maintained at 23 degrees C and either pair-fed on the basis of the food consumption of their 33AL littermates (23PF, n = 5), or fed ad libitum (23AL, n = 5). Regional blood flow was determined in conscious pigs by injection of coloured microspheres, which were recovered in different tissues after slaughter. Activities of cytochrome oxidase and cytochrome aa(3) content were measured in tissue homogenates of heart, longissimus dorsi and rhomboideus muscles, liver and small intestine. There was decreased blood flow to internal adipose tissue (42 %) and increased blood flow to peripheral tissues (skin, 44 %) and tissues implicated in respiratory evaporative heat loss (diaphragm, 45 %, lungs, 59 %) at 33 degrees C compared to 23 degrees C, which can be viewed as an effective mechanism for increasing heat loss at high temperature. In addition, the concomitant decrease in blood flow (49 %) and slight reduction of oxidative capacities in both muscles at 33 degrees C might contribute to the reduction in thermogenesis, but these effects were also observed when the feeding level was reduced at thermal neutrality (23PF group). In the viscera (intestine, liver), blood flow was decreased in the two groups on a restricted food intake (about 50 % of 23AL), independently of environmental temperature. The results suggest that most of the mechanisms associated with the reduction in energy expenditure during warm acclimation are related to the adaptive reduction in food intake. Experimental Physiology (2001) 86.1, 83-91. PMID- 11429624 TI - The contribution of chemoreflex drives to resting breathing in man. AB - The contribution of automatic drives to breathing at rest, relative to behavioural drives such as "wakefulness", has been a subject of debate. We measured the combined central and peripheral chemoreflex contribution to resting ventilation using a modified rebreathing method that included a prior hyperventilation and addition of oxygen to maintain isoxia at a P(ET,O2) (end tidal partial pressure of oxygen) of 100 mmHg. During rebreathing, ventilation was unrelated to P(ET,CO2) (end-tidal partial pressure of carbon dioxide) in the hypocapnic range, but after a threshold P(ET,CO2) was exceeded, ventilation increased linearly with P(ET,CO2). We considered the sub-threshold ventilation to be an estimate of the behavioural drives to breathe (mean +/- S.E.M. = 3.1 +/- 0.5 l min(-1)), and compared it to ventilation at rest (mean +/- S.E.M. = 9.1 +/- 0.7 l min(-1)). The difference was significant (Student's paired t test, P < 0.001). We also considered the threshold P(CO2) observed during rebreathing to be an estimate of the chemoreflex threshold at rest (mean +/- S.E.M. = 42.0 +/- 0.5 mmHg). However, P(ET,CO2) during rebreathing estimates mixed venous or tissue P(CO2), whereas the resting P(ET,CO2) during resting breathing estimates P(a,CO2) (arterial partial pressure of carbon dioxide). The chemoreflex threshold measured during rebreathing was therefore reduced by the difference in P(ET,CO2) at rest and at the start of rebreathing (the plateau estimates the mixed venous P(CO2) at rest) in order to make comparisons. The corrected chemoreflex thresholds (mean +/ S.E.M. = 26.0 +/- 0.9 mmHg) were significantly less (paired Student's t test, P < 0.001) than the resting P(ET,CO2) values (mean +/- S.E.M. = 34.3 +/- 0.5 mmHg). We conclude that both the behavioural and chemoreflex drives contribute to resting ventilation. Experimental Physiology (2001) 86.1, 109-116. PMID- 11429625 TI - Dynamic ventilatory response to acute isocapnic hypoxia in septuagenarians. AB - This study compared the ventilatory response to 20 min of acute isocapnic hypoxia (end-tidal P(O(2)), 50 mmHg) using the technique of dynamic end-tidal forcing in young (Y) and old (O) men. Two groups of non-smoking male subjects (mean +/- s.d. age: Y, 29.8 +/- 6.9 years; O, 73.4 +/- 2.8 years) with similar body size, normal age-predicted spirometry, and normal moderate levels of physical activity were studied. Compared with baseline ventilation in euoxia (10.79 +/- 1.99 and 11.88 +/- 0.91 l min-1) both groups responded to the abrupt onset of isocapnic hypoxia with peak ventilatory responses of 22.58 +/- 2.60 and 24.56 +/- 2.54 l min-1 for Y and O, respectively (not significant, n.s.). Both groups demonstrated a significant increment in neuromuscular drive (i.e. tidal volume (V(T))/inspiratory time (T(I)); 0.46 +/- 0.06 to 0.91 +/- 0.15 and 0.48 +/- 0.06 to 0.91 +/- 0.12 l s-1 for Y and O, respectively) with a small (but also significant) change in central timing (T(I)/total ventilation time (T(tot)); 0.38 +/- 0.02 to 0.41 +/- 0.02 and 0.42 +/- 0.02 to 0.45 +/- 0.02 for Y and O, respectively). Oxygen sensitivity was assessed using Weil's equation, and gave a hyperbolic factor (A) of 282 +/- 75 and 317 +/- 72, and using the linear equation: change in expiratory minute volume (DeltaV.(E))/change in arterial O(2) saturation (DeltaS(a,O(2))) which gave -1.17 +/- 0.57 and -1.17 +/- 0.42 l min-1 %-1 (n.s.) for Y and O, respectively. After 20 min of sustained isocapnic hypoxia, ventilation declined to 14.29 +/- 1.92 and 16.85 +/- 2.34 l min-1 for Y and O, respectively (n.s.). The acute response to hypoxia was characterised by similar time constants (16.0 +/- 5.4 and 18.5 +/- 6.7 s) and time delays (4.8 +/- 2.1 and 4.6 +/- 1.9 s) for Y and O, respectively. Thus, the dynamic ventilatory response to acute isocapnic hypoxia is maintained into the eighth decade in a group of habitually active elderly men. Experimental Physiology (2001) 86.1, 117 126. PMID- 11429626 TI - Corticospinal facilitation studied during voluntary contraction of human abdominal muscles. AB - Transcranial magnetic stimulation (TMS) of the human motor cortex was used to study facilitation of motor-evoked potentials (MEPs) in the rectus abdominis (RA) muscle, a trunk flexor, during voluntary activation. MEPs could be produced in the relaxed RA muscles of all six normal subjects studied. The MEPs had short latencies (18-22 ms) which are consistent with other studies suggesting a fast corticospinal input to the trunk muscles. Marked facilitation was observed in the MEPs when subjects were asked to produce graded levels of voluntary contractions. The two tasks used to produce voluntary contractions were a forced expiration during a breath-holding task (FEBH) and bilateral trunk flexion (BTF). Maximal voluntary EMG activity during the BTF task produced around 4.2 times more integrated EMG than during the FEBH task. Similarly the MEP amplitude at MVC was 2.3 times greater during BTF than FEBH. The pattern of MEP facilitation with increasing voluntary EMG was not linear and a maximal MEP amplitude was observed at a level of voluntary contraction around 30 % MVC in both tasks. There were some subtle differences in the pattern of facilitation in the two tasks. When TMS was applied to the right cortex only, MEPs were seen in both left and right RA muscles suggesting some ipsilateral corticospinal innervation. The latency of the right (ipsilateral) response was approximately 2 ms longer than the left. Comparison with studies in hand and leg muscles suggests that the facilitation pattern in RA may reflect a substantial degree of corticospinal innervation. Experimental Physiology (2001) 86.1, 131-136. PMID- 11429627 TI - Effect of caffeine co-ingested with carbohydrate or fat on metabolism and performance in endurance-trained men. AB - We examined the effect of caffeine co-ingested with either carbohydrate or fat on metabolism and performance in eight endurance-trained subjects who performed a random order of four experimental trials consisting of 120 min of steady-state ergometer cycling at 70 % of maximal O(2) uptake (SS) followed by a time trial in which subjects completed a set amount of work (7 kJ kg-1) as quickly as possible. One hour before SS subjects ingested either 2.6 g kg-1 carbohydrate (CHO); 2.6 g kg-1 CHO + 6 mg kg-1 caffeine (CHO + CAF); 1.2 g kg-1 fat with 2000 U I.V. heparin (FAT); or 1.2 g kg-1 fat with 2000 U I.V. heparin + 6 mg kg-1 caffeine (FAT + CAF). The rate of carbohydrate oxidation was higher (micromol kg-1 min-1: CHO, 243 +/- 39 and CHO + CAF, 239 +/- 30 vs. FAT, 196 +/- 48 and FAT + CAF, 191 +/- 55; P < 0.05, values are means +/- S.D.) and the rate of fat oxidation lower (micromol kg-1 min-1: CHO, 19 +/- 8 and CHO + CAF, 22 +/- 7 vs. FAT, 35 +/- 19 and FAT + CAF, 37 +/- 17; P < 0.05) with carbohydrate than fat ingestion. Yet despite lower carbohydrate use with fat feeding, the time taken to complete the time trial was less after carbohydrate than after fat ingestion (min: CHO, 30.37 +/- 7.42 and CHO + CAF, 29.12 +/- 5.62 vs. FAT, 33.02 +/- 8.50 and FAT + CAF, 32.78 +/- 7.70; P < 0.05). We conclude that (1) caffeine co-ingested with either carbohydrate or fat meals has no additive effect on substrate utilization or exercise performance and (2) carbohydrate ingestion before exercise improves subsequent time trial performance compared with fat ingestion. Experimental Physiology (2001) 86.1, 137-144. PMID- 11429628 TI - Functional characterisation of the volume-sensitive anion channel in rat pancreatic beta-cells. AB - The whole-cell and perforated patch configurations of the patch-clamp technique were used to characterise the volume-sensitive anion channel in rat pancreatic beta-cells. The channel showed high permeability (P ) relative to Cl(-) to extracellular monovalent organic anions (P(SCN)/P(Cll) = 1.73, P(acetate)/P(Cll) = 0.39, P(lactate)/P(Cll) = 0.38, P(acetoacetate)/P(Cll) = 0.32, P(glutamate)/P(Cll) = 0.28) but was less permeable to the divalent anion malate (P(malate)/P(Cll) = 0.14). Channel activity was inhibited by a number of putative anion channel inhibitors, including extracellular ATP (10 mM), 1,9 dideoxyforskolin (100 microM) and 4-OH tamoxifen (10 microM). Inclusion of the catalytic subunit of protein kinase A in the pipette solution did not activate the volume-sensitive anion channel in non-swollen cells. Furthermore, addition of 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP) or forskolin failed to activate the channel in intact cells under perforated patch conditions. Addition of phorbol 12,13-dibutyrate (200 nM), either before or after cell swelling, also failed to affect channel activation. Our findings do not support the suggestion that the volume-sensitive anion channel in pancreatic beta-cells can be activated by protein kinase A. Furthermore, the beta-cell channel does not appear to be subject to regulation via protein kinase C. Experimental Physiology (2001) 86.2, 145-150. PMID- 11429629 TI - Role of Ca(2+)-activated Cl(-) current in ventricular action potentials of sheep during adrenoceptor stimulation. AB - Adrenoceptor stimulation enhances repolarising and depolarising membrane currents to different extents in cardiac myocytes. We investigated the opposing effects of the repolarising Ca(2+)-activated Cl(-) current (I(Cl(Ca))) and depolarising L type Ca(2+) current (I(Ca,L)) on the action potential configuration of sheep ventricular myocytes stimulated with noradrenaline. Whole-cell current-clamp recordings revealed that noradrenaline accelerated and prolonged phase-1 repolarisation. We define the minimal potential at the end of phase-1 repolarisation as "notch level". Noradrenaline (1 microM) caused the notch level to fall from 14 +/- 2.6 to 7.8 +/- 2.8 mV (n = 24), but left action potential duration, resting membrane potential or action potential amplitude unaffected. Whole-cell voltage-clamp recordings showed that 1 microM noradrenaline increased both I(Ca,L) and I(Cl(Ca)), but it had no significant effect on the principal K(+) currents. Blockage of I(Cl(Ca)) by 0.5 mM 4,4'-diisothiocyanatostilbene-2,2' disulphonic acid (DIDS) in both the absence and the presence of noradrenaline abolished phase-1 repolarisation. In the presence of noradrenaline, DIDS caused elevation of the plateau phase amplitude and an increase in the action potential duration. In conclusion, elevation of the plateau phase amplitude and action potential prolongation associated with an increased I(Ca,L) upon adrenoceptor stimulation is prevented by an increased I(Cl(Ca)) in sheep ventricular myocytes. Experimental Physiology (2001) 86.2, 151-159. PMID- 11429630 TI - Effects of reactive oxygen species on aspects of excitation-contraction coupling in chemically skinned rabbit diaphragm muscle fibres. AB - Oxidants have been suggested to enhance contractile function in unfatigued muscle. In this study we aimed to determine the effect of oxidants on "chemically skinned" diaphragm muscle fibre bundles. The sarcoplasmic reticulum and contractile proteins were exposed to superoxide anions (O2-) and hydrogen peroxide (H2O2) under controlled conditions. Application of O2-initially increased maximum Ca2+ -activated force but subsequently reduced maximum Ca2+ activated force without altering myofilament Ca2+ sensitivity. Unlike myocardium, caffeine-induced Ca2+ release from the sarcoplasmic reticulum was also inhibited by O2- exposure in diaphragm fibre bundles. Application of H2O2 also increased maximum Ca2+ -activated force but had additional effects on resting tension (which increased to 25 % of the control maximum Ca2+ -activated force). H2O2 was without effect on myofilament Ca2+ sensitivity or caffeine-induced Ca2+ release from the sarcoplasmic reticulum. These data demonstrate that oxidants can potentiate contractile force in the diaphragm through a direct action on the contractile proteins. The potentiation of force is not sustained, however, and under these conditions the detrimental effects of O2- on Ca2+ release from the sarcoplasmic reticulum combined with the effects of oxidants on the contractile proteins will ultimately compromise excitation-contraction coupling in the diaphragm. Experimental Physiology (2001) 86.2, 161-168. PMID- 11429631 TI - Effects of T-lymphocyte-dependent and -independent immunity on cholinergic enzyme activity in mouse lacrimal gland. AB - The nature of the immune response following ocular immunization with a protein and a polysaccharide and the effects such immunization have on the activities of cholinergic enzymes in the lacrimal glands of BALB/c mice were examined. Lacrimal glands are highly innervated by sympathetic and parasympathetic nerve fibres and are involved in mucosal immunity and therefore are excellent sites to study neuro immune interactions. In this report, a T-lymphocyte-dependent protein antigen, keyhole limpet haemocyanin (KLH) and a T-lymphocyte-independent polysaccharide antigen, dextran (DEX) were administered topically to the eyes or intraperitoneally injected. Both routes of immunization produced a strong serum antibody response when KLH was the antigen. DEX, however, evoked a serum antibody response only after intraperitoneal administration. Eosin-haematoxylin staining indicated no histological abnormality or inflammatory changes in any immunized lacrimal glands, but immuno-staining revealed that only in the KLH-treated tissues were IgG-producing plasma cells discernible. Furthermore, KLH-specific antibody was also detectable using an immuno-blot assay in lacrimal glands. Polymerase chain reaction analysis with cytokine-specific primers revealed induction of interleukin-4 (lL-4) in KLH-treated lacrimal glands, but not in DEX or unimmunized tissues. Thus, the nature of the antigen seems important in the induction of the immune response in lacrimal glands. To delineate the effects that immunogenic differences might have on the activities of the cholinergic enzymes, choline acetyl-transferase (ChAT) and acetylcholinesterase (AChE) were assayed using radiolabelled substrates and measuring labelled products. Both ChAT and AChE activities were influenced following KLH immunization, while DEX had only transient effects on ChAT. This is possibly due to the fact that KLH, a protein antigen, is the effective inducer of the specific immune response in the lacrimal gland, while DEX is not. Experimental Physiology (2001) 86.2, 169-176. PMID- 11429632 TI - Pacemaker shift in the rabbit sinoatrial node in response to vagal nerve stimulation. AB - Effects of brief postganglionic vagal nerve stimulation on the activation sequence of the rabbit sinoatrial (SA) node were investigated. Activation sequences in a small area (7 mm x 7 mm) on the epicardial surface were measured in a beat-to-beat manner using an extracellular potential mapping system composed of 64 modified bipolar electrodes with high-gain and low-frequency band-pass filtering. The leading pacemaker site was recognised clearly from both the activation sequence and the characteristic morphology of the potentials. Vagal stimulation resulted in a short-lasting initial slowing of spontaneous rate followed by a long-lasting secondary slowing; a brief period of relative or absolute acceleration was interposed between the two slowing phases. During these changes of spontaneous rate, the leading pacemaker site shifted in a complex beat to-beat manner by 1-6 mm alongside the crista terminalis in the superior or inferior direction. For the first spontaneous excitation following stimulation, the greater the slowing, the larger the distance of the pacemaker shift. There was no such linear relationship between the extent of slowing and the distance of pacemaker shift for the subsequent beats. These changes in the leading pacemaker site in response to vagal stimulation may be the result of the functional and morphological heterogeneity of the mammalian SA node in terms of innervation, receptor distribution and ion channel densities. Experimental Physiology (2001) 86.2, 177-184. PMID- 11429633 TI - The effects of dietary creatine supplements on the contractile properties of rat soleus and extensor digitorum longus muscles. AB - Daily creatine supplements (0.258 g kg(-1) ) were administered to adult male Wistar rats (n = 7) in the drinking water. Age matched rats (n = 6) acted as controls. After 5-6 days, contractile properties were examined in soleus and extensor digitorum longus (EDL) muscle strips in vitro at 30 degrees C. In soleus muscles, creatine supplements decreased the half-relaxation time of the isometric twitch from 53.6 +/- 4.3 ms in control muscles to 48.4 +/- 5.5 ms but had no effect on twitch or tetanic tension or on twitch contraction time. In EDL muscles twitch tension, tetanic tension, twitch contraction and half-relaxation times were all unaffected by creatine supplements. Creatine supplements increased the fatigue resistance of the soleus muscles but had no effect on that of the EDL muscles. After a 5 min low-frequency fatigue test, tension (expressed as a percentage of initial tension) was 56 +/- 3 % in control soleus muscles, whereas that in the creatine-supplemented muscles was 78 +/- 6 % (P < 0.01). In the EDL muscles, the corresponding values were 40 +/- 2 % and 41 +/- 9 %, respectively. The force potentiation which occurred in the EDL muscles during the initial 20-30 s of the fatigue test was 170 +/- 10 % of initial tension in the control muscles 24 s after the initial stimulus train but was reduced (P < 0.01) to 130 +/- 20 % in the creatine-supplemented muscles. In conclusion, soleus muscle endurance was increased by creatine supplements. EDL endurance was unaffected but force potentiation during repetitive stimulation was decreased. Experimental Physiology (2001) 86.2, 185-190. PMID- 11429634 TI - Expression of constitutive but not inducible cyclooxygenase maintains articular perfusion in the rat knee. AB - Experiments were performed in the normal rat knee joint to investigate the role of different isoforms of cyclooxygenase (COX) in the regulation of basal joint blood flow. Laser Doppler imaging (LDI) was used to measure articular perfusion, and reverse transcriptase polymerase chain reaction (RT-PCR) for the detection of COX-1 and COX-2 mRNA in joint tissue. Intravenous infusion of indomethacin (a non selective inhibitor of COX; 0.34 nmol min(-1)) over 40 min produced a time dependent increase in articular vascular resistance (maximum 22.5 % at 40 min; P < 0.0001, one-way ANOVA) whereas vehicle over a similar time period had no effect in a control group. An equimolar concentration of a highly selective inhibitor for COX-2, SC-236, was administered in a further group of rats but this did not increase articular vascular resistance. While there was no significant difference between the response to vehicle and SC-236 (two-way ANOVA; P = 0.686, n = 6) the response to indomethacin was significantly greater than vehicle or SC-236 (two way Anova; P < 0.0001, n = 6). COX-1, but not COX-2, was detectable by RT-PCR in all joint tissue samples examined (n = 4). The results of this study indicate that prostaglandins (PGs) play an important role in the maintenance of basal perfusion in the rat knee joint, with COX-1 being the physiologically relevant isoform. Experimental Physiology (2001) 86.2, 191-197. PMID- 11429635 TI - Feto-maternal relationships in goats during heat and cold exposure. AB - Maternal and fetal body temperatures were measured in five Boer goats, of mean mass 64 +/- 8 kg, using temperature-sensitive radiotelemeters implanted intra abdominally. Body temperatures were recorded every 5 min. Throughout the last month of gestation, fetal temperature was approximately 0.6 (o)C higher than that of the mother, in normal laboratory conditions (ambient air temperature: 21-24 (o)C). This feto-maternal temperature difference between the goat fetus and its mother is similar to that found in other mammals, including sheep. When the pregnant goats were subjected to short-term heating and cooling, the difference between maternal and fetal body temperatures changed. Thus the mean difference between fetal and maternal body temperatures decreased from 0.4 to 0.2 (o)C during 2 h of heating, while it widened from 0.3 to 0.7 (o)C during 6 h of cooling. These data support the idea that the fetus is thermally protected from excursions of body temperature during changes in the mother's thermal environment. Reports of goat stock losses and abortions during cold spells in their natural habitats may be the result of more severe and/or prolonged cold exposures that not only adversely affect fetal or maternal body temperature, but also influence other aspects of metabolism. Experimental Physiology (2001) 86.2, 199-204. PMID- 11429636 TI - Altered glycogen synthase and phosphorylase activities in skeletal muscle of tetraplegic patients. AB - Despite marked differences in both the extent of physical activity and in muscle metabolism and structure between tetraplegic and control subjects, the glycogen content in the skeletal muscle of both groups is similar. We determined whether this similarity could be explained by the activities of key enzymes of glycogen metabolism. Muscle biopsies were analysed for glycogen synthase (GS) and glycogen phosphorylase (GP) activities, as well as for metabolites. Glycogen content did not differ significantly between the two groups. Total glycogen synthase activity was reduced by almost 60 % in tetraplegics (P < 0.01), whereas total phosphorylase activity did not differ between groups. GS fractional activity did not differ between groups, whereas phosphorylase fractional activity (-/+ AMP) was significantly higher in the tetraplegics (0.08 +/- 0.01, control; 0.25 +/- 0.02, tetraplegics; P < 0.001). Neither uridine diphosphate (UDP)-glucose nor glucose 6-phosphate (G-6-P) content in muscle differed significantly between groups. These data demonstrate that, in tetraplegics, muscle glycogen content is preserved despite decreases in GS activity and increases in phosphorylase fractional activity. Muscle paralysis has differential effects on the activities of GS and GP. Experimental Physiology (2001) 86.2, 205-209. PMID- 11429638 TI - Overview of current research in parturition. AB - The mechanism of human parturition is not understood and further research into this important physiological process is needed. Preterm labour remains a major cause of perinatal mortality and morbidity and there is controversy about the effectiveness of current tocolytic agents. In some species, notably the sheep, parturition is preceded by an activation of the fetal hypothalamic-pituitary adrenal axis. However, in primates this axis has a supportive, rather than essential, role. A fall in maternal progesterone levels is a prerequisite for parturition in most mammals and this takes place either through increased conversion of progesterone to oestrogens in the placenta, or through the demise of the corpus luteum of pregnancy, depending on the species. In primates and guinea-pigs parturition occurs without an apparent fall in maternal progesterone levels. Gene targeting experiments in mice have demonstrated the critical role of prostaglandin FP receptors, necessary to mediate the luteolytic effect of PGF(2alpha) before parturition. Prostaglandin synthesis is required for the onset and progress of labour as demonstrated by experiments with cPLA(2)- and PGHS-1 deficient mice. The importance of local tissue conversion of progesterone to reduced androgens in the regulation of cervical ripening has been demonstrated in 5alpha-reductase-deficient mice. The chronic and ubiquitous gene inactivation obtained with conventional methods has disadvantages, in that it may allow the activation of compensating pathways, making the interpretation of results difficult. This problem may be overcome by using pulsed and tissue-selective gene knockout strategies. The study of human parturition is complicated by the lack of access to direct experimentation, whereas the endocrine differences between species make it difficult to extrapolate animal data to humans. However, the development of genomic/proteomic technologies that allow the simultaneous screening of thousands of genes and gene products in small samples of tissue, and new methods to study the biochemistry of receptors and proteins involved in smooth muscle physiology promise new insights into the control of human labour. Nevertheless, the integration of rapidly expanding knowledge into a complete understanding of the roles of the mother and the fetus in the initiation of parturition, and the development of selective medication for the effective management of preterm labour remain an arduous challenge for the next decade. Experimental Physiology (2001) 86.2, 213-222. PMID- 11429639 TI - Hormones and calcium: mechanisms controlling uterine smooth muscle contractile activity. The Litchfield Lecture. AB - The regulation of myometrial contraction is of paramount importance for the maintenance of pregnancy and for parturition. Understanding this regulation involves delineating the pathways that control myometrial contraction and relaxation and defining the regulation of these pathways. The pathways can be broken down further into those signalling cascades controlling the concentration of intracellular free calcium (Ca(2+)(i)) and those controlling the contractile apparatus itself. This discussion focuses primarily on the former and their regulation during pregnancy. In particular, cross-talk between the contractant and relaxant signalling pathways mediated through cyclic AMP is markedly changed at the end of pregnancy. Experimental Physiology (2001) 86.2, 223-237. PMID- 11429640 TI - The physiological basis of uterine contractility: a short review. AB - In this review we discuss our current understanding of the cellular basis of uterine contractility, highlighting those areas requiring further study. It is clear that the basic processes of excitation-contraction coupling lie within the myometrial cell, and that these may be modified by agonists. Pacemaker activity, however, remains a mystery. The contribution of extracellular calcium entry to contraction is shown to be vital, whilst the role of the sarcoplasmic reticulum remains controversial. Much current experimental focus is on pathways controlling and regulating contraction, and we discuss sensitisation mechanisms and question their role in intact uterine preparations. Experimental Physiology (2001) 86.2, 239-246. PMID- 11429641 TI - Regulation of human myometrial contractility during pregnancy and labour: are calcium homeostatic pathways important? AB - If we are to develop new strategies for the treatment and management of preterm and dysfunctional term labour, it is imperative that we improve current understanding of the control of human uterine activity. Despite many studies of animal pregnancy, there is a paucity of knowledge relating to the complex control of human myometrium during pregnancy. It is hypothesized that human myometrium is relatively quiescent during the majority of pregnancy and that as term approaches there is cascade of molecular events that prepare the uterus for labour. This review will consider the cellular mechanisms involved in the regulation of human myometrial activity and the modulation of these by hormonal and mechanical signals. In particular, the contribution of calcium homeostatic pathways to the control of human myometrial contractility during gestation will be discussed. Experimental Physiology (2001) 86.2, 247-254. PMID- 11429642 TI - Potassium channels in the human myometrium. AB - The contractility of the human uterus is under the fine control of a variety of interacting bioactive agents. During labour, the excitability of the uterus is drastically transformed in comparison with the non-labour state and is manifest at the membrane level via the activity of uterine ion channels. This article reviews the contribution of potassium (K(+)) channels to human uterine excitability. Experimental Physiology (2001) 86.2, 255-264. PMID- 11429643 TI - Uterine quiescence: the role of cyclic AMP. AB - It is accepted that whilst hormones such as oxytocin, vasopressin and prostaglandin F2alpha induce myometrial contractions, essentially via an elevation of intracellular calcium, other ligands, such as beta-adrenoceptor agonists, calcitonin gene-related peptide, and prostaglandin E2, promote uterine quiescence via their ability to increase intracellular cyclic AMP levels. At present, the exact factors initiating human parturition remain unknown, and labour may occur due to a loss of uterine quiescence, an increase in uterine contractility, or a combination of both. Whilst many studies have aimed to understand the mechanisms underlying uterine contractility there is a relative paucity of data regarding myometrial relaxation. We have verified the presence of mRNA encoding adenylyl cyclase (AC) isoforms I, II, III, V, VI, VII, VIII and IX in both non-pregnant and pregnant human myometrium, and in isolated myometrial cells maintained in cell culture. Furthermore, by means of immunoblotting and immunocytochemistry, we have demonstrated the expression of these isoforms as membrane-associated AC proteins, and identified changes in individual AC isoform expression during gestation. These findings illustrate the diversity of potential cAMP generating pathways in human myometrium, and the complexity of the signal transduction systems underlying uterine quiescence. Experimental Physiology (2001) 86.2, 265-272. PMID- 11429644 TI - Corticotrophin releasing hormone: its potential for a role in human myometrium. AB - Aside from its role as a hypothalamic stress hormone, corticotrophin releasing hormone (CRH) is also a placental hormone, at least in primates. Although the function of placentally derived CRH remains to be fully elucidated, elevated CRH levels have been associated with premature labour, suggesting that the hormone may be involved in regulating the duration of pregnancy. Indeed, pregnant human myometrium expresses functional CRH receptors (CRH R1 and CRH R2 subtypes) thought to signal predominantly via the second messenger cAMP. Thus, like other cAMP-producing hormones in the myometrium such as beta(2) agonists, CRH may play a part in maintaining uterine quiescence. However, several of the CRH receptor isoforms identified to date have a reduced ability to activate adenylate cyclase, raising the question as to whether they are linked to other signal transduction pathways. Here, we discuss critically the evidence for the peptide's role in regulating contractility, both directly at the myometrium and indirectly via the fetal membranes and decidua. The possibility of a role in myometrial growth modulation is also described. Experimental Physiology (2001) 86.2, 273-281. PMID- 11429645 TI - Receptor-coupled contractility of uterine smooth muscle: from membrane to myofilaments. AB - A comprehensive understanding of the mechanisms by which agonists control uterine contraction is essential for the successful clinical management of parturition and for the timely treatment of situations involving inappropriate uterine performance. In this review we discuss some of the key stimulatory mechanisms linking receptor occupation at the myometrial plasma membrane with alteration of myofilament activation. We focus on evidence that receptor-induced membranous recruitment of the small G-protein rhoA, and its downstream effector rho associated kinase (ROK) is crucial to agonist-induced Ca(2+)-sensitisation of uterine contraction and that co-ordination of this signal transduction pathway may be mediated by the actions of caveolins, proteins integral to specialised membranous regions termed caveolae. Experimental Physiology (2001) 86.2, 283-288. PMID- 11429646 TI - The structure and regulation of the oxytocin receptor. AB - The oxytocin receptor (OTR) is part of an ancient hormone system expressed in diverse phyla in relation to acute reproductive smooth muscle responses, such as egg-laying, birth, or milk letdown. The regulation of the OTR gene, while correlating with steroid levels in vivo, remains elusive. There appear to be both inhibitory and stimulatory influences acting upon a constitutive pattern of basal expression. We have found no evidence, however, for an effect of the sex steroids either directly on gene transcription, or on the receptor itself at the protein level. In the prostatic carcinoma cell line Du145, we have shown that up regulation of the OTR gene transcription can be effected by cAMP. In an attempt to characterize the expression of the OTR protein in vivo, we have shown, using ligand-blotting, that the OTR can be expressed at different sizes in transfected cells and in myometrium. Also, in the myometrium at term, immunohistochemistry suggests that there is both an increase in OTR protein per cell, as well as in the number of smooth muscle cells expressing OTR, emphasizing that perinatal changes are the results of both individual gene activation events and gross cellular differentiation. The OTR is a valuable model system reflecting molecular changes in the perinatal period. When we understand how this important molecule is regulated, we will also be a long way towards understanding the mechanisms controlling myometrial contractility at birth. Experimental Physiology (2001) 86.2, 289-296. PMID- 11429647 TI - Oxytocin antagonists: clinical and scientific considerations. AB - Preterm delivery is the largest cause of perinatal mortality and morbidity, yet the treatment of preterm labour has not been demonstrated to improve outcome. The reasons are numerous and complex, but they include a failure to understand the mechanism(s) of preterm labour, the multitude of different causes, the difficulty in diagnosis and the problems of outcome measurement in clinical trials. Recently, an oxytocin antagonist (atosiban) has been introduced into clinical practice in Europe. Although it may be an effective tocolytic, a beneficial effect on perinatal outcome has not been demonstrated. Atosiban has an effect at both oxytocin and vasopressin (V(1a)) receptors, which (assuming efficacy) raises the question as to whether oxytocin or vasopressin V(1a) antagonism is required for tocolysis. This review examines the rationale for tocolysis in preterm labour, the evidence for administration of atosiban and the role for oxytocin, vasopressin and their receptors in the onset of labour. Experimental Physiology (2001) 86.2, 297-302. PMID- 11429648 TI - Desensitisation of the oxytocin receptor and other G-protein coupled receptors in the human myometrium. AB - G-protein coupled receptors (GPCRs) are key to maintaining uterine quiescence and inducing phasic contractions at term. However, the biochemical mechanisms whereby uterine GPCRs are desensitised and re-sensitised during these physiological conditions are unknown. For example, the number of oxytocin receptors (OTRs) on uterine myocytes decrease significantly after the addition of oxytocin. Therefore, further understanding of the desensitisation/re-sensitisation of the OTR and other uterine GPCRs during pregnancy may provide a target for more efficient tocolytic drugs and more selective ways to modulate uterine activity. Here, we briefly review some of the mechanisms that may be involved during OTR and other GPCR desensitisation. Experimental Physiology (2001) 86.2, 303-312. PMID- 11429649 TI - Myosin light chain kinase and the onset of labour in humans. AB - Myosin light chain kinase (MLCK) is essential for myometrial contractions induced by calcium-mobilizing agonists. From the gene of vertebrate smooth muscle/non muscle MLCK there are at least four proteins expressed. We have found that both a > 200 and a 137 kDa MLCK are equally expressed in human non-pregnant (NP) and term pregnant (P) uterine smooth muscle and confirmed that 19 kDa telokin (TK) is only expressed in P myometrium. In addition, we have observed that a MLCK immunogen at approximately 60 kDa is only expressed in NP myometrium, suggesting that its expression is inhibited during normal pregnancy in a hormonally dependent manner. However, when we compared pregnant myometrium from patients delivered preterm (PT) (< 34 weeks gestation), but not in labour (NIL), with PT patients in labour (IL) we found that PT(IL) samples expressed the approximately 60 kDa MLCK immunogen and thus displayed a NP phenotype whereas PT(NIL) samples did not express the protein and retained a pregnant phenotype. We hypothesize that the novel approximately 60 kDa MLCK immunogen contributes to the aberrent contractility associated with preterm labour. Experimental Physiology (2001) 86.2, 313-318. PMID- 11429650 TI - Involvement of calmodulin in glucagon-like peptide 1(7-36) amide-induced inhibition of the ATP-sensitive K+ channel in mouse pancreatic beta-cells. AB - The present investigation was designed to examine whether calmodulin is involved in the inhibition of the ATP-sensitive K+ (K(ATP)) channel by glucagon-like peptide 1(7-36) amide (GLP-1) in mouse pancreatic beta-cells. Membrane potential, single channel and whole-cell currents through the K(ATP) channels, and intracellular free Ca2+ concentration ([Ca2+]i) were measured in single mouse pancreatic beta-cells. Whole-cell patch-clamp experiments with amphotericin perforated patches revealed that membrane conductance at around the resting potential is predominantly supplied by the K(ATP) channels in mouse pancreatic beta-cells. The addition of 20 nM GLP-1 in the presence of 5 mM glucose significantly reduced the membrane K(ATP) conductance, accompanied by membrane depolarization and the generation of electrical activity. A calmodulin inhibitor N-(6-aminohexyl)-5-chloro-1-naphthalenesulphonamide (W-7, 20 microM) completely reversed the inhibitory actions of GLP-1 on the membrane K(ATP) conductance and resultant membrane depolarization. Cell-attached patch recordings confirmed the inhibition of the K(ATP) channel activity by 20 nM GLP-1 and its restoration by 20 microM W-7 or 10 microM calmidazolium at the single channel level. Bath application of 20 microM W-7 also consistently abolished the GLP-1-evoked increase in [Ca2+]i in the presence of 5 mM glucose. These results strongly suggest that the mechanisms by which GLP-1 inhibits the K(ATP) channel activity accompanied by the initiation of electrical activity in mouse pancreatic beta cells include a calmodulin-dependent mechanism in addition to the well-documented activation of the cyclic AMP-protein kinase A system. PMID- 11429651 TI - Effects of prolonged aerobic exercise on myocardial responses to ischaemia reperfusion in the rat. AB - The effects of low-intensity, prolonged swimming on functional recovery of the rat heart (Langendorff preparations) from ischaemia-reperfusion (I/R) were investigated. Three groups of rats (120 days old) were used: sedentary rats (S) and rats exercised by a single bout of swimming lasting 5 (E5) or 8 h (E8), respectively. The effect of exercise on the response to I/R was related to an index of oxidative damage such as lipid peroxidation, as well as to the tissue antioxidant capacity and the response of heart tissue to in vitro oxidative stress. The intrinsic performance of E5 Langendorff preparations paced at 220 beats x min(-1) was also determined. A group of sedentary animals was used for H2O2-treated preparations. The effect of antioxidant treatment on inotropic recovery during reperfusion was studied on preparations from 5 or 8 h swimming vitamin E-treated (EVT5 and EVT8 and 5 or 8 h swimming untreated (EVU5 and EVU8) rats. Hearts from exercised animals displayed a reduced preischaemic inotropism, which in E5 rats was accompanied by an increase in the intrinsic heart rate. The lower intrinsic cardiac inotropism of E5 animals was confirmed in the paced preparations. The reduced contractility found in control hearts after addition of H2O2 to perfusion medium suggested that the low inotropism of E5 and E8 hearts was due to an exercise-induced increase in reactive oxygen species. Inotropic recovery during reperfusion was low in the S hearts, was significantly increased in the E5 hearts, and was again reduced to the S level in the E8 hearts. In the E8 hearts the indexes of cellular damage (LDH release) and oxidative stress increased, and antioxidant capacity decreased, while in E5 hearts there was no evidence of significant changes in such parameters. Performance and reperfusion recovery of hearts from 5 h swimming rats was not affected by vitamin E treatment, while those of hearts from 8 h swimming rats was the highest observed. We suggest that the higher inotropic recovery during reperfusion in the hearts from the E5 rats is related to the negative inotropic effect of exercise. The fall in recovery following the 8 h exercise was instead related to the increased oxidative stress. PMID- 11429652 TI - Rat muscle microvascular PO2 kinetics during the exercise off-transient. AB - Dependent upon the relative speed of pulmonary oxygen consumption (VO2) and blood flow (Q) kinetics, the exercise off-transient may represent a condition of sub- or supra-optimal perfusion. To date, there are no direct measurements of the dynamics of the VO2/Q relationship within the muscle at the onset of the work/recovery transition. To address this issue, microvascular PO2 (PO2,m) dynamics were studied in the spinotrapezius muscles of 11 female Sprague-Dawley rats (weight approximately 220 g) during and following electrical stimulation (1 Hz) to assess the adequacy of Q. relative to VO2 post exercise. The exercise blood flow response (radioactive microspheres: muscle Q increased approximately 240 %), and post-exercise arterial blood pH (7.40 +/- 0.02) and blood lactate (1.3 +/- 0.4 mM x l(-1)) values were consistent with moderate-intensity exercise. Recovery PO2,m (i.e. off-transient) rose progressively until baseline values were achieved ((Delta)end-recovery exercise PO2,m, 14.0 +/- 1.9 Torr) and at no time fell below exercising PO2,m. The off-transient PO2,m was well fitted by a dual exponential model with both fast (tau = 25.4 +/- 5.1 s) and slow (tau = 71.2 +/- 34.2 s) components. Furthermore, there was a pronounced delay (54.9 +/- 10.7 s) before the onset of the slow component. These data, obtained at the muscle microvascular level, support the notion that muscle VO2 falls with faster kinetics than muscle Q during the off-transient, such that PO2,m increases systematically, though biphasically, during recovery. PMID- 11429653 TI - Reflex nature of the cardiorespiratory response to primary thoracic blast injury in the anaesthetised rat. AB - Blast injuries represent a problem for civilian and military populations. Primary thoracic blast injury causes a triad of bradycardia, hypotension and apnoea. The objective of this study was to investigate the reflex nature of this response and its modulation by vagotomy or administration of atropine. The study was conducted on terminally anaesthetised (alphadolone/alphaxalone, 18-24 mg x kg x h(-1), I.V.) male Wistar rats randomly allocated to the groups indicated below. Blast injuries were produced with compressed air while sham blast involved the sound of a blast only. Primary blast injury to the thorax resulted in a bradycardia (measured as an increase in the interval between beats, or heart period (HP) to 489 +/- 37 ms from 133 +/- 3 ms with a latency of onset of 4.3 +/- 0.3 s, mean +/ S.E.M.), hypotension (fall in mean arterial blood pressure (MBP) from 128.1 +/- 3.7 mmHg to 34.8 +/- 4.1 mmHg, latency of onset 2.0 +/- 0.1 s) and apnoea lasting 28.3 +/- 2.3 s. Sham blast had no effect. The bradycardia and apnoea following thoracic blast were abolished by cervical vagotomy while the hypotension was attenuated. Atropine (0.3 mg x kg(-1), I.V.) caused a significant reduction in the bradycardia (HP increasing from 124 +/- 3 ms to 142 +/- 4 ms) but did not modulate either the hypotension or apnoea. It is concluded that a reflex involving the vagus nerve mediates the bradycardia, apnoea and a component of the hypotension associated with thoracic blast. The pattern of this response is similar to effects that follow stimulation of the pulmonary afferent C-fibres. PMID- 11429654 TI - Role of endothelin ET(A) receptor antagonism in the post-transplant renal response to angiotensin II in the rat. AB - The role of endothelins in the renal damage associated with ischaemic-reperfusion (I-R) injury during organ transplantation was determined by selective blockade of the ET(A) receptors with the receptor antagonist ABT-627. The integrity of kidney function was determined 2 and 8 weeks after transplantation by investigation of the renal response to angiotensin II. Under pentobarbitone anaesthesia (70 mg x kg(-1), I.P.), rats underwent a right nephrectomy. Transplantation of the left kidney was performed after 2 h cold ischaemia without or with ABT-627 treatment. Control animals underwent left renal denervation. The renal response to angiotensin II was measured 2 weeks later following blockade of endogenous production of angiotensin II with captopril. A further transplant group was allowed to recover for 8 weeks before the terminal study. In the control group, angiotensin II reduced renal blood flow (RBF), glomerular filtration rate (GFR), urine flow rate (UV), and fractional sodium excretion (FE(Na)) by 29 +/- 5 %, 19 +/- 4 %, 25 +/- 4 % and 32 +/- 7 %, respectively. Conversely, in the transplant group, angiotensin II left RBF unchanged and increased GFR (59 +/- 12 %) and UV (93 +/- 8 %). FE(Na) decreased by 24 +/- 9 %. In both the transplant group treated with ABT-627 and the long-term recovery group, the renal response to angiotensin II was normalised. In conclusion, renal transplantation following 2 h cold I-R injury resulted in a temporary abnormal renal response to angiotensin II, which was reversed by ET(A) receptor antagonism at the time of transplantation. PMID- 11429655 TI - Impact of the timing of indomethacin treatment in a model of synchronized bone remodelling in rats. AB - Prostaglandins (PGs) promote both bone resorption and formation in vitro and in vivo. In a synchronised model of bone remodelling, indomethacin, an inhibitor of PG synthesis, given from the start of the sequence, transiently impaired bone resorption. In this study we further explored the involvement of PGs in this model by treating rats with indomethacin (7.5 mg x kg(-1) x day(-1)) for 6 days from the peak of resorption (day 4 after activation in this model) or during reversal (day 6 after activation). In rats treated from day 4, the resorption surface (Oc.S/BS) and the number of osteoclasts (N.Oc/BPm) were higher on day 10 (+69 %, P < 0.01, and +60 %, P < 0.02 compared with controls, respectively); no effect on cell resorptive activity was observed. The bone formation surface (OS/BS) was reduced (-50 %, P < 0.01). The inactive surface (In/BS) was not modified. In rats treated from day 6, the Oc.S/BS was also higher than in controls (P < 0.02), as was the N.Oc/BPm (P < 0.05). Osteoclast activity appeared to be increased, as the osteoclast-bone interface was larger (P < 0.02), but the mean lacuna area was reduced (-23 %, P < 0.05). Bone formation was also strongly affected: the OS/BS was decreased (-66 %, P < 0.01), as was the osteoid seam thickness (-24 %, P < 0.05). The In/BS was increased 1.5-fold (P < 0.05). These data indicate that PGs intervene at various stages of this remodelling sequence, as both resorption and formation were affected by indomethacin. Although resorption resumed in the two treatment groups despite treatment continuation, the timing of treatment was clearly important. Only inhibition of PG synthesis at the peak of resorption delayed all phases of the remodelling sequence. In contrast, inhibition during the reversal phase prevented activation of a significant part of the bone surface usually involved at this stage of remodelling; this treatment schedule reduced the resorptive capacity of the system, and depressed osteoblast activity. PMID- 11429656 TI - Responses to stimulation of coronary and carotid baroreceptors and the coronary chemoreflex at different ventricular distending pressures in anaesthetised dogs. AB - Stimulation of left ventricular mechanoreceptors was believed not only to exert important effects on the circulation, but also to influence the responses to baroreceptor reflexes. However, most previous work is flawed due to inadequate localisation of stimuli to specific reflexogenic areas. In this study, we applied a discrete stimulus to left ventricular mechanoreceptors to examine other reflexes known to effect the circulation. Dogs were anaesthetised, artificially ventilated and a cardiopulmonary bypass established. The pressure distending the left ventricle was controlled through an apical cannula with the aortic valve obstructed by a balloon. Changes in ventricular systolic and end-diastolic pressure had only a small effect on vascular resistance, assessed as perfusion pressure in the systemic circulation (flow constant). Responses to changes in carotid or coronary pressure or to stimulation of chemosensitive afferents by injecting veratridine into the coronary circulation were always much larger. Responses to stimulation of these reflexes were little affected by the level of stimulus to the ventricular receptors. These experiments confirm that responses to stimulation of ventricular mechanoreceptors are very small and show that they remain small at different levels of input to other baroreceptive regions. There was no evidence of interaction between ventricular mechanoreceptor reflexes and carotid or coronary baroreceptors or ventricular chemosensitive reflexes. PMID- 11429657 TI - The independence of lung liquid absorption in postnatal sheep on pulmonary blood flow, blood gases or perfusion pressure. AB - This study was performed to determine whether the absorption of liquid from the lungs of postnatal sheep is dependent on pulmonary perfusion pressure, blood gases or blood flow. Relationships between perfusion pressure, rate of lung liquid absorption and perfusate PO2, PCO2 and pH were examined by linear regression analysis from in situ perfused lungs from sheep aged 6 weeks to 6 months. The airspaces of the lungs were filled with liquid containing an impermeant tracer, to allow measurement of the rate of liquid absorption. There was no significant relationship between the rate of lung liquid absorption and pulmonary blood flow (n = 36, r = -0.01, P > 0.1), pulmonary perfusion pressure (n = 36, r = 0.28, P > 0.05) or perfusate PO2, PCO2 or pH. No significant relationships were found between pulmonary blood flow and perfusate PO2, PCO2 or pH. There was no evidence to suggest that the absorption of liquid from the lungs of postnatal sheep is dependent on pulmonary blood flow, blood gases or perfusion pressure, within the limits studied, indicating that lung liquid absorption is dependent on the pulmonary epithelium and not on the pulmonary vasculature. The findings that lung liquid absorption continues in hypoxic environments and despite severe reductions in blood flow may be relevant to the field of transplant surgery. PMID- 11429658 TI - The role of cholinergic and peptidergic pathways in the regulation of pancreatic exocrine function during postnatal development in pigs. AB - The aim of this study was to investigate the role of the parasympathetic (cholinergic and peptidergic) nervous system in the regulation of exocrine pancreas function in piglets during their early postnatal development. The cholinergic and peptidergic regulatory pathways of exocrine pancreatic function were tested by the specific muscarinic receptor blocker 4-diphenylacetoxy-N methylpiperidine-methiodide (4-DAMP) and bombesin, respectively. At the age of 2 weeks, piglets were surgically fitted with a chronic pancreatic duct catheter, a duodenal re-entrant cannula and a jugular vein catheter. The experiments comprised a pre-weaning period, and a post-weaning period that commenced at the beginning of the 5th week of age. Intravenous infusion of 4-DAMP (100 pmol x kg( 1) x h(-1)) reduced the outflow of pancreatic juice, the output of total protein and the activity of trypsin, chymotrypsin, carboxyl ester hydrolase and amylase during preprandial and postprandial pancreatic secretion, in both the pre- and post-weaning periods. However, the inhibitory effect of 4-DAMP during postprandial secretion was significantly greater (P < 0.05) in suckling piglets. The infusion of bombesin (10, 100 and 1000 pmol x kg(-1) x h(-1)) stimulated exocrine pancreatic secretion in a dose-dependent manner during both the pre- and post-weaning periods. However, the stimulatory effect of 1000 pmol x kg(-1) x h( 1) bombesin on total protein output and the activities of trypsin, chymotrypsin and amylase were significantly higher (P < 0.05) in suckling piglets. In summary, our study showed that cholinergic and peptidergic mechanisms are involved in the regulation of exocrine pancreas function in piglets in both the pre- and post weaning stages. 4-DAMP had a greater inhibitory effect on exocrine pancreatic secretion in piglets during the pre-weaning period. Thus, these observations suggest that the parasympathetic nervous system plays a dominant role in the functioning of the exocrine pancreas at this time. The action of bombesin suggests that it is a potent secretagogue for the exocrine pancreas in pigs during their postnatal development. PMID- 11429659 TI - Effects of prior exercise and recovery duration on oxygen uptake kinetics during heavy exercise in humans. AB - Prior heavy exercise (above the lactate threshold, LT) reduces the amplitude of the pulmonary oxygen uptake (VO2) slow component during heavy exercise, yet the precise effect of prior heavy exercise on the phase II VO2 response remains to be established. This study was designed to test the hypotheses that (1) prior heavy exercise increases the amplitude of the phase II VO2 response independently of changes in the baseline VO2 value and (2) the effect of prior exercise depends on the amount of external work done during prior exercise, irrespective of the intensity of the prior exercise. Nine subjects performed two 6 min bouts of heavy cycling exercise separated by 6 min baseline pedalling recovery (A), two 6 min heavy exercise bouts separated by 12 min recovery (6 min rest and 6 min baseline pedalling, B), and a bout of moderate exercise (below the LT) in which the same amount of external work was performed as during the prior heavy exercise, followed by 6 min heavy exercise (C). In both tests A and B, prior heavy exercise significantly increased the absolute VO2 amplitude at the end of phase II (by approximately 150 ml x min(-1)), and reduced the amplitude of the VO2 slow component by a similar amount. Following 12 min of recovery (B), baseline VO2, but not blood [lactate], had returned to pre-exercise levels, indicating that these effects occurred independently of changes in baseline VO2. Prior moderate exercise (C) had no effect on either the VO2 or blood [lactate] responses to subsequent heavy exercise. The VO2 response to heavy exercise was therefore dependent on the intensity of prior exercise, and the effects on the amplitudes of the phase II and slow VO2 components persisted for at least 12 min following prior heavy exercise. PMID- 11429660 TI - Cardiovascular response to graded lower body negative pressure in young and elderly man. AB - Lower body negative pressure (LBNP) reduces central venous pressure (CVP) and cardiac output. The elderly are reported to have a limited capacity to increase cardiac output by increasing heart rate (HR), are especially dependent on end diastolic volume to maintain stroke volume and therefore should be especially vulnerable to LBNP. The present study compared the effects of LBNP in the young and old. Stroke volume was assessed non-invasively as stroke distance (SD) by aortovelography. Two groups of healthy male volunteers were studied: eight young (29.7 +/- 2.0 years, mean +/- S.E.M.) and nine old (70.1 +/- 0.9 years). LBNP was applied progressively at 17.5, 35 and 50 mmHg in 20 min steps, with measurements taken during each steady state. There were similar, significant, falls in CVP in both groups. SD fell significantly in both groups from respective control values of 24.8 +/- 1.6 and 16.6 +/- 0.9 cm to 12.5 +/- 1.3 and 8.9 +/- 0.4 cm at a LBNP of 50 mmHg. Although SD in the elderly was significantly lower than in the young, the LBNP-induced changes were not different between groups. Both groups produced similar significant increases in vascular resistance, HR, plasma vasopressin (AVP) and noradrenaline. Mean arterial blood pressure (MBP) and plasma adrenaline did not change significantly. Therefore healthy old men respond to LBNP in a similar manner to the young, although MBP and SD are regulated around different baselines in the two groups. PMID- 11429661 TI - The Canadian digestive diseases foundation (CDDF) - a message from the president. PMID- 11429662 TI - Acalculous biliary pain: motility dysfunction and functional pain. PMID- 11429663 TI - Infliximab for Crohn's disease: more questions than answers. PMID- 11429664 TI - Stability of human gallbladder bile: effect of freezing. AB - In the present study, the stability of the most essential biliary parameters of human gallbladder bile at -18 degrees C was examined over several months. In 12 patients with gallstone disease (10 female, two male; 52.1+/-13.3 years of age), bile was obtained through fine needle puncture of the gallbladder under local anesthetic. The concentrations of total lipids, cholesterol, phospholipids and bile acids, and the cholesterol saturation index and crystal appearance time were determined before and after freezing over a mean period of 4.38+/-2.9 months. Gallbladder bile obtained by fine needle puncture has proved to be of excellent quality. The total lipid concentration was unchanged before (8.30+/-4.16 g/dL) and after freezing (9.16+/-4.54 g/dL, P=0.6027). The biliary cholesterol, phospholipids and bile acid concentrations, and cholesterol saturation index showed no statistically significant differences before and after freezing. A significant difference arises in the context of subdivision of the group to the nucleation time. Before freezing, most patients had a nucleation time between five and eight days, which shortened to between one and four days after thawing (P=0.0100). The authors conclude that, with the exception of the nucleation time, human gallbladder bile can be stored at -18 degrees C for four months with stability of major lipid components. PMID- 11429665 TI - Canadian Association of Gastroenterology Clinical Practice Guidelines: the use of infliximab in Crohn's disease. PMID- 11429666 TI - Infliximab for the treatment of Crohn's disease: review and indications for clinical use in Canada. AB - Crohn's disease is a chronic inflammatory disorder of the gastrointestinal tract. It may affect any portion of the gastrointestinal tract from the mouth to the anus. Symptoms typically include cramping abdominal pain, diarrhea (which may be bloody) and nausea. As the severity of the illness worsens, patients may experience constant abdominal pain, vomiting, weight loss and fever. From the perspective of the patient, disease symptoms significantly impair quality of life, and interfere with their work environment and activities of daily living. Unfortunately, there is no cure for Crohn's disease. Patients experience a chronic, relapsing course characterized by recurrent flares of their disease. Conventional medical treatment of Crohn's disease includes the use of non specific anti-inflammatory drugs (5-aminosalicylic acid agents, prednisone, budesonide), immunosuppressives (6-mercaptopurine, azathioprine, methotrexate) and antibiotics. A variable onset of action, incomplete response rates and a significant risk of adverse effects characterize current therapies. Although surgery is frequently used to treat complications or medically refractory disease, postoperative recurrence is a common problem. Infliximab, a murine chimeric monoclonal antibody directed toward tumour necrosis factor-alpha, is a highly effective treatment of active Crohn's disease. In randomized, placebo controlled clinical trials, 33% of patients treated with infliximab 5 mg/kg achieved remission (Crohn's Disease Activity Index score less than 150), compared with only 4% of those receiving placebo (P<0.001). Additionally, infliximab is the only drug therapy shown to be effective for the treatment of fistulizing Crohn's disease. In studies done to date, infliximab appears to be well tolerated and has a favourable side effect profile. PMID- 11429667 TI - Anti-TNF therapy and malignancy - a critical review. AB - The roles of tumour necrosis factor (TNF) and anti-TNF therapy in malignancy are reviewed, including an overview of baseline risk factors for malignancy in inflammatory diseases and the incidences of malignancies observed in clinical trials of an anti-TNF-a therapy, infliximab. The preclinical data and early clinical experience presented for infliximab do not provide evidence for a causal relationship between TNF-a antagonism and the development of lymphoid or nonlymphoid cancers. PMID- 11429668 TI - Ranitidine bismuth citrate. AB - Recognition of the relationship between Helicobacter pylori infection and the development of gastroduodenal disease has increased greatly in recent years. To avoid complications of H pylori infection, such as the development of recurrent duodenal and gastric ulcers, effective therapies are required for eradication of the infection. This article reviews ranitidine bismuth citrate (RBC), a novel complex of ranitidine, bismuth and citrate, which was developed specifically for the purpose of eradicating H pylori. Dual therapy with RBC in combination with clarithromycin for 14 days yields eradication rates of 76%. Triple therapy bid for one week with a proton pump inhibitor, clarithromycin and either amoxicillin or a nitroimidazole (tinidazole or metronidazole) is advocated as the treatment of choice for H pylori eradication. Analogous regimens with RBC in place of proton pump inhibitors show effective eradication rates in comparative studies and with pooled data. RBC, used alone or in combination with other antibiotics, appears to be a safe and effective drug for the treatment of H pylori infection. Bismuth levels do not appear to rise to toxic levels. PMID- 11429669 TI - Muscoloskeletal manifestations in inflammatory bowel disease. AB - Muscoloskeletal manifestations are the most common extraintestinal complications of inflammatory bowel disease. Wide ranges in prevalence have been reported, depending on the criteria used to define spondylarthropathy. In 1991, the European Spondylarthropathy Study Group developed classification criteria that included previously neglected cases of undifferentiated spondylarthropathies, which had been ignored in most of the oldest epidemiological studies on inflammatory bowel disease. The spectrum of muscoloskeletal manifestations in inflammatory bowel disease patients includes all of the clinical features of spondylarthropathies: peripheral arthritis, inflammatory spinal pain, dactylitis, enthesitis (Achilles tendinitis and plantar fasciitis), buttock pain and anterior chest wall pain. Radiological evidence of sacroiliitis is common but not obligatory. The articular manifestations begin either concomitantly or subsequent to the bowel disease; however, the onset of spinal disease often precedes the diagnosis of inflammatory bowel disease. The prevalence of the different muscoloskeletal manifestations is similar in ulcerative colitis and Crohn's disease. Symptoms usually disappear after proctocolectomy. The pathogenetic mechanisms that produce the muscoloskeletal manifestations in inflammatory bowel disease are unclear. Several arguments favour an important role of the intestinal mucosa in the development of spondylarthropathy. The natural history is characterized by periods of flares and remission; therefore, the efficacy of treatment is difficult to establish. Most patients respond to rest, physical therapy and nonsteroidal anti-inflammatory drugs, but these drugs may activate bowel disease. Sulphasalazine may be recommended in some patients. There is no indication for the systemic use of steroids. PMID- 11429670 TI - Intestinal carcinoid tumours in a father and daughter. AB - Familial cases of carcinoid tumours that are not associated with any known syndrome or disease are extremely rare. All cases reported in the world literature have involved carcinoid tumours of the gastrointestinal tract. Two cases of carcinoid tumours of the small intestine in a father and daughter are presented. Laboratory analyses did not support the hypothesis that the occurrence of carcinoid tumours in this family is a variant of the multiple endocrine neoplasia type 1 syndrome. A review of the literature on familial occurrence of intestinal carcinoid tumours in the absence of any other known carcinoid tumour predisposing genetic syndrome is provided. PMID- 11429671 TI - Developmental landmarks in offspring of rats exposed singly and in combination to Aroclor 1016 and levothyroxine. PMID- 11429672 TI - Fluoride levels in publicly supplied and bottled drinking water in the Island of Tenerife, Spain. PMID- 11429673 TI - Exposure to endotoxins and microbes in the treatment of waste water and in the industrial debarking of wood. PMID- 11429674 TI - Child-like mannequin for observation and measurement of spray drift particle deposition for pesticide exposure assessment. PMID- 11429675 TI - Correlation of reversed-phase retention factors for alpha-branched phenylsulfonyl acetates with octanol/water partition coefficients and quantum chemical descriptors. PMID- 11429676 TI - Treatment of wastewater from table olive industries: quantum yield of photolytic processes. PMID- 11429677 TI - Tadpole epithelium test: potential use of Rana catesbeiana histopathologic epithelial changes to evaluate aquatic pollution. PMID- 11429678 TI - Biomarker response in Sheim (Acanthopagrus latus) exposed to polycyclic aromatic hydrocarbons. PMID- 11429679 TI - Evaluating the genotoxicity of surface water of Yangzhong City using the Vicia faba micronucleus test and the comet assay. PMID- 11429680 TI - Pesticide residues in selected vegetables and fruits in Alexandria City, Egypt, 1997-1998. PMID- 11429681 TI - Microalgal bioassays as a test of pesticide photodegradation efficiency in water. PMID- 11429682 TI - Gonad organochlorine concentrations and plasma steroid levels in White sturgeon (Acipenser transmontanus) from the Columbia River, USA. PMID- 11429683 TI - Chromium-induced physiologic changes in Vallisneria spiralis L. and its role in phytoremediation of tannery effluent. PMID- 11429684 TI - Copper-influenced changes in lactate dehydrogenase and G-6-PDH activities of the freshwater teleost, Labeo rohita. PMID- 11429685 TI - Effects of zinc on the growth of Bulrush (Schoenoplectus californicus) and its distribution between different sediment fractions. PMID- 11429686 TI - Trace metals in the atmosphere of Niteroi City, RJ., Brazil. PMID- 11429687 TI - Survey of lead in feathers of anatidaes from the Pabellon inlet, Sinaloa, Mexico. PMID- 11429688 TI - Metal concentrations in bivalves living in and around copper mine tailings released after a tailings dam breach. PMID- 11429689 TI - Heavy metals in marine algae from Sile in the Black Sea, 1994-1997. PMID- 11429690 TI - Heavy metal pollution in bottom sediment, Dubai, United Arab Emirates. PMID- 11429691 TI - Copper, chromium, and arsenic levels in soil near highway traffic sound barriers built using CCA pressure-treated wood. PMID- 11429692 TI - Cooperation of Cdc42 small G protein-activating and actin filament-binding activities of frabin in microspike formation. AB - Frabin is a GDP/GTP exchange protein for Cdc42 with actin filament (F-actin) binding activity. Cdc42 is a small GTP-binding protein that forms filopodia-like microspikes in a variety of cells. Expression of frabin indeed forms microspikes through at least activation of Cdc42 in MDCK cells and fibroblasts such as COS7, L, and NIH3T3 cells. However, the role of the F-actin-binding activity of frabin in the microspike formation remains unknown. We have examined here this role of frabin by expressing various frabin mutants, which have lost Cdc42-activating or F-actin-binding activity, with or without a dominant active mutant of Cdc42 in MDCK and COS7 cells. We show here that for the microspike formation, either of the Cdc42-activating and F- actin-binding activities of frabin alone is not sufficient and both the activities are necessary and that both the activities play a cooperative role in the microspike formation. The present results, together with the earlier finding that Cdc42 reorganizes the actin cytoskeleton at least through the N-WASP-Arp2/3 complex, suggest that frabin directly and indirectly reorganizes the actin cytoskeleton through its F-actin-binding and Cdc42-activating activities, respectively, in a cooperative manner, eventually leading to microspike formation. PMID- 11429693 TI - The YXXQ motif in gp 130 is crucial for STAT3 phosphorylation at Ser727 through an H7-sensitive kinase pathway. AB - The signal transducer and activator of transcription (STAT) 3 is essential for mediating signals from the receptors for a variety of cytokines and growth factors, including IL-6 and EGF, and from cytoplasmic tyrosine kinases. Upon stimulation, STAT3 is phosphorylated at Ser727 and Tyr705. However, the role of phosphorylation at Ser727, and the kinase pathways responsible for this phosphorylation in IL-6 signaling remain obscure. Here we show that IL-6 activates at least two distinct STAT3 serine kinase pathways and that an H7 sensitive pathway is dominant over a PD98059-sensitive one in HepG2 cells stimulated with a low concentration of IL-6. The analysis, using a series of chimeric receptors containing the extracellular domain of the G-CSF receptor, the truncated form of gp 130, and additional short peptides at the gp 130 carboxy terminus, showed that the YXXQ motif of gp 130 was sufficient for the H7 sensitive STAT3 Ser727 phosphorylation. This YXXQ-mediated pathway does not involve Erk, p38, JNK, or PKCdelta, and requires a site in the region from 533 to 711 of STAT3 for phosphorylation in vivo. Moreover, we show that Ser727 is required for full transcriptional activity of STAT3 for two different response elements. Thus, the YXXQ motif regulates STAT3 activities in two ways in response to even a low concentration of IL-6: it recruits STAT3 to the receptor for tyrosine phosphorylation, and activates an unidentified H7-sensitive pathway leading to the serine phosphorylation of STAT3. PMID- 11429694 TI - Key role of Shc signaling in the transforming pathway triggered by Ret/ptc2 oncoprotein. AB - The RET/PTC oncogenes, generated by chromosomal rearrangements in papillary thyroid carcinomas, are constitutively activated versions of protoRET, a gene encoding two protein isoforms of a transmembrane tyrosine kinase receptor. By using Ret/ptc2 short isoform (iso9), we have previously demonstrated that Tyr586 (Tyr1062 of protoRet) is the docking site for both the PTB and the SH2 domains of Shc. To determine the relevance of this interaction for the transforming activity of Ret/ptc oncogenes, we have generated and characterized novel Ret/ptc mutants unable to activate Shc: Ret/ptc2 long isoform (iso51)-Y586F and both isoforms of Ret/ptc2-N583A. These mutants neither activate Shc nor transform NIH3T3 cells. Since Tyr1062 shows features of a multifunctional docking site, we have used a Shc mutant (Shc Y317F) to directly assess Shc role. We have demonstrated that in our cell system Shc Y317F behaves like a dominant interfering mutant on the activation of the Grb2-Sos pathway by endogenous Shc triggered by Ret/ptc2. A strong reduction of the transforming activity of Ret/ptc2 in presence of this mutant was also demonstrated. Our data suggest that Shc activation play a key role in the transforming pathways triggered by Ret/ptc oncoproteins. Moreover, we have shown that coexpression of the Shc-Y317F mutant with Ret/ptc2 specifically causes apoptosis, and that the surviving cells lose the long-term expression of one of the two genes. PMID- 11429695 TI - Detection of repair activity during the DNA damage-induced G2 delay in human cancer cells. AB - All eukaryotic cells manifest cell cycle delay after exposure to DNA damaging agents. It has been proposed that such cell cycle checkpoints may allow DNA repair but direct evidence of such activity during the radiation-induced G2 delay has been lacking. We report here that cells arrested in G2 by radiation (2-3 Gy) and etoposide incorporate bromodeoxyuridine (BrdU) at discrete foci in the nucleus. We detected G2 cells with CENP-F, a nuclear protein maximally expressed in G2. Caffeine and okadaic acid, both established radiosensitizers, inhibit the incorporation of BrdU in G2 cells. Radioresistant HT29 and OVCAR cells demonstrate BrdU foci formation more frequently during the G2 delay when compared to the more radiosensitive A2780 cell line. The repair foci formed during G2 may be followed through mitosis and observed in daughter cells in G1. Taken together, these observations are consistent with the detection of DNA repair activity during the radiation-induced G2 delay after relatively low doses of radiation. PMID- 11429696 TI - Downregulation of the potential suppressor gene IGFBP-rP1 in human breast cancer is associated with inactivation of the retinoblastoma protein, cyclin E overexpression and increased proliferation in estrogen receptor negative tumors. AB - The complex insulin-like growth factor network of ligands, receptors and binding proteins has been shown to be disturbed in breast cancer. In addition to defects in proteins controlling cell cycle checkpoints, this type of aberrations could affect tumor growth and survival thereby influencing both tumor aggressiveness and potential response to treatments. We have previously identified the T1A12/mac25 protein, which is identical to the IGFBP-rP1, as a differentially expressed gene product in breast cancer cells compared with normal cells. Here we compare the expression of IGFBP-rP1 in 106 tumor samples with known status of cell cycle aberrations and other clinicopathological data. This was done using a tumor tissue section array system that allows for simultaneous immunohistochemical staining of all samples in parallel. Cytoplasmic staining of variable intensity was observed in most tumors, 15% lacked IGFBP-rP1 staining completely, 20% had weak staining, 32% intermediate and 33% showed strong staining. Low IGFBP-rP1 was associated with high cyclin E protein content, retinoblastoma protein (pRb) inactivation, low bcl-2 protein, poorly differentiated tumors and higher stage. There was a significantly impaired prognosis for patients with low IGFBP-rP1 protein tumors. Interestingly, IGFBP rP1 showed an inverse association with proliferation (Ki-67%) in estrogen receptor negative tumors as well as in cyclin E high tumors suggesting a separate cell cycle regulatory function for IGFBP-rP1 independent of interaction with the estrogen receptor or the pRb pathway. PMID- 11429697 TI - Ectopic expression of cyclin D1 amplifies a retinoic acid-induced mitochondrial death pathway in breast cancer cells. AB - All-trans retinoic acid inhibits growth associated with downregulation of cyclin D1 and can cause low level apoptosis in estrogen receptor positive breast cancer cell lines. The cyclin D1 gene is amplified and/or the protein overexpressed in about one-third of breast cancers. Constitutive expression of cyclin D1 in estrogen receptor positive MCF-7 and ZR-75 breast cancer cells (MCF-7(cycD1) and ZR-75(cycD1)) Increased the fraction of cells in S phase and reduced the G1 accumulation following retinoic acid treatment compared with control cells. However, culture of MCF-7(cycD1) with 1 microM all-trans retinoic acid resulted in about threefold greater growth inhibition compared with vector-transfected cells. Hoechst staining of DNA and in situ DNA end-labeling analysis indicated that MCF-7(cycD1) and ZR-75(cycD1) cultures contained 4-6-fold more retinoic acid induced apoptotic nuclei as vector-transfected cells. Retinoic acid treatment of vector-transfected clones resulted in Bax protein activation as assessed by exposure of the NH(2)-terminus of Bax but the proportion of cells containing activated Bax was increased in cyclin D-expressing cells treated with retinoic acid. The latter cells also displayed both immunocytochemical and biochemical evidence of translocation of cytochrome c into the cytosol following RA treatment. Retinoic acid markedly decreased the Bcl-2 levels in MCF-7 and ZR-75 cells. Accordingly, coexpression of Bcl-2 and cyclin D1 rendered the cells resistant to retinoic acid-induced apoptosis. We conclude that constitutive expression of cyclin D1 sensitizes ER-positive breast cancer cells to a retinoic acid-induced mitochondrial death pathway involving Bax activation, cytochrome c release and caspase-9 cleavage. PMID- 11429698 TI - A sequence element of p53 that determines its susceptibility to viral oncoprotein targeted degradation. AB - The molecular basis that the viral oncoproteins, including HPV16 E6 and E1B55k/E4 34k complex, differentially target p53 but not its homolog p73 for degradation remains elusive. Using a series of p53/p73 chimeras, we demonstrated that despite binding to the different regions of p53, both HPV16 E6 and E1B55k/E4 34k required a very same p53 sequence, amino acid residues 92 to 112 [p53(aa.92-112)], previously identified as a necessity for Mdm2-mediated degradation, to target p53 for degradation. Removal of the p53(aa.92-112) by either substitution or deletion resulted in a p53 protein that was no longer degradable by the viral proteins. More significantly, swapping the oncoprotein-binding motif and the p53(aa.92-112) rendered p73 susceptible to oncoprotein-mediated degradation. Collectively, our data supports a model in which the p53(aa.92-112) functions as a determinant for p53 stability while the binding of the oncoproteins directs p53 into the specific pathway for proteolysis. PMID- 11429699 TI - Adenomatous polyposis coli gene promoter hypermethylation in non-small cell lung cancer is associated with survival. AB - Methylation of 5' CpG islands in promoter and upstream coding regions has been identified as a mechanism for transcriptional inactivation of tumor suppressor genes. The purpose of this study was to determine whether hypermethylation of the adenomatous polyposis coli (APC) gene promoter occurs in primary non-small cell lung cancer (NSCLC), and whether hypermethylated APC has any relationship with survival. APC promoter 1A methylation was determined in normal and corresponding tumor tissue from 91 NSCLC patients and in a control group of 10 patients without cancer, using a quantitative fluorogenic real-time PCR (Taqman) system. APC promoter methylation was detectable in 86 (95%) of 91 tumor samples, but also in 80 (88%) of 91 normal samples of NSCLC patients, and in only two (20%) of 10 normal lung tissues of the control group. The median level of APC promoter methylation was 4.75 in tumor compared to 1.57 in normal lung tissue (P<0.001). Patients with low methylation status showed significantly longer survival than did patients with high methylation status (P=0.041). In a multivariate analysis of prognostic factors, APC methylation was a significant independent prognostic factor (P=0.044), as were pT (P=0.050) and pN (P<0.001) classifications. This investigation shows that APC gene promoter methylation occurs in the majority of primary NSCLCs. High APC promoter methylation is significantly associated with inferior survival, showing promise as a biomarker of biologically aggressive disease in NSCLC. PMID- 11429700 TI - Amifostine (WR2721) restores transcriptional activity of specific p53 mutant proteins in a yeast functional assay. AB - Many p53 mutants found in human cancer have an altered ability to bind DNA and transactivate gene expression. Re-expression of functional p53 in cells in which the endogenous TP53 gene is inactivated has been demonstrated to restore a non tumorigenic phenotype. Pharmacological modulation of p53 mutant conformation may therefore represent a mechanism to reactivate p53 function and consequently improve response to radio- and chemotherapy. We have recently reported that the radio- and chemoprotector Amifostine (WR2721, Ethyol) activates wild-type p53 in cultured mammalian cells. In the present study, we have used a yeast functional assay to investigate the effect of WR2721 on the transcriptional activity of p53. WR2721 restored this activity in a temperature-sensitive mutant V272M (valine to methionine at codon 272) expressed at the non-permissive temperature and it also partially restored the transcriptional activity of several other conformationally flexible p53 mutants. The results indicate that the yeast functional assay may be used to identify compounds that modulate p53 activity, with potential therapeutic implications. PMID- 11429701 TI - Human fibroblast replicative senescence can occur in the absence of extensive cell division and short telomeres. AB - Ectopic expression of telomerase blocks both telomeric attrition and senescence, suggesting that telomeric attrition is a mitotic counting mechanism that culminates in replicative senescence. By holding human fibroblast cultures confluent for up to 12 weeks at a time, we confirmed previous observations and showed that telomeric attrition requires cell division and also, that senescence occurs at a constant average telomere length, not at a constant time point. However, on resuming cell division, these long-term confluent (LTC) cultures completed 15-25 fewer mean population doublings (MPDs) than the controls prior to senescence. These lost divisions were mainly accounted for by slow cell turnover of the LTC cultures and by permanent cell cycle exit of 94% of the LTC cells, which resulted in many cell divisions being unmeasured by the MPD method. In the LTC cultures, p27(KIP1) accumulated and pRb became under-phosphorylated and under expressed. Also, coincident with permanent cell cycle exit and before 1 MPD was completed, the LTC cultures upregulated the cell cycle inhibitors p21(WAF) and p16(INK4A) but not p14(ARF) and developed other markers of senescence. We then tested the relationship between cell cycle re-entry and the cell cycle-inhibitory proteins following subculture of the LTC cultures. In these cultures, the downregulation of p27(KIP1) and the phosphorylation of pRb preceded the complete resumption of normal proliferation rate, which was accompanied by the down regulation of p16(INK4A). Our results show that most normal human fibroblasts can accumulate p16(INK4A), p21(WAF) and p27(KIP1) and senesce by cell division independent mechanism(s). Furthermore, this form of senescence likely requires p16(INK4A) and perhaps p27(KIP1). PMID- 11429702 TI - Deregulated FGFR3 mutants in multiple myeloma cell lines with t(4;14): comparative analysis of Y373C, K650E and the novel G384D mutations. AB - The t(4;14)(p16.3;q32) chromosomal translocation occurs in approximately 20% of multiple myelomas (MM) and leads to the apparent deregulation of two genes located on 4p16.3: the fibroblast growth factor receptor 3 (FGFR3) and the putative transcription factor WHSC1/MMSET. Interestingly, FGFR3 mutations known to be associated with autosomal dominant human skeletal disorders have also been found in some MM cell lines with t(4;14) but their pathogenetic role in MM is still controversial. Since cell lines may represent useful models for investigating the effects of deregulated FGFR3 mutants in MM, we analysed the expression, activation, signaling pathways and oncogenic potential of three mutants identified so far: the Y373C and K650E in the KMS-11 and OPM-2 cell lines respectively, and the novel G384D mutation here identified in the KMS-18 cell line. All of the cell lines present a heterozygous FGFR3 gene mutation and transcribe the mutated allele; unlike KMS-11 and OPM-2 (which express the IIIc isoform), the KMS-18 cell line expresses prevalently the isoform IIIb. We demonstrated that, under serum-starved conditions, KMS-11 and OPM-2 cells express appreciable levels of phosphorylated FGFR3 mutants indicating a constitutive activation of the Y373C and K650E receptors; the addition of the aFGF ligand further increased the level of receptor phosphorylation. Conversely, the FGFR3 mutant in KMS-18 does not seem to be constitutively activated since it was phosphorylated only in the presence of the ligand. In all three MM cell lines, ligand-stimulated FGFR3 mutants activated the MAP kinase signaling pathway but did not apparently involve either the STAT1 or STAT3 cascades. However, when transfected in 293T cells, G384D, like Y373C and K650E, was capable of activating MAPK, STAT1 and STAT3 under serum-starved condition. Finally, a focus formation assay of NIH3T3 cells transfected with FGFR3-expressing plasmid vectors showed that Y373C and K650E (albeit at different levels) but not G384D or the wild-type receptor, can induce transformed foci. Overall, our results support the idea that FGFR3 mutations are graded in terms of their activation capability, thus suggesting that they may play a critical role in the tumor progression of MM patients with t(4;14). PMID- 11429703 TI - The CpG island of the novel tumor suppressor gene RASSF1A is intensely methylated in primary small cell lung carcinomas. AB - Loss of heterozygosity at 3p21.3 occurs in more than 90% of small cell lung carcinomas (SCLCs). The Ras association domain family 1 (RASSF1) gene cloned from the lung tumor suppressor locus 3p21.3 consists of two major alternative transcripts, RASSF1A and RASSF1C. Epigenetic inactivation of isoform A (RASSF1A) was observed in 40% of primary non-small cell lung carcinomas and in several tumor cell lines. Transfection of RASSF1A suppressed the growth of lung cancer cells in vitro and in nude mice. Here we have analysed the methylation status of the CpG island promoters of RASSF1A and RASSF1C in primary SCLCs. In 22 of 28 SCLCs (=79%) the promoter of RASSF1A was highly methylated at all CpG sites analysed. None of the SCLCs showed evidence for methylation of the CpG island of RASSF1C. The results suggest that hypermethylation of the CpG island promoter of the RASSF1A gene is associated with SCLC pathogenesis. PMID- 11429704 TI - Functional characterization of naturally occurring mutants (P405R and P425L) of p73alpha and p73beta found in neuroblastoma and lung cancer. AB - The novel candidate tumor suppressor p73, a structural and functional homolog of p53, activates various p53 responsive promoters and induces tumor cell apoptosis. Although p73 is infrequently mutated in human cancers, we have previously found two types of p73 mutation with amino acid substitution (P405R and P425L) in primary neuroblastoma and lung cancer. Here we report generations of the p73 mutants with either P405R or P425L substitution and functional analysis of these naturally occurring mutants. Indirect immunofluorescence staining revealed that nuclear accumulation of p73alpha or p73beta was not affected by these mutations. The P425L substitution reduced the ability of p73alpha to transactivate various p53 responsive promoters (p21(Waf1), Mdm2, and Bax). Moreover, this down regulation was correlated with the reduced capability of p73alpha(P425L) to suppress cell growth in p53-deficient SAOS-2 cells. In contrast, p73beta(P425L) was as effective as wild-type p73beta in transactivation and growth inhibition. On the other hand, the P405R substitution had no significant effect on both the transcriptional activity and the growth-suppressive ability of p73alpha or p73beta. These results suggested that, at least, one of the naturally occurring p73 mutants, p73alpha(P425L), was a functionally defective mutant of p73. PMID- 11429705 TI - p53 mutants can often transactivate promoters containing a p21 but not Bax or PIG3 responsive elements. AB - The human p53 protein acts mainly as a stress inducible transcription factor transactivating several genes involved in cell cycle arrest (e.g. p21) or apoptosis (e.g. Bax, PIG3). Roughly half of all human tumours contains p53 missense mutations. Virtually all tumour-derived p53 mutants are unable to activate Bax transcription but some retain the ability to activate p21 transcription. Identification of these mutants may have valuable clinical implications. We have determined the transactivation ability of 77 p53 mutants using reporter yeast strains containing a p53-regulated ADE2 gene whose promoter is regulated by p53 responsive elements derived from the regulatory region of the p21, Bax and PIG3 genes. We also assessed the influence of temperature on transactivation. Our results indicate that a significant proportion of mutants [16/77 (21%); 10/64 (16%) considering only tumour-derived mutants] are transcriptionally active, especially with the p21 promoter. Discriminant mutants preferentially affect less conserved (P<0.04, Fisher's exact test), more rarely mutated (P<0.006, Fisher's exact test) amino acids. Temperature sensitivity is frequently observed, but is more common among discriminant than non-discriminant mutants (P<0.003, Fisher's exact test). Finally, we extended the analysis to a group of mutants isolated in BRCA-associated tumours that surprisingly were indistinguishable from wild type in standard transcription, growth suppression and apoptosis assays in human cells, but showed gain of function in transformation assays. The incidence of transcriptionally active mutations among this group was significantly higher than in the panel of mutants studied previously (P<0.001, Fisher's exact test). Since it is not possible to predict the behaviour of a mutant from first principles, we propose that the yeast assay be used to compile a functional p53 database and fill the gap between the biophysical, pharmacological and clinical fields. PMID- 11429706 TI - Msh-2 suppresses in vivo mutation in a gene dose and lesion dependent manner. AB - Mice deficient for the mismatch repair (MMR) gene Msh2 show accelerated tumourigenesis and a reduced apoptotic response to DNA damage of methylation type. Here we examine the effect of mutation for Msh2 on in vivo mutation frequencies in the intestine as determined by loss of function at the Dolichos biflorus (Dlb-1) locus. Spontaneous mutation frequencies were scored in cohorts of ageing mice either wild type or mutant for Msh2. In mice less than 1 year old, mutation frequencies were only elevated in Msh2 null mice. However, beyond this age heterozygous Msh2 mice showed significantly higher mutation frequencies than controls. These findings implicate a gene dose dependent requirement for Msh2 in mutation suppression and prompted an analysis of young Msh2 mutants following exposure to DNA damage. Following exposure to N-methyl-N'-nitro-N nitrosoguanidine (MNNG), Msh2 deficient mice show a reduced apoptotic response and an increase in mutation frequency. Heterozygotes did not differ from controls. Following exposure to cisplatin, no significant elevation was seen in mutation frequencies, even within homozygotes. This is particularly surprising given the association between cisplatin resistance and MMR deficiency. These findings therefore demonstrate a complex reliance upon functional Msh2 in mutation surveillance. We have identified three separate scenarios. First, where retention of both Msh2 alleles over an extended period of time appears critical to the suppression of spontaneous mutation; second, 3 weeks following exposure to MNNG, where only complete loss of Msh2 results in elevated mutation; and finally following cisplatin exposure, where induced levels of mutation are independent of Msh2 status. PMID- 11429707 TI - Contrasting roles of NF-kappaB and JNK in arsenite-induced p53-independent expression of GADD45alpha. AB - Growth arrest and DNA damage-inducible protein 45alpha (GADD45alpha) is an important cell cycle checkpoint protein that arrests cells at G2/M phase by inhibiting the activity of G2-specific kinase, cyclin B/p34cdc2. We report here that arsenite induces GADD45alpha expression in a p53-independent fashion and that this GADD45alpha induction by arsenite is regulated by NF-kappaB and c-Jun-N terminal kinase (JNK) oppositely. In human bronchial epithelial cells overexpressing a kinase-mutated form of IkappaB kinase beta (IKKbeta-KM), the activation of NF-kappaB was inhibited. However, the G2/M cell cycle arrest and expression of GADD45alpha was substantially enhanced in response to arsenite in these cells. Expression of a dominant-negative mutant of SEK1 that blocks JNK activation decreased arsenite-induced GADD45alpha expression. Analysis of GADD45alpha expression in both wild-type and p53-/- fibroblasts indicated that the induction of GADD45alpha by arsenite was independent of the status of p53 protein. PMID- 11429708 TI - HNPCC mutations in the human DNA mismatch repair gene hMLH1 influence assembly of hMutLalpha and hMLH1-hEXO1 complexes. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is a common inherited form of neoplasia caused by germline mutations in DNA mismatch repair (MMR) genes. MMR proteins have been reported to associate with several proteins, including the human exonuclease 1 (hEXO1). We report here novel HNPCC-hMLH1 mutant proteins (T117M, Q426X and 1813insA) in Danish HNPCC patients. We demonstrate that these mutant HNPCC-hMLH1 proteins are unable to form complexes with hEXO1 and hPMS2 in vivo. The results indicate that mutations found in HNPCC gene carriers disrupt hMLH1-hEXO1 complex formation and hMutLalpha heterodimer assembly essential for MMR activity. PMID- 11429710 TI - Breast and cervical cancer survival: making sense of "league tables". AB - During 1998, the Department of Health proposed to use survival rates of cervical and breast cancer in the 1989/90 incidence cohort as indicators of care. Valid interpretation was of concern within Trent and the Trent Cancer Registry responded by performing additional analyses. Trent Cancer Registry registrations for 1989/90 were re-analysed and the stability of districts' ranks for that cohort was investigated using random simulation techniques. Stability of ranks across more recent cohorts was investigated and attempts made to use all available information. The Department of Health's analyses were confirmed by our re-analysis of the 1989/90 cohort: Rotherham residents appeared to have the "worst" survival for cervical cancer, and Sheffield residents for breast cancer, although not statistically significantly so. Random simulations indicated that ranks based on a single cohort are not stable: for example Sheffield (ranked tenth for 1-y breast cancer survival) was ranked third or better in 6% of randomisations. Ranks were also unstable across cohorts: for example Rotherham 1 y cervical cancer survival was ranked tenth for 1989/90, fifth for 1991/92 and tenth for 1993/94. Analysis of 3-y running averages provided better information than the league table approach. Most districts improved over time, to different degrees, and similar sized gaps remained between the "best" and the "worst" districts. This analysis illustrates the need to be circumspect when interpreting "league tables" based on a single year or cohort analysis. League tables are based on ranks: clearly a large difference in rank may reflect only trivial (ie medically unimportant) differences in actual outcome. Lack of a statistically significant difference in survival between two districts does not mean their survival is equivalent. Even for a common cancer, like breast cancer, rankings were unstable from cohort to cohort. At the Registry we propose to perform these trend analyses routinely in future, adjusting, when possible, for the effects of deprivation and stage at diagnosis. PMID- 11429711 TI - Awareness of risk factors for cancer among British adults. AB - Awareness of risk factors for five common cancers was assessed in a representative sample of 3 693 adults in the UK. Respondents were presented with a risk of 14 factors comprising 10 which have established links with various cancers (older age, many sexual partners, low fibre diet, smoking, a relative with the cancer, low fruit and vegetable diet, taking HRT/the contraceptive pill, being overweight, viruses or infection, and a high fat diet) and four so-called "mythic" causes (food additives, overhead power lines, pollution, stress). Out of 15 well-established associations between risk factors and cancers, the average number correctly identified was 5. Women and those with higher levels of education got more correct. Endorsement of mythic causes was comparatively low (less than 5% for most cancers), but were higher in men and those with less education. These results are somewhat reassuring in relation to the prevalence of erroneous beliefs, but raise considerable concern about public understanding of well established causes. Better health education will be needed to maximise public awareness of cancer prevention. PMID- 11429712 TI - Class inequalities in women's health: combined impact of childhood and adult social class--a study of 630 US women. AB - To assess contributions of childhood and adult social class to class gradients in women's health, the authors used gender-neutral household measures of class position in a retrospective cohort study of 630 women enrolled in Examination II of the Kaiser Permanente Women Twins Study (1989-1990, Oakland, CA). The age adjusted odds of reporting fair or poor health was 2.3 times higher (95% confidence interval (CI)=1.2-4.1), using adult class measures, among women categorized as working class vs non-working class/professional. When stratified by childhood social class, however, the elevated risk of fair/poor health among adult working class compared to non-working class/professional women was evident only among those with a non-working class/professional childhood. Similarly, a working class tendency (based on adult class position) towards elevated levels of low density lipoprotein (LDL) cholesterol (odds ratio (OR)=1.5, 95% CI=0.9-2.7) and post-load glucose (OR=1.8, 95% CI=1.0-3.3) was apparent only among women who were non-working class in childhood. These results indicate that both childhood and adult class position influence class gradients in women's health in the United States. PMID- 11429713 TI - Alcohol consumption by non-institutionalised elderly women: the EPIDOS Study. AB - The prevalence of alcohol use declines with age, but studies suggest that between 2% and 4% of the elderly population have a particularly high alcohol consumption. The objective of this study was to verify or refute this finding and identify clinical or social characteristics associated with alcohol consumption. We measured alcohol consumption by autoquestionnaire in 7575 women, aged 75 or older, recruited at five centers in France. The alcohol consumption was computed taking account of the number of beer, wine or liquor (or spirits) drinks consumed per day. The mean age of the respondents was 80+/-6 y. Forty percent used some alcohol and 2.5% drank more than 30 grams per day. Smoking, good health status, higher socioeconomic status or single marital status were factors whose percentages increased significantly with increasing alcohol use. Despite the advanced age of this population, regular alcohol intake was prevalent but not heavy and abusive consumption drinking. Drinking appears to be associated with some medical or social characteristics and possibly with better health status. PMID- 11429714 TI - Smoking and drinking habits as risk factors for hearing loss in the elderly: epidemiological study of subjects undergoing routine health checks in Aichi, Japan. AB - In order to determine the risk factors for hearing loss in the elderly, a total of 496 subjects with bilateral hearing loss and 2807 age-matched persons without hearing disturbance were recruited from the participants in an automated multiphasic health screening examination, and their lifestyle and medical data were analysed. Current smokers showed a significantly increased risk of hearing loss compared with non-smokers (odds ratio after adjustment for sex, age, and potential confounders=2.10 (1.53-2.89)), while heavy drinkers did not show an increased risk compared to non-drinkers. Our findings might provide some clues for the primary prevention of age-related hearing loss. PMID- 11429715 TI - Glucose tolerance and cognitive impairment in an elderly population. AB - We investigated the associations between abnormal glucose tolerance and cognitive impairment in elderly subjects, taking into account some other known determinants of cognitive function. The study population consisted of community-living northern Finnish subjects aged 70 y or over (n=379, of whom were 141 men). Thirty one percent of the men and women (n=43 for the men and n=75 for the women) scored 23 or less in the Mini Mental State Examination. A low level of basic education and high age were the most powerful predictors of impaired cognition. When adjusted for age, gender, educational level, presence of cardiovascular disease (or hypertension), use of alcohol, number of depressive symptoms and poor vision, abnormal glucose tolerance (including IGT) was also weakly associated with impaired cognitive function among these elderly subjects. PMID- 11429716 TI - A comparison of oral fluid collection devices for use in the surveillance of virus diseases in children. AB - Three oral fluid collection devices (OraSure, Omni-SAL and Oracol) were compared in terms of the quality of oral fluid collected by each device for antibody testing and their acceptability to participants. Participants (143 children aged 3.5-5 y from North Hertfordshire, UK, who had recently received DTaP and MMR vaccination) were randomised to use one of the three types of collection device. Oral fluid was collected by a parent who completed a short questionaire recording information on ease of use and willingness to use the device again. A matching serum sample was collected by a nurse. Oral fluid samples were screened for total IgG and IgM by ELISA and for rubella specific IgG and parvovirus specific IgG by radioimmunoassay. Serum samples were screened for rubella specific IgG and parvovirus B19 specific IgG by ELISA. 87.4% (125) of participants provided a matching oral fluid and serum sample. Of these, 100% (125) and 10.4% (13) had serum IgG specific for rubella and parvovirus B19, respectively. The Oracol device provided oral fluid samples with the highest geometric mean titres of total IgG and IgM and with rubella specific IgG results which correlated most closely with those of matching sera. A higher proportion of parents found the Oracol and OraSure devices easier to use than the Omni-SAL (P<0.001) and the proportion who would not take another test was higher for the Omni-SAL than for the Oracol or Orasure. Oral fluid samples collected by each of the devices gave qualitative results acceptable for surveillance and epidemiological studies of rubella and parvovirus B19. The highest quality oral fluid sample for antibody testing in terms of total IgG and IgM concentration and rubella specific IgG concentration was collected by the Oracol. The acceptability to participants of both the Oracol and OraSure was high. As the cheapest device available, the Oracol is the preferred oral fluid collection device for studies involving children in the UK. PMID- 11429717 TI - Sarcoptes scabiei infestation among children in a displacement camp in Sierra Leone. AB - The prevalence of scabies was investigated among a study population of 125 children between the ages of 1-15 y. Confirmation of scabies was done by clinical examination of each child and by the identification of the mite by microscopically examining the scrapings from the scabies-like lesions. The prevalence was age dependent, with children under five years accounting for 77%, peaking to 86% among the 5 to 9-y-olds, and steadily declining with an increase in age. Lesions were found on almost all parts of the body, but lesions were more commonly located on the fingers, legs, hands, face, stomach, and genitalia. Sarcoptes scabiei var hominis was recovered from 84 (67%) of the 125 skin scrapings examined. The prevalence of scabies is high in children in the displacement camps, suggesting that it may be a serious public health problem not only in these camps, but also in the entire country. This may be due to the fact that certain environmental conditions like civil unrest, overcrowding, poor personal hygiene, poverty, and ignorance, which are conducive to the spread of scabies, is characteristically present among the camp residents. Control programs should be put in place and implemented in an integrated nature, by reducing overcrowding, and by improving health education, personal hygiene, treatment and surveillance among high-risk populations. PMID- 11429718 TI - Factors associated with the use of breast and cervical cancer screening services among Chinese women in Hong Kong. AB - We conducted a cross-sectional survey using a self-administered questionnaire among attendees of a well women clinic in Hong Kong during June and July 1998. The study aimed to examine the factors associated with the past and future use of screening services among Chinese women in Hong Kong and their perception of service providers. Of the 430 respondents (64% response rate), 87% were aged 31 50 y, 85% married, 93% attained education to upper secondary school level, and 96% were non-smokers. Nearly all respondents (99%) reported having sexual experience and most of them (87%) had such experience with only one partner; 59% reported having a pap-smear test, 28% a mammogram and 44% a breast self examination. Women who were health conscious (ate a lower fat diet and performed regular exercise) were more likely to have used the screening service (mammogram and pap-smear test) and performed breast self-examination. Staff manner, privacy and cost were the most common contributing factors for respondent's desire for future use of the screening service. Respondents showed a preference for doctors (70%) over nurses (30%), and females (80%) over males (20%) as their service providers. The findings suggest the need to disseminate appropriate information on screening services among the public to dispel misconceptions about the preference for doctors over nurses and females over males. Improving clinician- and other staff-patient communication would be important for breast and cervical screening programs in Hong Kong. PMID- 11429719 TI - Screening: the legal view. AB - Screening has become central to the effective prevention of several diseases, but implementation suffers from difficulties with targeting and rates of compliance. Such issues are also complicated by the need to consider legal provisions regarding confidentiality of patients and other human rights issues. Screening has been an inexact science in relation to, e.g., faecal occult blood testing for colorectal cancer, false positive and false negative tests for HIV, and there have been inadequate quality controls in breast cancer screening programmes. The public need to be made aware of what the screening programmes really offer, balanced against the expectations they may have. There needs to be a clearer understanding of the nature of the contractual and other legal rights of patients/consumers as against providers. A positive screening test may carry adverse consequences as well as benefits. It could alert an insurance company to a risk and lead to additional weighting or even outright rejection for life insurance policies. Job prospects may also be affected for employees. The method of informing patients in relation to screening and screening failure has already been considered by the courts. Realistic information about both screening and treatment efficiency needs to be offered to patients so that they can have a real understanding of what can and cannot be achieved by current science. The development of understanding of the human genome makes the need for clearer legislation in this are more urgent. PMID- 11429720 TI - Contracting for smoking and pregnancy interventions: current practice across England. AB - In order to understand the commissioning of smoking and pregnancy interventions across England prior to the implementation of the Government's national strategy, "Smoking Kills: A White Paper on Tobacco" in 1998, a postal survey was undertaken amongst all 96 Health Authorities (purchasers) and 207 Maternity Service Provider Units (providers) in England. The main outcome measures included the type and duration of contract agreements/service specifications, the level and nature of smoking and pregnancy interventions, barriers to commissioning smoking and pregnancy interventions, data collection and monitoring of activity and quality. A quarter of health authorities were encouraging smoking cessation through contract agreements. The level and complexity of contract agreements and service specifications varied tremendously. Existing smoking and pregnancy interventions were diverse and ad hoc. Data collection and monitoring were haphazard and inconsistent making cross-country comparisons difficult. The commissioning of smoking and pregnancy interventions across England during 1997 and 1998 appeared to be inadequately prioritised. These findings offer a benchmark for observing changes in practice following the recent change of government policy. PMID- 11429721 TI - Does obesity contribute as much to morbidity as poverty or smoking? AB - The prevalence of obesity is increasing in America, but its impact on morbidity relative to other health risks is unclear. This paper compares the effects of overweight, poverty, smoking and problem drinking on occurrence of chronic conditions and health-related quality of life. The data were collected from a nationally representative household telephone survey of 9585 adults fielded in 1998, using self-reported measures of height and weight, poverty, smoking status, problem drinking, chronic conditions and SF-12 global scales. Regression analyses were used to estimate effects of health risk factors on morbidity. Thirty-six percent of adults are overweight but not obese (25< or =BMI<30) and another 23% are obese (BMI> or =30). Controlling for demographics, obesity is associated with more chronic conditions and worse physical health-related quality of life (P<0.01). Smoking history and poverty predict having chronic conditions, but their effect sizes are significantly smaller. Even after controlling for chronic conditions, obesity predicts physical health-related quality of life, in that case with an effect size similar to poverty. The effect of problem drinking is always smaller. Obesity is highly prevalent and associated with at least as much morbidity as are poverty, smoking and problem drinking. Nevertheless, the latter have achieved more consistent attention in recent decades in clinical practice and public health policy. PMID- 11429722 TI - Factors associated with overweight and central body fat in the city of Rio de Janeiro: results of a two-stage random sampling survey. AB - The purpose of the survey was to investigate the association of overweight (body mass index, BMI) and central body fat distribution (waist/hip girth ratio WHR) with socio-economic, demographic, lifestyle and dietary variables in the adult population of Rio de Janeiro City, Brazil, 1995-1996. A two-stage random sample population-based survey was performed, with 1455 males and 1906 females above 20 y old resident in Rio de Janeiro City, Brazil. Data were obtained by direct interview and physical examination of the subjects. The intake of selected nutrients (fat, saturated fat, cholesterol) and energy was obtained from a semi quantitative food frequency questionnaire (FFQ). Variables with at least a marginal univariate association with the dependent variables (BMI and WHR) were selected as predictors in two logistic regression models, and variables statistically significant (P<0.05) were retained in them. Overweight prevalence was 44.9%, and 39.2% of the subjects had excessive central body fat distribution (elevated WHR). The proportions of subjects with an excessive intake of fat, saturated fat and cholesterol were respectively 31%, 42% and 47%. For the BMI model, the variables retained were age (OR=1.5, 95% CI=1.3-1.7), schooling (OR=1.7, 1.4-1.9) and smoking (OR=0.8; 0.7-0.9); and for WHR, age (OR=1.8, 1.5 2.1), schooling (OR=2.2, 1.9-2.6), occupation (OR=1.8, 1.1-2.0) and gender (OR=3.9, 3.2-4.7). Obesity and excessive central body fat are highly prevalent health problems in the studied population. As suggested by the identified risk factors, they should be urgently addressed through health nutrition education and physical activity programs; particularly those directed to the middle aged and female groups. PMID- 11429723 TI - Treatment after rodent exposure necessary to avoid death from rabies. PMID- 11429725 TI - Preventing drug diversion. PMID- 11429726 TI - The use of enucleation and liquid nitrogen cryotherapy in the management of odontogenic keratocysts. AB - PURPOSE: This study evaluated the use of enucleation and cryosurgery in the management of odontogenic keratocysts. PATIENTS AND METHODS: This study involved a retrospective review of 26 patients. All of the patients received a combination of enucleation and cryosurgery. Postoperative follow-up consisted of clinical and radiographic examinations. RESULTS: Before enucleation and cryotherapy, 22 of the 26 patients had received previous treatment consisting of enucleation alone. The average time from initial treatment to recurrence was 6.2 years. Twenty-three cases occurred in the mandible, 22 in the posterior (proximal to the canine), and 1 in the anterior mandible. Three cases involved the maxilla. Three of the 26 patients (11.5%) developed a recurrence after treatment. The average time from treatment to recurrence in these 3 patients was 1.6 years (range, 1.2 to 1.9 years). The remaining 23 patients (88.5%) had no evidence of clinical or radiographic recurrence. The average time of follow-up was 3.5 years (range, 2.0 to 10.0 years). CONCLUSIONS: Based on these results, the combination of enucleation and liquid nitrogen cryotherapy may offer patients improved therapy in the management of odontogenic keratocysts. PMID- 11429728 TI - Intraoral distraction osteogenesis of the mandible in hemifacial microsomia. AB - PURPOSE: Lengthening of the mandible by distraction osteogenesis is the preferred method for treatment of hemifacial microsomia in children. Use of an intraoral distraction technique and horizontal oblique ramus osteotomy in such patients is presented. PATIENTS AND METHODS: Mandibular ramus lengthening was performed in 11 patients aged 6 to 12 years with hemifacial microsomia. During the age of mixed dentition in hemifacial microsomia patients with a hypoplastic mandible, the unerupted molars buds are located high in the retromolar region and are in danger of being damaged by the osteotomy. Therefore, an intraoral approach exposing the mandibular ramus and angle was performed, and a horizontal oblique ramus osteotomy was made, preserving the inferior alveolar nerve. An intraoral device was placed along the ramus, and distraction was started on the third postoperative day at the rate of 1 mm/d and continued for 2 to 3 weeks or as long as necessary. The device was maintained for retention an additional 6 weeks and was then removed. RESULTS: Clinically, the face became more symmetric. The postdistraction posteroanterior cephalometric radiographs demonstrated elongation of the affected ramus and improvement in facial symmetry. CONCLUSIONS: The advantages of this method are that it allows device placement along the ramus, permitting the ramus elongation necessary in treatment of hemifacial microsomia, that it prevents damage to the tooth buds which, during the age of mixed dentition, are in a higher position in the retromolar area, and that it prevents injury of the inferior alveolar nerve. PMID- 11429730 TI - Clinical evaluation of 3 types of plate osteosynthesis for fixation of condylar neck fractures. AB - PURPOSE: The goal of this study was to evaluate outcomes in patients who had condylar neck fractures treated with 3 different plating techniques. PATIENTS AND METHODS: A retrospective study was performed on 37 patients with 40 fractures of the condylar neck that were reduced and stabilized using an approach involving exposure of the facial nerve. Stabilization was achieved with a single miniplate (17 fractures), a minidynamic compression plate (13 fractures), or double miniplates (10 fractures). RESULTS: Plate fracture or screw loosening was exclusively observed in cases stabilized with either a single miniplate or a minidynamic compression plate. No cases of inadequate stability were observed when 2 miniplates were used. CONCLUSIONS: The 2-miniplate fixation technique provides functionally stable fixation for fractures of the condylar neck. PMID- 11429732 TI - The changing face of odontogenic infections. AB - PURPOSE: The purpose of this investigation was to compare characteristics of patients hospitalized with odontogenic infections during the 1980s to those of the 1990s. PATIENTS AND METHODS: This study was a retrospective record review that compared 2 cohorts of patients admitted to the same institution during two 81-month periods, one decade apart. Admission criteria were face or neck swelling suggesting abscess or cellulitis and one or more of the following: temperature above 38 degrees C, white blood cell (WBC) count greater than 10.8 x 10(3)/microL, or concern about airway compromise. Characteristics reviewed were age, gender, race, admission temperature, admission WBC count, fascial space(s) involved, tooth of etiology, duration of hospitalization, and bacteria isolated. Data were compared for statistical significance (P <.05). RESULTS: No significant differences were found between the 2 cohorts for age, gender, race, admission temperature, admission WBC count, space involvement, or length of stay (P <.05). One tooth (mandibular left first molar) of 52 was involved more frequently in the 1990 group (P <.03). Gram-positive cocci were isolated significantly more frequently from the 1990s patients than from the 1980s patients (P <.03). There were also significant differences (P <.02) between cohorts in the isolation frequency of individual genera, such as alpha-hemolytic Streptococci, coagulase negative staphylococci, Staphylococcus epidermidis, Bacteroides melanogenicus, beta-lactamase positive Bacteroides, Eikenella corrodens, and Neisseria species. Eighty-one percent of the bacteria cultured from the 1990s patients were resistant to one or more common antibiotics; 47% of these organisms were Staphylococcus aureus. CONCLUSION: No clinically significant differences existed in the characteristics of patients hospitalized with odontogenic infections between the 1980s and the 1990s. Although there were differences in the type and prevalence of bacteria isolated, this was probably a result of changes in nomenclature, identification protocols, and isolation techniques. PMID- 11429734 TI - Discectomy as an effective treatment for painful temporomandibular joint internal derangement: a 5-year clinical and radiographic follow-up. AB - PURPOSE: The purpose of this prospective study was to compare the preoperative and 5-year postoperative status of patients after unilateral discectomy for painful internal derangement of the temporomandibular joint. PATIENTS AND METHODS: The study was based on 64 patients. Fifty-six had disc displacement without reduction and 8 had disc displacement with reduction. The patients were examined clinically and radiographically before and 5 years after the operation according to a standardized protocol. RESULTS: Eighty-five percent of the patients (n = 52) had good results, 6% had acceptable results, and 9% had a poor outcome. Three patients (5%) were reoperated and 5 other patients (8%) needed a contralateral operation during the 5-year follow-up. The median increase in maximum mouth opening was 11 mm (range, -8 to +35 mm) and 83% of the patients had more than 40 mm maximal mouth opening postoperatively. Seventy-five percent of the patients had crepitation at 5-year follow-up compared with 27% before the operation. Postoperatively, the majority of the joints showed radiographic evidence of osteophytes, flattening, and sclerosis. The radiographic alterations did not correlate with the patients' symptoms. CONCLUSIONS: The results of this long-term follow-up after unilateral discectomy support the use of this operation in patients with painful internal derangement who show no improvement after prior nonsurgical treatment. The postoperative radiographic alterations should be interpreted as adaptive changes rather than progressive degenerative joint disease. PMID- 11429736 TI - Reconstruction of superficial skin cancer defects of the nose. AB - PURPOSE: This article reviewed the results of reconstruction of surface nasal defects after removal of skin cancer. PATIENTS AND METHODS: One hundred patients with 103 surface defects involving various locations on the nose were treated. Management included direct closure, secondary epithelization, full-thickness skin grafts, and local flaps using 1 or 2 stages. RESULTS: Ten nasal defects were treated by direct closure, 8 defects healed by secondary epithelization, and 30 patients were treated with a full-thickness skin graft. Fifty-five defects were reconstructed with local flaps including 30 one-stage and 25 two-stage flaps. CONCLUSION: Many options are available for reconstructing nasal defects that can lead to acceptable aesthetic results. Among the factors that need to be addressed before choosing a procedure for reconstruction of surface nasal defects resulting from skin cancer are size and location of the defect, aesthetic concerns, and the medical status of the patient. PMID- 11429737 TI - Condylar process fractures in children: a follow-up study of fractures with total dislocation of the condyle from the glenoid fossa. AB - PURPOSE: The purpose of this study was to investigate the long-term clinical and radiologic outcome of nonsurgically treated, dislocated condylar fractures sustained during childhood. PATIENTS AND METHODS: Dislocated condylar process fractures were diagnosed in 34 children aged 15 years or younger from 1980 to 1991. Of these, 26 had been treated nonsurgically and were asked to participate in a follow-up examination. Eighteen patients, representing 69% of the total sample, took part in the study. All patients underwent a clinical investigation with special emphasis on temporomandibular joint (TMJ) function. The patients also underwent a radiologic investigation, focusing on fracture remodeling and symmetry of the mandible. RESULTS: After a follow-up period ranging from 4.8 to 16.4 years (mean, 8.6 years), 56% had some subjective symptoms, and 72% had some objective signs of TMJ dysfunction. In general, however, the symptoms and signs of dysfunction were very slight. No correlation was observed between the method of nonsurgical treatment and the clinical results. Radiologic investigation showed incomplete remodeling (76.5%) and asymmetry of the mandible (64.7%) in most patients. The asymmetry was slight, however, and could not be observed clinically. CONCLUSIONS: Conservative treatment of dislocated condylar process fractures in children results in satisfactory long-term outcome of jaw function despite a high frequency of radiologically noted aberrations. Soft diet with immediate mobilization seems to be the treatment of choice. PMID- 11429739 TI - Experimental validation of a finite element model of the temporomandibular joint. AB - PURPOSE: A 2-dimensional finite model of the temporomandibular joint (TMJ) was previously developed to provide a way of studying the specific roles of individual components as well as the overall dynamics of joint motion. This study was undertaken to show that the previously reported finite element model provides results that are consistent with the experimentally obtained results. MATERIALS AND METHODS: The upper compartment of a TMJ of a fresh cadaver specimen was exposed to allow the insertion of a small strip of pressure-sensitive film. Measured loads were applied to the chin and angle of the mandible, pressing the condyle into the glenoid fossa. The resulting stresses in the joint stained the film, providing a way to determine their magnitude. Similar loads were applied to the finite element model and the stresses in the TMJ were mathematically calculated. RESULTS: Experimental results were successfully obtained in 4 separate attempts, recording maximum stresses of 5.6, 8.6, 6.4, and 9.9 MPa (megapascals), respectively. The corresponding finite element model predictions were 7.3, 6.9, 6.4, and 8.2 MPa, respectively. CONCLUSIONS: This study indicates that the results of the previously reported finite element model of the TMJ provide a reasonable approximation of the actual physical situation. PMID- 11429740 TI - Bending of the distraction site during mandibular distraction osteogenesis in the rabbit: a model for studying segment control and side effects. AB - PURPOSE: The purpose of this investigation was to develop an animal model for studying and correcting mandibular distraction side effects. MATERIALS AND METHODS: Twenty-nine rabbits underwent bilateral mandibular distraction. Bending of the mandible was accomplished by offsetting a linear distraction by 35 degrees from the occlusal plane (4 screws per distractor), rotating the anterior segment inferiorly (2 screws per distractor), and removing a 3- or 6-mm wedge of the distraction site. The amount of bite opening varied according to the surgical design. Direct measurements, radiographs, and histology samples were compared. RESULTS: Linear distraction produced a 4-mm anterior open bite and a Class III malocclusion after 2 weeks of distraction. Segmental rotation produced an 8-mm anterior open bite without complications. Removal of a wedge initiated rotation of the anterior segment. A large wedge (6 mm) produced fibrous union in the distraction site. The amount of bite opening or closure depended on the number of surgical screws and position of the distractor. Serial histologic sections showed bone formation at the rotated, triangular distraction site. CONCLUSION: Bite opening or closure can occur from loss of segment control or by altering surgical design. This information is needed to counter unwanted side effects or to plan segment rotations. PMID- 11429741 TI - Conservative management of the fractured atrophic edentulous mandible. PMID- 11429742 TI - Invasive management of the fractured atrophic edentulous mandible. PMID- 11429743 TI - Multiple slow-growing nodules on the cheek. PMID- 11429744 TI - Cephalic tetanus following minor facial abrasions: report of a case. PMID- 11429745 TI - Schwannoma of the tongue: report of 2 cases. PMID- 11429746 TI - Gingival cancer suspected of producing granulocyte colony-stimulating factor: report of a case. PMID- 11429747 TI - Solitary fracture of the articular eminence: a case report. PMID- 11429748 TI - Peripheral ameloblastoma: review of the literature and report of recurrence as severe dysplasia. PMID- 11429749 TI - Nonossifying fibroma of the mandible in a 6-year-old girl: a case report and review of the literature. PMID- 11429750 TI - Follicular carcinoma of the thyroid gland with initial presentation as an oral soft tissue mass: report of a case. PMID- 11429751 TI - Atypical histiocytic granuloma of the oral mucosa: an unusual clinicopathologic entity simulating malignancy. PMID- 11429752 TI - Cellular schwannoma of the mandible: a case report. PMID- 11429753 TI - Treatment of a giant mandibular arteriovenous malformation with percutaneous embolization using histoacrylic glue: a case report. PMID- 11429754 TI - Pseudomonas parotid abscess. PMID- 11429755 TI - Recurrent osseous choristoma of the masseter muscle: case report. PMID- 11429756 TI - Segmental alveolar distraction: the same or different technique. PMID- 11429758 TI - Freeze tolerance in the gray treefrog: cryoprotectant mobilization and organ dehydration. AB - Freeze tolerance in the frog Rana sylvatica is supported by nonanticipatory mobilization of cryoprotectant (glucose) and redistribution of organ water. Other freeze-tolerant frogs may manifest these responses but differences exist. For example, the gray treefrog (Hyla versicolor) accumulates mostly glycerol as opposed to glucose. The current study reports additional novel features about cryoprotection in H. versicolor. Frogs were acclimated to low temperature for 12 weeks and frozen for 3 days at -2.4 degrees C. Some frogs were then thawed at 3 degrees C for 4 hr. Calorimetry revealed that frozen frogs had 53.9% +/- 11.1% of their body water in ice, and all frogs recovered following this procedure. Plasma glucose was low prior to the onset of freezing (1.1 +/- 0.9 micromol/ml) and it was 20x higher in postfreeze frogs. Constituting nearly 30% of plasma solute, glycerol was 117.2 +/- 13.6 micromol/ml prior to freezing and it remained equally high in postfreeze frogs. Liver water content was moderately lower in frozen frogs when compared to controls (62.9% +/- 3.7% vs. 68.6% +/- 1.7%), whereas postfreeze frogs excessively hydrated their livers (75.7% +/- 2.1%). Less pronounced changes were seen in muscle water content. H. versicolor can mobilize its major cryoprotectant, glycerol, in response to extended cold acclimation, which is unique in comparison to other freeze-tolerant frogs, and it experiences only moderate organ dehydration during freezing. This species conforms with other freeze-tolerant frogs, however, by mobilizing glucose as a direct response to tissue freezing. PMID- 11429759 TI - Nonuniform distribution of myosin light chains within the thick filaments of lobster slow muscle: Immunocytochemical study. AB - The in situ distribution of the alpha and beta myosin light chains was investigated at the subsarcomeric and subfilament levels in individual fibers of the superficial flexor muscle (SFM) of the lobster, Homarus americanus. Polyclonal antibodies were produced against the two classes of myosin light chains and used for subsequent immunolocalization on thin sections of sarcomeres and on isolated filaments from both the medial and lateral fiber bundles of the SFM. The beta myosin light chains were uniformly distributed within the crossbridge region of sarcomeres of both medial and lateral bundles. The alpha myosin light chains were uniformly distributed within the crossbridge region of sarcomeres from the medial bundle, but were nonuniformly distributed over the crossbridge region of lateral bundle sarcomeres. In the latter, the number of alpha myosin light chains was highest toward the center of the thick filaments, diminishing towards the ends. Similar distributions of alpha light chains were found in isolated myosin filaments. These data demonstrate that heterogeneity in protein composition extends to the level of the myosin filament and suggest that the myosin filament substructure in lobster may be different than that found in vertebrate skeletal muscle. PMID- 11429760 TI - Tissue interactions and antlerogenesis: new findings revealed by a xenograft approach. AB - Tissue interactions play a pivotal role in organogenesis. Here we describe a xenograft approach to investigate how heterotypic tissue interactions control antler formation in deer. Deciduous antlers grow from the apices of permanent protuberances, called pedicles. Histogenesis of pedicles depends on the antlerogenic periosteum (AP). Pedicles and growing antlers are made up of interior osseocartilage (a mixture of bone and cartilaginous tissue) and exterior skin. In a previous study we hypothesised that pedicle growth may result from mechanical interactions between the interior and exterior components whereas antler generation from a pedicle would involve molecules communicating between the interior and exterior components. To test this hypothesis, we subcutaneously transplanted AP of red deer (Cervus elaphus), either alone or with future pedicle skin, onto nude mice. The results showed that under the nude mouse skin, subcutaneously xenografted AP alone not only could form pedicle-shaped protuberances but also could differentiate into well-organised pedicle-like structures. The overlying mouse skin accommodated the expansion of the grafted AP by initial mechanical stretching and subsequent formation of new skin. Nude mouse skin was not capable of participating in antler tissue formation. However, grafted deer skin together with AP may have successfully rescued this failure after wounding, which highlights the necessity of the specificity of the overlying skin for antler tissue generation. Therefore, we conclude that it is the interaction between the antlerogenic tissue and the overlying skin that results in antlerogenesis: reciprocal mechanical interactions cause pedicle formation, whereas reciprocal instructive interactions induce first antler generation. PMID- 11429761 TI - Metamorphosis of summer flounder, Paralichthys dentatus: cell proliferation and differentiation of the gastric mucosa and developmental effects of altered thyroidal status. AB - Summer flounder, like most marine fishes studied to date, are stomachless at first feeding, and subsequently acquire gastric function during the process of metamorphosis. Stomach formation is controlled largely by thyroxine (T4). In the present work we sought to understand gastric organogenesis in terms of cell proliferation and its relationship to histological differentiation. The objectives of the study were (1) to obtain a developmental pattern of cell proliferation in the gastric mucosa and to relate that pattern to the progress of gastric differentiation; and (2) to understand the regulatory role of T4 on cell proliferation and histological differentiation by altering the thyroidal status of the developing larvae. We observed that (1) in normally developing larvae, cell proliferation increased by early metamorphic climax (MC), remained high until mid-MC, and decreased to basal levels by late MC; concomitantly, the gastric glands appeared and differentiated in the fundic mucosa, and were complete by late MC; (2) T4 accelerated the differentiation of gastric glands and mucus neck cells, while inhibiting the concomitant increase in cell proliferation observed in controls; and (3) the goitrogen thiourea inhibited both cell proliferation and gastric differentiation compared to controls. These results indicate that T(4) is necessary for the three-fold increase in cell proliferation that occurs in early metamorphic climax, but that high T4 levels promote differentiation at the expense of proliferation. The observed effects would be consistent with the normal, metamorphosis-related increase in whole body T4. PMID- 11429762 TI - Effects of feeding high dosages of vitamin E to laying hens on thyroid hormone concentrations of hatching chicks. AB - Long-term experimental feeding of 20,000 ppm alpha-tocopheryl acetate to laying hens caused a significant (P < 0.05) decrease in hatching rates as compared to the control group, which was fed a diet containing 19 ppm alpha-tocopherol. When the thyroid hormones in the developing chicks were checked on incubation days 16, 19, 21, and 22, the following results were ascertained: During the latter part of incubation, increases in plasma concentrations of thyroxine and triiodothyronine were observed. No significant differences in hormone concentrations (P > 0.05) between the control and the treatment group were observed during incubation days 16, 19, and 22. However, on the day of hatching (day 21 of incubation) significantly lower (P < 0.05) triiodothyronine concentrations in chick embryos of piped eggs were found in the treatment group. Moreover, thyroxine concentrations in non-piped eggs and in hatched chicks were found to be significantly higher as compared to the control group. Given these results, one concludes that extremely high dosages of vitamin E may affect thyroid hormone concentrations of hatching chicks, and therefore, the chicks might be inhibited in pipping the egg shell. Hypothetically, the hepatic enzyme 5'-monodeiodinase is involved in the mechanism of inhibition. PMID- 11429763 TI - Embryonic and postembryonic neurogenesis in the ventral nerve cord of the freshwater crayfish Cherax destructor. AB - Previous studies of neurogenic activity in the thoracic neuromeres of indirect developing crustaceans indicated that the temporal patterns of neurogenesis can be correlated with the appearance of the thoracic appendages during larval and metamorphic development. To test further the idea that the temporal patterns of neurogenesis in crustaceans are related to their life histories, we examined neurogenesis in the ventral nerve cord of a direct developing crustacean, the freshwater crayfish Cherax destructor, whose life history contains neither larval stages nor metamorphoses. Neurogenesis was examined using the in vivo incorporation of bromodeoxyuridine into DNA. During late embryonic development the thoracic neuromeres of the crayfish contain arrays of mitotically active neuroblasts similar to those previously described in the spider crab and lobster. The arrays in the crayfish abdomen are, however, greatly reduced compared with those of the thorax. On hatching, both the thoracic and abdominal appendages of C. destructor are capable of movement. The pleopods, however, do not beat rhythmically until the second postembryonic stage whereas the pereiopods are not used in coordinated walking movements until the third stage. An examination of the time course of neurogenesis in the ventral nerve cord revealed that neurogenic activity in each neuromere ceases during or before the moult to the developmental stage in which its segmental appendage is first used in coordinated movements. These findings indicate that the patterns of neurogenesis in crustaceans are indeed related to the maturation of the segmental appendages and, in particular, to the maturation of motor behaviours. PMID- 11429764 TI - Compaction in preimplantation mouse embryos is regulated by a cytoplasmic regulatory factor that alters between 1- and 2-cell stages in a concentration dependent manner. AB - Present studies were performed to investigate what factors affect the morphogenesis of preimplantation mouse embryos, and to find the action mechanism of that factor by using cytoplasm removal and its reconstitution from a different developmental stage embryo. Half (HP group) or one-third of cytoplasm (TP group) was removed from 1-cell mouse embryos by micromanipulation, and their morphogenesis and genome expression were compared with sham-operated embryos (SP group). The compaction and blastocoel formation of embryos in both the HP and TP groups were accelerated in time and cell stage when compared with those of the SP group. However, the total activity and time of RNA synthesis, and gene expression of ZO-1alpha+ isoform were not different. To change the cytoplasm composition without altering the nucleus/cytoplasmic ratio, half a 1-cell embryo with both pronuclei was reconstituted with the half enucleated cytoplasm of 1-cell embryo (P + P group), 2-cell (P + 2 group) or 4-cell (P + 4 group) by electrofusion. Embryonic compaction, timing of RNA synthesis, and stage-specific gene expression of the ZO-1alpha(+) isoform in the P + 2 and P + 4 groups were accelerated in time and cell stage than that in the P + P group, but not different between the P + 2 and P + 4 groups. In addition, a blastomere of 2-cell embryo was reconstituted with the enucleated cytoplasm of 1-cell embryo (2 + P group) or 2 cell (2 + 2 group) in equal volume by electrofusion. Also, the karyoplast of 2 cell was fused with the enucleated 1-cell embryo (2 + PP group). Embryonic development, total activity of RNA synthesis, and gene expression of the ZO 1alpha(+) isoform of embryos in the 2 + P and 2 + PP groups were delayed when compared with those of the 2 + 2 group. Also, the phenomena of compaction and blastocoel formation were delayed in the development time and cell stage. From these results, the nucleus/cytoplasm ratio was found to have no direct effect on the regulation of embryonic morphogenesis, although it accelerated compaction and blastocoel formation. However, cytoplasmic factors that altered between 1- and 2 cell stages regulate embryonic morphogenesis, especially compaction, of preimplantation mouse embryos in concentration-dependent manner. PMID- 11429765 TI - Flow cytometry in the marine environment. Proceedings from the 20th ISAC Congress. Montpellier, France, 20-25 May 2000. PMID- 11429766 TI - Flow cytometry in oceanography 1989--1999: environmental challenges and research trends. AB - BACKGROUND: The present review is based on the identification of four major environmental crises that have been approached from a biological oceanographic viewpoint. These crises are the release of contaminants in near shore marine waters, the collapse of marine resources that were renewable until recently, the loss of biodiversity, and global climate change METHODS: The review examines the contribution of cytometry-based biological oceanography to the resolution of the four environmental crises. Using a database of 302 papers, flow cytometric (FCM) studies in biological oceanography over the 1989--1999 decade are examined. Future biological oceanographic applications of FCM are discussed. RESULTS: Most of the published FCM oceanographic studies focus on phytoplankton and bacterioplankton. Analysis of our 1989-1999 database shows the predominance of studies dedicated to phytoplankton (77%), followed by heterotrophic bacteria (21%). The latter progressively increased over the last decade, together with the improved understanding of the biogeochemical and trophic roles of marine bacteria. Most studies on these two microorganisms were conducted in vitro until 1996, after which the trend reversed in favor of in situ research. The most investigated areas were those with major international sampling efforts, related to the changing climate. Concerning environmental topics, 62% of papers on phytoplankton and bacterioplankton focused on the structure of microbial communities and fluxes (e.g., production, grazing); this provides the basis for biological oceanographic studies on resources and climate change. CONCLUSIONS: Future progress in the biological oceanographic use of FCM will likely fall into two categories, i.e., applications where FCM will be combined with the development of other methods and those where FCM will be the main analytical tool. It is expected that FCM and other cytometric approaches will improve the ability of biological oceanography to address the major environmental challenges that are confronting human societies. PMID- 11429767 TI - Effects of mismatched refractive indices in aquatic flow cytometry. AB - BACKGROUND: Forward-angle light scatter, as measured by flow cytometry, can be used to estimate the size spectra of cell assemblages from natural waters. The refractive index of water samples from aquatic environments can differ because of a variety of factors such as dissolved organic content, aldehyde preservative, sample salinity, and temperature. In flow cytometric analyses, mismatch between the refractive indices of the sheath fluid and the sample causes distortion of the forward-angle light scatter signal. We measured the effect of this mismatch on cell size measurements. METHODS: We examined the error by measuring the scatter signal of a variety of particle types and sizes and changing the sheath to-sample salinity ratio. The effects were characterized for standard microspheres, cultured phytoplankton cells of different sizes, and natural populations from an estuarine river. RESULTS: We found that the distorted scatter signals resulted in an increase in the apparent size of small cells (1--2 microm) by a factor of 4.5 times. Cells in the size range of 3--5 microm were less affected by the salinity differences, and cells larger than 5 microm were not affected. Chlorophyll and phycoerythrin fluorescences and 90 degrees light scatter signals were not changed by sheath and sample salinity differences. CONCLUSIONS: Care must be taken to ensure that the sheath and sample refractive index are matched when using forward light scatter to measure cell size spectra, especially in estuarine studies, where salinity can vary greatly. Of the factors considered that can change the sample refractive index, salinity gradients in an estuary cause the largest index mismatch and, consequently, the largest error in scatter. PMID- 11429768 TI - An industrial application of multiparameter flow cytometry: assessment of cell physiological state and its application to the study of microbial fermentations. AB - BACKGROUND: When using traditional microbiological techniques to monitor cell proliferation and viability, stressed, sublethally injured, or otherwise "viable but nonculturable" cells often go undetected. Because of this, such cells often are not considered by mathematical models used to predict bioprocess performance on scale-up and inaccuracies result. Therefore, analytical techniques, decoupled from postsampling growth, are desirable to rapidly monitor individual cell physiologic states during microbial fermentations. METHODS: Microbial cells, including Escherichia coli, Rhodococus sp., and Sacharomyces cerevisiae, were taken at various stages from a range of fermentation processes and stained with one of three mixtures of fluorescent stains: rhodamine 123/propidium iodide, bis oxonol/propidium iodide, or bis-oxonol/ethidium bromide/propidium iodide. An individual cell's physiologic state was assessed with a Coulter Epics Elite analyzer based on the differential uptakes of these fluorescent stains. RESULTS: It was possible to resolve an individual cell's physiologic state beyond culturability based on the functionality of dye extrusion pumps and the presence or absence of an intact polarized cytoplasmic membrane, enabling assessment of population heterogeneity. This approach allows the simultaneous differentiation of at least four functional subpopulations in microbial populations. CONCLUSIONS: Fluorescent staining methods used in our laboratories have led to a functional classification of the physiological state of individual microbial cells based on reproductive activity, metabolic activity, and membrane integrity. We have used these techniques extensively for monitoring the stress responses of microorganisms in such diverse areas as bioremediation, biotransformation, food processing, and microbial fermentation; microbial fermentation is discussed in this article. PMID- 11429769 TI - Using light scatter signal to estimate bacterial biovolume by flow cytometry. AB - BACKGROUND: In the past decade, flow cytometry has become a useful and precise alternative to microscopic bacterial cell counts in aquatic samples. However, little evidence of its usefulness for the evaluation of bacterial biovolumes has emerged in from the literature. METHODS: The light scattering and cell volume of starved bacterial strains and natural bacterial communities from the Black Sea were measured by flow cytometry and epifluorescence microscopy, respectively, in order to establish a relationship between light scattering and cell volume. RESULTS: With the arc-lamp flow cytometer, forward angle light scatter (FALS) was related to cell size in both the starved strains and natural communities, although regression parameters differed. We tested the predictive capacity of the FALS verous cell size relationship in a bacterial community from the North Sea. That analysis showed that a reliable bacterial biovolume prediction of a natural bacterial community can be obtained from FALS using a model generated from natural bacterial community data. CONCLUSIONS: Bacterial biovolume is likely to be related to FALS measurements. It is possible to establish a generally applicable model derived from natural bacterial assemblages for flow cytometric estimation of bacterial biovolumes by light scatter. PMID- 11429770 TI - Pattern recognition in flow cytometry. AB - BACKGROUND: Analytical flow cytometry (AFC), by quantifying sometimes more than 10 optical parameters on cells at rates of approximately 10(3) cells/s, rapidly generates vast quantities of multidimensional data, which provides a considerable challenge for data analysis. We review the application of multivariate data analysis and pattern recognition techniques to flow cytometry. METHODS: Approaches were divided into two broad types depending on whether the aim was identification or clustering. Multivariate statistical approaches, supervised artificial neural networks (ANNs), problems of overlapping character distributions, unbounded data sets, missing parameters, scaling up, and estimating proportions of different types of cells comprised the first category. Classic clustering methods, fuzzy clustering, and unsupervised ANNs comprised the second category. We demonstrate the state of the art by using AFC data on marine phytoplankton populations. RESULTS AND CONCLUSIONS: Information held within the large quantities of data generated by AFC was tractable using ANNs, but for field studies the problem of obtaining suitable training data needs to be resolved, and coping with an almost infinite number of cell categories needs further research. PMID- 11429771 TI - Comparison of five clustering algorithms to classify phytoplankton from flow cytometry data. AB - BACKGROUND: Artificial neural networks (ANNs) have been shown to be valuable in the analysis of analytical flow cytometric (AFC) data in aquatic ecology. Automated extraction of clusters is an important first stage in deriving ANN training data from field samples, but AFC data pose a number of challenges for many types of clustering algorithm. The fuzzy k-means algorithm recently has been extended to address nonspherical clusters with the use of scatter matrices. Four variants were proposed, each optimizing a different measure of clustering "goodness." METHODS: With AFC data obtained from marine phytoplankton species in culture, the four fuzzy k-means algorithm variants were compared with each other and with another multivariate clustering algorithm based on critical distances currently used in flow cytometry. RESULTS: One of the algorithm variants (adaptive distances, also known as the Gustafson--Kessel algorithm) was found to be robust and reliable, whereas the others showed various problems. CONCLUSIONS: The adaptive distances algorithm was superior in use to the clustering algorithms against which it was tested, but the problem of automatic determination of the number of clusters remains to be addressed. PMID- 11429772 TI - Detection of subgroups from flow cytometry measurements of heterotrophic bacterioplankton by image analysis. AB - BACKGROUND: Flow cytometry is an invaluable tool for the analysis of large series of samples in aquatic microbial ecology. However, analysis of the resulting data is often inefficient or does not reflect the complexity of natural communities. Because bacterioplankton assemblages frequently fall into several clusters with respect to their cellular properties, these subgroups seem to be a promising level of abstraction. Image analysis was used to detect clusters from flow cytometry data. The method was tested on a bacterial community under heavy protozoan grazing pressure. METHODS: A bivariate histogram of flow cytometry data was transformed into a gray-scale image for image analysis. After low-pass filtration, regional maxima were delimited by a watershed algorithm. The resulting areas were then used as gates on the original measurements. RESULTS: Three clusters could be detected from the bacterial assemblage. Protozoan grazing had a strong impact on the bacterial community, which could be analyzed in detail at the level of individual subgroups. CONCLUSIONS: Investigation at the level of bacterial subgroups allowed a more detailed analysis than whole-community statistics and delivered essential and ecologically meaningful information. Image analysis proved to be an adequate tool to detect the subgroups without a priori knowledge. PMID- 11429773 TI - Flow cytometric analysis of chronic and acute toxicity of copper(II) on the marine dinoflagellate Amphidinium carterae. AB - BACKGROUND: Copper(II) is a heavy metal whose levels have increased in some marine ecosystems to polluting levels. Dinoflagellates, an important phytoplankton group, are at the base of aquatic food chains and bioaccumulation of copper by these microorganisms can result in complex ecosystem alterations, so we investigated how copper disturbs those cells. METHODS: Cytotoxic effects of sublethal and lethal copper concentrations ranging from 4.2 nM (control condition) to 3.13 microM estimated labile copper were studied in batch cultures of Amphidinium carterae. Cell morphology, motility, autofluorescence, and fluorescein diacetate (FDA)-dependent fluorescence generation were evaluated by flow cytometry (FCM) and microscopy. RESULTS: Exposure of A. carterae to toxic levels of copper impaired cell mobility, delayed cell proliferation, led to increased green autofluorescence, and at 3.13 microM labile copper also induced encystment and death. Chlorophyll fluorescence, however, was not affected. Kinetic FCM assay of FDA-dependent fluorescence generation showed a dose dependent enhancement of fluorescein fluorescence immediately after copper addition and in cultures with sustained exposure to this toxicant. CONCLUSIONS: Our data suggest that copper toxicity occurs quickly at the membrane level in relation to oxidative stress generation. Based on fluorescence kinetic studies, the Na(+)/H(+) antiporter seemed to be affected by copper, thereby affecting intracellular pH. PMID- 11429774 TI - Monitoring phytoplankton, bacterioplankton, and virioplankton in a coastal inlet (Bedford Basin) by flow cytometry. AB - BACKGROUND: To establish the prevailing state of the ecosystem for the assessment of long-term change, the abundance of microbial plankton in Bedford Basin (Nova Scotia, Canada) is monitored weekly by flow cytometry. METHODS: Phytoplankton are detected by their chlorophyll autofluorescence. Those that contain phycoerythrin are designated as Synechococcus cyanobacteria or cryptophyte algae according to the intensity of light scatter. Bacteria and viruses are stained with DNA-binding fluorochromes and detected by green fluorescence. Distinction is made between bacterial and viral subpopulations exhibiting high and low fluorescence. RESULTS: Time series data are presented for weekly observations from 1991 to 2000. Weekly averages are computed for the complete annual cycle of temperature, salinity, river discharge, nitrate, phosphate, silicate, chlorophyll, total phytoplankton including Synechococcus and cryptophytes, total bacteria including high and low fluorescence subpopulations, and total viruses including high and low fluorescence subpopulations. CONCLUSIONS: The microbial biomass in the surface water of Bedford Basin is dominated by phytoplankton. The spring bloom of phytoplankton represents a maximum in algal biovolume, but not in cell number. Phytoplankton, bacteria, and viruses all attain their annual numerical maxima between the summer solstice and the autumn equinox. A vigorous microbial loop and viral shunt is envisioned to occur in the summer. PMID- 11429775 TI - Phytoplankton group dynamics in the Bay of Marseilles during a 2-year survey based on analytical flow cytometry. AB - BACKGROUND: The Bay of Marseilles is under the influence of a large urban concentration and its maritime activities. All of them discharge compounds (hydrocarbons, excess nutrients, heavy metals, chemicals, etc.) that can alter the marine ecosystem. To investigate whether ultraphytoplankton (<10 microm) could be used as biosensors for their own ecosystem, a 2-year survey was conducted in the Bay of Marseilles. METHODS: Seven stations monitored water mass and potential anthropic effects in the bay. Seawater samples were collected monthly or bimonthly at three depths, prefiltered, fixed, and kept in liquid nitrogen until flow cytometric analysis. RESULTS: Five categories were created: Prochlorococcus, picoeukaryotes (<2 microm), nanoeukaryotes I (2--6 microm), nanoeukaryotes II (6--10 microm), and Synechococcus (<1.5 microm). Artificial neural network analysis (Kohonen self-organizing maps) produced the same number of clusters as cluster analysis with Winlist software (Verity Software House). CONCLUSIONS: In addition to the wide variabilities in abundance and biomass, there were a strong seasonal signal and sporadic events. Lessons are derived from this study for future monitoring of marine microorganisms. PMID- 11429776 TI - Growth kinetics of algal populations exsymbiotic from Paramecium bursaria by flow cytometry measurements. AB - BACKGROUND: The ciliate Paramecium bursaria normally exists as a green paramecium system because each animal cell carries several hundred, unicellular, green, algal cells in its cytoplasm. One of the remarkable and poorly understood pecularities of this system is the steady state in the number of algae per protozoan cell. A major point in the study of mechanisms governing the persistence of symbiont numbers is adequate understanding of the algal life cycle. METHODS: Asynchronously growing cell populations of several algal strains (SA-1, SA-3, and SA-9) exsymbiotic from P. bursaria were characterized by flow cytometry. Algal endogenous chlorophyll and DNA contents were monitored to analyze cell growth kinetics at logarithmic and stationary culture phases. Cell sorting visualized the morphology of algae corresponding to the hyperhaploid (2C and 4C) DNA peaks. RESULTS: Cell-division cycle-dependent changes in chlorophyll and DNA content distributions were most dramatic in logarithmically growing algal populations (an increase in the number of S-phase cells and cells with more chlorophyll), which are thought to be associated with accelerated DNA and chlorophyll metabolism in log-phase algal cultures. Upon reaching the stationary phase of growth, algal populations distinctly showed, in addition to one haploid (1C) DNA peak, two hyperhaploid peaks (2C and 4C) corresponding mainly to cells with two and four nuclei, respectively. CONCLUSIONS: Growth characteristics of algae exsymbiotic from P. bursaria monitored by flow cytometry provide valuable information for the analysis of the algal life cycle, which is important for understanding the regulation mechanisms of symbiont numbers. PMID- 11429777 TI - Flow cytometry of cell proliferation through the incorporation of bromodeoxyuridine as an index of growth rate in the water flea, Daphnia magna (Crustacea, Cladocera). AB - BACKGROUND: In this paper, we used a small crustacean as a model to develop a method for quantifying growth rates through the measurement of a cell proliferation marker. This was done in order to study the feasibility of this assay for estimating zooplankton production in the ocean. Flow cytometry immunodetection of bromodeoxyuridine (BrdU) was performed to detect and quantify the cycling nuclei of Daphnia magna. METHODS: A combination of mechanical dissociation and cell enucleation procedures proved to be the most convenient method for preparing nuclear suspensions from whole organisms. Up to three populations of nuclei with different ploidy were observed. The relative abundance of these nuclear populations changed with the size of the flea. RESULTS: The staining technique has been optimized. The time and concentration for the maximum detection of BrdU-labeled nuclei were 3 h at 300 microM BrdU. Whole organisms can be frozen (-20 degrees C) after incubation with no changes in the final results. The method was used in different physiological conditions under controlled food and temperature in order to test the inverse relationship between physiological rates and size of organisms at several developmental stages. The quantification of BrdU-labeled nuclei in 1-6 day-old larvae showed the highest labeling index, with a mean of 95 +/- 1% (n = 22). In contrast, young animals (0.8-1.2 mm) had 25 +/- 4% (n =16, P < 0.001) and adults (>1.4mm) had 14 +/- 3% (n = 4, P < 0.001). The results obtained show an expected tendency, suggesting that a direct relationship exists between the labeling index and the instantaneous growth rate. CONCLUSIONS: Certain features of our method, such as the short times required for labeling and the possibility of preserving the samples during field experiments and under different conditions (including natural concentrations and types of food), are advantageous to the study of processes governing energy fluxes in pelagic ecosystems. PMID- 11429778 TI - Non-invasive treatments of luteinizing hormone-releasing hormone for inducing spermiation in American (Bufo americanus) and Gulf Coast (Bufo valliceps) toads. AB - As many as 20% of all assessed amphibian species are threatened with extinction, and captive breeding programs are becoming important components of conservation strategies for this taxon. For some species, exogenous hormone administration has been integrated into breeding protocols to improve propagation. However, most treatments are administered by an intraperitoneal injection that can be associated with some risks. The general goal of this study was to identify a non invasive method of applying luteinizing hormone-releasing hormone (LHRH), which reliably induces sperm release in toads. Specific objectives were to 1) test the spermiation response after topical application of different LHRH doses to the abdominal seat region, 2) evaluate the effects of adding the absorption enhancers dimethyl sulfoxide (DMSO), acetone, and glyceryl monocaprylate (GMC) to the LHRH, 3) assess the spermiation response after oral delivery of LHRH in a mealworm vehicle, and 4) compare sperm characteristics and spermiation responses to treatments in two different toad species. Male American (n = 9) and Gulf Coast (n = 7) toads were rotated systematically through a series of treatments. Urine was collected and evaluated for the presence of sperm at 0, 3, 7, 12, and 24 hours post-treatment. There were no statistical differences in spermiation induction or sperm characteristics between American and Gulf Coast toads after the treatments. Oral administration of 100 &mgr;g LHRH was occasionally successful in inducing spermiation, but results appeared largely unreliable. Ventral dermal application of 100 or 10 &mgr;g LHRH in 40% DMSO were more effective (P < 0.05) at inducing spermiation compared with the other treatments tested, eliciting sperm release in more than 70% of toads tested. In breeding programs for rare and/or fragile anurans, these non-invasive methods of exogenous hormone administration might be preferred over intraperitoneal injections. Zoo Biol 20:63-74, 2001. Copyright 2001 Wiley-Liss, Inc. PMID- 11429779 TI - Lipid composition, fatty acid profiles, and lipid-soluble antioxidants of eggs of the Hermann's tortoise (Testudo hermanni boettgeri). AB - The major lipid classes, their fatty acid profiles, and the amounts of the lipid soluble components, vitamin E, vitamin A, and carotenoids, were determined for egg yolks of the Hermann's tortoise (Testudo hermanni boettgeri) with the aim of identifying any features that may potentially impair the adaptation of this endangered species to deteriorations in habitat. Total lipid formed 16% (wt/wt) of the fresh yolk and consisted of (wt/wt) 74.4% triacylglycerol, 18.1% phospholipid, 3.0% cholesteryl ester, and 3.4% free cholesterol. Despite a diet based on green plants, contributing alpha-linolenic acid as the main polyunsaturate, this fatty acid formed only 3.8% of the total mass of fatty acid of the total lipid. The main acyl component of the yolk lipids was the monounsaturated fatty acid, oleic acid, which formed 45.6% of the total. The most striking feature of the yolk composition was the almost complete lack of two nutrients, docosahexaenoic acid and vitamin A, which are essential for the developing embryo. Although it is feasible that the embryo synthesizes docosahexaenoic acid from yolk-derived alpha-linolenic acid and also converts yolk-derived beta-carotene to vitamin A, the yolk is poorly endowed with both these precursors. The stringencies displayed by the yolk composition in this species may limit the flexibility to adapt to changes in the availability of food items when the habitat is threatened. Zoo Biol 20:75-87, 2001. Copyright 2001 Wiley-Liss, Inc. PMID- 11429780 TI - Disease risk and inter-institutional transfer of specimens in cooperative breeding programs: Herpes and the elephant species survival plans. AB - Managers of cooperative breeding programs and re-introduction projects are increasingly concerned with the risk of disease transmission when specimens are transferred among facilities or between facilities and the natural environment. We used data maintained in North American studbooks to estimate the potential risks of disease transmission by direct and indirect contact of specimens in the American Zoo and Aquarium Association's Elephant Species Survival Plan. Histological evidence for a novel herpesvirus disease transmitted between and within elephant species housed in North American facilities prompted an examination of the scope of possible transmission routes within the captive population. We found that, compared with other species managed through Species Survival Plans, elephants experience relatively few transfers between zoos. Nevertheless, the number of direct contacts with other elephants born during the study period of 1983-1996 (excluding stillbirths) was much higher than we had anticipated (&mgr; = 25 +/- 27; N = 59) and the number of potential indirect contacts was surprisingly large (&mgr; = 143 +/- 92; N = 59). Although these high rates of potential contacts complicate exact identification of infection pathways for herpesvirus, we were able to propose potential routes of transmission for the histologically identified cases. Furthermore, the extraction of data from studbooks allowed us to readily identify other specimens that did not succumb to the disease despite similar exposure. Moreover, we were able to identify other possible cases to recommend for histological examination. Herein we reveal the possibilities of multiple disease transmission pathways and demonstrate how complex the patterns of transmission can be, confounded by the unknown latency of this novel herpesvirus. This emphasizes the need for zoo veterinarians and cooperative breeding programs to consider the full potential for disease transmission associated with each and every inter-zoo transfer of specimens. Zoo Biol 20:89-101, 2001. Copyright 2001 Wiley-Liss, Inc. PMID- 11429781 TI - Reproductive steroid hormones and ovarian activity in felids of the Leopardus genus. AB - Reproductive endocrine patterns were characterized in female ocelots (Leopardus pardalis; n = 3), tigrinas (Leopardus tigrinus; n = 2), and margays (Leopardus wiedii; n = 2) housed in captivity in southern Brazil. Females were maintained as singletons and exposed to natural fluctuations in photoperiod. Cyclic changes in ovarian steroids were monitored by analyzing estrogen and progestogen metabolites in fecal samples collected five times weekly for 14 to 18 months. Based on intervals between fecal estrogen peaks, mean (+/- SEM) duration of the estrous cycle was 18.4 +/- 1.6 days for the ocelots (range, 7-31 days; n = 75 cycles), 16.7 +/- 1.3 days for the tigrinas (range, 11-27 days; n = 23 cycles), and 17.6 +/- 1.5 days for the margays (range, 11-25 days; n = 32 cycles). Fecal progestogen analyses combined with two laparoscopic observations of the ovaries confirmed that ocelots and tigrinas did not ovulate spontaneously. In contrast, non-mating-induced luteal phases of 40.1 +/- 6.3 days in duration (range, 30-60 days) were observed frequently in both margays. There was no evidence of gonadal seasonality in margays in either follicular or luteal activity. In ocelots, cyclic changes in estrogen excretion were observed during each month of the year; however, only one female cycled continuously. In the other two ocelots, periods of acyclicity of several months' duration were observed. It was not possible to conclude whether tigrinas were aseasonal because estrous cyclicity was observed in only one of two individuals. In the female that cycled, a 3-month period of acyclicity was observed in the late fall/early winter. These data demonstrate similarities among three felid species of the genus Leopardus, including evidence they are polyestrous but experience unexplained periods of ovarian inactivity. Only the margays differed by exhibiting occasional spontaneous, non-mating induced ovulations. Historically, these species have not bred well in captivity. However, it is hoped that understanding the biological similarities and differences among them could lead to improved management strategies that may one day result in increased reproductive success. Zoo Biol 20:103-116, 2001. Copyright 2001 Wiley-Liss, Inc. PMID- 11429782 TI - Induction of the SAPK activator MIG-6 by the alkylating agent methyl methanesulfonate. AB - The alkylating agent methylmethanesulfonate (MMS) activates the c-jun N-terminal kinase (JNK)/stress-activated protein kinase (SAPK) and the p38 mitogen-activated protein kinase (p38MAPK) pathways via different mechanisms of action. Activation of p38MAPK by MMS involves the pp125 focal adhesion kinase-related tyrosine kinase RAFTK and the MAPK kinase 3. The way in which MMS can activate JNK/SAPK has not been elucidated. Here we describe the identification by differential display of human mitogen-activated gene-6 (MIG-6) as a novel MMS-inducible gene. Induction of MIG-6 by MMS was found in human diploid skin fibroblasts and in simian virus 40-transformed skin fibroblasts, indicating that the enhanced expression of MIG-6 after MMS-treatment did not require p53. The signal leading to activation of MIG-6 appeared to be independent of DNA damage. High MIG-6 expression was found in the liver, lung, and placenta. MIG-6 is an adapter protein that binds to the activated form of cdc42Hs and to 14-3-3 proteins, thereby activating JNK/SAPKs. Our results suggest that activation of JNK/SAPKs by MMS may involve the induction of MIG-6. PMID- 11429783 TI - CTNNB1 mutations and beta-catenin protein accumulation in human hepatocellular carcinomas associated with high exposure to aflatoxin B1. AB - beta-Catenin plays a key role in the Wnt signaling pathway, and mutations of CTNNB1, the gene that encodes beta-catenin, have been identified in about one fourth of human hepatocellular carcinomas from regions of low aflatoxin B1 exposure. In this study 62 hepatocellular carcinomas (HCCs) from people highly exposed to aflatoxin B1 in Guangxi, People's Republic of China, were laser capture microdissected and examined for CTNNB1 mutations. In addition, 41 of the HCCs were evaluated for the presence of the beta-catenin protein by immunohistochemical methods. Twenty of the HCCs showed positive results for beta catenin, with strong membrane staining, while adjacent non-neoplastic liver tissue lacked or showed only weak membrane staining. One HCC, in which a CTNNB1 mutation was not detected, showed nuclear staining for the beta-catenin protein. Mutations of CTNNB1 were identified in five HCCs. These consisted of four point mutations in the glycogen serine kinase-3beta phosphorylation region of codons 32 45 and one deletion of codons 32-38. These mutations were similar to those previously reported for human HCC, although at a lower frequency. A signature mutation profile associated with aflatoxin B1 exposure could not be identified. The immunohistochemical findings indicate a role for accumulation of beta-catenin and possibly increased Wnt signaling in aflatoxin B1-associated HCC. The low frequency of CTNNB1 mutations, however, suggests that mutation of another Wnt signaling component, such as the Wnt scaffolding protein axin or the adenomatous polyposis coli protein, both of which modulate beta-catenin stability, also may be involved in aflatoxin-associated HCC. Published 2001 Wiley-Liss, Inc. PMID- 11429784 TI - Role of matrix metalloproteinase-9 in progression of mouse skin carcinogenesis. AB - Invasion of malignant tumor cells is required for the formation of metastatic colonies. Uncontrolled expression of matrix metalloproteinase (MMP)-2 and MMP-9 is a critical part of the invasive potential of tumor cells and is affected by the balance between the enzymes and the inhibitors secreted by the cell. Here we analyzed the expression and activity of the two gelatinases (MMP-2 and MMP-9) as well as the expression levels of the tissue inhibitor of metalloproteinase (TIMP2)-, in different stages of carcinogenesis using mouse skin cell lines derived from tumors induced by chemical mutagens. Our results suggested that the expression of MMP-9 was implicated in the progression to spindle cell carcinomas in mouse keratinocytes. MMP-2 levels remained steady in all cell lines, whereas levels of TIMP-2 were increased in normal and spindle cells. The AP-1 DNA binding and transcriptional activity on the MMP-9 promoter were increased in the malignant cell lines, indicating the requirement of this binding site for its activation. The results of this study clearly suggested the important role of MMP 9, but not of MMP-2, in the metastatic properties of mouse keratinocytes. PMID- 11429785 TI - Suppression of cell transformation and induction of apoptosis by caffeic acid phenethyl ester. AB - Caffeic acid phenethyl ester (CAPE), which is derived from the propolis of honeybee hives, has been shown to block tumor promotion and to have toxic effects on several cancer cells. The mechanism of the anti-tumor promotion activity of CAPE is unclear, however. In this study, we found that CAPE suppressed 12-O tetradecanoylphorbol-13-acetate-induced cell transformation and induced apoptosis in mouse epidermal JB6 Cl 41 cells. No difference in induction of apoptosis was observed between normal lymphoblasts and sphingomyelinase-deficient cell lines. Although CAPE treatment of two p53 mutant tumor cell lines, NCI-H358 and SK-OV-3, and p53-deficient (p53(-/-)) cells caused the cleavage of caspase-3 as well as DNA fragmentation, caspase-3 cleavage was seen early (at 6 h) only in cells expressing wild-type p53 (p53(+/+)) and Cl 41 cells. These results suggested that p53 may be involved in the early stage of CAPE-induced apoptosis. The p53 dependent transcription activation occurred 2 h after treatment with CAPE and reached a maximum at 6 h in Cl 41 p53 DNA-binding sequence stable transfectant cells. In addition, phosphorylation of p53 at serine 15 and serine 392 was induced in Cl 41 cells within 6 h after treatment with CAPE. Therefore, CAPE may induce apoptosis through p53-dependent and p53-independent pathways and its anti tumor promotion activity may have occurred through the induction of apoptosis. PMID- 11429786 TI - Deregulated expression of DP1 induces epidermal proliferation and enhances skin carcinogenesis. AB - E2F transcription factors have been implicated in several cellular processes, including proliferation, apoptosis, and oncogenic transformation. A functional E2F factor consists of a heterodimer containing an E2F polypeptide (E2F1-E2F6) and a DRTF1-polypeptide (DRTF1-polypeptide-1 (DP1) or DRTF1-polypeptide-2). It is the E2F subunit that supplies the transcriptional activation domain and the motif involved in binding to members of the retinoblastoma tumor suppressor family. The role of the DP subunit in regulating E2F-dependent activities is not completely understood. To examine the properties of DP1 in vivo, we generated transgenic mouse lines expressing DP1 under the control of a keratin 5 (K5) promoter. Overexpression of DP1 in basal layer keratinocytes caused mild hyperplasia and hyperproliferation of the epidermis but did not result in increased apoptosis or spontaneous tumor development. Coexpression of DP1 with E2F1 or E2F4 in the epidermis of bigenic mice modestly enhanced proliferation and apoptosis over the levels induced by E2F1 or E2F4 expression alone. In a two-stage chemical carcinogenesis assay, more and larger skin tumors developed in K5 DP1 transgenic mice than in nontransgenic mice. These findings show that in this in vivo model, deregulated expression of DP1 on its own induced proliferation and enhanced carcinogenesis. PMID- 11429787 TI - Regulation of cellular invasion and matrix metalloproteinase activity in HepG2 cell by connexin 26 transfection. AB - We previously reported that connexin (Cx) 26 expression is involved in negative growth control of HepG2 cells established from a human hepatoma. We also found that induction of E-cadherin and subsequent formation of a cell adhesion complex were induced in HepG2 cells by Cx 26 expression. To examine the exact role of Cx 26-induced E-cadherin junctions in regulating appearance of malignant phenotypes of HepG2 cells, we expressed a Cx 26 antisense oligodeoxynucleotide (AS-ODN) in an established HepG2 cell clone that has stable expression of Cx 26 genes. We investigated changes in the expression of E-cadherin, the localization of beta catenin, and some malignant phenotypes of HepG2 clone after the suppression of Cx 26 expression by AS-ODN treatment. The AS-ODN treatment prevented the expression of Cx 26 and E-cadherin, and the localization of beta-catenin was changed from cytoplasmic membrane to the cytoplasm. In parallel, a morphological change from a monolayer of polygonal cells to multilayered colonies was induced by the treatment, indicating a change of a malignant phenotype of HepG2 cells. The activity of matrix metalloproteinase 9 (MMP-9) was elevated by the AS-ODN treatment. A concomitant increase in invasiveness of the Cx 26-expressing cells by the treatment was also observed in an in vitro assay with Matrigel matrix. These results suggest that the induction of E-cadherin and formation of the cell adhesion complex by Cx 26 expression contribute to the reversal of some malignant phenotypes of HepG2 cells. Furthermore, the Cx 26-dependent expression of E cadherin leads to reduction of the invasiveness of the cells through suppression of MMP-9 activity. PMID- 11429801 TI - Synthesis of 2'-modified oligodeoxynucleotides via on-column conjugation. AB - Oligodeoxynucleotides modified at the 2'-position of 2'-amino-2'-deoxyuridine or uridine were prepared in high yield and purity using phosphoramidites 2 and 3, respectively. Oligodeoxynucleotide conjugates were prepared on the solid-phase synthesis support following selective unmasking of the nucleophile incorporated in these phosphoramidites. Synthesis of oligodeoxynucleotides modified at the 2' position of an internal nucleotide provides molecules that are complementary to those previously prepared via a similar approach using C5-substituted pyrimidines. The efficiency of functionalization of the 2'-O-alkylamino-uridine derived from 3 in a protected oligodeoxynucleotide was less susceptible to steric hindrance than the 2'-amino-2'-deoxyuridine in the same polymeric substrate. However, the greater reactivity of the 2'-O-alkylamine containing nucleotide gave rise to undesired acetamide formation resulting from nucleophilic attack on the 5'-terminal acetate in capped failure sequences. This problem was overcome by using 2,2,2-trimethylacetyl anhydride as a capping agent during the automated synthesis cycles. Finally, the efficiency of the photochemical unmasking of the support bound alkylamine on a 1 mumole scale was improved by using two 20 min photolysis cycles, coupled with removing reaction byproducts between cycles. PMID- 11429802 TI - An old but simple and efficient method to elucidate the oxidation mechanism of NAD(P)H model 1-Aryl-1,4-dihydronicotinamides by cations 2-methyl-5 nitroisoquinolium, tropylium, and xanthylium in aqueous solution. AB - Cations 2-methyl-5-nitroisoquinolinium (IQ+), tropylium (T+), and xanthylium (Xn+) were treated by an NAD(P)H model 1-(p-substituted phenyl)-1.4 dihydronicotinamide series (1) in buffered aqueous solution to give the corresponding reduced products by accepting hydride. Effects of the 4 substituents of 1 on the reaction rates were investigated. Hammett's linear free energy relationship analysis on the three reactions of 1 provides the reaction constants of -0.48, -2.2, and -1.4 with IQ+, T+, and Xn+ as the hydride acceptors, respectively. Comparison of the present reactions with the reaction examples whose mechanisms are well-known, such as the reaction of 1 with a one electron oxidant Fe(CN)6(-3), shows that the active site of 1 in the oxidation with IQ+ is at the 4-position on the dihydropyridine ring but that the active site of 1 in the oxidations with T+ and Xn+ is at the 1-position, which is in agreement with the results from the Bronsted-type linear analysis and the relation studies of the logarithm of the second-order rate constants with the oxidation potentials of the hydride donors. According to the dependence of the reaction mechanism on the active site of 1, a conclusion can be made that the reaction of 1 with IQ+ proceeds by direct one-step hydride transfer mechanism, but the reactions of 1 with T+ and Xn+ would take place via multistep hydride transfer mechanism initiated by one-electron transfer. PMID- 11429803 TI - Nature of the 2-bicyclo[3.2.1]octanyl and 2-bicyclo[3.2.2]nonanyl cations. AB - Density functional and ab initio calculations have been employed to explore the nature of the cations formed in the solvolysis of 2-bicyclo[3.2.1]octanyl tosylate the 2-bicyclo[3.2.2]nonanyl tosylate. In contrast to recent conclusions in the literature, the various proposed classical carbocations postulated to explain the products of the 2-bicyclo[3.2.2]nonanyl tosylate solvolysis were found to have nonclassical structures. PMID- 11429804 TI - Nitroso group transfer from substituted N-methyl-N-nitrosobenzenesulfonamides to amines. Intrinsic and apparent reactivity. AB - We have studied the nitroso group transfer from substituted N-methyl-N nitrosobenzenesulfonamides to primary and secondary amines, observing that the rate of the reaction increases as a consequence of the presence of electron withdrawing groups on the aromatic ring of the nitrosating agents. The rate constants determined for the nitroso group transfer, ktr, give good Bronsted-type relationships between log ktr (rate constant for nitroso group transfer) and pKaR2NH2+ and pKaleaving group. The study of the nitrosation processes of secondary amines catalyzed by ONSCN and denitrosation catalyzed by SCN-, in combination with the formation equilibrium of ONSCN, has enabled us to calculate the value of the equilibrium constant for the loss of the NO+ group from a protonated N-nitrosamine (pKNOR2N+HNO), which can be defined by analogy with pKaR2NH2+. The value of pKNOX-NO for the loss of the NO+ group from an N-methyl-N nitrosobenzenesulfonamide was obtained in a similar way. By using values of delta pKNO = pKNOR2N+HNO - pKNOX-NO, we were able to calculate the equilibrium constant for the nitroso group transfer and characterize the transition state. On the basis of Bronsted-type correlations, we have obtained values of beta nuclnorm and alpha lgnorm approximately equal to 0.55, showing a perfectly balanced transition state. In terms of the Marcus theory, the calculation of the intrinsic barriers for the nitroso group transfer reaction shows that the presence of electron withdrawing groups on the aromatic ring of the N-methyl-N nitrosobenzenesulfonamides does not cause these barriers to vary. PMID- 11429805 TI - Lamellar organic thin films through self-assembly and molecular recognition. AB - Molecular clips possessing U-shaped cavities have been functionalized on their convex side with long aliphatic tails. These molecules form dimers which self assemble into malleable lamellar thin films. Upon addition of a guest (methyl 3,5 dihydroxybenzoate), a 1:1 host-guest complex is formed, which prohibits clip dimerization. As a result, the lamellar structure of the material is lost. Complexation of 3,5-dihydroxybenzoic acid in the clip results in host-guest complexes which dimerize by hydrogen bonding interactions between the carboxylic acid functions of the bound guests. This dimerization restores the lamellar type architecture of the material. PMID- 11429806 TI - Friedlander synthesis of chiral alkyl-substituted 1,10-phenanthrolines. AB - The synthetic scope of the Friedlander condensation in the preparation of chiral alkyl-substituted 1,10-phenanthrolines has been investigated. A range of chiral [x,y-b]-cycloalkeno-condensed phenanthrolines has been prepared in one step from steroidal or other cyclic ketones from the chiral pool and 8-amino-7 quinolinecarbaldehyde (1) via base-catalyzed condensation. Phenanthroline derivatives are formed in good yields with unhindered ketones, but the reaction proceeds even with sterically congested substrates such as camphor, albeit in low yield. The utility of the Friedlander condensation has been extended to the synthesis of chiral 3-alkyl-substituted phenanthrolines from monoalkyl substituted acetaldehydes. PMID- 11429807 TI - Titanium(IV) chloride and quaternary ammonium salt promoted Baylis-Hillman reaction. AB - The Baylis-Hillman reaction of aldehydes with alpha,beta-unsaturated ketones can be drastically affected by the reaction temperature and Lewis bases. When the reaction was carried out at -78 degrees C using catalytic amounts of quaternary ammonium salts (R4N+X-, X = Cl, Br, I) as Lewis bases, in the presence of titanium(IV) chloride, the chlorinated aldol adduct 1 was obtained as the major product. Quaternary ammonium bromides and iodides (R4N+X-, X = Br, I) have higher catalytic activity than corresponding chlorides (R4N+Cl-). Quaternary ammonium fluorides (R4N+F-) do not have activity at all. The amounts of Lewis acid and quaternary ammonium salts used affect the reaction rate and product. A plausible reaction mechanism is proposed. If the reaction was carried out at room temperature (about 20 degrees C) in the presence of titanium(IV) chloride and quaternary ammonium salts (R4N+X-, X = Cl, Br, I), the elimination product 3, derived from 1, was formed as the major product. PMID- 11429808 TI - Synthesis of isoindolo[2,1-alpha]indoles by the palladium-catalyzed annulation of internal acetylenes. AB - A wide variety of substituted isoindolo[2,1-alpha]indoles have been prepared via annulation of internal alkynes by imines derived from o-iodoanilines in the presence of a palladium catalyst. This methodology provides an extremely efficient route for the synthesis of these tetracyclic heterocycles from readily available starting materials. The mechanism of this interesting annulation process appears to involve (1) oxidative addition of the aryl iodide to Pd(0), (2) alkyne insertion, (3) addition of the resulting vinylic palladium intermediate to the C-N double bond of the imine, (4) either electrophilic palladation of the resulting sigma-palladium intermediate onto the adjacent aromatic ring derived from the internal alkyne or oxidative addition of the neighboring aryl carbon-hydrogen bond, and (5) reduction of the tetracyclic product and Pd(0). A variety of internal acetylenes have been employed in this annulation process in which the aromatic ring of the alkyne contains either a phenyl or a heterocyclic ring. PMID- 11429809 TI - Acylation and related reactions under microwaves. 4. Sulfonylation reactions of aromatics. AB - Solvent-free sulfonylation of benzene and its activated or deactivated derivatives were carried out under microwave (MW) irradiation and a catalytic amount of iron(III) chloride, which, under these conditions, is more active than other metallic salts. With more reactive and/or nonvolatile reagents (anisole, xylenes, mesitylene) expeditious conditions (short reaction time at constant MW power without control of the temperature) were used. With less reactive and/or low-boiling reagents (benzene, toluene, halobenzenes), the rise in temperature and the increase of reaction time were controlled either by sequential MW irradiation or by a temperature order. It was shown that MWs cause preferential interactions with polar species present in the reaction, especially the aryl sulfone and its FeCl3-complexed form. A MW nonthermal effect was not observed when identical temperature gradients were produced by classical heating and MW irradiation, and if reaction temperature was strictly controlled. PMID- 11429810 TI - Synthesis and autoxidation of new tetracyclic 9H,10H-indolizino[1,2-b]indole-1 ones. AB - The new tetracyclic 9H,10H-indolizino[1,2-b]indole-1-one derivatives (7a-d, 7ea, 7eb) have been synthesized by modified Fischer indole synthesis from the enol ether of 2,5-dihydroxy-7-methyl-6-cyano-indolizine (3) and arylhydrazines (4a-g). Attempted N-methylation of 7a-d produced a series of autoxidized products including 10-hydroperoxy-1-methoxyindolizino[1,2-b]indole (9a-d) as the major product accompanied with methylperoxides (10a-d and 11a-d) and 2-formyl-3 (pyridine-2-yl)indole (12a, 12c) derivatives as the minor products. A plausible mechanism of the autoxidation is postulated based on the isolation of some intermediates. The reaction is thought to proceed through azaenolate/enamine intermediates following a novel type of autoxidation. PMID- 11429811 TI - Mechanism of the addition reaction of alkyl azides to [60]fullerene and the subsequent N2 extrusion to form monoimino-[60]fullerenes. AB - The 1,3-dipolar cycloaddition of methyl azide to C60 and the subsequent nitrogen elimination from the formed triazoline intermediate to yield the aziridine adduct have been studied using semiempirical and density functional methods. The results obtained show that the addition of methyl azide to C60 takes place in the ring junction between two six-membered rings leading to a closed [6,6]-trizoline intermediate with an energy barrier of about 20 kcal mol-1 and an exothermicity of ca. 2 kcal mol-1 at the B3LYP/6-31G**//AM1 level of theory. The subsequent thermal loss of N2 takes place through a stepwise mechanism in which the cleavage of the N-N single bond precedes the breaking of the N-C bond, with a total activation energy of approximately 45 kcal mol-1. The N2 loss occurs simultaneously with the formation of the new N-C bond. During the process, the steric effects of the leaving N2 molecule prevent the addition of the nitrene substituent to the [6,6]-ring junction attacked initially and force the addition to an adjacent [5,6]-ring junction. PMID- 11429812 TI - Chemoselective acylation of fully deprotected hydrazino acetyl peptides. Application to the synthesis of lipopetides. AB - Fully deprotected N-terminal alpha-hydrazino acetyl peptides were synthesized and chemoselectively acylated on the hydrazine moiety with various fatty acid succinimidyl esters or N-(cholesterylcarbonyloxy) succinimide to give lipopeptides of high purity. The buffer and pH were adjusted in order to minimize the oxidation of the hydrazine moiety and to achieve the best conversion and selectivity. The acylation was performed in a citrate-phosphate buffer/2 methylpropan-2-ol mixture of pH 5.1. The pKa of the alpha-hydrazino acetyl group on our model peptide was found to be 6.45, i.e., about 2 units lower than the pKa of a glycyl residue. The reaction was subsequently applied to the synthesis of a 38AA peptide derivatized by a palmitoyl group. PMID- 11429813 TI - Further studies of the Daphniphyllum alkaloid polycyclization cascade. AB - The scope of the 2-azadiene intramolecular Diels-Alder cyclization, previously employed for synthesis of the Daphniphyllum alkaloids, has been further investigated. Through a series of 1,5-diol cyclization precursors the substitution pattern of both the dienophile and the 2-azadiene were examined. From these studies it was shown that the cascade reaction is tolerant toward a variety of alkyl-substituted dienophiles. However, it was also demonstrated that this reaction is very sensitive to the substitution pattern of the 2-azadiene. Alterations made to the structure of the 2-azadiene cause either competing side reactions or complete failure of the reaction cascade. PMID- 11429814 TI - Poly-L-proline type II peptide mimics based on the 3-azabicyclo[3.1.0]hexane system. AB - This paper describes conformational studies of proline-templated amino acids (PTAAs) based on the 3-azabicyclo[3.1.0]hexane system as well as conformational studies on short peptides composed of these PTAAs. NOE data, coupling constants, and molecular modeling are consistent with a flattened boat conformation for monomeric and oligomeric residues based on this bicyclic system. NMR studies on dimeric and trimeric oligomers are consistent with a populated poly-L-proline type II conformation in CDCl3 and D2O. Solution studies and molecular modeling predicts phi approximately -70 degrees, psi approximately 131 degrees, chi 1 approximately -57 degrees, and chi 2 approximately -158 degrees for oligomeric residues. PMID- 11429815 TI - Synthesis of near-IR absorbing/emitting porphyrazine derivatives with tunable solubility. AB - We report the synthesis of porphyrazines (pzs), or tetraazaporphyrins, of the form H2[pz(An;B4-n)], where A is [S(CH2)3COOR]2 (R = n-Pr,H) and B is a fused beta,beta'-diisopropyloxybenzo group, including the compounds with n = 4 (6), n = 3 (7) and the trans compound with n = 2 (8) (Scheme 1). The synthesis employs Linstead crossover macrocyclization of dimethyl 6,7-dicyano-5,8-dithia-6(Z) dodecenedioate, MNT(C4O2Me)2 (2), with 1-imino-4,7-bis(1-methylethoxy)-1H isoindole-3-amine (4). These pigments were characterized by 1H NMR, 13C NMR, absorbance/fluorescence spectroscopy, mass spectrometry, and microanalysis. An X ray crystal structure of 8 is presented. Of particular note, 6-8 display intense near-IR absorbance and dual UV-visible/near-IR emission which are very important in potential biomedical applications, both for cancer therapy (photodynamic therapy, PDT) and cancer diagnosis (optical tumor imaging). For example, the trans-porphyrazine 8 has an intense long-wavelength absorption at ca. 800 nm (log epsilon = 4.18) and S1 fluorescence at approximately 820 nm, where mammalian tissue is effectively penetrated by light. Transformation of the ester group permits a wide range of functionality and solubility to be generated without change in optical properties. As an example, hydrolysis of these compounds by LiOH in THF/H2O gives the corresponding carboxylato-functionalized pigments 9-11, which are described. The last of these dissolves without aggregation in fetal calf serum. PMID- 11429816 TI - Computational study of radical cations of saturated compounds with sigma-type and pi-type N-N bonds. AB - Geometrical and electronic properties have been calculated and are compared with experimental data for three saturated diaza compounds and their radical cations and dications. The molecular geometries in the different oxidation states are consistently reproduced very well using the B3PW91 and B3LYP three-parameter density functional methods, with a modest 6-31G* basis set. The performance of the pure density functionals BLYP and BPW91 is less satisfactory. The Hartree Fock method yields excellent results in some cases but poor results in others. Ionization potentials and electron-nuclear hyperfine interactions are reproduced moderately well with B3LYP and B3PW91. Electronic excitation energies calculated with time-dependent density functional theory agree very well with experiment in most cases. For 2,7-diazatetracyclo[6.2.2.2(3,6).0(2,7)]tetradecane 2 and its radical cation and dication, the reorganization parameters for self-electron exchange were calculated and compared with experimental and earlier computed data. The calculations allow a good estimate of the different contributions to the energy barrier, i.e., the internal and solvent reorganization energies and the work term in the case of 2+/2++. PMID- 11429817 TI - Photophysics and photostereomutation of Aryl methyl sulfoxides. AB - The effect of a methanesulfinyl group on the photophysics of several aromatic chromophores is reported. The spectroscopic singlet and triplet energies are affected only modestly, compared to that parent arenes, but the fluorescence yields fall by at least 1 order of magnitude. Fluorescence lifetimes are short. Fluorescence enhancements are observed on cooling the sulfoxides from room temperature to 77 K. High quantum yields of stereomutation are reduced as the temperature drops. There is not a consistent effect on triplet or phosphorescence yields. It is proposed that these results are consistent with a nonradiative pathway for deactivation of the singlet that results in stereomutation. A modest activation energy of a few kcal/mol is estimated for the photochemical racemization of 1-methanesulfinylpyrene. PMID- 11429818 TI - Biaryl-based macrocyclic and polymeric chiral (salophen)Ni(II) complexes: synthesis and spectroscopic study. AB - Polymeric/oligomeric and macrocyclic (salophen)Ni(II) complexes have been synthesized starting from both an achiral biphenol dialdehyde and an optically active BINOL dialdehyde. It was found that these polysalophens contain nonplanar coordination of Ni(II) units that are paramagnetic. This is different from the previously reported (salophen)Ni(II) complexes which are square planar and diamagnetic. The nonplanar (salophen)Ni(II) units make the new polymeric Ni(II) complexes different from the helical structure proposed for chiral biaryl-based polymers containing square-planar (salophen)Ni(II) units. The copolymerization of the chiral binaphthyl monomer with the achiral biphenyl monomer demonstrates that the chirality of the binaphthyl unit is not propagated along the biphenyl polymer chain. PMID- 11429819 TI - Conformational studies by dynamic NMR. 78. Stereomutation of the helical enantiomers of trigonal carbon diaryl-substituted compounds: dimesitylketone, dimesitylthioketone, and dimesitylethylene. AB - The free energies of activation for the enantiomerization of the title compounds (Mes2C = X, Mes = 2,4,6-trimethylphenyl) were determined by dynamic NMR to be 4.6, 6.5, and 9.2 kcal mol-1 for X = O, S, and CH2, respectively. Single-crystal X-ray diffraction showed that the structure of dimesitylketone is that of a propeller (C2 symmetry) with the mesityl rings twisted by 50 degrees with respect to the plane of carbonyl. The same structure was predicted by molecular mechanics calculations, which also produced good agreement between computed and experimental barriers for a dynamic process where a disrotatory one-ring flip pathway reverses the helicity of the conformational enantiomers. Solid-state NMR spectra indicated that the enantiomerization barrier in the crystal must be much higher (at least 19 kcal mol-1) than that in solution. Contrary to the case of dimesitylketone, the calculated barrier of dimesitylethylene agrees better with the experimental value if the enantiomerization process is assumed to be a conrotatory two-ring flip pathway. PMID- 11429820 TI - BrF3, an underutilized reagent in organic chemistry: a novel C-C-N to C-N-C rearrangement. AB - Little is known about bromine trifluoride in organic chemistry. Under the right conditions, it can be a useful tool and generate a new and unprecedented chemistry. Thus, when reacted with oxime methyl ethers of alpha-ketoesters, BrF3 was able to convert the oxime group into a CF2 group and through a new type of rearrangement cause a shift of the carboxylate group to the nitrogen atom. The novel structure of the alpha,alpha-difluorocarbamate was also proven by 15N NMR as demonstrated for compounds 3, 8, 9, 12, 15, and 18. Another novel "double rearrangement" was observed during the formation of 19. Dynamic 19F NMR experiments indicate a high nitrogen inversion-rotation (NIR) barrier for these novel carbamates of about 12.5 kcal/mol. PMID- 11429821 TI - Asymmetric transformation of N-nitrosamines by inclusion crystallization with optically active hosts. AB - Several N-nitrosopiperidines with chirality solely due to a hindered rotation about the N-N bond were resolved to enantiomers by inclusion crystallization with optically active diols (TADDOLs). The absolute configuration of the guest nitrosamines was deduced from the X-ray crystal structures of the inclusion complexes. The enclathrated nitrosamines were liberated by a competitive complexation of the host diols with piperazine. The optical activity of the resolved nitrosamines is manifested by their CD spectra. A simple chirality rule was proposed for a rationalization of the observed Cotton effect sign corresponding to the n-pi* electronic transition. The optically active nitrosamines are configurationally labile compounds and gradually racemize in solution but they are indefinitely stable in the solid state. The first-order kinetics of the racemization in solution allowed us to assign the N-N rotation barriers by simple polarimetric measurements. PMID- 11429822 TI - Sequential nucleophilic substitution on halogenated triazines, pyrimidines, and purines: a novel approach to cyclic peptidomimetics. AB - A novel concept for the synthesis of macrocyclic peptidomimetics which incorporate heteroaromatic units is reported. The method involves sequential SNAr reactions of orthogonally protected amino groups of peptides and other linear oligomers on halogenated heterocycles such as 2,4,6-trichloro[1,3,5]triazine, 2,4,6-trichloropyrimidine, 4,6-dichloro-5-nitropyrimidine, and 2,6,8-trichloro-7 methylpurine. The scope of this novel solid-phase approach was systematically evaluated by means of the SPOT-synthesis methodology on planar cellulose membranes. Besides the question of the accessibility of different ring sizes and the compatibility with protecting groups of commonly used amino acids, the applicability of the technique toward different halogenated heteroaromatics and peptidomimetics was studied. It was found that the procedure is well suited to assemble a wide variety of cyclic peptidomimetics differing in both size (11- to 37-membered rings) and chemical nature of the assembled backbones. PMID- 11429823 TI - A novel reductive dimerization/oxidative dehydrogenation of aldimines mediated by lanthanoid metals. AB - A lanthanoid metal-mediated novel reductive dimerization/oxidative dehydrogenation of a variety of aldimines has been achieved. Aromatic aldimines (1) were dimerized in the presence of 0.5 mol of ytterbium metal (Yb) and 1 naphthaldehyde (1-NpCHO) to give the corresponding vicinal diimines (2) in good to high yields. Samarium metal (Sm) or samarium(II) diiodide (SmI2) gives unsatisfied yields under the same reaction conditions. As an oxidant, 1-NpCHO gives the best result. In addition, HMPA plays an important role as a cosolvent in this reaction. Reaction of various aldimines with Yb metal is discussed in detail. Moreover, it has been found that the electron-donating substitutents on the benzene ring promote the reaction and that the electron-withdrawing substitutents retard the reaction. The reaction of deuterated N benzylideneaniline with Yb metal confirmed that transformation of the hydride occurred from aldimines to the oxidant aldehyde. The mechanism of the reaction was discussed. PMID- 11429824 TI - Asymmetric epoxidation by chiral ketones derived from carbocyclic analogues of fructose. AB - A number of carbocyclic analogues of the fructose-derived ketone 1 have been prepared and investigated for asymmetric epoxidation. The studies show that the oxygen atom of the pyranose ring of 1 has an impact on the catalyst's activity and selectivity. Conformational, electronic, and steric effects are discussed. PMID- 11429825 TI - The synthesis of homoallylic amines utilizing a cuprate-based 1,2-metalate rearrangement. AB - Lithiation of the N-2,4,6-triisopropylbenzenesulfonyl-2-pyrroline (16) and treatment of the resulting cyclic vinyllithium reagent with R2CuCNLi2 produced an acyclic vinyl organometallic species that, when treated with an electrophile (H2O or RX), gave the homoallylic sulfonamides 18a-k in 37-93% yields and in > 95% diastereoselectivity. The deprotection of a representative homoallylic sulfonamide 18d was achieved in 83% yield by sonication in the presence of lithium wire and catalytic 4,4'-di-tert-butylbiphenyl (DBB). The efficacy of this general procedure for the production of homoallylic amine derivatives is demonstrated by the preparation of the diene amine 25, a key intermediate in the synthesis of a squalene synthetase inhibitor. PMID- 11429826 TI - Microbiological transformations. 47. A step toward a green chemistry preparation of enantiopure (S)-2-, -3-, and -4-pyridyloxirane via an epoxide hydrolase catalyzed kinetic resolution. AB - The biocatalyzed hydrolytic kinetic resolution of 2-, 3-, and 4-pyridyloxirane by the Aspergillus niger epoxide hydrolase (EH) has been explored. This was used to perform a gram scale preparation of these epoxides of (S) absolute configuration using a process performed at a concentration as high as 10 g/L (82 mM). All three epoxides have been obtained in a nearly enantiopure form (ee > 98%). Interestingly, it was shown that this biotransformation could be achieved using plain water instead of buffer solution, an important improvement as far as downstream processing of an eventual industrial process is concerned. Neither of these substrates could be obtained in reasonable enantiomeric purity and yield using the nowadays most efficient metal-based catalysts. PMID- 11429827 TI - Regioselective palladium-catalyzed synthesis of beta-arylated primary allylamine equivalents by an efficient Pd-N coordination. AB - A highly regioselective Heck arylation, utilizing aryl triflates and a palladium/dppf catalytic system, can be performed at the internal, beta-carbon of Boc- and phthalimido-protected allylamines, yielding arylated primary allylamine equivalents. The very high regioselectivity obtained with secondary Boc-protected allylamides is suggested to be caused by an efficient coordination between an anionic nitrogen and palladium. Single-mode microwave irradiation has been utilized to shorten the reaction times and, in the case of Boc-protected allylamides, to improve the yields of two electron-poor aryl triflates. PMID- 11429828 TI - Synthesis and functionalization of meso-aryl-substituted corroles. AB - The Rothemund condensation reaction of pyrrole and aldehydes is an extensively used route to meso-tetraarylporphyrins, but simple modifications of the reaction conditions allow the formation of different macrocycles other than the expected porphyrin. In the presence of an excess of pyrrole, this modified Rothemund approach leads to the synthesis of meso-triary-substituted corroles. This methodology allows the preparation of a wide range of substituted corroles starting from commercially available products. Higher yields have been obtained in the case of benzaldehydes bearing electron-withdrawing substituents, while the reaction fails in the presence of 2,6-disubstituted benzaldehydes. Although if not isolated, some experimental evidences indicate that the linear 5,10,15 triphenylbilane 4 is the precursor of the final corrole ring. Reaction of 5,10,15 triphenylcorrole 2 with an excess of NBS leads to the complete bromination of the macrocycle. Spectroscopic characterization seems to indicate the formation of the porphodimethene-like structure 5, where the macrocyclic aromatic conjugation is interrupted at the 10 position. Metalation of this compound with cobalt acetate and PPh3 affords the corresponding complex. The X-ray crystal structure of triphenylphosphine [2,3,7,8,12,13,17,18-octabromo-5,10,15-tris(4 nitrophenyl)corrolato]- cobalt(III) 8 confirms the ability of corrole ring to retain an almost planar conformation when fully substituted at the peripheral position. PMID- 11429829 TI - Synthesis of symmetrical polynitrohelicenes and their chiral recognition in the charge transfer complexation. AB - Derivatives of optically active 1,12-dimethylbenzo[c]phenanthrene-5,8 dicarboxylic acid can be nitrated regioselectively, giving symmetrically polyfunctionalized helicenes. The dicarboxylic acid or its dimethyl ester is dinitrated with fuming nitric acid in acetic acid at the 4,9-positions. When the reaction is conducted in fuming nitric acid, a 2,4,9,11-tetranitrohelicene is obtained. Analogously, 1,12-dimethylbenzo[c]phenanthrene-5,8-dinitrile gives 2,11 dinitro- or 4,9-dinitrohelicene depending on the conditions, and the former compound is converted to a 2,4,9,11-tetranitrohelicene. The tetranitrohelicenes form charge-transfer (CT) complexes with an electron-rich chiral diaminohelicene in solution. The studies on the chiral recognition reveal that the combinations of the same configuration of the helicenes form more stable complexes than that of the enantiomeric helicenes. PMID- 11429830 TI - Synthesis of trisubstituted furans from epoxypropargyl esters by sequential SmI2 promoted reduction-elimination and Pd(II)-catalyzed cycloisomerization. AB - A two-step one-pot synthesis of 2,3,5-trisubstituted furans 8 from 4,5-epoxyalk-2 ynyl esters 6 is described. The sequence is initiated by a SmI2-promoted reduction that takes advantage of the ability of the alkynyloxirane moiety present in 6 to accept electrons from SmI2. The resulting organosamarium species then eliminates an adjacent acetate or benzoate leaving group, leading to the formation of unstable 2,3,4-trien-1-ols 7. Without isolation, these are cycloisomerized to furans 8 by treatment with a catalytic amount of a Pd(II) complex and a proton source. The whole sequence takes place under mild reaction conditions. Some useful functional groups such as cyano and alpha,beta unsaturated esters are tolerated, but benzyl- and silyl-protected hydroxyl groups are deprotected to some extent. Starting materials can be easily assembled using reliable reactions from acetylene, an aldehyde or ketone, and a vinyl halide fragment. This offers the possibility of introducing branched substituents at C-5 of the furan ring. PMID- 11429831 TI - Reductive cross-coupling of 3-substituted delta 3-cephems with alkenyl halides in an Al/PbBr2/NiBr2(bpy) triplemetal redox system. Synthesis of 3-alkenyl-delta 3 cephems. AB - Synthesis of 3-alkenyl-delta 3-cephems was performed successfully by cross coupling 3-(trifluoromethylsulfonyloxy or chloro)-delta 3-cephem with alkenyl halides, e.g., vinyl bromide, trans-1-bromo-1-propene, and trans-beta bromostyrene in an Al/cat.PbBr2/cat.NiBr2(bpy)/NMP (or DMF) system. Reduction of 3-(trifluoromethylsulfonyloxy)-delta 3-cephem into norcephalosporin was also achieved by a similar reaction in the presence of 5 molar equiv of water. Scope and a plausible mechanism of the Al/Pb/Ni triplemetal-redox promoted reactions are discussed. PMID- 11429832 TI - Structural elucidation of a new cytotoxin isolated from mussels of the Adriatic sea. AB - A detailed analysis of the toxic composition in the hepatopancreas of mussels from northern Adriatic sea has been performed. Along with some polyether toxins of DSP (diarrhetic shellfish poisoning) type, such as yessotoxin and its analogues, which are responsible for a variety of human seafood poisonings throughout the world, we have now isolated a new type of toxin, the chlorosulfolipid 1, which is completely different in structure from the polyether DSP-toxins isolated so far. The structural determination of the new toxin, including its absolute stereochemistry, has been performed by extensive NMR analysis and molecular mechanics and dynamics calculations. PMID- 11429833 TI - A short stereoselective preparation of dienamides from cyclobutene compounds. Application in the synthesis of a new cyclohexene nucleoside. AB - A short stereoselective synthesis of N-acylamino-1,3-dienes was developed starting from the cyclobutene lactam 8, which was obtained from 2-hydroxypyridine by a photochemical electrocyclic reaction. The tert-butoxycarbonyl derivative 17 was prepared to facilitate nucleophilic attacks to the carbonyl group, and the subsequent thermal ring opening provided dienes 18-21. One of these (20) was used in the synthesis of the cyclohexene nucleoside 30. A Diels-Alder reaction between diene 20 and maleic anhydride provided the endo-cycloadduct 22a. Three additional steps yielded amine 26. Construction of the uracil moiety afforded intermediate 29. Cyclization and removal of the protecting groups occurred in one step in the presence of ammonia, giving the target molecule 30. Diene 20 also underwent [4 + 2] cycloaddition with methyl acrylate to provide predominantly the endo-product 23a, regioselectively. PMID- 11429834 TI - Reaction of vinyl epoxides with palladium-switchable bisnucleophiles: synthesis of carbocycles. AB - The selective activation of substrates I, potential bisnucleophiles, was achieved by using different palladium catalysts. The synthetic potential of this strategy has been demonstrated in the regiodivergent synthesis of carbocycles from substrates of type I, bearing malonate-type pronucleophiles and an alkenyl stannane, with vinyl epoxides. A selective palladium-catalyzed reaction of I with the vinyl epoxide gives rise to an allylic alcohol, which, after activation as a carbonate, led to the cyclization product by a second palladium-catalyzed reaction. The transmetalation process is favored with palladium-catalysts without phosphines or arsines as the ligands. On the other hand, the use of palladium complexes with PPh3 as the ligand inhibits the transmetalation pathway and promotes the nucleophilic attack of the malonate-type anions on the intermediate (eta 3-allyl)-palladium complexes. PMID- 11429835 TI - A simple method for the preparation of N-sulfonylsulfilimines from sulfides. AB - While excellent methods exist for the oxidation of sulfides to sulfoxides R1R2S- >R1R2SO, the azaversion of this atom transfer redox process, i.e., R1R2S- >R1R2S=N-SO2R3, has been less reliable. In sulfilimine synthesis, sulfoxide has been an inevitable byproduct in all cases to date, and the yields of sulfilimine have varied widely. A nearly ideal procedure for the sulfide to sulfonyl sulfilimine transformation is described. Almost quantitative yields are achieved from a diverse set of sulfides and a broad range of the readily available sulfonyl nitrenoid sources known as chloramine salts (R3SO2NClNa), essentially by simply stirring them together in acetonitrile. PMID- 11429836 TI - Photooxidative damage of guanine in DG and DNA by the radicals derived from the alpha cleavage of the electronically excited carbonyl products generated in the thermolysis of alkoxymethyl-substituted dioxetanes and the photolysis of alkoxyacetones. AB - On thermolysis of the methoxy (MeO-TMD), tert-butoxy (tBuO-TMD), and hydroxy (HO TMD) derivatives of 3,3,4,4-tetramethyl-1,2-dioxetane (TMD) in the presence of dG and calf-thymus DNA, the guanine is oxidized considerably more efficiently than the parent TMD. The same trend in the oxidative reactivity is observed for the photolysis of the corresponding oxy-substituted ketones versus acetone. The oxidative reactivity order in the dioxetane thermolysis, as well as in the ketone photolysis, parallels the ability of the excited ketones to release radicals (determined by spin trapping with DMPO and EPR spectroscopy) upon alpha cleavage (Norrish-type-I reaction). In the presence of molecular oxygen, the carbon centered radicals are scavenged to produce peroxyl radicals, which are proposed as the reactive species in the oxidation of the guanine in dG and calf-thymus DNA. PMID- 11429837 TI - A concise synthesis of fusaric acid and (S)-(+)-fusarinolic acid. PMID- 11429838 TI - A new protocol for Baylis-Hillman reactions: chirality transfer in a lithium phenylselenide induced tandem-michael-aldol-retro-michael reaction. PMID- 11429839 TI - An unusual oxidation of a benzylic methylene group by thionyl chloride: a synthesis of 1,3-dihydro-2-[2-(dimethyl-amino)ethyl]-1,3-dioxopyrrolo[3,4 c]acridine derivatives. PMID- 11429840 TI - Oxidative coupling of O-silyl and O-alkyl enethers: application of the novel annulation sequence to the synthesis of fluorinated naphthaldehydes and naphthyl ketones. PMID- 11429841 TI - Palladium-catalyzed asymmetric allylic substitution reactions using new chiral phosphinooxathiane ligands. PMID- 11429842 TI - A novel silylcopper catalyst for the reductive bromination of Hajos dione. Improved preparation of a CD synthon for the synthesis of vitamin D. PMID- 11429843 TI - Synthesis of a chiral aryl-ferrocenyl ligand, by intramolecular coupling to a biaryl-related lactone. PMID- 11429844 TI - Regioselective conversion of arylboronic acids to phenols and subsequent coupling to symmetrical diaryl ethers. PMID- 11429845 TI - Cycloisomerization of functionalized 1,5- and 1,6-dienes catalyzed by cationic palladium phenanthroline complexes. PMID- 11429846 TI - Total synthesis of indoles from Tricholoma species via Bartoli/heteroaryl radical methodologies. PMID- 11429847 TI - The first bovine beta 1,4-galactosyltransferase reaction with an acyclic acceptor substrate, 3-acetamido-1,2-propanediol, to yield a 3-O-beta-D-galactopyranosyl-sn glycerol skeleton. AB - [figure: see text] Reactivity of bovine beta 1,4-galactosyltransferase was examined for a series of acyclic acceptor substrates both in the presence and the absence of alpha-lactalbumin (alpha-La). It was found that this enzyme could utilize (R)-3-acetamido-1,2-propanediol (1) as an acceptor substrate regardless of the cofactor protein. The product was determined to be 1-O-beta-D galactopyranosyl-(R)-3-acetamido-1,2-propanediol (2). Glycerol without the acetamido group was inactive, indicating that this functional group plays a key role in the enzyme reaction. PMID- 11429848 TI - Pyrrolidyl polyamines: branched, chiral polyamine analogues that stabilize DNA duplexes and triplexes. AB - [figure: see text] Pyrrolidyl polyamines (III-VI) are conformationally restricted, chiral analogues of linear spermine elaborated by the addition of aminopropyl chains to yield branched diastereomers. It is demonstrated that in concentrations as low as 0.01 mM, these compounds remarkably stabilize DNA duplexes and triplexes through strong electrostatic interactions. The synthesized compounds are potential dendrons with a chiral pyrrolidine core, and such molecules may have potential as DNA delivery and transfection agents. PMID- 11429849 TI - Synthesis of the cyclohexan subunit of baconipyrones A and B from furan. AB - [figure: see text] The synthesis of the cyclohexan subunit of the siphonarlid metabolites baconipyrones A and B from furan is described. A key step included the alkylative ring opening of 7-oxanorbornenic sulfone 4 and oxidative desulfonylation of compound 8. PMID- 11429850 TI - Chiral auxiliaries for asymmetric radical cyclization reactions: application to the enantioselective synthesis of (+)-triptocallol. AB - [figure: see text] A series of epimeric 8-aryl menthyl derivatives 5a-d and 6a-l, prepared from the same chiral source (R)-pulegone, were employed as chiral auxiliaries in the asymmetric radical cyclization reactions of beta-keto esters mediated by Mn(OAc)3. Chiral precursors 8c and 8d provided the cyclization products 10c and 10d, respectively, as single isomers (dr > 99:1), whereas the cyclization of precursor 9k gave 13k with good stereoselectivity (dr = 24:1). Diastereomer 13e was employed as the key intermediate in the enantioselective synthesis of (+)-triptocallol in 90% ee. PMID- 11429851 TI - Facile entry to the tetracyclic 5-7-6-3 tigliane ring system. AB - [figure: see text] A tandem anionic 5-exo-dig cyclization/Claisen rearrangement sequence was used to effect a facile, "one-pot" conversion of an appropriately substituted 4-alkyn-1-ol to the tetracyclic carbon core structure of phorbol. The synthesis was conducted using readily available nonracemic starting materials to provide the target structure as a single enantiomer in high chemical yield. PMID- 11429852 TI - Palladium/imidazolium salt catalyzed coupling of aryl halides with hypervalent organostannates. AB - [figure: see text] A Pd(OAc)2/imidazolium chloride system has been used to mediate the catalytic cross-coupling of aryl halides with organostannanes. The imidazolium salt IPr.HCl (IPr = 1,3-bis(2,6-diisopropylphenyl) imidazol-2 ylidene) in combination with TBAF (nBu4NF) was found to be most effective for the cross-coupling of aryl bromides and electron-deficient aryl chlorides with aryl and vinyl stannanes. PMID- 11429853 TI - Stereoselective synthesis of dihydropyrone-containing marine natural products. Total synthesis and structural elucidation of (-)-membrenone-C. AB - [figure: see text] Three diastereomers of membrenone-C were separately prepared using a common two directional chain extending synthetic strategy. This has established the absolute and relative configuration of the natural product to be as shown in the foregoing graphic. Key steps in the synthesis of all the isomers are a stereoselective aldol coupling and reduction giving the C7-C9 stereocenters, a two direction chain extending double titanium aldol coupling, and the trifluoroacetic acid promoted double cyclization/dehydration giving the two dihydropyrone rings. PMID- 11429854 TI - Stereoselective synthesis of secondary organozinc reagents and their reaction with heteroatomic electrophiles. AB - [figure: see text] Various trisubstituted olefins were converted to configurationally stable diorganozinc compounds with high diastereoselectivity. Their reaction with various electrophiles centered on tin, sulfur, bromine, and phosphorus provided the desired substitution products with retention of configuration. Novel, functionalized organocopper reagents such as 4 and chiral diphosphine 5 have been prepared. PMID- 11429855 TI - Mesityllithium as a reagent for chemoselective halogen-lithium exchange reaction. AB - [figure: see text] Mesityllithium was found to be an excellent selective lithiating agent to prepare aryllithium compounds having alkoxycarbonyl groups. To extend our studies on chemoselective lithiation, an important precursor for the synthesis of camptothecin was prepared using a halogen-lithium exchange reaction followed by an intramolecular 1,2-addition. PMID- 11429856 TI - Novel synthesis of electroresponsive poly(thiophenylene) through a Michael-type addition. AB - [figure: see text] A novel poly(thiophenylene) having N,N-diphenyl-1,4 phenylenediamine (PDA) as a redox unit was synthesized through a Michael-type addition. This polymerization proceeded at room temperature without catalysts. The polymer obtained acted as a good electroresponsive material with moderate thermostability. PMID- 11429857 TI - Enantioselective syntheses of authentic sclerophytin A, sclerophytin B, and cladiell-11-ene-3,6,7-triol. AB - [figure: see text] Two distinctively different total syntheses of natural sclerophytin A in its revised structural formulation are reported. The first proceeds from (S)-carvone via a cladiellene triol and involves photoisomerization of the double bond. The second route makes use of (5S)-5-(d-menthyloxy)-2(5H) furanone, which is subjected to cycloaddition, Claisen ring expansion, and regiocontrolled dihydroxylation tactics. PMID- 11429858 TI - The copper-mediated cross-coupling of phenylboronic acids and N hydroxyphthalimide at room temperature: synthesis of aryloxyamines. AB - [figure: see text] A novel route to aryloxyamines via the copper-mediated cross coupling of N-hydroxyphthalimide and phenylboronic acids is reported. The reaction is mediated by selected copper(I) and (II) salts in the presence of pyridine and is tolerant of several functional groups on the phenylboronic acid. The phthallmide group is removed using hydrazine to afford the corresponding aryloxyamine. PMID- 11429859 TI - Efficient intramolecular asymmetric reductions of alpha-, beta-, and gamma-keto acids with diisopinocampheylborane. AB - [figure: see text] alpha-, beta-, and gamma-Keto acids are reduced with diisopinocampheylborane at room temperature to the corresponding hydroxy acids with predictable stereochemistry in very high ee. The gamma-hydroxy acids produced were conveniently cyclized to the corresponding lactones. This provides a simple synthesis of 4-hexanolide, a component of the pheromone secreted by the female dermestid beetle Trogoderma glabrum. PMID- 11429860 TI - Asymmetric synthesis of umuravumbolide. AB - [figure: see text] This first asymmetric synthesis of enantiopure desacetylumuravumbolide and umuravumbolide via asymmetric reduction, allylboration, and ring-closing metathesis confirms their revised structures and configurations. A convenient procedure to upgrade the enantiopurity of alpha,beta acetylenic alcohols is also described. PMID- 11429861 TI - Neisseria gonorrhoeae 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase does not catalyze the formation of the ribo analogue. AB - [figure: see text] Neisseria gonorrhoeae 3-deoxy-D-arabino-heptulosonate 7 phosphate (DAH 7-P) synthase catalyzes an aldol-type condensation between D erythrose 4-phosphate and phosphoenolpyruvate (PEP) to form 3-deoxy-D-arabino heptulosonate 7-phosphate and not 3-deoxy-D-riboheptulosonate 7-phosphate. Similar to the Escherichia coli enzyme, N. gonorrhoeae DAH 7-P synthase condenses D-arabinose 5-phosphate with PEP to give 3-deoxy-D-manno-octulosonate 8 phosphate. Therefore, the stereochemistry of the reaction catalyzed by N. gonorrhoeae DAH 7-P synthase at C1 of the phosphorylated monosaccharide is the same as that for the E. coli enzyme, namely, re face attack. PMID- 11429862 TI - Derailing the Wacker oxidation: development of a palladium-catalyzed amidation reaction. AB - [figure: see text] A conceptually new palladium-catalyzed amidation reaction is described for the synthesis of beta-amido ketones based on derailing the Wacker oxidation of enones. This reaction generates a new carbon-nitrogen bond via a palladium-catalyzed conjugate addition of a carbamate nucleophile to an enone. The regiocontrol, mild and neutral conditions, lack of preactivation of the nucleophile, and lack of reoxidation system for the catalyst are attractive features of this transformation. PMID- 11429863 TI - Highly stereoselective intramolecular Michael addition using alpha-sulfinyl vinyllithium as an unprecedented Michael donor. AB - [figure: see text] The first example of an asymmetric intramolecular Michael addition reaction using alpha-lithiated vinylic sulfoxide as a Michael donor is reported. Michael addition of the alpha-lithiated vinylic sulfoxide to (Z) enoates proceeds with high diastereoselectivity to give the adducts having a stereogenic center with (R)-configuration at the beta-position of the ester in the cyclopentene ring formation. The selectivity was reversed in the six-membered ring formation. On the other hand, the corresponding (E)-enoates provided Michael adducts with poor diastereoselectivity. PMID- 11429864 TI - Chelated aryllithium reagents: ring size and chelating group effects. AB - [figure: see text] Chelation and aggregation in phenyllithium reagents with potential 5-, 6-, and 7-ring chelating ether and amine ortho substituents have been examined utilizing variable-temperature 6Li and 13C NMR spectroscopy, 6Li and 15N isotope labeling, and the effects of solvent additives. Both ether and amine form strong 5-ring chelates; 6-ring ether chelates compete well with THF, but 6-ring amine chelates barely do, and 7-ring amine chelates do not. o Methoxymethylphenyllithium (4) forms an open dimer (9) and a pentacoordinate monomer with PMDTA (10). PMID- 11429865 TI - Entropy-controlled asymmetric synthesis. How differential activation entropy is induced in chiral tethered reactions. AB - [figure: see text] Kinetic measurements to determine effective molarities of intramolecular reactions using 2,4-pentanediol and related tethers showed that methyl groups on the tether accelerate the major diastereomeric process but decelerate the minor process. The efficient promotion of stereocontrol is suggested to be due to chiral perturbation of the reaction rate through the entropy term. The conformation of the encounter complex of the reagent and reactant moieties was deduced by stereochemical analysis of the intramolecular adducts. PMID- 11429866 TI - Highly enantioselective and regioselective biocatalytic azidolysis of aromatic epoxides. AB - [figure: see text] The halohydrin dehalogenase from Agrobacterium radiobacter AD1 catalyzed the highly enantioselective and beta-regioselective azidolysis of (substituted) styrene oxides. By means of kinetic resolutions the remaining epoxide and the formed azido alcohol could be obtained in very high ee. In a large scale conversion, the decrease in yield and selectivity due to the uncatalyzed chemical side reaction could be overcome by slow addition of azide. PMID- 11429867 TI - Photosensitized (electron-transfer) deconjugation of 1-arylcyclohexenes. AB - [figure: see text] A series of 1-arylcyclohexenes have been deconjugated to the corresponding 3-arylcyclohexenes via a photosensitized electron-transfer reaction. The introduction of substituents on the aryl group has provided insight into the underlying mechanism and has defined the scope and limitations of the reaction. PMID- 11429868 TI - The Cu(OTf)2- and Yb(OTf)3-catalyzed Claisen rearrangement of 2-alkoxycarbonyl substituted allyl vinyl ethers. AB - [figure: see text] The Cu(OTf)2- and Yb(OTf)3-catalyzed Claisen rearrangement of 2-alkoxycarbonyl-substituted allyl vinyl ethers has been developed. Reactivity and stereoselectivity strongly depend on the substrate structure. PMID- 11429869 TI - An azophenol-based chromogenic anion sensor. AB - [figure: see text] A new chromogenic azophenol-thiourea based anion sensor, 2, has been developed. This system allows for the selective colorimetric detection of F-, H2PO4-, and AcO-. Selectivity trends turned out to be dependent upon guest basicity and conformational complementarity between 2 and the guest. PMID- 11429870 TI - Exploring a benzyloxyaniline linker utilizing ceric ammonium nitrate (CAN) as a cleavage reagent: solid-phase synthesis of N-unsubstituted beta-lactams and secondary amides. AB - [figure: see text] A novel benzyloxyaniline linker that uses ceric ammonium nitrate (CAN) as a cleavage reagent is described. Its application in the solid phase synthesis of N-unsubstituted beta-lactams and secondary amides furnishes compounds in moderate to excellent yield (45-91%) and high purity (93-99%). PMID- 11429871 TI - Structure-activity relationships within a family of selectively cytotoxic macrolide natural products. AB - [figure: see text] We describe a semi-synthetic deglycosylated derivative of apoptolidin that retains considerable activity against the mitochondrial ATPase but has greatly reduced cellular cytotoxicity. We also demonstrate that a related antifungal natural product, cytovaricin, inhibits the same molecular target. Structural comparison of these macrolides provides insights into their conserved features that are presumably important for biological activity and identifies promising avenues at the interface of organic synthesis and biosynthesis for the generation of new selective cytotoxic agents. PMID- 11429872 TI - Intramolecular hydrosilylation and silicon-assisted cross-coupling: an efficient route to trisubstituted homoallylic alcohols. AB - [figure: see text] Alkylidenesilacyclopentanes (formed by intramolecular hydrosilylation of homopropargyl alcohols) are efficiently coupled with aryl or alkenyl halides in the presence of tetrabutylammonium fluoride and a palladium(0) catalyst. Yields of cross-coupling were generally high, and the reaction is compatible with a wide range of functional groups. The overall transformation achieves the conversion of homopropargyl alcohols to trisubstituted homoallylic alcohols in a highly stereoselective fashion. PMID- 11429873 TI - Reactions of cycloproparenes with metal carbenes. AB - [figure: see text] Benzocyclopropene and cyclopropa[b]naphthalene react with dichloro-bis(tricyclohexylphosphine)methylideneruthenium, incorporating the metallacarbene to form unstable 3-ruthenacyclopentenes, which decompose to give o xylylenes that can be trapped as Diels-Alder adducts by dimethyl acetylenedicarboxylate. In contrast, bis(eta 5 cyclopentadienyl)methylidenetitanium forms moderately stable 2- and 3 titanacyclopentene complexes. PMID- 11429874 TI - Regiocontrol of radical cyclization by Lewis acids. Efficient synthesis of optically active functionalized cyclopentanes and cyclohexanes. AB - [figure: see text] Treatment of alpha-alkylidenelactones 3a-d with Bu3SnH and a catalytic amount of Et3B effected a 5-exo radical cyclization preferentially to provide the corresponding 1 and 2 in a ratio of 70:30 to 100:0. Meanwhile, the reaction of 3a and 3b in the presence of Et2AlCl proceeded via a 6-endo cyclization pathway predominantly to afford 2a and 2b with 90% and 92% regioselectivity, respectively. PMID- 11429875 TI - Template-directed interference footprinting of protein-phosphate contacts in DNA. AB - [figure: see text] We have developed a method for interference footprinting of contacted phosphates in protein-DNA complexes. Template-directed enzymatic polymerization using a synthetic triphosphate analogue (alpha Me-dTTP) generates a product having a modified Internucleotide linkage, which perturbs protein phosphate contacts. We found that treatment of the methylphosphonodiester substituted extension product under nonaqueous conditions (MeO-/MeOH) led to the formation of a single cleavage product at each T residue but to two cleavage products when treated under the standard aqueous piperidine cleavage protocol. PMID- 11429876 TI - Kinetic dependence of the aqueous reaction of N-(hydroxymethyl)benzamide derivatives upon addition of electron-withdrawing groups. AB - [figure: see text] The rate constants for the hydronium ion, hydroxide, and water catalyzed breakdown of N-(hydroxymethyl)benzamide (1), 4-chloro-N (hydroxymethyl)benzamide (2), and 2,4-dichloro-N-(hydroxymethyl)benzamide (3) in H2O, at 25 degrees C, I = 1.0 (KCl), have been determined. The reactions of 1, 2, and 3 were found to be specific acid and specific base catalyzed with a first order dependence on hydronium and hydroxide ions. At higher hydroxide concentrations, the reactions were found to be pH independent for each compound studied. PMID- 11429877 TI - Diastereoselective epoxidation of oxazolidine-substituted alkenes by dimethyldioxirane and m-chloroperbenzoic acid: pi-facial control through hydrogen bonding by the urea functionality. AB - [figure: see text] A high diastereoselectivity (up to > 98:2) is found for the DMD and m-CPBA epoxidations of chiral oxazolidine-substituted olefins with a urea group. The selectivity is explained in terms of hydrogen bonding between the remote NH group of the urea functionality and the epoxidizing reagent. Methylation of the NH group prohibits hydrogen bonding, and a reversed selectivity is observed due to steric repulsion between the reagent and the urea functionality. PMID- 11429878 TI - Traceless solid-phase synthesis of substituted benzimidazoles via a base cleavable linker. AB - [figure: see text] A solid-phase route to substituted benzimidazoles has been developed using a modified base-labile linker strategy to release the final products in a traceless manner. This approach permits the synthesis of diverse compounds in moderate yields and high purity. PMID- 11429879 TI - Stereospecific reduction of phosphine oxides to phosphines by the use of a methylation reagent and lithium aluminum hydride. AB - [figure: see text] Various phosphine oxides are efficiently reduced by the use of a methylation reagent and lithium aluminum hydride. Optically active P-chirogenic phosphine oxides are also reduced with inversion of configuration at phosphorus atom by treatment with methyl triflate, followed by reaction with LiAlH4. PMID- 11429880 TI - High-yielding Staudinger ligation of a phosphinothioester and azide to form a peptide. AB - [figure: see text] The Staudinger ligation can be used to couple a peptide with a C-terminal phosphinothioester to another with an N-terminal alpha-azido group to form a single peptide that contains no residual atoms. Here diphenylphosphinomethanethiol thioesters are shown to give high isolated yields for this transformation. This finding provides precedent for a powerful and versatile new method for the total synthesis of proteins. PMID- 11429881 TI - Substituted alkyne synthesis under nonbasic conditions: copper carboxylate mediated, palladium-catalyzed thioalkyne-boronic acid cross-coupling. AB - [figure: see text] A new methodology for the synthesis of substituted alkynes is described. In the presence of copper(I) thiophene-2-carboxylate (CuTC) or copper (I) 3-methylsalicylate (CuMeSal), the palladium-catalyzed cross-coupling of thioalkyne derivatives with boronic acids affords functionalized alkynes in yields ranging from 39 to 91%. This coupling occurs efficiently under mild, nonbasic conditions with a wide variety of thioalkynes and boronic acids, providing a reaction complementary to the Sonogashira protocol. PMID- 11429882 TI - Improved method for rapid evaluation of chiral stationary phase libraries. AB - [figure: see text] An improved method for rapid LC/MS screening of chiral stationary phases based on the use of isotopically labeled enantiomers is reported. PMID- 11429883 TI - Cyanurate mimics of hydrogen-bonding patterns of nucleic bases: crystal structure of a 1:1 molecular complex of 9-ethyladenine and N-methylcyanuric acid. AB - [figure: see text] 9-Ethyladenine forms a unique molecular complex with N methylcyanuric acid consisting of homomeric and heteromeric hydrogen-bonding patterns. Also, the homomeric hydrogen bond pattern is different than that observed in its pure crystal structures. PMID- 11429884 TI - Facile solid-phase synthesis of sulfated tyrosine-containing peptides: total synthesis of human big gastrin-II and cholecystokinin (CCK)-39. AB - Chemical synthesis of tyrosine O-sulfated peptides is still a laborious task for peptide chemists because of the intrinsic acid-lability of the sulfate moiety. An efficient cleavage/deprotection procedure without loss of the sulfate is the critical difficulty remaining to be solved for fluoren-9-ylmethoxycarbonyl (Fmoc) based solid-phase synthesis of sulfated peptides. To overcome the difficulty, TFA mediated solvolysis rates of a tyrosine O-sulfate [Tyr(SO3H)] residue and two protecting groups, tBu for the hydroxyl group of Ser and 2,2,4,6,7 pentamethyldihydrobenzofuran-5-sulfonyl (Pbf) for the guanidino group of Arg, were examined in detail. The desulfation obeyed first-order kinetics with a large entropy (59.6 J.K-1.mol-1) and enthalpy (110.5 kJ.mol-1) of activation. These values substantiated that the desulfation rate of the rigidly solvated Tyr(SO3H) residue was strongly temperature-dependent. By contrast, the SN1-type deprotections were less temperature-dependent and proceeded smoothly in TFA of a high ionizing power. Based on the large rate difference between the desulfation and the SN1-type deprotections in cold TFA, an efficient deprotection protocol for the sulfated peptides was developed. Our synthetic strategy for Tyr(SO3H) containing peptides with this effective deprotection protocol is as follows: (i) a sulfated peptide chain is directly constructed on 2-chlorotrityl resin with Fmoc-based solid-phase chemistry using Fmoc-Tyr(SO3Na)-OH as a building block; (ii) the protected peptide-resin is treated with 90% aqueous TFA at 0 degree C for an appropriate period of time for the cleavage and deprotection. Human cholecystokinin (CCK)-12, mini gastrin-II (14 residues), and little gastrin-II (17 residues) were synthesized with this method in 26-38% yields without any difficulties. This method was further applied to the stepwise synthesis of human big gastrin-II (34 residues), CCK-33 and -39. Despite the prolonged acid treatment (15-18 h at 0 degree C), the ratios of the desulfated peptides were less than 15%, and the pure sulfated peptides were obtained in around 10% yields. PMID- 11429885 TI - 3-amino-1,2,4-benzotriazine 4-oxide: characterization of a new metabolite arising from bioreductive processing of the antitumor agent 3-amino-1,2,4-benzotriazine 1,4-dioxide (tirapazamine). AB - Tirapazamine (1) is a promising antitumor agent that selectively causes DNA damage in hypoxic tumor cells, following one-electron bioreductive activation. Surprisingly, after more than 10 years of study, the products arising from bioreductive metabolism of tirapazamine have not been completely characterized. The two previously characterized metabolites are 3-amino-1,2,4-benzotriazine 1 oxide (3) and 3-amino-1,2,4-benzotriazine (5). In this work, 3-amino-1,2,4 benzotriazine 4-oxide (4) is identified for the first time as a product resulting from one-electron activation of the antitumor agent tirapazamine by the enzymes xanthine/xanthine oxidase and NADPH:cytochrome P450 oxidoreductase. As part of this work, the novel N-oxide (4) was unambiguously synthesized and characterized using NMR spectroscopy, UV-vis spectroscopy, LC/MS, and X-ray crystallography. Under conditions where the parent drug tirapazamine is enzymatically activated, the metabolite 4 is produced but readily undergoes further reduction to the benzotriazine (5). Thus, under circumstances where extensive reductive metabolism occurs, the yield of the 4-oxide (4) decreases. In contrast, the isomeric two electron reduction product 3-amino-1,2,4-benzotriazine 1-oxide (3) does not readily undergo enzymatic reduction and, therefore, is found as a major bioreductive metabolite under all conditions. Finally, the ability of the 4-oxide metabolite (4) to participate in tirapazamine-mediated DNA damage is considered. PMID- 11429886 TI - Bone collagen cross-links: a convergent synthesis of (+)-deoxypyrrololine. AB - A convergent total synthesis of (+)-deoxypyrrololine (Dpl, 4), a putative cross link of bone collagen, is described starting from a commercially available L glutamic acid derivative, (4S)-5-(tert-butoxy)-4-[(tert-butoxycarbonyl)amino]-5- oxopentanoic acid (16). Condensation of aldehyde (S)-(-)-17 with nitro compound (S)-(-)-27, both of which were prepared from a common precursor (S)-16, gave the alpha-hydroxynitro compound 28, which upon acetylation afforded alpha acetoxynitro compound 14 in good yield. Subsequent condensation and cyclization of alpha-acetoxynitro compound 14 with benzyl isocyanoacetate (15) in the presence of DBU in THF gave the key pyrrole intermediate (S,S)-(-)-12 in 57% yield. N-Alkylation of pyrrole (S,S)-(-)-12 with iodide (S)-(-)-13 using t-BuOK in THF afforded the 2-benzyloxycarbonyl-1,3,4-substituted pyrrole derivative (-) 29 in 42% yield. Removal of the protective groups in (-)-29 followed by hydrogenolysis and decarboxylation afforded the cross-link (+)-Dpl (4) in good overall yield. The synthesis of an analogue (S)-(+)-24 and formation of a novel tetrahydroindole derivative (-)-31 are also described. PMID- 11429887 TI - A very short synthesis of steroids from 1,3-butadiene and benzocyclobutenes. AB - Lewis acid mediated addition of 1,8-bis(trimethylsilyl)octa-2,6-diene (BISTRO) 1 to succinic anhydride led to spirolactone 2 [(+/-)-6,9-divinyl-1 oxaspiro[4.4]nonan-2-one]. Methoxycarbonylation followed by stereoselective alkylation by various benzocyclobutenes afforded the substituted benzocyclobutene steroid precursors 5. Thermolysis of 5 gave rise to steroids (+/-)-6 with a trans anti-cis configuration in five steps and in a highly stereoselective manner. Modifications of the sequence allowed the preparation of steroids (+/-)-11 with trans-anti-trans configuration. PMID- 11429888 TI - [PtCl3(C2H4)]-[AmH]+ complexes containing chiral secondary amines: use as chiral derivatizing agents for the enantiodiscrimination of unsaturated compounds by 195Pt NMR spectroscopy and NMR stereochemical investigation. AB - Ionic complexes [PtCl3(C2H4)]-[AmH]+, containing chiral secondary amines, constitute a versatile class of chiral derivatizing agents (CDAs) for the enantiomeric purity determination of chiral unsaturated compounds via 195Pt NMR spectroscopy. The NMR conformational analysis allows us to search for the stereochemical basis of their enhanced versatility. PMID- 11429889 TI - First synthesis of totally orthogonal protected alpha-(trifluoromethyl)- and alpha-(difluoromethyl)arginines. AB - The first synthesis of a series of totally orthogonal protected racemic alpha (trifluoromethyl)- and alpha-(difluoromethyl)arginines is described. The key steps of the synthesis are the mild guanidinylation procedure and the selective hydrogenation of a CC triple bond in the presence of a Cbz-group. PMID- 11429890 TI - Amphidinolides T2, T3, and T4, new 19-membered macrolides from the dinoflagellate Amphidinium sp. and the biosynthesis of amphidinolide T1. AB - Three new 19-membered macrolides, amphidinolides T2 (2), T3 (3), and T4 (4), structurally related to amphidinolide T1 (1) have been isolated from two strains of marine dinoflagellates of the genus Amphidinium. The structures of 2-4 were elucidated on the basis of spectroscopic data. The absolute configurations at C 7, C-8, and C-10 of 1-4 were determined by comparison of NMR data of their C-1-C 12 segments with those of synthetic model compounds for the tetrahydrofuran portion. The biosynthetic origins of amphidinolide T1 (1) were investigated on the basis of 13C NMR data of a 13C enriched sample obtained by feeding experiments with [1-(13)C], [2-(13)C], and [1,2-(13)C2] sodium acetates and 13C labeled sodium bicarbonate in the cultures of the dinoflagellate. These incorporation patterns suggested that amphidinolide T1 (1) was generated from four successive polyketide chains, an isolated C1 unit formed from C-2 of acetates, and three unusual C2 units derived only from C-2 of acetates. Furthermore, it is noted that five oxygenated carbons of C-1, C-7, C-12, C-13, and C-18 were not derived from the C-1 carbonyl, but from the C-2 methyl of acetates. PMID- 11429891 TI - Enantioselective construction of cyclic quaternary centers: (-)-mesembrine. AB - The preparation of the crystalline amide 2 is reported. Conjugate addition to 2 proceeded with the expected high diastereocontrol to give 3. This set the stage for subsequent intramolecular alkylidene C-H insertion to give, after ozonolysis and aldol condensation, (-)-mesembrine 1. Amide 2 should be a useful chiron for the enantioselective construction of cyclic quaternary centers. PMID- 11429892 TI - Syntheses of pyrrolo- and indoloisoquinolinones by intramolecular cyclizations of 1-(2-arylethyl)-5-benzotriazolylpyrrolidin-2-ones and 3-benzotriazolyl-2-(2 arylethyl)-1-isoindolinones. AB - 1,5,6,10b-Tetrahydropyrrolo[2,1-alpha]isoquinolin-3(2H)-ones 17a,b, 17d,e, and 5,12b-dihydroisoindolo[1,2-alpha]isoquinolin-8(6H)-ones 22a-e were prepared by intramolecular cyclizations of 1-(2-arylethyl)-5-benzotriazolyl-pyrrolidin-2-ones 15a,b, 15d,e, and 3-benzotriazolyl-2-(2-arylethyl)-1-isoindolinones 20a-e, respectively, in the presence of titanium chloride. Products from chiral amines were obtained with stereoselectivities of > or = 94%. PMID- 11429893 TI - Inverted meso-aryl porphyrins with heteroatoms; characterization of thia, selena, and oxa N-confused porphyrins. AB - Synthesis and characterization of inverted porphyrins containing S, Se, and O are reported. A simple 3 + 1 MacDonald-type condensation using modified tripyrrane containing the N-confused ring and diols afforded various N-confused porphyrins 6a-f in 19-30% yield. The single-crystal X-ray structure of 6b shows a ruffled conformation with tilt angles of 21.11 degrees and 31.23 degrees for the N confused ring and the adjacent pyrrole ring III, respectively, revealing its severe nonplanarity. Significant changes in C alpha-C beta, C beta-C beta, and C alpha-X bond lengths are observed in 6b relative to free thiophene and pyrrole, suggesting the altered delocalization pathway in the modified N-confused porphyrins. The two molecules in the unit cell show a cyclophane-type noncovalent dimer with a face to face orientation of two N-confused pyrrole rings as a result of the presence of weak N-H...N and C-H...N intermolecular hydrogen bonds involving pyrrole-NH, the N atom of the N-confused ring, and the C atom of the pyrrole ring. A detailed 1H and 13C NMR study by 1D and 2D methods allowed assignments of all the peaks in the free base and protonated forms. NMR studies reveal the presence of three different tautomeric forms in solution for 6c in CDCl3 at low temperature. UV-visible studies reveal absorption band shifts upon heteroatom substitution, and the magnitudes of these shifts are dependent on the nature of the heteroatom. In all cases both monoprotonated and diprotonated species have been identified, and on addition of acid, the first proton goes to the outer N2 atom of the N-confused ring. PMID- 11429894 TI - Copper(I)-catalyzed [2 + 2] photocycloadditions with tethered linkers: synthesis of syn-photodimers of dicyclopentadienes. AB - Cu(I)-catalyzed intramolecular photocycloadditions of diesters made of endo dicyclopentadiene derivatives linked by the ester bonds with tethers are highly regio- and stereoselective and complete within hours, and the tethers can be easily cleaved afterward upon reduction with LiAlH4-Irradiation of the diesters afforded a 1:1 mixture of the heretofore unknown exo-cis-exo dimer, originating from the (R,S/S,R) diastereomer of the diester and the exo-trans-exo, deriving from the (R,R/S,S) diastereomer. The intermolecular photodimerization yielded, instead, only exo-trans-exo isomers and side products after irradiation for several days. The role of the tether's length and structure on the course of the photocycloadditions was investigated, and it was observed that short tethers introduce considerable strain in the products' framework. Adamantyl-containing tethers provided the shortest reaction times and highest yields. X-ray diffraction analysis of an exo-cis-exo stereoisomer containing adamantane in the tether exhibited an unusually close approach between H atoms on the methylene bridges and a long C-C distance in the cyclobutane ring. A rearrangement induced by X-ray irradiation was observed in this molecule. PMID- 11429895 TI - Ynolate chemistry. Reaction of a silylynolate with aziridines leading to gamma lactams. AB - A silylynolate, generated via the carbonylation of lithium silyldiazomethane, was reacted with N-tosyl aziridines to produce various five-membered lactams in good yields. The key step of this reaction involves the ring-opening ketenylation of aziridines by the silylynolate. The reaction proceeded in a highly stereoselective manner, and ketenylation took place at the less hindered carbon. When treated with aldehydes prior to protonation, the alpha-silylated lactam enolates gave alpha-vinylidene gamma-lactams. These reactions represent a unique path to the generation of and for controlling the reactivity of a rare class of reactive intermediates, namely, acyllithium derivatives and ynolates. PMID- 11429896 TI - A versatile palladium-mediated three-component reaction for the one-pot synthesis of stereodefined 3-arylidene-(or 3-alkenylidene-)tetrahydrofurans. AB - A one-pot reaction between equimolecular amounts of various propargyl alcohols, Michael acceptors, and unsaturated halides (or triflates) in the presence of a palladium(0) catalyst provides a simple and flexible entry into highly substituted 3-arylidene-(or 3-alkenylidene-)tetrahydrofurans. The efficiency of this palladium-mediated three-component reaction has been shown to be strongly influenced by the nature of the catalyst system, and in this regard, a palladium(0) catalyst generated in situ by reduction of PdCl2(PPh3)2 with n butyllithium has been found particularly effective. PMID- 11429897 TI - Palladium(II)-catalyzed oxidation of aldehydes and ketones. 1. Carbonylation of ketones with carbon monoxide catalyzed by palladium(II) chloride in methanol. AB - Unsubstituted or alkyl-substituted cyclic ketones react with PdCl2 in methanol under a CO atmosphere to give mainly acyclic diesters along with some acyclic chloro-substituted monoesters. The monosubstituted cyclic ketones, 2-hydroxy- and 2-methoxycyclohexanone, do not give ring cleavage but rather produce 2 (carbomethoxy)cyclohex-2-en-1-one. 13CO labeling experiments indicate one CO is inserted in forming the diester product so the second ester group must arise from the original ketone group. Two mechanisms are possible for the diester reaction. One involves initial Pd(II)-CO2CH3 insertion across the double bond of the enol form of the ketone while the second involves initial addition of Pd(II)-OCH3 followed by CO insertion into the new Pd(II)-carbon bond formed. Pd(II) elimination and acid-catalyzed ring cleavage produce the second methyl ester group in both routes. The chloro-substituted monoester is formed by initial Pd(II)-Cl insertion across the double bond followed by the acid-catalyzed ring cleavage. The 2-(carbomethoxy)cyclohex-2-en-1-one must result from elimination of water or methanol from the alpha-ketoester product formed by the initial methoxycarbonylation of the enol form of the ketone. As expected, the acyclic ketone, 2-decanone, formed methyl acetate and a mixture of methyl nonanoate and 1 chlorooctane as products. PMID- 11429898 TI - Aromatization of enamines promoted by a stoichiometric amount of palladium(II) salts: a novel method for the synthesis of aromatic amines. AB - Enamines (1a-r) prepared from cyclohexanones, cyclohexane-1,3-diones, or tetralones led to arylamines (2a-r) in one pot when treated with a stoichiometric amount of palladium salts [PdCl2-(MeCN)2] in acetonitrile in the presence of triethylamine at room temperature or at elevated temperature, in some cases for 5 min to 2 h. The initial electrophilic attack of palladium chloride on the beta carbon of the enamines led to a sigma-palladium species (8) which triggered a series of reactions (-->9-->10-->11-->12) destined for aromatization to give 2a-r in good yields. The intervention of such a sigma-palladium species has been attested by a trapping experiment. On the basis of this reaction mechanism, we have developed another new process capable of transforming acyclic compounds having 6-en-2-one frameworks (16, 23, 25) to arylamines (2s-u) when their enamines were treated under the similar conditions as above, featuring again the formation of sigma-palladium species such as 8 as the initial key intermediate. PMID- 11429899 TI - Selective hydrolysis of 2,4-diaminopyrimidine systems: a theoretical and experimental insight into an old rule. AB - Hydrolysis of the amino groups in condensed 2,4-diaminopyrimidine systems (1) has been used as a common method for the synthesis of oxo-substituted pyrimidines. In particular, the treatment with 6 M HCl usually yields exclusively the 2-amino-4 oxopyrimidine isomer (2). During our work, we found that the hydrolysis of the amino groups present in some condensed 2,4-diaminopyrimidine systems unexpectedly afforded exclusively the 4-amino-2-oxopyrimidine isomer (3). In this paper, we present the experimental work and ab initio calculations carried out to understand this discrepancy. As a part of such study, eight compounds containing a 2,4-diaminopyrimidine moiety were calculated in gas phase and in aqueous solution, and some acid hydrolyses were reexamined. Results showed that the presence of an electron-donating nitrogen linked to C6 of the 2,4 diaminopyrimidine ring changes the preferred hydrolysis site to yield the 4-amino 2-oxopyrimidine isomer. PMID- 11429900 TI - Palladium-catalyzed heteroannulation leading to heterocyclic structures with two heteroatoms: a highly regio- and stereoselective synthesis of (Z)-4-alkyl-2 alkyl(aryl)idene-3,4-dihydro-2H-1,4-benzoxazines and (Z)-3-alkyl(aryl)idene-4 tosyl-3,4-dihydro-2H-1,4-benzoxazines. AB - A highly convenient method has been developed for the synthesis of (Z)-4-alkyl-2 alkyl(aryl)idene-3,4-dihydro-2H-1,4-benzoxazines 9 and (Z)-3-alkyl(aryl)idene-4 tosyl-3,4-dihydro-2H-1,4-benzoxazines 34-38 through palladium-copper-catalyzed reactions. Aryl halides 7 reacted with 2-[N-alkyl(benzyl)-N-prop-2' ynyl]aminophenyl tosylate 6 in the presence of (PPh3)2PdCl2 (3 mol %), CuI(5 mol %) in triethylamine at room temperature to yield 2-[N-alkyl(benzyl)-N-(3-aryl prop-2'-ynyl)]-aminophenyl tosylates 8 in extremely good yields (72-96%). The latter could then be cyclized with KOH in ethanol-water to Z-9 in a highly regio- and stereoselective manner. Similarly, palladium-copper-catalyzed reaction of 2 (prop-2'-ynyloxy)aniline (21) with aryl iodides 7 led to 22-26 which after tosylation and cyclization with cuprous iodide in CH3CN in the presence of K2CO3 and Bu4-NBr led to the (Z)-3-alkyl(aryl)idene-4-tosyl 3,4-dihydro-2H-1,4 benzoxazines 34-38 in good overall yields. The Z-stereochemistry of the products was established from 1H NMR spectra, 3JCH values (between vinylic proton and methylenic carbon of the heterocyclic ring), NOE experiments, and X-ray analysis. The method was also found to be suitable for the synthesis of bis(benzoxazinylated) derivatives 17, 39, and 2-alkyl-3,4-dihydro-2H-1,4 benzoxazines 18. Our method for the synthesis of 3,4-dihydro-2H-1,4-benzoxazines is highly efficacious, using easily available starting materials under very mild conditions. Also the synthesis of some novel 5-substituted uracil derivatives 40 and 41 containing the benzoxazinyl moiety and of potential biological interest is being reported. PMID- 11429901 TI - Preparation and structural properties of 7,8-dioxa[6]helicenes and 7a,14c-dihydro 7,8-dioxa[6]helicenes. AB - The potentially chiral 7,8-dioxa[6]helicenes 1-1c have been prepared by oxidation of their precursors the 7a,14c-dihydro-7,8-dioxa[6]helicenes 3. The crystal structure determination of 3b cis-7a,14c-dihydro-3,12-dibromo-7,8 dioxa[6]helicene unambiguously confirms the cis configuration of the 7a,-14c hydrogens in compounds 3 as previously implied from NMR measurements and also shows that 3b crystallizes in a chiral conformation in the solid state. Selective deuteration of the sterically crowded 1,14 positions of 7,8-dioxa[6]helicene 1 influenced the crystal structure. The deuterium labeled compound D2-1 exhibits a disordered structure, whereas 1 had been found to crystallize in a complex structure which can be described as an analogous partly ordered modulated superstructure. When dehydrogenation of compound 3 to obtain compound 1 was attempted, harsh synthetic conditions gave the unexpected halogenated compounds 5 chloro-7,8-dioxa[6]helicene 1c and cis-7a,14c-dibromo-7,8-dioxa[6]helicene 3c. Compounds 1d and 3b were identified by solving their crystal structure. PMID- 11429902 TI - New, optically active phosphine oxazoline (JM-Phos) ligands: syntheses and applications in allylation reactions. AB - Three different syntheses of the phosphine oxazoline systems 1 are presented. Two of these approaches are divergent routes designed to involve an advanced intermediate that can be transformed into several different end-products. The third is a shorter route specifically designed to facilitate preparations of these systems on a larger scale using minimal functional group protection. Overall, eight different phosphine oxazolines were prepared. These were screened in several palladium-mediated allylation reactions. They proved to be most useful for asymmetric alkylation of 3-acetoxy-1,3-diphenylpropene and less suitable/effective for the more challenging substrates (a pentenyl derivative and a cyclohexenyl system). X-ray crystallographic analysis of the complex [(eta 3 PhCHCHCHPh)Pd(1a)][PF6] led to the conclusion that the origins of asymmetric induction in these systems might be indirectly attributed to interaction of the oxazoline-phenyl substituent with the palladium and with an allyl-phenyl substituent. Finally, data is presented for allylation of a silylenolate of an N acyl oxazolidinone; excellent enantioselectivities and yields were obtained. PMID- 11429903 TI - Single-component organic semiconductors based on novel radicals that exhibit electrochemical amphotericity: preparation, crystal structures, and solid-state properties of N,N'-dicyanopyrazinonaphthoquinodiiminides substituted with an N alkylpyridinium unit. AB - N,N'-Dicyanonaphthoquinodiimines fused with a pyrazine ring 1 were prepared from the corresponding quinones 4. The new acceptors 1 have a planar pi-system and undergo reversible two-stage 1e-reduction. Quaternization of the pyridyl substituent in 1d-f gave pyridinium derivatives 2d+, 2e+, and R-3+, respectively, which are stronger acceptors that undergo three-stage 1e-reduction. Upon electrochemical reduction of these cations, novel radicals 2d., 2e., and R-3. were generated and isolated as stable solids. The molecular geometries determined by X-ray analysis indicated that these radicals adopt a zwitterionic structure, in which the unpaired electron is located on the quinodiimine unit but not on the pyridyl group. These novel radicals undergo facile and reversible 1e-oxidation as well as two-stage 1e-reduction. The observed amphotericity endows the radicals with electrical conductivities (10(-5) to 10(-9) S cm-1), and these thus represent a new motif for single-component organic semiconductors. PMID- 11429904 TI - Molecular recognition studies on supramolecular systems. 32. Molecular recognition of dyes by organoselenium-bridged bis(beta-cyclodextrin)s. AB - A series of novel bis(beta-cyclodextrin)s tethered with organoselenium linkers, i.e., 6,6'-(o-phenylene-diseleno)-bridged bis(beta-cyclodextrin) (2), 6,6'-[2,2' diselenobis(benzoyloxy)]-bridged bis(beta-cyclodextrin) (3), and 6,6'-[2,2' diselenobis[2-(benzoylamino)ethylamino]]-bridged bis(beta-cyclodextrin) (4), were synthesized from beta-cyclodextrin (1). The inclusion complexation behavior of 1 4 with some dyes, such as 8-anilinonaphthalenesulfonate (ANS), Brilliant Green, Crystal Violet, Tropaeolin OO, Auramine O, and Methyl Orange, was investigated in aqueous phosphate buffer solution (pH 7.20) at 25 degrees C by UV-vis, fluorescence, and circular dichroism spectrometry, as well as fluorescence lifetime measurements. The complex stability constants (Ks) and Gibbs free energy changes (delta Go) for the stoichiometric 1:1 inclusion complexation of 1-4 with the dyes were obtained by the spectrophotometric or spectropolarimetric titrations. The bis(beta-cyclodextrin)s 2-4 showed much higher affinities toward these guest dyes than native beta-cyclodextrin 1 with fairly good molecular selectivities. The cooperative binding abilities of these bis(beta-cyclodextrin)s are discussed from the viewpoints of size/shape-fit interaction, induced-fit concept, and multiple recognition mechanism. PMID- 11429905 TI - Intramolecular photochemical dioxenone-alkene [2 + 2] cycloadditions as an approach to the bicyclo[2.1.1]hexane moiety of solanoeclepin A. AB - A synthesis of the bicyclo[2.1.1]hexane substructure of solanoeclepin A (1), the most active natural hatching agent of potato cyst nematodes, was approached via an intramolecular [2 + 2] photocycloaddition. Aldehyde 12 containing the dioxenone chromophore served as a useful starting material, allowing the synthesis of a variety of photocycloaddition substrates via Grignard addition or via a Nozaki-Hiyama-Kishi reaction. Photolysis of the unsubstituted alkene 14 led to the expected crossed cycloadduct bicyclo[2.1.1]hexane 15 according to the so called rule of five. However, several functionalized alkenes 18, 20, and 31 exhibited a complete reversal of cycloaddition regioselectivity, providing straight cycloadducts bicyclo[2.2.0]hexanes 21-26 and 4, respectively. Their structures were proved by a combination of extensive NMR measurements, X-ray analyses, and subsequent retro-aldol reactions. The latter de Mayo process allowed the formation of spiro-[3.5]nonane 35 and spiro[3.4]octane 36 as well as the cyclobutanes 37 and 38. Finally, the cyclization of the more rigid lactone precursor 28 occurred in high yield in the desired fashion with complete regio- and stereoselectivity to give 3 containing the core bicyclo[2.1.1]hexane skeleton of the natural product. PMID- 11429906 TI - A novel straightforward synthesis of enantiopure tetrahydroisoquinoline alkaloids. AB - A novel, direct,and high-yielding stereoselective method for enantiopure 1 substituted tetrahydroisoquinolines (THIQ) is described. The successful approach, which creates the stereocenter during the formation of the THIQ nucleus is based on (i) formation of chiral 2,3-substituted perhydro-1,3-benzoxazines derived from (-)-8-aminomenthol, (ii) diastereoselective intramolecular ring opening of the N,O-acetal moiety by an arylmetal generated from the substituent at the nitrogen atom in the perhydrobenzoxazine ring, and (iii) removal of the chiral auxiliary appendage. The starting perhydrobenzoxazines are easily prepared from (-)-8 aminomenthol and two different aldehydes, and the intramolecular opening is stereospecific, leading to a single stereoisomer. The method allows the preparation of a wide variety of enantiopure 1-substituted THIQ, with different substituents at C-1, by changing the nature of the starting aldehydes. PMID- 11429907 TI - Preparation and novel reduction reactions of vinamidinium salts. AB - Substituted acetic acids and formamides react in the presence of phosphorus oxychloride to yield the vinamidinium hexafluorophosphate salts 5a-d, 6a-d, and 7 in moderate to good unoptimized recrystallized yields (40-67%) as easily handled nonhygroscopic solids. The 1,3-differentially substituted vinamidinium salts 8 was prepared by amine exchange in 81% yield as are the cyclic diazapinium salts 9 and 10 in > 76% yield. The symmetrical 2-chlorovinamidinium 11 was prepared by displacement of 3 in 71% yield. The 2-chlorovinamidinium salts are cleanly reduced to the parent vinamidinium salts 12-16 using HI or PPh3/pTSA in up to 99% assay yield. PMID- 11429908 TI - Reaction of cyclohexanone benzylimines with ethylidenemalonate diesters. Diphenyl 2-ethylidenemalonate: a highly electrophilic synthetic equivalent of crotonic esters. AB - The diastereoselective Michael alkylation of alpha-substituted and alpha,alpha' disubstituted cyclohexanone benzylimines with ethylidenemalonate diesters was carried out for mechanistic and synthetic purposes. In the first case, an inverse regioselectivity occurred in comparison with what is generally observed since the Michael adducts resulted from alkylation of the non substituted enamine tautomer. With alpha,alpha'-disubstituted imines, in all cases, the stereochemistry of the major diastereomer was the one anticipated from a mechanism including a chairlike complex approach with a preferred exo position for the beta-methyl group of the ethylidenemalonic acid diesters. Furthermore, diphenyl 2-ethylidenemalonate 4 was found to be a highly electrophilic synthetic equivalent of crotonic esters. PMID- 11429909 TI - Regioselective enzymatic glycosylation of natural polyhydroxylated compounds: galactosylation and glucosylation of protopanaxatriol ginsenosides. AB - Ginsenoside Rg1 (1), the most representative Ginsenoside from Panax Ginseng C.A. Meyer belonging to the protopanaxatriol family, has been galactosylated by action of the beta-(1,4)-galactosyltransferase (GalT) from bovine colostrum, using UDP galactose as an activated sugar donor. The enzyme showed the well-known specificity for the formation of a beta-linkage with the C-4 OH of the glucose acceptor, but it was not able to discriminate between the two glucose moieties of 1, giving a mixture of mono- and digalactosylated derivatives. Other natural Rg1 analogues such as F1, Rh1, Re, as well as the synthetic derivative 6'-O-acetyl Rg1 have been also galactosylated, giving monolactosyl derivatives. GalT was also able to accept UDP-glucose as an activated sugar donor, giving rise to cellobiosyl derivatives of Rg1. PMID- 11429910 TI - Palladium-catalyzed addition of alcohol pronucleophiles to alkylidenecyclopropanes. AB - Palladium-catalyzed addition of alcohol pronucleophiles 1 to alkylidenecyclopropane derivatives 2 afforded the corresponding allylic ethers 3 in excellent to moderate yields. Catalyst optimization indicated the Pd(PPh3)4 P(o-tolyl)3 combination as the best system among all catalysts tested. Intramolecular reaction also proceeded smoothly. A plausible mechanism for this catalytic reaction was proposed. PMID- 11429911 TI - Intramolecular 1,3-dipolar cycloadditions of norbornadiene-tethered nitrile oxides. AB - Efficient routes to the synthesis of norbornadiene-tethered nitrile oxides have been developed, and their intramolecular 1,3-dipolar cycloadditions were studied. The cycloadditions occurred in good yields for a variety of substrates and were found to be highly regio- and stereoselective, giving single regio- and stereoisomers in most cases. PMID- 11429912 TI - Photochemical generation and matrix-isolation detection of dimethylvinylidene. AB - We report the spectroscopic characterization of dimethylvinylidene, (CH3)2C=C:, generated within an argon matrix at 14 K from a bisperoxyester precursor. The carbene was identified by comparison of the experimental IR spectrum with vibrational frequencies computed at the B3LYP/6-31G(d) level. Chemical trapping of the carbene within a 9% CO/Ar matrix to form dimethylpropadienone supports this analysis. Additional products produced during photolysis were identified by comparison to the appropriate computed vibrational frequencies. The potential energy surface of dimethylvinylidene and its intramolecular rearrangement products, 2-butyne and methylcyclopropene, were also investigated computationally at the B3LYP/6-31G(d) level. A spin-state analysis of this carbene using a variety of computational methods (CCSD(T), B3LYP, MP2) indicates the singlet state is more stable than the triplet by approximately 45 kcal mol-1. We anticipate the bisperoxyester precursor used here will be a convenient and general way for initiating future studies of alkylvinylidenes under matrix isolation conditions. PMID- 11429913 TI - Investigations in the transition metal catalyzed aziridination of olefins, amination, and other insertion reactions with Bromamine-T as the source of nitrene. AB - Investigations into the transition metal catalyzed aziridination of olefins with Bromamine-T as a new source of nitrene is presented in this account. Comparison of Chloramine-T and Bromamine-T in this reaction indicates that the latter is superior as the source of nitrene. Systematic study with several transition metal based catalysts suggests that Cu-halides are the best catalysts. A first report of aziridination under microwave and ultrasound irradiation conditions is also presented. Copper-catalyzed aziridination of methyl cinnamate with Bromamine-T did not proceed at ambient temperature but was effected smoothly under ultrasound irradiation to furnish trans-aziridine selectively, while under microwave irradiation, a mixture of cis and trans isomers, was obtained. It has been demonstrated that aziridination of olefins proceeds smoothly with inexpensive bleaching powder. Preliminary results of Rh-catalyzed benzylic insertion reactions with Bromamine-T are included in this account. PMID- 11429914 TI - Oxovanadium(V)-induced oxidative ligand coupling of aryltrimethylzincates prepared from bromoarenes and dilithium tetramethylzincate. PMID- 11429915 TI - Asymmetric borane reduction of prochiral ketones by polymer-supported chiral sulfonamides. PMID- 11429916 TI - Rearrangement of alpha-amino cyclopropanone hydrate: a novel route to labeled amino acids. PMID- 11429918 TI - Copper-catalyzed cyclization of N-allylhalodifluoroacetamides: an efficient synthesis of alpha,alpha-difluorinated gamma-lactams. PMID- 11429917 TI - Total synthesis of (-)-heliannuol E. PMID- 11429919 TI - Facile and efficient transformation of xanthates into thiocarbonates by anodic oxidation. PMID- 11429920 TI - Enzyme-assisted enantioselective synthesis of natural (-)-beta-necrodol and its enantiomer. PMID- 11429921 TI - Novel synthesis of [33P]-(2-chloroethyl)phosphonic acid. PMID- 11429922 TI - Reduction and coupling reactions of carbonyl compounds using samarium metal in aqueous media. PMID- 11429923 TI - A novel and selective approach to enantiomerically pure bicyclic-trans-lactams via a titanium enolate of a thiopyridyl ester. PMID- 11429924 TI - Synthesis of bicyclic ortho esters by epoxy ester rearrangements and study of their ring-opening reactions. PMID- 11429925 TI - Application of NAD(P)H model Hantzsch 1,4-dihydropyridine as a mild reducing agent in preparation of cyclo compounds. PMID- 11429926 TI - Synthesis of (S)-alpha-cyclopropyl-4-phosphonophenylglycine. PMID- 11429927 TI - Electrophilic nitration of aromatics in ionic liquid solvents. AB - Potential utility of a series of 1-ethyl-3-methylimidazolium salts [emim][X] with X = OTf-, CF3COO-, and NO3- as well as [HNEtPri2][CF3COO] (protonated Hunig's base) ionic liquids were explored as solvent for electrophilic nitration of aromatics using a variety of nitrating systems, namely NH4NO3/TFAA, isoamyl nitrate/BF3.Et2O, isoamyl nitrate/TfOH, Cu(NO3)/TFAA, and AgNO3/Tf2O. Among these, NH4NO3/TFAA (with [emim][CF3COO], [emim][NO3]) and isoamyl nitrate/BF3.Et2O, isoamyl nitrate/TfOH (with [emim][OTf]) provided the best overall systems both in terms of nitration efficiency and recycling/reuse of the ionic liquids. For [NO2][BF4] nitration, the commonly used ionic liquids [emim][AlCl4] and [emim][Al2Cl7] are unsuitable, as counterion exchange and arene nitration compete. [Emim][BF4] is ring nitrated with [NO2][BF4] producing [NO2 emim][BF4] salt, which is of limited utility due to its increased viscosity. Nitration in ionic liquids is surveyed using a host of aromatic substrates with varied reactivities. The preparative scope of the ionic liquids was also extended. Counterion dependency of the NMR spectra of the [emim][X] liquids can be used to gauge counterion exchange (metathesis) during nitration. Ionic liquid nitration is a useful alternative to classical nitration routes due to easier product isolation and recovery of the ionic liquid solvent, and because it avoids problems associated with neutralization of large quantities of strong acid. PMID- 11429928 TI - Alkylation of amino acids and glutathione in water by o-quinone methide. Reactivity and selectivity. AB - o-Quinone methide (1) has been produced in water both thermally and photochemically from (2-hydroxybenzyl)trimethylammonium iodide (2). Michael addition reactions of 1 to various amines, and sulfides, including amino acids and glutathione have been carried out, obtaining alkylated adducts (3-16) in fairly good to quantitative yields. The reaction rate and selectivity of 1 toward nitrogen and sulfur nucleophiles, in competition with the hydration reaction, have been investigated at different pH by laser flash photolysis technique. The observed reactivity spans 7 orders of magnitude on passing from water (kNu = 5.8 M-1 s-1) to the most reactive nucleophile (2.8 x 10(8) M-1 s-1, 2-mercaptoethanol under alkaline conditions). These are the first direct reaction rate measurements of nucleophilic addition to the parent o-quinone methide (1). Competition experiments provided strong kinetic support to the involvement of free 1 as an intermediate in both thermal and photochemical reactions. Furthermore, several alkylation adducts regenerate 1 either by heating (9, 10, 13, and 14) or by irradiation (9, 11-13, 16). Such a thermal and photochemical reversibility of the alkylation process opens a new perspective for the use and application of such adducts as o-QM molecular carriers. PMID- 11429929 TI - Synthesis of 2,3-disubstituted pyrroles and pyridines from 3-halo-1-azaallylic anions. AB - A new synthesis of 2,3-disubstituted pyrroles and pyridines is described. The reaction of 3-halo-1-azaallylic carbanions, regiospecifically generated from alpha-halogenated ketimines, with omega-iodoazides led to the regiospecific formation of omega-azido-alpha-haloketimines. Treatment of these functionalized imines with tin(II) chloride afforded halogenated five- and six-membered cyclic imines, which were transformed under mild conditions into 2,3-disubstituted pyrroles and pyridines. The stereoselective reduction of 2,3-dialkyl-3-chloro-1 pyrrolines to afford cis-2,3-dialkyl-3-chloropyrrolidines is also reported. PMID- 11429930 TI - Hypervalent iodine(III)-induced intramolecular cyclization of alpha-(aryl)alkyl beta-dicarbonyl compounds: a convenient synthesis of benzannulated and spirobenzannulated compounds. AB - A novel hypervalent iodine(III)-induced direct intramolecular cyclization of alpha-(aryl)alkyl-beta-dicarbonyl compounds has been described. Both meta- and para-substituted phenol ether derivatives containing acyclic or cyclic 1,3 dicarbonyl moieties at the side chain undergo this reaction in a facile manner. The reactions afford benzannulated and spirobenzannulated compounds that are of biological importance. The reaction is found to be general, mild, and high yielding. The mechanism of the reaction has been shown to involve a cation radical intermediate. PMID- 11429931 TI - Trapping of 1,8-biradical intermediates by molecular oxygen in photocycloaddition of naphthyl-N-(naphthylcarbonyl)carboxamides; formation of novel 1,8-epidioxides and evidence of stepwise aromatic cycloaddition. AB - The photocycloaddition reaction of naphthyl-N-(naphthylcarbonyl)carboxamides (1) was examined under argon and oxygen atmospheres. In addition to the [2 + 2] and [4 + 4] cycloadducts, 3 and 4, respectively, novel 1,8-epidioxides (5) were formed under oxygen atmosphere. The transient absorption at lambda max of 360 nm with the lifetime of 360 ns was observed by laser flash photolysis of 1c and was interpreted as the absorption of biradical intermediate 2. On the basis of the anti stereochemistry of 5, which was different from that of the major [4 + 4] cycloadducts, syn-4, it was deduced that equilibrium between biradical intermediates syn-2 and anti-2 would exist. Retro [2 + 2] cycloaddition of 3 was responsible for the efficient trapping of the biradical intermediate with molecular oxygen. The photocycloaddition of the anthryl derivatives, 9-anthryl-N (methylethyl)-N-(naphthylcarbonyl)carboxamides (7), afforded the [4 + 4] cycloadducts (8) exclusively in a quantitative yield even under oxygen atmosphere. The absence of trapping with molecular oxygen was interpreted to be due to the lack of retro [4 + 4] cycloaddition of 8. PMID- 11429932 TI - Alkyne-based, highly stereo- and regioselective synthesis of stereodefined functionalized vinyl tellurides. AB - (Z)-beta-Aryltellurovinylphosphonates and (Z)-beta-aryltellurovinyl sulfones were synthesized via the highly stereoselective anti-hydrotelluration of 1 alkynylphosphonates and 1-alkynyl sulfones. The configurations of these compounds were characterized via 1H NMR spectra or NOESY experiments and by X-ray diffraction analysis; (E)-beta-aryltellurovinyl sulfones were obtained with the reaction of sodium aryltellurate with (E)-2-iodovinyl sulfones to confirm the stereochemistry of the above anti-hydrotelluration. When the tandem reaction of alkynes with diaryl ditellurides and sodium arylsulfinates was carried out in AcOH/H2O (4/1), the corresponding (E)-beta-aryltellurovinyl sulfones were obtained in one step in good yields. This reaction is highly regio- and stereoselective and proceeds by using arylsulfinate as the sulfonyl radical precursor and diaryl ditellurides as free radical acceptors. (E)-1-Iodo-2 aryltelluroalkenes can be obtained by the anti-addition of ArTeI with terminal alkynes in THF. The stereochemistry of compound 17b was also determined by X-ray diffraction analysis. PMID- 11429933 TI - Asymmetric total synthesis of halicholactone. AB - The asymmetric total synthesis of the marine metabolite, halicholactone 1, is described. The bisallylic triol 6 with three chiral centers at C8, C12, and C15 was constructed by [2,3]-sigmatropic rearrangement of the sulfoxide 18, which was prepared stereoselectively using the chirality of (diene)Fe(CO)3 complexes. Introduction of the trans-substituted cyclopropane subunit into 21 was successfully achieved using the modified regio- and stereoselective Simmons-Smith reaction. The use of RCM (ring-closing metathesis) methodology (4-->35) was pivotal for the formation of a nine-membered unsaturated lactone fragment of halicholactone 1. As this approach is flexible and stereoselective, other oxylipins could be synthesized by the protocol described herein. PMID- 11429934 TI - Synthesis of nickel(II) triazolenaphthalocyanine and related macrocycles. AB - The synthesis of novel monbenzotriazolephthalocyanines (3-6), low-symmetry phthalocyanine analogues, is described for the first time using a novel and simple stepwise strategy which involves the use of substituted naphthalodinitriles 8-11 and appropriately substituted three-unit compounds such as 7. Belonging to the same family, compound 1, which we have called triazolenaphthalocyanine, has been prepared by a statistical method from compound 2 and 3,5-diamino-1,2,4-triazole. Compound 1 shows a bathochromic shift of the Q band in UV-vis spectrum due to the fusing of benzene rings to the triazolephthalocyanine core. PMID- 11429935 TI - A "green" route to perylene dyes: direct coupling reactions of 1,8-naphthalimide and related compounds under mild conditions using a "new" base complex reagent, t BuOK/DBN. AB - The direct coupling reactions of 1,8-naphthalimide compounds efficiently occurred at 130 or 170 degrees C without the intervention of the leuco form dyes in the presence of base complex reagent, t-BuOK/1,5-diazabicyclo[4.3.0]non-5-ene (DBN), to give the corresponding perylene dyes in good yields with > 95% purities. A possible mechanistic speculation for these oxidative coupling reactions is briefly discussed. PMID- 11429936 TI - Rearrangements of 3-aryl-substituted cyclopropenyl anions and the gas-phase acidity of 3-(4-methylphenyl)cyclopropene. AB - 3-(4-Methylphenyl)-3-trimethylsilylcyclopropene and 3-(4-trifluoromethylphenyl)-3 trimethylsilylcyclopropene react with fluoride ion in the gas phase to afford 6 substituted 3-indenyl anions via a spontaneous rearrangement of their corresponding cyclopropenyl anions. These isomerizations led us to reinvestigate the reported gas-phase generation of 1,2,3-triphenylcyclopropenyl anion, and contrary to the previous study, a similar rearrangement to 1,2-diphenyl-1-indenyl anion is observed. Despite the instability of 3-aryl-3-cyclopropenyl anions, we were able to measure the acidity of 3-(4-methylphenyl)cyclopropene at the allylic position (delta H(o)acid = 398.6 +/- 1.4 kcal/mol) by the DePuy kinetic method. Ab initio calculations on the structures and energies of mono- and triaryl substituted cyclopropenyl anions also are presented. PMID- 11429937 TI - In vivo early plaque formation on pure titanium and ceramic abutments: a comparative microbiological and SEM analysis. AB - A lack of information exists about the influence of different implant abutment materials on bacterial colonization and its role in the development of perimplantar infections. In order to study these aspects, removable acrylic devices, harboring samples of titanium and novel ceramic abutments (Nobel Biocare) were adapted to the molar-premolar region in 2 mandibular quadrants of 4 volunteers. Samples of each material were collected at 6 and 24 h, 7 and 14 days. Samples were observed by scanning electron microscopy and bacterial counts were made by means of ATP detection and direct plate count. The electron micrographs demonstrated that the bacteria colonization was already present after 6 h of presence in the oral cavity. After 24 h, both the materials were covered by several layers of bacterial cells. No differences in microbial colonization were observed between titanium and ceramic samples. The microbiological analysis confirmed the presence of relevant amounts of microbial cells on the tested samples. The maximum of colonization was achieved after 24 h in the oral cavity and the bacterial counts remained constant over the 14 day period. No significant differences were observed between the two materials analyzed in this study. In addition, ATP-bioluminescence technology was demonstrated to be a suitable system to evaluate bacterial colonization in the oral cavity. PMID- 11429938 TI - C-telopeptide pyridinoline cross-links (ICTP) and periodontal pathogens associated with endosseous oral implants. AB - Detection of periodontal or peri-implant sites exhibiting progressing disease or those at risk of deterioration has proven difficult. Pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP), a marker specific for bone degradation found in gingival crevicular fluid (GCF), has been associated with both bone and attachment loss in periodontitis and may be useful for predicting disease activity. The aim of this cross-sectional study was to examine the relationship between ICTP levels and subgingival species around implants and teeth from 20 partially and 2 fully edentulous patients. GCF and plaque samples were collected from the mesiobuccal site of each implant and tooth. Radioimmunoassay techniques were utilized to determine GCF ICTP levels. Plaque samples were analyzed utilizing checkerboard DNA-DNA hybridization. Traditional clinical parameters were assessed. Seventy-one implants and 370 teeth from 22 subjects were examined. ICTP levels and subgingival plaque composition were not significantly different between implants and teeth. Implant sites colonized by Prevotella intermedia, Capnocytophaga gingivalis, Fusobacterium nucleatum ss vincentii, and Streptococcus gordonii exhibited odds ratios of 12.4, 9.3, 8.1, and 6.7, respectively of detecting ICTP. These results suggest a relationship between elevated ICTP levels at implant sites and some species associated with disease progression. Longitudinal studies are necessary to determine whether elevated ICTP levels may predict the development of peri-implant bone loss. PMID- 11429939 TI - Mandibular overdentures supported by two Branemark, IMZ or ITI implants. A prospective comparative preliminary study: one-year results. AB - The aim of this prospective comparative study was to evaluate the condition of the peri-implant tissues of three different implant systems supporting a mandibular overdenture. Ninety edentulous patients (Cawood class V-VI) participated in this study. After randomization, 30 patients were treated with 2 Branemark implants, 30 patients with 2 IMZ implants and 30 patients with 2 ITI implants. The implants were inserted in the canine region of the mandible. After 3 months overdentures were fabricated supported by a round bar and clip attachment. A standardized clinical and radiographic evaluation was performed 0,6 and 12 months after insertion of the denture. The intraoral radiographs were made, using the long-cone technique with an aiming device. Two implants were lost (1 Branemark, 1 IMZ) during the healing period. None of the patients showed any sensory change in lip or chin region. The pocket depth in the Branemark group decreased significantly whereas the mucosa recession increased significantly in both the Branemark as well as in the IMZ group. After 12 months, there was significantly less bone loss in the ITI group. From our study it was concluded that 2 (Branemark, IMZ or ITI) implants placed in the interforaminal region connected with a bar supply a proper base for the support of a mandibular overdenture in the (Cawood V-VI) edentulous patient. The ITI implant appears to be the implant of choice for mandibular overdenture therapy, because only one operation is required for a comparable result. PMID- 11429940 TI - The effect of hyperbaric oxygen treatment on bone tissue reactions to c.p. titanium implants placed in free autogenous bone grafts. A histomorphometric study in the rabbit mandible. AB - This study aimed to investigate the effect of hyperbaric oxygen (HBO) therapy on the tissue reactions to commercially pure (c.p.) titanium implants placed in free autogenous bone graft by a 1-stage procedure. Eighty c.p. titanium implants were placed in the bone grafted from iliac crest to bilateral mandible of 40 Japanese white rabbits without tapping. Twenty rabbits underwent daily a HBO treatment for 60 min under 2.4 ATA during 20 consecutive days and the other untreated 20 rabbits served as controls. The implants with surrounding bone tissue were retrieved 20, 30, 60, 90 and 120 days after surgery, fixated, dehydrated and embedded in resin. About 20 microns thick ground sections were prepared prior to microscopical observations. The bone area and the bone-to-implant contact inside the threads were calculated separately in the grafted bone and in the host bone for each implant. After 30 and up to 120 days, the HBO treated group showed more bone-to-implant contacts in the grafted bone as compared to the non-HBO treated group. In the host bone there were no differences observed between HBO and non HBO treated groups. This study indicated that HBO treatment was beneficial for the tissue incorporation of c.p. titanium implants when placed immediately in free autogenous bone grafts. PMID- 11429941 TI - Use of 5-mm-diameter implants: Periotest values related to a clinical and radiographic evaluation. AB - A modified design of the original Branemark implant consisting of a cp. Titanium 5.0-mm-diameter self-tapping implant threaded up to the marginal platform has been proposed for specific indications. From February 1992 to November 1995, a total of 185 machined screw implants (Nobel Biocare, Gothenburg, Sweden) were installed in 45 patients to withstand 58 prostheses. Of these, 91 were 3.75-mm diameter and 94 were 5.0-mm wide. Most of the implants were placed in type B and C bone quantity and type 2 and 3 bone quality. A retrospective evaluation with regard to indications, marginal bone remodelling, Periotest values (PTv) and survival rate is presented. PTv and radiographic measurements were made at abutment connection and repeated 3, 6 and 12 months later and thereafter every year. The follow-up ranged from 16 to 55 months (mean 32.9 months) post-loading. Three patients with 8 5.0-mm implants dropped-out of the study at different stages. Out of the wide implants, 1 was expelled during the healing period; 3 were found mobile at the abutment connection; 1 lost its osseointegration suddenly after 2 years of function; 4 belonging to 1 patient did not meet the success criteria due to continuous marginal bone loss. The cumulative success rate of 5.0-mm implants (CSR) after 1 year of function was 97.2% for upper jaws and 88.4% in mandibles, whereas the CSR in maxilla after 48 months was 97.2% and 83.4% in mandibles. The obtained PTv from 5.0-mm-wide fixtures in maxilla and mandibles were respectively 1.1 and 0.6 units lower than those obtained PTv for 3.75-mm-diameter implants in the same patients. The hypothesis that there are differences in the damping capacity of the bone surrounding a 5.0-mm-wide implant compared to the 3.75-mm-diameter implant is supported by the PTv results. PMID- 11429942 TI - The influence of bone mechanical properties and implant fixation upon bone loading around oral implants. AB - Finite element models were created to study the stress and strain distribution around a solitary Branemark implant. The influence of a number of clinically relevant parameters was examined: bone-implant interface (fixed bond versus frictionless free contact), bone elastic properties, unicortical versus bicortical implant fixation and the presence of a lamina dura. Bone loading patterns in the vicinity of the implant seem to be very sensitive to these parameters. Hence they should be integrated correctly in numerical models of in vivo behaviour of oral implants. This necessitates the creation of patient dependent finite element models. PMID- 11429943 TI - Alveolar bone formation at dental implant dehiscence defects following guided bone regeneration and xenogeneic freeze-dried demineralized bone matrix. AB - The present study evaluated rate and extent of alveolar bone formation in dental implant dehiscence defects following guided bone regeneration (GBR) and implantation of xenogeneic freeze-dried demineralized bone matrix (xDBM). A total of 16 titanium plasma-sprayed (TPS) and 16 hydroxyapatite-coated (HA) titanium cylinder implants were inserted in 4 mongrel dogs following extraction of the mandibular premolar teeth. Four implant sites per jaw quadrant (2 TPS and 2 HA implant sites) were prepared into extraction sockets in each dog. Buccal alveolar bone was removed to create 3 x 5 mm dehiscence defects. Two jaw quadrants in separate animals received GBR, GBR + xDBM, xDBM (control), or gingival flap surgery alone (GFS; control). Thus, four conditions were available for each implant type (TPS or HA): GBR, GBR + xDBM; xDBM and GFS. The animals received fluorescent bone labels to allow observations of rate and extent of bone formation. Animals were sacrificed at 12 weeks postsurgery and block sections were harvested for histologic analysis. There were no apparent histologic differences between TPS and HA implant defects. GBR and GBR + xDBM resulted in almost complete bone closure of the dental implant dehiscence defect. Rate of bone formation appeared higher following GBR alone. Extent of bone formation appeared somewhat greater following GBR + xDBM; however, delayed. xDBM alone did not adequately resolve the bony defect. In conclusion, GBR results in rapid, clinically relevant bone closure of dental implant dehiscence defects. Adjunctive implantation of xDBM does not appear to significantly improve the healing response in the model used. PMID- 11429944 TI - Sinusfloor elevation and grafting with autogenous iliac crest bone. AB - Insufficient bone height in the posterior area of the maxilla, due to expansion of the maxillary sinus and atrophic reduction of the alveolar process of the maxilla, represents a contra-indication for insertion of dental implants. This anatomic problem can, in many cases, be solved by augmentation of the floor of the maxillary sinus. This surgical technique was introduced by Tatum. The so called top hinge door method creates a new floor of the maxillary sinus at a more cranial level. Underneath this new floor the existing space is filled with a bone graft. Implantation in the alveolar process with increased bone height allows insertion of dental implants. This sinus grafting technique was used in the present study. In total, 62 sinusfloor elevations were performed with cancellous iliac bone grafts in 42 patients. In those 62 augmented sinuses, 161 ITI screw type implants were inserted. The follow-up was 1-6 years after implantation. In 2 cases infections occurred. One implant needed an extended integration time. No implants were lost. The ITI solid screw implant appears to be a suitable implant following sinusfloor elevation operations, due to its rough surface, its shape and the size of the thread. The sinusfloor elevation procedure with autogenous cancellous bone graft appears to be a valuable and reliable pre-implantological procedure, provided a proper pre-operative investigation and careful surgery are performed. This procedure allows dental implant placement with a high success rate. PMID- 11429945 TI - Severe acute pancreatitis: contrast-enhanced CT and MRI features. PMID- 11429946 TI - Peripancreatic arteries in thin-section multislice helical CT. AB - BACKGROUND: The normal and pathologic anatomies of the peripancreatic arteries were demonstrated using thin-section multislice computed tomography (CT). METHODS: Triple-phase dynamic CT was performed in 304 consecutive patients without pancreatic pathology and in 22 patients with pancreatic carcinoma. CT angiographies images (2.5-mm collimation, multihelical pitch 3, HQ mode) and a 1.25-mm reconstruction interval were obtained with a GE Lightspeed Qx/i at 25, 40, and 80 s after bolus injection of 130-150 mL of iodinated contrast medium at a rate of 3-5 mL/s. The images were assessed mainly by the manual cine paging method. RESULTS: The anterior and posterior arcades and the dorsal pancreatic artery were frequently visualized. However, small-caliber arteries smaller than 1.5 mm in diameter, such as the pancreaticomagna, caudal pancreatic, and transverse pancreatic arteries, were infrequently visualized. In some cases, the peripancreatic arteries were very useful for differentiating the origin of tumors. CONCLUSION: Multislice helical CT enables the recognition of small peripancreatic arteries, and evaluation of these arteries should be considered when assessing tumors in the pancreaticoduodenal region. PMID- 11429947 TI - Magnetic resonance pancreatography. PMID- 11429948 TI - Evaluation of pancreatic carcinoma with FDG PET. AB - BACKGROUND: To assess the diagnostic usefulness and clinical impact of positron emission tomography with [F-18]fluorodeoxyglucose (FDG PET) on the management of patients with known or suspected pancreatic carcinoma. METHODS: Attenuation corrected FDG PET was performed in 20 patients (12 male, eight female) with pancreatic carcinoma at the time of initial diagnosis (n = 7), for tumor surveillance after Whipple surgery (n = 11), and for reevaluation after chemoradiation therapy (n = 2). Visual analysis of PET images were correlated with the results of abdominal computed tomography (CT) and carbohydrate antigen (CA) 19-9 serum tumor marker level that were obtained within 1 month of the PET study. Diagnostic validation was by histology in nine patients and by clinical or radiologic follow-up (5-48 months) in 11 patients. Changes in therapeutic management that were prompted by PET were tabulated. RESULTS: PET was concordant with the findings of abdominal CT in 14 patients (13 true positive, 1 true negative). PET detected clinically unsuspected lung lesions, confirmed subsequently by a chest CT, in one of these 14 patients. PET was discordant with CT in six patients. PET detected tumor recurrence in three patients in this group (15% of total) with nondiagnostic CT findings and elevated CA 19-9 serology. In two of these three patients, chemotherapy with gemcitabine was initiated based on PET localization of disease. Tumor was confirmed in the remaining one of the three patients at autopsy shortly after the PET study. FDG localization in a displaced loop of bowel resulted in an apparent false-positive hepatic lesion in one of six patients in the discordant group. PET underestimated the extent of metastatic disease in the remaining two of six patients due to hyperglycemia. CONCLUSION: In patients with suspected pancreatic carcinoma at the time of initial presentation, PET is complementary to abdominal CT and allows detection of unsuspected distant metastases. In patients with suspected recurrent pancreatic carcinoma, based on elevated or rising CA 19-9 serology, PET can localize the disease when abdominal CT is nondiagnostic as a result of posttherapy anatomic alteration. Imaging evaluation with PET may impact the clinical management of patients with pancreatic carcinoma. PMID- 11429949 TI - Santorinicele in pancreas divisum: diagnosis with secretin-stimulated magnetic resonance pancreatography. AB - A Santorinicele, or cystic dilatation of the dorsal pancreatic duct at the minor papilla, is seen in a small number of patients with pancreas divisum and may indicate obstruction at the minor papilla, a risk factor for pancreatitis. We present a case of a Santorinicele that was diagnosed with secretin-stimulated magnetic resonance pancreatography and treated with minor papillotomy. PMID- 11429950 TI - Hepatic adenomatosis: gadolinium-enhanced dynamic MR findings. AB - Hepatic adenomatosis is a rarely encountered liver disease. It is an entity distinct from hepatocellular adenoma and is characterized by the presence of more than 10 hepatic adenomas. We present the multiplanar and multiphasic dynamic magnetic resonance findings of hepatic adenomatosis in a 15-year-old adolescent who developed transfusional hemosiderosis and had received anabolic steroids for the treatment of aplastic anemia. PMID- 11429951 TI - Hepatic MR imaging: comparison of 2D and 3D gradient echo techniques. AB - BACKGROUND: To compare fat-suppressed three-dimensional gradient echo (3D-GRE) and two-dimensional gradient echo (2D-GRE) techniques for hepatic magnetic resonance imaging. METHODS: For 107 patients with suspected focal hepatic lesions, unenhanced 2D-GRE multislice in-phase and opposed-phase images and 3D GRE images were obtained. Two radiologists independently and in random order reviewed the images. Quantitative analysis compared contrast-to-noise ratio (C/N) of liver masses and of the liver to spleen and signal-to-noise ratio (S/N) of the liver. Subjective grading of lesion conspicuity, confidence for lesion detection, artifacts, and overall image quality were also evaluated. RESULTS: Conspicuity and confidence for focal lesions was statistically significantly higher on 3D-GRE than on 2D-GRE images. Liver-to-spleen C/N was higher on 3D-GRE images than on in phase and opposed-phase 2D-GRE images (p < 0.01). Liver S/N was higher on in phase 2D-GRE images than on 3D-GRE and opposed-phase 2D-GRE images (p < 0.001). Lesion-to-liver C/N was higher on 3D-GRE images than on 2D-GRE images for nonsolid lesions (p < 0.05), but there was no significant difference for solid lesions. Motion and pulsation artifacts were significantly fewer on 3D-GRE images, but graininess and shading were significantly higher. Overall image quality was not significantly different for the three sequences. CONCLUSION: Fat suppressed 3D-GRE sequences can afford comparable or better image quality than 2D GRE images and may have the potential to detect more focal hepatic lesions. PMID- 11429952 TI - High-flow arterioportal fistula: treatment with detachable balloon occlusion. AB - Transarterial embolization is one of the treatment choices for symptomatic hepatic arterioportal fistula that has low mortality and morbidity. Proper selection of the technique and embolic material is very important for the success of the procedure. We present a case with high-flow arterioportal fistula treated with transarterial embolization using detachable balloons. PMID- 11429953 TI - Three-dimensional spiral CT cholangiography with minimum intensity projection in patients with suspected obstructive biliary disease: comparison with percutaneous transhepatic cholangiography. AB - BACKGROUND: To evaluate the diagnostic potential of spiral computed tomographic (CT) cholangiography with minimum intensity projection (minIP) in the diagnosis of patients with suspected biliary obstruction. METHODS: Nine consecutive patients with obstructive biliary disease were enrolled in this study. Spiral CT data (3-mm slice thickness, pitch 1-2:1) obtained 65 s after the start of contrast medium injection (150 mL Ultravist 370, 3 mL/s) were reconstructed at 1 mm intervals. Three-dimensional (3D) CT cholangiography with minIP (3D CTC) was generated with a Siemens software package. The quality of 3D CTC in its ability to demonstrate the anatomic detail, the level of obstruction, and the presence or absence of isolated hepatic segments was evaluated using percutaneous transhepatic cholangiography as a gold standard. RESULTS: In all patients, 3D CTC demonstrated dilated intrahepatic ducts up to tertiary branches. 3D CTC correctly diagnosed the level of biliary obstruction and demonstrated isolated segments in all patients. In determining the cause of biliary obstruction, one patient with hilar cholangiocarcinoma was misdiagnosed as having biliary invasion by hepatocellular carcinoma. CONCLUSION: 3D CTC with minIP can determine the level and cause of biliary obstruction. 3D CTC can be obtained from regular thin section helical CT data and may be a strong competitor against diagnostic magnetic resonance cholangiography because of its superior resolution and information on adjacent soft tissues and the duct itself. PMID- 11429954 TI - Gallbladder duplication: MR cholangiography demonstration. AB - Gallbladder duplication is a rare anatomic malformation. We present a case of gallbladder duplication in a patient who underwent laparoscopic cholecystectomy in which breath-hold magnetic resonance cholangiography showed the biliary anomaly, allowing a correct preoperative differentiation of the specific type of duplication. PMID- 11429955 TI - Spontaneous rupture of the spleen: report of two cases. AB - Spontaneous splenic rupture is rare and in most cases occurs in a spleen affected by hematologic, neoplastic, or infectious disease or as a complication of pancreatitis. We report the imaging findings in two cases of spontaneous splenic rupture due to non-Hodgkin's lymphoma and pancreatic pseudocyst. PMID- 11429956 TI - Macronodular deformity of the spleen. AB - BACKGROUND AND METHODS: Macronodular splenic deformity without a focal lesion has rarely been reported. We present three such cases and discuss the possible pathomechanism. RESULTS: There was one case of macronodular deformity of unknown cause. In this case, the liver was elongated anteriorly over the dome of the spleen, and the markedly deformed spleen mimicked a round lesion in the left lobe of the liver. Color Doppler ultrasonography showed the major intra- and perisplenic vessels to be patent. There were two cases of macronodular deformity associated with extended portal thrombosis. In both cases, portal thrombosis extended throughout the intrahepatic and extrahepatic portal systems, and this portal flow disturbance was presumed to be the cause of the splenic deformity. CONCLUSION: Although very rare, thrombosis should be sought throughout the portal system when ultrasonography shows a markedly deformed spleen. Marked splenic deformity, especially in cases with an elongated liver, may mimic a liver tumor. A good understanding of its sonographic appearance may help sonographers prevent a hazardous misdiagnosis. PMID- 11429957 TI - Splenic involvement in endemic (murine) typhus: CT findings. AB - Murine typhus is a rickettsial infection that remains endemic in parts of Texas and California. Clinical manifestations of the infection are due to vasculitis. In a patient who acquired the infection in the United States, computed tomography demonstrated intrasplenic pseudoaneurysms, infarcts, and hemorrhage. PMID- 11429958 TI - Liposarcoma of the diaphragm: CT and sonographic appearances. AB - Malignant tumors arising from the diaphragm are exceedingly rare. We describe the first case, to our knowledge, of a primary diaphragmatic liposarcoma and demonstrate computed tomographic and sonographic findings. PMID- 11429959 TI - Glomus tumor of the stomach: CT findings. AB - We present the radiologic findings of gastric glomus tumors in two patients, in whom upper gastrointestinal series and computed tomography (CT) were primarily used for diagnosis. The diagnosis was surgically confirmed. Contrast-enhanced CT showed peripheral nodular or homogeneous strong enhancement in the arterial phase and prolonged enhancement in the delayed phase. PMID- 11429960 TI - Gastric volvulus associated with congenital diaphragmatic hernia, wandering spleen, and intrathoracic left kidney: CT findings. AB - We present an unusual case of gastric volvulus associated with wandering spleen, a delayed manifestation of congenital diaphragmatic hernia and left intrathoracic kidney. Gastric volvulus should be considered in any infant with unexplained vomiting and left diaphragmatic anomaly: in these patients, developmental disorders of the peritoneal visceral attachments of the left upper abdomen may coexist. The absence of ligamentous connections between the stomach, posterior abdominal wall, and spleen result in wandering spleen. We emphasize prompt surgical therapy to avoid gastric and splenic necrosis. Radiologic findings and the appearance of this complex congenital malformation are reported. PMID- 11429961 TI - Intestinal Behcet's disease: breath-hold MR imaging. AB - BACKGROUND: To determine the feasibility of magnetic resonance (MR) imaging in the evaluation of intestinal Behcet's disease. METHODS: Eight patients diagnosed to have intestinal Behcet's disease prospectively underwent MR imaging. Five patients had previously undergone abdominal surgery for intestinal Behcet's disease 27-81 months previously. Breath-hold T2-weighted single-shot fast spin echo, T1-weighted multiplanar spoiled gradient recalled echo, and postgadolinium enhanced fat-suppressed T1-weighted opposed-phase multiplanar spoiled gradient recalled echo images were obtained. MR findings were analyzed by three independent radiologists in terms of the presence and location of bowel wall thickening and ulcer, maximal bowel wall thickness, grade of bowel wall enhancement, and presence and nature of extraluminal manifestations. K statistics were used to evaluate interobserver variability. RESULTS: Seven cases (88%), including the five cases with previous operations, demonstrated bowel thickening of more than 9 mm and increased enhancement. Ulcer was depicted in four cases (50%) in the terminal ileum or an anastomosis site. Three cases (38%) showed extraluminal manifestation, such as mesenteric infiltration around the involved bowel (two cases) and a sinus tract to subhepatic abscess from perforation (one case). CONCLUSION: Breath-hold MR imaging is useful for the evaluation of bowel wall thickening, enhancement, mesentery infiltration, and abscess formation. Characteristic ulcerative lesions may be also depicted on T1- and T2-weighted breath-hold images. MR imaging is a useful method for postoperative follow-up study for the evaluation of recurrence. PMID- 11429962 TI - Abdominal cystic tumors containing small amount of fat in the septa: report of two cases. AB - We present two cases of abdominal cystic tumors containing small amounts of fat in their septa. Although the final pathologic diagnoses of these tumors were cystic lymphangioma and angiomatosis, the computed tomographic and magnetic resonance imaging features were almost identical and indistinguishable; a purely cystic mass around the region of the pancreas head associated with little mass effect on the surrounding organs and septa containing a radiologically evident fatty component. Radiologists should be aware of these two entities as differential diagnoses of abdominal cystic masses containing small amounts of septal fat. PMID- 11429963 TI - Tuberculous peritonitis: a diagnostic challenge. AB - Tuberculous (TB) peritonitis rarely occurs in developed countries. The clinical presentation and laboratory tests are usually insufficient for diagnosing TB peritonitis and distinguishing it from peritoneal carcinomatosis. We present two cases of TB peritonitis and illustrate the computed tomographic findings of the disease according to the world literature. Recognition of the radiologic manifestations and maintenance of a high index of suspicion in population at risk have paramount importance for diagnosing this uncommon disease. PMID- 11429964 TI - Psoas abscess secondary to tuberculous lymphadenopathy: case report. AB - Tuberculous psoas abscesses, a well-known sequel of tuberculous spondylitis, very rarely develop without concomitant spinal involvement. We report a unique case where a psoas abscess was secondary to retroperitoneal tuberculous lymphadenopathy in a 13-year-old boy who had no demonstrable findings of spinal tuberculosis. Computed tomography showed an obvious communication between the necrotic and calcified retroperitoneal lymph node and the psoas abscess. To the best of our knowledge, fistulization of tuberculous lymph nodes into the psoas sheath has not been reported in the English-language literature. PMID- 11429965 TI - Virtual cystoscopy of the bladder based on CT and MRI data. AB - Carcinoma of the bladder can be a serious and complex condition that is not always easy to diagnose with radiologic methods. Numerous investigations are available to establish the diagnosis and demonstrate extravesical manifestations. This article updates the role of virtual cystoscopy based on computed tomographic (CT) data at different dose levels and magnetic resonance imaging (MRI) data in the diagnosis of the carcinoma of the bladder. The appearances of normal bladder and polypoid bladder carcinoma are shown. Factors for improvement of both virtual procedures are presented and pitfalls are discussed. New developments in CT, MRI, and postprocessing procedures are discussed. The value of virtual cystoscopy in sizing, localizing, and staging of bladder carcinoma is described and compared with cystoscopy, axial CT, and MRI. PMID- 11429966 TI - Isolated bladder metastases from stomach cancer: CT demonstration. AB - Isolated metastatic tumors of the urinary bladder are extremely rare. We report three cases of isolated bladder metastasis from stomach cancer, which were proved at transurethral biopsy or total cystectomy. The lesions appeared as focal or diffuse thickening of the bladder wall on computed tomography. No other metastatic lesion was detected on imaging studies, which made the diagnosis of bladder metastasis difficult. The possibility of metastatic bladder tumor should be considered when the bladder wall is thickened focally or diffusely in a patient treated with stomach cancer. PMID- 11429967 TI - Hemangiosarcoma of the adrenal glands: CT findings in two cases. AB - Hemangiosarcomas of the adrenal gland, both benign and malignant, are exceedingly rare; only a dozen of them have been reported in literature to our knowledge. We describe herein the findings at CT of 2 primary hemangiosarcomas of the adrenal glands. We also discuss their etiology, incidence, and differential diagnosis, particularly in relation to more frequent primary adrenal lesions, such as adrenal adenomas and carcinomas. CT will generally show a heterogeneous mass with frequent necrosis, contrast-enhancement, and occasional calcifications. A prospective diagnosis cannot be made by imaging alone, and pathology represents the discriminating method for a correct diagnosis. PMID- 11429968 TI - Acute phase response in toxicity studies. II. Findings in beagle dogs injected with endotoxin or subjected to surgical operation. AB - Occurrence of characteristic transient changes in WBC counts and fibrinogen values in beagle dogs subjected to single-dose toxicity studies was pointed out in the previous survey (Hoshiya et al., 2001). These changes were thought to belong to the category of "Acute Phase Response (APR)". The purpose of the present study is to compare the APR found in the single-dose toxicity studies surveyed in our previous report with those experimentally produced by intravenous injection of 1 microgram/kg endotoxin (Experiment 1), and surgical treatment (Experiment 2) (intravenous indwelling catheterization). The animals used in Experiment 2 were intravenously injected with 1 microgram/kg endotoxin 2 weeks after the operation (Experiment 3), and the results were compared with those of Experiments 1 and 2. Each experimental group consisted of 5 dogs, and clinical, hematological and blood chemical examinations were performed. Essentially the same changes were observed in response to the intravenous injection with endotoxin and the surgical operation for intravenous indwelling catheterization in beagle dogs. The most remarkable changes common to both treatments were transient increases in the fibrinogen values and WBC counts during the 2 days from Day 1 to Day 2 of the treatment. These changes were preceded by decreases in WBC counts and fibrinogen in Experiments 1 and 3. Increased erythrocyte sedimentation rates were recorded in parallel with the increase in fibrinogen. The results obtained in the present study were similar to those found in dogs treated with various xenobiotic substances in our laboratory. These changes due to different causes were thought to belong to the category of "APR" with the same biological significance as a non-specific defense mechanism. PMID- 11429969 TI - Twenty-six-week carcinogenicity study of sulfamethoxazole in CB6F1-Tg-rasH2 mice. AB - Sulfamethoxazole (SMX), a hormone-mediated rodent-specific nongenotoxic carcinogen, was administered to CB6F1 mice carrying a human prototype c-Ha-ras gene (Tg-rasH2) at doses of 0, 25, 100 or 400 mg/kg/day and to the wild-type mice at a dose of 400 mg/kg/day in feed for 26 weeks to evaluate the carcinogenicity and to validate the Tg-rasH2 model. N-Methyl-N-nitrosourea was administered at an intraperitoneal dose of 75 mg/kg to Tg-rasH2 as a positive control and the experimental system was confirmed to be valid. Histopathological examination revealed adenomas of the lung and Harderian gland and hemangiosarcoma of the spleen at low frequencies in the Tg-rasH2 treated with SMX; however, no statistically significant differences were observed either in the onset or prevalence rates of these neoplasms compared with that in the control group. Between the wild-type mice and Tg-rasH2, the onset rate and prevalence of the neoplasms were not significantly different, but the neoplasms tended to be more frequent in Tg-rasH2 mice showing a sensitivity to tumorigenicity. Follicular epithelial cell hyperplasia was observed in the thyroid gland in the groups of Tg rasH2 given 100 mg/kg SMX or more, but no neoplastic lesion was observed. SMX was judged to be negative for carcinogenic potential in Tg-rasH2 in the present study. PMID- 11429970 TI - Application of computer-assisted sperm analysis system to elucidate lack of effects of cyclophosphamide on rat epididymal sperm motion. AB - A computer-assisted sperm analysis (CASA) system was used to examine the motion of epididymal spermatozoa derived from cyclophosphamide (CP)-treated male rats. Male rats were orally dosed daily for 1 week with 20 mg/kg of CP. Males were euthanized or were mated 3 times with untreated females at 1 day, 3 weeks, and 8 weeks after the final treatment. Significant decreases in testicular and epididymal weights and epididymal sperm counts of the treated animals were noted after 8-week recovery. Histopathological morphometry of the testis revealed minimal damage to spermatogonia at 1 day after the final treatment and to spermatocytes after 3-week recovery in the CP-treated group. On Caesarian section, increased post-implantation losses were found in females mated with CP treated males in matings starting 1 day and 3 weeks after the final treatment. On the other hand, none of the sperm motion parameters of treated males derived from the CASA system exhibited significant changes at any time points, although the spermatozoa of treated males at 1 day and 3 weeks after the final treatment were damaged at the DNA level, and the spermatozoa of males after 8-week recovery had been the target cells of CP when they were spermatogonia in the testis. It was thus found that damaged spermatozoa could exhibit no changes on their motion when the damage was confined to the nuclei, and that the effect of CP on sperm nuclei was reversible. PMID- 11429971 TI - Dose-threshold for thyroid tumor-promoting effects of orally administered kojic acid in rats after initiation with N-bis(2-hydroxypropyl) nitrosamine. AB - In order to evaluate the threshold dose of thyroid tumor-promoting effects of KA, male F344 rats were initiated with N-bis(2-hydroxypropyl) nitrosamine (DHPN; 2000 mg/kg body wt., single s.c. injection) and, starting 1 week later, received pulverized basal diet containing 0%, 0.002%, 0.008%, 0.03%, 0.125%, 0.5% or 2%KA for 20 weeks. Five rats each in the 0%, 0.125%, 0.5% and 2%KA groups were sacrificed at week 12, and 10 rats each in all groups at week 20. As an additional experiment, three groups without DHPN initiation received basal diet, a diet containing 0.5% or 2%KA for 20 weeks. The serum T4 levels were significantly decreased in the DHPN-initiated groups given 0.125%KA or more at week 12. No significant decreases in serum T3 levels were observed in the groups treated with DHPN + KA and a significant increase was evident in the 2%KA-alone group at week 20. Some rats in the DHPN + 2%KA group at weeks 12 and 20 and the 2%KA-alone group at week 20 showed pronounced elevation of serum TSH. Thyroid weights were significantly increased in the DHPN-initiated groups receiving 0.5% and 2%KA at weeks 12 and 20 and in the 2%KA-alone group at week 20. Histopathologically, the incidences of focal thyroid follicular cell hyperplasias in the DHPN-initiated groups treated with 0.125%, 0.5% and 2%KA at week 20 were 5/10, 10/10 and 8/8 rats, respectively. At week 20, adenomas were observed in 7/10 rats in the DHPN + 0.5%KA group and 8/8 rats in the DHPN + 2%KA group, and carcinomas were observed in 6/8 rats in the DHPN + 2%KA group. In the groups without DHPN initiation, only focal follicular cell hyperplasia was observed in 1/9 rats in the 2%KA-alone group. These results suggest that the no-observed adverse effect for the thyroid tumor-promoting effect of KA is 0.03% (15.5 mg/kg/day) under the present experimental conditions, and that KA possesses weak tumorigenic activity in rats due to continuous serum TSH stimulation by a non genotoxic mechanism. PMID- 11429972 TI - Acute phase response in toxicity studies. I. Survey of beagle dogs subjected to single-dose toxicity studies. AB - In the field of routine single-dose toxicity studies, we occasionally meet with transient leukocytosis associated with an increase in fibrinogen in beagle dogs within a few days after treatment with the test article. Only a little is known, however, about the toxicological significance of these changes. However, these changes were thought to belong to the category of "Acute Phase Response, APR," which has been known for a long time in connection with injury, trauma or infection. Aiming at proper understanding of these experiences, we surveyed 25 single-dose toxicity studies (7 intravenous bolus, 5 intravenous infusion, 12 oral and 1 subcutaneous treatment, hereafter referred to simply as i.v. bolus, i.v. infusion, oral and s.c.) in beagle dogs, provided with data from hematological examinations. We set the following criteria as a positive response in the present survey: increases of 50% or more in either or both WBC or fibrinogen compared to the predosing value, transiently from Day 1 to Day 3 of the study. Among 25 studies surveyed, about 1/2 of the studies exhibited increases of 50% or more in either or both fibrinogen or WBC counts compared to the predosing values showing dose-dependency transiently on Day 1 or Day 2. These changes were remarkable after intravenous application. Oral application produced similar effects, although the incidence and severity were low compared to the i.v. routes. Regarding blood chemical and hematological changes other than changes in fibrinogen and WBC counts, there were no essential differences between the groups of studies with and without the changes in fibrinogen and WBC counts. These changes were thought to be characteristic and to have occurred as incidents unrelated to other changes. The reported changes seen in single-dose toxicity studies may belong to the category of APR as the non-specific mechanism of living bodies as stated by Burns et al. (1996). PMID- 11429973 TI - Modification of eating attitudes and behavior in adolescent girls: A controlled study. AB - OBJECTIVE: The aim of this study was to evaluate the effectiveness of a school based eating disorder prevention program designed to reduce dietary restraint and concern about shape and weight among adolescent girls. METHOD: A total of 474 girls aged 13-14 years received the program as part of their normal school curriculum. An assessment-only control group included 386 pupils. Measures of eating disorder features, self-esteem, and knowledge were administered before and after the intervention and at 6-month follow-up. RESULTS: Immediately following the intervention, there was a small reduction in dietary restraint and attitudes to shape and weight in the index group, whereas there was no change in the control group. This reduction was not maintained at 6-month follow-up although the dietary restraint scores of the index group remained lower than those of the control group. DISCUSSION: This prevention program achieved change in eating attitudes and behavior, although the change was modest in size and not sustained. Focusing on a high-risk subgroup of dieters might be a more fruitful primary prevention strategy. PMID- 11429974 TI - Testing the efficacy of an eating disorder prevention program. AB - OBJECTIVE: The efficacy of an eating disorder prevention program was tested in a group of preadolescent females. Improvements in self-esteem and eating behavior were predicted relative to a control group. METHOD: Two interventions lasting 5 weeks were designed and then administered to two groups of preadolescent females. Self-esteem, body esteem, dietary restraint, and eating pathology were measured before, immediately, and 6 months following an eating disorders prevention program (n = 16) and a similarly designed control program (n = 13) focusing on fruit and vegetable intake. RESULTS: Dietary restraint in both groups decreased following the interventions. Self-esteem improved in the experimental group, but this was only marginally significant. DISCUSSION: Eating disorder prevention programs may be most effective when targeted at older, more at-risk girls. However, encouraging healthy eating in both groups of preadolescent girls reduced dietary restraint, which may be an important step in the prevention of later eating disorders. PMID- 11429975 TI - E-mail as a therapeutic adjunct in the outpatient treatment of anorexia nervosa: Illustrative case material and discussion of the issues. AB - OBJECTIVE: To explore using adjunctive e-mail in treating anorexia nervosa. METHOD: Four treatment plans were supplemented by obligatory e-mail emphasizing patients' reports of specific eating-related behaviors. Office visits occurred weekly to once every few months. E-mail contact occurred once to several times per week. Patients also saw other providers. RESULTS: All patients have shown good clinical improvement. Patients provided their assessments of contributions made by the use of e-mail and generally found it to be helpful. DISCUSSION: E mail has had excellent patient acceptability and adherence. Benefits have been attributed to increased therapeutic contact, "talking" on demand, and having to frequently confront one's integrity, honesty, and eating behaviors. Little clinician time is needed for reading and responding to e-mails. Informed consent and confidentiality issues must be carefully addressed. Controlled trials are warranted to further evaluate the roles that adjunctive e-mail may play in clinical research and practice. PMID- 11429976 TI - Do broken relationships in childhood relate to bulimic women breaking off psychotherapy in adulthood? AB - OBJECTIVE/METHOD: The case notes of 111 women presenting consecutively to an outpatient eating disorders clinic with bulimia nervosa or atypical bulimia nervosa were reviewed for pretreatment factors that predicted dropout in a retrospective study. Dropping out was conceptualized as not just a patient characteristic but as a transaction between patient and therapist. Factors believed to influence this transaction included experiences of childhood trauma, severity of eating disorder characteristics and comorbid psychiatric symptoms, demographic characteristics, waiting times for assessment and therapy, distance traveled to the clinic, previous experience of psychiatric treatment, and Eating Disorder Inventory (EDI) and Rosenberg Self-Esteem questionnaire results. RESULTS: Witnessing parental breakup, being younger, being employed outside the home, and having previous experience of psychiatric treatment predicted dropping out in logistic regression models. Experiences of childhood trauma had a dose effect relationship with dropping out. Having lower overall severity of eating disorder characteristics may also relate to dropping out. DISCUSSION: An impaired ability to trust resulting from disturbed attachments may link childhood trauma and dropping out. PMID- 11429977 TI - Anorexia nervosa and social class. AB - OBJECTIVES: (1) To examine social class status of female patients with anorexia nervosa presenting over a 33-year period; (2) to identify any differences in clinical features between the social classes. METHOD: Retrospective survey using comprehensive clinical database of patients referred to a national specialist center for the assessment and treatment of anorexia nervosa. Social class was defined using UK Registrar General's classification of father's occupation. Statistical methods included initial univariate analyses and subsequent ordinal logistic regression. RESULTS: (1) Social class distribution was consistently weighted toward social classes 1/2. (2) Possible clinical indicators examined included low body weight, binge eating, and consequent weight-regulatory behaviors such as vomiting and laxative and diuretic misuse. These clinical features and their distribution proved to be similar across the social groups. Clinically rated quality of family relationships and types of family constellations were also consistent across the social classes. (3) Dieting prodromata and onset of the disorder occurred at younger ages in social classes 1/2. (4) A modest shift in social class distribution over time was apparent, with slightly more patients presenting post-1985 likely to come from lower social classes. Claims that the social class distribution is a product of referral patterns and acceptances are disputed. CONCLUSION: We suggest that the social class bias reflects a sociocultural influence; a product of the disorder significantly often arising as an avoidant response to the conflict between social class-related family values/attitudes and adolescent turbulence within that family. PMID- 11429978 TI - Comparison of binge eating disorder and bulimia nervosa in a community sample. AB - OBJECTIVE: This study examined the relationship between binge eating disorder (BED), a newly proposed eating disorder, and bulimia nervosa (BN). METHOD: Three groups recruited from the community were compared: women with BED (n = 150), women with purging BN (n = 48), and women with nonpurging BN (n = 14). RESULTS: The three groups did not differ significantly in education, weight or shape concern, and current or lifetime prevalence of nine major mental disorders. Women with BED, compared with women with purging BN, were older, less likely to have a history of anorexia nervosa, and less likely to have been treated for an eating disorder. Obesity was more commonly associated with BED than with either subtype of BN. DISCUSSION: Our results lend some support to BED as an eating disorder distinct from purging BN. More research is needed to clarify the position of nonpurging BN relative to BED and purging BN. PMID- 11429979 TI - Sexual violence and weight control techniques among adolescent girls. AB - OBJECTIVE: Research has linked childhood sexual abuse and eating disturbances among adult females. Less is known about whether sexual abuse in the form of dating violence has implications for deviant weight control techniques among adolescent girls. This study assesses the association between sexual violence and weight control practices among girls. We also attempt to untangle the effects of family environment from this association and to determine if weak impulse control mediates the association. METHOD: Survey data were gathered from 2,629 girls in Grades 9-12 to assess health risks. Girls responded to questions regarding dating violence, unwanted sexual contact, purging, and diet pill consumption. Logistic regression was used to assess the unique contribution of sexual violence on weight control techniques. RESULTS: Dating violence and unwanted sexual contact elevated the probability that girls would report practicing weight control techniques by 6-13%. Controls for family environment did not eliminate these associations. Weak impulse control did not significantly mediate these associations. DISCUSSION: These findings suggest that sexual violence has immediate implications as a risk factor for weight control techniques in adolescence. PMID- 11429980 TI - Perfectionism in women with binge eating disorder. AB - OBJECTIVE: This study examined self-oriented (SOP), socially prescribed (SPP), and other-oriented (OOP) perfectionism in 127 obese women with binge eating disorder (BED). METHOD: Relationships between eating disorder and general psychopathology variables and SOP, SPP, and OOP were assessed. Levels of SOP, SPP, and OOP in the BED sample were compared with those of 32 normal weight women with bulimia nervosa (BN) and 60 obese non-eating-disordered individuals (NED). Structural equation modeling (SEM) was used to test models of the maintenance of BED. RESULTS: Only SPP was significantly associated with eating disorder variables related to BED. All three groups demonstrated similar levels of SPP and OOP. BN and BED groups scored significantly higher than the NED group on SOP only. SEM resulted in two models with good fits. DISCUSSION: Further research is needed on the roles of SPP and SOP in BED and on weight and shape overconcern in BED maintenance models. PMID- 11429981 TI - Association between puberty and disordered eating, body image, and other psychological variables. AB - OBJECTIVE: To examine associations between puberty and disordered eating, body image, and other psychological variables. METHOD: Females were grouped into three categories of age at puberty onset (Study 1; N = 267) as well as three categories of self-perception of prepubertal weight (Study 2; N = 222). Participants in both studies were matched on current body mass index (BMI) across categories. RESULTS: Females who perceived themselves to be overweight prior to puberty scored significantly higher on measures of disordered eating, body dissatisfaction, asceticism, drive for thinness, impulse regulation, interoceptive awareness, and perfectionism. They also perceived their current body figure to be larger when compared with those who believed they were average or underweight prior to puberty. Age at puberty onset was not significantly associated with disordered eating, body dissatisfaction, depression, or self-esteem. However, females who entered puberty at an earlier age scored higher on measures of asceticism, drive for thinness, impulse regulation, and social insecurity. They also chose a more slender ideal body figure than those who entered at an older age. DISCUSSION: Females who believe they are overweight prior to puberty may be at risk for the development of disordered eating, body image dissatisfaction, and related problems. Age at puberty onset is not a consistent risk factor of pathological eating. PMID- 11429982 TI - Gender differences in the relation between food cravings and mood in an adult community: Results from the fleurbaix laventie ville sante study. AB - OBJECTIVE: The aim of this survey was to study food cravings and its indulgence in community adults (538 women and 506 men) and to compare nutritional parameters, weight preoccupations, and weight history between cravers and noncravers. METHOD: Cravers experienced a strong urge to eat specific foods more than once a week during the past 6 months. Food intake was estimated by a 3-day food record. RESULTS: 28% of women and 13% of men were food cravers. Cravers, especially women, were more frequently concerned about their weight than noncravers. Energy intake during snacks was higher in cravers. Less than 40% of cravers reported being hungry when they experienced cravings. Women cravers indulged their cravings as often as men. They reported more frequently negative feelings, whereas men reported more frequently positive feelings. DISCUSSION: This study shows that food craving episodes are strongly associated with mood but in a different way in women and men. PMID- 11429983 TI - Cross-Cultural Research on Anorexia Nervosa: Assumptions Regarding the Role of Body Weight. AB - OBJECTIVE: To critically examine two assumptions guiding cross-cultural research on the weight concerns of anorexia nervosa: (1) that weight concerns are specific to contemporary, Western manifestations of the disorder and (2) that the dissemination of Western values regarding thinness is primarily responsible for the development of anorexia nervosa in non-Western contexts. METHOD: A review of theoretical and empirical literature on cross-cultural aspects of anorexia nervosa and the medical records of 14 Asian patients treated for eating disorders in Sydney, Australia. RESULTS AND DISCUSSION: Regarding the first assumption: It is argued that weight concerns when defined as weight loss that is positively valued (rather than a fat phobia) is a defining characteristic of anorexia nervosa and is not limited to contemporary, Western cases of the disorder. Regarding the second assumption: It is argued that the occurrence of anorexia nervosa in non-Western contexts cannot be solely attributed to the acceptance of Western thinness ideals because values and practices intrinsic to non-Western cultures are also likely to be etiologically relevant. PMID- 11429984 TI - The use of the sociocultural attitudes towards appearance questionnaire with middle school boys and girls. AB - OBJECTIVE: To assess the psychometric validity of versions of the Sociocultural Attitudes Towards Appearance Questionnaire (SATAQ) designed for use with middle school girls and boys. METHOD: As part of a larger study, the SATAQ was administered to 505 middle school children. To assess concurrent validity, scales concerning the use of Weight Control and Muscle Building Techniques as well as the Body Esteem Scale (BES) were also administered. Body mass index (BMI) was also calculated. RESULTS: Both the girls and boys versions of the SATAQ showed adequate validity. Two SATAQ subscales, Internalization and Awareness of societal ideals of thinness, were identified for girls. For boys, three SATAQ subscales were identified: Internalization and Awareness of a muscular, non-fat ideal and a scale tapping the the importance of a Muscular Look. The subscales and total score showed acceptable concurrent validity. Even after controlling for BMI, they contributed significant explained variance for the use of Weight Control Techniques in both boys and girls and use of Muscle Building Techniques in boys. Patterns of relationships among variables differed by gender. DISCUSSION: The results suggest that sociocultural influences play a role in the use of Weight Control and Muscle Building Techniques for middle school boys although their influence is not as strong as it is for the use of Weight Control Techniques by middle school girls. The SATAQ seems to be an adequate measure of the nature of some of these influences. PMID- 11429985 TI - Rationales for Food Refusal in Chinese Patients with Anorexia Nervosa. AB - OBJECTIVE: To study the rationales for food refusal among Chinese patients with typical and atypical anorexia nervosa. METHOD: Forty-eight consecutive patients with broadly defined anorexia nervosa underwent evaluation with a self-report rationale for food refusal questionnaire, the 12-item General Health Questionnaire (GHQ-12), the 21-item Beck Depression Inventory (BDI-21), the Hamilton Depression Rating Scale (HDRS), and other clinical assessments. RESULTS: Fat-phobic patients (N = 32) had a significantly higher premorbid body mass index than non-fat-phobic patients (N = 16), but they did not differ on other clinical parameters, GHQ-12, BDI-21, and HDRS scores. At clinical presentation, 3 months, and 1 year prior to presentation, fat phobia and stomach bloating were the most common rationales for food refusal among fat-phobic and non-fat-phobic patients, respectively. A total of 31% of fat-phobic patients endorsed non-fat-phobic rationales at the time of clinical presentation, whereas non-fat-phobic patients adhered to non-fat-phobic attributions more consistently. DISCUSSION: The rationales used by anorexic patients to explain noneating are more varied than implied in the 4th ed. of the Diagnostic and Statistical Manual of Mental Disorders and the ICD-10 Classification of Mental and Behavior Disorders: Clinical Descriptions and Diagnostic Guidelines. A broadened conceptualization of anorexia nervosa may enhance an understanding of patients' illness experiences and enliven research on eating disorders. PMID- 11429986 TI - Evidence of diffuse atrophy of the thyroid gland in patients with anorexia nervosa. AB - OBJECTIVE: The altered function of the hypothalamic-pituitary-thyroid axis (HPT) in anorexia nervosa (AN) patients has not been clearly elucidated so far. Low triiodothyronine (T3) syndrome and a blunted and delayed thyrotropin (TSH) response to exogenously administrated thyrotropin-releasing hormone (TRH) are common findings. However, no attention has been paid to thyroid morphology in AN patients. METHOD: We performed an ultrasonographic (US) evaluation of the thyroid gland in 22 AN patients and in 44 age and sex-matched control subjects. Lean body mass (LBM) was determined by dual-energy X-ray absorptiometry. RESULTS: US determined thyroid volume was significantly reduced in AN patients (9.2 +/- 0.4 vs. 17.8 +/- 1.2 ml in the controls; M +/- SEM; p < 10(-4)). In healthy subjects, it has previously been established that thyroid volume can be estimated from age and body weight. In the present study, the measured thyroid volume in patients with AN was significantly lower than the predicted thyroid volume (measured: 9.2 +/- 0.4 vs. estimated: 12.2 +/- 0.2 ml; p < 10(-4)). Furthermore, in the AN patients, the thyroid size was not correlated to body mass index (BMI) or LBM. DISCUSSION: Thyroid volume in AN patients was markedly reduced compared with the control group and with the volume expected from age and body weight or LBM. This indicates thyroid atrophy, which, hypothetically, could be involved in a vicious circle maintaining anorectic or depressive symptomatology. PMID- 11429987 TI - Evidence for long-term pancreatic damage caused by laxative abuse in subjects recovered from anorexia nervosa. AB - OBJECTIVE: This study examined whether a prior history of laxative abuse results in long-term changes in gastrointestinal function. METHOD: The functioning of the enteroinsular axis was examined by measuring the insulin response to a standard meal. The study involved 18 subjects who had fully recovered from anorexia nervosa (AN) and an age and weight-matched control group. In the recovered group, 10 of 18 subjects had a history of laxative abuse. RESULTS: Subjects with a prior history of laxative abuse show a more gradual increase and decrease in insulin secretion, but no differences in glucose response or hunger ratings. DISCUSSION: Because there are no differences in the glucose response to the meal, it is hypothesized that the difference in insulin response is due to changes in the enteroinsular axis. These data indicate that chronic laxative abuse induces long term changes in gastrointestinal function. PMID- 11429988 TI - Effect of gender composition of school on body concerns in adolescent women. AB - OBJECTIVE: This study aimed to investigate the role of gender composition of school on body figure preferences, eating disorder symptomology, and role concerns. METHOD: Questionnaires were completed by 261 Australian adolescent girls in two private single-sex and two private coeducational school environments. RESULTS: There was no difference in nominated ideal figure or eating disorder scores between the schools. However, girls in the single-sex schools placed a greater emphasis on achievement than their counterparts at the coeducational schools. These role concerns had a differential impact on prediction of the ideal figure, whereby the importance placed on intelligence and professional success predicted the choice of a thinner ideal figure for the single-sex schools, but a larger ideal for the coeducational schools. DISCUSSION: It was concluded that the motivation for thinness differs between single-sex and coeducational schools. PMID- 11429989 TI - Georgamide, a new cyclic depsipeptide with an alkynoic acid residue from an Australian cyanobacterium. AB - A cyclic depsipeptide, georgamide (1), was isolated from an Australian cyanobacterium and characterized by spectroscopic means. The constituent units were five amino acid residues, one each of L-Thr, L-Pro, L-Val, N-Me-L-Val, and N Me-L-Phe, and two hydroxy carboxylic acids, 2(S)-hydroxy-3(R)-methylpentanoic acid and the unusual 2,2-dimethyl-3-hydroxy-7-octynoic acid. The stereochemistry was determined by hydrolysis of the peptide followed by derivatization and HPLC comparison with standard samples. PMID- 11429990 TI - Cancer chemopreventive agents. New depsidones from Garcinia plants. AB - In a search for cancer chemopreventive agents from natural sources, chemical constituents of two kinds of Garcinia plants, Garcinia neglecta and Garcinia puat, collected in New Caledonia, were examined. Five new depsidones, garcinisidone-B (2), -C (3), -D (4), -E (5), and -F (6), were isolated, and their structures were determined by spectrometric analyses. Inhibitory effects of these depsidones on EBV-EA activation induced by TPA in Raji cells were also demonstrated. PMID- 11429991 TI - Two novel cyclic peptides with antifungal activity from the cyanobacterium Tolypothrix byssoidea (EAWAG 195). AB - Two novel cyclic tridecapeptides, tolybyssidins A (1) and B (2), were isolated from the culture medium of mass cultured cyanobacterium Tolypothrix byssoidea (EAWAG 195) by means of bioguided isolation. The gross structures of these peptides were determined by 1D and 2D NMR experiments and tandem mass spectrometry. Both peptides contain the nonnatural amino acid dehydrohomoalanine (Dhha) as well as proteinogenic amino acids albeit with D- or L-configuration. The compounds exhibit moderate antifungal activity against the yeast Candida albicans. PMID- 11429992 TI - New tirucallane-type triterpenes from Dysoxylum variabile. AB - Eight new tirucallane-type triterpenes, dyvariabilins A-H (1-8), three known tirucallanes, niloticin (9), dihydroniloticin (10), and tirucalla-7,24-diene-3 beta,23-diol (11), and two known sesquiterpenes, 1-(1-hydroxy-2-methylpropyl)-3a methyl-7-methyleneoctahydroinden-4-ol and (+)-aphanamol I, were isolated from the stem bark of Dysoxylum variabile. Tirucallanes 1 and 11 with an allylic hydroxy group in the side chain and dyvariabilin C (3) with an alpha-epoxy group at positions 7 and 8 showed high instability in acidic medium and formed five hitherto unknown semisynthetic tirucallanes. Dyvariabilins B (2) and C (3) as well as the mixtures of dyvariabilins E and F (5 and 6) and dyvariabilins G and H (7 and 8) showed weak cytotoxicity against KB cells. PMID- 11429993 TI - Sequences and antimycoplasmic properties of longibrachins LGB II and LGB III, two novel 20-residue peptaibols from Trichoderma longibrachiatum. AB - Longibrachins are members of the class of natural Aib-containing peptides designated as peptaibols. Six longibrachins, LGA I-IV and LGB II and III, were purified from a Trichoderma longibrachiatum strain by a procedure employing several chromatography steps including reversed-phase HPLC. The amino acid sequence determination was based on a combination of liquid secondary ion mass spectrometry (LSIMS) and two-dimensional 1H and 13C NMR spectroscopy. Longibrachins are 20-residue peptaibols with a C-terminal phenylalaninol and either neutral (LGA; Gln18) or acidic (LGB; Glu18) character. Longibrachins LGB II and III have novel sequences. Both longibrachins LGA and LGB show significant bactericidal activity against mycoplasmas (Acholeplasma, Mycoplasma, and Spiroplasma), with minimal inhibitory concentrations in the range 1.56-12.5 microM (3-25 micrograms/mL), and also perturb the permeability of membrane bilayers. Longibrachin LGA IV is the most potent of the presently known 18-20 residue peptaibols. The antimicrobial and membrane-perturbing properties of longibrachins, which are described here for the first time, were shown to be correlated. PMID- 11429994 TI - Acetylated analogues of the microtubule-stabilizing agent discodermolide: preparation and biological activity. AB - A series of eight discodermolide acetates have been prepared using natural (+) discodermolide and evaluated for in vitro cytotoxicity against the cultured murine P-388 leukemia cells. The acetylated analogues showed a significant variation of cytotoxicity and suggested the importance of C-11 and C-17 hydroxyl groups for potency. The preparation and structure elucidation of the new analogues are described. PMID- 11429995 TI - New triterpenoids from the Red Sea sponge Siphonochalina siphonella. AB - Clear differences were found when comparing the chemical content of the northern, Gulf of Eilat, to the southern-central, Dahlak archipelago, Red Sea sponge Siphonochalina siphonella. The Dahlak sponge was found to be richer in the more polar triterpenes. Nine new compounds (1-4, 7, 8, 15-17) were isolated and identified, among them two sipholane glycosides, sipholenoside A and B (7 and 8), and one compound, dahabinone A (17), possessing a new skeleton. PMID- 11429996 TI - Cytotoxic flavonoids with isoprenoid groups from Morus mongolica. AB - A new pyranoflavanone, sanggenol L (1), a Diels-Alder type adduct regarded as a cycloaddition product of a dehydrogeranylflavanone and a prenylchalcone, sanggenol M (2), along with four new 2-arylbenzofurans with isoprenoid units, mulberrofurans W-Z (3-6), were isolated together with 10 known flavonoids from Chinese Morus mongolica. The structures of these novel compounds were elucidated by spectroscopic methods. All flavanones investigated here showed higher cytotoxicity against human oral tumor cell lines (HSC-2 and HSG) than against normal human gingival fibroblasts (HGF). Among them, the cytotoxicity of compound 2 and the Diels-Alder type flavanone sanggenon C (7) isolated from Morus cathayana were the most potent. On the other hand, seven 2-arylbenzofurans exhibited lower cytotoxicity and tumor specificity as compared with flavanones. PMID- 11429997 TI - Vertilecanins: new phenopicolinic acid analogues from Verticillium lecanii. AB - Five new phenopicolinic acid analogues (1-5) have been isolated from solid substrate fermentation cultures of Verticillium lecanii. The most abundant component (vertilecanin A; 1) displays antiinsectan activity against Helicoverpa zea. These compounds were obtained by chromatographic fractionation of the EtOAc culture extract and identified by analysis of NMR and MS data. The known fungal metabolites 2-decenedioic acid and 10-hydroxy-8-decenoic acid were also isolated from these cultures. PMID- 11429998 TI - Hydroxylation of yohimbine in superacidic media: one-step access to human metabolites 10 and 11-hydroxyyohimbine. AB - Two major human metabolites of yohimbine (1), 10- and 11-hydroxyyohimbine (2 and 3), were prepared by direct hydroxylation of 1 under superacidic conditions. In this medium, the four positions of the benzene part of yohimbine were hydroxylated and the corresponding monohydroxylated compounds (2-5) were isolated. The structures of 2-5 were elucidated by spectroscopic methods including 1D and 2D NMR techniques. PMID- 11429999 TI - A facile synthetic approach to prenylated flavanones: first total syntheses of (+/-)-bonannione A and (+/-)-sophoraflavanone A. AB - A facile and efficient approach for the syntheses of both C-8 and C-6 prenylated flavonoids has been developed that features a highly regioselective prenylation of 2,4,6-trihydroxyacetophenone and regioselective cyclization of prenylated polyhydroxy chalcones. Thus, the first efficient total syntheses of (+/-) sophoraflavanone A (1) and (+/-)-bonannione A (2), two naturally occurring geranylated flavanones with antibacterial activities, have been achieved starting from the key intermediate 3 via regioselective cyclization of geranylated tetrahydroxychalcone 4. PMID- 11430000 TI - New lipopeptides from the Caribbean cyanobacterium Lyngbya majuscula. AB - Four new metabolites have been isolated from a marine red cyanobacterium, Lyngbya majuscula, collected at Boca del Drago Beach, Bocas del Toro, Panama. The planar structures were elucidated by 1D and 2D NMR techniques. These compounds were assigned the trivial names pseudodysidenin (2), dysidenamide (3), nordysidenin (4), and dragonamide (7). PMID- 11430001 TI - Isoaurostatin, a novel topoisomerase inhibitor produced by Thermomonospora alba. AB - A novel inhibitor of topoisomerase I designated as isoaurostatin (1) was isolated from the culture filtrate of Thermomonospora alba strain No. 1520. The structure of 1 was determined to be 6,4'-dihydroxyisoaurone on the basis of spectroscopic (NMR and MS) methods. Compound 1 inhibited the relaxation activity of calf thymus topoisomerase I in a noncompetitive manner and did not inhibit the relaxation and decatenation of human placenta topoisomerase II. Compound 1 is an inhibitor belonging to cleavable complex-nonforming type without DNA intercalation. PMID- 11430002 TI - Eleven novel diarylheptanoids and two unusual diarylheptanoid derivatives from the seeds of Alpinia blepharocalyx. AB - An EtOH extract of the seeds of Alpinia blepharocalyx afforded 11 novel diarylheptanoids, named deoxycalyxin A (1), epicalyxin F (2), calyxin K (3), epicalyxin K (4), calyxin I (5), epicalyxin I (6), calyxin J (7), epicalyxin J (8), and calyxin L (9), an epimeric mixture of calyxin M (10) and epicalyxin M (11), and two unusual diarylheptanoid derivatives, named neocalyxins A (12) and B (13), together with four known calyxins, calyxins A (14), F (15), E (16), and G (17). Structures were elucidated by spectroscopic techniques including 2D NMR spectroscopy. All compounds were examined for cytotoxicity toward murine colon 26 L5 carcinoma and human HT-1080 fibrosarcoma cells. Diarylheptanoids 2, 3, and 5 were cytotoxic against both cell lines, while 4 and 6-8 were cytotoxic against human fibrosarcoma cells. PMID- 11430003 TI - Studies on flavans. 1. Facile synthesis of (+/-)-7-hydroxy-3',4' methylenedioxyflavan and (+/-)-4'-hydroxy-7-methoxyflavan by a BF3.Et2O-mediated pyran cyclization. AB - A facile approach for the synthesis of flavans was developed by employing a BF3.Et2O-catalyzed pyran cyclization in an aprotic polar solvent as a key step, by which concise total syntheses of (+/-)-7-hydroxy-3',4'-methylenedioxyflavan (1) and (+/-)-4'-hydroxy-7-methoxyflavan (2), two naturally occurring flavans, were achieved. PMID- 11430004 TI - A new diterpene from Hypoestes serpens. AB - From a defatted chloroform extract of the aerial parts of Hypoestes serpens a new diterpene exhibiting relaxant activity on isolated rat aorta was obtained. The structure of this compound, named serpendione (1), was fully established by the interpretation of its spectral data. PMID- 11430005 TI - New bioactive sulfated metabolites from the Mediterranean tunicate Sidnyum turbinatum. AB - In addition to the known sodium 3,7,11,15-tetramethylhexadeca-1,19-diyl sulfate (4), the BuOH extract of the Mediterranean tunicate Sidnyum turbinatum was shown to contain four new metabolites: 1-heptadecanyl sulfate (1), 1-octadecanyl sulfate (2), sodium (2S)-2,6,10,14-tetramethylpentadeca-1,18-diyl sulfate (3), and 1-hexyl sulfate (5). Their structures were determined by spectroscopic and chemical methods. Compounds 1-5 exhibited in vitro antiproliferative activity estimated on the WEHI 164 cell line. PMID- 11430006 TI - Microbial transformation of ent-kaurenoic acid and its 15-hydroxy derivatives by the SG138 mutant of Gibberella fujikuroi. AB - Feeding experiments with ent-kaurenoic acid (4), 15 alpha-hydroxy-ent-kaurenoic acid (5), 15 beta-hydroxy-ent-kaurenoic acid (6), and mixtures of 4 plus 5 and 4 plus 6 were conducted using the SG138 mutant of Gibberella fujikuroi, to gain information about the phenotype of this unique strain. The biotransformation of 5 gave 7 beta,15 alpha-dihydroxykaurenolide (9) and 7 beta,15 alpha-dihydroxy-ent kaurenoic acid (13). The incubation of 6 produced 7 beta,15 beta-dihydroxy-ent kaurenoic acid (7) and 7 beta,15 beta-dihydroxykaurenolide (10). No 15 hydroxylated gibberellins were detected in any of these experiments. The results indicated that a hydroxy group at C-15 does not inhibit 7 beta-hydroxylase activity but in the SG138 strain obstructs the enzymatic ring-B contraction of ent-kaurenoids to gibberellins. Exogenous 4 stimulated both the excretion of ent kaurene and the fungal metabolism of 5 and 6. PMID- 11430007 TI - Aplyzanzine A, a new dibromotyrosine derivative from a Verongida sponge. AB - Aplyzanzine A (1), a novel bisdibromotyrosine derivative, has been isolated from the Indo-Pacific sponge Aplysina sp. Its structure was elucidated mainly on the basis of 1D and 2D NMR and MS spectroscopic data. PMID- 11430008 TI - Merilactone, an unusual C19 metabolite from the root extract of Chiococca alba. AB - A C19 metabolite has been isolated from the methanol extract of the roots of Chiococca alba and characterized as a new nor-seco-pimarane, to which we have given the trivial name merilactone (1). The structure and relative stereochemistry of 1 was established by spectral data interpretation. PMID- 11430009 TI - Antisweet saponins from Gymnema sylvestre. AB - Three new oleanane-type triterpene glycosides (1-3), along with the sodium salt of alternoside II (4), were isolated from an ethanol extract of the leaves of Gymnema sylvestre. The structures of these new saponins were identified as 21 beta-O-benzoylsitakisogenin 3-O-beta-D-glucopyranosyl(1-->3)-beta-D glucuronopyranoside (1), the potassium salt of longispinogenin 3-O-beta-D glucopyranosyl(1-->3)-beta-D-glucuronopyranoside (2), and the potassium salt of 29-hydroxylongispinogenin 3-O-beta-D-glucopyranosyl(1-->3)-beta-D glucuronopyranoside (3). The aglycon of 3, gymnemagenol (3a), was characterized as 3 beta,16 beta,28, 29-tetrahydroxyolean-12-ene. Structure elucidation was accomplished by interpretation of NMR (DQF-COSY, HMQC, and HMBC) results, FABMS, and hydrolysis. Saponin 1 and the sodium salt of alternoside II (4) exhibited antisweet activity. PMID- 11430010 TI - Colossolactones, new triterpenoid metabolites from a Vietnamese mushroom Ganoderma colossum. AB - Seven new triterpenoid metabolites (colossolactones; 1-7) were isolated from a fruiting body of Ganoderma colossum, and their structures were determined by MS and NMR methods. PMID- 11430011 TI - Bidentatoside I, a new triterpene saponin from Achyranthes bidentata. AB - Bidentatoside I (1) is a new triterpene saponin bearing an unusual dioxopropionic acid unit, isolated from the roots of Achyranthes bidentata. Structural elucidation was performed mainly by chemical and homo- and heteronuclear 2D NMR techniques. This compound did not show any potentiation of the in vitro cytotoxicity of cisplatin in the HT 29 human colon cancer cell line. PMID- 11430012 TI - Cytotoxic polyacetylenes from the twigs of Ochanostachys amentacea. AB - Bioassay-guided investigation of the twigs of Ochanostachys amentacea using LNCaP (hormone-dependent human prostate cancer) cells as a monitor led to the isolation of three alkynes, the known (S)-17-hydroxy-9,11,13,15-octadecatetraynoic acid (minquartynoic acid, 1) and two novel analogues, (S)-17,18-dihydroxy-9,11,13,15 octadecatetraynoic acid (2) and (S)-17-hydroxy-15E-octadecen-9,11,13-triynoic acid (3). Compounds 1-3 were tested against a panel of human tumor cell lines and found to be significantly cytotoxic. PMID- 11430013 TI - A novel anti-HIV macrocyclic peptide from Palicourea condensata. AB - A 37 amino acid cyclic polypeptide has been isolated from the organic extract of the tropical tree Palicourea condensata. Palicourein (1) is the largest of a growing family of plant peptides that contain a cyclized amino acid backbone cross-linked via three internal disulfide bridges. Palicourein inhibits the in vitro cytopathic effects of HIV-1RF infection of CEM-SS cells with an EC50 value of 0.1 microM and an IC50 value of 1.5 microM. PMID- 11430014 TI - Isolation of symlandine from the roots of common comfrey (Symphytum officinale) using countercurrent chromatography. AB - Three pyrrolizidine alkaloids, symlandine, symphytine, and echimidine (1-3), were isolated from the roots of Symphytum officinale using a one-step countercurrent chromatography procedure. The structures of 1-3 were confirmed by several spectroscopic techniques including 2D NMR methods. This is the first description of the separation of symlandine (1) from its stereoisomer, symphytine (2). PMID- 11430015 TI - Granulatimide and 6-bromogranulatimide, minor alkaloids of the Brazilian ascidian Didemnum granulatum. AB - Reinvestigation of the extract of the ascidian Didemnum granulatum collected on the Brazilian coastline led to the isolation of two minor compounds, granulatimide (2) and 6-bromogranulatimide (3), which have been identified by analysis of their spectroscopic data. The isolation of 2 and 3 from D. granulatum corroborates previous assumptions about the occurrence of granulatimide as a natural product. PMID- 11430016 TI - Characterization of novel methyl-branched chain fatty acids from a halophilic Bacillus species. AB - The 4-methylated fatty acids 4,9-dimethyldecanoic, 4,11-dimethyldodecanoic, 4,10 dimethyldodecanoic, and 4,13-dimethyltetradecanoic acid as well as the 2 methylated fatty acids 2,13-dimethyltetradecanoic and 2,12-dimethyltetradecanoic acid were identified for the first time in nature in the halophilic bacterium Bacillus sp. isolated from the salt pans of Burgas in Bulgaria. The principal fatty acids in this bacterium were a series of iso-anteiso fatty acids with chain lengths between C11 and C19, but an interesting series of linear alkylbenzene fatty acids with chain lengths between C10 and C14, such as 12-phenyldodecanoic acid, were also identified. The novel 4-methylated fatty acids were characterized using a combination of GC-MS and chemical transformations such as N acylpyrrolidide derivatization. The 2-methylated fatty acids were also identified by GC-MS and gas chromatographic coelution with synthetic samples. The novel methyl-branched fatty acids probably originated from the selective incorporation of methylmalonyl-CoA by one of the fatty acid-synthesizing enzymes of the bacterium. PMID- 11430017 TI - Revised structure for a sesquiterpene lactone from Bombax malbaricum. AB - A sesquiterpene lactone isolated from Salmalia malbaricum (syn Bombax malbaricum) roots was previously identified as hemigossylic acid lactone-7-methyl ether (1). 2D NMR experiments have shown this is a new compound, isohemigossylic acid lactone-2-methyl ether (2). PMID- 11430018 TI - Three new peroxides from the sponge Plakinastrella species. AB - Two new five-membered-ring peroxide acids, plakinic acid F (3) and epiplakinic acid F (4), and a new peroxide-lactone, plakortolide F (5), were isolated from a sponge of the genus Plakinastrella collected from Felicite Island, Seychelles. The structures were elucidated through spectral analysis. The free acids 3 and 4 exhibit moderate antifungal activity against Candida albicans with minimum inhibitory concentrations of 25 micrograms/mL (SDB) and 3.1 micrograms/mL (RPMI) for 3, and 25 micrograms/mL (SDB) and 6.25 micrograms/mL (RPMI) for 4, respectively. Both also showed moderate in vitro inhibition of Aspergillus fumigatus with IC90's of 25 micrograms/mL. PMID- 11430019 TI - Natural product-based anti-HIV drug discovery and development facilitated by the NCI developmental therapeutics program. AB - During the decade 1987-1996, the Developmental Therapeutics Program (DTP) of the National Cancer Institute (NCI) provided infrastructure support for both intramural and extramural anti-HIV (human immunodeficiency virus) drug discovery research and development. This retrospective review describes some of the anti HIV lead discovery and development that took place under DTP auspices or which was substantially facilitated by resources made available through the DTP. Examples highlighted include leads identified through the initial screening of pure natural product derived compounds and those derived from bioassay-guided fractionation of crude natural product extracts, and these are classified according to the mechanism of action targeting the critical steps within the replication cycle of HIV. PMID- 11430020 TI - Highly diastereoselective 5-hexenyl radical cyclizations with Lewis acids and carbohydrate scaffolds. AB - [figure: see text] Carbohydrates as removable chiral scaffolds for free radical cyclizations were examined for the first time. This investigation illustrates the utility of two inexpensive carbohydrate derivatives as sources of asymmetry for 5 hexenyl radical cyclizations. Diastereomeric ratios as high as 100:1 were achieved with an ester-appended (+)-isosorbide hexose and 70:1 for a diol protected D-xylose pentose. Temperature dependence, Lewis acids, and solvents were all examined. By correlation with known compounds, the newly generated chiral centers were of the (S)-configuration. PMID- 11430021 TI - Highly efficient ligands for palladium-catalyzed asymmetric alkylation of ketone enolates. AB - [figure: see text] Ferrocene-modified chiral pocket ligands have been studied in the palladium-catalyzed asymmetric alkylation of simple ketone enolates, in which (R,R,Sp,Sp)-1 containing two pairs of matched chiralities, central chirality and planar chirality, behaved very efficiently in this reaction and up to 95% ee value was achieved. PMID- 11430022 TI - Formation of benzo[b]fluorenes and the benzo[a]fluorene core of the fluostatins by cyclization of diaryldiynones. AB - [figure: see text] Thermal cyclization of 1-[2-(trimethylsilylethynyl)phenyl]-3 arylpropinones was expected to give benzo[b]fluorenones. However, benzo[a] fluorenones were also formed as a result of a new rearrangement. These tetracycles possess the core structure of the fluostatins and isoprekinamycin. PMID- 11430023 TI - Novel use of imidazolium ylides in an efficient synthesis of 2-substituted imidazoles. AB - [figure: see text] A new reaction of imidazoles was discovered involving the formation of an imidazolium ylide, which on trapping with various electrophiles afforded diverse 2-substituted imidazoles. The facile, convenient reaction conditions when compared to the existing procedures make this reaction the method of choice in the preparation of 2-substituted imidazoles. Moreover, the reaction differs from the reported methods since the products (viz., 1) contain an alpha substituent that is transformed by solvolysis chemistry into further functionalized derivatives. PMID- 11430024 TI - Chiral P,N-ligands based on ketopinic acid in the asymmetric Heck reaction. AB - [figure: see text] Novel chiral P,N-ligands were synthesized from (1S)-(+) ketopinic acid using palladium-catalyzed coupling reaction of a vinyl triflate and either a diarylphosphine or a dialkylphosphine as the key step. Palladium complexes of these ligands are efficient catalysts for asymmetric Heck reaction between aryl or alkenyl triflates and cyclic alkenes. Products were obtained with good to excellent enantioselectivity from arylation and alkenylation of 1,2 dihydrofuran, cyclopentene, and 4,7-dihydro-1,3-dioxepin. PMID- 11430025 TI - Lanthanide trifluoromethanesulfonate-catalyzed asymmetric aldol reactions in aqueous media. AB - [figure: see text] Catalytic asymmetric aldol reactions catalyzed by lanthanide trifluoromethanesulfonates in aqueous media have been realized for the first time using a chiral crown ether. PMID- 11430026 TI - Synthesis of configurationally defined sexi- and octinaphthalene derivatives. AB - [figure: see text] Configurationally defined optically active octinaphthalenes were synthesized using the oxidative coupling of optically active quaternaphthalenes with a 2-hydroxynaphthol moiety as a key reaction. The absolute configuration was determined by comparison with products of [6 + 2] coupling. PMID- 11430027 TI - Control of the arrangement of dipolar orientation of pyrimidine inside the conjugated backbone. AB - [figure: see text] Linear molecules with different arrangement of dipolar pyrimidine moieties in the pi-conjugated backbone were synthesized by a Pd catalyzed Sonogashira coupling reaction. Significant reduction of the HOMO-LUMO energy band gap and improvement of fluorescence quantum yield were observed upon incorporating pyrimidine into the conjugated backbone. PMID- 11430028 TI - Versatile 8-oxabicyclo[3.2.1]oct-6-en-3-one: stereoselective methodology for generating C-glycosides, delta-valerolactones, and polyacetate segments. AB - [figure: see text] A new rearrangement of functionalized methoxy glycosides and a regioselective as well as stereoselective intramolecular Michael addition giving delta-valerolactones and C-glycosides are described. Applications to the synthesis of marine natural products are reported. Chemoselective deprotection of benzylated hydroxy groups is assumed to be facilitated by 6-endo-tet interaction with the 1,3-dithiane functionality. PMID- 11430029 TI - Optimization of palladium-catalyzed polyene cyclizations: suppression of competing hydride transfer from tertiary amines with Dabco and an unexpected hydride transfer from 1,4-dioxane. AB - [figure: see text] This paper demonstrates that both 1,2,2,6,6 pentamethylpiperidine (PMP) and 1,4-dioxane can act as hydride donors in palladium-catalyzed polyene cyclizations of 2 and 3. Studies using PMP-d3 and dioxane-d8 either incorporate a deuterium atom into the monosubstituted product or completely inhibit the hydride transfer so that the second ring closure occurs in high yield. Dabco is the best substitute for PMP. PMID- 11430030 TI - Sulfanyl- and selanyldifluoromethylphosphonates as a source of phosphonodifluoromethyl radicals and their addition onto alkenes. AB - [figure: see text] Two different strategies are shown to produce sulfanyl and selanyldifluoromethylphosphonates. Thus, treatment of sulfanyldichloromethylphosphonates by 3HF.NEt3 in the presence of zinc bromide produces the corresponding sulfanyldifluoromethylphosphonates. In addition, lithiation of difluoromethylphosphonates followed by quenching with phenylsulfanyl chloride, phenylselanyl chloride, or diphenyl diselenide yields the corresponding sulfanyl- and selanyldifluorophosphonates. Generation of phosphonodifluoromethyl radicals from such precursors in the presence of alkenes produces the expected adducts. PMID- 11430031 TI - Enantiomerically pure cyclopropyl hemiacetals: lipase-catalyzed synthesis of chiral, nonracemic ester homoenolate equivalents. AB - [figure: see text] Enantiomerically pure cyclopropyl hemiacetals can be obtained by lipase-catalyzed kinetic resolution of their acylated congeners. It is demonstrated that lipases from Candida antarctica and Pseudomonas cepacia show enantiodivergent behavior toward these substrates. Subsequent ring opening of these building blocks can be achieved with ZnCl2 leading to chiral, nonracemic alpha-substituted homoenolate anions. PMID- 11430032 TI - Total synthesis of (-)-stellettamide B and determination of its absolute stereochemistry. AB - [figure: see text] The first total synthesis of (-)-stellettamide B has been achieved by a sequence based on amide coupling of the chiral 1-(aminomethyl) indolizidine fragment, prepared by TiCl4-mediated asymmetric allylation of the tricyclic N-acyl-N,O-acetal, with the chiral trienoic acid fragment. This synthesis led to revision of the published relative stereochemistry of the natural product and established its absolute stereochemistry to be 1S,4S,8aR,6"R. PMID- 11430033 TI - Synthesis and growth inhibitory properties of glucosamine-derived glycerolipids. AB - [figure: see text] 2-Amino C-glycerolipid 1b was synthesized by using the Ramberg Backlund rearrangement as the key step. beta-C-Glycerolipid 1b exhibits in vitro antiproliferative effects strikingly similar to those of O-glycoside analogue 1a. PMID- 11430034 TI - Selective fowler reductions: asymmetric total syntheses of isofagomine and other 1-azasugars from methyl nicotinate. AB - [figure: see text] An efficient, high-yielding strategy has been developed for the asymmetric synthesis of 1-N-iminosugars (1-azasugars), a new class of glycosidase inhibitors with promising biomedical applications. A highly regioselective procedure for the 1,2-reduction of substituted pyridines was employed to transform methyl nicotinate into several representative 1-azasugars. PMID- 11430035 TI - 195Pt NMR determination of the enantiomeric purity and absolute configuration of trisubstituted allenes by using [PtCl3(C2H4)]-[(S,S)-(1-NpMeCH)2NH2]+ as CDA. AB - [figure: see text] The ionic CDA [PtCl3(C2H4)]-[(S,S)-(1-NpMeCH)2NH2]+ produces, on exchange of its coordinated ethylene by chiral trisubstituted allenes, diastereoisomeric mixtures originating distinct 195Pt NMR resonances for the complexed enantiomers, thus allowing the determination of the enantiomeric purity. A reproducible correlation between relative positions of platinum signals due to the complexed enantiomers and their absolute configuration has been found. PMID- 11430036 TI - A general [3 + 2 + 1] annulation strategy for the preparation of pyridine N oxides. AB - [figure: see text] Stabilized ketone, aldehyde, and ester enolates react with vinamidinium hexafluorophosphate salts and hydroxylamine hydrochloride to give access to the corresponding pyridine N-oxides. The annulation reactions proceed in good to excellent yields with vinamidinium salts with a range of beta substituents (R3 = halo, aryl, nitro, trifluoromethyl). PMID- 11430037 TI - Synthesis of the macrocyclic core of laulimalide. AB - [figure: see text] A stereoselective synthesis of 3, corresponding to the fully functionalized macrocyclic core of the novel microtubule-stabilizing agent, laulimalide, has been completed. Efficient macrolactonization was achieved by a Mitsunobu reaction, installing the sensitive (Z)-enoate, and macrocyclic stereocontrol was then exploited to introduce the methyl group and trans-epoxide. PMID- 11430038 TI - Hydroboration of unsaturated phosphonic esters: synthesis of boronophosphonates and trisubstituted vinylphosphonates. AB - [figure: see text] Hydroboration of vinyl phosphonates with pinacolborane (PBH) proceeds to give phosphonoboronates, 2. Surprisingly, such compounds have not been reported before. The reaction works well with terminal alkenylphosphonates. Internal alkenylphosphonates give complex mixtures. Hydroboration/Suzuki coupling of alkynylphosphonates, with PBH, in a one-pot procedure gives trisubstituted vinylphosphonates in good overall yields and provides a new synthesis of these compounds. PMID- 11430039 TI - Alkyne cross metathesis reactions of extended scope. AB - [figure: see text] A catalyst formed in situ from Mo[N(t-Bu)(Ar)]3 1 (Ar = 3,5 dimethylphenyl) and CH2Cl2 in toluene effects cross metathesis reactions of functionalized alkynes that are beyond reach of more traditional promotors. An application to the synthesis of prostaglandin E2 (PGE2) 19 and the acetylated PGE derivative 18b shows the compatibility of this method with sensitive substrates. PMID- 11430040 TI - Monolithiocavitands: versatile intermediates for new cavitand-based hosts. AB - [figure: see text] Cavitand bowls carrying two derivatizable groups at the rim have been efficiently prepared through a selective lithium/halogen exchange electrophile quenching procedure. This methodology lends itself to the single step preparation of the first all carbon-linked bis-bowl and a new class of inherently chiral cavitands. PMID- 11430041 TI - An expedient route to 2,3-substituted and fused benzo[a]quinolizine-4-thione framework via ring annulation with beta-oxodithioesters. AB - [figure: see text] An efficient highly convergent route to hitherto unreported 2,3-substituted and annulated benzo[a]quinolizine-4-thiones 3 has been developed. The methodology involves ring annulation of 3,4-dihydro-6,7-dimethoxy-1 methylisoquinoline 1 with a variety of readily accessible acyclic and cyclic beta oxodithioesters 2 in the presence of triethylamine in refluxing benzene. These benzo[a]quinolizine-4-thiones can be readily converted to the corresponding benzo[a]quinolizine-4-ones 5 via dethiomethylative hydrolysis of the respective benzo[a]quinolizinium salts 4 obtained by alkylation of 3 with methyl iodide. PMID- 11430042 TI - Organic synthesis in ionic liquids: the Stille coupling. AB - [figure: see text] The Stille coupling reaction has been performed in 1-butyl-3 methylimidazolium tetrafluoroborate (BMIM BF4), a room-temperature ionic liquid (RTIL). Use of this solvent system allows for facile recycling of the solvent and catalyst system, which can be used at least five times with little loss in activity. An interesting preference in starting catalyst oxidation state for use with aryl bromides and aryl iodides was observed. PMID- 11430043 TI - Formal synthesis of (+)-3-deoxy-D-glycero-D-galacto-2-nonulosonic acid (KDN) via desymmetrization by ring-closing metathesis. AB - [figure: see text] Formal synthesis of the naturally occurring deaminated sialic acid KDN, a potential oncofetal antigen, has been accomplished in 45% overall yield via a novel ketalization/ring-closing metathesis sequence. The rapid introduction of all oxygen functionality in a completely stereocontrolled manner exploited a rigid 6,8-dioxabicyclo[3.2.1]oct-2-ene ring system. This general synthetic strategy should provide access to a number of KDN and sialic acid analogues. PMID- 11430044 TI - Catalytic ethenylation reaction of phenol using SnCl4. AB - [figure: see text] Ethenylation reaction of phenol with silylethyne at the o position is catalyzed by the SnCl4-BuLi reagent. While turn over numbers (TONs) in the reactions of m- or p-substituted phenols are 3 to 4, those for o substituted phenols are 8 to 9. PMID- 11430045 TI - General synthetic route to chiral flexible biphenylphosphine ligands: the use of a chiral additive enables the preparation and observation of metal complexes incorporating the enantiopure form. AB - [figure: see text] The enantio- and diastereomerically pure metal complex of a chirally flexible BIPHEP ligand is obtained through enantiomer-selective coordination of a BIPHEP-Ru complex with enantiopure 3,3' dimethyldiaminobinaphthyl, DM-DBN, followed by epimerization of the remaining BIPHEP-Ru enantiomer to complex with DM-DABN. Thus, an efficient and general synthetic route to a variety of substituted BIPHEP ligands from biphenol and observation of the enantiomerically pure BIPHEP ligands in their Ru(II) complexes are described. PMID- 11430046 TI - Candelalides A-C: novel diterpenoid pyrones from fermentations of Sesquicillium candelabrum as blockers of the voltage-gated potassium channel Kv1.3. AB - [figure: see text] Blockers of the voltage-gated potassium channel Kv1.3 are potential immunosuppressants. Candelalides A-C are three novel diterpenoid pyrones that block this channel. The structure, stereochemistry, and activity against Kv1.3 are described. PMID- 11430047 TI - The first total synthesis of (-)-solanapyrone E based on domino Michael strategy. AB - [figure: see text] A phytotoxin, solanapyrone E, has been synthesized from the decalone prepared by the domino Michael reaction of the kinetic enolate of optically pure acetylcyclohexene with methyl crotonate. After several transformations on the decalone ring, condensation of a methyl acetoacetate equivalent installed a pyrone moiety and introduction of a hydroxymethyl unit into the pyrone ring furnished solanapyrone E. PMID- 11430048 TI - Reductive ring opening of o-nitrobenzylidene acetals of monosaccharides: synthesis and photolysis of some photolabile sugars. AB - [figure: see text] A 6-O-o-nitrobenzyl methylglucoside and methylmannoside were synthesized by reacting 4,6-O-o-nitrobenzylidene acetals with triethylsilane and boron trifluoride etherate. A 2,6-di-O-o-nitrobenzyl and a 3,6-di-O-o-nitrobenzyl methylmannoside were obtained from a 2,3:4,6-di-O-o-nitrobenzylidene methylmannoside by the same method. The photolabile sugars obtained were deprotected by irradiation at 350 nm to afford methylglycosides. PMID- 11430049 TI - Synthesis of the left-hand portion of geldanamycin using an anti glycolate aldol reaction. AB - [figure: see text] A synthesis of the left-hand portion of the ansamycin antitumor natural product geldanamycin is reported. An advanced intermediate incorporates the methoxyquinone precursor as a pentasubstituted benzene with a 10 carbon chain that contains 4 of the 6 stereocenters. The key reaction is a novel anti glycolate aldol reaction with a new diaryl-4-oxapyrone used to generate the C-11, C-12 hydroxy, methoxy functionality. PMID- 11430050 TI - The first total synthesis of (+/-)-eremopetasidione. AB - [figure: see text] The total synthesis of racemic eremopetasidione, a norsesquiterpenoid, has been achieved in nine steps and 30% overall yield starting from creosol (5). Diels-Alder reaction of masked o-benzoquinone 6 and ethyl vinyl ketone and Cope rearrangement of 2-silyloxy-1,5-dienone 3 are the key steps. PMID- 11430051 TI - Selective binding of cis-1,3,5-cyclohexane tricarboxylic acid vs its epimeric trans isomer by a tripodal amidopyridine receptor; crystal structures of the 1:1 complexes. AB - [figure: see text] A tripodal tris-amidopyridine receptor forms a 1:1 complex with trans-1,3,5-cyclohexane tricarboxylic acid that is 1 order of magnitude less stable than the one formed with the corresponding cis-triacid epimer. The X-ray crystal structures of the complexes have been determined, confirming the binding geometry derived from NMR data in solution and force-field calculations, and its geometrical features are used to explain the observed selectivity. PMID- 11430052 TI - Oxazole synthesis with minimal purification: synthesis and application of a ROMPgel Tosmic reagent. AB - [figure: see text] The synthesis of ring opening metathesis, polymer-supported Tosmic reagent 1 is described. This reagent was utilized in the conversion of aldehydes to oxazoles in good yields and purities. PMID- 11430053 TI - Synthesis of 5'-deoxy-5'-thioguanosine-5'-monophosphorothioate and its incorporation into RNA 5'-termini. AB - [figure: see text] 5'-Deoxy-5'-thioguanosine-5'-monophosphorothioate (GSMP) was synthesized in four steps with 35% overall yield. GSMP serves as a good substrate for in vitro transcription with T7 RNA polymerase to yield 5'-GSMP-RNA, which was converted to 5'-HS-RNA by dephosphorylation with alkaline phosphatase. The thiol reactive agents can be efficiently introduced into the 5'-terminus of RNA. PMID- 11430054 TI - Synthetic studies toward anisatin: a formal synthesis of (+/-)-8-deoxyanisatin. AB - [figure: see text] An efficient strategy to construct the congested C-7a quaternary chiral center of anisatin was developed, by way of an Eschenmoser Claisen rearrangement. Conversion of the resultant amide to Kende's epsilon lactone intermediate 3 in four steps completed a concise formal synthesis of (+/ )-8-deoxyanisatin (2). PMID- 11430056 TI - Asymmetric autocatalysis of a pyrimidyl alkanol induced by chiral monosubstituted [2.2]paracyclophanes. AB - [figure: see text] Monosubstituted[2.2]paracyclophanes 3a-c with planar chirality were found to act as chiral initiators in an enantioselective addition of diisopropylzinc to 2-alkynylpyrimidine-5-carbaldehyde 1 to afford 2 alkynylpyrimidyl alkanol 2 with up to 97% ee. PMID- 11430055 TI - A thioesterase for chemoselective hydrolysis of S-acyl sulfanylalkanoates. AB - [figure: see text] A thioesterase, isolated from a strain of Alcaligenes sp. ISH108, chemoselectively hydrolyzes thiol esters. The application of the enzyme has been demonstrated in the preparation of the antihypertensive agent captopril. PMID- 11430057 TI - Enantio- and diastereocontrolled synthesis of an angular triquinane sesquiterpene (+)-arnicenone. AB - [figure: see text] (+)-Arnicenone, a sesquiterpene of an angular triquinane isolated from Arnica plants, has been synthesized for the first time in an enantiocontrolled manner from a synthetic equivalent of chiral 2 hydroxymethylcyclopentadienone to determine the absolute configuration of the natural product as 1R,3aR,5aS,8aR. PMID- 11430058 TI - Palladium-catalyzed regioselective arylation of an electron-rich olefin by aryl halides in ionic liquids. AB - [figure: see text] Palladium-catalyzed arylation of the electron-rich olefin butyl vinyl ether has been accomplished in the ionic liquid 1-butyl-3 methylimidazolium tetrafluoroborate ([bmim][BF4]), using as the arylating agents aryl iodides and bromides instead of the commonly used, but commercially unavailable and expensive, aryl triflates. The reaction proceeds with high efficiency and remarkable regioselectivity, leading almost exclusively to substitution by various aryl groups at the olefinic carbon alpha to the heteroatom of butyl vinyl ether. PMID- 11430059 TI - Unprecedented effects of achiral oxazolidinones on enantioselective radical mediated conjugate additions using a chiral zinc triflate. AB - [figure: see text] A role of achiral oxazolidinones to enhance the enantioselectivity in reactions of N-cinnamoyloxazolidinones with alkyl radicals promoted by a chiral Lewis acid is described. Efficient enantioselective radical mediated conjugate additions of N-cinnamoyloxazolidinone can be realized by use of a chiral zinc triflate generated from a readily prepared chiral bisoxazoline and an achiral oxazolidione. The NH moiety of achiral oxazolidinones is found to be necessary to enhance the enantioselectivity. PMID- 11430060 TI - Stereoselective synthesis of 2-hydroxy-alpha-mannopyranosides from glucal donors. AB - [figure: see text] Direct synthetic access to 2-hydroxy-alpha-mannopyranosides from glucal donors is accomplished via a one-pot stereoselective oxidative glycosylation reaction, employing the reagent combination of dibenzothiophene bis(triflate) and dibenzothiophene-5-oxide. PMID- 11430061 TI - A solid phase reagent for the capture phosphorylation of carbohydrates and nucleosides. AB - [figure: see text] A 1% cross-linked divinylbenzene-polystyrene copolymer, containing cyanoethoxy N,N-diisopropylamine phosphine was prepared as a phosphitylating agent. The polymer-bound phosphitylated precursor was subjected to reaction with alcohols in the presence of 1H-tetrazole to produce the corresponding polymer-bound phosphite triesters. These were then oxidized with tert-butyl hydroperoxide to give the polymer-bound monophosphate triesters. Removal of cyanoethoxy on the resin with 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) followed by basic cleavage of the p-hydroxybenzyl linker products yielded monophosphate derivatives. PMID- 11430062 TI - Rhodium-catalyzed Beckmann rearrangement. AB - [figure: see text] Beckmann rearrangement of oxime is catalyzed by [RhCl(cod)]2, trifluoromethanesulfonic acid, and tris(p-tolyl)phosphine in refluxing dichloroethane, giving the corresponding amide in good yield. Product/acid ratios of 10:20 can be attained in the reaction of benzophenone oximes. PMID- 11430063 TI - Metal-mediated self-assembly of pyridylcalixarenes: prevention of intramolecular metal chelation is essential in constructing molecular capsules. AB - To construct calixarene-based molecular capsules utilizing the pyridyl-Pd(II) interaction, reactions of cone-pyridylcalix[4]arene 3, cone-pyridylcalix[5]arene 13, and cone-pyridylcalix[4]arene bis-crown 16 with square-planar Pd(II) complex 7 were investigated. Because of the coexistence of intermolecular binding and chelate-forming intramolecular binding, the reactions of cone pyridylcalix[4]arene 3 or cone-pyridylcalix[5]arene 13 with cis-Pd(II) complex 7 yield complicated, structure-unknown oligomers. The short dioxyethylene bridges on the lower rim of pyridylcalix[4]arene bis-crown 16 rigidify the cone conformation and thus prohibit 16 from the intramolecular binding with a metal component. Thus, two cone-tetrapyridylcalix[4]arene bis-crown 16 and four cis Pd(II) complex molecules self-assemble into molecular capsules that exist as a parallel/antiparallel conformer mixture in a nearly 1:1 ratio. The results demonstrated that to prevent entropically favorable intramolecular binding is essential is constructing higher capsule-like structures with calixarene building blocks by self-assembling. PMID- 11430065 TI - Enantioselective synthesis of optically active carbocyclic sugars. PMID- 11430064 TI - Catalytic, highly enantioselective Friedel-Crafts reactions of aromatic and heteroaromatic compounds to trifluoropyruvate. A simple approach for the formation of optically active aromatic and heteroaromatic hydroxy trifluoromethyl esters. AB - A new catalytic enantioselective synthetic method for the formation of optically active aromatic and heteroaromatic hydroxy-trifluoromethyl ethyl esters is presented. This catalytic enantioselective Friedel-Crafts reaction of trifluoromethyl pyruvate with aromatic and heteroaromatic compounds is catalyzed by a chiral bisoxazoline copper(II) complex and proceeds in good yield and with high enantiomeric excess. For a series of substituted indoles, the corresponding 3-substituted hydroxy-trifluoromethyl ethyl esters are formed in up to 93% yield and 94% ee. Pyrrole and 2-substituted pyrroles also react with trifluoromethyl pyruvate in a highly enantioselective aromatic electrophilic reaction and up to 93% ee and good yields are obtained. Furanes and thiophenes give the corresponding 2-hydroxy-trifluoromethyl ethyl esters in high enantiomeric excess; however, the yields of the products are only moderate. Various types of aromatic compounds react in this catalytic reaction with trifluoromethyl pyruvate to give the aromatic electrophilic addition product in good yield. To obtain high enantiomeric excess (> 80% ee) it is necessary that aromatic amines are protected with sterically demanding protecting groups such as benzyl or allyl. This prevents coordination of the amine nitrogen atom to the catalyst, as aromatic amines having a N,N-dimethyl group probably coordinate to the catalyst, leading to a significant reduction of the enantioselective properties of the catalyst. On the basis of the experimental results and the absolute configuration of the formed chiral center, the mechanism for the catalytic enantioselective Friedel Crafts reaction is discussed. PMID- 11430066 TI - Synthesis of homochiral 5- and 8-substituted 2-[((2-(2,6-dimethoxyphenoxy) ethyl)amino)methyl]-1,4-benzodioxanes and electrophoretic determination of their enantiomeric excess. PMID- 11430067 TI - A trial toward the synthesis of gamma-fluorinated beta-amino-alpha thiohydroxyamino acid: a finding of desulfurizative C-C bond formation in thio Wittig-type rearrangement of S-(N-aryl-trifluoroacetimidoyl)thioglycolates. PMID- 11430068 TI - A general solution- and solid-phase synthetic procedure for incorporating three contiguous imidazole moieties into DNA sequence reading polyamides. PMID- 11430069 TI - Efficient carbamate synthesis via a three-component coupling of an amine, CO2, and alkyl halides in the presence of Cs2CO3 and tetrabutylammonium iodide. PMID- 11430070 TI - Practical and efficient synthesis of orthogonally protected constrained 4 guanidinoprolines. PMID- 11430071 TI - From amides to amidines: preparations of imidoylbenzotriazoles and arylaminoheterocycles. PMID- 11430072 TI - Cp2TiCl-mediated selective reduction of alpha,beta-epoxy ketones. PMID- 11430073 TI - Total syntheses of pyranoquinoline alkaloids: simulenoline, huajiaosimuline, and (+/-)-7-demethoxyzanthodioline. PMID- 11430074 TI - Mixed phosphinine-ether macrocycles. PMID- 11430075 TI - A practical synthesis of trans-dichlororuthenium ((S,S)-2,6-bis(4-isopropyl-2 oxazolin-2-yl)-pyridine)(ethylene) amenable to large-scale preparation. PMID- 11430076 TI - 2-trifluoromethoxyethyl triflate: a versatile trifluoromethoxyethyl carrier. PMID- 11430077 TI - Chemistry of trichlorofluoromethane: synthesis of chlorofluoromethyl phenyl sulfone and fluoromethyl phenyl sulfone and some of their reactions. AB - It was observed that the reaction of CFCl3 with thiophenoxide gave only 10% of the corresponding thioether. On the other hand, these thioethers could be prepared in excellent yield from diaryl disulfides and CFCl3 in the presence of sodium hydroxymethanesulfinate in aqueous DMF at 4 atm pressure of nitrogen. Dechlorination of the thioether (PhSCFCl2) with different reducing agents were studied. Most of the reducing agents eliminated both fluorine and chlorine functionalities or gave the hydrolyzed products. But its sulfone on treatment with Zinc in methanol gave monochlorofluoromethyl and fluoromethyl phenyl sulfone in good yields. Darzens reaction of these compounds was also studied. PMID- 11430078 TI - Synthesis of analogues of Eunicea gamma-cembranolides containing cyclic ethers via saponification. AB - A method for the synthesis of derivatives of the lead structures euniolide (1), 12,13-bisepieupalmerin (2), and eupalmerin acetate (3) containing tetrahydrofuran and tetrahydropyran ring systems was developed on the basis of alkali-induced intramolecular oxacyclizations. Representatives of the new analogues were submitted to the in vitro antitumor cell-line-screening program of the National Cancer Institute (NCI). While it was shown that a variety of structural modifications are possible, these transformations led typically to nontoxic synthetic cembranoids. PMID- 11430079 TI - Photochemistry and photophysics of triarylmethane dye leuconitriles. AB - The photochemical reactions of crystal violet leuconitrile (CVCN) were investigated by the means of product analysis and trapping experiments, laser flash and steady-state photolysis, and steady-state fluorescence. The influence of oxygen on the reaction was examined in detail. The photochemistry of malachite green leuconitrile (MGCN), basic fuchsin leuconitrile (BFCN), and crystal violet leucomethyl (CVMe) and leucobenzyl (CVBn), as well as triphenylacetonitrile, was studied. The results suggest ionization occurs from S1, while the di-pi-methane reaction is the photochemical route from T1. PMID- 11430080 TI - A density functional theory study on the mechanism of the permanganate oxidation of substituted alkenes. AB - Oxidation of substituted alkenes by permanganate follows a (3 + 2) cycloaddition mechanism. DFT calculations (Becke3LYP/6-31G(d)) strongly favor the (3 + 2) pathway against a (2 + 2) pathway that proceeds through a metallaoxetane. The difference in free activation energy between the two pathways is around 40 to 45 kcal/mol for the nine compounds covered. The results for the (3 + 2) cycloaddition mechanism are in agreement with experimentally observed kinetic data reported earlier. Symmetric transition states are calculated for alkenes with at least one CH2 group between the double bond and the acid group, while all others are unsymmetrical due to the repulsion between the permanganate oxygens and the carboxylic oxygens. Steric bulk, introduced by the number and position of substituents at the double bond, has no significant effect on the activation energies. A higher level of theory (Becke3LYP/6-311G(d,p)) leads to a reduction of the rmsd between experimental and calculated values, but has little influence on the calculated geometries. PMID- 11430081 TI - Reaction of primary aromatic amines with alkyl carbonates over NaY faujasite: a convenient and selective access to mono-N-alkyl anilines. AB - At atmospheric pressure and at 130-160 degrees C, primary aromatic amines (p XC6H4NH2, X = H, Cl, NO2) are mono-N-alkylated in a single step, with symmetrical and asymmetrical dialkyl carbonates [ROCOOR', R = Me, R' = MeO(CH2)2O(CH2)2; R = R' = Et; R = R' = benzyl; R = R' = allyl; R = Et, R' = MeO(CH2)2O(CH2)2], in the presence of a commercially available NaY faujasite. No solvents are required. Mono-N-alkyl anilines are obtained with a very high selectivity (90-97%), in good to excellent yields (68-94%), on a preparative scale. In the presence of triglyme as a solvent, the mono-N-alkyl selectivity is independent of concentration and polarity factors. The reaction probably takes place within the polar zeolite cavities, and through the combined effect of the dual acid-base properties of the catalyst. PMID- 11430082 TI - Photochemical E (trans)-->Z (cis) isomerization in substituted 1 naphthylacrylates. AB - Substituted naphthylacrylates, 1-3, not showing rotamerism have been synthesized with a view to study photochemical E (trans)-->Z (cis) isomerization. Photostationary state composition of the isomers upon direct excitation, triplet sensitized isomerization, quantum yield of isomerization, and steady state and time-resolved fluorescence behavior have been studied for these naphthylacrylates. The direct excitations of the compounds yield high Z (approximately 80%) isomer composition, whereas the triplet sensitization results in less Z (approximately 20%) isomer composition. This indicates that the singlet pathway is very efficient in converting the E isomer to the Z isomer. The naphthylacrylates 1 and 2 exhibit structured fluorescence at room temperature in hexane and upon changing the solvent to CH3CN; the structure of the fluorescence is lost, indicating that the singlet excited-state develops a polar character in a polar environment. The polar nature of the singlet excited state becomes more clear in the case of 3 from its fluorescence solvatochromism. The naphthylacrylates did not exhibit excitation wavelength-dependent fluorescence at room temperature suggesting that the ground state conformers (rotamers) are not involved. Fluorescence lifetimes measured for these compounds displayed biexponential behavior, which is explained using a two-state model. PMID- 11430083 TI - alpha-, beta-, and gamma-Cyclodextrin dimers. Molecular modeling studies by molecular mechanics and molecular dynamics simulations. AB - The alpha-, beta-, and gamma-cyclodextrin (CyD) dimers have been studied by molecular mechanics (MM) and molecular dynamics (MD) calculations, and the relative stability of dimers and the involved molecular interactions have been determined. Three possible orientations were considered for the alpha-, beta-, and gamma-CyD dimers: the head-to-head, the head-to-tail, and the tail-to-tail. In vacuo MM calculations were used to obtain the most stable arrangements, and MD simulations were performed over all energy minima obtained for each dimer. Results from MD always show head-to-head orientation as the most stable as a result of the larger number of intermolecular hydrogen bonds present. PMID- 11430084 TI - The particular sensitivity of silyl ethers of D-glucal toward two Vilsmeier-Haack reagents POCl3.DMF and (CF3SO2)2O.DMF. Their unique and selective conversion to the corresponding C(6)-O-formates. AB - The two electrophilic Vilsmeier-Haack reagents POCl3.DMF 2 or (CF3SO2)2O.DMF 3 mediate the one-step and selective conversion of O-triethylsilyl (O-TES), O-tert butyldimethylsilyl (O-TBDMS), O-tert-butyldiphenylsilyl (O-TBDPS), and O triisopropylsilyl (O-TIPS) ethers of D-glucal to the corresponding C(6)-O formates. PMID- 11430085 TI - Small ring constrained peptidomimetics. Synthesis of epoxy peptidomimetics, inhibitors of cysteine proteases. AB - Different dipeptide analogues containing an oxirane ring in the place of the peptidic bond were prepared starting from naturally occurring amino acids. N-Fmoc amino aldehydes were transformed into the corresponding methoxyvinyl derivatives through a Wittig reaction, and the addition of PhSeCl gave a series of different alpha-phenylselenyl aldehydes. Mukajiama reaction with silylketene acetals gave an intermediate product that was finally transformed into the desired oxiranyl peptidomimetics. Following this strategy we were able to control three new contiguous stereocenters starting from the enantiomerically pure amino acid. The dipeptide analogues could be used in SPPS on a SASRIN resin as the final epoxides were relatively unstable under acidic conditions. Moreover the synthesis of the single dipeptide mimetics was carried out on solid phase to generate a small library of epoxy peptidomimetics. Some of the products prepared in this work resulted as time-dependent reversible inhibitors of cysteine protease. PMID- 11430086 TI - Synthesis of pyrrolidinone PNA: a novel conformationally restricted PNA analogue. AB - To preorganize PNA for duplex formation, a new cyclic pyrrolidinone PNA analogue has been designed. In this analogue the aminoethylglycine backbone and the methylenecarbonyl linker are connected, introducing two chiral centers compared to PNA. The four stereoisomers of the adenine analogue were synthesized, and the hybridization properties of PNA decamers containing one analogue were measured against complementary DNA, RNA, and PNA strands. The (3S,5R) isomer was shown to have the highest affinity toward RNA, and to recognize RNA and PNA better than DNA. The (3S,5R) isomer was used to prepare a fully modified decamer which bound to rU10 with only a small decrease in Tm (delta Tm/mod = 1 degree C) relative to aminoethylglycine PNA. PMID- 11430087 TI - Molecular saddles. 4. Redox-active cyclophanes by bridging the 9,10-bis(1,3 dithiol-2-ylidene)-9,10-dihydroanthracene system: synthesis, electrochemistry, and X-ray crystal structures of neutral species and a dication salt. AB - We report the synthesis of a new series of cyclophanes 11a-d by ester-forming macrocyclization reactions of diol 9 with the dicarbonyl chloride derivatives of benzene, thiophene, ferrocene, and diphenyl ether, 10a-d, respectively. Compounds 11a-d display a two-electron, quasireversible oxidation wave in the cyclic voltammogram to yield the dication species at Eoxpa = 0.70-0.72 V (for 11a-c) and 0.47 V (for 11d) (vs Ag/AgCl in acetonitrile). The raised oxidation potentials for 11a-c reflect the reduced stability of the twisted dication structure within the steric constraints of the smaller cyclophanes. Consistent with this, the value of delta E (defined as Eoxpa - Eoxpc) decreases (i.e., reversibility of the oxidation process increases) in the sequence 11d > 11c > 11a > 11b as the bridging chain becomes shorter. X-ray crystal structures are reported for compounds 11a-d and the dication salt 11d2+(I3-)2.(CH2Cl2)2.25. For 11a-d the typical saddle-shaped conformation of the 9,10-bis(1,3-dithiol-2-ylidene)-9,10 dihydroanthracene moiety is observed, with the strain imposed by the cyclophane ring being accommodated in the structure of the bridging unit. In the dication 11d2+ both dithiolium rings are planar and the anthracene unit is essentially aromatic, with the conformation of the bridge basically the same as in neutral 11d. PMID- 11430088 TI - Bis(1,3,4-thiadiazolo)-1,3,5-triazinium halides. 2. Intramolecular ring transformation and synthesis of novel highly substituted guanidines. AB - Bis(1,3,4-thiadiazolo)-1,3,5-triazinium halides 6 can be easily attacked by nucleophiles at either the C(3a) or the C(4a) position of the central six membered (cationic) ring. Nucleophilic attack leads to at least two reaction channels, one of which has been previously detected (pathway a) and leads to novel aminals 19. In this paper we report on a second channel (pathway b). Attack of primary or secondary amines 8 at C(3a) or C(4a) in 6 (and their analogues 7) leads to the weakly stabilized intermediates 14. A cascade of several proton shifts, ring openings, rearrangements, and ring closure processes is initiated which finally leads via 17 and 18 to novel highly substituted guanidines 9, 10, 12, and 13. Pathway b seems to be the result of well-balanced negative hyperconjugative effects in 14 and/or 17 which control the highly selective opening of a relatively stable central 1,3,5-triazinium ring to yield the crucial intermediate 18. Some representatives of the guanidines have been characterized by X-ray analyses. Since some of the guanidines contain one or two chirality centers, an effort was made to investigate the stereochemistry of these compounds. PMID- 11430089 TI - Synthesis of oligomers of trans-(4S,5R)-4-carboxybenzyl 5-methyl oxazolidin-2 one: an approach to new foldamers. AB - The synthesis of two oligomers containing three and four residues, respectively, of trans-(4S,5R)-4-carboxy 5-methyloxazolidin-2-ones is described. The monomer is obtained by starting from benzyl-N-Boc-(3R)-aminobutanoate, by cyclization into the corresponding trans-(2S,3R)-2-carboxybenzyl-3-methyl-N-Boc-aziridine and rearrangement of the product to trans-(4S,5R)-4-carboxybenzyl-5- methyloxazolidin 2-one, catalyzed by Sn(OTf)2. The oligomers are synthesized by activating the carboxy group as its pentaflourophenyl ester. The trimer and the tetramer are obtained in good yield, and their 1H NMR spectra suggest that these molecules fold in ordered structures, where the C-4 hydrogen of a ring is always close to the carbonyl of the next ring. This result shows that the 4-carboxy-5-substituted oxazolidin-2-ones are a new class of pseudoprolines which fully control the formation of a Xaai-1-Proi peptide bond in the trans conformation and are complementary to the pseudoprolines obtained from cyclocondensation of cysteine, serine, or threonine and aldehydes or ketones, which strongly favor the Xaai-1 Proi peptide bond in the cis conformation. PMID- 11430090 TI - Asymmetric Baeyer-Villiger oxidations of 4-mono- and 4,4-disubstituted cyclohexanones by whole cells of engineered Escherichia coli. AB - Whole cells of an Escherichia coli strain that overexpresses Acinetobacter sp. NCIB 9871 cyclohexanone monooxygenase have been used for the Baeyer-Villiger oxidations of a variety of 4-mono- and 4,4-disubstituted cyclohexanones. In cases where comparisons were possible, this new biocatalytic reagent provided lactones with chemical yields and optical purities that were comparable to those obtained from the purified enzyme or a strain of bakers' yeast that expresses the same enzyme. The efficient production of cyclohexanone monooxygenase in the E. coli expression system (ca. 30% of total soluble protein) allowed these oxidations to reach completion in approximately half the time required for the engineered bakers' yeast strain. Surprisingly, 4,4-disubstituted cyclohexanones were also accepted by the enzyme, and the enantioselectivities of these oxidations could be rationalized by considering the conformational energies of bound substrates along with the enzyme's intrinsic enantioselectivity. The enzyme expressed in E. coli cells also oxidized several 4-substituted cyclohexanones bearing polar substituents, often with high enantioselectivities. In the case of 4 iodocyclohexanone, the lactone was obtained in > 98% ee and its absolute configuration was assigned by X-ray crystallography. The crystal belongs to the monoclinic P2(1) space group with a = 5.7400(10), b = 6.1650(10), c = 11.377(2) A, b = 99.98(2) degrees, and Z = 2. Taken together, these results demonstrate the utility of an engineered bacterial strain in delivering useful chiral building blocks in an experimentally simple manner. PMID- 11430091 TI - Diastereoselective synthesis of polysubstituted tetrahydropyrans and thiacyclohexanes via indium trichloride mediated cyclizations. AB - Polysubstituted tetrahydropyrans and thiacyclohexanes were synthesized in high yields with excellent diastereoselectivities via indium trichloride mediated cyclizations between homoallyl alcohols and mercaptans with aldehydes. In the case of tetrahydropyran products, the stereochemistry of the product was found to be directly correlated with the geometry of the homoallyl alcohols; whereas the cross-cyclization of aldehydes with trans-homoallyl alcohols generated (up-down up) 2,3,4-trisubstituted tetrahydropyran products exclusively, the reaction of aldehydes with cis-homoallyl alcohols provided mainly (up-up-up) 2,3,4 trisubstituted products. When a trisubstituted homoallyl alcohol was used, its cross-cyclization with aldehydes generated (up-down-up-down-up) pentasubstituted tetrahydropyran derivatives with simultaneous controlling of five stereogenic centers. On the other hand, a cyclization-decyclization equilibrium was observed in the formation of thiacyclohexanes. The reaction of both cis- and trans homoallyl mercaptans with aldehydes provided the same major diastereomers. PMID- 11430092 TI - Conformational studies by dynamic NMR. 79. Dimesityl sulfine revisited: detection of the helical antipodes and determination of their enantiomerization pathways. AB - By means of low-temperature NMR spectra, it is demonstrated that dimesityl sulfine (Mes2C=SO) adopts in solution the same chiral propeller conformation (C1 symmetry) determined by X-ray diffraction in the crystalline state. With the help of MM calculations, it has been also shown that a correlated rotation (cog wheel effect) of the two mesityl rings reverses the molecular helicity according to an enantiomerization process entailing a one-ring flip pathway with delta G++ = 5.9 kcal mol-1 and a two-ring flip pathway with delta G++ = 13.8 kcal mol-1. On the contrary the Z- and E-isomers of mesityl phenyl sulfine (MesPhC=SO) adopt essentially achiral conformations (Cs symmetry), having the Ph-CSO rotation barriers equal to 5.2 and 5.8 kcal mol-1, respectively, and the mesityl-CSO rotation barriers equal to 21.3 and 15.1 kcal mol-1, respectively. PMID- 11430093 TI - Enhancing the dienic reactivity of phospholes: an improved access to trivalent 7 phosphanorbornenes. AB - A structural comparison of 1-cyano- and 1-alkoxy-3,4-dimethylphospholes with 1 benzylphosphole has led to some unexpected conclusions. There is no univocal relationship between phosphole aromaticity and pyramidality at phosphorus. It has been found that both the highly pyramidal 1-cyanophosphole 1 (sigma(CPC angles) = 290 degrees), and the much less pyramidal 1-alkoxyphosphole 6 (sigma(CPC angles) = 310 degrees) have a low Bird aromaticity index (27 for both molecules), when compared to 1-benzylphosphole (sigma(CPC angles) = 303 degrees, BI = 35.5). This low aromaticity is correlated with a high reactivity of the diene in both 1 and 7 (similar to 6) toward acrylonitrile. Good stereochemical control is observed with 7, which gives exclusively the anti,endo [4 + 2] cycloadducts with acrylonitrile and diethyl vinylphosphonate. PMID- 11430094 TI - Steric and electronic ligand perturbations in catalysis: asymmetric allylic substitution reactions using C2-symmetrical phosphorus-chiral (bi)ferrocenyl donors. AB - Three series of P-chiral diphosphines based on ferrocene (1a-f, 2a-c) and biferrocenyl skeletons (3a-c), including novel ligands 1f and 3c, were employed in palladium-catalyzed allylic substitution reactions. Steric effects imposed by the phosphine residues were studied using C2-symmetrical donors 1 (1 = 1,1' bis(arylphenylphosphino)ferrocene with aryl groups a = 1-naphthyl, b = 2 naphthyl, c = 2-anisyl, d = 2-biphenylyl, e = 9-phenanthryl, and f = ferrocenyl), whereas para-methoxy- and/or para-trifluoromethyl substitution of the phenyl moieties in 1a enabled investigation of ligand electronic effects applying ferrocenyl diphosphines 2a-c. Ligands 3 (3 = 2,2'-bis- (arylphenylphosphino)-1,1' biferrocenyls with aryl substituents a,c = 1-naphthyl (diastereomers) and b = 2 biphenylyl) allowed for comparison of backbone structure effects (bite angle variation) in catalysis. Linear and cyclic allylic acetates served as substrates in typical test reactions; upon attack of soft carbon and nitrogen nucleophiles on (E)-1,3-diphenylprop-2-ene-1-yl acetate the respective malonate, amine, or imide products were obtained in enantioselectivities of up to 99% ee. A crystal structure analysis of a palladium 1,3-diphenyl-eta 3-allyl complex incorporating ligand (S,S)-1a revealed a marked distortion of the allyl fragment, herewith defining the regioselectivity of nucleophile addition. PMID- 11430095 TI - Aspirin. An ab initio quantum-mechanical study of conformational preferences and of neighboring group interactions. AB - The potential energy surface of acetylsalicylic acid, aspirin, has been explored at the RHF/6-31G* and B3LYP/6-31G* levels, and single-point calculations were performed at levels up to B3LYP/6-311G**//B3LYP/6-31G*. All conformational isomers have been located, the thermochemical functions have been computed, and relative energies and free enthalpies were determined. The conformational space of aspirin is spanned by three internal coordinates, and these are the carboxylic acid C-O conformation (s-trans preferred by about 7 kcal/mol), the C-COOH conformation (Z preferred unless there are H-bonding opportunities), and the ester C-O conformation (s-trans preferred by about 4 kcal/mol). There are nine aspirin isomers since one of the conformers realizes hydrogen-bonding structure isomerism as well. Neighboring group interactions are discussed with reference to the intrinsic properties of benzoic acid and phenyl acetate. The intrinsic conformational preference energies for benzoic acid and phenyl acetate are not additive. The acid s-trans preference energies differ by as much as 9 kcal/mol depending on the Ph-COOH and ester conformations. Similarly, the E-preference energies about the Ph-COOH bond vary by as much as 6 kcal/mol depending on the ester conformation. The structural discussion suggests an overall ortho repulsion between the functional groups in all aspirin isomers including the intramolecularly hydrogen-bonded isomers. The isodesmic reaction between the most stable conformers of benzoic acid and phenyl acetate to form aspirin and benzene is found to be endothermic by 2.7 kcal/mol and provides compelling evidence for and a quantitative measure of ortho repulsion. The ortho repulsion of 2.7 kcal/mol is a lower limit, and the ortho repulsion can increase to as much as 6 kcal/mol in some aspirin isomers. PMID- 11430096 TI - Substituent effects and charge delocalization mode in chrysenium, benzo[c]phenanthrenium, and benzo[g]chrysenium cations: a stable ion and electrophilic substitution study. AB - The first series of persistent carbocations derived from mono- and disubstituted chrysenes Ch (5- methyl- 3, 2-methoxy- 19, 2-methoxy-11-methyl- 20, 2-methoxy-5 methyl- 21, and 9-methyl-4H-cyclopenta[def]chrysene 22), monosubstituted benzo[c]phenanthrenes BcPh (3-methoxy- 23, 3-hydroxy- 24), and monosubstituted benzo[g]chrysenes BgCh (12-methoxy- 25; 12-hydroxy- 26) were generated in FSO3H/SO2ClF or FSO3H-SbF5 (4:1)/SO2ClF and studied by low-temperature NMR at 500 MHz. The methoxy and methyl substituents direct the protonation to their respective ortho positions. Whereas parent Ch 1 is protonated at C-6/C-12, 3 is protonated at C-6 (3aH+) and at C-12 (3bH+) with the latter being the thermodynamic cation. The 2-methoxy-Ch 19 is protonated at C-1 to give two conformationally distinct carboxonium ions (19aH+/19bH+). In the disubstituted Ch derivatives 20 and 21, the 2-methoxy overrides the 5-methyl and the predominant carbocations formed are via attack ortho to methoxy. For the methano derivative 22 (Me at C-9), a 3:1 mixture of 22aH+/22bH+ is formed. For parent BcPh 13, nitration and benzoylation are directed to C-5. With 3-methoxy-BcPh 23, the site of attack moves to C-4, thus producing two conformationally distinct carboxonium ions (23aH+/23bH+), whereas conventional nitration gave a 2:1 mixture of 23aNO2 and 23bNO2. In 3-hydroxy-BcPh 24, the carboxonium ion 24H+ is exclusively formed. For parent BgCh 16, protonation, nitration, and benzoylation are all directed to C-10 (16H+, 16NO2, 16COPh), but presence of OMe or OH substituent at C-12 changes the site of attack to C-11. Charge delocalization mode is probed based on magnitude of delta delta 13Cs and conformational aspects via NOED experiments. Complete NMR data are also reported for several benzoylation/nitration products. Using ab initio/GIAO (and NICS), the NMR chemical shifts (and aromaticity) in model carbocations A-D were evaluated. This work represents the first direct study of the carbocations derived from the methyl-, methoxy-/hydroxy-derivatives of three important classes of bay-region and fjord-region PAHs whose diol epoxides extensively bind to DNA. It also extends the available data on electrophilic chemistry of BcPh and BgCh. PMID- 11430097 TI - Radical and non-radical mechanisms for alkane oxidations by hydrogen peroxide trifluoroacetic acid. AB - The oxidation of cyclohexane by the H2O2-trifluoroacetic acid system is revisited. Consistent with a previous report (Deno, N.; Messer, L. A. Chem. Comm. 1976, 1051), cyclohexanol forms initially but then esterifies to cyclohexyl trifluoroacetate. Small amounts of trans-1,2-cyclohexadiyl bis-(trifluoroacetate) also form. Although these products form irrespective of the presence or absence of O2, dual mechanisms are shown to operate. In the absence of O2, the dominant mechanism is a radical chain reaction that is propagated by CF3. abstracting H from C6H12 and SH2 displacement of C6H11. on CF3CO2OH. The intermediacy of C6H11. and CF3. is inferred from production of CHF3 and CO2 along with cyclohexyl trifluoroacetate, or CDF3 when cyclohexane-d12 is used. In the presence of O2, fluoroform and CO2 are suppressed, the reaction rate slows, and the rate law approaches second order (first order in peracid and in C6H12). Trapping of cyclohexyl radicals by quinoxaline is inefficient except at elevated (approximately 75 degrees C) temperatures. Fluoroform and CO2, telltale evidence for the chain pathway, were not produced when quinoxaline was present in room temperature reactions. These observations suggest that a parallel, nonfree radical, oxenoid insertion mechanism dominates when O2 is present. A pathway is discussed in which a biradicaloid-zwiterionic transition state is attained by hydrogen transfer from alkane to peroxide oxygen with synchronous O-O bond scission. PMID- 11430098 TI - Highly regioselective cyclotrimerization of 1-perfluoroalkylenynes catalyzed by nickel. AB - We report the highly regioselective cyclotrimerization of the 1 perfluoroalkylenynes in the presence of Ni(PPh3)4. The substituent effects on the reactivity of the enynes were investigated. We also succeeded in the nickel catalyzed cocyclization of the 1-perfluoroalkylenynes with (trimethylsilyl) acetylene. The possible structures of the intermediates of the cyclotrimerization and the reasons for the observed high regioselectivity were discussed. PMID- 11430099 TI - Solid-state solvolysis of thiophene-substituted trityl-type alcohols: nucleophilic substitution induced by gas-solid contact. AB - The nucleophilic substitution reaction by gas-solid contact has been investigated. When 9-thienothienylfluoren-9-ol derivatives were coground with dichlorodicyanoquinone (DDQ) and then exposed to methanol vapor, the corresponding 9-methoxyfluorenes were obtained in 15-70% yields. Throughout the whole procedure the solid state was retained. The generation of a radical cation in the coground solids via charge-transfer interaction between the substrate alcohol and DDQ was suggested by the ESR spectrum. The mechanism involving the collapse of the radical cation to generate a proton, which acts as a catalyst to afford the carbocation, was deduced based on the electrochemical oxidation of the substrate in solution. The propagation of the substitution reaction in the solid state has been shown for the carbocation upon contact with methanol vapor. The crystalline inclusion compounds of 9-thienothienylfluoren-9-ol derivatives incorporating methanol as a guest were exposed to HCl gas. This gas-solid reaction also led to the formation of the corresponding methoxy compounds maintaining the solid state. Through this work a new consequence of solid-state cogrinding is deduced. PMID- 11430100 TI - Efficient synthesis of piperidine derivatives. Development of metal triflate catalyzed diastereoselective nucleophilic substitution reactions of 2-methoxy- and 2-acyloxypiperidines. AB - Nucleophilic substitution reactions of 2-methoxy- and 2-acyloxypiperidines were investigated. First, new and efficient methods for the preparation of the starting piperidine derivatives were developed. N-Benzyloxycarbonyl-2 methoxypiperidine (3) and 3-substituted-2-acyloxy-N-benzyloxycarbonylpiperidines (4a-d), which are recognized as the simplest imino-sugars, were prepared and were examined as substrates for nucleophilic substitution reactions with silyl enolates under the influence of catalytic amounts of metal triflates (Sc(OTf)3, Sn(OTf)2, Cu(OTf)2, Hf(OTf)4, etc). Among the triflates tested, Sc(OTf)3 gave the best results. It was found that 2-acetoxy-3-benzyloxy-N benzyloxycarbonylpiperidine (4a) reacted with silyl enolates to afford the 2 alkylated adducts in high cis-selectivity, while 2,3-diacyloxy-N benzyloxycarbonylpiperidines (4b-d) showed transselectivity. The stereochemical assignments were carefully performed using NMR analysis, X-ray crystallography, and synthetic transformations. Febrifugine (1), a potent antimalarial alkaloid, was successfully synthesized from 2,3-diacetoxy-N-benzyloxycarbonylpiperidine (4b) on the basis of these diastereoselective nucleophilic substitution reactions. PMID- 11430101 TI - A building block method for the synthesis of higher cycloamides. AB - Series of cyclic amides containing optically active helicene, (P)-1,12 dimethylbenzo[c]phenanthrene, are synthesized using a building block method. The building block consists of one (P)-helicene unit and one dianiline unit with its amino-terminal-protected with benzyloxycarbonyl and its acid terminal activated as acid chloride. The coupling with (P,P....)-[(n - 3) + (n - 2)]diamine followed by deprotection gives (P,P....)-[(n - 1) + n]diamine, which possesses n - 1 parts of (P)-helicene and n parts of dianiline. Cyclization of the (P,P....)-[(n - 1) + n]diamine with helicenediacid dichloride gives (P,P....)-[n + n]cycloamide. All the members of (P,P)-[2 + 2]cycloamide to (P,P,P,P,P,P,P,P,P,P)-[10 + 10]cycloamide are synthesized using this method, and are compared spectroscopically. PMID- 11430102 TI - Evaluation of the extent of conjugation in symmetrical and asymmetrical aryl substituted acetophenone azines using electrochemical methods. AB - The electrochemical behavior of a series of symmetrical and unsymmetrical aryl substituted acetophenone azines (1-X/Y, where X and Y are 4-NO2, 4-CN, H, 3-OCH3, 4-OCH3, 4-CH3, and 4-N(CH3)2) was studied in acetonitrile and N,N dimethylformamide (DMF) solution using cyclic voltammetry (CV). Compounds 1-X/Y, where neither X or Y are nitro substituents, undergo successive reduction to their radical anion (1-X/Y.-) and then dianion (1-X/Y2-), respectively. In all cases, the formation of the radical anion is completely reversible and the standard reduction potentials, Eo1-X/Y/1-X/Y.- could be determined. The reversibility of the second electron transfer is substituent dependent with certain dianions sufficiently basic to be protonated under our conditions. Standard reduction potentials (Eo1-X/Y/1-X/Y.-) for the formation of radical anions exhibit a large substituent effect with values differing by more than 0.66 V throughout the series going from 1-4-CN/4-CN to 1-4-OCH3/4-OCH3; similar substituent effects were determined for the formation of the dianion. The nitro containing azines deviate from the above-mentioned behavior. With the exception of 1-4-NO2/4-NO2, they exhibit single electron waves that have values of Eo1 X/Y/1-X/Y.- within 40 mV of each other and thus the reduction is not subject to the same substituent effect as the other azines. 1-4-NO2/4-NO2 exhibits an Eo at a similar potential, but is a two-electron reversible wave with features indicative of a reduction system containing two localized, nonconjugated redox centers. The reduction potentials of all the aryl azines were correlated with Hammett sigma parameters to look at variations in Eo1-X/Y/1-X/Y.- vs SCE as a function of substituent. The small rho values in combination with the other electrochemical data provide support for single bond character of the N-N bond and evidence for a lack of strong electronic communication between the two aryl centers through the azomethine bonds, especially for those systems with electron withdrawing groups. PMID- 11430103 TI - Trapping of the putative cationic intermediate in the Morin rearrangement with carbon nucleophiles. AB - This paper presents reactions in which the putative cationic intermediate in the Morin rearrangement is trapped by aromatic carbon nucleophiles (indoles and furans). For example, reaction of sulfoxide 27 with trifluoroacetic acid in chloroform provides, among other products, indole 29 and indoline 30. The indoline was shown to be in equilibrium with the nine-membered ring bridged indole 31. Other examples of Morin rearrangement-trapping reactions are presented, and mechanisms for these transformations are proposed. PMID- 11430104 TI - Enantioselective syntheses of monotetrahydrofuran Annonaceous acetogenins tonkinecin and annonacin starting from carbohydrates. AB - The total synthesis of two mono-THF acetogenins, tonkinecin (1) and annonacin (2), is reported in full detail. Terminal acetylene 3 prepared from D-glucono delta-lactone and asymmetric dihydroxylation was employed as a common intermediate for both targets 1 and 2. Pd(0)-catalyzed coupling reaction of 3 with vinyl iodides 4 and 5, the chiral centers of which were taken from D-xylose and S-(-)-ethyl lactate, afforded enyne 26 and 27, respectively. Selective hydrogenation of 26 or 27 with diimide followed by removal of MOM ethers completed the synthesis of 1. A coupling reaction between the lithium derivative of 3 and epoxide 6 in the presence of boron trifluoride etherate gave 42. Both chiral centers in epoxide 6 were taken from L-ascorbic acid. Subsequent catalytic hydrogenation and MOM protection led to 43b. Introduction of the butenolide moiety by aldol condensation of protected S-lactal followed by cleavage of all MOM ethers completed the synthesis of 2. PMID- 11430105 TI - Unusual head-to-tail coupling of alkyl benzoates by electroreduction. AB - The electroreduction of alkyl benzoates in an alcoholic solvent gave unusual head to-tail coupled products. Usual head-to-head coupled products derived from acyloin condensation could not be detected. The best result (73% yield) was obtained from methyl benzoate using an undivided cell with an Sn cathode in i PrOH containing tetraalkylammonium salt as a supporting electrolyte. Using an undivided cell, the products cross-coupled with a solvent molecule were obtained as byproducts. The substitution at the para position of methyl benzoate considerably decreased the yields of the head-to-tail coupled products and increased those of the cross-coupled products. The possible mechanism of the head to-tail coupling is the attack of anion radical, generated from methyl benzoate by one-electron transfer, to another methyl benzoate. The cross-coupled products were formed by the reaction with carbonyl compound anodically produced from a solvent molecule. The cross-coupling between methyl benzoate and aromatic aldehydes was also effected by the mixed electroreduction under the same conditions. PMID- 11430106 TI - A polymer-supported proline-based diamine catalyst for the kinetic resolution of racemic secondary alcohols. AB - The preparation of polymer-supported proline-based diamine catalyst 12 for the kinetic resolution of racemic mixtures of secondary alcohols is described. Not only is the catalyst effective for the resolution of a host of different alcohols, it can also be recovered and reused several times without loss of either activity or selectivity. The catalyst has been used in conjunction with a polymer-supported sequestration strategy, giving rise to an essentially pure mixture of resolved products that can be separated using flash chromatography. PMID- 11430107 TI - A mild and general solid-phase method for the synthesis of chiral polyamines. Solution studies on the cleavage of borane-amine intermediates from the reduction of secondary amides. AB - A mild oxidative workup protocol using iodine in an acetic acid-acetate buffer solution is described for the cleavage of borane-amine adducts arising from the borane-promoted reduction of polyamides supported onto practical trityl-based resins. Chiral polyamines with diverse side-chain functionalities can be generated as free bases without premature release from the solid support and with essentially no racemization using this method. A series of model oligomeric secondary diamides 6 containing various alpha-amino acid residues (Val, Phe, Tyr, Ser, Cys, Met, Gln, Trp) provided triamine products 8 in high yields and good to excellent purity. On the other hand, a substrate containing a tertiary amide (15) formed a rather unusual triaminoborane intermediate that required more stringent workup conditions to liberate the polyamine product 20. The reduction of oligomeric tertiary amides such as 9 was found sluggish, but these compounds could nonetheless be obtained in high purity from in situ reductive amination of the corresponding secondary amines. Control studies, carried out in solution with model secondary amide 23, confirmed the efficiency of the buffered iodine solution and highlighted several advantages (no heating necessary, no need for strong bases or acids) over existing methods for the cleavage of borane-amine adducts. A possible mechanism involving all buffer components (iodine, acetic acid, and acetate ion) is proposed in which borane-amine adducts are transformed first to the monoiodoborane-amine and then to the corresponding acetoxyborane amine adduct of much weaker coordination affinity. The latter would dissociate readily and get trapped by the acetic acid to provide the desired secondary amine. This reduction/oxidative workup protocol is useful as a general method for the facile solid-phase synthesis of polyamines for eventual release in solution and use in various applications. It is also potentially very useful toward the synthesis and screening of bead-supported libraries of free oligoamines assembled through split-pool methods. PMID- 11430108 TI - Intramolecular reactions of benzylic azides with ketones: competition between Schmidt and Mannich pathways. AB - The Lewis acid-promoted reactions of benzylic azides with ketones can proceed by two major pathways. The azido-Schmidt reaction involves simple addition of azide to the ketone followed by rearrangement and ring expansion. In addition, benzylic azides can undergo prior rearrangement to afford iminium ions that can subsequently participate in a Mannich reaction. A series of ketones containing an alpha CH2(CH2)nCH(N3)Ph substituent (n = 1-3) was prepared to investigate the dependence of products on ketone ring size and tether length. For all ketones examined, good yields of bicyclic lactams arising from intramolecular Schmidt reaction were obtained when a four-carbon linker was used (n = 1 in the above formulation), but Mannich products predominated for the longer tethers examined (n = 2, 3). PMID- 11430109 TI - Diastereoselective tandem Michael-intramolecular Wittig reactions of a cyclic phosphonium ylide with 8-phenylmenthyl enoates. AB - The diastereoselective tandem Michael-intramolecular Wittig reactions of a five membered cyclic phosphonium ylide 2 using 8-phenylmenthyl enoates were examined. The reaction of the phosphonium ylide with 8-phenylmenthyl cinnamate followed by the hydrolysis of the resulting enol ether 4a afforded (3R,4S)-4 (diphenylphosphinyl)-3-phenylcycloheptanone (3R,4S)-5a as the major isomer. The diastereoselectivity of the initial tandem reactions was estimated to be 94:6 from the 31P NMR of a mixture of the diastereomeric ketal derivatives 6a and 6'a which were obtained by the reaction of 5a with (2R,3R)-2,3-butanediol, and the absolute configuration of the major isomer was determined by the single-crystal X ray analysis. Similar reactions using some 8-phenylmenthyl alkenoates were attempted. As a result, it was clarified that the corresponding trans-ketones 5b d were obtained and that the diastereomer ratios of their ketal derivatives were 60:40-73:27. PMID- 11430110 TI - Asymmetric aldol additions: use of titanium tetrachloride and (-)-sparteine for the soft enolization of N-acyl oxazolidinones, oxazolidinethiones, and thiazolidinethiones. AB - Asymmetric aldol additions using chlorotitanium enolates of N-acyloxazolidinone, oxazolidinethione, and thiazolidinethione propionates proceed with high diastereoselectivity for the Evans or non-Evans syn product depending on the nature and amount of the base used. With 1 equiv of titanium tetrachloride and 2 equiv of (-)-sparteine as the base or 1 equiv of (-)-sparteine and 1 equiv of N methyl-2-pyrrolidinone, selectivities of 97:3 to > 99:1 were obtained for the Evans syn aldol products using N-propionyl oxazolidinones, oxazolidinethiones, and thiazolidinethiones. The non-Evans syn aldol adducts are available with the oxazolidinethione and thiazolidinethiones by altering the Lewis acid/amine base ratios. The change in facial selectivity in the aldol additions is proposed to be a result of switching of mechanistic pathways between chelated and nonchelated transition states. The auxiliaries can be reductively removed or cleaved by nucleophilic acyl substitution. Iterative aldol sequences with high diastereoselectivity can also be accomplished. PMID- 11430111 TI - Stereodynamics of N-isopropyl-N-methylpropargylamine. Dynamic NMR studies. Molecular mechanics calculations. AB - N-Isopropyl-N-methylpropargylamine (N-isopropyl-N-methyl-2-propyn-1-amine; IMPA) is chiral at the pyramidal nitrogen. Racemization occurs via an inversion rotation process. Both 13C(1H) and 1H dynamic NMR (DNMR) spectra decoalesce in response to slowing inversion-rotation (delta G++ = 7.7 +/- 0.1 kcal/mol). While aspects of the DNMR spectra suggest the presence of minor conformations, the spectrum at 100 K shows a strong preference for one conformation. The NMR data suggest that the preferred conformation has both the isopropyl methine proton and the ethynyl group anti to the lone pair. Both isopropyl methyl groups are gauche to the lone pair. This conformational preference is in significant contrast to N ethyl-N-methyl-2-aminopropane in which the population of that conformation having the ethyl methyl group and the isopropyl methine proton both anti to the lone pair is only 5% at 95 K. The NMR data, supported by molecular mechanics (MMX) calculations, suggest a special stabilization for the ethynyl group being oriented anti to the lone pair. PMID- 11430112 TI - Opening of thiiranes: preparation of orthogonal protected 2-thioglyceraldehyde. AB - Treatment of acrolein diethyl acetal sulfide 8 with methanesulfenyl bromide at low temperature results in an efficient thiirane ring opening to a halo disulfide 9. The bromine in this halo disulfide is easily substituted by silver acetate, sodium azide, sodium iodide, and silver nitrate. Treatment of 9 with tetrabutylammonium acetate yields a novel dehydrohalogenation product 12. Silica gel converts bromide 9 into a disulfide-substituted version of acrolein 15. The orthogonal-protected version of 2-thioglyceraldehyde 13 can be deprotected to a useful form of this aldehyde. PMID- 11430113 TI - Quantification of the various contributors to rate enhancement in nucleophilic strain releasing reactions. AB - Since cyclopropanes are much more reactive than cyclobutanes despite their nearly identical strain energy, it has become clear that rate enhancement in cyclopropanes is partly due to strain release but also partly due to an additional factor about whose nature there is no consensus. Activation and equilibrium energies for a series of nucleophilic reactions of MeO-, MeS-, MeNH-, Me2N-, and MePH- with strained rings ranging from epoxide to aza-cubane were computed at the HF/631 + G* level. Using the Marcus equation, in combination with the computed intrinsic barrier for the identity reactions of the same nucleophiles with the strain free reference compounds (e.g., MeO- with Me-O-Me and Me2N- with Me3N), enabled a quantitative determination of the individual contributions of the partial strain release and the additional factor to the overall lowering of the transition state energy. Analysis of the data reveals the following: (a) There is no contribution of the additional factor to rate enhancement in four membered rings for first row elements (O and N) and only a small contribution (approximately 2 kcal) for second row elements (S and P). This is to be compared with a contribution of 7-17 kcal for three membered rings. (b) A significant synergistic effect is observed. Thus, in housane, for example, the additional factor amounts to 12.7 kcal which is more (by nearly 5 kcal) than the sum of the individual contributions of the isolated three and four membered rings. (c) The magnitude of the additional factor was found to be Periodic Table row dependent. PMID- 11430114 TI - Samarium(0) and 1,1'-dioctyl-4,4'-bipyridinium dibromide: a novel electron transfer system for the chemoselective reduction of aromatic nitro groups. AB - A mild and efficient electron-transfer method was developed for the chemoselective reduction of aromatic nitro groups using samarium(0) metal in the presence of a catalytic amount of 1,1'-dioctyl-4,4'-bipyridinium dibromide. This method was found to give the product aromatic amine in 79-99% yield with selectivity over a number of other functional and protecting groups such as alkene, azide, benzyl ether, nitrile, amide, halide, p-toluenesulfonamide, t-Boc, tert-butyldiphenylsilyl ether, and aliphatic nitro groups. Our results also indicate that samarium(0) plays an important role in the reduction process and that 1,1'-dioctyl-4,4'-bipyridinium dibromide acts as an electron-transfer catalyst and is essential in the activation of samarium(0) metal. The major active reducing agent responsible for the reduction is believed to be the radical cation species formed from 1,1'-dioctyl-4,4'-bipyridinium dibromide. PMID- 11430115 TI - Expedient access to the okadaic acid architecture: a novel synthesis of the C1 C27 domain. AB - A newly designed synthetic entry to the C1-C27 domain of okadaic acid has been developed. This incorporates substantial improvements in the preparations of the key okadaic acid building blocks representing the C3-C8, C9-C14, and C16-C27 portions. The synthesis of the C3-C8 lactone used (R)-glycidol as the origin of the C4 stereogenic center and featured a late-stage optional incorporation of the C7 hydroxyl group. The complementary C9-C14 fragment was synthesized in a concise route from (R)-3-tert-butyldimethylsilyloxy-2-methylpropanal and propargyl bromide. Assembly of the C3-C14 spiroketal-containing intermediate from the constitutent fragments revealed a dramatic effect of C7 functionalization upon spiroketalization efficiency. In contrast, both (9E)- and (9Z)-enones converged readily to the C8 spiroketal upon treatment with acid. Modifications to the central C16-C27 fragment of okadaic acid included the early replacement of benzylic protecting groups by more suitable functionalities to facilitate both the generation of the C15-C27 intermediate and the deprotection of the final products. These modular building blocks were deployed for the synthesis of the C1 C27 scaffold of 7-deoxyokadaic acid. This work demonstrates improvements in the formation of versatile okadaic acid intermediates, as well as a reordering of fragment couplings. This alternative order of coupling was designed to promote the late stage incorporation of nonnatural lipophilic extensions from the C27 terminus. PMID- 11430116 TI - Applications of phthalimide photochemistry to macrocyclic polyether, polythioether, and polyamide synthesis. AB - Irradiation of phthalimides which contain N-linked omega-trimethylsilylmethyl substituted polyether, polythioether, and polysulfonamide chains results in efficient production of the corresponding macrocyclic polyether, polythioether, and polysulfonamide products. These photocyclization reactions follow sequential single electron transfer (SET)-desilylation pathways. Only in the cases of phthalimides, bearing mixed ether-thioether N-substituents, do these excited state cyclization reactions proceed with lower degrees of regioselectivity. This is a result of competitive desilylation and alpha-to-sulfur deprotonation reactions of the zwitterionic diradical intermediates formed by initial SET. PMID- 11430117 TI - Synthesis of (-)-astrogorgiadiol. AB - Reaction of Rh2(S)-PTPA4 with the (R)-citronellol-derived alpha-diazo-beta ketoester 1 led to the formation of cyclic beta-ketoester 2 in 95% yield and 48% diastereomeric excess. The purity of 2 was increased to > 99% de after one crystallization. To demonstrate its utility in steroid total synthesis, the beta ketoester 2 was carried on to secosteroid (-)-astrogorgiadiol (3), a naturally occurring vitamin D analogue with antiproliferative properties. PMID- 11430118 TI - Synthesis of (+/-)-puraquinonic acid: an inducer of cell differentiation. AB - Puraquinonic acid (1) was synthesized from 2,5-dimethoxybenzoic acid by way of isochroman 2 and the indanone derivative 3, a Nazarov cyclization being used to construct the five-membered ring. PMID- 11430119 TI - Bis-2-oxo amide triacylglycerol analogues: a novel class of potent human gastric lipase inhibitors. AB - A novel class of potent human gastric lipase inhibitors, bis-2-oxo amide triacylglycerol analogues, was developed. These analogues of the natural substrate of lipases were prepared starting from 1,3-diaminopropan-2-ol. They were designed to contain the 2-oxo amide functionality in place of the scissile ester bond at the sn-1 and sn-3 position, while the ester bond at the sn-2 position was either maintained or replaced by an ether bond. The derivatives synthesized were tested for their ability to form stable monomolecular films at the air/water interface by recording their surface pressure/molecular area compression isotherms. The inhibition of human pancreatic and gastric lipases by the bis-2-oxo amides was studied using the monolayer technique with mixed films of 1,2-dicaprin containing variable proportions of each inhibitor. The nature of the functional group (ester or ether), as well as the chain length, at the sn-2 position influenced the potency of the inhibition. Among the compounds tested, 2 [(2-oxohexadecanoyl)amino]-1-[[(2-oxohexadecanoyl)-amino]methyl]ethyl decanoate was the most potent inhibitor, causing a 50% decrease in HPL and HGL activities at 0.076 and 0.020 surface molar fractions, respectively. PMID- 11430120 TI - Carbon-to-carbon identity proton transfers from propyne, acetimide, thioacetaldehyde, and nitrosomethane to their respective conjugate anions in the gas phase. An ab initio study. AB - Gas-phase acidities of CH3Y (Y: NO, C identical to CH, CH=NH, and CH=S), barriers to the identity proton-transfer CH3Y + CH2=Y- reversible CH2=Y- + CH3Y, as well as geometries and charge distributions of CH3Y, CH2=Y- and the transition states of the proton transfers were determined by ab initio methods at the MP2/6-311 + G(d,p)//MP2/6-311 + G(d,p), B3LYP/6-311 + G(d,p), and BPW-91/6-311 + G-(d,p) levels of theory. The acidities were also calculated at the CCSD(T)/6-311 + G(2df,2p) level. To make more meaningful comparisons, the same quantities for previously studied systems (Y: H, CH=CH2, CH=O, CN, NO2) were recalculated at the levels used in the present work. The geometric parameters as well as the group charges indicate that the transition states for all the reactions are imbalanced, although there is no correlation between the degree of imbalance and the pi acceptor strength of the Y group. Based on multi-parameter correlations with the field (sigma F), resonance (sigma R), and polarizability effect (sigma alpha) substituent constants, the contributions of each of these effects to the acidities and barriers were evaluated. For the Y groups whose sigma F, sigma R, and sigma alpha are unknown (CH=NH, CH=S, C identical to CH), a method for estimating these substituent constants is proposed. The barriers for the CH3Y/CH2=Y- systems are all lower than for the CH4/CH3- system; this contrasts with the situation in solution where the Y groups lead to an increase in the barrier. The reasons for this reversal are analyzed. We also make an attempt to clarify the issue as to why the transition states of these reactions are imbalanced, a question which continues to draw attention in the literature. PMID- 11430121 TI - Photo-Arbuzov rearrangements of 1-arylethyl phosphites: stereochemical studies and the question of radical-pair intermediates. AB - The direct UV irradiation of the 1-arylethyl phosphites 7, 8, and 9 was carried out in acetonitrile, benzene, and cyclohexane, as was the triphenylene-sensitized reaction of 9. Dimethyl 1-phenylethyl phosphite, 7, gives the photo-Arbuzov rearrangement product, dimethyl 1-phenylethylphosphonate (10), in 67% average yield and minor amounts (2%) of 2,3-diphenylbutane (11a) in quantum yields of 0.32 and 0.02, respectively. The photorearrangement of optically active, predominantly (R)-1-phenylethyl phosphite 7 (R/S = 97/3; 94% ee), at 35-40 degrees C proceeds with a high degree of stereospecificity at the stereogenic migratory carbon to give predominantly (R)-10 (R/S = 86/14, 72 +/- 2% ee). Use of the nitroxide radical trap TEMPO affords phosphonate 10, presumably all cage product, from predominantly (R)-7 (R/S = 97/3; 94% ee) in 64% yield (80% ee, R/S = 90/10). By contrast, the 1-(4-acetylphenyl)-ethyl phosphite, predominantly (S) 8 (S/R = 98/2, 96% ee), on direct irradiation gives the corresponding phosphonate (12) in only 20% yield along with dimer 11b in 40% accountability yield. Phosphonate 12 is nearly racemic (R/S = 52/48). Direct irradiation of predominantly (R)-9 (R/S = 98/2, 96% ee), a 1-(1-naphthyl)ethyl phosphite, results in a product distribution similar to that from predominantly (R)-7, but with a somewhat higher degree of retention of configuration in the product phosphonate 13 (R/S = 93/7, 86 +/- 3 ee). By contrast, the triplet triphenylene sensitized photorearrangement of largely (R)-9 (R/S = 98/2, 96% ee) leads to product distributions similar to those from direct irradiation of predominantly (S)-8 and is accompanied by almost total loss of stereochemistry in its product phosphonate, 13 (R/S = 51/49). The partial loss of stereochemistry on direct irradiation of 7 and 9 provides evidence for radical pair formation. Furthermore, these stereochemical results are diagnostic of the multiplicity of the initial radical pair formed. Values for kcomb/krot for the proximate free radical pairs from 7 and 9, derived experimentally, are severalfold larger than those for the proximate singlet pair from Ph2C=C=N-CHPhMe, corrected to 35 degrees C. The possibility that kcomb is increased for the pairs from 7 and 9 is proposed. PMID- 11430122 TI - Extending the scope of chromium-manganese redox-coupled reactions: a one-pot synthesis of benzoxazoles. AB - A critically important strategy for synthetic chemistry is the development of "domino" processes: those capable of concatenating multiple transformations into a single step. Such transformations not only provide an increase in synthetic efficiency, but also imply the development of a significant degree of mechanistic understanding. We report herein a new domino reaction, in which a chromium manganese redox couple is employed both to catalytically reduce an o-hydroxy nitroarene and to oxidatively cyclize a subsequently formed imine. We find that the reaction is most effective for starting o-hydroxy nitroarenes with a strongly electron-withdrawing group at the para position. PMID- 11430123 TI - Chemoenzymatic synthesis of pyrrolo[2,1-b]quinazolinones: lipase-catalyzed resolution of vasicinone. AB - A facile synthesis of bronchodilatory pyrrolo[2,1-b]quinazoline alkaloids by azidoreductive cyclization strategy employing TMSCl-NaI and bakers' yeast is described. Both the chemical and enzymatic methods are mild and take place at room temperature in good yields. Further, synthesis and resolution of vasicinone has been carried out by employing different lipases. It has been observed that lipase PS provides acetate of (S)-vasicinone in 98% ee. PMID- 11430140 TI - Accumulation of in-vivo fatigue microdamage and its relation to biomechanical properties in ageing human cortical bone. AB - Bone matrix accumulates microdamage in the form of microcracks as a result of everyday cyclic loading activities. In two very recent studies, which used conventional histological stains and light microscopy techniques, the amount of this in-vivo microdamage in the cortices of long bones has been shown to increase with age. These articles have suggested that in-vivo microcracks may have an effect on the material properties of the tissue. However, a precise quantitative relationship between the number of microcracks and the mechanical properties of these same bones has not been produced before, and in particular the way the microcracks may affect the stiffness, the strength or possibly the toughness of the tissue. This article presents an examination of the in-vivo microdamage in human bones by the use of laser scanning confocal microscopy, which offers better discrimination and allows examination of the cracks in-situ. Quantification of in vivo fatigue microcracks was performed by counting the microcrack numerical density and surface density in specimens for which we have previously derived a full set of mechanical properties as a function of age. It is shown that bone microdamage relates more to the toughness (measured by three different measures) of ageing bone tissue than to its stiffness and strength. The result allows us (i) to re-evaluate the fragility of ageing human bone and put more emphasis on its energy-related resistance to fracture than perhaps on its stiffness or strength and also (ii) to understand more fully the causal relationship and interactions between microcracks and tissue toughness. PMID- 11430141 TI - Dynamic experimentation on the confocal laser scanning microscope: application to soft-solid, composite food materials. AB - Confocal laser scanning microscopy (CLSM) is used to follow the dynamic structural evolution of several phase-separated mixed biopolymer gel composites. Two protein/polysaccharide mixed gel systems were examined: gelatin/maltodextrin and gelatin/agarose. These materials exhibit 'emulsion-like' structures, with included spherical particles of one phase (i.e. polymer A) within a continuous matrix of the second (i.e. polymer B). Compositional control of these materials allows the phase order to be inverted (i.e. polymer B included and polymer A continuous), giving four basic variants for the present composites. Tension and compression mechanical tests were conducted dynamically on the CLSM, with crack/microstructure interactions investigated using a notched compact tension geometry. Gelatin/maltodextrin composites exhibit a 'pseudo-yielding' stress/strain response in both tension and compression, when the gelatin-rich phase is continuous, which was attributed to debonding of the particle/matrix interface. This behaviour is significantly less apparent for both the gelatin/agarose composites, and the maltodextrin continuous gelatin/maltodextrin composites, with these materials responding in a nominally linear elastic manner. Values of the interfacial fracture energy for selected compositions of the two biopolymer systems were determined by 90 degrees peel testing, where a gelatin layer was peeled from either a maltodextrin or agarose substrate. For biopolymer layers 'cast' together, a value of 0.2 +/- 0.2 J m-2 was obtained for the fracture energy of a gelatin/maltodextrin interface, while a significantly higher value of 6.5 +/- 0.2 J m-2 was determined for a gelatin/agarose interface. The interfacial fracture energy of the two mixed systems was also determined following an indirect elastomer composite debonding model. An interfacial fracture energy of approximately 0.25 J m-2 was determined using this approach for the gelatin continuous gelatin/maltodextrin composite, which compares favourably with the value calculated directly by peel testing (i.e. approximately 0.2 J m-2). A somewhat higher value was estimated for the gelatin continuous gelatin/agarose system (1.0-2.0 J m-2), using this model, although there are severe limitations to this approach for this mixed gel system. In the present case, it is believed that the differing mechanical response of the two mixed biopolymer systems, when the gelatin phase is continuous, arises from the order of magnitude difference in interfacial fracture energy. It is postulated that polymer interdiffusion may occur across the interface for the gelatin/agarose system, to a significantly greater extent than for interfaces between gelatin and maltodextrin, resulting in a higher interfacial fracture energy. PMID- 11430147 TI - Competition for working memory among writing processes. AB - Narrative, descriptive, and persuasive texts were written by college students in longhand or on a word processor. Participants concurrently detected auditory probes cuing them to retrospect about whether they were planning ideas, translating ideas into sentences, or reviewing ideas or text at the moment the probes occurred. Narrative planning and longhand motor execution presumably were heavily practiced, freeing capacity for rapid probe detection. Spare capacity was distributed equally among all 3 processes, judging from probe reaction times, when planning demands were low in the narrative condition. When motor execution demands were low in the longhand condition, however, reviewing benefited more than planning. The results indicate that planning, translating, and reviewing processes in writing compete for a common, general-purpose resource of working memory. PMID- 11430148 TI - Induced false memories of having heard the unspoken: a demonstration. AB - In a mock public survey situation, many students who had been led to expect to be asked to name their academic majors or home towns later declared that they had been asked for that information even though they had not. Such illusory memories were even more common among subjects who had spontaneously volunteered the unasked-for information during the interview. Both results have important implications for reality-monitoring processes and errors. PMID- 11430149 TI - Memory of action events: the role of objects in memory of self- and other performed tasks. AB - Encoding action phrases by enactment produces better recall than hearing or reading the action phrase. This study examined whether enactment enhances memory relative to observing another perform the same action. Theories of the enactment effect suggest that the complexity of the action, here manipulated by varying the number of objects involved in an action, may determine whether enactment enhances memory relative to observation. The results revealed a consistent subject performed task advantage across all object conditions; the size of the effect did not vary with increasing task complexity. Additionally, items that included the use of an object were recalled better than those without objects. The results are consistent with the views of Engelkamp and Zimmer (1997) and Backman, Nilsson, & Kormi-Nouri (1993), who argued that the SPT effect is due to motor and/or sensory encoding. PMID- 11430150 TI - Strength and resistance to interference in practiced recognition: memory retrieval abilities investigated through latent structure modeling. AB - Participants (N = 811) practiced paired-associate recognition with and without an interference manipulation and then practiced a pattern recognition skill in which patterns discriminated had features in common. Structure models of the covariances in task reaction times indicated two factors or abilities. The first was a baseline factor, hypothesized to include the ability to strengthen traces and other abilities common to all tasks. The second was a resistance-to interference factor, or the ability to quickly retrieve associations with elements in common with nonretrieved associations. Further modeling on a subset of the sample (n = 434) showed the baseline factor to reflect a memory strength ability independent of other confounding abilities (e.g., motor, reading abilities). Both memory abilities are discussed broadly with respect to cognitive skill acquisition, controlled versus automatic processing, and activation. PMID- 11430151 TI - Controlled variables: psychology as the center fielder views it. AB - Perceptual control theory (PCT) views behavior as the control of perception. The central explanatory concept in PCT is the controlled variable, which is a perceived aspect of the environment that is brought to and maintained in states specified by the organism. According to PCT, understanding behavior is a matter of discovering the variables that organisms control. But the possible existence of controlled variables has been largely ignored in the behavioral sciences. One notable exception occurs in the study of how baseball outfielders catch fly balls. In these studies it is taken for granted that the fielder gets to the ball by controlling some visual aspect of the ball's movement. This article describes the concept of a controlled variable in the context of research on fly ball catching behavior and shows how this concept can contribute to our understanding of behavior in general. PMID- 11430152 TI - Establishing the experimenting society: the historical origin of social experimentation according to the randomized controlled design. AB - This article traces the historical origin of social experimentation. It highlights the central role of psychology in establishing the randomized controlled design and its quasi-experimental derivatives. The author investigates the differences in the 19th- and 20th-century meaning of the expression "social experiment." She rejects the image of neutrality of social experimentation, arguing that its 20th-century advocates promoted specific representations of cognitive competence and moral trustworthiness. More specifically, she demonstrates that the randomized controlled experiment and its quasi-experimental derivatives epitomize the values of efficiency and impersonality characteristic of the liberal variation of the 20th-century welfare state. PMID- 11430153 TI - The history of the Nederlandse Vereniging voor Hypnose (Netherlands Society of Hypnosis). AB - The foundation and history of the Nederlandse Vereniging voor Hypnose (Netherlands Society of Hypnosis or Nvvh) is described. The year 2001 marks the 70th anniversary of the Nvvh's creation. The article describes the accomplishments, leadership, and philosophy of the Society across the decades. Current professional and training directions are discussed. PMID- 11430154 TI - Differentiation of hypnosis and relaxation by analysis of narrow band theta and alpha frequencies. AB - Narrow band theta and alpha activity were recorded over anterior and posterior sites before, during, and after hypnosis in high and low hypnotically susceptible subjects (N = 16). In theta, high susceptibles had greater activity posthypnosis, otherwise there were no group differences. These findings common to low and high susceptibles suggest that theta is an index of relaxation that continues after hypnosis in highs. In alpha in high susceptibles, posterior power increased from the prehypnosis to hypnosis conditions and decreased posthypnosis. Exactly the converse effects were seen in lows. Furthermore, highs had greater alpha power than lows during both prehypnosis and hypnosis conditions, demonstrating an association of alpha with hypnotic susceptibility. The results indicate that, whereas theta indexes relaxation, alpha indexes the hypnotic experience and susceptibility. PMID- 11430155 TI - Posthypnotic responding: the relevance of suggestion and test congruence. AB - Thirty real, hypnotized subjects and 34 simulating, unhypnotized subjects were given either a suggestion to respond when they heard a cue (general) or a suggestion to respond when they heard a cue after hypnosis (posthypnotic). Half the subjects were given the cue during hypnosis (hypnotic test) and half were given it after hypnosis (posthypnotic test). Those who were given the cue during hypnosis were also given it after hypnosis. Between- and within-group comparisons were made of subjects' behavioral responses, latencies to respond, and ratings of experiential compulsion. The findings indicated that subjects' behavior and experience were influenced by congruence between information conveyed by the suggestion and the test about when and how they should respond. PMID- 11430156 TI - Hypnotically induced emotional numbing. AB - This study investigated the utility of a hypnotic suggestion to inhibit emotional response. High and low hypnotizable participants (N = 53) were administered a hypnotic induction, and half the participants were then administered a suggestion for emotional numbing. Participants were then presented with slides depicting neutral or disfigured faces. Participants who received the emotional numbing suggestion reported less responsivity to the disfigured faces than did those in the control condition, and this pattern was stronger for high than for low hypnotizable participants. Highs in the numbing condition displayed less overall distress in their facial expressions in response to the disfigured slides relative to those in the control condition. These findings suggest that hypnotic emotional numbing may be a useful paradigm in which to explore processes in emotion inhibition. PMID- 11430157 TI - Information processing during hypnotically suggested sex change. AB - During hypnotically suggested sex change, 36 real (12 virtuoso and 24 high hypnotizable) and 18 simulating (low hypnotizable) individuals listened to a story involving a male and a female character. They subsequently reported their experience and recall of the story. Virtuosos were less likely than highs and simulators to identify with the character consistent with their suggested sex. However, virtuosos recalled more information about the character consistent with their suggested sex than did highs and simulators. The authors discuss the findings in terms of attention and the selective processing of information during hypnosis. They conclude that character identification was not the major factor that influenced the recall of virtuosos and suggest that virtuosos may have processed aspects of the information in a more self-referential way and thus encoded and recalled it more effectively. PMID- 11430158 TI - Treating adolescent conversion disorders: are hypnotic techniques reusable? AB - When treating disabling conversion disorders in hospitalized adolescents, clinicians must act to restore function as rapidly as possible. After attempting to rule out physical causes for the symptoms and trying to find persuasive psychological reasons that the patient will accept and use to resolve the condition, the inpatient staff of a multidisciplinary therapeutic milieu must seek additional approaches to the care of these seriously ill individuals. This clinical forum reports the author's experience treating 2 young patients, a 12 year-old girl and a 15-year-old boy, with hypnosis. Therapists of every experience level find hypnotic techniques that work for them in a variety of patients, but are hypnotic techniques really reusable? The author reports what he learned once again. PMID- 11430159 TI - Freedom from smoking: integrating hypnotic methods and rapid smoking to facilitate smoking cessation. AB - Hypnotic intervention can be integrated with a Rapid Smoking treatment protocol for smoking cessation. Reported here is a demonstration of such an integrated approach, including a detailed description of treatment rationale and procedures for such a short-term intervention. Of 43 consecutive patients undergoing this treatment protocol, 39 reported remaining abstinent at follow-up (6 months to 3 years posttreatment). PMID- 11430160 TI - Integrating hypnosis into a comprehensive smoking cessation intervention: comments on past and present studies. PMID- 11430161 TI - Inspector general nominee has familiar name. Appointment of Rehnquist's daughter to HHS post raises some eyebrows. PMID- 11430162 TI - Back-pedaling. AMA rethinks unionizing efforts, cites recent ruling. PMID- 11430163 TI - Rising like a phoenix. Less may be more for reincarnated Paracelsus. PMID- 11430164 TI - A taxing matter. Baltimore council to vote on energy levy for not-for-profits. PMID- 11430165 TI - Lobbying magic. Hospital groups spend less on influence but manage to achieve more. PMID- 11430166 TI - Collateral damage. Providers face negative fallout from the war on drugmakers. PMID- 11430167 TI - Four found whistleblowing the best revenge. PMID- 11430168 TI - Therapy on target. Change in HCFA reimbursement helps advance a new radiation technology. PMID- 11430169 TI - Gaining strength. Calif. state bond insurance program hopes to attract healthier facilities. PMID- 11430170 TI - This shortage needs CPR. Studies at odds on whether there are enough nurses to go around. PMID- 11430171 TI - Glare of an unblinking spotlight. New HCFA chief tones down characteristic candor after media's baptism by fire. PMID- 11430172 TI - Vaccinations in women. AB - Obstetricians/gynecologists who deliver primary care have an opportunity to enhance the preventive care of their patients. At the first visit with each patient, the vaccination history should be taken. Systems should be developed to remind the physician and office staff of the need for influenza and pneumococcal vaccination, including prompts for patients. One should create or use forms already developed by the CDC to record vaccination data in the patient chart. Additional information is available on the Web at www.cdc.gov/nip provided by the National Immunization Program. The Facts and Comparisons section has a Web site with links to other sources: www.immunofacts.com. All current recommendations of the ACIP are included in the reference list. PMID- 11430173 TI - Diagnosis and management of headache. AB - Migraine and tension headaches are among the most common diagnoses in women's health. Secondary causes of headache such as brain tumor, subarachnoid hemorrhage, and meningitis are uncommon but must not be missed. A careful history and physical examination, use of diagnostic criteria, and certain facts about the serious causes of headache are the keys to diagnosis and treatment. Neuroimaging should be limited to patients displaying signs or symptoms of a secondary headache cause. Menstrual migraine can be managed similarly to nonmenstrual migraine. PMID- 11430174 TI - Fatigue in primary care. AB - Fatigue is a common problem in primary care that may represent a reaction to life problems or be a component of a disease state. A careful history, physical examination, and a few directed laboratory tests can usually allow the physician to differentiate between fatigue caused by depression, situational stress, or physical causes such as postviral or drug-induced fatigue, endocrine disorders, sleep disorders, infectious diseases, autoimmune disorders, or neurologic disease. Uncommonly, patients may have otherwise unexplained fatigue lasting 6 months or more that fulfills the criteria of chronic fatigue syndrome. A range of diagnostic skills coupled with a therapeutic physician-patient relationship will usually be successful in treating women with symptoms of fatigue. PMID- 11430175 TI - Depression and anxiety disorders. AB - Depressive and anxiety disorders are common problems facing obstetrician gynecologists. Although psychiatric disorders are equally common in men and women, women are at least twice as likely to present with depressive disorders and most anxiety disorders. The depressive disorders include major depression, dysthymia, seasonal affective disorder, and premenstrual dysphoric disorder. The anxiety disorders are panic disorder (with and without agoraphobia), generalized anxiety disorder, social phobia, obsessive compulsive disorder, and PTSD. One must diagnose and manage depressive and anxiety disorders during pregnancy, the purpureum, and while breastfeeding. General treatment principles include assessing suicide risk, psychotherapy, pharmacologic treatment, and an appropriate medical work-up for depressive and anxiety disorders. The SSRIs are the first-line treatment for most depressive and anxiety disorders because of data supporting their efficacy, the minimal need for dosage titration, the overall favorable side-effect profile, and the length of available clinical experience. Newer antidepressants, such as venlafaxine, bupropion, nefazodone, and mirtazapine, are options for patients unresponsive to, or intolerant of, the SSRIs. Treatment considerations include acute, maintenance, and continuation therapy, dosage regimens, adverse effects, and drug interactions. Specific guidelines are available for referring patients to a mental health specialist. PMID- 11430176 TI - Smoking cessation in pregnant women. AB - Cigarette smoking is considered the most significant modifiable cause of adverse pregnancy outcomes in US women. Smoking cessation in pregnant patients is one of the most effective ways to reduce negative pregnancy outcomes of fetal growth retardation, preterm delivery, and perinatal mortality. Research evidence documenting the effectiveness of health care provider interventions in smoking cessation has led to the PHS recommendation to screen and counsel every patient. Materials from the PHS and NCI are available to assist health care providers in developing a brief office-based intervention. Patient materials for distribution are also available. Increased use of office-based cessation strategies, of cessation programs in community-based interventions, and of pharmacologic therapies is necessary to remedy the public health burden of fetal exposure to tobacco. Relapse prevention methods using office-based and social support systems are important to improve long-term maintenance of cessation in women who quit smoking during pregnancy. PMID- 11430177 TI - Diagnosis and management of respiratory tract infections for the primary care physician. AB - Respiratory tract infections cause nearly half of deaths owing to infectious disease in the United States. This article has discussed the management of several common respiratory tract infections, with an emphasis on appropriate diagnosis and use of antimicrobial agents. Understanding the cause of various respiratory tract infections enables primary care physicians to avoid unnecessary antibiotic use, decreasing adverse effects owing to medications and preventing the rise in antimicrobial resistance. PMID- 11430178 TI - Asthma. AB - Asthma is an important and increasingly prevalent respiratory disease. Its proper diagnosis and treatment lie at the heart of improving asthma outcome. Unfortunately, asthma has many faces and is affected by many variables, many of them difficult to control. Like many chronic illnesses, asthma tends to affect the poor and less advantaged individuals in society. Heightened awareness among patients and physicians of the serious nature of the disease is needed to reduce the morbidity and mortality of asthma. Treatment clearly requires a multifaceted approach, including behavioral, environmental, social, and medical interventions, in which the National Asthma Education and Prevention Program guidelines provide a logical, step-wise, and effective approach. PMID- 11430179 TI - Hypertension. AB - This article reviews some of the salient points in the management of hypertension as recommended by the "Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure." New developments since publication of this 1997 report are also discussed. PMID- 11430180 TI - Dyspepsia in pregnancy. AB - Dyspepsia with or without nausea is common during pregnancy. Known ulcer disease, gastritis, and GERD may improve during pregnancy. Many women have a stoic and long-suffering posture during pregnancy owing to an unrealistic expectation concerning the teratogenicity of commonly used drugs. It is appropriate in medicine to alleviate pain and suffering when possible, and many drugs can be used safely and effectively to control upper gastrointestinal tract symptoms. When symptoms are persistent into the late second trimester, refractory to pharmacologic treatment, or severe, H. pylori infection, complications of ulcer disease, and underlying cancer should be suspected and sequentially ruled out. More timely treatment and work-up of nonobstetric disease during pregnancy is expected to lower perinatal complications. PMID- 11430181 TI - Lower gastrointestinal disease in women. AB - Gastrointestinal disease is common in women and therefore a commonly encountered entity in gynecologic practice. A new understanding of the underlying pathophysiology of the most common condition, irritable bowel syndrome, is changing both diagnosis and therapy. PMID- 11430182 TI - Anemia. AB - Anemia is an enormous problem worldwide that is worse in the developing countries but by no means lacking in developed countries. Women and children are disproportionately affected. Nutritional anemias, mainly caused by a lack of iron and folate, are largely correctable by dietary improvement, but this socioeconomic change may not be readily achieved. The inherited disorders, the hemoglobinopathies and thalassemias, in their heterozygous forms are beneficial to the affected populations. Prevention of the homozygous forms and the serious diseases that result can only be addressed by education. These conditions are seen in all types of medical practice ranging from neonatology to geriatrics and public health and are an ongoing concern to all physicians. PMID- 11430183 TI - Hypoglycemia. AB - Except in diabetic patients receiving insulin or sulfonylureas, hypoglycemia is a rare disorder. It is identified by modified Whipple's criteria consisting of neuroglycopenic symptoms, a blood glucose level equal to or less than 40 mg/dL, and relief of symptoms by glucose use. The sources of the body glucose are dietary intake, glycogenolysis, and [figure: see text] gluconeogenesis. The metabolism of glucose involves oxidation and storage as glycogen or fat. Causes of hypoglycemia include medications or toxins capable of decreasing blood glucose, disorders associated with fasting hypoglycemia, and postprandial hypoglycemic disorders. The most common type of hypoglycemia is insulin-induced hypoglycemia in diabetics. Insulinoma is rare; however, it is the most common hormone-secreting islet cell tumor. The diagnosis is made by the occurrence of hypoglycemia in the presence of symptoms of neuroglycopenia and inappropriately high levels of insulin and C-peptide. In hospitalized patients, the diagnosis is best made by prolonged fast. Most insulinomas are small and require invasive methods for precise localization. In surreptitious insulin use, hypoglycemia is associated with low plasma C-peptide. Postprandial hypoglycemia occurs in response to feeding and is generally caused by excessive insulin effect. It is seen in patients with postgastric surgery and rarely in early diabetes mellitus. Idiopathic postprandial hypoglycemia is rare and seems to be caused by subtle abnormalities of insulin response to food. Treatment of postprandial hypoglycemia consists of frequent small meals, with deletion of refined carbohydrate and increased protein intake. Primary treatment of insulinoma is surgical resection of the tumor. PMID- 11430184 TI - Management of type 2 diabetes. Evolving strategies for treatment. AB - Type 2 diabetes is the most prevalent form of diabetes, accounting for approximately 90% of cases. This article examines the current classification, diagnostic criteria for diabetes, and screening recommendations and provides a therapeutic strategy for improving glycemic control in patients with type 2 diabetes. PMID- 11430185 TI - Evaluation of the patient with a suspected thyroid disorder. AB - The principal types of thyroid disorders described herein can be recognized and evaluated clinically and biochemically by the obstetrician/gynecologist. Most patients with diffuse goiter should be referred for further evaluation and an RAI scan arranged while this consultation is awaited. Patients with solitary thyroid nodules should be referred for FNAB. Primary hypothyroidism without significant goiter can be treated without specialist referral, except in patients at high risk for ischemic coronary disease or arrhythmia. Primary hyperthyroidism, apart from subacute and postpartum thyroiditis, should be referred for specialist evaluation and definitive therapy. An RAI scan and uptake should be ordered, and beta-blocker therapy can be initiated as interim therapy in symptomatic patients. Patients with secondary (i.e., hypothalamic- or pituitary-based) thyroid dysfunction should always be referred for evaluation and treatment of the primary intracranial cause. There are several causes of abnormal thyroid function tests in apparently euthyroid patients, and the clinical context often provides important evidence of their nature. Clinical judgment is an important factor in knowing when to arrange specialist consultation. PMID- 11430186 TI - The effects of parallelism and prosody in the processing of gapping structures. AB - Two studies explored the processing of ambiguous sentences like Bill took chips to the party and Susan to the game, which may be assigned a gapping (Susan took chips) or a nongapping structure (Bill took Susan). The central question was what factors affect the ultimate interpretive preferences for these sentences. In a written questionnaire, sentences with greater parallelism between arguments in the positions of Bill and Susan received more gapping responses, though an overall bias toward the nongapping structure was seen. An auditory comprehension study showed that prosodic parallels between arguments also affected interpretation. In both experiments parallelism played a significant role in determining an interpretation, but the simpler structure, the nongapping structure, was preferred overall. PMID- 11430187 TI - Constrained emergence of universals and variation in syllable systems. AB - A computational model of emergent syllable systems is developed based on a set of functional constraints on syllable systems and the assumption that language structure emerges through cumulative change over time. The constraints were derived from general communicative factors as well as from the phonetic principles of perceptual distinctiveness and articulatory ease. Through evolutionary optimization, the model generated mock vocabularies optimized for the given constraints. Several simulations were run to understand how these constraints might define the emergence of universals and variation in complex sound systems. The predictions were that (1) CV syllables would be highly frequent in all vocabularies evolved under the constraints; (2) syllables with consonant clusters, consonant codas, and vowel onsets would occur much less frequently; (3) a relationship would exist between the number of syllable types in a vocabulary and the average word length in the vocabulary; (4) different syllable types would emerge according to, what we termed, an iterative principle of syllable structure and their frequency would be directly related to their complexity; and (5) categorical differences would emerge between vocabularies evolved under the same constraints. Simulation results confirmed these predictions and provided novel insights into why regularities and differences may occur across languages. Specifically, the model suggested that both language universals and variation are consistent with a set of functional constraints that are fixed relative to one another. Language universals reflect underlying constraints on the system and language variation represents the many different and equally-good solutions to the unique problem defined by these constraints. PMID- 11430188 TI - Variability in fricative production and spectra: implications for the hyper- and hypo- and quantal theories of speech production. AB - Fricative spectral data are compared with articulatory data from electropalatographic (EPG) recordings in an investigation of coarticulatory effects on the acoustic signal. Data were taken from CV tokens produced by four female speakers of Australian English. Results are presented for the coronal fricatives /theta s [symbol: see text] z [symbol: see text]/ in seven monophthong vowel contexts. The analysis consists of a comparison of spectral centre of gravity (COG) with EPG centre of gravity measured along the horizontal dimension. The correlation between the articulatory and the acoustic data is quite high. Overall, the sibilant fricatives show very little variability in production, while the nonsibilant dental shows a good deal of variability. This is reflected in the spectral output. It is also shown that the alveolar sibilants show more effect from vowel context than do the postalveolar sibilants. These results are interpreted as showing that coarticulatory resistance is indeed greater for sibilant fricatives, but that degree of tongue body raising inherent in the fricative's production must also be taken into account. The results for overall variability are discussed with reference to the Hyper- and Hypo- and Quantal Theories of speech production. It is suggested that sibilant fricatives do not lend themselves to the articulatory imprecision which, according to these theories, characterizes perceptually salient, and typologically common, speech sounds. PMID- 11430189 TI - Help or hindrance: how violation of different assimilation rules affects spoken language processing. AB - Four phoneme-detection studies tested the conclusion from recent research that spoken-language processing is inhibited by violation of obligatory assimilation processes in the listeners' native language. In Experiment 1, native listeners of German detected a target fricative in monosyllabic Dutch nonwords, half of which violated progressive German fricative place assimilation. In contrast to the earlier findings, listeners detected the fricative more quickly when assimilation was violated than when no violation occurred. This difference was not due to purely acoustic factors, since in Experiment 2 native Dutch listeners, presented with the same materials, showed no such effect. In Experiment 3, German listeners again detected the fricative more quickly when violation occurred in both monosyllabic and bisyllabic native nonwords, further ruling out explanations based on non-native input or on syllable structure. Finally Experiment 4 tested whether the direction in which the rule operates (progressive or regressive) controls the direction of the effect on phoneme detection responses. When regressive German place assimilation for nasals was violated, German listeners detected stops more slowly, exactly as had been observed in previous studies of regressive assimilation. It is argued that a combination of low expectations in progressive assimilation and novel popout causes facilitation of processing, whereas not fulfilling high expectations in regressive assimilation causes inhibition. PMID- 11430190 TI - Data reveal capitation's impact on physician compensation. PMID- 11430191 TI - Medicaid data help fill information void, strengthen risk contracting efforts. PMID- 11430192 TI - Hospital benchmarks show clinical quality, efficiency are hallmarks of cost effective care. PMID- 11430193 TI - Are inpatient cost increases linked to utilization or other factors? PMID- 11430194 TI - MedBytes. PMID- 11430195 TI - Court of last resort. TMA takes legal action to stop managed care abuses. PMID- 11430196 TI - The privacy debate. HIPAA regulations take effect but the battle is far from over. PMID- 11430197 TI - The state of public health. Experts fear the system is in peril. PMID- 11430198 TI - Local health authorities. Their work is valuable, but the system is uneven. PMID- 11430199 TI - The price of progress. Getting insurers to cover new procedures is often difficult. PMID- 11430200 TI - When plates collide. Texas medical student aids Indian earthwuake victims. PMID- 11430201 TI - Use of alternative and complementary therapies for pediatric asthma. AB - This survey of 48 multicultural parents of children with asthma identifies and compares alternative and complementary treatments used for asthma, and compares any potentially effective or harmful effects. Thirty-nine (81%) of the parents used at least one form of alternative or complementary therapy to treat their child's asthma. Nontraditional therapies included prayer, over-the-counter medications, herbal teas, vitamins, and massage. African-American parents were more likely to rely on prayer, and Hispanic parents were more likely to use herbal and massage therapies. Use of over-the-counter medications and vitamins was similar among groups. Three herbal remedies were potentially toxic: lobelia, possible pennyroyal mint, and tree tea oil. Medical histories for all patients should include inquiries into the use of alternative therapies. PMID- 11430203 TI - Spirituality is valuable. PMID- 11430204 TI - Border health and physicians. PMID- 11430205 TI - Dose intensity for breast cancer. AB - Despite nearly 20 years of study, the importance of chemotherapy dose intensity in breast cancer remains unclear. Substantial preclinical data suggest a dose response relationship, and consistent data document that recipients of substandard dosing have inferior outcomes. The use of increased dose-intensive therapies is costly, may require the use of hematopoietic growth factor support, and can result in significant increases in both short- and long-term toxicities. In patients with metastatic disease, increased dose intensity frequently results in increased response rates. However, these increased responses have not translated into consistent improvements in time to progression or overall survival benefit. In the adjuvant setting, increases in the dose intensities of alkylating agents and anthracyclines have failed to support the concept of dose escalation beyond standard doses. Certain subgroups of patients, such as those whose tumors overexpress HER2/neu, may derive a benefit from more dose-intensive therapies. Early results of randomized trials of high-dose chemotherapy in the treatment of metastatic breast cancer and adjuvant therapy for high-risk, early stage breast cancer, are provocative. However, the often conflicting data do not support the routine use of this modality outside of the study setting. PMID- 11430206 TI - Neoadjuvant strategies for pancreatic cancer. AB - Recent prospective and retrospective data suggest that the use of multimodality therapy combining pancreaticoduodenectomy with postoperative adjuvant chemotherapy (fluorouracil) and external-beam radiation therapy maximizes local tumor control and improves the length of survival in pancreatic cancer patients, compared with surgery alone. Since postoperative chemoradiation is often delayed in these patients due to the morbidity and prolonged recovery time associated with surgery, investigators are assessing the efficacy of administering chemoradiation before pancreaticoduodenectomy in patients with potentially resectable pancreatic adenocarcinoma. When given prior to surgery, chemoradiation is not delayed and patients found to have disease progression after chemoradiation are not subjected to an unnecessary laparotomy. PMID- 11430207 TI - Clinical trials referral resource. Epidermal growth factor receptor inhibitors in clinical trials. PMID- 11430208 TI - Recent developments in chemotherapy for bladder cancer. AB - Invasive bladder cancer is a chemotherapy-sensitive neoplasm. Historically, the development of cisplatin (Platinol)-based chemotherapy regimens has represented an important advance for patients with metastatic disease. More recently, investigations of new agents, such as gemcitabine (Gemzar) and paclitaxel (Taxol), have resulted in further options for these patients. Randomized trials comparing new regimens with cisplatin-based therapies have been initiated. The role of chemotherapy in the adjuvant and neoadjuvant settings is an area that is undergoing active investigation. The application of prognostic biological markers to risk-stratify patients has resulted in new avenues of investigation in these ongoing early disease trials. PMID- 11430209 TI - Surgical approach to organ preservation in the treatment of cancer of the larynx. AB - Nonsurgical approaches to preservation of the larynx in the treatment of laryngeal carcinoma include either radiation alone or chemotherapy and radiation in combination. In light of the common use of total laryngectomy, these nonsurgical approaches have often been referred to as "organ-preservation" strategies. Within the context of this old paradigm, clinicians viewed surgery as offering the patient local control, with the caveat that the price would either be the loss of the larynx or the creation of a disability in swallowing or speech. Over the last 10 years, there has been a renaissance in the surgical approaches to laryngeal preservation, as evident in the surgical literature. A new paradigm has emerged in which both the surgical and nonsurgical approaches have equal value in laryngeal organ preservation. Today, a broad spectrum of surgical organ-preservation techniques are available that result in excellent oncologic outcomes as well as excellent and consistent functional outcomes in terms of speech and swallowing. The aim of this article is to present the options for organ-preservation surgery for laryngeal cancer. PMID- 11430210 TI - Dicey drugs from abroad. The government gets serious about knockoffs. PMID- 11430211 TI - Volleys in the cancer war. Has the 30-year enterprise been wrongheaded from the start? PMID- 11430212 TI - Of algebra and anemia. A link between iron deficiency and math woes? PMID- 11430213 TI - Herpes tests give answers you might need to know. PMID- 11430214 TI - Nurses are overworked, stressed, and hard to find. PMID- 11430215 TI - Cancer patients get simple words to say how it hurts. PMID- 11430216 TI - A critical analysis of Health and Human Services' proposed health privacy regulations in light of the Health Insurance Privacy and Accountability Act of 1996. AB - Dr. Eddy inquires, through a detailed analysis of relevant statutory provisions, whether it is likely that the HIPAA legislation or the Standards for Privacy of Individually Identifiable Health Information Regulations (SPIIHI) proposed by HHS will efficiently and cost effectively defend the privacy issues they were intended to safeguard. His paper critically analyzes the extremely broad and expensive course of action and tactics HHS has chosen to employ in this effort to determine whether HHS' proposal is an efficient use of health care dollars. To introduce his analysis, Dr. Eddy addresses the general concept of medical privacy and attempts to define what aspects of privacy that HIPAA and SPIIHI should protect. He challenges whether the proposed regulations are focused on existing problems and whether HHS has exceeded legislative mandates. PMID- 11430217 TI - The attorney/client privilege: a fond memory of things past an analysis of the privilege following United States v. Anderson. AB - Mr. Mustokoff, Mr. Swichar, and Ms. Herzfeld address the rudiments of the attorney/client privilege, its crime-fraud exception, corporate compliance programs, the United States government's quest for voluntary disclosure, and how those principles have been affected by United States v. Anderson. PMID- 11430218 TI - Administrative channeling under the Medicare Act clarified: Illinois Council, section 405(h), and the application of congressional intent. AB - Mr. Cogan and Mr. Johnson discuss the judicial review provisions of the Medicare Act, codified at 42 U.S.C. section 405(g) and (h), and the Supreme Court cases including Illinois Council that interpret the scope of the "arising under" language. They also examine the history of section 205(h) of the Social Security Act, including the most recent amendment contained in the Deficit Reduction Act of 1984. The relationship between 205(h), DEFRA, and section 405(h) is explored as well as the caselaw addressing and applying section 405(h) in light of Congress' 1984 amendments. PMID- 11430219 TI - The anti-kickback statute standard(s) of intent: the case for a rule of reason analysis. AB - Mr. Aspinwall presents a comprehensive overview of the anti-kickback statute to show how Congress enacted the legislation to prevent inappropriate utilization and to reduce the ambiguity of the anti-kickback prohibitions by adding a knowing and willful requirement to the standard of intent. He discusses the different lines of caselaw on the anti-kickback standard of intent, and closely examines the standard endorsed by the OIG. He proposes adopting a standard of reasonableness modeled after the rule of reason from antitrust law, using cost effectiveness as the primary criterion. He argues that a cost-benefit outcomes based approach to the anti-kickback statute would better serve the public interest. PMID- 11430220 TI - Telemedicine: the invisible legal barriers to the health care of the future. AB - Telemedicine has the potential to transform the world of health care just as the Internet transformed the world of commerce. Ms. Daly examines two legal obstacles to expanding the use of telemedicine: licensure and liability. She defines telemedicine and discusses its common applications and significant benefits. Licensure laws and liability rules result in formidable barriers to the expanded use of telemedicine, while also failing to provide sufficient protection for consumers. Ms. Daly argues that for the benefits of telemedicine to reach those most in need, mutual recognition of licensing laws coupled with a universal standard of care is necessary. PMID- 11430221 TI - Perspectives. Can M+C work? Only if minds and expectations change. PMID- 11430222 TI - Virginia Mason reaps the benefits of a 'data-driven culture'. PMID- 11430223 TI - Sacramento hospital boosts outcomes by focusing on high-risk CHF patients. AB - Sacramento's Mercy Heart Institute turns to one-on-one counseling to reduce the risk of hospitalization among congestive heart failure patients. One innovative spin is the use of a hightech linkage to home called Health Hero. PMID- 11430224 TI - Cataract surgery data offer practice benchmarks. AB - The latest report from the research arm of the Accreditation Association for Ambulatory Health Care provides benchmarks for cataract surgery. PMID- 11430225 TI - Forward-thinking practices can overcome technophobia, enjoy benefits of new software. AB - The Medical Group Management Association set out to answer the question with a survey of almost 500 physician practices. It offers some explanations and an update on the state of computer use among physicians. PMID- 11430226 TI - Links between productivity, compensation growing. AB - Use these tools to design compensation packages that keep peace in the practice and a healthy bottom line. PMID- 11430228 TI - [A new surgical method: lengthening of jaw bone by means of distraction technique]. AB - Callus distraction, or distraction osteogenesis as the method also is called, is a new surgical technique which makes it possible to create new bone by bone elongation. The method was originally used in orthopedic surgery to extend long bones but has subsequently proved useful for extending the mandible as well as other bones in the facial skeleton. By callus distraction, it is now possible to treat severe jaw anomalies and also to treat bone defects in the jaws without bone transplantation. This article describes the biological background to the method and the basic principles that must be observed for successful results. The treatment of four patients illustrates the method. PMID- 11430227 TI - [Emergency administration of statins in hospital. Measures concerning life style of coronary patients are still not to be forgotten]. PMID- 11430229 TI - [Functional evaluation of heart failure patients in primary care. The NYHA system is unreliable since intercurrent illness can give rise to similar symptoms]. AB - A group of 158 patients with congestive heart failure was studied in detail concerning other types of morbidity. The purpose was to evaluate how often classification according to the NYHA system was disturbed by intercurrent disease with a similar set of symptoms. At the same time, a visual analogue scale for the evaluation of physical capacity was tested. Seventy-eight per cent of the patients had intercurrent disease putting them at risk of inadequate NYHA classification. There was no correlation between the patients' NYHA class and grade or type of echocardiographic ventricular dysfunction. However there was a correlation between NYHA class and the presence of intercurrent disease. PMID- 11430230 TI - [Case report. Ink intoxication--a man colored blue]. PMID- 11430231 TI - [Shortage of general surgeons in Stockholm within five years]. PMID- 11430232 TI - [Changed abortion policy in the USA will result in increased maternal mortality globally]. PMID- 11430233 TI - [Research ethics for researchers, students and patients on the Web]. PMID- 11430234 TI - [Children with mentally disabled parents--invisible but with extensive needs]. PMID- 11430235 TI - [Crisis of rehabilitation. Dismantling of resources]. PMID- 11430236 TI - [A reply on criminal policy: condemned report accepted by an influential journal]. PMID- 11430237 TI - ["Green islands" for primary health care in the Balkan?]. PMID- 11430238 TI - On seeing clearly. PMID- 11430239 TI - The Fourier theory of vision. AB - The historical roots of the Fourier theory of spatial visual perception are traced. The development of the underlying concepts and the psychophysical experiments that led to them, and that they in turn spawned, are examined, as well as their relation to the current knowledge of neural substrates in the retina and primary visual cortex. Allowing nonlinearities or even substituting other types of basis functions does not eliminate the difficulties faced by any theory of visual perception that is built on the notion of fixed spatial filters. PMID- 11430240 TI - Evaluating models of collinearity judgment for reliability and scale. AB - In prior work from this laboratory, we have examined how accurately subjects can judge collinearity, i.e. alignment, as a function of the angular position of stimulus elements. In those experiments, subjects were presented with a line segment (or a pair of dots) which varied across the 360 degrees range of angular positions, and were required to select and mark a point which was perceived as being collinear. We found that the models of error for each subject consisted of a complex set of peaks and valleys which are herein described as delta excursions. The error tendencies appeared to be idiosyncratic, in that each subject manifested a different profile of these excursions. Here we report the results of three experiments. In the first, we tested subjects across five sessions, and found that the five models of error were fairly comparable for a given subject. In the second and third experiments, we tested at close intervals of angular position, and found evidence for localized sources of error. We continue to find that the error model for each subject is idiosyncratic, and argue that the excursions may be due to defects in a system which combines responses from position-encoding fields that vary in size. PMID- 11430241 TI - On the interference of task-irrelevant hue variation on texture segmentation. AB - Although natural images often include discordant information about object boundaries, the majority of research on texture segmentation has involved variation along a single dimension, e.g. colour, orientation, size. In this study, we examined orientation-based texture segmentation in the presence and absence of task-irrelevant colour variation. Previously, it had been shown that orientation-based texture segmentation was impaired if the elements, normally of one colour, were randomly allocated one of two colours (Morgan et al, 1992 Proceedings of the Royal Society of London, Series B 248 291-295). We found that this interference disappeared, however, when the spatial pattern of the colour variation was regular, as opposed to random, and when the elements were randomly positioned. We consider four models of how relevant and irrelevant texture information might combine to produce the interference effect, with special regard to these new findings. None of the models could account for the dependency of the interference effect on the spatial arrangement of colour and orientation in the texture. We suggest that inter-element separation and spatial-frequency selectivity are critical variables in the interference effect. PMID- 11430242 TI - Visual discomfort: the influence of spatial frequency. AB - The response of different visual discomfort groups to a range of spatial frequencies at threshold and suprathreshold was investigated. In experiment 1, a paired-comparison task was conducted. The high visual discomfort group judged a spatial frequency of 4 cycles deg-1 as the most perceptually distorted and somatically unpleasant to view. The moderate and low visual discomfort groups judged 8 and 12 cycles deg-1 as more perceptually and somatically unpleasant to view than lower spatial frequencies. In experiment 2, the spatial contrast sensitivity function (CSF) for the high visual discomfort group was depressed for spatial frequencies between 1 and 12 cycles deg-1 in comparison with the moderate and low visual discomfort groups. When these same spatial frequencies were modulated at 6 Hz, CSFs were the same for all groups. These results are discussed in relation to a failure of inhibition across spatial-frequency channels in the high visual discomfort group. This may be explained by a more generalised parvocellular system processing deficit. Possible similarities between some forms of migraine and visual discomfort are highlighted. PMID- 11430243 TI - Visually induced reorientation illusions. AB - It is known that rotation of a furnished room around the roll axis of erect subjects produces an illusion of 360 degrees self-rotation in many subjects. Exposure of erect subjects to stationary tilted visual frames or rooms produces only up to 20 degrees of illusory tilt. But, in studies using static tilted rooms, subjects remained erect and the body axis was not aligned with the room. We have revealed a new class of disorientation illusions that occur in many subjects when placed in a 90 degrees or 180 degrees tilted room containing polarised objects (familiar objects with tops and bottoms). For example, supine subjects looking up at a wall of the room feel upright in an upright room and their arms feel weightless when held out from the body. We call this the levitation illusion. We measured the incidence of 90 degrees or 180 degrees reorientation illusions in erect, supine, recumbent, and inverted subjects in a room tilted 90 degrees or 180 degrees. We report that reorientation illusions depend on the displacement of the visual scene rather than of the body. However, illusions are most likely to occur when the visual and body axes are congruent. When the axes are congruent, illusions are least likely to occur when subjects are prone rather than supine, recumbent, or inverted. PMID- 11430244 TI - Haptic perception of the horizontal by blind and low-vision individuals. AB - We examined haptic perception of the horizontal in visually impaired people. Blind people (late blind and congenitally blind), persons with very low vision, and blindfolded sighted individuals felt raised-line drawings of jars at four angles. They had to demonstrate their understanding that water remains horizontal, despite jar tilt, by selecting the correct raised-line drawing given four choices. Low-vision subjects, with near perfect scores, performed significantly better than the other groups of subjects. While the late-blind and blindfolded sighted subjects performed slightly better than the congenitally blind participants, the difference between the late-blind and congenitally blind groups was nonsignificant. The performance of the congenitally blind subjects indicates that visual experience is not necessary for the development of an understanding that water level stays horizontal, given container tilt. PMID- 11430245 TI - Attractiveness of facial averageness and symmetry in non-western cultures: in search of biologically based standards of beauty. AB - Averageness and symmetry are attractive in Western faces and are good candidates for biologically based standards of beauty. A hallmark of such standards is that they are shared across cultures. We examined whether facial averageness and symmetry are attractive in non-Western cultures. Increasing the averageness of individual faces, by warping those faces towards an averaged composite of the same race and sex, increased the attractiveness of both Chinese (experiment 1) and Japanese (experiment 2) faces, for Chinese and Japanese participants, respectively. Decreasing averageness by moving the faces away from an average shape decreased attractiveness. We also manipulated the symmetry of Japanese faces by blending each original face with its mirror image to create perfectly symmetric versions. Japanese raters preferred the perfectly symmetric versions to the original faces (experiment 2). These findings show that preferences for facial averageness and symmetry are not restricted to Western cultures, consistent with the view that they are biologically based. Interestingly, it made little difference whether averageness was manipulated by using own-race or other race averaged composites and there was no preference for own-race averaged composites over other-race or mixed-race composites (experiment 1). We discuss the implications of these results for understanding what makes average faces attractive. We also discuss some limitations of our studies, and consider other lines of converging evidence that may help determine whether preferences for average and symmetric faces are biologically based. PMID- 11430246 TI - Rock's cognitive theory of illusory figures: a commentary. AB - Contrary to a previously published opinion, the 'cognitive' theory of illusory contours is rich in detail and empirical support. PMID- 11430247 TI - Last but not least. AB - New variations of the spiral illusion are demonstrated. They include spiral illusions of the Cafe Wall illusion and the Zollner illusion, as well as other new orientation illusions. Thus the spiral illusion is not limited to the Fraser illusion. We discuss the role that detectors of spirals in a higher visual area might play in the spiral illusion. PMID- 11430248 TI - Reexamining the evidence of an ecological association between income inequality and health. AB - Several recent studies have made the provocative claim that income inequality is an important determinant of population health. The primary evidence for this hypothesis is the repeated finding--across countries and across U.S. states--that there is an association between income inequality and aggregate health outcomes. However, most of these studies examine only a single cross section of data and employ few (or even no) control variables. We examine the relationship between income inequality and aggregate health outcomes across thirty countries over a four-decade span and across forty-eight U.S. states over five decades. In large part, our findings contradict previous claims. PMID- 11430249 TI - Relating social inequalities in health and income. PMID- 11430250 TI - When economists and epidemiologists disagree.... PMID- 11430251 TI - The impact of HMOs on hospital-based uncompensated care. AB - Managed care in general and HMOs in particular have become the vehicle of choice for controlling health care spending in the private sector. By several accounts, managed care has achieved its cost-containment objectives. At the same time, the percentage of Americans without health insurance coverage continues to rise. For profit and not-for-profit hospitals have traditionally financed care for the uninsured from profits derived from patients with insurance. Thus the relationship between growth in managed care and HMOs, hospital "profits," and care for the uninsured represent an important policy question. Using national data over an eight-year period, we find that a ten-percentage point increase in managed care penetration is associated with a two-percentage point reduction in hospital total profit margin and a 0.6 percentage point decrease in uncompensated care. PMID- 11430252 TI - HCFA and the states: politics and intergovernmental leverage. AB - In this article, we seek to explain variations in the Health Care Financing Administration's (HCFA) relationship to state governments. After reviewing several alternative models of the policy-making process, we argue that the utility of each model depends on certain issue characteristics, especially salience and conflict. We further argue that HCFA's choice of intergovernmental tools, rooted in a political setting, depends on the same issue characteristics. We illustrate our arguments by examining HCFA's behavior during the Clinton administration and by focusing on four cases: HMO performance measurement, nursing home regulation, lead screening for children, and the Children's Health Insurance Program (CHIP). PMID- 11430253 TI - Health insurance expansion through states in a pluralistic system. AB - The United States continues to stand almost alone among developed nations in its lack of universal health care coverage. In this essay, we argue that even though the debate over whether the federal government or states should lead the effort to expand health care coverage under the federal system is relevant in strategizing how to cover the uninsured; the more critical issues stem from the challenge of the mixed and fragmented mode of public-private financing of our pluralistic health care system. We base this argument on (1) an in-depth review of Oregon's and Tennessee's five years of experience with broad coverage reform in the context of the United States health care system and on (2) a more abbreviated review of other state experiences in providing health care coverage. We conclude from our review that when the will exists, states can substantially expand coverage. However, as one moves up the income scale, political support and resources are harder to come by. Further, concerns grow about the interface of public and private coverage, with issues of "crowd out" and other distributional questions dominating the discussion of coverage expansion as policy makers focus less on how to cover people than on how to make sure one kind of coverage doesn't preempt another. Concern for crowd out can then lead to policies that keep out some of the very people policy makers may want to cover. In this context the question whether states or the federal government is more likely to succeed in expanding coverage is eclipsed by the more fundamental challenges raised by pluralism. Neither federal nor state government is likely to be fully successful without first identifying ways of better coordinating public and private activities and resources to provide continuous and affordable coverage. PMID- 11430254 TI - Community-oriented primary care: a model for public health nursing. AB - The American Public Health Association defines public health nursing as the "practice of promoting and protecting the health of populations using knowledge from nursing, social, and public health sciences." In 1993, celebrating the centennial anniversary of its founding, nurse leaders recognized systemic changes have required nurses to function in clinical, illness-oriented roles rather than in their more traditional community and public health roles. With nurses' public health skills atrophying, these leaders urged members of the profession to eschew specialization and return to their generalist roots founded on the principles of community-based prevention and health promotion. Soon the Public Health Functions Project, designed in part to identify skills and curriculum needs of an array of practicing public health workers, examined the public health nursing profession. Its recommendations seek to ensure that public health nurses are trained to respond to current challenges that face public health. In this essay, we describe how a fellowship program that predated this national project by almost a decade anticipated the recommendations for shaping public health nursing by enrolling midcareer nurses in a program that taught the principles and practice of community-oriented primary care. Such principles represent a merger of clinical care with population health sciences; its more recent expressions teach clinicians to work as partners with communities to identify and address health problems. In reporting on this program, we show how nurses in practice can embrace their generalist roots, meet current challenges, and play a lead role in realizing the nation's goals for the year 2010. These aims incorporate recent recommendations for preparing public health nurses for change in the health care system. PMID- 11430255 TI - Understanding mutual benefit societies, 1860-1960. Essay review. PMID- 11430256 TI - Surgery and the patient with arthritis. AB - Patients with rheumatologic disease (e.g., arthritis) require special attention when surgery is considered. Proactive attention to medication usage, corticosteroid coverage, airway, respiratory, joint stability and fragility challenges, cold exposure, and coagulation risks facilitate surgical intervention for such individuals. PMID- 11430257 TI - Oxidative stress and mitochondrial damage in coronary artery bypass graft surgery: effects of antioxidant treatments. AB - We examined antioxidant actions in 73 patients undergoing coronary artery surgery by assessing mitochondrial damage and oxidative stress in ventricular biopsies obtained at preischemia and postreperfusion. Those patients who received antioxidant therapy benefited by less oxidative stress and mitochondrial damage. PMID- 11430258 TI - Diogenes syndrome: the self-neglect elderly. AB - Diogenes syndrome (DS) is characterized by severe self-neglect, domestic squalor, social withdrawal, syllogomania, and refusal of help that may be precipitated by stressful events. Secondary DS is related to mental disorders. Its management includes day care center and community care. PMID- 11430259 TI - Mechanism of human accommodation as analyzed by nonlinear finite element analysis. AB - Results of nonlinear finite element analysis support the Schachar theory of accommodation and demonstrate that the long-held Helmholtz theory of accommodation is impossible. PMID- 11430260 TI - The association between asthma, asthma, therapeutic classes and hepatic enzyme elevation among adult HMO members. AB - The results of this case-control study conducted at a large HMO support the notion that severe asthma may be associated with transient hepatic enzyme elevation. Conversely, we found no evidence of associations between asthma therapeutic classes and hepatic enzyme elevation. PMID- 11430261 TI - Acute myocardial infarction 2000 recognition. AB - Acute myocardial infarction occurs in two forms: unheralded attacks and those preceded by unstable angina. As the leading cause of death in the US, accounting for over 95 billion dollar annual cost, acute myocardial infarction requires up to-date recognition and management. PMID- 11430262 TI - Ocular sebaceous carcinoma: case report and review of the literature. AB - We report a case of ocular sebaceous carcinoma, involving incidental discovery in an elderly woman and its recurrence following inadequate eradication. We review the clinical presentation and treatment options with emphasis on potential errors in clinical judgment leading to recurrence. PMID- 11430263 TI - Use of oral montelukast in the treatment of asthma. AB - Montelukast, a new leukotriene modifier, has several benefits in the treatment of asthma in adults and children including improved relief of asthma symptoms, rapid onset, a safety profile comparable with placebo, and oral, once-daily dosing means excellent adherence. PMID- 11430264 TI - Recent advances in pulmonary embolism diagnosis and management. AB - Pulmonary embolism remains a major problem in clinical practice. Pulmonary angiography remains the most accurate diagnostic procedure. Standard therapy includes heparin then warfarin, and now low molecular weight heparin. The role of prophylactic measures cannot be over emphasized. PMID- 11430265 TI - The female menopause--now and in the millennium: new treatment options for a better quality of life. AB - Estrogen has traditionally been the only agent effective for the prevention and treatment of post-menopausal osteoporosis. Estrogen's positive effects are often outweighed by negative ones. Newer agents provide safer alternatives that should increase compliance and improve quality of life. PMID- 11430266 TI - Proceedings of the International Symposium on Colorectal Cancer. St. Gallen, Switzerland, December 4, 1998. PMID- 11430267 TI - Proceedings and abstracts of the 3rd Congress of the Asian Society for Stereotactic, Functional and Computer-Assisted Neurosurgery. Seoul, Korea, June 13-16, 1999. PMID- 11430268 TI - A review of the causes of central pontine myelinosis: yet another apoptotic illness? AB - One of the well recognized stimuli for central pontine myelinosis (CPM) is the rapid correction of chronic hyponatraemia. Conventionally this has been perceived to lead to pontine glial cell swelling through osmosis and eventually to cell death. However, although a purely osmotic argument has been central to any patho physiological understanding of CPM, there are deficiencies in this approach that do not account for why certain individuals develop CPM with relatively mild osmotic insults. Here we review the varying aetiologies of CPM and propose a novel hypothesis for CPM causation by suggesting that individuals predisposed to CPM have inadequate energy provision as well as other factors that result in a pro-apoptotic drive, which renders them susceptible to brain injury from diverse causes. In CPM, the precipitant of brain injury appears to be osmotic stress. Furthermore, this model suggests a number of therapeutic interventions that may prevent or at least mitigate the consequences of CPM. PMID- 11430269 TI - A brief neuropsychological assessment for the differential diagnosis between frontotemporal dementia and Alzheimer's disease. AB - The neuropsychological performance (including measures of language, semantic memory, visual and spatial perception and executive functions) of a group of 14 patients with the clinical diagnosis of probable frontotemporal dementia was compared with that of a group of 14 patients with a clinical diagnosis of probable Alzheimer's disease. The aim was to identify a specific cognitive profile of frontotemporal dementia, which could be used to select a sensitive, short evaluation for the differential diagnosis with Alzheimer's disease. Both groups were severely impaired in most tasks, including those 'frontal lobe' tests which have been suggested to play an important role in differential diagnosis. Significant differences were found only for a minority of tests (oral praxis, visual-spatial perception, and verbal fluency). A logistic regression showed that a shortened testing procedure based on four tests (Rey-Osterreith complex figure test recall, phonemic fluency, oral apraxia, and cube analysis) achieved a 70% sensitivity and 80% specificity for the correct classification of patients in the frontotemporal dementia or Alzheimer's disease group. In conclusion, a brief neuropsychological evaluation including these four tests, as well as other measures sensitive to the frontal impairment, can be useful in the differential diagnosis between the two pathologies, along with the clinical data. PMID- 11430270 TI - Ischemic stroke of unusual cause: clinical features, etiology and outcome. AB - The clinical features, etiology and neurological outcome of ischemic stroke of unusual cause (ISUC) have rarely been reported. We retrospective reviewed all patients with this stroke subtype entered in the Sagrat Cor Hospital of Barcelona Stroke Registry, which includes data from 2000 consecutive first-ever stroke patients admitted to the hospital between 1986 and 1995. Patients with previous ischemia and/or hemorrhagic stroke were excluded. Topographic, anamnestic, clinical and neuroimaging characteristics of ISUC were assessed. Predictors of this stroke subtype were determined by logistic regression analysis. Ischemic stroke of unusual etiology was diagnosed in 70 patients (32 men and 38 women), with a mean +/- SD age of 52 +/- 22.4 years. This stroke subtype accounted for 4.3% of all first-ever strokes and 6% of all first-ever brain infarcts. Etiologies included hematological disorders in 17 cases, infection in 11, migraine stroke in 10, cerebral infarction secondary to venous thrombosis in nine, primary inflammatory vascular conditions in six and miscellaneous causes in 17. In the multivariate analysis after excluding cerebral venous thrombosis (n = 9) and arterial dissection (n = 4), because of typical clinical and radiological features, independent predictors of ISUC included 45 years of age or less (odds ratio [OR] 14.8), seizures (OR 6.8), headache (OR 5.2), hemianopia (OR 2.6) and occipital lobe involvement (OR 3.0). Patients with ISUC presented a lower in hospital mortality rate (7.1% vs. 14.4%; P < 0.05), were more frequently symptom free at discharge (35.7% vs. 25.80%; P < 0.05) and experienced a longer mean length of hospital stay (23.7 days vs. 18.2 days; P = 0.06) than non-ISUC patients. We conclude that ISUC is infrequent, etiologies are numerous and hematologic disorders are the most frequent cause. We emphasize the better prognosis and the need to distinguish it from other ischemic stroke subtypes which have a different treatment approach and outcome. PMID- 11430271 TI - Thromboembolism: new insights and future prospects. Proceedings of the 7th International Symposium on Thromboembolic Disease. Palma de Mallorca, June 9-10, 2000. PMID- 11430272 TI - Proceedings of the 6th International Symposium on Clinical Evaluation of Drug Efficacy in Urinary Tract Infection and the Current Issues in the Management of UTI Symposium of the Commission for Treatment of Urinary Tract Infection. 21st International Congress of Chemotherapy. 4-7 July 1999, Birmingham, United Kingdom. PMID- 11430273 TI - Abstracts of papers presented at the Japanese-European Kobe Symposium on Thrombosis. 6-7 November 2000, Kobe, Japan. PMID- 11430274 TI - Proceedings of the 2nd Forensic Y Chromosome User Workshop. Berlin, Germany, June 16-17, 2000. PMID- 11430275 TI - Society for Academic Emergency Medicine 2001 annual meeting abstracts. PMID- 11430276 TI - Living and coping with excessive infantile crying. AB - AIM: The aim of the study was to elicit from parents how they lived and coped with a baby who cries excessively. It sought to identify factors which made coping more difficult, as well as interventions perceived by parents to be effective in improving the situation. BACKGROUND: The excessive crying of a baby can be one of the most trying aspects of parenthood. Although there have been many studies into the cause and treatment of the problem, these have produced little convincing evidence and even less agreement among investigators. In the light of this an alternative approach is to provide effective support to enhance coping until the crying abates. DESIGN: This study was based in pragmatic ethnography and followed an adapted grounded theory approach. A total of 25 adults from 14 families participated, recounting their experiences with 20 babies who either were currently crying excessively or had done so previously while less than 1 year old. Data was collected by a factual questionnaire and tape-recorded focused interviews conducted in the family home augmented by periods of participant observation. OUTCOMES: Almost every aspect of family life was disrupted, resulting in strained relationships, feelings of guilt, and concerns about losing control. Repeated attempts had to be made to establish a diagnosis and to find a cure. Such attempts failed, leading to a repeated cycle of hope and disappointment. The greatest potential to effect a positive impact lay in the supportive role of health visitors. They needed to visit frequently; stay for a prolonged period; demonstrate engagement with the family and its difficulties; and impart specific messages with conviction and sincerity. CONCLUSION: A specific, vital role was found for the health visitor in supporting parents through this problem, and aspects of the role were identified in which further study is indicated. PMID- 11430277 TI - Effectiveness of breast self-examination nursing interventions for Taiwanese community target groups. AB - BACKGROUND: Because of the increasing morbidity and mortality from breast cancer, the Health Department in Taiwan in 1992 has identified early detection of breast cancer as one of the national health objectives. The effectiveness of Breast Self Examination (BSE) in detecting breast cancer depends not only on frequency, but also on accuracy. This study explored the effects of nursing intervention on the BSE intention, BSE frequency and BSE accuracy among beauticians as community target groups. DESIGN: This study was comprised of two stages. In the first stage, 198 beauticians in Taipei County were surveyed. Those women with perceived barrier scores > or =15 points in the first stage were selected for the second stage with the quasi-experimental design. The experimental group was provided with BSE instruction for the enhancement of BSE competence, personalized strategies to reduce barriers to BSE and monthly telephone reminders after BSE instruction class for 3 months. The study instruments were Champion's BSE questionnaire, Lashley's 15 BSE steps and BSE items of social norm referenced Lierman et al. RESULTS: The mean age of the 198 beauticians was 28 (+/-8.6), the mean year of education was 10 (+/-2.4) and 60% of participants were married. The results of the study indicate that the program significantly increased BSE frequency, BSE accuracy, perceived benefit of BSE, perceived competence in BSE and decreased perceived susceptibility to breast cancer and perceived barriers to practice BSE. Multiple regression analysis indicated that the perceived competency, susceptibility and social normative influence accounted for 25% of the variance in BSE intention. Perceived competency and social normative influence accounted for 15% of the variance in BSE frequency, while intention, social normative influence and perceived competency accounted for 21% of the variance in BSE accuracy. PMID- 11430278 TI - The feasibility of a nurse practitioner-led primary health care clinic in a school setting: a community needs analysis. AB - AIM: The aim of this New Zealand study was to determine the feasibility of establishing a nurse practitioner-led, family focused, primary health care clinic within a primary school environment as a means of addressing the health needs of children and families. A secondary aim was to ascertain whether public health nurses (PHNs) were the most appropriate nurses to lead such a clinic. METHOD: Utilizing a community needs analysis method, data were collected from three sources --known demographic data, 17 key informant interviews and two focus group interviews. Questions were asked regarding the health needs of the community, the perceptions of participants regarding the role of the PHN, and the practicalities of establishing a clinic including the services participants would expect a clinic to provide. Analysis was exploratory and descriptive. RESULTS: Findings included the identification of a wide range of health issues. These included asthma management and control issues, the need to address poor parenting, and specific problems of the refugee and migrant population. Findings also demonstrated that participant understanding of the role of the PHN was less than anticipated and that community expectations were such that for a PHN to lead a primary health care clinic it would be likely that further skills would be required. Outcomes from investigating the practicalities of establishing a nurse practitioner-led clinic resulted in the preparation of a community-developed plan that would serve to address the health needs of children and families in the area the study was undertaken. Services that participants identified as being appropriate included health information, health education, health assessment and referral. CONCLUSION: Overall findings indicated that the establishment of a nurse practitioner-led, family focused, primary health care clinic in a primary school environment was feasible. While a PHN may fulfil the role of the nurse practitioner, it was established that preparation to an advanced level of practice would be required. PMID- 11430279 TI - What role do nurses play in Type 2 diabetes care in the community: a Delphi study. AB - OBJECTIVES: To identify the views of nurses on their current and future roles in care in the community for people with Type 2 diabetes. METHOD: A two-round Delphi study was used to elicit, and assess consensus on opinions from random samples of practice nurses, with a substantial role in the management of diabetes patients and diabetes specialist nurses. Both groups had defined levels of expertise in diabetes care and worked in Great Britain. The study instrument in round one comprised a questionnaire with 10 statement headings inviting opinions on the nursing role in the management and care of people with Type 2 diabetes. In the second round levels of agreement were elicited from responders on collated opinions from round one. The initial questionnaire also included closed questions to obtain a profile of current care delivered. RESULTS: Responders were 97 practice nurses and 69 diabetes specialist nurses with second round response rates of 93% and 86%, respectively. The 47 themes raised could be grouped under headings of clinical practice, responsibility, communication, knowledge, education/experience, rewards, and patients' roles. Many were common to several statements and both nurse groups. The level of agreement on the themes fed back was high, 85% of the 165 items achieved greater than 80% agreement. Agreement occurred for factors relating to clinical effectiveness, audit, knowledge, education and experience. Dissent occurred both within and between individual nurse groups on issues of responsibility, prescribing, service delivery, resources, and patient factors. CONCLUSIONS: This Delphi study has identified key areas of consensus on the management of care for people with Type 2 diabetes in the community but also areas in which further work needs to be carried out to understand the differences found. Many of the discordant items relate to the separate roles and associated issues of the two nurse groups. PMID- 11430280 TI - Levels of compliance shown by hypertensive patients and their attitude toward their illness. AB - AIM: The aim of this paper is to describe hypertensive patients' compliance by finding out more specifically what compliance, intention, activity, responsibility and collaboration mean in the treatment of hypertensive patients. A further aim was to clarify their attitudes toward their illness. RATIONALE: This study is part of a project titled 'Patients' self-motivated control of hypertension. METHOD: The intervention consists of a multimedia computer program and effective lifestyle counselling. Nine male and 12 female patients from one municipality were included in an intervention programme. Data were collected through thematic interviews and analysed using content analysis. FINDINGS: Four levels of compliance were found: subconscious, cognitive, action and attitudinal level. Patients also had four different ways of feeling about hypertension and its treatment: some were careless, some were serious, some adjusted well and yet others felt frustrated. CONCLUSION: Health care professionals should recognize each patient's way of complying with his/her treatment and his/her attitude toward the illness. Different patients need different kinds of care and support. In addition health care professionals should also recognize their own attitudes toward hypertension and the patient's position in the health care system. PMID- 11430281 TI - The haemodialysis machine as a lifeline: experiences of suffering from end-stage renal disease. AB - AIM: The aim of this study was to describe patients' experiences of suffering from end-stage renal disease (ESRD). RATIONALE: The rationale was to investigate how persons find meaning or make sense of their situation and how they experience suffering. The theoretical basis for the study was to view suffering at three levels. The first level was related to sickness and treatment. The second level was related to the care provided and the third level was related to each person's unique life experience and existence. METHOD: Data were collected by interviews focusing on questions concerning daily life, needs, and expectations for the future. A qualitative interpretative content analysis was used. Fifteen patients between the ages of 50-86 participated in the study. FINDINGS: Two main themes were identified describing these patients suffering. The first theme, 'the haemodialysis machine as a lifeline' consisted of three subthemes: 'loss of freedom', 'dependence on the caregiver', and 'disrupted marital, family and social life'. The second theme 'alleviation of suffering' consisted of two subthemes: 'gaining a sense of existential optimism' and 'achieving a sense of personal autonomy'. CONCLUSION: This study indicated that, in the lives of patients on haemodialysis, the main areas of suffering were related to loss of freedom expressed as dependence on the haemodialysis machine as a lifeline and, the caregivers. This time-consuming and tiring dependence affected marital, family and social life. Alleviation of suffering could be achieved by accepting dependence on the haemodialysis machine and maintaining autonomy by being seen as an individual by the caregivers. PMID- 11430282 TI - Fragmented sleep and tiredness in males and females one year after percutaneous transluminal coronary angioplasty (PTCA). AB - AIM: Reported sleep quality was examined in relation to anxiety, depression, selected psycho-physiological sleep disturbing symptoms, daytime dysfunctions according to poor sleep, and quality of life 1 year after percutaneous transluminal coronary angioplasty (PTCA). Furthermore, females were compared with age-matched males to examine any differences in sleep quality, psycho physiological symptoms and quality of life. STUDY DESIGN: A descriptive comparative study. SAMPLE: Twenty-two females, mean age 62.7 (SD 5.7) and 70 men, mean age 60.4 (SD 6.7), were consecutively recruited. METHOD: A mailed questionnaire was distributed, including the Uppsala Sleep Inventory, Spielberger State Anxiety scale, Zung's Self-rating Depression Scale and the Nottingham Health Profile instrument. RESULTS: Forty-six per cent of the patients had a history of myocardial infarction and 40.2% hypertension. Seventy-two per cent of the patients were anxious and 30.4% depressed. Difficulty in getting to sleep was related to intrusive thoughts with increased feelings of anxiety and sensitivity. Difficulty in maintaining sleep was the most common sleep complaint (42.4%) and physical tiredness/fatigue was the greatest consequence of disturbed sleep (51.1%). Those with pain, palpitations or respiratory nocturnal sleep-disturbing symptoms were characterised by shorter sleep duration, emotional distress, cognitive dysfunction, tiredness and reduced quality of life. Greater difficulties initiating sleep and worse health related quality of life were revealed in females compared with aged-matched males. CONCLUSION: Fragmented sleep is a problem partly because of psycho-physiological symptoms 1 year after PTCA, with reduced resilience to stress, increasing vulnerability or diminished coping ability and poorer quality of life. PMID- 11430283 TI - Endogenous and exogenous research? Findings from a bibliometric study of UK nursing research. AB - AIMS OF THE STUDY: This paper uses the findings of a recent bibliometric analysis of published UK nursing research to ask whether the field is characterized by a fundamental split between two underlying areas of research interest. These can be termed 'endogenous' and 'exogenous'. The former term describes research which tends to be concerned with problems and issues to do with nursing as a profession; the latter is concerned with problems and issues centring around the nursing of patients. DESIGN/METHODS: Papers in the Wellcome Trust's Research Outputs Database (ROD), a database of UK biomedical research, were analysed. Nursing papers published between 1988 and 1995 numbered 1,845, just less than 1% of the total papers in the ROD. RESULTS/FINDINGS: Analysis of the subfield identified that nursing research was atypical of biomedical research as a whole in a number of ways. One difference was that usually in biomedical research there is a general correlation between numbers of funders acknowledged on a paper, numbers of authors, and esteem of the journal in which a paper appears. In nursing there was, if anything, a tendency for highly esteemed papers to have fewer authors and be less likely to have acknowledged funding. However, the apparently endogenous and exogenous papers have quite different characteristics. This paper explores this apparent difference and possible reasons for this difference and will briefly compare nursing research with some other newly emerging social and academic groups. CONCLUSIONS: Thinking of nursing research outputs in this way can provide insight into the existence of different reward systems influencing nurse researchers. However, it is impossible to draw too confident a differentiation without reading each individual paper and making judgements about whether they are 'endogenous' or 'exogenous', a practice generally beyond the scope of bibliometric practice. PMID- 11430284 TI - Risk factors for citation errors in peer-reviewed nursing journals. AB - BACKGROUND: Citation errors are common among nursing journals. But, there are no data regarding the factors that predispose to these errors. OBJECTIVE: To determine the risk factors that predispose to citation error in peer-reviewed nursing journals. METHODS: Five hundred and fifty references were selected randomly from articles published in eleven nursing journals for the year 1998. The incidences of major and minor citation errors were determined by comparing with the original articles. The relative odds of citation errors for the number of authors, collaborating institutions and the length of the reference list were calculated. The correlation between the scientific quality of the journal (by means of journal impact factor and immediacy index) and the incidence of citation error were also determined. RESULTS: The incidence of citation errors is comparable to those reported previously. Long reference lists in articles written by a single author predicted strongly the occurrence of minor citation errors. Journals with a high impact factor and immediacy index tend to contain fewer minor mistakes. None of these factors affect the incidence of major errors. CONCLUSION: Contributors to journals should be aware of the various risk factors for citation errors. Citation accuracy may be improved by modifying these factors. PMID- 11430285 TI - The evaluation of the effectiveness of health education interventions in clinical practice: a continuing methodological challenge. AB - AIM: This paper examines the methodological issues arising from an evaluation of the effectiveness of a health education project undertaken to increase Hong Kong Chinese women's knowledge of the prevention of cervical cancer and the uptake of screening. BACKGROUND: The significance of health promotion to the prevention of diseases currently affecting contemporary society has become increasingly recognized. Within the context of health promotion health education continues to provide an important preventive strategy. Indeed the leading causes of mortality such as coronary heart disease and cancer lend themselves well to health education interventions. However the evaluation of the effectiveness of health education remains complex and raises some important methodological issues. DESIGN: The project used a health education intervention as the major preventive strategy and employed multiple methods of evaluation to assess its effectiveness. Outcome evaluation consisted of a confidential questionnaire administered at two points in time to measure changes in health-related behaviour and knowledge. It also included the collection of data from service providers to assess changes in the uptake of cervical cancer screening. Process evaluation involved the use of focus groups with randomly selected groups of women who had participated in the health education intervention and a diary kept by the project nurse. FINDINGS: Methodological issues identified in the evaluation of the project included the extent to which changes in health-related knowledge and behaviour could be attributed to the intervention, the sensitivity of outcome measures and challenges in developing methods of process evaluation appropriate to the target population. CONCLUSIONS: The findings highlight the complexity of designing effective evaluation strategies for health education and the need to consider these issues in the development of both process and outcome evaluation. PMID- 11430286 TI - Refinement of the concept of uncertainty. AB - AIMS: To analyse the conceptual maturity of uncertainty; to develop an expanded theoretical definition of uncertainty; to advance the concept using methods of concept refinement; and to analyse congruency with the conceptualization of uncertainty presented in the theory of hope, enduring, and suffering. BACKGROUND: Uncertainty is of concern in nursing as people experience complex life events surrounding health. In an earlier nursing study that linked the concepts of hope, enduring, and suffering into a single theoretical scheme, a state best described as 'uncertainty' arose. This study was undertaken to explore how this conceptualization fit with the scientific literature on uncertainty and to refine the concept. DESIGN/METHODS: Initially, a concept analysis using advanced methods described by Morse, Hupcey, Mitcham and colleagues was completed. The concept was determined to be partially mature. A theoretical definition was derived and techniques of concept refinement using the literature as data were applied. FINDINGS: The refined concept was found to be congruent with the concept of uncertainty that had emerged in the model of hope, enduring and suffering. CONCLUSIONS: Further investigation is needed to explore the extent of probabilistic reasoning and the effects of confidence and control on feelings of uncertainty and certainty. PMID- 11430287 TI - Informational basis for expert intuition. AB - AIM: Intuition has been cited as an integral part of nursing clinical expertise. Responding to the recent scholarly debate over the status of expert nursing intuition as part of the art or science of nursing, this article proposes an alternative view that may provide an informational basis for what has been described phenomenologically as intuition. RATIONALE: Two reasons for the dispute over the status of nursing intuition as 'art' or 'science' are proposed: The first is methodological. The second relates to intuition's close link with perception and underlying assumptions about perception. By examining intuition through an ecological psychology framework, the problem takes on a different character, one that is no longer focused solely on the expert's cognitive (or perceptual) processes, but also on the information provided by the patient and the context of care. CONCLUSIONS: This perspective has several implications for nursing. By investigating information sources (i.e. higher order variables or constraints) in the nurse-patient encounter, the problem of intuition may be clarified, and perhaps simplified. We may find that what nurse researchers have called 'intuition' is what Gibson (1966, 1986) termed 'direct perception.' Intuition as direct perception is information-based and lawful. Finally, although some aspects of intuition may be hard-wired through evolution, intuition as direct perception can be developed through education and extensive, deliberate practice with appropriate exemplars. Characterized as direct perception, intuition is an observable, lawful phenomenon that is measurable, potentially teachable, and appropriately part of nursing science. PMID- 11430288 TI - Emotions and illness. AB - AIM: This paper discusses issues central to emotional expression in illness. Its aim is to challenge the psychopathological paradigm currently utilized within healthcare and propose an alternative model based on healthy emotions. BACKGROUND: Emotions are important to human survival. It is argued that most patients will have an emotional response to their illness but anxiety and depression are unlikely to be representative of most patients experience. The paper focuses on two important negative emotions, fear and sadness, and their psychopathological associates, anxiety and depression. Problems of diagnosing anxiety and depression in practice are highlighted. It is contended that emotional reaction to illness is normal and that emotions expressed are likely to hold clues to individual adaptation. It is argued that in health care, emotions should be assessed as healthy reactions to a survival threat and merit study from this perspective if we wish to understand individual patient illness reaction and adaptation better. Literature from cardiology is used to illustrate clinical problems. RATIONALE: Given the present knowledge of emotions, it should be possible to utilize emotion theory as a paradigm for studying human reaction to illness. This in turn could help shape studies on adherence and adaptation. CONCLUSION: The reaction and experience of patient illness is unlikely to be understood through the psychopathology paradigm. A better understanding of patient reaction to their illness is more likely to emerge through the study of basic emotions during acute life-threatening episodes. PMID- 11430289 TI - Professional and organizational learning: analysing the relationship with the development of practice. AB - BACKGROUND: Organizational and professional learning are interrelated processes that underpin the contemporary drive for a quality evidence-based delivery of health care in the United Kingdom (UK). DESIGN: A soft systems methodology was used to explore the pervasiveness of practice developments. Three case study sites were identified using matrix sampling and data collected through 29 individual interviews and two focus group interviews, with the interviews augmented with a tool designed to maximize analysis of the processes of developing practice. FINDINGS: The resultant model of developing health care practice includes three processes: using and creating knowledge, understanding and practice of patient care, and effecting development. The whole model was underpinned by professional and organizational learning in which 'expert thinkers' engaged in double loop learning to reconceptualize care rather than just perpetuate existing patterns of care delivery. PMID- 11430290 TI - Emerging Statistical Issues in Public Health for the 21st Century. Proceedings of the 7th biennial CDC and ATSDR Symposium. PMID- 11430291 TI - A global network on Schistosomiasis information systems and control of snail borne diseases. Proceedings of a meeting. Bellagio, Italy, 10-14 April 2000. PMID- 11430292 TI - Reflections on distraction osteogenesis. A surgical distraction or the 'future'? PMID- 11430293 TI - Blockade of cholecystokinin-A receptors has no effect on dyskinesias in Parkinson's disease. PMID- 11430294 TI - Charles Bonnet's syndrome: complete remission of complex visual hallucinations treated by gabapentin. PMID- 11430295 TI - Paraneoplastic opsoclonus-myoclonus associated with renal cell carcinoma and responsive to tumour ablation. PMID- 11430296 TI - Azathioprine treatment in multiple sclerosis; pretreatment assessment of metaboliser status. PMID- 11430297 TI - Migrainous brain stem disturbance in Norrie disease: case report. PMID- 11430298 TI - Acute deterioration in Chiari type 1 malformation after chiropractic cervical manipulation. PMID- 11430299 TI - The EQ-5D--a generic quality of life measure--is a useful instrument to measure quality of life in patients with Parkinson's disease. PMID- 11430300 TI - Cerebral malaria. PMID- 11430301 TI - Treatment of paroxysmal sympathetic storm with labetalol. PMID- 11430302 TI - Recurrent ptosis. PMID- 11430303 TI - Management of intracranial bleeding associated with anticoagulation: balancing the risk of further bleeding against thromboembolism from prosthetic heart valves. PMID- 11430304 TI - Delirium episode as a sign of undetected dementia among community dwelling elderly subjects. PMID- 11430305 TI - AACE/AAES medical/surgical guidelines for clinical practice: management of thyroid carcinoma. American Association of Clinical Endocrinologists. American College of Endocrinology. PMID- 11430306 TI - Visual vignette. A case presentation: cerebrotendinous xanthomatosis. PMID- 11430309 TI - Multivariate modelling of polychlorinated biphenyl-induced CYP1A activity in the MH1C1 rat hepatoma cell line. PMID- 11430311 TI - Preliminary results of the ECVAM validation study on three in vitro embryotoxicity tests. PMID- 11430314 TI - Evidence based medicine and stereopsis. PMID- 11430315 TI - An apology. Re: Web correspondence. Binocul Vis Strabismus Q 2001; 16(1):11. PMID- 11430316 TI - Canadian Cardiovascular Congress 2000 and 53rd annual meeting of the Canadian Cardiovascular Society. October 29-November 1, 2000. Vancouver, British Columbia, Canada. Abstracts. PMID- 11430317 TI - Web alert. PMID- 11430318 TI - New and old threats to human health. PMID- 11430319 TI - CEA and innate immunity. PMID- 11430320 TI - O phototroph, o chemotroph, where art thou? PMID- 11430322 TI - Cumulative index 1998-2000. PMID- 11430321 TI - Will scrapie in sheep in Great Britain disappear? PMID- 11430323 TI - Abstracts of the 7th annual meeting of the Organization for Human Brain Mapping. Brighton, United Kingdom, June 10-14, 2001. PMID- 11430324 TI - Measurement of C-reactive protein for the targeting of statin therapy in the primary prevention of acute coronary events. AB - BACKGROUND: Elevated levels of C-reactive protein, even in the absence of hyperlipidemia, are associated with an increased risk of coronary events. Statin therapy reduces the level of C-reactive protein independently of its effect on lipid levels. We hypothesized that statins might prevent coronary events in persons with elevated C-reactive protein levels who did not have overt hyperlipidemia. METHODS: The level of C-reactive protein was measured at base line and after one year in 5742 participants in a five-year randomized trial of lovastatin for the primary prevention of acute coronary events. RESULTS: The rates of coronary events increased significantly with increases in the base-line levels of C-reactive protein. Lovastatin therapy reduced the C-reactive protein level by 14.8 percent (P<0.001), an effect not explained by lovastatin-induced changes in the lipid profile. As expected, lovastatin was effective in preventing coronary events in participants whose base-line ratio of total cholesterol to high-density lipoprotein (HDL) cholesterol was higher than the median ratio, regardless of the level of C-reactive protein (number needed to treat for five years to prevent 1 event, 47; P=0.005). However, lovastatin was also effective among those with a ratio of total to HDL cholesterol that was lower than the median and a C-reactive protein level higher than the median (number needed to treat, 43; P=0.02). In contrast, lovastatin was ineffective among participants with a ratio of total to HDL cholesterol and a C-reactive protein level that were both lower than the median (number needed to treat, 983; P=0.80). CONCLUSIONS: Statin therapy may be effective in the primary prevention of coronary events among subjects with relatively low lipid levels but with elevated levels of C reactive protein. PMID- 11430325 TI - Long-term effects of indomethacin prophylaxis in extremely-low-birth-weight infants. AB - BACKGROUND: The prophylactic administration of indomethacin reduces the frequency of patent ductus arteriosus and severe intraventricular hemorrhage in very-low birth-weight infants (those with birth weights below 1500 g). Whether prophylaxis with indomethacin confers any long-term benefits that outweigh the risks of drug induced reductions in renal, intestinal, and cerebral blood flow is not known. METHODS: Soon after they were born, we randomly assigned 1202 infants with birth weights of 500 to 999 g (extremely low birth weight) to receive either indomethacin (0.1 mg per kilogram of body weight) or placebo intravenously once daily for three days. The primary outcome was a composite of death, cerebral palsy, cognitive delay, deafness, and blindness at a corrected age of 18 months. Secondary long-term outcomes were hydrocephalus necessitating the placement of a shunt, seizure disorder, and microcephaly within the same time frame. Secondary short-term outcomes were patent ductus arteriosus, pulmonary hemorrhage, chronic lung disease, ultrasonographic evidence of intracranial abnormalities, necrotizing enterocolitis, and retinopathy. RESULTS: Of the 574 infants with data on the primary outcome who were assigned to prophylaxis with indomethacin, 271 (47 percent) died or survived with impairments, as compared with 261 of the 569 infants (46 percent) assigned to placebo (odds ratio, 1.1; 95 percent confidence interval, 0.8 to 1.4; P=0.61). Indomethacin reduced the incidence of patent ductus arteriosus (24 percent vs. 50 percent in the placebo group; odds ratio, 0.3; P<0.001) and of severe periventricular and intraventricular hemorrhage (9 percent vs. 13 percent in the placebo group; odds ratio, 0.6; P=0.02). No other outcomes were altered by the prophylactic administration of indomethacin. CONCLUSIONS: In extremely-low-birth-weight infants, prophylaxis with indomethacin does not improve the rate of survival without neurosensory impairment at 18 months, despite the fact that it reduces the frequency of patent ductus arteriosus and severe periventricular and intraventricular hemorrhage. PMID- 11430326 TI - Transfusion-transmitted malaria in the United States from 1963 through 1999. AB - BACKGROUND: Transfusion-transmitted malaria is uncommon in the United States. After the report of three cases of complicated Plasmodium falciparum infection acquired by transfusion, we reviewed all cases of transfusion-transmitted malaria reported to the Centers for Disease Control and Prevention (CDC) from 1963 through 1999. METHODS: Information on the patients was from surveillance reports sent to the CDC. Information about the implicated blood donors came from the National Malaria Surveillance System. To determine whether donors should have been excluded from donating blood, we compared their characteristics with the exclusion guidelines of the Food and Drug Administration and the American Association of Blood Banks. RESULTS: Of 93 cases of transfusion-transmitted malaria reported in 28 states, 33 (35 percent) were due to P. falciparum, 25 (27 percent) were due to P. vivax, 25 (27 percent) were due to P. malariae, 5 (5 percent) were due to P. ovale, 3 (3 percent) were mixed infections, and 2 (2 percent) were due to unidentified species. Ten of the 93 patients (11 percent) died. There were potentially 91 donors (in two cases, two patients received blood from the same donor), 67 of whom (74 percent) could be identified as infective. Of 64 implicated donors whose country of origin was reported, 38 (59 percent) were foreign born. Among those for whom complete information was available, 37 of 60 donors (62 percent) would have been excluded from donating according to current guidelines (in place since 1994), and 30 of 48 donors (62 percent) should have been excluded under the guidelines in place at the time of donation. CONCLUSIONS: Careful screening of donors according to the recommended exclusion guidelines remains the best way to prevent transfusion-transmitted malaria. PMID- 11430327 TI - Nephropathy and establishment of a renal reservoir of HIV type 1 during primary infection. PMID- 11430328 TI - Images in clinical medicine. Tension pneumoperitoneum. PMID- 11430329 TI - Advances in mechanical ventilation. PMID- 11430330 TI - Side effects of adjuvant treatment of breast cancer. PMID- 11430331 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 20-2001. A 64-year-old man with fever and gram-negative bacteremia. PMID- 11430332 TI - Statins and the acute-phase response. PMID- 11430333 TI - Ecology in evolution. PMID- 11430334 TI - The ecology of medical care revisited. PMID- 11430335 TI - Noninvasive ventilation in immunosuppressed patients. PMID- 11430336 TI - Noninvasive ventilation in immunosuppressed patients. PMID- 11430337 TI - Gene-expression profiles in hereditary breast cancer. PMID- 11430338 TI - Gene-expression profiles in hereditary breast cancer. PMID- 11430339 TI - Liver biopsy. PMID- 11430340 TI - Liver biopsy. PMID- 11430341 TI - Liver biopsy. PMID- 11430342 TI - Diagnosis of recurrent astrocytoma with fludeoxyglucose F18 PET scanning. PMID- 11430343 TI - From the Centers for Disease Control and Prevention. Outbreak of Legionnaires' disease among automotive plant workers--Ohio, 2001. PMID- 11430344 TI - From the Centers for Disease Control and Prevention. Deferral of routine booster doses of tetanus and diphtheria toxoids for adolescents and adults. PMID- 11430345 TI - From the Centers for Disease Control and Prevention. Update: Outbreak of acute febrile respiratory illness among college students--Acapulco, Mexico, March 2001. PMID- 11430346 TI - JAMA patient page. Breathing problems during sleep. PMID- 11430348 TI - Educational programs for deaf students. Schools and programs in Canada. PMID- 11430347 TI - The hot and the classic. PMID- 11430349 TI - Educational programs for deaf students. Postsecondary programs. PMID- 11430350 TI - University and college programs for personnel in deafness. PMID- 11430351 TI - Programs for deaf-blind children and adults. PMID- 11430352 TI - Programs and services for the deaf. Supportive and rehabilitative programs. PMID- 11430353 TI - Educational programs for deaf students. Schools and programs in the United States. PMID- 11430354 TI - A comment on codominant scoring of AFLP markers. PMID- 11430355 TI - Highly expressed genes in yeast evolve slowly. PMID- 11430356 TI - Integrated medicine. Many orthopaedic surgeons do not think of patients just as malfunctioning elbows. PMID- 11430357 TI - Integrated medicine. Integrated medicine means doctors will be in charge. PMID- 11430358 TI - Integrated medicine. Finding the time is most important. PMID- 11430360 TI - Integrated medicine. Adding complimentary medicine will not stop descent into soulless medicine. PMID- 11430359 TI - Integrated medicine. Science of the art of medicine does exist. PMID- 11430361 TI - Integrated medicine. Medicine can't be everything to everyone. PMID- 11430362 TI - Integrated medicine. Cost of consultations is key issue. PMID- 11430363 TI - Integrated medicine. Road to betrayal is short. PMID- 11430365 TI - Water fluoridation. Critical difference was overlooked. PMID- 11430366 TI - Water fluoridation. Meta-analysis of fluoridation and fractures has been done. PMID- 11430367 TI - Water fluoridation. Government should meet commitment made in white paper. PMID- 11430368 TI - Water fluoridation. Clearer evidence of benefits and risks is needed. PMID- 11430370 TI - Water fluoridation. Little is known about the safety of fluoridation. PMID- 11430369 TI - Water fluoridation. Results should be viewed with concern rather than applause. PMID- 11430371 TI - Water fluoridation. Ireland has decay in non-fluoridated communities. PMID- 11430372 TI - Pregnancy is contraindication for rubella vaccination still. PMID- 11430373 TI - Maternal age and risk of type 1 diabetes in children. Relative risks by maternal age are biased. PMID- 11430374 TI - Maternal age and risk of type 1 diabetes in children. Flawed analysis invalidates conclusions. PMID- 11430375 TI - Maternal age and risk of type 1 diabetes in children. Other secular trends may explain associations with diabetes risk. PMID- 11430376 TI - Maternal age and risk of type 1 diabetes in children. Association may disappear after adjusting for year of birth. PMID- 11430377 TI - Human consequences of foot and mouth disease are more than described. PMID- 11430378 TI - Referral guidelines for cancer need closer scrutiny. PMID- 11430379 TI - Assessment of grouping variable should have been blind in trial of dementia. PMID- 11430380 TI - Portal hypertension. Where is evidence for 5% dextrose and pulmonary artery catheter? PMID- 11430381 TI - Back to tale of seven surgeons. PMID- 11430382 TI - International Association for Dental Research divisional and sectional abstracts. PMID- 11430383 TI - XXth International Symposium on Cerebral Blood Flow, Metabolism and Function and Vth International Conference on Quantification of Brain Function with PET. Taipei, Taiwan, June 9-13, 2001. Abstracts. PMID- 11430384 TI - 8th International Symposium on Mechanisms of Vasodilatation. May 31-June 3, 2001. Boston, Massachusetts, USA. Abstracts. PMID- 11430385 TI - Premature mortality and hypopituitarism. PMID- 11430386 TI - Bartonella hensela in inflammatory bowel disease. PMID- 11430387 TI - Sudden death and hypertrophic cardiomyopathy. PMID- 11430388 TI - Ingestion of foreign objects. PMID- 11430389 TI - Normalisation of blood pressure in bleeding patients. PMID- 11430390 TI - Causes of obesity. PMID- 11430391 TI - Inhaled insulin in type 1 diabetes. PMID- 11430392 TI - Epidemiology as show business? PMID- 11430394 TI - A prospective comparison of four antihypertensive agents in daily clinical practice. PMID- 11430395 TI - Society of Cardiovascular and Interventional Radiology (SCVIR) 26th annual meeting. San Antonio, Texas, USA. March 3-8, 2001. Abstracts. PMID- 11430396 TI - Granulocyte colony-stimulating factor in a neutropenic infant with cervical lymphadenitis. PMID- 11430397 TI - Availability of case definition for acute idiopathic pulmonary hemorrhage in infants. PMID- 11430398 TI - Complexes with truncated RNAs from the large domain of Archaeoglobus fulgidus signal recognition particle. AB - Protein SRP19 is an important component of the signal recognition particle (SRP) as it promotes assembly of protein SRP54 with SRP RNA and recognizes a tetranucleotide loop. Structural features and RNA binding activities of SRP19 of the hyperthermophilic archaeon Archaeoglobus fulgidus were investigated. An updated alignment of SRP19 sequences predicted three conserved regions and two alpha-helices. With Af-SRP RNA the Af-SRP54 protein assembled into an A. fulgidus SRP which remained intact for many hours. Stable complexes were formed between Af SRP19 and truncated SRP RNAs, including a 36-residue fragment representing helix 6 of A. fulgidus SRP RNA. PMID- 11430399 TI - Kinetic regulation of an alkaline p-nitrophenylphosphate phosphatase from Halobacterium salinarum in low water system by Mn2+ and monovalent cations. AB - Reversed micelles were used as a cytoplasmic model to study the effect of the multi-ionic equilibria on kinetics of extreme halophilic enzymes. The enzymatic system used was an alkaline p-nitrophenylphosphate phosphatase from the halophilic archaeon Halobacterium salinarum (earlier halobium). This enzyme was solubilised in reversed micelles of hexadecyltrimethylammonium bromide in cyclohexane, with 1-butanol as co-surfactant. The p-nitrophenylphosphate phosphatase is a good system to study the regulation of the enzymatic activity, because it utilises manganese, water and potassium or sodium as cofactors and reacts with p-nitrophenylphosphate. Kinetic behaviour was determined by the ratio between [Mn2+] and [Na+] or [K+]. When the [Mn2+] increased and [Na+] or [K+] decreased, the kinetics showed cooperative behaviour. Rabin's model describes the kinetic behaviour of the p-nitrophenylphosphate phosphatase in reversed micelles. PMID- 11430400 TI - Transposon mutations in the flagella biosynthetic pathway of the solvent-tolerant Pseudomonas putida S12 result in a decreased expression of solvent efflux genes. AB - Fourteen solvent-sensitive transposon mutants were generated from the solvent tolerant Pseudomonas putida strain S12 by applying the TnMod-KmO mutagenesis system. These mutants were unable to grow in the presence of octanol and toluene. By cloning the region flanking the transposon insertion point a partial sequence of the interrupted genes was determined. Comparison of the deduced amino acid sequences with a protein database revealed the following interrupted putative gene products: organic solvent efflux proteins SrpA and SrpB, the flagellar structural proteins FlgK, FlaG, FliI, FliC, and FliH, the transcriptional activator FleQ, the alternative RNA polymerase sigma factor RpoN, and the flagellum-specific RNA polymerase sigma factor FliA (RpoF). The transposon mutants, except for the organic solvent efflux mutants, were nonmotile as determined by a swarm assay and the formation of the flagellum was totally impaired. Expression studies with a srp promoter probe showed a decreased expression of the SrpABC efflux pump in the nonmotile mutants. PMID- 11430401 TI - Helical growth and the curved shape of Vibrio cholerae. AB - The curved, comma, or bent shape of Vibrio cholerae is attributed to, and explained by, the normal helical growth of the cell. The comma-like shape of V. cholerae is not due to an asymmetrical positioning of peptidoglycan such that some chains of peptidoglycan are placed so they are more spread out on one side of the cell and squeezed together on the other side. PMID- 11430402 TI - Adaptation of signature-tagged mutagenesis to Escherichia coli K1 and the infant rat model of invasive disease. AB - With the exception of the polysialic acid capsule (K1 antigen), little is known about other virulence factors needed for systemic infection by Escherichia coli K1, the leading cause of Gram-negative neonatal meningitis in humans. In this work, the functional genomics method of signature-tagged mutagenesis (STM) was adapted to E. coli K1 and the infant-rat model to identify non-capsule virulence genes. Validation of the method was demonstrated by the failure to recover a reconstructed acapsular mutant from bacterial pools used to systemically infect 5 day-old rats. Three new genes required for systemic disease were identified from a total of 192 mutants screened by STM (1.56% hit rate). Gut colonization, Southern blot hybridization, mixed-challenge infection, and DNA sequence analyses showed that the attenuating defects in the mutants were associated with transposon insertions in rfaL (O antigen ligase), dsbA (thiol:disulfide oxidoreductase), and a new gene, puvA (previously unidentified virulence gene A), with no known homologues. The results indicate the ability of STM to identify novel systemic virulence factors in E. coli K1. PMID- 11430403 TI - Contribution of multidrug efflux pumps to multiple antibiotic resistance in veterinary clinical isolates of Pseudomonas aeruginosa. AB - The contribution of efflux pumps to multidrug resistance in 12 Pseudomonas aeruginosa isolates from various animal sources was assessed. Western immunoblot analyses demonstrated that all twelve isolates expressed significant levels of the MexAB OprM efflux system whereas two isolates simultaneously expressed the MexEF OprN or MexXY systems, respectively. One strain contained a single mutation in mexR, a regulator of mexAB-oprM expression, that did not adversely affect the MexR amino acid sequence, and three isolates contained the same, single base change in the mexA-mexR intergenic region. The MexXY-expressing strain contained two base substitutions in its mexZ regulatory gene which did not alter the MexR sequence. PMID- 11430404 TI - Mesophilic cyanobacteria producing thermophilic restriction endonucleases. AB - When searching for the site-specific endonucleases in several strains of Phormidium we made the following observations. Among the 16 strains that originated from 15 species of Phormidium, 12 produced one or more restriction enzymes, of which two produced the highly thermophilic restriction endonucleases PtaI and PpaAII with their optimum activity at 65-80 degrees C, which is far above the lethal temperature for the host microorganism (40 degrees C). These two temperature-resistant enzymes are isoschizomers of known BspMII and TaqI endonucleases, respectively. The presence of the thermophilic TaqI isoschizomer does not seem to play any role in the mesophilic host microorganism, which does not even contain an active cognate methyltransferase. Among the remaining 10 strains, six produced isoschizomers of endonucleases which we first described in cyanobacteria, namely: PfaAII (NdeI), PinBII and PtaI (BspMII), PlaAII (RsalI), PpaAII, PpeI (ApaI). Two enzymes, PauAII (AhaIII) and PfaAII (NdeI), belong to a group of a very rarely occurring isoschizomers. Out of 21 cyanobacterial endonucleases investigated by us, four were active in a wide range of temperatures (from 15 to 60 degrees C) which also extended the optimal growth temperature of the hosts. We assume that our observation on the presence of temperature-resistant restriction enzymes in mesophilic hosts supports the idea of horizontal gene transfer. Restriction modification systems may be an excellent tool for investigation of that phenomenon. PMID- 11430405 TI - Structures of ribonuclease P RNAs of Vibrio core species. AB - The structures of an RNA component of ribonuclease P (RNase P RNA) were examined for Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio carchariae, Vibrio natriegens, Vibrio campbellii, Vibrio proteolyticus, Vibrio pelagius and Vibrio harveyi to clearly determine their genetic differences. The RNase P RNAs ranged from 382 to 454 nucleotides (nt) in size, and were remarkably different from each other in the structure of two helices, P3 and P12. The P3 helices were comprised of tandem repeats of a palindromic sequence (24 nt), resulting in the longitudinal repetition of a stem structure. The number of repetitions ranged from four in V. harveyi, to one in both V. alginolyticus and V. proteolyticus. The genes for the RNase P RNAs of all species were located between two open reading frames, the amino acid sequences of which were similar to the hypothetical proteins located at 70.92 and 1.94 min in the Escherichia coli chromosome. PMID- 11430406 TI - Genetic relatedness of VP1 genes of Australian and Taiwanese rotavirus isolates. AB - Gene 1 (which encodes the viral RNA-dependent RNA polymerase, VP1) of an atypical human reassortant rotavirus strain, E210 (serotype G2P1B), is unrelated to genes 1 of standard human rotaviruses. To ascertain the origin of this gene, we determined a partial sequence and found that it exhibited greatest identity to gene 1 of a Taiwanese isolate, TE83, which is representative of G2 strains that caused an epidemic of gastroenteritis in 1993. Limited sequence identity to genes 1 of standard human and animal viruses was observed. This was confirmed by phylogenetic analysis. However, hybridization analysis using an E210 gene 1 specific probe indicated that a related gene was found among other Australian G2 isolates and in a Japanese strain isolated in the 1970s. PMID- 11430407 TI - CbbR, a LysR-type transcriptional regulator from Hydrogenophilus thermoluteolus, binds two cbb promoter regions. AB - The cbbR encoding the LysR-type transcriptional regulator is located downstream of cbbLSQOYA and this gene is located upstream of cbbFPT in divergent transcription. The two promoter regions with LysR-binding sites are located in the cbbL upstream region and in the cbbR-cbbF intergenic region. Electrophoretic mobility shift assays using a cell extract of Escherichia coli harboring a plasmid containing cbbR and the DNA fragments of promoter regions indicated that CbbR binds in both regions. NADPH caused differences in the complex of CbbR and DNA. PMID- 11430408 TI - IS1550 from Mycoplasma fermentans is transposable in Escherichia coli. AB - An insertion sequence (IS)-like element (ISMil) was previously isolated from the incognitus strain of Mycoplasma fermentans. With polymerase chain reaction primers corresponding to the left and right terminal inverted repeats of ISMil, a 1.4-kb DNA fragment was amplified from the genome of the M64 strain of M. fermentans. This DNA fragment has structural characteristics similar to those of ISMil and is designated IS1550. One copy of IS1550 encoded two considerable overlapping open reading frames (ORFs), ORF1 and ORF2. A putative translation frame-shift signal AAAAAAG (A6G) was located near the 3'-end of ORF1. This signal might cause a -1 frame-shift to form a fused product of ORF1 and ORF2 with 444 amino acids, which has a significant similarity to the putative transposase of the IS3 family. This copy of IS1550 was shown to be transposable in Escherichia coli ISM612. Its transposition caused a 1465-bp deletion immediately adjacent to the 3'-end of the element and the creation of a pair of 3-bp direct repeats flanking the element at the new insertion site. On the basis of these results, IS1550 was considered a typical transposable element. PMID- 11430409 TI - PCR cloning of type II polyhydroxyalkanoate biosynthesis genes from two Pseudomonas strains. AB - Two polyhydroxyalkanoate synthase genes, phaC1 from Pseudomonas pseudoalcaligenes HBQ06 and phaC2 from Pseudomonas nitroreducens 0802, were cloned using a PCR cloning strategy based on the type II pha loci property of Pseudomonas strains. The complete open reading frames (ORFs) of phaC1 (P. nitroreducens HBQ06) and phaC2 (P. nitroreducens 0802) were identified from the PCR products. Using the sequence information, the complete PHA synthase genes were PCR cloned directly from the genomic DNA and expressed in Escherichia coli as confirmed by Fourier transform-infrared spectroscopy and gas chromatography. The differences between PhaC1 and PhaC2 were analyzed and the two proteins were suggested to contain different functions and evolution history. PMID- 11430410 TI - Variant toxin B and a functional toxin A produced by Clostridium difficile C34. AB - A particular property of Clostridium difficile strain C34 is an insertion of approximately 2 kb in the tcdA-C34 gene that does not hinder expression of a fully active TcdA-C34 molecule. Intoxication with TcdA-C34 induced an arborized appearance in eukaryotic cells (D-type cytopathic effect); intoxication with TcdB C34 induced a spindle-like appearance of cells (S-type cytopathic effect). Inactivation of GTPases with purified toxins revealed that Rho, Rac, Cdc42, and Rap are substrates of TcdA-C34. The variant cytotoxin TcdB-C34 inactivated Rho, Rac, Cdc42, Rap, Ral, and R-Ras. Hence, this is the first 'S-type' cytotoxin which inactivates both Rho and R-Ras, and is coexpressed with a 'D-type' enterotoxin. Our results support the hypothesis that R-Ras is a key GTPase related to the S-type cytopathic effect and suggest that induction of a S-type cytopathic effect dominates induction of the D-type cytopathic effect. PMID- 11430411 TI - Identification of the DNA-binding protein, HrcA, of Streptococcus thermophilus. AB - HrcA is a negative transcriptional factor controlling the expression of the stress-specific operons dnaK and groESL in several bacteria. Although the HrcA structural gene has been identified in various organisms, studies at the protein level have been so far limited and mostly restricted to Bacillus subtilis. We have identified the HrcA protein of Streptococcus thermophilus and show here that it is a dimer with a native molecular mass of 74.5 kDa and a sequence-specific DNA-binding activity. Partially denatured and inactive S. thermophilus HrcA recovered its binding activity in the presence of the GroEL chaperone. PMID- 11430412 TI - The cell division genes (ftsE and X) of Aeromonas hydrophila and their relationship with opsonophagocytosis. AB - A transposon mutant from Aeromonas hydrophila AH-3 was obtained which was highly resistant to opsonophagocytosis. The mutation was identified in the ftsE gene and we characterised the operon ftsY, E and X from this bacterium. These genes, as in enteric bacteria, are neighbours to rpoH. The A. hydrophilia ftsE and X genes were fully able to complement Escherichia coli ftsE mutants, and also complement the opsonophagocytosis-resistant phenotype of the A. hydrophila mutant strain. This phenotype seems to be related to the filamentous phenotype at 37 degrees C exhibited by the A. hydrophila ftsE mutant. PMID- 11430413 TI - The MgtE Mg2+ transport protein is involved in Aeromonas hydrophila adherence. AB - Aeromonas hydrophila AH-3 strains carrying mutations in mgtE, which encodes a Mg2+ and Co2+ transport system, showed a 50% reduction of in vitro adherence to HEp-2 cells, a reduction in swarming in semisolid swarming agar, and decrease in biofilm formation of over 60% in comparison to the wild-type strain. The cloned A. hydrophila mgtE expressed from a plasmid complements a Salmonella typhimurium strain deleted for all Mg2+ transporters both phenotypically and by measurement of 57Co2+ uptake. Likewise, plasmid-borne mgtE was able to complement the changes observed in A. hydrophila mgtE mutants. We suggest that MgtE and thus Mg2+ and possibly Co2+ have a role in A. hydrophila related to their swarming ability and related consequences such as adherence and biofilm formation. PMID- 11430414 TI - Group-specific 16S rRNA targeted probes for the detection of type I and type II methanotrophs by fluorescence in situ hybridisation. AB - The study of methane-oxidising bacteria (methanotrophs) is of special interest, because of their role in the natural reduction of methane emissions from many different sources. Therefore new probes were developed to detect specifically either type I (Methylococcaceae) or type II methanotrophs (Methylocystaceae). The probes have shown high specificity in fluorescence in situ hybridisations (FISH), as demonstrated by parallel hybridisation of target and reference strains as well as sequence data analysis. With these probes, methanotrophs were detected in soil and root samples from rice microcosms, demonstrating their applicability even in a complex environmental matrix. PMID- 11430415 TI - Effects of lipids on n-alkane attenuation in media supporting oil-utilizing microorganisms from the oily Arabian Gulf coasts. AB - The Arabian Gulf is one of the most extensively oil-polluted areas of the world. The major objectives of this work were to study whether hydrocarbon-utilizing microorganisms indigenous to that area would readily accumulate added lipids, and whether this might affect their hydrocarbon consumption potential. Two prokaryotes, Arthrobacter nicotianae KCC B35 and the unidentified organisms KCC B6, as well as one eukaryote, Candida parapsilosis KCC Y1, were selected for this study. Biomass samples of the test organisms were incubated in an inorganic medium containing various concentrations of cholesterol, stearic acid, triolein or egg-phospholipids. The results revealed that all lipid classes were readily accumulated by the three test organisms. In addition, biomass samples were incubated for 6 h in an inorganic medium containing mixtures of individual lipid classes and either n-octadecane or n-docosane. The cells were removed and the residual alkanes in the medium were quantitatively recovered and analyzed by GLC. The results showed that out of the tested lipid classes only stearic acid exhibited a common stimulatory effect on the consumption of both n-alkanes by all test organisms. Other lipid classes were either inhibitory or had less pronounced effects than stearic acid. PMID- 11430416 TI - Psychopharmacology in supportive care of cancer: a review for the clinician. IV. Other psychotropic agents. AB - Besides benzodiazepine, antidepressant and neuroleptic agents, all of which have established roles in supportive care, other psychotropic drugs deserve consideration in selected conditions affecting patients with advanced cancer. This article briefly reviews relevant aspects of miscellaneous psychotropics available for secondline treatment, including nonbenzodiazepine sedative, hypnotic and anxiolytic drugs, anaesthetic agents, stimulants, and analgesic adjuvants acting on the central nervous system. The proper use of such subsidiary psychotropic agents requires that both their specificities and the particular characteristics of palliative care patients are taken into account. PMID- 11430417 TI - Clinical-practice recommendations for the management of bowel obstruction in patients with end-stage cancer. AB - The paper highlights a series of questions that doctors need to consider when faced with end-stage cancer patients with bowel obstruction: Is the patient fit for surgery? Is there a place for stenting? Is it necessary to use a venting nasogastric tube (NGT) in inoperable patients? What drugs are indicated for symptom control, what is the proper route for their administration and which can be administered in association? When should a venting gastrostomy be considered? What is the role of total parenteral nutrition (TPN) and parenteral hydration (PH)? A working group was established to review issues relating to bowel obstruction in end-stage cancer and to make recommendations for management. A steering group was established by the (multidisciplinary) Board of Directors of the European Association for Palliative Care (EAPC) to select members of the expert panel, who were required to have specific clinical and research interests relating to the topic and to have published significant papers on advanced cancer patients in the last 5 years, or to have particular clinical expertise that is recognised internationally. The final constitution of this group was approved by the Board of the EAPC. This Working Group was made up of English, French and Italian physicians involved in the field of palliative care for advanced and terminal cancer patients; and of English, American and Italian surgeons who also specialized in artificial nutrition (Dr. Bozzetti) and a professor of health economics. We applied a systematic review methodology that showed the relative lack of RCTs in this area and the importance of retrospective and clinical reports from different authors in different countries. The brief was to review published data but also to provide clinical opinion where data were lacking. The recommendations reflect specialist clinical practice in the countries represented. Each member of the group was allocated a specific question and briefed to review the literature and produce a position paper on the indications, advantages and disadvantages of each symptomatic treatment. The position papers were circulated and then debated at a meeting held in Athens and attended by all panel members. The group reviewed all the available data, discussed the evidence and discussed what practical recommendations could be derived from it. An initial outline of the results of the review and recommendations was produced. Where there were gaps in the evidence, consensus was achieved by debate. Only unanimous conclusions have been incorporated. Subsequently the recommendations were drawn together by Carla Ripamonti (Chairperson) and Robert Twycross (Co-Chair) and refined with input from all panel members. The recommendations have been endorsed by the Board of Directors of the EAPC. It was concluded that surgery should not be undertaken routinely in patients with poor prognostic criteria, such as intra abdominal carcinomatosis, poor performance status and massive ascites. A nasogastric tube should be used only as a temporary measure. Medical measures such as analgesics, anti-secretory drugs and anti-emetics should be used alone or in combination to relieve symptoms. A venting gastrostomy should be considered if drugs fail to reduce vomiting to an acceptable level. TPN should be considered only for patients who may die of starvation rather than from tumour spread. PH is sometimes indicated to correct nausea, whereas regular mouth care is the treatment of choice for dry mouth. A collaborative approach involving both surgeons and physicians can offer patients an individualized and appropriate symptom management plan. PMID- 11430418 TI - Community cancer clinics: patients' perspectives. AB - In response to the challenge of providing care for cancer patients closer to home, community chemotherapy outreach programs are being developed as part of regional cancer care organizations. The purpose of this qualitative study was to gain an understanding of cancer patients' experiences of receiving chemotherapy at community chemotherapy clinics. In total, 28 patients who had received chemotherapy at 13 community-based clinics were interviewed. Qualitative analysis of the interview transcripts revealed two prevalent themes. One theme, called "balancing gains and losses," reflected how patients considered and evaluated two dimensions when making decisions about where best to undergo treatment. The dimensions considered were quality of life versus biomedical expertise. The second theme, called "communication links," reflected patients' perceptions about the continuity of their cancer care. The results of the study suggest that the impact of the delivery of services on patients and their families needs attention. PMID- 11430419 TI - A randomized, crossover comparison of standard-dose versus low-dose lenograstim in the prophylaxis of post-chemotherapy neutropenia. AB - The aim of this trial was to compare the severity of neutropenia, the frequency of hospital admission for fever or infection, and the use of antibiotics among patients treated with a standard dose of lenograstim (263 microg/day of Euprotin) and others treated with half of this dose (131.5 microg/day) and the cost effectiveness of each of the two doses. In this single-center study, 44 patients with solid tumors, who were all receiving standard-dose chemotherapy regimens following previous neutropenia or were at high risk of neutropenia, were randomized to receive lenograstim at a dose of 263 microg or 131.5 microg daily in the first cycle and then crossed over to the alternate dose for the following cycle. Crossover to the alternate dose was repeated for patients who received more than two cycles. Lenograstim was administered from day +5 to day +14. The absolute neutrophil count (ANC) was assessed on days +5, +8, +12 and +15 of each cycle. Statistical analysis was performed using a general lineal model for repeated samples. In all, 120 cycles were administered, with a median of 3 cycles (range 1-6). Only 4 patients received only 1 cycle. No statistically significant difference (P=0.324) in ANC was observed between standard-dose (mean 5.3, 10.7, 8.3, 11.4 x 10(9)/l) and low-dose (5.0, 8.6, 5.4, 7.5 x 10(9)/l) treatment at days +5, +8, +12 and + 15. Neutropenia grade III-IV was more common in patients receiving the low than in those receiving the standard dose of lenograstim (20% vs 12%, respectively), but the difference did not reach statistical significance (P=0.1). The incidence of fever and frequency of hospital admission were not affected by the dose of lenograstim: 3 patients presented with fever with the standard dose (all of those were admitted to hospital) and 2 patients with the low dose (1 was admitted). ANC in both groups (standard and low doses) was independent of chemotherapy line (first versus second or more). Lenograstim at a dose of 131.5 microg/day is as effective as the standard dose in limiting the severity of neutropenia and in preventing episodes of fever and hospital admissions after chemotherapy for solid tumors. The lower dose of lenograstim is cost-effective in neutropenia prophylaxis. Starting its administration on day +5 reduces costs while maintaining efficacy. PMID- 11430420 TI - Oral status during radiotherapy and chemotherapy: a descriptive study of patient experiences and the occurrence of oral complications. AB - The aim of the present study was to describe patients' experiences of their oral status and their relation to oral complications during radiotherapy and chemotherapy. The sample consisted of 41 consecutive patients admitted for radiotherapy with >40 Gy including the oral cavity for tumours within the head and neck area, or for chemotherapy of haematological malignancies. At the start of radiotherapy or the second/third cycle of chemotherapy, patients rated their experiences of oral symptoms on a 100-mm visual analogue scale with endpoints "no oral discomfort" and "worst imaginable oral discomfort". Mucositis was assessed and unstimulated whole saliva was collected. Patients were examined regularly at every 10 Gy increase of radiotherapy or once a week during chemotherapy, and after completion of the radiotherapy or the chemotherapy cycle. In addition, patients given radiotherapy rated their experiences of oral symptoms 1 month after treatment. Patients receiving radiotherapy experienced a significant increase in oral symptoms over time. One month after treatment several oral symptoms were still present. The severity of mucositis increased over time. The unstimulated salivary secretion rate showed a rapid decrease after the start of radiotherapy, followed by a slow recovery. Patients who received chemotherapy reported fewer oral symptoms, with a peak 1-2 weeks after the start of treatment, and their mucositis score showed a parallel pattern of development. Their unstimulated salivary secretion rate did not change over time. There was good agreement between patient experiences and observations recorded by dental staff. PMID- 11430421 TI - Control of irinotecan-induced diarrhea by octreotide after loperamide failure. AB - Diarrhea is a well-recognized side effect of chemotherapy, which affects the quality of life and when refractory is potentially life threatening. Irinotecan (CPT-11) is associated with an elevated incidence of chemotherapy-induced diarrhea and subsequent morbidity. Standard antidiarrheal treatment is based on high-dose loperamide, but this agent is associated with a significant failure rate. Octreotide is active against chemotherapy-induced diarrhea caused by fluoropyrimidines and irinotecan, with a distinct mechanism of action. We administered octreotide in a phase I trial in 37 patients who received irinotecan and experienced loperamide-refractory diarrhea, 23 of whom experienced grade III IV diarrhea and were treated with loperamide. The 13 patients in whom to loperamide failed to control diarrhea received octreotide, with a high response rate (92%). We conclude that octreotide is effective against loperamide refractory diarrhea resulting from irinotecan-based chemotherapy. PMID- 11430422 TI - Influence of glutamine and glycyl-glutamine on in vitro lymphocyte proliferation in children with solid tumors. AB - It is reported that the proliferative response of lymphocytes is lowered in patients with solid tumors. Glutamine is a major nutrient for rapidly proliferating cells. Unlike glutamine itself, the dipeptide glycyl-glutamine as a source for glutamine is stable in aqueous solutions ex vivo. In order to evaluate the possible therapeutic role of glutamine in lymphocyte proliferation, we investigated its influence on lymphocytes of children with solid tumors before and after chemotherapy. Lymphocytes were collected from 21 children and adolescents suffering from solid tumors (before and after chemotherapy) and from healthy controls. Glutamine and glycyl-glutamine, respectively, were added to cell cultures at concentrations between zero and 1.0 mmol/l. ConA or SAC served as T- or B-cell mitogens, respectively. The lymphocyte proliferation in the healthy control group was similar in degree to lymphocyte proliferation seen in the patients with solid tumors, regardless of the mitogen used. No difference in the degree of lymphocyte proliferation before or after chemotherapy was seen with either source of glutamine. Specific subgroups of malignancies showed trends that differed from the overall findings, but these differences were not found to be statistically significant. Routine supplementation with glutamine in children with solid tumors to enhance lymphocyte function is not supported by the data gained from in vitro proliferation tests. PMID- 11430423 TI - Use of alternative/complementary therapy in breast cancer patients--a psychological perspective. AB - The objectives of this study were to assess the additional use of alternative (complementary) therapies in patients with breast cancer who were receiving conventional treatment and to compare patients using alternative therapies with patients receiving only conventional treatment with special reference to psychological adaptation, causal attribution and quality of life. A sample of 117 female out-patients with a diagnosis of breast cancer filled in the following assessment instruments: FQCI (Freiburg Questionnaire for Coping with Illness), PUK (Causal Attribution Questionnaire), EORTC QLQ-C30 (European Organization for Research and Treatment of Cancer Quality of Life Questionnaire), POMS (Profile of Mood States), and a self-developed questionnaire on alternative therapies. Nearly half the patients (47%, n = 55) reported that they had used alternative therapies in addition to conventional treatment. The methods applied most frequently were nutrition-related measures (special drinks, vitamin preparations and whole-foods each applied by about 50% of users), mistletoe preparations (49%), trace elements (47%), and homeopathy (31%). Compared with patients receiving only conventional treatment, the users of alternative therapy were younger and better educated. Users developed a more active style of illness coping than nonusers and showed more religious involvement. Patients using a large number of alternative therapies (>3) tended to adopt a more depressive coping style than those using only a small number (< or =3). For a substantial proportion of cancer patients alternative therapies apparently fulfil an important psychological need. However, a subgroup of patients using many alternative therapies seem to have considerable adjustment problems. In dealing with cancer patients the treatment team should be aware of both these groups. PMID- 11430424 TI - Electrocardiographic manifestations of heart metastasis from a primary lung cancer. AB - Though acute myocardial infarction is one of the most frequent causes of ST segment elevation, there are other, less frequent, reasons for such electrocardiographic changes. In the present case, a cardiac metastasis from a squamous cell lung carcinoma was responsible for these changes. The secondary lesion was located in the apex of the left ventricle and induced an ECG alteration mimicking myocardial ischaemia. The literature includes few reports that describe the relation between electrocardiographic changes and heart metastases, since heart metastases are not usually discovered except at autopsy. PMID- 11430425 TI - Increased Septoria musiva resistance in transgenic hybrid poplar leaves expressing a wheat oxalate oxidase gene. AB - A cDNA clone of a wheat germin-like oxalate oxidase (OxO) gene regulated by the constitutive CaMV 35S promoter was expressed in a hybrid poplar clone, Populus x euramericana ('Ogy'). Previous studies showed that OxO is likely to play an important role in several aspects of plant development, stress response, and defense against pathogens. In order to study this wheat oxalate oxidase gene in woody plants, the expression of this gene and the functions of the encoded enzyme were examined in vitro and in vivo in transgenic 'Ogy'. The enzyme activity in the transformed 'Ogy' was visualized by histochemical assays and in SDS polyacrylamide gels. It was found that the wheat OxO gene is expressed in leaves, stems, and roots of the transgenic 'Ogy' plants and the encoded enzyme is able to break down oxalic acid. Transgenic 'Ogy' leaves were more tolerant to oxalic acid as well as more effective in increasing the pH in an oxalic acid solution when compared to untransformed controls. In addition, when leaf disks from 'Ogy' plants were inoculated with conidia of the poplar pathogenic fungus Septoria musiva, which produces oxalic acid, the OxO-transformed plants were more resistant than the untransformed controls. PMID- 11430426 TI - Cloning and expression of a sorghum gene with homology to maize vp1. Its potential involvement in pre-harvest sprouting resistance. AB - Pre-harvest sprouting (PHS) in sorghum is related to the lack of a normal dormancy level during seed development and maturation. Based on previous evidence that seed dormancy in maize is controlled by the vp1 gene, we used a PCR-based approach to isolate two Sorghum bicolor genomic and cDNA clones from two genotypes exhibiting different PHS behaviour and sensitivity to abscisic acid (ABA). The two 699 amino acid predicted protein sequences differ in two residues at positions 341 (Gly or Cys within the repression domain) and 448 (Pro or Ser) and show over 80, 70 and 60% homology to maize, rice and oat VP1 proteins respectively. Expression analysis of the sorghum vp1 gene in the two lines shows a slightly higher level of vp1 mRNA in the embryos susceptible to PHS than in those resistant to PHS during embryogenesis. However, timing of expression was different between these genotypes during this developmental process. Whereas for the former the main peak of expression was observed at 20 days after pollination (DAP), the peak in the latter was found at later developmental stages when seed maturation was almost complete. Under favourable germination conditions and in the presence of fluridone (an inhibitor of ABA biosynthesis), sorghum vp1 mRNA showed to be consistently correlated with sensitivity to ABA but not with ABA content and dormancy. PMID- 11430427 TI - Changes in gene expression during programmed cell death in tomato cell suspensions. AB - To identify genes involved in plant programmed cell death (PCD), changes in gene expression during PCD in a model system of suspension-cultured tomato cells were studied. In this system, cell death is triggered by treatment with camptothecin, an inhibitor of topoisomerase 1. Cell death was accompanied by internucleosomal DNA degradation, indicating that the cell death process shares similarities with apoptosis in animals. Tomato homologues of DAD1 and HSR203, two genes that have been implicated in PCD, were isolated. During camptothecin-induced PCD tomato DAD1 mRNA levels roughly halve, while tomato HSR203 mRNA levels increase 5-fold. A differential display approach was used to identify novel genes that show changes in expression levels during camptothecin-induced PCD. This resulted in isolation of two up-regulated (CTU1 and CTU2) and four down-regulated (CTD1, CTD2, CTD4, and CTD5) cDNA clones. CTU1 shows high homology to various gluthatione S-transferases, whereas CTU2 is as yet unidentified. CTD1 is highly similar to Aux/IAA early-auxin-responsive genes. CTD2 corresponds to the tomato RSI-I gene, CTD4 is an unknown clone, and CTD5 shows limited homology with a proline-rich protein from maize. Addition of the calcium channel blocker lanthanum chloride prevented camptothecin-induced cell death. The effect of lanthanum chloride on camptothecin-induced gene expression was studied to discriminate between putative cell death genes and general stress genes. The possible role of the various predicted gene products in plant PCD is discussed. PMID- 11430428 TI - Effects of the polyubiquitin gene Ubi. U4 leader intron and first ubiquitin monomer on reporter gene expression in Nicotiana tabacum. AB - We have previously shown by RNA gel blot analyses that the tobacco polyubiquitin encoding gene Ubi.U4 is expressed in a complex pattern during plant development (Genschik et al., 1994). In order to study its tissue-specific expression, we cloned the fragment containing the -263 bp proximal promoter of the gene, the leader intron and the first ubiquitin monomer in front of the reporter GUS gene. Histochemical analyses for GUS activity during tobacco plant development revealed that the gene is expressed at variable amounts in many plant tissues with high levels in metabolically active and/or dividing cells and in the vascular tissues of the plant. We also analysed the expression pattern of constructs in which either the intron or the intron together with the first ubiquitin monomer were deleted. Our results indicate that the ubiquitin leader intron is not only a quantitative determinant of gene expression but may also influence the tissue specific expression pattern. PMID- 11430429 TI - Indications for post-translational regulation of Vitis vinifera L. arginine decarboxylase. AB - In order to study arginine decarboxylase regulation, we produced an antiserum against a hybrid of a 615 amino acid residue fragment of grapevine arginine decarboxylase cDNA with maltose-binding protein. The antiserum generated recognized mainly a protein band of ca. 80 kDa in extracts from grapevine tissues. Extracts from leaves and internodes in different developmental stages showed differences in the quantity of the 80 kDa band recognized by the antiserum. However, these differences did not correspond with changes in arginine decarboxylase specific activity. Furthermore, western blot analysis of extracts from cell cultures, where enzyme-specific activity was induced or repressed, did not reveal respective changes in the quantity of the 80 kDa protein band. Digestion of the hybrid by the specific protease factor Xa resulted in a polypeptide of 90 kDa instead of the expected two polypeptides of 43 and 66 kDa. Finally, western blot analysis of shoot extract incubated with factor Xa or the hybrid protein previously digested by factor Xa revealed that factor Xa-digested hybrid protein cleaved the 80 kDa band, resulting in two bands of ca. 38 and 40 kDa, whereas factor Xa alone did not affect it. These results suggest that arginine decarboxylase protein levels and/or activity is post-translationally regulated, as has been shown for other enzymes of polyamine biosynthesis. PMID- 11430430 TI - Temporal and tissue-specific expression of the tobacco ntf4 MAP kinase. AB - The large number of mitogen-activated protein (MAP) kinase genes identified to date in plants suggests that their encoded proteins have a wide array of functions in development and physiological responses, as has been indicated by studies on the factors which lead to the activation of these kinases. Signalling pathways involving members of a multigene family employ a variety of mechanisms to ensure response specificity, one of which is via differential gene expression. We have performed detailed analyses of the expression of the tobacco ntf4 MAP kinase gene using a variety of approaches. The ntf4 gene promoter region was isolated and a chimeric ntf4 promoter-GUS fusion construct was introduced into plants. GUS expression was detected in pollen, in developing and mature embryos, and shortly after seed germination, but not in other floral tissues and tissues such as leaf, root, or stem. This expression pattern was confirmed by northern and western analyses. In situ hybridization and immunolocalization studies showed that the expression of the ntf4 gene and its encoded protein p45Ntf4 occurred in embryos at least from the globular embryo stage until the mature seed, as well as in the seed endosperm. Taken together, the results show that the p45Ntf4 MAP kinase has a very restricted expression pattern, being found only in pollen and seeds. These findings should be important when considering MAP kinase function in plants. PMID- 11430431 TI - Survey of gene expression in winter rye during changes in growth temperature, irradiance or excitation pressure. AB - Previous comparisons of winter rye plants (Secale cereale L. cv. Musketeer) grown in a combination of specific temperature (degrees C)/irradiance (micromol m(-2) s(-1)) regimes (20/50; 20/250; 20/800; 5/50; 5/250) revealed (1) that photosynthetic acclimation to low temperature mimics photosynthetic acclimation to high light because both conditions result in comparable reduction states of photosystem II (PSII), that is, comparable PSII excitation pressure; (2) that the relative redox state of PSII also appears to regulate a specific cold acclimation gene, Wcs19. In order to identify additional genes regulated differentially by either low temperature, irradiance or excitation pressure, we initiated a detailed analysis of gene expression. We identified and characterized 42 differentially expressed genes from wheat and rye. Based on their patterns of regulation under the five growth conditions employed, 37 of the cDNAs could be classified into four groups: genes regulated by PSII excitation pressure, low temperature, growth irradiance and interaction between growth temperature and irradiance. Partial sequence analyses revealed that several of these genes encode known chloroplastic proteins such as ELIPs, transketolase, carbonic anhydrase and Mg-chelatase. However, five of the genes could not be classified unambiguously into any one of these four categories. The implications of these results and the limitations of the experimental design are discussed in terms of larger-scale genomic studies designed to understand the interactions of multiple abiotic stresses to which a plant may be exposed when examining regulation of gene expression. PMID- 11430432 TI - Partial characterization of genes whose transcripts accumulate preferentially in cell clusters at the earliest stage of carrot somatic embryogenesis. AB - We attempted to identify genes that are preferentially expressed immediately after somatic cells divide to form cell clusters at the earliest stage of carrot somatic embryogenesis when they are not or barely expressed in non-embryogenic suspension-cultured cells in the presence of 2,4-D. Using the differential display technique, we isolated three cDNA clones, designated No. 43, No. 87 and No. 93. The No. 43 transcript was preferentially expressed in the earliest cell clusters, its level decreased drastically at the globular and heart-shaped and torpedo-shaped stages, and it was not detected in non-embryogenic suspension cultured cells. No. 43 cDNA encoded a protein with homology to thaumatin-like proteins and the deduced positions of seven cysteine residues in the 63 amino acid sequence from the carboxyl terminus were identical to those in thaumatin like proteins. The full-length nucleotide sequence of No. 93 cDNA was determined and its product was about 80% homologous to precursor of the 14 kDa proline-rich DC 2.15 protein of carrot at the amino acid level. However, the deduced amino acid sequence lacked the characteristic core of repeating Pro-X motifs found in DC 2.15. The No. 93 transcript accumulated preferentially in the earliest cell clusters but it was also detected at a low level in non-embryogenic suspension cultured cells, unlike DC 2.15 transcripts that begin to accumulate in heart shaped embryos before their level falls in torpedo-shaped embryos. No. 87 transcripts were expressed preferentially in the earliest cell clusters that has been incubated with 2,4-D but were also detected at a low level in suspension cultured cells subcultured in the continued presence of 2,4-D. The No. 87 cDNA exhibited no significant homology to any sequences in databases. PMID- 11430433 TI - Tr288, a rehydrin with a dehydrin twist. AB - The rehydrin Tr288, originally isolated from a screen for differentially expressed transcripts during rehydration of desiccated moss (Tortula ruralis), was further characterized. Steady-state mRNA levels for Tr288 increase dramatically during slow drying even though protein synthesis is completely inhibited during this process. Tr288 transcripts do not accumulate during rapid drying of moss gametophytes. Conversely, during rehydration of rapidly dried tissue Tr288 transcript levels increase several-fold, while the relatively high amount of Tr288 mRNA sequestered in slowly dried material declines with time after the addition of water. Steady-state Tr288 mRNA also increases after treatment with salt (NaCl) and elevated temperature (37 degrees C). Genomic Southern analysis and isolation of a genomic clone suggest the presence of a single Tr288 gene containing two introns within the T. ruralis genome. The only sequence homology detected by a BLAST search of GenBank occurred at the 3' end of the Tr288 coding region and indicated a single copy of the K segment common to dehydrins. Computer translation of the Tr288 coding region revealed 15 copies of a protein segment (the GPN segment) that is predicted to form amphipathic alpha helices. PMID- 11430434 TI - Alpha-tubulin genes are differentially expressed during leaf cell development in barley (Hordeum vulgare L.). AB - Intricate changes in the patterns of the cytoskeleton, especially of microtubules, appear to control the establishment of complex plant cell shapes. Little is known about how these changes are accomplished. The objective of the present study was to test whether or not alpha-tubulin genes are differentially expressed during cell shaping in growing leaves of barley (Hordeum vulgare L.). Five alpha-tubulin genes representing at least most members of the gene family were found to be expressed in the leaf. Dot-blot analyses revealed expression patterns that could be classified into three groups. Two isotypes (HVATUB2 and HVATUB4) were maximally expressed in the meristem with a steady decline during the differentiation process (1). One isotype (HVATUB3) appeared to be constitutively expressed during cell shaping, although strongest signals were found during late stages, before the general decline in microtubular activity (2). The most striking finding was that two types (HVATUB1 and HVATUB5) were almost exclusively expressed in early post-mitotic cells, when transverse microtubular bundles determining the future cell shape in the mesophyll are formed (3). Relative transcript abundance was highest in HVATUB2 and HVATUB3, whereas the transcript level of the only transiently expressed HVATUB5 was very low, even during its phase of maximum expression. The results are discussed in the context of the general debate relating to the significance of multiple tubulin isotypes. PMID- 11430435 TI - Cloning of DNA encoding a catalytic subunit of SNF1-related protein kinase-1 (SnRK1-alpha1), and immunological analysis of multiple forms of the kinase, in spinach leaf. AB - Using a PCR approach, we have cloned DNA encoding a catalytic subunit isoform (SnRK1-alpha1) of SNF1-related protein kinase-1 from spinach leaf. The predicted amino acid sequence falls into the SnRK1a sub-family, and is closely related to SnRK1a sequences expressed in cucumber, Arabidopsis thaliana, tobacco and potato. We have generated two affinity-purified antipeptide antibodies (anti-RASS and anti-AEF) based on the predicted amino acid sequence of spinach SnRK1-alpha1. They were used to analyse multiple forms of SNF1-related kinase (HRK-A, -C, -D) that were previously identified by biochemical criteria in extracts of spinach leaf (Sugden et al., Plant Physiol. 120 (1999), 257-274). Anti-AEF appears to be specific for the SnRK1-alpha1 isoform, whereas anti-RASS is a 'pan-alpha' antibody that precipitates all isoforms present in spinach leaf extracts. The activities of HRK-A and HRK-C can be entirely accounted for by the SnRK1-alpha1 catalytic subunit. By contrast, only a small proportion of HRK-D activity (ca. 20%) can be accounted for by SnRK1-alpha1, with the remainder presumably being due to other isoforms (SnRK1-alpha2?) that are currently poorly defined. A 35 kDa polypeptide recognized by an antibody against the putative Arabidopsis beta2 subunit co-precipitates with HRK-C, but not HRK-A or D. PMID- 11430436 TI - Laser-mediated cartilage reshaping with feedback-controlled cryogen spray cooling: biophysical properties and viability. AB - BACKGROUND AND OBJECTIVE: Recent studies have indicated that chondrocyte viability decreases with prolonged or repeated laser irradiation. To optimize laser-mediated cartilage reshaping, the heating process must be finely controlled. In this study, we use high-power Nd:YAG laser irradiation (lambda = 1.32 microm) combined with cryogen spray cooling (CSC) in an attempt to reshape porcine septal cartilage while enhancing chondrocyte viability. STUDY DESIGN/MATERIALS AND METHODS: Chondrocyte viability was determined after high power (50 W/cm2) Nd:YAG-mediated cartilage reshaping with and without cryogen spray cooling (CSC) and correlated with dynamic measurements of tissue optical and thermal properties. RESULTS: After 1.5 to 2.0 seconds of laser exposure, characteristic changes in diffuse reflectance (indicating the onset of accelerated stress relaxation) was observed in both laser only and laser with CSC specimens. After 2 seconds of laser exposure, specimens in both groups retained the curved shape for up to 14 days. After one laser exposure, chondrocyte viability was 94.35 +/- 6.1% with CSC and 68.77 +/- 20.1% (P < 0.05) without CSC. After two laser exposures, a similar trend was observed with CSC (70.18 +/- 16.44%) opposed to without CSC (28 +/- 45%; P < 0.05). CONCLUSION: CSC during high-power laser irradiation allows rapid heating while minimizing extreme front surface temperature elevations and axial thermal gradients. Laser irradiation with CSC can be used to effectively reshape cartilage tissue with the additional advantage of increasing chondrocyte viability. PMID- 11430437 TI - Experimental and clinical results of Er:YAG laser stapedotomy. AB - BACKGROUND AND OBJECTIVE: At the beginning of the 1980s, different laser types were used for stapes surgery to reduce potential harm to inner ear structures through manipulation with conventional instruments during stapedotomy. Most clinical studies were carried out with the CO2 or the argon laser. The Er:YAG laser has been used rarely in patients with otosclerosis. STUDY DESIGN/MATERIALS AND METHODS: In an experimental study on 54 human petrous bones, the optimal laser energy parameter for dissection of the posterior stapes crus and the footplate perforation were determined. With these parameters, stapedotomy was carried out with the Er:YAG laser in 29 patients with otosclerosis with a conventional dissection of the incudostapedial joint and the stapedius muscle tendon. The Er:YAG laser was used (60 or 100 mJ, 3-6 pulses) for dissection of the posterior stapes crus and footplate perforation. RESULTS: No intra- or postoperative complications were observed in all 29 patients. Vertigo and hearing loss were not observed intra- or postoperatively. The postoperative hearing results (improvement of the air-bone gap) was in all cases satisfactory (median remaining air bone gap, 8.1 dB). The median operation time was 29 minutes (15-42 minutes) and did not show a significant prolongation in comparison to the conventional technique. In 1 of the 29 patients, the footplate perforation needed to be carried out conventionally. CONCLUSION: For the first time, Er:YAG laser parameters have been optimized and refined in a human petrous bone model and were then used in a clinical setting. According to the presented results, the Er:YAG laser seemed to be a very suitable instrument for stapedotomy. PMID- 11430438 TI - Lightdosimetric quantitative analysis of the human petrous bone: experimental study for laser irradiation of the cochlea. AB - BACKGROUND AND OBJECTIVE: Application of laser irradiation targeting the inner ear has to be investigated for therapeutic effectiveness in cochlear injury and dysfunction. In vitro data demonstrate low-level laser-induced photochemical and photobiologic cell response, depending on cell type and irradiation parameters such as light dose. The aim of the presented study was to determine the light dose received by the cochlear hair cells by using different irradiation modalities for the human petrous bone. STUDY DESIGN/MATERIALS AND METHODS: Lightdosimetric assessment was performed in human cadaver temporal bones (n = 13) after removing the cochlear membranous labyrinth. The external auditory meatus, the tympanic membrane (quadrants), and the mastoid bone were illuminated by a helium-neon laser (lambda = 593 nm) and diode lasers of different wavelengths (lambda = 635, 690, 780, and 830 nm). The spatial distribution of transmitted light in the cochlear windings was measured by means of a retrocochlearly positioned endoscopic CCD camera for image processing and was assigned to acoustic frequencies according to the tonotopic organization of the cochlea. For an estimation of the corresponding space irradiance in an intact cochlea, correction factors have been calculated by a Monte Carlo procedure on the basis of experimentally determined optical properties of skull bone. RESULTS: The transmission of light across the tympanic cavity and the promontory depends strongly on wave-length of the laser and the position of the radiator. Transtympanal irradiation results in spatial intensity variations of a factor 4 to 10 within the cochlear windings. The space irradiance in an intact cochlea is 10 to 20 times the measured irradiance. For an irradiation of the mastoid, the light transmission within the cochlea is 10(3) to 10(5) times smaller compared with an irradiation of the tympanic membrane and is extremely variable for different specimens. CONCLUSION: The strong dependence of the cochlear light distribution on various irradiation parameters demonstrates the impact of preclinical lightdosimetric investigations for effective individual laser irradiation of the human cochlea. Because of the observed spatial intensity variations, the optimal external light dose has to be chosen with regard to the tonotopy of the human cochlea. The obtained results are enabling us to apply defined laser light doses to different cochlear winding areas. Mastoidal irradiation leads to therapeutically insufficient light doses within reasonable treatment times, whereas transmeatal irradiation is recommendable. Further studies are mandatory for development of clinical devices for transmeatal irradiation of the cochlea. PMID- 11430439 TI - Low intensity laser therapy (830nm) in the management of minor postsurgical wounds: a controlled clinical study. AB - BACKGROUND AND OBJECTIVE: The stimulatory effects of low intensity laser therapy (LILT) have been widely published for the treatment of chronic ulceration. In contrast to this previous work, the current study investigated its potential efficacy (by using a dosage of 9 J/cm2) in the management of acute wounds. For this purpose, uncomplicated postoperative wounds after minor podiatric surgery were examined. STUDY DESIGN/MATERIALS AND METHODS: The study was designed as a controlled group study. Ethical approval was granted by the University of Ulster's Research Ethics Committee. Patients (n = 9) presenting with a total of 12 wounds after minor surgical procedures (partial/total nail avulsions/electrosurgery) were recruited from the Podiatry Teaching Clinic, Northern Ireland. Patients attended the clinic once per week for assessment and treatment. Weekly irradiation was performed by using a CBM Master 3 (CB Medico, Copenhagen, Denmark) diode laser (GaAlAs). The physical parameters of the output of this unit were as follows: wavelength, 830 nm; average power output, 30 mW; spot size, 0.1 cm2; irradiance, 300 mW/cm2; continuous wave output. Wound assessment and recording of pain levels were conducted weekly. Wound measurement was completed by using planimetry and digitising methods. RESULTS: Current findings indicated no statistically significant differences between Laser and Control groups for wound closure (P = 0.28 digitising; P = 0.49 planimetry) nor for pain levels reported (P = 0.88). CONCLUSION: It would seem that LILT provides no advantages in the management of minor postoperative wounds over current practice. Despite no apparent benefit of infrared laser at this dosage in the management of acute stage wounds, further research is required to determine its potential efficacy in the management of other wound types. PMID- 11430440 TI - Low intensity monochromatic infrared therapy: a preliminary study of the effects of a novel treatment unit upon experimental muscle soreness. AB - BACKGROUND AND OBJECTIVE: The current study (for which ethical approval was obtained) sought to assess the effect of low intensity monochromatic infrared therapy (LIMIRT) on experimentally induced delayed onset muscle soreness (DOMS). STUDY DESIGN/MATERIALS AND METHODS: Healthy volunteers were recruited (n = 24, 12M:12F) and randomly allocated under strict double blind conditions to one of three experimental groups (n = 8, 4M:4F): Control, Placebo, or Treatment (840 nm; 3.0 J cm(-2), pulse frequency 1 kHz). DOMS was induced in a standardised manner in the biceps brachii of the nondominant arm. Subjects attended on 5 consecutive days during which the degree of pain and functional impairment was assessed. RESULTS: Analysis of results by using nonparametric Freidman and Kruskal-Wallis H tests (with relevant post hoc tests) revealed significant differences (P < 0.05) between Control and LIMIRT treatment groups for pain and tenderness scores. Despite trends in favour of the Treatment group, analysis failed to show any significant differences between the LIMIRT treatment and Placebo groups for all variables except mechanical pain threshold points 3-6 on day 2. CONCLUSION: The results of the current study suggest that LIMIRT is ineffective in the management of DOMS at the parameters investigated. PMID- 11430441 TI - Retrospective clinical comparison of hemangioma treatment by flashlamp-pumped (585 nm) and frequency-doubled Nd:YAG (532 nm) lasers. AB - BACKGROUND AND OBJECTIVE: Laser therapy using the flashlamp-pumped pulsed dye laser (FPDL), along with contact cryotherapy, has proven to be an effective method with few side effects in the early treatment of superficial hemangiomas in infancy. The long-pulsed frequency-doubled Nd:YAG laser (VersaPulse) is a new kind of laser whose efficacy and safety we examined in direct comparison with the FPDL. STUDY DESIGN/MATERIALS AND METHODS: In a retrospective comparative study, we treated 50 infants with a total of 62 superficial hemangiomas. A total of 29 hemangiomas were treated with the FPDL (wavelength 585 nm, impulse duration 0.3 0.45 msec), 33 with the VersaPulse (wavelength 532 nm, impulse duration 1-50 msec). With an average of 3.0 and 2.6 treatments (FPDL/VersaPulse respectively), a cessation of growth or regressive tendency could be achieved in 93% and 70% of the hemangiomas, respectively. Complete regression occurred in 41% (FPDL) or 30% (VersaPulse). Only in 7% (FPDL) or 18% (VersaPulse) was there progression despite laser therapy. The side effect rates of both laser systems were low. CONCLUSION: The FPDL has proven to be the slightly more effective treatment method in comparison and, therefore, remains the therapy of choice for the initial treatment. If parents wish to pursue further treatment in the regression phase of hemangiomas to optimize the cosmetic appearance, we recommend the VersaPulse due to its low rates of side effects and painfulness. PMID- 11430442 TI - Impact of solubility on laser tissue-welding with albumin solid solders. AB - BACKGROUND AND OBJECTIVE: The correlation between the solubility of solid albumin solders and their laser weld strength was investigated. STUDY DESIGN/MATERIALS AND METHODS: Sections of dog intestine were laser welded with soluble or insoluble solid strips of solder. Two different treatments were followed for tissue soldering: "wet weld" and "dry weld." These treatments were chosen to assess the impact of solubility on the repair strength. The laser power and radiation dose were 0.14 W and 14 J/mg, respectively. Calorimetric measures of solders were also performed. RESULTS: The moisture on the tissue partially dissolved the soluble strips at the tissue interface. Hence, the repair strength of the soluble solder was significantly stronger than the repair strength of the insoluble solder (0.22 N and 0.06 N, P < 0.0001). Temperature (approximately 70 degrees C) and enthalpy variation (approximately 1.4 J/g) for denaturing the soluble and insoluble solders were not significantly different (P > 0.05). CONCLUSION: The soluble solid solder behaved like dense liquid solder at the tissue interface. Hence, the interface strength of these two forms of solder should be similar. This correlation made it possible to identify an intrinsic limit for the weld strength of albumin solders. PMID- 11430443 TI - Carotid artery anastomosis with albumin solder and near infrared lasers: a comparative study. AB - BACKGROUND AND OBJECTIVE: Laser tissue-welding has been used for anastomosis of carotid arteries. During welding, thermal injury sustained by the vessel walls should be minimized to prevent thrombosis. Two different types of lasers were used and effects on tissue damage were studied in vitro and in vivo. STUDY DESIGN/MATERIALS AND METHODS: End-to-end anastomosis of dog carotid arteries (n = 10) was performed by using a human albumin solder (HAS) in conjunction with Nd:YAG or diode lasers (lambda = 1.32 microm and 1.9 microm, respectively). The arteries were evaluated for patency and evidence of histologic injury after 21 days. Another group of arteries was laser soldered in vitro to measure the intimal and adventitial temperatures by using thermocouples. RESULTS: The arteries repaired with the diode laser sustained significantly less thermal damage than those repaired with Nd:YAG laser, both in vitro and in vivo. In particular, the intimal temperature was significantly lower (P < 0.05) for the diode than for the Nd:YAG repairs (approximately 35 degrees C and approximately 50 degrees C, respectively). In the latter group, the patency rate was 75%, but thrombosis occurred in 75% of the specimens at 21 days. All diode anastomoses were patent and thrombosis developed in only 17% of the arteries. CONCLUSION: Use of the diode laser and albumin solders may provide a means to successfully repair carotid arteries with minimal thermal damage. PMID- 11430444 TI - Reflectance spectroscopy for in vivo characterization of ovarian tissue. AB - BACKGROUND AND OBJECTIVE: To explore whether reflectance spectroscopy can differentiate normal ovary, benign neoplasms, and ovarian cancer. STUDY DESIGN/MATERIALS AND METHODS: Reflectance spectra (390-600 nm) were measured at three source-detector separations (SDS) in vivo at 64 sites in 16 patients undergoing oophorectomy. Parameters with largest statistical differences were identified. Based on these parameters algorithms were developed and evaluated. RESULTS: Promising parameters were the reflectance intensity from 540 to 580 nm (SDS, 1.1 mm), the slope of the reflectance spectrum from 490 to 520 nm (SDS, 1.1 mm), the slope from 510 to 530 nm (SDS, 2.1 mm), and the slope from 510 to 530 (SDS, 3 mm). Average sensitivity and specificity were 86 +/- 6% and 79 +/- 5% to separate normal ovary from benign neoplasms and cancers. Average sensitivity and specificity were 86 +/- 4% and 80 +/- 8% to separate ovarian cancers from benign neoplasms and normal ovary. CONCLUSION: Reflectance spectroscopy should be further investigated for ovarian cancer screening. PMID- 11430445 TI - Spectroscopic characterisation of carotid atherosclerotic plaque by laser induced fluorescence. AB - BACKGROUND AND OBJECTIVE: An effort has been made to distinguish the composition of carotid atherosclerotic plaques (CAP) from patients undergoing carotid endarterectomy, by laser induced fluorescence spectroscopy. STUDY DESIGN/MATERIALS AND METHODS: Different excitation wavelengths were used: 476, 488, and 458 nm of a continuous wave krypton/argon ion laser, and 337 nm of a pulsed nitrogen laser. Twenty-three CAP samples from different patients were investigated and several spectra from each plaque were obtained. RESULTS: Results were crossed-examined with conventional histologic techniques, which showed three areas of different composition on the pathologic samples: fibrous tissue, lipid constituents, and calcified plaque. Gaussian fittings were performed to reproduce the fluorescence spectra as a correlation of multiple Gaussian curves. CONCLUSION: The accuracy for discrimination of the heterogeneous composition of the atherosclerotic plaque is still limited, due to superposition of the fluorescence emission of various plaque components. PMID- 11430446 TI - Myofibroblasts in healing laser excision wounds. AB - BACKGROUND AND OBJECTIVE: The lack of myofibroblasts, cells responsible for wound contraction, has been suggested to be the underlying factor to the clinically observed minimal contraction in CO2 laser wounds. However, the histologic background to this phenomenon in laser excisions has not been thoroughly clarified. Therefore, we analyzed the expression of myofibroblasts in healing laser excisions and control excisions made by scalpel. STUDY DESIGN/MATERIALS AND METHODS: CO2 laser (continuous wave, 5 W) or scalpel excision wounds were created in the dorsal tongue mucosa of 144 rats. Sixteen additional rats were kept as untreated controls. Specimens from the tongues were cut at 16 different healing time points and fixed in 10% formalin. Immunohistochemical stainings with monoclonal antibodies to vimentin and to alpha-smooth muscle actin were done to determine microscopically the contractile type of myofibroblasts. RESULTS: The maximum amount of myofibroblasts was almost three times higher in scalpel than in laser excisions. The peak value was reached at 4 days in laser and at 3 days in scalpel wounds. The increase reverted to normal levels at 14 days in laser and at 6 days in scalpel wounds, respectively. CONCLUSION: Myofibroblasts appeared and disappeared slower in laser wounds. There were clearly fewer myofibroblasts in CO2 laser than in corresponding scalpel excisions known to heal by contraction. The lack of contractile myofibroblasts, therefore, is suggested as the reason for the minimal degree of contraction in CO2 laser excision wounds. PMID- 11430447 TI - Metabolic activity and DNA integrity in human hepatic metastases after interstitial laser coagulation (ILC). AB - BACKGROUND AND OBJECTIVE: For investigations into interstitial laser coagulation (ILC) of solid tumors, tissue whitening is used as a parameter for the extent of coagulation. This obvious demarcation is associated with global thermal denaturation, but it is not clear whether this finding is a good indicator of the exact outer boundary of the lethal tissue effect. STUDY DESIGN/MATERIALS AND METHODS: ILC with portal inflow occlusion was performed in human hepatic metastases of colorectal carcinoma directly after surgical resection (n = 5) or before surgical resection (n = 5) with laser parameters adapted to tumor diameter. Mitochondrial NADH-diaphorase activity and DNA integrity were assessed by histoenzymatic staining. RESULTS: In 7 of 10 tumors (mean diameter, 3.7 cm), an area of macroscopic coagulation (mean diameter, 4.2 cm) encircled the tumor in all three axes. Macroscopic coagulation corresponded to absent metabolism and disintegrated DNA. Furthermore, the macroscopic volumes of coagulation produced in tumor were comparable to the dimensions in normal porcine liver with the same laser parameters. CONCLUSION: ILC with portal inflow occlusion results in areas with complete cell avitality in the zone of tissue whitening in human hepatic liver metastases. PMID- 11430448 TI - NIAID announces plan to battle leading killer diseases. PMID- 11430449 TI - Osteoporosis in men. PMID- 11430450 TI - Methadone maintenance. PMID- 11430451 TI - Medical surveillance: the role of the family physician. PMID- 11430452 TI - Please don't call me 'provider'. PMID- 11430453 TI - HAIR-AN syndrome: a multisystem challenge. AB - HAIR-AN syndrome is an acronym for an unusual multisystem disorder in women that consists of hyperandrogenism (HA), insulin resistance (IR) and acanthosis nigricans (AN). The precipitating abnormality is thought to be insulin resistance, with a secondary increase in insulin levels and subsequent overproduction of androgens in the ovaries. Long periods of hyperinsulinism and, some suspect, hyperandrogenism can result in the cutaneous manifestation of acanthosis nigricans. Patients are often concerned about the physical manifestations of this disorder, including virilization and acanthosis nigricans, and may be less aware of systemic problems. Physicians should assess women with these problems for an underlying endocrine abnormality. Although a treatment regimen for the HAIR-AN syndrome has not been established, antiandrogen therapy and weight loss are useful. PMID- 11430454 TI - Common anorectal conditions: Part I. Symptoms and complaints. AB - Anorectal symptoms and complaints are common and may be caused by a wide spectrum of conditions. Although most conditions are benign and may be successfully treated by primary care practitioners, a high index of suspicion for colorectal cancer should be maintained, and all patients should be appropriately investigated. Inspection, palpation and anoscopic examination using an Ive's slotted anoscope provide adequate initial assessment. Pruritus ani usually represents a self-perpetuating itch-scratch cycle and is uncommonly due to infection. The history, as well as the physical examination, can distinguish anal pain due to hemorrhoids, fissure, abscess, cancer or proctalgia fugax. The most frequent causes of rectal bleeding are hemorrhoids, fissures and polyps. Diagnoses associated with difficulty in passing stool can range from constipation to fecal incontinence. PMID- 11430455 TI - Methadone therapy for opioid dependence. AB - The 1999 Federal regulations extend the treatment options of methadone-maintained opioid-dependent patients from specialized clinics to office-based opioid therapy (OBOT). OBOT allows primary care physicians to coordinate methadone therapy in this group with ongoing medical care. This patient group tends to be poorly understood and underserved. Methadone maintenance therapy is the most widely known and well-researched treatment for opioid dependency. Goals of therapy are to prevent abstinence syndrome, reduce narcotic cravings and block the euphoric effects of illicit opioid use. In the first phase of methadone treatment, appropriately selected patients are tapered to adequate steady-state dosing. Once they are stabilized on a satisfactory dosage, it is often possible to address their other chronic medical and psychiatric conditions. The maintenance phase can be used as a long-term therapy until the patient demonstrates the qualities required for successful detoxification. Patients who abuse narcotics have an increased risk for human immunodeficiency virus infection, hepatitis, tuberculosis and other conditions contributing to increased morbidity and mortality. Short- or long-term pain management problems and surgical needs are also common concerns in opioid-dependent patients and are generally treatable in conjunction with methadone maintenance. PMID- 11430456 TI - Diagnosis and management of osteomyelitis. AB - Acute osteomyelitis is the clinical term for a new infection in bone. This infection occurs predominantly in children and is often seeded hematogenously. In adults, osteomyelitis is usually a subacute or chronic infection that develops secondary to an open injury to bone and surrounding soft tissue. The specific organism isolated in bacterial osteomyelitis is often associated with the age of the patient or a common clinical scenario (i.e., trauma or recent surgery). Staphylococcus aureus is implicated in most patients with acute hematogenous osteomyelitis. Staphylococcus epidermidis, S. aureus, Pseudomonas aeruginosa, Serratia marcescens and Escherichia coli are commonly isolated in patients with chronic osteomyelitis. For optimal results, antibiotic therapy must be started early, with antimicrobial agents administered parenterally for at least four to six weeks. Treatment generally involves evaluation, staging, determination of microbial etiology and susceptibilities, antimicrobial therapy and, if necessary, debridement, dead-space management and stabilization of bone. PMID- 11430457 TI - Benefit of IL-2 therapy in HIV patients confirmed. PMID- 11430458 TI - AHA recommendations for the management of intracranial aneurysms. Agency for Health Care Policy and Research. PMID- 11430459 TI - Antiretroviral approved for use in infants. PMID- 11430460 TI - Synaptic and non-synaptic AMPA receptors permeable to calcium. AB - For a long time, alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA) receptors permeable to calcium have been considered to be either non-existent or as "atypical". There is now ample evidence that these receptors exist in numerous regions of the nervous system and in many neuronal as well as non-neuronal cell populations. This evidence has been accumulated by several methods, including electrophysiological recording, calcium imaging and cobalt-loading. Functional AMPA receptors permeable to calcium are already expressed at very early stages of embryonic development, well before the onset of synaptogenesis. They are probably involved in the paracrine signaling necessary for construction of the nervous system before becoming involved in synaptic transmission. In immature cells, cyclothiazide strongly increases the steady-state level of responses not only to AMPA, but also to kainate. Ingestion, during pregnancy, of food or drug substances that can cross the placental barrier and act upon the embryonic receptors may constitute a risk for normal development. In the adult nervous system, synaptic as well as non-synaptic (paracrine) AMPA receptors permeable to calcium are probably widely expressed in both glial and neuronal cells. They may also participate in controlling some aspects related to adult neurogenesis, in particular the migration of newly formed neurons. PMID- 11430461 TI - Ca2+-images of smooth muscle cells and endothelial cells in one confocal plane in femoral artery segments of the rat. AB - Simultaneous recording of Ca2+-images in one confocal plane from vascular smooth muscle cells (SMCs) and endothelial cells (ECs) of an intact rat femoral artery segment was performed using indo-1 and a confocal microscope. During application of 10 microM acetylcholine (ACh), elevation and oscillation of intracellular Ca2+ concentration ([Ca2+]i) were observed in ECs but not in SMCs. Sequential conduction of Ca2+ oscillation from an EC to the neighboring ECs in one longitudinal direction was often observed in the presence of ACh. On the other hand, the activation of voltage-dependent Ca2+ channels by external 30 mM K+ resulted in the elevation of [Ca2+]i only in SMCs. When 10 microM ACh was added in the presence of 30 mM K+, it was observed in one confocal plane that [Ca2+]i in ECs and SMCs was almost simultaneously increased and decreased, respectively. The simultaneous recording method in this intact preparation will provide a line of valuable information about the interactions between SMCs and ECs, based on spatio-temporal analyses of absolute values of [Ca2+]i in individual cells. PMID- 11430462 TI - Slowing Na+ channel inactivation prolongs QT interval and aggravates adrenaline induced arrhythmias. AB - We investigated the effects of prolonged repolarization induced by slowed inactivation of Na+ channel on adrenaline-induced arrhythmias in halothane anesthetized, closed-chest dogs. We used sea anemone toxins (ATX-II and Anthopleurin-A) to prolong ventricular repolarization and examined their effects on adrenaline arrhythmias. Sea anemone toxins prolonged the QTc- and JTc intervals (P<0.01), but did not affect the PQ interval, QRS duration, heart rate and mean blood pressure. Although sea anemone toxins did not induce any arrhythmias by themselves, under the treatment with these toxins, arrhythmias were induced by non-arrhythmia-inducing doses of adrenaline in four dogs out of seven and the control arrhythmias induced by adrenaline were aggravated. These results indicate that, similar to the inhibition of K+ channels by class III drugs, which we have already reported, slowing Na+ channel inactivation with QTc prolongation also aggravates adrenaline-induced arrhythmias. PMID- 11430463 TI - NTE-122, an acyl-coa:cholesterol acyltransferase inhibitor, prevents the progression of atherogenesis in cholesterol-fed rabbits. AB - The cholesterol-lowering and anti-atherosclerotic effects of NTE-122 (trans-1,4 bis[[1-cyclohexyl-3-(4-dimethylaminophenyl)ureido]methyl]cyclohexane), an acyl CoA:cholesterol acyltransferase (ACAT) inhibitor, were evaluated in 1% cholesterol diet-fed rabbits. NTE-122 (1, 3 and 10 mg/kg per day) lowered the total cholesterol levels in both plasma and liver dose-dependently (by 99% and 94% at 10 mg/kg per day, respectively). In the aortic wall of the rabbits given NTE-122, the atherosclerotic lesion area in both aortic arch and thoracic aorta were dose-dependently reduced (by 100% at 10 mg/kg per day), and the total cholesterol content in aortic arch was also lowered significantly at more than 3 mg/kg per day. These results suggest that NTE-122 is capable of exhibiting anti atherosclerotic effects. PMID- 11430464 TI - An orally active chymase inhibitor, BCEAB, suppresses heart chymase activity in the hamster. AB - We investigated the effects of a novel chymase inhibitor, BCEAB (4-[1-[[bis-(4 methyl-phenyl)-methyl]-carbamoyl]-3-(2-ethoxy-benzyl)-4-oxo-azetidine-2-yloxy] benzoic acid). The IC50 value of BCEAB for purified human chymase was 5.4 nM, whereas BCEAB did not inhibit the angiotensin-converting enzyme, elastase and tryptase. In isolated dog arteries, the IC50 value of BCEAB for the angiotensin I induced contraction in the presence of 1 microM lisinopril was 2.8 microM. In the hamster, the heart chymase activities were significantly suppressed to 42.0% and 26.9% 3 h after oral administration of 100 and 300 mg of BCEAB/kg of body weight, respectively. In conclusion, BCEAB is a useful chymase inhibitor for studying the role of chymase in vivo. PMID- 11430465 TI - Postprandial hyperlipidemia in Zucker diabetic fatty fa/fa rats, an animal model of type II diabetes, and its amelioration by acyl-CoA:cholesterol acyltransferase inhibition. AB - Postprandial hyperlipidemia (PH) is frequently observed in diabetic patients. We performed an oral fat-loading test in Zucker diabetic fatty (ZDF) fa/fa rats, a model for type II diabetes, to determine whether PH was induced in the rats. Post fat-loading changes in serum cholesterol and triglyceride levels were significantly greater in the fa/fa rats than those seen in their lean littermates and an acyl-CoA:cholesterol acyltransferase (ACAT) inhibitor significantly reduced these levels by 24% and 31%, respectively. Therefore, we confirmed that PH appeared in ZDF fa/fa rats by fat loading and ACAT inhibition may be a potential treatment for PH. PMID- 11430466 TI - Suppressive effect of tranilast on interleukin-5 prolonged eosinophils survival via apoptosis. AB - Tranilast has long been used clinically to treat allergic diseases such as bronchial asthma. To further clarify the antiinflammatory machanism, we examined the ability of tranilast to counteract the prolongation of eosinophil survival induced by interleukin (IL)-5. Tranilast reduced the IL-5 prolonged survival of eosinophils at the concentration range of 30 microg/ml to 100 microg/ml. The DNA extracted from eosinophils cultured with tranilast showed signs of fragmentation that was comparable with apoptosis. Electron-microscopic analysis of activated eosinophils cultured with 100 microg/ml of tranilast also revealed morphologic features of apoptosis. These data suggest that tranilast may act in vivo on activated eosinophils to reduce inflammation in allergic diseases. PMID- 11430467 TI - Method for recording spinal reflexes in mice: effects of thyrotropin-releasing hormone, DOI, tolperisone and baclofen on monosynaptic spinal reflex potentials. AB - Mice were used to record the spinal reflex potentials and to examine the effects of some drugs upon them. In anesthetized mice, laminectomy was performed in the lumbo-sacral region, and monosynaptic reflex potential (MSR) and polysynaptic reflex potential were recorded from the L5 ventral root after stimulation of the L5 dorsal root. Thyrotropin-releasing hormone (TRH) and 1-(4-iodo-2,5 dimethoxyphenyl)-2-aminopropane hydrochloride (DOI) produced transient and long lasting increases in the MSR amplitude, respectively. Tolperisone hydrochloride and baclofen produced transient and long-lasting MSR depressions, respectively. These results show that mice can be used to record spinal reflex potentials, and that it may be possible to study the spinal cord function of mutant and knockout mice using this method. PMID- 11430468 TI - Modulation of hippocampal long-term potentiation by the amygdala: a synaptic mechanism linking emotion and memory. AB - Why are emotionally arousing experiences well-remembered? Since the amygdala and hippocampus play pivotal roles in emotion and memory, respectively, the interaction between these brain regions may underlie the formation of enhanced memory for emotionally arousing events. Behavioral experiments using animals have demonstrated that lesions of the amygdaloid nuclei or infusions of drugs into the amygdaloid nuclei impair or enhance hippocampal-dependent learning. In addition, we have obtained direct evidence that neural inputs from the amygdala modulate synaptic plasticity in the hippocampus, through electrophysiological experiments using anesthetized rats. Electrical stimulation of the basolateral amygdala evoked synaptic potentials in the dentate gyrus of the hippocampus, indicating that there is a neural connection from the amygdala to the hippocampus. Lesion of the basolateral or basomedial, but not central, amygdala resulted in attenuation of long-term potentiation (LTP) at the perforant path-dentate gyrus granule cell synapses. High-frequency stimulation of the basolateral or basomedial amygdala alone did not induce LTP in the dentate gyrus, but facilitated the induction of LTP when applied at the same time as tetanic stimulation of the perforant path. The activity-dependent facilitation of hippocampal LTP by the basomedial and basolateral amygdala may be a synaptic mechanism underlying memory enhancement associated with emotions. PMID- 11430469 TI - Electrophysiologic effects of an antiarrhythmic agent, bidisomide, on sodium current in isolated rat ventricular myocytes: comparison with mexiletine and disopyramide. AB - The effects of bidisomide, an antiarrhythmic agent, on sodium current (I(Na)) in isolated rat ventricular myocytes were investigated using a whole cell voltage clamp method. Bidisomide blocked I(Na) with a Ki of 214 microM at a holding potential of -140 mV. The blockade of I(Na) was enhanced at a less negative holding potential of -100 mV with a Ki of 21 microM. Bidisomide shifted the steady state inactivation curve to a negative potential direction by 20 mV without a significant change in the slope factor. Bidisomide slowed the time course of recovery of I(Na) at a holding potential of -140 mV with a slow recovery phase. The time constant of recovery phase for bidisomide, disopyramide and mexiletine were 2703, 1858 and 757 ms, respectively. The development of the block of I(Na) consisted of two phases in the presence of bidisomide. The fast and slow time constants were 11 and 648 ms. Bidisomide produced a use-dependent block of I(Na) when the depolarizing pulse was repeated at 1-3 Hz. Our results indicate that bidisomide binds to rat cardiac sodium channels and that the dissociation kinetics of bidisomide from the inactivated sodium channel is slower than that of disopyramide. PMID- 11430470 TI - Mechanism of atropine-resistant contraction induced by Dai-kenchu-to in guinea pig ileum. AB - To clarify the contractile mechanism of Dai-kenchu-to, the effects of hydroxy beta-sanshool (an ingredient of Zanthoxylum fruit), Zanthoxylum fruit (a constituent herb of Dai-kenchu-to) and Dai-kenchu-to were studied in mucosa-free longitudinal muscle of guinea pig ileum. Hydroxy beta-sanshool at 10(-7)-10(-5) g/ml induced dose-related contractions accompanied by autonomous contraction and produced an initial contraction at a concentration of 10(-4) g/ml or more. The contraction induced by hydroxy beta-sanshool (10(-5) g/ml) was significantly inhibited by tetrodotoxin or the capsaicin-receptor antagonist capsazepine. Although atropine or the substance P antagonist spantide tended to inhibit the contraction, a combination of atropine and spantide almost abolished the contraction by hydroxy beta-sanshool. The P2-purinoceptor antagonist pyridoxal phosphate-6-azophenyl-2',4'-disulphonic acid did not affect hydroxy beta-sanshool induced contraction in the presence or absence of spantide. The tonic contractions by Zanthoxylum fruit (2 x 10(-4) g/ml) and Dai-kenchu-to (10(-3) g/ml) were significantly inhibited or tended to be inhibited by atropine, spantide, tetrodotoxin or capsazepine and were remarkably suppressed by the combination of atropine and spantide. These results suggested that acetylcholine release from intrinsic cholinergic nerves and tachykinins from sensory neurons are involved in the contractions induced by hydroxy beta-sanshool and that tachykinins may be involved in the atropine-resistant contraction by Dai-kenchu to. PMID- 11430471 TI - Do mucosal mast cells contribute to the immediate asthma response? AB - In rat trachea, two types of mast cells have been identified, connective tissue mast cells (CTMCs) and mucosal mast cells (MMCs). We previously reported that CTMCs play an important role in tracheal contraction in vitro via 5 hydroxytryptamine (5-HT) release in a rat model. In this study, we investigated whether MMCs also play a role in tracheal contraction by employing mast cell deficient (Ws/Ws) rats and their congenic (+/+) rats. Rats were actively sensitized with ovalbumin and challenged with it 2 weeks later. To exclude the influence of CTMCs, rats were pretreated for 7 days with compound 48/80 injected i.p. in increasing doses. Histological study confirmed that degranulation occurred in CTMCs, but MMCs still remained. Histamine levels in trachea decreased to 9.31% of control levels. Ovalbumin-specific IgE production showed a time dependent increase in both Ws/Ws and +/+ rats after sensitization with no significantly different values between the two groups. Ovalbumin challenge caused contraction of the trachea in sensitized control (+/+) rats, but not in sensitized Ws/Ws and compound 48/80-pretreated +/+ rats. Ketanserin inhibited the contraction, but leukotriene antagonist ONO-1078 did not, indicating that the contraction was due to 5-HT, whereas leukotriene, a mediator specific derived from MMCs, has no significant effect. The results suggest that MMCs has minimal, if any, contribution to tracheal contraction and might have another function. Furthermore, Ws/Ws and the congenic rats provide a good model for studying the role of mast cells in the immunologic response in airways. PMID- 11430472 TI - Modification of the effects of benzodiazepines on the exploratory behaviors of mice on a hole-board by diabetes. AB - The effect of diabetes on the emotional behavior of mice was examined using an automatic hole-board apparatus. Changes in the emotional state of mice were evaluated in terms of changes in exploratory activity; i.e., total locomotor activity, numbers and duration of rearing and head-dipping, and latency to the first head-dipping. The number and duration of head-dipping in diabetic mice were less than those in non-diabetic mice. Diazepam (0.1-0.56 mg/kg, i.p.) dose dependently increased the number and duration of head-dipping at doses that did not produce sedation in both diabetic and non-diabetic mice. In contrast, methyl beta-carboline-3-carboxylate (1 and 2 mg/kg, i.p.) decreased the number and duration of head-dipping in non-diabetic mice, but not in diabetic mice. The number and duration of head-dipping in diabetic mice were increased by treatment with flumazenil (0.1 and 0.3 mg/kg, i.v.). These doses of flumazenil did not affect the number or duration of head-dipping in non-diabetic mice. The present data indicate that diabetic mice exhibited anxiety in the hole-board test and that a benzodiazepine receptor antagonist affected the attenuated number and duration of head-dipping in diabetic mice. The heightened anxiety in diabetic mice may be due to the dysfunction of the benzodiazepine receptor and/or of central inhibitory systems. PMID- 11430473 TI - VUF-K-8788, a periphery-selective histamine H1 antagonist with anti-pruritic activities. AB - The pharmacological properties of 7-[3-[4-(2-quinolinylmethyl)-1-piperazinyl] propoxy]-2,3-dihydro-4H-1,4-benzothiazin-3-one (VUF-K-8788) were investigated in vitro and in vivo. VUF-K-8788 inhibited [3H]-mepyramine from binding to the cell membrane of lung parenchyma (Ki value: 5.0 nM) and the histamine-induced contraction of isolated guinea pig ileum (pA2: 9.71) without affecting ileal contractions induced by acetylcholine, serotonin, KCl and BaCl2. The increase of vascular permeabilities induced by histamine and passive cutaneous anaphylaxis (PCA) in guinea pigs were inhibited by VUF-K-8788 in a dose-dependent fashion (ED50: 0.24 and 0.26 mg/kg, p.o., respectively). Moreover, the anti-histaminic effect of VUF-K-8788 was also observed in rats. In experiments on the effects on the central nervous system, VUF-K-8788 at 1 mg/kg, p.o. hardly antagonized the H1 receptor at all in the cerebral cortex of guinea pigs. VUF-K-8788 inhibited the PCA-induced scratching behavior completely without affecting thiopental-induced sleep in mice. These results suggested that VUF-K-8788 would be useful in the treatment of allergic disorders such as atopic dermatitis and eczema. PMID- 11430474 TI - Localization of metallothionein (MT) and expression of MT isoforms induced by cadmium in rat dental pulp. AB - We investigated the induction of metallothionein (MT) by cadmium (Cd) in the dental pulp of rat incisors. Time-course studies of MT mRNA expression after single Cd injection were observed by Northern-blot analysis. The isoform-specific expressions of MT mRNAs (MT-I, MT-II and MT-III) were observed using the reverse transcriptase-polymerase chain reaction (RT-PCR) method. Both MT-I and MT-II mRNA levels increased within 3 h, peaked at 3 h and then decreased. These findings demonstrated that MT-I and MT-II mRNA were rapidly induced by Cd in dental pulp. MT-III mRNA was constitutively expressed in rat dental pulp, but the expression level did not change by Cd treatment. The localization of MT protein in Cd treated rat dental pulp was determined by immunohistochemical staining using anti MT antibody against MT-I and MT-II. MT protein was localized in the specific cell type of odontoblasts (secretory odontoblasts and resting odontoblasts). In conclusion, it is likely that stained MT in the immunohistochemical study should be MT-I and/or MT-II. Furthermore, MT-I and/or MT-II in Cd-treated rat dental pulp was localized in odontoblasts, in which accumulation of Cd were reported. The cell-specific synthesis of MT may be associated with its metal storage and detoxification role in dental tissues. PMID- 11430475 TI - In vivo assessment of the regulatory mechanism of cholinergic neuronal activity associated with motility in dog small intestine. AB - Intestinal motor activity associated with acetylcholine (ACh) release was assessed in the small intestine of anesthetized dogs by simultaneous measurement of motor activity and local ACh concentrations within the intestinal wall with in vivo microdialysis. Basal concentration of ACh measured in the dialysate was 1.12 +/- 0.08 pmol/15 min (n = 10), a value that remained constant until 3 h after perfusion. Intra-arterial infusion of tetrodotoxin reduced dialysate ACh concentration, while the motor activity accelerated at the early phase after infusion of tetrodotoxin and then decreased, thereby suggesting that the motor activity is regulated by not only excitatory cholinergic neurons, but also inhibitory neurons. Intraarterial infusion of atropine increased dialysate ACh concentration but reduced motor activity, thereby indicating that the cholinergic neurons are tonically active and the muscarinic autoreceptors operate to inhibit the ACh release. Intraarterial infusion of norepinephrine reduced, but yohimbine increased both motor activity and dialysate ACh concentration, thereby indicating that the adrenergic neurons regulate the motor activity due to control of cholinergic neuronal activity. This in vivo microdialysis method demonstrated in the whole body of animals that the activity of cholinergic neurons was physiologically regulated by itself and adrenergic neurons. PMID- 11430476 TI - Comparison of the angiotensin II type 1-receptor antagonist YM358 and the angiotensin-converting enzyme inhibitor enalapril in rats with cardiac volume overload. AB - We evaluated the effects of chronic oral administration of an angiotensin II type 1 (AT1)-receptor antagonist YM358 and an angiotensin converting enzyme inhibitor enalapril on hemodynamics and cardiac hypertrophy in rats with volume overload induced heart failure. We assessed changes of cardiac hemodynamics and cardiac hypertrophy at 2 and 4 weeks after administration of YM358 (3, 30 mg/kg per day) or enalapril (30 mg/kg per day) in abdominal aortocaval shunt rats. YM358 (30 mg/kg) attenuated increases of left ventricle (LV)/body weight (BW), left atrium (LA)/BW, right ventricle (RV)/BW and heart/BW ratios, but did not affect cardiac hemodynamics in shunt rats. Enalapril also reduced the increased LV/BW and heart/BW ratios together with significant reductions of systolic blood pressure, left ventricular systolic pressure and the first derivative of left ventricular pressure. These data suggest that the effects on attenuation of the development of cardiac hypertrophy are not different for YM358 and enalapril, although the effects on cardiac hemodynamics are different for the same dosages. The attenuating action of YM358 on cardiac hypertrophy was independent of the action on hemodynamics and indicated the direct action of the AT1 receptor on the heart. PMID- 11430477 TI - Incadronate and etidronate accelerate phosphate-primed mineralization of MC4 cells via ERK1/2-Cbfa1 signaling pathway in a Ras-independent manner: further involvement of mevalonate-pathway blockade for incadronate. AB - Two types of bisphosphonates (BPs), incadronate (INC) and etidronate (ETI) accelerated phosphate (Pi)-primed mineralization of MC4 cells in a subnanomolar dose range. Intracellular signaling pathways involved were examined. 1) The effect of INC but not ETI was partially suppressed by two mevalonate (MVA) pathway metabolites, farnesylpyrophosphate (FPP) and geranylgeranylpyrophosphate (GGPP). 2) The BP-like accelerating effect was produced by statins and also by Toxin B, a Rho GTPases-specific inhibitor. 3) INC induced Cbfa1-nuclear localization within hours; and in an in vivo experiment using ovariectomized mice, its 3 weeks dosing exhibited the same effect in tibial extracts. 4) BPs promoted luciferase expression in murine p1.3-osteocalcin gene 2-luc and p6 osteoblast specific element 2-luc transfected cells, just as MVA, FPP and GGPP did independently and additively to INC. 5) BPs activated extracellular signal regulated kinase (ERK1/2) in a Ras-independent manner within 5 min, and Pi was found to sensitize MC4 cells to BPs. MVA and its metabolites also activated ERKs but in a Ras-dependent manner and additively to INC. Ras dependency was determined using N17Ras-transfected cells. A MEK (MAP kinase-ERK kinase)-specific inhibitor PD98059 alone partly and with FPP completely blocked INC-induced mineralization. The results suggest that BPs act on Pi-sensitized MC4 cells to accelerate mineralization via nonRas-MEK-ERK1/2-Cbfa1 transactivation pathway and INC additionally acts by inhibiting the MVA pathway. PMID- 11430478 TI - Roles of mast cells and histamine in mosquito bite-induced allergic itch associated responses in mice. AB - We investigated itch-associated responses (scratching) to mosquito bites and the role of histamine and mast cells in mosquito-induced itching in mice. Although the first bites of mosquito Aedes albopictus did not increase scratching, repeated bites increased scratching. The response was not diminished even after an interval of 2 months. Similarly, repeated intradermal (i.d.) injections of salivary gland extract (SGE) from Aedes albopictus increased scratching after SGE injection itself and mosquito bites. The scratching peaked within 10 min and almost subsided by 60 min. The opioid antagonist naloxone (1 mg/kg, s.c.) inhibited scratching following SGE injection. Although the non-sedative H1 histamine-receptor antagonist terfenadine (30 mg/kg, p.o.) significantly suppressed scratching induced by histamine (100 nmol/site, i.d.) in either naive or mosquito-sensitized mice, it did not affect mosquito-induced scratching in mosquito-sensitized mice. Repeated injections of SGE increased scratching in mast cell-deficient (WBB6F1-W/Wv) mice as well as in normal (WBB6F1-+/+) littermates. Repeated exposure to mosquito bites roughly doubled serum concentrations of total IgE and IgG1, but not IgG2a. Repeated injections of SGE markedly increased plasma extravasation induced by mosquito bites and such an increase was almost completely suppressed by terfenadine (30 mg/kg, p.o.). The results show the presence of histamine-mediated and histamine-independent mechanisms in cutaneous itching and suggest that histamine probably released from mast cells does not play an important role in itching in immediate allergic reaction. Our murine model of mosquito itching may be useful for studying the mechanisms of immediate allergic itching. PMID- 11430479 TI - Fluctuating asymmetry for specific bristle characters in notch mutants of Drosophila melanogaster. AB - Asymmetry has been used as a measure of developmental stability for bilaterally symmetrical organisms. Most studies have failed to partition the genetic and environmental contributions to the asymmetry phenotype due to the limitations of the systems used or the shortcomings in experimental design. The Notch mutants of Drosophila melanogaster were used to study the genetic contribution to asymmetry for six different bristle characters. Asymmetry response was character specific for the mutants examined. For Nspl, N(Co), N264-47, Ax71d, Ax9B2, AxE2, 1(1)N(B) and nd2 significant asymmetry responses, relative to wildtype Canton-S, were observed for some characters. N60g11 and nd1 did not exhibit significant asymmetry for any of the characters examined. All of the mutants except N60g11 and nd1 showed thoracic bristle asymmetry. However, when asymmetry scores were pooled over the five bristle characters which individually exhibited fluctuating asymmetry, no significant differences were found between any genotypes. Therefore pooling asymmetry values across characters obscures the significant character specific asymmetry values observed. Thus caution is necessary when using the asymmetry phenotype of specific characters to draw organism wide conclusions about developmental stability. PMID- 11430480 TI - Accentuated polymorphism of heterochromatin and nucleolar organizer regions in Astyanax scabripinnis (Pisces, Characidae): tools for understanding karyotypic evolution. AB - Astyanax scabripinnis has been considered a species complex because it presents high karyotypic and morphological variability among its populations. In this work, individuals of two A. scabripinnis populations from different streams in the same hydrographic basin were analyzed through C-banding and AgNOR. Although they present distinct diploid numbers, they show meta and submetacentric chromosome groups highly conserved (numerically and morphologically). Other chromosomal characteristics are also shared by both populations, as the pattern of constitutive heterochromatin distribution (large blocks in the telomeric regions of subtelocentric and acrocentric chromosomes) and some nucleolar chromosomes. Inter-individual variations both in the number and size of heterochromatic blocks, and in the number and localization of NORs were verified in the studied populations, characterizing them as polymorphics for these regions. The mechanisms involved in the dispersion of heterochromatin and NORs through the karyotypes, as well as the possible events related to the generation of polymorphism of those regions are discussed. Furthermore, relationships between these populations and within the context of the scabripinnis complex are also approached. PMID- 11430481 TI - Spatial differences in patterns of modification: selection on hairy in Drosophila melanogaster wings. AB - Artificial selection was carried out for over 45 generations to enhance and suppress expression of the mutation hairy on the Drosophila melanogaster wing. Whole chromosome mapping of X-linked and autosomal modifiers of sense organ number displayed regional differences in magnitude and direction of their effects. Regional specificity of modifier effects was also seen in some interchromosomal interactions. Scanning electron microscopy allowed precise measurement of sense organ size and position along the L3 longitudinal wing vein. Sense organ size varied in a predictable fashion along the proximal-distal axis, and the dorsal pattern differed from the ventral pattern. The high and low selection lines differed most in the proximal portion of the L3 vein. Extra sense organs in the High line were often associated with vein fragments at locations predicted from ancestral vein patterns. Thus, regional specificity of polygenic or quantitative trait locus modifier effects was identified in several different parts of the wing. PMID- 11430482 TI - Strategies for genetic mapping of categorical traits. AB - The search for efficient and powerful statistical methods and optimal mapping strategies for categorical traits under various experimental designs continues to be one of the main tasks in genetic mapping studies. Methodologies for genetic mapping of categorical traits can generally be classified into two groups, linear and non-linear models. We develop a method based on a threshold model, termed mixture threshold model to handle ordinal (or binary) data from multiple families. Monte Carlo simulations are done to compare its statistical efficiencies and properties of the proposed non-linear model with a linear model for genetic mapping of categorical traits using multiple families. The mixture threshold model has notably higher statistical power than linear models. There may be an optimal sampling strategy (family size vs number of families) in which genetic mapping reaches its maximal power and minimal estimation errors. A single large-sibship family does not necessarily produce the maximal power for detection of quantitative trait loci (QTL) due to genetic sampling of QTL alleles. The QTL allelic model has a marked impact on efficiency of genetic mapping of categorical traits in terms of statistical power and QTL parameter estimation. Compared with a fixed number of QTL alleles (two or four), the model with an infinite number of QTL alleles and normally distributed allelic effects results in loss of statistical power. The results imply that inbred designs (e.g. F2 or four-way crosses) with a few QTL alleles segregating or reducing number of QTL alleles (e.g. by selection) in outbred populations are desirable in genetic mapping of categorical traits using data from multiple families. PMID- 11430483 TI - High-density molecular map of chromosome region harboring stripe-rust resistance genes YrH52 and Yr15 derived from wild emmer wheat, Triticum dicoccoides. AB - Two stripe-rust resistance genes, YrH52 and Yr15, derived from the Israeli wild emmer wheat, Triticum dicoccoides, have been located on chromosome 1B. The main objectives of the present study were to increase marker density in the vicinity of YrH52 gene by means of AFLP, RAPD and microsatellite markers, to improve the map of another T dicoccoides-derived stripe-rust resistance gene Yr15 using microsatellite markers, and to preliminarily discriminate these two genes. Additional 26 marker loci comprising 20 AFLPs, three RAPDs, and three microsatellites were found to be linked to YrH52 gene. An updated genetic map consisting of 45 marker loci, in the region of YrH52 gene, was constructed with a total map length of 107.7 cm. The mean interval length was 0.96 cm in the region Xgwm359b-P55M53b carrying YrH52 gene. YrH52 was bracketed by Xgwm413 (Nor1 and UBC212a) and Xgwm273a (Xgwm273d) with map distance of 1.3 and 2.7 cm from either side, respectively. Eight additional microsatellite markers were found to be linked with Yr15, and the linkage map of Yr15 gene was thus obviously improved. In the YrH52-mapping population, no crossover was detected in the interval UBC212a (Xgwm413)-Yr15-Nor1, and YrH52 was located distally outside this interval. It may suggest that YrH52 is different from Yr15 even though both of them are derived from T. dicoccoides and are mapped on chromosome 1BS. The large number of molecular makers revealed in the present study would be helpful for the marker-assisted introgression of the T. dicoccoides-derived YrH52 and Yr15 stripe rust resistance genes into elite cultivars of wheat, and the high-density map would accelerate the map-based cloning of the two genes. PMID- 11430484 TI - Chromosomal responses of blowfly Lucilia cuprina to heat and heavy metal stress. AB - Chromosomal responses to heat and heavy metal shocks were studied in the pupal trichogen polytene chromosomes of Lucilia cuprina. Heat shock induced seven distinct puffs on different chromosomes at the following loci: 1C2, 21A1, 21C, 23B, 24B, 42A and 95B3. Arsenate and mercury, two of the most common toxic environmental chemical pollutants also, induced almost the same set of puffs except 20B2, which appeared to be induced by metals only and 95B3, which was not induced by arsenate. The findings suggest that a common set of gene loci encoding the heat shock proteins is responsive to these diverse environmental stresses. PMID- 11430485 TI - Karyotypic analysis of the cotton boll weevil, Anthonomus grandis Boheman. AB - The diploid chromosome number of the cotton boll weevil, Anthonomus grandis Boheman, is 44. Both C- and N-banding techniques of mitotic cells demonstrated constitutive heterochromatin in the p arm of the eight largest chromosomes, the p arm of the X chromosome, and the centromeric region of autosomal groups A-D. Neither the y nor the group E autosomes appeared to contain constitutive heterochromatin. Supernumerary chromosomes were not found in the boll weevil. Restriction endonuclease banding of primary spermatocytes revealed a rod-shaped Xy tetrad in which the X and y were terminally associated. The p arm of the large, submetacentric X was C-band positive. While two of the autosomal tetrads were typically ring-shaped in primary spermatocytes, the remaining 19 autosomal tetrads were rod-shaped. PMID- 11430486 TI - Random amplified polymorphic DNA (RAPD) analysis in Indian mung bean (Vigna radiata (L.) Wilczek) cultivars. AB - Greengram [Vigna radiata (L.) Wilczek], also known as mung bean, widely cultivated in a large number of countries, is an important pulse crop of Asia and is considered one of the ancestral species of the genus Vigna. Since yields of greengram have remained low across subtropical and tropical Asia, it is important to estimate genetic diversity in existing cultivars in order to see if the lack of genetic variability might be a constraining factor. In this study, 32 Indian cultivars of greengram were subjected to random amplified polymorphic DNA (RAPD) analysis using 21 decamer primers. A total of 267 amplification products were formed at an average of 12.71 per primer with an overall polymorphism of 64%. The extent of polymorphism was moderate to low. Jaccard similarity coefficient values ranged from 0.65 to 0.92. The cluster analysis resulted in mainly three clusters revealing greater homology between cultivars released from the same source. The results of principal components analysis also substantiated this conclusion. The close genetic similarity between the cultivars could be explained due to the high degree of commonness in their pedigrees. The narrow genetic base of the greengram cultivars revealed in the present analysis emphasises the need to exploit the large germplasm collections having diverse morphoagronomic traits in cultivar improvement programs. PMID- 11430487 TI - Genetic and biochemical analysis of brown eye mutation in Drosophila nasuta nasuta and Drosophila nasuta albomicans. AB - By analyzing the progeny of crosses involving brown eye mutants and the wild types in two members of Drosophila nasuta subgroup namely D. n. nasuta and D. n. albomicans we could show that the mutant gene is recessive, located in the chromosome 2 and the alleles of this gene are present at different loci. A study of fitness in the eye color mutants in comparison with the wild types revealed that D. n. nasuta mutant has higher viability at both 25+/-1 degrees C and ambient temperatures; while D. n. albomicans mutant has faster rate of development only at 25+/-1 degrees C. Quantitative analysis of eye pigments in the mutants revealed that there is biosynthesis of both pteridines and xanthommatins unlike in bw/bw of D. melanogaster, where only xanthommatins are synthesized. In both the species, the pteridine quantities in mutants are similar; whereas xanthommatin quantity in bw(n)/bw(n) is 10 times higher than that of bw(a)/bw(a). Further, the F1 progeny of intraspecific crosses (wild type X mutant) are found to have high amounts of pteridine, even when compared with parental wild type. PMID- 11430488 TI - Genetic analysis of extended lifespan in Drosophila melanogaster III. On the relationship between artificially selected and wild stocks. AB - Adult lifespans, age-specific survival, age-specific mortality, survival times on paraquat, and survival times on DDT were assayed in seven lines of Drosophila melanogaster, including two genetically heterogeneous wild lines recently collected from nature, and three inbred and recombinant inbred lines derived from an artificial selection experiment for increased lifespan. Survival on paraquat is positively correlated with adult lifespan. DDT resistance is uncorrelated with either paraquat resistance or lifespan. The wild lines are unexceptional with respect to average lifespan, paraquat resistance, age-specific survivorship, and leveling off of mortality rates at advanced ages, but have high levels of DDT resistance. Cluster analysis groups the wild lines with three unselected laboratory stocks in one cluster, while two long-lived elite recombinant inbred lines form a second cluster. Long-lived laboratory-adapted lines are quantitatively differentiated from the wild stocks, both with respect to average adult lifespans and resistance to an oxidizing agent. We reject the 'recovery' hypothesis, which proposes that Drosophila artificially selected for long life have phenotypes that merely recover the wild state. PMID- 11430489 TI - Unexpected low genetic differentiation among allopatric species of section Algarobia of Prosopis (leguminosae). AB - The Chaquena Biogeographic Province, in South America, is the main diversity centre of Prosopis. A group of sympatric species of Section Algarobia in this region constitutes a syngameon, characterised by frequent hybridization and introgression. These processes have been postulated as responsible for the low genetic differentiation observed among species within this group. In this study genetic variability and differentiation among geographically isolated species of the same section was analysed through isoenzyme electrophoresis. Variability parameters and fixation indices were estimated to determine the genetic structure of populations. Two Argentinean, P. ruscifolia ('vinal') and P. flexuosa ('algarrobo amarillo'), and one North American species, P. glandulosa (honey mesquite) were studied. All of them showed, similarly to other species of the same section, high genetic variability and exhibit homozygote excess, probably due to population substructure and low rates of selfing. In contrast to our hypothesis, genetic similarity among species is not related to geographic distance. Genetic distances between P. glandulosa and South American species are similar to those observed among species of this subcontinent. The results obtained suggest that the high genetic similarity among the species of the section Algarobia studied is not due to hybridization. PMID- 11430490 TI - Cytogenetic studies on Apareiodon affinis (Pisces, Characiformes) from Parana river basin: sex chromosomes and polymorphism. AB - Comparative cytogenetic studies were carried out on Apareiodon affinis from an important hydrographic system at South America, the Parana river basin. Two distant regions were chosen, which were separated by Guaira Falls (formerly Sete Quedas); the region in the upper part of the hydrographic basin is called Upper Parana (Brazil), whereas and the other in the lower part is called Lower Parana (Argentina). Individuals from Upper Parana have diploid numbers of 2n = 54 (NF= 108) for males and 2n = 55 (NF = 110) for females, showing female heterogamety with a ZZ/ZW1W2 multiple sex chromosomes system that is endemic for the region. In different localities at Lower Parana, the specimens presented diploid number of 2n = 54 for both sexes, without any sex chromosomes heteromorphism. However, they have an accentuated polymorphism characterized by variation in number of acrocentric chromosomes, constituting something new for family Parodontidae. The most likely hypothesis to explain the origin of such polymorphism is based on successive pericentric inversions giving rise to acrocentric chromosomes. Thus, it was possible to detect 10 cytotypes along the Lower Parana basin. Such chromosomal variations possibly are the consequence of an adaptative process. Our data probably indicate the occurrence of distinct species in each region that share the same denomination. PMID- 11430491 TI - Dnop56, a Drosophila gene homologous to the yeast nucleolar NOP56 gene. AB - Small nucleolar RNAs (snoRNAs) are trans-acting factors involved in maturation of rRNA and have been classified into Box C/D and Box H/ACA families. Most of the snoRNAs occur as ribonucleoprotein complexes with snoRNA-associated proteins (snoRNPs). All Box C/D snoRNAs in yeast form complexes with Nop1p, Nop56p and Nop58p. Similarly, it has been reported that Box H/ACA-containing snoRNAs form complexes with yeast Gar1p. Nop56p and Nop58p homologs have been described in several species. Here we report the isolation and molecular characterization of the Dnop56 genes from D. melanogaster and D. subobscura which show a very similar structure. Drosophila Nop56p proteins contain lysine-rich regions at their carboxy-terminus, and show a high degree of similarity to other Nop56p proteins from different organisms. Phylogenetic relationships among these proteins and other snoRNPs have been established. PMID- 11430492 TI - Cavernous sinus thrombophlebitis caused by sphenoid sinusitis--report of autopsy case. AB - A 34-year-old man developed fever, headache, nausea, double vision, exophthalmus, ptosis, disturbance of vision and oculomotor nerve palsy. Magnetic resonance imaging and cerebral angiography led to the clinical diagnoses of cavernous sinus thrombophlebitis and suspicion of bacterial aneurysm of the left internal carotid artery, respectively. Peptostreptococcus was detected in blood culture analysis. He died 15 days after admission. Systemic organs showed several septic changes. In particular, the bilateral cavernous sinuses were enlarged and showed severe neutrophilic leukocyte infiltration of the walls and organization, recanalization and abscesses in thrombi. In anterior to the middle cranial fossa, abscess forming, necrotic, hemorrhagic meningitis, purulent sphenoid sinusitis, pyogenic osteomyelitis of the sphenoid bone, suppurative encephalitis, and inflammatory necrosis of the hypophysis were seen. Based on these findings, we diagnosed the patient with cavernous sinus thrombophlebitis caused by sphenoid sinusitis. PMID- 11430493 TI - TUNEL-positive staining in white and grey matter after fatal head injury in man. AB - Paraffin sections from the hippocampus, the cingulate gyrus and the insula of 18 head-injured patients who survived between 5 hours and 10 days, and 18 age matched controls, were stained by the terminal deoxynucleotidyl transferase mediated biotinylated deoxyuridine triphosphate nick end labelling (TUNEL) technique for evidence of in situ DNA fragmentation. Additional staining techniques (HE, combined LFB/CV and immunohistochemistry for GFAP and CD68) were used to characterize any lesions and their time course. Only the occasional TUNEL+ cell per area was seen in the control brains. TUNEL+ cells were identified in both grey and white matter of the head-injured material and their numbers peaked between 24 and 48 hours and were still present at 10 days. Within the hippocampus, fewer TUNEL+ cells were seen in grey (between 3-5 per area) than in the white matter, (up to 51+ per area) whereas in the cingulate gyrus and in the insula, the number of TUNEL+ cells was always greater in the cortex (between 11 20 per area) than in white matter (6-10 per area). In the grey matter, most TUNEL+ cells had the morphology of necrosis. However, the histological appearances of some of the neurons (2-3%), and of oligodendroglia and macrophages in white matter (about 5%) were those of apoptosis. PMID- 11430494 TI - Tanycytic ependymoma in association with neurofibromatosis type 2. AB - A rare case of tanycytic ependymoma associated with neurofibromatosis type 2 (NF2) is presented for the first time, with emphasis on its clinical course and histopathological features. A 30-year-old man had developed gait disturbance in his childhood, and harbored multiple tumors in spinal nerve roots, in the intradural extramedullary and intramedullary spinal cord. The spinal root tumor and intradural extramedullary tumor were histologically diagnosed as schwannoma and meningioma, respectively. Magnetic resonance imaging showed two intramedullary cystic lesions, one in the cervical and the other in the thoracic spine. Because his sensorimotor dysfunction in the lower extremities continued to worsen gradually, three of the multiple nodular tumors in the thoracic cystic lesion were removed. All three tumors were composed of eosinophilic piloid cells with modest nuclear pleomorphism. No Rosenthal fibers were found. A concentration of slender eosinophilic cellular processes surrounding the vascular wall was seen. Periodic acid Schiff and Masson trichrome-positive balloons were seen in the extracellular space. Detection of ependymal rosettes, although only few in number, led the diagnosis as a tanycytic ependymoma. Recognition of this ependymoma variant should be emphasized to avoid confusion with pilocytic astrocytoma or intramedullary schwannoma. PMID- 11430495 TI - Possible health impact of phytoestrogens and xenoestrogens in food. AB - Plants produce estrogen-like substances, denominated phytoestrogens, which are present in many human foodstuffs. The consumption of phytoestrogens has been associated with a variety of protective effects. Their relative estrogenic potency combined with their concentrations in food and human plasma indicate biological relevance. However, their biological properties differ from those of estradiol or other endogenous estrogens in humans. For instance, their possible effects on SHBG, inhibition of steroid metabolizing enzymes, anti-proliferative and anti-angiogenetic and other side effects have been described. Furthermore, phytoestrogens can exert estrogenic and antiestrogenic activities at the same time and their potency and metabolism have not been yet elucidated in all cases. In recent decades growing evidence has accumulated on the hormone-like effects of synthetic chemicals that appeared in the environment. The possible impact of xenoestrogens, to which humans are also exposed through the food chain, needs to be further clarified as well. The molecular effects and control mechanisms of these substances, their pharmacokinetics, threshold levels and dose-response differences are issues that require further research before a full assessment of their effect on humans can be drawn. Evaluating the total exposure and impact of this estrogenic effect is very challenging because of the lack of specific knowledge in some areas and the differences in the biological activity among these substances, as pinpointed in this review. PMID- 11430496 TI - Human exposure to endocrine disrupters: standardisation of a marker of estrogenic exposure in adipose tissue. AB - In many epidemiological studies based on the direct measurement of exposure to organochlorines, the chemicals of concern are determined directly from adipose tissue samples. Although the measurement of all possible organochlorines, their metabolites, isomers and congeners may be desirable, it is expensive and time consuming and many chemicals with hormonal activity may not yet have been identified. Testing systems are therefore required to screen for estrogenicity and to identify appropriate biomarkers of human exposure. To address this issue, we developed and standardised a method to assess the total estrogenic xenobiotic burden in human adipose tissue. The method extracts and separates the more lipophilic xenoestrogens from ovarian estrogens, with a subsequent bioassay determination of the cumulative effect of the xenoestrogens. It was applied to 400 women, using 200 mg of adipose tissue: 65% of samples showed measurable estrogenicity in the fraction where most non-polar xenoestrogens eluted, and 76% of fractions where ovarian estrogens eluted were positive for estrogenicity. Residues of 16 organochlorine pesticides were determined. No correlation was found between pesticide content and estrogenicity of the samples. The high percentage of positive samples suggests that the method is sensitive enough to be used as a biomarker of human exposure to estrogenic xenobiotics and can be applied in epidemiological studies. PMID- 11430497 TI - Threshold analysis of selected dose-response data for endocrine active chemicals. AB - Using a biologically relevant mathematical model, the Michaelis-Menten equation, we examined published data from endocrine active chemicals for evidence of no threshold dose-response curves. Data were fit to a modified Michaelis-Menten equation which accounted for total background response. Subsequently, the data sets were analyzed using non-linear regression in order to estimate the four parameters of interest (non-hormone controlled background (Bnh), maximum response (Rmax), endogenous hormone level (D0), and the dose at which a half-maximal response was observed (ED50)) and to determine the fit to the fully modified Michaelis-Menten equation. Subsequently, response data were adjusted to account for Bnh and then normalized to Rmax, while dose data were adjusted to account for D0 and then normalized to the ED50. This data set was combined into a single, composite data set and fit to the fully modified Michaelis-Menten equation. We examined 31 data sets (24 endpoints) from studies on 9 different chemical/hormone treatments. Twenty-six of the data sets fit the modified Michaelis-Menten equation with high multiple correlation coefficients (r>0.90). The normalized data demonstrated a good fit to the modified Michaelis-Menten equation. These results indicate that a variety of biological responses fit the modified Michaelis-Menten equation, which does not have a threshold dose term. PMID- 11430498 TI - Identification of enterococcal isolates by temperature gradient gel electrophoresis and partial sequence analysis of PCR-amplified 16S rDNA variable V6 regions. AB - Based on partial sequence analysis of PCR-amplified 16S rDNA variable V6 regions of 14 enterococcal type strains, Enterococcus faecalis, Enterococcus mundtii, Enterococcus gallinarum, Enterococcus avium, Enterococcus raffinosus and Enterococcus saccharolyticus showed characteristic sequence motifs which made it possible to separate them into six individual species lines. Furthermore, two species cluster groups could be identified, including (i) Enterococcus faecium, Enterococcus durans, Enterococcus hirae, Enterococcus malodoratus, and (ii) Enterococcus casseliflavus/Enterococcus flavescens, Enterococcus pseudoavium, Enterococcus dispar and Enterococcus sulfureus. There were identical DNA sequences in the V6 region within each group. Temporal temperature gradient gel electrophoresis (TTGE) of the PCR products from 16 type strains, 12 enterococcal reference strains and 8 clinical isolates revealed that a single nucleotide divergence in DNA sequences was sufficient for separation, identification and division of the studied enterococcal strains into corresponding TTGE profiles. It was concluded that partial DNA sequence analysis and TTGE profiling of PCR amplified 16S rDNA variable V6 regions provide useful tools for the identification of clinically important Enterococcus spp. PMID- 11430499 TI - Performance characteristics and clinical utility of a hybrid ELISA for detection of ANA. AB - To investigate the clinical utility of a newly developed hybrid ELISA for antinuclear antibodies (ANA), a cross-sectional study of patients admitted to the Section of Rheumatology was initiated. The ELISA was compared to indirect immunofluorescence (IIF) on HEp-2 cells. Accuracy of tests was analyzed using receiver-operating characteristic methodology (ROC). In addition, diagnostic sensitivity, specificity and predictive values were calculated for each assay. Results from the ROC analysis showed a slightly superior accuracy for IIF as compared to ELISA. Furthermore, IIF showed higher diagnostic sensitivity and positive predictive value for all combinations of patients and reference populations. This was due to enhanced detection by IIF, in contrast to ELISA, of diagnostically useful antibodies. IIF detected 87.4% and ELISA detected 84.2% of sera with antibodies against extractable nuclear antigens (ENA). In addition, IIF detected diagnostically important antibodies that are not included among the anti ENA. The hybrid ELISA either lacks or does not contain the relevant antigens in sufficient amount. Inclusion of these antigens may further enhance the performance characteristics of the ELISA. PMID- 11430500 TI - Stereological study on the submandibular gland in hypothyroid rats. AB - Most research done on hypothyroidism has focused on physiological and biochemical aspects of the tissues, whilst there has been little work on tissue morphology especially on salivary glands. The present study has used hypothyroid Wistar rats as a model for investigating the effects of hypothyroidism on submandibular gland structure. Two groups of Wistar rats were studied. One was made hypothyroid with methimazole and the second was an untreated euthyroid group (control). They were euthanised after 10 weeks. Submandibular glands were removed and studied. Systematic random transverse sections were obtained from submandibular glands and subjected to morphometric analysis. Volume density and absolute volume of granular, striated and excretory ducts and interlobular connective tissue were estimated by point counting. Volume-weighted mean particle volume of serous and mucous acini was also determined by unbiased stereology. Absolute volume of granular ducts in the submandibular gland of hypothyroid rats was reduced by approximately 50% (p<0.009) whilst that of the striated and excretory ducts and interlobular connective tissue was unaffected. Volume of serous acini was also significantly (p<0.03) lower in hypothyroid rats. These changes suggest that hypothyroidism has an effect on submandibular gland structure, and that this effect occurs mostly in two major exocrine compartments (granular duct and serous acinus) of the gland. PMID- 11430501 TI - Antimicrobial resistance of Shigella sonnei in Korea during the last two decades. AB - Eighty-eight strains of Shigella sonnei isolated in Korea during the period 1980 to 1999 were tested for susceptibility to 13 antimicrobial agents. S. sonnei isolates demonstrated high frequencies of resistance to sulfamethoxazole (97.7%), tetracycline (96.6%), and trimethoprim (95.5%). S. sonnei isolates from the 1990s were more resistant to nalidixic acid than isolates from the 1980s (100 vs 7.7%), while isolates from the 1990s were more susceptible to chloramphenicol than isolates from the 1980s (0 vs 100%). Ampicillin-resistant S. sonnei isolates produced the TEM-1 beta-lactamase with a pI of 5.4. The TEM-1 gene was located on conjugally transferable plasmids in the majority of isolates. S. sonnei isolates were all susceptible to cefotaxime, cefoxitin, ceftazidime, ciprofloxacin, and norfloxacin. These results indicate that cephalosporins and quinolones may be alternative antibiotics for the treatment of S. sonnei infections in Korea. PMID- 11430502 TI - Inverse relation between mRNA synthesis and secretion of parathyroid hormone in athymic mice grafted with human parathyroid tissue. AB - Parathyroid hormone (PTH) mRNA in original and transplanted human adenomatous parathyroid tissue and human serum intact PTH (S-iPTH) was measured in athymic mice at 4, 7, 14, and 28 days after transplantation. Parathyroid tissue was obtained during surgery for hyperparathyroidism and implanted subcutaneously. PTH mRNA detection was done with RT-PCR followed by membrane blot and hybridisation and S-iPTH was analysed using a human specific immunoradiometric method. At 4 days, PTH mRNA was 79.6+/-5.3% (mean+/-SE) of that in original tissue whereas S iPTH was only 5.4 ng/l. At 28 days, PTH mRNA was significantly reduced to 60.7+/ 4.1% whereas S-iPTH was increased to 192 ng/l. The reduced PTH mRNA expression in the transplants at 28 days may be explained by an inhibited DNA transcription. The presence of human S-iPTH in transplanted mice at 4 days may be due to cell disintegration and diffusion. The gradual increase in S-iPTH during the experimental period probably reflects increased transplant cell volume and improved graft revascularisation. PMID- 11430503 TI - Non-erosive reflux disease: part of the spectrum of gastro-oesophageal reflux disease, a component of functional dyspepsia, or both? AB - Over the last several decades, the incidences of gastric cancer and peptic ulcer have declined while the incidences of gastro-oesophageal reflux disease (GORD) and functional dyspepsia have reached virtually epidemic proportions. A similar trend is occurring in oesophageal cancer, with squamous cell carcinoma on the decline and adenocarcinoma on the rise, possibly due to the dramatic increase in GORD. The true clinical spectrum of these disorders, however, is only recently becoming evident: 60% of patients with GORD do not have detectable evidence of oesophagitis; they can be classified as having non-erosive or negative-endoscopy reflux disease (NERD). In this subgroup, a significant proportion will also manifest normal acid exposure on 24-h pH monitoring. Further, patients with NERD appear to be somewhat less responsive to gastric acid suppression with proton pump inhibitors. These differences, combined with the concept of the 'tender' oesophagus and the frequent presence of dyspeptic symptoms in patients with NERD, have important therapeutic implications. Therefore, considering the marked overlap in these disorders, is it realistic or clinically relevant to distinguish the entities of GORD, NERD, and functional dyspepsia? This dilemma has led to general guidelines: should heartburn predominate, treat as GORD; if dyspepsia predominates, treat as functional dyspepsia. In practical terms, each diagnosis requires consideration of the other. PMID- 11430504 TI - On-demand therapy for gastro-oesophageal reflux disease. AB - A chronic and recurrent disorder gastro-oesophageal reflux disease (GORD) affects as many as 50% of adults and is likely to require long-term therapy. Proton pump inhibitors (PPIs) are now considered first-line treatment for GORD, with daily dosing the currently accepted approach to long-term treatment. However, most patients with GORD treat themselves on an as-needed basis to relieve symptoms (primarily heartburn) associated with this disease. Limited results from clinical trials and pharmacoeconomic studies have shown that 'on-demand' therapy is both efficacious and cost-effective in patients with uncomplicated GORD. Newer PPIs, which rapidly and profoundly suppress acid secretion and provide significant symptom relief with the first dose, may be most suitable for on-demand therapy in patients with uncomplicated GORD. First-dose acid suppression and symptom relief vary among PPIs; these differences require further investigation. PMID- 11430505 TI - Gastro-oesophageal reflux disease and Helicobacter pylori or gastro-oesophageal reflux disease from Helicobacter pylori? AB - Helicobacter pylori, the most common source of chronic infection worldwide, is a principal aetiological agent of type B gastritis, gastric and duodenal ulcers, and mucosa-associated lymphoid tissue lymphomas; the presence of this pathogen is also associated with gastric carcinoma. Infection with H. pylori is prevalent in patients with gastro-oesophageal reflux disease (GORD) even though it does not appear to play an important role in GORD pathophysiology. However, epidemiological studies have shown that colonization with cagA-positive H. pylori provides significant protection against the development of GORD and its long-term complications. Whereas clinical trial results indicate that the presence of H. pylori has little influence on the effectiveness of antisecretory therapy with a proton pump inhibitor (PPI) in patients with GORD, the meta-analysis of results from long-term studies suggests that risk of GORD relapse may be reduced in the presence of H. pylori infection. Several investigators have raised concerns about increased risk for gastric neoplasia in H. pylori-positive patients treated with PPIs. However, long-term follow-up of such patients indicates neither significant risk of neoplasia nor accelerated development of gastric glandular atrophy. Thus, the excellent safety record of these compounds seems not to be compromised by the presence of H. pylori. PMID- 11430506 TI - Shortcomings of the first-generation proton pump inhibitors. AB - Proton pump inhibitors (PPIs) are widely prescribed for the treatment of gastro oesophageal reflux disease (GORD) as well as gastric and duodenal ulcers, and these agents are now considered the drugs of choice for managing such acid related disorders. Despite their well-documented efficacy and safety, first generation PPIs (omeprazole, pantoprazole and lansoprazole) have notable limitations. These drugs exhibit substantial interpatient variability in pharmacokinetics and may have significant interactions with other drugs. The time of dosing and ingestion of meals may also influence the pharmacokinetics of these agents as well as their ability to suppress gastric acid secretion. First generation PPIs also have a relatively slow onset of pharmacological action and may require several doses to achieve maximum acid suppression and symptom relief, possibly limiting their usefulness in on-demand GORD therapy. First-generation PPIs may also fail to provide 24-h suppression of gastric acid, and noctural acid breakthrough can occur even with twice-daily dosing. Both first- and second generation PPIs may be associated with adverse events consequent to gastric acid suppression, but newer PPIs have the potential to overcome some critical pharmacokinetic, pharmacodynamic, and clinical limitations of the first generation drugs. PMID- 11430507 TI - Improving on PPI-based therapy of GORD. AB - Blockade of the gastric acid pump, hydrogen-potassium adenosine triphosphatase (H+,K+-ATPase), by proton pump inhibitors (PPIs) is one of the most effective treatments for gastro-oesophageal reflux disease (GORD). In ideal terms, however, the inhibition of acid secretion should occur rapidly, on the first dose, and remain virtually complete in a dose-dependent manner. Several aspects of PPI biochemistry prevent the achievement of this ideal. PPIs target the final step of acid secretion and, due to the covalent nature of their inhibition of H+,K+ ATPase, cause suppression of acid secretion long after the drug has been eliminated. Their disadvantages stem from their mechanism of action: they require accumulation and activation in active parietal cells and have short plasma half lives, hence a relatively slow onset of action. An extension of PPI plasma half lives is an obvious goal, possibly via exploitation of probable differences in the metabolism of the two enantiomers (structural mirror images) present in current PPI formulations: e.g., clinical data on the S-enantiomer of omeprazole (esomeprazole) suggest some improvement in acid control. An alternative is to generate a pro-drug of a PPI; plasma levels of the PPI would thus depend on release of the active metabolite from the pro-drug, again extending drug half life. Another area of active investigation is the development of acid-pump antagonists to inhibit acid secretion at its final step. PMID- 11430508 TI - New-generation proton pump inhibitors: overcoming the limitations of early generation agents. AB - Proton pump inhibitors (PPIs) are now generally considered the drugs of choice for treatment of patients with serious or refractory acid-related diseases. However, despite some popular notions to the contrary, these agents are not equivalent in their pharmacokinetic, pharmacodynamic and efficacy profiles. When the two newest PPIs, rabeprazole and esomeprazole, are compared with the older drugs in this class (omeprazole, lansoprazole and pantoprazole), the newer PPIs offer several key advantages over older agents, particularly in terms of the management of gastro-oesophageal reflux disease. Rabeprazole and esomeprazole achieve more rapid and profound inhibition of acid secretion than do older agents, and they sustain this suppression to provide acid control and symptom relief over 24 h. The balanced hepatic metabolism of rabeprazole, involving both cytochrome P450 (CYP)-mediated reactions in the liver and nonenzymatic reactions, appears to confer an advantage over older PPIs in that genetic polymorphisms for CYP 2C19 do not significantly influence rabeprazole clearance and, potentially, clinical efficacy. The metabolism of esomeprazole also involves CYP 2C19, but the pharmacokinetic effects of genetic differences in the expression of this enzyme are not known. However, esomeprazole is also intended to minimize pharmacological variability. Unlike esomeprazole, the use of rabeprazole is not complicated by clinically significant drug-drug interactions of the type that have been reported for omeprazole and the wide range of medications metabolized by CYP 2C19. PMID- 11430509 TI - Reflux disease: the disorder of the third millennium. AB - Gastro-oesophageal reflux disease (GORD) affects a large group of patients and has a negative impact on quality of life. In addition, this disorder is associated with considerable long-term morbidity and mortality. In Western countries, between 10% and 30% of the population is affected by GORD. Symptoms of GORD include retrosternal burning and regurgitation, which are highly predictive of acid exposure to the oesophageal mucosa. However, erosions or ulcers in the oesophageal mucosa occur in fewer than 30% of patients with this disorder as assessed by current endoscopic techniques. Treatment of symptoms in this patient population is nonetheless of paramount importance. Current data indicate that the prevalence of GORD is increasing in the population as a whole. Although the exact reasons for this increase are unknown, changes in diet and lifestyle may play some role. Furthermore, a decreasing prevalence of Helicobacter pylori infection in the population, which is thought to exert some protection against GORD, may also be involved. Along with the increasing prevalence of GORD is an increase in GORD-related complications, including Barrett's oesophagus and oesophageal adenocarcinoma. Given the delay in onset of these GORD-associated complications, these findings are particularly alarming. As these trends continue, current techniques for the treatment of GORD will need to be refined and reevaluated in terms of quality-of-life issues, health care utilization, and cost-efficacy. Treatment of GORD in the future will require effective techniques to control symptoms, maintain remission, and prevent complications in the affected patient. PMID- 11430510 TI - Invited editorial: From 'controllable dose' to the 'next recommendations'. PMID- 11430511 TI - Invited editorial: Recent studies of the mortality and cancer morbidity experience of uranium workers and a fresh look at depleted uranium. PMID- 11430512 TI - A report on progress towards new recommendations: a communication from the International Commission on Radiological Protection. AB - Throughout the hundred-year history of the uses of ionising radiation in medicine and industry there has been advice on the need to protect people from the hazards associated with exposure. Protection standards have evolved throughout this period to reflect both the scientific understanding of the biological effects of exposure and the social and ethical standards to be applied. The Main Commission of ICRP is now considering a revised, simpler approach that is based on an individual-oriented philosophy and represents a potential shift by the Commission from the past emphasis on societal-oriented criteria. The initial proposals were promulgated through IRPA and an open literature publication was published in the Journal of Radiological Protection in June 1999. On the basis of comments received and the observations presented at the IRPA-10 Congress in May 2000, the Commission is beginning to develop the next recommendations. This article describes the issues involved in the preparation of the next recommendations and indicates the process that the Commission proposes to follow. The Commission wishes there to be an ongoing debate with an iteration of ideas over the next few years. PMID- 11430513 TI - Consultative exercise on dose assessments. AB - A summary is given of a meeting held at Sussex University, UK, in October 2000, which allowed the exchange of ideas on methods of assessment of dose to the public arising from potential authorised radioactive discharges from nuclear sites in the UK. Representatives of groups with an interest in dose assessments were invited, and hence the meeting was called the Consultative Exercise on Dose Assessments (CEDA). Although initiated and funded by the Food Standards Agency, its organisation, and the writing of the report, were overseen by an independent Chairman and Steering Group. The report contains recommendations for improvement in co-ordination between different agencies involved in assessments, on method development and on the presentation of data on assessments. These have been prepared by the Steering Group, and will be taken forward by the Food Standards Agency and other agencies in the UK. The recommendations are included in this memorandum. PMID- 11430514 TI - Risk of stillbirth in offspring of men exposed to ionising radiation. AB - Radiation genetic risk models are employed to predict the frequency of radiation related stillbirths to partners of occupationally exposed male workers, using the incidence data recently reported by Parker et al from an epidemiological study of Cumbrian births. Expanding on previously developed conservative risk estimates suggests that, of the 130 observed stillbirths to partners of male radiation workers, 0.3 cases would be attributable to paternal preconceptional irradiation, in contrast to the 17.5 (95% confidence interval: 3.1 to 31.9) cases predicted by Parker et al from their preferred dose-response model. The incompatibility of the results reported by Parker et al with those from other investigations, both epidemiological and experimental, and the inability of the study to consider a number of factors which might affect stillbirth rates, particularly those relating to the mother, make it difficult to accept that paternal irradiation received occupationally could have contributed to a detectable increase in stillbirths. PMID- 11430515 TI - A 3D Lagrangian particle model for direct plume gamma dose rate calculations. AB - A fully 3D Lagrangian particle model has been presented for calculating the direct gamma dose rates due to a radionuclide plume in the atmosphere. A continuous release of radionuclides into the atmosphere was simulated by liberating a series of puffs (each containing 100 Lagrangian particles). These puffs were released with a constant time lag between the successive puffs. The Lagrangian particle trajectories were tracked for about 25 h in a turbulent atmosphere, with a specified wind field. The atmosphere turbulent/stability characteristics like wind velocity fluctuations, eddy lifetime, etc, were obtained from the reported data in the published literature. For calculating the direct plume gamma dose rates, a point isotropic source formula has been used with appropriate attenuation and build-up factors for the air medium. Each Lagrangian particle represented a point source whose radioactive strength was calculated from the known release rate. The dose rates at ground due to the radionuclide plume were calculated by adding the contribution from each Lagrangian particle in the domain. The numerically calculated dose rates were compared with the numerical results reported in the literature. An excellent comparison was observed for downwind distances up to about 20 km. However, for distances exceeding 20 km, the numerical data were below the reported results for the Gaussian plume model. This discrepancy was due to the vertical wind shear. It is concluded that a Gaussian plume model can be used for the concentration calculations provided the lateral dispersion parameter, sigma(gamma), includes the effect of wind shear, for distances exceeding 20 km. PMID- 11430516 TI - The use of a modified technique to reduce radioactive air contamination in aerosol lung ventilation imaging. AB - The aim of this study was to reduce airborne contamination resulting from the use of aerosols in lung ventilation scintigraphy. Lung ventilation imaging is frequently performed with 99mTc-diethylenetriaminepentaacetate aerosol (DTPA), derived from a commercial nebuliser. Airborne contamination is a significant problem with this procedure; it results in exposure of staff to radiation and can reduce gamma camera performance when the ventilation is performed in the camera room. We examined the level of airborne contamination resulting from the standard technique with one of the most popular nebuliser kits and tested a modification which significantly reduced airborne contamination. Air contamination was measured while ventilating 122 patients. The modified technique reduced air contamination by a mean value of 64% (p = 0.028) compared with the standard control technique. Additionally, differences in contamination were examined when a mask or mouthpiece was used as well as differences between operators. A simplified method of monitoring air contamination is presented using a commonly available surface contamination monitor. The index so derived was proportional to air contamination (r = 0.88). The problems and regulations associated with airborne contamination are discussed. PMID- 11430517 TI - Study of radon concentrations in oil refinery premises and city dwellings. AB - Radon and its progeny concentrations were measured in several dwellings at an oil refinery premises and these concentrations were compared with those found in dwellings in Mathura and Agra cities. Radon progeny concentrations were measured using LR-115 type II nuclear track etch detectors. The radon concentrations were estimated by using a value of 0.42 for the equilibrium factor. The geometric means (GM) of radon concentrations in the refinery dwellings, Mathura city and Agra city dwellings were 97, 91 and 75 Bq m(-3) with geometric standard deviations of 1.7, 1.8 and 1.8 respectively. The average lifetime risk of lung cancer for an adjusted annual average chronic radon exposure of 69 Bq m(-3) (7.8 mWL; WL = working level) with an occupancy factor of 0.7 comes out to be 5.4 x 10(-3). PMID- 11430518 TI - A radiation incident involving detachment of a cobalt-60 industrial radiography source and its management. AB - A radiation incident took place inside an open-top radiography enclosure involving an Amertest-676 exposure device containing a cobalt-60 source of activity 2.15 TBq. The exposure device was being checked after receipt of a fresh source. The source assembly having been projected out could not be brought back into the exposure device. After thorough planning, the situation was brought under control by driving the disengaged source assembly into the exposure device. During the incident, the maximum individual whole body dose was 3.1 mSv. PMID- 11430519 TI - Healthy worker effect. PMID- 11430520 TI - Mobile phones: developments in the UK. PMID- 11430521 TI - Draft Euratom Directive on the Control of High Activity Sealed Radioactive Sources (HAAS). PMID- 11430522 TI - IAEA Meeting: International Conference of National Regulatory Authorities with Competence in the Saftey of Radiation Sources and the Security of Radioactive Materials. PMID- 11430523 TI - In memoriam to Dov Front. PMID- 11430524 TI - Remembering Dov Front. PMID- 11430525 TI - The value of Ga-67 scintigraphy and F-18 fluorodeoxyglucose positron emission tomography in staging and monitoring the response of lymphoma to treatment. AB - Gallium-67 scintigraphy (GS) has the ability to provide important diagnostic and prognostic information for the evaluation of patients with lymphoma. GS is superior to morphologic imaging techniques because of its affinity to viable lymphoma cells. The value of GS lies not in the initial diagnosis but primarily in assessing the results of treatment and in the follow-up of patients with lymphoma. Nevertheless, GS has not gained the expected wide acceptance, possibly because of the meticulous technique required and the expertise needed for optimal interpretation. The introduction of positron emission tomography (PET) with F-18 fluorodeoxyglucose (FDG) as a tumor-seeking agent, which provides images of superior quality, may have an impact on the current role of GS in the management of patients with lymphoma. FDG-PET seems to share with GS the advantages of a tumor viability agent. It appears to be more sensitive for detecting nodal and extranodal sites of disease than GS and may have predictive value during and after therapy for lymphoma. These potential clinical and economic advantages of FDG-PET need to be confirmed in systematic, large-scale prospective studies. PMID- 11430526 TI - The fusion of anatomic and physiologic imaging in the management of patients with cancer. AB - Imaging is of major clinical importance in the noninvasive evaluation and management of patients with cancer. Computed tomography (CT) and other anatomic imaging modalities, such as magnetic resonance imaging (MRI) or ultrasound, have a high diagnostic ability by visualizing lesion morphology and by providing the exact localization of malignant sites. Nuclear medicine provides information on the function and metabolism of cancer. Over the last decade, there have been numerous attempts to combine data obtained from different imaging techniques. Fused images of nuclear medicine and CT (or to a lesser extent, MRI) overcome the inherent limitations of both modalities. Valuable physiologic information benefits from a precise topographic localization. Coregistered data have been shown to be useful in the evaluation of patients with cancer at diagnosis and staging, in monitoring the response to treatment, and during follow up, for early detection of recurrence. Time-consuming and difficult realignment and computation for fusion of independent studies have, until now, limited the use of registration techniques to pilot studies performed in a small number of patients. The development of the new technology of single photon emission computed tomography/CT and positron emission tomography/CT that allows for combined functional and anatomic data acquisition has the potential to make fusion an everyday clinical tool. PMID- 11430527 TI - The role of nuclear medicine in monitoring treatment in skeletal malignancy. AB - The nuclear medicine bone scan has historically been one of the most common investigations to stage and monitor skeletal malignancy. Current guidelines for using radiographs to assess the response of skeletal metastases to systemic therapy are limited in their ability to give a timely result. Despite some minor limitations caused by the flare phenomenon, skeletal scintigraphy remains widely used for this purpose, both clinically and in trials of new cancer treatments. Nuclear medicine has also played an important role in the posttherapy evaluation of primary bone tumors, both with bone agents and nonspecific tumor agents, such as 201Tl. In the future, it is possible that positron emission tomography radiopharmaceuticals such as 18F-fluorodeoxyglucose may prove to be superior in predicting and measuring treatment response in primary and metastatic bone and bone marrow disease, but further work is required in this area. PMID- 11430528 TI - Monitoring treatment for osteoporosis by using bone densitometry. AB - In recent years, the range of treatments available for patients with osteoporosis has greatly increased. A decade ago, the only proven therapy was estrogen, but today the choices include bisphosphonates, selective estrogen-receptor modulators, calcitonin, and parathyroid hormone. Clinical trials involving bone mineral density (BMD) scans of the spine and femur have had an important role in the evaluation of these new therapies. In particular, the efficacy of treatments has now been verified in large studies powered to show reductions in fracture risk. In contrast with the use of BMD scans in research studies, their value for monitoring response to treatment in individual patients is less certain. In many cases, the increases in BMD are too small to reliably distinguish between true changes and measurement error. However, experience with estrogen and bisphosphonates suggests that these treatments have a beneficial effect on bone in most patients. Follow-up scans for patients taking these agents are therefore of limited value, apart from offering reassurance to the patient and the doctor. However, when new therapies are first introduced, follow-up scans have a useful role in building up the physician's experience and confidence. PMID- 11430529 TI - Cardiac nuclear medicine in monitoring patients with coronary heart disease. AB - Patient monitoring is a major indication for cardiac nuclear medicine procedures. Stress myocardial perfusion scintigraphy was initially used for diagnosis, but monitoring patients with coronary artery disease has become more common. Stress myocardial perfusion scintigraphy has been shown to provide a considerable amount of incremental prognostic information, which may be useful in selecting patients for therapy. In patients being considered for revascularization, fluorodeoxyglucose can be used to identify regions of dysfunctional but viable myocardium, even within regions that show fixed defects on stress perfusion imaging. It can be used to select a group of patients who will improve function with revascularization and who may have an improved outcome. Thus, cardiac nuclear medicine plays a pivotal role in monitoring patients with coronary artery disease. PMID- 11430530 TI - Nuclear endocrinology as a monitoring tool. AB - Malignant endocrine disorders have been an enigma over the last few decades, from genetic, clinical, and imaging perspectives. The detection of the primary tumor and the identification of recurrent disease have been essentially based on various anatomic techniques, with localization procedures extensively developed for staging, follow-up, radio-guided surgery, and therapy. Frequently, the lesions are too small to cause anatomic alterations, or they are obscured by the changes in anatomic planes that occur after initial surgery. Small lesions, however, are the ones that can potentially be cured. Thus, every attempt should be made to localize these sites before further growth and dissemination occur beyond the scope of cure. Since the advent of iodine-131 for staging and follow up of patients with differentiated thyroid carcinoma, the search has led to the use of radioiodinated metaiodobenzylguanidine (MIBG) for recurrent pheochromocytoma and neuroblastoma, to the development of antibodies to carcinoembryonic antigen for the staging and treatment of medullary thyroid carcinoma, and to the characterization of peptide receptors on neuroendocrine tumors. Additionally, there has been a breakthrough with the use of positron emitters in nuclear oncology, including F-18-fluorodeoxyglucose, for I-131 negative metastases of differentiated thyroid carcinoma, recurrent medullary thyroid carcinoma, malignant pheochromocytoma, and adrenocortical carcinoma. Undoubtedly, optimal care of the patient requires both the expertise of the treating endocrinologist and the use of various imaging techniques in the diagnosis, staging, and follow-up of these diseases. PMID- 11430531 TI - Cold vertebrae on Ga-67 scintigraphy. PMID- 11430532 TI - Diffuse liver and spleen accumulation on Tc-99m methylene diphosphonate bone scan. PMID- 11430533 TI - Noninvasive ventilation routine therapy for community-acquired pneumonia? Not so fast! PMID- 11430534 TI - Antagonization of tumor necrosis factor in snake bite. A new approach for an old threat. PMID- 11430535 TI - Pathogenesis and treatment of renal dysfunction in rhabdomyolysis. AB - Rhabdomyolysis is a major cause of acute renal failure, and recent experimental data have provided a better understanding of the pathophysiology of the renal dysfunction. Renal failure is due to renal vasoconstriction, tubular damage caused by oxidant injury, and possibly tubular obstruction. Recent studies have provided greater insight into the rationale behind current therapy and potential treatment strategies. This review thus aims to summarise current understanding of the causes, pathogenesis and treatment of renal failure caused by rhabdomyolysis. PMID- 11430536 TI - Non-invasive pressure support ventilation in severe community-acquired pneumonia. AB - OBJECTIVE: To explore three aspects of non-invasive pressure support ventilation (NIPSV) applied by face mask to patients with acute respiratory failure (ARF) due to severe community-acquired pneumonia (CAP): (1) the initial acute effects on respiratory rate, gas exchange and hemodynamics, (2) the clinical course and outcome during ICU and hospital stay, (3) the nursing workload as measured by the daily PRN 87 (Project Research in Nursing) score. SETTING: Medical ICU, University Hospital. DESIGN: Prospective, observational study. PATIENTS: Patients without any prior history of chronic lung disease, consecutively admitted to the ICU to receive NIPSV for ARF due to severe CAP. MEASUREMENTS AND RESULTS: (means +/- SD): Twenty-four patients aged 49+/-17 years, admission APACHE II 13+/-5, were included. Admission PaO2/FIO2, alveolar-arterial oxygen difference (DA-aO2) and PaCO2 were 104+/-48, 447+/-120 and 40+/-10 mmHg, respectively. All patients were normotensive. During the initial NIPSV trial respiratory rate decreased from 34+/-8 to 28+/-10 breaths/min (p < 0.001) and arterial oxygenation improved (PaO2/FIO2 104+/-48 vs 153+/-49, DA-aO2 447+/-120 vs 370+/-180 mmHg, p < 0.001) while PaCO2 remained unchanged. There were no hemodynamic effects. Subsequently, a total of 133 NIPSV trials were performed (median duration 55 min, range 30-540 min) over 1-7 days. No complication occurred during NIPSV. Sixteen patients were intubated (66%) 1.3+/-1 days after inclusion. Upon inclusion, the patients who were subsequently intubated were older (55+/-15 vs 37+/-12 years) and more severely hypoxemic (63+/-11 vs 80+/-15 mmHg, p < 0.05) than those not requiring intubation. Eight patients died (33 %), all in the intubated group. Median lengths of stay in the ICU and hospital were longer in intubated patients (ICU 16 days, range 3-64 vs 6 days, range 3-7, p < 0.05; hospital 23 days, range 9-77 vs 9.5 days, range 4-42, p < 0.05). Mean daily total PRN points were stable throughout the NIPSV period and were not different between the groups. Only 14% of PRN points resulted from respiratory therapy interventions. PRN score was higher during the first 24 h following intubation than during the first 24 h of NIPSV (278+/-55 vs 228+/-24 points, p < 0.05). CONCLUSION: Despite initial improvement in arterial oxygenation with NIPSV in patients with ARF due to severe CAP, the intubation rate is high. However, the more favorable outcome and shorter ICU and hospital stays when intubation is avoided, as well as the short delay required to assess the success or failure of NIPSV warrants a trial of NIPSV in this setting. The nursing workload remains stable during NIPSV and does not result predominantly from respiratory therapy interventions. PMID- 11430537 TI - Neostigmine resolves critical illness-related colonic ileus in intensive care patients with multiple organ failure--a prospective, double-blind, placebo controlled trial. AB - OBJECTIVE: Critical illness-related colonic ileus (CIRCI) is characterized by the non-passage of stools in critically ill patients as a result of the absence of prokinetic movements of the colon, while the upper gastrointestinal tract functions properly and mechanical ileus is absent: We investigated whether neostigmine resulted in defecation in patients with CIRCI. DESIGN: Double blinded, placebo-controlled prospective study. SETTING: Eighteen-bed intensive care unit. PATIENTS: Thirty ventilated patients with multiple organ failure with CIRCI for > 3 days. INTERVENTION: Continuous intravenous administration of neostigmine 0.4-0.8 mg/h over 24 h, or placebo. MEASUREMENTS AND RESULTS: Time to first defecation and adverse reactions were recorded. Thirty patients were randomized, 24 could be evaluated. The mean prestudy time was 5 days, mean APACHE II score on admission was 23.2, and mean MOF score on the day of the study was 6.4. Of the 13 patients receiving neostigmine, 11 passed stools, whereas none of the placebo-treated patients passed stools (P < 0.001). After 24 h, the non responders received in a cross-over fashion neostigmine or placebo respectively. Eight out of the 11 neostigmine patients now passed stools (mean 11.4 h), and none of the placebo patients. Overall, in none of the patients did passage of stools occur during placebo infusion, whereas 19 of the 24 neostigmine-treated patients had defecation (79%). No acute serious adverse effects occurred, but three patients had ischemic colonic complications 7-10 days after treatment. CONCLUSION: Continuous infusion of 0.4-0.8 mg/h of neostigmine promotes defecation in ICU patients with a colonic ileus without important adverse reactions. PMID- 11430538 TI - The value of the chloride: sodium ratio in differentiating the aetiology of metabolic acidosis. AB - OBJECTIVE: Stewart's physicochemical approach to acid-base balance defines the aetiology of a metabolic acidosis by quantifying anions of tissue acids (TA), which consist of unmeasured anions (UMA) and/or lactate. We hypothesised that an increase in TA during metabolic acidosis would lead to a compensatory fall in the plasma chloride (Cl) relative to sodium (Cl:Na ratio) in order to preserve electro-neutrality. Thus, the Cl:Na ratio could be used as a simple alternative to the anion gap in identifying raised TA. PATIENTS: Two hundred and eighty two consecutive patients who were admitted to our Paediatric Intensive Care were enrolled in the study. INTERVENTIONS: We obtained 540 samples (admission n = 282, 24 h n = 258) for analysis of blood chemistry, lactate and quantification of TA and UMA. Samples were subgrouped into those with metabolic acidosis (standard bicarbonate < 22 mmol/l) either with or without increased UMA (> 3 mEq/l). MEASUREMENTS AND RESULTS: Metabolic acidosis occurred in 46% of samples, of which 52.3% (120/230) had increased UMA. The dominant component of TA was UMA rather than lactate, and these two components did not always rise in tandem. Our hypothesis of relative hypochloraemia was supported by a lower Cl:Na ratio (P < 0.0001) but not a lower absolute Cl (P = 0.5) in the acidotic subgroup with raised UMA, and by the inverse relationship between TA and the Cl:Na ratio. (coefficient of determination (r2) = 0.37, P < 0.0001). The best discriminator for the presence of raised TA was the albumin-corrected anion gap (AGcorr), however, this could not track changes in TA with clinical accuracy. The Cl:Na ratio discriminated reasonably well, a ratio of < 0.75 identified TA (positive predictive value (PPV) 88%) with a likelihood ratio (LR) similar to the AG (7.8 vs7.4). Conversely, a high ratio (> 0.79) excluded TA (PPV 81%, LR 4.5). Base deficit (BD) and lactate performed poorly. CONCLUSION: In metabolic acidosis due to TA, plasma Cl concentration decreases relative to sodium. The Cl:Na ratio is a simple alternative to the AG for detecting TA in this setting. PMID- 11430539 TI - Growth hormone does not affect albumin synthesis in the critically ill. AB - OBJECTIVE: To study the effect of growth hormone (GH) on albumin synthesis in critically ill patients. DESIGN: Prospective randomized controlled study. SETTING: Two intensive care units, university hospital and county hospital, respectively. PATIENTS: Twenty-two critically ill patients in the intensive care unit. INTERVENTIONS: Albumin synthesis was measured twice in each patient, with a 5-day interval. The patients in the control group (n = 11) received standard intensive care unit (ICU) treatment between measurements, whereas those in the GH group (n = 11) also received 0.3 U/kg daily of human recombinant GH. MEASUREMENTS AND RESULTS: Albumin synthesis was measured by labeling with L [2H5]phenylalanine. In the control group, the fractional synthesis rate (FSR) of albumin was 16.3+/-4.1%/day (mean and SD) in the first measurement and 15.7+/ 4.2%/day 5 days later (NS), whereas in the GH group the corresponding values were 17.0+/-4.7%/day and 16.7+/-5.5%/day (NS). The calculated absolute synthesis rates of albumin, based on FSR and intravascular albumin mass, also showed no effect of GH. CONCLUSION: Albumin synthesis rates were consistently higher in the two groups of critically ill patients than previously reported values in healthy subjects. However, GH treatment for 5 days neither stimulated nor inhibited albumin synthesis rates in these critically ill patients. PMID- 11430540 TI - Albumin supplementation during the first week after a burn does not mobilise tissue oedema in humans. AB - OBJECTIVE: To measure water balance and changes in distribution, and the effect of giving supplementary albumin, early after a burn injury. DESIGN: Consecutive patients (matched groups) and healthy controls. SETTING: National burn unit in a Swedish university hospital. PATIENTS AND SUBJECTS: Eighteen patients with 18% 90% total burned surface area and 16 healthy male control subjects. INTERVENTIONS: The patients were given an intravenous infusion of ethanol over 1 h, 0.35-0.60 g/kg body weight, and a bolus of 3.3 to 6.5 g of iohexol. The control subjects were given the same amounts of either ethanol or iohexol. Patients were subdivided into two groups according to whether or not they received supplementary albumin starting 12 h post-burn. MEASUREMENTS AND RESULTS: Blood samples were drawn at 20-30 min intervals over 4 h after the start of the infusion. Serum ethanol was measured by headspace gas chromatography, and iohexol with high-pressure liquid chromatography (HPLC). Distribution volume was calculated from the concentration-time profiles. Total body water (TBW) was measured by the ethanol tracer and bioelectric impedance (BIA) techniques, and estimated extracellular water (ECWest) by iohexol tracer. They were all significantly increased after a burn. Excess water was accumulated mainly in the extracellular compartment. It declined towards normal values (those of volunteers) at the end of the week. Albumin supplementation did not influence the amount or distribution of the excess fluid. CONCLUSION: Body water increases after a burn. Excess water is mainly deposited in the extracellular space. Tissue oedema fluid is not mobilised by albumin supplementation. PMID- 11430541 TI - Validating the Sedation-Agitation Scale with the Bispectral Index and Visual Analog Scale in adult ICU patients after cardiac surgery. AB - OBJECTIVE: To validate the Sedation-Agitation Scale (SAS) with the Visual Analog Scale (VAS) and Bispectral Index (BIS) in adult ICU patients after cardiac surgery. DESIGN: Prospective study comparing blinded evaluations of the SAS, VAS and BIS. SETTING: Forty-two-bed multidisciplinary ICU. PATIENTS AND PARTICIPANTS: Convenience sample of 39 adults after cardiac surgery. MEASUREMENTS AND RESULTS: Bispectral Index 3.2 was continuously recorded using the Aspect A-1000 and evaluators were blinded to this value. The bedside nurse and a trained researcher independently rated wakefulness using a 100 mm VAS upon patient arrival on the ICU, at first awakening, when ventilator weaning was started and after extubation; the researcher also evaluated patients using SAS. Upon arrival on the ICU, the median SAS score was 2 (interquartile range = 1-3), the mean VAS was 26+/-30 and the mean BIS was 70+/-16. Twenty-four patients underwent a trial of weaning from mechanical ventilation with a SAS of 4 (IQR = 4), VAS of 86+/-12 and BIS of 87+/-10. SAS correlated well with VAS performed by one researcher (r = 0.91, p < 0.001) or by 19 different bedside nurses (r = 0.43, p < 0.001) and with BIS 3.2 (r = 0.60, p < 0.001). The correlation between SAS and BIS was reduced in patients with above average electromyogram (EMG) power. As a measure of construct validity, significant differences were noted for the BIS, SAS, VAS and EMG between ICU arrival and extubation (all p < 0.001). CONCLUSIONS: Sedation Agitation Scale and BIS are valid measures of wakefulness after cardiac surgery, but EMG interference may affect the accuracy of BIS for a small percentage of patients not receiving neuromuscular blockade. PMID- 11430542 TI - Intensive Care Delirium Screening Checklist: evaluation of a new screening tool. AB - OBJECTIVE: Delirium in the intensive care unit is poorly defined. Clinical evaluation is difficult in the setting of unstable, often intubated patients. A screening tool may improve the detection of delirium. METHOD: We created a screening checklist of eight items based on DSM criteria and features of delirium: altered level of consciousness, inattention, disorientation, hallucination or delusion, psychomotor agitation or retardation, inappropriate mood or speech, sleep/wake cycle disturbance, and symptom fluctuation. During 3 months, all patients admitted to a busy medical/surgical intensive care unit were evaluated, and the scale score was compared to a psychiatric evaluation. RESULTS: In 93 patients studied, 15 developed delirium. Fourteen (93%) of them had a score of 4 points or more. This score was also present in 15 (19%) of patients without delirium, 14 of whom had a known psychiatric illness, dementia, a structural neurological abnormality or encephalopathy. A ROC analysis was used to determine the sensitivity and specificity of the screening tool. The area under the ROC curve is 0.9017. Predicted sensitivity is 99% and specificity is 64%. CONCLUSION: This study suggests that the Intensive Care Delirium Screening Checklist can easily be applied by a clinician or a nurse in a busy critical care setting to screen all patients even when communication is compromised. The tool can be utilized quickly and helps to identify delirious patients. Earlier diagnosis may lead to earlier intervention and better patient care. PMID- 11430543 TI - The impact of organisational change on outcome in an intensive care unit in the United Kingdom. AB - OBJECTIVES: To study the change in outcome for patients admitted to an intensive care unit following the establishment of a team of resident medical staff and a change from an "open" to a "closed" organisational format. DESIGN: Database review of prospectively collected data. SETTING: Intensive care unit of a postgraduate teaching hospital. SUBJECTS: 1134 admissions to the intensive care unit over a 3-year period, of whom 476 (42%) followed elective surgery. MAIN OUTCOME MEASURE: Hospital mortality corrected for illness severity by using the APACHE II scoring system. RESULTS: Crude hospital mortality fell from 28% before the changes to 20% afterwards (P = 0.01). With correction for case-mix factors, the probability of death after the changes was reduced by almost half (OR 0.51; CI 0.32, 0.82, P = 0.005). CONCLUSION: A "closed" format of organisation of the delivery of care may result in improved outcomes for patients admitted to intensive care units. PMID- 11430544 TI - Statistical models to predict the need for postoperative intensive care and hospitalization in pediatric surgical patients. AB - OBJECTIVE: To develop statistical models for predicting postoperative hospital and ICU stay in pediatric surgical patients based on preoperative clinical characteristics and operative factors related to the degree of surgical stress. We hypothesized that preoperative and operative factors will predict the need for ICU admission and may be used to forecast the length of ICU stay or postoperative hospital stay. DESIGN: Prospective data collection from 1,763 patients. SETTING: Tertiary care children's hospital. PATIENTS AND PARTICIPANTS: All pediatric surgical patients, including those undergoing day surgery. Patients undergoing dental or ophthalmologic surgical procedures were excluded. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: A logistic regression model predicting ICU admission was developed from all patients. Poissonregression models were developed from 1,161 randomly selected patients and validated from the remaining 602 patients. The logistic regression model for ICU admission was highlypredictive (area under the receiver operating characteristics (ROC) curve = 0.981). In the data set used for development of Poisson regression models, significant correlations occurred between the observed and predicted ICU stay (Pearson r = 0.468, p < 0.0001, n = 131) and between the observed and predicted hospital stay for patients undergoing general (r = 0.695, p < 0.0001), orthopedic (r = 0.717, p < 0.0001), cardiothoracic (r = 0.746, p < 0.0001), urologic (r = 0.458, p < 0.0001), otorhinolaryngologic (r = 0.962, p < 0.0001), neurosurgical (r = 0.7084, p < 0.0001) and plastic surgical (r = 0.854, p < 0.0001) procedures. In the validation data set, correlations between predicted and observed hospital stay were significant for general (p < 0.0001), orthopedic (p < 0.0001), cardiothoracic (p = 0.0321) and urologic surgery (p = 0.0383). The Poisson models for length of ICU stay, otorhinolaryngology, neurosurgery or plastic surgery could not be validated because of small numbers of patients. CONCLUSIONS: Preoperative and operative factors may be used to develop statistical models predicting the need for ICU admission in pediatric surgical patients, and hospital stay following general surgical, orthopedic, cardiothoracic and urologic procedures. These statistical models need to be refined and validatedfurther, perhaps using data collection from multiple institutions. PMID- 11430545 TI - Antagonization of TNF attenuates systemic hemodynamic manifestations of envenomation in a rat model of Vipera aspis snakebite. AB - OBJECTIVES: Tumor necrosis factor (TNF) has been reported as a mediator of local tissue injury following snake envenomation in an intact rat model. We investigated whether systemic release of TNF occurs following Vipera aspis envenomation. We further analyzed the possible connection between envenomation related hemodynamic depression and TNF antagonization (TNF antibodies or soluble TNF receptor). DESIGN: A prospective, randomized, controlled experimental study using a rat model for snake envenomation. SETTINGS: A medical university hospital research laboratory. INTERVENTION: Eighty rats (300-400 g) were divided into four groups (n = 20): control and three experimental groups. Intramuscular injection of V. asis 500 microg/kg was administered to the three experimental groups: venom only (group 1), venom and 40 microg anti-TNF antibodies (group 2), venom and 250 microg soluble TNF receptor (p55-R; group 3). Hemodynamic parameters were monitored up to 4 h following venom injection. MEASUREMENTS AND RESULTS: A significant hemodynamic deterioration (reduction in heart rate and blood pressure) occurred 30 min following venom injection in group 1 compared to groups 2 and 3, where hemodynamic parameters remained stable throughout the 4 h observation period. Serum levels of TNF were detected 15 min after venom injection and peaked after 2 h at 485+/-12 pg/ml. CONCLUSIONS: The hemodynamic consequences of intramuscular injection of V. aspis venom can be blunted in a rat by systemic antagonization of TNF activity prior to venom injection. The poisonous hemodynamic effects of the V. aspis venom might be caused by systemic release of TNF. PMID- 11430546 TI - Effects of inhalation of perfluorocarbon aerosol on oxygenation and pulmonary function compared to PGI2 inhalation in a sheep model of oleic acid-induced lung injury. AB - OBJECTIVE: To evaluate the effects of PFC aerosol compared to PGI2 aerosol and NaCl aerosol on gas exchange and lung mechanics in oleic acid-induced acute lung injury. DESIGN: A prospective, controlled, randomised, in vivo animal laboratory study. SETTING: Research laboratory at an university hospital. SUBJECTS: Twenty one (n = 21) adult sheep of either gender weighing 26.8+/-6.4 kg. INTERVENTIONS: The animals were randomised to three groups: PFC aerosol (perfluorooctane), PFC group; prostacyclin aerosol (Flolan), PGI2 group; and NaCl aerosol (0.9% sodium chloride solution), control group. After induction of anaesthesia and placement of vascular catheters, lung injury was induced with 0.12 ml x kg(-1) oleic acid. Aerosols were continuously administered for 2 h using a jet nebuliser. Gas exchange, pulmonary mechanic, and haemodynamic parameters were obtained at regular intervals. MEASUREMENTS AND MAIN RESULTS: PFC aerosol increased oxygenation (PaO2) 15 min after the initiation of treatment up to 120 min (P < 0.05). Transpulmonary shunt improved in the PFC group (P < 0.05) while it did not change in the two other groups. PFC aerosol reduced maximum airway pressure (Pmax) (median) significantly from (median) 38 mbar to 32 mbar (P < 0.05). Static compliance improved significantly in the PFC group (P < 0.05). CONCLUSION: The inhalation of a PFC aerosol led to a significant improvement in pulmonary mechanics and gas exchange, which was not observed in the other two groups. These data suggest that a small dose of perfluorocarbon will have beneficial effects on gas exchange and respiratory mechanics. Therefore, the non-invasive aerosol application technique seems to be a reasonable alternative to administer perfluorocarbons in severe lung injury. PMID- 11430547 TI - Volume dependence of respiratory system resistance during artificial ventilation in rabbits. AB - The volume dependence of respiratory resistance (Rrs), usually observed during normal breathing, is expected to be accentuated during expiratory flow limitation (EFL). In order to quantify this dependence we studied the pressure, flow, and volume data obtained from eight New Zealand rabbits, artificially ventilated at different levels of applied expiratory pressure (0-10 hPa), before and during histamine i. v. infusion. EFL was provoked by lowering the expiratory pressure and was detected by the application of an additional negative expiratory pressure and by forced oscillations. The analysis of respiratory system mechanics was performed by multiple regression, using the classical linear first-order model and also a nonlinear model, accounting for volume dependence of Rrs. Both models satisfactorily fitted the data in the absence of EFL. The nonlinear model proved to be more appropriate in the presence of EFL. The coefficient expressing the volume dependence of Rrs (Rvd) was significantly more negative during EFL. Rvd values were highly correlated with the fraction of the tidal volume left to be expired at the onset of EFL. A threshold Rvd value of -1,000 (hPa x s x l(-2)) detected EFL with high sensitivity and specificity. We conclude that a strongly negative volume dependence of Rrs is a reliable and noninvasive index of EFL during artificial ventilation. PMID- 11430548 TI - Hypoxaemic reperfusion ameliorates the histopathological changes in the pig brain after a severe global cerebral ischaemic insult. AB - BACKGROUND AND PURPOSE: We have recently shown that hypoxaemic reperfusion, after an ischaemic brain insult, improves neurological outcome and decreases lipid peroxidation. In the present study, we investigated the effect of hypoxaemic reperfusion on brain histopathological changes. METHODS: Sixteen pigs subjected to a 10-min global cerebral ischaemia were either hypoxaemically (PaO2 = 35 mmHg, hypoxaemic reperfusion (HR) group, n = 8) or hyperoxaemically (PaO2 > 300 mmHg, control (C) group, n = 8) reperfused. The brains were removed 24 h after reperfusion and six neuropathological abnormalities were evaluated blindly and scored semi-quantitatively (0: normal to 3: severe injury) on eight representative regions of the brain. The overall cumulative score of the abnormalities and their regional prevalence, as well as the neurological outcome, were compared between the two groups. RESULTS: The neuronal degeneration, assessed in terms of cumulative score (P = 0.002) and regional prevalence (P = 0.025 to P = 0.041), was lower in the HR group than in the C group. Spongy degeneration attained statistically significant difference only in cerebellum (P = 0.002) and inflammation only in hippocampus (P = 0.046) but the difference in the cumulative score of these abnormalities was not statistically significant. The difference of the three neurological assessments over time was statistically significant between the two groups, i.e. after resuscitation (P = 0.001), at 8 h (P = 0.006) and at 24 h (P = 0.001) after reperfusion. CONCLUSIONS: Hypoxaemic reperfusion during resuscitation from a severe global ischaemic cerebral insult is associated with statistically significantly fewer histopathological changes of the brain than in controls. This is associated with a superior neurological outcome. PMID- 11430549 TI - Effects of tidal volume reduction in acute respiratory distress syndrome on gastric mucosal perfusion. AB - OBJECTIVE: This study was conducted with the aim of testing the effects of a reduction in tidal volume (V(T)) on gastric mucosal perfusion using laser-Doppler flowmetry in patients with acute respiratory distress syndrome (ARDS). DESIGN: It was designed as a prospective study. PATIENTS: Seventeen patients with ARDS were enrolled in the study. All patients were mechanically ventilated in volume controlled mode. Before the start of the protocol, V(T) was set at 9 ml/kg body weight. INTERVENTION: V(T) was reduced to 6 ml/kg body weight. MEASUREMENTS AND RESULTS: Measurements of systemic hemodynamic parameters and gastric mucosal blood flow (GMBF) were obtained before and after reduction of V(T). Cardiac index, heaart rate and pulmonary arterial pressure increased significantly after V(T) reduction. The increase in cardiac output was observed in all patients. However, despite a mean 25% increase in cardiac output after V(T) reduction, no significant increase in mean GMBF was observed, and individual GMBF responses were heterogeneous. CONCLUSION: V(T) reduction in patients with ARDS, despite resulting in an increase in cardiac output, did not change gastric mucosal perfusion. The heterogeneity in the individual response of GMBF to V(T) reduction could be due to opposite direct (i.e., local vasodilatory effect) and indirect (i.e., global sympathetic stimulation) effects of hypercapnia on gut vessels. PMID- 11430550 TI - Value of Candida antigen and antibody assays for the diagnosis of invasive candidosis in surgical intensive care patients. AB - During a 3-year period, a clinical diagnosis of invasive candidosis was made in 8 out of 2054 consecutive surgical intensive care unit (ICU) patients. These patients were retrospectively matched with 16 control patients who underwent similar surgical procedures and had a similar clinical course except for negative Candida cultures. In all patients, Candida antigen (Ramco CandTec serum antigen test) and antibody serology (Candida HA test) were determined at least once a week during their stay. The antigen test was positive in 1/8 patients and 4/16 controls and thus did not differentiate patients with candidosis from non infected controls. The HA antibody titer results fulfilled the manufacturer's criteria for positivity in 7/8 patients with candidosis and 2/16 control patients. Thus, the Candida HA antibody test, but not the Ramco antigen test, can be recommended to confirm a clinical diagnosis of invasive candidosis. PMID- 11430551 TI - Tonicity balance, and not electrolyte-free water calculations, more accurately guides therapy for acute changes in natremia. AB - The usual way to decide why hyponatremia or hypernatremia has developed and to plan goals for its therapy is to analyze events in electrolyte-free water (EFW) terms. We shall demonstrate that an EFW balance does not supply this information. Rather, one must calculate mass balances for water and sodium plus potassium separately (a tonicity balance) to understand the basis for the change in natremia and the proper goals for its therapy. These points are illustrated with a clinical example. PMID- 11430552 TI - Determination of plasma volume by indocyanine green--validation of the method and use in patients after cardiopulmonary bypass. AB - OBJECTIVE: Validation of plasma volume (PV) determination by indocyanine green (ICG) in comparison to the gold-standard method with radioiodinated albumin, and investigation of the effect of commonly used plasma expanders (albumin, hydroxyethyl starch, and polygelatine) on PV in the early postoperative phase in patients undergoing cardiac surgery. DESIGN: Prospective clinical study. SETTING: Department of medicine and intensive care unit at a university hospital. PATIENTS AND PARTICIPANTS: Ten healthy volunteers and 21 patients after elective open heart surgery. MEASUREMENTS AND RESULTS: PV of subjects was measured by i.v. injecting 5 microCi [125I]albumin (I-ALB). One hour later, PV was determined by a peripheral i. v. injection of 0.25 mg/kg body weight ICG (ICG1). In five subjects PV was measured repeatedly by ICG (ICG2) 1 h after ICG1. Mean PV of I-ALB and ICG1 or ICG2 showed consistent results. Further, we investigated central vs peripheral intravenous injection of ICG in six patients after open-heart surgery compared to [125I]albumin. There was no difference between mean PV measured by [125I]albumin and peripheral ICG (P = 0.40). PV determined by central injection of ICG was significantly higher than by the other methods. In 15 patients PV was determined by [125I]albumin. Thereafter, patients were randomly divided into three groups. Group ALB was infused with 1.75 ml/kg body weight human albumin 20%, group HAES with 5.25 ml/kg body weight hydroxyethyl starch 6%, and group HAEM with 7.0 ml/kg body weight polygelatine 3.5%. PV was measured 1 h and 4 h after infusion by ICG. There were no significant changes in PV between the groups. CONCLUSIONS: PV determination by peripheral i. v. injection of ICG produced reliable and consistent results when a reactive hyperaemia was produced by a tourniquet prior to injection. Therefore, central venous injection of ICG may not be prerequisite for precise measurements of PV. The expected acute increase in PV after infusion of commonly used plasma expanders after cardiac surgery was not found. PMID- 11430553 TI - Acute amitraz intoxication in human. AB - OBJECTIVE: To review the clinical profiles and therapies instituted for patients with acute amitraz intoxication. DESIGN: Retrospective study. SETTING: Multidisciplinary intensive care unit (ICU) of a university hospital. PATIENTS: Ten patients admitted to the ICU of Ege University Hospital. RESULTS: Ten patients between 4 and 34 years of age were evaluated. The intoxication was the result of a suicide attempt in five cases and accidental in the rest. Altered consciousness, nausea, vomiting and dizziness were the common initial symptoms. The major clinical findings in the ICU were somnolence, coma, miosis, mydriasis, bradycardia, respiratory failure requiring mechanical ventilation, and hypo- and hyperthermia. Blood glucose level was increased significantly in three cases and urinary output was increased in two. The length of stay was between 2 and 5 days. CONCLUSION: The signs and symptoms of acute amitraz intoxication appear rapidly and become established within hours. The initially severe clinical situation improves fast. Cases of this rare mode of intoxication should be carefully followed and treated in the ICU. PMID- 11430554 TI - Extracorporeal circulation for acute respiratory failure. PMID- 11430555 TI - A simple intervention to improve satisfaction in patients and relatives. PMID- 11430556 TI - Hypoxemic acute respiratory failure of severe tetanus: is it of cardiac or pulmonary origin? PMID- 11430557 TI - Gastric rupture after nasopharyngeal oxygen administration. PMID- 11430558 TI - Impending complete rupture of proximal aortic dissection manifested by ST-segment elevation. PMID- 11430559 TI - Failure of intrapericardial streptokinase in purulent pericarditis. PMID- 11430560 TI - Simultaneous side effects of both clozapine and valproate. PMID- 11430561 TI - Comatose state due to severe hyponatremia in a patient with the syndrome of inappropriate antidiuretic hormone secretion. PMID- 11430562 TI - Brain death: clear definitions and terminology. PMID- 11430563 TI - Hemophagocytic syndrome in the critically ill. PMID- 11430564 TI - Salmonella infection with multi-organ failure precipitated by trauma. PMID- 11430565 TI - The diurnal rhythm of adrenocorticotropic hormone secretion in the assessment of the adequacy of replacement therapy in primary chronic adrenal failure. AB - Utilization of the daily rhythm of ACTH secretion was assessed as a measure of the adequacy of replacement therapy in primary chronic adrenal failure (PCAF: Addison's disease) with glucocorticoids (GC) by assaying plasma ACTH by radioimmunoassay every 4 h for one day in three groups of patients. Patients of group 1 had PCAF (n = 14) and received replacement therapy consisting of prednisolone (5 mg at 09:00 and 2.5 mg at 14:00) and group 2 patients received dexamethasone (0.5 mg at 23:00) in combination with prednisolone (2.5 mg at 14:00). All patients with PCAF also received 0.005-0.01 mg/day of 9alpha fluorocortisol. The control group consisted of 14 healthy volunteers. Both types of replacement therapy resulted in high levels of variability in ACTH levels as compared with that in normal subjects. The areas under the curve (AUC) of the ACTH concentration over one day were not significantly different between groups 1 and 2 or between group 2 and controls. The AUC of ACTH in group 1 was significantly larger than that in controls. The mean ACTH concentration in group 1 at 07:00 and 11:00 was significantly greater than those in the other two groups. The daily rhythm of ACTH was generally closer to normal in patients given dexamethasone. Since our own clinical experience shows that at least two thirds of patients are initially given dexamethasone and that this had to be withdrawn because of the development of overdosage syndrome (weight gain, increased appetite, insomnia), it appears that there is a lack of concordance between the clinical data and the daily rhythm of ACTH secretion. When assessing the adequacy of replacement therapy in PCAF, it is important to note that the appearance of a normal rhythm of ACTH secretion over one day does not exclude the possibility of GC overdosage, with the effect that interpretation of the results of this type of measurement must take the clinical picture into account. PMID- 11430566 TI - The role of the neurodynamic factor in repair morphogenesis. PMID- 11430567 TI - Spike responses of neurons in the motor area of the cortex of elderly rabbits to specific stimuli. AB - The spike responses of neurons in the motor area of the cortex to tactile and electrocutaneous stimulation of the forelimb were studied in elderly (aged 6-7 years) rabbits. In comparison with young rabbits, the cortex of adult animals contained fewer cells responding to afferent stimulation. The activatory responses of neurons in elderly animals showed smaller increases in the spike frequency from the baseline level. Long-latency, slow activatory responses, which were not characteristic of cortical neurons in young animals, appeared; the pattern of these responses could be partially corrected by administration of acetylcholine in the vicinity of the neurons being recorded. The parameters of inhibitory responses were enzyme of the significantly different in animals of different ages. PMID- 11430568 TI - Late excitatory responses of cat motor cortex neurons to stimulation of the pyramidal tract. PMID- 11430569 TI - Impairment of learning the voluntary control of posture in patients with cortical lesions of different locations: the cortical mechanisms of pose regulation. AB - The process of learning to produce voluntary changes in the position of the center of pressures using biological feedback was studied by stabilography in patients with hemipareses due to cerebrovascular lesions in the zone supplied by the middle cerebral artery. There were significant impairments to learning in all groups of patients, who had lesions in different sites, demonstrating that cortical mechanisms are involved in learning to control posture voluntarily. These studies showed that patients with lesions in the right hemisphere had rather greater deficits in performing the task than those with lesions in the left hemisphere. There were significant differences in the initial deficit in performing the task on the first day of training depending on the side of the lesion. All groups of patients differed from healthy subjects in that significant learning occurred only at the initial stages of training (the first five days). Learning at the initial stage in patients with concomitant lesions of the parietal-temporal area and with combined lesions with motor, premotor, and parietal-temporal involvement was significantly worse and the level of task performance at the end of the initial stage was significantly worse than in patient with local lesions of the motor cortex. The level of learning was independent of the severity of the motor deficit (paresis, spasticity), but was associated with the severity of impairment of the proprioceptive sense and the severity of disruption to the upright posture (asymmetry in the distribution of support pressures, amplitude of variation in the position of the center of pressures). The learning process had positive effects on the severity of motor impairment and on the asymmetry of the distribution of support pressures in the standing posture. Reorganization of posture during bodily movements occurred mainly because of impairment to the developed "non-use" stereotype of the paralyzed lower limb. PMID- 11430570 TI - The formation of lateralized motor habits in rats. AB - Wistar rats were trained to obtain food from a narrow tube feeder using the forepaw in conditions of free choice of which limb to use. It was found that the presence or absence of limb preference, identified in short preliminary tests, often did not coincide with the nature and degree of lateralization formed during the process of prolonged training. Achievement of the maximum degree of lateralization by the animals finally forming the right-handed group was found to require longer average training periods than was the case for left-handed rats. PMID- 11430571 TI - Linked spike activity of rabbit neocortex neurons during self-stimulation of the lateral hypothalamus. AB - Cross-correlation and autocorrelation histograms were constructed to study the linkage between the spike activity of neurons in the visual and sensorimotor areas of both cerebral hemispheres in rabbits over periods of 10 sec after self stimulation of the right and left lateral hypothalamus. Self-stimulation was followed by an increase in the number of pairs of cells in which neurons in the sensorimotor area fired after cells in the visual cortex (from 24 +/- 6% to 44 +/ 6%). Analysis of the spectrum of linked discharges revealed increases in the alpha and theta frequency ranges: the main increases in the right hemisphere were in the alpha range, while the main increases in the left hemisphere were in the theta range. Interhemisphere asymmetry in the interaction of cortical neurons was not associated with the side of the hypothalamus subjected to stimulation. PMID- 11430572 TI - The effects of sodium nitrite on neocortical neuron activity during performance of defensive and inhibitory conditioned reflexes. AB - Administration of the NO-generating substance sodium nitrite to conscious non immobilized rabbits at a dose of 11 mg/kg (s.c.) decreased the intensity and duration of the short-latency "modality-specific" components of the responses of neurons in the visual cortex (to flashes of light) and sensorimotor cortex (to pain reinforcement). Decreases in neuron activation in the visual cortex in response to flashes of light occurred independently of their biological significance. i.e., as the signal for a defensive reflex and on the background of conditioned inhibition. The long-latency activatory components of the response of sensorimotor and visual cortex neurons to pain reinforcement, the inhibitory pause in the responses of visual cortex neurons to flashes of light, and the disinhibitory effect of pain reinforcement showed smaller changes after sodium nitrite. The results obtained here support the suggestion that different neuromediators are involved in transmitting "modality-specific" and "modality-non specific" influences to neocortical neurons during learning, and provide the basis for suggesting that sodium nitrite has a neurotropic action when given systemically. PMID- 11430573 TI - The role of muscarinic cholinoceptors in the retrieval of an operant food-related conditioned reflex in cats. AB - Experiments on cats showed that lesions affecting retrieval of an operant food procuring reflex. occurring on a background of systemic administration of the centrally-acting muscarinic cholinoceptor blocker scopolamine (a non-selective M cholinoceptor blocker) and trihexyphenidyl (a relatively selective M1 cholinoceptor blocker), might be associated with the central and peripheral side effects of these blockers, preventing performance of the conditioned reflex. It was established that when no side effects were present (low doses of trihexyphenidyl, 1 mg/kg), blockade of M1-cholinoceptors led to selective loss of the motor operant reflex while contextual behavior and other conditioned responses were retained or led to errors in performance of the reflex: this appears to be evidence that derangement of launching and performing the motor program is the most important component of the conditioned reflex. Systemic administration of trihexyphenidyl at a dose of 10 mg/kg, scopolamine at doses of 0.03 and 0.06 mg/kg, and the peripherally-acting non-selective blocker methylscopolamine at a dose of 0.03 mg/kg led to changes in the general functional state (disturbances in the emotional-motivational sphere), the extent of which depended on the individual sensitivity of the animal to the anticholinergic agents. The presence of side effects led to complete cessation of conditioned reflex activity, though this appeared not to be associated with memory impairment. PMID- 11430574 TI - Neuron activity in the prefrontal cortex of the brain in rats with different typological characteristics in conditions of emotional stimulation. AB - Male Wistar rats were separated according to the "emotional resonance" method (groups of animals avoiding ("altruists") and not avoiding ("egotists") the pain cries of partner rats) and neuron activity in the prefrontal areas of the cortex was studied in the right and left hemispheres. Assessments were made of changes in the frequency of nerve cell spike activity (in relation to the baseline activity of neurons in sated animals) in rats subjected to one day of food deprivation and after electrical stimulation of emotionally positive (lateral hypothalamus) and negative (tegmentum of the midbrain) brain structures and after exposure to the pain cries of partner rats. The results of these experiments revealed a series of differences in the cell activities of the two groups of rats. In conditions of hunger, the discharge frequency in the "altruists" was higher than that in "egotists." Cortical neuron responses to positive stimulation were greater than those to negative stimulation in rats of both groups. Intracerebral stimulation produced significantly greater increases in discharge frequency in neurons of both prefrontal areas of the cortex in "altruists" than in "egotists." In both groups of rats, neurons in the right hemisphere responded to emotionally negative stimulation with significantly greater activation than cells in the left hemisphere, while activity in the left hemisphere was greater in conditions of emotionally positive stimulation. "Altruists" showed significantly greater neuron responses during exposure to pain cries from "victim" rats in both the right and left hemispheres. The responses of "egotists" to "victim" cries were not significantly different from baseline activity levels. PMID- 11430575 TI - The effects of forelimb deafferentation on the post-natal development of synaptic plasticity in the hippocampus. AB - The effects of partial deafferentation of the forelimb on the development of long term potentiation in the hippocampus of rats aged 13-18 days were studied. Long term potentiation in hippocampus field CA1 was of greater amplitude and duration in control rats at 16-18 days of post-natal ontogenesis than in adult animals. Partial deafferentation by section of the median nerve in the forelimb on the 13th day of life led to the disappearance of this excess at 16-18 days. The peak in synaptic plasticity occurred later in operated animals--on day 17--and was much less marked than in controls. The decreases in the amplitude and duration of long-term potentiation in hippocampal field CA1 in operated animals provides evidence for a decrease in the sensitivity and/or number of NMDA receptors. This suggests that partial deafferentation of one limb may lead not to a decrease but to an increase in spike and synaptic activity in the hippocampus, which in normal conditions may affect the maturation of the plastic properties of synaptic transmission associated with the expression and positions of NMDA receptors. The level of long-term potentiation in sham-operated rats was significantly greater than in controls of the same age. This significant increase in NMDA-dependent long-term potentiation may be explained by a decrease in the level of activation due to anesthesia. It is suggested that the decrease in the spike activity of cells receiving signals from the median nerve may be compensated for by activation of other specific and non-specific inputs. PMID- 11430576 TI - Some physiological manifestations of the activity of the gene controlling the predisposition to pendulum-like movements in rats. AB - The physiological actions of the gene controlling the predisposition to stereotypic hyperkinesia in the form of pendulum-like movements (PM) and, probably, a form of spontaneous nystagmus in rats with albinism are manifest as a number of behavioral characteristics (total motor activity, emotionality, startle reflex intensity, sensitivity to serotonin 5-HT2 receptors as assessed in terms of the intensity of head twitching, and predisposition to cataleptic responses). A number of parameters showed differences between hybrids of gray handling tolerant rats and carriers of the PM gene on the one hand, and between gray rats and animals not carrying the PM gene on the other. Some behavioral characteristics of rats with PM were closer to those of Wistar rats than to those of rats without PM. This, as well as the high frequency of PM in Wistar rats, indicates that the gene responsible for the manifestations of PM is not pathological, but controls a variety of adaptive features of the nervous system. The interaction between PM and the predisposition to catalepsy is biphasic in nature and is described by a curve in the form of an inverted U. PMID- 11430577 TI - Dopamine-dependent inhibition of glycine release in the nucleus accumbens of the rat brain during food consumption. AB - Studies of Sprague-Dawley rats using in vivo intracerebral dialysis and high performance liquid chromatography with electrochemical detection were used to investigate glycine release into the intercellular space of the nucleus accumbens during food consumption. The results showed that food consumption led to decreases in glycine levels in the intercellular space of the nucleus accumbens. Administration of the sodium channel blocker tetrodotoxin (1 microM), but not the glutamate reuptake blocker D,L-threo-hydroxyaspartate (1 mM), prevented the food related behavior-induced decrease in glycine levels in the nucleus accumbens. Eating of food after administration of the dopamine D2 receptor blocker raclopride (10 microM) into the nucleus accumbens was accompanied by an increase in the glycine level in the intercellular space of this structure. These data provide evidence for the neural regulation of glycine release in the nucleus accumbens during food-related behavior, mediated via dopamine D2 receptors. PMID- 11430578 TI - Involvement of the rat caudate nucleus in the immunostimulatory effect of DAGO. AB - The involvement of the caudate nucleus, i.e., the terminal zone of the nigrostriatal dopaminergic system, in neuroimmunostimulation during the activation of mu opioid receptors by the highly specific agonist DAGO. Single doses of DAGO (100 microg/kg) in sham-operated control Wistar rats induced significant increases in the numbers of direct IgM-antibody-forming and total rosette-forming cells at the peak of the immune response after immunization with sheet red blood cells. The experiments showed that bilateral electrolytic lesioning of the caudate nucleus in rats suppressed the immune response, demonstrating its involvement in neuroimmunomodulation. Since the effect of immunostimulation induced by DAGO disappeared when given to animals with caudate nucleus lesions, it was concluded that this structure is involved in activatory immunogenesis via mu opioid mechanisms. PMID- 11430579 TI - Brain serotonin metabolism during water deprivation and hydration in rats. AB - The effects of two-day water deprivation and hyperhydration (provision of 4% sucrose solution for 48 h) on levels of serotonin and its major metabolite 5 hydroxyindoleacetic acid (5-HIAA) in the midbrain and hypothalamus were studied in Wistar rats. The rates of diuresis (0.05 +/- 0.01 and 0.84 +/- 0.12 ml/h/100 g in water deprivation and hyperhydration respectively) and urine osmolality (1896 +/- 182 and 50 +/- 13 mOsm/kg) reflected increases and decreases in blood vasopressin levels. Water deprivation was associated with a significant increase in 5-HIAA levels in the midbrain and hypothalamus, along with a decrease in serotonin levels and a three-fold increase in serotonin catabolism (the 5 HIAA:serotonin concentration ratio). Hyperhydration induced moderate increases in serotonin and 5-HIAA levels in the hypothalamus with no changes in the midbrain. The blood corticosterone level doubled in water deprivation and decreased in hyperhydration. It is suggested that activation of the serotoninergic system induces a complex adaptive reaction in water deprivation. including mechanisms specific for the regulation of water-electrolyte homeostasis and non-specific stress mechanisms (vasopressin and corticoliberin secretion). PMID- 11430580 TI - Neuropharmacological analysis of the role of the serotoninergic system in forming hyperalgesia in neurotic rats. PMID- 11430581 TI - Morphofunctional changes due to thyroliberin in nonapeptidergic cells in living hippocampal slices from rats. PMID- 11430582 TI - Local characteristics of the temporal organization of tissue basophils in the dura mater in adult rats. PMID- 11430583 TI - Endothelium-derived relaxing factors: a focus on endothelium-derived hyperpolarizing factor(s). AB - Endothelium-derived hyperpolarizing factor (EDHF) is defined as the non-nitric oxide (NO) and non-prostacyclin (PGI2) substance that mediates endothelium dependent hyperpolarization (EDH) of vascular smooth muscle cells (VSMC). Although both NO and PGI2 have been demonstrated to hyperpolarize VSMC by cGMP- and cAMP-dependent mechanisms, respectively, and in the case of NO by cGMP independent mechanisms, a considerable body of evidence suggests that an additional cellular mechanism must exist that mediates EDH. Despite intensive investigation, there is no agreement as to the nature of the cellular processes that mediates the non-NO/PGI2 mediated hyperpolarization. Epoxyeicosatrienoic acids (EET), an endogenous anandamide, a small increase in the extracellular concentration of K+, and electronic coupling via myoendothelial cell gap junctions have all been hypothesized as contributors to EDH. An attractive hypothesis is that EDH is mediated via both chemical and electrical transmissions, however, the contribution from chemical mediators versus electrical transmission varies in a tissue- and species-dependent manner, suggesting vessel-specific specialization. If this hypothesis proves to be correct then the potential exists for the development of vessel and organ selective vasodilators. Because endothelium-dependent vasodilatation is dysfunctional in disease states (i.e., atherosclerosis), selective vasodilators may prove to be important therapeutic agents. PMID- 11430584 TI - Antiproliferative effect of brief exposure to cholera toxin in vascular smooth muscle cells: role of cAMP and protein kinase A. AB - The effect of cholera toxin (CTX), an activator of the adenylate cyclase-coupled G protein alpha(s) subunit, was studied on cultured vascular smooth muscle cell (VSMC) proliferation. Continuous exposure (48 h) to CTX as well as 2-min pretreatment of VSMC with CTX led to the same level of cAMP production, inhibition of DNA synthesis, and arrest in the G1 phase without induction of necrosis or apoptosis in VSMC. Protein kinase A (PKA) activity in CTX-pretreated cells was transiently elevated by 3-fold after 3 h of incubation, whereas after 48 h it was reduced by 2-fold compared with baseline values without modulation of the expression of its catalytic alpha subunit. The PKA inhibitors H89 and KT 5720 did not protect VSMC from the antiproliferative effect of CTX. Two-dimensional electrophoresis was used to analyze the influence of CTX on protein phosphorylation. After 3 h of incubation of CTX-pretreated cells, we observed both newly-phosphorylated and dephosphorylated proteins (77 and 50 protein species, respectively). After 24 h of incubation, the number of phosphorylated proteins in CTX-treated cells was decreased to 39, whereas the number of dephosphorylated proteins was increased to 106. In conclusion, brief exposure to CTX leads to full-scale activation of cAMP signaling and evokes VSMC arrest in the G1 phase. PMID- 11430585 TI - Endothelium-dependent desensitization to angiotensin II in rabbit aorta: the mechanisms involved. AB - The aim of this study was to characterize the role of the endothelium in angiotensin II-desensitization and its mechanisms of action. Rabbit aortic rings were exposed to increasing doses of angiotensin II (Ang II, 10(-9) to 2.5 x 10( 6)) to generate two cumulative dose-response curves (CDRC I and II). A 50-min interval separated CDRC I and II. Desensitization was observed at all doses in unrubbed aortic tissue and at lower doses in rubbed aortic tissue. Tachyphylaxis was greater in arteries with endothelium. Treatment of intact rings with L-N(G) nitroarginine methyl ester (L-NAME, 10(-4) M) did not prevent this phenomenon. However, indomethacin (10(-5) M) and miconazol (10(-6) M) attenuated Ang II desensitization. Treatment of unrubbed rings with nifedipine (10(-6) M) and cromakalim (10(-6) M) inhibited the effect of indomethacin. To confirm the involvement of K+ channels, unrubbed and rubbed aortic rings were treated with the K(Ca2+) blockers apamin (10(-7) M), tetraethylammonium (TEA, 10(-3) M), and iberiotoxin (10(-8) M), and the K(ATP) blocker glibenclamide (10(-5) M). In both arteries apamin, TEA, and glibenclamide abolished the tachyphylaxis without changes in the maximal response. Iberiotoxin diminished Ang II-desensitization in rubbed but not unrubbed arteries. Results from this study suggest that Ang II desensitization involves endothelium-dependent and -independent mechanisms. Endothelium-dependent desensitization could be mediated by a cyclooxygenase cytochrome P450 product, which could act by increasing K(Ca2+) channel activity. PMID- 11430586 TI - Neutrophil response to prolonged exercise in immune-competent and RAG2/gamma c null mice. AB - The two aims of this study were (i) to compare the effects of prolonged exercise on circulating neutrophil number and muscle myeloperoxidase (MPO) activity between RAG2/gamma c null and immune-competent mice, and (ii) to evaluate the general suitability of the lymphocyte-deficient RAG2/gamma c null strain for use in exercise models of immune regulation. RAG2/gamma c null (male and female) and C57BL/6 (congenic immune-competent, male) mice were assigned to either control (C) or treadmill exercise (EX, 22 m/min, 90 min, 6% grade) groups. EX mice were killed immediately (EX0) or 24 h (EX24) after exercise. RAG2/gamma c null males had significantly (P < 0.05) fewer circulating CD45+ cells and higher %CD45+ neutrophils than did C57BL/6 males, independent of exercise. A significant interaction was observed for the effects of exercise and gender on %CD45+ neutrophils in the blood. At EX24, gastrocnemius (Gastroc) MPO significantly increased in EX mice. Gastroc MPO activity was 44% and 35% higher in RAG2/gamma c null vs. C57BL/6 males, and in female vs. male RAG2/gamma c null mice, respectively. Heart MPO activity did not differ between strains or among treatments. We concluded that the Rag2/gamma c null strain is a suitable model for future investigations on immune regulation following acute exercise stress. PMID- 11430587 TI - A1 receptor activation decreases fatigue in mammalian slow-twitch skeletal muscle in vitro. AB - To test the hypothesis that adenosine improves skeletal muscle cell function, we exposed curarized mouse soleus and extensor digitorum longus (EDL) to a range of concentrations of adenosine (10(-9) M to 10(-5) M). Muscles contracted in Krebs Henseleit bicarbonate buffer (27 degrees C, 95% O2 and 5% CO2) for 500 ms at 50 Hz once every 90 s. Soleus fatigued significantly less with adenosine present at concentrations of 10(-8) M and higher than with the Krebs-Henseleit vehicle control. Adenosine significantly improved force generation or delayed fatigue of EDL only with the initial adenosine challenge. To investigate the receptor population involved, we exposed soleus to agonists specific for A1 receptors (N6 cyclopentyladenosine, CPA), or A2 receptors (CGS 21680 hydrochloride, CGS), or A3 receptors (N6-benzyl-5'-N-ethylcarboxamidoadenosine, BNECA). CPA (A1) significantly decreased fatigue compared with the Krebs-Henseleit vehicle control at concentrations of 10(-9) M and higher. Muscles exposed to the A2 and A3 agonists did not differ from a Krebs-Henseleit plus methanol control. Phenylephrine (10(-6) M), an alpha-adrenergic agonist that increases the concentration of inositol triphosphate (IP3), significantly improved developed force in soleus. Neither a permeable cAMP analog, 8-bromo-cAMP (10(-5) M), nor a beta, agonist, isoproterenol (10(-6) M), had an effect on force generation in the soleus when compared with a saline control. Thus adenosine slowed fatigue in slow twitch skeletal muscle through A1 receptors. PMID- 11430588 TI - Calcium homeostasis in a clonal pituitary cell line of mouse corticotropes. AB - Calcium homeostasis was studied following a depolarization-induced transient increase in [Ca2+]i in single cells of the clonal pituitary cell line of corticotropes, AtT-20 cells. The KCl-induced increase in [Ca2+]i was blocked in (i) extracellular calcium-deficient solutions, (ii) external cobalt (2.0 mM), (iii) cadmium (200 microM), and (iv) nifedipine (2.0 microM). The mean increase in [Ca2+]i in single cells in the presence of an uncoupler of mitochondrial function [carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone, FCCP, 1 microM] was 54 +/- 13 nM (n = 9). The increase in [Ca2+]i produced by FCCP was greater either during or following a KCl-induced [Ca2+]i load. However, FCCP did not significantly alter the clearance of calcium during a KCl-induced rise in [Ca2+]i. Fifty percent of the cells responded to caffeine (10 mM) with an increase in [Ca2+]i (191 +/- 24 nM; n = 21) above resting levels; this effect was blocked by ryanodine (10 microM). Thapsigargin (2 microM) and 2,5 di(-t-butyl) 1,4 hydroquinone (BuBHQ, 10 microM) produced increases in [Ca2+]i (47 +/- 11 nM, n = 6 and 22 +/- 4 nM, n = 8, respectively) that increased cell excitability. These results support a role for mitochondria and sarco-endoplasmic reticulum calcium stores in cytosolic [Ca2+]i regulation; however, none of these organelles are primarily responsible for the return of [Ca2+]i to resting levels following this KCl-induced [Ca2+]i load. PMID- 11430589 TI - Contractile function in vitro of slow-twitch skeletal muscle from weanling mice subjected to wasting malnutrition. AB - Our hypothesis was that malnutrition sufficient to produce weight loss in weanling mice would decrease the ability of slow-twitch skeletal muscle to develop and maintain force. We isolated muscles from 3 groups (n = 5) of weanling C57BL/6J mice of both sexes (i) mice at 19 days of age serving as zero-time or baseline controls (CONT) (ii) mice fed for the next 14 days with a low-protein diet that produces features of incipient kwashiorkor (LPD) and (iii) mice fed for the next 14 days with a complete diet (NORM). Muscles were also obtained from 5 adult mice 7-9 months of age (MAT). We stimulated the soleus at 50 Hz for 500 ms at 0.6 tetanic contractions per min (tet x min(-1)), 6 tet x min(-1), and 30 tet x min(-1) in Krebs-Henseleit bicarbonate buffer at 27 degrees C gassed with 95% O2 and 5% CO2. The initial developed force (mN x mm(-2)) at 0.6 tet x min(-1) did not differ across groups (CONT 211.7 +/- 16.0, LPD 274.2 +/- 41.6, NORM 246.8 +/- 38.0, MAT 210.8 +/- 10.6). The fatigue rate (mN x mm(-2) x min(-1)) at 6 tet x min(-1) was significantly slower in muscles from CONT (0.6 +/- 0.3) and LPD (0.6 +/- 0.4) than in NORM (2.4 +/- 0.6) and MAT (2.3 +/- 0.2). At 30 tet x min(-1), the fatigue rate (mN x mm(-2) x min(-1)) did not differ across groups (CONT 2.4 +/- 0.5, LPD 2.7 +/- 0.5, NORM 2.5 +/- 0.4, MAT 2.0 +/- 0.2). After stimulation at 6 tet x min(-1) and 30 tet x min(-1), only muscles from CONT and LPD recovered to 100%. Because muscles from LPD mice developed equal force, fatigued less, and recovered from fatigue to a greater extent than muscles from NORM mice, we rejected the hypothesis. The function of the tissue remaining in the muscles from LPD mice approximated that of muscles from mice at 19 days of age rather than muscles from either mice of the same age fed a complete diet or adult mice. PMID- 11430591 TI - Diminished molecular response to doxorubicin and loss of cardioprotective effect of dexrazoxane in Egr-1 deficient female mice. AB - Doxorubicin (DOX) and VP16 are DNA topoisomerase II inhibitors yet only DOX induces an irreversible cardiotoxicity, likely through DOX-induced oxidative stress. Egr-1 is overexpressed after many stimuli that increase oxidative stress in vitro and after DOX-injection into adult mice in vivo. To investigate Egr-1 function in the heart, we compared the molecular and histological responses of wild type (+/+) and Egr-1 deficient (-/-) female mice to saline, DOX, VP16, the cardioprotectant dexrazoxane (DZR), or DOX+DZR injection. DOX, and to a lesser extent VP16, induced characteristic increases in cardiac muscle and non-muscle genes typical of cardiac damage in +/+ mice, whereas only beta-MHC and Sp1 were increased in -/- mice. DZR-alone treated +/+ mice showed increased cardiomyocyte transnuclear width without a change to the heart to body weight (HW/BW) ratio. However, DZR-alone treated -/- mice had an increased HW/BW, increased cardiomyocyte transnuclear width, and gene expression changes similar to DOX injected +/+ mice. DZR pre-injection alleviated DOX-induced gene changes in +/+ mice; in DZR+DOX injected -/- mice the increases in cardiac and non-muscle gene expression were equal to, or exceeded that, detected after DOX-alone or DZR-alone injections. We conclude that Egr-1 is required for DOX-induced molecular changes and for DZR-mediated cardioprotection. PMID- 11430590 TI - Oxidations of 17beta-estradiol and estrone and their interconversions catalyzed by liver, mammary gland and mammary tumor after acute and chronic treatment of rats with indole-3-carbinol or beta-naphthoflavone. AB - Altered cytochrome P450-catalyzed metabolism of 17beta-estradiol (E2) and estrone (E1) in the liver and (or) extrahepatic tissues may affect estrogen-sensitive tumorigenesis. We examined the effects of oral treatments of (i) indole-3 carbinol (13C) at 250 or 500 mg/kg or beta-naphthoflavone (beta-NF) at 40 mg/kg of body weight (bw)/day from 51 to 54 days of age (acute regimen), and (ii) 13C at 250 mg/kg or beta-NF at 20 mg/kg bw given 3x/week from 10 to 22 weeks of age (chronic regimen) in female Sprague-Dawley rats. We determined the effects of these treatments on the P450 content and P450 (CYP)-specific activities in the liver, P450-dependent metabolism of E2 and E1 by the liver and mammary gland, and interconversion of E1 and E2 catalyzed by 17beta-hydroxysteroid dehydrogenase (17beta-HSD) in these tissues and malignant mammary tumors. 13C at the two levels of acute regimen elicited similar responses. Acute and chronic treatments with 13C, but not beta-NF, increased P450 content approximately 2-fold. 13C, and to a lesser extent beta-NF, increased CYP1A1 and CYP1A2 probe activities in liver up to 117- and 27- fold, respectively, and after acute regimens, that of CYP3A by approximately 1.8-fold. 13C also increased activity of CYP2B up to 100-fold. Overall hepatic metabolism of E2 and E1, which was approximately 2-fold greater at 55 than 155 days of age, was increased (approximately 2.8-fold) by 13C with 2 , 4-, 16alpha-, 6alpha-, 6beta-, and 15alpha-hydroxy (OH) comprising > or = 54, 3, 2, approximately 2, approximately 5, 7, and 2%, respectively, of E1 and E2 metabolites. Acute regimens of beta-NF increased 2- and 15alpha-OH-E2 (62 and 5% of total) from E2 and 2-, 4-, and 6alpha-OH-E1 + 6beta-OH-E1 (32, 13, and 4% of total) from E1. Mammary gland metabolized E2 to E1 and small amounts of 15alpha-, 4-, 16alpha-, 6beta-, and 6alpha-OH-E2. After the acute IC3 regimen, E2 was also converted to 2-OH-E2. 17Beta-HSD-catalyzed oxidation of E2 was favored in the liver and reduction of E1 was favored in mammary gland and tumor (= 1% of hepatic activity). An increased (approximately 2-fold) ratio of reductive to oxidative activities in malignant mammary tumors by chronic 13C regimen may stimulate tumor growth. This is the first report showing that after chronic oral regimens, the 13C-, but not beta-NF-, induced changes in CYP complement led to elevated E2 and E1 metabolism. The persistent effects of increased putative carcinogenic and estrogenic 4- and 16alpha-OH as well as 6alpha- and 6beta-OH-E2 and 6beta-OH-E1 might counteract those of the less estrogenic 2-OH metabolites, thus accounting for the lack of suppression of mammary tumorigenesis by 13C in our previous study. PMID- 11430592 TI - Effect of progesterone on the contractile response of isolated pulmonary artery in rabbits. AB - The purpose of this study was to assess the direct effect of progesterone on rabbit pulmonary arteries and to examine the mechanism of its action. Rings of pulmonary artery from male rabbits were suspended in organ baths containing Krebs solution, and isometric tension was measured. The response to progesterone was investigated in arterial rings contracted with noradrenaline (NA), KCl, and CaCl2. The effects of endothelium, nitric oxide (NO), prostaglandins, cyclic GMP (cGMP), and the adrenergic beta-receptor on progesterone-induced relaxation were also assessed. Progesterone inhibited the vasocontractivity to NA, KCl, and CaCl2, and relaxed rabbit pulmonary artery. The relaxing response of progesterone in pulmonary artery was significantly reduced by removal of endothelium, inhibitors of nitric oxide synthase and guanylate cyclase, but not by prostaglandin synthase inhibitor and blockage of the adrenergic beta-receptor. In Ca2+-free (0.1 mM EGTA) Krebs solution, progesterone inhibited NA-induced contraction that was intracellular Ca2+-dependent, but didn't affect the contraction of extracellular Ca2+-dependent component. Our results suggest that progesterone induces relaxation of isolated rabbit pulmonary arteries partially via NO and cGMP. Progesterone may also inhibit Ca2+ influx through potential dependent calcium channels (PDCs) and Ca2+ release from intracellular stores. PMID- 11430593 TI - A decade of progress in allogeneic hematopoietic cell transplantation: 1990-2000. PMID- 11430594 TI - Human herpesvirus-8 and Kaposi's sarcoma: relationship with the multistep concept of tumorigenesis. AB - Kaposi's sarcoma (KS) develops through discrete inflammatory-angiogenic stages of polyclonal nature (early-stage lesions) to monomorphic nodules of spindle-shaped cells that can be clonal (late-stage lesions) and resemble true sarcomas. Molecular and epidemiological studies indicate that development of KS is tightly associated with infection by the human herpesvirus-8 (HHV-8). However, only individuals with specific conditions of immunodysregulation develop KS. In these individuals the systemic and tissue increase of Th-1-type cytokines (IC) reactivate HHV-8 infection, leading to increased viral load, antibody titers, and an expanded cell tropism that precedes the clinical appearance of KS. Recruitment of the virus into tissues by infected monocytes and other cell types is facilitated by the endothelial cell activation due to IC. In clinical lesions, HHV-8 infection increases with lesion stage and in late-stage lesions most of the spindle cells are latently infected, whereas only few lyrically infected cells are present, suggesting that latent genes may have a role in the transformation of the early inflammatory-hyperplastic lesion into a real sarcoma. The development of tumors, however, is regulated through a multistep process based on the acquisition by cells of several different capabilities leading to malignant growth. Here we review the available data on the expression of HHV-8-encoded genes in primary KS lesions and, in view of their biological activity, analyze their potential function in different steps of tumorigenesis. By this pragmatic approach interesting insights into potential key functions of HHV-8-encoded genes are found and steps of potential cooperativity with other viral factors (HIV-1 Tat) in the pathogenesis of KS are identified. PMID- 11430595 TI - Reactivation and role of HHV-8 in Kaposi's sarcoma initiation. AB - Kaposi's sarcoma (KS) is an angioproliferative disease occurring in several clinical-epidemio-logic forms but all associated with infection by the human herpesvirus-8 (HHV-8). At least in early stages, KS is a reactive disease associated with a state of immune dysregulation characterized by CD8+ T-cell activation and production of Th1-type inflammatory cytokines (IC) that precedes lesion development. In fact, evidence indicates that IC can trigger lesion formation by inducing the activation of endothelial cells that leads to adhesion and tissue extravasation of lymphomonocytes, spindle cell formation, and angiogenesis, and HHV-8 reactivation that, in turn, leads to virus spread to all circulating cell types and virus dissemination into tissues. Due to virus escape mechanisms and deficient immune responses toward HHV-8, virus reactivation and spread are not controlled by the immune system but induce immune responses that may paradoxically exacerbate the reactive process. The virus is recruited into "activated" tissue sites where it finds an optimal environment for growth. In fact, viral load is very low in early lesions, whereas almost all spindle cells are infected in late-stage lesions. Although early KS is a reactive process of polyclonal nature that can regress, in time and in the presence of immunodeficiency, it can progress to a true sarcoma. This is likely due to the long-lasting expression of HHV-8 latency genes in spindle cells associated with the deregulated expression of oncogenes and oncosuppressor genes and, for AIDS KS, with the effects of the HIV-1 Tat protein. PMID- 11430596 TI - A role for secondary V(D)J recombination in oncogenic chromosomal translocations? AB - Chromosomal translocations are hallmarks of certain lymphoproliferative disorders. Indeed, in many leukemias and lymphomas, translocations are the transforming event that brings about malignancy. Recurrence of the immunoglobulin (Ig) and T-cell receptor (Tcr) loci at the breakpoints of oncogenic chromosomal translocations has led to speculation that the lymphocyte-specific process of V(D)J rearrangement, which is necessary for the generation of functional Ig and TCR antigen receptors on B and T lymphocytes, mediates translocation. Recent studies have led to a fuller understanding of the molecular mechanisms of V(D)J rearrangement and have revealed that the V(D)J recombinase possesses latent transposase activity. These studies have led to plausible models of illegitimate V(D)J recombination producing chromosomal translocations consistent with those present in lymphomas and leukemias. Errors of V(D)J recombination may even generate lymphomas with the phenotypes of mature cells. For example, follicular and Burkitt's lymphomas have been classified by phenotype and somatic genotype as malignant germinal center (GC) B or post-GC B cells. The GC is a site of affinity maturation where B cells undergo V(D)J hypermutation and Ig class switch; in addition, much evidence has accumulated to suggest that GC B cells may also support secondary V(D)J recombination. Interestingly, all three of these elements, genomic plasticity, mutation, and translocation breakpoints near switch sites or recombinational elements, are characteristic of certain lymphomas. The high frequency of lymphomas carrying these GC markers suggests that the GC reaction may play a significant role in lymphomagenesis. PMID- 11430597 TI - Recombinant immunotoxins in targeted cancer cell therapy. AB - Targeted cancer therapy in general and immunotherapy in particular combines rational drug design with the progress in understanding cancer biology. This approach takes advantage of our recent knowledge of the mechanisms by which normal cells are transformed into cancer cells, thus using the special properties of cancer cells to device novel therapeutic strategies. Recombinant immunotoxins are excellent examples of such processes, combining the knowledge of antigen expression by cancer cells with the enormous developments in recombinant DNA technology and antibody engineering. Recombinant immunotoxins are composed of a very potent protein toxin fused to a targeting moiety such as a recombinant antibody fragment or growth factor. These molecules bind to surface antigens specific for cancer cells and kill the target cells by catalytic inhibition of protein synthesis. Recombinant immunotoxins are developed for solid tumors and hematological malignancies and have been characterized intensively for their biological activity in vitro on cultured tumor cell lines as well as in vivo in animal models of human tumor xenografts. The excellent in vitro and in vivo activities of recombinant immunotoxins have lead to their preclinical development and to the initiation of clinical trail protocols. Recent trail results have demonstrated potent clinical efficacy in patients with malignant diseases that are refractory to traditional modalities of cancer treatment: surgery, radiation therapy, and chemotherapy. The results demonstrate that such strategies can be developed into a separate modality of cancer treatment with the basic rationale of specifically targeting cancer cells on the basis of their unique surface markers. Efforts are now being made to improve the current molecules and to develop new agents with better clinical efficacy. This can be achieved by development of novel targeting moieties with improved specificity that will reduce toxicity to normal tissues. In this review, the design, construction, characterization, and applications of recombinant immunotoxins are described. Results of recent clinical trails are presented, and future directions for development of recombinant immunotoxins as a new modality for cancer treatment are discussed. PMID- 11430598 TI - Nurses' behavioural intentions towards self-poisoning patients: a theory of reasoned action, comparison of attitudes and subjective norms as predictive variables. AB - BACKGROUND: The incidence of self-poisoning is on the increase. Most patients who self-poison are dealt with initially in the general hospital. Therefore, the type and quality of care self-poisoning patients receive will depend, in part, on how they are viewed by nursing staff within the general hospital setting. A knowledge and understanding of the attitudes held by nurses towards self-poisoning patients is therefore important to those involved in the planning and delivery of care towards this client group. Previous studies have examined health care professionals' attitudes towards people who self-poison. Usually, however, these have not focused specifically on nurses' attitudes, and they have ignored the relationship between the attitudes expressed by staff and their intentions to engage in subsequent caring behaviour of one sort or another. It is hence unclear how the findings of such studies are relevant or applicable to nursing policy and practice. AIMS: The present study aims to address these limitations using a methodology informed by the theory of reasoned action. The study aims to separate out the distinctive roles played by nurses' own attitudes, and the social pressures represented by other people's attitudes, in determining the types of caring behaviour in which nurses intend to engage when dealing with self poisoning patients. DESIGN/METHODS: The study adopts a questionnaire-based approach incorporating two specially designed vignettes. RESULTS: The results show that nurses' own attitudes, and what they believe about the attitudes of others, predict their behavioural intentions towards self-poisoning patients. The study also shows that nurses with a more positive orientation towards self poisoning patients differ in behavioural and normative beliefs from nurses who have a less positive orientation. CONCLUSIONS: The implications for future attempts to explore the relationship between nurses' attitudes and subsequent caring behaviour are considered, along with implications for nursing policy and practice. PMID- 11430599 TI - Improving community mental health nurse targeting of people with severe and enduring mental illness: experiences from one English health district. AB - BACKGROUND: Successive governments have urged mental health service providers to target their attentions on people with severe and enduring mental illness (SEMI). However, community mental health teams (CMHTs) in general, and community mental health nurses (CMHNs) in particular, have been criticized for failing to meet this requirement. This paper reports selected findings from a wider study that assessed the impact of an initiative designed to facilitate service targeting: the establishment of registers of patients with SEMI in general practices throughout an English health district. The paper describes changes in the nature of community mental health nursing contacts with a sample of patients on these registers. METHODS: Six general practices were randomly selected from the 65 practices in the district and comparisons made between patients on the six mental health registers who either had, or did not have, community mental health nursing contact. These comparisons related to the year before the establishment of the registers, the year during which they were being established and the year following this. RESULTS: A total of 274 patients were included on the sample registers, with practices varying considerably in relation to proportions of mental health registered patients with community mental health nursing contact. Overall, the number of patients in contact with CMHNs was found to have decreased over time, except for those on level 2 of the Care Programme Approach. CONCLUSIONS: No evidence was found to support the hypothesis that the establishment of the registers had improved CMHN targeting of patients with SEMI. However, findings were inconclusive because shortages of CMHNs and the disruption associated with widespread service reorganization meant the registers were never fully implemented in practices during the study period. PMID- 11430600 TI - Different effort constructs and effort-reward imbalance: effects on employee well being in ancillary health care workers. AB - AIMS OF THE STUDY: The present study investigates the relationship between Effort Reward Imbalance (ERI) and employee well-being, using three different concepts of efforts (i.e. psychological demands, physical demands and emotional demands). BACKGROUND: The ERI model had been used as a theoretical framework, indicating that work stress is related to high efforts (i.e. job demands) and low occupational rewards (e.g. money, esteem and security/career opportunities). The ERI model also predicts that, in overcommitted workers, effects of ERI on employee well-being are stronger compared with their less committed counterparts. METHODS: A cross-sectional survey among 167 ancillary health care workers of two nursing homes was conducted. Multiple univariate logistic regression analyses were used to test the relationship between ERI and employee well-being. RESULTS: Results of the logistic regression analyses showed that employees with both high (psychological, physical and emotional) efforts and low rewards had higher risks of psychosomatic health complaints, physical health symptoms and job dissatisfaction (odds ratios (ORs) ranged from 5.09 to 18.55). Moreover, employees who reported both high efforts and high rewards had elevated risks of physical symptoms and exhaustion (ORs ranged from 6.17 to 9.39). No support was found for the hypothesis on the moderating effect of overcommitment. CONCLUSION: Results show some support for the ERI model; ancillary health care workers with high effort/low reward imbalance had elevated risks of poor employee well-being. In addition, results show that the combination of high efforts and high rewards is important for employee well-being. Finally, some practical implications are discussed to combat work stress in health care work. PMID- 11430601 TI - Experiencing a nurse identity: the meaning of identity to Swedish registered nurses 2 years after graduation. AB - RATIONALE: The professional identity and experiences of nurses have been focused upon in different studies AIM: This is a longitudinal study whose aim was to understand how nurses experience the meaning of their identity as nurses, when they are students and nurses 2 years after graduation. DESIGN: Data were collected through interviews once a year during education and two years after graduation, and were analysed using a phenomenological hermeneutic method, inspired by the philosophy of Paul Ricoeur. FINDINGS: The analyses of the narratives resulted in four perspectives: 'Having the patient in focus', 'Being a team leader', 'Preceptorship' and 'Task orientation'. The nurses did not change perspectives but the perspective showed a transition over time. CONCLUSION: The nurses' not changing perspective over time is understood as being a life paradigm, remaining throughout the years. PMID- 11430602 TI - Psychosocial rehabilitation after disfiguring injury or disease: investigating the training needs of specialist nurses. AB - AIM OF THIS STUDY: To investigate the training needs of specialist nurses working with patients who have disfiguring conditions. BACKGROUND: As the psychosocial impact of disfiguring illness and disease becomes more widely recognized and understood, there are increasing issues about who can provide this kind of input. Whilst nurses have been identified as the health professionals who are often the most appropriate to fulfil this role, there is less evidence that they feel confident in their skills to provide it. The volume of requests from health professionals, particularly nurses, to a lay-led organization specializing in facial disfigurement suggested both an acknowledgement of the importance of providing psychosocial support to this population, together with a perceived lack of skill in this area. DESIGN/METHOD, STUDY 1: This study used a cross-section design. Two groups of nurses working either in burns or in head and neck cancer were surveyed by questionnaire. RESULTS, STUDY 1: Both the burns group and head and neck group rated their skills in the social rehabilitation of their patient group as significantly lower than their skills in physical rehabilitation. DESIGN/METHOD, STUDY 2: Using a multiple baseline design, a pilot intervention consisting of a specifically focused resource pack and 1-day training course was delivered to eight head and neck cancer nurses. RESULTS, STUDY 2: Postintervention, only the targeted skills (those concerned with social rehabilitation) were rated as significantly improved. Research continues to establish the extent to which these skills are implemented by nurses in the clinical setting and passed on to patients. CONCLUSIONS: These findings suggest that although nurses do not feel as skilled in delivering psychosocial support to patients as they do in delivering more traditional aspects of physical nursing care, they can very easily take on this role when given simple training and access to appropriate resources. This training can be provided by a lay-led organization. PMID- 11430603 TI - Developing critical reflection for professional practice through problem-based learning. AB - AIMS: To explore the influence of current learning traditions in nursing on the development of reflection and critical reflection as professional practice skills and to offer suggestions for nursing education that will specifically facilitate the development of critical reflection. ORGANIZATIONAL CONSTRUCTS: Mezirow's transformative learning theory, Barrows conceptualization of problem-based learning (PBL). METHODS: Integrative literature review of published literature related to nursing, health science education and professional education from 1983 2000. FINDINGS: Professional education scholars concur that specialized knowledge is clearly essential for professional practice, however, they also suggest that self-consciousness (reflection) and continual self-critique (critical reflection) are crucial to continued competence. While strategies to facilitate reflection have been outlined in the literature, specific strategies to facilitate the development of critical reflection and implications for nursing education are much less clear. Advocates of reflective and critically reflective practice suggest that the development of these abilities should be inextricably linked to professional development and can be developed through active repeated guided practice. In health care, PBL based on constructivism, has been identified as one way to facilitate the development of these skills. CONCLUSIONS: Nursing learners exposed to PBL develop the ability to be reflective and critically reflective in their learning and acquire the knowledge and skill within the discipline of nursing by encountering key professional practice situations as the stimulus and focus of their classroom learning. The learners' ability to be both reflective and critically reflective in their learning is developed by critical questioning of the faculty tutor during situational analysis, learning need determination, application of knowledge, critique of resources and personal problem-solving processes, and summarization of what was learned. PMID- 11430604 TI - Meanings of good nursing care among Thai female last-year undergraduate nursing students. AB - AIMS OF THE STUDY: To describe and express the meanings of good nursing care among female last-year undergraduate nursing students of a nursing college in Bangkok, Thailand. BACKGROUND: In Thailand today, nursing educational programmes aim at producing graduates who can explore options in nursing science. METHOD: Twenty nursing students were interviewed and observed using the ethnographic method. FINDINGS: Six categories emerged, viz., compassion (give care from heart; have sympathy; be kind and helpful; have and share feelings; be friendly and be concerned; be honest), competency (have knowledge, skill and experience; be interested to learn; have responsibility; be professional; be confident; know what to do without hesitation), comfort (assist, help and treat patient as self; attend, give patient dignity and respect; have goals and give nursing care corresponding to the patients' needs), communication (have time to listen, talk and explain; be willing to listen and inform patients; meet patients' needs), creation (create new knowledge; apply theory in practice; continue MSc or PhD study; do research) and courage (advocate for patient's needs and rights, intervene for and with patient; make decisions about nursing care; face and solve patient's problems). CONCLUSIONS: The findings should be useful in the development and continuous improvement of nursing educational programmes, including curricula and teaching methods. PMID- 11430605 TI - Facilitating and obstructing factors for development of learning in clinical practice: a student perspective. AB - AIM OF THE STUDY: The aim of this study was to provide information on what the student nurses found facilitating and obstructing for their learning during clinical practice. BACKGROUND: Earlier studies of experiences of learning in clinical practice have shown that factors as the possibilities of variations of experiences, the culture of the workplace, and communication between the educational institution and health care facilities are of importance. Less is known about the opportunities which students are given in order to practise the skills that they will be expected to perform as new graduate nurses. METHODS: The experiences of 47 degree student nurses from two colleges in Sweden were gathered in weekly diaries during their final period of clinical practice. A content analysis technique was used to analyse their diaries. RESULTS: The students emphasized responsibility and independence, opportunities to practise different tasks, and receiving feedback as facilitating factors. Other perceived promoting factors included perceptions of control of the situation and understanding of the 'total picture'. Examples of obstructing factors were the nurses as supervisors not relying on the students, supervision that lacked continuity and lack of opportunities to practise. Perception of their own insufficiency and low self reliance were drawbacks for some students. CONCLUSIONS: Recommended proposals are presented to lecturers and supervising staff concerning organizational and educational changes, and changes of attitudes for elucidating the students' experiences of different facilitating and obstructing factors. Changes may contribute to making easier the students' transition into the nursing profession. PMID- 11430606 TI - Achieving consensus about pain content for child branch curricula. AB - AIM: The aim of the study was to identify nurse educators' and nurse managers' perceptions of the educational needs of children's nurses in relation to pain management. RATIONALE: Children continue to suffer unnecessary pain during hospitalization. This has been attributed, in part, to inadequate education among paediatric nurses with a number of studies demonstrating that nursing curricula need re-evaluating with respect to the content relating to pain management. In order to equip nurses to manage pain within clinical practice nurse education needs to provide them with the knowledge and skills required. It is apparent that nurse education is currently failing students in this regard. In order to evaluate nursing curricula with respect to pain management it is necessary for there to be a degree of consensus, among paediatric nurses, regarding appropriate content. METHODOLOGY: A two-round Delphi study was used to ascertain the appropriate pain content for pre-registration child branch courses. When undertaking a Delphi study a number of 'experts' in the field are identified, who complete a series of questionnaires. The first round is used to generate ideas, which are reconsidered in subsequent rounds (Hicks 1999, Reid 1988). Following analysis of the responses to each questionnaire anonymous feedback is sent to the respondents together with the next questionnaire (Walker 1994). Participants were either senior nurse educators or senior nurse managers. RESULTS: Positive correlations were found within the two groups as well as between the educators' and managers' rankings for the 14 categories identified. CONCLUSIONS: The results of this study suggest that nurse educators and nurse managers have similar views about the pain management knowledge they would expect a newly qualified child branch nurse to have. The list of appropriate pain content derived from the results of this study provides a useful tool with which to evaluate the content of child branch curricula. PMID- 11430607 TI - The needs of children visiting on adult intensive care units: a review of the literature and recommendations for practice. AB - AIM: The purpose of this paper is to review the current literature and research available and to identify specific, nursing interventions to meet the needs of child visiting within the ICU setting. BACKGROUND: According to recent surveys children are still restricted from visiting their critically ill family and friends on many adult intensive care units (ICUs) within the United Kingdom (UK). Imposing restrictive visiting policies does not respect the rights of patients and their families to be together and to support each other during a period of stress and crisis. METHOD: The motivation to undertake the study was derived from a critical incident involving a small boy who was not allowed to visit his critically ill mother. She subsequently died. Reflection on the available literature identified the value and role of intuition in expert clinical judgement, but the need to support this with evidence based knowledge. CONCLUSION: The implications for practice are discussed and recommendations for further research are made. PMID- 11430608 TI - Living with post-traumatic stress disorder: the wives'/female partners' perspective. AB - AIM OF THE STUDY: This phenomenological study examined what it was like for the wives/female partners to live with a Vietnam veteran who suffers from post traumatic stress disorder (PTSD). PROCEDURE: Audio-taped interviews were conducted with 10 women and data were examined from three overlapping phases of the veteran/partner relationship: the early phase, the middle phase and the later phase. FINDINGS: The early phase was conceptualized as a period of adjustment in which three themes were identified: (1) attractors, (2) feelings and (3) communication. The middle phase, one of enmeshment, was characterized by six themes: (1) dealing with veteran PTSD symptoms, (2) substance abuse, (3) physical and/or emotional abuse, (4) roles, (5) feelings and (6) coping techniques. Three themes depicted the later phase of resolution/healing: (1) stress related symptoms, (2) staying or leaving and (3) activities that promoted an ongoing process of resolution/healing. CONCLUSIONS: The central meaning statement that best described the experience of wives/female partners who live with a Vietnam veteran with PTSD is that the experience is a gradual process of becoming enmeshed in the veteran's pathology, with all energies being directed at minimizing the effect on self and family, culminating in intermittent movement towards resolution/healing. This study has implications for practitioners who treat Vietnam PTSD veterans and their wives or female partners. PMID- 11430609 TI - The changing training needs of clinical nurse managers: exploring issues for continuing professional development. AB - AIMS OF THE STUDY: To identify areas where clinical nurse managers perceived that they would benefit from further training and to make recommendations for planning future programmes to meet their needs. BACKGROUND: The effectiveness of the clinical nurse manager has traditionally been associated with maintaining standards of care. Continuing professional development (CPD) is essential to ensure this important group feel adequately prepared to perform their role and has been recognized as an important factor in maintaining job satisfaction and reducing wasteful staff turnover. A review of the literature indicated that since the 1980s the CPD needs of clinical nurse managers have tended to be overlooked despite increasing complexity of the tasks expected of them. Thus it appeared that a fresh study to address these needs would be justified and should take into account sources of work-related stress and variables relating to job satisfaction. METHODS: The study involved clinical nurse managers employed in all four acute hospital National Health Service (NHS) trusts where training needs were served by a major inner city educational consortium. Data collection proceeded in two stages. Initially interviews were undertaken with a random sample of 15 clinical nurse managers to provide in-depth, qualitative data. This information was used to develop a survey questionnaire distributed to the remaining 182 clinical nurse managers in each of the trusts. RESULTS: Data from the interviews indicated that clinical nurse managers appeared to feel clinically competent but generally experienced lack of confidence when dealing with a range of issues, in particular; human resources, managing budgets, deputizing for senior colleagues across the trust ('acting up') and using information technology in everyday practice. Response rate to the survey was good (65%). The results corroborated the interview findings, indicating a need for updating in the same wide range of topics. There were few differences in training needs across all four trusts. Clinical nurse managers who perceived that they had been well prepared for their role demonstrated higher levels of job satisfaction. Perceived preparation for clinical leadership was the same irrespective of the employing trust, length of time in post and other sociodemographic variables. CONCLUSIONS: The study findings confirm that CPD remains a major issue for clinical nurse managers in the United Kingdom (UK) and that providing opportunities for such development may be an important factor in enhancing job satisfaction. The study findings should help those providing CPD to plan more effectively for this group and have implications for staff recruitment and retention. PMID- 11430610 TI - Initiation factors for substance abuse. AB - AIM: The study reported in this paper investigated factors that led to the initial use and abuse of drugs in 237 addicts receiving treatment at the Psychological Medicine Hospital, Kuwait. METHOD: A self-administered questionnaire was designed to elicit information regarding socio-demographic data and initiation patterns. Questions about initiation patterns were related to personal, familial and social variables in addition to accessibility to drugs. RESULTS: Almost all the sample were male (99.2%) and 42.6% were married. The findings were grouped into (1) factors associated with drug abuse and (2) initial/ significant factors for drug abuse. One way ANOVA showed that there was a significant statistical association between current age and source of drug. Also, there was a significant statistical association between the age of the addict when first started on drugs, and the source of drugs. Multiple regression analysis was also computed for initial significant factors. One factor 'excessive unsupervised free time' was statistically significant. RECOMMENDATIONS: Strategies for primary, secondary and tertiary interventions in addition to recommendation for further studies are suggested. PMID- 11430611 TI - Behaviour modification and gentle teaching workshops: management of children with learning disabilities exhibiting challenging behaviour and implications for learning disability nursing. AB - BACKGROUND: Challenging behaviours (behaviour difficulties) represent a problem of considerable clinical significance for learning disability nurses, and a source of much human distress. Gentle teaching is a relatively new approach to dealing with behavioural difficulties, and has been received with enthusiasm by clinicians, but has so far received little empirical support. The current study attempted to compare gentle teaching with a well-established alternative (behaviour modification) and a control group. OBJECTIVES: To examine the comparative effectiveness of gentle teaching, behaviour modification and control interventions for challenging behaviour amongst children with learning disabilities. DESIGN: Nonrandomized controlled trial. SETTING: Service users' homes in East Yorkshire. PARTICIPANTS: Seventy-seven children who presented with learning disabilities and challenging behaviour (behaviour difficulties) and their parents. PROCEDURE: One-day workshops in were offered by recognized authorities in either behaviour modification or gentle teaching that were not otherwise involved with the research project. Forty-one participants were recruited to the gentle teaching condition; 36 to behaviour modification; 26 to the control group. Random allocation was not possible, because of the slow uptake by interested parents. Measures was preintervention, and at assessment points up until 12 months following intervention. ANALYSIS: Quantitative analysis of pre post differences between the groups, using t-test. RESULTS: In general, no significant differences were found between the treatment groups and controls. Significant improvements were found for both gentle teachingand behaviour modification children over controls on the AAMR ABS XVII (social engagement) subscale. Controls had more contact with medical practitioner (GP) services than behaviour modification children and less than gentle teaching children. CONCLUSION: Although very few differences were found between the three groups, those that did exist generally favoured behaviour modification. Implications for service provision and learning disability nursing practice are described. PMID- 11430612 TI - The meaning of suicidal psychiatric inpatients' experiences of being treated by physicians. AB - Twenty-one psychiatric inpatients who had thought about, expressed a wish to, or even attempted to commit suicide narrated their experiences of receiving treatment from physicians. The interview texts were transcribed and interpreted using a phenomenological-hermeneutic method inspired by the philosophy of Ricoeur. Two main themes and eight themes were found. Participating Approach: being with the patients, listening to patients without prejudice, trusting each other, respecting patients' integrity; and Observing Approach: leaving patients to themselves, listening to patients with prejudice, mistrusting each other, not respecting patients' integrity. These findings were interpreted in the light of the writings of Hegel, Buber and Cissna and Sieburg. The suicidal patients expressed their need for confirmation in their interaction with physicians, that is they needed to meet with respect and interest when staying in the hospital with suicidal thoughts or after suicidal attempts. PMID- 11430613 TI - Mechanisms of action in the treatment of anxiety. AB - Anxiety and depressive disorders share many features, suggesting a common set of physiologic substrates. Recent research has indicated that mood can be categorized into 3 components by factor analysis: (1) somatic anxiety (a factor relatively specific to panic disorder), (2) anhedonic depression (which includes symptoms related to motivation and enjoyment and found to be specific to depression), and (3) general distress (a factor that cuts across all depressive and anxiety disorders studied). Antidepressant drugs, particularly serotonin reuptake inhibitors and serotonin receptor modulators, are effective for a wide variety of anxiety and depressive disorders. The impact on both anxiety and depression may be a result of an effect on a common set of physiologic targets relevant to the general distress dimension. At a cellular level, the antidepressants target components of the stress-adaptation system in brain, which may explain these common effects. On the other hand, there appear to be differences in the relative impact of serotonergic and noradrenergic drugs on the spectrum of distress and motivational symptoms. Basic research and clinical research suggest that serotonergic agents may be preferentially effective for symptoms of general distress, whereas catecholaminergic agents may target anhedonic depression symptoms. PMID- 11430614 TI - Serotonergic and noradrenergic reuptake inhibitors: prediction of clinical effects from in vitro potencies. AB - This article reviews the pharmacology of antidepressants, particularly focusing on those that act acutely by blocking the reuptake of norepinephrine (NE) and/or serotonin (5-HT). Such drugs have a very wide range of potencies, measured in vitro, to inhibit the reuptake of these biogenic amines. As a group, the selective serotonin reuptake inhibitors (SSRIs) are the most potent at inhibiting the reuptake of 5-HT. Some tricyclic antidepressants (TCAs), such as desipramine and nortriptyline, are much more potent at blocking NE reuptake than 5-HT reuptake, as is the new non-TCA drug reboxetine. Among SSRIs, paroxetine is most potent at blocking the reuptake of NE. When considering whether such potencies measured in vitro translate into pharmacologic effects clinically, it is necessary to know how much drug gets to its site of therapeutic action, presumably the brain. Most, but not all, antidepressants are extensively bound to plasma proteins, and this binding limits considerably the penetration of these drugs across the blood-brain barrier. The amount of drug present in the extracellular fluid (ECF) of brain approximates the non-protein-bound drug concentration in plasma. Comparison of the concentration of antidepressants in ECF with their potencies to inhibit the reuptake of 5-HT and/or NE reveals why some drugs block the reuptake of these biogenic amines in either a selective or nonselective manner. This analysis reveals that venlafaxine may be unique among antidepressants in having a dose-dependent nonselectivity; at low doses it acts primarily as an SSRI, but at higher doses it inhibits the reuptake of NE as well. PMID- 11430615 TI - Comorbidity, neurobiology, and pharmacotherapy of social anxiety disorder. AB - Social anxiety disorder is a common psychiatric illness that imposes persistent functional impairment and disability on persons who have the disorder. The disorder is characterized by a marked and persistent fear of social or performance situations in which embarrassment may occur. It is the most prevalent of any anxiety disorder and is the third most common psychiatric disorder after depression and alcohol abuse. Social anxiety disorder typically begins during childhood with a mean age at onset between 14 and 16 years and is sometimes preceded by a history of social inhibition or shyness. Persons who have social anxiety disorder either endure or avoid social situations altogether because the fear of embarrassment causes such intense anxiety; such avoidance may ultimately interfere with occupational and/or social functioning and lead to significant disability. The duration of social anxiety disorder is frequently lifelong, and there is a high degree of comorbidity with other psychiatric disorders. Social anxiety disorder is a serious illness that frequently runs a chronic course and is associated with significant morbidity. Patients should be treated aggressively using pharmacotherapeutic agents that can be tolerated over the long term. Cognitive-behavioral therapy should also be considered in treatment planning. Efforts to increase the recognition of social anxiety disorder as a common, distressing, and disabling condition are critical. This article discusses the comorbidity, neurobiology, and pharmacotherapy of social anxiety disorder. PMID- 11430616 TI - Contemporary management of comorbid anxiety and depression in geriatric patients. AB - Anxiety and depression in elderly people are major public health problems in the United States. Recognition and treatment of these conditions will likely gain more attention in the next 30 to 50 years because of the projected growth of the geriatric population. As in many younger patients, the most common presentation of anxiety in elderly patients is comorbid anxiety and depression. Although age is not a risk factor for either anxiety or depression, factors associated with aging--such as increased medical burdens and loss of independence--are substantial risk factors for development of these conditions. Moreover, there is a close association in older people between untreated mental illness and exacerbation of physical illness. Some of the newer antidepressants are more appropriate long-term options for the treatment of comorbid anxiety and depression than either benzodiazepines or tricyclic antidepressants. The newer antidepressants can decrease symptoms, improve quality of life, and potentially promote healthier outcomes in geriatric patients who have comorbid anxiety and depression and/or comorbid mental and physical illness. PMID- 11430617 TI - Treatment of anxiety disorders to remission. AB - Treating anxiety disorders to remission should be the goal of all practitioners. A remitted patient should be well, both in symptoms and function, and be indistinguishable from a never-ill counterpart. The definition of remission in patients with anxiety disorders should also be clear, practical, and easy to use. It is useful to measure response in an objective way, such as with standardized instruments appropriate for the disorder, and to develop remission criteria specific to each disorder. This article proposes remission criteria, using standardized measures, for 5 common anxiety disorders: panic disorder, social anxiety disorder, generalized anxiety disorder, obsessive-compulsive disorder, and post-traumatic stress disorder. PMID- 11430618 TI - The effect of under and over refractive correction on visual performance and spectacle lens acceptance. AB - As a follow-on from a previous study by Miller, Kris and Griffiths (1997, Optom. Vis. Sci. 84, 521-526), we investigated the effect of small prescription errors on spatial visual performance and spectacle lens acceptability. We included both negative and positive errors and binocular and monocular errors. Data were collected on 15 young adult subjects. Clinical measures were taken of pupil size, ocular dominance, binocular visual acuity, negative and positive relative accommodation, distance and near heterophorias, and stereopsis. Visual performance was measured with the best correction and for +/-0.50 D spherical binocular and monocular errors. Subjects wore spectacles, each containing a different error in turn, for 2 days and compared them with a reference pair. Following the wearing period subjects assessed the performance of the spectacles by completing a short questionnaire. The only ocular tests for which these small prescription errors had significant effects were binocular visual acuity and negative relative accommodation. No significant relationship was observed between any of the clinical test results and overall lens acceptance. Despite this, the reference pair was generally perceived as better than the test pairs containing errors, and a considerable proportion of subjects reported pairs with errors as being unacceptable. In conclusion, the questionnaire findings support the need for both accurate monocular and binocular refractions. Subjects differed in their criteria for judging lens acceptability. PMID- 11430619 TI - Economic costs of cataract surgery using a rigid and a foldable intraocular lens. AB - Optimal delivery of healthcare requires consideration of various costs. A foldable intraocular lens (IOL) is more expensive than an equivalent rigid IOL. However, surgical and post-operative costs may make a foldable IOL economically preferable. We compared the economic costs of cataract surgery plus implantation of a foldable IOL with implantation of a rigid IOL. Prospective audit of the clinical records of 82 pseudophakes; 39 implanted with a rigid IOL and 43 implanted with a foldable IOL by one surgeon. Average follow-up periods were 25 +/- 7 months and 23 +/- 5 months respectively. There was no difference between the two groups for the follow-up period (P = 0.55), number of post-operative complications (P = 0.25) or cost of post-operative visits (P = 0.83). The cost of single-use theatre equipment was greater for the rigid-IOL group (P= 0.0001). The total identified cost per patient was greater for the foldable-IOL group (P = 0.0001). Despite possible technical advantages, implantation of the foldable IOL did not provide an economic benefit, either in the initial cost or in the costs of post-operative care. Over the 2-year period, implanting with the rigid IOL cost, on average, Pound Sterling57 less per patient. Despite this economic difference, a cost-benefit analysis is required, since other factors may be more important. PMID- 11430620 TI - Detection of uveal melanoma by optometrists in the United Kingdom. AB - In the United Kingdom, most uveal melanomas are detected by optometrists. Prospectively collected data on patients with uveal melanoma presenting to optometrists were analysed retrospectively to determine: (a) the proportion of patients who were asymptomatic at the time of tumour detection, (b) the proportion of symptomatic patients reporting that their tumour was missed at their initial presentation and (c) the clinical features related to immediate tumour detection. The 223 patients had a mean age of 59.7 years, a mean tumour diameter of 11.3 mm and a mean tumour thickness of 4.6 mm. Symptoms were present in 122 patients (55%) and were associated with large tumour size (p < 0.0001) and male gender (p = 0.003), with more males tending to have a large tumour (p = 0.004). Seventy-nine percent of symptomatic patients reported that their tumour was detected at their first visit. Failure of tumour detection in symptomatic patients was associated with absence of tumour extension posterior to equator (p < 0.0001). PMID- 11430621 TI - Optimal reading speed in simulated cataract: development of a potential vision test. AB - Previous studies suggest that optimal reading speed is unaffected by cataract, yet is significantly reduced in age-related macular disease. This raises the question of whether a reading speed test could be developed to assess potential vision after cataract surgery. In this study, 20 young subjects, with and without a simulation of dense cataract, read aloud Bailey-Lovie word charts. From the results, critical print size, optimal reading speed and word acuity were calculated. The simulated cataract reduced word acuity and increased the critical print size, yet optimal reading speed remained unchanged. Measurements were also taken with the charts in reversed contrast polarity. Reversing the contrast polarity of the charts improved the word acuity and optimal reading speed with the cataract simulation. The results suggest that optimal reading speed could be used as a potential vision test. Further investigations using patients with cataract and ARMD are required. PMID- 11430622 TI - Residual visual functions of guide dog owners in the UK. AB - Numbers of guide dog owners (GDOs) in the United Kingdom reached 4700 by the end of 1998. Despite this growing trend, little is known about the nature of their visual loss. This paper reports the results of a national three-centre investigation into the residual visual functions and ophthalmic conditions of guide dog owners. Random samples of GDOs (Scotland n = 82, England n = 77, and Northern Ireland n = 87) underwent a detailed visual analysis and interview. GDOs had an overall median age of 53 years. Nationally, they make up just 2.4% of the registered blind population. All GDOs were found to have profound loss of visual acuity, contrast sensitivity or visual fields, but only 43% were totally blind. GDOs in Scotland retained higher levels of residual visual function than those in the other two regions. The main causes of visual loss were congenital and early onset degenerative eye disease (retinitis pigmentosa 18%, optic atrophy 10%). Results taken in conjunction with epidemiological registration trends suggest that the past growth in numbers of GDOs is unlikely to be sustainable. Implications for mobility service providers are discussed. It is suggested that increased optometric input and a multidisciplinary approach could assist present and potential guide dog owners. PMID- 11430623 TI - Corneal surface area: an index of anterior segment growth. AB - Corneal surface area and perimeter were assessed as novel indices to monitor anterior segment growth, using chicks reared under different photoperiods. We obtained central and mid-peripheral corneal curvatures using photokeratometry. Anatomical tracings of the anterior corneal surface also were made from freeze dried non-fixed preparations of the anterior segments of the same eyes. Using either photokeratometry or anatomical data, the profile of the anterior corneal surface was fit to a general equation for conical sections; corneal surface area was estimated from surfaces of revolution. Optical techniques modeled the chick cornea as a circle or as an ellipse closely resembling a circle. The anatomical technique, in contrast, modeled the chick corneal profile as a hyperbola. Potential explanations of this discrepancy are discussed. Regardless of which model is evaluated, the corneal surface area and perimeter of two-week-old chicks are affected by the photoperiod of rearing. Corneal surface area in particular proved more sensitive than conventional measurements in identifying anterior segment effects of rearing under different photoperiods. Analysis of corneal area may prove useful in evaluating the mechanisms governing anterior segment growth. PMID- 11430624 TI - Optical and biometric relationships of the isolated pig crystalline lens. AB - AIM: To investigate the interrelationships between optical and biometric properties of the porcine crystalline lens, to compare these findings with similar relationships found for the human lens and to attempt to fit this data to a geometric model of the optical and biometric properties of the pig lens. METHODS: Weight, focal length, spherical aberration, surface curvatures, thickness and diameters of 20 isolated pig lenses were measured and equivalent refractive index was calculated. These parameters were compared and used to geometrically model the pig lens. RESULTS: Linear relationships were identified between many of the lens biometric and optical properties. The existence of these relationships allowed a simple geometrical model of the pig lens to be calculated which offers predictions of the optical properties. CONCLUSIONS: The linear relationships found and the agreement observed between measured and modeled results suggest that the pig lens confirms to a predictable, preset developmental pattern and that the optical and biometric properties are predictably interrelated. PMID- 11430625 TI - The effect of conventional CR39 and Fresnel prisms on high and low contrast acuity. AB - The effect of conventional CR39 and Fresnel prisms on high and low contrast letter acuity was studied. Visual acuity of the fully corrected better eyes of 15 subjects was measured with the high (90%) and low (10%) contrast logMAR letter charts, while they wore prisms of varying power. The results showed that when the power of the conventional CR39 prism and the Fresnel prism reached 10 prism dioptres and 5 prism dioptres, respectively, significant reduction (1 tail t test, p < 0.05) of the high and low contrast acuity occurred. The Fresnel prism caused a significantly greater acuity reduction than the conventional CR39 prism for powers ranging from 5 to 30 prism dioptres for both contrasts. The rate of acuity reduction with increasing prism power was greater with the low contrast targets than with the high contrast targets for both prisms. In addition, the rate of acuity reduction with increasing prism power was greater with the Fresnel prism than with the conventional CR39 prism for both contrasts. The conventional CR39 prism reduced acuity by a ratio of about 0.8-0.9 of that of the Fresnel prism for powers ranging from 5 to 20 prism dioptres and by about 0.7 for 30 prism dioptres. These ratios applied for both high and low contrast acuity, and therefore were independent of the level of contrast used. PMID- 11430626 TI - The gradient index and spherical aberration of the lens of the human eye. AB - We present equations for analysing the primary spherical aberration of the crystalline lens, both as a whole and of its contributing parts. We apply these equations to two examples of gradient index lenses. The spherical aberration of one lens is well outside that expected for real lenses, but that of the other is within this range. We assess the sensitivity of the aberration to changes in surface shape and gradient index distribution. The process is applicable to other models of the gradient index of the lens. PMID- 11430627 TI - The stenopaeic slit: an analytical expression to quantify its optical effects in front of an astigmatic eye. AB - The stenopaeic slit is a trial case accessory used in subjective refraction, especially when high astigmatism is present. In spite of its simplicity, the effect of the slit when it is not oriented along one of the principal meridians of the examined eye is difficult to predict, even in terms of classical geometrical optics. In this paper, the optical principles of the slit are considered with full details in the theoretical framework of the dioptric power space. An analytical expression to obtain the residual refractive error when a stenopaeic slit is placed in front of an astigmatic eye at any orientation is deduced. In the light of these results, some aspects of the clinical procedure are discussed. PMID- 11430628 TI - The Pulsair 3000 tonometer--how many readings need to be taken to ensure accuracy of the average? AB - Manufacturers of non-contact tonometers recommend that a number of readings are taken on each eye, and an average obtained. With the Keeler Pulsair 3000 it is advised to take four readings, and average these. This report analyses readings in 100 subjects, and compares the first reading, and the averages of the first two and first three readings with the "machine standard" of the average of four readings. It is found that, in the subject group investigated, the average of three readings is not different from the average of four in 95% of individuals, with equivalence defined as +/- 1.0 mmHg. PMID- 11430629 TI - Series on "lifestyle drugs". PMID- 11430630 TI - Anti-obesity drugs: use, mechanisms and clinical importance. AB - The high prevalence of obesity (>20% in industrialized countries) and its serious health risks, cause severe individual and public costs. Pharmacological interventions are available, such as dietary products and drugs acting on the absorption of nutrients, the production of hormones regulating the feeling of hunger and satiety, and the expenditure of energy. The individual benefit of these pharmacological interventions depends on the efficiency of these measures and the side-effects associated with them. The afflicted patient, and very often even the treating physician, are unable to evaluate the risk/benefit ratio of these drugs for the individual patient. This article describes the possibilities and limitations of pharmacological interventions to treat obesity and gives information on the indications for dietary, pharmacological and surgical interventions. PMID- 11430631 TI - Alpha-interferon (IFN-alpha) treatment of chronic hepatitis C: analysis of some predictive factors for the response. AB - Interferon (IFN)-alpha is recognized today as the treatment of choice for chronic hepatitis virus C-related disease. Several factors are able to influence the response to the therapy. In the last few years, several types of IFN-alpha have been used with varying therapeutic results. In order to define the role played by the type of IFN-alpha, as a predictive factor of response to therapy, we studied clinical and biological features of 128 patients (80 males and 48 females, mean age 58.5+/-9.7 years) divided into four homogeneous groups. Four types of IFN alpha were administered at the same dose and for an identical period. Lymphoblastoid was administered to Group A; recombinant 2a to Group B; recombinant 2b to Group C and leukocyte to Group D. The results showed that the best response was achieved using natural IFN. Statistical evaluation of the predictive factors was carried out using bivariate and multivariate analysis. IFN alpha was associated with different response rates for viral and biochemical remission at the end of the treatment and at the end of 12 months' follow-up period. Therefore, the type of IFN administered may influence the response to therapy and important independent single predictive factors of response should be taken into account. The type of IFN used may determine how well the disease is controlled. PMID- 11430632 TI - Metabolic functions of the liver during chemotherapy in children with acute lymphoblastic leukemia. AB - OBJECTIVE: The purpose of the present work was estimation of liver function using the phenazone test and commonly used biochemical tests in children with acute lymphoblastic leukemia (ALL) during anticancer treatment. METHODS: Observations were carried out in the same 21 patients with ALL before the beginning of chemotherapy, after Protocol I and after Protocol M of the antileukemic treatment carried out according to the program BFM 86. RESULTS: The applied chemotherapy inhibited phenazone elimination. Both phenazone half-life and metabolic clearance rate were significantly different in patients after treatment with anticancer drugs, especially with high-dose of methotrexate (MTX), from those in patients before the beginning of chemotherapy (p < 0.001). Moreover, after MTX administration transaminases activity and serum bilirubin concentration were significantly higher than before treatment (p < 0.05). CONCLUSION: Our results showed that in children with acute lymphoblastic leukemia, anticancer chemotherapy decreased liver metabolic capacity. Particularly, high-dose methotrexate treatment altered the elimination of phenazone by inhibiting the activity of hepatic mixed function oxidase system. This change may lead to an increase in toxicity of active drugs which are metabolized by this enzyme system. In addition, altered activity of liver metabolic function can impair transformation of prodrugs to active forms. It should be considered in selection of individual drug dosages. The objective estimation of the type and degree of liver dysfunction can only be achieved by the combination of a quantitative phenazone dynamic test and static biochemical tests. PMID- 11430633 TI - Pharmacoutilization of antihypertensive drugs: a model of analysis. AB - OBJECTIVES: To identify clinical and economic indicators of pharmacoutilization of antihypertensive drugs. PATIENTS AND METHODS: 4614 subjects receiving a first prescription for amlodipine, atenolol, fosinopril, indapamide, or losartan were included in the study. All prescriptions filled during the study period from January 1, 1997 to December 31, 1998 were considered. A retrospective analysis was carried out on information recorded in the drug database. The percentage of patients continuing, discontinuing, and switching the initial treatment, duration of treatment, and doses used were calculated together with total costs. RESULTS: A large proportion of patients (65.1%) discontinued the treatment. From the analysis of the mean daily dose taken by patients who continued the treatment, it was found that many subjects took a drug dosage which was below the therapeutic dose range, whereas the administration of doses above the therapeutic range occurred only occasionally. Continuation of treatment accounted for 48.1% of total costs, switching accounted for 20.8%, and discontinuation represented 31.1% of total expenditures. CONCLUSIONS: With adequate markers, helpful data can be collected for monitoring the quality of antihypertensive drug prescriptions and the rational usage of resources in the general practice setting. PMID- 11430634 TI - Dose linearity and steady state pharmacokinetics of the new antiparkinson agent budipine after oral administration. AB - The dose-dependency of budipine pharmacokinetic characteristics was studied. Eighteen healthy male subjects were given 10, 20 and 30 mg oral single doses according to a randomized, open, 3-period crossover design. Additionally, the steady state conditions were investigated after repeated intake of 10 mg t.i.d for 10 days and compared to the 10 mg single dose. The area under the concentration vs time curve (AUC) and the maximum serum concentration (Cmax) showed a linear increase in line with ascending doses of orally given budipine. Time to maximum serum concentration (tmax) and terminal half-life (t1/2) were independent of the administered dose. As compared to the 10 mg single dose pharmacokinetics, the repeated oral administration of budipine 10 mg t.i.d. resulted in an increase in AUC of 11% and 93% for budipine and its metabolite p OH-budipine, respectively. In clinical practice, a predictable response in proportion to the dose is to be expected. PMID- 11430635 TI - Indomethacin activates carbonic anhydrase and antagonizes the effect of the specific carbonic anhydrase inhibitor acetazolamide, by a direct mechanism of action. AB - OBJECTIVES: In this paper we investigated the effect of indomethacin, acetazolamide and their combination in vitro and in vivo on carbonic anhydrase (CA) isozymes. METHOD: In vitro experiments followed the effect of the two substances at concentrations between 10(-8)-10(-4) M on purified human red cell CA I and II as well as on human gastric mucosa CA IV using dose-response relationships. Kinetic studies were also performed. The effects of single and combined administration of indomethacin and acetazolamide on red cell CA and on gastric acid secretion were studied in vivo. RESULTS: Indomethacin, in vitro and in vivo. induces an increase in erythorcyte CA I and CA II activity. Acetazolamide, a specific inhibitor of CA, reduces the activity of CA I and CA II from red cells. Indomethacin completely antagonizes CA activity, i.e. abolishes the inhibitory effect of acetazolamide on CA. In humans, an increase or decrease in erythrocyte CA II activity is correlated with an increase or decrease in gastric acid secretion. CONCLUSIONS: Our results show that indomethacin, a known cyclooxygenase (COX) inhibitor, is also an activator of CA. Our data also prove that indomethacin is not only an activator of CA but also antagonizes the effect of acetazolamide, a specific inhibitor of this enzyme. In view of the role of CA in acid-base balance as well as the fact that an increase or decrease in its activity is accompanied by an increase or decrease in intra- and extracellular pH, our results suggest that: firstly, CA activation induced by indomethacin might cause changes in COX activity; secondly, PGs are synthetized as a consequence of the changes in COX activity, a hypothesis that requires further study. PMID- 11430636 TI - Relative bioavailability of imipramine (Tofranil) coated tablets in healthy volunteers. AB - OBJECTIVES: Imipramine is a tricyclic antidepressant drug with a considerable hepatic first-pass metabolism resulting in highly variable pharmacokinetic characteristics and desipramine as active major metabolite. This study describes the bioavailability of 3 formulations of imipramine. METHODS: In a randomized, three-period crossover study, 18 healthy male Caucasian subjects received single oral doses of Tofranil 25, Tofranil mite (10 mg) and an aqueous solution containing 25 mg imipramine-HCl. Serum concentrations of imipramine-HCl and its main metabolite desipramine were measured. The pharmacokinetic characteristics, Cmax, AUC, t1/2 and tmax were determined and the relative bioavailability of the two coated tablet formulations was calculated with the aqueous solution as reference. Safety and tolerability were assessed using vital signs, ECG, clinical laboratory and adverse event recording. RESULTS: The relative bioavailabilities of Tofranil 25 and Tofranil mite were 97% and 81%, respectively. The study medication was well tolerated. CONCLUSIONS: A sufficiently high extent of absorption was found for the test formulations ensuring therapeutic efficacy. PMID- 11430637 TI - Direct and indirect sexual selection and quantitative genetics of male traits in guppies (Poecilia reticulata). AB - The ornamentation and displays on which sexual attractiveness and thus mating success are based may be complex and comprise several traits. Predicting the outcome of sexual selection on such complex phenotypes requires an understanding of both the direct operation of selection on each trait and the indirect consequences of selection operating directly on genetically correlated traits. Here we report the results of a quantitative genetic analysis of the ornamentation, sexual attractiveness, and mating success of male guppies (Poecilia reticulata). We analyze male ornamentation both from the point of view of single ornamental traits (e.g., the area of each color) and of composite measures of the way the entire pattern is likely to be perceived by females (e.g., the mean and contrast in chroma). We demonstrate that there is substantial additive genetic variation in almost all measures of male ornamentation and that much of this variation may be Y linked. Attractiveness and mating success are positively correlated at the phenotypic and genetic level. Orange area and chroma, the area of a male's tail, and the color contrast of his pattern overall are positively correlated with attractiveness and/or mating success at the phenotypic and genetic levels. Using attractiveness and mating success as measures of fitness, we estimate gradients of linear directional sexual selection operating on each male trait and use equations of multivariate evolutionary change to predict the response of male ornamentation to this sexual selection. From these analyses, we predict that indirect selection may have important effects on the evolution of male guppy color patterns. PMID- 11430638 TI - Landscape structure and hierarchical genetic diversity in the brook charr, Salvelinus fontinalis. AB - Explaining the extent, causes, and consequences of biotic distributions in space is fundamental to our understanding of how species evolve and cope with particular environments. Yet, identifying extrinsic barriers to migration imposed by landscape structure and predicting their impacts on intraspecific genetic diversity remains a major challenge in population biology. In this study, 30 populations (771 individuals) of brook charr (Salvelinus fontinalis, Salmonidae) representing six major river drainages from Maine, USA, were characterized at six microsatellite loci to quantify the role of landscape features, such as habitat size, altitude, contemporary and historical connectivity. in shaping genetic diversity at three spatial scales: within lakes, within river drainages, and among river drainages. Within-population expected heterozygosity was negatively correlated with altitude, whereas no significant correlation was observed with lake size. Conversely, the extent of heterozygote deficiency within lakes was negatively associated with habitat size. The hierarchical analysis of genetic variance revealed that the extent of among-drainage differentiation was unexpectedly low relative to the pronounced population structuring within drainage. Geographically proximate St. John and Penobscot River drainages were characterized by opposite effects of altitude and geographic distance in shaping the pattern of population differentiation within drainages. The geographic pattern of differentiation among drainages could not be accounted for either by an isolation by distance or by a stepwise range expansion model. Overall, this study provided evidence for the role of contemporary landscape features in shaping the observed pattern of genetic diversity at smaller geographic scales (within and among populations within river drainage). On a broader geographic scale, contemporary landscape structure appeared to be only a minor factor determining the observed pattern of genetic structuring among drainages. These results add to the increasing evidence for nonequilibrium conditions between drift and migration in a wide array of animal taxa. The development of more realistic theoretical descriptions of nonequilibrium population structure thus appears to be important to better understand the relative influence of historical and ecological factors in shaping genetic variation in young habitats, such as recently deglaciated areas. PMID- 11430639 TI - Phylogeography of the trumpetfishes (Aulostomus): ring species complex on a global scale. AB - The distribution of circumtropical marine species is limited by continental boundaries, cold temperate conditions, and oceanic expanses, but some of these barriers are permeable over evolutionary time scales. Sister taxa that evolved in separate ocean basins can come back into contact, and the consequences of this renewed sympatry may be a key to understanding evolutionary processes in marine organisms. The circumtropical trumpetfishes (Aulostomus) include a West Atlantic species (A. maculatus), an Indian-Pacific species (A. chinensis), and an East Atlantic species (A. strigosus) that may be the product of a recent invasion from the Indian Ocean. To resolve patterns of divergence and speciation, we surveyed 480 bp of mitochondrial DNA cytochrome b in 196 individuals from 16 locations. Based on a conventional molecular clock of 2% sequence divergence per million years, the deepest partitions in a neighbor-joining tree (d = 0.063-0.082) are consistent with separation of West Atlantic and Indian-Pacific species by the Isthmus of Panama, 3-4 million years ago. By the same criteria, trumpetfish in the East Atlantic were isolated from the Indian Ocean about 2.5 million years ago (d = 0.044-0.054), coincident with the advent of glacial cycles and cold-water upwelling around South Africa. Continental barriers between tropical oceans have only rarely been surmounted by trumpetfishes, but oceanic barriers do not appear to be substantial, as indicated by weak population partitioning (phiST = 0.093) in A. chinensis across the Indian and Pacific Oceans. Finally, morphological and mitochondrial DNA data indicate hybridization of A. strigosus and A. maculatus in Brazil. After 3-4 million years and a globe-spanning series of vicariant and dispersal events, trumpetfish lineages have come back into contact in the southwest Atlantic and appear to be merging. This ring species phenomenon may occur in a broad array of marine organisms, with clear implications for the production and maintenance of biodiversity in marine ecosystems. PMID- 11430640 TI - Speed and stamina trade-off in lacertid lizards. AB - Morphological and physiological considerations suggest that sprinting ability and endurance capacity put conflicting demands on the design of an animal's locomotor apparatus and therefore cannot be maximized simultaneously. To test this hypothesis, we correlated size-corrected maximal sprint speed and stamina of 12 species of lacertid lizards. Phylogenetically independent contrasts of sprint speed and stamina showed a significant negative relationship, giving support to the idea of an evolutionary trade-off between the two performance measures. To test the hypothesis that the trade-off is mediated by a conflict in morphological requirements, we correlated both performance traits with snout-vent length, size corrected estimates of body mass and limb length, and relative hindlimb length (the residuals of the relationship between hind- and forelimb length). Fast running species had hindlimbs that were long compared to their forelimbs. None of the other size or shape variables showed a significant relationship with speed or endurance. We conclude that the evolution of sprint capacity may be constrained by the need for endurance capacity and vice versa, but the design conflict underlying this trade-off has yet to be identified. PMID- 11430641 TI - Cultural inheritance as a mechanism for population sex-ratio bias in reptiles. AB - Although natural populations of most species exhibit a 1:1 sex ratio, biased sex ratios are known to be associated with non-Mendelian inheritance, as in sex linked meiotic drive and cytoplasmic inheritance (Charnov 1982; Hurst 1993). We show how cultural inheritance, another type of non-Mendelian inheritance, can favor skewed primary sex ratios and propose that it may explain the female-biased sex ratios commonly observed in reptiles with environmental sex determination (ESD). Like cytoplasmic elements, cultural traits can be inherited through one sex. This, in turn, favors skewing the primary sex allocation in favor of the transmitting sex. Female nest-site philopatry is a sex-specific, culturally inherited trait in many reptiles with ESD and highly female-biased sex ratios. We propose that the association of nest-site selection with ESD facilitates the maternal manipulation of offspring sex ratios toward females. PMID- 11430642 TI - Host life history and the evolution of parasite virulence. AB - We present a general epidemiological model of host-parasite interactions that includes various forms of superinfection. We use this model to study the effects of different host life-history traits on the evolution of parasite virulence. In particular, we analyze the effects of natural host death rate on the evolutionarily stable parasite virulence. We show that, contrary to classical predictions, an increase in the natural host death rate may select for lower parasite virulence if some form of superinfection occurs. This result is in agreement with the experimental results and the verbal argument presented by Ebert and Mangin (1997). This experiment is discussed in the light of the present model. We also point out the importance of superinfections for the effect of nonspecific immunity on the evolution of virulence. In a broader perspective, this model demonstrates that the occurrence of multiple infections may qualitatively alter classical predictions concerning the effects of various host life-history traits on the evolution of parasite virulence. PMID- 11430643 TI - Chill-coma tolerance, a major climatic adaptation among Drosophila species. AB - Most drosophilid species can be classified either as temperate or tropical. Adults of species were submitted to a cold treatment (0 degrees C) and then brought back to ambient temperature. They generally exhibited a chill coma and the time needed to recover was measured. We found in a set of 26 temperate species that recovery was rapid (average 1.8 min, range 0.15-4.9). In contrast, a long recovery time (average 56 min, range 24-120) was observed for 48 tropical species. A few species, like Drosophila melanogaster, are cosmopolitan and can proliferate under temperate and tropical climates. In 9 of 10 such species, slight genetic differences were found: a shorter recovery in temperate than in tropical populations. Comparing physiological data to phylogeny suggests that chill-coma tolerance has been a recurrent adaptation that is selected for in cold climates but tends to disappear under a permanently warm environment. This major climatic adaptation, evidenced in drosophilids, seems to occur in other insect groups also. PMID- 11430644 TI - The interspecific origin of B chromosomes: experimental evidence. AB - A centric fragment was generated during the introgression of a chromosome region from Nasonia giraulti into N. vitripennis. This neo B chromosome carries the N. giraulti or 123+ gene for wild-type eye color. Using this phenotypic effect, the transmission of this chromosome was analyzed. The supernumerary chromosome showed less than Mendelian segregation rate in meiosis and some mitotic instability manifested as mosaic phenotype for eye color. However, transmission rate and mitotic stability increased over successive generations. The transmission rate through male gametogenesis was nearly 100%. These results support the interspecific hybridization model for B chromosome origin and reveal that problems in chromosome stability can persist for several generations after "foreign chromosomes" are introduced into a different species. We suggest that hybrid zones should be investigated as possible sites for neo-B chromosome generation. PMID- 11430645 TI - Quantifying passive and driven large-scale evolutionary trends. AB - I introduce a new statistical method, analysis of skewness, for quantifying large scale evolutionary trends as a combination of both passive and driven trends. My approach is based on the skewness of subclades within a parent clade. I partition the total skewness of the parent clade into three components: (1) skewness between subclades; (2) skewness within subclades; and (3) skewness due to changes in variance among subclades. The third component corresponds to a new type of passive trend, in which overall skewness of a parent clade is due to greater variability in subclades to the right of the mean. Using this partitioning, I decompose an observed trend into two components: a driven portion and a passive portion. thus quantifying the effect of small-scale dynamics on large-scale behavior of clades. Applications are given to Miocene-Pliocene rodent size and Ordovician brachiopod muscle geometry. PMID- 11430646 TI - Predicting nuclear gene coalescence from mitochondrial data: the three-times rule. AB - Coalescence theory predicts when genetic drift at nuclear loci will result in fixation of sequence differences to produce monophyletic gene trees. However, the theory is difficult to apply to particular taxa because it hinges on genetically effective population size, which is generally unknown. Neutral theory also predicts that evolution of monophyly will be four times slower in nuclear than in mitochondrial genes primarily because genetic drift is slower at nuclear loci. Variation in mitochondrial DNA (mtDNA) within and between species has been studied extensively, but can these mtDNA data be used to predict coalescence in nuclear loci? Comparison of neutral theories of coalescence of mitochondrial and nuclear loci suggests a simple rule of thumb. The "three-times rule" states that, on average, most nuclear loci will be monophyletic when the branch length leading to the mtDNA sequences of a species is three times longer than the average mtDNA sequence diversity observed within that species. A test using mitochondrial and nuclear intron data from seven species of whales and dolphins suggests general agreement with predictions of the three-times rule. We define the coalescence ratio as the mitochondrial branch length for a species divided by intraspecific mtDNA diversity. We show that species with high coalescence ratios show nuclear monophyly, whereas species with low ratios have polyphyletic nuclear gene trees. As expected, species with intermediate coalescence ratios show a variety of patterns. Especially at very high or low coalescence ratios, the three-times rule predicts nuclear gene patterns that can help detect the action of selection. The three-times rule may be useful as an empirical benchmark for evaluating evolutionary processes occurring at multiple loci. PMID- 11430647 TI - Parasites and the evolution of self-fertilization. AB - Assuming all else is equal, an allele for selfing should spread when rare in an outcrossing population and rapidly reach fixation. Such an allele will not spread, however, if self-fertilization results in inbreeding depression so severe that the fitness of selfed offspring is less that half that of outcrossed offspring. Here we consider an ecological force that may also counter the spread of a selfing allele: coevolution with parasites. Computer simulations were conducted for four different genetic models governing the details of infection. Within each of these models, we varied both the level of selfing in the parasite and the level of male-gamete discounting in the host (i.e., the reduction in outcrossing fitness through male function due to the selfing allele). We then sought the equilibrium level of host selfing under the different conditions. The results show that, over a wide range of conditions, parasites can select for host reproductive strategies in which both selfed and outcrossed progeny are produced (mixed mating). In addition, mixed mating, where it exits, tends to be biased toward selfing. PMID- 11430648 TI - The consequences of dioecy for seed dispersal: modeling the seed-shadow handicap. AB - Recent evidence has suggested that clades of dioecious angiosperms have fewer extant species on average than those of cosexual (hermaphroditic and monoecious) relatives. Reasons for the decrease in speciation rates and/or increase in extinction rates are only beginning to be investigated. One possibility is that dioecious species suffer a competitive disadvantage with cosexuals because only half of the individuals in a dioecious population are seed bearing. When only females produce seed, offspring will be more spatially clumped and will experience more local resource competition than when every individual produces seed. We examine two spatially explicit models to determine the effect of a reduction in seed dispersers on the invasibility and persistence of dioecious populations. Even though dioecious females were allowed to produce twice as many seeds as cosexuals, our results show that a reduction in the number of seed dispersers causes a decrease in the ability of dioecious progeny to find uninhabited sites, thus reducing persistence times. These results suggest that the maintenance of dioecy in the presence of hermaphroditic competitors requires a substantial increase in relative fitness and/or a large dispersal advantage of dioecious seeds. PMID- 11430649 TI - Evolution of thermal dependence of growth rate of Escherichia coli populations during 20,000 generations in a constant environment. AB - Twelve experimental populations of the bacterium Escherichia coli evolved for 20,000 generations in a defined medium at 37 degrees C. We measured their maximum growth rates across a broad range of temperatures and at several evolutionary time points to quantify the extent to which they became thermal specialists with diminished performance at other temperatures. We also sought to determine whether antagonistic pleiotropy (genetic trade-offs) or mutation accumulation (drift decay) was primarily responsible for any thermal specialization. Populations showed consistent improvement in growth rate at moderate temperatures (27-39 degrees C), but tended to have decreased growth rate at both low (20 degrees C) and high (41-42 degrees C) temperatures. Most loss occurred early in the experiment, when adaptation was most rapid. This dynamic is predicted by antagonistic pleiotropy but not by mutation accumulation. Several populations evolved high mutation rates due to defects in their DNA repair, but they did not subsequently undergo a greater decrease in growth rate at thermal extremes than populations that retained low mutation rates, contrary to the acceleration of decay predicted by mutation accumulation. Antagonistic pleiotropy therefore is more likely to be responsible for the evolution of thermal specialization observed in maximum growth rate. PMID- 11430650 TI - Variation of enzyme activities at a branched pathway involved in the utilization of gluconate in Escherichia coli. AB - Twenty-four strains of Escherichia coli from the ECOR collection were characterized for growth rate in gluconate minimal salts medium and for Vmax and Km of the three enzymes (gluconokinase, 6-phosphogluconate dehydrogenase, and 6 phosphogluconate dehydratase) that form a branch point for the utilization of gluconate. A total of 11 characters--growth rate, three Vmax values, four Km values, and three Vmax/Km values--were determined for these 24 ECOR strains. Most of the characters were normally distributed. Statistical tests showed that growth rate is significantly less variable than enzyme activities. Also, analyses of variance showed significant differences among strains and among the extant five genetic groups of E. coli for the characters measured. A Mantel test showed that, for some characters, closely related strains shared similar character values. Two hypotheses regarding the relationships between growth rate and enzyme activity and between various enzyme activities were tested. None of the expected correlations between growth rate and enzyme activity or between enzyme activities was detected. The results were discussed in terms of metabolic control analysis and neutral theory. PMID- 11430651 TI - Mutational meltdown in laboratory yeast populations. AB - In small or repeatedly bottlenecked populations, mutations are expected to accumulate by genetic drift, causing fitness declines. In mutational meltdown models, such fitness declines further reduce population size, thus accelerating additional mutation accumulation and leading to extinction. Because the rate of mutation accumulation is determined partly by the mutation rate, the risk and rate of meltdown are predicted to increase with increasing mutation rate. We established 12 replicate populations of Saccharomyces cerevisiae from each of two isogenic strains whose genomewide mutation rates differ by approximately two orders of magnitude. Each population was transferred daily by a fixed dilution that resulted in an effective population size near 250. Fitness declines that reduce growth rates were expected to reduce the numbers of cells transferred after dilution, thus reducing population size and leading to mutational meltdown. Through 175 daily transfers and approximately 2900 generations, two extinctions occurred, both in populations with elevated mutation rates. For one of these populations there is direct evidence that extinction resulted from mutational meltdown: Extinction immediately followed a major fitness decline, and it recurred consistently in replicate populations reestablished from a sample frozen after this fitness decline, but not in populations founded from a predecline sample. Wild-type populations showed no trend to decrease in size and, on average, they increased in fitness. PMID- 11430652 TI - Evolution of morphological novelty: a phylogenetic analysis of growth patterns in Streptocarpus (Gesneriaceae). AB - Streptocarpus shows great variation in vegetative architecture. In some species a normal shoot apical meristem never forms and the entire vegetative plant body may consist of a single giant cotyledon, which may measure up to 0.75 m (the unifoliate type) or with further leaves arising from this structure (the rosulate type). A molecular phylogeny of 87 taxa (77 Streptocarpus species, seven related species, and three outgroup species) using the internal transcribed spacers and 5.8S region of nuclear ribosomal DNA suggests that Streptocarpus can be divided into two major clades. One of these broadly corresponds to the caulescent group (with conventional shoot architecture) classified as subgenus Streptocarpella, whereas the other is mainly composed of acaulescent species with unusual architecture (subgenus Streptocarpus). Some caulescent species (such as S. papangae) are anomalously placed with the acaulescent clade. Available cytological data are, however, completely congruent with the two major clades: the caulescent clade is x = 15 and the acaulescent clade (including the caulescent S. papangae) is x = 16 (or polyploid multiples of 16). The genera Linnaeopsis, Saintpaulia, and Schizoboea are nested within Streptocarpus. The sequenced region has evolved, on average, 2.44 times faster in the caulescent clade than in the acaulescent clade and this is associated with the more rapid life cycle of the caulescents. Morphological variation in plant architecture within the acaulescent clade is homoplastic and does not appear to have arisen by unique abrupt changes. Instead, rosulate and unifoliate growth forms have evolved several times, reversals have occurred, and intermediate architectures are found. An underlying developmental plasticity seems to be a characteristic of the acaulescent clade and is reflected in a great lability of form. PMID- 11430653 TI - Phenotypic plasticity early in life constrains developmental responses later. AB - Plastic stem-elongation responses to the ratio of red:far-red (R:FR) wavelengths enable plants to match their phenotype to local competitive conditions. However, elongation responses early in the life history may occur at the cost of reduced plasticity later in the life history, because elongation influences both allocation patterns and structural integrity. A common-garden experiment was performed to test whether seedling responses to R:FR affect biomass allocation. biomass accumulation, and subsequent plasticity to the cue. Seedlings of Abutilon theophrasti were stimulated to elongate by low R:FR treatments, and subsequent growth and plasticity was compared with nonelongated individuals. Elongated seedlings were less responsive than nonelongated ones to a second bout of low R:FR. Thus, seedling plasticity to R:FR reduces subsequent responsiveness to this cue. This negative association across life-history stages suggests an important constraint on the evolution of plastic stem responses, because selection in A. theophrasti has previously been shown to favor increases in early elongation in combination with increased later elongation. The reduced responsiveness of elongated seedlings to R:FR appeared to result from a structural feedback mechanism, indicating that the opportunity cost of early responses may be lower in environments providing structural support. PMID- 11430654 TI - Deleterious mutations and genetic variation for flower size in Mimulus guttatus. AB - Mimulus guttatus is a wildflower that exhibits substantial genetic variation in flower size. Here, we test the hypothesis that this variation is caused by deleterious mutations maintained through mutation-selection balance. The deleterious-mutation model predicts that rare, partially recessive alleles will be the primary source of variation. We test this prediction by measuring the change in the mean flower size (deltaM) and the directional dominance of flower size (deltaB) within a selection experiment. If variation is due to rare (partially) recessive alleles, deltaB/deltaM is expected to be positive and exceed one. However, we obtain negative values for deltaB/deltaM from three independent selection lines. This result is statistically inconsistent with the deleterious-mutation model. PMID- 11430655 TI - Patterns of allozyme variation in diploid and tetraploid Centaurea jacea at different spatial scales. AB - The extent and spatial patterns of genetic variation at allozyme markers were investigated within and between diploid and autotetraploid knapweeds (Centaurea jacea L. sensu lato, Asteraceae) at contrasted geographic scales: (1) among populations sampled from a diploid-tetraploid contact zone in the northeastern part of the Belgian Ardennes, and (2) within mixed populations from that zone where diploids and tetraploids coexist. Our data were also compared with a published dataset by Sommer (1990) describing allozyme variation in separate diploid and tetraploid knapweeds populations collected throughout Europe. Genetic diversity was higher in tetraploids. In the Belgian Ardennes and within the mixed populations, both cytotypes had similar levels of spatial genetic structure, they were genetically differentiated, and their distributions of allele frequencies were not spatially correlated. In contrast, at the European scale, diploids and tetraploids did not show differentiated gene pools and presented a strong correlation between their patterns of spatial genetic variation. Numerical simulations showed that the striking difference in patterns observed at small and large geographic scales could be accounted for by a combination of (1) isolation by distance within cytotypes; and (2) partial reproductive barriers between cytotypes and/or recurrent formation of tetraploids. We suggest that this may explain the difficulty of the taxonomic treatment of knapweeds and of polyploid complexes in general. PMID- 11430656 TI - Population structure and speciation in tropical seas: global phylogeography of the sea urchin Diadema. AB - The causes of speciation in the sea are rarely obvious, because geographical barriers are not conspicuous and dispersal abilities or marine organisms, particularly those of species with planktonic larvae, are hard to determine. The phylogenetic relations of species in cosmopolitan genera can provide information on the likely mode of their formation. We reconstructed the phylogeny of the pantropical and subtropical sea urchin genus Diadema, using sequences of mitochondrial DNA from 482 individuals collected around the world, to determine the efficacy of barriers to gene flow and to ascertain the history of possible dispersal and vicariance events that led to speciation. We also compared 22 isozyme loci between all described species except D. palmeri. The mitochondrial DNA data show that the two deepest lineages are found in the Indian and West Pacific Oceans. (Indo-Pacific) Diadema setosum diverged first from all other extant Diadema, probably during the initiation of wide fluctuations in global sea levels in the Miocene. The D. setosum clade then split 3-5 million years ago into two clades, one found around the Arabian Peninsula and the other in the Indo-West Pacific. On the lineage leading to the other species of Diadema, the deepest branch is composed of D. palmeri, apparently separated when the climate of New Zealand became colder and other tropical echinoids at these islands went extinct. The next lineage to separate is composed of a currently unrecognized species of Diadema that is found at Japan and the Marshall Islands. Diadema mexicanum in the eastern Pacific separated next, whereas D. paucispinum, D. savignyi, and D. antillarum from the western and central Atlantic, and (as a separate clade) D. antillarum from the eastern Atlantic form a shallow polytomy. Apparently, Indo Pacific populations of Diadema maintained genetic contact with Atlantic ones around the southern tip of Africa for some time after the Isthmus of Panama was complete. Diadema paucispinum contains two lineages: D. paucispinum sensu stricto is not limited to Hawaii as previously thought, but extends to Easter Island, Pitcairn, and Okinawa; A second mitochondrial clade of D. paucispinum extends from East Africa and Arabia to the Philippines and New Guinea. A more recent separation between West Indian Ocean and West Pacific populations was detected in D. setosum. Presumably, these genetic discontinuities are the result of water flow restrictions in the straits between northern Australia and Southeast Asia during Pleistocene episodes of low sea level. Diadema savignyi is characterized by high rates of gene flow from Kiribati in the central Pacific all the way to the East African Coast. In the Atlantic, there is a biogeographic barrier between the Caribbean and Brazil, possibly caused by fresh water outflow from the Amazon and the Orinoco Rivers. Diadema antillarum populations of the central Atlantic islands of Ascension and St. Helena are genetically isolated and phylogenetically derived from Brazil. Except for its genetic separation by the mid-Atlantic barrier, Diadema seems to have maintained connections through potential barriers to dispersal (including the Isthmus of Panama) more recently than did Eucidaris or Echinometra, two other genera of sea urchins in which phylogeography has been studied. Nevertheless, the mtDNA phylogeography of Diadema includes all stages expected from models of allopatric differentiation. There are anciently separated clades that now overlap in their geographic distribution, clades isolated in the periphery of the genus range that have remained in the periphery, clades that may have been isolated in the periphery but have since spread towards the center, closely related clades on either side of an existing barrier, and closely related monophyletic entities on either side of an historical barrier that have crossed the former barrier line, but have not attained genetic equilibrium. Except for D. paucispinum and D. savignyi, in which known hybridization may have lodged mtDNA from one species into the genome of the other, closely related clades are always allopatric, and only distantly related ones overlap geographically. Thus, the phylogenetic history and distribution of extant species of Diadema is by and large consistent with allopatric speciation. PMID- 11430657 TI - Population genetics and colony structure of the Argentine ant (Linepithema humile) in its native and introduced ranges. AB - Introduced species often possess low levels of genetic diversity relative to source populations as a consequence of the small population sizes associated with founder events. Additionally, native and introduced populations of the same species can possess divergent genetic structuring at both large and small geographic scales. Thus, genetic systems that have evolved in the context of high diversity may function quite differently in genetically homogeneous introduced populations. Here we conduct a genetic analysis of native and introduced populations of the Argentine ant (Linepithema humile) in which we show that the population-level changes that have occurred during introduction have produced marked changes in the social structure of this species. Native populations of the Argentine ant are characterized by a pattern of genetic isolation by distance, whereas this pattern is absent in introduced populations. These differences appear to arise both from the effects of recent range expansion in the introduced range as well as from differences in gene flow within each range. Relatedness within nests and colonies is lower in the introduced range than in the native range as a consequence of the widespread genetic similarity that typifies introduced populations. In contrast, nestmates and colony-mates in the native range are more closely related, and local genetic differentiation is evident. Our results shed light on the problem posed for kin selection theory by the low levels of relatedness that are characteristic of many unicolonial species and suggest that the loss of genetic variation may be a common mechanism for the transition to a unicolonial colony structure. PMID- 11430658 TI - Signal polymorphism in the web-decorating spider Argiope argentata is correlated with reduced survivorship and the presence of stingless bees, its primary prey. AB - Many spiders, and in particular those in the genus Argiope, spin highly visible web decorations whose function and significance are the subject of spirited debate. In this work, we present data to address two of the competing hypotheses that fuel this controversy. In particular, we examine the relationship between the presence of web decorations and spider survivorship (predator-protection hypothesis) and the relationship between the presence of prey and spider decorating behavior (the prey-attraction hypothesis). Our laboratory studies reveal that the decorating behavior of the spider A. argentata has a genetic component but that the expression of decorating behavior tends to be elicited only when a spider is well fed. Furthermore, our field studies show that in the presence of abundant stingless bees, spider decorating behavior is induced. Nevertheless, our field surveys also suggest that spiders that decorate their webs show reduced survivorship. We propose that the high correlation between web decorating in the presence of stingless bees supports the hypothesis that A. argentata engage in decorating behavior when attracting or targeting specific prey types. However, we also propose that web decorations attract the predators of A. argentata because high-frequency decorators suffer lower survivorship than spiders that decorate moderately or rarely. These findings suggest that spider web decorating behavior is affected by conflicting selection pressures: the positive effect of prey attraction versus the negative effect of predator attraction. Due to the heritable component of decorating behavior, web decorating among A. argentata is likely to be particularly sensitive to the spider's local ecology as well as local patterns of gene flow. PMID- 11430659 TI - Females receive a life-span benefit from male ejaculates in a field cricket. AB - Mating has been found to be costly for females of some species because of toxic products that males transfer to females in their seminal fluid. Such mating costs seem paradoxical, particularly for species in which females mate more frequently than is necessary to fertilize their eggs. Indeed, some studies suggest that females may benefit from mating more frequently. The effect of male ejaculates on female life span and lifetime fecundity was experimentally tested in the variable field cricket, Gryllus lineaticeps. In field crickets, females will mate repeatedly with a given male and mate with multiple males. Females that were experimentally mated either repeatedly or multiply lived more than 32% longer than singly mated females. In addition, multiply mated females produced 98% more eggs than singly mated females. Because females received only sperm and seminal fluid from males in the experimental matings, these life-span and fecundity benefits may result from beneficial seminal fluid products that males transfer to females during mating. Mating benefits rather than mating costs may be common in many animals, particularly in species where female mate choice has a larger effect on male reproductive success than does the outcome of sperm competition. PMID- 11430660 TI - The development, validity and application of a new instrument to assess the quality of discharge planning activities from the community perspective. AB - OBJECTIVE: To describe the development, validity and application of a new instrument (PREPARED) for obtaining feedback from community consumers of discharge planning activities. DESIGN: Iterative qualitative and quantitative investigations. SETTING: The community catchment area of a metropolitan Australian tertiary public hospital. STUDY PARTICIPANTS: Patients aged over 65 years, with a range of conditions, recently discharged from hospital, their carers, and hospital nursing staff. ACTIONS: PREPARED was constructed from interviews with patients, carers and hospital staff. It was trialed and modified to ensure sensitive measurement of key attributes of discharge planning process and outcome. This paper explores the patient and carer versions of PREPARED. Data items were reduced to domains of key questions by factor analysis. Instrument performance was assessed by correlation of process and outcome measures, by comparing PREPARED responses with subsequent unstructured interview data, and by testing whether PREPARED responses were independent of health-related quality of life at the time of survey. RESULTS: Four key process domains were identified: information exchange (community services and equipment), medication management, preparation for coping after discharge and control of discharge circumstances. Outcome was measured as overall satisfaction with discharge, whether equipment and community service needs had been met, use of health services and health related costs post-discharge. The instrument performed well when compared with interview data, the process and outcome domains were largely independent of each other, as were responses to PREPARED and SF-36. CONCLUSIONS: PREPARED offers a comprehensive way of closing the quality improvement loop, by providing information from the community perspective on the quality of planning for discharge from the acute hospital setting. PMID- 11430661 TI - Social class inequalities in the use of and access to health services in Catalonia, Spain: what is the influence of supplemental private health insurance? AB - OBJECTIVE: To analyse social class inequalities in the access to and utilization of health services in Catalonia (Spain), and the influence of having private health insurance supplementing the National Health System (NHS) coverage. DESIGN: 1994 Catalan Health Interview Survey, a cross-sectional survey conducted in 1994. SETTING: Catalonia (Spain). STUDY PARTICIPANTS: The participants were a representative sample of people aged over 14 years from the non-institutionalized population of Catalonia (n = 12,245). MAIN OUTCOME MEASURES: Health services utilization, perceived health, having only NHS or NHS plus a private health insurance, and social class. RESULTS: Although one-quarter of the population of Catalonia had a supplemental private health insurance, percentages were very different according to social class, ranging from almost 50% for classes I and II to 16% for classes IV and V in both sexes. No inequalities by social class were observed for the utilization of non-preventive health care services (consultation with a health professional in the last 2 weeks and hospitalization in the last year) among persons with poor self-perceived health status, i.e. those in most need. However, social inequalities still remain in the use of health services provided only partially by the NHS, and when characteristics of last consultation are taken into account. Subjects who paid for a private service waited an average of 18.8 minutes less than those attending the NHS. Within the NHS, social classes IV and V waited longer (35.5 minutes) than social classes I and II (28.4 minutes). CONCLUSION: The NHS in Catalonia, Spain, has reduced inequalities in the use of health services. Social inequalities remain in the use of those health services provided only partially by the NHS. PMID- 11430662 TI - Applicability of diagnostic recommendations on dementia in family practice. AB - OBJECTIVE: To evaluate the applicability of the Dutch dementia guideline's recommendations, including the diagnostic criteria used by family practitioners, and to explore characteristics in both patients and family practitioners which are associated with the use of these recommendations. DESIGN: An observational study was set up with a sample of 64 family practitioners who were instructed to use the Dutch national dementia guideline on incident-suspected dementia patients. The applicability was expressed as the percentage of recommendations applied. The use of diagnostic criteria was checked by comparing the family practitioners diagnoses with the diagnoses received by integrating the registered symptoms according to the DSM-III-R criteria. Associations between the number of recommendations applied, and demographic and clinical features were explored. MAIN OUTCOME MEASURES: (i) guideline applicability (ii) integrated use of DSM-III R criteria. RESULTS: 107 patients were included. The average application rate of the guideline's 31 diagnostic key recommendations was 86% or 24.8 (SD 3.6). The family practitioners diagnoses were consistent with the expected DSM-III-R diagnoses in 26% of the cases (kappa = 0.1). A greater number of patients in a practice was positively associated with the use of recommendations. A need for referral by the family practitioners, and patients' denial of dementia were negatively associated with the use of recommendations. The presence of dementia and a patient's age were negatively associated with the use of the DSM-III-R criteria. CONCLUSIONS: The applicability of the diagnostic recommendations of the national Dutch dementia guideline in a representative sample of family practitioners was promising. Nevertheless, the diagnostic criteria of the DSM-III R, which were part of the dementia guideline, provided little or no guidance to the family practitioners in their diagnostic decision-making. Clinical and demographic variables explained some of the variation in the use of recommendations. PMID- 11430663 TI - Completing the audit cycle: the outcomes of audits in mental health services. AB - AIMS: To assess how far those UK National Health Service mental health settings that tested, and prior to publication, used the Newcastle Clinical Audit Toolkit for Mental Health (NCAT) completed the audit cycle. DATA SOURCES: Twelve clinical audit project reports, each focused on one of the five modules in the NCAT, from four rounds of activity over a 2-year period; clinical and managerial staff in the settings where audit projects had taken place. DATA EXTRACTION: Interviews with audit project team members about the recommendations of the 12 audit project reports and about contextual issues; all projects had reported at least 2 years previously. RESULTS OF DATA SYNTHESIS: In analysing the audit project outcomes, five categories of inaction were discernible and five further categories were needed to describe varying states of progress. It was necessary to discriminate between actions attributed to the NCAT audit projects and actions attributed mainly to other initiatives. In total, 26.4% of audit recommendations were still under discussion or in progress. A relatively low proportion of recommendations from audit report findings (34.7%) had been implemented, and these were divided almost equally between recommendations attributed to the NCAT projects (38) and those attributed to other initiatives in the organization (37). CONCLUSION: Investigation of the medium-term outcomes of clinical audit projects has provided an insight into what might usefully be termed the process of completing the audit cycle. The time-scales required to reach the point at which action is deemed to have been implemented or not may be as long as 3 years. Conceptualizing the action stage of the cycle as a single discrete event fails to do justice to the complexity of the process, and attributing the implementation of change in clinical settings to single causes such as individual audit projects is problematic. PMID- 11430664 TI - Regional guideline for diagnosis and treatment of squamous cell carcinoma of the lip: what is the level of compliance? AB - OBJECTIVE: To investigate to what extent physicians comply with a regional guideline for the diagnosis, staging, treatment and follow-up of patients with squamous cell carcinoma of the lower lip. DESIGN: Retrospective analysis of data from the medical records of 248 patients diagnosed with squamous cell carcinoma of the lower lip during the period 1989-1997. SETTING: Comprehensive Cancer Centre of the Northern region of the Netherlands. STUDY PARTICIPANTS: The data were collected by this Centre for the regional population-based cancer registry. RESULTS: Overall compliance with the separate guidelines varied between 4 and 80%. For diagnosis and staging, the guideline was followed for 4-70% of patients. The type of treatment in relation to age conformed to the guideline in 34% of cases. Of the 208 surgically-treated patients, treatment was performed in accordance with the guidelines in 92 (44%) patients, compared with seven out of 18 (39%) patients who received radiotherapy. Follow-up in accordance with the guideline was 11% in the first year, 9% in the second year and 21% in the third year. CONCLUSION: Only a minority of patients with squamous cell carcinoma of the lower lip, a rare rumour, were managed according to the available regional guideline. Regular review of both the guideline and its implementation is necessary in order to optimize its use. PMID- 11430666 TI - Medication error prevention: potassium chloride. PMID- 11430665 TI - Statistical presentation. PMID- 11430667 TI - Quality programmes, accreditation and certification in Switzerland. AB - PURPOSE: To report on current quality initiatives in health care in the context of accreditation and certification in Switzerland. SOURCE OF INFORMATION: A review from the Swiss Accreditation Service, the Swiss Medical Association and the University of Zurich. SETTING: In Switzerland, health care providers and health insurers are obliged by law to sign contracts on quality of care. The law texts and related ordinances do not state explicitly the content and format of quality contracts between providers and costs payers. Finding adequate practical solutions is the responsibility of the partners involved. Six different possibilities of quality initiatives are studied and discussed in this report. These include initiatives to create independent certification or inspection bodies for health care organizations accredited by the national accreditation body. FINDINGS: So far, there is only one established standardized system with clear requirements. This accreditation and certification model is an adaptation of the International Standards Organization but so far it has not been used often in the public domain. Each of the other five approaches are more frequently applied but none of them lead to full external peer review certification including medical outcome yet. CONCLUSIONS: A lot of harmonization work still has to be done and clear branch specific requirements need to be agreed upon. Practicability and validity of external peer review schemes may also need further evaluation and improvement to reach the goals of health care systems in the future. PMID- 11430668 TI - Improving the quality of health care in Latin America. PMID- 11430669 TI - Evaluation of 60 continuous quality improvement projects in French hospitals. AB - OBJECTIVE: To evaluate the feasibility of implementing continuous quality improvement (CQI) projects in French health care organizations. DESIGN: The French Ministry of Health issued two calls for CQI projects (in 1995 and 1996). ANAES was commissioned to monitor and evaluate the projects, and to provide advice. SETTING: ANAES in collaboration with French public hospitals. STUDY PARTICIPANTS: A jury selected 64 projects from 483 submissions. The first series of projects related to safety issues (e.g. blood transfusions), the second related chiefly to patient management. INTERVENTIONS: ANAES instructed project leaders in process analysis (modified four-step FOCUS-PDCA model), convened regular meetings between leaders and performed on-site visits. MAIN OUTCOME MEASUREMENTS: Objective outcomes: goal achievement, extension of projects to other topics and departments, allocation of resources. Subjective outcomes: changes in attitudes. Statistics were obtained from two questionnaires completed by project leaders. RESULTS: Four projects were discontinued; 82% (49 out of 60) met more than half their objectives. The CQI method was adopted by other departments in 65% and 50% (1st and 2nd series respectively) of cases. Hospital management often chose to provide continued support (81%/88%), offer training (59%/80%), create a CQI unit (62%/73%), and allocate a budget (61%/65%). A positive impact on staff attitudes was noted in over 75% of projects. CONCLUSION: ANAES' co-ordinated initiative to acquaint a hard core of French public hospitals with CQI proved successful. Identification of the factors for success and of potential hurdles helped pave the way for the national hospital accreditation procedure currently underway. PMID- 11430670 TI - Development of a French inpatient satisfaction questionnaire. AB - OBJECTIVE: To develop a brief French-language, generic, self-administered questionnaire to measure inpatient satisfaction. DESIGN: Issues relevant to patients were identified using three open-ended questions designed in accordance with the disconfirmation paradigm. The content of patients' responses was analysed and then supplemented by items taken from published instruments in order to generate a pool of 93 items. Twenty-nine items were selected following a strict procedure. Content validity was judged by comparing the questionnaire to existing instruments. Construct validity was supported by testing specific hypotheses derived from the literature and by performing principal component analysis. Reliability was estimated by calculating Cronbach's alpha. SETTING: A 2200-bed French teaching hospital. SUBJECTS: A mail survey was carried out on a random sample of 1000 inpatients within 2-4 weeks of discharge. Eligible subjects were medical, surgical and obstetrics inpatients who had stayed in the hospital for more than 24 hours. RESULTS: The participation rate (71%) and the completion rate (95%) were indicators of acceptability. There were modest differences between the questionnaire and published instruments (financial aspects, amenities). Construct representation by principal component analysis consisted of six scales which accounted for 58% of the variance in total satisfaction scores. The reliability estimates of internal consistency ranged from 0.67 to 0.86. CONCLUSION: We propose that the self-administered multidimensional inpatient satisfaction questionnaire provided encouraging preliminary psychometric information. This instrument is intended to involve patient feedback in a continuous quality health care improvement strategy. PMID- 11430671 TI - Radiation monitoring in Russia at the turn of the millennium. PMID- 11430672 TI - Natural radionuclides as tracers of the dispersal of dredge spoil dumped at sea. AB - Monitoring large (underwater) surfaces, with rapidly varying composition, requires a sampling density which exceeds the capabilities of standard techniques. These techniques involve sample collection and a number of treatments and measurements in the laboratory; both steps are laborious, tedious and costly. This paper presents an in situ method in which a detector system is trailed over the surface and measures continuously the gamma rays emitted by the natural radionuclides in the sediment. Since each sediment component has its own characteristic set of activity-concentration values (radiometric fingerprint), the composition of the sediment can be deduced quantitatively. This paper shows the application of this technique for monitoring the dispersal of dredge spoil from Rotterdam harbour, dumped in the North Sea. In addition to a qualitative picture of dredge spoil dispersal, a mass balance equation has been used to quantitatively assess the dredge spoil transport with time. PMID- 11430673 TI - Sorption behaviour of Am on marl groundwater colloids. AB - The sorption of americium on marl colloids is studied in a comprehensive way. In Na-HCO3 groundwater conditions, americium in the concentration range 3 x 10(-11) 3 x 10(10) M sorbs onto groundwater colloids. In these tests, the colloid concentration is rather high, i.e. Ccoll 1.7+/-1.3 mg l(-1) for sizes 505000 nm or Ncoll (1.7+/-0.1) x 10(11) l(-1) for sizes larger than 100 nm. These colloids are mainly chlorite and smectite/illite with an average size smaller than 500 nm. For the groundwater (pH 8.6, [Na+]-[HCO3]-2 x 10(-2) M), Kd values in the range 1 3 x 10(6)mlg(-1) are measured. These values are compared with experimental and modelled values reported earlier in the literature. PMID- 11430674 TI - Radiological impacts of natural radioactivity in Abu-Tartor phosphate deposits, Egypt. AB - Phosphate and environmental samples were collected from Abu Tartor phosphate mine and the surrounding region. The activity concentration of 226Ra (238U) series, 232Th series and 40K were measured using a gamma-ray spectrometer. The activities of uranium isotopes (238U, 235U and 234U) and 210Pb were measured using an alpha spectrometer and a low-background proportional gas counting system, respectively, after radiochemical separation. The results are discussed and compared with the levels in phosphate rocks from different countries. It seems that the Abu Tartor phosphate deposit has the lowest radioactivity level of exploited phosphate of sedimentary origin. 226Ra/238U, 210Pb/226Ra, 234U/238U and 226Ra/228Ra activity ratios were calculated and are discussed. The radioactivity levels in the surrounding region and the calculated exposure dose (nGy/h) will be considered as a pre-operational baseline to estimate the possible radiological impacts due to mining, processing and future phosphate industrial activities. To minimize these impacts, the processing wastes should be recycled to the greatest possible extent. PMID- 11430675 TI - Wet and dry deposition of 129I in Seville (Spain) measured by accelerator mass spectrometry. AB - Iodine-129 (T1/2 = 1.57 x 10(7) yr) concentrations have been determined by accelerator mass spectrometry in rainwater samples taken at Seville (southwestern Spain) in 1996 and 1997. This technique allows a reduction in the detection limits for this radionuclide in comparison to radiometric counting and other mass spectrometric methods such as ICP-MS. Typical 129I concentrations range from 4.7 x 10(7) 129I atoms/l (19.2%) to 4.97 x 10(9) 129I atoms/l (5.9%), while 129I depositions are normally in the order of 10(8)-10(10) atoms/m2d. These values agree well with other results obtained for recent rainwater samples collected in Europe. Apart from these, the relationship between 129I deposition and some atmospheric factors has been analyzed, showing the importance of the precipitation rate and the concentration of suspended matter in it. PMID- 11430676 TI - Seasonal variations in radionuclide transfer in a Mediterranean grazing-land ecosystem. AB - We made quarterly determinations of the transfer coefficients and effective transfer coefficients for the radionuclides 137Cs, 90Sr, 40K, 226Ra, 228Ra and 228Th over a full annual cycle, in a Mediterranean grazing-land ecosystem. The input and output fluxes of the radionuclides between the different compartments of this ecosystem were quantified for the following processes: root uptake; variation in root and aerial biomass; pasture production; translocation; leaf fall; efflux due to grazing action; resuspension and subsequent aerial deposition of radionuclides. We observed there to be a marked seasonal variation for this type of ecosystem in both the transfer coefficients and the radionuclide fluxes, which impedes the soil-plant transfer being characterized on the basis of values that are constant with time. PMID- 11430677 TI - Effect of external potassium (K) supply on the uptake of 137Cs by spring wheat (Triticum aestivum cv. Tonic): a large-scale hydroponic study. AB - A large-scale hydroponic experiment was carried out in a non-controlled greenhouse. Spring wheat plants were grown to maturity at four levels of external K concentration (2, 4, 20 and 40 mgl(-1)) and one concentration of radiocaesium (8 Bqml(-1)). Concentrations of K and radiocaesium in the growth solution were closely monitored, and replenishments were made upon depletion. K effectively competed with radiocaesium in terms of root uptake. Activity concentrations of radiocaesium in plants differed significantly between the four K treatments; the activity concentration at the lowest external K concentration being 100 times higher than that at the highest K level. The relationship between radiocaesium uptake and external K level could be described by a negative power function; this showed that when the K level reached around 12 mgl(-1), further increases in the external K level resulted only in slight changes in its inhibitory effect. As a result of this inhibitory effect of potassium supply, concentrations of radiocaesium in plant tissues, grains in particular, were greatly reduced at high external K concentration. Mechanisms involved in Cs-K interaction in root uptake are also discussed. PMID- 11430678 TI - A comparison of three models of 137Cs transfer in forest ecosystems. AB - The predictions of three models of 137Cs transfer in forest ecosystems (FOA, LOGNAT and FORESTLAND) were compared. The scenario for the model-model comparison consisted of an acute dry deposition of 137Cs over a coniferous forest. The model predictions were subsequently compared (model-data comparison) with values derived from experimental data measured in forests of the Bryansk region in Russia that were contaminated by the Chernobyl accident and that have similar characteristics to the forests described in the scenario. The predictions of radiocaesium levels in the litter-soil layer, berries, needles, wood, whole tree and moose made with the models were in relatively good agreement with each other (within a factor of 1.4-2.9). The best agreement was observed for berries and moose and the worst for wood. There was also good agreement between the model predictions for the same variables and the experimental data (within a factor of 1.2 3.2). In this case, the best agreement was observed for the litter-soil layer and the worst for wood and the whole tree. Overall, at least for the studied scenario and for the first 10 years after deposition, any of the models can be used if the final aim is to estimate average concentrations in different forest components. The agreement between the model predictions worsens with time and there were differences in the form of the time dependencies predicted by the models, especially for wood. This may lead to larger differences between the model predictions and the experimental data for times beyond the period for which data were available for comparison (10 years after the deposition). PMID- 11430679 TI - Genetically modified animals: the Royal Society considers costs and benefits. PMID- 11430680 TI - Use of breed-specific ranges for the vertebral heart scale as an aid to the radiographic diagnosis of cardiac disease in dogs. AB - The vertebral heart scale was measured on right lateral recumbent thoracic radiographs of 320 dogs of six popular breeds, including for each breed at least 20 dogs with no clinical signs of cardiovascular or respiratory disease and at least 19 dogs with cardiac or respiratory disease. There were significant differences between the mean values of the scale for the different breeds; the normal boxer dogs had a significantly higher mean value than the normal dogs of all the other breeds, and the labrador retrievers had a significantly higher mean value than all the other breeds except the boxer and the cavalier King Charles spaniel. For all the breeds except the boxer, there was a trend for dogs with cardiac disease (but not respiratory disease) to have higher mean values on the scale than normal dogs of the same breed; however, at the optimal value of the scale for distinguishing between dogs of each breed with and without cardiac disease, the sensitivity and specificity were relatively low, in the range 58 to 83 per cent. The scale was most accurate for the diagnosis of cardiac disease in the Yorkshire terrier and the cavalier King Charles spaniel, breeds affected by predominantly dilative forms of cardiac disease. In contrast, it was very inaccurate in the boxer, a breed that has a higher incidence of cardiac diseases associated with concentric hypertrophy. PMID- 11430681 TI - Relationships between infection with caprine arthritis encephalitis virus, intramammary bacterial infection and somatic cell counts in dairy goats. AB - Somatic cell counts, the bacteriological condition of the milk and antibodies against caprine arthritis encephalitis virus (CAEV) were measured monthly throughout lactation in 121 lactating goats of the Murcia-Granada breed in four commercial dairy goat herds. The prevalence of bacterial intramammary infection was 5.6 per cent and the prevalence of CAEV infection was 20.6 per cent. An analysis of variance revealed a significant effect of herd, intramammary infection and the interaction between intramammary infection and CAEV on the somatic cell count. In udder halves free of intramammary infection, the somatic cell counts were significantly lower in seronegative goats than in seropositive goats (P<0.05), but the difference was not significant in udder halves persistently infected by bacteria. There was a significant increase in somatic cell counts due to bacterial intramammary infection (P<0.01) in the seronegative goats, but this effect was not present in the seropositive animals. PMID- 11430682 TI - Evidence of infectious diseases in harbour porpoises (Phocoena phocoena) hunted in the waters of Greenland and by-caught in the German North Sea and Baltic Sea. AB - The pathological, microbiological and serological findings in harbour porpoises hunted in Greenlandic waters were compared with the findings in animals accidentally caught in fishing gear in the German North Sea and Baltic Sea. The body condition of the Greenlandic animals was good, whereas nine of 23 German harbour porpoises were moderately to markedly emaciated. Both groups were infested with parasites. In the Greenlandic animals parasitism of the aural peribullar cavity with Stenurus minor, of the liver and pancreas with Orthosplanchnus mironovi, of the lungs with Halocercus species and of the subcutaneous and mammary tissue with Crassicauda species was generally associated with a mild inflammatory response. No diseases associated with bacteria were identified in any of the Greenlandic harbour porpoises. In the porpoises from the German North Sea and Baltic Sea, parasites were present in the aural peribullar cavity (S minor), liver (Campula oblonga), first and second gastric compartment (Anisakis simplex) and in the lungs (Pseudalius inflexus and Torynurus convolutus). Moderate to marked pulmonary parasitism and suppurative pneumonia, not observed in the Greenlandic porpoises, were present in 11 and 10, respectively, of the 23 German porpoises. The suppurative pneumonia was attributed to bacterial infection with beta-haemolytic streptococci and Escherichia coil var haemolytica. Four Greenlandic and 10 German porpoises had positive porpoise morbillivirus-specific antibody titres suggesting that the virus was circulating in both populations. PMID- 11430683 TI - Oral lesions in sheep and cattle in Dumfries and Galloway. PMID- 11430684 TI - Activity of oxyclozanide, nitroxynil, clorsulon and albendazole against adult triclabendazole-resistant Fasciola hepatica. PMID- 11430685 TI - Cowdriosis (heartwater) in a Ugandan goat: a case report. PMID- 11430686 TI - FMD controls: RCVS advice on signing Form A. PMID- 11430687 TI - Veterinary school league tables. PMID- 11430688 TI - Canine granulocytic ehrlichiosis in the UK. PMID- 11430689 TI - PDSA review. PMID- 11430690 TI - PDSA review. PMID- 11430691 TI - Vascular alterations in the rabbit patellar tendon after surgical incision. AB - Open incision of the patellar tendon (PT) is thought to promote acute vascular responses which ultimately result in an enhanced degree of tendon repair. Such a clinical procedure is commonly applied to patients with refractory tendinitis. The objective of this study was to quantify the vascular adaptations (both anatomical and physiological) to longitudinal incision of the PT, and the resultant effects on tendon organisation. Fifty-four New Zealand White rabbits were separated into 3 experimental groups and 2 control groups. Experimental groups underwent surgical incision of the right PT, and were assessed 3 d, 10 d and 42 d following injury; normal unoperated controls were evaluated at time zero, and sham-operated controls were evaluated at 3 d to control for the effects of incising the overlying skin. Quantitative measures of PT blood supply (blood flow, microvascular volume) and geometric properties of PT substance were obtained for each PT. Histomorphology was assessed to evaluate vascular remodelling and matrix organisation in the healing PT. Longitudinal open incision surgery of the PT led to rapid increases in both blood flow and vascular volume. The incision of overlying tissues alone (sham-operated) contributed to this measurable increase, and accounted for 36% and 42% of the elevated blood flow and vascular volume respectively at the 3 d interval. In the incised PT, blood flow significantly increased by 3 d compared with both time zero and sham-operated controls, and remained significantly elevated at the 10 d interval. Similarly, vascular volume of the incised PT increased at 3 d compared both with time zero and sham-operated controls. At the 10 d interval, the increase in vascular volume was greatest in the central PT substance. By 42 d both blood flow and vascular volume of the incised tendon had diminished, with only blood flow remaining significantly different from controls. In the contralateral limb, a significant neurogenically mediated vasodilation was measured in the contralateral PTs at both early time intervals, but was not seen by the later 42 d interval. With respect to PT geometric properties in the experimental animals, a larger PT results as the tendon matrix and blood vessels remodel. PT cross-sectional area increased rapidly by 3 d to 1.3 times control values, and remained significantly elevated at 42 d postinjury. Morphological assessments demonstrated the disruption of matrix organisation by vascular and soft tissue components associated with the longitudinal incisions. Substantial changes in matrix organisation persisted at 42 d after surgery. These findings suggest that open longitudinal incision of the PT increases the vascular supply to deep tendon early after injury. These changes probably arise through both vasomotor and angiogenic activity in the tissue. Since PT blood flow and vascular volume return towards control levels after 6 wk but structural features remain disorganised, we propose that vascular remodelling is more rapid and complete than matrix remodelling after surgical incision of the PT. PMID- 11430692 TI - The cells of the rabbit meniscus: their arrangement, interrelationship, morphological variations and cytoarchitecture. AB - Four major morphologically distinct classes of cells were identified within the adult rabbit meniscus using antibodies to cytoskeletal proteins. Two classes of cell were present in the fibrocartilage region of the meniscus. These meniscal cells exhibited long cellular processes that extended from the cell body. A third cell type found in the inner hyaline-like region of the meniscus had a rounded form and lacked projections. A fourth cell type with a fusiform shape and no cytoplasmic projections was found along the superficial regions of the meniscus. Using a monoclonal antibody to connexin 43, numerous gap junctions were observed in the fibrocartilage region, whereas none were seen in cells either from the hyaline-like or the superficial zones of the meniscus. The majority of the cells within the meniscus exhibited other specific features such as primary cilia and 2 centrosomes. The placement of the meniscal cell subtypes as well as their morphology and architecture support the supposition that their specific characteristics underlie the ability of the meniscus to respond to different types of environmental mechanical loads. PMID- 11430693 TI - Investigation of feline brain anatomy for the detection of cortical spreading depression with magnetic resonance imaging. AB - Cortical spreading depression (CSD) and peri-infarct depolarisation (PID) are related phenomena that have been associated with the human clinical syndromes of migraine (CSD), head injury and stroke (PID). Nevertheless the existence of CSD in man remains controversial, despite the detection of this phenomenon in the brains of most, if not all, other animal species investigated. This failure to unambiguously detect CSD clinically may be at least partly due to the anatomically complex, gyrencephalic structure of the human brain. This study was designed to establish conditions for the study of CSD in the brain of a gyrencephalic species using the noninvasive technique of magnetic resonance imaging (MRI). The 3-dimensional (3D) gyrencephalic anatomy of the cat brain was examined to determine the imaging conditions necessary to detect CSD events. Orthogonal transverse, sagittal and horizontal T1-weighted image slices showed that the marginal and suprasylvian gyri were the most appropriate cortical structures to study CSD. This was in view of (1) their simple geometry: (2) their lengthy extent of grey matter orientated rostrocaudally in the cortex: (3) their separation by a sulcus across which CSD spread could be studied and (4) the discontinuity in the grey matter in these regions between the right and left hemispheres dorsal to the corpus callosum. The structure suggested by the T1 weighted images was corroborated by systematic diffusion tensor imaging to map the fractional anisotropy and diffusion trace. Thus a single horizontal image plane could visualise the neighbouring suprasylvian and marginal gyri of both cerebral hemispheres, whereas its complex shape and position ruled out the ectosylvian gyrus for CSD studies. With the horizontal imaging plane, CSD events were reproducibly detected by animating successive diffusion-weighted MR images following local KCl stimulation of the cortical surface. In single image frames, CSD detection and characterisation required image subtraction or statistical mapping methods that, nevertheless, yielded concordant results. In repeat experiments, CSD events were qualitatively similar in appearance whether elicited by sustained or transient KCl applications. Our experimental approach thus successfully describes cat brain anatomy in vivo, and elucidates the necessary conditions for the application of MRI methods to detect CSD propagation. PMID- 11430694 TI - Differences between human and mouse alpha-fetoprotein expression during early development. AB - Alpha-fetoprotein (AFP) is the major serum protein during development. AFP is one of the earliest proteins to be synthesised by the embryonic liver. The synthesis of AFP decreases dramatically after birth and only trace amounts are expressed in the adult liver. The tissue distribution of AFP in early human embryogenesis has not been defined. We have studied the expression pattern of AFP mRNA in human and mouse embryos by in situ hybridisation. In humans, AFP is expressed in the hepatic diverticulum at 26 d postovulation as it differentiates from the foregut endoderm (i.e. in the most primitive hepatocytes). It is also expressed in the endoderm of the gastrointestinal tract and in the yolk sac at this age. AFP is subsequently expressed in the mesonephros and transiently in the developing pancreas. In the mouse, no expression of AFP was observed in the mesonephros but other sites of expression were similar. Thus AFP has a distinct temporospatial expression pattern during the embryonic period and this differs between human and mouse species. It is interesting that AFP is expressed by tumours such as primitive gastrointestinal, renal cell and pancreatic tumours as well as those of hepatocyte origin. This distribution reflects the sites of AFP expression during development. PMID- 11430695 TI - Retinoic acid specifically downregulates Fgf4 and inhibits posterior cell proliferation in the developing mouse autopod. AB - Retinoic acid, when administered to pregnant mice on d 11.0 of gestation, causes limb skeletal abnormalities consisting of reduced digital number, shortening of the long bones and delayed ossification. We show here that these effects are correlated with a decrease in cell proliferation within 5 h of retinoic acid administration, specifically in the posterior half of the distal limb bud mesenchyme, from which the distal skeletal elements are generated. There is a specific downregulation of Fgf4, a gene known to be involved in limb bud outgrowth and expressed only in the posterior part of the apical ectodermal ridge; Fgf8, which is expressed throughout the apical ectodermal ridge, is unaffected. The reduction in Fgf4 expression is not accompanied by downregulation of Shh, nor of its receptor and downstream target gene Ptc, suggesting that the skeletal reduction defects induced by retinoic acid are mediated specifically by FGF4-induced skeletogenic mesenchymal cell proliferation. PMID- 11430696 TI - Immunohistochemical identification of cells expressing ATP-gated cation channels (P2X receptors) in the adult rat thyroid. AB - We carried out immunohistochemistry and western blotting of fresh frozen sections and crude extracts from adult rat thyroids. The histochemical and immunoblotting studies were performed with P2X receptor antibodies from 2 different sources. P2X immunopositive cells were identified by fluorescence double labelling and confocal microscopy. Results of the western blotting experiments showed double bands of approximately 70 kDa and 140 kDa for all 7 P2X receptor subtypes with both sets of antibodies. Histochemical stains with antibodies from both sources also gave essentially identical results. P2X1, P2X, and P2X6 receptors were detected exclusively in vascular smooth muscle; P2X5 and P2X7 receptors were also present on vascular smooth muscle. Endothelial cells stained for P2X3, P2X4 and P2X7 receptors. Thyroid follicular cells displayed immunoreactivity for P2X3, P2X4 and P2X5 receptors. No immunostaining for P2X receptors was observed on C cells. Possible roles for the broad expression of P2X receptor subtypes in the rat thyroid are discussed. PMID- 11430697 TI - Immunolocalisation of the janus kinases (JAK)--signal transducers and activators of transcription (STAT) pathway in human epidermis. AB - The janus kinases (JAK) and signal transducers and activators of the transcription (STAT) pathway have been shown to be activated by a number of cytokines or growth factors and to play significant roles in the differentiation of various cell types. In the present study, we investigated the distribution of the JAK-STAT pathway using immunohistochemistry in the human epidermis. Each element of the pathway showed abundant and differential expression in the epidermis. The differential distribution of the elements was most strikingly observed in the horny keratinised cell and granular layers of the epidermis. JAK2, JAK3, STAT1 and STAT5 were expressed in high amounts, and JAK1, TYK2, STAT2, STAT3, STAT4 and STAT6 to a much lesser extent in the horny cell layer. JAK3, TYK2, STAT2, STAT3, STAT4 and STAT6 were more abundantly expressed in the granular layer than the lower layers of the epidermis. JAK1, STAT1 and STAT5 were expressed at almost the same levels in the various layers of the epidermis. These results show that elements of the JAK-STAT pathway are abundantly and differentially expressed in the epidermis. It is suggested that each element of the pathway may play a role at a distinct stage of keratinocyte differentiation. PMID- 11430698 TI - The three-dimensional feto-maternal vascular interrelationship during early bovine placental development: a scanning electron microscopical study. AB - Both the fetal and maternal microvasculature of bovine placentomes was examined by scanning electron microscopy of vascular casts. So far the development of the vascular architecture of the bovine placentome in early gestation has only been studied 2-dimensionally due to technical difficulties arising from the fragility of the early placental blood vessels. Repeated experiments led to the selection of the microvascular corrosion casts presented here. The vasculature of the maternal compartment is supplied by large caruncular stalk or spiral arteries, which release short maternal stem arteries. In the 3rd month of gestation, these arteries branch into several arterioles at their base, thus providing the vascular framework for the lower part of the septal walls of the primary crypts. In the 4th month, due to progressive longitudinal growth of the stem arteries, branching into arterioles occurs not only at the base, but over the whole length of the stem arteries. These arterioles supply the capillary complexes of the septa which resemble the major part of the septal vasculature and face the secondary crypts. Further indentation results in the formation of tertiary crypt capillary complexes, encircling the earlier secondary unit. From the 6th month of gestation the architecture resembles the fully developed maternal placenta with stem arteries running directly to the fetal side to branch into 4 to 6 arterioles, which turn back to enter secondary and tertiary septa. Maternal venules, collecting the blood from the capillary bed of secondary and tertiary septa, converge onto stem veins leaving the caruncle via branches of the uterine vein. The fetal part of the placentome is supplied by the cotyledonary arteries, which branch into fetal stem arteries that are the tributary to single villous trees. Over their whole course towards the maternal side, these give off arterioles entering secondary villi. The tertiary or terminal villous vasculature consists of capillaries, which are organised in serial capillary loops. This system is progressively elaborated in the course of gestation. In the 4th month there are only finger-like loops, whereas from the 6th month large fan-like structures can be observed. In early gestation the maternal and fetal blood vessels meet predominantly in a countercurrent fashion, changing to the less efficient crosscurrent exchange when the tertiary unit develops. These results indicate the development of a highly elaborated fetomaternal villous-crypt exchange system, already established in the 1st half of gestation, thus meeting the increasing needs of the fetus. PMID- 11430699 TI - Liver transplantation in man: morphometric analysis of the parenchymal alterations following cold ischaemia and warm ischaemia/reperfusion. AB - Ischaemia and reperfusion phases represent critical events during liver transplantation. The purpose of this study was to describe morphological alterations of both vascular and parenchymal compartments after ischaemia and reperfusion and to evaluate the possible relationship between morphometric parameters and biochemical/clinical data. Three needle biopsies were drawn from 20 patients who underwent orthotopic liver transplantation. The first biopsy was taken before flushing with preservation solution, and the second and the third to evaluate respectively the effects of cold ischaemia and of warm ischaemia/reperfusion. Biopsies were examined by an image analyser and morphometric parameters related to the liver parenchyma were evaluated. At the second biopsy we observed a decrease of the endothelium volume fraction while the same parameter referred to the sinusoidal lumen achieved a peak value. The hepatocytes showed a lower surface parenchymal/vascular sides ratio. This parameter was reversed at the end of the reperfusion phase; furthermore the third biopsy revealed endothelial swelling and a decreased volume fraction of the sinusoidal lumen. The results quantify the damage to the sinusoidal bed which, as already known, is one of the main targets of cold ischaemia; warm ischaemia and reperfusion accentuate endothelial damage. The end of transplantation is characterised by damage chiefly to parenchymal cells. Hepatocytes show a rearrangement of their surface sides, probably related to the alterations of the sinusoidal bed. In addition, the fluctuations of morphometric parameters during ischaemia/reperfusion correlate positively with biochemical data and clinical course of the patients. PMID- 11430700 TI - Ultrastructural response of pulmonary intravascular macrophages to exogenous oestrogen in the bovine lung: translocation of the surface-coat and enhanced cell membrane plasticity and angiogenesis. AB - The pulmonary intravascular macrophages (PIMs) of domestic ungulates are recognised by their specific surface coat, consisting of linearly arranged globules along the external leaf of the plasma membrane. The coat is sensitive to in vitro digestion with lipolytic lipase (LPL), intravenous heparin and clinical exposure to halothane anaesthesia. The sensitivity to these experimental manipulations suggests that the globules of the coat are predominantly composed of lipoproteins (LDL). The present administration of oestradiol proprionate in castrated male calves potentiated the translocation of the surface coat into the endocytotic pathway of the PIMs. Concurrently with mobilisation of the coat, the plasma membrane was thrown into prominent arrays of lamellipodial extensions. The sprawling macrophages made extensive adhesive contacts with the lining endothelium of the capillaries. Consequently, the endothelial cells were highly attenuated and precariously maintained the integrity of the vascular wall. At some focal points, the vascular wall was penetrated by the filopodial processes of PIMs, which protruded into the perivascular space. Furthermore, there were signs of neovascularisation in the form of overt mitotic changes, sprouting and precursor capillary formation. It is conceivable that the evolving profile of angiogenesis is due to the vascular endothelial growth factor (VEGF) paracrine function of PIMs. Endothelial cell specificity has been considered an important advantage of VEGF for neovascularisation. It allows pleotrophic response of endothelial cells to proliferate and to assemble into endothelial tubes. PMID- 11430701 TI - A novel branching pattern of the superior mesenteric artery found in the bullfrog (Rana catesbeiana) amphibian. AB - The branching and distribution patterns of the superior mesenteric artery were studied in 10 adult bullfrogs (Rana catesbeiana) after injection of coloured latex solution into the vasculature. The abdominal digestive organs in the bullfrog were mainly supplied by the coeliac artery and the superior mesenteric artery, both of which arose as a common trunk, the coeliacomesenteric artery, from the abdominal aorta. The coeliac artery supplied the stomach, liver, gallbladder and the pancreas, whereas the first branch of the superior mesenteric artery was the splenic artery with other branches supplying the greater part of intestine. The apex of the intestinal loop was defined as the region supplied by the trunk of the superior mesenteric artery, and its intestinal branches constituted a 'nested formation' which had the following characteristics. (1) The branches of the trunk were distributed to both sides of the apex, and the distribution regions of younger branches were located more distant from the apex than those of older branches. (2) Two branches directed towards both sides of the trunk frequently made a common stem arising from the trunk. The second branch of the superior mesenteric artery constituted a secondary trunk and its distribution region could be defined as a secondary apex, since 1 of its branches also constituted a nested formation which was distributed to both sides of the primary and secondary apices. The intestinal branches of the superior mesenteric artery were divided into 4 types on the basis of their pattern of branching and course. It is suggested that the nested formation of the superior mesenteric artery in the bullfrog is a remnant of the vascular pattern of the tadpole, which possesses a double spiral mode of intestinal convolution, probably supplied by arteries with the nested formation in a latent form. PMID- 11430702 TI - An alternative method for restoring opposition after median nerve injury: an anatomical feasibility study for the use of neurotisation. AB - Opposition, one of the most important functions of the hand, is lost or impaired after median nerve injury. Complete recovery does not always occur after treatment, and various techniques of opponensplasty are used for restoring opposition. This study was performed in order to develop an alternative method for selective restoration of thenar muscle function. Ten arms from 5 cadavers were used. The median nerve with its thenar motor branch (Tb) and the anterior interosseous nerve with its motor branch to pronator quadratus (PQb) were prepared in the distal forearm. The mean widths and the number of myelinated fibres of these nerves were: PQb 1.3+/-0.10 mm, Tb 1.4+/-0.12 mm and PQb 912+/-88 mm, Tb 1020+/-93 mm. The minimum necessary distance from the distal flexor crease of the wrist for neurotisation of the Tb by the PQb was 60+/-5.41 mm. It was concluded that PQb-Tb neurotisation would be possible anatomically. The advantages are that motor function is reestablished with a motor nerve, the diameters and the number of myelinated fibres of both nerves are similar, the loss of function after denervation of the pronator quadratus is slight and opponensplasty still remains as a final option. PMID- 11430703 TI - Prevention of pneumococcal disease in children. Pneumococcal conjugate vaccines: their use globally could have a major impact on public health. AB - Pneumococcal disease is a major cause of morbidity and mortality in infants and young children worldwide. New pneumococcal conjugate vaccines include 7 to 11 serotypes, which are the most common cause of paediatric disease in most parts of the world. The efficacy of a 7-valent conjugate vaccine was 97.4% (95% CI, 82.7 99.9) against invasive pneumococcal disease, and 57% (95% CI, 44-67) against otitis media, caused by vaccine serotypes. Evidence shows that the vaccine has the potential to prevent pneumonia. Pneumococcal conjugate vaccination has also been shown to reduce nasopharyngeal carriage of vaccine serotypes (particularly serotypes associated with antibiotic resistance). Thus widespread use of pneumococcal conjugate vaccine could substantially reduce the burden of invasive disease and would have the potential to control the global spread of antibiotic resistance in pneumococci. CONCLUSION: It is important that these highly effective vaccines should be made available to children in the developing countries. PMID- 11430704 TI - Introduction of solids and formula to breastfed infants: a longitudinal prospective study in Uppsala, Sweden. AB - The introduction of solids and formula was studied among 506 breastfed infants in Uppsala, Sweden, based on daily recordings during the first year. The mothers had previously breastfed at least 1 infant for at least 4 mo. Thirty-four per cent of the infants were introduced to solids before the age of 4 mo (4-6 mo is recommended in Sweden). Accustoming the infants to solids was a lengthy process. Life-table analyses showed a median duration of 28 d from the first introduction of solids to consumption of >10 ml daily, and 46 d before the infants ate > or = 100 ml of solids in 1 d for the first time. These durations were longer the younger the infant was at the introduction of solids. Thirty-two per cent of infants given formula consumed > or = 100 ml the first time it was given, and 49% did so within I wk, regardless of infant age. CONCLUSION: Parents and healthcare personnel need to be aware that accustoming breastfed infants to solid food is a lengthy process, and that there is a strong age effect on this duration. It is also important to consider what consequences the (usually) more abrupt introduction of formula might have on breastfeeding. PMID- 11430705 TI - Children's nutrient intake at preschool and at home. AB - A preschool-based dietary survey, using 7-d records, was carried out in a suburb of Stockholm. The aim was to assess the intake of food and the quality of the diet of preschool children aged 3-5 y at preschool and at home, and to compare the dietary intake with the Swedish dietary recommendations for preschool children. The respective mean intakes of protein, fat, carbohydrates and sucrose, expressed as a percentage of total energy intake were 14, 38, 50 and 9 at preschool, and at home 14, 36, 52 and 12 on weekdays, 14, 34, 55 and 16 on weekend days. The mean intakes of seven vitamins and minerals were low only for selenium as compared with the recommended level. No differences were found in nutrient density between diet at preschool and diet at home, with the exception of dietary fibre (higher at preschool). On weekdays there was a significantly higher nutrient density for calcium, zinc, selenium, vitamin A, riboflavin, vitamin C and dietary fibre compared with weekend days. CONCLUSION: The average intakes of energy and nutrients per meal at preschool compared with the recommended levels for children aged 4-6 y were low for all meals (breakfast, lunch and afternoon snack). This, however, was compensated for by home meals. PMID- 11430706 TI - Prevalence of iron deficiency in 12-mo-old infants from 11 European areas and influence of dietary factors on iron status (Euro-Growth study). AB - A prospective longitudinal cohort study was performed to assess the prevalence of iron deficiency in European infants at 12 mo of age, and to study the influence of socio-economic status, dietary factors, growth and morbidity on iron status. The cohort consisted of 488 normal term infants from primary healthcare centres in 11 European areas. Assessed were socio-economic variables, dietary intake, anthropometry and morbidity at regular intervals from birth to 12 mo, and haemoglobin, serum ferritin, mean corpuscular volume, transferrin saturation and serum transferrin receptor concentrations at age 12 mo. The prevalence of anaemia was 9.4%, of iron deficiency 7.2%, and of iron deficiency anaemia 2.3%. More than 40% of anaemia was associated with normal iron status and associated with an increased frequency of recent infections. Iron deficiency anaemia was significantly more frequent with low (5.1%) than high socio-economic status (0%). Dietary factors accounted for most of this variation in multiple regression analysis. Early introduction of cows' milk was the strongest negative determinant of iron status. Feeding of iron-fortified formula was the main factor positively influencing iron status. Other dietary factors, including breastfeeding, did not play a significant role as determinants of iron status at age 12 mo. Conclusion. Iron deficiency anaemia is present in 2.3% of 12-mo-old European infants. The prevalence of iron deficiency anaemia varies strongly with socio-economic status. Avoidance of cows' milk feeding during the first year of life is the key measure in the prevention of iron deficiency. PMID- 11430707 TI - Intestinal permeability in children: variation with age and reliability in the diagnosis of cow's milk allergy. AB - OBJECTIVE: To analyse to what extent age may alter intestinal permeability (IP) in children and to assess its reliability according to clinical manifestations in cow's milk allergy (CMA). DESIGN: A routine prospective study was performed in 200 children (12.5+/-23 mo, 0.5-168 mo) presenting with clinical manifestations suggesting CMA. Controls (n = 105) were those with a negative cow's milk challenge, whereas CMA children (n = 95) fulfilled ESPGHAN criteria. Permeability was measured as a percentage of urinary excretion of lactitol (L, %) and mannitol (M, %) (0.1 g/kg for each, oral absorption after a 6 h fast, 5 h urine collection, analysis by gas chromatography) and determination of the L/M ratio (L/M, %). RESULTS: In control children, L/M correlated negatively with age (r 0.33, p = 0.0006), whereas in those with CMA no correlation was found. Median L/M was significantly higher in CMA children (n = 95) than in controls (n = 105), 4.35+/-7.57% (95% CI 5.30-8.39%) vs 1.97+/-0.87% (95% CI 1.76-2.09%), (p = 0.0001). With a L/M cut-off value defined as mean + 2 SD of controls, in CMA IP exhibited a 68% sensitivity and a 77% negative predictive value. The highest sensitivity (70%) was seen at ages 6-12 mo and the lowest (58%) at age more than 3 y. An abnormal IP was seen in 80% of CMA children with digestive manifestations, in 43% with extra-digestive, 68% with mixed and 40% with anaphylactic manifestations. CONCLUSION: IP correlates negatively with age in control children and is altered in children with CMA. The test is at its most accurate in the diagnosis of CMA when done at ages 6-12 mo, when there are digestive manifestations. PMID- 11430708 TI - Efficacy of a packaged rice oral rehydration solution among children with cholera and cholera-like illness. AB - In past studies, patients with cholera and cholera-like diarrhoea treated with rice oral rehydration solution (ORS) had lower purging rates and a shorter illness duration. We evaluated a new packet form of rice ORS (CeraLyte-90) in 167 boys aged 5 to 15 y, with acute, dehydrating cholera and cholera-like diarrhoea in Bangladesh. The patients were randomized to receive either CeraLyte-90 (n = 85) or glucose ORS (n = 82) and were given early feeding and early antibiotics. The efficacy of the two solutions was compared for stool output during the first 8 h, the first 24 h, and total output, duration of diarrhoea, hematocrit, serum electrolytes and requirement for unscheduled intravenous fluids. The clinical and laboratory characteristics of the two groups were comparable on admission, and most of the patients had cholera (88% and 84% in the CeraLyte and glucose groups, respectively). The mean (+/- SE) stool output was 20% less in the rice ORS group during the first 8 h of treatment (86.2+/-6.6 ml/Kg vs 108.8+/-7.9 ml/Kg, p < 0.05), but the outputs during the other time periods were similar in the two groups, although children in the rice ORS group had slightly more vomiting on day one (p < 0.05). The mean serum electrolyte concentrations in both groups of children remained within normal range. CONCLUSION: The study documents the safety and efficacy of the new, packaged rice ORS. PMID- 11430709 TI - Fluid resuscitation in infantile hypertrophic pyloric stenosis. AB - The purpose of this analysis was to investigate biochemical disturbances at presentation and initial fluid resuscitation before surgery in infantile pyloric stenosis. The charts of 139 consecutive infants (113 boys and 26 girls) between 7 d and 20 wk of age with hypertrophic pyloric stenosis were reviewed. The infants were treated at the Department of Pediatric Surgery, University of Bern, Switzerland, in the period between 1987 and 1997. A trend towards hypokalaemia (13 of the 139 patients), hypochloraemia (39 patients) and especially metabolic alkalosis (98 patients) was frequently noted on admission. In 84 patients, data on fluid management and on circulating sodium, potassium, chloride and the acid base balance immediately before surgery were also available. In these patients a significant correlation was found between the parenteral chloride dose given for fluid repair (y = 0.310 x; r = 0.54; p < 0.001) and the changes in plasma bicarbonate. The equation indicates that a chloride dose of 10 mmol/kg body weight is required to reduce plasma bicarbonate on average by 3 mmol/. CONCLUSION: Since assessment of the fluid volume stated by physical examination and history is inaccurate in infants with vomiting, the severity of metabolic alkalosis helps to define the amount of fluid required for repair. PMID- 11430710 TI - Insulin improves clinical status of patients with cystic-fibrosis-related diabetes mellitus. AB - Cystic-fibrosis-related diabetes mellitus is frequently underdiagnosed and associated with deterioration of overall clinical status. The purpose of this prospective study was to investigate the influence of insulin on nutrition, lung function and clinical status of cystic fibrosis patients. For a period of 5 y, and at 6-mo intervals, body mass index, forced expiratory volume in 1 sec, Shwachman score, intravenous glucose tolerance test and first-phase insulin response were determined in 30 cystic fibrosis patients (age range 10-35 y) with exocrine pancreatic insufficiency. During the study period, six patients (3M and 3F; age range 15-22 y) developed diabetes and required insulin therapy. The decrease of first-phase insulin response coincided with deterioration of nutritional and clinical status, which improved significantly 6 mo after the institution of insulin. CONCLUSION: Insulin, as an anabolic hormone, could have an influence on body mass, which may affect pulmonary function and clinical condition in cystic fibrosis. It is important to identify cystic fibrosis individuals at risk of developing diabetes so that early insulin therapy is instituted. PMID- 11430711 TI - Effects of growth hormone treatment on the leptin system and body composition in obese prepubertal boys. AB - Leptin correlates with measures of body fat stores. Growth hormone (GH) treatment may affect leptin levels either directly or indirectly by influencing body composition and circulating insulin level. Here, the effects of GH treatment on the leptin axis and body composition of six severely obese, but otherwise healthy, prepubertal boys were studied. Fasting serum leptin was significantly reduced after only 3 wk of GH treatment. Body fat percentage, but not BMI, decreased significantly (p < 0.05) after 3 mo. The serum leptin concentration per unit fat mass decreased significantly during GH treatment (p < 0.05), suggesting that such treatment might have a direct effect on serum leptin independently of the effects on body composition. Leptin RNA expression in abdominal subcutaneous tissue was not significantly changed by treatment. CONCLUSIONS: The data indicate that GH has an early downregulatory effect on the circulating leptin level independently of the concomitant changes in body composition. Whether GH affects leptin production or metabolism needs further study. PMID- 11430712 TI - Morning versus evening administration of estradiol to girls with turner syndrome receiving growth hormone: impact on growth hormone and metabolism. A randomized placebo-controlled crossover study. AB - Timing of 17beta-estradiol (E2) administration in relation to that of GH could influence the "first pass effect" of E2 on hepatic IGF-I secretion. In order to test this hypothesis, a randomized double-blind placebo-controlled crossover study was conducted. Nine Turner girls (12.8-20.0y) were treated for 2 mo periods with GH 0.1 IU/kg/d sc at bedtime, and oral E2 6-11 microg/kg/d in the morning and placebo in the evening in one 2-mo period and vice versa in the other period. After each period, 24-h blood sampling was performed. IGF-I and mean 24-h integrated GH were comparable. However, the IGF-I/IGFBP-3 ratio was higher (p = 0.05) and insulin levels were lower after evening administration of E2 (24 h: p = 0.03). During an oral glucose tolerance test in the morning, glucagon and insulin were lower following evening E2 administration (ANOVA: glucagon, p = 0.03; insulin, p = 0.04), as well as insulin resistance tended to be lower (p = 0.09). CONCLUSION: The timing of oral E2 supplementation modulates the IGF-I/IGFBP-3 ratio, insulin and glucagon levels in Turner syndrome during GH treatment, Evening administration of oral estrogen together with evening injections of GH may be preferable. PMID- 11430713 TI - Pre-school children with developmental co-ordination disorder: self-perceived competence and group motor skill intervention. AB - OBJECTIVE: To investigate self-perceived competence in 5- to 6-y-old children with developmental co-ordination disorder. SUBJECTS AND METHODS: Scores in two scales of self-perceived competence were compared between a study group of children with developmental co-ordination disorder (n = 37) and a reference group (n = 60). In addition, effects of group motor skill intervention were evaluated by comparing scores in study group children randomly allocated to an intervention (n = 17) and a control subgroup (n = 20). The intervention subgroup received intervention once weekly for 10 wk. CONCLUSIONS: Pre-school children with developmental co-ordination disorder are similar to peers of the same age regarding self-perception of their own competence in areas concerning school, peer relations and motor skills. The change in score in the perceived motor competence scale was significantly greater in the intervention than in the control subgroup, suggesting that the intervention increased individual awareness of motor competence. PMID- 11430714 TI - Development and behaviour of seven-year-old triplets. AB - The aim of this study was to assess the development and behaviour of 7-y-old triplets, who were followed from delivery to 7 y of age. The McCarthy Scales of Children's Abilities (MSCA) and Achenbach's Child Behaviour Checklist (CBCL) were used to assess and compare development and behaviour in the triplets and in a control group of singletons. The sample included 33 sets of triplets and 33 singletons, all 7-y-olds, and comparable for sex and fathers' social class. The triplets' scores on the MSCA were all within normal range. Although their mean scores for all subtests were below those of singletons, they did not differ significantly. The mean CBCL scores, based on mothers' reports, indicated that the triplets had significantly fewer behavioural problems than the controls. In half the triplet sets, one child stood out as having more behavioural problems than the others. CONCLUSION: Further assessments of the development of triplets are needed; these should be based on larger study samples and should use information from teachers and external observers as well as mothers. PMID- 11430715 TI - Premature cessation of breastfeeding in infants: development and evaluation of a predictive model in two Argentinian cohorts: the CLACYD study, 1993-1999. Cordoba Lactation, Feeding, Growth and Development study. AB - The objective of this study was to develop a model to predict premature cessation of breastfeeding of newborns, in order to detect at-risk groups that would benefit from special assistance programmes. The model was constructed using 700 children with a birthweight of 2000 g or more, in 2 representative cohorts in 1993 and 1995 (CLACYD I sample) in Cordoba, Argentina. Data were analysed from 632 of the cases. Mothers were selected during hospital admittance for childbirth and interviewed in their homes at 1 mo and 6 mo. To evaluate the model, an additional sample with similar characteristics was drawn during 1998 (CLACYD II sample). A questionnaire was administered to 347 mothers during the first 24-48 h after birth and a follow-up was completed at 6 mo, with weaning information on 291 cases. Premature cessation of breastfeeding was considered when it occurred prior to 6 mo. A logistic regression model was fitted to predict premature end of breastfeeding, and was applied to the CLACYD II sample. The calibration (Hosmer Lemeshow C statistic) and the discrimination [area under the receiver operating characteristics (ROC) curve] of the model were evaluated. The predictive factors of premature end of breastfeeding were: mother breastfed for less than 6 mo [odds ratio (OR) = 1.84, 95% confidence interval (CI) 1.26-2.70], breastfeeding of previous child for less than 6 mo (OR = 4.01, 95% CI 2.58-6.20), the condition of the firstborn child (OR = 2.75, 95% CI 1.79-4.21), the first mother-child contact occurring after 90 min of life (OR = 1.88; 95% CI 1.22-2.91) and having an unplanned pregnancy (OR = 1.50, 95% CI 1.05-2.15). The calibration of the model was acceptable in the CLACYD I sample (p = 0.54), as well as in the CLACYD II sample (p = 0.18). The areas under the ROC curve were 0.72 and 0.68, respectively. CONCLUSION: A model has been suggested that provides some insight onto background factors for the premature end of breastfeeding. Although some limitations prevent its general use at a population level, it may be a useful tool in the identification of women with a high probability of early weaning. PMID- 11430716 TI - Subjective daytime sleepiness and its predictors in Finnish adolescents in an interview study. AB - The purpose of this study is to evaluate the predictors of subjective daytime sleepiness (SDS) and its chronicity in adolescents. Two groups of adolescents (107 with SDS and 107 without SDS) from our first questionnaire study were invited to an interview after 3 y. A follow-up questionnaire had been sent to them one year earlier. The interview included questions about sleep, daytime sleepiness, living habits, physical and mental health, and progress at school. The adolescents were also examined clinically. Interviews were conducted with 66 out of 107 subjects with SDS and 64 out of 107 without SDS (age range 12 to 19 y). In this interview 42 out of the 130 adolescents had SDS. A total of 20 adolescents reported SDS in both questionnaire studies and in the interview (chronic SDS). In a multivariate analysis (logistic regression) sleep disorders, frequent medication and depressive emotions were significantly associated with SDS. Chronic SDS was connected in a bivariate analysis (Pearson's chi-square) with excessive night waking, difficulty in falling asleep, dreaming, frequent medication, frequent alcohol drinking, and irregular breakfast eating, and in our previous studies also with delayed sleep rhythm. CONCLUSION: Sleep disorders and health problems were more common causes of SDS than undesirable living habits. However, alcohol drinking and delayed sleep rhythm were associated with chronic SDS in addition to sleep disorders and medication. PMID- 11430717 TI - Person, time and place coordinates of pedestrian injuries: a study in Athens. AB - The aim of this study was to investigate whether socioeconomic status of town of residence is associated with risk for childhood pedestrian injuries. The study population consisted of all pedestrian victims, aged 0-14 y, who lived in towns of Greater Athens and who presented to the Emergency Department of a major Children's Hospital during the period 1996-98. The towns were divided into three categories by socioeconomic status according to the proportion of (a) adult household heads with a higher education degree and (b) households with less than one person per room. The rate of pedestrian injuries was estimated by socioeconomic status of the residential town and by place of accident (inside or outside the respective town). The pedestrian injury rate ranged from 5.5 to 12 injured children among a 10000 childhood population per year, with an almost twofold excess among children residing in the less wealthy towns compared with the wealthier ones. The social gradient was steeper for injuries occurring outside the residential town. The population fractions of pedestrian injury rates attributable to educational level and household crowding differentials, regardless of the place of accident, were 39% and 25%, respectively. CONCLUSIONS: There is a considerable social gradient for childhood injuries irrespective of place of accident, a finding that could be partly attributable to lower socioeconomic background rather than to adverse environmental factors prevailing in less wealthy towns. Our findings indicate that there is a need for preventive programmes targeting people as well as places of low socioeconomic status. PMID- 11430718 TI - Traumatic experiences and post-traumatic stress reactions in children from Kurdistan and Sweden. AB - This paper examines the types and scores of traumatic experiences, post-traumatic stress symptom and behavioural disorders among Kurdistanian refugee children in Sweden and a comparative Swedish group. The Harvard-Uppsala Trauma Questionnaire for Children (HUTQ-C), the Post-Traumatic Stress Disorder for Children (PTSS-C) and the Child Behaviour Checklist (CBCL) were administered in interview form to 32 children from each sample, controlled for age, gender and trauma levels. No significant differences were found between the 2 samples regarding types of traumatic events, frequencies of post-traumatic stress disorder, post-traumatic stress symptom scores or behavioural problem scores, except in 3 aspects: Kurdistanian children reported more war experience and being lost, while Swedish children presented higher frequencies of leisure-time accidents. CONCLUSION: This study supports the assumption that children differ from adults by showing more similarities than differences regarding traumatic experiences and post-traumatic stress reactions, after being exposed to an equivalent number of reported traumatic events. The refugee children's stay in Sweden can be considered as a healing factor. PMID- 11430719 TI - Nitric oxide production during the early neonatal period in small-for-gestational age infants. AB - In a study of endogenous nitric oxide production in growth-retarded, very preterm newborns (<32 wk GA), urinary NOx/creatinine ratio and plasma guanosine 3',5' cyclic monophosphate levels were determined during the early neonatal period. Newborns were divided into three groups: appropriate-for-gestational-age (AGA, n = 19), moderately small-for-gestational-age (SGA, n = 13) and severely SGA (n = 6) infants. Severely SGA infants showed significant higher values of nitric oxide derivatives during the first 24 h of life compared with the other groups. CONCLUSION: An increased NO production is found in SGA infants during the first 24 h after birth. This may reflect an increased intrauterine nitric oxide production in the feto-placental circulation found in cases with intrauterine growth retardation, PMID- 11430720 TI - Neurodevelopmental outcome in high-risk preterm infants treated with inhaled nitric oxide. AB - Inhaled nitric oxide (iNO) is used to treat preterm infants with hypoxaemic respiratory failure. In this study we describe the long-term survival and neurodevelopmental status of high-risk preterm infants enrolled into a randomized controlled trial of iNO therapy. Information regarding long-term outcome was available for all 25 children enrolled in the original trial who survived until discharge from hospital. Formal, blinded, developmental assessment and neurological examinations were performed in 21 out of 22 children still alive at 30 mo of age, corrected for prematurity. No significant differences were found in long-term mortality (12/20 vs 8/22, RR 1.65, 95% CI 0.87-3.3), neurodevelopmental delay (4/7 vs 9/14, RR 0.89, 95% CI 0.37-1.75), severe neurodisability (0/7 vs 5/14, p = 0.12) or cerebral palsy (0/7 vs 2/14, p = 0.53) between iNO-treated and control infants. CONCLUSION: In this study there was no evidence of a significant effect on either survival or long-term neurodevelopmental status in infants treated with iNO. PMID- 11430721 TI - Vitamin D deficiency in Iranian mothers and their neonates: a pilot study. AB - We conducted a pilot study to assess the prevalence of hypovitaminosis D among Iranian women and their newborns. Blood samples were taken from 50 mothers (age 16-40 yr) and their neonates at term delivery in the largest Tehran hospital. The results showed that 80% of the women had 25-hydroxyvitamin D (25-OHD) concentrations of less than 25 nmol/l. Mean maternal plasma calcium and phosphatase alkaline concentrations were in the normal range. The mean maternal serum immunoreactive parathyroid hormone concentration of women with hypovitaminosis D (i.e., 25-OHD levels <25 nmol/l) was above normal range and significantly different from that of women without hypovitaminosis D. CONCLUSION: The mean cord serum 25-OHD concentration was very low (4.94+/-9.4 nmol/l) and that of infants of mother with hypovitaminosis D were almost undetectable (1.2+/ 1.2 nmol/l). PMID- 11430722 TI - Microcephalus, medulloblastoma and excessive toxicity from chemotherapy: an unusual presentation of Fanconi anaemia. AB - Fanconi anaemia is a genetically and phenotypically heterogeneous disorder with different forms of clinical presentation. In this case the patient had suffered from microcephalus and delayed motor development from birth, but extensive investigation did not disclose any aetiology. At 3.5 y she developed a cerebellar medulloblastoma which was treated with surgery and chemotherapy. Following chemotherapy with alkylating agents she suffered from severe bone marrow aplasia which caused life-threatening infections, feeding problems and impaired kidney function. Fanconi anaemia was suspected, but it took 2 mo before the chromosome fragility test came out positive. From the moment diagnosis of Fanconi anaemia was made, no further active treatment was given. The patient's condition improved for some time, but she relapsed and died exactly 1 y after the first diagnosis of brain tumour. CONCLUSION: Fanconi anaemia must always be suspected in patients who experience excessive toxicity from chemotherapy regardless of the type of malignancy and congenital malformations. PMID- 11430723 TI - Non-Epstein-Barr virus-associated T-cell lymphoma following cardiac transplantation for Barth syndrome. AB - Barth syndrome is a metabolic disease characterized by infantile cardiomyopathy, neutropenia and organic aciduria. We report disease evolution in one of the first affected boys to undergo successful cardiac transplantation. CONCLUSION: Although cardiac status stabilized, he developed disabling skeletal myopathy, protracted lymphopenia and--5 y after transplant--fatal Epstein Barr (EBV)-negative T-cell non-Hodgkin's lymphoma. PMID- 11430724 TI - Torticollis revealing as a symptom of acute lymphoblastic leukaemia in a fourteen month-old girl. AB - The differential diagnosis of acquired torticollis in infants and children comprises a large variety of disorders, including inflammation, neurologic disease or trauma. CONCLUSION: Spinal cord tumours in patients with ALL are a rarely reported entity and secondary spinal involvement in leukaemia presenting as head tilt is even more uncommon. PMID- 11430725 TI - Continuing progressive deterioration of the environment in the Aral Sea Region: disastrous effects on mother and child health. AB - Scientists, non-governmental experts (NGOs) and governmental officials from the Central Asian Republics and an international group of invited scientists and NGO representatives participated in a workshop on the disastrous health problems in the Aral Sea Region. Various serious problems were reported in more than 20 presentations. Particular emphasis was put on the way in which adverse environmental factors such as contaminated water and food have contributed to the deterioration of human health, particularly that of mothers and children. CONCLUSION: There is an urgent request that the international community assists local scientists to develop programmes to improve the health of the population in the Aral Sea Region. PMID- 11430727 TI - Fluid balance, respiratory status and limiting the risks of postnatal dexamethasone. PMID- 11430728 TI - Familial mercury intoxication presenting with cardiovascular abnormalities and acrodynia. PMID- 11430729 TI - Mycoplasma pneumoniae infection associated with Kawasaki disease. PMID- 11430730 TI - Further evidence that fibroblast growth factor receptor 2 mutations cause Antley Bixler syndrome. PMID- 11430731 TI - Inclusion of osteopathic orthopedists in workforce studies. PMID- 11430732 TI - Safe administration of aprotinin in orthopedic surgery. PMID- 11430733 TI - Sports medicine update: where have we been, where are we going? PMID- 11430734 TI - Tibial nails for femoral shaft fractures in adolescents: "off-label" usage. PMID- 11430735 TI - Degenerative spondyloarthropathy of the cervical and lumbar spine in jockeys. AB - This prospective study analyzed the long-term effects of horse riding on the cervical and lumbar spine of jockeys. Thirty-two jockeys were observed for 13 years. All jockeys had clinical and radiographic evaluation of the spine; three consecutive age groups were studied. Results were compared to age-matched, normal population control groups. The incidence of degenerative changes of the spine was higher in the jockeys compared with the control groups and was more prominent in the older age group for both the lumbar and cervical spine. These findings suggest that equestrian sports, particularly professional horse riding, apart from the increased risk of direct spinal injury caused by a fall from the horse, can lead to progressive spine degeneration as a result of repetitive trauma and increased physical stress on the spine. PMID- 11430736 TI - Triple innominate osteotomy in young adults for the treatment of acetabular dysplasia: a 9-year follow-up study. AB - Fifty patients who underwent 60 triple innominate osteotomies were reviewed radiographically and clinically using a modified Harris hip score (HHS). Average patient age was 26 years (range: 13-48 years). At average 9-year follow-up (range: 5-14 years), 12 (20%) hips had been converted to total hip arthroplasty (THA) and 4 (7%) hips had incapacitating pain. Sixteen (27%) hips were considered failures. Average modified HHS at final follow-up was 67 (range: 28-91). Forty nine (98%) of 50 patients reported they would recommend the procedure to others in the same situation. Radiographically, there was significant improvement in the center-to-edge angle of Wiberg and the acetabular angle of Sharp. There also was a statistically significant relationship between failure of the osteotomy and severity of preexisting hip arthrosis as measured by the Tonnis criteria. The results demonstrate triple innominate osteotomy was effective in eliminating pain, but the fact that 27% of hips required or will require THA indicates results may deteriorate with time. PMID- 11430737 TI - Incidence of bilateral proximal femoral fractures in a tertiary care center. AB - Over a 7-year period, a total of 545 osteoporotic fractures involving the proximal femur were treated at our institution. Twenty-nine patients sustained fractures of both hips, representing an overall incidence of bilateral hip fractures of approximately 5.5%. Of the 29 patients with bilateral hip fractures, adequate records and radiographs were avaialable for 28. There was a significant similarity between fracture patterns in the majority of bilateral hip fracture patients: 18 (64%) patients demonstrated similar fracture patterns on both sides. In these 18 patients, the average interval between fractures was approximately 1.7 years; in the remaining 10 patients, in whom the fractures were of dissimilar nature, the average interval between fractures was approximately 3.5 years. These findings suggest that at least 1 in 20 patients who sustain a fracture about the hip can expect to suffer a fracture involving the contralateral side. Moreover, the difference in the average interval for fractures of similar nature compared to fractures of dissimilar nature suggests the effects of physiologic age and the accordant changes in bone architecture play a larger role in predicting subsequent fracture pattern. PMID- 11430738 TI - Preoperative duplex ultrasonography evaluation for deep venous thrombosis in revision hip arthroplasty patients. AB - In a prospective consecutive series, 53 revision hip arthroplasties were performed in 51 patients. Pre- and postoperative Duplex ultrasonography examinations were reviewed by an independent, experienced radiologist. Three of 51 patients (53 procedures) had evidence of chronic deep venous thrombosis (DVT) or other venous abnormality preoperatively, yielding an incidence of 5.6%. One (1.9%) patient developed an acute DVT postoperatively despite pharmacological and mechanical preventative measures. These results indicate the use of preoperative ultrasonography as a screening tool prior to revision hip arthroplasty is not warranted based on the low incidence of acute or chronic DVT detected preoperatively. Long-term anticoagulation, when necessary, can be based on the findings of a postoperative scan. PMID- 11430739 TI - Tosic external fixator in the management of proximal tibial fractures in adults. AB - This retrospective clinical study assessed proximal tibial fractures managed with the Tosic external fixator. Nineteen patients with 21 proximal tibial fractures treated with the Tosic external fixator between July 1997 and October 1998 comprised the study population. Eleven fractures were graded as 41A2, 3 fractures as 41 A3, 4 fractures as 41C1, and 3 fractures as 41 C2. Fourteen fractures were closed, and 7 fractures were open. Average time to healing was 1 7 weeks. No revision of fixation was needed. There were five cases of pin tract infection. Average range of knee motion was 2 degrees-135 degrees. These results indicate the Tosic external fixator is an efficient and simple way to treat proximal tibial metaphyseal fractures. PMID- 11430740 TI - Culture results and amputation rates in high-pressure paint gun injuries of the hand. AB - High-pressure paint gun injuries have been well described in the literature, and the use of antibiotics is recommended as part of their management. However, there is no scientific evidence to support the use of antibiotics. In addition, the type of paint injected (water- versus oil-based) has never been investigated to determine the extent of morbidity resulting from these injuries. This study examines the organisms cultured in wounds resulting from these injuries and whether the type of paint injected had an influence on amputation rates. Charts of 35 patients with high-pressure paint gun injuries to their hands were reviewed. The amputation rate was 50% with oil-based paints and 0% with water based paints. Forty-seven percent of wound cultures were positive, with gram negative bacteria found in 58% of isolates. Our findings support the use of antibiotics, which should cover both gram-positive and gram-negative organisms. PMID- 11430741 TI - Adolescent quadriceps mechanism disruption. PMID- 11430742 TI - Fracture-dislocation of the scapula. PMID- 11430743 TI - Nonunion of a dominant side first rib stress fracture in a baseball pitcher. PMID- 11430744 TI - Myositis ossificans and hip ankylosis secondary to a muscle strain injury. PMID- 11430745 TI - Lipoma arborescens of the knee as a possible cause of osteoarthrosis. PMID- 11430746 TI - Dermoid cyst of the sternum. PMID- 11430747 TI - Lyme arthritis in a child with HIV. PMID- 11430748 TI - Evaluation of the risk of pathologic fractures secondary to metastatic bone disease. PMID- 11430749 TI - Side chain mobility as monitored by CH-CH cross correlation: the example of cytochrome b5. AB - The mobility of betaCH2 moieties in oxidized and reduced cytochrome b5 was studied by analyzing the 13C relaxation of the J-split components, in terms of C H dipole-C-H dipole cross correlation rates. A 2D 13C-1H experiment is proposed to measure these rates that provide the internal effective reorientation correlation time for each CH2 moiety. It is found that higher mobility is present in the alpha helices forming the heme pocket. On the contrary, the beta strands, which form the hydrophobic core of the molecule, have the lowest mobility. The general pattern is the same for the oxidized and reduced species, indicating that any oxidation-dependent property detected for backbone NH moieties does not affect the CH2 mobility. PMID- 11430750 TI - A solubility-enhancement tag (SET) for NMR studies of poorly behaving proteins. AB - Protein-fusion constructs have been used with great success for enhancing expression of soluble recombinant protein and as tags for affinity purification. Unfortunately the most popular tags, such as GST and MBP, are large, which hinders direct NMR studies of the fusion proteins. Cleavage of the fusion proteins often re-introduces problems with solubility and stability. Here we describe the use of N-terminally fused protein G (B1 domain) as a non-cleavable solubility-enhancement tag (SET) for structure determination of a dimeric protein complex. The SET enhances the solubility and stability of the fusion product dramatically while not interacting directly with the protein of interest. This approach can be used for structural characterization of poorly behaving protein systems, and would be especially useful for structural genomics studies. PMID- 11430751 TI - Structure prediction of protein complexes by an NMR-based protein docking algorithm. AB - Protein docking algorithms can be used to study the driving forces and reaction mechanisms of docking processes. They are also able to speed up the lengthy process of experimental structure elucidation of protein complexes by proposing potential structures. In this paper, we are discussing a variant of the protein protein docking problem, where the input consists of the tertiary structures of proteins A and B plus an unassigned one-dimensional 1H-NMR spectrum of the complex AB. We present a new scoring function for evaluating and ranking potential complex structures produced by a docking algorithm. The scoring function computes a 'theoretical' 1H-NMR spectrum for each tentative complex structure and subtracts the calculated spectrum from the experimental one. The absolute areas of the difference spectra are then used to rank the potential complex structures. In contrast to formerly published approaches (e.g. [Morelli et al. (2000) Biochemistry, 39, 2530-2537]) we do not use distance constraints (intermolecular NOE constraints). We have tested the approach with four protein complexes whose three-dimensional structures are stored in the PDB data bank (Bernstein et al., 1977) and whose 1H-NMR shift assignments are available from the BMRB database. The best result was obtained for an example, where all standard scoring functions failed completely. Here, our new scoring function achieved an almost perfect separation between good approximations of the true complex structure and false positives. PMID- 11430752 TI - Measurement of homonuclear three-bond J(H(N)Halpha) coupling constants in unlabeled peptides complexed with labeled proteins: application to a decapeptide inhibitor bound to the proteinase domain of the NS3 protein of hepatitis C virus (HCV). AB - A new isotope-filtered experiment has been designed to measure homonuclear three bond J(H(N)Halpha) coupling constants of unlabeled peptides complexed with labeled proteins. The new experiment is based on the 3D HNHA pulse scheme, and belongs to the 'quantitative J-correlation' type. It has been applied to a decapeptide inhibitor bound to the proteinase domain of the NS3 protein of human hepatitis C virus (HCV). PMID- 11430753 TI - Geometry dependent two-dimensional heteronuclear multiplet effects in paramagnetic proteins. AB - We report experimental observation and numerical simulation of a two-dimensional multiplet effect in the heteronuclear correlation spectrum of a paramagnetic protein that depends on molecular geometry. This effect arises as a consequence of cross-correlated relaxation involving the Curie spin relaxation and internuclear dipolar relaxation mechanisms. It also manifests itself in resolution and sensitivity improvement in transverse relaxation optimised spectroscopy (TROSY) kind of experiments. Characteristic multiplet patterns in heteronuclear coupled two-dimensional NMR spectra encode directional information for the heteronuclear bond with respect to the paramagnetic center. These patterns, which are simulated here using Redfield's relaxation theory, can be used to obtain a new type of geometry restriction for structure determination and refinement of paramagnetic macromolecular systems. PMID- 11430754 TI - Measurement and application of 1H-19F dipolar couplings in the structure determination of 2'-fluorolabeled RNA. AB - Residual dipolar couplings can provide powerful restraints for determination and refinement of the solution structure of macromolecules. The application of these couplings in nucleic acid structure elucidation can have an especially dramatic impact, since they provide long-range restraints, typically absent in NOE and J coupling measurements. Here we describe sensitive X-filtered-E.COSY-type methods designed to measure both the sign and magnitude of long-range 1H-19F dipolar couplings in selectively fluorine labeled RNA oligonucleotides oriented in solution by a liquid crystalline medium. The techniques for measuring 1H-19F dipolar couplings are demonstrated on a 21-mer RNA hairpin, which has been specifically labeled with fluorine at the 2'-hydroxyl position of three ribose sugars. Experimentally measured 1H-19F dipolar couplings for the 2'-deoxy-2' fluoro-sugars located in the helical region of the RNA hairpin were found to be in excellent agreement with values predicted using canonical A-form helical geometry, demonstrating that these couplings can provide accurate restraints for the refinement of RNA structures determined by NMR. PMID- 11430755 TI - MUNIN: a new approach to multi-dimensional NMR spectra interpretation. AB - A new method, MUNIN (Multi-dimensional NMR spectra interpretation), is introduced for the automated interpretation of three-dimensional NMR spectra. It is based on a mathematical concept referred to as three-way decomposition. An NMR spectrum is decomposed into a sum of components, with each component corresponding to one or a group of peaks. Each component is defined as the direct product of three one dimensional shapes. A consequence is reduction in dimensionality of the spectral data used in further analysis. The decomposition may be applied to frequency domain or time-domain data, or to a mixture of these. Features of MUNIN include good resolution in crowded regions and the absence of assumptions about line shapes. Uniform sampling of time-domain data, a prerequisite for discrete Fourier transform, is not required. This opens an avenue for the processing of NMR data that do not follow oscillating behaviour, e.g. from relaxation measurements. The application of MUNIN is illustrated for a 1H-15N-NOESY-HSQC, where each component is defined as the set of all NOE peaks formed by a given amide group. As a result, the extraction of structural information simply consists of one dimensional peak picking of the shape along the NOE-axis obtained for each amide group. PMID- 11430756 TI - Principal components analysis of protein structure ensembles calculated using NMR data. AB - One important problem when calculating structures of biomolecules from NMR data is distinguishing converged structures from outlier structures. This paper describes how Principal Components Analysis (PCA) has the potential to classify calculated structures automatically, according to correlated structural variation across the population. PCA analysis has the additional advantage that it highlights regions of proteins which are varying across the population. To apply PCA, protein structures have to be reduced in complexity and this paper describes two different representations of protein structures which achieve this. The calculated structures of a 28 amino acid peptide are used to demonstrate the methods. The two different representations of protein structure are shown to give equivalent results, and correct results are obtained even though the ensemble of structures used as an example contains two different protein conformations. The PCA analysis also correctly identifies the structural differences between the two conformations. PMID- 11430757 TI - A method for efficient isotopic labeling of recombinant proteins. AB - A rapid and efficient approach for preparing isotopically labeled recombinant proteins is presented. The method is demonstrated for 13C labeling of the C terminal domain of angiopoietin-2, 15N labeling of ubiquitin and for 2H/13C/15N labeling of the Escherichia coli outer-membrane lipoprotein Lpp-56. The production method generates cell mass using unlabeled rich media followed by exchange into a small volume of labeled media at high cell density. Following a short period for growth recovery and unlabeled metabolite clearance, the cells are induced. The expression yields obtained provide a fourfold to eightfold reduction in isotope costs using simple shake flask growths. PMID- 11430758 TI - Multiplet component separation for measurement of methyl 13C-1H dipolar couplings in weakly aligned proteins. AB - A simple spectral editing procedure is described that generates separate subspectra for the methyl 13C-[1H3] multiplet components of 1H-13C HSQC spectra. The editing procedure relies on co-addition of in-phase and antiphase spectra and yields 1H-coupled constant-time HSQC subspectra for the methyl region that have the simplicity of the regular decoupled CT-HSQC spectrum. Resulting spectra permit rapid and reliable measurement of 1H-13C J and dipolar couplings. The editing procedure is illustrated for a Ca2+-calmodulin sample in isotropic and liquid crystalline phases. PMID- 11430759 TI - Domain orientation in beta-cyclodextrin-loaded maltose binding protein: diffusion anisotropy measurements confirm the results of a dipolar coupling study. AB - Maltose binding protein (MBP) is a 370-residue two-domain molecule involved in bacterial chemotaxis and sugar uptake. Rotational diffusion tensors were calculated for a complex between MBP and beta-cyclodextrin using backbone 15N T1 and T1rho relaxation times and steady state 1H-15N NOE values. The tensors obtained for each of the two domains in the protein were subsequently used to determine the relative domain orientation in the molecule. The average domain orientation determined using this approach agrees well with results from dipolar coupling data, but differs significantly from the domain orientation deduced from X-ray studies of the complex. PMID- 11430760 TI - 1H, 15N and 13C resonance assignments for the Tctex1 dynein light chain from Chlamydomonas flagella. PMID- 11430761 TI - Assignment of 1H, 15N and 13C resonances of the carbohydrate recognition domain of human galectin-3. PMID- 11430762 TI - 1H, 13C and 15N resonance assignments and secondary structure of the cytotoxic protein RNase 4 from bullfrog Rana catesbeiana oocytes. PMID- 11430763 TI - 1H, 13C and 15N backbone resonance assignment of the arsenate reductase from Staphylococcus aureus in its reduced state. PMID- 11430764 TI - Backbone H(N), N, Calpha, C' and Cbeta assignment of the 25 kDa peptide methionine sulfoxide reductase from Erwinia chrysanthemi. PMID- 11430765 TI - An efficient method to successively introduce transgenes into a given genomic locus in the mouse. AB - BACKGROUND: Expression of transgenes in mice requires transcriptional regulatory elements that direct expression in a chosen cell type. Unfortunately, the availability of well-characterized promoters that direct bona-fide expression of transgenes in transgenic mice is limited. Here we described a method that allows highly efficient targeting of transgenes to a preselected locus in ES cells. RESULTS: A pgk-LoxP-Neo cassette was introduced into a desired genomic locus by homologous recombination in ES cells. The pgk promoter was then removed from the targeted ES cells by Cre recombinase thereby restoring the ES cells' sensitivity to G418. We demonstrated that transgenes could be efficiently introduced into this genomic locus by reconstituting a functional Neo gene. CONCLUSION: This approach is simple and extremely efficient in facilitating the introduction of single-copy transgenes into defined genomic loci. The availability of such an approach greatly enhances the ease of using endogenous regulatory elements to control transgene expression and, in turn, expands the repertoire of elements available for transgene expression. PMID- 11430766 TI - Intake and status in healthy Havana men, 2 years after the Cuban epidemic neuropathy. PMID- 11430767 TI - The cholesterolaemic effects of dietary fats in cholesteryl ester transfer protein transgenic mice. AB - In order to investigate the role of cholesteryl ester transfer protein (CETP) in the cholesterolaemic response to dietary fats, we analysed plasma lipid profiles of CETP-transgenic and control C57BL/6 mice fed standard chow (AIN-93G; AIN), a low-fat diet, and diets high in butter (saturated fatty acids; SFA), high-oleic acid safflower oil (monounsaturated fatty acids; MUFA), and safflower oil (polyunsaturated fatty acids; PUFA) for 5 weeks. Each group contained four or five mice. There were significant diet and dietxgenotype effects on plasma total cholesterol (TC; and respectively), liver TC ( and respectively), and esterified cholesterol (EC; and respectively); diet effects on plasma triacylglycerol liver free cholesterol and body weight a genotype effect on body-weight gain and a dietxgenotype effect on energy intake In transgenic mice the SFA diet caused significantly higher plasma TC than the PUFA diet In control mice MUFA and PUFA diets, but not the SFA diet, caused significantly higher plasma TC than the low fat and AIN diets Transgenic mice fed PUFA had lower plasma TC while transgenic mice fed MUFA had lower LDL+VLDL-cholesterol than controls in the same dietary groups. Transgenic mice fed MUFA and PUFA diets also had significantly higher liver TC and respectively) and EC and respectively) than controls fed the same diets. In the present study we showed that: (1) CETP transgenic mice had a cholesterolaemic response to dietary fats similar to that in human subjects; (2) CETP transgenic mice fed PUFA showed significantly lower plasma TC, while those fed MUFA had lower LDL+VLDL-cholesterol than controls; (3) hepatic accumulation of cholesterol, possibly resulting from the combination of the enhanced cholesteryl ester transfer to apolipoprotein B-containing lipoproteins and increased hepatic uptake of cholesterol, may contribute to the cholesterol lowering effect of MUFA and PUFA in CETP-transgenic mice; (4) CETP may play a role in appetite and/or energy regulation. PMID- 11430768 TI - Ontogenesis of hexokinase I and hexokinase IV (glucokinase) gene expressions in common carp (Cyprinus carpio) related to diet. AB - The expressions of hexokinase IV (glucokinase, GK) and hexokinase (HK)-I genes were analysed during early ontogenesis of common carp (Cyprinus carpio). Unlike HK-I gene, which was expressed during all the stages of the development, GK was only induced by the first feeding with high levels of dextrin as a source of carbohydrate. This study confirms the high capacity of common carp to use glucose even at the very early stages of development. PMID- 11430769 TI - Re-evaluation of the metabolism of oral doses of racemic carbon-6 isomers of formyltetrahydrofolate in human subjects. AB - The racemic mixture, [6RS]-5-formyltetrahydrofolate, is widely used clinically. In human subjects, orally-administered pure unnatural C-6 isomers, [6R]-5 formyltetrahydrofolate and [6S]-5,10-methenyltetrahydrofolate, were recently shown to be metabolized to the natural isomer, [6S]-5-methyltetrahydrofolate. We re-analysed the data from human studies published during the past four decades in which oral doses (< or =10 mg) of racemic mixtures of these folates were used. We re-evaluated the data to determine whether these racemic mixtures are only 50 % bioactive or, as we now predict, more than 50 % bioactive. Our analyses indicate that, in human subjects, oral doses of the racemic mixture of the two formyltetrahydrofolates are 20-84 % more bioactive than would be predicted. These data are consistent with the following pathway: chemical conversion of these folates to 10-formyltetrahydrofolate; oxidation of 10-formyltetrahydrofolate to 10-formyldihydrofolate; subsequent enzymic conversion of 10-formyldihydrofolate to dihydrofolate by 5-amino-4-imidazolecarboxamide ribotide transformylase; and finally the well-established metabolism of dihydrofolate to [6S]-5 methyltetrahydrofolate. An additional review of the literature supports the in vivo oxidation of 10-formyltetrahydrofolate occurring to a certain extent, as 10 formyl-folic acid is rapidly formed after the administration of folic acid (pteroylglutamic acid) or 5-formyltetrahydrofolate in human subjects. The dogma that an oral dose of the unnatural C-6 isomer of 5-formyltetrahydrofolate is not bioactive in human subjects does not withstand scrutiny, most probably due to the previously unrecognized in vivo oxidation of 10-formyltetrahydrofolate. This discovery unveils new folate metabolism in human subjects. PMID- 11430770 TI - Rapeseed glucosinolates and iodine in sows affect the milk iodine concentration and the iodine status of piglets. AB - I in the chain sow diet --> blood serum of sow --> sow milk --> piglet serum was investigated in two experiments with a total of eighty-one sows and their piglets. In experiments conducted during the last trimester of gravidity and the 28 d of lactation, diets with glucosinolates (1.9 mmol/kg diet via 100 g ground rapeseed/kg diet (Expt 1) and 2.1 and 4.2 mmol/kg diet via 75 and 150 g rapeseed press cake/kg diet (Expt 2)) were compared with control groups without rapeseed products. From 0 to 600 microg I/kg was added to sow diets during lactation. Diets without supplementary I decreased the I concentration particularly in milk and piglet serum. The presence of rapeseed and rapeseed press cake were indicated by a thiocyanate concentration increase, mainly in sow serum. The diets with glucosinolates decreased the milk and piglet serum I concentration. Spot urine and faeces samples from sows eating the rapeseed-press cake diets had increased I concentration. The sows' serum I and thyroxine did not respond to glucosinolates (Expt 1) or these diets caused an increase in concentration (Expt 2). Both these criteria seem unsuitable for the diagnosis of I status of adult animals. Glucosinolates and their degradation compounds may affect the thyroid and the mammary glands resulting in lower I milk transfer and higher renal and intestinal I excretion. PMID- 11430771 TI - Metabolic response to small and large 13C-labelled pasta meals following rest or exercise in man. AB - The metabolic response to a 150 or 400 g 13C-labelled pasta meal was studied for 8 h following rest or exercise at low or moderate workload (n 6). Following rest, the 400 g meal totally suppressed fat oxidation (v. 14.1 g following the 150 g meal) and a small amount of glucose was converted into fat (4.6 g), but fat oxidation remained high in subjects who had exercised following both the small (21.8 and 34.1 g) and large meal (14.1 and 32.3 g). Exogenous glucose oxidation was significantly higher in subjects who had remained at rest both following the small (67.6 g v. 60.4 and 51.3 g in subjects who exercised at low and moderate workloads) and large meal (152.2 v. 123.0 and 127.2 g). Endogenous glucose oxidation was similar in the three groups following the 150 g meal (42.3-58.0 g), but was significantly lower following the 400 g meal in subjects who had exercised at low workload (24.2 v. 72.2 g following rest; and was totally suppressed in those who had exercised at moderate workload. As a consequence, a larger positive glycogen balance was observed in subjects who exercised before the large meal (182.8-205.1 g v. 92.4 g following rest; Total fat oxidation calculated from 08.00 hours to 20.00 hours was similar in subjects who exercised at low and moderate workloads. These results indicate that: (1) de novo lipogenesis, which plays only a minor role for the disposal of an acute dietary carbohydrate load, is totally suppressed following exercise, even when a very large carbohydrate load is ingested; (2) the reduction in glycogen turnover as well as a preferential conversion of glucose into glycogen are responsible for the increase in glycogen stores following exercise; (3) for a similar energy expenditure, exercise at low workload for a longer period does not favour fat oxidation when the post-exercise period is taken into account. PMID- 11430772 TI - Iron loading and large doses of intravenous ascorbic acid promote lipid peroxidation in whole serum in guinea pigs. AB - Large doses of ascorbic acid may mobilise Fe from Fe-binding proteins in vivo which in turn could catalyse lipid peroxidation, a process associated with degenerative diseases. This hypothesis was tested in vitro in the serum of Fe loaded animals. Eighteen male guinea pigs weighing about 500 g on arrival were allocated to two groups of nine. Fe loading was induced in one group by two intraperitoneal injections of 200 mg iron dextran given on days 1 and 5. Blood (6 ml) was drawn from all animals on day 12 by cardiac puncture. Serum and LDL were separated. Serum was tested for loosely-bound Fe (bleomycin assay) and lipid peroxidation (thiobarbituric acid reactive substances (TBARS) assay) and LDL for susceptibility to in vitro oxidation (TBARS and conjugated diene assays). On day 12, another intraperitoneal injection of 200 mg iron dextran was given to the animals in the Fe-loaded group. On day 19, all animals were given 75 mg ascorbic acid by intraperitoneal injection. Blood (6 ml) was drawn 4 h later by cardiac puncture. Serum and LDL assays were repeated. Ascorbic acid increased loosely bound Fe and in vitro oxidation in the serum from animals of the Fe-loaded group but not in the serum from animals of the control group. Susceptibility of LDL to in vitro oxidation increased after the ascorbic acid injection in the control group but there was no further increase in the Fe-loaded group. These data suggest that large doses of ascorbic acid promote Fe mobilisation and in vitro oxidation in the serum of Fe-loaded animals. PMID- 11430773 TI - Response in hepatic removal of amino acids by the sheep to short-term infusions of varied amounts of an amino acid mixture into the mesenteric vein. AB - Under conditions of chronic supply the liver removes most amino acids (AA) in excess of net anabolic needs. Little information is available, however, on how acute alterations in AA supply (as might occur with once-daily feeding regimens) are controlled by the liver. Are these also extracted completely in a 'first pass' manner or are there limitations to hepatic uptake? Furthermore, is the rate of removal 'saturable' (by Michaelis-Menten kinetics) over the range of supply experienced under normal feeding conditions? These questions have been addressed in a study that involved acute (4.5 h) increases in AA supply. Four sheep were prepared with trans-hepatic vascular catheters and were offered a basal diet (equivalent to 1.6xenergy maintenance) throughout. On four occasions, at 7 d intervals, they were infused with various amounts of an AA mixture into the mesenteric vein over a 4.5 h period. The mixture contained fourteen AA in the proportions present in rumen microbial protein. The amounts infused were calculated to provide an additional one, two, three and four times that absorbed from the basal diet. Continuous blood collections were removed over 2 h intervals before (basal diet only) and at 0.5-2.5 and 2.5-4.5 h of AA infusion. Transfers of AA, from the digestive tract and to the liver, were calculated for both plasma and total blood. The recovery of the infused AA across the portal-drained viscera (PDV) was quantitative (100%) only for histidine and proline, the remaining AA were recovered at 56-83 %. These losses correlated with the arterial concentrations and were probably due to removal of AA from the systemic circulation by the tissues of the digestive tract. Despite the wide range of net PDV appearances (i.e. absorbed plus infused), the percentage of most AA removed by the liver remained constant, but the percentage varied with AA (from 34 for proline to 78 for tryptophan for blood transfers). Thus, even when supply was increased 5-fold over baseline there was no indication that the transport into the liver declined, indeed the absolute removals continued to increase. In contrast, the branched-chain AA (isoleucine, leucine and valine) did not exhibit constant percentage extractions. Their percentage extractions were always the lowest (16, 10 and 25 respectively) and tended to decline at the highest infusion rates, indicative of saturation in hepatic transport and/or metabolism. The arterial concentrations of all infused AA increased with rate of infusion, again indicative that the liver did not extract all the net AA available across the PDV. Absolute amounts removed were similar between plasma and blood, indicating that most of the hepatic transfers occurred from plasma. The fractional rates of transfer from total inflow to the liver (i.e. with re-circulated AA included) were 3- to 4-fold lower than rates based on the amounts absorbed plus infused. The highest percentage extraction for total blood inflows was for serine (27), but most were between 6 and 16, except for the branched-chain AA, which were all <1. Use of percentage extractions based on total inflows are probably more appropriate for development of mathematical models of liver metabolism, and the current data suggest that constant values may be applied. The needs of the liver for specific mechanisms involving phenylalanine and histidine (plasma protein synthesis), glycine (detoxification of xenobiotics) and alanine (gluconeogenesis) probably also require to be included in such models. PMID- 11430774 TI - Feeding dried purple laver (nori) to vitamin B12-deficient rats significantly improves vitamin B12 status. AB - To clarify the bioavailability of vitamin B12 in lyophylized purple laver (nori; Porphyra yezoensis), total vitamin B12 and vitamin B12 analogue contents in the laver were determined, and the effects of feeding the laver to vitamin B12 deficient rats were investigated. The amount of total vitamin B12 in the dried purple laver was estimated to be 54.5 and 58.6 (se 5.3 and 7.5 respectively) microg/100 g dry weight by Lactobacillus bioassay and chemiluminescent assay with hog intrinsic factor respectively. The purple laver contained five types of biologically active vitamin B12 compounds (cyano-, hydroxo-, sulfito-, adenosyl- and methylcobalamin), in which the vitamin B12 coezymes (adenosyl- and methylcobalamin) comprised about 60 % of the total vitamin B12. When 9-week-old vitamin B12-deficient rats, which excreted substantial amounts of methylmalonic acid (71.7(se 20.2) micromol/d) in urine, were fed the diet supplemented with dried purple laver (10 microg/kg diet) for 20 d, urinary methylmalonic acid excretion (as an index of vitamin B12 deficiency) became undetectable and hepatic vitamin B12 (especially adenosylcobalamin) levels were significantly increased. These results indicate that vitamin B12 in dried purple laver is bioavailable to rats. PMID- 11430775 TI - Gastrointestinal handling and metabolic disposal of 13C-labelled tripalmitin during rehabilitation from childhood malnutrition. AB - We investigated the gastrointestinal handling and post-absorptive metabolic handling of [1,1,1-13C]tripalmitin and [1-13C]glycocholate during recovery from severe childhood malnutrition. Eight children were studied on three occasions: at admission (phase 1), during rapid catch-up growth (phase 2) and when weight-for height had reached 90 % of the reference (phase 3). Breath samples were obtained over a 24 h period and stools were collected over 3 d following the administration of each tracer. At admission, the lipid content of stool expressed as a percentage of ingested lipid was 6 (range 0.7-28.9) but less variation was shown between children at phase 2 (3.3 (range 0.9-4.1)) and phase 3 (1.4 (range 0.4-2.5)). The excretion of 13C in stool varied markedly between children at admission (11.1 (sd 5.4) % administered dose) and during rehabilitation (phase 2, 15.4 (sd 16.5) % administered dose; phase 3, 6.2 (sd 10.2) % administered dose). About 5 % of the absorbed label was recovered on breath at each stage (% absorbed dose; phase 1, 5.1 (sd 6.0); phase 2, 5.2 (sd 3.1); phase 3, 6.4 (sd 6.6)). None of the children exhibited significant bile salt malabsorption as a consequence of small intestinal overgrowth. Of the 13C measured in stool, more label was recovered in fatty acids than triacylglycerols during each of the three phases and this was interpreted to reflect a failure to absorb the products of digestion. The results show that not all the children had problems associated with the digestion and absorption of 13C-labelled tripalmitin in severe malnutrition and during recovery, which was not reflected in gross lipid balance across the gastrointestinal tract. Absorbed lipid was more likely to be deposited as adipose tissue than to satisfy the immediate needs for energy. PMID- 11430776 TI - Evidence for the existence of adaptive thermogenesis during weight loss. AB - The present study was performed to further investigate the adaptive component of thermogenesis that appears during prolonged energy restriction. Fifteen obese men and twenty obese women underwent a 15-week weight-loss programme. During this programme, body weight and composition as well as resting energy expenditure (REE) were measured at baseline, after 2 and 8 weeks of energy restriction (-2929 kJ/d) and drug therapy (or placebo), and finally 2-4 weeks after the end of the 15-week drug therapy and energy restriction intervention, when subjects were weight stable. Regression equations were established in a control population of the same age. These equations were then used to predict REE in obese men and women at baseline, after 2 and 8 weeks, as well as after the completion of the programme. In both men and women body weight and fat mass were significantly reduced in all cases) while fat-free mass remained unchanged throughout the programme. At baseline, REE predicted from the regression equation was not significantly different from the measured REE in men, while in women the measured REE was 13 % greater than predicted. After 2 weeks of energy restriction, measured REE had fallen by 469 and 635 kJ/d more than predicted and this difference reached 963 and 614 kJ/d by week 8 of treatment in men and women respectively. Once body-weight stability was recovered at the end of the programme, changes in REE remained below predicted changes in men (-622 kJ/d). However, in women changes in predicted and measured REE were no longer different at this time, even if the women were maintaining a reduced body weight. In summary, the present results confirm the existence of adaptive thermogenesis and give objective measurements of this component during weight loss in obese men and women, while they also emphasize that in women this component seems to be essentially explained by the energy restriction. PMID- 11430777 TI - Are the proposed limits of energy intake:basal metabolic rate and dietary nitrogen:urinary nitrogen ratios suitable for validation of food intake? AB - The validity of 7 d weighed records of diet obtained for pre-menopausal Mexican women was assessed by two independent methods: the energy intake:BMR (EI:BMR) and the dietary N:urinary N (DN:UN). For the latter, complete urine collections are required and completeness was assessed from measurements of para-aminobenzoic acid (PABA) excretion. There were forty-six adult female subjects in the study, thirty-four were from Mexico City and twelve were from a rural population in the Central Highlands, Mexico. However, data were rejected from five urban women for whom the PABA excretion data suggested incomplete urine collection on four or more days. BMR was measured with Oxylog portable O2 consumption meters, and physical activity level was assessed from a self-completed activity diary. An approximate relationship between the EI:BMR ratio and the DN:UN ratio suggested that the rejection limits on the EI:BMR ratio recommended by are wider than the limits on the DN:UN ratio recommended by. Using the recommended cut-off points for EI:BMR but wider limits for DN:UN, twenty-one and twenty-five women respectively had acceptable intake records by the two methods, and sixteen of them by both methods. In conclusion the modification of the DN:UN limits to 0.92 and 1.70 to set acceptable intake values makes the use of measurements of N and energy balance comparable. Urine values with PABA recoveries greater than should be rejected, as should UN values validated by less than 3 d. PMID- 11430778 TI - Nutritional status in elderly female hip fracture patients: comparison with an age-matched home living group attending day centres. AB - Fractured neck of femur occurs mostly in the older female population and is generally caused by falls. Malnutrition has been postulated as a factor that increases the tendency to suffer falls. Nutritional status of older female hospital patients admitted for emergency surgery for fractured neck of femur recruited (n 75), was compared with an age-matched independent-living group of females attending one of three local day centres (n 50). Dietary assessment was undertaken using three consecutive 24 h dietary recalls and, in the hip fracture group, completed menu cards were used as memory prompts. Data concerning key lifestyle characteristics were obtained using a face-to-face administered questionnaire. Blood samples were taken to determine levels of plasma albumin, transferrin, C-reactive protein (CRP), cholesterol, vitamin C, Se, Zn and total antioxidant status. Haemolysate samples were analysed for Se-dependent glutathione peroxidase activity. There were no significant differences in age between the two groups, but the hip fracture patients had lower mean values for body weight (59.6 v. 67.5 kg; mindex (weight/demispan) (83.1 v. 94.4 kg/m; calculated BMI (24.1 v. 27.5 kg/m2 mid-upper arm circumference; 27.1 v. 31.3 cm, and triceps skinfold thickness; 17.0 v. 18.9 mm, than the home-living group. The hip fracture patients had lower intakes of energy (4.3 v. 5.4 MJ, fat carbohydrate protein thiamine vitamin B6 calcium K Mg P Fe Se and NSP Mean intakes of both groups were below the estimated average requirement for energy and below the reference nutrient intakes for folate, Ca, vitamin D, Mg, K, Se and Zn. In a high percentage of the hip fracture group the dietary intake of particular nutrients fell below the lower reference nutrient intake for Se (73 %), Mg (54 %) and Fe (19 %). As expected, the fracture patients had reduced plasma albumin and increased CRP values. They had higher plasma vitamin C levels and lower cholesterol levels than the day centre attendees. There were no significant differences in plasma levels of Se, Zn, transferrin or haemolysate glutathione peroxidase activity between the two groups. However, there was evidence of under-nutrition in both groups as key anthropometric values were low, plasma nutrient and metabolite levels were below the standard reference ranges and many individuals had low dietary intakes for specified nutrients. PMID- 11430779 TI - Vitamin B intake and status in healthy Havanan men, 2 years after the Cuban neuropathy epidemic. AB - A prospective epidemiological study was carried out over 1 year to evaluate vitamin B complex dietary intake and status in Cuba, 2 years after the Cuban neuropathy epidemic of 1993. Of the 199 healthy middle-aged men selected, 141 completed the study. Volunteers were followed up every 3 months for 1 year. Dietary intake and status of thiamin, riboflavin, vitamin B6, folate and vitamin B12 were assessed each time. The dietary intake of vitamin B complex was low, particularly in June and July (folate), and October (thiamin). A deficient status was observed for vitamin B complex, except for vitamin B6. Vitamin B complex intake and status varied over the year. However, dietary intake and status were poorly related. The results prove that healthy Cuban men represent a vulnerable population in terms of vitamin B complex status and stress the necessity to both promote preventive multivitamin supplementation and produce local food rich in vitamin B complex. PMID- 11430780 TI - Low serum lycopene concentration is associated with an excess incidence of acute coronary events and stroke: the Kuopio Ischaemic Heart Disease Risk Factor Study. AB - A number of epidemiological studies have shown an association between beta carotene and the risk of cardiovascular diseases, whereas only a few studies are available concerning the association of lycopene with the risk of coronary events, and no studies have been undertaken concerning lycopene and stroke. Thus, we tested the hypothesis that low serum levels of lycopene are associated with increased risk of acute coronary events and stroke in middle-aged men previously free of CHD and stroke. The subjects were 725 men aged 46-64 years examined in 1991-3 in the Kuopio Ischaemic Heart Disease Risk Factor Study. Forty-one men had either a fatal or a non-fatal acute coronary event or a stroke by December 1997. In a Cox' proportional hazard's model adjusting for examination years, age, systolic blood pressure and three nutritional factors (serum folate, beta carotene and plasma vitamin C), men in the lowest quarter of serum lycopene levels (< or =0.07 micromol/l) had a 3.3-fold (95 % CI 1.7, 6.4, risk of acute coronary events or stroke compared with the others. Our study suggests that a low serum level of lycopene is associated with an increased risk of atherosclerotic vascular events in middle-aged men previously free of CHD and stroke. PMID- 11430781 TI - Linear and knemometric growth in the early phase of rehabilitation from severe malnutrition. AB - The relationship between ponderal, linear and lower leg growth in children recovering from severe malnutrition remains unclear. We report on the early growth of 141 severely malnourished Bangladeshi children aged 6 to 36 months of age who were followed for 90 d. Mean (sd) weight for height (WHZ) and height for age (HAZ) catch-up growth z scores over the 90 d were 1.6 (0.85) and 0.47 (0.325) respectively. Mean (sd) lower leg length growth was 10.35 (4.5) mm. Change in HAZ was significantly associated with initial WHZ, but linear growth occurred in the presence of severe wasting and no threshold WHZ score was identified. Lower leg length gain correlated throughout with ponderal indices but with change in HAZ score only after day 45. Only initial WHZ score and maternal height predicted for linear growth and only accounted for 20 % of total variance. We conclude that linear growth occurs early in severely malnourished children but that knemometry behaves as a ponderal index acutely. PMID- 11430783 TI - Precision measurement of disc height, vertebral height and sagittal plane displacement from lateral radiographic views of the lumbar spine. AB - OBJECTIVE.: To compile a database of disc height, vertebral height and sagittal plane displacement from lateral radiographic views of the lumbar spine, valid for male and female subjects in the age range 16-57 years. The protocols used to measure these parameters compensate for distortion in central projection, off centre position, axial rotation and lateral tilt of the spine as well as for variation in radiographic magnification and stature. STUDY DESIGN.: The study comprised designing and testing of measurement protocols, together with subsequent data collection from archive radiographs. BACKGROUND.: Attempts to quantify primary mechanical damage to lumbar vertebrae and discs have been limited due to imprecision when measuring disc height, vertebral height and sagittal plane displacement. Age-related, normative values for these parameters were not previously available. Consequently, important issues like the effectiveness of past and present guidelines for safe manual handling with respect to prevention of overload injuries could not be resolved and judgement on pathological alterations in the morphology of the individual lumbar spine could only be performed in a qualitative, subjective manner. METHODS.: Based on the analysis of vertebral contours in the lateral radiographic image of the lumbar spine, new protocols for measuring disc height, vertebral height and sagittal plane displacement were developed. The measured data are virtually independent of distortion, axial rotation and lateral tilt. Furthermore, description of height and displacement using dimensionless parameters guarantees independence of radiographic magnification and stature. Subjective influence in the measurement procedure was minimized by automatic computation of contour-landmarks and derived parameters. Measurement errors were assessed from sets of radiographs of spine specimens and serial flexion-extension radiographs; interobserver and intraobserver errors were assessed from repeated measurements of lateral views. For compilation of a database, measurements were performed of a set of 892 lateral views of the lumbar spine of male and female subjects between 16 and 57 years of age. Data from pathologically deformed vertebrae or discs, or from motion segments exhibiting spondylolisthesis or retrolisthesis were excluded by normal radiological inspection; data from spines showing normal, age-related degenerative changes were included. RESULTS.: The new protocols allow height and displacement of lumbar vertebrae as well as height of lumbar discs to be measured for all motion segments on a lateral view, rather than just those vertebrae or discs close to the central beam, to facilitate recognition of localized abnormalities. Since the results are independent of exposure geometry, retrospective investigations are feasible. The relative measurement error in vertebral height amounts to 2.2%; for a vertebra of 30 mm height this corresponds to an error of approximately 0.7 mm. The error in sagittal plane displacement amounts to 0.015 (measured in units of mean vertebral depth); for a vertebra of 35 mm depth this corresponds to an error of 0.5 mm. The relative error in disc height amounts to 4.15%; for a disc of 10 mm height this corresponds to an error of approximately 0.5 mm. For both genders, the database contains age-dependent, normative values of disc height, vertebral height and sagittal plane displacement. In addition, the database describes intersegmental shape correlation, i.e. the relation between height of neighbouring discs and between height as well as displacement of neighbouring vertebrae. On average, height of lumbar vertebrae is larger in females than in males; height of lumbar discs is larger in males than in females and shows a minute dependence on age in males; in both genders, sagittal plane displacement increases, but only by a small amount, with age. CONCLUSIONS.: The new measurement protocols for disc height, vertebral height and sagittal plane displacement, together with the database of normative age-related values, permit quantitative assessment of the prevalence of pathological morphological changes in the human lumbar spine. The new method and the database will serve to explore the effect of potentially detrimental influences such as high spinal loading and to provide quantitative documentation of existing injury to vertebrae and discs in individual cases. PMID- 11430784 TI - Stiffness between different directions of transpedicular screws and vertebra. AB - OBJECTIVE: This study was undertaken to evaluate the variation in bone density within the vertebral body and to determine the biomechanical stiffness of the screw-bone interface for different superior-inferior transpedicular screw orientations in the vertebral body. DESIGN: The stiffness of three directions of screw placement (upper, middle, and lower) were measured in two modes of loading (flexion and torsion). All screws were inserted to 80% of the distance along the path of the screw from the point of insertion to the anterior vertebral cortex. BACKGROUND: The placement of transpedicular screws within the vertebral body is a fundamental determinant the stiffness and strength of the bone-screw interface and consequently the stability of an implant construct. The bone stiffness within the pedicle and vertebra must be sufficient to resist spine forces and moments. METHODS: The stiffness of pedicle-screw fixation was tested for three different screw orientations based on the position of the screw tip in the vertebral body (upper, middle, and lower). Fixation rigidity was measured for two modes of loading (flexion and torsion). In all, eighteen individual vertebrae (L(2)-L(4)) from 6 cadaveric spine segments were examined. All the screws were inserted to 80% of the depth of the posterior to anterior vertebral cortex dimension. Quantitative bone density measurements were obtained from vertebral mid-sagittal cross sections of the additional vertebral bodies using an image analysis system to determine the distribution of bone density in the mid-sagittal plane. RESULTS: Based on area fraction measurements of cancellous bone in seventeen L1 mid sagittal regions, highest densities were observed adjacent to the endplates (area fractions > 0.29). Regions of lowest density were found in the central portions of the vertebral bodies, above the mid-line of the body (area fractions < 0.20). In flexion, the stiffness of all three groups of screws were similar, ranging from 4.0-4.2 Nm deg(-1). In torsion the upper and middle directions were significantly more rigid than lower position (p < 0.04). CONCLUSION: Pedicle screw insertion aimed toward the superior-anterior aspect of the vertebral body, adjacent to the superior endplate provides the best overall rigidity considering both directions of loading. Screws placed in the mid-portion of the vertebral body may provide good lateral torsion rigidity, but would not be optimal for flexion stability if bone density is reduced in that region. PMID- 11430785 TI - Myo-electric behavior of the trunk muscles during static load holding in healthy subjects and low back pain patients. AB - OBJECTIVE: The purposes of this study were to examine coactivational activities of four trunk muscles in various static holding tasks, and to examine the differences of muscle activation patterns between normal subjects and low back pain (LBP) patients. DESIGN: We used an one-way analysis of variance (anova) with repeated measures to analyze the effect of the knee angles, trunk flexion angles, and loading conditions on the amount of EMG activity. BACKGROUND: Epidemiologic studies attribute a considerable proportion of the incidence of LBP to lifting related injuries. The relationship between LBP and trunk muscles integrated electromyography (IEMG) is controversial. METHODS: Forty volunteer subjects with no experience of LBP and 47 LBP patients recruited from Chung-Gung Memory Hospital participated in this experiment. Surface IEMG signals of four right and left trunk muscles (rectus abdominis, external oblique, erector spinae, and latissimus dorsi) were obtained in static holding tasks. RESULTS: In knee flexion, LBP patients had significant IEMG pattern change than normal subjects. In trunk flexion and loading conditions, normal subjects had significant IEMG pattern change than LBP patients. CONCLUSIONS: Trunk muscle groups have different muscle coactivation patterns in functional posture of load holding. Using surface EMG can discriminate LBP patients from normal subjects. PMID- 11430786 TI - The dynamic response of L(2)/L(3) motion segment in cyclic axial compressive loading. AB - OBJECTIVE: The dynamic response and load sharing amongst passive elements of an L2-L3 motion segment during axial compressive cyclic loading was investigated. DESIGN: A validated viscoelastic nonlinear finite element model of L2-L3 was used for a detailed stress/strain analysis during axial cyclic loading. BACKGROUND: The repetitive loading of the spine has been implicated as a risk factor in developing low back disorders. However, the quantitative description of injury mechanisms and the internal load sharing have been lacking. METHODS: The applied cyclic axial compressive loading was controlled, peak to peak, from 600 to 1000 N at 0.5 Hz for 15 cycles. The stress/strain and strain energy density of various elements were quantified and the effects of cyclic loading on these parameters were investigated. RESULTS: The axial stiffness of the motion segment decreased, while intradiscal pressure (IDP) and the strain in anulus fibers of the outermost lamella increased. The axial stresses of outer lamellae in the anulus matrix reduced, in contrast to the increased strain at the endplate. CONCLUSIONS: The load sharing amongst the passive elements of the motion segment changed. The response of the motion segment to the same external axial load depends on the history of loading. The anulus fibers in the innermost layer were slack due to compression, hence not at risk of failure. The loss of disc height and increased disc bulge led to higher strain in anulus fibers of outermost layer. In future, more complex loading conditions with a longer duration should be considered. PMID- 11430787 TI - Effects of varying backpack loads on peak forces in the lumbosacral spine during walking. AB - OBJECTIVE: To compare the differences in lumbosacral spine forces under varying backpack loads. DESIGN: A biomechanical model was used to determine the changes in peak forces in the L5/S1 joint with increasing backpack loads during level walking. BACKGROUND: Most studies involving varying external backpack loads have been concerned mainly with kinematic and physiological measurements. To the author's knowledge, there has been no investigation of the change in peak forces in the lumbosacral joint during the carriage of such loads. METHOD: Data acquisition was carried out using a 5-camera Vicon motion analysis system and two Kistler force plates. Ten male subjects with similar weights, height and age were recruited for this study. Three different backpack loading conditions were studied, that is walking with no load, with 15% BW and with 30% BW. RESULTS: It was observed that all the ten subjects while walking with heavier backpack load adopted a compensatory trunk flexion posture. However, kinematic gait parameters such as walking speed and stride length remained unchanged with the increasing loads. Walking with backpack load of 15%BW and 30%BW resulted in corresponding increase in lumbosacral force of 26.7% and 64% respectively when compared to walking without backpack load. CONCLUSION: In carrying a given packload during walking, it will give rise to a disproportionate force increase acting on the L5/S1 joint. PMID- 11430788 TI - Rate-related fatigue injury of vertebral disc under axial cyclic loading in a porcine body-disc-body unit. AB - OBJECTIVE: Cyclic loading tests were performed on fresh mature porcine lumbar spines to investigate the mechanical responses at different loading rates, morphological changes in the disc and to produce the clinically relevant injuries of porcine intervertebral disc. DESIGN: Two-segment units of fresh porcine spine with all the posterior elements removed were used for a disc stress analysis and morphological observation during axial cyclic loading. BACKGROUND: The repetitive loading of the spine has been implicated as a risk factor in developing low back disorders. However, few studies have discussed morphologic changes in vertebral disc due to fatigue stress and no study has investigated the relationship between disc strength and bone mineral density. METHODS: 21 body-disc-body specimens divided into three groups were subjected to cyclic loading at test speeds of 0.5, 5 and 20 mm min(-1). The correlation between residual stress of specimen and bone mineral density was investigated and the disc morphological changes were observed through the middle cross section of the intervertebral disc using a zoom stereo microscope. RESULTS: There was no visible crack observed following testing in the 0.5 mm min(-1) test group. However, one disc in the 5 mm min(-1) and two discs in the 20 mm min(-1) test group exhibited disc herniation. The residual stress was found to be positively and linearly correlated with bone mineral density and decreased as the loading rates increased. CONCLUSION: Faster loading rate generates greater stress decay, and disc herniation is more likely to occur under higher loading rate conditions. The most common site of disc herniation is the posterolateral area which is in agreement with that of the human spine. PMID- 11430789 TI - Biomechanical evaluation of a new anterior spinal implant. AB - OBJECTIVE: To biomechanically evaluate the construct of a new anterior spinal implant for osteoporotic patients. DESIGN: Mechanical tests and finite element analysis were designed to evaluate the spinal implant. BACKGROUND: Many osteoporotic thoracolumbar spine fractures often need surgical intervention to relieve pains, stabilize progressive kyphosis or improve neurologic impairment. Progressive kyphosis, instrumentation failure and pseudoarthrosis are often seen post-operatively. These results may be due to insufficient bonding between the screw and the vertebral body. As the end-plate is the most rigid part of the vertebral body, the spinal implant was designed so that the end-block would fix onto the end-plates of the vertebral bodies. METHODS: Two biomechanical evaluations of the new anterior spinal implant were conducted to evaluate the construct stiffness and the bonding strength between the spinal implant and the vertebral body. In the evaluation of the construct stiffness, the biomechanical tests between the new spinal implant and the Kaneda device were performed on six fresh intact porcine spines at the thoracolumbar region. In the prediction of the loading transfer between the spinal implant and the vertebral body, a three dimensional finite element model (FEM) was built to simulate the osteoporotic vertebral body and the new anterior spinal implant. RESULTS: The results of the mechanical tests showed that the compressive stiffnesses of the Kaneda device and the new implant were 357 +/- 37 N mm(-1) and 297 +/- 98 N mm(-1). Their flexional stiffnesses were 0.339 +/- 0.126 N-m mm(-1) and 0.364 +/- 0.107 N-m mm(-1), respectively. Their torsional stiffnesses were 6.37 +/- 0.28 N-m deg(-1) and 5.30 +/- 0.71 N-m deg(-1), respectively. There were no significant differences (p > 0.01). The results of FEM showed that the new implant had high stress concentration on the junction of the screw and plate, screw and end-blocks, and between the end-plate and the end-blocks. CONCLUSIONS: The stabilization effect of the new implant was similar to that of the Kaneda device. With less rigid mechanical properties, the new implant can be more beneficial to the remodeling process of the spinal structure after instrumentation. The new anterior spinal implant showed a high potential for application to osteoporotic patients. PMID- 11430790 TI - Isokinetic evaluation of the ankle before and after surgical treatment of the lumbar disc herniation. AB - OBJECTIVE: To quantitatively evaluate the ankle strength before and after surgical treatment for herniated lumbar intervertebral disc. DESIGN: Isokinetic test of ankle joint and statistical analysis were designed to evaluate the results before and after surgery. BACKGROUND: Assessments of the results before and after treatment of lumbar herniation of intervertebral discs (HIVD) in previous papers were all based on patients' symptoms, activity, working condition, the angle of straight leg raising test, and requirement of pain medication. We designed a prospective study of isokinetic measurement of the ankle joints in those patients with HIVD to try to find a quantitative method of evaluation. METHODS: The criteria of selecting patients were single level disc herniation of lumber spine, without associated major neuromuscular or musculoskeletal disorder, normal mental status and cooperative, clinically evident radiculopathy, failure of conservative treatment, and age under 45. The isokinetic strengths of the ankle joints, of both involved and uninvolved limbs, were evaluated with plantarflexion or dorsiflexion, in the velocities of 30, 60, 120, and 180 degrees s(-1), according to their affected levels. Standard laminotomy with removal of intervertebral disc was undertaken for all patients under general anesthesia. The isokinetic strength of the ankle joints were evaluated with Kin-Com dynamometer before surgery, and post-operatively at 1 week, 2 weeks, 4 weeks, 2 months, and 3 months. All data were collected and analyzed with linear mixed model for analysis of variance with repeated measures. RESULTS: From January, 1996, 5 patients entered this study, 2 patients were L4-L5 herniation, and 3 for L5-S1. The estimated mean strength ratio and standard error was 55.04 +/- 2.21% preoperatively, 46.42 +/- 2.22% postoperatively at 1 week, 64.35 +/- 2.23% postoperatively at 2 weeks, 73.93 +/- 2.23% postoperatively at 4 weeks, 100.40 +/- 2.22% postoperatively at 2 months, and 104.55 +/- 2.21% postoperatively at 3 months. CONCLUSIONS: In this preliminary prospective study, there seems no correlation between duration of the symptoms and isokinetic strength of the ankle joint. There is, however, marked improvement of isokinetic strength of ankle joint postoperatively at 2 months, which was statistically significant. PMID- 11430791 TI - The postural stability control and gait pattern of idiopathic scoliosis adolescents. AB - OBJECTIVE: The static postural equilibrium and gait patterns between idiopathic scoliotic (IS) patients and normal subjects were studied to verify the best method to identify the functional disability in IS patients. DESIGN: The static stability in six postures and gait patterns among normal subjects and IS patients were compared. BACKGROUND: Postural stability control and gait analysis are non invasive methods to identify many diseases. However, the dysfunction of IS patients in postural stability control and gait pattern is not clear. The results of this research may lead to further understanding of the etiology of idiopathic scoliosis in the postural equilibrium influencing aspects. METHODS: Thirty IS patients and fifteen normal subjects were recruited for postural stability control test and gait analysis using the force plate and 3-D motion analysis system. RESULTS: The IS patients generally produced higher sway area, lateral sway, sagittal sway, and sway radius than normal subjects. The cadence is smaller in the IS patients, but the stance phase and stride phase are similar to normal subjects. CONCLUSIONS: The IS patients are poor in postural stability control but their gait pattern is similar to that of normal subjects. Standing with trunk at full flexion is the most effective position to identify the postural stability control of IS patient. PMID- 11430792 TI - A three-dimensional mathematical model for predicting spinal joint force distribution during manual liftings. AB - OBJECTIVE: A three-dimensional dynamic mathematical model was developed to discover what loads are imposed on the lumbar structures by performance of asymmetric manual liftings. DESIGN: An external model was used to estimate the intersegmental resultant forces and moments at the L(5)/S(1) joint in this dynamic biomechanical model. Using an optimization algorithm, an internal model then distributed the intersegmental resultants to forces of muscle, disc, facet joints, and ligaments. BACKGROUND: To study the relation between large loads and low-back disorders, many biomechanical models have been developed. Most of the models were two-dimensional models discussed with symmetric activities. Some three-dimensional biomechanical models were static models or only included limited elements of the disc and muscles in the model. METHODS: A healthy young male subject was asked to perform asymmetric lift with bent knees. Dynamic data of body motion and ground reaction forces were monitored, and the EMG of six muscles were recorded simultaneously. A Newtonian equation was used to calculate the joint intersegmental resultant forces and moments. In the internal model, three components of the disc force, eight muscle forces, two ligament forces and two facet joint forces were computed. RESULTS: The correlation between the reaction moments from the upper and lower models of the external part were generally above 0.94, and the root mean square differences were below 19 Nm. In this internal model, the maximal disc compression was close to the data showed on the literature, and the estimation of muscle forces corresponded to the EMG activities. CONCLUSIONS: A three-dimensional biomechanical model has been developed and evaluated to estimate the spinal joint force distribution during asymmetric manual lifting activities. PMID- 11430793 TI - Quantification of overload injuries to thoracolumbar vertebrae and discs in persons exposed to heavy physical exertions or vibration at the workplace Part II Occurrence and magnitude of overload injury in exposed cohorts. PMID- 11430794 TI - Metal chelator decreases Alzheimer beta-amyloid plaques. AB - Transgenic mice developing beta-amyloid (Abeta) plaques are advancing experimental treatment strategies for Alzheimer's disease. The metal chelator, clioquinol, is reported by Cherny et al. (2001) to reduce Abeta plaques, presumably by chelation of Abeta-associated zinc and copper. This and other recent Abeta-modulating treatment approaches are discussed. PMID- 11430795 TI - Do Schwann cells stop, DR(o)P2, and roll? AB - Dystrophin-dystroglycan complexes in Schwann cells may play a role in both signaling and structural interactions between the extracellular matrix and the cytoskeleton. Sherman et al. (2001) show that a new complex containing dystrophin related protein 2 (DRP2) and periaxin plays a role in Schwann cell-basal lamina interactions and PNS myelination. PMID- 11430796 TI - Knock out of direction selectivity in the retina. AB - Retinal ganglion cells show direction selectivity in their responses to moving stimuli. The circuitry necessary to generate directional selectivity in these cells has been long debated. Yoshida et al. (2001) use immunotoxin-mediated cell ablation to demonstrate that the starburst amacrine cell is at the core of this computation. PMID- 11430797 TI - Reverse spikeology: predicting single spikes. AB - Neural models that simulate single spike trains can help us understand the basic principles of neural coding in vision. Keat et al. (2001) develop a hybrid model that combines spatiotemporal filtering with nonlinear spike generation. The model does a good job of predicting the responses of single retinal ganglion cells and thalamic relay neurons. PMID- 11430798 TI - Methylated cytosine and the brain: a new base for neuroscience. AB - Methylation of genomic CpG residues is crucial for proper neuronal function. Rett syndrome, a common form of mental retardation, is associated with mutations in the gene encoding MeCP2, a methyl CpG binding protein linked to transcriptional repression. Gene knockouts of mouse Mecp2 have reproduced key aspects of the disease. A CNS-restricted knockout of Dnmt1, encoding the enzyme that maintains CpG methylation patterns, results in loss of mutant neurons and glia. PMID- 11430799 TI - Mushroom bodies, Ca(2+) oscillations, and the memory gene amnesiac. AB - The memory of odors in Drosophila is mediated by mushroom body neurons. Memory is formed, in part, by a modulation of the physiology of these neurons brought about by neuropeptides that are encoded by the amnesiac gene and released from peptidergic neurons that innervate mushroom body neurons. Slow and spontaneous oscillations of calcium levels are elevated in the mushroom body neurons of amnesiac mutants and may contribute to memory consolidation. PMID- 11430800 TI - Cross-modal plasticity underpins language recovery after cochlear implantation. AB - Postlingually deaf subjects learn the meaning of sounds after cochlear implantation by forming new associations between sounds and their sources. Implants generate coarse frequency responses, preventing place-coding fine enough to discriminate sounds with similar temporal characteristics, e.g., buck/duck. This limitation imposes a dependency on visual cues, e.g., lipreading. We hypothesized that cross-modal facilitation results from engagement of the visual cortex by purely auditory tasks. In four functional neuroimaging experiments, we show recruitment of early visual cortex (V1/V2) when cochlear implant users listen to sounds with eyes closed. Activity in visual cortex evolved in a stimulus-specific manner as a function of time from implantation reflecting experience-dependent adaptations in the postimplant phase. PMID- 11430801 TI - Treatment with a copper-zinc chelator markedly and rapidly inhibits beta-amyloid accumulation in Alzheimer's disease transgenic mice. AB - Inhibition of neocortical beta-amyloid (Abeta) accumulation may be essential in an effective therapeutic intervention for Alzheimer's disease (AD). Cu and Zn are enriched in Abeta deposits in AD, which are solubilized by Cu/Zn-selective chelators in vitro. Here we report a 49% decrease in brain Abeta deposition (-375 microg/g wet weight, p = 0.0001) in a blinded study of APP2576 transgenic mice treated orally for 9 weeks with clioquinol, an antibiotic and bioavailable Cu/Zn chelator. This was accompanied by a modest increase in soluble Abeta (1.45% of total cerebral Abeta); APP, synaptophysin, and GFAP levels were unaffected. General health and body weight parameters were significantly more stable in the treated animals. These results support targeting the interactions of Cu and Zn with Abeta as a novel therapy for the prevention and treatment of AD. PMID- 11430802 TI - Specific disruption of a schwann cell dystrophin-related protein complex in a demyelinating neuropathy. AB - Dystroglycan-dystrophin complexes are believed to have structural and signaling functions by linking extracellular matrix proteins to the cytoskeleton and cortical signaling molecules. Here we characterize a dystroglycan-dystrophin related protein 2 (DRP2) complex at the surface of myelin-forming Schwann cells. The complex is clustered by the interaction of DRP2 with L-periaxin, a homodimeric PDZ domain-containing protein. In the absence of L-periaxin, DRP2 is mislocalized and depleted, although other dystrophin family proteins are unaffected. Disruption of the DRP2-dystroglycan complex is followed by hypermyelination and destabilization of the Schwann cell-axon unit in Prx(-/-) mice. Hence, the DRP2-dystroglycan complex likely has a distinct function in the terminal stages of PNS myelinogenesis, possibly in the regulation of myelin thickness. PMID- 11430803 TI - Conformational changes in S6 coupled to the opening of cyclic nucleotide-gated channels. AB - In cyclic nucleotide-gated channels (CNG), direct binding of cyclic nucleotides in the carboxy-terminal region is allosterically coupled to opening of the pore. A CNG1 channel pore was probed using site-directed cysteine substitution to elucidate conformational changes associated with channel opening. The effects of cysteine modification on permeation suggest a structural homology between CNG and KcsA pores. We found that intersubunit disulfide bonds form spontaneously between S399C residues in the helix bundle when channels are in the closed but not in the open state. While MTSET modification of pore-lining residues was state dependent, Ag(+) modification of V391C, in the inner vestibule, occurred at the same diffusion-limited rate in both open and closed states. Our results suggest that the helix bundle undergoes a conformational change associated with gating but is not the activation gate for CNG channels. PMID- 11430804 TI - Phosphorylation of period is influenced by cycling physical associations of double-time, period, and timeless in the Drosophila clock. AB - The clock gene double-time (dbt) encodes an ortholog of casein kinase Iepsilon that promotes phosphorylation and turnover of the PERIOD protein. Whereas the period (per), timeless (tim), and dClock (dClk) genes of Drosophila each contribute cycling mRNA and protein to a circadian clock, dbt RNA and DBT protein are constitutively expressed. Robust circadian changes in DBT subcellular localization are nevertheless observed in clock-containing cells of the fly head. These localization rhythms accompany formation of protein complexes that include PER, TIM, and DBT, and reflect periodic redistribution between the nucleus and the cytoplasm. Nuclear phosphorylation of PER is strongly enhanced when TIM is removed from PER/TIM/DBT complexes. The varying associations of PER, DBT and TIM appear to determine the onset and duration of nuclear PER function within the Drosophila clock. PMID- 11430805 TI - Distinct subpopulations of sensory afferents require F11 or axonin-1 for growth to their target layers within the spinal cord of the chick. AB - Dorsal root ganglion neurons project axons to specific target layers in the gray matter of the spinal cord, according to their sensory modality. Using an in vivo approach, we demonstrate an involvement of the two immunoglobulin superfamily cell adhesion molecules axonin-1/TAG-1 and F11/F3/contactin in subpopulation specific sensory axon guidance. Proprioceptive neurons, which establish connections with motoneurons in the ventral horn, depend on F11 interactions. Nociceptive fibers, which target to layers in the dorsal horn, require axonin-1 for pathfinding. In vitro NgCAM and NrCAM were shown to bind to both axonin-1 and F11. However, despite this fact and despite their ubiquitous expression in the spinal cord, NgCAM and NrCAM are selective binding partners for axonin-1 and F11 in sensory axon guidance. Whereas nociceptive pathfinding depends on NgCAM and axonin-1, proprioceptive fibers require NrCAM and F11. PMID- 11430806 TI - Retinal ganglion cell genesis requires lakritz, a Zebrafish atonal Homolog. AB - Mutation of the zebrafish lakritz (lak) locus completely eliminates the earliest born retinal cells, the ganglion cells (RGCs). Instead, excess amacrine, bipolar, and Muller glial cells are generated in the mutant. The extra amacrines are found at ectopic locations in the ganglion cell layer. Cone photoreceptors appear unaffected by the mutation. Molecular analysis reveals that lak encodes Ath5, the zebrafish eye-specific ortholog of the Drosophila basic helix-loop-helix transcription factor Atonal. A combined birth-dating and cell marker analysis demonstrates that lak/ath5 is essential for RGC determination during the first wave of neurogenesis in the retina. Our results suggest that this wave is skipped in the mutant, leading to an accumulation of progenitors for inner nuclear layer cells. PMID- 11430807 TI - Homeostatic control of presynaptic release is triggered by postsynaptic membrane depolarization. AB - Homeostatic mechanisms regulate synaptic function to maintain nerve and muscle excitation within reasonable physiological limits. The mechanisms that initiate homeostasic changes to synaptic function are not known. We specifically impaired cellular depolarization by expressing the Kir2.1 potassium channel in Drosophila muscle. In Kir2.1-expressing muscle there is a persistent outward potassium current ( approximately 10 nA), decreased muscle input resistance (50-fold), and a hyperpolarized resting potential. Despite impaired muscle excitability, synaptic depolarization of muscle achieves wild-type levels. A quantal analysis demonstrates that increased presynaptic release (quantal content), without a change in quantal size (mEPSC amplitude), compensates for altered muscle excitation. Because morphological synaptic growth is normal, we conclude that a homeostatic increase in presynaptic release compensates for impaired muscle excitability. These data demonstrate that a monitor of muscle membrane depolarization is sufficient to initiate synaptic homeostatic compensation. PMID- 11430808 TI - Spike-associated fast contraction of dendritic spines in cultured hippocampal neurons. AB - Dendritic spines have long been known to contain contractile elements and have recently been shown to express apparent spontaneous motility. Using high resolution imaging of dendritic spines of green-fluorescent protein (GFP) expressing, patch-clamped hippocampal neurons in dissociated culture, we find that bursts of action potentials, evoked by depolarizing current pulses, cause momentary contractions of dendritic spines. Blocking calcium currents with cobalt prevented these twitches. In additional experiments with neurons loaded via a micropipette with calcium-sensitive and insensitive dyes, spontaneous calcium transients were associated with a rapid contraction of the spine head. The spine twitch was prolonged by tetraethylammonium or bicuculline, which enhance calcium transients, and was blocked by the actin polymerization antagonist latrunculin-B. The spine twitch may be instrumental in modulating reactivity of the NMDA receptor to afferent stimulation, following back-propagating action potentials. PMID- 11430809 TI - Synchronized neural activity in the Drosophila memory centers and its modulation by amnesiac. AB - The mushroom bodies are key features of the brain circuitry for insect associative learning, especially when evoked by olfactory cues. Mushroom bodies are also notable for the close-packed parallel architecture of their many intrinsic neuronal elements, known as Kenyon cells. Here, we report that Kenyon cells of adult Drosophila exhibit synchronous oscillation of intracellular calcium concentration, with a mean period of approximately 4 min. Robust oscillation within a dissected brain persists for hours in insect saline and is strongly modulated in amplitude by the product(s) of the memory consolidation gene, amnesiac. It is also sensitive to pharmacological agents specific for several classes of ion channel and for acetylcholine and GABA receptors. A role in memory consolidation involving transcriptionally mediated synaptic strengthening is proposed. PMID- 11430810 TI - A key role of starburst amacrine cells in originating retinal directional selectivity and optokinetic eye movement. AB - The directional selectivity of retinal ganglion cell responses represents a primitive pattern recognition that operates within a retinal neural circuit. The cellular origin and mechanism of directional selectivity were investigated by selectively eliminating retinal starburst amacrine cells, using immunotoxin mediated cell targeting techniques. Starburst cell ablation in the adult retina abolished not only directional selectivity of ganglion cell responses but also an optokinetic eye reflex derived by stimulus movement. Starburst cells therefore serve as the key element that discriminates the direction of stimulus movement through integrative synaptic transmission and play a pivotal role in information processing that stabilizes image motion. PMID- 11430811 TI - Two-dimensional substructure of MT receptive fields. AB - Neurons at progressively higher levels of the visual system have progressively larger, more complicated receptive fields, presumably constructed from simpler antecedent receptive fields. To study this hierarchical organization, we used sparse white noise to map receptive-field substructure (second order Wiener-like kernels) in an extrastriate motion processing area (MT) of alert monkeys. The maps revealed a clear substructure, on a spatial scale comparable to the receptive fields of the V1 inputs. There were both facilitatory and suppressive interactions that differed in spatial organization and time course. Directional interactions were remarkably precise over a very small spatial range, and reversed when successive stimuli reversed contrast--a neural correlate of "reverse phi" motion perception. The maps of some cells had an unexpected, curved shape, which challenges existing models for direction selectivity. PMID- 11430812 TI - Floccular complex spike response to transparent retinal slip. AB - The flocculus of the rabbit is involved in the plasticity of compensatory eye movements. It is generally assumed that the climbing fiber input to floccular Purkinje cells encodes "retinal slip," which in turn would be a measure for the oculomotor performance error. To test this, we used transparent motion stimuli, creating a retinal slip signal that broke up this relation. We recorded the ensuing oculomotor behavior and complex spike activity of floccular Purkinje cells. Complex spike modulation in response to transparent stimulation was identical to that of a single optokinetic pattern, despite considerably different retinal slip. These results suggest that the climbing fiber code may be effectively related to the eye movement performance error, rather than to retinal slip. PMID- 11430813 TI - Predicting every spike: a model for the responses of visual neurons. AB - In the early visual system, neuronal responses can be extremely precise. Under a wide range of stimuli, cells in the retina and thalamus fire spikes very reproducibly, often with millisecond precision on subsequent stimulus repeats. Here we develop a mathematical description of the firing process that, given the recent visual input, accurately predicts the timing of individual spikes. The formalism is successful in matching the spike trains from retinal ganglion cells in salamander, rabbit, and cat, as well as from lateral geniculate nucleus neurons in cat. It adapts to many different response types, from very precise to highly variable. The accuracy of the model allows a compact description of how these neurons encode the visual stimulus. PMID- 11430814 TI - Dopamine production in the caudate putamen restores feeding in dopamine-deficient mice. AB - Dopamine-deficient (DD) mice cannot synthesize dopamine (DA) in dopaminergic neurons due to selective inactivation of the tyrosine hydroxylase gene in those neurons. These mice become hypoactive and hypophagic and die of starvation by 4 weeks of age. We used gene therapy to ascertain where DA replacement in the brain restores feeding and other behaviors in DD mice. Restoration of DA production within the caudate putamen restores feeding on regular chow and nest-building behavior, whereas restoration of DA production in the nucleus accumbens restores exploratory behavior. Replacement of DA to either region restores preference for sucrose or a palatable diet without fully rescuing coordination or initiation of movement. These data suggest that a fundamental difference exists between feeding for sustenance and the ability to prefer rewarding substances. PMID- 11430815 TI - Effects of attention and emotion on face processing in the human brain: an event related fMRI study. AB - We used event-related fMRI to assess whether brain responses to fearful versus neutral faces are modulated by spatial attention. Subjects performed a demanding matching task for pairs of stimuli at prespecified locations, in the presence of task-irrelevant stimuli at other locations. Faces or houses unpredictably appeared at the relevant or irrelevant locations, while the faces had either fearful or neutral expressions. Activation of fusiform gyri by faces was strongly affected by attentional condition, but the left amygdala response to fearful faces was not. Right fusiform activity was greater for fearful than neutral faces, independently of the attention effect on this region. These results reveal differential influences on face processing from attention and emotion, with the amygdala response to threat-related expressions unaffected by a manipulation of attention that strongly modulates the fusiform response to faces. PMID- 11430816 TI - Yeast [PSI+] "prions" that are crosstransmissible and susceptible beyond a species barrier through a quasi-prion state. AB - The yeast [PSI(+)] element represents an aggregated form of release factor Sup35p and is inherited by a prion mechanism. A "species barrier" prevents crosstransmission of the [PSI(+)] state between heterotypic Sup35p "prions." Kluyveromyces lactis and Yarrowia lipolytica Sup35 proteins, however, show interspecies [PSI(+)] transmissibility and susceptibility and a high spontaneous propagation rate. Cross-seeding was visualized by coaggregation of differential fluorescence probes fused to heterotypic Sup35 proteins. This coaggregation state, referred to as a "quasi-prion" state, can be stably maintained as a heritable [PSI(+)] element composed of heterologous Sup35 proteins. K. lactis Sup35p was capable of forming [PSI(+)] elements not only in S. cerevisiae but in K. lactis. These two Sup35 proteins contain unique multiple imperfect oligopeptide repeats responsible for crosstransmission and high spontaneous propagation of novel [PSI(+)] elements. PMID- 11430817 TI - Cvt19 is a receptor for the cytoplasm-to-vacuole targeting pathway. AB - Cvt19 is specifically required for the transport of resident vacuolar hydrolases that utilize the cytoplasm-to-vacuole targeting (Cvt) pathway. Autophagy (Apg) and pexophagy, processes that use the majority of the same protein components as the Cvt pathway, do not require Cvt19. Cvt19GFP is localized to punctate structures on or near the vacuole surface. Cvt19 is a peripheral membrane protein that binds to the precursor form of the Cvt cargo protein aminopeptidase I (prAPI) and travels to the vacuole with prAPI. These results suggest that Cvt19 is a receptor protein for prAPI that allows for the selective transport of this protein by both the Cvt and Apg pathways. PMID- 11430818 TI - The axial channel of the proteasome core particle is gated by the Rpt2 ATPase and controls both substrate entry and product release. AB - Substrates enter the proteasome core particle (CP) through a channel that opens upon association with the regulatory particle (RP). Using yeast mutants, we show that channel opening is mediated by the ATPase domain of Rpt2, one of six ATPases in the RP. To test whether degradation products exit through this channel, we analyzed their size distribution. Their median length from an open-channel CP mutant was 40% greater than that from the wild-type. Thus, channel opening may enhance the yield of peptides long enough to function in antigen presentation. These experiments demonstrate that gating of the RP channel controls both substrate entry and product release, and is specifically regulated by an ATPase in the base of the RP. PMID- 11430819 TI - Diabetes mellitus and exocrine pancreatic dysfunction in perk-/- mice reveals a role for translational control in secretory cell survival. AB - The protein kinase PERK couples protein folding in the endoplasmic reticulum (ER) to polypeptide biosynthesis by phosphorylating the alpha subunit of eukaryotic translation initiation factor 2 (eIF2alpha), attenuating translation initiation in response to ER stress. PERK is highly expressed in mouse pancreas, an organ active in protein secretion. Under physiological conditions, PERK was partially activated, accounting for much of the phosphorylated eIF2alpha in the pancreas. The exocrine and endocrine pancreas developed normally in Perk-/- mice. Postnatally, ER distention and activation of the ER stress transducer IRE1alpha accompanied increased cell death and led to progressive diabetes mellitus and exocrine pancreatic insufficiency. These findings suggest a special role for translational control in protecting secretory cells from ER stress. PMID- 11430820 TI - Translational control is required for the unfolded protein response and in vivo glucose homeostasis. AB - The accumulation of unfolded protein in the endoplasmic reticulum (ER) attenuates protein synthesis initiation through phosphorylation of the alpha subunit of eukaryotic translation initiation factor 2 (eIF2alpha) at Ser51. Subsequently, transcription of genes encoding adaptive functions including the glucose regulated proteins is induced. We show that eIF2alpha phosphorylation is required for translation attenuation, transcriptional induction, and survival in response to ER stress. Mice with a homozygous mutation at the eIF2alpha phosphorylation site (Ser51Ala) died within 18 hr after birth due to hypoglycemia associated with defective gluconeogenesis. In addition, homozygous mutant embryos and neonates displayed a deficiency in pancreatic beta cells. The results demonstrate that regulation of translation through eIF2alpha phosphorylation is essential for the ER stress response and in vivo glucose homeostasis. PMID- 11430821 TI - An extended RNA binding surface through arrayed S1 and KH domains in transcription factor NusA. AB - The crystal structure of Thermotoga maritima NusA, a transcription factor involved in pausing, termination, and antitermination processes, reveals a four domain, rod-shaped molecule. An N-terminal alpha/beta portion, a five-stranded beta-barrel (S1 domain), and two K-homology (KH) modules create a continuous spine of positive electrostatic potential, suitable for nonspecific mRNA attraction. Homology models suggest how, in addition, specific mRNA regulatory sequences can be recognized by the S1 and KH motifs. An arrangement of multiple S1 and KH domains mediated by highly conserved residues is seen, creating an extended RNA binding surface, a paradigm for other proteins with similar domain arrays. Structural and mutational analyses indicate that the motifs cooperate, modulating strength and specificity of RNA binding. PMID- 11430822 TI - Regulated ARE-mediated mRNA decay in Saccharomyces cerevisiae. AB - The stability of several oncogene, cytokine, and growth factor transcripts is tightly regulated by signaling pathways through an ARE (AU-rich element) present in their 3'-UTRs. We have identified a yeast transcript, TIF51A, whose stability is regulated through its AU-rich 3'-UTR. We demonstrate that the mammalian TNFalpha and c-fos AREs regulate turnover of a reporter yeast transcript in a similar manner. AREs stabilize the transcript in glucose media and function as destabilizing elements in media lacking glucose or when the Hog1p/p38 MAP kinase pathway is inhibited. Significantly, both yeast and mammalian AREs promote deadenylation-dependent decapping in the yeast system. Furthermore, the yeast ELAV homolog, Pub1p, regulates the stability mediated by the TNFalpha ARE. These results demonstrate that yeast possess a regulatable mechanism for ARE-mediated decay and suggest conservation of this turnover process from yeast to humans. PMID- 11430823 TI - Analysis of telomerase processivity: mechanistic similarity to HIV-1 reverse transcriptase and role in telomere maintenance. AB - The key protein subunit of the telomerase complex, known as TERT, possesses a reverse transcriptase (RT)-like domain that is conserved in enzymes encoded by retroviruses and retroelements. Structural and functional analysis of HIV-1 RT suggests that RT processivity is governed, in part, by the conserved motif C, motif E, and a C-terminal domain. Mutations in analogous regions of the yeast TERT were found to have anticipated effects on telomerase processivity in vitro, suggesting a great deal of mechanistic and structural similarity between TERT and retroviral RTs, and a similarity that goes beyond the homologous domain. A close correlation was uncovered between telomerase processivity and telomere length in vivo, suggesting that enzyme processivity is a limiting factor for telomere maintenance. PMID- 11430824 TI - Cdk1 triggers association of RNA polymerase to cell cycle promoters only after recruitment of the mediator by SBF. AB - Activation of HO in yeast involves recruitment of transcription factors in two waves. The first is triggered by inactivation of Cdk1 at the end of mitosis, which promotes import into the nucleus of the Swi5 transcription factor. Swi5 recruits the Swi/Snf chromatin-remodeling complex, which then facilitates recruitment of the SAGA histone acetylase, which in turn permits the binding of the SBF transcription factor. We show here that SBF then recruits the SRB/mediator complex and that this process occurs in the absence of Cdk1 activity. The second wave is triggered by reactivation of Cdk1, which leads to recruitment of PolII, TFIIB, and TFIIH. RNA polymerase is, therefore, recruited to HO in two steps and not as a holoenzyme. A similar sequence of events occurs at other SBF-regulated promoters, such as CLN1, CLN2, and PCL1. PMID- 11430825 TI - Regulation of human flap endonuclease-1 activity by acetylation through the transcriptional coactivator p300. AB - We describe a role for the transcriptional coactivator p300 in DNA metabolism. p300 formed a complex with flap endonuclease-1 (Fen1) and acetylated Fen1 in vitro. Furthermore, Fen1 acetylation was observed in vivo and was enhanced upon UV treatment of human cells. Remarkably, acetylation of the Fen1 C terminus by p300 significantly reduced Fen1's DNA binding and nuclease activity. Proliferating cell nuclear antigen (PCNA) was able to stimulate both acetylated and unacetylated Fen1 activity to the same extent. Our results identify acetylation as a novel regulatory modification of Fen1 and implicate that p300 is not only a component of the chromatin remodeling machinery but might also play a critical role in regulating DNA metabolic events. PMID- 11430826 TI - Role of PML and PML-RARalpha in Mad-mediated transcriptional repression. AB - Fusion of the promyelocytic leukemia (PML) protein to the retinoic acid receptor alpha (RARalpha) generates the transforming protein of acute promyelocytic leukemias. PML appears to be involved in multiple functions, including apoptosis and transcriptional activation by RAR, whereas PML-RARalpha blocks these functions of PML. However, the mechanisms of leukemogenesis by PML-RARalpha remain elusive. Here we show that PML interacts with multiple corepressors (c Ski, N-CoR, and mSin3A) and histone deacetylase 1, and that this interaction is required for transcriptional repression mediated by the tumor suppressor Mad. PML RARalpha has the two corepressor-interacting sites and inhibits Mad-mediated repression, suggesting that aberrant binding of PML-RARalpha to the corepressor complexes may lead to abrogation of the corepressor function. These mechanisms may contribute to events leading to leukemogenesis. PMID- 11430827 TI - Cytoplasmic recruitment of INI1 and PML on incoming HIV preintegration complexes: interference with early steps of viral replication. AB - During the early phase of the retroviral life cycle, only a fraction of internalized virions end up integrating their genome into the chromosome, even though the resulting proviruses are almost systematically expressed. Here, we reveal that incoming retroviral preintegration complexes trigger the exportin mediated cytoplasmic export of the SWI/SNF component INI1 and of the nuclear body constituent PML. We further show that the HIV genome associates with these proteins before nuclear migration. In the presence of arsenic, PML is sequestered in the nucleus, and the efficiency of HIV-mediated transduction is markedly increased. These results unveil a so far unsuspected cellular response that interferes with the early steps of HIV replication. PMID- 11430828 TI - A DNA damage response pathway controlled by Tel1 and the Mre11 complex. AB - We define a DNA damage checkpoint pathway in S. cerevisiae governed by the ATM homolog Tel1 and the Mre11 complex. In mitotic cells, the Tel1-Mre11 complex pathway triggers Rad53 activation and its interaction with Rad9, whereas in meiosis it acts via Rad9 and the Rad53 paralog Mre4/Mek1. Activation of the Tel1 Mre11 complex pathway checkpoint functions appears to depend upon the Mre11 complex as a damage sensor and, at least in meiotic cells, to depend on unprocessed DNA double-strand breaks (DSBs). The DSB repair functions of the Mre11 complex are enhanced by the pathway, suggesting that the complex both initiates and is regulated by the Tel1-dependent DSB signal. These findings demonstrate that the diverse functions of the Mre11 complex in the cellular DNA damage response are conserved in mammals and yeast. PMID- 11430829 TI - The two-handed E box binding zinc finger protein SIP1 downregulates E-cadherin and induces invasion. AB - Transcriptional downregulation of E-cadherin appears to be an important event in the progression of various epithelial tumors. SIP1 (ZEB-2) is a Smad-interacting, multi-zinc finger protein that shows specific DNA binding activity. Here, we report that expression of wild-type but not of mutated SIP1 downregulates mammalian E-cadherin transcription via binding to both conserved E2 boxes of the minimal E-cadherin promoter. SIP1 and Snail bind to partly overlapping promoter sequences and showed similar silencing effects. SIP1 can be induced by TGF-beta treatment and shows high expression in several E-cadherin-negative human carcinoma cell lines. Conditional expression of SIP1 in E-cadherin-positive MDCK cells abrogates E-cadherin-mediated intercellular adhesion and simultaneously induces invasion. SIP1 therefore appears to be a promoter of invasion in malignant epithelial tumors. PMID- 11430830 TI - A hedgehog-insensitive form of patched provides evidence for direct long-range morphogen activity of sonic hedgehog in the neural tube. AB - Cell pattern in the ventral neural tube is organized by Sonic hedgehog (Shh) secreted by floor plate cells. To assay the range of direct Shh action, we developed a general method for blocking transduction of Hedgehog (Hh) signals through ectopic expression of a deleted form of the Hh receptor Patched (Ptc), termed Ptc(Deltaloop2). We validated this method in Drosophila and used mouse Ptc1(Deltaloop2) (mPtc1(Deltaloop2)) to block Shh transduction in the chick neural tube. mPtc1(Deltaloop2) expression caused cell-autonomous ventral-to dorsal switches in progenitor identity and neuronal fate throughout the ventral neural tube, supporting a gradient mechanism whereby Shh acts directly and at long range. mPtc1(Deltaloop2) expression also caused the abnormal spread of Shh to more dorsal cells, indicating that Shh in the neural tube, like Hh in Drosophila, induces a feedback mechanism that limits its range of action. PMID- 11430831 TI - Coupling Met to specific pathways results in distinct developmental outcomes. AB - Receptor tyrosine kinases (RTKs) mediate distinct biological responses by stimulating similar intracellular signaling pathways. Whether the specificity of the response is determined by qualitative or quantitative differences in signaling output is not known. We addressed this question in vivo by replacing the multifunctional docking sites of Met, the receptor for hepatocyte growth factor, with specific binding motifs for phosphatidylinositol-3 kinase, Src tyrosine kinase, or Grb2 (Met(2P), Met(2S), and Met(2G), respectively). All three mutants retained normal signaling through the multiadaptor Gab1, but differentially recruited specific effectors. While Met(2G) mice developed normally, Met(2P) and Met(2S) mice were loss-of-function mutants displaying different phenotypes and rescue of distinct tissues. These data indicate that RTK mediated activation of specific signaling pathways is required to fulfill cell specific functions in vivo. PMID- 11430832 TI - The Peutz-Jegher gene product LKB1 is a mediator of p53-dependent cell death. AB - Here, we investigate the mechanism and function of LKB1, a Ser/Thr kinase mutated in Peutz-Jegher syndrome (PJS). We demonstrate that LKB1 physically associates with p53 and regulates specific p53-dependent apoptosis pathways. LKB1 protein is present in both the cytoplasm and nucleus of living cells and translocates to mitochondria during apoptosis. In vivo, LKB1 is highly upregulated in pyknotic intestinal epithelial cells. In contrast, polyps arising in Peutz-Jegher patients are devoid of LKB1 staining and have reduced numbers of apoptotic cells. We propose that a deficiency in apoptosis is a key factor in the formation of multiple benign intestinal polyps in PJS patients, and possibly for the subsequent development of malignant tumors in these patients. PMID- 11430833 TI - A common phosphate binding site explains the unique substrate specificity of GSK3 and its inactivation by phosphorylation. AB - The inhibition of GSK3 is required for the stimulation of glycogen and protein synthesis by insulin and the specification of cell fate during development. Here, we demonstrate that the insulin-induced inhibition of GSK3 and its unique substrate specificity are explained by the existence of a phosphate binding site in which Arg-96 is critical. Thus, mutation of Arg-96 abolishes the phosphorylation of "primed" glycogen synthase as well as inhibition by PKB mediated phosphorylation of Ser-9. Hence, the phosphorylated N terminus acts as a pseudosubstrate, occupying the same phosphate binding site used by primed substrates. Significantly, this mutation does not affect phosphorylation of "nonprimed" substrates in the Wnt-signaling pathway (Axin and beta-catenin), suggesting new approaches to design more selective GSK3 inhibitors for the treatment of diabetes. PMID- 11430834 TI - Histone H2A.Z acetylation modulates an essential charge patch. AB - Histone H2A.Z is structurally and functionally distinct from the major H2As. To understand the function of H2A.Z acetylation, we performed a mutagenic analysis of the six acetylated lysines in the N-terminal tail of Tetrahymena H2A.Z. Tetrahymena cannot survive with arginines at all six sites. Retention of one acetylatable lysine is sufficient to provide the essential function of H2A.Z acetylation. This essential function can be mimicked by deleting the region encompassing all six sites, or by mutations that reduce the positive charge of the N terminus at the acetylation sites themselves, or at other sites in the tail. These properties argue that the essential function of H2A.Z acetylation is to modify a "charge patch" by reducing the charge of the tail. PMID- 11430835 TI - Topological regulation of cell division in E. coli. spatiotemporal oscillation of MinD requires stimulation of its ATPase by MinE and phospholipid. AB - Topological regulation of cell division in E. coli requires positioning a cell division inhibitor, MinC, at the poles of the cell, thus restricting the potential for division to midcell. This positioning is achieved through a rapid oscillation of MinC from pole to pole, a process requiring MinD and MinE. However, the mechanistic basis for this oscillation is not known. Here we report that MinE stimulates MinD ATPase activity, but only in the presence of phospholipid vesicles. Analysis of MinE mutants demonstrates that this stimulation is required for MinD oscillation and suggests that the level of stimulation determines the period of the oscillation. A model is presented in which the requirements for the MinD ATPase contribute spatial and temporal inputs that provide the mechanistic basis for the oscillation. PMID- 11430836 TI - Unmet needs in diagnosis and treatment of mood disorders in children and adolescents. Proceedings of a consensus development conference. October 17-18, 2000. Washington, DC, USA. PMID- 11430838 TI - Mood disorders in children and adolescents: an NIMH perspective. PMID- 11430839 TI - The treatment of mood disorders in children and adolescents by general pediatricians. PMID- 11430840 TI - Emotional development and psychiatry. AB - This paper discusses the current practice of using single word labels for emotional states that ignore the nature of the incentive, cognitive representations, physiologic profile, and especially the developmental stage of the agent. The universal cognitive advances that are derivatives of brain maturation over the first dozen years are accompanied by significant emotional states that infants and young children cannot experience. The cognitive transitions at the end of the first year, the middle of the second year, 5-7 years and early adolescence have special implications for affect states. PMID- 11430841 TI - Are child-, adolescent-, and adult-onset depression one and the same disorder? AB - This paper reviews prior research studies examining neurobiological correlates and treatment response of depression in children, adolescents, and adults. Although there are some similarities in research findings observed across the life cycle, both children and adolescents have been found to differ from depressed adults on measures of basal cortisol secretion, corticotropin stimulation post-corticotropin releasing hormone (CRH) infusion, response to several serotonergic probes, immunity indices, and efficacy of tricyclic medications. These differences are proposed to be due to 1) developmental factors, 2) stage of illness factors (e.g., number of episodes, total duration of illness), or 3) heterogeneity in clinical outcome (e.g., recurrent unipolar course vs. new-onset bipolar disorder). Relevant clinical and preclinical studies that provide support for these alternate explanations of the discrepant findings are reviewed, and directions for future research are discussed. To determine whether child-, adolescent-, and adult-onset depression represent the same condition, it is recommended that researchers 1) use the same neuroimaging paradigms in child, adolescent, and adult depressed cohorts; 2) carefully characterize subjects' stage of illness; and 3) conduct longitudinal clinical and repeat neurobiological assessments of patients of different ages at various stages of illness. In addition, careful attention to familial subtypes (e.g., depressive spectrum disorders vs. familial pure depressive disorders) and environmental factors (e.g., trauma history) are suggested for future investigations. PMID- 11430842 TI - Mood disorders in children and adolescents: an epidemiologic perspective. AB - Epidemiologic studies show that major depression is comparatively rare among children, but common among adolescents, with up to a 25% lifetime prevalence by the end of adolescence. Mania is much less common, with no more than a 2% lifetime prevalence by the end of adolescence. Developmental studies that include assessments of both hormonal changes and social changes through the pubertal transition are needed to investigate joint biological and environmental influences on the emergence of the gender difference in depression in puberty. Although subthreshold mood disorder symptoms are common, controversy exists about their clinical significance. This controversy is made more complex by methodologic uncertainties regarding inconsistent symptom reports obtained from parents, teachers, and children and by the pervasive existence of comorbidity. Retrospective reports about age of onset in adult studies suggest that at least 50% of youngsters with major depression and 90% of those with mania continue to have adult recurrences. These recurrences are mediated by adverse role transitions, such as truncated educational attainment and teenage childbearing, that typically occur before the time of initial treatment. Aggressive outreach and early treatment aimed at preventing the occurrence of adverse role effects might help decrease the persistence of child and adolescent mood disorders. Long term follow-up studies are needed to resolve current uncertainties regarding nosology, methodology, and long-term treatment effects. Innovative epidemiologic research designs aimed at more quickly providing provisional information are also needed to advance understanding of long-term developmental processes. PMID- 11430843 TI - Ethical issues in research and innovative therapy in children with mood disorders. AB - Research involving children with mood disorders should generally offer a reasonable prospect of benefit to those involved, unless the risks are minimal. While federal regulations require a prospect of direct benefit, from an ethical perspective indirect benefits, such as the advantages of being in a clinical trial, are relevant. Standard care should not be presumed to be preferable to treatment in the context of a clinical trial, particularly if it has not been evaluated in well designed studies involving groups comparable to the patient. Similarly, active treatments in a controlled trial should not be presumed to be preferable to placebos, particularly in studies involving mood disorders where the placebo effect is more likely to be substantial. Innovative therapy-using approved drugs for unapproved purposes-may be more problematic than research, on theoretical and empirical grounds. Studies with no prospect of direct benefit and more than minimal risk are problematic when the patient is incompetent to provide meaningful consent. Whether or not a controlled trial involving children with mood disorders can be ethically justified depends on several factors, including the validity of standard treatment, the seriousness of the disorder, the likelihood of adverse consequences if treatment is delayed, the burden of the proposed intervention, and value judgments by parents, working collaboratively with a caring physician. PMID- 11430844 TI - The role of childhood trauma in the neurobiology of mood and anxiety disorders: preclinical and clinical studies. AB - Epidemiologic studies indicate that children exposed to early adverse experiences are at increased risk for the development of depression, anxiety disorders, or both. Persistent sensitization of central nervous system (CNS) circuits as a consequence of early life stress, which are integrally involved in the regulation of stress and emotion, may represent the underlying biological substrate of an increased vulnerability to subsequent stress as well as to the development of depression and anxiety. A number of preclinical studies suggest that early life stress induces long-lived hyper(re)activity of corticotropin-releasing factor (CRF) systems as well as alterations in other neurotransmitter systems, resulting in increased stress responsiveness. Many of the findings from these preclinical studies are comparable to findings in adult patients with mood and anxiety disorders. Emerging evidence from clinical studies suggests that exposure to early life stress is associated with neurobiological changes in children and adults, which may underlie the increased risk of psychopathology. Current research is focused on strategies to prevent or reverse the detrimental effects of early life stress on the CNS. The identification of the neurobiological substrates of early adverse experience is of paramount importance for the development of novel treatments for children, adolescents, and adults. PMID- 11430845 TI - Genetic and environmental influences on the temporal association between earlier anxiety and later depression in girls. AB - BACKGROUND: The purpose of the present study was to investigate the role of genetic and environmental factors in the association between depressive symptoms and symptoms of overanxious disorder, simple phobias, and separation anxiety in 8 13-year-old and 14-17-year-old girls. METHODS: Multivariate genetic models were fitted to child-reported longitudinal symptom data gathered from clinical interview on 415 MZ [corrected] and 194 DZ [corrected] female twin pairs from the Virginia Twin Study of Adolescent Behavioral Development (VTSABD) [corrected]. RESULTS: Model-fitting results suggest there are distinct etiological [corrected] patterns underlying the association between depression and the different anxiety syndromes during the course of development: 1) specific genetic influences on depression after age 14 reflect liability to symptoms of earlier overanxious disorder (OAD) and simple phobias; 2) aspects of the shared environment that influence symptoms of depression before age 14 contribute to symptoms of separation anxiety and simple phobias later in adolescence [corrected]; 3) the shared environmental influence on [corrected] depression in 14+ girls also affects liability to symptoms of concurrent OAD and persistent separation anxiety. CONCLUSIONS: These results suggest that depression before and after age 14 may be etiologically distinct syndromes. Earlier symptoms of OAD and (to a lesser extent phobic symptoms) [corrected] reflect the same genetic risk, and separation anxiety symptoms both before and after age 14 reflect the same environmental risk that influence liability to depressive symptoms expressed in middle to late adolescence. PMID- 11430846 TI - Diagnosing pediatric depression. AB - This review examines current instrumentation for making clinical and research diagnoses of depressive disorders in children and adolescents. Reliable assessment of depression in children requires gathering information from both the parent and child, as well as from all other available information. The methodology for obtaining information from the child must be adapted to reword and better obtain information in those domains that are inherently difficult for children, including questions about internal affect state and questions requiring judgment. Because child depression is highly comorbid with other psychiatric disorders, including anxiety, attention-deficit/hyperactivity disorder (ADHD), and conduct disorder (CD), it is imperative that these and other psychiatric disorders be simultaneously assessed. A number of structured and semistructured instruments address this task well. More work is needed to decrease the time burden and cost of reliable assessment of child depression. PMID- 11430847 TI - Diagnosis of childhood and adolescent suicidal behavior: unmet needs for suicide prevention. AB - This article highlights strategies for diagnosing risk for childhood and adolescent suicidal behavior. Empirical studies identifying risk factors for childhood and adolescent suicidal behavior guided recommendations for suicide risk assessment. Diagnostic assessment involves identification of multiple factors including demographic characteristics, suicidal behavior, psychopathology, interpersonal problems, family discord, family psychopathology, accessibility of lethal suicide methods, exposure to suicide, and protective factors. Interview methods and self-report questionnaires are reliable and valid in identifying suicidal risk but are limited by low base rates of suicide. Identification of risk factors as foci for intervention is important for suicide prevention. PMID- 11430848 TI - Parsing pediatric bipolar disorder from its associated comorbidity with the disruptive behavior disorders. AB - The unique pattern of comorbidity found in pediatric mania greatly complicates accurate diagnosis, the course of the disorder, and its treatment. The pattern of comorbidity is unique by adult standards, especially its overlap with attention deficit/hyperactivity disorder (ADHD), aggression, and conduct disorder. Clinically, symptoms of mania have been discounted as severe ADHD or ignored in the context of aggressive conduct disorder. This atypicality may lead to neglect of the mood component. The addition of high rates of additional disorders contributes to the severe morbidity, dysfunction, and incapacitation frequently observed in these children. A comprehensive approach to diagnostic evaluation is the keystone to establishing an effective treatment program because response to treatment differs with individual disorders. Recognition of the multiplicity of disorders guides therapeutic options in these often refractory conditions. What was previously considered refractory ADHD, oppositionality, aggression, and conduct disorder may respond after mood stabilization. We review these issues in this article. PMID- 11430849 TI - Comorbidity of depression in children and adolescents: models and evidence from a prospective high-risk family study. AB - Despite abundant research demonstrating the magnitude of comorbidity and its importance in understanding childhood psychopathology, there has been limited empirical research designed to examine the nature and causes of comorbidity among youth. This article reviews the current literature on the magnitude and mechanisms of depressive comorbidity and presents data to exemplify the application of high-risk and longitudinal study designs to investigate patterns and explanations for comorbidity. A prospective family study of offspring at high and low risk for the development of anxiety was used to examine the specificity of familial comorbidity of depression and anxiety and the longitudinal stability of "pure" and comorbid disorders over an 8-year period. Findings suggest some specificity of familial expression, as well as longitudinal specificity, of depression and anxiety. The onset of depression follows the onset of most anxiety subtypes, suggesting the sequential nature of depressive comorbidity. Evaluation of mechanisms for comorbidity is important for the identification of homogeneous syndrome categories that will inform research designed to gain understanding of the pathogenesis of mood or anxiety disorders. PMID- 11430850 TI - Mood disorders in children and adolescents: psychopharmacological treatment. AB - Mood disorders are the leading causes of morbidity and mortality in children and adolescence. As a result, many adolescents are treated with psychopharmacologic agents such as antidepressants and mood stabilizers. To date, research into the safety and efficacy of these medications has lagged behind clinical practice. Several controlled trials of antidepressants in this population have recently been completed or are ongoing, yet few controlled trials of mood stabilizers have been conducted. Although acute efficacy of antidepressants is being addressed, many questions remain about pharmacological treatment of early-onset mood disorders. This article will focus on unmet research needs for the psychopharmacologic treatment of child and adolescent mood disorders. PMID- 11430851 TI - Specific psychotherapies for childhood and adolescent depression. AB - Specific psychotherapies for childhood or adolescent depression have been tested against control or comparative conditions. In school-age children with elevated depressive symptoms, cognitive behavioral therapies (CBT) administered in school settings have proven superior to no treatment or to waiting-list controls in almost all studies. One child study suggests that CBT is superior to alternative psychosocial intervention during acute treatment but not at longer term follow up. No one type of CBT for children has proven more efficacious than others. Studies with adolescents have included subjects with diagnosed depressive disorders, primarily major depression. Seven of nine efficacy studies indicate that CBT is more efficacious than a waiting-list condition or than a non-CBT alternative psychotherapy at the end of acute intervention. Longer term follow-up indicates high rates of remission or recovery among depressed adolescents and no superiority of CBT over other psychotherapies in this regard; however, CBT is associated with more rapid remission of symptoms than is family or supportive therapy. Interpersonal psychotherapy has been demonstrated to be more efficacious than a waiting-list condition or minimal contact clinical management in two acute treatment studies. Research is needed to assess the comparative efficacy of psychotherapeutic interventions, antidepressant medication, and their combination and to develop optimal strategies for facilitating remission and preventing relapse. PMID- 11430852 TI - Prevention of childhood depression: recent findings and future prospects. AB - Traditionally, research on childhood mood disorders has focused on clinical trials and longitudinal course and outcome studies, rather than on prevention. Recently, however, advances in the design, methodology, and evaluation of prevention approaches and progress in understanding what factors predispose children to depression have made possible the development of theoretically driven, empirically justified approaches to the prevention of depression in youngsters who are at high risk, either because of elevated symptom levels or parental mood disorder. In this review, we outline recent empirical findings on risk factors for depression in nonreferred samples of youngsters and also in children of depressed parents. Additionally, we review three trials of preventive interventions for childhood depression that yield promising initial findings. We emphasize the need to understand both risks for depression and factors that protect youngsters at risk from succumbing to depression in guiding the development of prevention programs. We also argue that consideration of prevention of depression requires addressing broader social adversity influences that lead to poor mental health outcomes in children, even beyond the effects of parental mood disorder. We conclude with an emphasis on the importance of a developmental-transactional perspective that highlights opportunities for intervention at different points across the lifespan. PMID- 11430853 TI - Affective disorders in children and adolescents: addressing unmet need in primary care settings. AB - Affective disorders are common among children and adolescents but may often remain untreated. Primary care providers could help fill this gap because most children have primary care. Yet rates of detection and treatment for mental disorders generally are low in general health settings, owing to multiple child and family, clinician, practice, and healthcare system factors. Potential solutions may involve 1) more systematic implementation of programs that offer coverage for uninsured children; 2) tougher parity laws that offer equity in defined benefits and application of managed care strategies across physical and mental disorders; and 3) widespread implementation of quality improvement programs within primary care settings that enhance specialty/primary care collaboration, support use of care managers to coordinate care, and provide clinician training in clinically and developmentally appropriate principles of care for affective disorders. Research is needed to support development of these solutions and evaluation of their impacts. PMID- 11430854 TI - Services and prevention: pharmacoepidemiology of antidepressant use. AB - Between 1988 and 1994, data from 3 large sites revealed a 3-5 fold increase in the prevalence of antidepressant (ATD) treatment for U.S. youths aged 2-19 years. In 1994, the ATD prevalence for youths of this age ranged from 13 per 1000 (in the HMO) to 18 per 1000 (in 2 state Medicaid systems). Males predominated in the 10-14-year-olds treated with ATDs, whereas females predominated among 15-19-year olds. Caucasians were more than twice as likely to receive ATD therapy than their African-American counterparts. Primary care providers were the major source of ATD prescriptions for youths. The leading diagnoses in primary care were ADHD followed by depression, whereas the diagnostic order was reversed for youths who received psychiatric services. This review provides details concerning these patterns and trends in ATD treatment of youths from community-based clinical data sources. In addition, the role of these data in an expanded, comprehensive psychotropic knowledge base is discussed. Finally, the implications of an expanded knowledge base for ATD treatments are discussed in regard to generating research questions on effectiveness and safety and to improve treatment consensus within a public-health perspective. PMID- 11430855 TI - Service system finance: implications for children with depression and manic depression. AB - An estimated 6.2% of children in the United States satisfy the criteria for a depression diagnosis, but approximately half of this group do not receive necessary treatment. Thus it is important to consider potential barriers to use through service system finance. This article reviews three major types of changes affecting access: parity legislation, managed care, and public contracting. How these developments will affect children with depression and manic depression (DMD) is unclear. To better understand the potential effects on children with DMD, this review uses new data from the Medical Expenditure Panel Survey to describe the service use patterns of this population. These children have higher levels of expenditures, higher rates of inpatient use, and higher rates of Medicaid payment than do other children with mental health diagnoses; they also are overrepresented among the costliest cases of mental illness in children. Children with DMD pay a relatively low out-of-pocket share, suggesting that parity efforts focusing only on copayments and deductibles will have little effect on the absolute out-of-pocket burden for these children. Because children with DMD are overrepresented among high utilizers of health services, health care rationing arrangements or techniques, such as utilization review and capitation, may place this population at particular risk. PMID- 11430856 TI - Effect of neonatal capsaicin treatment on neural activity in the medullary dorsal horn of neonatal rats evoked by electrical stimulation to the trigeminal afferents: an optical, electrophysiological, and quantitative study. AB - To elucidate which glutamate receptors, NMDA or non-NMDA, have the main role in synaptic transmission via unmyelinated afferents in the trigeminal subnucleus caudalis (the medullary dorsal horn), and to examine the early functional effects of neonatal capsaicin treatment to the subnucleus caudalis, optical recording, field potential recording, and quantitative study using electron micrographs were employed. A medulla oblongata isolated from a rat 5--7 days old was sectioned horizontally 400-microm thick or parasagittally and stained with a voltage sensitive dye, RH482 or RH795. Single-pulse stimulation with high intensity to the trigeminal afferents evoked optical responses mainly in the subnucleus caudalis. The optical signals were composed of two phases, a fast component followed by a long-lasting component. The spatiotemporal properties of the optical signals were well correlated to those of the field potentials recorded simultaneously. The fast component was eliminated by 6-cyano-7-nitro-quinoxaline 2,3-dione (CNQX; 10 microM), while the long-lasting component was not. The latter increased in amplitude under a condition of low Mg(2+) but was significantly reduced by DL-2-amino-5-phosphonovaleric acid (AP5; 30 microM). Neonatal capsaicin treatment also reduced the long-lasting component markedly. In addition, the decreases in the ratio of unmyelinated axons to myelinated axons and in the ratio of unmyelinated axons to Schwann cell subunits of trigeminal nerve roots both showed significant differences (P<0.05, Student's t-test) between the control group and the neonatal capsaicin treatment group. This line of evidence indirectly suggests that synaptic transmission via unmyelinated afferents in the subnucleus caudalis is mediated substantially by NMDA glutamate receptors and documented that neonatal capsaicin treatment induced a functional alteration of the neural transmission in the subnucleus caudalis as well as a morphological alteration of primary afferents within several days after the treatment. PMID- 11430857 TI - Quantitative autoradiographic mapping of the ORL1, mu-, delta- and kappa receptors in the brains of knockout mice lacking the ORL1 receptor gene. AB - Until recently the opioid receptor family was thought to consist of only the mu-, delta- and kappa-receptors. The cloning of opioid receptor like receptor (ORL1) and its endogenous ligand nociceptin/orphanin FQ, which displayed anti-opioid properties, has raised the issue of functional co-operativity of this system with the classical opioid system. ORL1 receptor knockout mice have been successfully developed by homologous recombination to allow the issue of potential heterogeneity of this receptor and also of compensatory changes in mu-, delta- or kappa-receptors in the absence of ORL1 to be addressed. We have carried out quantitative autoradiographic mapping of these receptors in the brains of mice that are wild-type, heterozygous and homozygous for the deletion of the ORL1 receptor. ORL1, mu-, delta- and kappa-receptors were labelled with [(3)H] leucyl nociceptin (0.4 nM), [(3)H] DAMGO (4 nM), [(3)H] deltorphin-I (7 nM), and [(3)H] CI-977 (2.5 nM) respectively. An approximately 50% decrease in [(3)H] leucyl nociceptin binding was seen in heterozygous ORL1 mutant mice and there was a complete absence of binding in homozygous brains indicating the single gene encodes for the ORL1 receptor and any putative subtypes. No significant gross changes in the binding to other opioid receptors were seen across genotypes in the ORL1 mutant mice demonstrating a lack of major compensation of classical opioid receptors in the absence of ORL1. There were a small number of region specific changes in the expression of classical opioid receptors that may relate to interdependent function with ORL1. PMID- 11430858 TI - Lesion of the insular cortex affects luteinizing hormone and testosterone secretion of rat. Lateralized effect. AB - The possible involvement of the insular cortex in the neural control of the hypophyseal-testicular axis was studied in male rats. Right- but not left-sided lesion of the insular cortex resulted in a significant decrease in basal testosterone secretion in vitro and serum testosterone concentration. Both right- and left-sided lesions of the insular cortex induced significant increase in serum luteinizing hormone (LH) concentration. Unilateral lesion of the insular cortex on either sides had no effect on serum follicle stimulating hormone (FSH) level. The results indicate that the insular cortex is involved in the control of testosterone and LH secretion. The data further suggest that the right insular cortex plays a predominant role in the control of male endocrine reproductive processes. PMID- 11430859 TI - Dendritic structure of single hippocampal neurons according to sex and hemisphere of origin in middle-aged and elderly human subjects. AB - The organization of basilar dendritic patterns in the CA1 hippocampal region obtained from 13 middle-aged and elderly human subjects was assessed using the Golgi method. Neurons were classified according to hemisphere of origin and the sex of the respective subjects. Three parameters were measured: total dendritic length (TDL), number of dendritic segments (NDS) and average segment length (ASL, which is TDL divided by NDS). Dendritic segments were classified into proximal (first to third order) and distal (fourth order and above). Sex differences were found in distal TDL and in proximal and distal NDS, neurons belonging to males having larger values than those belonging to females. In addition, a hemispheric difference was detected in distal TDL, in which neurons of the left hemisphere had larger values than those of the right hemisphere. PMID- 11430860 TI - The release of noradrenaline in the locus coeruleus and prefrontal cortex studied with dual-probe microdialysis. AB - The present study was undertaken to investigate and compare the properties of noradrenaline release in the locus coeruleus (LC) and prefrontal cortex (PFC). For that aim the dual-probe microdialysis technique was applied for simultaneous detection of noradrenaline levels in the LC and PFC in conscious rats. Calcium omission in the LC decreased noradrenaline levels in the LC, but increased its levels in the PFC. Novelty increased noradrenaline levels in both structures. Infusion of the alpha(2)-adrenoceptor agonist clonidine decreased extracellular noradrenaline in the LC as well as in the PFC. Infusion of the alpha(2A) adrenoceptor antagonist BRL44408, or the alpha(1)-adrenoceptor agonist cirazoline into the LC or PFC caused a similar dose-dependent increase in both structures. When BRL44408 or cirazoline were infused into the LC, few effects were seen in the PFC. Infusion of the 5-HT(1A)-receptor agonist flesinoxan into the LC or the PFC decreased the release of noradrenaline in both structures. When flesinoxan was infused into the LC, no effects were seen in the PFC. When the GABA(A) antagonist bicuculline was applied to the LC, noradrenaline increased in the LC as well as in the PFC. It is concluded that the release of noradrenaline from somatodendritic sites and nerve terminals responded in a similar manner to presynaptic receptor modulation. The possible existence of dendritic noradrenaline release is discussed. PMID- 11430861 TI - Interaction between the cholinergic system and CRH in the modulation of spatial discrimination learning in mice. AB - Both cholinergic and CRH systems have been linked to cognitive processes such as learning and memory, and neuroanatomical as well as neurochemical evidence suggests important interactions between these two systems. Moreover, recent reports of pro-mnestic effects of CRH open the possibility that CRH could have beneficial effects in animals with cholinergic dysfunction. In a first experiment, spatial discrimination of C57BL/6 mice treated with various doses of scopolamine (0.5--2.0 mg/kg IP) was tested in a two-choice water maze task. Scopolamine, but not methylscopolamine, impaired accuracy and decreased responsivity. In contrast, similar doses of the nicotinic antagonist mecamylamine had no effect on choice accuracy but altered responsivity, as indicated by increased errors of omission and a reduction in swim speed during early experimental stages. ICV CRH (0.5--1.0 microg) also failed to significantly affect accuracy, but a strong tendency was observed to impair percentage correct responses. Measures of responsivity, such as errors of omission, choice latency and distance traveled, and of thigmotaxis were not significantly affected by CRH. However, initial swim speed was reduced by the peptide. Combined treatment with scopolamine (0.5 mg/kg IP) and CRH (0.5 microg ICV) had only mild, and primarily independent, effects, but overall suggested that concomitant blockade of muscarinic receptors and activation of the CRH system would rather act synergistically to disrupt spatial discrimination learning. Synergistic effects were also observed when animals receiving a combination of mecamylamine (2.0 mg/kg IP) and CRH (0.5 microg ICV) were tested, both in terms of responsivity and thigmotaxis, and there was limited evidence that part of these effects were potentiating. Thus, the cholinergic and CRH systems interact in the modulation of learning, but CRH, contrary to prediction, worsens the impairment caused by cholinergic blockade. PMID- 11430862 TI - Differential impact of predator or immobilization stressors on central corticotropin-releasing hormone and bombesin-like peptides in Fast and Slow seizing rat. AB - Lines of rats selectively bred for amygdala excitability, as reflected by kindling rates in response to electrical stimulation, also exhibit differences in tests of anxiety. Inasmuch as corticotropin-releasing hormone (CRH) and bombesin (BN) have been associated with anxiety, regional levels and release of these peptides, as well as plasma adrenocorticotropic hormone (ACTH) and corticosterone, were assessed in 'Slow' and 'Fast' seizing rats following predator exposure (ferret) or immobilization. Ferret exposure elicited a greater increase of plasma ACTH and corticosterone concentrations in the Slow than in the Fast rats. In contrast, immobilization provoked a greater rise of plasma ACTH levels in the Fast rats, paralleling the vigorous struggling observed in this line. In Slow rats, stressor exposure elicited increased levels of ir-BN at the anterior hypothalamus, and increased ir-CRH at the median eminence/arcuate nucleus (Me/Arc), paraventricular hypothalamic nucleus (PVN) and pituitary (Pit), whereas decreased levels of ir-BN were found at the nucleus tractus solitarius (NTS). Fast rats likewise showed decreased ir-BN at the NTS, but unlike the Slow rats, ir-CRH was reduced in the Me/Arc, PVN and Pit in response to both stressors. In vivo microdialysis experiments revealed that in response to ferret exposure, the Slow rats showed a greater CRH release at the central nucleus of the amygdala (CeA) as compared to Fast rats. However, immobilization elicited a more pronounced release of CRH in Fast than in Slow rats. Taken together, the results demonstrate that these two lines of rats show differential endocrinological and neurochemical response patterns to these stressors. PMID- 11430863 TI - Spreading depression in human neocortical slices. AB - Cortical spreading depression (CSD) occurrence has been suggested to be associated with seizures, migraine aura, head injury and brain ischemia infarction. Only few studies identified CSD in human neocortical slices and no comprehensive study so far evaluated this phenomenon in human. Using the neocortical tissue excised for treatment of intractable epilepsy, we aimed to investigate CSD in human. CSD was induced by KCl injection and by modulating T type Ca(2+) currents in incubated human neocortical tissues in an interphase mode. The DC-fluctuations were recorded by inserting microelectrodes into different cortical layers. Local injection of KCl triggered single CSD that propagated at 3.1+/-0.1 mm/min. Repetitive CSD also occurred spontaneously during long lasting application (5 h) of the T-type Ca(2+) channel blockers amiloride (50 microM) or NiCl(2) (10 microM) which was concomitant with a reversible extracellular potassium increase up to 50 mM. CSD could be blocked by the N methyl-D-aspartate receptor antagonist 2-amino-5-phosphonovaleric acid in all cases. The results demonstrate that modulation of the Ca(2+) dynamics conditioned human neocortical slices and increased their susceptibility to generate CSD. Furthermore, these data indicate that glutamatergic pathway plays a role in CSD phenomenon in human. PMID- 11430864 TI - gamma-Aminobutyric acid receptor subtype antagonists differentially alter opioid induced feeding in the shell region of the nucleus accumbens in rats. AB - Food intake is significantly increased by administration of mu-selective opioid agonists into the nucleus accumbens, particularly its shell region. Pretreatment with either opioid (mu, delta(1), delta(2) or kappa(1)) or dopaminergic (D(1)) receptor antagonists in the nucleus accumbens shell reduce mu opioid agonist induced feeding. Selective GABA(A) (muscimol) and GABA(B) (baclofen) agonists administered into the nucleus accumbens shell each stimulate feeding which is respectively and selectively blocked by GABA(A) (bicuculline) and GABA(B) (saclofen) antagonists. The present study investigated whether feeding elicited by the mu-selective opioid agonist, [D-Ala(2),NMe(4),Gly-ol(5)]-enkephalin in the nucleus accumbens shell was decreased by intra-accumbens pretreatment with an equimolar dose range of either GABA(A) or GABA(B) antagonists, and further, whether general opioid or selective GABA antagonists decreased feeding elicited by GABA(A) or GABA(B) agonists in the nucleus accumbens shell. Feeding elicited by the mu-selective opioid agonist was dose-dependently increased following intra accumbens pretreatment with GABA(A) (bicuculline) antagonism; this enhancement was significantly blocked by pretreatment with general or mu-selective opioid antagonists. In contrast, mu opioid agonist-induced feeding elicited from the nucleus accumbens shell was dose-dependently decreased by GABA(B) (saclofen) antagonism. Neither bicuculline nor saclofen in the nucleus accumbens shell altered baseline food intake. Whereas muscimol-induced feeding elicited from the nucleus accumbens shell was reduced by bicuculline and naltrexone, but not saclofen pretreatment, baclofen-induced feeding elicited from the nucleus accumbens shell was reduced by saclofen, but not by bicuculline or naltrexone. These data indicate that GABA(A) and GABA(B) receptor subtype antagonists differentially affect feeding elicited by mu opioid receptor agonists within the nucleus accumbens shell in rats. PMID- 11430865 TI - Motoric behavior in aged rats treated with GM1. AB - Aging is associated with impaired motor function. Nigrostriatal dopaminergic neurons, in part, regulate motoric behavior, and undergo degenerative changes during aging. GM1 ganglioside partially restores pre-synaptic dopaminergic markers and the number and morphology of dopaminergic neurons in the midbrain and striatum of Sprague--Dawley aged rats. These studies investigated whether GM1 treatment, 30 mg/kg, i.p. daily for 36 days, affects locomotor and stereotypic activity, as well as coordination, balance, and strength in aged rats. Under the treatment conditions used, GM1 did not improve the reduced locomotor and stereotypic behavior of the aged rats. While it partially improved performance on a square bridge test, GM1 had no effect on inclined screen and rod suspension tests. Although GM1 restored the decreased content of dopamine and homovanillic acid in the nigrostriatal neurons of the aged rats, it had no effect on the reduced D1 and D2 dopamine receptor binding and mRNA in the striatum. It appears, that despite the morphological and metabolic restoration of aged nigrostriatal neurons, GM1 has limited ability in improving age-associated motor deficits. PMID- 11430866 TI - Role of Na(+)/H(+) exchanger in dilator responses of rat basilar artery in vivo. AB - We tested the hypothesis that activation of Na(+)/H(+) exchanger is involved in dilator responses of the basilar artery to endothelium-dependent vasodilators in vivo. Using a cranial window in anesthetized rats, we examined responses of the basilar artery to acetylcholine and bradykinin. Topical application of acetylcholine and bradykinin increased diameter of the basilar artery in a concentration-related manner. Because N(G)-nitro-L-arginine, an inhibitor of nitric oxide synthase, almost abolished vasodilator responses to acetylcholine and bradykinin, vasodilatation produced by the agonists appears to be mediated primarily by nitric oxide. 5-N,N-Hexamethyleneamiloride, an inhibitor of Na(+)/H(+) exchanger, did not affect baseline diameter of the basilar artery, but inhibited vasodilatation in response to acetylcholine and bradykinin, without affecting vasodilatation produced by sodium nitroprusside. FR183998, another inhibitor of Na(+)/H(+) exchanger, also attenuated acetylcholine-induced dilatation of the basilar artery without affecting vasodilatation in response to sodium nitroprusside. Monomethylamine hydrochloride, which produces intracellular alkalinization, enhanced acetylcholine-induced dilatation of the basilar artery in the presence of 5-N,N-hexamethyleneamiloride. These results suggest that intracellular alkalinization produced by activation of Na(+)/H(+) exchanger may enhance nitric oxide production in the basilar arterial endothelium and thereby contribute to dilator responses of the artery in vivo. PMID- 11430867 TI - Impairment in motor learning of somatostatin null mutant mice. AB - Somatostatin was first identified as a hypothalamic factor which inhibits the release of growth hormone from the anterior pituitary (somatotropin release inhibitory factor, SRIF). Both SRIF and its receptors were subsequently found widely distributed within and outside the nervous system, in the adult as well as in the developing organism. Reflecting this wide distribution, somatostatin has been implicated regulating a diverse array of biological processes. These include body growth, homeostasis, sensory perception, autonomous functions, rate of intestinal absorption, behavior, including cognition and memory, and developmental processes. We produced null mutant mice lacking somatostatin through targeted mutagenesis. The mutant mice are healthy, fertile, and superficially indistinguishable from their heterozygous and wildtype littermates. A 'first round' phenotype screen revealed that mice lacking somatostatin have elevated plasma growth hormone levels, despite normal body size, and have elevated basal plasma corticosterone levels. In order to uncover subtle and unexpected differences, we carried out a systematic behavioral phenotype screen which identified a significant impairment in motor learning revealed when increased demands were made on motor coordination. Motor coordination and motor learning require an intact cerebellum. While somatostatin is virtually absent from the adult cerebellum, the ligand and its receptor(s) are transiently expressed at high levels in the developing cerebellum. This result suggests the functional significance of transient expression of SRIF and its receptors in the development of the cerebellum. PMID- 11430868 TI - Rapid activation of microglial cells by hypoxia, kainic acid, and potassium ions in slice preparations of the rat hippocampus. AB - Microglial activation induced by hypoxia, kainic acid and elevated potassium concentration, all of which alter neuronal function, was studied in hippocampal slices. The activation of microglia was detected by immunostaining with a monoclonal antibody (OX-42) raised against a type 3 complement receptor (CD11b). During activation the phenotype of microglia changes and the intensity of staining of individual cells increases. Oxygen deprivation depressed the focal responses of CA1 neurons to stratum radiatum volleys. Microglial activation was time dependent. Ten minute hypoxia caused mild activation, and after 20 min, a strong microglial reaction could be observed. Although neuronal function returned during reoxygenation, the morphological signs of microglial activation remained. Epileptiform activity of hippocampal neurons, followed by depression, was induced by application of 0.5 mM kainic acid, in a time and dose dependent manner. Washing out kainic acid did not alter microglial reaction. Elevated concentrations of potassium ions induced microglial changes similar to those induced by hypoxia and kainic acid. It is therefore suggested that an elevated extracellular potassium ion concentration may be the common factor in microglial activation observed in these experiments since this is raised both in hypoxia and under the effect of excitotoxins. PMID- 11430869 TI - Nicotine produces selective degeneration in the medial habenula and fasciculus retroflexus. AB - Nicotine's neurotoxic properties in rats were investigated by administering (-) nicotine tartrate for 5 days either continuously in doses of 5.01, 5.72, 6.44, 7.13, 20.41 and 43.1 mg/kg/day via osmotic minipump or intermittently at 11.32 mg/kg/day via one daily subcutaneous injection. As assessed by silver staining, neurotoxicity was seen almost exclusively in the axons of the medial habenula and its output tract, the fasciculus retroflexus, in all treatment groups except the lowest dose. Within the habenula, the damage was noted in the ventral-medial-most portion of the nucleus which is thought to be dense with the alpha 4 beta 2 and/or alpha 3 beta 4 receptor subtypes. Past research has shown the medial habenula to be highly sensitive to the effects of nicotine, and these findings, in conjunction with related research using dopaminergic stimulants, indicate that the habenula may be a weak link in the neurotoxicity seen following stimulant drugs of abuse. PMID- 11430870 TI - Carrier-mediated processes in blood--brain barrier penetration and neural uptake of paraquat. AB - Due to the structural similarity to N-methyl-4-phenyl pyridinium (MPP(+)), paraquat might induce dopaminergic toxicity in the brain. However, its blood- brain barrier (BBB) penetration has not been well documented. We studied the manner of BBB penetration and neural cell uptake of paraquat using a brain microdialysis technique with HPLC/UV detection in rats. After subcutaneous administration, paraquat appeared dose-dependently in the dialysate. In contrast, MPP(+) could not penetrate the BBB in either control or paraquat pre-treated rats. These data indicated that the penetration of paraquat into the brain would be mediated by a specific carrier process, not resulting from the destruction of BBB function by paraquat itself or a paraquat radical. To examine whether paraquat was carried across the BBB by a certain amino acid transporter, L-valine or L-lysine was pre-administered as a co-substrate. The pre-treatment of L valine, which is a high affinity substrate for the neutral amino acid transporter, markedly reduced the BBB penetration of paraquat. When paraquat was administered to the striatum through a microdialysis probe, a significant amount of paraquat was detected in the striatal cells after a sequential 180-min washout with Ringer's solution. This uptake was significantly inhibited by a low Na(+) condition, but not by treatment with putrescine, a potent uptake inhibitor of paraquat into lung tissue. These findings indicated that paraquat is possibly taken up into the brain by the neutral amino acid transport system, then transported into striatal, possibly neuronal, cells in a Na(+)-dependent manner. PMID- 11430871 TI - Localisation of the SRY-related HMG box protein, SOX9, in rodent brain. AB - Human mutations in the transcription factor gene, SOX9, cause campomelic dysplasia (CD), a severe dwarfism associated with brain abnormalities including dilation of lateral ventricles, hypoplasia of the corpus callosum and cerebellum defects. To improve our understanding of how SOX9 contributes to the molecular genetic pathway of brain development we sought to investigate the distribution of SOX9 protein in rat and mouse brain. The regions of SOX9 expression identified in this study correlated with the sites of reported brain abnormalities in CD patients. SOX9 immunoreactivity was observed in nuclei of scattered cells throughout the brain, in the ependymal layer and cells of the choroid plexus. In the forebrain most SOX9-immunoreactive nuclei co-localised with the glial astrocyte marker S-100. In the cerebellum, SOX9 was observed mostly in cells surrounding Purkinje cells, which were identified, by electron microscopy, as Golgi epithelial cells, also known as Bergmann glia. Using SOX9 antibody as a marker for the precursors of Bergmann glia, we traced their origin during mouse development. At embryonic day (E)14.5 and E16.5, SOX9 immunoreactivity was present mainly in the primordial choroid plexus, and ventricular zone. By E18.5, SOX9 was observed in the granular cell and Purkinje cell layers but no labelling was detectable in the external granular layer. These results suggest that SOX9 immunoreactivity is a marker for Bergmann cells during development and favour the proposed origin of the secondary glial scaffold arising from Bergmann cells derived exclusively from the ventricular zone. PMID- 11430872 TI - Spatial disparity sensitivity in area PMLS of the Siamese cat. AB - Previous studies of the visual system of Siamese cats have shown that binocular cells are scarce in areas 17, 18 and 19, yet significantly more abundant in suprasylvian areas such as the postero-medial lateral suprasylvian area (PMLS). The present study aims at evaluating the sensitivity to spatial disparity of PMLS binocular cells in paralyzed and anesthetized Siamese cats. Centrally located receptive fields were mapped, separated using prisms and then stimulated simultaneously using two luminous bars optimally adjusted to the size of the excitatory receptive fields. Delays were introduced in the arrival of the luminous bars in the receptive fields so as to create the desired spatial disparities. Results indicate that approximately a third of PMLS units are binocular and that these binocular cells can detect spatial disparity cues. Indeed, although the sample was relatively small, cells of the tuned excitatory (14/34), tuned inhibitory (2/34), near (6/34) and far (1/34) types were identified. The spatial selectivity, as measured by the width at half height of the tuning curves of the excitatory and inhibitory cells and the slopes of the near and far cells, was similar to that obtained in PMLS of normal cats but not as precise as that found for primary visual areas in these animals. This suggests that these cells might serve as a substrate for coarse stereopsis. PMID- 11430873 TI - Diphenyl diselenide and diphenyl ditelluride affect the rat glutamatergic system in vitro and in vivo. AB - The aim of this study was to investigate the possible involvement of the glutamatergic system in the toxicity of organochalcogens, since this is an important neurotransmitter system for signal transduction and neural function. The results indicated that 100 microM diphenyl diselenide (PhSe)(2) and diphenyl ditelluride (PhTe)(2) inhibit by 50 and 70% (P<0.05), respectively, [(3)H]glutamate binding in vitro. Acute administration of 25 micromol/kg (PhSe)(2) or 3 micromol/kg (PhTe)(2) caused a significant reduction in [(3)H]glutamate (30%, P<0.05) or [(3)H]MK-801 binding (30%, P<0.05) to rat synaptic membranes. These results suggest that (PhSe)(2) and (PhTe)(2) affect, in a rather complex way, the glutamatergic system after acute in vivo exposure in rats. In vitro, total [(3)H]GMP-PNP binding was inhibited about 40% at 100 microM (PhSe)(2) and (PhTe)(2). Acute exposure in vivo to (PhSe)(2) decreased the stable [(3)H]GMP-PNP binding to 25% and (PhTe)(2) to 68% of the control value (P<0.05, for both compounds). Simultaneously, the unstable binding of [(3)H]GMP-PNP was decreased about 30 and 50% (P<0.05, for both compounds) after exposure to (PhSe)(2) and (PhTe)(2), respectively. GMP-PNP stimulated adenylate cyclase (AC) activity significantly in control animals. (PhSe)(2)- and (PhTe)(2)-treated animals increased the basal activity of this enzyme, but GMP-PNP stimulation was totally abolished. These results suggest that the toxic effects of organochalcogens could result from action at different levels of neural signal transduction pathways, possibly involving other neurotransmitters besides the glutamatergic system. PMID- 11430874 TI - Contribution of central amiloride-sensitive transport systems to the development of hypertension in spontaneously hypertensive rats. AB - This study was conducted to examine if central amiloride-sensitive transport systems are involved in the development and/or maintenance of hypertension in spontaneously hypertensive rats (SHR). Either amiloride (75 microg/60 microl/day) or artificial cerebrospinal fluid (aCSF, 60 microl/day) was infused centrally (i.c.v.) for 4 weeks to development (4-5-weeks-old) and maintenance (10-12-weeks old) phases of hypertension in SHR. In development phase, amiloride i.c.v. (n=14) blunted the elevation of blood pressure (BP) compared to aCSF i.c.v. (n=9) (amiloride vs. aCSF; after 3 weeks of i.c.v., 146+/-3 vs. 166+/-5 mmHg, P<0.001). The difference of BP at 3 weeks of i.c.v. was canceled after ganglionic block with hexamethonium (115+/-4 vs. 117+/-5 mmHg). Further, pressor responsiveness to norepinephrine was augmented in amiloride i.c.v. rats (amiloride, n=11 vs. aCSF, n=6; %Delta BP at 800 ng/kg/min.: 16.9+/-1.3 vs. 10.8+/-1.4 mmHg, P<0.05) and this augmentation disappeared after ganglionic block. Pressor responsiveness to angiotensin II and cumulative sodium balance did not differ in the two groups. Intravenous administration of amiloride at the same dose did not attenuate the development of hypertension. On the other hand, in maintenance phase, amiloride i.c.v. by the same protocol as in development phase had no effect on BP in SHR. Also, amiloride i.c.v. did not affect BP in normotensive Wistar-Kyoto rats. These results suggest that central amiloride-sensitive transport systems are involved in the development, but not in the maintenance, of hypertension in SHR through the modulation of autonomic neural mechanisms. PMID- 11430875 TI - Release of dopamine by perfusion with 1-methyl-4-phenylpyridinium ion (MPP(+)) into the striatum is associated with hydroxyl free radical generation. AB - In Parkinson's disease (PD), the dopamine (DA) neuronal cell death in the nigrostriatal system has been proposed to be mediated by reactive oxygen radicals such as hydroxyl radicals (.OH). This.OH production may cause lipid peroxidation of cell membranes leading to neuronal cell death. This paper report that the DA selective neurotoxin, 1-methyl-4-phenylpyridinium ion (MPP(+)), (1 nmol/microl per min for 1 h) infusion into the striatum of rats induces elevation of extracellular DA and.OH formation. These elevations seem to induce lipid peroxidation of striatum membranes, as detected by increases in non-enzymatic formation of 2,3-dihydroxybenzoic acid (DHBA) levels. To test the involvement of DA release in the.OH generation and lipid peroxidation, the rats were pretreated with reserpine (5 mg/kg, i.v., 24 h before MPP(+) or without MPP(+)) to deplete presynaptic DA. Reserpine treatment alone did not change the levels of DA or 2,3 DHBA, while the combined treatment with both MPP(+) and reserpine clearly decreased 2,3-DHBA, as well as DA levels, compared to those in the group treated with MPP(+) alone. After injection into reserpinized rats, DA at various doses (2, 5 and 10 microM) small increased 2,3-DHBA levels dose-dependently, as compared to the MPP(+) alone-treated group. These results clearly indicate that MPP(+) perfusion into the striatum increases extracellular DA levels and this increase may concomitantly induce the formation of reactive free oxygen radicals, such as.OH free radicals. These events may contribute, at least in part, to the nigrostriatal neurons cell death after MPP(+). PMID- 11430876 TI - Intergeniculate leaflets lesion delays but does not prevent the integration of photoperiodic change by the suprachiasmatic nuclei. AB - The duration of the photosensitive phase of the suprachiasmatic nuclei (SCN), as revealed by light-induced Fos protein expression, depends on the photoperiod and is tied to the length of the night. We show here in Syrian hamsters that after a transfer from long to short photoperiod, lengthening of the photosensitive phase of the SCN is significantly delayed but not abolished when the intergeniculate leaflets (IGL) are lesioned. Thus IGL modulate the integration by the SCN of a photoperiodic change. PMID- 11430877 TI - Effects of histamine H3-receptor ligands on brain monoamine oxidase in various mammalian species. AB - The effects of an H3 agonist, R-alpha-methylhistamine (alpha-MeHA), and an H3 antagonist, thioperamide, on monoamine oxidase (MAO) activity in the hypothalamus of rat, monkey and human brains were compared in vitro. The histamine H(3) receptor ligands competitively inhibited MAO-B, but noncompetitively inhibited MAO-A in all three mammalian species. However, alpha-MeHA inhibited MAO-A more potently than MAO-B at high concentrations in all three species. The K(i) values for MAO-A of alpha-MeHA in hypothalamic homogenates of rat, monkey and human brains were estimated to be 1.1, 1.2 and 1.9 mM, respectively, suggesting that alpha-MeHA cannot behave as a substrate for the MAO inhibitor. In contrast, rat, monkey and human brain MAO-B activities were inhibited by thioperamide, with respective K(i) values of 174.6, 8.2 and 10.8 microM, more potently than MAO-A activity. These results indicate that thioperamide, which elicits a strong activation of histamine release and turnover to N-tele-methylhistamine from histamine, competitively inhibits the conversion of N-tele-methylhistamine to N tele-methylimidazoleacetic acid in human and monkey brains where MAO-B predominates. PMID- 11430878 TI - FK506 ameliorates the discrimination learning impairment due to preventing the rarefaction of white matter induced by chronic cerebral hypoperfusion in rats. AB - We examined the effects of the immunosuppressant tacrolimus (FK506) on the discrimination learning impairment induced by chronic cerebral hypoperfusion in rats. Chronic cerebral hypoperfusion was prepared by permanent ligation of bilateral common carotid arteries for male Wistar rats aged 9 weeks. FK506 (0.05 mg/kg, s.c.) recovered the learning impairment and also prevented the rarefaction of white matter and striatal neuronal cell damage. Our findings suggest that FK506 ameliorates the learning impairment mainly due to preventing neuropathological alterations. PMID- 11430879 TI - Responses of ventral tegmental area GABA neurons to brain stimulation reward. AB - Dopamine neurons in the ventral tegmental area (VTA) have been implicated in rewarded behaviors, including intracranial self-stimulation (ICSS). We demonstrate, in unrestrained rats, that the discharge activity of a homogeneous population of presumed VTA GABA neurons, implicated in cortical arousal, increases before ICSS of the medial forebrain bundle (MFB). These findings suggest that VTA GABA neurons may be involved in the attentive processes related to brain stimulation reward (BSR). PMID- 11430880 TI - A possible role for BDNF, NT-4 and TrkB in the spinal cord and muscle of rat subjected to mechanical overload, bupivacaine injection and axotomy. AB - Neurotrophins play a crucial role in the regulation of survival and the maintenance of specific functions for various populations of neurons. Neurotrophin-4 (NT-4) is most abundant in skeletal muscle, and is thought to promote sciatic nerve sprouting, inhibit agrin-induced acetylcholine receptor (AChR) clustering, evoke postsynaptic potentiation and induce mitochondrial proliferation. Using Western blot analysis, immunoprecipitation and immunohistochemistry, we investigated the distribution of NT-4 in slow- and fast type muscles. We also tested the adaptive response of this protein in the mechanically overloaded muscle, in the regenerating muscle following bupivacaine injection and in the denervated muscle. Additionally, we investigated whether TrkB phosphorylation in the spinal cord and in the sciatic nerve occurs through the interaction with BDNF or NT-4 when the innervating muscle is damaged. Markedly more NT-4 was expressed in fast-type muscles compared with the slow types. TrkB protein was more frequently observed around the edge of myofibers (neuromuscular junction) of the soleus muscle compared with the gastrocnemius muscle. TrkB tyrosine phosphorylation occurred in the spinal cord but not in the sciatic nerve 24 h after bupivacaine injection of the innervating muscle. At the same time, the amount of TrkB co-precipitating with BDNF was markedly increased in the spinal cord. A rapid activation of TrkB (1-8 h) was also observed in the spinal cord after axotomy,while the amount of TrkB co-precipitating with NT-4 was markedly lower after axotomy. These results indicate that NT-4 is preferentially distributed in fast-type muscles. Furthermore, by interacting with BDNF and NT-4, the TrkB in the spinal cord may be important for the survival of motoneurons and outgrowth of injured peripheral axons following muscle damage. PMID- 11430881 TI - Anesthesia alters NO-mediated functional hyperemia. AB - Many properties of nitric oxide, NO, (localization, diffusiveness, half-life, vasodilatory affects) have supported its potential role in mediating the link between local cerebral activity and blood flow. However, evidence that both supports and refutes a role for NO in functional hyperemia have been presented. The present study employed multiple nitric oxide synthase inhibitors, two anesthetic regimes and laser-Doppler flowmetry to test the hypothesis that NO is critically involved in mediating the functional hyperemic response within rodent whisker-barrel cortex (WBC). In urethane anesthetized animals, functional hyperemic responses were obtained both before and after 1 mg/kg atropine infusion, 30 mg/kg i.v. L-NAME (N-Nitro-L-arginine methylester) infusion, 30 mg/kg L-NA (N-Nitro-L-arginine) infusion or 25 mg/kg 7-NI (7-nitroindazole). L NAME was also tested in a group of animals pretreated with halothane before urethane anesthesia. Neither the magnitude of the blood flow response nor its time course was altered by NO blockade or atropine administration when compared to pre-infusion controls in urethane anesthetized rats. In contrast, animals that were pretreated with halothane exhibited a 33% inhibition of functional hyperemia after L-NAME administration. Taken together, these data do not support a primary role for NO in rat WBC functional hyperemia and suggest that previous reports of inhibition may have been secondary to the anesthesia employed. PMID- 11430882 TI - Orexins: effects on behavior and localisation of orexin receptor 2 messenger ribonucleic acid in the rat brainstem. AB - The orexins are neuropeptides originally reported to be involved in the stimulation of food intake. However, analysis of orexin immunoreactive fibres have revealed the densest innervation in brain sites involved in arousal and sleep-wake control, notably the noradrenergic locus coeruleus, an area that also expresses orexin receptor 1 (OX1R) messenger RNA (mRNA). We report here that, in the rat, a single intracerebroventricular injection of orexin A (1 and 3 nmol) or orexin B (3 nmol), during the early light phase, did not increase food intake over the first 4 h postinjection. However, the frequency of active behaviors such as grooming, rearing, burrowing and locomotion increased. Feeding behavior and food intake subsequently decreased over the following 20 h (4-24 h postinjection period) in the orexin A 3 nmol injected group whilst the frequency of inactive behavior (still or asleep) in this group increased. Using riboprobes, we performed in situ hybridization histochemistry to map the distribution of orexin receptor 2 (OX2R) mRNA within the rat brainstem. We report here, for the first time, the presence of OX2R mRNA in the nucleus of the solitary tract and the lateral reticular field (LRt). The LRt is a brainstem site that, amongst other functions, is implicated in attention and wakefulness. This distribution of OX2R and the effects on behavior support recent reports that the orexins might modulate central nervous system arousal and sleep-wake mechanisms rather than exclusively being involved in the control of food intake. PMID- 11430883 TI - Hyperactivity, neuromotor defects, and impaired learning and memory in a mouse model for metachromatic leukodystrophy. AB - Deficiency of arylsulfatase A (ASA) causes the autosomal recessive lipidosis, metachromatic leukodystrophy (MLD). Performance on tests of activity, motor ability and learning/memory was assessed in ASA-deficient mice and normal controls at 3, 6 and 12 months-of-age. ASA-deficient mice showed consistently increased cage activity in all age groups, whereas open field activity was increased only in the 3-month-old group. Motor coordination and equilibrium, as tested in the rotarod test, was impaired in 12-month-old ASA-deficient mice. Passive avoidance learning was tested in the step-through box. Performance on this test was impaired in the 12-month-old group only. Spatial learning and memory abilities were tested in the Morris water maze. Six-month-old ASA deficient mice displayed slightly impaired hidden-platform acquisition performance. Three-month-old animals, on the other hand, did not show any acquisition or retention defect on this task, notwithstanding significantly reduced swimming velocity. Acquisition training, both in the hidden- and visible platform conditions of the Morris water maze, and retention performance during the probe trials were impaired in 12-month-old ASA-deficient mice. The hyperactivity, motor incoordination and slowing, and the age-related learning/memory defects, reported here in ASA-deficient mice, may relate to the decline of neuromotor and cognitive functions in MLD patients, and could be used as correlative or outcome measures in the study of MLD pathophysiology and treatment. PMID- 11430884 TI - Deposition of Alzheimer's vascular amyloid-beta is associated with decreased expression of brain L-3-hydroxyacyl-coenzyme A dehydrogenase (ERAB). AB - L-3-hydroxyacyl-coenzyme A dehydrogenase type II (HADH) was described as an endoplasmic reticulum amyloid beta-peptide-binding protein (ERAB), which enhances Abeta toxicity, and accumulates in neurons in Alzheimer's disease (AD). Hence, HADH/ERAB was suggested to mediate the amyloid-induced neurodegeneration. We estimated the in vivo interactions of HADH and Abeta in an immunocytochemical study of ten Alzheimer's disease and seven normal brains using five monoclonal HADH-specific antibodies. We found no HADH in amyloid plaques or vascular amyloid. The neuronal expression of HADH was not correlated with the severity of amyloid load in neuropil. HADH was expressed in vascular smooth muscle cells in young and old controls and in amyloid-free blood vessels in AD cases, but little or no HADH was in smooth muscle cells in arteries with amyloid deposits. The putative intracellular interaction between HADH and Abeta in amyloid-producing cells was further studied in vascular smooth muscle cells isolated from brain blood vessels with amyloid-beta angiopathy - the cells that were shown previously to accumulate Abeta intracellularly ['Research advances in Alzheimer's disease and related disorders' (1995) 747; Brain Res. 676 (1995) 225; Neurosci. Lett. 183 (1995) 120]. HADH had a mitochondrial localization and did not co-localize with an endoplasmic reticulum marker. Cells that accumulated Abeta were those with low expression of HADH and the proteins did not co-localize. Explanation of the association between low levels of HADH and deposition of Abeta by brain smooth muscle cells requires further studies. PMID- 11430885 TI - Regional and age-dependent expression of the nitric oxide receptor, soluble guanylyl cyclase, in the human brain. AB - Nitric oxide (NO), synthesized by neuronal NO synthase (NOS-I), plays essential physiological roles in the brain. The major molecular target for NO is soluble guanylyl cyclase (sGC), a heterodimeric hemoprotein composed of a larger alpha and a smaller beta subunit. Both subunits of sGC are needed to generate the second messenger cyclic GMP (cGMP). Here we show using subunit-specific antibodies and Western blot analysis that sGCalpha1 and sGCbeta1 protein subunits are present in all examined human brain regions. The relative distribution of the two subunits was similar and also correlated well with the known distribution of NOS-I. The highest expression levels of sGC were found in cortex, basal ganglia and the limbic system. These regions display the most prominent biochemical and histological changes during ageing. In cortex, a negative correlation between the amounts of sGC and age was found, while sex and post-mortem delay time did not affect sGC levels significantly. Our data suggest that sGCalpha1 and sGCbeta1 subunits are widely distributed in human brain, consistent with a major role in NO signaling. Moreover, the NO/cGMP pathway appears to be affected by ageing in the human brain. PMID- 11430886 TI - Autoradiographic localization of N-type VGCCs in gerbil hippocampus and failure of omega-conotoxin MVIIA to attenuate neuronal injury after transient cerebral ischemia. AB - In the mammalian central nervous system, transient global ischemia of specific duration causes selective degeneration of CA1 pyramidal neurons in hippocampus. Many of the ischemia-induced pathophysiologic cascades that destroy the neurons are triggered by pre- and postsynaptic calcium entry. Consistent with this, many calcium channel blockers have been shown to be neuroprotective in global models of ischemia. omega-Conotoxin MVIIA, a selective N-type VGCC blocker isolated from the venom of Conus magus, protects CA1 neurons in the rat model of global ischemia, albeit transiently. The mechanism by which this peptide renders neuroprotection is unknown. We performed high-resolution receptor autoradiography with the radiolabeled peptide and observed highest binding in stratum lucidum of CA3 subfield, known to contain inhibitory neurons potentially important in the pathogenesis of delayed neuronal death. This finding suggested that the survival of stratum lucidum inhibitory neurons might be the primary event, leading to CA1 neuroprotection after ischemia. Testing of this hypothesis required the reproduction of its neuroprotective effects in the gerbil model of global ischemia. Surprisingly, we found that omega-MVIIA did not attenuate CA1 hippocampal injury after 5 min of cerebral ischemia in gerbil. Possible reasons are discussed. Lastly, we show that the peptide can be used as a synaptic marker in assessing short and long-term changes that occur in hippocampus after ischemic injury. PMID- 11430887 TI - Decline in motor functions in aging is related to the loss of NMDA receptors. AB - The aim of the study was to assess the contribution of central dopaminergic and glutamatergic systems to the age-dependent loss of motor functions in rats. Rats of three age groups were compared: young (3-5-month-old), middle-aged (20-21 month-old) and old (29-31-month-old). The obtained results showed an age dependent decline in the electromyographic (EMG) resting and reflex activities in the gastrocnemius and tibialis anterior muscles, as well as in the T-maze performance. Although these disturbances were accompanied with significant age dependent decreases in the binding to NMDA, AMPA and dopamine D2 receptors, and a decline in the number of nigral dopamine neurons, they were significantly correlated with the loss of the binding to NMDA receptors only. The reduction in T-maze performance with aging was additionally correlated with a decrease in motor functions (EMG activity). The study suggests a crucial role of the loss of NMDA receptors in age-dependent motor disabilities, as well as in disturbances measured in the T-maze. PMID- 11430888 TI - Effects of social stimuli on sleep in mice: non-rapid-eye-movement (NREM) sleep is promoted by aggressive interaction but not by sexual interaction. AB - Sleep is generally considered to be a process of recovery from prior wakefulness. In addition to being affected by the duration of the waking period, sleep architecture and sleep EEG also depend on the quality of wakefulness. In the present experiment, we examined how sleep is affected by different social stimuli (social conflict and sexual interaction). Male C57BL/6J mice were placed in the cage of an aggressive dominant male or an estrous female for 1 h in the middle of the light phase. The conflict with an aggressive male had a pronounced NREM sleep promoting effect. EEG slow wave activity, a measure of NREM sleep intensity, was increased for about 6 h and NREM sleep time was significantly increased for 12 h. REM sleep was strongly suppressed during the remainder of the light phase after the conflict, followed by a rebound later in the recovery phase. The sexual interaction, in contrast, had only mild effects. Both NREM sleep and REM sleep were somewhat suppressed shortly after the interaction. In a separate group of mice, blood samples were taken to measure prolactin and corticosterone. The results suggest that the temporary suppression of REM sleep following the social stimuli may be partly due to elevated corticosterone. The different effects of the social stimuli on NREM sleep are not easily explained by differences in the hormone responses. In conclusion, although both social conflict and sexual interaction induce a strong physiological activation, only social conflict has a strong stimulatory effect on NREM sleep mechanisms. PMID- 11430889 TI - Adenosine activates ATP-sensitive K(+) currents in pericytes of rat retinal microvessels: role of A1 and A2a receptors. AB - In the CNS, contractile pericytes are positioned on the endothelial walls of microvessels where they are thought to play a role in adjusting blood flow to meet local metabolic needs. This function may be particularly important in the retina where pericytes are more numerous than at any other site. Despite the putative importance of pericytes, knowledge of the mechanisms by which vasoactive molecules, such as adenosine, regulate their function is limited. Using the perforated-patch configuration of the patch-clamp technique to monitor the whole cell currents of pericytes located on microvessels freshly isolated from the adult rat retina, we found that adenosine reversibly activated a hyperpolarizing current in 98% of the sampled pericytes. This adenosine-induced current is likely to be due to the opening of ATP-sensitive potassium (K(ATP)) channels since it had a reversal potential near the equilibrium potential for K(+), was inhibited by the K(ATP) channel blocker, glibenclamide, and was mimicked by pinacidil, which is a K(ATP) channel opener. Experiments with specific agonists and antagonists indicated that both the high affinity A1 and the lower affinity A2a adenosine receptors provided effective pathways for activating K(ATP) currents in pericytes recorded under normal metabolic conditions. However, during chemical ischemia, the A1 receptor pathway rapidly became ineffective. In contrast, activation of A2a adenosine receptors continued to open K(ATP) channels in ischemic pericytes. These results suggest that the regulation of K(ATP) channels via A1 and A2a receptors allows adenosine to serve over a broad range of metabolic conditions as a vasoactive signal in the retinal microvasculature. PMID- 11430890 TI - Serotonergic lesion of median raphe nucleus alters nerve growth factor content and vulnerability of cholinergic septohippocampal neurons in rat. AB - About 45% of the serotonergic raphe neurons are reported to express nerve growth factor (NGF) receptors. We therefore investigated whether selective serotonergic lesions of the median or dorsal raphe nuclei are associated with changes in NGF protein levels of the brain and whether the loss of serotonergic function alters the vulnerability of cholinergic septohippocampal neurons. In adult rats the hippocampal NGF content changed in a biphasic way after lesion of the median raphe nucleus by 5,7-dihydroxytryptamine (5,7-DHT), with a significant increase after 2-3 weeks of up to 35%, followed by a significant reduction of 22% below control levels after 7 weeks, and a return to control levels within the following 4 weeks. By contrast, the decrease in hippocampal serotonin and 5 hydroxyindoleacetic acid remained throughout the observation period of 11 weeks, being still reduced to 15 and 30% of the control levels, respectively. In the frontal cortex the partial loss of the serotonergic innervation projecting from the median raphe was associated 5 weeks after 5,7-DHT injection with an increase in NGF protein of 39.7+/-9.6% (P<0.05), which remained elevated up to 11 weeks. At 9 weeks after 5,7-DHT, the lesion of the septohippocampal cholinergic neurons induced by the cholinotoxin ethylcholine aziridinium (AF64A) was exaggerated (P<0.05) as compared to AF64A-treated rats with intact serotonergic innervation. The present data indicate that a serotonergic lesion of the median raphe nucleus results in biphasic changes of NGF protein content and in a delayed increase in the vulnerability of septohippocampal cholinergic neurons. PMID- 11430891 TI - Analgesia elicited by OFQ/nociceptin and its fragments from the amygdala in rats. AB - The heptadecapeptide, orphanin FQ/nociceptin (OFQ/N), binds with high affinity to the ORL-1/KOR-3 opioid receptor clone, yet binds poorly with traditional opioid receptors. OFQ/N has a complex functional profile with relation to nociceptive processing, displaying pro-nociceptive properties in some studies, acting as an inhibitor of stress-induced analgesia in others, yet producing both spinal and supraspinal antinociceptive actions in other studies. Among the intracerebral sites at which OFQ/N might produce one or more of these actions is the amygdala which has been intimately implicated in both antinociceptive and stress-related responses. Therefore, the present study assessed whether microinjections into the amygdala of equimolar doses of OFQ/N(1-17) or its shorter-chained active fragments, OFQ/N(1-11) or OFQ/N(1-7), would produce analgesia as measured by either reactivity to high-intensity radiant heat or reactivity to electric shock, and produce hyperalgesia as measured by reactivity to lower-intensity radiant heat. OFQ/N(1-17) in the amygdala produced a dose-dependent and time-dependent increase in high-intensity tail-flick latencies with maximal effects observed at a dose range of 0.75-3 nmol, and lesser effects at lower (0.015-0.15 nmol) and higher (5.5-30 nmol) doses. Both OFQ/N(1-11) and OFQ/N(1-7) in the amygdala displayed lower magnitudes of analgesia than OFQ/N(1-17) on this measure, with OFQ/N(1-11) displaying maximal effects at higher (15-30 nmol) doses and OFQ/N(1 7) displaying maximal effects at lower (0.15-1.5 nmol) doses. In contrast to traditional mu and kappa opioids and beta-endorphin, none of the OFQ/N fragments in the amygdala exhibited any analgesic responses on the jump test. Finally, using a low-intensity radiant heat assay capable of detecting hyperalgesic responses, each of the OFQ/N fragments in the amygdala increased tail-flick latencies on this measure. Therefore, OFQ/N fragments appear to exert only analgesic responses in the amygdala with quantitative and qualitative differences relative to traditional opioid agonists. PMID- 11430892 TI - The NMDA receptor antagonist MK-801 reduces Fos-like immunoreactivity within the trigeminocervical complex following superior sagittal sinus stimulation in the cat. AB - Expression of Fos protein is an indicator of neuronal perturbation and is readily observed in the caudal medulla and the spinal cord following trigeminovascular nociceptive activation by electrical stimulation of the superior sagittal sinus (SSS) in the cat. It has been shown in the rat that N-methyl-D-aspartate (NMDA) receptor blockade causes a reduction in Fos protein expression after generalised meningeal irritation. We wished to examine if the same relationship was true in the cat, using the same non-competitive NMDA receptor antagonist MK-801, and a trigeminovascular-specific stimulus. A group of experimental animals underwent stimulation following blinded administration of MK-801 (4 mg/kg i.v.); control animals underwent stimulation minus MK-801, and a non-stimulated control animal underwent surgery alone. The regions examined for Fos-like immunoreactivity were the trigeminal nucleus caudalis (TNC) and its caudal extension into the C(1) and C(2) levels of the upper cervical spinal cord. The Fos-positive cell counts for the three regions (TNC, C(1) and C(2)) were grouped together for analysis. In the control stimulated group a median of 78 (56-99, quartile range, n=4) cells were Fos-positive. In the group treated with MK-801 the median number of Fos-positive cells was reduced to 40 (30-48; P<0.03, n=7). The large reduction that was observed in SSS stimulation-evoked Fos protein expression following the administration of MK-801, taken together with electrophysiological data, indicates a role for glutamate in neurotransmission within the trigeminocervical complex. Understanding glutamatergic mechanisms in the trigeminocervical complex offers mechanistic insight and therapeutic possibilities for primary neurovascular headaches, such as migraine. PMID- 11430893 TI - Identification of central sites involved in butorphanol-induced feeding in rats. AB - Butorphanol (BT), a mixed kappa- and mu-opioid receptor agonist, induces vigorous food intake in rats. Peripheral injection of BT seems to increase food intake more effectively than intracerebroventricular administration. To further elucidate the nature of BT's influence on consummatory behavior, we examined which feeding-related brain areas exhibit increased c-Fos immunoreactivity (IR) following subcutaneous injection of 4 mg/kg body weight BT, a dose known to induce a maximal orexigenic response. We also evaluated whether direct administration of BT into the forebrain regions activated by peripheral BT injection affects food intake. Peripheral BT administration induced c-Fos-IR in the hypothalamic paraventricular nucleus (PVN), central nucleus of the amygdala (CeA), and nucleus of the solitary tract (NTS). However, 0.1-30 microg BT infused into the CeA, failed to increase food intake 1, 2, and 4 h after injection. Only the highest dose of BT (30 microg) injected into the PVN increased feeding. These results suggest that the PVN, CeA, and NTS mediate the effects of peripherally injected BT. The PVN or CeA are probably not the main target sites of immediate BT action. PMID- 11430894 TI - Expression of polysialylated neural cell adhesion molecule in rat brain after transient middle cerebral artery occlusion. AB - The highly polysialylated form of neural cell adhesion molecule (PSA-NCAM) is important for neurite outgrowth. With this molecule as a marker of plastic change in neurons, we investigated its temporal expression in rat brain after transient middle cerebral artery (MCA) occlusion. In sham-control brain, only subependymal neurons showed a positive immunoreactivity for PSA-NCAM. After 90 min of transient MCA occlusion, neurons in the piriform cortex began to be positively stained at 1 h, while neurons in the cortex and caudate of the MCA territory became positive after 8 h. The stainings persisted for 1 and 3 days after reperfusion. The present results indicate that neurons in the cerebral cortex and caudate have the capability of plastic change in the adult brain, and that those in the piriform cortex rapidly undergo plastic change probably in response to transneuronal injury. PMID- 11430895 TI - Biochemical and pathological study of endogenous 1-benzyl-1,2,3,4 tetrahydroisoquinoline-induced parkinsonism in the mouse. AB - We administered 1-benzyl-1,2,3,4-tetrahydroisoquinoline (1-BnTIQ; 80 mg/kg, i.p.), an endogenous neurotoxin known to cause bradykinesia, the Parkinson's disease-like symptom, in order to obtain biochemical and pathological evidence of behavioral abnormalities. Immunohistochemical analysis demonstrated that 1-BnTIQ did not decrease the number of tyrosine hydroxylase-positive cells in the substantia nigra. Biochemical analysis demonstrated significantly increased striatal dopamine (DA) content, while DA metabolites in the striatum remained at control levels. We concluded that the 1-BnTIQ-induced bradykinesia has a different mechanism of action than that underlying the MPTP-induced depletion of striatal DA neurons. PMID- 11430896 TI - Gap junctions synchronize the firing of inhibitory interneurons in guinea pig hippocampus. AB - The convulsant 4-aminopyridine (4AP) facilitates the synchronous firing of interneurons in the hippocampus, eliciting giant inhibitory postsynaptic potentials (IPSPs) in CA3 pyramidal cells. We used the gap junction blocker carbenoxolone to investigate the role of electrotonic coupling in both the initiation and the maintenance of 4AP-facilitated inhibitory circuit oscillations. Carbenoxolone abolished all synchronized IPSPs in CA3 cells elicited by 4AP in the presence of ionotropic glutamate receptor blockers. Carbenoxolone also blocked the isolated synchronized GABA(B) IPSPs generated in CA3 cells by a subpopulation of interneurons. These data confirm that: (1) the interneurons producing GABA(B) responses in CA3 cells are electrotonically coupled, and (2) gap junctions among interneurons are essential for initiating synchronized interneuron oscillatory firing in 4AP. PMID- 11430897 TI - Long-term potentiation protects rat hippocampal slices from the effects of acute hypoxia. AB - We have previously shown that long-term potentiation (LTP) decreases the sensitivity of glutamate receptors in the rat hippocampal CA1 region to exogenously applied glutamate agonists. Since the pathophysiology of hypoxia/ischemia involves increased concentration of endogenous glutamate, we tested the hypothesis that LTP could reduce the effects of hypoxia in the hippocampal slice. The effects of LTP on hypoxia were measured by the changes in population spike potentials (PS) or field excitatory post-synaptic potentials (fepsps). Hypoxia was induced by perfusing the slice with (i) artificial CSF which had been pre-gassed with 95%N2/5% CO2; (ii) artificial CSF which had not been pre-gassed with 95% O2/5% CO2; or (iii) an oxygen-glucose deprived (OGD) medium which was similar to (ii) and in which the glucose had been replaced with sucrose. Exposure of a slice to a hypoxic medium for 1.5-3.0 min led to a decrease in the PS or fepsps; the potentials recovered to control levels within 3 5 min. Repeat exposure, 45 min later, of the same slice to the same hypoxic medium for the same duration as the first exposure caused a reduction in the potentials again; there were no significant differences between the degree of reduction caused by the first or second exposure for all three types of hypoxic media (P>0.05; paired t-test). In some of the slices, two episodes of LTP were induced 25 and 35 min after the first hypoxic exposure; this caused inhibition of reduction in potentials caused by the second hypoxic insult which was given at 45 min after the first; the differences in reduction in potentials were highly significant for all the hypoxic media used (P<0.01; paired t-test). The neuroprotective effects of LTP were not prevented by cyclothiazide or inhibitors of NO synthetase compounds that have been shown to be effective in blocking the effects of LTP on the actions of exogenously applied AMPA and NMDA, respectively. The neuroprotective effects of LTP were similar to those of propentofylline, a known neuroprotective compound. We conclude that LTP causes an appreciable protection of hippocampal slices to various models of acute hypoxia. This phenomenon does not appear to involve desensitisation of AMPA receptors or mediation by NO, but may account for the recognised inverse relationship between educational attainment and the development of dementia. PMID- 11430898 TI - DOI, a 5-HT2A/2C receptor agonist, attenuates clozapine-induced cortical dopamine release. AB - (+/-)-1-(2,5-Dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride (DOI, 1.25, 2.5 and 5 mg/kg), a serotonin (5-HT)2A/2C agonist, produced an inverted U-shaped increase in DA release in rat medial prefrontal cortex (mPFC) with a significant effect only at 2.5 mg/kg. This effect was completely abolished by M100907 (0.1 mg/kg), a 5-HT2A antagonist, and WAY100635 (0.2 mg/kg), a 5-HT1A antagonist, neither of which when given alone affected dopamine release. DOI (2.5 mg/kg), but not the 5-HT2C agonist Ro 60-0175 (3 mg/kg), attenuated clozapine (20 mg/kg) induced mPFC dopamine release. These results suggest that 5-HT2A receptor stimulation increases basal cortical dopamine release via 5-HT1A receptor stimulation, and inhibits clozapine-induced cortical dopamine release by diminishing 5-HT2A receptor blockade. PMID- 11430899 TI - Selective activation of accumbens neurons by cocaine-associated stimuli during a water/cocaine multiple schedule. AB - Electrophysiological recording procedures were used to examine the responsiveness of nucleus accumbens (Acb) neurons to stimuli associated with cocaine delivery during a multiple schedule for water reinforcement and cocaine self administration. Rats (n=9) were trained to press one lever for water reinforcement (0.05 ml/resp.; fixed ratio 1; FR1; 15 min) and a second spatially distinct lever for intravenous cocaine (0.33 mg/infusion; FR1; 2 h). The delivery of each reinforcer was signaled by different auditory stimuli. Of 101 neurons, 52 cells were classified as phasically active, exhibiting one of four well-defined types of patterned discharges relative to the water- or cocaine-reinforced response [J. Neurosci., 14(12) (1994) 7735; J. Neurosci., 20(11) (2000) 4255]. Acb cells were examined in test sessions consisting of 'probe' trials during which the stimulus previously paired with cocaine infusion was randomly presented in a response-independent manner during the self-administration portion of the session. Results showed that only neurons that exhibited changes in firing rate within seconds following the reinforced response for cocaine (but not water) were activated by the stimulus. This finding indicates that the responsiveness of cocaine selective neurons to cocaine-associated stimuli likely represents a conditioned response as opposed to a generalized stimulus-evoked discharge. PMID- 11430900 TI - Differential expression of CPD1 during postnatal development in the mouse cerebellum. AB - Several regulated mRNAs were detected by applying differential display to the mouse cerebellum during postnatal development. One cDNA fragment, referred to as CPD1 (GenBank U89345), was characterized and cloned. Northern blots showed maximum mRNA expression at postnatal day seven (P7). The mRNA encodes a protein of 260 amino acids. In situ RT-PCR showed that CPD1 is expressed mainly in granule cells and faintly in Purkinje cells. Polyclonal rabbit antibodies and oligobodies (oligonucleotide-based synthetic antibodies) revealed a protein of 34 kDa in Western blots. Immunohistochemistry showed not only marked nuclear staining but also mild cytoplasmic localization. Granule cells undergoing active division (P4) showed very little expression of CPD1 protein, which increases from P7 to P17. CPD1, affinity-purified using a chemically synthesized oligobody inhibits the activity of protein phosphatase PP2A but not protein phosphatase PP1. Differentiated PC12 cells also showed nuclear and cytoplasmic localization. Interestingly, maximal cytoplasmic CPD1/PP2A colocalization was observed near cell membrane regions that are far from growing neurites, and on growing cones. These results suggest that CPD1 might have an important role in cerebellar development. PMID- 11430901 TI - Quantitative analysis of cell death and ferritin expression in response to cortical iron: implications for hypoxia-ischemia and stroke. AB - Iron that is not bound to storage proteins can catalyse the generation of toxic hydroxyl radicals. Iron can be released from brain storage proteins by hypoxic conditions, such as those that accompany stroke, and the situation can be compounded by iron released from hemoglobin in extravasated blood cells. Despite the neurotoxicity of iron, there is little quantitative data concerning the spatio-temporal extent of its toxicity in vivo. The present study measures the effects of a pathologically relevant concentration of iron (1.0 mM) on neuronal death and on ferritin expression in vivo. Injection of iron (1 microl ferric ammonium citrate) into rat parietal cortex resulted in 7.9-fold more ferritin labeled cells than did control injections of ammonium citrate at 1 day post injection. This elevated expression continued for at least 1 week. One day after injection, the mean number of Fluoro-Jade-labeled degenerating neurons in 100 microm sections passing through the center of ferric ammonium citrate injection sites was 664+/-64. This value was 4.5-fold higher than at ammonium citrate injection sites, and this difference increased to 56-fold by day three. By 5 days post-injection, few dying neurons were observed at the control sites, but neurodegeneration continued beyond a week at the iron-injected sites. Thus, iron released during a brief episode of hypoxia-ischemia or during a stroke may be neurotoxic for a protracted period. Therefore, our findings indicate that it may be beneficial to target iron-induced peroxidation throughout the first few weeks following an intracerebral hemorrhage or an hypoxic-ischemic episode. PMID- 11430902 TI - Neuroprotective effects of extracellular glutamate are absent in hippocampal organotypic cultures treated with the amyloid peptide Abeta(25-35). AB - Hippocampal cells are particularly vulnerable in Alzheimer's disease but the cause of cell death is unknown. Amyloid toxicity has been implicated in hippocampal cell death, but its specific mechanisms are poorly understood. We used confocal microscopy to examine the effects of the amyloid peptide fragment 25-35 (Abeta(25-35)) on cell death in organotypic hippocampal slice cultures. Addition of glutamate to the culture medium significantly improved nerve cell survival in cultures subjected to consecutive medium exchanges. This effect was lost if cultures were treated with the amyloid peptide fragment Abeta(25-35) but not the inactive peptide 35-25. These data suggest that one of the mechanisms responsible for amyloid toxicity may be inhibition of the survival promoting effects of extracellular glutamate. PMID- 11430903 TI - Serotonin projection patterns to the cochlear nucleus. AB - The cochlear nucleus is well known as an obligatory relay center for primary auditory nerve fibers. Perhaps not so well known is the neural input to the cochlear nucleus from cells containing serotonin that reside near the midline in the midbrain raphe region. Although the specific locations of the main, if not sole, sources of serotonin within the dorsal cochlear nucleus subdivision are known to be the dorsal and median raphe nuclei, sources of serotonin located within other cochlear nucleus subdivisions are not currently known. Anterograde tract tracing was used to label fibers originating from the dorsal and median raphe nuclei while fluorescence immunohistochemistry was used to simultaneously label specific serotonin fibers in cat. Biotinylated dextran amine was injected into the dorsal and median raphe nuclei and was visualized with Texas Red, while serotonin was visualized with fluorescein. Thus, double-labeled fibers were unequivocally identified as serotoninergic and originating from one of the labeled neurons within the dorsal and median raphe nuclei. Double-labeled fiber segments, typically of fine caliber with oval varicosities, were observed in many areas of the cochlear nucleus. They were found in the molecular layer of the dorsal cochlear nucleus, in the small cell cap region, and in the granule cell and external regions of the cochlear nuclei, bilaterally, of all cats. However, the density of these double-labeled fiber segments varied considerably depending upon the exact region in which they were found. Fiber segments were most dense in the dorsal cochlear nucleus (especially in the molecular layer) and the large spherical cell area of the anteroventral cochlear nucleus; they were moderately dense in the small cell cap region; and fiber segments were least dense in the octopus and multipolar cell regions of the posteroventral cochlear nucleus. Because of the presence of labeled fiber segments in subdivisions of the cochlear nucleus other than the dorsal cochlear nucleus, we concluded that the serotoninergic projection pattern to the cochlear nucleus is divergent and non specific. Double-labeled fiber segments were also present, but sparse, in the superior olive, localized mainly in periolivary regions; this indicated that the divergence of dorsal and median raphe neurons that extends throughout regions of the cochlear nucleus also extended well beyond the cochlear nucleus to include at least the superior olivary complex as well. PMID- 11430904 TI - 1H magnetic resonance spectroscopic imaging of permanent focal cerebral ischemia in rat: longitudinal metabolic changes in ischemic core and rim. AB - The purpose of this study was to determine whether regional differences in metabolites can be seen chronologically in permanent focal cerebral ischemia using 1H magnetic resonance spectroscopic imaging (MRSI), and whether these changes reflect pathological outcome. Regional variation in metabolites after permanent focal ischemia were investigated longitudinally in rats using 1H MRSI for a total of 7 days and then compared to histopathological findings. Four hours after the induction of ischemia, N-acetyl-L-aspartate (NAA) levels in the lateral caudo-putamen and the somatosensory cortex, core ischemic regions, decreased 22 and 40%, respectively. This reduction in NAA was coupled with a marked rise in lactate. In the medial caudo-putamen, the ischemic rim, however, NAA was preserved in spite of a marked increase in lactate. By 24 h post ischemia, the levels of NAA in medial caudo-putamen (ischemic rim in caudate) also decreased significantly. However NAA in cingulated cortex (ischemic rim in cortex) decreased more gradually between 24 and 48 h. This regional difference can reflect the severity of metabolic derangement in the acute stage. After 96 h following ischemia, the levels of all metabolites detected by 1H MRSI had decreased and the levels of NAA decline reflected the severity of histopathological damage. In conclusion, the regional metabolic differences could be assessed by 1H MRSI chronologically, and the depth of NAA decline reflected histopathological changes in the chronic stage. PMID- 11430905 TI - Expression of amyloid precursor protein, tau and presenilin RNAs in rat hippocampus following deafferentation lesions. AB - In this study, entorhinal cortex lesions and/or medial septal area cholinergic lesions were used in the rat to mimic some of the principal and earliest affects in Alzheimer's disease, namely hippocampal deafferentation. We wished to test the hypothesis that deafferentation lesions cause changes in the regulation of three proteins that are known to be important in Alzheimer's disease pathology, namely amyloid precursor protein, presenilin and tau. Expression of amyloid precursor protein mRNA was increased in several subfields of hippocampus when examined 1 week after entorhinal cortex lesion, but was reduced, compared to sham operated controls, after medial septal area cholinergic lesions. Cholinergic lesions were combined with entorhinal cortex lesions and produced no change in APP mRNA levels compared to controls. No significant changes were observed in the parietal cortex after entorhinal cortex or cholinergic lesions either alone or in combination. Tau mRNA level in hippocampus was unchanged after lesions. Presenilin-1 mRNA was expressed in the hippocampus at very low levels, and appeared to be increased following entorhinal cortex lesion. Our results support the hypothesis that amyloid precursor protein expression in hippocampal neurons is differentially affected by glutamatergic and cholinergic afferent input, and that presenilin-1, but not tau, may be subject to the same type of control in vivo. PMID- 11430906 TI - Genetic manipulation of key determinants of ion flow in glutamate receptor channels in the mouse. AB - Glutamate receptor channels are built around an ancient pore loop structure which defines the inner channel environment and which is connected to structures for channel gating. This pore loop, which corresponds to the M2 region of the receptor subunits, enters the lipid bilayer from the intracellular side in an alpha-helical configuration, then kinks to form a random coil and exits the lipid bilayer at the intracellular side. The narrow constriction of the channel is formed by amino acid residues that occupy a position shortly after the end of the alpha-helical part of M2. These residues determine ion selectivity and conductance properties of the glutamate-gated channel. The critical residues are asparagines for NMDA receptor subunits and glutamine or arginine for AMPA and kainate receptor subunits. Presence of arginine in the critical channel position of AMPA and kainate receptors is controlled by site-selective RNA editing. To study the importance of these critical channel residues in the mouse, we introduced codon changes in the endogenous genes for NMDA and AMPA receptor subunits. Our results show that changes in the critical channel position are not tolerated, but lead to early death. Therefore, the impact on adult synaptic function and plasticity by glutamate receptor channels with changed ion selectivity and conductance needs to be addressed by conditional expression of the mutant receptors. PMID- 11430907 TI - Agonist-stimulated [35S]GTPgammaS autoradiography: optimization for high sensitivity. AB - The receptor-stimulated accumulation of [35S]GTPgammaS provides a measure of functional coupling of G proteins with receptors. Sensitivity for autoradiographic visualization of [35S]GTPgammaS binding was improved two- to threefold in rat brain sections by optimizing assay conditions. Non-specific (NSB), basal and agonist-stimulated [35S]GTPgammaS binding were measured, using methadone, 5-carboxamidotryptamine and epinephrine for mu-opiate receptors, 5 HT(1A) receptors and alpha(2)-adrenoceptors. Basal and NSB were low in glycylglycine buffer compared to Tris or HEPES buffers, and agonist-stimulated [35S]GTPgammaS binding was more easily observed. Further optimization using glycylglycine buffer found increased signal-to-noise ratio with inclusion of dithiothrietol, increased [35S]GTPgammaS incubation time (2-4 h) and guanosine 5' diphosphate (GDP) preincubation (20-30 min), and use of [35S]GTPgammaS at 0.1 nM. Improved sensitivity was due to decreased NSB and basal [35S]GTPgammaS binding and agonist-stimulated binding were similarly affected for each receptor system. The assay conditions described should extend the use of agonist-stimulated [35S]GTPgammaS autoradiography to receptors, which produce low levels of [35S]GTPgammaS binding and to the measurement of changes in receptor-G protein coupling. PMID- 11430908 TI - Characterization of Ca(2+) channels involved in endothelin-1-induced mitogenic responses in vascular smooth muscle cells. AB - Ca(2+) channels involved in the endothelin-1-induced mitogenic response of cultured rat thoracic aorta smooth muscle cells, A7r5 cells, were characterized using the Ca(2+) channel blockers, LOE 908 and SK&F 96365. Stimulation of A7r5 cells with endothelin-1 induced a mitogenic response as well as a biphasic increase in the intracellular-free Ca(2+) concentration. Based on the sensitivity to nifedipine, a specific blocker of L-type voltage-operated Ca(2+) channel (VOCC), Ca(2+) influx through VOCC has a minor role in endothelin-1-induced mitogenic responses. On the other hand, Ca(2+) influx through voltage-independent Ca(2+) channels (VICCs) plays an important part in endothelin-1-induced mitogenesis. Moreover, based on their sensitivity to SK&F 96365 and LOE 908, VICCs consist of two types of Ca(2+)-permeable nonselective cation channels (designated NSCC-1 and NSCC-2) and a store-operated Ca(2+) channel (SOCC). Ca(2+) influx through NSCC-1, NSCC-2 and SOCC contributes to 35%, 30% and 35%, respectively, to the nifedipine-resistant component of the endothelin-1 mitogenic response. PMID- 11430909 TI - Stage-specific effects of a thiazolidinedione on proliferation, differentiation and PPARgamma mRNA expression in 3T3-L1 adipocytes. AB - To clarify the target phase of thiazolidinediones, which are ligands for peroxisome proliferator-activated receptor (PPAR)gamma, during adipocyte differentiation, the effects of a thiazolidinedione, pioglitazone, on every stage during the course of adipocyte differentiation were investigated. Pioglitazone did not affect the cellular protein content and [3H]thymidine incorporation into preconfluent 3T3-L1 preadipocytes. Induction of differentiation of confluent 3T3 L1 preadipocytes with insulin, dexamethasone and isomethylbutylxanthine for 48 h resulted in 30% inhibition of [3H]thymidine incorporation into the cells and 354% increase in cellular protein content. Pioglitazone at 1 microM accelerated the increase in cellular protein content by 33% and the inhibition in the [3H]thymidine incorporation by 12%. Pioglitazone, when added from the start of the induction stage, dose-dependently enhanced cellular triglyceride accumulation, and both basal and insulin-stimulated glucose transporting activity producing only a slight increase in the ratio of insulin stimulation to basal glucose transporting activity. In mature adipocytes, however, pioglitazone did not enhance either of the transporting activities. PPARgamma messenger RNA (mRNA) levels estimated by a semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) increased during the course of adipocyte differentiation. Although pioglitazone dose-dependently up-regulated PPARgamma mRNA levels in postconfluent preadipocytes without induction, it down-regulated them in mature adipocytes. Thus, a PPARgamma agonist, pioglitazone, arrested the growth, and increased protein content and PPARgamma mRNA levels in postconfluent preadipocytes, followed by commitment and hypertrophy of 3T3-L1 cells without changing insulin sensitivity, whereas it failed to stimulate glucose transporting activities and down-regulated PPARgamma mRNA expression in mature adipocytes. PMID- 11430910 TI - Effect of P-glycoprotein modulators on the human extraneuronal monoamine transporter. AB - The aim of this work was to investigate the effect of P-glycoprotein modulators on human extraneuronal monoamine transporter (EMT)-mediated transport. The experiments were performed using a cell line from human embryonic kidney (HEK293 cells) stably transfected with pcDNA3hEMT (293(hEMT)), or with pcDNA3 alone (293(control)). Of the P-glycoprotein modulators tested, rhodamine123, verapamil and daunomycin concentration-dependently inhibited EMT-mediated uptake of [3H]1 methyl-4-phenylpyridinium ([3H]MPP(+)). The corresponding IC(50)'s were found to be 3.6, 37 and 130 microM, respectively. By contrast, vinblastine, digitoxin and cyclosporine A were devoid of effect. The endogenous organic cation tyramine, but not choline, inhibited EMT-mediated transport (IC(50) of 468 microM). Moreover, L arginine and L-histidine (up to 1 mM) did not affect [3H]MPP(+) uptake. Finally, MPP(+) and tyramine trans-stimulated [3H]MPP(+) uptake, but rhodamine123 had no effect, and verapamil and daunomycin trans-inhibited [3H]MPP(+) uptake. In conclusion, this study shows that several cationic modulators of P-glycoprotein inhibit EMT-mediated transport. As a consequence, the interaction of P glycoprotein modulators with EMT must be taken into account, and the consequences of this interaction must not be forgotten when using such drugs in vivo. PMID- 11430911 TI - Trishomocubanes: novel sigma-receptor ligands modulate amphetamine-stimulated [3H]dopamine release. AB - Several trishomocubane analogues of the type 4-azahexacyclo [5.4.1.0(2,6).0(3,10).0(5,9).0(8,11)]dodecane exhibited moderate to high affinity at sigma-receptor subtypes and low or negligible affinity at dopamine and serotonin transporters (SERT). Selected compounds were examined for their effects on amphetamine-stimulated [3H]dopamine release from striatal slices in vitro. Compounds 1, 2, 3 and 4 significantly enhanced amphetamine-stimulated release in a concentration-dependent manner. Compound 4, with the highest affinity and selectivity for the sigma(2)-receptor subtype, displayed the greatest potency. The enhancement produced by 1 and 2 was fully reversed by the selective sigma(2) antagonists 1'-[4-[1-(4-fluorophenyl)-1-H-indol-3-yl]-1-butyl]spiro[iso benzofuran-1(3H), 4'piperidine] (Lu28-179), endo-N-(8-methyl-8 azabicyclo[3.2.1]oct-3-yl)-2,3-dihydro-(1-methyl)ethyl-2-oxo-1-H-benzimidazole-1 carboxyamidehydrochloride (BIMU-8) and the non-subtype selective antagonist N-[2 (3,4-dichlorophenyl)-ethyl]-N-methyl-2-pyrrolidinyl)ethylamine (BD1008). These data suggested a potential role for compounds 1 through 4 as sigma(2)-receptor agonists in functional studies. In addition, a D(3)-trishomocubane compound 5 displayed low affinity at sigma receptors (K(i)=3 microM) and moderate affinity at dopamine transporters (K(i)=623 nM). Compound 5 significantly inhibited the potentiation mediated by compound 2, presumably through sigma(2)-receptor antagonism, or a direct action on dopamine transporters. PMID- 11430912 TI - The role of transforming growth factor-beta and its receptors in human prostate smooth muscle cell fibronectin production. AB - In the present study, the role of transforming growth factor-beta (TGFbeta) on the production of the extracellular matrix component, fibronectin, in the prostate has been studied. The mRNA levels of fibronectin, TGFbeta and the two TGFbeta receptors, ALK5 (activin like kinase) and type II, were measured using reverse-transcription polymerase chain reaction (RT-PCR). TGFbeta increased fibronectin mRNA and protein (7-fold) in a concentration-dependent fashion. An interesting relationship between the two TGFbeta receptors was found in that TGFbeta caused an upregulation of its type I receptor mRNA (5-6-fold) and a downregulation of the type II receptor mRNA (5-fold). Time-course experiments revealed that the change in expression of the TGFbeta receptors reached maximum at 24 h with an early increase at 4-5 h, whereas the fibronectin gene expression was not significantly stimulated until about 24 h. These data provide evidence that TGFbeta stimulates extracellular matrix production in prostate cells. PMID- 11430913 TI - Ca(2+)-dependent sensitization of adenylyl cyclase activity. AB - It has been shown that intracellular Ca(2+) concentrations have multiple modulatory influences on hormone-stimulated adenylyl cyclase activity. Here, we report that increasing intracellular Ca(2+) concentration by treating cells with the Ca(2+) ionophore A23187 leads to a sensitization of the beta-adrenoceptor stimulated adenylyl cyclase activity in Ltk(-) cells expressing the human beta(2) adrenoceptor. The ionophore treatment led to a 20+/-5% increase of the maximal isoproterenol-stimulated cyclase activity that could be prevented by chelation of the extracellular Ca(2+) using EGTA. A similar Ca(2+)-mediated sensitization (20+/-6%) was observed when the adenylyl cyclase was directly stimulated by the diterpene forskolin indicating that the catalytic activity of the enzyme itself is modulated by the change in Ca(2+) concentration. Sensitization of both the isoproterenol- and forskolin-stimulated adenylyl cyclase activities were completely blocked by treatment with KN-62[1-[N,O-bis-(5-isoquinolinesulfonyl)-N methyl-L-tyrosyl]-4-phenylpiperazine], an inhibitor of the Ca(2+)-calmodulin dependent protein kinase (CamKinase). Taken together, our data reveal the existence of a CamKinase-dependent sensitization process acting at the level of the adenylyl cyclase catalytic moiety. PMID- 11430914 TI - Oxygen inhalation enhances striatal dopamine metabolism and monoamineoxidase enzyme inhibition prevents it: a microdialysis study. AB - In order to explore the effect of normobaric oxygen on the extracellular level of dopamine and its metabolites, oxygen (30, 60 and 90%) was administered to freely moving rats after the animals had been pretreated with either monoamineoxidase (MAO)-A and -B inhibitors (0.1 or 1 mg kg(-1) of clorgyline, 1 or 10 mg kg(-1) of selegiline and 75 mg kg(-1) pargyline) or control solution. The levels of dopamine and its metabolites were monitored in microdialysis samples collected every 20 min and directly applied to an on-line high-performance liquid chromatograph combined with electrochemical detection. Normobaric oxygen inhalation decreased the level of extracellular dopamine and increased that of 3,4-dihydroxyphenylacetic acid (DOPAC) in a concentration-dependent manner. These changes were partly prevented by pre-treatment with low doses of selegiline or clorgyline, i.e. by conditions in which monoamineoxidase-A or -B was inhibited. When both isoforms of monoamineoxidase were inhibited, there was a drastic increase in extracellular concentrations of dopamine and 3-methoxytyramine, and the levels of DOPAC and homovanilic acid (HVA) were very low. These results indicate that the intracellular metabolism of cytoplasmic dopamine is enhanced by normobaric hyperoxia in rat striatum. PMID- 11430915 TI - Lipopolysaccharide inhibits induction of long-term potentiation and depression in the rat hippocampal CA1 area. AB - We examined the effects of lipopolysaccharide, a bacterial endotoxin, on synaptic plasticity in the rat hippocampal CA1 area in vitro. Lipopolysaccharide suppressed the induction of long-term potentiation elicited by tetanic stimulation and long-term depression, elicited by low-frequency stimulation of Schaffer collateral-commissural fibres at 10 and 50 microg/ml, respectively. Lipid A (1 microg/ml), the biologically active component of lipopolysaccharide, mimicked the effects of 10 microg/ml lipopolysaccharide on long-term potentiation and depression. Nifedipine, an L-type voltage-sensitive Ca(2+) channel antagonist, did not influence the induction of long-term potentiation and depression, whereas a high concentration of extracellular calcium enabled long term potentiation induction in the presence of 10 microg/ml lipopolysaccharide. The NMDA receptor antagonist D,L-2-amino-5-phosphonovaleric acid (APV, 50 microM), nifedipine (10 microM) or lipopolysaccharide (10 or 50 microg/ml) partially reduced the magnitude of tetraethylammonium-induced long-term potentiation. Nifedipine combined with lipopolysaccharide completely blocked tetraethylammonium-induced long-term potentiation. Whole-cell voltage clamp recordings showed that lipopolysaccharide suppressed NMDA receptor-mediated excitatory postsynaptic currents (EPSCs). Our results indicate that lipopolysaccharide acutely modifies synaptic plasticity by blocking Ca(2+) entry through NMDA receptors, suggesting a possible mechanism for the amnesic action of bacterial infection. PMID- 11430916 TI - Induction of tolerance to the suppressant effect of the neurotensin analogue NT69L on amphetamine-induced hyperactivity. AB - Although several studies have indicated that neurotensin administered acutely has several pharmacological properties common with those of antipsychotic drugs, the effects of repeated exposure to neurotensin receptor agonism have been less well characterised. Here, we investigated the effect of the novel neurotensin-(8-13) analogue NT69L [(N-methyl-Arg), Lys, Pro, L-neo-Trp, tert-Leu, Leu] in animal models sensitive to central neurotensin receptor stimulation as well as in predictive models for antipsychotic activity and motor side-effect liability. Acute injection of NT69L (0.19-6.1 micromol/kg, s.c./i.p.) caused hypothermia (>2.5 degrees C) and reduction in spontaneous locomotor activity but failed to induce catalepsy. Furthermore, NT69L (0.10 micromol/kg, s.c.) counteracted the hyperlocomotion elicited by amphetamine (0.5 mg/kg, s.c.). However, repeated injections of NT69L (0.19 micromol/kg, s.c. for 6 days, twice daily) significantly reduced its effect on spontaneous locomotor activity and completely abolished its effect on amphetamine-elicited hyperactivity. Our data obtained after single injections of NT69L indicate that this drug stimulates central neurotensin receptors after peripheral administration and collectively support the notion that neurotensin receptor agonism is associated with an attractive pre clinical profile as regards both antipsychotic activity and motor side-effect liability. However, the present results also indicate that repeated neurotensin receptor stimulation may cause a desensitisation of neurotensin receptor mediated effects. PMID- 11430917 TI - Role of vanilloid VR1 receptor in thermal allodynia and hyperalgesia in diabetic mice. AB - We examined the role of the vanilloid VR1 receptor in the thermal hyperalgesia and allodynia seen in diabetic mice. Tail-flick latencies at source voltages of 35 and 50 V for a 50-W projection bulb in diabetic mice were significantly shorter than those in non-diabetic mice. Tail-flick latencies at 35 and 50 V in diabetic mice were increased by pretreatment with anti-vanilloid VR1 receptor serum. Intrathecal (i.t.) injection of anti-VR1 serum resulted in a significant increase in the tail-flick latency at 50 V in non-diabetic mice. However, i.t. pretreatment with anti-vanilloid VR1 receptor serum did not affect the tail-flick latency at a heat intensity of 35 V in non-diabetic mice. Thus, it seems likely that thermal allodynia and hyperalgesia in diabetic mice may be due to the sensitization of vanilloid VR1 receptors in primary sensory neurons in the spinal cord. PMID- 11430918 TI - Comparative effects of dextromethorphan and dextrorphan on morphine, methamphetamine, and nicotine self-administration in rats. AB - The effects of dextromethorphan and its metabolite dextrorphan on morphine, methamphetamine and nicotine self-administration and on responding for a nondrug reinforcer (water) were assessed in rats. Both dextromethorphan and dextrorphan decreased morphine self-administration at 10-30 mg/kg, s.c., decreased methamphetamine self-administration at 20 and 30 mg/kg, s.c., and decreased nicotine self-administration at 5-30 mg/kg, s.c.; doses of both drugs less than 40 mg/kg, s.c. did not affect responding for water. The equal potencies of dextromethorphan and dextrorphan suggest mediation of these effects by a non-NMDA receptor mechanism, possibly involving blockade of alpha3beta4 nicotinic receptors. The results also suggest that dextromethorphan should be tested extensively as a potential treatment for diverse populations of drug-abusing patients. PMID- 11430919 TI - Role of 5-HT(1B) receptors in the sensitization to amphetamine in mice. AB - The present study was designed to determine how 5-HT(1B) receptor ligands affected the development or the expression phase of sensitization to the amphetamine-induced locomotor response in mice. Mice were treated repeatedly (for 5 days) with amphetamine (2.5 mg/kg) in combination with either vehicle, N-[3-[3 (dimethylamino)ethoxy]-4-methoxyphenyl]-2'-methyl-4'-(5-methyl-1,2,4-oxadiazol-3 yl)-[1,1'-biphenyl]-4-carboxamide hydrochloride (SB 216641; an antagonist of 5 HT(1B) receptors), 3-(1,2,5,6-tetrahydro-4-pyridyl)-5-propoxypyrrolo[3,2 b]pyridine (CP 94,253; an agonist of 5-HT(1B) receptors), or SB 216641+CP 94,253; afterwards, on day 10, they received a challenge dose of amphetamine (2.5 mg/kg). In another experiment, mice were given either vehicle or amphetamine (2.5 mg/kg) for 5 days, and were then challenged with amphetamine (2.5 mg/kg) in combination with vehicle, SB 216641, or CP 94,253 on day 10. Locomotor hyperactivity induced by acute administration of amphetamine (day 1) was dose-dependently inhibited by SB 216641 and enhanced by CP 94,253, but not affected by a combination of SB 216641+CP 94,253. The 5-HT(1B) receptor ligands affected similarly the behavioral response to the challenge dose of amphetamine on day 10 (ca. 55-110% more potent than the response to its first administration) when they were combined with the psychostimulant during the development phase (days 1-5) of sensitization. On the other hand, neither SB 216641 nor CP 94,253 administered together with the challenge dose of amphetamine (day 10) affected its behavioral hyperactivity effect in mice treated repeatedly (days 1-5) with the psychostimulant alone. Our results suggest that 5-HT(1B) receptors may play a permissive role in the development, but not expression, of behavioral sensitization, as well as in the acute locomotor response to amphetamine in mice. PMID- 11430920 TI - Increases in ornithine decarboxylase activity in the positive inotropism induced by androgens in isolated left atrium of the rat. AB - It is well established that the intracellular receptors of androgens act as transcription factors upon their activation by androgen binding. However, a growing number of studies have associated androgens with rapid biological responses independent of their classical action mechanism. In this sense, 5alpha- and 5beta-dihydrotestosterone elicited a rapid positive inotropism in the isolated left atrium of the rat via cAMP-dependent mechanisms that may involve genomic effects. In addition, polyamines are mediators of several biological actions including those acute and long-term effects induced by androgens in the heart. The present study analyzed the role of polyamine synthesis in the cardiotonic effect of androgens in the left atrium of male Wistar rats, electrically stimulated (0.5 Hz, 5 ms and supramaximal voltage) and placed in an organ bath in 10 ml of Tyrode's solution. Incubation in the organ bath with an inhibitor of ornithine decarboxylase activity, alpha-difluoromethylornithine 10 mM, significantly decreased the positive inotropism induced by 5alpha- and 5beta dihydrotestosterone (0.1-100 microM). This suggests that ornithine decarboxylase seems to be involved in androgen-induced positive inotropism. Furthermore, 6-min exposure to 5alpha- or 5beta-dihydrotestosterone significantly increased the activity of ornithine decarboxylase from 61.81+/-7.53 (control) to 93.28+/-9.45 and 80.28+/-12 pmol/h/mg of protein, respectively. Northern blot analysis showed that 5alpha- and 5beta-dihydrotestosterone did not modify the level of expression of the ornithine decarboxylase gene. Therefore, our results suggest that polyamine synthesis might be involved in the positive inotropism elicited by androgens through the stimulation of ornithine decarboxylase activity without changes in the expression of the ornithine decarboxylase gene. PMID- 11430921 TI - T-1032, a novel specific phosphodiesterase type 5 inhibitor, increases venous compliance in anesthetized rats. AB - Nitric oxide (NO) donors including organic nitrates dilate capacitance vessels. As inhibition of phosphodiesterase type 5 results in the accumulation of guanosine 3'5'-cyclic monophosphate (cGMP), specific phosphodiesterase type 5 inhibitors are expected to have a vasodilator property similar to that of NO donors. To test this hypothesis, we examined the effect of methyl2-(4 aminophenyl)-1,2-dihydro-1-oxo-7-(2-pyridinylmethoxy)-4-(3,4,5-trimethoxyphenyl) 3-isoquinoline carboxylate sulfate (T-1032), a novel specific phosphodiesterase type 5 inhibitor, on mean arterial pressure and mean circulatory filling pressure (an index of venodilation) compared with that of nitroglycerin and diltiazem in mecamylamine- and noradrenaline-treated anesthetized rats. Intravenous infusion of T-1032 (0.1, 1, 10 microg/kg/min) dose-dependently decreased mean arterial pressure (-3.8+/-0.3%, -9.1+/-0.8%, -16.8+/-1.5% at doses of 0.1, 1 and 10 microg/kg/min, respectively) and mean circulatory filling pressure (-6.1+/-0.9%, 12.5+/-0.7%, -18.6+/-3.0% at doses of 0.1, 1 and 10 microg/kg/min, respectively). The mean circulatory filling pressure-mean arterial pressure relationship revealed that T-1032 had a selective action on the mean circulatory filling pressure compared with diltiazem (10, 100 microg/kg/min) and a similar or more selective effect than nitroglycerin (0.3, 3 and 30 microg/kg/min). In the next study, we calculated venous compliance and unstressed volume from the mean circulatory filling pressure-volume relationship. Intravenous infusion of T-1032 (3 microg/kg/min) increased venous compliance (3.35+/-0.40 in T-1032 vs. 2.31+/ 0.15 ml/kg/mm Hg in vehicle, P<0.05) without changing the unstressed volume (37.2+/-2.80 in T-1032 vs. 42.6+/-2.37 ml/kg in vehicle, P>0.05). It was concluded that T-1032 increased venous capacitance by increasing venous compliance, and that this selective phosphodiesterase type 5 inhibitor appeared to have a different vasodilator action from that of an NO donor and a Ca(2+) channel antagonist in that it had a selective action on the mean circulatory filling pressure. PMID- 11430922 TI - Metoprolol attenuates postischemic depressed myocardial function in papillary muscles isolated from normal and postinfarction rat hearts. AB - The present study was designed to test the hypothesis that metoprolol treatment may enhance tolerance to ischemia in normal and postinfarction rat myocardium. Myocardial infarction was induced by permanent ligation of the left coronary artery in adult rats. Animals were divided into sham-operated and infarction groups with or without metoprolol treatment. Metoprolol treatment (60 mg/kg/day via gastric gavage) was started on the second day after surgery and continued until sacrifice at 6 weeks after myocardial infarction. Isometric force and intracellular Ca(2+) ([Ca(2+)](i)) transients were simultaneously recorded in isolated left ventricular papillary muscles. Ischemia was simulated by immersing the muscles into fluorocarbon with hypoxia. Metoprolol treatment induced a significant improvement of isometric force and ameliorated diastolic [Ca(2+)](i) overload in postinfarction rat myocardium at baseline. Metoprolol treatment also reduced diastolic [Ca(2+)](i), ameliorated the depression of developed tension during ischemia, and enhanced recovery of postischemic depressed myocardial function in sham-operated and postinfarction rat papillary muscles. Protein levels of the sarcoplasmic reticulum Ca(2+) ATPase of left ventricles and postischemic papillary muscles from metoprolol-treated rats were higher than those in placebo-treated animals. We concluded, therefore, that metoprolol treatment produced appreciable improvement of intracellular Ca(2+) handling during ischemia-reoxygenation cycles, and enhanced recovery of postischemic depressed myocardial function in both normal and postinfarction rat myocardium. PMID- 11430923 TI - The alpha(1A)-adrenoceptor subtype mediates contraction in rat femoral resistance arteries. AB - In this study, alpha(1)-adrenoceptor subtypes were characterised in rat femoral resistance arteries mounted on a small vessel myograph. A-61603 was found to be more potent than noradrenaline and phenylephrine in these arteries. Brimonidine (UK 14304) could not evoke any contractile responses and the sensitivity to noradrenaline and phenylephrine was not affected by (8aR,12aS,13aS) 5,8,8a,9,10,11,12,12a,13a-decahydro-3-methoxy-12-(ethylsulphonyl)-6H-isoquino[2,1 g][1,6]-naphthyridine (RS 79948), ruling out the presence of alpha(2) adrenoceptors. Prazosin, 5-methyl-urapidil and 2-([2,6 dimethoxyphenoxyethyl]aminomethyl)-1,4-benzodioxane (WB 4101) produced rightward shifts in the sensitivity to noradrenaline, giving pA(2) values of 9.6, 9.4 and 10.4, respectively, in agreement with the presence of alpha(1A)-adrenoceptors. (8 [2-[4-(2-Methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4.5]decane-7,9-dione (BMY 7378; 1 microM) produced a small shift in the sensitivity of noradrenaline giving a pK(B) of 7.2. In the presence of 300 nM 5-methyl-urapidil, sensitivity to noradrenaline was not further shifted by 1 microM BMY 7378. Responses to noradrenaline were unaffected by the alpha(1B)-adrenoceptor alkylating agent chloroethylclonidine (1 microM). These results suggest alpha(1A)-adrenoceptors mediate contractile responses to noradrenaline in rat femoral resistance arteries. PMID- 11430924 TI - "Use-dependent" effects of cisapride on postrest action potentials in rabbit ventricular myocardium. AB - Repercussions of cisapride-induced blocking effects on repolarisation of K(+) channels in open and inactivated states investigated in rabbit ventricular myocardium during rest and under stimulation were compared with effects of K(+) blocking drugs (4-aminopyridine, dofetilide, terikalant). Major lengthening in the first postrest action potential indicates affinity for closed channels. Gradual lengthening during stimulation implies affinity for open channels. Four (control, add-in, steady-state, washout) 20-min rest periods were alternated with regular stimulation (0.5 Hz). Each drug was added during add-in and steady-state periods. Similarly to dofetilide (10 nM) and terikalant (0.3 microM), cisapride (1 microM) increasingly lengthened action potentials during stimulation, whereas 4-aminopyridine (1 mM) prolonged mostly the first postrest action potential. Our results indicate that cisapride induced use-dependent lengthening of repolarisation, compatible with an affinity for open K(+) channels. We also found that in isolated rabbit ventricular myocytes, cisapride (1-10 microM) decreased the inward rectifier K(+) current, an effect contributing to the proarrhythmic potential. PMID- 11430925 TI - Effects of cilnidipine on nitric oxide and endothelin-1 expression and extracellular signal-regulated kinase in hypertensive rats. AB - We evaluated the effects of cilnidipine, a long-acting Ca(2+) channel antagonist, on endothelial nitric oxide synthase (eNOS), preproendothelin-1 and endothelin ETA receptor expression in the left ventricle, and evaluated the relations between these effects and coronary microvascular remodeling and extracellular signal-regulated kinases belonging to one subfamily of mitogen-activated protein kinases in deoxycorticosterone acetate (DOCA)-salt hypertensive rats. Cilnidipine (DOCA-cilnidipine, 1 mg/kg/day, subdepressor dose) or vehicle (DOCA-vehicle) was given after induction of DOCA-salt hypertension for 5 weeks. The eNOS mRNA and protein expression in the left ventricle was significantly lower in DOCA-vehicle than in control rats and significantly higher in DOCA-cilnidipine than in DOCA vehicle rats. Preproendothelin-1 and endothelin ETA receptor expression levels and phospho-p42/p44 extracellular signal-regulated kinase activities were significantly increased in DOCA-vehicle compared with control rats and significantly suppressed in DOCA-cilnidipine compared with DOCA-vehicle rats. DOCA-vehicle rats showed a significant increase in the wall-to-lumen ratio, perivascular fibrosis and myocardial fibrosis, with all these parameters being significantly improved by cilnidipine. These results led us to conclude that phospho-p42/p44 extracellular signal-regulated kinase activities may contribute to the coronary microvascular remodeling of DOCA rats and that protective effects of cilnidipine on cardiovascular remodeling may be at least in part mediated by an increased eNOS expression and a decreased endothelin-1 and endothelin ETA receptor expression in the left ventricle. PMID- 11430926 TI - Magnolol reduces myocardial ischemia/reperfusion injury via neutrophil inhibition in rats. AB - The accumulation of oxygen-free radicals and activation of neutrophils are strongly implicated as important pathophysiological mechanisms mediating myocardial ischemia/reperfusion injury. It has been proven that various antioxidants have cardioprotective effects. Magnolol, an active component extracted from the Chinese medicinal herb Magnolia officinalis, possesses potent antioxidant and free radical scavenging activities. In this study, the cardioprotective activity of magnolol was evaluated in an open-chest anesthetized rat model of myocardial ischemia/reperfusion injury. The results demonstrated that pretreatment with magnolol (0.2 and 0.5 microg/kg, i.v. bolus) at 10 min before 45 min of left coronary artery occlusion, significantly suppressed the incidence of ventricular fibrillation and mortality when compared with the control group. Magnolol (0.2 and 0.5 microg/kg) also significantly reduced the total duration of ventricular tachycardia and ventricular fibrillation. After 1 h of reperfusion, pretreatment with magnolol (0.2 and 0.5 microg/kg) caused a significant reduction in infarct size. In addition, magnolol (0.2 microg/kg) significantly reduced superoxide anion production and myeloperoxidase activity, an index of neutrophil infiltration in the ischemic myocardium. In addition, pretreatment with magnolol (0.2 and 0.5 microg/kg) suppressed ventricular arrhythmias elicited by reperfusion following 5 min of ischemia. In vitro studies of magnolol (5, 20 and 50 microM) significantly suppressed N-formylmethionyl leucyl-phenylalanine (fMLP; 25 nM)-activated human neutrophil migration in a concentration-dependent manner. It is concluded that magnolol suppresses ischemia and reperfusion-induced ventricular arrhythmias and reduces the size of the infarct resulting from ischemia/reperfusion injury. This pronounced cardioprotective activity of magnolol may be mediated by its antioxidant activity and by its capacity for neutrophil inhibition in myocardial ischemia/reperfusion. PMID- 11430927 TI - Effects of methylenedioxymethamphetamine on noradrenaline-evoked contractions of rat right ventricle and small mesenteric artery. AB - We have compared the effects of methylenedioxymethamphetamine (MDMA) and cocaine on contractions to noradrenaline in 1 Hz paced rat right ventricular strips, and in rat small mesenteric artery and aorta. Noradrenaline increased the force of contraction of 1 Hz paced ventricular strips with a pD(2) (-log EC(50)) of 5.64+/ 0.07. Both cocaine (10 microM) and MDMA (10 microM) significantly increased the potency of noradrenaline to 6.31+/-0.11 and 6.42+/-0.13, respectively. However, in the presence of cocaine (10 microM) which increased the potency of noradrenaline to 6.78+/-0.15, MDMA (10 microM) no longer increased the potency of noradrenaline (pD(2) of 6.78+/-0.32). Likewise, following chemical sympathectomy, MDMA failed to increase the potency of noradrenaline. The potency of the agonist isoprenaline, which is not a substrate for the noradrenaline transporter, was not increased by either cocaine or MDMA. In rat small mesenteric artery, but not aorta, MDMA and cocaine significantly increased the potency of noradrenaline, but in the presence of cocaine, MDMA had no further effect. Hence, MDMA shares with cocaine an ability to potentiate the actions of noradrenaline, an action in the case of MDMA which may involve competitive blockade of the noradrenaline transporter, rather than simply displacement of noradrenaline. Since cocaine is linked to an increased incidence of myocardial infarction, these results may have implications in terms of cardiac morbidity of MDMA. PMID- 11430928 TI - Nebivolol, carvedilol and metoprolol do not influence cardiac Ca(2+) sensitivity. AB - It has been argued that some beta-adrenoceptor antagonists may directly influence myofibrillar cross-bridge interaction in cardiac skinned fiber preparations of animal models. The present study investigates the effects of nebivolol, metoprolol and carvedilol on tension development of Triton X-100 skinned fibers obtained from human failing myocardium as well as on force of contraction and intracellular Ca(2+) transient in isolated trabeculae. In skinned fiber preparations, none of the beta-adrenoceptor antagonists (10 microM) influenced Ca(2+) sensitivity of tension development or maximal Ca(2+) activated tension (DT(max)): control: EC(50) for Ca(2+): 1.28+/-0.05 microM, DT(max): 14.09+/-0.59 mN/mm(2); nebivolol: 1.36+/-0.1 microM, 14.14+/-0.95 mN/mm(2); carvedilol: 1.32+/ 0.11 microM, 13.83+/-0.90 mN/mm(2); metoprolol: 1.34+/-0.14 microM, 13.72+/-0.36 mN/mm(2). Simultaneous measurement of force and Ca(2+) transient in the presence of the beta-adrenoceptor antagonists (3 microM) showed that the decrease in force of contraction was paralleled by a similar decrease in the intracellular Ca(2+) transient. In conclusion, none of the investigated beta-adrenoceptor antagonists influenced Ca(2+) sensitivity of myofibrillar tension development in human failing myocardium. PMID- 11430929 TI - K(ATP) channels predominantly regulate conduit vessel tone in normoxic cat pulmonary arteries in vivo. AB - Through our investigations of the intact pulmonary circulation, we aimed to find out whether K(ATP) channels contribute to regulating basal vascular tone and to clarify which vascular segments dilate during K(ATP) channel activation under basal tone conditions. Using an X-ray television system on anesthetized cat lungs, we measured internal diameter (ID) responses to two K(ATP) channel inhibitors (glibenclamide and 4-morpholinecarboximidine-N-1-adamantyl-N' cyclohexyl-hydrochloride (U-37883A)) and to an activator (levcromakalim) in normoxic pulmonary arteries. In conduit arteries (800-3000 microm ID), the inhibitors and activator induced larger ID constrictions (14-17%) and dilatations (29-32%), respectively. However, in resistance arteries (<500 microm), the constriction response was negligible and the dilatation response relatively small (5-10%). The data suggest that K(ATP) channels are active and capable of regulating basal vascular tone primarily within conduit pulmonary arteries even though these channels are present in all pulmonary arteries. PMID- 11430930 TI - Genital grooming and emesis induced by vanilloids in Suncus murinus, the house musk shrew. AB - The potential of resiniferatoxin and capsaicin to modulate emesis and genital grooming was investigated in Suncus murinus. Resinifertoxin (3-30 nmol, i.c.v.), E-capsaicin (10-100 nmol, i.c.v.) and Z-capsaicin (100 nmol, i.c.v.) induced emesis (P<0.05) and subsequently antagonised the emetic response induced by intragastric copper sulphate (480.6 micromol/kg; P<0.05). However, resiniferatoxin failed to affect nicotine-induced (30.7 mol/kg, s.c.) emesis (P>0.05). Only resiniferatoxin induced genital grooming that was antagonised (P<0.05) by capsazepine (300-600 nmol, i.c.v.) and ruthenium red (3 nmol, i.c.v.). E-capsaicin-induced emesis was antagonised by capsazepine (300-600 nmol, i.c.v.; P<0.05) and ruthenium red (3 nmol, i.c.v.; P<0.05) but resiniferatoxin induced emesis was resistant to capsazepine (30-600 nmol, i.c.v.; P>0.05). The emetic action of resiniferatoxin but not E-capsaicin was subject to tachyphylaxis. In cross-tachyphylaxis experiments, E-capsaicin reduced the genital grooming induced by resiniferatoxin (P<0.05). The data are discussed in relation to the classification of vanilloid receptors and mechanisms involved in emesis and genital grooming. PMID- 11430931 TI - Nucleotide-evoked relaxation of rat vas deferens--a possible role for endogenous ATP released upon alpha(1)-adrenoceptor stimulation. AB - The possibility was tested that endogenous ATP released upon alpha(1) adrenoceptor activation causes relaxation of the rat vas deferens smooth muscle. ATP, 2-methylthio ATP and adenosine relaxed the vas deferens precontracted with 80 mM K(+). The metabolically stable P2 receptor agonists alpha,beta-methylene ATP (alpha,beta-MeATP) and adenosine 5'-O-(2-thiodiphosphate) (ADPbetaS) had little or no effect. The adenosine P1 receptor antagonist 8-(para sulfophenyl)theophylline did not significantly affect the response to ATP. The P2 receptor antagonist reactive blue 2 markedly reduced the relaxation (by up to 73%); suramin, pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) and acid blue 129 caused no change. ATP, but not alpha,beta-MeATP, also attenuated contractions elicited by noradrenaline at resting tension; reactive blue 2 blocked the inhibitory effect of ATP. Reactive blue 2, by itself, enhanced the response to noradrenaline (by up to 36%); suramin, PPADS and acid blue 129 caused no change. In the presence of the ATP-degrading enzymes apyrase and nucleotide pyrophosphatase, the facilitatory effect of reactive blue 2 was lost. Apyrase, by itself, enhanced the response to noradrenaline (by 13%). The results indicate that endogenous ATP, released from rat vas deferens smooth muscle upon alpha(1) adrenoceptor stimulation, causes relaxation. The site of action of ATP is not a typical smooth muscle P2Y receptor. PMID- 11430932 TI - Beta(3)-adrenoceptors control Cl(-) conductance in rabbit nasal epithelium. AB - We have investigated the effects of beta(3)-adrenoceptor stimulation in vivo on nasal epithelium. We have recorded the transepithelial potential difference in New Zealand white rabbit nostrils. Superfusion of the nasal epithelial surface with a Cl(-)-free medium supplemented with amiloride, hyperpolarized the nasal potential difference. Isoprenaline produced a hyperpolarization of the nasal potential difference that was not prevented by nadolol, a potent beta(1)-/beta(2) adrenoceptor antagonist, but was abolished by bupranolol, a nonselective beta(1 3)-adrenoceptor antagonist. SR 58611 ((RS)-N-[(25)-7-ethoxycarbonylmethoxy 1,2,3,4-tetrahydronapht-2-yl]-(2R)-2-(3-chlorophenyl)-2 hydroethanamine hydrochloride) and CGP 12177 (4-[3-t-butylamino-2-hydroxypropoxy]benzimidazol-2 1), a preferential and a partial beta(3)-adrenoceptor agonists, respectively, also produced hyperpolarization of the nasal potential difference. SR 59230 (3-(2 ethylphenoxy)-1-[(1S)1,2,3,4-tetrahydronaphth-1-ylaminol]-(2S)-2-propanol oxalate), a selective beta(3)-adrenoceptor antagonist, abolished the effects of CGP 12177. We conclude that beta(3)-adrenoceptor stimulation resulted in modifications in the nasal potential difference. These findings strengthen the view that beta(3)-adrenoceptors are implicated in controlling water and salt transport in the normal respiratory epithelium. PMID- 11430933 TI - In vitro effects of E3040, a dual inhibitor of 5-lipoxygenase and thromboxane A(2) synthetase, on eicosanoid production. AB - In vitro pharmacological profiles of E3040, 6-hydroxy-5, 7-dimethyl-2 (methylamino)-4-(3-pyridylmethyl) benzothiazole were investigated. Against the 5 lipoxygenase activity of rat basophilic leukemia cells, E3040 and zileuton (a 5 lipoxygenase inhibitor) had an IC(50) of 0.23 and 0.93 microM, respectively. Against the thromboxane A(2) synthetase activity of human platelets, E3040 had an IC(50) of 0.01 microM, which was comparable to that of OKY-1581 (sodium (E)-3-[4 (3-pyridylmethyl) phenyl]-2-methylacrylate, a thromboxane A(2) synthetase inhibitor). Against cyclooxygenase activity of sheep seminal vesicles, E3040 showed no inhibition (IC(50), >300 microM). Sulfasalazine and 5-aminosalicylic acid, therapeutic drugs for inflammatory bowel disease, inhibited 5-lipoxygenase activity with an IC(50) of 293 and 970 microM, respectively. Sulfasalazine inhibited thromboxane A(2) synthetase activity with an IC(50) of 20 microM. In rat peritoneal leukocytes, E3040 inhibited leukotriene B(4) and thromboxane B(2) production with an IC(50) of 0.17 and 0.24 microM, respectively. E3040 inhibited leukotriene B(4) production in human neutrophils and thromboxane B(2) production in human platelets (IC(50) of 0.21 and 0.09 microM, respectively). These results indicated that E3040 potently inhibited 5-lipoxygenase and thromboxane A(2) synthetase and blocked leukotriene B(4) and thromboxane B(2) production in rat peritoneal and human blood cells. PMID- 11430934 TI - Pregnancy following uterine fibroid embolization. AB - OBJECTIVE: This paper seeks to evaluate the ability to deliver term pregnancies following uterine fibroid embolization, and to identify impediments to pregnancy in the embolization procedure. STUDY DESIGN: Four physicians performed embolization procedures at various facilities. Patients were asked if fertility was an issue prior to embolization. We measured follicle-stimulating hormone levels before and after embolization. Clinical follow-up, six months following embolization was obtained by interview. Patients were questioned regarding attempts to conceive, menstrual history, and subsequent pregnancy. MAIN OUTCOME MEASURES: Complications were calculated upon the entire patient population, whether or not fertility was identified as a goal. Fertility risks from embolization were identified. We measured radiation exposure in a random consecutive group of 50 women undergoing embolization. All patients who conceived were asked the details of the pregnancy. RESULTS: Four hundred women underwent uterine fibroid embolization between 1996 and 1999. One hundred and thirty nine patients stated a desire for fertility after embolization. Of these, 52 were <40 years old. Seventeen pregnancies have been reported in 14 women. Five spontaneous abortions were observed. Ten women have had normal term deliveries and two women are currently pregnant. No perfusion problems, either during the pregnancy or labor, were reported. The average radiation dosage calculated for 50 women undergoing embolization was 14 rads. Four women under 45 years old suffered premature menopause (10/1000). Two women underwent hysterectomy as a complication of embolization (5/1000). CONCLUSION: The risks of infertility following embolization, premature menopause, and hysterectomy are small, as is the radiation exposure during embolization. These risks compare favorably with those associated with myomectomy. Fertility rates appear similar to patients undergoing myomectomy. No problems, either during pregnancy or delivery, have been observed after embolization. The course of pregnancy and delivery was normal after embolization with no maternal or fetal complications reported. These findings confirm results from other centers. Desire for future pregnancy is not a contraindication to fibroid embolization. PMID- 11430935 TI - Complications associated with cesarean section in HIV-infected patients. AB - OBJECTIVE: To document complications associated with cesarean section in HIV infected women. METHOD: A cross-sectional study was done on 307 women who delivered by cesarean section. All these patients, irrespective of their HIV status, were managed according to the same preset protocol. RESULT: Of the 307 women, 59 (19%) were HIV-positive. The only significant differences were a higher prevalence of positive serology for syphilis (32% vs. 12%, P=0.0014) as well as more cases of post-operative endometritis in the HIV-positive group (24% vs. 7%, P=0.0003). Post-operative endometritis was documented in 44% of HIV-infected patients with a CD4 count below 400. However, the severity of endometritis, measured by antibiotic treatment and duration of hospital stay, was similar in the two groups. Overall, there was no difference in the duration of hospital stay, post-operative anemia, wound infection, birth weight and perinatal mortality between the two groups. CONCLUSION: HIV-infected patients undergoing cesarean section are at increased risk for post-operative endometritis, but the severity of the infection is not different from that in HIV-negative patients. PMID- 11430936 TI - Supplementary calcium in prevention of pre-eclampsia. AB - OBJECTIVE: To study the effect of calcium supplementation on reduction of pre eclampsia in Iranian women at high risk of pre-eclampsia. METHODS: Thirty pregnant women at high risk of developing pre-eclampsia were randomized to 2 g of daily calcium and placebo. Subjects and investigators were blinded to treatment allocation. The inclusion criteria were positive rollover test, having at least one risk factor for pre-eclampsia, between 28 and 32 weeks of pregnancy, and blood pressure less than 140/90 (mm Hg). Exclusion criteria were having any chronic condition. Pre-eclampsia was defined as systolic/diastolic blood pressure over 140/90 plus proteinuria. All the subjects were followed up to delivery. RESULT: A sevenfold reduction in the occurrence of pre-eclamsia were seen among the calcium group compared with the placebo group (7 out of 15 developed pre eclampsia in control group vs. 1 out of 15 in calcium group, P=0.014). No major effect on pregnancy induced hypertension was seen (66% in placebo and 76% in calcium group developed hypertension) but the onset of hypertension was delayed 3 weeks in the calcium group. A longer duration of pregnancy was seen in calcium than placebo (mean+/-S.D.=37+/-2 for calcium and 34+/-2 for placebo, P<0.05). Infants born to the calcium group, on average, were 552 g heavier than infants born to the placebo group, P<0.05. CONCLUSION: Calcium supplementation is beneficial for preventing pre-eclampsia among Iranian women at high risk of developing pre-eclampsia. PMID- 11430937 TI - Anesthesia in pregnant women with HELLP syndrome. AB - OBJECTIVE: Our purpose was to determine the types of anesthesia and neurologic or hematologic complications found in pregnancy with HELLP syndrome providing analgesia for cesarean section. METHODS: This is a retrospective study. For the period of 1 July 1996, through 30 June 2000, we reviewed the charts of all patients with HELLP syndrome who had cesarean section. RESULTS: During the period of study 119 patients had HELLP syndrome. Eighty-five patients had cesarean delivery and 34 had vaginal delivery. Seventy-one patients had diagnosed HELLP syndrome previous to the anesthesia and 14 postcesarean delivery; the range platelet count was 19000-143000/microl. Of these 71, 58 had an epidural anesthesia, 9 had general anesthesia and 4 had spinal anesthesia. There were no neurologic complications or bleeding in the epidural space. CONCLUSION: We found no documentation of any neurologic or hematologic complications of women with HELLP syndrome and neuraxial anesthesia. PMID- 11430938 TI - Subtotal hysterectomy for uterine rupture. AB - OBJECTIVE: The aim of this paper was to stratify patients with uterine rupture in extremis (with hypotension, change in vital organ function, and altered mental status) and evaluate outcome for patients undergoing subtotal hysterectomy (STH) vs. uterine repair (UR). METHODS: Of 39667 hospital deliveries between 1993 and 1998 at a university hospital, 367 presented with uterine rupture. Of these, 96 presented in extremis and had an irregularly torn uterus found at operation. Charts were retrospectively reviewed to stratify patient population with uterine rupture and their outcome. Results were analyzed using Fisher's exact test. RESULTS: Patients with uterine rupture were 31-35 years old, para> or =3, had received no antenatal care, and presented with rupture in the lower uterine segment. Sixty-one patients underwent STH while 35 underwent UR. The groups were similar with respect to patient age, parity, cause of rupture, and clinical condition at the time of evaluation (P=0.2). The operative time for STH was significantly less than UR, 35 min vs. 75 min (P<0.01). Maternal mortality was significantly higher in patients undergoing UR vs. STH, 46% vs. 20% (P=0.01). Maternal morbidity occurred in 30% of patients undergoing STH vs. 50% in patients undergoing UR (P=0.01). The time to discharge was significantly less in patients undergoing STH compared to UR, 14.5 days vs. 27 days (P<0.01). CONCLUSIONS: Patients with uterine rupture in extremis tend to be young, multiparous, receive no antenatal care, and have uterine rupture of the lower segment. STH significantly lowered operative time, morbidity, time to discharge, and mortality than UR in patients in extremis with uterine rupture. PMID- 11430939 TI - Efficacy of 7-day treatment with metronidazole+miconazole (Neo-Penotran) - a triple-active pessary for the treatment of single and mixed vaginal infections. AB - OBJECTIVE: To evaluate the efficacy of Neo-Penotran pessaries (metronidazole 500 mg + miconazole nitrate 100 mg) in candidal, bacterial and trichomonal vaginitis and in mixed vaginal infections. METHOD: Ninety-seven patients with clinical diagnosis of vaginitis entered this open, non-comparative study. Each patient inserted one pessary twice daily for 7 days. Gynecological and microbiological assessments were carried out before, and 8-10/21-23 days after the start of treatment. RESULTS: Vaginitis symptoms were resolved in 91% of the 74 patients evaluated, and improved in a further 7%. Microbiological cure rates were 97.3% for trichomonal, 86.6% for bacterial and 81% for candidal vaginitis. Recurrence rates were 2.7, 3.8 and 16.1%, respectively. Overall microbiological cure rate for mixed infections was 86%, with 93% for trichomonal+bacterial, and 73% for bacterial+candidal vaginitis. In two out of three cases with trichomonal+bacterial+candidal infection, the microorganisms were eradicated completely. CONCLUSION: Neo-Penotran provides immediate and effective treatment for vaginitis, irrespective of single or multiple infection, even when the diagnosis may be uncertain. PMID- 11430940 TI - Relationship between age, histological type, and size of ovarian tumors. AB - OBJECTIVE: To clarify the relationship between age, histological type, and size of ovarian tumors. METHOD: A review was made of 1648 cases of histopathologically diagnosed ovarian tumors and tumor-like lesions, and information on the age of the patients and size of the tumor was obtained. Statistical analysis was performed using Kruskal-Wallis tests or Mann-Whitney U-tests. RESULTS: There were 840 (51%) cases of benign tumors, 73 (4%) cases of tumors of low malignant potential (LMP), 268 (16%) cases of malignant tumors and 467 (28%) cases of tumor like lesions. The age of the patients was significantly different among tumor like lesions (34.6+/-8.1 years), benign tumors (39.8+/-16.4 years), LMP tumors (45.2+/-18.3 years) and malignant tumors (51.9+/-13.0 years) (P<0.0001). The maximum diameter of the tumors was significantly different among tumor-like lesions (7.1+/-3.3 cm), benign tumors (10.9+/-5.6 cm), malignant tumors (13.6+/ 6.5 cm) and LMP tumors (18.5+/-6.8 cm) (P<0.0001). CONCLUSION: The distribution of tumor histological type (tumor-like lesions, benign, LMP and malignant) was correlated against patient age and lesion diameter, with tumors in older patients or larger tumors more likely to be malignant. PMID- 11430941 TI - The management of the second stage of labor. AB - OBJECTIVE: To review obstetric practice in a single maternity hospital with respect to the assisted vaginal delivery rate. METHOD: A retrospective analysis of the hospital statistics, labor ward records, casenotes and management protocols with respect to the second stage of labor was performed in a single maternity unit in the UK; the study included 43352 women who delivered a baby between the years of 1987 and 1997. The study looked at the rate of assisted vaginal deliveries, cesarean section, epidural in labor, and as well as the perinatal mortality rate. RESULTS: In the 11-year period of the study, 43352 women delivered with a mean assisted vaginal delivery rate of 3.70%, cesarean section rate of 12.4% and an epidural rate of 31.5%. CONCLUSION: The management of the second stage of labor in this unit results in an assisted vaginal delivery rate significantly below the national average. The authors highlight the paucity of research in this important area of practice. PMID- 11430942 TI - Facial and body blood contamination in vaginal delivery. PMID- 11430943 TI - Aberrant uterine artery as a cause of uterine artery embolization treatment failure. PMID- 11430944 TI - Estrogen 'add-back' and lipid profile during GnRH agonist (triptorelin) therapy. PMID- 11430945 TI - Mammographic density changes in postmenopausal women using tibolone therapy. PMID- 11430948 TI - Transvaginal ultrasound and digital examination in predicting successful labor induction. AB - OBJECTIVE: To compare transvaginal ultrasound and digital cervical examination in predicting successful induction in post-term pregnancy. METHODS: Transvaginal ultrasound and digital vaginal examinations were performed on 122 women at 41 or more weeks' gestation, immediately before labor induction. Ultrasound assessments of cervical length, dilatation, and presence of funneling were compared with the components of the Bishop score. The primary outcome was the rate of vaginal delivery. Secondary outcomes assessed included the rates of active labor in 12 hours, vaginal delivery in 12 and 24 hours, mean duration of latent phase, and induction to vaginal delivery interval. Linear and multiple logistic regression models were generated to identify factors independently associated with successful induction. RESULTS: No ultrasound characteristic predicted primary or secondary outcomes. Bishop score (odds ratio [OR] 2.98, 95% confidence interval [CI] 1.71, 5.20), cervical position (OR 4.35, 95% CI 1.41, 12.50), and maternal age (OR 1.15, 95% CI 1.01, 1.30) independently predicted vaginal delivery. Maternal weight (OR 0.96, 95% CI 0.94, 0.98), cervical dilatation (OR 6.08, 95% CI 1.70, 21.68), and effacement (OR 2.34, 95% CI 1.16, 4.73) independently predicted active labor in 12 hours. Independent predictors of vaginal delivery in 12 hours were induction method (P <.001), cervical dilatation (OR 11.16, 95% CI 3.17, 39.29), gravidity (OR 2.06, 95% CI 1.13, 3.77), and maternal weight (OR 0.96, 95% CI 0.93, 0.99). Cervical effacement (OR 2.70, 95% CI 1.59, 4.57) and parity (OR 7.10, 95% CI 2.22, 22.72) independently predicted vaginal delivery in 24 hours. Maternal weight, cervical position, and cervical dilatation were independently associated with latent phase labor duration. Factors independently associated with length of induction to delivery interval were parity, cervical effacement, and maternal weight. CONCLUSION: Transvaginal ultrasound does not predict successful labor induction in post-term pregnancy as well as digital cervical examination. PMID- 11430949 TI - Group B streptococcal disease prevention practices of obstetrician-gynecologists. AB - OBJECTIVE: To describe group B streptococcal (GBS) disease prevention practices of obstetrician-gynecologists. METHODS: We surveyed 1019 ACOG Fellows-the 419 members of the Collaborative Ambulatory Research Network (CARN) and 600 randomly selected non-CARN Fellows. RESULTS: There were 601 eligible respondents. More than 95% in both the CARN and the non-CARN groups reported adopting one of three GBS prevention strategies. The most commonly reported strategy was a combination approach not described in the consensus guidelines. The second most common strategy was the screening-based strategy; the risk-based strategy was third. Most respondents provided GBS information to all prenatal patients, but those using a risk-based strategy and those in solo practice were less likely to do so. Less than 60% in each group used penicillin as their first choice for GBS prophylaxis. More than 20% in each group who routinely screened for GBS did not collect both vaginal and rectal cultures. Respondents rated ACOG publications as the most important influence on their GBS prevention approach. CONCLUSION: Almost all ACOG Fellows have adopted a GBS prevention strategy. The importance of providing GBS prevention information to all patients, use of penicillin, and collection of both vaginal and rectal cultures should be reinforced. PMID- 11430950 TI - Varicella vaccine exposure during pregnancy: data from the first 5 years of the pregnancy registry. AB - OBJECTIVE: To assess the risks of congenital varicella syndrome and other birth defects in offspring of women who inadvertently received varicella vaccine during pregnancy or within 3 months of conception. METHODS: Pregnant women inadvertently exposed to varicella vaccine, reported voluntarily, were enrolled in the Pregnancy Registry for VARIVAX (Merck & Co., Inc., West Point, PA). The pregnancies were monitored and the outcomes ascertained from questionnaires completed voluntarily by the health care providers. The rates of congenital varicella syndrome and congenital anomalies were calculated for seronegative women prospectively reported to the registry. RESULTS: From March 17, 1995 through March 16, 2000, 362 pregnancy outcomes were identified from prospective reports. Ninety-two women were known to be seronegative to varicella, of whom 58 received their first dose of vaccine during the first or second trimester. No cases of congenital varicella syndrome were identified among 56 live births (rate 0%, 95% confidence interval [CI] 0, 15.6). Among all the prospective reports of live births, five congenital anomalies were reported. No specific pattern was identified in either the susceptible cohort or the sample population as a whole. CONCLUSION: No abnormal features have been reported that suggested the occurrence of congenital varicella syndrome or other birth defects related to vaccine exposure during pregnancy. Because of the small numbers, this study has limited precision, so continued surveillance is warranted. However, these results should provide some assurance to health care providers and women with inadvertent exposure before or during pregnancy. PMID- 11430951 TI - Association of maternal fever during labor with neonatal and infant morbidity and mortality. AB - OBJECTIVE: To examine the association of intrapartum fever with infant morbidity and early neonatal (0-6 days) and infant (0-364 days) death. METHODS: We carried out a retrospective cohort analysis among singleton live births in the United States for the period 1995-1997 using the National Center for Health Statistics linked birth-infant death cohort data. RESULTS: Among the 11,246,042 singleton live births during the study period, intrapartum fever (at least 38C) was recorded in 1.6%. Intrapartum fever was associated with early neonatal (adjusted odds ratio [OR], 95% confidence interval [CI] for preterm and term infants respectively: 1.32; 1.11, 1.56 and 1.67; 1.14, 2.46) and infant (OR, 95% CI for preterm and term, respectively: 1.31; 1.14, 1.51 and 1.27; 1.01, 1.59) death among nulliparous mothers. Among preterm infants of parous mothers, intrapartum fever was associated with early neonatal (OR 1.29, 95% CI 1.01, 1.64) death. In the combined analyses (infants of nulliparous and parous mothers), intrapartum fever was a strong predictor of infection-related death. These associations were stronger among term (OR 3.16, 95% CI 1.56, 6.40 for early neonatal; OR 1.75, 95% CI 1.20, 2.57 for infant death) than preterm infants (OR 1.52, 95% CI 1.15, 2.00 for early neonatal; OR 1.29, 95% CI 1.05, 1.57 for infant death). Intrapartum fever was also a risk factor for meconium aspiration syndrome, hyaline membrane disease, neonatal seizures, and assisted ventilation. CONCLUSION: Intrapartum fever is an important predictor of neonatal morbidity and infection-related mortality. PMID- 11430952 TI - Uterine artery embolization for leiomyomata. AB - OBJECTIVE: To determine whether uterine artery embolization is safe and effective for treating uterine leiomyomata. METHODS: We analyzed 200 consecutive patients (61 reported previously) undergoing uterine artery embolization for the treatment of uterine leiomyomata at a single institution. After treatment, follow-up data were obtained by written questionnaire mailed to the patients at intervals of 2 weeks, 3 months, 6 months, and 12 months after treatment. Follow-up imaging was obtained at 3 months and 12 months after therapy. All complications and subsequent gynecologic interventions were recorded prospectively, obtained using the patient questionnaires and physician contact. The percentages and their 95% confidence intervals (CI) were calculated to compare the symptoms at follow-up. Proportional odds models for repeated ordinal responses were used to assess the stability of symptom improvement over time. RESULTS: The mean follow-up was 21 months (minimum 12). Heavy menstrual bleeding improved in 87% (95% CI 82%, 92%) of patients at 3 months and in 90% (95% CI 86%, 95%) at 1 year after therapy. Bulk symptoms improved in 93% of patients (95% CI 88%, 96%) at 3 months and in 91% (95% CI 86%, 95%) at 1 year after treatment. Only one major periprocedural complication occurred (pulmonary embolus), which resolved with anticoagulant therapy. Subsequent gynecologic interventions occurred in 10.5% of the patients (95% CI 7.0%, 15.0%) during the follow-up period. CONCLUSION: Uterine artery embolization is safe and controls the symptoms caused by leiomyomata in most patients. PMID- 11430953 TI - Outcomes after rollerball endometrial ablation for menorrhagia. AB - OBJECTIVE: To evaluate the outcomes of women undergoing rollerball endometrial ablation for menorrhagia and to identify factors associated with those outcomes. METHODS: Data on the clinical history, operative technique, and follow-up status as of August 1998 were obtained by retrospective medical record review for 240 women undergoing rollerball endometrial ablation with or without resection of polyps or myomas from January 1991 through December 1996. The incidence of subsequent hysterectomy was calculated by survival analysis, and Cox proportional hazard models were used to identify the predictors of success or failure of the procedure. RESULTS: The mean follow-up time was 31.2 months. Twenty-nine women (31% of the available subjects) who had not undergone hysterectomy were still being monitored 5 years after the rollerball endometrial ablation. Overall, the probability of no hysterectomy in the first 5 years was 71%. Ablation was repeated in 10 patients, six of whom eventually underwent hysterectomy. Multivariate analysis identified previous tubal ligation as a statistically significant positive predictor of the risk of hysterectomy (hazard ratio 2.20, 95% confidence interval [CI] 1.18, 4.09). Women at least 45 years old had a lower risk of subsequent hysterectomy than those younger than 35 years of age (hazard ratio 0.28, 95% CI = 0.10, 0.75). CONCLUSION: The results of this study confirm the effectiveness of rollerball endometrial ablation for the treatment of menorrhagia for a longer duration of follow-up than in most previous reports. Repeated ablation and a younger age at the time of ablation increase the risk of requiring a subsequent hysterectomy. PMID- 11430954 TI - Iliococcygeus or sacrospinous fixation for vaginal vault prolapse. AB - OBJECTIVE: To compare iliococcygeus (prespinous) and sacrospinous fixation for vaginal vault prolapse. METHODS: Between 1994 and 1998, 78 women underwent sacrospinous colpopexy and 50 underwent iliococcygeus fixation for the management of symptomatic vaginal vault prolapse. A matched case-control study was designed to compare the two approaches. The matched variables included age, parity, body mass index, degree of vault prolapse, menopause, sexual activity, constipation, previous prolapse or continence surgery, stress incontinence, and length of review. Thirty-six matched pairs were isolated, resulting in a study with a power of 50% to detect a 20% difference in the success rates between the two groups. RESULTS: The subjective success rate for the iliococcygeus group was 91%; it was 94% for the sacrospinous group (P =.73). The objective success rate was 53% and 67% (P =.36), and the patient satisfaction with surgery was 78 of 100 and 91 of 100 (P =.01) on a visual analogue scale. The mean length of postoperative follow up was 21 months for the iliococcygeus group and 19 months for the sacrospinous group (P =.52). The recovery time was 54 days in the iliococcygeus group and 39 days in the sacrospinous group (P =.04). No significant difference was seen in the incidence of postoperative cystoceles or damage to the pudendal neurovascular bundle. CONCLUSION: Sacrospinous and iliococcygeus fixation are equally effective procedures for vaginal vault prolapse and have similar rates of postoperative cystocele, buttock pain, and hemorrhage requiring transfusion. The sacrospinous ligament fixation should not be discarded in favor of the iliococcygeus fixation in the management of vaginal vault prolapse. PMID- 11430955 TI - Vulvar vestibulitis syndrome: reliability of diagnosis and evaluation of current diagnostic criteria. AB - OBJECTIVE: To assess the reliability of the diagnosis of vulvar vestibulitis as defined by Friedrich and to evaluate the usefulness of Friedrich's criteria in the diagnostic process. METHODS: In a university hospital, 146 women with dyspareunia had two sets of gynecologic examinations involving vulvar pain ratings, took part in structured interviews, and completed the McGill-Melzack Pain Questionnaire. RESULTS: Kappa values for the vulvar vestibulitis diagnosis ranged from 0.66 to 0.68 for inter-rater agreement and from 0.49 to 0.54 for test retest reliability. Mean vestibular pain ratings ranged from 2.45 at the 12 o'clock site to 7.58 at the 9-12 o'clock site; ratings for all sites correlated significantly between gynecologists. Pain in the labia majora and labia minora was minimal for both sets of examinations, with mean participant pain ratings ranging from 0 to 1.49. Gynecologists' erythema ratings did not correlate significantly with respect to either inter-rater agreement or test-retest reliability. Of Friedrich's three diagnostic criteria, only tenderness to pressure within the vulvar vestibule differentiated dyspareunia patients with and without vulvar vestibulitis. In reference to their coital pain, 88.1% of women with vulvar vestibulitis chose adjectives from the McGill-Melzack Pain Questionnaire describing a thermal quality, and 86.6% chose adjectives describing an incisive pressure sensation. CONCLUSION: Vulvar vestibulitis can be reliably diagnosed in women with dyspareunia. Pain is limited to the vulvar vestibule and can be rated and described in a consistent fashion by these women. Erythema does not appear to be a useful diagnostic criterion. PMID- 11430956 TI - Bladder neck circulation by Doppler ultrasonography in postmenopausal women with urinary stress incontinence. AB - OBJECTIVE: To investigate the effect of hormone replacement therapy (HRT) on bladder neck circulation in postmenopausal women with genuine urinary stress incontinence (USI). METHODS: A total of 227 women were enrolled in the study, including 114 postmenopausal women with USI (n = 57) or without USI (n = 57) and 113 premenopausal women with USI (n = 55) or without USI (n = 58). The bladder neck circulation was measured in 31 postmenopausal women with USI and in 12 without USI, all of whom received 0.625 mg conjugated equine estrogen plus 5 mg medroxyprogesterone acetate daily for 6 months. Bladder neck blood flow data detected by perineal color Doppler ultrasonography and weekly urinary diary data were collected before HRT and then 3 and 6 months after beginning HRT. Two-way analysis of variance and repeated measures of variance were used for statistical analysis. Power analysis was done by a two-sided test with a significance level at.05. RESULTS: The pulsatility index (PI) was significantly higher in the postmenopausal than in the premenopausal women. The postmenopausal women with USI had the highest PI level. The presence of USI did not change the PI values in the premenopausal women. After 3 months of HRT, the PI levels decreased significantly (P <.001) in the postmenopausal women with USI. The subjective improvement of USI appeared after 3 months of HRT. CONCLUSION: The blood flow around the bladder neck in women can be measured by perineal color Doppler ultrasonography. Hormone replacement therapy increases the blood flow around the bladder neck in postmenopausal women with USI. The clinical improvement of USI can be seen with HRT after 3 months. PMID- 11430957 TI - Causes and consequences of recent increases in preterm birth among twins. AB - OBJECTIVE: To examine the causes and consequences of the recent increase in preterm birth among twins. METHODS: We studied all twin births among residents of the province of Nova Scotia, Canada, between 1988 and 1997. Rates of preterm birth, preterm labor induction, preterm cesarean, small-for-gestational age (SGA), respiratory distress syndrome (RDS), stillbirth, perinatal mortality, and infant mortality were compared between past and more recent years. Changes in perinatal mortality were examined using logistic regression to adjust for the effects of other determinants. RESULTS: The study included 2516 twin births (73 stillbirths and 2443 live births). The rate of preterm birth increased from 42.3% in 1988-1992 to 48.2% of twin live births in 1993-1997 (14% increase, P =.04). Twin live births born after preterm labor induction increased from 3.5% in 1988 1989 to 8.6% in 1996-1997 (P for trend =.007). Of live births between 34 and 36 weeks' gestation, the proportion born SGA decreased from 17.5% in 1988-1992 to 9.2% in 1993-1997 (P =.005). Over the same period, rates of prophylactic maternal steroid therapy increased substantially and rates of RDS declined. Perinatal mortality rates among pregnancies reaching 34 weeks decreased from 12.9 per 1000 total births in 1988-1992 to 4.2 per 1000 total births in 1993-1997 (P =.05). CONCLUSION: Increases in preterm labor induction appear to be responsible for the recent increase in preterm birth among twins. These changes have been accompanied by decreases in perinatal morbidity and mortality among twin pregnancies that reach 34 weeks' gestation. PMID- 11430958 TI - Low 5-minute Apgar score: a population-based register study of 1 million term births. AB - OBJECTIVE: To determine the rate of 5-minute Apgar scores below 7 in term infants (at least 37 weeks) in Sweden during 1988-1997, evaluate the influence of obstetric risk factors on low 5-minute Apgar scores, and to study the infant prognosis regarding infant mortality, neonatal neurologic morbidity, and outcome. METHODS: Data were collected from the Swedish Medical Birth Registry 1988-1997, and the National Hospital Discharge Registry. Odds ratios (OR) and risk ratios were calculated. RESULTS: Among 1,028,705 term newborns, 7787 (0.76%) had 5 minute Apgar scores below 7. The annual rate of low Apgar scores decreased from 0.77% in 1988 to 0.63% in 1992, but thereafter increased to 0.82% in 1998. The highest OR was found for vaginal breech delivery (OR 6.7), birth weights above 5 kg (OR 6.3), and second born twins (OR 4.1). Primiparity, maternal age, smoking, post-date pregnancy, epidural analgesia, male infant gender, and being born at night, were also significant risk factors for Apgar below 7 at 5 minutes. The infant mortality rate was 48 per 1000 (OR 14.4), and the ORs were 31.4 for a diagnosis with cerebral palsy, 7.9 for epilepsy, and 9.5 for mental retardation. CONCLUSION: Several obstetric risk factors are associated with low 5-minute Apgar score in term infants. Mortality and the risk of severe neurologic morbidity are increased in these infants. PMID- 11430959 TI - Ultrasound estimate of amniotic fluid volume: color Doppler overdiagnosis of oligohydramnios. AB - OBJECTIVE: To determine if concurrent use of color Doppler affects ultrasound estimates of amniotic fluid (AF) volume. METHODS: Study gravidas underwent ultrasound estimations of AF volume subjectively (visualization without measurements) and objectively (using amniotic fluid index [AFI]) and single deepest pocket techniques, without and with concurrent color Doppler. Amniocentesis with the dye-dilution technique to measure actual AF volume was utilized for comparison. RESULTS: Sixty-seven women at a mean gestational age of 37.1 +/- 2.5 weeks were entered into this investigation between June 1999 and March 2000. Dye-determined AF volume was classified as low in 18 patients and as high in seven, with the remaining 42 within normal range. Using either ultrasound technique with color Doppler produced significantly lower estimates of AF volume (9.3 +/- 4.9) compared to those without color ([11.6 +/- 5], P <.001) for the AFI and (3.7 +/- 1.5) with color compared to those without color ([4.5 +/- 1.5], P <.003) for the single-deepest pocket. Using AFI without color identified two of 67 (3%) of the pregnancies as having low fluid compared to 14 of 67 ([21%] P =.002) using color. The increased classification of oligohydramnios with color did not accurately identify a greater number of dye-determined low volumes; instead, the AFI with color mislabeled nine pregnancies with normal fluid as low. The diagnosis of dye-determined low and high fluid volumes was not significantly different with or without color. CONCLUSION: Concurrent use of color Doppler with AFI measurements leads to the overdiagnosis of oligohydramnios. PMID- 11430960 TI - Umbilical cord serum ionized magnesium level and total pediatric mortality. AB - OBJECTIVE: To estimate whether higher magnesium levels in umbilical cord blood at delivery are associated with increased total pediatric (fetal + neonatal + postneonatal) mortality. METHODS: During the Magnesium and Neurologic Endpoints Trial, in addition to randomizing mothers having preterm labor into arms containing magnesium sulfate, other tocolytic agents, or saline controls, we obtained biologic specimens at delivery, including umbilical cord venous blood on which was determined the serum ionized magnesium level using the AVL 988-4 analyzer (Graz, Austria). Laboratory results were then matched with the pediatric mortalities. The study power was based on the anticipated reductions in neonatal intraventricular hemorrhage related to magnesium usage from 18.9% to 4.4%. For alpha =.05, 1-beta (power)=80%, two tailed, the total number of infants needed would be 140. RESULTS: Of 149 mothers who gave permission for randomization, ionized magnesium levels were available for 82 children. Seven deaths occurred (one immediately before delivery, three as neonates, and three in the postneonatal period). The median level of ionized magnesium among the seven dead children was 0.76 mmol/L; among the 75 survivors, the median level of ionized magnesium was 0.55 mmol/L (Mann-Whitney U test, P =.03). Using multivariable logistic regression analysis, the association remained statistically significant when controlling for possible confounding factors (adjusted odds ratio 7.7, 95% confidence interval 1.2, 47.6, P =.03). CONCLUSION: These findings of a dose response between serum ionized magnesium and deaths in children increase our concern about the improper use of tocolytic magnesium. PMID- 11430961 TI - Nugent score related to vaginal culture in pregnant women. AB - OBJECTIVE: To relate Gram-stained smears, using the Nugent criteria, to quantitative and qualitative vaginal cultures in pregnant women. METHODS: Two independent evaluators using the Nugent criteria, a standardized method of Gram stain interpretation designed to detect bacterial vaginosis, scored 104 vaginal smears from pregnant women. The quantitative and qualitative vaginal cultures were assessed at the same time and the results expressed as log(10) colony forming units per gram of vaginal secretion. The Nugent scores were compared with the microbiologic findings. RESULTS: The prevalence of normal, intermediate, or bacterial vaginosis vaginal flora as determined by Gram stain was 68%, 21%, and 11%, respectively. A comparison of the mean bacterial counts with the Nugent score showed a weak negative correlation for Lactobacillus species and a positive correlation for gram-variable and gram-negative rods. Additional analysis revealed a strong positive correlation between the mean bacterial counts analyses of Peptostreptococcus, a genus not included in the Nugent scoring system, and the Nugent score. In addition, the Prevotella counts correlated strongly with both the Nugent score and the Peptostreptococcus counts. The quantitative counts for Lactobacillus did not vary significantly among the three defined groups of vaginal microflora; however, significant increases in the concentrations of Gardnerella vaginalis and Prevotella were found as the Nugent score increased. CONCLUSION: A strong correlation was found among the gram-variable and gram negative genera comprised by the Nugent score. Peptostreptococcus also correlated strongly with the Nugent score and with the Prevotella counts, suggesting that this genus may play a role in determining vaginal health. PMID- 11430962 TI - Age at natural menopause and the risk of epithelial ovarian cancer. AB - OBJECTIVE: To investigate the relationship between age at natural menopause and risk of developing epithelial ovarian cancer. METHODS: Using data from six population-based, case-control studies conducted in the United States, age at natural menopause among 1411 women with epithelial ovarian cancer and 6380 control subjects were analyzed using survival analysis methods, including Kaplan Meier and proportional hazards models. Subjects ranged from 20 to 81 years of age. RESULTS: The median age at natural menopause was 50 years among cases compared with 51 years among controls, a difference of borderline statistical significance (P =.06). The hazard ratio for the relationship between case-control status and age at natural menopause was 1.09 (95% confidence interval 0.99, 1.20). Controlling for potential confounders including parity, oral contraceptive use, tubal ligation, smoking, and body mass index did not appreciably change this association. There was little evidence of an association between early age at natural menopause and early onset ovarian cancer (diagnosis age under 48 years). CONCLUSION: We observed a weak association between ovarian cancer risk and age at natural menopause and, among women with early onset disease, there was little evidence to suggest that early menopause is related to ovarian cancer. Thus, there seems little need for increased surveillance or screening for ovarian cancer among women with early natural menopause. PMID- 11430963 TI - Raloxifene effect on frequency of surgery for pelvic floor relaxation. AB - OBJECTIVE: To assess the effects of raloxifene therapy on the frequency of surgery for pelvic floor relaxation in postmenopausal women. METHODS: This analysis used safety data through 3 years of treatment from three double-masked, placebo-controlled, randomized trials of raloxifene, which included 6926 postmenopausal women with uteri at entry. Studies 1 and 2 enrolled 969 nonosteoporotic, postmenopausal women who were assigned to 30, 60, or 150 mg per day raloxifene or placebo. Study 3 enrolled 5957 osteoporotic, postmenopausal women randomized to raloxifene 60 or 120 mg per day or placebo. Indications for any reported pelvic operations were identified, including procedures performed for pelvic organ prolapse or urinary incontinence. RESULTS: A total of 34 (1.51%) women in the placebo group and 35 (0.75%) raloxifene-treated women underwent surgical procedures for pelvic floor relaxation. The odds ratio (and 95% confidence interval) for pelvic floor repair in women assigned to raloxifene was 0.50 (0.31, 0.81). Thus, raloxifene therapy was associated with a significantly reduced risk for pelvic floor surgery (P <.005). CONCLUSION: Raloxifene does not increase pelvic floor relaxation. An apparent protective effect on pelvic floor function warrants further investigation. PMID- 11430964 TI - Intraperitoneal gas drain to reduce pain after laparoscopy: randomized masked trial. AB - OBJECTIVE: To determine whether a drain placed in the peritoneal cavity during laparoscopy is both a clinical and cost-effective method of reducing postoperative pain. METHODS: Two hundred twenty-five women undergoing diagnostic or minor operative laparoscopic procedures were recruited. Women were assigned to receive either an intraperitoneal gas drain or a dummy drain during surgery. Sample size to detect a two-point difference in visual analogue score was estimated at 158 subjects, with 79 in each arm. The patients and nursing staff were unaware of the position of the drain. A visual analogue score was used to assess pain preoperatively and at 4, 24, and 48 hours postoperatively. Data on the experience of nausea, frequency of vomiting, and site of pain were collected. The analgesic and antiemetic use was recorded. An economic evaluation of the analgesic use and the material costs for the two groups was performed. RESULTS: One hundred sixty-one complete sets of data (72%) were available for analysis. The two groups were well matched for age, parity, previous surgery, body mass index, volume of carbon dioxide used, and operative time. No significant differences were found between the two groups with regard to the overall pain scores preoperatively (8 versus 7) or at 4 (30 versus 34), 24 (40 versus 44), and 48 (26 versus 26) hours postoperatively, after adjusting for multiple point testing. On assessment at different sites, the dummy drain group experienced shoulder pain more frequently at 4 (19 of 79 versus 10 of 82, P =.05) and 48 (16 of 79 versus 7 of 82, P =.03) hours postoperatively compared with the drain group. The placebo group had a 33% greater usage of oral analgesia after discharge, but this was $2.50 cheaper than the use of an intraperitoneal drain. No statistically significant differences were found between the groups with regard to nausea and vomiting postoperatively. CONCLUSION: An intraperitoneal drain after minor gynecologic laparoscopy decreases the frequency of shoulder pain and reduces postoperative analgesia requirements. However, it is less cost effective to reduce pain using an intraperitoneal gas drain than simple oral analgesia after minor gynecologic laparoscopy. PMID- 11430965 TI - Curettage after mifepristone-induced abortion: frequency, timing, and indications. AB - OBJECTIVE: To characterize women who underwent curettage after medical abortion with mifepristone and vaginal misoprostol, to describe when curettage occurred and the reasons for the intervention, and to categorize the indications for curettage according to a simple classification schema. METHODS: These analyses used data from two multisite, randomized controlled trials sponsored by Abortion Rights Mobilization. In the first study, women pregnant less than 63 days received 200 mg of mifepristone and 800 microg of vaginal misoprostol to use 48 hours after taking mifepristone. In the second study, women pregnant less than 56 days were randomly assigned to insert vaginal misoprostol at 1, 2, or 3 days after mifepristone administration. Case report forms and clinical case notes of all women who underwent curettage were examined. RESULTS: Of the 4393 women enrolled in both studies, 116 (2.6%, 95% confidence interval 2.1%, 3.1%) curettages were identified. The gestational age and a history of prior elective abortion were associated with the rate of curettage. The distribution of indications for curettage included bleeding, 61 (53%); continuing pregnancy, 17 (15%); patient request, 36 (31%); and indeterminate, 2 (1.7%). The timing of curettage differed by the indication and scheduled interval between study visits. The distribution of the timing was bimodal. One subset of women, 44 (38%), underwent curettage in the first study week and another subset, 43 (37%), during weeks 3-5. CONCLUSION: Curettage after medical abortion with mifepristone and vaginal misoprostol is rare. At least one half of the curettages were performed for persistent bleeding several weeks after treatment. Both acute heavy bleeding and continuing pregnancy are extremely rare. PMID- 11430966 TI - Safety and efficacy of misoprostol orally and vaginally: a randomized trial. AB - OBJECTIVE: To compare the safety and efficacy accompanying oral and vaginal misoprostol for cervical ripening. METHODS: One thousand four women with medical or obstetric indications for labor induction and unripe cervices were randomly assigned to receive oral or vaginal misoprostol. Initial doses of 200 microg oral and 50 microg vaginal misoprostol were increased to 300 microg oral and 100 microg vaginal after two doses, to a maximum of six doses. Misoprostol was given every 6 hours in both groups. We anticipated that 11% of women treated vaginally would require intervention during the ripening process. Intervention was defined as interruption of the ripening process before labor or Bishop score of 7 or a lack of response to six misoprostol doses. RESULTS: Five hundred three subjects were assigned to oral and 501 to vaginal administration. Oral misoprostol was associated with significantly higher frequencies of intervention (67 [13.3%] versus 42 [8.4%], P =.01), tachysystole (114 [23.6%] versus 85 [17.6%], P =.02), and hyperstimulation (90 [18.6%] versus 66 [13.7%], P =.04). There were no significant differences in cesarean rates (147 [29.2%] versus 120 [24.0%], P =.06), mean number of misoprostol doses used (1.5 versus 1.6, P =.18), or hours from drug administration to delivery (24.5 versus 25.4, P =.77) between the oral and vaginal groups, respectively. The numbers of deliveries between the groups within 24 hours was different (271 [56%] versus 290 [60%], P =.02), oral and vaginal, respectively. No adverse neonatal outcomes were noted. CONCLUSION: Oral misoprostol has similar efficacy as vaginal misoprostol but is associated with a higher frequency of excessive uterine contractility and intervention. PMID- 11430967 TI - Early feeding after cesarean: randomized trial. AB - OBJECTIVE: To study the rate of ileus symptoms and hospital course of women who are offered solid food shortly after cesarean delivery. METHODS: This study involved women delivered by cesarean under regional anesthesia. Exclusion criteria included general anesthesia, magnesium sulfate, intra-operative bowel injury or bowel surgery, or other conditions that precluded early feeding. Early fed women were offered regular diets within 8 hours of surgery, and controls were given nothing by mouth for 12-24 hours, advanced to clear liquids on the first postoperative day, and then given solid food on the second or third postoperative day. RESULTS: Sixty women were assigned randomly to each method. Early-fed women received solid food sooner after surgery, 5.0 +/- 1.2 hours versus 40.0 +/- 10.6 hours. The incidences of mild ileus symptoms and postoperative complications were similar in both groups; however, the study did not have an adequate sample size to definitively assess safety concerns. Women in the early-fed group had shorter hospital stays (49.5 +/- 12.7 hours versus 75.0 +/- 12.3 hours, P <.001), and shorter time intervals from surgery to bowel movement, 34.5 hours (interquartile range 25.3-48.8) versus 51.0 (43.3-62.0) hours, P <.001. In the early-fed group, women whose operative times exceeded 40 minutes were more likely to have symptoms of mild ileus. CONCLUSION: Early initiation of solid food after cesarean delivery appears to be well tolerated and may be associated with a shorter hospital stay. Early-fed women whose operations exceed 40 minutes may be more likely to have mild ileus symptoms. PMID- 11430968 TI - Effects of oxytocin receptor antagonist atosiban on pregnant myometrium in vitro. AB - OBJECTIVE: To investigate dose-dependent effects of oxytocin receptor antagonist, atosiban, on oxytocin-induced contractions of myometrial strips from healthy pregnant women. METHODS: During elective cesareans, myometrial biopsies were taken from the lower uterine segment and trimmed into 2 x 2 x 10-mm longitudinal strips within 36 hours. One hundred twenty-two myometrial strips showed regular spontaneous contractions and were used for measurement. Each myometrial strip was mounted in a physiologic organ bath, and isometric contraction activity and contractile patterns were analyzed after stimulation with oxytocin. Frequency and amplitude of contractions and area under force-time curves were determined by adding atosiban at concentrations of 1, 5, 10, 20, 50, 100, 250, and 500 microg/mL. Resulting contraction activity (at least 20 contractions per concentration) was compared with reference activity. RESULTS: Atosiban had significant inhibitory effects on myometrial contractions at concentrations as low as 1 microg/mL. Compared with reference activity (oxytocin stimulation only), resulting inhibition was higher than 50% with the lowest atosiban concentration used. This inhibition of contraction activity was dose-dependent and referred to the area under the curve, frequency, and amplitude of contractions. At high concentrations of atosiban, contraction activity was even lower than spontaneous activity without oxytocin stimulation. CONCLUSION: The oxytocin receptor antagonist atosiban showed a significant, dose-dependent inhibition of oxytocin induced contractions of human myometrium in vitro. It might be effective in tocolysis at term. PMID- 11430969 TI - Cytokine patterns in maternal blood after premature rupture of membranes. AB - OBJECTIVE: To compare two types of cytokines, type 1, which activate cell mediated reactions and are important in cytotoxic and delayed-type hypersensitivity reactions, and type 2, which encourage vigorous antibody production and are commonly found in association with humoral immune responses, in blood of women with premature rupture of membranes (PROM). METHODS: Forty-four women with histories of at least three successful pregnancies and who currently delivered normally served as controls. The PROM group consisted of 30 women with spontaneous rupture of fetal membranes at term. Peripheral blood mononuclear cells were stimulated separately with a mitogen, placental cells, and a trophoblast antigen extract, and the supernatants examined for type 1 and type 2 cytokines. RESULTS: Mitogen-stimulated blood cells produced significantly higher levels of type 1 cytokines in PROM women than in normal controls. Higher levels of the type 1 cytokine interferon-gamma were produced by PROM samples stimulated with autologous placental cells and with trophoblast antigens. Ratios of type 1 to type 2 cytokines were higher in PROM compared with normal pregnancy, and in some cases as much as 25-fold higher. CONCLUSION: Women in the PROM group had a stronger type 1 reactivity whereas normal women were more predisposed to type 2 immunity; thus, PROM appears to be associated with a maternal type 1 bias. PMID- 11430970 TI - Gynecologic presentation of interstitial cystitis as detected by intravesical potassium sensitivity. AB - OBJECTIVE: To document the initial clinical diagnoses, determine the prevalence of urinary symptoms, and test for intravesical potassium sensitivity in gynecologic patients with chronic pelvic pain. METHODS: Gynecologists at three United States medical centers administered the Potassium Sensitivity Test to consecutive unselected pelvic pain patients. Before testing, each patient was given an initial clinical diagnosis based on the patient's chief symptomatic complaint(s) and surveyed for urologic symptoms. RESULTS: Of 134 patients, 114 (85%) had positive potassium sensitivity. Positive potassium sensitivity rates were similar across all three sites and all clinical diagnoses including endometriosis, vulvodynia (vulvar vestibulitis), and pelvic pain. A total of 75% of the subjects reported urologic symptoms, but only 2.9% received an initial diagnosis of interstitial cystitis. CONCLUSION: A significant majority of gynecologic patients presenting with pelvic pain have a positive Potassium Sensitivity Test, indicating their pain may have a bladder component (interstitial cystitis). Interstitial cystitis deserves greater consideration in the differential diagnosis of chronic pelvic pain. PMID- 11430971 TI - Involving the male partner for interpreting the basal body temperature graph. AB - OBJECTIVE: To determine if the male cohabiting partner of a woman may serve as a control for exogenous influences on basal body temperature (BBT). METHODS: Twelve couples from the Atlanta area were enrolled for a total of 41 couple-cycles. Couples recorded their oral temperatures daily and used urinary test kits for luteinizing hormone to estimate the day of ovulation. The covariability between the pre-ovulatory temperature of the women and their partners was assessed. The gaps in the couples' temperatures (female temperature minus male temperature) were compared in the pre- and postovulatory phases. RESULTS: Considerable covariability was found between temperatures of partners in the pre-ovulatory phase (covariance parameter = 0.49; P <.001). The pre- and postovulatory temperature gaps for all couples were significantly different in size (P <.001). For all couple-cycles, the size of the mean postovulatory temperature gap was at least 0.3-degree Fahrenheit greater than the mean pre-ovulatory temperature gap. CONCLUSION: Recording the BBT of women's partners may improve interpretation and accuracy of the BBT method. An increase in the size of a couple's temperature gap accompanies the transition from the pre- to the postovulatory phase. By this method, a given couple could determine their unique temperature gap indicating this transition. PMID- 11430972 TI - Managed care and perinatal regionalization in Washington State. AB - OBJECTIVE: To determine if an association exists between managed care penetration and perinatal deregionalization in Washington State. METHODS: The proportions of low birth weight (LBW) and very low birth weight (VLBW) deliveries were tabulated for each hospital in Washington State for the years 1989, 1993 and 1996. Level of perinatal care, degree of health maintenance organization (HMO) penetration, and maternal demographic characteristics including age, race, smoking, and Medicaid status were derived from state and national databases. Multiple linear regression analysis was performed for each hospital level to evaluate the association between change in proportion of LBW and VLBW deliveries and change in HMO penetration per hospital between each of the 3 years. RESULTS: From 1989 through 1993, the proportion of LBW deliveries significantly declined at level III hospitals and rose at level I and II hospitals. This trend reversed between 1993 and 1996. Very low birth weight deliveries demonstrated more limited and somewhat contrary results, significantly decreasing, then increasing in level I hospitals, and significantly increasing in level III hospitals from 1989 to 1993. After controlling for changes in maternal characteristics over time, changes in HMO penetration at the hospital level were not significantly associated with an increasing proportion of LBW or VLBW deliveries at nonlevel III hospitals. In some analyses, increasing HMO penetration actually was significantly associated with decreasing LBW and VLBW deliveries at nonlevel III hospitals. CONCLUSION: Despite continued growth in HMOs throughout the state, the trend toward deregionalization in Washington State noted in the early 1990s has not continued. At the hospital level, the increasing presence of HMOs is not significantly associated with perinatal deregionalization. PMID- 11430973 TI - Antenatal corticosteroids revisited: repeat courses - National Institutes of Health Consensus Development Conference Statement, August 17-18, 2000. AB - OBJECTIVE: To provide health care providers, patients, and the general public with a responsible assessment of currently available data regarding the benefits and risks of repeat courses of antenatal corticosteroids. PARTICIPANTS: A non Federal, non-advocate, 16-member panel representing the fields of obstetrics and gynecology, pediatrics, maternal and fetal medicine, neonatology, medical ethics, community health, pharmacology, psychology, and reproductive biology. In addition, 13 experts in these same fields presented data to the panel and to a conference audience of approximately 200. EVIDENCE: The literature was searched using MEDLINE and an extensive bibliography of references was provided to the panel. Experts prepared abstracts of their conference presentations with relevant citations from the literature. Scientific evidence was given precedence over clinical anecdotal experience. CONSENSUS PROCESS: The panel, answering predefined questions, developed their conclusions based on the scientific evidence presented in open forum and the scientific literature. The panel composed a draft statement that was read in its entirety and circulated to the experts and the audience for comment. Thereafter, the panel resolved conflicting recommendations and released a revised statement at the end of the conference. The panel finalized the revisions within a few weeks after the conference. The draft statement was made available on the World Wide Web immediately following its release at the conference and was updated with the panel's final revisions. CONCLUSIONS: The collective international data continue to support unequivocally the use and efficacy of a single course of antenatal corticosteroids using the dosage and interval of administration specified in the 1994 Consensus Development Conference report. The current benefit and risk data are insufficient to support routine use of repeat or rescue courses of antenatal corticosteroids in clinical practice. Clinical trials are in progress to assess potential benefits and risks of various regimens of repeat courses. Until data establish a favorable benefit-to-risk ratio, repeat courses of antenatal corticosteroids, including rescue therapy, should be reserved for patients enrolled in clinical trials. PMID- 11430974 TI - Emergency contraception over-the-counter: the medical and legal imperatives. AB - Requiring a physician's prescription for hormonal emergency contraceptive pills makes no sense. Unintended pregnancies remain endemic in the United States, and wider use of emergency contraceptive pills could substantially help. However, the prescription requirement poses an unnecessary barrier to prompt, effective use of this preventive therapy. According to the Durham-Humphrey Amendment of 1951, the default option for all new drugs is, in principle, over-the-counter, unless a drug is addictive or dangerous when self-administered. Clearly, hormonal emergency contraception is neither of these. Emergency contraceptive pills meet all the customary criteria for over-the-counter use: low toxicity, no potential for overdose or addiction, no teratogenicity, no need for medical screening, self identification of the need, uniform dosage, and no important drug interactions. The Food and Drug Administration is authorized, and, by its own regulations, should be required to switch hormonal emergency contraception to over-the-counter status without delay. The current prescription requirement is not only gratuitous but also harmful to women's health because it impedes access to this important therapy. PMID- 11430975 TI - Professionalism: a key to weathering the storm. AB - The President's Program at the 2000 Annual Clinical Meeting of ACOG addressed the question of how best to maintain professional values in the current, market driven health care environment. The author's presentation during that presidential program provided the basis of this article. In today's corporate environment, the distinction between the practice of medicine and the business of medicine has become blurred. Too often, market values prevail over traditional professional values. As a consequence, physicians face increasing challenges and frustrations. To ease the discomfort of practicing in a new corporate age, physicians must maintain medical professionalism and hence reassert the primacy of professional values in caring for patients. Individually, physicians must exercise professionalism in their roles as educators and practitioners. Collectively, the profession of medicine must exercise professionalism by advocating patients' interests and by accepting accountability for both long established and emerging obligations that physicians have to their patients and to society. PMID- 11430977 TI - Insight into multi-site mechanisms of glycosyl transfer in (1-->4)beta-D-glycans provided by the cereal mixed-linkage (1-->3),(1-->4)beta-D-glucan synthase. AB - Synthases of cellulose, chitin, hyaluronan, and all other polymers containing (1- >4)beta-linked glucosyl, mannosyl and xylosyl units have overcome a substrate orientation problem in catalysis because the (1-->4)beta-linkage requires that each of these sugar units be inverted nearly 180 degrees with respect to its neighbors. We and others have proposed that this problem is solved by two modes of glycosyl transfer within a single catalytic subunit to generate disaccharide units, which, when linked processively, maintain the proper orientation without rotation or re-orientation of the synthetic machinery in 3-dimensional space. A variant of the strict (1-->4)beta-D-linkage structure is the mixed-linkage (1- >3),(1-->4)beta-D-glucan, a growth-specific cell wall polysaccharide found in grasses and cereals. beta-Glucan is composed primarily of cellotriosyl and cellotetraosyl units linked by single (1-->3)beta-D-linkages. In reactions in vitro at high substrate concentration, a polymer composed of almost entirely cellotriosyl and cellopentosyl units is made. These results support a model in which three modes of glycosyl transfer occur within the synthase complex instead of just two. The generation of odd numbered units demands that they are connected by (1-->3)beta-linkages and not (1-->4)beta-. In this short review of beta-glucan synthesis in maize, we show how such a model not only provides simple mechanisms of synthesis for all (1-->4)beta-D-glycans but also explains how the synthesis of callose, or strictly (1-->3)beta-D-glucans, occurs upon loss of the multiple modes of glycosyl transfer to a single one. PMID- 11430978 TI - Differential behaviour of four plant polysaccharide synthases in the presence of organic solvents. AB - The behaviour of four membrane-bound glycosyl transferases involved in cell wall polysaccharide synthesis has been studied in relation to the effects of a graded series of organic solvents on their activity and type of product formed. Relative enzyme inhibition observed for some solvents was in direct relationship to the hydrophilicity of the product. This was in the order of arabinan synthase > callose synthase> xylan synthase > beta-1,4-glucan synthase. The former two were always inhibited, the xylan synthase rather less so. However, the beta-1,4-glucan synthase showed significant increases in substrate incorporation in the presence of solvents. A graded series of primary alcohols were much more effective in enhancing activity than acetone, ethyl acetate and dimethyl formamide. In the presence of the most effective solvent, methanol, there was considerable activation of beta-1,4-glucan production. This reciprocal nature of the behaviour of the beta-1,4- and beta-1,3-glucan synthases in organic solvent is supportive of recent molecular data that the two types of glucans are catalysed by separate enzyme systems. However, the results reported here do not totally negate the proposition that either enzyme is capable of synthesising the other linkage in minor amounts in vitro. PMID- 11430979 TI - Oxidative pentose phosphate pathway and pyridine nucleotides in relation to heartwood formation in Robinia pseudoacacia L. AB - Most tree species show in the inner parts of their woody axes often a dark colored zone, the heartwood. Its formation is a genetically determined, programmed cell death which is characterized by the activation of metabolic pathways which lead to the formation of phenolic heartwood extractives. In the present paper we report on the key position of the oxidative pentose phosphate pathway (OPP) for this process. The OPP plays a crucial role in anabolic processes and is involved in the interconversion and rearrangements of sugar phosphates with the net production of NADPH. In tissues of Robinia pseudoacacia L. which are transferred to heartwood, enhanced activities of glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH) are present. A consequence of these increased enzyme activities is a shift in the pyridine nucleotide pool towards NADP+NADPH at the expense of NAD+NADH. These alterations in the metabolism and the redox status probably provide precursors and reduction equivalents being required for the synthesis of heartwood phenolics. The non heartwood forming species Acer pseudoplatanus L. shows neither a radial gradient nor seasonal changes in the amounts of pyridine nucleotides across the trunkwood. The results are discussed in connection with programmed cell death, mitochondrial activity, and heartwood formation. PMID- 11430980 TI - The biosynthesis of monolignols: a "metabolic grid", or independent pathways to guaiacyl and syringyl units? AB - Lignin is a complex polymer formed by the oxidative polymerization of hydroxycinnamyl alcohol derivatives termed monolignols. The major monolignols in dicotyledonous angiosperm lignin are monomethylated guaiacyl (G) units derived from coniferyl alcohol, and dimethylated syringyl (S) units derived from sinapyl alcohol. The biochemical pathways leading to the formation of monolignols feature successive hydroxylation and O-methylation of the aromatic ring and conversion of the side chain carboxyl to an alcohol function. The current view of the monolignol biosynthetic pathway envisages a metabolic grid leading to G and S units, through which the successive hydroxylation and O-methylation reactions may occur at different levels of side chain oxidation. The present article assesses biochemical and genetic evidence for and against such a model, including recent data on the methylation reactions of monolignol biosynthesis in alfalfa. We draw attention to portions of the currently accepted monolignol pathway that may require revision, and suggest an alternative model in which metabolic channeling allows for independent pathways to G and S lignin. PMID- 11430981 TI - Coniferyl alcohol metabolism in conifers -- I. Glucosidic turnover of cinnamyl aldehydes by UDPG: coniferyl alcohol glucosyltransferase from pine cambium. AB - UDPG: coniferyl alcohol glucosyltransferase (CAGT; EC 2.4.1.111) isolated from cambial tissues of Pinus strobus was able to convert cinnamyl aldehydes as well as dihydroconiferyl alcohol into their corresponding 4-O-beta-D-glucosides in vitro. Cinnamyl aldehydes were glucosylated with comparable efficiency to coniferyl alcohol, the physiological substrate for CAGT. Seasonal patterns of CAGT activity for aldehydes were similar to those of coniferyl alcohol. Formation of cinnamyl aldehyde and additional monolignol glucosides indicates that precursor flux and availability for lignification is likely greater than previously recognized. PMID- 11430982 TI - Coniferyl alcohol metabolism in conifers -- II. Coniferyl alcohol and dihydroconiferyl alcohol biosynthesis. AB - Coniferaldehyde and NADPH when incubated with microsomes isolated from developing xylem of Pinus strobus yielded coniferyl alcohol and dihydroconiferyl alcohol in vitro. D-(+)-Pinitol was also found to be a microsomal constituent. Endogenous E coniferyl alcohol content, quantified in dormant buds, cambium, bark and needles of Pinus resinosa and P. strobus by isotope-dilution combined gas chromatography mass spectrometry (GC/MS) using ring-(13)C(6)-coniferyl alcohol, was at a level similar to that of endogenous indol 3-ylacetic acid (IAA). Wounding (branch girdling) induced more than a 10-fold increase in content of endogenous E coniferyl alcohol in dormant non-lignifying cambium, a clear indication that monolignol biosynthesis is not coupled to lignification. PMID- 11430983 TI - Oxidation of cinnamyl alcohols and aldehydes by a basic peroxidase from lignifying Zinnia elegans hypocotyls. AB - The xylem of 26-day old Zinnia elegans hypocotyls synthesizes lignins derived from coniferyl alcohol and sinapyl alcohol with a G/S ratio of 43/57 in the aryl glycerol-beta-aryl ether core, as revealed by thioacidolysis. Thioacidolysis of Z. elegans lignins also reveals the presence of coniferyl aldehyde end groups linked by beta-0-4 bonds. Both coniferyl and sinapyl alcohols, as well as coniferyl and sinapyl aldehyde, are substrates of a xylem cell wall-located strongly basic peroxidase, which is capable of oxidizing them in the absence and in the presence of hydrogen peroxide. This peroxidase shows a particular affinity for cinnamyl aldehydes with kappa(M) values in the mu(M) range, and some specificity for syringyl-type phenols. The affinity of this strongly basic peroxidase for cinnamyl alcohols and aldehydes is similar to that shown by the preceding enzymes in the lignin biosynthetic pathway (microsomal 5-hydroxylases and cinnamyl alcohol dehydrogenase), which also use cinnamyl alcohols and aldehydes as substrates, indicating that the one-way highway of construction of the lignin macromolecule has no metabolic "potholes" in which the lignin building blocks might accumulate. This fact suggests a high degree of metabolic plasticity for this basic peroxidase, which has been widely conserved during the evolution of vascular plants, making it one of the driving forces in the evolution of plant lignin heterogeneity. PMID- 11430984 TI - The poly(phenolic) domain of potato suberin: a non-lignin cell wall bio-polymer. AB - Suberized plant cell walls have three distinguishing features: (1) tissue specificity, (2) a poly(aliphatic) domain and (3) a unique, "lignin-like" poly(phenolic) domain. With respect to the latter, comparisons have often been made to lignin, but the unique phenolic composition of suberized cells yields a unique polymer better designated as a poly(phenolic) domain. Potato tubers that have been induced to suberize through wounding make an excellent model system with which the chemistry, biochemistry and macromolecular assembly of the suberin poly(phenolic) domain can be monitored. For example, wound healing potato tubers have been used to determine the unique hydroxycinnamic acid nature of its poly(phenolic) domain using specific carbon-13 labeling studies and specific chemical degradation techniques (e.g. thioacidolysis). Furthermore, a suberization-associated anionic peroxidase has been purified from suberizing potato tubers and subsequently shown to oxidize hydroxycinnamic acids (and their derivatives) in preference to monolignols, as well as yield an unique polymer in vitro. We have since extended these studies to begin analyzing the macromolecular assembly process leading to the deposition of this suberized tissue specific domain. To this end we have begun to describe an H(2)O(2)-generating system with NAD(P)H-dependent oxidase-like properties that is temporally associated with the formation of potato suberin poly(phenolics) during suberization. Herein we describe our progress to date. PMID- 11430985 TI - Similarities and differences in rubber biochemistry among plant species. AB - This report reviews aspects of the biochemical regulation of rubber yield and rubber quality in three contrasting rubber-producing species, Hevea brasiliensis, Parthenium argentatum and Ficus elastica. Although many similarities are revealed, considerable differences also exist in enzymatic mechanisms regulating biosynthetic rate and the molecular weight of the rubber biopolymers produced. In all three species, rubber molecule initiation, biosynthetic rate and molecular weight, in vitro, are dependent upon substrate concentration and the ratio of isopentenyl pyrophosphate (IPP, the elongation substrate, or monomer) and farnesyl pyrophosphate (FPP, an initiator), but these parameters are affected by intrinsic properties of the rubber transferases as well. All three rubber transferases are capable of producing a wide range of rubber molecular weight, depending upon substrate concentration, clearly demonstrating that the transferases are not the prime determinants of product size in vivo. However, despite these commonalities, considerable differences exist between the species with respect to cosubstrate effects, binding constants, effective concentration ranges, and the role of negative cooperativity in vitro. The P. argentatum rubber transferase appears to exert more control over the molecular weight it produces than the other two species and may, therefore, provide the best prospect for the source of genes for transformation of annual crop species. The kinetic data, from the three contrasting rubber-producing species, also were used to develop a model of the rubber transferase active site in which, in addition to separate IPP and allylic-PP binding sites, there exists a hydrophobic region that interacts with the linear portion of allylic-PP initiator proximal to the pyrophosphate. Substrate affinity increases until the active site is traversed and the rubber interior of the rubber particle is reached. The kinetic data suggest that the hydrophobic region in H. brasiliensis and F. elastica is about 1.8 nm long but only 1.3 nm in P. argentatum. The estimates are supported by measurements of the rubber particle monolayer membrane using electron paramagnetic resonance spectroscopy. PMID- 11430986 TI - Structure--function characterization of cellulose synthase: relationship to other glycosyltransferases. AB - A combined structural and functional model of the catalytic region of cellulose synthase is presented as a prototype for the action of processive beta glycosyltransferases and other glycosyltransferases. A 285 amino acid segment of the Acetobacter xylinum cellulose synthase containing all the conserved residues in the globular region was subjected to protein modeling using the genetic algorithm. This region folds into a single large domain with a topology exhibiting a mixed alpha/beta structure. The predicted structure serves as a topological outline for the structure of this processive beta glycosyltransferase. By incorporating new site-directed mutagenesis data and comparative analysis of the conserved aspartic acid residues and the QXXRW motif we deduce a number of functional implications based on the structure. This includes location of the UDP--glucose substrate-binding cavity, suggestions for the catalytic processing including positions of conserved and catalytic residues, secondary structure arrangement and domain organization. Comparisons to cellulose synthases from higher plants (genetic algorithm based model for cotton CelA1), data from neural network predictions (PHD), and to the recently experimentally determined structures of the non-processive SpsA and beta 4-galactosyltransferase retest and further validate our structure-function description of this glycosyltransferase. PMID- 11430987 TI - Transcriptional control of lignin biosynthesis by tobacco LIM protein. AB - Lignin is a complex phenolic plant polymer that is essential for mechanical support, defense, and water transport in higher plants. The AC-rich motif, Pal box is an important cis-acting element for gene expression in phenylpropanoid biosynthesis. We isolated a cDNA clone (Ntlim1) encoding a Pal-box binding protein by Southwestern screening. The deduced amino acid sequence of Ntlim1 is highly similar to members of the LIM protein family that contain a zinc finger motif. Moreover, Ntlim1 had a specific DNA-binding ability and transiently activated transcription of a beta-glucuronidase reporter gene driven by the Pal box sequence. The results of transient expression assays with tobacco cultured cells showed that fusion proteins between GFP and Ntlim1 can enter nuclei. Transgenic tobacco plants with antisense Ntlim1 showed low levels of transcripts from some key phenylpropanoid pathway genes such as phenylalanine ammonia-lyase, hydroxycinnamate CoA ligase and cinnamyl alcohol dehydrogenase. Furthermore, a greater than 20% reduction in lignin content was observed in transgenic tobacco with antisense Ntlim1. PMID- 11430988 TI - Antisense and sense expression of cDNA coding for CYP73A15, a class II cinnamate 4-hydroxylase, leads to a delayed and reduced production of lignin in tobacco. AB - A number of plant species contain the class II of genes encoding the cytochrome P450, CYP73, the cognate protein of which cinnamic acid 4-hydroxylase, is the second enzyme of the phenylpropanoid pathway. In order to begin to determine possible functionality, tobacco has been transformed with a truncated French bean class II cinnamate hydroxylase (CYP73A15) in the sense and antisense orientations. Signals for C4H protein could be detected in vascular tissue from wild-type plants using heterologous probes. The transformed plants showed a normal phenotype, even though detectable C4H protein was much reduced in tissue prints. Young propagated transformants displayed a range of reduced C4H activities, as well as either reduced or no phloroglucinol-stainable lignin. However, all mature tobacco plants showed the accumulation of lignin, even though its deposition was apparently delayed. This was not due to induction of tyrosine ammonia-lyase activity, which was not detected, but instead it is presumed due to sufficient C4H residual activity. Analysis of the lignin content of the plants showed reductions of up to 30% with a slightly reduced syringyl to guaiacyl ratio as compared to wild type. This reduction level was favourable in comparison with some other targets in the lignification pathway that have been manipulated including that of class I cinnamate 4-hydroxylase. It is proposed that the class II cinnamate 4-hydroxylase might also function in lignification in a number of species including French bean and tobacco, based on these data. PMID- 11430989 TI - Repression of O-methyltransferase genes in transgenic tobacco affects lignin synthesis and plant growth. AB - Among the different enzymatic steps leading to lignin biosynthesis, two methylation reactions introduce the methyl groups borne by guaiacyl (G) and syringyl (S) units. Tobacco possesses a complex system of methylation comprising three classes of CCoAOMTs (caffeoyl-CoA-O-methyltransferases) and two classes of COMTs (caffeic acid OMTs). Antisense plants transformed with the CCoAOMT sequence alone or fused to COMT I sequence have been produced and compared to ASCOMT I plants in order to study the specific role of each OMT isoform in lignin biosynthesis, plant development and resistance to pathogens. Tobacco plants strongly inhibited in OMT activities have been selected and analyzed. Whereas antisense COMT I plants exhibited no visual phenotype, CCoAOMT repression was shown to strongly affect the development of both single and double transformants: a reduction of plant growth and the alteration of flower development were observed in the most inhibited plants. Lignin analysis performed by Klason and thioacidolysis methods, showed a decrease in the lignin quantity and changes in the lignin structure of ASCCoAOMT and ASCCoAOMT/ASCOMT I transgenics but not in ASCOMT I plants. Inhibition of COMT I in single as well as in double transformed tobacco was demonstrated to decrease S unit synthesis and to provoke the accumulation of 5-hydroxyguaiacyl lignin units. ASCCoAOMT/ASCOMT I tobacco was affected in lignin amount and composition, thus demonstrating additive effects of inhibition of both enzymes. The changes of lignin profiles and the phenotypical and molecular alterations observed in the different transgenic lines were particularly prominent at the later stages of plant development. PMID- 11430990 TI - Caffeoyl coenzyme A O-methyltransferase and lignin biosynthesis. AB - Lignin, a complex phenylpropanoid compound, is polymerized from the monolignols p coumaryl alcohol, coniferyl alcohol and sinapyl alcohol. These three monolignols differ only by the 3- and 5-methoxyl groups. Therefore, enzymatic reactions controlling the methylations of the 3- and 5-hydroxyls of monolignol precursors are critical to determine the lignin composition. Recent biochemical and transgenic studies have indicated that the methylation pathways in monolignol biosynthesis are much more complicated than we have previously envisioned. It has been demonstrated that caffeoyl CoA O-methyltransferase plays an essential role in the synthesis of guaiacyl lignin units as well as in the supply of substrates for the synthesis of syringyl lignin units. Caffeic acid O-methyltransferase has been found to essentially control the biosynthesis of syringyl lignin units. These new findings have greatly enriched our knowledge on the methylation pathways in monolignol biosynthesis. PMID- 11430991 TI - Two cinnamoyl-CoA reductase (CCR) genes from Arabidopsis thaliana are differentially expressed during development and in response to infection with pathogenic bacteria. AB - Cinnamoyl-CoA reductase (CCR; EC 1.2.1.44) catalyses the conversion of cinnamoyl CoAs into their corresponding cinnamaldehydes, i.e. the first step of the phenylpropanoid pathway specifically dedicated to the monolignol biosynthetic branch. In previous work, we described the isolation and characterisation of the first cDNA encoding CCR in Eucalyptus (Lacombe, E., Hawkins, S., Van Dorsselaere, J., Piquemal, J., Goffner, D., Poeydomenge, O., Boudet, A.M., Grima-Pettenati, J., 1997. Cinnamoyl CoA reductase, the first committed enzyme of the lignin branch biosynthetic pathway: cloning, expression and phylogenetic relationships. Plant Journal 11, 429--441) and shown the role of this enzyme in controlling the carbon flux into lignins (Piquemal, J., Lapierre, C., Myton, K., O'Connell, A., Schuch, W., Grima-Pettenati, J., Boudet, A.M., 1998. Down-regulation of cinnamoyl CoA reductase induces significant changes of lignin profiles in transgenic tobacco plants. Plant Journal 13, 71--83). Here, we report the characterisation of two functionally and structurally distinct cDNA clones, AtCCR1 and AtCCR2 (81.6% protein sequence identity) in Arabidopsis thaliana. The two recombinant proteins expressed in Escherichia coli are able to use the three cinnamoyl-CoAs tested but with different levels of efficiency. AtCCR1 is five times more efficient with feruloyl-CoA and sinapoyl-CoA than AtCCR2. In addition, the two genes are differentially expressed during development and in response to infection. AtCCR1 is preferentially expressed in tissues undergoing lignification. In contrast, AtCCR2, which is poorly expressed during development, is strongly and transiently induced during the incompatible interaction with Xanthomonas campestris pv. campestris leading to a hypersensitive response. Altogether, these data suggest that AtCCR1 is involved in constitutive lignification whereas AtCCR2 is involved in the biosynthesis of phenolics whose accumulation may lead to resistance. PMID- 11430992 TI - Regulation of sex hormone-binding globulin secretion in human hepatoma G2 cells. AB - Our purpose was to examine the roles of natural (estradiol (E2) and estrone (E1)) and synthetic estrogens (ethinyl estradiol (EE), moxestrol (MOX), and tamoxifene (TAM)) in regulating production of sex hormone-binding globulin (SHBG) by human hepatoma G2 (Hep G2) cells, the rationale being that synthetic estrogens are less rapidly metabolized than natural estrogens and, thus, may alter SHBG levels more readily. In Hep G2 cells, E2, E1, and EE at 10(-7) M did not result in significantly greater SHBG secretion compared to control cells. The synthetic estrogens, MOX and TAM, caused significant, P < 0.001, increases of 30% and 51% in SHBG secretion at 10(-7) M compared to controls. However, when TAM and E2 were added together, each at 10(-7) M, no increase in SHBG secretion was noted. We conclude that natural estrogens at physiologic concentrations do not increase SHBG secretion by Hep G2 cells, but the increase of SHBG secretion caused by MOX and TAM suggests that the lack of effect of E2 and E1 may, in part, be due to their rapid metabolism. In addition, TAM stimulates SHBG secretion by interaction with the genome that is different, in certain respects, from that of E2. PMID- 11430993 TI - Development of a sensitive and specific new plasma 4-androstene-3,17- dione time resolved fluoroimmunoassay (TR-FIA). AB - We describe, for the first time to our knowledge, the development of a new, non isotopic time resolved-fluoroimmunoassay of 4-androstene-3,17-dione in plasma or serum. This steroid exhibits a key role in steroid metabolism and is often assayed in the investigation of various pathologic endocrine states. Most of the 4-androstene-3,17-dione immunoassays are performed using a radioactive tracer. We synthesized a biotinylated 4-androstene-3,17-dione tracer from 4-androstene-3,17 dione-3-carboxymethyloxime by acylation of biotinylaminopropylammonium trifluoroacetate. A specific rabbit anti 6-hemisuccinate-4-androstene-3,17 dione/BSA was indirectly bound via an anti-rabbit sheep antibody immobilized on microtiter plate wells. The amount of biotinylated-4-androstene-3,17-dione tracer was then measured by adding streptavidin-europium, and the europium fluorescence was quantified by time resolved-fluorescence (TR-FIA, Delfia System). The plasma 4-androstene-3,17-dione-levels measured with this non-isotopic assay were compared to those measured with a radioimmunoassay previously published. In both cases, the same anti-4-androstene-3,17-dione antibody was used, and the assays were performed after an extraction step and a chromatographic step. The results obtained by the two methods were virtually the same. However, the main advantages of the new plasma 4-androstenedione-3,17-dione time-resolved-fluorescence immunoassay were its greater sensitivity than radioimmunoassay and its higher precision. PMID- 11430994 TI - Selective synthesis of 4-methoxyestrogen from 4-hydroxyestrogen. AB - The introduction of an oxygen atom into the C-6 position of 4-hydroxyestrogen allowed for the selective methylation of the two phenolic hydroxyl groups. When the 6-oxo derivative of 4-hydroxyestrone was benzylated in ethanol, only the 3 monobenzyl ether was obtained without formation of the 4-monobenzyl ether. Moreover, the 6-carbonyl group was further reduced to methylene almost quantitatively in the reaction of 4-acetoxy-6-oxoestrone 3-benzyl ether derivative with sodium borohydride. Therefore, 4-methoxyestrogen was synthesized by essentially combining these two reactions. PMID- 11430995 TI - Neighboring group participation. Part 14. The preparation of the four stereoisomers of 16-hydroxymethyl-5alpha-androstane-3beta,17-diol. AB - 16alpha-Hydroxymethyl-5alpha-androstane-3beta,17beta-diol and 16beta hydroxymethyl-5alpha-androstane-3beta,17beta-diol, were obtained from reduction of 16-acetoxymethylene-5alpha-androstan-17-one. The corresponding 16alpha,17alpha and 16beta,17alpha-hydroxymethyl isomers were obtained by neighboring group participation of the 16- and 17-acetates, respectively. The reactions involving carbocation formation also led to ring D rearrangement products. PMID- 11430996 TI - Effects of internal biliary bypass on the regulation of hepatic cholesterol 7alpha-hydroxylase activity in rats. AB - In order to re-evaluate the importance of the enterohepatic circulation of bile acid in the regulation of the activities of hepatic cholesterol 7alpha hydroxylase, bile acid metabolism was examined in internal biliary bypass models of rats. A polyethylene tube was inserted into the common bile duct and another side of the tube was placed in the duodenum (DD), lower jejunum (JD), cecum (CeD), or transverse colon (CoD) as internal biliary bypass models and in the urinary bladder as an external biliary drainage (ED). After bile diversion for 7 days in each group, hepatic cholesterol 7alpha-hydroxylase activities, bile acid concentrations in bile, serum, and portal vein, biliary bile acid compositions, and intestinal absorption rates of infused labeled taurocholic acid were analyzed. Hepatic cholesterol 7alpha-hydroxylase activity was similar in the JD group compared with the DD group, however, it was significantly up-regulated in the CeD (227% of the DD group), CoD (312%), and ED groups (316%). Biliary, serum, and portal bile acid concentrations were not significantly changed in the DD, JD, and CeD groups but those were significantly lower in the CoD and ED groups compared with the DD group. The proportion of the secondary bile acids was significantly increased in the CeD group and was decreased in the CoD and ED groups. The absorption rate of taurocholic acid was almost 100%, 56%, and 23% in the JD, CeD, and the CoD group, respectively. As the cholesterol 7alpha hydroxylase activity was not significantly changed in the JD group and the predominance of secondary bile acids did not suppress the enzyme activity in the CeD group, the luminal factor, which is absorbed in the presence of bile acids, and the bile acid metabolites are not likely the regulatory factor. The cholesterol 7alpha-hydroxylase activity seems to be primarily regulated by the intestinal absorption of bile acids and partly by the intestinal mucosal factor which is linked to the intestinal bile acid absorption. PMID- 11430997 TI - New D-modified androstane derivatives as aromatase inhibitors. AB - Starting from a 16-oximino derivative of 5-androstene the newly-synthesized 16 oximino-17-hydroxy-17-substituted derivatives 2-4 gave by the Beckmann fragmentation reaction the corresponding D-seco derivatives 6-9. Besides, in the case of the 17-hydroxy-17-methyl-16-oximino derivative 2, as a result of the rearrangement, the hydrolysis product 5 of the 16-oximino group with the opposite configuration at the C-17 was obtained. By the Oppenauer oxidation and/or by dehydration of 7 with 2,3-dichloro-5,6-dicyanobenzoquinone (DDQ), the corresponding derivatives 12, 13, and 14 were obtained. The structures of 6 and 12 were unambiguously proved by the appropriate X-ray structural analysis. Kinetic analysis for anti-aromatase activity showed that compound 12 expressed the highest inhibition in the denucleated rat ovarian fraction in comparison to other androstene derivatives (IC(50) was 0.42 microM). In comparison to aminoglutethimide (AG) activity, it was 3.5 times lower. The inhibition was competitive, with K(i) of 0.27 microM. Introduction of additional units of unsaturation (compounds 13 and 14) in D-seco derivatives did not increase anti aromatase activity. PMID- 11430998 TI - Synthesis of (5alpha)-17-azaandrostan-3-ols and (5alpha)-17-aza-D-homoandrostan-3 ols and their N-acylated derivatives. AB - Two groups of N-acylated D-azasteroids (4 and 5) were synthesized to explore structure-activity relationships for steroid modulation of GABA(A) receptor function. The target compounds were prepared conveniently from (5alpha)-3 hydroxyandrostan-17-ones (6 and 7) via the intermediate (5alpha)-17-aza-D homoandrostan-3-ols (14 and 15) or (5alpha)-17-azaandrostan-3-ols (18 and 19) precursors in high overall yields. A Beckmann rearrangement and a Hofmann rearrangement were employed as two key steps in the synthetic sequences. PMID- 11430999 TI - Mechanism of action of glucocorticosteroid hormones: possible implications for therapy of neuroimmunological disorders. AB - Glucocorticosteroids are the most potent immunosuppressive and antiinflammatory drugs. Over the six decades that have passed since their discovery, a variety of genomic effector mechanisms of steroid hormones has been described which are mediated by the cytosolic steroid receptor. Recent evidence supports a direct effect of glucocorticosteroids on cellular membranes that occurs at higher hormone concentrations, termed nongenomic effects. These imply a qualitatively distinct mode of steroid action leading to cellular apoptosis. In this review, we discuss in vitro and in vivo data on nongenomic effects of glucocorticosteroids and their possible implications for the therapy of human neuroimmunological diseases. PMID- 11431000 TI - Narcolepsy and the HLA region. AB - Narcolepsy was first shown to be tightly associated with HLA-DR2 and DQ1 in 1983, suggesting a possible autoimmune mechanism. Early investigations failed to demonstrate this hypothesis, postulating that HLA-DR2 was only a linkage marker for another, unknown narcolepsy-causing gene. The autoimmune hypothesis is now being re-evaluated under the light of recent results. Like many other autoimmune disorders, narcolepsy usually starts during adolescence, is human leukocyte antigen (HLA)-associated, multigenic and environmentally influenced. Furthermore, HLA-association studies indicated a primary HLA-DQ effect with complex HLA class II allele interactions and a partial contribution of HLA to overall genetic susceptibility. Finally, recent result suggests that human narcolepsy is associated with the destruction of a small number of hypothalamic neurons containing the peptide hypocretins (orexins). This data is consistent with an immune destruction of hypocretin-containing cells as the most common etiology for human narcolepsy. PMID- 11431001 TI - Genetic linkage analysis of the antibody responses to myelin basic protein and myelin oligodendrocyte glycoprotein in rats immunized with rat spinal cord homogenate. AB - The genetic control of the antibody response to myelin basic protein (MBP) and myelin oligodendrocyte glycoprotein (MOG) was analysed in F1 and F2 crosses of DA and E3 rats, immunized with rat spinal chord homogenate. The DA rats were highly susceptible to encephalomyelitis and made antibody responses to both MBP and MOG, whereas the E3 rats were disease-resistant and responded only to MOG. The anti MBP response was mainly controlled by the disease-promoting MHC region of the DA strain together with several disease loci outside MHC. In contrast, the anti-MOG response was associated with loci not related to or actually conferring resistance to disease. PMID- 11431002 TI - Exogenous antigen containing perivascular phagocytes induce a non encephalitogenic extravasation of primed lymphocytes. AB - Recent evidence suggests that T-lymphocyte extravasation and CNS-parenchymal infiltration during autoimmune disease might be regulated by antigen-presenting (ED2(+)) cerebral/spinal perivascular phagocytes (CPP/SPP). Since the massive erythrocytic and leukocytic infiltrates in the CNS of rats with experimental allergic encephalomyelitis do not allow a precise differentiation between CPP/SPP and the invading cells in the Virchow-Robin space, we developed a new immune response model whereby the extravasation of T-lymphocytes was not followed by other blood cells. Adult Lewis rats were sensitized to horseradish peroxidase (HRP). Subsequent intracerebroventricular (i.c.v.) injections of HRP and/or Fluoro-Emerald (FE) served to: (1) challenge the primed T-lymphocytes and (2) label the CPP/SPP for additional immunocytochemical analysis. We found that 24 h and 3 days after single, double, or triple antigen boosting T-lymphocytes (R73(+), W3/25(+), OX50(+)) entered the Virchow-Robin space but did not break through the astrocytic glia limitans. Instead they adhered to HRP-containing activated CPP/SPP (mabs OX-6(+), SILK6(+), CD40(+), CD80(+), CD86(+)). This selective contact was mediated neither by cell adhesion molecules (P-selectin, ICAM-1, VCAM-1), nor promoted by chemokine receptors (CCR1, CCR5) or chemokines (monocyte chemoattractant protein (MCP)-1, MIP-1alpha, MIP-1beta, RANTES). This non-inflammatory, but antigen-dependent lymphocyte extravasation provides optimal conditions to further study the CNS immune response. PMID- 11431003 TI - Interleukin-6 increases the survival of retinal ganglion cells in vitro. AB - Interleukin-6 is a pleiotropic cytokine that mediates cellular communication both in physiological and pathological states. In this work, we demonstrate that 50 ng/mL IL-6 increases the survival of retinal ganglion cells (RGCs) after 48 h in culture. This effect was blocked by an intracellular Ca(+2) chelator, by inhibition of ryanodinic receptors and by an inhibitor of L-type Ca(+2) channels. IL-6 effect is mediated by PKC, tyrosine kinase, PI3-kinase and MEK activity. The blockade of polypeptide release also abolished the effect of IL-6. These results suggest a role for this cytokine during the development of the central nervous system (CNS). PMID- 11431004 TI - Melatonin modulation of lymphocyte proliferation and Th1/Th2 cytokine expression. AB - Melatonin is hypothesized to play a role in neuroimmunomodulation. This study investigated the in vitro effects of melatonin (10(-12) - 10(-6) M) on human peripheral blood mononuclear cell (PBMC) proliferation and T helper type 1 and T helper type 2 (Th1/Th2) cytokine expression. In vitro doses of melatonin significantly increased PBMC proliferation (p<0.05) and decreased IL-10 production in culture supernatants (p<0.05). However, there was no effect of melatonin on the stimulated production of IFN-gamma or on the intracellular accumulation of the activation antigen CD69, IFN-gamma, or IL-10 as measured by flow cytometry. These data support the notion that physiologic doses of melatonin increase lymphocyte proliferation possibly due to decreases in production of the inhibitory cytokine IL-10. PMID- 11431005 TI - Thymic epithelial cell line expresses transcripts encoding alpha-3, alpha-5 and beta-4 subunits of acetylcholine receptors, responds to cholinergic agents and expresses choline acetyl transferase. An in vitro system to investigate thymic cholinergic mechanisms. AB - Transcriptional and immunocytological characterization of thymic epithelial (TE) cell line TE750 shows that these cells, like primary TE cell cultures, transcribe alpha-3, alpha-5 and beta-4 acetylcholine receptor (AcChR) subunit genes while expressing cortical, medullary and epithelial differentiation thymic markers. Incubation of TE750 cells with nicotine decreases cell adherence and growth as measured through direct cytological observation and nucleic acid quantification, respectively. Physostigmine, a traditional cholinesterase inhibitor that also activates nicotinic AcChRs, reproduces the effects of nicotine. Strengthening the hypothesis that cholinergic receptors mediate the effects of physostigmine, acetylcholinesterase (AcChase) activity is not detected in TE750 cells. Also, like thymocytes, TE750 cells express choline acetyltransferase (ChAT), indicating that the natural transmitter AcCh can be produced locally within the thymic parenchyma. Taken together these findings indicate that TE750 cells in culture represent a suitable in vitro system for the analysis of cholinergic mechanisms operational in the thymic epithelium. PMID- 11431006 TI - Identification of delta- and mu-type opioid receptors on human and murine dendritic cells. AB - The purpose of this study was to evaluate mu- and delta-opioid receptors (OR) on human and murine dendritic cells (DC). Expression of mu- and delta-OR mRNA on DC was demonstrated by RT-PCR. The immunocytochemical and Western blot analyses revealed the expression of OR protein in DC. Radioreceptor assay demonstrated the specific saturated temperature-dependent binding of [3H]-labeled opioid ligand on DC and B(max)=2.8+/-0.3 fmol/10(6) cells and K(D)=4.8+/-1.0 nM were calculated by a Scatchard analysis. Finally, OR ligands DADLE and DAGO dose-dependently modulated the capacity of DC to induce T cell proliferation in an MLR assay. Importantly, expression of functional OR on DC was significantly increased upon TNF-alpha-induced DC maturation. Thus, these data suggest a new mechanism of opioid-dependent neuroendocrine immunomodulation. PMID- 11431007 TI - Role of poly(ADP-ribose) synthetase activation in the development of experimental allergic encephalomyelitis. AB - Peroxynitrite formation has been demonstrated during experimental allergic encephalomyelitis (EAE). Furthermore, peroxynitrite has been identified as an activator of poly(ADP-ribose) synthetase (PARS), an enzyme implicated in neurotoxicity. In the current study, we examined the role of PARS activation in the development of EAE. Administration of the PARS inhibitor 5-iodo-6-amino-1,2 benzopyrone (INH2BP) delayed the onset of EAE and reduced the incidence and severity of disease signs. Moreover, drug treatment lowered iNOS activity and decreased cell infiltration in cervical spinal tissues from EAE-sensitized animals. To conclude, the results of the present investigation suggest that PARS activity may contribute to the pathogenesis of EAE. PMID- 11431008 TI - Regulation of interleukin-6 gene expression in brain of aged mice by nuclear factor kappaB. AB - Interleukin-6 (IL-6) is increased in brain of aged mice. The purpose of this study was to determine if binding of nuclear factor kappaB (NFkappaB) to the IL-6 promoter is responsible for the age-related increase in brain IL-6. In an initial study, the effect of age on IL-6 in brain was verified as IL-6 protein was increased in brain of aged mice compared to adult and juvenile mice. Competitive RT-PCR showed that IL-6 mRNA concentration was at least 4-fold higher in aged brain compared to adult brain. Next, binding of the transcription factor NFkappaB to the IL-6 promoter in brains of 1-, 6-, and 24-month-old mice was determined. Electrophoretic mobility shift assay showed that NFkappaB activity was increased in aged brain compared to adult and juvenile brain. Moreover, glial cells cultured from aged mice showed more NFkappaB DNA-binding activity and more IL-6 mRNA and protein expression than glia from adults. However, incubating glia from aged mice in the presence of kappaB decoy inhibited these effects of age. The same was observed in vivo as intracerebroventricular injection of kappaB decoy in aged mice decreased NFkappaB activity and IL-6 mRNA and protein in brain. These results show that the DNA-binding activity of NFkappaB is increased in the brain of aged mice and that at least one consequence is increased expression of IL-6. PMID- 11431009 TI - Glutamate is a mediator of neurotoxicity in secretions of activated HIV-1 infected macrophages. AB - We sought to identify neurotoxin(s) secreted by HIV-1-infected mononuclear phagocytes that could contribute to the pathophysiology of HIV-1-associated dementia (HAD). Neurotoxic factors were characterized in batches of conditioned media (CM) from human monocyte-derived macrophages (MDM) infected with HIV-1(ADA) and/or activated with lipopolysaccharide (LPS). All of the neurotoxicity was: present in the <3000-Da fraction; blocked by 5 microM MK801; and not trypsin sensitive or extractable into polar organic solvents. Glutamate measured in CM accounted for all neurotoxic effects observed from HIV/LPS CM in astrocyte-poor neuronal cultures and may contribute to the pathophysiology of HIV-1-associated dementia. PMID- 11431010 TI - Dendritic cells presenting pyruvate kinase M1/M2 isozyme peptide can induce experimental allergic myositis in BALB/c mice. AB - Polymyositis (PM) is an inflammatory muscle disease caused by autoimmune dysfunction, considered to be caused by cytotoxic CD8 T cells. To date, no autoantigens have been identified. We attempted to induce an experimental allergic myositis (EAM) in BALB/c mice by inoculating syngeneic dendritic cells (DC) presenting peptides that are expected to match the binding anchor motif of H 2K(d) (BALB/c). We selected peptides that are highly expressed in skeletal muscle. Only when we inoculated syngeneic bone marrow-derived DC presenting pyruvate kinase M1/M2 peptide 464-472 in BALB/c mice, 41.7% of the mice (EAM) developed pathological changes in skeletal muscle compatible to human PM. Under other conditions (when we inoculated DC presenting no synthetic peptides into BALB/c or C57BL/6 mice and DC presenting pyruvate kinase M1/M2 peptide into C57BL/6 mice), there were no necrotizing and inflammatory lesions. Induction of EAM in the same manner as above also induced CTL activity against P815 cells with the same peptide and syngeneic differentiated cultured myotubes without peptides by the chromium release assay. Consistent with the similarity of the binding anchor motif of H-2K(d) (BALB/c) and HLA A*2402, we conclude that pyruvate kinase M1/M2 peptide is a candidate autoantigen not only in BALB/c-EAM but also in human PM with the HLA A*2402 allele. PMID- 11431011 TI - Comparative characterization in the rat of the interaction between cannabinoids and opiates for their immunosuppressive and analgesic effects. AB - In the present work, we investigated in the rat the possibility of functional interaction between opiate and cannabinoid systems at immune level comparatively with the central nervous system (CNS). Moderate analgesic doses of the synthetic cannabinoid compound CP-55,940 (0.2 mg/kg, i.p.) and morphine (5 mg/kg, s.c.) significantly inhibited the ConA-induced splenocyte proliferation and natural killer (NK) cytolytic activity. The acute co-administration of the two drugs resulted in an enhancement of antinociception while they did not yield any additive inhibition of the immune parameters. The CB1 cannabinoid receptor antagonist N-(Piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl 1H-pyrazole-3-carboxamide (SR141716A; 3 mg/kg, i.p.) and the CB2 receptor antagonist N-[(1S)-endo-1,3,3-trimethhyl bicyclo[2.2.1]heptan-2-yl]-5-(4-chloro-3 methylphenyl)-1-(4-methylbenzyl)-pyrazole-3-carboxamide (SR144528; 3 mg/kg, i.p.) did not block the central nor the immune effects of morphine; similarly, the opioid receptor antagonist naloxone did not attenuate CP-55,940-induced effects. Animals tolerant to CP-55,940-induced (0.2 mg/kg, i.p.; twice a day for 4 days) or morphine-induced analgesia (5 mg/kg, s.c.; twice a day for 6 days) also developed tolerance to their acute immunosuppressive effects. Concomitantly, animals became cross-resistant to the immunosuppressive effects while an asymmetric cross-tolerance developed for analgesia. Our data demonstrated the existence of an interaction between cannabinoids and opiates at the immune level that differs from the interaction present in the CNS. PMID- 11431012 TI - Differential effects of neuropeptide Y (NPY) on leukocyte subsets in the blood: mobilization of B-1-like B-lymphocytes and activated monocytes. AB - Sympathetic nervous system activation mobilizes leukocytes but it is unknown whether the concomitant neuropeptide Y (NPY)-release also alters blood leukocyte counts. Using chronic intravenous (i.v.) cannulation of freely moving rats and flow cytometry, time-, dose- and subset-specific effects of NPY on blood leukocytes were investigated 1-15 min after injection: High-dose NPY increases leukocytes numbers by preferentially mobilizing CD4(+) T-cells, activated NKR P1A(+) monocytes and NK-cells. Low-dose NPY significantly decreases B-lymphocyte and NK-cell numbers. Furthermore, NPY dose-dependently mobilizes a previously undetected IgM(low)CD5(+)CD11b(+) B-cell subpopulation in rats ("B1-like" B lymphocytes). These data suggest a role for the sympathetic neurotransmitter NPY in neuroimmune alterations in vivo. PMID- 11431013 TI - Identification of dopamine plasma membrane and vesicular transporters in human peripheral blood lymphocytes. AB - Plasma membrane dopamine transporter (DAT), vesicular monoamine transporters (VMAT) type-1 and -2 and the expression of the dopaminergic markers dopamine and tyrosine hydroxylase were assessed in membranes and/or in cytospin centrifuged human peripheral blood lymphocytes. The radiolabeled DAT ligand [3H]GBR12935 was bound to peripheral lymphocytes in a manner consistent with the specific binding to a dopamine uptake system, with a dissociation constant similar to that found in striatum, but with a lower density of binding sites. On the other hand, no specific binding occurred in cerebellum used as a test tissue not expressing DAT. Western blot analysis using antibodies raised against amino or carboxy terminus of DAT or against VMAT-1 or VMAT-2 revealed labeling of single bands of approximately 76, 55 or 68 KDa, respectively, displaying similar migration characteristics in lymphocytes and test tissues used for comparison. Immunofluorescence revealed that anti-dopamine, anti-tyrosine hydroxylase, anti DAT, anti-VMAT-1 and anti-VMAT-2 antibodies labeled the total population of cytospin-centrifuged lymphocytes mounted on microscope slides. Confocal laser microscopy demonstrated that dopamine and VMAT-2 immunoreactivity was developed mainly in cytoplasmic punctiform areas likely corresponding to vesicles and to a lower extent was associated to plasma membrane. Tyrosine hydroxylase immunoreactivity was diffused to cytoplasm and to plasma membrane of lymphocytes, whereas DAT and VMAT-1 immunoreactivity were located almost exclusively in lymphocyte plasma membrane and cytoplasm, respectively. Lymphocyte DAT characterized in this study has probably functional relevance as [3H]dopamine was taken up by intact lymphocytes and uptake was inhibited specifically by compounds known to affect dopamine transport. These findings indicate that human peripheral blood lymphocytes possess DAT plasma membrane and VMAT-1 and VMAT-2 transporters. Increasing evidence indicates that dopamine transporter changes may be related to neuronal injury. In view of this assessment of lymphocyte DAT and VMAT transporters can be considered for identifying pathologies characterized by impaired dopaminergic neurotransmission. PMID- 11431014 TI - Resistance of CD4-positive T lymphocytes to etoposide-induced apoptosis mediated by upregulation of Bcl-xL expression in patients with HTLV-I-associated myelopathy. AB - Human T-lymphotropic virus type I (HTLV-I)-associated myelopathy (HAM) is characterized by chronic inflammation of the spinal cord. The exact mechanisms that enhance the development of chronic myelopathy remain to be determined. One such mechanism could be an altered response of peripheral blood CD4(+) T lymphocytes to apoptotic stimuli. We examined the sensitivity of these cells to apoptosis in HAM patients and control. Apoptosis was induced by etoposide, which induces mitochondria-dependent apoptosis through the release of cytochrome c from the mitochondria. The percentage of apoptotic cells that expressed hypodiploid DNA among etoposide-treated CD4(+) T lymphocytes was significantly lower in HAM patients than in the control. Western blot analysis of cell lysates derived from CD4(+) T lymphocytes demonstrated that the expression level of Bcl-xL protein was significantly higher in HAM patients than in the control. Our results indicate that peripheral blood CD4(+) T lymphocytes of HAM patients are resistant to apoptosis triggered through mitochondrial death pathway through upregulation of expression of anti-apoptotic protein, Bcl-xL. This phenomenon might contribute to the prolongation and perpetuation of the chronic inflammatory process in the spinal cord of HAM patients. PMID- 11431015 TI - IgG reactivity against citrullinated myelin basic protein in multiple sclerosis. AB - An increased level of citrullinated myelin basic protein (MBP-C8) has been reported in the brains of multiple sclerosis (MS) patients. However, the involvement of the immune response to post-translational modified MBP in the pathophysiology of MS remains speculative. The aim of this study was to compare the levels of immunoglobulin G antibodies to several MBP epitopes, before and after citrullination, in the cerebrospinal fluid (CSF) and sera of MS patients using enzyme-linked immunosorbent assay (ELISA). We analyzed antibody reactivity against various MBP-peptides in the CSF and sera of 60 MS patients, and 30 patients with other neurological diseases (OND) as controls. The peptides tested were: MBP(75-98) (peptide 1), native (peptide 2) and citrullinated (peptide 3) MBP(108-126) (ARG(122)-->Cit(122)), and native (peptide 4) and citrullinated (peptide 5) MBP(151-170) (ARG(159, 170)-->Cit(159, 170)). All selected peptides could support an immune reactivity in CSF and sera of MS and OND patients. A higher reactivity against peptide 4 was found in the CSF of MS patients compared with OND patients (P<0.0001), but not against citrullinated peptides (peptides 3 and 5). However, we observed that the citrullination state of peptide 2 modified the patterns of immune reactivity more markedly in MS patients (P<0.0001) than in OND patients (P<0.02). Although some MBP epitopes could be a potential target in MS, our data did not demonstrate any difference of antibody response to MBP peptides in their citrullinated forms. PMID- 11431016 TI - Oral salbutamol decreases IL-12 in patients with secondary progressive multiple sclerosis. AB - IL-12 is a key cytokine for Th1 cell development and may be important in the pathogenesis of multiple sclerosis (MS). The beta2-agonist salbutamol is known to decrease IL-12 production in monocytes of normal individuals through increased intracellular cAMP. In a prospective open-label study, we investigated by flow cytometry the effect of a 2-week long oral salbutamol treatment on monocyte IL-12 production in 21 secondary progressive MS patients. Baseline IL-12 production was higher in patients than in healthy controls. The treatment induced a significant decrease in the percentage of IL-12-producing monocytes and dendritic cells that lasted up to 1 week after treatment interruption. This first report on the use of salbutamol in MS shows that this drug has immunomodulatory properties both in vivo and in vitro, and may be beneficial in the treatment of MS. PMID- 11431017 TI - Induction of selected chemokines in glial cells infected with Theiler's virus. AB - To elucidate the early events in Theiler's virus-induced demyelination, a model for human multiple sclerosis (MS), chemokine gene activation in the central nervous system (CNS) resident cells upon viral infection was investigated. Viral infection selectively upregulated RANTES and IP-10 gene expression in primary astrocyte cultures and broader chemokine genes in oligodendrocyte and microglia cultures. Both RANTES and IP-10 were stimulated by proinflammatory cytokine interferon-gamma (IFNgamma), but only RANTES by tumor necrosis factor alpha (TNFalpha), suggesting that virus infection induces chemokines overlapping with those inducible by proinflammatory cytokines. These results suggest that glial cells, astrocytes in particular, may be critical for early recruitment of inflammatory cells in the initiation of virus-induced, immune-mediated demyelination. PMID- 11431019 TI - Novel allosteric antagonists shed light on mglu(5) receptors and CNS disorders. AB - Although multiple metabotropic glutamate (mglu) receptor subtypes were cloned in the early 1990s, progress in the characterization of these receptors has been slow because of difficulties in obtaining subtype-selective ligands. However, in the past few years exciting progress has been made on the mglu(5) receptor subtype following the identification of selective non-amino-acid-like ligands that implicate the mglu(5) receptor as a potentially important therapeutic target, particularly for the treatment of pain and anxiety. PMID- 11431020 TI - A new model serves AMPA receptor trafficking. PMID- 11431021 TI - A novel mechanism for the reuptake of CGRP. PMID- 11431022 TI - A role for leptin in autoimmunity? PMID- 11431023 TI - Anti-cancer drug success emerges from molecular biology origins. PMID- 11431024 TI - New targets for Crohn's disease. PMID- 11431025 TI - Placebo effect? PMID- 11431027 TI - Glutamate signalling in the lung. PMID- 11431028 TI - Anandamide: some like it hot. AB - Anandamide was the first endogenous ligand of cannabinoid receptors to be discovered in 1992. Yet, this compound also efficiently activates receptors specific for capsaicin, known as vanilloid type 1 receptors (VR1). Whether anandamide is a physiological VR1 ligand is controversial. However, very recent reports demonstrate that activation of VR1 by anandamide can be significantly enhanced by various regulatory factors, suggesting that this compound might act as an "endovanilloid" under certain conditions. PMID- 11431029 TI - Towards a pharmacology of DNA methylation. AB - DNA methylation plays an important role in controlling gene-expression programs. Increasing evidence indicates that the enzyme responsible for replicating the DNA methylation pattern, DNA methyltransferase 1 (DNMT1), has a role in cancer. In this article, it is suggested that DNMT1 is a multifunctional protein that has regulatory activities in addition to DNA methylation activity. These functions are assembled into one protein to ensure the coordinate replication of DNA and its methylation pattern. The regulatory activities of DNMT1 are proposed to be involved in cellular transformation and should, therefore, serve as the targets for novel anti-cancer agents. PMID- 11431030 TI - Regulation of mglu(7) receptors by proteins that interact with the intracellular C-terminus. AB - The metabotropic glutamate type 7 (mglu(7)) receptor is a widely distributed, mainly presynaptic Group III mglu receptor that can regulate glutamate release. Recently, largely as a result of the identification of specific proteins that interact with the C-terminal domain of this receptor, considerable progress has been made towards understanding some of the mechanisms that underlie the regulation, signal transduction pathways and targeting of mglu(7) receptors. This has led to the proposal that there are three distinct functionally relevant domains present in the intracellular C-terminus of this receptor: (1) a proximal intracellular signalling domain that interacts with G-protein betagamma-subunits and the Ca(2+) sensor Ca(2+)-calmodulin, and is phosphorylated by protein kinase; (2) a central domain thought to provide a signal for axonal targeting; and (3) an extreme PDZ-binding motif that interacts with the protein kinase C interacting protein, PICK1. PMID- 11431031 TI - Therapeutic implications of human endothelial nitric oxide synthase gene polymorphism. AB - Vascular endothelial dysfunction is now recognized as a common phenomenon in an array of cardiovascular disorders. Production of nitric oxide via the endothelial isoform of nitric oxide synthase [eNOS (previously termed NOS3 or ecNOS)] is vital for a healthy endothelium; several polymorphic variations of the gene encoding eNOS (NOS3) are now known and have been investigated with respect to disease risk. Surprisingly, only approximately half of these studies have demonstrated significant associations between NOS3 polymorphisms and cardiovascular disease, and many reports are contradictory. Central issues include adequate statistical power, appropriateness of control cohorts, multigene interactions and plausible biological consequences. So far, the inconsistencies are not unique to the NOS3 polymorphisms, but probably represent the broad challenges in defining genetic aspects of complex disease processes. PMID- 11431032 TI - G-protein-coupled receptors and signaling networks: emerging paradigms. AB - G-protein-coupled receptors (GPCRs) constitute the largest family of cell-surface molecules involved in signal transmission. These receptors play key physiological roles and their dysfunction results in several diseases. Recently, it has been shown that many of the cellular responses mediated by GPCRs do not involve the sole stimulation of conventional second-messenger-generating systems, but instead result from the functional integration of an intricate network of intracellular signaling pathways. Effectors for GPCRs that are independent of G proteins have now also been identified, thus changing the conventional view of the GPCR heterotrimeric-G-protein-associated effector. The emerging information is expected to help elucidate the most basic mechanism by which these receptors exert their numerous physiological roles, in addition to determining why the perturbation of their function results in many pathological conditions. PMID- 11431033 TI - Proteomics--post-genomic cartography to understand gene function. AB - The completion of the genomic sequences of numerous organisms from human and mouse to Caenorhabditis elegans and many microorganisms, and the definition of their genes provides a database to interpret cellular protein-expression patterns and relate them to protein function. Proteomics technologies that are dependent on mass spectrometry and involve two-dimensional gel electrophoresis are providing the main window into the world of differential protein-expression analysis. In this article, the limitations and expectations of this research field are examined and the future of the analytical needs of proteomics is explored. PMID- 11431034 TI - The importance of specific virus diagnosis and monitoring for antiviral treatment. AB - Antiviral treatment, rapid viral diagnosis and point-of-care diagnostics are all relatively new, and their appropriate use not fully appreciated or evaluated. In this article, the need for laboratory diagnosis in relation to antiviral treatment, and practical approaches are discussed, with influenza and herpes simplex virus (HSV) as examples. PMID- 11431035 TI - Regulation of herpes simplex virus type 1 replication in Vero cells by Psychotria serpens: relationship to gene expression, DNA replication, and protein synthesis. AB - Inhibitory effects of ethanolic extracts from seven Chinese herbs on herpes simplex virus type 1 (HSV-1) replication were investigated. From a bioassay guided fractionation procedure, PS-A-6 was isolated from Psychotria serpens (P. serpens), which suppressed HSV-1 multiplication in Vero cells without apparent cytotoxicity. Time-of-addition experiments suggested that the inhibitory action of PS-A-6 on HSV-1 replication was not through blocking of virus adsorption. In an attempt to further localize the point in the HSV-1 replication cycle where arrest occurred, a set of key regulatory events leading to viral multiplication was examined, including viral gene expression, DNA replication, and structural protein synthesis. The results indicated that gB mRNA and protein expression in Vero cells were impeded by PS-A-6. Southern blot analysis showed that HSV-1 DNA replication in Vero cells was arrested by PS-A-6. In addition, PS-A-6 decreased thymidine kinase (tk) and ICP27 mRNA expression in the cells. The mechanisms of antiviral action of PS-A-6 seem to be mediated, at least in part, through inhibition of early transcripts of HSV-1, such as tk and ICP27 mRNAs, arresting HSV-1 DNA synthesis and gB gene expression in Vero cells. Plans are underway for the isolation of pure compounds from PS-A-6 and elucidation of their mechanism of action. PMID- 11431036 TI - Identification and characterization of a benzothiophene inhibitor of herpes simplex virus type 1 replication which acts at the immediate early stage of infection. AB - Analysis of a large compound library in a high throughput virus infection assay screen identified the benzothiophene PD146626 as a potent and specific inhibitor of herpes simplex virus type 1 (HSV-1) replication. PD146626 possessed an EC(50) and EC(90) against HSV-1 of 0.1 and 1 microM, respectively, and mediated no detectable cytotoxicity in cells at concentrations up to 1 microM. Western blot analyses and time of addition experiments demonstrated that in the presence of PD146626 HSV-1 underwent a specific block in viral gene expression at the immediate early stage. However, several observations indicated that a cellular function rather than a viral immediate early transactivator protein represented the molecular target for PD146626, including the lack of resistance of VP16 and ICP0 mutant viruses to the compound, the inability to select resistant strains of HSV-1 following exhaustive serial passaging of virus in the presence of the compound, and the sensitivity of human cytomegalovirus, which lacks VP16 and ICP0 homologs, to the compound. Moreover, kinetic studies suggested an unusual pattern of responsiveness of the host cell to PD146626, in that the compound could induce an extended antiviral state in cells after only a brief exposure. Together these results suggest that PD146626 targets a novel cellular function that is critical for the expression of HSV-1 immediate early genes but not host cell genes. PMID- 11431037 TI - Novel mono- and di-DNA-enzymes targeted to cleave TAT or TAT-REV RNA inhibit HIV 1 gene expression. AB - The regulatory proteins TAT and REV play a very important role in the transcription and replication of HIV-1. In order to seHIV-01lectively down regulate the expression of these genes we synthesized several mono- and one di DNA-enzyme against the TAT or TAT-REV RNA. Several mono-DNA-enzymes possessing the 10-23 catalytic motif were assembled that were targeted to the predicted loop region of TAT or TAT/REV RNA. The cleavage efficiency of each mono-DNA-enzyme was variable and independent of the size of the predicted loop structure of the target RNA. DNA-enzyme targeted against the largest loop region cleaved the substrate RNA poorly. Mono-DNA-enzyme-5944 that targets only the TAT region cleaved the substrate poorly but the DNA-enzyme-5970 that overlaps TAT and REV showed potent cleavage activity. The two DNA-enzymes, when placed in tandem, cleaved the target RNA at multiple sites that were specific for the two mono-DNA enzymes. Only Dz-5970 retained the ability to cleave the target RNA specifically at simulated physiological conditions. They were able to inhibit HIV-1 specific genes efficiently when introduced into a mammalian cell. The extent of inhibition correlated with their cleavage efficiency obtained at standard conditions of cleavage. Although DNA-enzyme-5970 showed the highest reduction (approximately 90%), other DNA-enzymes (mono-DNA-enzyme-5944 and the di-DNA-enzyme) also showed reduction to an extent of 60 and 80% respectively. The inhibitory effect of the DNA-enzyme could be overcome by providing HIV-1 TAT to the cells. PMID- 11431038 TI - Lactoferrin and cyclic lactoferricin inhibit the entry of human cytomegalovirus into human fibroblasts. AB - Lactoferrin is mainly produced by polymorphonuclear leukocytes and has been demonstrated in mammalian milk and external secretions. Lactoferrin is an iron binding, multifunctional protein and may play an important role in immune regulation and in defense mechanisms against bacteria, fungi and viruses. Lactoferricin is a potent antimicrobial peptide generated from the N-terminal part of lactoferrin by pepsin cleavage. We demonstrate that lactoferrins from different species and its N-terminal peptide lactoferricin (particularly the cyclic form) inhibit expression of early and late antigens, as well as production of infectious viral progeny during human cytomegalovirus (HCMV) infection in vitro. Iron-saturated lactoferrin did not affect HCMV antigen expression. Heparin had the same effects as iron-depleted lactoferrin. Yet, mixtures of lactoferrin and heparin did not inhibit HCMV multiplication i.e. lactoferrin and heparin seemed to mutually block each other's antiviral activities. HCMV-infected cells exposed to lactoferrin and cyclic lactoferricin contained less intracellular virus than unexposed cells. The antiviral activity of cyclic lactoferricin was more than seven-fold weaker than that of the maternal molecule. Lactoferrin and cyclic lactoferricin prevented HCMV entrance into the host cell. PMID- 11431040 TI - Blood pressure and dementia: a review. PMID- 11431041 TI - The home hospitalization of frail elderly patients with advanced dementia. PMID- 11431042 TI - Cognitive assessment of severe dementia: the test for severe impairment (TSI). PMID- 11431043 TI - Cognitive and mood disorders in elderly patients with Parkinson's disease. PMID- 11431044 TI - Late onset depression and suicide outcome. PMID- 11431045 TI - Impact on pressure ulcer healing of an arginine-enriched nutritional solution in patients with severe cognitive impairment. PMID- 11431046 TI - Depression in neurological diseases: a review. PMID- 11431047 TI - Early detection and diagnosis of dementia. PMID- 11431049 TI - A study on the validity of different short versions of the geriatric depression scale. PMID- 11431048 TI - The brain of the elderly between normality and pathology: techniques and technologies for the evaluation. PMID- 11431050 TI - Studies on the brain event related potentials in the elderly with white-coat or isolated systolic hypertension. PMID- 11431051 TI - Changes of psychological state in elderly hypertensives with antihypertrensive therapy. PMID- 11431052 TI - Wanderers: features, findings, issues. PMID- 11431053 TI - Dementia special care unit and management of severely demented patients: an experience. PMID- 11431054 TI - Quality assurance: the experience of a geriatric psychiatry day hospital. PMID- 11431055 TI - Sertraline and reboxetine association in depressive pseudodementia: good cardiovascular effects. PMID- 11431056 TI - The search of true links between educational level and dementia. PMID- 11431057 TI - Conselice study: a population based survey of brain aging in a municipality of the Emilia Romagna region: (A.U.S. L. Ravenna). Neuropsychological tests and nutritional status. PMID- 11431058 TI - Deficient folate nutritional status and cognitive performances: results from a retrospective study in male elderly inpatients in a geriatric department. PMID- 11431059 TI - Cognitive deficits in Alzheimer's disease: treatment with acetylcholinesterase inhibitor agents. PMID- 11431060 TI - Sertraline versus small doses of haloperidol in the treatment of agitated behavior in patients with dementia. PMID- 11431061 TI - The relationship between depression and cognition. PMID- 11431062 TI - Risperidone in the treatment of behavioral disorders in elderly patients with dementia. PMID- 11431063 TI - Prevalence of depression in medically hospitalized elderly patients. PMID- 11431064 TI - The importance of the family training in the home care of demented patients. PMID- 11431065 TI - Cognitive deficits in the elderly: a research on environmental effects on psychometrics. PMID- 11431066 TI - The selection of meaningful activities as a treatment for day-care in dementia. PMID- 11431067 TI - Sertonin (5-HT)-related symptoms and fluoxetine in geriatric depression. PMID- 11431068 TI - Genetic background of inflammatory molecules affects protein expression, the risk of developing Alzheimer's disease and cognitive decline. PMID- 11431069 TI - Metamemory and self-perceived health in mild cognitive impairment. PMID- 11431070 TI - Behavioral disturbances in the Alzheimer's care units: a six-months observation. PMID- 11431071 TI - Risk factors for dementia of Alzheimer's type: a case-control, retrospective evaluation. PMID- 11431072 TI - Disability associated with depressive symptoms in elderly primary care attenders. PMID- 11431073 TI - The importance of awareness: an experience of small support groups for the caregivers of Alzheimer's disease patients. PMID- 11431074 TI - Fronto-temporal dementia versus Alzheimer disease. PMID- 11431075 TI - Psychosis, serotonin receptor polymorphism and Alzheimer's disease. PMID- 11431076 TI - Cognitive training in aging and disease (COTRAD): does it work in Italy? PMID- 11431077 TI - Cognitive impairment, insight, depression and suicidal ideation. PMID- 11431078 TI - Impact of decreased frequency of apolipoprotein Eepsilon4 allele on Alzheimer's disease in Southern Italy. PMID- 11431079 TI - Atherogenic risk factors and cognitive function decline in geriatric age. PMID- 11431080 TI - Conselice study: a population based survey of brain aging in a muncipality of the Emilia Romagna region: (A.U.S.L. Ravenna). Design and methods. PMID- 11431081 TI - Oxidative stress and dementia in oldest-old subjects. PMID- 11431082 TI - Living with dementia: is distress influenced by carer personality. PMID- 11431083 TI - Serum zinc levels in demented institutionalized patients. PMID- 11431084 TI - A prototype system for resource allocation related to the actual disability and comorbidity of patients. PMID- 11431085 TI - Prevalence of depression and cognitive deficit in a geriatric population. PMID- 11431086 TI - Analysis of individual items of mini-mental state examination in discrimination between normal and demented subjects. PMID- 11431087 TI - Twenty-four-hour urinary free cortisol levels in vascular dementia and in Alzheimer's disease. PMID- 11431088 TI - Lipoprotein(a), vascular dementia and Alzheimer's disease. PMID- 11431089 TI - An investigation on behavioral problems in centenarians. PMID- 11431090 TI - 'Janus face' of nitric oxide action on plasma membrane and intracellular ionic channels. PMID- 11431091 TI - A case of occipital epilepsy in and elderly woman. PMID- 11431092 TI - Therapeutic aspects of depression in the elderly. PMID- 11431093 TI - Cognitive style, cognitive impairment and health status in elderly. PMID- 11431094 TI - Determinants of functional status in Alzheimer's disease and vascular dementia. PMID- 11431095 TI - Cognitive disorders in the elderly. Genetic and epigenetic aspects. PMID- 11431096 TI - Tumor necrosis factor-alpha and mood disorders in the elderly. PMID- 11431097 TI - Factors associated with dementia in subjects with cerebrovascular disease. PMID- 11431098 TI - Trk receptor tyrosine kinases: a bridge between cancer and neural development. AB - The proto-oncogene Trks encode the high-affinity receptor tyrosine kinases for neurotrophins of a nerve growth factor (NGF) family. The Trk signals spatiotemporally regulate neural development and maintenance of neural network. However, Trk was originally cloned as an oncogene fused with the tropomyosin gene in the extracellular domain. Accumulating evidence has demonstrated that the rearranged Trk oncogene is often observed in non-neuronal neoplasms such as colon and papillary thyroid cancers, while the signals through the receptors encoded by the proto-oncogene Trks regulate growth, differentiation and apoptosis of the tumors with neuronal origin such as neuroblastoma and medulloblastoma. The intracellular Trk signaling pathway is also different depending on the Trk family receptors, cell types and the grade of transformation. Furthermore, developmentally programmed cell death of neuron, which is largely regulated by neurotrophin signaling, is at least in part controlled by tumor suppressors p53 and p73 as well as their antagonist DeltaNp73. Thus, the Trks and their downstream signaling function in both ontogenesis and oncogenesis. In this short review, the dynamic role of the Trk family receptors signaling in neural development, neurogenic tumors and other cancers will be discussed. PMID- 11431099 TI - Dual role of vitamin C on lipid profile and combined application of cyclophosphamide, methotrexate and 5-fluorouracil treatment in fibrosarcoma bearing rats. AB - Combined application of cyclophosphamide, methotrexate, 5-fluorouracil (CMF) has been followed in the treatment of breast cancer. The combined effect of CMF and vitamin C on plasma lipid and lipoprotein is important, since vitamin C encumbers the lipid abnormalities instigated by CMF. Hence, the study was launched to appraise the salubrious role of vitamin C in CMF administered fibrosarcoma bearing rats. Fibrosarcoma cell line-induced rats were treated with CMF (cyclophosphamide 10 mg/kg b.w., methotrexate 1 mg/kg b.w., 5-fluorouracil 10 mg/kg b.w. and vitamin C 200 mg/kg b.w.) individually and in combination for 120 days. The concentration of plasma lipids and lipoprotein was determined in control and experimental rats. The untreated as well as CMF administered fibrosarcoma-bearing rats divulged significantly in increased levels of plasma total cholesterol, triglycerides, phospholipids, very low density lipoprotein (VLDL) and low density lipoprotein (LDL) cholesterol, as compared with their respective control animals. Whereas ester and high density lipoprotein (HDL) cholesterol levels exhibited a marked decrease in these animals. However, these lipid abnormalities were found to be moderated by co-administration of vitamin C. These results suggested that some clinical entanglement of CMF was refrained by co-administration of vitamin C in tumor stress condition. PMID- 11431100 TI - Effect of bile acids on formation of azoxymethane-induced aberrant crypt foci in colostomized F344 rat colon. AB - The effect of bile acids on the formation of azoxymethane induced aberrant crypt foci (ACF) was investigated using the fecal stream-excluded colons of colostomized F344 rats. The excluded colon was irrigated with saline or bile acids (1 mg/0.5 ml per day, 5 days/week) for 4 weeks. The mean numbers of ACF per colon in rats given cholic acid, deoxycholic acid (DCA), chenodeoxycholic acid (CDCA), lithocholic acid, and ursodeoxycholic acid (UDCA) were 160.8, 118.2, 227.8, 150.7 and 87.3, respectively, while that of the control was 174.0. The number of ACF was significantly larger in CDCA, but smaller in UDCA and DCA treated rats than the control (P<0.01). DCA did not induce apoptosis in the colon under the present conditions. PMID- 11431101 TI - Cancer initiation by fumonisin B(1) in rat liver--role of cell proliferation. AB - Fumonisin B(1) (FB(1)), a carcinogenic mycotoxin produced by the fungus Fusarium verticillioides in corn, causes cancer initiation in rat liver in a similar manner to genotoxic carcinogens although apparently with different kinetics. The present experiment was designed to evaluate the role of regenerative cell proliferation, effected by partial hepatectomy (PH) and carbontetrachloride (CCl(4)) and direct mitogen-induced hyperplasia, induced by lead nitrate (PbNO(3)), on FB(1)-induced cancer initiation. Initiation was effected over a period of 14 days by gavage administration of FB(1) at different daily doses ranging from 0.14 to 3.5 mg FB(1)/100 g body weight while the stimuli for cell proliferation were introduced 7 days after the start of the FB(1) treatment. Based on the proliferative stimulus used, cancer promotion was effected 3 weeks after completion of the initiating treatment by 2-acetylaminofluorene (2-AAF) treatment followed by PH or carbon tetrachloride CCl(4) on day 4. Cancer initiation by FB(1) was associated with a hepatotoxic effect and an increase in lipid peroxidation. In contrast to compensatory liver cell proliferation induced by PH and CCl(4), mitogen-induced hyperplasia (PbNO(3)) failed to enhance the cancer initiating potential of FB(1) suggesting that cancer induction by a non genotoxic carcinogen is supported by regenerative cell proliferation. Cognizance of the enhancing role of cell proliferation during cancer initiation by FB(1) is required in assessing the risks posed by this mycotoxin to humans. PMID- 11431102 TI - Oxidative DNA damage by capsaicin and dihydrocapsaicin in the presence of Cu(II). AB - Capsaicin is the pungent phenolic principle of the Capsicum species, and has shown a wide range of pharmacological properties, including antigenotoxic, antimutagenic, and anticarcinogenic effects. Other studies have, however, shown it to be a tumor promoter and potential mutagen, and a carcinogen, resulting in capsaicin being termed a 'double edged sword'. In the present study, we show that capsaicin is capable of causing strand scission in calf thymus and plasmid DNA in the presence of Cu(II) and that this breakage is mediated by reactive oxygen species, especially the hydroxyl radical. Our results further show that capsaicin can directly generate hydroxyl radicals in the presence of Cu(II). To explore the chemical basis of the DNA breakage reaction by capsaicin, we have compared these properties of capsaicin with its saturated structural analog dihydrocapsaicin (DHC). The rate of DNA degradation, as well as hydroxyl radical formation, was found to be greater in the case of capsaicin. Both capsaicin and DHC are able to reduce Cu(II) to Cu(I), which was shown to be an essential intermediate in the DNA cleavage reaction. Stoichiometric analysis indicated that whereas 1 mol of capsaicin reduced 3 mol of Cu(II), 1 mol of DHC reduced only 2 mol of Cu(II). This explains the greater activity of capsaicin and also leads to a model for copper binding to the capsaicins. PMID- 11431103 TI - Post-initiation treatment of Indole-3-carbinol did not suppress N-methyl-N nitrosourea induced mammary carcinogenesis in rats. AB - The consumption of cruciferous vegetables (the Family of Cruciferae) such as cabbage, broccoli and Brussels sprouts has been shown to have cancer chemopreventive effects in humans and experimental animals. Indole-3-carbinol (I3C), one component of cruciferous vegetables, has been shown to exert cancer chemopreventive influence in liver, colon, and mammary tissue when given before or concurrent with exposure to a carcinogen. However in some reports, there has been evidence that consumption of I3C after carcinogen treatment might be associated with tumor promotion in some tissues. There have been no reports, to our knowledge, of post-initiation effects of I3C in the N-methyl-N-nitrosourea (MNU)-induced mammary tumor model in rats. Our studies were performed to examine this question. Ninety-six, 4-week-old female Sprague-Dawley rats were randomly divided into five groups. The animals of groups 1, 2 and 3 received an intraperitoneal injection of MNU at the age of 50 days. The animals of groups 4 and 5 were injected with saline only at the same time. Animals of groups 1 and 2 were given diet containing 100 ppm and 300 ppm I3C from week 1 until week 25 after MNU treatment. The animals of group 4 were given basal diet containing 300 ppm I3C without MNU treatment. All animals were killed at week 25. The incidences of mammary tumors in the groups 1, 2 and 3 were 95.8% (23/24), 83.3% (20/24) and 82.4% (28/34), respectively. The average number of tumors in the tumor bearing rats of the MNU and I3C 300 ppm group (group 2; 3.85+/-0.63) was higher than that in the MNU alone group (group 3; 2.46+/-0.31). These results represented that exposure to I3C after carcinogen treatment did not suppress development of mammary tumors. PMID- 11431104 TI - DNA demethylase is expressed in ovarian cancers and the expression correlates with demethylation of CpG sites in the promoter region of c-erbB-2 and survivin genes. AB - The objectives of this study were to examine DNA demethylase (dMTase) expression in ovarian cancers and evaluate methylation of CpG sites in the promoter of the c erbB-2 gene and survivin gene exon 1. Forty-three epithelial ovarian cancers and 43 non-cancerous ovarian tissues were studied for dMTase expression by RT-PCR. Genomic DNA was extracted and digested with HindIII and then HpaII. CpG site sensitive primers were constructed to amplify the promoter of the c-erbB-2 gene and survivin gene exon 1. Immunohistochemical evaluation of ErbB-2 protein and RT PCR for survivin were also performed. dMTase was positive in 88.4% of ovarian cancers but only in 9.3% of non-cancerous ovaries (P<0.001, Fisher's exact test). The expression was similarly observed in both early stage (stage I+II: 17/19) and advanced stage (stage III+IV: 21/24) groups of ovarian malignancy. It was found that 78.9% of dMTase-positive cancers had both c-erbB-2 promoter and survivin gene exon 1 unmethylated, whereas 40% of dMTase-negative cancers had both sites methylated. In non-cancerous ovaries, these sites were mostly methylated (90.6%) and the difference from cancer cases was highly significant (P<0.001). Immunohistochemical evaluation of ErbB-2 showed significant correlation of unmethylated c-erbB-2 promoter and ErbB-2 expression. The RT-PCR for survivin expression showed that 86% of cancers were positive and six cases were negative. Exon 1 was methylated in 83% of the survivin-negative cases. This is the first report of dMTase expression in ovarian cancers. The correlation of dMTase expression with unmethylation of c-erbB-2 promoter and survivin gene exon 1 suggests that these sites may be targets for demethylation by the enzyme. The up regulation of oncogenes may be the consequence of epigenetic control of gene expression by the dMTase. PMID- 11431105 TI - Cancer procoagulant and blood platelet activation. AB - The effects of cancer procoagulant (CP), cysteine protease (EC 3.4.22.26), on the pig blood platelet secretory process and platelet aggregation have been studied. The response of platelets to CP was compared with the response of these cells to thrombin. The obtained results show that blood platelets treated with CP (0.5, 1, 2.5, and 5 microg/ml, 2-30 min, 37 degrees C) released adenine nucleotides (P < 0.05) and proteins (P < 0.05). The secretion of compounds from blood platelets after incubation with CP does not correlate with the release of platelet lactic dehydrogenase activity (marker of cell lysis) into the extracellular medium. In comparison with thrombin action, CP stimulates secretory process to a smaller extent than thrombin alone. In the presence of CP, the thrombin action is suppressed (P < 0.05). We noticed that CP does not induce platelet aggregation. PMID- 11431106 TI - Expression of protease-activated receptor 1 in oral squamous cell carcinoma. AB - Protease-activated receptor 1 (PAR-1) is a G-coupled membrane protein. In this study, we analyzed the expression of PAR-1 in oral squamous cell carcinomas (SCCs). PAR-1 was expressed in oral SCCs, but the level of PAR-1 protein was lower in non-metastatic cells than in metastatic cells. Thrombin stimulated the growth of metastatic cells, and both thrombin and thrombin receptor activation peptide (TRP) enhanced the adhesion of these cells to fibronectin, but had no effect on non-metastatic cells. Thrombin and TRP also induced matrix metalloproteinase (MMP)-2 and MMP-9 activities in metastatic cells. These results suggest that PAR-1 may contribute to the growth and invasive potential of oral SCC. PMID- 11431107 TI - To predict chemotherapy response using technetium-99m tetrofosmin and compare with p-glycoprotein and multidrug resistance related protein-1 expression in patients with untreated small cell lung cancer. AB - The aim of this study was to investigate the relationships among technetium-99m tetrofosmin (Tc-TF) accumulation in untreated small cell lung cancer (SCLC), the expression of P-glycoprotein (Pgp) and multidrug resistance related protein-1 (MRP1), and the response to chemotherapy in patients with untreated SCLC. Thirty patients with SCLC were studied with chest scintigraphy 15 to 30 min after intravenous injection of Tc-TF before chemotherapeutic induction. Tc-TF chest scans were interpreted both visually and quantitatively. The response to chemotherapy was evaluated upon completion of chemotherapy. Immunohistochemical analyses were performed on multiple non-consecutive sections of biopsy specimens to detect Pgp and MRP1 expression. Fifteen patients with good response to chemotherapy had a significantly higher incidence (100.0%) of positive Tc-TF chest single photon emission computed tomography (SPECT) findings and negative Pgp or MPR expression than 15 patients with poor response (20%) (P<0.05). The tumor/background (T/B) ratios were 1.8+/-0.3 and 1.2+/-0.3 for patients with good response and poor response, respectively (P<0.05). However, other prognostic factors (performance status, tumor size and stage) were not significantly related to Tc-TF chest scan findings and response to chemotherapy. Tc-TF chest scintigraphy correlated well with Pgp or MRP1 expression and accurately predicted the response to chemotherapy in patients with SCLC. PMID- 11431108 TI - Phenotypic alterations in Caki-1 cells as a consequence of TIMP-1 overexpression. AB - Maintenance of the extracellular matrix (ECM) is important for tissue integrity and cellular physiology. Normal ECM turnover is regulated by a balance between matrix metalloproteinases and their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). In metastasis, this balance favours increased ECM degradation. The objective of this study was to determine the effects of TIMP-1 overexpression on the metastatic process. To this end, we stably transfected a renal carcinoma cell line, Caki-1, with TIMP-1, using a pRc/CMV expression plasmid and LIPOFECTAMINE transfection reagent. The resultant clones displayed increased adhesion on the ECM substratum, including collagen type IV and laminin, and altered invasive capacity through fibronectin and Matrigel, dependent upon the level of TIMP-1 expression. These changes were not due to altered integrin expression, as assessed by flow cytometry. As well as protease inhibitory activity, TIMPs can influence cell proliferation and cell survival. The TIMP-1 clones displayed no changes in proliferation under normal growth conditions, compared with Caki-1 cells. However, under reduced serum conditions, the TIMP-1 clones had a greater percentage of cells in both S (P<0.05) and G(2)/M (P<0.005) phases and less cells in G(0)/G(1) (P<0.001) of the cell cycle than Caki-1 cells. The results confirm a dual role for TIMP-1 in invasion and metastasis, and provide further clues behind the molecular mechanisms in these processes. PMID- 11431120 TI - Multiple-label immunocytochemistry for the evaluation of nature of cell death in experimental models of neurodegeneration. AB - A prominent feature of neurodegenerative diseases is a loss of specific neuronal populations. The pathophysiological mechanisms responsible are, however, poorly understood. Primary cultures of rodent embryonic neurons represent a useful experimental system for investigation of molecular pathways of neurodegeneration and mechanisms of cell death. Here, we report a technique utilizing triple-label immunocytochemistry with confocal immunofluorescence detection designed to simultaneously assess multiple parameters of cell injury in individual hippocampal neurons in primary culture. This method combines detection of DNA damage (TUNEL or Klenow assay) with double-label immunocytochemistry for the activated form of caspase-3 or, alternatively, caspase-cleaved actin (fractin), and microtubule-associated protein-2 (MAP-2) or beta-tubulin. The combined evaluation of the form of nuclear damage (karyorrhexis, pyknosis), the presence or absence of activated caspase-3, and the extent of the damage to cell cytoskeleton, allows for precise assessment of the extent of injury and the mode of cell death (apoptosis, oncosis) for individual neurons. PMID- 11431121 TI - Motor, cytoarchitectural and biochemical assessment of pharmacological neuroprotection against CNS damage induced by neonatal exposure to ionizing radiation. AB - Exposure of neonatal rats to a 5 Gy single dose of X-irradiation induces permanent abnormalities in cerebellar cortex cytoarchitecture and neurochemistry and motor function. This rodent model constitutes an useful tool to evaluate morphological, neurochemical and motor changes induced by ionizing radiation and the possible restorative effects of potential or clearly established neuroprotective drugs. After selection and administration of a neuroprotective agent to neonatally irradiated rats, quantitative evaluations of motor behavior (gait), cerebellar cortex cytoarchitecture and cerebellar monoamine levels are performed. Data are compared to those of both saline-injected, X-irradiated, and saline-injected, sham-irradiated controls. Evaluation of data from the different experimental groups is performed at postnatal days 30 and 90. After this postnatal interval, radiation-induced damage of cerebellar function in nonprotected rodents is considered to be permanent. The longitudinal evaluation of various parameters in the different experimental groups through a multidisciplinary approach, allows determination of the variables that are more sensitive to X-irradiation-induced damage and/or neuroprotective agent-induced restoration. Given the well-known correspondence in cerebellar developmental stages between rodents and humans, this model and related studies bring health related implications, considering the accidental or therapeutic exposure of developing human beings to ionizing radiation. PMID- 11431122 TI - How to use the optical fractionator: an example based on the estimation of neurons in the hippocampal CA1 and CA3 regions of tree shrews. AB - Until recently, exposure of the hippocampus to prolonged elevated glucocorticoid levels was thought to result in damage and loss of pyramidal neurons. Most of the earlier studies were based on measures of neuronal density and used assumptions based counting methods. Using a stereological technique, the optical fractionator, which eliminates potential biases inherent in the assumption-based techniques, we were able to demonstrate that chronic psychosocial stress in tree shrews has no effect on neuronal number in the hippocampal CA1 and CA3 regions. The present report will focus on the practical aspects of the optical fractionator, by describing in detail how to estimate the total number of neurons in the hippocampal CA1 and CA3 regions of tree shrews. In this example the group sizes have been increased over those used in the earlier study. The present study supports our previous conclusion that stress does not affect the number of hippocampal neurons in the CA1 and CA3 areas as suggested by other authors. The results obtained with the optical fractionator can be used to estimate the precision of the data. PMID- 11431123 TI - Elucidation of neuronal circuitry: protocol(s) combining intracellular labeling, neuroanatomical tracing and immunocytochemical methodologies. AB - We describe a protocol combining either intracellular biotinamide staining or anterograde biotinylated dextran amine (BDA) tracing with retrograde horseradish peroxidase (HRP) labeling and immunocytochemistry in order to map physiologically identified neuronal pathways. Presynaptic neurons including their boutons are labeled by either intracellular injection of biotinamide or extracellular injection of BDA while postsynaptic neurons are labeled with HRP via retrograde transport. Tissues are first processed to detect HRP using a tetramethylbenzidine and sodium-tungstate method. Biotinamide or BDA staining is then visualized using an ABC-diaminobenzidine-Ni method and finally the tissue is immunocytochemically stained using choline acetyltransferase (ChAT) or parvalbumin antibodies and a peroxidase-anti-peroxidase method. After processing, biotinamide, BDA, HRP and immunocytochemical staining can readily be distinguished by differences in the size, color and texture of their reaction products. We have utilized this methodology to explore synaptic relationships between trigeminal primary afferent neurons and brainstem projection and motoneurons at both the light and electron microscopic levels. This multiple labeling methodology could be readily adapted to characterize the physiological, morphological and neurochemical properties of other neuronal pathways. PMID- 11431124 TI - Voltage-clamp recordings of postsynaptic currents in substantia gelatinosa neurons in vitro and its applications to assess synaptic transmission. AB - We describe here procedures for recording postsynaptic currents in substantia gelatinosa neurons on a transverse spinal cord slice preparation with an attached dorsal root. At the holding potential of -70 mV, glutamatergic spontaneous excitatory postsynaptic currents (EPSCs) and dorsal root (A delta and/or C fiber) stimulation-evoked EPSCs could be observed. Whereas at the holding potential of 0 mV, spontaneous inhibitory postsynaptic currents (IPSCs) and dorsal root A delta fiber stimulation-evoked IPSCs could be encountered. The methods make it possible to evaluate synaptic transmission by analysing the postsynaptic currents on dorsal root attached spinal cord slice. PMID- 11431125 TI - Four paradigms to study visual--spatial attention of myopic subjects. AB - We describe four protocols for evaluating the attentional skills of myopic and control subjects in target stimulus detection tasks; simple reaction times (RT) are recorded. Two protocols are designed to study both automatic and voluntary orienting of attention. Modified implicit orienting paradigms [Q. J. Exp. Psychol. 32A (1980) 2; J. Exp. Psychol. Gen. 109 (1980) 160] are used in which cues elicit a shift of attention but gaze is maintained on a central fixation point. A third paradigm is designed to study the time-course of focusing; two circles (focusing cues) of different diameters are randomly presented on a point of the visual space where attention has been previously shifted. Seven SOAs (stimulus onset asynchrony) are used. The fourth paradigm was designed to evaluate visual search; three circular arrays of four, eight and twelve stimuli are randomly presented around a fixation point and subjects have to detect the target stimulus inside one of the circles (the other circles are distracters). Since some attentional deficits are associated with myopia [Invest. Ophthalmol. Vis. Sci. 38 (1997) 1725; Cogn. Brain Res. 8 (1999) 369], these protocols could be useful both in the detection of deficits in subjects on the verge of becoming myopics and in the development of cognitive training programs to reduce attentional deficits. PMID- 11431126 TI - Preparation and handling of brain mitochondria useful to study uptake and release of calcium. AB - There is increasing evidence for a critical role of mitochondria in calcium homeostasis and neuronal death in excitotoxicity. In spite of much work during the last two decades, the kinetic parameters of Ca(2+) transport in brain mitochondria remain controversial. Analysis of the literature data suggests that these contradictions can be due to differences in the methodology used to prepare or to incubate brain mitochondria. In the present communication, the whole protocol for preparation of non-synaptic rat forebrain mitochondria is described. This report shows that this preparation is well coupled and essentially free of non-mitochondrial contaminants. The mitochondria obtained are useful to study Ca(2+) uptake and release. Both Na(+)-independent, Na(+)-dependent and spontaneous Ca(2+) release may be studied with this preparation. This system is also useful in studies on the role of mitochondria and other intracellular Ca(2+) stores in disturbance of Ca(2+) homeostasis and delayed cell death under excitotoxic conditions. PMID- 11431127 TI - Transection of the optic nerve in rats: studying neuronal death and survival in vivo. AB - Transection of the optic nerve (ON) in the adult rat, as a model of fiber tract lesion in the adult mammalian CNS, results in delayed, mainly apoptotic death of 80--90% of retinal ganglion cells (RGCs) within 14 days post-lesion. Because of good surgical accessibility of the retina and the optic nerve, the retino-tectal projection represents not only a convenient model to study the molecular mechanisms underlying neuronal death but also serves as a suitable system for investigating potential neuroprotective agents in vivo. In the present report, we provide a detailed protocol for this model including retrograde labeling of RGCs, ON lesion, assessment of the number of surviving neurons, and tissue preparation for several standard techniques like immunohistochemistry, reverse transcription- polymerase chain reaction (RT--PCR), enzyme assays and Immunoblot. PMID- 11431128 TI - A device for stereotactic transection of fiber bundles in rats. AB - We have designed a device for stereotactic transection of fiber bundles in experiments using rats. Here, we present our assessment of its features relative to those of conventional methods. The instrument consists of a stainless steel cannula and a thin inner wire with a hook at one end and a hilt at the other. The hook can be extended or withdrawn freely by pushing or pulling the hilt. Lesions were evaluated in 12 male Wistar rats, after two targets, the anterior commissure (n=6) and the caudate-putamen (n=6), were transected. After the cannula was introduced into the target, the inner hook was extended in an anterior direction orthogonally to the transverse plane. Next, the entire device was pulled back along the insertion path to transect the neuronal fibers. Then the inner hook was withdrawn into the cannula and the entire device was removed. Seven days later, brains were removed for histologic processing. Microscopic examination demonstrated a slit like gap produced by transection at the target; the lesions were infiltrated by microglia and surrounded by gliosis. Adjacent regions were minimally damaged. The path of the cannula demonstrated only minimal gliosis. Unlike conventional methods, this device permits precise transection of deep fiber bundles with minimal damage to surrounding brain tissue. PMID- 11431129 TI - Monitoring NAD(P)H autofluorescence to assess mitochondrial metabolic functions in rat hippocampal-entorhinal cortex slices. AB - Changes in neuronal energy metabolism, mitochondrial functions and homeostasis of reactive oxygen species are often supposed to induce alterations in neuronal activity in hippocampal slice models. In order to investigate the NAD(P)H autofluorescence signal in brain slice models, methods to monitor NAD(P)H signal in isolated mitochondria as described by Chance et al. [J. Biol. Chem. 254 (1979) 4764] and dissociated neurons as described by Duchen [Biochem. J. 283 (1992) 41] were adapted to recording conditions required for brain slices. Considering different experimental questions, we established an approach to monitor NAD(P)H autofluorescence signals from hippocampal slices of 400 microm thickness under either submerged or interface conditions. Therefore the procedure described here allows the measurement of NAD(P)H autofluorescence under conditions typically required in electrophysiological experiments. Depolarization of plasma membrane caused by electrical stimulation or application of glutamate (100 microM) resulted in a characteristic initial decrease followed by a long-lasting increase in the NAD(P)H autofluorescence signal. H(2)O(2) (100 microM) evoked a strong NAD(P)H signal decrease indicating direct oxidation to the nonfluorescencend NAD(P)(+). In contrast, the increase in NAD(P)H signal that followed a brief inhibition of mitochondrial respiratory chain complex I using rotenone (1 microM) indicated an accumulation of NAD(P)H. However, in presence of rotenone (1 microM) electrically evoked long-lasting NAD(P)H signal overshoot decreased progressively, due to a negative feedback of accumulated NAD(P)H to the citrate cycle. A comparable reduction in NAD(P)H signal increase were observed during low Mg(2+) induced epileptiform activity, indicating a relative energy failure. In conclusion, the method presented here allows to monitor NAD(P)H autofluorescence signals to gain insight into the coupling of neuronal activity, energy metabolism and mitochondrial function in brain slice models. PMID- 11431130 TI - Simultaneous determination of multiple transcripts and splice variants of a primary transcript using ribonuclease protection assays. AB - The ribonuclease (RNase) protection assay (RPA) is an extremely sensitive technique used to determine specific mRNAs from cell and tissue extracts. The present protocol presents detailed procedures for a conventional RPA using antisense RNA probes purified with a Fullengther apparatus. The Fullengther has the advantage of being a relatively quick and safe procedure compared to more conventional methods for purification of full-length RNA probes. Using this protocol, we sought to simultaneously determine multiple mRNA species, including splice variants of the type I receptor (PAC(1)) of pituitary adenylate cyclase activating polypeptide (PACAP), an important mediator in the regulation of luteinizing hormone-releasing hormone (LHRH) synthesis by ovarian steroids such as progesterone [7]. PAC(1) has more than eight splice variants. We have been able to discriminate the hop1 variant from other splice variants. To improve our understanding of the regulation mechanism of genes that are related to each other, such as LHRH and PACAP, it is most important to simultaneously determine genes that are involved in the same physiological areas of regulation. Using only 5 microg of total RNA sample from a single rat preoptic area, we simultaneously determined five different transcripts, including four rare mRNA species such as LHRH, PACAP, and hop1 variant and other splice variants of PAC(1), as well as the internal control of cyclophilin mRNA. This protocol provides a method for the simultaneous determination of multiple transcripts using the RPA. PMID- 11431132 TI - The effects of intense exercise on the female reproductive system. AB - Women have become increasingly physically active in recent decades. While exercise provides substantial health benefits, intensive exercise is also associated with a unique set of risks for the female athlete. Hypothalamic dysfunction associated with strenuous exercise, and the resulting disturbance of GnRH pulsatility, can result in delayed menarche and disruption of menstrual cyclicity. Specific mechanisms triggering reproductive dysfunction may vary across athletic disciplines. An energy drain incurred by women whose energy expenditure exceeds dietary energy intake appears to be the primary factor effecting GnRH suppression in athletes engaged in sports emphasizing leanness; nutritional restriction may be an important causal factor in the hypoestrogenism observed in these athletes. A distinct hormonal profile characterized by hyperandrogenism rather than hypoestrogenism is associated with athletes engaged in sports emphasizing strength over leanness. Complications associated with suppression of GnRH include infertility and compromised bone density. Failure to attain peak bone mass and bone loss predispose hypoestrogenic athletes to osteopenia and osteoporosis. Metabolic aberrations associated with nutritional insult may be the primary factors effecting low bone density in hypoestrogenic athletes, thus diagnosis should include careful screening for abnormal eating behavior. Increasing caloric intake to offset high energy demand may be sufficient to reverse menstrual dysfunction and stimulate bone accretion. Treatment with exogenous estrogen may help to curb further bone loss in the hypoestrogenic amenorrheic athlete, but may not be sufficient to stimulate bone growth. Treatment aimed at correcting metabolic abnormalities may in fact prove more effective than that aimed at correcting estrogen deficiencies. PMID- 11431133 TI - Insulin, growth hormone and sport. AB - This review examines some interesting 'new' histories of insulin and reviews our current understanding of its physiological actions and synergy with GH in the regulation of metabolism and body composition. It reviews the history of GH abuse that antedates by many years the awareness of endocrinologists to its potent anabolic actions. Promising methods for detection of GH abuse have been developed but have yet to be sufficiently well validated to be ready for introduction into competitive sport. So far, there are two promising avenues for detecting GH abuse. The first uses immunoassays that can distinguish the isomers of pituitary derived GH from the monomer of recombinant human GH. The second works through demonstrating circulating concentrations of one or more GH-sensitive substances that exceed the extremes of normal physiological variability. Both methods require blood rather than urine samples. The first method has a window of opportunity lasting about 24 h after an injection and is most suitable for 'out of competition' testing. The second method has reasonable sensitivity for as long as 2 weeks after the last injection of GH and is uninfluenced by extreme exercise and suitable for post-competition samples. This method has a greater sensitivity in men than in women. The specificity of both methods seems acceptably high but lawyers need to decide what level of scientific probability is needed to obtain a conviction. Both methods need further validation before implementation. Research work carried out as part of the fight against doping in sport has opened up a new and exciting area of endocrinology. PMID- 11431134 TI - Proof of the effect of testosterone on skeletal muscle. AB - In spite of the widespread abuse of androgenic steroids by athletes and recreational body-builders, the effects of these agents on athletic performance and physical function remain poorly understood. Experimentally induced androgen deficiency is associated with a loss of fat-free mass; conversely, physiologic testosterone replacement of healthy, androgen-deficient men increases fat-free mass and muscle protein synthesis. Testosterone supplementation of HIV-infected men with low testosterone levels and of older men with normally low testosterone concentrations also increases muscle mass. However, we do not know whether physiologic testosterone replacement can improve physical function and health related quality of life, and reduce the risk of falls and disability in older men or those with chronic illness. Testosterone increases maximal voluntary strength in a dose-dependent manner and thus might improve performance in power-lifting events. However, testosterone has not been shown to improve performance in endurance events. The mechanisms by which testosterone increases muscle mass are not known, but probably involve alterations in the expression of multiple muscle growth regulators. PMID- 11431135 TI - Behavioural effects of androgen in men and women. PMID- 11431136 TI - Ethical aspects and the prevalence of hormone abuse in sport. AB - Abuse of hormones by sportsmen or sportswomen might lead to some serious health consequences for the individual user. Such abuse also damages the very spirit of sport, cheating fellow athletes, the officials and spectators. Where hormone findings cause most controversy is with endogenously produced substances. Some hormone findings might be indicators of a medical condition, and it requires sensitive handling to discover the facts. Examining the prevalence of hormone abuse using a theoretical perspective on ethics permits a philosophical study of the dilemmas facing sport, and a clearer identification of the issues that science can help sport to resolve. This paper looks at the way in which rules on doping have evolved in an attempt to set out the ethical standards that should apply and to discuss how some sportsmen and sportswomen have worked around the rules, challenging them to the extent that the anti-doping system itself is questioned. The data emerging from the testing programmes give one guide to the actual prevalence of hormone findings. However, as not all findings may constitute doping offences this cannot be said to be the definitive guide to the extent of hormone abuse. Use of hormone medications with or without a therapeutical indication further complicates the disciplinary process. Sensitive management of hormone findings is absolutely necessary to avoid accusations of doping or embarrassing breaches of confidentiality when the origin may be a serious medical condition. Because hormone findings require careful consideration, the door is open for the anti-doping system to be exploited by unscrupulous scientists, raising challenges that test the limits of credibility. Sex, alcohol and decomposition are arguments that have been put forward to explain findings. A close partnership between scientists and sport is called for to avoid the athlete becoming a victim of the rules that are intended to protect sport. PMID- 11431137 TI - Drugs in sport - the role of the physician. AB - Sportsmen have used anabolic steroids since the 1950s and yet it was not until the 1980s that we, as physicians, admitted that they could improve performance. We now find ourselves in the insidious position of being unable to predict convincingly either safety or major health risks with performance-enhancing drug use. The use of performance-enhancing drugs is no longer limited to the elite athlete. In 1993 the Canadian Center for Drug-free Sport estimated that 83 000 children between the ages of 11 and 18 had used anabolic steroids in the previous 12 months. Recent evidence suggests anabolic steroids are now the third most commonly offered drugs to children in the UK, behind cannabis and amphetamines. The role of the physician of today is to regain our position of impartiality and objectivity within both the sporting and general community. Only then will we be able to pursue a harm minimisation strategy designed to convince the public that it is better to be the best you can be naturally. For the majority, the improvement through the use of performance-enhancing drugs can equally be achieved through dietary and training advice. For the elite athlete, what price a gold medal that is tarnished by deceit? Its value then can only lie with the sponsors and politicians, for they can no longer claim to be sportsmen, only entertainers. PMID- 11431138 TI - The acid-labile subunit (ALS) of the 150 kDa IGF-binding protein complex: an important but forgotten component of the circulating IGF system. AB - The insulin-like growth factors-I and -II (IGFs) are involved in a wide array of cellular processes such as proliferation, prevention of apoptosis, and differentiation. Most of these effects are mediated by the IGF-I receptor, although at higher IGF concentrations the insulin receptor can also be activated. As the expression of both the IGFs and their receptors is widespread, IGFs are thought to have autocrine/paracrine modes of actions also, particularly during foetal life. The endocrine component of the IGF system is recognised to be important after birth, with IGF-I mediating many of the effects of growth hormone (GH), and linking anabolic processes to nutrient availability. Consideration of ligands and receptors, however, is insufficient to provide a complete understanding of the biology of IGF. This is because IGFs are found in binary complexes of 40-50 kDa with members of a family of IGF-binding proteins (IGFBPs-1 to -6) in all biological fluids. In addition, in postnatal serum, most IGFs are sequestered into ternary complexes of 150 kDa consisting of one molecule each of IGF, IGFBP-3 or IGFBP-5, and acid-labile subunit (ALS). Despite evidence that ALS plays an important role in the biology of circulating IGFs, it has received only limited attention relative to the other components of the IGF system. This review provides an overview on the current knowledge of ALS protein and gene structure, organisation and regulation by hormones, and insights from novel animal models such as the ALS knockout mice. PMID- 11431139 TI - Expression of metabotropic glutamate receptors in the rat and human testis. AB - The G protein-coupled receptor kinase type 4 mediates the homologous desensitisation of type-1 metabotropic glutamate (mGlu1) receptors and is predominantly expressed in the testis. Hence, we searched for the expression of mGlu1 or other mGlu receptor subtypes in rat and human testes. RT-PCR analysis showed the presence of mGlu1, -4 and -5 (but not -2 or -3) receptor mRNA in the rat testis. The presence of mGlu1 and -5 (but not mGlu2/3) receptor proteins was also demonstrated by Western blot analysis. In the rat testis, both mGlu1a and -5 receptors were highly expressed in cells of the germinal line. It is likely that these receptors are functional, because the agonist, (1S,3R)-1-aminocyclopentane 1,3-dicarboxylic acid, was able to stimulate inositol phospholipid hydrolysis in slices prepared from rat testes. Immunocytochemical analysis of bioptic samples from human testes showed a high expression of mGlu5 receptors inside the seminiferous tubuli, whereas mGlu1a immunoreactivity was restricted to intertubular spaces. mGlu5 receptors were also present in mature spermatozoa, where they were localised in the mid-piece and tail. This localisation coincided with that of beta-arrestin, a protein that is critically involved in the homologous desensitisation and internalisation of G protein-coupled receptors. Taken collectively, these results offer the first evidence for the expression of any glutamate receptor in testes, and suggest that at least mGlu5 receptors are present and functionally active in mature human sperm. PMID- 11431140 TI - Gonadal tumors of mice double transgenic for inhibin-alpha promoter-driven simian virus 40 T-antigen and herpes simplex virus thymidine kinase are sensitive to ganciclovir treatment. AB - We have previously produced transgenic (TG) mice expressing the mouse inhibin alpha-subunit promoter/Simian virus 40 T-antigen (Inhalpha/Tag) fusion gene. The mice develop gonadal somatic cell tumors at the age of 5-7 months; the ovarian tumors originate from granulosa cells, and those of the testes from Leydig cells. In the present study another TG mouse line was produced, expressing under the same inh-alpha promoter the herpes simplex virus thymidine kinase gene (Inhalpha/TK). Crossbreeding of the two TG mouse lines resulted in double TG mice (Inhalpha/TK-Inhalpha/Tag), which also developed gonadal tumors. The single (Inhalpha/Tag) and double TG (Inhalpha/TK-Inhalpha/Tag) mice, both bearing gonadal tumors, were treated at the age of 5.5-6.5 months with ganciclovir (GCV, 150 mg/kg body weight twice daily i.p.) for 14 days, or with aciclovir (ACV, 300 400 mg/kg body weight per day perorally) for 2 months. During GCV treatment, the total gonadal volume including the tumor, decreased in double TG mice by an average of 40% (P<0.05), while in single TG mice, there was a concomitant increase of 60% in gonadal size (P<0.05). GCV was also found to increase apoptosis in gonads of the double TG mice. Peroral treatment with ACV was less effective, it did not reduce significantly the gonadal volume. We also analyzed the in vitro efficacy of ACV and GCV treatments in transiently HSV-TK-transfected KK-1 murine granulosa tumor cells, originating from a single-positive Inhalpha/Tag mouse. GCV proved to be more effective and more specific than ACV in action. These results prove the principle that targeted expression of the HSV-TK gene in gonadal somatic cell tumors is potentially useful for tumor ablation by antiherpes treatment. The findings provide a lead for further development of somatic gene therapy for gonadal tumors. PMID- 11431141 TI - Differential regulation of the TRH gene promoter by triiodothyronine and dexamethasone in pancreatic islets. AB - TRH was initially found in the hypothalamus and regulates TSH secretion. TRH is also produced by insulin-containing beta-cells. Endogenous TRH positively regulates glucagon secretion and attenuates pancreatic exocrine secretion. We have previously shown that triiodothyronine (T(3)) down-regulates pre-pro-TRH gene expression in vivo and in vitro. The present study was designed to determine the initial impact of T(3) on rat TRH gene promoter and to compare this effect with that of dexamethasone (Dex). Primary islet cells and neoplastic cells (HIT T 15 and RIN m5F) were transiently transfected with fragments of the 5'-flanking sequence of TRH fused to the luciferase reporter gene. The persistence of high TRH concentrations in fetal islets in culture, probably due to transactivating factors, allowed us to explore how T(3) and Dex regulate the TRH promoter activity in transfected cells and whether the hormone effect is dependent on the cell type considered. TRH gene promoter activity is inhibited by T(3) in primary but not neoplastic cells and stimulated by Dex in both primary and neoplastic cells of islets. These findings validate previous in vivo and in vitro studies and indicate the transcriptional impact of these hormones on TRH gene expression in the pancreatic islets. PMID- 11431142 TI - Targeted disruption of the aromatase P450 gene (Cyp19) in mice and their ovarian and uterine responses to 17beta-oestradiol. AB - Aromatase P450 (CYP19) is an enzyme catalysing the conversion of androgens into oestrogens. We generated mice lacking aromatase activity (ArKO) by targeted disruption of Cyp19 and report the characteristic features of the ArKO ovaries and uteri as revealed by histological and biochemical analyses. ArKO females were totally infertile but there were as many developing follicles in their ovaries at 8 weeks of age as in wild-type ovaries. Nevertheless, no typical corpus luteum was observed in the ArKO ovaries. Electron microscopy revealed the presence of well-developed smooth endoplasmic reticulum, few lipid droplets and mitochondria with less organized tubular structures in the ArKO luteinized interstitial cells. These ultrastructural features were different from those of the wild-type interstitial cells, where there are many lipid droplets and mitochondria with well-developed tubular structures, characteristic of steroid-producing cells. When ArKO mice were supplemented with 17beta-oestradiol (E(2); 15 microg/mouse) every fourth day from 4 weeks of age for 1 month, increased numbers of follicles were observed in the ovaries as compared with those of untreated ArKO mice, although no typical corpus luteum was detectable. Ultrastructural analysis revealed the disappearance of the accumulated smooth endoplasmic reticulum in the luteinized interstitial cells after E(2 )supplementation. Transcripts of pro apoptotic genes such as p53 and Bax genes were markedly elevated in the ArKO ovaries as compared with those of wild-type mice. Although E(2) supplementation did not cause suppression of the elevated expression of p53 and Bax mRNAs, it caused marked enhancement of expression levels of lactoferrin and progesterone receptor mRNAs in the uteri as well as increases in uterine wet weight. At 8 months of age, ArKO mice developed haemorrhages in the ovaries, in which follicles were nearly depleted, while age-matched wild-type females still had many ovarian follicles. Furthermore, macrophage-like cells were occasionally observed in the ArKO ovarian follicles. These results suggested that targeted disruption of Cyp19 caused anovulation and precocious depletion of ovarian follicles. Additionally, analysis of mice supplemented with E(2) demonstrated that E(2) apparently supports development of ovarian follicles, although it did not restore the defect in ovulation. PMID- 11431143 TI - Correlation between inhibin secretion and damage of seminiferous tubules in a model of experimental autoimmune orchitis. AB - The aim of the present study was to evaluate inhibin secretion in rats with autoimmune orchitis. As we have previously described, experimental autoimmune orchitis (EAO) induced in rats by active immunization with testis homogenate and adjuvants is characterized by an interstitial mononuclear cell infiltrate and sloughing of the germinal epithelium. At 120 days after the first immunization 60% of the rats exhibited a severe orchitis with large areas of aspermatogenic seminiferous tubules in which only spermatogonia and Sertoli cells with cytoplasmic vacuolization remained attached to the tubular wall. None of the untreated (N) or control (C) rats revealed pathological alterations. Sixty percent decrease in testis weight was observed in rats with EAO compared with N or C groups. A 3-fold increase in serum FSH levels was observed in rats with EAO compared with N or C groups (19.8+/-3.7 vs 5.6+/-0.3 and 5.9+/-0.1 ng/ml respectively). A significant decrease in inhibin B levels was observed in rats with EAO when compared with N or C groups (40+/-4.6 vs 207+/-38.8 and 221.4+/ 28.6 pg/ml respectively). An inverse correlation between inhibin B and FSH serum levels and a direct correlation between inhibin B and testis weight were found. Strong expression of the inhibin alpha-subunit in Sertoli cells of untreated and control rats was observed; this subunit was undetectable or poorly detectable in rats with orchitis. Positive staining for the inhibin alpha-subunit was also observed in Leydig cells of all groups studied. In conclusion, using a model of autoimmune orchitis our results show that circulating inhibin B levels and inhibin alpha-subunit expression in Sertoli cell cytoplasm closely correlate with the degree of damage of the germinal epithelium. PMID- 11431144 TI - Interleukin-6 inhibits the expression of luteinizing hormone receptor mRNA during the maturation of cultured rat granulosa cells. AB - Interleukin-6 (IL-6) and its receptor components have been shown to be present in rat follicular granulosa cells. The present study was designed to examine the effect of this cytokine on changes in expression of the luteinizing hormone receptor (LHR) messenger RNA and of the steroidogenic enzyme, CYP11A1 (cytochrome P450 scc) in an in vitro model of granulosa cell maturation. Ovarian granulosa cells harvested from immature rats 2 days after treatment with equine chorionic gonadotropin were cultured for 48 h in media containing 10% fetal bovine serum. They were then transferred to a chemically defined serum-free medium and cultured for an additional 72 h. Within 24 h of transfer, the expressions of LHR and CYP11A1 mRNA increased significantly and remained increased for 72 h. The cells responded to exposure to FSH, but not LH, by an increase in production of cAMP before the additional 72 h of culture. The cAMP response to LH was attained within 24 h and persisted for 72 h, whereas the response to FSH decreased continuously with time. Inclusion of IL-6 in the culture medium caused a dose dependent decrease in expression of LHR mRNA, in addition to a decrease in the cAMP response to LH. Immunoneutralization of endogenous granulosa cell IL-6 resulted in an increase in expression of LHR mRNA, but not CYP11A1 mRNA. The results are consistent with the view that IL-6 may have a physiological role in the maturation of ovarian follicles by modulating the attainment of the LHR in granulosa cells. PMID- 11431145 TI - Feeding daidzein to late pregnant sows influences the estrogen receptor beta and type 1 insulin-like growth factor receptor mRNA expression in newborn piglets. AB - The present study was undertaken to determine the tissue-specific expression of estrogen receptor beta (ERbeta), and the effects of a daidzein supplement to the diet of pregnant sows on the expression of ERbeta, and type 1 insulin-like growth factor receptor (IGF-1R) genes in newborn piglets by using semi-quantitative RT PCR. Eight sows received a dietary supplement of daidzein at a dosage of 8 mg per kg feed from day 85 of gestation, and six sows were used as controls. After parturition, 2 male neonatal piglets were selected from each litter for sampling. ERbeta mRNA was detected in intestine, lung, thymus, kidney, pituitary and hypothalamus tissues, but not in heart, adrenal, skeletal muscle, liver or placental tissues. Daidzein treatment significantly increased the birth weight of male piglets and markedly reduced the level of ERbeta mRNA in the hypothalamus, but not in the pituitary. An up-regulation of IGF-1R gene transcription was observed in skeletal muscles of newborn piglets. In addition, the IGF-1R mRNA was found to be most abundant in pituitary and hypothalamus, followed by skeletal muscle, thymus, and liver tissues in decreasing order. Our results demonstrate that (1) ERbeta is expressed in a tissue-specific manner in newborn piglets, (2) daidzein down-regulates ERbeta gene expression in the hypothalamus, possibly indicating central effects of daidzein, and (3) daidzein influences fetal growth associated with higher IGF-IR gene expression in skeletal muscle. PMID- 11431146 TI - Interactions of synthetic estrogens with human estrogen receptors. AB - Synthetic estrogens have diverse chemical structures and may either positively or negatively affect the estrogenic signaling pathways through interactions with the estrogen receptors (ERs). Modeling studies suggest that 4-(1-adamantyl)phenol (AdP) and 4,4'-(1,3-adamantanediyl)diphenol (AdDP) can bind in the ligand binding site of ERalpha. We used fluorescence polarization (FP) to compare the binding affinities of AdP, AdDP and 2-(1-adamantyl)-4-methylphenol (AdMP) for human ERalpha and ERbeta with the binding affinities of the known ER ligands, diethylstilbestrol (DES) and 4hydroxytamoxifen (4OHT). Competition binding experiments show that AdDP has greater affinity for both ERs than does AdP, while AdMP does not bind the receptor proteins. The relative binding affinities of AdDP and AdP are weaker than the affinity of DES or 4OHT for both ERs with the exception of AdDP, which binds ERbeta with higher affinity than does 4OHT. We also found that AdDP and AdP cause differential conformational changes in ERalpha and ERbeta, which result in altered affinities of the ERs for fluorescein-labeled estrogen response elements (EREs) using a direct binding FP assay. The results show that ERbeta liganded with either AdDP or AdP has greater affinity for human pS2 ERE than the ERbeta-4OHT complex. The data suggest that synthetic molecules like adamantanes may function as biologically active ligands for human ERs. This demonstrates the importance of considering the potential of novel classes of synthetic compounds as selective ER modulators. PMID- 11431147 TI - Desensitization of beta2-adrenoceptor function in non-pregnant rat myometrium is modulated by sex steroids. AB - The effects of in vivo treatment with estrogen and progesterone on isoproterenol induced uterine relaxation and beta(2)-adrenoceptor (beta(2)AR) mRNA production in non-pregnant rat myometrium were investigated. Whether homologous myometrial desensitization of beta(2)AR function was dependent on or modulated by the two steroids was also examined. Estrogen treatment alone or in combination with progesterone reduced maximal relaxation (E(max)) of isolated uterine strips subsequently challenged with isoproterenol whereas progesterone alone had no effect on this parameter. The reduction was accompanied by an enhanced beta(2)AR mRNA concentration. The concentration of isoproterenol giving half-maximal relaxing response (EC(50)) increased following estrogen treatment and this effect was curbed by progesterone. Isoproterenol had no effect on beta(2)AR transcription irrespective of the steroid regimes employed. E(max) of isolated uterine strips was reduced following prolonged in vivo treatment with isoproterenol but the effect was found only when estrogen alone was administered concomitantly. Finally, in vivo treatment with isoproterenol increased EC(50) of uterine strips subsequently stimulated with isoproterenol in vitro. This effect was independent of steroid treatment. We conclude that homologous desensitization of beta(2)AR function in non-pregnant rat myometrium in terms of sensitivity (EC(50)) is independent of sex steroids but in terms of maximal response (E(max)) occurs only in the presence of estrogen. We speculate whether progesterone withdrawal in connection with the well-known estrogen dominance at rat parturition may strengthen the desensitization induced by beta(2)AR activation and thus contribute to the transformation of the uterus from a quiescent to a highly contractile organ. PMID- 11431148 TI - Urinary hormone levels during the natural menstrual cycle: the effect of age. AB - A number of studies have identified hormonal changes in women during their reproductive lifespan, many focusing upon changes in women over the age of 40 years. The present study has determined the effect of increasing age on hormone levels over three decades. Daily early morning urine samples were assayed for estrone-3-glucuronide (E3 G), pregnanediol-3-glucuronide (P3 G), testosterone-17 glucuronide (T17 G), FSH and LH. An examination of the validity of using creatinine as a volume adjuster in urine samples formed an integral part of the analysis. Volunteers were healthy women who had regular (25-35 days) cycles, were not taking oral contraceptives, hormone therapies or any other medication. Three age groups were compared: 20-29 years (n=13), 30-39 years (n=9) and 40-49 years (n=13). Statistical analyses were carried out using two-way ANOVA and post hoc t tests. Creatinine excretion, despite revealing no cycle-related variation in any age group, showed a decline with increasing age. Creatinine output was significantly lower in the 40-49 years age group in all phases of the cycle than in the 20-29 and 30-39 groups (P<0.0001). Uncorrected levels of E3 G were significantly higher in the 30-39 years group when compared with the 40-49 age group (P<0.0001). Uncorrected P3 G output was significantly higher in women aged 20-29 years than in women aged 40-49 years (P<0.001) and levels of uncorrected T17 G were higher in the 20-29 year age group when compared with the 30-39 or 40 49 years age group (P<0.0001). The present study is consistent with previous reports that have revealed a decline in creatinine clearance with increasing age, and therefore casts into some doubt the validity of using creatinine clearance as a procedure to correct for volume fluctuations in differing age groups of women. The study also demonstrates unequivocally that age-related variations in hormone levels are not restricted to women over 40 years of age. The novel finding of highly significant differences in mean levels of T17 G between the age groups is of considerable interest. It is presently unclear whether this resulted from specifically increased ovarian and/or adrenal secretion. The possible impacts of changes in testosterone levels during the female reproductive lifespan merits further study. PMID- 11431149 TI - Tissue-selective effects of continuous release of 2-hydroxyestrone and 16alpha hydroxyestrone on bone, uterus and mammary gland in ovariectomized growing rats. AB - 2-Hydroxyestrone (2-OHE(1)) and 16alpha-hydroxyestrone (16alpha-OHE(1)) have been reported to be risk factors for negative bone balance and breast cancer, respectively. The roles of these two metabolites of estrone as estrogen agonists or antagonists with respect to estrogen target tissues, or both, are poorly defined. The purpose of this study was to characterize metabolite and tissue specific differences between the actions of hydroxylated estrones on selected reproductive and non-reproductive estrogen target tissues in growing rats. First, the effects of ovariectomy were determined. Ovariectomy had the expected effects, including increases in all dynamic bone measurements at the proximal tibial epiphysis, without induction of bone loss. Second, ovariectomized growing rats were continuously treated for 3 weeks with 2-OHE(1), 16alpha-OHE(1), 17beta estradiol (E(2)), a combination of E(2) and 2-OHE(1) (E(2)+2-OHE(1)), or a combination of E(2) and 16alpha-OHE(1) (E(2)+16alpha-OHE(1)), using controlled release subcutaneous implanted pellets containing 5 mg 2-OHE(1), 5 mg 16alpha OHE(1), 0.05 mg E(2) or placebo. E(2) reduced body weight gain and radial and longitudinal bone growth as well as indices of cancellous bone turnover, and increased serum cholesterol, uterine wet weight and epithelial cell height, and proliferative cell nuclear antigen labeling in mammary gland. The hydroxylated estrones did not alter uterine wet weight and 16alpha-OHE(1) antagonized the E(2) stimulated increase in epithelial cell height. 2-OHE(1) had no effect on cortical bone, whereas 16alpha-OHE(1) was an estrogen agonist with respect to all cortical bone measurements. 16alpha-OHE(1) also behaved as an estrogen agonist with respect to serum cholesterol and cancellous bone measurements. 2-OHE(1) had no effect on most E(2)-regulated indices of cancellous bone growth and turnover, but was a weak estrogen agonist with respect to mineral apposition rate and bone formation rate. Neither estrogen metabolite influenced body weight gain. Third, weanling rats were treated for 1 week with vehicle, E(2) (200 microg/kg per day) or 16alpha-OHE(1) (30, 100, 300, 1000 and 3000 microg/kg per day) to confirm uterotropic effects of daily subcutaneous (s.c.) administration of 16alpha OHE(1). 16alpha-OHE(1) increased uterine weight in a dose-response manner to values that did not differ from rats treated with E(2). We conclude that the estrogen metabolites 2-OHE(1) and 16alpha-OHE(1) have target tissue-specific biological activities which differ from one another as well as from E(2). These findings add further support to the concept that there are several classes of estrogens with distinct biological activities. Furthermore, differences in the route of administration could influence the tissue specificity of estrogen metabolites. PMID- 11431150 TI - Osteoclast differentiation antigen, distinct from receptor activator of nuclear factor kappa B, is involved in osteoclastogenesis under calcitonin-regulated conditions. AB - Although calcitonin has been clinically utilized as a primary treatment for several metabolic bone diseases, its inhibitory effects against osteoclastic function diminish after several days owing to the calcitonin 'escape phenomenon'. We have previously found a unique cell-surface antigen (Kat1-antigen) expressed on rat osteoclasts. Here we show evidence that, in the presence of calcitonin, the Kat1-antigen is involved in osteoclastogenesis. Treatment of bone marrow cultures for forming osteoclast-like cells with anti-Kat1-antigen monoclonal antibody (mAb Kat1) provoked a marked stimulation of osteoclast-like cell formation only in the presence of calcitonin but not in its absence. Osteoclastogenesis stimulated by the receptor activator of nuclear factor kappa B (NF-kappaB) ligand/osteoclast differentiation factor was further augmented by mAb Kat1 in the presence of calcitonin. Furthermore, even in the presence of the osteoprotegerin/osteoclast inhibitory factor, mAb Kat1 induced osteoclast-like cell formation. Our current data suggest that the Kat1-antigen is a molecule that is distinct from receptor activator of NF-kappaB. The presence of the unique Kat1 antigen on cells in the osteoclast lineage appears to contribute to the fine regulation of osteoclastogenesis in vivo. Expression of this cell-surface molecule in cells in the osteoclast lineage may partly explain the mechanism responsible for the escape phenomenon. PMID- 11431151 TI - Cloning of the mouse sodium iodide symporter and its expression in the mammary gland and other tissues. AB - Iodide concentration in milk by mammals is a necessary step for thyroid hormone synthesis by the newborn. With the purpose of using the mouse as an animal model to analyse the role of the sodium iodide symporter (NIS) in iodide transport and its regulation in the mammary gland, mouse NIS (mNIS) cDNA was isolated from lactating mice. The cloned sequence shows an open reading frame of 1854 nucleotides encoding a protein of 618 amino acids highly homologous to the rat and human NIS (95% and 81% identity respectively). Expression of mNIS in cultured mammalian cells induced cellular iodide accumulation. This iodide uptake process is sodium dependent and inhibited by thiocyanate and perchlorate. Tissue distribution analysis revealed that mNIS mRNAs are predominantly expressed in thyroid, stomach and in the lactating mammary gland and are present to a lower extent in several other tissues. Our data show for the first time that the level of mNIS mRNA is upregulated in the mammary gland during lactation. PMID- 11431152 TI - In vitro free radical scavenging capacity of thyroid hormones and structural analogues. AB - It was reported that thyroid hormones decreased Cu(2+)-induced low-density lipoprotein (LDL) oxidation in vitro. Here, we investigated free radical scavenging capacities of thyroid hormones (3,5,3'-tri-iodo-L-thyronine (T(3)), thyroxine (T(4)) and 3,3',5'-tri-iodo-L-thyronine (rT(3))) and structural analogues (L-thyronine (T(0)), 3,5,3'tri-iodothyroacetic acid (TA(3)) and 3,5,3',5'-tetra-iodothyroacetic acid (TA(4))), using three different models of free radical generation. T(0), T(3) and TA(3) slowed down production of conjugated diene and thiobarbituric acid-reactive substances during LDL oxidation by 2,2'-azobis-[2-amidinopropane] (water-soluble), whereas rT(3), T(4) and TA(4) had practically no effect. In this system, T(0) was the more active compound. Using a 1,1-diphenyl-2-picrylhydrazyl (lipid-soluble) test, all compounds also revealed free radical scavenging capacities, but rT(3), T(4) and TA(4) were more active than T(0), T(3) and TA(3). T(3) was able to scavenge superoxide anion and hydroxyl radicals generated in an aqueous phase by a xanthine-xanthine oxidase system, as measured by electron paramagnetic resonance spectroscopy. It may be concluded that: (1) thyroid hormones and analogues with a 4'-hydroxy diphenylether structure have free radical scavenging capacities, (2) this property is influenced by the number of iodines on the phenolic ring, and (3) thyroid hormone scavenging capacity should not be the only mechanism explaining their protective effect on Cu(2+)-induced LDL oxidation. The physiological significance of the findings is discussed. PMID- 11431153 TI - Long-term thyroxine administration increases heat stress protein-70 mRNA expression and attenuates p38 MAP kinase activity in response to ischaemia. AB - The present study was undertaken to investigate heat stress protein (HSP)-70 mRNA induction and p38 MAP kinase (MAPK) activity in response to ischaemic stress in the hyperthyroid rat heart. L-Thyroxine (T(4)) (25 microg/100 g body weight) was administered to Wistar rats for 2 days (THYRacute) or 14 days (THYR), while animals treated similarly with normal saline served as controls (NORMacute and NORM). In addition, abdominal aortic banding was performed in another group of rats to produce constriction-induced hypertrophy (HYP), while sham-operated (SOP) animals served as controls. Isolated rat hearts were perfused in a Langendorff mode. Hearts from NORMacute (n=6), THYRacute animals (n=8), NORM (n=6), THYR (n=6), SOP (n=5) and HYP (n=7) animals were subjected to 20 min of zero-flow global ischaemia followed by 45 min of reperfusion. HSP70 mRNA expression and phosphorylated p38 MAPK protein expression were detected in response to ischaemia and protein kinase C-epsilon (PKCepsilon) protein expression was detected at baseline. Thyroid hormones were measured in plasma. Long-term T(4) administration and aortic constriction resulted in the development of cardiac hypertrophy. Thyroid hormones were increased in both THYR and THYRacute as compared with normal groups (P<0.05). HSP70 mRNA induction was increased 2.3-fold in THYR as compared with NORM hearts (P<0.05), whereas there was not any difference between THYRacute and NORMacute hearts (P>0.05). Phosphorylated p38 MAPK protein expression was 2.2-fold more in NORM than in THYR hearts (P<0.05), but it was not different between NORMacute and THYRacute hearts (P>0.05). HSP70 mRNA induction was 1.8-fold greater in HYP than in SOP hearts (P<0.05), whereas phosphorylated p38 MAPK protein expression was similar between the two groups (P>0.05). PKCepsilon protein expression at baseline was 1.7-fold more in NORM than in THYR hearts (P<0.05), and not different between NORMacute and THYRacute hearts (P>0.05) as well as HYP and SOP hearts (P>0.05). This study shows that HSP70 mRNA expression is increased, whereas p38 MAPK activation is attenuated in response to ischaemia in long-term T(4)-treated rat hearts as compared with normal and acute hyperthyroid hearts. PMID- 11431154 TI - Identification of thyroglobulin domain(s) involved in cell-surface binding and endocytosis. AB - Thyroglobulin (Tg) binds to cell surfaces through various binding sites of high, moderate and low affinity. We have previously shown that binding with low to moderate affinity is pH dependent, selective, but not tissue specific. To identify the regions of Tg involved in this cell surface binding, we studied the binding of (125)I-labeled cyanogen bromide peptides from human Tg to cell surfaces of thyroid cells (inside-out follicles) and of CHO cells. Electrophoretic analysis of cell homogenates after binding of native or of reduced and alkylated (125)I-labeled peptides showed that three peptides, P1, P2 and P3, were always associated with the cells. Sequence analysis allowed the identification of P1 (Ser-2445 to Met-2596 or Met-2610) and P2 (Phe-2156 to Met 2306). P3 proved to be a mixture of several peptides among which two were identified: P3-1 (Cys-1306 to Met-1640) and P3-2 (Cys-2035 to Met-2413) which includes P2. P1, P2 and P3-2 are entirely (P1) or partly (P2 and P3-2) located in the C-terminal domain of Tg homologous with acetylcholinesterase. The smallest peptides, P1 and P2, were purified by preparative electrophoresis. They both displayed strong binding properties towards cell surfaces. Inhibition experiments of (125)I-labeled Tg binding by P1 or P2 indicated that they were involved in Tg binding to cell surfaces. All the other peptides tested for their binding abilities were either not or only poorly involved in Tg binding to cell surfaces, which suggested that P1 and P2 are major Tg sites of binding to cell surfaces. These two peptides are not involved in the binding of Tg to the known Tg 'receptors' described in the literature, to which recycling, transcytosis and regulation functions have been ascribed. Thus they are potential tools to identify cell surface components involved in the process of Tg endocytosis leading to lysosomal degradation. PMID- 11431155 TI - Growth hormone endocrinology of Atlantic salmon (Salmo salar): pituitary gene expression, hormone storage, secretion and plasma levels during parr-smolt transformation. AB - A number of studies on the Atlantic salmon (Salmo salar), have reported changes in plasma GH during parr-smolt transformation, but there is a lack of information about the endocrinology of the GH system during this process. In order to elucidate the mechanisms underlying these changes in plasma GH levels during the parr-smolt transformation of Atlantic salmon, GH mRNA expression in the pituitary was studied together with total pituitary GH content, in vitro GH secretion rate and plasma GH and IGF-I levels. Atlantic salmon were kept in outside tanks, under natural condition from early February until late June. Approximately three times a month fish were killed and pituitaries and blood were sampled for investigation. Further, pituitaries were moved to the laboratory for in vitro GH secretion studies. The results show that the GH system is first activated by an increase in GH secretion rate, which leads to an increase in plasma GH levels and causes a drop in the total GH content of the pituitary. This drop in pituitary GH content is later reversed by an increased GH synthesis seen as an increase in GH mRNA expression. Maximal activation of the GH system is seen to occur in early May, when plasma IGF-I levels reach highest levels, after which a certain deactivation of the GH system takes place. The data show that plasma levels of GH are to a large extent regulated by the secretion rate from the pituitary, although changes in the GH clearance rate are also likely to take place and influence the plasma GH levels. The study further underlines the significant role that the GH-IGF-I axis plays in the parr-smolt transformation of the Atlantic salmon. PMID- 11431156 TI - GH-releasing peptide-6 overcomes refractoriness of somatotropes to GHRH after feeding. AB - After a meal, somatotropes are temporarily refractory to growth hormone-releasing hormone (GHRH), the principal hormone that stimulates secretion of growth hormone (GH). Refractoriness is particularly evident when free access to feed is restricted to a 2-h period each day. GH-releasing peptide-6 (GHRP-6), a synthetic peptide, also stimulates secretion of GH from somatotropes. Because GHRH and GHRP 6 act via different receptors, we hypothesized that GHRP-6 would increase GHRH induced secretion of GH after feeding. Initially, we determined that intravenous injection of GHRP-6 at 1, 3 and 10 microg/kg body weight (BW) stimulated secretion of GH in a dose-dependent manner. Next, we determined that GHRP-6- and GHRH-induced secretion of GH was lower 1 h after feeding (22.5 and 20 ng/ml respectively) than 1 h before feeding (53.5 and 64.5 ng/ml respectively; pooleds.e.m.=8.5). However, a combination of GHRP-6 at 3 microg/kg BW and GHRH at 0.2 microg/kg BW synergistically induced an equal and massive release of GH before and after feeding that was fivefold greater than GHRH-induced release of GH after feeding. Furthermore, the combination of GHRP-6 and GHRH synergistically increased release of GH from somatotropes cultured in vitro. However, it was not clear if GHRP-6 acted only on somatotropes or also acted at the hypothalamus. Therefore, we wanted to determine if GHRP-6 stimulated secretion of GHRH or inhibited secretion of somatostatin, or both. GHRP-6 stimulated secretion of GHRH from bovine hypothalamic slices, but did not alter secretion of somatostatin. We conclude that GHRP-6 acts at the hypothalamus to stimulate secretion of GHRH, and at somatotropes to restore and enhance the responsiveness of somatotropes to GHRH. PMID- 11431157 TI - Kallikrein gene expression in the gonadotrophin-stimulated rat ovary. AB - The kallikreins (KLKs) are a highly conserved multi-gene family of serine proteases that are expressed in a wide variety of tissues and act on a diverse range of substrates. KLK-like enzyme activity has variously been reported to increase or decrease during the period leading up to ovulation in the equine chorionic gonadotrophin (eCG)primed, human chorionic gonadotrophin (hCG) stimulated immature rat ovary. These earlier studies, which used biochemical assays to detect enzyme activity, lacked the specificity and sensitivity necessary to characterise conclusively the activity of the individual KLK gene family members. In this study, we have used a gene-specific RT-PCR/Southern hybridisation strategy to delineate the expression patterns of six of the individual KLK genes expressed in the rat ovary (rKLK1-3 and rKLK7-9). We have identified three broad patterns of expression in the eCG/hCG-stimulated ovary in which there is either a post-eCG increase/pre-ovulatory decrease in rKLK expression (rKLK1, rKLK3), a peri-ovulatory decrease in expression (rKLK2, rKLK8) or a relatively unchanged pattern of expression (rKLK7, rKLK9). In addition to clarifying the earlier biochemical studies, these findings support a differential role for the individual KLKs in the ovulatory process. PMID- 11431158 TI - Systemic administration of adrenomedullin(27-52) increases bone volume and strength in male mice. AB - Adrenomedullin is a 52-amino acid peptide first described in a human phaeochromocytoma but since been found to be present in many tissues, including the vascular system and bone. Because of its structural similarity to amylin and calcitonin gene-related peptide, both of which have actions on bone cells, we have previously assessed the effects of adrenomedullin on the skeleton, and found that it increases osteoblast proliferation in vitro and bone formation following local injection in vivo. The present study carries this work forward by assessing the effects on bone of the systemic administration of a fragment of this peptide lacking the structural requirements for vasodilator activity. Two groups of 20 adult male mice received 20 injections of human adrenomedullin(27-52) 8.1 microg or vehicle over a 4-week period and bone histomorphometry and strength were assessed. In the tibia, adrenomedullin(27-52) produced increases in the indices of osteoblast activity, osteoid perimeter and osteoblast perimeter (P<0.05 for both using Student's t-test). Osteoclast perimeter was not affected. There was a 21% increase in cortical width and a 45% increase in trabecular bone volume in animals treated with adrenomedullin(27-52) (P<0.002 for both). Assessment of bone strength by three-point bending of the humerus showed both the maximal force and the displacement to the point of failure were increased in the animals treated with adrenomedullin(27-52) (P<0.03 for both). There was also a significant increase in the thickness of the epiphyseal growth plate. No adverse effects of the treatment were noted. It is concluded that adrenomedullin(27-52) acts as an anabolic agent on bone. These findings may be relevant to the normal regulation of bone mass and to the design of agents for the treatment of osteoporosis. PMID- 11431159 TI - Autocrine role of adrenomedullin in the human adrenal cortex. AB - Previous studies from our laboratory have reported that adrenomedullin is synthesised in rat zona glomerulosa cells. In the present studies, it was found that the human adrenocortical cell line H295R expresses the gene encoding adrenomedullin, and that immunoreactive adrenomedullin is released into the culture medium. Furthermore, it was found that secretion of adrenomedullin is regulated by angiotensin II and forskolin. Studies on the actions of adrenomedullin and calcitonin gene-related peptide (CGRP) revealed a stimulatory effect of adrenomedullin, but not of CGRP, on aldosterone and cortisol secretion. These data suggest that adrenomedullin is not acting by a CGRP receptor-mediated mechanism in the H295R cell line. Adrenomedullin was also found to increase cAMP production, suggesting that in the adrenal, as in other cell types, cAMP is a second messenger for adrenomedullin action. However, the effects of adrenomedullin were not fully mimicked by forskolin, possibly suggesting a role for an additional second messenger. The presence of mRNA encoding both the putative adrenomedullin receptors, L1 and calcitonin receptorlike receptor/receptor-associated modulatory protein 2 (CRLR/RAMP-2), was demonstrated in H295R cells, but RAMP-1 was not detected, suggesting that these cells do not express the CGRPI receptor CRLR/RAMP-1. Taken together, these data have demonstrated that adrenomedullin is synthesised and secreted by H295R cells. The observed rate of adrenomedullin synthesis suggests that this peptide exerts a paracrine/autocrine effect in this adrenocortical cell line, probably acting through a specific adrenomedullin receptor, to stimulate steroidogenesis and increase aldosterone synthase expression. PMID- 11431160 TI - Uroguanylin and guanylate cyclase C in the human pancreas: expression and mutuality of ligand/receptor localization as indicators of intercellular paracrine signaling pathways. AB - The intestinal peptide hormone uroguanylin regulates electrolyte/fluid transport in the gastrointestinal epithelium by binding to its receptor, guanylate cyclase C (GC-C), and thus specifically coupling to activation of cystic fibrosis transmembrane conductance regulator (CFTR). Since CFTR is crucially involved in pancreatic electrolyte secretion, we investigated the human pancreas for expression and cell-specific localization of uroguanylin and guanylate cyclase C as potential regulatory components of pancreatic electrolyte secretion. RT-PCR analyses with specific primers revealed that uroguanylin and GC-C are expressed in the human pancreas (and in the duodenum, used as positive control); at the translational level, western blotting analyses with peptide- and region-specific antibodies identified the presence of 12.5 kDa uroguanylin and 130 kDa GC-C in both human pancreatic and intestinal extracts. At the cellular level, uroguanylin and GC-C immunoreactivities were absent from the islets of Langerhans but were exclusively confined to the exocrine parenchyma. Hence, uroguanylin was localized to the centroacinar cells typical of the pancreas, and also to epithelial cells of the intercalated, intralobular and interlobular ducts where the peptide was primarily concentrated adluminally to the apical portion of the respective cells. Coincidently, correlative studies localized the GC-C receptor to the epithelial cells of the ductal network, where it was confined exclusively to the apical cell membrane that evidently represents the functionally relevant target membrane domain for the regulatory peptide. In view of the fact that CFTR is highly expressed in pancreatic ductal cells where uroguanylin and its receptor are also localized, we assume that uroguanylin, an intrinsic pancreatic peptide, is involved in the regulation of electrolyte/water secretion in the ductal system via GC-C and CFTR. The particular cellular expression of uroguanylin in duct cells and the localization of GC-C to the duct cell apical membrane domain predict a novel route of intercellular signaling and luminal activation of GC-C via the pancreatic juice. PMID- 11431161 TI - Daily patterns of plasma leptin in sheep: effects of photoperiod and food intake. AB - Circulating concentrations of leptin in sheep correlate with body fatness and are affected by level of food intake and photoperiod. The present objective was to elucidate the short-term dynamics of leptin secretion. Frequent blood samples were taken over 48 h from 12 Soay rams after 16 weeks in short-day photoperiod (SD, 16 h darkness:8 h light) with freely available food, and then after 16 weeks in long days (16 h light:8 h darkness) with food freely available (LD) or restricted to 90% maintenance (LDR) (n=6/group). During the second 24 h of sampling, half were food deprived (n=6, SD and LD) and half had their meal times shifted (n=6, SD and LDR). A homologous RIA was developed, using antibodies raised in chicken against recombinant ovine leptin, to measure plasma concentrations. Simultaneous 24 h profiles of plasma insulin, glucose and non esterified fatty acids (NEFA) were measured. Plasma leptin was higher in LD than SD, and in LD than LDR, associated with higher food intake, liveweight and body condition score (adiposity), but tended to be lower in LDR than SD, associated with lower food intake, liveweight and body condition score. There was no evidence for a circadian rhythm of plasma leptin, but clear evidence for post prandial peaks of low amplitude (15-36%) 2-8 h after meals given at normal and shifted times. Complete food deprivation caused a dramatic fall in plasma leptin to basal levels within 24 h. There was a positive association of plasma leptin with plasma insulin, and negative association with NEFA, both between meals and during fasting. Thus, plasma leptin concentrations in sheep are sensitive to short-term changes in energy balance, as well as to long-term photoperiod-driven changes in food intake and adiposity. PMID- 11431162 TI - Androgen induces differentiation of a human papillomavirus 16 E6/E7 immortalized prostate epithelial cell line. AB - Androgen signaling is crucial for the growth and development, as well as for tumorigenesis of the prostate. However, many of the prostate epithelial cell lines developed previously, either normal or tumorigenic, do not express androgen receptor (AR) or respond to androgen. In order to advance our understanding on how androgen signaling regulates the growth and the differentiation status, and affects tumorigenicity of the epithelial cell, we performed experiments on HPr-1, a prostate cell line recently immortalized from normal human prostate epithelial cells. In the present study, AR was stably transfected into HPr-1 cells by replication-defective retrovirus. Treatment of HPr-1AR cells with androgen resulted in cell differentiation and growth retardation accompanied with up regulation of cytokeratins K8 and K18, prostate specific antigen, p21 and p27, and down-regulation of c-myc, bcl-2 and telomerase activity. Our results suggest that androgen promotes the process of differentiation in a human papillomavirus 16 E6/E7 immortalized prostate epithelial cell line which may reflect the normal effects of androgen on prostate cells. PMID- 11431163 TI - Chronic food-restriction alters the expression of somatostatin and growth hormone releasing hormone in the ovariectomised ewe. AB - Changes in the secretion of GH induced by long-term alterations in nutritional status are thought to result from alterations in somatostatin (SRIF) and growth hormone-releasing hormone (GHRH) at the level of the hypothalamus. To date however, the effect of nutrition on the gene expression of SRIF and GHRH in a species where GH secretion is increased by food restriction, as is the case for the sheep and human, remains unknown. We determined the effect of under-nutrition on the expression of SRIF and GHRH in the hypothalamus of sheep. Ovariectomised ewes were randomly divided into two groups and either fed an ad lib diet (n=6) or a restricted diet of 500 g lucerne chaff per day (food-restricted; n=5) for 7 months. In situ hybridisation was used to study hypothalamic gene expression for GHRH, SRIF and galanin (GAL). The food-restricted animals had elevated plasma concentrations of GH; this was associated with an increase in GHRH mRNA levels in the arcuate nucleus (ARC) and reduced SRIF in the rostral periventricular nucleus and ventromedial hypothalamic nucleus. The level of gene expression of GAL in the ARC and SRIF in the caudal periventricular nucleus was similar in ad lib and food restricted animals. In conclusion, the effect of chronic food-restriction on the secretion of GH reflects increased GHRH and reduced SRIF synthesis in the hypothalamus. PMID- 11431164 TI - Dreamy state: a case report from the selected writings of John Hughlings Jackson. Brain 1874. AB - This masterly account of a remarkable type of epileptic attacks with "dreamy state" was published in Brain, volume XI, pp. 179-207 in 1888 [1]. The patient was himself a physician and a highly articulate person endowed with an unusual capacity for introspection in addition to an interest in poetry and travelling 1. Jackson disagreed with some of the terms used by his doctor-patient; he preferred the word "reminiscence" to that of "recollection" and put between inverted commas that of "aura", indicating that he did not use it for any form of the "dreamy state" although the term remains easy to understand 2. Edited by James Taylor, Gordon Holmes and F.M.R. Walshe, Nijmegen: Arts and Boeve, 1996. PMID- 11431166 TI - A clinical spectrum of the myoclonic manifestations associated with typical absences in childhood absence epilepsy. A video-polygraphic study. AB - We investigated the electroclinical features of 12 patients with childhood absence epilepsy (CAE), presenting with typical absence seizures associated with myoclonic manifestations of the face or neck. All patients underwent repeated and prolonged split-screen video-polygraphic EEG recordings. The polygraphic recordings and clinical correlations of the absence seizures were analysed. All patients presented with multi-quotidian, typical absence seizures. During the absences, the patients could show mild, rhythmic, myoclonic jerks involving facial areas (eyebrows, nostrils, perioral region, chin) or neck muscles (sternocleidomastoideus), with the same frequency as the spike-wave complexes. Polygraphic tracings demonstrated that the myoclonias were correlated to the spike component. Clinically, all patients showed a benign course, with complete seizure control under antiepileptic treatment. In the follow-up, 7 patients withdrew from treatment without relapse. We conclude that all our patients showed an electroclinical picture consistent with CAE. The occurrence of myoclonic manifestations of the face or neck associated with the absences did not influence the benign course of their disease. The electroclinical features observed in our group of patients differentiates our cases both from epilepsy with myoclonic absences and from absences with perioral myoclonia (with Video). PMID- 11431167 TI - Idiopathic generalised epilepsy with phantom absences and absence status in a child. AB - A syndrome of idiopathic generalised epilepsy with phantom absences of undetermined onset has been recently described. This syndrome clinically becomes apparent in adulthood with generalised tonic clonic seizures and frequently absence status epilepticus. We report an 11 year-old normal girl with frequent episodes of absence status and no other overt clinical manifestations. However, appropriate video-EEG recordings documented that she had frequent absence seizures that were so mild as to escape recognition by her and the parents. These consisted of mild impairment of cognition and eyelid fluttering during brief generalised discharges of spike/multiple spike and slow waves. No further seizures occurred and the EEG normalised after appropriate drug treatment. Thus, it appears that this syndrome of phantom absences and absence status may start much earlier, in late childhood. Appropriate video-EEG documentation is needed for the recognition of these patients that may be more common than it appears from the few published cases (with Video). PMID- 11431168 TI - Spatio-temporal imaging of focal interictal epileptiform activity using EEG triggered functional MRI. AB - EEG-triggered, blood oxygen level-dependent functional MRI (BOLD-fMRI) was used in 24 patients with localization-related epilepsy and frequent interictal epileptiform discharges (spikes) to identify those brain areas involved in generating the spikes, and to study the evolution of the BOLD signal change over time. The location of the fMRI activation was compared with the scalp EEG spike focus and the structural MR abnormality. Twelve patients (50%) had an fMRI activation concordant with the EEG focus and structural brain abnormalities where present (n = 7). In 2 other patients, the fMRI activation was non-concordant with electroclinical findings. The remaining 10 patients (41.7%) showed no significant fMRI activation. These patients had significantly lower mean spike amplitudes compared to those with positive fMRI results (p = 0.03). The time course of the BOLD response was studied in 3 patients and this revealed a maximum signal change 1.5 to 7.5 sec after the spike. In conclusion, EEG-triggered fMRI can directly identify the generators of interictal epileptiform activity, with high spatial resolution, in selected patients with frequent spikes. The superior spatial resolution obtainable through EEG-triggered fMRI may provide an additional non invasive tool in the presurgical evaluation of patients with intractable focal seizures. PMID- 11431169 TI - Intraoperative methods for confirmation of correct placement of the vagus nerve stimulator. AB - Vagus nerve stimulation is a progressive therapy for intractable epilepsy. Variations in cervical anatomy can complicate localization of the vagus nerve and may lead to inappropriate placement of the stimulator leads. We have developed two intraoperative techniques that improve correct identification of the vagus nerve. Both of these techniques utilize the co-localization of the recurrent laryngeal nerve with the vagus nerve. For patients undergoing stimulator placement with regional and local anesthesia, the stimulator current intensity is increased until alteration of voice can be confirmed with a voice test. Patients undergoing general anesthesia can also be tested by direct stimulation of the isolated vagus nerve. Utilizing visualization of the larynx and vocal cords via fiberoptic endoscopy, direct stimulation of the vagus nerve will produce a contraction of the left lateral wall of the larynx and tightening of the left vocal cord. Neither of these procedures produce any untoward effects for the patients. We have found these methods improve our ability to confirm correct placement of the stimulator with minimal increase in operative time (with Video). PMID- 11431170 TI - Accessibility of spoken, written, and sign language in Landau-Kleffner syndrome: a linguistic and functional MRI study. AB - Landau-Kleffner syndrome (LKS) is an acquired aphasia which begins in childhood and is thought to arise from an epileptic disorder within the auditory speech cortex. Although the epilepsy usually subsides at puberty, a severe communication impairment often persists. Here we report on a detailed study of a 26-year old, left-handed male, with onset of LKS at age 5 years, who is aphasic for English but who learned British Sign Language (BSL) at age 13. We have investigated his skills in different language modalities, recorded EEGs during wakefulness, sleep, and under conditions of auditory stimulation, measured brain stem auditory-evoked potentials (BAEP), and performed functional MRI (fMRI) during a range of linguistic tasks. Our investigation demonstrated severe restrictions in comprehension and production of spoken English as well as lip-reading, while reading was comparatively less impaired. BSL was by far the most efficient mode of communication. All EEG recordings were normal, while BAEP showed minor abnormalities. fMRI revealed: 1) powerful and extensive bilateral (R > L) activation of auditory cortices in response to heard speech, much stronger than when listening to music; 2) very little response to silent lip-reading; 3) strong activation in the temporo-parieto-occipital association cortex, exclusively in the right hemisphere (RH), when viewing BSL signs. Analysis of these findings provides novel insights into the disturbance of the auditory speech cortex which underlies LKS and its diagnostic evaluation by fMRI, and underpins a strategy of restoring communication abilities in LKS through a natural sign language of the deaf (with Video) PMID- 11431171 TI - Comparison of twice- and three times daily tiagabine for the adjunctive treatment of partial seizures in refractory patients with epilepsy: an open label, randomised, parallel-group study. AB - A multicentre, open label, randomised, parallel group study compared the efficacy, tolerability and safety of two dosing regimens, t.i.d. and b.i.d., of tiagabine as an adjunctive therapy for the treatment of refractory patients with partial seizures. A total of 347 patients were randomised and treated (175 t.i.d. and 172 b.i.d.). Each group was administered the same daily dose of tiagabine incremented stepwise during a 12-week fixed-schedule titration period to a target dose of 40 mg/day. The patients were followed for a further 12-week flexible continuation phase. A significantly greater number of patients in the t.i.d. group completed the fixed schedule titration period (81.4% versus 73.1%; 95% CI of odds ratio = 0.331, 0.970; p = 0.038). The proportion of responders (patients showing at least a 50% decrease in all-seizure frequency from baseline) was similar for both groups (42.3% for b.i.d. and 47.1% for t.i.d.) during the last 8 weeks of the flexible phase and seven (4.0%) patients in the b.i.d. group were seizure-free compared to 14 (8.1%) patients in the t.i.d. group. Adverse events were of similar incidence in both groups, and mainly occurred during the fixed schedule titration period; they were mainly mild and CNS-related with somnolence being the most frequently reported. CONCLUSION: Tiagabine was effective as add-on treatment of refractory partial seizures. Although both regimens appear to offer a similar efficacy and safety profile, significantly more patients completed the study in the t.i.d. group compared to b.i.d., probably as a consequence of a lesser tolerability when high doses are given undivided. These results confirm that a slow titration and appropriate adjustment of dosing are essential conditions to ensure optimal use of tiagabine. PMID- 11431174 TI - Erythropoietin and transferrin metabolism in nephrotic syndrome. AB - Nephrotic syndrome is characterized by marked urinary excretion of albumin and other intermediate-size plasma proteins. This results in a profound alteration of the metabolism of many plasma proteins and protein-bound substances, as well as certain cellular and tissue proteins. This review summarizes available data on the effect of nephrotic syndrome on the metabolism and regulation of erythropoietin (EPO) and transferrin, which are essential for erythropoiesis. Studies of humans and animals have documented significant urinary losses of both EPO and transferrin in nephrotic syndrome. Urinary losses of EPO have been shown to cause EPO-deficiency anemia and prevent the normal increase in plasma EPO level in response to anemia and hypoxia in nephrotic syndrome. Similarly, transferrinuria and increased transferrin catabolism have been shown to cause hypotransferrinemia and, in some cases, iron-deficiency anemia. In addition, dissociation of iron from filtered transferrin, occasioned by a reduction in tubular fluid pH, can promote tubulointerstitial injury through the iron catalyzed generation of oxygen free radicals. This can account in part for the role of proteinuria as a risk factor for the progression of renal disease. Subcutaneous administration of recombinant EPO has been successfully used in the management of EPO-deficiency anemia in nephrotic syndrome. Similarly, iron supplementation and nutritional support are indicated in nephrotic patients with severe transferrinuria and iron-deficiency anemia. However, correction or amelioration of the underlying proteinuria, when possible, is the ideal approach to reversal of these complications. PMID- 11431175 TI - Contribution of gene polymorphisms in the renin-angiotensin system to macroangiopathy in patients with diabetic nephropathy. AB - The renin-angiotensin system is important in the control of hemodynamic status and pathogenesis of macrovascular disease, which is a major cause of morbidity and mortality in patients with type 2 diabetes with nephropathy. Serum angiotensin-converting enzyme (ACE) and angiotensinogen (Atg) levels are related to their respective gene polymorphisms. Seventy patients with type 2 diabetes with overt nephropathy (serum creatinine >/= 1.5 mg/dL) were studied. Serum ACE activity was measured by the spectrophotometric method. ACE deletion/insertion (D/I) and Atg M235T genotypes were determined by polymerase chain reaction. Patients with and without macroangiopathy were compared. Those with macroangiopathy had increased ACE activity (median, 60.9 U/L; range, 37.9 to 100 U/L versus without macroangiopathy, 47.9 U/L; range, 11.2 to 84.5 U/L; P = 0.01) and prevalence of ACE DD/DI genotypes (DD/DI:II: with macroangiopathy, 61%:39% versus without macroangiopathy, 34%:66%; P = 0.03). Multivariate analysis using age; sex; duration of diabetes; glycemic, blood pressure, and lipid level control; serum creatinine level; and presence of the ACE D allele showed that presence of the D allele (P = 0.03; odds ratio, 1.8; confidence interval, 1.1 to 3.1) and serum creatinine level (P = 0.0007) were independent risk factors for macroangiopathy. Association of the D allele became insignificant after serum ACE activity was included in the analysis in which only serum ACE activity (P = 0.004) and serum creatinine level (P = 0.01) were independent risk factors. Neither Atg M235T nor its synergistic effect with the ACE D allele showed an association with macroangiopathy. In conclusion, the ACE D allele is associated with macroangiopathy in Chinese patients with type 2 diabetes with nephropathy. The association is dependent on its effect on serum ACE activity, which is an independent risk factor for the development of macroangiopathy. PMID- 11431176 TI - Coadministration of losartan and enalapril exerts additive antiproteinuric effect in IgA nephropathy. AB - Angiotensin-converting enzyme (ACE) inhibitors and AT1-receptor antagonists (ARAs) are widely administered to reduce urinary protein loss and slow the progression of proteinuric nephropathy to end-stage renal failure. Our group recently observed that the combination of ACE inhibitors and ARAs may have an additive antiproteinuric effect, which may occur because ACE inhibitors do not completely reduce angiotensin II (Ang II) production. Ang II is also produced by chymase. Thus, combination therapy better antagonizes the effects of Ang II. The purpose of this study is to ascertain whether the additive antiproteinuric effect of ACE inhibitors plus ARAs is dose dependent and related to the drug-induced reduction in systemic blood pressure. Therefore, enalapril (E; 10 mg/d) and losartan (LOS; 50 mg/d) were randomly administered alone and then in association; initial dosages were doubled when drugs were administered alone and in association. To determine the influence of the drug-dependent effect on reducing blood pressure and the reduction in urinary proteinuria, both ambulatory and office blood pressures were recorded. E and LOS administered alone reduced proteinuria by the same extent; no further reduction was observed when E and LOS alone were administered at a doubled dose. When E and LOS were coadministered, proteinuria decreased by a greater extent compared with E and LOS alone; an additional reduction in proteinuria was observed when combined therapy doses were doubled. The reduction in proteinuria was not correlated with clinical through blood pressure; however, reductions in diastolic and mean ambulatory blood pressures significantly correlated with the decrease in proteinuria, as well as with creatinine clearance. In conclusion, this study shows that combination therapy with E and LOS has an additive dose-dependent antiproteinuric effect that is likely induced by the drug-related reduction in systemic blood pressure. In normotensive proteinuric patients, it is likely that even a small reduction in systemic blood pressure may affect intraglomerular hemodynamics by a great extent because efferent arteriole regulation is hampered more completely by the coadministration of ACE inhibitors and ARAs. PMID- 11431177 TI - Renal lesions in leprosy: a retrospective study of 199 autopsies. AB - In the present work, 199 patients with leprosy who underwent autopsy between 1970 and 1986 were retrospectively studied to determine the prevalence, types, clinical characteristics, and etiologic factors of renal lesions (RLs) in leprosy. Patients were divided into two groups: 144 patients with RLs (RL+) and 55 patients without RLs (RL-). RLs observed in 72% of the autopsied patients were amyloidosis (AMY) in 61 patients (31%), glomerulonephritis (GN) in 29 patients (14%), nephrosclerosis (NPS) in 22 patients (11%), tubulointerstitial nephritis (TIN) in 18 patients (9%), granuloma in 2 patients (1%), and other lesions in 12 patients (6%). AMY occurred most frequently in patients with lepromatous leprosy (36%; nonlepromatous leprosy, 5%; P < 0.01), recurrent erythema nodosum leprosum (33%; P < 0.02), and trophic ulcers (27%; 0.05 < P < 0.10). Ninety-seven percent of AMY was found in patients with lepromatous leprosy, 88% showed recurrent trophic ulcers, and 76% presented with erythema nodosum leprosum. NPS was found in older patients with arterial hypertension, neoplastic diseases, infectious diseases, and vasculitis associated with GN. Most patients with AMY presented with proteinuria (95%) and renal failure (88%). The most frequent causes of death were renal failure in patients with AMY (57%), infectious diseases in patients with GN (41%) and TIN (45%), and cardiovascular diseases in patients with NPS (41%). No difference in survival rates was observed among RL- patients and those with AMY, GN, NPS, or TIN. PMID- 11431178 TI - What happens to renal transplant recipients who lose their grafts? AB - Little attention has been given to the fate of patients who lose their grafts. We reviewed outcomes of 438 recipients of first renal allografts who underwent transplantation at our institution between January 1, 1988, and December 31, 1997, and lost their grafts or died with a functioning transplant. Of the 438 patients, 168 patients died with a functioning transplant. The most common causes of death were cardiac disease, infection, and cancer. Patients who died with a functioning graft were older (>49 years, 64.3%) than patients who died after returning to dialysis therapy or who are still alive (>49 years, 25.9%). Eighty six patients (39%) who returned to dialysis therapy were again placed on a cadaveric waiting list. Only 44 patients received a second transplant, of which 30 transplants (68.2%) are still functioning. Our study shows that relatively few patients who lose kidney transplants are returned to the cadaveric waiting list and even fewer undergo retransplantation. PMID- 11431179 TI - Nonreferral and nonacceptance to dialysis by primary care physicians and nephrologists in Canada and the United States. AB - Research from Canada and the United States suggests that not offering dialysis to patients who might benefit still occurs. This study was conducted to investigate nonreferral and nonacceptance to dialysis by primary care physicians (PCPs) and nephrologists in these countries. We surveyed a random sample of Canadian and US PCPs and nephrologists concerning their attitudes toward and experience with withholding dialysis in patients with advanced chronic renal failure. In response to a question about whether the physician believes there should be an age beyond which dialysis should not be offered, 12% of Canadian PCPs, 20% of US PCPs, 4% of Canadian nephrologists, and 9% of US nephrologists answered yes. When asked about their recommendations concerning dialysis initiation in 10 vignettes of patients with impending end-stage renal disease (ESRD), the responses of Canadian and US physicians were similar. PCPs compared with nephrologists were less likely to recommend dialysis in cases with physical illnesses and more likely to recommend it in cases with neuropsychiatric impairments. Over a 3-year period, 13% of Canadian PCPs and 19% of US PCPs reported nonreferral to dialysis at least once. Withholding rates were 25% for Canadian PCPs, 16% for US PCPs, 13% for Canadian nephrologists, and 17% for US nephrologists. We conclude that although nonreferral of patients who might benefit from dialysis still occurs, it does not seem to be common, and the attitudes of Canadian and US physicians toward this issue are similar and could not entirely account for the much greater incidence of treated ESRD in the United States. PCPs and nephrologists should continue to be educated about the modern criteria for patient selection for dialysis. PMID- 11431180 TI - End-stage renal disease: factors affecting referral decisions by family physicians in Canada, the United States, and Britain. AB - The objective of this study is to determine how patient age, sex, creatinine level, and comorbidity affect referral decisions for the treatment of end-stage renal disease (ESRD) and whether these decisions are affected by physician characteristics in three countries: Canada, the United States, and Britain. A vignette-based questionnaire was mailed to a random sample of family physicians in Ontario, Canada (1,818 physicians); all family physicians in the state of New York (1,814 physicians); and a sample of general practitioners from the south of England (2,228 physicians) in 1996. Physicians were presented with clinical scenarios involving a patient with varying degrees of renal insufficiency and a complicating comorbidity, including angina, diabetes, cancer, mental illness, or socioeconomic circumstances. They were asked to indicate the likelihood of referral. Half the physicians received a questionnaire describing a male patient, and half, a female patient. Mean creatinine levels at which physicians would refer were 260 micromol/L for British physicians, 297 micromol/L for Canadian physicians, and 340 micromol/L for American physicians. No difference in referral rates was found based on the sex of the patient or physician. Sixty-five percent of American and Canadian physicians would refer regardless of patient age, but only 49% of British physicians would do so. Family physicians in the United States, Canada, and Britain function as gatekeepers for patients with ESRD. They are less likely to refer based on increasing severity of comorbid conditions. They also discriminate based on age, but not sex. PMID- 11431181 TI - Recreational drug use: a neglected risk factor for end-stage renal disease. AB - Case series have suggested that heroin and cocaine users are at increased risk for renal failure, but the contribution of heroin and other addictive drugs to the incidence of end-stage renal disease (ESRD) in the general population remains unknown. To clarify this issue, we conducted a case-control study in the general population to examine associations between drug use and treated ESRD. Cases were 716 patients who started therapy for ESRD in 1991, identified through a regional registry. Controls were 361 persons of similar age (20 to 65 years) selected by random digit dialing. Main risk factors examined were the lifetime use of heroin, cocaine, and other addictive drugs, assessed by telephone interview. After adjustment for age, sex, race, socioeconomic status, and history of hypertension and diabetes, persons who had ever used heroin or other opiates (any amount) were at increased risk for ESRD (adjusted odds ratio, 19.1; 95% confidence interval, 1.7 to 208.7). After adjustment for the same sociodemographic and medical history variables, the use of cocaine or crack and psychedelic drugs was also associated with ESRD, but these associations could not be separated from the effects of heroin use. PMID- 11431182 TI - Long-term outcomes of revascularization for peripheral vascular disease in end stage renal disease patients. AB - The occurrence of peripheral vascular disease (PVD) and atraumatic lower extremity amputations is significantly greater in patients with end-stage renal disease (ESRD) than those with normal renal function. Moreover, the mortality for dialysis patients undergoing atraumatic lower-extremity amputations is far greater. Because PVD requiring amputation is an extreme form of PVD, we tested the hypothesis that mortality and intermediate outcomes for patients with ESRD undergoing lower-extremity revascularization, a less extreme form of PVD, would be equivalent to that for patients without ESRD. This is a retrospective case control analysis of lower-extremity revascularization in patients with ESRD. Procedures in patients with ESRD were matched with procedures in non-ESRD controls for patient age, sex, race, diabetes mellitus, and hospital setting. Patient survival, graft survival, and limb salvage rates were determined using Kaplan-Meier analysis. Subjective interpretation of functional and symptomatic improvement was determined by telephone interviews with patients or relatives. Thirty-one procedures were performed on 20 patients with ESRD and 64 matched procedures were performed on 57 patients without ESRD. In the ESRD group, median patient survival was 1.72 years compared with 5.17 years for the control group (P < 0.001). Time to 50% limb loss was 1.24 years in the ESRD group and longer than 5.65 years in the control group (P < 0.001). Time to 50% graft patency loss was 0.70 years in the ESRD group and longer than 5.5 years in the control group (P < 0.05). Subjective improvement was less in patients with ESRD. Outcomes of lower extremity revascularization in patients with ESRD are inferior to those in non ESRD controls. The mortality rate for patients with ESRD who undergo revascularization is extremely high. Patient-related variables (eg, increased prevalence of hypertension and cardiovascular disease) and/or provider-specific factors (eg, timing of surgery in the course of PVD) may be responsible for poorer outcomes. PMID- 11431183 TI - Significant association between the progression of coronary artery calcification and dyslipidemia in patients on chronic hemodialysis. AB - Quantification of coronary artery calcification (CAC) determined by electron-beam computed tomography (EBCT), known as the CAC score (CACS), correlates with total plaque amount and coronary risk factors and strongly associates with maximal stenosis in the epicardial arteries. However, data are limited concerning the CACS in chronic dialysis patients, although atherosclerotic vascular disease is the most frequent complication. We examined the relation between coronary risk factors, metabolic factors of calcium and other minerals, and CACS progression in 24 dialysis patients. The mean patient age was 53 +/- 14 (SD) years, and mean duration of dialysis was 64 +/- 69 months. In each patient, the CACS was measured twice, with a mean interscan period of 17 +/- 3 months. The mean CACS progressed from 449 +/- 605 to 669 +/- 894 overall, and the mean change in CACS (DeltaCACS) was 220 +/- 78. Patients were divided into two groups: slow progressors, with DeltaCACS of 7.5 +/- 31 (n = 12), and rapid progressors, with DeltaCACS of 432 +/ 458 (n = 12). Triglyceride concentrations (198 +/- 65 versus 103 +/- 50 mg/dL; P < 0.001) were high, and high-density lipoprotein cholesterol (HDL-C) concentrations (46 +/- 11 versus 60 +/- 18 mg/dL; P < 0.05) were low in rapid progressors. Rapid progression of CAC was associated with high triglyceride and low HDL-C concentrations. The clinical significance of these observations remains to be determined. PMID- 11431184 TI - Cholesteryl ester transfer protein as a protective factor against vascular disease in hemodialysis patients. AB - Cholesteryl ester transfer protein (CETP) may promote reverse cholesterol transport. An elevated concentration of high-density lipoprotein cholesterol (HDL C) is a protective factor against atherosclerosis. However, the effects of CETP itself and its interaction with HDL-C have not been investigated in hemodialysis patients, who are at high risk for atherosclerosis and generally considered to have decreased reverse cholesterol transport. We investigated the independent or synergistic influence of postprandial serum CETP and HDL-C concentrations on apparent atherosclerotic complications in 202 hemodialysis patients aged 40 to 80 years. Patients with vascular disease (n = 39) had significantly lower concentrations of CETP than those without vascular disease (n = 163). When all study subjects were divided into four groups according to CETP and HDL-C concentrations based on median values, we found significant differences in the prevalence of vascular disease (test for trend, P < 0.005): 28.8% in group I (HDL C < 48 mg/dL; CETP < 2.2 microgram/mL); 25.0% in group II (HDL-C < 48 mg/dL; CETP >/= 2.2 microgram/mL); 17.8% in group III (HDL-C >/= 48 mg/dL; CETP < 2.2 microgram/mL); and 6.9% in group IV (HDL-C >/= 48 mg/dL; CETP >/= 2.2 microgram/mL). Multiple logistic regression analysis retained the interaction term between HDL-C (in milligrams per deciliter) and CETP (in micrograms per milliliter), but not HDL-C or CETP itself, as inversely associated with vascular disease in the entire patient group. In patients with HDL-C levels at the median value or greater, CETP had an independent odds ratio of 0.31 (95% confidence interval, 0.1 to 0.97; P < 0.05) after adjusting for age. These results suggest that CETP may serve as a protective factor against vascular disease in hemodialysis patients, especially those with normal or elevated HDL-C concentrations. PMID- 11431185 TI - Association of two MTHFR polymorphisms with total homocysteine plasma levels in dialysis patients. AB - The effect of the combined 5,10-methylenetetrahydrofolate reductase (MTHFR) 677C- >T and 1298A-->C genotype on total homocysteine (tHcy), folate, and vitamin B(12) plasma levels was investigated in 983 subjects, including 415 hemodialysis patients, 179 peritoneal dialysis patients, and 389 healthy individuals. Mean tHcy plasma concentrations were 27.2 +/- 15.8 micromol/L in hemodialysis patients, 25.4 +/- 19.1 micromol/L in peritoneal dialysis patients, and 8.9 +/- 3.5 micromol/L in healthy individuals. Hyperhomocysteinemia (tHcy > 15 micromol/L) was detected in 81.6% of patients and 2.6% of controls. Multiple stepwise regression analysis showed that the MTHFR 677C-->T/1298A-->C genotype (CC/AA, CC/AC, CC/CC, CT/AA, CT/AC, TT/AA), vitamin use, age, folate and vitamin B(12) plasma level were significant predictors of tHcy plasma levels. Analysis of variance showed that this effect of MTHFR genotypes on tHcy level was caused by significantly greater tHcy levels in 677TT/1298AA hemodialysis and peritoneal dialysis patients versus other genotypes. Compound heterozygous controls (677CT/1298AC genotype) had significantly greater tHcy levels compared with 677CC/1298AA controls. There was no major effect of MTHFR polymorphisms on folate and vitamin B(12) plasma concentrations. This study shows that the MTHFR 677TT/1298AA genotype, but not the 677CT/1298AC genotype, is a significant predictor of tHcy plasma levels in dialysis patients. PMID- 11431186 TI - Residual renal function and mortality risk in hemodialysis patients. AB - Residual renal function, defined as the urinary clearance of urea and creatinine, is minimal in many patients treated with hemodialysis (HD) and tends to be ignored in most outcome studies involving HD patients. Recent studies showed that residual renal function, even at a low level, is influential in preventing mortality in the minority of patients with end-stage renal disease treated with peritoneal dialysis. This issue generally has not been examined in patients treated with HD. This prospective observational study of all 114 patients at a single community-based freestanding HD center is designed to examine the impact of residual renal function (defined as renal urea clearance and renal creatinine clearance derived from 24-hour urinary volumes) on mortality over a 2-year period. During that period, 50 deaths occurred in 114 patients. The presence of residual renal function was protective against mortality (odds ratio for death, 0.44; 95% confidence interval, 0.24 to 0.81; P = 0.008), even after adjustment for duration of dialysis treatment, age, smoking, presence of diabetes, presence of cardiovascular disease, serum albumin level, and urea reduction rate. In conclusion, the presence of residual renal function, even at a low level, is associated with a lower mortality risk in HD patients. PMID- 11431187 TI - Hepatitis C screening strategies in hemodialysis patients. AB - Hepatitis C virus (HCV) infection is common in patients undergoing chronic hemodialysis, with an estimated yearly incidence of 0.2% and prevalence between 8% and 10%. Although a screening strategy based on alanine aminotransferase (ALT) values is currently recommended, this strategy has not been evaluated for cost effectiveness compared with other potential screening strategies. A comparison therefore was made using a decision-analysis model of a simulated cohort of 5,000 hemodialysis patients followed up for 5 years. Using direct medical costs, three strategies were evaluated, including: (1) ALT values with confirmatory testing (biochemical), (2) serial enzyme-linked immunosorbent and strip immunoblot assay testing (serological), and (3) polymerase chain reaction (viral). Under baseline assumptions, the per-patient cost of screening hemodialysis patients for HCV was $378 for biochemical-based testing, $195 for serological-based testing, and $696 for viral-based testing. Our model was robust when varying the costs of testing, as well as the incidence and prevalence of HCV infection. Results of sensitivity analysis by varying costs, HCV incidence, and HCV prevalence indicated that serological-based screening was less costly than biochemical testing. Biochemical testing was in turn less costly than viral-based screening. Serological-based testing was also more effective in the diagnosis of de novo HCV infection, with a likelihood ratio of 85, in contrast to the likelihood ratio of 44 with biochemical-based testing using viral-based screening as the gold standard. A serological-based screening strategy is less costly and more effective than biochemical-based screening in the diagnosis of de novo HCV infection. Serological-based screening should be considered for HCV screening in hemodialysis populations. PMID- 11431188 TI - Noninvasive stool antigen assay can effectively screen Helicobacter pylori Infection and assess success of eradication therapy in hemodialysis patients. AB - Helicobacter pylori (H pylori) stool antigen (HpSA), serological antibody against H pylori (immunoglobulin G [IgG]), and urea breath test (UBT) are noninvasive methods used to detect H pylori infection that can allow a patient to avoid the discomfort and risk of invasive endoscopy. However, because the UBT has proven not highly reliable in patients with end-stage renal disease (ESRD), this study attempts to investigate the diagnostic efficacy of HpSA and IgG for H pylori detection in 80 patients with ESRD and 80 dyspeptic patients without renal function impairment as a control group. All patients in both study groups underwent panendoscopy to obtain gastric biopsy specimens for histological examination and H pylori culture. With H pylori infection defined as a positive result on either histological examination or culture, we evaluated the reliability of HpSA and serum IgG in detecting H pylori infection. Forty of the patients with ESRD (50%) and 48 patients in the control group (60%) were proven to be infected with H pylori. To eradicate H pylori infection, these patients were administered a 1-week course of triple therapy. To evaluate the success of H pylori eradication, 38 patients in the ESRD group and 44 patients in the control group underwent a follow-up endoscopy and provided stool samples for HpSA 6 to 8 weeks later. Success of H pylori eradication was found in 86.8% of the patients with ESRD (33 of 38 patents) and 84.1% of the control patients (37 of 44 patients). Before therapy, HpSA for H pylori detection was 97.5% sensitive and 97.5% specific in patients with ESRD, as effective as that in the control group. After therapy, HpSA was 100% sensitive and more than 96% specific to detect the failure of H pylori eradication therapy in both the ESRD and control groups. Conversely, the use of IgG as a screening method for H pylori infection proved to be less effective because it showed a sensitivity of 87.5% and specificity of 80% in this study. Monitoring the success of triple therapy, IgG had a specificity of only 21.9% in the ESRD group and 24.3% in the control group. In summary, HpSA is a noninvasive and reliable tool to screen H pylori infection before therapy and assess the success of eradication therapy in patients with ESRD. PMID- 11431189 TI - A crossover study of gabapentin in treatment of restless legs syndrome among hemodialysis patients. AB - Restless legs syndrome (RLS) is a common entity affecting hemodialysis patients. Although the cause of RLS remains unclear, a number of therapies have been used successfully to treat the disorder. Gabapentin is an anticonvulsant shown to alleviate symptoms of RLS in two small studies of nonhemodialysis patients. Because it is excreted renally, gabapentin has a long half-life among hemodialysis patients and may be advantageous if proven effective. We conducted a randomized, double-blind, placebo-crossover study of gabapentin in the treatment of RLS among a population of hemodialysis patients. Sixteen patients identified with RLS were randomized to either gabapentin or placebo. After 6 weeks of treatment with 200 to 300 mg of gabapentin after each hemodialysis session, patients' RLS was reassessed. After a 1-week washout period, patients were switched from gabapentin to placebo or placebo to gabapentin. After another 6 weeks, patients' RLS was assessed again. Patient data were analyzed using both parametric and nonparametric means. Thirteen of the 16 original patients completed the study. Two patients dropped out because of lethargy (believed to be secondary to gabapentin), and 1 patient died secondary to myocardial infarction. Eleven patients responded to gabapentin, but not placebo (P < 0.01). One patient responded to both, and 1 patient responded to placebo, but not gabapentin. Gabapentin is an effective treatment for RLS in hemodialysis patients. PMID- 11431190 TI - Optimizing erythropoietin therapy in hemodialysis patients. AB - The European Best Practice Guidelines for the management of anemia in patients with chronic renal failure recommend the percentage of hypochromic red blood cells (%HRCs) as the best measure of iron use by erythropoietic tissues. They suggest that "sufficient iron should be administered to attain: serum ferritin 100 ng/mL, HRCs <10%. In practice, to achieve these minimum criteria will mean aiming for optimal levels of serum ferritin 200-500 ng/mL, HRCs <2.5%." We increased prospectively the delivered dose of iron supplements to a large (n = 228) unselected hemodialysis cohort with a sustained (24-month) hemoglobin (Hb) outcome meeting the UK Renal Association minimum standard of 85%, greater than or equal to 10.0 g/dL. This was managed through a computer-aided decision support system for erythropoietin (EPO) and intravenous iron sucrose therapy. Hb outcome was maintained with medians between 11.3 and 11.8 g/dL. Median red blood cell hypochromia (%HRCs) decreased from 8% (interquartile range [IQR], 3 to 15) to 4% (IQR, 2 to 8; P < 0.001, U-Mann Whitney test). Serum ferritin level increased from a median of 188 (IQR, 115 to 256) to 480 ng/mL (IQR, 397 to 595; P < 0.001, U-Mann Whitney test). Median EPO dose decreased from 136 (IQR, 83 to 216) to 72 IU/kg/wk (IQR, 33 to 134), which strongly correlated with median %HRCs through the range less than 10% (Spearman's correlation, 0.73; P < 0.01). These data suggest that EPO responsiveness continues to improve toward the normal range for %HRCs (<2.5%) and aspiring to values much less than 10% is cost-effective. The ferritin outcome required to achieve these lower values for %HRC outcome is greater than the current recommended range, although in steady state, the mean iron treatment dose is similar to that in previous studies (ie, approximately 60 mg/wk). PMID- 11431191 TI - A preceding exchange with polyglucose versus glucose solution modifies peritoneal equilibration test results. AB - The peritoneal equilibration test (PET) is an important tool for evaluating peritoneal membrane characteristics. The polyglucose icodextrin induces ultrafiltration caused by colloid osmosis through the small pores of the peritoneal membrane and therefore is especially effective during long dwell times. The main indications for polyglucose solutions are daytime dwells in patients on automated peritoneal dialysis and nighttime exchanges in continuous ambulatory peritoneal dialysis (CAPD) patients. In CAPD patients, PET is started immediately after the icodextrin exchange. Therefore, we performed two PETs in each of 15 CAPD patients. PET post-1.36% glucose was performed immediately after a preceding exchange with 2 L of 1.36% glucose dialysate solution (dwell time, 10 hours). PET postpolyglucose was started immediately after a preceding exchange with 2 L of 7.5% icodextrin solution (dwell time, 10 hours). The dialysate to plasma (D/P) ratio of creatinine, phosphate, and sodium during PET postpolyglucose was significantly greater than during PET post-1.36% glucose at 1, 2, 3, and 4 hours of dwell time. The quotient of dialysate glucose at 1, 2, and 4 hours of dwell time to dialysate glucose at 0 dwell time was significantly lower in PET postpolyglucose compared with PET post-1.36% glucose. In the case of creatinine, phosphate, and glucose, PET postpolyglucose curves tended to be steeper than those of PET post-1.36% glucose during the first hour of dwell time, whereas both curves were parallel between 1 and 4 hours of dwell time. The course of D/P ratio curves of urea nitrogen, protein, and albumin was nearly identical between PET postpolyglucose and PET post-1.36% glucose. In a subgroup of 5 patients, D/P ratios of creatinine and phosphate were also greater in PET postpolyglucose compared with PET performed after a long dwell with 2.27% glucose solution. Before a scheduled PET, CAPD patients using icodextrin solution during the nighttime should perform their nighttime exchange with conventional glucose solution. PMID- 11431192 TI - Cefepime versus vancomycin plus netilmicin therapy for continuous ambulatory peritoneal dialysis-associated peritonitis. AB - Cefepime is a cephalosporin with a broad spectrum of activity against most gram positive and gram-negative pathogens. In this study, we attempted to compare the safety and efficacy of cefepime monotherapy against the potentially more toxic combination of vancomycin and netilmicin in the treatment of continuous ambulatory peritoneal dialysis (CAPD)-associated bacterial peritonitis. Eighty one consecutive CAPD patients who presented with peritonitis from January 1, 1998, to June 30, 2000, were recruited for study. Patients were randomized to be administered either intraperitoneal (IP) cefepime, 1 g once daily (group A), or intravenous vancomycin and netilmicin at conventional doses (group B) for 10 days. Bacterial growth was obtained in 52 episodes (66%), and pathogens identified included gram-positive organisms (30 episodes; 38%), gram-negative organisms (14 episodes; 18%), mixed organisms (2 episodes; 2.5%), and fungus (6 episodes; 8%). Eight patients were excluded after randomization for various reasons (6 patients, fungal peritonitis; 2 patients, wrong diagnoses). Because of the relatively low peritonitis rate after the use of a disconnect system, the sample size of this study was relatively small, giving a power of 0.45. There were no significant differences in primary response rates and cure rates (no relapse >28 days after completion of antibiotic therapy) between both groups of patients (group A versus group B, 82% [32 of 39 patients] versus 85% [29 of 34 patients] and 72% [28 of 39 patients] versus 76% [26 of 34 patients], respectively; P = not significant). No significant side effect was encountered in either group. Total peritonitis-related hospitalizations were 84 patient-days (1, 7, 8, 11, 20, and 37 patient-days) and 115 patient-days (3, 6, 9, 14, 21, 21, and 41 patient-days), whereas total costs per patient cure were estimated to be US $1,039 and US $1,371 in groups A and B, respectively. We conclude that once-daily 1-g IP cefepime monotherapy is a simple, safe, and cost-effective alternative to vancomycin and netilmicin therapy in the treatment of CAPD-associated bacterial peritonitis. PMID- 11431193 TI - Long-term outcomes in simultaneous kidney-pancreas transplant recipients with portal-enteric versus systemic-bladder drainage. AB - We retrospectively reviewed long-term outcomes in simultaneous kidney-pancreas transplant (SKPT) recipients with portal-enteric (P-E) versus systemic-bladder (S B) drainage. Forty-five patients were alive with functioning grafts 1 year after SKPT and were followed up for a minimum of 3 years (mean, 5.9 years), including 26 patients with P-E drainage and 19 patients with S-B drainage. Recipient demographic and transplant characteristics were similar between the two groups. In both groups, hospital admissions decreased significantly with increasing time after SKPT, although significantly fewer readmissions occurred in the first year in the P-E than the S-B group. The most common reason for readmission in both groups was infection, followed by miscellaneous, surgical, and immunologic morbidity. The incidence of readmission for dehydration was significantly less in the P-E group (P < 0.01). Mean systolic and diastolic blood pressures were similar between groups, although the number of antihypertensive medications was significantly less in the S-B group. Although fasting C-peptide levels were significantly greater in the S-B group, the two groups were similar with regard to carbohydrate (fasting serum glucose, hemoglobin A(1c)) and lipid (total cholesterol) metabolism. Renal and pancreas allograft functions were similar between the two groups. At 1 year post-SKPT, stabilization in most diabetic complications was reported. Four quality-of-life surveys that provided 29 scores were completed 6 to 24 months (mean, 18.5 months) after SKPT. Improved quality of life was reported in all but one of the scales, with many dimensions showing significant improvements. At 3 years after SKPT, no activity limitation was reported in 76% of patients with P-E drainage versus 53% with S-B drainage (P = 0.11). Five-year actual patient, kidney, and pancreas graft survival rates after P-E versus S-B drainage are 92% and 84%, 81% and 79%, and 88% and 74%, respectively (P = not significant). SKPT with P-E drainage is a safe and effective method to treat advanced diabetic nephropathy and is associated with decreasing morbidity, improving rehabilitation and quality of life, and stablizing metabolic function over time. The long-term prognosis after the first year is excellent and at least similar to the results achieved with S-B drainage. PMID- 11431194 TI - Immuno-unreactive albumin excretion increases in streptozotocin diabetic rats. AB - We previously showed that albumin is fragmented (>90%) during renal passage to low-molecular-weight (<10 kd) peptides. The aim of the present study was to document the renal handling of albumin in experimental diabetes. Tritium-labeled albumin was infused into control and streptozotocin (STZ) diabetic rats during 7 days. Urinary radioactivity, assessed by size exclusion chromatography, revealed a major peak corresponding to low-molecular-weight, albumin-derived fragments and a minor peak corresponding to intact albumin or high-molecular-weight, albumin derived protein. The fractional clearance of albumin, calculated from total radioactivity measurements, was at least 100-fold greater than the fractional clearance of albumin determined by radioimmunoassay (RIA) for control and diabetic rats. This result was mainly because low-molecular-weight, albumin derived fragments were not detected by RIA. The fractional clearance of high molecular-weight, albumin-derived protein was 2- to 10-fold greater than the fractional clearance determined by RIA. The immuno-unreactive high-molecular weight, albumin-derived protein (called ghost albumin), characterized by size exclusion chromatography and high-performance liquid chromatography, was present in control and diabetic rat urine. Ghost albumin excretion rate was enhanced 11 fold after 8 weeks of STZ diabetes as compared with aged-matched controls. This study shows that renal modification resulting in low-molecular-weight and high molecular-weight components of albumin is a major contributor to the renal handling of albumin. The results indicate that excretion of modified albumin is increased in STZ rats as compared with albumin detected by conventional RIA. Long term studies are necessary to evaluate the potential of ghost albumin as a new marker for the assessment of urinary albumin in diabetes. PMID- 11431195 TI - Glomerular osteopontin expression and macrophage infiltration in glomerulosclerosis of DOCA-salt rats. AB - Expression of the chemoattractant osteopontin (OPN) may contribute to macrophage infiltration in many types of tubulointerstitial kidney disease, but the role of OPN in chronic glomerulosclerosis is unknown. We hypothesized that glomerular OPN expression and macrophage infiltration occur in deoxycorticosterone acetate (DOCA)-salt glomerulosclerosis in rats. Uninephrectomized rats receiving DOCA pellets and 1% saline were compared with control rats. OPN mRNA was determined by Northern blot, and OPN protein was determined by Western blot. The localization of OPN was studied by in situ hybridization and double immunohistochemistry with glomerular cell markers. Macrophage infiltration was quantified by counting ED-1 positive cells, and semiquantitative glomerulosclerosis scores were obtained. In DOCA-salt rats, OPN mRNA in the kidney was increased 2-fold over control after 9 days and 3 weeks and 20-fold after 6 weeks. Tubulointerstitial OPN staining was apparent after 21 days of DOCA treatment. Glomerular OPN mRNA and protein was detected after 42 days in parietal and visceral epithelial cells, activated myofibroblasts, and occasionally mesangial cells. Progressive glomerular macrophage infiltration occurred during the development of DOCA hypertension, paralleling the degree of glomerulosclerosis. Glomeruli staining positive for osteopontin contained more macrophages (18.4 +/- 3.4 per cross-section) than osteopontin-negative glomeruli (3.6 +/- 0.5; P < 0.05). Glomerular OPN expression occurs in chronic hypertensive glomerulosclerosis and is associated with macrophage infiltration. The data suggest a role for OPN as a chemoattractant in hypertensive glomerulosclerosis. PMID- 11431196 TI - Chronic renal failure, end-stage renal disease, and peritoneal dialysis in Gitelman's syndrome. AB - The chronic state of hypovolemia, hypotension, and hypokalemia found in Bartter's syndrome has been shown to lead to a chronic nephropathy, which then can progress toward end-stage renal disease and dialysis. This progression, however, has never been reported for Gitelman's syndrome, a variant of Bartter's syndrome that shows a milder clinical picture. This report is the first to document this progression (ie, the development of end-stage renal disease in Gitelman's syndrome) as well as the first report of the use of peritoneal dialysis in either Bartter's syndrome or Gitelman's syndrome. The clinical course highlights the importance of and the need for careful control of hemodynamic status in these patients to slow the progression of renal injury. The hemodynamic alterations that characterize Bartter's syndrome and Gitelman's syndrome patients suggest that for patients requiring renal replacement therapy, peritoneal dialysis is a more appropriate treatment because of its less severe impact on these parameters. PMID- 11431197 TI - Ranitidine-induced acute interstitial nephritis in a cadaveric renal allograft. AB - Ranitidine frequently is used for preventing peptic ulceration after renal transplantation. This drug occasionally has been associated with acute interstitial nephritis in native kidneys. There are no similar reports with renal transplantation. We report a case of ranitidine-induced acute interstitial nephritis in a recipient of a cadaveric renal allograft presenting with acute allograft dysfunction within 48 hours of exposure to the drug. The biopsy specimen showed pathognomonic features, including eosinophilic infiltration of the interstitial compartment. Allograft function improved rapidly and returned to baseline after stopping the drug. PMID- 11431198 TI - AN-69 membrane reactions are pH-dependent and preventable. AB - We report two pediatric patients who required blood priming for continuous venovenous hemodiafiltration. Both of these patients developed a significant hypotensive episode with initiation of continuous venovenous hemodiafiltration with immediate resolution on discontinuation. The most notable common characteristics of these patients were the use of the Multi-flo 60 (AN-69) dialyzer membrane and blood priming. No similar episodes were encountered when patients were primed with saline or albumin. The AN-69 membrane is exquisitely pH sensitive. The lower the pH concentration of the blood passing by the membrane, the greater the activation of bradykinin, a known hypotensive-inducing agent, by the dialyzer. On review of blood available from our blood bank, the following parameters became apparent. The pH of standard blood available from our blood bank ranged from 6.1 to 6.4. The blood obtained from our blood bank had significant hyperkalemia, hyponatremia, and hypocalcemia. No reactions were noted when patients were primed with normal saline, which has a pH of around 5.9. We speculate that the presence of endogenous blood substances, such as bradykinin, may have induced the hypotensive episodes. We describe two techniques we developed that should allow for the increased safe and effective use of the AN-69 membranes in continuous venovenous hemodiafiltration circuits. These observations indicate the requirement for careful and close attention to detail when delivering renal replacement therapy to anyone, but especially patients weighing less than 10 kg. PMID- 11431199 TI - Nephrology grows older and wiser on the backs of giants. PMID- 11431200 TI - IgA nephropathy and inhibitors of the renin angiotensin system: is reduction in proteinuria adequate proof of efficacy? PMID- 11431201 TI - Hepatitis C infection screening in hemodialysis units. PMID- 11431202 TI - Ethical issues in living organ donation: donor autonomy and beyond. AB - Despite nearly 50 years of experience with living kidney donation, ethical questions about this practice continue to haunt us today. In this editorial I will address two of them: (1) Given the possibility of limited understanding and coercion, how can we be sure that a person who offers to donate an organ is acting autonomously? and (2) Do people have a right to donate? The universal requirement for informed consent is the traditional method for ensuring that a person is acting autonomously. But, while obtaining fully informed consent is desirable, it may not always be achievable or necessary. When the recipient is very dear to the potential donor, the donor may base his decision primarily on care and concern rather than on a careful weighing of risks and benefits. I will argue that consent that emanates from such deep affection should be considered just as valid as consent that is fully informed. But consent is not enough. There is no absolute right to donate an organ. If there were such a right, then some physician would be obligated to remove an offered organ upon request, regardless of the risks involved. I do not believe that physicians have such an obligation. Physicians are moral agents who are responsible for their actions and for the welfare of their patients. Therefore, while the values and goals of the potential donor should be given great weight during the decision-making process, physicians may justifiably refuse to participate in living organ donation when they believe that the risks for the donor outweigh the benefits. PMID- 11431203 TI - Novel mechanism(s) implicated in tubular albumin reabsorption and handling. PMID- 11431204 TI - Acute renal failure in a 53-year-old woman with Crohn's disease treated with 5 aminosalicylic acid. PMID- 11431205 TI - Calculation and public use of the unit-specific standardized mortality ratio. PMID- 11431206 TI - Calculation and public use of the unit-specific standardized mortality ratio. PMID- 11431207 TI - Calculation and public use of the unit-specific standardized mortality ratio. PMID- 11431208 TI - Calculation and public use of the unit-specific standardized mortality ratio. PMID- 11431209 TI - Hematopoietic cell transplantation-related nephropathy. PMID- 11431210 TI - Biocompatibility of sorbent suspension dialysis in cirrhotic patients with hepatic encephalopathy. PMID- 11431211 TI - Estimating lean body mass by dual-energy x-ray absorptiometry. PMID- 11431212 TI - Is kidney transplantation lifesaving? PMID- 11431213 TI - Low parathyroid hormone levels and higher risk of hip fractures and mortality. PMID- 11431216 TI - Rapid recurrence of ANCA-negative pauci-immune vasculitis after cadaveric renal transplantation. AB - We report a patient with renal failure secondary to antineutrophil cytoplasmic autoantibody (ANCA)-negative pauci-immune systemic vasculitis who developed rapid recurrence of vasculitis involving skin, eye, and kidney allograft 1 month after cadaveric renal transplant while being maintained on cyclosporine, mycophenolate mofetil, and prednisolone. Despite early institution of aggressive treatment with plasma exchange, the disease remained uncontrolled and resulted in substantial morbidity and subsequent graft failure within 1 year post-transplantation. This case confirms that ANCA-negative vasculitides may recur in transplanted patients taking mycophenolate mofetil. Transplantation is contraindicated when active disease is evident. PMID- 11431217 TI - Diversity and challenges of child psychiatry. PMID- 11431218 TI - A psychiatric dialogue on the mind-body problem. AB - Of all the human professions, psychiatry is most centrally concerned with the relationship of mind and brain. In many clinical interactions, psychiatrists need to consider both subjective mental experiences and objective aspects of brain function. This article attempts to summarize, in the form of a dialogue between a philosophically informed attending psychiatrist and three residents, the major philosophical positions on the mind-body problem. The positions reviewed include the following: substance dualism, property dualism, type identity, token identity, functionalism, eliminative materialism, and explanatory dualism. This essay seeks to provide a brief user-friendly introduction, from a psychiatric perspective, to current thinking about the mind-body problem. PMID- 11431219 TI - The use of psychotropic medications during breast-feeding. AB - OBJECTIVE: The authors reviewed the risks and benefits regarding the use of psychiatric medications during breast-feeding as they relate to the health and well-being of mothers and their infants. Strategies are discussed to limit infant exposure to a medication while effectively treating the nursing mother. METHOD: A MEDLINE search of the literature since 1955 was conducted to determine the use of psychotropic medications in breast-feeding women. Search items included each of the categories of psychopharmacologic agents as well as each of the agents in association with nursing, breast-feeding, postpartum, lactation, and breast milk. RESULTS: No controlled studies on the safety of psychotropic medications in nursing mothers were found. Case reports and small case series for each of the different psychotropic medications serve as the basis for suggested treatment guidelines for the management of psychiatric illnesses in breast-feeding women. Thus, each case needs to be considered on an individual basis, with a thoughtful analysis of the risks and benefits of nursing and exposure of the infant to medication. The baseline clinical status of the infant should also be reviewed. CONCLUSIONS: Women are vulnerable postpartum to psychiatric disorders and frequently face the need to decide whether to take psychotropic medications while breast-feeding. Should psychiatric medication be indicated, the parents should be provided with the available information regarding the effects of these medications on the neonate. In this way, an informed decision can be made. When psychotropic medication is used during breast-feeding, it is strongly recommended that the infant's pediatrician be involved in monitoring the infant. PMID- 11431220 TI - Psychiatrist-patient boundary issues following treatment termination. AB - OBJECTIVE: Psychiatrist-patient relationships after termination of treatment are fraught with complexities and are the subject of ongoing debate. The authors discuss the issue of boundary violation allegations that arise after treatment has ended, with the goal of explicating how these issues have been handled in psychiatric discussions as well as in broader sociolegal settings. METHOD: Clinical illustrations and legal cases are used to illustrate how legal and administrative bodies have dealt with posttermination boundary issues. RESULTS: Courts and regulatory bodies have tended to use the psychoanalytic concept of transference to decide issues in which there has been a complaint of impropriety be it romantic, financial, or social in nature-arising after termination of treatment. However, a multitude of treatment approaches are currently employed in psychiatry, and often their practitioners either do not use the concept of transference or deny its validity. If the concept is used, it is often present in many settings outside therapy. CONCLUSIONS: The concept of transference is subject to continuing debate and modification within psychoanalysis, and its use in judicial or quasijudicial settings raises questions about whether it meets standards of scientific acceptance. Using the concept of transference to decide posttermination issues results not only in confusion but also has the potential for many adverse consequences for practitioners and the profession. PMID- 11431222 TI - Mothers' functioning and children's symptoms 5 years after a SCUD missile attack. AB - OBJECTIVE: The authors assessed the long-term consequences of the SCUD missile attack in Israel on children as a function of their mothers' psychological functioning, family cohesion, and the event itself. METHOD: Five years after the Gulf War, the authors assessed the internalizing, externalizing, stress, and posttraumatic symptoms of 81 children aged 8-10 years whose homes were damaged in the SCUD missile attack, as well as general and posttraumatic symptoms, defensive style, and object relations in their mothers. RESULTS: There was a significant decrease in severity in most symptom domains and an increase in avoidant symptoms in the children. Greater severity of symptoms was associated with being displaced, living in a family with inadequate cohesion, and having a mother with poor psychological functioning. The association between the symptoms of children and mothers was stronger among the younger children. Posttraumatic symptoms increased in one-third of the children and decreased in one-third over the last 30 months of the study. Severe posttraumatic symptoms were reported in 8% of the children. CONCLUSIONS: Despite a continuous decrease in symptom severity, risk factors identified shortly after the Gulf War continued to exert their influence on children 5 years after the traumatic exposure. PMID- 11431223 TI - The role of childhood interpersonal trauma in depersonalization disorder. AB - OBJECTIVE: In contrast to trauma's relationship with the other dissociative disorders, the relationship of trauma to depersonalization disorder is unknown. The purpose of this study was to systematically investigate the role of childhood interpersonal trauma in depersonalization disorder. METHOD: Forty-nine subjects with DSM-IV depersonalization disorder and 26 healthy comparison subjects who were free of lifetime axis I and II disorders and of comparable age and gender were administered the Dissociative Experiences Scale and the Childhood Trauma Interview, which measures separation or loss, physical neglect, emotional abuse, physical abuse, witnessing of violence, and sexual abuse. RESULTS: Childhood interpersonal trauma as a whole was highly predictive of both a diagnosis of depersonalization disorder and of scores denoting dissociation, pathological dissociation, and depersonalization. Emotional abuse, both in total score and in maximum severity, emerged as the most significant predictor both of a diagnosis of depersonalization disorder and of scores denoting depersonalization but not of general dissociation scores, which were better predicted by combined emotional and sexual abuse. The majority of the perpetrators of emotional abuse were either or both parents. Although different types of trauma were modestly correlated, only a few of these relationships were statistically significant, underscoring the importance of comprehensively considering different types of trauma in research studies. CONCLUSIONS: Childhood interpersonal trauma and, in particular, emotional abuse may play a role in the pathogenesis of depersonalization disorder. Compared to other types of childhood trauma, emotional maltreatment is a relatively neglected entity in psychiatric research and merits more attention. PMID- 11431224 TI - Dissociation as a mediator of psychopathology among sexually abused children and adolescents. AB - OBJECTIVE: This study investigated the role of dissociation as a mediator of mental health outcomes in children with a history of sexual abuse. METHOD: The study group consisted of 114 children and adolescents (ages 10-18 years) who were wards of the Illinois Department of Children and Family Services and were living in residential treatment centers. Interviews, provider ratings, and chart reviews were used to assess the relationship of childhood abuse history, dissociative responses, and psychopathology. RESULTS: Sexual abuse history was significantly associated with dissociation, whereas a history of physical abuse was not. Both sexual abuse and dissociation were independently associated with several indicators of mental health disturbance, including risk-taking behavior (suicidality, self-mutilation, and sexual aggression). Severity of sexual abuse was not associated with dissociation or psychopathology. Analysis of covariance indicated that dissociation had an important mediating role between sexual abuse and psychiatric disturbance. These results were replicated across several assessment sources and varied perspectives. CONCLUSIONS: The findings suggest a unique relationship between sexual abuse and dissociation. Dissociation may be a critical mediator of psychiatric symptoms and risk-taking behavior among sexually abused children. The assessment of dissociation among children may be an important aspect of treatment. PMID- 11431225 TI - Functional neuroanatomy of visuospatial working memory in fragile X syndrome: relation to behavioral and molecular measures. AB - OBJECTIVE: Fragile X syndrome is a neurogenetic disorder that is the most common known heritable cause of neurodevelopmental disability. This study examined the neural substrates of working memory in female subjects with fragile X syndrome. Possible correlations among behavioral measures, brain activation, and the FMR1 gene product (FMRP expression), as well as between IQ and behavioral measures, were investigated. METHOD: Functional magnetic resonance imaging was used to examine visuospatial working memory in 10 female subjects with fragile X syndrome and 15 typically developing female subjects (ages 10-23 years). Subjects performed standard 1-back and 2-back visuospatial working memory tasks. Brain activation was examined in four regions of the cortex known to play a critical role in visuospatial working memory. Correlations between behavioral, neuroimaging, and molecular measures were examined. RESULTS: Relative to the comparison group, subjects with fragile X syndrome performed significantly worse on the 2-back task but not on the 1-back task. In a region-of-interest analysis focused on the inferior frontal gyrus, middle frontal gyrus, superior parietal lobule, and supramarginal gyrus, comparison subjects showed significantly increased brain activation between the 1-back and 2-back tasks, but subjects with fragile X syndrome showed no change in activation between the two tasks. Significant correlations were found in comparison subjects between activation in the frontal and parietal regions and the rate of correct responses on the 2-back task, but not on the 1-back task. In subjects with fragile X syndrome, significant correlations were found during the 2-back task between FMRP expression and activation in the right inferior and bilateral middle frontal gyri and the bilateral supramarginal gyri. CONCLUSIONS: Subjects with fragile X syndrome are unable to modulate activation in the prefrontal and parietal cortex in response to an increasing working memory load, and these deficits are related to a lower level of FMRP expression in fragile X syndrome subjects than in normal comparison subjects. The observed correlations between biological markers and brain activation provide new evidence for links between gene expression and cognition. PMID- 11431226 TI - Meta-analysis of the association between the 7-repeat allele of the dopamine D(4) receptor gene and attention deficit hyperactivity disorder. AB - OBJECTIVE: Family, twin, and adoption studies show attention deficit hyperactivity disorder (ADHD) to have a substantial genetic component. Although several studies have shown an association between ADHD and the 7-repeat allele of the dopamine D(4) receptor gene (DRD4), several studies have not. Thus, the status of the ADHD-DRD4 association is uncertain. METHOD: Meta-analysis was applied to case-control and family-based studies of the association between ADHD and DRD4 to assess the joint evidence for the association, the influence of individual studies, and evidence for publication bias. RESULTS: For both the case control and family-based studies, the authors found 1) support for the association between ADHD and DRD4, 2) no evidence that this association was accounted for by any one study, and 3) no evidence for publication bias. CONCLUSIONS: Although the association between ADHD and DRD4 is small, these results suggest that it is real. Further studies are needed to clarify what variant of DRD4 (or some nearby gene) accounts for this association. PMID- 11431227 TI - Cholinergic activity in autism: abnormalities in the cerebral cortex and basal forebrain. AB - OBJECTIVE: Measures of cholinergic transmitter activity were investigated in patients with autism because of reported neuropathological abnormalities in cholinergic nuclei in the basal forebrain. METHOD: Levels of cholinergic enzyme and receptor activity were measured in the frontal and parietal cerebral cortex of deceased autistic adults, similarly aged normal adults without mental retardation, and nonautistic mentally retarded adults. The immunoreactivity levels of brain-derived neurotrophic factor and nerve growth factor were measured in the basal forebrain. RESULTS: There were no differences between the autistic and comparison groups in choline acetyltransferase or acetylcholinesterase activity in the cerebral cortex and basal forebrain or in muscarinic M(2) receptor or alpha-bungarotoxin binding within the cortex. Cortical M(1) receptor binding was up to 30% lower than normal in the autistic subjects, and the difference reached significance in the parietal cortex. In both the parietal and frontal cortices, differences in nicotinic receptors assessed by [(3)H]epibatidine binding were significant and extensive (65%-73% lower in the autistic group than in the normal subjects); there were no differences in nicotine binding in the basal forebrain. Immunochemical analysis indicated lower levels of both the alpha(4) and beta(2) nicotinic receptor subunits in the parietal cortex. The M(1) receptor abnormality was not evident in the nonautistic group with mental retardation, although the lower [(3)H]epibatidine binding was apparent. In the basal forebrain, the level of brain-derived neurotrophic factor in the autistic group was three times as high as the level of the normal group. CONCLUSIONS: These neurochemical abnormalities implicate the cholinergic system in developmental disorders such as autism and suggest the potential for intervention based on cholinergic receptor modulation. PMID- 11431228 TI - A placebo-controlled study of guanfacine in the treatment of children with tic disorders and attention deficit hyperactivity disorder. AB - OBJECTIVE: This study evaluated the efficacy and safety of guanfacine in treating children with tic disorders and attention deficit hyperactivity disorder (ADHD). METHOD: Subjects from a specialty tic disorders clinic were randomly assigned to receive 8 weeks of treatment with guanfacine or placebo under double-blind conditions. Follow-up visits occurred every 2 weeks for safety monitoring and dose adjustment. RESULTS: Thirty-four medication-free subjects (31 boys and three girls with a mean age of 10.4 years) with ADHD, combined type, and a tic disorder participated. After 8 weeks of treatment, guanfacine was associated with a mean improvement of 37% in the total score on the teacher-rated ADHD Rating Scale, compared to 8% improvement for placebo. Nine of 17 subjects who received guanfacine were blindly rated on the Clinical Global Improvement scale as either much improved or very much improved, compared with none of 17 subjects who received placebo. The mean score on the parent-rated hyperactivity index improved by 27% in the guanfacine group and 21% in the placebo group, not a significant difference. On the Continuous Performance Test, commission errors decreased by 22% and omission errors by 17% in the guanfacine group, compared with increases of 29% in commission errors and of 31% in omission errors in the placebo group. Tic severity decreased by 31% in the guanfacine group, compared to 0% in the placebo group. One guanfacine subject with sedation withdrew at week 4. Guanfacine was associated with insignificant decreases in blood pressure and pulse. CONCLUSIONS: Guanfacine appears to be a safe and effective treatment for children with tic disorders and ADHD. PMID- 11431229 TI - Subcortical correlates of craving in recently abstinent alcoholic patients. AB - OBJECTIVE: This study investigated functional cerebral correlates of craving in alcoholic patients and examined the state/trait characteristics of the regional cerebral network implicated in craving. METHOD: Functional magnetic resonance imaging (fMRI) was used to map cerebral response elicited by ethanol odor in 10 male patients with alcohol dependence who had undergone detoxification and 10 matched nonpatients. After 3 weeks, during which the patients underwent standardized behavioral therapy with psychopharmacological intervention, all subjects were studied a second time with fMRI to evaluate the effects of therapy on the functional cerebral correlates of craving. RESULTS: In the alcoholic patients, cue-induced craving before treatment elicited activation primarily in the subcortical-limbic region of the right amygdala/hippocampal area and in the cerebellum. After treatment, activation was found in the superior temporal sulcus, while subcortical or cerebellar participation was no longer present. Comparison subjects showed no comparable amygdala or cerebellar activation during ethanol stimulation and demonstrated no change in activation pattern between measurements. CONCLUSIONS: This investigation points to state-dependent neurobiological correlates of cue-induced craving in alcoholic patients and suggests that these correlates can be influenced by therapeutic interventions. The presence of emotional aspects of craving is suggested by amygdala activation. PMID- 11431230 TI - Five-year clinical course associated with DSM-IV alcohol abuse or dependence in a large group of men and women. AB - OBJECTIVE: The prognostic validity of the DSM-IV diagnoses of alcohol abuse and alcohol dependence was evaluated by examining the 5-year clinical course associated with those diagnoses in a large group of predominantly blue-collar men and women. METHOD: Personal semistructured interviews were carried out 5 years after an initial evaluation with 1,346 (75%) of the approximately 1,800 men and women participating in the Collaborative Study on the Genetics of Alcoholism who were eligible for follow-up. RESULTS: About two-thirds of the 298 subjects with DSM-IV alcohol dependence at baseline maintained that diagnosis during the 5-year study period. Fifty-five percent of the 288 subjects with DSM-IV alcohol abuse at baseline continued to meet one or more of the 11 DSM-IV abuse/dependence criteria, and 3.5% went on to meet the criteria for dependence at follow-up. Among the 760 subjects with no alcohol diagnosis at baseline, 2.5% met the criteria for alcohol dependence and 12.8% for alcohol abuse at follow-up. Baseline characteristics that predicted the occurrence of any of the 11 DSM-IV abuse/dependence criteria during the 5-year interval included male gender, lack of marital stability, presence of several of the criteria for dependence, and history of illicit drug use. CONCLUSIONS: The data suggest that over 5 years the DSM-IV diagnosis of alcohol dependence predicts a chronic disorder with a relatively severe course, while DSM-IV alcohol abuse predicts a less persistent, milder disorder that does not usually progress to dependence. PMID- 11431231 TI - A Norwegian psychiatric epidemiological study. AB - OBJECTIVE: This study reports results of a large-scale epidemiological investigation of the prevalence of mental disorder in Oslo. METHOD: A random sample of Oslo residents age 18-65 years was drawn from the Norwegian National Population Register. A total of 2,066 subjects, 57.5% of the original sample, were interviewed with the Composite International Diagnostic Interview in 1994 1997. The mean age of the interviewed subjects was 39.3 years. RESULTS: The 12 month prevalence of all mental disorders was 32.8%, and the lifetime prevalence was 52.4%. Alcohol abuse/dependence and major depression had the highest lifetime prevalence and 12-month prevalences. All mental disorders were more prevalent in women than in men, with the exception of alcohol and drug abuse/dependence. Severe psychopathology (e.g., three or more diagnoses) was found in 14%-15% of the respondents. The lifetime and 12-month prevalences for all diagnostic categories except drug abuse/dependence were similar to those found in the United States Comorbidity Survey. CONCLUSIONS: Epidemiological data for Oslo show that the lifetime and 12-month prevalences of mental disorder are quite high, with alcohol abuse/dependence and major depression particularly frequent. The rates for women are higher than those for men for all diagnostic categories, except for alcohol and drug abuse/dependence. PMID- 11431232 TI - Male criminals with organic brain syndrome: two distinct types based on age at first arrest. AB - OBJECTIVE: This study examined whether criminals with organic brain syndrome could be divided into two distinct types. The authors proposed that early starters (onset of criminal activity by age 18) would display a persistent, long lasting pattern of deviance that was largely independent of their brain disorder, whereas late starters (onset at age 19 or after) would exhibit deviant behaviors that began late in life and were more directly related to their brain disorder. METHOD: Subjects were 1,130 male criminal offenders drawn from a birth cohort of all individuals born between January 1, 1944, and December 31, 1947, in Denmark. The main study group included all men with both a history of criminal arrest and a hospitalization for organic brain syndrome (N=565). In addition, for a subset of analyses, the authors examined a randomly selected, same-size comparison group of men with a history of criminal arrest who were not hospitalized for organic brain syndrome. Data were available on all arrests and all psychiatric hospitalizations for individuals in this cohort through the age of 44. RESULTS: Among those with organic brain syndrome, early starters were significantly more likely than late starters to 1) be arrested before the onset of organic brain syndrome, 2) show a higher rate of offending before but not after the onset of organic brain syndrome, 3) be both recidivists and violent recidivists, and 4) have a diagnosis of antisocial personality disorder. CONCLUSIONS: Male criminals with organic brain syndrome can be meaningfully divided into two distinct types on the basis of age at first arrest. Early starters show a more global, persistent, and stable pattern of offending than late starters. These results have implications for treatment and risk assessment. PMID- 11431233 TI - Relation of prefrontal cortex dysfunction to working memory and symptoms in schizophrenia. AB - OBJECTIVE: The dorsolateral prefrontal cortex has been implicated in both working memory and the pathophysiology of schizophrenia. A relationship among dorsolateral prefrontal cortex activity, working memory dysfunction, and symptoms in schizophrenia has not been firmly established, partly because of generalized cognitive impairments in patients and task complexity. Using tasks that parametrically manipulated working memory load, the authors tested three hypotheses: 1) patients with schizophrenia differ in prefrontal activity only when behavioral performance differentiates them from healthy comparison subjects, 2) dorsolateral prefrontal cortex dysfunction is associated with poorer task performance, and 3) dorsolateral prefrontal cortex dysfunction is associated with cognitive disorganization but not negative or positive symptoms. METHOD: Seventeen conventionally medicated patients with schizophrenia and 16 healthy comparison subjects underwent functional magnetic resonance imaging while performing multiple levels of the "n-back" sequential-letter working memory task. RESULTS: Patients with schizophrenia showed a deficit in physiological activation of the right dorsolateral prefrontal cortex (Brodmann's area 46/9) in the context of normal task-dependent activity in other regions, but only under the condition that distinguished them from comparison subjects on task performance. Patients with greater dorsolateral prefrontal cortex dysfunction performed more poorly. Dorsolateral prefrontal cortex dysfunction was selectively associated with disorganization symptoms. CONCLUSIONS: These results are consistent with the hypotheses that working memory dysfunction in patients with schizophrenia is caused by a disturbance of the dorsolateral prefrontal cortex and that this disturbance is selectively associated with cognitive disorganization. Further, the pattern of behavioral performance suggests that dorsolateral prefrontal cortex dysfunction does not reflect a deficit in the maintenance of stimulus representations per se but points to deficits in more associative components of working memory. PMID- 11431234 TI - Effect of schizophrenia on frontotemporal activity during word encoding and recognition: a PET cerebral blood flow study. AB - OBJECTIVE: Neuropsychological studies have shown that deficits in verbal episodic memory in schizophrenia occur primarily during encoding and retrieval stages of information processing. The current study used positron emission tomography to examine the effect of schizophrenia on change in cerebral blood flow (CBF) during these memory stages. METHOD: CBF was measured in 23 healthy comparison subjects and 23 patients with schizophrenia during four conditions: resting baseline, motor baseline, word encoding, and word recognition. The motor baseline was used as a reference that was subtracted from encoding and recognition conditions by using statistical parametric mapping. RESULTS: Patients' performance was similar to that of healthy comparison subjects. During word encoding, patients showed reduced activation of left prefrontal and superior temporal regions. Reduced left prefrontal activation in patients was also seen during word recognition, and additional differences were found in the left anterior cingulate, left mesial temporal lobe, and right thalamus. Although patients' performance was similar to that of healthy comparison subjects, left inferior prefrontal activation was associated with better performance only in the comparison subjects. CONCLUSIONS: Left frontotemporal activation during episodic encoding and retrieval, which is associated with better recognition in healthy people, is disrupted in schizophrenia despite relatively intact recognition performance and right prefrontal function. This may reflect impaired strategic use of semantic information to organize encoding and facilitate retrieval. PMID- 11431235 TI - Dysfunction of early-stage visual processing in schizophrenia. AB - OBJECTIVE: Schizophrenia is associated with deficits in higher-order processing of visual information. This study evaluated the integrity of early visual processing in order to evaluate the overall pattern of visual dysfunction in schizophrenia. METHOD: Steady-state visual-evoked potential responses were recorded over the occipital cortex in patients with schizophrenia and in age- and sex-matched comparison volunteers. Visual-evoked potentials were obtained for stimuli composed of isolated squares that were modulated sinusoidally in luminance contrast, number of squares, or chromatic contrast in order to emphasize magnocellular or parvocellular visual pathway activity. RESULTS: Responses of patients to magnocellular-biased stimuli were significantly lower than those of comparison volunteers. These lower response levels were observed in conditions using both low luminance contrast and large squares that biased processing toward the magnocellular pathway. In contrast, responses to stimuli that biased processing toward the parvocellular pathway were not significantly different between schizophrenia patients and comparison volunteers. A significant interaction of group and stimulus type was observed in the condition using low luminance contrast. CONCLUSIONS: These findings suggest a dysfunction of lower level visual pathways, which was more prominent for magnocellular than parvocellular biased stimuli. The magnocellular pathway helps in orienting toward salient stimuli. A magnocellular pathway deficit could contribute to higher-level visual cognitive deficits in schizophrenia. PMID- 11431236 TI - Association of estrogen levels with neuropsychological performance in women with schizophrenia. AB - OBJECTIVE: This study sought to determine the relationship of estrogen levels with psychiatric symptoms and neuropsychological function in female patients with schizophrenia. METHOD: Psychiatric symptoms were assessed and average estrogen and progesterone levels from four consecutive weekly blood samples were measured in 22 female inpatients with schizophrenia who were also administered a neuropsychological battery. RESULTS: There were strong positive correlations between average estrogen level and cognitive function, especially measures of global cognitive function, verbal and spatial declarative memory, and perceptual motor speed. Correlations of hormone levels with psychiatric symptoms were nonsignificant. CONCLUSIONS: Higher estrogen levels in female patients with schizophrenia are associated with better cognitive ability. These results may have implications for potential treatment of cognitive dysfunction with adjunctive estrogen in female patients with schizophrenia. PMID- 11431237 TI - Structural brain abnormalities in chronic schizophrenia at the extremes of the outcome spectrum. AB - OBJECTIVE: This study investigated the relationship between outcome and structural brain abnormalities in schizophrenia. METHOD: Intracranial volume and volumes of the cerebrum, gray and white matter, lateral and third ventricles, frontal lobes, thalamus, and cerebellum were measured in 20 patients with a poor outcome, 25 with a favorable outcome, and 23 healthy comparison subjects with magnetic resonance imaging. RESULTS: Thalamic volume was significantly smaller both in poor-outcome patients and good-outcome patients. In contrast, only poor outcome patients displayed significantly smaller cerebral gray matter, particularly prefrontal, and enlargement of the lateral and third ventricles. No significant differences were found for intracranial, cerebellar, or cortical CSF volumes. CONCLUSIONS: Smaller thalamic volumes in schizophrenia may reflect a greater susceptibility for the disorder and seem unrelated to outcome. In contrast, gray matter volume loss of the cerebrum, particularly in the frontal lobes, and lateral and third ventricular enlargement appear related to outcome in schizophrenia. PMID- 11431238 TI - Right prefrontal repetitive transcranial magnetic stimulation in obsessive compulsive disorder: a double-blind, placebo-controlled study. AB - OBJECTIVE: The efficacy of repetitive transcranial magnetic stimulation (rTMS) of the right prefrontal cortex for patients with obsessive-compulsive disorder (OCD) was studied under double-blind, placebo-controlled conditions. METHOD: Patients were randomly assigned to 18 sessions of real (N=10) or sham (N=8) rTMS. Treatments lasted 20 minutes, and the frequency was 1 Hz for both conditions, but the intensity was 110% of motor threshold for real rTMS and 20% for the sham condition. RESULTS: No significant changes in OCD were detected in either group after treatment. Two patients who received real rTMS, with checking compulsions, and one receiving sham treatment, with sexual/religious obsessions, were considered responders. CONCLUSIONS: Low-frequency rTMS of the right prefrontal cortex failed to produce significant improvement of OCD and was not significantly different from sham treatment. Further studies are indicated to assess the efficacy of rTMS in OCD and to clarify the optimal stimulation characteristics. PMID- 11431239 TI - Treatment of panic attack and risk of major depressive disorder in the community. AB - OBJECTIVE: The authors' goal was to determine whether treatment of panic attacks has a protective effect on the risk of major depression in the community. METHOD: Data were drawn from the National Comorbidity Survey, a community-based household sample representative of the U.S. adult population. A Cox proportional hazard model was used to estimate the association between risk of first-onset major depression and panic among subjects who had or had not received treatment for panic. RESULTS: A significantly smaller proportion of individuals who received treatment for panic (19%) than those who did not receive treatment (45%) developed major depression. This difference remained significant in a Cox proportional hazard analysis adjusted for age at onset of panic and differences in demographic characteristics. CONCLUSIONS: Detection and treatment of panic may reduce the risk of developing major depression. PMID- 11431240 TI - Double-blind, placebo-controlled comparison of intramuscular olanzapine and intramuscular haloperidol in the treatment of acute agitation in schizophrenia. AB - OBJECTIVE: The authors evaluated the comparative efficacy and safety of intramuscular olanzapine, intramuscular haloperidol, and intramuscular placebo for the treatment of acute agitation in schizophrenia. METHOD: Hospitalized patients with schizophrenia received one to three injections of intramuscular olanzapine, 10 mg, intramuscular haloperidol, 7.5 mg, or intramuscular placebo over a 24-hour period. Agitation was measured with the excited component of the Positive and Negative Syndrome Scale and two additional scales. RESULTS: According to scores on the excited component of the Positive and Negative Syndrome Scale, both intramuscular olanzapine and intramuscular haloperidol reduced agitation significantly more than intramuscular placebo 2 and 24 hours following the first injection. Intramuscular olanzapine reduced agitation significantly more than intramuscular haloperidol 15, 30, and 45 minutes following the first injection. No patients treated with intramuscular olanzapine experienced acute dystonia, compared with 7% of those who were treated with intramuscular haloperidol. No significant QT(c) interval changes were observed in any patients. CONCLUSIONS: Intramuscular olanzapine represents a rapid, effective, and safe treatment for acute agitation in schizophrenia. PMID- 11431241 TI - Altered serotonin 2A receptor activity in women who have recovered from bulimia nervosa. AB - OBJECTIVE: The authors' goal was to confirm that brain serotonin (5-HT) alterations are present in patients who have recovered from bulimia nervosa. Positron emission tomography imaging with [(18)F]altanserin was used to characterize binding of the 5-HT(2A) receptor, which might contribute to altered feeding, mood, or impulse control. METHOD: Nine women who had recovered from bulimia nervosa (they had no episodes of binge eating or purging, were at normal weight, and had regular menstrual cycles for more than 1 year) were compared with 12 female volunteers who had never had bulimia. RESULTS: The healthy volunteers, but not the women who had recovered from bulimia nervosa, had an age-related decline in 5-HT(2A) binding. Women who had recovered from bulimia nervosa had a reduction of medial orbital frontal cortex 5-HT(2A) binding. CONCLUSIONS: The lack of age-related changes in 5-HT activity is further evidence of 5-HT alterations in subjects who have recovered from bulimia nervosa. In addition, vulnerabilities for eating disorders, impulse dyscontrol, and mood disturbances may involve 5-HT and frontal lobe activity. PMID- 11431242 TI - A comparison of family functioning, temperament, and childhood conditions in monozygotic twin pairs discordant for lifetime bulimia nervosa. AB - OBJECTIVE: The authors investigated differences between twins in nine pairs of female monozygotic twins in the Australian Twin Registry who were discordant for lifetime bulimia nervosa. METHOD: The twins affected and unaffected by lifetime bulimia nervosa were compared on self-report measures, including a measure of parental bonding, four measures of temperament, and six early-childhood medical conditions. RESULTS: No twins had current bulimia nervosa, and there was no difference in weight or eating status between the affected and unaffected twins. The affected twins reported significantly lower self-esteem and less warmth but more overprotection by their mothers during childhood. CONCLUSIONS: Although limited by the small number of discordant twin pairs and the inability to detect causal relationships, these results suggest that environmental influences that promote low self-esteem may also increase the risk for bulimia nervosa. These temperamental differences may explain the discrepancies in parenting or perceived parenting. PMID- 11431243 TI - Charles Bonnet syndrome and multiple sclerosis. PMID- 11431244 TI - Olanzapine-induced somnambulism. PMID- 11431245 TI - Venlafaxine-induced hair loss. PMID- 11431246 TI - Prodissociative effects of metyrapone. PMID- 11431247 TI - Virtual psychiatric clinics. PMID- 11431248 TI - E-maternity leave. PMID- 11431249 TI - Untreated initial psychosis. PMID- 11431250 TI - Childhood chronic fatigue syndrome. PMID- 11431252 TI - Cytokines in clinical psychiatry. PMID- 11431254 TI - Comparing psychotherapy and pharmacotherapy. PMID- 11431255 TI - Comparing psychotherapy and pharmacotherapy. PMID- 11431256 TI - Comparing psychotherapy and pharmacotherapy. PMID- 11431259 TI - Reformulating the diagnosis of schizophrenia. PMID- 11431260 TI - Respect from our medical colleagues. PMID- 11431261 TI - Respect from our medical colleagues. PMID- 11431263 TI - Growth of interest in personality disorders. PMID- 11431265 TI - DNA polymorphisms and bipolar disorder. PMID- 11431266 TI - Unusual book review. PMID- 11431280 TI - Is the two week rule for cancer referrals working? PMID- 11431281 TI - Adoption. PMID- 11431282 TI - The continuing global challenge of injury. PMID- 11431283 TI - Iatrogenic vCJD from surgical instruments. PMID- 11431284 TI - Fatigue in cancer. PMID- 11431285 TI - UK attacked for lack of injury prevention strategy. PMID- 11431286 TI - AMA split by dispute between chief and board of trustees. PMID- 11431288 TI - Guidance issued on feeding of sick newborn babies. PMID- 11431289 TI - Flexible jobs needed to retain women doctors. PMID- 11431290 TI - Minorities get preferential admission to US medical schools. PMID- 11431291 TI - Report attacked for vested interests of its backers. PMID- 11431292 TI - Dutch patients travel to Spain for orthopaedic surgery. PMID- 11431293 TI - Public inquiry hears how Shipman killed patients with diamorphine. PMID- 11431294 TI - Ban on human reproductive cloning demanded. PMID- 11431296 TI - Effect of zinc supplementation on malaria and other causes of morbidity in west African children: randomised double blind placebo controlled trial. AB - OBJECTIVE: To study the effects of zinc supplementation on malaria and other causes of morbidity in young children living in an area holoendemic for malaria in west Africa. DESIGN: Randomised, double blind, placebo controlled efficacy trial. SETTING: 18 villages in rural northwestern Burkina Faso. PARTICIPANTS: 709 children were enrolled; 685 completed the trial. INTERVENTION: Supplementation with zinc (12.5 mg zinc sulphate) or placebo daily for six days a week for six months. MAIN OUTCOME MEASURES: The primary outcome was the incidence of symptomatic falciparum malaria. Secondary outcomes were the severity of malaria episodes, prevalence of malaria parasite, mean parasite densities, mean packed cell volume, prevalence of other morbidity, and all cause mortality. RESULTS: The mean number of malaria episodes per child (defined as a temperature >/=37.5 degrees C with >/=5000 parasites/microliter) was 1.7, 99.7% due to infection with Plasmodium falciparum. No difference was found between the zinc and placebo groups in the incidence of falciparum malaria (relative risk 0.98, 95% confidence interval 0.86 to 1.11), mean temperature, and mean parasite densities during malaria episodes, nor in malaria parasite rates, mean parasite densities, and mean packed cell volume during cross sectional surveys. Zinc supplementation was significantly associated with a reduced prevalence of diarrhoea (0.87, 0.79 to 0.95). All cause mortality was non-significantly lower in children given zinc compared with those given placebo (5 v 12, P=0.1). CONCLUSIONS: Zinc supplementation has no effect on morbidity from falciparum malaria in children in rural west Africa, but it does reduce morbidity associated with diarrhoea. PMID- 11431298 TI - Randomised trial of cranberry-lingonberry juice and Lactobacillus GG drink for the prevention of urinary tract infections in women. AB - OBJECTIVE: To determine whether recurrences of urinary tract infection can be prevented with cranberry-lingonberry juice or with Lactobacillus GG drink. DESIGN: Open, randomised controlled 12 month follow up trial. SETTING: Health centres for university students and staff of university hospital. PARTICIPANTS: 150 women with urinary tract infection caused by Escherichia coli randomly allocated into three groups. INTERVENTIONS: 50 ml of cranberry-lingonberry juice concentrate daily for six months or 100 ml of lactobacillus drink five days a week for one year, or no intervention. MAIN OUTCOME MEASURE: First recurrence of symptomatic urinary tract infection, defined as bacterial growth >/=10(5 )colony forming units/ml in a clean voided midstream urine specimen. RESULTS: The cumulative rate of first recurrence of urinary tract infection during the 12 month follow up differed significantly between the groups (P=0.048). At six months, eight (16%) women in the cranberry group, 19 (39%) in the lactobacillus group, and 18 (36%) in the control group had had at least one recurrence. This is a 20% reduction in absolute risk in the cranberry group compared with the control group (95% confidence interval 3% to 36%, P=0.023, number needed to treat=5, 95% confidence interval 3 to 34). CONCLUSION: Regular drinking of cranberry juice but not lactobacillus seems to reduce the recurrence of urinary tract infection. PMID- 11431299 TI - Randomised trial of acupuncture compared with conventional massage and "sham" laser acupuncture for treatment of chronic neck pain. AB - OBJECTIVES: To compare the efficacy of acupuncture and conventional massage for the treatment of chronic neck pain. DESIGN: Prospective, randomised, placebo controlled trial. SETTING: Three outpatient departments in Germany. PARTICIPANTS: 177 patients aged 18-85 years with chronic neck pain. INTERVENTIONS: Patients were randomly allocated to five treatments over three weeks with acupuncture (56), massage (60), or "sham" laser acupuncture (61). MAIN OUTCOME MEASURES: PRIMARY OUTCOME MEASURE: maximum pain related to motion (visual analogue scale) irrespective of direction of movement one week after treatment. SECONDARY OUTCOME MEASURES: range of motion (3D ultrasound real time motion analyser), pain related to movement in six directions (visual analogue scale), pressure pain threshold (pressure algometer), changes of spontaneous pain, motion related pain, global complaints (seven point scale), and quality of life (SF-36). Assessments were performed before, during, and one week and three months after treatment. Patients' beliefs in treatment were assessed. RESULTS: One week after five treatments the acupuncture group showed a significantly greater improvement in motion related pain compared with massage (difference 24.22 (95% confidence interval 16.5 to 31.9), P=0.0052) but not compared with sham laser (17.28 (10.0 to 24.6), P=0.327). Differences between acupuncture and massage or sham laser were greater in the subgroup who had had pain for longer than five years (n=75) and in patients with myofascial pain syndrome (n=129). The acupuncture group had the best results in most secondary outcome measures. There were no differences in patients' beliefs in treatment. CONCLUSIONS: Acupuncture is an effective short term treatment for patients with chronic neck pain, but there is only limited evidence for long term effects after five treatments. PMID- 11431300 TI - Specialist registrars' plans for working part time as consultants in medical specialties: questionnaire study. PMID- 11431301 TI - Primary care groups: Improving the quality of care through clinical governance. PMID- 11431302 TI - Regular review: Managing testicular cancer. PMID- 11431303 TI - Lesson of the week: Depot corticosteroid treatment for hay fever causing avascular necrosis of both hips. PMID- 11431304 TI - Lesson of the week: Reye's syndrome and aspirin: lest we forget. PMID- 11431305 TI - For and against: Public health does not need to be led by doctors. For. PMID- 11431306 TI - Summarising economic evaluations in systematic reviews: a new approach. PMID- 11431319 TI - Extraction of 5-fluorouracil by tumor and liver: a noninvasive positron emission tomography study of patients with gastrointestinal cancer. AB - Tumor and normal tissue pharmacokinetics of 5-Fluorouracil (5-FU) in patients can be determined with positron emission tomography scanning. However, the data obtained are of limited value because of the inability to distinguish catabolites (inactive species) from parent 5-FU and anabolites (cytotoxic species). In this paper, we have blocked 5-FU catabolism in one arm of a paired study with eniluracil, an inactivator of dihydropyrimidine dehydrogenase, enabling catabolite correction and calculation of tissue pharmacokinetic parameters to be achieved. Using this novel approach, we report for the first time that the net clearance of 5-[(18)F]FU from plasma into tumors (liver metastases and pancreatic tumor) of patients is low (K(I) = 0.0033 +/- 0.0005 ml plasma/ml tissue/min). In contrast, the initial (up to 10 min) clearance through catabolism in liver was high (K(I) = 0.7313 +/- 0.092 ml plasma/ml tissue/min). In the absence of eniluracil, catabolites in tumors accounted for 83% of total tumor exposure (range, 66-91%), whereas catabolites in liver accounted for 96% of total liver exposure (range, 94-98%). This study provides definitive evidence that the cytotoxicity of 5-FU in patients with gastrointestinal cancer could be compromised by its intrinsically low uptake by tumors, as well as decreased systemic availability through hepatic catabolism. PMID- 11431320 TI - Mutations of tumor necrosis factor-related apoptosis-inducing ligand receptor 1 (TRAIL-R1) and receptor 2 (TRAIL-R2) genes in metastatic breast cancers. AB - Several lines of evidence suggest that apoptosis dysregulation plays an important role in cancer metastasis. In this study, to explore the possibility that the mutations of death receptors are involved in the metastasis mechanism, we analyzed the death domains of Fas and tumor necrosis factor-related apoptosis inducing ligand-receptor 1 and -2 (TRAIL-R1 and -R2) genes for the detection of somatic mutations in 57 breast cancers with (n = 34) or without (n = 23) metastasis to the regional lymph nodes. We found seven mutations (three TRAIL-R1 and four TRAIL-R2 mutations), and these mutations were detected only in the breast cancers with metastasis. Furthermore, we also analyzed the allelic losses of chromosome 8p21-22, where TRAIL-R1 and R2 reside in the same series of breast cancers, and found that the allelic losses were significantly higher in metastatic breast cancers. We expressed the tumor-derived TRAIL-R1 and TRAIL-R2 mutants in 293 cells and found that apoptosis was suppressed. These data suggest that TRAIL-R1 and R2 genes are relevant to the frequent loss of chromosome 8p21 22 in breast cancer and that the inactivating mutations of TRAIL-R1 and -R2 genes play a role in the metastasis of breast cancer. PMID- 11431321 TI - Loss of imprinting of insulin-like growth factor-II in Wilms' tumor commonly involves altered methylation but not mutations of CTCF or its binding site. AB - Loss of imprinting (LOI) is the most common molecular abnormality in Wilms' tumor (WT), other embryonal cancers, and most other tumor types. LOI in WT involves activation of the normally silent maternal allele of the insulin-like growth factor-II (IGF2) gene, silencing of the normally active maternal allele of the H19 gene, and aberrant methylation of a differentially methylated region (DMR) upstream of the maternal copy of H19. Recently, the transcription factor CTCF, which binds to the H19 DMR, has been implicated in the maintenance of H19 and IGF2 imprinting. Here, we show that mutations in the CTCF gene or in the H19 DMR do not occur at significant frequency in WT, nor is there transcriptional silencing of CTCF. We also confirm that methylation of the H19 DMR in WT with LOI includes the CTCF core consensus site. However, some WTs with normal imprinting of IGF2 also show aberrant methylation of CTCF binding sites, indicating that methylation of these sites is necessary but not sufficient for LOI in WT. PMID- 11431322 TI - Structural analysis of the 17q22-23 amplicon identifies several independent targets of amplification in breast cancer cell lines and tumors. AB - A novel region of amplification in breast tumors has recently been identified on chromosome 17q22-23. In an effort to identify the oncogenes in the region that are targeted by the amplification process, we determined the structure of the amplicon in breast cancer cell lines and tumors. Physical and transcription maps of the approximately 3.5-Mb region were established and used as the basis for copy number analysis within the region by Southern blot and fluorescence in situ hybridization. Seven specific and independent amplification maxima were identified in breast cancer cell lines and breast tumors. We present correlative amplification and overexpression studies for the FLJ21316 and Hs.6649 genes suggesting a role for these candidates as amplification-dependent oncogenes. PMID- 11431323 TI - Formation of intracranial tumors by genetically modified human astrocytes defines four pathways critical in the development of human anaplastic astrocytoma. AB - The formation of human malignant gliomas is thought to involve the accumulation of multiple genetic alterations. To define the function of specific alterations in glioma formation, we serially introduced genetic alterations functionally equivalent to those noted in human malignant gliomas into normal human astrocytes (NHAs). We then monitored the ability of each of these alterations to contribute to the growth of otherwise genetically stable NHAs into intracranial malignant gliomas. Using this model, we show that expression of human telomerase catalytic component (hTERT), but not E7-mediated inactivation of pRb or E6/E7-mediated inactivation of p53/pRb, was sufficient to initiate the tumorigenic process by circumventing cellular senescence in astrocytes. hTERT expression, even in combination with inactivation of p53/pRb, did not transform astrocytes. These alterations together, however, cooperated with ras pathway activation (initiated by expression of mutant H-Ras), but not with phosphatidylinositol 3-kinase pathway activation (initiated by expression of myristoylated Akt) or epidermal growth factor receptor activation, to allow for the formation of intracranial tumors strongly resembling p53/pRb pathway-deficient, telomerase-positive, ras activated human grade III anaplastic astrocytomas. These results identify four pathways as key in the development of human anaplastic astrocytomas. PMID- 11431324 TI - Epithelial cancer cell migration: a role for chemokine receptors? AB - We investigated the possibility that chemokine gradients influence migration of human ovarian epithelial tumor cells. Of 14 chemokine receptors investigated, only CXCR4 was expressed on ovarian cancer cells. CXCR4 mRNA localized to a subpopulation of tumor cells in ovarian cancer biopsies. Ovarian cancer cell lines and cells freshly isolated from ascites expressed CXCR4 protein. The CXCR4 ligand, CXCL12, was found in ascites from 63 patients. CXCL12 elicited intracellular calcium flux and directed migration and changes in integrin expression in ovarian cancer cells. CXCR4 may influence cell migration in the peritoneum, a major route for ovarian cancer spread, and could be a therapeutic target. PMID- 11431325 TI - Dynamic micro-magnetic resonance imaging of liver micrometastasis in mice with a novel liver macromolecular magnetic resonance contrast agent DAB-Am64-(1B4M Gd)(64). AB - DAB-Am64-(1B4M-Gd)(64) is a newly synthesized macromolecular liver magnetic resonance imaging (MRI) contrast agent with a polypropylenimine diaminobutane (DAB) dendrimer conjugated with a bifunctional diethylenetriaminepentaacetic acid (DTPA) derivative for complexing Gd(III) atoms. The characteristics of DAB-Am64 (1B4M-Gd)(64), which quickly accumulated in the liver, have been reported recently. In the present study, the dynamic micro-MRI with DAB-Am64-(1B4M-Gd)(64) was obtained in the mouse liver metastasis model using colon carcinoma cells to evaluate the ability to visualize the micrometastatic tumors compared with that using Gd-DTPA. The dynamic micro-MRI with DAB-Am64-(1B4M-Gd)(64) was able to homogeneously enhance the normal liver parenchyma and visualize micrometastatic tumors of 0.3-mm diameter in the liver of the mice with better contrast than that with Gd-DTPA. In conclusion, DAB-Am64-(1B4M-Gd)(64) is a new liver MRI contrast agent potentially useful for diagnosis of micrometastasis in the liver. PMID- 11431326 TI - Monitoring response to convection-enhanced taxol delivery in brain tumor patients using diffusion-weighted magnetic resonance imaging. AB - Convection-enhanced drug delivery (CEDD) is a novel approach to enhance the delivery of drugs directly into brain tumors. We have used diffusion-weighted MRI (DWMRI) to monitor the effects of intratumoral CEDD in three brain tumor patients treated with Taxol. Clear changes in the images and the water diffusion parameters were observed shortly after the initiation of treatment. Initially, a bright area corresponding to decreased diffusion appeared, followed by the appearance of a dark area of increased diffusion within the bright area. The time to appearance of the dark area varied among the patients, suggesting different response rates. In this work, we have demonstrated the feasibility of using DWMRI as a noninvasive tool to achieve unique early tissue characterization not attainable by other conventional imaging methods. PMID- 11431327 TI - The tumor suppressor candidate p33(ING1) mediates repair of UV-damaged DNA. AB - The biological functions of the tumor suppressor, ING1, have been studied extensively in the last 5 years since it was cloned. It shares many biological functions with those of p53 and has been reported to mediate growth arrest, senescence, apoptosis, anchorage-dependent growth, and chemosensitivity. Some of these functions, such as cell cycle arrest and apoptosis, have been shown to be dependent on the activity of both ING1 and p53 proteins. In this study, we report that p33(ING1) (one of ING1 isoforms) is also involved in the modulation of DNA repair. We found that overexpression of p33(ING1) enhances repair of UV-damaged DNA and that p53 is required for the repair process. Furthermore, binding between ING1 and GADD45 has been detected. These observations suggest that p33(ING1) cooperates with p53 in nucleotide excision repair and that GADD45 may be one of its components. PMID- 11431328 TI - Loss of focal adhesion kinase (FAK) inhibits epidermal growth factor receptor dependent migration and induces aggregation of nh(2)-terminal FAK in the nuclei of apoptotic glioblastoma cells. AB - In glioblastoma cells, inhibition of focal adhesion kinase (FAK) by the focal adhesion targeting domain attenuated epidermal growth factor receptor (EGFR) signaling, inhibiting epidermal growth factor-dependent migration. Although the EGFR-specific antagonist PD153035 increased caspase-3 activity, this was independent of FAK activity. Instead, the increase in apoptosis upon inhibition of FAK induced the aggregation of an NH(2)-terminal FAK fragment normally present in the nucleus. A recombinant NH(2)-terminal FAK construct was also targeted to the nucleus and aggregated in apoptotic cells upon coexpression with the focal adhesion targeting domain. Therefore, loss of FAK from the focal adhesions inhibits EGFR signaling at the cell membrane and transmits a proapoptotic signal to an NH(2)-terminal variant of FAK present in the nucleus. PMID- 11431329 TI - Polymorphisms in the prostate cancer susceptibility gene HPC2/ELAC2 in multiplex families and healthy controls. AB - Two polymorphisms in the newly cloned prostate cancer susceptibility gene, HPC2/ELAC2, are suspected to be associated with an increased risk of developing the disease. These missense variants result in a serine (S) to leucine (L) substitution at amino acid residue 217 and an alanine (A) to threonine (T) substitution at residue 541. We genotyped these polymorphisms in 257 multiplex prostate cancer sibships and in 355 race-matched healthy unrelated controls. A significant increase in the frequency of the T allele is seen in the prostate cancer subjects compared with controls. There is, however, little evidence for excess clustering of the T allele within the multiplex families known to be segregating this allele, and there is no evidence for linkage of prostate cancer to the HPC2/ELAC2 region of chromosome 17p11.2 in these families. The T allele shows no association with either Gleason score or age-of-onset in segregating families. PMID- 11431330 TI - Regulation of PTEN binding to MAGI-2 by two putative phosphorylation sites at threonine 382 and 383. AB - We have reported previously that the PTEN COOH-terminal 33 amino acids play a role in the maintenance of PTEN protein stability (Tolkacheva and Chan, Oncogene, 19: 680-689, 2000). By site-directed mutagenesis, we identified two threonine residues within this COOH-terminal region at codon 382 and 383 that may be targets for phosphorylation events. Interestingly, PTEN mutants rendered phosphorylation-incompetent at these two sites, T382A/T383A, and were found to have drastically reduced expression in cultured cells. The enhanced degradation of PTEN was most likely mediated by the proteosome-dependent pathway, we have evidence that PTEN was polyubiquitinated. More interestingly, the non phosphorylated forms of PTEN displayed significantly greater binding affinity than the wild-type protein to a previously identified PTEN interacting partner, MAGI-2/ARIP1. On the basis of all these data, we propose that PTEN recruitment to the cell-cell junction may be regulated through the phosphorylation of its COOH terminus. PMID- 11431331 TI - DNA damage-activated kinase Chk2 is independent of proliferation or differentiation yet correlates with tissue biology. AB - The Chk2 kinase is a tumor suppressor and key transducer of DNA-damage checkpoints. We show that the human Chk2 protein is relatively stable, nuclear, and responding to gamma-radiation throughout the cell cycle. Contrary to the retinoblastoma protein-regulated, labile Chk1 kinase restricted to S-G(2) phases, Chk2 remains activatable even in quiescent and differentiating cells. In human tissues, Chk2 is homogeneously expressed in renewing cell populations such as epidermis or intestine, heterogeneous in conditionally renewing tissues, and absent or cytoplasmic in static tissues such as muscle or brain. These data highlight striking differences between Chk2 and Chk1 and show unexpected correlation of Chk2 expression with tissue biology. PMID- 11431332 TI - Runt-related gene 2 in endothelial cells: inducible expression and specific regulation of cell migration and invasion. AB - Understanding the regulation of endothelial cell (EC) gene expression has important implications for angiogenesis, tumor growth, and metastasis. The transcription factor runt-related gene 2 (RUNX2)/core binding factoralpha-1/acute myeloid leukemia 3/polyoma enhancer-binding protein 2alphaA/osteoblast-specific transcription factor 2 regulates osteoblast differentiation, increases lymphomagenesis in transgenic mice, and is expressed in murine ECs. Here, we report on RUNX2 expression in human bone marrow EC (HBME-1) and its role in EC differentiation. Expression of RUNX2 occurred in HBME-1 cultured on extracellular matrix (ECM) substrates that stimulate in vitro differentiation (tube formation). Neutralizing anti-insulin-like growth factor (IGF)-I-receptor antibody inhibited tube formation as well as activation of RUNX2 expression in HBME-1 cultured on ECM. IGF-I treatment also increased both RUNX2 mRNA and protein expression. HBME 1 transfectants expressing dominant-negative (DN) RUNX were established to address the role of RUNX2 in these processes. HBME/DN cells exhibited reduced tube formation activity relative to control transfectants and less ability to growth arrest and differentiate on ECM. DNRUNX expression also inhibited HBME-1 migration and invasion, which are necessary for tube formation. The urokinase type plasminogen activator and membrane-type MMP-1 genes were consistently down regulated in DNRUNX transfectants. The results suggest that RUNX2 is important in IGF-I and ECM-regulated EC migration and differentiation. RUNX2 effects on HBME-1 migration and invasion may occur through activation of protease expression, events that regulate angiogenesis, and tumor growth. PMID- 11431333 TI - Inhibitory effects of orally administered green tea, black tea, and caffeine on skin carcinogenesis in mice previously treated with ultraviolet B light (high risk mice): relationship to decreased tissue fat. AB - Treatment of SKH-1 hairless mice with ultraviolet B light (UVB; 30 mJ/cm(2)) twice a week for 22 weeks resulted in tumor-free animals with a high risk of developing malignant and nonmalignant skin tumors during the next several months in the absence of additional UVB treatment (high-risk mice). Oral administration of green tea or black tea (6 mg tea solids/ml) to UVB-pretreated high-risk SKH-1 mice for 23 weeks after stopping UVB treatment decreased the number of tumors/mouse, decreased the size of the parametrial fat pads, and decreased the thickness of the dermal fat layer away from tumors and directly under tumors. Administration of the decaffeinated teas had little or no effect on these parameters, and adding caffeine (equivalent to the amount in the regular teas) to the decaffeinated teas restored their inhibitory effects. Administration of caffeine alone also decreased the number of tumors/mouse, the size of the parametrial fat pads, and the thickness of the dermal fat layer away from tumors and under tumors. Using data from individual mice and linear regression and correlation analysis, we found a highly significant positive correlation between the thickness of the dermal fat layer away from tumors and the number of tumors/mouse (r = 0.34; P = 0.0001), but the correlation between average tumor size/mouse and the thickness of the dermal fat layer away from tumors was weak (r = 0.16; P = 0.034). The results suggested that p.o. administered tea or caffeine may have decreased tumor multiplicity in part by decreasing fat levels in the dermis. Additional analysis revealed that oral administration of caffeinated beverages (green tea, black tea, decaffeinated green tea plus caffeine, decaffeinated black tea plus caffeine, or caffeine alone) decreased the thickness of the dermal fat layer under large tumors to a much greater extent than under small tumors. This is the first demonstration of a close association between inhibition of carcinogenesis and the lowering of tissue fat levels by a chemopreventive agent. PMID- 11431334 TI - Qualitative and quantitative relationship between dysplastic aberrant crypt foci and tumorigenesis in the Min/+ mouse colon. AB - The multiple intestinal neoplasia (Min)/+ mouse, which harbors only one functional allele of the Apc gene, is susceptible to environmental factors that disrupt this gene and subsequently trigger Apc-driven tumorigenesis in the colon. Aberrant crypt foci (ACF) are assumed to be preneoplastic lesions in colon carcinogenesis. Recently, we reported the absence of "classical" ACF in the colon of untreated Min/+ mice. Instead we identified flat dysplastic lesions, which we denoted ACF(Min) (J. E. Paulsen et al., Scand. J. Gastroenterol., 35: 534-539, 2000). In contrast to the classical type, ACF(Min) are not elevated above the surrounding mucosa, and their detection is totally dependent on methylene blue staining and transillumination. In the present study, we treated Min/+ mice with 5 mg/kg body weight azoxymethane (AOM) at weeks 1 and 2 and demonstrated induction of both types of lesions. However, only ACF(Min) appeared to be associated with the development of adenomas. Monocryptal ACF(Min), large ACF(Min), and adenomas showed a uniform histopathological picture of dysplasia and cytoplasmic overexpression of beta-catenin, indicating a qualitative relationship between these lesions. Also a quantitative relationship was suggested because the dramatic decrease in ACF(Min) number from week 7 to 11 was paralleled by a reciprocal increase in tumor number, indicating fast-crypt multiplication of ACF(Min). In AOM-treated +/+ (wild-type) littermates, a low number of ACF(Min) and tumors with the same characteristics as in Min/+ mice was seen. In contrast to ACF(Min), histopathological and immunohistochemical examination of classical ACF showed normal or hyperplastic crypts with normal levels of beta-catenin expression. In AOM-treated Min/+ mice, the number of classical ACF was virtually constant from week 7 to 11, and only a modest increase of crypt multiplicity was observed. The number of AOM-induced classical ACF at week 11 was not different in Min/+ mice and +/+ mice. In conclusion, we identified two distinct populations of altered crypts in the colon of Min/+ mice after AOM treatment. The ACF(Min), which resemble the dysplastic ACF described previously, clearly showed a continuous development from the monocryptal stage to adenoma, and they were characterized by fast-growing crypts with altered control of beta-catenin. In contrast, the classical ACF, which resemble the hyperplastic ACF described previously, were characterized by slow-growing crypts with normal beta-catenin expression, and they were probably not related to tumorigenesis. PMID- 11431335 TI - Hepatic hyperplasia in noncirrhotic fatty livers: is obesity-related hepatic steatosis a premalignant condition? AB - It is not known whether obesity increases the risk for hepatocellular carcinoma (HCC) simply because it promotes cirrhosis, a general risk factor for HCC, or via some other mechanism that operates independently of cirrhosis. If the latter occurs, then hepatocyte hyperplasia, an early event during the neoplastic process, might begin before liver cirrhosis develops. Genetically obese, leptin deficient ob/ob mice are models for nonalcoholic fatty liver disease (NAFLD), a type of liver disease that is strongly associated with obesity and type 2 diabetes. Similar to obese, diabetic patients, ob/ob mice have an increased incidence of HCC. However, unlike humans with NAFLD, they rarely, if ever, develop cirrhosis spontaneously. To determine whether the noncirrhotic livers of ob/ob mice with NAFLD exhibit hepatocyte hyperplasia, parameters of proliferation and apoptosis were compared in adult ob/ob mice and their healthy litter mates. Adult ob/ob mice have an increase in liver mass relative to body mass. This hepatomegaly cannot be explained solely by lipid accumulation and is accompanied by significant increases in hepatocyte proliferative activity (as evidenced by increased Erk activation, cell-cycle related gene expression, bromodeoxyuridine incorporation, and hepatic DNA content) with concomitant inhibition of hepatocyte apoptosis (as evidenced by decreased numbers of apoptotic hepatocytes, induction of several antiapoptotic mechanisms, and decreased activation of procaspase 3). Thus, liver hyperplasia is evident at the earliest stage of NAFLD in ob/ob mice, which supports the concept that obesity-related metabolic abnormalities, rather than cirrhosis, initiate the hepatic neoplastic process during obesity. PMID- 11431336 TI - Tubulin detyrosination is a frequent occurrence in breast cancers of poor prognosis. AB - Tubulin, the dimeric subunit of microtubules, is a major cell protein that is centrally involved in cell division. Tubulin is subject to specific enzymatic posttranslational modifications including cyclic tyrosine removal and addition at the COOH terminus of the alpha-subunit. Tubulin is normally extensively tyrosinated in cycling cells. However, we have previously shown that detyrosinated tubulin accumulates in cancer cells during tumor progression in nude mice. Tubulin detyrosination, resulting from suppression of tubulin tyrosine ligase and the resulting unbalanced activity of tubulin-carboxypeptidase, apparently represents a strong selective advantage for cancer cells. We have now analyzed the occurrence and significance of tubulin detyrosination in human breast tumors. We studied a total of 134 breast cancer tumors from patients with or without known complications over a follow-up period of 31 +/- 10 months. The mean age of the patients at the time of diagnosis was 57 years. For each patient, detailed data concerning the histology and extension of the tumor were available. Tumor cells containing detyrosinated tubulin were visualized by immunohistochemical staining of paraffin-embedded tissue sections. Cancer cells with detyrosinated tubulin were observed in 53% of the tumors and were predominant in 19.4% of the tumors. Tubulin detyrosination correlated to a high degree of significance (P < 0.001) with a high Scarf-Bloom-Richardson (SBR) grade, a known marker of tumor aggressiveness. Among SBR grade 1 tumors, 3.8% were strongly positive for tubulin detyrosination compared with 65.4% of the SBR grade 3 tumors. The SBR component showing the strongest correlation with tubulin detyrosination was the mitotic score. In the entire patient population, neither the SBR grade nor the detyrosination index had significant prognostic value (P = 0.11, P = 0.27, respectively), whereas a combined index was significantly correlated with the clinical outcome (P = 0.02). A preliminary subgroup analysis indicated that tubulin detyrosination may define high- and low- risk groups in breast cancer tumors with an SBR grade of 2. Our study shows that tubulin detyrosination is a frequent occurrence in breast cancer, easy to detect, and linked to tumor aggressiveness. PMID- 11431337 TI - Increased level of stromal cell-derived factor-1 mRNA in peripheral blood mononuclear cells from children with AIDS-related lymphoma. AB - A common polymorphism in the 3' untranslated region of the stromal cell-derived factor 1 (also called pre-B-cell-stimulating factor) beta gene transcript, termed SDF1-3'A, has been associated with an increased risk of non-Hodgkin's lymphoma (NHL) in HIV-1-infected, but not in uninfected, individuals. Because the gene variation is located within the 3' untranslated region, the SDF1-3'A may influence the abundance of SDF-1 mRNA, possibly up-regulating the chemokine expression especially in the presence of HIV-1. In the current study, we investigated the levels of SDF-1 mRNA in peripheral blood mononuclear cells and HIV-1 viral load in 84 HIV-1-infected children (0.7 to 18 years of age; median, 5.8), including 12 children who developed NHL during their illnesses (AIDS-NHL group; 8 with SDF1-3'A, 4 with SDF1-wild-type). High level SDF-1 expression was observed in 15 of 34 children with SDF1-3'A as compared with 10 of 50 with wild type (P < 0.03). More notably, the children with AIDS-NHL had significantly elevated levels of SDF-1 mRNA in peripheral blood mononuclear cells, obtained at the time of presentation in 10 children and 8.5 to 19.4 months before (median, 15 months) in 7 children, as compared with the children in the non-NHL group (P < 0.00001). The amounts of cell-associated HIV-1 DNA and singly spliced HIV-1 mRNA were significantly greater in children with AIDS-NHL than those with non-NHL AIDS (P = 0.0052 and 0.011, respectively; stratified by antiretroviral treatment regimen), whereas their serum HIV-1 RNA levels were comparable. Overexpression of SDF-1 and aberrant HIV-1 expression in circulating lymphocytes appear to be linked to the development of AIDS-lymphoma. Additional studies are required to determine whether excessive SDF-1, together with virally encoded factors, is directly involved in the pathogenesis of AIDS-lymphoma. PMID- 11431338 TI - Conversion from a paracrine to an autocrine mechanism of androgen-stimulated growth during malignant transformation of prostatic epithelial cells. AB - Normal adult prostate epithelium of both human and rat origin was transplanted with Matrigel into intact or androgen-ablated (i.e., castrated) nude mice. Within these transplants, an influx of mouse mesenchymal cells was one of the earliest events to occur resulting in the development of a collar of smooth muscle cells and fibroblasts surrounding the transplanted epithelium. A subset of these surrounding stromal cells express androgen receptor (AR). The surrounded transplanted epithelium initially expresses high molecular weight cytokeratins characteristic of prostatic basal cells and AR. In both intact and androgen ablated hosts, this epithelium subsequently develops a patent lumen producing a rudimentary glandular acini. Only in the nonablated hosts, however, do these rudimentary acini undergo a further proliferative growth phase, as determined by Ki67 immunocytochemical stainings and the development of a low molecular weight cytokeratin positive layer of luminal (i.e., secretory) epithelial cells. Because AR is expressed in both the donor epithelium and host (i.e., mouse) stromal cells, this androgen-stimulated growth response could involve either autocrine pathways initiated within donor normal adult epithelial cells themselves or paracrine pathways initiated within the AR-expressing subset of mouse stromal cells. To resolve this issue, mice carrying the testicular feminized mutation in the X-linked AR gene were cross-bred to AR-wt nude mice to produce AR-null nude male mice. None of the cells in these AR-null nude male mice express functional AR protein. Therefore, these animals can be used to prevent any possibility of host stromal cell paracrine involvement in initiating an androgen-stimulated growth response when normal adult or malignant prostatic epithelial cells are transplanted into these null hosts. In these AR-null nude male mice, the androgen stimulated growth of normal adult prostatic epithelial cells did not occur (i.e., androgen-induced growth response of normal prostatic epithelial cells requires stromal cell paracrine involvement). In contrast, using four different prostatic cancer models (i.e., human PC-82, human LNCaP, human LAPC-4, and rat R3327G), the androgen-stimulated growth of prostatic cancer cells occurred identically in both AR-null and AR-wt nude male mice (i.e., a direct autocrine mechanism is responsible for androgen-stimulated growth of malignant prostatic epithelial cells). In summary, a fundamental change in the mechanism for androgen-stimulated growth occurs during the transformation from normal to malignant prostatic epithelial cells. PMID- 11431339 TI - Soy diets containing varying amounts of genistein stimulate growth of estrogen dependent (MCF-7) tumors in a dose-dependent manner. AB - We have demonstrated that the isoflavone, genistein, stimulates growth of estrogen-dependent human breast cancer (MCF-7) cells in vivo (C. Y. Hsieh et al., Cancer Res., 58: 3833-3838, 1998). The isoflavones are a group of phytoestrogens that are present in high concentrations in soy. Whether consumption of genistein from soy protein will have similar effects on estrogen-dependent tumor growth as pure genistein has not been investigated in the athymic mouse tumor implant model. Depending on processing, soy protein isolates vary widely in concentrations of genistein. We hypothesize that soy isolates containing different concentrations of genistein will stimulate the growth of estrogen dependent cells in vivo in a dose-dependent manner. To test this hypothesis we conducted experiments in which these soy protein isolates were fed to athymic mice implanted s.c. with estrogen-dependent tumors. Genistein content (aglycone equivalent) of the soy isolate diets were 15, 150, or 300 ppm. Positive (with 17beta-estradiol pellet implant) and negative (no 17beta-estradiol) control groups received casein-based (isoflavone-free) diets. Tumor size was measured weekly. At completion of the study animals were killed and tumors collected for evaluation of cellular proliferation and estrogen-dependent gene expression. Incorporation of bromodeoxyuridine into cellular DNA was used as an indicator of cell proliferation, and pS2 mRNA was used as an estrogen-responsive gene. Soy protein diets containing varying amounts of genistein increased estrogen dependent tumor growth in a dose-dependent manner. Cell proliferation was greatest in tumors of animals given estrogen or dietary genistein (150 and 300 ppm). Expression of pS2 was increased in tumors from animals consuming dietary genistein (150 and 300 ppm). Here we present new information that soy protein isolates containing increasing concentrations of genistein stimulate the growth of estrogen-dependent breast cancer cells in vivo in a dose-dependent manner. PMID- 11431340 TI - Association of NAT1 and NAT2 polymorphisms to urinary bladder cancer: significantly reduced risk in subjects with NAT1*10. AB - The role of hereditary polymorphisms of the arylamine N-acetyltransferase 1 (NAT1) gene in the etiology of urinary bladder cancer is controversial. NAT1 is expressed in the urothelium and may O-acetylate hydroxyl amines, particularly in subjects with low NAT2 activity. Thus, NAT1 polymorphisms may affect the individual bladder cancer risk by interacting with environmental factors (smoking and occupational risks) and by interacting with the NAT2 gene. We studied the frequencies of the NAT1 haplotypes *3, *4, *10, *11, *14, *15, *17, and *22 in 425 German bladder cancer patients and 343 controls by PCR-RFLP. NAT1*10 allelic frequency was lower in bladder cancer patients (15.1%) compared with controls (20.4%; P = 0.012). Genotypes that included NAT1*10 were significantly less frequent among the cases (odds ratio adjusted for age, gender, and smoking, 0.65; 95% confidence interval, 0.46-0.91; P = 0.013). Two subtypes of NAT1*11 were detected: *11A (-344T, -40T, 445A, 459A, 640G, and 1095A) and *11C (-344T, -40T, 459A, 640G, and 1095A). The allele frequency of NAT1*11 was 4.3% in the cases versus 3.9% in the controls. The rare low-active NAT1*14A was overrepresented in the cases (P = 0.026). With regard to the NAT2 genotype, our data showed: (a) a partial linkage of NAT1*10 to NAT2*4; (b) a clear underrepresentation of NAT1*10 genotypes among rapid NAT2 genotypes in the cases studied (odds ratio, 0.39; 95% confidence interval, 0.22-0.68; P = 0.001), and (c) a gene-gene-environment interaction. NAT2*slow/NAT1*4 genotype combinations with a history of occupational exposure were 5.96 (2.96-12.0) times more frequent in cancer cases than in controls without risk occupation (P < 0.0001). Hence, our data suggest that individuals provided with NAT2*4 and NAT1*10 are at a significantly lower risk for bladder cancer, particularly when exposed to environmental risk factors. PMID- 11431341 TI - Inhibition of fibroblast growth factor-2-induced vascular tumor formation by the acyclic nucleoside phosphonate cidofovir. AB - Cidofovir [(S)-HPMPC; (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine] is an antiviral drug that has been approved for the treatment of cytomegalovirus retinitis in AIDS patients. Cidofovir also possesses potent inhibitory activity against various human papillomavirus-induced tumors in animal models and patients. In addition, cidofovir inhibits the development of murine polyomavirus induced hemangiomas in rats by an as-yet-uncharacterized, antivirus-independent mechanism. Here we report the inhibitory effect of cidofovir on the development of virus-independent vascular tumors originated by basic fibroblast growth factor (FGF2)-overexpressing endothelial cells (FGF2-T-MAE cells). In vitro, cidofovir was cytostatic to FGF2-T-MAE cells at a 50% cytostatic concentration of 6.7 microg/ml. Cidofovir concentrations >25 microg/ml resulted in cytotoxicity because of induction of apoptosis. Cidofovir did not affect FGF2-T-MAE cell sprouting in three-dimensional fibrin gel and morphogenesis on Matrigel at noncytotoxic concentrations. In vivo, cidofovir (100 microg/egg) completely suppressed hemangioma formation on the chick chorioallantoic membrane (CAM) induced by intra-allantoic injection of FGF2-T-MAE cells, without affecting the formation of normal CAM vessels. Accordingly, cidofovir applied locally at 200 microg/disc, reduced neovascularization on the CAM by only 35%. Intratumoral or systemic administration of cidofovir caused a significant inhibition of the growth of s.c., i.p., or intracerebral FGF2-T-MAE xenografts in nude mice and severe combined immunodeficient mice. Drug-induced apoptosis was observed in FGF2 T-MAE tumors as soon as 2 days after the beginning of treatment. In conclusion, cidofovir appears to inhibit the growth of endothelium-derived tumors via induction of apoptosis without exerting a direct antiangiogenic activity. PMID- 11431342 TI - Zinc-65 imaging of rat brain tumors. AB - The uptake of zinc, an essential nutrient, is critical for cell proliferation. On the basis of the idea that zinc uptake can be an index of viability in proliferating cells, tumor imaging with (65)Zn was performed using autoradiography. After s.c. implantation of ascites hepatoma (AH7974F) cells into the dorsum, 1 h after i.v. injection of (65)ZnCl(2), (65)Zn uptake in the tumor was higher than in the brain tissue but lower than in the liver, which suggests that brain tumors can be positively imaged with (65)Zn. After implantation of AH7974F cells into the periaqueductal gray, 1 h after i.v. injection of (65)ZnCl(2), (65)Zn uptake in the tumor was approximately 10 times higher than in other brain regions. After implantation of C6 glioma cells into the hippocampus, (65)Zn uptake in the tumor was also much higher than in other brain regions. The present findings demonstrate that brain tumors can be imaged with radioactive zinc. To compare brain tumor imaging with (65)Zn with that of [(18)F]fluorodeoxyglucose (FDG), which is widely used for the diagnosis of brain tumors, (14)C-FDG imaging of the C6 glioma was performed in the same manner. (14)C-FDG uptake in the tumor was approximately 1.5 times higher than in the contralateral region in which (14)C-FDG uptake was relatively high. It is likely that zinc uptake is more specific for brain tumors than is FDG uptake, which suggests that there is great potential for the use of (69m)Zn, a short half-life gamma emitter, in the diagnosis of brain tumors. PMID- 11431343 TI - Lowering the isoelectric point of the Fv portion of recombinant immunotoxins leads to decreased nonspecific animal toxicity without affecting antitumor activity. AB - Recombinant immunotoxins are genetically engineered proteins in which the Fv portion of an antibody is fused to a toxin. Our laboratory uses a 38-kDa form of Pseudomonas exotoxin A termed PE38 for this purpose. Clinical studies with immunotoxins targeting CD25 and CD22 have shown that dose-limiting side effects are attributable to liver damage and other inflammatory toxicities. We recently showed that mutating exposed surface neutral residues to acidic residues in the framework region of the Fv portion of an immunotoxin targeting CD25 [anti Tac(scFv)-PE38] lowered its isoelectric point (pI) and decreased its toxicity in mice without impairing its cytotoxic or antitumor activities. We have now extended these studies and made mutations that change basic residues to neutral or acidic residues. Initially the pI of the mutant Fv (M1) of anti-Tac(scFv)-PE38 was decreased further. Subsequently, mutations were made in two other immunotoxins, SS1(dsFv)-PE38 targeting ovarian cancer and B3(dsFv)-PE38 targeting colon and breast cancers. We have found that all these mutant molecules fully retained specific target cell cytotoxicity and antitumor activity but were considerably less toxic to mice. Therefore, lowering the pI of the Fv may be a general approach to diminish the nonspecific toxicity of recombinant immunotoxins and other Fv fusion proteins without losing antitumor activity. PMID- 11431344 TI - Sensitization of human tumor cells to CPT-11 via adenoviral-mediated delivery of a rabbit liver carboxylesterase. AB - Irinotecan, 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin (CPT-11) is activated by carboxylesterases (CE) to yield the potent topoisomerase I inhibitor, SN-38. We have demonstrated previously that a rabbit liver CE is approximately 100-1000-fold more efficient at drug activation than a highly homologous human CE. In an attempt to use rabbit CE expression in combination with CPT-11 for gene therapy approaches for the treatment of cancer, we have developed an adenoviral vector expressing this intracellular CE. After transduction, this virus produces very high levels of CE activity in a panel of human tumor cell lines and results in marked sensitization to CPT-11 of all of the transduced cells. Reductions in IC(50) values for this drug ranged from 11 127-fold. Additionally, comparison with an adenovirus expressing a secreted form of the rabbit CE indicated that a collateral effect could be achieved with reductions in the IC(50) values ranging from 4-19-fold. These data suggest that the described reagents may be suitable for use in vivo in a viral-directed enzyme prodrug therapy approach using CPT-11. PMID- 11431345 TI - A virus-directed enzyme prodrug therapy approach to purging neuroblastoma cells from hematopoietic cells using adenovirus encoding rabbit carboxylesterase and CPT-11. AB - Tumor cells that contaminate hematopoietic cell preparations contribute to the relapse of neuroblastoma patients who receive autologous stem cell rescue as a component of therapy. Therefore, effective purging methods are needed. This study details in vitro experiments to develop a viral-directed enzyme prodrug purging method that specifically targets neuroblastoma cells. The approach uses an adenovirus to deliver the cDNA encoding a rabbit liver carboxylesterase that efficiently activates the prodrug irinotecan,7-ethyl-10-[4-(1-piperidino)-1 piperidino]carbonyloxycamptothecin (CPT-11). The data show that an adenoviral multiplicity of infection of 50 transduces 100% of cultured neuroblastoma cells and primary tumor cells, irrespective of the level of tumor cell line contamination. Exposure of neuroblastoma cell lines or of mixtures of these cell lines with CD34(+) cells at a ratio of 10:90 to replication-deficient AdRSVrCE for 24 h and subsequent exposure of cells to 1-5 microM CPT-11 for 4 h increased the toxicity of CPT-11 to three neuroblastoma cell lines (SJNB-1, NB-1691, and SK N-SH) from approximately 20-50-fold and eradicated their clonogenic potential. Also, after "purging," RNA for neuroblastoma cell markers (tyrosine hydroxylase, synaptophysin, and N-MYC) was undetectable by reverse transcription-PCR. In contrast, the purging protocol did not affect the number or type of colonies formed by CD34(+) cells in an in vitro progenitor cell assay. No bystander effect on CD34(+) cells was observed. The method described is being investigated for its potential clinical utility, particularly its efficacy for use with patients having relatively high tumor burdens, because no published methods have been shown to be efficacious when the tumor burden exceeds 1%. PMID- 11431346 TI - Acquired resistance to the antitumor effect of epidermal growth factor receptor blocking antibodies in vivo: a role for altered tumor angiogenesis. AB - Inhibitors of epidermal growth factor receptor (EGFR) signaling are among the novel drugs showing great promise for cancer treatment in the clinic. However, the possibility of acquired resistance to such drugs because of tumor cell genetic instabilities has not yet been explored. Here we report the experimental derivation and properties of such cell variants obtained from recurrent tumor xenografts of the human A431 squamous cell carcinoma, after two consecutive cycles of therapy with one of three different anti-EGFR monoclonal antibodies: mR3, hR3, or C225. Initial response to a 2-week period of treatment was generally total tumor regression and was not significantly different among the three antibody groups. However, tumors often reappeared at the site of inoculation, generally after prolonged latency periods, and most of the tumors became refractory to a second round of therapy. Cell lines established from such resistant tumors retained high EGFR expression, normal sensitivity to anti-EGFR antibody or ligand, and unaltered growth rate when compared with the parental line in vitro. In contrast, the A431 cell variants exhibited an accelerated growth rate and a significantly attenuated response to anti-EGFR antibodies in vivo relative to the parental line. Because of the reported suppressive effect of EGFR inhibitors on vascular endothelial growth factor (VEGF) expression, and the demonstrated role of VEGF in the angiogenesis and growth of A431 tumor xenografts, relative VEGF expression was examined. Five of six resistant variants expressed increased levels of VEGF, which paralleled an increase in both angiogenic potential in vitro and tumor angiogenesis in vivo. In addition, elevated expression of VEGF in variants of A431 cells obtained by gene transfection rendered the cells significantly resistant to anti-EGFR antibodies in vivo. Taken together, the results suggest that, at least in the A431 system, variants displaying acquired resistance to anti-EGFR antibodies can emerge in vivo and can do so, at least in part, by mechanisms involving the selection of tumor cell subpopulations with increased angiogenic potential. PMID- 11431347 TI - N-(4-hydroxyphenyl)retinamide elevates ceramide in neuroblastoma cell lines by coordinate activation of serine palmitoyltransferase and ceramide synthase. AB - The retinoid N-(4-hydroxyphenyl)retinamide (4-HPR; fenretinide) is cytotoxic to a variety of cancer cell lines, and we previously showed an association between ceramide generation and 4-HPR cytotoxicity for neuroblastoma cell lines (B. J. Maurer et al., J. Natl. Cancer Inst. (Bethesda), 91: 1138-1146, 1999). Here we determine whether the increased ceramide mediated by 4-HPR in the CHLA-90 human neuroblastoma cell line results from de novo ceramide synthesis. Treatment of CHLA-90 with 4-HPR for 2 h, in the presence of [(3)H]palmitic acid, caused sequential formation of [(3)H]sphinganine (220% over control) and [(3)H]ceramide (160% over control), with sphinganine returning to baseline at 4 h, and ceramide continuing to increase (215% over control). 4-HPR treatment did not accelerate cellular decay of sphingomyelin. Preincubation of cells with either L cycloserine, an inhibitor of serine palmitoyltransferase (SPT), or fumonisin B(1), an inhibitor of ceramide synthase, retarded ceramide formation in response to 4-HPR treatment, although sphinganine was still generated when 4-HPR and FB(1) were present. Data from in vitro enzyme assays using microsomes showed that preexposure of intact cells to 4-HPR resulted in a time (175% over control; 6 h)- and dose-dependent increase (173% over control; 10 microM) in SPT activity as well as a time (265% over control)- and dose-dependent increase (215% above control; 10 microM) in ceramide synthase activity. Our results show that 4-HPR mediated ceramide generation is derived from the de novo synthetic pathway by coordinate activation of SPT and ceramide synthase. Knowledge of these biochemical events is of utility when downstream modulators of ceramide metabolism are used to heighten the cytotoxic response to chemotherapy. PMID- 11431348 TI - Pharmacological inhibitors of the mitogen-activated protein kinase (MAPK) kinase/MAPK cascade interact synergistically with UCN-01 to induce mitochondrial dysfunction and apoptosis in human leukemia cells. AB - Interactions between the checkpoint abrogator UCN-01 and several pharmacological inhibitors of the mitogen-activated protein kinase (MAPK) kinase (MEK)/MAPK pathway have been examined in a variety of human leukemia cell lines. Exposure of U937 monocytic leukemia cells to a marginally toxic concentration of UCN-01 (e.g., 150 nM) for 18 h resulted in phosphorylation/activation of p42/44 MAPK. Coadministration of the MEK inhibitor PD184352 (10 microM) blocked UCN-01-induced MAPK activation and was accompanied by marked mitochondrial damage (e.g., cytochrome c release and loss of DeltaPsi(m)), caspase activation, DNA fragmentation, and apoptosis. Similar interactions were noted in the case of other MEK inhibitors (e.g., PD98059; U0126) as well as in multiple other leukemia cell types (e.g., HL-60, Jurkat, CCRF-CEM, and Raji). Coadministration of PD184352 and UCN-01 resulted in reduced binding of the cdc25C phosphatase to 14-3 3 proteins, enhanced dephosphorylation/activation of p34(cdc2), and diminished phosphorylation of cyclic AMP-responsive element binding protein. The ability of UCN-01, when combined with PD184352, to antagonize cdc25C/14-3-3 protein binding, promote dephosphorylation of p34(cdc2), and potentiate apoptosis was mimicked by the ataxia telangectasia mutation inhibitor caffeine. In contrast, cotreatment of cells with UCN-01 and PD184352 did not substantially increase c-Jun-NH(2) terminal kinase activation nor did it alter expression of Bcl-2, Bcl-x(L), Bax, or X-inhibitor of apoptosis. However, coexposure of U937 cells to UCN-01 and PD184352 induced a marked increase in p38 MAPK activation. Moreover, SB203580, which inhibits multiple kinases including p38 MAPK, partially antagonized cell death. Lastly, although UCN-01 +/- PD184352 did not induce p21(CIP1), stable expression of a p21(CIP1) antisense construct significantly increased susceptibility to this drug combination. Together, these findings indicate that exposure of leukemic cells to UCN-01 leads to activation of the MAPK cascade and that interruption of this process by MEK inhibition triggers perturbations in several signaling and cell cycle regulatory pathways that culminate in mitochondrial injury, caspase activation, and apoptosis. They also raise the possibility that disrupting multiple signaling pathways, e.g., by combining UCN 01 with MEK inhibitors, may represent a novel antileukemic strategy. PMID- 11431349 TI - Retrovirus-mediated expression of the base excision repair proteins, formamidopyrimidine DNA glycosylase or human oxoguanine DNA glycosylase, protects hematopoietic cells from N,N',N"-triethylenethiophosphoramide (thioTEPA)-induced toxicity in vitro and in vivo. AB - Modulation of DNA damage repair activity could lead to new approaches to reduce cytotoxic side effects of chemotherapy. N,N',N"-Triethylenethiophosphoramide (thioTEPA) induces the formation of amino-ethyl adducts of guanine, resulting in imidazole ring opening [formamidopyrimidine (Fapy)] and is associated with significant myelosuppression in dose-intensive therapies. In Escherichia coli, Fapy lesions are repaired by the Fapy-DNA glycosylase (Fpg) protein. We hypothesized that the expression of the Fpg could increase resistance of hematopoietic cells to thioTEPA-induced cytotoxicity. Expression of Fpg in bone marrow (BM) cells via a retrovirus vector was associated with demonstrable 8 oxodeoxyguanosine DNA glycosylase activity. BM cells were infected with a recombinant retrovirus, SF91, containing the Fpg gene and expressing the enhanced green fluorescence protein (EGFP) via an internal ribosomal entry site element. Control mice received BM transduced with the backbone containing IRES-EGFP alone. Fpg-transduced and GFP+ BM hematopoietic cells were resistant in vitro to thioTEPA at multiple concentrations. Mice transplanted with transduced cells were treated with four doses of thioTEPA (10 mg/kg) given over 7 weeks. Despite low transduction efficiency, peripheral blood leukocytes, hemoglobin, and platelet counts of thioTEPA-treated Fpg mice were significantly higher than treated control mice (P < 0.05). In addition, after treatment, the BM, spleen, and thymic cellularity as well as the number of GFP+ progenitor cells in the BM of treated mice were significantly higher than those of control group. Selection of Fpg transduced cells in vivo was demonstrated by an increase in the mean fluorescence intensity of peripheral mononuclear cells of Fpg mice compared with pretreatment value. In addition, a significant increase in the EGFP-bright cells was demonstrated, suggesting preferential survival of high-expressing hematopoietic cells. Similar results were demonstrated in vitro with primary BM expressing the human functional counterpart of Fpg, OGG1. These results show that expression of the Fpg or hOGG1 protein protects hematopoietic cells from thioTEPA-induced DNA damage and suggest that a high level of expression of these repair proteins is required to establish resistance to this drug. Expression of Fpg and/or OGG1 may provide an novel approach to preventing thioTEPA-induced toxicity of primary hematopoietic cells. PMID- 11431350 TI - Immunosuppressive effects of (131)I-anti-CD45 antibody in unsensitized and donor antigen-presensitized H2-matched, minor antigen-mismatched murine transplant models. AB - Iodine-131-labeled anti-CD45 antibody has been added to conventional hematopoietic stem cell transplant preparative regimens to deliver targeted radiation to hematopoietic tissues, with the goal of decreasing relapse rates without increasing toxicity. However, higher radiation doses could be delivered to leukemia cells by antibody if the systemic therapy were decreased or eliminated. To examine the ability of (131)I-anti-CD45 antibody to provide sufficient immunosuppression for transplantation across allogeneic barriers, T cell-depleted BALB.B marrow was transplanted into H2-compatible B6-Ly5(a) mice after (131)I-30F11 (rat antimurine CD45) antibody with or without varying dose levels of total body irradiation (TBI). Groups of five or six recipient mice per (131)I or TBI dose level per experiment were given tail vein injections of 100 microg of (131)I-labeled 30F11 antibody 4 days before marrow infusion, with or without TBI on day 0. Engraftment, defined as > or =50% donor B cells at 3 months posttransplant, was determined by two-color flow cytometric analysis of peripheral blood granulocytes, T cells, and B cells using antibodies specific for donor and host CD45 allotypes and for CD3. Donor engraftment of > or =80% recipient mice was achieved with either 8 Gy of TBI or 0.75 mCi of (131)I-30F11 antibody, which delivers an estimated 26 Gy to bone marrow. Subsequent experiments determined the dose of TBI alone or TBI plus 0.75 mCi of (131)I-30F11 antibody necessary for engraftment in recipient mice that had been presensitized to donor antigens before transplant, a setting requiring more stringent immunosuppression. Engraftment was seen in > or =80% of presensitized recipients surviving after 14-16 Gy of TBI or 12-14 Gy of TBI and 0.75 mCi of (131)I-30F11 antibody. However, only 28 of 69 (41%) presensitized mice receiving 10-16 Gy of TBI alone survived, presumably because of rejection of donor marrow and ablation of host hematopoiesis. In contrast, 29 of 35 (83%) presensitized mice receiving (131)I-30F11 antibody and 10-14 Gy of TBI survived, presumably because the additional immunosuppression provided by estimated radiation doses of 53 Gy to lymph nodes and 81 Gy to spleen from 0.75 mCi of (131)I-30F11 antibody permitted engraftment of donor marrow. These results suggest that targeted radiation delivered by (131)I-anti-CD45 antibody provides sufficient immunosuppression to replace an appreciable portion of the TBI dose used in matched sibling hematopoietic stem cell transplant. PMID- 11431351 TI - Proliferative activity of intratumoral CD8(+) T-lymphocytes as a prognostic factor in human renal cell carcinoma: clinicopathologic demonstration of antitumor immunity. AB - Tumor-infiltrating lymphocytes, particularly CD8(+) T cells, could be a manifestation of antitumor immunity. We clinicopathologically analyzed the biological significance of tumor-infiltrating lymphocytes in 221 patients with renal cell carcinoma without preoperative treatments. More abundant infiltration of tumor tissue not only by CD8(+) but also CD4(+) T cells was associated with shorter survival of the patients, because of the positive correlation between the number of lymphocytes and representative tumor grade factors. This suggests that immune cell reactions are more pronounced as the tumor grade/biological malignancy progresses, probably because of increased antigenicity of tumor cells. We next analyzed the proliferative activity of CD8(+) T cells that infiltrated in tumor cell nests, which could also reflect antitumor immunity. Higher labeling index of Ki-67, a proliferation-associated antigen, among CD8(+) T cells in contact to tumor cells was associated with a longer survival by both uni- and multivariate analyses. Our data in human renal cell carcinoma suggest that infiltration of tumor tissue by T cells itself does not denote the efficacy of antitumor immunity because of its dependence on the biological malignancy of tumor cells, but infiltration of tumor tissue by CD8(+) T cells bearing more pronounced proliferative activity could reflect effective antitumor immunity. This concept would be important for future immunotherapy of human cancer. PMID- 11431352 TI - Rituximab inactivates signal transducer and activation of transcription 3 (STAT3) activity in B-non-Hodgkin's lymphoma through inhibition of the interleukin 10 autocrine/paracrine loop and results in down-regulation of Bcl-2 and sensitization to cytotoxic drugs. AB - Development of the chimeric mouse antihuman CD20 antibody, Rituximab, presented a notable advance in the treatment of patients with non-Hodgkin's lymphoma (NHL). Its use allowed the specific targeting of tumor B cells without the systemic toxicity of traditional therapies. The mechanisms by which Rituximab induces its antitumor activity are not fully understood. We have shown previously that Rituximab down-regulates Bcl-2 expression in some B-NHL cell lymphoma lines through an interleukin 10 (IL-10)-dependent autocrine loop, an effect that renders the resistant cells susceptible to chemotherapeutic drugs. The objective of this study was to delineate the signaling pathway by which Bcl-2 is controlled by Rituximab and IL-10. We hypothesized that the down-regulation of IL-10 by Rituximab decreases activation of the signal transducer and activator of transcription 3 (STAT3) protein, which in turn, is responsible for decreased levels of Bcl-2. We demonstrate by phosphoprotein immunoblotting and gel shift analyses that endogenous IL-10 induces activation of STAT3 in the 2F7 cell line. Furthermore, we show that Rituximab and anti-IL-10 antibody treatment decreases the ability of STAT3 to bind to its DNA binding site. The decrease in STAT3 activation by these treatments correlates with a decrease in Bcl-2 expression. Additionally, piceatannol, an inhibitor of STAT3 activation, down-regulates the expression of Bcl-2. Altogether, these results demonstrate that Bcl-2 expression is under the regulation of the STAT3 signaling pathway, which is regulated by endogenously secreted IL-10. Hence, Rituximab-induced down-regulation of IL-10 expression is responsible for the down-regulation of Bcl-2 and sensitization of NHL cells by therapeutic drugs. Furthermore, these findings support the notion that circulating IL-10 in vivo may control the resistance of NHL to drug-mediated cytotoxicity. PMID- 11431353 TI - Somatically mutated regions of immunoglobulin on human B-cell lymphomas code for peptides that bind to autologous major histocompatibility complex class I, providing a potential target for cytotoxic T cells. AB - Lymphoma-derived immunoglobulin idiotype (Id) is a well-characterized, tumor specific antigen on B-cell malignancies. Immunotherapy using lymphoma immunoglobulin can lead to clinical responses mostly associated with anti-Id antibody. We cloned the Id from B-cell lymphomas, sequenced them, and used bioinformatics to select autologous MHC class I binding peptides from somatically mutated regions of the lymphoma Id. Peptides from patients who were HLA-A1, HLA A2, HLA-A3, or HLA-A11 positive were analyzed in the T2 stabilization assay and a competitive peptide-binding assay. By both methods, approximately half of the peptides analyzed, regardless of HLA type, bound with intermediate or high affinity. Peptide binding affinity was similar to viral peptide sequences known to provide targets for cytotoxic T cells. Further investigation of lymphocyte responses to stimulation by autologous Id peptides versus Id peptides from other patients revealed that three of five patients in complete remission or with low volume, stable disease responded to self-peptides by IFN-gamma secretion greater than that seen with non-self peptides, whereas none of five patients with progressive disease responded to their own lymphoma Id. We have shown that mutated regions of lymphoma Id contain MHC class I binding peptides that are potential targets for cytotoxic T cells. Immunotherapy using the tumor-specific mutated regions from lymphoma Id avoids the need to break innate tolerance toward the germ-line protein sequences present on normal and malignant B cells. PMID- 11431354 TI - Combinatorial peptide libraries as an alternative approach to the identification of ligands for tumor-reactive cytolytic T lymphocytes. AB - The recent identification of molecularly defined human tumor antigens recognized by autologous CTLs has opened new opportunities for the development of antigen specific cancer vaccines. Despite extensive work, however, the number of CTL defined tumor antigens that are suitable targets for generic vaccination of cancer patients is still limited, mostly because of the painstaking and lengthy nature of the procedures currently used for their identification. A novel approach is based on the combined use of combinatorial peptide libraries in positional scanning format (positional scanning synthetic combinatorial peptide libraries, PS-SCLs) and tumor-reactive CTL clones. To validate this approach, we herein analyzed in detail the recognition of PS-SCLs by Melan-A-specific CTL clones. Our results indicate that, at least for some clones, most of the amino acids composing the native antigenic peptide can be identified through the use of PS-SCLs. Interestingly, this analysis also allowed the identification of peptide analogues with increased antigenic activity as well as agonist peptides containing multiple amino-acid substitutions. In addition, biometrical analysis of the data generated by PS-SCL screening allowed the identification of the native ligand in a public database. Overall, these data demonstrate the successful use of PS-SCLs for the identification and optimization of tumor associated CTL epitopes. PMID- 11431355 TI - Identification of T helper epitopes from prostatic acid phosphatase. AB - Helper T cells (Th cells) play a central role in the initiation and maintenance of immune responses, including antitumor immunity. The ability of Th cells in murine models to maintain and enhance the cytolytic efficacy of CD8+ CTLs has led to a renewed interest in identifying human tumor antigens recognized by Th cells. Prostatic acid phosphatase (PAP) is a prostate cancer-associated tumor antigen. A rodent model has demonstrated that PAP-specific CTLs can induce destructive prostatitis. Human MHC class I epitopes derived from PAP have been identified previously, and peptide-specific CTLs have been shown to be able to lyse an MHC restricted prostate cancer cell line. In the current study, we sought to identify Th epitopes derived from PAP that might be used to elicit PAP-specific Th responses, ultimately in the context of human vaccines targeting PAP. Using peripheral blood mononuclear cells (PBMCs) from subjects with and without PAP specific Th responses, we screened a panel of 10 potential peptide epitopes for peptide-specific T-cell proliferation. Four peptides, p81-95, p199-213, p228-242, and p308-322, were identified for which peptide-specific T-cell proliferation occurred in the majority of patient PBMC samples that also exhibited PAP-specific T-cell proliferation. PBMCs from patients with prostate cancer and without PAP specific Th immunity were then cultured in vitro with these four peptides. Peptide-specific T-cell lines could be generated from two of the four peptides, p199-213 and p228-242, that also proliferated in response to PAP protein stimulation. The ability of these two peptides to elicit PAP-specific Th responses suggests that they represent naturally processed PAP-specific MHC class II epitopes. PMID- 11431356 TI - Identification of gene expression profiles that predict the aggressive behavior of breast cancer cells. AB - With the goal of identifying genes that have an expression pattern that can facilitate the diagnosis of primary breast cancers (BCs) as well as the discovery of novel drug leads for BC treatment, we used cDNA hybridization arrays to analyze the gene expression profiles (GEPs) of nine weakly invasive and four highly invasive BC cell lines. Differences in gene expression between weakly and highly invasive BC cells were identified that enabled the definition of consensus GEPs for each invasive phenotype. To determine whether the consensus GEPs, comprising 24 genes, could be used to predict the aggressiveness of previously uncharacterized cells, gene expression levels and comparative invasive and migratory characteristics of nine additional human mammary epithelial cell strains/lines were determined. The results demonstrated that the GEP of a cell line is predictive of its invasive and migratory behavior, as manifest by the morphology of its colonies when cultured on a matrix of basement membrane constituents (i.e., Matrigel). We found that the expression of keratin 19 was consistently elevated in the less aggressive BC cell lines and that vimentin and fos-related antigen-1 (FRA-1) were consistently overexpressed in the more highly aggressive BC cells. Moreover, even without these three genes, the GEP of a cell line still accurately predicted the aggressiveness of the BC cell, indicating that the expression pattern of multiple genes may be used as BC prognosticators because single markers often fail to be predictive in clinical specimens. PMID- 11431357 TI - Constitutional genomic instability with inversions, duplications, and amplifications in 9p23-24 in BRCA2 mutation carriers. AB - Germ-line mutations of the BRCA2 gene (13q12-13) account for a large proportion of familial breast cancer cases in females and the majority of familial breast cancers in males. Recent studies provide evidence for a role of the BRCA2 protein in the maintenance of genomic integrity by involvement in DNA repair and recombination. In pursuit of identifying in humans genetic damage resulting from mutated BRCA2, we have analyzed constitutional karyotypes of BRCA2 mutation carriers. The present study establishes that constitutional distal 9p rearrangements without obvious additional gross chromosomal alterations are a recurrent feature of independently ascertained families. From our cytogenetic analyses we have no indication of additional gross rearrangements, but we cannot exclude more subtle recombinations in other genomic regions. We also show that the topography of the 9p rearrangements can differ among family members, even within an individual that can have cell populations with different 9p rearrangements. Collectively these results raise point to an association of mutant BRCA2 with genomic instability and gene alteration in 9p23-24 in at least a subset of BRCA2 mutation carriers. PMID- 11431358 TI - Genetic evidence for early divergence of small functioning and nonfunctioning endocrine pancreatic tumors: gain of 9Q34 is an early event in insulinomas. AB - The malignant potential among endocrine pancreatic tumors (EPTs) varies greatly and can frequently not be predicted using histopathological parameters. Thus, molecular markers that can predict the biological behavior of EPTs are required. In a previous comparative genomic hybridization study, we observed marked genetic differences between the various EPT subtypes and a correlation between losses of 3p and 6 and gains of 14q and Xq and metastatic disease. To search for genetic alterations that play a role during early tumor development, we have studied 38 small (< or =2 cm) EPTs, including 24 insulinomas and 10 nonfunctioning EPTs. Small EPTs are usually classified as clinically benign tumors in the absence of histological signs of malignancy. Using comparative genomic hybridization, we identified chromosomal aberrations in 27 EPTs (mean, 4.1). Interestingly, the number of gains differed strongly between nonfunctioning and functioning EPTs (3.4 versus 1.5, respectively; P = 0.0526), as did the number of aberrations in the benign (n = 30) and malignant (n = 8) tumors (3 versus 8.4, respectively; P = 0.0022). In the insulinomas, 9q gain (common region of involvement: 9q34) was most common (50%) and in nonfunctioning EPTs, gain of 4p was most common (40%). Most frequent losses in insulinomas involved 1p (20.8%), 1q, 4q, 11q, Xq, and Y (all 16.7%) and in nonfunctioning EPTs, 6q. Losses of 3pq and 6q and gains of 17pq and 20q proved to be strongly associated with malignant behavior in all of the small EPTs (P < or = 0.0219). Our results demonstrate marked genetic differences between small functioning and nonfunctioning EPTs, indicating that these subtypes evolve along different genetic pathways. In addition, our study endorses the importance of chromosomes 3 and 6q losses to discriminate EPTs with a malignant behavior from benign ones. PMID- 11431359 TI - Role of the hMLH1 DNA mismatch repair protein in fluoropyrimidine-mediated cell death and cell cycle responses. AB - DNA mismatch repair (MMR) is an efficient system for the detection and repair of mismatched and unpaired bases in DNA. Deficiencies in MMR are commonly found in both hereditary and sporadic colorectal cancers, as well as in cancers of other tissues. Because fluorinated thymidine analogues (which through their actions might generate lesions recognizable by MMR) are widely used in the treatment of colorectal cancer, we investigated the role of MMR in cellular responses to 5 fluorouracil and 5-fluoro-2'-deoxyuridine (FdUrd). Human MLH1(-) and MMR deficient HCT116 colon cancer cells were 18-fold more resistant to 7.5 microM 5 fluorouracil (continuous treatment) and 17-fold more resistant to 7.5 microM FdUrd in clonogenic survival assays compared with genetically matched, MLH1(+) and MMR-proficient HCT116 3-6 cells. Likewise, murine MLH1(-) and MMR-deficient CT-5 cells were 3-fold more resistant to a 2-h pulse of 10 microM FdUrd than their MLH1(+) and MMR-proficient ME-10 counterparts. Decreased cytotoxicity in MMR-deficient cells after treatment with various methylating agents and other base analogues has been well reported and is believed to reflect a tolerance to DNA damage. Synchronized HCT116 3-6 cells treated with a low dose of FdUrd had a 2-fold greater G(2) cell cycle arrest compared with MMR-deficient HCT116 cells, and asynchronous ME-10 cells demonstrated a 4-fold greater G(2) arrest after FdUrd treatment compared with CT-5 cells. Enhanced G(2) arrest in MMR-proficient cells in response to other agents has been reported and is believed to allow time for DNA repair. G(2) cell cycle arrest as determined by propidium iodide staining was not a result of mitotic arrest, but rather a true G(2) arrest, as indicated by elevated cyclin B1 levels and a lack of staining with mitotic protein monoclonal antibody 2. Additionally, p53 and GADD45 levels were induced in FdUrd treated HCT116 3-6 cells. DNA double-strand break (DSB) formation was 2-fold higher in MMR-proficient HCT116 3-6 cells after FdUrd treatment, as determined by pulsed-field gel electrophoresis. The formation of DSBs was not the result of enhanced apoptosis in MMR-proficient cells. FdUrd-mediated cytotoxicity was caused by DNA-directed and not RNA-directed effects, because administration of excess thymidine (and not uridine) prevented cytotoxicity, cell cycle arrest, and DSB formation. hMLH1-dependent responses to fluoropyrimidine treatment, which may involve the action of p53 and the formation of DSBs, clearly have clinical relevance for the use of this class of drugs in the treatment of tumors with MMR deficiencies. PMID- 11431360 TI - Molecular mechanisms of transcriptional control of bcl-2 and c-myc in follicular and transformed lymphoma. AB - A synergistic interaction of Bcl-2 and c-Myc plays a role in lymphomagenesis in mice and in some patients as well. Progression of follicular lymphoma to a more aggressive lymphoma is seen in the majority of patients, and approximately 10% of the transformed lymphomas have a translocation of c-myc in addition to the translocation of bcl-2 found in the original follicular lymphoma. We investigated whether transcriptional deregulation of bcl-2 and c-myc could be examined in primary lymphoma cells by in vivo footprinting and in vitro protein-DNA binding studies. A matched pair of follicular and transformed lymphoma samples was examined. The transformed lymphoma had acquired a translocation of c-myc into the immunoglobulin heavy chain locus. High levels of bcl-2 expression were observed in both the follicular and transformed lymphomas, whereas the expression of c-myc was low in the follicular lymphoma and increased in the transformed lymphoma. In vivo footprint analysis revealed that a CRE site and a Cdx site in the bcl-2 promoter were occupied on the translocated alleles but not on the normal alleles in both the follicular and transformed lymphomas. Two nuclear factor kappaB sites were occupied on the translocated c-myc allele in the transformed lymphoma. Gel shift analysis revealed that these proteins bound to their respective sites in the bcl-2 or c-myc promoter. There was no evidence that the presence of one of the translocations in the immunoglobulin heavy chain locus influenced the expression of the other translocated gene. PMID- 11431361 TI - Hyaluronan synthase 3 overexpression promotes the growth of TSU prostate cancer cells. AB - Hyaluronan synthase 3 (HAS3) is responsible for the production of both secreted and cell-associated forms of hyaluronan and is the most active of the three isoforms of this enzyme in adults. In this study, the cDNA for human HAS3 was cloned and characterized. The open reading frame consisted of 1659 bp coding for 553 amino acids with a deduced molecular weight of about 63,000 and isoelectric pH of 8.70. The sequence of human HAS3 displayed a 53% identity to HAS1 and a 67% identity to HAS2. It also contained a signal peptide and six potential transmembrane domains, suggesting that it was associated with the plasma membrane. To evaluate the physiological role of human HAS3, expression vectors for this protein were transfected into TSU cells (a prostate cancer cell line), and the phenotypic changes in these cells were examined. The enhanced expression of hyaluronan in the transfected cells was demonstrated by dot blot analysis and ELISA. These cells were found to differ from their vector-transfected counterparts with respect to the following: (a) they grew at a faster rate in high (but not low) density cultures; (b) conditioned media from these cells stimulated the proliferation and migration of endothelial cells; (c) when placed on the chorioallantoic membrane of chicken embryos, these cells formed large, dispersed xenografts, whereas the control transfectants formed compact masses; and (d) when injected s.c. into nude mice, the xenografts formed by HAS3 transfectants were bigger than those formed by control transfectants. Histological examination of these xenografts revealed the presence of extracellular hyaluronan that could act as conduits for the diffusion of nutrients. In addition, they had a greater number of blood vessels. However, the HAS3-transfected TSU cells did not display increased metastatic properties as judged by their ability to form lung masses after i.v. injection. These results suggested that the HAS3-induced overexpression of hyaluronan enhanced tumor cell growth, extracellular matrix deposition, and angiogenesis but was not sufficient to induce metastatic behavior in TSU cells. PMID- 11431362 TI - Constitutive activation of hypoxia-inducible genes related to overexpression of hypoxia-inducible factor-1alpha in clear cell renal carcinomas. AB - The transcription factor hypoxia-inducible factor (HIF)-1 is an important mediator of hypoxic adaptation of tumor cells and controls several genes that have been implicated in tumor growth. Oxygen-dependent degradation of HIF-1alpha, the regulatory subunit, requires binding to the von Hippel Lindau (VHL) protein. Because functional inactivation of the VHL tumor suppressor gene occurs in up to 70% of clear cell renal carcinomas, we investigated whether this results in overexpression of HIF-1alpha and its target genes. Immunoblotting revealed increased expression of HIF-1alpha in 24 of 32 (75%) clear cell renal carcinomas but only 3 of 8 non-clear cell renal tumors. Somatic mutations of the VHL gene were detected only in clear cell renal carcinomas that overexpressed HIF-1alpha. None of the HIF-1alpha-negative tumors displayed a VHL mutation. The level of HIF 1alpha mRNA was not different between tumors and adjacent kidney tissue. Immunohistochemistry revealed distinct patterns of nuclear staining for HIF 1alpha, depending on histological type and overall abundance of HIF-1alpha. In those clear cell renal carcinomas that showed increased expression on immunoblots, HIF-1alpha was expressed in almost all cells. In the remaining clear cell and in non-clear cell tumors, staining was focal; these different patterns thus were compatible with genetic stabilization in contrast to microenvironmental stimulation of HIF-1alpha as the primary mechanism. The mRNA expression of two known target genes of HIF-1alpha, vascular endothelial growth factor and glucose transporter 1, increased progressively with increasing amounts of HIF-1alpha in tumor extracts. In addition, glucose transporter 1 protein levels correlated with HIF-1alpha abundance. In conclusion, the data provide in vivo evidence for a constitutive up-regulation of HIF-1alpha in the majority of clear cell renal carcinomas, which leads to more widespread accumulation of this transcription factor than hypoxic stimulation. These observations are most likely linked to functional inactivation of the VHL gene product. Increased expression of HIF 1alpha is associated with alterations in gene expression patterns that are likely to contribute to tumor phenotype and progression. PMID- 11431363 TI - Global analysis of gene expression in invasion by a lung cancer model. AB - Metastasis is a complicated multistep process that involves interactions between cancer cells and their surrounding microenvironments. Previously, we have established a series of lung adenocarcinoma cell lines with varying degrees of invasiveness. Tracheal graft assay confirmed that cell lines with higher in vitro invasiveness had greater in vivo invasive potential. In this study, we used these model cell lines to identify invasion-associated genes using cDNA microarray with colorimetric detection. A more invasive subline, CL 1-5-F 4, derived from metastatic lung tumor of severe combined immunodeficient mice inoculated with CL 1-5 cells, was combined with CL 1-0, CL 1-1, and CL 1-5 in cDNA microarray screening. cDNA microarray membranes, each containing 9600 nonredundant expressed sequence tag clones, were used to identify differentially expressed genes in these cell lines. For statistical analysis, self-organizing map algorithm was performed to identify the expression patterns. Positive correlation between gene expression levels and cell line invasiveness was found in 2.9% of the 9600 putative genes. On the other hand, negative correlation was found in 3.3% of the genes. The trends of expression of some of the genes were also confirmed by Northern hybridization and flow cytometry. Our data demonstrated that genes related to cell adhesion, motility, angiogenesis, signal transduction, and some other expressed sequence tag genes may play significant roles in the metastasis process. These results substantiate the model system with which one can identify invasion-associated genes by using cDNA microarray and cancer cell lines of different invasiveness. This technique may allow us to explore complex interactions between multiple genes that orchestrate the process of cancer metastasis. PMID- 11431364 TI - An intact overexpressed E-cadherin/alpha,beta-catenin axis characterizes the lymphovascular emboli of inflammatory breast carcinoma. AB - The step of intravasation (lymphovascular invasion), a rate-limiting step in metastasis, is greatly exaggerated in inflammatory breast carcinoma (IBC). Because nearly all human breast carcinoma cell lines grow as solitary nodules in nude/severe combined immunodeficient mice without manifesting lymphovascular invasion, this step has been difficult to study. We captured the essence of the IBC phenotype by establishing a unique human transplantable IBC xenograft, MARY X, which manifests florid lymphovascular emboli in severe combined immunodeficient/nude mice. Comparing MARY-X with common non-IBC cell lines/xenografts, we discovered an overexpressed and overfunctioning E cadherin/alpha,beta-catenin axis. In MARY-X, the E-cadherin and catenins were part of a structurally and functionally intact adhesion axis involving the actin cytoskeleton. In vitro, MARY-X grew as round compact spheroids with a cell density 5-10-fold higher than that of other lines. The spheroids of MARY-X completely disadhered when placed in media containing absent Ca(2+) or anti-E cadherin antibodies or when retrovirally transfected with a dominant-negative E cadherin mutant (H-2K(d)-E-cad). Anti-E-cadherin antibodies injected i.v. immunolocalized to the pulmonary lymphovascular emboli of MARY-X and caused their dissolution. H-2K(d)-E-cad-transfected MARY-X spheroids were only weakly tumorigenic and did not form lymphovascular emboli. A total of 90% of human IBCs showed increased membrane E-cadherin/alpha,beta-catenin immunoreactivity. These findings indicate that it is the gain and not the loss of the E-cadherin axis that contributes to the IBC phenotype. PMID- 11431365 TI - Entry routes of malignant lymphoma into the brain and eyes in a mouse model. AB - We have recently developed a novel mouse model for studying the infiltration of malignant lymphoma to the eye and brain. After i.p. inoculation of variant S49 mouse lymphoma cells into young mice (optimum: day 7 postnatal), specific homing of these cells (named Rev-2-T-6) to the brain and eyes took place. This model offers an opportunity to study the routes of infiltration to these sites and spread thereof, as well as the molecular mechanisms that govern this metastasis. By applying a time course histopathological analysis, we demonstrate that infiltration of the brain and eyes can be visualized as early as days 9 and 14 after inoculation, respectively. The lymphoma cells enter the brain preferentially through the choroid plexus and cranial nerves. Infiltration of the rostral part occurs before the caudal part of the brain. Once within the brain, the cells spread within it as well as migrate along the optic nerve sheath into the eyes, where they continue to migrate along the choroid, ciliary body, iris, and into the anterior chamber of the eye. The orbit is also infiltrated by the lymphoma cells. However, this occurs independent of the brain-optic nerve intraocular route. PMID- 11431366 TI - Influence of beta1 integrins on epidermal squamous cell carcinoma formation in a transgenic mouse model: alpha3beta1, but not alpha2beta1, suppresses malignant conversion. AB - Although aberrant integrin expression has been documented in many epithelial tumors, little is known about how integrins influence neoplastic progression. To examine this issue, transgenic mice in which the alpha2beta1 or alpha3beta1 integrin was expressed in the suprabasal epidermal layers via the involucrin promoter were subjected to skin carcinogenesis. Equal numbers of benign squamous papillomas were observed in transgenic and wild-type animals. However, the frequency of conversion of papillomas to malignant squamous cell carcinomas was much lower in alpha3beta1 transgenic than in alpha2beta1 transgenic and wild-type mice. No differences were observed in apoptosis or in the expression of endogenous integrins in transgenic and wild-type papillomas. However, alpha3beta1 transgenic papillomas displayed a diminished proliferative capacity and were more highly differentiated as judged by BrdUrd incorporation and keratin 10 expression, respectively, than alpha2beta1 transgenic and wild-type papillomas. Two proteins that associate with alpha3beta1 and not alpha2beta1 are extracellular matrix metalloproteinase inducer and CD81. Extracellular matrix metalloproteinase inducer expression correlated inversely with the degree of differentiation in normal epidermis and in transgenic and wild-type papillomas. Up-regulation of CD81 was observed in 100% of wild-type and 88% of alpha2beta1 transgenic papillomas but in only 25% of alpha3beta1 transgenic papillomas. CD81 was undetectable in untreated epidermis and strongly expressed in all transgenic and wild-type squamous cell carcinomas. Our results demonstrate that the alpha3beta1 integrin can suppress malignant conversion, and that the mechanism may involve CD81. PMID- 11431367 TI - Reduced blood vessel formation and tumor growth in alpha5-integrin-negative teratocarcinomas and embryoid bodies. AB - Embryonic stem (ES) cells-wild-type, heterozygous, or null for alpha5-integrin were injected ectopically into syngeneic mice to develop teratocarcinomas. alpha5 null-derived teratocarcinomas were significantly smaller than the wild-type or alpha5 heterozygous tumors. Histological analysis revealed the presence of tissues derived from all three germ layers, in all tumors. However, alpha5-null teratocarcinomas displayed less undifferentiated tissue than did the controls. Decreased proliferation and increased apoptosis were observed in the undifferentiated areas of the alpha5-null teratocarcinomas. The expression of extracellular matrix proteins, fibronectin and tenascin-C, and the basement membrane components, laminin, entactin/nidogen, and collagen IV, was similar in the different tumors, although the deposition of these molecules was more disorganized in alpha5-null teratocarcinomas. The absence of alpha5-integrin in the various tissues of the alpha5-null tumors was confirmed by immunohistochemistry. Many vessels, but not all, stained positively for alpha5 integrin, showing that they were host derived. Analysis of the area occupied by vessels revealed, on average, an 8-fold decrease in alpha5-null teratocarcinomas compared with control tumors. Staining for smooth muscle alpha-actin showed that pericytes and smooth muscle cells were recruited around the vessels in all tumors, suggesting similar vessel differentiation. Deposition of EIIIA and EIIIB and fibronectin around the vessels was observed in all tumors. The fact that some, although few, alpha5-integrin-negative vessels existed in alpha5-null tumors indicated that alpha5-/- ES cells could differentiate into endothelial cells. Endothelial cell differentiation and vessel formation were analyzed also in vitro. alpha5-null ES cells were differentiated into embryoid bodies, although they were delayed in growth and attachment. Differentiation into endothelial cells was achieved, but the organization into a complex vasculature was delayed compared with controls. We conclude that alpha5beta1-integrin plays a significant role in vessel formation both in ES cell cultures and in teratocarcinomas. Reduced vascularization likely contributed to the reduced proliferation and increased apoptosis observed in alpha5-null teratocarcinomas. PMID- 11431368 TI - Carbonic anhydrase IX, an endogenous hypoxia marker, expression in head and neck squamous cell carcinoma and its relationship to hypoxia, necrosis, and microvessel density. AB - Carbonic anhydrase IX (CA IX) is a transmembrane glycoprotein with an active extracellular enzyme site. We have shown previously that it was hypoxia inducible and may therefore be an endogenous marker of hypoxia. It is overexpressed in some tumors, particularly renal cell carcinoma. The aim of this study was to examine the expression and localization of CA IX in head and neck squamous cell carcinoma (HNSCC) and relate this to the location of tumor microvessels, angiogenesis, necrosis, and stage. Expression of CA IX was determined by immunoblotting in three HNSCC cell lines grown in normoxia and hypoxia (pO(2) 0.1%) and three paired tumor and normal tissue samples of HNSCC. Archived paraffin sections (79) of HNSCC were immunostained with antibodies to CA IX and CD34 to determine microvessel density (MVD). By double staining sections with CA IX and CD34, the distance between blood vessels and the start of CA IX expression and necrosis was calculated. CA IX was induced by hypoxia in all three HNSCC cell lines and overexpressed in HNSCC tumor tissue. Overexpression was localized to the perinecrotic area of the tumor on immunostaining, and the percentage area of the tumor expressing CA IX was significantly higher with more tumor necrosis (P = 0.001), a high MVD (P = 0.02), and advanced stage (P = 0.033) on univariate analysis and necrosis (P = 0.0003) and MVD (P = 0.0019) on multivariate analysis. The median distance between a blood vessel and the start of CA IX expression was 80 microm (range, 40-140 microm). CA IX is overexpressed in HNSCC because of hypoxia and is a potential biomarker for hypoxia in this tumor. Overexpression may help to maintain the intracellular pH, giving tumor cells a survival advantage and enhancing resistance to radiotherapy and chemotherapy. CA IX is a potential target for future therapy in HNSCC. PMID- 11431369 TI - Clonal variation in the B-lineage acute lymphoblastic leukemia response to multiple cytokines and bone marrow stromal cells. AB - The acquisition of genetic abnormalities in human B-lineage acute lymphoblastic leukemia (ALL) culminates in the clonal expansion of bone marrow (BM)-derived leukemic blasts. However, the response of leukemic cells to signals transduced by the BM microenvironment is not completely understood. The present study describes a new human B-lineage ALL cell line designated BLIN-4 (B LINeage-4). BLIN-4 cells respond to multiple cytokines/human BM stromal cell-derived molecules. One subline (BLIN-4E) undergoes cell death in the absence of BM stromal cells or cytokines and slowly proliferates on human BM stromal cells supplemented with interleukin (IL)-7 + FLT3-ligand. Another subline (BLIN-4L) slowly proliferates in the absence of cytokines and BM stromal cells and shows robust proliferation on BM stromal cells supplemented with IL-7 + FLT3-ligand. Although human BM stromal cells are comparable with IL-7 + FLT3-ligand in supporting proliferation of BLIN-4L cells, neutralizing antibody experiments demonstrate that BLIN-4L expansion on BM stromal cells is IL-7/FLT3-ligand independent. BLIN-4L could also respond to human thymic stromal lymphopoietin. BLIN-4E and BLIN-4L have the identical immunoglobulin heavy chain rearrangement and a CD10(+)/CD19(+)/CD20( )/CD22(+)/CD40(+)/mu heavy chain(-) phenotype. The original BM leukemic blasts harbored a ring chromosome 4 with a low percentage of cells also having either trisomy 8 or trisomy 18. The BLIN-4 sublines maintained the ring chromosome 4, but the trisomy 8 and trisomy 18 segregated into BLIN-4E and BLIN-4L, respectively. Thus, the BLIN-4 sublines exhibit biological characteristics consistent with a potential evolution in B-lineage ALL involving subclones with decreasing requirements on the BM microenvironment. PMID- 11431370 TI - A CD44 survival pathway triggers chemoresistance via lyn kinase and phosphoinositide 3-kinase/Akt in colon carcinoma cells. AB - A major obstacle to successful treatment of colorectal cancer is chemotherapy resistance. Enhanced expression of variant CD44 isoforms has been associated with aggressive tumor behavior, prompting the question of whether signaling from this receptor might modulate drug sensitivity. Activation of variant CD44 in colon carcinoma cell lines triggered resistance to the drug 1,3-bis(2-chloroethyl)-1 nitrosurea. Resistance was induced by monoclonal antibodies directed against epitopes independent of the hyaluronate-binding region but was not triggered by identical treatment of a carcinoma line expressing the standard CD44 isoform. We observed that variant CD44 produced activation of the src-family tyrosine kinase lyn. Moreover, overexpression of dominant-active lyn recapitulated chemoresistance via a pathway shown to involve activation of phosphoinositide 3 kinase and Akt. These results establish a novel role for CD44 in determining survival of colon carcinoma cells through lyn kinase and Akt. The ability to suppress apoptosis might play a critical role in the onset and development of colorectal malignancies. PMID- 11431371 TI - Overexpression of KAI1 suppresses in vitro invasiveness and in vivo metastasis in breast cancer cells. AB - KAI1 is a metastasis suppressor gene for human prostate cancer and is also involved in the progression of a variety of other human cancers. Previously, we have demonstrated that KAI1 expression was down-regulated in metastatic breast cancer cell lines as well as in highly aggressive breast cancer specimens. To determine whether KAI1 expression is responsible for the metastasis suppression in breast cancer, we transfected the human KAI1 cDNA into two highly malignant breast cancer cell lines, LCC6 and MDA-MB-231, which both have low levels of endogenous KAI1 expression. Parental, vector-only transfectants and KAI1 transfectant clones were injected into the mammary fat pads and tail veins, respectively, of athymic nude mice and assessed for both spontaneous and experimental lung metastasis. High KAI1 expression significantly suppressed the metastatic potential of KAI1-transfected LCC6 cells. Metastasis suppression correlated with the reduced rate of tumor growth and a decreased clonogenicity in soft agar. Furthermore, KAI1 expression significantly suppressed the in vitro cell invasion in KAI1-transfected MDA-MB-231 cells. Our results suggested that KAI1 may function as a negative regulator of breast cancer metastasis. PMID- 11431372 TI - Role of iron in tumor cell protection from the pro-apoptotic effect of nitric oxide. AB - F2The host defense against tumor cells is in part based upon the production of nitric oxide (NO) by activated macrophages. However, carcinogenesis may involve mechanisms that protect tumor cells from NO-mediated apoptosis. In the present study, we have assessed the effects of exogenous NO on the proliferation and survival of human liver (AKN-1), lung (A549), skin (HaCat), and pancreatic (Capan 2) tumor cell lines, compared with normal skin-derived epithelial cell cultures. Except to the HaCat cell line, all of the other human epithelioid cells were sensitive to the antiproliferation effect of S-nitroso-N-acetyl-penicillamine or Deta NONOate, whereas tumor cells had low if any response to sodium nitroprusside. Growth inhibition with exogenous NO correlated with increased apoptosis, but was not mediated by cyclic GMP, peroxynitrite generation, or poly(ADP-ribose) polymerase modulation, all of which involved in NO-mediated growth inhibition of normal skin-derived epithelial cell cultures. The simultaneous addition of iron-containing compounds protected tumor cells from NO mediated growth inhibition and apoptosis. Intracellular iron quantification indicated that, as deferoxamine, exogenous NO significantly decreased intracellular ferric iron levels in tumor cells. Together, the current study reveals that intracellular iron elevation rescues tumor cells from NO-mediated iron depletion and subsequent growth inhibition and apoptosis. PMID- 11431373 TI - A rare premalignant prostate tumor epithelial cell syndecan-1 forms a fibroblast growth factor-binding complex with progression-promoting ectopic fibroblast growth factor receptor 1. AB - The abnormal appearance and age-dependent loss of resident fibroblast growth factor receptor-2 (FGFR2) and gain of activity of FGFR1 in epithelial cells is a hallmark of the slow progression to malignancy in some models of prostate cancer. Pericellular matrix heparan sulfate (HS) is an integral subunit of the FGFR tyrosine kinase complex that restricts activity in absence of FGF, facilitates binding of an activating FGF, and confers specificity for FGF isoforms. In this report, we isolated and purified HS proteoglycan (HSPG) from premalignant prostate tumor epithelial cells based on the ability of the HS chains to form a binary complex with immunoglobulin module II of the ectopic and progression promoting FGFR1 that was competent to bind FGF. The FGFR1 affinity-purified product exhibited a specific activity of over 600 times that of crude cellular HSPG enriched from cell lysates by ion exchange chromatography. The purified preparation exhibited a single NH(2)-terminal sequence with 11 of 13 residues identical to syndecan-1. The activity of purified recombinant glutathione S transferase-tagged syndecan-1 expressed in premalignant epithelial cells confirmed that syndecan-1 bears HS chains that exhibit the rare motif that forms the FGF-binding complex with ectopic FGFR1. These results are the first to identify by affinity purification a specific HSPG core protein, the HS chains of which act as an integral subunit of the FGFR complex. The results suggest that syndecan-1 provides HS chains in premalignant epithelial cells to both the FGFR2- and FGFR1-signaling complexes that are integral to their dual roles in progression to malignancy. PMID- 11431374 TI - Promoter hypermethylation: a common cause of reduced p16(INK4a) expression in uveal melanoma. AB - Tumors often display unrestricted cell cycling attributable to a dysfunctional G(1)-S checkpoint. One of the mechanisms leading to such a defect is the inactivation of the cyclin-dependent kinase inhibitor p16(INK4a). Although inactivation of p16(INK4a) is observed in a wide range of tumors, including cutaneous melanoma, genetic alteration of p16(INK4a) is reportedly uncommon in uveal melanoma. Here we show that the p16(INK4a) promoter is hypermethylated in 6 of 12 uveal melanoma cell lines and in 7 of 22 primary uveal melanomas analyzed. Five of seven patients with a methylated primary tumor died of metastatic disease compared with 2 of 15 patients with a nonmethylated primary tumor. We also show that all uveal melanoma cell lines with a hypermethylated p16(INK4a) promoter have lost p16(INK4a) expression but have maintained the expression of p14(ARF). Treatment of uveal melanoma cell lines with 5-aza-2'-deoxycytidine results in demethylation of p16(INK4a) and in reexpression of p16(INK4a) mRNA, which is maintained upon withdrawal of the 5-aza-2'-deoxycytidine. In conclusion, p16(INK4a) promoter methylation appears to be a common event in uveal melanoma and is accompanied by the loss of p16(INK4a) expression. PMID- 11431375 TI - Mutational analysis of the peroxisome proliferator-activated receptor gamma gene in human malignancies. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) plays an important role in adipocyte differentiation and is expressed in many human malignancies, including those from prostate, breast, as well as colon. It regulates differentiation and/or cell growth of these cells. However, expression of this nuclear hormone receptor in other types of cancer, especially in hematological malignancies, remains to be fully elucidated. The PPARgamma gene has been mapped to chromosome band 3p25, where chromosomal abnormalities are observed in a variety of human malignancies. Furthermore, a recent study revealed that the PPARgamma gene is functionally mutated in sporadic colon cancer cells. Therefore, PPARgamma could be an important tumor suppressor gene. This prompted us to investigate the expression and mutational status of the PPARgamma gene in cancers of a variety of tissues. A total of 159 samples were interrogated for their expression of PPARgamma as measured by reverse transcription-polymerase chain reaction and/or Western blot analysis. In each of the samples, expression of PPARgamma was detectable. In addition, a total of 397 clinical samples and cell lines including colon, prostate, breast and lung cancers, and leukemias were analyzed for mutations of the PPARgamma gene by either reverse transcription polymerase chain reaction-single-strand conformation polymorphism or polymerase chain reaction-single-strand conformation polymorphism analysis. No abnormalities were detectable in any of the human malignancies. On the other hand, shifted bands were easily detectable when using positive controls, which harbored the same sequence alterations reported previously in colon cancer cells. Taken together, PPARgamma is expressed in a variety of cancers, and mutation of the PPARgamma gene is a very rare event in human malignancies. PMID- 11431376 TI - The COOH-terminal domain of FLI-1 is necessary for full tumorigenesis and transcriptional modulation by EWS/FLI-1. AB - More than 85% of Ewing's family tumors carry a specific chromosomal translocation that fuses the NH(2) terminus of the EWS gene to the COOH terminus of the FLI1 transcription factor. It has been shown previously that both the transactivation domain encoded by EWS and the DNA binding domain of FLI1 were necessary for transforming cells to anchorage independence. We now report that a COOH-terminal domain in addition to the FLI1 DNA binding domain is necessary to promote cellular transformation. NIH 3T3 cells expressing a COOH-terminal deletion mutant (EWS/FLI1 DeltaC) have a greatly reduced capability to form colonies in soft agar and tumors in severe combined immunodeficient mice. The rate of tumor formation for NIH 3T3 that express EWS/FLI1 DeltaC is 50 days, whereas EWS/FLI1 forms tumors within 22 days. In addition, cells expressing the EWS/FLI1 DeltaC mutant failed to completely demonstrate the round-cell histology that is seen in both Ewing's tumor cell lines and NIH 3T3 cells expressing full-length EWS/FLI1. Northern and microarray analyses were performed to assess the effect of loss of the FLI1 COOH terminus on transcriptional modulation of EWS/FLI1 target genes. We found that although EWS/FLI1 DeltaC up-regulates smaller numbers of genes (21 genes) compared with EWS/FLI1 (34 genes), 41% of the EWS/FLI1 targets were also up-regulated by EWS/FLI1 DeltaC. On the other hand, EWS/FLI1 DeltaC is unable to down-regulate genes (3 genes) as efficiently as EWS/FLI1 (39 genes) with only one target gene repressed by both fusion constructs. Our study indicates that the EWS/FLI1 transcription factor has strong transcriptional activating as well as repressing properties and suggests that transcriptional activation and repression of target genes may occur through biochemically different mechanisms. PMID- 11431377 TI - Effect of changing tumor oxygenation on glycolytic metabolism in a murine C3H mammary carcinoma assessed by in vivo nuclear magnetic resonance spectroscopy. AB - The rate of conversion of D-[1-(13)C]glucose into [3-(13)C]lactate (apparent glycolytic rate) has been determined in C3H murine mammary carcinomas in vivo using tumor-selective (13)C nuclear magnetic resonance spectroscopy with (1)H (13)C cross-polarization. Under conditions of acute hypoxia induced by breathing carbon monoxide at 660 ppm, the apparent glycolytic rate was 0.0239 +/- 0.0019 min(-1). The proportion of (13)C label incorporated into [4-(13)C]glutamate (measured in tumor extracts) was 25-fold lower than that incorporated into [3 (13)C]lactate, reflecting a very limited oxidative metabolism during this hypoxic episode. For animals breathing air or carbogen (95% O(2) + 5% CO(2)), the calculated glycolytic rates were correspondingly lower (0.0160 +/- 0.0021 min(-1) and 0.0050 +/- 0.0011 min(-1), respectively). Although (13)C labeling of glutamate at C4 was still an order of magnitude lower than that for lactate at C3 (11-fold for air and 9-fold for carbogen), these ratios did show a greater degree of oxidative metabolism than that seen in animals breathing carbon monoxide at 660 ppm. The marked difference in apparent glycolytic rate for this tumor model between well-oxygenated and hypoxic conditions demonstrates a substantial Pasteur effect (inhibition of glycolysis by oxygen). Dynamic (13)C nuclear magnetic resonance spectroscopy provides a noninvasive estimate of tumor glycolysis that can be used to evaluate the relationship between oxygenation and energy metabolism, and this has potential consequences for the sensitivity of hypoxic cells to treatment and their ability to promote angiogenesis. PMID- 11431380 TI - Does continuity of care matter? Yes: consistent contact with a physician improves outcomes. PMID- 11431381 TI - Does continuity of care matter? No: discontinuity can improve patient care. PMID- 11431382 TI - Why should physicians be concerned about health inequalities? Because inequalities are unfair and hurt everyone. PMID- 11431386 TI - FDA warning to manufacturers of AIDS drugs. PMID- 11431385 TI - Three papules in a row. PMID- 11431387 TI - Breast cancer patients can stop tamoxifen after 5 years. PMID- 11431389 TI - Metaphorically speaking: the metaphor of health care provision as a factory. PMID- 11431390 TI - Abdominal injuries from snowboarding. PMID- 11431391 TI - Meaning, soul, state of mind, and hope. PMID- 11431393 TI - More on living wills, death, and dying. PMID- 11431394 TI - Effects of being uninsured on ethnic minorities' management of chronic illness. AB - OBJECTIVE: To compare the effectiveness with which insured and uninsured persons with chronic illnesses managed their health care. DESIGN: Recruited volunteers diagnosed with a variety of chronic illnesses who underwent 3 semistructured interviews in a 1-year period. SETTING: Volunteers were recruited through referrals, flyers, and face-to-face contacts from community health clinics, senior centers, acute care hospitals, and home care services in 2 urban counties in California between December 1997 and December 2000. PARTICIPANTS: A total of 297 persons between the ages of 23 and 97 years (35% African American, 33% Latino, and 32% Filipino American), of whom 42 (14%) had no health insurance. MAIN OUTCOME MEASURES: Qualitative analysis of interview data compared insured and uninsured respondents on a series of components of chronic illness management, including control over illness, frequency of health crises, procuring medication, use of medication, understanding of the illness, knowledge of self care measures, and awareness of risk factors. Whether respondents were under the care of a regular physician was also assessed. RESULTS: Compared with insured respondents, uninsured respondents were much less effective at managing their illnesses. The uninsured had poorly controlled illnesses, frequent health crises, difficulty procuring medication, used medication incorrectly, demonstrated poor understanding of their illness, and displayed little knowledge of self-care measures or risk awareness. They rarely had a regular physician or attended a specific health clinic. CONCLUSIONS: The findings suggest that not only did uninsured persons with chronic health conditions lack adequate health care, their illnesses were also poorly controlled. Inadequately educated about their health, uninsured persons lacked the information, insight, and tools that would have allowed them to manage their illnesses more effectively. PMID- 11431395 TI - Charting the path from lack of insurance to poor health outcomes. PMID- 11431398 TI - Waiting for conflict before requesting an ethics consultation. PMID- 11431399 TI - Menopause and hormone-replacement therapy: Part 2. Hormone-replacement therapy regimens. PMID- 11431400 TI - Treating the patient or the population? Part 1. Judging the benefit of treatment of individual patients. PMID- 11431401 TI - Assessing diagnostic and screening tests: Part 2. How to use the research literature on diagnosis. PMID- 11431403 TI - Genetics through a primary care lens. PMID- 11431405 TI - Art therapy with adolescents. PMID- 11431406 TI - Managing medicine: a new breed of medical businessperson is needed for the 21st century. PMID- 11431407 TI - Empathy: a lost meaning? PMID- 11431408 TI - The master's in public health: a better preparation for understanding the health care system. PMID- 11431409 TI - Miracles. PMID- 11431410 TI - Immigrant women's health: screening and immunization. PMID- 11431412 TI - The solitary vice: the superstition that masturbation could cause mental illness. PMID- 11431414 TI - Regulation of mouse mast cell surface Fc epsilon RI expression by dexamethasone. AB - It is now clear that the mast cell's functional response to IgE-dependent stimulation can be influenced significantly by the level of expression of the high-affinity IgE receptor (Fc epsilon RI) on the cell's surface. Thus, modulation of Fc epsilon RI surface expression represents a potentially important mechanism for regulating mast cell activity in allergic reactions. In this study, we examined whether a glucocorticoid, dexamethasone (DEX), can influence levels of mast cell Fc epsilon RI expression either in the presence or absence of IgE, an up-regulator of the mast cell surface Fc epsilon RI level. In the absence of IgE, DEX decreased the surface Fc epsilon RI levels in mouse peritoneal mast cells, mouse bone marrow-derived cultured mast cells and a mouse mast cell line, Cl.MC/C57.1. Moreover, DEX also partially suppressed the ability of IgE to enhance surface expression of Fc epsilon RI in these cells. Three different glucocorticoids, DEX, methylprednisolone and hydrocortisone, suppressed Fc epsilon RI expression in mast cells, whereas sex steroids, i.e. estradiol, progesterone and testosterone, did not, indicating that the Fc epsilon RI suppressing effect is glucocorticoid specific. On the other hand, DEX did not affect levels of Fc epsilon RI alpha, beta or gamma mRNA, suggesting that its ability to decrease surface Fc epsilon RI reflects a post-transcriptional mechanism. Finally, DEX-treated mast cells showed a reduced degranulation response to antigenic stimulation through down-regulation of surface Fc epsilon RI expression in addition to DEX-induced changes in downstream signals. These results show that mast cell surface Fc epsilon RI expression is suppressed by glucocorticoids in both the presence and absence of IgE, and suggest that reduction of mast cell surface Fc epsilon RI levels may be one of the favorable anti-allergic actions of glucocorticoids. PMID- 11431415 TI - Crucial amino acid residues of mouse CD1d for glycolipid ligand presentation to V(alpha)14 NKT cells. AB - A novel lymphocyte, NKT cells bearing an invariant V(alpha)14 antigen receptor, specifically recognizes alpha-galactosylceramide (alpha-GalCer) exclusively presented by mouse CD1d (mCD1d). However, the precise molecular interaction remains unclear. For the basis of functional analyses, a docking model of alpha GalCer with the crystal structure of mCD1d was constructed. Possible residues involved in the alpha-GalCer--mCD1d interaction were found to be Arg79, Glu83 and Asp80 for carbohydrate recognition, and Asp153 for interaction with the amide group on the fatty acyl chain. The alpha-GalCer-presenting ability of various transfectants expressing mutant mCD1d was completely abrogated if a single amino acid mutation was induced at positions 79, 80, 83 or 153, suggesting that the polar amino acids above the F' pocket are crucial for alpha-GalCer presentation to activate V(alpha)14 NKT cells. The possibility that Glu83 is a contact site for the NKT cell receptor is also discussed. PMID- 11431416 TI - The peptide-specific alloreactive human T cell repertoire varies largely between individuals and is not extended in HLA-A*0205--anti-HLA-A*0201 pairings. AB - Alloreactive T cells recognize framework or peptide-dependent determinants on foreign MHC molecules. Among the peptide-dependent alloreactive T cells a significant proportion is specific for one particular peptide presented by the allo-MHC molecule as antigen-specific T cells would do. Such alloreactive, peptide-specific T cells are referred to as 'allorestricted'. High-avidity HLA A*02 allorestricted cytotoxic T lymphocyte (CTL) clones specific for peptide libraries can be generated from HLA-A*02(-) donors. We made use of this technique to study the role of closely related self-HLA molecules on shaping of the alloreactive T cell repertoire. Peripheral blood lymphocytes from HLA-A*0205 individuals were stimulated by HLA-A*0201 targets pulsed with an HLA-A*0201 peptide library. We did not observe a bias towards peptide-specific CTL in the HLA-A*0201-directed alloreactive repertoire of HLA-A*0205 donors as compared to HLA-A*02(-) donors. Comparison of the alloreactive T cell response between two donors having similar HLA haplotypes demonstrated that the allorestricted T cell repertoire is largely different between individuals. PMID- 11431417 TI - B cell co-receptors regulating T cell-dependent antibody production in common variable immunodeficiency: CD27 pathway defects identify subsets of severely immuno-compromised patients. AB - CD27 and CD134 ligand (CD134L) are two B cell co-receptors for T(h) cell activation-induced ligands (i.e. CD70 and CD134) that promote differentiation of B cells into plasma cells and high-rate antibody production respectively. We explored the CD27 pathway and T cell CD134 expression in common variable immunodeficiency (CVID), a disease characterized by a lack of plasma cells and low Ig serum levels. Twelve patients were compared to seven healthy controls. We found a low percentage of circulating CD27(+) B cells in seven patients and B cell CD27 expression was not up-regulated by in vitro activation in two of them. Importantly, the number of circulating CD27(+) B cells was correlated with the severity of the disease--the patients with the lowest CD27(+) B cell counts having the lowest serum Ig concentrations and the lowest total peripheral blood B cell counts. In contrast, CD70 and CD134 were normally expressed on in vitro activated T cells. CD134L was not detected on patient and control B cells in our activation conditions. Functional studies of in vitro Ig production demonstrated an absence of B cell response to CD27 cross-linking, in particular in a patient with normal CD27 expression. Our results indicate that a defect in CD27 expression or function contributes to the pathogenesis of certain severe forms of CVID. PMID- 11431418 TI - Degeneracy and additional alloreactivity of drug-specific human alpha beta(+) T cell clones. AB - It has been well established that T cells can recognize small mol. wt compounds such as drugs. Results from previous studies revealing a high heterogeneity and cross-reactivity of drug-specific T cell clones (TCC) in individual patients prompted us to analyze the degeneracy of drug-reactive TCR in detail. Hence, we analyzed the MHC restriction pattern of a panel of 100 drug-specific TCC isolated from different drug-allergic donors. We found that 28 of the tested clones showed an MHC allele-unrestricted drug recognition. Most of these clones were at the same time highly drug specific, i.e. they could only be stimulated by the original drug and not by any drug derivatives. In contrast, TCC with the ability to interact with different drug derivatives displayed a clearly MHC allele restricted drug recognition. Therefore, we concluded that the TCR of these clones is mainly interacting with side chains of the appropriate drug molecules and hence able to tolerate alterations in the MHC molecule. Moreover, we tested all clones for additional alloreactivity and found that 27 clones could be stimulated by a self-MHC--peptide--drug complex as well as by a non-self-MHC--peptide complex. This cross-reactivity with allogeneic MHC molecules was substantially higher in drug-specific TCC compared to tetanus toxoid-specific clones from the same donors. This suggests that from the point of view of drug-specific TCR, non self-MHC--peptide complexes have a higher incidence to mimic the 'original' self MHC--peptide-drug complex and this may occur for TCR recognizing self-MHC- pathogen-derived peptide complexes. Finally, the biological functions of bispecific TCC were not influenced by the nature of the stimulating ligand. Both drug as well as allogeneic stimulation led to similar reaction patterns in the analyzed TCC. PMID- 11431419 TI - Cytometric and functional analyses of NK and NKT cell deficiencies in NOD mice. AB - Defects in NK and NKT cell activities have been implicated in the etiology of type 1 (autoimmune) diabetes in NOD mice on the basis of experiments performed using surrogate phenotypes for the identification of these lymphocyte subsets. Here, we have generated a congenic line of NOD mice (NOD.b-Nkrp1(b)) which express the allelic NK1.1 marker, enabling the direct study of NK and NKT cells in NOD mice. Major deficiencies in both populations were identified when NOD.b Nkrp1(b) mice were compared with C57BL/6 and BALB.B6-Cmv1(r) mice by flow cytometry. The decrease in numbers of peripheral NK cells was associated with an increase in their numbers in the bone marrow, suggesting that a defect in NK cell export may be involved. In contrast, the most severe deficiency of NKT cells found was in the thymus, indicating that defects in thymic production were probably responsible. The deficiencies in NK cell activity in NOD mice could only partly be accounted for by the reduced numbers of NK cells, and fewer NKT cells from NOD mice produced IL-4 following stimulation, suggesting that NK and NKT cells from NOD mice shared functional deficiencies in addition to their numerical deficiencies. Despite the relative lack of IL-4 production by NOD NKT cells, adoptive transfer of alpha beta TCR(+)NK1.1(+) syngeneic NKT cells into 3-week old NOD recipients successfully prevented the onset of spontaneous diabetes. As both NK and NKT cells play roles in regulating immune responses, we postulate that the synergistic defects reported here contribute to the susceptibility of NOD mice to autoimmune disease. PMID- 11431420 TI - Mechanisms of cytokine synergy essential for vaccine protection against viral challenge. AB - The ability of cytokines to steer CD4(+) T(h) cell responses toward a T(h)1 or T(h)2 phenotype and enhance the magnitude of both CD8(+) cytotoxic T lymphocytes (CTL) and antibody responses has clearly been demonstrated by our lab and others, but the influence of cytokines on protective immune responses is much less clear. Here we show an essential role for CD4(+) T(h)1 helper cell induction and IFN gamma production in protection from viral challenge with a recombinant vaccinia virus expressing HIV-1MN viral envelope glycoprotein gp160. Complete protection from viral challenge is achieved only when the triple combination of exogenous cytokines granulocyte macrophage colony stimulating factor (GM-CSF), IL-12 and tumor necrosis factor (TNF)-alpha are co-administered with the peptide vaccine. In vivo depletion of CD4(+) cells or immunization of IFN-gamma-deficient mice abrogates protection. GM-CSF, IL-12 and TNF-alpha also synergize for the enhanced induction of CTL; however, adoptive transfer of a CD8(+) CTL line afforded only partial protection in this viral challenge model. As a possible mechanism of in vivo protection we show that GM-CSF increases the percentage and activity of antigen-presenting dendritic cells in draining lymph nodes where the immune response is initiated. We further demonstrate synergy between IL-12 and the proinflammatory cytokine TNF-alpha in driving IFN-gamma production. Thus, a combination of IL-12 and TNF-alpha is essential for the optimal development of T(h)1 responses and help for CTL induction in BALB/c mice, and is complemented by a third cytokine, GM-CSF, which enhances antigen presentation. PMID- 11431421 TI - Identification of conformation-dependent epitopes and V gene selection in the B cell response to type II collagen in the DA rat. AB - Collagen-induced arthritis (CIA) is a widely used model for rheumatoid arthritis. Induction of CIA in rats using rat type II collagen (CII) results in a chronic arthritis in which anti-CII antibodies are believed to play a pathogenic role. In this study, we analyzed the epitope selection and V gene usage in the anti-CII response in the DA rat. A panel of CII-reactive B cell hybridomas was established from the draining lymph nodes 11 days after immunization. All of the CII-specific antibodies bound cartilage in vivo, showing that these are true autoantibodies. These antibodies were all IgG and specific for several distinct triple helical epitopes on CII. Interestingly, the major epitope, recognized by four different antibodies, was identical with the major B cell epitope in the mouse CII located at position 359--369 (denoted as C1(III)). The Q52 and PC7183 V(H) gene families encoded 12 out of 14 sequenced heavy chains. There was a relatively more heterogeneous usage of V(L) genes as the antibodies were encoded by four different V(kappa) families (V(kappa)1, V(kappa)2, V(kappa)12/13 and V(kappa)RF). As in the mouse, some of the V genes used showed germline characteristics. We conclude that the immune response in the rat shares epitope specificity and a constrained V gene repertoire with the mouse. However, the V genes used for recognition of the closely related collagen structures differed considerably between mouse and rat, indicating an influence of the species-specific variation in the V gene repertoire. PMID- 11431422 TI - Tetracyclines inhibit activated B cell function. AB - Tetracyclines have recently been shown to exert a number of pleiotropic anti inflammatory and immunomodulatory activities, independent of their antibiotic properties. These include the ability to inhibit metalloproteinases (MP), a class of enzymes involved in crucial cellular functions such as the shedding of soluble mediators and their receptors from the cell surface, as well as interaction with, and remodeling of, the extracellular matrix. Here we report that doxycycline at therapeutic concentrations (1--5 microg/ml) significantly suppresses Ig secretion and class switching by in vitro activated murine B cells. Suppression of Ig secretion correlates with a decrease in levels of mRNA for the terminal B cell differentiation-associated genes Blimp-1 and mad-4, as well as to a reduction in expression of the plasma cell markers Syndecan-1 and J chain. Inhibition of class switching occurs at the recombination stage and is also induced by other MP inhibitors, including tetracycline analogs lacking antibiotic activity and the chemically unrelated hydroxamate KB8301. These novel, direct effects of MP inhibitors on B lymphocytes suggest an intrinsic role for MP in B cell activation and likely explain some of the observed in vivo immunomodulatory properties of tetracyclines. Moreover, these findings have significant implications for tetracycline therapy in Ig-mediated autoimmune or allergic diseases and raise questions about the use of doxycycline-inducible transgenic systems for the study of B cell function. PMID- 11431423 TI - Discrimination of bacterial lipoproteins by Toll-like receptor 6. AB - Bacterial lipoproteins (BLP) trigger immune responses via Toll-like receptor 2 (TLR2) and their immunostimulatory properties are attributed to the presence of a lipoylated N-terminus. Most BLP are triacylated at the N-terminus cysteine residue, but mycoplasmal macrophage-activating lipopeptide-2 kD (MALP-2) is only diacylated. Here we show that TLR6-deficient (TLR6(-/-)) cells are unresponsive to MALP-2 but retain their normal responses to lipopeptides of other bacterial origins. Reconstitution experiments in TLR2(-/-) TLR6(-/-) embryonic fibroblasts reveal that co-expression of TLR2 and TLR6 is absolutely required for MALP-2 responsiveness. Taken together, these results show that TLR6 recognizes MALP-2 cooperatively with TLR2, and appears to discriminate between the N-terminal lipoylated structures of MALP-2 and lipopeptides derived from other bacteria. PMID- 11431424 TI - The flexibility of the TCR allows recognition of a large set of naturally occurring epitope variants by HIV-specific cytotoxic T lymphocytes. AB - Pathogens attempt to evade immune recognition by expressing mutated antigens. The present study shows that two mechanisms happen in vivo during the course of HIV infection to limit the escape of antigenic variants from cytotoxic T lymphocyte (CTL) recognition: recognition of several epitope variants by the same TCR and generation of several CTL populations specific for a single epitope but recognizing different variant sequences. We have studied two CTL populations directed towards the HIV-p24gag amino acids 176--184 QASQEVKNW epitope, presented by HLA-B5301. Both CTL populations were derived from a long-term asymptomatic HIV infected child and they express different TCR. Each of the two CTL recognizes five of the 10 naturally occurring variants. These variants are distinct for both CTL and thus a total of eight variants are recognized. Thus, polyclonality of CTL specific for the same epitope but differing in variant sequences recognized may improve the control of variant viruses' replication in vivo. In addition to cross recognition of several variant epitopes, promiscuous recognition of exogenous peptides complexed to allogeneic HLA-B molecules occurs, showing that the TCR can tolerate amino acid changes on both the peptide and the MHC molecule. This flexibility of the TCR is probably of great importance for control of viruses with high genetic variability, such as HIV. PMID- 11431425 TI - Combinations of dominant-negative class II transactivator, p300 or CDK9 proteins block the expression of MHC II genes. AB - The class II transactivator (CIITA) regulates not only the transcription of HLA DR, -DQ, -DP, but also invariant chain, DMA and DMB genes. A hybrid mutant CIITA protein, which contained residues from positions 302 to 1130 in CIITA fused to the enhanced green fluorescent protein (EdCIITA), inhibited the function of the wild-type protein. EdCIITA extinguished the inducible and constitutive expression of MHC II genes in epithelial cells treated with IFN-gamma and B lymphoblastoid cells respectively. Also, it blocked T cell activation by superantigen. This inhibition correlated with the localization of EdCIITA but not CIITA in the cytoplasm of cells. However, when EdCIITA was co-expressed with a dominant negative form of the nucleoporin Nup214/CAN, it also accumulated in the nucleus. These data suggest that EdCIITA not only competes with the wild-type protein for the binding to MHC II promoters but sequesters a critical co-factor of CIITA in the cytoplasm. CIITA also recruits the histone acetyltransferase cAMP responsive element binding protein (CREB) binding protein and positive transcription elongation factor b (p-TEFb) for the transcription of MHC II genes. Dominant negative p300 (DNp300) or CDK9 (DNCDK9) proteins inhibited the function of CIITA and of the DRA promoter. Thus, combinations of EdCIITA and DNp300 and/or DNCDK9 proteins extinguished the transcription of MHC II genes. They might become useful for future genetic therapeutic approaches in organ transplantation and autoimmune diseases. PMID- 11431426 TI - A soluble form of CD83 is released from activated dendritic cells and B lymphocytes, and is detectable in normal human sera. AB - CD83 is an inducible glycoprotein expressed predominantly by dendritic cells (DC) and B lymphocytes. Expression of membrane CD83 (mCD83) is widely used as a marker of differentiated/activated DC but its function and ligand(s) are presently unknown. We report the existence of a soluble form of CD83 (sCD83). Using both a sCD83-specific ELISA and Western blotting, we could demonstrate the release of sCD83 by mCD83(+) B cell and Hodgkin's disease-derived cell lines, but not mCD83( ) cells. Inhibition of de novo protein synthesis did not affect the release of sCD83 during short-term (2 h) culture of cell lines although mCD83 expression was significantly reduced, suggesting sCD83 is generated by the release of mCD83. Isolated tonsillar B lymphocytes and monocyte-derived DC, which are mCD83(low), released only low levels of sCD83 during culture. However, the differentiation/activation of these populations both up-regulated mCD83 and increased sCD83 release significantly. Analysis of sera from normal donors demonstrated the presence of low levels (121 +/- 3.6 pg/ml) of circulating sCD83. Further studies utilizing purified sCD83 and the analysis of sCD83 levels in disease may provide clues to the function and ligand(s) of CD83. PMID- 11431427 TI - The genetics of cataract. PMID- 11431428 TI - Detection of c-myc amplification in uveal melanoma by fluorescent in situ hybridization. AB - PURPOSE: Genetic abnormalities of chromosomal arm 8q have been reported by many studies in uveal melanoma. To better understand the role of 8q abnormalities in uveal melanoma development, copy number anomalies of the c-myc oncogene (located on 8q24.1) have been investigated. METHODS: Forty-three uveal melanomas were analyzed by fluorescent in situ hybridization (FISH) with probes for c-myc and the chromosome 8 centromere. Results of the FISH analysis were compared with genetic changes previously detected by microsatellite analysis on chromosomes 3 and 6p. RESULTS: Thirty uveal melanomas (70%) had extra copies of c-myc, 2 tumors (5%) had loss of c-myc, and 11 tumors (25%) had no abnormalities in c-myc copy number. Of those with extra copies of c-myc, 13 tumors (43%) had amplification of the c-myc gene, 14 tumors (47%) had an intermediate relative increase in the c myc copy number, and 3 tumors (10%) had a simple gain of chromosome 8. An association between larger tumor size and c-myc amplification was found (P < 0.01). Although extra copies of c-myc were seen in tumors with retention of chromosome 3, remarkably only tumors with monosomy 3 showed amplification of c myc (P = 0.03). CONCLUSIONS: The specific amplification of the c-myc oncogene detected in at least 30% of primary uveal melanomas cannot be explained by the simple 8q abnormalities observed in cytogenetic studies. The striking association between c-myc amplification and monosomy 3 suggests a unique pathway of genetic progression in a subset of uveal melanomas. PMID- 11431429 TI - Delayed dark-adaptation and lipofuscin accumulation in abcr+/- mice: implications for involvement of ABCR in age-related macular degeneration. AB - PURPOSE: To examine the ocular phenotype in mice heterozygous for a null mutation in the abcr gene. METHODS: Retinas and retinal pigment epithelia (RPE) were prepared from wild-type, abcr+/-, and abcr-/- mice. Fresh tissues were homogenized and analyzed by normal phase high-performance liquid chromatography (HPLC) for the presence of retinoids and phospholipids. In another study, fixed tissues were sectioned and analyzed by light and electron microscopy. Finally, anesthetized mice were studied by electroretinography (ERG) at different times after exposure to strong light. RESULTS: A2E, the major fluorophore of lipofuscin, and its precursors, A2PE-H(2) and A2PE, were approximately fourfold more abundant in 8-month-old abcr+/- than in the wild-type retina and RPE. The levels of these substances in abcr+/- mice were approximately 40% those in abcr-/ mice. Lipofuscin pigment-granules were also visible in abcr+/- RPE cells by electron microscopy. Accumulation of A2PE-H(2) and A2E in abcr+/- retina and RPE, respectively, was strongly dependent on light exposure. Heterozygous mutants also exhibited delayed recovery of rod sensitivity by ERG. This delay was correlated with elevated levels of all-trans-retinaldehyde (all-trans-RAL) in retina after a photobleach and was not caused by a reduction in quantum-catch due to depletion of 11-cis-retinaldehyde (11-cis-RAL). CONCLUSIONS: Partial loss of the ABCR or rim protein is sufficient to cause a phenotype in mice similar to recessive Stargardt's disease (STGD) and age-related macular degeneration (AMD) in humans. These data are consistent with the suggestion that the STGD carrier-state may predispose to the development of AMD. PMID- 11431430 TI - The effect of TGF-beta1 on differential gene expression profiles in human corneal epithelium studied by cDNA expression array. AB - PURPOSE: TGF-betas regulate cell proliferation and differentiation, and they play important roles in maintenance of corneal epithelium. However, the precise function of TGF-betas in the corneal epithelium remains unclear. In this study, cDNA expression array technology was used to demonstrate the effect of TGF-beta1 on the simultaneous expression of a large number of genes in cultured human corneal epithelial cells (HCECs). The change in protein level expression of the specific genes influenced by TGF-beta1 was also investigated. METHODS: Human cDNA expression array technology was used to study the simultaneous expression of 1176 specific cellular genes in HCECs incubated with TGF-beta1 (10 ng/ml). Moreover, gene-specific semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to confirm the gene expression pattern measured by the cDNA expression array. Western blot analysis was used to examine protein expression of the specific genes in the presence or absence of TGF-beta1. RESULTS: TGF-beta1 significantly upregulated the expression of 19 genes and significantly downregulated ras-related protein, caspase10, and beta4-integrin in the treated HCECs. The expression of 277 genes including alpha3-integrin, PAI-2, transferrin receptor, and cyclin-D1 was studied. Semiquantitative RT-PCR analysis confirmed the TGF-beta1-mediated changes in expression patterns of these genes. Furthermore, Western blot analysis revealed that TGF-beta1 remarkably decreased PAI-2, transferrin receptor, and integrin alpha3, and increased caspase10 on the protein level. CONCLUSIONS: TGF-beta1 regulates the expression of specific types of genes in HCECs. These results strongly suggest that TGF-beta1 is critically involved in the maintenance of the corneal epithelium through the control of a network of various signal-transduction pathways. PMID- 11431431 TI - Induction of the ubiquitin-proteasome pathway during the keratocyte transition to the repair fibroblast phenotype. AB - PURPOSE: To examine dynamics and function of the ubiquitin (Ub)-proteasome pathway (UPP) during corneal stromal cell acquisition of the repair fibroblast phenotype. METHODS: An established cell culture model was used in which freshly isolated rabbit corneal stromal cells acquire a repair fibroblast phenotype, thereby mimicking injury-induced stromal cell activation. RESULTS: Transition to the repair fibroblast phenotype during the 72 hours after initial plating was coincident with progressive UPP induction. Levels of Ub, Ub-conjugated proteins, ubiquitinylating enzymes E1 and E2-25K, and 26 S proteasome increased two- to fivefold in activated stromal cells. These increases were associated with enhanced (>10-fold) capacity for Ub-dependent proteolysis of (125)I-labeled H2A and with progressive (>6-fold) increases in the UPP substrate, inhibitor of kappaBalpha (IkappaBalpha). Because IkappaBalpha expression is induced by nuclear factor (NF)-kappaB, this finding suggests that rates of constitutive NF-kappaB activation, and thus IkappaBalpha degradation, are elevated in activated stromal cells. Both freshly isolated and activated stromal cells degraded IkappaBalpha in response to IL-1alpha; yet, only activated stromal cells maintained autocrine IL 1alpha expression after 24 hours. UPP induction was coincident with a more than 90% loss of tissue transketolase (TKT) and aldehyde dehydrogenase (ALDH) class 1. TKT was stabilized during the repair phenotype transition by proteasome inhibition and was degraded (>30%/h) by the UPP in cell-free assays. CONCLUSIONS: Coordinate induction of the UPP during stromal cell activation alters levels of IkappaBalpha and TKT, two UPP substrates that are implicated in the loss of tissue stasis and corneal clarity after injury. PMID- 11431432 TI - The metabolism of fatty acids in human Bietti crystalline dystrophy. AB - PURPOSE: To investigate the role of abnormal lipid metabolism in Bietti crystalline dystrophy. METHODS: Cultured human lymphocytes and fibroblasts from patients with Bietti crystalline dystrophy (BCD) were incubated in the presence of [(14)C]18:3n-3 or [(14)C]18:2n-6. Incorporation into the cellular lipid pools and further metabolism by desaturation or elongation were monitored by thin-layer chromatography and HPLC. Results were compared with those in normal control subjects and patients with Wolman disease (WD). RESULTS: Pulse-chase experiments with labeled fatty acids in all groups showed that, after 1 hour, radioactivity was largely confined to the triacylglyceride (TG) and choline phosphoglyceride (CPG) pools. However, after several hours, radioactivity was transferred from the TG and CPG pools, some going to the serine and ethanolamine phosphoglyceride (SPG and EPG) pools. Fibroblasts from all groups showed direct transfer of fatty acids (FAs) into CPG and EPG. Incorporation of labeled FAs into the EPG pool paralleled extensive desaturation and elongation of 18:2n-6 to 22:5n-6 and 18:3n-3 to 22:6n 3. Fibroblasts from patients with WD (a lysosomal acid lipase deficiency characterized by excessive lipid accumulation), showed higher incorporation of 18:2n-6 into TGs than did normal or BCD fibroblasts. Conversely, fibroblasts from patients with BCD showed lower conversion of 18:3n-3, but not of 18:2n-6, into polyunsaturated FAs (PUFAs) than those of normal subjects or patients with WD. This was true for total FAs, CPGs, and EPGs. Similar results were found in both fibroblasts and lymphocytes; however, unlike fibroblasts, lymphocytes from normal subjects showed similar levels of incorporation of FAs into EPGs and CPGs. In contrast, incorporation of 18:3n-3 into EPGs was decreased in lymphocytes from patients with BCD. CONCLUSIONS: BCD is characterized by a lower than normal conversion of FA precursors into n-3 PUFA, whereas there is a higher than normal level of n-6 and n-3 FAs incorporation into TGs in cells from patients with WD. These findings raise the possibility that abnormal lipid metabolism associated with BCD is the result of deficient lipid binding, elongation, or desaturation in contrast to the lysosomal acid lipase deficiency found in Wolman disease. PMID- 11431433 TI - Low docosahexaenoic acid levels in rod outer segment membranes of mice with rds/peripherin and P216L peripherin mutations. AB - PURPOSE: Humans with retinitis pigmentosa and dogs with progressive rod-cone degeneration (prcd) have lower than normal blood levels of long-chain polyunsaturated fatty acids, including docosahexaenoic acid (DHA), the major fatty acid found in retinal rod outer segments (ROS). In addition, prcd-affected dogs have lower levels of DHA in their ROS than control animals. The present study was designed to determine whether mice that are heterozygous for the rds mutation and transgenic mice heterozygous for a specific rds/peripherin mutation (P216L) have lower DHA levels in their ROS and other tissues than do control mice. METHODS: Wild-type (rds(+/+)) mice, mice with the rds(-/-) (null) and rds(+/-) mutations, and mice with the P216L rds/peripherin mutation on the rds(+/ ) background were maintained in the vivarium under identical husbandry conditions, and tissues were removed from each group for analysis at approximately 2 months of age. Fatty acid compositions of total lipids from plasma, red blood cells, liver, and ROS were determined by gas-liquid chromatography. ROS purity from each group was determined by SDS-PAGE with silver staining. The morphologic status of retinas representing each genotype was analyzed by light and electron microscopy. RESULTS: There was no difference between rds(+/-), P216L on rds(+/-), and rds(+/+) (control) animals in the fatty acid composition of plasma, expressed as relative mole percent or as nanomoles fatty acid per milliliter of plasma. Small but statistically significant differences were found in 18:0 and C-22 polyunsaturated fatty acids of red blood cells. In the liver, the control animals had higher levels of 20:4n-6. In contrast, the ROS of control animals had levels of DHA that were 1.4 times that of ROS from either rds(+/-) or P216L on rds(+/-) mice of the same age. The reduction in DHA was not accompanied by an increase in 22:5n-6, which always occurs in neural tissues of animals deprived of n-3 fatty acids. SDS-PAGE of the three ROS membrane preparations showed that they were of identical purity. CONCLUSIONS: Mice heterozygous for the spontaneous rds/peripherin mutation or mice carrying the P216L mutation on this heterozygous background have a statistically significant reduction of DHA in their ROS membranes. The authors propose that reduction in DHA is an adaptive response to metabolic stress caused by the mutation. PMID- 11431434 TI - Overexpression of H- and L-ferritin subunits in lens epithelial cells: Fe metabolism and cellular response to UVB irradiation. AB - PURPOSE: To determine the effect of changes in ferritin subunit makeup on Fe metabolism and the resistance of lens epithelial cells (LEC) to photo-oxidative stress. METHODS: Cultured canine LEC were transiently transfected with pTargeT mammalian expression vector containing the whole coding sequence of H- or L-chain cDNA. The subunit composition of newly synthesized ferritin was analyzed by metabolic labeling and SDS-PAGE electrophoresis. Total ferritin concentration was measured by ELISA: Fe uptake and incorporation into ferritin was determined by incubating transfected cells with (59)Fe-labeled transferrin followed by native PAGE electrophoresis. The effect of UV irradiation was assessed by cell count after exposure of transfected cells to UVB radiation. RESULTS: Transfected cells differentially expressed H- and L-ferritin chains from cDNA under the control of CMV promoter; overexpression of L-chain was much greater than that of H-chain. The expressed chains assembled into ferritin molecules under in vitro and in vivo condition. The ferritin of H-transfectants incorporated significantly more Fe than those of L-transfectants. The UVB irradiation reduced cell number of L transfectants by half, whereas H-chain transfectants were protected. CONCLUSIONS: Post-transfectional expression of ferritin H- and L-chains in LEC appears to be regulated differentially. Overexpression of L-chain ferritin did not have a major effect on cellular Fe distribution and did not protect LEC against UV irradiation, whereas overexpression of H-chain resulted in increased storage of Fe in ferritin and protected cells from UV damage. PMID- 11431435 TI - Tropicamide (1%): an effective cycloplegic agent for myopic children. AB - PURPOSE: To evaluate the cycloplegic effect of 1% tropicamide in myopic children and to determine whether its efficacy is associated with age, gender, iris color, ethnicity, magnitude of the refractive error, or latent error. METHODS: Four hundred sixty-nine children enrolled in the Correction of Myopia Evaluation Trial (COMET; a multicenter, randomized, double-masked clinical trial evaluating the rate of progression of juvenile-onset myopia in children wearing progressive addition versus single-vision lenses) were given 1 drop of proparacaine in each eye followed 1 minute later by 1 drop of 1% tropicamide and then a second drop of 1% tropicamide 4 to 6 minutes later. Five accommodative responses to 20/100 letters located at 4 m and 33 cm were obtained in each eye with an autorefractor, 20 minutes after the second drop. Residual accommodation was calculated as the difference between the mean spherical equivalent responses obtained at the two distances. An examiner graded iris color, and ethnicity was reported by the children's parents or guardians. RESULTS: The mean residual accommodation was small: 0.38 +/- 0.41 diopters (D) in the right eye and 0.30 +/- 0.41 D in the left eye. Small but statistically significant differences in residual accommodation were associated with ethnicity, but not with any of the other factors. CONCLUSIONS: Tropicamide (1%) is an effective cycloplegic agent in myopic children. PMID- 11431436 TI - Determination of corneal asphericity after myopia surgery with the excimer laser: a mathematical model. AB - PURPOSE: To determine the theoretical change of corneal asphericity within the zone of laser ablation after a conventional myopia treatment, which conforms to Munnerlyn's paraxial formula and in which the initial corneal asphericity is not taken into consideration. METHODS: The preoperative corneal shape in cross section was modeled as a conic section of apical radius R(1) and shape factor p(1). A myopia treatment was simulated, and the equation of the postoperative corneal section within the optical zone was calculated by subtracting the ablation profile conforming to a general equation published by Munnerlyn et al. The apical radius of curvature r(2) of the postoperative profile was calculated analytically. The postoperative corneal shape was fitted by a conic section, with an apical radius equal to r(2) and a shape factor p(2) equal to the value that induced the lowest sum of horizontal residuals and the lowest sum of squared residuals. These calculations were repeated for a range of different dioptric treatments, initial shape factor values, and radii of curvature to determine the change of corneal asphericity within the optical zone of treatment. RESULTS: Analytical calculation of r(2) showed it to be independent of the initial preoperative shape factor p(1). The determination of p(2) was unambiguous, because the same value induced both the lowest sum of residuals and the lowest sum of the squared residuals. For corneas initially prolate (p(1) < 1), prolateness increased (p(2) < p(1) < 1), whereas for oblate corneas (p(1) > 1), oblateness increased (p(2) > p(1) > 1) within the treated zone after myopia treatment. This trend increased with the increasing magnitude of treatment and decreased with increasing initial apical radius of curvature R(1). CONCLUSIONS: After conventional myopic excimer laser treatment conforming to Munnerlyn's paraxial formula, the postoperative theoretical corneal asphericity can be accurately approximated by a best-fit conic section. For initially prolate corneas, there is a discrepancy between the clinically reported topographic trend to oblateness after excimer laser surgery for myopia and the results of these theoretical calculations. PMID- 11431437 TI - Role of tears in keratocyte loss after epithelial removal in mouse cornea. AB - PURPOSE: To study the role of tears in the death of keratocytes after epithelium removal in the mouse cornea. METHODS: In anesthetized mice, an approximately 1-mm circle of epithelium was removed from the center of the cornea, exposing the underlying stroma. In one group of animals, access of tears to the bare stroma was allowed-in vivo, by closing the eyelids, or ex vivo, by dropping tears from another animal onto the denuded stroma of an enucleated eyeball. In another group, tear access was denied-in vivo, by bathing the cornea continuously in saline or by keeping the lids open, or ex vivo, by rinsing the denuded cornea before incubating the enucleated eyeball. In a separate group, corneal epithelial debris from another mouse was placed on the bare stroma of an enucleated eyeball. The corneas were isolated, stained with a fluorescent nuclear dye, and observed en face in a wholemount preparation under a fluorescence microscope, to evaluate the distribution of intact nuclei across the entire depth of the stroma. RESULTS: Between 1.5 and 2 hours after exposure to tears, the nuclei of the anterior keratocytes under the area of epithelial debridement invariably degenerated. When they had been protected from the tears, however, no degeneration was observed. Epithelial debris applied on the bare stroma had no effect on the underlying keratocytes. CONCLUSIONS: Factors in tear fluid trigger keratocyte loss after removal of the epithelium in the mouse cornea. PMID- 11431438 TI - An x-ray diffraction investigation of corneal structure in lumican-deficient mice. AB - PURPOSE: The corneas of mice homozygous for a null mutation in lumican, a keratan sulfate-containing proteoglycan, are not as clear as normal. In the present study, mutant corneas were examined by synchrotron x-ray diffraction to see what structural changes might lie behind the loss of transparency. METHODS: X-ray diffraction patterns were obtained from the corneas of 6-month-old and 2-month old lumican-null and wild-type mice. Measured in each cornea were the average collagen fibril diameter, average collagen fibril spacing, and the level of order in the collagen array. RESULTS: The x-ray reflection arising from regularly packed collagen was well-defined on all x-ray patterns from 6-month-old wild-type corneas. Patterns from 6-month-old lumican-deficient corneas, however, contained interfibrillar reflections that were measurably more diffuse, a fact that points to a widespread alteration in the way the collagen fibrils are configured. The same distinction between mutant and wild-type corneas was also noted at 2-months of age. Average collagen fibril spacing was marginally higher in corneas of 6 month-old lumican-null mice than in corneas of normal animals. Unlike x-ray patterns from wild-type corneas, patterns from lumican-deficient corneas of both ages registered no measurable subsidiary x-ray reflection, evidence of a wider than normal range of fibril diameters. CONCLUSIONS: The spatial arrangement of stromal collagen in the corneas of lumican-deficient mice is in disarray. There is also a considerable variation in the diameter of the hydrated collagen fibrils. These abnormalities, seen at 2 months as well as 6 months of age, probably contribute to the reduced transparency. PMID- 11431439 TI - Integrity of epithelium and endothelium in organ-cultured human corneas. AB - PURPOSE: To examine components of the junctional complex and the actin cytoskeleton and the incidence of apoptosis in epithelium and endothelium of organ-cultured human corneas. METHODS: Human corneas, either organ-cultured for 1 to >28 days or excised directly from eyes stored in moist chambers, were stained with antibodies to ZO-1, vinculin, and caspase 3 coupled to FITC-conjugated secondary antibody. These markers were combined with rhodamine-phalloidin staining for F-actin and DAPI labeling for DNA. The corneas were examined by confocal microscopy. RESULTS: The depth of the epithelium was reduced during organ culture, but no changes were observed in the distribution of ZO-1 or vinculin, or in the F-actin cytoskeleton. The appearance of apoptotic epithelial cells positive for caspase 3 or with condensed DNA increased with time after 14 days in organ culture, but there was no correlation with donor age. ZO-1 and F actin staining patterns in endothelium were similarly undisturbed by organ culture, but apoptotic endothelial cells were only rarely seen and then only after >28 days in organ culture. CONCLUSIONS: Organ culture maintained the integrity of tight junctions and the actin cytoskeleton in epithelial and endothelial cell layers. Apoptosis was evident in epithelium but was observed rarely in the endothelium and then only after extended periods in organ culture. PMID- 11431440 TI - Glaucoma filtration surgery using amniotic membrane transplantation. AB - PURPOSE: To investigate the potential use of amniotic membrane transplantation (AMT) in the construction of glaucoma filtering blebs. METHODS: Twenty-four albino rabbits underwent glaucoma filtration surgery in one eye. In alternate cases, the conjunctival flap was replaced with AMT. Postoperative examination data were grouped into three time points. Six animals with AMT and six filtration surgery-controls were euthanatized at each of two postoperative time points, and tissue was obtained for histologic examination. Conjunctival biopsies were explanted for estimation of fibroblast outgrowth. RESULTS: Bleb formation was observed in all eyes, and amniotic membranes were epithelialized after 11.2 +/- 2.48 (mean +/- SD) days. Throughout the study IOPs were significantly lower in operated than unoperated fellow eyes. Between postoperative days 11 and 16 (the middle time point), the percentage IOP reduction in AMT eyes was significantly greater than in filtration surgery controls (P = 0.014), though not at other time points. Filtration surgery survival was significantly longer in the AMT group (22.3 +/- 3.8 days; mean +/- SE) than in "No AMT" controls (14.0 +/- 1.6 days; P = 0.035). In tissue culture, significantly less fibroblast outgrowth occurred from AMT explants when compared with unoperated conjunctiva (P = 0.01) between postoperative days 3 and 9 (the early time point). Amniotic membrane transplants were intact on histologic examination after 14 days but were associated with considerable granulomatous inflammation. After 36 days, the ocular surfaces remained clinically intact, but lysis of AMT was noted histologically. CONCLUSIONS: AMT exhibits potential as an alternative tissue to conjunctiva in the construction of glaucoma filtration blebs. The healing response as demonstrated by fibroblast outgrowth is retarded when compared with conventional conjunctival closure. The improvement in bleb survival must be weighed against the potential for complications related to delayed healing. In rabbits, human amniotic membrane elicited a late xenograft reaction, leading to granulomatous inflammation and dissolution of the membrane. PMID- 11431441 TI - Glucocorticoid induction of the glaucoma gene MYOC in human and monkey trabecular meshwork cells and tissues. AB - PURPOSE: To examine the intracellular and extracellular expression of myocilin in the human and primate trabecular meshwork (TM) in the presence and absence of glucocorticoids. METHODS: Myocilin expression was examined in cultured human TM cells by Northern blot analysis and myocilin antibody-mediated immunoprecipitation. Myocilin expression was quantified using high-resolution two dimensional polyacrylamide gel electrophoresis of radiolabeled proteins from human TM cells, TM tissue explants, and perfused human anterior segments cultured with and without dexamethasone (DEX) for 14 to 21 days, as well as TM tissue from pigtailed monkeys treated orally for 1 year with cortisone acetate. Immunofluorescence with anti-myocilin antibodies was used to localize cellular and extracellular expression of myocilin in cultured human TM cells. RESULTS: Glucocorticoid treatment caused a significant induction of myocilin mRNA, a tetrad of cell-associated proteins, and 8 to 20 secreted proteins (molecular mass [M(r)] 56 and 59 kDa and isoelectric point [pI] 5.2 and 5.3) in some, but not all the cultured human TM cells and explanted tissues. Western immunoblot analysis using anti-myocilin peptide antibodies identified these proteins as encoded by the MYOC gene. There was significant induction of the myocilin proteins in three perfusion-cultured human eyes, in which DEX-induced elevated intraocular pressure developed. Monkeys treated 1 year with cortisol acetate showed steroid glaucoma like morphologic changes in the TM that correlated with the induction of myocilin in the TM. Immunofluorescence analysis of cultured TM cells localized myocilin intracellularly in discrete perinuclear and cytoplasmic vesicular deposits as well as extracellularly on the cell surface associated with the extracellular matrix. In several DEX-treated TM cell lines, there were significant levels of myocilin secreted into the media. Enzymatic deglycosylation of proteins in the TM media converted the higher molecular weight isoforms of myocilin (approximately 57 kDa) to the lower molecular weight isoforms ( approximately 55 kDa). CONCLUSIONS: Although the function of myocilin is unknown, induction of these TM proteins was found in eyes in which glucocorticoid-induced ocular hypertension developed. Therefore, myocilin may play an important pathogenic role in ocular hypertension in addition to its role in certain forms of POAG. PMID- 11431442 TI - Reduced TIGR/myocilin protein in the monkey ciliary muscle after topical prostaglandin F(2alpha) treatment. AB - PURPOSE: Mutations in the trabecular meshwork inducible glucocorticoid response (TIGR) gene, also known as myocilin, have recently been linked to some forms of glaucoma. Recent studies have shown that TIGR protein also is expressed in the ciliary muscle. Because uveoscleral outflow, which traverses the ciliary muscle, is increased by prostaglandins (PGs), the present study assessed whether topical PGs alter the amount of TIGR protein within the ciliary muscle. METHODS: Vehicle was topically applied to one eye, and 2 microg PGF(2alpha)-isopropyl ester (PGF(2alpha)-IE) was applied to the other eye of cynomolgus monkeys twice daily for 5 days. Pressure reductions of 5 mm Hg in the PGF(2alpha)-IE-treated eyes were confirmed. The eyes were then fixed and paraffin sections were cut from each eye. The distribution of TIGR protein in the ciliary muscle was determined by confocal scanning laser microscopy. Additional sections were immunostained with a polyclonal antibody to recombinant TIGR protein or with a polyclonal antibody to a synthetic peptide corresponding to the leucine zipper region within the TIGR protein. Staining intensity in the ciliary muscle was assessed by measuring optical density (OD) along two line segments overlying the ciliary muscle, by using a high-resolution imaging densitometer. RESULTS: TIGR protein immunoreactivity was observed in ciliary muscle fibers throughout the ciliary muscle. Extracellular TIGR immunoreactivity colocalized with collagen type IV immunoreactivity. Intracellular staining also was present. Immunoreactivity was less intense in the sections from the PGF(2alpha)-IE-treated eyes compared with the vehicle-treated eyes. This was reflected in the reduction of mean OD scores in each monkey. Overall, the reduction of mean OD scores in the treated eyes was 42.1% +/- 9.9% (P < 0.005) with the anti-recombinant TIGR antibody and 27.3% +/- 10.4% with the anti-TIGR peptide antibody (P < 0.005). CONCLUSIONS: TIGR protein immunoreactivity was present both intracellularly and extracellularly in the ciliary muscle of the cynomolgus monkey. This suggests that extracellular TIGR protein is in contact with aqueous humor in the uveoscleral outflow pathway. Moreover, IOP-lowering topical PGF(2alpha)-IE treatment decreases the amount of TIGR protein in the ciliary muscle. PMID- 11431443 TI - TNF-alpha and TNF-alpha receptor-1 in the retina of normal and glaucomatous eyes. AB - PURPOSE: To determine the expression and localization of tumor necrosis factor (TNF)-alpha and TNF-alpha receptor-1 in the retina of normal and glaucomatous eyes. METHODS: Using immunohistochemistry and in situ hybridization, retinal expression and localization of TNF-alpha and TNF-alpha receptor-1 were studied in retina sections from 20 eyes of donors with glaucoma, and 20 eyes of age-matched normal donors. RESULTS: According to immunohistochemistry, the intensity of the immunostaining and the number of labeled cells for TNF-alpha or its receptor were greater in retina sections of glaucomatous eyes than in control eyes of age matched normal donors. In situ hybridization showed that mRNA signals for TNF alpha or TNF-alpha receptor-1 were similarly more intense in glaucomatous eyes than in age-matched control eyes. Both protein and mRNA of TNF-alpha or TNF-alpha receptor-1 were predominantly localized to the inner retinal layers. Double immunofluorescence labeling demonstrated that retinal immunostaining for TNF alpha was predominantly positive in the glial cells, whereas immunostaining for TNF-alpha receptor-1 was mainly positive in the retinal ganglion cells. CONCLUSIONS: Upregulation of TNF-alpha and its receptor-1 in glaucomatous retina suggest that TNF-alpha-mediated cell death is involved in the neurodegeneration process of glaucoma. PMID- 11431444 TI - Cationic ferritin changes outflow facility in human eyes whereas anionic ferritin does not. AB - PURPOSE: To determine the effect of charged moieties within the outflow pathway on aqueous outflow facility in human eyes. METHODS: After baseline facility measurement in human eye bank eyes (n = 10 pairs), one eye of each pair received anterior chamber exchange and continued perfusion with medium containing 10 mg/ml cationic ferritin. Contralateral eyes were treated in a similar manner with anionic ferritin (10.0 or 102 mg/ml). Eyes were fixed by anterior chamber exchange and perfusion with universal fixative at 8 mm Hg (corresponding to a physiologic pressure of 15 mm Hg in vivo) and examined by transmission electron microscopy. In a second series of human eyes (n = 8 pairs), facility was measured before and after anterior chamber exchange, with a solution containing 0.1 U/ml neuraminidase. RESULTS: Perfusion of eyes with anionic ferritin at either 10.0 or 102 mg/ml caused a negligible 2% increase in facility, whereas cationic ferritin perfusion reduced facility by 66% (P < 0.00001). Perfusion with fixative reduced facility by approximately 60% in both cationic and anionic ferritin-perfused eyes, relative to facilities after perfusion with ferritin. Transmission electron microscopy showed that the distribution of ferritin was segmentally variable. Cationic ferritin consistently labeled the luminal surface of the inner wall of Schlemm's canal, and variably labeled the juxtacanalicular connective tissue (JCT) and trabecular beam surfaces. Anionic ferritin was more prominent in the JCT and intertrabecular spaces and less so on the luminal surface of Schlemm's canal. By scanning electron microscopy, cationic ferritin was seen to accumulate at intercellular margins of the inner wall. Neuraminidase perfusion had no significant effect on outflow facility. CONCLUSIONS: Cationic ferritin reduces outflow facility, presumably by binding to negatively charged sites in the outflow pathway. A possible mechanism is partial or complete blockage of intercellular clefts in the inner wall of Schlemm's canal by the ferritin that accumulates on the luminal surface of the inner wall. Although they are possible targets for ferritin binding, sialyl residues themselves seem to have little direct effect on outflow facility. Our data indicate that positively charged molecules, especially if they can interact with inner wall pores, have the potential to markedly alter outflow facility. PMID- 11431445 TI - Expression of aquaporin-1 in human trabecular meshwork cells: role in resting cell volume. AB - PURPOSE: Drainage of aqueous humor from the human eye appears dependent on intracellular volume of trabecular meshwork (TM) cells, the predominant cell type of the human outflow pathway. Thus, the modulation of water and solute flux across the plasma membrane of TM cells is predicted to be an important factor in regulating outflow facility. Aquaporin (AQP)-1 is a hexahelical integral membrane protein that functions as a regulated channel for water and cations in fluid secreting and -absorbing tissues. AQP1 is present in many tissues of the human eye, including the TM; however, its role in outflow facility is unknown. The purpose of the present study was twofold: to evaluate the prospect of manipulating AQP1 protein levels in TM cells using sense and antisense mRNA and to investigate the functional role of AQP1 in TM cells. METHODS: An adenovirus (AV) expression system was used to alter AQP1 protein levels. AQP1 protein expression was monitored using immunoblot analysis, and resting cell volume was measured by forward light scatter, electronic cell sizing, and [(14)C] sucrose/urea equilibration. Permeability of TM monolayers to [(14)C]-sucrose was also assessed as an indirect evaluation of cell volume. RESULTS: AV-mediated gene transfer of AQP1 cDNA to TM cells resulted in a titer-dependent increase in recombinant AQP1, whereas transfer of antisense cDNA decreased native AQP1 protein by 71.7% +/- 5.5% (P < 0.01) after 5 days. A novel finding of this study is that mean resting volumes of AQP1(s) AV-infected TM cells in suspension were 8.7% +/- 3.0% greater (P < 0.05) than control cells. Conversely, AQP1 antisense (as) AV-infected cells had resting volumes 7.8% +/- 2.9% less than control cells (P < 0.05). Similar effects of AQP1 expression on resting cell volume were observed in TM monolayers. Consistent with this finding, paracellular permeability of AQP1(s) AV-infected TM monolayers to [(14)C]-sucrose decreased by 8.0% +/- 1.4% (P < 0.001). CONCLUSIONS: In addition to influencing the osmotic permeability of TM plasma membranes, the level of AQP1 protein expression influences resting intracellular volume and thus paracellular permeability of TM cell monolayers in vitro. These data suggest that AQP1 expression may affect outflow facility in vivo. PMID- 11431446 TI - The antiviral resistance and replication of cidofovir-resistant adenovirus variants in the New Zealand White rabbit ocular model. AB - PURPOSE: To determine the antiviral resistance of three cidofovir (CDV)-resistant variants of adenovirus type 5 (Ad5) and their ability to replicate in the New Zealand White rabbit ocular model. METHODS: Rabbits were inoculated topically in both eyes with the CDV-resistant variants R1, R2, and R3, and the Ad5 parental strain. On day 1, rabbits from each virus inoculation were divided into two topical treatment groups: 0.5% CDV and PBS control. Treatment was administered twice daily in both eyes for 7 days. All eyes were cultured for virus on days 0, 1, 3, 4, 5, 7, 9, 11, and 14. Using viral outcome parameters, CDV resistance was determined for each virus by comparing each CDV-treated virus group to its respective PBS control, and altered pathogenesis was assessed by comparing viral replication in the PBS control groups of the Ad5 parent and the three resistant variants. RESULTS: Topical 0.5% CDV treatment demonstrated significant antiviral inhibitory activity in the Ad5 parental group (e.g., reduced total Ad5-positive cultures, reduced daily Ad5-positive cultures on days 5, 9, 11, and 14, and duration of ocular shedding), but had no effect on the three CDV-resistant variants. There were no significant differences in pathogenicity between the Ad5 parent and the CDV-resistant variants. CONCLUSIONS: The Ad5 variants R1, R2, and R3 were resistant to topical treatment with 0.5% cidofovir in the rabbit ocular model. However, the acquisition of CDV resistance did not alter the replication of the three Ad5 CDV variants on the rabbit eye. PMID- 11431447 TI - Chlamydial antibodies in patients with previous acute anterior uveitis. AB - PURPOSE: To determine the prevalence of antibodies to Chlamydia pneumoniae, C. trachomatis, and C. pneumoniae heat shock protein (Cpn Hsp60) in patients with acute anterior uveitis (AAU) and in sex- and age-matched healthy control subjects. METHODS: Altogether 64 patients with previous AAU were examined at the Helsinki University Eye Hospital from September through December 1999. Serum specimens from the patients and sex- and age-matched healthy control subjects were tested for antibodies to C. pneumoniae and C. trachomatis by a specific microimmunofluorescence test and for antibodies to Cpn Hsp60 by enzyme immunoassay (EIA). RESULTS: The prevalence of antibodies to C. pneumoniae (69% vs. 72%) and C. trachomatis (11% vs. 6%) did not differ significantly between the patients and control subjects, nor did the level of IgG antibodies to Cpn Hsp60 (median EIA unit, 65 vs. 48). The levels of IgA antibodies to Cpn Hsp60 were significantly higher in the patients with AAU than in the control subjects (median EIA unit, 18 vs. 10; two-tailed Wilcoxon signed rank test, P = 0.0001). CONCLUSIONS: The high frequency of IgA antibodies to Cpn Hsp60 in patients with past AAU indicates that such patients may have persisting or recurrent infections due to C. pneumoniae. This finding suggests that C. pneumoniae may play a role in the pathogenesis of AAU. PMID- 11431448 TI - Dynamics of donor cell persistence and recipient cell replacement in orthotopic corneal allografts in mice. AB - PURPOSE: To determine the extent to which donor cells persist and recipient cells repopulate each of the three cell layers of orthotopic corneal grafts in mice. METHODS: BALB/c, C57BL/6, and enhanced green fluorescence protein (EGFP) transgenic mice (B6 background) were used as donors and recipients for orthotopic syngeneic and allogeneic corneal grafts. Graft-bearing eyes were harvested at 5, 10, 15, 28, and 56 days, stained with propidium iodide, and observed (layer by layer) by confocal microscopy. Bone marrow-derived cells in the grafts were assessed immunohistochemically. RESULTS: Donor epithelium was totally replaced by recipient epithelial cells within 15 days in both syngeneic and allogeneic grafts, whereas donor stromal keratocytes and endothelial cells were retained virtually intact in syngeneic grafts and in accepted allografts. In rejected allografts, neither donor-derived keratocytes nor endothelial cells were detected, and, instead, recipient-derived stromal fibroblasts, neovessels, and infiltrating leukocytes were heavily represented. The posterior surface of rejected grafts was devoid of corneal endothelium and was covered incompletely with bone marrow-derived cells of recipient origin. CONCLUSIONS: Whereas in mice graft-derived epithelium is largely irrelevant to corneal allograft outcome, persistence of donor-derived endothelium and keratocytes correlates perfectly with graft acceptance. Recipient endothelium is incapable of covering the posterior surface of accepted or rejected corneal grafts, whereas bone marrow derived cells of recipient origin come to occupy this site in rejected grafts. PMID- 11431449 TI - Binding of dexamethasone by alpha-crystallin. AB - PURPOSE: Long-term steroid therapy is a known risk factor for the development of posterior subcapsular cataract. Previous work in this laboratory has found soluble lens proteins to bind dexamethasone, but this binding is not due to a glucocorticoid receptor. This study was undertaken to identify the soluble protein or proteins involved in lens glucocorticoid binding. METHODS: Bovine lens extract was incubated with 5.2 x 10(-)(8) M [(3)H]-dexamethasone for 3 hours, and the distribution of label assessed in the soluble and insoluble fractions after centrifugation. Soluble lens extract was fractionated using gel permeation chromatography to isolate and identify proteins involved in the binding. Total lens proteins, high-molecular-weight proteins, or alpha-crystallin were exposed to dexamethasone and the protein bound steroid measured after separation of free and bound ligand on a gel chromatography column. Scatchard analysis was used to determine dexamethasone-binding parameters. Sequence comparisons between bovine alphaA- and alphaB-crystallins and glucocorticoid-binding proteins were performed using a sequence-alignment program. RESULTS: Of the total dexamethasone bound in lens extract, soluble proteins were found to account for 52%. The majority of the soluble protein-bound dexamethasone coeluted with the high-molecular-weight proteins that consisted mainly of alpha-crystallin. Binding studies with isolated proteins showed that alpha-crystallin accounted for more than 98% of total soluble dexamethasone binding in the lens. Scatchard analysis of steroid binding showed it to be a nonspecific partitioning event. Sequence comparisons between alphaA- and alphaB-crystallins and various glucocorticoid-binding proteins showed the lens proteins to have three regions of sequence homology with yeast corticosteroid-binding protein. CONCLUSIONS: alpha-Crystallin is the principal soluble glucocorticoid binding protein in the lens. The steroid association is described by nonspecific partitioning and may be related to the unique structural characteristics of the protein. The nonspecific association with alpha-crystallin is not thought to be functional; however, it may aid in the increased covalent steroid modification observed for this protein. PMID- 11431450 TI - Lens lactate dehydrogenase inactivation after UV-B irradiation: an in vivo measure of UVR-B penetration. AB - PURPOSE: To elucidate the spatial distribution of inactivation of lactate dehydrogenase (LDH) in ultraviolet-B radiation (UVR-B)-exposed eyes. To determine the in vivo penetration depth of UVR-B in the lens. METHODS: LDH activity in cornea and lens was investigated with an enzyme histochemical technique. Thirty rats were exposed in vivo to UVR-B of approximately 300 nm, and the eyes were enucleated and frozen at 0, 2, and 6 hours after exposure. LDH activity in frozen sections was determined quantitatively in the corneal epithelium and four different regions in the lens. UVR-B penetration depth was estimated by using a calculated Lambertian absorption coefficient. RESULTS: The LDH activity was decreased in the cornea and the outer anterior lens cortex at all three time points. The average decrease in enzyme activity in the time range was 35% in the cornea and 20% in the outer anterior lens cortex. UVR-B inhibition of LDH was immediate and not dependent on an inflammatory reaction within the eye. Penetration depth, corresponding to 1/e(2) ( approximately 14%) residual UVR-B intensity, was 0.45 mm. CONCLUSIONS: UVR-B does not exhibit any significant effect on LDH activity in the major part of the lens, and this is attributed to the shallow penetration (0.45 mm) of UVR-B into the anterior parts of the lens. PMID- 11431451 TI - Intraocular pressure responses to the adenosine agonist cyclohexyladenosine: evidence for a dual mechanism of action. AB - PURPOSE: Previous studies have shown that adenosine agonists are effective in reducing intraocular pressure (IOP). However, the mechanism(s) responsible for this ocular hypotensive effect has not been established. This study evaluates the relative contribution of changes in aqueous flow and outflow facility associated with the ocular hypotensive response to the adenosine agonist cyclohexyladenosine (CHA). METHODS: New Zealand White rabbits were treated topically in one eye with the adenosine A(1) agonist CHA. Changes in IOP, aqueous flow, and total outflow facility at various times after CHA administration were then determined. RESULTS: These studies demonstrated that CHA produces a dose-related reduction in IOP. Analysis of the dose-response curve revealed an ED(50) and a Hill coefficient of 87 microg and 1.9, respectively. Aqueous flow measurements demonstrated that 1.5 hours after CHA administration, aqueous flow was reduced by 35%. However, by 3.5 hours postdrug, no significant change in aqueous flow was observed. Measurement of the outflow facility found no significant change in facility 1.5 hours after CHA administration. However, by 3.5 hours after CHA administration, outflow facility was significantly increased by 85%. CONCLUSIONS: These data demonstrate that the adenosine agonist CHA lowers IOP in a dose-related fashion. This hypotensive action results from an early reduction in aqueous flow followed by a subsequent increase in outflow facility. This dual mechanism of action is consistent with analysis of CHA dose-response curve, which indicates that the reduction in IOP induced this agonist's results from multiple mechanisms of action. PMID- 11431452 TI - Reliable measurement of mouse intraocular pressure by a servo-null micropipette system. AB - PURPOSE: To develop a reliable technique for measuring intraocular pressure (IOP) in the mouse. METHODS: An electrophysiologic approach-the servo-null micropipette system (SNMS)-for measuring hydrostatic pressure was adapted for the mouse eye. Fine-tipped (5 microm in diameter) micropipettes were advanced across the cornea with a piezoelectric micromanipulator, and the IOP was continuously monitored for up to 46 minutes. RESULTS: The micropipette tip was visualized in the anterior chamber. With the SNMS, the IOP of black Swiss outbred mice under ketamine anesthesia was 17.8 +/- 0.4 mm Hg, higher than values previously estimated in inbred mouse strains by a larger bore microneedle manometric technique. After withdrawal of the micropipette, a second penetration led to a similar level of IOP. Hypotonic solutions increased and hypertonic solutions decreased IOP. Drugs that decrease inflow (acetazolamide, timolol) or increase outflow facility (pilocarpine, latanoprost) in primates and humans lowered steady state IOP in the mouse. The transient initial increase in IOP produced by pilocarpine reported in other animals was also observed in the mouse. Xylazine-ketamine anesthesia lowered IOP substantially in comparison with systemic anesthesia with either ketamine or tribromoethanol alone. CONCLUSIONS: The SNMS is the first reliable, reproducible method for measuring mouse IOP. The mouse IOP is sensitive not only to drugs known to reduce aqueous humor inflow but also to drugs that increase aqueous humor outflow facility in the eyes of primates and humans. The development of the SNMS is an enabling step in the use of the mouse for glaucoma research, including molecular genetics, molecular pharmacology, and the search for novel antiglaucoma drugs. PMID- 11431453 TI - Two types of K+ currents modulated by arachidonic acid in bovine corneal epithelial cells. AB - PURPOSE: Fenamate sensitivity of the large-conductance K+ current in the corneal epithelium suggests that K+ transport could be modulated by arachidonic acid (AA) and/or its metabolites, which also regulate corneal epithelial migration. The main purpose of this study was to investigate AA-induced modulation of K+ currents expressed in the bovine corneal epithelium. METHODS: Freshly isolated bovine corneal epithelial cells were perfused with Ringer solution. Whole-cell currents were recorded by using either the conventional whole-cell-patch or the perforated-patch configuration. RESULTS: Two distinct types of K+ currents dominated the whole-cell current. The first was a voltage-gated K+ current that was inactivated completely by membrane depolarization. The inactivating voltage gated K+ current was largest in presumptive basal cells. The second was a noisy, sustained K+ current that was never inactivated and seemed to be a counterpart of the large-conductance K+ current reported in the rabbit corneal epithelium. External application of AA (5-20 microm) inhibited the inactivating voltage-gated K+ current and augmented the noisy, sustained K+ current. Identical dual modulation was induced by other fatty acids (e.g., palmitoleic acid) that are not substrates for enzymes in the AA cascade. CONCLUSIONS: An inactivating voltage gated K+ channel was identified for the first time in the corneal epithelium. AA and some fatty acids may directly activate the large-conductance K+ channel to augment its housekeeping functions in corneal epithelial cells. PMID- 11431454 TI - Age-related accumulation and spatial distribution of lipofuscin in RPE of normal subjects. AB - PURPOSE: To characterize the age-related accumulation of lipofuscin in a population of normal subjects, resolve differences in estimated accumulation rates obtained in previous studies, and characterize the spatial distribution of lipofuscin in the normal fundus. METHODS: Spectrophotometric measurements were made at the fovea and 7 degrees temporal to the fovea in 145 normal subjects (age range, 15-80 years). Spatial distribution along the four cardinal meridians was measured in selected subjects by both spectrophotometry and autofluorescence imaging. To minimize contributions of extraneous fluorophores, macular pigment, and melanin, all measurements used excitation at 550 nm, integrating emission between 650 and 750 nm. RESULTS: Lipofuscin fluorescence increased linearly until age 70, then declined. The rate of accumulation was significantly slower in the fovea than at the temporal site; accumulation rates in vivo were greater than previously observed in microscopic studies. Fluorescence was approximately 40% lower in the fovea than at 7 degrees eccentricity and was asymmetrically distributed around the fovea. The fluorescence was maximal at approximately 11 degrees temporally, approximately 7 degrees nasally, approximately 13 degrees superiorly, and approximately 9 degrees inferiorly. At the same eccentricity, fluorescence was always less along the inferior meridian than along any other. CONCLUSIONS: Light absorption by RPE melanin can explain differences between the in vivo and ex vivo estimates of the rate of lipofuscin accumulation. Declining fluorescence at old age may represent removal of atrophic RPE cells. The spatial distribution of lipofuscin generally matches that of rods and reflects, rather than predicts, the pattern of age-related loss of rod photoreceptors. PMID- 11431455 TI - Captopril improves retinal neovascularization via endothelin-1. AB - PURPOSE: The purpose of this study was to determine the effect of an angiotensin converting enzyme inhibitor, captopril, on oxygen-induced retinopathy (OIR) in the mouse. Endothelin-1 (ET-1) expression is assessed in a mouse model of OIR. METHODS: OIR was produced in C57BL6 mice. Captopril (0.5mg/kg/d SC) was given from P7 (post natal day 7) for 5 days. Retinopathy was assessed by a retinal scoring system and by quantification of extra retinal neovascular nuclei on retinal sections at P17 to P20. The expression of ET-1 was determined using a reverse transcriptase polymerase chain reaction. RESULTS: Pups treated with captopril during hyperoxia had a lower median retinopathy score of 4.5 (25th, 75th quartile: 3, 6.4) compared with animals exposed to hyperoxia alone with median score 9.5 (25th, 75th quartile: 7.1, 10.4; P < 0.001). The pups treated with captopril during hyperoxia had significant reduction in number of nuclei extending beyond the inner limiting membrane (15.8 +/- 16.7, mean +/- SD) when compared with the animals exposed to hyperoxia only (50.4 +/- 28.0; P < 0.01). ET 1 expression in the retina increased 4.1-fold from P7 to P12 and a 1.9-fold increase from P12 to P17. Overall, there was an 8-fold increase in ET-1 expression from P7 to P17. Hyperoxia increased ET-1 expression by 2.1-fold at P12 over room air-reared animals. At P17, there was a 2.9-fold increase in retinal ET 1 expression when compared with room air. At P17, there was a 6.2-fold suppression in ET-1 expression in captopril-treated animals when compared with the oxygen only-treated animals. CONCLUSIONS: Captopril reduces retinal neovascularization in a mouse model of oxygen-induced retinopathy. ET-1 expression is increased from P7 to P17, altered by hyperoxic exposure and relative hypoxic recovery and modulated by captopril in a mouse model of OIR. PMID- 11431456 TI - Macular pigment and lutein supplementation in retinitis pigmentosa and Usher syndrome. AB - PURPOSE: To determine macular pigment (MP) in patients with inherited retinal degeneration and the response of MP and vision to supplementation of lutein. METHODS: Patients with retinitis pigmentosa (RP) or Usher syndrome and normal subjects had MP optical density profiles measured with heterochromatic flicker photometry. Serum carotenoids, visual acuity, foveal sensitivity, and retinal thickness (by optical coherence tomography [OCT]) were quantified. The effects on MP and central vision of 6 months of lutein supplementation at 20 mg/d were determined. RESULTS: MP density in the patients as a group did not differ from normal. Among patients with lower MP, there was a higher percentage of females, smokers, and light-colored irides. Disease expression tended to be more severe in patients with lower MP. Inner retinal thickness by OCT correlated positively with MP density in the patients. After supplementation, all participants showed an increase in serum lutein. Only approximately half the patients showed a statistically significant increase in MP. Retinal nonresponders had slightly greater disease severity but were otherwise not distinguishable from responders. Central vision was unchanged after supplementation. CONCLUSIONS: Factors previously associated with lower or higher MP density in normal subjects showed similar associations in RP and Usher syndrome. In addition, MP in patients may be affected by stage of retinal disease, especially that leading to abnormal foveal architecture. MP could be augmented by supplemental lutein in many but not all patients. There was no change in central vision after 6 months of lutein supplementation, but long-term influences on the natural history of these retinal degenerations require further study. PMID- 11431457 TI - Phenotypic marker for early disease detection in dominant late-onset retinal degeneration. AB - PURPOSE: To define early disease expression in autosomal dominant late-onset retinal degeneration (L-ORD), a retinopathy that becomes symptomatic after age 50 and is characterized histopathologically by sub-RPE deposits. METHODS: Three families with L-ORD were included; two families had postmortem eye donors with retina-wide sub-RPE deposits. Six patients with severe visual loss (ages 62-93) were examined clinically, and 17 available individuals (ages 35-60) at a 50:50 risk to inherit L-ORD were also studied with dark adaptometry. A short-term trial of vitamin A at 50,000 IU/day was conducted in three members. Three-year follow up examinations were performed in a subset of members. RESULTS: Family 1 had 12 available members at risk. On initial examination, only one member had fundus abnormalities: yellow-white punctate lesions in the midperipheral fundus. Dark adaptation kinetics were abnormal in 6 of 12. The youngest age with an abnormality was 35. Family 2 had two available members at risk, both of whom had punctate fundus lesions and abnormal dark adaptation. Family 3 had three available members at risk. One had fundus lesions and abnormal dark adaptation, whereas the others had normal fundi and normal adaptometry. Vitamin A accelerated adaptation kinetics but not to normal rates. Three-year follow-up examinations demonstrated further slowing of adaptation kinetics, whereas rod and cone thresholds remained unchanged. CONCLUSIONS: Dark-adaptation abnormalities can precede symptoms and funduscopic signs of L-ORD by at least a decade. Short-term, high-dose vitamin A accelerates the kinetics of dark adaptation to a limited degree. The results contribute clues about early pathophysiology of this retinal degeneration and provide additional power for genetic mapping of the L-ORD locus. PMID- 11431458 TI - Effects of cholesterol and apolipoprotein E on retinal abnormalities in ApoE deficient mice. AB - PURPOSE: To examine the pathologic changes in the retina of apolipoprotein E (apoE)-deficient mice fed a high-cholesterol diet. METHODS: ApoE-deficient mice (ApoE) were maintained on either regular mouse chow (ApoE-R) or a high cholesterol diet (ApoE-C) for 25 weeks. Age-matched control C57BL/6J mice (C57) were also maintained on either regular mouse chow (C57-R) or a cholesterol containing diet (C57-C). Retinal function was assessed by dark-adapted electroretinography (ERG). The eyes were embedded, sectioned, and analyzed by histologic and immunohistochemical methods, as well as by light and transmission electron microscopy. RESULTS: After the 25-week feeding period, ERG tracings of ApoE-C mice revealed significant increases of a- and b-wave implicit times when compared with the C57-R group of mice. In addition, there were reductions in oscillatory potential (OP) amplitudes in the ApoE-C group. However, a- and b-wave amplitudes appeared to be unchanged among the four groups of mice. Light microscopic examination of the retinas showed that compared with control C57-R mice, ApoE-C mice had significantly lower cell numbers in the inner and outer nuclear layers (85.1% +/- 4.6%, P < 0.05 and 81.4% +/- 3.7%, P < 0.01 of C57-R controls, respectively). Transmission electron microscopy of apoE-deficient mice revealed cells of the inner nuclear layer with condensation of nuclear chromatin and perinuclear vacuolization in focal areas. Bruch's membrane was also found to be thicker, and its elastic lamina appeared disorganized and discontinuous. Immunohistochemistry demonstrated diminished or no immunoreactivity for carbonic anhydrase II and calretinin in the retinal layers of apoE-deficient mice. CONCLUSIONS: Overall, there were increasing abnormalities of retinal function and cellular morphology among the four groups of mice in the order of C57-R < C57-C < ApoE-R < ApoE-C. These findings suggest that apoE and/or cholesterol play an important role in retinal function. PMID- 11431459 TI - Identification and localization of retinal cystatin C. AB - PURPOSE: Cystatin C is a mammalian cysteine protease inhibitor, synthesized in various amounts by many kinds of cells and appearing in most body fluids. There are reports that it may be synthesized in the mammalian retina and that a cysteine protease inhibitor may influence the degradation of photoreceptor outer segment proteins. In the current study cystatin C was identified, quantitated, and localized in mouse, rat, and human retinas. METHODS: Enzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT PCR), DNA sequencing, Western blot analysis, and immunohistochemistry have been used on mouse, rat, and human retinas (pigment epithelium included). RESULTS: Cystatin C is present in high concentrations in the normal adult rat retina, as it is throughout its postnatal development. Its concentration increases to a peak at the time when rat pups open their eyes and then remains at a high level. It is mainly localized to the pigment epithelium, but also to some few neurons of varying types in the inner retina. Cystatin C is similarly expressed in normal mouse and human retinas. CONCLUSIONS: Cystatin C was identified and the localization described in the retinas of rat, mouse, and human using several techniques. Cystatin C is known to efficiently inactivate certain cysteine proteases. One of them, cathepsin S, is present in the retinal pigment epithelium and affects the proteolytic processing by cathepsin D of diurnally shed photoreceptor outer segments. Hypothetically, it appears possible that retinal cystatin C, given its localization to the pigment epithelium and its ability to inhibit cathepsin S, could be involved in the regulation of photoreceptor degradation. PMID- 11431460 TI - Distribution of transcript and protein isoforms of the synaptic glycoprotein neuroplastin in rat retina. AB - PURPOSE: To examine the expression and localization of the neuroplastins (np), two synapse-enriched members of the immunoglobulin (Ig) superfamily of cell adhesion molecules, in the developing and adult retina and optic nerve. METHODS: Expressions of the two isoforms np55 and np65 and carboxyl-terminal splice variants were investigated by immunocytochemistry, Western blot analysis, RT-PCR, and in situ hybridization. RESULTS: Immunoreactivity for both neuroplastins was confined to the two synaptic layers of the retina: the inner (IPL) and outer plexiform layer (OPL). Significant overlap was found in staining at synaptic structures with synaptophysin. A large proportion of immunoreactivity for both isoforms, however, was of perisynaptic origin. In situ hybridization studies were suggestive of a pre- and postsynaptic localization of np65 in the OPL. Transcripts for np55 were already present at birth in the inner retina, but the hybridization signals increased during postnatal development. Np65 transcripts and immunosignals appeared at later developmental ages, concomitant with synapse formation in the OPL. Several C-terminal neuroplastin cDNA clones harbor an insert of 12 bp, coding for four amino acids (DDEP) in the intracellular domain of neuroplastins. Splice isoforms containing the insert exhibited a developmental expression pattern similar to that of np55; however, both neuroplastins could harbor the C-terminal insert. Neuroplastins were also detected in optic nerve homogenates. RT-PCR and blockade of axonal transport by nerve crush confirmed transcript and protein expression in optic nerve tissue. CONCLUSIONS: The findings suggest a role for neuroplastins in cell adhesion in the plexiform layers during histogenesis, as well as in maintenance of connections between specific cellular structures. PMID- 11431461 TI - Diabetes-induced disruption of gap junction pathways within the retinal microvasculature. AB - PURPOSE: Microvascular damage caused by diabetes is a leading cause of visual loss. Identifying events early in the course of diabetic retinopathy may help in understanding and, perhaps, preventing this disorder. The hypothesis that cell-to cell communication within the retinal microvasculature may be affected soon after the onset of diabetes was tested. METHODS: Streptozotocin was used to induce diabetes in rats. To assess cell-to-cell coupling the gap junction-permeant tracer, Neurobiotin, was delivered via patch pipettes into pericytes located on microvessels freshly isolated from the retinas of diabetic and control animals. Subsequently, immunohistochemical methods revealed the extent of the intercellular spread of the tracer. Electrophysiological methods were also used to detect intercellular communication. RESULTS: In retinal microvessels of control rats, Neurobiotin spread hundreds of micrometers from the tracer-loaded pericytes. However, within days after the onset of diabetes, this cell-to-cell coupling was dramatically reduced. In contrast, microvessels of insulin-treated diabetic rats showed no significant loss of intercellular communication. Consistent with protein kinase C (PKC) playing a role in the diabetes-induced inhibition of gap junction pathways, exposure of microvessels to a PKC activator (phorbol myristate acetate) markedly reduced tracer coupling. CONCLUSIONS: Within retinal microvessels there is extensive cell-to-cell coupling, which is markedly reduced soon after the onset of streptozotocin-induced diabetes. The closure of gap junction pathways disrupts the multicellular organization of retinal microvessels and may contribute to vascular dysfunction. PMID- 11431462 TI - Epinephrine-induced increases in [Ca2+](in) and KCl-coupled fluid absorption in bovine RPE. AB - PURPOSE: To define the ionic basis for the apical epinephrine-induced increase of fluid absorption (J(V)) across isolated bovine RPE-choroid. METHODS: Epinephrine induced changes in RPE [Ca2+](in) levels were monitored with the ratioing dye fura-2. Transepithelial potential, resistance, and unidirectional fluxes of (36)Cl, (86)Rb (K substitute), and (22)Na were simultaneously determined in paired tissues from the same eye mounted in modified Ussing flux chambers. Radioisotopes (5-7 microCi) were added to the apical bath of one tissue and the basal bath of the other, and the appearance of label in the opposite bath was measured. RESULTS: Apical epinephrine (100 nM) transiently increased [Ca2+](in) by 153 +/- 78 nM. This increase was inhibited by the alpha(1)-adrenoreceptor antagonist prazosin (1 microM) and blocked by CPA(5 microM), an inhibitor of endoplasmic reticulum Ca2+-adenosine triphosphatases (ATPases). Apical epinephrine (100 nM) more than doubled the net Cl absorption rate, increased net K ((86)Rb) absorption by fivefold, and tripled net fluid absorption (J(V)), as predicted by isotonic coupling between ion and fluid transport. The epinephrine induced increases in ion and fluid transport were completely inhibited by apical bumetanide (100 microM). CONCLUSIONS: Epinephrine increased fluid absorption across bovine RPE by activating apical membrane alpha(1)-adrenergic receptors, increasing [Ca2+](in), and stimulating bumetanide-sensitive Na,K,2Cl uptake at the apical membrane and KCl efflux at the basolateral membrane. PMID- 11431463 TI - Inhibitory effects of neurocan and phosphacan on neurite outgrowth from retinal ganglion cells in culture. AB - PURPOSE: Neurocan and phosphacan are nervous tissue-specific chondroitin sulfate proteoglycans (CSPGs) that are highly expressed in postnatal rat retina. To elucidate potential roles of neurocan and phosphacan on neurite outgrowth from retinal ganglion cells (RGCs), in vitro experiments were conducted with purified RGCs. METHODS: Neurocan and phosphacan were purified from postnatal rat brain by DEAE-column chromatography and subsequent gel chromatography. RGCs were obtained from postnatal rat retinas by a two-step immunopanning procedure using an anti Thy 1,1 antibody and an anti-macrophage antibody. Neurite outgrowth from RGCs was examined on poly-L-lysine (PLL)-conditioned plates, and PLL-conditioned plates treated with neurocan or phosphacan. RESULTS: Compared with PLL-conditioned plates, neurocan and phosphacan inhibited neurite outgrowth from RGCs at 48 and 72 hours after seeding. When chondroitin sulfate side chains linked to the core proteins were digested by chondroitinase ABC, the inhibitory effect remained, indicating that the core proteins are related to the effect. Furthermore, the digestion of chondroitin sulfate side chains linked to phosphacan core protein significantly promoted the inhibitory effect of phosphacan on neurite outgrowth from RGCs. CONCLUSIONS: Neurocan and phosphacan, which are highly expressed in postnatal rat retina, inhibit neurite outgrowth from postnatal rat RGCs, indicating that these proteoglycans may be inhibitory factors against neurite outgrowth from RGCs during retinal development. PMID- 11431464 TI - PDGF-induced coupling of function with metabolism in microvascular pericytes of the retina. AB - PURPOSE: The aim of this study was to test the hypothesis that platelet-derived growth factor (PDGF)-BB regulates the physiology of retinal pericytes, which are contractile cells located on the abluminal surface of capillaries. The expression of PDGF-BB and its cognate receptor in retinal vessels suggests a vasoactive function. However, although endothelium-derived PDGF-BB appears vital for the development of pericyte-containing microvessels, its role in the mature vasculature remains uncertain. METHODS: Based on the premise that ion channels mediate the responses of pericytes to vasoactive signals, the perforated-patch configuration of the patch-clamp technique was used to determine the effect of PDGF-BB on the ionic currents and membrane potential of pericytes located on microvessels freshly isolated from the adult rat retina. Changes in pericyte calcium levels were monitored with the calcium indicator fluo-4. Differential interference contrast optics and image analysis software aided in assessing the effects of PDGF-BB on the lumens of isolated pericyte-containing microvessels. In some experiments, blockers of adenosine triphosphate (ATP) synthesis created chemical ischemia. RESULTS: Electrophysiological recordings from pericytes showed that PDGF-BB can activate nonspecific cation channels, chloride channels, and ATP sensitive potassium channels. The metabolic status of an isolated capillary determined which of these ion channels were activated by PDGF-BB and thereby whether the membrane potential decreased or increased, the cell calcium rose or fell, and the vessel lumen constricted or dilated. CONCLUSIONS: The ability of PDGF-BB to be a vasoconstrictor when energy supplies are ample and to be a vasodilator under ischemic conditions may provide an efficient mechanism to link capillary function to local metabolic needs. PMID- 11431465 TI - Development of a reading speed test for potential-vision measurements. AB - PURPOSE: Previous studies suggest that optimal reading speed is unaffected by cataract, yet is significantly reduced in age-related macular degeneration (ARMD ). This raises the question of whether a reading speed test could be developed to assess potential vision after cataract surgery. METHODS: Nineteen subjects with cataract, 15 with ARMD, and 13 control subjects with normal, healthy eyes read Bailey-Lovie word charts aloud, and subsequently, critical print size and optimal reading speed were calculated. Measurements were also taken with the charts in reversed-contrast polarity and after pupillary dilation. RESULTS: Although the subjects with cataract had reduced word acuity and increased critical print size, optimal reading speed was similar to that of the control group at a mean of approximately 100 wpm. Optimal reading speed in the subjects with ARMD was substantially worse (mean of 39 wpm). Reversing the contrast polarity of the charts slightly increased the word acuity and optimal reading speed of the subjects with cataract. CONCLUSIONS: The results suggest that optimal reading speed would be useful as a potential-vision test. The proposed test would use text size of at least 1.32 degrees (1.2 log minimum angle of resolution [logMAR]), and pupil dilation would be unnecessary. A reading test with black letters on a white background would be adequate, because charts with reversed contrast polarity made minimal difference in reading speed. PMID- 11431466 TI - The Drosophila U1 and U6 gene proximal sequence elements act as important determinants of the RNA polymerase specificity of small nuclear RNA gene promoters in vitro and in vivo. AB - Transcription of genes coding for metazoan spliceosomal snRNAs by RNA polymerase II (U1, U2, U4, U5) or RNA polymerase III (U6) is dependent upon a unique, positionally conserved regulatory element referred to as the proximal sequence element (PSE). Previous studies in the organism Drosophila melanogaster indicated that as few as three nucleotide differences in the sequences of the U1 and U6 PSEs can play a decisive role in recruiting the different RNA polymerases to transcribe the U1 and U6 snRNA genes in vitro. Those studies utilized constructs that contained only the minimal promoter elements of the U1 and U6 genes in an artificial context. To overcome the limitations of those earlier studies, we have now performed experiments that demonstrate that the Drosophila U1 and U6 PSEs have functionally distinct properties even in the environment of the natural U1 and U6 gene 5'-flanking DNAs. Moreover, assays in cells and in transgenic flies indicate that expression of genes from promoters that contain the "incorrect" PSE is suppressed in vivo. The Drosophila U6 PSE is incapable of recruiting RNA polymerase II to initiate transcription from the U1 promoter region, and the U1 PSE is unable to recruit RNA polymerase III to transcribe the U6 gene. PMID- 11431467 TI - Protein oxidation in G0 cells of Saccharomyces cerevisiae depends on the state rather than rate of respiration and is enhanced in pos9 but not yap1 mutants. AB - Immunodetection of protein carbonyl groups demonstrates that growth arrest elicited by carbon or nitrogen starvation causes an increased oxidation of proteins in Saccharomyces cerevisiae. Mutant analysis suggests that the response regulator Pos9p is involved in mitigating self-inflicted oxidative damages in G(0) cells, whereas Yap1p is primarily required in growing cells. The data also suggest that oxidation of target proteins is not a priori an effect of arrest of cell division or nutrient depletion and cannot be explained by the respiratory activity alone nor a high ratio of catabolic/anabolic activity in G(0) cells. Instead, we observed that starvation elicits a transition in the respiratory state (from phosphorylating to nonphosphorylating respiration) and that this transition is associated with a stepwise increase in protein oxidation. During carbon starvation, this transition and increase in oxidation occurs immediately as the carbon source is depleted, growth is arrested, and the respiratory rate falls drastically. In contrast, during nitrogen starvation and excess carbon the respiratory state transition and stepwise increase in protein oxidation are markedly delayed and occur long after the nitrogen source has been depleted and division and growth-arrested. Oxidation in G(0) cells could be enhanced by treating cells with low concentrations of antimycin A and attenuated with myxothiazol, indicating that protein oxidation is intimately linked to reactive oxygen species generated by semiquinones of the Q-cycle. Thus, the work presented suggests that the degree of coupling in the mitochondrial respiratory apparatus rather then the overall rate of respiration affects the degree of protein oxidation in nondividing yeast cells. PMID- 11431468 TI - Flavopiridol inactivates P-TEFb and blocks most RNA polymerase II transcription in vivo. AB - Flavopiridol (L86-8275, HMR1275) is a cyclin-dependent kinase (Cdk) inhibitor in clinical trials as a cancer therapy that has been recently shown to block human immunodeficiency virus Tat transactivation and viral replication through inhibition of positive transcription elongation factor b (P-TEFb). Flavopiridol is the most potent P-TEFb inhibitor reported and the first Cdk inhibitor that is not competitive with ATP. We examined the ability of flavopiridol to inhibit P TEFb (Cdk9/cyclin T1) phosphorylation of both RNA polymerase II and the large subunit of the 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) sensitivity-inducing factor and found that the IC(50) determined was directly related to the concentration of the enzyme. We concluded that the flavonoid associates with P-TEFb with 1:1 stoichiometry even at concentrations of enzyme in the low nanomolar range. These results indicate that the apparent lack of competition with ATP could be caused by a very tight binding of the drug. We developed a novel immobilized P-TEFb assay and demonstrated that the drug remains bound for minutes even in the presence of high salt. Flavopiridol remained bound in the presence of a 1000-fold excess of the commonly used inhibitor DRB, suggesting that the immobilized P-TEFb could be used in a simple screening assay that would allow the discovery or characterization of compounds with binding properties similar to flavopiridol. Finally, we compared the ability of flavopiridol and DRB to inhibit transcription in vivo using nuclear run-on assays and concluded that P-TEFb is required for transcription of most RNA polymerase II molecules in vivo. PMID- 11431469 TI - Cross-talk between the ERK and p70 S6 kinase (S6K) signaling pathways. MEK dependent activation of S6K2 in cardiomyocytes. AB - The alpha(1)-adrenergic agonist phenylephrine (PE) and insulin each stimulate protein synthesis in cardiomyocytes. Activation of protein synthesis by PE is involved in the development of cardiac hypertrophy. One component involved here is p70 S6 kinase 1 (S6K1), which lies downstream of mammalian target of rapamycin, whose regulation is thought to involve phosphatidylinositol 3-kinase and protein kinase B (PKB). S6K2 is a recently identified homolog of S6K1 whose regulation is poorly understood. Here we demonstrate that in adult rat ventricular cardiomyocytes, PE and insulin each activate S6K2, activation being 3.5- and 5-fold above basal, respectively. Rapamycin completely blocked S6K2 activation by either PE or insulin. Three different inhibitors of MEK1/2 abolished PE-induced activation of S6K2 whereas expression of constitutively active MEK1 activated S6K2, without affecting the p38 mitogen-activated protein kinase and JNK pathways, indicating that MEK/ERK signaling plays a key role in regulation of S6K2 by PE. PE did not activate PKB, and expression of dominant negative PKB failed to block activation of S6K2 by PE, indicating PE-induced S6K2 activation is independent of PKB. However, this PKB mutant did partially block S6K2 activation by insulin, indicating PKB is required here. Another hypertrophic agent, endothelin 1, also activated S6K2 in a MEK-dependent manner. Our findings provide strong evidence for novel signaling connections between MEK/ERK and S6K2. PMID- 11431470 TI - Regulation of ubiquitination and degradation of p53 in unstressed cells through C terminal phosphorylation. AB - Previously, we found that the protein kinase C (PKC) inhibitor H7 stimulates p53 to accumulate in a form incapable of inducing transcription from p53-dependent promoters. We concluded that H7 inhibits constitutive C-terminal phosphorylation of p53, which regulates its turnover in unstressed cells. We now show that p53 and its inhibitor MDM2 (HDM2 in human cells) are together in the nuclei of H7 treated cells and can be co-immunoprecipitated. Despite this association of p53 with the ubiquitin ligase MDM2, ubiquitinated p53 was not detected in H7-treated cells. Furthermore, co-treatment with H7 and the proteosome inhibitor LLnL prevented the accumulation of ubiquitinated p53 that was observed in cells treated solely with LLnL. In addition, treatment of cells with the PKC activator phorbol ester stimulated the ubiquitination of p53 and reduced its ability to accumulate after stress. H7 did not induce the phosphorylation of human p53 on Ser-15 (Ser-18 in mouse protein), a modification that occurs in response to DNA damage and leads to the release of MDM2 and to transactivation by p53. We conclude that phosphorylation of the C-terminal domain of p53 by PKC increases its ubiquitination and degradation in unstressed cells. PMID- 11431471 TI - Asymmetric interactions between the acidic P1 and P2 proteins in the Saccharomyces cerevisiae ribosomal stalk. AB - The Saccharomyces cerevisiae ribosomal stalk is made of five components, the 32 kDa P0 and four 12-kDa acidic proteins, P1alpha, P1beta, P2alpha, and P2beta. The P0 carboxyl-terminal domain is involved in the interaction with the acidic proteins and resembles their structure. Protein chimeras were constructed in which the last 112 amino acids of P0 were replaced by the sequence of each acidic protein, yielding four fusion proteins, P0-1alpha, P0-1beta, P0-2alpha, and P0 2beta. The chimeras were expressed in P0 conditional null mutant strains in which wild-type P0 is not present. In S. cerevisiae D4567, which is totally deprived of acidic proteins, the four fusion proteins can replace the wild-type P0 with little effect on cell growth. In other genetic backgrounds, the chimeras either reduce or increase cell growth because of their effect on the ribosomal stalk composition. An analysis of the stalk proteins showed that each P0 chimera is able to strongly interact with only one acidic protein. The following associations were found: P0-1alpha.P2beta, P0-1beta.P2alpha, P0-2alpha.P1beta, and P0-2beta.P1alpha. These results indicate that the four acidic proteins do not form dimers in the yeast ribosomal stalk but interact with each other forming two specific associations, P1alpha.P2beta and P1beta.P2alpha, which have different structural and functional roles. PMID- 11431472 TI - Rim1 and rabphilin-3 bind Rab3-GTP by composite determinants partially related through N-terminal alpha -helix motifs. AB - Rim1 is a protein of the presynaptic active zone, the area of the plasma membrane specialized for neurotransmitter exocytosis, and interacts with Rab3, a small GTPase implicated in neurotransmitter vesicle dynamics. Here, we have studied the molecular determinants of Rim1 that are responsible for Rab3 binding, employing surface plasmon resonance and recombinant, bacterially expressed Rab3 and Rim1 proteins. A site that binds GTP- but not GDP-saturated Rab3 was localized to a short alpha-helical sequence near the Rim1 N terminus (amino acids 19-55). Rab3 isoforms A, C, and D were bound with similar affinities (K(d) = 1-2 microm). Low affinity binding of Rab6A-GTP was also observed (K(d) = 16 microm), whereas Rab1B, -5, -7, -8, or -11A did not bind. Adjacent sequences up to amino acid 387, encompassing differentially spliced sequences, the zinc finger module, and the SGAWFF motif of Rim1, did not significantly contribute to the strength or the specificity of Rab3 binding, whereas a point mutation within the helix (R33G) abolished binding. This Rab3 binding site of Rim1 is reminiscent of the N terminal alpha-helix that is part of the Rab3-binding region of rabphilin-3, and indeed we observed low affinity, specific binding of Rab3A (K(d) on the order of magnitude of 10-100 microm) to this region of rabphilin-3 alone (amino acids 40 88), whereas additional sequences up to amino acid 178 are needed for high affinity Rab3A binding to rabphilin-3 (K(d) = 10-20 nm). In contrast, an N terminal alpha-helix motif in aczonin, with sequence similarity to the Rab3 binding site of Rim1, did not bind Rab3A, -C, or -D or several other Rab proteins. These results were qualitatively confirmed in pull-down experiments with native, prenylated Rab3 from brain lysate in Triton X-100. Munc13 bound to the zinc finger domain of Rim1 but not to the rabphilin-3 or aczonin zinc fingers. Pull-down experiments from brain lysate in the presence of cholate as detergent detected binding to downstream Rim1 sequences, between amino acids 56 and 387, of syntaxin and of Rab3. The latter, however, was inhibited rather than stimulated by GTP. PMID- 11431473 TI - Rich, a rho GTPase-activating protein domain-containing protein involved in signaling by Cdc42 and Rac1. AB - A previously unidentified Rho GTPase-activating protein (GAP) domain-containing protein was found in a yeast two-hybrid screen for cDNAs encoding proteins binding to the Src homology 3 domain of Cdc42-interacting protein 4 (CIP4). The protein was named RICH-1 (RhoGAP interacting with CIP4 homologues), and, in addition to the RhoGAP domain, it contained an N-terminal domain with endophilin homology and a C-terminal proline-rich domain. Transient transfections of RICH-1 indicated that it bound to CIP4 in vivo, as shown by co-immunoprecipitation experiments, as well as co-localization assays. In vitro assays demonstrated that the RhoGAP domain of RICH-1 catalyzed GTP hydrolysis on Cdc42 and Rac1, but not on RhoA. Ectopic expression of the RhoGAP domain as well as the full-length protein interfered with platelet-derived growth factor BB-induced membrane ruffling, but not with serum-induced stress fiber formation, further emphasizing the notion that, in vivo, RICH-1 is a GAP for Cdc42 and Rac1. PMID- 11431474 TI - Ten ERK-related proteins in three distinct classes associate with AP-1 proteins and/or AP-1 DNA. AB - We have identified seven ERK-related proteins ("ERPs"), including ERK2, that are stably associated in vivo with AP-1 dimers composed of diverse Jun and Fos family proteins. These complexes have kinase activity. We designate them as "class I ERPs." We originally hypothesized that these ERPs associate with DNA along with AP-1 proteins. We devised a DNA affinity chromatography-based analytical assay for DNA binding, the "nucleotide affinity preincubation specificity test recognition" (NAPSTER) assay. In this assay, class I ERPs do not associate with AP-1 DNA. However, several new "class II" ERPs do associate with DNA. p41 and p44 are ERK1/2-related ERPs that lack kinase activity and associate along with AP-1 proteins with AP-1 DNA. Class I ERPs and their associated kinase activity thus appear to bind AP-1 dimers when they are not bound to DNA and then disengage and are replaced by class II ERPs to form higher order complexes when AP-1 dimers bind DNA. p97 is a class III ERP, related to ERK3, that associates with AP-1 DNA without AP-1 proteins. With the exception of ERK2, none of the 10 ERPs appear to be known mitogen-activated protein kinase superfamily members. PMID- 11431475 TI - Crystal structures of 1-aminocyclopropane-1-carboxylate (ACC) synthase in complex with aminoethoxyvinylglycine and pyridoxal-5'-phosphate provide new insight into catalytic mechanisms. AB - The structures of tomato 1-aminocyclopropane-1-carboxylate synthase (ACS) in complex with either cofactor pyridoxal-5'-phosphate (PLP) or both PLP and inhibitor aminoethoxyvinylglycine have been determined by x-ray crystallography. The structures showed good conservation of the catalytic residues, suggesting a similar catalytic mechanism for ACS and other PLP-dependent enzymes. However, the proximity of Tyr152 to the C-gamma-S bond of model substrate S-adenosylmethionine implies its critical role in the catalysis. The concerted accomplishment of catalysis by cofactor PLP and a protein residue, as proposed on the basis of the ACS structures in this paper, may represent a general scheme for the diversity of PLP-dependent catalyses. PLP-dependent enzymes have been categorized into four types of folds. A structural comparison revealed that a core fragment of ACS in fold type I is superimposable over tryptophan synthase beta subunit in fold type II and mouse ornithine decarboxylase in fold type III, thus suggesting a divergent evolution of PLP-dependent enzymes. PMID- 11431476 TI - The autocatalytic release of a putative RNA virus transcription factor from its polyprotein precursor involves two paralogous papain-like proteases that cleave the same peptide bond. AB - The largest replicative protein of coronaviruses is known as p195 in the avian infectious bronchitis virus (IBV) and p210 (p240) in the mouse hepatitis virus. It is autocatalytically released from the precursors pp1a and pp1ab by one zinc finger-containing papain-like protease (PLpro) in IBV and by two paralogous PLpros, PL1pro and PL2pro, in mouse hepatitis virus. The PLpro-containing proteins have been recently implicated in the control of coronavirus subgenomic mRNA synthesis (transcription). By using comparative sequence analysis, we now show that the respective proteins of all sequenced coronaviruses are flanked by two conserved PLpro cleavage sites and share a complex (multi)domain organization with PL1pro being inactivated in IBV. Based upon these predictions, the processing of the human coronavirus 229E p195/p210 N terminus was studied in detail. First, an 87-kDa protein (p87), which is derived from a pp1a/pp1ab region immediately upstream of p195/p210, was identified in human coronavirus 229E infected cells. Second, in vitro synthesized proteins representing different parts of pp1a were autocatalytically processed at the predicted site. Surprisingly, both PL1pro and PL2pro cleaved between p87 and p195/p210. The PL1pro-mediated cleavage was slow and significantly suppressed by a non proteolytic activity of PL2pro. In contrast, PL2pro, whose proteolytic activity and specificity were established in this study, cleaved the same site efficiently in the presence of the upstream domains. Third, a correlation was observed between the overlapping substrate specificities and the parallel evolution of PL1pro and PL2pro. Collectively, our results imply that the p195/p210 autoprocessing mechanisms may be conserved among coronaviruses to an extent not appreciated previously, with PL2pro playing a major role. A large subset of coronaviruses may employ two proteases to cleave the same site(s) and thus regulate the expression of the viral genome in a unique way. PMID- 11431477 TI - Eukaryotic-like adenylyl cyclases in Mycobacterium tuberculosis H37Rv: cloning and characterization. AB - Screening the Mycobacterium tuberculosis H37Rv genomic library for complementation of catabolic defect for cAMP-dependent expression of maltose operon produced the adenylyl cyclase gene (Mtb cya, (1997)) annotated later as Rv1625c (Cole, S. T., Brosch, R., Parkhill, J., Garnier, T., Churcher, C., Harris, D., Gordon, S. V., Eiglmeier, K., Gas, S., Barry, C. E., III, et al. (1998) Nature 393, 537-544). The deduced amino acid (aa) sequence (443 aa) encoded by Mtb cya contains a single hydrophobic domain of six transmembrane helices (152 aa) in the amino-terminal half of the protein. Flanking this domain are an arginine-rich (17%) amino-terminal cytoplasmic tail (46 aa) and a carboxyl terminal cytoplasmic domain (245 aa) with extensive homology to the catalytic core of eukaryotic adenylyl cyclases. Site-directed mutagenesis of Arg(43) and Arg(44) to alanine/glycine showed a loss of adenylyl cyclase activity, whereas mutagenesis to lysine restored the activity. Hence it is proposed that the formation of the catalytic site in Mtb adenylyl cyclase requires an interaction between Arg(43) and Arg(44) residues in the distal cytoplasmic tail and the carboxyl-terminal cytoplasmic domain. Mtb adenylyl cyclase activity at the physiological concentration of ATP (1 mm) was 475 nmol of cAMP/min/mg of membrane protein in the presence of Mn(2+) but only 10 nmol of cAMP/min/mg of membrane protein in the presence of Mg(2+). The physiological significance of the activation of Mtb adenylyl cyclase by Mn(2+) is discussed in view of the presence of manganese transporter protein in mycobacteria and macrophages wherein mycobacteria reside. PMID- 11431478 TI - Integrins regulate the intracellular distribution of eukaryotic initiation factor 4E in platelets. A checkpoint for translational control. AB - Recent evidence from our laboratory demonstrates that platelets synthesize numerous proteins in a signal-dependent fashion (Pabla, R., Weyrich, A. S., Dixon, D. A., Bray, P. F., McIntyre, T. M., Prescott, S. M., and Zimmerman, G. A. (1999) J. Cell Biol. 144, 175-184; Weyrich, A. S., Dixon, D. A., Pabla, R., Elstad, M. R., McIntyre, T. M., Prescott, S. M., and Zimmerman, G. A. (1998) Proc. Natl. Acad. Sci. U. S. A. 95, 5556-5561). Protein synthesis in platelets is controlled at the translational level; however, the mechanisms of regulation are not known. Here we demonstrate that translation initiation factors are redistributed to mRNA-rich areas in aggregated platelets, an event that induces protein synthesis. Interrogation of cDNA arrays revealed that platelet-derived mRNAs are primarily associated with the cytoskeletal core. In contrast, eukaryotic initiation factor 4E (eIF4E), the essential mRNA cap-binding protein that controls global translation rates, is localized in the membrane skeleton and soluble fraction of platelets, physically separated from most mRNAs. Platelet activation redistributes eIF4E to the cytoskeleton and increases interactions of eIF4E with mRNA cap structures. Redistribution of eIF4E to the mRNA-rich cytoskeleton coincides with a marked increase in protein synthesis, a process that is blocked when intracellular actin is disrupted. Additional studies demonstrated that beta(3) integrins are the primary membrane receptor that distributes eIF4E within the cell. These results imply that integrins link receptor-mediated pathways with mRNA-rich cytoskeletal domains and thereby modulate the organization of intracellular translational complexes. They also indicate that the functional status of eIF4E is regulated by its intracellular distribution. PMID- 11431479 TI - A novel nuclear human poly(A) polymerase (PAP), PAP gamma. AB - Poly(A) polymerase (PAP) is present in multiple forms in mammalian cells and tissues. Here we show that the 90-kDa isoform is the product of the gene PAPOLG, which is distinct from the previously identified genes for poly(A) polymerases. The 90-kDa isoform is referred to as human PAP gamma (hsPAP gamma). hsPAP gamma shares 60% identity to human PAPII (hsPAPII) at the amino acid level. hsPAP gamma exhibits fundamental properties of a bona fide poly(A) polymerase, specificity for ATP, and cleavage and polyadenylation specificity factor/hexanucleotide dependent polyadenylation activity. The catalytic parameters indicate similar catalytic efficiency to that of hsPAPII. Mutational analysis and sequence comparison revealed that hsPAP gamma and hsPAPII have similar organization of structural and functional domains. hsPAP gamma contains a U1A protein-interacting region in its C terminus, and PAP gamma activity can be inhibited, as hsPAPII, by the U1A protein. hsPAPgamma is restricted to the nucleus as revealed by in situ staining and by transfection experiments. Based on this and previous studies, it is obvious that multiple isoforms of PAP are generated by three distinct mechanisms: gene duplication, alternative RNA processing, and post-translational modification. The exclusive nuclear localization of hsPAP gamma establishes that multiple forms of PAP are unevenly distributed in the cell, implying specialized roles for the various isoforms. PMID- 11431480 TI - Differential involvement of initiator caspases in apoptotic volume decrease and potassium efflux during Fas- and UV-induced cell death. AB - Caspase activation and apoptotic volume decrease are fundamental features of programmed cell death; however, the relationship between these components is not well understood. Here we provide biochemical and genetic evidence for the differential involvement of initiator caspases in the apoptotic volume decrease during both intrinsic and extrinsic activation of apoptosis. Apoptosis induction in Jurkat T lymphocytes by Fas receptor engagement (intrinsic) or ultraviolet (UV)-C radiation (extrinsic) triggered the loss of cell volume, which was restricted to cells with diminished intracellular K(+) ions. These characteristics kinetically coincided with the proteolytic processing and activation of both initiator and effector caspases. Although the polycaspase inhibitor benzyloxycarbonyl-Val-Ala-Asp fluoromethyl ketone completely inhibited the Fas-mediated apoptotic volume decrease and K(+) efflux, it was much less effective in preventing these processes during UV-induced cell death under conditions whereby caspase activities and DNA degradation were blocked. To define the roles of specific initiator caspases, we utilized Jurkat cells genetically deficient in caspase-8 or stably transfected with a dominant-negative mutant of caspase-9. The results show that the activation of caspase-8, but not caspase-9, is necessary for Fas-induced apoptosis. Conversely, caspase-9, but not caspase-8, is important for UV-mediated shrunken morphology and apoptosis progression. Together, these findings indicate that cell shrinkage and K(+) efflux during apoptosis are tightly coupled, but are differentially regulated by either caspase 8 or caspase-9 depending on specific pathways of cell death. PMID- 11431481 TI - G-protein-coupled receptor activation induces the membrane translocation and activation of phosphatidylinositol-4-phosphate 5-kinase I alpha by a Rac- and Rho dependent pathway. AB - Phosphatidylinositol 4,5-bisphosphate (PI4,5P(2)) mediates cell motility and changes in cell shape in response to extracellular stimuli. In platelets, it is synthesized from PI4P by PIP5K in response to stimulation of a G-protein-coupled receptor by an agonist, such as the thrombin. In the present study, we have addressed the pathway that induces PIP5K I alpha activation following the addition of thrombin. Under resting condition expressed PIP5K I alpha was predominantly localized in a perinuclear distribution. After stimulation of the thrombin receptor, PAR1, or overexpression of a constitutively active variant of G alpha(q), PIP5K I alpha translocated to the plasma membrane. Movement of PIP5K I alpha to the cell membrane was dependent on both GTP-bound Rac and Rho, but not Arf, because: 1) inactive GDP-bound variants of either Rac or Rho blocked the translocation induced by constitutively active G alpha(q), 2) constitutively GTP bound active variants of Rac or Rho induced PIP5K I alpha translocation in the absence of other stimuli, and 3) constitutively active variants of Arf1 or Arf6 failed to induce membrane translocation of PIP5K I alpha. In addition, a dominant negative variant of Rho blocked the PIP5K I alpha membrane translocation induced by constitutively active Rac, whereas dominant negative variants of either Rac or Arf6 failed to block PIP5K I alpha membrane translocation induced by constitutively active Rho. This implies that the effect on PIP5K I alpha by Rac is indirect, and requires the activation of Rho. In contrast to the findings with PIP5K I alpha, the related lipid kinase PIP4K failed to undergo translocation after stimulation by small GTP-binding proteins Rac or Rho. We also tested whether membrane localization of PIP5K I alpha correlated with an increase in its lipid kinase activity and found that co-expressing of PIP5K I alpha with either constitutively active G alpha(q), Rac, or Rho led to a 5- to 7-fold increase in PIP5K I alpha activity. Thus, these findings suggest that stimulation of a G protein-coupled receptor (PAR1) leads to the sequential activation of G alpha(q), Rac, Rho, and PIP5K I alpha. Once activated and translocated to the cell membrane, PIP5K I alpha becomes available to phosphorylate PI4P to generate PI4,5P(2) on the plasma membrane. PMID- 11431482 TI - Adiponutrin, a transmembrane protein corresponding to a novel dietary- and obesity-linked mRNA specifically expressed in the adipose lineage. AB - We have used a mRNA differential display technique to identify new genes involved in the reprogramming of gene expression during the adipose conversion of mouse 3T3 preadipocyte cell lines. We report here on the identification and cloning of a novel adipose-specific cDNA encoding a predicted membrane protein of 413 amino acids. The level of the corresponding 3.2-kilobase mRNA is tremendously increased during 3T3-L1 and 3T3-F442A differentiation into adipocytes. A single, very abundant 3.2-kilobase transcript is also found in inguinal and epididymal white adipose tissues and in interscapular brown adipose tissue but not in any other tissues examined. Its expression in adipose tissue is under tight nutritional regulation. The level of this novel 3.2-kilobase transcript becomes virtually nondetectable during fasting but is dramatically increased when fasted mice are refed a high carbohydrate diet. Based on its adipose specificity and dietary regulation, the novel gene product has been designated adiponutrin. The expression of adiponutrin mRNA is also 50-fold elevated in genetically obese fa/fa rats, indicating a link between adiponutrin and obesity. Western blot and confocal imagery analyses with epitope-tagged protein transiently expressed in 3T3-L1 adipocytes, and COS cells show that adiponutrin strictly localizes to membranes and is absent from the cytosol. Sequence analysis reveals homologies with several other members of related eukaryotic proteins, including a human paralog, which has been recently described in vesicular transport mechanisms. This leads us to suggest that adiponutrin could be involved in vesicular targeting and protein transport restricted to the adipocyte function. PMID- 11431483 TI - Molecular and Biochemical Characterization of Rat epsilon -N-Trimethyllysine Hydroxylase, the First Enzyme of Carnitine Biosynthesis. AB - epsilon-N-Trimethyllysine hydroxylase (EC ) is the first enzyme in the biosynthetic pathway of l-carnitine and catalyzes the formation of beta-hydroxy-N epsilon-trimethyllysine from epsilon-N-trimethyllysine, a reaction dependent on alpha-ketoglutarate, Fe(2+), and oxygen. We purified the enzyme from rat kidney and sequenced two internal peptides by quadrupole-time-of-flight mass spectroscopy. The peptide sequences were used to search the Expressed Sequence Tag data base, which led to the identification of a rat cDNA of 1218 base pairs encoding a polypeptide of 405 amino acids with a calculated molecular mass of 47.5 kDa. Using the rat sequence we also identified the homologous cDNAs from human and mouse. Heterologous expression of both the rat and human cDNAs in COS cells confirmed that they encode epsilon-N-trimethyllysine hydroxylase. Subcellular fractionation studies revealed that the rat enzyme is localized exclusively in mitochondria. Expression studies in yeast indicated that the rat enzyme is synthesized as a 47.5-kDa precursor and subsequently processed to a mature protein of 43 kDa, presumably upon import in mitochondria. The Michaelis Menten constants of the purified rat enzyme for trimethyllysine, alpha ketoglutarate, and Fe(2+) were 1.1 mm, 109 microm, and 54 microm, respectively. Both gel filtration and blue native polyacrylamide gel electrophoresis analysis showed that the native enzyme has a mass of approximately 87 kDa, indicating that in rat epsilon-N-trimethyllysine hydroxylase is a homodimer. PMID- 11431484 TI - Saccharomyces cerevisiae Adr1p governs fatty acid beta-oxidation and peroxisome proliferation by regulating POX1 and PEX11. AB - Saccharomyces cerevisiae Adr1p is essential for fatty acid degradation and peroxisome proliferation. Here, the role of Adr1p was examined with respect to the transcriptional regulation of the Pip2p-Oaf1p dependent genes POX1 and PEX11. POX1 encodes the rate-limiting enzyme of peroxisomal beta-oxidation, acyl-CoA oxidase. The POX1 promoter was shown to contain a canonical Adr1p element (UAS1), within which the oleate response element (ORE) was nested. PEX11 codes for a peroxin that is critical for normal peroxisome proliferation, and its promoter was shown similarly to contain a UAS1-like element overlapping the ORE. Northern analysis demonstrated that transcriptional up-regulation of both POX1 and PEX11 was abolished in adr1 Delta mutant cells, and immunoblotting confirmed that the abundance of their gene products was dramatically reduced. Studies of an overlapping ORE/UAS1 arrangement in the CTA1 promoter revealed synergy between these elements. We conclude that overlapping ORE and UAS1 elements in conjunction with their binding factors Pip2p-Oaf1p and Adr1p coordinate the carbon flux through beta-oxidation with peroxisome proliferation. PMID- 11431485 TI - Impact of network activities on neuronal properties in corticothalamic systems. AB - Data from in vivo and in vitro experiments are discussed to emphasize that synaptic activities in neocortex and thalamus have a decisive impact on intrinsic neuronal properties in intact-brain preparations under anesthesia and even more so during natural states of vigilance. Thus the firing patterns of cortical neuronal types are not inflexible but may change with the level of membrane potential and during periods rich in synaptic activity. The incidences of some cortical cell classes (defined by their responses to depolarizing current pulses) are different in isolated cortical slabs in vivo or in slices maintained in vitro compared with the intact cortex of naturally awake animals. Network activities, which include the actions of generalized modulatory systems, have a profound influence on the membrane potential, apparent input resistance, and backpropagation of action potentials. The analysis of various oscillatory types leads to the conclusion that in the intact brain, there are no "pure" rhythms, generated in simple circuits, but complex wave sequences (consisting of different, low- and fast-frequency oscillations) that result from synaptic interactions in corticocortical and corticothalamic neuronal loops under the control of activating systems arising in the brain stem core or forebrain structures. As an illustration, it is shown that the neocortex governs the synchronization of network or intrinsically generated oscillations in the thalamus. The rhythmic recurrence of spike bursts and spike trains fired by thalamic and cortical neurons during states of decreased vigilance may lead to plasticity processes in neocortical neurons. If these phenomena, which may contribute to the consolidation of memory traces, are not constrained by inhibitory processes, they induce seizures in which the neocortex initiates the paroxysms and controls their thalamic reflection. The results indicate that intact-brain preparations are necessary to investigate global brain functions such as behavioral states of vigilance and paroxysmal activities. PMID- 11431486 TI - Cannabinoid-induced presynaptic inhibition of glutamatergic EPSCs in substantia gelatinosa neurons of the rat spinal cord. AB - The effect of cannabinoids on excitatory transmission in the substantia gelatinosa was investigated using intracellular recording from visually identified neurons in a transverse slice preparation of the juvenile rat spinal cord. In the presence of strychnine and bicuculline, perfusion of the cannabinoid receptor agonist WIN55,212-2 reduced the frequency and the amplitude of spontaneous excitatory postsynaptic currents (sEPSCs). Furthermore, the frequency of miniature EPSCs (mEPSCs) was also decreased by WIN55,212-2, whereas their amplitude was not affected. Similar effects were reproduced using the endogenous cannabinoid ligand anandamide. The effects of both agonists were blocked by the selective CB(1) receptor antagonist SR141716A. Electrical stimulation of high threshold fibers in the dorsal root evoked a monosynaptic EPSC in lamina II neurons. In the presence of WIN55,212-2, the amplitude of the evoked EPSC (eEPSCs) was reduced, and the paired-pulse ratio was increased. The reduction of the eEPSC following CB(1) receptor activation was unlikely to have a postsynaptic origin because the response to AMPA, in the presence of 1 microM TTX, was unchanged. To investigate the specificity of this synaptic inhibition, we selectively activated the nociceptive C fibers with capsaicin, which induced a strong increase in the frequency of EPSCs. In the presence of WIN55,212-2, the response to capsaicin was diminished. In conclusion, these results strongly suggest a presynaptic location for CB(1) receptors whose activation results in inhibition of glutamate release in the spinal dorsal horn. The strong inhibitory effect of cannabinoids on C fibers may thereby contribute to the modulation of the spinal excitatory transmission, thus producing analgesia at the spinal level. PMID- 11431487 TI - GABA(A) and 5-HT(3) receptors are involved in dorsal root reflexes: possible role in periaqueductal gray descending inhibition. AB - The dorsal root reflex (DRR) is a measure of the central excitability of presynaptic inhibitory circuits in the spinal cord. Activation of the periaqueductal gray (PAG), a center for descending inhibition of spinal cord nociceptive transmission, induces release of variety of neurotransmitters in the spinal cord, including GABA and serotonin (5-HT). GABA has been shown to be involved in generation of DRRs. In this study, pharmacological agents that influence DRRs and their possible mechanisms were investigated. DRRs were recorded in anesthetized rats from filaments teased from the cut central stump of the left L(4) or L(5) dorsal root, using a monopolar recording electrode. Stimulating electrodes were placed either on the left sciatic nerve or transcutaneously in the left foot. Animals were paralyzed and maintained by artificial ventilation. Drugs were applied topically to the spinal cord. A total of 64 units were recorded in 34 Sprague-Dawley rats. Peripheral receptive fields were found for nine of these units. In these units, DRRs were evoked by brush, pressure, and pinch stimuli. Nine units were tested for an effect of electrical stimulation in the periaqueductal gray on the DRRs. In eight cases, DRR responses were enhanced following PAG stimulation. The background activity was 4.2 +/- 1.9 spikes/s (mean +/- SE; range: 0-97.7; n = 57). The responses to agents applied to the spinal cord were (in spikes/s): artificial cerebrospinal fluid, 7.1 +/- 3.6 (range: 0-86.9; n = 25); 0.1 mM GABA, 16.8 +/- 8.7 (range: 0-191.0; n = 22); 1.0 mM GABA, 116.0 +/- 26.5 (range: 0.05-1001.2; n = 50); and 1.0 mM phenylbiguanide (PBG), 68.1 +/- 25.3 (range: 0-1,073.0; n = 49). Bicuculline (0.5 mM, n = 27) and ondansetron (1.0 mM, n = 10) blocked the GABA and PBG effects, respectively (P < 0.05). Significant cross blockade was also observed. It is concluded that GABA(A) receptors are likely to play a key role in the generation of DRRs, but that 5 HT(3) receptors may also contribute. DRRs can be modulated by supraspinal mechanisms through descending systems. PMID- 11431488 TI - Models of respiratory rhythm generation in the pre-Botzinger complex. III. Experimental tests of model predictions. AB - We used the testable predictions of mathematical models proposed by Butera et al. to evaluate cellular, synaptic, and population-level components of the hypothesis that respiratory rhythm in mammals is generated in vitro in the pre-Botzinger complex (pre-BotC) by a heterogeneous population of pacemaker neurons coupled by fast excitatory synapses. We prepared thin brain stem slices from neonatal rats that capture the pre-BotC and maintain inspiratory-related motor activity in vitro. We recorded pacemaker neurons extracellularly and found: intrinsic bursting behavior that did not depend on Ca(2+) currents and persisted after blocking synaptic transmission; multistate behavior with transitions from quiescence to bursting and tonic spiking states as cellular excitability was increased via extracellular K(+) concentration ([K(+)](o)); a monotonic increase in burst frequency and decrease in burst duration with increasing [K(+)](o); heterogeneity among different cells sampled; and an increase in inspiratory burst duration and decrease in burst frequency by excitatory synaptic coupling in the respiratory network. These data affirm the basis for the network model, which is composed of heterogeneous pacemaker cells having a voltage-dependent burst generating mechanism dominated by persistent Na(+) current (I(NaP)) and excitatory synaptic coupling that synchronizes cell activity. We investigated population-level activity in the pre-BotC using local "macropatch" recordings and confirmed these model predictions: pre-BotC activity preceded respiratory-related motor output by 100-400 ms, consistent with a heterogeneous pacemaker-cell population generating inspiratory rhythm in the pre-BotC; pre-BotC population burst amplitude decreased monotonically with increasing [K(+)](o) (while frequency increased), which can be attributed to pacemaker cell properties; and burst amplitude fluctuated from cycle to cycle after decreasing bilateral synaptic coupling surgically as predicted from stability analyses of the model. We conclude that the pacemaker cell and network models explain features of inspiratory rhythm generation in vitro. PMID- 11431489 TI - Activation of GABA(A) receptors in subthalamic neurons in vitro: properties of native receptors and inhibition mechanisms. AB - The subthalamic nucleus (STN) influences the output of the basal ganglia, thereby interfering with motor behavior. The main inputs to the STN are GABAergic. We characterized the GABA(A) receptors expressed in the STN and investigated the response of subthalamic neurons to the activation of GABA(A) receptors. Cell attached and whole cell recordings were made from rat brain slices using the patch-clamp technique. The newly identified epsilon subunit confers atypical pharmacological properties on recombinant receptors, which are insensitive to barbiturates and benzodiazepines. We tested the hypothesis that native subthalamic GABA(A) receptors contain epsilon proteins. Applications of increasing concentrations of muscimol, a selective GABA(A) agonist, induced Cl(-) and HCO currents with an EC(50) of 5 microM. Currents induced by muscimol were fully blocked by the GABA(A) receptor antagonists, bicuculline and picrotoxin. They were strongly potentiated by the barbiturate, pentobarbital (+190%), and by the benzodiazepines, diazepam (+197%) and flunitrazepam (+199%). Spontaneous inhibitory postsynaptic currents were also significantly enhanced by flunitrazepam. Furthermore, immunohistological experiments with an epsilon subunit-specific antibody showed that the epsilon protein was not expressed within the STN. Native subthalamic GABA(A) receptors did not, therefore, display pharmacological or structural properties consistent with receptors comprising epsilon. Burst firing is a hallmark of Parkinson's disease. Half of the subthalamic neurons have the intrinsic capacity of switching from regular-firing to burst-firing mode when hyperpolarized by current injection. This raises the possibility that activation of GABA(A) receptors might trigger the switch. Statistical analysis of spiking activity established that 90% of intact neurons in vitro were in single-spike firing mode, whereas 10% were in burst-firing mode. Muscimol reversibly stopped recurrent electrical activity in all intact neurons. In neurons held in whole cell configuration, membrane potential hyperpolarized by -10 mV whilst input resistance decreased by 50%, indicating powerful membrane shunting. Muscimol never induced burst firing, even in neurons that exhibited the capacity of switching from regular- to burst-firing mode. These molecular and functional data indicate that native subthalamic GABA(A) receptors do not contain the epsilon protein and activation of GABA(A) receptors induces membrane shunting, which is essential for firing inhibition but prevents switching to burst-firing. They suggest that the STN, like many other parts of the brain, has the physiological and structural features of the widely expressed GABA(A) receptors consisting of alphabetagamma subunits. PMID- 11431490 TI - Long-term recordings of networks of immortalized GnRH neurons reveal episodic patterns of electrical activity. AB - The CNS controls reproduction through pulsatile secretion of gonadotropin releasing hormone (GnRH). Episodic increases in the firing rate of unidentified hypothalamic neurons have been associated with downstream markers of GnRH secretion. Whether this episodic electrical activity is intrinsic to GnRH neurons, intrinsic to other "pulse generator" neurons that drive GnRH neurons, or a combination of these is unknown. To determine if GnRH neurons display episodic firing patterns in isolation from other cell types, immortalized GnRH neurons (GT1-7 cells) were cultured on multiple microelectrode arrays. Long-term, multi site recordings of GT1-7 cells revealed repeated episodes of increased firing rate with an interval of 24.8 +/- 1.3 (SE) min that were completely eliminated by tetrodotoxin, a sodium channel blocker. This pattern was comprised of active units that fired independently as well as coincidentally, suggesting the overall pattern of electrical activity in GT1-7 cells emerges as a network property. The A-type potassium-channel antagonist 4-aminopyridine (1 mM) increased both firing rate and GnRH secretion, demonstrating the presence of A-type currents in these cells and supporting the hypothesis that electrical activity is associated with GnRH release. Physiologically relevant episodic firing patterns are thus an intrinsic property of immortalized GnRH neurons and appear to be associated with secretion. The finding that overall activity is derived from the sum of multiple independent active units within a network may have important implications for the genesis of the GnRH secretory pattern that is delivered to the target organ. Specifically, these data suggest not every GnRH neuron participates in each secretory pulse and provide a possible mechanism for the variations in GnRH-pulse amplitude observed in vivo. PMID- 11431491 TI - Role of inhibition in cortical reorganization of the adult raccoon revealed by microiontophoretic blockade of GABA(A) receptors. AB - Cortical reorganization was induced by amputation of the 4th digit in 11 adult raccoons. Animals were studied at various intervals, ranging from 2 to 37 wk, after amputation. Recordings were made from a total of 129 neurons in the deafferented cortical region using multibarrel micropipettes. Several types of receptive fields were described in reorganized cortex: restricted fields were similar in size to the normal receptive fields in nonamputated animals; multi regional fields included sensitive regions on both adjacent digits and/or the underlying palm and were either continuous over the entire field or consisted of split fields. The proportion of neurons with restricted fields increased with time after amputation and was greater than previously found in subcortical regions. A GABA(A) receptor antagonist (bicuculline methiodide), glutamate, and GABA were administered iontophoretically to these neurons while determining their receptive fields and thresholds. Bicuculline administration resulted in expansion of the receptive field in 60% of the 93 neurons with cutaneous fields. In most cases (33 neurons) this consisted of a simple expansion around the borders of the predrug receptive field, and the average expansion (426%) was not different from that seen in nonamputated animals. In some neurons (n = 4), bicuculline produced an expansion from one digit onto the adjacent palm or another digit, an effect never seen in control animals. Bicuculline also changed the split fields of seven neurons into continuous fields by exposing a responsive region between the split fields. Finally, bicuculline changed the internal receptive field organization of 10 neurons by revealing subfields with reduced thresholds. In contrast to the situation in nonamputated animals, iontophoretic administration of glutamate also produced receptive field expansion in some neurons (n = 6), but the size and/or shape of the change was different from that produced by bicuculline, indicating that the effects of bicuculline were not due to an overall facilitation of neuronal activity. These results are consistent with the hypotheses that an important component of long-term cortical reorganization is the gradual reduction in effective receptive field size and that intracortical inhibitory networks are partially responsible for these changes. PMID- 11431492 TI - Identification of two types of inspiratory pacemaker neurons in the isolated respiratory neural network of mice. AB - In the respiratory network of mice, we characterized with the whole cell patch clamp technique pacemaker properties in neurons discharging in phase with inspiration. The respiratory network was isolated in a transverse brain stem slice containing the pre-Botzinger complex (PBC), the presumed site for respiratory rhythm generation. After blockade of respiratory network activity with 6-cyano-7-nitroquinoxalene-2,3-dione (CNQX), 18 of 52 inspiratory neurons exhibited endogenous pacemaker activity, which was voltage dependent, could be reset by brief current injections and could be entrained by repetitive stimuli. In the pacemaker group (n = 18), eight neurons generated brief bursts (0.43 +/- 0.03 s) at a relatively high frequency (1.05 +/- 0.12 Hz) in CNQX. These bursts resembled the bursts that these neurons generated in the intact network during the interval between two inspiratory bursts. Cadmium (200 microM) altered but did not eliminate this bursting activity, while 0.5 microM tetrodotoxin suppressed bursting activity. Another set of pacemaker neurons (10 of 18) generated in CNQX longer bursts (1.57 +/- 0.07 s) at a lower frequency (0.35 +/- 0.01 Hz). These bursts resembled the inspiratory bursts generated in the intact network in phase with the population activity. This bursting activity was blocked by 50-100 microM cadmium or 0.5 microM tetrodotoxin. We conclude that the respiratory neural network contains pacemaker neurons with two types of bursting properties. The two types of pacemaker activities might have different functions within the respiratory network. PMID- 11431493 TI - Three-dimensional binocular kinematics of torsional vestibular nystagmus during convergence on head-fixed targets in humans. AB - When a human subject is oscillated about the nasooccipital axis and fixes upon targets along the horizontal head-fixed meridian, angular eye velocity includes a vertical component that increases with the horizontal eccentricity of the line-of sight. This vertical eye movement component is necessary to prevent retinal slip. We asked whether fixation on a near head-fixed target during the same torsional vestibular stimulation would lead to differences of vertical eye movements between the right and the left eye, as the directions of the two lines-of-sight are not parallel during convergence. Healthy human subjects (n = 6) were oscillated (0.3 Hz, +/-30 degrees) about the nasooccipital axis on a three dimensional motor-driven turntable. Binocular movements were recorded using the dual search coil technique. A head-fixed laser dot was presented 1.4 m (far head fixed target) or 0.25 m (near head-fixed target) in front of the right eye. We found highly significant (P < 0.01) correlations (R binocular = 0.8, monocular = 0.59) between the convergence angle and the difference of the vertical eye velocity between the two eyes. The slope of the fitted linear regression between the two parameters (s = 0.45) was close to the theoretical slope necessary to prevent vertical retinal slippage (predicted s = 0.5). Covering the left eye did not significantly change the slope (s = 0.52). In addition, there was a marked gain reduction (approximately 35%) of the torsional vestibuloocular reflex (VOR) between viewing the far and the near targets, confirming earlier results by others. There was no difference in torsional gain reduction between the two eyes. Lenses of +3 dpt positioned in front of both eyes to decrease the amount of accommodation did not further change the gain of the torsional VOR. In conclusion, ocular convergence on a near head-fixed target during torsional vestibular stimulation leads to deviations in vertical angular velocity between the two eyes necessary to prevent vertical double vision. The vertical deviation velocity is mainly linked to the amount of convergence, since it also occurs during monocular viewing of the near head-fixed target. This suggests that convergence during vestibular stimulation automatically leads to an alignment of binocular rotation axes with the visual axes independent of retinal slip. PMID- 11431494 TI - Removal of NMDA receptor Mg(2+) block extends the action of NT-3 on synaptic transmission in neonatal rat motoneurons. AB - NT-3 has previously been reported to enhance AMPA/kainate receptor-mediated synaptic responses in motoneurons via an effect on the N-methyl-D-aspartate (NMDA) receptor. To investigate neurotrophin-3 (NT-3) action further, we measured the NMDA receptor (NMDAR)-mediated synaptic response directly by intracellular recording in motoneurons after blocking AMPA/kainate, GABA(A), GABA(B) and glycine receptor-mediated responses pharmacologically. Two pathways were stimulated, the segmental dorsal root (DR) and the descending ventrolateral fasciculus (VLF). The DR-evoked NMDAR-mediated response in motoneurons of rats younger than 1 wk has two components, the initial one of which is generated monosynaptically. NT-3 strongly potentiated both NMDA components in a rapidly reversible manner. No NMDAR-mediated responses were present at VLF connections and at DR connections in older (1- to 2-wk-old) neonates. Bath-applied NT-3 induced potentiation of the AMPA/kainate receptor-mediated response occurred only at connections that exhibit a synaptic NMDA receptor-mediated response. Reducing Mg(2+) concentration in the bathing solution restored the NMDAR-mediated response elicited by DR stimulation in older neonates and by VLF throughout the neonatal period (0-2 wk). In low-Mg(2+), NT-3 enhanced AMPA/kainate receptor-mediated responses elicited by inputs normally not influenced by NT-3. Thus a major reason for the loss of NT-3 action on AMPA/kainate synaptic responses is the reduced activity of the NMDA receptor due to developing Mg(2+) block of NMDA receptor channel complex as the animal matures, and both can be re-established by reducing Mg(2+) concentration in fluid bathing the spinal cord. PMID- 11431495 TI - KT3.2 and KT3.3, two novel human two-pore K(+) channels closely related to TASK 1. AB - We report the cloning of human KT3.2 and KT3.3 new members of the two-pore K(+) channel (KT) family. Based on amino acid sequence and phylogenetic analysis, KT3.2, KT3.3, and TASK-1 constitute a subfamily within the KT channel mammalian family. When Xenopus oocytes were injected with KT3.2 cRNA, the resting membrane potential was brought close to the potassium equilibrium potential. At low extracellular K(+) concentrations, two-electrode voltage-clamp recordings revealed the expression of predominantly outward currents. With high extracellular K(+) (98 mM), the current-voltage relationship exhibited weak outward rectification. Measurement of reversal potentials at different [K(+)](o) revealed a slope of 48 mV per 10-fold change in K(+) concentration as expected for a K(+)-selective channel. Unlike TASK-1, which is highly sensitive to changes of pH in the physiological range, KT3.2 currents were relatively insensitive to changes in intracellular or extracellular pH within this range due to a shift in the pH dependency of KT3.2 of 1 pH unit in the acidic direction. On the other hand, the phorbol ester phorbol 12-myristate 13-acetate (PMA), which does not affect TASK-1, produces strong inhibition of KT3.2 currents. Human KT3.2 mRNA expression was most prevalent in the cerebellum. In rat, KT3.2 is exclusively expressed in the brain, but it has a wide distribution within this organ. High levels of expression were found in the cerebellum, medulla, and thalamic nuclei. The hippocampus has a nonhomogeneous distribution, expressing at highest levels in the lateral posterior and inferior portions. Medium expression levels were found in neocortex. The KT3.2 gene is located at chromosome 8q24 1-3, and the KT3.3 gene maps to chromosome 20q13.1. PMID- 11431496 TI - Modeling V1 disparity tuning to time-varying stimuli. AB - Most models of disparity selectivity consider only the spatial properties of binocular cells. However, the temporal response is an integral component of real neurons' activities, and time-varying stimuli are often used in the experiments of disparity tuning. To understand the temporal dimension of V1 disparity representation, we incorporate a specific temporal response function into the disparity energy model and demonstrate that the binocular interaction of complex cells is separable into a Gabor disparity function and a positive time function. We then investigate how the model simple and complex cells respond to widely used time-varying stimuli, including motion-in-depth patterns, drifting gratings, moving bars, moving random-dot stereograms, and dynamic random-dot stereograms. It is found that both model simple and complex cells show more reliable disparity tuning to time-varying stimuli than to static stimuli, but similarities in the disparity tuning between simple and complex cells depend on the stimulus. Specifically, the disparity tuning curves of the two cell types are similar to each other for either drifting sinusoidal gratings or moving bars. In contrast, when the stimuli are dynamic random-dot stereograms, the disparity tuning of simple cells is highly variable, whereas the tuning of complex cells remains reliable. Moreover, cells with similar motion preferences in the two eyes cannot be truly tuned to motion in depth regardless of the stimulus types. These simulation results are consistent with a large body of extant physiological data, and provide some specific, testable predictions. PMID- 11431497 TI - Hypoxia-induced respiratory patterned activity in Lymnaea originates at the periphery. AB - Respiration in Lymnaea is a hypoxia-driven rhythmic behavior, which is controlled by an identified network of central pattern generating (CPG) neurons. However, the precise site(s) (i.e., central or peripheral) at which hypoxia acts and the cellular mechanisms by which the respiratory chemosensory drive is conveyed to the CPG were previously unknown. Using semi-intact and isolated ganglionic preparations, we provide the first direct evidence that the hypoxia-induced respiratory drive originates at the periphery (not within the central ring ganglia) and that it is conveyed to the CPG neurons via the right pedal dorsal neuron 1 (RPeD1). The respiratory discharge frequency increased when the periphery, but not the CNS, was made hypoxic. We found that in the semi-intact preparations, the frequency of spontaneously occurring respiratory bursts was significantly lower than in isolated ganglionic preparations. Thus the periphery exerts a suppressive regulatory control on respiratory discharges in the intact animal. Moreover, both anoxia (0% O(2)) and hypercapnia (10% CO(2)) produce a reduction in respiratory discharges in semi-intact, but not isolated preparations. However, the effects of CO(2) may be mediated through pH changes of the perfusate. Finally, we demonstrate that chronic exposure of the animals to hypoxia (90% N(2)), prior to intracellular recordings, significantly enhanced the rate of spontaneously occurring respiratory discharges in semi-intact preparations, even if they were maintained in normoxic saline for several hours. Moreover, we demonstrate that the peripherally originated hypoxia signal is likely conveyed to the CPG neurons via RPeD1. In summary, the data presented in this study demonstrate the important role played by the periphery and the RPeD1 neuron in regulating respiration in response to hypoxia in Lymnaea. PMID- 11431498 TI - Network activity evoked by neocortical stimulation in area 36 of the guinea pig perirhinal cortex. AB - The perirhinal cortex is a key structure involved in memory consolidation and retrieval. In spite of the extensive anatomical studies that describe the intrinsic and extrinsic associative connections of the perirhinal cortex, the activity generated within such a network has been poorly investigated. We describe here the pattern of synaptic interactions that subtend the responses evoked in area 36 of the perirhinal cortex by neocortical and local stimulation. The experiments were carried out in the in vitro isolated guinea pig brain. The synaptic perirhinal circuit was reconstructed by integrating results obtained during intracellular recordings from layer II-III neurons with simultaneous current source density analysis of laminar profiles performed with 16-channel silicon probes. Both neocortical and local stimulation of area 36 determined a brief monosynaptic excitatory potential in layer II-III neurons, followed by a biphasic synaptic inhibitory potential possibly mediated by a feed-forward inhibitory circuit at sites close to the stimulation electrode and a late excitatory postsynaptic potential (EPSP) that propagated at distance within area 36 along the rhinal sulcus. During a paired-pulse stimulation test, the inhibitory postsynaptic potential (IPSP) and the late EPSP were abolished in the second conditioned response, suggesting that they are generated by poli-synaptic circuits. Current source density analysis of the field responses demonstrated that 1) the monosynaptic activity was generated in layers II-III and 2) the sink associated to the disynaptic responses was localized within the superficial layer of area 36. We conclude that the neocortical input induces a brief monosynaptic excitation in area 36 of the perirhinal cortex, that is curtailed by a prominent inhibition and generates a recurrent excitatory associative response that travels at distance within area 36 itself. The results suggest that the perirhinal cortex network has the potentials to integrate multimodal incoming neocortical information on its way to the hippocampus. PMID- 11431499 TI - Gadolinium reduces AMPA receptor desensitization and deactivation in hippocampal neurons. AB - The actions of the trivalent cation Gd(3+) on whole cell AMPA receptor-mediated currents were studied in isolated hippocampal neurons, in nucleated or outside out patches taken from cultured hippocampal neurons, and on miniature excitatory postsynaptic currents (mEPSCs) recorded in cultured hippocampal neurons. Glutamate, AMPA, or kainate was employed to activate AMPA receptors. Applications of relatively low concentrations of Gd(3+) (0.1-10 microM) substantially enhanced steady-state whole cell glutamate and kainate-evoked currents without altering peak currents, suggesting that desensitization was reduced. However, higher concentrations (>30 microM) depressed steady-state currents, indicating an underlying inhibition of channel activity. Lower concentrations of Gd(3+) also increased the potency of peak glutamate-evoked currents without altering that of steady-state currents. An ultrafast perfusion system and nucleated patches were then used to better resolve peak glutamate-evoked currents. Low concentrations of Gd(3+) reduced peak currents, enhanced steady-state currents, and slowed the onset of desensitization, providing further evidence that this cation reduces desensitization. In the presence of cyclothiazide, a compound that blocks desensitization, a low concentration Gd(3+) inhibited both peak and steady-state currents, indicating that Gd(3+) both reduces desensitization and inhibits these currents. Gd(3+) reduced the probability of channel opening at the peak of the currents but did not alter the single channel conductance calculated using nonstationary variance analysis. Recovery from desensitization was enhanced, and glutamate-evoked current activation and deactivation were slowed by Gd(3+). The Gd(3+)-induced reduction in desensitization did not require the presence of the GluR2 subunit as this effect was seen in hippocampal neurons from GluR2 null mutant mice. Gd(3+) reduced the time course of decay of mEPSCs perhaps as a consequence of its slowing of AMPA receptor deactivation although an increase in the frequency of mEPSCs also suggested enhanced presynaptic release of transmitter. These results demonstrate that Gd(3+) potently reduces AMPA receptor desensitization and mimics a number of the properties of the positive modulators of AMPA receptor desensitization such as cyclothiazide. PMID- 11431500 TI - Effects of bis(7)-tacrine on spontaneous synaptic activity and on the nicotinic ACh receptor of Torpedo electric organ. AB - Bis(7)-tacrine is a potent acetylcholinesterase inhibitor in which two tacrine molecules are linked by a heptylene chain. We tested the effects of bis(7) tacrine on the spontaneous synaptic activity. Miniature endplate potentials (MEPPs) were recorded extracellularly on slices of electric organ of Torpedo marmorata. Bis(7)-tacrine, at a concentration of 100 nM, increased the magnitudes that describe MEPPs: amplitude, area, rise time, rate of rise, and half-width. We also tested the effect of bis(7)-tacrine on nicotinic acetylcholine receptors by analyzing the currents elicited by acetylcholine (100 microM) in Torpedo electric organ membranes transplanted in Xenopus laevis oocytes. Bis(7)-tacrine inhibited the acetylcholine-induced currents in a reversible manner (IC(50) = 162 nM). The inhibition of nicotinic acetylcholine receptors was not voltage dependent, and bis(7)-tacrine increased the desensitization of nicotinic acetylcholine receptors. The Hill coefficient for bis(7)-tacrine was -0.72 +/- 0.02, indicating that bis(7)-tacrine binds to the nicotinic acetylcholine receptor in a molecular ratio of 1:1, but does not affect the binding of alpha-bungarotoxin with the nicotinic acetylcholine receptor. In conclusion, bis(7)-tacrine greatly increases the spontaneous quantal release from peripheral cholinergic terminals at a much lower concentration than tacrine. Bis(7)-tacrine also blocks acetylcholine induced currents of Torpedo electric organ, although the mechanism is different from that of tacrine: bis(7)-tacrine enhances desensitization, whereas tacrine reduces it. PMID- 11431501 TI - A novel Ca(2+) influx pathway in mammalian primary sensory neurons is activated by caffeine. AB - Single-cell microfluorimetry and electrophysiology techniques were used to identify and characterize a novel Ca(2+) influx pathway in adult rabbit vagal sensory neurons. Acutely dissociated nodose ganglion neurons (NGNs) exhibit robust Ca(2+)-induced Ca(2+) release (CICR) that can be triggered by 10 mM caffeine, the classic agonist of CICR. A caffeine-induced increase in cytosolic free Ca(2+) concentration ([Ca(2+)](i)) is considered diagnostic evidence of the existence of CICR. However, when CICR was disabled through depletion of intracellular Ca(2+) stores or pharmacological blockade of intracellular Ca(2+) release channels (ryanodine receptors), caffeine still elicited a significant rise in [Ca(2+)](i) in approximately 50% of NGNs. The same response was not elicited by pharmacological agents that elevate cyclic nucleotide concentrations. Moreover, extracellular Ca(2+) was obligatory for such caffeine-induced [Ca(2+)](i) rises in this population of NGNs, suggesting that Ca(2+) influx is responsible for this rise. Simultaneous microfluorimetry with whole cell patch clamp studies showed that caffeine activates an inward current that temporally parallels the rise in [Ca(2+)](i). The inward current had a reversal potential of +8.1 +/- 6.1 (SE) mV (n = 4), a mean peak amplitude of -126 +/- 24 pA (n = 4) at E(m) = -50 mV, and a slope conductance of 1.43 +/- 0.79 nS (n = 4). Estimated EC(50) values for caffeine-induced CICR and for caffeine-activated current were 1.5 and approximately 0.6 mM, respectively. These results indicate that caffeine induced rises in [Ca(2+)](i), in the presence of extracellular Ca(2+), can no longer be interpreted as unequivocal diagnostic evidence for CICR in neurons. These results also indicate that sensory neurons possess a novel Ca(2+) influx pathway. PMID- 11431502 TI - Synaptic drive to motoneurons during fictive swimming in the developing zebrafish. AB - The development of swimming behavior and the correlated activity patterns recorded in motoneurons during fictive swimming in paralyzed zebrafish larvae were examined and compared. Larvae were studied from when they hatch (after 2 days) and are first capable of locomotion to when they are active swimmers capable of capturing prey (after 4 days). High-speed (500 Hz) video imaging was used to make a basic behavioral characterization of swimming. At hatching and up to day 3, the larvae swam infrequently and in an undirected fashion. They displayed sustained bursts of contractions ('burst swimming') at an average frequency of 60-70 Hz that lasted from several seconds to a minute in duration. By day 4 the swimming had matured to a more frequent and less erratic "beat-and glide" mode, with slower (approximately 35 Hz) beats of contractions for approximately 200 ms alternating with glides that were twice as long, lasting from just a few cycles to several minutes overall. In whole cell current-clamp recordings, motoneurons displayed similar excitatory synaptic activity and firing patterns, corresponding to either fictive burst swimming (day 2-3) or beat-and glide swimming (day 4). The resting potentials were similar at all stages (about 70 mV) and the motoneurons were depolarized (to about -40 mV) with generally non overshooting action potentials during fictive swimming. The frequency of sustained inputs during fictive burst swimming and of repetitive inputs during fictive beat-and glide swimming corresponded to the behavioral contraction patterns. Fictive swimming activity patterns were eliminated by application of glutamate antagonists (kynurenic acid or 6-cyano-7-nitroquinoxalene-2,3-dione and DL-2-amino-5-phosphonovaleric acid) and were modified but maintained in the presence of the glycinergic antagonist strychnine. The corresponding synaptic currents underlying the synaptic drive to motoneurons during fictive swimming could be isolated under voltage clamp and consisted of cationic [glutamatergic postsynaptic currents (PSCs)] and anionic inputs (glycinergic PSCs). Either sustained or interrupted patterns of PSCs were observed during fictive burst or beat-and-glide swimming, respectively. During beat-and-glide swimming, a tonic inward current and rhythmic glutamatergic PSCs (approximately 35 Hz) were observed. In contrast, bursts of glycinergic PSCs occurred at a higher frequency, resulting in a more tonic pattern with little evidence for synchronized activity. We conclude that a rhythmic glutamatergic synaptic drive underlies swimming and that a tonic, shunting glycinergic input acts to more closely match the membrane time constant to the fast synaptic drive. PMID- 11431503 TI - Effects of acetylcholine and atropine on plasticity of central auditory neurons caused by conditioning in bats. AB - In the big brown bat (Eptesicus fuscus), conditioning with acoustic stimuli followed by electric leg-stimulation causes shifts in frequency-tuning curves and best frequencies (hereafter BF shifts) of collicular and cortical neurons, i.e., reorganization of the cochleotopic (frequency) maps in the inferior colliculus (IC) and auditory cortex (AC). The collicular BF shift recovers 180 min after the conditioning, but the cortical BF shift lasts longer than 26 h. The collicular BF shift is not caused by conditioning, as the AC is inactivated during conditioning. Therefore it has been concluded that the collicular BF shift is caused by the corticofugal auditory system. The collicular and cortical BF shifts both are not caused by conditioning as the somatosensory cortex is inactivated during conditioning. Therefore it has been hypothesized that the cortical BF shift is mostly caused by both the subcortical (e.g., collicular) BF shift and the activity of nonauditory systems such as the somatosensory cortex excited by an unconditioned leg-stimulation and the cholinergic basal forebrain. The main aims of our present studies are to examine whether acetylcholine (ACh) applied to the AC augments the collicular and cortical BF shifts caused by the conditioning and whether atropine applied to the AC abolishes the cortical BF shift but not the collicular BF shift, as expected from the preceding hypothesis. In the awake bat, we made the following findings. ACh applied to the AC augments not only the cortical BF shift but also the collicular BF shift through the corticofugal system. Atropine applied to the AC reduces the collicular BF shift and abolishes the cortical BF shift which otherwise would be caused. ACh applied to the IC significantly augments the collicular BF shift but affects the cortical BF shift only slightly. ACh makes the cortical BF shift long-lasting beyond 4 h, but it cannot make the collicular BF shift long-lasting beyond 3 h. Atropine applied to the IC abolishes the collicular BF shift. It reduces the cortical BF shift but does not abolish it. Our findings favor the hypothesis that the BF shifts evoked by the corticofugal system, and an increased ACh level in the AC evoked by the basal forebrain are both necessary to evoke a long-lasting cortical BF shift. PMID- 11431504 TI - Sensitivity of auditory cortical neurons to locations of signals and competing noise sources. AB - The present study examined cortical parallels to psychophysical signal detection and sound localization in the presence of background noise. The activity of single units or of small clusters of units was recorded in cortical area A2 of chloralose-anesthetized cats. Signals were 80-ms click trains that varied in location in the horizontal plane around the animal. Maskers were continuous broadband noises. In the focal masker condition, a single masker source was tested at various azimuths. In the diffuse masker condition, uncorrelated noise was presented from two speakers at +/-90 degrees lateral to the animal. For about 2/3 of units ("type A"), the presence of the masker generally reduced neural sensitivity to signals, and the effects of the masker depended on the relative locations of signal and masker sources. For the remaining 1/3 of units ("type B"), the masker reduced spike rates at low signal levels but often augmented spike rates at higher signal levels. Increases in spike rates of type B units were most common for signal sources in front of the ear contralateral to the recording site but tended to be independent of masker source location. For type A units, masker effects could be modeled as a shift toward higher levels of spike rate- and spike-latency-versus-level functions. For a focal masker, the shift size decreased with increasing separation of signal and masker. That result resembled psychophysical spatial unmasking, i.e., improved signal detection by spatial separation of the signal from the noise source. For the diffuse masker condition, the shift size generally was constant across signal locations. For type A units, we examined the effects of maskers on cortical signaling of sound source location, using an artificial-neural-network (ANN) algorithm. First, an ANN was trained to estimate the signal location in the quiet condition by recognizing the spike patterns of single units. Then we tested ANN responses for spike patterns recorded under various masker conditions. Addition of a masker generally altered spike patterns and disrupted ANN identification of signal location. That disruption was smaller, however, for signal and masker configurations in which the masker did not severely reduce units' spike rates. That result compared well with the psychophysical observation that listeners maintain good localization performance as long as signals are clearly audible. PMID- 11431505 TI - 5HT increases excitability of nociceptor-like rat dorsal root ganglion neurons via cAMP-coupled TTX-resistant Na(+) channels. AB - The physiological effects of 5HT receptor coupling to TTX-resistant Na(+) current, and the signaling pathway involved, was studied in a nociceptor-like subpopulation of rat dorsal root ganglion (DRG) cells (type 2), which can be identified by expression of a low-threshold, slowly inactivating A-current. The 5HT-mediated increase in TTX-resistant Na(+) current in type 2 DRG cells was mimicked and occluded by 10 microM forskolin. Superfusion of type 2 DRG cells on the outside with 1 mM 8-bromo-cAMP or chlorophenylthio-cAMP (CPT-cAMP) increased the Na(+) current, but less than 5HT itself. However, perfusion of the cells inside with 2 mM CPT-cAMP strongly increased the amplitude of control Na(+) currents and completely occluded the effect of 5HT. Thus it appears that the signaling pathway includes cAMP. The phosphodiesterase inhibitor 3-isobutyl-L methylxanthine (200 microM) also mimicked the effect of 5HT on Na(+) current, suggesting tonic adenylyl cyclase activity. 5HT reduced the amount of current required to evoke action potentials in type 2 DRG cells, suggesting that 5HT may lower the threshold for activation of nociceptor peripheral receptors. The above data suggest that serotonergic modulation of TTX-resistant Na(+) channels through a cAMP-dependent signaling pathway in nociceptors may participate in the generation of hyperalgesia. PMID- 11431506 TI - Effects of apomorphine on subthalamic nucleus and globus pallidus internus neurons in patients with Parkinson's disease. AB - This study examines the effect of apomorphine (APO), a nonselective D(1)- and D(2)-dopamine receptor agonist, on the firing activity of neurons in the subthalamic nucleus (STN) and internal segment of the globus pallidus (GPi) in patients with Parkinson's disease (PD). Single-unit microelectrode recordings were conducted in 13 patients undergoing implantation of deep brain stimulation electrodes in STN and 6 patients undergoing a pallidotomy. Doses of APO (2.5-8 mg) were sufficient to produce an ON state, but not intended to induce dyskinetic movements. Following baseline recordings from a single neuron, APO was administered and the activity of the neuron followed for an average of 15 min. The spontaneous discharge of neurons encountered before (n = 309), during (n = 146, 10-60 min), and after the effect of APO had waned (n = 127, >60 min) was also sampled, and the response to passive joint movements was noted. In both nuclei, APO increased the overall proportion of spikes in burst discharges (as detected with Poisson "surprise" analysis), and a greater proportion of cells with an irregular discharge pattern was observed. APO significantly decreased the overall firing rates of GPi neurons (P < 0.01), but there was no change in the overall firing rate of neurons in the STN (P = 0.68). However, the mean firing rates of STN neurons during APO-induced movements (choreic or dystonic dyskinesias) that occurred in four patients were significantly lower than OFF period baseline values (P < 0.05). Concurrent with a reduction in limb tremor, the percentage of cells with tremor-related activity (TCs) was found to be significantly reduced from 19 to 6% in the STN and 14 to 0% in the GPi following APO administration. APO also decreased the firing rate of STN TCs (P < 0.05). During the OFF state, more than 15% of neurons tested (STN = 93, GPi = 63) responded to passive movement of two or more joints. After APO, this proportion decreased significantly to 7% of STN cells and 4% of GPi cells (STN = 28, GPi = 26). These findings suggest that the APO-induced amelioration of parkinsonian symptoms is not solely due to a decrease in overall activity in the GPi or STN as predicted by the current model of basal ganglia function in PD. PMID- 11431507 TI - Contribution of the hyperpolarization-activated current (I(h)) to membrane potential and GABA release in hippocampal interneurons. AB - Intrinsic GABAergic interneurons provide inhibitory input to the principal neurons of the hippocampus. The majority of interneurons located in stratum oriens (s.o.) of the CA1 region express the hyperpolarization-activated cation current known as I(h). In an effort to elucidate the role of this current in regulating the baseline excitability of these neurons and its participation in the regulation of the release of GABA onto CA1 pyramidal neurons, we utilized whole cell electrophysiological recordings from both populations of cells. In voltage-clamp experiments, hyperpolarization of the interneuron membrane initiated a large inward current with an estimated activation threshold of 51.6 +/- 7.6 mV and a half-maximal voltage of -73.0 +/- 7.0 mV. This current was blocked by bath application of the I(h) inhibitors ZD 7288 (50 microM) or cesium (2 mM). Current-clamp experiments at the interneuron resting membrane potential ( 61.3 +/- 1.2 mV) revealed a significant hyperpolarization, a decrease in the rate of spontaneous action potential discharge, an increase in the cellular input resistance, and the elimination of rebound afterdepolarizations during blockade of I(h) with ZD 7288 (50 microM). The hyperpolarizing effect of ZD 7288 was also substantially larger in interneurons clamped near -80 mV using current injection through the pipette. In addition to neurons exhibiting I(h), recordings were obtained from a small population of s.o. interneurons that did not exhibit this current. These cells demonstrated resting membrane potentials that were significantly more negative (-73.6 +/- 5.5 mV) than those observed in neurons expressing I(h), suggesting that this current contributes to more depolarized membrane potentials in these cells. Recordings from postsynaptic pyramidal neurons demonstrated that blockade of I(h) with ZD 7288 caused a substantial reduction (approximately 43%) in the frequency of spontaneous action potential dependent inhibitory postsynaptic currents (IPSCs), without altering their average amplitude. However, miniature action-potential-independent IPSC frequency, amplitude, and decay kinetics were unaltered by ZD 7288. These data suggest that I(h) is active at the resting membrane potential in s.o. interneurons and as a result contributes to the spontaneous activity of these cells and to the tonic inhibition of CA1 pyramidal neurons in the hippocampus. PMID- 11431508 TI - Calcium channels in Xenopus spinal neurons differ in somas and presynaptic terminals. AB - Calcium channels play dual roles in cell signaling by promoting membrane depolarization and allowing entry of calcium ions. Patch-clamp recordings of calcium and calcium-dependent currents from the soma of Xenopus spinal neurons indicate key functional differences from those of presynaptic terminals. Both terminals and somas exhibit prominent high-voltage-activated (HVA) calcium current, but only the soma expresses additional low-voltage-activated (LVA) T type current. Further differences are reflected in the HVA current; N- and R-type channels are predominant in the soma while the terminal calcium current is composed principally of N type with smaller contribution by L- and R-type channels. Potential physiological significance for these different distributions of channel types may lie in the differential channel kinetics. Activation of somatic HVA calcium current occurs more slowly than HVA currents in terminals. Additionally, somatic LVA calcium current activates and deactivates much more slowly than any HVA calcium current. Fast-activating and -deactivating calcium current may be critical to processing the rapid exocytotic response in terminals, whereas slow LVA and HVA calcium currents may play a central role in shaping the somatic firing pattern. In support of different kinetic behavior between these two compartments, we find that somatic calcium current activates a prominent slow chloride current not observed in terminal recordings. This current activates in response to calcium entering through either LVA or HVA channels and likely functions as a modulator of excitability or synaptic input. The restriction of this channel type to the soma lends further support to the idea that differential expression of fast and slow channel types in these neurons is dictated by differences in signaling requirements for somatic and terminal compartments. PMID- 11431509 TI - Induction of long-term depression in cerebellar Purkinje cells requires a rapidly turned over protein. AB - Evidence is presented indicating that the induction of long-term depression (LTD) in Purkinje cells (PCs) requires a rapidly turned over protein(s) during a critical time period within 15 min after the onset of LTD-inducing stimulation and that synthesis of this protein is maintained by mRNAs supplied via transcription. LTD was induced in granule cell axon (GA)-to-PC synapses by stimulation of these synapses at 1 Hz for 5 min in conjunction with the climbing fibers (CFs) forming synapses on the same PCs and represented by a persistent reduction in the GA-induced excitatory postsynaptic potentials (EPSPs). Not only a prolonged but also a brief (5 min) pulse application of translational inhibitors (anisomycin, puromycin, or cycloheximide) effectively blocked the LTD induction. Pulses applied during the period from 30 min before to 10 min after the onset of conjunctive stimulation blocked the LTD induction, but those applied 15 min after were ineffective. The three translational inhibitors blocked the LTD induction similarly, suggesting that the effect is due to their common action of inhibiting protein synthesis. Infusion of a mRNA cap analogue (7-methyl GTP) into PCs also blocked LTD induction, ensuring that the postsynaptic protein synthesis within PCs is required for LTD induction. Transcriptional inhibitors, actinomycin D and 5,6-dichloro-l-beta-D-ribofuranosyl-benzimidazole, also blocked the LTD induction, but this effect was apparent when 5-min pulses of the transcriptional inhibitors preceded the conjunctive stimulation by 30 min or more. This time lag of 30 min is presumed to be required for depletion of the protein(s) required for LTD induction. The presently observed effects of translational and transcriptional inhibitors on the LTD induction are of temporal characteristics corresponding to their depressant effects on the type-1 metabotropic glutamate receptor (mGluR1)-mediated slow EPSPs in PCs as we have reported recently. An antagonist of mGluR1s [(RS)-1-aminoindan-1,5-dicarboxylic acid], however, did not block LTD induction when it was applied during the 10-min period following conjunctive stimulation, where translational inhibitors effectively blocked LTD induction. This discrepancy in time course suggests that the rapidly turned over protein(s) required for LTD induction is involved in a process occurring downstream of the activation of mGluR1s. PMID- 11431510 TI - Learning increases stimulus salience in anterior inferior temporal cortex of the macaque. AB - With experience, an object can become behaviorally relevant and thereby quickly attract our interest when presented in a visual scene. A likely site of these learning effects is anterior inferior temporal (aIT) cortex, where neurons are thought to participate in the filtering of irrelevant information out of complex visual displays. We trained monkeys to saccade consistently to one of two pictures in an array, in return for a reward. The array was constructed by pairing two stimuli, one of which elicited a good response from the cell when presented alone ("good" stimulus) and the other of which elicited a poor response ("poor" stimulus). The activity of aIT cells was recorded while monkeys learned to saccade to either the good or poor stimulus in the array. We found that neuronal responses to the array were greater (before the saccade occurred) when training reinforced a saccade to the good stimulus than when training reinforced a saccade to the poor stimulus. This difference was not present on incorrect trials, i.e., when saccades to the incorrect stimulus were made. Thus the difference in activity was correlated with performance. The response difference grew over the course of the recording session, in parallel with the improvement in performance. The response difference was not preceded by a difference in the baseline activity of the cells, unlike what was found in studies of cued visual search and working memory in aIT cortex. Furthermore, we found similar effects in a version of the task in which any of 10 possible pairs of stimuli, prelearned before the recording session, could appear on a given trial, thereby precluding a working memory strategy. The results suggest that increasing the behavioral significance of a stimulus through training alters the neural representation of that stimulus in aIT cortex. As a result, neurons responding to features of the relevant stimulus may suppress neurons responding to features of irrelevant stimuli. PMID- 11431511 TI - Nitric oxide modulates Ca(2+) channels in dorsal root ganglion neurons innervating rat urinary bladder. AB - The effect of a nitric oxide (NO) donor on high-voltage-activated Ca(2+) channel currents (I(Ca)) was examined using the whole cell patch-clamp technique in L(6) S(1) dorsal root ganglion (DRG) neurons innervating the urinary bladder. The neurons were labeled by axonal transport of a fluorescent dye, Fast Blue, injected into the bladder wall. Approximately 70% of bladder afferent neurons exhibited tetrodotoxin (TTX)-resistant action potentials (APs), and 93% of these neurons were sensitive to capsaicin, while the remaining neurons had TTX sensitive spikes and were insensitive to capsaicin. The peak current density of nimodipine-sensitive L-type Ca(2+) channels activated by depolarizing pulses (0 mV) from a holding potential of -60 mV was greater in bladder afferent neurons with TTX-resistant APs (39.2 pA/pF) than in bladder afferent neurons with TTX sensitive APs (28.9 pA/pF), while the current density of omega-conotoxin GVIA sensitive N-type Ca(2+) channels was similar (43-45 pA/pF) in both types of neurons. In both types of neurons, the NO donor, S-nitroso-N-acetylpenicillamine (SNAP) (500 microM), reversibly reduced (23.4-26.6%) the amplitude of I(Ca) elicited by depolarizing pulses to 0 mV from a holding potential of -60 mV. SNAP induced inhibition of I(Ca) was reduced by 90% in the presence of omega-conotoxin GVIA but was unaffected in the presence of nimodipine, indicating that NO-induced inhibition of I(Ca) is mainly confined to N-type Ca(2+) channels. Exposure of the neurons for 30 min to 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, 10 microM), an inhibitor of NO-stimulated guanylyl cyclase, prevented the SNAP induced reduction in I(Ca). Extracellular application of 8-bromo-cGMP (1 mM) mimicked the effects of NO donors by reducing the peak amplitude of I(Ca) (28.6% of reduction). Action potential configuration and firing frequency during depolarizing current pulses were not altered by the application of SNAP (500 microM) in bladder afferent neurons with TTX-resistant and -sensitive APs. These results indicate that NO acting via a cGMP signaling pathway can modulate N-type Ca(2+) channels in DRG neurons innervating the urinary bladder. PMID- 11431512 TI - Developmental changes in hyperpolarization-activated currents I(h) and I(K(IR)) in isolated rat intracardiac neurons. AB - The hyperpolarization-activated nonselective cation current, I(h), was investigated in neonatal and adult rat intracardiac neurons. I(h) was observed in all neurons studied and displayed slow time-dependent rectification. I(h) was isolated by blockade with external Cs(+) (2 mM) and was inhibited irreversibly by the bradycardic agent, ZD 7288. Current density of I(h) was approximately twofold greater in neurons from neonatal (-4.1 pA/pF at -130 mV) as compared with adult ( 2.3 pA/pF) rats; however, the reversal potential and activation parameters were unchanged. The reversal potential and amplitude of I(h) was sensitive to changes in external Na(+) and K(+) concentrations. An inwardly rectifying K(+) current, I(K(IR)), was also present in intracardiac neurons from adult but not neonatal rats and was blocked by extracellular Ba(2+). I(K(IR)) was present in approximately one-third of the adult intracardiac neurons studied, with a current density of -0.6 pA/pF at -130 mV. I(K(IR)) displayed rapid activation kinetics and no time-dependent rectification consistent with the rapidly activating, inward K(+) rectifier described in other mammalian autonomic neurons. I(K(IR)) was sensitive to changes in external K(+), whereby raising the external K(+) concentration from 3 to 15 mM shifted the reversal potential by approximately +36 mV. Substitution of external Na(+) had no effect on the reversal potential or amplitude of I(K(IR)). I(K(IR)) density increases as a function of postnatal development in a population of rat intracardiac neurons, which together with a concomitant decrease in I(h) may contribute to changes in the modulation of neuronal excitability in adult versus neonatal rat intracardiac ganglia. PMID- 11431513 TI - Chemical induction of mGluR5- and protein synthesis--dependent long-term depression in hippocampal area CA1. AB - Recent work has demonstrated that specific patterns of synaptic stimulation can induce long-term depression (LTD) in area CA1 that depends on activation of metabotropic glutamate receptors (mGluRs) and rapid protein synthesis. Here we show that the same form of synaptic modification can be induced by brief application of the selective mGluR agonist (RS)-3,5-dihydroxyphenylglycine (DHPG). DHPG-LTD 1) is a saturable form of synaptic plasticity, 2) requires mGluR5, 3) is mechanistically distinct from N-methyl-D-aspartate receptor (NMDAR) -dependent LTD, and 4) shares a common expression mechanism with protein synthesis-dependent LTD evoked using synaptic stimulation. DHPG-LTD should be useful for biochemical analysis of mGluR5- and protein synthesis-dependent synaptic modification. PMID- 11431514 TI - Sensory input directs spatial and temporal plasticity in primary auditory cortex. AB - The cortical representation of the sensory environment is continuously modified by experience. Changes in spatial (receptive field) and temporal response properties of cortical neurons underlie many forms of natural learning. The scale and direction of these changes appear to be determined by specific features of the behavioral tasks that evoke cortical plasticity. The neural mechanisms responsible for this differential plasticity remain unclear partly because important sensory and cognitive parameters differ among these tasks. In this report, we demonstrate that differential sensory experience directs differential plasticity using a single paradigm that eliminates the task-specific variables that have confounded direct comparison of previous studies. Electrical activation of the basal forebrain (BF) was used to gate cortical plasticity mechanisms. The auditory stimulus paired with BF stimulation was systematically varied to determine how several basic features of the sensory input direct plasticity in primary auditory cortex (A1) of adult rats. The distributed cortical response was reconstructed from a dense sampling of A1 neurons after 4 wk of BF-sound pairing. We have previously used this method to show that when a tone is paired with BF activation, the region of the cortical map responding to that tone frequency is specifically expanded. In this report, we demonstrate that receptive-field size is determined by features of the stimulus paired with BF activation. Specifically, receptive fields were narrowed or broadened as a systematic function of both carrier-frequency variability and the temporal modulation rate of paired acoustic stimuli. For example, the mean bandwidth of A1 neurons was increased (+60%) after pairing BF stimulation with a rapid train of tones and decreased (-25%) after pairing unmodulated tones of different frequencies. These effects are consistent with previous reports of receptive-field plasticity evoked by natural learning. The maximum cortical following rate and minimum response latency were also modified as a function of stimulus modulation rate and carrier frequency variability. The cortical response to a rapid train of tones was nearly doubled if BF stimulation was paired with rapid trains of random carrier frequency, while no following rate plasticity was observed if a single carrier frequency was used. Finally, we observed significant increases in response strength and total area of functionally defined A1 following BF activation paired with certain classes of stimuli and not others. These results indicate that the degree and direction of cortical plasticity of temporal and receptive-field selectivity are specified by the structure and schedule of inputs that co-occur with basal forebrain activation and suggest that the rules of cortical plasticity do not operate on each elemental stimulus feature independently of others. PMID- 11431515 TI - Temporal frequency of whisker movement. I. Representations in brain stem and thalamus. AB - How does processing of information change the internal representations used in subsequent stages of sensory pathways? To approach this question, we studied the representations of whisker movements in the lemniscal and paralemniscal pathways of the rat vibrissal system. We recently suggested that these two pathways encode movement frequency in different ways. We proposed that paralemniscal thalamocortical circuits, functioning as phase-locked loops (PLLs), translate temporally coded information into a rate code. Here we focus on the two major trigeminal nuclei of the brain stem, nucleus principalis and subnucleus interpolaris, and on their thalamic targets, the ventral posteromedial nucleus (VPM) and the medial division of the posterior nucleus (POm). This is the first study in which these brain stem and thalamic nuclei were explored together in the same animals and using the same stimuli. We studied both single- and multi-unit activity. We moved the whiskers both mechanically and by air puffs; here we present air-puff-induced movements because they are more similar to natural movements than movements induced by mechanical stimulations. We describe the basic properties of the responses in these brain stem and thalamic nuclei. The responses in both brain stem nuclei were similar; responses to air puffs were mostly tonic and followed the trajectory of whisker movement. The responses in the two thalamic nuclei were similar during low-frequency stimulations or during the first pulses of high-frequency stimulations, exhibiting more phasic responses than those of brain stem neurons. However, with frequencies >2 Hz, VPM and POm responses differed, generating different representations of the stimulus frequency. In the VPM, response amplitudes (instantaneous firing rates) and spike counts (total number of spikes per stimulus cycle) decreased as a function of the frequency. In the POm, latencies increased and spike count decreased as a function of the frequency. Having described the basic response properties in the four nuclei, we then focus on a specific test of our PLL hypothesis for coding in the paralemniscal pathway. We used short-duration air puffs, much shorter than whisker movements during natural whisking. The activity in this situation was consistent with the prediction we made on the basis of the PLL hypothesis. PMID- 11431516 TI - Temporal frequency of whisker movement. II. Laminar organization of cortical representations. AB - Part of the information obtained by rodent whiskers is carried by the frequency of their movement. In the thalamus of anesthetized rats, the whisker frequency is represented by two different coding schemes: by amplitude and spike count (i.e., response amplitudes and spike counts decrease as a function of frequency) in the lemniscal thalamus and by latency and spike count (latencies increase and spike counts decrease as a function of frequency) in the paralemniscal thalamus (see accompanying paper). Here we investigated neuronal representations of the whisker frequency in the primary somatosensory ("barrel") cortex of the anesthetized rat, which receives its input from both the lemniscal and paralemniscal thalamic nuclei. Single and multi-units were recorded from layers 2/3, 4 (barrels only), 5a, and 5b during vibrissal stimulation. Typically, the input frequency was represented by amplitude and spike count in the barrels of layer 4 and in layer 5b (the "lemniscal layers") and by latency and spike count in layer 5a (the "paralemniscal layer"). Neurons of layer 2/3 displayed a mixture of the two coding schemes. When the pulse width of the stimulus was reduced from 50 to 20 ms, the latency coding in layers 5a and 2/3 was dramatically reduced, while the spike-count coding was not affected; in contrast, in layers 4 and 5b, the latencies remained constant, but the spike counts were reduced with 20-ms stimuli. The same effects were found in the paralemniscal and lemniscal thalamic nuclei, respectively (see accompanying paper). These results are consistent with the idea that thalamocortical loops of different pathways, although terminating within the same cortical columns, perform different computations in parallel. Furthermore, the mixture of coding schemes in layer 2/3 might reflect an integration of lemniscal and paralemniscal outputs. PMID- 11431517 TI - Distinct patterns of brain oscillations underlie two basic parameters of human maze learning. AB - We examine how oscillations in the intracranial electroencephalogram (iEEG) relate to human maze learning. Theta- band activity (4-12 Hz in rodents; 4-8 Hz in humans) plays a significant role in memory function in rodents and in humans. Recording intracranially in humans, we have reported task-related, theta-band rhythmic activity in the raw trace during virtual maze learning and during a nonspatial working memory task. Here we analyze oscillations during virtual maze learning across a much broader range of frequencies and analyze their relationship to two task variables relevant to learning. We describe a new algorithm for detecting oscillatory episodes that takes advantage of the high signal-to-noise ratio and high temporal resolution of the iEEG. Accounting for the background power spectrum of the iEEG, the algorithm allows us to directly compare levels of oscillatory activity across frequencies within the 2- to 45-Hz band. We report that while episodes of oscillatory activity are found at various frequencies, most of the rhythmic activity during virtual maze learning occurs within the theta band. Theta oscillations are more prevalent when the task is made more difficult (manipulation of maze length). However, these oscillations do not tend to covary significantly with decision time, a good index of encoding and retrieval operations. In contrast, lower- and higher-frequency oscillations do covary with this variable. These results suggest that while human cortically recorded theta might play a role in encoding, the overall levels of theta oscillations tell us little about the immediate demands on encoding or retrieval. Finally, different patterns of oscillations may reflect distinct underlying aspects of memory function. PMID- 11431518 TI - Imaging of 4-AP-induced, GABA(A)-dependent spontaneous synchronized activity mediated by the hippocampal interneuron network. AB - Under conditions of increased excitability, such as application of the K(+) channel blocker 4-aminopyridine (4-AP, 100 microM), interneurons in the hippocampal slice show an additional form of synchronized activity that is distinct from the ictal and interictal epileptiform activity induced by these manipulations. In principal neurons, i.e., pyramidal and granule cells, this synchronized interneuron activity (SIA) generates large, multi-component synaptic potentials, which have been termed long-lasting depolarizations (LLDs). These LLDs are dependent on GABA(A) receptor-mediated synaptic transmission but not on excitatory amino acid (EAA) receptors. Intracellular recordings from hilar interneurons have shown that depolarizing GABA(A) receptor-mediated synaptic potentials are also largely responsible for the synchronization of interneurons. The spatiotemporal characteristics of this interneuron activity have not been investigated previously. Using a voltage-sensitive dye and optical techniques that are capable of recording spontaneous synchronized activity, we have characterized the spatiotemporal pattern of SIA (in the presence of 4-AP + EAA receptor antagonists) and compared it with interictal epileptiform activity (in 4 AP only). Like interictal activity, SIA could be observed throughout the hippocampal slice. Unlike interictal activity, which originated in area CA2/CA3 and spread from there, SIA was most prominent in area CA1 and originated either there or in the subiculum. In CA1, interictal activity was largest in and near stratum pyramidale, while SIA was mainly located in s. lacunosum moleculare. Furthermore SIA was equally likely to propagate in either direction, and multiple patterns of propagation could be observed within a single brain slice. These studies suggest that hippocampal area CA1 has the highest propensity for SIA, that multiple locations can serve as the site of origin, and that interneurons located in s. lacunosum moleculare or interneurons that specifically project to this region may be particularly important for synchronized interneuron activity. PMID- 11431519 TI - Developmental changes in trial-to-trial variations in whisker barrel responses studied using intrinsic optical imaging: comparison between normal and de whiskered rats. AB - We used an intrinsic optical imaging technique to examine postnatal developmental changes in the rat barrel response to a single whisker movement. We compared the optical response patterns between control and de-whiskered rats, from which whiskers were removed except for the D1 whisker just after birth. Barrel responses were evoked by D1-whisker movement stimulation, and the intrinsic optical signals were detected from the somatosensory cortex through the dura mater. In the control rats, the area of the barrel response decreased gradually as postnatal development proceeded from 2 to 7 wk, until reaching the adult pattern. On the other hand, in the de-whiskered rats, the barrel response area did not change during development and showed a larger size than in the control rats. We also compared the trial-to-trial variations in the barrel responses between the two groups. In the control rats, trial-to-trial variations in the optical responses were observed under the same conditions of whisker stimulation, and the extent of the variations decreased with postnatal development up to 7 wk. In the de-whiskered rats, trial-to-trial variations were also observed, but the extent was larger and unchanged during development. In both groups, the positions of the response area were the same with respect to the bregma. These results suggest that the decrease in the area and variations in the optical responses are caused by interactions of the corresponding whisker barrel with neighboring barrels and that these interactions are necessary for the developmental stabilization of the intracortical horizontal connections, which are widespread and have high plasticity in early postnatal periods. PMID- 11431520 TI - Cortical representation of the sensory dimension of pain. AB - It is well accepted that pain is a multidimensional experience, but little is known of how the brain represents these dimensions. We used positron emission tomography (PET) to indirectly measure pain-evoked cerebral activity before and after hypnotic suggestions were given to modulate the perceived intensity of a painful stimulus. These techniques were similar to those of a previous study in which we gave suggestions to modulate the perceived unpleasantness of a noxious stimulus. Ten volunteers were scanned while tonic warm and noxious heat stimuli were presented to the hand during four experimental conditions: alert control, hypnosis control, hypnotic suggestions for increased-pain intensity and hypnotic suggestions for decreased-pain intensity. As shown in previous brain imaging studies, noxious thermal stimuli presented during the alert and hypnosis-control conditions reliably activated contralateral structures, including primary somatosensory cortex (S1), secondary somatosensory cortex (S2), anterior cingulate cortex, and insular cortex. Hypnotic modulation of the intensity of the pain sensation led to significant changes in pain-evoked activity within S1 in contrast to our previous study in which specific modulation of pain unpleasantness (affect), independent of pain intensity, produced specific changes within the ACC. This double dissociation of cortical modulation indicates a relative specialization of the sensory and the classical limbic cortical areas in the processing of the sensory and affective dimensions of pain. PMID- 11431521 TI - Intrinsic optical signals in respiratory brain stem regions of mice: neurotransmitters, neuromodulators, and metabolic stress. AB - In the rhythmic brain stem slice preparation, spontaneous respiratory activity is generated endogenously and can be recorded as output activity from hypoglossal XII rootlets. Here we combine these recordings with measurements of the intrinsic optical signal (IOS) of cells in the regions of the periambigual region and nucleus hypoglossus of the rhythmic slice preparation. The IOS, which reflects changes of infrared light transmittance and scattering, has been previously employed as an indirect sensor for activity-related changes in cell metabolism. The IOS is believed to be primarily caused by cell volume changes, but it has also been associated with other morphological changes such as dendritic beading during prolonged neuronal excitation or mitochondrial swelling. An increase of the extracellular K(+) concentration from 3 to 9 mM, as well as superfusion with hypotonic solution induced a marked increase of the IOS, whereas a decrease in extracellular K(+) or superfusion with hypertonic solution had the opposite effect. During tissue anoxia, elicited by superfusion of N(2)-gassed solution, the biphasic response of the respiratory activity was accompanied by a continuous rise in the IOS. On reoxygenation, the IOS returned to control levels. Cells located at the surface of the slice were observed to swell during periods of anoxia. The region of the nucleus hypoglossus exhibited faster and larger IOS changes than the periambigual region, which presumably reflects differences in sensitivities of these neurons to metabolic stress. To analyze the components of the hypoxic IOS response, we investigated the IOS after application of neurotransmitters known to be released in increasing amounts during hypoxia. Indeed, glutamate application induced an IOS increase, whereas adenosine slightly reduced the IOS. The IOS response to hypoxia was diminished after application of glutamate uptake blockers, indicating that glutamate contributes to the hypoxic IOS. Blockade of the Na(+)/K(+)-ATPase by ouabain did not provoke a hypoxia-like IOS change. The influences of K(ATP) channels were analyzed, because they contribute significantly to the modulation of neuronal excitability during hypoxia. IOS responses obtained during manipulation of K(ATP) channel activity could be explained only by implicating mitochondrial volume changes mediated by mitochondrial K(ATP) channels. In conclusion, the hypoxic IOS response can be interpreted as a result of cell and mitochondrial swelling. Cell swelling can be attributed to hypoxic release of neurotransmitters and neuromodulators and to inhibition of Na(+)/K(+)-pump activity. PMID- 11431522 TI - Acetylcholine-dependent induction and expression of functional plasticity in the barrel cortex of the adult rat. AB - The involvement of acetylcholine (ACh) in the induction of neuronal sensory plasticity is well documented. Recently we demonstrated in the somatosensory cortex of the anesthetized rat that ACh is also involved in the expression of neuronal plasticity. Pairing stimulation of the principal whisker at a fixed temporal frequency with ACh iontophoresis induced potentiations of response that required re-application of ACh to be expressed. Here we fully characterize this phenomenon and extend it to stimulation of adjacent whiskers. We show that these ACh-dependent potentiations are cumulative and reversible. When several sensori cholinergic pairings were applied consecutively with stimulation of the principal whisker, the response at the paired frequency was further increased, demonstrating a cumulative process that could reach saturation levels. The potentiations were specific to the stimulus frequency: if the successive pairings were done at different frequencies, then the potentiation caused by the first pairing was depotentiated, whereas the response to the newly paired frequency was potentiated. During testing, the potentiation of response did not develop immediately on the presentation of the paired frequency during application of ACh: the analysis of the kinetics of the effect indicates that this process requires the sequential presentation of several trains of stimulation at the paired frequency to be expressed. We present evidence that a plasticity with similar characteristics can be induced for responses to stimulation of an adjacent whisker, suggesting that this potentiation could participate in receptive field spatial reorganizations. The spatial and temporal properties of the ACh-dependent plasticity presented here impose specific constraints on the underlying cellular and molecular mechanisms. PMID- 11431523 TI - Pre-Botzinger neurons with preinspiratory discharges "in vivo" express NK1 receptors in the rat. AB - Substance P stimulates respiration, in part by a direct action on the pre Botzinger complex (preBotC). This region of the medulla oblongata contains neurons that are strongly immunoreactive for the neurokinin-1 receptor (NK1R-ir), and a recent theory has postulated that these cells might be the adult form of excitatory interneurons that are essential for respiratory rhythmogenesis in neonates. Here we sought to determine whether preBotC respiratory neurons are indeed NK1R-ir in the adult rat. Preinspiratory (pre-I) neurons were recorded in the preBotC region of halothane-anesthetized rats. Most pre-I cells could be antidromically activated from the contralateral side of the medulla (7 of 10; latency 1.3 +/- 0.2 ms), suggesting that most of them were propriomedullary neurons rather than respiratory motoneurons or bulbospinal cells. Thirty-two pre I neurons including seven cells with contralateral projection were labeled with biotinamide using the juxtacellular method. Eleven cells (34.4%) were NK1R-ir, including three of the seven pre-I cells that were antidromically activated from the contralateral side. In 3 control rats we labeled 20 preBotC neurons with patterns of discharge other than pre-I and found that none were detectably NK1R ir. In conclusion, some of the intensely NK1R-ir neurons of the adult preBotC region are indeed respiratory interneurons as suggested by Gray et al. The subtype of NK1R identified by the antibody is detectable only in a small minority of preBotC respiratory cells, most notably in pre-I interneurons. Given prior anatomical evidence, these NK1R-ir pre-I interneurons are most likely glutamatergic. The data are consistent with the possibility that the NK1R-ir pre I interneurons of the adult preBotC could be the adult form of a class of inspiratory neurons that are rhythmogenic in the neonate (either the pacemakers and/or an excitatory subtype of follower neurons). PMID- 11431524 TI - Patterns of locomotor drive to motoneurons and last-order interneurons: clues to the structure of the CPG. AB - We have examined the linkage between patterns of activity in several hindlimb motor pools and the modulation of oligosynaptic cutaneous reflex pathways during fictive locomotion in decerebrate unanesthetized cats to assess the notion that such linkages can shed light on the structure of the central pattern generator (CPG) for locomotion. We have concentrated attention on the cutaneous reflex pathways that project to the flexor digitorum longus (FDL) motor pool because of that muscle's unique variable behavior during normal and fictive locomotion in the cat. Differential locomotor control of last-order excitatory interneurons in pathways from low-threshold cutaneous afferents in the superficial peroneal and medial plantar afferents to FDL motoneurons is fully documented for the first time. The qualitative patterns of differential control are shown to remain the same whether the FDL muscle is active in early flexion, as usually found, or during the extension phase of fictive locomotion, which is less common during fictive stepping. The patterns of motor pool activity and of reflex pathway modulation indicate that the flexion phase of fictive locomotion has distinct early versus late components. Observations during "normal" and unusual patterns of fictive stepping suggest that some aspects of locomotor pattern formation can be separated from rhythm generation, implying that these two CPG functions may be embodied, at least in part, in distinct neural organizations. The results are discussed in relation to a provisional circuit diagram that could explain the experimental findings. PMID- 11431525 TI - Nicotine-induced enhancement of glutamatergic and GABAergic synaptic transmission in the mouse amygdala. AB - Presynaptic nicotinic acetylcholine receptors (nAChRs) are thought to mediate some of the cognitive and behavioral effects of nicotine. The olfactory projection to the amygdala, and intra-amygdaloid projections, are limbic relays involved in behavioral reinforcement, a property influenced by nicotine. Co cultures consisting of murine olfactory bulb (OB) explants and dispersed amygdala neurons were developed to reconstruct this pathway in vitro. Whole cell patch clamp recordings were obtained from amygdala neurons contacted by OB explant neurites, and spontaneous and evoked synaptic currents were characterized. The majority of the 108 innervated amygdala neurons exhibited glutamatergic spontaneous postsynaptic currents (PSCs), 20% exhibited GABAergic spontaneous PSCs, and 17% exhibited both. Direct extracellular stimulation of OB explants elicited glutamatergic synaptic currents in amygdala neurons. Antibodies to nAChR subunits co-localized with an antibody to synapsin I, a presynaptic marker, along OB explant processes, consistent with the targeting of nAChR protein to presynaptic sites of the mitral cell projections. Hence, we examined the role of presynaptic nAChRs in modulating synaptic transmission in the OB-amygdala co cultures. Focal application of 500 nM to 1 microM nicotine for 5-60 s markedly increased the frequency of spontaneous PSCs, without a change in the amplitude, in 39% of neurons that exhibited glutamatergic spontaneous PSCs (average peak fold increase = 125.2 +/- 33.3). Nicotine also enhanced evoked glutamatergic currents elicited by direct stimulation of OB explant fibers. Nicotine increased the frequency of spontaneous PSCs, without a change in the amplitude, in 35% of neurons that exhibited GABAergic spontaneous PSCs (average peak fold increase = 63.9 +/- 34.3). Thus activation of presynaptic nAChRs can modulate glutamatergic as well as GABAergic synaptic transmission in the amygdala. These results suggest that behaviors mediated by olfactory projections may be modulated by presynaptic nAChRs in the amygdala, where integration of olfactory and pheromonal input is thought to occur. PMID- 11431526 TI - Spectrotemporal organization of excitatory and inhibitory receptive fields of cat posterior auditory field neurons. AB - The excitatory and inhibitory frequency/intensity response areas (FRAs) and spectrotemporal receptive fields (STRFs) of posterior auditory cortical field (PAF) single neurons were investigated in barbiturate anesthetized cats. PAF neurons' pure-tone excitatory FRAs (eFRAs) exhibited a diversity of shapes, including some with very broad frequency tuning and some with multiple distinct excitatory frequency ranges (i.e., multipeaked eFRAs). Excitatory FRAs were analyzed after selectively excluding spikes on the basis of spike response times relative to stimulus onset. This analysis indicated that spikes with shorter response times were confined to narrow regions of the eFRAs, while spikes with longer response times were more broadly distributed over the eFRA. First-spike latencies in higher threshold response peaks of multipeaked eFRAs were approximately 10 ms longer, on average, than latencies in lower threshold response peaks. STRFs were constructed to examine the dynamic frequency tuning of neurons. More than half of the neurons (51%) had STRFs with "sloped" response maxima, indicating that the excitatory frequency range shifted with time. A population analysis demonstrated that the median first-spike latency varied systematically as a function of frequency with a median slope of approximately 12 ms per octave. Inhibitory frequency response areas were determined by simultaneous two-tone stimulation. As in primary auditory cortex (A1), a diversity of inhibitory band structures was observed. The largest class of neurons (25%) had an inhibitory band flanking each eFRA edge, i.e., one lower and one upper inhibitory band in a "center-surround" organization. However, in comparison to a previous report of inhibitory structure in A1 neurons, PAF exhibited a higher incidence of neurons with more complex inhibitory band structure (for example, >2 inhibitory bands). As was the case with eFRAs, spikes with longer response times contributed to the complexity of inhibitory FRAs. These data indicate that PAF neurons integrate temporally varying excitatory and inhibitory inputs from a broad spectral extent and, compared with A1, may be suited to analyzing acoustic signals of greater spectrotemporal complexity than was previously thought. PMID- 11431527 TI - Transition from GABAergic to glycinergic synaptic transmission in newly formed spinal networks. AB - The role of glycinergic and GABAergic systems in mediating spontaneous synaptic transmission in newly formed neural networks was examined in motoneurons in the developing rat spinal cord. Properties of action potential-independent miniature inhibitory postsynaptic currents (mIPSCs) mediated by glycine and GABA(A) receptors (GlyR and GABA(A)R) were studied in spinal cord slices of 17- to 18-day old embryos (E17-18) and 1- to 3-day-old postnatal rats (P1-3). mIPSC frequency and amplitude significantly increased after birth, while their decay time decreased. To determine the contribution of glycinergic and GABAergic synapses to those changes, GlyR- and GABA(A)R-mediated mIPSCs were isolated based on their pharmacological properties. Two populations of pharmacologically distinct mIPSCs were recorded in the presence of glycine or GABA(A) receptors antagonists: bicuculline-resistant, fast-decaying GlyR-mediated mIPSCs, and strychnine resistant, slow-decaying GABA(A)R-mediated mIPSCs. The frequency of GABA(A)R mediated mIPSCs was fourfold higher than that of GlyR-mediated mIPSCs at E17-18, indicating that GABAergic synaptic sites were functionally dominant at early stages of neural network formation. Properties of GABA(A)R-mediated mIPSC amplitude fluctuations changed from primarily unimodal skewed distribution at E17 18 to Gaussian mixtures with two to three discrete components at P1-3. A developmental shift from primarily long-duration GABAergic mIPSCs to short duration glycinergic mIPSCs was evident after birth, when the frequency of GlyR mediated mIPSCs increased 10-fold. This finding suggested that either the number of glycinergic synapses or the probability of vesicular glycine release increased during the period studied. The increased frequency of GlyR-mediated mIPSCs was associated with more than a twofold increase in their mean amplitude, and in the number of motoneurons in which mIPSC amplitude fluctuations were best fitted by multi-component Gaussian curves. A third subpopulation of mIPSCs was apparent in the absence of glycine and GABA(A) receptor antagonists: mIPSCs with both fast and slow decaying components. Based on their dual-component decay time and their suppression by either strychnine or bicuculline, we assumed that these were generated by the activation of co-localized postsynaptic glycine and GABA(A) receptors. The contribution of mixed glycine-GABA synaptic sites to the generation of mIPSCs did not change after birth. The developmental switch from predominantly long-duration GABAergic inhibitory synaptic currents to short duration glycinergic currents might serve as a mechanism regulating neuronal excitation in the developing spinal networks. PMID- 11431528 TI - Initiation and propagation of regenerative Ca(2+)-dependent potentials in dendrites of layer 5 pyramidal neurons. AB - The initiation and propagation of dendritic Ca(2+)-dependent regenerative potentials (CDRPs) were investigated by imaging the Ca(2+)-sensitive dye Fluo-4 during whole cell recording from the soma of layer 5 pyramidal neurons visualized in a slice preparation of rat neocortex by the use of infrared-differential interference contrast microscopy. CDRPs were evoked by focal iontophoresis of glutamate at visually identified sites 178-648 microm from the soma on the apical dendrite and at sites on the basal dendrites. Increases in [Ca(2+)](i) were maximal near the site of iontophoresis and were graded with iontophoretic current that was subthreshold for evoking CDRPs. CDRP initiation was associated with a [Ca(2+)](i) rise that differed from a just-subthreshold response in both magnitude and spatial extent but whose amplitude declined both proximal and distal to the iontophoretic site. These [Ca(2+)](i) rises, whether associated with subthreshold or regenerative voltage responses, were minimally affected by blockade of N-methyl-D-aspartate receptors but were abolished by Cd(2+), suggesting that Ca(2+) influx through voltage-gated channels caused the rise of [Ca(2+)](i). On the assumption that the rise of [Ca(2+)](i) during a CDRP marks the spatial extent of regenerative Ca(2+) influx, we conclude that CDRPs can be evoked at any point on the main apical or basal trunk where membrane potential reaches CDRP threshold rather than at discrete "hot spots," the CDRP is initiated at a spatially restricted site, and it propagates decrementally both distal and proximal to its initiation site. These results raise the possibility that synaptic integration may occur first in the dendrites to evoke a CDRP. Because these responses propagate decrementally to the soma, they are able to sum with input from other regions of the cell so that the cell as a whole remains integrative. PMID- 11431529 TI - Dendritic calcium spikes in layer 5 pyramidal neurons amplify and limit transmission of ligand-gated dendritic current to soma. AB - Long-lasting, dendritic, Ca(2+)-dependent action potentials (plateaus) were investigated in layer 5 pyramidal neurons from rat neocortical slices visualized by infrared-differential interference contrast microscopy to understand the role of dendritic Ca(2+) spikes in the integration of synaptic input. Focal glutamate iontophoresis on visualized dendrites caused soma firing rate to increase linearly with iontophoretic current until dendritic Ca(2+) responses caused a jump in firing rate. Increases in iontophoretic current caused no further increase in somatic firing rate. This limitation of firing rate resulted from the inability of increased glutamate to change evoked plateau amplitude. Similar nonlinear patterns of soma firing were evoked by focal iontophoresis on the distal apical, oblique, and basal dendrites, whereas iontophoresis on the soma and proximal apical dendrite only evoked a linear increase in firing rate as a function of iontophoretic current without plateaus. Plateau amplitude recorded in the soma decreased as the site of iontophoresis was moved farther from the soma, consistent with decremental propagation of the plateau to the soma. Currents arriving at the soma summed if plateaus were evoked on separate dendrites or if subthreshold responses were evoked from sites on the same dendrite. If plateaus were evoked at two sites on the same dendrite, only the proximal plateau was seen at the soma. Just-subthreshold depolarizations at two sites on the same dendrite could sum to evoke a plateau at the proximal site. We conclude that the plateaus prevent current from ligand-gated channels distal to the plateau-generating region from reaching the soma and directly influencing firing rate. The implications of plateau properties for synaptic integration are discussed. PMID- 11431530 TI - Dendritic mechanisms of phase precession in hippocampal CA1 pyramidal neurons. AB - Dual whole-cell patch clamp recordings from the soma and dendrites of CA1 pyramidal neurons located in hippocampal slices of adult rats were used to examine the potential mechanisms of phase precession. To mimic phasic synaptic input, 5-Hz sine wave current injections were simultaneously delivered both to the soma and apical dendrites (dendritic current was 180 degrees out-of-phase with soma). Increasing the amplitude of the dendritic current injection caused somatic action potential initiation to advance in time (move forward up to 180 degrees). The exact pattern of phase advancement is dependent on the dendritic location of input, with more distal input causing a more gradual temporal shift in spike initiation and a smaller increase in spike number. Patterned stimulation of Schaffer collateral/perforant path synaptic input can produce phase precession that is very similar to that observed with sine wave current injections. Finally, the exact amount of synaptic input required to produce phase advancement was found to be regulated by dendritic voltage-gated ion channels. Together, these data demonstrate that the summation of primarily proximal inhibition with an increasing amount of out-of-phase, primarily distal excitation can result in a form of phase advancement similar to that seen during theta activity in the intact hippocampus. PMID- 11431531 TI - Comparison of the effects of stimulating groups of static gamma axons with different conduction velocity ranges on cat spindles. AB - In cat peroneus tertius muscles, static gamma axons were prepared in groups of three to four according to the conduction velocity of their axons (fast, intermediate, or slow). Effects of stimulating these groups (at 20, 30, and 50 Hz) on spindle ensemble discharges during sinusoidal stretch (peak-to-peak amplitude, 0.5 mm; frequency linearly increasing from 0.5 to 8 Hz in 10 s) were compared. Ensemble discharges were obtained by digital treatment of the discharges in afferent fibers from all the spindles in peroneus tertius as recorded from the muscle nerve. Stimulation of each group prevented ensemble discharges from falling to very low levels during shortening phases. However, this effect was clearly larger when the group of fast-conducting axons was stimulated. In view of the known effects of the activation of bag(2) and chain fibers (either separately or together) on single primary ending discharges during comparable sinusoidal stretches, this stronger effect supports the view that static gamma axons with faster conduction velocities are more likely to supply more bag(2) fibers than slower ones. Possibly the proportions of bag(2) and chain fibers activated during motor activity are determined by a recruitment of static gamma motoneurons related to their size. PMID- 11431532 TI - GABA(B) and Trk receptor signaling mediates long-lasting inhibitory synaptic depression. AB - In many areas of the nervous system, excitatory and inhibitory synapses are reconfigured during early development. We have previously described the anatomical refinement of an inhibitory projection from the medial nucleus of the trapezoid body to the lateral superior olive in the developing gerbil auditory brain stem. Furthermore, these inhibitory synapses display an age-dependent form of long-lasting depression when activated at a low rate, suggesting that this process could support inhibitory synaptic refinement. Since the inhibitory synapses release both glycine and GABA during maturation, we tested whether GABA(B) receptor signaling could initiate the decrease in synaptic strength. When whole cell recordings were made from lateral superior olive neurons in a brain slice preparation, the long-lasting depression of medial nucleus of the trapezoid body-evoked inhibitory potentials was eliminated by the GABA(B) receptor antagonist, SCH-50911. In addition, inhibitory potentials could be depressed by repeated exposure to the GABA(B) receptor agonist, baclofen. Since GABA(B) receptor signaling may not account entirely for inhibitory synaptic depression, we examined the influence of neurotrophin signaling pathways located in the developing superior olive. Bath application of brain-derived neurotrophic factor or neurotrophin-3 depressed evoked inhibitory potentials, and use-dependent depression was blocked by the tyrosine kinase antagonist, K-252a. We suggest that early expression of GABAergic and neurotrophin signaling mediates inhibitory synaptic plasticity, and this mechanism may support the anatomical refinement of inhibitory connections. PMID- 11431533 TI - Ubiquitination of a new form of alpha-synuclein by parkin from human brain: implications for Parkinson's disease. AB - Parkinson's disease (PD) is a common neurodegenerative disorder characterized by the progressive accumulation in selected neurons of protein inclusions containing alpha-synuclein and ubiquitin. Rare inherited forms of PD are caused by autosomal dominant mutations in alpha-synuclein or by autosomal recessive mutations in parkin, an E3 ubiquitin ligase. We hypothesized that these two gene products interact functionally, namely, that parkin ubiquitinates alpha-synuclein normally and that this process is altered in autosomal recessive PD. We have now identified a protein complex in normal human brain that includes parkin as the E3 ubiquitin ligase, UbcH7 as its associated E2 ubiquitin conjugating enzyme, and a new 22-kilodalton glycosylated form of alpha-synuclein (alphaSp22) as its substrate. In contrast to normal parkin, mutant parkin associated with autosomal recessive PD failed to bind alphaSp22. In an in vitro ubiquitination assay, alphaSp22 was modified by normal but not mutant parkin into polyubiquitinated, high molecular weight species. Accordingly, alphaSp22 accumulated in a non ubiquitinated form in parkin-deficient PD brains. We conclude that alphaSp22 is a substrate for parkin's ubiquitin ligase activity in normal human brain and that loss of parkin function causes pathological alphaSp22 accumulation. These findings demonstrate a critical biochemical reaction between the two PD-linked gene products and suggest that this reaction underlies the accumulation of ubiquitinated alpha-synuclein in conventional PD. PMID- 11431534 TI - Observation of quantum shock waves created with ultra- compressed slow light pulses in a Bose-Einstein condensate. AB - We have used an extension of our slow light technique to provide a method for inducing small density defects in a Bose-Einstein condensate. These sub- resolution, micrometer-sized defects evolve into large-amplitude sound waves. We present an experimental observation and theoretical investigation of the resulting breakdown of superfluidity, and we observe directly the decay of the narrow density defects into solitons, the onset of the "snake" instability, and the subsequent nucleation of vortices. PMID- 11431535 TI - Mid-Holocene climate in the south-central Andes: humid or dry? PMID- 11431536 TI - New world pathogen strategy disclosed. PMID- 11431537 TI - White House. President's new adviser ready to put science in its place. PMID- 11431538 TI - Science policy. Bush grapples with stem cells, cloning. PMID- 11431539 TI - Science publishing. Lab chief, postdoc clash over nanotech paper. PMID- 11431540 TI - Ultrafast lasers. Lighting the way to a quantum computer. PMID- 11431541 TI - Cell biology. NO helps make fireflies flash. PMID- 11431542 TI - Astrophysics. 'Tired-light' hypothesis gets re-tired. PMID- 11431543 TI - Particle physics. Russian turmoil rattles CERN. PMID- 11431544 TI - Cell cycle research. DNA: once copied, thrice blocked. PMID- 11431545 TI - Archaeology. Unwrapping a modern mummy mystery. PMID- 11431546 TI - Archaeology. Writing gets a rewrite. PMID- 11431547 TI - Profile. Dollars and cents vs. the AIDS epidemic. PMID- 11431548 TI - Ecology. Dammed if you do, damned if you don't? PMID- 11431549 TI - European research area. E.U. research chief's strategy: unite and conquer. PMID- 11431550 TI - American Astronomical Society meeting. Dust disks and black holes swell the cosmic census. PMID- 11431551 TI - Ecology. A new wind sweeps the plains. PMID- 11431552 TI - Signal transduction: signaling specificity- a complex affair. PMID- 11431553 TI - Cosmology. Magnetic mysteries. PMID- 11431554 TI - Evolution. Haploids--hapless or happening? PMID- 11431555 TI - Paleoclimate. Extending the calibrated radiocarbon record. PMID- 11431556 TI - Ecology. Marine bugs and carbon flow. PMID- 11431557 TI - Plate tectonics. A graveyard for buoyant slabs? PMID- 11431558 TI - Graded materials for resistance to contact deformation and damage. AB - The mechanical response of materials with spatial gradients in composition and structure is of considerable interest in disciplines as diverse as tribology, geology, optoelectronics, biomechanics, fracture mechanics, and nanotechnology. The damage and failure resistance of surfaces to normal and sliding contact or impact can be changed substantially through such gradients. This review assesses the current understanding of the resistance of graded materials to contact deformation and damage, and outlines future research directions and possible applications for graded materials. PMID- 11431559 TI - Ultrafast manipulation of electron spin coherence. AB - A technique is developed with the potential for coherent all-optical control over electron spins in semiconductors on femtosecond time scales. The experiments show that optical "tipping" pulses can enact substantial rotations of electron spins through a mechanism dependent on the optical Stark effect. These rotations were measured as changes in the amplitude of spin precession after optical excitation in a transverse magnetic field and approach pi/2 radians. A prototype sequence of two tipping pulses indicates that the rotation is reversible, a result that establishes the coherent nature of the tipping process. PMID- 11431560 TI - Superconductivity in 4 angstrom single-walled carbon nanotubes. AB - Investigation of the magnetic and transport properties of single-walled small diameter carbon nanotubes embedded in a zeolite matrix revealed that at temperatures below 20 kelvin, 4 angstrom tubes exhibit superconducting behavior manifest as an anisotropic Meissner effect, with a superconducting gap and fluctuation supercurrent. The measured superconducting characteristics display smooth temperature variations owing to one-dimensional fluctuations, with a mean field superconducting transition temperature of 15 kelvin. Statistical mechanic calculations based on the Ginzburg-Landau free-energy functional yield predictions that are in excellent agreement with the experiments. PMID- 11431561 TI - Construction principles of "hyparenes": families of molecules with planar pentacoordinate carbons. AB - Density-functional theory calculations predict that three borocarbon units with planar pentacoordinate carbons -C3B3-, -C2B4-, and -CB5-, can replace the -(CH)3- subunits in aromatic or even in antiaromatic hydrocarbons to construct "hyparenes" (families of molecules with planar pentacoordinate carbons). These borocarbon units contribute two, one, and zero electrons, respectively, to the parent pi system. Depending on the choice of these units, the hyparenes (judging from computed proton and nucleus-independent chemical shifts), can maintain or can interconvert the aromatic or antiaromatic character of the parent compounds. The hyparenes are low-lying local minima with normal carbon-boron, boron-boron, and carbon-carbon bond lengths. The multicenter bonding in the hyparenes involves contributions of partial sigma and partial pi bonds to the planar pentacoordinate carbons; the octet rule is not violated. Borocarbon species, for which there is some mass spectrometric evidence, might be observed and identified, for example, in matrix isolation by vibrational spectroscopy. PMID- 11431562 TI - Predicting the mesophases of copolymer-nanoparticle composites. AB - The interactions between mesophase-forming copolymers and nanoscopic particles can lead to highly organized hybrid materials. The morphology of such composites depends not only on the characteristics of the copolymers, but also on the features of the nanoparticles. To explore this vast parameter space and predict the mesophases of the hybrids, we have developed a mean field theory for mixtures of soft, flexible chains and hard spheres. Applied to diblock-nanoparticle mixtures, the theory predicts ordered phases where particles and diblocks self assemble into spatially periodic structures. The method can be applied to other copolymer-particle mixtures and can be used to design novel composite architectures. PMID- 11431563 TI - Diapiric flow at subduction zones: a recipe for rapid transport. AB - Recent geochemical studies of uranium-thorium series disequilibrium in rocks from subduction zones require magmas to be transported through the mantle from just above the subducting slab to the surface in as little as approximately 30,000 years. We present a series of laboratory experiments that investigate the characteristic time scales and flow patterns of the diapiric upwelling model of subduction zone magmatism. Results indicate that the interaction between buoyantly upwelling diapirs and subduction-induced flow in the mantle creates a network of low-density, low-viscosity conduits through which buoyant flow is rapid, yielding transport times commensurate with those indicated by uranium thorium studies. PMID- 11431564 TI - Evidence for a large-scale remnant of subducted lithosphere beneath Fiji. AB - We combine spatial variations of P- and S-wave speeds, 1000 fault plane solutions, and 6600 well-determined hypocenters to investigate the nature of subducted lithosphere and deep earthquakes beneath the Tonga back-arc. We show that perplexing patterns in seismicity and fault plane solutions can be accounted for by the juxtaposition of a steep-dipping Wadati-Benioff zone and a subhorizontal remnant of slab that is no longer attached to the actively subducting lithosphere. The detached slab may be from a previous episode of subduction along the fossil Vitiaz trench about 5 to 8 million years ago. The juxtaposition of slabs retains a large amount of subducted material in the transition zone of the mantle. Such a configuration, if common in the past, would allow the preservation of a primordial component in the lower mantle. PMID- 11431565 TI - A mite species that consists entirely of haploid females. AB - The dominance of the diploid state in higher organisms, with haploidy generally confined to the gametic phase, has led to the perception that diploidy is favored by selection. This view is highlighted by the fact that no known female organism within the Metazoa exists exclusively (or even for a prolonged period) in a haploid state. We used fluorescence microscopy and variation at nine microsatellite loci to show that the false spider mite, Brevipalpus phoenicis, consists of haploid female parthenogens. We show that this reproductive anomaly is caused by infection by an undescribed endosymbiotic bacterium, which results in feminization of haploid genetic males. PMID- 11431566 TI - Gene families from the Arabidopsis thaliana pollen coat proteome. AB - The pollen extracellular matrix contains proteins mediating species specificity and components needed for efficient pollination. We identified all proteins >10 kilodaltons in the Arabidopsis pollen coating and showed that most of the corresponding genes reside in two genomic clusters. One cluster encodes six lipases, whereas the other contains six lipid-binding oleosin genes, including GRP17, a gene that promotes efficient pollination. Individual oleosins exhibit extensive divergence between ecotypes, but the entire cluster remains intact. Analysis of the syntenic region in Brassica oleracea revealed even greater divergence, but a similar clustering of the genes. Such allelic flexibility may promote speciation in plants. PMID- 11431567 TI - Nitric oxide and the control of firefly flashing. AB - Bioluminescent flashing is essential for firefly reproduction, yet the specific molecular mechanisms that control light production are not well understood. We report that light production by fireflies can be stimulated by nitric oxide (NO) gas in the presence of oxygen and that NO scavengers block bioluminescence induced by the neurotransmitter octopamine. NO synthase is robustly expressed in the firefly lantern in cells interposed between nerve endings and the light producing photocytes. These results suggest that NO synthesis is a key determinant of flash control in fireflies. PMID- 11431568 TI - Contribution of aerobic photoheterotrophic bacteria to the carbon cycle in the ocean. AB - The vertical distribution of bacteriochlorophyll a, the numbers of infrared fluorescent cells, and the variable fluorescence signal at 880 nanometers wavelength, all indicate that photosynthetically competent anoxygenic phototrophic bacteria are abundant in the upper open ocean and comprise at least 11% of the total microbial community. These organisms are facultative photoheterotrophs, metabolizing organic carbon when available, but are capable of photosynthetic light utilization when organic carbon is scarce. They are globally distributed in the euphotic zone and represent a hitherto unrecognized component of the marine microbial community that appears to be critical to the cycling of both organic and inorganic carbon in the ocean. PMID- 11431569 TI - Nitrogen fixation by symbiotic and free-living spirochetes. AB - Spirochetes from termite hindguts and freshwater sediments possessed homologs of a nitrogenase gene (nifH) and exhibited nitrogenase activity, a previously unrecognized metabolic capability in spirochetes. Fixation of 15-dinitrogen was demonstrated with termite gut Treponema ZAS-9 and free-living Spirochaeta aurantia. Homologs of nifH were also present in human oral and bovine ruminal treponemes. Results implicate spirochetes in the nitrogen nutrition of termites, whose food is typically low in nitrogen, and in global nitrogen cycling. These results also proffer spirochetes as a likely origin of certain nifHs observed in termite guts and other environments that were not previously attributable to known microbes. PMID- 11431570 TI - Conditional restoration of hippocampal synaptic potentiation in Glur-A-deficient mice. AB - Plasticity of mature hippocampal CA1 synapses is dependent on l-alpha-amino-3 hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors containing the glutamate receptor A (GluR-A) subunit. In GluR-A-deficient mice, plasticity could be restored by controlled expression of green fluorescent protein (GFP)-tagged GluR A, which contributes to channel formation and displayed the developmental redistribution of AMPA receptors in CA1 pyramidal neurons. Long-term potentiation (LTP) induced by pairing or tetanic stimulation was rescued in adult GluR-A(-/-) mice when (GFP)GluR-A expression was constitutive or induced in already fully developed pyramidal cells. This shows that GluR-A-independent forms of synaptic plasticity can mediate the establishment of mature hippocampal circuits that are prebuilt to express GluR-A-dependent LTP. PMID- 11431571 TI - [Autoimmune peripheral neuropathies with anti-MAG antibodies and hematological disorders. Five cases]. AB - OBJECTIVE: The aim of this study was to report our experience with autoimmune neuropathies associated with hematological disorders and to describe their etiological and clinical polymorphism. PATIENTS AND METHODS: A retrospective study was conducted in five patients with autoimmune peripheral neuropathies with anti-MAG (myelin-associated glycoprotein) antibodies. RESULTS: Autoimmune neuropathies were associated with Waldenstrom's macroglobulinemia (n=2), Hodgkin disease (n=1), chronic lymphocytic leukemia (n=1) and idiopathic polyclonal B lymphoproliferation (n=1). Most of the patients had a sensorial polyneuropathy, predominant in the legs, exhibiting slow progress. Our patients showed a disappointing response to chemotherapy with stabilization or short response. PMID- 11431572 TI - Antigen inhalation as a triggering factor in systemic small-sized-vessel vasculitis. Four cases. AB - In order to identify small-sized-vessel vasculitis occurring immediately after massive inhalation of particles which could be considered antigenic, we performed a retrospective review of patients who consulted our Department of Internal Medicine between 1980 and 1998 and were diagnosed as having small-vessel vasculitis that developed immediately after massive exposure to inhaled particles. Four patients, 27 to 55 years old, presented with small-sized-vessel vasculitis (one with Wegener's granulomatosis and three with Churg-Strauss syndrome) which developed after massive inhalation of particles. Clinical manifestations occurred from a few hours to 10 days after inhalation of diesel fumes, cereal dust, flour or cereal dust and/or pigeon droppings. Three patients remain disease-free 8, 9 and 18 years after discontinuing treatment. One patient remains well after 3 years on low-dose corticosteroids and cyclophosphamide therapy. Our data suggest that primary antigens can cause small-sized-vessel vasculitides. Identifying such etiological factors could improve our understanding of the pathogenesis of systemic vasculitides and prevent relapses due to similar antigenic exposure. PMID- 11431574 TI - [Metabolism of atherogenic lipoproteins]. AB - Early development of atherosclerosis results from a complex multifactorial process where lipoprotein anomalies play a predominant role. The metabolism of lipoproteins is regulated by numerous reactions between the structural components of the lipoproteins and the receptors and/or enzymes with which they interact. Among the well-characterized anomalies, the elevation of small and dense LDL and/or the diminution of HDL levels are in first line of the factors involved in the formation of atheromatous plaques. LDL play a direct role by penetrating the intima of the arteries and HDL play an reverse transport of cholesterol from cells to the liver. There has been a long debate concerning risk related to triglyceride rich lipoproteins (TGRL), and more particularly the increase in VLDL. However, a large number of studies have demonstrated that these ephemeral lipoproteins can acquire major atherogenic potential when their level is increased and/or they are associated with perturbed metabolism leading to an accumulation of remnants. Current investigation methods have shown that LDL and HDL-cholesterol levels are excellent markers of LDL and HDL concentrations. Inversely, triglyceride levels provide little information concerning the nature of the elevated TGRL and fasting hypertriglyceridemia, even if moderate, should therefore be considered as a warning sign of persistent atherogeneous remnants in the fasting state. PMID- 11431575 TI - [Postprandial hyperlipidemia, small and dense LDL, HDL sub-fractions]. AB - The association of postprandial hyperlipidemia, small and dense LDL particles and low HDL cholesterol levels is a major cardiovascular risk factor, highly prevalent in insulin resistant and diabetic patients. Several recent epidemiological studies have demonstrated that an abnormal increase in the postprandial triglyceride levels is an independent cardiovascular risk factor, independent from fasting triglyceride levels. A decreased clearance of postprandial triglycerides is related to an abnormal intravascular lipoprotein metabolism, most of the time secondary to an insulin resistant state and genetic factors. This abnormal lipoprotein metabolism also induces a redistribution of LDL particles towards small and dense particles and a decrease in the HDL cholesterol levels. Small, dense LDL are associated with a 3 fold increase in the risk of ischemic heart disease, but does not remain a significant risk factor after adjustment for triglyceride levels. Decreased HDL cholesterol and apolipoprotein A-I levels are strong cardiovascular risk factors, which does not seem to be better assessed with the assay of various HDL sub-fractions (HDL(2) et HDL(3), LpA-I et LpA-I: A-II.). PMID- 11431576 TI - [New non-lipidic biological markers of atherosclerosis]. AB - Recent research indicates that both inflammation and infection of coronary arteries are intimately involved in the development and the progression of atherosclerosis. Pathophysiological mechanisms of formation of the atheromatous plaque are complex, resulting from the activation of endothelial cells, monocytes and foam cells, and smooth muscle cells. Numerous molecules are involved in these mechanisms, including adhesion molecules, pro-inflammatory and anti-inflammatory cytokines, chemokines, extracellular matrix remodelling enzymes, coagulation and fibrinolysis factors and infectious agents. Studies have suggested a potential role for these molecules as markers of the development of coronary artery disease and of adverse cardiac-related outcomes in patients with known coronary artery syndromes, in combination with independent risk factors such as homocysteine. This review presents recent data that could contribute to use of these markers for the diagnosis and the evolution of atherosclerosis. in clinical practice. PMID- 11431577 TI - [Revision of old risk factors: triglycerides]. AB - Serum triglycerides are transported by low-density lipoproteins: chylomicrons and VLDL. In the blood compartment, these lipoproteins undergo the action of lipoprotein lipase and are transformed into more dense particles captured by the liver. Several epidemiology studies have evidenced a strong relationship between serum triglyceride level and ischemic cardiovascular risk. This relationship tapers of or disappears after corrections for age, weight and HDL level in multivariate analysis. A recent meta-analysis demonstrated that the relative risk of cardiovascular events reached 1.14 in men and 1.32 in women, independently of confounding factors. Thus at the population scale, the impact of triglycerides on cardiovascular risk is weak. Certain forms are associated with high cardiovascular risk. For example, agarose gel electrophoresis can be used to identify a form of hypertriglyceridemia characterized by an accumulation of two populations of VLDL (double pre-ss). The prevalence of clinically patent vascular manifestations is 4 times greater in patients with this form of dyslipidemia than in subjects with hypertriglyceridemia without the double pre-ss. These observations point out the need for developing discriminating diagnostic methods to identify hyptriglyceridemic individuals with high cardiovascular risk. PMID- 11431578 TI - [Secondary prevention of coronary artery disease with lipid-lowering drugs]. AB - Before the statins era, this was one of the most debated questions in the field of cardiology: although hypolipidemic drugs (mainly fibrates) decreased the cardiovascular morbidity and mortality, their influence on total mortality was not significant. The 4S study demonstrated the efficacy of a statin on cardiovascular morbidity and mortality and on total mortality. This was confirmed by the CARE and the LIPID trials. Statins should be prescribed to all coronary patients, whatever the type of coronary disease, the age (we do not know yet for people older than 80), the gender, the basal cholesterol level (maybe statins are not efficacious when it is<2g/l); cholesterol level has to be lowered below 2g/l. Two trials of drugs increasing the HDL cholesterol level were recently published. Unfortunately their results are contradictory. Statins should be the first level drugs. Maybe it is useful to decrease the triglycerides level and to increase the HDL cholesterol level. The association of a statin and a fibrate is reserved to specialized centers. We should not forget the usefulness of diet. We must also take the other risk factors into account. PMID- 11431579 TI - [Does statin therapy reduce the risk of stroke? A meta-analysis]. AB - Large scale clinical trials have clearly demonstrated that the HMG-CoA reductase inhibitors (statins) reduce cardiovascular mortality by about 30%. The specific benefit on stroke prevention remains however to be determined. We reviewed all controlled clinical trials comparing statins versus placebo in primary and secondary prevention of cardiovascular disease. We identified 13 studies including 4S, CARE, WOSCOPS and LIPID. More than 32000 patients were randomized. The meta-analysis was performed using relative risk as treatment effect parameter. Statin treatment induced a significant relative risk reduction (RRR) of 24% (95% CI [12%-34%]) for stroke (2.1% vs 2.8%). RRR achieved 25% (95% CI [17%-32%]) for cardiovascular mortality and 34% (95% CI [30%-38%]) for myocardial infarction, without heterogeneity between trials. Stroke was reduced by 25% in secondary prevention, and by 15% in primary prevention, without significant heterogeneity between them. RRR of stroke was similar with pravastatin (RRR=0.79, p=0.0038) and with simvastatin (RRR=0.71, p=0.049). The effect model analysis (relationship between annual incidence of events in treated group versus placebo group in each trial) showed that RRR was constant whatever the baseline risk. These results are in favor of a preventive efficacy of statin treatment against stroke in middle aged patients with coronary heart disease. Complementary information will be needed to clarify the mechanism of this beneficial effect and to demonstrate statin efficacy in a population with a higher risk of stroke such as the elderly. PMID- 11431580 TI - [Role of diet in cardiovascular prevention]. AB - Several dietary components increase the risk of ischemic cardiovascular disease: excess dietary cholesterol and fats (especially saturated fats with low mono- and polyunsaturated fatty acid content), deficiency of antioxidants, fibers. These dietary factors modify the levels and the physicochemical structure of plasma lipoproteins and consequently their metabolic activities. Some of them also modify platelet aggregation and endothelial function. To reduce the cardiovascular risk, different nutritional schemes have been proposed and applied in prevention studies. Mediterranean-type diet, or traditional Asian food provide a very favorable cost/effectiveness ratio. These diets must be associated with the usual measures for improving lifestyle (increased physical activity, reduction of smoking) and with drug treatment of major risk factors (dyslipidemia, hypertension, diabetes). PMID- 11431581 TI - [Dietary recommendations for dyslipidemic individuals]. AB - Diet prescription is a fundamental first-line element in the management of patients with dyslipidemia. Weight loss should be a primary goal for all overweight patients. A 5 to 10% weight loss is often sufficient to obtain a significant improvement in lipid levels. Dietary fat should not provide more than 30% of total calorie intake. Saturated fatty acids should not exceed 7% and polyunsaturated fatty acids should also be limited to 7 to 10%. For monounsaturated fatty acids the dietary allowance can be a bit wider, up to 15% of total calorie intake. Carbohydrates, particularly complex carbohydrates, should replace the fat calories. Dietary fiber (more than 20g/d) as well as soy protein or phyto-sterols can be helpful in reducing LDL-cholesterol by about 10%. Consumption of fruits and vegetables should also be encouraged because they provide antioxidants that have effects on other cardiovascular risk factors. Long term education is needed to encourage the patient to comply with this type of diet. Psycho-behavioral strategies can be useful here. PMID- 11431582 TI - [Acquired factor VIIIc inhibitors. Two cases and a review of the literature]. PMID- 11431583 TI - Prolactin levels in Behcet's disease: no correlation with disease manifestations and activity. AB - OBJECTIVE: To assess prolactin level and its possible role in the pathogenesis and disease expression of patients with Behcet's disease (BD). MATERIALS AND METHODS: Twenty-eight patients with the diagnosis of Behcet's disease were selected. They all fulfilled the international criteria for BD. The control group consisted of 17 males and 6 females. Patients are divided into subgroups according to the clinical and paraclinical characteristics such as disease duration, ocular, articular and neurological involvement. Serum prolactin was measured by ELISA using Merck Kits on a Maggia 7000 Analyser. The normal range in males was<17ng/ml in males and<18ng/ml in females. The mean PRL level in each group and subgroup was calculated and compared with Student's t test. RESULTS: The mean prolactin level in the BD group (mean=13.76, SD=6.82), was higher than in the control group (mean=10.13, SD=5.46) with no statistically significant difference. The mean prolactin levels in all subgroups of patients with BD were higher than normal, but no statistically significant difference was shown between these subgroups. CONCLUSION: Hyperprolactinemia occurred in a small number of patients with BD and its significance remained unclear. Serum PRL level did not correlate with disease manifestations and activity. PMID- 11431584 TI - [Polyarteritis nodosa manifesting as tibial periostitis]. PMID- 11431585 TI - [Agranulocytosis and epileptic seizure during prolonged ceftriaxone treatment]. PMID- 11431586 TI - Bronchial brush: an unusual cause of bronchiectasis. PMID- 11431587 TI - [Smell dysfunction in nasal and paranasal sinus disease: a review of the literature (I)]. AB - Substantial advances have been achieved during the last decade in our understanding of the biological bases of the sense of smell, as well as in the clinical identification, diagnosis, and management of dysosmia. Nasal obstruction can result from inflammatory, neoplastic, traumatic, and developmental alterations within the nasal cavity. All these processes, if they result in bilateral restriction of airflow to the olfactory neuroepithelium, presumably alter the ability to smell. Rhinitis, nasal polyposis, and rhinosinusitis are accompanied by decreased ability to smell. Benign and malignant neoplasms can obstruct the nasal chamber and thereby alter airflow to the olfactory receptors without damaging the olfactory neuroepithelium. The purpose of this synthesis is to provide an advanced review of the literature in order to describe the basis of smell alterations in nasal polyposis and chronic rhinosinusitis. PMID- 11431588 TI - [Smell dysfunction in nasal and paranasal sinus disease: a review of the literature (II)]. AB - Substantial advances have been achieved during the last decade in our understanding of the biological bases of the sense of smell, as well as in the clinical identification, diagnosis, and management of dysosmia. Nasal obstruction can result from inflammatory, neoplastic, traumatic, and developmental alterations within the nasal cavity. All these processes, if they result in bilateral restriction of airflow to the olfactory neuroepithelium, presumably alter the ability to smell. Rhinitis, nasal polyposis, and rhinosinusitis are accompanied by decreased ability to smell. Benign and malignant neoplasms can obstruct the nasal chamber and thereby alter airflow to the olfactory receptors without damaging the olfactory neuroepithelium. The purpose of this synthesis is to provide an advanced review of the literature in order to describe the basis of smell alterations in chronic rhinitis, nasal malformations and tumors. PMID- 11431589 TI - [Adenocarcinoma of the ethmoid sinuses: results of a retrospective study in Rouen]. AB - Ethmoid adenocarcinoma is a rare tumour of the ethmoidal sinuses. The authors report on the clinical features, treatment and follow-up results in 19 cases. Risk factors were those regularly encountered. Delay to diagnosis was long due to the nonspecific clinical features and course. Nasal endoscopy was essential for follow-up. Computed tomography and magnetic resonance imaging were also required to assess tumour spread. Our results suggest that radiotherapy following surgery should be preferred. Survival rate is generally low for this type of tumor. We had 77% survival at 5 years. PMID- 11431590 TI - [Supraglottic laryngectomy with arytenoidectomy]. AB - Relearning to swallow is frequently difficult after supraglottic laryngectomy requiring arytenoid cartilage resection. We propose a surgical procedure in which a local flap is used to close the pharyngeal defect without approximating the laryngeal remnants and the base of tongue. The procedure opens the median raphe and cuts the hyoid bone along the midline. The strap muscles, the perichondrium from the thyroid cartilage and thyroid lobe on the ispilateral side to the tumor are retracted laterally to be used to close the mucosal defect. Sixty-eight patients with T1-T3 carcinomas of the laryngeal margin with extension to an arytenoid in all cases and limited extension to the medial wall and/or anterior angle pyriform fossa in 33 underwent this surgical procedure. Only 5 patients had local recurrence and 8 a lymph node recurrence. Visceral metastases occurred in 21 patients (33%) and second primary tumors were diagnosed at the time of surgery or during follow-up in 19 patients (28%). Three and 5-year actuarial survival rates were 57 and 51%, respectively. Despite post operative radiotherapy, functional success was obtained in 50 patients (75%). This technique provided good tumor control and a high rate of satisfactory functional results in patients with tumors of lateral margin extended to one arytenoid. PMID- 11431591 TI - [Inner ear and subarachnoid spaces: relations and diseases]. AB - The purpose of this article is to describe, with 5 clinical cases, the physiological communications between the inner ear and the subarachnoid spaces (SAS) and present the imaging features with regard to. Therefore we briefly illustrate abnormal communications between SAS and perilymphatic fluids in certain cochlear and internal acoustic meatus (IAM) malformations and their consequences. Imaging features may depict diffusion pathway of bacterial meningitis to membranous labyrinth via the cochlear aqueduct or via the IAM. Rarely, in some patients referred for cochleovestibular symptoms, imaging features may display skull base tumors involving the area of cochlear or vestibular aqueduct aperture. Therefore, in patients referred for cochleovestibular symptoms, MR and CT study should carefully scrutinise not only the IAM but also the aperture of the cochlear and the vestibular aqueducts and the cerebellopontine meninges. PMID- 11431592 TI - [Labyrinthine fistulae and cholesteatoma]. AB - Labyrinthine fistula is a potentially serious complication of cholesteatoma. During or after surgery, cholesteatoma can induce partial or total destruction of cochleovestibular functions. We operated 38 patients from 1983 to 1996. The site of the fistula was the external semi-circular canal in 90% of the cases. The facial nerve canal was eroded in 66% of the cases. We performed 11 CT scans; only 7 evidenced the fistula. We removed the matrix of the cholesteatoma during the initial surgery in 35 cases and in 3 left the fistula in situ for subsequent excision at a second operation. Postoperative hearing loss compared with the preoperative situation was observed in 66% of the patients. Deafness was observed in 4 ears (11%). Hearing improved after surgery in 23% of the patients. We consider that a closed technique with immediate removal of the cholesteatoma matrix is indicated for most fistulae but that second-intention resection (combined approach tympanoplasty) is the better choice when the fistula is wide and the ear is infected. In some cases (old patient, one functional ear, better ear) an open technique may be preferred without risk for the cochleovestibular functions. PMID- 11431593 TI - [Pharyngeal Wegener's disease: a case report]. AB - We report a case of Wegener's disease with an exceptional 10-year course before diagnosis subsequent to the development of severe ENT features. Mucosal ulcerations involving the mouth and the pharynx were highly extensive and resolved completely, but temporarily, with cyclophosphamide and corticosteroid therapy. PMID- 11431594 TI - [Surgical treatment for squamous-cell carcinoma of the posterior pharyngeal wall: reconstruction with a free forearm flap]. AB - Squamous-cell carcinoma of the posterior pharyngeal wall is a rare finding. Surgical treatment with preservation of laryngeal function is usually associated with relatively poor functional outcome. We report the cases of 5 patients treated by pharyngectomy with preservation of the laryngeal function. The posterior pharyngeal wall was reconstructed with a free forearm flap. This reconstruction method provided satisfactory functional results. Swallowing disorders resolved within 2 to 6 weeks following the reintroduction of oral food intake. Mean follow-up of nearly 3 years did not evidence any local tumor recurrence. PMID- 11431595 TI - Thiazolidinediones and liver toxicity. AB - Thiazolidinediones or glitazones specifically target insulin resistance. They have proven efficacy for reducing plasma glucose levels of type 2 diabetic patients treated with diet alone, sulphonylureas, metformin or insulin. In addition, they may be associated to some improvement of cardiovascular risk profile. However, troglitazone, the first compound approved by the FDA in the US, proved to be hepatotoxic and was withdrawn from the market after the report of several dozens of deaths or cases of severe hepatic failure requiring liver transplantation. It remains unclear whether or not hepatotoxicity is a class effect or is related to the unique tocopherol side chain of troglitazone. Rosiglitazone and pioglitazone, two other glitazones, appear to have similar efficacy on blood glucose control of type 2 diabetic patients as compared to troglitazone. In controlled clinical trials, the incidence of significant increases in liver enzyme levels (ALT) was similar with rosiglitazone or pioglitazone as compared to placebo, whereas troglitazone was associated with a threefold greater incidence. In contrast to the numerous case reports of acute liver failure in patients receiving troglitzone, only two cases of severe reversible liver failure have been reported in patients treated with rosiglitazone, with a causal relationship remaining uncertain. Furthermore, no single case of severe hepatotoxicity has been reported yet with pioglitazone. While regular monitoring of liver enzymes is still recommended and more long-term data are desirable, current clinical evidence supports the conclusion that rosiglitazone and pioglitazone do not share the hepatotoxic profile of troglitazone. PMID- 11431596 TI - Increased levels of soluble Fas in serum from diabetic patients with neuropathy. AB - OBJECTIVE: The aim of this study was to investigate circulating soluble Fas (sFas) and Fas ligand (sFasL), two transmembrane glycoproteins involved in apoptosis, in the serum of diabetic patients. MATERIAL AND METHODS: We assessed sFas and sFasL serum levels in normal controls (n=15), and in both 42 diabetic patients without complications, or with predominant retinopathy or neuropathy, using sFas and sFasL specific ELISA method. RESULTS: sFasL serum levels were less than 0.1 ng/ml in normal controls and in each group of diabetic patients. In diabetic patients with a predominant neuropathy, sFas serum levels were significantly increased not only when compared with normal controls (13.5 +/- 3.6 ng/ml vs 7.1 +/- 1.1 ng/ml, p<0.001), but also when compared with patients without complications (vs 9.1 +/- 1.8 ng/ml, p<0.001) or with a predominant retinopathy (vs 8.7 +/- 1.9 ng/ml, p<0.001). CONCLUSIONS: These preliminary data suggest that a dysregulation of the Fas system in peripheral neuronal cells may be involved in the increase of sFas observed in diabetic patients with neuropathy. PMID- 11431597 TI - A randomized study comparing blood glucose control and risk of severe hypoglycemia achieved by non-programmable versus programmable external insulin pumps. AB - OBJECTIVE: To compare a non-programmable and a programmable insulin external pump using regular insulin on glycemic stability, the risk of severe hypoglycemia and metabolic control in type 1 diabetic patients. MATERIAL AND METHODS: Ten type 1 diabetic patients were involved in a randomized, crossover study comparing two periods of 3 months with continuous subcutaneous insulin infusion (CSII) either with a non-programmable insulin pump or a programmable insulin pump. Comparisons were made among mean blood glucose values before and after meals, at bedtime and at 2: 00 a.m.; the risk of severe hypoglycemia assessed by the low blood glucose index (LBGI); and HbA1c. RESULTS: Mean average blood glucose (BG) measurements were significantly lower with the programmable in comparison with the non programmable insulin pump (respectively 157+/-78 vs. 165+/-79, p=0.034). While postprandial values for BG were not different between the two pumps, the use of the programmable pump resulted in a significant decrease in mean preprandial BG levels (140+/-68 vs. 150+/-73 mg/dl p=0.039). Conversely mean BG level was lower at 2 a.m. with the non-prgrammable pump (125+/-81 vs. 134 +/-93 mg/dl, p=0.02) but with a higher incidence of hypoglycemia. Mean LBGI was comparable with the two pumps (3.1+/-8.6 vs. 2.8+/-6.9, p=0.1). There was a 0.2% decrease in HbA1c during the programmable pump period that did not reach statistical significance (p=0.37). CONCLUSIONS: The present study suggests that programmable external insulin pumps, although more complex and more expensive than non-programmable insulin pumps, significantly reduce fasting glycemia during the day without increasing the risk of severe hypoglycemia and are safer during the night. PMID- 11431598 TI - Erectile dysfunction and lower androgenicity in type 1 diabetic patients. AB - OBJECTIVE: To analyse the clinical characteristics and relevant hormonal profile in type 1 diabetic patients with and without ED. MATERIAL AND METHODS: Fifty one type 1 diabetic patients were studied. ED was assessed by direct interview. Chronic diabetic complications, smoking and alcohol status as well as current use of medications were recorded. Hormonal profile consisted of plasma LH, FSH, prolactin, androstenedione (Delta(4)), dehydroepiandrosterone (DHEA), DHEA sulfate (DHEA-S), free testosterone (FT), estradiol (E(2)), sex hormone binding globulin (SHBG), dihydrotestosterone (DHT), cortisol, TSH and free thyroxine (FT(4)). RESULTS: ED was present in 24 patients (47%) (group 1), who were older (P<0.001), had a longer diabetes duration (P<0.001) and a higher systolic blood pressure (P=0.017) when compared to the subjects who did not complain (group 2). ED was positively correlated to all diabetes-related complications (P<0.02). Antidepressive drug(s) were more frequent in group 1 (P=0.007), as well as prokinetics (P=0.043) and ACE-inhibitors (P=0.010). HbA(1)c was comparable. Patients with ED had lower levels of Delta(4) (P=0.003), DHEA (P<0.001), DHEA-S (P=0.002), FT (P=0.08) while SHBG (P=0.010) and LH (P=0.022) were higher compared to group 2. Multiple logistic regression analysis showed an independent association of ED with Delta(4) (P=0.016), DHEA-S (P=0.037), SHBG (P=0.001) and insulin dose (P=0.025). There was no significant difference for all other measured hormones. CONCLUSION: ED is impressively prevalent in type 1 diabetes and is associated with age, diabetes duration, chronic complications and decreased androgens. PMID- 11431599 TI - Heterogeneity of fetal growth in type 1 diabetic pregnancy. AB - OBJECTIVE: To investigate the frequency of macrosomia in an homogeneous cohort of type 1 diabetic mothers and to analyze the influence of maternal factors and glycemic control on the incidence of fetal macrosomia. MATERIAL AND METHODS: Fifty-five consecutive type 1 diabetic first-pregnancies were prospectively studied. Macrosomia was defined by a ponderal index above the 90(th) percentile. Venous cord blood levels of insulin, C peptide and leptin were measured at delivery. The influence of HbA1c levels and other maternal variables on the occurrence of macrosomia and on the ponderal index was assessed using a stepwise regression logistic model. RESULTS: The mean (+/- SD) birth weight was 3482 (+/- 497) g at 37.4 +/- 1.0 weeks gestation. Macrosomia occurred in 29 cases (53.7%). Fetal insulin, C peptide and leptin levels were significantly higher in macrosomic than in non macrosomic infants. Maternal age, duration of diabetes, pregravid body mass index, parity, weight gain during pregnancy, presence of a microangiopathy, nephropathy, smoking habits, gestational hypertension or preeclampsia, and HbA1c levels throughout pregnancy did not differed between mothers of macrosomic and non macrosomic infants. In the stepwise analysis none of these covariates was explanatory of the ponderal index. CONCLUSIONS: The frequency of macrosomia remains very high in infants of type 1 diabetic mothers despite a reasonable degree of glycemic control. The variability of the fetal growth response to mild hyperglycemia prompts for the identification of other factors involved in the modulation of fetal growth. PMID- 11431600 TI - Prevalence of type 2 diabetes and impaired fasting glucose in the middle-aged population of three French regions - The MONICA study 1995-97. AB - OBJECTIVES: The aim of this study was to determine the prevalence of type 2 diabetes and impaired fasting glucose (IFG) in a population-based sample of 3 508 subjects, aged 35-64 years, participating in the French MONICA population survey from 1995 to 1997 in three French regions: the Urban Community of Lille, the Bas Rhin and the Haute-Garonne. MATERIAL AND METHODS: Previously diagnosed type 2 diabetes is defined by the current use of oral hypoglycaemic treatment and newly diagnosed subjects by fasting plasma glucose (FPG) > or =7.0 mmol/L according to the ADA 1997 recommendations. IFG was determined by 6.1< or =FPG< or =6.9 mmol/L. Adjusted prevalences are calculated according to the French 1990 census data. RESULTS: Type 2 diabetes adjusted prevalence is 5.1% [4.1-6.1] in women and 7.3% [6.1-8.4] in men while IFG adjusted prevalence is 5.2% [4.2-6.2] and 11.8% [10.3 13.4] respectively. Prevalences of type 2 diabetes and IFG are both significantly higher in men than in women. This trend appears in any age group for IFG, but is only observed in 55-64 year-old subjects for type 2 diabetes. The reduction of the FPG threshold to screen diabetes mellitus from 7.8 to 7.0 mmol/L according to the ADA recommendations results in a 2.2-fold increase in the number of newly diagnosed diabetic subjects, screened by one FPG measurement, in our population based sample. CONCLUSIONS: The MONICA population survey confirms that type 2 diabetes represents a major health care problem in France and underlines the influence of gender on the prevalence of both type 2 diabetes and IFG in the French middle-aged population. PMID- 11431601 TI - Gastric emptying is accelerated in obese type 2 diabetic patients without autonomic neuropathy. AB - OBJECTIVE: To clarify the impact of type 2 diabetes mellitus on the gastric emptying rate. MATERIAL AND METHODS: Using a double-isotope scintigraphic technique, we assessed the gastric emptying of a standard liquid-solid meal in 13 obese type 2 diabetic patients without autonomic neuropathy (age: 47.4 +/- 8.6 yr, body mass index: 33.9 +/- 4.8 kg/m(2), glycaemia: 9.1 +/- 2.6 mmol/l) and in 7 controls with similar sex ratio, age, BMI and body fat distribution. RESULTS: The half gastric emptying time for the liquid phase was not significantly different between diabetic patients and controls (respectively: 52.7 +/- 14.5 min and 63.1 +/- 15.2 min). However, the half gastric emptying time for the solid phase was significantly shortened in diabetic patients versus controls (respectively 88.8 +/- 23.2 min in diabetic patients and 113.6 +/- 26.9 min in controls; p<0.04). Furthermore, a negative relationship was highlighted between the half gastric emptying time for the solid phase and basal glycaemia (r=-0.65, p<0.02) in diabetic patients. No significant relationship was found between gastric emptying parameters and cardiac autonomic nerve function, insulin or gastrin levels. CONCLUSION: Solid gastric emptying is accelerated in obese type 2 diabetic patients without patent autonomic neuropathy when compared to obese non diabetic patients. PMID- 11431602 TI - What may be gained from standard photocoagulation during early worsening of diabetic retinopathy? An observational study in type-1 diabetic patients after tightening of glycaemic control. AB - OBJECTIVE: To assess the outcome of laser photocoagulation treatment for rapidly progressing diabetic retinopathy, socalled early worsening, subsequent to a rapid improvement of glycemic control. For the purpose of this study, early worsening was defined as any incidence or progression of retinopathy that followed a reduction in HbA1c by > 2% within 6 months. MATERIAL AND METHODS: Retrospective observational study in type-1 diabetic patients in a university diabetes center. PATIENTS: 23 patients with early worsening were identfied during a 16-year period, with a mean age of 25 years, duration of diabetes of 12 years, and glycated hemoglobin HbA1c of 12.4%; retinopathy was absent or mild nonproliferative at baseline. Focal, and/or panretinal laser coagulation was performed according to standard ETDRS criteria. Retinal pathology and visual acuity was followed-up for 12-120 months. RESULTS: Improving metabolic control induced mild non-proliferative retinopathy without macular edema in 4 patients, which regressed without treatment. In 19 patients, symptomatic diabetic maculopathy developed with macular edema, resolving by focal coagulation in 3 patients. Of the remaining 16 patients, 14 developed proliferative retinopathy (7 of whom despite focal, grid or scatter coagulation pretreatment), and were treated by full panretinal coagulation. In 7 of the 14 patients with proliferative retinopathy, vitreous hemorrhages occurred requiring pars plana vitrectomy. Proteinuria, polyneuropathy, and impaired vision prior to laser treatment were indicative of poor prognosis. Visual acuity > 0.3 in at least one eye was preserved in 22 of the 23 patients. CONCLUSIONS: In patients with type-1 diabetes mellitus and early worsening of diabetic retinopathy, the benefit of standard laser photocoagulation was limited, and particularly in the presence of symptomatic macular edema. PMID- 11431603 TI - Six month administration of gelified intranasal insulin in 16 type 1 diabetic patients under multiple injections: efficacy vs subcutaneous injections and local tolerance. AB - OBJECTIVE: Nasal insulin administration is a potential route for intensive insulin management, less invasive and more rapid than subcutaneous injections. Previous studies have shown poor bioavailability (less than 15%) with nasal insulin administration with various absorption enhancers. The aim of the study was to evaluate in type 1 diabetic patients, the metabolic efficacy and local tolerance of a new gelified sprayed nasal insulin containing glychocolate and methylcellulose as absorption promoters. MATERIAL AND METHODS: The study was conducted in 16 type 1 diabetic patients (HbA1c 8.6+/-0.2%) in a cross-over trial including 2 six month randomized periods: a) NPH twice daily + 3 pre-prandial nasal insulin doses + nasal supplementation in case of unexpected hyperglycaemia; b) NPH twice daily + 3 pre-prandial regular insulin injections. End points were HbA1c levels, hypoglycaemic episodes and tolerance evaluated at month 0, 2, 6 and 8 on clinical symptoms and objective nasal assessments. RESULTS: Four patients were withdrawn because of nasal burning (3 cases) and persistent sinusitis (1 case), and one patient had purulent sinusitis at the month 6 examination. At month 6, HbA1c levels were comparable (8.3 +/- 0.1 vs 8.6 +/- 0.1%, m +/- SEM, NS) for nasal and subcutaneous period respectively. The number of hypoglycaemic events was identical during the 2 periods (88 episodes). Nasal tolerance with the gelified form was better than with the already reported lyophilized form but, when present, symptoms were more marked, suggesting a potentiating additional role of methylcellulose excipient on nasal intolerance. CONCLUSIONS: 1) Gelified nasal insulin is as efficient as subcutaneous regular insulin in type 1 diabetic patients. 2) Other galenic forms should be investigated to improve nasal tolerance and bioavailability. PMID- 11431604 TI - Left ventricular mass and systolic function in African diabetic patients: association with microalbuminuria. AB - OBJECTIVE: To assess echocardiographic evidence of cardiomyopathy and its association with microalbuminuria in type 2 normotensive non-proteinuric diabetic patients. MATERIAL AND METHODS: Forty consecutive normotensive non-proteinuric type 2 diabetic patients were studied. Body mass index, blood pressure, urinary albumin excretion, ECG at rest and after exercise, left ventricular mass, and shortening fraction using two-dimensional and M-mode echocardiography were measured in every patient. RESULTS: Among the 40 patients studied, 17 (42.5%) presented with microalbuminuria, 16 (40.0%) with left ventricular hypertrophy, 22 (55.0%) with systolic dysfunction and 3 (7.5%) with ECG changes compatible with cardiac ischaemia. No significant difference existed between normoalbuminuric and microalbuminuric patients for age, known duration of diabetes, body mass index, systolic and diastolic blood pressure. Ventricular mass correlated to urinary albumin excretion rate (r=0.34; p=0.04) and shortening fraction to diastolic blood pressure (r = - 0.40; p=0.01). CONCLUSION: Left ventricular structure and function might be altered in African type 2 diabetic patients in the absence of hypertension, and microalbuminuria may be an early biochemical marker of these abnormalities. PMID- 11431605 TI - Erythropoietin-dependent anaemia: a possible complication of diabetic neuropathy. AB - We report the case of a 52-year-old woman with long-term type 1 diabetes mellitus, complicated with proliferative retinopathy, autonomic neuropathy and microalbuminuria and moderate renal failure. A normochromic, normocytic are generative anaemia had been diagnosed for three years. Clinical and biological investigations for the aetiology of anaemia remained normal or negative. Anaemia was associated with a concentration of erythropoietin (EPO) in the normal range, but inappropriately low regarding anaemia. Treatment with recombinant EPO induced a rapid increase in haemoglobin level and improved the patient's quality of life. The role of diabetic neuropathy in the genesis of anaemia, in conjunction with a modest renal impairment is discussed. PMID- 11431606 TI - [Insulin doses adaptation with insulin Lys-pro in external pump]. PMID- 11431607 TI - [Dietary assessment in current clinical practice: how to conciliate rapidity, simplicity and reliability?]. AB - Dietary interviews and food diaries are traditionally used for nutritional assessments. In clinical practice, these methods are time consuming, require high training, and thus remain poorly used. Furthermore, the results are frequently impaired by the underreporting phenomenon which can be due either to underrecording (failure to record what is eaten) or to undereating (volontary food restriction during the assessment period). These difficulties can be overcome by using rapid questionnaires based on 2 principles: 1) underreporting is less for proteins than for other macronutrients; 2) in developed countries, calories from proteins are relatively stable and contribute approximately to one sixth of the total daily energy intake. Estimations given by the rapid questionnaire lead to less misleading results than those provided by 7 day-food records. On the other hand, the rapid questionnaire gives an estimate of specific dietary behaviors such as nibbling, festive meals and consumption of salted entrees, sweet desserts and caloric beverages. In conclusion, helpful and simple recommendations for correcting main nutritional errors can be drawn from estimation of the above mentioned specific behaviors that correspond to a daily average of 500 kcalories. PMID- 11431608 TI - [Nuclear medicine in the diagnosis of diabetic foot osteomyelitis]. AB - The diagnosis of diabetic foot osteomyelitis is often difficult both clinically and radiologically with a delay in radiological sign occurrence and difficulties of imaging interpretation. Bone biopsy is known to be the diagnosis gold standard. However, if negative, the diagnosis of osteomyelitis cannot be excluded and this method is not harmless. An early diagnosis of osteomyelitis is necessary to start an antibiotic treatment in conjunction with conservative surgery. (99m)Tc-HMPAO labelled leucocyte scintigraphy performed in conjunction with bone scintigraphy significantly contributes to the diagnosis of osteomyelitis (sensitivity=100% and specificity > 95%). In case of osteomyelitis suspicion, after plain radiography, the (99m)Tc-MDP bone scintigraphy is the first step. If negative, osteomyelitis is unlikely. If positive, a (99m)Tc-HMPAO leucocyte scintigraphy should be performed in order to exclude or to confirm the diagnosis of bone infection. PMID- 11431609 TI - [Prevention of peritoneal adhesions]. AB - Peritoneal adhesions are a major problem for health and economy. An adequate and atraumatic surgical technique is essential in the prevention of peritoneal adhesions. Laparoscopic microsurgery should be developed and promoted. The following conclusions can be proposed about the treatments available Adequately designed clinical studies are too rare. Future studies are necessary to obtain information on pregnancy and pain as primary outcomes. Cost implications of adhesion prevention treatments also have to be addressed. PMID- 11431610 TI - [Vaginal hysterectomy in nulliparous women: indications and limitations]. AB - OBJECTIVE: The aim of our study was to determine whether nulliparity is a limiting factor for vaginal hysterectomy performed for benign lesions. PATIENTS AND METHODS: We conducted a prospective study in 1604 patients who underwent hysterectomy for a benign lesion between 1991 and 1998. We studied a subgroup of this population constituted by 128 nulliparous women, i.e. 8% of the patients. RESULTS: Vaginal hysterectomy was performed at first intention in 54.7% of the cases, after laparoscopic preparation in 14%, and by first intention laparotomy in 31.3% for each type of approach we compared: history of pelvic surgery, duration of the operation, final weight of the uterus, peroperative complications, association or not with adnexectomy in patients aged 45 years or older, frequency of a reduction procedure in the vaginal group and duration of hospitalization. CONCLUSION: Nulliparity is not a contraindication for vaginal hysterectomy which can be proposed as a first intention procedure, sometimes in association with laparoscopic preparation in case of prior pelvic surgery or concomitant adnexal disorders. PMID- 11431611 TI - [Mammary tuberculosis: a series of 14 cases]. AB - OBJECTIVES: We examined cases of mammary tuberculosis in order to determine the different modes of presentation and analyze diagnostic difficulties. METHODS: Retrospective analysis of 14 cases of mammary tuberculosis treated at the Ibn Rochd University Hospital in Casablanca over a 16 year period. RESULTS: Patient age ranged from 16 to 65 years. Contact with a contaminated person was recognized in 2 cases. The predominant clinical presentation was a tumor formation (12 cases) which sometimes simulated cancer. Axillary nodes were observed in 10 cases with 1 case of fistulization. Mammography suggested cancer in 3 cases. The diagnosis of mammary tuberculosis was based on pathology findings in 14 cases (2 biopsies, 12 peroperative specimens) and isolation of bacilli from pus in one case. Medical treatment was given. Surgery was associated as needed (abscess drainage, residual lesions). Except for one death due to tuberculous meningoencephalitis, clinical course was favorable with ad integrun breast recovery. DISCUSSION: Mammary tuberculosis is uncommon and often produces a pseudoneoplastic presentation. Pathology confirmation is required for diagnosis. PMID- 11431612 TI - [Regionalization of perinatal care in the Seine-Saint-Denis department of France]. AB - OBJECTIVE: To evaluate a policy designed to regionalize perinatal care in the Seine-Saint-Denis department of France. METHODS: The place of birth of every preterm infant (born before 33 weeks gestation) in 1998-1999 was compared with that for the period of 1989-1992. The 1989-1992 data came from a prenatal mortality study. For the 1998-1999 period, we used data from an area-based birth registry recording an experimental health certificate. RESULTS: In 1989-1992, 40% of live births before 33 weeks gestation took place in level I maternity units, 37.2% in level II maternity units, and 13.0% in level III maternity units. In 1998-1999, 5.4% took place in level I maternity units, 28.9% in level II maternity units and 65.1% in level III maternity units. The number of postnatal transfers of very preterm infants declined markedly. In 1998-1999, 109 pregnant women were transferred to a level III maternity hospital. This constituted 1.2% of the women who gave birth in Seine-Saint-Denis during this period. CONCLUSION: The policy to regionalize perinatal care and increase maternal transfers was well accepted and successfully implemented. The delivery of very preterm infants in maternity hospitals without neonatal units became a rare event. PMID- 11431613 TI - [Post-prandial plasma glucose test as screening tool for gestational diabetes: A prospective randomized trial]. AB - OBJECTIVE: To compare one hour postprandial glucose measurements with the one hour 50gm plasma glucose test as predictors of gestational diabetes. SUBJECTS AND METHODS: One hundred women at 24-28 weeks' gestation were prospectively randomized for both screening methods: one hour postprandial and one hour 50gm plasma glucose test with a one-week-interval between tests. A week later a formal 2-hour 75gm glucose tolerance test was done in each case. RESULTS: Of the 95 patients who completed the study 13 had gestational diabetes (13.6%). For a threshold of 7.1mmol/l the sensitivity of 1-hour postprandial plasma glucose screening test was 84.6% with a specificity of 87.8%. These values were respectively 84.6% and 81.7% for 1-hour 50 gm plasma glucose test screening test. A threshold of 7.7mmol/l yielded a sensitivity of 60.5% with a specificity of 91.4% for the 1-hour postprandial plasma glucose screening test (69.2% and 86.5% for 1-hour 50gm plasma glucose test) CONCLUSION: In our study the 1-hour postprandial plasma glucose screening test was as effective as the 1-hour 50 gm plasma glucose test screening test for gestational diabetes. PMID- 11431614 TI - [Towards simplified cesarean section?]. AB - In the past few years, many international publications have reported important improvements in the cesarean section procedure. Considering these improvements, M STARK suggests a new technique for the cesarean section which is called <> (=ML). To assess feasibility and efficiency of this technique the authors conducted a prospective randomised study for 4 months including all cesarean sections and compared the ML technique with the classical Pfannenstiel are. The following criteria were noted for each woman. Results showed the superiority of the M.L. technique especially with regard to the length of duration of the post-operative fever and the parietal blood collection, where difference is significant. Regarding the blood loss rate, the post-operation pain, the delay before gas and the duration of the hospitalisation period the M.L. technique seems to be better although the difference is not very significant statistically. Following this study and the international literature survey, authors conclude that the M. L technique is so much better than the pfannenstiel technique that it must be used throughout daily obstetrical practice. PMID- 11431615 TI - [Treatment of pain due to unwanted lactation with a homeopathic preparation given in the immediate post-partum period]. AB - Dopaminergic agonists, such as Parlodel((R)), are now widely used to inhibit lactation. However, some countries, such as the United States, no longer use these drugs in this indication because of their sometimes serious adverse effects. In this context, the authors tested a homeopathic treatment designed for parturients unable or not wanting to breastfeed. The APIS MELLIFICA 9 CH and BRYONIA 9 CH combination was chosen for its anti-inflammatory and analgesic effects. 71 patients were included in this double-blind placebo-controlled study. All received basic treatment comprising naproxen and fluid restriction. A significant improvement of lactation pain (main criterion of the study) was observed in parturients treated with homeopathy (p<0.02 on D2 and p<0.01 on D4). A similar effect (p<0.05 on D4) was observed for breast tension and spontaneous milk flow. No significant difference was observed for the other criteria of the study. The homeopathic combination studied was therefore effective on the pain of lactation and should be integrated into the therapeutic armamentarium. PMID- 11431616 TI - [Impact of systematically freezing embryos on ovum donation]. AB - In France, all embryos obtained after ovum donation have to be frozen. We present a brief background on this policy and expose our results, then discuss the rationale of such a policy in order to upgrade knowledge on the mechanism of vertical HIV transmission. PMID- 11431617 TI - [The debate on breach presentation]. PMID- 11431618 TI - [Medicine and the art of speaking. Honorary conference]. PMID- 11431619 TI - [Address of the chairman]. PMID- 11431621 TI - [A century of progress. From biology to antithrombotic agents]. PMID- 11431622 TI - [A century of progress. From macrocirculation to microcirculation]. PMID- 11431623 TI - [Introduction to vascular engineering]. AB - Vascular engineering developed through advancing knowledge of the physiological and pathological processes operating in cells and tissues subjected to environmental mechanical stress. We review briefly the more relevant cellular and general mechanism characteristic of vascular tissue examining the performances obtained with prosthetic materials (allografts, synthetic grafts) and current research and development of new vessel substitutes (biohybrids or biovessels). PMID- 11431624 TI - [Takayasu's disease associated with autoimmune thyroiditis and celiac disease. Clinical course and limitations of treatment]. AB - Takayasu's arteritis is a non-specific form of vasculitis involving the aorta, its main branches and pulmonary arteries. It is a rare disease in our country, contrasting with the high prevalence in Southeast Asia, Africa and South America. We discuss the course of the disease in our patient who was a young women who developed Takayasu's arteritis associated with autoimmune thyroiditis and malabsorption syndrome due to celiac sprue. Long-term immunosuppressive therapy contributed to stabilizing the associated diseases, but did not stop the progression of the vascular lesions. The main adverse outcome in our patient was the development of severe renovascular hypertension. Bilateral renal artery stenosis was treated by angioplasty with stent implantation. Based on literature reports, the association of Takayasu's disease with multiple autoimmune disorders is a rare event. However, it would appear that the arteritis was the limiting disease for prognosis in our patient. PMID- 11431625 TI - [Oral antivitamins K]. PMID- 11431626 TI - [Toxic skin reaction to clopidogrel]. PMID- 11431627 TI - [Unstable pelvic ring fracture: osteosynthesis of the posterior lesion by percutaneous sacroiliac fixation]. AB - PURPOSE OF THE STUDY: We report our experience with percutaneous sacroiliac screwing for unstable pelvic ring fracture. MATERIAL AND METHODS: Twenty unstable pelvic ring fractures with sacroiliac disjunction (9 cases), trans-sacral fracture (9 cases), or bilateral posterior fractures (5 cases) were managed with percutaneous sacroiliac fixation using 1 or 2 screws. There was also pubic disjunction in 7 cases, managed by osteosynthesis in 5 cases. No fixation of fractures of one or both of the obturator rings, present in 13 cases, was attempted. The posterior screwing was performed under fluoroscopic guidance in 15 cases and computed tomographic guidance in 5. RESULTS: There were no infectious or neurological complications. Radiographically, anatomic reduction of the posterior fracture was achieved in 10 cases. A vertical ascension persisted in 4 patients and a rotation in 6. Radiographic results were correlated with delay to surgery: correction of the vertical displacement was achieved in 80% of the patients operated before five days and in 55% of the others. Two patients experienced a secondary displacement. There was no cases of nonunion of either the anterior or posterior fracture. Follow-up was available for 14 patients (2 patients died and follow-up was too short for 4). The mean Majeed score was 92/100 and none of the patients experienced anterior pain. Three patients had moderate pain in the sacroiliac area. DISCUSSION: The ventral position appears to be best adapted for percutaneous sacroiliac screwing but can be contraindicated by concomitant lesions. We prefer fluoroscopic guidance for these emergency procedures. Two views, ascending and descending, appear sufficient. The better stability provided by two screws can be achieved under computed tomographic guidance without risk of neurovascular damage. CONCLUSION: Percutaneous sacroiliac screwing is a good management strategy for unstable pelvic ring fractures with little risk of morbidity. PMID- 11431628 TI - [Metaphyseal and diaphyseal modular femoral stems implanted without cement]. AB - PURPOSE OF THE STUDY: We assessed outcome after total hip arthroplasty (THA) using a dual metaphyseal-diaphyseal modular femoral stem with hydroxyapatite coating on the metaphyseal portion only. Implanted without cement, this stem was adaptable to all the anatomic morphotypes defined by the Noble canal flare index. MATERIAL AND METHODS: THA was performed in 116 patients (124 hips), mainly for degenerative joint disease (80% for dysplasia). Mean age was 61.2 years. Follow up was 6.9 years (72-108 months). RESULTS: There were no preoperative complications excepting 3 cases of neck fissuration without clinical consequence. There was no trochanteric fracture. We had two early and one late dislocations. The Postel Merle d'Aubigne score improved from 8.09 to 17.27. Clinical outcome was not influenced by patient age, weight or morphotype. Radiologically, signs of bone ingrowth were found in more than 80% of the cases. Lucent lines were seen in only 3 cases. There was a single case of migration. No revision was needed among the cases with ossifications (23%, 22% Brooker I) and no femur revisions were required. There was no mechanical problem involving the metaphyseal-epiphysial junction. DISCUSSION: The dual metaphyseal-diaphyseal modular stem was found to be a safe and effective implant adaptable to all anatomic variations of the femur and providing good primary stability. In our series, 58% of the femurs were "standard" but for one-third of the femurs, the modular stem enabled implantation without cement, particularly in young adults with a dysplasic or funnel-shaped femur. CONCLUSION: The dual metaphyseal-diaphyseal modular stem was found to be most useful for optimizing total hip arthroplasty without cement. PMID- 11431629 TI - [Quantitative 3D anatomy of the lumbar spine]. AB - PURPOSE OF THE STUDY: There are many anatomic descriptions of the spine, but most concentrate on qualitative information. Quantitative data is however important to achieve a better clinical approach, to adapt implant size and to construct geometric models of spine mechanics. MATERIAL AND METHODS: We examined 32 dry spines (160 lumbar vertebrae) obtained from the Orfila Museum anatomy laboratory at the Saints-Peres School of Medicine in Paris. We drew 178 landmarks on the surface of each vertebra and recorded the 3D spatial coordinates of each point using a Fastrack electromagnetic system operating with +/- 0.2 mm precision. The coordinates of the digitalized points were expressed in a local x-y-z axis field (x=posteroanterior axis, y=right-left axis, z=caudocranial axis). The origin O was half way between the "centers" of the vertebral plates. After calculating 112 linear, angular and surface parameters, results were analyzed with the Statview statistics system. RESULTS: All parameters exhibited gaussian distribution. The transpedicular vertebral depth, corresponding to the maximal penetration of a pedicular screw before touching the anterior wall, was nearly constant: 48 mm (mean). The mean height of the pedicle was approximately 16 mm for L1 to L4 and 21 mm for L5. Pedicle width was 7 mm for L1 and L2 then rapidly widened to reach 10 mm for L5. It was noteworthy that the narrowest pedicle (4 mm) was found in 10% of the L1 vertebrae. There was an exponential rise in the sagittal tilt of the pedicles from L1 to L5, measuring approximately 8 degrees for L1 and rising to 24 degrees for L5. DISCUSSION: Our sample of human spines with unknown clinical characteristics (age, sex) is representative of anatomy laboratory populations, generally composed of subjects over 40 years of age, and is thus adapted for studies of the degenerative spine. Our findings are generally in agreement with data reported in the literature and also provided complementary quantitative data concerning the transpedicular vertebral depth that was found to be a rather constant feature of the lumbar spine. It measured between 40 and 56 mm for 95% of the study population. The dimensions of the pedicle is particularly important: the width must be known to determine the size of pedicle screws; it measured between 7 and 12 mm in 95% of the population. The largest mean cross section of the pedicle was found for L5 (82mm(2)), but measured less than 60 mm(2) in 10% of the vertebrae, suggesting predisposition to spondylolysis. PMID- 11431630 TI - [Knee arthroscopy in children]. AB - PURPOSE OF THE STUDY: Between 1990 and 1998, 110 knee arthroscopies were performed in children. We analyzed the epidemiology and diagnostic data and studied the correlation between clinical and radiographic findings and the final diagnosis after arthroscopy in order to establish a consensus on use of knee arthroscopy in children. MATERIAL AND METHODS: We made a retrospective analysis of 110 knee arthroscopies performed in children, classing the patients in three age groups: 0-5 years, 5-10 years, 10-17 years. Clinical and radiological findings were compared with the arthroscopy findings. RESULTS: One or more arthroscopies were performed in 56 boys and 48 girls. Mean age at the time of the procedure was 12 years 4 months. There were 11 children aged 0-5 years, 14 aged 5 10 years and 85 aged 10-17 years. The main pathology was arthritis in the 0-5 year and 5-10 year age groups. Trauma was more frequent in the older children. Knee arthroscopy was found to be normal in 19 cases. DISCUSSION: For most surgery teams, arthroscopy is indicated for arthritis of the knee. Arthroscopy may also be needed for hemarthrosis. In these contexts, arthroscopy is both a diagnostic and therapeutic procedure. Our analysis demonstrates that emergency arthroscopy is only warranted for free floating osteochondral fractures and fractures of the tibial articular surfaces, with the exception of the tibial spines. Arthroscopy may be performed later in other cases after careful physical examination and radiographic series. We had 19 normal arthroscopies and 10 that showed femoropatellar chondropathies and plicas that could explain knee pain. We recommend arthrography before arthroscopy to avoid unnecessary procedures. CONCLUSION: Arthritis of the knee is an excellent indication for arthroscopy. Painful and acute hemarthrosis requires attentive physical exams and x-rays before making the decision for surgery. PMID- 11431631 TI - [Supracondylar femoral extension osteotomy for knee flexion contracture correction in poliomyelitic conditions]. AB - PURPOSE OF THE STUDY: Knee flexion contracture due to quadriceps paralysis is a major handicap in poliomyelitis patients. The patient has to stabilize the knee with the ipsilateral hand to achieve weight bearing and the deformed knee precludes use of orthopedic devices. Extension can be achieved with supracondylar femoral osteotomy if the knee flexion contracture is less than 30 degrees. We assessed functional and anatomic outcome. MATERIAL AND METHODS: We reviewed the files of 87 patients who had undergone 93 supracondylar femoral osteotomies for knee flexion contracture (6 bilateral cases); mean age was 18 years and mean flexion was 25 degrees. The surgical correction was achieved by diaphyseal metaphyseal impaction with resection of an anterior wedge and preservation of the posterior component of the articulation. If some gluteus maximus activity was retained and the tibiotarsal joint was in a slightly equine position, weight bearing in a stable locked position became an automatic postural event even in case of total paralysis of the quadriceps. Osteotomy was not possible if the contracture flexion was greater than 30 degrees due to excessive tension on the vaculonervous bundles. The procedure was equally impossible in children under 12 years of age due to the risk of recurrence subsequent to migration and callus remodeling with bone growth. RESULTS: Complete extension of the knee was achieved peroperatively in all cases. The most serious complications were three cases of septic arthritis that led to an irreducible stiff knee. In addition, we had two cases of transient paralysis of the common fibular nerve that recovered spontaneously. Bone fusion was achieved in all cases within 30 days. Recurrent flexion contracture was observed in 5 cases and required a revision using the same procedure in 3 or them. Postoperatively, the amplitude gained in knee extension corresponded to the amplitude lost for flexion. Sixty-three patients were able to walk independently without manual stabilization and a knee extension orthesis could be installed for 19 others. Three patients were still unable to walk despite the correction of the knee flexion contracture due to failure of poorly accepted orthopedic devices. DISCUSSION: Several conservative methods (physiotherapy, manipulations, successive corrective casts) and surgical procedures (release of posterior soft tissues, Ilizarov technique) have been proposed for the correction of paralytic knee flexion contracture. Supracondylar femoral osteotomy for extension can be useful after the end of growth if the flexion contracture remains below 30 degrees. The procedure is simple and morbidity is relatively low compared with the regularly successful results. When the flexion contracture exceeds 30 degrees, the supracondylar osteotomy cannot be employed due to the risk of stretching the vasculonervous bundles and due to the instability and disorganization of the lower femur. Progressive correction can be proposed for these patients: regular monitoring of the neurological and vascular situation is required. Functional improvement is considerable after correction of knee flexion contracture. The patients can walk more easily, no longer need to stabilize their knee with their hand, and can benefit from orthopedic devises due to the more favorable biomechanical conditions. PMID- 11431632 TI - [Single-compartment knee arthroplasty prosthesis and idiopathic necrosis of the medial tibia surface]. AB - PURPOSE OF THE STUDY: Idiopathic necrosis of the medial articular surface of the tibia is exceptional. Diagnosis is quite difficult and often made late. Among the different treatments proposed, we preferred single-compartment arthroplasty. MATERIAL AND METHODS: We report 8 cases in women with a mean age of 71.1 years. Diagnosis was suspected due to drug-resistant knee pain, particularly frequent at night, initially with radiographically normal knees. The first radiographic signs, seen 3 months after the onset of pain, were pathognomonic for osteonecrosis evidencing subchondral defects of the tibial surface with a dense peripheral rim and apparently "sequestered" in a notch. Bone scintigraphy evidenced intense uptake in the medial compartment. MRI confirmed the diagnosis evidencing a band of low intensity signals completely surrounding a sequestered zone reaching the cortical. This band was stable and irreversible. In 5 cases CT scan and in 3 cases tomography identified the width and height of the necrotic area that was limited to the medial compartment in all cases. All patients were treated with a single compartment implant. The diagnosis of necrosis was confirmed at pathology. RESULTS: At 4,6 years of mean follow up all patients had an excellent outcome, "forgetting" their knee. No lucent lines developed along the femoral or tibial implants. DISCUSSION: Necrosis of the medial articular surface of the tibia is exceptional and often diagnosed late by bone scintigraphy or MRI. Surgical treatment is usually based on tibial osteotomy for valgisation or a single or three-compartment prosthesis. In our 8 cases, the necrosis was limited to the medial compartment, warranting our therapeutic option. PMID- 11431633 TI - [The origin of femoral trochlear dysplasia: comparative anatomy, evolution, and growth of the patellofemoral joint]. AB - We performed a comparative analysis of the femoropatellar morphology examining the evolutionary aspects to search for the origin of trochelar dysplasia. Trochlear dysplasia is frequent in the human population and, when associated with morphological and positional abnormalities of the patella, can lead to patellar pain syndrome in minor cases or patellar dislocation in severe cases. There is no strict relationship between the observed anomalies and clinical expression. The shape of the articular surfaces is variable in mammals depending on their type of locomotion: unguligrade, digitigrade, plantigrade. In greater apes, the femoral diaphysis is straight and the trochlea is flat and symmetrical. The patella does not tend to dislocate laterally since the knee under load is always flexed. In human adults, the femoral diaphysis has a valgus obliquity angle of 8 degrees to 10 degrees. Consequently, the trochlea has a deepened sulcus and an elevated lateral lip, avoiding lateral patellar dislocation, especially during initial knee flexion. In the human newborn, the femoral diaphysis is vertical. As the child starts walking, the femoral obliquity angle develops between 1 and 7 years of age, inducing a secondary valgus of the extensor apparatus. This obliquity does not develop in non-walking children. Fossil femurs of australopithecus demonstrate that a high obliquity angle had appeared more than 3 million years ago, but also exhibit a poorly deepened trochlea and a slight elevation of the lateral facet. At 1.8 million years, the fossils have the oblique diaphysis, the strongly deepened sulcus and the strongly elevated lateral facet. The obliquity angle of the femoral diaphysis is the leading feature which initiated the later modifications of the patellofemoral joint that over 3 million years were never inscribed in the human genoma. Lateral trochleal lip and deep sulcus are features that were first acquired, then once selected, genetically assimilated, and now appear on the fetal cartilaginous epiphysis. PMID- 11431634 TI - [Dislocation of the knee with isolated posterior cruciate ligament rupture]. AB - We report a case of posterior femorotibial dislocation with an isolated lesion of the posterior cruciate ligament. Outcome was favorable after reduction and conservative treatment for six weeks and was satisfactory at six months follow up. Acceptable functional results can be obtained after non-operative treatment of this very severe knee injury. PMID- 11431635 TI - [Bifocal avulsion of the patellar tendon in an adult: a case report]. AB - PURPOSE OF THE STUDY: Bifocal proximal and distal avulsion of the patellar tendon is a rare event in adults with, to our knowledge, no previous report in the literature. Exceptional bifocal avulsion of the patellar tendon is generally related to an underlying bone or systemic disease. We report the case of a 90 year-old diabetic patient who developed simultaneous avulsions of the patellar and tibial insertions. CASE REPORT: A 90-year-old woman with noninsulin-dependent diabetes mellitus fell on her left knee and was subsequently unable to extend the knee despite normal quadriceps contraction. Radiographically, the knee joint showed a transverse fracture of the lower third of the patella and a fracture of the anterior tibial tuberosity, evidencing rupture of the quadriceps tendon. Signs of osteoarthritis were minimal. The patient underwent surgery for osteosynthesis of the patella with two vertical pins and a figure-eight wire cerclage in addition to fixation of the anterior tuberosity of the tibia with 3 staples and wire cerclage. The patient recovered well and was walking six months later. DISCUSSION: Bifocal disruption of the extensor system is a rare event in adults. It is usually associated with an underlying bone disease (Paget's disease, osteomalacia, osteitis deformans) or systemic disease (lupus erythematosus, rheumatoid arthritis, hyperparathyroidism, Wilson's disease), but bilateral spontaneous disruption may occur. There have been two reports of patellar full-thickness tendon tears in diabetic patients but no case of a bipolar avulsion involving both insertions. Concurrent forces applied to the ligament were probably the cause of this unique type of patellar tendon avulsion. CONCLUSION: This is the first report of a bifocal disruption of the patellar tendon. Different underlying diseases may be involved. Treatment consists in double fixation. PMID- 11431636 TI - [Vertebral hydatidosis: three case reports]. AB - PURPOSE OF THE STUDY: Vertebral hydatidosis, the most frequent localization of skeletal hydatidosis, has an ominous prognosis with the risk of progressive but permanent neurological damage. There is no consensus on appropriate management. Treatment is often unsuccessful with frequent recurrence. The purpose of this study was to assess our experience with this rare disease to identify diagnostic, therapeutic and prognostic features. MATERIAL AND METHODS: We reviewed the charts of three patients with vertebral hydatidosis treated at the department of orthopedics at the Casablanca children's hospital between January 1989 and January 1998. Diagnosis was made early in the first patients, allowing complete excision of the hydatid cyst. Follow-up was uneventful 10 years after surgery. The second patient presented after a long disease course and had definitive neurological complications despite treatment. The lesions were extensive in the third patient and cure could not be achieved although the patient remained asymptomatic. DISCUSSION: Vertebral hydatidosis is a severe disease causing frequent neurological complications. Surgical treatment, though difficult, is the only efficient option. Complete recovery can be achieved after surgical excision of lesions diagnosed early. Prevention is the best therapeutic strategy in endemic areas. PMID- 11431637 TI - [Echinococcosis of the pelvic bone: four cases]. AB - We report four cases of echinococcosis of the pelvic bone in 3 women and a man aged 30 to 55 years. Disease spread was wide, involving the entire hemipelvis in one case and the sacrum in another making surgical excision difficult. In two cases, computed tomography was highly contributive to diagnosis and assessment of local extension. We used surgical excision and hydrogen peroxide sterilization for the first two case. For the two last cases, we used albendazole in four 4 week cures at a 2-week interval before and after surgery. The clinical course was favorable to these two cases at 3 and 4 years. The albendazole-surgery combination appears to be the best therapeutic option. PMID- 11431638 TI - [Ovarian function after total simple hysterectomy]. AB - BACKGROUND: The aim of this prospective study is to evaluate the ovarian function and its duration after total simple hysterectomy. METHODS: This research was carried out on thirty regularly menstruating patients who underwent simple hysterectomy. Ovarian function has been evaluated by measuring FSH, E2, PP at 6 and 24 months from hysterectomy or from the first observation at the beginning of the study. RESULTS: None of the patients showed ovarian failure at 6 months. Nine patients (average age: 48,8 years) showed ovarian failure at 24 months. CONCLUSIONS: The conclusion is drawn that the cessation of ovarian activity is not influenced by hysterectomy: it rather depends on the age of the patient. PMID- 11431639 TI - [Prevention of gestational hypertension with calcium, linoleic acid, mono and polyunsaturated fatty acid supplements]. AB - BACKGROUND: The aim of this study was to evaluate the benefits of calcium, linoleic acid and mono- and polyunsaturated fatty acids in the prevention of gestational hypertension. METHODS: Between January 1999 and June 2000, 65 primagravidas with a risk of gestational hypertension between the 20th and 24th week of pregnancy were recruited using Doppler velocimetry of the uterine arteries and Holter 24-h pressure monitoring. Patients with diabetes, hypertensive gestosis and previous renal pathologies were excluded. The patients were divided into 2 unequal groups: Group A (40 patients) was treated with calcium, linoleic acid, and mono and polyunsaturated fatty acid supplements; Group B (25 patients) received no treatment. The patients in both groups underwent Doppler velocimetry of the uterine arteries and Holter pressure monitoring at 28-30-32-35 weeks of gestation. RESULTS: Calcium linoleic acid and mono and polyunsaturated fatty acid supplements were effective in significantly reducing the incidence of gestational hypertension (p>0.05); in fact 90% of patients in Group A underwent a physiological evolution of pregnancy compared to only 10% affected by gestational hypertension. In the control group the incidence of this pathology was much higher (32%). CONCLUSIONS: The authors conclude that calcium linoleic acid and mono and polyunsaturated fatty acid supplements are useful and effective in the prevention of gestational hypertension in women at risk for this pathology. PMID- 11431640 TI - [Effects of metformin on insulin resistance and on ovarian steroidogenesis in women with polycystic ovary syndrome]. AB - BACKGROUND: Polycystic ovary syndrome is considered the most frequent endocrine abnormality during the fertile age. Hyperinsuli-nemia, typical of this syndrome, alters the functionality of the cytochrome P450 c 17 alfa and this causes an exaggerated secretion of male hormones produced by the female gonad. Objective of this study was to verify if the reduction of the circulating insulin levels, obtained through therapy with metformin, caused the reduction of LH levels, LH:FSH ratio, of testosterone and androstenedione levels, but also of cholesterolemia, triglyceridemia, BMI, and naturally of insulinemia, glycemia, as well as an increase in HDLC (high density lipoprotein cholesterol). METHODS: The presence of insulin-resistance and hyperinsulinemia, in 15 women aged between 20 and 30 with BMI >26 kg/m2, has been verified with test loaded with glucose; 500 mg of metformin have been given to these women three times a day before meals for 12 weeks. LH and FSH levels, LH:FSH ratio testosterone, androstenedione, progesterone, estradiol levels, cholesterolemia, trygliceridemia, HDLC and BMI have been considered in each patient before and after therapy. RESULTS: A statistically significant modification of all measured parameters was observed but the most important data is the reduction of the androgens. CONCLUSIONS: The reduction of insulinemia seems to be actually associated with a reduction of circulating androgens in women with polycystic ovary syndrome. As well as the reproductive function, this insulin-sensitizing agent has the further <> advantage of a possible favourable effect also on the complications of the polycystic ovary syndrome specifically connected with: glucose-intolerance, dislipidemia, arteriosclerosis and hypertension. To consider the effects on the complications of the syndrome it would be necessary to give the drug for a period longer than 12 weeks. PMID- 11431641 TI - [Prenatal diagnosis of beta-thalassemia. A study based in Calabria]. AB - BACKGROUND: The aim of this study was to underline, given the well-known incidence of beta thalassemia in Calabria, the possibility of establishing a prevention programme based on an increased awareness among the population using information and health education, genetic consultancy to identify high-risk subjects and prenatal diagnosis. METHODS: Between January 1992 and December 1999, we analysed 181 high-risk couples for beta thalassemia using chorionic villi sampling (CVS) performed with an echoguided transcervical or transabdominal route. A steady rise was observed over the years in the number of couples asking for prenatal diagnosis, thus demonstrating that patients and doctors are increasingly aware of the importance of a correct prenatal diagnosis. The variability of molecular defects found and the number of complications linked to the technique used are underlined. RESULTS: Prenatal diagnosis revealed 46 fetuses with the disease (24.8%), 97 heterozygotes (53.2%) and 41 healthy fetuses (22%). Only one malformation was observed in the 41 healthy fetuses. The authors also report the incidence of complications linked to CVS. In overall terms, 2.2% of pregnancies ended in spontaneous abortions (4 cases) and 2.75% with preterm births (5 cases). These complications could not be correlated with gestational age at the time of biopsy or the number of attempts made to obtain an appropriate sample. PMID- 11431642 TI - [HRT: state of the art]. AB - Hormone replacement therapy (HRT) in postmenopause represents the most advanced frontier of preventive medicine in a rapidly evolving society that aims to emphasise, today as in the past, the leading role of women. While modern medicine has extended the average life expectancy of women today, it is now the task to enhance the quality of these extra years. HRT prevents cardiovascular disease, osteoporosis, disorders relating to changes in pelvic connective tissue and genitourinary tissues, and it can also have a positive influence on the psycho affective sphere and, perhaps, alterations in cognitive capacity. Cultural delays and alternate phases of optimism and alarm are not always backed by solid scientific knowledge. Epidemiological research over the past years has underlined the oncological risk of using estrogens, even if associated with progestin, without the necessary methodological clarity and efficacy. From the data reported in the international literature and on the basis of over ten years experience accumulated at Department B of Gynecology and Obstetrics of Turin University, it appears that the oncogenic risk in women taking HRT is not significantly higher if the indications are strictly respected and, when necessary, progestin is associated with estrogen. PMID- 11431643 TI - [Hemolytic uremic syndrome in twin pregnancy at 32 weeks gestation with HELLP syndrome. Case report]. AB - Renal failure occurring in pregnancy or post partum is an unusual but well described complication. Acute renal failure seems to be associated more often with HELLP syndrome rather than with pre-eclampsia or chronic hypertension. Probable overlapping of HELLP and hemolytic uremic syndrome in pregnancy or postpartum should be taken into consideration when treating pregnant women who show signs of proteinuria, hypertension, hematuria, increase of reticulocytes, decrease of haptoglobin with thrombocytopenia and microangiopathic hemolytic anemia. Our case refers to a 32 year old woman at 32 weeks gestation in twin pregnancy who presented with HELLP syndrome and renal failure. Immediately postpartum oliguria was noted and the laboratory analyses suggested the coexistence of HELLP and hemolytic uremic syndrome. In patients with gestosis and/or HELLP syndrome presenting oliguria combined with a decrease of hemoglobin level not due to intraoperative hematic leaks it is always necessary to ask for haptoglobin dosage. In treating hemolytic uremic syndrome it is very important to use a high dosage of plasma and sometimes plasmapheresis. HELLP syndrome contributes to various complications which are sometimes responsible for kidney or maternal mortality. In treating these patients early diagnosis combined with a specific treatment can considerably reduce kidney and maternal mortality. PMID- 11431644 TI - [Endometriosis on laparotomy scar after cesarean section]. AB - Cutaneous endometriosis is an infrequent pathology whose etiopathogenesis is still uncertain. A case of cutaneous endometriosis at the surgical scar level following cesarean section is reported. Moreover, the aspects which might facilitate the diagnostic approach and a correct therapeutic management are underlined. PMID- 11431645 TI - [A case of atypical bicornate unicollis uterus treated with metroplasty according to Jones]. AB - The aim of the study is to report on the result achieved on uterus bicornate unicollis that after hysterosalpingography proved atypical in the isthmic section of the left horn because it was filiform and had irregular walls. The metroplasty technique was performed, applying the Jones technique, on a patient affected by infertility for six years, with uterine malformation. Hysterosalpingography evidenced an atypical malformation: a unicollis-bicornis uterus that was atypical in the isthmic section of the left horn, treated with the technique of Jones because it was filiform. Diagnostic laparoscopy visualized two symmetrical and separated horns. Metroplasty was performed applying Jones technique, by removing the isthmic miometrium of the left horn in order to widen the communication with the upper part. The result of postoperative hysterosalpingography, performed 10 months after intervention, showed a single uterine cavity with irregular walls that allowed the evolution of a pregnancy that was prematurely concluded after thirty-two weeks on the birth of a live, vital fetus. Infertility is probably associated with the complexity of uterine malformation. We can, therefore, underline that laparotomic metroplasty is an effective technique that can be performed in particular cases. PMID- 11431646 TI - [Extraumbilical insertion of the laparoscope in abdominal adhesion]. AB - Palmer s point must be considered a safe alternative site to insert the Verres needle and the primary trocar. It s possible, also, to perform adhesiolysis by this way before to insert the bigger trocar in umbilical site. Our experience shows that this technique can avoid intestinal, omental, injury if the trocar was inserted directly using the umbilical site: that in patients with previous surgery. PMID- 11431647 TI - [Luteal phase support in assisted reproductive cycles using either vaginal (Crinone 8) or intramuscular (Prontogest) progesterone: results of a prospective randomized study]. AB - BACKGROUND: To compare local tolerance and patients compliance to intravaginal and intramuscular progesterone administration. METHODS: Ninety-nine patients have been randomised to receive either intravaginal Crinone, 90 mg/day (n=51) or intramuscular Prontogest 50 mg/day (n=48) for luteal supplementation in IVF/ICSI cycles. Local and systemic side effects as well as pattern of menstrual bleeding were reported on a self administered questionnaire. Progesterone levels were evaluated pre-treatment, in the mid-luteal phase and the day of pregnancy test. RESULTS: Patients age, BMI, duration and causes of infertility were comparable in the two treatment groups. All parameters of ovarian response as well as pregnancy rates did not show significative difference in the two groups. A significative larger number of patients assigned to intravaginal support were free from side effects. Furthermore side effects, when reported, resulted significantly more severe in the intramuscular group. In the non pregnant patients menstrual flow appeared significantly earlier in those treated with vaginal progesterone (p<0.001). CONCLUSIONS: Crinone 8 is a good alternative to parental progesterone for luteal support in ART cycles. It is well tolerated but it is linked to an earlier appearance of menstrual flow in non conceptional cycles. PMID- 11431648 TI - Interpreting the exercise electrocardiogram in patients either on beta-blocking drugs or with chronotropic incompetence. PMID- 11431649 TI - The emerging role of myoglobin for risk stratification. PMID- 11431650 TI - Antithrombotic therapy after prosthetic cardiac valve implantation: potential novel antithrombotic therapies. PMID- 11431651 TI - A tale of two countries: Insights from the differences in Canadian/American patterns of care for patients with acute coronary syndromes. PMID- 11431652 TI - Statistical issues relating to international differences in clinical trials. PMID- 11431653 TI - Myoglobin levels at 12 hours identify patients at low risk for 30-day mortality after thrombolysis in acute myocardial infarction: a Thrombolysis in Myocardial Infarction 10B substudy. AB - OBJECTIVE: We sought to identify, by use of serum cardiac markers, patients at low risk for 30-day mortality after ST-segment elevation myocardial infarction. BACKGROUND: Baseline cardiac markers are currently used to identify patients at increased risk for short-term events. We hypothesized that serum markers measured after treatment could identify patients at low risk for 30-day mortality. METHODS: A total of 839 patients from the Thrombolysis in Myocardial Infarction (TIMI) 10B study had myoglobin, cardiac-specific troponin-I, creatine kinase (CK) MB measurements at the following time points; baseline, 90 minutes, and 3 and 12 hours after thrombolysis. By use of receiver operating characteristic analysis, thresholds were derived to predict 30-day mortality with at least 95% negative predictive value. RESULTS: Ninety minutes after thrombolysis myoglobin was superior to troponin-I or CK-MB in identifying patients at low risk for mortality. The 30-day mortality for 12-hour myoglobin < or = 239 ng/mL was 1.4% compared with 9.1% for levels > 239 ng/mL (P < .001). For 12-hour troponin-I (threshold 81.5 ng/mL), mortality was 1.9% versus 6.6% (P = .001) if above threshold; similarly for CK-MB at 12 hours (threshold 191 ng/mL) it was 3.3% versus 7.9% (P = .02). Multivariate analysis of baseline and posttreatment cardiac markers, age, sex, infarct artery location, and 90-minute TIMI flow grade identified only 12-hour myoglobin among the cardiac markers as independently predicting a low 30-day mortality (odds ratio 0.11, 95% confidence interval 0.02 0.50, P < .004). CONCLUSION: Serum cardiac markers can identify greater than two thirds of patients at low risk for 30-day mortality. A low 12-hour myoglobin level (< or = 239 ng/mL in this substudy) identifies such patients at low risk and could potentially assist in early risk stratification and triage after ST segment elevation myocardial infarction. PMID- 11431654 TI - Post-myocardial infarction risk stratification in elderly patients. AB - BACKGROUND: The purpose of this study was to examine the use of post-myocardial infarction (MI) risk stratification in the elderly. Although expert panels have recommended risk stratification after MI, limited data are available on whether patients actually undergo suggested testing. In particular, concern has been raised that the elderly, who are at high risk for recurrent ischemia and short term death, are not referred as often as younger patients for post-MI testing. METHODS: We studied the records of 192,311 Medicare patients (age > or = 65 years) admitted with MI between January 1992 and November 1992. By combining Medicare part A and part B data, we created a longitudinal record of patient care within 60 days of an MI admission. We describe the pattern of post-MI testing for ischemia and left ventricular function and outcomes as a function of patient age. RESULTS: Patients > or = 75 years of age were significantly less likely than patients 65 to 74 years of age to have either cardiac catheterization (17% vs 43%) or any test for coronary artery disease severity (24% vs 53%). They were also less likely to have a test of left ventricular function (61% vs 76%). Even after adjustment for baseline characteristics, older patients remained less likely than younger patients to have an assessment of coronary artery disease severity (odds ratio, 0.44) or left ventricular function (odds ratio, 0.65). CONCLUSIONS: Post-MI risk stratification declines with age and falls short of recommendations in our nation's elderly. This lack of testing may result in lost opportunities for therapeutic interventions in this high-risk group. PMID- 11431655 TI - Predictors of left ventricular function after acute myocardial infarction: effects of time to treatment, patency, and body mass index: the GUSTO-I angiographic experience. AB - BACKGROUND: Despite the significant survival benefit associated with successful reperfusion therapy for acute myocardial infarction, global indices of outcome left ventricular function, such as ejection fraction, have often demonstrated little or no improvement. Although these measurements are confounded by numerous clinical, physiologic, and angiographic variables, no comprehensive analysis of this issue in a large series of patients is available. We used the Global Utilization of Streptokinase and Tissue Plasminogen Activator for Occluded Coronary Arteries (GUSTO-I) database to better understand this phenomenon by determining independent predictors of left ventricular function and their interplay with regard to outcome ventricular function and improvement in function during the initial postinfarction week. METHODS: Ninety-minute and 5- to 7-day posttreatment global and regional indices derived from left ventriculograms were analyzed from a population of 676 patients. These observations were combined with clinical data to describe independent determinants of ventricular function outcome. RESULTS: Clinical factors predictive of global and regional ventricular function as well as improvement in function between 90 minutes and 5 to 7 days included time to treatment, early infarct-related artery flow grade, and body mass index. These same factors contribute significantly to compensatory hyperkinesis of the noninfarct zone, which is critical to maintenance of global ventricular function during this time period. CONCLUSIONS: The ventricular function benefits of early complete reperfusion after myocardial infarction are readily demonstrable after adjustment for multiple covariables and include (1) maintenance of global ventricular function and (2) prevention or delay in ventricular dilatation. PMID- 11431656 TI - ST-segment depression in lead aVR predicts predischarge left ventricular dysfunction in patients with reperfused anterior acute myocardial infarction with anterolateral ST-segment elevation. AB - BACKGROUND: Patients with an anterolateral acute myocardial infarction (AMI) have a worse prognosis, and those with additional inferolateral wall involvement might be higher risk because of more extensive area at risk. Lead -aVR obtained by inversion of images in lead aVR has been reported to provide useful information for inferolateral lesion. METHODS: We examined the relation between ST-segment deviation in lead aVR on admission electrocardiogram (ECG) and left ventricular function in 105 patients with an anterolateral AMI undergoing successful reperfusion < or = 6 hours after onset. Patients were classified according to ST segment deviation in lead aVR on admission ECG: group A, 23 patients with ST elevation of > or = 0.5 mm; group B, 47 patients without ST deviation; and group C, 35 patients with ST depression of > or = 0.5 mm. RESULTS: There were no differences among the 3 groups in age, sex, or site of the culprit lesion. In groups A, B, and C, the peak creatine kinase level was 3661 +/- 1428, 4440 +/- 1889, and 6959 +/- 2712 mU/mL, and the left ventricular ejection fraction (LVEF) measured by predischarge left ventriculography was 54% +/- 9%, 48% +/- 7%, and 37% +/- 9%, respectively(P < .01). During hospitalization, congestive heart failure occurred more frequently in group C than in groups A or B (P < .05). ST segment depression in lead aVR had a higher predictive accuracy than other ECG findings in identifying patients with predischarge LVEF < or = 35%. CONCLUSIONS: We conclude that in patients with an anterolateral AMI, ST-segment depression in lead aVR on admission ECG is useful for predicting larger infarct and left ventricular dysfunction despite successful reperfusion. PMID- 11431657 TI - Continuous ST monitoring: a bedside instrument? A report from the Assessment of the Safety of a New Thrombolytic (ASSENT 2) ST monitoring substudy. AB - BACKGROUND: Continuous ST monitoring has during the past decade become widespread in coronary care units (CCU) and is now even recommended by international task forces to survey patients with acute coronary syndromes. ST monitoring has also been shown to be a well-validated technology to predict prognosis and coronary artery patency in patients with acute ST-elevation infarction. However, all evaluations in previously presented trials have been made inside a core laboratory by specially trained personnel. METHODS: In the Assessment of the Safety and Efficacy of a New Thrombolytic (ASSENT 2) ST-monitoring substudy, 448 patients with ST-elevation infarction from 22 hospitals were monitored by vectorcardiography. The local nurse responsible for the patient made an evaluation of the ST trend curve. This was compared with a blinded core laboratory evaluation and examined versus 30-day mortality. RESULTS: No significant differences were found between the local and central evaluations of ST-segment recovery. The accuracy of the local evaluation, compared with the central one, in deciding whether patients had > or = 50% ST-segment recovery at 60 or 90 minutes was > 90%. A large low-risk group (30% of the patients) with 0.8% 30-day mortality could be identified by the local evaluation. CONCLUSIONS: ST monitoring with vectorcardiography can accurately be done in the clinical setting. The local evaluation was as least as accurate as the core laboratory evaluation in predicting prognosis. PMID- 11431658 TI - Experimental pain and psychologic status of patients with chest pain with normal coronary arteries or ischemic heart disease. AB - BACKGROUND: The cause of chest pain in patients with a normal coronary angiogram (NCA) remains an enigma. Also, it is unclear whether psychosocial factors play a role in the etiology of chest pain in these patients. The objective of the current study was to compare psychosocial factors, clinical pain, and responses to experimental pain in NCA patients, patients with ischemic heart disease (IHD), and healthy control subjects. METHODS: Pain intensity, threshold, and tolerance to cold pressor pain were assessed in 30 NCA patients, 30 IHD patients, and 30 healthy control subjects matched for age, sex, and sociodemographic factors. All subjects completed questionnaires measuring a number of psychosocial factors, including stress, anxiety, depression, extroversion, and neuroticism. NCA and IHD patients also completed questionnaires assessing clinical pain responses and pain coping strategies. RESULTS: With the exception of a lower tolerance to cold pressor pain of IHD patients (P <.05), no significant differences were found between NCA and IHD patients with respect to other clinical pain measures, psychosocial measures, pain-coping strategies, and other pain-related behaviors. Healthy control subjects differed significantly (P <.05) from both IHD and NCA patients with respect to maximum cold pressor pain, depression, and state anxiety and from IHD patients with respect to intensity of cold pressor pain, threshold to cold pressor pain, and perceived stress. CONCLUSIONS: The results suggest that higher scores on various psychosocial measures in both chest pain groups are related to their pain, rather than being the cause of pain, and do not support a psychogenic explanation for chest pain in the presence of normal coronary arteries. PMID- 11431659 TI - Clinical outcome of a cohort of patients eligible for therapeutic angiogenesis or transmyocardial revascularization. AB - BACKGROUND: Newer methods of coronary revascularization are being investigated in patients who are not candidates for coronary artery bypass grafting or percutaneous intervention. Our objective was to determine the proportion of patients eligible for newer methods of revascularization and determine their 1 year clinical outcome. METHODS: Five hundred consecutive charts and angiograms from patients undergoing diagnostic angiography for coronary artery disease from January to May of 1998 were reviewed to assess the suitability for revascularization. Patients ineligible for conventional revascularization were followed up for 1 year. RESULTS: Fifty-nine patients of the 500 studied were identified who had refractory ischemia but were not candidates for traditional revascularization. The 59 patients ineligible for traditional methods of revascularization had a rehospitalization rate of 128% (76 total hospitalizations), a 25.5% rate of myocardial infarction (15 of 59), and a mortality rate of 16.9% (10 of 59). CONCLUSIONS: The prognosis of many patients eligible for newer methods of revascularization on maximal medical therapy is poor. PMID- 11431660 TI - The risk of stroke and death in patients with aortic and mitral valve endocarditis. AB - BACKGROUND: Previous studies have generated inconsistent results when attempting to define predictors of stroke and death in patients with endocarditis. We sought to examine the relationship between vegetation 2-dimensional size and stroke in those with infective endocarditis (IE) and to identify differences between aortic valve (AV) and mitral valve (MV) IE with regard to clinical characteristics, echocardiographic findings, stroke, and death. METHODS: We used the Duke Endocarditis Database to examine 145 episodes of definite IE involving the AV, n = 62, or MV, n = 83. A logistic regression model was developed to analyze important variables in predicting stroke, and a Cox proportional hazards model was used in predicting mortality. RESULTS: The mitral valve was infected in 57% of the cases. Vegetations were more commonly detected in patients with MV IE (92.8% vs 66.1%, P =.001) and these MV vegetations were significantly larger (P <.05). Thirty-four of 145 episodes (23.4%) were complicated by stroke. MV IE was associated with a greater stroke rate, 32.5% versus 11.3% (P =.003). Strokes tended to occur early in the course of illness, particularly in MV IE. In the multivariable model, the independent predictors of stroke were MV IE (P =.04) and vegetation length (P =.03). Independent predictors of 1-year mortality were age (P =.02) and vegetation area (P =.048). CONCLUSION: Stroke is more common in patients with MV IE. Vegetation 2-dimensional size and characteristics are important predictors of stroke and mortality. These findings may lead to predictive models that allow physicians to identify high-risk patients who need aggressive treatment strategies to prevent long-term morbidity and mortality. PMID- 11431661 TI - Value of coronary stenotic flow velocity acceleration on the prediction of long term improvement in functional status after angioplasty. AB - BACKGROUND: The coronary flow velocity acceleration at the stenotic site (SVA), defined as a > or = 50% increase in resting stenotic velocity when compared with the reference segment, has been shown to be highly sensitive and specific for the diagnosis of a hemodynamically significant stenosis. In this study, we describe the value of postprocedural SVA for the prediction of a lack of improvement in functional activity at long-term follow-up balloon angioplasty (BA). METHODS: We investigated the improvement in functional activity in patients undergoing single native vessel angioplasty and intracoronary Doppler (before BA, after BA, and again at 6-month follow-up) as part of the Doppler Endpoints Balloon Angioplasty Trial Europe (DEBATE) I trial. Lack of improvement was defined as no change in Duke Activity Status Index (DASI) at 6-month follow-up, whereas SVA was defined as > or = 50% elevation in resting velocity at the treated area compared with the distal measurement. RESULTS: SVA was found more frequently in patients without improvement in DASI (45% vs 31%, P =.03). Similar percent diameter stenosis and coronary flow velocity reserve were observed in patients with and those without improvement in DASI at follow-up. By multivariate regression analysis, the presence of SVA (P = .029; odds ratio, 1.97; 95% confidence interval, 1.07 to 3.63) and an elevated DASI at baseline (P < .001; odds ratio, 1.05; 95% confidence interval, 1.03 to 1.07) were associated with a lack of improvement at follow-up. CONCLUSIONS: The detection of SVA was associated with failure of improvement in functional activity at follow-up after coronary intervention. PMID- 11431662 TI - Incidence and short-term prognosis of late sustained ventricular tachycardia after myocardial infarction: results of the Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto Miocardico (GISSI-3) Data Base. AB - BACKGROUND: There is little epidemiologic information from large multicenter databases on sustained monomorphic ventricular tachycardia occurring after the initial 48 hours of myocardial infarction. METHODS: We reassessed its incidence and short-term prognosis in 16,842 patients with a definite myocardial infarction enrolled in the Gruppo Italiano per lo Studio della Soprovvivenza nell'Infarto Miocardico (GISSI-3) trial. RESULTS: The incidence rate of late sustained ventricular tachycardia by 6 weeks was around 1%. Older age, a history of hypertension, diabetes, and myocardial infarction, nonadministration of lytic therapy, Killip class > I, > or = 6 leads with ST-segment elevation, higher heart rate, and bundle branch block on admission were significantly more frequent among patients with than without late sustained ventricular tachycardia. Patients with ventricular tachycardia had a more complicated course in-hospital and posthospital to 6 weeks than the reference group did. The arrhythmia was associated with a significant excess of pump failure, atrial flutter fibrillation, asystole, atrioventricular block, ventricular fibrillation within the first 48 hours of myocardial infarction, and recurrent ischemic events. Larger left ventricular end-systolic volumes and lower ejection fractions were more frequent among ventricular tachycardia patients than in the reference group by 6 weeks. Death rates by 6 weeks were 35% for patients with ventricular tachycardia and 5% for those without the arrhythmia. Irrespective of the stratification of patients by site and type of infarct and presence/absence of bundle branch block, the occurrence of the arrhythmia was associated with reduced 6-week survival. CONCLUSION: In a proportional hazard regression model late sustained ventricular tachycardia was retained as a strong, independent predictor of 6-week mortality after myocardial infarction (hazard ratio 6.13, 95% confidence interval 4.56-8.25). PMID- 11431663 TI - Efficacy and safety of dofetilide in the prevention of symptomatic episodes of paroxysmal supraventricular tachycardia: a 6-month double-blind comparison with propafenone and placebo. AB - BACKGROUND: Existing drug therapies for paroxysmal supraventricular tachycardia (PSVT) have potentially serious adverse effects. Dofetilide, a pure class III antiarrhythmic agent, may offer an effective and safe alternative for treating PSVT. This study compared the efficacy and safety of dofetilide with that of propafenone and placebo in the prevention of PSVT. METHODS: This multicenter, randomized, placebo-controlled, parallel-group study compared the effectiveness of oral dofetilide 500 microg given twice daily with that of propafenone 150 mg given 3 times a day and placebo in preventing the recurrence of PSVT in 122 symptomatic patients. Episodes of PSVT were documented by symptom diaries and Hertcard (Hertford Medical, Hertfordshire, UK) event recorders. RESULTS: After 6 months of treatment, patients taking dofetilide, propafenone, and placebo had a 50%, 54%, and 6% probability, respectively, of remaining free of episodes of PSVT (P <.001 for both dofetilide and propafenone vs placebo). Both dofetilide and propafenone also decreased the frequency of episodes of PSVT; the median numbers of episodes in the dofetilide- and propafenone-treated groups were 1 and 0.5, respectively, compared with 5 in the placebo-treated group. Dofetilide was well tolerated; no proarrhythmia occurred. Three patients taking propafenone had serious treatment-related adverse effects that required drug discontinuation. CONCLUSIONS: Dofetilide and propafenone were equally effective in preventing the recurrence of or decreasing the frequency of PSVT. PMID- 11431664 TI - Baseline factors predicting early resumption of driving after life-threatening arrhythmias in the Antiarrhythmics Versus Implantable Defibrillators (AVID) Trial. AB - BACKGROUND: In the Antiarrhythmics Versus Implantable Defibrillators (AVID) Trial, patients with ventricular fibrillation or hemodynamically unstable ventricular tachycardia were randomly assigned to receive either an implantable cardioverter-defibrillator (ICD) or antiarrhythmic drug therapy. As part of the trial, patients were asked to participate in a prospective driving survey. The purpose of the survey was to determine what baseline factors and patient characteristics specifically predicted resumption of driving earlier than advised by current guidelines. METHODS: Patients were surveyed anonymously as to their driving habits in the initial period after random assignment and every 6 months thereafter. AVID study coordinators were independently asked to assess their patients' driving status as well. The relation between baseline factors and time to resumption of driving was explored by means of Kaplan-Meier estimates for univariate analyses and the stepwise Cox proportional hazards regression model for multivariate analyses. RESULTS: There were 802 patients who were eligible for assessment of driving status. The majority of patients (58%) resumed driving an automobile within 6 months of their index arrhythmia regardless of whether they received drug therapy or an ICD. By multivariate analysis, patients who were younger than 65 years of age, male, and college educated were more likely to drive early, as were patients whose index arrhythmia was ventricular tachycardia. CONCLUSIONS: Younger, college-educated men and those whose index arrhythmia is ventricular tachycardia are most likely to resume driving <6 months after the initiation of therapy for a potentially life-threatening ventricular arrhythmia. Patients with an ICD did not appear to resume driving later than those who were discharged on antiarrhythmic drugs alone. PMID- 11431665 TI - Impact of prehospital delay on mortality in patients with acute myocardial infarction treated with primary angioplasty and intravenous thrombolysis. AB - BACKGROUND: In patients with acute myocardial infarction treated with thrombolysis, longer times to treatment are associated with increasingly worse clinical outcome. This relation may be different for treatment with primary angioplasty. METHODS: We analyzed the pooled data of the German acute myocardial infarction registries Maximal Individual Therapy in Acute Myocardial Infarction (MITRA) and Myocardial Infarction Registry (MIR) to determine the influence of prehospital delays on hospital mortality rates. Primary angioplasty was performed in 1063 patients and thrombolysis in 7552 patients. RESULTS: In patients treated with thrombolysis, in-hospital time to treatment was constantly 30 minutes median. In patients treated with primary angioplasty, in-hospital time to treatment increased from 60 minutes median up to 87 minutes median with increasing prehospital delay. Hospital mortality rates slightly decreased with increasing prehospital delays in patients treated with primary angioplasty (P for trend =.02). However, in patients treated with thrombolysis, mortality rate was nonsignificantly increased (P for trend =.11). Logistic regression analysis showed no significant difference in mortality rates between primary angioplasty and thrombolysis for prehospital delays of <3 hours. However, when prehospital delay was >3 hours, thrombolysis was independently associated with a higher mortality rate compared with primary angioplasty. CONCLUSIONS: Compared with thrombolysis, primary angioplasty is independently associated with a lower mortality rate in prehospital delays of >3 hours. The reason for this may be a time-dependent loss of efficacy to achieve reperfusion for thrombolysis but not for primary angioplasty. PMID- 11431666 TI - Resource utilization and clinical outcomes of coronary stenting: a comparison of intravascular ultrasound and angiographical guided stent implantation. AB - BACKGROUND: Intravascular ultrasound (IVUS) guided stent implantation studies have demonstrated that inadequate stent implantation can occur despite achieving an optimal angiographic result. Furthermore, IVUS-guided stent implantation has been shown to improve lesional acute gain. However, it is unknown whether the use of IVUS guidance during stent implantation is associated with improved acute and long-term clinical outcomes. Moreover, the additional procedural cost and time incurred with the use of IVUS-directed stent implantation has not been evaluated. Thus the purpose of this study was to determine whether IVUS-guided stent implantation is associated with improved clinical outcomes compared with angiographically guided stent implantation and to evaluate the difference in resource utilization between these respective stent deployment strategies. METHODS: Data were collected on 278 consecutive patients in whom 455 stents were deployed in native coronary arteries. High-pressure (> or = 12 atm) balloon inflations were performed until an optimal angiographic result was obtained. In the angiographically guided group, no IVUS imaging was performed. In the IVUS guided group, IVUS imaging and additional interventions were performed attempting to achieve full apposition, absence of edge tear, and acute gain (lesion lumen area: distal reference lumen area) > or = 0.8 in subsequent IVUS imaging. Total procedure time, fluoroscopy time, contrast media volume, number of balloons, stents, guidewires, guide catheters, and procedural cost were calculated. In hospital abrupt closure rate and 6-month major adverse cardiovascular events (cardiac death, myocardial infarction, target vessel revascularization) rate were obtained. RESULTS: A total of 178 patients underwent IVUS-guided stent placement and 100 patients underwent angiographically guided stent implantation. There was no significant difference in procedure time (107 +/- 49 vs 100 +/- 50 minutes, P = .22), fluoroscopy time (33 +/- 24 vs 30 +/- 18 minutes, P = .36), contrast volume (411 +/- 157 vs 386 +/- 181 mL, P = .23), guide catheters (1.3 +/- 0.8 vs 1.3 +/- 0.6, P = .69), guidewires (1.6 +/- 1.2 vs 1.6 +/- 1.0, P = .99), balloons (2.4 +/- 1.0 vs 2.3 +/- 1.3, P = .58), and stents (1.7 +/- 0.9 vs 1.6 +/- 0.9, P = .42). Intraprocedural cost was significantly higher in the IVUS-guided group, $4142 +/- 1547 verus $3635 +/- 1949 (P = .03), which was primarily related to the cost of the IVUS catheter. However, the in-hospital acute vessel closure rate was significantly lower in the IVUS-guided group, 0.6% versus 4% (P = .04). There was a trend toward lower target vessel revascularization rate in the IVUS-guided group (11% vs 19%, P = .08). By multivariate analysis IVUS use was demonstrated to be an independent negative predictor of cardiac death, myocardial infarction, repeat revascularization, and abrupt stent closure with a relative risk of 0.49 (95% confidence interval of 0.25 to 0.98), and P = .04. CONCLUSIONS: The use of IVUS guidance during stent implantation does not significantly increase procedure time, fluoroscopy exposure, contrast volume, or device utilization. Furthermore, despite the increase in procedural cost, IVUS-guided stent implantation is associated with a significant decrease in the in-hospital abrupt closure rate and a trend toward a lower 6-month target vessel revascularization. PMID- 11431667 TI - Long-term survival in 11,661 patients with multivessel coronary artery disease in the era of stenting: a report from the Alberta Provincial Project for Outcome Assessment in Coronary Heart Disease (APPROACH) Investigators. AB - BACKGROUND: Studies of survival of patients with multivessel coronary artery disease (MVD) in the prestent era suggested that outcomes after coronary artery bypass surgery (CABG) are similar to those after percutaneous coronary intervention (PCI) in subsets of coronary severity. The purpose of this study of the Alberta Provincial Project for Outcome Assessment in Coronary Heart Disease (APPROACH) was to examine the association between treatment and survival up to 5 years in patients with MVD enrolled from 1995 through 1998. METHODS AND RESULTS: Data on patient characteristics were obtained at the time of the initial coronary angiography. Survival was determined through data linkage to the provincial Bureau of Vital Statistics. Risk-adjusted hazard ratios were calculated to compare different treatments. In the 11,661 patients with MVD, CABG was the initial therapy in 3782, PCI in 3540, and medical therapy in 4339. Cumulative 5 year survival was 91.4% with CABG, 91.9% with PCI, and 82.9% with medical therapy (P <.001). Hazard ratios were CABG: medical 0.53 (95% confidence interval [CI] 0.46-0.71), PCI: medical 0.65 (95% CI 0.56-0.74), and CABG: PCI 0.81 (95% CI 0.68 0.96). Analysis across coronary severity groups revealed a benefit of CABG compared with PCI only in the group with severe left main CAD: 0.30 (95% CI 0.17 0.54). CONCLUSIONS: In a multicenter clinical setting, MVD patients treated with revascularization have significantly higher 5-year survival rate than do those treated medically. Risk-adjusted comparison reveals PCI treatment to be associated with long-term survival similar to treatment with CABG in all coronary severity subgroups except the group with severe left main coronary artery disease. Patient selection factors are likely to be contributing to these findings. PMID- 11431668 TI - Diagnostic exercise tests on 4000 consecutive men. AB - OBJECTIVE: Our purpose was to report the prevalence of abnormal treadmill test responses and their association with mortality in a large consecutive series of patients referred for standard diagnostic exercise tests, with testing performed and reported in a standardized fashion. BACKGROUND: Exercise testing is widely performed, but an analysis of responses has not been presented for a large number of consecutive tests performed on patients referred for diagnosis of cardiac disease. METHODS: All patients referred for evaluation at 2 university-affiliated Veterans Affairs Medical Centers who underwent exercise treadmill tests for clinical indications between 1987 and 2000 were determined to be dead or alive according to the Social Security Death Index after a mean 5.9-year follow-up. Patients with established heart disease (ie, prior coronary bypass surgery, myocardial infarction, or congestive heart failure) were excluded from analyses. Clinical and exercise test variables were collected prospectively according to standard definitions; testing and data management were performed in a standardized fashion with a computer-assisted protocol. All-cause mortality was used as the end point for follow-up. Standard survival analysis was performed, including Kaplan-Meier curves and a Cox hazard model. RESULTS: After the exclusions, 3974 men (mean age 57.5 +/- 11 years) had standard diagnostic exercise testing over the study period with a mean of 5.9 (+/-3.7) years of follow-up (64% of all tested). There were no complications of testing in this clinically referred population, 82% of whom were referred for chest pain, risk factors, or signs and symptoms of ischemic heart disease. Five hundred forty-nine (14%) had a history of typical angina. Indications for testing were in accordance with published guidelines. A total of 545 died, yielding an annual mortality rate of 1.8%. The Cox hazard model chose the following variables in rank order as independently associated with time to death: change in rate pressure product, age greater than 65 years, METs less than 5, and electrocardiographic left ventricular hypertrophy. A score based on these variables classified patients into low-, medium-, and high-risk groups. The high-risk group with a score greater than 3 has a hazard ratio of 4 (95% confidence interval 3.82-4.27) and an annual mortality rate of 4%. CONCLUSION: This comprehensive analysis provides rates of various abnormal responses that can be expected in men referred for diagnostic exercise testing at typical Veterans Administration Medical Centers. Four simple variables combined as a score predict all-cause mortality after clinical decisions for therapy are prescribed. PMID- 11431669 TI - Effects of chronotropic incompetence and beta-blocker use on the exercise treadmill test in men. AB - OBJECTIVE: Our purpose was to assess the diagnostic characteristics of the exercise test in patients who fail to reach conventional target heart rates and in patients on beta-blockers. BACKGROUND: Exercise test results are often considered "inadequate" or "nondiagnostic" in patients taking beta-blockers and in patients who do not achieve 85% of their age-predicted maximal heart rate. METHODS: The results of exercise tests and coronary angiography performed to evaluate chest pain in 1282 male patients without a prior history of myocardial infarction, coronary revascularization, diagnostic Q wave on the baseline electrocardiogram, or previous cardiac catheterization were analyzed with respect to beta-blocker exposure and failure to reach 85% age-predicted maximal heart rate. Sensitivity, specificity, and predictive accuracy of exercise testing, as well as area under the curve for the receiver operating characteristic plots were calculated for these subgroups with use of coronary angiography as the reference. The angiographic criterion for significant coronary artery disease was 50% narrowing or greater in one or more major coronary arteries. RESULTS: The population was divided into 4 exclusive groups on the basis of whether they reached their target heart rates and whether they were receiving beta-blockers. Sixty to 40 percent of this clinical population failed to reach target heart rate, of which 24% (n = 303) were receiving beta-blockers and 40% (n = 518) were not. The group of patients who reached target heart rate and were not taking beta blockers was taken as the reference group (n = 409). The group of patients supposedly beta-blocked but who reached the target heart rate (n = 52) had hemodynamic and test characteristics similar to those of the reference group and most likely were not taking their beta-blockers or were not adequately dosed. The prevalence of angiographic coronary disease was significantly higher in the 2 groups failing to reach target heart rate, both in the presence and absence of beta-blockers, compared with the reference group (68% and 64%, respectively, vs 49%, P <.01). Although the areas under the curve of the receiver operating characteristic curves for ST depression of the groups failing to reach target heart rate were not significantly different from the reference group, the predictive accuracy and sensitivity were significantly lower for 1 mm of ST depression in the beta-blocked group who did not reach target heart rate (predictive accuracy of 56% vs 67%, sensitivity of 44% vs 58%, P <.01). The only way to maintain sensitivity with the standard exercise test in the beta-blocker group who failed to reach target heart rate was to use a treadmill score or 0.5 mm ST depression as the criteria for abnormal. CONCLUSION: Sensitivity and predictive accuracy of standard ST criteria for exercise-induced ST depression are significantly decreased in male patients who are taking beta-blockers and do not reach target heart rate. In those who fail to reach target heart rate and are not beta-blocked, sensitivity and predictive accuracy are maintained. PMID- 11431670 TI - Is the dyspnea during adenosine cardiac stress test caused by bronchospasm? AB - BACKGROUND: Adenosine cardiac stress is widely used as an alternative to exercise testing during myocardial perfusion imaging (MPI). Dyspnea often accompanies the adenosine stress test. Although known asthmatic patients are excluded from this test because of the danger of bronchospasm, there is incomplete information regarding the role of factors such as chronic obstructive pulmonary disease (COPD) and tobacco smoking in adenosine-induced dyspnea. PATIENTS AND METHODS: A total of 122 consecutive patients (75 male, 47 female; mean age 59 years, SD 10 years) undergoing pharmacologic stress testing with an intravenous infusion of adenosine also had volume-flow spirometry (Vitalograph Compact, Vitalograph Ltd) before, during, and after the test. The indices assessed were forced expiratory volume in one second, its ratio to the forced vital capacity, and peak expiratory flow rate. RESULTS: Mild to severe symptoms were experienced by 81% of patients during adenosine infusion. More than half the patients had dyspnea, but there was no associated bronchospasm. Although the patients with COPD showed parameters indicative of bronchial resistance compared with those without this problem, there was no further deterioration after adenosine infusion. Similarly, patients with a history of tobacco abuse, despite showing a tendency toward increasing airways resistance, had no bronchospasm during the administration of adenosine. CONCLUSIONS: (1) Although dyspnea is a common problem during adenosine stress MPI, it is not associated with any appreciable bronchospasm. (2) COPD and tobacco abuse do not appear to be contraindications to adenosine stress MPI studies. However, it is probably safe to exclude patients with severe COPD from adenosine stress MPI until after further evaluation with larger numbers of patients. PMID- 11431671 TI - Diagnosis of culture-negative endocarditis: the role of the Duke criteria and the impact of transesophageal echocardiography. AB - BACKGROUND: The Duke criteria have been shown to be more sensitive than the von Reyn criteria in the diagnosis of culture-positive endocarditis but to date have not been fully validated for culture-negative endocarditis (CNE). The aim of this study was (1) to compare the diagnostic accuracy of the Duke criteria versus clinical judgment and the von Reyn criteria in CNE and (2) to assess the diagnostic impact of transesophageal echocardiography (TEE) on the Duke criteria in CNE. METHODS: The study group consisted of 49 patients with suspected CNE in whom the presence (n = 32) or absence (n = 17) of endocarditis was confirmed by surgery, autopsy, or both. All patients underwent transthoracic echocardiography (TTE) and TEE. They were classified into a Duke category initially with TTE data only, and then the Duke categories were reevaluated with the additional TEE data. RESULTS: The Duke criteria demonstrated a significantly higher sensitivity (72%) than the von Reyn criteria (28%; P =.0008) and a higher specificity (100%) than clinical judgment (76%; P =.02). No major differences were noted between sensitivities of the Duke criteria and clinical judgement. TEE significantly augmented the capacity to diagnose CNE by Duke criteria versus TTE (P <.05). CONCLUSIONS: The Duke criteria are of high diagnostic validity for the conduction of clinical studies on CNE. They have the potential to affect clinical decision making, based on the higher specificity versus clinical judgment. TEE appears to be crucial for the diagnosis of CNE when the Duke criteria are applied. The diagnostic differentiation between CNE, sclerotic valve degeneration, and nonbacterial thrombotic endocarditis remains a challenge. PMID- 11431672 TI - Second harmonic imaging improves sensitivity of dobutamine stress echocardiography for the diagnosis of coronary artery disease. AB - OBJECTIVE: Our purpose was to assess the value of second harmonic imaging compared with fundamental imaging for the diagnosis of coronary artery disease during dobutamine stress echocardiography. PATIENTS AND METHODS: Sixty-four patients underwent dobutamine stress echocardiography with both fundamental imaging and second harmonic imaging. Coronary angiography was performed within 3 months. Ischemia was defined as new or worsening wall motion abnormalities in > or = 1 segment during dobutamine stress echocardiography. Coronary artery disease was defined as a > or = 70% luminal diameter stenosis in > or = 1 coronary artery by coronary angiography. RESULTS: There was a higher prevalence of segments with invisible border with fundamental compared with second harmonic imaging both at rest (11% vs 8%, P < .05) and at peak stress (17% vs 10%, P < .001). Significant coronary artery disease was present in 49 (77%) patients. The sensitivity of dobutamine stress echocardiography for detection of coronary artery disease by fundamental and second harmonic imaging was, respectively, 78% and 94% (P < .05), whereas specificity was similar (73% vs 73%). Second harmonic imaging had a particularly higher sensitivity for the diagnosis of 1-vessel disease (93% vs 50%, P < .05). CONCLUSION: The use of second harmonic imaging improves the sensitivity of dobutamine stress echocardiography for the diagnosis of coronary artery disease compared with fundamental imaging, particularly for 1-vessel coronary artery disease, whereas specificity remains unchanged. PMID- 11431673 TI - Cardiac rehabilitation and survival in patients with left ventricular systolic dysfunction. AB - BACKGROUND: Exercise training, the major component of cardiac rehabilitation (CR), has been shown in previous trials to improve many pathophysiologic changes found in patients with left ventricular systolic dysfunction. It remains unproven whether exercise training improves survival. METHODS: By using the Duke Databank for Cardiovascular Disease, we identified patients with an ejection fraction < or = 40% and no recent myocardial infarction, congenital heart disease, or primary valvular disease who survived > or = 30 days after a cardiac catheterization (n = 1902). Participation in CR (n = 70) was identified through computer billing records. We developed a multivariable Cox proportional hazards regression model to estimate survival by using variables known to be independent predictors of survival in patients with systolic dysfunction. RESULTS: Patients participating in CR were less likely to be female or black and more likely to have a history consistent with ischemic cardiomyopathy. Participation in CR was associated with significantly improved survival after adjustment for baseline characteristics (hazard ratio, 0.39; 95% confidence interval, 0.15 to 0.62, P < .0001). Survival increased when patients participated in > 6 CR sessions (hazard ratio, 0.10; 95% confidence interval, 0.03 to 0.39; P < .0001). CONCLUSIONS: Participation in CR was associated with improved survival for patients with cardiomyopathy. There appears to be a dose response with improved survival benefit for patients with left ventricular systolic dysfunction participating in cardiac rehabilitation. PMID- 11431674 TI - Relative lymphocyte count: a prognostic indicator of mortality in elderly patients with congestive heart failure. AB - BACKGROUND: A reduction in the relative lymphocyte count is a marker of the stress response; however, its prognostic value remains undetermined. The objective of this study was to investigate the predictive value of the relative lymphocyte count for survival in elderly patients with congestive heart failure (CHF). METHODS AND RESULTS: One thousand two hundred seventy-four consecutive patients above the age of 65 years hospitalized with heart disease were enrolled in the CHF Italian Study and followed up for 3 years. Of these, 413 patients were excluded because of factors that could affect the lymphocyte count. Of the remaining 861 patients, 423 (49%) met the criteria for the diagnosis of CHF (mean age 76 +/- 7 years, 51% men), of whom 162 patients (38%) had a relative lymphocyte count < or = 20%. The 3-year all-cause mortality in patients with CHF and a relative lymphocyte count < or = 20% was 64% compared with 40% in patients with a relative lymphocyte count > 20% (P < .0001). The age- and sex-adjusted hazard ratio for death in patients with CHF and low relative lymphocyte count was 1.76 (95% confidence interval 1.34-2.32, P = .0001). After adjustment for baseline differences and variables associated with or known to affect lymphocyte count, the hazard ratio remained significantly different from 1.0 (hazard ratio 1.73, 95% confidence interval 1.21-2.48, P = .0026). CONCLUSION: A low relative lymphocyte count is an independent marker of poor prognosis in elderly patients with CHF. The relative lymphocyte count is a simple, accurate, widely available, and inexpensive marker that can help to identify elderly patients with CHF who are at increased risk for mortality. The pathophysiologic mechanism of this observation remains to be determined. PMID- 11431675 TI - Chronic heart failure in the very elderly: clinical status, survival, and prognostic factors in 188 patients more than 70 years old. AB - BACKGROUND: Chronic heart failure (CHF) is a frequent disease with a dismal prognosis, but little is known about survival in the very elderly. There are no data on the prognostic value of cardiopulmonary exercise testing in this population. We aimed to assess exercise capacity, survival, and prognostic parameters in elderly patients with CHF. METHODS: We evaluated 188 patients with CHF >70 years old (mean 77 +/- 4 years, range 70-94 years) seen at our heart failure clinic between March 1992 and June 1998. A cardiopulmonary exercise test was performed in 102 patients (peak VO2 15.3 +/- 4.7, VE/VCO2 slope 39.6 +/- 15.01). All patients were followed up for at least 12 months. The prognostic end point of the study was all-cause mortality. RESULTS: At the end of follow-up (16 +/- 10 mo, range 12-41 mo), 67 patients (35.6%) had died (1-year mortality rate 26% [95% confidence interval 20-32]). In univariate analysis New York Heart Association class (NYHA) (relative risk [RR] = 2.56, P <.0001), VE/VCO2 (RR = 1.041, P <.0001), peak VO2 (RR = 0.87, P =.0007), and fractional shortening (RR = 0.95, P <.0001) predicted mortality. Peak VO2 predicted mortality independently of age, NYHA class, and left ventricular ejection fraction. A subgroup of 12 patients with dynamic left ventricular outflow tract obstruction during stress had an excellent outcome, with a 100% survival at the end of follow-up (mean 16 +/- 7 mo, range 12-39 mo). CONCLUSIONS: The prognosis in elderly patients with CHF is poor. Valid exercise testing results can be obtained in more than 50% of elderly patients with CHF. NYHA class and peak VO2 are the strongest prognostic factors in this population. PMID- 11431676 TI - Reduced right ventricular ejection fraction as a marker for idiopathic dilated cardiomyopathy compared with ischemic left ventricular dysfunction. AB - BACKGROUND: Evidence for the role of right ventricular (RV) function is emerging in patients with heart failure of different etiologies. Studies conducted in dilated cardiomyopathy (IDC) showed a high prevalence of RV dysfunction unrelated to the severity of pulmonary hypertension. The aim of the study was to investigate the role of RV dysfunction in ischemic versus nonischemic patients. METHODS: A series of 153 patients with left ventricular (LV) dysfunction (defined as a LV ejection fraction <45%) of either ischemic (n = 61, coronary artery disease [CAD] group) or nonischemic (n = 92, IDC group) origin were studied invasively. Besides routine catheterization data, RV volumes and ejection fractions were obtained angiographically. Reference data were collected in a control group of healthy subjects. RV dysfunction was defined as a RV ejection fraction <35% and ventricular concordance as a <10% difference between RV and LV ejection fraction. The LV/RV end-diastolic volume ratio was calculated to assess the relative dilatation of the ventricular chambers. Hemodynamic and angiographic data were compared in the 2 groups by univariate and multivariate logistic regression analysis. RESULTS: Patients with IDC and CAD had comparable LV ejection fractions (29% +/- 3% vs 31% +/- 8%, P not significant) and mean pulmonary pressures (27 +/- 12 mm Hg vs 26 +/- 11 mm Hg, P not significant); the LV/RV end-diastolic volume ratio was identical in the 2 groups (1.26 +/- 0.4 vs 1.24 +/- 0.4, P not significant). RV ejection fraction was significantly lower in IDC compared with CAD (33% +/- 10 % vs 46% +/- 11%, P <.0001), with a prevalence of RV dysfunction in the IDC group of 65% compared with 16% in the CAD group (P <.0001); similarly, the prevalence of ejection fraction concordance was 74% versus 33%, respectively (P <.0001). At multivariate analysis, a low RV ejection fraction was a powerful independent predictor of IDC compared with CAD (odds ratio 0.91, 95% confidence interval 0.87-0.94, P <.0001). RV dysfunction had a positive predictive value of 75% and a negative predictive value of 78% for the diagnosis of IDC; for ventricular concordance, these values were 81% and 69%, respectively. The correlation between mean pulmonary artery pressure and RV ejection fraction was weaker in the IDC group compared with the CAD group (R(2) = 0.032, P =.047 and R(2) = 0.172,P <.0001, respectively). CONCLUSION: In the presence of LV dysfunction, a reduced RV ejection fraction is a powerful marker for IDC compared with CAD, independent of age, pulmonary hypertension, LV function, and ventricular dimensions. These findings support the concept that IDC is frequently characterized by a biventricular involvement and that the presence of RV dysfunction represents a distinguishing feature of this disease. PMID- 11431677 TI - Urgent coronary bypass surgery for failed percutaneous coronary intervention in the stent era: Is backup still necessary? AB - BACKGROUND: Current practice guidelines for performance of percutaneous coronary intervention (PCI) in the United States mandate availability of on-site surgical backup. With the decreasing frequency of urgent coronary bypass surgery (UCABG) with newer technologies, it is unclear whether such backup continues to be necessary. METHODS: A database of 5655 consecutive patients undergoing PCI at a single center between August 1, 1992, and December 31, 1997, was analyzed. Outcomes were determined as well as clinical, lesion, and procedural characteristics of patients during 4 time periods preceding and during use of coronary stenting. RESULTS: Frequency of UCABG for failed PCI decreased from 2.2% to 0.6% in the most recent time period (P <.01) with no change in incidence of in hospital death or myocardial infarction. Incidence of stenting progressively increased to 72% in the latest period. Patients requiring UCABG had a higher prevalence of acute coronary syndromes (95%) and type B lesions (79%), but these characteristics were also common in patients who did not undergo UCABG. Although coronary stents were available during the last 3 periods studied, only 30% of UCABG patients had lesions or complications amenable to stenting, and stenting attempts in these patients were all unsuccessful. Despite stenting and use of perfusion balloons and intra-aortic balloon pumps, only 40% of patients having UCABG were stable and pain free on transfer to the operating room. CONCLUSIONS: Although use of UCABG for a failed PCI is currently very low, there are no satisfactory predictors, patients requiring UCABG are frequently clinically unstable, and availability of stenting does not reliably eliminate the need for UCABG or result in a decrease in mortality. This small group of patients continues to require readily available surgical standby. PMID- 11431678 TI - Protecting individuals and promoting science. PMID- 11431679 TI - The history of life. PMID- 11431680 TI - Mouse mutagenesis on target. AB - Large-scale mutagenesis of the mouse genome is an essential task associated with the Human Genome Project. The two opposing schools of direct and reverse genetics have demonstrated comparable advantages, and yet large numbers of mutant lines have mostly been the prerogative of direct genetics. An improved gene-trapping resource now brings reverse genetics one step closer. PMID- 11431681 TI - A helicase is born. AB - One of three loci previously associated with autosomal dominant progressive external ophthalmoplegia (adPEO) encodes ANT1, a mitochondrial nucleotide transporter. Now, mutations in two other genes are found in people with adPEO. One of these encodes a new helicase, Twinkle, which is related to the product of bacteriophage T7 gene 4, and co-localizes with mitochondrial DNA. The identification of Twinkle adds a new star to the expanding constellation of 'helicase diseases'. PMID- 11431682 TI - Don't mess with the matrix. AB - Extracellular matrix (ECM) remodeling is critical to morphogenesis and homeostasis. The identification of inactivating mutations in a gene encoding one of its modifying enzymes, matrix metalloproteinase 2 (MMP-2), in people with a hereditary disorder in which the bones disintegrate, represents the first genetic evidence that the proteolysis of the ECM mediates human growth and development. It also underscores the need for an intricate balance between breakdown and deposition of the ECM. PMID- 11431683 TI - Deconstructing maize population structure. AB - Association studies have rarely been used in plant genetics, in part because of the risk of false positives caused by population structure. A study of flowering time in maize makes the first use of recent 'structured association' methods statistical approaches that use independent loci to control for the effects of structure and admixture. PMID- 11431685 TI - Consent and privacy in pharmacogenetic testing. AB - The clinical use of pharmacogenetic drugs will require that a sample of a patient's DNA be tested before a drug is prescribed. Although pharmacogenetic tests pose fewer risks than genetic tests for disease mutations, they might still reveal personal information that could be used adversely to a patient's interests. Informed consent and privacy of pharmacogenetic test results may be essential in most clinical uses of pharmacogenetic drugs. PMID- 11431686 TI - Mutation of POLG is associated with progressive external ophthalmoplegia characterized by mtDNA deletions. AB - Progressive external ophthalmoplegias (PEO) characterized by accumulation of large-scale mitochondrial DNA (mtDNA) deletions are rare human diseases. We mapped a new locus for dominant PEO at 15q22-q26 in a Belgian pedigree and identified a heterozygous mutation (Y955C) in the polymerase motif B of the mtDNA polymerase gamma (POLG). We identified three additional POLG missense mutations compatible with recessive PEO In two nuclear families. POLG is the only DNA polymerase responsible for mtDNA replication. PMID- 11431687 TI - A mutation in SLC11A3 is associated with autosomal dominant hemochromatosis. AB - Hereditary hemochromatosis (HH) is a very common disorder characterized by iron overload and multi-organ damage. Several genes involved in iron metabolism have been implicated in the pathology of HH (refs. 1-4). We report that a mutation in the gene encoding Solute Carrier family 11, member A3 (SLC11A3), also known as ferroportin, is associated with autosomal dominant hemochromatosis. PMID- 11431688 TI - PAX6 haploinsufficiency causes cerebral malformation and olfactory dysfunction in humans. AB - PAX6 is widely expressed in the central nervous system. Heterozygous PAX6 mutations in human aniridia cause defects that would seem to be confined to the eye. Magnetic resonance imaging (MRI) and smell testing reveal the absence or hypoplasia of the anterior commissure and reduced olfaction in a large proportion of aniridia cases, which shows that PAX6 haploinsuffiency causes more widespread human neuro developmental anomalies. PMID- 11431689 TI - Sox9 induces testis development in XX transgenic mice. AB - Mutations in SOX9 are associated with male-to-female sex reversal in humans. To analyze Sox9 function during sex determination, we ectopically expressed this gene in XX gonads. Here, we show that Sox9 is sufficient to induce testis formation in mice, indicating that it can substitute for the sex-determining gene Sry. PMID- 11431690 TI - Mutations in CAV3 cause mechanical hyperirritability of skeletal muscle in rippling muscle disease. AB - Hereditary rippling muscle disease (RMD) is an autosomal dominant human disorder characterized by mechanically triggered contractions of skeletal muscle. Genome wide linkage analysis has identified an RMD locus on chromosome 3p25. We found missense mutations in positional candidate CAV3 (encoding caveolin 3; ref. 5) in all five families analyzed. Mutations in CAV3 have also been described in limb girdle muscular dystrophy type 1C (LGMD1C; refs. 6,7), demonstrating the allelism of dystrophic and non-dystrophic muscle diseases. PMID- 11431691 TI - Fusion of two novel genes, RBM15 and MKL1, in the t(1;22)(p13;q13) of acute megakaryoblastic leukemia. AB - t(1;22) is the principal translocation of acute megakaryoblastic leukemias. Here we show this chromosomal rearrangement to result in the fusion of two novel genes, RNA-binding motif protein-15 (RBM15), an RNA recognition motif-encoding gene with homology to Drosophila spen, and Megakaryoblastic Leukemia-1 (MKL1), a gene encoding an SAP (SAF-A/B, Acinus and PIAS) DNA-binding domain. PMID- 11431692 TI - Human mitochondrial DNA deletions associated with mutations in the gene encoding Twinkle, a phage T7 gene 4-like protein localized in mitochondria. AB - The gene products involved in mammalian mitochondrial DNA (mtDNA) maintenance and organization remain largely unknown. We report here a novel mitochondrial protein, Twinkle, with structural similarity to phage T7 gene 4 primase/helicase and other hexameric ring helicases. Twinkle colocalizes with mtDNA in mitochondrial nucleoids. Screening of the gene encoding Twinkle in individuals with autosomal dominant progressive external ophthalmoplegia (adPEO), associated with multiple mtDNA deletions, identified 11 different coding-region mutations co segregating with the disorder in 12 adPEO pedigrees of various ethnic origins. The mutations cluster in a region of the protein proposed to be involved in subunit interactions. The function of Twinkle is inferred to be critical for lifetime maintenance of human mtDNA integrity. PMID- 11431693 TI - The SNRPN promoter is not required for genomic imprinting of the Prader Willi/Angelman domain in mice. AB - In mice and humans, the locus encoding the gene for small nuclear ribonucleoprotein N (SNRPN/Snrpn), as well as other loci in the region are subject to genomic imprinting. The SNRPN promoter is embedded in a maternally methylated CpG island, is expressed only from the paternal chromosome and lies within an imprinting center that is required for switching to and/or maintenance of the paternal epigenotype. We show here that a 0.9-kb deletion of exon 1 of mouse Snrpn did not disrupt imprinting or elicit any obvious phenotype, although it did allow the detection of previously unknown upstream exons. In contrast, a larger, overlapping 4.8-kb deletion caused a partial or mosaic imprinting defect and perinatal lethality when paternally inherited. PMID- 11431694 TI - Functional analysis of secreted and transmembrane proteins critical to mouse development. AB - We describe the successful application of a modified gene-trap approach, the secretory trap, to systematically analyze the functions in vivo of large numbers of genes encoding secreted and membrane proteins. Secretory-trap insertions in embryonic stem cells can be transmitted to the germ line of mice with high efficiency and effectively mutate the target gene. Of 60 insertions analyzed in mice, one-third cause recessive lethal phenotypes affecting various stages of embryonic and postnatal development. Thus, secretory-trap mutagenesis can be used for a genome-wide functional analysis of cell signaling pathways that are critical for normal mammalian development and physiology. PMID- 11431695 TI - Ltap, a mammalian homolog of Drosophila Strabismus/Van Gogh, is altered in the mouse neural tube mutant Loop-tail. AB - Neural tube defects (NTDs) such as spina bifida and anencephaly are common congenital malformations in humans (1/1,000 births) that result from failure of the neural tube to close during embryogenesis. The etiology of NTDs is complex, with both genetic and environmental contributions; the genetic component has been extensively studied with mouse models. Loop-tail (Lp) is a semidominant mutation on mouse chromosome 1 (ref. 4). In the two known Lp alleles (Lp, Lpm1Jus), heterozygous mice exhibit a characteristic looped tail, and homozygous embryos show a completely open neural tube in the hindbrain and spinal region, a condition similar to the severe craniorachischisis defect in humans. Morphological and neural patterning studies indicate a role for the Lp gene product in controlling early morphogenesis and patterning of both axial midline structures and the developing neural plate. The 0.6-cM/0.7-megabase (Mb) Lp interval is delineated proximally by D1Mit113/Apoa2/Fcer1g and distally by Fcer1a/D1Mit149/Spna1 and contains a minimum of 17 transcription units. One of these genes, Ltap, encodes a homolog of Drosophila Strabismus/Van Gogh (Stbm/Vang), a component of the frizzled/dishevelled tissue polarity pathway. Ltap is expressed broadly in the neuroectoderm throughout early neurogenesis and is altered in two independent Lp alleles, identifying this gene as a strong candidate for Lp. PMID- 11431696 TI - A photic visual cycle of rhodopsin regeneration is dependent on Rgr. AB - During visual excitation, rhodopsin undergoes photoactivation and bleaches to opsin and all-trans-retinal. To regenerate rhodopsin and maintain normal visual sensitivity, the all-trans isomer must be metabolized and reisomerized to produce the chromophore 11-cis-retinal in biochemical steps that constitute the visual cycle and involve the retinal pigment epithelium (RPE; refs. 3-8). A key step in the visual cycle is isomerization of an all-trans retinoid to 11-cis-retinol in the RPE (refs. 9-11). It could be that the retinochrome-like opsins, peropsin, or the retinal G protein-coupled receptor (RGR) opsin12-16 are isomerases in the RPE. In contrast to visual pigments, RGR is bound predominantly to endogenous all trans-retinal, and irradiation of RGR in vitro results in stereospecific conversion of the bound all-trans isomer to 11-cis-retinal. Here we show that RGR is involved in the formation of 11-cis-retinal in mice and functions in a light dependent pathway of the rod visual cycle. Mutations in the human gene encoding RGR are associated with retinitis pigmentosa. PMID- 11431697 TI - Mutation of the matrix metalloproteinase 2 gene (MMP2) causes a multicentric osteolysis and arthritis syndrome. AB - The inherited osteolyses or 'vanishing bone' syndromes are a group of rare disorders of unknown etiology characterized by destruction and resorption of affected bones. The multicentric osteolyses are notable for interphalangeal joint erosions that mimic severe juvenile rheumatoid arthritis (OMIMs 166300, 259600, 259610 and 277950). We recently described an autosomal recessive form of multicentric osteolysis with carpal and tarsal resorption, crippling arthritic changes, marked osteoporosis, palmar and plantar subcutaneous nodules and distinctive facies in a number of consanguineous Saudi Arabian families. We localized the disease gene to 16q12-21 by using members of these families for a genome-wide search for homozygous-by-descent microsatellite markers. Haplotype analysis narrowed the critical region to a 1.2-cM region that spans the gene encoding MMP-2 (gelatinase A, collagenase type IV; (ref. 3). We detected no MMP2 enzymatic activity in the serum or fibroblasts of affected family members. We identified two family-specific homoallelic MMP2 mutations: R101H and Y244X. The nonsense mutation effects a deletion of the substrate-binding and catalytic sites and the fibronectin type II-like and hemopexin/TIMP2 binding domains. Based on molecular modeling, the missense mutation disrupts hydrogen bond formation within the highly conserved prodomain adjacent to the catalytic zinc ion. PMID- 11431698 TI - Genetic interactions between tumor suppressors Brca1 and p53 in apoptosis, cell cycle and tumorigenesis. AB - Breast cancer is a chief cause of cancer-related mortality that affects women worldwide. About 8% of cases are hereditary, and approximately half of these are associated with germline mutations of the breast tumor suppressor gene BRCA1 (refs. 1,2). We have previously reported a mouse model in which Brca1 exon 11 is eliminated in mammary epithelial cells through Cre-mediated excision. This mutation is often accompanied by alterations in transformation-related protein 53 (Trp53, encoding p53), which substantially accelerates mammary tumor formation. Here, we sought to elucidate the underlying mechanism(s) using mice deficient in the Brca1 exon 11 isoform (Brca1Delta11/Delta11). Brca1Delta11/Delta11 embryos died late in gestation because of widespread apoptosis. Unexpectedly, elimination of one Trp53 allele completely rescues this embryonic lethality and restores normal mammary gland development. However, most female Brca1Delta11/Delta11 Trp53+/- mice develop mammary tumors with loss of the remaining Trp53 allele within 6-12 months. Lymphoma and ovarian tumors also occur at lower frequencies. Heterozygous mutation of Trp53 decreases p53 and results in attenuated apoptosis and G1-S checkpoint control, allowing Brca1Delta11/Delta11 cells to proliferate. The p53 protein regulates Brca1 transcription both in vitro and in vivo, and Brca1 participates in p53 accumulation after gamma-irradiation through regulation of its phosphorylation and Mdm2 expression. These findings provide a mechanism for BRCA1-associated breast carcinogenesis. PMID- 11431699 TI - Human cells are protected from mitochondrial dysfunction by complementation of DNA products in fused mitochondria. AB - Extensive complementation between fused mitochondria is indicated by recombination of 'parental' mitochondrial (mt) DNA (ref. 1,2) of yeast and plant cells. It has been difficult, however, to demonstrate the occurrence of complementation between fused mitochondria in mammalian species through the presence of recombinant mtDNA molecules, because sequence of mtDNA throughout an individual tends to be uniform owing to its strictly maternal inheritance. We isolated two types of respiration-deficient cell lines, with pathogenic mutations in mitochondrial tRNAIle or tRNALeu(UUR) genes from patients with mitochondrial diseases. The coexistence of their mitochondria within hybrids restored their normal morphology and respiratory enzyme activity by 10-14 days after fusion, indicating the presence of an extensive and continuous exchange of genetic contents between the mitochondria. This complementation between fused mitochondria may represent a defence of highly oxidative organelles against mitochondrial dysfunction caused by the accumulation of mtDNA lesions with age. PMID- 11431700 TI - Tbx5 associates with Nkx2-5 and synergistically promotes cardiomyocyte differentiation. AB - The cardiac homeobox protein Nkx2-5 is essential in cardiac development, and mutations in Csx (which encodes Nkx2-5) cause various congenital heart diseases. Using the yeast two-hybrid system with Nkx2-5 as the 'bait', we isolated the T box-containing transcription factor Tbx5; mutations in TBX5 cause heart and limb malformations in Holt-Oram syndrome (HOS). Co-transfection of Nkx2-5 and Tbx5 into COS-7 cells showed that they also associate with each other in mammalian cells. Glutathione S-transferase (GST) 'pull-down' assays indicated that the N terminal domain and N-terminal part of the T-box of Tbx5 and the homeodomain of Nkx2-5 were necessary for their interaction. Tbx5 and Nkx2-5 directly bound to the promoter of the gene for cardiac-specific natriuretic peptide precursor type A (Nppa) in tandem, and both transcription factors showed synergistic activation. Deletion analysis showed that both the N-terminal domain and T-box of Tbx5 were important for this transactivation. A G80R mutation of Tbx5, which causes substantial cardiac defects with minor skeletal abnormalities in HOS, did not activate Nppa or show synergistic activation, whereas R237Q, which causes upper limb malformations without cardiac abnormalities, activated the Nppa promoter to a similar extent to that of wildtype Tbx5. P19CL6 cell lines overexpressing wildtype Tbx5 started to beat earlier and expressed cardiac-specific genes more abundantly than did parental P19CL6 cells, whereas cell lines expressing the G80R mutant did not differentiate into beating cardiomyocytes. These results indicate that two different types of cardiac transcription factors synergistically induce cardiac development. PMID- 11431701 TI - Universal trees based on large combined protein sequence data sets. AB - Universal trees of life based on small-subunit (SSU) ribosomal RNA (rRNA) support the separate mono/holophyly of the domains Archaea (archaebacteria), Bacteria (eubacteria) and Eucarya (eukaryotes) and the placement of extreme thermophiles at the base of the Bacteria. The concept of universal tree reconstruction recently has been upset by protein trees that show intermixing of species from different domains. Such tree topologies have been attributed to either extensive horizontal gene transfer or degradation of phylogenetic signals because of saturation for amino acid substitutions. Here we use large combined alignments of 23 orthologous proteins conserved across 45 species from all domains to construct highly robust universal trees. Although individual protein trees are variable in their support of domain integrity, trees based on combined protein data sets strongly support separate monophyletic domains. Within the Bacteria, we placed spirochaetes as the earliest derived bacterial group. However, elimination from the combined protein alignment of nine protein data sets, which were likely candidates for horizontal gene transfer, resulted in trees showing thermophiles as the earliest evolved bacterial lineage. Thus, combined protein universal trees are highly congruent with SSU rRNA trees in their strong support for the separate monophyly of domains as well as the early evolution of thermophilic Bacteria. PMID- 11431702 TI - Dwarf8 polymorphisms associate with variation in flowering time. AB - Historically, association tests have been used extensively in medical genetics, but have had virtually no application in plant genetics. One obstacle to their application is the structured populations often found in crop plants, which may lead to nonfunctional, spurious associations. In this study, statistical methods to account for population structure were extended for use with quantitative variation and applied to our evaluation of maize flowering time. Mutagenesis and quantitative trait locus (QTL) studies suggested that the maize gene Dwarf8 might affect the quantitative variation of maize flowering time and plant height. The wheat orthologs of this gene contributed to the increased yields seen in the 'Green Revolution' varieties. We used association approaches to evaluate Dwarf8 sequence polymorphisms from 92 maize inbred lines. Population structure was estimated using a Bayesian analysis of 141 simple sequence repeat (SSR) loci. Our results indicate that a suite of polymorphisms associate with differences in flowering time, which include a deletion that may alter a key domain in the coding region. The distribution of nonsynonymous polymorphisms suggests that Dwarf8 has been a target of selection. PMID- 11431703 TI - Regulation of premeiotic S phase and recombination-related double-strand DNA breaks during meiosis in fission yeast. AB - The meiotic cell cycle is characterized by high levels of recombination induced by DNA double-strand breaks (DSBs), which appear after completion of premeiotic S phase, leading to the view that initiation of recombination depends on meiotic DNA replication. It has also been indicated that DNA replication initiation proteins may differ between the meiotic and mitotic cell cycles, giving rise to an altered S phase, which could contribute to the high level of recombination during meiosis. We have investigated these possibilities in the fission yeast Schizosaccharomyces pombe and found that core DNA replication initiation proteins used during the mitotic cell cycle, including Cdc18p (budding yeast Cdc6p), Cdc19p (Mcm2p), Cdc21p (Mcm4p) and Orp1p (Orc1p), are also required for premeiotic S phase. Reduced activity of these proteins prevents completion of DNA replication but not formation of DSBs. We conclude that recombination-related DSB formation does not depend on the completion of meiotic DNA replication and we propose two parallel developmental sequences during the meiotic cell cycle: one for premeiotic S phase and the other for initiating recombination. PMID- 11431704 TI - Mitotic recombination effects homozygosity for NF1 germline mutations in neurofibromas. AB - Pure populations of neurofibroma-derived Schwann cells bearing both NF1 mutated alleles (NF1-/-) have been isolated from different neurofibromas showing loss of heterozygosity of nearly the entire 17q chromosome. By comparing molecular and fluorescent in situ hybridization analysis of these cells, we demonstrate mitotic recombination is the mechanism underlying this type of loss of heterozygosity leading to reduction to homozygosity of NF1 germline mutation. PMID- 11431705 TI - Unusual haplotypic structure of IL8, a susceptibility locus for a common respiratory virus. AB - Interleukin-8 (IL8) is believed to play a role in the pathogenesis of bronchiolitis, a common viral disease of infancy, and a recent U.K. family study identified an association between this disease and the IL8-251A allele. In the present study we report data, from a different set of families, which replicate this finding; combined analysis of 194 nuclear families through use of the transmission/disequilibrium test gives P = .001. To explore the underlying genetic cause, we identified nine single-nucleotide polymorphisms (SNPs) in a 7.6 kb segment spanning the IL8 gene and its promoter region and used six of these SNPs to define the haplotypic structure of the IL8 locus. The IL8-251A allele resides on two haplotypes, only one of which is associated with disease, suggesting that this may not be the functional allele. Europeans show an unusual haplotype genealogy that is dominated by two common haplotypes differing at multiple sites, whereas Africans have much greater haplotypic diversity. These marked haplotype-frequency differences give an F(ST) of.25, and, in the European sample, both Tajima's D statistic (D = 2.58, P = .007) and the Hudson/Kreitman/Aguade test (chi(2) = 4.9, P = .03) reject neutral equilibrium, suggesting that selective pressure may have acted on this locus. PMID- 11431706 TI - A novel frameshift mutation in exon 23 of ATP7A (MNK) results in occipital horn syndrome and not in Menkes disease. AB - Menkes disease and occipital horn syndrome (OHS) are allelic, X-linked recessive copper-deficiency disorders resulting from mutations in ATP7A, or MNK. Classic Menkes disease has a severe phenotype, with death in early childhood, whereas OHS has a milder phenotype, with, mainly, connective-tissue abnormalities. Data suggest that steady-state localization of ATP7A to the trans-Golgi network (TGN) is necessary for proper activity of lysyl oxidase, which is the predominant cuproenzyme whose activity is deficient in OHS and which is essential for maintenance of connective-tissue integrity. Recently, it was reported that ATP7A transcript levels as low as 2%-5% of normal are sufficient to result in the milder phenotype, OHS, rather than the phenotype of Menkes disease. In contrast to previously reported cases of OHS, we describe a case of OHS in which, because of a frameshift mutation, no normal ATP7A is produced. Although abundant levels of mutant transcript are present, there are substantially reduced levels of the truncated protein, which lacks the key dileucine motif L1487L1488. It has been demonstrated that the dileucine motif L1487L1488 functions as an endocytic signal for ATP7A cycling between the TGN and the plasma membrane. The present report is the first to describe an ATP7A truncation that results in OHS rather than in Menkes disease. The data from the present report support the concepts that (1) OHS results from lower levels of functional ATP7A and (2) ATP7A does not require the dileucine motif to function in copper efflux. PMID- 11431707 TI - Estimating the efficacy and efficiency of cascade genetic screening. AB - Screening for genetic variants that predispose individuals or their offspring to disease may be performed at the general population level or may instead be targeted at the relatives of previously identified carriers. The latter strategy has come to be known as "cascade genetic screening." Since the carrier risk of close relatives of known carriers is generally higher than the population risk, cascade screening is more efficient than population screening, in the sense that fewer individuals have to be genotyped per detected carrier. The efficacy of cascade screening, as measured by the overall proportion of carriers detected in a given population, is, however, lower than that of population-wide screening, and the respective inclusion rates vary according to the population frequency and mode of inheritance of the predisposing variants. For dominant mutations, we have developed equations that allow the inclusion rates of cascade screening to be calculated in an iterative fashion, depending upon screening depth and penetrance. For recessive mutations, we derived only equations for the screening of siblings and the children of patients. Owing to their mathematical complexity, it was necessary to study more extended screening strategies by simulation. Cascade screening turned out to result in low inclusion rates (<1%) when aimed at the identification of heterozygous carriers of rare recessive variants. Considerably higher rates are achievable, however, when screening is performed to detect covert homozygotes for frequent recessive mutations with reduced penetrance. This situation is exemplified by hereditary hemochromatosis, for which up to 40% of at-risk individuals may be identifiable through screening of first- to third-degree relatives of overt carriers (i.e., patients); the efficiency of this screening strategy was found to be approximately 50 times higher than that of population-wide screening. For dominant mutations, inclusion rates of cascade screening were estimated to be higher than for recessive variants. Thus, some 80% of all carriers of the factor V Leiden mutation would be detected if screening were to be targeted specifically at first- to third-degree relatives of patients with venous thrombosis. The relative cost efficiency of cascade as compared with population-wide screening (i.e., the overall savings in the extra managerial cost of the condition) is also likely to be higher for dominant than for recessive mutations. This notwithstanding, once screening has become cost-effective at the population level, it can be expected that cascade screening would only transiently represent an economically viable option. PMID- 11431708 TI - Disruption of the ProSAP2 gene in a t(12;22)(q24.1;q13.3) is associated with the 22q13.3 deletion syndrome. AB - The terminal 22q13.3 deletion syndrome is characterized by severe expressive language delay, mild mental retardation, hypotonia, joint laxity, dolichocephaly, and minor facial dysmorphisms. We identified a child with all the features of 22q13.3 deletion syndrome. The patient's karyotype showed a de novo balanced translocation between chromosomes 12 and 22, with the breakpoint in the 22q13.3 critical region of the 22q distal deletion syndrome [46, XY, t(12;22)(q24.1;q13.3)]. FISH investigations revealed that the translocation was reciprocal, with the chromosome 22 breakpoint within the 22q subtelomeric cosmid 106G1220 and the chromosome 12q breakpoint near STS D12S317. Using Southern blot analysis and inverse PCR, we located the chromosome 12 breakpoint in an intron of the FLJ10659 gene and located the chromosome 22 breakpoint within exon 21 of the human homologue of the ProSAP2 gene. Short homologous sequences (5-bp, CTG[C/A]C) were found at the breakpoint on both derivative chromosomes. The translocation does not lead to the loss of any portion of DNA. Northern blot analysis of human tissues, using the rat ProSAP2 cDNA, showed that full-length transcripts were found only in the cerebral cortex and the cerebellum. The FLJ10659 gene is expressed in various tissues and does not show tissue-specific isoforms. The finding that ProSAP2 is included in the critical region of the 22q deletion syndrome and that our proband displays all signs and symptoms of the syndrome suggests that ProSAP2 haploinsufficiency is the cause of the 22q13.3 deletion syndrome. ProSAP2 is a good candidate for this syndrome, because it is preferentially expressed in the cerebral cortex and the cerebellum and encodes a scaffold protein involved in the postsynaptic density of excitatory synapses. PMID- 11431709 TI - Molecular basis of disease processes. PMID- 11431710 TI - Molecular pathology of endometrial hyperplasia and carcinoma. AB - Four different genetic abnormalities may occur in endometrioid adenocarcinomas of the endometrium (mircosatellite instability and mutations in the PTEN, k-RAS and beta-catenin genes), whereas nonendometrioid carcinomas of the endometrium often have p53 mutations and loss of heterozygosity on several chromosomes. Occasionally, a nonendometrioid carcinoma may develop as a result of dedifferentiation of a preexisting endometrioid carcinoma; in such a case, the tumor exhibits overlapping clinical, morphologic, immunohistochemical, and molecular features of the 2 types. The insaturation of microsatellite instability in endometrial carcinogenesis seems to occur late in the transition from complex hyperplasia to carcinoma, and it is preceded by progressive inactivation of MLH-1 by promoter hypermethylation. Moreover, the endometrioid adenocarcinomas that exhibit microsatellite instability show a stepwise progressive accumulation of secondary mutations in oncogenes and tumor suppressor genes that contain short tandem repeats in their coding sequences. Mutations in the PTEN and k-RAS genes are also frequent in endometrioid adenocarcinomas of the endometrium, particularly in the tumors that exhibit microsatellite instability, whereas beta catenin mutations do not seem to be associated with such a phenomenon. PMID- 11431711 TI - Gastrointestinal stromal tumor workshop. AB - Gastrointestinal stromal tumor (GIST) has emerged in the past year as a prototypical neoplasm that responds to therapy directed against a single target molecule-the KIT receptor tyrosine kinase protein. Although GIST seldom responds to conventional chemotherapeutic agents, early experience with the tyrosine kinase inhibitor, STI-571 (Gleevec; Novartis, Basel, Switzerland), has been extremely encouraging. Early results have appeared in a recent case report in the New England Journal of Medicine (April 5, 2001),(1) and in early clinical trials from the United States and Europe that were reported at the plenary session of the American Society of Clinical Oncology in San Francisco on May 14, 2001. STI 571 is one of the earliest examples of a nontoxic chemotherapeutic agent (an agent whose anti-cancer activity is not predicated on a cytotoxic mechanism). STI 571 has already shown clinical value in BCR-ABL-positive leukemias. Early clinical results in GIST are so encouraging that oncologists may soon be wrestling with the opportunity of referring every patient with malignant GIST into clinical trials with STI-571. To ensure appropriate treatment, pathologists need to understand the biology and treatment of this tumor and to have standard methods and criteria for providing diagnosis (GIST or not GIST) and consistent prognostic classification (high risk of metastasis or low risk of metastasis). PMID- 11431712 TI - Invasive micropapillary carcinoma of the breast: eighty cases of an underrecognized entity. AB - Eighty cases of infiltrating duct carcinoma of the breast with a pure or partial micropapillary component are reported. The cases were analyzed using various parameters, including age at presentation, tumor size, tumor grade, presence of lymphatic invasion, and axillary lymph node status. The patients' age at presentation ranged from 36 to 92 years (mean, 58.8 years). Tumor size ranged from 0.1 to 10 cm (mean, 2.0 cm); 67.5% (54 of 80) were poorly differentiated, and 32.5% (26 of 80) were moderately differentiated; 62.5% (50 of 80) of the cases had lymphatic invasion; 72.3% (47 of 65) of cases with axillary dissections had positive lymph nodes; and 25% (20 of 80) of the tumors were < or = 1.0 cm and 7.5% (6 of 80) were < or = 0.5 cm. The characteristics of these small tumors with an invasive micropapillary component have not previously been reported. Despite their minute size, these small micropapillary carcinomas seem to show the same proclivity for lymphatic spread and nodal dissemination as larger tumors with this unique histologic pattern. PMID- 11431713 TI - Morphologic typing of papillary renal cell carcinoma: comparison of growth kinetics and patient survival in 66 cases. AB - Whereas papillary renal cell carcinoma is now established as a subtype of renal cell neoplasia, division of these tumors into 2 distinctive morphotypes has been proposed. Type 1 tumors have cells with scanty pale cytoplasm arranged in a single layer on the basement membrane of papillary cores. In these tumors, psammoma bodies and foamy macrophages are frequently seen, and the tumors frequently express cytokeratin 7. Type 2 tumor cells have pseudostratified nuclei and usually have voluminous eosinophilic cytoplasm. Recent studies have supported this subclassification of papillary renal cell carcinoma by demonstrating differing genotypes for type 1 and 2 tumors. To further study the subclassification of papillary renal carcinoma, we compared clinical features, nuclear grade, stage, tumor growth kinetics, and survival in a series of 50 type 1 and 16 type 2 papillary renal cell carcinomas. Comparison of patient age at presentation, sex, and primary tumor size shows no significant difference between the 2 tumor types. Type 1 tumors were of significantly lower Fuhrman grade (P =.0001) and higher Robson stage (P =.009) than type 2 tumors. There was no significant difference when tumors were staged according to the TNM classification. Assessment of tumor growth kinetics showed significantly different mean silver-staining nucleolar organizer region (AgNOR) scores and Ki 67 indices (AgNOR type 1, 3.83, type 2, 7.24, P =.0001; Ki-67 type 1, 3.17%, type 2, 6.01%, P =.0002). Multivariate analysis showed tumor type (P =.03), presence of metastases (P =.04), AgNOR score (P =.001), and Ki-67 index (P =.03) to be independently associated with survival. These results provide evidence of the clinical utility of dividing papillary renal cell carcinomas into 2 types according to histologic characteristics. PMID- 11431714 TI - Carcinoma ex pleomorphic adenoma: pathologic analysis of 73 cases. AB - Pathologic factors of predictive value for carcinoma ex pleomorphic adenoma (CXPA), an aggressive salivary gland malignancy, are poorly defined. Because residual mixed tumor may be relatively inconspicuous and various carcinoma subtypes are encountered, misdiagnosis is common. To describe the pathologic features and identify potential prognostic factors, we retrospectively examined 73 cases of CXPA of the major salivary glands treated at Mayo Clinic. Paraffin section immunostaining for keratins (AE1/AE3, CK7, CK20), epithelial membrane antigen, carcinoembryonic antigen, vimentin, actin, S-100 protein, glial fibrillary acidic protein, and p53 and c-erbB-2 oncoproteins was performed in 69 cases. DNA content and proliferation indices were determined by digital image analysis of Feulgen- and MIB-I-stained sections, retrospectively. Survival was calculated by the Kaplan-Meier method, and prognostic variables were analyzed with the log-rank test. The carcinoma component was predominant in 82% of tumors. Adenocarcinoma not otherwise specified (31 cases) and salivary duct carcinoma (24 cases) were the most frequent histologic subtypes. Sixty-two tumors were high grade (Broders 3 or 4). Residual mixed tumor was extensively hyalinized in 54 cases. Pathologic features significantly associated with overall survival included pathologic stage (P =.009), tumor size (P =.012), grade (P =.005), proportion of carcinoma (P =.004), extent of invasion (P =.002), and proliferation index of carcinoma (P =.03). Of 4 patients with intracapsular (noninvasive) carcinoma, none had an adverse outcome. The immunohistochemical profile of CXPA included positive staining reactions in the malignant component for AE1/AE3 in 97% of cases, CK7 in 94%, epithelial membrane antigen in 86%, carcinoembryonic antigen in 75%, vimentin in 52%, and S-100 protein in 29%. Expression of p53 and c-erbB-2 oncoproteins was detected in 41% and 30% of the carcinomas, respectively, but neither was associated with decreased survival. High-grade salivary adenocarcinoma that is difficult to classify should raise the suspicion of possible CXPA. Intracapsular carcinoma has a benign clinical course. Significant prognostic factors in CXPA include tumor stage, grade, proportion of carcinoma, extent of invasion, and proliferation index. PMID- 11431715 TI - High telomerase activity is associated with cell cycle deregulation and rapid progression in endometrioid adenocarcinoma of the uterus. AB - Telomerase activity, a mechanism granting cellular immortality, has been detected in most cancer entities, but its association with clinical, histopathologic, and prognostic parameters is not fully understood. We investigated whether quantitative telomerase levels are correlated to established prognostic factors, telomere lengths, cell cycle kinetics, and the clinical course in endometrioid adenocarcinoma of the uterus (EC). A modified telomeric repeat amplification protocol (TRAP) was used to quantify the relative telomerase activity in a series of 53 primary tumors. Mean telomere length was determined by Southern blot analysis. Cell cycle kinetics were studied immunohistochemically on paraffin sections using monoclonal antibodies to 2 distinct proliferation-specific proteins: Ki-67, which is expressed throughout the cell cycle, and a novel cell cycle-associated protein, repp86, the expression of which is restricted to the cell cycle phases S, G2, and M. The ratio of the 2 immunolabeling indices defines the rate of transition through the restriction point. Telomerase activity was detected in 50 of 53 ECs (94%). Its levels correlated significantly with FIGO stage (P =.01) and FIGO grade (P =.003) but not with myometrial invasion. They were weakly associated with the overall proliferative activity (Ki-67, r =.48) but significantly with the repp86 index (r =.64) and even more strongly with the repp86:Ki-67 ratio (r =.77). There was no correlation with mean telomere length. In the group of tumors with high telomerase activity, 5 patients had relapses and 2 died of the disease within a median follow-up period of 29 months. Recurrence showed no relation to FIGO grade and stage. No events were observed in the group with low telomerase activity. In a multivariate model including tumor stage, histopathologic grade, depth of myometrial invasion, and Ki-67 indices, telomerase activity emerged as the only independent predictor of disease progression (P =.0002). It is concluded that beyond a link to proliferation, high telomerase activity reflects a deregulation of the cell cycle associated with an increased rate of cells entering S phase and a higher degree of malignancy. Therefore, quantitative analysis of telomerase activity may be useful for identifying EC patients at high risk for recurrence. PMID- 11431716 TI - Atypical endometrial hyperplasia shares genomic abnormalities with endometrioid carcinoma by comparative genomic hybridization. AB - Endometrial hyperplasia is a common disorder that is now observed with increasing frequency in women treated with hormonal replacement therapy or with tamoxifen. This study was undertaken to determine whether genomic features of various forms of endometrial hyperplasias would allow their classification as a benign, premalignant, or malignant abnormality. Comparative genomic hybridization (CGH) was performed on endometrial glands microdissected by laser capture microscope from 19 archival endometrial samples, comprising 5 normal endometria, 1 polyp, 2 simple hyperplasias, 5 hyperplasias with nuclear abnormalities (atypical hyperplasias), and 4 low-grade and 2 high-grade endometrioid carcinomas, 1 with squamous component (adenoacanthoma). Genomic DNA, extracted from the glands and the squamous component in 1 case, was amplified by degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR) and compared with sex-matched DNA by CGH. No genomic imbalances were observed in the normal samples, the polyp, or the simple hyperplasias. However, in atypical hyperplasia, regardless of the level of cytologic atypia, genomic abnormalities were observed that also occurred in endometrioid carcinomas. Chromosomes 1, 8, and 10 were most often affected. The results are compared with molecular genetic abnormalities recently reported in these lesions. This study strongly suggests that atypical endometrial hyperplasias are closely related to endometrioid carcinoma and should be considered precancerous lesions, contrary to simple hyperplasia, which is a benign disorder. The squamous component of one of the high-grade carcinomas showed genetic abnormalities similar to those of endometrioid carcinoma and therefore does not represent squamous metaplasia but is an integral part of the malignant process. PMID- 11431717 TI - Acute funisitis of preterm but not term placentas is associated with severe fetal inflammatory response. AB - Acute funisitis, whose basic pathologic feature is umbilical vasculitis, constitutes a type of fetal inflammatory response to intrauterine infection. In the present study, a comparative analysis was performed between the clinicopathologic profiles of acute funisitis in term and preterm placentas along with measurement of fetal plasma interleukin 6 (IL-6) levels by specific immunoassay to assess the different biologic implications for the fetus. Acute funisitis in preterm placentas showed a significantly higher incidence of umbilical arteritis (P <.000001), higher fetal plasma IL-6 level (P <.0001), and higher prevalence of major perinatal morbidities (P <.0001). To assess the possible variation in fetal cell response to infectious agents according to gestational age, amnion cells and placental villous tissues obtained at different gestational ages were treated with bacterial lipopolysaccharides, and the IL-6 level of the culture media was assayed. Amnion cells and placental villous tissues from preterm placenta showed a more pronounced cytokine response than those from term placenta. The findings of this study indicate that the clinicopathologic significance of acute funisitis in term placentas is different from that of preterm placentas. Furthermore, they indicate that the robust inflammatory response of the fetus associated with elevated fetal plasma IL-6 level may reflect the biologic needs of the premature fetus to escape from the hostile intrauterine environment. PMID- 11431718 TI - Correlation of cystoscopic impression with histologic diagnosis of biopsy specimens of the bladder. AB - There is a paucity of information in the contemporary literature that would permit assessment of the urologist's ability to endoscopically discriminate between benign and malignant lesions of the bladder or to predict the grade and stage of papillary neoplasms. This prospective study evaluates the correlation between cystoscopic impression of urothelial lesions and final histologic diagnoses. Sixty-four patients with 68 urothelial abnormalities requiring formal biopsy or endoscopic resection were evaluated prospectively. At the time of endoscopy, treating urologists completed questionnaires documenting the surgeon's endoscopic impression of disease type and extent and performed standard biopsy or resection of all suspicious lesions. Specimens were submitted for routine histopathologic analysis, and the results were correlated with the questionnaire data. Endoscopic evaluation correctly discriminated between dysplastic/malignant and benign/reactive lesions in this study with a sensitivity of 100%, specificity of 100%, and positive and negative predictive values of 100%. Urologists could not readily distinguish between low- and high-grade papillary urothelial lesions and were frequently unable to determine if a tumor was invasive, particularly if the degree of invasion was microscopic. Endoscopic impression at the time of bladder biopsy or resection is accurate and discriminates between the presence and absence of cancer. Endoscopic impression alone is a relatively poor staging tool with respect to extent of invasive disease and must be coupled with careful histopathologic analysis of biopsy material, bimanual examination when appropriate, and axial imaging for complete assessment of a given tumor. PMID- 11431719 TI - Pancreatic intraepithelial neoplasia and infiltrating adenocarcinoma: analysis of progression and recurrence by DPC4 immunohistochemical labeling. AB - Pancreatic intraepithelial neoplasia (PanIN) is thought to be a precursor lesion of infiltrating pancreatic ductal adenocarcinoma (IPA). DPC4 is a tumor suppressor gene on chromosome 18q21.1 and is inactivated in approximately 55% of IPAs. Recently, immunohistochemical labeling using a monoclonal antibody to the Dpc4 protein has been shown to mirror DPC4 genetic status in invasive adenocarcinomas of the pancreas. In the present study, we examined the role of Dpc4 loss in neoplastic progression and recurrence. Two cases in which a PanIN clinically progressed to an invasive adenocarcinoma and a third of a patient with IPA of the head of the pancreas who later developed invasive adenocarcinoma in the tail of the pancreas were studied using Dpc4 immunolabeling. The first patient underwent pancreatic resection, which revealed PanIN-3 that lacked Dpc4 expression, and the patient developed an invasive pancreatic ductal carcinoma 10 years later that shared this loss of expression. The second patient had a pancreaticoduodenectomy for recurrent pancreatitis, and the resected pancreas contained PanIN-3 with intact Dpc4 expression. Seventeen months later, the patient developed an invasive adenocarcinoma of the distal pancreas that also had intact Dpc4 expression. In the third case, the patient underwent pancreaticoduodenectomy for an invasive ductal adenocarcinoma with negative margins. This carcinoma lacked Dpc4 expression. Three years later, resection of the pancreatic tail showed a second invasive adenocarcinoma. The cancer in the tail of the gland showed intact Dpc4 expression, suggesting it represented a second primary tumor, not a recurrence. We conclude that Dpc4 expression in PanIN can be predictive of Dpc4 expression in the subsequent invasive ductal adenocarcinoma. Additionally, Dpc4 expression can be used to differentiate recurrent or persistent adenocarcinoma from a second primary adenocarcinoma. PMID- 11431720 TI - Pathogenic significance of neuronal migration disorders in temporal lobe epilepsy. AB - To assess the epileptogenic lesions, a series of 202 cases with temporal lobectomy were analyzed histopathologically. The severity of hippocampal neuronal loss in patients with temporal lobe epilepsy was quantitatively analyzed and compared against autopsy controls of patients who died of nonneurologic disorders. For the histopathologic diagnosis of neuronal migration disorder (NMD), immunohistochemical stains for neurofilament protein (NF-M/H) and microtubule-associated protein 2 (MAP2) and Bielschowsky silver stains were routinely performed. Histopathology of NMD was classified by the 4-grade system. MAP2 immunoreactivity was useful in the identification of loss of normal polarization of dendrites in the abnormal neurons. NF-M/H immunohistochemistry and silver stains effectively labeled microscopic or occult lesions of NMD (grade II and III). Ammon hom sclerosis (AHS) was identified in 73.3% and NMD in 57.9%. There was more than 50% neuronal cell loss in 82.8% of AHS, and variable degrees of cell loss were observed in the dual-pathology groups. The frequency of dual pathology (both AHS and NMD) was 65.0% and showed relatively equal distributions in grades I, II, III, whereas the pure NMD group were classified predominantly as grades II and III. NMD might be a basic pathogenic substrate causing temporal lobe epilepsy. The dual pathology may indicate the presence of epileptogenic lesions in the neocortical and temporolimbic areas. PMID- 11431721 TI - Ultrastructural examination of the axillary skin biopsy in the diagnosis of metabolic diseases. AB - There is little information in the literature regarding the usefulness of ultrastructural examination of axillary skin biopsies in the evaluation of metabolic diseases. This is a retrospective clinicopathologic review of 143 patients who underwent axillary skin biopsies as part of evaluations for metabolic disease. Twenty-three (16%) had abnormalities, classified as follows: mitochondrial (n = 12), lysosomal (n = 6), increased glycogen (n = 3), nonspecific cytoplasmic inclusions (n = 2), ceroid lipofuscinosis (n = 1), and intradermal giant cells containing vacuoles and tubular inclusions (n = 1). Muscle biopsies were performed in 13 of the 23 patients; 11 showed abnormalities, including those related to mitochondria (n = 4) and other nonspecific changes (n = 7). Two patients underwent postmortem examination. Follow-up was available in 21 patients. A clinical or biochemical diagnosis was reached in 11 patients: metachromatic leukodystrophy (n = 2), electron transport chain abnormalities (n = 2), glutaric aciduria type II (n = 1), Unverricht disease (n = 1), Lennox-Gastaut syndrome (n = 1), ketotic hypoglycemia of childhood (n = 1), probable Leigh disease (n = 1), 5-methyl tetrahydrofolate homocystine methyltransferase deficiency (n = 1), and pyruvate dehydrogenase deficiency (n = 1). Of the 120 patients with negative skin biopsy results, 29 had abnormal findings on muscle (n = 27), nerve (n = 7), or brain (n = 3) biopsies. One patient had an abnormal heart biopsy result, and 3 patients underwent postmortem examinations. Follow-up was obtained in 27 of 29 patients. Diagnoses were achieved in 15 patients: electron transport chain abnormalities (n = 5), cortical dysplasia (n = 3), myoclonic epilepsy (n = 1), leukodystrophy (n = 2), Pallister-Killian mosaic syndrome (n = 1), Rett syndrome (n = 1), Landau-Kleffner syndrome (n = 1), and mitochondrial cardiomyopathy (n = 1). In conclusion, axillary skin biopsy is helpful in the evaluation of some causes of metabolic disease, but often the findings are nonspecific. A negative biopsy result does not rule out the possibility of metabolic disease, but a positive result may provide direction for further evaluation. PMID- 11431722 TI - Molecular characterization and sequence analysis of polyomavirus BKV-strain in a renal-allograft recipient. AB - The significance of polyomavirus (PV) infection was investigated in a 53-year-old patient who underwent renal transplantation and was treated with triple immunosuppressive therapy (tacrolimus, prednisone, and azathioprine). A renal biopsy taken because of the suspicion of acute rejection showed focal inflammatory interstitial infiltration, tubulitis, and tubular cell nuclear changes consistent with the hypothesis of viral infection. Both the tubular and decoy cells identified by means of urinalysis positively stained for anti-SV40 antibody. Polymerase chain reaction performed on the DNA extracted from renal tissue and isolated from urine showed the presence of an antigenic variant (AS) of the BKV archetype after sequence analysis of the transcription control region (TCR). On the basis of the diagnosis of BKV infection, immunosuppressive therapy was reduced. The patient's renal function improved and was still stable 8 months later when urinalysis showed only a few decoy cells, which were found to be infected by JC but not BK virus. These data suggest that only the BKV, probably favoured by immunosuppressive therapy (tacrolimus), causes renal damage. It is worth underlining that even small and sporadic viral genome mutations may lead to pathologic effects. PMID- 11431723 TI - Fibrillary glomerulonephritis: a report of 2 cases with extensive glomerular and tubular deposits. AB - Deposition of nonamyloid fibrillary material in glomeruli is well known. It is, however, unusual to find these fibrils in the tubular basement membranes and unprecedented to have fibrils of different sizes in the same patient. We present 2 cases with nephrotic range proteinuria with evidence of renal insufficiency. In both cases, strong, polyclonal immunoglobulin (Ig)G with C3 deposits were shown in the glomeruli and along tubular basement membranes. Ultrastructurally, the first case had 28-nm fibrils deposited extensively in the glomeruli and along tubular basement membranes. The second case had 30-nm fibrils in the glomeruli and 15-nm fibrils in the tubules. In both cases, the fibrils did not react with the regular amyloid stains. These findings are used to support the view that fibrillary glomerulopathy is not a disease, but rather the morphologic expression of an etiologically diverse group of diseases as yet incompletely defined. PMID- 11431726 TI - Is TRAIL hepatotoxic? PMID- 11431727 TI - Reduced mitochondrial adenosine triphosphate synthesis in skeletal muscle in patients with Child-Pugh class B and C cirrhosis. AB - Patients with cirrhosis of the liver often complain of tiredness and a lack of strength at physical exercise. Other investigators have found that muscle strength, work capacity, and maximal oxygen consumption are reduced in cirrhosis. We hypothesized that mitochondrial maximal rate of ATP synthesis in skeletal muscle may be impaired in these patients. This was tested with (31)P nuclear magnetic resonance spectroscopy in anterior tibial muscle of cirrhotic patients and healthy controls at rest, during exercise, and subsequent recovery. In patients with Child-Pugh class B and C cirrhosis resting PCr/P(i) ratio (8.3 +/- 1.0; n = 7) was lower than in patients with Child-Pugh class A cirrhosis (12.1 +/ 2.1; n = 7) and controls (11. 7 +/- 1.1; n = 6; P =.03), while the resting P(i)/gammaATP ratio was higher in Child-Pugh class B and C patients (0.43, 0.30, and 0.27, respectively; P =.03). Maximal rate of mitochondrial adenosine triphosphate (ATP) synthesis (V(max)) as calculated from the initial rate of phosphocreatine (PCr) recovery after work was lower in Child-Pugh class B and C cirrhosis (0.189 mmol/L/s +/- 0.034) than in both Child-Pugh class A patients (0.402 mmol/L/s +/- 0.103) and controls (0.425 mmol/L/s +/- 0.064; P =.01). V(max) was significantly correlated to intracellular free [Mg(2+)] obtained from the (31)P nuclear magnetic resonance (NMR) spectra (P =.003). Insufficient oxygen delivery did not seem a likely cause of reduced ATP synthesis in the patients. These findings suggest either a decreased number of mitochondria in skeletal muscle of the cirrhotic patient in Child-Pugh class B and C or a defective mitochondrial function that could be related to low intracellular free [Mg(2+)]. PMID- 11431728 TI - Oxidative stress in fatty livers of obese Zucker rats: rapid amelioration and improved tolerance to warm ischemia with tocopherol. AB - Fatty livers in humans and rats are less tolerant of ischemia, endotoxin, and alcohol. We hypothesized that fatty livers of obese (Ob) Zucker rats are oxidatively stressed and oxidative stress could be relieved by antioxidant treatment, leading to improved tolerance to ischemia. Total glutathione (GSH), tocopherol (TOC), ascorbic acid (AA), catalase (CAT), superoxide dismutase (SOD), and selenium-glutathione peroxidase (Se-GPx) were measured in the livers of Ob and lean (Ln) Zucker rats before and after treatment with high-dose TOC and ascorbate. Also, survival in treated Ob rats following a lethal 90 minutes of partial in vivo warm ischemia was examined. Fatty livers of Ob rats contained significantly less GSH, TOC, and CAT, in comparison with livers of Ln rats. Immunoblotting showed significantly decreased CAT protein without changes in mRNA in fatty livers. There were no significant differences in AA, SOD, and Se-GPx between the 2 groups. Pretreatment with TOC and ascorbate over 48 hours completely corrected the decreases in GSH, TOC, and CAT. Most importantly, TOC with or without ascorbate pretreatment significantly improved survival in Ob rats following ischemia in a dose-dependent manner. In conclusion, TOC administration corrected the oxidative stress in fatty livers of Ob Zucker rats and improved survival following lethal ischemia. Additional studies are needed to determine the efficacy of TOC-a relatively inexpensive agent-in treating patients with fatty livers in a variety of clinical conditions, possibly including liver transplantation. PMID- 11431729 TI - Hemodynamic, renal, and endocrine effects of acute inhibition of nitric oxide synthase in compensated cirrhosis. AB - To assess whether an increased production of nitric oxide is involved in the circulatory and renal alterations of cirrhosis, we evaluated systemic hemodynamics (echocardiography), renal hemodynamics, and sodium handling (lithium clearance method), plasma renin activity (PRA), aldosterone (PAC), and norepinephrine in 7 patients (3 men, mean age 65 +/- 2 years) with compensated cirrhosis, portal hypertension, and hyperdynamic circulation during intravenous N(G)-monomethyl-L-arginine (L-NMMA) (3 mg/kg bolus plus 0.05 mg/kg. min for 120 minutes) or placebo (the vehicle) in a randomized, placebo-controlled, crossover study. Administration of L-NMMA resulted in significant reductions in plasma and urinary nitrite levels and plasma cyclic guanosine monophosphate (cGMP), indicating effective inhibition of nitric oxide synthase. L-NMMA also significantly reduced cardiac index (-13%) and increased systemic vascular resistance (+26%), arterial pressure (+9%), renal blood flow (+12%), glomerular filtration rate (+12%), and sodium excretion (+25%). Changes in sodium excretion were caused by both enhanced filtered sodium load and reduced sodium reabsorption in the proximal tubule. Plasma norepinephrine significantly decreased in response to L-NMMA, and there was a trend for reductions in PRA and PAC. Placebo had no appreciable effect on any of the measured parameters. These results indicate that in patients with compensated cirrhosis, portal hypertension and hyperdynamic circulation inhibition of nitric oxide synthase corrects the altered systemic hemodynamics and improves renal function and sodium excretion. PMID- 11431730 TI - Immunogenicity of hepatitis A vaccination in decompensated cirrhotic patients. AB - Hepatitis A virus (HAV) vaccination is recommended in chronic liver disease because of an increased morbidity and mortality associated with HAV superinfection. However, data regarding the efficacy of HAV vaccination in patients with advanced chronic liver disease is limited. We assessed the efficacy of a standard HAV vaccination schedule in decompensated chronic liver disease in comparison with compensated disease and defined clinical predictors associated with seroconversion. Eighty-four anti-HAV antibody-negative patients, 49 with compensated liver disease, and 35 with decompensated disease were enrolled. Seroconversion was measured by qualitative and quantitative anti-HAV antibody measurements 1 month after each vaccine dose, and univariate/multivariate analysis was performed to define clinical predictors associated with seroconversion. One month after the primary dose, 71.4% of patients with compensated liver disease had detectable anti-HAV antibody compared with 37.1% with decompensated liver disease (P <.05). One month after the booster dose, 98% of compensated patients seroconverted compared with 65.7% with decompensated disease (P <.05). The median serum antibody concentration in compensated liver disease was 76.4 mIU/mL at month 1 and 327.91 mIU/mL at month 7 compared with 20.0 mIU/mL and 102.57 mIU/mL, respectively, in decompensated disease. On multivariate analysis, Child-Pugh class was the only factor predicting response to vaccination. Seroconversion after HAV vaccination was significantly less common in decompensated liver disease and the presence of advanced disease (Child Pugh class B/C) predicted a lower response rate. These findings indicate that the response to HAV vaccination in chronic liver disease is optimal when targeted to patients before the development of hepatic decompensation. PMID- 11431731 TI - Roles of Akt/PKB and IKK complex in constitutive induction of NF-kappaB in hepatocellular carcinomas of transforming growth factor alpha/c-myc transgenic mice. AB - NF-kappaB regulates liver cell death during development, regeneration, and neoplastic transformation. For example, we showed that oncogenic Ras- or Raf mediated transformation of rat liver epithelial cells (RLEs) led to altered NF kappaB regulation through IKK complex activation, which rendered these cells more resistant to TGF-beta1-induced apoptosis. Thus, based on these findings, we sought to determine whether NF-kappaB could also be involved in tumor growth of liver cells in vivo. Hepatocellular carcinomas (HCCs) derived from bitransgenic mice harboring TGF-alpha and c-myc transgenes targeted specifically to the liver were compared with HCCs from c-myc single transgenic mice. Tumors from bitransgenic mice are characterized by a higher frequency of appearance, lower apoptotic index, and a higher rate of cell proliferation. Here we show that NF kappaB is activated in HCCs of double TGF-alpha/c-myc transgenic mice, but not of c-myc single transgenic mice, suggesting that TGF-alpha mediates induction of NF kappaB. Activation of the IKK complex was observed in the HCCs of double TGF alpha/c-myc transgenic mice, implicating this pathway in NF-kappaB induction. Lastly, activation of the Akt/protein kinase B (PKB), which has recently been implicated in NF-kappaB activation by PDGF, TNF-alpha, and Ras, was also observed. Importantly, human HCC cell lines similarly displayed NF-kappaB activation. Thus, these studies elucidate an anti-apoptotic mechanism by a TGF alpha-Akt/PKB-IKK pathway, which likely contributes to survival and proliferation, thereby accelerating c-myc-induced liver neoplastic development in vivo. PMID- 11431732 TI - The integrin, alpha6beta1, is necessary for the matrix-dependent activation of FAK and MAP kinase and the migration of human hepatocarcinoma cells. AB - Expression of the integrin, alpha6beta1, a receptor for laminins, is associated with the progression of hepatocellular carcinoma (HCC). The approach to investigating the alpha6beta1 integrin signaling in HCC cells was to express a deletion mutant of the beta4 integrin cytoplasmic domain (beta4-Deltacyt) in 2 HCC cell lines, HepG2 and Huh7. Expression of this mutant prevents formation of the alpha6beta1 heterodimer. As expected, adhesion of both the HepG2/beta4 Deltacyt and Huh7/beta4-Deltacyt transfectants to laminin, but not to collagen, was reduced compared with the mock transfectants. However, migration of the beta4 Deltacyt transfectants toward both collagen and laminin was inhibited, suggesting a role for alpha6beta1 in the signaling of migration. Migration of HCC cells requires mitogen-activated protein (MAP) kinase. The adhesion of the beta4 Deltacyt transfectants to collagen resulted in a substantial reduction in MAP kinase activation in comparison with the mock transfectants, although their ability to activate MAP kinase in response to epidermal growth factor (EGF) stimulation was not impaired. In addition, matrix adhesion of the beta4-Deltacyt transfectants did not stimulate the tyrosine phosphorylation of focal adhesion kinase (FAK), and this defect correlated with reduced binding of adaptor protein Grb2 to FAK. These results suggest that FAK tyrosine phosphorylation is dependent on alpha6beta1 expression, and that FAK-Grb2 association plays a central role in alpha6beta1-mediated activation of MAP kinase. Moreover, the expression of alpha6beta1 in HCC cells is necessary for FAK/MAP kinase-dependent migration. PMID- 11431733 TI - Restoration of cerebral blood flow autoregulation and reactivity to carbon dioxide in acute liver failure by moderate hypothermia. AB - In patients with acute liver failure (ALF) and uncontrolled intracranial hypertension, moderate hypothermia (32 degrees C) reduces intracranial pressure (ICP) and cerebral blood flow (CBF), and can be used as a bridge to liver transplantation. The purpose of this study was to test the hypothesis that moderate hypothermia reduced ICP by restoring CBF autoregulation. Nine patients with uncontrolled intracranial hypertension and ALF who fulfilled the criteria for poor prognosis were studied. CBF autoregulation and reactivity to carbon dioxide were evaluated before and 4 hours after cooling (32 degrees C). Significant reductions were observed in the ICP (median, 46 [range, 27-54] mm Hg to 19 [15-22] mm Hg; P <.01) and CBF (median, 111 [69-134] to 56 [38-67] mL/100 g/min; P <.05). The defective CBF autoregulation and the absence of reactivity to carbon dioxide that was observed in all patients was restored with cooling. The results of our study suggest that the improvement in ICP observed with hypothermia may be the result of its effects on CBF autoregulation and provides a tool to explore the mechanisms associated with the deranged CBF autoregulation in ALF. PMID- 11431734 TI - Expression and role of Bcl-xL in human hepatocellular carcinomas. AB - Transformed hepatocytes survive various apoptotic insults during their growth in vivo. However, molecular mechanisms that inhibit apoptosis and support their survival are not well understood. In this study, we investigated the expression and role of Bcl-xL, an antiapoptotic member of the Bcl-2 family, in human hepatocellular carcinoma (HCC). The Bcl-xL protein was expressed in HepG2, Hep3B, and Huh7 human hepatoma cell lines at high levels, but none of these cells expressed Bcl-2. Down-modulation of Bcl-xL by antisense oligonucleotide activated apoptosis in HepG2 cells in response to cellular stresses induced by staurosporine treatment or by serum starvation. Ectopic expression of transcriptionally active p53 alone was not sufficient for the activation of apoptosis in p53-null Hep3B cells, but apoptosis was induced when endogenous Bcl xL was simultaneously inhibited by antisense oligonucleotide in these cells. Bcl xL was expressed in all 20 surgically resected human HCC tissues when examined by Western blot analysis and immunohistochemistry, and levels of its expression were higher in a subset of HCC tissues than those of adjacent nontumor liver tissues or normal livers. We conclude that Bcl-xL expressed in human HCC cells inhibits apoptosis produced by various cellular stresses, such as staurosporine treatment, serum starvation, and p53 activation, and may play an important role in their survival. PMID- 11431735 TI - Autocrine motility factor enhances hepatoma cell invasion across the basement membrane through activation of beta1 integrins. AB - Autocrine motility factor/phosphohexose isomerase (AMF/PHI) is a cytokine that is linked to tumor invasion and metastasis. In hepatocellular carcinoma (HCC) tissues, hepatoma cells produce AMF/PHI and its receptor, Mr 78,000 glycoprotein (gp78), is strongly detected in hepatoma cells invading into the stroma and tumor thrombi in the portal vein. Here, we investigated the mechanism of hepatoma cell invasion through Matrigel induced by AMF/PHI using 3 hepatoma cell lines. Production of AMF/PHI, phosphorylation of MEK1/2, and Rho activity were investigated by immunoblotting. Expression of AMF/PHI and gp78 was observed by confocal fluorescence microscopy. The influence of AMF/PHI on activated integrin beta1 subunit expression was evaluated by flow cytometry. Changes in invasion, adhesion, and motility induced by AMF/PHI were evaluated using chemoinvasion, adhesion, and phagokinetic track motility assays. The effect of AMF/PHI on matrix metalloproteinase (MMP) secretion was evaluated by gelatin zymography. Hepatoma cells produced AMF/PHI and expressed gp78. Although AMF/PHI was ubiquitously detected, gp78 was strongly expressed in migrating cells. AMF/PHI induced up regulation of activated integrin beta1 subunit expression. AMF/PHI stimulated hepatoma cell invasion through Matrigel, and stimulated the adhesion, motility, and MMP-2 secretion of hepatoma cells. The latter effects were suppressed by the function-blocking antibody for integrin beta1 subunit. AMF/PHI also enhanced Rho activity and the phosphorylation of MEK1 and MEK 2. Our results indicate that AMF/PHI enhances hepatoma cell invasion through Matrigel in an autocrine manner by stimulating the adhesion, motility, and MMP-2 secretion of these cells through activation of beta1 integrins. PMID- 11431736 TI - Adenovirus-mediated CD40 ligand gene therapy in a rat model of orthotopic hepatocellular carcinoma. AB - Unresectable hepatocellular carcinoma (HCC) lacks effective therapy and entails very poor prognosis. In this study, we have explored a gene-therapeutic approach to stimulate antitumor immunity by adenoviral-mediated transfer of CD40 ligand to treat HCC in rats. In vitro infection of a rat HCC cell line (McA-RH7777) with adenoviral vector expressing CD40 ligand (AdCMVmCD40L) induced CD40L expression in a dose-dependent manner. Expression of CD40L in McA-RH7777 cells did not alter their growth rate in vitro, but it abrogated their tumorigenicity when CD40L expressing cells were implanted into the liver of syngenic Buffalo rats. In vivo gene therapy of established orthotopic HCC nodules (6.5 mm in diameter) in Buffalo rats by intratumor injection of AdCMVmCD40L vector led to complete tumor eradication and long-term survival in 69.5% of treated animals. Therapy with AdCMVmCD40L induced strong lymphocytic infiltration of the tumoral tissue and increased apoptosis of malignant cells. The observed antitumoral effect was mediated by CD8(+) T cells and was associated with increased interleukin (IL)-12 serum levels and enhanced natural killer (NK) activity. Animals that eliminated the tumor after in vivo gene therapy developed protective antitumor immunity being resistant to rechallenge with neoplastic cells. Toxicity of the therapy with AdCMVmCD40L was slight, with only a transient increase in the level of serum transaminases and minor lymphocyte infiltration of normal liver tissue. These data demonstrate that intratumoral administration of AdCMVmCD40L may provide an efficient and safe treatment for HCC. PMID- 11431737 TI - Increased extracellular matrix remodeling is associated with tumor progression in human hepatocellular carcinomas. AB - Matrix metalloproteinase-2 (MMP2) is a key enzyme in the process of extracellular matrix remodeling involved in tumor invasion and metastasis. The activation of MMP2 involves interplay with the membrane type-matrix metalloproteinase-1 (MT1 MMP) and the tissue inhibitor of metalloproteinase-2 (TIMP2). In vitro, activated hepatic stellate cells are a main source of MMP2 and collagen I induces MMP2 activation. The steady-state mRNA levels of MMP2, MT1-MMP, TIMP2, collagen I, collagen IV, and laminin gamma1 were compared with MMP2 activity in 55 hepatocellular carcinomas, 47 matching nontumor biopsies and 19 histologically normal livers. In hepatocellular carcinomas, increased collagen I mRNA levels were strongly associated with those of MMP2 (Spearman R =.74, P <.001), MT1-MMP (R =.65, P <.001) and TIMP2 (R = 0.61, P <.001). MMP2 activity was correlated with the mRNA expression of collagen I (R =.45 P <.01), collagen IV (R =.40, P <.01) and laminin gamma1 (R =.33, P <.05). Unlike collagen IV and laminin gamma1 mRNAs, MMP2, MT1-MMP, TIMP2, collagen I mRNA levels were increased in nonencapsulated compared with encapsulated tumors (P <.05). In addition, MMP2 activity was fourfold higher (P <.01) in tumors arising in cirrhotic livers than in those arising in noncirrhotic livers. Moreover, tumor recurrence was associated with 4.6- and 2.8-fold (P <.05) higher collagen I and MMP2 mRNA levels, respectively, in hepatocellular carcinomas arising in cirrhotic livers. Thus, a high extracellular matrix remodeling favors tumor progression in hepatocellular carcinomas. PMID- 11431738 TI - The role of Smad3 in mediating mouse hepatic stellate cell activation. AB - Transforming growth factor beta (TGF-beta) is the most potent profibrogenic mediator in liver fibrosis. Although Smad proteins have been identified as intracellular mediators in the TGF-beta signaling pathway, the function of individual Smad proteins remains poorly understood. The aim of this study was to explore the contribution of Smad3 in mediating TGF-beta responses in a model of acute liver injury in vivo and in culture-activated hepatic stellate cells (HSCs). Wild-type, Smad3 heterozygous or Smad3 homozygous knockout mice were treated with a single intragastric administration of CCl(4). After 72 hours, the induction of hepatic collagen alpha1(I) and alpha2(I) messenger RNA (mRNA) levels in Smad3 knockout mice was only 42% and 64%, respectively, of the levels induced in wild-type mice. However, smooth muscle alpha-actin (alpha-SMA) was expressed at a slightly higher level in livers from knockout mice compared with wild-type mice. In culture-activated HSCs from Smad3 knockout mice, collagen alpha1(I) mRNA was 73% of wild-type HSCs, but alpha-SMA expression was the same. HSCs from knockout mice showed a higher proliferation rate than wild-type HSCs. Smad3 deficient HSCs did not form TGF-beta1-induced Smad-containing DNA-binding complexes. In conclusion, (1) maximal expression of collagen type I in activated HSCs requires Smad3 in vivo and in culture; (2) Smad3 is not necessary for HSC activation as assessed by alpha-SMA expression; (3) Smad3 is necessary for inhibition of proliferation of HSCs, which might be TGF-beta-dependent; and (4) Smad3 is required for TGF-beta1-mediated Smad-containing DNA-binding complex formation in cultured HSCs. PMID- 11431739 TI - Toll-like receptor 4 is involved in the mechanism of early alcohol-induced liver injury in mice. AB - Chronic alcohol administration increases gut-derived endotoxin in the portal blood, which activates Kupffer cells and causes liver injury. Mice (C3H/HeJ) with mutations in toll-like receptor 4 (TLR4) are hyporesponsive to endotoxin. To test the hypothesis that TLR4 is involved in early alcohol-induced liver injury, the long-term intragastric ethanol feeding protocol developed by Tsukamoto and French for rats was adapted to mice. Animals with nonfunctional TLR4 and wild-type mice (C3H/HeOuJ) were compared. Two-month-old female mice were fed a high-fat liquid diet with either ethanol or isocaloric maltose-dextrin as control continuously for 4 weeks. There was no difference in mean urine alcohol concentrations between the groups. Dietary alcohol significantly increased liver-to-body weight ratios and serum alanine transaminase (ALT) levels in wild-type mice (109 +/- 18 U/L) over high-fat controls (40 +/- 3 U/L), effects that were blunted significantly in mice with a mutation of TLR4 (55 +/- 9 U/L). While no significant pathologic changes were observed in high-fat controls, dietary ethanol caused steatosis, mild inflammation, and focal necrosis in wild-type animals (pathology score = 5.2 +/- 1.2). These pathologic changes were significantly lower in TLR4-deficient mice fed ethanol (score = 2.0 +/- 1.3). Endotoxin levels in the portal vein were increased significantly after 4 weeks in both groups fed ethanol. Moreover, ethanol increased tumor necrosis factor alpha (TNF-alpha) mRNA expression in wild type, but not in TLR4-deficient, mice. These data are consistent with the hypothesis that Kupffer cell activation by endotoxin via TLR4 is involved in early alcohol-induced liver injury. PMID- 11431740 TI - Temporal changes in insulin sensitivity following the development of acute liver failure secondary to acetaminophen. AB - Insulin resistance in chronic liver disease (CLD) is well documented. This study investigated whether similar changes occur in acute liver failure (ALF). Patients with ALF (n = 10) were recruited within 72 hours of their peak prothrombin time (range 42-120 seconds). All patients were ventilated for encephalopathy (grade III-IV). Peripheral and endogenous insulin sensitivity were assessed by a hyperinsulinemic euglycemic clamp (Human Actrapid 1.5 mU/min/kg) with an infusion of D-[6,6-(2)H(2)] glucose. The clamp was performed on day 0 and then on day 7 and day 14. During the insulin infusion, the mean total peripheral glucose uptake (area under the curve [AUC]) was 1,422 (SD, 1,253), 2,244 (SD, 1,392), and 4,500 (SD, 1,120) micromol/kg on days 0, 7, and 14, respectively. Significant changes occurred from day 0 to 14 (day 14-day 0: 3,078 [95% CI, 1,798 to 4,359]; P =.001) and day 7 to 14 (day 14-day 7: 2,256 [95% CI, 923 to 3,589]; P =.001). No significant difference in endogenous glucose production was demonstrated over time. Mean peripheral insulin sensitivity altered over time, increasing from 0.09 (SD, 0.09) micromol/kg/min/mU/L on day 0 to 0.24 (SD, 0.16) on day 7 and 0.5 (SD, 0.1) on day 14. Significant changes occurred between days 0, 7, and 14 (day 7-day 0: 0.15 [95% CI, 0.04 to 0.26], P =.006; day 14-day 0: 0.4 [95% CI, 0.28 to 0.5], P =.001; day 14-day 7: 0.2 [95% CI, 0.12 to 0.38], P =.001). This study demonstrates that in ALF, impaired peripheral uptake of glucose occurs, peripheral insulin sensitivity being restored at 2 weeks in subjects who survived. PMID- 11431741 TI - Navajo neurohepatopathy: a mitochondrial DNA depletion syndrome? AB - Navajo neurohepatopathy (NNH) is an autosomal recessive disease of full-blooded Navajo children living in the Navajo Reservation of southwestern United States. Clinical features of NNH include peripheral and central nervous system involvement, acral mutilation, corneal scarring or ulceration, liver failure, and metabolic and immunologic derangement. The cause of NNH is unknown, but the clinical features of NNH are similar to those of patients with mitochondrial DNA (mtDNA) depletion. Therefore, we studied mtDNA concentration in the liver from 2 patients with NNH. Using histochemical, biochemical, and molecular techniques, we found evidence of mtDNA depletion, and we propose that the primary defect in NNH is in the nuclear regulation of mtDNA copy number. PMID- 11431742 TI - Cigarette smoking and hepatic lesions in patients with chronic hepatitis C. AB - A possible hepatotoxicity of cigarette smoke has been recently suggested by epidemiological and experimental studies. Our aim was to study the possible relationships between smoking and liver fibrosis and activity in patients with chronic hepatitis C. A cross-sectional study was performed in a group of 310 patients with chronic hepatitis C consecutively hospitalized for their first liver biopsy. The relationships between age, gender, alcohol consumption, route of contamination, tobacco consumption, and Knodell fibrosis and activity scores were examined in univariate, age-adjusted, and multivariate analyses. One hundred and seventy-six patients (57%) were current smokers. Smokers were younger (P <.001), more often of male gender (P =.001), more often alcohol consumers (P =.001), and more often had a history of intravenous drug use (P =.0001) than never smokers. Smoking was related to increased fibrosis and activity scores in age-adjusted (P =.009 and P =.005, respectively) and multivariate analyses (P =.03 and P =.04, respectively). Smoking increases the severity of hepatic lesions in patients with chronic hepatitis C. PMID- 11431743 TI - Liver transplantation in patients with hepatitis B virus infection: outcome in Asian versus white patients. AB - Previous studies have found that Asian patients transplanted for hepatitis B virus (HBV) infection had worse outcomes than white patients. The aim of this study was to compare outcomes in Asian and white patients listed for liver transplantation for HBV infection. Data of all patients with HBV infection listed for liver transplantation between January 1996 and June 1998 from 20 centers in North America were collected using a survey. Total patients enrolled were 325 (171 whites, 126 Asians, 28 other races). There was no difference in demographics, liver biochemistry, and HBV replicative status between Asians and whites at the time of listing. More Asians had hepatocellular carcinoma and fewer Asians had hepatitis C or D virus coinfection. At the time of this survey, 70 Asians (55%) and 99 whites (58%) had been transplanted. Actuarial 2-year survival posttransplantation for Asians (88%) and whites (92%) was similar. Recurrent HBV infection occurred in 8 (11%) Asians and 12 (12%) whites. Five patients with recurrent HBV infection died, 4 of whom were Asian. Actuarial 2-year survival for Asians versus whites with recurrent HBV infection was 60% versus 90% (P =.04). In this large cohort of patients, overall survival and recurrent HBV infection posttransplantation were comparable between Asians and whites. However, Asians with recurrent HBV infection posttransplantation had significantly higher mortality. PMID- 11431744 TI - A randomized 4-arm multicenter study of interferon alfa-2b plus ribavirin in the treatment of patients with chronic hepatitis C not responding to interferon alone. AB - To determine whether a higher dosage of interferon (IFN) associated with ribavirin and/or prolonged time of administration may improve therapeutic efficacy, we conducted a 4-arm randomized trial on patients with chronic hepatitis C not responding to one or more previous treatment courses with IFN monotherapy. Group 1 (n = 139) received 3 million units (MU) IFN-alpha2b 3 times a week (t.i.w.) plus ribavirin 1,000 mg/d for 12 months; group 2 (n = 162) received 5 MU t.i.w. plus ribavirin for 12 months; group 3 (n = 142) received 3 MU t.i.w. plus ribavirin for 6 months; and group 4 (n = 151) received 5 MU t.i.w. plus ribavirin for 6 months. The primary end point was hepatitis C virus (HCV) RNA clearance at the end of 6-month follow-up. HCV-RNA was negative in 15% of group 1, 23% of group 2, 11% of group 3, 16% of group 4 (group 2 vs. group 3, P =.04). Among patients with genotypes 1 and 4, sustained response was significantly higher in group 2 vs. group 3 (18% vs. 7%, P =.03; group 1 = 9%, group 4 = 12%, P = not significant [NS]). In patients with genotypes 2 and 3, sustained virologic response was not affected by the different regimens (group 1 = 32%, group 2 = 30%, group 3 = 30%, group 4 = 35%, P = NS). In conclusion, about 23% of nonresponders to IFN monotherapy may achieve a sustained response if re treated by 5 MU t.i.w. IFN plus ribavirin 1,000 mg/d for 1 year. Patients with genotype 1 should receive a high dosage of IFN plus ribavirin for 12 months, whereas therapy for patients with genotype 2 or 3 should be less aggressive. PMID- 11431745 TI - Long-term follow-up of interferon alfa treatment in Chinese patients with chronic hepatitis B infection: The effect on hepatitis B e antigen seroconversion and the development of cirrhosis-related complications. AB - The long-term effect of interferon alfa (IFN-alpha) in Chinese patients with chronic hepatitis B infection is unknown. A total of 411 chronic hepatitis B patients (208 treated with IFN-alpha and 203 as control) were followed up for hepatitis B serology and the development of hepatoma and other cirrhosis-related complications. The hepatitis B e antigen (HBeAg) seroconversion rate in the IFN alpha-treated group, though significantly greater at 6 and 24 months, was comparable with the control group on subsequent follow-up, irrespective of pretreatment alanine transaminase (ALT) levels. HBeAg seroreversion rate was higher in the IFN-alpha group compared with the control group (21.1% vs. 2.2%; P =.001). Loss of hepatitis B surface antigen (HBsAg) occurred in 2.4% of the IFN alpha-treated patients and 0.49% of the control patients (P = NS). Around 90% of the anti-HBe-positive patients in both groups were still hepatitis B virus (HBV) DNA-positive by polymerase chain reaction (PCR) assay. Two patients suffered from hepatic reactivation during the course of treatment. Nine (4.3%) patients in the IFN-alpha group and 2 (1.0%) in the control group developed complications of cirrhosis and hepatoma (P =.062). In Chinese HBsAg carriers, IFN-alpha was of no long-term benefit in inducing HBeAg seroconversion or in the prevention of hepatoma and other cirrhosis-related complications. PMID- 11431746 TI - A cellular gene up-regulated by hepatitis B virus-encoded X antigen promotes hepatocellular growth and survival. AB - Polymerase chain reaction (PCR) select complementary DNA (cDNA) subtraction of hepatitis B x antigen (HBxAg)-positive compared with -negative HepG2 cells resulted in the up-regulated expression of a cellular gene that encodes a transcript of 745 bases and a polypeptide 99 amino acids long. GenBank analysis revealed extensive homology with the amino terminal domain of cellular multidrug resistant proteins (MRP), although overexpression of this gene did not confer an MRP phenotype. In situ hybridization and immunostaining showed colocalized expression with HBxAg in the liver of hepatitis B carriers. Overexpression of this protein stimulated the growth of HepG2 cells in serum-free medium, and partially protected cells from anti-Fas-mediated killing, but did not promote growth in soft agar or tumor formation in nude mice. Introduction of the dominant negative inhibitor of nuclear factor kappaB (IkappaBalpha) into HBxAg-positive HepG2 cells decreased the levels of messenger RNA (mRNA) and protein, suggesting that its up-regulation is nuclear factor kappaB (NF-kappaB) dependent. Hence, HBxAg activation of NF-kappaB may result in the up-regulation of a cellular protein that promotes growth factor-independent survival and protects against Fas mediated killing. This factor may contribute to the persistence of infected hepatocytes during chronic infection, which is important for the later development of hepatocellular carcinoma (HCC). PMID- 11431747 TI - High amino acid variability within the NS5A of hepatitis C virus (HCV) is associated with hepatocellular carcinoma in patients with HCV-1b-related cirrhosis. AB - Interferon therapy may decrease the risk of hepatocellular carcinoma in patients with hepatitis C virus (HCV)-related liver cirrhosis. Interaction of the cellular protein kinase PKR with the PKR-binding domain (PKR-bd) of HCV-NS5A protein may affect cellular growth control and viral resistance to interferon therapy. Mutations within the PKR-bd, which comprises the interferon sensitivity determining region (ISDR), have been associated with interferon sensitivity. To determine whether or not there is an association between HCV heterogeneity and the presence of hepatocellular carcinoma, HCV-1b genomic regions were amplified and directly sequenced from serum samples obtained from 82 patients with liver cirrhosis, 53 with, and 29 without hepatocellular carcinoma. None of them had received antiviral therapy. When compared with the deduced consensus sequence, the median number of amino acid changes in the PKR-bd was higher among samples from patients with (4.22) than from those without hepatocellular carcinoma (1.62; P <.001), and isolates with 3 or more amino acid changes were significantly more common among the former (60%) than among the later (6%, P <.001). No such differences were observed in other viral regions, including Core, E2-HVR-1, E2 PePHD, NS3, and the 5' and 3' PKR-bd flanking regions. In addition, amino acid variation in viral regions other than HVR-1 did not accumulate over time in the analyzed sequential serum samples obtained from patients with or without hepatocellular carcinoma. Therefore, a mutated HCV-PKR-bd phenotype is very common in cirrhotic patients with hepatocellular carcinoma. PMID- 11431748 TI - Initial amplification of duck hepatitis B virus covalently closed circular DNA after in vitro infection of embryonic duck hepatocytes is increased by cell cycle progression. AB - The relationship between the cell cycle and early amplification of duck hepatitis B virus covalently closed circular (CCC) DNA was studied after in vitro infection of fetal hepatocytes. We first showed that embryonic hepatocytes proliferated for at least 6 days after plating and that complete viral replication including CCC DNA amplification occurred in these proliferating cells. Addition of sodium butyrate or aphidicolin reversibly blocked cells in the G1 phase and diminished CCC DNA synthesis, which was restored after drug withdrawal, concomitantly with the entry of cells into S phase. Cell cycle progression of fetal hepatocytes can be triggered by stimulation with epidermal growth factor (EGF), hepatocyte growth factor (HGF), and tumor growth factor alpha (TGF-alpha). CCC DNA synthesis increased with progression to the S phase induced by EGF, HGF, and TGF-alpha alone or in combination. By contrast, tumor growth factor beta (TGF-beta) alone or in combination with EGF inhibited cell proliferation and viral DNA synthesis. By double labeling, viral nucleocapsids were found predominantly in bromodeoxyuridine-positive hepatocytes, indicating that high viral replication occurs preferentially in proliferating hepatocytes. CCC DNA was also detected mainly in cells in the S and G2/M phases separated from cells in the G1 phase by cell sorting. Taken together, these results show that hepatocyte proliferation may positively regulate the initial amplification of CCC DNA of avian hepadnaviruses, and may explain why mitosis is not necessarily associated with loss of CCC DNA. PMID- 11431749 TI - Hepatitis C virus infection and needle exchange use among young injection drug users in San Francisco. AB - Young injection drug users (IDUs) in San Francisco may be at high risk for hepatitis C virus (HCV) infection despite access to several needle exchange venues. The authors conducted a cross-sectional study from 1997 to 1999 in San Francisco to estimate the prevalence and incidence of antibody to HCV (anti-HCV) among street-recruited IDUs under age 30, and to examine risk behaviors and sources of sterile needles. Among 308 participants, the prevalence of anti-HCV was 45%. Using statistical modeling, incidence of HCV infection was estimated to be 11 per 100 person years. Independent risk factors for anti-HCV included age (odds ratio [OR], 1.17 per year; 95% confidence interval [CI], 1.05-1.30), years injecting (OR, 1.21 per year; 95% CI, 1.10-1.34), years in San Francisco (OR, 1.06 per year; 95% CI, 1.00-1.14), first injected by a sex partner (OR, 4.06; 95% CI, 1.74-9.52), injected daily (OR, 3.85; 95% CI, 2.07-7.17), ever borrowed a needle (OR, 2.56; 95% CI, 1.18-5.53), bleached last time a needle was borrowed (OR, 0.50; 95% CI, 0.24-1.02), snorted or smoked drugs in the prior year (OR, 0.48; 95% CI, 0.26-0.89), and injected by someone else in the prior month (OR, 0.50; 95% CI, 0.25-0.99). In the prior month, 88% used at least 1 of several needle exchange venues, and 32% borrowed a needle. We conclude that anti-HCV prevalence is lower than in previous studies of older IDUs, but 11% incidence implies high risk of HCV infection in a long injecting career. Despite access to sterile needles, borrowing of needles persisted. PMID- 11431750 TI - Treatment of hepatitis C infection in injection drug users. AB - Chronic hepatitis C is the most common infectious disease among injection drug users (IDUs). Because of the allegedly poor compliance of IDUs with treatment requirements and conditions, hepatologists recommend treatment only if former IDUs have spent 6 to 12 months drug free. The aim of this prospective study was to investigate whether opiate-dependent IDUs with chronic hepatitis C virus (HCV) infection can be treated successfully with interferon. Eligibility for the study meant IDUs had to be HCV-RNA positive by polymerase chain reaction. Subsequently 50 inpatients were enrolled during detoxification treatment. HCV treatment was started with interferon alfa-2a (through 1998) or a combined regimen consisting of interferon alfa-2a and ribavirin (begun in 1998). All patients were treated and supervised by specialized physicians in both hepatology and addiction medicine. The end point for this study was defined as a loss of detectable serum HCV RNA at week 24 after treatment. The rate of sustained virologic response was 36%. Sustained response rates were not significantly different for patients who relapsed and returned to treatment (53%), relapsed and did not return to treatment (24%), or did not relapse (40%; P >.05). During the 24 weeks after treatment, we were unable to detect any reinfection, even among patients who injected heroin during this period. This surprising result should be examined in further studies. In conclusion, HCV-infected drug addicts with chronic HCV infection can be treated successfully with interferon alfa-2a and ribavirin if they are closely supervised by physicians specialized in both hepatology and addiction medicine. PMID- 11431751 TI - Persistent hepatitis B virus infection in subjects without hepatitis B surface antigen: clinically significant or purely "occult"? PMID- 11431752 TI - Occult hepatitis B virus infection: a hidden menace? PMID- 11431753 TI - Gene therapy for hepatocellular carcinoma-teaching old dogs new tricks. PMID- 11431754 TI - Alcohol-induced myopathy: what is the role of mitochondria? PMID- 11431755 TI - Bloody liver-prometheus revised. PMID- 11431756 TI - To MELD or not to MELD? PMID- 11431759 TI - Emergency department ultrasonography in the evaluation of hypotensive and normotensive children with blunt abdominal trauma. AB - PURPOSE: The aim of this study was to evaluate the accuracy of emergency department (ED) ultrasound scan in identifying which children with blunt torso trauma have intraperitoneal fluid associated with intraabdominal injuries (IAI). METHODS: The authors conducted a prospective, observational study of children (< 16 years old) with blunt trauma who presented to a level 1 trauma center over a 29-month period and underwent abdominal ultrasound scan while in the ED. Ultrasound examinations were ordered at the discretion of the trauma surgeons or ED physicians caring for the patients, performed by trained sonographers, and interpreted at the time of the ultrasound. Ultrasound examinations were interpreted solely for the presence or absence of intraperitoneal fluid. Hypotension was defined as > or = 1 standard deviation below the age-adjusted mean. Patients underwent follow-up to identify those with intraperitoneal fluid and IAI. RESULTS: A total of 224 pediatric blunt trauma patients had ultrasound scan performed and were enrolled. Thirty-three patients had IAI with intraperitoneal fluid, and ultrasound scan was positive in 27. The accuracy of abdominal ultrasound for detecting intraperitoneal fluid associated with IAI was sensitivity, 82% (95% confidence interval [CI] 65% to 93%); specificity, 95% (95% CI 91% to 97%); positive predictive value, 73% (95% CI 56% to 86%); and negative predictive value, 97% (95% CI 93% to 99%). In the 13 patients who were hypotensive, ultrasound scan correctly identified intraperitoneal fluid in all 7 patients (sensitivity 100%) with IAI, and hemoperitoneum and was negative in all 6 patients (specificity 100%) who did not have hemoperitoneum. Nine patients had IAI without intraperitoneal fluid, and ultrasound scan result was negative for fluid in all 9. CONCLUSIONS: ED abdominal ultrasound scan used solely for the detection of intraperitoneal fluid in pediatric blunt trauma patients has a modest accuracy. Ultrasonography has the best test performance in those children who are hypotensive and should be obtained early in the ED evaluation of these patients. PMID- 11431760 TI - Severe blunt hepatic trauma in children. AB - BACKGROUND: Severe blunt hepatic injury in children is associated with a high mortality rate. Although nonoperative management has become the treatment of choice for mild to moderate liver trauma, there is no consensus as to the optimal treatment for the most severe hepatic injuries in children. METHODS: A statewide trauma registry was reviewed to identify children (age 18 years or less) treated for a severe blunt liver injury for the period 1993 to 1998. Only children with an American Association for the Surgery of Trauma grade V (AIS code 541828.5) liver injury were included. Database records were reviewed for demographic information, associated injuries, survival rate, length of stay (LOS), intensive care days (ICUD), and treatment rendered after resuscitation in the emergency department. RESULTS: Thirty children with a grade V liver injury were identified. The mean age was 11.2 years (range, 1 to 18), and the overall survival rate was 56%. Data for 5 patients were excluded (4 patients died in the emergency department, and 1 patient was transferred to another institution after arrival). Survivors had a trend toward a lower injury severity score (ISS) (36.1 v 44.6; P <.1) and a significantly higher Glasgow Coma Scale (GCS), 12.5 v 6.6; P <.007). Patients with a decreased GCS had a lower overall survival rate (GCS < 8, 30% v GCS > 8, 76%). In the subset of 14 patients taken directly to the operating room, there was no difference between survivors (n = 6, 43%) and nonsurvivors (n = 8, 57%) in ISS (43 v 43; P value, not significant) or GCS (8.6 v 8.0; P value, not significant). Of the 11 patients treated nonoperatively, 10 (91%) survived with an average ISS of 33 and GCS of 13.8. Nonsurvivors more often had identified associated injuries to other abdominal and retroperitoneal organs. CONCLUSIONS: Severe hepatic injury is associated with a very high overall mortality rate in children. A low GCS is associated with a significant decrease in survival rate and may be the most important factor in outcome. Patients taken directly to the operating room have a slightly greater injury severity and a decreased survival rate compared with those treated nonoperatively. Thresholds and indications for laparotomy in these patients are not clear, and the need for operative management should be guided by the child's physiologic response to resuscitation. For those patients whose physiologic response to resuscitation permitted nonoperative management, a good outcome was achieved. PMID- 11431761 TI - Surfactant and inhaled nitric oxide in rats alleviate acute lung injury induced by intestinal ischemia and reperfusion. AB - BACKGROUND: Respiratory distress and acute lung injury (ALI) are severe complications of intestinal ischemia and reperfusion injury (I/R). The authors hypothesize that a combined surfactant and inhaled nitric oxide (iNO) may alleviate I/R-induced ALI. METHODS: Adult rats (body weight 285 to 315 g, n = 45) were allocated randomly to either a negative control group (N-Control, n = 9) with only sham laparotomy, or groups (n = 9 each) for induction of I/R by occlusion of superior mesenteric artery, followed by treatment with (1) surfactant at 100 mg/kg (Surf), (2) iNO at 20 ppm (NO), (3) both surfactant and iNO (SNO), or (4) no surfactant no iNO (a positive control, P-Control). Mechanical ventilation was provided for 120 minutes with variable peak insufflation pressure and FIO2 to achieve adequate arterial pH, PaO2, and PaCO2. Blood gas values, dynamic lung compliance (Cdyn), and airway resistance (Raw) were measured during the 2-hour treatment. Lung wet-to-dry weight ratio (W/D), histopathology, and morphometric analysis of alveolar expansion (V(V)) were determined at the end of the experiment. RESULTS: After 120 minutes of treatment, the SNO group had the highest PaO2 and Cdyn values, close to that of the N Control group. The Surf, NO, and SNO groups had higher V(V) and lower CV (V(V)) values than the P-Control, but modest decrease of W/D values. The NO group had moderately improved PaO2 but Cdyn and V(V) were as low as that of the P-Control. The lungs of the SNO group had significantly alleviated edema and neutrophil infiltration compared with the P-Control. CONCLUSIONS: The combined surfactant and iNO treatment alleviated rat ALI induced by I/R, and exerted effects better than the use of surfactant or iNO alone. PMID- 11431762 TI - Mechanisms for the development of esophageal atresia. AB - BACKGROUND: There is no universally accepted theory to explain esophageal embryology and the abnormal development that produces esophageal atresia. METHODS: The impact of Adriamycin administration on the pathogenesis of esophageal atresia was studied in the rat model of VATER association, from embryonic day (ED) 10 to ED 13. RESULTS: Tissues in the ED10 Adriamycin-exposed embryos displayed less cell proliferation as shown by the reduced population of MIB-5-labelled cells. Cell apoptosis that is characteristic of the normal ED 12 lateral epithelial folds of the foregut (the prospective site of tracheoesophageal septation) was absent in the foregut of the Adriamycin-exposed embryo. Histologic examination of the ED 11-exposed embryo showed the presence of abnormal notochord that was stretched, split, or tethered to the foregut. This contrasts with the normal embryo in which the notochord was localized in close vicinity of the ventral part of the neural tube and separated from the foregut by ample amount of mesenchyme. The abnormal localization of the notochord was accompanied by the lack of down-regulation of the sonic hedgehog (Shh) activity in the prospective site of future tracheoesophageal separation in the exposed ED 12 embryo. CONCLUSION: The authors proposed that the ectopic location of the notochord leads to the disruption in Shh signalling that may underpin the development of esophageal atresia. PMID- 11431763 TI - Changes in bladder of rabbits subjected to elevation of intraabdominal pressures. AB - BACKGROUND/PURPOSE: An experimental study has been conducted to evaluate the effects of increased intraabdominal pressure (IAP) on the morphology of the bladder of rabbits. METHODS: Experiments were performed on 20 adult male New Zealand rabbits. Six rabbits served as the control group (group I). Seven rabbits were subjected to increased IAP of 7 cm H2O for 10 days through installing air into the abdominal cavity (group II). Increased IAP was maintained for 60 days in another group of 7 rabbits (group III). Bladders were removed and fixed in 10% formalin for routine process. Paraffin sections of 5 to 7 microm were stained with H & E for light microscopic evaluation. Histopathologic parameters were scored, and the mean scores according to groups were compared by 1-way analysis of variance (ANOVA). The mean values of groups were compared separately by Tukey Kramer multiple comparison test. In these tests, P value less than.05 was considered statistically significant. RESULTS: All of the bladder strips obtained from animals subjected to 10 days of pressure increase (group II) showed mild to severe degree of vacuolation and desquamation of urothelium. Both vacuolation and desquamation of urothelium were present in all of the strips obtained from rabbits with 60 days pressure increase (group III). Additionally, there were infiltration and congestion of the urothelium together with vacuolation, suburothelial edema, and desquamation in 4 group III rabbits. Moderate or severe congestion in the lamina propria was present in bladder strips of group II rabbits. The congestion of the lamina propria was advanced, and additional moderate to severe inflammation was present in 4 rabbits of group III. Mean histopathologic scores of urothelium (P <.00001) and lamina propria (P =.002) differed significantly among groups. When the groups were compared one by one, the differences between the group I and group II and group II and III were significant (P <.05). Although serosa appeared normal in both group I and II, moderate congestion and infiltration of the serosa was present in the bladder strips of group III (P <.05). CONCLUSION: Increases in IAP for even 10 days show damaging effects on the bladder. Extended period resulted in augmentation of the damage. PMID- 11431764 TI - Infantile hypertrophic pyloric stenosis: a comparative study of pyloric traumamyoplasty and Fredet-Ramstedt pyloromyotomy. AB - PURPOSE: The aim of this study was to compare the incidence of surgical complications (duodenal perforation, postoperative vomiting, wound infection or dehiscence, incisional hernia) between 2 different surgical techniques for the resolution of hypertrophic pyloric stenosis in children. METHODS: A clinically controlled, randomized study with follow-up from 24 to 36 months was conducted. One hundred children between 15 days and 2 months old, who underwent surgical resolution of hypertrophic pyloric stenosis, were put randomly into 2 groups: I, pyloric traumamyoplasty (n = 43); II, Fredet-Ramstedt pyloromyotomy (n = 57). Both groups were controlled for the main demographic variables. Postoperative follow-up was blind for the surgical team. Statistical analysis was done with simple frequencies, percentages, Student's t test, and chi(2). RESULTS: There was not a single case of duodenal perforation, incomplete pyloromyotomy, wound infection, dehiscence, or incisional hernia in any group (P value, not significant). Postoperative emesis was present in 8 patients, uniformly distributed between groups. The operating room time for traumamyoplasty was 39.3 +/- 16.4 minutes versus 54 +/- 16.4 minutes for pyloromyotomy (P =.0003). CONCLUSIONS: This controlled study proves that traumamyoplasty is a simple procedure, quicker to perform, and as safe as pyloromyotomy for the treatment of infantile hypertrophic pyloric stenosis in children. For these reasons, the authors believe it should be considered as an alternative. PMID- 11431765 TI - Corrosive injury-induced gastric outlet obstruction: a changing spectrum of agents and treatment. AB - METHODS: During the last 5 years, 61 children were admitted to the authors' hospital because of corrosive substance ingestion, and among them 6 patients were seen with gastric outlet obstruction. Two of them had ingested acid substances, and the other 4 had ingested alkali corrosives. The mean age was 2.9 years (range, 1.5 to 3). Their common complaint was postprandial vomiting, which had begun 3 weeks after the event (range, 1 week to 10 weeks). Endoscopic evaluation and barium contrast radiographies were performed at admission. Four patients had a pyloric stricture, 1 had an antral stricture, and another had an antropyloric stricture. Balloon dilatation of the pylorus (in 1 patient), pyloroplasty (in 3 patients), and Billroth I procedures (in 2 patients) were performed. The mean follow-up period was 22 months (range, 6 weeks to 48 months). One patient, who had undergone a Billroth I procedure, underwent reoperation because of intestinal obstruction 3 months later. On follow-up they are all free of symptoms. CONCLUSIONS: The treatment of gastric outlet obstruction caused by corrosive ingestion should be treated surgically. Although endoscopic and radiologic evaluation helps to determine the time and necessity, once the diagnosis is confirmed, early definitive surgical intervention should be performed, and the type of the surgery depends mostly on the findings of the surgeon at laparotomy. Endoscopic balloon dilatation of the pylorus maybe attempted in suitable cases. Special care should be given to prevent children from accidental corrosive ingestion. PMID- 11431766 TI - A prospective study of a one-week nonbismuth quadruple therapy for childhood Helicobacter pylori infection. AB - PURPOSE: In the eradication of Helicobacter pylori infection, a 1-week therapy improves patient compliance, but drug resistance may limit its efficacy. The effectiveness of the 1-week nonbismuth quadruple therapy was studied prospectively in children with proven H pylori infection in a population with a high rate of metronidazole resistance. METHODS: All pediatric patients who presented to our institutions with acute and chronic upper gastrointestinal conditions requiring endoscopy from June 1997 to February 2000 were investigated prospectively for H pylori infection. Gastric biopsy specimens were analyzed with rapid urease test and histopathology, H pylori-positive children were treated with omeprazole, clarithromycin, amoxicillin, and metronidazole for 7 days. The result of treatment was assessed 1 month after treatment with endoscopy and biopsy. The same treatment was repeated for 2 weeks if H pylori was still present. In patients who needed a third endoscopy, their biopsy specimens were cultured to determine antibiotic sensitivity. Results were correlated with patients' symptoms and endoscopic findings. RESULTS: Thirty-three children with acute (severe epigastric pain, n = 14; gastrointestinal bleeding, n = 9) and chronic (recurrent abdominal pain, n = 7; anemia, n = 3) conditions were treated for H pylori. Thirty-one (94%) were confirmed to have H pylori eradicated by a 1 week therapy, whereas 1 patient had eradication after a further 2-we'ek therapy (3.3%). The only unresponsive patient had H pylori resistant to both clarithromycin and metronidazole. All ulcers and erosions healed after the eradication of H pylori. Three patients had persistent recurrent abdominal pain despite H pylori eradication. CONCLUSIONS: The 1-week therapy with omeprazole, clarithromycin, amoxicillin, and metronidazole is an effective treatment of H pylori in children in a population with a high incidence of metronidazole resistant strain of H pylori. Peptic ulcers and erosions healed with the eradication of the bacteria. PMID- 11431767 TI - Magnetic resonance angiography of portal collateral pathways after hepatic portoenterostomy in biliary atresia: comparisons with endoscopic findings. AB - BACKGROUND/PURPOSE: This study was undertaken to assess the usefulness of magnetic resonance angiography (MRA) in detecting varices and to investigate the possibility of avoiding routine endoscopy after surgery for biliary atresia (BA). METHODS: The subjects are 21 patients who have undergone periodical MRA since 1996. The esophageal and gastric vessels were investigated as the sites of potential development of collateral vessels. The collateral vessels on MRA were compared with endoscopic findings. RESULTS: (1) Detection of collateral vessels: of 21 patients examined, 9 patients had 12 varices. MRA depicted collateral vessels in all of the 12 varices (sensitivity, 100%). However, all MRA findings, but one was compatible with endoscopic findings (specificity, 92.9%). Outside of a series of these 21 patients, there were 3 additional patients who had undergone endoscopic treatments previously with success, and in whom esophageal vessels were not depicted on MRA, but endoscopy showed remnant varices. (2) Time dependent relationship between the appearance of collateral vessels and varices: of 4 varices that appeared after the start of MRA, 3 esophageal varices were found endoscopically simultaneously with or after delineation of collateral vessels on MRA. In the remaining patient, varices were found enodscopically 6 months before the MRA delineation, because a simultaneous MRA was not performed in this case. All these varices were in the early stage. CONCLUSIONS: MRA was highly sensitive and specific in detecting esophagogastric collateral vessels of the portal venous system. An endoscopic examination is unnecessary until collateral vessels are seen on MRA after surgery for BA. PMID- 11431768 TI - Immunohistochemistry of the liver and biliary tree in extrahepatic biliary atresia. AB - BACKGROUND: Progressive destruction of intrahepatic bile ducts may determine outcome in extrahepatic biliary atresia (EHBA) despite successful portoenterostomy. The aim of this study was to characterize the inflammatory infiltrate of a large series of cases of biliary atresia and relate these findings to clinical outcome. METHODS: Immunohistochemical analysis was performed on frozen tissue sections of extrahepatic biliary tree and liver biopsies obtained (August 1996 to March 1998) from 28 infants with EHBA and 8 liver biopsy specimens from age-matched controls with other cholestatic liver disorders. A semiquantitative scoring system was designed to evaluate the staining with a panel of antibodies to the CD4, CD8, CD25, CD56, CD68, CD71 antigens and to HLA DR, ICAM-1, VCAM-1, E-selectin and LFA-1. The infants then underwent followup prospectively and divided into 2 prognostic groups at 12 months postoperatively: those who had cleared their jaundice (graded as a good outcome [n = 19]), and those who required liver transplantation or who had failed to clear their jaundice (defined as > 50 micromol/L; graded as poor outcome [n = 9]). RESULTS: CD4(+) lymphocytes and CD56(+) (NK cells) predominated in the liver of infants with EHBA as compared with controls. The infiltrating cells exhibited marked proliferation (CD71 expression) and activation (particularly LFA-1 but also CD25 expression). A smaller subpopulation of the cells also expressed VCAM and E selectin. HLA-DR was strongly expressed on Kupffer cells and to a lesser extent on proliferating bile ducts and sinusoidal endothelium. Expression of the majority of markers was lower in the remnant bile duct tissue than in the liver of EHBA (P <.05) with only HLA-DR and LFA-1 (on infiltrating cells) and ICAM (on endothelium) expressed strongly in the remnant bile duct tissue. Although quantitatively less pronounced, all of these immunohistochemical features also were noted in non-EHBA cholestatic liver tissue. A good outcome at 12 months was associated with lower CD68 (macrophage) expression in both the liver (P <.05) and biliary tree (P <.05) and with reduced expression of ICAM-1 (P =.05) on infiltrating cells in the biliary remnant. CONCLUSIONS: Immunohistochemical patterns of immune-mediated liver injury and inflammation were prevalent features at the time of portoenterostomy. They were neither exclusive to nor characteristic of EHBA. A reduction in the expression of the macrophage marker (CD68) within the liver and biliary remnants and reduction of ICAM-1 expression on infiltrating cells in the biliary remnants appear to be associated with a better postoperative prognosis. PMID- 11431769 TI - Congenital absence of the portal vein and role of liver transplantation in children. AB - BACKGROUND/PURPOSE: Congenital absence of the portal vein (CAPV) is a subtype of congenital portosystemic shunt, which can cause a broad spectrum of clinical manifestations. The authors report on 4 patients with CAPV including a boy with CAPV-associated encephalopathy, which was resolved effectively by liver transplantation (LT). METHODS: The case records of 4 pediatric patients with CAPV who were referred to the author's institution between 1984 and 1999 were reviewed. RESULTS: The patients (3 boys and 1 girl) ranged in age at diagnosis from 0.8 to 14 years. Two patients had growth retardation or disturbed consciousness, and the other 2 had no specific manifestations. Not only high serum levels of bile acids, ammonia, and transaminases but also low plasma levels of branched-chain amino acids were common laboratory findings. The absent portal vein was replaced by a large portosystemic shunt, which connected the splanchnic vein to the inferior vena cava or the left renal vein. Two patients survived without any symptoms, but 1 with growth retardation died of hepatic failure. The other with encephalopathy did not respond to medical therapy and underwent LT, which resolved symptoms and metabolic disorders effectively. CONCLUSIONS: Patients with CAPV do not always have a good prognosis. They should be followed up with careful observation of their symptoms, hepatic function, and metabolic abnormalities. LT might be indicated for patients with symptomatic CAPV unresponsive to medical therapy. PMID- 11431770 TI - Sodium cromoglycate in the management of chronic or recurrent enterocolitis in patients with Hirschsprung's disease. AB - BACKGROUND/PURPOSE: Chronic or recurring enterocolitis is a rare but perplexing complication of Hirschsprung's disease affecting especially patients with altered immune defense such as those with Down's syndrome. Sodium cromoglycate (SCG) is a nonabsorbable mast cell stabilizing agent that has been documented to be effective in the treatment of inflammatory bowel disease. The authors studied the effect of SCG in Hirschsprung patients with refractory chronic or recurrent enterocolitis. METHODS: Eight patients (4 with Down's syndrome, 2 with other chromosomal aberrations, 2 otherwise healthy; age range from 4 to 22 years) with chronic (5 patients) or recurrent (>6 episodes/year, 3 patients) enterocolitis received 100 to 200 mg of SCG 4 times a day depending on the age of the patient. The chronic diarrhea or recurrent bouts of enterocolitis in the patients were refractory to dietary management and enteral antibiotics. Before the treatment all patients had ileocolonoscopy, the results of which showed macroscopic and histological chronic inflammation in all cases. No neuronal abnormalities were detected in biopsy results. None of the patients had colonic dilatation or increased anorectal resting pressures suggesting outlet obstruction. RESULTS: The follow-up of the patients ranges from 8 months to 26 months. Three of the 5 patients with chronic enterocolitis responded favorably. In these 3 patients the median number of daily bowel movements decreased from 6 to 3, and none experienced bouts of abdominal distension. Diarrhea-related soiling decreased also significantly. Two of the 3 patients with recurrent enterocolitis have remained asymptomatic, and none has required antibiotics after the onset of SCG treatment; one patient had an episode of enterocolitis after 12 months treatment. Two patients with chronic enterocolitis did not respond to SCG. No side effects of SCG were encountered. CONCLUSIONS: This preliminary and nonrandomized study suggests that SCG is an effective treatment modality for chronic or recurrent enterocolitis in patients with Hirschsprung's disease. Because SCG is not absorbed From the intestinal tract there are no systemic side effects. PMID- 11431771 TI - Dynamic graciloplasty in children with fecal incontinence: a preliminary report. AB - BACKGROUND: Various therapeutic methods have been developed for anal incontinence. A reconstruction of the sphincter system with functional adaptation can be achieved by stimulation of the transposed gracilis muscle with an implanted stimulator. METHODS: The authors performed a dynamic graciloplasty in 4 boys, aged between 6 and 10 years. Causes for the incontinence were polytrauma with pelvic rupture, VACTERL syndrome with rectal urethral fistula, anal atresia with primary reconstruction and 6 consecutive operations, and coccygeal teratoma in a premature infant with surgical treatment of the recurrent tumour. All children had grade III incontinence. Postoperatively, the duration of stimulation was increased successively by telemetric programming of the pulse generator up to a continuous mode. RESULTS: One child is now almost continent, 2 show grade I incontinence, 1 is incontinent with frequent soiling (the programming has not been completed yet). CONCLUSION: An evaluation of this method for children will not be possible until more operations have been performed, under the conditions of a prospective study, which appears indicated in view of the preliminary but encouraging results presented. PMID- 11431772 TI - Expression of matrix metalloproteinases and the metastasis-associated gene S100A4 in human neuroblastoma and primitive neuroectodermal tumor cells. AB - BACKGROUND: Matrix metalloproteinases (MMPs) and their endogenous inhibitors (tissue inhibitors of MMPs; TIMPs) have been shown to correlate with in vitro invasiveness and clinical outcome in several adult malignancies. The importance of MMP and TIMP expression in neuroblastoma (NB) and primitive neuroectodermal tumors (PNET) is incompletely understood. The aim of the current study was to relate in vitro invasion of NB and PNET cell lines with MMP and TIMP expression and evaluate the effect of a synthetic MMP inhibitor. Furthermore, S100A4 levels were determined because recent reports have suggested a possible association between MMPs, TIMPs, and the metastasis-associated gene S100A4. METHODS: Expression of MMPs, TIMPs, and S100A4 was evaluated at both mRNA and protein levels in 2 human NB and 2 PNET cell lines. In vitro invasion and effects of the synthetic MMP inhibitor Marimastat were assessed in the Transwell chamber assay. RESULTS: The most invasive cells expressed the highest levels of MMPs and S100A4. Marimastat reduced invasion by 30%. CONCLUSIONS: In vitro invasion correlated with MMP and S100A4 expression. The fact that Marimastat reduced in vitro invasion is encouraging for further studies on a possible therapeutic application for proteinase inhibitors. PMID- 11431773 TI - The normal position of the umbilicus in the newborn: an aid to improving the cosmetic result in exomphalos major. AB - BACKGROUND/PURPOSE: The aim of this study was to determine the normal position of the umbilicus and hence improve the cosmetic result of exomphalos major repair. METHODS: The position of the umbilicus was determined in 50 neonates with respect to the xiphisternum and pubis. RESULTS: The normal umbilical position is 60% of the way from the inferior border of the xiphisternum to the superior border of the pubis in the midline. CONCLUSION: In the repair of exomphalos major, the most aesthetically pleasing result is obtained if the umbilicus is placed 60% of the way from the xiphisternum to the pubis. PMID- 11431774 TI - Gomco circumcision: When is it safe? AB - BACKGROUND/PURPOSE: The Gomco clamp is used most commonly for neonatal circumcisions in the United States with reported rates of complication as low as 0.2%. Often, however, circumcision is delayed beyond the neonatal period because of illness, parental concerns, or physician bias with patients presenting for elective circumcision in the first few years of life. In neonates and infants, the "bloodless" Gomco circumcision is cost effective, can be performed with minimal anesthesia and gives good cosmetic results. However, the optimal age at which to perform this procedure remains controversial. We reviewed the experience of one pediatric urologist with Gomco circumcision in neonates and infants to determine the optimal age at which this procedure should be performed. METHODS: One hundred thirty patients underwent Gomco circumcision by 1 pediatric urologist between 1996 and 1998. Ninety-eight patients underwent Gomco circumcision as neonates or during early infancy at a mean age of 17 days (range, 4 to 30 days). Thirty-two patients underwent circumcision after early infancy at a mean age of 6.5 months (range, 90 days to 8.5 months). All patients younger than 1 month of age underwent circumcision while fully awake using a dorsal penile nerve block. Patients greater than 3 months of age were circumcised under general anesthesia. Gomco circumcision was performed using 1.1- to 2.1-cm bells. Patients were assessed with regard to outcome. RESULTS: None of the 98 patients in the early infancy group had postcircumcision complications. Of the 32 patients in the older group, 12 (30%) had postoperative bleeding requiring suture repair or fulguration. The skin edges separated in 2 patients, 1 less than 1 month of age and 1 in the older group. Neither patient required further intervention, and both healed spontaneously. There was no correlation between size of clamp and development of complications. CONCLUSION: Although safe and effective for circumcision in the neonatal period and in early infancy, use of the Gomco clamp for circumcision beyond early infancy (3 months of age) has substantial morbidity, and alternative methods of circumcision should be sought. PMID- 11431775 TI - Evaluation of contralateral testicular damage after unilateral testicular torsion by serum inhibin B levels. AB - BACKGROUND/PURPOSE: It is still controversial whether unilateral testicular torsion (TT) affects contralateral testis. The authors wanted to evaluate contralateral testicular damage in a rat model by the serum inhibin B levels, which is suggested as a marker of Sertoli cell function and spermatogenesis. METHODS: Fifty peripubertal male Wistar Albino rats were divided into 5 groups each containing 10 rats. Surgery was conducted under intraperitoneal 1-shot ketamine (50 mg/kg) anesthesia. Torsion-detorsion, torsion-detorsion orchiectomies, orchiectomies, and sham operations were performed on the right testicle through a midline incision. Torsions were created by rotating the right testes 720 degrees in a clockwise direction and maintained by fixing the testes to the scrotum with a silk suture. Torsion duration was 4 hours. After each surgical intervention the incisions were closed. In group 1, 3-mL blood samples were taken to determine basal values of inhibin B in serum, and bilateral orchiectomies were performed. In group 2, 4 hours of torsion and detorsion was created and 1 month later, 3-mL blood samples were taken, and bilateral orchiectomies were performed. In group 3, 4 hours of torsion-4 hours of detorsion was created, and right orchiectomies were performed and 1 month later, 3-mL blood samples were taken and contralateral orchiectomies were added. In group 4, unilateral orchiectomies were performed, and 1 month later, 3-mL blood samples were taken, and contralateral orchiectomies were added. After the measurement of the serum inhibin B levels and histopathologic examinations, results are expressed as mean +/- SD. RESULTS: Serum inhibin B levels expressed as mean +/- SD were 108.233 +/- 21.296 pg/mL for group 1, 54.065 +/- 16.910 pg/mL for group 2, 74.195 +/- 2.779 pg/mL for group 3, 108.335 +/- 26.078 pg/mL for group 4, and 107.645 +/- 4.705 pg/mL for group 5. Inhibin B levels in group 2 and group 3 were different from group 1, group 4, and group 5 (P <.05). In histologic examination, Johnsen's scores expressed as mean +/- SD of right testes were 9.74 +/- 0.08 for group 1, 3.64 +/- 3.36 for group 2, and 9.86 +/- 0.05 for group 5. Histologic findings in group 2 were different from group 1 and group 5 (P <.05). Johnsen's scores expressed as mean +/- SD of left testes were 9.78 +/- 0.09 for group 1, 9.75 +/- 0.14 for group 2, 9.76 +/- 0.15 for group 3, 9.79 +/- 0.07 for group 4, and 9.82 +/- 0.08 for group 5, and there was no difference between groups (P >.05). CONCLUSIONS: The serum inhibin B levels decrease after unilateral TT reflecting contralateral testicular damage. Orchiectomy to prevent contralateral testicular damage after TT may not be effective after critical period. Measurement of inhibin B levels to evaluate contralateral testicular damage after unilateral TT is more effective than histopathologic examination. PMID- 11431776 TI - Coccidioides immitis in the gallbladder and biliary tree. AB - Intraabdominal coccidioidomycosis is a very rare entity and usually responds to medical therapy. Operative intervention is reserved for diagnosis or drainage of localized collections. With biliary coccidioidomycosis, medical treatment appears to be ineffective, and biliary tract drainage is necessary for optimal management. A case of coccidioidomycosis in the gallbladder and biliary tree is described and the literature reviewed. PMID- 11431777 TI - Diffuse fibrous proliferation of tunica vaginalis associated with testicular infarction: a case report. AB - Fibrous pseudotumor of the tunica vaginalis testis is an uncommon lesion of unknown pathogenesis. Although this reactive process of testicular tunics is benign, this usually is diagnosed after radical orchiectomy. The authors describe a case of fibrous pseudotumor of the tunica vaginalis testis associated with testicular infarction. To our knowledge, this is the first case presented with similar association, second case consisting predominantly of myofibroblasts and the fourth reported case encountered in childhood. PMID- 11431778 TI - Peritoneal encapsulation: a rare cause of bowel obstruction in children. AB - Two rare cases of peritoneal encapsulation causing partial intestinal obstruction are described. Both were operated on, and one of them had a "cocoonlike" appearance. The characteristics and differences of peritoneal encapsulation and abdominal cocoon are discussed. PMID- 11431779 TI - Primary pulmonary rhabdomyosarcoma arising within cystic adenomatoid malformation: a case report and review of the literature. AB - The current report describes a 13-month-old boy with primary pulmonary rhabdomyosarcoma (RMS) that originated within a congenital cystic adenomatoid malformation (CCAM). To the best of our knowledge, he is the youngest patient of all primary pulmonary RMS cases reported in the English-language literature. The tumor, localized in left upper lobe, was removed completely, and histologic examination showed embryonal subtype. Postoperative systemic chemotherapy was carried out. Recent evaluation 15 months after resection has not identified any residual or recurrent disease. Primary pulmonary RMS, although very rare in the pediatric age group, should be considered in young patients with solitary pulmonary masses and associated cystic lesions. PMID- 11431780 TI - Delayed neutropenic enterocolitis in a 12-year-old girl treated with total colectomy and J-pouch reservoir. AB - Neutropenic enterocolitis (NE) is a clinicopathologic condition characterized by bowel wall inflammation, which can proceed to necrosis and perforation. It is mostly seen in neutropenic patients with leukemia who undergo induction treatment with chemotherapy. Most often the cecum is involved. The authors present a 12 year-old girl with acute lymphocytic leukemia who, under maintenance therapy, experienced NE. The disease was localized to the left side of colon, and even the rectum was involved, which is an unusual localization of the disease. An ileoanal anastomosis with a J-pouch was done in a second operation with a good outcome. PMID- 11431781 TI - Female circumcision. AB - Female circumcision, more accurately known as female genital mutilation, is still a common practice in parts of Africa. This ritual genital operation can involve partial or complete excision of the clitoris and labia minora as well as infibulation (labial fusion). The case reported here involves a 16-year-old African girl with a complication of this mutilating procedure. PMID- 11431782 TI - Vaginal reconstruction using a vertical rectus abdominis myocutaneous flap in a child: a case report. AB - Rhabdomyosarcoma is the most common soft tissue sarcoma in infants and children younger than 15 years old. Multimodal treatment using chemotherapy, radiation, and surgery have increased survival. Because of the potential for long-term survival, the importance of organ preservation and vaginal and pelvic reconstruction has been recognized. The authors report a case in which special circumstances led to vaginal reconstruction concomitantly with exenteration surgery. The authors present the first reported use of a vertical rectus abdominis myocutaneous flap in a child and describe the technique and its potential advantages. PMID- 11431783 TI - Duodenojejunal intussusception secondary to hamartomatous polyps of duodenum surrounding the ampulla of Vater. AB - Duodenojejunal intussusception is a rare pediatric emergency. A case of duodenojejunal intussusception secondary to hamartomatous polyps of the second portion of duodenum in a 10-month-old boy is reported. Surgical excision of the polyps and reduction of the intussusception were performed. Pathologic examination found hamartomatous polyps. This is the third case report of children in literature, but this is the first case of a child with intussusception surrounding the ampulla of Vater and a successful excision performed without damaging the ampulla of Vater. PMID- 11431784 TI - Perineo-pelvic diaphragmatic split: anorectal exstrophy. AB - Anorectal malformations are among the most common congenital anomalies that require neonatal surgery. Although the headlines of anorectal malformations present with a spectrum of defects of this region, generally, imperforate anus and its fistulous or nonfistulous connections are examined. Here the authors present a boy with anorectal malformation, with sacrococcygeal teratoma and meningocele. Interestingly, the patient had neither imperforate anus nor anorectal stenosis, but had widely open anorectum. This kind of anorectal malformation has never been described previously in the English-language literature. PMID- 11431785 TI - Neonatal sigmoid volvulus: a complication of anal stenosis. AB - Sigmoid volvulus is an exceptionally rare cause of intestinal obstruction in neonates. Only 7 cases have been reported in the English and French literature. The authors report a recent case of sigmoid volvulus in a neonate secondary to anal stenosis and review the diagnosis and management of this serious condition. The authors believe that carefully performed radiologic reduction is the preferable alternative to surgical intervention. PMID- 11431786 TI - A critical observation about the pathogenesis of abdominoscrotal hydrocele. AB - Abdominoscrotal hydrocele is a very rare anomaly. Although various theories about this condition have been proposed, controversy still continues on the etiology of this lesion. The authors present two consecutive cases of abdominoscrotal hydrocele in infancy. On their first examination, these patients only had inguinoscrotal hydroceles. After a 2-month period of observation, these inguinoscrotal hydroceles developed into abdominoscrotal hydroceles. These findings also were confirmed by ultrasonography. The authors question the currently proposed theories of abdominoscrotal hydrocele development. PMID- 11431787 TI - Ectopic liver tissue within the umbilical cord. AB - A solid mass in the umbilical cord is a very rare finding. A neonate with isolated ectopic liver tissue found in the umbilical cord is reported. This is the first known such reported case. PMID- 11431788 TI - Complex branchial fistula: a variant arch anomaly. AB - A 5-year-old boy presented with an infected left-sided branchial fistula. Despite antibiotic treatment and repeated excision of the fistula, purulent discharge from the wound persisted. Three-dimensional computed tomography (3D CT) reconstruction greatly facilitated the diagnosis and management of this case by showing the course of the fistulous tract. The complexity of the tract suggests that this represents a variant arch anomaly because it contains features of first, second, third, and fourth arch remnants. PMID- 11431789 TI - Splenosis mimicking tumor recurrence in renal cell carcinoma: detection on selective spleen scintigraphy. AB - A 2-year-old boy had been operated on for a giant renal cell carcinoma including splenectomy because of disrupture of the splenic capsule. During a follow-up examination, 3 nodules were detected by ultrasound in the splenorenal area. This gave reason to suspect tumor recurrence. Considering the possibility of splenosis, a selective spleen scan using denatured red blood cells was performed as a final diagnostic step. This method confirmed the nodules as representing splenic tissue. Splenosis should be included in the differential diagnosis of solid masses in the postsplenectomy patient. PMID- 11431790 TI - Management of accessory hepatic ducts in choledochal cysts. AB - This report describes the surgical management of 2 children with fusiform choledochal cysts who had accessory hepatic ducts (AHD) identified during excisional surgery for fusiform choledochal cysts (CC). Two children presenting with a triad of recurrent jaundice, fever, and abdominal pain were investigated and found to have type 1 choledochal cyst. Preoperative imaging and intraoperative cholangiography missed the AHD in both cases. In one of the patients, the main and the accessory ducts were separated by the right hepatic artery. In both the patients the accessory ducts were reconstructed successfully into a Roux loop along with the main common hepatic duct. Follow-up studies showed no evidence of biliary tract obstruction or atrophic changes in the liver. There was satisfactory uptake and drainage on hepatic scintigraphy. During excision of CC, AHD may be encountered. These may be missed on preoperative imaging. AHD may have a close relationship with neighboring vascular structures in the porta. Accessory hepatic ducts should be anticipated, identified, and reimplanted into the Roux loop during excisional surgery. PMID- 11431791 TI - Acute symptomatic hyponatremia and cerebral salt wasting after head injury: an important clinical entity. AB - Hyponatremia is a well known complication of traumatic and nontraumatic cerebral injury, often related to the syndrome of inappropriate antidiuretic hormone secretion (SIADH). Nonetheless, it also can be associated with a different entity, the syndrome of cerebral salt wasting (CSW). The authors report the case of a 4.5-year-old boy presenting with major head injury who at day 6 after admission had generalized tonic-clonic seizures caused by severe acute hyponatremia (serum sodium level, 119 mmol/L) and signs of dehydration. Despite initial isotonic rehydration, hyponatremia persisted because of excessive renal salt losses and concomitant enormous water losses, necessitating increasing amounts of sodium, up to 160 mmol/kg/d, and large amounts of intravenous fluids, up to 27 L/d. Highly increased levels of atrial natriuretic peptide (ANP) confirmed the diagnosis of CSW. The occurrence of a CSW has to be recognized early in the clinical course for adequate treatment and remains one of the important differential diagnosis of SIADH in hyponatremic states in patients with cerebral disorders, especially after head injury. PMID- 11431792 TI - Intestinal atresia associated with twin-twin transfusion syndrome. PMID- 11431794 TI - Female pseudohermaphroditism associated with cloacal anomalies: faulty differentiation in the caudal developmental field. AB - A 1(1/2)-year-old girl with a urogenital sinus defect associated with an enlarged phalluslike clitoris is described. The child had an anteriorly located anus, a single kidney with grade III vesicoureteric reflux, a 3.5-cm-long common urethrovaginal channel, a large vesical calculus, and a hamartoma in the pubic region. After a preliminary colostomy, definitive surgery consisted of urethral reconstruction, vaginal reconstruction using a colonic graft, and posterior relocation of the anus. The discussion centers on a review of the previously reported cases of female pseudohermaphroditism associated with cloacal or urogenital sinus defects (FPHCD). The likely embryogenesis of this condition is discussed, and the similarity of the features with those seen in some patients with complete penoscrotal transposition (CPST) is highlighted. PMID- 11431795 TI - Dysgerminoma associated with hypercalcemia: A case report. AB - Hypercalcemia is an extremely rare paraneoplastic manifestation of malignancy in children. The authors report a case of dysgerminoma in a 14-year-old girl associated with severe hypercalcemia that only resolved after excision of the tumor. There are only 6 previously reported cases of dysgerminoma associated with hypercalcemia. Serum calcium levels should be checked in all children with solid ovarian tumors. Hypercalcemia in such circumstances may be resistant to medical management, and surgical resection of the tumor results in the normalization of calcium levels. PMID- 11431796 TI - Epigastric heteropagus twinning. AB - Conjoined twins is a very rare condition that represents, sometimes, a real challenge for pediatric surgeons. There is an even rarer situation, called heteropagus twinning, when one of the twins is not completely formed (parasite) and dependent of the well-formed one (autosite). The authors present a case of heteropagus twins in which the parasite was attached to the epigastrium of the autosite. In this situation, the separation procedure may be quite simple if a well-done anatomic study using x-ray, ultrasonography, computed tomography (CT) scan, magnetic resonance, and echocardiogram are performed. The precise knowledge of the anatomic variability and physiologic changes of both twins will help the surgeon make decisions like which surgical approach to use and the dissection tactic to be used. The pediatric surgeon also must be aware of the necessity to improvise during the operation. PMID- 11431797 TI - Complete diphallia associated with features of covered exstrophy. AB - This report describes a 1-day-old boy with complete true diphallia associated with features of covered or pseudo-exstrophy and an anorectal malformation. The urinary bladder and rectum were single. A divided sigmoid colostomy was constructed at birth for the anorectal anomaly. Complete diphallia and its likely embryogenesis are discussed. PMID- 11431798 TI - Benign symmetric lipomatosis with myelomeningocele in an adolescent: An uncommon association-case report. AB - The authors report an unusual localization of symmetric adipose tumors associated with spinal dysraphism. Initially, the patient underwent a closure of the lumbosacral myelomeningocele. At that time, the tumors were not evident and remained undiscovered up until puberty. However, during puberty, the symmetric lipomatous masses grew at the perineal region. Except the patient's age, all findings and the clinical picture supported the diagnosis of a benign symmetric lipomatosis (BSL). To the authors' knowledge, the following case has not been described previously. PMID- 11431799 TI - Retrovesical echinococcal cyst causing renal failure and nocturnal enuresis in a child. AB - Although retrovesical hydatid cysts are observed in adulthood albeit very rarely, to our knowledge there is no case reported in childhood. In the current case, a huge retrovesical hydatid cyst caused obstructive uropathy, polydipsia, and secondary nocturnal enuresis in a child. Because the retrovesical hydatid cyst was firmly adherent to the neighboring structures, partial cystectomy could be performed. Perioperative and postoperative complications did not occur, and the symptoms relieved in the early postoperative period. PMID- 11431800 TI - Oromandibular limb hypogenesis and gastroschisis. AB - The oromandibular limb hypogenesis syndromes (OLHS), which are congenital malformations involving the tongue, mandible, and the limbs are extremely rare with a few sporadic cases reported in the literature. In this report, the authors report a case of OLHS with gastroschisis, that has not been described previously according to the medline search. PMID- 11431801 TI - Progress in mitral and aortic regurgitation. AB - Over the past 15 years there has been rapid and dramatic change in the therapy for valvular heart disease. When mitral and aortic regurgitation are severe, they inevitably cause left ventricular damage, eventually resulting in death. However, when surgical correction of these lesions is timed appropriately, longevity can approach that of a normal population after surgery. As surgical techniques have improved, surgery is now indicated earlier in the course of these diseases. It is clear that some patients with mitral and aortic regurgitation require surgery even though they are entirely asymptomatic. However, it must be emphasized that mitral and aortic regurgitation are quite different from one another. These different lesions result in different loading conditions, different pathophysiologies, and have different means for surgical correction. All of these issues impact on the proper timing of surgery and are discussed. PMID- 11431802 TI - Timing of surgery in aortic stenosis. AB - In adults with valvular stenosis, the importance of prompt aortic valve replacement once symptoms occur is well known. The operative mortality for aortic valve replacement has improved dramatically over the past 4 decades and remains the only effective therapy for severe symptomatic aortic stenosis. Aortic valve replacement in patients with left ventricular dysfunction has a high operative mortality, although those patients who do not undergo surgery at all have an even worse outcome. While issues to consider include the presence or absence of coronary artery disease and expected hemodynamics of the prosthetic valve compared with the native valve, when in doubt, one should err on the side of surgical intervention. Elderly age is not a contraindication to aortic valve replacement for severe symptomatic aortic stenosis, although there is a higher prevalence of comorbid disease and higher operative mortality. Life expectancy is significantly prolonged and quality of life is significantly improved in the elderly who survive surgery. Indications for surgery in asymptomatic patients are controversial. We do not recommend valve replacement in asymptomatic patients at this time due to the known risks of surgery and a prosthetic valve and the lack of evidence for benefit of early surgery. Patients undergoing coronary bypass surgery should be considered for concomitant aortic valve surgery for moderate aortic stenosis that is expected to progress to severe stenosis in less than 5 years. PMID- 11431803 TI - The internodal pathways of the human heart. PMID- 11431804 TI - Contemporary endodontology. AB - Clinical endodontics has made significant progress during this century. This lecture will critically review the major breakthroughs that have taken place during this period. The final judgement of the value of this progress, however, is the quality of care we can deliver to our patients. Although the advances have been impressive, there are also major disappointments and problems that need to be recognised. Looking at clinical endodontics today, there is clearly a dissonance between sound scientific knowledge and empirical clinical practice. It is important that these problems are recognised and that endodontic research and teaching are refocused. Only by decisive action can progress be assured for clinical endodontics and the prospect will be bright. PMID- 11431805 TI - The Gow-Gates mandibular block: regional anatomy and analgesia. PMID- 11431806 TI - The Gow-Gates mandibular block: applied anatomy and geometry. PMID- 11431807 TI - Discuss the use of local anaesthetic for endodontic treatment. PMID- 11431808 TI - Apexification with calcium hydroxide--when should the dressing be changed? The case for regular dressing changes. PMID- 11431809 TI - Cracked tooth syndrome. PMID- 11431810 TI - Early protection of cracked teeth. PMID- 11431811 TI - The 8th Biennial Congress of the European Society of Endodontology Goteborg, Sweden, June 12-14, 1997. PMID- 11431812 TI - Dentist discovers ether anaesthesia: the 150th anniversary. PMID- 11431813 TI - Dental accident insurance. PMID- 11431814 TI - The evolution of modern endodontics. PMID- 11431815 TI - Pulpal injury: pathology, diagnosis and periodontal reactions. AB - Diagnosis of pulpal disease can be difficult due to the lack of diagnostic signs and symptoms available to the practitioner. An understanding of the possible underlying pathological processes, combined with an exact assessment of the pain history, and appropriate clinical tests, should aid the practitioner in determining the nature of pulpal inflammation, and differentiating it from dentine sensitivity and cracked teeth. The responses of the pulp to traumatic injury to the periodontal membrane (PDM) require special consideration, particularly with respect to the assessment of pulp vitality, and the determination of cases requiring pulp extirpation in order to avoid inflammatory root resorption. Although the pulp is relatively isolated from the rest of the dentoalveolar complex by a dentine/cementum barrier, it is important to remember that it can communicate with the PDM through apical and lateral foramina, and areas of damaged cementum. Hence, it is a priority to both preserve the integrity of the cemental layer in cases of traumatic injury and periodontal disease, and to prevent the inflammation and resorption associated with periapical lesions by accurate diagnosis of irreversible pulpitis and pulp necrosis, followed by appropriate endodontic debridement procedures. PMID- 11431816 TI - Trauma quiz. Terribly troublesome, trauma teeth. AB - Unfortunately, we come across many traumatised teeth during our practising career. Some of these traumatic injuries are rather simple to treat whereas others provide us with a real challenge. It is absolutely essential that the diagnosis of the injury be known before any treatment is attempted. When it comes to trauma, however, defining the exact form of treatment can often be very difficult. In this paper I will discuss some of the cases that I have managed and leave it up to YOU to decide whether my treatment has been correct, incorrect or whether there is some other form of treatment that we have at our disposal that could have been attempted. PMID- 11431817 TI - The endodontic management of a late complication of intrusive luxation injury. PMID- 11431818 TI - Intentional replantation. PMID- 11431819 TI - "In the eye of the beholder"--how radiographic image interpretation depends on who is looking and when. PMID- 11431820 TI - Treatment of dens invaginatus in supernumerary teeth. AB - The endodontic treatment and aesthetic restoration of a case of Oehlers Type I dens invaginatus in supernumerary teeth is presented. PMID- 11431821 TI - Above all do no harm. PMID- 11431822 TI - Historical Glimpses. The X Factor. PMID- 11431823 TI - Pulp protection under restorations--do you need a liner? PMID- 11431824 TI - Sealing ability of furcation perforation repair. PMID- 11431825 TI - Surgical management of a tooth with stripping perforation. PMID- 11431826 TI - Hardness and strength of endodontic instruments made from NiTi alloy. AB - Scant information could be found in the dental literature on the hardness or strength of the materials in nickel-titanium (NiTi) root canal instruments. Hardness testing is a convenient method for examining these products and strength properties can be deduced from strength-hardness data. The Vickers Hardness was determined for NiTi root canal files and reamers from five manufacturers for sizes 15, 25 and 40. Significant differences in hardness were found between manufacturers, with values ranging from 296 to 349. Some differences were found between the different sizes from each manufacturer, but these were generally not statistically significant. These values obtained for NiTi instruments are lower than the softest stainless steel files previously examined (296-347 cf. 403-651), and below the hardness required for many cutting instruments (550-650). The values obtained suggest that NiTi instruments may not be as sharp as, and become blunt more easily than, stainless steel instruments. The strengths are expected to be in the lower range of those for stainless steel instruments. PMID- 11431827 TI - Cleaning the curved root canal--mission impossible? PMID- 11431828 TI - When all else fails.... AB - Endodontic surgery may be indicated if orthograde endodontic treatment cannot be carried out or is unsuccessful. Advances in instruments, materials and techniques have made surgical endodontics a more predictable procedure. A complex case that required surgical intervention is presented to illustrate this point. PMID- 11431829 TI - Vertical root fracture of a non-endontically treated tooth. PMID- 11431830 TI - Conservative treatment of an unusual sinus presentation. PMID- 11431831 TI - Patient concern: does it cease after the office closes? PMID- 11431832 TI - Making the choice. The pros and cons of outsourcing. PMID- 11431833 TI - Furniture facts. Institutional is out, aesthetics are in. PMID- 11431834 TI - What's the frequency, ASHE? Get your medical telemetry equipment registered. PMID- 11431835 TI - Where are the robots? Good and bad news for automated cleaning. PMID- 11431836 TI - Scanning on thick ice. PMID- 11431837 TI - The staff shortage: dispatches from the listserv. PMID- 11431838 TI - Ultrasound aboard the international space station. PMID- 11431839 TI - Knowledge is power. PMID- 11431840 TI - Inside interim management. PMID- 11431841 TI - Reinvent the R.T. workplace. PMID- 11431842 TI - R.T. shortage: one college's search for answers. PMID- 11431843 TI - X-ray technologists: your best investment. PMID- 11431844 TI - Recruitment & retention in seven easy steps. PMID- 11431845 TI - What does eCommerce offer the radiology department? PMID- 11431846 TI - From R.T. to entry-level manager: how to climb the clinical ladder. AB - Modern healthcare institutions are faced with an environment that is rapidly changing. Many imposed changes have repercussions that affect lower-level radiology managers. According to one expert, radiology administrators face two dilemmas in preparing for the future: upgrading skills of those currently employed and ensuring that future employees are prepared to succeed. This paper examines the knowledge, skills and abilities needed for entry-level managers in radiology to succeed in healthcare. In order to gain current information on processes used in promoting staff technologists to management positions, the authors developed and mailed a questionnaire to 35 radiology administrators across the country. In addition to promoting employees, organizations should take a role in the maintenance and growth of an employee's career by creating realistic career paths for advancement, including programs in mentoring, assessment centers or formal training in the art of interviewing candidates. In order for entry-level radiology managers to succeed in a complex healthcare environment, mere technical expertise is not sufficient. The responsibility for identifying and providing appropriate education and training lies with employers, educational programs, professional groups and the employee. PMID- 11431847 TI - Mammography technology overview. AB - As you can see, there are many options in mammography units. This article is not meant to recommend one unit or manufacturer over another, but rather to inform consumers that all units have assets to help technologists acquire the perfect mammogram. This article is meant to make you think about the pros and cons as you consider purchasing new mammography equipment. Though it may seem wise and certainly easier to equip your multiple-unit site(s) with one manufacturer's equipment or even one model, it might truly be wise to have a little variety; this addresses shortcomings of a particular unit. If you provide technologists with a mammography system with which they are comfortable and feel they can address any issues-accommodating disabled patients, patients with breast implants or patients with dense breast tissue-you will see the perfect mammograms you desire. PMID- 11431848 TI - AHRA Staff Utilization Survey highlights. PMID- 11431849 TI - Keeping the faith. PMID- 11431850 TI - [Molecular genetics of Wilms' tumor]. AB - Molecular genetics of the Wilms' tumor plays an important role in the elucidation of the genetic etiology of the tumor disease generally. Contrary to the genesis of retinoblastoma, where a single gene is inactivated by two hits, the biological signalling pathways determining the origin of the Wilms' tumor are more complex and several genes in several loci may participate. Formation of the Wilms' tumor is accompanied with the most frequent genetic alteration, which is the loss of heterozygosity on the short arm of chromosome 11. It indicates inactivation of one or several tumor suppressor genes located at 11p region. The most studied gene of the Wilms' tumor is WT1 gene, which has been cloned and sequenced. Biological function of WT1 protein is complex one and it requires probably an interaction with other proteins, DNA and also RNA. The development of the tumor determines not only the genetic changes, but also epigenetic changes, e.g., hypermethylation of promoter and genome imprinting. PMID- 11431851 TI - [Radical radiotherapy of prostate carcinoma]. AB - Radiotherapy is an effective treatment modality of prostate carcinoma. Considering the limited radiosensitivity of this tumour it is necessary to use relatively high doses of radiation. Modern technology offers the possibility to perform high-dose radiotherapy with acceptable toxicity. The authors present various options of both external and internal irradiation. They point out that brachytherapy, previously considered less appropriate from the radiobiology aspect, is according to the current views and results a very suitable treatment modality of prostate cancer. PMID- 11431852 TI - [Plasma level of Lp(a) in patients with myocardial infarct is not controlled by the apolipoprotein E gene polymorphism]. AB - BACKGROUND: We evaluated a two risk factors of atherosclerosis development, apolipoprotein E genotype and plasma level of Lp(a), and their association in Czech population. PATIENTS AND CONTROLS: Apo E and Lp(a) have been determined in the group of 109 men with premature myocardial infarction and compared with the population samples (301 men for apo E polymorphism and 112 men for Lp(a) evaluation). METHODS AND RESULTS: Apo E gene polymorphism was analysed by PCR with subsequent restriction analysis of the PCR product. Lp(a) concentration was measured enzymatically using the IMMUNO set. The frequency of alleles of apo E genotype in myocardial infarction patients did not significantly differ from that found in the control group. The distribution of the Lp(a) concentrations in MI patients was shifted to higher levels than in population sample (p < 0.0001). CONCLUSIONS: We have not found correlation between this two risk factors of myocardial infarction. High concentration of Lp(a) is on apo E polymorphism independent risk factor of development of myocardial infarction. PMID- 11431853 TI - [Ethical problems in randomized clinical trials from the aspect of statistics. Part I. Is patient randomization acceptable from the ethical viewpoint?]. AB - Randomized clinical trial is currently thought to be the most reliable method for comparing therapeutic effects. The article tries to answer the question whether chance allocation of patients into various therapeutic arms is acceptable from the ethical point of view. It deals with the conflict between two concepts: of individual ethics mostly defending interests of individual participants of each trial, and collective ethics based on well-designed experiments that are the sole source of reliable estimation of therapeutic effect and their conclusions can change clinical practice and help patients in the future. The article suggests this possible solution: providing there is no prevailing consensus as for superiority of one of the compared therapies, the offer to participate in randomized clinical trial is not only acceptable but necessary in view of individual patients' right and respect of their autonomy. If an informed consent is obtained the principles of medical ethics have not been violated. Finally, a few alternative trial designs are presented. PMID- 11431854 TI - A 7-year-old girl with failure to thrive and progressive bowing legs. PMID- 11431855 TI - Intussusception in childhood. PMID- 11431856 TI - The developments of mass urinary screening in Taiwan. AB - A program of mass urinary screening of elementary and junior high school students has been in operation since August 1990 in Taiwan Province. This program is done once a semester, i.e., twice a year. In the first 3 years, the total number of elementary and junior high school students to be examined in each semester was approx. 2.7 millions. From August 1993, the total number increased to 3.1 millions because the senior high and senior vocational school students were added. The procedures can roughly be divided into five parts: The first part is first urinary screening. The second part is the second urinary screening. The third part is so called the third examination namely serological examination. Life guidance is introduced in the fourth part. The last part is the follow-up system. All the procedures and details will be discussed later. PMID- 11431857 TI - Mass urinary screening and follow-up for school children in Taiwan Province. AB - Mass urinary screening has been carried out among the students of public and private elementary and junior high schools in the Province of Taiwan each semester since 1990. About 3 million students were screened each time. The students who had abnormal urine screening results at the first time received a second urine analysis 10 to 15 days later to confirm the abnormal urine analysis. The blood samples of the students with abnormal urine examination were taken and biochemistry examinations including creatinine (Cr) etc. were performed since 1992. All students with abnormal urine screening results were graded by the severity of hematuria and proteinuria, the heavy proteinuria graded as "D". Chronic renal failure (CRF) is defined as impaired renal function with the serum Cr over 1.7 mg/dl. Longitudinal continuous blood and urine examinations were performed each semester for the students of grade "D" and with CRF. CRF was confirmed by either the hospital medical records or telephone visit. The purpose of this study was to delineate the prevalence of heavy proteinuria (grade D) and CRF in the students of elementary and junior high school in the Taiwan Province from 1992 to 1996. The results revealed the number of urinary screening was 10,288,620. There were 5980 cases with heavy proteinuria with four-year prevalence of 5.81 x 10(-4), 4.83 x 10(-4) for boys; 6.87 x 10(-4) for girls. Girls were affected more often than boys. The peak age of girls was 12 years old and boys was 13 years old. The number of CRF cases was 189 with the four-year prevalence of 1.84 x 10(-5), 2.24 x 10(-5) for boys; 1.41 x 10(-5) for girls. The incidence rate increased after the age of 10; the peak age of boys being 14-year old and of girls 12-year-old. The exact contributing factors, such as location on islet or lack of pediatric nephrologist, need further study. In conclusion, the four-year prevalence of heavy proteinuria in the students of the elementary and junior high schools in Taiwan was higher in girls than in boys. Glomerular nephritis (GN) is still one of the major causes of urinary abnormalities. The most-important secondary GN was systemic lupus erythematosus (SLE) with lupus nephritis. The percentage of SLE patients among anti-nuclear antibody (ANA) positive was 72%. In contrast, the four-year prevalence of CRF disease was higher in boys with the peak age at 14-year-old. GN is still the major cause of urinary screening abnormality. ANA study is indicated in all Chinese students with abnormal urinary screening. PMID- 11431858 TI - Comparison of conventional phototherapy and fiberoptic phototherapy in the very low-body-weight infants. AB - Phototherapy is effectively employed for the treatment of neonatal hyperbilirubinemia, but it may influence the physiological hemodynamics of the infants, such as skin blood flow, insensible water loss and the redistribution of cardiac output. This is a retrospective chart review study involved totally 42 very low birth weight prematures(birth body weight less than 1500 gm, VLBW) who received conventional or fiberoptic phototherapy. Infants with congenital malformation, severe cardiorespiratory instability and the requirement of diuretics, inotropics or blood transfusion were excluded. The body weight loss, daily intake/output and bilirubin level during the initial four days of phototherapy were analyzed. The results showed the efficacy of both methods was the same, while the fiberoptic therapy group had significantly more urine output(p < 0.05), the body weight loss was not statistically significant. It is possible that the fiberoptic therapy group had less insensible water loss and less attenuation of the decrease of renal blood flow, this resulted in more urine output. PMID- 11431859 TI - Pitfalls of PCR-based genotyping in patients with 21-hydroxylase deficiency. AB - Mutation analysis of CYP21A2 gene was performed in seven patients with congenital adrenal hyperplasia (CAH) by combining differential long template polymerase chain reaction (PCR) amplification and amplified created restriction site (ACRS) methods. All mutations were identified, including five alleles of deletions, three alleles of splicing (IVS2-12[C/A] > G), four alleles of Ile172Asn, and two alleles of Arg356Trp. During the course of genetic analysis of CYP21A2, we found that misgenotyping of CAH by PCR-based method is possible if both alleles of a CAH patient were deletion mutations and at least one of them carried a CYP21A1P CYP21A2 fusion gene. We also found a patient's mother was misgenotyping as IVS2 12[C/A] > G homozygous due to "allele dropout" in the PCR amplification process. We present in this article evidences of mis-genotyping by PCR-based amplification method. Due to the pitfalls observed in this study, we recommend that more methods, including microsatellite linkage analysis and direct sequencing, should be performed with direct amplification of known mutations in prenatal diagnosis of CAH to avoid misdiagnosis. PMID- 11431860 TI - Reproductive outcomes and infant health in adolescent pregnancy in Taichung city. AB - The study aimed at determining the reproductive outcomes, caregiving condition, and children's growth and development compared to those born to adult mothers. All adolescent mothers ages less than 20 years and their first born infants matched with adults mothers aged 20-29 years and their first born infants at the same neighborhoods in Taichung city in 1997 were included in this study. The results showed that there is no significant difference between children of adolescent mothers and of adult mothers in terms of the prevalence of prematurity and congenital diseases, but the children of teenage mothers have a significantly higher prevalence of low birth weight (adjusted odds ratio = 1.61, 95% CI = 1.21 2.80) than those of compared mothers. Multiple logistic regression was used to find out the risk factors for low birth weight, and found that maternal weight gain and hospitalization or not during pregnancies in mothers are significant risk factors for low birth weight. We can not find difference between the two groups in caregiving condition, such as breast feeding duration, accidental injury, diseases necessitating admission, and vaccination rates. Also there is no difference in the rate of failure to thrive or delayed developmental milestones. A long-term follow-up study on the prognosis for the adolescent mothers and their children is recommended. PMID- 11431861 TI - Recurrence of intussusception in childhood. AB - Intussusception is the most common cause of intestinal obstruction between 3 months and 6 years of age. Recurrence after reduction of intussusception in childhood is not rare. To assess the incidence and determinants of recurrence of intussusception in childhood, we conducted a prospective observation in an emergency service of a large referral center during a four-year period. We encountered 89 cases with intussusception of whom nine cases (10.1%) had episodes of recurrent intussusception. Five patients had a single recurrence, three had double recurrence and one had triple recurrence. Age of first intussusception, sex, or concurrent adenovirus infection was not related to the recurrence. None of the 27 patients who needed operative reduction had recurrence, while 9 of 62 patients who were reduced successfully by barium enema developed recurrence (P = 0.05). Compared with the first episode, significantly less vomiting, rectal bleeding and shorter duration of abdominal pain or irritable crying were noted during recurrent episodes. All the recurrent episodes were reduced successfully by barium enema. We conclude that recurrent intussusception in childhood tends to be diagnosed earlier than previous episodes and treated successfully by hydrostatic reduction without complication. Surgical reduction of recurrent intussusception may be reserved for cases of failure of hydrostatic reduction, positive peritoneal sign or existence of pathological lead point because of favorable response to barium reduction. Recurrent intussusception seldom occurs in patients who underwent surgical reduction. PMID- 11431862 TI - Lung abscess in congenital cystic adenomatoid malformation: report of one case. AB - Congenital cystic adenomatoid malformation (CCAM) of the lung is an uncommon anomaly of fetal development of terminal respiratory structures. Patients with CCAM may present either in the newborn with progressive respiratory distress, or in older children with recurrent pulmonary infections. Lung abscess is an extremely rare presentation. Herein we report an 8-year-old girl with type 1 CCAM who presented as lung abscess. PMID- 11431863 TI - Isolated fetal and neonatal ascites: report of two cases. AB - Neonatal ascites is an uncommon problem that may be caused by a number of etiologies including diseases of genitourinary system and gastrointestinal system, cardiac disease, hepatic disease, systemic infection such as TORCH or parvovirus, chylous, ovarian cause, inborn error of metabolism and idiopathic. We reported two cases of neonatal ascites, one was caused by cytomegalovirus and no obvious causes could be detected in the second one. The ascites were diagnosed by prenatal ultrasound at the gestational age of 25 weeks and 37 weeks respectively and were resolved spontaneously after birth. One-year follow-up of both cases revealed normal growth and development. No recurrent ascites could be detected by abdominal sonography except for evidence of mild hepatomegaly that was noted in case 1. Thus, isolated fetal and neonatal ascites without other concomitant abnormalities were diagnosed, a separate entity from non-immune hydrops fetalis with excellent prognosis. PMID- 11431864 TI - Steroid-induced acute psychosis in a child with asthma: report of one case. AB - A 5-year-old girl was admitted due to severe asthmatic attack. She was treated with methylprednisolone (40 mg i.v. q6h), aminophylline (loading with 5 mg/kg and maintained with 0.6 mg/kg/hr i.v. drip), nebulized terbutaline sulphate (5 mg q6h), oral procaterol 12.5 micrograms bid, along with oxygen therapy. Acute psychotic reaction with visual hallucination, delusion, panic reaction and myoclonic movement of hands developed on day 3 of admission. The patient had no previous history of psychiatric problems. The theophylline level was 9.89 micrograms/ml at the moment of psychotic reaction. After the dose of methylprednisolone was reduced from 40 mg to 20 mg i.v. q6h and shifted to other anti-asthma treatment by procaterol metered dose inhaler via spacer, the psychotic reaction disappeared a few hours later. The psychotic reaction was thought to be due to steroid therapy since no other causes could explain the psychotic reaction. PMID- 11431865 TI - Left paraduodenal hernia presenting as intestinal obstruction: report of one case. AB - We report a case of internal hernia of the small intestine in a 13-year-old boy with presentation of partial intestinal obstruction. The patient suffered from recurrent abdominal pain and chronic constipation over the past few years. An abdominal mass was suspected from clinical manifestations and images derived from abdominal echography. Upper gastrointestinal contrast study revealed poor motility at the distal jejunum with barium stasis. Follow-up film on the next day delineated medially and downwardly displaced splenic flexure and proximal descending colon. At operation, total herniation of small intestine into a retroperitoneal space through a defect on left mesocolon was noted. A left paraduodenal (mesocolic) hernia was diagnosed. The patient made an uneventful recovery after the hernia was repaired. This report provides unusual image clues of internal hernias of the small intestine presenting as ileus. Though rare, paraduodenal hernia should be taken into account in a differential diagnosis of intestinal obstruction. Early surgical intervention allows uneventful recovery to occur and also prevents the possible complication of gangrenous bowels. PMID- 11431866 TI - [Japanese-English technical terms on geriatric medicine]. PMID- 11431867 TI - [Aortic diseases in elderly patients: management of abdominal aortic aneurysm]. PMID- 11431868 TI - [Molecular medicine of hearing loss]. PMID- 11431869 TI - [Infective endocarditis in the elderly]. AB - To elucidate clinical features of infective endocarditis in the elderly, 20 elderly patients aged > or = 60 years were compared in detail with 30 others aged < 60 years retrospectively. Twelve of the 20 elderly patients had a calcific aortic valve or an artificial device as a predisposing heart disease, whereas 16 middle-aged patients had mitral valve prolapse or congenital heart disease (p = 0.001). The prevalence of major extracardiac disorders such as neurological disease were higher in the elderly than in the middle (9/20 vs 3/30; p < 0.01). The frequency of infected valve was similar; mitral in 8, aortic in 11 and other valves or congenital defect in 2 in the elderly versus 14, 11 and 6, respectively in the middle. Among 39 patients in whom causative microorganisms were identified, staphylococcus epidermidis was most frequently identified in the elderly (5/20), whereas streptococcus species was found in the middle (12/30). Time from the onset of symptoms to correct diagnosis was usually delayed in the entire group; the delay was longer particularly in the elderly than in the middle aged patients (72 +/- 87 vs 36 +/- 32 days; p < 0.1). Maximal body temperature was less in the elderly than in the middle-aged patients (38.5 +/- 0.7 vs 39.3 +/ 1.1 degrees C; p < 0.01), whereas peak level of C-reactive protein (10.4 +/- 6.1 vs 13.0 +/- 7.9 mg/dL), the incidences of heart failure (9/20 vs 10/30), and embolic complications (7/20 vs 10/20) were similar in the 2 groups. Cardiac operation was performed less in the elderly than in the middle-aged patients (9/20 vs 21/30; p < 0.08). Five elderly patients had disease-related mortality, whereas only one middle-aged patient died (p = 0.02). These results suggest that although predisposing heart disease and causative microorganism in infective endocarditis are different between the elderly and middle-aged patients, the incidence of major complications are similar. However, due to the delay of correct diagnosis in the elderly who usually have major extracardiac disorders, the prognosis of infective endocarditis in the elderly is poor. PMID- 11431870 TI - [Novel therapeutics in cardiovascular disease]. PMID- 11431871 TI - [Oral hypoglycemic agents in the treatment of elderly diabetics]. PMID- 11431872 TI - [Research progress of Alzheimer's disease-biological marker]. PMID- 11431873 TI - [Current status, clinical characteristics and treatment of alcohol-related problems in the elderly]. PMID- 11431874 TI - [Treatment and prevention of atherosclerosis]. PMID- 11431875 TI - [Learning from carotid artery]. PMID- 11431876 TI - [Comprehensive medical treatment for elderly COPD: perspective of comprehensive pulmonary rehabilitation]. PMID- 11431877 TI - [The current status of surgical treatment for COPD]. PMID- 11431878 TI - [Strategies for prevention of aspiration pneumonia]. PMID- 11431879 TI - [Clinical analysis of community-acquired pneumonia and pulmonary tuberculosis in the elderly and advances in treatment]. AB - We treated 510 elderly case (over 65 years old) among 1,017 patients with community-acquired pneumonia and 60 similar cases among 112 patients with pulmonary tuberculosis in Kawasaki Medical School Kawasaki Hospital during approximately the past 15 years. These were compared with non-elderly cases (below 65 years old). In the elderly cases with community-acquired pneumonia, atypical clinical symptoms or physical signs were frequent and the mortality rate was high because of severe underlying diseases, and poor general and nutritional conditions. Regarding a prospective study of 84 elderly cases with community acquired pneumonia during the past two years, S. pneumoniae, Respiratory virus, Gram-negative bacilli, H. influenzae, M. Tuberculosis were frequently isolated. In addition, mixed viral and bacterial infections, which were frequently noted during the winter, were significantly related to the increased frequency of community-acquired pneumonia. In treating elderly cases with community-acquired pneumonia, immunization therapy (e.g., influenza vaccine), second cephalosporin and/or macrolide antimicrobial agents for outpatients with mild pneumonia, and carbapenem and/or macrolide antimicrobial agents for hospitalized patients with moderate or severe pneumonia were most effective. The number of elderly cases with pulmonary tuberculosis has recently increased and the recognition of 10 cases was delayed because of a low percentage of positive smears, but no resistance to antituberculosis drugs have been observed. Regarding the treatment of pulmonary tuberculosis, fluoroquinolone and rifamycin derivative antibiotics have been developed as antituberculosis drugs with strong antituberculous activity. However, due to the high percentage of adverse effects in elderly patients, careful treatment with desensitization therapy for antituberculosis drugs is considered important. PMID- 11431880 TI - [Treatment for elderly lung cancer patients by anticancer drugs and possible use of target based drugs]. PMID- 11431881 TI - [New surgical treatment for lung cancer of aged patients: a prospective study for small lung cancer]. PMID- 11431882 TI - [The present and: future monitors for the care]. PMID- 11431883 TI - [Support system of activities of daily living for the elderly]. AB - People with severe disabilities use many assistive devices, such as computers and robots, to improve their quality of life. However, the elderly use only limited assistive devices, such as canes, wheelchairs, walkers, or handrails. This state is caused by two characteristics of the elderly: first, the elderly generally have difficulties in learning the operation of new equipment, especially equipment with digital interfaces. Secondly, the elderly have limited desire to improve their lives if it takes considerable effort. To solve these problems, speech recognition, gesture recognition and power assist technology have been proposed for various useful devices. Virtual reality technology and pet robots may encourage the elderly to use assistive devices to improve the quality of their lives. PMID- 11431884 TI - [The study of house monitoring system in Welfare Techno-House Mizusawa]. AB - In 1999, 8 elderly people aged over 70 stayed for 5 days in Mizusawa Welfare Techno-house and their health conditions and activities of daily life were monitored using 16 sensors attached around the house. To determine a few feasible, practical sensors and optimal sensor positions, accumulated data was analyzed. Because of bud get conditions only two sensors (infrared sensor and life line monitor sensor) were placed in one voluntary house and the data were transferred from the voluntary house to the techno-house through the ISDN and CATV lines. The data were analyzed every days. Only 2 sensors to detect were placed and attached this year, however one more sensor to detect physical conditions such as ECG records during sleeping time, will be used next year. On the other hand, an integrate system that can analyze many data transferred from various sensors simultaneously was developed. To introduce this system into privatehouse, many problems such as privacy protection, security, etc must be solved. PMID- 11431885 TI - [The current and prospective situation for the care-aids in the nursing home]. PMID- 11431886 TI - [Predictors for mortality of the community elderly--longitudinal study by TMIG LISA]. PMID- 11431887 TI - [Active life expectancy: factors influencing regional and gender differences]. AB - Active life expectancy (ALE) is defined as an expected duration to be spent with a certain level of physical/mental function. The objectives of this article are to indicate ALE values based on our prospective observation, and to discuss factors influencing regional and gender differences in ALE values. We estimated ALE without disability in basic activities of daily living (ADL) on a 5% random sample (n = 3,459) of the residents aged 65 years and over in Sendai City between 1988 and 1991. At the age 65, ALE was 14.7 years for men and 17.7 years for women. ALE occupied 91% of the total life expectancy for men and 87% for women. As compared with the reports for the American elderly, ALE was longer in Sendai than in the United States. The duration to be spent with disability was shorter among the subjects in Sendai. We estimated ALEs in three functional areas: basic ADL, instrumental ADL, and mobility, on all the residents aged 65 years and over (n = 3,590) at Wakuya Town between 1994 and 1996. For both sexes, ALE in IADL was shorter than those in basic ADL and mobility. The development and progression of disability were different between sexes: men experienced disability at a younger age and progressed at a faster rate than women. PMID- 11431888 TI - [A period between life expectancy at any age and active life expectancy]. PMID- 11431889 TI - [Evaluation and improvement of medical and nursing service and caregiving for the elderly using MDS. 3. Revised recording system of diagnoses and symptoms suitable for use in Japan]. AB - Diagnoses and symptoms are recorded by physicians in the chapter E of the Minimum Data Set (MDS), if they are relevant to disabilities of activities of daily living, cognition, behavior, medical treatments or risk of death. We improved the chapter so that it is suited to disease patterns in Japan in a format useful not only for nursing and ADL care but for medical treatment in our practice. In E1, diseases directly underlying the current disability states were recorded in the international classification of Disease, 9th Revision (ICD9). In 24, 670, 195, and 45 patients respectively, there were 0, 1, 2, and 3 separate recorded diseases. A Total of 63 ICD9 codes were observed, but only four codes; 290, 332, 431, and 434, were underlying diseases for 3% or more patients. These codes included mostly ischemic and degenerative disease of the brain. PMID- 11431890 TI - [The effect of celiprolol hydrochloride for lipid metabolism--especially for the low density lipoprotein particle size]. AB - Recently, much attention has been paid to small sized low density lipoprotein (LDL) as a risk factor for ischemic heart disease. We investigated the effect of celiprolol hydrochloride (CH), which is a beta 1 selective beta-blocker with high intrinsic sympathomimetic activity (ISA), on the LDL particle size. We treated 41 hypertensive patients with CH and studied the change in LDL particle size according to the score of fast beta lipoprotein and LDL relative mobility value (LDL-Rm) measured by lipoprotein polyacrylamide gel disc electrophoresis (PAGE). We also studied changes in blood pressure, total cholesterol (TC), trygiyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and midband on PAGE. Systolic and dyastolic blood pressure and pulse significantly decreased during treatment. TC levels were significantly decreased at 8 weeks in all subjects and at 4, 8 and 12 weeks in patients with a TC value of over 220 mg/dl. TG levels were significantly decreased at 4 and 8 weeks in patients with initial levels of over 150 mg/dl, and significantly increased at 4 and 8 weeks in those with initial levels of under 150 mg/dl of TG. HDL-C levels did not significantly change during treatment. LDL-C levels were significantly decreased at 4, 8 and 12 weeks in patients with initial levels of over 150 mg/dl. Apo AI, AII, B, CII, CIII and E levels did not significantly change during treatment. Fast beta lipoprotein scores did not significantly change overall during treatment, but were significantly decreased at 4 and 8 weeks in patients initial TG levels of over 150 mg/dl and at 4 and 12 weeks in those with initial levels of over 220 mg/dl of TC. LDL-Rm scores did not significantly change during treatment. Midband scores were significantly reduced overall at 8 weeks, and after 4 and 8 weeks in patients with initial TG levels of over 150 mg/dl and at 4, 8 and 12 weeks in those with initial TC levels of over 220 mg/dl. These results indicated that CH did not change LDL particle size. It was suggested that CH might be a beneficial beta-blocker from the standpoint of prevention for atherosclerosis. PMID- 11431891 TI - [Effect of background factors concerning both elderly persons and their families with regard to the caregiver burnout in the home]. AB - A cross sectional survey was performed to quantify factors that exhaust caregivers. We report that the degree of difficulty for caregivers correlated well with the burnout score (r = -0.517; p < 0.001), but the correlation between caregivers' burnout score and the level of their patient's basic ADL was lower (r = -0.317; p = 0.014). In this paper, we investigated other factors related to exhaustion. Caregivers' burnout score correlated with their age. The level of disease that caregivers complained increased burnout score. The need for nocturnal care and continuous observation, as well as rejection of aid burned out caregivers. Multiple regression analysis clarified that significant independent contributing factors for burnout score were help with toilet use, nocturnal aid and diseases suffered by caregivers. PMID- 11431892 TI - [Restoration of the habit of routine tooth brushing teeth in patients with severe dementia]. AB - Purpose-related activities are often lost in severe dementia, including oral hygiene habits such as brushing teeth. The aim of this study is to investigate whether patients with severe dementia can be induced to develop an oral hygiene routine through an occupational therapy approach and whether routine training in oral hygiene habits can reduce dental or denture plaque. Six female inpatients in the same room, who had been given diagnosis of multiple cerebral infarction and dementia. The project design included 4 periods a, b1, b2, and c, a and c were observation periods, while b1 and b2 were general approach periods. In the approach periods we held meetings, and made the program for oral hygiene, planning and problem-solving. In the b2 period both direct intervention by the dentists, dental hygienists and occupational therapists reinforced the habit of brushing teeth in subjects and educated care approach guidance. No patient completely resumed routine habit of brushing teeth, but an improvement in brushings was found in four subjects, one case had no change, and in one case brushing deteriorated. The state of oral hygiene showed improvement in periods b1 and b2. Although the adherence of dental plaque decreased in period b2, it increased in period c. Despite the improvement in performance, sufficient improvement in oral disease prophylaxis was not recognized, suggesting the necessity of professional oral health care by dental staff, and of a disciplinary team approach. PMID- 11431893 TI - [Effects of aging on oxidisability of low density lipoprotein]. AB - Oxidised LDL is taken up by macrophages via scavenger receptors, leading to foam cell formation and is thus considered to contribute to atherogenesis. Aging results in the increase of lipids and the decrease of antioxidant enzyme activity in serum. In this study, we investigated the effects of aging on LDL oxidisability. We measured LDL oxidation lag time, plasma lipids, albumin and uric acid were examined in 306 Japanese (169 men, 137 women). The mean +/- SE of LDL oxidation-lag time in subjects was 58.9 +/- 1.0 min. The lag time (80.3 +/- 4.8 min) was longest in subjects in their 20 s and shortest in those in their 40 s (58.9 +/- 1.0 min). The longest lag time was in second-decade men (88.9 +/- 6.2 min) and shortest in fourth-decade women (50.7 +/- 2.2 min), and these results were similar even excluding subjects with abnormal biochemical data (total cholesterol, triglyceride, GOT, GPT, gamma GTP, creatinine and glucose). We analyzed the effects of various factors on lag time using multiple linear regression. Aging, uric acid and LDL-cholesterol significantly influenced lag time. Our results suggest that LDL oxidisability might been regulated by aging, changes in LDL-cholesterol with aging and variations in physical antioxidant function. PMID- 11431894 TI - [Questionnaire survey comparing first-year medical students and non-medical students with regard to their level of interest and knowledge of care and terminal care for the elderly]. AB - To study the level of interest and knowledge about care and terminal care for the elderly of medical students in Nagoya University, we made a questionnaire for first-year medical students (about 90 students per year) each April from 1998 to 2000. To assess the differences, we also gave the same questionnaire to 75 first year non-medical students in 1999. Only about 10% of students had experienced providing care for member(s) of their family, but about one third stated that other family member(s) such as their mother, cared for older family member(s). The introduction of long-term care insurance was known by 22%, 43.5% and 96.6% of medical students in 1998, 1999 and 2000, respectively. Moreover, 19.8%, 13.0% and 52.3% of medical students knew about the care managers, respectively. About 80% of students hoped to be informed about their disease if they had terminal-stage cancer. However, about half of students could not decide whether to tell a family member who had terminal-stage cancer. Medical students had significantly more interest in the care system (68.5% in 1999) and terminal care (72.8% in 1999) for the elderly than non-medical students (41.3% and 40.0% in 1999, respectively). There were no significant differences between medical and non-medical students with regard to experiences of care, knowledge of long-term care insurance system and the attitudes to informing about terminal-stage cancer. However, more medical students had an interest in the elderly care system and terminal care than non medical students. PMID- 11431895 TI - [Quantitative evaluation of the burden of those giving home care for senile dementia of Alzheimer type subjects using the burnout scale of pines]. AB - We quantitatively measured the physical and psychological burden of caregivers of 25 patients with senile dementia of Alzheimer type (SDAT). The Barthel Index (BADL, full score: 20 points) and the caregiver burden in terms of physical symptoms correlated well (r = -0.964, p < 0.001), as did the degree of abnormal behavior and caregiver burden in terms of psychological symptoms (r = 0.946, p < 0.001). The correlation with the burnout scale (BOS) of Pines was best when both factors of psychological and physical symptoms were included. The correlation between BOS and the caregiver burden in terms of both physical and psychological symptoms was r = 0.874, p < 0.001, and the correlation between BOS and "the degree of abnormal behavior" +(20- "BADL") was r = 0.853, p < 0.001. The burden in terms of physical symptoms increased as the BADL score decreased, but the burden in terms of psychological symptoms increased initially and decreased in the last phase of the disease. We conclude that the BOS score of SDAT caregivers was stable in the initial phase, then increased rapidly, thereafter preserved high, and dropped rapidly as the BADL score decreased. PMID- 11431896 TI - [Cognitive function and brain atrophy in elderly type 2 diabetic patients in comparison with non-diabetic elderly subjects]. AB - Previous studies have suggested that type 2 diabetic mellitus could lead to learning and memory deficits. We studied cognitive function tests and brain computed tomography (CT) findings in elderly subjects with drug-treated type 2 diabetic patients (n = 9), diet-treated type 2 diabetic patients (n = 8) and nondiabetic subjects (CR, n = 21). A battery of cognitive function tests (Cog-T; WAIS-R's digit span test and symbol test, Stroop Test, ADAS's verbal memory test, and MMSE) was carried out on two occasions, separated by at least 6 months. Brain CT was analyzed by the following 5 variables; 1) Evan's Ratio, 2) Inverse Cella Media Index, 3) maximum width of the third ventricle, 4) maximum width of temporal horn tips on both sides and 5) maximum width of the Sylvian fissure at the insula, bilaterally. The scores of Cog-T did not differ significantly between the groups. On brain CT measurements, maximum width of the temporal horn tips on right side were significantly different in the three groups (ANOVA, P = 0.035). The drug-treated diabetics subjects had wider temporal horn tips on the right side than did the diet treated diabetics and nondiabetic subjects (Fisher's post hoc test, P = 0.030, P = 0.016). PMID- 11431897 TI - [Evaluation of the mental state of elderly diabetic patients in education]. AB - Maintaining to good glycemic control is necessary to treat diabetes effectively and it is also necessary to understand the mental characteristics of elderly patients with diabetes. We investigated evaluation of the mental state, using self-depressive scale (SDS) and the Tokyo university egogram form (TEG), of 356 asymptomatic diabetic patients (254 non-elderly, 102 elderly) admitted for educational about diabetes. On TEG, we studied evaluation of five ego states for the following showing: critical parent (CP), nurturing parent (NP), adult (A), free child (FC) and adapted child (AC). First of all, we studied evaluation of mental characteristics of the elderly, NP and A of TEG were higher in the elderly than in the non-elderly (p < 0.01). Furthermore, the SDS was lower on discharge than on admission in both groups (p < 0.01). The group showing improvement had a significantly higher SDS score on admission than the group showing aggravation (p < 0.01). We classified the group with an SDS score 50 or more SDS as the depressed group; those with a 40 to 49 score as the slight distress group; and a score of 39 or less score was considered normal. The depressed group had significantly lower A and FC, and a higher AC on TEG, than other groups (p < 0.01). Therefore, elderly diabetic patients seem to be rational and obliging. This study revealed the mental effects of diabetic education to be similar in elderly and non-elderly diabetic patients. PMID- 11431898 TI - [Factors contributing to dying at home in elderly patients who received home care service]. AB - This study examine whether around-the-clock medical support is a contributing factor to dying at home, and also tried to identify other such factors. Visiting nursing records of 81 elderly patients who died at home or in hospital after receiving home care at two facilities, one with and one without 24-hour medical support respectively were examined retrospectively. The subjects were divided into two groups: those who died at home and those who died in a hospital or nursing home. The two groups were compared in terms of clinical and sociodemographic characteristics and preferences for dying at home, expressed by patients, families and medical staff. Those who died at home showed a significantly higher rate of total dependence (84.6% vs. 48.1%) at month before death. Dying at home was significantly more preferred by all patients, families and medical staff. The major reason for hospitalization was rapid deterioration of the patients' condition. The facility with around-the-clock medical service had a higher rate of dying at home (42% vs. 27%, p = 0.18). Also, patients, families, and medical staff associated with their facility showed a higher preference for dying at home. We concluded from the above that the contributing factors for dying at home are: 1) total dependence level of ADL at one month before the death, and 2) preference for dying at home expressed by the patient, family and medical staff. This study suggests 24-hour medical support should be a requirement for in-home terminal care. Supporting advice from the staff to the family seems to be another contributing factor. PMID- 11431899 TI - [Low dose melphalan therapy was effective in an elderly patient with MDS-AML]. AB - An 80-year-old man with MDS-refractory anemia (RA) suffered transformation to a leukemic state after 18 months. The karyotype of the bone marrow cells was 47, XY, +8 in 8 cells among 20 dividing cells analyzed. Combination therapy of 150 micrograms of granulocyte colony-stimulating factor (G-CSF) and 250 mg of cytarabine ocfosfate (SPAC) for 3 weeks had no beneficial effect. Then, the patient was subjected to low-dose (2 mg daily) melphalan therapy. Gradual and concurrent improvement in anemia, thrombocytopenia, and neutropenia occurred, and the patient became free of transfusions at 2 weeks after the treatment began. Since then, his performance status has improved from grade 4 on his diagnosis of AML to grade 2. Cytogenetic analysis was normal in all 20 dividing cells in the bone marrow examination and melphalan had no adverse effect. Recently, several reports of low dose chemotherapy for elderly patients or high risk leukemia have been described, and have sustained for the QOL therapy. In the present case, low dose melphalan therapy was effective and, moreover the abnormal karyotype of trisomy eight had disappeared. PMID- 11431900 TI - [Noninvasive positive-pressure ventilation in an elderly woman with acute respiratory failure caused by influenza a virus pneumonia]. AB - A 79-year-old woman was admitted on January 18, 2000, with a lumbar compression fracture. Two days after admission, a chest X-ray film showed reticular infiltrates in the right lung field. She was diagnosed as having pneumonia and was treated with antibiotics. Despite this therapy, her symptoms did not improve and the infiltrates diffusely extended to involve both lungs on chest X-ray films. She was placed on noninvasive positive pressure ventilation (NIPPV) for progressive respiratory failure at 5 days after admission in order to avoid endotracheal intubation. Her hypoxemia was immediately improved by oronasal bilevel positive airway pressure ventilation (BiPAP) and chest X-ray films revealed improvement of the reticular infiltrates. Measurement of viral antibody titers showed that the cause of acute respiratory failure was influenza A virus pneumonia. In conclusion, NIPPV may reduce the need for intubation of elderly patients with acute respiratory failure, as well as chronic obstructive pulmonary disease (COPD) patients. PMID- 11431901 TI - [Weight reduction improves high blood pressure and microalbuminuria in hypertensive patients with obesity]. AB - We investigated the effect of weight reduction on blood pressure, microalbuminuria and renal function in hypertensive patients with obesity for over 12 months. Twenty-five patients with a body mass index (BMI) of over 25 were prescribed low calorie diet (25 kcal/kg). All patients had mild hypertension and microalbuminuria. They were classified into 2 groups after 12 months. Group A consisted of 10 patients who had a weight loss of at least 5%. Group B consisted of 15 patients who did not have any weight loss. The following results were obtained. (1) The percentage of patients with hyperfiltration (GFR; more than 140 ml/min) was 20%. (2) Blood pressure, fasting plasma insulin level, urinary sodium and albumin excretion rate were significantly decreased in Group A. On the other hand, these changes were not observed in Group B. (3) Reduction in mean arterial blood pressure significantly correlated with the fall in body weight. (4) Renal function did not change during the study period in both groups. (5) Urinary albumin excretion rate significantly correlated with weight reduction, decrease in blood pressure and fasting insulin levels. Blood pressure and urinary albumin excretion rate in hypertensive patients with obesity significantly decreased with weight reduction. Probably, weight loss improves insulin resistance and decrease in the plasma insulin level causes a reduction in blood pressure and urinary albumin excretion rate. PMID- 11431902 TI - [A proposed cooperation system between physicians in private practice and hospitals to prevent neglect of poor prognostic cases with mild urinary abnormalities]. AB - Some outpatients with only mild urinary abnormalities have histologically active lesions. To prevent the progression of these lesions, they should not be overlooked at the initial diagnosis. For this purpose, I propose a new system of diagnostic cooperation between physicians in private practice and hospitals. A 50 year-old male showed proteinuria([symbol: see text]), occult blood([symbol: see text]), and one granular and red cell cast in the total fields in spot urine samples, a 24 hr urinary protein of 0.17 g, a serum creatinine of 1.2 mg/dl and a mean blood pressure of 112 mmHg at the first examination. Four months after the administration of an ACE-inhibitor, the urinary protein decreased to 0.04 g while the serum creatinine level increased to 1.5 mg/dl. Therefore, a biopsy was performed at Kyoto University Hospital. Twenty-one glomeruli were obtained revealing 6 with global sclerosis and 13 proliferative glomeruli, including 3 with crescentic and 4 with adhesive lesions. In addition, interstitial cell infiltration was observed. After treatment for active blomeruli lesions using prednisolone and warfarin, the 24 hr urinary protein decreased to 0 g and serum creatinine to 1.2 mg/dl, Since a correlation between the relative volume of the renal cortical interstitial involvement and the serum creatinine concentration has been reported, the possibility of glomerular-interstitial interaction was considered. Thus, to prevent the neglect of patients with apparently mild disease associated with a poor prognosis, I propose the establishment of a system of diagnostic cooperation consisting of physicians in private practice, general hospitals and central hospitals that can provide appropriate treatment principles and give instructions for patients with mild urinary abnormalities encountered in daily practice. PMID- 11431903 TI - [A case of myeloma kidney complicated by extramedullary plasmacytoma with massive bleeding]. AB - A 76-year-old woman, who had received hemodialysis due to chronic renal failure of unknown cause for two months, was admitted to our hospital. She was suffering from severe pain in the left thigh, rapidly progressive anemia and thrombocytopenia after receiving a contusion on her left thigh. Soon after admission, the patient died of shock. Autopsy revealed multiple myeloma(lamda type) with extramedullary plasmacytoma and systemic amyloidosis. In the kidney, there were typical tubular casts with multinucleated giant cells and interstitial fibrosis. More specific findings included an extramedullary plasmacytoma in the left iliopsoas muscle surrounded by a huge hematoma. Internal hemorrhage resulting from indirect contusion at this site was likely to have caused her shock. Since typical clinical findings of multiple myeloma, such as serum M protein and hypercalcemia, were not found in this case, it was difficult to make a diagnosis of multiple myeloma. In case of multiple myeloma, micro- or macroscopic extramedullary tumor formation is not rare, but there has been no report of a case with macroscopic tumor formed in skeletal muscle, exhibiting massive hemorrhage. We report here a case of multiple myeloma with an unusual clinical course. PMID- 11431904 TI - [A case of silicosis with MPO-ANCA-associated glomerulonephritis and alveolar hemorrhage]. AB - A 63-year-old man was referred to our hospital for rapid deterioration of his renal function. He had worked as a metal founder for more than 40 years, and had been diagnosed as having silicosis. Laboratory data on admission showed severe anemia, thrombocytopenia, and end-stage renal failure (BUN 88.8 mg/dl, serum creatinine 9.0 mg/dl). Myeloperoxidase anti-neutrophil cytoplasmic antibody(MPO ANCA) was also detected in his sera. On the next day after admission, he complained of sudden dyspnea and hemoptysis. Mechanical ventilation with pure oxygen was insufficient to improve hypoxia without concomitant use of percutaneous cardio-pulmonary support(PCPS) and continuous hemofiltration(CHF). We diagnosed his condition as MPO-ANCA-associated rapidly progressive glomerulonephritis with diffuse alveolar hemorrhage. Treatment with plasmapheresis, pulse methylprednisolone and pulse cyclophosphamide effectively improved his hemoptysis as well as chest X-ray findings and blood gas analysis. However on his later clinical course, he was complicated with superimposed complex infection and passed away. Autopsy findings showed crescentic glomerulonephritis in the kidneys and silica nodules in the lungs. Recently it has been postulated that some relationship exists between ANCA associated(especially MPO-ANCA-associated) glomerulonephritis and silica exposure. The reported cases of glomerulonephritis in the patients with silica exposure showed a rapidly progressive clinical course and pauci-immune necrotizing crescentic glomerulonephritis in their histology. Gregorini et al. reported that 12 of 37 (32%) male patients with RPGN had either silicosis or significant silica exposure, and 7 of 8 patients examined were ANCA-positive(6 of 7 were MPO-ANCA-positive). Therefore silica seems to cause glomerulonephritis by disrupting the immune response. Including this case mentioned above, we have experienced 10 cases of MPO-ANCA-associated glomerulonephritis, at least 3 cases out of which had suffered from silicosis in the past(30%). These results indicate that silicosis should be considered a relevant pathogen of MPO-ANCA-associated glomerulonephritis beyond the race. PMID- 11431905 TI - [A case of acute renal failure and liver dysfunction induced by carbamazepine(CBZ)]. AB - A 56-year-old female with symptomatic epilepsy was admitted to our hospital because of acute renal failure(ARF) and liver dysfunction(LD) after receiving CBZ for two months. She had suffered a drug eruption caused by phenobarbital and valproate six months previously. Renal and liver biopsies presented acute interstitial nephritis and active chronic hepatitis, respectively. Drug-induced lymphocyte stimulating test showed CBZ positivity. Steroid therapy resulted in recovery from ARF and LD. CBZ sometimes causes ARF or LD, but rarely induces both simultaneously, especially in adults. Pathological evidence of two lesions other than from autopsy seems to be the first step in this case. Cross reaction with other antiepileptic agents was also of interest, suggesting that one member of the cytochrome P450 subfamily, CYP3A, participated in the mechanism. PMID- 11431906 TI - [Tubulointerstitial nephritis in the case of acute renal failure from sepsis after a cat bite]. AB - Acute tubulointerstitial nephritis is associated with a variety of causes, such as drug interaction, and infectious or immunological mechanisms. We describe a patient who suffered from sepsis, septic shock, disseminated intravascular coagulation(DIC), hepatic failure and renal failure after receiving a bite from her house cat. The causes of her acute renal failure were initially thought to be due to circulatory failure with hypotensive shock, decrease in renal blood flow with fibrin formation by DIC, or microangiopathy such as hemolitic uremic syndrome. However, the renal biopsy on the 60th hospital day indicated tubulointerstitial nephritis, which was recognized by the presence of patchy and focal mononuclear small cell infiltration with invasion to the tubular epithelium. We concluded that prolonged renal failure was caused by tubulointerstitial nephritis. The cause of tubulointerstitial nephritis was not identified. Tubulointerstitial nephritis should be taken into consideration when the recovery from acute renal failure is slow. PMID- 11431907 TI - [The role of airway submucosal glands in the airway mucosal defense system]. AB - Despite constant inhalation of air-borne particles including a variety of microbes and antigens, the normal lungs hardly ever develop infection or airway injury. This is because the normal lung is equipped with sophisticated defense mechanisms against foreign substances. It has been reported that the airway mucosa, especially the submucosal glands, play important roles not only in nonspecific defense using airway secretions but also in specific defense in cooperation with immune cells. In contrast to the nasopharyngeal or intestinal mucosa, which is always exposed to many foreign antigens, the mucosal surface of the lower respiratory tract in known to be kept in a germ-free condition. This fact indicates that immunological information derived from the antigen-rich mucosa, such as the intestine, might be transmitted to the airway mucosa, thus resulting in efficient removal of unwanted substances. This immunological elimination requires specific antibodies (Abs) against harmful antigens, and the major population of Abs in the airway is dimeric IgA. Airway submucosal glands synthesize a secretory component (SC), a transporter of secretory IgA, and immunoglobulin-containing plasma cells have been identified preferentially around the glandular tissue. Overall, the submucosal glands play a key role in the integrity of airway mucosal immunity. PMID- 11431908 TI - [Opportunistic pneumonia after kidney transplantation]. AB - This study was conducted to evaluate clinical features at the onset of pneumonia and the usefulness of methods for diagnosing pneumonia in patients who have undergone kidney transplantation. From January 1990 to December 1998. 174 kidney transplantations were performed, and were followed by 22 cases of pneumonia. Of the 22 pneumonia patients, 16 were male and 6 were female. The median age of the 22 patients was 37.2 +/- 13.3 years. All the patients received cyclosporin A and corticosteroids. In 11 cases, the organisms were identified in the microbiology or pathology laboratory, either during life or at autopsy. Six cases were due to Pneumocystis carinii (PC), three to PC and Cytomegalovirus (CMV), one to Aspergillus, and one resulted from miliary tuberculosis. Pneumonia occurred within 4 months after kidney transplantation in most cases. The mean interval between the transplantation and the appearance of pneumonia was 77.3 +/- 34.3 days, except in the cases of Aspergillosis and miliary tuberculosis, where the intervals were 46 and 50 months, respectively. The mean interval from the appearance of symptoms to the detection of pulmonary infiltration was 3.3 +/- 4.3 days. The clinical features present when pulmonary infiltration was detected by CT were fever (91%), cough (32%), and crackles (27%). However, at this time, 55% of the cases had no symptoms other than fever. Chest radiographs were positive for pulmonary infiltration in 64% of the cases at the same time that the pulmonary infiltrates were detected by CT. Eighty percent of the cases exhibited diffuse interstitial infiltrates. Organisms were detected in 7 of 9 cases examined with bronchofiberscopy (BF). But in only one of 13 cases that did not undergo BF. Increased values of serum beta-D-glucan were detected in the early phase of three PC pneumonia cases, suggesting that beta-D-glucan is useful as a marker of PC pneumonia. The use of bronchofiberscopy was more frequent in survivors of PC pneumonia than in non-survivors, whereas the mean age was higher and coexisting CMV infections were identified more frequently in the non survivors. We concluded that fever is important as an initial symptom of pulmonary infection. In addition, we find that CT is very useful for the detection of interstitial infiltrates, and BF is an excellent method for detecting organisms in the pneumonia patient after kidney transplantation. PMID- 11431909 TI - [Clinical comparison of Chlamydia pneumoniae pneumonia, ornithosis, and Mycoplasma pneumoniae pneumonia]. AB - To elucidate the differences between the clinical aspects of Chlamydia pneumoniae (C. pn) pneumonia and those of two other atypical pneumonias, Chlamydia psittaci (C. ps) pneumonia and Mycoplasma pneumoniae (M. pn) pneumonia, we analyzed the symptoms and laboratory data on the cases of these three types of pneumonia: 46 cases of C. pn pneumonia, 39 cases of C. ps pneumonia, and 131 cases of M. pn pneumonia. C. pn pneumonia was significantly more frequent among the elderly (mean 70 +/- 16 years, p < 0.01) and patients were significantly more likely to be male (76%, p < 0.05). A white blood cell count of over 10,000 was seen in 46% of C. pn pneumonia cases, a higher proportion than those of C. ps pneumonia (15%, p = 0.03) or M. pn pneumonia (18%, p = 0.006) cases. The proportions of patients with these infections who had an elevated GOT or GPT were not significantly different. Maximum body temperature was higher in M. pn pneumonia than in C. pn pneumonia (p = 0.003). Purulent sputa were seen in 44% of C. pn pneumonia cases and 50% of M. pn pneumonia cases, and these rates were higher than that of 13% in C. ps pneumonia cases (p = 0.002, p = 0.004). Dyspnea and anorexia symptoms were the most frequent in C. pn pneumonia cases (24% and 29%, respectively, the highest of all three pneumonias). There were clinical differences between C. pn pneumonia and the other two atypical pneumonias. However, there was some difficulty in differentiating between C. pn pneumonia and typical bacterial pneumonia because mixed infections were common (24%) in C. pn pneumonia cases. PMID- 11431910 TI - [Dumon stent placement via endotracheal tube]. AB - We conducted a Dumon stent placement via endotracheal tube for 10 patients with airway stenosis. The conventional endotracheal tube is inserted beyond the stenosis site; this procedure is conducted with the use of a flexible bronchoscope under general anesthesia. The Dumon stent is folded and inserted into the endotracheal tube and is then introduced into the stenosis site with the use of a cylindrical-tipped stainless steel wire as a pusher. Although the Dumon stents were placed using a rigid bronchoscope for the first 7 patients, the present procedure was used for the latest 10 patients. Compared with the rigid bronchoscope technique, this procedure is suitable for the placement of a larger stent for a shorter time. It has the following advantages over the rigid bronchoscope technique: (1) the use of an endotracheal tube and flexible bronchoscope makes the stent placement easier for the practitioner and less stressful for the patient; (2) because of the flexibility of the endotracheal tube, a Dumon stent can be placed easily, even in the left main bronchus or in a markedly shifted trachea or bronchus, and also in a patient who has difficulty in expanding the neck; (3) a stent can be placed safely in a patient with severe tracheal stenosis and orthopnea. The present procedure does, however, have the disadvantage that it is difficult to control the direction of the tip of the endotracheal tube. We concluded that the present procedure could be a useful method for Dumon stent placement. PMID- 11431911 TI - [Ganglioneuroma detected in a patient over age 60]. AB - We report a case of ganglioneuroma in a 67-year-old woman who presented with an abnormal shadow at a medical examination. She was admitted and chest radiography disclosed a mass in the upper right mediastinum. We suspected a mediastinal tumor after chest CT, chest MRI and bronchofiberscopic examination, and so surgical treatment was performed. The histopathological diagnosis was ganglioneuroma. Ganglioneuroma is thought of as a children's disease and adult onset is rare. We reasoned that ganglioneuroma should be included among the mediastinal tumors in patients over 60. PMID- 11431912 TI - [A case of bronchioloalveolar carcinoma with multiple cystic shadows on chest CT]. AB - A 68-year-old woman was admitted to our hospital in 1998 with shortness of breath on exertion. On the first admission, chest radiography revealed multiple nodular shadows and areas of ground-glass appearance mainly in the basal lung fields, and chest CT scans showed multiple thin-walled cystic lesions in both lung fields. Bronchioloalveolar carcinoma was diagnosed by transbronchial lung biopsy. The CA 19-9 level in the serum was abnormally high. The disease progressed, and the patient died 7 months after diagnosis. We report this case because CT scans showed multiple cystic lesions, which is very rare. The mechanism of cystic formation in this case of lung cancer may have involved disruption of the alveolar structure and enlargement of disrupted spaces or of the check valve mechanism. PMID- 11431913 TI - [Hypersensitivity pneumonitis caused by a factory humidifier. A case report]. AB - A 64-year-old man was hospitalized for productive cough and dyspnea. Both chest radiographs and CT scans showed areas of ground-glass opacity in the middle and lower lung fields on both sides. The BAL and TBLB findings were compatible with hypersensitivity pneumonitis. The serum was negative for antibodies against Trichosporon species, and the result of a lymphocyte stimulating test for administered drugs including a Chinese medicine was also negative. A humidifier was suspected as the cause because it had been used for more than 10 years in the factory where the patient had been working. An inhalation test using the humidifier fluid successfully provoked dyspnea, fever and fine crackle, and laboratory tests demonstrated hypoxemia, reduction in vital capacity and the elevation of CRP. Agar gel diffusion using the patient's serum showed a precipitating line against Cephalosporium acremonium, but this line did not fuse with any precipitating line formed between the humidifier fluid and the serum, indicating that no Cephalosporium was Present in the humidifier fluid. Since a high level of beta-D glucan was detected in the humidifier fluid, an unidentified fungus was suspected to be the antigen. PMID- 11431914 TI - [A case of pulmonary thromboembolism with heparin-induced thrombocytopenia]. AB - In February 2000, a 70-year-old man was admitted to our hospital complaining of back pain and dyspnea on exertion. Pulmonary thromboembolism was diagnosed, and he was treated with intravenous urokinase and heparin. The pulmonary thromboembolism improved, though heparin-induced thrombocytopenia (HIT) was subsequently observed. The thrombocytopenia was then improved by withdrawing the intravenous heparin, but thrombosis appeared extending from both femoral veins to the inferior vena cava. The thrombosis was dispersed by catheter-directed thrombolysis. There have been few reports of HIT in Japan. Heparin is frequently used for the treatment of pulmonary thromboembolism, but special care must be taken, since severe thrombotic complications are associated with HIT. PMID- 11431915 TI - [Pleural effusion and eosinophilia associated with Metagonimus yokogawai infection]. AB - We encountered a patient with a mild fever and chest discomfort in whom right pleural effusion and eosinophilia (48%) had developed. Parasite eggs were found in the feces, and metagonimiasis was diagnosed. The pleural fluid contained many Charcot-Leyden crystals, suggesting active eosinophilic inflammation. Metagonimus yokogawai usually occurs as a gastrointestinal parasite and causes only mild gastrointestinal symptoms such as diarrhea. Most cases of metagonimiasis resolve spontaneously. However, the main manifestation of metagonimiasis in our patient was pleural effusion. Some cases of idiopathic eosinophilic pleural effusion of unknown origin resolve spontaneously, as in our patient. Patients who are therefore considered to have idiopathic pleural effusion may include those with transient parasitic infection. PMID- 11431916 TI - [Follicular bronchiolitis associated with pulmonary tuberculosis--report of a case]. AB - The patient, a 39-year-old man, presented in May 1997 with an untreated persistent cough with excessive sputum of 5 years' duration. He was admitted to hospital because of the severity of the symptoms and the presence of acid-fast bacilli in his sputum. These bacilli were identified as Mycobacterium tuberculosis by the polymerase chain reaction method. After treatment with antituberculous drugs was initiated, his symptoms and the patchy infiltrative shadows on his chest radiographs gradually resolved. However, the patient continued to expectorate a purulent sputum, showed diffuse micronodular shadows on chest radiographs, and had hypoxemia as well as mixed ventilatory and small airway disturbances on pulmonary function tests. We performed a video-assisted thoracoscopic biopsy of the lung, which revealed follicular bronchiolitis. PMID- 11431917 TI - [Pulmonary Nocardia otitidis-caviarum infection in a patient with Cushing's disease]. AB - While Nocardial infections are being diagnosed with increasing frequency, infection with Nocardia otitidiscaviarum remains relatively uncommon. We report a case of pulmonary Nocardia otitidis-caviarum infection in a 35-year-old man with Cushing's disease. This work describes the first case of nocardiosis in Japan caused by Nocardia otitidis-caviarum in Cushing's disease. The patient was admitted to our department because of edema. A diagnosis of Cushing's disease was made on the basis of elevated serum levels of cortisol and adrenocorticotropic hormone (ACTH) and pituitary adenoma was found in a cranial CT scan. One month after admission, chest radiographs showed a large bilateral mass on the lung fields. Nocardia otitidis-caviarum was isolated from the sputum. The patient responded poorly to intravenous PAPM/BP, but later improved after treatment with trimethoprim-sulfamethoxazole, but he died of heart failure and respiratory failure after the initiation of this therapy. This case demonstrated that nocardiosis must be considered in differential diagnosis as an opportunistic infection. PMID- 11431918 TI - [Minocycline-induced pneumonitis presenting as multiple ring-shaped opacities on chest CT, pathologically diagnosed bronchiolitis obliterans organizing pneumonia (BOOP)]. AB - A 39-year old woman was admitted to our hospital because of cough and abnormal shadows on chest radiographs. She had been treated for 5 months for acne vulgaris with minocycline hydrochloride (MINO). Chest computed tomographic (CT) scans showed multiple ring-shaped opacities in both lungs. Bronchoalveolar lavage disclosed an increase in the total number of cells and a marked increase of lymphocytes. A lung specimen obtained by transbronchial lung biopsy (TBLB) was pathologically diagnosed as bronchiolitis obliterans organizing pneumonia (BOOP). Withdrawal of minocycline led to rapid remission without treatment. The clinical course and histological findings for TBLB suggested that this case was minocycline-induced BOOP. Several cases with minocycline-induced pneumonitis have been reported. However, there are few reported cases of minocycline-induced BOOP, the present case being only the second found in the literature. PMID- 11431919 TI - [A case of chronic hypersensitivity pneumonitis due to wild pigeons]. AB - A 61-year-old woman was admitted to the Oita Medical University Hospital because of a nonproductive cough and exertional dyspnea. Interstitial changes had been seen on her chest radiograph 5 years previously, but no respiratory symptoms were identified at that time. On admission, chest radiography revealed linear and ground-glass opacities in the middle and lower lung fields. Computed tomography provided evidence of bronchiectasis and micro-honeycombing of the lungs, while lymphocyte and neutrophil counts in the bronchoalveolar lavage fluid were increased. Transbronchial lung biopsy demonstrated alveolitis and Masson's bodies. The patient was not a pigeon breeder, but she could have been exposed to pigeons at her workplace. Indeed, she had specific antibodies against pigeon serum and droppings, and her peripheral lymphocytes showed proliferation in response to pigeon serum. A positive provocation test involving inhalation of pigeon serum confirmed that she had chronic hypersensitivity pneumonitis caused by allergy to pigeons. This is a rare case of chronic hypersensitivity pneumonitis associated with wild pigeons, that progressed to pulmonary fibrosis. Antigen provocation testing proved to be of great value. PMID- 11431920 TI - [The usefulness of MR angiography (MBA) in the diagnosis of pulmonary thromboembolism]. AB - We report a case of acute pulmonary thromboembolism (PTE) that was accurately diagnosed as PTE by magnetic resonance angiography (MRA). The patient was a 69 year-old woman who exhibited dyspnea. On chest radiography, neither infiltrative shadows nor tumor shadows were observed. Since the lung perfusion scintigram showed segmental defects in both lung fields, MRA was performed. On MRA, blockage of the artery and the perfusion defects in both upper lobes could be seen. We confirmed the diagnosis as PTE on the basis of these typical MRA findings. Our preliminary findings suggest that MRA shows promise as a safe, accurate imaging technique for the diagnosis of PTE. PMID- 11431921 TI - [Caveolins and lipids]. PMID- 11431922 TI - [Role of membrane phospholipid dynamics in spatio-temporal regulation of cell membrane structure]. PMID- 11431923 TI - [Lipid domains in the endocytic pathway]. PMID- 11431924 TI - [Glycolipid assembly/rafts and cellular functions]. PMID- 11431925 TI - [Cell-intrinsic timer regulating oligodendrocyte development]. PMID- 11431926 TI - [Generation of mice with mitochondrial dysfunction by introducing mouse mtDNA carrying a deletion into zygotes]. PMID- 11431927 TI - [Comparison with other sequences: sequence similarity searches]. PMID- 11431928 TI - [Pathogenesis of surface component from Treponema denticola]. PMID- 11431929 TI - [Chemical structures and serological specificity of lipopolysaccharides from Helicobacter pylori]. PMID- 11431930 TI - [Studies on DNA replication mechanism and related proteins in Archaea]. PMID- 11431931 TI - [Activity of bicozamycin against Escherichia coli O157:H7 producing Vero toxin]. AB - Bicozamycin (BCM) which inhibits protein synthesis by inhibiting Rho-dependent transcription termination factor ATPase activity is used for treatment of animal infections. BCM had moderate activity of MIC 16-32 micrograms/ml against enterohemorrhagic Verotoxin (VT)-producing Escherichia coli (EHEC) O157:H7 and inhibited production of VT1 and VT2. The activity of BCM against EHEC was slightly higher in anaerobic conditions, and was more evident in vivo. BCM decreased CFUs of EHEC in caecum more effectively than fosfomycin, cefixime and norfloxacin in a mouse infection model. Moreover, BCM did not increase the amount of either VT1 or VT2 in caecum in mice. In contrast, norfloxacin increased mortality of mice infected with EHEC by inducing VT production. The results suggest that BCM is useful for the treatment of EHEC infection and eradication of EHEC. Dairy live stock, especially young animals, have been implicated as a principal reservoir of EHEC. Eradication of EHEC from live stock will become an important problem in the near future. The narrow spectrum antibiotic BCM is expected to be a safe and effective antibiotic to eradicate EHEC from live stock. PMID- 11431932 TI - [Diagnostic imaging in male pelvic disorders]. AB - This article reviews the imaging findings of male pelvic disorders, especially those of prostate disease. Magnetic resonance imaging (MRI) provides excellent anatomical detail, and is useful to stage prostate cancer and to understand the process of anomaly. However, failure to detect capsular invasion remains a significant weakness in prostate cancer. It is important to understand the limits and contributions of MRI to the staging of prostate cancer, and to perform the appropriate examination. PMID- 11431933 TI - [Diagnosis of soft tissue tumors: 10 checkpoints]. AB - This review, presents 10 checkpoints for the diagnosis of soft tissue tumors and the MRI reporting method for the general radiologists. The 10 checkpoints consist of the following, (1) clinical information, (2) calcification, (3) characteristic shape, (4) multiplicity, (5) lymph node enlargement, (6) characteristic sign and appearance, (7) internal morphology (fatty component, myxoid matrix, collagenous tissue), (8) flow void, (9) early, strong enhancement, and (10) malignant lymphoma and inflammatory masses for differential diagnosis. A combination of these individual points yields higher sensitivity and specificity for tissue specific diagnosis and differentiation between benign and malignant lesions. Although pathology will always remain the gold standard in the diagnosis of soft tissue tumors, prediction of a specific histologic diagnosis remains one of the ultimate goals of each imaging technique. Moreover, decisions regarding biopsy and treatment could be simplified if a specific diagnosis or a limited differential diagnosis could be provided on the basis of imaging. PMID- 11431934 TI - Diagnosis of nutcracker syndrome of the left renal vein: value of the corticomedullary phase of helical CT. PMID- 11431935 TI - [Angiographic findings of hemodialysis access insufficiency]. AB - One hundred forty-six angiographic findings of malfunctioning internal arteriovenous fistulas without any history of interventional procedures were reviewed. Angiographies demonstrated 110 cases of stenosis, 72 of occlusion, 5 of venous varicosity, and 13 of venous aneurysm. Of 182 stenotic lesions, 118 (65%) lesions (74 venous stenoses: 67%; 44 venous occlusions: 61%) were revealed within five centimeters of anastomoses. In 4 of 5 cases of varicosity, proximal venous occlusions were demonstrated. On the other hand, interventional procedures were performed in 81 cases of stenoses and 14 of occlusions in the manner of PTA and fibrinolysis. The initial success rate was 90% in stenotic lesions and 71% in occlusive lesions, not a statistically significant difference. Furthermore, there was no statistically significant difference in initial success rates according to anatomical location and lesion length. Although there was no statistically significant difference according to the angiographic findings of lesions, we would like to emphasize the importance of correct understanding of anatomical changes in the dialysis shunt and of early intervention to improve the initial success rate of PTA. PMID- 11431936 TI - [A study of optimum radiographic conditions on chest examinations with computed radiology (CR) in conjunction with the offset balance of patient dose and image quality]. AB - In a report of a nationwide survey on radiographic conditions of chest radiography in Japan, it was pointed out that the average entrance surface dose (ESD) of the computed radiography (CR) system was higher than that of the film screen system. It seemed important that an objective index and criteria be established for dose reduction without a loss of image quality that would interfere with diagnostic observation. For this purpose, we investigated the properties of root of mean square (RMS) granularity, since it is a dominant factor in CR image quality and strongly depends on dose. The results indicated that RMS granularity showed little dependence on tube voltage when relative exposure was kept constant and that it decreased with the increment of exposure and approached a finite minimum value in a very high exposure region. For the most frequently used radiographic conditions in Japan (120 kV, 2.5 mmAl, 200 cm SID, 10: 1 grid), the decrement in RMS granularity from 6 to 16 mAs was 0.0276 to 0.0253 (9.1%). This finding suggested that exposure exceeding 6 mAs did not improve image quality, i.e., exposure reduction down to 6 mAs would not cause a significant loss of image quality. It was therefore concluded that RMS granularity was a useful objective index by which to determine the upper limit of exposure. Use of the most frequent conditions with 6 mAs seemed to be recommendable as an initial condition for the technical optimization of CR chest radiography, since ESD under this condition was 0.265 mGy, which was approximately equal to the value of the ESD distribution of a total chest radiogram in Japan. PMID- 11431937 TI - [Three-dimensional reconstructions with volume-rendering method of the arytenoid and cricoid cartilages by means of multidetector row CT]. AB - We created volume-rendering (VR) three-dimensional (3-D) images of the larynx using a multidetector row CT. The study included 17 examinations of 15 patients. After obtaining volume data of the larynx, VR 3-D images of airways and arytenoid and cricoid cartilages were reconstructed. The threshold of airways and calcification were set as -1025/-625 HU and 80/1560 HU, respectively. The base of the vocal processes of arytenoid cartilages and cricoarytenoid joints were demonstrated at rates of 70.6% and 53.4%, respectively. VR 3-D laryngeal images were more useful in assessing anatomy than the surface-rendering images previously reported. PMID- 11431938 TI - [Review of papers on the efficacy of the treatment in sudden deafness reported between 1989 and 1998 in Japan]. AB - We reviewed hearing recovery in 3,430 patients with sudden deafness reported between 1989 and 1998 in Japan, evaluated using standards of the Research Group on Sudden Deafness of the Ministry of Health and Welfare of Japan. Complete recovery was seen in 30.8%, marked in 24.7%, slight in 23.3% and no change in 21.8%. No remarked improvement in recovery of hearing was seen in patients with sudden deafness in these 10 years. The number of patients studied in evaluating the efficacy of therapies may thus affect results. Complete recovery from sudden deafness was nearly 30% in proportion to the increase of number of patients studied in the literature. At least 200 patients should be studied to make a reliable evaluation. PMID- 11431939 TI - [Mechanism for producing a neoglottal fricative [h] in tracheoesophageal speech]. AB - BACKGROUND: Over the past 24 years, we have been performing the tracheoesophageal (TE) fistulization for voice restoration following a total laryngectomy. The principle of this technique is to divert the exhaled air through the TE fistula into the hypopharynx, where the thyropharyngeus muscle forms the retropharyngeal prominence on which the neoglottis is located. In TE speech, the electromyographic (EMG) activity of the thyropharyngeus muscle decreases for voiceless positive production. This neoglottic articulatory adjustment plays an important role in opening the neoglottis. It has been previously reported that the glottis is open for the glottal fricative ([h]) sound produced with laryngeal articulatory adjustment and that turbulence through the glottis produces the [h] sound in laryngeal speech. Of all the voiceless consonants in the Japanese language, [h] is the most difficult to produce in TE speech. This suggests that other adjustments in addition to the neoglottic articulatory adjustment are necessary to produce the [h] sound in TE speech. This study was designed to clarify the mechanism by which the neoglottal fricative ([h]) sound is produced. SUBJECTS AND METHODS: Eight speakers who could pronounce [h] were included in this investigation, a fiber optic examination of the neoglottis, aerodynamic study, and an EMG examination of the thyropharyngeus muscle during the production of words containing voiceless fricative [h] and [s] sounds were performed in each subject. The two groups were classified according to the subject's success or failure in producing words with initial [h] and medial [h] sounds. Out of 80 TE speakers who could not pronounce [h], 4 subjects were selected as the control group. RESULTS: Fibroptic examination revealed a transient neogottal opening during the production of [h] and [s] sounds. The median of the average opening time was 0.26 seconds for initial [h] sounds and 0.19 seconds for medial [h] sounds. No significant difference in opening time was observed for [h] and [s] sounds. The aerodynamic investigation demonstrated a transient increase in supraneoglottal pressure and airflow through the neoglottis and a transient decrease in subneoglottal pressure for [h] production, while no changes were observed in the control group. The EMG study demonstrated that the activity of the thyropharyngeus muscle decreases in cooperation with a transient decrease in subneoglottal pressure only for [h] production. As a result of the transient decrease in subneoglottal pressure, the EMG activity for [h] decreased more strongly than that for [s]. CONCLUSIONS: These findings indicate that both neoglottic adjustment and pulmonary adjustment are required for [h] production in TE speech. PMID- 11431940 TI - [Differentiation and proliferation of mast cells in nasal mucosa]. AB - We determined the distribution of mast cells in nasal mucosa and studied their proliferation. Inferior turbinate mucosa was sampled in 13 patients with allergic rhinitis (allergic group) and 5 without (non allergic group) and stained immunohistochemically using anti mast cell tryptase antibody, anti-c-kit antibody, anti-PCNA antibody, and anti mast cell chymase antibody. Tissue sections stained with anti tryptase antibody revealed a higher degree of infiltration of tryptase-positive cells, i.e., mast cells, in the allergic group than in non allergic group. In the allergic group, the number of tryptase positive cells, c-kit-positive cells, and PCNA-positive cells was significantly greater in the epithelium and shallow lamina propria than that in the deep lamina propria. Tryptase-positive, c-kit-positive cells, i.e., c-kit-positive mast cells, and tryptase-positive, PCNA-positive cells, i.e., PCNA-positive mast cells, were also abundunt in the epithelium and shallow lamina propria. The stem cell factor and c-kit receptor are reported to play a primary role in mast cell differentiation and proliferation. PCNA-positive cells represent actively proliferating cells. Based on the above, we concluded that mast cells in the epithelium and shallow lamina propria in the allergic group differentiated and proliferated more actively than those in the deep lamina propria. PMID- 11431941 TI - [Speech perception ability in a patient with a 8-channel auditory brainstem implant]. AB - Auditory brainstem implant (ABI) is a central prosthesis that directly stimulates the cochlear nucleus in the brainstem for those who have interrupted auditory nerves and cannot benefit from the cochlear implantation. Speech perception in a recipient of the Nuclues 8 channel ABI, the first in Japan, is reported. A 25 year-old man with bilateral acoustic nerve tumors postlingually deafened due to tumor resection received auditory sensations with 5 channels. The correct answer using a coding strategy, SPEAK, was 35% for 5 vowels and 36% for 5 monosyllables. The use of ABI also improved his lip-reading ability on monosyllables and open set words. This indicated that he benefited from ABI, although it was limited. Even after 1 year and 3 months of follow-up, he had no serious side effects such an infection or implant rejection. PMID- 11431942 TI - [Bilateral primary carcinoma of the external auditory meatus]. AB - We report an extremely rare case of bilateral primary carcinoma of the external auditory meatus. A 50-year-old man suffered 1 month from left-ear discharge and otalgia. Examination revealed a tumor of the left ear canal and a biopsy showed well-differentiated squamous cell carcinoma. The patient underwent 60 Gy radiotherapy and left subtotal temporal bone resection. A tumor with irregular swelling of the right external canal was found 8 months after the first diagnosis. Biopsy of the right external canal confirmed the same squamous cell carcinoma. Genetic examination that the carcinoma of the right ear was probably not a metastatic from the carcinoma of the left ear. PMID- 11431943 TI - Transmediastinal approach to exploring the lung contralateral to the thoracotomy site. AB - OBJECTIVE: The approach to contralateral lung through the mediastinum is assumed useful in managing oddly distributed bilateral lung tumors. SUBJECTS AND METHODS: To remove a tumor located in the contralateral lung, a transmediastinal approach from the thoracotomy site to the contralateral lung was used in 6 patients having oddly distributed bilateral lung tumors, 1 of which was located in the contralateral lung close to the anterior or posterior mediastinum. RESULTS: All cases were treated successfully. One patient required an additional small incision on the contralateral anterior chest wall to insert an endoscopic stapler without intraoperative postural change. The postoperative course was uneventful and, to date, no local recurrence has been seen at the resected margin of the contralateral lung. CONCLUSION: This novel approach is useful, offering the advantages of reduced invasiveness and pain, shorter surgical duration, and favorable cosmetic results for patients with a tumor close to the mediastinum in the contralateral lung. PMID- 11431944 TI - Effect of tranexamic acid on blood loss reduction after cardiopulmonary bypass. AB - OBJECTIVE: We evaluated the effect of tranexamic acid on blood loss in patients undergoing elective cardiopulmonary bypass for coronary artery bypass surgery. METHODS: We randomly assigned 7 of 14 patients to a group receiving 50 mg/kg tranexamic acid before skin incision and after the start of cardiopulmonary bypass and the other 7 as controls. RESULTS: Intraoperative and postoperative blood loss was significantly (p = 0.025) reduced in the tranexamic acid group. A similar decrease in platelet count was observed during cardiopulmonary bypass in both groups. Antithrombin III was significantly (p = 0.013) decreased in both groups during cardiopulmonary bypass. Antithrombin III and thrombin-antithrombin III complexes were significantly (p = 0.001) increased after protamine administration. A significant (p = 0.010) decrease in alpha 2-plasmin inhibitor was noted at 5 and 60 minutes after the start of cardiopulmonary bypass in the tranexamic acid group. alpha 2-plasmin inhibitor-plasmin complexes were significantly (p = 0.001) increased after the start of cardiopulmonary bypass in both groups and were significantly (p = 0.012) decreased after protamine administration. alpha 2-plasmin inhibitor-plasmin complexes in the tranexamic acid group were significantly (p = 0.030) lower than in controls 60 minutes after the start of cardiopulmonary bypass, just prior to the end of cardiopulmonary bypass, and after protamine administration. CONCLUSIONS: These findings showed that tranexamic acid administration effectively prevented perioperative blood loss without thromboembolic complications and that tranexamic acid during cardiopulmonary bypass coordinates the anticoagulative effect of heparin and the antifibrinolytic effect of tranexamic acid. PMID- 11431945 TI - Prevention of surgical site infection by antibiotic spraying in the operative field during cardiac surgery. AB - OBJECTIVE: Despite the many procedures introduced to prevent surgical site infection during cardiothoracic surgery, serious infections still occur. We attempted to reduce surgical site infection by spraying antibiotic solution in the operative field--a procedure since introduced at 4 other Japanese institutions. METHODS: In the latter half of 1990, we began spraying an antibiotic solution of cefazolin (1g) and gentamicin (40 mg)/40 ml of saline placed in a 50 ml syringe and dispensed through an 18 G needle bent at 60 to 80 degrees to clean the wound during surgery. RESULT: No deep surgical site infections or deaths due to infection have occurred among the 502 patients undergoing cardiothoracic surgery under cardiopulmonary bypass at our hospital. This method was used in over 2,100 cases of similar procedures at 4 other institutions. There were 3 deaths due to severe surgical site infection (0.11%). At one institution treating over 1,000 cases a year, the incidence of death due to surgical site infection decreased significantly after this method was introduced. CONCLUSION: These preliminary experiences show that spraying antibiotic solution in the operative field reduces the risk of surgical site infection in cardiothoracic surgery. PMID- 11431946 TI - Impact of immediate cerebral angiography for in-hospital cerebral thromboembolism after cardiovascular surgery. AB - OBJECTIVES: Postoperative stroke remains a serious problem in cardiovascular surgery. We studied the role of cerebral angiography in postcardiotomy stroke. METHODS: We retrospectively analyzed 5 in-hospital patients with stroke due to cerebral thromboembolism after cardiovascular surgery. RESULTS: The incidence of in-hospital cerebral thromboembolism was 0.5%, involving 5 patients among 913 adults undergoing cardiovascular operations. In-hospital cerebral thromboembolism occurred 3 to 9 days (average: 7 +/- 2 days) after surgery. Causes of cerebral thromboembolism were chronic atrial fibrillation in 1, transient atrial fibrillation in 2, artificial valve in 1, and intracranial arterial stenosis in 1. Immediate cerebral angiography, after exclusion of intracranial hemorrhage and complete cerebral infarction by computed tomography, revealed cerebral embolism in 3 and cerebral thrombosis in 2 with occlusion or stenosis of intracranial arteries. Local intraarterial administration of a thrombolytic agent was done in the 3 patients with cerebral embolism and occluded arteries were recanalized. Fibrinolysis was not done in 2 because of recanalized arteries or sufficient peripheral blood supply through collateral feeders. No patients exhibited rebleeding into the pericardial space or wound bleeding. All patients survived with moderate or full functional recovery. CONCLUSION: Immediate cerebral angiography with/without local thrombolysis may improve functional outcome and survival in patients with postcardiotomy cerebral thromboembolism. PMID- 11431947 TI - Reduced tolerance of global ischemia in the hypertrophied heart. Effect of coronary flow regulation during reperfusion on postischemic recovery. AB - OBJECTIVE: Reduced coronary reserve during reperfusion may cause postischemic diastolic dysfunction in pressure-overload-induced hypertrophy. We studied the effect of coronary flow regulation (simulated hyperemic or depressed flow) on postischemic cardiac function during reperfusion. METHODS: Left ventricular pressure overload was induced in 4-week-old rats by abdominal aortic constriction. At 6 weeks of age, isolated Langendorff-perfused hearts (perfusion pressures: 75 mmHg in controls and 110 mmHg in the aortic constriction group) were subjected to hypothermic global ischemia (15 degrees C, 210 min), followed by 2 types of coronary flow regulation during the initial 20 min of reperfusion- manipulated high flow in control hearts (group I), manipulated low flow in control hearts (group II), manipulated high flow in aortic constriction hearts (group III), and manipulated low flow in aortic constriction hearts (group IV) (n = 6/group), and then constant pressure perfusion during the subsequent 45 min of reperfusion. Cardiac function was measured using an isovolumic balloon in the pre and postischemic periods. RESULTS: Aortic constriction hearts exhibited greater left ventricular end-diastolic pressure than did control hearts. The increase in left ventricular end-diastolic pressure did not differ between group I (3 +/- 2 mmHg) and group II (-1 +/- 1 mmHg) or between group III (29 +/- 5 mmHg) and group IV (30 +/- 6 mmHg). No difference was seen in postischemic recovery of left ventricular systolic pressure between high and low flow groups in control and aortic constriction hearts. CONCLUSION: Manipulations in coronary flow during reperfusion did not affect postischemic cardiac function in control or aortic constriction hearts, suggesting that depressed coronary flow during early reperfusion is not a primary cause of postischemic diastolic dysfunction in the hypertrophied myocardium. PMID- 11431948 TI - A modified "elephant trunk" procedure for aortic arch replacement. AB - OBJECTIVES: We have devised a modified "elephant trunk" technique for primary aortic arch replacement that strengthens the distal anastomosis by preventing access of the blood stream to the distal suture line. The technique entails suturing an elephant trunk graft to the inner surface of the arch graft circumferentially just distal to the fourth of its 4 limbs. The inner graft is then inserted into the distal aortic lumen while the edge of the arch graft orifice is anastomosed to the aortic stump. This clinical trial was conducted to clarify its usefulness. METHODS: This technique was used in 5 cases of total arch replacement to reinforce distal anastomoses. RESULTS: All patients tolerated surgery well without complications such as paraplegia or thromboembolism. CONCLUSIONS: The advantages of this modified technique include simplicity and reproducibility. PMID- 11431949 TI - Bullectomy is beneficial in the pulmonary blood flow/driving pressure relationship in bullous emphysema patients. AB - OBJECTIVES: Few physiological studies have been on pulmonary circulation, including the relationship between pulmonary blood flow and driving pressure, and viscosity changes arising from hematocrit changes before versus after lung volume reduction Surgery or bullectomy in emphysematous patients. METHODS: We studied the relationship between pulmonary blood flow and driving pressure before and after bullectomy using a unilateral pulmonary artery occlusion test based on pulmonary blood flow fractions obtained by pulmonary blood flow scintigraphy in 7 patients with pulmonary emphysema associated with bullae. RESULTS: Among the 7 undergoing bullectomy, the pulmonary blood flow/driving pressure relationship decreased in 5 and was unchanged in 2. Of the 11 lungs treated by bullectomy, this relationship decreased in 8 lungs, but not 3 (p < 0.01). No significant difference was seen in oxygen partial pressure in arterial and mixed venous blood or hematocrit. CONCLUSION: We concluded that bullectomy shifted the pulmonary blood flow/driving pressure relationship downword in some cases with significant bullae. PMID- 11431950 TI - Optimal positioning of temporary epicardial atrial pacing leads after cardiac surgery. AB - OBJECTIVES: Atrial pacing plays an important role in preventing low output syndrome and arrhythmia after cardiac surgery. We studied the optimal positioning for temporary epicardiac atrial pacing. METHODS: The performance of temporary epicardiac atrial pacing leads was examined after 13 cases of elective coronary artery bypass grafting between October 1999 and January 2000. Two bipolar electrode leads were used--1 on the cephalic atrial wall between the left and right atrial appendages, where the Bachmann bundle indwells (site A), and the other on the interatrial groove (site B). To assess pacing performance on postoperative days 1, 2, and 7, we measured 3 pacing patterns--bipolar use of sites A and B leads, and combined use of the 2 with the site A lead acting as the negative electrode and the site B lead as the ground. The pacing threshold was measured at a 0.5 ms pulse width, using the P wave amplitude and slew rate as indicators of sensing performance. RESULTS: Bipolar pacing both at sites A and A B was superior to that at site B in pacing threshold and sensing parameters. CONCLUSIONS: The negative electrode at site A is mandatory for high atrial pacing and sensing performance. PMID- 11431951 TI - Nitinol stent for the treatment of tracheobronchial stenosis. AB - OBJECTIVE: The purpose of this study was to evaluate the potential utility of implantation of a nickel-titanium alloy (nitinol) stent for the treatment of malignant or benign tracheobronchial stenosis. METHODS: We evaluated 18 patients (14 men and 4 women) who received 24 nitinol stents, between November 1997 and May 2000. All 18 patients had severe dyspnea caused by tracheobronchial stenosis. The underlying condition was malignant disease in 15 patients, and benign tracheal collapse in the other 3 patients. RESULTS: Implantation of the stent was successfully performed in all patients. Seventeen patients experienced immediate clinical improvement in respiratory symptoms. The remaining 1 patient with a bronchial fistule after lobectomy did not benefit, and died of pneumonia at 16 days after the implantation. In 15 patients, the procedure was performed using a flexible bronchoscope under local anesthesia alone, while the remaining 3 patients needed intravenous sedation. There was no complication resulting from the stent implantation. Among the 3 patients with benign tracheal collapse, 2 patients were alive at 746 and at 401 days after the stent implantation, at the time of this report. One patient with cicatricial stenosis after intubation died of heart failure due to previous myocardial infarction. Among the 15 patients with malignant disease, 4 patients have survived for 177 to 305 days to date, while the other 11 patients have died of primary malignancy with a mean survival duration of 60.2 days. CONCLUSION: The nitinol stent was effective in treating malignant or benign tracheobronchial stenosis, and had some remarkable advantages compared with other tracheobronchial stents. In stenting, most procedures can be performed using flexible bronchoscope under local anesthesia. PMID- 11431952 TI - Cardiac displacement after lobectomy in a patient with a congenital complete left sided pericardial defect. AB - A 74-year-old man underwent left upper lobectomy for a suspected lung cancer. At thoracotomy, the pericardium was found to be completely absent. The heart was fixed in a central position with an artificial patch to prevent displacement. However, the heart became displaced just after closure of the thoracotomy. The thoracotomy was immediately reopened. The anchor points of the patch on the adventitia of the descending aorta had pulled through. The heart was fixed to the surrounding mediastinal tissues with a larger patch using pledget-reinforced mattress sutures. Because simple closure may result in cardiac herniation due to the cardiac pulsations, congenital pericardial defects should be securely repaired when encountered. PMID- 11431953 TI - Oblique aortic valve replacement and coronary artery bypass grafting for severely calcified narrow aortic root with unstable angina. AB - We report an 84-year-old woman diagnosed with aortic stenosis and regurgitation with a severely calcified narrow aortic root and left main coronary artery trunk stenosis with triple-vessel coronary artery disease. Emergency aortic valve replacement and triple coronary artery bypass grafting were successful. The aortic annulus was small and heavily calcified, and the ascending aorta, the sinus of valsalva and the anterior leaflet of the mitral valve were severely calcified. A St. Jude Medical valve 19A (St. Jude Medical Inc., St. Paul, MN) was inserted obliquely along the noncoronary sinus. This technique is a useful alternative in cases where the patient's life is at risk in situations involving severe extensive calcification of a narrow aortic root. PMID- 11431954 TI - Aortic subannular left ventricular aneurysm in a patient of Asian ancestry. AB - An asymptomatic 50-year-old Japanese man was diagnosed with an aortic subannular left ventricular aneurysm during a routine physical checkup. Operative findings showed the subaortic aneurysm had developed beneath the noncoronary cusp of the aortic valve and expanded into the epicardium between the aortic root and left atrium. The operation involved patch closure of the orifice of the annular subaortic aneurysm, aortic valvuloplasty, and plication of the dilated ascending aorta. PMID- 11431955 TI - Thymic carcinoma with tumor thrombus into the superior vena cava. AB - Tumor thrombus into the vena cava have been reported in cases with renal cell carcinoma, thyroid tumor and in those with thymoma. These tumors are frequently invasive and continuous from the main tumor that shows direct vessel wall invasion. Here, we report a case of thymic carcinoma with superior vena cava syndrome, which was caused by a tumor thrombus in the superior vena cava without vessel wall invasion. The main mediastinal tumor did not show innominate vein invasion, and the superior vena cava syndrome was a result of separate tumor thrombus that was free of vessel wall invasion. The tumor thrombus could be removed through a simple venotomy. To prevent stenosis in the superior vena cava and the left innominate vein, we used a pericardial patch to close the venotomy site. PMID- 11431956 TI - Bronchial artery aneurysm treated with percutaneous transluminal coil embolization. AB - We present a case with asymptomatic mediastinal bronchial artery aneurysm at the origin of the left bronchial artery. This bronchial artery aneurysm was successfully treated with percutaneous transluminal coil embolization. Complete embolization of the feeding artery and tributaries from the aneurysm is the key to successful isolation of a large bronchial artery aneurysm from the circulation. Percutaneous transluminal embolization is one of the treatment options for a patient with asymptomatic, incidentally-identified bronchial artery aneurysm. PMID- 11431957 TI - Thrombus on the intraluminal felt strip. A possible cause of postoperative stroke. AB - We report a case of a 60-year-old woman who received an operation for acute aortic dissection and who had a postoperative complication of multiple cerebral infarction. Through aggressive investigation using transesophageal echocardiography, a mobile thrombus on the intraluminal felt strip used for the enforcement of the dissecting aortic wall was detected as the possible source of the cerebral thromboembolism. After anticoagulation therapy was started, the mobile thrombus growing on the intraluminal felt strip disappeared, and no new lesions of cerebral thromboembolism occurred. PMID- 11431958 TI - Effects of stimulus intensity on sound localization in the horizontal and upper hemispheric median plane. AB - Sound lateralization ability with headphones is known to be affected by stimulus intensity. However, there have been few studies on sound localization with loudspeakers in free field from the standpoint of stimulus intensity. The aim of this study is to obtain basic data for the tests with normal hearing and hearing impaired subjects, and for the fitting of a hearing aid. The author investigated the relationship between stimulus intensities and sound localization ability of the broadband noise and pure tone bursts delivered in the horizontal and upper hemispheric median planes. Subjects were ten volunteers, 5 males and 5 females, at the age of 18-19 years, with normal hearing. The results showed well how subjects perceived directions at all stimulus intensities. The percentage of correct response (%correct) for broadband noise stimuli was maximum at the intensity from 40 to 70 dB SPL (sound pressure level), and was minimum at 0 to 20 dB SPL in both the horizontal plane and the upper-hemispheric median plane. At lower stimulus intensity, examinee could not judge the direction, even if they could hear the stimuli. For pure tone bursts stimuli in the upper-hemispheric median plane, %correct was low irrespective of stimulus intensity. PMID- 11431959 TI - [Results of treatment of out-of-hospital cardiopulmonary arrest patients transferred to the UOEH hospital]. AB - We have accepted patients suffering out-of-hospital cardiopulmonary arrest (CPA) since September 1997. In this study, we retrospectively reviewed the etiology of CPA and the outcome of the patients. We also investigated the effects of the pre hospital cardiopulmonary resuscitation (CPR) by a bystander or emergency life saving technicians (ELST) on the restoration of spontaneous circulation. Seventy seven patients were transferred to our hospital from September 1997 to April 2000, 10 of who underwent bystander (basic) CPR. All the patients underwent advanced CPR by the attending physician immediately after admission to the emergency room. The etiology of CPA was categorized into three groups: (1) unknown endogenous disease (43 patients, 56%), (2) exacerbation of previous disease such as heart, respiratory or cerebrovascular disease (12 patients, 15%), (3) accident or suicide (22 patients, 29%). Spontaneous circulation of 30 patients was restored temporary, but only two patients recovered fully and were discharged from the hospital. The rate of restored spontaneous circulation of the CPA patients with bystander CPR was higher compared to that of the CPA patients without bystander CPR. The two fully recovered patients underwent bystander CPR. The CPR by ELST did not affect the resuscitated rate significantly. These results indicate that the most common etiology of CPA is unknown endogenous disease, and the rate of full recovery is very low (2.6%). This study suggests that bystander CPR may contribute to an improved outcome of the patients suffering out-of hospital CPA. PMID- 11431960 TI - [Involvement of orexins/hypocretins in multiple physiological functions]. AB - Orexins (orexin-A and orexin-B)/hipocretins (hypocretin-1 and hypocretin-2) are novel neuropetides discovered in 1998. Orexin-producing neurons are exclusively distributed in the lateral hypothalamic area (LHA), the posterior hypothalamic area and the perifornical nucleus in rats. The LHA is known to be a feeding center in the hypothalamus. Central administration of orexins stimulated feeding in rats and mice. Therefore, orexins are recognized as potent orexigenic peptides. In a positional cloning study, genetically narcoleptic dogs had a deletion of the orexin receptor 2 gene, resulting in a truncated, nonfunctional receptor. Orexin knockout mice were found to exhibit narcoleptic behavior. In human narcoleptic patients, hypocretin-1 in cerebrospinal fluid was undetectable in almost all patients with narcolepsy. Although the physiological role of feeding seems to be independent of sleep, it is very interesting that orexins may be common bioactive substances that are involved in regulating feeding and sleep. The discovery of orexins may contribute to clarify the pathogenesis of disorders in feeding and sleep and to develop drugs for those disorders. PMID- 11431961 TI - [Three cases of occupationally induced herpetic whitlow in health care workers]. AB - Herpetic whitlow is one of the herpes simplex virus (HSV) infections, and is known as an occupational disease. In Japan, however, there have been only a few reports of this disease. Recently, we experienced three cases of herpetic whitlow in health care workers. They presented painful vesicles on the right ring finger, the left thumb and the left index finger, respectively. All of them were considered to have been infected by HSV when they were engaged in mouth care of inpatients. We reviewed these three cases from the aspect of occupational medicine, and propose the following three points for preventing this serious nosocominal infection to health care workers: 1) re-education and enlightenment for the workers regarding hazardous behavior of HSV, 2) keeping the hands and fingers free from skin impairments, and 3) early diagnosis and treatment for herpetic whitlow of health care professionals to prevent further nosocominal infection. PMID- 11431962 TI - [Six cases of occupational skin diseases caused by cement: considerations from the aspect of occupational dermatology]. AB - Cement, in particular Portland cement, is now widely used in the field of civil engineering and the construction industry. High alkalinity of wet cement and its tiny content of water-soluble chromate can cause occupational skin diseases. In this paper, we report four cases of contact dermatitis by cement and two cases of cement burn. The occupation of the patients included two plasterers, a truck driver, a manufacturer of cement ware and two construction workers. Skin of the hands and fingers of the cement dermatitis cases was dry and fissured, and had hyperkeratotic papules and erythemas or acute exudative eczematous lesions. In one of these cases, the eczematous lesions spread to the face, extremities and trunk. All cases resulted in a positive patch testing for sodium dichromate. The cement burn cases developed severe necrotic ulcers on the leg and/or foot following prolonged contact with wet cement inside their boots. Patch testing was negative for chromate. A field trip to a construction worksite showed that method of working as well as worker's clothing at present could not thoroughly protect the skin. Therefore we concluded that better protective clothing and gloves should be used and that working conditions be improved. Most skin diseases caused by cement occur among workers at small-scale enterprises. Therefore it is desirable that regional occupational health centers, which were established to promote the health care system for workers at small-scale enterprises, take prompt measures to avoid the skin diseases. PMID- 11431963 TI - [How do universities confront and manage environmental problems?]. AB - Occupational medicine is concerned with the recognition and prevention of diseases related to the work environment. The special tools, namely, epidemiology, toxicology, and public health and clinical expertise, are joined by another specialty, environmental medicine. The Environmental Science Center (ESC) of the University of Tokyo was established in April 1975 for the purpose of treating chemically-hazardous wastes deriving from the university. The ESC houses various sections including research, education, operations, management, and also a waste-collection and-treatment division. In this review, the author intends to summarize the activity of the ESC, and then review the approaches we have applied in order to deal with the environmental problems we have faced. PMID- 11431964 TI - [Study of environmental factors for care of demented elderly men]. AB - Men interact with the environment, which sometimes leads them into distressful conditions. This stressful stimulation can induce neurodegeneration and hippocampal atrophy by increasing the glucocorticoid level. Especially, the elderly population is very vulnerable to exposure to the increased glucocorticoid due to environmental stress that results in accelerating the aging process. On the other hand, enriched physical and human environments may help to maintain their cognitive function in spite of aging. For example, aged healthy volunteers with a lower glucocorticoid level caused by avoiding distressful conditions were able to maintain large volumes of hippocampus and a better short-term memory. The purpose of this study was to investigate whether better residency for the elderly men who have a mild type of senile dementia can maintain their mental abilities. We observed and videotaped behaviors of demented elderly men who lived in either a group home or a traditional type of nursing home while they were participating in their daily activities. We made our own inventories which measured the total mental abilities, such as degrees of social interaction, degrees of motivation for participation in daily activities, degrees of independence, and degrees of facial expression, then analyzed the data with these inventories in order to compare the behaviors of the two groups. The result did not show any significant differences in the levels of mental abilities, the degrees of motivation for participation in daily activities and independence between the two groups. However, regarding social interaction, the elderly men who live in the group home exhibited a more approaching behavior toward an unfamiliar person than those who lived in the traditional nursing home. PMID- 11431965 TI - [The present situation of occupational health nursing education and how to apply it to the fundamental nursing system]. AB - The purpose of this study was to determine the present situation of occupational health nursing (OHN) education and to discuss future problems facing the fundamental nursing teaching staff. We conducted a questionnaire among fundamental nursing teachers, and our results showed that in occupational health nursing there is a gap between what is being taught at present and what the teaching staff thinks should be taught. In addition, the questionnaire showed that nursing teachers desire to teach such subjects as understanding of clients, health problems, methods of health management, the definition of OHN and concrete care in OHN, from 2 to 10 hours in the community health nursing course of the future. Because we believe that occupational health nurses can acquire a deeper knowledge and practice in continuous training, including postgraduate training, than in a fundamental nursing course, we suggest that there is the problem of how to combine fundamental nursing and continuous education in the future should be carefully studied. PMID- 11431966 TI - [Report from the 12th UOEH meeting of gastrointestinal image diagnosis]. PMID- 11431967 TI - Prompt-payment laws don't get to heart of matter. PMID- 11431968 TI - What lies beneath. Addressing hidden costs of physician/HMO conflict. PMID- 11431969 TI - National guideline clearinghouse: extensive resource underused. PMID- 11431970 TI - Speaking the unspeakable: how plans can deal with the dying. PMID- 11431971 TI - Inter-rater reliability testing for utilization management staff. PMID- 11431972 TI - 'Hospital of future' rises in Birmingham. PMID- 11431973 TI - Putting employees online worth risk of 'distractions'. PMID- 11431974 TI - JCAHO issues warning on kernicterus danger. AB - The Joint Commission is warning health care providers that a devastating yet almost always preventable condition can threaten otherwise healthy new-borns and could be considered a sentinel event. PMID- 11431975 TI - Hone your project management skills. PMID- 11431976 TI - Major review of JCAHO requirements could bring welcome changes. AB - The Joint Commission's task force for reviewing its standards is expected to produce a bounty of recommendations for improvements, but not everyone is giddy about the possibility. Though the task force is just now getting started with its work, some already are shrugging their shoulders and dismissing the effort as a lot of bureaucratic lip service. PMID- 11431977 TI - Evidence-based measures designed for diabetes care. AB - For the first time, organizations representing the perspectives of physicians, health plans, hospitals, and other health care organizations have cooperated in the development of a common set of evidence-based measures for evaluating performance in health care. PMID- 11431978 TI - Study's focus on measuring compliance, effectiveness. AB - A new program developed by Pricewaterhouse Coopers and the University of California-Los Angeles School of Public Health's Department of Health Sciences may make it possible for the first time to measure effectiveness of compliance programs in the nation's hospitals. PMID- 11431979 TI - Global DOTS Expansion Plan. Progress in tuberculosis control in high-burden countries, 2001. PMID- 11431980 TI - Tibetan population data on the multiplex short tandem repeat loci--D16S539, D7S820, and D13S317. AB - The multiplex amplification system of three tetrameric short tandem repeats loci (D16S539, D7S820, and D13S317) have been analysed extensively in various populations for forensic application. Population genetic studies were carried out for these three loci in a population sample of 129 unrelated Tibetan individuals by using a multiplex polymerase chain reaction (PCR) followed by 4% polyacrylamide gel electrophoresis (PAGE) and silver staining. All loci were in accordance with the Hardy-Weinberg expectations. The observed heterozygosities of three loci-D16S539, D7S820, and D13S317 were 73.3%, 81.4% and 80.6%, respectively. The polymorphism information contents (PIC) were 0.84, 0.80 and 0.83, respectively. Comparing with Han population, there was no statistically significant difference except for D16S539 locus. PMID- 11431981 TI - [Construction of CgA gene antisense transgenic mice]. AB - In order to get CgA gene antisense DNA transgenic mouse, we constructed the CgA gene antisense DNA plasmid pCAS2C and microinjected it into the female pronucleus of fertilized mouse eggs, and transplanted them into oviduct of the foster. Every offspring of the fosters was determined by the PCR method. The positive mice had a 300 bp DNA electrophoresis band. We selected two male positive mice from 50 offspring survived of the pseudomother. Then, two positive mice crossed with normal mice respectively to reproduce offspring of F1. All offspring of F1 were determined by PCR to select positive offspring. Positive offspring of F1 carried only one allele of pCAS2C (heterozygous pCAS2C/-). Positive F1 offspring were selfcrossed, 1/4 offspring of F2 carrying two unites of one allele pCAS2C (pCAS2C/pCAS2C) are homozygous. Then, all offspring of homozygous F2 crossed with normal mice, could produce 300 bp DNA electrophoresis band by PCR. Total RNA of brain tissue of transgenic mouse was used to RT-PCR method, the 300 bp DNA product was obtained. The result indicates that the reading frame of CgA antisense DNA of pCAS2C has expressed in the transgenic mice. PMID- 11431982 TI - [Expression of inhibin alpha and inhibin/activin beta A subunits in the developing follicles of the duck]. AB - The very sensitive quantitative competitive reverse transcription-polymerase chain reaction (RT-PCR) was used to investigate the expression of mRNA for the inhibin alpha and inhibin/activin beta A subunit in the developing ovarian follicles of the duck. The results indicated all follicles showed the expression of mRNA for the inhibin alpha and inhibin/activin beta A. The inhibin alpha subunit mRNA is expressed more abundantly than the beta A subunit in the large preovulatory follicles. Competitive RT-PCR revealed that the expression of inhibin alpha subunit is the highest in small yellow follicles (SYF), the mean relative content for the F1, F2, F3, F4/5 and LWF (large white follicle) was 0.26 +/- 0.05, 0.28 +/- 0.07, 0.57 +/- 0.12, 0.98 +/- 0.09 and 0.026 +/- 0.006, respectively, compared to a mean relative content of 1.00 for the SYF. The highest level of inhibin/activin beta A mRNA was found in the F1 follicle, the mean relative content for the F2, F3, F4/5, SYF and LWF was 0.218 +/- 0.09, 0.111 +/- 0.03, 0.058 +/- 0.011, 0.053 +/- 0.013 and 0.005 +/- 0.002, respectively, compared to a mean relative content of 1.00 for the F1 follicle. Our data suggest that the expression of the alpha subunit is reduced with follicular development whereas beta A subunit expression is dramatically enhanced, which indicates the expression of inhibin alpha and inhibin/activin beta A subunit is differentially regulated during follicular development. In addition, the highest level of beta A mRNA in F1 follicle indicates the production of dimeric inhibin and/or activin primarily occurred in the largest F1 follicle. PMID- 11431983 TI - [Preliminary studies of microsatellite markers OarAE101 and BM1329 in five sheep breeds]. AB - Two microsatellite markers OarAE101 and BM1329 which were closely linked to the fecundity gene FecB in Booroola sheep were analysed for polymorphisms in Small Tail Han sheep, Hu sheep, Charollais sheep, Ujumqin sheep, Dorset sheep, F1 (Dorset x Small Tail Han sheep). The number of alleles for OarAE101 was 5 or 4 in five sheep breeds. The number of alleles for BM1329 was all 4 in five sheep breeds. The polymorphic information contents for OarAE101/BM1329 in Small Tail Han sheep, Hu sheep, Charollais sheep, Ujumquin sheep, Dorset sheep, F1(Dorset x Small Tail Han sheep) were 0.57/0.54, 0.62/0.67, 0.61/0.59, 0.62/0.66, 0.56/0.67, 0.62/0.68, respectively. Least squares mean of litter size for OarAE101 107 bp/113 bp was significantly higher than those for OarAE101 109 bp/109 bp and 107 bp/111 bp (P < 0.05) in Small Tail Han sheep. Least squares means for litter size were not significantly different among other OarAE101 genotypes. The allele 107 bp of OarAE101 had a significant positive correlation with litter size in Small Tail Han sheep, both alleles 109 bp and 111 bp of OarAE101 had a significant negative correlation with litter size in Small Tail Han sheep. Least squares mean of litter size for BM1329 146 bp/158 bp was significantly higher than those for other BM1329 genotypes (P < 0.05) in Small Tail Han sheep. Least squares means for litter size were not obviously different among other BM1329 genotypes. The allele 146 bp of BM1329 had a significant positive correlation with litter size in Small Tail Han sheep, the allele 148 bp of BM1329 had a significant negative correlation with litter size in Small Tail Han sheep. PMID- 11431984 TI - [Genetic analysis of 35 microsatellite loci in 5 lineages of xishuangbanna miniature pig inbred line]. AB - The polymorphism of 35 microsatellites in 5 lineages of Xishuangbanna small-ear miniature pig inbred line (XMI) was analysed. Nmuber of alleles of each lineage was counted, and rates of homozygote for 35 microsatellite loci in 5 lineages were calculated. According to gene frequencies of 35 microsatellites polymorphism information content (PIC) and mean heterozygosity were calculated for each lineage, and genetic distances between these lineage were estimated. The dendrograms were obtained based on genetic distances. The results suggest that rates of homozygote in these lineage are all high, and that is the highest in lineage 151. The results also suggest that polymorphism information content and mean heterozygosity in all the lineages are low. Composition of alleles of each lineage was quite different and the genetic relationship between lineages accorded with the process of inbred line. So it is suggested that the inbreeding degree of 5 lineages of XMI are all high, and the richness of genetic diversity is lower than general commercial pig breeds. It also shows each lineage has been different groups with individual genes. PMID- 11431985 TI - [Studies of localization of black streak body gene in Drosophila melanogaster]. AB - Mutation type of body color shows frequently yellow (y), black (b) and ebony (e) in Drosophila melanogaster and locates on X, second and third chromosom respectively. A mutant of black streak (bsr) is a spontaneous mutation strain building in the laboratory in 1991. Whether this mutation strain is rediscovered on the primary style or not. We have discussed this problem with general cross breeding and complementation test. The result of complementation test indicated that it is a mutant in first filial generation with trans heterozygote and that it is few wild in second filial generation with cis teterozygote due to crossing over in crossing bsr and e. But, we believe that bsr and e is located same loci (93D2) but not the same sites. This report discussed for correlation inter-gene body color and ruleless of heredity passing down for black streak body. PMID- 11431986 TI - [The cloning and verification of the Sox9 gene of the rice field eel]. AB - The Sox 9 gene plays important roles in sex determination, which is conserved in a variety of species including mammals and chicken. The positive lambda clone of the Sox 9 gene of the rice field eel was analyzed by restriction enzymes and the restriction map was constructed. This clone was further verified to be the Sox9 gene by PCR amplification and sequencing of the HMG box region. The cloning of the Sox9 gene of the rice field eel further suggests the evolutionary conservation of this gene. PMID- 11431987 TI - [RFLP analysis of the effect of wide compatibility genes in Aus variety 'Dular']. AB - RFLP markers were employed to test whether the individuals possess wide compatibility genes (WCGs) in the progenies of Dular, which is one of the typical wide compatibility varieties in ecotype Aus rice. Meanwhile the characteristics of WCGs were also analysed. The results are shown as follows: (1) the effect of WCG at S-5 locus is better than the others, (2) the combination of different wide compatibility loci is obviously effective in increasing the spikelet fertility, (3) intraallelic interaction can cause japonica macrospore abortive, while interallelic interaction has some effects in fertility expression. PMID- 11431988 TI - [The creation of 1H chromosome-specific CAPs and ASA markers of barley]. AB - Sequence-Tagged Site(STS) marker is a kind of important marker for identifying heterogeneous chromosome on account of its simplicity, speediness and specificity. There are many STS markers in use, which changed from the RFLP and RAPD markers. But some of those from RFLP markers cannot directly be used for identifying the specific chromosomes, such as MWG913. Amplified wheat genome by MWG913, a STS marker of barley's 1H chromosome, then cloned the products. These clones were classified by restricted enzyme Taq I and then sequenced. On the basis of the results of these sequences compared with barley's (obtained from the GenBank)., two restricted enzyme specifying on 1H chromosome, EcoR I and Pvu I, were chosen to digest the PCR products of T. aestivum, Secale cereale, Hordeum vulgare, Agropyron intermedium, Haynaldia villosa, Thinopyrum elongatum and identified their specilization to Hordeum vulgare genome. Furthermore, according to the difference of Pvu I recognition site, the second PCR primers were designed and identified its specificity to Hordeum vulgare genome by PCR. Thus 1H chromosomespecific CAPs and ASA markers of barley were obtained. PMID- 11431989 TI - [Production and cytogenetics of intergeneric hybrids between Ogura CMS Brassica campestris var. purpuraria and Raphanus sativus]. AB - Crosses between Ogura CMS Brassica campestris var. purpuraria (AA, 2n = 20) and Raphanus sativus (RR, 2n = 18) were made and many intergeneric hybrids were produced. The F1 seedlings did not show chlorosis at low temperature. When red Raphanus sativus varieties were used as male parent, the leaf petiole and leaf vein of F1 plants were purple, and when white Raphanus sativus varieties were used as male parent, the leaf petiole and leaf vein of F1 plants were not purple. All the F1 plants had white flowers and normal honey glands. Male gametes of the F1 were highly sterile and female gametes of the F1 were partly fertile. Cytological studies indicated that chromosome number of the F1 was 2n = 19 as expected, the mean chromosome pairing pattern was 15.53 I + 1.34 II + 0.25 III + 0.01 IV. Most chromosomes exsistet as univalents, but there also exsisted some bivalents, trivalents and even tetravalents, suggesting that chromosome set A was partly homologous with chromosome set R. PMID- 11431990 TI - [Bt gene flow of transgeic cotton]. AB - This study was carried out to determine the gene flow of transgenic cotton under Chinese ecological environment. Transgenic cotton GK-12 containing the marker gene NPTII and Bt gene was planted in the 6 x 6 m2 plot, non-transgenic cotton CCRC 12 and Xinmian 13 were planted respectively around them. At varying distances from transgenic cotton, seeds produced by the non-transgenic cotton were collected and screened for marker gene and Bt gene using kanamycine sulphate and Dot-ELISA method. PCR technique was also used in some seeds to screen Bt gene. The result indicated that gene flow was found to be high at 0-6 m, and to decrease with distances; however gene flow occurred up to distance of 36 m from the transgenic cotton plot. Bt gene flow at 3-6 m increased with increasing the diversity of transgenic cotton in the plot, but gene flow increased little at long distance. The gene flow between species was lower than between cultivars at 0-6 m, and occurred at the distance of 72 m from transgenic plot. 72 m buffer zones would serve to limit gene flow of transgenic cotton from small-scale field test. The possibility of escapes of engineered gene to wild relatives of cotton species was also discussed. PMID- 11431991 TI - [Inferring genotype of DNA molecular marker by Bayesian theorem]. AB - Bayesian theorem is applied to infer DNA molecular marker genotype (DNA chain type) from its phenotype (electrophoresis band type). The results indicate that large difference often presents in the genotype probability of a molecular marker with incomplete genetic information when it is obtained from the assumption of independence among markers as compared with that inferred from the genotypes of the flanking markers with the complete genetic information and the recombination fractions among them based on the Bayesian theorem. Therefore, before utilizing the marker information, such as in mapping quantitative trait loci (QTL), marker assisted selection (MAS) etc., Bayes' probability of the genotype for all markers with incomplete genetic information must be calculated over the whole genome for every individual. This study provided detailed procedure for the calculation of the Bayes' probability of the unknown DNA genotype. Several extensions were also discussed for the application of the Bayesian theorem in genetics. PMID- 11431992 TI - [Cloning of gene related to salt tolerance from Sinorhizobium fredii RT19 and its expression in Escherichia coli]. AB - A 4.4 kb DNA fragment related to salt tolerance containing three open reading frames was isolated from the gene library of S. fredii strain RT19. By subcloning and functional analysis, only ORF2 related to salt tolerance was obtained. The ORF2 was ligated to expression vectors pThioHisA, B and C, respectively, and recombinant expression vectors pGA, pGB and pGC containing 1.5 kb DNA fragment related to salt tolerance were constructed. These recombinant expression vectors were transformed into E. coli DH5 alpha. Inducing by IPTG and analyzing with SDS PAGE, it was found that the fusion protein encoded by pGC was expressed, and its molecular weight was equal to the sum of thioredoxin encoded by trxA and ORF2 putative protein molecular weight. The Western blot demonstrated that the target gene was successfully expressed in E. coli. PMID- 11431993 TI - [Cloning and fusion expression of detoxifying gene in Escherichia coli]. AB - In insects, esterases play an important role in the degradation of organophosphate (OP) and Carbamate (CB) insecticides. The ability of esterase B1 to degrading OP and CB insecticides opens broad prospect of using it in programs to the insecticide pollution. OPs can be detoxified by hydrolysis of their phosphoester bonds, pyrethroids and some OPs like malathion, by hydrolysis of their carboxylester bonds, and diverse carbamate insecticides, herbicides and fungicides by hydrolysis of amide or other similar bonds. Most of the candidate bioremediation enzymes identified to date have been hydrolases. In recent years, we have focused our work on the transfer of a detoxifying esterase B1 gene from mosquito into E. coli and explored the possibility of using the detoxified enzymes in environmental protection. 5' initial of esterase B1 cDNA was cloned by RT-PCR and sequenced consequently. After the combination of 5' initial and 3' terminal fragment of esterase B1 cDNA, the recombinant vector pET-ESTB1 was constructed and transformed into E. coli BL21. A 60 kD protein was induced by IPTG and its expression was temperature-dependent. After 12 h induction, the target protein occupied 27% of the total protein. A pure recombinant protein was obtained by purification, and was detected with 10% SDS-PAGE. The results showed that 22.1% of malathion was degraded by crude detoxification enzyme in 15 mins, demonstrated a high degradation property. This research provides a novel approach, which takes the advantages of eucaryotes for bioremediation of pesticide pollutions. PMID- 11431994 TI - FMD plan wins wide support. PMID- 11431995 TI - Surgical repair of skull fractures in four horses using cuttable bone plates. AB - Three horses with severely comminuted, open facial bone fractures and one horse with a comminuted, open orbital rim fracture were referred for treatment. Severe facial bone asymmetry and epistaxis were apparent in all cases and subcutaneous emphysema was present in two, however physical and neurological examinations were otherwise normal. Radiography and endoscopy were of some use in assessing the degree of damage, although the true extent of the damage was more apparent at surgery. Surgical reduction of the fractures was recommended to maximise cosmetic and functional outcome. After surgical debridement and reduction of the fractures the bone fragments remained unstable and were not amenable to stabilisation with interfragmentary wires alone, so 2 mm cuttable bone plates were used to maintain fracture alignment. Screw migration occurred in three horses and in one of these horses the plate had to be removed. Other complications were minor and in all horses the fractures healed with good cosmetic and functional outcome. The use of cuttable bone plates should be considered as a reasonable alternative to inter fragmentary wiring for unstable, comminuted fractures of the facial bones, even where fractures are open. PMID- 11431996 TI - Myelodysplasia, hypophosphataemia, vitamin D and iron deficiency in an alpaca. AB - A pregnant 2-year-old alpaca was presented for evaluation of progressive weight loss, decreased appetite and lethargy that developed in winter. Haematologic and serum biochemical analyses revealed marked anaemia, leukopenia, severe hypophosphataemia and mild hypocalcaemia. Evaluation of bone marrow core biopsies and aspirates revealed an increased proportion of immature haematopoietic cells, without sufficient numbers of blast cells to be termed an acute myeloid leukaemia (AML). 1 The haematological and bone marrow findings were suggestive of myelodysplastic syndrome (MDS). The anaemia, leukopenia, lethargy and weight loss remained refractory to medical therapy and the alpaca was euthanased on humane grounds. PMID- 11431997 TI - The use of ultrasound in the investigation of gastric carcinoma in a dog. AB - Gastric neoplasia accounts for less than 1% of all canine malignancies. Malignant epithelial tumours are the most common gastric neoplasm in dogs and are referred to as carcinoma or adenocarcinoma. Dogs with gastric carcinoma usually present with vomiting, anorexia, and weight loss. The duration of clinical signs is from weeks to months. 1 Survey and contrast radiography, endoscopy, and ultrasonography have been used in the diagnosis of gastric carcinoma in dogs. This report describes a case of gastric carcinoma in which the survey and contrast radiographs and endoscopic findings were normal. Gastric neoplasia was suspected on ultrasound examination and confirmed histologically. PMID- 11431998 TI - Hypercalcaemia associated with chronic lymphocytic leukaemia in a Giant Schnauzer. AB - A 7-year-old male Giant Schnauzer was referred with a history of severe vomiting, lethargy, weight loss, polydipsia and polyuria. Detailed investigations revealed leucocytosis with a marked lymphocytosis, mild non-regenerative anaemia, thrombocytopenia, hypercalcaemia and azotaemia. Circulating lymphocytes were small and well-differentiated, and the same lymphoid population was present in bone marrow. Chronic lymphocyctic leukaemia with associated paraneoplastic hypercalcaemia was diagnosed. Immunohistochemical staining of a bone marrow biopsy revealed a neoplastic B-cell line expressing CD79. The dog responded to therapy with prednisolone and chlorambucil for a period of 8 months. PMID- 11431999 TI - Clinical experience with sevoflurane in dogs. PMID- 11432000 TI - Controlling foot-and-mouth disease with vaccines? PMID- 11432001 TI - Prevalence of antibodies of Leptospira serovars in beef cattle in central Queensland. AB - OBJECTIVE: To obtain up-to-date data on the prevalence of antibodies to Leptospira serovars in central Queensland beef herds preliminary to assessing their role in bovine subfertility and the role of cattle as a zoonotic reservoir. DESIGN: Sera from 2857 female cattle in 68 central Queensland beef herds were tested for antibodies to 14 Leptospira serovars using the microscopic agglutination test. Vaccination use and age of cattle were collected to enable the calculation of crude and age-stratified seroprevalences. RESULTS: The most commonly detected antibodies were to serovars hardjo (15.8% crude seroprevalence), tarassovi (13.9%), pomona (4.0%) and szwajizak (2.2%). Vaccinates were omitted from the hardjo and pomona seroprevalence data. The seroprevalence for hardjo and pomona tended to increase with age of the animals. CONCLUSION: These results are broadly similar to those of previous serological surveys. The data suggest that serovars other than hardjo, pomona and tarassovi, are unlikely to have a significant role in bovine subfertility and that cattle are unlikely to be a source of human infection with them in central Queensland. PMID- 11432002 TI - Epidemiology and diagnosis of subclinical iodine deficiency in crossbred cattle of Punjab. AB - OBJECTIVE: To determine the prevalence of iodine deficiency in crossbred cows (taurus x indicus) of Punjab and to assess the diagnostic value of the plasma concentrations of thyroid hormones, total cholesterol and free fatty acids in iodine deficiency. The efficacy of ethiodised oil, as a long-term supplement of iodine, was also evaluated. PROCEDURES: Base-line survey of iodine status was conducted by measuring plasma inorganic iodine (PII) concentration in 48 crossbred cows from 31 dairy units. Concentrations of plasma triiodothyronine (T3), thyroxine (T4), total cholesterol and free fatty acids were compared between iodine deficient and normal cows. Circulating T4 and T3 concentrations were also determined before and after injection of ethiodised oil in cows with low PII levels. RESULTS: The mean prevalence of iodine deficiency was 35.9% and showed considerable geographical variation from 0 to 86% within Punjab. Cardinal clinical signs of iodine deficiency were absent and basal plasma T3 and T4 concentrations and their ratio did not differ between deficient and control cows. Activity of circulating T4 and T4:T3 ratio increased in response to ethiodised oil injection and persisted beyond 70 days after injection. CONCLUSION: Asymptomatic iodine deficiency is prevalent in crossbred cows of Punjab and response to iodine supplementation is the most reliable index of thyroid dysfunction associated with iodine deprivation. The results of this study also support the view that injection of 1 mL of 78% ethiodised oil can prevent iodine deficiency for more than 70 days. PMID- 11432003 TI - Risk factors for dystocia in farmed red deer (Cervus elaphus). AB - OBJECTIVE: To analyse data from a study of deer herd health and productivity to identify potential risk factors for the occurrence of dystocia. DESIGN: A 2-year longitudinal observational study of 14 red deer farms was carried out in New Zealand. About 2700 hinds were monitored each year for individual characteristics including body condition score (BCS), and reproductive performance. Grazing and calving management practices were recorded. During calving, farmers recorded calving difficulties (dystocia) that needed human assistance or caused the death of the hind. Risk factors potentially affecting the occurrence of dystocia were identified and analysed by multivariable statistical techniques. RESULTS: The percentage of hinds with dystocia was 0.52% (95% CI 0.32 to 0.78). Hinds with a post-winter BCS over 3.5 were 2.7 times as likely to experience dystocia as hinds scoring 3.5 or less. Given the same BCS class, bodyweight gain in late pregnancy was associated with an increased risk of dystocia. Grazing steep paddocks in the last trimester of pregnancy decreased the risk of dystocia in hinds with a BCS over 3.5. CONCLUSIONS: The risk of dystocia on red deer farms may be reduced if key individual animal, management and environmental factors are optimal. PMID- 11432004 TI - Surveys to assess the amount of pesticide in wool and the use of pesticides by woolgrowers in Queensland. AB - OBJECTIVE: To measure the amounts of pesticide residue in wool grown in Queensland between 1997 and 1999, and to describe the use of pesticides for the control of lice infestations and blowfly strike in Queensland sheep flocks. DESIGN: Pesticide residues were measured in a random sample of wool from Queensland clips offered for sale during 1998 and 1999. Information on pesticide use was obtained from a trace-back postal survey. PROCEDURE: Samples taken from wool lots were tested for the presence and amount of organophosphorus (OP), synthetic pyrethroid (SP) and insect growth regulator pesticides. A questionnaire seeking information on flock characteristics and pesticide use was sent to the manager of each flock from which a wool sample was tested. RESULTS: The mean amount of OP and SP residue was 2.0 and 0.8 mg/kg, respectively. Ninety-five and 98% of wool samples contained < 9 mg/kg and 7 mg/kg of OP and SP residues, respectively. The mean amount of cyromazine, diflubenzuron and triflumuron was 12.7, 5.8 and 13.0 mg/kg, respectively. The amounts of OP, cyromazine and triflumuron residues were greater in wool from flocks located in southern Queensland. CONCLUSION: Most (95%; 95% CI, 92-97%) Queensland wool clips grown between 1997 and 1999 meet the suggested Australian maximum acceptable residue amounts for OP and SP pesticides. PMID- 11432005 TI - Bovine nasal eosinophilic, granuloma with blood eosinophilia caused by Nocardia species. AB - OBJECTIVE: To describe a case of nocardial nasal granuloma in a Holstein heifer. CLINICAL FEATURES: The heifer showed laboured respiration and occasional coughing with granulomatous excrescences on the bilateral nasal mucosa, and a marked leucocytosis with eosinophilia. NECROPSY: Severe granulomatous inflammation with eosinophils, epithelioid cells, giant cells and Nocardia sp obstructed both nasal passages completely. CONCLUSION: This lesion resulted from chronic infection with Nocardia sp and is distinct from other forms of bovine nasal granuloma, also called atopic rhinitis or chronic granular rhinitis, which we propose should be classified as bovine allergic granular rhinitis. The most appropriate name for the present disease is bovine nasal eosinophilic granuloma. PMID- 11432006 TI - It's all a question of service. PMID- 11432007 TI - QA&CE: needs of female GPs in rural areas. PMID- 11432008 TI - Asthma in children. PMID- 11432009 TI - Transmission of chickenpox from Varicella zoster vaccination is possible. PMID- 11432010 TI - Is screening for depression worthwhile? PMID- 11432011 TI - The role of carers. PMID- 11432012 TI - The role of the research nurse. PMID- 11432013 TI - Going where the epidemic is. Epidemiology and control of hepatitis C among injecting drug users. AB - BACKGROUND: The hepatitis C virus (HCV) is continuing to spread among injecting drug users (IDUs) in Australia; other forms of transmission are uncommon or rare. Control of the epidemic of this potentially serious viral infection will only be achieved when transmission among IDUs is addressed. OBJECTIVE: To briefly review current knowledge about transmission and impact of HCV, so as to provide an informed basis for patient counselling. DISCUSSION: As better understanding develops about the transmission of HCV and the diseases it causes, and as treatments for chronic HCV infection and knowledge of how to target these treatments improve, our ability to tackle this epidemic, both at personal and public health levels, increases. However, the mainstay of treatment will continue to be fully informed counselling, and we urgently need expansion of current harm reduction programs to stop continued transmission among IDUs. PMID- 11432014 TI - Liver toxicity. Could this be a drug reaction? AB - BACKGROUND: Adverse reaction to prescription medications, over the counter medications and complementary medicines, are an important cause of liver toxicity. OBJECTIVE: To highlight the importance of drug induced liver toxicity in patients presenting with abnormal liver function tests. DISCUSSION: In patients who present with symptoms of liver dysfunction such as anorexia, itch or Jaundice and in whom liver function tests are abnormal, the possibility of an adverse reaction to medications should be considered. A detailed history of all medications taken in the past few months, including complementary medicines, should be obtained. After exclusion of other causes of liver disease such a viral hepatitis or gallstones, the cessation of the offending medication and the normalisation of liver tests is usually sufficient to establish a diagnosis. Occasionally, referral to a specialist and liver biopsy may be required. PMID- 11432015 TI - Viral hepatitis? Which test should I order? AB - BACKGROUND: There are a multitude of viruses that may cause hepatitis. The laboratory diagnosis of viral hepatitis is important in order to plan immediate patient management, determine treatment choices and provide patient education in order to limit transmission of infections to others. OBJECTIVE: This article outlines laboratory investigations that may be routinely ordered to assist in determining the etiology of viral hepatitis and summarises some preventive and treatment strategies that may be adopted. DISCUSSION: Investigations to determine exposure to infection are routinely performed and include simple serological tests, while tests to follow the course of infection or response to treatment may involve newer molecular techniques, including polymerase chain reaction (PCR), genotyping and viral quantification. PMID- 11432016 TI - Gallstones. Modern management. AB - BACKGROUND: Management of gallstones has changed as a result of new technologies, such as ultrasound, endoscopic retrograde cholangiopancreatography (ERCP) and laparoscopic surgery. OBJECTIVE: This paper describes the clinical situations in which gallstones occur and their natural history. This is then related to the advantages and disadvantages of ERCP, laparoscopic cholecystectomy and open cholecystectomy. DISCUSSION: Laparoscopic cholecystectomy has become the treatment of choice, but does have risk of serious complications of which the patient should be informed. Incidental gallstones should generally be left untreated. Patients presenting with biliary pain are certain to develop recurrence and require elective cholecystectomy. Those with acute cholecystitis should be managed early, with laparoscopic or open operation depending on the experience of the surgeon. Patients with obstructive jaundice can undergo laparoscopic duct exploration or have an ERCP/sphincterotomy. Those with gallstone pancreatitis should have laparoscopic cholecystectomy within the same hospital admission. PMID- 11432017 TI - Naltrexone and acamprosate. Using new drugs for alcohol dependence. AB - BACKGROUND: Two new drugs for the treatment of alcohol dependence have recently become available in Australia. Both naltrexone and acamprosate have an 'anticraving' action, but act on different pathways and have different properties. OBJECTIVE: This article aims to give an outline of the major pharmacological features of the two new drugs and some guidelines on their use for general practitioners. DISCUSSION: The past 50 years have seen major advances in almost all medical disciplines. A notable exception has been the alcohol and drug field. The arrival of methadone to Australia in the 1960s was, until lately, the most recent advance in the field. Over the past 18 months, several new drugs have become available for use in alcohol and drug (A&D) clinics. These drugs have also become available to the GP, but their use by this group remains relatively small. PMID- 11432018 TI - Regional nerve blocks. Part 1--An introduction. AB - Sustained nerve trauma is unlikely in a block. Local anaesthetics need time to work--the bigger the nerve the longer they need. Always perform a nerve block on an awake patient. Very young children and the frail elderly are more susceptible to cardiac toxicity. When used appropriately adrenaline slows the uptake of local anaesthetic into plasma. PMID- 11432019 TI - Markedly raised serum prostate specific antigen levels. Prostatic infarction rather than malignancy? AB - BACKGROUND: Though prostate specific antigen (PSA) and prostate acid phosphatase (PAP) are of greater value in the postoperative monitoring of patients with cancer of the prostate, high preoperative levels suggest the probability of malignancy. Benign conditions of the prostate are known to raise the serum levels of these enzymes, however, the higher the level of these markers the stronger is the suspicion of malignancy. OBJECTIVE: This report describes five patients who presented with acute urinary retention and had evidence of prostatic infarction and chronic prostatitis on histology. DISCUSSION: The combination of these factors probably accounted for the extremely elevated levels of PSA and PAP as other possible causes were ruled out. PMID- 11432020 TI - Migraine and hypertension. Is there a relationship? AB - BACKGROUND Migraine and hypertension are common conditions that frequently coexist. The relastionship between the two is usually coincidental, but some evidence suggests that poor control of blood pressure may exacerbate the frequency and severity of migraine. OBJECTIVE To review the relationship between migraine, other headaches and blood pressure and to discuss guidelines for suitable therapy for both conditions when they occur together. DISCUSSION Establishing the blood pressure should be a routine task in the assessment of all headache patients. Severe hypertension in the setting of new acute headache may indicate a serious underlying cause and requires urgent investigation. In patients with migraine and established hypertension, good control of blood pressure may be beneficial in controlling their headache. Many of the drugs used to treat hypertension may cause headache and some agents used to treat migraine can exacerbate hypertension and so careful consideration of the therapeutic options is important. PMID- 11432021 TI - Herbal medicine in neuropsychiatry. PMID- 11432022 TI - Herb-drug interaction guide. AB - Herbal medicines now fall into the category of complementary medicines. With their increasing popularity, a new set of circumstances has arisen--the herb-drug interaction. In an effort to streamline the interaction information available, the author has designed two charts. The first lists the 21 most popular herbal medicines sold in Australia, sourced from the largest natural supplement manufacturer in Australia, Bullivant Natural Health Products. The second lists the 12 commonly prescribed prescription drug classes. This list was sourced from F H Faulding & Co Ltd, together with Australian Prescriber. Specific interaction information was collated from a variety of sources--medical and complementary medicine journals, pharmacy practice journals and respected phytotherapy texts. These tables are excerpts from these charts. PMID- 11432023 TI - The depression puzzle. Do the pieces fit? AB - BACKGROUND: Depression is a common problem in the Australian community. It results in considerable disability. Currently screening tools promoted for detection of depression are nonspecific. They are not diagnostic tools. While people identified by screening are, by definition, at higher risk of depression, the diagnosis still needs to be based on interview findings. Screening tools do not substitute for time spent with patients developing rapport and an understanding of the causes of the depression and formulating possible management steps. Screening alone has not been shown to improve outcomes for people with depression. OBJECTIVE: This article examines the current prevalence of depression in Australia and whether screening tools ultimately contribute to a reduction in morbidity. DISCUSSION: Screening instruments have arbitrary cut off points for identifying patients with depression and their findings regarding prevalence should be applied with caution. If the thresholds are low then 'prevalences' are raised. It is unclear whether all 'cases' detected using screening instruments require clinical intervention. PMID- 11432024 TI - Food for thought. PMID- 11432025 TI - A little bit of magic. PMID- 11432026 TI - Research and evaluation. Implications for the reflective general practitioner. PMID- 11432027 TI - Randomised trials within the general practice evaluation program. Why so few? AB - BACKGROUND: The General Practice Evaluation Program (GPEP) provided competitive funding for general practice research from 1990-1999. METHODS: Projects funded by GPEP between November 1990 and December 1999 were audited to determine the extent of randomised trials and other interventional studies. RESULTS: Thirty-three (13%) of the 248 projects were interventional studies; of these 21 (8% of all projects) were randomised trials. CONCLUSIONS: While GPEP has made a significant contribution toward Australian general practice research, few randomised trials and other interventional studies have been funded. Reasons include the specific evaluative focus and restrictions on project duration and funding. Randomised trials should be supported within future general practice research funding. PMID- 11432028 TI - Establishing a practice based primary care research network. The University Family Practice Network in South Australia. AB - BACKGROUND: Practice based research networks enable the study of primary care problems in a primary care setting. To be successful, our experience indicates that a network will need to incorporate a number of key components. In South Australia, a University Family Practice Network has been established to undertake primary health care research and teaching and to contribute to the general practice workforce. The University Family Practice Network is composed of five practices and is managed jointly by the Department of General Practice, Adelaide University and the South Australian Centre for Rural and Remote Health, Adelaide University and the University of South Australia. OBJECTIVE: This article details the successful establishment of the University Family Practice Network in the hope of providing a model for other universities who may be interested in setting up a practice based research network. DISCUSSION: The characteristics of this network include fractional academic positions for general practitioners, computerised medical records, location in rural and urban sites, regular meetings and the appointment of a research fellow. A number of benefits have arisen since the network's establishment that will assist its future development and sustainability. These include collocation with allied health services, development of local expertise and workforce support. PMID- 11432029 TI - Personal doctoring. Its impact on continuity of care as measured by the comprehensiveness of care score. AB - OBJECTIVE: To determine the impact of personal provider continuity on continuity of care as measured by the comprehensiveness of care score. DESIGN: Retrospective cross sectional analysis of medical records. SETTING: The setting was a four doctor practice on the New South Wales Central Coast of Australia. METHOD: The subjects were 131 male and 123 female patients with a mean age of 42.7 years (SD 25.9) median age of 42 years and an age range of 1-95 years. The main outcome measures were a comprehensiveness score for each patient in the personal provider continuity and discontinuity of care group over a two year period. RESULTS: The overall comprehensiveness scores in the personal provider continuity group was 7.38 (95% CI: 7.04-7.71) compared to 6.03 (95% CI: 5.7-6.35) for those in the discontinuity group (p < 0.000). A linear regression model revealed that 15.8% of the total variance of the comprehensiveness score is explained by the two independent variables 'modified continuity index' (13.6%) and 'age' (2.2%). Nonrelated independent variables are gender, number of visits and number of years attending the practice. CONCLUSION: Personal doctoring significantly improves continuity of care as measured by the comprehensiveness of care score, and this observation is essentially age independent. These findings clearly suggest that patients should be encouraged to find and stay with one doctor, and that practices should develop systems to enable patients access to their usual provider. Both strategies, combined with the awareness of potential gaps in our service provision, will increase the likelihood of achieving increased continuity of care. PMID- 11432030 TI - Glatiramer acetate for multiple sclerosis. AB - Glatiramer acetate (Copaxone-Teva) was first marketed in the UK last year as a treatment for reducing the "frequency of relapses in ambulatory patients with relapsing-remitting multiple sclerosis characterised by at least one clinical relapse over the preceding two-year period". Here, we assess the place of this drug. PMID- 11432031 TI - Antimicrobial prophylaxis for orthopaedic surgery. AB - The number and range of orthopaedic procedures performed in the UK has increased over recent decades, with many more devices now being implanted to replace diseased joints, stabilise bone or correct skeletal deformities. With these advances, the number, type and complexity of potential surgical site infections have increased. Postoperative orthopaedic infections can impair the functional result of surgery, prolong hospital admission, harm patients and sometimes prove fatal. They require vigorous treatment with antibiotics and often necessitate further surgery. Here, we discuss measures for preventing surgical site infection after orthopaedic surgery, concentrating on major joint replacement and surgery for long bone fractures. PMID- 11432032 TI - Defining disease: determining practice. AB - Management of patients traditionally starts with establishing a diagnosis, a process that relies heavily on how diseases are defined. But definitions of disease vary with time and are influenced by, amongst other things, culture, politics, sociology and economics. The ninth annual Drug and Therapeutics Bulletin symposium, 'Defining disease: determining practice', explored the ways disease is defined and how this might affect practice. Here, we summarise the key points raised in the presentations and subsequent discussion. PMID- 11432033 TI - [Correlation between ABH glycolipids in sperm and functional test]. AB - OBJECTIVE: To determine the association, if any, between the human spermatic membrane integrity test and normal or altered membrane ABH glycosphingolipids expression. METHODS: Semen samples from 50 patients who consulted at the infertility services of the Provincial del Centenario and Eva Peron de Granadero Belgorria teaching hospitals in Rosario (Argentina) were collected and analyzed in accordance with WHO guidelines. The percentage of dead sperm was analyzed by the eosin test while the percentage of swollen sperm was analyzed by the hypoosmotic test. They were classified according to their ABH expression, either diminished or absent (group 1) or normal (group 2), by applying the hemagglutination inhibition and specific antibody consumption technique. For the statistical analysis the two groups were compared with both spermatic viability tests using the non parametric Mann-Whitney technique for independent samples, since the assumptions for the application of the parametric test (test t) were not fulfilled. RESULTS: A significant difference was found in the percentage of dead sperm (eosin test) of both groups (p < 0.001). However, there was no significant difference in the percentage of swollen sperm (hypoosmotic test) of the two groups. CONCLUSIONS: Our results and the data published in the literature indicate a relationship between a reduced ABH expression and spermatozoid membrane integrity, especially at the level of the head, which leads us to consider that ABH glycosphingolipids are mainly located in this segment of the human spermatozoon involved in the spermatozoon-ovum interactions. PMID- 11432034 TI - [Prostatic cysts of the midline: management and follow-up]. AB - OBJECTIVE: To analyze the presenting features, diagnosis and management of uncommon cystic lesions located at the midline of the prostate. METHODS: From 1990 to 1999, 8 patients with medial prostatic cyst were treated in our hospital. Vesicorenal ultrasound evaluation was the first procedure that was performed and was diagnostic in most of the cases. Only those patients that underwent surgery required admission to hospital. RESULTS: The most common presenting features were irritative and obstructive voiding syndromes. Treatment achieved cure or clinical improvement. CONCLUSIONS: Vesicorenal ultrasound assessment is the first procedure that should be considered in the diagnosis of this disease. Before ascribing the symptoms to a prostate cyst, other urological conditions with more prognostic relevance should be excluded. Treatment should be instituted only in the symptomatic patients and should be individualized according to the characteristics of each case. PMID- 11432035 TI - [Cytoscopic follow-up of initial G3T1 bladder tumors treated with BCG]. AB - OBJECTIVE: To evaluate the cystoscopic findings during initial follow-up, the anatomopathological correlation of tumor endoscopic features and the results of standard control multiple biopsy performed 6 months after TUR in patients with G3T1 transitional carcinoma treated with BCG. METHODS: 114 patients with G3T1 bladder tumor (52% associated with Cis) were treated with 81 mg Connaught BCG intravesical instillations weekly for 6 consecutive weeks. Follow-up was performed with cystoscopy and cytology at 3 months, and cystoscopy and standard multiple biopsy at 6 months. The endoscopic findings were described as normal bladder, macroscopically tumorous lesion or erythematous lesion. RESULTS: During the first 6 months of follow-up superficial recurrence was found in 16% and 5% showed progression to muscle invasion. Tumor recurrence or progression was found in 61% and 39% at 3 and 6 months, respectively. Most of the macroscopically tumorous lesions resulted in a tumor at 3 and 6 months in 56% and 64%, respectively, and the remaining lesions were mainly inflammatory granulomas produced by BCG therapy. Twenty biopsies of erythematous areas detected only one case of Cis (5%) and 98 standard multiple biopsies of endoscopically normal mucosa detected 10 cases of Cis (overall, 3 at 3 months and 7 at 6 months); all cases were preceded by initial Cis except in one case. CONCLUSIONS: Cystoscopy performed at 3 months is very useful since it detected 61% of the superficial recurrences and 66% of the cases with progression to muscle invasion during the first 6 months. Routine biopsy of erythematous areas detected during cystoscopy is of little value since a large number of these biopsies are unnecessary in view of its diagnostic yield (5%). Since 90% of the Cis detected during the first 6 months of follow-up were patients with Cis in the initial tumor, it would be appropriate to perform standard multiple biopsy for control only in this subgroup of patients if the sensitivity of cytology is low in high grade tumors or Cis. PMID- 11432036 TI - [Correlation between imaging findings and anatomopathologic results in radical prostatectomy. Data from our series]. AB - OBJECTIVE: To analyze the diagnostic methods utilized in prostate cancer and the preoperative information on the extent of the tumor, and compare these results with those obtained from anatomopathological analyses of the radical prostatectomy specimen. METHODS: Data on the radical prostatectomies performed during the study period were obtained from the Admissions and Clinical Records services and were analyzed using the SPSS statistical analysis software for Windows. RESULTS: From 1991 to 1998, 109 radical prostatectomies were performed. Evaluation by transrectal US was carried out in 89 patients (81.6%) and detected a tumor (unilateral or bilateral) in 77, for a sensitivity of 86% (CI 78.8-93.2). A CT study was performed in 94 patients and was positive in 25, for a sensitivity of 26.5% (CI 17.6-35.4). The sensitivity for transrectal US and CT were 4.17% and 3.33% for capsular involvement, 5.88% and 5.26% for seminal vesicle involvement, and 0% and 0% for node involvement, respectively. CONCLUSIONS: Transrectal ultrasound is a reliable diagnostic imaging method, although it has a low sensitivity when used to determine the extent of the tumor. In our series, the radiological methods showed a low sensitivity when they were utilized to determine the extent of the prostate cancer. Routine preoperative assessment by CT can be obviated in prostate cancer. PMID- 11432037 TI - [Prevalence and risk factors for erectile dysfunction in the Region of Bierzo]. AB - OBJECTIVE: To analyze the prevalence and risk factors for erectile dysfunction in Bierzo (Leon, Spain). METHODS: A study was conducted based on a self-administered survey in males aged 19 to 90 years that consulted at the urology and family medicine services of 10 health centers in Bierzo (Leon, Spain). To determine the existence or absence of erectile dysfunction, the male sexual health questionnaire (SHIM) was included, and those with a score of 21 or less were considered to have some degree of erectile dysfunction. Distribution of the questionnaire began in September 1999 and ended in May 2000. Submission of the questionnaires concluded on May 31, 2000. Of 935 questionnaire that were received, 830 were valid. RESULTS: The overall prevalence of erectile dysfunction in this study was 63.9%, which increased with age and was markedly higher after age 50 years (38.6% for age 41-50 years and 72.6% for age 51-60 years). The age adjusted analysis showed the risk factors for erectile dysfunction were depression and arterial hypertension on treatment with drugs. Diabetes mellitus on treatment with oral glucose lowering agents or insulin, heart disease and hypercholesterolemia were also associated with the more severe forms. CONCLUSIONS: The prevalence of erectile dysfunction in patients consulting at the primary care and general urological services in Bierzo (Leon, Spain) was outstandingly high. The present study showed erectile dysfunction to be an age dependent disease that caused concern in slightly more than half of the patients with this condition. Apart from age, depression and vascular disease were found to be risk factors. PMID- 11432038 TI - [Incidence of priapism at the Marques de Valdecilla University Hospital in Santander]. AB - OBJECTIVE: Our experience in the management of priapism, its etiology, blood gas changes, treatment and outcome are presented. METHODS: The records of patients that had been treated at the Urology services of our hospital over the last 8 years were reviewed. During this period, patients with erectile dysfunction were treated according to a protocol. RESULTS: There were 9 episodes of priapism in 8 patients; all cases were low flow and the patients were aged 37-71 years. The duration of the prolonged erection ranged from 4 to 72 hours. Of these 9 cases, the etiology was intracavernosal PGE in 7, after trazodone administration in one case, and unknown in one case. Intracavernosal metoxamine was administered in all cases with excellent results, except one case that required a saphenous cavernosal shunt and another case that was initially treated with adrenalin also with good results. CONCLUSIONS: Since intracavernosal agents have been utilized in the treatment of erectile dysfunction, priapism has become a more frequent urological emergency. Although a lower incidence has been reported for PGE, the dose should be well adjusted to avoid a higher incidence. PMID- 11432039 TI - [Vaginal wall sling in the treatment of female stress urinary incontinence. Analysis of 27 cases]. AB - OBJECTIVE: To evaluate the results of the vaginal wall sling procedure in the treatment of stress incontinence. METHODS: A retrospective study was conducted to evaluate the clinical outcome of the vaginal wall sling procedure in the treatment of genuine stress incontinence. The study comprised 27 women with a mean follow-up of 15 months postoperatively. RESULTS: Of the 27 patients who underwent this surgical procedure, 12 (44%) are continent and 9 (33%) have improved, accounting for 77% overall cure and improvement rate. Six patients (22%) remained incontinent. Mean operating time was 90 minutes and mean length of hospital stay was 8 days. CONCLUSION: In our experience, the vaginal wall sling procedure for genuine stress incontinence did not achieve the good results obtained in other series. PMID- 11432040 TI - [Mucinous adenocarcinoma of the prostate]. AB - OBJECTIVE: To describe a case of mucinous adenocarcinoma of the prostate. METHODS: A 77-year-old patient presented with urinary complaints. Patient evaluation showed an enlarged prostate and increased PSA levels. RESULTS: A prostate biopsy showed a glandular neoplasm with extensive mucinous differentiation and abundant lacunae of PAS positive mucus (mucinous adenocarcinoma). The patient remains well one year after the diagnosis. CONCLUSIONS: Mucinous adenocarcinoma of the prostate is a rare variant that should be recognized and does not necessarily imply a worse prognosis than other carcinomas of the prostate. PMID- 11432041 TI - [Bladder adenocarcinoma with glandular cystitis areas]. AB - OBJECTIVE: To describe a case of adenocarcinoma of the urinary bladder. METHODS: A 63-year-old patient consulted for gross hematuria. Ultrasound evaluation showed a bladder neoplasm and CT demonstrated extensive infiltration of adjacent structures. RESULTS: Histological analysis of the TUR specimen confirmed a muscle infiltrating glandular neoplasm (adenocarcinoma) with diffuse areas of cystitis glandularis. Patient evolution has been poor and he is now in the end stage of the disease. CONCLUSIONS: The morphology is not unlike that of colorectal carcinoma, which should be distinguished. The association of areas of cystitis glandularis indicates a possible relationship. PMID- 11432042 TI - [Mucinous adenocarcinoma of the prostate. Report of a new case]. AB - OBJECTIVE: To describe a case of mucinous adenocarcinoma of the prostate, an uncommon variant of prostatic carcinoma of which few cases have been reported in the literature. This tumor type is briefly reviewed with special reference to the controversial issues that remain unresolved due to the small number of cases. METHODS/RESULTS: A case of mucinous adenocarcinoma of the prostate is presented. An extraprostatic origin had been discarded. The diagnosis was made by DRE, transrectal US and histological confirmation by transrectal biopsy. PSA values were normal. Studies to determine the extent of the lesion showed advanced locoregional dissemination. Surgery was therefore not indicated and hormonal therapy was administered. Patient outcome was poor with no apparent clinical response to hormone blockade. Radiotherapy was attempted. The patient rapidly became worse and died. CONCLUSIONS: Mucinous adenocarcinoma of the prostate is characterized by the presence of significant amounts of extra and intracellular mucin. Its clinical features are not unlike those of classical adenocarcinoma and in general terms, it is considered to carry a worse prognosis and refractory to hormone therapy. PMID- 11432043 TI - [Giant orthotopic ureterocele. Diagnosis by ureteral jet detection with doppler color ultrasonography]. AB - OBJECTIVE: To present an additional case of orthotopic ureterocele in a patient that consulted for hematuria and outlet obstruction, with special reference to the utility of non-invasive color doppler ultrasound in the diagnosis of this condition. METHODS/RESULTS: This condition was suspected on detecting a ureteral jet inside a cystic lesion of the bladder. The diagnosis was confirmed by IVP and cytoscopy. Treatment was by transurethral resection with good functional results. CONCLUSION: Color doppler ultrasound is useful for the diagnosis of ureteroceles, particularly in patients in whom contrast or ionizing radiation must be avoided. PMID- 11432044 TI - [Xanthogranulomatous cystitis: report of a case]. AB - OBJECTIVE: To report a case of xanthogranulomatous cystitis, a rare disease especially in the western countries (12 of the 16 cases reported were Japanese and Korean). METHODS: A case of xanthogranulomatous cystitis in a 34-year-old female is presented. Patient evaluation included histological, histochemical and immunohistochemical studies. RESULTS/CONCLUSIONS: The lesion was localized to the bladder dome, anatomically related with the urachus. It consisted of collections of histiocytes with lymphoplasmocytic infiltrate. The adhesions observed in the omentum in this case indicate a possible relationship with a previous inflammatory process. Surgery is advocated in most of the cases with good results. PMID- 11432045 TI - [Mesoblastic nephroma in the adult: report of a new case]. AB - OBJECTIVE: A case of adult mesoblastic nephroma is presented. The clinical features, treatment, histological diagnosis and outcome are discussed. METHODS/RESULTS: A 68-year-old patient in whom a renal mass had been detected by ultrasound is described. The patient underwent radical nephrectomy. The anatomopathological analysis demonstrated a mesoblastic nephroma. No signs of recurrence have been observed at 3 years' follow-up. CONCLUSIONS: Mesoblastic nephroma of adulthood has a benign behavior and recurrence is rare after surgery. PMID- 11432046 TI - [Male pseudohermaphroditism. Report of a new case]. AB - OBJECTIVE: To report a case of male pseudohermaphroditism. METHODS: A 45 day-old infant was referred for ambiguity of external genitalia. The anatomy of the external and internal genitalia and chromosomal and gonadal sex were analyzed. RESULTS/CONCLUSIONS: Female sex was assigned and labioscrotal reduction and orchidectomy were performed at age 6 months with good adaptation to gender role. PMID- 11432047 TI - Lithogenic risk factors for renal stones in patients with Crohn's disease. AB - OBJECTIVES: Calcium oxalate kidney stones are more common in patients with Crohn's disease (CD). The aims of this study were to verify the prevalence of the main risk factors for calcium oxalate nephrolithiasis in patients with CD and to evaluate the degree of urinary relative supersaturation for calcium oxalate (CaOx), dihydrogen uric acid (DHUA) and monohydrogen calcium phosphate (MHCaP). SUBJECTS AND STUDY PROTOCOL: 42 patients with CD (22 male and 20 female, aged 15 72 years) and 18 controls (8 male and 10 female, aged 26-65) were studied. Nine patients were evaluated during an active episode and 33 in a quiescent phase. All patients had normal glomerular filtration rate. All subjects collected a 24-hr urine sample and fasting venous blood was drawn. Good compliance of urine collection was assessed by the Cockcroft and Gault formula. In urine pH and oxalate (Ox), calcium (Ca), phosphate (P), uric acid (UA), citrate (Cit), magnesium (Mg), sulphate (Sulph), sodium, potassium and chloride concentrations were measured and their excretions calculated. Urinary RS index was obtained using the software EQUIL93. RESULTS: A decreased urinary volume (61.9%) was the most frequent finding. A decreased excretion of Cit, Mg and Sulph (38.1%, 31.0% and 31.0%, respectively) and increased excretion of P, Ox, UA and Ca (33.3%, 23.8%, 16.7% and 14.3%, respectively) were found. Thirty four patients (81.0%) showed at least 2 lithogenic risk factors and only 2 patients showed none. Urine of patients had a higher urinary CaOx and DHUA relative super saturation. Patients studied in an active episode showed a higher urinary CaOx and MHCaP RS than those studied in the quiescent period. CONCLUSIONS: The majority of patients with CD have a multifactorial high risk for calcium oxalate and a single patient usually has several metabolic disturbances which are more evident in an active episode. PMID- 11432048 TI - Severe palmar hyperhidrosis treated by transthoracic endoscopic sympathectomy. PMID- 11432049 TI - The Geriatric Medicine Fellowship Program at the John A. Burns School of Medicine, University of Hawaii. PMID- 11432050 TI - Improvement in quality of life after bilateral transthoracic endoscopic sympathectomy for palmar hyperhydrosis. AB - OBJECTIVE: To evaluate the efficacy of bilateral transthoracic endoscopic sympathectomy (TES), in alleviating symptoms and improving quality of life for patients in Hawaii. DESIGN: Retrospective cohort study. MATERIALS AND METHODS: Patients who had undergone TES were evaluated by phone interview and the SF-36 questionnaire to assess improvements in symptoms and the development of compensatory hyperhydrosis. SF-36 scores were divided into 8 scales and evaluated by one-tailed t-test. RESULTS: Since 1999, eight patients (five women and three men, mean age 27.4 years old, range 15-41 yrs) underwent TES without significant complication. Length of hospital stay was less than one day for all patients except one, who stayed four days. Estimated operative blood lost was less than 100 ml and no blood transfusions were required. No Horner's syndrome was suffered. After a mean follow-up of 7.0 months (range 1.2-15.8 months), none of the patients had recurrent symptoms in the palms but all reported moderate compensatory hyperhydrosis located mainly in the trunk and lower extremities (two patients). SF-36 scores showed significant improvements in social functioning (p < 0.005), mental health (p < 0.049), and role-physical (p < 0.020) along with an increase in bodily pain (p < 0.012). CONCLUSION: Although TES resulted in some bodily pain and compensatory hyperhydrosis; these elements were outweighed by the improvement in palmar symptoms, social, mental, and role physical functioning, and overall quality of life. PMID- 11432051 TI - Skin Cancer Prevention Research Receives National Award. PMID- 11432052 TI - [Ecological industrial garden: an ideal model of sustainable development]. AB - There exist different kinds of views on the connotation of sustainable development of the society. In this paper, analysis from several aspects including certainty and necessity, the whole and the parts, development and limitation to the issue is given. Meanwhile, it can be shown from a summarized analysis to the models of sustainable development that the models consist of two kinds, theory one and practical one. A tentative plan of ecological industrial garden establishment in areas whose local conditions are permitted is put forward to promote its transformation rapidly from theory to practice, and it is regarded as a more perfect model for social sustainable development in the future. PMID- 11432053 TI - [Modeling research about bioremediation of organic pollutants in soil-water microbes system]. AB - A new Theory, organic pollutant sequestration inside soil particles, was applied in the research in order to explain the persistence of residual chemicals in remediation sites. Based on this theory, a mathematical model which simulates organic pollutant bioremediation process in soil-water-microbes system was developed. In the model, diffusion is represented by Fick's second law, reversible sorption-desorption by a linear isotherm, irreversible sequestration by a pseudo-first order kinetics, and biodegradation by Monod kinetics. Model results match successfully with experimental data. Model simulations are performed in the study. It is noteworthy that the mathematical model will be useful in quantitatively predicting the time and degradation extend of organic pollutant in remediation sites. PMID- 11432054 TI - [Fluoride emission from different soil minerals at high temperatures]. AB - The emission characteristics of fluoride pollutants from montmorillonite, kaolinite, vermiculite, geothite and allophane were studied to elucidate the mechanism of fluoride-releasing from soils during brick and tile making at high temperatures from 300 degrees C to 1000 degrees C. The rate of fluoride emission varied with temperature, mineral type, heating time, specific surface area and cations added to minerals. The escape of crystalline water resulting from crystal lattice collapse at a certain high temperature was found to affect the rate of fluoride emission. Calcium compounds could decrease fluoride emission rate from montmorillonite. At 800 degrees C, the rate of fluoride emission from Ca-treated montmorillonite decreased by 59.6% compared to untreated montmorillonite. The order for fluoride-fixing capacity of the 5 calcium compounds at 800 degrees C was as follows: CaCO3 > CaO > Ca3(PO4)2 > Ca(OH)2 > CaSO4. PMID- 11432055 TI - [Organochlorine residues in eggs of night heron breeding in Yuantouzhu, Wuxi and their functions as bioindicator]. AB - This study analyzed chlorinated pesticides in eggs of night heron breeding in Yuantouzhu, Tai lake. The HCH isomers, DDT, DDD, DDE, endrine and heptachlor epoxide were detected out in egg samples. The residue level and the percent of detection of beta-HCH and p,p'-DDE were significantly high. The mean value of p,p'-DDE was 0.906 microgram/g dry weight. Most of the chlorinated pesticides in samples from Wuxi were higher than those of Gongqing city, Jiangxi province. Oppositely, the hatching rate of night heron egg in Wuxi region was lower than that in Gongqing city. It may be related to the difference of DDE residual levels. Using organochlorine residue in eggs of water-fowls as an indicator for environmental quality assessment is feasible. PMID- 11432056 TI - [Modification of the Photobacterium phosphoreum toxicity test method]. AB - In order to reduce the variation of Photobacterium phosphoreum luminous intensity during testing, the Photobacterium phosphoreum toxicity test was modified by a correction factor, PIacetone, in this paper. The effects of magnetic stirring time, culture time, culture generation and culture temperature on the measured results were discussed. The modified method had good laboratory repeatability and the relative standard deviation (RSD) was between 2.1% and 13.1%. Reliable data could be obtained using the modified method. It would be beneficial to the development of the quantitative structure-activity relationships on multicomponent organic compounds. PMID- 11432057 TI - [Elevation distribution of precipitation chemistry in middle/upper troposphere in summer, the northern slope of central Himalayas]. AB - During 1997 summer, fresh snow samples were collected from four snowfall events from the elevation of 5800 to 7000 m in the Dasuopu Glacier region on the northern slope of Mt. Xixabangma (28 degrees 33'N, 85 degrees 44'E), in order to understand the elevation distribution of the middle/upper tropospheric components in the remote region. The correlation between SO4(2-), NO3-, Ca2+ and Mg2+ in precipitation differs with type of air mass, indicating the dominant sources of ions vary in short-term (e.g. several days) during summer low dust period. In general, with elevation increases, the concentrations of NO3-, Ca2+ and Mg2+ decreases, while SO4(2-) increases, which are influenced by the variation of dominant sources of ions. PMID- 11432058 TI - [A preliminary study on the organic carbon weathering fluxes in Beijiang River drainage]. AB - Riverine water samples were collected from Hekou hydrometric station of Beijiang River in five hydrological seasons. The samples were analyzed of their organic carbon and total suspended substance, which demonstrated that the concentrations of particulate organic carbon (POC) changed synchronically with the concentrations of total suspended substance (TSS) in riverine water. The correlation between the concentrations of TSS and dissolved organic carbon (DOC), however, is sometimes positive and sometimes negative, and the correlation is not so notable as that between TSS and POC. With the concentration of total suspended substances increasing, the quality partition of organic carbon in total suspended substance decreased in a logarithm tendency. The transportation of organic carbon, especially of DOC, mainly takes place in flood peak periods. The weathering flux of organic carbon in Beijiang River drainage basin is about 10.01 x 10(6) g.km-2.a-1, which is constituted by 6.54 x 10(6) g.km-2.a-1 of POC and 3.47 x 10(6) g.km-2.a-1 of DOC. This pattern of organic carbon weathering flux in Beijiang drainage basin is consistent with that of most of the monsoon drainage basins. PMID- 11432059 TI - [Chemical characteristics of fresh snow in Mount Everest Region]. AB - The chemistry of fresh snow samples collected in August and September, 1998 from Mount Everest was studied and compared with other fresh snow samples collected in different seasons in this region. The results indicated that major species in precipitation were very low in late summer in Mount Everest region and may be representative of the background of precipitation chemistry of remote regions in the wold. Chemical Characteristics of fresh snow in different seasons had distinct differences and they may reflect different moist source and climatic status. The precipitation in August and September was mainly come from Indian summer monsoon and in April and May it was influenced deeply by the dust of semi arid and arid regions in central Asia. This indicates the precipitation in this region is climatic sensitively. PMID- 11432060 TI - [Method on ozone generation with strong ionization discharge]. AB - This paper presents the formed methods of strong ionization discharge of dielectric barrier and plasma chemical reaction process of ozone generation. Ozone combination and decomposition are controlled by electric field intensity and electron energy. Therefore, new technologies with thinner dielectric layers (230 microns) of model alpha Al2O3 and narrow discharge gap (110 microns) are introduced, and strong ionization discharge is gained which reduced field (E) and electron average energy are more than 400Td and 10 eV respectively. Ozone concentration reaches to 200 g/m3 and ozone producing efficiency is 100 g/(kW.h). Ozone generator of big yield and miniaturization with module assembled method is realized. PMID- 11432061 TI - [Adsorption of p-chlorophenol by biofilm and its components]. AB - The p-chlorophenol (4-cp) adsorption to biofilm components was investigated by the experiment. Biofilm components included modeling water particulate (kaolin) with biofilm coating, bacterial cell, exopolysaccharide, kaolin and kaolin with exopolysaccharide. The adsorption of 4-cp in these systems could be described by Langmuir and Freundlich isotherm equations except the system of kaolin with exopolysaccharide. Under the condition of 25 degrees C and pH of 6.1 in these reaction systems, bacterial cell and EPS adsorbed more 4-cp than kaolin. The biofilm coating of kaolin affected its adsorption to 4-cp. Kaolin with biofilm coating adsorbed more 4-cp than that without biofilm coating. PMID- 11432062 TI - [The effects of iron compounds on SO2 and NO emissions in coal combustion process]. AB - The effects of ferric/ferrous chlorides and ferric oxide on emissions in Tongchuan coal combustion process were experimentally investigated using thermogravimetric analyzer. The results show that all iron compounds used in the experiments, namely FeCl3, FeCl2 and Fe2O3 can change the emissions of SO2 and NO, but with different mechanisms and different magnitudes. FeCl3 plays the roles of both a catalyst and an absorbent in the conversions from coal-S to SO2 and from coal-N to NO. Due to the catalysis the activation energy required for the formation of SO2 and NO is reduced, the rates of the reactions leading to SO2 and NO increased. The catalysis is concerned with the forms of coal sulfur. Due to the absorption FeCl3 reacts with SO2 to produce FeSO4 [symbol: see text] Fe2(SO4)3. In addition FeCl3 can decrease the emissions of CO, i.e., the combustion characteristics can be changed. The effects of FeCl2 on the emissions of SO2, NO and CO are similar to those of FeCl3, but weaker. Fe2O3 can also decrease the SO2 and NO emissions to some extent, but has negligible influence on CO emission. PMID- 11432063 TI - [Biological removal of VOCs emissions]. AB - The designed test conditions were following: vacancy velocity was from 0.7 cm/s to 3.5 cm/s, retention time was from 30 s to 80 s. Column activated carbon was selected as the filler of the reactor and toluene was taken as representative of VOCs. The biodegradation of toluene using biofilter reactor and the influence of concentration, flow and humidity on the toluene removal were investigated and the removal model was set up. The experimental results showed that the biofilter reactor had higher toluene removing ability. The observation of biotic community demonstrated that the microbes consisted of fungi, bacillus and spore bacillus. Of them spore bacillus was dominant. PMID- 11432064 TI - [Computer simulation of HA influence on copper bioavailability to the fish gills]. AB - Copper speciation in the gill microenvironment of neon terris (Paracheirodon innesi) and the influence of humic acid and pH on copper bioavailability were investigated using exposure experiment and chemical equilibrium calculation. It was found that the copper speciation was changed because of the influence of the humic acid. With the existence of humic acid, the dominant species of copper in the bulk solution was humic acid complexed copper. However, humic acid complexed copper along with mucus complexed copper dominated the fish gill microenvironment. Consequently, the bioavailable copper species both in the bulk solution and in the fish gill microenvironment were significantly reduced by humic acid. The result of the modeling also indicated that the influence of pH was more significant under alkaline condition. PMID- 11432065 TI - [Biodegradation behavior of ethylenediaminetetraethylene acid]. AB - Aerobic and anaerobic degradability of ethylenediaminetetraethylene acid(EDTA) were measured on Warburg respirometer and anaerobic batch reactors. Biodegradation of EDTA as a single substance was poor, but biodegradation of EDTA with co-substrate could be improved. The wastewater containing EDTA can be treated by activated sludge process. The EDTA removal efficiency was 80% under the circumstance HRT 16 h and EDTA removal efficiency reached to 92.5%-95.1% when HRT was 20 h. The main factor affecting the biodegradability of EDTA is sludge remained time. PMID- 11432066 TI - [Extinction characteristics of TiO2 suspension for different band ultraviolet radiation]. AB - This study focused on the global absorption characteristics of UVA, UVB and UVC radiation in the suspension of titanium-pollutant solution using 4-CBA-Na as a model pollutant. The result showed that the attenuation of ultraviolet in the suspensions conformed to Beer's Law, light intensity negative-exponentially attenuating with the optic path. The extinction coefficient increased with the titanium dosage by a quadratic polynomial law, which may be approximated by a linear function at lower dosage. The sum of the respective UVC or UVB extinction coefficient in pollutant solution and titanium-water suspension was found to approach the measured value in Titanium-Pollutant suspension. Larger extinction coefficient was observed for shorter UV wavelength, implying higher energy efficiency with UVC as ultraviolet source. PMID- 11432067 TI - [Single particle source apportionment of atmospheric aerosol based on proton microprobe]. AB - A proton microprobe with high resolution and high sensitivity was applied to analyze atmospheric aerosol in the people's park in Shanghai urban for single particle source apportionment. The results showed that the sum of the average contribution rates from the sources of soil dust, building construction dust, vehicle exhaust and metallurgical industry, which are the main pollution sources of atmospheric aerosol in Shanghai urban and surpasses 80%. Besides, about 13% of the particles could not be identified. On the basis of cluster analysis of these particles, they are preliminary known belonging to two different individual sub pollution sources and a new unidentified pollution source. PMID- 11432068 TI - [A study on city motor vehicle emission factors by tunnel test]. AB - Applying the principle of tunnel test to run a typical across-river tunnel test in Guangzhou city, 48 h-online-monitor data include pollutant concentration, traffic activity and meteorological data were gained. The average motor vehicle emission factors of NOx, CO, SO2, PM10 and HC were calculated using mass balance which are 1.379, 15.404, 0.142, 0.637, 1.857 g/km. vehicle respectively. Based on that, combined emission factors of 8 types of city vehicles were calculated using linear regression. The result basically showed the character and level of motor vehicle emission in Chinese city. PMID- 11432069 TI - [Atmospheric particulate mercury concentration in Changchun City and its dry deposition flux]. AB - From July 1999 to January 2000, the particulate mercury(Hg(p)) concentration of five function sites and one contrast site were determined. In non-heating time, the particulate mercury concentration in urban atmosphere ranged from 0.022 to 0.398 ng.m-3 with an average of 0.145 ng.m-3 and the contrast site with an average of 0.084 ng.m-3. In heating time, the Hg(p) concentration in urban was in the range of 0.148-1.984 ng.m-3 with an average of 0.461 ng.m-3 and with an average of 0.211 ng.m-3 in the contrast site. The Hg(p) concentration in heating time was two times higher than that of non-heating time. Coal fire and wind-blown soil material were the two important sources of Hg(p). Wind-blown soil material contributed 7.9%-38.5% of the Hg(p) with an average of 24.1% in urban. Use theory model to estimate the dry deposition flux with the results of 43.06 micrograms.(m2.a)-1 in urban and 21.28 micrograms.(m2.a)-1 in the contrast site. PMID- 11432070 TI - [Dust deposition and its measurement by 137Cs in the Gonghe Basin, Qinghai Province, China]. AB - Through the dust observation at three sites in the Gonghe Basin, Qinghai Province, China and 137Cs analysis of dust samples, the 137Cs technique is firstly used to study the dust deposition by storm. The rate of dust deposition and its monthly distribution were determined and the dust source traced by 137Cs. It is apparent that the 137Cs model for accumulation rate must be modified because of the 137Cs redistribution from dust gain. These tentative results may be have some significant implications for further dust storm research. PMID- 11432071 TI - [Multimedia environmental behavior of herbicide atrazine]. AB - In the paper, taking the Baiyangdiang Lake area as a study field, on the basis of soil characterization, environmental monitoring, and studies of adsorption, biodegradation, hydrolysis and photolysis, the atrazine concentrations in soil, groundwater and corn plant during 30 years were predicted by multimedia environmental fugacity model. PMID- 11432072 TI - [Comprehensive assessment on environmental quality in vulnerable ecotone of Tarim River Basin]. AB - Tarim River Basin (TRB) is one of ecological environment vulnerable areas. Due to the matching of material and energy existed in some problems, the different fragility characteristics occurred obviously. According to related principles of ecological environment quality assessment, combined with ecological environment situation of Tarim River Basin, 20 indexes were selected, and the indicator system for synthetically ecological environment assessment was built. Moreover, ecological fragility index was set up, which can be used to indicate the degree of ecological environment quality in Tarim River Basin. The results were as follows: Aksu River Basin belongs to improved area. Yerkart River Basin and the upper reaches of TRB belongs to balance areas and Hetian River Basin and the middle reaches of TRB belongs to maladjustment areas. The lower reaches of TRB belongs to severe damage area. The assessment results are consonant with actual situation, which play an important function in ecological environment construction. PMID- 11432073 TI - [The absorbed rate and distribution rule of rare earth elements in wheat by 141Ce as tracer]. AB - The study on absorption, distribution rules and transportation rates of cerium in the interface of water-wheat by 141Ce in culture solution were carried out. The results showed that the concentrations of cerium in wheat seedling were gradually increased during the experiment, and so did the same after increasing the concentration of cerium adding in the culture solution. The concentrations of cerium in roots of winter wheat were significantly high than that in tops (steam & leave). The absorption rates of cerium in wheat with different treatments of cerium were distinct. The quantitative formulas, which describe the rule of absorption rate and culture time, was given resulting from the fit of the experimental data. PMID- 11432074 TI - [The efficiency of algae removal from drinking water by ferrate]. AB - This paper studied the efficiency of algae removal by ferrate from two kinds of raw water which were collected from Donghu and Tiegang reservoir respectively. The experimental results showed that the removal efficiency of algae reached about 95% only through PAC coagulation when the raw water was extracted from Donghu reservoir in which most of algae was Chlorella. But dealing with Tiegang raw water in which the majority of algae were Oscillatoria, terrate was used as pre-oxidant coupling with PAC coagulation not only using PAC to flocculate and the efficiency of algae removal could be up to 97.85%. However this treatment method using ferrate as pre-oxidant overmatched the traditional method prechlorination for algae removal. PMID- 11432075 TI - [Anaerobic biological treatment of Lincomycin production wastewater]. AB - The high-strength Lincomycin production wastewater containing toxic and refractory substances treated by lab-scale mesophilic UASB reactor was described. When the reactor was operated in influent COD 8000-14,000 mg/L and HRT 10 h, the volumetric loading rate and COD removal rate could reach 20-35 kg/(m3.d) and 50% 55%, respectively. The granular sludge might be formatted by using a bit longer acclimation time, adjusting and maintaining fairly high surface hydraulic loading rate of 0.2-0.4 m3/(m2.h), influent COD of 2000-3000 mg/L and sludge loading rate of 0.2-0.5 kg/(kg.d). The anaerobic kinetic constants of Vmax and Ks for the wastewater treatment were 1.3 d-1 and 8133 mg/L, respectively. The non biodegradable substances accounted for about 30% of total COD, which was the important factor of relative low COD removal rate for the wastewater. PMID- 11432076 TI - [Aerobic sludge granulation and biological phosphorus removal in different operating conditions of SBR]. AB - This paper investigate the effect of the different operating conditions on the aerobic sludge granulation and biological P removal characteristics in SBR by using synthetic domestic wastewater as feed. Different ratios of COD/TN (11.79, 18.85, 20.72, 24.66), COD/TP (27.85, 44.53, 48.93, 58.25), TN/TP (4.00, 2.36, 1.54), temperature(22 degrees C, 15 degrees C, 8 degrees C) and sludge retention time(16d, 10d, 5d) are tested in this process. It is found that the high ratios of COD/TN (24.66), COD/TP (58.25), temperature (22 degrees C) and low SRT (10d) are beneficial to the phosphorus removal and the cultivation of the aerobic granular sludge. The proper TN/TP rate (2.36), dissolved oxygen and selection of anaerobic seed sludge are basic requirement and most important factors to guarantee such aerobic granules in the process. PMID- 11432077 TI - [Photocatalytic degradation of bromomethane on sol-gel-derived SO4(2-)/TiO2]. AB - SO4(2-)/TiO2 photocatalysts were prepared by sol-gel method and characterized by X-ray diffraction (XRD), FTIR spectroscopy and N2 sorption analysis applying BET method. Photocatalytic decomposition of CH3Br in air stream was studied over SO4(2-)/TiO2 photocatalysts under the different reaction conditions. The results showed that the addition of SO4(2-) to TiO2 greatly enhanced its structural and photocatalytic properties. The optimum concentration of SO4(2-) on TiO2 was 9% wt and the best calcination temperature was 450 degrees C. Moreover, the sulfated TiO2(SO4(2-)/TiO2) exhibited excellent stability of activity under the moist reaction condition. When the reaction temperature was below 85 degrees C, the reactivity of bromomethane was enhanced with increasing reaction temperatures, and the apparent Arrehenius activation energy was 19.6 kJ.mol-1. In the reaction temperature range of 85 degrees C-105 degrees C, the apparent Arrehenius activation energy was zero. PMID- 11432078 TI - [Experimental and mechanism analyses on HCl emission control during PVC combustion in fixed beds]. AB - Incineration of chloridated plastic (PVC) in solid wastes contributes much to HCl pollutant in air environment. The characteristics of HCl emissions and reduction during PVC combustion were conducted in a nearly isothermal, externally heated, horizontal quartz-tube reactor. With increasing temperature from 700 to 900 degrees C, the conversion of HCl increased leniently from 82.5% to 88.3%. High excess air factor enhanced a small amount of HCl existing in form of Cl2. The calcium-based sorbents including CaCO3, Ca(OH)2, Ca(CH3COO)2 exhibited high HCl capture efficiencies from 68% to 79%, while magnesium-based sorbent less than that of 3%. Influences of operational variables, i.e., size of sorbent, Ca/Cl molar ratio and combustion fume compositions, on the HCl reduction were discussed. From a point of view of equilibrium constant of dechloridization reaction the experimental results were explained in detail. PMID- 11432079 TI - DOs question intent of challenge. PMID- 11432080 TI - Osteopathic medical schools should foster sense of identity. PMID- 11432081 TI - Without sampling information, data lack impact. PMID- 11432082 TI - New cholesterol guidelines for the new millennium. PMID- 11432083 TI - Oral polymeric N-acetyl-D-glucosamine and osteoarthritis. AB - Many patients with arthritis are using alternative modes of therapy, including nutritional supplements, to treat their arthritis. Most patients never tell their doctors that they are taking alternative medications, and few doctors even ask about such activities. Over-the-counter supplements are expensive and consume large amounts of patients' healthcare dollars. Glucosamine has been widely touted as being an effective arthritis treatment. The authors designed and undertook a study to test the efficacy of a polymer of N-acetyl-D-glucosamine (NAG), or POLY Nag, in a double-blind, placebo-controlled study in patients with osteoarthritis. Results indicate that POLY-Nag may be useful in treating patients with osteoarthritis. PMID- 11432084 TI - The primary care physician's role in caring for internationally adopted children. AB - The rate of international adoption in the United States has been increasing. Internationally. adopted children have various medical, psychological, and developmental issues that need to be evaluated and addressed by primary care physicians. Full histories and physical examinations, developmental assessments, and screening tests are necessary for such children. Family concerns and cultural differences between the child and the adoptive family need to be addressed as well. PMID- 11432085 TI - Correlation of scores for the Comprehensive Osteopathic Medical Licensing Examination with osteopathic medical school grades. AB - The authors evaluated construct validity of scores for the Comprehensive Osteopathic Medical Licensing Examination (COMLEX-USA), the examination used to evaluate osteopathic physicians for licensure. They computed correlations between students' grades in the first 2 years of osteopathic medical school and their scores on the COMPLEX-USA Level 1 (N = 187) and Level 2 (N = 86), as well as correlations between third- and fourth-year clerkship grades and the COMLEX-USA Level 2. Correlations of Level 1 scores with grades for years one, two, and the first 2 years combined were .74, .80, and .81, respectively; for Level 2, correlations were .59, .70, and .71. Correlation between clerkship grades and scores for the COMLEX-USA Level 2 was .26. The strong correlation between COMLEX USA results and grades for the didactic curriculum in the first 2 years of medical school provides evidence for the construct validity of scores for the COMLEX-USA Levels 1 and 2. PMID- 11432086 TI - "The California incident". PMID- 11432087 TI - [Utility of echo enhancers in the study of pulmonary circulation time]. AB - Pulmonary circulation time (PCT) varies under different clinical conditions. To evaluate the PCT, the persistence of contrast in both ventricles and the digital arterial saturation of oxygen with peripheral intravenous injection of microbubbles, were examined 14 patients. Groups: I normals, II left ventricular hypertrophy (EF > 50%), III with mitral stenosis and IV in congestive heart failure. In seven patients, isosorbide S/L was administered after the first injection. The transpulmonary transit in beats was 7.3 in normals, and larger in the others. The persistence of echo contrast in right chambers was 31 beats in group I, and larger in the other groups. In left chambers it was 20.6 beats in group I, and larger in others. There were no alterations with regards to peripheral arterial saturation of oxygen. AII patients with isosorbide presented shorter PCT and persistence of contrast in right chambers. CONCLUSIONS: There are differences in PCT among normals, patients with mitral stenosis and those with congestive heart failure. Prolonged persistence contrast in right chambers indicates abnormally slow transpulmonary transit. Patients with congestive heart failure and ventricular damage, have longer persistence of echo contrast. PCT studied with contrast is an easy means of evaluating some aspects of pulmonary circulation. PMID- 11432088 TI - [Trends for bacteremia and risk factors for death in a tertiary hospital in Mexico City. 1981-1992]. AB - OBJECTIVE: To describe the trends and risk factors of death for bacteremia in adult from a tertiary-care center from 1981 to 1992. MATERIAL AND METHODS: We randomly included 20% of bacteremic episodes per year. RESULTS: 47,618 blood cultures from 19,530 patients, 3428 patients (17.6%) had bacteremia (285/y). From 600 episodes (50/y), 307 were from men, 368 were hospital-acquired (HA), and 88% were monomicrobial. Diabetes mellitus was seen in 103 cases, cirrhosis of the liver in 98, and AIDS in 33, among others. The main microorganisms were: Escherichia coli (177), Klebsiella pneumoniae (53), Enterobacter (50), Salmonella (45) and Pseudomonas aeruginosa (35); coagulase-negative staphylococci (CNS) (116), Staphylococcus aureus (56), and enterococci (22), and Candida (20). CNS decreased during the study (p < 0.01), but Candida spp., Stenotrophomonas maltophilia and enterococci increased (p < 0.01). The crude mortality of the HA bacteremia was 70.8%, and 29.2% in the case of community-acquired, the mortality attributable to HA bacteremia was 41.6%. The main risk factors were: cardiac valvular disease (p < 0.001), stay at the intensive-care unit (p < 0.001), sepsis (p < 0.001), and pneumonia (p < 0.001). DISCUSSION: Bacteremia had a significant impact on mortality during the study period that has not change despite opportune therapy, Enterococci and candida have emerged as significant pathogens. PMID- 11432089 TI - [Safety and efficiency of Echogen. Transthoracic echocardiography study]. AB - This study was performed to determine the safety and efficacy of intravenous dodecafluoropentane emulsion (Echo Gen), to determine the efficacy and duration of contrast agent in left cavities by transthoracic echocardiography and to evaluate the clinical impact of Echogen. Fifteen patients were studied, by transthoracic echocardiography. Duration of contrast intensity of left ventricular cavity opacification and endocardial border definition were studied. The adverse effects were assessed immediately, 24 hours and 7 days after Echogen. Both the vital signs and electrocardiogram did not show any significant changes. Minimal changes in hepatic function in one patient and in renal function in other one were observed. The mean duration of left ventricular opacification was 4 minutes 47 seconds and full or intermediate opacification was more frequently observed after Echogen. Endocardial border delineation was poor in one patient, intermediate in five and excellent in nine. In 86.7% the diagnostic information obtained was sufficiently and allowed other studies. CONCLUSIONS: This study demonstrates that Echogen is effective for Left ventricular cavity opacification, endocardial border definition and assessment of left ventricular volume and ejection fraction 2) The obtained information with the use of Echogen allowed other studies. 3) The intravenous administration of Echogen is safe in doses of 0.05 ml/Kg. PMID- 11432090 TI - [Risk factors for myringoplasty failure]. AB - OBJECTIVE: Identify risk factors associated with failures in myringoplasty. MATERIAL AND METHODS: 290 cases were reviewed in the period of july 1997 to march 1999. An analysis of cases and controls was performed. RESULTS: The 290 myringoplasties were analyzed. The minimum period of follow-up was: 6 months. The estimated surgical success was 82.1%, and 17.9% of the cases were considered as a failure. There were no risk factors found statistically significant to the variables: age, sex, schooling or occupation of the patient, tobacco usage, alcoholism and associated illnesses; prior nasal or otological surgery or cause of the perforation nor evolution time whether or not it was a resident doctor who performed the surgery. These results are not definitive since the intervals of confidence were ample in the majority of these variables, which can be attributed to the size of the sample group. The significant risk variables found were: size of perforation greater than 40%, marginal localization of the perforation; tympanosclerosis, scar tissue and cholesteatoma. CONCLUSIONS: The risk factors in the myringoplasty failure are similar to those reported in diverse studies. The variables highly significant for failure are details that should be watched in the technique. PMID- 11432091 TI - [Pharmacogenetics and its importance in the clinical]. AB - Therapeutic inefficacy and pharmacologic toxicity has frequently been seen in the clinical use of drugs or medicines in individuals under pharmacological treatment. Due to the presence of drug metabolizing enzymes, medicines may participate as substrate inhibitors or enzyme inductors. Their activity may vary among individuals. This enzymatic variability may be assessed through the analysis of recombinant DNA, using restriction analysis of the genomic DNA (fragment restriction of polymorphic length) and the enzymatic amplification of DNA through PCR (polymerase chain reaction). This technology has been used in clinical studies which allow us to know the mechanisms of inherited variations in response to drugs regulated by each individual's genes dependent on different races. These enzymatic differences may also be influenced by nutritional habits or environmental factors. This study deals with the importance of understanding the metabolism of drugs applied to the therapeutic management of individuals with therapeutic inefficacy or pharmacologic toxicity. PMID- 11432092 TI - [Equine estrogens vs. esterified estrogens in the climacteric and menopause. The controversy arrives in Mexico]. AB - It exists controversies about if the effects and benefits of the esterified estrogens could be similar to those informed for equines, because its chemical composition and bioavailability are different. Esterified estrogens has not delta 8,9 dehydroestrone, and its absorption and level of maximum plasmatic concentrations are reached very fast. In United States of America and another countries, esterified estrogens has been marketed and using for treatment of climacteric syndrome and prevention of postmenopausal osteoporosis, based on the pharmacopoiea of that country, but the Food and Drug administration (FDA) has not yet authorized up today, a generic version of conjugated estrogens. In Instituto Mexicano del Seguro Social (IMSS) and another institutions of health sector in Mexico, starting in year 2000, it has been used esterified estrogens for medical treatment of climacteric and menopausal conditions. For this reason, in this paper we revised the most recent information about pharmacology, chemical composition, clinical use and costs of the conjugated estrogens with the purpose to guide the decisions to purchase this kind of drugs in Mexican heath institutions. PMID- 11432093 TI - [78-yr-old male patient with dementia]. PMID- 11432094 TI - [Severe hypoglycemia secondary to angiotensin-converting-enzyme inhibitors in the absence of diabetes mellitus. Report of a case]. AB - We present the first reported case of untreatable hypoglycemia in a woman of 66 years-old, non-diabetic who received captopril for six years, and more recently ramipril. Six months ago, she was seen for the first time in our hospital. We found her disoriented, sweaty and confusing words frequently. Glucose tolerance test one hour after administration of glucose: 53 mg/dl. Test of insulin and C peptide had norms the same as the other contra-regulator hormones except glucagon with moderate elevation. The B.P. increased and we changed to ramipril, which improved the pressure but the hypoglycemia persisted. A new change was made to Losartan plus hydrochlorothiazide; after 72 hours patient improved. A new glucose tolerance test one hour after administration of glucose is absolutely normal. General conditions of the patients are observed more than 3 months and are asymptomatic. The B. P. is normal and we are keeping only the recently installed treatment. PMID- 11432095 TI - [Mechanical mitral valve prosthesis dysfunction in pregnant women. Report of 2 cases]. AB - We present two cases of pregnant women associated with mechanical mitral valve prostheses dysfunction, treated between October 1998 and September 2000, in congestive heart failure, functional class III, without fetal distress. Emergency surgery consisted in cesarean procedure immediately followed for mitral prostheses removal and implantation of new mechanical heart valve prostheses was realized in both patients. In each case a single live infant was obtained with good Apgar evaluation. Both patients had successful surgical procedures. In both patients were found thrombi in the mechanical heart valves. Actually patients and their children have good evolution, without any problem. PMID- 11432096 TI - [Physicians and medicine in 16th century New Spain]. AB - The more prominent physicians and surgeons, European, native and creole, who practiced their art in New Spain during the XVI century, are remembered. There were improvised surgeons among the Spanish soldiers, who faced the American natives in the name of universal empire and church. There were also native physicians, organized around an important cultural center: the Franciscan college of Holy Cross in Tlatelolco. They perpetuated the ancestral medical traditions. In the dawning of New Spain, arrived here some physicians and surgeons prepared in important medical centers, such Sevilla, Salamanca, and Alcala de Henares. Soon after a noteworthy exchange of medicinal plants and, generally, of therapeutic products between the old and new world took place. Likewise arrived here medical books printed in Europe and, in the second half of such century, appeared Newspanish medical books. When the first chair of medicine was established in the Royal University of Mexico (1578), the number of medical publications increased until, in 1598, appeared the first medical thesis printed in America. PMID- 11432097 TI - [Laparoscopic diagnosis of a simple hepatic cyst]. PMID- 11432098 TI - [The Human Genome sequence]. PMID- 11432099 TI - [Bioethics in the face of death]. AB - We review death, thanatology and bioethics concepts and precepts, the value scale and hierarchization; the changes in death vision according to culture, religion and hierarchy, changes in perception of, according to culture, religion and mores in different communities and times, as well with scientific and technological advances. We analyzed patient's reactions to death, and the reactions of people close to them. We describe and analyze the principal bioethical dilemmas associated with death: therapeutic overkill or dysthanasia, passive and active euthanasia, assisted suicide, orthothanasia, and organ transplants. We discuss the relationship between death and science, bioethics and thanatology, as a necessary discipline today. PMID- 11432100 TI - [Legal update on medical practice]. PMID- 11432101 TI - [The democratization of health. A vision for the future of the health system in Mexico]]. PMID- 11432102 TI - Presidential address. Strands of history. PMID- 11432103 TI - Distraction osteogenesis and current bone grafting techniques. PMID- 11432104 TI - Transmyocardial revascularization: from theory to practice and back again. PMID- 11432105 TI - A clinical decision to save the tooth or place the implant. PMID- 11432106 TI - Microscopic surgery. PMID- 11432107 TI - Ceramics. PMID- 11432108 TI - Effect of duration of estradiol replacement on the outcome of oocyte donation. AB - PURPOSE: To investigate if duration of estrogenic endometrial stimulation can affect recipient pregnancy rate in an ovum donation program. METHODS: Each recipient received micronized 17 beta-estradiol orally in a steadily increasing dosage from 2 to 6 mg daily over a period of time varying from 5 to 76 days until oocyte were available for donation. Recipients (520 patients for a total of 835 transfer cycles) were retrospectively divided into five groups depending on the duration of E2 administration. RESULTS: No significant difference was seen in pregnancy and implantation rates between groups. There was a higher number of miscarriages in Group A (41%), p < 0.05 vs. Group B (15%), and vs. Group E (1%). Age, number of pregnancies and miscarriages, or implantation rate in donors (327 women aged < 35 years) were similar in all the five groups. CONCLUSIONS: Endometrial receptivity is tolerant to a wide duration of E2 treatment (until 2 months), while waiting for oocytes available for donation, but best results are achieved with a treatment range of 11 to about 40 days. PMID- 11432109 TI - Comparison of recombinant and urinary follicle-stimulating hormone preparations in short-term gonadotropin releasing hormone agonist protocol for in vitro fertilization-embryo transfer. AB - PURPOSE: To compare the efficiency and efficacy of two recombinant human FSH (r FSH) and urinary (u-FSH) preparations in patients undergoing superovulation for IVF-ET using a short-term gonadotropin releasing hormone agonist (GnRH-a) (Triptorelin) protocol. METHODS: A total of 88 women undergoing IVF-ET were included in this prospective study. They were randomized to receive u-FSH (150 IU/d), follitropin-alpha (100 IU/d), or follitropin-beta (100 IU/d) for 2 days, and dosages were subsequently adjusted according to the ovarian response. RESULTS: The FSH dose required for the overall stimulation was significantly lower in patients treated with r-FSH than in those treated with u-FSH while serum FSH values were higher in the latter group. There were no statistically significant differences in ovarian response and IVF outcome between r-FSH preparations. CONCLUSIONS: Recombinant FSH preparations have a higher efficiency than urinary ones in patients undergoing IVF-ET using a short-term GnRH-a protocol. In this situation, the two recombinant follitropins have comparable effectiveness. PMID- 11432110 TI - Improvement of IVF outcome in poor responders by discontinuation of GnRH analogue during the gonadotropin stimulation phase--a function of improved embryo quality. AB - PURPOSE: To assess the efficacy of a protocol involving the discontinuation of the GnRH analogue at the mid-phase of ovarian stimulation for IVF in patients with a previous poor response. METHODS: Prospective case-control evaluation compared with same patient's previous performance. Thirty-six patients enrolled in an IVF program were treated in two consecutive cycles. The first with a standardized protocol utilizing midluteal administration of Nafarelin (N) 600 mcg/d continued throughout the stimulation phase with human menopausal gonadotropin (hMG) until follicles of 20 mm were identified by transvaginal ultrasound (Standard group). Patients with a poor response in the Standard cycle were treated in the subsequent cycle with N and hMG initially in a similar manner, then N was stopped after 5 days of hMG stimulation (N-stop group). All clinical and laboratory aspects of treatment were done in a similar fashion in both cycles, each patient acting as her own control. RESULTS: Results were analyzed by paired t test. The change in each parameter in the N-stop cycle was expressed as the percent change as compared with the standard protocol cycle for each patient. Peak estradiol (E2) and number of aspirated oocytes were increased in the N-stop cycle (+16.9% and +28%, respectively), but insignificantly so. The percent of cleaving embryos was significantly increased by 27.9% (p = 0.03) in the N-stop cycle, as embryo morphology was improved by 22% (p = 0.02). The efficacy of gonadotropin treatment was enhanced in the N-stop cycle, as expressed by a 32.5% increase in oocytes retrieved per hMG ampoule administered (p = 0.04). Three cycles of 36 were cancelled during the N-stop cycle, whereas only one was cancelled in the standard protocol cycle. Of the 36 patients, 7 conceived in the N-stop protocol and 5 are ongoing pregnancies. CONCLUSION: Discontinuation of GnRH-a during ovarian stimulation for IVF has a beneficial, but not statistically significant, effect on both E2 and oocyte production. Embryo cleavage rates and morphology were significantly improved, this may be due to improved oocyte quality, which may have been responsible for achieving pregnancies. The efficacy of gonadotropin treatment was enhanced when GnRH-a was discontinued. These results hint that GnRH-a may have a direct negative effect on folliculogenesis and oocytes, which is apparent especially in poor responder patients. PMID- 11432111 TI - IVF-patients with nonmale factor "to ICSI" or "not to ICSI" that is the question? AB - PURPOSE: Intracytoplasmic sperm injection (ICSI) guarantees high fertilization rates and could theoretically lead to higher implantation rates as well. Furthermore injection into oocyte creates a hole in the zona pellucida similar to the procedure of assisted hatching. We were therefore interested to assess such a potential benefit for infertile IVF patients without male factor. MATERIALS AND METHODS: Open randomized prospective study according to the rules "Good Clinical Practice" with informed consent of the patients and institutional review board approval. Ninety-one consecutively seen patients with tubal infertility or hostile cervical mucus were randomized to undergo either ICSI (44 patients) or IVF (45 patients). In two patients fertilization of oocytes failed and so a repeated ICSI had to be performed. All these patients were stimulated with the same protocol, using the gonadotropin releasing hormone-agonist (GnRH-a) buserelin acetate in an ultrashort flair-up protocol together with pure follicle stimulating hormone (rFSH). The two study groups did not differ in terms of age, BMI, and all baseline hormone levels. RESULTS: The total pregnancy rate was 42% in the normal IVF group with 33% ongoing pregnancies. The ICSI group had a total pregnancy rate of 39% with 23% ongoing pregnancies. The implantation rate per transferred embryo was higher for normal IVF but not significant (18% versus 11%). The variables, fertilization rate, age, body mass index, baseline hormone levels, endometrial thickness, embryo score, and the highest grade embryo per transfer were very similar in both groups. CONCLUSION: ICSI should be applied only when conventional IVF fails, that is, for male factor patients and for patients with unexplained infertility. PMID- 11432112 TI - Embryo selection by the first cleavage parameter between 25 and 27 hours after ICSI. AB - PURPOSE: The objective of this study was to assess first embryo cleavage (FEC) 25 27 h after intracytoplasmic sperm injection (ICSI) as a parameter for the embryo selection process. METHODS: From January 1998 to December 1999, a total of 670 patients were subjected to the ICSI programme at the Centre for Human Reproduction, Sinha Junqueira Maternity Foundation, and the FEC parameter was evaluated in three situations. RESULTS: In the first, a total of 300 zygotes were analyzed on the basis of a score (16-18 h after ICSI) and observed for the presence or absence of FEC (25-27 h after ICSI). A significant (p < 0.02) presence of FEC was observed in zygotes with a score of 15 (ideal score). In the second, a total of 200 patients were selected and divided into two groups matched for age and laboratory performance. Group I (n = 100) was subjected to transfer of embryos with the absence of FEC only (since in this cycle no embryos with FEC were detected within 25-27 h after ICSI) and Group II (n = 100) was subjected to transfer of embryos with the presence of FEC only. The age of Group I patients (33.8 +/- 4.2 years) did not differ significantly (p = 0.50) from that of Group II patients (33.5 +/- 4.3 years). The number of embryos transferred was similar (p = 0.07) for Group I (2.7 +/- 1.1) and Group II (2.9 +/- 0.88). In Group II, the 17.5% implantation rate was significantly higher (p < 0.01) than the 5.9% rate obtained for Group I. The pregnancy rate for Group II was significantly higher (p < 0.01) (33%) than that for Group I (12%). The incidence of abortion was 16.6% in Group I as compared with 6% in Group II. In the third situation, we observed the frequency of embryos with FEC in 36 patients whose implantation rate was 100% (ideal result) and obtained a value of 82%. CONCLUSIONS: The data suggest that the presence of the FEC parameter that was evaluated 25-27 h after ICSI could be used to select embryos with a higher implantation power. The data reported here may justify routine analysis of embryos with FEC for the process of embryo selection after ICSI. PMID- 11432113 TI - Relationships between sperm motility characteristics assessed by the computer aided sperm analysis (CASA) and fertilization rates in vitro. AB - PURPOSE: Some studies have suggested that computer-aided sperm analysis (CASA) estimates of concentration and movement characteristics of progressively motile spermatozoa are related to fertilization rates in vitro. However, it has also been suggested that the greater number of motility parameters assessed by CASA does not imply more precision in predicting fertility. This study was carried out to investigate the relationships between the CASA estimates and fertilization rates in vitro. METHODS: Semen quality analysis was performed using CASA in 136 in vitro fertilization-embryo transfer (IVF-ET) cycles with at least 3 oocytes collected. The CASA estimates before and after swim-up were compared between 108 cycles with fertilization rate > 50% ("good" group) and 28 cycles with fertilization rate < or = 50% ("poor" group). RESULTS: Before swim-up, there were significant correlations between fertilization rates and CASA estimates, including amplitude of lateral head displacement (ALH) (r = .269), curvilinear velocity (VCL) (r = .297), straight line velocity (VSL) (r = .266), and rapid sprm movement (Rapid) (r = .243). There was also a significant correlation between the fertilization rates and straightness (STR) after swim-up (r = 0.178). As for sperm movement characteristics, there were significant differences of ALH (p < .005), VCL (p < .001), VSL (p < .005), and Rapid (p < .01) between "good" and "poor" groups before swim-up. After swim-up, there were significant differences of VCL (p < .005), average path velocity (VAP) (p < .005), and Rapid (p < .05) between the two groups. CONCLUSIONS: These results indicate that some of the CASA estimates provide reliable estimation of the fertilizing ability of human sperm. There were significant differences of the two sperm movement characteristics, including VCL and Rapid (before and after swim-up), indicating that the total distance traveled by rapid sperm movement might be important in human sperm fertilizing abilities. PMID- 11432114 TI - Relation between different human sperm nuclear maturity tests and in vitro fertilization. AB - PURPOSE: To consider the relationship between different sperm nuclear maturity tests and in vitro fertilization (IVF) rate, in order to select the most sensitive, specific, and independent factor(s) for prediction of in vitro fertilization. METHODS: Infertile couples (101) were randomly selected from IVF candidates referred to Isfahan Fertility and Infertility center. Semen samples were collected on the day of oocyte recovery. Following routine semen analysis, major portion of the semen was prepared for routine IVF insemination and the remaining was used for following sperm nuclear maturity tests: chromomycin A3 (CMA3), aniline blue, sodium dodecyl sulphate (SDS) test, and acridine orange test with or without heat shock (87 degrees C, 5 min). Sperms (200) were evaluated for each test. The results were recorded and analyzed for their correlation to fertilization rate, using correlation coefficient, logistic regression analysis, student t-test, and receiver operating characteristics (ROC) curve. RESULTS: Among these tests, aniline blue and CMA3, and semen parameters, sperm morphology, and sperm motility showed a significant correlation with fertilization rate. Using logistic regression analysis, sperm morphology and CMA3 were the only independent factors related to in vitro fertilization. ROC curves showed that among above tests, CMA3 is the most specific and sensitive for sperm nuclear maturity. CONCLUSION: Among CMA3, aniline blue, SDS test, and acridine orange, CMA3 was the most sensitive and specific test that can be used along with routine semen analysis for more precise prediction of fertilization rate. PMID- 11432115 TI - Triple-test screening in in vitro fertilization pregnancies. AB - PURPOSE: To determine whether the results of triple-test analysis differ between spontaneous and IVF pregnancies. METHODS: The study population consisted of 140 women with singleton pregnancies, 70 by IVF, and 70 by spontaneous conception. The groups were matched for maternal age, gestational week, and laboratory batch. The levels of all triple-test markers--alpha-fetoprotein (AFP), human chorionic gonadotropin (HCG), and unconjugated estriol (u-E3)--were compared between the groups. RESULTS: Significantly higher HCG levels were detected in the patients with IVF pregnancies than in the control group (1.31 +/- 0.8 vs. 0.95 +/- 0.5 multiple of the medians, respectively, (p < 0.006), but there were no significant differences in AFP or u-E3 levels. Overall, 18.5% of the IVF group were found to be screen-positive as compared with 11.4% of the control group (difference not statistically significant). Only 8 IVF pregnancies (11.4%) reached the lowest calculated risk possible (1:9999) compared with 17 (24.2%) in the control group (p < 0.05). CONCLUSIONS: Our findings support previous data demonstrating elevated maternal serum HCG in IVF patients in comparison with spontaneous ones. PMID- 11432116 TI - Evaluation of the meiotic spindle apparatus in metaphase II human oocytes following cytoplasmic donation. AB - PURPOSE: To determine if the removal of cytoplasm from metaphase II human donor oocytes damages the meiotic spindle apparatus. MATERIALS AND METHODS: Cryopreservation of metaphase II human oocytes was performed using a fast-freeze, fast-thaw protocol. Upon thaw, oocytes were incubated for 3-4 h and then used for cytoplasmic donation (test oocytes). Oocytes thawed but not used for donation served as controls. Test and control oocytes were fixed using a microtubule stabilizing buffer. Tubulin was localized using antitubulin monoclonal antibody. Chromosomes were identified by counterstaining with DAPI. RESULTS: Forty-four oocytes had cytoplasm removed (test group) while 12 were not used for the procedure (controls). Twenty-three oocytes survived the donation procedure. Rates of normal spindle structure for the control and test groups were 21/23 (91.3%) and 12/12 (100%), respectively. CONCLUSION: The removal of cytoplasm from a metaphase II human donor oocyte does not appear to significantly increase the damage to chromosome alignment or to the spindle structure. PMID- 11432117 TI - Cryopreservation of mouse embryos at morula/compact stage. AB - PURPOSE: The objective of this article was to study mouse morula/compact stage embryo postthaw survival rates and postthaw developmental potential. METHODS: Mouse morula/compact stage embryos were classified into 3 substages. Their morphological behavior during the freezing and thawing process were examined. Postthaw survival rates and blastocyst formation rates were compared between the three substages and the pronucleate and the 2-cell stage embryos. Finally, Postthaw fully compacted and late compact embryos were transferred to foster mothers. RESULTS: Blastomeres of early compacting embryos separated from each other during the freezing process, whereas in fully compacted and late compact stages, the blastomeres remain compacted. Fully compacted and late compact stage embryos had higher Postthaw survival rates than other stages, though those were not statistically significant (P > 0.05). These two substages had significantly higher blastocyst formation rates than Postthaw pronucleate and early compacting embryos (P < 0.01). A total of 72 Postthaw fully compacted and late compact embryos were transferred and 20 live offspring were delivered. CONCLUSIONS: After passing early compacting stage, fully compacted and late compact stage mouse embryos have high Postthaw survival rates and significantly higher blastocyst formation rates. Successful cryopreservation of embryos at these two substages has a significant value in embryo selection. The results obtained in this study may provide useful information for human assisted reproductive technology. PMID- 11432118 TI - Differential diagnosis of dementia: clinical examination and laboratory assessment. AB - Recent breakthroughs in putative disease-modifying interventions for Alzheimer's disease (AD) underscore the urgency of making the earliest possible diagnosis. In the absence of a convenient and reliable laboratory test for AD, the clinical assessment is still the cornerstone of the diagnostic approach. This article provides a basis for conducting an assessment within the realities of a busy clinical practice for patients complaining of cognitive decline. The assessment will enable the clinician to diagnose the earliest manifestation of AD. PMID- 11432119 TI - Epidemiology and pathophysiology of Alzheimer's disease. AB - Researchers in Alzheimer's disease (AD) have made remarkable strides in the past 2 decades in characterizing the disorder, understanding its pathophysiology, and developing models for assessing treatment. With these successes and the promise of others to come, researchers and policy makers alike are keeping a nervous eye on the "demographic clock," which predicts that with the aging of the US population (and indeed the world), a massive increase in the number of AD cases and related dementias will emerge, flooding our medical care system and overwhelming our ability to care for these patients and their caregivers. This article explores some of the data underlying the explosion of AD and discusses the clinical disorder, its manifestations, and our advances in understanding the underlying pathobiology. Our growing knowledge of AD pathophysiology will lead us to the development of more effective medications to slow the progression or even prevent the emergence of this dreaded disorder. PMID- 11432120 TI - Treatment of Alzheimer's disease. AB - A growing consensus indicates that Alzheimer's disease (AD) results from an increase in the production or accumulation of beta-amyloid protein (A beta) leading to nerve cell death. Mechanisms by which A beta accumulation leads to neuronal death include oxidative damage and inflammation. This article discusses the management of AD patients with antioxidants, cholinesterase inhibitors, and psychotropic agents. Studies show that these agents can slow the progression of the disease, improve cognition, and reduce behavioral disturbances. A therapeutic alliance between physician and caregiver is an essential element in successfully managing the AD patient. The 3Rs--repeat, reassure, and redirect--can help caregivers reduce behavioral disturbances in patients with AD and limit the need for pharmacologic management. PMID- 11432121 TI - Dementia due to subcortical ischemic vascular disease. AB - Ischemic vascular disease (IVD) is the second most common cause of dementia in the Western world. This article focuses on dementia resulting from subcortical ischemic vascular disease (SIVD), a subtype of IVD, which in many cases may be prevented. Hypertension and diabetes are the leading causes of small-artery disease, subcortical brain ischemia, and stepwise or slowing progressive decline in cognitive function. The pattern of cognitive impairment in SIVD, as compared with Alzheimer's disease, is characterized by greater impairment of executive function but better preservation of recognition memory. Structural neuroimaging studies, such as computed tomography and especially magnetic resonance imaging, are more sensitive than the clinical examination and can enable detection of subcortical lacunes and deep white matter changes that are clinically silent. Often the brain can be protected against SIVD by early diagnosis and management of risk factors. Once end-organ damage has occurred, however, treatment outcome is less satisfactory. The most common risk factors for SIVD--hypertension and diabetes mellitus--are best detected and managed in primary care settings. PMID- 11432122 TI - Long-term care in dementia: patients and caregivers. AB - General principles of managing chronic, age-associated diseases apply as much to Alzheimer's disease (AD) and other late-life dementing disorders as they do to congestive heart failure or osteoarthritis. Beyond efforts to maintain residual tissue or organ function, important physician roles include promoting general well-being and helping patients and their caregivers adjust to disease-related limitations. Physicians provide essential information to patients and their families about the disease, its social and legal ramifications, and community resources to facilitate care. Therefore, physicians must be knowledgeable about broadly intersecting medical, legal, financial, and ethical issues surrounding the long-term management of AD and other dementias. The many challenges faced by patients with dementia and their caregivers over time underscore the need for an ongoing diagnostic and therapeutic alliance with primary care physicians. This article reviews salient aspects of long-term care for patients with AD and other dementias, highlighting the vital and varied roles of physicians in managing these chronic brain disorders. PMID- 11432123 TI - [Communication with, for and about the elderly]. PMID- 11432124 TI - Falls in the elderly. I. Identification of risk factors. AB - Falls severely threaten the health of elderly persons and pose high costs to the public health service. Unfortunately, falls are often regarded as unavoidable and untreatable features of aging. Therefore, many clinicians merely treat the physical injuries of a fall. However, falls and gait limitations are markers of underlying (sometimes otherwise subclinical) diseases that can be amenable to treatment. Moreover, falls and gait limitations herald the onset of repeated falls, physical decline, serious psychosocial consequences and a reduced survival. We review how clinically relevant risk factors can be traced by reviewing the medical notes, by careful history taking and by physical examination. The identified risk factors can serve as a template for the design of prevention strategies, which are discussed in the companion article. PMID- 11432125 TI - [Possible interaction between ethanol and drugs and their significance for drug therapy in the elderly]. AB - Even though alcohol dependence is not often found in the elderly, alcohol consumption and alcohol abuse are both common. As the elderly also often take medication on a regular basis, this group is at particularly high risk for problems resulting from the concurrent use of these substances. Physical changes as a result of the aging process (e.g. reduction of body water, decrease of hepatic blood flow) and alcohol related diseases can influence the pharmacokinetics and pharmacodynamics of both ethanol as well as other drugs. Alcohol dehydrogenase (ADH), acetaldehydede hydrogenase (ALDH) and cytochrome P450 2E1 are the enzymes responsible for the metabolism of ethanol. These enzymes are also the sites of direct pharmacological interaction between ethanol and other drugs, however, altered effects of medication can also be caused by ethanol adding to or reducing the drug's effect. Although some of these effects result from heavy use of alcohol, others can also occur with moderate use. Interactions have most frequently been described for analgetics, psychopharmacologically active drugs, antihistamines, anticoagulants antihypertensive drugs and antibiotics. PMID- 11432126 TI - Incidence and costs of osteoporosis-associated hip fractures in Austria. AB - This study is an investigation into the epidemiologic and socioeconomic impact of osteoporosis-associated hip fractures in Austria. We determined age- and gender specific incidence rates of osteoporosis-associated hip fractures for all patients treated in hospitals in 1995 and calculated mortality rates, hospitalization days and direct costs of hospitalization. The data were obtained from the hospital discharge statistics for all general hospitals and for all hospitals of the General Austrian Accident Insurance. To calculate the portion of hip fractures attributable to osteoporosis in a given age-group, a basic, non osteoporotic incidence of hip fractures was determined for ages 20-39, using gender-specific regression models. 11,379 patients with osteoporotic hip fractures underwent treatment in Austrian hospitals in 1995, accounting for 79 percent of all hip fracture patients treated. 82 percent of those were female, with the highest incidence among women aged 95 years and older with a rate of 3,000/100,000. For male patients the highest incidence was observed for the age group of 90-94 years with 1,743/100,000. International comparisons indicate these incidence rates to be similar to those reported for the Swiss population. In 1995, 778 patients or 6.8 percent of all patients with osteoporotic hip fractures died during hospitalization. Hospital care of patients with osteoporotic hip fractures required an overall 250,268 bed-days with an age-group-specific length of stay between 8.5-27 days for female and 16-23 days for male patients. The total cost of hospital treatment of osteoporotic hip fractures in Austria was ATS 1,043,379,000 (US$ 103,509,800), with average costs per patient of ATS 91,700 (US$ 9,097). Due to the aging of the population in the years to come, an increase of osteoporotic hip fractures among individuals aged 50 years and older must be expected. The economic importance of this development and its impact on the health care system must be considered as significant. PMID- 11432127 TI - [Coping with stress and blood viscosity in stroke prevention patients]. AB - INTRODUCTION: Hematocrit, fibrinogen and blood viscosity influence blood fluidity and are well known stroke risk factors. Studies have shown relationships between these factors and psychological stress. AIM: The aim of this study was to investigate how stroke risk patients with increased hematocrit, increased fibrinogen, or increased plasma viscosity differ from patients free of these risk factors in their ways of stress coping. METHOD: 6503 persons participated in the following stroke risk investigations: biographical and risk factor orientated anamnesis, neurological status investigation, laboratory investigation, sonographic investigation and psychological investigation. After assessment of several risk factors, differences in stress coping between risk factor and non risk factor groups were investigated by means of the t-test and the Wilcoxon test. RESULTS: Men with pathological hematocrit showed significantly higher scores in the coping strategy resignation and a tendency to less positive self instruction and response control attempts. Women with higher values of hematocrit demonstrated higher values in resignation and drug intake. Men with higher fibrinogen showed significantly higher scores in distraction, vicarious satisfaction, minimising by comparison and tendency to flee as well as a tendency towards drug intake. Women with increased fibrinogen showed no differences. Men with normal plasma viscosity had significantly higher values in tendency to flee and tendencially in desire for social support and lower values in minimising by comparison. Women with increased plasma viscosity demonstrated higher scores in resignation and aggression. CONCLUSIONS: The presence of elevated values in parameters of blood viscosity coincides with increased passive and defensive coping mechanisms, whereas non-risk factor persons show raised values in active coping styles. PMID- 11432128 TI - The management of elder mistreatment: the physician's role. AB - The physical abuse, psychological abuse, exploitation, and neglect of older adults constitute a serious and under-recognized public health problem throughout the world. Clinicians often misinterpret the health effects of elder mistreatment (EM) as caused by underlying disease or the aging process. Clues to mistreatment include the patient's appearance, recurrent urgent-care visits for the same diagnosis, missed appointments, suspicious physical findings, and implausible explanations for injuries. Avoiding confrontation and emphasizing treatment of abuse-related health conditions help the clinician maintain a therapeutic alliance with the victim and abuser. Victim safety should be the paramount concern. Victims with decisional capacity should be apprised of the chronic, progressive nature of EM. Clinical strategies to stop abuse include hospitalization and closer monitoring through office visits and home nursing. In most U.S. states, laws require that clinicians report at least physical abuse to the local adult protective services agency or to law enforcement. Mandated reporting, while offering potential social and legal remedies, raises ethical concerns regarding the physician-patient relationship. PMID- 11432129 TI - Completion angioscopy following carotid endarterectomy by the eversion technique or the standard longitudinal arteriotomy with patch closure. AB - INTRODUCTION: Technical defects are a potential cause of peri-operative strokes following carotid endarterectomy (CEA) by either the eversion technique or the standard method of longitudinal arteriotomy with or without patch closure. AIM: A non-randomised retrospective study was undertaken to assess the value of angioscopy for the identification of technical errors following CEA and to compare those detected following eversion and standard endarterectomy with patch closure. MATERIALS AND METHODS: Intra-operative completion angioscopy was performed following 110 CEAs (56 eversion, 54 standard). Angioscopy was omitted in 5 patients because of technical problems (2) or extreme shunt dependency (3). RESULTS: Significant technical defects were identified and corrected in 10 cases (9%) and were equally distributed between the eversion and standard procedures. There was no peri-operative stroke or death in this series. CONCLUSIONS: Significant residual defects may occur after CEA by either technique. Angioscopy allows their correction peri-operatively and has the potential to reduce peri operative stroke. PMID- 11432130 TI - The non-ischaemic blue finger. AB - Finger discoloration may result from recognized conditions affecting upper limb vasculature. We describe 11 patients who presented with acute pain, swelling and blue/purple discoloration in a finger. This benign condition mimicked digital ischaemia. There were 9 women and 2 men. The episode usually started with an ache/pain in the finger followed 2-3 h later by a blue/purple discoloration primarily on the volar aspect but always sparing the tip. This completely resolved after 4-7 days with no residual deficit. There was no history of trauma. Four patients had had previous episodes--2 had been started on warfarin. There was no family history and only one gave a history of spontaneous bruising of her legs. Examination of all patients--pulse rate, blood pressure, cardiac and subclavian artery auscultation and digital artery Doppler insonation--was normal. All patients had normal full blood counts, CRP, vasculitis screen and clotting (except those on warfarin). Six similar cases, all women, were reported in 1982. There was no common aetiological factor other than sex. Although of no prognostic significance, the condition is likely to concern patients and doctors in primary care. The discoloration is, however, clearly of a different distribution to that in an ischaemic finger where the tip of the digit is involved. PMID- 11432131 TI - Methicillin-resistant Staphylococcus aureus infection in vascular surgical patients. AB - BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) infection is emerging as a major problem in vascular surgical practice. The aim of this study was to review the management of patients with MRSA infection complicating vascular surgical operations. METHODS: Data were obtained from the vascular audit, case notes, intensive therapy unit (ITU) notes, high dependency unit (HDU) notes and microbiological records of patients who underwent either arterial reconstruction (n = 464) or limb amputation (n = 110) between April 1994 and October 1998. RESULTS: Forty-nine vascular surgical patients developed clinical MRSA infection (9%). Clinical MRSA infection in patients who had undergone aorto iliac reconstruction (n = 18) was associated with a 56% mortality (n = 10) and the most common infections were bacteraemia (55%) and pneumonia (50%). MRSA infection occurred in 17 patients who had undergone infra-inguinal bypass and was associated with a 29% mortality (n = 5). The most common site of MRSA infection was the groin wound (76%) leading to anastomotic dehiscence and death in one patient (11%) and necessitating wound debridement in 4 patients (22%). MRSA infection of the groin wound in the presence of a prosthetic graft (n = 3) led to anastomotic dehiscence in 2 patients, and graft excision in 2 patients. Similar complications were not observed in the presence of an underlying autogeneous long saphenous vein graft (n = 16). MRSA infection following major lower limb amputation (n = 14) was associated with death in 5 patients (36%). Wound infection in 10 amputees (71%) led to revision of the amputation to a higher level in 2 (14%) and wound debridement in 2 (14%). CONCLUSIONS: MRSA infection has a high mortality in vascular surgical patients in general, and following aorto-iliac reconstruction in particular. Autogeneous vein may confer some protection against local complications following groin wound infection. Strategies aimed at reducing the incidence of infection, including strict adherence to infection control procedures, may reduce the severity of this problem. PMID- 11432132 TI - Transmetatarsal amputation: an 8-year experience. AB - This study retrospectively reviews an 8-year consecutive series of transmetatarsal amputation (TMA) for forefoot ischaemia in diabetic and non diabetic patients. Forty-one patients had TMA. Peri-operative mortality was 17% (7/41). A healed stump was achieved in 19 patients (46%) and 18 of these patients were independently mobile, or mobile with sticks. Non-diabetic patients (8/12) healed significantly better than diabetics (11/29). Median time to healing was 7 months (range 3-20 months). All non-healed survivors had a higher amputation (14 below-knee, 1 Syme's). A healed TMA gives good mobility, but prediction of who will heal after operation is unreliable. Time to healing is often lengthy, and failed healing results in higher amputation. These issues need to be fully discussed with the patient who is considered for TMA. PMID- 11432133 TI - Should we continue oesophageal surgery in a district general hospital? A review of 200 consecutive cases. AB - A series of 200 oesophageal resections is presented. The results show that acceptable standards are obtainable in a district general hospital (DGH) thus favouring the suggestion that such services could be organised by 'networking' rather than centralisation. This avoids the disadvantages to many DGH patients which follow centralisation. PMID- 11432134 TI - Detection of Crohn's strictures--the use of an 'Endo-marble'. PMID- 11432135 TI - Colorectal patients and cardiac arrhythmias detected on the surgical high dependency unit. AB - INTRODUCTION: Surgical high dependency unit (SHDU) care is becoming an integral feature of colorectal surgical practice. Routine ECG monitoring is a feature of surgical care in this setting. The aim of this study was to determine the incidence and outcome of cardiac arrhythmias detected in an SHDU population of colorectal patients. PATIENTS AND METHODS: 226 patients over a 12 month period were admitted to a 6-bedded SHDU under the care of 3 colorectal surgeons. A total of 29 patients (13%) had significant arrhythmias on ECG monitoring (median age 74 years, range 35-88 years). Pre-existing ischaemic heart disease was present in 9 patients--colorectal cancer and inflammatory bowel disease accounted for the underlying problem in the majority of these patients. RESULTS: Equal numbers of supraventricular and ventricular arrhythmias were detected--atrial fibrillation being the most commonly detected abnormality. Therapeutic intervention (electrolyte correction and anti-arrhythmic agents) was required in 23 patients. One patient required DC shock for ventricular fibrillation. Seven patients were transferred to the heart care unit or intensive care unit to manage their cardiac problems. Two patients died as a result of their cardiac problem, 27 were discharged home alive--3 on long-term anti-arrhythmic therapy. CONCLUSIONS: The postoperative environment of colorectal patients has been radically altered by the introduction of the SHDU. If colorectal surgeons are to remain central to the postoperative care of their patients, all surgical staff will require training in the recognition and protocol prevention and management of cardiac arrhythmias. Certification of colorectal surgeons in advanced life support is more relevant to colorectal surgery than certification in trauma care. PMID- 11432136 TI - Post vasectomy analysis: call for a uniform evidence-based protocol. AB - BACKGROUND: Vasectomy is a safe and reliable means of contraception that is used by 42 million couples world-wide. There are no standardised guidelines in the follow-up of these patients to assess the efficacy of the procedure, however. Through this paper we aim to recommend a guideline that is both clinically effective and cost effective, and would avoid medicolegal debate. PATIENTS AND METHODS: A questionnaire was sent to consultant general surgeons and urologists working both within and outside the North-West region, to assess the range of protocols used in the follow-up of patients undergoing vasectomy. RESULTS: Of respondents, 75% sent the vas deferens routinely for histology: 90.4% requested 2 sperm samples routinely whilst 4.8% requested 1 sample and 4.8% requested 3 samples. 95.2% requested further sperm samples if non-motile sperm were present. 98.8% performed their first sperm sample by 12 weeks. CONCLUSIONS: There is a wide range of protocols used in the follow-up of patients post-vasectomy. Most do not appear to be evidence-based. We recommend a guideline which re-emphasises the importance of pre-operative counselling, only 1 routine sperm sample taken 16 weeks post-vasectomy, and further samples taken only if motile sperm is seen. Non motile sperm is not an indication for further sperm samples. We believe this guideline would be both clinically effective and cost effective. PMID- 11432137 TI - Penile cancer: a case for guidelines. AB - INTRODUCTION: Aspects of the management of penile cancer remain controversial. In the management of early T1 N0 disease, treatments are divided between amputation and a variety of penis conserving techniques (PCT); local excision, laser techniques, chemotherapy and radiotherapy. We report on a retrospective series of patients with penile cancer. PATIENTS AND METHODS: Thirty-seven patients were diagnosed between 1987-1996. All patients records were retrieved. Data recorded included TNM stage, histological grade and treatment. The end-points were death, nodal progression and local recurrence. RESULTS: Median survivor follow-up of 42 months was obtained. Twenty-six patients (70%) presented with T1 disease, 7 (19%) T2 and 4 (11%) T3 or T4. Inguinal nodal disease was seen in 11 (30%). The mean age was 63 years. Overall, 13 penile amputations were performed, 13 underwent radiotherapy, 6 were locally excised in combination with radiotherapy and 3 underwent local excision alone. Two patients were unsuitable for treatment. Of the total (37 patients) 15 have died; 12 from penile cancer. Ten have suffered disease progression and 12 remain alive with no evidence of disease. Twenty-three patients presented with early T1 NO disease. They were treated with radiotherapy (12), local excision (2), combined radiotherapy and excision (2) and partial amputation (4). Outcome was not significantly related to treatment modality. Spread to the inguinal nodes or local recurrence has occurred in 10, of whom 2 have died. Only 13 (57%) appear disease-free. CONCLUSIONS: The characteristics of the patients and the disease in this series are similar to published series in Europe and North America. There is significant variability in the modalities of treatment used within this series. Local recurrence and disease progression occurs in 43% of T1 N0 lesions. There would seem to be some room for improvement. International data are retrospective and inconclusive with regard to best practice. There is an urgent requirement for randomised controlled trials to improve the outcome of these patients. PMID- 11432138 TI - The orthopaedic presentation of acute leukaemia in childhood. AB - In children, acute leukaemia can mimic several orthopaedic pathologies and this variable presentation creates difficulties in achieving the correct diagnosis. Four consecutive cases are reported, all of which initially presented via the orthopaedic department, representing 17% of all new cases of leukaemia presenting to our hospital. This series is presented to highlight the need to place acute leukaemia on the list of differential diagnoses of acute joint pathology in children: the associated literature is reviewed. PMID- 11432139 TI - Primary total hip replacement: variations in patient management in Oxford & Anglia, Trent, Yorkshire & Northern 'regions'. AB - OBJECTIVES: To examine national practice, and variations in practice, concerning total hip replacement; in particular the choice of prosthesis and the involvement of consultants in NHS operations. DESIGN: Pre-operative survey of patients undergoing total hip replacement. SETTING: Five English regions serving combined population of 16.8 million people. SUBJECTS: 13,343 total hip replacement operations in one year commencing September 1996, either in NHS or private sector. MAIN OUTCOME MEASURES: Prosthesis used for surgery, status of surgeons involved in operation, use of laminar flow operating theatre. RESULTS: Prostheses without well documented 5-year survival were used in 5504 (58%) of 9417 operations for which information was available. The consultant was the operator in 4810 (64%) of 7499 NHS operations. In 1352 trainee-led operations, the consultant was present for only 637 (47%); this figure was 54% for trainees in years 1-4 of their training. Substantial variation between NHS consultant firms occurred both for use of prostheses with well documented survival data, and supervision of trainees by the consultant. CONCLUSIONS: This large study is the first attempt to describe national practice for primary total hip replacement. Substantial variation among consultant firms was observed for all indices of practice reported. PMID- 11432140 TI - Hunterian lecture. Study of dermal grafts and cultured autologous keratinocytes in an experimental model. PMID- 11432141 TI - Clinical features of soft tissue sarcomas. AB - The presenting features of 526 patients referred to an open access 'lumps and bumps' clinic were reviewed to try and identify whether the four cardinal features of soft tissue malignancy were in fact predictive of this. The features investigated were: size bigger than 5 cm, pain, increase in size, depth beneath the deep fascia. All of these factors were found to be associated more frequently with malignancy than a benign state. Using the summed weights of evidence method we have constructed a graph which will allow prediction of whether a lump is likely to be malignant or not. PMID- 11432142 TI - Lateral repair of parastomal hernia. AB - INTRODUCTION: Parastomal hernia is a common complication of stoma construction. Although the majority of patients are asymptomatic, about 10% require surgical correction. AIMS: We describe a new surgical approach for the repair of parastomal hernias, which avoids both the need for laparotomy and stoma mobilization. PATIENTS AND METHODS: Nine patients (4 female) with parastomal hernia underwent surgical repair. Median age was 55 years (range 38-73 years). There were 8 para-ileostomy herniae and one paracolostomy hernia. A lateral incision was made approximately 10 cm from the stoma, and carried down to the rectus sheath. The dissection was carried medially towards the stoma, and around the defect in the abdominal musculature. The hernia sac was excised when possible and the fascial defect closed with non-absorbable, monofilament suture. A polyprolene mesh was placed round the stoma by making a slit in the mesh. The skin was closed with subcuticular monofilament absorbable suture. RESULTS: All patients returned to normal diet on the first postoperative day, and were discharged from hospital within 72 h. There were no wound infections, and no recurrences after a median follow up of 6 months (range 3-12 months). DISCUSSION: The technique we describe is simple and avoids the need of laparotomy. The mucocutaneous junction of the stoma is not disturbed, reducing the risk of contamination of the mesh, stenosis or retraction of the stoma. Grooving of the stoma and difficulty in fitting appliances is avoided because the wound is not placed near the mucocutaneous junction. This approach may be superior to other mesh repairs for parastomal hernia. PMID- 11432143 TI - Guidelines for inguinal hernia repair in everyday practice. AB - BACKGROUND: The Royal College of Surgeons of England published clinical guidelines for the management of groin hernia in adults in July 1993. We compared our indications, techniques, complications and outcome with these guidelines. PATIENTS AND METHODS: A consecutive series of 440 patients who underwent a groin hernia repair from the 1 July 1994 to 30 July 1995 were studied retrospectively. Special consideration was given to the advantages and acceptance of day-case surgery. Confidential questionnaires were sent to all patients 6-12 months following surgery. RESULTS: The majority of elective primary inguinal hernias (83%) were repaired by the open tension-free Lichtenstein mesh technique. Our selection criteria for day-case surgery included ASA I, age (< 65 years) and social situation; 56% underwent an operation on a day-case basis. Including in patients discharged within 24 h following operation, this proportion of 'day cases' increased to 72.5%. Less complications occurred in the day-case group (P = 0.018). However, this difference may be caused by incomplete reporting of complications in the day-case group. There were no significant differences in patients' satisfaction, postoperative attendance for medical advice or time back to work between the day-case and in-patient group. CONCLUSIONS: The guidelines need to be reviewed. The Lichtenstein repair offers an excellent and simple technique for hernia repair as a day-case procedure. Our results suggest that the number of hernia repairs performed as a day-case could substantially be increased to more than the recommended 30%. Time off work is mainly influenced by the advice given by GPs and surgeons. Reducing time off work by giving more appropriate recommendations and increasing the number of day-case surgery cases could significantly reduce the costs of health-care. PMID- 11432144 TI - The realities of postoperative disability and the carer's burden. AB - Outcome after high-risk, complex neurosurgery for progressive skull base pathology, and its effect on carers, has been examined. Two different outcome measures were used. The Glasgow Outcome Score (GOS) assesses overall social capability and dependence of the patient, while the 36 item short-form health survey (SF-36), a generic quality of life measure, can be compared directly with the general population. Overall outcome using the GOS indicated a favourable outcome for 13 of the 17 patients studied. The SF-36 demonstrated that more than half the patients were functioning at a level below the accepted norm. The reasons for this discrepancy and the validity of outcome scales have been analysed. In addition, the effect upon carers, its relevance to assessment of outcome, and the need to involve potential carers in the process of informed consent was stated. Our conclusions are applicable throughout the surgical specialities, and especially to high-risk complex surgery. PMID- 11432145 TI - Insertion of self-expanding metal stents for malignant dysphagia: assessment of a simple endoscopic method. PMID- 11432146 TI - Audit of topical glyceryl trinitrate for treatment of fissure-in-ano. PMID- 11432147 TI - Standardisation of the selection process for applicants for SHO posts for basic surgical training rotations. PMID- 11432148 TI - A prospective randomised trial of tourniquet in varicose vein surgery. PMID- 11432149 TI - Stats & facts. The many sentinels of health plan quality. PMID- 11432150 TI - Digital tips for the medical E-customer. PMID- 11432151 TI - Fiscal options for America's best hospitals. AB - A review of America's "best" teaching hospitals shows a huge disparity in their fiscal positions. Among the 15 hospitals studied, roughly half experience some fiscal distress. However, a somewhat similar fiscal analysis of the nation's largest investor-owned hospital chains, HMOs, and physician practice management corporations shows an even more serious weakness in operating margins and debt equity ratios. Aside from raising possible ethical, quality, and cost issues, this financial analysis suggests that conversion from nonprofit to for-profit ownership of America's top teaching hospitals might not guarantee an improvement in their long-term fiscal outlook. PMID- 11432152 TI - The Academy of Managed Care Pharmacy's format for formulary submission. PMID- 11432153 TI - Showing outcomes and proving value brings success. AB - Today's pharmacoeconomics have shown how critical it is to find value in formulary drug products. For Regence BlueShield, Seattle, this means outcomes; making sure a drug truly works better than well-established alternatives, not just placebos. In 1998, Regence adopted formulary submission guidelines that have proven a solid success, and have been adopted by the Academy of Managed Care Pharmacy, Alexandria, Virginia, for evaluation nationwide. PMID- 11432154 TI - Disease management, outcomes, and medical errors. PMID- 11432155 TI - Providing high-quality customer service during a time of crisis. AB - The tumultuous fall into receivership of Harvard Pilgrim Health Care, and its subsequent financial rise, has been well documented. One of the principal concerns in situations of financial upheaval is how to ensure that staff and customers maintain a positive attitude toward the troubled health plan. An insider at Harvard Pilgrim provides a glimpse at the steps that were taken to ensure high-quality customer service and to retain valuable staff. PMID- 11432156 TI - Putting to use a clinical guideline for treating tobacco dependency. AB - The issuance last year of the U.S. Public Health Services' clinical guideline for treating tobacco use and dependency offered managed care a tool that could help them achieve the Healthy People 2010 goals of cutting adult smoking rates in half. The authors describe how the guidelines approach this difficult public health issue. PMID- 11432157 TI - Emergency Medical Treatment and Labor Act compliance for hospital clinics. AB - The Emergency Medical Treatment and Labor Act imposes a number of requirements on hospital outpatient treatment departments, including the assurance of patient access to hospital care for providers participating in the Medicare program. The author describes the key provisions of the legislation, including hospital staffing and policy requirements. PMID- 11432158 TI - The automated electronic defibrillator--should every office be required to have one? PMID- 11432159 TI - The hypoalgesic effect of 3-D video glasses on cold pressor pain: reproducibility and importance of information. AB - The first aim of the study was to evaluate whether it was possible to manipulate the distraction effect induced by 3-D video glasses on the perceived pain and unpleasantness of the subjects by giving them different information about the expected effect. Second, the study aimed to determine the reproducibility of the effect. Forty-five students enrolled, 39 students participated in the study (24 women and 15 men, median age 23 years, range 19-28 years) because 6 did not show up for the first trial, and 37 completed the study because 2 subjects did not show up for the second trial. The subjects were randomized into 3 groups, balanced with respect to age and sex, that received different information about the effect of 3-D video on pain and unpleasantness: the first group received positive information, the second group received neutral information, and the third group received negative information. Once assigned to a group, there were no crossovers between the groups. A cold pressor stimulus was used to induce experimental pain, and the volunteers rated the intensity of pain and unpleasantness on 100-mm visual analogue scales. A new generation of video glasses were used in the study. Each volunteer was exposed to the cold pressor test in 2 randomized trials (video and control) after the information was given, and the 2 trials were repeated in a second session after 4 weeks. There was no significant difference in the effect of video glasses on perceived pain (P = .74) nor on the perceived unpleasantness (P = .84) among the 3 information groups. The data were therefore pooled. The results of the pooled data showed a significant effect of 3-D video on perceived pain (P = .03) but not on unpleasantness (P = .18). After 4 weeks, the study was repeated, and there were no significant changes in the effect of video glasses. The median visual analogue scale scores were reduced in both the video and the control trials compared with the first session. PMID- 11432160 TI - The use of bispectral analysis to monitor outpatient sedation. AB - The bispectral (BIS) index has been used to interpret partial EEG recordings to predict the level of sedation and loss of consciousness in patients undergoing general anesthesia. The author has evaluated BIS technology in determining the level of sedation in patients undergoing outpatient deep sedation. These experiences are outlined in this review article. Initially, the correlation of the BIS index with traditional subjective patient evaluation using the Observer's Assessment of Alertness and Sedation (OAA/S) scale was performed in 25 subjects. In a second study, the recovery profile of 39 patients where the BIS was used to monitor sedation was compared with a control group where the monitor was not used. A strong positive relationship between the BIS and OAA/S readings was found in the initial subjects. From the recovery study, it appears that use of the BIS monitor may help titrate the level of sedation so that less drugs are used to maintain the desired level of sedation. A trend to earlier return of motor function in BIS-monitored patients was also demonstrated. BIS technology offers an objective, ordinal means of assessing the depth of sedation. This can be invaluable in comparing studies of techniques. The BIS index provides additional information to standard monitoring techniques that helps guide the administration of sedative-hypnotic agents. The trend to earlier return of motor function in BIS monitored patients warrants further investigation. PMID- 11432161 TI - Additional clinical observations utilizing bispectral analysis. AB - Additional observations were made in the use of the bispectral (BIS) index with the use of ketamine and in performing general anesthesia without the use of local anesthesia in nonintubated patients. Twenty-five subjects undergoing extraction procedures in an outpatient setting were analyzed using bispectral analysis with ketamine and midazolam. Despite repeated injections of midazolam during the procedure, only transient decreases of the BIS occurred to the 80s, with a low value of 77 in all but 1 patient where ketamine was used. In comparison, values in the 50-70 range are typically seen immediately after the administration of sedative doses of midazolam, propofol, or methohexital. In the second study, once propofol anesthesia was initiated, BIS readings in the 30s were commonly seen in patients during their procedure. The lowest BIS level observed was 18. Bispectral analysis was useful to trend the present anesthetic state and adjust the dose of propofol accordingly. In no case was laryngospasm or total airway obstruction observed. In 1 case, partial airway obstruction secondary to retro-positioning of the tongue occurred with a subsequent decrease in oxygen saturation to 89%. This was rectified by repositioning the patient to alleviate the obstruction. Consistent with previous studies utilizing ketamine, BIS values are consistently higher when compared with other hypnotic agents. With the subsequent injection of midazolam, the BIS level did not decrease to anticipated levels. In the final study reviewed, when local anesthesia was not used during general anesthesia, bispectral analysis was a useful adjunct in helping maintain a steady state of general anesthesia in nonintubated patients undergoing third molar extractions. Bispectral analysis offers additional information on the depth of the hypnotic state and is useful in helping control the depth of anesthesia. A limitation of the index includes the inability to titrate the level of sedation induced by hypnotic agents such as midazolam when ketamine is concomitantly administered. PMID- 11432162 TI - [Esophageal anastomosis--based on the experience with 1460 operations]. AB - The authors reviewed the clinical records of 1460 patients with esophageal anastomoses, the operations performed in their institute between 1973 and 2000. Surgical outcome was assessed with incidence of anastomotic leaks and overall mortality. Hand-sewn anastomoses were performed by inserting single layer interrupted monofil steel wire (507), PDS (232) or Vicryl (65) sutures and circular stapler (EEA, Ethicon) was used in 656 patients. 453 (65 cervical, 293 thoracic, and 95 abdominal) anastomoses performed between 1995 and 2000 were analyzed separately. During this 6-year period, the majority of operations (88%) were performed because of esophageal malignancy (squamous cell carcinoma of the esophagus or adenocarcinoma of the cardia). Surgery was performed for benign disease (e.g. stricture, perforation, and stage IV achalasia) in 12%. The incidence of anastomotic leak was the highest (20%) after cervical anastomosis, compared to a 4.4% or 1% associated with thoracic or abdominal anastomoses. These differences are statistically significant (p < 0.001). The rate of anastomotic leakage was strongly connected to surgical technique: it was 15% in hand-sewn anastomoses and only 4.4% with stapled anastomosis. During the 6-year period overall mortality was 6.6%t. PMID- 11432163 TI - [Perioperative treatment after esophagogastric surgery]. AB - Extended gastric and esophageal resection is still associated with high postoperative morbidity and mortality. We performed a retrospective analysis of the perioperative management of 72 patients who had undergone such operations during a one-year period. Patient and management variables were analyzed by multivariate statistical methods to identify pre-, intra-, and postoperative factors which influence the results. The investigation of preoperative data revealed an increase of esophageal cancer among younger (< 55 years) patients (12 patients underwent gastrectomy and 22 esophageal resection). The intensity of smoking was significantly higher compared to the elderly (11.27 vs. 7.4 cigarettes/day; p < 0.05); and the same applies to alcohol consumption. In older patients (> 55 years of age), the duration of postoperative artificial ventilation was significantly longer (10.1 vs. 4 hours, p < 0.05) and the prevalence of septic complication was higher, than in younger patients. All three postoperative deaths recorded in this series occurred in the group of elderly patients. A preoperative weight loss exceeding 10 per cent of body weight was associated with significantly longer postoperative stay (21.6 vs. 17.4 days; p < 0.001), as well as with need for longer parenteral feeding (13.05 vs. 10.06 days; p < 0.005). Operations longer than 6 hours were associated with significantly longer postoperative ventilation period (14.44 vs. 5.31 hours; p < 0.02), need for longer stay intensive care unit (10.56 vs. 6.55 days; p < 0.001) and longer postoperative stay (21.56 vs. 17.64 days; p < 0.05). The prevalence of pulmonary complications was connected to the duration of the operation (10/16 vs. 3/55). We also describe and analyse two contemporary methods designed for monitoring circulatory parameters (PICCO) and tissue oxygenation (gastrotonometry). The analysis of postoperative data demonstrates that postoperative pain control with continuous epidural analgesia is superior to methods as it shortens the length of stay on the intensive care unit (7.15 vs. 10.67 days; p < 0.05) and postoperative hospitalisation (18.06 vs. 23.50 days; p < 0.05). Nutritional support is essential after esophageal anastomosis till oral feeding can start. Enteral nutrition was given through a jejunal tube that had been inserted intraoperatively. Calorie intake was built up step by step to a maintenance level of 31.2 kcal/day, which was administered until oral feeding could be started (mean duration 10.94 days; maximum duration: 42 days). We conclude that careful selection of patients, appropriate intra- and postoperative management, with adequate postoperative pain control can reduce postoperative morbidity and length of inpatient stay. PMID- 11432164 TI - [Siewert-Stein classification of adenocarcinoma of the esophagogastric junction]. AB - Nowadays the terminology used for the definition of adenocarcinomas at the oesophagogastric junction is "cardiac carcinoma", which can be easily misunderstood. This definition of adenocarcinomas of the oesophagogastric junction does not allow correct comparison of diagnosis (endoscopic, radiological and pathologic), epidemiology and surgical therapy in national and international aspects, because different tumours can develope in the same area, and all called cardia tumors. Siewert and Stein recommended a classification to solve this problem. The classification of the tumours is morphological/topographical. Type I is adenocarcinoma of the distal part of the oesophagus. Type II is adenocarcinoma of the real cardia and type III is subcardial gastric adenocarcinoma. At classification, we always consider results of endoscopy (ortograde and retroflexed view of the oesophago-gastric junction), the x-rays of the oesophagus and stomach, findings at the operation and pathohistologic results. Between 1/1/1974 and 31/12/2000, a total number of 50,878 upper panendoscopic examinations were performed at the Endoscopic Laboratory of the Surgical Department. Adenocarcinoma of the cardia was diagnosed in 488 patients. According to the Siewert-Stein classification, type I tumour was found in 123 (25.2%), type II in 240 (49.18%), and type III was present in 125 (25.61%) patients. The importance of this classification is it enables unified pre-operative assessment and it can also help to decide the type of the surgical intervention. In our patients with type I cancer--depending of the size of the tumour--distal 2/3 oesophagectomy with the resection of the proximal lesser curve of the stomach or total gastrectomy were performed. In the first group oesophago-jejuno gastrostomy, in case of total gastrectomy Roux-en-Y loop anastomosis was created. In patients with types II and III cancers total gastrectomy was performed. In every patient lymphadenectomy was performed. We suggest the use of this new classification in clinical, gastroenterology--with special regard to the endoscopy--and pathology. PMID- 11432165 TI - [Perspectives of laparoscopy in cardia and gastric surgery]. AB - The authors have made 171 laparoscopic operations in the gastro-esophageal region during the past three and a half years. They have used the achievement of the minimal invasive surgery mainly in the treatment of functional diseases of the gastro-esophageal junction. Besides the hiatal reconstruction and antireflux procedures (142), endoscopic assisted cysto-gastrostomy (7), endoscopic assisted double-lifting gastric wall resection (5), cardiomyotomy (4), Graham suture of perforated duodenal ulcer (4) and enucleation of gastric wall leiomyoma (2) has been performed. There was no postoperative death, the rate of conversion was altogether 4.7% (8/171) while there was no need for reoperation in the early postoperative period. PMID- 11432166 TI - [Endoscopic diagnosis of gastrointestinal bleeding of unknown origin]. AB - In recent years push enteroscopy has become the most important method in the examination of patients with obscure gastrointestinal bleeding. We summarise our experiences with 148 enteroscopies performed on 140 patients with bleeding of unknown origin. The source of bleeding could be identified in 81 patients (57.86%). The most common lesions were small bowel tumors (13.57%) and vascular malformations (12.86%). Several patients (22.86%) referred for enteroscopy had lesions in the esophagus, stomach and proximal duodenum that were missed at upper endoscopy. The authors conclude that push enteroscopy has a major role in the evaluation of patients with obscure gastrointestinal bleeding. Enteroscopy should be the first diagnostic step after negative esophago-gastroduodenoscopy and negative colonoscopy. Enteroscopy seems to be superior to other diagnostic methods. It is well tolerable and is not time consuming. PMID- 11432167 TI - [Results of surgical treatment of esophageal cancer]. AB - At the Department of Surgery of the Szent Imre Hospital 53 esophageal operations were performed between August 1, 1995 and August 1, 2000 because of malignant disease. We retrospectively analyze the operations of 37 male and 16 female patients, and compare the results with published data. We describe the preparation and medical examination of the patients and the problems of intraoperative or potential complications and their prevention. We emphasise the importance of postoperative treatment and monitoring, as well as difficulties of the surgery and physiotherapy after operation. We think, that with adequate perioperative background and surgical treatment these operations can be performed safely in our department. These are basic procedures in the treatment of a malignant disease of the digestive system with serious economico-medical problems. PMID- 11432168 TI - [Laparoscopic adrenalectomy with transperitoneal approach]. AB - The feasibility, safety, and results of 52 laparoscopic transperitoneal adrenalectomies were evaluated. METHODS: A total of 52 patients were included in the study based on thorough endocrinological and imaging assessment. 15 patients with Conn syndrome, 3 with Cushing syndrome, 15 with nonfunctioning adenoma, 14 with pheochromocytoma, 2 with adrenocortical cyst, 2 with adrenocortical lipoma and 1 with metastasis were considered eligible for adrenalectomy. Lesion size ranged from 1 to 12 cm (mean 4.53 cm). Concurrent surgical procedures were performed in 6 patients (11%). RESULTS: There was one conversion (during a left adrenalectomy), because of our learning curve. After we changed the technique, there was no more conversion. There were two (3.8%) postoperative complications: postoperative pancreatitis, one of the patients required re-operation (lavage and drainage). There was one wound infection. We had no postoperative mortality. Mean postoperative hospital stay was 6 days (range, 2-27 days). CONCLUSION: Patients with secreting and non-secreting adrenal lesions can be treated safely and effectively by laparoscopic adrenalectomy. PMID- 11432169 TI - [Possibilities in the preservation and restoration of anal continence]. AB - The preservation of anal continence and the improvement of the patients' quality of life in general are primary objectives of colorectal surgery. Earlier the loss of the entire rectum, colon required a definitive stoma. This review describes surgical procedures designed to preserve anal continence. This paper also describes operative techniques designed to improve impaired sphincter function. Total extirpation of the mesorectum reduces local recurrence of rectal tumours. At the same time, this operation requires formation of the anastomosis low, at the level of the levator muscle. Low colorectal or coloanal anastomoses are associated with higher incidence of suture leakage and poor functional outcome. The distance between anastomosis and anal verge was less than 7 cm in 249 sphincter-sparing rectal resections performed during the examined 6-year period in our institute. Different techniques to perform anastomoses were applied, the prevalence of suture leakage and the functional results are analysed. Restorative proctocolectomy has dramatically improved the treatment of familial polyposis and ulcerative colitis with rectal involvement. Although proctocolectomy is necessary to cure the disease, acceptable faecal continence can be achieved by creating ileoanal anastomosis with ileal reservoir. We discuss our results after 43 operations. Weakness of the sphincter apparatus is the most common cause of continence problems. Occasionally, the sphincter is no longer suitable for reconstruction because of extensive damage or denervation. In such cases, the levator muscles or--if neither these are of acceptable quality--the gluteus maximus muscle can be used to repair the external sphincter. Anterior levator plasty involves tightening the levator plate by suturing its arches together between the rectum and the vagina. This procedure enhances the resistance of the sphincter barrier primarily by increasing functional sphincter length. The functional outcome of this procedure was acceptable in two-thirds of the 52 operations. Post anal repair was performed only in 3 patients. This method comprises reinforcing the levator plate through an access between the external and the internal sphincters. When the levator plate is unsuitable, bilateral gluteus plasty can be performed to increase the strength of sphincter muscles. As the gluteus is a striated muscle it can improve only the of the external sphincter function. Therefore this procedure can restore acceptable continence to hard stool only. This is demonstrated by our clinical experience obtained in 10 patients. PMID- 11432170 TI - [Laparoscopic colon surgery]. AB - We review our experience in laparoscopic colorectal surgery, with indications, technical aspects and results. Between 1992 and 31/12/2000, we performed 113 laparoscopic or laparoscopically assisted colorectal operations. Of 79 malignant cases, 37 operations were oncologically radical and therapeutic, 42 were palliative. During the immediate postoperative period two deaths occurred (2.8%), the causes of death were not related to surgery. Port site metastasis developed in one patient (1.4%). Postoperative complications developed in 18 patients (14.5%). Only one patient required conversion to laparotomy. We emphasize the importance of hand assisted laparoscopic surgery (HALS) in laparoscopic colorectal surgery, because it can increase the number of laparoscopic colorectal operations. Based on our results and experience, we recommend the routine use of laparoscopic technique in colorectal surgery. PMID- 11432171 TI - [How do intraoperative complications effect adhesion formation after laparoscopic cholecystectomy: a comparative animal study]. AB - BACKGROUND: The aim of this study was to evaluate the extent of postoperative formation of adhesions following laparoscopic and open cholecystectomy. MATERIAL AND METHODS: 60 experimental laparoscopic cholecystectomies (LC) were performed in dogs by qualified surgeons to learn laparoscopic technique. To assess the relationship between complications occurred during the operation (bleeding, laceration of the liver bed or gallbladder perforation) and the formation of adhesions surviving animals were divided into 4 groups according to the type of complication. We assessed the results during second-look laparoscopy 4 weeks following LC using the adhesion index (AI: 0-4 score). Animals were then sacrificed to measure the extent of adhesions. As a control group open cholecystectomy was performed in 15 dogs without intraoperative complications. Mann-Whitney Rank Sum test and Dunn's Method were used for statistical analysis. RESULTS: No adhesions were observed in the laparoscopic group without intraoperative complications. In all dogs with bleeding or laceration of the liver bed maintained by electrocoagulation, adhesions developed. Formation of adhesion in these groups was significantly higher than in "ideal LC" or in case of gallbladder perforation (P < 0.01). All animals in the control group developed significantly more adhesions compared to the experimental group (p < 0.05). CONCLUSION: LC produces less adhesion compared to conventional open technique. Complications such as bleeding or laceration of the liver bed during LC can increase the formation of adhesions. No formation of adhesions can be related to gallbladder perforation during LC. PMID- 11432172 TI - [Effect of pentoxifylline on the healing of experimental anastomosis of the left colon in rats]. AB - The healing of colonic anastomoses is determined by several factors such as microcirculation, the strength of the inflammatory response, and the time required for regeneration. We investigated the effects of pentoxifylline--a drug which improves microcirculation and modulates leukocyte functions--on the healing of experimental anastomosis on the left colon of rats. As a result of drug treatment (0.25 mg/100 g, i.p.) in Group I anastomosis bursting pressure (ABP) was by 56 +/- 17% higher at day 2 than in controls with no pentoxifylline treatment. On the 5th postoperative day in Group I, ABP reached 80 +/- 8% the value for the intact colon (218 +/- 21 mmHg), whereas respective value in the control (untreated) group was only 47 +/- 7%. In Group II (pentoxifylline: 2 mg/100 g, i.p.) ABP was by 55 +/- 10% and by 73 +/- 8% higher than control values at postoperative days 1 and 2, respectively. At day 2, in Group I colonic blood flow measured at the anastomosis line by 86Rb uptake technique was significantly higher than in the untreated controls (0.18 +/- 0.01 ml/min vs. 0.14 +/- 0.01 ml/min, (p < 0.02). Blood flow measured in colon tissue above and below the anastomosis changed differently. Pentoxifylline treatment also suppressed the peritoneal inflammatory response assessed with peritoneal reaction index (2.0 +/- 0.3 vs. 1.1 +/- 0.2, p < 0.01). The results of the present study show that pentoxifylline treatment shortens the time needed for the healing of colonic anastomosis. These observations suggest that pentoxifylline medication can prevent failure of colonic anastomoses. PMID- 11432173 TI - [Rare late complication after subtotal esophagectomy]. AB - We report a case of a peptic ulcer developed in the stomach tube used for the replacement of the esophagus. The patient was a 60 years old female who had undergone subtotal esophagectomy for mid esophageal malignancy, with intrapleural stomach replacement. Urgent endoscopy revealed an excavated, bleeding ulcer in the thoracic part of the stomach. After unsuccessful medical treatment urgent operation was performed via right thoracotomy. Opening the stomach an ulcer was found on the posterior wall of the stomach, it was penetrating to the right atrium of the heart. The bleeding was controlled by suturing the atrium wall. The patient treated with i.v. Omeprazol in the postoperative period. On the 21st postoperative day a rebleeding occurred causing shock. After reoperation the patient died. This complication is very rare. We emphasise the importance of postoperative pH measurement investigations showing the presence of duodenogastric reflux disease. PMID- 11432174 TI - [Isolated gastroduodenal Crohn's disease: a case report]. AB - Although it can involve any segment of the gastrointestinal tract, Crohn's disease confined to the gastroduodenum is extremely rare. We report the story of a 20-years old male patient admitted for pyloric obstruction that developed despite medical treatment. Clinical manifestations necessitated operative treatment; Polya-gastrectomy was performed. Histology identified Crohn's disease and inflammatory changes as the cause of pyloric obstruction. Postoperative recovery was uneventful. The lesson of this case is, that if duodenal Crohn's disease would have been recognized preoperatively, gastrectomy could have been avoided. We review the literature and discuss treatment options for gastroduodenal Crohn's disease. PMID- 11432175 TI - The steel-needle butterfly--an outdated intravenous lifeline technology in modern anesthesiology. PMID- 11432176 TI - Preemptive effects of a combination of preoperative diclofenac, butorphanol, and lidocaine on postoperative pain management following orthognathic surgery. AB - The aim of the study was to investigate whether preemptive multimodal analgesia (diclofenac, butorphanol, and lidocaine) was obtained during sagittal split ramus osteotomy (SSRO). Following institutional approval and informed consent, 82 healthy patients (ASA-I) undergoing SSRO were randomly assigned to 1 of 2 groups, the preemptive multimodal analgesia group (group P, n = 41) and the control group (group C, n = 41). This study was conducted in a double-blind manner. Patients in group P received 50 mg rectal diclofenac sodium, 10 micrograms/kg intravenous 0.1% butorphanol tartrate, and 1% lidocaine solution containing 10 micrograms/mL epinephrine for regional anesthesia and for bilateral inferior alveolar nerve blocks before the start of surgery. Postoperative pain intensity at rest (POPI) was assessed on a numerical rating score (NRS) in the postanesthesia care unit (PACU) and on a visual analogue scale (VAS) at the first water intake (FWI) and at 24, 48, and 72 hours after extubation. POPI in the PACU was significantly lower in group P than in group C, whereas there were no significant differences at FWI, 24, 48, and 72 hours after extubation in both groups. Preemptive multimodal analgesia was not observed in this study. PMID- 11432177 TI - Clinical evaluation of inferior alveolar nerve block by injection into the pterygomandibular space anterior to the mandibular foramen. AB - The conventional inferior alveolar nerve block (conventional technique) has potential risks of neural and vascular injuries. We studied a method of inferior alveolar nerve block by injecting a local anesthetic solution into the pterygomandibular space anterior to the mandibular foramen (anterior technique) with the purpose of avoiding such complications. The insertion angle of the anterior technique and the estimation of anesthesia in the anterior technique were examined. The predicted insertion angle measured on computed tomographic images was 60.1 +/- 7.1 degrees from the median, with the syringe end lying on the contralateral mandibular first molar, and the insertion depth was approximately 10 mm. We applied the anterior technique to 100 patients for mandibular molar extraction and assessed the anesthetic effects. A success rate of 74% was obtained. This is similar to that reported for the conventional technique but without the accompanying risks for inferior alveolar neural and vascular complications. PMID- 11432178 TI - Inferior alveolar nerve block by injection into the pterygomandibular space anterior to the mandibular foramen: radiographic study of local anesthetic spread in the pterygomandibular space. AB - We studied the spread of local anesthetic solution in the inferior alveolar nerve block by the injection of local anesthetic solution into the pterygomandibular space anterior to the mandibular foramen (anterior technique). Seventeen volunteers were injected with 1.8 mL of a mixture containing lidocaine and contrast medium utilizing the anterior technique. The course of spread was traced by fluoroscopy in the sagittal plane, and the distribution area was evaluated by lateral cephalograms and horizontal computed tomography. The results indicate that the contrast medium mixture spreads rapidly in the pterygomandibular space to the inferior alveolar nerve in the subjects who exhibited inferior alveolar nerve block effect. We concluded that the anesthetic effect due to the anterior technique was produced by the rapid distribution of anesthetic solution in the pterygomandibular space toward the mandibular foramen, and individual differences in the time of onset of analgesia may be due to differences in the histologic perineural tissues. PMID- 11432179 TI - Adverse reactions triggered by dental local anesthetics: a clinical survey. AB - One hundred and seventy-nine patients completed a questionnaire focusing on adverse reactions to dental local anesthetics as manifested by 16 signs and symptoms. Twenty-six percent of the participants reported having at least 1 adverse reaction. It was found that most of the adverse reactions occurred within the first 2 hours following the injection of local anesthetics. Pallor, palpitations, diaphoresis, and dizziness were the most common adverse reactions reported in the study. The results pointed to a significant relationship between anxiety, gender, injection technique, and procedure with a higher incidence of adverse reactions. PMID- 11432180 TI - Local anesthesia in the palate: a comparison of techniques and solutions. AB - It was the purpose of the present investigation to determine if there were differences in soft-tissue anesthesia in the palate following infiltration and greater palatine nerve block anesthesia and to compare lidocaine with lidocaine plus epinephrine as palatal soft tissue anesthetics. Two studies using 10 volunteers were performed. In one trial, volunteers received a palatal infiltration opposite the second maxillary bicuspid on one side and a greater palatine nerve block on the other. Response to sharp probing and pain-pressure thresholds were measured on each side over a 1-hour census period. In the second trial, volunteers received 2% plain lidocaine as a palatal infiltration on one side and a similar infiltration of 2% lidocaine with 1:80,000 epinephrine on the other in a double-blind randomized fashion. Response to sharp probing was assessed over a 55-minute period. Data were analyzed using Student's paired t tests. The response to sharp probing and pressure-pain thresholds did not differ between palatal infiltration and greater palatine nerve block over the 1-hour period. Lidocaine with epinephrine provided longer lasting anesthesia than plain lidocaine following palatal infiltration (P < .001). Greater palatine nerve block and palatal infiltration provide similar soft-tissue anesthesia. Lidocaine with epinephrine produces longer-lasting soft-tissue anesthesia than plain lidocaine following palatal infiltration. PMID- 11432181 TI - The effects of cigarette smoking on anesthesia. AB - Cigarette smoke contains over 4000 substances, some of which are harmful to the smoker. Some constituents cause cardiovascular problems, increasing the blood pressure, heart rate, and the systemic vascular resistance. Some cause respiratory problems, interfering with oxygen uptake, transport, and delivery. Further, some interfere with respiratory function both during and after anesthesia. Some also interfere with drug metabolism. Various effects on muscle relaxants have been reported. Risk of aspiration is similar to that of nonsmokers, but the incidence of postoperative nausea and vomiting appears to be less in smokers than in nonsmokers. Even passive smoking effects anesthesia. Best is to stop smoking for at least 8 weeks prior to surgery or, if not, at least for 24 hours before surgery. Anxiolytic premedication with smooth, deep anesthesia should prevent most problems. Monitoring may be difficult due to incorrect readings on pulse oximeters and higher arterial to end tidal carbon dioxide differences. In the recovery period, smokers will need oxygen therapy and more analgesics. It is time that anesthesiologists played a stronger role in advising smokers to stop smoking. PMID- 11432182 TI - Anesthetic considerations for orthognathic surgery with evaluation of difficult intubation and technique for hypotensive anesthesia. AB - Orthognathic surgery is carried out to improve facial appearance and/or to improve malocclusion. Usually, patients are young and healthy. However, they may have airway problems. Reinforced silicone low-pressure, high-volume endotracheal tubes and p-xylometazoline (Otrivin) for nasal vasoconstriction reduces problems due to the endotracheal tubes. A head-up position with ventilator and monitoring equipment at the foot end helps the surgeons as well as the surgery. Surgeons may be the cause of endotracheal tube problems. Bleeding is a major problem that may be encountered and is reduced by induced hypotension. During osteotomies, severe bradycardia may occur and may even lead to cardiac arrest. In the early postoperative period, bleeding may be a problem. Later ulceration at the tip of the nose and on the buttocks may be seen if preventive measures are not carried out. PMID- 11432183 TI - Shrinking the dead zone. PMID- 11432184 TI - Sailing on sunlight. PMID- 11432185 TI - A votre sante. Should physicians tell some nondrinkers to start? PMID- 11432186 TI - Fuel cell phones. PMID- 11432187 TI - Sigma chi chimpy. PMID- 11432188 TI - Napoleon's revenge. In the U.S., height hits its head on the genetic ceiling. PMID- 11432189 TI - In a dry land. The southwest faces a dry future, but there are ways to cope. PMID- 11432190 TI - Builders of light pipes. PMID- 11432193 TI - How to build a hyper computer. PMID- 11432192 TI - Starbucks in the forbidden city. PMID- 11432194 TI - The truth and the hype of hypnosis. PMID- 11432195 TI - Making molecules into motors. PMID- 11432196 TI - Frozen light. PMID- 11432197 TI - Battling biofilms. PMID- 11432198 TI - Fishy business. PMID- 11432199 TI - Tan or burn. PMID- 11432200 TI - The universe atop a mountain. PMID- 11432201 TI - How can sea mammals drink saltwater? PMID- 11432202 TI - Toll-like receptors and innate immunity. PMID- 11432203 TI - Attractions and migrations of lymphoid cells in the organization of humoral immune responses. PMID- 11432204 TI - Factors and forces controlling V(D)J recombination. PMID- 11432205 TI - T cell effector subsets: extending the Th1/Th2 paradigm. PMID- 11432206 TI - MHC-restricted T cell responses against posttranslationally modified peptide antigens. PMID- 11432207 TI - Gastrointestinal eosinophils in health and disease. PMID- 11432208 TI - Chemokines in immunity. AB - Chemokines are a superfamily of small, heparin-binding cytokines that induce directed migration of various types of leukocytes through interactions with a group of seven-transmembrane G protein-coupled receptors. At present, over 40 members have been identified in humans. Until a few years ago, chemokines were mainly known as potent attractants for leukocytes such as neutrophils and monocytes, and were thus mostly regarded as the mediators of acute and chronic inflammatory responses. They had highly complex ligand-receptor relationships and their genes were regularly mapped on chromosomes 4 and 17 in humans. Recently, novel chemokines have been identified in rapid succession, mostly through application of bioinformatics on expressed sequence tag databases. A number of surprises have followed the identification of novel chemokines. They are constitutively expressed in lymphoid and other tissues with individually characteristic patterns. Most of them turned out to be highly specific for lymphocytes and dendritic cells. They have much simpler ligand-receptor relationships, and their genes are mapped to chromosomal loci different from the traditional chemokine gene clusters. Thus, the emerging chemokines are functionally and genetically quite different from the classical "inflammatory chemokines" and may be classified as "immune (system) chemokines" because of their profound importance in the genesis, homeostasis and function of the immune system. The emergence of immune chemokines has brought about a great deal of impact on the current immunological research, leading us to a better understanding on the fine traffic regulation of lymphocytes and dendritic cells. The immune chemokines and their receptors are also likely to be important future targets for therapeutic intervention of our immune responses. PMID- 11432209 TI - A final lesson. PMID- 11432210 TI - End-of-life issues in intensive care units: a national random survey of nurses' knowledge and beliefs. AB - OBJECTIVE: To investigate the knowledge, beliefs, and ethical concerns of nurses caring for patients dying in intensive care units. METHODS: A survey was mailed to 3000 members of the American Association of Critical-Care Nurses. The survey contained various scenarios depicting end-of-life actions for patients: pain management, withholding or withdrawing life support, assisted suicide, and voluntary and nonvoluntary euthanasia. RESULTS: Most of the respondents (N = 906) correctly identified the distinctions among the end-of-life actions depicted in the scenarios. Almost all (99%-100%) agreed with the actions of pain management and withholding or withdrawing life support. A total of 83% disagreed with assisted suicide, 95% disagreed with voluntary euthanasia, and 89% to 98% disagreed with nonvoluntary euthanasia. Most (78%) thought that dying patients frequently (31%) or sometimes (47%) received inadequate pain medicine, and almost all agreed with the double-effect principle. Communication between nurses and physicians was generally effective, but unit-level conferences that focused on grief counseling and debriefing staff rarely (38%) or never (49%) occurred. Among the respondents, 37% had been asked to assist in hastening a patient's death. Although 59% reported that they seldom acted against their consciences in caring for dying patients, 34% indicated that they sometimes had acted against their conscience, and 6% had done so to a great extent. CONCLUSIONS: Intensive care unit nurses strongly support good pain management for dying patients and withholding or withdrawing life-sustaining therapies to allow unavoidable death. The vast majority oppose assisted suicide and euthanasia. Wider professional and public dialogue on end-of-life care in intensive care units is warranted. PMID- 11432211 TI - End-of-life care in the intensive care unit: a challenge for nurses. AB - Results from several research studies combined with increasing public tensions surrounding physician-assisted suicide have fueled a growing awareness of the inadequacies of end-of-life care. Investigators also suggest that intensive care unit nurses have a limited role in end-of-life decision making and care planning. This article explores cultural issues influencing end-of-life care in intensive care units, explores factors surrounding the limited involvement of critical care nurses in end-of-life decision making and care planning, and offers recommendations for changing nursing practice. Because improving end-of-life care will require cultural changes, an understanding of the cultural issues involved is needed. Recommendations for changing nursing practice include a model of end of-life care that incorporates the goals of both cure and comfort care, as well as a shared decision-making process. Nurses are essential to improving end-of life care in today's intensive care units. PMID- 11432212 TI - Patients' perceptions and responses to procedural pain: results from Thunder Project II. AB - BACKGROUND: Little is known about the painfulness of procedures commonly performed in acute and critical care settings. OBJECTIVE: To describe pain associated with turning, wound drain removal, tracheal suctioning, femoral catheter removal, placement of a central venous catheter, and nonburn wound dressing change and frequency of use of analgesics during procedures. METHODS: A comparative, descriptive design was used. Numeric rating scales were used to measure pain intensity and procedural distress; word lists, to measure pain quality. RESULTS: Data were obtained from 6201 patients: 176 younger than 18 years and 5957 adults. Mean pain intensity scores for turning and tracheal suctioning were 2.80 and 3.00, respectively (scale, 0-5), for 4- to 7-year-olds and 52.0 and 28.1 (scale, 0-100) for 8- to 12-year-olds. For adolescents, mean pain intensity scores for wound dressing change, turning, tracheal suctioning, and wound drain removal were 5 to 7 (scale, 0-10); mean procedural distress scores were 4.83 to 6.00 (scale, 0-10). In adults, mean pain intensity scores for all procedures were 2.65 to 4.93 (scale, 0-10); mean procedural distress scores were 1.89 to 3.47 (scale, 0-10). The most painful and distressing procedures were turning for adults and wound care for adolescents. Procedural pain was often described as sharp, stinging, stabbing, shooting, and awful. Less than 20% of patients received opiates before procedures. CONCLUSIONS: Procedural pain varies considerably and is procedure specific. Because procedures are performed so often, more individualized attention to preparation for and control of procedural pain is warranted. PMID- 11432213 TI - Pain experiences of traumatically injured patients in a critical care setting. AB - BACKGROUND: Little is known about the acute pain experiences of traumatically injured critically ill patients. OBJECTIVES: To describe pain experiences of traumatically injured adults during the first 72 hours of hospitalization. METHODS: Thirty multiply injured adults at a level I trauma center participated in the study. Pain was measured by using the McGill Pain Questionnaire and a visual analog scale. Subjects completed pain measures while at rest in a supine recumbent position and after a turn onto the side. RESULTS: The typical subject was 37 years old, had 4 major blunt trauma injuries, and had received the equivalent of 55.9 mg of morphine during the 24 hours before data collection. Mean at-rest scores were 26.5 on the pain-rating index, 2 on the present pain intensity index, and 34.6 on the visual analog scale. Immediately after the turn, mean scores on the visual analog scale increased from 25 to 48.1 (P = .002). Other pain scores after the turn did not differ significantly from at-rest values. Subjects who turned had lower scores on the visual analog scale at rest (P = .02) and less anxiety (P = .02) than did those who refused to turn. Ninety six percent reported pain in the injured areas, and 36% reported pain related to biomedical devices. No relationship was found among reported pain and demographic, treatment, or clinical variables. CONCLUSIONS: Additional research is needed on pain at rest and during commonly performed procedures and on improved methods for pain relief in traumatically injured critically ill patients. PMID- 11432214 TI - Electrophysiology study: a comprehensive review. AB - An EPS provides valuable information about cardiac arrhythmias by recording intracardiac electrograms and using programmed stimulation techniques. Although the sequence of study protocols may vary with each institution, an EPS routinely begins with baseline measurement of conduction intervals. Subsequently, clinical arrhythmias are induced either by incremental or decremental pacing and extrastimulus techniques. Nurses must understand the technical and psychosocial aspects of an EPS to ensure adequate care. The primary role of nurses is to maintain a safe environment for the patient in the electrophysiology laboratory (i.e., maintain aseptic techniques and intervene when complications occur). Rapid technological advances and the increasing number of well-educated patients require that nurses be knowledgeable and skilled in providing progressive care, such as developing educational programs and support groups for patients and patients' families. PMID- 11432215 TI - Comparison of fever treatments in the critically ill: a pilot study. AB - BACKGROUND: Fever in critically ill patients is often treated with antipyretics or physical cooling methods. Although fever is a host defense response that may benefit some critically ill patients, others may not tolerate the cardiovascular demands associated with fever. OBJECTIVES: To compare antipyretics and physical cooling for their effects on core body temperature and cardiovascular responses in critically ill patients. METHODS: The antipyretic administered was 650 mg of acetaminophen. Physical cooling was accomplished by anterior placement of a cooling blanket at 18 degrees C. Core temperature and cardiovascular responses were measured in 14 febrile (body temperature, 38.8 degrees C) critically ill patients at baseline before treatment and up to 3 hours after treatment. Patients able to receive acetaminophen were randomly assigned to receive either acetaminophen only (n = 5) or acetaminophen in combination with a cooling blanket (n = 3). Patients not able to receive acetaminophen were treated with physical cooling only (n = 6). RESULTS: Mean body temperature decreased minimally from baseline to 3 hours after treatment in the physical-cooling-only group (from 39.1 degrees C to 39.0 degrees C) and in the physical cooling and acetaminophen group (from 39.1 degrees C to 38.6 degrees C), but the mean body temperature increased in the acetaminophen-only group (from 39.2 degrees C to 39.4 degrees C). Other notable findings included a slight increase in systemic vascular resistance index in the physical-cooling-only group and in the physical-cooling-plus-acetaminophen group. CONCLUSIONS: Although the study included only 14 subjects, the findings will provide information for future studies in febrile critically ill patients. PMID- 11432216 TI - Creating a culture of safety and improvement: a key to reducing medical error. PMID- 11432217 TI - Heart failure: update on treatment and prognosis. AB - HF is a prevalent and debilitating disease, affecting nearly 5 million patients and perhaps an equal number with asymptomatic left ventricular dysfunction who are at high risk of atrial fibrillation developing. An estimated 550,000 new cases occur every year. HF is the most common diagnosis in hospitalized patients aged 65 and over and is a major cause of death. The median survival after onset is 1.7 years in men and 3.2 years in women. The majority of cardiac deaths in patients with HF are sudden and arrhythmogenic: the rest are due to progressive hemodynamic deterioration. A significant advance in the past decade has been the recognition of the importance of inhibiting the neurohormonal action in HF with the use of beta-blockers, angiotensin receptor, and aldosterone antagonists. In addition, a new concept in HF therapy has evolved. The view that chronic HF is an irreversible, end-stage process is being supplanted by the fact that it is possible to effect biological improvement in the intrinsic defects of function and structure in hearts afflicted with chronic HF. Reversibility of HF has been reported by (1) unloading the failing heart using an LVAD, (2) the sophisticated use of diuretic combinations and neurohormonal blocking drugs, or (3) employing continuous arteriovenous hemofiltration. Thus it is now possible to reverse a process that has long been considered irreversible. Exercise programs designed for patients with HF that have been advocated recently can be difficult to apply. Fine tuning of an exercise regimen is required because a reduction in cardiac work is mandatory when treating HF, where the concern is that the heart may not be capable of supplying the metabolic needs of the body, even in resting states. Finally, although not emphasized in the recent literature on HF, the use of diuretics and sodium restriction continue to be the mainstays of therapy without which compensation of HF is not possible. PMID- 11432218 TI - One solution to poor staffing ratios. PMID- 11432219 TI - An essay on criticism. PMID- 11432220 TI - Cooling by convection vs cooling by conduction for treatment of fever in critically ill adults. PMID- 11432221 TI - [Pancreatoblastoma in a child (case report)]. AB - This paper presents a rare form of pancreatic malignancy in childhood, pancreatoblastoma, tumor rarely encountered in children and adolescents. This tumor is also very interesting because of differential diagnosis toward other retroperitoneal tumors in children. They are most frequently diagnosed accidentally during clinical or ultrasonographic examination. We are presenting a case of 14 year old girl in whom a tumor was found during the investigation for the dull pain below the left rib margin. Tumor was successfully removed surgically. A 26 months follow-up was without any signs of recurrence of the primary tumor. PMID- 11432222 TI - New aspects in laparoscopic cystic artery anatomy. AB - While performing laparoscopic cholecystectomy we noticed lateral branches of the cystic artery as well as arterial bleeding from the gallbladder bed. In order to establish the nature of these blood vessels we undertook a corrosion cast study. The features analyzed were their frequency of occurrence, caliber and length. The research was performed on 50 human autopsy specimens of the liver with the hepatoduodenal ligament. Lateral branches of the cystic artery that irrigate a part of the anteromedial segment of the right liver lobe were observed in 10% cases. Anastomotic bridges between the cystic artery and the arterial system of the liver were observed in 12% cases. PMID- 11432223 TI - [Reconstruction of the hypopharynx and cervical esophagus using a free jejunal graft]. AB - Extensive malignant tumors of the hypopharynx and cervical esophagus continue to challenge surgeons in respect to both type and extent of resection as well as type of reconstruction. In the period between November 1st, 1996 and November 1st, 1998, at our Department, five patients have been operated due to squamocellular carcinoma of the hypopharynx using a free jejunal graft reconstruction method. The first free jejunal graft operation due to hypopharyngeal carcinoma, at the same time the first operation of this kind ever done in our Country, was performed on November 13th, 1998. There were 4 female and one male patient, average age 47.75 years. Disfagia for solid foods was a leading symptom in all patients (mean duration of 3.5 months) and was always accompanied with weight loss (average of 8 kg for two months). In all patients barium swallow, endoscopy, CT as well as intraoperative endoscopy was performed. Radical surgical procedure was always accompanied with the bilateral modified lympf node neck dissection. As a arterial donor vessel superior thyroid artery was used in all patients. As a venous drainage in three patients a external jugular vein was used and in two facial vein. Reconstruction using a free jejunal graft of approximately 25 cm long was performed in all patients creating upper, oropharingeal, anastomosis end to side and distal, esophageal, end to end (in only one patients side to end) using 3/0 apsorbable sutures. Mean duration of the operation was six hours. The postoperative course in all patients was uneventful. On the 9th postoperative day gastrografin and three days later barium swallow radiography was performed as a standard control study. Regular check ups were done on three, six, nine months, year and two years. On all controls all patients were symptom free and feeling well. It is our opinion that in the patients with isolated carcinoma of the hypopharynx due to low morbidity and mortality rate, free jejunal graft method is the surgical procedure of choice. PMID- 11432224 TI - [Surgical treatment of cardial carcinoma]. AB - In the period from 1970 to 1996, in the Departmenf of Esophagogastric Surgery, Belgrade, Yugoslavia 562 have been operated with curative intent due to cardiac carcinoma. Since 1970 until 1985, in 182 patients a distal esophagectomy and proximal gastrectomy followed by eosphagogastro anastomosis had been performed. In the period between 1982 and 1996, a distal esophagectomy with total gastrectomy and radical or extended radical dissection and intrathoracic esophagojejuno anastomosis in 380 patients (192 hand sewn 3/0 interrupted sutures and 188 spapled anastomosis) had been performed. In our opinion for the majority of patients with advanced cardiac carcinoma distal esophagectomy and total gastrectomy, via the left thoracoabdominal approach, with D2 pancreas preserving splenectomy and dissection of lymph nodes in stations 16a1 and 16a2 is a surgical therapy of choice. Overall complications of such a procedure not differ in type and number from those after standard total gastrectomy and D2 dissection. PMID- 11432225 TI - [Surgical strategy in the treatment of primary lymphoma of the stomach]. AB - Primary gastric lymphoma has been traditionally accounted for only 1-7% of all malignancies of the stomach. During the period 1980-1996 in Department of Esophagogastric Surgery of University Surgical Hospital in Belgrade 41 patients have been operated due to primary gastric lymphoma. Radical surgery is the only therapeutic procedure in more then 45% of patients. It is also a safe procedure with specific complication rate less then 22%, and specific mortality rate less then 5%. In our experience, total gastrectomy with systematic lymphadenectomy is a therapy of choice in most patients with primary gastric lymphoma. PMID- 11432226 TI - [A large duplication cyst of the proximal section of the stomach in an adult- case report]. AB - Intestinal duplication is a rare congenital anomaly. It appears through out entire gastrointestinal tract, specially in the ileum. Gastric duplication cyst in adults is extremely rare. Diagnosis is often missed, often established at operation. Preoperative diagnosis may be suspected on CT or MRI with contrast given oraly as well as on EUS. Management of symptomatic gastric duplications is surgical. Recommended management is complete exscision of the duplication without major violation of the gastric lumen. In most cases giant cyst does not allow smaller resection but partial gastrectomy. In this report a rare case of gastric duplicatione cyst of the proximal stomach is presented. Diagnostic finding and surgical therapy are discussed. PMID- 11432227 TI - [Localized plano-cellular type of Castleman's disease]. AB - Castleman was first to study lymphoid tumour of the mediastinum with characteristic histologic features which had until then been mistaken for thymommas grossly, radiologically and histologically. It was soon evidented that the condition occurred in regions other then the mediastinum and even in places where lymph nodes are not normally presents. Localised plasma cell type is rare variant of Castlemans disease, especially as an intraabdominal variant. Most patients present with local symptoms, but general malaise or fever, anaemia, elevated sedimentation rate, polyclonal hypergamaglobulinemia etc. can be present. Using preoperative imaging procedures solid, well-circumscribed expansive mass can be detected. After surgical excision and patohistological examination diagnosis can be established. In this paper we report rare case of localised plasma cell type of Castlemans disease. Other types of disease, diagnostic findings and therapeutic possibilities are discussed. PMID- 11432228 TI - [Endoscopic surgery of the esophagus]. PMID- 11432229 TI - Significance of duodenogastric reflux in patients with erosive esophagitis. AB - The role of duodenogastric reflux (DGR) in producing esophageal mucosal injury across the spectrum of GERD is still controversial. Our objective was to assess the role of DGR in the genesis and evolution of erosive esophagitis. Forty patients are presented who meet the criteria for the diagnosis of erosive esophagitis. Symptom scoring, endoscopy and mucosal biopsy with patohistologic classification of erosive esophagitis and a quantification of DGR, using scintigraphic imaging after intravenous injection of 99mTc-labeled HIDA, was performed. DGR was demonstrated in half of the patients. Both groups of patients (with and without DGR) were treated with life style changes, cisapride and nizatidine for a period of six weeks. After this period, symptom scoring, endoscopy and biopsy were performed again in both groups. There has been a significant decrease in the level of symptom scoring and endoscopic inflammation after the therapy, but without statistically significant difference between these two groups. Patohistologic finding after the medical treatment showed a marked difference between the two groups concerning the presence of DGR, indicating that the level of inflammation is statistically significantly higher in the group with DGR. This study showed that even if symptom scoring and endoscopy revealed improvement after the therapy in both groups of patients, patohistologic mucosal damage in patients with erosive esophagitis is greatly dependent of the presence of DGR. We conclude that alkaline component of GERD should be considered in the evaluation of patients with erosive esophagitis so that appropriate surgical therapy can be instituted. PMID- 11432230 TI - Laparoscopic cardiomyotomy in the treatment of esophageal achalasia. AB - Tu evaluate the usefulness of the laparoscopic approach as the standard procedure in the surgical treatment of achalasia. Among different competing options of the treatment of esophageal achalasia, extramucosal myotomy of the lower esophageal sphincter--usually combined with anterior fundoplasty--is the most effective but also the most invasive approach. Minimally invasive performance of this operation reduces invasivity and should make the operative treatment a more attractive alternative to other procedures, such as pneumatic dilatation or botox injection. From 1991 till 1997, 27 patients underwent laparoscopic Heller Dor operation (16 males, 11 females, mean age 37 years). Diagnosis was established in all of them by an esophagogram and esophageal manometry. The main symptom was dysphagia in all of the patients. No mortality was observed in this series. There were no conversions to laparotomy. The single intraoperative complication was one case of iatrogenic mucosal laceration. Post operative complications were found in one case of wound infection, and two cases of pneumomedistinum. After a mean follow up of 33 months (3-77), all patients are without dysphagia and without pathological gastroesophageal reflux. The mean value of residual LES pressure could be reduced from 21 +/- 6.4 mmHg to 7.44 +/- 2.7 mmHg. Laparoscopic cardiomyotomy is at lesat as safe, in terms of morbidity and mortality, as open surgery and similarily effective in alleviating dysphagia. Short hospitalisation and convalascent periods have provided an attractive alternative to repeated dilations for many patients. PMID- 11432231 TI - [Myocardial revascularization using the internal thoracic artery: pro and con]. AB - Internal thoracic artery (ITA) has superior histological, physiological and pharmacological properties over the venous grafts, and it is considered to be the graft of choice for myocardial revascularization. It has low incidence of late atherosclerotic lesions, and excellent long-term patency. Usage of ITA yields improved clinical results in all subgroups of patients with coronary artery disease, including patients with poor left ventricular function (EF%), left main stenosis, diffuse coronary artery disease and octogenarians. Since superior long term results (to be expected only 10-15 years after the operation) may be outweighed with slightly higher incidence of early morbidity and mortality, many cardiac surgeons are reluctant to use it routinely. However, this attitude is difficult to justify, since early operative results in patients that have received ITA graft are superior as well, compared to patients that have received vein grafts only. PMID- 11432232 TI - [Acute lung injury related to blood transfusion]. AB - Transfusion-related acute lung injury (TRALI) is a second most serious complication of the blood transfusion. It is a group of symptoms and signs such as dyspnoea, hypotension, cyanosis, cough, elevated temperature, fever and lung oedema that usually develops within an hour or two after transfusion. The full stage clinical presentation is developed between 4th and 6th hours after transfusion. The syndrome is caused by leucoagglutinins or by other lymphocytotoxic antibodies specific for some antigens present on the donor's leukocytes. Alveoles of the lung are the main place of the pathological changes such as intra-alveolar oedema, haemorrhage, hyaline membrane formation, alveolar cell hypertrophy and scant interstitial inflammation. Chest X-ray showed bilateral pulmonary infiltrates but without vascular congestion and with normal cardiac silhouette comparing to the status before transfusion. The syndrome has to be distinquished from pulmonary oedema caused by acute cardial insufficiency, overhydration, trauma and sepsis. PMID- 11432233 TI - [Overview of reconstructive surgery and functional compensation in the digestive tract]. AB - This paper represents a short and overall overview of the reconstructive procedures of the esophagus, stomach, colon and rectum. Optimal reconstructive procedure has not been chosen only for the reconstruction of the digestive system. Precise selection of the patients and reconstructive methods must enable the correction of the possible functional disadvantages. Usage of adaptional and reserve possibilities of other organs and tissues will help the functional compensation of the lost segments of the digestive system. The understanding of the importance of the physiological approach to the reconstruction of the digestive system using the segments of stomach, jejunum or colon is of the most importance so that in this difficult segment of surgery adequate improvements can be achieved. PMID- 11432234 TI - [A new concept of surgical treatment of uterine and vaginal prolapse]. PMID- 11432235 TI - Rectal cancer in the year 2000. AB - The last twenty years have seen significant advances in quality of surgery for rectal cancer. More sphincter saving procedures are being carried out. The variance in local recurrence rate between surgeons has been recognised and this has stimulated a more uniform approach to dissection involving removal of the mesorectum. The place of radiotherapy is rapidly being defined. Future developments are very likely to be based upon the present. Thus further work in screening to achieve earlier diagnosis is likely to be extended to the genetic identification of patients at risk with follow up for these selected individuals. Education of the general population is already improving and will continue to do so. Pre-operative staging by imaging will become more accurate allowing rational adoption of management policies before histological examination of any resected specimen. This will achieve refinement of treatment combinations. Finally it is likely that new and effective chemotherapeutic agents will be developed. PMID- 11432236 TI - Total mesorectal exsicion (TME). AB - For almost twenty years the concept of total mesorectal excision (TME) has featured in the medical literature. It remains however the object of ongoing controversy and sometimes of downright antagonism. At the present time there are at least four other names for exactly the same technique being championed by different authorities. 1. Circumferential Mesorectal Excision (Enker); 2. Sharp Mesorectal Excision (Cohen); 3. Extrafascial Excision of the Rectum (Hill); 4. Total Anatomical Dissection (Boley). As Shakespeare said "A rose by any other name would smell as sweet". PMID- 11432237 TI - Long-term results of cutting seton fistulotomy. AB - BACKGROUND/AIMS: To determine the long term outcome of cutting seton fistulotomy. MATERIALS AND METHOD: An audit of fistula surgery using a cutting seton for cryptoglandular sepsis identified 32 patients treated between 1988-1996. The majority of the patients had a trans-sphincteric (26/32, 81%) fistula. RESULTS: Fourteen (44%) had had previous fistula surgery before seton fistulotomy. Follow up was complete in 28 patients: 8 patients had recurrence (29%), 4 at a new site and 4 at the same site after apparent healing. Major incontinence was reported by 3 patients, minor incontinence by 7 and soiling by 5, but only 7 patients worsened after the cutting seton fistulotomy. Major incontinence was reported only by women who had a previous vaginal delivery. CONCLUSIONS: Cutting setons do not always preserve continence in high trans-sphincteric and complex fistulas. Patients should be warned about a high recurrence rate after cutting seton fistulotomy. The use of a cutting seton for anterior fistulas in female patients with a history of vaginal delivery should be avoided because of the risk of incontinence. PMID- 11432238 TI - Hereditary colorectal cancer syndromes. AB - Tumors of large bowel continue to be one of the leading causes of morbidity and mortality with about 300,000 new cases and 200,000 deaths per year in Europe and USA despite recent technological advancements. In sharp contrast with these discouraging data, the basic knowledge of colorectal neoplasms has grown remarkably in the last decades especially with the genetic elucidation of the two inherited cancer-predisposition syndromes, familial polyposis (FAP) and hereditary non polyposis colorectal cancer (HNPCC). Recognition of the genetic component of CRC is growing; gene mutations responsible for cell transformation can be present as inherited germline defect or arise in somatic cells as consequence of environmental insults. The two main hereditary syndromes, FAP and HNPCC, account for about 6-10% of CRCs, remaining cases are attributed to so called sporadic cancer. Although the timescale of the appearance and risk of recurrence of the hereditary and sporadic forms are quite different, they share a common pathway: the adenoma to carcinoma sequence. In 1990 Fearon and Vogelstein proposed a multistep model for the molecular events underlying colorectal tumorigenesis. The model was based on two assumptions: the first one is that the tumors are clonal, the second assumption is that the colorectal tumorigenesis occurs as succession of a series of events that can be described as dyplasia carcinoma sequence or adenoma-carcinoma sequence. The initial alterations which are not detectable on histologic examination, are subtle changes in the normal balance between cell growth and cell death. With progression precursors to adenoma, the foci of aberrant cripts become detectable. Few adenomas progress to carcinoma, however if the progression of these lesions remain unchecked, there is an increased risk of tumor diffusion. As the cells need time to accumulate the genetic defects including mutational activation of oncogenes and inactivation of tumor suppressor genes to undergo full malignant transformation, CRC occurs mainly in the elderly. If one of these defects are present at birth as germline mutations, fewer mutational events will be requested to reach malignant transformation and the disease will appear earlier(). PMID- 11432239 TI - The final diagnosis in pouch patients for presumed ulcerative colitis may change to Crohn's disease: patients should be warned of the consequences. AB - BACKGROUND: The ileal pouch is usually the procedure of choice in the surgical management of Ulcerative Colitis (UC), but pouch surgery has a high complication and failure rate in Crohn's Disease (CD). METHODS: This study is a review of 222 consecutive patients with an initial diagnosis of UC and 3 with CD, treated by ileal pouch anal anastomosis. RESULTS: Only 3 of 225 patients treated by ileal pouch anal anastomosis had a preoperative diagnosis of CD. At the time of follow up the diagnosis was 23 CD and 202 UC (median follow-up 118.6 months for CD and 91.3 months for UC). 10 of 23 patients with CD had an 1 stage proctocolectomy and pouch and 13 had an initial colectomy. The following were significantly more common in CD than UC: ileo-anal sepsis (30% v 12%: p = 0.028) and pouchitis (65% v 32%: p = 0.0016). The pouch excision rate was 47.8% in CD and 10.9% in UC (p). Uniquely we found that 8/11 patients with CD had perineal sinus after pouch excision compared with only 1/22 in those with UC (p). The median functional score (scale 0-12), in those with intact pouches, was the same in CD and UC: 4 and 3 respectively. CONCLUSION: Our findings show that ileal pouch anal anastomosis is a satisfactory procedure for UC but that the diagnosis had to be changed to CD in 20 of 222 cases. Thus patients should be warned about a 48% failure rate and a high incidence of complications if CD declares itself during follow-up but Crohn's patients who retain their pouch have functional results that are indistinguishable from UC. PMID- 11432240 TI - Long-term results after low anterior stapled anastomosis. AB - In the period 01.01.1991-12.31.1996, 286 low anterior stapled resections of the rectum due to rectal carcinoma were performed at the First Surgical Clinic, the Third Department for Colorectal Surgery, Belgrade. There were 57% males and 43% females, median age 59.6 years. The most common localization of tumor was in the distal third of the rectum 181 (63%). In the middle third, there were 89 (31%) and in the upper, intraperitoneal third 16 (6%). Histopathological examination revealed adenocarcinoma in all cases. All operative specimens were examined by one pathologist and classified according to the Gunderson-Sosin modification of Dukes classification. There were 14 (4.9%) in stage A, 167 (58.4%) in stage B (B1,B2,B3), 89 (31.1%) stage C (C1,C2,C3) and 16 (5.6%) stage D. According to Broders classification, there were 129 (45%) well differentiated, 142 (50%) moderately and 15 (5%) poorly differentiated tumors. Anastomotic dehiscence was found in 17 patients (5.95%), mostly conservatively treated, except in 6 cases where spreading peritonitis developed requiring operative treatment. There were 9 (3.1%) postoperative deaths, a half of them with specifically operation related mortality. Recurrence of the disease was registered in 47 (18%) patients, out of 260 who were regularly followed up (26 were lost). Local recurrence alone was found in 21 (8.0%) patients, while distant spread was registered in 22 (8.46%) patients. Local and distant spread was found in 4 (1.5%) cases. At present, the median follow-up is at 54 months. Analysis by the Kaplan-Meier's test shows cumulative survival of 66% at 71 months of the follow-up. Seventy four percent of patients with curative operations exhibit no signs of recurrence at 5 years. PMID- 11432241 TI - The role of overlapping sphincteroplasty in traumatic fecal incontinence. AB - Fecal incontinence is one of the most distressing conditions. Even there is a great variety of etiologic factors, they can be systematized in two main categories; fecal incontinence with normal and fecal incontinence with abnormal function of pelvic floor muscles. The latter is more interesting for surgeons since this category includes the great majority of surgically caused and surgically correctable fecal incontinences. Disruption of the anal sphincter caused by obstetric injury, anorectal operations or external trauma is the commonest cause of fecal incontinence. In the period 1990-1999, 53 patients with fecal incontinence caused by sphincter injury were treated on the third department for colorectal surgery, First Surgical Clinic. There were 43 females and 10 males with a mean age of 36.1 years (range 18-64). Causes of fecal incontinence were: obstetric trauma 38 patients. (71%), fistulotomy 9 (17%), war injuries 3 (6%) and nonspecific 3 (6%). The severity of incontinence was graded by Browning--Parks's classification. There were no patients in group A and B, in group C were 11 patients and in D group 37 patients. Wexner score system was also utilized preoperatively and postoperatively to determine continence function more precisely. Clinical exam, anoscopy and special investigations, such as anal manometry, EMG and defecography were carried out in all cases. Period between injury and repair was between 6 months and 20 years. Severe pudendal neuropathy was present in 17 patients. Five patients had oostomies performed at the time of injury. In three cases was present traumatic cloaca and in one case rectovaginal fistula. Overlapping sphincter repair technique was carried out in all cases. Full bowel preparation and antibiotics were prescribed. Anterior sphincteroplasty was performed in 39 cases, lateral in 7, posterior in 3 and anterior sphincteroplasty + posterior plication of puborectalis (Parks operation) in 4 patients. Protective colostomy was not performed in any case. The outcome of the procedure was considered as excellent, good, fair and poor (excellent when full control of solid and liquid stool and flatus was achieved, good when there was continence to feces but not to flatus, fair when patients could control only solid feces and poor when only partial control of solid feces was obtained). The overall functional results were as follows: Excellent in 25 patients (47.2%), good in 12 (22.7%), fair in 11 (20.7%) and poor in 5 (9.4%). Wound infection occurred in 9 (16.9%) patients, leading to disruption of sutures in three patients with consequent poor results. Two of them were reoperated. The duration of follow-up was from 5 to 60 months. We conclude that an overlapping sphincteroplasty is a method of choice in treating fecal incontinence caused by trauma. The best results are achieved with anterior sphincteroplasty. Results of posterior sphincteroplasty were disappointing. If present, pudendal neuropathy directly influences the outcome of sphincteroplasty. Manometric studies correlate with the clinical outcome. PMID- 11432242 TI - Medical management of inflamatory bowel diseases--therapeutic options. AB - Corticosteroids and the 5-aminosalicylate (5-ASA) drugs still constitute the mainstays of therapy, with azatioprine/mercaptopurine an established second-line agent for resistant disease. Primarily nutritional therapy has a place for some patients with Crohn's disease, and alternative immunosupressants are finding their own disease. The medical management of inflammatory bowel diseases is poised to enter to new era if the current promise shown by investigatory immunomodulatory regimes translates into confirmed effective therapy. PMID- 11432243 TI - [Defecography study in disorders of the rectum and anal canal]. PMID- 11432244 TI - [Presacral cysts, diagnosis, surgical treatment]. PMID- 11432245 TI - [Surgical treatment of chronic anal fissure, how and why]. PMID- 11432246 TI - Postoperative pathological examination of colorectal cancer. AB - The close cooperation of the surgeon and the pathologist in clinical practice and research of colorectal cancer was far established. The histopa-histopathological report has to include all important information in relation to tumour classification, prognostic factors and surgical procedure. This is the starting point for further treatment, estimation of prognosis and treatment results, indicator of oncological quality of surgical procedure and the most important contribution to increasing knowledge of clinical pathobiology related to colorectal cancer. To reach these goals the pathologist should follow some of up dated histopathological protocols for the examination of specimen removed from patients with colorectal carcinoma. Most of the proposals and international standards are presented in detail. The applying of three main classifications for colorectal carcinoma is mandatory, i.e. histopathological classification, the disease stage (pathoanatomic extension) classification and residual disease classification. It seems that the first and most important step is R (residual disease) classification, serving as the most reliable determinant in further treatment, prognosis and in assessment of surgical treatment. The focus of surgery on R0 category subgroup of patients is presented in relation to the disease stage as second proven prognostic factor, strongly influencing the prognosis and adjuvant therapy options. The detection and significance of minimal residual disease, mainly occult micrometastases in examined lymph nodes are essential for more accurate staging of the disease. Our own data from pilot study confirmed the significant stage migration (Will Rogers phenomenon of "up grading"). Using combination of serial sectioning and immunohistochemical reactivity of anti-cytokeratin antibody to scattered micrometastatic foci in lymph nodes and perirectal fat, we have shown the presence of minimal residual disease in 23.53% of R0 pT3 pN0 primary classified cases. Correlation to other probable and possible histological independent prognostic factors is discussed, as well as the most significant molecular prognostic markers. PMID- 11432247 TI - Coloanal anastomosis in surgery for rectal cancer. AB - The definition of coloanal anastomosis should be based on anatomy of the anal canal. The anal canal is terminal portion of the digestive tract and comprises muscle tube of approximately 3 cm long with a lumen coated with several types of mucosa or epithelium. Muscle tube of the anal canal consists of two layers, inner sphincter muscle and external sphincter muscle. The former is a continuation of rectal muscle layer and the latter extends to the levator ani muscle. Colo-anal anastomosis (CACA): whole of the rectum is excised with a separation line placed just above the anal canal. An anastomosis is made between colonic cut end and upper limit of the anal canal. Colo-anus anastomosis in a narrow sense (CAA): rectum is excised with a bit of upper part of the anal canal. Then an anastomosis is made inside the anal canal. PMID- 11432248 TI - [Correction of failed primary surgical interventions in benign lesions of the esophagus]. AB - In an retrospective analysis of documentation and printed paper in period between 1964-1998 168 patients required reoperations in benign esophageal lesions. Failed initial interventions were found in 66 (39.28%) pts. operated in our clinic and 102 (60.71%) pts operated in other institutions in our country. The corrective procedure was possible in 140 (83.33%) pts and impossible in 11 (6.54%). Post operative mortality was 7.73% so the definitive failed corrections were 14.27%. The main cause of failure were: Defective operative technique, wrong preoperative diagnosis and indications including inadequate initial procedure. Among the 168 pts undergoing corrective surgery, more than a half of pts had obstructive lesions requiring resections and reconstructions. Dilatation of the stricture was possible in 16% followed by total diversion, particularly in previous gastric surgery. Malignant degeneration of benign lesions were found in 10.71%. Only 4 pts (2.3%) refused suggested operations. Satisfactory long-term results especially in patients undergoing esophageal corrective reconstructions for benign diseases were good to excellent in regular yearly postoperative routine control examination. PMID- 11432249 TI - [Study of blood flow in liver hemangiomas using radionuclide angiography]. AB - Hepatic radionuclide angiography (HRA) is a recognised method of investigation of liver blood flow disorders caused by: diffuse and focal diseases of liver parenchyma or disorders of blood flow in extrahepatic liver vessels. Hepatic perfusion index (HPI) based on Sarper's slope method is significantly lower in patients with e.g. liver cirrhosis, malignant primary and metastatic liver diseases and portal vene thrombosis, but not in patients with benign focal liver leasions. Determined in liver as a whole, HPI is a sensitive indicator of the presence of malignant liver tumours, but is within normal range in patients with hepatic hemangioma. The aim of the study was to investigate characteristic of blood flow in hemangioma itself, separately from but in relation to the liver blood flow, using hepatic radionuclide angiography. We have examined 12 patients with liver hemangioma confirmed mainly by positive 99mTc-labeled red blood cell scintigraphy, which diagnostic specificity for liver hemangiomas is near 100%. 8/12 hemangiomas resulted in photopenic areas on angioscintigrams, indicating lower blood flow, and rest were isoactive to surrounding liver tissue. Regions of interest have been delineated around the photopenic areas (hemangiomas) and surrounding liver tissue. Time-activity curves have been generated and slope of the fitted hepatic artery and (portal) venous portions of the hemangioma and liver curves have been determined. Perfusion indexes of hemangioma (PIH) and liver (HPI) have been calculated from the slopes, expressing portal venous flow as a portion of entire blood flow to the region. In addition, times of arrival and transit of intravenous bolus of 99mTc-pertechnetate through the hemangioma and liver tissue have also been derived from time-activity curves. Slope of the venous portion of the hemangioma time-activity curve is significantly lower then that of the venous portion of the liver curve (p < 0.01). So that, perfusion index of hemangioma (PIH = 0.34 +/- 0.12 (mean +/- SD) is significantly lower then hepatic perfusion index (HPI = 0.57 +/- 0.08) (p < 0.05). Bearing in mind interference of superimposed liver activity with that of hemangioma, these results indicate that liver hemangiomas are dominantly if not exclusively irrigated by hepatic artery branches. There are no data regarding relation between arterial and venous blood flow in liver hemangiomas determined by HRA. Obtained results are in harmony with arteriography data which confirm slow arterial blood flow through hemangiomas. Portal venous inflow of some angiomatous liver lesions in infants and children, and arterio-portal shunting in hemangiomas have been rarely reported. Results of this study indicate that regional determination of perfusion index and other HRA parameters in patients with focal liver lesion enables differentiation between tissues with different intensity and pattern of blood flow. The method could be used in examination of vascularisation pattern of other focal liver lesions. PMID- 11432250 TI - [The upper thoracic outlet vascular syndrome]. AB - A 16 patients with 20 vascular TOS have been evaluated at the our Institute. Fourteen of them were female, and 2 male patients, with average age of 33.1 (18 44) years. 19 of them had congenital, and one acquired TOS after trauma at neck shoulder region. 13 cases had arterial, and 7 venous TOS. In 10 cases a cause of TOS was cervical rib, in one scar tissue after clavicle fracture, while in 9 soft tissue anomalies. Eight cases with arterial TOS had a hand ischemia, one TIA and 5 periodical symptoms only during the arm hyperabduction. Two cases with venous TOS also had symptoms and signs during arm hyperabducrtion only, while five patients had axillary-subclavian deep venous thrombosis (DVT). All patients underwent CW-Doppler, Duplex-ultrasonographic and angiographic examination in normal position of the arm and during the hyperabduction. The four aneurysms of the subclavian artery, two poststenotic dilatation of the subclavian artery were found as well as one thrombosis of the axillary artery and 8 brachial artery embolism. The operative treatment consists from decompression and vascular procedure. A decompression procedure include 10 resections of the cervical rib, three transaxilary and 6 supraclavcular resection of the first rib, as well as one scalenectomy. A vascular procedures included 8 transbrachial thrombembolectomy and 4 resection and replacement of subclavian artery aneurysms. Four early complications were noticed: two partial pneumothorax, and two transiet medianus nerve paresis. The follow-up period was between one and six years (mean 3 years). In this period one (12.5%) late arterial occlusion was found. The vascular TOS is more rare than neurogenic, however in mostly cases requires surgical management. PMID- 11432251 TI - [Importance of adequate immobilization of the upper extremity in using the vascularized fibular graft]. AB - The most complications in patients after the reconstruction of the bone defect of the upper extremities are the fractures of the bone graft and pseudoarthrosis on one of the graft suture with defects of humerus. These complications influence on the primary efficiency of the reconstruction and prolong the recovery period. We registered five fractures, four pseudoarthrosis and two graft angulations in the prospective study on 25 patients where the defect of humerus, radius and ulna was compensated by the vascularized fibular graft. The analysis showed four fractures on the distal part of the reconstructed humerus graft and three pseudoarthrosis on the proximal part of reconstructed ulna. All osteogenic complications, apart from angulations which did not require additional interventions were solved by spongyoplasty according to Phemisters and more efficient postoperative immobilization. PMID- 11432252 TI - [Use of the internal thoracic artery in repeat myocardial revascularization surgery]. AB - The incidence of redo myocardial revascularization is increasing lately. These procedures are accompanied by the higher operative risk, and the use of internal thoracic artery graft may have additional negative impact on early-rization. Mortality and morbidity in this group did not differ significantly compared to a group where only venous grafts were used. Independent predictors of the unfavorable outcome (for both groups) were the presence of the chronic pulmonary disease, age over 65 and NYHA functional class IV. In the group with internal thoracic graft it was the age over 65, while in the group with only vein grafts it was chronic pulmonary disease. We have discussed potential problems during the redo procedure in patients in whom internal thoracic graft was used during the first procedure. PMID- 11432253 TI - [Total intravenous anesthesia using remifentanil and propofol with midazolam co induction in laparoscopic surgery of the gallbladder]. AB - We investigated effects of total intravenous anesthesia (TIVA) with propofol and remifentanil (in two parallel continuous infusions), on 28 ASA I-II patients undergoing laparoscopic cholecystectomy. All patients received midazolame (0.05 mg/kg b.w.), and 90 sec thereafter, remifentanil (0.5 g/kg b.w.). Computer controlled intravenous infusion of propofol started at dose of 6 mg/kg/h (by Graseby 3400 Syringe Pump). Muscle relaxation was achieved by rocuronium (0.6 mg/kg b.w.). After endotracheal intubation, rate of propofol was decreased on 3 mg/kg/h and started with another infusion of remifentanil (0.5 ug/kg/min). Before (T0) and after induction (T1), after start of surgery (T2), and at the end of surgery (T3), we evaluated: systolic, diastolic, and medial arterial blood pressure (SAP, DAP, MAP), heart rate (HR), peripheral saturation of O2 (O2Sat), and capnometry (ETCO2), by Datex-Engstrome AS/3 Monitore. It was followed side effects of anaesthesia, early and complete recovery rate, and frequency of nausea and vomiting in postoperative period. Results showed haemodynamic stability of patients after induction in anaesthesia (defined as decreasing of MAP 20%, compared with preinduction values). During investigation (T0-T3), results of 0.2Sat and ETCO2 were excellent (0.60 +/- 2 and 5.1 +/- 2.4 min). There wasonly one case of postoperative nausea and vomiting, and no significant side effects of anaesthesia. TIVA remifentanil-propofol and co-induction with midazolame makes possible haemodynamic stability of patients after induction in anaesthesia, good oxygenation during surgery, fast early and complete recovery, and avoiding of side effects of anaesthesia and postoperative nausea and vomiting. We concluded that it is a good choice of anaesthesia for laparoscopic cholecystectomy. PMID- 11432254 TI - [Comparative analysis in the use of natural and artificial vascular grafts on hematologic characteristics in patients]. AB - Investigation of two groups of patients was performed. Both male and female patients, 34 to 69 years old, in which extracorporeal circulation was established in the course of open heart and ascendental aorta surgeries were included in the investigation. The first group consisted of 20 patients with surgically revascularized miokard by triple venous grafts, and the second included 20 patients with partial replacement of ascendental aorta using synthetic dacron graft. It was concluded that extracorporeal circulation leads to a significant decrease of platelet count and agreability after surgery, as well as decrease of red blood cell count and hemoglobin and hematocrit value. However, contact of circulating blood with the synthetic dacron graft surface, in surgical procedures on ascendental aorta, leads consequently to a significantly higher decrease of platelet count and agreability, with the significant decrease of red blood cell count and decrease of hemoglobin and hematocrit value. Based on the obtained results it is possible to plan requirements for allogenous blood/red blood cells which are significantly higher in patients undergoing partial replacement of ascendental aorta using artificial dacron graft compared with patients after surgical revascularization of miocard with triple venous graft. PMID- 11432255 TI - [Assisted circulation. I. Systems with short-term mechanical circulatory support]. AB - The concept of artificial circulatory support has been established almost 200 years ago. It has only been within the last four decades that physicians and engineers have developed mechanical assist devices that can temporarily support the circulation until the native heart recovers from a reversible injury. If the heart does not recover sufficient function to maintain adequate hemodynamics, long-term circulatory support or permanent replace (biologically--heart transplant or permanent mechanical circulatory support) is needed. In this paper we describe the devices (intraaortic balloon pump, roller, centrifugal and axial pumps), that are in current clinical use for temporary (short-term) mechanical circulatory support. PMID- 11432256 TI - [The Mirizzi syndrome--from the first description until today]. AB - Pablo Luis Mirizzi was the first to describe an obstructive jaundice caused by compression of the common hepatic duct by the stone in the cystic duct or the neck of the gallbladder in 1948. McSherry et al in 1982. described a new type of Mirizzi's syndrome calling it type II. Csendes et al in 1989. gave a new classification in four types. According to it, type II of Mirizzi's syndrome was devided in three types depending on the size of the destruction of the common hepatic duct. We previously described a subtype of Mirizzi's syndrome in which besides very wide cholecystohepatic fistula, a combined fistula with duodenum was developed. Nagakawa et al in 1997. described a new type of Mirizzi's syndrom and gave their classification of syndrome in four types. In the present article the authors proposed a combined classification which takes into account not only all described variant of the syndrome but possibilities of operative reconstruction. Type I would be the same as in all previous classifications. Type II would correspond to the cholecystohepatic fistula in which a primary repair is possible. Type III would correspond to the cholecystohepatic fistula in which a primary repair is not possible so that biliodigestive anastomosis has to be carried out. Subtype IIIa would correspond to the same situations but complicated with fistula with the duodenum which has to be repaired as well. A Type IV of Mirizzi's syndrome would correspond to the inflammatory obstruction of the common hepatic duct as described by Nagakawa et al. PMID- 11432257 TI - [The development of anatomy]. AB - Doctors, particularly surgeons, realise the enormous importance of good knowledge of human anatomy today. It was not so in the past when doctors showed little or no interest for human anatomy for centuries. Dissections of the human body, necessary to study human anatomy, were either forbidden or limited to the corpses of criminals on whom capital punishment was carried out. The authors give a chronology of the development of human anatomy until 19. century when dissections of the human body became almost universally regulated with positive legislation. After the "golden age of surgery" began in 1870. surgeons gave an enormous contribution to anatomy. PMID- 11432258 TI - [Cytoreductive procedures in advanced primary ovarian carcinoma]. AB - Female patient, 68, hospitalized due to vaginal bleeding, anaemia and defecation disorder. We have done examinations as follows: clinical and laboratory exams, which confirmed severe anaemia. Explorative curretage of uterus--PH findings: malignant tissue. Rectoscopy: extraluminal compression at lo cm from the anal verge. Irigography: spasticity and extraluminal compression to proximal third of rectum and distal sigmoid colon. Ultrasound of abdomen: solid lobular formation 10 x 7 cm in diameter, on the left side of the urinary bladder, surrounded by thin layer of ascites. Ultrasound of liver: without signs of malignant disease. CT-scan of pelvis and abdomen: metastatic lymph nodes, up to 40 mm in diameter, alongside abdominal aorta; solid-lobular tumor, 10 x 7 cm, on the left side of urinary bladder, which fills the central portion of pelvic cavum. Secundary malignant deposits on the pelvic parietal peritoneum with minimal quantity of ascites (peritoneal carcinomatosis). Intraoperative findings confirmed malignant deposits on the right colon serosa, so we performed right hemicolectomy with ileo colo anastomosis, omentectomy, hysterectomy, bilateral adnexectomy, low anterior resection of the rectum, peritonectomy of pelvis and reconstruction of digestive tubus by colo-rectal anastomosis with circular stapler ILS 33. Duration of operation was 6:45 hours, blood loss was 1100 ml. Intraoperatively was intraperitonealy applicated Adriamycin. One month after the operation, systemic chemotherapy consisting of Endoxan and Karboblastin was administered, for the duration of 6 months, once monthly. Twelve months after the operation NMR showed normal findings, patient was without evidents of recurrent disease. PMID- 11432259 TI - [Mesothelial cyst and cystic mesothelioma of the greater omentum:case report and literature review]. AB - Mesothelial cysts and cystic mesotheliomas of the greater omentum are very rare clinical entities. About 30 cases of mesothelial cysts and only 4 cases of cystic mesotheliomas were described in the world literature so far. Over 13 years period (1987-2000), between 15,000 laparotomised adult patients, we have diagnosed one case of mesothelial cyst and one of cystic mesothelioma only. Mesothelial cyst, 7 cm in diameter, covered by regular mesothelial cells, was found accidently during gaster cancer surgery. In a 78 years old man, 30 cm in diameter cystic mesothelioma was diagnosed with immunohistochemical studies, necessarily for differentiation between cystic mesothelioma and other peritoneal cysts. Due to possible aggressive course at the diseases and tendency to recurrency, a complete surgical excision and log term follow-up are necessary. PMID- 11432260 TI - [Segmental portal hypertension as a rare cause of gastric hemorrhage--case report]. AB - Segmental portal hypertension is a rare pathologic condition, which produce gastric bleeding. Spleen vein thrombosis is more often caused by pancreatic disease (inflammations, tumors). Diagnosis is difficult to perform. Initial treatment is conservative. After successful conservative treatment early surgery should be planned. Unsuccessful conservative treatment indicates surgery. Splenectomy absolutely eliminates risk from rebleeding. Prognosis of these patients depends from etiology of pancreatic disease. We present a 46 old woman who successfully operated in our Department of Surgery due to massive upper gastrointestinal bleeding caused by isolated portal hypertension. PMID- 11432261 TI - [Creation of a new vagina using a part of the rectosigmoid colon]. AB - The paper shows the indications, preoperative and intraoperative evaluation as well as the way to make a new vagina. The authors advise to use part of colon sigmoideum to make a new vagina, this vagina is very much like the real one and the function is unlimited. During the operation the shortest possible part of colon sigmoideum should be taken in order to avoid oversecretion (mucorrhoea). After taking the segment and making termino-terminal anastomosis the tunnel should be made between the bladder and rectum and the segment should be stitched to the introitus of the vagina. For three weeks after the operation the patient should keep a tampon in the [figures: see text] new vagina and we suggest the dilatation of the introitus for three months. After the three months period we advise the sexual intercourse. The number of postoperative complications was minimal and the sexual intercourse was normal. PMID- 11432262 TI - Environmental monitoring in soil contamination and remediation programs: how practitioners are using the Internet to share knowledge. AB - Internet listservers provide a means for professionals from all sectors of the industry and profession, to communicate and collaborate with each other, as well as other stakeholders (e.g., suppliers, academics, the general public and community members) in real time. This article highlights key Internet listservers in the field of environmental monitoring in soil contamination and remediation and how to subscribe to them. The most active and relevant listservers for environmental scientists, technologists and professionals in the soil contamination and remediation profession are the Bioremediation Discussion Group (BioGroup), Phytonet, Phytoremediation listserver, Groundwater listserver and Environmental Forensics listserver. Other observations and lessons so far from the use of Internet listservers are: (i) that moderators provide an important role in maintaining the level of quality and participation, (ii) do not underestimate the knowledge base held within these, and (iii) if not selected and managed properly, e-mail from listservers can generate an excess of e-mail and waste time. PMID- 11432263 TI - Bioremediation of an area contaminated by a fuel spill. AB - In order to decontaminate a large area of restricted access contaminated by a fuel spill, laboratory and field studies were developed in two steps: (a) monitoring of the laboratory experiment on bacterial growth under aerobic and anaerobic conditions with and without addition of nutrients; and (b) use of the best conditions obtained in (a) for the decontamination of the soil. A hydraulic barrier was installed both to clean the aquifer and to avoid migration of hydrocarbons as a consequence of their solution in the groundwater and subsequent displacement. The objective was to create an ideal environment for the treatment of the affected area that favoured the growth of the indigenous bacteria (Pseudomonas and Arthrobacter) that biodegrade the hydrocarbons. Monitoring of the changes in the total concentration of petroleum hydrocarbons in the soil subjected to bacterial action was performed by gas chromatography. In a field study, the progress of biodegradation of hydrocarbons was evaluated in situ by changes in subsurface CO2/O2 levels by means of an analyser equipped with an infrared detector. Biostimulation and oxygen were the most influential factors for the biodegradation of the hydrocarbons. The use of bioventing of the soil was shown as an excellent technology to promote in situ bioremediation of the polluted area. PMID- 11432264 TI - Approaches to the remediation of a polychlorinated biphenyl (PCB) contaminated soil--a laboratory study. AB - A soil that had been historically contaminated with Aroclor 1242, 1248, 1254 and 1260 was decontaminated by two surfactant-mediated cleaning procedures that had been chosen to mimic ex-situ washing and in-situ soil flushing processes. A preliminary screening selected four surfactants (from 17 commercial formulations) for their ability to mobilise PCBs from the soil while suffering minimal losses to the supercritical carbon dioxide (scCO2) that was used in a separate back extraction procedure. The mobilisation was enhanced, with minimal foam formation, by the presence of 17% (v/v) IBMK in the surfactant suspension. Each of the four surfactants, at 1, 3, or 5% (v/v) concentration, was evaluated by (i) 15 successive 10 min sonication-filtrations and (ii) continuous soil column flushing during 20 h. Each filtrate from (i) and samples, taken at hourly intervals, from (ii) were analysed for their PCB and surfactant content. Both extraction procedures mobilised PCBs efficiently when extended for longer periods and were modelled accurately as the sum of a constant and single-term exponential increase to a maximum. The predicted number of replicate stages required to mobilise 50% of the toxicants (t50) varied from 7 to 3 for sonication-washing of the soil (10 g) or from 6.8 to 2.8 h for column flushing of 30 g soil and decreased as the concentration of surfactant in the aqueous phase was increased. The combined PCB laden aqueous suspensions were then back-extracted efficiently with scCO2 and the eluate was dechlorinated quantitatively as it traversed a short, heated column of silver-iron bimetallic mixture. PMID- 11432265 TI - Inter-organisational sampling trials for the uncertainty estimation of landfill gas measurements. AB - Inter-organisational sampling trials were performed at a reclaimed landfill site in Bedfont, West London, UK, to assess the uncertainty in measured concentrations of landfill gas. Eight participants independently measured concentrations of methane, carbon dioxide and oxygen, each on one occasion, over a two week period in February 1999. The organisers monitored the natural variability of the gas concentration over this sampling period using a control borehole in order to separate the variance that arose as a result of the sampling procedure from that caused by natural geochemical variation. A collaborative trial in sampling showed the extent of agreement between participants' results when they measured gases using the same, nominal protocol and equipment. The protocol was identified as fit for the purpose of estimating the mean concentration of methane, carbon dioxide and oxygen in a single borehole to within 21%, 15% and 321% of their assigned values, respectively (at 95% confidence). A realistic estimate of uncertainty in measurements of concentration from different protocols was estimated by the use of a sampling proficiency test, in which participants collected measurements using a protocol selected on the basis of their own professional judgement. Each participant's performance was compared with the consensus mean for each of the three gases using the z-score assessment proposed by the International Harmonised Protocol. This test highlighted the potentially large uncertainty in measured gas concentrations that can arise (91% relative uncertainty for CH4 in one borehole), and identified that an improved sampling protocol is required if misclassification of the explosive potential of methane is to be avoided. PMID- 11432266 TI - Variations in the estimation of the contribution of environmental tobacco smoke (ETS) to respirable (< or = 5 microns) indoor air particulates obtained by the use of different analytical methods. AB - Several methods are in use for the identification of the contribution of particulate associated environmental tobacco smoke (ETS) and sidestream smoke to an atmosphere. These include the measurement of respirable suspended particulates (RSP), measurements of the total UV absorption and total fluorescence emission of a methanol extract of collected particulates and the use of specific marker compounds such as solanesol and scopoletin. Use of these methods gave values for the contribution of particulate ETS to total respirable (< or = 5 microns) particulates in the ranges 8.3-124.7% for smokers' houses and 9.6-121.2% for smokers' offices, respectively. However, using what we consider to be the most reliable methodology, based on the measurement of solanesol, the average contribution of particulate ETS to total respirable (< or = 5 microns) particulates for smokers' houses was 21.7% and for smokers' offices was 23.3%. PMID- 11432267 TI - Exposure to airborne methacrylates and natural rubber latex allergens in dental clinics. AB - The exposure of dental personnel to airborne methacrylates and natural rubber latex (NRL) allergens was studied during placing of composite resin restorations in six dental clinics in Finland. Both area and personal sampling were performed, and special attention was paid to measurement of short-term emissions from the patient's mouth. Methacrylates were collected onto thermal desorption tubes filled with Tenax TA and NRL allergens onto membrane filters. The methacrylate samples were thermally desorbed and analysed by gas chromatography with mass selective detection. The NRL allergen concentrations were determined by the allergen-specific IgE-ELISA-inhibition method. The median concentration of 2 hydroxyethylmethacrylate (2-HEMA) was 0.004 mg m-3 close to the dental nurse's work-desk and 0.003 mg m-3 in the breathing zone of the nurse with a maximum concentration of 0.033 mg m-3. Above the patient's mouth the concentration of 2 HEMA was about 0.01 mg m-3 during both working stages, i.e., during application of adhesive and composite resins and during finishing and polishing of the fillings. Maximum concentrations of 3-5 times higher than median concentrations were also measured. Triethyleneglycol dimethacrylate was released into the air mainly during the removal of old composite resin restorations (0.05 mg m-3) and only to a minor extent during finishing and polishing procedures. The median concentration of the NRL allergen was 0.12 au m-3 (au = arbitrary unit) with a maximum concentration of 1.1 au m-3. The results show that, except for short-term emissions from the patient's mouth, the exposure of dental personnel to methacrylates and NRL allergens is very low. Measures to reduce exposure are discussed, as the airborne concentrations of methacrylates should be kept as low as possible in order to reduce the risk of hypersensitivity. PMID- 11432268 TI - Characterization of chemical interferences in the determination of unsaturated aldehydes using aromatic hydrazine reagents and liquid chromatography. AB - A systematic investigation on interferences in the determination of unsaturated aldehydes and ketones using the 2,4-dinitrophenylhydrazine (DNPH) method is described. Acrolein, crotonaldehyde, methacrolein and 1-buten-3-one are derivatized with DNPH in the presence of an acidic catalyst to form the respective hydrazones. The unstable hydrazones react with excess reagent to form adducts. These are identified by high-performance liquid chromatography (HPLC) mass spectrometry and spectroscopic techniques after cryogenic fraction collection of the adducts. The quantification of the unsaturated carbonyls with the DNPH method remains difficult. N-Methyl-4-hydrazino-7-nitrobenzofurazan (MNBDH) was used as an alternative reagent for this purpose. As with DNPH, the formation of a side product is observed. In contrast to DNPH, the alteration of the pH immediately after sampling leads to only one reaction product, which is stable and storable in solution at 4 degrees C for 2 days. PMID- 11432269 TI - HPLC combined with multiple mass spectrometry (MSn): an alternative for the structure elucidation of compounds and artefacts found in smog chamber samples. AB - Ion trap multiple mass spectrometry combined with high-performance liquid chromatography (online HPLC-MSn) was used to identify carbonyl compounds and artefacts found in air samples of 2-3 L taken during the OH-radical reaction study of cis-3-acetyl-2,2-dimethylcyclobutylethanal (pinonal) or cis-3-acetyl-2,2 dimethylcyclobutylcarbaldehyde (nor-pinonal) in a smog chamber. Carbonyls were transformed into 2,4-dinitrophenylhydrazones. Detection of low picogram quantities was possible. Twenty-eight compounds could be identified based on the presence of structure characteristic fragments, a developed fragmentation scheme, comparison of retention times and a library of MSn spectra of selected carbonyls. Furthermore, the additional elucidation of the sub-structures of sixteen unknowns was possible. Details about the structure elucidation procedure and the formation of the observed fragments are presented. PMID- 11432270 TI - Measuring the CO2 flux at the air/water interface in lakes using flow injection analysis. AB - The carbon dioxide flux at the air/water interface in lakes was calculated after the determination of H2CO3* (free CO2) and atmospheric CO2 using flow injection analysis (FIA) coupled to a conductometric detector. The method is based on the diffusion of CO2 through a hydrophobic membrane into a flow of deionized water, generating a gradient of conductivity proportional to the concentration of CO2 in the sample. Using one experimental set-up, the speciation of the inorganic carbon (H2CO3* and dissolved inorganic carbon) was accomplished by simply adjusting the sample pH. The determination of CO2 in the atmosphere was carried out by direct injection of the gaseous samples. The FIA apparatus was taken into the field and CO2 fluxes were evaluated in several Brazilian lakes. In these lakes, representing different eutrophic stages, the CO2 flux varied from -242 (invasive) up to 3227 (evasive) mumol CO2 m-2 h-1. PMID- 11432271 TI - Temporal variation of copper and zinc complexation capacity in the Humber estuary. AB - The complexation capacity (CC) for zinc and copper in the Humber estuary was determined and compared with data obtained using the same procedures from previous years. The CC for both metals was found to be relatively stable over a time span of 4 years for zinc and 10 years for copper. Levels of CC ranged from, in the upper estuary, 400 nM (copper) and 150 nM (zinc) to between 25 and 50 nM for both metals in the lower estuary. The CC from freshwater inputs appears to be relatively conservative with simple dilution accounting for the decrease in CC as salinity increases. A seasonal effect for copper is suggested. PMID- 11432272 TI - Radionuclides in raised bogs: a case study of Bor za Lasem. AB - Bor za Lasem is a raised bog in the Orawsko-Nowotarska Valley (southern Poland). About half of the Bor za Lasem area has been exploited while the rest is undisturbed. Peat samples from both parts of the bog were analysed. Measurements of 137Cs, 134Cs, 239,240Pu, 238Pu, 40K, 228Ra, and 226Ra activity concentrations in all samples were performed. Comparison of the results of radiocaesium and plutonium activity concentrations reveals the highest activity concentrations of radionuclides in the undisturbed part of the peat-bog area. The radiocaesium content decreased with depth, whereas the plutonium concentration had a maximum value at a depth of 15-20 cm. The lowest activity concentrations were found in the exploited part of the bog. Potassium and radium isotopes were detected only in shallow openings within the heavily exploited part of the bog, which might suggest partial mixing of the peat with the mineral sub-soil. The growth rate of the top layer of the undisturbed part of the bog obtained from plutonium distribution was estimated as approximately 0.5 cm per year. PMID- 11432273 TI - Representative sampling and sample preparation in biological environmental monitoring using spruce shoots. AB - The design of sampling and sample processing has a tremendous effect on the analytical results from which conclusions are drawn with respect to the quality of the environment and any possible impact on human health. Large scale environmental surveys need rigorous planning and extensive screening experiments to evaluate their boundary conditions. Representativeness and sample homogeneity are highlighted here in the context of a long-term biological monitoring survey for trace elements using spruce shoots. Several aspects concerning the selection of samples, amount of material, material processing and sample preparation prior to chemical characterisation are illustrated by selected examples. PMID- 11432274 TI - Sources of variability in collection and preparation of paint and lead-coating samples. AB - Chronic exposure of children to lead (Pb) can result in permanent physiological impairment. Since surfaces coated with lead-containing paints and varnishes are potential sources of exposure, it is extremely important that reliable methods for sampling and analysis be available. The sources of variability in the collection and preparation of samples were investigated to improve the performance and comparability of methods and to ensure that data generated will be adequate for its intended use. Paint samples of varying sizes (areas and masses) were collected at different locations across a variety of surfaces including metal, plaster, concrete, and wood. A variety of grinding techniques were compared. Manual mortar and pestle grinding for at least 1.5 min and mechanized grinding techniques were found to generate similar homogenous particle size distributions required for aliquots as small as 0.10 g. When 342 samples were evaluated for sample weight loss during mortar and pestle grinding, 4% had 20% or greater loss with a high of 41%. Homogenization and sub-sampling steps were found to be the principal sources of variability related to the size of the sample collected. Analysis of samples from different locations on apparently identical surfaces were found to vary by more than a factor of two both in Pb concentration (mg cm-2 or %) and areal coating density (g cm-2). Analyses of substrates were performed to determine the Pb remaining after coating removal. Levels as high as 1% Pb were found in some substrate samples, corresponding to more than 35 mg cm-2 Pb. In conclusion, these sources of variability must be considered in development and/or application of any sampling and analysis methodologies. PMID- 11432275 TI - Smart screening. PMID- 11432276 TI - Spring cleaning helps particulate levels. PMID- 11432277 TI - Asian haze could be a health risk. PMID- 11432278 TI - Chemicals policies become child-centred. PMID- 11432279 TI - Arsenic compounds may cause genetic damage. PMID- 11432281 TI - Managed care. Slow pay: the beast that won't go away. PMID- 11432280 TI - Pesticide information available from the World Health Organization. PMID- 11432282 TI - Information technology. Remote control. PMID- 11432283 TI - Payer relationships. How'm I doing? PMID- 11432284 TI - Red tape. The paperwork burden. PMID- 11432285 TI - Patient privacy. Press rips HIPAA. PMID- 11432286 TI - 1Q[3a]. Allow individuals to buy into the federal health plan? PMID- 11432287 TI - Design/construction. Score a TD for kids. PMID- 11432288 TI - Mission makes Wall Street. PMID- 11432290 TI - Upgrades climb. But slowly. PMID- 11432289 TI - Fixing finance. PMID- 11432292 TI - Inspiring ideas. PMID- 11432291 TI - Investment smarts. PMID- 11432293 TI - Demographic peristalsis. Implications of the age wave for gastroenterologists. AB - With the advent of the twenty-first century, a new phenomenon in human history emerged, one that is expected to dominate the social, economic, health, and health care agenda of the United States and the world. This so-called "age wave" represents an unprecedented demographic trend that will make the oldest cohorts, especially those older than 75, 85, and 100 years old, the fastest growing segment of the population for at least the next 50 years. PMID- 11432294 TI - Nutrition in the elderly. AB - Nutrition is a prime concern for the optimal health of older persons. Nutritional requirements for older persons must take into account the physiologic changes that occur with aging. The gastrointestinal tract is the site of key structural and functional changes that affect nutrient intake and assimilation. A working knowledge of nutritional screening, assessment, and interventions can assist the practitioner in providing quality care for the older patient. PMID- 11432295 TI - Esophageal disorders in the elderly. AB - As the number of elderly in the population increases, decompensation of swallowing and airway protective mechanisms can create an increased health care burden. This article outlines the effect of aging on deglutitive function and esophageal and aerodigestive reflexes. Specific disorders in the elderly are discussed. PMID- 11432296 TI - Acid peptic disease in the elderly. AB - GERD and peptic ulcer disease are important diseases in the elderly. GERD presents similarly in the elderly and the young, although elderly patients may have less severe symptoms yet more severe mucosal disease and a higher prevalence of BE. Although the prevalence of H. pylori is falling, the elderly remain at risk for peptic ulcer because of the widespread use of NSAIDS. The presentation of peptic ulcer disease in the elderly can be subtle and atypical when compared with younger patients, leading to a delay in diagnosis. Because of comorbidity in the aged, peptic ulcer disease and its complications result in increased morbidity and mortality rates. PMID- 11432297 TI - Gastrointestinal bleeding in the elderly. AB - Among patients with acute gastrointestinal bleeding, older age is associated with an increased rate of comorbidity, greater medication use, and atypical clinical presentations. The aging of the population makes the evaluation and management of gastrointestinal bleeding in the elderly a special and increasingly common clinical challenge. The unique features and common causes of upper and lower gastrointestinal bleeding in the elderly are reviewed. Important management issues considered include hemodynamic resuscitation; anticoagulation; and medical, surgical, and endoscopic therapy. PMID- 11432298 TI - Inflammatory bowel disease in the elderly. AB - Approximately 15% of all patients with IBD first develop symptoms after age 65. As the number of elderly in the population continues to grow, clinicians should expect to see a greater number of elderly IBD patients. In general, the presenting features of IBD are similar to those encountered in younger patients, but the broad differential diagnosis of colitis in the elderly can make definitive diagnosis more challenging. Although most therapies for IBD have not been studied specifically in the elderly, as a general rule, medical and surgical treatment options are the same regardless of age. Osteoporosis, a condition generally associated with aging, should be managed aggressively in patients with IBD because many older persons already have a substantial baseline risk for accelerated bone loss. PMID- 11432299 TI - Diarrhea and malabsorption in the elderly. AB - Physicians who care for elderly patients should be alert to the possible presence of diarrhea and malabsorption. Older patients may not admit to having chronic diarrhea, particularly if they also are incontinent. If diarrhea is of short duration, an infectious cause is at least as common as in the young. Institutionalized elderly are particularly prone to gastrointestinal infections, but the manifestations may not be overt. When an intestinal infection and potential medication-induced gastrointestinal disturbances have been excluded, the differential diagnosis of diarrhea in the elderly is the same as in the young. Causes include intestinal malabsorption, even though diarrhea is a less common manifestation of malabsorption in the old than in younger patients. In the elderly, micronutrient deficiency is a common presenting clinical picture; because the symptoms of malabsorption are covert, the diagnosis often is delayed, and nutritional deficiencies are more common and more severe than in the young. Because the elderly have less nutritional reserve than the young, these deficiencies are clinically much more devastating in the elderly. Although the causes of malabsorption, as a whole, are similar in older and younger patients, chronic pancreatic insufficiency of unknown cause and intestinal bacterial overgrowth without an anatomic abnormality of the small intestine are syndromes that are specific to the elderly and must be considered in any older patient with unexplained weight loss or failure to thrive. Often, therapeutic trials are necessary to establish a potential diagnosis. PMID- 11432300 TI - Ischemic bowel disease in the elderly. AB - The ischemic bowel diseases are a heterogeneous group of disorders usually seen in elderly individuals. They represent ischemic damage to different portions [figure: see text] of the bowel and produce a variety of clinical syndromes and outcomes. Colonic ischemia is the commonest of these disorders and has a favorable prognosis in most cases. In contrast, acute mesenteric ischemia, most commonly caused by a superior mesenteric artery embolus, is a disease with a poor prognosis. Acute mesenteric ischemia secondary to nonocclusive mesenteric ischemia usually is a [figure: see text] catastrophic complication of other severe medical illnesses, most notably atherosclerosis. Proper diagnosis and management of patients with ischemic bowel disease requires vigilance on the part of the physician and a willingness to embark on an aggressive plan of diagnosis and management in the appropriate setting. PMID- 11432301 TI - Diverticular disease in the elderly. AB - Diverticular disease is common among the elderly. Because of the advanced age and muted symptoms and signs of many of those affected, diagnosis can be difficult. Consequently, great demands are placed on the physician to diagnose and treat clinically evident diverticular disease. Endoscopic, radiologic, and surgical advances have increased the availability of more definitive therapies for patients with complicated diverticular disease and diverticular hemorrhage. PMID- 11432302 TI - Constipation and fecal incontinence in the elderly. AB - Although constipation and fecal incontinence are common symptoms in the elderly, relatively little research has been done to differentiate physiologic changes in rectoanal function resulting from aging and pathologic changes resulting from diseases occurring as patients age. Certain physiologic changes occur in many older patients and may predispose them to the development of constipation or fecal incontinence. These symptoms need the same thoughtful evaluation and management in the elderly as in younger patients. Results of therapy often can be good, leading to alleviation of suffering and the ability to lead a fuller life. PMID- 11432303 TI - Functional abdominal pain in the elderly. AB - Although aging per se affects function throughout the gut, particularly after age 70, the observed changes are relatively modest and often asymptomatic, perhaps because of the vast reserve of neuromuscular functional elements in the gut. The proximal esophagus, anus, and pelvic floor are possible exceptions to this generalization, and the combination of aging and factors such as minor strokes or obstetric damage often results in dysphagia, constipation, or fecal incontinence. Managing elderly patients with functional abdominal pain demands clinical acumen, tact, understanding, and patience. Further studies are required to elucidate the consequences of aging on gastrointestinal sensorimotor function at several levels of the digestive tract. With the expected explosion in the proportion of the population older than age 75, this field should become a high priority for clinical and research efforts. Also required will be a major campaign to educate patients and practitioners and multidisciplinary collaborations among primary care practitioners, gastroenterologists, psychologists, and physiatrists to provide optimal pain management in the elderly. PMID- 11432304 TI - Pancreatic and biliary disorders in the elderly. AB - Structural and functional changes in the biliary tract and pancreas associated with advanced age are well documented in the literature and must be taken into account in evaluating patients with possible biliary and pancreatic disorders. The relationship between normal, age-related physiologic changes and various pancreatico-biliary diseases is not well defined. Elderly patients may present with severe biliary and pancreatic disease that may pose difficult management problems because of coexisting medical illnesses. Despite these challenges, all but the most frail elderly patients can benefit from appropriate medical, endoscopic, and surgical therapy. PMID- 11432305 TI - Liver disease in the elderly. AB - Despite several morphologic and functional changes that have been described in the aging liver, most relevant studies fail to identify a significant age-related deficit in liver function in humans. One of the important age-related changes is a decrease in regenerative capacity, which may partly explain the impaired recovery after severe viral and toxic injury in the elderly. Nevertheless, livers from older subjects are used successfully for transplantation. Substantial morbidity and mortality in the elderly is attributable to liver diseases, and the number of patients older than 65 years of age with chronic liver disease is increasing rapidly. Although there are no liver diseases specific to advanced age, the presentation, clinical course and management of liver diseases in the elderly may differ in important respects from those of younger individuals. PMID- 11432306 TI - Gastrointestinal tract cancer in the elderly. AB - Gastrointestinal cancers are common in the elderly. In the 1990s, modest advances in the early diagnosis, staging, and treatment of gastrointestinal cancers were made. Emphasis has been placed on screening for colorectal cancer; development of new diagnostic and staging modalities, particularly endoscopic ultrasound; and introduction of new treatment regimens, such as chemoradiation programs for esophageal, pancreatic, and colorectal cancers as well as endoscopic mucosal resection of early gastrointestinal cancers. New palliative techniques, such as laser and photodynamic therapy and placement of enteral stents, are being used increasingly in patients who have advanced cancer and are not surgical candidates. In the past, attitudes toward the elderly affected the management of cancer. Age should not be the only parameter considered when addressing the treatment of a gastrointestinal malignancy. Management decisions in the elderly should follow the same principles as those in younger patients. A thorough medical evaluation in the elderly is necessary to evaluate the patient's risk and to optimize surgical, chemotherapeutic, and palliative outcomes. PMID- 11432307 TI - Risk factors for mortality in primary isolated coronary artery bypass grafting surgery. AB - BACKGROUND AND PURPOSE: Identifying the risk factors for mortality in coronary artery bypass grafting (CABG) surgery is important to improve surgical results. The purpose of this study was to identify the risk factors for mortality in primary isolated CABG in a series of Taiwanese patients. METHODS: Medical records of 914 patients who underwent primary isolated CABG surgery in Veterans General Hospital-Taipei during the period from January 1, 1991, to December 31, 1995, were reviewed. Eighteen clinical and seven operative variables were included in the univariate and multivariate analyses to identify the determinants of mortality in CABG surgery. RESULTS: Thirty-one patients (3.4%) died within 30 days after surgery; 41 (4.5%) died during hospitalization for the procedure. After univariate and multivariate analyses, emergency surgery, history of myocardial infarction, concomitant peripheral artery occlusive disease (PAOD), and prolonged cardiopulmonary bypass (CPB) were found to be significant determinants of mortality in primary isolated CABG. Other variables, including age, sex, angina class, NYHA class, diabetes mellitus, the number of anastomoses, aortic cross-clamp time, stenosis of the left main coronary artery, the number of stenotic coronary arteries, history of congestive heart failure, and a left ventricular ejection fraction less than 35%, were not significant determinants of mortality. Patients undergoing emergency surgery were found to be at highest risk of mortality. CONCLUSIONS: Patients undergoing emergency surgery who had a history of myocardial infarction, concomitant PAOD, or prolonged CPB were at higher risk of mortality in CABG surgery. More comprehensive techniques in myocardial protection, surgical procedures, and postoperative care should be used in the treatment of high-risk patients to reduce mortality. PMID- 11432308 TI - Relationship of duodenal ulcer recurrence to gastric metaplasia of the duodenal mucosa and duodenal bulb deformity. AB - BACKGROUND AND PURPOSE: This study investigated changes in the severity of gastric metaplasia (GM) of the duodenal mucosa before and after ulcer healing and Helicobacter pylori eradication. It also investigated whether deformity of the duodenal bulb affects the severity of GM and the likelihood of ulcer recurrence. METHODS: Eleven patients were consecutively enrolled in this study. They all had duodenal ulcer(s) and H. pylori infection, for which they had received anti-H. pylori triple therapy during the active ulcer stage, and had all undergone serial endoscopic examinations during both the active ulcer and scarring ulcer stages, and at 1 year after ulcer healing. Duodenal biopsies were obtained at each endoscopy to identify the severity of GM. Duodenal ulcers were divided into three types by bulbar shape and GM was classified into four grades of severity. RESULTS: All 11 patients had increased GM severity just after ulcer healing. The 1-year follow-up study revealed that the GM was unchanged in six of eight patients with grade 3 GM severity at the scarring stage, while in the other two it regressed to grade 1 or 2; these two patients suffered ulcer recurrence. A markedly deformed bulb (type III) was found in three patients, of whom two had ulcer recurrence. CONCLUSION: Two characteristic conditions were found in patients with duodenal ulcer recurrence after H. pylori eradication: a markedly deformed bulb with grade 3 GM at the scarring stage, and a change in GM from high to low grade at or around the previous ulcer site after ulcer healing. PMID- 11432309 TI - Urodynamic results of intravesical heparin therapy for women with frequency urgency syndrome and interstitial cystitis. AB - BACKGROUND AND PURPOSE: Although intravesical heparin instillation is effective in relieving symptoms in patients with frequency urgency syndrome and interstitial cystitis (IC), its efficacy has not been evaluated by urodynamic study. We used urodynamic studies to evaluate the efficacy of heparin instillation in patients with frequency urgency syndrome and IC. METHODS: Forty women (mean age, 59.6 yr) with severe frequency, urgency, and suprapubic pain at full bladder were enrolled in this study. Patients underwent videourodynamic study with the potassium chloride (KCl) test. All patients had a positive KCl test and were treated with intravesical heparin 25,000 units twice a week for 3 months. Results of urodynamic study and voiding symptom changes assessed by the International Prostate Symptom Score at the end of treatment were compared with baseline data. RESULTS: Twenty-nine patients had symptom score improvement of more than 50%, and eight had symptom score improvement of less than 50% but improved nocturia. Significant improvement in symptom score (9.0 +/- 4.0 vs 19.5 +/- 4.6, p < 0.001) and nocturia (2.3 +/- 1.1 vs 5.7 +/- 2.0, p < 0.001) were noted after treatment. Urodynamic study at the end of treatment revealed significant improvements in the first sensation of filling (146 +/- 55.4 vs 96 +/ 46.4 mL, p = 0.001) and cystometric capacity (304 +/- 84.8 vs 262 +/- 89.8 mL, p = 0.002). Posttreatment KCl test was negative in 20 patients, improved in 13, and unchanged in seven. Among the 10 patients with cystoscopically proven IC, eight had symptomatic improvement and four had a negative KCl test after treatment. CONCLUSIONS: IC and frequency urgency syndrome may be caused by increased urothelial permeability. The results of this study show that intravesical heparin can relieve bladder symptoms in a significant proportion of patients, and this may be associated with the restoration of mucosal integrity. PMID- 11432310 TI - Ultrasound examination of patellar tendon after harvest for anterior cruciate ligament reconstruction. AB - BACKGROUND AND PURPOSE: Reconstruction of ruptured anterior cruciate ligament (ACL) using the middle-third of the patellar tendon has become an established procedure. However, current studies of donor site morbidity have reported discrepant results. This study used ultrasound imaging to evaluate morphologic changes in the patellar tendon after harvest for ACL reconstruction. METHODS: Twenty-nine patients who had undergone successful ACL reconstruction with patellar tendon graft more than 1 year previously were enrolled in the study. Ultrasound imaging was used to compare the width, thickness, and tissue reaction of patellar tendons between the operated and contralateral sides. RESULTS: The operated side was 4.8% wider on average than the contralateral side (p < 0.05). The operated side was also 58.3% thicker on average (p < 0.0001), with irregular margins and heterogenous echogenecity. CONCLUSION: The donor site of the patellar tendon was significantly wider and thicker and was heterogenous when examined on sonogram more than 1 year postoperatively. PMID- 11432311 TI - Combined measurement of fetal nuchal translucency, maternal serum free beta-hCG, and pregnancy-associated plasma protein A for first-trimester Down's syndrome screening. AB - PURPOSE: It has been proposed that first-trimester Down's syndrome screening has a higher detection rate compared to second-trimester biochemical screening. This study investigated the accuracy of Down's syndrome screening during gestational weeks 10 to 13 using the combination of fetal nuchal translucency (NT) measurement with maternal serum concentrations of free beta-human chorionic gonadotropin (beta-hCG) and pregnancy-associated plasma protein-A (PAPP-A). METHODS: A total of 1,514 women with singleton pregnancies were enrolled in this study. Fetal NT was measured using the criteria published by the Fetal Medicine Foundation. Maternal serum concentrations of free beta-hCG and PAPP-A were determined by microtiter-plate ELISA. Down's syndrome risk was calculated using multivariate Gaussian distribution and Alpha software. RESULTS: Seventeen (1.12%) of the 1514 screened pregnancies had a fetal NT of at least 3 mm, and 41.2% of these had a poor pregnancy outcome, including four fetal aneuploidies. The odds of a fetal aneuploidy when the NT was greater than 2.0 multiples of median (MoM) was 90, when serum PAPP-A concentration was less than 0.45 MoM, it was 8.6, and when serum free beta-hCG concentration was greater than 2.2 MoM, it was 4.7. Using a risk cut-off level of 1 in 400, nine of 10 fetal aneuploidies were identified with a 4.7% false-positive rate, including two with trisomy 21, one with trisomy 18, and three with Turner's syndrome. CONCLUSIONS: This study demonstrated that Down's syndrome screening using the combined test in the first trimester had a higher detection rate than that of serum screening in the second trimester. Implementation of NT measurement in the first trimester provides substantial advantages for Down's syndrome detection and early diagnosis of fetal structural abnormalities. PMID- 11432312 TI - Effects of zinc deficiency on the vallate papillae and taste buds in rats. AB - BACKGROUND AND PURPOSE: Zinc deficiency is associated with multiple clinical complications, including taste disturbance, anorexia, growth retardation, skin changes, and hypogonadism. We investigated the zinc-deficiency-induced morphologic changes in the vallate taste buds of weanling and young adult male Wistar rats. METHODS: A total of 24 weanling and 30 young adult rats were used. Each age group was further divided into a control group fed a zinc-adequate (50 ppm) diet, a zinc-deficient (< 1 ppm) diet group, and a zinc-adequate pair-fed group who were fed the same amount of food as that taken by the zinc-deficient group. Weanling rats were fed for 4 weeks and young adult rats were fed for 6 weeks. The morphometry and morphologic changes of vallate taste buds were analyzed using light and transmission electron microscopy. RESULTS: Light microscopy revealed no significant difference in papilla size and morphology among the various groups. In both weanling and young adult rats in the zinc deficient diet and pair-fed groups, the number of taste buds per papilla (per animal) and the average profile area of the taste bud were significantly smaller than those of the corresponding controls (p < 0.05). Ultrastructural changes were seen only in the taste buds of weanling rats fed the zinc-deficient diet, with derangement of the architecture of the taste bud and widening of the intercellular space between taste bud cells. The proportion of type I taste bud cells in the taste buds of weanling rats fed the zinc-deficient diet decreased from 59% to 39%, and that of type II taste bud cells decreased from 25% to 12%. No obvious changes in the ultrastructure of type III taste bud cells were observed. CONCLUSIONS: The main effects of zinc deficiency in weanling and young adult rats and in adequate diet pair-fed rats were changes in the number and size of taste buds, and fine structure changes in the taste bud cells, especially during the accelerated growth stage after weaning. PMID- 11432313 TI - Coexistence of tuberculous constrictive pericarditis and right atrial tuberculoma: a case report. AB - Tuberculous constrictive pericarditis is a rare condition with a high mortality rate. The coexistence of constrictive pericarditis and intracardiac tuberculoma has not previously been reported. We report the case of a 65-year-old man presenting with left-side pleural effusion and signs of systemic venous congestion for 2 months. Echocardiography and computerized tomography showed a thickened pericardium and a mass in the right atrium. Pericardiectomy and excision of the right atrial mass were performed. Pathologic examination of the pericardium and the right atrial mass both revealed chronic granulomatous inflammation with acid-fast bacilli and confirmed the diagnosis of tuberculous constrictive pericarditis and right atrial tuberculoma. This case reminds us of the possibility of this type of rare combination of tuberculous constrictive pericarditis and intracardiac right atrial tuberculoma, and the need for complete imaging studies when such cases are encountered. PMID- 11432314 TI - Intractable ventricular tachycardia in a patient with left ventricular epicardial lipoma. AB - Primary cardiac lipomas are rare and are often reported as incidental findings during operations or autopsies. Lipoma-induced arrhythmia, especially ventricular tachycardia (VT), is an unusual complication of primary cardiac lipoma. We describe the case of a 43-year-old woman who presented with VT without any cardiac abnormality except for a left ventricular tumor. Electrophysiologic study (EPS) failed to detect the origin of the arrhythmia, although VT could be induced by EPS preoperatively. Due to suspicion of a relationship between the tumor and the VT, wide tumor resection was performed using cardiopulmonary bypass support. The excised lipoma measured about 10 x 8 x 3 cm and weighed 120 g, and was without pathohistologic evidence of malignant change. The patient was well without further postoperative need for antiarrhythmic drugs at 36 months' follow up, and VT was no longer inducible on subsequent EPS mappings. PMID- 11432315 TI - Life-threatening refeeding syndrome in a severely malnourished anorexia nervosa patient. AB - Overzealous refeeding in chronically malnourished anorexia nervosa patients may cause life-threatening complications. We describe a 14-year-old girl with anorexia nervosa who had a decrease in body weight from 45 kg to 25.5 kg over an 18-month period. She received 40 kcal.kg-1.d-1 carbohydrate-rich nutrition via enteral and parenteral routes. Her serum phosphate concentration dropped from a baseline of 1.39 mmol/L (4.3 mg/dL) to 0.19 mmol/L (0.6 mg/dL) on Day 4 of refeeding. Concurrent with the development of hypophosphatemia, she became drowsy and developed generalized muscle weakness, impaired myocardial contractility, thrombocytopenia, and gastrointestinal bleeding. Fluid overload with pulmonary edema complicated her recovery from these adverse events. After intravenous phosphate supplementation and fluid restriction, the symptoms of refeeding syndrome gradually resolved within 2 weeks. In chronically malnourished anorexia nervosa patients, nutritional support should be instituted gradually to avoid rapid electrolyte shifts and fluid overload. Serum phosphate concentrations, fluid status, and blood cell counts should be closely monitored. PMID- 11432316 TI - In utero progressive pulmonary stenosis successfully treated with transcatheter intervention after delivery. AB - It is unclear whether pulmonary stenosis with intact ventricular septum is a secondary cardiac malformation. We report an infant with pulmonary stenosis (diagnosed by fetal echocardiography) with progressive obstruction in late gestation who presented with increasing transvalvular pressure gradients (15 mm Hg at 22 weeks' gestation to 47 mm Hg at 35 weeks). The tricuspid/mitral valve annulus ratio decreased from 1.25 at 24 weeks' gestation to 0.96 at 33 weeks. At 38 weeks' gestation, a male infant weighing 3,524 g, with Apgar scores of 9 and 9 at 1 and 5 minutes, respectively, was delivered by cesarean section. Critical pulmonary stenosis was confirmed by postnatal catheterization. These findings support the postulation that pulmonary stenosis is a progressive disorder. After percutaneous balloon dilatation, the transvalvular pressure gradient decreased and the right ventricular cavity increased gradually. The transvalvular pressure gradient had decreased to 15 mm Hg and the tricuspid/mitral valve annulus ratio was 0.93 at the age of 2 years. PMID- 11432317 TI - Transmission of human immunodeficiency virus through bone transplantation: a case report. AB - Human immunodeficiency virus (HIV) can be transmitted through tissue or organ transplantation. Since 1988, eight cases of bone transplantation-associated HIV infection have been reported. We describe a 34-year-old woman with osteoarthritis, who was hospitalized in 1996 because of painful locomotion and deformity in the right knee. An old fracture of the femur and genu varum in the right knee was repaired by open reduction and internal fixation, with allogeneic bone transplantation for defect reconstruction. Five months later, she presented at another hospital with pain and swelling of the right thigh, where screening for HIV was positive. The transmission was tracked to a 50-year-old man with no history of HIV screening, from whom the femoral head was procured when he underwent a hip replacement for left femoral neck fracture. This is the first reported case of HIV infection through bone transplantation in Taiwan, and the preventable nature of this transmission should be underscored. PMID- 11432318 TI - K-ras mutation at codon 12 in stage I pancreatic adenocarcinoma: analysis by laser capture microdissection and direct sequencing. AB - Pancreatic ductal adenocarcinoma has been reported to carry a rate mutation high in codon 12 of the K-ras oncogene. To avoid the pitfalls of conventional methods of tissue dissection that might affect the sensitivity and specificity of detecting K-ras mutation, laser capture microdissection (LCM) technique was used. Pancreatic adenocarcinoma tissues were obtained from 15 patients who underwent Whipple's procedure. Selected tissues procured by LCM were analyzed by direct sequencing after polymerase chain reaction amplification of K-ras sequences at codon 12. K-ras mutation was noted in nine patients. All mutations showed G to A substitution at codon 12. The mutational pattern (GGT to GAT) is similar in both western and eastern reports. LCM is a feasible method to effectively obtain pure tumor cells from a surgical specimen. It remains to be determined whether this low mutation rate is a result of relatively early stage of disease or different carcinogenesis in different geographic regions. PMID- 11432319 TI - [Occurrence of the metabolic syndrome in the population of the town of Novi Sad]. AB - INTRODUCTION: Inadequate nutrition and physical inactivity are dominant health problems in general population resulting in high incidence and prevalence of noncommunicable diseases. Obesity (especially with central fat distribution), insulin resistance, hyperinsulinemia, hyperlipoproteinemia, dyslipoproteinemia and cardiovascular diseases are the main components of metabolic syndrome- Syndrome X. MATERIAL AND METHODS: Frequency of metabolic disorders characteristic for syndrome X are determined in a representative sample of Novi Sad town (1600 persons of both sexes, aged 25-64, of all professional levels). The response was 79.37%, so the sample included 1270 persons, in whom the following examinations were performed: history data, anthropometric measurements (height, weight, body mass index, weist to hips ratio), physical examination, blood pressure, biochemical analysis (fasting glycemia, oral glucose tolerance test (OGTT), total serum cholesterol, HDL-cholesterol, LDL-cholesterol, triglycerides). THE AIM: The aim of this research was to establish the frequency of obesity, types of obesity, glycemic status, disorders of glucose metabolism, disorders of lipid metabolism, frequency and type of hypertension, measurement of insulinemia and finally frequency of metabolic syndrome. RESULTS: The sample included 1270 persons. Among them 45.41% males and 32.64% females are obese. Extremely obese are 19.70% males and 27.12% females. W/H index > 1.0 was determined in 9.18% males while 48.82% females have W/H index 0.8-1.0. Fasting hyperglycaemia is present in 30.22% males and 20.86% females. Serum cholesterol > 5.20 mmol/l is determined in 74.96% males and 72.24% females. Arterial hypertension is diagnosed in 29.88% males and 23.40% females. In 82.75% persons of both sexes, aged 25-64, pathologic values among some of the followed-up characteristics are detected. In our sample syndrome X was suspected in 13.62% persons of both sexes (probability 50%) while 86.38% persons of both sexes are without risk for syndrome X (probability 70%). The best characteristic for dividing these two groups is W/H index (discriminatory coefficient 0.39). DISCUSSION AND CONCLUSION: In obese persons, especially in central obese persons, an active search for metabolic disorders which are present in syndrome X should be performed, because in that case measures of primary prevention can be applied. PMID- 11432320 TI - [Drug use during pregnancy, labor and the puerperium and after the Drug Use in Pregnancy Study]. AB - INTRODUCTION: An international study on drug use in pregnancy (DUP) started in 1987. There were 22 countries involved, among which two centers from Yugoslavia: Novi Sad and Zagreb. The investigation in Novi Sad comprised 296 parturients. On the basis of obtained results, education of physicians-gynecologists has been organized. 10 years later, a check-up study of 100 parturients was introduced in order to find out possible changes in the applied pharmacotherapy. MATERIAL AND METHODS: An investigation on drug use in pregnancy, delivery and puerperium has been performed at the Department of Obstetrics and Gynecology in Novi Sad. It included 296 parturients in 1989 (1st investigated period) and 100 parturients in 1999 (2nd investigated period). Results obtained in 1999 were compared with the results from 1989. RESULTS AND DISCUSSION: Chronic use of drugs was found similar for both investigated periods (2.38% in 1989 vs 5% in 1999). In this group there were no drugs which could endanger the fetus. In 1989, there were 12.6% of pregnant women using preparations such as iron supply, versus 54% in 1999, so that trends in prevention of pharmacotherapy of anemia approached those in developed countries. In the second investigated period there was a significant decrease in progresterone therapy and two times increased use of beta-adrenergic agonists, which is in accordance with new trends in pharmacology. Use of paracetamol decreased, whereas use of metamizole, known by serious side-effects, increased. Drug administration during delivery decreased in 1999 (77%) in relation to 1989 (94.93%) mostly comprising drugs for autonomous nervous system, analgesics and analgoantipyretics. Administration of ergot-alkaloids decreased: from 65.8% in 1989 to 22% in 1999. Use of drugs after delivery can endanger both the parturient and lactation, i.e. nutrition of the newborn infant and consequently the newborn itself. During 1989 practically every parturient (95.27%) was administered drugs, out of which about 97% belonged to the group of ergot-alkaloids which were routinly administered for acceleration of uterine involution. In 1999 ergot-alkaloids were administered only in 9% of delivered women. Use of oxytocins was also reduced because they were administered not routinely but only on purpose. CONCLUSIONS: The following conclusions were made on the basis of obtained data: 1. During the first period there was a difference in drug use when compared to the countries with developed pharmacotherapeutic practice. 2. Use of progesterones was remarkably decreased in 1999 versus two times increased use of beta-adrenergic agonists administered in prevention of spontaneous abortions. 3. Use of opiate analgesics during delivery was significantly reduced from 43% to 11% in 1999. 4. In the second investigated period use of iron preparations was significantly increased versus decreased use of ergot-alkaloids. 5. Use of paracetamol decreased, whereas use of metamizole (known by serious side-effects) increased. PMID- 11432321 TI - [Classification of dysphonias based on the primary etiologic factor (part I)]. AB - INTRODUCTION: Phonation is a complex integral function of the organism. Regular phonation is characterized by: clarity and adequate pitch. Dysphonia is a disorder of phonation. It may have many acoustic forms, but hoarseness is the best known symptom of dysphonia. Acoustic phenomena are caused by: aperiodicity of vocal vibration, turbulent air flow in the glottis and incomplete glottis closure. PREVIOUS CLASSIFICATIONS OF DYSPHONIAS: The best known classification of dysphonias was introduced by Perello. There are two groups: organic dysphonias and functional dysphonias. On the 8th Congress of Union of European Phoniatricians, in Koszeg (Hungary, 1979), Majdevac proposed a new classification. CLASSIFICATION OF DYSPHONIAS: We are proposing a new classification, made according to the primary etiologic factor in dysphonias. In this paper, we shall consider the first four. I DYSPHONIAS CAUSED BY PRIMARY FUNCTIONAL DISORDERS: This group includes: 1. Hyperkinetic dysphonia grade I 2. Hyperkinetic dysphonia grade II 3. Hypokinetic dysphonia 4. Contact hyperplastic dysphonia 5. Dysodic dysphonia II DYSPHONIAS CAUSED BY PRIMARY NEUROGENIC DISORDERS: This group includes: 1. Central dysphonias 2. Spasmodic (spastic) dysphonia 3. Dysphonia caused by myasthenia gravis 4. Dysphonia within skull base syndromes 5. Dysphonia caused by unilateral palsy of the inferior laryngeal nerve 6. Dysphonia caused by bilateral palsy of the inferior laryngeal nerve 7. Dysphonia caused by palsy of the superior laryngeal nerve III DYSPHONIAS CAUSED BY PRIMARY PSYCHOGENIC DISORDERS: This group includes: 1. Psychogenic aphonia 2. Psychogenic dysphonia 3. False mutation IV DYSPHONIAS CAUSED BY PRIMARY SOMATIC DISORDERS: This group includes: 1. Dysphonia caused by insufficiency of vocal cords 2. Dysphonia caused by oedema of vocal cords 3. Dysphonia caused by laryngitis (secondary functional) 4. Cord-ventricular voice 5. Posttraumatic dysphonia 6. Arthrogenic dysphonia 7. Presbyphonia CONCLUSION: Dysphonia is a disorder of phonation which originates at the glottis level. When disorders of phonation are concerned it is necessary to study the organism as a whole as well as all mechanisms which take part in voice production. In that case the damaged part of the phonation system can be diagnosed, which enables efficient medical treatment of the disorder. PMID- 11432322 TI - [Etiopathogenesis, clinical picture and diagnosis of onychomycoses]. AB - ETIOLOGY OF ONYCHOMYCOSES: Onychomycoses can be caused by dermatophytes, molds and yeasts. However, dermatophytes appear to be the chief organisms capable of a primary attack on the nail. By far the most frequent dermatophytes isolated from nails are Trichophyton rubrum, T. mentagrophytes var. interdigitale and Epidermophyton floccosum. Molds virtually only invade toenails, but their role as a primary pathogen is still debated. Yeasts have been isolated from diseased nails at highly different rates. Nails may be infected by two different dermatophytes, two dermatophytes and a yeast, a dermatophyte, a yeast and a mold, etc. PATHOGENESIS OF ONYCHOMYCOSES: The mode of infection is still under debate. In many cases palmar and/or plantar tinea, exists but can often remain asymptomatic for years. After spreading to the nail, the fungus invades the hyponychium or lateral nail sulcus to finally reach the nail bed where it moves proximally to the matrix. Proximal subungual onychomycosis probably starts with a fungal skin infection, whereas white superficial onychomycosis seems to be a culture of T. mentagrophytes on a softened nail surface. Total dystrophic onychomycosis may result from both distal and proximal subungual onychomycosis or from C. albicans in chronic mucocutaneous candidiasis. Candida infections occur most often due to previous Candida paronychia, but it appears that a number of cases of so called idiopathic onycholysis are also caused by C. albicans with damage to the hyponychium being the portal-of entry. CLINICAL PICTURE OF ONYCHOMYCOSES: Onychomycoses can be divided into four different types. Distal subungual onychomycosis is the most common. The most frequent presenting clinical features are thickening and opaci-fication of the nail plate along the distal and lateral borders. The discoloration ranges from white to brown. The edge of the affected nail is usually uneven and often one or more streaks of dystrophic discoloured nail extend towards the distal border. Proximal subungual onychomycosis is uncommon. A white spot appears beneath the proximal nail fold and may extend distally to involve the deeper layers of the whole nail. Superficial white onychomycosis is also uncommon. The surface is the initial site of invasion. The causative organisms produce small superficial white and powdery patches over the nail. The surface becomes rough and the texture softer than normal. Total dystrophic onychomycosis represents the most advanced from all the previous three types, especially the distal subungual onychomycosis. The nail matrix has become permanently scarred by chronic infection. The nail is thick, elevated, denser and opaque. Candidomycotic onychomycosis shows erythematous and swollen proximal and lateral nail folds. Consequently, the nail plate becomes detached from the eponychium. Mycotic onycholysis is characterized by detachment of the nail plate from the bed, distal nail erosions, and grayish-yellow paste like material under the nail. DIAGNOSIS OF ONYCHOMYCOSES: The diagnosis of onychomycoses cannot be made on the basis of clinical observation alone. Direct microscopy plays an important role in diagnosing nail fungal infections. However, fungal cultures are the only definitive test that can be used to identify the genus and the species of the infectious organism. Histological examination is a routine technique useful for defining the nature and localization of fungi in the nail plate. Immunohistochemistry applied to onychomycosis is an experimental approach bringing prominent information about identification of fungi. In vivo confocal microscopy represents a technique of the future. PMID- 11432323 TI - [Osteoporosis]. AB - INTRODUCTION: Osteoporosis is a systemic skeletal disease characterized by low bone density and microarchitectural deterioration of bone tissue, leading to enhanced bone fracture risk. Due to its increasing prevalence (affects more than 10 percent of population) and consequences (bone fractures), osteoporosis is a growing medical, social and economic problem. PREVENTION AND STATISTICS: Of particular importance is hip fracture, with increasing incidence and lethal outcome in 20 percent, and less than 33 percent of patients with complete recovery. For better prevention of such undesired consequences, accurate diagnosis in the early phase of bone loss is necessary. Bone loss is a silent process, without signs and symptoms of the disease, and so active screening of persons at risk is needed, particulary among the postmenopausal women with some of the recognized risk factors for the development of osteoporotic bone fracture. In such persons, for early detection of bone mass loss, diagnostic measurement of bone density should be done, preferably dual-energy x-ray absorptiometry (DEXA). Prevention of osteoporosis is the best approach to the problem, and it should be started in early childhood with general measures (diet with sufficient calcium intake, life style with physical exercise) in a involved national program. CONCLUSION: Treatment of osteoporosis lasts for many years and includes general measures and medication with osteodensitometric assessment over a two-year period. PMID- 11432324 TI - [Children's knowledge of sex behavior and contraception]. AB - INTRODUCTION: Adolescent sexuality is not a new phenomenon, but it has been accepted differently at various times and societies. In modern times, adolescents start sexual life early, not well educated and not prepared for possible consequences. The period between the first sexual relationship, marriage and having children is prolonged with a greater possibility of having artificial abortions or sexually transmitted diseases (STD). It is of great importance to work on proper education of children and adolescents on sexual life and contraception. MATERIAL AND METHODS: An inquiry into the knowledge of sex and contraception was performed in an elementary school in Novi Sad, comprising 134 children from VII and VIII grade (aged 13-14, 77 girls and 57 boys) in order to prepare a lecture on these topics. RESULTS: Most of the knowledge on these topics children aged 13 and 14 got watching TV and reading magazines (44.15% girls and 70.17% boys) and from their friends (42.1%). Communication about sex and contraception exists mostly among friends (51.95% girls and 82.46% boys). One third of girls talk with parents and one quarter got knowledge from them. Only four boys (2.98%) had sexual intercourse without complications: artificial abortion or STD. Almost every child (96.95%) knows about AIDS and 89.25% children know about at least one method of contraception (mostly condom). Three quarters claim artificial abortion is harmful. 50% of children want more education about sex and contraception. DISCUSSION AND CONCLUSIONS: Most of the knowledge on sexual life and contraception children get from various forms of mass-media and from one another which is unreliable and incomplete. Unfortunately, parents and teachers play a minor role in sex education of children. In order to prevent spreading of sexually transmitted diseases and to decrease the percentage of adolescent pregnancy and artificial abortions, it is of great importance to investigate knowledge, attitudes and practice of children and adolescents in regard to sexual life and contraception and to prepare acceptable and efficient programs for education on these topics. However, not only education is important, but also acceptance and behavior of children and adolescents resulting from these programs. PMID- 11432325 TI - [The Stenger Test--its use and variations]. AB - INTRODUCTION: Simulation, dissimulation and aggravation of partial hearing loss can refer to bilateral or unilateral hearing loss. Tests for detection can be divided into unilateral and bilateral. CASE DESCRIPTION: A patient aged 29, came for otolaryngological examination 13 days after hearing impairment and blast injury of the ear following detonation of an airplane-launched missile about 500 meters away. AT the moment of detonation he was not wearing a helmet. Two hours later he noticed bloody discharge and strong tinnitus in the left ear. During examination and audiometric test, we suspected aggravation. The patient was sent for sub-specialist audiological examination. The audiogram made during the first testing showed a perceptive hearing impairment on the right side, while the left ear was without response (Audiogram 1). We asked the patient to concentrate, repeated the test, and got Audiogram 2 (Curve I). After this the patient was informed that we would carry out another testing, using Stenger test, and we got Curve II. The patient was informed that he had done very successfully in the test, and that we would repeat it, Curve III. We got Curve IV by repeating the test reducing the tone volume by 10 dB again. The aggravation was obvious, and the audiogram was shown to the patient, explaining that his intention was detected, and he was asked to cooperate correctly so that adequate therapy could be applied, which he accepted, so that we got Curve V, the real condition of his hearing. CONCLUSION: We have shown the case of a patient injured in a missile explosion which caused perforation of the tympanic membrane and a sensorineural hearing loss. Suspecting aggravation, Stenger test was performed. The testing showed unexpected results not described with Stenger test and we simultaneously detected the patient's intention to simulate aggravation. PMID- 11432326 TI - [Bacillus subtilis IP 5832 (Flonivin BS) shortens the duration of the carrier state in patients with acute non-typhoid salmonella enteritis]. AB - INTRODUCTION: The objective of this clinical trial was to assess the efficacy and safety of Flonivin BS (Bacillus Subtilis IP 5832) vs. placebo in treatment of patients with acute non-typhoid Salmonella gastroenteritis (ANSG), on adequate diet and parenteral rehydration therapy. MATERIAL AND METHODS: The clinical trial was conducted as a multicenter, controlled, randomized, double-blind, prospective clinical trial including 63 patients of both sexes, 20 to 52 years of age, with clinically, epidemiologically and microbiologically diagnosed ANSG, accompanied by diarrhea not longer than two days. RESULTS AND DISCUSSION: Flonivin BS capsules (each capsule containing at least 1 billion of vegetative spores of Bacillus subtilis, strain IP 5832) were administered to 31 patients, while 32 patients received placebo (3 x 2 capsules daily) over a seven day period. Stool cultures were collected on days 7, 14, 21 and 28 for checking the carrier state. The efficacy criteria were based on determining the differences in duration of diarrhea and of the carrier state in Flonivin BS-treated group of patients as compared to the placebo-group. Statistical analysis was performed using Student's test of proportions for small and large samples. The analysis of the number of patient-weeks by subgroups on days 7, 14, 21 or 28 of the trial in Flonivin BS- and placebo-groups has shown that the rate of patient-weeks of the carrier state was significantly lower in Flonivin BS-treated group as compared to the placebo group, which was manifested in week 4 of the trial (p < 0.01). The analysis of the rate of positive findings in stool cultures in the groups treated with Flonivin BS vs. placebo by weeks of the confirmed carrier state has shown that Flonivin BS significantly reduces the rate of positive findings in stool cultures in the fourth week from the initiation of treatment (p < 0.05). No differences were noted in duration of diarrhea between the study groups (p > 0.05). CONCLUSION: It has been concluded that a seven day treatment with Flonivin BS in patients with ANSG is of significance in prevention of a long-term carrier state, as it reduces the number of diseased that remain carriers in the third week after completion of treatment (p < 0.05). A prophylactic administration of Flonivin BS in patients with ANSG has a potential epidemiological significance due to a significant reduction in the number of patient-weeks of the carrier state in population of long-term carriers (p < 0.01). PMID- 11432327 TI - [Chronic venous insufficiency and compression therapy]. AB - INTRODUCTION: Disorders of venous circulation are among most frequent diseases in the human population. During recent years there has been increased interest in physiopathology of chronic venous insufficiency (CVI), due to development of more accurate diagnostic methods and new therapeutic techniques. Considering patophysiology of CVI and its consequences the crucial points are stasis and reflux of blood flow, with increase of intravascular pressure. CLASSIFICATION AND THERAPY OF CVI: Empirical experience and theoretical models suggest that progression of the disease can be stopped or reverted by correction of stasis. Today, this is the main direction in all therapeutic approaches. Compression therapy represents the most successful conservative technique. In this article we are presenting basic principles of the compression therapy of CVI. Evidence for the physiologic effect of compression therapy are decrease of edema, softening of lipodermatosclerosis, acceleration of venous flow, decrease in venous volume, blood shift into central compartments, reduction of venous refluxes, influence on arterial flow and improvement of microcirculation and lymph drainage. There are several types of compression devices: elastic stockings, elastic bandages, inelastic bandages, intermittent pneumatic compression devices. Other advantages of compression therapy are decrease of hospital treatment and better quality of life for patients with CVI. PMID- 11432328 TI - [The HLA antigen system in married couples with recurrent spontaneous abortions]. AB - INTRODUCTION: Investigations of factors responsible for the disturbance of fetomaternal tolerance during pathological pregnancies and causes of recurrent spontaneous abortions are pointing that HLA antigens play a key role in their development. Rejection of semiallogeneic allograft in this pregnancy disorder is a consequence of interaction between genetic and immunologic phenomena. MATERIAL AND METHODS: This article reports results of an investigation of human lymphocyte antigens in 130 couples with recurrent spontaneous abortions and in control group of 57 healthy couples. In both investigated groups the first class of human lymphocyte antigens was detected using microlymphocytotoxicity test by Terasaki. The phenotypic frequencies in the group with recurrent spontaneous abortions were compared with the corresponding results in the control group. We investigated the relative risk for development of this disorder and statistically significant difference of human lympocyte antigen frequencies between both investigated groups. RESULTS: The results of investigation showed that in the group of couples with recurrent spontaneous abortions the relative risk is hihger than 1 for the following human lymphocyte antigens: A2 = (1.53), A9 = (1.149), A10 = (1.07), A29 = (1.70), B18 = (2.24), B40 = (3.31). A significant difference in human lymphocyte antigen frequencies was established for HLA A2, HLA B18 and HLA B40, in comparison with the results of control group. Couples with recurrent spontaneous abortions had a statistically significantly higher rate of identical human lymphocyte antigens among spouses in comparison with the control group, at the level of significance p < 0.05 = 6.32. CONCLUSION: The results of this investigation showed that human lymphocyte antigens play an etiopathogenic role in development of recurrent spontaneous abortions and that some of these antigens are increasing susceptibility to pregnancy disorders. PMID- 11432329 TI - [Hereditary angioedema in organs of the head and neck as an indication for emergency tracheotomy]. AB - INTRODUCTION: Angioedema (angioneurotic edema) is often associated with urticaria, but edema is located deeper in the skin and mucous membranes. There are limited, painless, soft and medium hard swellings. Lack of general symptoms is evident, except if mucous membranes of the gastrointestinal system are affected and pain appears. It is particularly dangerous if located in the brain or larynx when there is a risk of suffocation. In 20% cases with laryngeal involvement intubation or tracheotomy is necessary. CASE REPORT: This is a case report of a patient hospitalized at the Clinic, having a swelling at the front side of neck, lower lip and difficulties with deglutition. Occasionally the patient had similar difficulties in the main joints whereas periodical swellings are characteristic for his father, sister, and sister's daughter. Clinical check up indicated a greyish swelling, of the oropharynx structure, with 1 cm respiratory space. Larynx was not visible due to swelling of epiglottis. The patient received intravenous steroid therapy, followed by infusion of physiologic solution with calcium. As his condition become very bad half an hour after admittance he was transported to the operation room. He received adrenaline but within the excepted time his respiration did not improve. Emergency tracheotomy was performed and afterwards his respiration and skin colour became normal. Regarding family anamnesis, clinical picture and laboratory results, hereditary angioedema was diagnosed. DISCUSSION: Hereditary angioedema is a rare form of angioedema which is an inherited autosomal dominant disorder. The disease is a result of deficit in C esterase inhibitor, which is a serum glycoprotein of SERPIN family (serum protease inhibitors), that is usually synthesized in hepatocytes. All the diseased are heterozygotes. There are two genetic variantions of the disease: I--patients with decreased quantity of inhibitor level in serum due to decreased synthesis and II--patients that have normal protein concentrations, but with abnormal protein, which is functionally inactive. Laryngeal edema can very soon cause narrowing of respiratory space, and if tracheotomy is not performed on time, suffocation occurs. Tracheotomy is one of the most urgent surgical interventions with the purpose to make patient's breathing easier to prevent suffocation and sometimes to save thr patient from certain death. CONCLUSION: In differential diagnosis of laryngeal edema, hereditary angioedema should be considered. Therapy of acute hereditary angioedema attacks should involve antihistamines, corticosteroids and adrenaline, as well as administration of fresh frozen plasma or infusion of C 1 inhibitor concentrate. Hereditary angioedema of the head and neck causing airway obstruction, is an indication for emergency tracheotomy. PMID- 11432330 TI - [Why aren't Leydig cell tumors of the ovary diagnosed in time?]. AB - INTRODUCTION: Leydig cell tumor is a rare sex-cord tumor with sex steroid secretion (mostly testosterone). All clinical symptoms and signs are the consequence of extremely high testosterone level. The diagnosis is confirmed using hormone analysis. CASE DESCRIPTION: A 46-year-old woman with previously normal reproductive function was admitted to the hospital due to hirsutism, temporal hair loss, hoarse voice, increased libido, amenorrhea and clitoromegaly. The examination revealed hirsutism score 36 and signs of defeminization and virilisation. Basal testosterone levels were extremely high before operation, while follicle stimulating hormone and luteinizing hormone were suppressed. Androstenedione was above normal level, while other hormones were normal. Testosterone increased twice in response to Pregnyl. In regard to dexamethasone test there was no response of testosterone, indicating a virilizing ovary tumor. The ultrasound of pelvis was normal. Intraoperatively, a solid 2.3 cm Leydig cell tumor was diagnosed. Testosterone decreased promptly. CONCLUSION: This study points to a long-term diagnostic procedure in Leydig cell tumor detection and effects of endogenous testosterone secretion on hypothalamic-pituitary-ovarian axis. PMID- 11432331 TI - [Awarding honorary membership in the German Society of Oromaxillofacial Surgery to Prof. Dr. Dr. h. c. mult. Hugo L. Obwegeser]. PMID- 11432332 TI - [Mechanical stimulation of osteoblasts in cell culture]. AB - BACKGROUND: Mechanical loading of bone is known to play a crucial role in bone remodeling and regeneration. Whereas the clinical effects of mechanically modulated bone healing have been extensively studied, less is known about the underlying mechanisms on a cellular level. This study was aimed at investigating the effects of uniaxial strains on osteoblast-like cells in culture. Mechanical loading was applied in physiological and hyperphysiological magnitudes. Nonstimulated cultures served as controls. RESULTS: Cultured primary bovine periosteal cells exhibited phenotypic features of osteoblast-like cells. Application of physiological strains (2,000 mu strain) led to a bone-specific expression of extracellular matrix proteins (osteonectin, osteocalcin, collagen type I). Hyperphysiological loads (10,000 mu strain) were associated with an increased synthesis of proteoglycans. Proliferation of cells was higher than the controls at 10,000 mu strain and showed no difference from physiologically loaded osteoblasts. DISCUSSION: Our study demonstrates that physiological loading of osteoblast-like cells enhances the regenerative capacity of bone, whereas hyperphysiological loads may impair bone regeneration. PMID- 11432333 TI - [Pneumatization of the zygomatic arch on pantomography]. AB - BACKGROUND: The pneumatized spaces of the zygomatic arch (zygomatic air cell defect) are a clinically asymptomatic variation of the temporal bone. The empty spaces of the articular eminence and temporal bone of the zygomatic arch are depicted on radiographs as a sharply demarcated osteolytic lesion of round or oval shape. The aim of this study was to determine the prevalence and characteristics of the ZACD in our outpatients. MATERIALS AND METHODS: We investigated the panoramic radiographs of 1,084 patients who were treated during 1 February and 31 July 2000. The mean age of the 1,084 patients was 41.9 years (SD: 17.2 years; range: 2-96): 628 patients were male (58%) and 456 female (42%). A ZACD was found in 20 of 1,084 patients (prevalence: 1.85%). The mean age of the patients with a ZACD was 43.2 years (SD: 18.4 years; range: 7-87): 11 were female (55%) and 9 male (45%). RESULTS: A bilateral ZACD was found in 20% (four patients). Interestingly, one of these ZACD patients was a 7-year-old boy. Up to now, it had generally been accepted that pneumatization of the zygomatic arch takes place after puberty with the youngest persons with ZACD being 15 years of age. Our findings should prompt consecutive studies on the prevalence of ZACD in children. This finding can be relevant in patients treated for spreading otitis media or mastoiditis. PMID- 11432334 TI - [Cortical thickness of the mandible with special reference to miniplate osteosynthesis. Morphometric analysis of autopsy material]. AB - Thirty-two human mandibles were marked with three typical fracture lines: a low condylar fracture, a fracture of the mandibular angle, and one through the mental foramen on each side. The mandibles were sectioned at the fixation sites of the miniplate screws. The thickness of the cortical layer was measured with a scaled magnifying glass at the points of anthropological interest and at the marked screw holes. The inferior cortical layer turned out to be thickest in the anterior area. In contrast, it was very thin at the mandibular angle, which might explain the higher complication rate in treating fractures in this region. The thickness of the lingual cortex increased up to the symphysis, whereas the buccal cortical layer showed a decline in size from the mandibular angle up to the chin. The cortical bone at the alveolar ridge was porous. The cortical supply for miniplate osteosynthesis at the condylus ranged from 1.1 mm up to 1.74 mm, which seems to be limited, but due to the small diameter of the condylus most common screws obtain additional anchorage in the inner cortex. The thickness of the cortical layer at the mandibular angle increased from 1.47 mm at the ramus up to 1.97 mm at the beginning of the corpus, reaching 2.14-2.38 mm for the lower plate at the mental foramen. The results for the upper plate were slightly lower. PMID- 11432335 TI - [Magnetic resonance tomography for planning dental implantation]. AB - PROBLEM: Three-dimensional imaging diagnostics are increasingly recommended before inserting dental implants in high-risk areas and in cases of severe alveolar atrophy. Since patients are exposed to considerable radiation with computed tomography (CT), the possibilities of employing magnetic resonance imaging (MRI) of the jaw as a diagnostic imaging method before inserting dental implants were examined. MATERIAL AND METHOD: Twelve patients and three volunteers were examined by MRI with T1-weighted, fat-suppressed sequences and conventional T1-weighted sequences. The patients wore a diagnostic splint including markers- in the form of capillaries filled with 0.025 x 10(-2) M gadolinium solution (1.5 mm in diameter)--in the planned implant's position and axis. RESULTS: The presentation of relevant anatomic structures and the three-dimensional accuracy of the markers were judged. Metal artefacts were evaluated in vitro. The MRI of the jaw and midface represents the mandibular canal, the maxillary sinus, and other decisive anatomic structures by detailed representation of the connective tissue surrounding the bone. Artefacts of metallic fillings reduce the image quality. CONCLUSION: Obtaining clinical findings and planning before inserting dental implants with the help of MRI can certainly be applied with toothless patients and facilitates three-dimensional planning by representing the exact location and angle of the drill tubes. Local restrictions result from metal extinction artefacts in jaws with teeth and in controls after having inserted titanium implants. PMID- 11432336 TI - [Life threatening and fatal complications of radical neck dissection]. AB - BACKGROUND: Between 1990 and 1999, 395 neck dissections were performed in 357 patients: 195 left-sided (105 of these radical) and 200 right-sided (107 of these radical). Life-threatening complications occurred in four cases and two patients died. CASE REPORTS: After left-sided radical neck dissection with chylous fistula, a chylothorax resulted, which could not be controlled in spite of chest tube drainage and thoracotomy so that the 75-year-old female patient died 30 days postoperatively. A 66-year-old man died 35 days after a functional neck dissection of the left side because of extreme chylous flow of up to 7 l/day in spite of parenteral nutrition, local surgical revision, and intrathoracic ligation of the thoracic duct. Undetected cirrhosis of the liver was regarded to be the reason for this extremely increased lymph flow. In a 63-year-old man, a jugular foramen hemorrhage during radical neck dissection could only be managed after 3 h and approximately 6 l of blood loss. In a 66-year-old man, a discrete injury of the pleura led to cardiovascular failure due to a tension pneumothorax with mediastinal shift about 45 min later, which required immediate chest tube placement. DISCUSSION: In none of these unusual cases, which accounted for 1% of all evaluated interventions, had the patients been informed about the observed complication. However, no legal consequences resulted. Nevertheless, dramatic courses of chylous fistulas and rare complications should be considered both forensically when seeking informed consent and clinically. PMID- 11432337 TI - [Resorbable osteosynthesis material in craniosynostosis]. AB - BACKGROUND: The results of using resorbable plates and screws (82% polylactic acid and 18% polyglycolic acid) in craniofacial surgery for the correction of craniosynostosis after more than 4 years of experience are presented. Special attention is focussed on the degree of stability and the clinical tissue response to the material employed to answer the question of whether the material is an adequate alternative to titanium. METHODS: Thirty patients who had been treated with this method for craniosynostoses were examined at regular intervals regarding the shape and stability of the forehead region, visibility and palpability of the plates, and tissue reactions. RESULTS: The technical handling of the osteosynthesis material proved to be simple and reliable. In one case the bone was not strong enough for the screw pitch. After an observation period of a maximum of 4 years and 1 month, the fixations were stable with no signs of adverse reactions. DISCUSSION: If the long-term results remain favorable, we consider the use of resorbable material a promising method for the stabilization of segments in craniofacial surgery in children. PMID- 11432338 TI - [Focal palmoplantar and oral mucosa hyperkeratosis syndrome]. AB - CASE REPORT: Four members of a family in three generations are presented who were affected by a rare syndrome (mucosa hyperkeratosis syndrome). This syndrome is characterized by autosomal-dominant inheritance, white lesions of the gingiva, and palmoplantar hyperkeratosis. The four affected members of the family revealed an abnormal keratinization of the gingiva and palmoplantar epidermis. Biopsies of plantar and gingival lesions histologically showed acanthosis and hyperkeratotic cornification of the epithelium. Electron microscopy demonstrated the features of epidermolytic hyperkeratosis. DISCUSSION: From the differential diagnostic point of view, the mucosa hyperkeratosis syndrome has to be distinguished from the Jadassohn-Lewandowsky syndrome and the Howel-Evans' syndrome, which is associated with esophageal carcinoma. PMID- 11432339 TI - [Psycho-oncology and its scientific background]. AB - Psycho-oncology has two purposes corresponding to the psychosocial aspects of cancer: 1. to clarify the psychosocial impacts of cancer on the quality of life of the patient, family and staff; 2. to clarify the role that psychosocial and behavioral variables may have in cancer risk and survival. For these purposes, patient education, counseling, behavioral and psychopharmacological techniques have been applied to clinical oncology. In this paper, we review psycho-oncology and its scientific background, with respect to neuroradiology and molecular genetics, as well as psychiatry, psychology, epidemiology, pharmacology and immunology. PMID- 11432340 TI - [Circulating cancer cells in the peripheral blood]. AB - Occasional observations on this subject were seen until 1995 when Engell reported the large study on circulating cancer cells in the peripheral blood. Engell's report generated great enthusiasm in our country also. But, these cytologic method have a low sensitivity. Consequently, studies on the circulating tumor cells have been abandoned. However, since 1987, RT-PCR and other molecular biologic method were introduced for the identification of circulating cancer cells and micrometastases in bone marrow as well as in lymphnode. Serial analysis of a large number of patients by well-qualified laboratories have been performed. Several authors have shown a significant correlation between detection of cancer cells and relapse in some forms of malignancies. These research may be beneficial in the staging of cancer, in monitoring the effectiveness of cancer treatment, etc. Detection of cancer cells in the peripheral blood likely to have a profound impact on the practice of clinical oncology. PMID- 11432341 TI - [The clinical significance of detecting small numbers of carcinoma]. AB - The clinical significance of detecting small numbers of carcinoma cells has been emphasized. The techniques for detecting micrometastasis in the bone marrow, lymph node, and peripheral blood have progressed remarkably in recent years with the advance of molecular biology. We use molecular biological techniques, such as reverse transcribed-polymerase chain reaction (RT-PCR) targeting the carcinoembryonic antigen (CEA) gene, and magnetic-activated cell separation system (Miltenyi Biotec Inc., CA) plus modified MASA targeting for K-ras mutation in practical clinical diagnosis and treatment. We herein, we describe how to obtain samples, such as bone marrow, lymph node, and peripheral blood, and how to prepare these samples. We then discuss the pros and cons of tumor specificity and detectability in several methodologies. Thereafter, we describe our strategies with regard to RT-PCR for CEA mRNA and MACS plus MASA method for K-ras mutations. At present, however, we have no consensus for anyone method, and therefore, stress the need to establish an appropriate methodology through a cooperative study. PMID- 11432342 TI - [The minimal residual disease (MRD) in hematological malignancies]. AB - Molecular genetic and cytoimmunological markers have been applied for the detection of minimal residual disease (MRD) in hematological malignancies. These markers include surface markers or rearranged T-cell receptor and immunoglobulin genes in the lymphoid malignancies and fused genes associated with chromosomal translocations such as BCR-ABL in t(9;22) or PML-RAR alpha in t(15;17) in myeloid malignancies. The expression of the WT1 gene is recognized as the universal tumor marker for a wide variety of hematological malignancies. Using these sensitive markers, a tumor cell in 10,000 to 1,000,000 normal cells can be detected. By examining a large number of patients, it has been shown that the MRD in the early phase of chemotherapy has a correlation with the prognosis of childhood ALL. Based on these observations, a new strategy of chemotherapy in which the post remission therapy is modified based on the MRD results has begun. The amount of tumor cells contaminated in the autologous stem cell grafts in ALL patients might be related to the prognosis. The diagnosis of MRD will be used as an important routine examination in chemotherapy for leukemia/lymphoma patients. PMID- 11432343 TI - [Clinical significance of detecting circulating cancer cells in patients with solid malignancies]. AB - There have been several studies attempting to detect the existence of cancer cells in the peripheral blood of patients with solid malignancies. The use of RT PCR assays for cytokeratin (CK), carcinoembryonic antigen (CEA), alpha fetoproteins (AFP), prostate specific antigen (PSA) and prostate specific membrane antigen (PMSA) is likely to be practicable in the detection of circulating tumor cells from epithelial-derived malignancies. The positive rates varied widely among the studies attempting to detect circulating cancer cells in peripheral blood, even when the same targets were used, which may be explained by the sensitivity of the RT-PCR assays and the target genes used. Both false negative results and false positive results can be obtained with the RT-PCR assay system. Furthermore, cancer cells may be released from the primary cancer into the circulation intermittently rather than constantly, and the results may vary among the samples obtained at different time points. In this report, we review our recent studies and related studies by other researchers to show the advances and the problems in detecting circulating cancer cells in patients with solid malignancies. The clinical significance of detecting circulating cancer cells in peripheral blood remains to be determined. PMID- 11432344 TI - [Clinical significance of micrometastasis to lymph nodes in gastrointestinal tract cancers]. AB - Surgery is the main therapy for malignancies of the gastrointestinal tract. Lymph node metastasis is one of the major factors in predicting patients' clinical course and choosing appropriate adjuvant therapy after surgery. The concept of micrometastasis to regional lymph nodes emerged over 10 years ago, but its significance has been controversial. To clarify the relevance of micrometastasis of gastrointestinal tract cancers, we have established RT-PCT based-diagnostic methods using multi-markers such as CEA, CK20, and Mage 3. Prospective studies have shown that not a few micrometastasis-positive patients with carcinoma of the colon, stomach, and esophagus suffered disease recurrence, even though they did not show histologically positive lymph node metastasis. They were initially diagnosed as node-negative, and thus predicted to be disease free. A retrospective study of 62 patients with stage II node-negative colorectal cancer showed that 5-year overall survival was 78.2% among micrometastasis-positive patients, against 95.3% micrometastasis-negative patients. Moreover, there was a marked difference in 5-year disease-free survival, with 61.4% versus 88.4%, respectively. These data warrant further prospective study with a large population since RT-PCR based detection systems for micrometastasis appear to have the potential to improve conventional diagnosis and therapy for colorectal cancer. PMID- 11432345 TI - [Rapid quantitative detection of free cancer cells in the peritoneal cavity of gastric cancer patients with real-time RT-PCR, and its prognostic significance]. AB - We have established a rapid and quantitative detection method using real-time RT PCR on the LightCycler instrument. This method can reproducibly quantitate 10 10(5) CEA expressing colon carcinoma cells per 10(7) peripheral blood leukocytes, a sensitivity comparable with conventional RT-PCR with a wide linear measuring range. Analysis of peritoneal washes from 241 gastric cancer patients with this assay revealed relative values of CEA transcripts that correlate well with the depth of tumor invasion. The sensitivity and specificity of real-time RT-PCR with a cut-off value of 0.25 were 88% and 81%, respectively. At this cut-off value, patients in the real-time RT-PCR (+) group were found to suffer from peritoneal metastasis with a high frequency, while peritoneal recurrence was very rare among patients with real-time RT-PCR (-) results. These results indicate a positive correlation between CEA mRNA levels in peritoneal washes and patient prognosis. We conclude that real-time RT-PCR with hybridization probes is a sensitive, quantitative, specific and rapid method to detect free cancer cells in peritoneal washes. PMID- 11432346 TI - [Multi-institutional cooperative study on combination chemotherapy with THP, CDDP and 5-FU for squamous cell carcinoma of the head and neck]. AB - Combination chemotherapy with THP, CDDP and 5-FU for squamous cell carcinoma of the head and neck was conducted in 13 institutions in Hyogo Prefecture as a multi institutional cooperative study. In the initial study (Nov. 1990-Nov. 1993), THP was administered intravenously at 20 mg/m2 on day 1, CDDP at 80 mg/m2 on day 2, and 5-FU at 1,000 mg/body/day in a continuous drip infusion for 120 hours from day 2 to day 6. In the second study (May, 1996-Mar. 1998), THP was administered at 20 mg/m2 on day 1, 5-FU at 10 mg/kg/day from day 1 to day 5, and CDDP at 70 mg/m2 on day 6 in the same way as the initial study. Forty-nine patients (Stage I in 3, Stage II in 12 including 2 recurrent cases, Stage III in 6, Stage IV in 28 including 3 recurrent cases; 1 course chemotherapy in 13 and 2 or more courses in 36) were subjected as complete cases in the initial study, and 36 patients (Stage I in 5 including one recurrent case, Stage II in 11 including 1 recurrent case, Stage III in 9 including 2 recurrent cases, Stage IV in 11 including one recurrent case; 1 course in 18 and 2 or more courses in 18) in the second. The overall response rate was 65.3% (CR in 3 cases) in the initial study and 63.9% (CR in 5 cases) in the second. Primary cases showed a response rate of 65.9% (29/44) in the initial study and 71.0% (22/31) in the second, whereas recurrent cases showed a 60.0% (3/5) response rate in the initial study and a 20.0% (1/5) rate in the second. Treatment-naive patients showed a response rate of 72.7% (24/33) in the initial study and 71.0% (22/31) in the second, whereas previously treated patients showed a 50.0% (8/16) response rate in the initial study and a 20.0% (1/5) rate in the second. Adverse reactions of more than Grade 3 in the initial study were leukopenia in 18.4%, thrombocytopenia in 8.2%, decrease of hemoglobin in 6.1%, loss of hair in 6.1%, anorexia in 36.7%, nausea and vomiting in 26.5%, and diarrhea in 4.1%, whereas those of Grade 3 in the second study were decrease of hemoglobin in 2.8%, anorexia in 22.2% and nausea and vomiting in 8.3%. From these results, it is suggested that the regimen in the second study was more useful than that in the initial study. PMID- 11432347 TI - [Efficacy of weekly docetaxel therapy for advanced or recurrent breast cancer]. AB - To evaluate the safety and efficacy of weekly docetaxel (weekly TXT) in cases of advanced or recurrent breast cancer, 31 patients were enrolled in this pilot study of weekly TXT given at 25 mg/m2/w. Each cycle consisted of 3 weeks of therapy followed by a 1-week treatment break in an outpatient setting. Patients received a median of 15 infusions with a median cumulative dose of 680 mg. The median time to treatment failure was 8 months. The overall response rate was 32.3%, and 22.6% of patients had stable disease for at least 6 months. The response rate was consistent regardless of prior chemotherapy with anthracycline. There was no grade 3 or 4 toxicity, and the regimen was generally well tolerated. Although 37.5% of patients had grade 1 or 2 nail change, myelosuppression, fatigue, nausea, vomiting and fluid retention were mild. Weekly TXT seems to be an effective and feasible treatment for advanced or recurrent breast cancer patients. PMID- 11432348 TI - [Inhibition of HCFU absorption after resection for gastric cancer--application of hydroxyaluminium gel]. AB - HCFU (carmofur; Mifurol) is an 5-FU analog. The maximum blood concentration of HCFU in HCFU fraction (Cmax) after gastric resection was higher than before resection. Hot sensation and pollakiuria, characteristic side effects of HCFU, are dependent on concentration of HCFU fraction in blood. Therefore, it is considered that the frequency of occurrence of side effects after gastric resection is high. For that reason, we thought that if absorption of HCFU could be reduced, fewer side effects would result. We focused on the fact that drugs which include aluminium gel may decrease absorption in combined drugs, and thought it would be possible to delay absorption in HCFU by using them. We studied the HCFU concentration in the HCFU fraction and 5-FU concentration in blood, respectively, in two cases: 1) single oral administration of HCFU 100 mg and 2) coadministration with hydroxyaluminium gel (ALG) 10 ml in the whole or partial resection of gastric cancers for 8 patients. We found that the concentration of HCFU in the HCFU fraction 2 hours after its administration decreased significantly: 3.24 +/- 1.78 (single administration), 1.37 +/- 0.91 (coadministration with ALG) (p = 0.023). HCFU concentration in the HCFU fraction seemed to decrease for coadministration with ALG in the area under the time-blood concentration curve (AUG) (p = 0.071). The 5-FU concentration did not seem to decrease in either case. From these results, the coadministration of HCFU with ALG seems to be effective for the inhibition of adverse drug reaction after the resection of gastric cancers. PMID- 11432349 TI - [Tumoral levels of thymidine phosphorylase and dihydropyrimidine dehydrogenase in elderly colorectal cancer patients]. AB - Fluoropyrimidine therapy for elderly colorectal cancer patients remains controversial. Tumoral levels of thymidine phosphorylase (TP), dihydropyrimidine dehydrogenase (DPD), and the ratio of TP to DPD determined by an enzyme-linked immunosorbent assay were compared between colorectal cancer patients aged 75 or over (elderly group, n = 25) and those 74 years or less (control group, n = 87), in order to examine the characteristics of colorectal cancers in the elderly from the viewpoint of metabolic and anabolic pathways of fluoropyrimidines. The level of TP was 78.4 +/- 47.0 unit/mg protein in the elderly group and 82.4 +/- 70.9 unit/mg protein in the control group (p = 0.86). The level of DPD was 53.7 +/- 43.1 unit/mg protein in the elderly group and 52.6 +/- 37.7 unit/mg protein in the control group (p = 0.73). The ratio of TP to DPD was 2.0 +/- 1.2 in the elderly group and 1.8 +/- 0.9 in the control group (p = 0.44). These three parameters did not differ between the groups when divided according to Dukes' stage (Dukes' A.B versus Dukes' C.D). These results suggest that there are no age specific characteristics in relation to conversion of fluoropyrimidines such as capecitabine and doxifluridine to 5-fluorouracil (FU) and degradation of 5-FU in colorectal cancers. PMID- 11432350 TI - [Effect of G-CSF (nartograstim) on neutropenia (leukopenia) induced by taxane in metastatic breast cancer--time-course changes in neutrophil and leukocyte counts]. AB - Since 1997, we have used docetaxel and paclitaxel as the second-line and third line chemotherapies against anthracycline-resistant metastatic breast cancer. However, these taxane compounds induced neutropenia and leukopenia, which may be reversed by G-CSF (Nartograstim). We thus examined the therapeutic efficacy of nartograstim for time-course changes in neutrophil and leukocyte counts in these patients. No difference was observed in neutrophil or leukocyte count whether the patient was treated with docetaxel or paclitaxel. Neutrophil and leukocyte counts reached a nadir on days 7 to 8 after administration. With a 5-6 day administration of nartograstim, neutrophil or leukocyte counts recovered by the second or third day after the nadir, indicating that the chemotherapy was given safely with nartograstim. In these same patients receiving a given treatment cycle, the number of days until reaching the nadir were almost identical for neutrophils and leukocytes; however, the duration of the nadir and the time to count recovery was significantly longer for neutrophils than for leukocytes. In the clinical setting, the parameter "leukocyte count" has been occasionally used for evaluation of the severity of myelosuppression, because the data is more readily available. However, at least during the nadir, the "neutrophil count" should be used as the parameter of choice. PMID- 11432351 TI - [A case of advanced gastric cancer treated with neoadjuvant chemotherapy of low dose CPT-11 + CDDP]. AB - Side effects due to administration of anti-cancer drugs often cause the treatment to be abandoned or a decrease in the amount of anti-cancer drugs. Recently, the anti-tumor effects of "low-dose CPT-11", which can be administered at the outpatient clinic, are reported. We performed "low-dose CPT-11 + CDDP" as a neoadjuvant chemotherapy to a patient with advanced gastric cancer. CPT-11 and CDDP combination chemotherapy caused very few side effects, so we could continue the treatment and achieve anti-tumor effects. Consequently, surgery could be performed, but disseminated metastasis was found so that the surgery ended as a non-curative operation. However, it was considered that this method of "low-dose CPT-11 + CDDP" was very effective as the neoadjuvant chemotherapy in a patient with advanced gastric cancer. PMID- 11432352 TI - [Clinical experience of intermittent administration of 5-FU and CDDP to patients with advanced and recurrent gastric cancer]. AB - We administered 5-FU and CDDP (FP) intermittently to four patients with advanced and recurrent gastric cancer. A minor response (MR) and partial response (PR) were achieved in two of four patients who had evaluable lesions for this treatment, and few side effects were observed. Only one patient among six showed grade 2 leucocytopenia after 15 administrations of this chemotherapy, when she had attained a partial response in a lung metastasis. The other patients had no side effects such as bone marrow suppression or digestive symptoms. This intermittent FP treatment may be an effective and promising therapy with few side effects even for the patients with serious conditions. PMID- 11432353 TI - [A case of esophageal carcinoma with multiple liver metastases successfully treated with nedaplatin (NDP) and 5-fluorouracil (5-FU) combined with radiotherapy]. AB - A 78-year-old male patient had esophageal carcinoma with multiple liver metastases. Chemoradiotherapy was performed. The chemotherapy consisted of protracted infusion of 5-fluorouracil (5-FU), combined with infusion of nedaplatin (NDP). Radiation of the mediastinum was administered concomitantly with chemotherapy. The patient showed a complete response (CR) of the primary lesion and a partial response (PR) of the liver metastasis for 11 months. Since liver metastasis recurred after initial treatment, chemotherapy consisting of NDP infusion combined with vindesine sulfate (VDS) infusion was performed. The patient again showed PR. Grade 3 leukocytopenia occurred during treatment, but there were no major toxicities such as thrombocytopenia, nausea, renal dysfunction or esophagitis. Survival time was one year and 7 months. In conclusion, concurrent chemoradiotherapy including NDP is effective and safe for patients with esophageal carcinoma accompanied by multiple liver metastasis. This nonsurgical approach may be an option for standard care in such cases. PMID- 11432354 TI - [A case of adrenal metastasis of lung cancer treated by carboplatin and docetaxel]. AB - A 42-year-old male was referred to our hospital in October 1998, suffering from severe cough accompanied by repeated hemosputa. Serum NSE slightly increased and cytology of sputum indicated class V. Chest X-P and CT revealed a large tumor in the right upper lobe and hilar lymphadenopathy. Abdominal CT revealed bilateral adrenal tumors. For continuous bloody sputum, a right upper lobectomy and lymphadenectomy were performed and the pathologic diagnosis was large cell carcinoma. After surgery, we chose radiation and chemotherapy. The new chemotherapeutic agent docetaxel (60 mg/m2 in combination with carboplatin (CBDCA: AUC 6,800 mg/m2) was administered, resulting in the remarkable reduction in the size of adrenal metastasis by 50% after 3 courses of chemotherapy. Furthermore, 12 months later, the right adrenal metastasis was remarkably reduced (5 x 3 cm-> 0.5 x 1.0 cm), and the left adrenal metastasis had disappeared on abdominal CT. These findings may suggest the efficacy of newly developed docetaxel in cases of non-small cell lung cancer. PMID- 11432355 TI - [A case of breast cancer with multiple bone metastases improved by high-dose toremifene]. AB - A 63-year-old woman underwent modified radical mastectomy with 3 cycles of adjuvant chemotherapy (cyclophosphamide, epirubicin, 5-fluorouracil) and MPA endocrine therapy for breast cancer. Because of nausea and general fatigue, she refused to continue this therapy and did not visit the hospital. When she came our hospital and 16 months later, she had developed multiple bone metastases. At the same time, she was suffering from lung tuberculosis. She was treated with toremifene at a dose of 120 mg/day without any side effects. After 3 months administration of toremifene, pain disappeared and her high serum CA15-3 and BCA225 dropped to within the normal range. On bone scintigrams, abnormal accumulation almost disappeared after 9 months of administration of toremifene. In this case, the patient was suffering from lung tuberculosis and did not desire intensive chemotherapy. Administration of high-dose toremifene was effective for multiple bone metastases without any side effects. PMID- 11432356 TI - [A case of papillary adenocarcinoma of the ovary that responded favorably to irinotecan hydrochloride and cisplatin after the administration of paclitaxel and carboplatin]. AB - Recently, the standard treatment for advanced ovarian cancer has changed from CP therapy (cyclophosphamide, cisplatin (CDDP)) to TJ therapy (paclitaxel (TXL), carboplatin (CBDCA)). Irinotecan (CPT-11) is one of the derivatives of camptotecin and has been reported to have a high efficacy for ovarian cancer. In one case of ovarian cancer, chemotherapy was applied with CBDCA and TXL. However, after 2 months of six courses of the chemotherapy, CA-125 was elevated. The elevation of tumor marker levels in serum without the recurrent focus forced us to treat the patient with CPT-11 and CDDP for the second line chemotherapy. Tumor marker levels improved at the beginning of the therapy. In conclusion, CPT-11 and CDDP was effective against the recurrence of ovarian cancer treated with TJ therapy. PMID- 11432357 TI - [A case of retroperitoneal malignant fibrous histiocytoma with marked response to cisplatin, ifosfamide and doxorubicin]. AB - A 61-year-old man was admitted to our hospital in December 1994 for a suspected retroperitoneal tumor. Systemic imaging investigations demonstrated retroperitoneal solid tumor, which was diagnosed as malignant fibrous histiocytoma (MFH) by immunohistochemistry for alpha 1-antitrypsin. In March 1995, he was treated with 3 courses of systemic chemotherapy with cisplatin, ifosfamide and doxorubicin followed by the same therapy in March 1996, without serious side effects. MFH is known to be resistant to ordinary chemotherapy. However, the CT showed a marked decrease in the size of the tumor, and the tumor disappeared within 2 months after the first treatment. The patient also recovered rapidly from abdominal pain, for which complete remission has been achieved for more than 5 years. The present chemotherapy may be an effective treatment for retroperitoneal MFH. PMID- 11432358 TI - [New oral anticancer drug, TS-1 (S-1)--from bench to clinic]. AB - We describe in this paper a therapeutic modality which is based on a self rescuing concept (SRC) featuring dual activity, i.e., effect-enhancing activity and adverse reaction-reducing activity. We present the theory and practice of S 1, a novel oral fluoropyrimidine anticancer agent designed to enhance anticancer activity and reduce gastrointestinal toxicity through the deliberate combination of the following components: an oral fluoropyrimidine agent, tegafur (FT); a DPD inhibitor (CDHP: 5-chloro-2, 4-dihydroxypyridine) which is about 200-fold more potent than uracil used in UFT; and an ORTC inhibitor (Oxo: potassium oxonate) which is localized in the gastrointestinal tract. We devised a novel oral anticancer agent, S-1, as a combination drug with a molar ratio of 1:0.4:1 for FT, CDHP, and Oxo, respectively. To compare S-1, FT, and UFT in terms of their anticancer activity and adverse reactions, a colon cancer implantation model in rats was used for 4-week consecutive oral administration from the time when the postimplantation tumor weight become about 2 g. The tumor disappeared on day 16 at a given dose of S-1 (as 22.5 mg/kg FT), and the tumor did not reappear for at least three months. Antitumor activity was more marked with S-1 than FT and UFT. Adverse reaction, i.e., stomatitis, depilation, and weight loss, were less frequent in the S-1 group than in the other groups. A clinical pharmacology study examined blood concentrations of 5-FU after twice-a-day administration after meals of S-1 at a dose of 40 mg/m2. Blood concentrations of 5-FU were 60 to 200 ng/ml in all twelve patients examined. Late phase II clinical trials of S-1 were conducted in patients with advanced and recurrent stomach cancers, in the same regimen as for the clinical pharmacology study. It basically consisted in four cycles, each of which comprised 4-week, twice-a-day, consecutive oral administration with a 2-week withdrawal. The overall response rate was 44.6% (45/101). Median survival time (MST) was 224 days. S-1 was given manufacturing approval by the Ministry of Health and Welfare of Japan after a priority review, with indications for advanced and recurrent stomach cancers. A late phase II clinical study of S-1 in patients with advanced/recurrent head and neck cancer was conducted in 59 eligible patients. Objective responses were 4 complete response (CR) and 13 partial response (PR), for a response rate of 28.8% (17/59). MST was 344 days. Grade 4 hemoglobin decrease was observed in one case; however, this returned to normal after the termination of drug administration and blood transfusion. Therefore, this event was confirmed to be reversible. A late phase II clinical trial of S-1 was conducted to evaluate the efficacy and toxicities in patients with metastatic colorectal carcinoma. Sixty-three patients with measurable metastatic colorectal carcinoma were enrolled in this clinical trial. The overall response rate was 35.5% (22/62), and the MST was 378 days. The main adverse reactions were myelosuppression and GI toxicities. The incidence of neutropenia (Grade 3 or 4) was 13%, while the incidence of other adverse reactions was 10% or below. None of 53 outpatients required to be hospitalization due to adverse reactions. Late phase II clinical trials of S-1 are in progress for colorectal cancer, breast cancer and non-small cell lung cancer. To establish the standard therapeutic modality for cancers, including gastrointestinal cancers, in Japan, the conduction of clinical trials combining S-1 and other anticancer drugs holds promise for the future. PMID- 11432359 TI - [Pancreatic and biliary tract cancers]. AB - There are several tumor markers for pancreatic and biliary tract cancers, such as carcinoembryonal antigen, pancreatic enzyme, carbohydrate antigen and tumor associated gene. CA19-9 and SPan-1, which are type I carbohydrate antigens, are especially useful among these tumor markers. Combination assays of these tumor markers improve the positive rate for these cancers. These tumor markers are useful not only in diagnosis but also in assessing the therapeutic efficacy and early detection of recurrence after operation. PMID- 11432360 TI - Cancer services. Left to chance. AB - Most primary care groups/trusts have cancer lead posts and have some involvement in planning and commissioning cancer services. Cancer is not a high priority in comparison to other national service frameworks and the transition to PCT status. PCG/Ts want help and information about developing cancer services but not all want this now. Most PCG/Ts have some involvement in cancer networks but information needs exist about their role and potential. PMID- 11432361 TI - First person. My mum, my dad, my daughter and the NHS. PMID- 11432362 TI - Mental health. Reach out and join up. AB - The NHS plan proposed the establishment of 220 specialist assertive outreach teams for people with serious mental illness by 2003. Research on the development of these teams in London shows that diverse models are in operation. Some focus on medication administration, others on psychosocial needs. It is important that these teams are incorporated into the spectrum of care services. PMID- 11432363 TI - International recruitment. Learning the hard way. PMID- 11432365 TI - [Self-renewal and differentiation of ES cells]. PMID- 11432364 TI - Visual responses of ganglion cells of a New-World primate, the capuchin monkey, Cebus apella. AB - 1. The genetic basis of colour vision in New-World primates differs from that in humans and other Old-World primates. Most New-World primate species show a polymorphism; all males are dichromats and most females trichromats. 2. In the retina of Old-World primates such as the macaque, the physiological correlates of trichromacy are well established. Comparison of the retinae in New- and Old-World species may help constrain hypotheses as to the evolution of colour vision and the pathways associated with it. 3. Ganglion cell behaviour was recorded from trichromatic and dichromatic members of a New-World species (the capuchin monkey, Cebus apella) and compared with macaque data. Despite some differences in quantitative detail (such as a temporal response extended to higher frequencies), results from trichromatic animals strongly resembled those from the macaque. 4. In particular, cells of the parvocellular (PC) pathway showed characteristic frequency-dependent changes in responsivity to luminance and chromatic modulation, cells of the magnocellular (MC) pathway showed frequency-doubled responses to chromatic modulation, and the surround of MC cells received a chromatic input revealed on changing the phase of heterochromatically modulated lights. 5. Ganglion cells of dichromats were colour-blind versions of those of trichromats. 6. This strong physiological homology is consistent with a common origin of trichromacy in New- and Old-World monkeys; in the New-World primate the presence of two pigments in the middle-to-long wavelength range permits full expression of the retinal mechanisms of trichromatic vision. PMID- 11432366 TI - [Roles for FGF-FGFR signaling during vertebrate development]. PMID- 11432367 TI - Normothermic cardiopulmonary bypass is beneficial for cognitive brain function after coronary artery bypass grafting--a prospective randomized trial. PMID- 11432368 TI - Mechanical valves in tricuspid position: cause of thrombosis and prevention. PMID- 11432369 TI - SNFs--still a safe bet? Interview by Mark Zacharia. PMID- 11432370 TI - Tissue renin-angiotensin system: its expression, localization, regulation and potential role in the pancreas. AB - The classical concept of the renin-angiotensin system (RAS) is that of a blood borne cascade, whose final and bioactive product, angiotensin II, plays an important endocrine role in the maintenance of blood pressure and electrolyte as well as fluid balance. In addition to this circulating RAS, there are an increasing number of studies to suggest the existence of a local angiotensin generating system in several tissues. The so-called tissue RAS can act locally as a paracrine and/or autocrine factor in meeting specific needs for individual tissues and it can operate, in whole or in part, independently of the circulating counterpart. Recent studies on the expression and localization of key RAS components, particularly angiotensinogen and renin, have provided solid evidence for the existence of an intrinsic, angiotensin-generating system in the pancreas. The tissue RAS has a potential role in finely regulating exocrine and endocrine functions of the pancreas such as ductal anion secretion and islet hormonal secretion. Some of these effects may be exerted via the markedly vasoconstrictive effects of RAS. Of particular interest in this context are the recent epidemiological data showing that administration of angiotensin-converting enzyme inhibitors appears to be protective against the development of diabetes in hypertensive patients. Moreover, the upregulation of pancreatic RAS has been shown to occur during chronic hypoxia. The significance of changes in pancreatic RAS could have a potential role in acute pancreatitis, islet transplantation and in different shock states, by causing a further decrease of blood perfusion in the pancreas. PMID- 11432371 TI - Twenty-year mortality in a 1981 cohort of homosexuals with gonorrhoea: a preliminary estimate. PMID- 11432372 TI - HIV screening in active tuberculosis patients in rural Gujurat, India. PMID- 11432373 TI - Does the tamoxifen increase thyroid dysfunction after loco-regional irradiation of breast cancer? PMID- 11432374 TI - [Sense and nonsense of information on temporomandibular disorders found on Internet]. PMID- 11432375 TI - Testing for detailed balance (microscopic reversibility in ion channel gating. PMID- 11432376 TI - Lymphoscintigraphy and lymphatic mapping for identification of sentinel lymph nodes. AB - The recently introduced technique of sentinel lymph node dissection (SLND) may replace complete axillary lymph node dissection for axillary staging of early breast cancer. Successful SLND is predicated on meticulous delineation of the lymphatic pathway and sentinel node(s). Currently employed lymphatic mapping materials include vital blue dyes and radioactive tracers. Techniques of intraoperative lymphatic mapping and SLND using dye, tracer, or both have high success rates in the hands of experienced investigators, but their routine and widespread use awaits resolution of questions about the timing, dose, and type of radioactive tracer; the optimal lymphatic mapping technique; indications and contraindications for SLND; and certification of qualified surgeons, pathologists, and nuclear medicine physicians. PMID- 11432377 TI - [Symposium "The dignity of animals--significance in the enrichment of human life". Basle, March 15-16, 2001]. PMID- 11432378 TI - [Animal experimentation and animal rights. Evangelical Academy Bad Boll, March 23 25, 2001]. PMID- 11432379 TI - [Cosmetics--European Parliament demands marketing prohibition]. PMID- 11432380 TI - [The 117th Internal Medicine Seminar (The 3rd Internal Medicine Seminar in Ostrava). Diabetes mellitus at the threshold of the 21st century. Ostrava, 30 November 2000]. PMID- 11432381 TI - Bitter pills, bad medicine. PMID- 11432382 TI - Strabology report of the 27th annual meeting of the Americans Association for Pediatric Ophthalmology and Strabismus. The Walt Disney World Swan Hotel, Orlando, Florida, March 21-25, 2001. PMID- 11432383 TI - Surgery for fully accommodative esotropia. PMID- 11432384 TI - Web alert. PMID- 11432385 TI - Vaccines, CJD including variant CJD. PMID- 11432386 TI - Pupil management during phacoemulsification in patients with iridoschisis. PMID- 11432387 TI - Popperian falsification of methods of assessing astigmatism. PMID- 11432388 TI - Ballooning of the conjunctiva during phacoemulsification. PMID- 11432389 TI - Gas sandwich technique for IOL support in combined cataract and vitreoretinal surgery. PMID- 11432390 TI - Delayed reappearance of a dislocated lens. PMID- 11432391 TI - Consultation section. Cataract surgery problem. PMID- 11432392 TI - [Charming buttocks]. PMID- 11432393 TI - Cardiomyocyte apoptosis contributes to the development of catacholamine cardiomyopathy. PMID- 11432394 TI - Intra-day variations in urinary pyrimidines in ornithine carbamoyltransferase deficiency and healthy individuals. PMID- 11432395 TI - Extracellular-superoxide dismutase in cerebrospinal fluid from infants/children. PMID- 11432396 TI - Fabry disease: enzymatic diagnosis in dried blood spots on filter paper. PMID- 11432397 TI - Endoscopic frontal branch neurectomy, corrugator myectomy, and brow lift for forehead asymmetry after facial nerve palsy. PMID- 11432398 TI - Expanded applications for transconjunctival lower lid blepharoplasty. PMID- 11432399 TI - Staged reconstruction after gunshot wounds to the abdomen. AB - Immediate closure of abdominal incisions after exploration and treatment of gunshot wounds is not always feasible or advisable. Significant bowel edema after massive fluid resuscitation might preclude primary closure, whereas any attempt to close under tension might result in complications ranging from wound dehiscence, infection, and necrosis to the abdominal compartment syndrome with abdominal, cardiopulmonary, and renal complications. For these difficult cases, the open technique has been recommended. The abdomen is left open and is closed when the patient's condition permits. When immediate wound approximation is not possible, temporary coverage can be achieved with a mesh, patch, or a split thickness skin graft and the definitive reconstruction is deferred for a more optimal time. The purpose of this retrospective study is to report the authors' experience with staged abdominal wall reconstruction after gunshot wounds. From 1989 to 1998, 1933 patients underwent exploratory laparotomy for penetrating wounds to the abdomen. Twenty-nine patients in grave condition and with multiple medical problems were comanaged by the Trauma and Plastic Surgery Services at Cook County Hospital with the following protocol: The abdomen was initially left open and exposed viscera were covered with a variety of methods, including a Gore Tex patch (W. L. Gore and Associates, Inc., Flagstaff, Ariz.). A split-thickness graft was subsequently placed on the granulation tissue over viscera at an average of 14 days after the last laparotomy. These planned ventral hernias were definitively treated at an average of 7 months after the skin grafting procedure, primarily using the components separation technique. In 24 patients, the fascia was closed primarily without tension, while five patients required the use of synthetic mesh to restore fascial continuity. Nine patients underwent closure of a colostomy or repair of fistulas simultaneously with abdominal wall reconstruction. One patient developed a postoperative hernia, two developed superficial wound dehiscence that healed without further surgery, and one required re-exploration for a failed anastomosis after colostomy closure. All but one patient maintained a stable abdominal wall after the reconstruction. The authors concluded that staged abdominal wall reconstruction should be primarily recommended for patients with complex abdominal wounds and a compromised general condition that precludes primary closure. With this treatment protocol, patients can recover faster from their trauma surgery and the risk of perioperative complications can be reduced. After final reconstruction, the continuity, stability, and strength of the abdominal wall are maintained in the vast majority of cases with the use of autogenous tissue and without the need for alloplastic material. With close cooperation between the trauma team and the plastic surgeon and appropriate timing and planning of each stage, the success rate of the technique is high and the incidence of complications limited. PMID- 11432400 TI - TH1/TH2,3 imbalance due to cytokine-producing NK, gammadelta T and NK-gammadelta T cells in murine pregnancy decidua in success or failure of pregnancy. AB - PROBLEM: Recurrent spontaneous abortion in DBA/2-mated CBA/J mice has been attributed to the production of Th1 cytokines (tumor necrosis factor [TNF]-alpha and interferon [IFN]-gamma) by asialoGM1+ natural killer (NK) cells and Vgamma1.1delta6.3+ T cells that infiltrate decidua by day 6.5, during the peri implantation period. Abortions can be prevented by a second population of Vgamma1.1delta6.3 cells, which infiltrate on day 8.5 of gestation, and produce the Th2 cytokine interleukin (IL)-10 and Th3 cytokine transforming growth factor (TGF)-beta2. In low abortion rate immunocompetent mice, most of the TGF-beta2 is derived from gammadelta T cells. However, TGF-beta2-producing cells are present in the decidua of pregnant severe combined immune deficient (SCID) mice, which lack gammadelta T cells. METHODS: The cells in day 13.5 decidua of CBA x DBA/2 matings and SCID x SCID matings were identified using flow cytometry and combined surface staining for gammadelta and/or asialoGM1, and intracellular cytokine staining for TNF-alpha, IFN-gamma, and TGF-beta2,3. RESULTS: TGF-beta2 and TNF alpha were found in asialoGM1+ NK cells in SCID mouse decidua. In CBA x DBA/2 mated mice, two major and one minor subsets of cytokine-positive cells were identified: -gammadelta-only T cells, double positive asialoGM1+ gammadelta+ (NK gammadelta T) cells, and a small number of asialoGM1 +gammadelta- NK-only cells. The NK-only and NK-gammadelta T subsets showed a greater Th1/Th2,3 pattern of intracellular staining compared with the gammadelta-only subset. In the CBA x DBA/2 and SCID x SCID systems, Th1/Th2,3 ratios could not predict actual observed abortion rates but did correlate with susceptibility to abortions (if exposed to an additional stimulus such as stress). The known effect of in vivo administration of anti-asialoGM1 antibody on abortion rates within groups of mice exposed to such stresses could also be predicted. CONCLUSION: gammadelta+ cells in decidua (e.g. Vgamma1+ cells which can recognize trophoblasts) differ based on the presence or absence of the NK marker-asialo-GM1. NK-gammadelta T cells may be quite important in the Th1 response in early pregnancy that predisposes to abortions in CBA x DBA/2 matings, whereas gammadelta T-only cells appear to be protective. In pregnant SCID mice, the TNF-alpha+/TGF-beta2+ NK population is greatly expanded. An activating stimulus (such as stress or endotoxin) appears to be as important in triggering abortions, as is the Th1/Th2,3 ratio at the feto maternal interface. PMID- 11432401 TI - Immunoregulatory activity, biochemistry, and phylogeny of ovine uterine serpin. AB - PROBLEM: During pregnancy, the endometrium of the ewe secretes a progesterone induced member of the serpin superfamily of serine proteinase inhibitors called ovine uterine serpin (OvUS) that has immunosuppressive properties. METHOD: Review of the literature. RESULTS AND CONCLUSIONS: OvUS inhibits a wide variety of immune responses, including mixed lymphocyte reaction, mitogen-stimulated lymphocyte proliferation, and T cell-dependent antibody production. Recent data have suggested that OvUS functions by inhibiting protein kinase C and interleukin 2-mediated events. OvUS and similar genes present in cattle and pigs diverged from other serpins prior to the divergence of artiodactyls. Since this time, the serpins have apparently undergone adaptive evolution that has led to a conformational state and biological functions distinct from prototypical serpins. Thus, it is likely that these proteins have an important role in the reproductive biology of Artiodactyla. Several lines of evidence suggest that, in sheep, OvUS functions to mediate the immunosuppressive effects of progesterone and prevent immunological rejection of the fetal allograft. PMID- 11432402 TI - Unique susceptibility of the fetal thymus to feline immunodeficiency virus infection: an animal model for HIV infection in utero. AB - PROBLEM: Human infants infected in utero with HIV develop thymus insufficiency and progress to AIDS sooner than infants infected peripartum. However, direct analysis of the thymus is difficult due to limited tissue access and variable timing of vertical transmission. METHOD OF STUDY: Fetal and neonatal cats were inoculated with feline immunodeficiency virus (FIV) at an equivalent infectious dose. The thymus, blood, and lymph nodes were harvested and compared at 23 and 46 days post-inoculation (p.i.) and also compared to sham-inoculated, age-matched controls. Lymphocyte phenotypes were analyzed by flow cytometry and virus burden was quantified in histologic sections and by virus isolation from plasma. RESULTS: Fetal cats inoculated with FIV had acute thymus atrophy at birth, which coincided with peak viremia. At 46 days p.i., thymus size and cell composition rebounded and supported increased productive infection. In contrast, neonatal cats inoculated with FIV developed chronic thymus atrophy and degeneration, which was associated with decreasing productive infection and low-level viremia. CONCLUSIONS: The fetal thymus is uniquely vulnerable to acute, transient depletion and high-level productive infection. The neonatal thymus is less vulnerable to acute changes, and responds through progressive atrophy and declining productive infection. Reduced immune competence, as reflected by the failure to control virus replication, may contribute to the accelerated progression of FIV and HIV infections in utero. PMID- 11432403 TI - Asymmetric antibodies and pregnancy. AB - PROBLEM: Asymmetric IgG antibodies (AAb) possess a mannose-rich oligosaccharide residue bound to one of the Fab regions, making them unable to activate immunoeffector mechanisms. The proportion of asymmetric antibodies is increased after prolonged immunization with particulate antigens like cellular spleen cells. During pregnancy, AAb were found in serum and bound to placenta with specific activity to paternal antigens. No previous reports about the status of AAb in recurrent spontaneous abortion (RSA) patients have been published to date. Therefore, the aim of the present study was to analyze the percentage of asymmetric IgG molecules in serum samples of (a) healthy pregnant and non pregnant women, (b) pregnant women with a history of RSA, and (c) non-pregnant RSA patients receiving paternal lymphocyte immunotherapy (LIT) or intravenous gammaglobulin therapy (IVIgs). METHOD OF STUDY: A previously-described differential ELISA technique was used to determine the percentage of IgG that was of the asymmetric type. RESULTS: During normal pregnancy, there was an increase in the percentage of high ConA affinity IgG serum molecules with a major increase at the second trimester. Pregnant RSA patients at the second trimester had lower values. When evaluating non-pregnant RSA patients who received LIT, it was observed that the immunized patients expressed a higher percentage of asymmetric IgG antibodies. The pregnant patients who received IVIgs had a percentage of AAbs comparable to normal pregnant patients. Additionally, the presence of IgG asymmetric molecules was confirmed in commercial gammaglobulin preparations. CONCLUSION: Results suggest a protective role of AAb during pregnancy. PMID- 11432404 TI - Characterization of NKT cells in human peripheral blood and decidual lymphocytes. AB - PROBLEM: To examine whether natural killer (NKT) cells are present in human pregnancy decidua. METHOD OF STUDY: We calculated the percentage of CD3+CD161+Valpha 24+-NKT cells in peripheral blood and early pregnancy decidua, and analyzed intracellular cytokines, interleukin (IL)-4 and interferon (IFN)gamma in NKT cells using flow cytometry. RESULTS: A distinct subset of CD3+ CD161+ lymphocytes expressing an invariant antigen receptor encoded by the Valpha24 and Vbeta11 segment was accumulated in the decidua. In pregnant subjects the percentages of NKT cells were significantly increased in the decidua compared with peripheral blood. Both NKT cells in the decidua and the peripheral blood had an ability to rapidly produce cytokine associated with Th1 (IFNgamma) and Th2 (IL 4). Interestingly, the percentages of IL-4 and IFNgamma producing NKT cells were significantly higher in the decidua compared with the peripheral blood. CONCLUSIONS: These findings suggest that NKT cells might control the Th1/Th2 balance by producing IL-4 and IFNgamma at the feto-maternal interface. PMID- 11432405 TI - Murine stress-triggered abortion is mediated by increase of CD8+ TNF-alpha+ decidual cells via substance P. AB - PROBLEM: Stress is known to induce abortions in mice and humans. Increased levels of abortogenic type 1 helper T-cell cytokines and decreased levels of pregnancy protective cytokines could be linked to stress-triggered embryonic loss. Stress promotes neurotransmitter substance P (SP) release in tissues. SP increases the production of decidual tumor necrosis factor (TNF)-alpha, whereby the phenotype of these TNF-alpha-producing cells is hypothetical. The objective of the present study was to identify decidual TNF-alpha-producing cell populations that are involved in stress-induced murine abortion. METHOD: DBA/2J-mated CBA/J female mice were exposed to ultrasonic sound stress on day 5.5 of pregnancy. The mice were randomized and half were treated with the SP NK1-receptor antagonist (SP-RA) RP 67580 (200 microg/mouse). Frequency and cytokine profile of CD8+ cells were evaluated by immunohistochemistry and flow cytometry. Degranulation of uterine mast cells was examined histologically. RESULTS: On day 13.5 of pregnancy, the uteri were removed and the resorption rate was calculated. A mean resorption rate of 38.4% was detected in stressed mice (n = 10) compared to 13.1% in non-stressed control mice (n = 11, P < 0.01). Injection of SP-RA decreased the abortion rate to 18.4% in stressed mice (n = 19, P < 0.01). Flow cytometry revealed a stress related increase of TNF-alpha+/CD8+ decidual T cells, which could be abrogated by SP-RA (P < 0.05). No significant differences could be observed in numbers of mast cells and total CD8+ cells in situ. CONCLUSION: Our data suggest that stress triggered abortion is mediated by SP, and SP receptor blockade abrogates stress triggered abortion via reduced production of TNF-alpha by CD8+ T cells. PMID- 11432406 TI - Bovine trichomoniasis as a model for development of vaccines against sexually transmitted disease. AB - PROBLEM: Human sexually transmitted diseases (STDs) are widespread but effective vaccines are rare. Experimental and commercially available vaccines for bovine trichomoniasis have been well studied. Principles for immune protection of the female genital tract derived from studies of bovine trichomoniasis may be generally applicable to human trichomoniasis and other STDs. METHOD OF STUDY: A bovine model of trichomoniasis has been developed for study of mechanisms of immunoprophylaxis. RESULTS: Both systemic and local immunization with an immunoaffinity purified antigen cleared the genital tract of trichomonads significantly earlier than non-immunized controls. Predominantly IgA responses or predominantly IgG responses in uterine and vaginal secretions were essentially equally protective. Uterine and vaginal IgA responses could be induced by systemic priming and local boosting via either the vaginal or nasal mucosa. In either case, lymphoid aggregates were formed in the uterine and vaginal mucosa which were not present in the genital mucosa of naive animals. CONCLUSIONS: Systemic immunization or systemic priming with local boosting protects against bovine trichomoniasis via IgG or IgA antibodies (respectively) to a major surface antigen of trichomonads. Immunization of the genital mucosa results in formation of inductive sites for a local IgA response. PMID- 11432407 TI - Author index to volume 86, 2001. PMID- 11432408 TI - Groin hernia surgery: a systematic review. AB - BACKGROUND: An extensive volume of clinical research has been undertaken on the use of surgery for groin hernias. For many years there has been a large number of different methods of repairing hernias and, with the introduction of laparoscopic surgery, this has increased further. It is uncertain which method is the best in terms of safety and effectiveness. OBJECTIVES: This review was undertaken to compare the outcomes following different surgical procedures to treat groin hernias in adults. It sought answers to six questions: 1)Which method of surgery (including open procedures and laparoscopic surgery) is the safest and most effective for inguinal hernia repair? 2) Is local anaesthesia a safe and effective alternative to general anaesthesia? 3) Is there a difference in outcome between specialist and non-specialist surgeons? 4) Is day-case as safe and effective as inpatient surgery? 5) Is synchronous bilateral hernia repair as safe and effective as delayed repair? 6) Which method of surgery is the safest and most effective for femoral hernia repair? METHODS: The primary measure of effectiveness used was the proportion of hernia repairs in which there was a recurrence. Secondary outcome measures included complications, post-operative pain, wound infection, time to return to normal activities and/or return to work. A systematic search of the literature (up to February 1996) was undertaken using a variety of approaches. the methodological quality of all prospective comparative studies (45 randomised trials and 26 non-randomised trials/prospective cohort studies) was assessed using a standard checklist. RESULTS: Some of the variation in findings from different studies is likely to be due to methodological differences rather than differences in the effectiveness of the surgical procedures. The main methodological shortcomings of the studies that have been performed are: lack of agreed method for assessing severity of hernias; failure to take confounding into account in non-randomised studies; variation in length of follow-up; poor external validity; lack of objective measures of outcome; and inadequate statistical power. These problems severely limit the conclusions that can be drawn from the literature. PMID- 11432409 TI - A fast and easy screening method for voice disorders among teacher students. AB - A recent study concerning voice disorders among future teachers showed that about 20% of the students had a voice disorder. Most of the disorders were organic. In the current study, we describe a voice-screening method that can be administered by the health care personnel in order to select students for further examination by the speech language therapist and/or phoniatrician. The screening method consists of a perceptual assessment of voice quality and a questionnaire concerning vocal symptoms. As criterion for further medical investigation and voice therapy, we selected a score of 35 mm or above on a visual analogue scale assessing Grade, i.e. overall grade of hoarseness and/or two or more weekly or more often occurring vocal symptoms. The results showed that health care personnel with some training in assessing voices using the questionnaire are competent to perform a rough voice screening on students. PMID- 11432410 TI - The impact of impaired vocal quality on children's ability to process spoken language. AB - This paper investigated the effect of voice quality on children's ability to process spoken language. A group of 24 children, mean age 11 years 5 months, listened to a series of recorded short passages, half spoken by a female with normal voice and half spoken by a female with a classic vocal impairment (dysphonic voice). The children were tested for their ability to recall words and to draw a final target inference. Children performed better on both preceding indices when listening to the normal voice. The implications of the findings are discussed, with particular reference to the classroom situation. PMID- 11432411 TI - Levels of cognitive and linguistic development in Angelman syndrome: a study of 20 children. AB - Angelman syndrome (AS) is a genetic disorder associated with severe developmental delay. The purpose of this study was to investigate cognitive and linguistic development in AS. Piaget's developmental model was used to evaluate the test results. The participants comprised 20 children (14 boys and 6 girls) aged 2-14 years (median age 7.4 years). AS was diagnosed either according to typical clinical criteria or confirmatory genetic testing. Cognitive functioning was evaluated with Griffiths' Mental Development Scale. Language development was also evaluated with Receptive-Expressive Emergent Language Scale 2 (REEL-2). Cognitive functioning, based on results on the Performance Scale, never exceeded Piaget's sensorimotor stage, 0-2 years. The median mental age for language development was 9 months. Expressive verbal vocabulary consisted of less than 2 words (n = 11), 2 3 words (n = 7) and 4-5 words (n = 2). Analyses according to REEL-2 did not indicate a consistent discrepancy between impressive and expressive language. PMID- 11432412 TI - Voice source differences between falsetto and modal registers in counter tenors, tenors and baritones. AB - Vocal registers are generally assumed to be associated with the voice source, i.e. the pulsating transglottal airflow. The waveform of this airflow was analysed by inverse filtering in professional singers, four counter tenors, five tenors, and four baritones singing the syllable [pae:] in soft, middle, and loud voice in modal and falsetto/counter tenor register. Subglottal pressure, estimated from the intra-oral pressure during the occlusion for the consonant [p], closed quotient, relative glottal leakage, and the relative level of the fundamental were analysed. The counter tenors used comparatively low subglottal pressures and mostly showed a closed phase in their flow glottogram waveform. For a given value of the closed quotient, the fundamental tended to be stronger in falsetto than in modal register. The observed voice source differences between the registers seem related to a greater vocal fold thickness in modal than in falsetto register. PMID- 11432413 TI - Review: occupational risks for voice problems. AB - The purpose of this paper is to provide a cohesive review of the literature regarding the functional consequences of voice problems and occupational risk factors for them. The salient points are as follows. According to conservative estimates, approximately 28,000,000 workers in the US experience daily voice problems. Many people who experience voice problems perceive them to have a negative impact on their work and their quality of life. Estimates based on empirical data suggest that, considering only lost work days and treatment expenses, the societal cost of voice problems in teachers alone may be of the order of about $2.5 billion annually in the US. In fact, across several countries, "teacher" consistently emerges as the common occupation most likely to seek otorhinolaryngological (ORL) evaluation for a voice problem. Other occupational categories likely to seek ORL examination for a voice problem are singer, counselor/social worker, lawyer, and clergy. Finally, US Census data indicate that keyboard operators may be at special risk for voice problems because of a near-epidemic growth of repetitive strain injury (RSI), which can adversely affect the voice especially when speech recognition software is implemented. This paper discusses frequency data, quality of life data, and treatment considerations for these voice-related occupational issues. PMID- 11432414 TI - Use of antimicrobials in veterinary medicine and mechanisms of resistance. AB - This review deals with the application of antimicrobial agents in veterinary medicine and food animal production and the possible consequences arising from the widespread and multipurpose use of antimicrobials. The various mechanisms that bacteria have developed to escape the inhibitory effects of the antimicrobials most frequently used in the veterinary field are reported in detail. Resistance of bacteria to tetracyclines, macrolide-lincosamide streptogramin antibiotics, beta-lactam antibiotics, aminoglycosides, sulfonamides, trimethoprim, fluoroquinolones and chloramphenicol/florfenicol is described with regard to enzymatic inactivation, decreased intracellular drug accumulation and modification/protection/replacement of the target sites. In addition, basic information is given about mobile genetic elements which carry the respective resistance genes, such as plasmids, transposons, and gene cassettes/integrons, and their ways of spreading via conjugation, mobilisation, transduction, and transformation. PMID- 11432415 TI - Resistance to antibiotics in the normal flora of animals. AB - The normal bacterial flora contains antibiotic resistance genes to various degrees, even in individuals with no history of exposure to commercially prepared antibiotics. Several factors seem to increase the number of antibiotic-resistant bacteria in feces. One important factor is the exposure of the intestinal flora to antibacterial drugs. Antibiotics used as feed additives seem to play an important role in the development of antibiotic resistance in normal flora bacteria. The use of avoparcin as a feed additive has demonstrated that an antibiotic considered "safe" is responsible for increased levels of antibiotic resistance in the normal flora enterococci of animals fed with avoparcin and possibly in humans consuming products from these animals. However, other factors like stress from temperature, crowding, and management also seem to contribute to the occurrence of antibiotic resistance in normal flora bacteria. The normal flora of animals has been studied with respect to the development of antibiotic resistance over four decades, but there are few studies with the intestinal flora as the main focus. The results of earlier studies are valuable when focused against the recent understanding of mobile genetics responsible for bacterial antibiotic resistance. New studies should be undertaken to assess whether the development of antibiotic resistance in the normal flora is directly linked to the dramatic increase in antibiotic resistance of bacterial pathogens. Bacteria of the normal flora, often disregarded scientifically, should be studied with the intention of using them as active protection against infectious diseases and thereby contributing to the overall reduction of use of antibioties in both animals and humans. PMID- 11432416 TI - Importance of integrons in the diffusion of resistance. AB - Horizontal transfer of resistance genes is a successful mechanism for the transmission and dissemination of multiple drug resistance among bacterial pathogens. The impact of horizontally transmitted genetic determinants in the evolution of resistance is particularly evident when resistance genes are physically associated in clusters and transferred en bloc to the recipient cell. Recent advances in the molecular characterisation of antibiotic resistance mechanisms have highlighted the existence of genetic structures. called integrons, involved in the acquisition of resistance genes. These DNA elements have frequently been reported in multi-drug resistant strains isolated from animals and humans, and are located either on the bacterial chromosome or on broad-host-range plasmids. The role of integrons in the development of multiple resistance relies on their unique capacity to cluster and express drug resistance genes. Moreover, the spread of resistance genes among different replicons and their exchange between plasmid and bacterial chromosome are facilitated by the integration of integrons into transposable elements. The association of a highly efficient gene capture and expression system, together with the capacity for vertical and horizontal transmission of resistance genes represents a powerful weapon used by bacteria to combat the assault of antibiotics. PMID- 11432417 TI - Resistance to trimethoprim and sulfonamides. AB - Sulfonamides and trimethoprim have been used for many decades as efficient and inexpensive antibacterial agents for animals and man. Resistance to both has, however, spread extensively and rapidly. This is mainly due to the horizontal spread of resistance genes, expressing drug-insensitive variants of the target enzymes dihydropteroate synthase and dihydrofolate reductase, for sulfonamide and trimethoprim, respectively. Two genes, sul1 and sul2, mediated by transposons and plasmids, and expressing dihydropteroate synthases highly resistant to sulfonamide, have been found. For trimethoprim, almost twenty phylogenetically different resistance genes, expressing druginsensitive dihydrofolate reductases have been characterized. They are efficiently spread as cassettes in integrons, and on transposons and plasmids. One particular gene, dfr9, seems to have originally been selected in the intestine of swine, where it was found in Escherichia coli, on large plasmids in a disabled transposon, Tn5393, originally found in the plant pathogen Erwinia amylovora. There are also many examples of chromosomal resistance to sulfonamides and trimethoprim, with different degrees of complexity, from simple base changes in the target genes to transformational and recombinational exchanges of whole genes or parts of genes, forming mosaic gene patterns. Furthermore, the trade-off, seen in laboratory experiments selecting resistance mutants, showing drug-resistant but also less efficient (increased Kms) target enzymes, seems to be adjusted for by compensatory mutations in clinically isolated drug-resistant pathogens. This means that susceptibility will not return after suspending the use of sulfonamide and trimethoprim. PMID- 11432418 TI - Quinolone resistance in Escherichia coli. AB - Escherichia coli is an important pathogen of animals and humans that causes great financial cost in food production by causing disease in food animals. The quinolones are a class of synthetic antimicrobial agents with excellent activity against Escherichia coli and other Gram-negative bacteria used in human and veterinary medicine. Different quinolones are used to treat various conditions in animals in different parts of the world. All members of this class of drug have the same mode of action: inhibition of topoisomerase enzymes, DNA Gyrase and Topoisomerase IV. Escherichia coli can become resistant to quinolones by altering the target enzymes, reducing permeability of the cell to inhibit their entry, or by actively pumping the drug out of the cell. All these resistance mechanisms can play a role in high-level fluoroquinolone resistance, however target site mutations appear to be most important. As all quinolones act in the same way resistance to one member of the class will also confer decreased susceptibility to all members of the family. Quinolone resistant Escherichia coli in animals have increased in numbers after quinolone introduction in a number of different case studies. The resistance mechanisms in these isolates are the same as those in resistant strains found in humans. Care needs to be taken to ensure that quinolones are used sparingly and appropriately as highly resistant strains of Escherichia coli can be selected and may pass into the food chain. As these drugs are of major therapeutic importance in human medicine, this is a public health concern. More information as to the numbers of quinolone resistant Escherichia coli and the relationship between resistance and quinolone use is needed to allow us to make better informed decisions about when and when not to use quinolones in the treatment of animals. PMID- 11432419 TI - Epidemiology of resistance to quinolones in Salmonella. AB - Fluoroquinolones account for about 11% of antimicrobial prescriptions in human medicine worldwide and represent the drug of choice for the treatment of a wide range of human infectious diseases. They were introduced into veterinary medicine in Europe in the late 1980s and early 1990s and in the USA in 1995. Following their introduction, resistant strains of bacteria, including Salmonella, started to emerge. Resistance to quinolones depends on chromosomal mutations and the subsequent spread of resistant clones. While the selective pressure caused by the use of quinolones facilitates their epidemic transmission, the resistant mutants may spread independently of quinolone use. In view of the key role of this group of antimicrobials in human medicine and the position of Salmonella as the leading cause of food-borne infections in many countries, the public health hazard posed by quinolone-resistant zoonotic Salmonella serovars has been a subject of concern. The fluoroquinolones are on the WHO list of drugs that should be reserved for human use. Considering the mounting evidence that quinolone resistant zoonotic Salmonella are the cause of severe, sometimes fatal, infections in humans, the use of fluoroquinolones in food animals should be discontinued or severely restricted. Such an intervention should be accompanied by prudent use measures involving all other groups of antimicrobials to reduce the need for fluoroquinolones in veterinary medicine. PMID- 11432420 TI - Mechanisms of quinolone resistance in Salmonella. AB - As in other Gram-negative bacteria, mechanisms of resistance to quinolones in Salmonella include target gene mutations, active efflux, and decreased outer membrane permeability. However, the exact contribution of these individual mechanisms to resistance, which may nevertheless interplay to reach high-level resistance, has not yet clearly been defined as in other bacteria such as Escherichia coli. This paper reviews the current state of knowledge of quinolone resistance mechanisms in Salmonella by comparison with that of E. coli and future directions of research with particular attention to the recent development of efflux pump inhibitors as possible means of avoiding the emergence and spread of fluoroquinolone resistance. PMID- 11432421 TI - Molecular characterization, spread and evolution of multidrug resistance in Salmonella enterica typhimurium DT104. AB - Multidrug-resistant Salmonella enterica serovar Typhimurium phage type DT104 has emerged during the last decade as a global health problem because of its involvement in diseases in animals and humans. Multidrug-resistant DT104 strains are mostly resistant to ampicillin, chloramphenicol, streptomycin, sulfonamides and tetracyclines (ACSSuT resistance type). The genes coding for such resistances are clustered on the chromosome. This paper reviews new developments in the characterization of S. enterica Typhimurium DT104, its chromosomal antibiotic resistance genes and their spread among other S. enterica Typhimurium phage types and other S. enterica serovars, the development of specific detection methods, virulence characteristics, and the evolution of multidrug-resistance with regard to the emergence of quinolone resistance. PMID- 11432422 TI - Antimicrobial resistance of thermophilic Campylobacter. AB - Campylobacter has become the leading cause of zoonotic enteric infections in developed and developing countries world-wide. Antimicrobial resistance has emerged among Campylobacter mainly as a consequence of the use of antimicrobial agents in food animal production. Resistance to drugs of choice for the treatment of infections, macrolides and fluoroquinolones has emerged as a clinical problem and interventions to reduce this are recommended. Resistance to fluoroquinolones and macrolides is mediated by chromosomal mutations. Resistance to other relevant antimicrobial agents, mediated by acquired resistance genes, has not become widespread so far. However, resistance genes originating from both Gram-positive and Gram-negative bacterial species have been found, showing the potential for acquired resistance to emerge in Campylobacter. PMID- 11432423 TI - Antimicrobial resistance in pasteurella and mannheimia: epidemiology and genetic basis. AB - Isolates of the genera Pasteurella and Mannheimia cause a wide variety of diseases of great economic importance in poultry, pigs, cattle and rabbits. Antimicrobial agents represent the most powerful tools to control such infections. However, increasing rates of antimicrobial resistance may dramatically reduce the efficacy of the antimicrobial agents used to control Pasteurella and Mannheimia infections. This review presents a short summary of the infections caused by Pasteurella and Mannheimia isolates in food-producing animals and the possibilities of preventing and controlling primary and secondary pasteurellosis. Particular reference is given to antimicrobial chemotherapy and the resistance properties of Pasterurella and Mannheimia isolates. The genetic basis of the most predominant resistance properties such as resistance to beta lactam antibiotics, tetracyclines, aminoglycosides, sulfonamides, and chloramphenicol is discussed. This is depicted with reference to the role of plasmids and transposons in the spread of the resistance genes among Pasteurellaceae and members of other bacterial families and genera. PMID- 11432424 TI - Antimicrobial resistance in staphylococci from animals with particular reference to bovine Staphylococcus aureus, porcine Staphylococcus hyicus, and canine Staphylococcus intermedius. AB - Besides their role as commensals on the skin and mucosal surfaces, staphylococci may be involved in a wide variety of diseases in animals. Staphylococcal infections in animals are mainly treated with antimicrobial agents and as a consequence, staphylococci from animal sources have developed and/or acquired resistance to the respective antimicrobial agents. Resistance statistics obtained from national monitoring programmes on staphylococci from cattle and pigs, but also from surveillance studies on staphylococci involved in diseases in dogs are reported and reviewed with regard to their comparability. This review mainly focusses on the genetic basis of antimicrobial resistance in staphylococci of animal origin. Particular attention is paid to resistance to those antimicrobial agents which are most frequently used in veterinary medicine, but also to antimicrobial agents, such as chloramphenicol and mupirocin, which are used in specific cases for the control of staphylococcal infections in pets and companion animals. In addition, plasmids and transposons associated with the respective resistance properties and their ways of spreading between members of the same or different staphylococcal species, but also between staphylococci and other gram positive bacteria, are described. PMID- 11432425 TI - Molecular tools for the characterisation of antibiotic-resistant bacteria. AB - This review will discuss a number of molecular tools which are currently used as well as some innovative approaches for the characterisation of antibiotic resistant bacterial strains. Various methods involved in the detection and characterisation of genes and mutations associated with antibiotic resistance and that are used for strain typing as part of epidemiological studies, are described. Furthermore, a few examples are discussed in which the results of both gene and strain characterisation are combined to investigate the underlying mechanism of the spread of antibiotic resistance. Some of the available molecular techniques are heavily supported by the existence of databases on the Internet. These databases either contain a fast growing amount of sequence information or a large number of allelic or fingerprint profiles. The current progress in applied DNA technology and the ongoing projects on the elucidation of the whole genomic sequence of bacterial species have lead and will further lead to the development and application of sophisticated new strategies for the analysis of antibiotic resistant bacterial strains. PMID- 11432426 TI - New trends in regulatory rules and surveillance of antimicrobial resistance in bacteria of animal origin. AB - Since the introduction in the 1940s of antibiotics as drugs against bacterial infections in human and then veterinary medicine, two major events have caused a shift in the antibiotherapy era: (1) the emergence of resistant bacteria and (2) the awareness of the limits of new drug development. It rapidly became urgent to set up measures in order to evaluate the importance of resistant bacteria and their origin as well as to limit the dissemination of resistant vectors (bacteria and bacterial genes). This led to the establishment of guidelines and regulatory rules necessary for risk assessment and clearly dependent upon monitoring and research organisations. At a veterinary level, the possible dissemination of multiresistant bacteria from animals to humans, through feeding, urged various national European and international institutions to give general recommendations to monitor and contain the emergence and diffusion of resistant strains. This paper gives an overview of the evolution of regulatory rules and monitoring systems dealing with multiresistant bacteria. PMID- 11432427 TI - Anaesthetic management of baboons undergoing heterotopic porcine cardiac xenotransplantation. AB - A detailed anaesthetic technique for baboons (Papio anubis) undergoing heterotopic abdominal cardiac xenotransplantation is described. Twenty-two baboons served as transplant recipients. Donors were either crossbred farm pigs (Sus scrofa) (n = 4) or transgenic pigs (Sus scroefa) (n = 18) expressing human complement regulatory proteins on the endothelium. Intra-operative management was complicated by the physiological consequences of infrarenal. abdominal aortic cross-clamping, in addition to the immunological sequelae related to cross species transplantation. In choosing anaesthetics for this procedure, we considered the need for maximal cardiac stability throughout a long surgical procedure that required abdominal aortic cross-clamping to facilitate the implantation of an oversized porcine cardiac graft. Baboons received a balanced anaesthetic consisting of inhaled isoflurane in oxygen, intravenous fentanyl and intravenous pancuronium. The pharmacological techniques employed were found to be safe and reliable and were well tolerated by our recipients without any significant side-effects. PMID- 11432428 TI - Comparison of the value of pulsed-field gel electrophoresis, random amplified polymorphic DNA and amplified rDNA restriction analysis for subtyping Taylorella equigenitalis. AB - Eight strains of Taylorella equigenitalis were identified by a polymerase chain reaction using a primer pair specific to the 16S rDNA of T equigenitalis. These eight strains were chosen because they had previously been shown to represent eight distinct genotypes by pulsed-field gel electrophoresis analysis after separate digestion of the genomic DNA with ApaI or NotI. The eight strains could be classified into six or seven types by random amplified polymorphic DNA analysis using different kinds of primers. Amplified rDNA restriction analysis after separate digestion with five restriction enzymes, including AluI and MboI, of the 1,500 bp fragments of rDNA amplified by polymerase chain reaction did not discriminate the genomic variations among the eight strains of T equigenitalis. Thus, pulsed-field gel electrophoresis was shown to discriminate these eight organisms better than random amplified polymorphic DNA analysis, while amplified rDNA restriction analysis was found to be unsuitable for subtyping T equigenitalis. PMID- 11432429 TI - Measurement of faecal cortisol metabolites in cats and dogs: a non-invasive method for evaluating adrenocortical function. AB - The aim of this comparative study was to gain more information about the metabolism and excretion of glucocorticoids in cats and dogs in order to establish non-invasive methods for evaluating stressful conditions. Therefore, in a first experiment, [14C]cortisol was administered intravenously to 8 animals (two of each sex and species). Over a period of 6 days, faeces and urine were collected immediately after spontaneous defecation and urination. Marked species differences were found, as cats mainly excreted cortisol in the faeces (82% +/- 4% of the total recovered radioactivity), whereas in dogs only a small portion was found there (23% +/- 4%). The highest urinary radioactivity was observed after 9 +/- 3 h in cats and 3 +/- 1 h in dogs. Peak concentrations in the faeces occurred after 22 +/- 6 h in cats and after 24 +/- 4 h in dogs. Most of the radioactivity was not extractable with diethyl ether, indicating that the metabolites excreted in urine and faeces were mainly of the conjugated or polar unconjugated types. This was confirmed by RP-HPLC, which also revealed marked differences between cats and dogs concerning the metabolites formed. In addition, the immunoreactivity of the metabolites was tested in cortisol, corticosterone and 11-oxoaetiocholanolone EIAs. The latter, measuring 11,17-dioxoandrostanes (11,17-DOA) detected the highest quantities of immunoreactive metabolites in cats, but not in dogs. In a second experiment, the adrenal cortex of both species was stimulated by ACTH and, three weeks later, suppressed by dexamethasone. In this study, only faeces were collected over a period of 7 days. In both species, inter-animal variability in the basal and maximal/minimal faecal cortisol metabolite concentrations and the time course was observed. The 11 oxoaetiocholanolone EIA in cats and the cortisol EIA in dogs proved best suited for monitoring changes in adrenocortical activity. ACTH injections resulted in an increase above baseline values of 355%, (median) in 11,17-DOA concentrations in cats and of 702% in the concentrations of cortisol equivalents in dogs by about 25 h and 22 h (median) after injection, respectively. Minimal concentrations after dexamethasone administration were about 17% in cats and 31% in dogs (in relation to baseline values) and were reached in 66 h and 72 h, respectively. It was concluded that measuring cortisol metabolites in faeces should be a useful non-invasive tool for monitoring stress in carnivores. PMID- 11432430 TI - Studies on some blood parameters of crossbred calves with experimental Theileria annulata infections. AB - Subcutaneous inoculation of 1 ml of ground Theileria annulata tick tissue stabilate (0.75 tick equivalent) into crossbred calves (n = 6, average age 53 days) resulted in the development of acute theileriosis. The percentage parasitaemia was 71.7% +/- 3.3% on day 20 after inoculation. Macroschizonts were observed in lymphocytes and monocytes. Phagocytosed schizonts were observed in neutrophils, along with cytoplasmic vacuolation in monocytes and neutrophils. There was progressive decrease (p < 0.05) in the haemoglobin and packed cell volume, along with a marked reticulocytosis. Serum analysis revealed a decrease (p <0.05) in the concentrations of calcium, cholesterol and triglycerides, while there was an increase (p < 0.05) in the concentrations of blood urea nitrogen as compared to day 0 values. The total serum proteins, albumin and serum immunoglobulin concentrations and the albumin-to-immunoglobulin ratio showed marked decreases (p<0.05). Coagulopathies included thrombocytopenia and an increased prothrombin time, along with a non-significant increase in the bleeding time and activated partial thromboplastin time during the terminal stages of the disease. There was an increase in the osmotic fragility of erythrocytes during the disease. Morphological alterations in the erythrocytes were observed with the developing parasitaemia. PMID- 11432431 TI - Intramuscular kinetics and dosage regimens for pralidoxime in buffalo calves (Bubalus bubalis). AB - The plasma levels, disposition kinetics and a dosage regimen for pralidoxime (2 PAM) were investigated in male buffalo calves following single intramuscular administration (15 or 30 mg/kg). The effects of 2-PAM on various blood enzymes were also determined. The absorption half-life, elimination half-life, apparent volume of distribution and total body clearance of 2-PAM were 1.08 +/- 0.19 h, 3.14-3.19 h, 0.83-1.01 L/kg and 184.9-252.1 ml/(kg h), respectively. At doses of 15 and 30 mg/kg body weight, a plasma concentration > or = 4 microg/ml was maintained for up to 4 and 6 h, respectively. Pralidoxime significantly lowered the serum level of transferases, phosphatases and lactate dehydrogenase but did not influence the acetylcholinesterase and carboxylesterase enzymes. The most appropriate dosage regimen for 2-PAM in the treatment of organophosphate toxicity in buffaloes would be 25 mg/kg followed by 22 mg/kg at 8 h intervals. PMID- 11432432 TI - Comparison of immunoperoxidase plaque staining and neutralizing tests for canine distemper virus. AB - The use of immunoperoxidase plaque staining (IP) in the antibody test for canine distemper virus (CDV) and its diagnostic value were investigated. Antibody titres by the IP test and neutralizing test (NT) against CDV were compared in 114 specific pathogen-free (SPF) dogs, including 84 CDV-vaccinated and 30 unimmunized dogs. All of the unimmunized dogs, which had negative NT titres, also showed negative IP titres, and all of the CDV-vaccinated dogs, which had positive NT titres, showed positive IP titres. Furthermore, the IP titres showed similar changes with time after vaccination to the NT titres in five CDV-vaccinated SPF dogs. These findings indicate that the IP test has similar specificity to the NT. Next, the IP and NT titres were compared in 190 clinically suspected cases of canine distemper (CD), including 107 dogs with neurological signs and 83 dogs without neurological signs, and also in 50 clinically healthy dogs. Among the suspected cases of CD, of 46 dogs with neurological signs and 29 dogs without neurological signs, and which had negative NT titres, 33 (72%) and 19 (66%), respectively, showed positive IP titres. Many suspected cases of CD showed a high IP titre despite a low NT titre. In contrast, among healthy dogs, none of 9 NT negative dogs showed positive IP titres. The difference in the IP and NT titres (log10) [reciprocal of IP titre] - log10) [reciprocal of NT titre]) was measured to investigate the agreement between IP test and NT. The differences were 1.488 +/- 0.867, 1.245 +/- 0.833, 0.595 +/- 0.351 and 0.594 +/- 0.372, respectively, in the suspected cases of CD with neurological signs, the suspected cases of CD without neurological signs, the healthy dogs and the SPF dogs. The differences in the suspected cases of CD were significantly high compared to those for the SPF dogs (p < 0.001). Furthermore, in 8 dogs diagnosed as cases of CD from their paired sera, whose initial sera showed a markedly low NT titre compared to the IP titre, 5 dogs without neurological signs developed neurological signs later. These findings suggest that the IP test can detect CD during its early phases and in dogs with low antibody levels, undetectable by NT, and so would be useful for serological diagnosis. PMID- 11432433 TI - Antibody testing against canine coronavirus by immunoperoxidase plaque staining. AB - The application of the immunoperoxidase (IP) plaque staining procedure (IP test) to the diagnosis of canine coronavirus (CCV) infection was investigated. The IP test did not react with sera from either 15 specific pathogen-free (SPF) dogs or 7 SPF dogs immunized with a multivalent vaccine, including canine parvovirus type 2, canine distemper virus, canine adenovirus type 2, and canine parainfluenza virus. To compare the IP test with the neutralizing test (NT), sera from 240 healthy dogs and from 3 experimentally CCV-infected dogs were examined. All 60 sera positive for NT antibody were positive for IP antibody, and all 180 sera negative for NT antibody were negative for IP antibody in the healthy dogs. The IP titres showed similar changes with time after CCV inoculation to those of the NT titres in the experimentally infected dogs. These findings indicate that the IP test specifically detected anti-CCV antibodies. When the IP test and NT were compared in dogs with diarrhoeic signs. 2.1% of 48 sera and 20.3% of 74 sera, which were all negative for NT antibody, were positive for IP antibody in the dogs of under one year of age and at least one year of age, respectively. The difference between the IP and NT titres (log10 [reciprocal of IP titre] log10 [reciprocal of NT titre]) for the diarrhoeic dogs of under one year of age (2.350 +/- 0.931) was significantly larger than that for the healthy dogs (0.982 +/- 0.447) (p<0.0001), the NT titre being negative or very low, despite a high IP titre in many diarrhoeic dogs. Hence, the IP test is more able to detect anti-CCV antibodies, especially in dogs showing clinical signs. The IP-positivity rate was significantly higher in the diarrhoeic dogs of under one year of age (48.7%) than in the healthy dogs (25.0%) (chi2 = 19.844, p<0.0001), suggesting that CCV may contribute to diarrhoea in many juvenile dogs. PMID- 11432434 TI - Correlations to predict droplet size in ultrasonic atomisation. AB - In conventional two fluid nozzles, the high velocity air imparts its energy to the liquid and disrupts the liquid sheet into droplets. If the energy for liquid sheet fragmentation can be supplied by the use of ultrasonic energy, finer droplets with high sphericity and uniform size distribution can be achieved. The other advantage of ultrasound induced atomisation process is the lower momentum associated with ejected droplets compared to the momentum carried by the droplets formed using conventional nozzles. This has advantage in coating and granulation processes. An ultrasonic probe sonicator was designed with a facility for liquid feed arrangement and was used to atomise the liquid into droplets. An ingenious method of droplet measurement was attempted by capturing the droplets on a filter paper (size variation with regard to wicking was uniform in all cases) and these are subjected to image analysis to obtain the droplet sizes. This procedure was evaluated by high-speed photography of droplets ejected at one particular experimental condition and these were image analysed. The correlations proposed in the literature to predict droplet sizes using ultrasound do not take into account all the relevant parameters. In this work, a truly universal correlation is proposed which accounts for the effects of physico-chemical properties of the liquid (flow rate, viscosity, density and surface tension), and ultrasonic properties like amplitude, frequency and the area of vibrating surface. The significant contribution of this work is to define dimensionless numbers incorporating ultrasonic parameters, taking cue from the conventional numbers that define the significance of different forces involved in droplet formation. The universal correlations proposed are robust and can be used for designing ultrasonic atomisers for different applications. Among the correlations proposed here, those ones that are based on the dimensionless numbers and Davies approach predict droplet sizes within acceptable limits of deviation. Also, an empirical correlation from experimental data has been proposed in this work. PMID- 11432435 TI - High power resonant tracking amplifier using admittance locking. AB - A high power resonance tracking ultrasonic amplifier is described. The amplifier is a class D type inverter, configured as a half-bridge in which the output MOSFETs are driven into saturation when on. The resonance tracking system makes use of a new method of frequency locking; admittance locking is used to track the optimum power conversion frequency for the transducer. This new arrangement offers some advantages over phase locking and motional feedback methods. The system is capable of delivering up to 3 kW at up to 25 kHz in resonance tracking operation. PMID- 11432436 TI - Propagation speed of sound assessment in the layers of the guinea-pig esophagus in vitro by means of acoustic microscopy. AB - The study's purpose was to evaluate the propagation speed of sound in the tissue layers of the esophagus at various mechanical loadings. Scanning laser acoustics microscopy was applied for the estimation of the propagation speed in the mucosa submucosa and muscle layers of guinea-pig esophagus in vitro (n = 26). The propagation speed in the esophagus was determined in the no-load state with all external forces removed, and in the distended and zero-stress states. The zero stress state was obtained by cutting the esophageal rings radially. The propagation speed in the no-load state differed significantly (p < 0.001) between the muscle layer (median 1740, quartiles 1735-1746 m/s) and the mucosa (1607, 1605-1609 m/s). In the distended state the propagation speed in the muscle layer decreased significantly (p < 0.001) to 1673 (1666-1681) m/s while it did not change significantly in mucosa (1602, 1600-1607 m/s). When compared to the no load state, the propagation speed in the zero-stress state in the muscle layers decreased to 1624 (1615-1636) m/s (p < 0.001) and in mucosa to 1584 (1566-1603) m/s (p < 0.001). In conclusion, the esophagus is a composite structure with heterogeneous propagation speed characteristics. Furthermore, the mechanical loading state must be considered in esophageal ultrasound studies. PMID- 11432437 TI - Ultrasonic Lamb wave diffraction tomography. AB - Ultrasonic guided waves, Lamb waves, allow large sections of aircraft structures to be rapidly inspected. Unlike conventional ultrasonic C-scan imaging that requires access to the whole inspected area, tomographic algorithms work with data collected over the perimeter. Because the velocity of Lamb waves depends on thickness the travel times of the fundamental modes can be converted into a thickness map of inspected region. Lamb waves cannot penetrate through holes and other strongly scattering defects and the assumption of straight wave paths, essential for many tomographic algorithms, fails. Diffraction tomography is a way to incorporate scattering effects into tomographic algorithms in order to improve image quality and resolution. This work describes the iterative reconstruction procedure developed for Lamb wave tomography and allowing for ray bending correction for imaging of moderately scattering objects. PMID- 11432438 TI - Importance of the near Lamb mode imaging of multilayered composite plates. AB - In recent years Lamb waves are being used for internal defect detection in multilayered composite plates. Different Lamb modes generate various stress levels in different layers. As a result, all Lamb modes are not equally sensitive to internal defects of various layers. A number of studies have been carried out to identify which Lamb mode is most effective for detecting defects in a specific layer. However, one shortcoming of the Lamb wave inspection technique is that in a symmetrically layered composite plate stress and displacement magnitudes and energy distribution profiles for all Lamb modes are symmetric about the central plane of the plate. As a result, the ability of a Lamb mode to detect defects in a specific layer of the plate is identical to its ability to detect defects in the corresponding layer of mirror symmetry. Hence, from the Lamb wave generated image one cannot distinguish between the defects in these two layers of mirror symmetry. In this paper it is investigated how by fine-tuning the frequency and the striking angle of the incident beam in the neighborhood of a Lamb mode one can separately detect internal defects in layers of mirror symmetry in the upper and lower halves of a plate. PMID- 11432439 TI - Combining phase and energy detection with mathematical morphology in dual time frequency representation leads to improved SSP noise robustness. AB - In this paper a fusion algorithm for the detection of specular echoes blurred by coloured Gaussian noise is proposed. The phase and energy criteria, characteristic of these echoes, are handled separately. First, the detection capabilities of split spectrum (phase) and short time Fourier transform spectrogram (energy) are compared theoretically and experimentally. Second, the new algorithm based on mathematical morphology operators using both time frequency representations is proposed. In the last part our method is shown to be more robust than absolute minimisation split spectrum processing. PMID- 11432440 TI - Measurements of dispersion curves of leaky Lamb waves using a lensless line-focus transducer. AB - This paper presents a new method of measuring dispersion curves of leaky Lamb waves propagating in a thin plate. The measurement system is based on a lens-less line-focus transducer and its defocusing measurement technique. A new waveform processing method known as V(f, z) method is introduced for analyzing the measured waveforms and for extracting the dispersion relation. For two thin-plate samples, namely a 150 microm thick alumina plate and a 320 microm thick stainless steel plate, the dispersion curves of several lower order modes of leaky Lamb waves have been experimentally determined in the frequency range of few to 25 MHz. The experimental dispersion curves have then been compared with the theoretical ones. Excellent agreement is observed. It is then verify that this method indeed provides a convenient and accurate way for experimentally measuring dispersion curves of leaky Lamb waves of thin-plate samples. PMID- 11432441 TI - Potentiation of cytotoxicity of adriamycin on human ovarian carcinoma cell line 3AO by low-level ultrasound. AB - The aim of this study was to determine whether the ultrasound, with a dosage that did not lead to acute and delayed inhibition, could potentiate the cytotoxicity of adriamycin to human ovarian carcinoma cell line 3AO in vitro. Drug sensitivity was analyzed by clonogenic assay, cells were treated by adriamycin singly in group ADM (control), adriamycin prior to ultrasound exposure in group ADM + US, and ultrasound irradiation prior to adriamycin administration in group US + ADM. The intracellular drug accumulation in each group was determined by fluorometry. The results were: (1) the values of IC50 were 0.0083, <0.001 and 0.0065 microg/ml in group ADM, ADM + US and US + ADM respectively; the clone surviving rate in group ADM + US and in group US + ADM were decreased (P < 0.001, P < 0.01), compared with control; the surviving rate in group ADM + US was lower than that in group US + ADM (P < 0.01). (2) The intracellular drug accumulations in group ADM + US were promoted (P < 0.01) and not increased significantly in group US + ADM (P > 0.05). These suggested that the low-level ultrasound could enhance the cytotoxicity of adriamycin to human ovarian carcinoma cells and promoted intracellular drug contents played the leading role. PMID- 11432442 TI - Pulsed Doppler flow-line spectrum for focused transducers with apodized apertures. AB - The properties of the single flow-line Doppler spectrum using pulsed wave (PW) ultrasound is studied on the basis of previously developed spectral theory for transducers with apodized apertures. It has been shown previously that the spectral width of Doppler signals from a sample volume in low velocity-shear flow is independent of the sample volume depth but that is not true for the spectra from the individual streamlines. The work presented here on the Doppler flow-line spectrum shows that its width should be invariant with flow-line location, if the sample volume depth is fixed. At the same time, for a transducer operating in PW mode not only the Doppler spectral width depends on the sample volume depth, but also the modal Doppler frequency shift changes with flow-line displacement in the illuminating field except if the sample volume centre and the beam focus coincide. The variation of modal Doppler frequency shift is the more explicit manifestation of the effect of wavefront curvature increasing for lines and sounding depths distant from the focal point. The values of the Doppler shift and spectral bandwidth are reported taking account of beam diffraction and variations in its geometry due to focusing. PMID- 11432443 TI - Resetting baroreceptors to a lower arterial pressure level by enalapril avoids baroreflex mediated activation of sympathetic nervous system by nifedipine. AB - Baroreceptor-unloading-mediated activation of sympathetic nervous system (SNS) by antihypertensive agents, such as dihydropyridine calcium channel blockers (CCB), has been considered to compromise the beneficial effects of the therapy and lead to unsatisfying clinical outcome. The present study was aimed at finding a novel way of using CCB without activating SNS. In anaesthetized Wistar rats, baroreceptor-unloading-mediated reflex activation of SNS, as indicated by tachycardia and increase of plasma catecholamines, was observed after mean arterial pressure (MAP) was decreased by 15 mmHg during 4-h administration of nifedipine, a CCB. However an angiotensin-converting enzyme inhibitor (ACEI), enalapril did not cause tachycardia or increase plasma catecholamine levels when it decreased MAP by 15 mmHg. After 100 min (supposedly baroreceptor resetting or adaptation to hypotension had occurred), enalapril infusion was gradually replaced by nifedipine infusion in 40 min. Nifedipine was infused for another 100 min, which kept the lowered MAP unchanged and did not activate SNS. In anaesthetized spontaneously hypertensive rats (SHR), baroreceptor-mediated reflex activation of SNS was observed after MAP was decreased by 25 mmHg during 4-h nifedipine administration. However enalapril did not cause tachycardia or increase plasma catecholamine levels when it decreased MAP by 25 mmHg. After 100 min, enalapril infusion was gradually replaced by nifedipine infusion in 40 min. Nifedipine was then infused for another 100 min, which kept the lowered MAP unchanged and did not activate SNS. The present study indicated that reflex activation of SNS caused by antihypertensive effect of CCB could be avoided if, prior to CCB administration, baroreceptors have been reset to a lower MAP by a drug that does not activate baroreceptor reflex. PMID- 11432444 TI - Expression of thyrotropin receptors in rat thymus. AB - Thymic hyperplasia is associated with Graves' disease. It has been demonstrated that thyrotropin receptors are expressed in human thymus, and thymic thyrotropin receptors are suggested to be involved in the pathophysiology of Graves' disease. We have studied whether thyrotropin receptors are expressed in rat thymic tissue. Thyrotropin receptor mRNA was demonstrated in 5-day-old, 10-day-old, 20-day-old and adult rat thymus by reverse transcription polymerase chain reaction. Thyrotropin receptor mRNA was also demonstrated in cultured rat thymic epithelial cells. Thyrotropin stimulated cyclic AMP production in cultured rat thymic epithelial cells, suggesting the expression of functional thyrotropin receptors. The present results indicate that thyrotropin receptors are expressed in rat thymus. PMID- 11432445 TI - Mixed exhaled nitric oxide and plasma nitrites and nitrates in newborn infants. AB - Plasma nitrite (NO2-) and nitrate (NO3-) are the stable end-products of endogenous nitric oxide (NO) metabolism. NO is present in the exhaled air of humans, but it is not clear if exhaled NO may be an indicator of the systemic endogenous NO production. The aims of the study were to determine the levels of exhaled NO and plasma NO2-/NO3- in healthy term and preterm newborns, and to assess if exhaled NO correlates with plasma NO2-/NO3- at birth. After the stabilization of the newborn, we measured by chemiluminescence the concentration of NO in the mixed expired breath of 133 healthy newborns. Measurement of exhaled NO was repeated after 24 and 48 hours. Plasma NO2-/NO3- levels at birth were measured by the Griess reaction. NO concentrations were 8.9 (CI 8.1-9.8) parts per billion (ppb), 7.7 (CI 7.2-8.3) ppb and 9.0 (CI 8.4-9.6) ppb at birth, 24 and 48 hours, respectively. At birth, exhaled NO was inversely correlated with gestational age (p=0.008) and birth weight (p<0.001). Plasma NO2-/NO3- level was 27.30 (CI 24.26-30.34) micromol/L. There was no correlation between exhaled NO and plasma NO2-/NO3- levels at birth (p=0.88). We speculate that the inverse correlation between exhaled NO and gestational age and birth weight may reflect a role of NO in the postnatal adaptation of pulmonary circulation. At birth, exhaled NO does not correlate with plasma NO2-/NO3- and does not seem to be an index of the systemic endogenous NO production. PMID- 11432446 TI - Homocysteine thiolactone induces apoptotic DNA damage mediated by increased intracellular hydrogen peroxide and caspase 3 activation in HL-60 cells. AB - The cytotoxicity of homocysteine derivatives on chromosomal damage in somatic cells is not well established. The present study used reactive homocysteine derivative of homocysteine thiolactone (Hcy) to investigate its causal effect on apoptotic DNA injury in human promyeloid HL-60 cells. Our results demonstrated that Hcy induced cell death and features of apoptosis including increased phosphotidylserine exposure on the membrane surface, increased apoptotic cells with hypoploid DNA contents, and internucleosomal DNA fragmentation, all of which occurred in a time- and concentration-dependent manner. Hcy treatment also significantly increased intracellular reactive oxygen species H2O2, which coincided with the elimination of caspase 3 proenzyme levels and increased caspase 3 activity at the time of the appearance of apoptotic DNA fragmentation. Preincubation of Hcy-treated HL-60 cells with catalase completely scavenged intracellular H2O2, thus inhibiting caspase 3 activity and protecting cells from apoptotic DNA damage. In contrast, superoxide dismutase failed to inhibit Hcy induced DNA damage. Taken together, these results demonstrate that Hcy exerted its genotoxic effects on HL-60 cells through an apoptotic pathway, which is mediated by the activation of caspase 3 activity induced by an increase in intracellular hydrogen peroxide. PMID- 11432447 TI - Effect of retinoic acid on glucokinase activity and gene expression in neonatal and adult cultured hepatocytes. AB - It has been shown that all-trans retinoic acid induces prematurely hepatic glucokinase mRNA in ten days-old neonatal rat hepatocytes, however, this effect could be related to the capacity of the retinoid to promote a more differentiated state of the hepatocyte. In this report we demonstrate that physiological concentrations of all-trans retinoic acid stimulate glucokinase activity in both mature fully differentiated hepatocytes and at the onset of the induction of the enzyme in 15 to 17 days-old neonatal hepatocytes. The effects produced by the retinoid were similar both in magnitude and in time, to those elicited by insulin, a well-known stimulator of hepatic glucokinase expression. No additive effect was observed when insulin and retinoic acid were tested together. Using the branched DNA assay, a sensitive signal amplification technique, we detected relative increases in glucokinase mRNA levels of about 70% at 3 and 24 h after the treatment with 10(-6) M all-trans retinoic acid, in both neonatal and adult hepatocytes. These data show that retinoic acid exerts a stimulatory effect on hepatic glucokinase independent of the hepatocyte stage of maturity and suggest a physiological role of retinoic acid on glucose metabolism. The action of retinoic acid on hepatic glucokinase might explain previous observations on the relationship between vitamin A status and liver glycogen synthesis. These findings may serve as basis for further investigations on the biological functions of retinoic acid derivatives on hepatic glucose metabolism. PMID- 11432448 TI - Synergistic antiallergic activity of combined histamine H1- and cysteinyl leukotriene1-receptor blockade in human bronchus. AB - Mast cell histamine (HA) and cysteinyl leukotrienes (CysLT) account for most of the early phase bronchospasm in asthma. However, activation of the smooth muscle CysLT1-receptor plays a major role in asthmatic bronchospasms. CysLT-receptor antagonists or CysLT-synthesis inhibitors are efficacious in asthma but do not completely abolish asthmatic bronchospasms. A recent clinical study showed that combined antagonists loratadine (H1) and zafirlukast (CysLT1) were more effective against allergic bronchospasms than either drug alone. We examined the combined efficacy of H1- and CysLT1-receptor antagonists in allergic human bronchus. The H1- and CysLT1-receptor antagonists chlorpheniramine (CTM; 1 microM) and MK-571 (0.03 microM), were tested alone and in combination, against anti-human IgE antibody (Ab)-induced contractions of passively sensitized isolated human bronchus. Ab-induced allergic contractions were reduced 15% and 36% by CTM (1 microM) and MK-571 (0.03 microM), respectively. Combined CTM (1 microM) and MK 571 (0.03 microM) significantly inhibited the Ab response by 87%. Mechanistic investigations in isolated human bronchus and cultured human cord blood mast cells suggest that H1- and CysLT-receptor interactions likely occur at the airway smooth muscle level. CTM and MK-571 synergistically inhibited human allergic bronchospasm in the present in vitro model. The mechanism underlying this synergistic activity requires further investigation. PMID- 11432449 TI - Melatonin inhibits fatty acid transport in inguinal fat pads of hepatoma 7288CTC bearing and normal Buffalo rats via receptor-mediated signal transduction. AB - Melatonin inhibits fatty acid uptake and linoleic acid-dependent growth in hepatoma 7288CTC in vivo in Buffalo rats. In this study we measured the effects of melatonin on arteriovenous differences for fatty acids across inguinal fat pads in fed and fasted rats to determine if fatty acid transport in white adipose tissue was also affected by melatonin. Intravenous infusion of melatonin in fasted tumor-bearing rats in vivo simultaneously and rapidly inhibited both fatty acid release from fat pads and fatty acid uptake by the tumors. Perfusion of fat pads in situ in normal rats with melatonin (0.1 nM) inhibited fatty acid release (fasted rats) and uptake (fed rats). Fatty acid transport was restored by addition of any of the following: a melatonin receptor antagonist (S 20928, 1.0 nM), pertussis toxin (0.5 microg/ml), forskolin (1 microM) or 8-Br-cAMP (10 microM). We conclude that fatty acid transport in inguinal fat pads requires cAMP and that melatonin inhibits this transport via a melatonin receptor-mediated, Gi protein-coupled signal transduction pathway. Melatonin has both anticachectic and lipid homeostatic actions in the white adipose tissue of inguinal fat pads. PMID- 11432450 TI - Inhibitory effect of resveratrol on proteinuria, hypoalbuminemia and hyperlipidemia in nephritic rats. AB - The effect of resveratrol, a polyphenolic compound present in grapes and other plants, on proteinuria, hypoalbuminemia and hyperlipidemia was studied in rats with glomerulonephritis. The nephritis was induced by an intravenous injection of anti-rat kidney glomerular basement membrane rabbit antiserum. Nephritic rats were given oral intubation of resveratrol (5 mg/day/100 g body weight) for 14 days, while control nephritic rats as well as normal ones were similarly given vehicle alone. By resveratrol treatment, enlargement in liver and kidney due to nephritis induction was significantly reduced, together with partial restoration of nephritis-induced reduction in body weight gain. Both proteinuria and hypoalbuminemia, characteristic symptoms to nephrotic syndrome, were significantly remedied, that is, urinary protein excretion was suppressed and serum albumin concentration was increased by resveratrol treatment. Resveratrol also suppressed significantly hyperlipidemia incident to nephritis, the hypotriglyceridemic action being more prominent than the hypocholesterolemic one. From these results, resveratrol is suggested to be a potent anti glomerulonephritic food factor capable of suppressing proteinuria, hypoalbuminemia and hyperlipidemia at the same time. PMID- 11432451 TI - Cardiac effects of the extract and active components of Radix stephaniae tetrandrae. I. Electrically-induced intracellular calcium transient and protein release during the calcium paradox. AB - The present study was designed to compare the cardiac actions of the extract and individual components, tetrandrine (Tet) and fangchinoline (Fan), of Radix stephaniae tetrandrae (RST). We measured the electrically induced [Ca2+]i transient in single rat ventricular myocytes and protein release following perfusion with a Ca2+ free solution (the Ca2+ paradox) from the isolated perfused rat heart, both of which are known to relate to Ca2+ influx. We found that Tet inhibited both electrically induced [Ca2+]i transient and protein release during the Ca2+ paradox, while Fan had no significant effects. The RST extract containing 9% Tet and 6% Fan by weight also affected the [Ca2+]i transient, and was only slightly, though significantly, less effective/potent than Tet alone. On the other hand, RST extract had a significantly greater inhibitory effect on protein release during the Ca2+ paradox than Tet alone. The observations suggest that the RST extract, which contains a mixture of components, may have more potent effects in the heart than its main active component. PMID- 11432452 TI - Cardiac effects of the extract and active components of radix stephaniae tetrandrae. II. Myocardial infarct, arrhythmias, coronary arterial flow and heart rate in the isolated perfused rat heart. AB - The primary purpose of the present study was to compare the cardioprotective effects of the extract from radix stephaniae tetrandrae (RST) and its individual compounds, tetrandrine (Tet) and fanchinoline (Fan). Secondly, we also compared the cardiac effects of the individual compounds and the RST extract with those of verapamil, a classical Ca2+ channel blocker. The Langendorff isolated perfused rat heart preparation was used. Regional ischaemia and reperfusion was employed to induce myocardial infarct and arrhythmia. Infarct, arrhythmia, heart rate and coronary artery flow were determined in hearts treated with vehicle, RST extract, Tet, Fan, or verapamil. It was found that RST extract, of which only 9% was Tet, and Tet alone produced equally potent ameliorating effects on arrhythmia and infarct induced by ischaemia and reperfusion without further inhibiting ischaemia reduced heart rate and coronary artery flow. Fan had no effects on arrhythmia and infarct induced by ischaemia and reperfusion; but it induced S-T segment elevation and further reduced heart rate and coronary artery flow during ischaemia. Verapamil also ameliorated the effects of ischaemia and reperfusion on arrhythmia and infarct. It should be noted that 1 microM verapamil, that produced comparable effects on infarct and arrhythmia to the RST extract and Tet, further inhibited heart rate during ischaemia. The results indicate that the RST extract produces equally potent cardioprotective and anti-arrhythmic effects as Tet alone. Both RST extract and Tet may be better choices for the treatment of arrhythmia and infarct induced by myocardial ischaemia and reperfusion than the classical Ca2+ channel blocker, verapamil as they do not further reduce heart rate during ischaemia. PMID- 11432453 TI - bcl-2 protects HL-60 cells from apoptosis by stabilizing their intracellular calcium pools. AB - bcl-2 has been shown to enhance cell survival by inhibiting apoptosis induced under different circumstances. In this study we investigated the effects of bcl-2 overexpression on the homeostasis of subcellular organelles such as ER and mitochondria. In our study, HL-60/bcl-2 and control HL-60/neo cells were obtained by transfection of bcl-2 cDNA or the neomycin-resistant gene, respectively. Apoptosis was evaluated by both DNA fragmentation and flow cytometry qualitatively and quantitatively, and the intracellular calcium by Fura-2/AM. Thapsigargin (TG), a highly specific inhibitor of the ER-associated Ca2+ pump, and Br-A23187, a calcium ionophore, were used in this study. Our results showed that overexpression of bcl-2 significantly blocked TG- and Br-A23187-induced apoptosis in calcium containing buffer. Measurement of intracellular calcium showed that bcl-2 overexpression could reduce sustained elevation of cytosolic Ca2+ induced by these agents. However, in calcium-free medium, bcl-2 overexpression maintained Ca2+ uptake in ER of both TG- and Br-A23187-treated cells. Moreover, the depletion of Ca2+ by EGTA enhanced TG- and Br-A23187-induced apoptosis, and reduced the anti-apoptotic action of bcl-2, suggesting that cytosolic Ca2+ elevation may be required for optimal ER pool refilling. These findings suggest that bcl-2 facilitates and maintains the replenishment of Ca2+ in intracellular stores and, as a result, influences the intracellular calcium, thus protecting the cells from death. In addition, there were no cytochrome c release from mitochondria into the cytosol in TG- and Br-A23187- induced apoptosis, suggesting that cytochrome c release is not a universal phenomenon in the apoptotic process. PMID- 11432454 TI - A long-lasting facilitation of hippocampal neurotransmission via a phospholipase A2 signaling pathway. AB - Phospholipase A2, which is linked to a protein kinase C pathway, hydrolyzes phosphatidylcholine into cis-unsaturated free fatty acids and lysophosphatidylcholine (lysoPC). The present study investigated the effect of the free fatty acids, such as arachidonic, oleic, linoleic, and linolenic acid, and lysoPC on neurotransmission by monitoring population spikes (PSs) from the granular cell layer of rat hippocampal slices. All the free fatty acids and lysoPC examined here gradually increased PS amplitude to a different extent, the effect being evident 60 min after treatment. No significant synergistic enhancement in the PS amplitude was not induced by arachidonic acid following oleic acid, linoleic acid or lysoPC. The results of the present study, thus, demonstrate that phospholipase A2-linked free fatty acids and lysoPC are employed in the sustained facilitation of hippocampal neurotransmission, suggesting a significant role of a phospholipase A2 signaling pathway in the neuroplasticity. PMID- 11432455 TI - Fatty acid composition of Pinaceae as taxonomic markers. AB - Following our previous review on Pinus spp. seed fatty acid (FA) compositions, we recapitulate here the seed FA compositions of Larix (larch), Picea (spruce), and Pseudotsuga (Douglas fir) spp. Numerous seed FA compositions not described earlier are included. Approximately 40% of all Picea taxa and one-third of Larix taxa have been analyzed so far for their seed FA compositions. Qualitatively, the seed FA compositions in the three genera studied here are the same as in Pinus spp., including in particular the same delta5-olefinic acids. However, they display a considerably lower variability in Larix and Picea spp. than in Pinus spp. An assessment of geographical variations in the seed FA composition of P. abies was made, and intraspecific dissimilarities in this species were found to be of considerably smaller amplitude than interspecific dissimilarities among other Picea species. This observation supports the use of seed FA compositions as chemotaxonomic markers, as they practically do not depend on edaphic or climatic conditions. This also shows that Picea spp. are coherently united as a group by their seed FA compositions. This also holds for Larix spp. Despite a close resemblance between Picea and Larix spp. seed FA compositions, principal component analysis indicates that the minor differences in seed FA compositions between the two genera are sufficient to allow a clear-cut individualization of the two genera. In both cases, the main FA is linoleic acid (slightly less than one-half of total FA), followed by pinolenic (5,9,12-18:3) and oleic acids. A maximum of 34% of total delta5-olefinic acids is reached in L. sibirica seeds, which appears to be the highest value found in Pinaceae seed FA. This apparent limit is discussed in terms of regio- and stereospecific distribution of delta5 olefinic acids in seed triacylglycerols. Regarding the single species of Pseudotsuga analyzed so far (P. menziesii), its seed FA composition is quite distinct from that of the other two genera, and in particular, it contains 1.2% of 14-methylhexadecanoic (anteiso-17:0) acid. In the three genera studied here, as well as in most Pinus spp., the C18 delta5-olefinic acids (5,9-18:2 and 5,9,12 18:3 acids) are present in considerably higher amounts than the C20 delta5 olefinic acids (5,11-20:2 and 5,11,14-20:3 acids). PMID- 11432456 TI - Characterization of simple and reproducible vascular stenosis model in hypercholesterolemic hamsters. AB - The importance of low-density lipoprotein (LDL) in the etiology of atherosclerosis is well recognized. We have established a reproducible stenosis model in hypercholesterolemic hamsters, and the process of arterial stenosis by thrombus or neointima was studied and compared with that in normal hamsters. The level of plasma LDL was 4.6 times higher in hamsters fed a high-cholesterol diet than in hamsters fed normal food. Endothelial injury in right common carotid arteries was induced using a modified catheter. Arterial blood flow was monitored continuously using a Doppler flow probe. Arterial patency after the initiation of injury in high-cholesterol hamsters was significantly changed as compared with that of normal hamsters. Neointima was observed 2 wk after the vascular injury. The neointimal area of high-cholesterol hamsters was significantly larger than that of normal hamsters. To characterize the stenosis in hypercholesterolemic hamsters, we measured platelet aggregation, thrombin time, activated partial thromboplastin time, and proliferating smooth muscle cells (SMC) in vitro and in vivo. The half-maximal inhibitory concentration value for platelet aggregation induced by thrombin or collagen, the DNA synthesis stimulated by platelet-derived growth factor (PDGF)-BB, and 5-bromo-2-deoxy-uridine labeling indices (proliferating index of SMC in vivo) in high-cholesterol hamsters were each significantly higher than the comparable value from normal hamsters. However, specific binding of PDGF-BB in SMC was not different between the two types of hamsters. Furthermore, we investigated the inhibitory effects of probucol or losartan on neointima formation using this model. Probucol, but not losartan, significantly reduced the neointimal area in hypercholesterolemic hamsters. These findings indicated that high levels of plasma LDL strongly contributed to the development of thrombus and neointima formation via both up-regulation of platelet aggregation and the enhancement of SMC proliferation. This stenosis model may be useful for the investigation of hypercholesterolemia-associated cardiovascular diseases. PMID- 11432457 TI - Effects of dietary defatted squid on cholesterol metabolism and hepatic lipogenesis in rats. AB - Male Sprague-Dawley rats were fed a cholesterol-free (Exp. 1) or cholesterol supplemented (Exp. 2) diet containing 20% casein (control group) or 15% defatted squid and 5% casein (defatted squid group), as protein, for 14 d. Serum and hepatic cholesterol concentrations were lower in rats fed defatted squid than in those fed casein in both cholesterol-free (-20%, P < 0.05 and -15%, P < 0.05, respectively) and cholesterol-supplemented (-25%, P < 0.05 and -15%, P < 0.05, respectively) diets. Hepatic triglyceride concentration was lower in the defatted squid than in the control groups in both cholesterol-free (-51%, P< 0.05) and cholesterol-supplemented diets (-38%, P < 0.01). The activities of cytosolic fatty acid synthase and the NADPH-generating enzymes, malic enzyme and glucose-6 phosphate dehydrogenase, in the liver were lower in the defatted squid than in the control groups in both cholesterol-free (-21%, P< 0.01, -33%, P < 0.05, and 33%, P < 0.01, respectively) and cholesterol-supplemented diets (-34%, P < 0.05, 57%, P < 0.05, and -67%, P < 0.05, respectively). The activity of mitochondrial carnitine palmitoyltransferase in the liver was comparable between the control and defatted squid groups. The activity of Mg2+-dependent phosphatidate phosphohydrolase in the liver cytosol was lower in the defatted squid (-9%, P < 0.05) than in the control groups only in the cholesterol-free diet. Fecal excretion of total steroids was stimulated by the feeding of defatted squid in both cholesterol-free (+77%, P < 0.005) and cholesterol-supplemented diets (+29%, P < 0.01). These results suggest that the nonlipid fraction of squid exerts a hypocholesterolemic effect by increasing the excretion of total steroids in feces. The fraction also induces a triglyceride-lowering activity in the liver by decreasing hepatic lipogenesis. PMID- 11432458 TI - Alpha-tocopherol affects the urinary and biliary excretion of 2,7,8-trimethyl-2 (2'-carboxyethyl)-6-hydroxychroman, gamma-tocopherol metabolite, in rats. AB - In this study, we investigated a change in the excretory content of 2,7,8 trimethyl-2(2'-carboxyethyl)-6-hydroxychroman (gamma-CEHC), a gamma-tocopherol (gamma-Toc) metabolite, in rat urine and bile by using a new high-performance liquid chromatography-electrochemical detection (HPLC-ECD) method. In this determination, CEHC [alpha- and gamma-CEHC, where alpha-CEHC = 2,5,7,8 tetramethyl-2(2'-carboxyethyl)-6-hydroxychroman] in the biological specimens were treated with 3 N methanolic HCl to hydrolyze conjugates and to promote esterification. The methylated samples were extracted by n-hexane/water (1:2). The analyses of the methyl esters of alpha-CEHC and gamma-CEHC were performed by an HPLC-ECD using an ODS-3 column at 35 degrees C. The mobile phase was acetonitrile/water (45:55, vol/vol) containing 50 mM sodium perchlorate. After rat urine and bile samples, respectively, were methylated as described above, methylated biliary metabolites were identified by liquid chromatography-mass spectrometry as methyl esters of gamma-CEHC. Furthermore, we examined the differences in the excretion of gamma-CEHC between rat urine and bile after an oral administration of gamma-Toc or alpha- + gamma-Toc by the above HPLC method. In the gamma-Toc group, each vitamin E-deficient rat was given 0.5 mL of a stripped corn oil preparation containing 10 mg of gamma-Toc. In the alpha- + gamma-Toc group, the rat was given 10 mg of alpha-Toc and 10 mg of gamma-Toc. The content of gamma-CEHC in rat urine from the alpha- + gamma-Toc group was increased more in comparison to the gamma-Toc group at 18-36 h after oral administration. Moreover, the content of gamma-CEHC in rat bile in the alpha- + gamma-Toc group was increased more in comparison to the gamma-Toc group at 6-18 h after oral administration. Therefore, we have suggested that gamma-CEHC was shifted mainly to urinary excretion after gamma-CEHC had been excreted into the bile. Furthermore, we assume that alpha-Toc may affect the metabolism of gamma Toc to gamma-CEHC in the body. PMID- 11432459 TI - Absorption of canthaxanthin by the rat is influenced by total lipid in the intestinal lumen. AB - In this study the effect of luminal lipid on the absorption of canthaxanthin (CTX) was investigated using the lymph duct cannulated rat. Treatments were emulsions designed to deliver increasing amounts of olive oil (10, 30, 50, 70, or 90 mg/h) and CTX (12.5 nmol/h). Emulsions were continuously infused into the duodenum for 12 h, and lymph was collected during the final 6 h of infusion for analysis. As the amount of lipid in the emulsion increased, a linear increase in the absorption of CTX was observed. The recovery of CTX in the lymph when infused with 10 mg/h olive oil was 14.2 +/- 1.2% and with 90 mg/h was 26.9 +/- 5.7%. The efficiency of CTX absorption nearly doubled by increasing the amount of lipid infused with CTX. The correlation between lipid load and CTX absorbed was r= 0.85. We conclude that luminal lipid load affects CTX absorption. PMID- 11432460 TI - Cytotoxic effect of conjugated trienoic fatty acids on mouse tumor and human monocytic leukemia cells. AB - The cytotoxicity of fatty acids from seed oils containing conjugated linolenic acids (CLN) was studied. Fatty acids from pomegranate, tung, and catalpa were cytotoxic to human monocytic leukemia cells at concentrations exceeding 5 microM for pomegranate and tung and 10 microM for catalpa, but fatty acids from pot marigold oil had no effect at concentrations ranging up to 163 microM. The main conjugated fatty acids of pomegranate, tung, catalpa, and pot marigold were cis(c)9,trans(t)11,c13-CLN (71.7%), c9,t11,t13-CLN (70.1%), t9,t11,c13-CLN (31.3%), and t8,t10,c12-CLN (33.4%), respectively. Therefore, the cytotoxicities of fatty acids from pomegranate, tung, and catalpa were supposed to be due to 9,11,13-CLN isomers. To elucidate the cytotoxicity of these CLN, we separated each CLN isomer from the fatty acid mixtures by high-performance liquid chromatography and analyzed its cytotoxicity. The cytotoxicities of c9,t11,c13 CLN, c9,t11,t13-CLN, and t9,t11,c13-CLN were much stronger than that of t8,t10,c12-CLN. Therefore, the higher cytotoxicity of fatty acids from pomegranate, tung, and catalpa than those from pot marigold would be derived from the different activities of 9,11,13-CLN and 8,10,12-CLN. Since there was little difference in the cytotoxicities of c9,t11,c13-CLN,c9,t11,t13-CLN, and t9,t11,c13 CLN, it is suggested that the cis/trans configuration of 9,11,13-CLN isomers had little effect on their cytotoxic effects. The mechanism of the cytotoxicity of the four fatty acids above may involve lipid peroxidation, because the order of toxicity of the fatty acids was consistent with their susceptibility to peroxidation in aqueous phase. This was supported by the decrease in the cytotoxicity of the fatty acids by addition of butylated hydroxytoluene. PMID- 11432461 TI - Effect of sesamin on mitochondrial and peroxisomal beta-oxidation of arachidonic and eicosapentaenoic acids in rat liver. AB - The effects of dietary sesamin on the hepatic metabolism of arachidonic (AA) and eicosapentaenoic (EPA) acids, were investigated with respect to their beta oxidation and secretion as triacylglycerol (TG). For 2 wk, rats were fed three types of dietary oils: (i) corn oil (control) group; (ii) EPA group: EPA ethyl esters/rapeseed oil = 2:3; (iii) AA group: AA ethyl esters/palm oil/perilla oil = 2:2:1, with or without 0.5% (w/w) of sesamin. Dietary sesamin significantly increased the activities of hepatic mitochondrial and peroxisomal fatty acid oxidation enzymes (mitochondrial carnitine acyltransferase I, acyl-CoA dehydrogenase, and peroxisomal acyl-CoA oxidase). Dietary EPA increased mitochondrial carnitine acyltransferase I and peroxisomal acyl-CoA oxidase. Dietary AA, however, had an effect on peroxisomal acyl-CoA oxidase only. In whole liver and the TG fraction, EPA and AA concentrations were significantly increased by dietary EPA and AA, respectively, and were decreased by dietary sesamin. In hepatic mitochondria and peroxisomes, EPA concentration was increased by dietary EPA, but AA was not changed by dietary AA. The addition of dietary sesamin to the EPA-supplemented diet significantly decreased the EPA concentration compared to concentrations found with consumption of dietary EPA alone. These results suggest that sesamin increased beta-oxidation enzyme activities and reduced hepatic EPA and AA concentrations by degradation. The stimulating effect of sesamin on beta oxidation, however, was more significant in the EPA group than in the AA group. Hepatic AA concentration was altered by the joint effect of sesamin through esterification into TG and the stimulation of beta-oxidation. PMID- 11432462 TI - Correlation of fatty acid unsaturation of the major liver mitochondrial phospholipid classes in mammals to their maximum life span potential. AB - Free radical damage is considered a determinant factor in the rate of aging. Unsaturated fatty acids are the tissue macromolecules that are most sensitive to oxidative damage. Therefore, the presence of low proportions of fatty acid unsaturation is expected in the tissues of long-lived animals. Accordingly, the fatty acid compositions of the major liver mitochondrial phospholipid classes from eight mammals, ranging in maximum life span potential (MLSP) from 3.5 to 46 yr, show that the total number of double bonds is inversely correlated with MLSP in both phosphatidylcholine (PtdCho) and phosphatidylethanolamine (PtdEtn) (r = 0.757, P < 0.03, and r = 0.862, P < 0.006, respectively), but not in cardiolipin (P = 0.323). This is due not to a low content of unsaturated fatty acids in long lived animals, but mainly to a redistribution between kinds of fatty acids on PtdCho and PtdEtn, shifting from arachidonic (r = 0.911, P < 0.002, and r = 0.681, P = 0.05, respectively), docosahexaenoic (r = 0.931 and r = 0.965, P < 0.0001, respectively) and palmitic (r = 0.944 and r = 0.974, P < 0.0001, respectively) acids to linoleic acid (r = 0.942, P < 0.0001, for PtdCho; and r = 0.957, P < 0.0001, for PtdEtn). For cardiolipin, only arachidonic acid showed a significantly inverse correlation with MLSP (r = 0.904, P < 0.002). This pattern strongly suggests the presence of a species-specific desaturation pathway and deacylation-reacylation cycle in determining the mitochondrial membrane composition, maintaining a low degree of fatty acid unsaturation in long-lived animals. PMID- 11432463 TI - Cloning and expression of group IB phospholipase A2 isoforms in the red sea bream, Pagrus major. AB - Two cDNA encoding red sea bream DE-1 and DE-2 phospholipases A2 (PLA2) were cloned from the hepatopancreas of red sea bream, Pagrus (Chrysophrys) major. The cDNA of DE-1 PLA2 encoded a mature protein of 125 amino acid residues with an apparent signal peptide of 20 residues and propeptide of 5 residues, and that of DE-2 PLA2, a mature protein of 126 amino acid residues with an apparent signal peptide of 17 residues and propeptide of 6 residues. Comparison of the predicted amino acid sequences for mature DE-1 and DE-2 PLA2 showed that both proteins contain 14 cysteines including Cys 11 and 77 and a pancreatic loop, which are commonly conserved in group IB PLA2; however, the identity in amino acid sequence between DE-1 and DE-2 PLA2 was low (47%). A previous report concerning the cDNA cloning of red sea bream gill G-3 PLA2 and the present results represent the first cloning and sequencing of three distinct isoforms of group IB PLA2 in a single fish species, red sea bream. Reverse transcription-polymerase chain reaction analysis showed that DE-1 PLA2 mRNA was expressed in the hepatopancreas, pyloric ceca, intestine, spleen, gonad, stomach, and kidney, whereas gill G-3 PLA2 mRNA was expressed only in the gills and gonad. The expression of DE-2 PLA2 mRNA was detected in all of the tissues analyzed. These results indicate that three distinct isoforms of group IB PLA2, DE-1 and DE-2 PLA2 in hepatopanceas and gill G-3 PLA2, are expressed in a tissue-specific manner in red sea bream. PMID- 11432464 TI - Inhibitory effects of triterpenoids and sterols on human immunodeficiency virus-1 reverse transcriptase. AB - Fifty-five triterpenoids consisting of 19 tetracyclic, 32 pentacyclic, and 4 incompletely cyclized triterpenoids, and 2 sterols, mostly isolated from various plant and fungal materials, were examined for their inhibitory effects on a purified human immunodeficiency virus type 1 (HIV-1) reverse transcriptase. Twenty triterpenoids and one sterol showed inhibitory effects with 50% inhibition concentration (IC50) values less than 5.0 microM. Among these cycloartenol ferulate (IC50 = 2.2 microM), 24-methylenecycloartanol ferulate (1.9 microM), lupenone (2.1 microM), betulin diacetate (1.4 microM), and karounidiol 29 benzoate (2.2 microM) inhibited most effectively. Some of the triterpenoids and sterols may be potential new lead compounds to find still more potent HIV-1 reverse transcriptase inhibitors. PMID- 11432465 TI - A novel sphingophosphonolipid head group 1-hydroxy-2-aminoethyl phosphonate in Bdellovibrio stolpii. AB - Members of the bacterial genus Bdellovibrio include strains that are free-living, whereas others are known to invade and parasitize larger Gram-negative bacteria. The bacterium can synthesize several sphingophospholipid compounds including those with phosphoryl bonds as well as phosphonyl bonds. In the present study, the dominant sphingophosphonolipid component was isolated by column chromatography, and the long-chain bases, fatty acids, and polar head groups were identified by thin-layer and gas-liquid chromatographic procedures. The definitive structural identity of the sphingolipid was established by nuclear magnetic resonance and mass spectrometry of hydrolysis products and the intact compound. The compound was identified as N-2'-hydroxypentadecanoyl-2-amino-3,4 dihydroxyheptadecan-1-phosphono-(1-hydroxy-2-aminoethane). PMID- 11432466 TI - New glycosphingolipid containing an unusual sphingoid base from the basidiomycete Polyporus ellisii. AB - A new 9-methyl-sphinga-4,8-dienine-containing glucocerebroside (1), together with two additional known analogs, cerebrosides B and D, was isolated from the chloroform-soluble lipid fraction of the ethanol and chloroform/methanol extract of the fruiting bodies of the basidiomycete Polyporus ellisii Berk. and characterized. The structure and relative stereochemistry of the new compound were identified as (2S,3R,4E,8E-1-(beta-D-glucopyranosyl)-3-hydroxy-2-[(R)-2' hydroxyheptadecanoyl]amino-9-methyl-4,8-octadecadiene by means of spectroscopic (1H,13C, and two-dimensional nuclear magnetic resonance; mass spectrometry) and chemical methods. PMID- 11432467 TI - Method to produce 9(S)-hydroperoxides of linoleic and linolenic acids by maize lipoxygenase. AB - Seed from maize (corn) Zea mays provides a ready source of 9-lipoxygenase that oxidizes linoleic acid and linolenic acid into 9(S)-hydroperoxy-10(E),12(Z) octadecadienoic acid and 9(S)-hydroperoxy-10(E),12(Z),15(Z)-octadecatrienoic acid, respectively. Corn seed has a very active hydroperoxide-decomposing enzyme, allene oxide synthase (AOS), which must be removed prior to oxidizing the fatty acid. A simple pH 4.5 treatment followed by centrifugation removes most of the AOS activity. Subsequent purification by ammonium sulfate fractional precipitation results in negligible improvement in 9-hydroperoxide formation. This facile alternative method of preparing 9-hydroperoxides has advantages over other commonly used plant lipoxygenases. PMID- 11432468 TI - Synthetic routes and lipase-inhibiting activity of long-chain alpha-keto amides. AB - Synthetic routes to primary and N-alkyl alpha-keto amides are presented in this paper. Primary alpha-keto amides may be prepared by using an aldehyde as starting material. Commercially available alpha-keto acids may be coupled in high yield with primary amines by the mixed carbonic anhydride method affording N-alkyl alpha-keto amides. Alternatively, N-alkyl alpha-keto amides may be prepared by coupling long-chain alpha-hydroxy acids with amino components, followed by oxidation with pyridinium dichromate or NaOCl in the presence of 4-acetamido 2,2,6,6-tetramethyl-1-piperidinyloxy free radical. The alpha-keto amide derivatives prepared according to these procedures were tested for their ability to form stable monomolecular films at the air/water interface. The inhibition of porcine pancreatic lipase by the alpha-keto amides, spread as mixed films with 1,2-dicaprin, was studied with the monolayer technique. Among the compounds tested in this study, methyl 2-[(2-ketododecanoyl)amino]hexadecanoate was shown to be the most potent inhibitor, causing a 50% decrease in lipase activity at a 0.09 molar fraction. PMID- 11432469 TI - Inhibition of purified pig and human liver retinyl ester hydrolase by pharmacologic agents. AB - Identification of inhibitors of retinyl ester hydrolase (REH) would help to elucidate its role in vitamin A metabolism in vivo. By using standard incubation conditions, the effects of 215 drugs as potential inhibitors of purified pig and human liver REH when acting on micellar substrate retinyl palmitate were evaluated at 16.7, 167, and 1670 microM. Out of the compounds tested, 103 were inhibitors of the pig liver enzyme. The most potent compounds, in order of decreasing activity, were chloral hydrate, lovastatin, phytomenadione, alimemazine, physostigmine, thioridazine, phenoxybenzamine, probucol, cinnarizine, cyclandelate, amiodarone, flupenthixol, and naftidrofuryl; this order is roughly similar to that of their inhibition of human liver REH. Of the 10 tricyclic ring-containing drugs tested, alimemazine was the most potent enzyme inhibitor. The concentrations necessary for 50% enzyme inhibition ranged from <2.6 up to >540 microM. Moreover, inhibitory kinetic studies showed that at least two pharmaceuticals, chloral hydrate and amiodarone, are potent REH inhibitors at therapeutically achievable serum concentrations. First-pass metabolites were inactive as REH inhibitors compared to that of the parent compounds, in the cases of chloral hydrate, lovastatin, and cyclandelate. PMID- 11432470 TI - Clinical use of heparin-coated cardiopulmonary bypass in coronary artery bypass grafting. AB - Clinical handling, risk and benefit of a heparin-coated cardiopulmonary bypass system combined with reduced systemic heparinization in coronary bypass surgery was investigated in a prospective, randomized clinical study. 243 patients (Pts.) were divided into 3 groups: group A (n = 83) had a standard uncoated extracorporeal circulation (ECC) set, and systemic heparin was administered in an initial dose of 400 IE/kg body weight. During ECC activated clotting time (ACT) was kept > or = 480 sec. Group B (n = 77) had the same ECC set completely coated with low-molecular-weight heparin; i.v. heparin was given in the same dose as in group A, ACT was kept at the same level. Group C (n = 83) had the same coated ECC set as group B, but i.v. heparin was reduced to 150 IE/kg, and was set to be > or = 240 sec during ECC ACT. The same circulatory components were used in all 3 groups including roller pumps, coronary suction and an open cardiotomy reservoir. In the postoperative clinical course, recovery was not significantly different between groups, especially with respect to organ dysfunction; but there was significantly reduced postoperative bleeding where heparin-coated ECC and low dose systemic heparinization were both used. This circulatory technique was also associated with a distinctly lower need for postoperative blood replacement. We conclude that heparin-coated extracorporeal circulation combined with either full dose or reduced systemic heparinization can be used effectively with the same standard equipment and procedures as in uncoated technology. Combination with low dose i.v. heparin leads to significantly decreased blood loss and less need for blood replacement. PMID- 11432471 TI - Re-OPCAB vs. Re-CABG for myocardial revascularization. AB - BACKGROUND: The present study compared redo coronary artery bypass grafting (Re OPCAB) techniques with conventional redo coronary artery bypass grafting (Re CABG) with particular focus on myocardial damage and clinical outcome parameters. METHODS: Redo OPCAB (Re-OPCAB) was performed on 20 consecutive patients (15 males, mean age 63.2 +/- 9.3 years) using either the anterolateral approach for minimally invasive direct coronary artery bypass (n = 4) or the Octopus technique with regular sternotomy (n = 16). The Re-CABG group consisted of 20 consecutive patients (18 males, mean age 67.1 +/- 6.6 years). Groups did not differ in the number of atherosclerotic risk factors, or left ventricular, renal or liver function. RESULTS: Duration of surgery, number of bypass grafts and amount of transfused red blood cells did not differ significantly between both groups. Requirement of epinephrine (mg/h) within the first 24 h was lower in the Re-OPCAB group (Re-OPCAB: 0.14 +/- 0.22 vs. CABG: 0.88 +/- 0.97; p<0.01). In addition, CKMB levels at 24 h after operation were lower in the Re-OPCAB group (Re-OPCAB: 10.0 +/- 10.1 vs. Re-CABG: 38.7 +/- 28.1 U/l, p<0.001). There were no acute myocardial infarctions or deaths in the perioperative period. In the CABG group, there was a longer time period to extubation (hours) (Re-OPCAB: 9.8 +/- 3.9 vs. Re-CABG: 28.7 +/- 25.5; p<0.001), and the length of ICU stay was significantly prolonged (OPCAB: 1.3 +/- 0.5 versus Re-CABG: 4.4 +/- 8.7; p<0.001). The graft patency rate at follow-up was 95% in the Re-OPCAB group. CONCLUSION: Re-OPCAB results in decreased cardiac specific enzyme release, reduced requirement of inotropes and comparable clinical outcome in the early postoperative period. It is an appropriate alternative to conventional Re-CABG in selected patients awaiting reoperation for myocardial revascularization. Larger prospective and randomized trials are required to select the appropriate patient who benefits most from one or the other treatment regime. PMID- 11432472 TI - Cell saver, ultrafiltration and direct transfusion: comparative study of three blood processing techniques. AB - BACKGROUND: Intraoperative blood salvage is an important part of blood conservation efforts in cardiac surgery. The purpose of this study is to examine the effects of three different circuit blood-salvaging techniques: centrifugation, ultrafiltration and direct infusion. METHODS: Sixty patients undergoing elective coronary bypass graft procedures were randomly assigned in a prospective manner to one of the three blood-salvaging methods. RESULTS: Intra- and postoperative blood samples demonstrated increased hemoglobin values in the direct infusion group and higher platelet count in the ultrafiltration group. There were no significant differences in these results. The analysis of coagulation parameters revealed a similar prolongation of partial thromboplastin time and activated clotting time in all groups. The amount of blood loss was not significantly different between the three blood conservation methods. The effect of direct infusion method does not result in either major disturbance of coagulation parameters or in increased blood loss. CONCLUSION: In a sample of adult patients undergoing coronary artery bypass grafting, direct transfusion is, in consideration of the cost-effective factor, an appropriate approach for returning cardiopulmonary bypass circuit blood. PMID- 11432473 TI - The influence of the duration of cardiopulmonary bypass on coagulation, fibrinolysis and platelet function. AB - BACKGROUND: The duration of cardiopulmonary bypass (CPB) might influence blood coagulation. This appears particularly relevant in the light of new, less invasive techniques that propose smaller incisions at the expense of a possible prolongation of time on CPB. METHODS: The time-dependent effects on coagulation, fibrinolysis and platelet function were investigated in 94 patients scheduled for elective coronary artery bypass grafting. Tests on coagulation, fibrinolysis, and platelet function (flow cytometric assay of expression densities of glycoprotein IIb/IIIa and P-selection were performed the day before surgery and after completion of surgery. RESULTS: A significant correlation was found between the duration of CPB and parameters of increased coagulation, decrease of platelet counts during CPB and platelet function. Longer duration of CPB led to an increased need for transfusion of red blood cells. CONCLUSIONS: The duration of CPB affects thrombin formation as well as platelet count and function, but not the fibrinolytic system. This may prove to be a disadvantage when employing minimally invasive techniques that prolong the duration of CPB. PMID- 11432474 TI - Hirudin protects from leukocyte/endothelial cell interaction induced by extracorporeal circulation. AB - BACKGROUND: The clinical complications of Extracorporeal Circulation (ECC) have been linked to disturbances in the microcirculation. In order to prevent these deleterious effects, a biodegradeable agent to coat the extracorporeal circuit was tested. METHODS: Intravital fluorescence microscopy was used on the hamster skinfold chamber model in permanently instrumented, awake animals. ECC was introduced via a micro-roller-pump and a silicon tube shunted between the carotid artery and the jugular vein. The ECC-tube system was coated with PEG-Hirudin Iloprost, two additional groups received either Iloprost i.v. (0.8 mg/kg/h) or Hirudin i.v. (1 mg/kg b.w.). RESULTS: ECC for 20 minutes resulted in an increase in rolling and adherent leukocytes in postcapillary venules (Roller 9 to 36 [%]; Sticker 24 to 330 [n/mm2]). Use of the coated tube system reduced L/E cell interaction (Roller 9 to 24* [%], Sticker 28 to 194* [n/mm2]; *p<0.05), whereas Hirudin i.v. nearly abolished it. CONCLUSIONS: The protective effects of the coating and of Hirudin i.v are probably a result of an attenuated activation of the coagulation-fibrinolytic system. PMID- 11432475 TI - Online detection of myocardial ischemia by near infrared spectroscopy with a fiberoptic catheter. AB - BACKGROUND: The interruption of myocardial O2-supply in acute myocardial ischemia causes differences in coronary venous O2-content. In this study, the oxygenation status of coronary sinus blood is detected in myocardial ischemia by near infrared (NIR) spectroscopy. METHOD: For the intravascular application, a new fiberoptic catheter is developed. After calibration of the catheter by perfusion of blood with standardized gas mixtures in an in vitro experiment, the in vivo application was performed in 12 domestic pigs. The catheter was placed into coronary sinus and an anterior ischemia was induced by temporary occlusion of LAD. NIR graphs and hemodynamic data were obtained for 90 min ischemia and 90 min reperfusion time. RESULTS: Main NIR-spectroscopic differences between oxygenated and de-oxygenated hemoglobin took place in O2-concentrations less than 30%. Coronary sinus NIR spectra of hemoglobin-oxygen binding showed significant, reproducible differences between pre-ischemia, LAD-occlusion and reperfusion. NIR graphs also show variations related to CO2-concentration, pH and temperature. CONCLUSION: The intravascular application of NIR spectroscopy could be a reliable tool in detection and follow-up of acute myocardial ischemia and infarction. PMID- 11432476 TI - The role of endothelin-B receptors in myocardial and endothelial reperfusion injury after heart transplantation. AB - BACKGROUND: This study was designed to investigate the effects of the selective endothelin-B (ET-B) receptor agonist IRL1620 and the selective ET-B receptor antagonist BQ788 on myocardial and endothelial function after reversible deep hypothermic ischemia and reperfusion. METHODS: Isogenic intraabdominal heterotopic heart transplantation was performed on Lewis rats. After one hour of cold ischemic preservation, reperfusion was started either after application of saline vehicle or IRL1620 or BQ788 or both. Left ventricular pressure-volume relations and myocardial blood flow were assessed after one and 24 hours of reperfusion. Endothelium-dependent vasodilatation to acetylcholine (ACH) and endothelium-independent vasodilatation to sodium nitroprusside (SNP) were also determined. RESULTS: IRL1620 attenuated and BQ788 improved myocardial contractility significantly as indicated by the left or upward shift of the systolic pressure-volume relation, respectively, and significantly changed myocardial blood flow during early reperfusion (p<0.05). Although myocardial function and baseline myocardial blood flow were similar in both groups after 24 hours of reperfusion, endothelium-dependent vasodilatation was still significantly lower in the IRL1620 and higher in the BQ788 group (p<0.05). CONCLUSIONS: These results suggest that activation of the ET-B receptors contributes to reperfusion injury after cardiac preservation in a rat heart transplant model. PMID- 11432477 TI - Incidence and impact of systemic venous collateral development after Glenn and Fontan procedures. AB - BACKGROUND: Development of systemic venous collaterals after Glenn or Fontan procedures can lead to systemic desaturation and reduction in ventricular function, resulting in impaired everyday performance in patients with univentricular heart disease. METHODS: We analyzed 79 patients who had undergone a Glenn or Fontan procedure between 1995 and 1999 for the incidence and predilection sites of systemic venous collaterals as well as the therapeutic options. RESULTS: In 16/79 (= 20.2%) patients, 19 veno-venous connections were detected 310 days (1-966 days) postoperatively. Locations were: brachiocephalic angles/pericardial veins (7), azygos/hemiazygos system (5), Thebesian veins (2), epidiaphragmatic veins (5). Drainage was to the pulmonary veins in 5, to the "left" atrium in 9, and to the IVC system in 5 patients. An isolated intervention became necessary because of low saturations in 5/16 pts, with improvement in all of them (catheter embolization 4, surgical closure 1). CONCLUSIONS: After Glenn or Fontan operations, the increased central venous pressure may induce recanalization of embryologically preformed and obliterated vessels. Their predilection sites must be carefully evaluated pre- and postoperatively. During surgical procedures, potential venous channels should be ligated. Interventional or surgical closure of collaterals may become necessary. PMID- 11432478 TI - Serum S-100beta protein release in coronary artery bypass grafting: laminar versus pulsatile flow. AB - BACKGROUND: Cerebral injury after Cardiopulmonary bypass (CPB) is still a serious and unpredictable complication. The S-100beta serum marker has been suggested as potentially useful in the detection of cerebral injury during and after CPB. Direct comparisons of whether laminar or pulsatile pump flow in CABG leads to higher S-100beta values and which type might be more neuroprotective have not been made so far. METHODS: All 21 patients of the study were undergoing CABG for the first time and had no history of cerebral disease in whatever form. They were divided into two groups: laminar (n = 10) versus pulsatile (n = 11) pump flow. In all cases, a Stockert roller pump (Fa. Stockert, Munich, Germany) with a laminar and pulsatile running mode was used for cardiopulmonary bypass. Serum S-100beta levels were detected using a monoclonal immunoradiometric assay (Sangtec Medical AB, Bromma, Sweden). In total, 5 different samples were drawn per patient, starting before intubation and ending 36 hours after surgery. RESULTS: S-100beta peak values were found at skin closure. Median levels were lower in the pulsatile group. Due to the small study group and wide range, results are non-significant. CONCLUSION: The results indicate that pulsatile flow might have a more neuroprotective effect than laminar flow as S-100beta values were lower. PMID- 11432479 TI - Carcinoma arising in congenital lung cysts. AB - We report on a patient presenting with a bronchioloalveolar carcinoma fortuitously detected in the wall of a bronchogenic cyst. Evidence suggests that unstable epithelial cells contained within the cyst wall may lead to premalignant proliferation and neoplasia. In the current case, we demonstrated an increased proliferative activity in some areas of the cyst consistent with atypical adenomatous hyperplasia. Hence, we stress the importance of close follow-up of all suspected congenital lung cysts because of their potential malignant degeneration. PMID- 11432480 TI - Paraplegia as an unusual manifestation of aortic coarctation. AB - In this report, we present an unusual case of a 20-year-old man whose first symptom of aortic coarctation was sudden paraplegia due to spinal epidural hemorrhage caused by rupture of an aneurysmal collateral vessel. Now, one year after surgical correction of coarctation, the patient has had no cardiac or neurological problems. To our knowledge, this is the only clearly documented case of such an aortic coarctation complication. PMID- 11432481 TI - Pneumonectomy for lung cancer in heart transplant recipient. AB - Prolonged nonspecific immunosuppression after solid-organ transplantation is associated with an increased risk of certain cancers. Review of the medical literature reveals that the combination of profound immunossuppression (triple drug immunosuppression), a heavy smoking history, advanced age and a working exposition puts cardiac transplant recipients at increased risk for the development of aggressive lung cancer. These tumors in cardiac transplant recipients carry a poor prognosis. We present one case of bronchogenic carcinoma in a cardiac transplant patient. The patient was operated to resect the tumor and a long-term cure and a good quality of life should, however, be offered. PMID- 11432482 TI - Scope-guided implantation of a stented elephant trunk in acute aortic dissection. AB - A reliable guide is essential for implanting a stented graft safely into a recently dissected, fragile aorta. In 4 patients with acute aortic dissection, the implantation of a stented elephant trunk was done safely using an endoscope for direct visualization. In all patients, the operation went well. The placement of a stent appears to enhance the benefit of the elephant trunk, which itself reduces the complications of an arch replacement in acute dissection. PMID- 11432483 TI - A forgotten gauze swab--clinical and legal considerations. AB - In this report, we will present two cases of retained gauze swabs following open heart surgery. We will discuss diagnostic and treatment strategies as well as medicolegal implications with regard to individual liability and possible penalties according to the German law. PMID- 11432484 TI - Fluoride content of infant formulas and market milk in Turkey. AB - The aim of the present study was to test the fluoride contents of infant formulas and market milk in Turkey. Fifteen formulas and nine market milks were prepared according to manufacturers' instructions with deionized water. Fluoride contents were analyzed with a spectrophotometer (CADAS 50S) and results were obtained as ppm (microgF/ml). Mean fluoride content of the formulas was 0.101 ppm F and of the milks was 0.08 ppm F. Formulas reconstituted with water containing < 0.3 ppm F do not provide a daily fluoride intake at any age. To decrease dental caries among children in Turkey, fluoride supplements could be prescribed at minimal dosages according to the ADA fluoride schedule. PMID- 11432485 TI - Monotherapy with meropenem versus combination therapy with piperacillin plus amikacin as empiric therapy for neutropenic fever in children with lymphoma and solid tumors. AB - The purpose of this study was to compare meropenem monotherapy with combination therapy for empirical treatment of neutropenic fever in children with lymphoma and solid tumors. Ninety episodes of neutropenic fever in children (0.7-16.0; mean age 7.7 years) with solid tumors in a single center were randomized to receive either meropenem (50 mg/kg/dose-maximum 1 g, every 8 hours) or piperacillin (200 mg/kg/dose, every 6 hours) plus amikacin (15 mg/kg daily). Failure was defined as treatment modification. Non-Hodgkin's lymphoma (NHL) accounted for 62.2 percent of all episodes, and solid tumors (37.8%) for the rest. Blood cultures were positive in 23 percent of all episodes. Sixty-seven percent of all isolated microorganisms stained Gram-positive. Overall success was 70.0 percent (63/90). The success with meropenem was comparable to that seen with piperacillin plus amikacin: 76.6 versus 64.6 percent (p = 0.25). The failure rate was 33 percent with Gram-positive culture and 78 percent with Gram-negative or mixed cultures. The solid tumor group had significantly less bacteremia (4/34 versus 17/56; p < 0.05) and treatment failure (3/34 versus 24/56; p < 0.001) than the NHL group. No serious drug-related adverse event was noticed. Meropenem monotherapy was as effective as piperacillin plus amikacin combination in the empirical treatment of neutropenic fever in children with lymphoma and solid tumors. PMID- 11432486 TI - Molecular genetic analyses of cystinuria type 1 in 24 Turkish patients. AB - M467T mutation (exon 8) in rBAT gene is found to be the most common mutation in cystinuria type I patients. In our series consisting of 24 patients, the allele frequency of the M467T mutation was 8.3 percent (4/48). The second most frequent mutation at the same nucleotide position was M467K, with an allele frequency of 4.2 percent (2/48). The polymorphism which is found in linkage disequilibrium with the M467T is 231T/A (exon 1). We also found that 231T/A was associated with the M467T mutation in our series. PMID- 11432487 TI - A practical GnRH analogue (triptorelin) stimulation test to distinguish constitutional delay of puberty from hypogonadotropic hypogonadism in prepubertal boys. AB - At present, no established practical and reliable endocrine test exists for differentiating between constitutional delay of puberty (CDP) and hypogonadotropic hypogonadism (HH). The most discriminating results have been reported by measuring LH and testosterone (T) responses to gonadotropin-releasing hormone (GnRH) analogues. In this study, 23 prepubertal boys aged 14 to 16.5 years underwent a modified triptorelin (a GnRH analogue) stimulation test, and they were followed clinically for up 24 to months. Sixteen subjects developed spontaneous puberty during the follow-up period and thus were diagnosed with CDP, and the remaining seven were diagnosed with HH. Retrospective evaluation of their LH, FSH and T responses revealed significant differences without any overlaps in serum LH levels at 4 h (CDP: 33.2 +/- 9.3 vs. HH: 3.3 +/- 2.6 mIU/ml, p < 0.0002) and in serum T levels at 24 h (CDP: 369.3 +/- 128.1 vs. HH: 61.4 +/- 22.6 ng/dl p < 0.0002). We conclude that CDP can be clearly differentiated from HH by the LH response at 4 h and/or T response at 24 h after a single-dose triptorelin administration. PMID- 11432488 TI - Breast milk beta-glucuronidase levels in hyperbilirubinemia. AB - Breast milk beta-glucuronidase was thought to be one of the etiological factors in the pathogenesis of late-onset breast-milk jaundice, but results of these studies are conflicting. In this study breast milk beta-glucuronidase levels were determined in groups with physiologic jaundice, early breast-feeding jaundice and late breast-milk jaundice. No difference in beta-glucuronidase levels of these three groups was found in samples taken on the 4th and 15th days of life. beta glucuronidase activity in breast milk declined from the 4th to 15th day in all groups. These results imply that factors other than breast milk beta glucuronidase activity should be investigated to reveal the pathogenesis of late onset breast-milk jaundice. PMID- 11432489 TI - Serum ferritin, iron levels and iron binding capacity in asymmetric SGA babies. AB - The concentration of serum ferritin reflects the extent of iron stores in premature infants. We aimed to determine serum ferritin levels and iron status in asymmetric small for gestational age (SGA) babies. This study was performed on 21 SGA babies and 19 appropriate for gestational age (AGA) babies. Hemoglobin, iron, iron binding capacity and ferritin levels were investigated in the first six hours after the birth. Hemoglobin levels in the SGA and control groups were 20.9 +/- 1.3 (19.4-23.4 g/dl) and 19.6 +/- 0.8 (18.5-21.5 g/dl), respectively (p = 0.001). Serum ferritin levels in the SGA and AGA groups were 58.36 +/- 20.1 ng/ml and 90.46 +/- 30.5 ng/ml, respectively. Ferritin levels were found lower in the SGA group (p < 0.001). In the SGA group, decreased serum iron and increased iron binding capacity were found but the difference was not significant (p > 0.05). Decreased ferritin levels may result from either impaired iron transport associated with uteroplacental vascular insufficiency or increased iron utilization during enhanced erythropoiesis in conditions characterized by chronic fetal hypoxia. Our results stress the significance of iron supplementation and careful anemia follow-up in term SGA babies. Because anemia progress early, beginning iron therapy as soon as possible is a necessity in SGA babies as in prematures. PMID- 11432490 TI - Seroprevalence of Helicobacter pylori in a pediatric population. AB - Helicobacter pylori (H. pylori) is one of the common bacterial infections in humans. In this study, seroprevalence of H. pylori infection in a pediatric population in Izmir and its relationship with different variables were investigated. Two hundred twenty-six children (115 boys, 111 girls, age range: 1 18) were tested for anti-H. pylori IgG. Socioeconomic conditions, living area (urban or rural), and number of people living in the same house were noted for each subject. H. pylori antibodies were determined by an enzyme immunoassay. Overall, 120 (53%) subjects were seropositive for H. pylori. The seroprevalence of H. pylori increased significantly with age and poor socioeconomic conditions. Seroprevalence did not differ according to sex, number of people living in the same house or living area. PMID- 11432491 TI - Critical learning period for speech acquisition and screening techniques in early detection of hearing impairment. AB - In order to develop verbal communication skills, an infant nervous system needs sound stimuli, especially human speech, in the early and critical learning period of life. The maturation of hearing and language learning abilities of an infant is considered to be complete within the first two to three years of life. Therefore, detection of hearing impairment before the end of the critical language learning period is considered crucial if a child is expected to acquire his/her native language properly. Parents or caregivers may suspect the presence of a delay in language acquisition; however, relying only on parental awareness could cause a delay in detection of the hearing impairment. Even mild hearing loss may interfere with normal development of speech and language in infants, therefore, universal screening of infants for hearing loss as early as possible should be the goal. The purpose of this paper is to discuss the concept of the critical language learning period and its relevance to language acquisition. Furthermore, it is the aim of the paper to evaluate the available techniques for early identification of hearing impairment in general and the feasibility of the various techniques in Turkey. PMID- 11432492 TI - Apoptosis in renal disease: a brief review of the literature and report of preliminary findings in childhood lupus nephritis. AB - Apoptosis, a programmed form of cell death, is an important mechanism that maintains cellular homeostasis. The cellular content of tissues is regulated by a balance between cell proliferation and cell loss. Apoptosis is important not only in physiological conditions but in pathological processes as well. Apoptosis has been implicated in the pathogenesis of certain renal diseases. In human models, systemic lupus erythematosus (SLE) and IgA nephropathy have been the main interests. These studies have mainly shown that apoptosis is important in the control of mesangial cell population. We have attempted to define the role of apoptosis in a cohort of childhood lupus nephritis. We have analyzed apoptosis by the terminal deoxynucleotidyl transferase (Tdt)-mediated dUTP-biotin nick end labeling (TUNEL) method in eight SLE pediatric patients, two of whom had hereditary deficiencies of complement components. Although the sample size was small because of the rarity of hereditary complement deficiencies, we have shown that apoptotic activity was the greatest among these pediatric patients. It has been previously suggested that in lupus, autoimmunity develops as a result of inadequate clearance of apoptotic blebs containing nuclear elements; complement deficiencies are the most important hereditary factors predisposing to the inadequate clearance of apoptotic particles. This is the first time this hypothesis has been evaluated in the tissue samples of hereditary complement deficiency-related proliferative lupus nephritis. On the other hand, apoptosis was not different from the other mesangial proliferative glomerulopathies in the lupus nephritis samples. Further studies are needed to confirm our preliminary findings. Apoptosis has been implicated in other renal diseases as well, such as autosomal polycystic kidney disease, and in experimental models. A short review of the relevant literature is presented highlighting the role of apoptosis in the pathogenesis and prognosis of certain renal diseases. PMID- 11432493 TI - Lupus anticoagulant and protein S deficiency in a child who developed disseminated intravascular coagulation in association with varicella. AB - Varicella is not always a benign disease it may cause serious complications. We report a two-year-old boy with disseminated intravascular coagulation in association with varicella. The patient had the lupus anticoagulant, the antiphospholipid antibody, acquired free protein S deficiency, and increased concentrations of the prothrombin F 1+2 fragment. Intravenous immunoglobulin was administered due to its potential antibody-blocking activity, and the patient responded well. We recommend that children with varicella and disseminated intravascular coagulation should be examined for the lupus anticoagulant, the free protein S antigen, the prothrombin fragment F 1+2 and the other coagulation parameters. Intravenous immunoglobulin administration could be useful in such conditions because of its antibody-blocking activity. PMID- 11432494 TI - Dermatomyositis with membranous nephropathy. AB - Dermatomyositis is the connective tissue disease with the least renal involvement. Although some renal findings like proteinuria, hematuria, pyuria, progressive renal insufficiency, and glomerular and tubular calcium deposits with arteriolar fibrosis have been described, glomerulonephritides have rarely been associated with dermatomyositis, especially in childhood cases. We describe a 10 year old boy with the clinical picture of dermatomyositis who underwent renal biopsy due to microscopic hematuria demonstrating membranous glomerulonephritis with Clq deposition. Children with "full-house" membranous glomerulonephritis with deposition of Clq and the other immunoglobulins have been reported to present in the future with the clinical findings of systemic lupus erythematosus. However, laboratory evaluation of our patient for systemic lupus erythematosus was negative at the present time. Thus, we think this case should be followed up closely with special attention to the possible clinical and laboratory findings of systemic lupus erythematosus. PMID- 11432495 TI - Central nervous system involvement of polyarteritis nodosa: a case report. AB - Polyarteritis nodosa (PAN) is a necrotizing vasculitis involving small and medium sized arteries and it affects multiple organ systems in the body Central nervous system (CNS) involvement appears less frequently, and usually develops after the disease is established. Although aneurysms are common in visceral arteries in PAN, intracranial aneurysms are uncommon and have been documented rarely. This case is reported to raise awareness among radiologists as it has characteristic and rare, if not specific, imaging findings of CNS involvement of PAN. PMID- 11432496 TI - Rapidly progressive bronchiectasis complicating ulcerative colitis in a child. AB - Patients with ulcerative colitis may have a presentation dominated by extraintestinal manifestations. These manifestations, particularly bronchiectasis, are very rarely seen in pediatric patients. A 13-year-old boy with ulcerative colitis who was diagnosed by colonic mucosa biopsy is presented. He developed unexplained productive cough after the appearance of colonic disease. He was treated and followed up at his primary care hospital with the sole diagnosis of ulcerative colitis, with little attention given to the chest symptoms. The relation of the bronchial involvement to the ulcerative colitis was not established until two years after the onset of disease. Thoracal computed tomography (CT) examination after this period showed evidence of bronchiectasis and pulmonary involvement. Despite prophylactic inhaled corticosteroid treatment, no clinical or radiographic improvement was observed and widespread bronchial destruction developed very rapidly. More effective treatment with oral steroids was probably necessary in this patient, if the early chest symptoms were related to the ulcerative colitis. PMID- 11432497 TI - Unilateral late-onset tibia vara associated with bilateral proximal femoral growth disturbance in monozygotic twins: case report. AB - Clinical and histopathological similarities and rare association of Blount's disease with various proximal femoral physeal affections (i.e. adolescent coxa vara and slipped capital femoral epiphysis) are well known. Association of tibia vara with another epiphyseal disease of the proximal femur has not been reported previously. In this paper, a monozygotic set of twins with concordant bilateral epiphyseal growth disturbance of the proximal femur and unilateral late-onset tibia vara is presented. Radiological characteristics of the affected knees revealed a wedging in the proximal tibial epiphysis, depression of the medial joint surface and varus deformity of the tibia. Proximal femurs of both cases showed aspheric congruity, coxa magna, shortness of the femoral neck, and subchondral cystic changes. The presented cases support the genetic etiology of tibia vara, and association of the two conditions is unique. PMID- 11432498 TI - Cutis marmorata telangiectatica congenita: an unusual cause of lower extremity hypoplasia. AB - Cutis marmorata telangiectatica congenita is a relatively benign, rare cutaneous disorder with various manifestations. A seven-year-old girl, who presented with extremity hypoplasia, had the characteristic reticular, patchy, blue-pink cutaneous lesions, which became more prominent with exposure to cold temperatures. She had 4.8 cm shortening of her right lower extremity, which was also thinner than on the left side. The patient did not have skin atrophy, ulcers, glaucoma or macrocephaly. She is being followed for a future extremity lengthening procedure. PMID- 11432499 TI - Chronic idiopathic neutropenia associated with gingival enlargement. AB - A girl with chronic idiopathic neutropenia who developed gingival enlargement at seven years of age is presented. Intraoral examination revealed generalized gingival inflammation with a tendency to bleeding and inflammatory gingival enlargement localized to the anterior region. A considerable amount of bacterial plaque was noted on the teeth. There were also 4-5 mm pocket depths around the first molars. Radiographic examination also indicated the presence of incipient bone loss around the first molars in both jaws. The patient, who was diagnosed as localized prepubertal periodontitis with generalized gingival inflammation and anterior gingival enlargement, accentuates the importance of evaluation of periodontal status in patients with chronic idiopathic neutropenia, to avoid the destruction of supporting structures of the dentition. PMID- 11432500 TI - A case with some clinical findings overlapping to Rubinstein-Taybi, Rubinstein Taybi-like syndrome or multiple pterygium syndrome: coincidental findings or a new entity? AB - We report a case with broad, deviated thumb and big, duplicated, deviated toes resembling Rubinstein-Taybi syndrome. But the patient did not have severe mental retardation as in Rubinstein-Taybi syndrome and had no microdeletions on chromosome 16 by FISH-based assay. This patient had mild webbing as seen in multiple pterygium syndrome, but broad-deviated thumb has not been reported in this syndrome. We discuss whether these are coincidental or overlapping findings or whether this is a possible new clinical entity. PMID- 11432501 TI - Bleomycin-induced hyperpigmentation and hypersensitivity reactions to etoposide and vinblastine in a child with endodermal sinus tumor. AB - We report a pediatric case who developed bleomycin-induced hyperpigmentation and hypersensitivity reactions to both etoposide and vinblastine while receiving chemotherapy for germ cell tumor. Skin hyperpigmentation related to chemotherapeutic agents has been reported only rarely in pediatric patients. This patient developed a characteristic skin hyperpigmentation which was "flagellate" in appearance. Two features of the hyperpigmentation were noteworthy: development at a low cumulative dose of bleomycin and persistence after cessation of chemotherapy. Additive effect of cisplatinum-induced hyperpigmentation was suggested. Although hypersensitivity reactions to etoposide have been previously reported, hypersensitivity reactions to vinblastine are almost unknown. To our knowledge, this is the first report of hypersensitivity reaction to vinblastine in a child in English literature. PMID- 11432502 TI - Subadventitial hematoma of a patent ductus arteriosus and thrombus formation within the aorta secondary to cardiac catheterization. AB - An 18-month-old boy with patent ductus arteriosus (PDA) underwent surgical closure of PDA. Cardiac catheterization determined the PDA was not suitable for transcatheter closure. After the chest was opened, subadventitial hematoma was seen on the aortic end of the PDA. Incision of the aorta revealed a thrombus secondary to intimal laceration. The thrombus was extracted and the PDA was closed using division technique. Because no similar report was found in English language literature, the technique and the surgical strategy are discussed. PMID- 11432503 TI - Global fibrinolytic capacity in healthy newborn infants. PMID- 11432504 TI - Neutrophil elastase gene mutations in cyclic neutropenia. PMID- 11432505 TI - Newborn PKU screening in Turkey: at present and organization for future. AB - At present, pkenylketonuria screening is a national child health program in Turkey which is carried out collaboratively by the Ministry of Health and three University Children's Hospitals in Ankara, Istanbul and Izmir. Since 1986 the number of cities included in the screening program has gradually increased, now and it covers all the metropolises the country. A total of 383 babies were found with persistent hyperphenylalaninemia (1:4,172) among 1,605,582 babies screened by the Guthrie test at the Hacettepe Screening Center in Ankara. By taking into account pretreatment phenylalanine levels and phenlyalanine tolerances at five years of age, the numbers of classical and mild-moderate phenylketonuria and mild hyperphenylalaninemia cases were 216, 102 and 58, respectively. The major problems encountered in the screening program and in management of the detected cases were unsatisfactory sample collection, early discharge from maternity hospitals, difficulties in reaching some detected cases, and noncompliance with dietary therapy due to illiterate parents or to lack of social insurance. To screen and treat all newborns for phenylketonuria and to include at least hypothyroidism in the screening program, there is a need for a more disciplinary intersectoral approach than exists at present. PMID- 11432506 TI - Serrated adenomas. PMID- 11432507 TI - Barrett's oesophagus--time for a reality check? PMID- 11432508 TI - Prevalence of intestinal metaplasia in the distal oesophagus, oesophagogastric junction and gastric cardia in symptomatic patients in north-east Italy: a prospective, descriptive survey. The Italian Ulcer Study Group "GISU". AB - BACKGROUND: Incidence of adenocarcinoma of distal oesophagus and gastric cardia, probably arising from areas of intestinal metaplasia, has been increasing rapidly. AIMS: To define prevalence of intestinal metaplasia of distal oesophagus, oesophagogastric junction and gastric cardia and to evaluate potential associated factors, by means of a prospective multicentre study including University and teaching hospitals, and primary and tertiary care centres. PATIENTS: Each of 24 institutions involved in study enrolled 10 consecutive patients undergoing first-time routine endoscopy for dyspeptic symptoms. METHODS: Patients answered symptom questionnaires and underwent gastroscopy Three biopsies were taken from distal oesophagus, oesophago-gastric junction and gastric cardia, and were stained with haematoxylin and eosin. Specimens were also evaluated for Helicobacter pylori infection. RESULTS: A total of 240 patients (124 male, 116 female; median age 56 years, range 20-90) were enrolled in study. Intestinal metaplasia affected distal oesophagus in 5, oesophago-gastric junction in 19 and gastric cardia in 10 patients. Low-grade dysplasia was found at distal oesophagus and/or oesophago-gastric junction of 3/24 patients with intestinal metaplasia vs 2/216 without intestinal metaplasia (p<0.05). A significant association was found between symptoms and presence of intestinal metaplasia, regardless of location, and between Helicobacter pylori infection and intestinal metaplasia at oesophago-gastric junction. CONCLUSIONS: Intestinal metaplasia of distal oesophagus, oesophagogastric-junction and gastric cardia is found in a significant proportion of symptomatic patients undergoing gastroscopy and is associated with dysplasia in many cases. Although prevalence of dysplasia seems to decrease when specialized columnar epithelium is found in short segment, or even focally in oesophago-gastric junction, these small foci of intestinal metaplastic cells may represent source of most adenocarcinomas of cardia. PMID- 11432509 TI - Prevalence of short-segment Barrett's epithelium. AB - BACKGROUND/AIMS: The incidence of adenocarcinoma of the oesophagus has increased. Its major risk factor is Barrett's epithelium of which the sine qua non is microscopically diagnosed intestinal metaplasia. Short segment Barrett's epithelium may often be overlooked during routine endoscopy. In routine biopsies taken from normal-appearing mucosa of the distal oesophagus, the reported rates of short segment Barrett's epithelium in the distal oesophagus reached 36%. We compared these rates with the results obtained in a community hospital in Israel. METHODS: Consecutive patients undergoing oesophagogastroduodenoscopy were enrolled. Biopsy specimens taken from cardia, oesophagogastric junction and 2 cm above the oesophagogastric junction were stained with haematoxylin & eosin and Alcian blue. RESULTS: There were 112 study patients (mean age +/- SD 48. 9+/-18.3 years, 51.8% males). Nine (8.04%) patients had intestinal metaplasia (according to specimen from 2 cm above oesophagogastric junction), and symptoms of gastro oesophageal reflux were found in only four (44.4%) of them. Of these nine patients, six (6.66%) had normal-appearing mucosa and three (3.33%) had macroscopic Barrett's epithelium. Alcian blue staining revealed two patients with intestinal metaplasia that haematoxylin & eosin staining had missed. CONCLUSION: We found an 8% prevalence of intestinal metaplasia compared to 18-36% reported in the literature. We also determined that the added advantage of routine biopsy was 5.4%. PMID- 11432510 TI - Evaluation of the association between serum anti-Lewis X antibody and inflammatory infiltration into the gastric mucosa infected with Helicobacter pylori. AB - BACKGROUND: Lipopolysaccharides of Helicobacter pylori have an antigenic structure that mimics Lewis X occurring in gastric mucosa. The pathogenic role of antigenic mimicry in Helicobacter pylori-induced gastritis has been of recent interest. AIM: To examine whether this molecular mimicry affects gastric mucosal inflammation in patients infected with Helicobacter pylori. PATIENTS: A total of 59 patients (mean age 58.0 years, 35 males, 24 females) were studied. METHODS: Serum samples were collected to measure IgG antibodies to Helicobacter pylori and Lewis X. Biopsy specimens were obtained from the antrum and corpus for the grading of gastritis. Correlation coefficients between serum Lewis X antibody titre and histological grades of inflammatory infiltration were determined. RESULTS: There was no significant correlation between anti-Lewis X antibody titre and the grade of mononuclear or polymorphonuclear cell infiltration. High titre of anti-Lewis X antibody was seen only in patients who had increased inflammatory infiltration in the corpus. CONCLUSIONS: Serum anti-Lewis X antibody, possibly induced by Helicobacter pylori infection, does not seem to play a major role in gastric mucosal inflammation in patients with Helicobacter pylori infection. PMID- 11432511 TI - Long-acting octreotide is effective in controlling rebleeding angiodysplasia of the gastrointestinal tract. AB - BACKGROUND: Management of bleeding angiodysplasia of the gastrointestinal tract is often a major clinical problem. Lesions are frequently multiple, not detectable or missed during conventional endoscopy and patients are sometimes at high risk for complications because of advanced age and serious concomitant disorders. AIMS: To determine the efficacy of a new formulation of somatostatin analogue (octreotide long-acting) in management of recurrent bleeding angiodysplasia in patients resistant to endoscopic treatment and not suitable for surgery. PATIENTS AND METHODS: Three patients with recurrent bleeding angiodysplasia of gastrointestinal tract were treated with long-acting octreotide administered intramuscularly 20 mg monthly to each individual. The number of admissions for acute bleeding, hospital stay and number of blood units transfused before and after treatment (followup: 15-17 months) were regularly monitored. RESULTS: In each patient, a relevant decrease in number of hospital admissions, duration of hospital stay, number of administered blood units was seen and mean haemoglobin values significantly increased in all of them after introducing long acting octreotide therapy. CONCLUSIONS: This is the first report on use of long acting octreotide in bleeding angiodysplasia of gastrointestinal tract. Data suggest that long-acting octreotide is a safe drug and is successful in controlling recurrent gastrointestinal bleeding due to angiodysplasia in elderly patients not eligible for surgical or endoscopic therapy. PMID- 11432512 TI - Comparison of topically applied 0.2% glyceryl trinitrate ointment, incision and excision in the treatment of perianal thrombosis. AB - BACKGROUND AND AIMS: Topically applied 0.2% glyceryl trinitrate ointment to the anal region, incision and excision were compared in the treatment of perianal thrombosis, in order to establish which method is the best in terms of pain relief, number of recurrences and the appearance of anal skin tags. PATIENTS AND METHODS: A total of 150 patients were randomly divided into three groups of 50, each group being treated by one of the above-mentioned methods. The results of the treatment were evaluated during the first 4 days, after one month and after one year. RESULTS: A statistically significant reduction of pain was achieved by excision if compared with incision or conservative treatment with glyceryl trinitrate (p<0.001), and by conservative treatment with glyceryl trinitrate if compared with incision (p<0.01 on the 4th day of follow-up. Comparison of patients treated by the three different methods did not reveal any significant difference between these methods after one month (p>0.05). After one year the number of recurrences was significantly reduced after excision if compared with incision (p<0.05) or conservative treatment with glyceryl trinitrate (p<0.05). The number of patients without symptoms was significantly greater after excision if compared with incision or conservative treatment with glyceryl trinitrate (p<0.001). The number of patients with anal skin tags was significantly reduced in patients treated by excision if compared with incision (p<0.001) or glyceryl trinitrate treatment (p<0.001). CONCLUSION: Excision is a significantly better method of treatment of perianal thrombosis than incision or topically applied 0.2% glyceryl trinitrate ointment. PMID- 11432513 TI - Avoiding early postoperative complications in liver surgery. A multivariate analysis of 254 patients consecutively observed. AB - BACKGROUND: The number of hepatic resections for benign and malignant lesions has constantly increased over the past 20 years, as a consequence, surgical experience acquired over the past few years has decreased post-operative morbidity and mortality rates. AIMS: Analysing the relation between potential preoperative risk factors and the occurrence of severe post-operative complications, an attempt is made to identify the variables determining surgical risk in elective hepatic surgery both in normal and cirrhotic liver. PATIENTS AND METHODS: The hospital records of 254 patients who underwent elective liver surgical procedures for hepatic lesions in our department, between 1984 and 1999, were reviewed. The following variables were entered into univariate and multivariate analysis: age, sex, nature of liver lesion (benign or malignant), presence of cirrhosis or cholestasis, synchronous resection of other organs, disorders of blood coagulation, intraoperative blood requirement, the extent of surgical procedures and Pringle's manoeuvre. RESULTS AND CONCLUSIONS: The multivariate analysis of the 254 surgical operations on the liver indicates that the most powerful independent predictors favouring a serious adverse effect includes intra-operative blood transfusions, advanced age and cirrhosis. Scrupulous preoperative clinical evaluation and expert surgical skills minimize intra-operative bleeding and proved to be the most significant factors influencing morbidity and mortality rates. PMID- 11432514 TI - Human leucocyte interferon-alpha in the treatment of chronic hepatitis C. AB - AIM: To assess the efficacy of different schedules of human leucocyte interferon alpha in chronic hepatitis C. PATIENTS AND METHODS: A total of 213 naive patients with chronic hepatitis C were treated with 4 different schedules of human leucocyte interferon alpha. Sustained response was defined as persistently normal alanine amino transferase values with negative serum hepatitis C virus-RNA up to 12 months after therapy withdrawal. RESULTS: Rates of sustained response were 16% with 3 MU tiw for 6 months, 33% with 6 MU tiw for 5 months after a priming dose of 9 MU tiw for a month, 32% with 3 MU tiw for 12 months and 20% with 3 MU daily for 6 months. The major factors affecting the response rate were age and the hepatitis C virus genotype, as a sustained response was significantly higher in patients under 45 years and infected by hepatitis C virus types other than hepatitis C virus-1. Treatment was well tolerated and side-effects and drop-out events were similar to those described with other types of alpha-interferons. CONCLUSIONS: Human leucocyte interferon alpha appears to be equivalent to recombinant interferon-alpha in the treatment of chronic hepatitis C. PMID- 11432515 TI - Insulin resistance in non-alcoholic steatohepatitis. AB - BACKGROUND: Non-alcoholic steatohepatitis is a chronic liver disease that is capable of progressing to end-stage liver disease, but generally has a benign course. Obesity, non-insulin-dependent diabetes mellitus and hyperlipidaemia are the most common associations of the disease. AIMS: To investigate the insulin resistance in patients with non-alcoholic steatohepatitis who have no other causes of insulin resistance such as obesity, diabetes mellitus, and hyperlipidaemia. PATIENTS: Thirteen patients (7 male, 6 female) with non alcoholic steatohepatitis and 12 (6 male, 6 female) healthy volunteers. METHODS: All patients and healthy volunteers were submitted to biochemical tests and hyperinsulinaemic euglycaemic insulin clamp technique. RESULTS: Basal insulin levels and C-peptide levels were significantly higher in non-alcoholic steatohepatitis group than in controls (p<0.001 and p<0.001, respectively). Hyperinsulinaemic euglycaemic insulin clamp technique revealed lower glucose utilization in the non-alcoholic steatohepatitis group and the difference was statistically significant (p<0.001). CONCLUSIONS: Our study revealed marked hyperinsulinaemia and insulin resistance in patients with non-alcoholic steatohepatitis. Hyperinsulinaemia and insulin resistance may contribute to pathogenesis of nonalcoholic steatohepatitis. PMID- 11432516 TI - "Hepatitic flare", asthenia, peripheral polyneuropathy and diffuse liver steatosis in a hepatitis C virus asymptomatic chronic carrier. AB - In July 2000, a 62-year-old female, with a ten-year history of chronic hepatitis C virus infection and persistently normal aspartate amino-transferase and alanine aminotransferase levels, presented with asthenia, weight loss, peripheral polyneuropathy and increased levels of aspartate aminotransferase (8 times upper normal limit), alanine aminotransferase (10 times upper normal limit) and gamma glutamyl-transferase (6 times upper normal limit). The ultrasound findings were consistent with massive liver steatosis. The patient had been previously diagnosed elsewhere as having hepatitis C virus-related "hepatitic flare" with neurological involvement related to concomitant mixed type-III cryoglobulinaemia. However intense exposure to trichloroethylene since April 2000 was revealed and liver histology was fully consistent with non-alcoholic steatohepatitis. The pathogenetic role of the solvent was definitely supported by the complete clinical and biochemical remission within six months of trichloroethylene withdrawal. PMID- 11432517 TI - Seizures in an interferon-treated child. AB - Interferon-treated patients can present seizures, which in most paediatric cases are related to fever. The case of chronic hepatitis C is described in which Interferon probably disclosed a latent epilepsy. The hypothesis is advanced that seizures can be provoked by Interferon therapy in subjects with a low convulsant threshold and, in those cases in which it cannot be substituted with another drug, antiepileptic therapy should be started after the first episode. PMID- 11432518 TI - Clinical course of pregnant women with chronic hepatitis C virus infection and risk of mother-to-child hepatitis C virus transmission. AB - As far as concerns chronic hepatitis C virus infection in pregnant women, different points remain to be elucidated, such as the clinical course, the rate of mother-to-child hepatitis C virus transmission and, in particular, its route and the possible risk factors. This review aimed to analyse current data on the prevalence of chronic hepatitis C virus infection in pregnant women and its relationship with risk factors, the rate of mother-to-child hepatitis C virus transmission and the factors possibly involved, particularly the maternal hepatitis C virus viral load and the human immunodeficiency virus coinfection, and the type of delivery and feeding. Finally, the appropriate timing for HCV-RNA testing in newborns has been reviewed. PMID- 11432519 TI - Pathology of colorectal cancer. AB - The earliest phases of colorectal tumourigenesis initiate in the normal mucosa, with a generalised disorder of cell replication, and with the appearance of clusters of enlarged crypts (aberrant crypts) showing proliferative, biochemical and biomolecular abnormalities. The large majority of colorectal malignancies develop from adenomatous polyps. These can be defined as well demarcated masses of epithelial dysplasia, with uncontrolled crypt cell division. An adenoma can be considered malignant when neoplastic cells pass through the muscularis mucosae and infiltrate the submucosa. Definitions like "carcinoma in situ" or "intramucosal carcinoma" should be abandoned, since they lead to confusion. Although several lines of evidence indicate that carcinomas usually originate from pre-existing adenomas, this does not imply that all polyps undergo malignant changes, and does not exclude "de novo" carcinogenesis. Besides adenomas, other types of polypoid lesions include hyperplastic polyps (showing elongated crypts often with cystic dilatation), serrated adenomas (with a serrated glandular pattern], flat adenomas (flat lesions which are difficult to detect in routine lower endoscopy, but may possess malignant potential), hamartomatous polyps (which show a complex branching pattern of smooth muscle supporting normal lamina propria and glands), and inflammatory polyps. Colorectal carcinomas are one of the most frequent neoplasms in Western society; the macroscopic appearance of these lesions may be that of a polypoid vegetating mass or of a flat infiltrating lesion. Most of these tumours are adenocarcinomas (96%), that, in some cases, show a mucinous component. More rare malignancies of the large bowel include signet-ring cell carcinoma, squamous carcinoma, undifferentiated neoplasms and medullary type adenocarcinoma (solid carcinoma with minimal glandular differentiation or slight cellular pleomorphism). Colorectal carcinoma can be graded into well, moderately and poorly differentiated lesions; there is little evidence, however, that grading may be of help in evaluating prognosis of affected patients. In conclusion, colorectal tumours cover a wide range of premalignant and malignant lesions, many of which can easily be removed at endoscopy. It follows that colorectal neoplasms might be prevented by interfering with the various steps of carcinogenesis, which begins with uncontrolled epithelial cell replication, continues with the formation of adenomas of various dimensions, and eventually evolves into malignancy. PMID- 11432520 TI - Effective antiviral treatment for hepatitis C virus-related chronic active hepatitis in haemophiliac patients. PMID- 11432521 TI - Diagnostic yield of positive immunochemical occult-blood test by digital rectal examination: comparison between subjects with and without haemorrhoids. PMID- 11432522 TI - Helicobacter pylori infection in diabetic patients with dyspepsia in Uganda. PMID- 11432523 TI - Autoimmune serology of primary sclerosing cholangitis. PMID- 11432524 TI - Murray Valley encephalitis virus surveillance and control initiatives in Australia. National Arbovirus Advisory Committee of the Communicable Diseases Network Australia. AB - Mechanisms for monitoring Murray Valley encephalitis (MVE) virus activity include surveillance of human cases, surveillance for activity in sentinel animals, monitoring of mosquito vectors and monitoring of weather conditions. The monitoring of human cases is only one possible trigger for public health action and the additional surveillance systems are used in concert to signal the risk of human disease, often before the appearance of human cases. Mosquito vector surveillance includes mosquito trapping for speciation and enumeration of mosquitoes to monitor population sizes and relative composition. Virus isolation from mosquitoes can also be undertaken. Monitoring of weather conditions and vector surveillance determines whether there is a potential for MVE activity to occur. Virus isolation from trapped mosquitoes is necessary to define whether MVE is actually present, but is difficult to deliver in a timely fashion in some jurisdictions. Monitoring of sentinel animals indicates whether MVE transmission to vertebrates is actually occurring. Meteorological surveillance can assist in the prediction of potential MVE virus activity by signalling conditions that have been associated with outbreaks of Murray Valley encephalitis in humans in the past. Predictive models of MVE virus activity for south-eastern Australia have been developed, but due to the infrequency of outbreaks, are yet to be demonstrated as useful for the forecasting of major outbreaks. Surveillance mechanisms vary across the jurisdictions. Surveillance of human disease occurs in all States and Territories by reporting of cases to health authorities. Sentinel flocks of chickens are maintained in 4 jurisdictions (Western Australia, the Northern Territory, Victoria and New South Wales) with collaborations between Western Australia and the Northern Territory. Mosquito monitoring complements the surveillance of sentinel animals in these jurisdictions. In addition, other mosquito monitoring programs exist in other States (including South Australia and Queensland). Public health control measures may include advice to the general public and mosquito management programs to reduce the numbers of both mosquito larvae and adult vectors. Strategic plans for public health action in the event of MVE virus activity are currently developed or being developed in New South Wales, the Northern Territory, South Australia, Western Australia and Victoria. A southern tri-State agreement exists between health departments of New South Wales, Victoria and South Australia and the Commonwealth Department of Health and Aged Care. All partners have agreed to co-operate and provide assistance in predicting and combatting outbreaks of mosquito-borne disease in south-eastern Australia. The newly formed National Arbovirus Advisory Committee is a working party providing advice to the Communicable Diseases Network Australia on arbovirus surveillance and control. Recommendations for further enhancement of national surveillance for Murray Valley encephalitis are described. PMID- 11432525 TI - Murray Valley encephalitis in Mt Isa, north Queensland. Tropical Public Health Unit Network, Queensland Health. PMID- 11432526 TI - Central Australian MVE update, 2001. PMID- 11432527 TI - Australian encephalitis: sentinel chicken surveillance programme. PMID- 11432528 TI - Report of the Australian National Polio Reference Laboratory, 1 July to 31 December 2000. AB - The Australian National Polio Reference Laboratory at the Victorian Infectious Diseases Reference Laboratory (VIDRL) is responsible for processing and testing samples for poliovirus from all Australian patients with acute flaccid paralysis and for identifying and characterising polioviruses recovered from untyped enteroviruses submitted from Australian laboratories. From 1 July to 31 December 2000, a total of 12 specimens from 7 patients with AFP were referred to the NPRL. Poliovirus type 3 Sabin-like was isolated from samples from 2 patients with suspected vaccine- associated paralytic poliomyelitis. No viruses were isolated from samples from the remaining 5 patients. Since 1995 a total of 1,325 isolates have been referred for testing from laboratories throughout Australia. Seven hundred (53%) were confirmed as Sabin vaccine-like polioviruses, 542 (41%) were non-polio enteroviruses and 82 (6%) yielded no virus or viruses other than enteroviruses. At Kyoto, Japan in October 2000, the Western Pacific Region of the World Health Organization was declared wild polio-free. This represents a significant step towards the global eradication of poliovirus with one quarter of the world's population free of endemic infections from wild poliovirus. Surveillance of AFP and containment of wild polioviruses has been coordinated at the VIDRL. Since February 2000, Australia has been developing and implementing a plan for the containment of wild poliovirus stocks and potentially infectious materials. PMID- 11432529 TI - Annual report of the Australian Gonococcal Surveillance Programme, 2000. AB - The Australian Gonococcal Surveillance Programme (AGSP) monitors the antibiotic susceptibility of Neisseria gonorrhoeae isolated in all States and Territories. In 2000 the in vitro susceptibility of 3,468 isolates of gonococci was determined by standardised methods. Antibiotic susceptibility patterns varied considerably between regions. Resistance to the penicillins was highest in larger urban centres and warrants close attention in those rural centres where treatment with the penicillins continues. Quinolone resistance in gonococci became more widespread in Australia in 2000. Endemic cycles of transmission of quinolone resistant gonococci (QRNG) in homosexually active men continued in Victoria but declined in New South Wales. Heterosexual endemic transmission of QRNG increased substantially in New South Wales and the proportion of all gonococci represented by QRNG also increased markedly in Queensland and Western and South Australia. All isolates remained sensitive to spectinomycin, but a small number of isolates in a number of jurisdictions showed some decreased susceptibility to ceftriaxone. Strains examined in South Australia, New South Wales and Victoria were predominantly from male patients and rectal and pharyngeal isolates were common. In other centres the male to female ratio of cases was lower, and most isolates were from the genital tract. PMID- 11432530 TI - Investigation and control of a cluster of cases of Legionnaires disease in western Sydney. AB - Three cases of Legionella pneumophila infection were identified in Sydney's west in November 1998. Epidemiological investigations identified an association with one workplace. Environmental sampling revealed that the cooling towers in the workplace, and at 2 other premises within a 1 km radius of the workplace, were positive for L. pneumophila serogroup 1 (LP1) which was indistinguishable from clinical isolates of 2 of the cases on DNA fingerprinting. Our report highlights limitations of the current control program for Legionella in cooling towers, including the finding of unregistered cooling towers, cooling towers positive for LP1 despite satisfactory results on inspection, and cooling towers potentially linked to disease with counts of LP1 below the current protocol requirements for immediate decontamination. PMID- 11432531 TI - Prevalence of tattooing and body piercing in the Australian community. AB - Tattooing and body piercing are now worldwide fashion crazes. The health risks associated with these procedures are as yet unclear. This article examines the prevalence of body decoration and the associated health risks within the Australian population using a random sample survey of individuals aged 14 years and over, collected between June and September 1998 (n = 10,030). The results show that one in 10 people have had a tattoo at some point in their lives and 8 per cent some form of body piercing, excluding ear piercing. Men are more likely than women to report tattooing, while females are more likely to report body and ear piercing. Some 10 per cent of respondents report drinking alcohol or using other drugs when the procedures were undertaken. The prevalence of tattooing and body piercing is considerably higher among injecting drug users. Although the rates of transmission of bloodborne disease due to body decoration are believed to be low, the strong association with youth and with injecting drug use suggests considerable potential for transmission. PMID- 11432532 TI - An outbreak of Salmonella Typhimurium phage type 29 linked to a noodle restaurant in South Australia. PMID- 11432534 TI - Communicable Diseases Surveillance. PMID- 11432533 TI - An outbreak of Salmonella Typhimurium phage type 126 linked to a cake shop in South Australia. PMID- 11432535 TI - Concentration-controlled or effect-controlled trials: useful alternatives to conventional dose-controlled trials? AB - Historically, dose-finding trials have been confirmatory in nature despite the fact that these trials represent an important and essential 'learning' phase in the drug development process. About 10 years ago 2 alternatives to the randomised dose-controlled trial (RDCT) were proposed as being more informative trial types. Controlling systemic drug exposure in order to improve efficiency of a trial forms the basis for the suggestion of a randomised concentration-controlled trial (RCCT). For the common instance where pharmacodynamic variability is larger than pharmacokinetic variability, the randomised effect-controlled trial (RECT), where patients are randomised to the effect of interest was suggested as even more informative. A survey of the literature shows that the RCCT has been sparsely applied and RECT not at all. For RCCT, the practical complications of carrying out the study seldom makes it the study type of choice. For RECT, the limited number of suitable situations for its application and the fact that the same effect is used for randomisation and analysis may explain the lack of applications. As a somewhat more favourable trial type, we suggest the randomised biomarker-controlled trial (RBCT), where patients are randomised to a certain value or range of a biomarker whereas the analysis is performed on another, clinically more relevant, effect. Although the RBCT has some attractive features, for example contributing to validation of a biomarker as a surrogate for clinical outcome, it is unlikely to be extensively used. Instead, the main shift from confirming to learning in dose-finding trials is coming from the incorporation of well-known learning components into the RDCT (e.g. sparse concentration measurements combined with population pharmacokinetic-pharmacodynamic, biomarker measurements and analysis of effect measures throughout the entire trial period). PMID- 11432536 TI - Comparative pharmacokinetics and pharmacodynamics of the rifamycin antibacterials. AB - The rifamycin antibacterials, rifampicin (rifampin), rifabutin and rifapentine, are uniquely potent in the treatment of patients with tuberculosis and chronic staphylococcal infections. Absorption is variably affected by food; the maximal concentration of rifampicin is decreased by food, whereas rifapentine absorption is increased in the presence of food. The rifamycins are well-known inducers of enzyme systems involved in the metabolism of many drugs, most notably those metabolised by cytochrome P450 (CYP) 3A. The relative potency of the rifamycins as CYP3A inducers is rifampin > rifapentine > rifabutin; rifabutin is also a CYP3A substrate. The antituberculosis activity of rifampicin is decreased by a modest dose reduction from 600 to 450mg. This somewhat surprising finding may be due to the binding of rifampicin to serum proteins, limiting free, active concentrations of the drug. However, increasing the administration interval (after the first 2 to 8 weeks of therapy) has little effect on the sterilising activity of rifampicin, suggesting that relatively brief exposures to a critical concentration of rifampicin are sufficient to kill intermittently metabolising mycobacterial populations. The high protein binding of rifapentine (97%) may explain the suboptimal efficacy of the currently recommended dose of this drug. The toxicity of rifampicin is related to dose and administration interval, with increasing rates of presumed hypersensitivity with higher doses combined with administration frequency of once weekly or less. Rifabutin toxicity is related to dose and concomitant use of CYP3A inhibitors. The rifamycins illustrate the complexity of predicting the pharmacodynamics of treatment of an intracellular pathogen with the capacity for dormancy. PMID- 11432537 TI - Pharmacokinetic interactions of antimalarial agents. AB - Combination of antimalarial agents has been introduced as a response to widespread drug resistance. The higher number of mutations required to express complete resistance against combinations may retard the further development of resistance. Combination of drugs, especially with the artemisinin drugs, may also offer complete and rapid eradication of the parasite load in symptomatic patients and thus reduce the chance of survival of resistant strains. The advantages of combination therapy should be balanced against the increased chance of drug interactions. During the last decade, much of the pharmacokinetics and metabolic pathways of antimalarial drugs have been elucidated, including the role of the cytochrome P450 (CYP) enzyme complex. Change in protein binding is not a significant cause of interactions between antimalarial agents. CYP3A4 and CYP2C19 are frequently involved in the metabolism of antimalarial agents. Quinidine is a potent inhibitor of CYP2D6, but it appears that this enzyme does not mediate the metabolism of any other antimalarial agent. The new combinations proguanil atovaquone and chlorproguanil-dapsone do not show significant interactions. CYP2B6 and CYP3A4 are involved in the metabolism of artemisinin and derivatives, but further studies may reveal involvement of more enzymes. Artemisinin may induce CYP2C19. Several artemisinin drugs suffer from auto-induction of the first pass effect, resulting in a decline of bioavailability after repeated doses. The mechanism of this effect is not yet clear, but induction by other agents cannot be excluded. The combination of artemisinin drugs with mefloquine and the fixed combination artemether-lumefantrine have been studied widely, and no significant drug interactions have been found. The artemisinin drugs will be used at an increasing rate, particularly in combination with other agents. Although clinical studies have so far not shown any significant interactions, drug interactions should be given appropriate attention when other combinations are used. PMID- 11432538 TI - Application of a gamma model of absorption to oral cyclosporin. AB - BACKGROUND: Some drugs, such as cyclosporin, exhibit flat and delayed absorption profiles, with a correlation between the delay and the peak width. Such profiles can be described by an absorption model in which the absorption rate is derived from a gamma distribution (of which the classical first-order absorption model is a special case). OBJECTIVE: To develop a model for the pharmacokinetics of extravascular administration of cyclosporin and apply it to a study of the pharmacokinetics of cyclosporin microemulsion in stable renal transplant recipients. PATIENTS AND PARTICIPANTS: 21 renal transplant patients receiving oral cyclosporin microemulsion 75 to 175 mg twice daily. METHODS: The equation of the plasma concentration-time curve after oral administration was expressed as a convolution product between the absorption rate and a multi-exponential impulse response. The convolution integral was computed analytically and expressed in terms of the incomplete gamma function. Cyclosporin was assayed by liquid chromatography/mass spectrophotometry. The model was fitted by nonlinear regression, using a specially developed program. RESULTS: The gamma model yielded a good fit in all of the 21 patients studied. Attempts to fit the same data by a classical exponential with lag-time model failed in most patients. CONCLUSIONS: This model could simplify the Bayesian monitoring of cyclosporin therapy. PMID- 11432539 TI - Pharmacokinetic-pharmacodynamic relationship of levodopa with and without tolcapone in patients with Parkinson's disease. AB - OBJECTIVE: To investigate the effect of administration of the catechol Omethyltransferase (COMT) inhibitor tolcapone on the concentration-effect relationship of levodopa in patients with advanced Parkinson's disease and on-off fluctuations. DESIGN: Nonblind single-group 2-period pharmacokinetic pharmacodynamic study. PATIENTS AND PARTICIPANTS: 12 patients, mean age 59 years, with idiopathic Parkinson's disease and response fluctuations. METHODS: The pharmacokinetics [plasma concentrations of levodopa and 3-O-methyldopa (3-OMD)] and motor effects [global score of the Columbia University Rating Scale (CURSsigma)] of levodopa (plus the peripheral decarboxylase inhibitor benserazide 1:4) were determined for 4 consecutive dosage intervals (4 hours each, starting at 8.00am) in 12 patients before (day 1) and during (day 8) coadministration of tolcapone 100 mg 3 times daily for 7 days. RESULTS: Under tolcapone, exposure to levodopa [area under the plasma concentration-time for the dosage interval (AUCt)] observed for the separate doses increased by 1.6- to 2.2-fold, and peak plasma drug concentrations (Cmax) increased by 1.1 - to 2.1 -fold. 3-OMD concentrations at day 8 were reduced to about 20% of the values at day 1. At baseline (day 1, before the first levodopa dose), CURSsigma averaged 40 +/- 10 points. After the first levodopa dose. CURSsigma declined to 20 +/- 9 points. At day 8. the predose CURSsigma decreased to a final score of 31 +/-13 points, and the maximal decline after the first levodopa dose was to a final score of 16 +/- 8 points. Population analysis (NONMEM) of the concentration-effect relationship of levodopa according to a sigmoidal Emax model and over all dosage intervals did not show differences in levodopa responsiveness with or without tolcapone. The population mean of the 50% effective concentration (EC50) of levodopa was 1350 microg/L with an standard error of the population parameter estimate of 18%: adding tolcapone treatment as a covariate did not significantly change the population fit. Circadian influences on levodopa respon- siveness were not evaluable by the NONMEM model due to overparametrisation, but visual inspection of plotted data did not suggest differences in the concentration-effect relationship between the 4 consecutive dosage intervals on days 1 and 8. CONCLUSIONS: The gain in clinical improvement with levodopa under tolcapone can be fully explained by tolcapone-induced changes of peripheral levodopa pharmacokinetics. We suggest that this interaction study, performed in patients and using clinical data, excludes any central effects of tolcapone or any inhibiting effect of 3-OMD on levodopa permeation through the blood-brain barrier, which otherwise would have led to a decrease in the EC50 of levodopa. PMID- 11432540 TI - An analysis of soil arsenic records of decision. AB - In 1986 the US EPA created the National Priority List (NPL) that now comprises in excess of 2,000 sites nationwide, with arsenic the second most common inorganic constituent. A survey of 69 Records of Decision (RODs) written between 1985 and 1998 for which arsenic was a major driver found that 84% of cleanup goals were risk-driven and 16% were background-driven, with a wide range of soil-arsenic cleanup standards for 10(-6) residential risk goals (2-305 mg/kg). In comparison, the range of background-based cleanup goals was much narrower (8-21 mg/kg). ROD soil arsenic concentrations exhibit no statistically significanttemporal trend, but on a geographic basis, EPA Regions 6, 8, 9, and 10 had some of the higher decisions. The risk assessment process is important in defining cleanup goals; however routine use of site-specific variables (i.e., bioavailability, realistic tenure in both residential and occupational settings, natural attenuation of arsenic in groundwater, etc.) is necessary to ensure an accurate assessment of potential site risks and to preclude over-remediation that may result from the use of default risk variables. PMID- 11432541 TI - Generation of enterococci bacteria in a coastal saltwater marsh and its impact on surf zone water quality. AB - Elevated levels of enterococci bacteria, an indicator of fecal pollution, are routinely detected in the surf zone at Huntington State and City Beaches in southern California. A multidisciplinary study was carried out to identify sources of enterococci bacteria landward of the coastline. We find that enterococci bacteria are present at high concentrations in urban runoff, bird feces, marsh sediments, and on marine vegetation. Surprisingly, urban runoff appears to have relatively little impact on surf zone water quality because of the long time required for this water to travel from its source to the ocean. On the other hand, enterococci bacteria generated in a tidal saltwater marsh located near the beach significantly impact surf zone water quality. This study identifies a potential tradeoff between restoring coastal wetlands and protecting beach water quality and calls into question the use of ocean bathing water standards based on enterococci at locations near coastal wetlands. PMID- 11432542 TI - A comparison of PAH, PCB, and pesticide concentrations in air at two rural sites on Lake Superior. AB - Atmospheric concentrations of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and pesticides were compared at Brule River and Eagle Harbor, two rural sites on Lake Superior that are part of the Integrated Atmospheric Deposition Network (IADN). Brule River lies 40 km southwest of Duluth, MN, a small industrial city, and Eagle Harbor is in Michigan's upper peninsula, 400 km east of Brule River. Pesticide and PCB concentrations were similar at both sites. Day-by-day regression analyses of the data showed that PAH concentrations, an indication of urban contamination, were significantly higher at Brule Riverthan at Eagle Harbor. Concentration ranges for all compounds at both sites were well within global background levels, despite the differences observed between the two sites. Clearly, pollution from Duluth is influencing PAH concentrations at Brule River more than at Eagle Harbor. PMID- 11432543 TI - Identification of the isoflavonoid genistein in bleached kraft mill effluent. AB - Plants synthesize many phytochemicals, including flavonoids, which may be present in the heartwood of trees used in the pulp and paper industry. Extracts were prepared from wood pulp and mill effluent collected from a bleached kraft mill in Ontario, Canada, and these extracts were subfractionated by LH-20 gel filtration chromatography and analyzed by liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS) and LC-ESI-tandem mass spectrometry (LC-ESI-MS-MS). Initial LC-MS analysis in negative ion mode was conducted by monitoring ions corresponding to the deprotonated molecular ions of a range of flavonoid compounds. The presence of chromatographic peaks generated by selected ion monitoring (SIM) in samples of both mill effluent and wood pulp encouraged further analysis. Of the compounds highlighted by LC-ESI-MS-SIM analysis, the isoflavonoid genistein was positively identified by LC-ESI-MS-MS. Genistein was quantified at a concentration of 30.0 microg/kg in air-dried wood pulp and concentrations of 13.1 microg/L and 10.5 microg/L in untreated and treated (final) effluent, respectively. Genistein is a known endocrine disruptorsubstance and,therefore, could contribute to the alterations in sex steroid levels and reduced reproductive capacity observed in fish captured near the discharges of pulp mills. PMID- 11432544 TI - Distribution and fate of neutral alkylphenol ethoxylate metabolites in a sewage impacted urban estuary. AB - The distribution and fate of neutral metabolites of the alkylphenol ethoxylate (APEO) surfactants in an urbanized estuarine environment were examined utilizing a recently developed, highly sensitive LC-MS method. Results indicated that short ethoxyl-chain APEOs and alkylphenols (APs) were present in surficial sediments throughout the estuary at concentrations roughly correlated to the organic carbon content of the sediment and that the APEO mixture was dominated by nonylphenol ethoxylate (NPEOs) metabolites (0.05-30 microg/g), with lesser amounts of octylphenol ethoxylate metabolites (OPEOs)(<0.005-0.09 microg/ g) and halogenated nonylphenols (<0.001-0.03 microg/g). NPEO metabolites in surface water (0.22-1.05 microg/L) were also present at higher concentrations than OPEO metabolites (0.007 0.040 microg/L). APEO metabolite concentrations in both sediment and water showed a strong correlation with conventional sewage tracers, affirming a wastewater source of these contaminants. APEO distributions in surface waters within the estuary could be explained by a combination of post-discharge degradation and mixing with a seawater end-member enriched in OPEO metabolites. Measured in situ Koc values of APEO metabolites were comparable to previously reported values derived from field experiments but higher than Kow and Koc values derived from laboratory experiments. Results from the present work indicate that the fate of APEO metabolites entering the estuarine environment through discharge of wastewater is directed primarily by scavenging onto particles and subsequent burial in sediments, degradation during residence in the water column, and transport out of the estuary through advective and dispersive processes. PMID- 11432545 TI - Study on the abundance of CFCs varying with the latitude at the bottom of the troposphere in the southern hemisphere. AB - As the main ozone depleting compounds, the abundance of CFCs in the troposphere and stratosphere has been focused on all the while. However, there are few reports concerning on the CFCs' latitudinal dependence. Relying on the longterm monitoring that has been performed in Shanghai from 1997 and Cooperating with the Chinese Third Antarctic Inland Ice-sheet Traversal Party, we did intensive sampling in the southern hemisphere from the beginning of November to the end of December in 1998 and then got the corresponding concentrations of CFC-12, CFC-11, and CFC-113 according to the latitude. The data were studied inductively in accordance with the geographical location and meteorological condition of every sampling point. In summary, the average concentrations of CFCs (refer to CFC-11, CFC-12, and CFC-113) in equatorial area are the highest in the southern hemisphere; however, the concentrations in prevailing west-wind belt and polar east-wind belt are much lower and at the same levels, which should be a perfect implication of the mean background in the southern hemisphere. Being related to the special polar circumfluence, the concentrations of CFCs on the glacier reveal a special rise. Compared with the mean background of the southern hemisphere that is represented by the data of the prevailing west-wind belt and Polar east-wind belt, the concentrations in Shanghai bear the same level. PMID- 11432546 TI - Refuse incinerator particulate emissions and combustion residues for New York City during the 20th century. AB - Refuse incineration data for New York City (NYC) have been compiled as a function of time during the 20th century to assess the historical significance of this pollutant source in a densely populated area. Thirty-two municipal and 17,000 apartment house refuse incinerators were identified. Approximately 1.1 x 10(8) t of refuse (wet weight) were combusted in NYC incinerators between 1908 and 1993, producing 3.4 x 10(7) t(dryweight) of combustion residue disposed in local landfills. Refuse incinerators were operated for most of this period without air pollution control and emitted 1.0 x 10(6) t of particles (a total of 120 mg for each cm2 of land in NYC). Incinerator particle emission (PE) rates per unit area of land were highest in Manhattan (equivalent total deposition of 530 mg cm(-2)). Incinerator PE exceeded 1.2 x 10(4) t yr(-1) between 1930 and 1975, with maximum emission rates (>2.2 x 10(4) t yr(-1)) in the late 1930s and 1960s. These and other factors support the conclusion that refuse incineration without air pollution control was an important source of airborne, respirable pollutants in NYC for many decades during the 20th century. Rates of particle emissions from Manhattan incinerators estimated here correlate stronglywith Pb accumulation rates as a function of depth (time) in Central Park Lake sediments, consistent with refuse incineration emitting large amounts of atmospheric lead in NYC for many decades afterthe 1920s. PMID- 11432547 TI - Enantiomeric composition of chiral polychlorinated biphenyl atropisomers in aquatic and riparian biota. AB - The enantiomeric composition of polychlorinated biphenyl (PCB) atropisomers was measured in river and riparian biota (fish, bivalves, crayfish, water snakes, barn swallows) from selected sites throughout the United States by using chiral gas chromatography/mass spectrometry. Nonracemic enantiomeric fractions (EFs) were observed for PCBs 91, 95, 136, and 149 for aquatic and riparian biota from Lake Hartwell, SC, a reservoir heavily contaminated with PCBs, and for these congeners and PCBs 132, 174, 176, and 183 in river fish and bivalves nationwide. Fish and bivalves showed marked differences in EFs as compared to sedimentfound atthe same sampling sites, thus suggesting that PCBs are bioprocessed in biota in a different manner from those found in sediment (e.g., reductive dechlorination). Species-dependent patterns in PCB EFs were observed, which suggest differences in the ability of different species to bioprocess PCBs enantioselectively, most likely by metabolism. The presence of nonracemic PCBs in fish and bivalves suggests greater metabolic degradation of PCBs in these organisms than indicated from previous achiral studies and underscores the powerful potential of chiral analysis as a tracer of environmental bioprocesses. PMID- 11432548 TI - Formation and transport of the sulfonic acid metabolites of alachlor and metolachlor in soil. AB - Alachlor and metolachlor are dechlorinated and transformed into their corresponding ethane sulfonic acid (ESA) metabolites in soil. In a field disappearance study, it was shown that alachlor ESA was formed at a faster rate and at concentrations 2-4 times higher than metolachlor ESA, conforming with the observed longer disappearance half-life of metolachlor (15.5 d) in the field as compared to alachlor (8 d). Runoff data also showed higher concentrations of alachlor ESA as compared to metolachlor ESA, even though they were applied at the same levels. Data from soil cores showed transport of the ESA compounds in soil to as far down as 75-90 cm belowthe surface, at concentrations ranging from less than 0.5 microg/L to about 50 microg/L. In contrast, no parent herbicide was detected at these depths. This observation correlates with the higher log Koc values for alachlor (3.33) and metolachlor (3.01) relative to their corresponding ESA metabolites, alachlor ESA (2.26), and metolachlor ESA (2.29). PMID- 11432549 TI - Reduction of the carbamate pesticides oxamyl and methomyl by dissolved FeII and CuI. AB - The degradation of two oxime carbamate pesticides, oxamyl and methomyl, was investigated in anoxic solutions containing various metal ions and reducing agents. In reagent-free solutions, these carbamates degrade slowly via base catalyzed elimination. Rates of carbamate degradation are accelerated by Fe(II), Cu(I), and Cu(II), but not by several other metal ions and reducing agents. In the presence of Fe(II), carbamates undergo a net two-electron reduction that is coupled to the sequential one-electron oxidation of two Fe(II) ions. The observed products are a substituted nitrile, methanethiol, and methylamine. A radical intermediate is inferred by polymerization of the radical scavenger acrylonitrile. Redox kinetics (i) vary with carbamate identity, (ii) exhibit first-order dependence on both Fe(II) and carbamate concentration, (iii) are relatively independent of pH, (iv) follow Arrhenius temperature dependence, and (v) are only indirectly influenced by the presence of O2. Coordinatively saturated Fe(II) complexes (Fe(II)EDTA2- and Fe(II)(CN)6(4-)) react with oxamyl at rates equal to and greater than hexaquo Fe(II), respectively, indicating that an inner-sphere Fe(II)-carbamate coordination complex is not required for electron transfer. Experimental results indicate that Cu(I) reduces the carbamates by the same mechanism as Fe(II) but at much higher rates. In contrast, Cu(II) acts as a catalyst for both elimination and reduction reactions. PMID- 11432550 TI - Sediment-associated reactions of aromatic amines. 1. Elucidation of sorption mechanisms. AB - Sorption of aromatic amines to sediments and soils can occur by both reversible physical processes and irreversible chemical processes. To elucidate the significance of these sorption pathways, the sorption kinetics of aniline and pyridine were studied in resaturated pond sediment. Aniline and pyridine behaved quite differently in the sediment-water systems. The sorption kinetics of pyridine were quite fast, reaching equilibrium within 1-2 h. In contrast, the sorption kinetics of aniline were characterized by a rapid initial loss of aniline from the aqueous phase followed by a much slower rate of disappearance. The rapid initial sorption of aniline upon respiking after an equilibration period of 200 h, and results of sorption kinetic studies as a function of substrate concentration, demonstrated that sorptive sites were not being saturated at the nominal concentration of aniline. Sequential extraction of a sediment treated with 14C-labeled pyridine and aniline suggested that pyridine was bound primarily through a reversible cation-exchange process, whereas aniline sorbed through both cation-exchange and covalent binding processes. At longer reaction periods sorption became increasingly dominated by covalent binding. The reaction kinetics for the slow, irreversible sorption of aniline appeared to be limited by the reactivity and/or availability of covalent binding sites. The initial rate and extent of aniline sorption was pH dependent (sorption increased with decreasing pH). At pH values above the pKa of aniline, sorption kinetics for the slower, irreversible loss of aniline were independent of pH. PMID- 11432551 TI - Assessing the biological potency of binary mixtures of environmental estrogens using vitellogenin induction in juvenile rainbow trout (Oncorhynchus mykiss). AB - Experiments were conducted to assess the in vivo potency of binary mixtures of estrogenic chemicals using plasma vitellogenin (VTG) concentrations in juvenile rainbow trout (Oncorhynchus mykiss) as the endpoint. The estrogenic potencies of estradiol-17beta (E2), 4-tertnonylphenol (NP), and methoxychlor (MXC) were determined following 14 day exposures to the individual chemicals and binary mixtures of these chemicals. E2, NP, and MXC all induced concentration dependent increases in plasma VTG, with lowest observed effect concentrations of 4.7 and 7.9 ng L(-1) for E2, 6.1 and 6.4 microg L(-1) for NP, and 4.4 and 6.5 microg L( 1) for MXC. Concentration-response curves for fixed ratio binary mixtures of E2 and NP (1:1000), E2 and MXC (1:1000), and NP and MXC (1:1) were compared to those obtained for the individual chemicals, using the model of concentration addition. Mixtures of E2 and NP were additive at the concentrations tested, but mixtures of E2 and MXC were less than additive. This suggests that while NP probably acts via the same mechanism as E2 in inducing VTG synthesis, MXC may be acting via a different mechanism(s), possibly as a result of its conversion to HPTE which is an estrogen receptor alpha agonist and an estrogen receptor beta antagonist. It was not possible to determine whether mixtures of MXC and NP were additive using VTG induction, because the toxicity of MXC restricted the effect range forwhich the expected response curve forthe binary mixture could be calculated. The data presented illustrate that the model of concentration addition can accurately predict effects on VTG induction, where we know that both chemicals act via the same mechanism in mediating a vitellogenic response. PMID- 11432552 TI - Kinetic analysis of the bacterial reduction of goethite. AB - The kinetics of dissimilatory reduction of goethite (alpha-FeOOH) was studied in batch cultures of a groundwater bacterium, Shewanella putrefaciens, strain CN32 in pH 7 bicarbonate buffer. The rate and extent of goethite reduction were measured as a function of electron acceptor (goethite) and donor (lactate) concentrations. Increasing goethite concentrations increased both the rate and extent of Fe(III) reduction when cell and lactate concentrations were held constant. However, constant initial reduction rates were observed after normalization to the Fe(II) sorption capacity of FeOOH, suggesting that the bacterial reduction rate was first orderwith respect to surface site concentration. Increasing the lactate concentration also increased the rate and extent of FeOOH reduction. Monod-type kinetic behaviorwas observed with respectto lactate concentration. Fe(II) sorption on FeOOH was well-described by the Langmuir sorption isotherm. However, the Fe(II) sorption capacities hyperbolically decreased with increasing FeOOH concentration (10-100 mM) implying aggregation, while the affinity constant between Fe(II) and goethite was constant (log K approximately equals 3). Evaluation of the end states of the variable FeOOH and lactate experiments when iron reduction ceased indicated a consistent excess in reaction free energy of -22.7 kJ/mol. This value was remarkably close to the minimum value reported for bacteria to mediate a given reaction (-20 kJ/mol). X-ray diffraction (XRD) and scanning electron microscopy (SEM) indicated that siderite (FeCO3) was the only biogenic Fe(II) solid formed upon FeOOH bioreduction. A kinetic biogeochemical model that incorporated Monod kinetics with respect to lactate concentration, first-order kinetics with respectto goethite surface concentration, a Gibbs free energy availability factor, the rates of Fe(II) sorption on goethite and siderite precipitation, and aqueous speciation reactions was applied to the experimental data. Using independently estimated parameters, the developed model successfully described bacterial goethite reduction with variable FeOOH and lactate concentrations. PMID- 11432553 TI - A quantitative relationship that demonstrates mercury methylation rates in marine sediments are based on the community composition and activity of sulfate-reducing bacteria. AB - A quantitative framework was developed which estimates mercury methylation rates (MMR) in sediment cores based on measured sulfate reduction rates (SRR) and the community composition sulfate-reducing bacterial consortia. MMR and SRR as well as group-specific 16S rRNA concentrations (as quantified by probe signal) associated with sulfate-reducing bacteria (SRB) were measured in triplicate cores of saltmarsh sediments. Utilizing previously documented conversion factors in conjunction with field observations of sulfate reduction, MMR were calculated, and the results were compared to experimentally derived measurements of MMR. Using our novel field data collected in saltmarsh sediment where sulfate reduction activity is high, calculated and independently measured MMR results were consistently within an order of magnitude and displayed similar trends with sediment depth. In an estuarine sediment where sulfate reduction activity was low, calculated and observed MMR diverged by greater than an order of magnitude, but again trends with depth were similar. We have expanded the small database generated to date on mercury methylation in sulfur-rich marine sediments. The quantitative frameworkwe have developed further elucidates the coupling of mercury methylation to sulfate reduction by basing calculated rates of mercury methylation on the activity and community composition of sulfate-reducing bacteria. The quantitative framework may also provide a promising alternative to the difficult and hazardous determination of MMR using radiolabeled mercury. PMID- 11432554 TI - Physical and chemical determinants of colloid transport and deposition in water unsaturated sand and Yucca Mountain tuff material. AB - Colloid mobility was determined in a system consisting of quartz sand or crushed Yucca Mountain tuff, simulated groundwater (J-13), and hydrophilic latex particles. Water content (theta) and ionic strength (I; DI water, 0.1x, 1x, 10x groundwater dilution) were manipulated to define limiting conditions for colloid transport atthe Yucca Mountain site. Colloid transport, measured with a centrifuge method at relatively high theta (saturation >36% for sand, >62% for crushed tuff) in DI water, was equivalent to transport at 100% saturation measured with conventional columns. When variables were isolated, increasing I and decreasing theta resulted in a greater extent of colloid deposition; I was more important at higher theta; physical properties were more important at lower theta. I and theta had an interactive effect on colloid deposition whereby synergism was generally observed, especially for simulated groundwater (1x); antagonism was observed at 10x groundwater dilution. At 19% moisture saturation on the crushed tuff, a decreasing rate of colloid deposition was observed. This corresponded to a hydrodynamic condition of 79% immobile water where solute tracers were excluded from a fraction of the pore volume. This suggests that a portion of the favorable sites for deposition were associated with the excluded or immobile water domain and were not accessible to colloids. PMID- 11432555 TI - Relating ion binding by fulvic and humic acids to chemical composition and molecular size. 1. Proton binding. AB - Proton binding by a soil fulvic acid, humic acid, and a set of size fractions of the humic acid was studied as a function of pH and ionic strength by potentiometric titrations. The negative charge of the humic substances resulting from deprotonation of acidic functional groups generally increased with increasing pH and increasing ionic strength. At any given pH and ionic strength, the fulvic acid fraction exhibited much higher negative charge than the humic acid fraction. For the size-fractionated humic acids, negative charge decreased steadily with increasing apparent molecular weight, as determined by size exclusion chromatography. Observed differences in proton binding by the various humic substances corresponded well to differences in functional group composition, which has been extensively characterized in a previous study using a combination of analytical techniques. The proton binding behavior of the humic substances was described very well by the consistent NICA-Donnan model. However, when all adjustable model parameters were determined using a least-squares minimization technique without introducing parameter constraints, the values of some parameters turned out physically and chemically unreasonable. Therefore, we propose to derive some model parameters from chemical characterization results obtained by size exclusion chromatography and solid-state 13C NMR spectroscopy. Using this approach, we obtained excellent descriptions of all titration data, and the model parameter values were more consistent and chemically reasonable. Our results demonstrate that characterization results of humic substances can be used in NICA-Donnan modeling to reduce the number of free fitting parameters without arbitrary constraints and, thereby, obtaining a more reliable database for environmental modeling. PMID- 11432556 TI - Relating ion binding by fulvic and humic acids to chemical composition and molecular size. 2. Metal binding. AB - Binding of Cu(II) and Pb(II) to a soil fulvic acid, humic acid, and two different size fractions of the humic acid was investigated with metal titration experiments at pH 4, 6, and 8. Proton and free metal ion activities in solution were monitored after each titration step using pH and ion selective electrodes (ISE), respectively. The amounts of base required to maintain constant pH conditions were recorded and used to calculate stoichiometric proton-to-metal ion exchange ratios. Despite clear differences in chemical composition and protonation behavior, the fulvic acid and all humic acid fractions exhibited very similar metal binding behavior. Binding of Cu(II) and Pb(II) generally increased with increasing pH and total metal concentration. At low to moderate metal ion concentrations, Cu(II) was bound more strongly to the humic substances than Pb(II). Only at high free metal concentrations, the amounts of metal ions sorbed were higher for Pb(II) than for Cu(II). The molar proton-to-metal ion exchange ratios ranged from 1.0 to 1.8 for Cu(II) and from 0.6 to 1.2 for Pb(II), suggesting that Cu(II) was bound as monodentate and bidentate complexes, while Pb(II) was bound predominantly as monodentate complexes. The metal ion binding data were quantitatively described with the consistent NICA-Donnan model. The best description of an entire multicomponent data set consisting of proton titration, Cu(II), and Pb(II) binding data was achieved when the entire data set was fitted simultaneously. To reduce the number of fitting parameters, results from size exclusion chromatography and solid state 13C NMR spectroscopy were used to estimate two of the NICA-Donnan model parameters. The values of the remaining NICA-Donnan parameters for the humic substances are within a narrow range, suggesting that generalized model parameters may be useful in geochemical modeling involving humic substances. PMID- 11432557 TI - Solvation effect on organic compound interactions in soil organic matter. AB - We examine sorption of pyridine by soil organic matter (SOM) from different organic media including n-hexadecane, acetonitrile, acetone, and n-hexadecane mixtures with either acetonitrile or acetone and compare it with sorption from water. By using an activity-based comparison, we distinguish between solvent assisted and solvent-competitive sorption behavior. Pyridine was selected because it forms strong complexes with phenolic and carboxylic groups, such that site interactions should dominate interactions in SOM. It is anticipated that pyridine sorption will be illustrative of the importance of disrupting strong interactions in a condensed, shrunken SOM phase for many organic compounds. It was generally found that activity-normalized pyridine uptake was assisted by polar solvent molecules rather than suppressed due to competition. An explanation is tendered on the basis of our earlier hypothesis of water-assisted disruption of polar SOM contacts. Certain polar moieties of dry SOM are unavailable for compound sorption due to strong interactions between them. By penetrating SOM structure, solvent molecules (and water) solvate (hydrate) polar moieties creating new sorption sites. Solvent molecules must solvate both moieties of the polar contact, such that the driving force for solvent-assisted sorption is solvation of the partner of the disrupted contact that does not directly interact with the sorbate. PMID- 11432558 TI - Bromate minimization during ozonation: mechanistic considerations. AB - Bromate formation during ozonation of bromide-containing natural waters is somewhat inversely connected to the ozone characteristics: an initial fast increase followed by a slower formation rate. During the initial phase mostly OH radical reactions contribute to bromate formation,whereas in the secondary phase both ozone and OH radicals are important. To minimize bromate formation several control options are presented: ammonia addition, pH depression, OH radical scavenging, and scavenging or reduction of hypobromous acid (HOBr) by organic compounds. Only the two first options are applicable in drinking watertreatment. By both methods a similar effect of a bromate reduction of approximately 50% can be achieved. However, bromate formation during the initial phase of the ozonation cannot be influenced by either method. Ammonia (NH3) efficiently scavenges HOBrto NH2Br. However, this reaction is reversible which leads to higher required NH3 concentrations than expected. The rate constant kNH2Br for the hydrolysis of NH2Br by OH- to NH3 and OBr- was found to be 7.5-10(6) M(-1) s(-1). pH depression shifts the HOBr/ OBr- equilibrium to HOBr and also affects the ozone chemistry. The effect on ozone chemistry was found to be more importantfor bromate formation. For a given ozone exposure, the OH radical exposure decreases with decreasing pH. Therefore, for pH depression the overall oxidation capacity for a certain ozone exposure decreases which in turn leads to a smaller bromate formation. PMID- 11432559 TI - EXAFS and XANES studies of copper in a solidified fly ash. AB - Speciation of copper in the fly ash solidification process has been studied by X ray based spectroscopies inthe present work. Fourier transformed EXAFS (extended X-ray absorption fine structural) spectra of the solidified fly ashes showed that the bond distance of Cu-O (first shell) was 1.96 A with a coordination number (CN) of about 3.0. However, in the second shell of copper atoms, the bond distance of Cu-(O)-Cu was decreased by 0.12-0.22 A during solidification, which might cause the stabilization of the CuO species in the solidified fly ash matrix. By the least-squares fits of the XANES (X-ray absorption near edge structural) spectra, fractions of the main copper species in the solidified fly ashes such as CuCl2 (0.08-0.11), Cu2O (0.07-0.09), Cu(OH)2 (0.31-0.33), and CuO (0.49-0.52) were observed. Combined EXAFS and XANES observations suggested that chemical reactions such as hydroxylation of CuCl2 and oxidation of Cu2O and/or metallic Cu might involve in the solidification process, which also led to a significant reduction of the leachability of copper from the solidified fly ashes. PMID- 11432560 TI - Sorption of pyrene to dissolved humic substances and related model polymers. 1. Structure--property correlation. AB - Solid-phase microextraction (SPME) was employed to determine sorption coefficients for pyrene on dissolved humic substances (DHS) of various origins and also on model polymers to gain a better understanding of the relationships between sorption potential and sorbent structures. The sorption potential of DHS from very different sources is described by an empirical two-parameter correlation involving the polarity (O/H atomic ratio) and the aromaticity (epsilon 280 nm) as descriptors. On the other hand, sorption experiments with well-defined model polymers, poly(acrylic acid) esters, led to the conclusion that aliphatic chains may be more effective than aromatic moieties in binding PAHs. The molecular weight of the model sorbents was found to have a significant influence on the sorption potential. PMID- 11432561 TI - Carbonyl sulfide derived from catalytic oxidation of carbon disulfide over atmospheric particles. AB - The formation of carbonyl sulfide (COS) by catalytic oxidization of carbon disulfide (CS2) over atmospheric particle catalysts was explored through FT-IR, MS (mass spectrometry), and a fixed-bed stainless steel reactor. Also the crystallizing conditions and specific surfaces (areas) of the catalysts were investigated by means of X-ray diffraction (XRD) and BET. Some oxides such as CaO, Fe2O3, Al2O3, and SiO2 were investigated under the conditions similar to the atmospheric particles as a comparison. The results showed that atmospheric particles and the oxide catalysts exhibited considerable oxidizing activity for CS2 at ambient temperature to form COS. Elemental sulfur as well as COS was one of the main products; even CO2 could be produced by a secondary reaction on some catalysts. Among the catalysts, CaO showed the strongest catalytic activity for oxidizing CS2. The catalytic activities of Fe2O3 and Al2O3 decreased considerably as compared with CaO, and SiO2 had the weakest catalytic activity. Atmospheric particles' catalytic activity is between Fe2O3's and Al2O3's. The atmospheric particles we collected mainly consist of Ca(Al2Si2O8).4H2O, which is also the main component of cement. COS, the main product, is formed by the catalytic oxidizing reaction of CS2 with adsorbed "molecular" oxygen overthe catalysts' surfaces. The concentration of adsorbed oxygen over catalysts' surfaces may be the key factor contributed to the oxidizing activities. This paper first revealed that CS2 could be catalytically oxidized over atmospheric particles to form COS. It induced that this reaction may be another important source of atmospheric COS from CS2. PMID- 11432562 TI - Determination of ozone removal rates by selected building products using the FLEC emission cell. AB - Ozone removal by 16 aged (older than 1-120 months) but unused building products or materials was studied in a test system that included the field and laboratory emission cell (FLEC). The ozone removal was studied at 50 +/- 1 ppb ozone, a relative humidity of 50 +/- 5%, a temperature of 21 +/- 2 degrees C, and an air flow rate of 900 +/- 10 mL min(-1) through the FLEC (air velocity ca. 3 cm s( 1)). The ozone removal increased rapidly during the first 1-2 min and either remained at a constant level or decreased asymptotically to reach a steady state like value. The ozone removal profiles for a given material showed good repeatability during replicate experiments. Ozone deposition velocities for the building products were calculated to be between 0.0007 cm s(-1) (lacquered ash) and 0.8 cm s(-1) (unpainted gypsum board). PMID- 11432563 TI - Estimating the long-term trend in the extreme values of tropospheric ozone using a multivariate approach. AB - This paper deals with the identification and estimation of the long-term trend in the extreme values of tropospheric ozone after allowing for the confounding effects of meteorological conditions. The U. S. Environmental Protection Agency's (EPA's) National Ambient Air Quality Standard for ozone is stated in terms of exceedances of a specified threshold level. Therefore, the EPA is concerned with the long-term trend in the probability of an exceedance. A multivariate nonparametric probit regression model estimated within the framework of a hierarchical Bayes model is used to model the probability of an exceedance after allowing for the effects of changing meteorological conditions. There are three advantages to using this model. First, the trends estimated at each site in a region can be separated into a city-wide component that is common to all sites and a site-specific component that is unique to the individual site. Second, the hierarchical Bayes framework allows for combining information across monitoring sites to increase the information available regarding the trend at each individual site. Third, the nonparametric model does not require the a priori specification of the functional forms relating the probability of an exceedance to the meteorological variables. Ozone data from four Houston, Texas monitoring sites for the period 1981-1997 are analyzed. We find that there is a downward trend in the probability of an exceedance in the 1980s followed by a relatively flat trend in the 1990s. PMID- 11432564 TI - Flow injection amperometric enzyme biosensor for direct determination of organophosphate nerve agents. AB - A flow injection amperometric biosensor for the determination of organophosphate nerve agents was developed. The biosensor incorporated an immobilized enzyme reactor that contains the enzyme organophosphorus hydrolase covalently immobilized on activated aminopropyl controlled pore glass beads and an electrochemical flow-through detector containing carbon paste working electrode, a silver/silver chloride reference electrode, and stainless steel counter electrode. The organophosphorus hydrolase catalyzed the hydrolysis of organophosphate with nitrophenyl substituent to generate p-nitrophenol which is then detected downstream electrochemically at the carbon paste electrode poised at 0.9 V vs the reference electrode. The amperometric response of the biosensor was linear up to 120 microM and 140 microM, with lower detection limits of 20 nM and 20 nM, for paraoxon and methyl parathion, respectively. The response was very reproducible (RSD 2%, n = 35) and stable for over 1 month when the immobilized enzyme column was stored at 4 degrees C. Each assay took ca. 2 min giving a sample throughput of 30 h(-1). The applicability of the biosensor to monitor paraoxon and methyl parathion in distilled water and simulated well water was demonstrated. PMID- 11432565 TI - Solid-liquid extraction laser excited time-resolved Shpol'skii spectrometry: a facile method for the direct detection of 15 priority pollutants in water samples. AB - A unique method for monitoring polycyclic aromatic hydrocarbons is reported for routine analysis of water samples. The assay consists of a three-step procedure. One hundred milliliters of water is processed through an octadecyl extraction membrane via solid-liquid extraction. The pollutants are eluted with 5 mL of n hexane and directly determined in the eluting solvent by laser excited time resolved Shpol'skii spectrometry. Seventy-seven K fluorescence measurements are made with the aid of an optical fiber probe that avoids the complications of classical low-temperature methodology. The total analysis time from the extraction to PAH identification is approximately 5 min per sample. Limits of detection are at the subparts per billion levels. The simplicity of the experimental procedure, the short analysis time, the selectivity, and the excellent analytical figures of merit demonstrate the advantages of this approach for routine analysis of water samples. PMID- 11432566 TI - Collection efficiency and capacity of three samplers for acidic and basic gases. AB - This paper compares the collection efficiency and capacity of a coated porous metal disk with those of a silica gel tube and an impinger at the acid or basic aerosol concentration of two times the permissible exposure limitation (Taiwan IOSH, Institute of Occupational Safety and Health), relative humidity of 80 +/- 5%, and temperature of 30 +/- 3 degrees C in the laboratory. Using 5% sodium carbonate/glycerin coating, the collection efficiencies of the porous metal disk for nitric acid, hydrogen chloride, and hydrogen fluoride are found to be higher than those of the silica gel tube and the impinger when the sampling time is less than 3.0 h. The gas collection capacity of the porous metal disk coated with 5% sodium carbonate is calculated to be 3.9, 5.8, and >2.5 mg for nitric acid, hydrogen chloride, and hydrogen fluoride, respectively. For ammonia, the porous metal disk coated using 4% citric acid performs slightly better than the impinger, and the corresponding capacity is slightly greater than 33.6 mg. PMID- 11432567 TI - Further developments in the use of semipermeable membrane devices (SPMDs) as passive air samplers for persistent organic pollutants: field application in a spatial survey of PCDD/Fs and PAHs. AB - Semipermeable membrane devices (SPMDs) were deployed at 19 sites in northwest England to test their efficacy as passive atmospheric samplers for polychlorinated dibenzo-p-dioxins and -furans (PCDD/Fs) and polycyclic aromatic hydrocarbons (PAHs). SPMDs were found to be efficient samplers for vapor phase species in the atmosphere, with good reproducibility between samplers. Species which are partially or completely particle associated under ambient U.K. conditions were also sampled by the SPMDs but with poorer reproducibility. It is suggested that SPMDs could be used to indicate "hotspots" of particulate associated species, however. Differences in absolute and relative concentrations of all PCDD/Fs and PAHs sequestered bythe SPMDs were observed between sites. High amounts were sequestered in SPMDs at sites where previous active monitoring has indicated relatively high atmospheric concentrations, confirming the potential of SPMDs as a tool for semiquantitative spatial monitoring of atmospheric species. SPMDs also respond to differences in the mixture of compounds present in the atmosphere, thereby aiding source apportionment studies. PMID- 11432568 TI - Identification of alkyl dimethylbenzylammonium surfactants in water samples by solid-phase extraction followed by ion trap LC/MS and LC/MS/MS. AB - A novel methodology was developed for the determination of alkyl (C12, C14, and C16) dimethylbenzylammonium chloride (benzalkonium chloride or BAC, Chemical Abstract Service number: 8001-54-5) in water samples. This method is based on solid-phase extraction (SPE) using polymeric cartridges, followed by high performance liquid chromatography/ion trap mass spectrometry (LC/MS) and tandem mass spectrometry(MS/MS) detection, equipped with an electrospray interface in positive ion mode. Chromatographic separation was achieved for three BAC homologues by using a C18 column and a gradient of acetonitrile/10 millimolar aqueous ammonium formate. Total method recoveries were higher than 71% in different water matrices. The main ions observed by LC/MS were at mass-to-charge ratios (m/z) of 304, 332, and 360, which correspond to the molecular ions of the C12, C14, and C16 alkyl BAC, respectively. The unequivocal structural identification of these compounds in water samples was performed by LC/MS/MS after isolation and subsequent fragmentation of each molecular ion. The main fragmentation observed for the three different homologues corresponded to the loss of the toluyl group in the chemical structure, which leads to the fragment ions at m/z 212, 240, and 268 and a tropylium ion, characteristic of all homologues, at m/z 91. Detection limits for the methodology developed in this work were in the low nanogram-per-liter range. Concentration levels of BAC- ranging from 1.2 to 36.6 micrograms per liter--were found in surface-water samples collected downstream from different wastewater-treatment discharges, thus indicating its input and persistence through the wastewater-treatment process. PMID- 11432569 TI - Determination of metal--EDTA complexes in soil solution and plant xylem by ion chromatography-electrospray mass spectrometry. AB - An ion chromatography-electrospray mass spectrometry (IC-MS) method was developed to quantify the metal complexes of ethylenediaminetetraacetic acid (EDTA) in soil solution and plant xylem exudate. Suitable separation of the metal-EDTA complexes was achieved on a Dionex AS5 column using 2 mM Na2CO3 as the eluant. However, satisfactory detection by eluant suppressed IC-MS, in either the positive or negative ion detection mode, could not be attained. A new eluant that still attained suitable separation and produced ionic species that could be detected by MS in the negative ion mode was developed. The eluant consisted of 2.5 mM (NH4)2CO3, 9.7 mM NH4OH, and 4% (v/v) methanol and had a pH 9.9. Even though eluant suppressed IC-MS degraded detection limits by a factor of 4 over the nonsuppressed system, using the retention time and not the m/z (mass-to-charge ratio) of the intact chelate for identification, the latter allowed the metal complexes to be detected intact and was optimized for the analysis of environmental samples. The number of metal-EDTA species that could be detected was limited by the eluant used for ion chromatography (i.e. only those complexes that were stable at high pH), with metal-EDTA complexes of Al, Cd, Cu, Co, Mn, Ni, Pb, and Zn being adequately resolved. Iron(III), Ca, MgEDTA, and EDTA itself were not detected. Detection limits for the various complexes ranged from 0.1 to 1 microM. PMID- 11432570 TI - State-of-the-art chamber facility for studying atmospheric aerosol chemistry. AB - A state-of-the-art chamber facility is described for investigation of atmospheric aerosol chemistry. Dual 28 m3 FEP Teflon film chambers are used to simulate atmospheric conditions in which aerosol formation may occur. This facility provides the flexibility to investigate dark, single oxidant reactions as well as full photochemical simulations. This paper discusses the environmental control implemented as well as the gas-phase and aerosol-phase instrumentation used to monitor atmospheric aerosol formation and growth. Physical processes occurring in the chamber and procedures for estimating secondary organic aerosol formation during reaction are described. Aerosol formation and evolution protocols at varying relative humidity conditions are presented. PMID- 11432571 TI - A new method to measure effective soil solution concentration predicts copper availability to plants. AB - Risk assessments of metal contaminated soils need to address metal bioavailability. To predict the bioavailability of metals to plants, it is necessary to understand both solution and solid phase supply processes in soils. In striving to find surrogate chemical measurements, scientists have focused either on soil solution chemistry, including free ion activities, or operationally defined fractions of metals. Here we introduce the new concept of effective concentration, CE, which includes both the soil solution concentration and an additional term, expressed as a concentration, that represents metal supplied from the solid phase. CE was measured using the technique of diffusive gradients in thin films (DGT) which, like a plant, locally lowers soil solution concentrations, inducing metal supply from the solid phase, as shown by a dynamic model of the DGT-soil system. Measurements of Cu as CE, soil solution concentration, by EDTA extraction and as free Cu2+ activity in soil solution were made on 29 different soils covering a large range of copper concentrations. Theywere compared to Cu concentrations in the plant material of Lepidium heterophyllum grown on the same soils. Plant concentrations were linearly related and highly correlated with CE but were more scattered and nonlinear with respect to free Cu2+ activity, EDTA extraction, or soil solution concentrations. These results demonstrate that the dominant supply processes in these soils are diffusion and labile metal release, which the DGT-soil system mimics. The quantity CE is shown to have promise as a quantitative measure of the bioavailable metal in soils. PMID- 11432572 TI - Adsorptive partitioning of an organic compound onto polyelectrolyte-immobilized micelles on porous glass and sand. AB - Halting the spread of organic contaminants in subsurface aquifers is a critical environmental problem. We describe a novel "permeable reactive barrier" that results when organophile-solubilizing properties are conferred on siliceous materials by treating them with a cationic polymer and oppositely charged mixed surfactant micelles. Controlled pore glass, quartz sand, and sea sand were treated with poly(diallyldimethylammonium chloride) and with mixed micelles of Triton X-100 and sodium dodecyl sulfate, either sequentially or simultaneously, following different treatment procedures. A model organophilic compound, Orange OT, was adsorbed and retained under aqueous agitation on the siliceous treated surfaces but not on untreated surfaces or those treated with micelle only. The aspect of the treatment procedure producing the most significant effect on Orange OT solubilization was the ionic strength. The retention of Orange OT in a layer of polyelectrolyte-micelle-treated sand under flow, within a column of untreated sand, demonstrates the possibility of using similar processes as a permeable reactive barrier to trap organic pollutants. PMID- 11432573 TI - Thermophilic biotrickling filtration of ethanol vapors. AB - The treatment of ethanol vapors in biotrickling filters for air pollution control was investigated. Two reactors were operated in parallel, one at ambient temperature (22 degrees C) and one at high temperature (53 degrees C). After a short adaptation phase, the removal of ethanol was similar in both reactors. At a bed contact time of 57 s, the elimination capacity exceeded 220 g m(-3) h(-1) at both temperatures. The experiments performed revealed that the process was most likely limited by biodegradation in the biofilm. The high-temperature biotrickling filter exhibited a higher degree of ethanol mineralization to CO2 (60 vs 46% at ambient temperature); hence, a lower rate of biomass accumulation was observed. Plating and cultivation of biofilm samples revealed that the high temperature biotrickling filter hosted a process culture composed of both mesophilic and thermotolerant or thermophilic microorganisms, whereas the ambient temperature reactor lacked microorganisms capable of growing at high temperature. Consequently, the performance of the control biotrickling filter was significantly affected by a short incursion at 53 degrees C. The upper temperature limit for treatment was 62 degrees C. Overall, the results of this study open new possibilities for biotrickling filtration of hot gases. PMID- 11432574 TI - Magnetic resonance angiography and evaluation of cervical arteries. AB - Multiple clinical trials have demonstrated the efficacy of endarterectomy in selected groups of patients based primarily on percent diameter stenosis. Although measurement of stenosis in the clinical trials was established by conventional angiography, there is considerable interest in noninvasive alternatives. Magnetic resonance angiography, performed using time-of-flight methods or with contrast enhancement, is one of several alternatives for noninvasive carotid evaluation. Screening examinations are routinely performed for carotid stenosis. Preoperative evaluations based on one or a combination of noninvasive tests have been proposed, although these proposals are the subject of ongoing controversy. Evaluation of the vertebral arteries is more difficult and less well studied: however, the increasing availability of therapies for posterior circulation atherosclerotic narrowing is resulting in increased interest in this problem. PMID- 11432575 TI - Limits of time-of-flight magnetic resonance angiography. AB - Many of the limits to current time-of-flight (TOF) magnetic resonance (MR) angiographic methods, including flow voids from turbulent flow, flow artifacts from pulsatile flow, and oblique flow displacement, are not inherent to the TOF enhancement mechanism and will be eliminated with technological advancements. Slowly flowing blood and physiological motion will remain an obstacle to these methods, although technological advancements will improve on current methods. Future TOF MR angiographic methods will prove to be a cost-effective competitor to MR angiographic methods requiring contrast agents in many applications. PMID- 11432576 TI - Contrast-Enhanced magnetic resonance angiography of the carotid bifurcation using the time-resolved imaging of contrast kinetics (TRICKS) technique. AB - The time-resolved contrast-enhanced magnetic resonance (MR) angiographic technique TRICKS (time-resolved imaging of contrast kinetics) reconstructs a temporal series of three-dimensional (3D) images. The temporal resolution is increased by using a short TR (<8 ms) and TE (<2 ms), zero filling, partial echo sampling, view sharing, and temporally sampling k-space at variable rates. TRICKS allows reconstruction of multiple sequential 3D volumes following bolus injection of a gadolinium chelate (0.2 mmol/kg body weight up to 40 ml, injection rate -2 ml/s). The resulting temporally defined datasets are conceptually similar to a catheter-based intra-arterial digital subtraction angiographic series, except that they are 3D volumes and not projection images. Similar to other contrast enhanced MR angiographic methods, TRICKS improves delineation of carotid artery stenosis by minimizing saturation effects. TRICKS and other contrast-enhanced MR angiographic techniques use short echo times and small voxels, thus reducing intravoxel dephasing. Surface morphology of atherosclerotic plaque and slow flow in nearly occluded vessels ("string sign") are well delineated. The major advantage of the TRICKS technique is that the timing of the acquisition in relation to the passage of the contrast bolus occurs automatically, allowing for consistent capture of the arterial phase. and eliminating the need for sophisticated synchronization methods. PMID- 11432577 TI - Magnetic resonance angiography at 3.0 Tesla: initial clinical experience. AB - Magnetic resonance (MR) angiography has undergone significant development over the past decade. It has gone from being a novelty application of MR with limited clinical use to replacing catheter angiography in some clinical applications. One of the principal limitations inherent to all MR angiographic techniques is that they remain signal limited when pushed to the limits of higher resolution and short acquisition time. Developments in magnetic gradient hardware, coil design, and pulse sequences now are well optimized for MR angiography obtained at 1.5-T main magnetic field (B-field) strength, with acquisition times and imaging matrix size near their optimal limits, respectively. Recently, the United States Food and Drug Administration (FDA) approved use of clinical magnetic resonance imaging with main magnetic field strengths of up to 4 T. Before FDA approval, use of MR with magnetic field strengths much greater than 1.5 T was essentially reserved for investigational or research applications. The main advantage of high B-field imaging is a significant improvement in the signal-to-noise ratio (SNR), which increases in an approximately linear fashion with field strength in the range of 1.5 to 3.0 T. This increased SNR is directly available when performing MR angiographic acquisitions at higher magnetic field strengths, allowing for better resolution and conspicuity of vessels with similar acquisition times. Little has been reported on the benefits of performing MR angiography at magnetic field strengths >1.5 T. The purpose of this article is to summarize our current experience with intracranial and cervical MR angiographic techniques at 3.0 T. PMID- 11432578 TI - Carotid plaque characterization by magnetic resonance imaging: review of the literature. AB - Magnetic resonance imaging (MRI) of carotid plaque has undergone significant improvements in the last decade. Early studies utilizing ex vivo specimens and spin-echo or fast spin-echo imaging led to the conclusion that T2 weighting is the best single contrast to characterize carotid plaque morphology. On these images, the fibrous plaque appears bright and the lipid core is dark; thrombus can have variable intensity. There can be an overlap in T2-weighted signal intensities among the various plaque components, which can be partially offset by the use of multispectral analysis of multiple contrast images. With improvements in coil design, sequence design, and main field and gradient capabilities, accurate in vivo differentiation and measurement of these various carotid plaque components should be possible in 3 to 5 years. Ex vivo and in vivo studies have yielded high-resolution measurements of the complex three-dimensional lumen geometry, which are being used to predict hemodynamic forces acting on the lumenal surface. Carotid plaque burden can be accurately measured in vivo today; ongoing longitudinal studies should lead to a better understanding of the relationship between plaque burden and the risk of thromboembolic complications, as well as the effect of diet and drug therapy in hyperlipidemic patients. With these developments in place or soon to be available, MRI of the diseased carotid artery wall may prove to be even more important than magnetic resonance angiography. PMID- 11432579 TI - Anaerobic biodegradation of sugar beet pulp. AB - Sugar beet pulp is a by-product of sugar production and consists mainly of cellulose, hemicellulose and pectin. Its composition is suitable for biological degradation. A possible alternative for the utilization of this material (besides cattle feeding) can be anaerobic methanogenic degradation. It has an additional advantage--biogas production. Beet pulp was treated by a two-step anaerobic process. The first step consisted of hydrolysis and acidification. The second step was methanogenesis. In this paper, observation of the process of anaerobic degradation and determination of optimal parameters is discussed. A laboratory scale model for sugar beet pulp anaerobic biodegradation was operated. Results of model performance have shown very good pulp digestion characteristics. In addition, high efficiency removal of organic matter was achieved. Methane yield was over 0.360 m3 kg(-1) dried pulp and excess sludge production was 0.094 g per gram COD added. PMID- 11432580 TI - Microbial energetics and stoichiometry for biodegradation of aromatic compounds involving oxygenation reactions. AB - Oxygenation reactions significantly alter the energy and electron flows and, consequently, the overall stoichiometry for the microbial utilization of aromatic compounds. Oxygenation reactions do not yield a net release of electrons, but require an input of electrons to reduce oxygen molecules. The biodegradation pathway of phenanthrene as a model compound was analyzed to determine the impact of oxygenation reactions on overall stoichiometry using the half-reaction method. For individual oxygenation reactions, the half-reaction method for analyzing the electron and energy flows must be modified, because the reactions do not release electrons for synthesis or energy generation. Coupling the oxygenation reaction to subsequent reaction steps provides a net electron release for the coupled reactions. Modeling results indicate that oxygenation reactions increase the oxygen requirement and reduce the cell yield, compared to the conventional mineralization represented by hydroxylation reactions in place of oxygenations. The computed yields considering oxygenation reactions conform better to empirical yields reported in the literature than do yields computed by the hydroxylation single-step methods. The coupled-reaction model also is consistent with information about the ways in which micro-organisms that degrade aromatics accumulate intermediates, regulate degradation genes, and organize enzyme clusters. PMID- 11432581 TI - Hydrolysis of newspaper polysaccharides under sulfate reducing and methane producing conditions. AB - The initial decomposition rates of cellulose and hemicellulose were measured using toluene to specifically inhibit the microbial uptake of hydrolysis products during the degradation of newspaper under sulfate reducing and methane producing conditions. The amount of glucose and xylose accumulation in the first 2 weeks of incubation period was higher in the sulfate reducing condition compared to the methane producing condition. It was estimated that 28 and 6% of initially loaded cellulose in the sulfate reducing condition and the methane producing condition was hydrolyzed, respectively. Accordingly, the newspaper-cellulose hydrolysis rate constant was estimated to be 6.7 times higher in sulfate reducing condition than in methane producing condition. Based on the glucose accumulation patterns, when sulfate reducing bacteria (SRB) were inhibited by anthraquinone and molybdate (Na2MoO4), it may be suggested that SRB might have contributed to the hydrolysis of cellulose, while their effect on the hydrolysis of hemicellulose could not be elucidated. PMID- 11432582 TI - Mineralization of 1,4-dioxane in the presence of a structural analog. AB - A mixed culture with the ability to aerobically biodegrade 1,4-dioxane in the presence of tetrahydrofuran (THF) was enriched from a 1,4-dioxane contaminated aquifer. This consortium contained 3-4 morphologically different types of colonies and was grown in mineral salts media. Biodegradation of 1,4-dioxane began when THF concentrations in batch experiments became relatively low. No biodegradation of 1,4-dioxane was observed in the absence of THF and the measured cell yield was similar during degradation of 1,4-dioxane with THF or with THF alone. However, when the consortium was grown in the presence of 14C-1,4-dioxane plus THF, 2.1% of the radiolabeled 1,4-dioxane was present in the particulate fraction. The majority of the 14C (78.1%) was recovered as 14CO2, while 5.8% remained in the liquid fraction. This activity is interesting since the non growth substrate is mineralized, yet only minimally assimilated into biomass. Using THF as the growth substrate, the consortium also degraded 1,3-dioxane, methyl t-butyl ether, ethyl t-butyl ether and t-amyl methyl ether. PMID- 11432583 TI - Pathways for 3-chloro- and 4-chlorobenzoate degradation in Pseudomonas aeruginosa 3mT. AB - A bacterial isolate, Pseudomonas aeruginosa 3mT exhibited the ability to degrade high concentrations of 3-chlorobenzoate (3-CBA, 8 g l(-1)) and 4-chlorobenzoate (4-CBA 12 g l(-1)) (Ajithkumar 1998). In this study, by delineating the initial biochemical steps involved in the degradation of these compounds, we investigated how this strain can do so well. Resting cells, permeabilised cells as well as cell-free extracts failed to dechlorinate both 3-CBA and 4-CBA under anaerobic conditions, whereas the former two readily degraded both compounds under aerobic conditions. Accumulation of any intermediary metabolite was not observed during growth as well as reaction with resting cells under highly aerated conditions. However, on modification of reaction conditions, 3-chlorocatechol (3-CC) and 4 chlorocatechol (4-CC) accumulated in 3-CBA and 4-CBA flasks, respectively. Fairly high titres of pyrocatechase II (chlorocatechol 1,2-dioxygenase) activity were obtained in extracts of cells grown on 3-CBA and 4-CBA. Meta-pyrocatechase (catechol 2,3-dioxygenase) activity against 4-CC and catechol, but not against 3 CC, was also detected in low titres. Accumulation of small amounts of 2-chloro-5 hydroxy muconic semialdehyde, the meta-cleavage product of 4-CC, was detected in the medium, when 4-CBA concentration was 4 mM or greater, indicating the presence of a minor meta-pathway in strain 3mT. However, 3-CBA exclusively, and more than 99% of 4-CBA were degraded through the formation of the respective chlorocatechol, via a modified ortho-pathway. This defies the traditional view that the microbes that follow chlorocatechol pathways are not very good degraders of chlorobenzoates. 4-Hydroxybenzoate was readily (and 3-hydroxybenzoate to a lesser extent) degraded by the strain, through the formation of protocatechuate and gentisate, respectively, as intermediary dihydroxy metabolites. PMID- 11432584 TI - Effects of surfactant addition on the biomineralization and microbial toxicity of phenanthrene. AB - Surfactants are known to increase the apparent aqueous solubility of polycyclic aromatic hydrocarbons and may thereby enhance their bioavailability. In this study the effects of four surfactants on the mineralization of phenanthrene by Pseudomonas aeruginosa in liquid culture and in soil-water suspensions was studied in batch reactors over a 15-week study period. In the absence of surfactant, liquid cultures mineralized approximately 50% of the phenanthrene added within seven weeks following a one-week lag period and an initial mineralization rate of 0.04 mg/d. Mineralization in soil-water suspensions proceeded without any measurable lag period. The initial mineralization rate was lower (0.006 mg/d), but mineralization continued to >70% over the fifteen week period. In general, the addition of very low concentrations of surfactant (<0.001%) to liquid cultures did not impact mineralization significantly. At higher surfactant concentrations (>CMC) all surfactants were seen to be inhibitory. In soil-water systems, the rate of phenanthrene mineralization was decreased even at surfactant doses that did not produce significant solubilization. In summary, none of the surfactants enhanced the mineralization of phenanthrene by P. aeruginosa in liquid culture or in soil-water suspensions. In order to rank surfactant toxicity, microbial toxicity tests were performed measuring the light output of bioluminescent bacteria as affected by the presence of surfactants. Additional toxicity testing indicated that the presence of solubilized phenanthrene increased the toxicity of the surfactant by a 100-fold suggesting that the toxicity of solubilized substrate needs also to be considered in the application of surfactant-amended remediation. PMID- 11432585 TI - Interleukin-10 inhibits polymethylmethacrylate particle induced interleukin-6 and tumor necrosis factor-alpha release by human monocyte/macrophages in vitro. AB - Periprosthetic membranes commonly observed at sites of total joint implant loosening exhibit abundant macrophages and particulate debris. Macrophages phagocytose orthopedic debris and release the pro-inflammatory mediators interleukin-1, interleukin-6, tumor necrosis factor-alpha, and prostaglandin E2. Populations of activated lymphocytes are often seen in periprosthetic membranes. These lymphocytes may modulate the monocyte/macrophage response to particulate debris and influence aseptic loosening. In addition, other immunologic agents, such as interleukin-10, are present in tissues harvested from the bone-implant interface of failed total joint arthroplasties. The present study examined the effects of interleukin-10 on polymethylmethacrylate (PMMA) particle challenged human monocyte/macrophages in vitro. Human monocyte/macrophages isolated from buffy coats of five healthy individuals were exposed to 1-10 microm PMMA particles. Interleukin-10 was added to the monocyte/macrophages with and without the addition of PMMA particles. Interleukin-10-induced alterations in monocyte/macrophage metabolism were determined measuring interleukin-6 and tumor necrosis factor-alpha release by the cells following exposure to PMMA particles. Exposure of the monocyte/macrophages to PMMA particles resulted in a dose dependent release of interleukin-6 and tumor necrosis factor-alpha at 48 h. Interleukin-10 reduced the levels of interleukin-6 and tumor necrosis factor alpha release by macrophages in response to PMMA particles in a dose-dependent manner. At 48 h, particle-induced interleukin-6 release was inhibited by 60 and 90% with 1.0 and 10.0 ng/ml treatments of interleukin-10, respectively. At 48 h, particle-induced tumor necrosis factor-alpha release was inhibited by 58 and 88% with 1.0 and 10.0 ng/ml treatments of interleukin-10, respectively. Interleukin 10 challenge alone did not significantly alter basal interleukin-6 or tumor necrosis factor-alpha release relative to control cultures. The data presented in this study demonstrate that the anti-inflammatory cytokine, interleukin-10, inhibits monocyte/macrophage release of the pro-inflammatory cytokines interleukin-6 and tumor necrosis factor-alpha in response to PMMA particle challenge in vitro. PMID- 11432586 TI - In vitro gene delivery mediated by chitosan. effect of pH, serum, and molecular mass of chitosan on the transfection efficiency. AB - Aminopolysaccharides such as chitosan and polygalactosamine (pGalN) were used to transfer luciferase plasmid into tumor cells. Chitosan largely enhanced the transfection efficiency of luciferase plasmid (pGL3), while pGalN did not at all. Transfection efficiencies of the pGL3/chitosan complexes were dependent on pH of culture medium, stoicheometry of pGL3:chitosan, serum, and molecular mass of chitosan. Transfection efficiency at pH 6.9 was higher than that at pH7.6. Optimum charge ratio of the pGL3:chitosan was 1:5. Chitosan polymer of 15 and 52 kDa largely promoted luciferase activities. Transfection efficiency mediated by chitosan of > 100 kDa was less than that by chitosan of 15 and 52 kDa. Heptamer (1.3 kDa) did not show any gene expression. Cationic liposome (lipofectin) associated gene expression was inhibited by serum, while chitosan showed resistance to serum. PMID- 11432587 TI - Release of recombinant human bone morphogenetic protein-2 from heterocyclic methacrylate polymer systems. AB - The release of recombinant human bone morphogenetic protein-2 (rhBMP-2) from three room temperature polymerising methacrylate systems has been studied. These all contained poly(ethyl methacrylate) powder, but the monomer liquids comprised, respectively, tetrahydrofurfuryl methacrylate (THFM), 90/10 THFM/hydroxyethyl methacrylate (HEMA), and 70/30 THFM/ HEMA. In all cases, rhBMP-2 was released, but the addition of 10% HEMA accelerated release (a nine-fold increase in diffusion coefficient); a further increase to 30% HEMA had no additional effect. For most of the release process, a diffusion process operated, although the early stages were not well defined. At the end of the 15 day period, the release, respectively, for the PEM/THFM, PEM:90/10 THFM/HEMA and PEM:70/30 THFM/HEMA systems was 596, 878 and 923 ng (i.e. up to 92% of the rhBMP-2 added). PMID- 11432588 TI - Blends of isoprene-styrene/methacrylate monomer systems as denture soft lining material. AB - This work further develops the concept of using an elastomer gelled with methacrylate monomers to produce a methacrylate-based soft lining material without the use of a plasticizer. An isoprene-styrene (SIS) block copolymer was mixed with methyl methacrylate (MMA) and 1,6-hexandiol dimethacrylate (HDMA). The HDMA was used as a cross-linking agent. The elastomer/monomer ratios were maintained at 50/50 whereas the monomers ranged from 0 to 100%) HDMA. Mechanical properties and water absorption/desorption characteristics were used to assess the effect of varying the monomer compositions. The results indicated that phase separation took place, in particular at high HDMA content. This significantly increased the Young's modulus and decreased the elongation to break. Generally, the water uptake tended to decrease with increasing HDMA content, reflecting the effect of modulus. Second absorption cycles gave higher uptake values compared to the first. Formulations with a high amount of HDMA gave materials with modulus values too high for soft lining applications. This suggests that the optimum formulation requires a compromise between modulus and water uptake. PMID- 11432589 TI - Basic fibroblast growth factor enhances in vitro mineralization of rat bone marrow stromal cells grown on non-woven hyaluronic acid based polymer scaffold. AB - A biodegradable non-woven hyaluronic acid polymer scaffold (Hyaff 11) was analysed in vitro as a carrier vehicle for differentiation and mineralization of rat bone marrow stromal cells (BMSC). BMSC were grown on Hyaff 11 in a mineralizing medium in the presence/absence of basic fibroblast growth factor (bFGF). Osteoblastic differentiation was investigated by light and electron microscopy analysing the expression of osteogenic markers: calcium, alkaline phosphatase (AP), osteopontin (OP), bone sialoprotein (BSP) and collagen type 1. We also measured proliferation, AP activity and mRNA expression of AP and osteocalcin (OC). Electron microscopy and Toluidine-blue staining demonstrated that bFGF accelerated (day 20 vs. day 40) and increased mineralization. With bFGF, calcium, OP and BSP were strongly enhanced at day 40, whereas AP decreased. Our in vitro results demonstrate that Hyaff 11 is a useful vehicle for growth, differentiation and mineralization of rat BMSC, and that it permits bone development. PMID- 11432590 TI - Fracture behavior of UHMWPE in non-implanted, shelf-aged knee prostheses after gamma irradiation in air. AB - We studied non-implanted ultra-high molecular weight polyethylene (UHMWPE) knee components, that were gamma-sterilized in air and shelf-aged during 6 and 7 years, to explore fracture mechanisms in the degraded material. Prior to the fractography study, we assessed degradation through FTIR and optical density measurements in microtome sections. Then, we morphologically compared the surface fractography of degraded, oxidized UHMWPE with that of non-degraded material in specimens from the same implants, fractured after uniaxial tensile tests in standardized samples. Fractography surface analysis clearly differentiated the behavior of non-degraded and degraded UHMWPE. Failure in non-degraded samples occurred through craze formation, followed by fast crack propagation. Quantitative fracture analysis suggested a ductile behavior. This was supported by the identified local plasticity in the region close to the initial flaw. In the fractography surface analysis of degraded material, a fibrillar behavior was observed. A heterogeneous domain structure appeared, showing different coarseness with multiple void sizes. Our results, based on mechanical measurements and fracture morphology in the subsurface region, confirmed the embrittlement of oxidized UHMWPE. PMID- 11432591 TI - Immobilization of poly(ethylene glycol) or its sulfonate onto polymer surfaces by ozone oxidation. AB - A novel surface modification method has been developed to improve biocompatibility of polymeric biomaterials. This approach involves ozonation and then followed by graft polymerization with acrylates containing PEG, sulfonated PEG or by coupling of PEG derivatives. All the reactions were confirmed by ATR FT IR and ESCA. The degree of ozonation measured by the iodide method was dependent on the ozone permeability of the polymers used. Surface hydrophilicity was investigated by measuring the contact angles. Ozonation itself yielded a slight increase in hydrophilicity and a decrease in platelet adhesion, but PEG immobilization showed a significant effect on surface hydrophilicity and platelet adhesion to confirm well-known PEG's passivity which minimize the adhesion of blood components on polymer surfaces. Both graft polymerization and coupling were effective for PU. In contrast, only grafting gave enough yields for PMMA and silicone. Platelet adhesion results demonstrated that all PEG modified surfaces adsorbed lower platelet adhesion than untreated or ozonated ones. Polymers coupled with sulfonated PEG exhibited the lowest platelet adhesion when compared with control and PEG coupled ones by virtue of the synergistic effect of non adhesive PEG and negatively charged SO3 groups. This PEG or sulfonated PEG immobilization technology using ozonation is relatively simple for introducing uniform surface modification and therefore very useful for practical application of blood contacting medical devices. PMID- 11432592 TI - Evaluation effects of chitosan for the extracellular matrix production by fibroblasts and the growth factors production by macrophages. AB - Chitosan is reported as an accelerator of wound healing. Histological findings of previous reports indicate that chitosan accelerates the reformation of connective tissue, however the details of the mechanism are not clear. In this study, firstly L929 mouse fibroblasts were cultured with chitosan and the production of extracellular matrix (ECM) was evaluated in vitro. Type I and III collagens and fibronectin were secreted by L929 with or without chitosan; however there was no significant difference in the amount of ECM between the control and the chitosan groups. Secondly, macrophages were stimulated with chitosan, and then transforming growth factor-beta 1 (TGF-beta1) and platelet-derived growth factor (PDGF) messenger ribonucleic acid (mRNA) expressions and production of their proteins were assayed in vitro. As a result, chitosan promoted the production of TGF-beta1 and PDGF. These results indicate that chitosan does not directly accelerate ECM production by fibroblast and the ECM production may increase by the growth factors. PMID- 11432593 TI - Structure and surface of TiNi human implants. AB - The surface and the "bulk" structure of TiNi implants were characterized using scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray photoemission spectroscopy (XPS), and scanning Auger microprobe analysis (AES). TiNi implants were compared with otherwise identically prepared non-implanted specimens, and sputter-cleaned and reoxidized samples. Non-implanted and implanted samples had essentially the same surface topography and microstructure. Ti, O, and C were the dominant elements detected on the surface. Trace amounts (approximately 1 at%) of Ni and Ca, N, Si, B, and S were also detected. Ti was present as TiO2 on the surface, while nickel was present in metallic form. A significant difference in Ni peak intensity was observed when retrieved or non implanted control samples (a very low nickel content) were compared with sputter cleaned and reoxidized samples (well-detected nickel). It is evident that the method of passivation is crucial for nickel loosening. No major changes occurred in the TiNi samples bulk structure or in the surface oxide during the implantation periods investigated. PMID- 11432594 TI - Effect of calcium-ion implantation on the corrosion resistance and biocompatibility of titanium. AB - This work presents data on the structure and corrosion resistance of titanium after calcium-ion implantation with a dose of 10(17) Ca+/cm2. The ion energy was 25 keV. Transmission electron microscopy was used to investigate the microstructure of the implanted layer. The chemical composition of the surface layer was examined by XPS and SIMS. The corrosion resistance was examined by electrochemical methods in a simulated body fluid (SBF) at a temperature of 37 degrees C. Biocompatibility tests in vitro were performed in a culture of human derived bone cells (HDBC) in direct contact with the materials tested. Both, the viability of the cells determined by an XTT assay and activity of the cells evaluated by alkaline phosphatase activity measurements in contact with implanted and non-implanted titanium samples were detected. The morphology of the cells spread on the surface of the materials examined was also observed. The results confirmed the biocompatibility of both calcium-ion-implanted and non-implanted titanium under the conditions of the experiment. As shown by TEM results, the surface layer formed during calcium-ion implantation was amorphous. The results of electrochemical examinations indicate that calcium-ion implantation increases the corrosion resistance, but only under stationary conditions; during anodic polarization the calcium-ion-implanted samples undergo pitting corrosion. The breakdown potential is high (2.7-3 V). PMID- 11432595 TI - On the transformation behaviour, mechanical properties and biocompatibility of two niti-based shape memory alloys: NiTi42 and NiTi42Cu7. AB - The transformation behaviour, mechanical properties and cytotoxicity of a binary NiTi42 and a ternary NiTi42Cu7 alloy have been investigated. The transformation temperatures were determined via differential scanning calorimetry, the mechanical properties have been investigated in 3-point bending tests in the temperature range between 6 and 60 degrees C. The cytotoxicity tests were performed on both alloys in cultured epithelial cells from human gingiva. The cytotoxicity investigations included both MTT tests and morphological observations. It is shown that although the ternary alloy is characterised by a narrower hysteresis and superior mechanical properties, including fatigue resistance, its cytotoxicity is higher than that of the binary alloy. This is thought to arise from the release of copper ions in the medium, which upon atomic absorption spectroscopy measurements amount to approximately 2.8 microg cm(-2) for an incubation period of 7 days. PMID- 11432596 TI - In vitro release of heparin from silica xerogels. AB - Heparin, a powerful anticoagulant used for the prophylaxis of both surgical and medical thrombosis, was incorporated into a silica xerogel matrix during polycondensation of organic silicate. The influence of various chemical sol-gel parameters (the properties of reaction precursors, catalyst and final moisture content of the gel and heparin concentration) was studied. The release of heparin from the gel was according to zero order during the dissolution period and the release rate of heparin was proportional to the drug load in the concentration range between 6.8 and 13.6 wt%. It was found that the catalyst used for the preparation of the gel, the final moisture content and the chemical modification of silica xerogel network have an influence on the release rate of heparin. The released heparin from all the different xerogels studied retained about 90% of its biological activity. PMID- 11432597 TI - Influence of thickness on the properties of hydroxyapatite coatings deposited by KrF laser ablation. AB - The growth of hydroxyapatite coatings obtained by KrF excimer laser ablation and their adhesion to a titanium alloy substrate were studied by producing coatings with thicknesses ranging from 170 nm up to 1.5 microm, as a result of different deposition times. The morphology of the coatings consists of grain-like particles and also droplets. During growth the grain-like particles grow in size, partially masking the droplets, and a columnar structure is developed. The thinnest film is mainly composed of amorphous calcium phosphate. The coating 350nm thick already contains hydroxyapatite, whereas thicker coatings present some alpha tricalcium phosphate in addition to hydroxyapatite. The resulting coating to substrate adhesion was evaluated through the scratch test technique. Coatings fail under the scratch test by spallating laterally from the diamond tip and the failure load increases as thickness decreases, until not adhesive but cohesive failure for the thinnest coating is observed. PMID- 11432598 TI - The effect of cross-linking agents on acrylic bone cements containing radiopacifiers. AB - Poly(methylmethacrylate) PMMA, based cements are the most widely used bone cements in joint replacement surgery. Although, there are some disadvantages in the use of these cements, the clinical success rate is fairly high. Intrinsic radiopacity is difficult to achieve in these cements due to the constituent elements of the PMMA polymer. As radiopacity is an essential requirement, PMMA bone cements have been rendered radiopaque by blending heavy metal ion salts, which tend to adversely affect the mechanical and biological properties of the bone cement. In this study, dimethacrylate cross-linking agents were added to the monomer phase in order to generate a cross-linked matrix, with barium sulphate as a radiopaque agent. The results suggest that the mechanical properties can be improved or retained with the addition of such cross-linking agents. PMID- 11432599 TI - The impact of adolescent girls' life concerns and leisure activities on body dissatisfaction, disordered eating, and self-esteem. AB - The aim of this study was to situate adolescent girls' body dissatisfaction, disordered eating, and self-esteem in the context of their life concerns and leisure activities. Questionnaires containing measures of life concerns, leisure activities, body dissatisfaction, disordered eating, and self-esteem were administered to 306 girls with a mean age of 16 years. It was found that although academic success and intelligence were rated as the most important life concerns, an emphasis on slimness was most strongly linked to body dissatisfaction, disordered eating, and global self-esteem. An emphasis on popularity with girls also was related to body dissatisfaction, and hours spent watching television were related to lower self-esteem. In contrast, emphasis on sport seemed to serve a protective function. It was concluded that adolescent girls who have a high concern for slimness should be assisted in decreasing this emphasis in order to improve their general well-being. PMID- 11432600 TI - Motor development during the first year: a comparative study. AB - This investigation of the motor development characteristics of Brazilian infants during the 1st year of postnatal life compared the findings with those of a widely used U.S. sample (N. Bayley, 1993). Analysis of the 12 monthly assessments indicated that Brazilian infants' scores increased with age, with the greatest increase occurring over the first 8 months. As a general comparison, the results are similar to the U.S. patterns of development. However, the Brazilian sample exhibited mean scores significantly lower than the U.S. sample's in the 3rd, 4th, and 5th months. More specific analyses indicated that the difference within these months was on tasks of sitting and grasping. Possible explanations for the differences focus on variations in child-rearing practices and the influence of biological maturation in early movement behaviors. PMID- 11432601 TI - Action perception and outcome valence: effects on children's inferences of intentionality and moral and liking judgments. AB - The influences of different action-outcome scenarios on children's evaluative judgments and inferences of outcome intentionality were assessed. One hundred forty-five kindergartners, 2nd graders, and 4th graders heard 4 stories about child actors who engaged in 1 action or 3 equifinal actions and caused a positive or negative outcome. The stories made no mention of the actors' anticipated outcome so that we could assess the children's inferences of whether the actors wanted and had tried to cause the outcome. Children also rated their liking for the actors and the actors' morality. Children's moral and liking judgments were not significantly differentiated by action condition. However, actors who caused positive outcomes received favorable liking and moral judgments, and actors who caused negative outcomes received neutral liking and moral judgments. Children's intentionality inferences varied by the actors' actions and were moderated by outcome valence. The authors discuss children's apparent use of the valence rule when inferring intentionality and their reluctance to judge harshly actors who cause negative outcomes when not privy to the actors' intentions. PMID- 11432602 TI - Peer rejection in childhood: effects on maternal depression and behavior problems in toddlers. AB - The aim of this study was to develop a conceptual model that would clarify how a woman's recollections of peer and parent rejection in childhood influence her emotional well-being and, resultantly, her own child's behavior problems. Given current interest in peer versus parent influences, the authors used a design and analysis to reveal the unique contributions of peer and parent rejection. The participants were a community sample of 88 mothers and their 2 1/2-year-old children. The study found that mothers' recollections of peer rejection in childhood were significantly associated with depressive symptomatology, but recollections of parental rejection in childhood were unrelated to current depressive symptoms. Recollections of peer rejection during childhood, internal representation of self, and depressive symptoms made significant, unique contributions to the variance in behavior problems in their children. Together, these variables significantly explained 27% of the variance in the final model. PMID- 11432603 TI - The leftward cradling bias and prosody: an investigation of cradling preferences in the deaf community. AB - Popular theory on the tendency to cradle an infant to the left side points to the specialization of the right hemisphere for the perception and expression of emotion. J. S. Sieratzki and B. Woll (1996) recently suggested that more emphasis be placed on the auditory modality, specifically focusing on the role of prosodic information. In this study, the direction of the lateral cradling bias in a group of profoundly deaf children, a group of deaf adults, and a control group of adults with no hearing impairment was investigated. The authors found a strong leftward cradling bias in all groups, a bias that was, if anything, stronger in the deaf participants. Given that people who are profoundly deaf, especially those who have been deaf from birth, have not been exposed to auditory prosody, the data do not support the suggestion that such prosodic information is the basis for the leftward cradling bias. PMID- 11432604 TI - Demand characteristics and self-report measures of imaginary audience sensitivity: implications for interpreting age differences in adolescent egocentrism. AB - Self-consciousness during early adolescence has been explained as an outcome of adolescent egocentrism, in which adolescents create an imaginary audience (IA) of attentive, critical peers. The possibility that such self-consciousness might result from contact with peers who are more attentive and critical than those encountered during childhood or adulthood has not been considered. Study 1 tested whether young adults, who are not theoretically susceptible to IA, could be made to receive high scores on IA and self-consciousness measures by having them complete a procedure in 1 of 3 laboratory conditions-a critical audience, a noncritical audience, or no audience. However, participants in the critical audience condition received significantly lower IA and self-consciousness scores than participants in the no-audience condition did. Study 2 tested whether the directions given to Study 1 participants might have been responsible for the unexpected findings. Results indicated that participants instructed to give mature-sounding responses received lower IA/self-consciousness scores than did those asked to report their honest opinions. Together, the results of Studies 1 and 2 indicated that survey measures of IA are subject to demand characteristics and highlighted the need to interpret with caution age differences in IA as traditionally assessed. PMID- 11432605 TI - Recognition of emotion in facial expressions and vocal tones in children with psychopathic tendencies. AB - The authors investigated the ability of children with emotional and behavioral difficulties, divided according to their Psychopathy Screening Device scores (P. J. Frick & R. D. Hare, in press), to recognize emotional facial expressions and vocal tones. The Psychopathy Screening Device indexes a behavioral syndrome with two dimensions: affective disturbance and impulsive and conduct problems. Nine children with psychopathic tendencies and 9 comparison children were presented with 2 facial expression and 2 vocal tone subtests from the Diagnostic Analysis of Nonverbal Accuracy (S. Nowicki & M. P. Duke, 1994). These subtests measure the ability to name sad, fearful, happy, and angry facial expressions and vocal affects. The children with psychopathic tendencies showed selective impairments in the recognition of both sad and fearful facial expressions and sad vocal tone. In contrast, the two groups did not differ in their recognition of happy or angry facial expressions or fearful, happy, and angry vocal tones. The results are interpreted with reference to the suggestion that the development of psychopathic tendencies may reflect early amygdala dysfunction (R. J. R. Blair, J. S. Morris, C. D. Frith, D. I. Perrett, & R. Dolan, 1999). PMID- 11432606 TI - Children's perceptions of unfair reward and punishment. AB - Being able to judge the fairness of a personal encounter and having an appreciation of the associated feelings are important components of prosocial development. This study explored a common feature of everyday experience: unfair reward and unfair punishment. Scenarios depicting 4 possible variants of unfairness were read to children aged 9 to 11 years, who then made judgments regarding the degree of unfairness and the nature and strength of the feelings experienced by the characters. Our hypothesis that children with classroom conduct problems would judge the non-receipt of a deserved reward as worse than the receipt of an undeserved punishment was not confirmed. This differentiation, however, did prove to be characteristic of boys in general, but not girls. Being asked to think of unfair things that had actually happened did not appear to influence the children's responses to hypothetical unfair situations, but did reveal that children experience and remember a variety of unfair events in everyday family contexts. This study provides evidence that children actively monitor the receipt of social reward and punishment according to their perception of fairness. PMID- 11432607 TI - Aggression in toddlers: associations with parenting and marital relations. AB - This study examined the relation among parenting factors, marital relations, and toddler aggression. A structured questionnaire was administered to both parents of 254 2-year-olds. The authors used correlation and hierarchical multiple regression analyses to assess the extent to which certain personality traits, drug use, parenting style, and marital conflicts were related to the toddlers' aggressive behavior. Results showed that the maternal child-rearing and parental aggression domains had a direct effect on toddler aggression. The domain of maternal child rearing also served as a mediator for the domains of marital relations, paternal child rearing, parental aggression, and parental drug use. The findings indicated that maternal child-rearing practices, personality attributes, and drug use were more important than paternal attributes in relation to toddler aggression. Implications for prevention among families at risk are discussed. PMID- 11432608 TI - The profile variability index in relation to psychopathology and academic achievement. PMID- 11432609 TI - Cheap drugs? Part 3: Bush continues Clinton policy on AIDS drugs but... PMID- 11432610 TI - Cheap drugs? Part 4: Blair sides with industry or simply protects intellectual property? PMID- 11432611 TI - The virtual supercomputer: science-fiction becomes a reality against cancer. PMID- 11432612 TI - High-dose chemotherapy for ovarian cancer: are we ready to go? PMID- 11432613 TI - The role of cisplatin in the treatment of small-cell lung cancer? PMID- 11432614 TI - Risk assessment in ALL in children: a focus on PCR-based techniques for MRD detection. PMID- 11432615 TI - Paclitaxel: epirubicin in metastatic breast cancer--a review. AB - BACKGROUND: Treatment of metastatic breast cancer (MBC) with paclitaxel (T) and doxorubicin has yielded high response rates but the regimen is associated with significant cardiac toxicity. Epirubicin (E) is a less cardiotoxic anthracycline which has also been combined with paclitaxel in the treatment of MBC. MATERIALS AND METHODS: This paper is a review of studies evaluating the pharmacokinetics, toxicity profile, and efficacy of the ET combination in MBC. RESULTS: The ET combination has been studied extensively in Europe. The unique pharmacokinetics of the combination do not lead to the accumulation of cardiotoxic metabolites as in the case of the doxorubicin-paclitaxel combination. In terms of efficacy, the ET combination yields an overall response rate of 50%-70% and complete response rate (CR) 10%-15% in MBC in the same range as the more recent doxorubicin paclitaxel studies. CONCLUSION: In summary the ET combination is safe and effective in MBC. It is less cardiotoxic than the doxorubicin paclitaxel combination. Further studies with ET in both the adjuvant setting and in MBC are in progress. PMID- 11432616 TI - Phase I and pharmacokinetic study of hepatic arterial infusion with oxaliplatin in combination with folinic acid and 5-fluorouracil in patients with hepatic metastases from colorectal cancer. AB - BACKGROUND: To determine dose-limiting toxicity (DLT), maximum tolerated dose (MTD), and pharmacokinetics (PK) of oxaliplatin administered as hepatic arterial infusion. PATIENTS AND METHODS: Patients with isolated hepatic metastases from colorectal cancer were treated every three weeks with increasing doses of oxaliplatin (4 hours; starting dose 25 mg/m2, escalation in steps of 25 mg/m2) in combination with folinic acid (1 hour, 200 mg/m2) and 5-fluorouracil (2 hour, 600 mg/m2). RESULTS: Twenty-one patients (median age, 61 years) have been entered all of whom are fully evaluable. The DLT has been observed at dose level 6, i.e., at 150 mg/m2/cycle and consisted of leucopenia, obliteration of the hepatic artery, and acute pancreatitis. Overall, toxicity mainly consisted of nausea/vomiting (16 of 21 patients), anemia (16 of 21), upper abdominal pain (15 of 21), sensory neuropathy (10 of 21), diarrhea (9 of 21), and thrombocytopenia (9 of 21). The mean PK parameters were: terminal half-life of ultrafiltrable platin, 17.75 +/- 9.29 hours; renal elimination, 48.7% +/- 14.1% of the applied dose; renal clearance 135.55 +/- 45.32 ml/min. The mean area under the plasma-concentration curve (AUC) increased linearly from 3.22 +/- 0.61 microg x h/ml to 18.45 +/- 8.90 microg x h/ml through the first five dose levels (P = 0.0004). Ten of eighteen evaluable patients achieved a complete or partial response (59%). CONCLUSIONS: The recommended dose for phase II studies is 125 mg/m2 oxaliplatin. PMID- 11432617 TI - A phase I and pharmacologic study of capecitabine and paclitaxel in breast cancer patients. AB - BACKGROUND: Based on preclinical studies demonstrating that treatment with paclitaxel upregulates intratumoral thymidine phosphorylase (dTHdPase), which catalyzes the final step in the conversion of the oral fluoropyrimidine capecitabine to 5-fluorouracil (5-FU), as well as the overlapping spectra of activity for these agents, particularly in metastatic breast cancer, this phase I study evaluated the feasibility of administering capecitabine on an intermittent schedule in combination with paclitaxel in previously-treated patients with locally advanced or metastatic breast cancer. The study also sought to recommend doses for subsequent disease-specific studies, identify clinically significant pharmacokinetic interactions, and detect preliminary antitumor activity. PATIENTS AND METHODS: Nineteen previously treated women with metastatic breast cancer whose prior treatment included neither paclitaxel or capecitabine received one hundred one courses of capecitabine and paclitaxel. Paclitaxel was administered as a three-hour intravenous (i.v.) infusion at a fixed dose of 175 mg/m2 and capecitabine was administered as 2 divided daily doses for 14 days followed by a seven-day rest period every 3 weeks. The dose of capecitabine was increased from a starting dose of 1650 mg/m2/d. The plasma sampling scheme in the first course permitted characterization of the pharmacokinetics of each agent given alone and concurrently to detect major pharmacokinetic interactions. RESULTS: Palmar plantar erythrodysesthesia (hand foot syndrome) and neutropenia were the principal dose-limiting toxicities (DLT). Other toxicities included diarrhea and transient hyperbilirubinemia. Three of eight new patients treated with capecitabine 2000 mg/m2/d and paclitaxel 175 mg/m2 experienced DLT in the first course, whereas none of eleven new patients treated with capecitabine 1650 mg/m2/d and paclitaxel 175 mg/m2 developed DLT. Pharmacokinetic studies indicated that capecitabine did not grossly affect the pharmacokinetics of paclitaxel, and there were no major effects of paclitaxel on the pharmacokinetics of capecitabine and capecitabine metabolites. However, AUC values for the major 5-FU catabolite, fluorobeta-alanine (FBAL), were significantly lower in the presence of paclitaxel. Two complete and seven partial responses (56% response rate) were observed in sixteen patients with measurable disease; four of six patients whose disease was previously treated with high-dose chemotherapy and hematopoietic stem cell support had major responses. Seven of nineteen patients had stable disease as their best response. CONCLUSIONS: Recommended combination doses of capecitabine on an intermittent schedule and paclitaxel are capecitabine 1650 mg/m2/d orally for 14 days and paclitaxel 175 mg/m2 i.v. every 3 weeks. The favorable preclinical interactions between capecitabine and paclitaxel, as well as the acceptable toxicity profile and antitumor activity in patients with metastatic breast cancer, support further clinical evaluations to determine an optimal role for the combination of capecitabine and paclitaxel in breast cancer and other relevant malignancies. PMID- 11432618 TI - Evaluation of HER2, p53, bcl-2, topoisomerase II-alpha, heat shock proteins 27 and 70 in primary breast cancer and metastatic ipsilateral axillary lymph nodes. AB - BACKGROUND: Breast cancer is a heterogeneous disease. Predictive biological markers (BM) of responsiveness to therapy need to be identified. Evaluation of BM is mainly done at the primary site. However, in the adjuvant therapy of breast cancer, the main goal is control of micrometastases. It is still unknown whether heterogeneity in the expression of BM between the primary site and its micrometastases exists. OBJECTIVE: To evaluate the expression of some BM with potential predictive value from the primary breast cancer site and metastatic ipsilateral axillary lymph nodes. PATIENTS AND METHODS: Focality (percentage of positive cells) and intensity staining scores were evaluated for each marker. Freshly cut sections (4 microm) from embedded blocks of breast cancer fixed in formalin or bouin were put onto superfrost slides (Menzel-Glaser). Protein expression was evaluated immunohistochemically (IHC) using monoclonal antibodies against: topo II-alpha (clone KiS1, 1 microg/ml, Roche) with a trypsine pre treatment (P); HSP27 (clone G3.1, 1/60, Biogenex), HSP70 (clone BRM.22, 1/80, Biogenex) and HER2 (clone CB11, 1/40, Novocastra; without P); p53 (clone D07, 1/750, Dako) and bcl-2 (clone 124, 1/60, Dako) with citrate buffer as P. RESULTS: Overall, the percentage of discordant marker status in the primary tumour and its metastatic lymph nodes was 2% for HER2, 6% for p53, 15% for bcl-2, 19% for topoisomerase II-alpha, 24% for HSP27 and 30% for HSP70. For the subgroup of patients with positive BM in the primary tumour, the percentage of discordance was 6% for HER2, 7% for p53, 14% for bcl-2, 19% for HSP70, 21% for topoisomerase II-alpha and 36% for HSP27. For the subgroup of patients with positive BM in the lymph nodes, the percentage of discordance was 9% for bcl-2, 15% for HER2 and p53, 21% for topoisomerase II-alpha, 22% for HSP27 and 25% for HSP70. CONCLUSIONS: 1) No biological marker had 100% concordant results. 2) Although some discordant cases might be explained by the limitations of the IHC technique, future studies aiming to evaluate the predictive value of BM in the adjuvant therapy of breast cancer should take into account a possible difference in BM expression between the primary and the metastatic sites. PMID- 11432619 TI - Retrospective study of the prognostic role of serum thymidine kinase level in CLL patients with active disease treated with fludarabine. AB - BACKGROUND: Previous studies have shown that the serum thymidine kinase (TK) level can be used to determine prognosis in patients with lymphoproliferative diseases, but mainly those with multiple myeloma and non-Hodgkin's lymphoma. In patients with chronic lymphocytic leukemia (CLL), TK levels may provide prognostic information independent of stage and other prognostic factors, but it is still unclear whether they can be used to predict the response to treatment and length of survival. PATIENTS AND METHODS: To determine whether TK levels can be used to predict response and survival, we retrospectively examined the serum TK level in 188 previously treated and untreated patients with active or advanced CLL who were then treated with fludarabine alone or in combination with prednisone. The correlation of the TK level with other prognostic features and with outcome was then assessed. RESULTS: Serum TK levels were elevated in 92% of the patients, and the levels proved to associate with previous treatment, stage of disease, and other tumor-burden related features (i.e., white blood cell counts, absolute lymphocyte count, bone marrow cellularity). The levels were also directly associated with indicators of tumor cell turnover (i.e., beta2 microglobulin and lactate dehydrogenase levels). Of particular importance, we found that the TK level was a significant prognostic indicator of both response to treatment and survival. Specifically, 83% of patients with a TK level of < 10 U/L responded (complete and partial response) to treatment with fludarabine, whereas only 45% of patients with a TK level of > or = 10 U/l responded to treatment (P < 0.01). This difference was maintained when we separately analyzed untreated and previously treated patients, and in patients divided according to the Binet stage. The TK level also added prognostic information about response to a predictive model based on the hemoglobin and, albumin levels and the extent of prior treatment. Of further importance, the median survival rate in patients with a TK level of < 10 U/l was 65%, as opposed to a rate of 22% in patients with a TK level of > or = 10 U/l (P = 0.000). CONCLUSIONS: The serum TK level in CLL patients provides useful prognostic information regarding both response to therapy and length of survival and should be used in planning appropriate therapy. In particular, patients with a TK level of > or = 10 U/l have a poor prognosis and should be considered for aggressive treatment. PMID- 11432620 TI - Phase I trial of gemcitabine and CPT-11 given weekly for four weeks every six weeks. AB - BACKGROUND: Our previous studies have shown that the in vitro cytotoxicity of gemcitabine and SN-38, the active metabolite of irinotecan (CPT-11), is synergistic in human tumor cell lines. PATIENTS AND METHODS: Twenty-four patients with solid tumors, refractory to standard chemotherapy or for whom no effective therapy existed (age range 31-74; 7 female, 17 male; ECOG PS 0 = 12, 1 = 11, 2 = 1), received gemcitabine and CPT-11 weekly for four weeks out of every six weeks. Fifty courses of treatment (median 2, range 1-8) were given through five dose levels of gemcitabine/CPT-11 (600/75, 800/75, 800/100, 1000/100, 1000/125 mg/m2). RESULTS: Grade 3 and 4 neutropenia occurred in eight and two patients, respectively. Grade 3 and 4 thrombocytopenia occurred in one and three patients, respectively. Hematologic toxicity resulted in > or = 2 missed doses of treatment in two out of six patients and was therefore dose limiting at gemcitabine 1000 mg/m2 and CPT-11 125 mg/m2. Grade 3 and 4 diarrhea occurred in two and one patients, respectively. Other moderate non-hematologic toxicities included alopecia, anorexia, fatigue, nausea, vomiting, and weight loss. CONCLUSIONS: The maximum tolerated dose for this study recommended for phase II testing is gemcitabine 1000 mg/m2 and CPT-11 100 mg/m2. A partial response was seen in transitional cell carcinoma. PMID- 11432621 TI - The impact of docetaxel, estramustine, and low dose hydrocortisone on the quality of life of men with hormone refractory prostate cancer and their partners: a feasibility study. AB - OBJECTIVES: The quality of life (QoL) of 44 men with HRPC and 37 partners (primary caregivers, most residing with the patient) was assessed in a multicenter Phase II trial of docetaxel, estramustine and low dose hydrocortisone (CALGB 9780). A secondary objective was to test the feasibility of assessing partners' QoL in a cooperative group setting. PATIENTS AND METHODS: Patients and partners were separately interviewed by telephone at baseline, two, four and six months by a single trained research interviewer. Patients' QoL was measured by the FACT-P, Mental Health Inventory-17 (MHI-17), Brief Pain Inventory (BPI), a two-day log of pain medications, and the OARS for co-morbid conditions. Partners' QoL was measured by the MHI-17, Caregiver Burden Interview, and co-morbid conditions. RESULTS: The QoL study refusal rates were low for patients (4%) and partners (3%). Although patients tended to experience greater treatment side effects in the first two months (FACT Physical Well-Being item, P = 0.057), their cancer-specific emotions (e.g., worrying about worsening health) significantly improved at two and four months (FACT-Emotional Well-Being, P = 0.003, P = 0.03, respectively), as did their prostate cancer-specific physical problems (e.g., urination, pain), at two and four months (FACT-P, P = 0.001, P = 0.005, respectively). Partners' anxiety significantly decreased over time (MHI, P < 0.05). Patients' quality of life at two months was significantly related to their clinical response (FACT-P total and prostate cancer-specific problems, P < 0.05), and their clinical response was significantly related to a decrease in their partners' anxiety at two months (MHI, P < 0.05). CONCLUSIONS: Despite feeling worse from side effects, patients' prostate cancer-specific problems and emotional state significantly improved in the first four months of treatment. With treatment significantly affecting both patients' and partners' lives. and the successful assessment of partners' QoL, QoL of both patients and partners could be used as important endpoints in selected clinical trials. PMID- 11432622 TI - Systematic esophageal endoscopy screening in patients previously treated for head and neck squamous-cell carcinoma. AB - BACKGROUND: An attempt was made to improve metachronous oesophageal cancer prognosis through bi-annual systematic esophageal endoscopy screening in patients treated for head and neck cancer. PATIENTS AND METHODS: Bi-annual esophageal endoscopy, without a staining procedure, was performed in 1560 patients from 1987 to 1997. The distribution of previous head and neck cancer was oral cavity (20%), oropharynx (30%), hypopharynx (34%), and larynx (16%). All patients had initial panendoscopic inspection before HNSCC treatment. Esophageal tumors were considered to be second synchronous primaries when discovered within the first six months of initial tumor diagnosis. RESULTS: Fifty metachronous esophageal asymptomatic cancers (42 T1 and 7 in situ carcinomas) were diagnosed by endoscopy. The median time between the HNC and the esophageal carcinoma was 43 months (7-137 months). Metachronous esophageal carcinoma was discovered in 2.6% of patients with oral cavity tumor, 5.7% of patients with oropharynx tumor, 2.3% of patients with hypopharynx tumor, and 1.7% of patients with larynx tumor. Causes of death were: 41.1% related to esophageal tumor with tumor progression, metastatic evolution, or treatment toxicity; 28.9% related to non malignant causes; 26.6% related to a cancer that was not of esophageal origin. CONCLUSIONS: Over a 10-year period, systematic bi-annual esophageal endoscopy uncovered metachronous esophageal tumors in 3.2% of 1560 patients originally treated for head and neck carcinoma, developing in a median time of 47 months. Patients with initial oropharyngeal tumors had a significantly higher risk of metachronous esophageal SCC, compared to the other tumor sites (P < 0.02 with Fisher exact test). Given the elevated death rate not related to the esophageal cancer and the median survival of 16 months, any potential benefit from this time-consuming procedure is debatable. PMID- 11432623 TI - Superiority of high-dose platinum (cisplatin and carboplatin) compared to carboplatin alone in combination chemotherapy for small-cell lung carcinoma: a prospective randomised trial of 280 consecutive patients. AB - PURPOSE: A prospective randomized trial in small-cell lung cancer (SCLC) was performed to determine if intensification of the platinum dose by giving cisplatin and carboplatin in combination to patients with SCLC yields higher response rates and survival, than carboplatin alone in a combination chemotherapy regimen. PATIENTS AND METHODS: Between September 1992 and October 1997, 280 patients were included in a two armed prospective randomized trial, stratified by stage of disease, LDH and performance status. The treatment was in arm A: three courses induction chemotherapy with carboplatin (AUC = 4, day 1), cisplatin (35 mg/m2, days 2 and 3), teniposide (50 mg/m2, day 1-5), vincristine (1.3 mg/m2, day 1) every four weeks, followed by cyclophosphamide (3 g/m2, day 84), 4-epirubicin (4-epidoxorubicin) (150 mg/m2, day 112), and finally one course cisplatin, carboplatin, teniposide and vincristine, (days 140-144). Arm B also comprised a total of six courses, identical to those in arm A except for omission of cisplatin. RESULTS: There were no significant differences in the overall treatment outcome for A vs. B, in terms of response rates (72% in both arms), complete response rates (40% and 34%, respectively), or median survival (314 days and 294 days, respectively). However, for patients with limited disease both the CR rate (54% vs. 37%, P < 0.05), overall survival (log-rank test, P < 0.05), and the two-year survival rate (11% vs. 6%, P < 0.05) were higher in the high-dose platinum arm compared to the carboplatin alone arm. CONCLUSIONS: The intensification of platinum dose (cisplatin plus carboplatin) in combination chemotherapy significantly increased the complete response rate, overall survival and number of two-year survivors among SCLC patients with limited disease compared to combination therapy with carboplatin alone, suggesting that a more aggressive treatment to this category of patients is worthwhile, while no difference in treatment outcome was observed for patients with extensive disease. PMID- 11432624 TI - Phase I trial of simultaneously administered GM-CSF and IL-6 in patients with renal-cell carcinoma: clinical and laboratory effects. AB - BACKGROUND: Metastatic renal-cell carcinoma is a neoplasm that is minimally responsive to cytotoxic chemotherapy. Tumor regression following therapy with cytokines such as interferon alpha and or interleukin-2 is seen in selected subsets of patients. Investigations with other immunomodulatory cytokines, such as GM-CSF and IL-6 are therefore of interest. PATIENTS AND METHODS: A phase I trial of concomitantly administered granulocyte macrophage-colony stimulating factor (3.0 mcg/kg/day s.c. d1-14) and escalating doses of interleukin-6 (1.0, 5.0 or 10.0 microg/kg/day d1-14) was conducted in patients with metastatic renal cell carcinoma to explore the toxicity of the combination and its hematologic effects. RESULTS: The most common side effects seen were fever, fatigue and arthralgias. Dose limiting toxicity included thrombocytosis and hyperbilirubinemia in patients receiving 10 microg/kg/day of IL-6. The hematologic effects of IL-6 and GM-CSF included leukocytoses and thrombocytosis, with increases in peripheral blood progenitors (BFU-E, CFU-GM, and CFU-GEMM). Evidence of platelet activation demonstrated by increased platelet expression of CD62 was found. No clinical responses were observed. CONCLUSIONS: The combination of IL-6 and GM-CSF has pleotropic hematologic effects. Further studies with this combination for the treatment of renal-cell carcinoma are not recommended. PMID- 11432625 TI - Phase I study of BBR 2778, a new aza-anthracenedione, in advanced or refractory non-Hodgkin's lymphoma. AB - BACKGROUND: BBR 2778 is a novel aza-anthracenedione showing no cardiotoxicity and superior activity compared to doxorubicin and mitoxantrone in animal models. Objectives of this phase I study included the determination of the maximum tolerated dose (MTD), the dose limiting toxicity (DLT), clinical pharmacokinetics (PK), and antitumor activity. Patients with relapsed or refractory, advanced non Hodgkin's lymphoma were included. PATIENTS AND METHODS: Patients were treated with a q1w x 3 schedule on the basis of a modified Fibonacci dose escalation method. Seven groups with a total of twenty-six patients were treated at dosages of 5, 10, 16.5, 25, 34, 56 or 84 mg/m2/w, respectively. RESULTS: DLT was observed on the seventh dose level with neutropenia WHO grade 4 in three of six patients. Pharmacokinetic analysis showed a large volume of distribution (13.5-17.5 l/kg), a high plasma clearance (0.65-1.74 l/h/kg) and a long elimination half-life (14.7 31.9 h). Tumor response included three complete remissions and two partial remissions. CONCLUSIONS: Neutropenia is the DLT of the new aza-anthracenedione BBR 2778. The recommend dose is 84 mg/m2 in a q1w x 3 schedule. PK data are consistent with a linear kinetic of BBR 2778 comparable to mitoxantrone. This new drug shows promising activity in intensively pretreated patients with relapsed or refractory NHL. Based on this results, phase II studies with this new compound are underway. PMID- 11432626 TI - Phase I study of a weekly schedule of oxaliplatin, high-dose leucovorin, and infusional fluorouracil in pretreated patients with advanced colorectal cancer. AB - PURPOSE: To identify the maximum tolerated dose (MTD) and dose limiting toxicities (DLT) of oxaliplatin (L-OHP) given on a weekly schedule including fixed doses of leucovorin (LV) and infusional 5-fluorouracil (5-FU), to define the toxicity profile of this regimen and to find preliminary evidence of its activity in pretreated patients with metastatic colorectal cancer (MCRC). PATIENTS AND METHODS: Twenty-one patients with progressive disease, treated with fluoropyrimidines and with histologically measurable MCRC entered into this phase I study. Fixed doses of LV (500 mg/m2) followed by a 48-hour 5-FU 2600 mg/m2 infusion (5-FU48h) were administered with escalating doses of L-OHP, starting from 60 mg/m2 and with stepwise increments of 5 mg/m2. No intra-patient dose escalation was allowed. Treatment was given once a week for four consecutive weeks, followed by a one-week rest period. RESULTS: Three dose levels were tested. The MTD was L-OHP 70 mg/m2 since two of the three patients showed dose limiting diarrhea and the third developed neutropenia during the first cycle of chemotherapy. Most patients complained of mild peripheral sensitive neurotoxicity, which was related to the cumulative dose of L-OHP. Treatment delays were necessary for a total of 42 cases, but only in II of 42 after the pre arranged 10% dose reduction of 5-FU (2300 mg/m2). Sixteen patients were evaluable for response: seven (33%; 95% confidence interval (CI): 14.6%-57.0%) were considered to show a major response (one complete), six showed a stable disease, and in addition progressive disease was observed in three patients. CONCLUSIONS: Our results show that L-OHP, LV and 5-FU can be administered safely and repetitively using a weekly schedule. Diarrhea and neutropenia are the DLT of this regimen. Its activity and its manageable toxicity profile deserve further evaluation in chemotherapy-naive MCRC patients. The doses recommended for phase II trials are: L-OHP 65 mg/m2, LV 500 mg/m2 and 5-FU48h 2300 mg/m2 infusion given on a weekly-times-four schedule followed by a one-week rest period. PMID- 11432627 TI - Phase II trial of bimonthly leucovorin, 5-fluorouracil and gemcitabine for advanced pancreatic adenocarcinoma (FOLFUGEM). AB - BACKGROUND: Gemcitabine alone or 5-fluorouracil (5-FU) according to several schedules are used for palliation of metastatic and locally advanced (LA) pancreatic adenocarcinoma. This study was designed to test the efficacy of the leucovorin 5-FU and gemcitabine combination. PATIENTS AND METHODS: This phase II trial combined a simplified bimonthly LV5FU2 with gemcitabine: leucovorin 400 mg/m2 in a two-hour infusion, followed by 5-fluorouracil 400 mg/m2 bolus and 2 or 3 g/m2 continuous infusion over 46 hours; gemcitabine 1 g/m2 was infused over 30 min on day 3 after 5-FU. Treatment was repeated every two weeks. Gemcitabine dose could be increased (250 mg/m2 every two cycles up to 1500 mg/m2) in the absence of NCI-CTC toxicity > 2. RESULTS: Among the 62 patients included in this study, 22 had LA and 40 had metastatic disease. Objective response rate for the 54 patients with measurable disease was 25.9% (95% confidence interval (CI): 14% 37.8%) and 22.6% (95% CI: 12%-33.2%) in the intent-to-treat population: the clinical benefit rate for the 59 assessable patients was 49.2%. Median progression-free survival and median overall survival were 4.8 and 9 months, respectively, with 32.3% of patients alive at 1 year. The most frequent toxicity (grade 3-4) was neutropenia (56.5%) usually asymptomatic (1.1% febrile neutropenia), but requiring decreases of 5-FU and gemcitabine doses. Unexpected complete alopecia occurred in 97% of patients. CONCLUSIONS: Palliative effects, response rate and survival observed in this multicenter study seem to be superior to those obtained with gemcitabine or 5-FU alone, despite a limiting hematological toxicity. PMID- 11432628 TI - A pilot study of chronomodulated infusional 5-fluorouracil chemoradiation for pancreatic cancer. AB - BACKGROUND: Dose limiting acute toxicity from chemoradiation for pancreatic cancer occurs in 15% -20% of patients treated with post-operative adjuvant therapy. Reported here is a pilot study using chronomodulated infusional 5 fluorouracil (5-FU) chemoradiation (CIC) for pancreatic cancer, a treatment designed to reduce normal tissue toxicity and maintain efficacy, with specific evaluation of acute and late morbidity, patterns of disease progression, and survival. PATIENTS AND METHODS: Twenty-three patients with adenocarcinoma of the pancreas were treated with 5-FU CIC between January 1997 and September 1999. The median age was 64, and there were 9 males and 14 females. Six patients were considered unresectable and seventeen others were treated post-operatively. The median external beam irradiation dose was 50.4 Gy. 5-FU infusion was given five days per week (300 mg/m2/d) and the median total dose was 8.4 g/m2. The chronomodulated 5-FU infusion consists of a low basal infusion rate for 16 hours followed by an eight-hour escalating-deescalating infusion peaking at 10 p.m. All patients were followed from the time of initial diagnosis until last follow-up or death; the median follow-up was 16 months. RESULTS: No RTOG grade 3 or 4 hematologic toxicity occurred. Twelve of seventeen patients treated postoperatively have been controlled locally, and seven patients have no evidence of disease. The median survival is 28 months and one-year actuarial survival is 88% in the group of resected patients. The 6 patients treated for unresectable disease have a median survival of 13 months. CONCLUSIONS: Acute toxicity of 5-FU CIC appears to be less frequent and less severe than that reported with flat infusional or bolus 5-FU based chemoradiation used for adjuvant post-operative therapy for pancreatic cancer. This method may warrant further examination, as it may be attractive for the elderly or those who cannot tolerate the toxicity associated with standard post-operative treatment protocols. PMID- 11432629 TI - Estrogen-receptor-directed neoadjuvant therapy for breast cancer: results of a randomised trial using formestane and methotrexate, mitozantrone and mitomycin C (MMM) chemotherapy. AB - BACKGROUND: We wanted to determine whether neoadjuvant systemic chemoendocrine therapy guided by the estrogen receptor (ER) status of the primary breast cancer, followed by conventional surgery and/or radiotherapy, reduces local and distant recurrence and improves survival compared with adjuvant treatment given conventionally postoperatively. PATIENTS AND METHODS: Two hundred ten patients with primary breast cancer (T1-T4, N0, N1-2) were randomised to receive treatment with neoadjuvant chemoendocrine therapy or conventional post-operative chemoendocrine therapy. Systemic therapy was based on the estrogen receptor (ER) status of the primary tumour obtained by trucut core biopsy. ER-negative patients received MMM chemotherapy (methotrexate (30 mg/m2), mitozantrone (7 mg/m2) and mitomycin (7 mg/m2) three-weekly for three months and ER-positive patients who were premenopausal received goserelin (3.75 mg monthly), and post menopausal women formestane (250 mg every two weeks) over three months. RESULTS: With a minimum of five years follow-up, there is no evidence of any survival benefit from the pretreatment neoadjuvant therapy regimen, with five year overall survival being 79% +/- 4.7% (neoadjuvant) and 87% +/- 3.4% (adjuvant). Similarly, there was no apparent benefit in terms of disease-free survival. There was, however, a significant reduction in the incidence of distant metastases in responders (4 of 51; 8%) compared with non-responders (17 of 49; 35%) (P < 0.01). There was a reduction in the need for surgery in responding patients with T1 and T2 tumours, since 10 of 74 (14%) had no detectable residual tumour, without any apparent increase in the risk of local or distant recurrence. CONCLUSION: In this study neoadjuvant treatment with endocrine or chemotherapy provided no obvious survival benefit to women with breast cancer. However, it does allow avoidance of surgery in some cases. Also, the patients whose tumours respond to neoadjuvant systemic therapy have a lower incidence of distant metastases after five year follow-up compared to those whose tumours fail to respond. PMID- 11432630 TI - High-dose chemotherapy for ovarian carcinoma: long-term results from the Solid Tumour Registry of the European Group for Blood and Marrow Transplantation (EBMT). AB - PURPOSE: to determine the outcome of epithelial ovarian cancer in patients registered with the European Group for Blood and Marrow Transplantation (EBMT). PATIENTS AND METHODS: A retrospective analysis was performed on 254 patients with advanced or recurrent disease, median age 46 years (14-63) from 39 centres treated between 1982 and 1996. Only 25% of patients were known to have no or microscopic disease after initial surgery; in approximately 20% the disease status was unknown, the remainder had macroscopic disease. RESULTS: One hundred five patients received high-dose chemotherapy in complete or very good partial remission, twenty-seven in second remission and the remainder in the presence of residual disease. Most received melphalan or carboplatin, or a combination (86%) supported by autologous bone marrow or peripheral blood stem cells. The survival of patients treated in remission was significantly better than in other groups (median 33 vs. 14 months; P = 0.0001). The durability of remission was longer after transplantation in first remission than in second remission (median disease free survival 18 vs. 9 months; P = 0.005). With a median follow-up of 76 months from diagnosis the median disease-free and overall survival in stage III disease transplanted in remission is 42 and 59 months and for stage IV disease 26 and 40 months. CONCLUSIONS: High-dose chemotherapy has a potential benefit for patients in remission. The results support the conduct of randomised studies to determine whether there is a real value from this treatment. PMID- 11432631 TI - Phase I trial of sequential administration of raltitrexed (Tomudex) and 5-iodo-2' deoxyuridine (IdUrd). AB - Raltitrexed (Tomudex) is a specific inhibitor of thymidylate synthase with clinical activity in colorectal cancer. The combination of raltitrexed and 5-iodo 2'-deoxyuridine (IdUrd, a cytotoxic pyrimidine analog) resulted in increased IdUrd incorporation into DNA and exhibited in vitro synergism against colon and bladder human carcinoma cell lines. We designed a phase I trial to determine the MTD, pharmacokinetics, and biologic effects of escalating doses of the combination of IdUrd given as a 24-hour infusion after a raltitrexed 15-minute infusion every three weeks. Thirty-four patients received 95 courses of raltitrexed and IdUrd at doses ranging from raltitrexed 1 mg/m2 and IdUrd 750 mg/m2 to raltitrexed 2.5 mg/m2 and IdUrd 10,400 mg/m2. The median number of cycles administered was 2 (range 1-10). Dose limiting hematologic toxicity occurred at doses of raltitrexed 2.5 mg/m2 and IdUrd 10,400 mg/m2. In addition, we determined the mean plasma concentrations C(SS) of IdUrd, the iodouracil level at 22 hours and the IdUrd clearance. Raltitrexed did not appear to affect the pharmacokinetics of IdUrd in the dose range tested. The recommended phase II dose is raltitrexed 2 mg/m2 and IdUrd 10,400 mg/m2 repeated every three weeks. Evidence of potential antitumor activity was observed: 1 patient (with colon cancer) had a partial response while 15 others had stable disease. PMID- 11432632 TI - Second-line treatment with oxaliplatin + raltitrexed in patients with advanced colorectal cancer failing fluoropyrimidine/leucovorin-based chemotherapy. AB - BACKGROUND: To evaluate the efficacy and tolerance of combined raltitrexed and oxaliplatin in patients with advanced colorectal cancer pretreated with fluoropyrimidine leucovorin-based chemotherapy. PATIENTS AND METHODS: Thirty-six patients with metastatic colorectal cancer, who progressed while receiving or within six months after withholding palliative chemotherapy with fluoropyrimidines leucovorin +/- irinotecan, participated in this study. Treatment consisted of oxaliplatin 130 mg/m2 and raltitrexed 3.0 mg/m2 both given on day 1 every three weeks for a total of eight courses unless prior evidence of progressive disease. RESULTS: The overall objective response rate was 33.3% for all 36 evaluable patients (95% confidence interval (CI): 18.6%-51%). Seventeen additional patients (47.2%) had stable disease, and only seven (19.5%) progressed. The median progression-free survival was 6.5 months (range 1.2-14.0). After a median follow-up time of 12 months, 23 patients (63.8%) are still alive. The tolerance of treatment was acceptable with only 8 of 36 patients (22%) experiencing grade 3 or 4 neutropenia. Grade 3 non-haematological adverse reactions included peripheral sensory neuropathy in three, asthenia in one, diarrhea in two, and clinically insignificant increase in serum transaminases in two patients, respectively. CONCLUSIONS: Our data suggest that the combination of oxaliplatin and raltitrexed has substantial antitumour activity in patients with progressive fluoropyrimidine leucovorin + irinotecan pretreated colorectal cancer. Because of its favorable toxicity profile and convenient three-weekly outpatient administration schedule, further evaluation of this regimen seems warranted. PMID- 11432633 TI - Rectal metastases from lobular carcinoma of the breast: report of a case and literature review. AB - Metastatic involvement of the gastrointestinal (GI) tract secondary to breast cancer is rare. Reported herein is the case of a 74-year-old woman with metastatic lobular breast carcinoma to the rectum presenting with obstruction. The breast tumour was diagnosed nine years prior to the presentation of rectal metastases. Endoscopy was repeated twice until a diagnosis was established. Examination of endoscopy material revealed infiltration of the rectum by malignant signet ring cells identical to those of the primary breast tumour. The patient did not respond to chemotherapy and underwent laparotomy with a defunctioning colostomy. Literature review revealed only a few more cases of metastatic breast carcinoma to the rectum. Awareness of this condition may lead to accurate diagnosis and early initiation of systemic treatment, thus avoiding surgical intervention. PMID- 11432634 TI - Pitfalls in imaging Hodgkin's disease with computed tomography and positron emission tomography using fluorine-18-fluorodeoxyglucose. AB - We report on a patient with Hodgkin's disease who presented with hypodense splenic lesions and corresponding increased glucose metabolism in FDG-PET imaging, four months after completion of initial treatment, suggestive of early relapse. Serological testing for toxoplasma gondii, however, showed evidence of a recently reactivated or newly acquired infection. Three weeks after immediate antibiotic treatment with Daraprime and Sulfadiazin, the splenic lesions had completely resolved. Additionally, serological titers for toxoplasma gondii were normalized and whole body FDG-PET imaging showed no metabolic activity. Although the positive predictive value of PET imaging to indicate lymphoma is reported to be higher than CT, hypermetabolic lesions are not specific for malignant tissue. Whereas benign tumors typically show low glucose metabolism, activated granulocytes and macrophages may display significantly increased glucose consumption. In conclusion, our case report shows that although therapeutic decisions are often based on the results of imaging modalities, the taking of a detailed history and the acquisition of histological confirmation of the suspected lymphoma relapse are also advisable where possible. Cellular immunodeficiency can result in severe infections even in patients with intermediate stage Hodgkin's lymphoma in remission after combined modality treatment. Therefore, despite the high sensitivity of FDG-PET imaging for the detection of recurrent lymphoma, the differential diagnosis of infectious lesions should be kept in mind, in particular in immunocompromised patients. PMID- 11432635 TI - Will the sentinel lymph node (SLN) stand a second time--SLN biopsy in breast cancer patients with isolated local recurrence following breast conserving therapy and previous SLN procedure. PMID- 11432636 TI - Coronary spasm induced by capecitabine. PMID- 11432637 TI - Bone scan had no role in the staging of 765 consecutive operable T(1-2)N(0-1) breast cancer patients without skeletal symptoms. PMID- 11432638 TI - Immunohistochemical distribution of cytochrome P4501A in larvae and fingerlings of the Siberian sturgeon, Acipenser baeri. AB - In this paper we investigate by means of immunohistochemistry, the tissue distribution of constitutive cytochrome P4501A (CYPIA), from hatching until 30 days posthatching in developing Siberian sturgeon, Acipenser baeri. For this purpose, a polyclonal (BN-1) antiserum developed against a conservative sequence of piscine CYP1A and a monoclonal (C10-7) antiserum directed against cod CYP1A were used on paraffin-embedded samples. From hatching onwards, distinct CYP1A immunoreactivity was distinctly observed in the following tissues and cells: envelope of oil droplets, matrix and syncytium of the yolk-sac, sinusoids, biliary epithelial cells and hepatocytes. In the digestive tract, buccopharyngeal, oesophageal, gastric and intestinal epithelia, as well as the cytoplasm and brush border of enterocytes were CYP1A-positive. Interestingly, gastric glands and melanin-plug present within lumen of the digestive system were strongly immunoreactive. Kidney (epithelia of renal tubules), gills (pillar and endothelial cells), skin (epithelial cells), muscle fibres of heart and eye (retina) were positive. In brain, we observed a strong CYP1A staining in the developing telencephalon and especially in olfactory system, as well as in those nerve fibres running ventrally toward the posterior brain. A strong CYP1A staining was observed in vascular endothelia of all organs/tissues, especially in the liver. In general, the intensity of CYP1A immunostaining increased during larval development, suggesting besides its known metabolic function (endogenous and/or exogenous), a possible participation of this heme-protein in control of cell division, regulation of growth and differentiation. PMID- 11432639 TI - Tenascin immunoreactivity in the large bowel and the liver in patients with colorectal cancer. AB - The expression of tenascin in colorectal tumours and liver was investigated in 30 patients with colorectal adenocarcinomas. Tissue samples were immersion-fixed in 4% paraformaldehyde solution. Free-floating cryostat sections were incubated with monoclonal antibody against tenascin, and examined by light and electron microscopy. Tenascin immunostaining was positive in sub-basement membrane zones and in newly-formed connective tissue of the primary tumour and perisinusoidally in the liver. The immunoreactivity in the sub-basement membrane zones of tumour glands in well- and moderately-differentiated tumours was more intensely expressed compared to that in poorly-differentiated tumours (p = 0.007 and p = 0.001 respectively, chi2-test). Perisinusoidal tenascin deposition was more often detected in the liver of patients with well-differentiated tumours (p = 0.006, chi2-test). The presence of metastases was accompanied by low tenascin deposition (p < 0.005, Fisher's exact test). Ultrastructurally tenascin deposits were observed around single tumour cells and glands in the primary tumours, and close to hepatic stellate cells in the liver. Finally, the role of tenascin deposition in the stimulation of tumour cell proliferation and mobility is discussed. PMID- 11432640 TI - Coexistence of neuropeptides and their possible relation to neuritic regeneration in primary cultures of magnocellular neurons isolated from adult rat supraoptic nuclei. AB - The coexistence of vasopressin (VP), oxytocin (OXY), galanin (GAL) and cholecystokinin (CCK) and the synthesis of GAL and CCK during neuritic regeneration was investigated in cultured magnocellular neurons, isolated from adult rat supraoptic nuclei. Double-labelling immunofluorescence was performed after 7 days of culture using primary antibodies for VP, OXY, GAL and CCK (paired in all possible combinations) and secondary antibodies labelled with either fluorescein or rhodamine. Confocal laser scanning microscopy revealed the coexistence of the mentioned peptides in all possible combinations, an unexpected result considering that the only combinations observed in tissue sections are VP GAL and OXY-CCK. Freshly dispersed cells were devoid of any neuritic processes and showed a very poor immunocytochemical staining reaction for GAL and CCK. In contrast, neurons cultured for 7, 12 and 21 days showed many neurites and a strong immunoreactivity for GAL and CCK indicative of an increased synthesis of both peptides in the regenerating neurons. This increased synthetic activity is consistent with transient upregulation of these peptides observed in situ after hypophysectomy by other authors. The results suggest that the upregulation of GAL and CCK is functionally related to the neuronal regeneration processes observed during culture and that the 'uncommon' coexistences as well as the prolonged sythesis of GAL and CCK may be due to the lack of environmental inputs, which normally regulate the expression and up- and downregulation of these peptides in vivo. PMID- 11432641 TI - Targeted fluorescent probes in peroxisome function. AB - Fluorescent peptides form a new generation of analytical tools for visualizing intracellular processes and molecular interactions at the level of single cells. The peptide-based reporters combine the sensitivity of fluorescence detection with the information specificity of amino acid sequences. Recently we have succeeded in targeting a fluorescent heptapeptide (acetyl-CKGGAKL) carrying a peroxisomal targeting signal (PTS1) to peroxisomes in intact cells. The fluorophores conjugated to the PTS1-peptide were fluorescein, BODIPY and the pH sensitive SNAFL-2. When added to cells, these fluorescent peptides were internalized at 37 degrees C and typically visible in the cell after 15 min or less. Cells lacking an active peroxisomal protein import system, as in the case of Zellweger syndrome, were stained diffusely throughout the cell. Uptake of the peptide probes was not inhibited at 4 degrees C or when the cells were depleted of ATP. Under these conditions translocation to peroxisomes was blocked. This indicates that the uptake by cells is diffusion-driven and not an active process. Using the SNAFL-2-PTS1 peptide, we established by ratio-imaging that peroxisomes of human fibroblasts have an internal pH of 8.2. The concurrent pH gradient over the peroxisomal membrane was dissipated when an ionophore (CCCP) was added. In fibroblasts of chondrodysplasia punctata patients with defects in the peroxisomal import of proteins carrying a PTS2 sequence, import of the PTS1-peptide probe into peroxisomes appeared normal, but these peroxisomes have a pH of 6.8 equal to that of the cytosol. Coupling different fluorophores to the PTS1-peptide offers the possibility of determining in time and space as to how peroxisomes function in living cells. PMID- 11432642 TI - Immunohistochemical localization and mRNA detection of Rab3D and/or Rab3B in rat von Ebner's glands, parotid gland, pancreas, and liver. AB - In examining the secretory mechanism of exocrine glands, we focused on the small GTP-binding proteins, Rab3D and Rab3B, which function in the final steps of exocytosis in non-neuronal tissues. These proteins were observed in von Ebner's glands by 32P-GTP overlay. mRNA isolated from von Ebner's glands, the pancreas, parotid glands, and liver was subjected to reverse transcription PCR and probed with primers and nested primers for Rab3D and Rab3B. Rab3D was found in all three exocrine glands and the liver, while Rab3B was found in the liver. Utilizing immunofluorescence histochemistry, Rab3D was localized to hepatocytes of the liver and to secretory granules of the exocrine glands, and Rab3B to liver and pancreatic islets. Isoproterenol evoked decreases in alpha-amylase- and Rab3D labelled parotid secretory granules, and pilocarpine stimulated decreases in secretory granules labelled for lingual lipase and Rab3D from von Ebner's glands, and amylase and Rab3D from pancreas. Neither secretagogue affected Rab3B in pancreatic islets. These observed parallel decreases in response to beta adrenergic (parotid) or cholinergic (von Ebner's and pancreas) secretagogues indicate that the function of Rab3D in exocytosis in these exocrine organs is similar and that the type of secretagogue does not determine the function. PMID- 11432643 TI - Interactions between metals in rat liver and kidney: localization of metallothionein. AB - The interactions between two essential metals, Cu and Zn, and the localization and concentration of metallothionein have been studied in rat liver and kidney. Rats receiving daily intraperitoneal injections of Cu for 3 days, or Zn for 2 days, or Cu for 3 days followed by Zn for 2 days, were sacrificed 24, 72, 120 h after the final injection. Our data indicate that Cu and Zn are both good inductors of metallothionein synthesis in rat tissues. Synergism between Cu and Zn in metallothionein synthesis was also observed as indicated by immunocytochemical experiments and chemical analysis. Moreover, in rats injected with Cu followed by Zn, the localization of metallothionein and the concentrations of both metallothionein and metal differed over time according to the organs considered. In rat kidney, a delay in the excretory process was also observed and metallothionein was present 120 h after the last injection. PMID- 11432644 TI - Immunohistochemistry of atrial natriuretic peptide in brain infarction. AB - Atrial natriuretic peptide (ANP) was originally isolated from cardiac atria, and has potent natriuretic, diuretic, and vasorelaxant properties. It has been localized in neurons and astrocytes in the cerebral cortex and the white matter. We hypothesize that glial ANP may contribute to the regulation of cerebral blood flow in brain infarction. In order to elucidate this possible role, the immunohistochemistry of ANP was studied in cases of brain infarction and in other cases of brain trauma for comparison. A statistically significant increase in the number of ANP-immunoreactive glial cells (mainly astrocytes) was observed in the white matter surrounding the brain infarction compared with the intact area. No statistically significant increase in ANP-immunoreactive glial cell number was observed in the cerebral white matter from brain haemorrhage, contusion and control cases. Our results indicate that glial ANP may increase in number in brain infarction, and that it may be involved in the regulation of the cerebral blood flow in the infarcted area. PMID- 11432645 TI - Distinctive expression of extracellular matrix molecules at mRNA and protein levels during formation of cellular and acellular cementum in the rat. AB - Little is known about differential expression of extracellular matrices secreted by cementoblasts between cellular and acellular cementum. We hypothesize that cementoblasts lining acellular cementum express extracellular matrix genes differently from those lining cellular cementum, thereby forming two distinct types of extracellular matrices. To test this hypothesis, we investigated spatial and temporal gene expression of selected extracellular matrix molecules, that is type I collagen, bone sialoprotein, osteocalcin and osteopontin, during formation of both cellular and acellular cementum using in situ hybridization. In addition, their extracellularly deposited and accumulated proteins were examined immunohistochemically. The mRNA transcripts of pro-alpha1 (I) collagen were primarily localized in cementoblasts of cellular cementum and cementocytes, while those of bone sialoprotein were predominantly seen in cementoblasts lining acellular cementum. In contrast, osteocalcin was expressed by both types of cementoblasts and cementocytes and so was osteopontin but only transiently. Our immunohistochemical examination revealed that translated proteins were localized extracellularly where the genes had been expressed intracellularly. The present study demonstrated the distinctive expression of genes and proteins of the extracellular matrix molecules between cellular and acellular cementum. PMID- 11432646 TI - An overview of technological resources used in rehabilitation research with people with severe/profound and multiple disabilities. AB - PURPOSE: This paper provides an outline of technological resources used in rehabilitation research with persons with severe/profound and multiple disabilities, and discusses the practicality of such resources. METHOD: Research articles published during the last 8 years (1993-2000) are considered. With regard to each of the technological resources used in the articles (i.e. microswitches, speech output devices, computer-aided instruction systems, audio recording systems, orientation systems, mobile robot, and SIBIS), accessibility, cost and implementation issues are examined. RESULTS AND CONCLUSIONS: Some technological resources are easily accessible and affordable (i.e. microswitches, speech output devices, audio-recording systems and SIBIS). Other resources are still at the prototype level (i.e. computer-aided instruction systems, orientation systems and robot) and are much more complex and costly. In spite of this, the latter resources might still have an important role in daily contexts. PMID- 11432647 TI - The epidemiology of disability and occupation handicap resulting from major traumatic injury. AB - PURPOSE: To assess the prevalence of disability and handicap among survivors of major traumatic injury (injury severity score > 15) using a prospective population based cohort study design. The study was set in the former Yorkshire Health Region. SUBJECTS/METHODS: A cohort of 367 individuals identified as having received and survived major traumatic injury during the 12 month period October 1988-September 1989. OPCS disability scores and employment status at 5 years post injury were established through structured face-to-face interviews. RESULTS: Three hundred and four (84% response. 89% adjusted response) individuals were interviewed. Seventy-seven percent of these were male and they had a mean age (SEM) of 30.8 (1.06) years. Injuries were caused by road traffic accidents in 68% of the cases and were primarily orthopaedic and neurological in nature. At 5 years post injury 81.2% of individuals had some form of measurable disability principally relating to locomotion, behaviour, continence and intellectual functioning consistent with injury type. A third had an OPCS disability score of 5 or greater and approximately 1 in 12 were in the most severe categories of OPCS scores of 9-10 necessitating dependency on formal or informal carer assistance. Whilst five of the eight sub-scales of the SF36 showed correlation in severity proportion, general health perception and energy/vitality were higher in those with increasing disability as measured by the OPCS scale. Of those between the ages of 16-64 nearly half (49%) were not in paid employment at the time of follow up. CONCLUSION: A high prevalence of severe permanent disability, work disability and occupation handicap has been identified in a cohort of mainly young adult males following major traumatic injury resulting from road traffic accidents. Progress in accident prevention, injury reduction and the management of patients with serious injuries should be measured not only in terms of reduced mortality from such events but also in the long term disability and quality of life sequelea of survivors. PMID- 11432648 TI - Health status and quality of life of people with multiple sclerosis. AB - PURPOSE: The aim of this study was to ascertain the health status and quality of life of a community based cohort of people with multiple sclerosis. METHOD: A postal questionnaire with self-completed measures of impairment, disability, physical dependency and quality of life was sent to a random sample of 203 people with multiple sclerosis from a population register. The sample was stratified according to five disease courses. The population register is of the prevalent population of 760 people with multiple sclerosis resident in the Leeds Health Authority. The register used multiple sources of ascertainment and is prospectively maintained with new incident cases. RESULTS: The estimated mean age of people with multiple sclerosis is 46 years (SE: 0.85), and mean duration of disease is 14.4 years (SE: 0.69). Almost four in five (78 %) are female, and one in six (17%) live alone. Impairments of balance, vision and memory are common and in all cases there is little difference in the frequency between disease course groups. In contrast, impairments of bladder and bowel are more common in those with a progressive disease course. Disability is more common in those with a progressive disease course but all scores on the SF36 Physical Function scale are low and demonstrate the disabling consequences of the disease, irrespective of disease course. These consequences must contribute to the fact that over two thirds (68 %) were not employed at the time of the survey. Quality of life does not differ across disease course groups, but rather varies by age and duration. CONCLUSIONS: People with multiple sclerosis experience a range of impairments and disabilities. Those with progressive disease courses experience greater levels of impairment and disability than other groups. There is not a straightforward exchange between health status and quality of life. A measure of subjective quality of life may reflect adjustment to disease, such that, for example, the longer the duration, the older the individual, the more likely the person will report a relatively good quality of life. PMID- 11432649 TI - Phantom limb pain and residual limb pain following lower limb amputation: a descriptive analysis. AB - PURPOSE: This research aimed to develop a clearer picture of the experience of residual limb pain and phantom limb pain following a lower limb amputation and to gain a greater understanding of their relationships with physical and psychosocial variables. METHOD: One hundred and four participants completed the Trinity Amputation and Prosthesis Experience Scales (TAPES), which includes a section on each of, psychosocial issues, activity restriction, satisfaction with a prosthesis and pain (incidence, duration, level and extent of interference). RESULTS: The results showed that 48.1% of the sample experienced residual limb pain and 69.2% experienced phantom limb pain. While fewer people experienced residual limb pain, those who did, experienced it for longer periods, at a greater level of intensity and with a greater amount of interference in their daily lifestyle, than people who were experiencing phantom limb pain. The experience of residual limb pain was associated with other medical problems and low levels of Adjustment to Limitation. Phantom limb pain was associated with older age, being female, above knee amputation, causes other than congenital causes, not receiving support prior to the amputation, the experience of other medical problems, low scores on Adjustment to Limitation and high scores on Aesthetic Satisfaction with the prosthesis. CONCLUSION: These findings provide a greater understanding of the issues to be taken into consideration in the rehabilitation of people with a lower limb amputation. PMID- 11432650 TI - Comparison of subacute rehabilitative care with outpatient primary medical care. AB - PURPOSE: Prior rehabilitation outcome studies have had many weaknesses. but they gradually observe a lack of long-term benefits from inpatient care alone. The goal of this study was to measure the additive effect of outpatient, subacute rehabilitation (compared with usual outpatient primary medical care) for adults diagnosed with a disabling disorder in four major diagnostic groups (nervous, circulatory, musculoskeletal and injury). METHOD: A randomized clinical trial was conducted to determine the effects of outpatient, subacute rehabilitative care on: (1) physical function; (2) health; (3) well being; (4) family function; and (5) social support. Patients hospitalized for the first time with a disabling condition (n = 180) were provided inpatient rehabilitation and then were randomly assigned to either outpatient, subacute rehabilitation at home (n = 90) or to usual outpatient follow-up (n = 90) in which only primary care medical services were provided. To compare the two groups, univariate analyses of covariance were conducted for the outcome variables. RESULTS: The major finding of this study was that of no significant effect of the intervention on any outcome variable. CONCLUSIONS: Based on current study results, we conclude that any long term additive benefit of outpatient, subacute rehabilitation may not be detectable across disability categories and may require closer evaluation in studies with a more homogeneous population than in the current study. Providing complex follow up case management services to all clients is apparently not beneficial and might better be provided using selection criteria based on need. Future studies should determine if services are more effective when provided to those with the most unmet rehabilitative needs. Further outpatient, subacute care rehabilitation studies should address the specific needs of the patients and be adapted individually to those needs. PMID- 11432651 TI - Changing chairs: anticipating problems in prescribing wheelchairs. AB - PURPOSE: This article presents a framework for prescribing, ordering, and adapting a new wheelchair, focusing on individual, environmental and wheelchair factors that must be taken into consideration to ensure optimal function. METHOD: A review and analysis was conducted of all factors relevant to the transition to a new wheelchair. Without appropriate planning and implementation, this transition can result in unnecessary expenses, duplication of effort, and possibly even injury to the user and abandonment of the wheelchair. RESULTS: Recommendations are provided to manufacturers, therapists, technicians, users, insurers and physicians, who must work together throughout this process. To the extent feasible, the authors suggest that major changes from the previous wheelchair should be avoided, particularly for people with substantial functional limitations. Therapists and technicians must measure the user accurately, and anticipate those factors that can impede a smooth transition. Insurers and other payors must recognize that changing wheelchairs will often require substantial professional assistance, including several fittings to adjust the new chair to the needs of the user. CONCLUSION: Additional research and case reporting on outcomes of adjusting to a new wheelchair appear warranted. PMID- 11432652 TI - Bone and soft tissue integration to titanium implants with different surface topography: an experimental study in the dog. AB - The aim of the present experiment was to study the peri-implant soft and hard tissues formed at titanium implants with 2 different surface configurations and to give a topographic description of the surfaces examined. In 5 beagle dogs, the mandibular premolars were extracted. Three months later, 4 self-tapping standard implants (SI) and 4 Osseotite implants (OI) of the 3i Implant System were placed. The marginal 3 mm of the OI is turned, while the remaining part has an acid etched surface structure. Abutments were connected after 3 months. A plaque control period was initiated, and after 6 months block biopsies were obtained. From each animal 2 units of each implant type were processed and embedded in EPON. The remaining biopsies were processed for ground sectioning. The histometric measurements performed on the EPON sections revealed that the peri implant soft tissues and the marginal level of bone-to-implant contact were similar for SI and OI sites. In the ground sections, bone-to-implant contact (BIC%) and bone density assessments were made in 2 different zones. Zone I represented the contact area measured from the marginal level of bone-to-implant contact (B) to a position 4 mm above the apex of the implant, and zone II represented the apical 4 mm of the implant. For the SI sites, the BIC% was 56.1% in zone II and 58.1% in zones I + II. The corresponding figures for the 01 sites were 76.7% and 72.0%. The BIC% was significantly larger at OI than at SI sites. Bone density values were similar at the SI and OI sites. PMID- 11432653 TI - Surface characterizations of variously treated titanium materials. AB - The attachment of cells to titanium surfaces is an important phenomenon in the area of clinical implant dentistry. A major consideration in designing implants has been to produce surfaces that promote desirable responses in the cells and tissues. To achieve these requirements, the titanium implant surface can be modified in various ways. This research was designed to elucidate the relationship between surface roughness (Ra) and contact angle (theta) of various engineered titanium surfaces of commercially pure titanium, titanium-aluminum vanadium alloy (Ti-6Al-4V), and titanium-nickel (TiNi) alloy. The contact angle was measured using distilled water, 1% sodium chloride solution, human neutrophils, and osteoblast-like cells. Surface oxide crystallography was identified by transmission electron diffraction. It was found that: (1) there were no significant differences in contact angles among the 4 media; (2) for commercially pure titanium, a combined treatment (hydrofluoric acid/nitric acid/water --> sodium hydroxide --> oxidation) showed the lowest theta (10.51 degrees in water), while the surface treated with sulfuric acid showed the highest value (72.99 degrees in water); (3) for all commercially pure titanium samples, when theta is greater than 45 degrees, the contact angle increases linearly with Ra (hydrophobic nature) and the surface is covered with rutile-type oxide only, while the contact angle decreases linearly with Ra when theta is less than 45 degrees (hydrophilic nature) and the surface is covered with a mixture of rutile and anatase oxides; and (4) a similar trend was found on Ti-6Al-4V and TiNi surfaces. PMID- 11432654 TI - Lateral ridge augmentation and implant placement: an experimental study evaluating implant osseointegration in different augmentation materials in the canine mandible. AB - The present study investigated the osseointegration of dental implants with a titanium plasma-sprayed surface (TPS) in regenerated and native bone in an experimental dog study. Initially, lateral bone defects were created in the alveolar ridge on both sides of the mandible. Two months later, lateral ridge augmentation was performed with (1) autogenous corticocancellous block grafts, (2) autogenous corticocancellous block grafts and e-PTFE membrane, (3) tricalcium phosphate particles and e-PTFE membrane, or (4) canine-derived demineralized freeze-dried bone allograft particles and e-PTFE membrane. After 4 months, membranes were removed, and non-submerged titanium implants were placed in regenerated bone (test implants) and in native bone (control implants). Two months later, the animals were sacrificed and non-decalcified orofacial sections were evaluated histometrically. All implants demonstrated high percentages (59% to 75%) of bone-to-implant contact, with no significant differences across the various treatment groups. The different grafting techniques did not significantly influence the location of first bone-to-implant contact and the horizontal bone width at the most coronal bone level. PMID- 11432655 TI - Long-term results of implants treated with guided bone regeneration: a 5-year prospective study. AB - The aim of this prospective 5-year longitudinal study was to follow endosteal implants in which guided bone regeneration (GBR) was applied during implant placement. In 75 patients, defects around implants (Branemark System) were treated with Bio-Oss and Bio-Gide (112 implants). In split-mouth patients in this group, Bio-Oss and Gore-Tex were used in the second defect site (41 implants). All 75 patients had at least 1 implant that was entirely surrounded by bone and served as the control (112 implants). After placement of the definitive prostheses (single-tooth, fixed, or removable implant prostheses), patients were recalled after 6 months and then every 12 months during a 5-year observation period. The following variables were investigated: implant survival, marginal bone level (MBL), presence of plaque, peri-implant mucosal conditions, height of keratinized mucosa (KM), and marginal soft tissue level (MSTL). The cumulative implant survival rate after 5 years varied between 93% and 97% for implants treated with or without GBR. The mean MBL after 60 months was 1.83 mm for sites treated with Bio-Oss and Bio-Gide, 2.21 mm for sites treated with Bio-Oss and Gore-Tex, and 1.73 mm for the control sites. The MBL values were found to increase significantly with time and differed significantly among the treatment groups. During the observation period, KM varied between 3.16 and 3.02 mm. A slight recession of 0.1 mm was observed, and plaque was found in 15% of all sites and was associated with inflammatory symptoms of the peri-implant mucosa. It was observed that such symptoms and recession correlated more strongly with the type of restoration than with the type of treatment. This study demonstrated that implants placed with or without GBR techniques had similar survival rates after 5 years, but that bone resorption was more pronounced in sites with GBR treatment. It was assumed that the use of GBR is indeed indicated when the initial defect size is larger than 2 mm in the vertical dimension. PMID- 11432656 TI - Implant-abutment screw joint preload of 7 hex-top abutment systems. AB - This study measured the screw joint preload of the implant-abutment interface for 7 common hex-top abutment systems. Increasingly, prosthetic designs are utilizing a direct prosthetic connection to the implant, placing the implant-abutment screw joint under direct functional loads and moments. Sufficiently high screw joint preloads are required to maintain screw joint integrity and confer clinical longevity to implant prosthetic components to prevent such complications as abutment screw loosening and screw fracture. Strain-gauged abutment load cells were calibrated to measure screw joint pre-load at the implant-abutment interface. Torque delivery by electronic torque controller was varied at low- and high-speed settings. At manufacturer's recommended torques, the overall mean preload measured was 181.6 +/- 60.0 N for the Nobel Biocare Standard abutment, 291.3 +/- 41.2 N for the Nobel Biocare EsthetiCone abutment, 456.5 +/- 44.0 N for the Nobel Biocare MirusCone abutment, 369.7 +/- 32.9 N for the 3i Titanium Abutment post, 643.4 +/- 143.1 N for the Nobel Biocare CeraOne abutment, 536.3 +/ 68.6 N for the Nobel Biocare "Gold Cylinder to Fixture" abutment, and 556.9 +/- 145.6 N for the Nobel Biocare TiAdapt abutment. Analysis of variance revealed significant differences between abutment systems (P < .001) and between torque driver speed settings (P < .001). Implant-abutment screw joint preload of external-hex implants is dependent on abutment design, screw diameter, material, tightening torque, and torque controller speed. PMID- 11432657 TI - Ridge augmentation using mandibular block bone grafts: preliminary results of an ongoing prospective study. AB - The aim of the current ongoing study is to evaluate the long-term results of endosseous implants placed into autogenous bone grafts from intraoral donor sites. Patient selection for the correction of bone deficiencies was based on biomechanical and esthetic needs. Donor site selection was dependent upon the type of deficiency and the graft shape needed. Two-stage implants were placed after a healing period of 3 to 6 months, based on an assessment of the graft viability with radiographic and clinical parameters. Thus far, 118 implants have been placed in 60 patients whose alveolar ridges were deficient in height, width, or both height and width and were augmented. The patients were observed for up to 77 months. Two implant failures were encountered before implant exposure (1.7%). No further implants have been lost in function. PMID- 11432658 TI - Vascular endothelial growth factor and microvessel density around healthy and failing dental implants. AB - Inflammatory infiltrate may be important in the evolution of inflammatory processes involving peri-implant tissues. Angiogenesis is an important feature of inflammation and healing, but its role in the development and progression or in the healing of periodontal lesions has not been elucidated. Vascular endothelial growth factor (VEGF) is a potent inducer of endothelial cell proliferation. The aim of the present study was to conduct a comparative immunohistochemical evaluation of VEGF and microvessel density (MVD) in normal keratinized gingiva and in peri-implant soft tissues surrounding failing implants. Fifteen patients participated in this study. Ten biopsies were taken from healthy keratinized gingiva, and 10 were taken from peri-implant soft tissues surrounding failing non submerged implants. In healthy sites, the endothelial lining cells of the vessels always tested positive for VEGF; also, VEGF intensity was high in most cases. Stromal cells were positive for VEGF in 70% to 90% of samples. The MVD was 60.250 +/- 5.123. In peri-implantitis samples, the cells of the inflammatory infiltrate were positive for VEGF in 80% to 100% of cases, and the VEGF intensity was low in all cases. The stromal cells were positive for VEGF in 90% to 100% of cases, and in most cases the intensity was low. The MVD was 101.800 +/- 11.256. The difference in MVD between healthy sites and peri-implantitis was statistically significant (P = .0158). Expression of VEGF was lower in peri-implantitis samples, and this difference was statistically significant (P = .0373). Because of its extensive presence, VEGF is probably a factor in both the maintenance of periodontal physiology and in the progression of peri-implant inflammatory disease. PMID- 11432659 TI - Accuracy of electronic implant torque controllers following time in clinical service. AB - Tightening of the screws in implant-supported restorations has been reported to be problematic, in that if the applied torque is too low, screw loosening occurs. If the torque is too high, then screw fracture can take place. Thus, accuracy of the torque driver is of the utmost importance. This study evaluated 4 new electronic torque drivers (controls) and 10 test electronic torque drivers, which had been in clinical service for a minimum of 5 years. Torque values of the test drivers were measured and were compared with the control values using a 1-way analysis of variance. Torque delivery accuracy was measured using a technique that simulated the clinical situation. In vivo, the torque driver turns the screw until the selected tightening torque is reached. In this laboratory experiment, an implant, along with an attached abutment and abutment gold screw, was held firmly in a Tohnichi torque gauge. Calibration accuracy for the Tohnichi is +/- 3% of the scale value. During torque measurement, the gold screw turned a minimum of 180 degrees before contact was made between the screw and abutment. Three torque values (10, 20, and 32 N-cm) were evaluated, at both high- and low-speed settings. The recorded torque measurements indicated that the 10 test electronic torque drivers maintained a torque delivery accuracy equivalent to the 4 new (unused) units. Judging from the torque output values obtained from the 10 test units, the clinical use of the electronic torque driver suggests that accuracy did not change significantly over the 5-year period of clinical service. PMID- 11432660 TI - Impact of recombinant human bone morphogenetic protein-2 on residual ridge resorption after tooth extraction: an experimental study in the rat. AB - Residual ridge resorption begins following tooth extraction and continuously reduces alveolar bone volume, potentially creating a significant problem in dental implant treatment. In this study, the role of recombinant human bone morphogenetic protein-2 (rhBMP-2) in residual ridge resorption after tooth extraction was investigated. A polylactic acid/polyglycolic acid copolymer-coated gelatin sponge carrier was implanted with or without rhBMP-2 (1 microg) in the mesial root sockets after removal of maxillary first molars in male Wistar rats. Fine structural and histomorphologic analyses were conducted 3 to 84 days after implantation. Direct bone formation was first observed after 5 days on the rhBMP 2 side, which was transformed into cortical alveolar ridge with a smooth periosteal layer by 84 days, whereas the control side displayed slower healing. Bone histomorphometry revealed greater total bone area and increased bone height after 14, 28, 56, and 84 days on the rhBMP-2 side compared to the control side, and differences were significant after 14, 28, and 56 days. Larger numbers of proliferating cells and densely populated differentiating mesenchymal cells were observed on the rhBMP-2 side than on the control side in the early stage, and chondrogenesis was not observed. The findings indicate that rhBMP-2 may stimulate proliferation and differentiation of mesenchymal cells in the rat maxillary root socket to preserve cortical bone volume in the socket without any evidence of chondrogenesis. PMID- 11432661 TI - Development and treatment of retrograde peri-implantitis involving a site with a history of failed endodontic and apicoectomy procedures: a series of reports. AB - Osseointegrated implants provide predictable restorative support for crowns, restorations, prosthesis abutments, and removable dentures. Their widespread use in recent years has produced different types of complications. Retrograde peri implantitis, a lesion occurring at the periapical area of an osseointegrated implant, has recently been described. This paper presents a series of reports describing the occurrence and management of retrograde peri-implantitis involving implants replacing teeth with histories of failed endodontic and apicoectomy procedures. PMID- 11432662 TI - Immediate loading of implants with fixed restorations in the completely edentulous mandible: report of 27 patients from a private practice. AB - Numerous authors have documented clinical success in loading threaded implants at the time of implant placement when carefully controlled surgical and restorative protocols are followed. This clinical series documents the application of immediate loading techniques to fixed mandibular restorations in 27 patients who were edentulous or had non-restorable mandibular dentitions. Eighteen patients had complete conventional maxillary dentures, while 9 had natural or fixed prosthetic maxillary dentitions. Twelve different restorative dentists provided prosthetic support for these patients and used 4 different restoration types: laboratory- or office-processed, with cement or screw retention. Five to 8 threaded implants were placed in each patient. One hundred sixty-one of the 186 implants that were placed by the authors were loaded immediately using fixed provisional restorations of the various designs. More than 99% (160/161) of the immediately loaded implants and 99.5% (185/186) of all implants were clinically integrated and radiographically successful at the time of final evaluation for restoration fabrication. After final implant evaluation, no additional implant losses occurred, indicating an implant survival rate of nearly 100% over a mean of 25.0 months (range 13 to 41 months) following implant placement. While a strong preference for cement-retained restorations was apparent, all prostheses showed similar success. The data and the experience described in this report indicate that immediate loading with fixed restorations using appropriate surgical and restorative techniques can be a predictable technique for rehabilitation of the completely edentulous mandible. PMID- 11432663 TI - Evaluation of the use of iliac cancellous bone and anorganic bovine bone in the reconstruction of the atrophic maxilla with titanium mesh: a clinical and histologic investigation. AB - The present article describes a titanium mesh procedure used for bone augmentation in the treatment of severe maxillary atrophy. A mix of iliac cancellous bone and anorganic bovine bone in a 1:1 ratio is proposed for achieving the best bone quality at the time of implant placement, which is performed 5 to 6 months after the augmentation surgery. This procedure provides for 3-stage surgery using a titanium mesh (which is removed 4 to 5 months later) to retain the cancellous bone/Bio-Oss mixture. Bone specimens taken 5 to 6 months after the augmentation procedure showed bone regeneration and the presence of vessels, indicating bone vitality. PMID- 11432664 TI - Oral rehabilitation using osseointegrated implants in a patient with idiopathic torsion dystonia. AB - Idiopathic torsion dystonia is a motor syndrome characterized by dystonic movements and postures in the absence of other neurologic deficits. The condition involves prolonged spasms of muscle contraction that distort the body into typical postures. Such distortions involving the head and the neck make conventional denture use in edentulous patients very difficult. The present paper reports on a patient with idiopathic torsion dystonia who was treated with a mandibular overdenture supported by endosteal implants, which enabled the establishment of a stable occlusion and improved the dynamics of the masticatory muscles for chewing. PMID- 11432665 TI - Primary gastrointestinal B-cell lymphoma. A clincopathological and immunohistochemical study of 61 cases with an evaluation of prognostic parameters. AB - We hereby present a retrospective clinicopathological and immunohistochemical study of surgically resected primary gastrointestinal (GI) lymphoma with an analysis of parameters of potential prognostic relevance. From a larger series of 144 cases of primary GI lymphomas, we chose 61 cases with sufficient clinical follow-up (mean 60, range 1-219 months), classified either as extranodal marginal zone B-cell lymphoma of MALT type (MALT lymphoma) or diffuse large B-cell lymphoma (DLBCL), after having excluded other subtypes. In addition to conventional clinical and morphological parameters, the expression levels of Ki 67 (MIB-1), bcl-2 and p53 were evaluated for prognostic significance. Twenty-one (34.4%) cases were classified as pure low grade marginal zone B-cell lymphoma of MALT type, 12 (19.7%) cases as low grade MALT lymphoma with a high grade component (mixed type), and 28 (45.9%) cases as primary extranodal DLBCL. Most of the lymphomas (53/61; 86.9%) were localized in the stomach, 3 (4.9%) in the small bowel, 3 (4.9%) multifocal in both stomach and small intestine and 2 (3.3%) in the large bowel. MIB-1 expression in more than 30% of tumor cells was detected in 42 (68.6%), bcl-2 expression in 20 (32.8%) and p53 accumulation in more than 10% of neoplastic cells in 16 (26.2%) lymphomas. Both high Ki-67 expression and p53 accumulation were more prevalent in the DLBCL. 30 (49%) patients showed lymph node involvement at surgery, 14 (23%) patients suffered tumor recurrence, and 24 (38.5%) died during the follow-up period. Tumor recurrence occurred primarily in patients who had presented lymph node involvement (9/14, 64.3%). The 5-year survival rate was 66.1% for all patients. Important prognostic factors for overall survival were tumor stage (p < .004) and p53 accumulation (p < .05) in univariate analysis, and tumor stage in multivariate analysis (p < .001). Although p53 accumulation did not reach statistical significance in our small study group, it may be both important in the transformation of low grade MALT lymphoma and an indicator for aggressive behavior in high grade tumors. PMID- 11432666 TI - Clinicopathologic characteristics of Epstein-Barr virus-incorporated gastric cancers in Korea. AB - We evaluated the rate of Epstein-Barr virus (EBV) infection in gastric carcinomas of Korean patients and investigated the associations between EBV infection and clinicopathological characteristics, the survival rates of patients, and p53 overexpression. EBV-encoded small RNA (EBER)-in situ hybridization and immunohistochemistry for p53 protein were done in 306 consecutive gastric carcinoma cases, of which 17 (5.6%) showed EBV infection. Of these 17 EBV positive cases, one case strongly expressed p53 protein, while 98 (34%) of 285 EBV-negative cases overexpressed p53 (p < 0.05). The EBV-positive gastric carcinomas tended to have lymphoid stroma. They were mostly of the poorly differentiated type, negative for p53 immunoexpression, more prevalent in male patients, and diffuse according to Lauren's classification (p < 0.05). There was no significant difference in the survival rate for the EBV status. In conclusion, the EBV infection rate among gastric carcinomas in Korea is similar to that ascertained in other countries. An inverse correlation between EBV and p53 overexpression was disclosed. Further study is needed to find out whether or not two genetic changes could be functionally overlapping during gastric carcinogenesis. PMID- 11432667 TI - Differences in EBNA2 and LMP-1 carboxy terminal region sequences of Epstein-Barr virus type A between the tumors in a multiple cancer patient. AB - Using PCR, type A Epstein-Barr virus (EBV) infection was demonstrated in a squamous cell carcinoma of the maxilla (in a 52-year-old man) and the tongue of the same patient 18 years later (at the age of 70). Furthermore, at the age of 72, this patient developed an EBV-infected anaplastic large cell lymphoma. Analysis of the terminal regions of the EBV genome revealed a monoclonal proliferation of EBV-infected lymphoma cells. However, sequence analysis of the EBV revealed a slight difference in the EBNA2 regions between the virus-infected lymphoma and the squamous cell carcinomas. The mutations at 48991 (G-->T) and 48998 (C-->A) were demonstrated in the lymphoma. Although the squamous cell carcinoma of the tongue occurred after an interval of 18 years, the mutation site in the carcinomas was the same, 49137 (A-->G), as compared with B95-8 strain EBV EBNA2. The mutations at 48991 and at 49137 were associated with amino acid changes, Arg-->Met and Thr-->Ala, respectively, but the alteration at 48998 was a silent mutation. Thirty-bp deletion in the LMP-1 carboxy terminal region was demonstrated in the virus-infected lymphoma, but not in the squamous cell carcinomas. On the other hand, HTLV-1 proviral DNA (tax, gag and env) was not detected in the lymphoma, nor was HPV demonstrated in the squamous cell carcinomas, although Okinawa is known as an HTLV-1 and HPV prevalence region. The T-cell receptor beta gene rearrangement was demonstrated in the lymphoma, but the t(2;5) fusion transcript was not detected using PCR. Cytogenetic analysis of the lymphoma cells showed a complex hypertriploid karyotype with 76XY. The type A EBV infection might play a role in the carcinogenesis of the tumors of our patient. Interestingly, the infected virus genome sequences, the EBNA2 and LMP-1 regions, which were closely associated with carcinogenesis in the squamous cell carcinomas and the lymphoma, showed slight differences. PMID- 11432668 TI - Cell proliferation in colorectal adenocarcinomas: comparison between Ki-67 immunostaining and bromodeoxyuridine uptake detected by immunohistochemistry and flow cytometry. AB - We compared three different means of assaying tumor proliferative activity in 30 human colorectal adenocarcinomas labeled in vivo with bromodeoxyuridine (BrdUrd). The labeling indices (LI) of BrdUrd obtained both by flow cytometry (FCM) and immunohistochemistry (IH) were also compared with the labeling index of Ki-67. These methods were then related to tumor ploidy and pathological features. Flow cytometry was performed in accordance with Begg's method after intravenous infusion of BrdUrd four hours before surgery. Immunohistology was carried out on paraffin-embedded sections with monoclonal antibodies against BrdUrd and Ki-67. A positive correlation was found between BrdUrd LI obtained by both FMC and IH (p<0.0001), a finding that complies with the literature. However, we report on a correlation between Ki-67 LI and BrdUrd LIs in colorectal tumors (p=0.012). The results were valid for all tumors when they were subdivided into diploid and aneuploid groups. The labeling indices were significantly higher in the aneuploid tumor group than in the diploid group (p=0.047). No relationship between proliferation parameters and tumor stage or grade was found. To our knowledge, this is the first report on a positive correlation between tumor proliferation indices in BrdUrd LIs and Ki-67 in colorectal carcinomas. This finding validates the value of Ki-67 immunostaining, which, however, should be confirmed in a larger series under the same technical conditions. PMID- 11432669 TI - EMA: a differentiation antigen related to node metastatic capacity of breast carcinomas. AB - Epithelial membrane antigen (EMA), also known as MUC1, is a mucinous glycoprotein fixed to the luminal domain of the epithelial cell membrane of normal breast ducts. However, in breast cancer cells, it is usually dispersed in the cytoplasm. EMA staining patterns of 330 breast carcinomas were examined, and three groups formed: lineal (16%), cytoplasmic (75%), and negative (9%). Although these patterns were somewhat related to histological cancer types, this was not statistically significant. However, EMA showed statistically significant univariate relationships to tumor grade, tumor size, estrogen and progesterone receptors, and nodal stage. Logistic regression analysis showed that among these variables, all of which were univariately related to node metastasis, only tumor size and EMA were independent nodal stage predictors. A combined analysis of these two factors revealed that the statistical probability of a tumor metastasizing to four or more nodes increased in each tumor size group from 0.9% to 12% for pT1, from 2% to 29% for pT2 and from 10% to 63% for pT3, depending on the EMA staining. The tumors showing a lineal pattern were the least metastasizing, while the EMA-negative tumors were the most. After recognizing these relationships between EMA staining patterns and other well-known differentiation markers and the lymph node metastatic capacity of carcinomas, and considering the results obtained by others on survival, one might conclude that EMA is both a differentiation marker and a histological prognostic factor. PMID- 11432670 TI - Distribution of epidermal growth factor receptor protein correlates with gain in chromosome 7 revealed by comparative genomic hybridization after microdissection in glioblastoma multiforme. AB - In a recent study, 23 microdissected areas of 10 glioblastoma multiforme (GBM) were investigated for quantitative genomic aberrations using comparative genomic hybridization (CGH). To validate the chromosomal aberrations, as revealed by CGH after microdissection, parallel tissue sections were stained immunohistochemically with an antibody that detects both wild-type epidermal growth factor receptor (EGFR) and the deletion mutant form of the receptor (EGFRvIII). Immunostaining was correlated with CGH data of chromosome 7, because chromosome 7 is the most frequently aberrant chromosome in GBM (here four of 10 tumors), and this aberration often indicates an abnormality of EGFR. Nine of nine areas that showed gain in or amplification (2 areas) of chromosome 7 with CGH contained EGFR-immunoreactive cells. Only three of 14 areas without abnormality of chromosome 7 in CGH contained EGFR-immunoreactive cells; eleven of 14 areas were immunonegative. Our findings demonstrate a strong correlation between immunohistochemistry of EGFR and the copy numbers of chromosome 7, as revealed by CGH after microdissection in glioblastoma multiforme. PMID- 11432671 TI - Diffuse hepatic intravascular carcinomatous embolization resulting in fatal liver failure: a clinicopathologic study of 4 cases. AB - The authors report four cases in which patients died of acute or fulminant hepatic failure resulting from massive intravascular metastatic carcinomatous embolization, a rarely reported manifestation of metastatic disease. Neoplasms were high grade carcinomas. Tumor emboli were present within portal branches ranging 0.12-2.9 mm in diameter and were free floating or attached to the vascular wall, with or without varying degrees of superimposed organization. In one case, intravascular tumor necrosis was prominent and appeared as granular casts with superimposed dystrophic calcification and/or entrapped foamy histiocytes. There were associated geographical areas of parenchymal (4 cases) and tumor (1 case) ischemic necrosis with a multifocal and regional topographic distribution. An associated predominant pattern of intrasinusoidal tumor infiltration (with or without fibrosis) was present in 3 cases, whereas the fourth case had underlying micronodular cirrhosis, providing ancillary evidence for preexisting altered intrahepatic microcirculation. The literature on fatal hepatic failure resulting from neoplasia is reviewed with a reassessment of its pathobiological significance. PMID- 11432672 TI - Muscle plasma membrane changes in dystrophin gene exon 52 knockout mouse. AB - Changes in muscle plasma membranes in mice lacking exon 52 of the dystrophin gene (mdx52 mouse) were studied using the freeze-fracture technique. The extensor digitorum longus (EDL) muscle plasma membrane of the mdx52 mouse at 8 weeks of age showed significantly increased caveola density (p < 0.05 by two-tailed t test) and significantly decreased densities of intramembranous particles (IMPs), orthogonal arrays (OAs) and orthogonal array subunit particles (OASPs) (p < 0.05 by two-tailed t-test, p < 0.01 by Wilcoxon rank-sum test, p < 0.05 by two-tailed t-test, respectively) on the protoplasmic face when compared with those of control EDL muscles. These changes are more similar to those seen in DMD than those in the mdx mouse at the same age as reported previously. Thus, the gene abnormality in the different exon of the mouse dystrophin gene seems to induce somewhat different changes in the muscle plasma membrane. PMID- 11432673 TI - Primary adenocarcinoma of the arytenoid: an ultrastructural study. AB - Primary adenocarcinoma of the larynx is a rare neoplasm that tends to spread to both regional lymph nodes and distant sites. A case of primary adenocarcinoma of the arytenoid in a 74-year-old man is presented. The tumor was evaluated by light and electron microscopy. A high percentage of intranuclear pseudoinclusions (more than 20% of the neoplastic cells) was a peculiar characteristic of the tumor. To the best of our knowledge, such a feature has not been reported previously and should be considered a hallmark of more aggressive behavior. PMID- 11432674 TI - Vitamin A deficiency among children--Federated States of Micronesia, 2000. AB - Vitamin A, a fat-soluble, heat-stable nutrient (retinol) derived from animal sources and certain fruits and vegetables, forms the basic component of retinal pigments and plays a vital role in optimal health, growth, and development. Vitamin A deficiency (VAD) (serum retinol < or = 20 microg/dL [< or = 0.7 micromol/L] for subclinical VAD) can substantially increase the risk for childhood mortality from infectious and noninfectious causes. VAD impairs the mobilization and transport of iron and is usually associated with anemia and reduced growth. VAD is a major public health problem in parts of Africa, Asia, Latin America, and the Western Pacific. In Chuuk and Pohnpei, two of the four Federated States of Micronesia (FSM) (2000 population: 107,008), nutrition surveys during the early 1990s documented VAD prevalences among the highest in the world (CDC, unpublished data, 1991; U.S. Public Health Service, unpublished data, 1994). In response to these findings, FSM health authorities, with support of the United Nations Children's Fund (UNICEF), began distributing vitamin A supplements in 1993 and 1998 in Chuuk and Pohnpei, respectively. In November 1999, FSM requested assistance from CDC in VAD assessment surveys of children in Kosrae and Yap, the other two FSM states. This report summarizes levels of serum retinol and prevalence of VAD and other indicators of nutritional status among children aged 24-59 months in Kosrae and Yap. The findings indicated low serum retinol levels and high VAD prevalences but no substantial stunting or wasting. A comprehensive, long-term national strategy is needed in FSM to promote sustained improvement in vitamin A status. PMID- 11432675 TI - Measles incidence before and after supplementary vaccination activities--Lusaka, Zambia, 1996-2000. AB - Zambia is a sub-Saharan African country (2000 population: nine million) with approximately 10% of the population residing in the capital of Lusaka. In Zambia, measles is one of the five major causes of morbidity and mortality among children aged <5 years. During 1991-1999, the annual number of reported measles cases ranged from 1698 to 23,518. In August 1999, supplementary vaccination activities (SVAs) were conducted in Lusaka among children aged 9 months-4 years. This report summarizes measles incidence, measured by the number of patients presenting to selected medical facilities, before and after SVAs and suggests that substantial measles transmission continued despite this intervention. To improve measles control in Zambia, nationwide supplementary measles vaccination is planned for children aged 9 months-14 years in 2002. PMID- 11432676 TI - Antimicrobial prophylaxis with ceftriaxone for prevention of early postoperative infections after 49 liver transplantations. AB - Infection remains a major problem for individuals who undergo solid organ transplantation and liver transplant recipients are particularly susceptible to infectious complications with a high morbidity and mortality rate. The risk of these infections is determined by previous or future environmental exposures as well as the patient's immune status. We report here the results of an open prospective study involving 49 consecutive liver transplantations, undertaken to evaluate the efficacy of ceftriaxone in the prevention of early postoperative infectious complications. Antimicrobial prophylaxis was done using a single dose of ceftriaxone (2 g i.v.) given at the induction of anesthesia and then 2 further once-daily doses were administered 2 days postoperatively. Early postoperative bacterial infection rate was 43.5% (20/46); this result is comparable or even lower than those documented in other studies. This study, even though open and non-comparative, showed that a once-daily regimen containing ceftriaxone provides adequate antimicrobial prophylaxis and significant cost-savings in comparison with multiple-dose prophylactic regimens in patients undergoing liver transplantation. PMID- 11432677 TI - Antimicrobial prophylaxis with ceftriaxone in laparoscopic cholecystectomy: a 7 year clinical experience involving 3,603 patients. AB - This study is one of the first in the literature with the aim of assessing the effectiveness of antimicrobial prophylaxis with a single preoperative dose of long-acting ceftriaxone in a retrospective analysis of 3,603 patients undergoing laparoscopic cholecystectomy. All patients who underwent laparoscopic cholecystectomy between October 1990 and December 1997 were reviewed. Antimicrobial prophylaxis with a single dose of ceftriaxone (1 g) was given intravenously at the induction of anesthesia. Patients were closely monitored for infections until 4 weeks after surgery. Postoperative infections occurred in 44 of the 3,603 patients undergoing laparoscopic cholecystectomy, with an infection rate of 1.22%. The infectious complications mainly comprised wound infections (n=13) but also included intra-abdominal abscesses (n=3), pneumonia (n=9), urinary tract infections (n=3) and other infections (n=16). The results of this retrospective and non-randomized study, show that a single-shot regimen containing ceftriaxone may be a cost-effective measure in preventing postoperative infections in patients undergoing laparoscopic cholecystectomy. PMID- 11432678 TI - Ceftriaxone for surgical prophylaxis: clinical experience and pharmacoeconomics. PMID- 11432679 TI - Short-Term prophylaxis with ceftriaxone plus metronidazole in esophageal cancer surgery. AB - An open prospective study was carried out in 82 consecutive patients undergoing resective surgery for esophageal cancer from January 1995 to July 1996. Antimicrobial prophylaxis was done using a single dose of ceftriaxone (2 g i.v.) given at the induction of anesthesia in combination with metronidazole (0.5 g i.v.). Two further doses of metronidazole were administered 8 and 16 hours postoperatively. Fourteen patients (17%) experienced postoperative infections. This study, even though open and non-comparative, confirms that ceftriaxone given as a single-dose plus metronidazole provides adequate prophylaxis and significant cost-savings in comparison with multiple-dose prophylactic regimens in patients undergoing major surgery for esophageal cancer. Furthermore, the single-dose regimen reduces the workload for the nursing staff, the risk of side effects, and the possibility of selecting resistant strains. PMID- 11432680 TI - Ceftriaxone as short-term antibiotic prophylaxis in orthopedic surgery: a cost benefit analysis involving 808 patients. AB - This paper reports the results observed in 477 consecutive patients (from June to December 1996) undergoing orthopedic surgery and treated with a single preoperative dose of ceftriaxone, a long-acting cephalosporin, in comparison with those observed in a control group of 331 patients (from January to December 1995) receiving no antibiotic prophylaxis. Patients receiving ceftriaxone did better than those receiving no antibiotic prophylaxis in terms of postoperative infection rate (1.3% vs. 11.8%) and duration of hospital stay (on average 3.25 vs. 7.09 days, respectively). More importantly, administering ceftriaxone as a single preoperative injection led to a dramatic savings in terms of nursing effort, time and cost of prophylaxis per patient. It is concluded that the clinical results and economic implications seem to justify the use of ceftriaxone as a routine prophylactic antibiotic for patients undergoing orthopedic surgery. PMID- 11432681 TI - A retrospective study on the efficacy of short-term perioperative prophylaxis in abdominal surgery for hernia repair in 1,254 patients. AB - Between 1992 and 1999 1,254 patients requiring hernioplasty came to our Department of Surgery, at San Giovanni Valdarno Hospital, Arezzo. In 94% of cases a prosthetic device was inserted. All patients received a preoperative antibiotic prophylaxis with ceftriaxone, administered intravenously in a single 2 g dose, before surgery. We have retrospectively evaluated the incidence of postoperative infections in these patients: no surgical wound infection occurred, while we observed 10 cases of systemic infections, 5 respiratory tract infections and 5 urinary tract infections, all of which were successfully treated with antibiotics. Tolerability was good: no adverse reaction to ceftriaxone was observed. In this retrospective study ceftriaxone was shown to be effective in preventing the onset of postoperative infections. PMID- 11432682 TI - Comparative study of ceftriaxone versus cefazolin plus clindamycin as antibiotic prophylaxis in elective colorectal surgery. AB - A comparative study of 103 consecutive patients who required antibiotic prophylaxis for elective colorectal surgery was carried out. All eligible patients received either ceftriaxone (2 g) as a single intravenous dose at anesthetic induction or cefazolin (1 g) plus clindamycin (0.6g) administered intravenously at anesthetic induction and for two more doses at 8-hourly intervals for a total of 3 days. The incidence of postoperative wound infections was 6.9% in the ceftriaxone group and 11.1% in the cefazolin plus clindamycin group. Single-dose ceftriaxone proved to be a safe and cost-effective form of antibiotic prophylaxis for elective colorectal surgery. PMID- 11432683 TI - Effects of cocaine on extracellular dopamine and serotonin levels in the nucleus accumbens. AB - RATIONALE: Although the rewarding effects of cocaine are generally attributed to its ability to increase dopamine (DA) transmission, cocaine demonstrates approximately equal affinity for dopamine and serotonin (5-HT) transporters in vitro. However, there have been few direct systematic comparisons of the effects of cocaine on DA and 5-HT transmission in vivo. OBJECTIVES: The present experiments compared the effects of systemic cocaine administration, local cocaine infusion and the systemic administration of cocaine during infusion on extracellular levels of DA and 5-HT in the nucleus accumbens (NAc). METHODS: In vivo microdialysis in awake unrestrained rats was used to measure the effects of systemic administration and local infusion of cocaine on extracellular DA and 5 HT levels simultaneously in the NAc. RESULTS: Systemic cocaine (10-25 mg/kg, IP) dose-dependently increased DA and 5-HT levels, but the increase in DA was larger than for 5-HT at 18 mg/kg. Infusion of cocaine (0.1-10.0 mM) into the NAc increased both DA and 5-HT levels, but the effect on DA was larger than 5-HT at 0.1 and 3 mM cocaine. The influence of cocaine on DA and 5-HT somatodendritic autoreceptors was examined when cocaine (25 mg/kg) was administered systemically during cocaine infusion. The increase in DA and 5-HT levels during cocaine infusion was attenuated by the systemic injection of cocaine during cocaine infusion, but the decrease of 5-HT was greater than that for DA. CONCLUSIONS: Cocaine produced a larger impact on DA than 5-HT neurotransmission under specific conditions. A series of physiological mechanisms, i.e. terminal density, neurotransmitter interactions and somatodendritic regulation, are discussed as factors responsible for facilitating cocaine's effects on DA relative to 5-HT. PMID- 11432684 TI - Dropout rates in randomised antipsychotic drug trials. AB - RATIONALE: It has been assumed that new atypical drugs improve treatment compliance due to fewer adverse effects. Data supporting this assumption are scarce. OBJECTIVES: The aim of this study was to study attrition rates in randomised controlled trials of oral administration of conventional antipsychotic drugs, atypical antipsychotic drugs and placebo. METHODS: The database of the Schizophrenia Module of the Cochrane Library was utilised for the present study. The data in the Cochrane Module are collected by identifying relevant randomised controlled trials from several electronic databases and other sources. Number of dropouts was defined as patients leaving the study preterm due to any reason. RESULTS: Data from 328 treatment groups, consisting of 18,585 randomised subjects from 163 drug trials, were entered in the analysis. One-third of the subjects had dropped out of the trials. The dropout rates significantly increased for each calendar year. Year of trial publication, type of drug and trial length remained statistically significant contributors to dropout rates. In a model incorporating year of publication and trial length, placebo groups and groups treated with conventional antipsychotics had significantly higher attrition rates than groups treated with atypical drugs. When clozapine-treated groups were excluded from the analysis, no statistically significant advantage for atypical drugs over conventional drugs remained. CONCLUSIONS: Trial data implicate that a better compliance can be achieved by favouring atypical drugs rather than conventional alternatives in the treatment of schizophrenia. However, this effect is found only when groups treated with the atypical antipsychotic clozapine are included in the analysis. Our study did not find evidence for a statistically significant superiority in acceptability of novel atypical drugs when compared to conventional antipsychotics. PMID- 11432685 TI - Effect of aerobic exercise on behavioral and neuroendocrine responses to meta chlorophenylpiperazine and to ipsapirone in untrained healthy subjects. AB - RATIONALE: Several clinical studies suggest antidepressive and anxiolytic effects of regular aerobic exercise. OBJECTIVES: The present study examines the effects of a 10-week protocol of moderate aerobic exercise (3-4 miles jogging 3 times per week) on central serotonergic receptor sensitivity. METHODS: Neuroendocrine challenges using oral doses of meta-chlorophenylpiperazine (m-CPP; 0.4 mg/kg), ipsapirone (0.3 mg/kg), and placebo were performed in 12 untrained healthy volunteers before and after 10 weeks of moderate aerobic exercise. RESULTS: Before training, administration of the non-selective serotonergic agonist m-CPP, which exerts a number of well-reproducible effects mainly via its action on 5 HT2C receptors, was associated with a significant increase of cortisol and prolactin (but not adrenaline or noradrenaline) in comparison with the placebo condition. After the 10-week training period, administration of m-CPP was followed by a blunted cortisol response which was not significantly increased in comparison to the placebo challenge. In contrast, the increases of cortisol observed after administration of the 5-HT1A agonist ipsapirone were of the same magnitude during the pre- and post-training challenge sessions. The behavioral response to ipsapirone and the mean maximal increases of plasma adrenaline and noradrenaline did not change during the training period. CONCLUSIONS: Regular aerobic exercise is associated with a blunted cortisol response to m-CPP, which might reflect a downregulation of central 5-HT2C receptors. PMID- 11432686 TI - Anxiogenic-like effects of opiate withdrawal seen in the fear-potentiated startle test, an interdisciplinary probe for drug-related motivational states. AB - RATIONALE: Anxiety-like effects may be universal to withdrawal from drugs of abuse. The study of withdrawal would benefit from the acoustic startle response (ASR), a discrete, cross-species reflex which is increased by fear-related states. However, existing reports of opiate-related effects on baseline ASR have not validated ASR as a measure of drug-related motivation. OBJECTIVE: The effects of opiate treatment and withdrawal were examined using fear-potentiated startle, a startle test more sensitive to fear than baseline changes. METHODS: Fear conditioned rats were treated with Alzet osmotic pumps delivering 0.25 mg/kg per day fentanyl or placebo pumps. Experiment I examined changes before and during opiate treatment on locomotor activity and baseline, prepulse inhibition, and fear-potentiated startle. Experiment 2 examined the same responses during withdrawal precipitated after 4-7 days of treatment using IV naloxone. RESULTS: Experiment 1 revealed an attenuated fear-potentiated startle on the first test after the start of fentanyl treatment (4 h); this was not seen on subsequent tests and suggested tolerance to this acute effect. Experiment 2 found an enhancement of fear-potentiated startle precipitated in fentanyl-treated rats after injection of 0.025 and 0.16 mg/kg naloxone; this was not seen at 1 mg/kg naloxone, even though more physical withdrawal signs were most prevalent at this dose. In neither experiment did locomotor activity, baseline ASR, or prepulse inhibition of the ASR show any treatment effect. CONCLUSIONS: Fear-potentiated startle may provide a specific and valid measure of anxiety-like effects of drug withdrawal. Discussed were conditions needed to see this effect and the relevance of the findings for different mechanisms of withdrawal discomfort. PMID- 11432687 TI - Effects of central noradrenaline depletion by the selective neurotoxin DSP-4 on the behaviour of the isolated rat in the elevated plus maze and water maze. AB - RATIONALE: Social isolation of the rat from weaning influences behaviour following central noradrenaline (NA) depletion by the selective neurotoxin N-(2 chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4). OBJECTIVES: The study characterised the effects of DSP-4 on the behaviour of isolates in the elevated plus maze and water maze. METHODS: Male Lister hooded rats were reared singly or in groups after weaning. Two weeks postweaning, the rats were injected with DSP-4 (25 mg/kg, i.p.) or saline. From week 4, rats were tested in the plus maze and in the water maze. RESULTS: DSP-4 significantly reduced cortical and hippocampal NA but had no effect on hypothalamic NA. Isolation rearing alone had no significant effects on behaviour in the elevated plus maze but enhanced retention of platform placement in the water maze as measured by increased entries to the platform annulus during the probe test. DSP-4 in group-reared rats increased activity in the open arms and increased general activity in the elevated plus maze with no effect on water maze performance. DSP-4-treated isolates spent less time in the open arms and were hypoactive in the plus maze compared to group-reared DSP-4 treated rats, and had impaired retention of spatial memory in the water maze compared to isolate controls. CONCLUSIONS: DSP-4 treatment had an 'anxiolytic' effect in group-reared rats in the elevated plus maze. In the water maze, isolation rearing enhanced retention of spatial information, an effect normalised by NA depletion. The results demonstrate the importance of noradrenergic function in the regulation of responsiveness to environmental cues. PMID- 11432688 TI - Long-term effects of clomipramine and fluoxetine on dorsal periaqueductal grey evoked innate defensive behaviours of the rat. AB - RATIONALE: The defensive responses induced by electrical stimulation of dorsal periaqueductal grey (DPAG) of the rat have been proposed as a model of panic attacks in humans. OBJECTIVE: The present experiments were carried out to evaluate the long-term effects of clinically effective panicolytics on these responses. METHODS: Rats that had electrodes implanted into the DPAG were treated for 21 days with clomipramine (CLM; 5, 10 and 20 mg/kg), fluoxetine (FLX; 1 and 5 mg/kg) or saline. Long-term effects were assessed prior to the treatment to avoid acute drug effects. Threshold logistic functions of defensive responses were compared by likelihood ratio coincidence tests. RESULTS: CLM attenuated DPAG evoked defensive behaviours at a time-course similar to that observed in panic therapy. Administration of 10 mg/kg CLM for 21 days produced significant increases in the thresholds of immobility (24%), running (49%) and jumping (45%). Splitting of running into single responses disclosed selective threshold increases of galloping (75%) and trotting (138%) with 5 and 10 mg/kg, respectively. Thresholds of micturition were markedly increased (87%) by 5 mg/kg CLM. In turn, FLX (1 mg/kg) virtually abolished the galloping response. No threshold changes were observed following the long-term administration of the higher dose of either CLM or FLX. Saline-treated rats had a significant increase (35%) in galloping thresholds only. CONCLUSIONS: The present data partially validate the DPAG-evoked defence reaction of the rat as a model of panic attacks in humans. Attenuation of galloping by lower doses of FLX and CLM also suggests the prominent modulation of this response by serotonin. PMID- 11432689 TI - Spontaneous activity as a contingency-controlled behavior within an operant context: alprazolam concentration-effect relations after subcutaneous administration in rats. AB - RATIONALE: Environmental factors affect serum drug concentration-effect relations. For example, after midazolam administration, longer pre-session delays imposed in experimental chambers produced differential concentration-effect relations compared to those of shorter delays. OBJECTIVES: To evaluate the extent to which serum concentrations determine alprazolam's effects on spontaneous activity in the presence and absence of a differential reinforcement of low rate (DRL 45-s) contingency using pharmacokinetic-pharmacodynamic analysis. Serum concentrations reported here were simulated from our published pharmacokinetic parameters for alprazolam. METHODS: One group (n=8) was used to investigate alprazolam's effects on spontaneous activity within the DRL contingency by placing an activity platform beneath each operant chamber to monitor concurrently both spontaneous activity (large and small movements) and DRL performance (shorter-response and reinforcement rates) in 3-h sessions; a parallel group (n=7) was used without the operant context. The concentration-effect relation of the reinforcement rate was compared and contrasted with those of spontaneous activity. RESULTS: Alprazolam decreased large and small movements within the DRL contingency, which corresponded to that of reinforcement rates under the DRL 45-s schedule. In contrast, without the DRL contingency, alprazolam's effects on small movements were short-lived (i.e., 30 min) and no effects on large movements were detected. Hence, the predicted concentration-effect relations for the reinforcement rate function described those of spontaneous activity well within the operant context, but not those without the operant context. Furthermore, the latter showed no correlation between serum alprazolam concentration and large movements; a significant, but low negative correlation for small movements was observed. CONCLUSIONS: The duration of alprazolam's action was dependent on not only dose size but also the behavioral measure examined. By imposing the DRL contingency, spontaneous activity behaves as an ideal pharmacodynamic measure (i.e., continuous, sensitive, and objective). PMID- 11432690 TI - Environmental enrichment decreases intravenous self-administration of amphetamine in female and male rats. AB - RATIONALE: Previous work has shown that environmental enrichment alters amphetamine-induced locomotor activity and conditioned place preference. OBJECTIVE: The present study examined the effect of environmental enrichment on amphetamine self-administration. METHODS: Female and male rats were raised from 21 days of age in one of three different conditions: an enriched condition (EC) containing novel objects and social partners, a social condition (SC) containing social partners only, or an isolated conditioned (IC) without objects or social partners. Beginning at 51 days of age, rats were then tested for operant responding for a sucrose reinforcer using an incremental fixed ratio (FR) requirement across four sessions. Rats were then implanted with a chronic indwelling intravenous catheter and were allowed to self-administer amphetamine (0.03 or 0.1 mg/kg per infusion) for five FR1 sessions, followed by a progressive ratio (PR) session. RESULTS: EC rats initially showed an increase in sucrose reinforced responding relative to IC rats and this environment-induced difference was greater in females than in males. However, in both sexes, the environment induced difference in sucrose-reinforced responding dissipated completely across repeated sessions. With amphetamine self-administration, both EC and SC rats earned fewer infusions than IC rats across repeated FRI sessions using the low dose of amphetamine (0.03 mg/kg per infusion), but not using the higher dose of amphetamine (0.1 mg/kg per infusion). EC rats also earned fewer self-infusions of the low amphetamine dose on the PR session relative to IC rats. The effects of environmental enrichment on amphetamine self-administration were similar in both females and males. CONCLUSION: These results suggest that environmental enrichment may serve as a protective factor for reducing amphetamine self administration. PMID- 11432691 TI - Behavioral characterization of Co 134444 (3alpha-hydroxy-21-(1'-imidazolyl)-3beta methoxymethyl-5alpha- pregnan-20-one), a novel sedative-hypnotic neuroactive steroid. AB - RATIONALE: Neuroactive steroids have been shown to exhibit a wide range of behavioral activities that are similar but not identical to those of benzodiazepines. These activities include anticonvulsant, anxiolytic and sedative hypnotic effects. OBJECTIVE: The purpose of the present study was to characterize Co 134444 (3alpha-hydroxy-21-(1'-imidazolyl)-3 -methoxymethyl-5alpha-pregnan-20 one), a novel sedative-hypnotic neuroactive steroid, in a variety of behavioral procedures. METHODS: Anticonvulsant effects were determined by the ability to protect against pentylenetetrazol- and maximal electroshock-induced seizures in mice and rats. Anxiolytic-like effects were determined using a punished drinking procedure in rats. Ataxic effects were determined using a horizontal wire procedure in mice and a rotorod procedure in mice and rats. The discriminative stimulus effects were evaluated in rats trained to discriminate pregnanolone from vehicle. RESULTS: Co 134444 exhibited oral anticonvulsant activity against pentylenetetrazol and maximal electroshock with ED50s of 9.8 and 20.6 mg/kg, respectively, in mice and 23.6 and 25.3 mg/kg, respectively, in rats. Anxiolytic like efficacy was observed at a dose as low as 3.0 mg/kg, PO, in rats. Ataxic effects were observed with rapid onset and short duration. TD50s were 17.4 and 21.2 mg/kg orally in mice in the horizontal wire and rotorod procedures, respectively, and 39.0 mg/kg in rats using the rotorod. Co 134444 completely substituted for pregnanolone as a discriminative stimulus with little effect on response rate. CONCLUSIONS: Co 134444 exhibits a wide variety of behavioral effects; however, its rapid onset and short duration are consistent with its potential use as a sedative-hypnotic drug. PMID- 11432692 TI - Compounds with affinity for serotonergic receptors in the treatment of premenstrual dysphoria: a comparison of buspirone, nefazodone and placebo. AB - RATIONALE: It is well established that serotonin reuptake inhibitors (SRIs) are effective for the treatment of premenstrual dysphoria (PMD), but the receptor subtype(s) mediating this effect of serotonin have yet not been identified. OBJECTIVE: In this trial, the possible efficacy of buspirone, a partial 5HT1A receptor agonist, and nefazodone, a combined SRI and 5HT2 receptor antagonist, was evaluated in women with PMD. METHODS: After a three-menstrual-cycle screening phase, patients were randomised to buspirone (n=19), nefazodone (n=22) or placebo (n=22). During the first two treatment cycles, patients were taking the drug during the luteal phase only (mean +/- SD daily dose of buspirone: 21 +/- 6 mg; nefazodone: 228 +/- 54 mg). During the subsequent two cycles, the medication was taken each day of the menstrual cycle (mean daily dose of buspirone: 27 +/- 10 mg; nefazodone: 304 +/- 95 mg). RESULTS: With respect to self-rated global improvement, buspirone (P<0.001) but not nefazodone was significantly superior to placebo. While buspirone appeared to reduce self-rated irritability (visual analogue scale) more effectively than placebo, other self-rated symptoms did not differ markedly between the groups. The side-effects were mild, and sexual dysfunction was not significantly more common in patients given buspirone or nefazodone than in those given placebo. CONCLUSION: It is suggested that buspirone is mildly effective for premenstrual irritability. In patients experiencing sexual dysfunction when treated with an SRI, buspirone may be a useful alternative. PMID- 11432693 TI - Lack of phencyclidine-induced effects in mice with reduced neuronal nitric oxide synthase. AB - RATIONALE: Phencyclidine (PCP) is widely used as an animal model of schizophrenia, because in humans it can induce positive and negative symptoms associated with schizophrenia. PCP is an antagonist of N-methyl-D-aspartate receptors, which are associated with the nitric oxide (NO) system. OBJECTIVE AND METHODS: The primary objective was to determine whether neuronal NO synthase (nNOS) is involved in PCP-induced behaviours and neuronal activation, as measured by the expression of c-Fos. After characterizing a PCP mouse model (dose-response study, Experiment 1), we measured PCP-induced effects in mice treated with nNOS antisense oligodeoxynucleotides (AS-ODNs) (Experiment 2), and in nNOS knockout (nNOS-/-) mice (Experiment 3). RESULTS: PCP 5 mg/kg induced the maximum behavioural effects of all doses tested, consisting of hyperlocomotion, stereotyped turning behaviour, without the presence of ataxia. PCP also induced an increase in Fos-like immunoreactivity (Fos-LIR) in the frontal cortex, as well as in the midline limbic (thalamic and hypothalamic nuclei) areas. In the AS-ODN treated mice, PCP induced less behaviour when compared to water-treated controls. In the nNOS-/- mice, PCP induced less behaviour and a decrease in Fos-LIR in the frontal cortex and midline limbic areas, when compared to wild-type littermate controls. CONCLUSIONS: Our findings suggest that the frontal cortex and midline thalamic brain regions are involved in PCP-induced effects in mice. Furthermore, we show that an intact nNOS system is necessary to obtain PCP-induced effects. This may implicate nNOS as a viable drug target in the treatment of schizophrenia. PMID- 11432694 TI - Reduced serotonin transporter binding in binge eating women. AB - RATIONALE: There is evidence that abnormalities in brain dopamine, norepinephrine and serotonin metabolism may play an important role in binge eating. Serotonin active antidepressant drugs have also been found to decrease binge eating. OBJECTIVE: We investigated serotonin transporter binding in obese binge-eating women. Eleven obese binge-eating and seven obese control women participated in the study. The subjects were not taking any medication known to affect serotonin (5-HT) transporters. METHODS: We used single-photon emission tomography (SPECT) with the radioligand 123I-labelled nor-beta-CIT, which specifically labels 5-HT transporters. RESULTS: Obese binge-eating women showed significantly decreased 5 HT transporter binding in the mid-brain compared with obese controls (2.1 +/- 0.5 versus 2.9 +/- 0.5, respectively). CONCLUSIONS: SPECT imaging with a ligand specific for 5-HT transporters can be used to assess altered serotonin transporter binding in the living human brain. The results tentatively suggest that 5-HT transporter binding is decreased in binge-eating women. PMID- 11432695 TI - Sensitivity to the effects of pharmacologically selective antidepressants in different strains of mice. AB - RATIONALE: Recent advances in neurobehavioral genetics have increased the importance of research on the behavioral patterns of different mouse strains. A comprehensive comparison of inbred and outbred mouse strains was conducted to provide information on the range of performance and pharmacological effects in the forced swimming test, a behavioral test commonly used to measure the effects of antidepressant drugs. OBJECTIVES: Baseline performance and pharmacological responses to desipramine, a selective norepinephrine reuptake inhibitor, and fluoxetine, a selective serotonin reuptake inhibitor, were compared in seven inbred and four outbred mouse strains in the forced swimming test. METHODS: Swim sessions were conducted by placing mice in individual glass cylinders filled with water for 6 min. The duration of behavioral immobility during the last 4 min of the test was scored from videotapes. RESULTS: A 10-fold range of immobility values and coefficient of variation supported the existence of substantial behavioral differences between mouse strains in baseline performance in the FST. In general, inbred strains demonstrated lower variability than outbred strains. Desipramine dose-dependently reduced immobility in seven of the 11 strains tested, with DBA/2J and the C57BL/6J mice showing greater sensitivity than the other strains. In contrast, fluoxetine reduced immobility in only three out of the 11 strains tested, DBA/2J, BALB/cJ and NIH Swiss mice. CONCLUSIONS: Background strain is a critical variable in determining baseline performance and the sensitivity to different types of antidepressant drugs in the mouse FST. The use of such mouse strains may provide information on the genetic basis for strain differences in depressive behavior and differential sensitivity to diverse classes of antidepressants. PMID- 11432696 TI - Low-dose citalopram as a 5-HT neuroendocrine probe. AB - RATIONALE: Intravenous administration of the selective serotonin re-uptake inhibitor, citalopram (20 mg), is known to increase plasma prolactin (PRL) and cortisol in human subjects. This suggests that citalopram may be a useful tool to probe brain serotonin function. OBJECTIVE: To find out whether lower doses of intravenous citalopram would be sufficient to increase plasma prolactin and cortisol. METHODS: Eleven subjects were tested on three occasions in a double blind, cross-over design receiving: (a) placebo, (b) citalopram 5 mg and (c) citalopram 10 mg infused intravenously over a 30-min period. A further six subjects received intravenous citalopram (10 mg) on two occasions receiving in addition the 5-HT2A2C receptor antagonist, cyproheptadine (4 mg orally) or placebo, 6 h before each infusion in a double-blind, randomised, cross-over design. Plasma PRL and cortisol levels were measured before and for 150 min after the infusion. RESULTS: Citalopram increased plasma PRL and cortisol in a dose related manner. Cyproheptadine lowered baseline PRL and cortisol but did not attenuate the endocrine responses to citalopram. Citalopram infusions were well tolerated. CONCLUSIONS: Low-dose citalopram has potential utility as a neuroendocrine challenge test. The endocrine responses to citalopram are probably not mediated predominantly by 5-HT2A/2C receptors. PMID- 11432697 TI - Geochemical and cosmochemical materials. PMID- 11432698 TI - Coatings. PMID- 11432699 TI - Process analytical chemistry. PMID- 11432700 TI - Water analysis. PMID- 11432701 TI - Forensic science. PMID- 11432702 TI - Biological monitoring: exquisite research probes, risk assessment, and routine exposure measurement. PMID- 11432704 TI - Petroleum and coal. PMID- 11432703 TI - Environmental analysis. PMID- 11432705 TI - Pharmaceuticals and related drugs. PMID- 11432706 TI - Hair dyes and bladder cancer: an update. PMID- 11432707 TI - The gastro-oesophageal reflux disease complex in sub-Saharan Africa. AB - Epidemiological and clinical studies that have reported on gastro-oesophageal reflux disease (GERD), Barrett's oesophagus, oesophageal adenocarcinoma and Helicobacter pylori infection in sub-Saharan Africa were reviewed. The data indicate that Barrett's oesophagus is rare and oesophageal adenocarcinoma uncommon in all regions of sub-Saharan Africa studied (South Africa, Ethiopia, Nigeria, Zimbabwe, Kenya and Uganda). Hiatus hernia is also uncommon. There are too few reports of GERD to allow comment. The overwhelming majority of oesophageal cancers are squamous cell type. H. pylori infection is ubiquitous with an overall prevalence of 61-100%. It is concluded that although urbanization has resulted in an increase of risk factors associated with GERD, which would be expected to lead to an increase in this disease among Africans, this increase has not happened. It is believed that the critical factor preventing GERD in black Africans is H. pylori infection, which is usually acquired in childhood, is lifelong and is probably protective for the oesophagus. PMID- 11432708 TI - Molecular genetic abnormalities in premalignant lung lesions: biological and clinical implications. AB - Lung cancer is a leading cause of cancer death worldwide; however, despite major advances in cancer treatment during the past two decades, the prognostic outcome of lung cancer patients has improved only minimally. This is largely due to the inadequacy of the traditional screening approach, which detects only well established overt cancers and fails to identify precursor lesions in premalignant conditions of the bronchial tree. In recent years this situation has fundamentally changed with the identification of molecular abnormalities characteristic of premalignant changes; these concern tumour suppressor genes, loss of heterozygosity at crucial sites and activation of oncogenes. After considering the morphological modifications that occur in premalignant lesions of the bronchial tree, we analyse the alterations occurring in a series of relevant genes: p53 and its functional regulation by MDM2 and p14ARF, p16INK4, p15INK4b, FHIT, as well as LOH at important sites such as 3p, 8p, 9p and 5q. Activation of oncogenes is considered for K-ras, the cyclin D1, the heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNPA2/B1), and finally the c-myc oncogene. The expression of c-myc is influenced strongly by the presence of growth factors (GFs), among which EGF is of prime importance, as well as its receptor coded for by the c-erbB-2 oncogene. Basic knowledge at the molecular level has extremely important clinical implications with regard to early diagnosis, risk assessment and prevention, and therapeutic targets. The novel techniques for early diagnosis and screening of premalignant lung lesions, such as fluorescence bronchoscopy, endobronchial ultrasound, spiral computed tomography combined with precise spatial localization techniques, should basically change the approach to the problems raised by this disease and allow for an increased discovery rate of incipient lesions. Sequential applications will lead to the identification of individuals/populations at high risk, while the availability of accurate 'intermediate end points' will enable the effects of preventive trials to be monitored. Finally, the same molecular abnormalities may serve as 'starting points' for innovative treatments designed to restore the altered functions to normality. Recent developments in our knowledge and understanding of the molecular genetic abnormalities in premalignant lung lesions open an era of hope. PMID- 11432709 TI - Gender-specific incidence trends in lung cancer by histological type in Sweden, 1958-1996. AB - We used the Swedish Family-Cancer Database to examine the time trends of lung cancer in Sweden by histological type, with special reference to gender. A total of 45,297 lung cancer cases were analysed. The overall age-adjusted incidence rates of squamous cell carcinoma (SCC) in men peaked in the 1980s and then slightly decreased, while corresponding rates in women increased steadily. The incidence of adenocarcinoma increased in both genders. The male-to-female incidence ratio for SCC was 12.4 in the 1960s and 3.6 in the 1990s. For adenocarcinoma the ratio was close to 1.5 throughout the period. Regression analysis indicated that the birth cohort of the 1940s was at the highest risk for adenocarcinoma in men and for all types of lung cancer in women. Lung cancer in parents was a risk factor for offspring. In conclusion, the data, particularly on women, suggest that modern cigarettes induce lung adenocarcinoma and SCC in a proportion of 1:0.6. This proportion was 1:3.7 among men diagnosed in the 1960s. The incidence ratio of 1.3 for adenocarcinoma between men and women in the 1990s is consistent with the smoking prevalence data a few decades ago, suggesting equal sensitivity of both genders to tobacco-induced lung cancer. PMID- 11432710 TI - Phytosterols have an unfavourable effect on bacterial activity and no evident protective effect on colon carcinogenesis. AB - The effects of physiological dietary phytosterol supplements on intestinal microflora activity and faecal sterols and their capacity to protect rats fed a normal or high saturated fatty-acid diet against tumour development were studied. A group of 80 female Wistar rats were fed an 8% lipid diet for 4 weeks (adaptation period) and then randomly assigned in a factorial experimental design study to diets containing 8% or 24% hydrogenated coconut oil, with or without a 24-mg/day/rat phytosterol supplement. They were instilled intrarectally with saline or methyl-nitroso-urea (MNU). Faecal sterol output was analysed for one week each month. Pathological analysis was done at the end of the 30-week experiment. Animals treated with MNU and given phytosterol supplements had tumour frequencies (8/20) similar to those not fed phytosterols (11/20). The fat supplemented diet had no significant influence. Colonic glands were found in area of lymphoid follicles in all the groups, but were more frequent in rats on high fat diets (P < 0.01). The coprostanol and the cholesterol excretion of the phytosterol-supplemented rats was significantly enhanced. Therefore phytosterols have an unfavourable effect on bacterial activity. These data confirm the capacity of phytosterols to decrease cholesterol absorption, but indicate that a large excess of phytosterol must be avoided until further research on its effects on carcinogenesis has been done. PMID- 11432711 TI - Measurement variability of plasma beta-sitosterol and campesterol, two new biomarkers for cancer prevention. AB - Phytosterols are plant sterols that are structurally similar to cholesterol and are characterized by anti-carcinogenic and anti-atherogenic properties. Beta sitosterol and campesterol are the predominant phytosterols in blood. The present study aimed to analyse the reproducibility and overtime reliability of plasma beta-sitosterol and campesterol measurements. In order to study the reproducibility of the measurement (technical variability), three healthy premenopausal women donated a sample of their blood. Each blood sample was subdivided into six aliquots and analysed within the same run by the same laboratory technician. The intraclass correlation coefficients (ICCs) of the assay for plasma beta-sitosterol and campesterol were 0.88 and 0.94 (95% confidence intervals low bounds (95% CI(low)) were 0.66 and 0.82), respectively. To study the reliability of beta-sitosterol and campesterol measurement over time, seven premenopausal women were recruited. Over a 6-month period, each woman provided a fasting blood sample once a month at the same time of day, and the same numerical day of the luteal phase of her menstrual cycle (between the 20th and 24th day of her menstrual cycle). All plasma samples from the same individual were processed together at the same time by the same technician at the end of the 6-month period. The overtime ICCs of plasma beta-sitosterol and campesterol were 0.91 (95% CI(low) 0.49) and 0.58 (95% CI(low) 0.31), respectively. The high reproducibility and good overtime reliability of plasma beta-sitosterol and campesterol measurements indicate that they may be suitable for potential clinical and population-based studies on cancer prevention. PMID- 11432712 TI - Screening with faecal occult blood test (FOBT) for colorectal cancer: assessment of two methods that attempt to improve compliance. AB - Screening with the faecal occult blood test (FOBT) has been shown in randomized control trials to be effective in reducing mortality from colorectal cancer. Compliance to this test recommendation, however, by the general population is usually low. To evaluate different methods of increasing compliance with FOBT, using mailed test kits or order cards, with or without information leaflets, subjects were randomly assigned to receive a test kit or a kit request card. An information leaflet was included in half of the mailings. All participants were contacted for interview. Compliance was evaluated through the central computer system of the study's FOBT laboratory. Self-initiated compliance with FOBT in the year preceding the study was 0.6% of the study participants. The overall compliance rate with the programme invitation was 17.9%, with a somewhat higher, though non-significant response to the mailed kit (19.9%) over the kit request card (15.9%). Women complied with the test significantly more than men, older participants more than younger. Compliance to FOBT is low among the Israeli population aged 50-74 who receive a formal invitation to carry out this screening. Mailing a kit request card within the framework of a screening programme can achieve a substantial increase (to 17.9%) in the level of compliance for the relatively low cost of postage. More effort is needed to study additional means of convincing the non-responders to take part in this potentially life saving activity. PMID- 11432713 TI - An intervention trial to inhibit the progression of precancerous gastric lesions: compliance, serum micronutrients and S-allyl cysteine levels, and toxicity. AB - Gastric cancer is the second most frequent cause of death from cancer in the world and the leading cause of death from cancer in China. In September 1995, we launched a randomized multi-intervention trial to inhibit the progression of precancerous gastric lesions in Linqu County, Shandong Province, an area of China with one of the world's highest rates of gastric cancer. Treatment compliance was measured by pill counts and quarterly serum concentrations of vitamin C, vitamin E and S-allyl cysteine. In 1999, toxicity information was collected from each trial participant to evaluate treatment-related side-effects during the trial. Compliance rates were 93% and 92.9% for 39 months of treatment with the vitamins/mineral and garlic preparation, respectively. The means for serum concentrations of vitamins C and E were 7.2 microg/ml and 1695 microg/dl among subjects in the active treatment groups compared with 3.1 microg/ml and 752 microg/dl among subjects in the placebo treatment group, respectively. No significant differences in side-effects were observed between the placebo treatment group and the vitamins/mineral and garlic preparation treatment groups during the 39-month trial period. PMID- 11432714 TI - Determination of N-nitrosodimethylamine in artificial gastric juice by gas chromatography-mass spectrometry and by gas chromatography-thermal energy analysis. AB - The thermal energy analyser (TEA) is considered to be the gold standard for the determination of nitrosamines. However, since many laboratories cannot justify the use of such a very specific detection system, alternative detection methods are useful. While standard gas chromatography (GC) detectors lack the selectivity of the TEA detector, mass spectrometry (MS) seems to be the method of choice to combine GC separation with mass selective detection. Moreover, the detection limits of the GC-MS assay in general use are about 4 times lower than those of the GC-TEA assay. A comparison of GC-MS and GC-TEA data on N-nitrosodimethylamine determinations showed a strong correlation between the two assays (R2 = 0.86), demonstrating the exchangeability of these methods. PMID- 11432715 TI - Interval cancers in a French breast cancer-screening programme (Somme Department). AB - The objective of this study is to analyse the detection rates and tumour diameter of interval cancers in the breast cancer mass-screening programme of Somme Department (France), launched in 1990. Interval cancers are defined as breast cancers diagnosed within 36 months after a negative screening assessment, for women attending the programme between December 1990 to December 1993. Age adjusted incidence rates were 0.51 per 1000 woman-years of follow-up in the 3 year interval after initial and subsequent screens. Diagnosis is made at early stage (sizes < or = 10 mm) in 20% of interval cancers. This stage is higher than that in screened women (9% of in situ cases and 35% of very small tumours). Interval cancer rates are low during the first year (0.18 per 1000 woman-years of follow-up) but higher in the second and third years. PMID- 11432716 TI - Testosterone and oestradiol in relation to tobacco smoking, body mass index, energy consumption and nutrient intake among adult men. AB - There is substantial evidence linking steroid hormones and diet to cancer aetiology. The evidence on lifestyle determinants of steroid hormones, however, is limited. We have conducted a study to identify dietary and other lifestyle predictors of testosterone and oestradiol among adult men. Subjects were 112 healthy Greek men, recruited as controls in a case-control study on the aetiology of liver cancer. Demographic data and detailed histories of smoking habits and alcohol consumption were recorded. Diet was assessed through an interviewer administered validated food-frequency questionnaire. Serologic measurements of oestradiol, testosterone and sex hormone binding globulin were also conducted. We developed linear regression models to evaluate the associations of smoking and dietary factors with serum testosterone and oestradiol. The results indicate that, among men, both testosterone and oestradiol serum levels decline with age, whereas body mass index may be inversely related with testosterone and positively with oestradiol. The evidence concerning alcohol in relation to these hormones is inconclusive. Emerging evidence concerning smoking suggests positive associations with both hormones in the blood. The principal nutritional findings are a positive association of carbohydrate intake with testosterone levels and a set of inverse associations of vitamins with oestradiol. PMID- 11432717 TI - Breast cancer incidence in ex-smokers in relation to body mass index, weight gain and blood lipid levels. AB - According to several studies breast cancer is more common among former smokers. This study explores whether this association has any relationship with anthropometric measurements or blood lipid levels. The 2082 ex-smokers (mean age 49.9 years) in the Malmo Preventive Cohort were followed for an average of 13.3 years using official cancer registries. This yielded 93 incident breast cancer cases. Oestrogen receptor (ER) status was assessed by an immunological method. Incidence of breast cancer covaried with height, body mass index, weight gain and cholesterol levels. None of these associations reached statistical significance. Incidence of breast cancer increased over quartiles of serum triglycerides, Ptrend: 0.02, relative risk (RR) for triglycerides as a continuous variable: 1.46 (1.21-1.77). Nineteen tumours were ER negative; this subgroup was similarly related to high triglycerides, 1.76 (1.40-2.21). All results were similar when BMI and cholesterol levels were entered into the model. It is concluded that breast cancer incidence covaries with triglyceride levels in ex-smokers. PMID- 11432718 TI - Readiness to change sun-protective behaviour. AB - The incidence of malignant melanoma and non-melanoma skin cancers has increased rapidly in Sweden during the last 20 years. The best-known way to revert this trend is primary prevention. Matching health messages to readiness to change in the population may enhance the effect of community-based prevention. The aims of this study were to investigate readiness to change sun-protective behaviour in two groups (visitors to mobile screening units and beach-goers) and to test a single-item algorithm in assessing the stage of change in sun-protective behaviour. Seven hundred and forty-two visitors to the mobile screening units and 202 individuals on nearby beaches answered a short questionnaire. The assessment of readiness to change was based on stages of change in sun-protective behaviour modified from the Transtheoretical Model of Behaviour Change. As expected, the visitors to the screening units were more often in action/maintenance stages than the beach group for most sun-protective behaviours. In conclusion, the single item algorithm method appears to be sensitive to assess readiness to change sun protective behaviour, based on the Transtheoretical Model of Behaviour Change. This method can be incorporated into population surveys and may aid in developing successful skin cancer prevention programmes. PMID- 11432719 TI - Coke is not it. PMID- 11432720 TI - Aetiology of sex differences in colorectal cancer. PMID- 11432721 TI - Cascades of transcriptional induction during human lymphocyte activation. AB - Lymphocyte activation is known to be associated with the induction of genes implicated in cytokine signaling and cellular proliferation. High-density microarrays offer the means to monitor global cellular expression profiles, temporal relationships between classes of transcripts, and alterations associated with human disease or immunosuppression. We sought to determine whether microarray analysis would accurately reflect the normal pattern of gene expression following human T cell activation, and whether the complex expression patterns identified could be analyzed to produce a functional profile of lymphocyte activation. We examined a time course of sequential expression profiles for 6,800 cellular transcripts in human lymphocytes activated with concanavalin A. Expression patterns were grouped using clustering analysis and validated using Northern blotting. Genes known to be induced following T cell activation were accurately identified, and the qualitative patterns of gene expression were well correlated between Northern and microarray analyses. Quantitative differences in gene expression levels were less well correlated between these two techniques. Expression profile analysis revealed the sequential induction of groups of functionally similar genes, whose temporal coregulation underscores known cellular events during T cell activation. This functional "fingerprint" of lymphocyte activation may prove useful for comparisons of lymphocyte responses under experimental conditions and in disease states. PMID- 11432722 TI - Induction of basic helix-loop-helix protein DEC1 (BHLHB2)/Stra13/Sharp2 in response to the cyclic adenosine monophosphate pathway. AB - DEC1 (BHLHB2)/Stra13/Sharp2, a basic helix-loop-helix (bHLH) transcription factor has been suggested to be involved in the control of proliferation and/or differentiation of several cells including nerve cells, fibroblasts and chondrocytes. In the present study, we examined the effect of parathyroid hormone (PTH), dibutyryl cAMP (Bt2cAMP) and forskolin on the expression of DEC1 in various cells. In rabbit chondrocyte cultures, PTH or Bt2cAMP increased the DEC1 mRNA level within 1 h. Thereafter, the DEC1 mRNA level rapidly decreased to the basal level at 3 h, and increased at 6-24 h. In cultures of a mouse embryo prechondrogenic cell line ATDC5, PTH or forskolin, an activator of adenylate cyclase, also increased the DEC1 mRNA level within 1 h. Furthermore, in all evaluated cell lines of human fibroblasts, canine epithelial cells, human carcinoma, human glioblastoma and human melanoma, Bt2cAMP increased the DEC1 mRNA level within 1-3 h. Studies with actinomycin D and cycloheximide indicated that the enhancement of DEC1 mRNA by cAMP was not due to mRNA stabilization and did not require new protein synthesis. These findings suggest that DEC1 is a novel direct target for cAMP in wide types of cells, and that the bHLH protein is involved in the control of gene expression in cAMP-activated cells. PMID- 11432723 TI - A combination of MIP-3alpha and TGF-beta1 is required for the attraction of human Langerhans precursor cells through a dermal-epidermal barrier. AB - Signals regulating the traffic of Langerhans cell precursors from blood to the epidermis are not yet fully understood. The observations that TGF-beta1 is of unique importance in Langerhans cells (LC) ontogeny and that macrophage inflammatory protein-3alpha (MIP-3alpha) is able to attract LC within the epidermis, prompted us to study the effect of MIP-3alpha and TGF-beta1 on the migration of LC precursors. The migratory capacity of immature dendritic cells (DC) was assessed using a reconstituted basement membrane assay (Matrigel), mimicking the prerequisite passage through the dermal-epidermal basement membrane on the way into the epidermis. DC differentiated from cord blood CD34 cells in the presence of GM-CSF plus TNF-alpha were subjected to migration using modified Boyden chambers. Day-6 DC progenitors migrated in a dose-dependent fashion in response to MIP-3alpha, and CD1alpha+ LC precursors responded preferentially to the chemokine. Immature DC did not respond strongly to TGF-beta1 alone in migration assays, but up to 68% of the cells migrated in response to MIP-3alpha plus TGF-beta1. Among them, at least 50% expressed CD1a and E-cadherin and can be considered LC precursors. The allostimulatory function of these cells was significantly more potent than that which migrated in response to MIP-3alpha alone. Our results show that a significant proportion of immature DC is able to migrate through a dermal-epidermal basement membrane equivalent. In the presence of TGF-beta1, the DC which respond to MIP-3alpha have the phenotype and the functional capacity of epidermal LC. Our findings underline the role of MIP 3alpha and TGF-beta1 in attraction and localization of immature LC within the epidermis under normal conditions. PMID- 11432724 TI - Intracellular location and nuclear targeting of the Spi-1, Spi-2 and Spi-3 gene derived serine protease inhibitors in non-secretory cells. AB - Proteases and their inhibitors are indispensable for the regulated activation and/or degradation of structural and functional proteins involved in basic cellular processes, e.g. in cell cycle control, cell growth, differentiation and apoptosis. In this context the serine protease inhibitors derived from the murine Spi-1, Spi-2 and Spi-3 genes, and their human homologs, deserve reconsideration. Microsequencing data indicate that a fraction of the three serpins has the capability to constitute a well characterized proteinase K, high salt and SDS stable complex which coisolates with DNA under salting out conditions from various cell and tissue types. This tight association with DNA isolated under conditions designed to deproteinize DNA efficiently points to an in situ preformed chromatin complex. Accordingly, in addition to their well known functions as 'serum protease inhibitors' the Spi-1 and Spi-2 gene-derived proteins appear to have intracellular functions as well. The involvement of the three serpins in chromatin complexes requires their nuclear translocation. Application of (enhanced) green fluorescent protein technology and optical section microscopy reveals that truncation of the N-terminal signal sequences of the Spi-1 and Spi-2 gene-encoded proteins is a prerequisite for their nuclear translocation while non-truncated fusion proteins are enriched at the nuclear indentation which is the site of the Golgi apparatus and the centrosome. The identification of new species of intracellular serpins is of potential interest with respect to accumulating evidence for serine protease inhibitor-dependent inhibition or prevention of apoptosis. PMID- 11432725 TI - Functional analysis of the sheep Wilson disease protein (sATP7B) in CHO cells. AB - In this study we investigated the function of the sheep orthologue of ATP7B (sATP7B), the protein affected in the human copper toxicosis disorder Wilson disease. Two forms of sATP7B are found in the sheep, a 'normal' form and one with an alternate N terminus, both of which were expressed in CHO-K1 cells. Cells expressing either form of sATP7B were more resistant to copper than the parental CHO-K1 cells. Subcellular localisation studies showed that both forms of sATP7B were similarly located in the trans-Golgi network (TGN). When the extracellular copper concentration was increased, each form of sATP7B redistributed to a punctate, vesicular compartment that extended throughout the cytoplasm. Both forms of sATP7B recycled to the perinuclear location within one hour when the cells were subsequently incubated in basal medium. After treatment of cells with bafilomycin A1 sATP7B accumulated in cytoplasmic vesicles, implying that ATP7B continuously recycles via the endocytic pathway. These results suggest that both forms of sATP7B are functional copper-transport proteins and that the intracellular location and trafficking of the sheep protein within the cell also appears normal. PMID- 11432726 TI - Co-localization of vesicles and P/Q Ca2+-channels explains the preferential distribution of exocytotic active zones in neurites emitted by bovine chromaffin cells. AB - We have taken advantage of the differences between the preferential localization of secretion in the terminals of neurite-emitting bovine chromaffin cells in contrast with the random distribution secretion in spherical cells to study the possible molecular factors determining such localization by using immunofluorescence and confocal microscopy techniques. By analyzing the distribution of dopamine beta-hydroxylase present in the membrane of chromaffin granules, we found that vesicles migrate and accumulate in dense packages in the terminals of neurite processes. Neither members of the fusion core complex such as SNAP-25, nor nicotinic receptors are preferentially located in the terminals as would be expected from elements defining sites of release, thereby suggesting the presence of additional factors. Interestingly, we observed a preferential distribution of the P/Q subtype of Ca2+ channels in these neurite terminals and co-localization with vesicles present in these structures, in sharp contrast with the overall distribution of the L subtype channels. Using the same immunofluorescence techniques we were unable to detect N-type calcium channels. In addition, omega-agatoxin IVA was able to block 70% of the exocytotic release occurring into the neurites, whereas L-type blockers had a weak effect. Taken together our results strongly indicate that the co-localization of vesicles and clusters of P/Q Ca2+ channels may explain the precise localization of exocytotic sites in the terminals of neurite-emitting chromaffin cells, whereas the distribution of secretory sites in round cells may arise from the random presence of these factors as indicated by their partial co-localization. PMID- 11432727 TI - Serum withdrawal induces a redistribution of intracellular gelsolin towards F actin in NIH 3T3 fibroblasts preceding apoptotic cell death. AB - The intracellular distribution of gelsolin in NIH 3T3 cells was examined by immunostaining using affinity-purified polyclonal gelsolin antibodies before and after induction of apoptosis by serum withdrawal. Serum deprivation induced detachment of an increasing number of NIH 3T3 cells, but also apoptosis in attached cells as verified morphologically by chromatin condensation, nuclear fragmentation and labelling of their periphery by FITC-annexin V. Ongoing apoptosis was also demonstrated by activation of caspase-3 activity and chromatin cleavage into high-molecular-mass fragments, although no internucleosomal chromatin degradation (DNA-ladder formation) was detected. When cells were maintained in the presence of 10% foetal calf serum, gelsolin immunoreactivity was evenly distributed in the cytoplasm. No obvious co-localisation of gelsolin and the actin-containing stress fibres was detected under these conditions. At day one after serum withdrawal, a redistribution of gelsolin to actin filaments was detected within a few attached cells by double fluorescence staining. The number of cells exhibiting this redistribution increased at days two to four. In addition, the stress fibres increased in thickness and their length was continuously reduced. At day four, many cells contained shortened stress fibres, which had lost their longitudinal orientation. Additionally, the cytoplasm of a number of attached cells was highly condensed around their nuclei and a homogenous distribution of both gelsolin and actin was detected in the remaining cytoplasmic rim. Up to day two, these effects were reversible after re-addition of serum to attached cells. A similar redistribution of gelsolin immunore activity was observed after induction of apoptosis by cycloheximide, but not after initiation of necrosis by hydrogen peroxide. In NIH 3T3 cells no alteration in the expression of gelsolin at the level of protein (Western blot) or specific mRNA (Northern blot) was observed after serum withdrawal. Using Western blotting, no proteolysis of gelsolin was detected up to day 4, although caspase-3 activity was found to have increased fivefold after serum withdrawal. These results suggested that in these cells F-actin severing might occur in the absence or advance of gelsolin cleavage by caspases. Intact gelsolin on its own may be sufficient for the dissolution of the microfilaments, since micro-injection of gelsolin into primary bovine lens cells led to a transient disappearance of the stress fibres and to a reduction of their attachment area to the substratum. In NIH 3T3 cells similar effects of micro-injected gelsolin were only observed at day one after serum withdrawal. PMID- 11432728 TI - The drug/metabolite transporter superfamily. AB - Previous work defined several families of secondary active transporters, including the prokaryotic small multidrug resistance (SMR) and rhamnose transporter (RhaT) families as well as the eukaryotic organellar triose phosphate transporter (TPT) and nucleotide-sugar transporter (NST) families. We show that these families as well as several other previously unrecognized families of established or putative secondary active transporters comprise a large ubiquitous superfamily found in bacteria, archaea and eukaryotes. We have designated it the drug/metabolite transporter (DMT) superfamily (transporter classification number 2.A.7) and have shown that it consists of 14 phylogenetic families, five of which include no functionally well-characterized members. The largest family in the DMT superfamily, the drug/metabolite exporter (DME) family, consists of over 100 sequenced members, several of which have been implicated in metabolite export. Each DMT family consists of proteins with a distinctive topology: four, five, nine or 10 putative transmembrane alpha helical spanners (TMSs) per polypeptide chain. The five TMS proteins include an N-terminal TMS lacking the four TMS proteins. The full-length proteins of 10 putative TMSs apparently arose by intragenic duplication of an element encoding a primordial five-TMS polypeptide. Sequenced members of the 14 families are tabulated and phylogenetic trees for all the families are presented. Sequence and topological analyses allow structural and functional predictions. PMID- 11432729 TI - Caenorhabditis elegans expresses a functional phytochelatin synthase. AB - The formation of phytochelatins, small metal-binding glutathione-derived peptides, is one of the well-studied responses of plants to toxic metal exposure. Phytochelatins have also been detected in some fungi and some marine diatoms. Genes encoding phytochelatin synthases (PCS) have recently been cloned from Arabidopsis, wheat and Schizosaccharomyces pombe. Surprisingly, database searches revealed the presence of a homologous gene in the Caenorhabditis elegans genome, DDBJ/EMBL/GenBank accession no. 266513. Here we show that C. elegans indeed expresses a gene coding for a functional phytochelatin synthase. CePCS complements the Cd2+ sensitivity of a Schizosaccharomyces pombe PCS knock-out strain and confers phytochelatin synthase activity to these cells. Thus, phytochelatins may play a role for metal homeostasis also in certain animals. PMID- 11432730 TI - Monitoring of secondary and tertiary structure changes in the gastric H+/K+ ATPase by infrared spectroscopy. AB - Conformational changes occurring in the catalytic cycle of the H+/K+-ATPase were monitored by Fourier transform infrared spectroscopy (FTIR). Caged compounds were used to release ATP, in the presence of Ca2+, to induce the transition between the E1 and E1-P conformation of the H+/K+-ATPase. In addition to bands associated with the photolysis of the caged compounds, some peaks of the difference infrared spectra were associated with changes in secondary structure and modifications of the ionization in the side chains of amino-acid residues (Glu or Asp). These changes were specific to the reaction between the ligand and the enzyme. We estimated that 39 amino acids changed their secondary structure during the reaction and four amino-acid residues were deprotonated. Similar spectral changes appeared when ADP was released from its precursor. The release of Pi from the same caged molecule did not induce similar changes. Changes in tertiary structure occurring during the binding of adenosine and phosphorylation of the enzyme were demonstrated by recording hydrogen/deuterium exchange kinetics by attenuated total reflectance FTIR spectroscopy (ATR-FTIR). At least 129 amide protons were involved in a tertiary structure change induced by ATP. This suggested that secondary structure change transduced a much larger tertiary structure modification. PMID- 11432731 TI - S1 proteins C2 and D2 are novel hnRNPs similar to the transcriptional repressor, CArG box motif-binding factor A. AB - S1 proteins A-D are liberated from thoroughly washed nuclei by mild digestion with DNase I or RNase A, and extracted selectively at pH 4.9 from the reaction supernatants. Here, we characterized the S1 proteins, focusing on protein D2, the most abundant S1 protein in the rat liver, and on protein C2 as well. Using a specific antibody, McAb 351, they were shown to occur in the extranucleolar nucleoplasm, and to be extracted partly in the nuclear soluble fraction. We demonstrate that the S1 proteins in this fraction exist constituting heterogeneous nuclear ribonucleoproteins (hnRNPs), through direct binding to hnRNAs, as revealed by centrifugation on density gradients, immunoprecipitation, and UV cross-linking. In hnRNPs, protein D2 occurred at nuclease-hypersensitive sites and C2 in the structures that gave rise to 40 S RNP particles. By microsequencing, protein D2 was identified with a known protein, CArG box motif binding factor A (CBF-A), which has been characterized as a transcriptional repressor, and C2 as its isoform protein. In fact, CBF-A expressed from its cDNA was indistinguishable from protein D2 in molecular size and immunoreactivity to McAb 351. Thus, the present results demonstrate that S1 proteins C2 and D2 are novel hnRNP proteins, and suggest that the proteins C2 and D2 act in both transcriptional and post-transcriptional processes in gene expression. PMID- 11432732 TI - Photoactivatable alpha-conotoxins reveal contacts with all subunits as well as antagonist-induced rearrangements in the Torpedo californica acetylcholine receptor. AB - Azidobenzoyl (AzBz) and benzoylbenzoyl (BzBz) derivatives of alpha-conotoxin MI and L-benzoylphenylalanine (Bpa) analogs of alpha-conotoxin GI were synthesized. All these compounds, similarly to native alpha-conotoxins, completely displaced the radioiodinated MI or GI from the membrane-bound nicotinic acetylcholine receptor (AChR) of Torpedo californica. However, the GI(Bpa11) analog was considerably less potent than GI in competing with radioiodinated alpha bungarotoxin (alphaBgt). Irradiation of iodinated AzBz derivatives bound to AChR resulted in labeling of all AChR subunits. The BzBz and Bpa derivatives gave lower levels of specific cross-linking but considerable labeling at additional sites that was enhanced, rather than suppressed, by an excess of native alpha conotoxins or alphaBgt. Both equilibrium binding of benzophenone-derivatized alpha-conotoxins and their cross-linking could be totally abolished by physostigmine. The results obtained demonstrate that (a) specific binding sites for alpha-conotoxins and alphaBgt are overlapping but not identical, (b) each of the AChR subunits can be labeled with photoactivatable alpha-conotoxins and (c) enhancement of benzophenone-derivatized alpha-conotoxins cross-linking at additional (physostigmine-related) sites by alphaBgt or GI indicates that these antagonists induce structural alterations in the AChR outside their binding sites. PMID- 11432733 TI - Mechanism of activation of the double-stranded-RNA-dependent protein kinase, PKR: role of dimerization and cellular localization in the stimulation of PKR phosphorylation of eukaryotic initiation factor-2 (eIF2). AB - An important defense against viral infection involves inhibition of translation by PKR phosphorylation of the alpha subunit of eIF2. Binding of viral dsRNAs to two dsRNA-binding domains (dsRBDs) in PKR leads to relief of an inhibitory region and activation of eIF2 kinase activity. Interestingly, while deletion of the regulatory region of PKR significantly induces activity in vitro, the truncated kinase does not inhibit translation in vivo, suggesting that these sequences carry out additional functions required for PKR control. To delineate these functions and determine the order of events leading to activation of PKR, we fused truncated PKR to domains of known function and assayed the chimeras for in vivo activity. We found that fusion of a heterologous dimerization domain with the PKR catalytic domain enhanced autophosphorylation and eIF2 kinase function in vivo. The dsRBDs also mediate ribosome association and we proposed that such targeting increases the localized concentration of PKR, enhancing interaction between PKR molecules. We addressed this premise by linking the truncated PKR to RAS sequences mediating farnesylation and membrane localization and found that the fusion protein was functional in vivo. These results indicate that cellular localization along with oligomerization enhances interaction between PKR molecules. Alanine substitution for the phosphorylation site, threonine 446, impeded in vivo and in vitro activity of the PKR fusion proteins, while aspartate or glutamate substitutions partially restored the function of the truncated kinase. These results indicate that both dimerization and cellular localization play a role in transient protein-protein interactions and that trans autophosphorylation is the final step in the mechanism of activation of PKR. PMID- 11432734 TI - Crystal structure of Fab198, an efficient protector of the acetylcholine receptor against myasthenogenic antibodies. AB - The crystal structure of the Fab fragment of the rat monoclonal antibody 198, with protective activity for the main immunogenic region of the human muscle acetylcholine receptor against the destructive action of myasthenic antibodies, has been determined and refined to 2.8 A resolution by X-ray crystallographic methods. The mouse anti-lysozyme Fab D1.3 was used as a search model in molecular replacement with the AMORE software. The complementarity determining regions (CDR)-L2, CDR-H1 and CDR-H2 belong to canonical groups. Loops CDR-L3, CDR-H2 and CDR-H3, which seem to make a major contribution to binding, were analyzed and residues of potential importance for antigen-binding are examined. The antigen binding site was found to be a long crescent-shaped crevice. The structure should serve as a model in the rational design of very high affinity humanized mutants of Fab198, appropriate for therapeutic approaches in the model autoimmune disease myasthenia gravis. PMID- 11432735 TI - The physico-chemical characteristics of the phosphocholine-containing glycoglycerolipid MfGL-II govern the permeability properties of Mycoplasma fermentans. AB - Mycoplasma fermentans seems to be involved in several pathogenic conditions in humans, and is among other things capable of fusing with T-cells and lymphocytes. The choline-containing phosphoglycolipid 6'-O-(3"-phosphocholine-2"-amino-1" phospho-1",3"-propanediol)-alpha-D-glucopyranosyl-(1'-->3)-1,2-diacylglycerol (MfGL-II) in the membrane of M. fermentans has been suggested to enhance the fusion process, and the characteristics of MfGL-II were therefore investigated. When a cell culture ages the fraction of MfGL-II increases, and the fraction of the other major membrane lipid, phosphatidylglycerol (PtdGro), decreases concomitantly. Swelling experiments showed that the permeability and osmotic fragility are markedly reduced in aged cells. MfGL-II is selectively released into the surrounding medium when aged M. fermentans cells are incubated in buffer containing EDTA. The physico-chemical properties of MfGL-II were studied by NMR spectroscopy and differential scanning calorimetry, and they can explain the biochemical results. The temperature for the transition between gel and lamellar liquid crystalline (Lalpha) phases is 35-45 degrees C higher for MfGL-II than for PtdGro, which most probably gives rise to the reduced permeability in aged cells. At high water contents MfGL-II forms an Lalpha phase and isotropic aggregates which were interpreted to be vesicles with a radius of approximately 450 A. It is proposed that MfGL-II forms vesicles in the surrounding medium when it is released from the cell membrane. Neither EDTA nor Ca2+ ions have a significant influence on the aggregate structures formed by MfGL-II. Our results indicate that MfGL-II has no fusogenic properties. It is more probable that a recently identified lysolipid in the M. fermentans membrane acts as a fusogen. PMID- 11432736 TI - Expression and functional analysis of a gene cluster involved in the synthesis of decaprenoxanthin reveals the mechanisms for C50 carotenoid formation. AB - Corynebacterium glutamicum accumulates the C50 carotenoid decaprenoxanthin. Rescued DNA from transposon color mutants of this Gram-positive bacterium was used to clone the carotenoid biosynthetic gene cluster. By sequence comparison and functional complementation, the genes involved in the synthesis of carotenoids with 50 carbon atoms were identified. The genes crtE, encoding a geranylgeranyl pyrophosphate synthase, crtB, encoding a phytoene synthase, and crtI, encoding a phytoene desaturase, are responsible for the formation of lycopene. The products of three novel genes, crtYe and crtYf, with sequence similarities to heterodimeric lycopene cyclase crtYc and crtYd, together with crtEb which exhibits a prenyl transferase motif, were involved in the conversion of C40 acyclic lycopene to cyclic C50 carotenoids. Using functional complementation in Escherichia coli, it could be shown that the elongation of lycopene to the acyclic C50 carotenoid flavuxanthin by the addition of C5 isoprenoid units at positions C-2 and C-2' is catalyzed by the crtEb gene product. Subsequently, the gene products of crtYe and crtYf in a concerted action convert the acyclic flavuxanthin into the cyclic C50 carotene, decaprenoxanthin, forming two epsilon-ionone groups. The mechanisms, involving two individual steps for the formation of cyclic C50 carotenoids from lycopene, are proposed on the basis of these results. PMID- 11432737 TI - Properties of the C-terminal domain of 4.1 proteins. AB - At the C-terminus of all known 4.1 proteins is a sequence domain unique to these proteins, known as the C-terminal domain (CTD). Mammalian CTDs are associated with a growing number of protein-protein interactions, although such activities have yet to be associated with invertebrate CTDs. Mammalian CTDs are generally defined by sequence alignment as encoded by exons 18-21. Comparison of known vertebrate 4.1 proteins with invertebrate (Caenorhabditis elegans and Drosophila melanogaster) 4.1 proteins indicates that mammalian 4.1 exon 19 represents a vertebrate adaptation that extends the sequence of the CTD with a Ser/Thr-rich sequence. The CTD was first described as a 22/24-kDa domain by chymotryptic digestion of erythrocyte 4.1 (4.1R) [Leto, T.L. & Marchesi, V.T. (1984) J. Biol. Chem. 259, 4603-4608]. Here we show that in 4.1R the 22/24-kDa fragment is not stable but rapidly processed to a 15-kDa fragment by chymotrypsin. The 15-kDa fragment is extremely stable, being resistant to overnight digestion in chymotrypsin on ice. Analysis of this fragment indicates that it is derived from residues 709-858 (SwissProt accession no. P48193), and represents the CTD of 4.1R. The fragment behaves as a globular monomer in solution. Secondary-structure predictions indicate that this domain is composed of five or six beta strands with an alpha helix before the most C-terminal of these. Together these data indicate that the CTD probably represents an independent folding structure which has gained function since the divergence of vertebrates from invertebrates. PMID- 11432738 TI - Cloning and sequencing of a molluscan endo-beta-1,4-glucanase gene from the blue mussel, Mytilus edulis. AB - Using polymerase chain reaction, cloning and sequencing techniques, a complementary DNA encoding a low molecular mass cellulase (endo-1,4-beta-D glucanase, EC 3.2.1.4) has been identified in the digestive gland of the marine mussel, Mytilus edulis. It contains a 5' untranslated region, a 633-nucleotide ORF encoding a 211 amino-acid protein, including a 17 amino-acid signal peptide and a complete 3' untranslated region. At the C-terminal end of the purified mature protein, a 13 amino-acid peptide is lacking in comparison to the protein sequence deduced from the ORF. This peptide is probably removed as a consequence of post-translational amidation of the C-terminal glutamine. The endoglucanase genes have been isolated and sequenced from both Swedish and French mussels. The coding parts of these two sequences are identical. Both genes contain two introns, the positions of which are conserved. However the length of the introns are different due to base substitutions, insertions or deletions showing the existence of interspecies length polymorphism. The percentage of similarity for the introns of the two gene sequences is 96.9%. This is the first time a molluscan cellulase is characterized at DNA level. Amino acid sequence-based classification has revealed that the enzyme belongs to the glycosyl hydrolase family 45 [B. Henrissat (Centre de Recherches sur les Macromolecules Vegetales, CNRS, Joseph Fourier Universite, Grenoble, France), personal communication]. There is no cellulose binding domain associated with the sequence. PMID- 11432739 TI - Modulation of the lipid binding properties of the N-terminal domain of human apolipoprotein E3. AB - Apolipoprotein E (apoE) plays a critical role in plasma lipid homeostasis through its function as a ligand for the low-density lipoprotein (LDL) receptor family. Receptor recognition is mediated by residues 130-150 in the independently folded, 22-kDa N-terminal (NT) domain. This elongated globular four-helix bundle undergoes a conformational change upon interaction with an appropriate lipid surface. Unlike other apolipoproteins, apoE3 NT failed to fully protect human LDL from aggregation induced by treatment with phospholipase C. Likewise, in dimyristoylglycerophosphocholine (Myr2Gro-PCho) vesicle transformation assays, 100 microg apoE3 NT induced only 15% reduction in vesicle (250 microg) light scattering intensity after 30 min. ApoE3 NT interaction with modified lipoprotein particles or Myr2Gro-PCho vesicles was concentration-dependent whereas the vesicle transformation reaction was unaffected by buffer ionic strength. In studies with the anionic phospholipid dimyristoylglycerophosphoglycerol, apoE3 NT mediated vesicle transformation rates were enhanced > 10-fold compared with Myr2Gro-PCho and activity decreased with increasing buffer ionic strength. Solution pH had a dramatic effect on the kinetics of apoE3 NT-mediated Myr2Gro PCho vesicle transformation with increased rates observed as a function of decreasing pH. Fluorescence studies with a single tryptophan containing apoE3 NT mutant (L155W) revealed increased solvent exposure of the protein interior at pH values below 4.0. Similarly, fluorescent dye binding experiments with 8-anilino-1 naphthalene sulfonate revealed increased exposure of apoE3 NT hydrophobic interior as a function of decreasing pH. These studies indicate that apoE3 NT lipid binding activity is modulated by lipid surface properties and protein tertiary structure. PMID- 11432740 TI - SP3 and AP-1 mediate transcriptional activation of the lamin A proximal promoter. AB - Lamin A is a major component of the nuclear lamina that is expressed in various types of differentiated cells. We have analysed previously the putative promoter sequences of the gene and shown that the rat lamin A proximal promoter contains two essential motifs, a GC box that can bind to Sp1 and Sp3, and an AP-1 motif that can bind to c-Jun and c-Fos. In this study we have investigated the role of Sp1 and Sp3 in transactivation of the promoter. Functional analysis of the promoter in Drosophila SL2 cells has demonstrated that it is inactive in the absence of Sp proteins. Activation by expression of Sp3 is more pronounced than that by Sp1 although both proteins can bind to the GC box in vitro; activation clearly depends on an intact GC box as deduced from mutant analysis. Promoter activity in SL2 cells also requires an intact AP-1 motif, which can bind to endogenous Drosophila Jun and Fos proteins. Furthermore, overexpression of c-Jun and c-Fos results in fourfold activation of the promoter in PCC-4 embryonal carcinoma cells. Our demonstration that activation of the lamin A proximal promoter is mediated by Sp3 and AP-1 transcription factors affords a basis for further studies on the regulation of this important gene during development and disease. PMID- 11432741 TI - Overproduction of a functional A1 ATPase from the archaeon Methanosarcina mazei Go1 in Escherichia coli. AB - Single subunits of the A1 ATPase from the archaeon Methanosarcina mazei Go1 were produced in E. coli as MalE fusions and purified, and polyclonal antibodies were raised against the fusion proteins. A DNA fragment containing the genes ahaE, ahaC, ahaF, ahaA, ahaB, ahaD, and ahaG, encoding the hydrophilic A1 domain and part of the stalk of the A1AO ATPase of M. mazei Go1, was constructed, cloned into an expression vector and transformed into different strains of Escherichia coli. In any case, a functional, ATP-hydrolysing A1 ATPase was produced. Western blots demonstrated the production of subunits A, B, C, and F in E. coli, and minicell analyses suggested that subunits D, E, and G were produced as well. This is the first demonstration of a heterologous production of a functional ATPase from an archaeon. The A1 ATPase was sensitive to freezing but lost only about 50% of its activity within 18 days on ice. Inhibitor studies revealed that the heterologously produced A1 ATPase is insensitive to azide, dicyclohexylcarbodiimide and bafilomycin A1, but sensitive to diethylstilbestrol and its analogues dienestrol and hexestrol. The expression system described here will open new avenues towards the functional and structural analyses of this unique class of enzymes. PMID- 11432742 TI - Analysis of Usp DNA binding domain targeting reveals critical determinants of the ecdysone receptor complex interaction with the response element. AB - The steroid hormone, 20-hydroxyecdysone (20E), directs Drosophila metamorphosis via a heterodimeric receptor formed by two members of the nuclear hormone receptors superfamily, the product of the EcR (EcR) and of the ultraspiracle (Usp) genes. Our previous study [Niedziela-Majka, A., Kochman, M., Ozyhar, A. (2000) Eur. J. Biochem. 267, 507-519] on EcR and Usp DNA-binding domains (EcRDBD and UspDBD, respectively) suggested that UspDBD may act as a specific anchor that preferentially binds the 5' half-site of the pseudo-palindromic response element from the hsp27 gene promoter and thus locates the heterocomplex in the defined orientation. Here, we analyzed in detail the determinants of the UspDBD interaction with the hsp27 element. The roles of individual amino acids in the putative DNA recognition alpha helix and the roles of the base pairs of the UspDBD target sequence have been probed by site-directed mutagenesis. The results show how the hsp27 element specifies UspDBD binding and thus the polar assembly of the UspDBD/EcRDBD heterocomplex. It is suggested how possible nucleotide deviations within the 5' half-site of the element may be used for the fine-tuning of the 20E-response element specificity and consequently the physiological response. PMID- 11432743 TI - Novel polyketides synthesized with a higher plant stilbene synthase. AB - The physiological function of the stilbene synthase (STS) from groundnut (Arachis hypogaea) is the formation of resveratrol. The enzyme uses 4-coumaroyl-CoA, performs three condensations with malonyl-CoA, and folds the resulting tetraketide into a new aromatic ring system. We investigated the capacity for building novel and unusual polyketides from alternative substrates. Three types of products were obtained: (a) complete reaction (stilbene-type), (b) three condensations without formation of an aromatic ring (CTAL-type pyrone derailment), and (c) two condensations (BNY-type pyrone derailment). All product types were obtained from 4-fluorocinnamoyl-CoA and analogs in which the coumaroyl moiety was replaced by furan or thiophene. Only type (b) and (c) products were synthesized from other 4-substituted 4-coumaroyl-CoA analogs (-Cl, -Br, -OCH3). Benzoyl-CoA, phenylacetyl-CoA, and medium chain aliphatic CoA esters were poor substrates, and the majority of the products were of type (c). The results show that minor modifications can be used to direct the enzyme reaction to form a variety of different and new products. Manipulation of the biosynthesis of polyketides by synthetic analogs could lead to the development of a chemical library of pharmaceutically interesting novel polyketides. PMID- 11432744 TI - Oxidative folding of murine prion mPrP(23-231). AB - A systematic study of the oxidative folding of murine prion protein mPrP(23-231) is reported here. Folding of mPrP(23-231) involves formation of a single disulfide bond, Cys179-Cys214. Despite this simplicity, reduced mPrP(23-231) exhibits numerous unusual folding properties. In the absence of denaturant, folding of mPrP(23-231) is extremely sluggish, regardless of pH. The optimal pH for mPrP(23-231) folding was found to be 4-5. At pH 8.0, a condition that typically favors disulfide formation, folding of mPrP(23-231) hardly occurs, and it not facilitated by inclusion of redox agent. In the presence of denaturant (4 M urea or 2 M guanidine hydrochloride) and basic pH (8.0), reduced mPrP(23-231) refolds to the native structure quantitatively. The efficiency of folding can be further promoted by the presence of oxidized glutathione. At pH 4.0 and in the presence of 4 M urea, reduced mPrP(23-231) converts to three distinctive conformational isomers, unable to form the native structure. These unusual properties lead us to the following conclusions. The reduced mPrP(23-231) adopts a highly rigid structure with the two cysteines buried or situated apart. The presence of denaturant or low pH disrupts this rigid structure and lowers the energy barrier, which permits oxidation and refolding of the reduced mPrP(23 231). Under selected conditions, reduced mPrP(23-231) is capable of taking on multiple forms of stable conformational isomer that are segregated by energy barriers. PMID- 11432745 TI - The NADH-dependent reductase of a putative multicomponent tetrahydrofuran mono oxygenase contains a covalently bound FAD. AB - NADH-cytochrome c oxidoreductase activity specifically expressed during growth on tetrahydrofuran was detected in cell extracts of Pseudonocardia sp. strain K1. The enzyme catalyzing this reaction was purified to apparent homogeneity by a three-step purification procedure. It was characterized as a monomer of apparent molecular mass 40 kDa. Spectroscopic studies indicated that it contains an iron sulfur cluster and a flavin cofactor. An amount of 1 mol of flavin and 1 mol of iron was determined per mol of homogeneous protein. The N-terminal amino-acid sequence exhibited great similarity to the reductase component of various oxygenases. Cloning and sequencing of the corresponding gene designated as thmD revealed an ORF encoding a protein of 360 amino acids. An overall similarity of up to 38% was obtained to the NAD(P)H-acceptor reductase of several binuclear iron-containing mono-oxygenases. Conserved sequence motifs were identified that were similar to the chloroplast-type ferredoxin 2Fe-2S centre and to nucleotide binding domains. Studies on the flavin cofactor showed that it could not be removed from the protein by denaturation, indicating a covalent attachment. Spectroscopic studies revealed that the flavin is at the FAD level and covalently bound to the protein via the flavin 8alpha-methyl group. Thus, the isolated reductase component is the first enzyme of this type for which a covalent attachment of the flavin has been observed. PMID- 11432746 TI - Microperoxidase 8 catalyzed nitration of phenol by nitrogen dioxide radicals. AB - Microperoxidase 8 (MP8) is a heme octapeptide obtained by hydrolytic digestion of horse heart cytochrome c. At pH below 9, the heme iron is axially coordinated to the imidazole side chain of His18 and to a water molecule. Replacement of this weak ligand by H2O2 allows the formation of high-valent iron-oxo species which are responsible for both peroxidase-like and cytochrome P450-like activities of MP8. This paper shows that MP8 is able to catalyze the nitration of phenol by nitrite. The reaction requires H2O2 and is inhibited by ligands having a high affinity for the iron, catalase and radical scavengers. This suggests that the nitrating species could be NO2* radicals formed by the oxidation of nitrite by high-valent iron-oxo species. This new activity of MP8 opens a new access to nitro-aromatic compounds under mild conditions and validates the use of this minienzyme to mimick heme peroxidases, especially in the reactions of NO-derived species with biomolecules under oxidative stress conditions. PMID- 11432747 TI - Factor VII and single-chain plasminogen activator-activating protease: activation and autoactivation of the proenzyme. AB - Structural and biological characteristics of a recently described plasma serine protease, which displayed factor VII as well as pro-urokinase-activating properties in vitro, indicated a dual role for this factor VII-activating protease (FSAP) in hemostasis. Only the active protease (two-chain FSAP) has been isolated from plasma and from a prothrombin complex concentrate, whereas activators of the proenzyme have not been identified so far. After purification of the FSAP proenzyme from cryo-poor plasma by adsorption to an immobilized mAb and subsequent ion-exchange chromatography, activation to generate two-chain FSAP was followed by a direct chromogenic assay as well as by the ability of two-chain FSAP to activate pro-urokinase. Purified single-chain FSAP underwent autoactivation leading to the typical protease two-chain pattern and subsequent degradation products, as demonstrated by Western-blotting analysis using a site specific mAb. This autoactivation was significantly enhanced in the presence of heparin, whereas Ca2+ ions stabilized single-chain FSAP (the proenzyme) resulting in slower autoactivation kinetics. Correspondingly, the heparin-augmented reaction, which was associated with autodegradation particularly of the protease domain, was slowed down by co-incubation with Ca2+. Of the other proteases and cofactors tested, only urokinase (uPA) was able to generate the typical two-chain FSAP pattern. Studies with different forms of uPA suggest that the catalytic activity of pro-urokinase/uPA is needed to activate single-chain FSAP, indicating that it is the only hemostatic protease that can act as a physiological activator of FSAP. PMID- 11432748 TI - Regulation of laminin beta2 chain gene expression in human cancer cell lines. AB - The laminin beta2 chain is a basement membrane component expressed in a tissue- and developmental stage-specific manner. In this report we have examined the transcriptional and post-transcriptional regulation of the human laminin beta2 chain in human tumor cell lines. Both the A204 rhabdomyosarcoma and clone A colon carcinoma cells express the laminin beta2 chain mRNA, but only the A204 cells secrete laminin heterotrimers containing the beta2 chain. Segments of the beta2 chain gene promoter region were cloned into luciferase reporter vectors, and their ability to stimulate transcription was tested by transient transfection. Sequences downstream of the transcription start site between nucleotides +91 and +120 were found to be essential for luciferase activity in the two cell lines. Additional positive regulatory regions were present further upstream, between nucleotides -164 to -667 and between nucleotides -667 to -1724. Genomic DNA at the 3' end of the gene also appeared to have enhancer activity, as a 1.1-kb fragment located downstream of the last exon stimulated the luciferase activity of the nucleotides -667/+297 promoter segment approximately threefold. Alternative splicing of the first intron of the human laminin beta2 chain gene generates two isoforms of the 5' untranslated region of the beta2 chain mRNA. The translational efficiencies of the two laminin beta2 chain leaders did not differ significantly, when assayed by polysome profile analysis of endogenous clone A cell beta2 chain mRNA, transient transfection of chimeric beta2 chain leader/luciferase expression plasmids in clone A cells, and translation of in vitro synthesized RNAs in rabbit reticulocyte lysates. PMID- 11432749 TI - The variant tuf3 gene of Streptomyces coelicolor A3(2) encodes a real elongation factor Tu, as shown in a novel Streptomyces in vitro translation system. AB - In Streptomyces coelicolor, the regular and abundant elongation factor (EF)-Tu1 is encoded by tuf1, while the actual function of the highly divergent tuf3 gene product is not yet known. Expression of the latter could so far only be detected on the transcriptional level under stress conditions. In this paper we demonstrate the presence of low levels of EF-Tu3 in strains of the J1501 lineage. Enhanced expression was observed for J1501 glkA and relA deletion mutants, which lack glucose kinase and ribosome-bound ppGpp synthetase, respectively. To assess the putative translational capacities of EF-Tu3, a novel Streptomyces in vitro translation assay was designed, based on the full elimination by Ni2+ affinity adsorption of chromosomally encoded (His)6-tagged EF-Tu1 from a S. coelicolor cell-free extract. Translational activity of this system is totally dependent on the addition of purified EF-Tu species or on the presence of an additional elongation factor Tu in the extract, e.g. encoded by a plasmid-borne tuf gene. Using this EF-Tu-dependent translation system, we have established that S. coelicolor EF-Tu3 has translational capacities despite its striking deviation from the common prokaryotic EF-Tu sequence at positions involved in either aminoacyl-tRNA binding or interaction with the guanine-nucleotide exchange factor EF-Ts. PMID- 11432750 TI - A barley polyamine oxidase isoform with distinct structural features and subcellular localization. AB - Two cDNAs encoding polyamine oxidase (PAO) isoforms (BPAO1 and BPAO2) and the corresponding gene copies were isolated from barley cultivar Aura. Gene organization is not conserved between these two nonallelic coding sequences. Both precursor proteins include a cleavable N-terminal leader of 25 amino acids. N terminal sequencing of PAO purified from barley seedlings reveals a unique amino acid sequence corresponding to the BPAO2 N-terminus as predicted from the corresponding cDNA. BPAO2 has been purified, characterized and compared to maize PAO (MPAO), the best characterized member of this enzyme class. The two proteins show different pH optima for catalytic activity, Km and Vmax values with spermidine and spermine as substrates. Molecular modelling of BPAO2 reveals the same global fold as in MPAO. However, substitution of the active site residue Phe403 by a tyrosine, provides a rationale for the different catalytic properties of the two enzymes. In barley leaves PAO-specific activity is higher in isolated mesophyll protoplasts than in the extracellular fluids, whereas in maize the reverse is true. The C-terminus of BPAO2 shows homology with the endoplasmic reticulum retention signal that might be responsible for the subcellular localization observed. We conclude that BPAO2 is a symplastic PAO in barley mesophyll cells. Production of BPAO2 mRNA and the corresponding protein is induced by light, and has a different pattern of accumulation in leaves and coleoptiles. PMID- 11432751 TI - Characterization and mutagenesis of the recombinant N-acetylneuraminate lyase from Clostridium perfringens: insights into the reaction mechanism. AB - The N-acetylneuraminate lyase from Clostridium perfringens was expressed in Escherichia coli as a fusion protein with a His-tag and purified to homogeneity using metal chelate affinity and anion exchange chromatography. The purified enzyme has a pH optimum of 7.6 and a temperature optimum of 65-70 degrees C. In kinetic studies the lyase exhibits a Km of 3.2 mM for Neu5Ac and a Vmax of 27.5 U x mg(-1). To clarify the functional role of some putative active site residues, site-directed mutagenesis was performed. Lysine 161 was identified as the residue forming the Schiff base intermediate with the substrate. Tyrosine 133 was shown to be also a catalytically important residue; it seems to function as an acceptor for the proton of the C4 hydroxyl group, as already suggested by other groups. Furthermore, it is involved in stabilizing the Schiff base intermediate. Mutations of aspartate 187 and glutamate 188 indicate that both residues are involved in substrate binding. In this respect the carboxy group of aspartate 187 seems to be particularly important. Based on the results of these studies, a model of the reaction mechanism is discussed. PMID- 11432752 TI - Inactivation of Aeromonas hydrophila metallo-beta-lactamase by cephamycins and moxalactam. AB - Incubation of moxalactam and cefoxitin with the Aeromonas hydrophila metallo-beta lactamase CphA leads to enzyme-catalyzed hydrolysis of both compounds and to irreversible inactivation of the enzyme by the reaction products. As shown by electrospray mass spectrometry, the inactivation of CphA by cefoxitin and moxalactam is accompanied by the formation of stable adducts with mass increases of 445 and 111 Da, respectively. The single thiol group of the inactivated enzyme is no longer titrable, and dithiothreitol treatment of the complexes partially restores the catalytic activity. The mechanism of inactivation by moxalactam was studied in detail. Hydrolysis of moxalactam is followed by elimination of the 3' leaving group (5-mercapto-1-methyltetrazole), which forms a disulfide bond with the cysteine residue of CphA located in the active site. Interestingly, this reaction is catalyzed by cacodylate. PMID- 11432753 TI - The role of Asp42 in Escherichia coli inorganic pyrophosphatase functioning. AB - Excess of Mg2+ ions is known to inhibit the soluble inorganic pyrophosphatases (PPases). In contrast, the mutant Escherichia coli inorganic pyrophosphatase Asp42-->Asn is three times more active than native and retains its activity at high Mg2+ concentration. In this paper, another two mutant variants with Asp42 replaced by Ala or Glu were investigated to characterize the role of Asp42 in catalysis. pH-independent kinetic parameters of MgPPi hydrolysis and the dissociation constants for the activating and inhibitory Mg2+ ions were calculated. It was shown that Mg2+ inhibition of MgPPi hydrolysis by native PPase exhibited uncompetitive kinetics under the saturating substrate concentration. All three substitutions of Asp42 lead to a sharp decrease of inhibitory Mg2+ affinity to the enzyme. These findings allow determination of the sites of inhibitory and substrate Mg2+ ions binding to PPase. Common features of these mutants allow the conclusion that the function of Asp42 is to accurately coordinate the residues implicated in the substrate and the inhibitory Mg2+ ion binding to PPase active site. Structural analysis of PPase complexed with Mg2+ compared with PPase complexed with Mn2+ and reaction products confirms this supposition. PMID- 11432754 TI - Phosphorylation of seminal vesicle protein IV on Ser58 enhances its peroxidase stimulating activity. AB - In this study we show that SV-IV, a major immunomodulatory, anti-inflammatory, and sperm immunoprotective protein secreted from the rat seminal vesicle epithelium, acts in vitro as a substrate of protein kinase C (PKC) competing efficiently with H1 histone, a very well known PKC substrate. Electrospray mass spectrometry (ES-MS) analysis demonstrated that approximately 10% of the native SV-IV molecules were phosphorylated by PKC and that such a modification involved only a single serine residue (Ser58) out of the 22 occurring in the protein. Interestingly, this modification produced a substantial enhancement (approximately 50%) of the native SV-IV's ability to stimulate the activity of both horseradish peroxidase (POD) and selenium-dependent glutathione peroxidase (GPX), an enzyme that is known to protect the mammalian spermatozoa from oxidative stress and loss of motility in the female genital tract following ejaculation. In contrast, the phosphorylation of SV-IV on Ser58 did not produce any effect on the anti-inflammatory properties of SV-IV, as measured by its ability to inhibit the phospholipase A2. PMID- 11432755 TI - Peptide binding characteristics of the non-classical class Ib MHC molecule HLA-E assessed by a recombinant random peptide approach. AB - BACKGROUND: Increasing evidence suggests that the effect of HLA-E on Natural Killer (NK) cell activity can be affected by the nature of the peptides bound to this non-classical, MHC class Ib molecule. However, its reduced cell surface expression, and until recently, the lack of specific monoclonal antibodies hinder studying the peptide-binding specificity HLA-E. RESULTS: An in vitro refolding system was used to assess binding of recombinant HLA-E to either specific peptides or a nonamer random peptide library. Peptides eluted from HLA-E molecules refolded around the nonamer library were then used to determine a binding motif for HLA-E. Hydrophobic and non-charged amino acids were found to predominate along the peptide motif, with a leucine anchor at P9, but surprisingly there was no methionine preference at P2, as suggested by previous studies. CONCLUSIONS: Compared to the results obtained with rat classical class Ia MHC molecules, RT1-A1c and RT1-Au, HLA-E appears to refold around a random peptide library to reduced but detectable levels, suggesting that this molecule's specificity is tight but probably not as exquisite as has been previously suggested. This, and a previous report that it can associate with synthetic peptides carrying a viral sequence, suggests that HLA-E, similar to its mouse counterpart (Qa-1b), could possibly bind peptides different from MHC class I leader peptides and present them to T lymphocytes. PMID- 11432756 TI - Prophylactic cranial irradiation in small cell lung cancer: a systematic review of the literature with meta-analysis. AB - PURPOSE: A systematic review of the literature was carried out to determine the role of prophylactic cranial irradiation (PCI) in small cell lung cancer (SCLC). METHODS: To be eligible, full published trials needed to deal with SCLC and to have randomly assigned patients to receive PCI or not. Trials quality was assessed by two scores (Chalmers and ELCWP). RESULTS: Twelve randomised trials (1547 patients) were found to be eligible. Five evaluated the role of PCI in SCLC patients who had complete response (CR) after chemotherapy. Brain CT scan was done in the work-up in five studies and brain scintigraphy in six. Chalmers and ELCWP scores are well correlated (p < 0.001), with respective median scores of 32.6 and 38.8 %. This meta-analysis based on the available published data reveals a decrease of brain metastases incidence (hazard ratio (HR): 0.48; 95 % confidence interval (CI): 0.39 - 0.60) for all the studies and an improvement of survival (HR: 0.82; 95 % CI: 0.71 - 0.96) in patients in CR in favour of the PCI arm. Unfortunately, long-term neurotoxicity was not adequately described. CONCLUSIONS: PCI decreases brain metastases incidence and improves survival in CR SCLC patients but these effects were obtained in patients who had no systematic neuropsychological and brain imagery assessments. The long-term toxicity has not been prospectively evaluated. If PCI can be recommended in patients with SCLC and CR documented by a work-up including brain CT scan, data are lacking to generalise its use to any CR situations. PMID- 11432757 TI - A simple method to assess the oxidative susceptibility of low density lipoproteins. AB - BACKGROUND: Oxidative modification of low density lipoproteins (LDL) is recognized as one of the major processes involved in atherogenesis. The in vitro standardized measurement of LDL oxidative susceptibility could thus be of clinical significance. The aim of the present study was to establish a method which would allow the evaluation of oxidative susceptibility of LDL in the general clinical laboratory. RESULTS: LDL was isolated from human plasma by selective precipitation with amphipathic polymers. The ability of LDL to form peroxides was assessed by measuring thiobarbituric acid reactive substances (TBARS) after incubation with Cu2+ and H2O2. Reaction kinetics showed a three phase pattern (latency, propagation and decomposition phases) which allowed us to select 150 min as the time point to stop the incubation by cooling and EDTA addition. The mixture Cu2+/H2O2 yielded more lipoperoxides than each one on its own at the same time end-point. Induced peroxidation was measured in normal subjects and in type 2 diabetic patients. In the control group, results were 21.7 +/- 1.5 nmol MDA/mg LDL protein, while in the diabetic group results were significantly increased (39.0 +/- 3.0 nmol MDA/mg LDL protein; p < 0.001). CONCLUSION: a simple and useful method is presented for the routine determination of LDL susceptibility to peroxidation in a clinical laboratory. PMID- 11432758 TI - Effects of fat on carbohydrate absorption: more is not necessarily better. PMID- 11432759 TI - Influence of dietary fat on postprandial glucose metabolism (exogenous and endogenous) using intrinsically (13)C-enriched durum wheat. AB - The present study evaluates the influence of different amounts of fat added to starch on postprandial glucose metabolism (exogenous and endogenous). Nine women (24 (se 2) years old, BMI 20.4 (se 0.7) kg/m(2)) ingested 1 week apart 75 g glucose equivalent of (13)C-labelled starch in the form of pasta without (low fat; LF) or with 15 (medium fat; MF) or 40 (high fat; HF) g sunflower oil. During the 7 h following meal consumption, plasma glucose, non-esterified fatty acids, triacylglycerols (TG) and insulin concentrations, and endogenous (using [6,6 (2)H(2)]glucose) and exogenous glucose turnover were determined. With MF and HF meals, a lower postprandial glucose peak was observed, but with a secondary recovery. A decrease in exogenous glucose appearance explained lower glycaemia in HF. At 4 h after the HF meal the insulin, insulin:glucose and postprandial blood TG were higher than those measured after the LF and MF meals. Despite higher insulinaemia, total glucose disappearance was similar and endogenous glucose production was suppressed less than after the LF and MF meals, suggesting insulin resistance. Thus, the addition of a large amount of fat appears to be unfavourable to glucose metabolism because it leads to a feature of insulin resistance. On the contrary, the MF meal did not have these adverse effects, but it was able to decrease the initial glycaemic peak. PMID- 11432760 TI - Effects of dietary coconut oil on apolipoprotein B synthesis and VLDL secretion by calf liver slices. AB - Incorporation of coconut oil (CO) rich in lauric acid into the milk diet induces a lipid infiltration of the liver (steatosis) in 1-month-old calves. Among possible steps involved in diet-induced liver steatosis, the ability of the calf liver to synthesize apolipoprotein (Apo) B and to secrete it as part of VLDL particles was investigated. Liver samples were taken from calves fed for 17 d on a conventional milk replacer containing CO (n 5) and beef tallow (BT, n 4) as reference. Samples were cut into slices 0.5 mm thick and subsequently incubated for 12 h in a medium containing a [(35)S]methionine-[(35)S]cysteine mix and 0.8 mm-sodium laurate or oleate, the major fatty acids of CO and BT diets respectively. Concentrations of total [(35)S]proteins, [(35)S]albumin and [(35)S]ApoB in liver cells were 2-fold lower 0.0004 and 0.03 respectively) in CO- than in BT-fed calves. Although the total amount of proteins secreted (including albumin) was similar in both groups of calves, the amount of VLDL-[(35)S]Apo secreted was 2-fold lower (P = 0.004) in CO- than in BT-fed calves. These results suggest that a CO-enriched milk diet induces in preruminant calves a lipid infiltration of the liver by decreasing ApoB synthesis, leading to a reduction in secretion of VLDL particles. PMID- 11432761 TI - Feeding tuna oil to the sow at different times during pregnancy has different effects on piglet long-chain polyunsaturated fatty acid composition at birth and subsequent growth. AB - In an attempt to prevent decreases in piglet 20 : 4n-6 status at birth while increasing 22 : 6n-3 status, multiparous sows (eight per treatment) were allocated to one of three different treatments: a basal diet fed from day 63 of pregnancy to term; basal diet supplemented with tuna oil (17.5 g/kg) from day 63 to day 91 and then basal diet alone from day 92 to term; basal diet alone from day 63 to day 91 and then basal diet supplemented with tuna oil from day 92 to term. Tuna oil supplementation increased mainly 22 : 6n-3 intake. Supplementation with tuna oil between day 92 and term increased 22 : 6n-3 to a greater extent in all piglet tissues (brain, liver, retina and the remaining carcass) at birth than supplementation with tuna oil between days 63 and 91. However, while piglet 20 : 4n-6 decreased to a greater extent in liver and carcass when diets were supplemented with tuna oil between days 92 and term than between days 63 and 91, in the brain and retina, the reverse was true; 20 : 4n-6 was decreased to a greater extent between days 63 and 91 than between 92 and term. The effect of pregnancy nutrition on the growth of piglets until 7 d postweaning (35 d of age) was assessed after removing any residual effects of pregnancy treatment by cross fostering some piglets at birth. Piglets, the diets of whose dams had been supplemented with tuna oil during pregnancy, grew faster during the first 35 d of life than the progeny of sows fed only the basal diet. Feeding tuna oil to sows at different times during pregnancy therefore did not prevent decreases in piglet 20 : 4n-6 status at birth, but did suggest that changes in piglet brain 20 : 4n-6 status between days 63 and 91 of pregnancy were not reversible by later nutrition. Supplementing the diet of the pregnant sow with tuna oil had beneficial effects on postnatal piglet growth. PMID- 11432762 TI - High fat intake lowers hepatic fatty acid synthesis and raises fatty acid oxidation in aerobic muscle in Shetland ponies. AB - The metabolic effects of feeding soyabean oil instead of an isoenergetic amount of maize starch plus glucose were studied in ponies. Twelve adult Shetland ponies were given a control diet (15 g fat/kg DM) or a high-fat diet (118 g fat/kg DM) according to a parallel design. The diets were fed for 45 d. Plasma triacylglycerol (TAG) concentrations decreased by 55 % following fat supplementation. Fat feeding also reduced glycogen concentrations significantly by up to 65 % in masseter, gluteus and semitendinosus muscles (P < 0.05 and P < 0.01 and P < 0.01 respectively). The high-fat diet significantly increased the TAG content of semitendinosus muscle by 80 % (P < 0.05). Hepatic acetyl-CoA carboxylase and fatty acid synthase activities were 53 % (P < 0.01) and 56 % (P < 0.01) lower respectively in the high-fat group, but diacylglycerol acyltransferase activity was unaffected. Although carnitine palmitoyltransferase I (CPT-I) activity in liver mitochondria was not influenced, fat supplementation did render CPT-I less sensitive to inhibition by malonyl-CoA. There was no significant effect of diet on the activity of phosphofructokinase in the different muscles. The activity of citrate synthase was raised significantly (by 25 %; P < 0.05) in the masseter muscle of fat-fed ponies, as was CPT-I activity (by 46 %; P < 0.01). We conclude that fat feeding enhances both the transport of fatty acids through the mitochondrial inner membrane and the oxidative capacity of highly-aerobic muscles. The higher oxidative ability together with the depressed rate of de novo fatty acid synthesis in liver may contribute to the dietary fat-induced decrease in plasma TAG concentrations in equines. PMID- 11432763 TI - In vivo measurement of lipogenesis in ruminants using [1-(14)C]acetate. AB - A method for the measurement of the rate of lipogenesis in ruminants using a continuous intravenous infusion of [1-(14)C]acetate and measuring the rate of [1 (14)C]acetate incorporation into adipose tissue lipid was evaluated. Subcutaneous adipose tissue samples obtained by biopsy over the course of a 6 h continuous intravenous infusion of [1-(14)C]acetate into a wether and a steer maintained in a 'metabolic steady state' demonstrated that the incorporation of [1 (14)C]acetate into subcutaneous adipose tissue lipid was linear for the duration of the infusion period. Subsequent measures of rates of [1-(14)C]acetate incorporation into adipose tissue lipid were made on adipose tissue samples taken at a single time point during the infusion period. The technique was used to measure rates of lipogenesis in the subcutaneous adipose tissue of fourteen Hereford x Friesian steers that had been fed a pelleted diet of dried grass at a range of metabolizable energy (ME) intakes from 1.1 x ME requirement for maintenance to ad libitum for 11 weeks. Rates of lipogenesis increased linearly with increasing ME intake. It was concluded that the method is an effective technique for measuring rates of lipogenesis in specific adipose tissue depots in vivo in ruminants. PMID- 11432764 TI - Evaluation of a model for total body protein mass based on dual-energy X-ray absorptiometry: comparison with a reference four-component model. AB - The aim of the present study was to evaluate a model of body composition for assessing total body protein (TBP) mass using dual-energy X-ray absorptiometry (DXA), with either measured or assumed total body water (TBW); it was intended to provide a less complex or demanding alternative technique to, for example, the four-component model (4-CM). The following measurements were obtained in healthy adults (n 46) aged 18--62 years, and children (n 30) aged 8--12 years: body weight (BWt), body volume (BV; under-water weighing), TBW ((2)H-dilution space or predicted using an assumed hydration fraction of fat-free mass (HF(ffm))), bone mineral content (BMC; DXA) and fat-free soft tissue (FFST; DXA). TBP was calculated using the 4-CM (TBP = 3.05BWt -- 0.290TBW -- 2.734BMC -- 2.74BV) and the DXA model (TBP = FFST -- 0.2302BMC -- TBW). DXA measurements were obtained using the Lunar DPX (Lunar Radiation Corporation, Madison, WI, USA) or Hologic QDR 1000/W (Hologic, Waltham, MA, USA). Precision of the DXA model for TBP with measured TBW (4.6--6.8 % mean TBP) was slightly worse than the 4-CM (4.0--5.4 %), whereas that modelled with assumed HF(ffm) was more precise (2.4--5.2 %) because it obviated imprecision associated with measuring TBW. Agreement between the 4-CM and DXA model with measured TBW was also worse (e.g. bias, 15 % of the mean; 95 % limits of agreement up to +/-39 % for adults measured on the Lunar DPX) than when a constant for HF(ffm) was assumed (3.7 % and +/-21 % respectively). Most of the variability in agreement between these various models was due to interpretation of biological factors, rather than to measurement imprecision. Therefore, the DXA model, which is less complex and demanding than the 4-CM, is of value for assessing TBP in groups of healthy subjects, but is of less value for individuals in whom there may be substantial differences from reference 4-CM estimates. PMID- 11432765 TI - The effect of fasting at different ages on growth and tissue dynamics in the small intestine of the young chick. AB - The small intestines of hatching chicks undergo rapid developmental changes in the immediate post-hatch period when the birds are making the transition from endogenous nutrient supply from yolk to dependence on exogenous feed. This transition usually only begins 48 h or more after hatching, owing to logistical considerations of production. The effects of fasting for 48 h at different times during this critical period on small intestinal development and enterocyte dynamics were examined by morphometric determinations and use of staining for proliferative-cell nuclear antigen and 5-bromo-2-deoxyuridine. The effects of fasting were specific to both time of fasting and the intestinal segment examined. Decreased development was found in the duodenum and jejunum, but was less apparent in the ileum. Fasting between 0 and 48 h decreased crypt size in the duodenum and jejunum, the number of crypts per villus, crypt proliferation, villus area and the rate of enterocyte migration. Fasting at later times resulted in smaller effects, although the jejunum appeared to be the most sensitive of the intestinal segments. Growth was correlated with the number of cells in the crypts, the number of cells along the villus and the segment surface area. The common practice whereby feed is first available to chicks more than 48 h post hatch may depress subsequent development. PMID- 11432766 TI - Effect of high temperature on feeding behaviour and heat production in group housed young pigs. AB - To assess the acclimation of pigs to heat stress, the effects of high (33 degrees C) or thermoneutral (23 degrees C) constant temperatures on feeding behaviour and components of energy balance were studied in group-housed young pigs. Three groups of five pigs were used at each temperature. After 1 week of adaptation, voluntary feed intake (VFI) and heat production (HP) were recorded for thirteen consecutive days. Animals were fed ad libitum. Fasting HP was measured on the last day. Average initial body weights (BW) were 21.4 and 20.9 kg at 23 and 33 degrees C respectively. Feeding behaviour was measured individually and rate of feed intake and characteristics of feeding behaviour were calculated. The O(2) consumption, CO(2) production and physical activity of the group were used to calculate total HP (HP(tot)) and its components, i.e. fasting HP (HP(fas)), HP due to physical activity (HP(act)) and thermic effect of feed (TEF). The BW gain and VFI were reduced by 37 and 30 % respectively at 33 degrees C. The decrease in VFI corresponded to reduced consumption time (-34 %) and size of the meals (-32 %). Feeding behaviour was mostly diurnal (66 % of the VFI), and the rate of feed intake (28 g/min) was not affected by temperature. Daily HP(tot), HP(fas) and TEF, expressed per kg metabolic weight (BW(0.60)), were significantly decreased at 33 degrees C by 22, 18 and 35 % respectively, whereas HP(act) was not affected; TEF expressed per g feed was not affected (2 kJ/g). The decrease in HP(tot) at 33 degrees C was caused by a reduction in TEF and HP(fas) (kJ/d per/kg BW(0.60)), which are both related to reduction in VFI. PMID- 11432767 TI - Are changes in food consumption patterns associated with lower biochemical zinc status among women from Dunedin, New Zealand? AB - Reductions in red meat and increases in cereals in the diet may compromise the intake and bioavailability of Zn. In this cross-sectional study of 330 premenopausal New Zealand women aged 18--40 years, we have assessed the inter relationships among dietary intakes (via computer-administered food-frequency questionnaire), biochemical Zn status, and anthropometric indices, and compared our results with earlier data. Fasting serum (12.00 (sd 1.36) micromol/l) and hair Zn (2.71 (sd 0.36) micromol/g) were lower than those for young Dunedin, New Zealand, women in 1973 (non-fasting serum Zn 18.6 (sd 4.6) micromol/l, hair Zn 2.99 (sd 0.35) micromol/g). Further, our mean serum Zn was at the 25th percentile of the US National Health and Nutrition Examination Survey (NHANES) (1976--1980) reference sample for women aged 20--44 years. Meat-poultry-fish contributed only 28 % total Zn in the present study, a level comparable with that from cereals nuts-legumes (27 %), compared to about 40 % in 1989. Significant negative correlations existed between serum Zn and dietary [phytate]:[Zn] molar ratios (r 0.163, 35 % had diets with [phytate]:[Zn] >15, a level said to compromise Zn status. Mean serum Zn of a subgroup of non-oral contraceptive users free of infection was higher in the red-meat eaters (n 149) compared with non-red-meat eaters (n 48) (12.2 v. 11.8 micromol/g, In contrast, serum Zn was lower in those with dietary [phytate]:[Zn] ratios >15 v. <15 (i.e. 11.9 v. 12.3 micromol/l, We postulate that the lower biochemical Zn status of these New Zealand women may be associated in part with changes in food selection patterns, which have led to a reduction in the bioavailability of dietary Zn. PMID- 11432768 TI - Inter-pregnancy folate and iron status of women in an inner-city population. AB - The purpose of the present study was to evaluate whether micronutrient supplementation improved the nutritional status of women with poor diets during the inter-pregnancy interval. Fifty-five women who had given birth to a low birth weight baby (<2.5 kg), and who planned to have a further pregnancy, were recruited to a prospective randomised study in East London, UK. Of the fifty-five mothers recruited, forty-four (78 %) met fewer than four of sixteen dietary reference values according to the information provided in a 7 d diet diary, and were categorised as having an 'inadequate' diet. Half of the mothers in the 'inadequate'-diet group were randomly assigned to receive a micronutrient and a single cell oil supplement containing docosahexaenoic acid. All participants received dietary advice based on analysis of their diet diaries, and general lifestyle advice on preparing for pregnancy. Mothers had a blood sample taken at 3 and 9 months post-partum to measure their folate, Fe stores and fatty acid status. Mean serum and erythrocyte folate levels increased significantly between 3 and 9 months post-partum in both the adequate-diet group and the supplemented group. At 9 months post-partum, over half of the unsupplemented, inadequate-diet group remained severely deficient in folate (serum folate <230 nmol/l) and had low serum ferritin levels (<15 microg/l). The high prevalence of inadequate diets in this inner-city population and the low motivation of women to participate in a nutrition programme suggests that consideration should be given to the provision of free folate and Fe supplements to all women in this and similar populations, or at least to women who have delivered a low birth weight baby, who plan further pregnancies. PMID- 11432769 TI - Effects of whole wheat dilution v. substitution on coccidiosis in broiler chickens. AB - To test the hypothesis that inclusion of whole cereals in the diet of broiler chickens reduces the severity of a coccidial infection, the effects were investigated, in birds infected with Eimeria acervulina, of feeding a complete pelleted control food, the control food diluted with whole wheat (400 g/kg), or a diet in which ground wheat in the pellets was substituted with whole wheat (400 g/kg) so as to achieve the same composition as the control diet. In the weeks prior to and after infection, (days 14--20 and days 21--27), no significant differences in performance were observed between birds fed the complete pellet and substituted feeds. Birds fed the diluted feeds had significantly lower gains per unit of feed and numerically lower weight gains than the birds fed the control and substituted feeds during this period. No significant differences in performance were observed between treatments from days 28--34. Diluting feeds with whole wheat (400 g/kg) significantly increased ileal digesta viscosity levels, compared with feeding the control and substituted feeds in birds dissected on day 21, although no significant differences in digesta viscosity levels were observed in birds dissected on day 35. Whole wheat feeding, either by dilution or substitution, significantly increased gizzard sizes in birds dissected on day 21 and day 35. Neither dilution nor substitution of feeds with whole wheat (400 g/kg) significantly affected the level of an Eimeria acervulina infection, as measured by daily and total faecal oocyst yields. PMID- 11432770 TI - Cardiovascular actions of chicken-meat extract in normo- and hypertensive rats. AB - The cardiovascular actions of a commercial chicken-meat extract known as Brand's Essence of Chicken (Cerebos Pacific Ltd, Singapore; BEC) were investigated in normo- and hypertensive rats. The spontaneously-hypertensive rat (SHR), Wistar Kyoto rat (WKY) and Sprague Dawley rat (SD) were used. The effect of oral feeding of BEC on hypertension, cardiac hypertrophy and arteriosclerosis in these animals was studied. The data showed the following effects of oral feeding of BEC: (1) feeding for 30 d did not affect the blood pressure and heart rate (determined telemetrically) of adult SHR and WKY; (2) feeding for 90 d did not affect the development of hypertension in 1-month-old prehypertensive SHR; (3) feeding for 4 d dose-dependently (0.2--3.2 ml/kg per d) attenuated cardiac hypertrophy in experimentally-induced (coarctation of the abdominal aorta) cardiac hypertrophic SD; (4) feeding to 1-month-old prehypertensive SHR for 11 months did not affect the age-related development of hypertension in this animal; (5) there was significant attenuation of the age-related development of hypertension (determined by tail-cuff plethysmography) in the WKY (P = 0.011) when the animals drank an average of 7.5 ml BEC/kg body weight per d, measured during the last 2 months of the 11-month treatment period; (6) there was chronic, as in the previous treatment, attenuation of the age-related development of cardiac hypertrophy and arteriosclerosis (quantified morphometrically) in the SHR when the animals drank an average of 2.4 ml BEC/kg per d, measured during the last 2 months of the 11-month treatment period. A parallel study using laboratory prepared chicken-meat and pork extracts showed that the former, but not the latter, attenuated cardiac hypertrophy in experimentally-induced cardiac hypertrophic SD. These findings, showing that chicken-meat extract (both BEC and laboratory prepared) could have anti-cardiac hypertrophic, anti-hypertensive and anti-arteriosclerotic actions, were unexpected and provoking, and would challenge nutritional scientists with an interest in meat consumption and cardiovascular diseases. PMID- 11432771 TI - BMI does not accurately predict overweight in Asian Indians in northern India. AB - Asian Indians are at high risk for the development of atherosclerosis and related complications, possibly initiated by higher body fat (BF). The present study attempted to establish appropriate cut-off levels of the BMI for defining overweight, considering percentage BF in healthy Asian Indians in northern India as the standard. A total of 123 healthy volunteers (eighty-six males aged 18--75 years and thirty-seven females aged 20--69 years) participated in the study. Clinical examination and anthropometric measurements were performed, and percentage BF was calculated. BMI for males was 21.4 (sd 3.7) kg/m(2) and for females was 23.3 (sd 5.5) kg/m(2). Percentage BF was 21.3 (sd 7.6) in males and 35.4 (sd 5.0) in females. A comparison of BF data among Caucasians, Blacks, Polynesians and Asian ethnic groups (e.g. immigrant Chinese) revealed conspicuous differences. Receiver operating characteristic (ROC) curve analysis showed a low sensitivity and negative predictive value of the conventional cut-off value of the BMI (25 kg/m(2)) in identifying subjects with overweight as compared to the cut-off value based on percentage BF (males >25, females >30). This observation is particularly obvious in females, resulting in substantial misclassification. Based on the ROC curve, a lower cut-off value of the BMI (21.5 kg/m(2) for males and 19.0 kg/m(2) for females) displayed the optimal sensitivity and specificity, and less misclassification in identification of subjects with high percentage BF. Furthermore, a novel obesity variable, BF:BMI, was tested and should prove useful for interethnic comparison of body composition. In the northern Indian population, the conventional cut-off level of the BMI underestimates overweight and obesity when percentage BF is used as the standard to define overweight. These preliminary findings, if confirmed in a larger number of subjects and with the use of instruments having a higher accuracy of BF assessment, would be crucial for planning and the prevention and treatment of various obesity-related metabolic diseases in the Asian Indian population. PMID- 11432772 TI - Molecular mechanisms of the decision between life and death: regulation of apoptosis by apoptosis signal-regulating kinase 1. AB - Coordination and balance between cell survival and apoptosis is crucial for normal development and homeostasis of multicellular organisms. Defects in control of this balance may contribute to a variety of diseases including cancer, autoimmune and neurodegenerative conditions. Although a large number of pro- and anti-apoptotic factors acting for or against the final death event have been and are being discovered at an extraordinary pace with the recent progress in this area, the molecular mechanisms determining whether a cell lives or dies are not fully understood. Phosphorylation and dephosphorylation of intracellular effector molecules are the most common and important regulatory mechanisms in signal transduction and control a variety of cellular events from cell growth to apoptosis. Apoptosis signal-regulating kinase 1 (ASK1) is a member of the mitogen activated protein (MAP) kinase kinase kinase family, which activates both the SEK1-JNK and MKK3/6-p38 MAP kinase pathways and constitutes a pivotal signaling pathway in cytokine- and stress-induced apoptosis. This review provides recent findings on the molecular mechanisms which determine cell fate such as survival, proliferation, differentiation or apoptosis, with special focus on the regulatory mechanisms of ASK1-mediated apoptosis. PMID- 11432773 TI - Crystallographic studies on damaged DNAs: III. N(4)-methoxycytosine can form both Watson-Crick type and wobbled base pairs in a B-form duplex. AB - To investigate the mutation mechanism of purine transition in DNA damaged with methoxyamine, a DNA dodecamer with the sequence d(CGCGAATTmo(4)CGCG), where mo(4)C is 2'-deoxy-N(4)-methoxycytidine, has been synthesized and its crystal structure determined. Two dodecamers form a B-form duplex. Electron density maps clearly show that one of the two mo(4)C residues forms a pair with a guanine residue of the opposite strand, the geometry being the canonical Watson-Crick type, and that the other mo(4)C residue forms a wobble pair with the opposite guanine residue. These two pairings are ascribed to the tautomerization of the methoxylated cytosine moieties between the amino and imino forms. PMID- 11432774 TI - On the effect of sodium dodecyl sulfate on the structure of beta-galactosidase from Escherichia coli. A fluorescence study. AB - An understanding of the structure-function relationship of proteins under different chemical-physical conditions is of fundamental importance for an understanding of their structure and function in cells. In this paper we report the effects of sodium dodecyl sulfate and temperature on the structure of beta galactosidase from Escherichia coli, as monitored by fluorescence spectroscopy. The structure of the protein was studied in the temperature range of 10-60 degrees C in the absence and presence of sodium dodecyl sulfate by frequency domain measurement of the intrinsic fluorescence intensity and anisotropy decays. The time-resolved fluorescence data in the absence of SDS indicated that at 10 degrees C the tryptophanyl emission decays were well described by a three exponential decays model, and that the temperature increase resulted in shortening of the long-lived component with little change in the short- and middle-lived components. The addition of SDS to the protein solution also affected the long-lived component. The effects of the detergent and temperature on the enzyme structure were also investigated by means of quenching experiments and anisotropy decays. The obtained results showed that the presence of SDS confers more flexibility to the protein structure, and suggest a strict relation between enzyme activity and protein flexibility. PMID- 11432775 TI - An economical method for (15)N/(13)C isotopic labeling of proteins expressed in Pichia pastoris. AB - We report a new and cost-effective approach to prepare (15)N/(13)C labeled proteins for NMR using the Pichia pastoris expression system. Four protocols (P1 to P4) were defined and compared using recombinant Ovine interferon-tau (rOvIFN tau). Our results demonstrate that in order to get full incorporation of (15)N and (13)C, the isotopes are not totally required during the initial growth phase of P. pastoris culture. The addition of small amounts of (15)N and (13)C compounds 6 h prior to the methanol induction phase is sufficient to obtain 99% incorporation of heavy isotopes into the protein. Our optimized protocol P4 is two-thirds less costly than the classical method using (15)N and (13)C isotopes during the entire growth phase. PMID- 11432776 TI - PKNbeta interacts with the SH3 domains of Graf and a novel Graf related protein, Graf2, which are GTPase activating proteins for Rho family. AB - PKNbeta is a novel isoform of PKNalpha, which is one of the target protein kinases for the small GTPase Rho. By yeast two-hybrid screening of a human embryonic kidney 293 cell cDNA library with the PKNbeta linker region containing proline-rich motifs as a bait, clones encoding Graf (GAP for Rho Associated with Focal adhesion kinase) and a novel Graf-related protein, termed Graf2, were isolated. The full length of Graf2 contains a putative PH domain, a RhoGAP domain, and an SH3 domain as well as Graf. Northern and Western blot analyses demonstrated that Graf2 is expressed in several tissues, with the highest expression in skeletal muscle. Recombinant Graf2 exhibited GTPase-activating activity toward the small GTPase RhoA and Cdc42Hs, but not toward Rac1, in vitro. The SH3 domains of Graf and Graf2 purified from Escherichia coli bound directly to PKNbeta. Graf or Graf2 was co-immunoprecipitated with PKNbeta in COS-7 cells transiently transfected with Graf or Graf2 and PKNbeta expression constructs. The catalytically active form of PKNbeta phosphorylated Graf and Graf2 in vitro. The interplay of PKNbeta and the GTPase-activating proteins, Graf and Graf2, may offer a novel mechanism regulating the Rho-mediated signaling. PMID- 11432777 TI - Correlative fine specificity of several Thomsen-Friedenreich disaccharide-binding proteins with an effect on tumor cell proliferation. AB - Epithelial cancer cells show increased cell surface expression of mucin antigens with aberrant O-glycosylation, notably type I core (Galbeta1-3GalNAcalpha), termed Thomsen-Friedenreich disaccharide (TFD), a chemically well-defined carbohydrate antigen with a proven link to malignancy. Several TFD-binding proteins influence the proliferation of cells to which they bind. We studied the fine specificity of TFD-binding proteins and its relationship with epithelial tumor cell proliferation. Competitive binding assays against asialoglycophorin showed that Agaricus bisporus lectin (ABL) and human anti-TFD monoclonal antibody (mAb) TF1 were inhibited only by TFD and its alpha-derivatives. Peanut agglutinin (PNA), mAb TF2, and mAb TF5 were also inhibited by other carbohydrates such as lacto-N-biose (Galbeta1-3GlcNAc), lactose, and (Mealpha or beta) Gal, indicating lower recognition of the axial C-4 hydroxyl group position of GalNAc from TFD, and the major relevance of the terminal Gal on interaction of these three TFD binding proteins. In the direct glycolipid-binding assay, ABL bound mostly to alpha-anomeric TFD-bearing glycolipids, whereas PNA interacted mainly with beta linked TFD. Of the three anti-TFD mAbs analyzed, all bound N5b (terminal beta TFD), but only TF2 interacted with N6 (terminal alpha-TFD). These findings indicate that TFD-binding proteins that stimulate the proliferation of epithelial tumor cell lines recognize mainly a terminal beta-Gal region of beta-linked TFD, whereas ABL, which inhibits the proliferation of these tumor cells, binds mainly to subterminal GalNAc of alpha-anomeric TFD. PMID- 11432778 TI - Expression and exon/intron organization of two medaka fish homologs of the mammalian guanylyl cyclase A. AB - Two cDNA clones (OlGC2 and OlGC7) and their genomic DNA clones encoding medaka fish homologs of mammalian natriuretic peptide receptor/membrane guanylyl cyclase A (GC-A) were isolated, and their complete nucleotide sequences were determined. The open reading frame predicts a protein of 1,063 amino acids for OlGC2 cDNA (4,283 bp), and one of 1,055 amino acids for OlGC7 cDNA (3,721 bp), respectively. Northern blot analyses demonstrated 4.7 kb OlGC2 transcripts in the kidney and gill, and 4.0 kb OlGC7 transcripts in the kidney, brain, and ovary, while RNase protection analyses revealed that both genes are expressed in various adult organs. Both the OlGC2 (about 33.0 kbp) and OlGC7 (about 44.3 kbp) genes consist of 22 exons with an exon/intron organization similar to those of the human GC-A gene (about 16.6 kbp) and medaka fish GC-B homolog gene (OlGC1, about 93 kbp). Intron 4 of OlGC2 contains two repeated sequence (RS) clusters, designated as RS1 (about 1 kbp) and RS2 (about 5 kbp), consisting of nucleotide 5' AGCCTCTGCTCCTCCTTC-3'. In addition, many identical but variably sized nucleotide sequences were found in introns in OlGC1, OlGC2, OlGC6, and OlGC7. The OlGC2 and OlGC7 genes both have no apparent TATA box in the 5' flanking region upstream of the putative transcription initiation point, but several consensus sequences for cis-regulatory elements, including C/EBP, CREB, NF-IL6, and Sp1 and AP-2, NF-IL6, c-Myb, and Sp1 are present in the 5'-flanking region of OlGC2 and OlGC7, respectively. PMID- 11432779 TI - Amino acids in the N-terminal region regulate the photocycle of photoactive yellow protein. AB - The spectroscopic properties of photoactive yellow protein (PYP) partially digested by chymotrypsin were studied. Chymotrypsin yielded three major products that were yellow but distinguishable by SDS-PAGE. They were readily separated by DEAE-Sepharose column chromatography. Protein sequencing and mass spectrometry demonstrated that chymotrypsin cleaved the N-terminal 6, 15, or 23 amino acids (T6, T15, and T23). The blue-shifts of the absorption maxima and the increases in the apparent pK(a) of the chromophores relative to those of intact PYP were less than 4 nm and 0.2, respectively. The absorption spectra of the near-UV intermediates produced from T6, T15, and T23 were identical to that of intact PYP, but with lifetimes that were 140, 2,300, and 4,500 times longer, respectively. These observations suggest that the recovery of the dark state of PYP from the near-UV intermediate is accelerated by the N-terminal region, and that this region acts as a regulatory factor for the photocycle of PYP. PMID- 11432780 TI - Detection of the protein-protein interaction between cyclic AMP receptor protein and RNA polymerase, by (13)C-carbonyl NMR. AB - Cyclic AMP receptor protein (CRP) plays a key role in the transcription regulation of many prokaryotic genes. Upon the binding of cyclic AMP, CRP is allosterically activated, binds to target DNA sites, and interacts with RNA polymerase. Although the protein-protein interaction between CRP and RNA polymerase is known to be important for the transcription initiation of the target genes, its structural understanding is still lacking, particularly due to the high molecular mass (approximately 120 kDa) of the protein complex. We assigned all of the (13)C-carbonyl resonances of methionine residues in CRP by using the double labeling and the enzyme digestion techniques. The result of (13)C-carbonyl NMR experiment on [(13)C'-Met]-CRP in the presence of both cyclic AMP and RNA polymerase alpha subunit showed that the two proteins interact with each other in solution in the absence of DNA via the region around the residues from Met 157 to Met 163 in CRP. The results also showed the effectiveness of the selective labeling and (13)C-carbonyl NMR spectroscopy in the specific detection of the protein-protein interaction between large molecules. PMID- 11432781 TI - Genetic analysis of the isc operon in Escherichia coli involved in the biogenesis of cellular iron-sulfur proteins. AB - The iron-sulfur (Fe-S) cluster, the nonheme-iron cofactor essential for the activity of many proteins, is incorporated into target proteins with the aid of complex machinery. In bacteria, several proteins encoded by the iscRSUA-hscBA-fdx ORF3 cluster (isc operon) have been proposed to execute crucial tasks in the assembly of Fe-S clusters. To elucidate the in vivo function, we have undertaken a systematic mutational analysis of the genes in the Escherichia coli isc operon. In all functional tests, i.e. growth rate, nutritional requirements and activities of Fe-S enzymes, the inactivation of the iscS gene elicited the most drastic alteration. Strains with mutations in the iscU, hscB, hscA, and fdx genes also exhibited conspicuous phenotypical consequences almost identical to one another. The effect of the inactivation of iscA was small but appreciable on Fe-S enzymes. In contrast, mutants with inactivated iscR or ORF3 showed virtually no differences from wild-type cells. The requirement of iscSUA-hscBA-fdx for the assembly of Fe-S clusters was further confirmed by complementation experiments using a mutant strain in which the entire isc operon was deleted. Our findings support the conclusion that IscS, via cysteine desulfurase activity, provides the sulfur that is subsequently incorporated into Fe-S clusters by assembler machinery comprising of the iscUA-hscBA-fdx gene products. The results presented here indicate crucial roles for IscU, HscB, HscA, and Fdx as central components of the assembler machinery and also provide evidence for interactions among them. PMID- 11432782 TI - Association of phosphatidylinositol 3-kinase composed of p110beta-catalytic and p85-regulatory subunits with the small GTPase Rab5. AB - A family of phosphatidylinositol 3-kinases (PI 3-kinase), comprising three major classes (I-III) in terms of substrate specificity and regulation, play important roles in a variety of cell functions. We previously reported that the class-I heterodimeric PI 3-kinase consisting of p110beta-catalytic and p85-regulatory subunits is synergistically activated by two different types of membrane receptors, one possessing tyrosine kinase activity and the other activating trimeric G proteins. Here we report an additional unique feature of the p110beta/p85 PI 3-kinase. The small GTPase Rab5 was identified as a binding protein for the p110beta-catalytic subunit in a yeast two-hybrid screening system. The interaction appears to require at least two separated amino-acid sequences present specifically in the beta isoform of p110 and the GTP-bound form of Rab5. The expressions of constitutively active and dominant negative mutants of Rab5 in THP-1 cells induce the stimulation and inhibition, respectively, of protein kinase B activity, which is dependent on the PI 3-kinase product phosphatidylinositol 3,4,5-triphosphate. These results suggest that there is a specific interaction between GTP-bound Rab5 and the p110beta/p85 PI 3-kinase, leading to efficient coupling of the lipid kinase product to its downstream target, protein kinase B. PMID- 11432783 TI - Regulatory mechanism of human connective tissue growth factor (CTGF/Hcs24) gene expression in a human chondrocytic cell line, HCS-2/8. AB - CTGF/Hcs24 is a multi-functional growth factor that potentiates either the growth or differentiation of mesenchymal cells, according to the biological conditions. Among various functional aspects of CTGF/Hcs24, it is especially notable that CTGF/Hcs24 may promote endochondral ossification in growth cartilage through all stages, and it is highly expressed in a human chondrosarcoma-derived chondrocytic cell line (HCS-2/8). In this study, to clarify the regulatory mechanism of CTGF/Hcs24 gene expression in chondrocytes, we analyzed the transcriptional activity of the CTGF/Hcs24 promoter and the effect of the CTGF/Hcs24 3' untranslated region (3'-UTR) on gene expression in HCS-2/8 by means of an established DNA transfection and luciferase reporter gene assay system. As a result, the luciferase activity of the CTGF/Hcs24 promoter was found to be remarkably high in HCS-2/8. The 3'-UTR of the CTGF/Hcs24 gene strongly repressed the luciferase activity in HCS-2/8, when it was linked to the downstream of the luciferase reporter gene, suggesting its functionality also in chondrocytic cells. Deletion analysis of the CTGF/Hcs24 promoter clarified a major segment responsible for the enhanced CTGF/Hcs24 promoter activity in HCS-2/8. The TGF beta response element in the DNA segment was active in HCS-2/8, and point mutations in the element moderately decreased the highly maintained promoter activity with total loss of TGF-beta responsiveness. These results indicate that the strong expression of the CTGF/Hcs24 gene in HCS-2/8 was mainly caused by high transcriptional activity of the CTGF/Hcs24 promoter, and that the TGF-beta response element is one of the critical elements that support the high transcription activity. PMID- 11432784 TI - Substrate recognition mechanism of thermophilic dual-substrate enzyme. AB - Aspartate aminotransferase from an extremely thermophilic bacterium, Thermus thermophilus HB8 (ttAspAT), has been believed to be specific for an acidic substrate. However, stepwise introduction of mutations in the active-site residues finally changed its substrate specificity to that of a dual-substrate enzyme. The final mutant, [S15D, T17V, K109S, S292R] ttAspAT, is active toward both acidic and hydrophobic substrates. During the course of stepwise mutation, the activities toward acidic and hydrophobic substrates changed independently. The introduction of a mobile Arg292* residue into ttAspAT was the key step in the change to a "dual-substrate" enzyme. The substrate recognition mechanism of this thermostable "dual-substrate" enzyme was confirmed by X-ray crystallography. This work together with previous studies on various enzymes suggest that this unique "dual-substrate recognition" mechanism is a feature of not only aminotransferases but also other enzymes. PMID- 11432785 TI - Improved autoprocessing efficiency of mutant subtilisins E with altered specificity by engineering of the pro-region. AB - Modification of substrate specificity of an autoprocessing enzyme is accompanied by a risk of significant failure of self-cleavage of the pro-region essential for activation. Therefore, to enhance processing, we engineered the pro-region of mutant subtilisins E of Bacillus subtilis with altered substrate specificity. A high-activity mutant subtilisin E with Ile31Leu replacement (I31L) as well as the wild-type enzyme show poor recognition of acid residues as the P1 substrate. To increase the P1 substrate preference for acid residues, Glu156Gln and Gly166Lys/Arg substitutions were introduced into the I31L gene based upon a report on subtilisin BPN' [Wells et al. (1987) Proc. Natl. Acad. Sci. USA 84, 1219-1223]. The apparent P1 specificity of four mutants (E156Q/G166K, E156Q/G166R, G166K, and G166R) was extended to acid residues, but the halo forming activity of Escherichia coli expressing the mutant genes on skim milk containing plates was significantly decreased due to the lower autoprocessing efficiency. A marked increase in active enzyme production occurred when Tyr(-1) in the pro-region of these mutants was then replaced by Asp or Glu. Five mutants with Glu(-2)Ala/Val/Gly or Tyr(-1)Cys/Ser substitution showing enhanced halo forming activity were further isolated by PCR random mutagenesis in the pro region of the E156Q/G166K mutant. These results indicated that introduction of an optimum arrangement at the cleavage site in the pro-region is an effective method for obtaining a higher yield of active enzymes. PMID- 11432786 TI - X-ray crystalline structures of pyrrolidone carboxyl peptidase from a hyperthermophile, Pyrococcus furiosus, and its cys-free mutant. AB - In order to elucidate the mechanism of the thermostability of proteins from hyperthermophiles, X-ray crystalline structures of pyrrolidone carboxyl peptidase from a hyperthermophile, Pyrococcus furiosus (PfPCP), and its mutant protein with Ser substituted at Cys142 and Cys188 were determined at 2.2 and 2.7 A resolution, respectively. The obtained structures were compared with those previously reported for pyrrolidone carboxyl peptidases from a hyperthermophilie, Thermococcus litoralis (TlPCP), and from a mesophile, Bacillus amyloliquefaciens (BaPCP). The PfPCP structure is a tetramer of four identical subunits similar to that of the TlPCP and BaPCP. The largest structural changes among the three PCPs were detected in the C-terminal protrusion, which interacts with that of another subunit. A comparison of the three structures indicated that the high stability of PfPCP is caused by increases in hydrophobic interactions and hydrogen bonds, the formation of an intersubunit ion-pair network, and improvement to an ideal conformation. On the basis of the structures of the three proteins, it can be concluded that PfPCP does not have any special factors responsible for its extremely high stability and that the conformational structure of PfPCP is superior in its combination of positive and negative stabilizing factors compared with BaPCP. PMID- 11432787 TI - A novel alpha-amino-acid esterase from Bacillus mycoides capable of forming peptides of DD- and DL-configurations. AB - A novel alpha-amino-acid esterase possessing some properties favorable for the synthesis of D-amino acid-containing peptides has been purified from the culture broth of Bacillus mycoides. The enzyme consisted of 4 subunits of 39 kDa, had an isoelectric point of 7.0, and showed its maximum activity at around 47 degrees C and pH 7.6. The enzyme activity was strongly depressed by phenylmethanesulfonyl fluoride, but not by penicillin G or ampicillin, suggesting that the protein is a serine enzyme lacking penicillin-binding ability. The enzyme hydrolyzed a variety of D- and L-amino acid methyl esters with concomitant formation of homooligomers from D-Phe, D-Trp, D-Tyr, and D-Asp(OCH(3)) methyl esters, but it did not act on the D- or L-amino acid amides tested. Incubation of a mixture of Ac-D-Phe-OMe and D-/L-Leu-NH(2) with the enzyme yielded Ac-D-Phe-D-/L-Leu-NH(2) together with Ac-D Phe-OH, the hydrolysate of the carboxyl component. To its credit, the enzyme failed to hydrolyze casein as well as peptides including diastereomers of diphenylalanine and dialanine, indicating that the enzyme would not cause secondary hydrolysis of once-formed peptides. These observations indicate the potential utility of the newly isolated enzyme for the synthesis of D-amino acid containing peptides. PMID- 11432788 TI - Effects of troponin T mutations in familial hypertrophic cardiomyopathy on regulatory functions of other troponin subunits. AB - We have previously shown that mutations in troponin T (TnT), which is associated with familial hypertrophic cardiomyopathy (HCM), cause an increase in the Ca(2+) sensitivity and a potentiation of cardiac muscle contraction. To gain further insight into the patho-physiological role of these mutations, four mutations (Arg92Gln, Phe110Ile, Glu244Asp, Arg278Cys) were introduced into recombinant human cardiac TnT, and the mutants were exchanged into isolated porcine cardiac myofibrils. The effects of mutations were tested on maximal ATPase activity, the inhibitory function of troponin I (TnI) in the absence of troponin C (TnC), and the neutralizing function of TnC. Arg92Gln, Phe110Ile, and Glu244Asp markedly impaired the inhibitory function of TnI. Arg278Cys also impaired the inhibitory function of TnI, but the effect was much smaller. Phe110Ile and Glu244Asp markedly enhanced the neutralizing function of TnC and potentiated the maximum ATPase activity. Arg92Gln and Arg278Cys only slightly enhanced the neutralizing function of TnC, and they conferred no potentiation on the maximum ATPase activity. These results indicate that mutations in TnT impair multiple processes of Ca(2+) regulation by troponin, and there are marked differences in the degree of impairment from mutation to mutation. PMID- 11432789 TI - A growing family of dual specificity phosphatases with low molecular masses. AB - Five putative dual specificity protein phosphatases (DSPs), designated LMW-DSP1, DSP4, -DSP6, -DSP10, and -DSP11, were cloned with a combination of RT-PCR and cDNA library screening strategies. Sequencing analysis revealed that all lacked the cdc25 homology domain that is conserved in most known DSPs/MAP kinase phosphatases (MKPs). LMW-DSP1 exhibited the highest similarity to plant DSPs. LMW DSP4 exhibited the highest similarity to human YVH1 and rat GKAP, but its C terminal region was much shorter than that of the human and rat clones. LMW-DSP6 was found to be identical to recently cloned TMDP, and LMW-DSP11 seemed to be a mouse ortholog of human VHR. LMW-DSP10 was found to have a DSP catalytic-like domain, but the critical cysteine residue for catalytic activity was missing. Recombinant LMW-DSP1, -DSP6, and -DSP11 exhibited obvious and strong activity against an artificial low molecular substrate, para-nitrophenyl phosphate (pNPP). Recombinant LMW-DSP4 exhibited slight but significant activity, whereas no activity was detected for LMW-DSP10. The phosphatase activity of the recombinant LMW-DSPs was inhibited by orthovanadate but not sodium fluoride. However, none of the DSPs could dephosphorylate MAP kinases such as ERK1, p38, and SAPK/JNK in transiently transfected COS7 cells under the conditions used. Northern blot analysis revealed that LMW-DSP1, -DSP6, -DSP10, and -DSP11 were specifically expressed in testis, while LMW-DSP4 was broadly expressed. The testis-specific expression and apparent absence of dephosphorylation action on MAP kinases suggest that LMW-DSP1, -DSP6, -DSP10, and -DSP11 play specific roles in testis. Taken together, it is conceivable that a distinct class of low molecular mass DSPs is present and plays a role in dephosphorylating unknown molecules other than MAP kinases. PMID- 11432790 TI - Role of 67 kDa cell surface laminin binding protein of Leishmania donovani in pathogenesis. AB - The role that interaction with laminin may play in Leishmania donovani infection was investigated. Binding of (125)I-radiolabeled laminin, in a liquid-phase assay, by the parasite was rapid, saturable, specific, reversible, and of high affinity. Using a Western blotting procedure, a 67 kDa laminin-binding protein (LBP) was identified from the membrane of both the promastigote and amastigote forms of L. donovani. Subsequently, the protein was purified by affinity chromatography. Immunofluorescence with a polyclonal antibody against LBP as well as flow cytometric analysis demonstrated its presence at the parasite surface. After stimulation with phorbol-12-myristate-13-acetate (PMA), U937 cells exhibited the ability to adhere to laminin and LBP specifically inhibited this adhesion. The reduced parasite adhesion after tunicamycin treatment suggested the importance of sugar residues in cell adhesion. Although co-administration of either laminin or LBP or anti LBP antibody reduced parasite virulence, resulting in a lower level of infection in the BALB/c mouse model, an in vitro macrophage culture-enhanced level of infection was observed in the case of laminin-coated parasites. The results collectively suggest a role for LBP in the interaction of the parasite with extracellular matrix elements, which may constitute a basis for the homing of the parasite to its physiological address. PMID- 11432791 TI - The second cytoplasmic loop of metabotropic glutamate receptor functions at the third loop position of rhodopsin. AB - G protein-coupled receptors identified so far are classified into at least three major families based on their amino acid sequences. For the family of receptors homologous to rhodopsin (family 1), the G protein activation mechanism has been investigated in detail, but much less for the receptors of other families. To functionally compare the G protein activation mechanism between rhodopsin and metabotropic glutamate receptor (mGluR), which belong to distinct families, we prepared a set of bovine rhodopsin mutants whose second or third cytoplasmic loop was replaced with either the second or third loop of Gi/Go- or Gq-coupled mGluR (mGluR6 or mGluR1). Among these mutants, the mutants in which the second or third loop was replaced with the corresponding loop of mGluR exhibited no G protein activation ability. In contrast, the mutant whose third loop was replaced with the second loop of Gi/Go-coupled mGluR6 efficiently activated Gi but not Gt: this activation profile is almost identical with those of the mutant rhodopsins whose third loop was replaced with those of the Gi/Go-coupled receptors in family 1 [Yamashita et al. (2000) J. Biol. Chem. 275, 34272-34279]. The mutant whose third loop was replaced with the second loop of Gq-coupled mGluR1 partially retained the Gi coupling ability of rhodopsin, which is in contrast to the fact that all the rhodopsin mutants having the third loops of Gq-coupled receptors in family 1 exhibit no detectable Gi activation. These results strongly suggest that the molecular architectures of rhodopsin and mGluR are different, although the G protein activation mechanism involving the cytoplasmic loops is common. PMID- 11432792 TI - Analysis of tyrosine phosphorylation-dependent protein-protein interactions in TrkB-mediated intracellular signaling using modified yeast two-hybrid system. AB - Activated receptor tyrosine kinases induce a large number of tyrosine phosphorylation-dependent protein-protein interactions through which they mediate their various ligand-exerted functions including regulation of proliferation, differentiation and survival. TrkB receptor tyrosine kinase activated by binding of brain-derived neurotrophic factor (BDNF) also stimulates various protein interactions in a tyrosine phosphorylation-dependent manner in neuronal cells. To examine tyrosine phosphorylation-dependent interactions stimulated by active TrkB, we developed a modified yeast two-hybrid system, which we call the yeast two-and-a-half-hybrid system. In this system, yeast was engineered to express a tyrosine kinase domain of TrkB as an effector, in addition to two fusion proteins with GAL4 DNA-binding and GAL4 activation domains as bait and prey proteins, respectively. Using this system with Shp2 as the bait, we demonstrated that Shp2 interacts directly with BIT/SHPS-1 (also called SIRP) and Grb2 depending on tyrosine phosphorylation mediated by TrkB. Furthermore, we screened an adult human brain cDNA library with the yeast two-and-a-half-hybrid system in order to identify other Shp2-binding proteins in TrkB-stimulated tyrosine phosphorylation signaling. We found that fibroblast growth factor receptor substrate 2beta (FRS2beta), also called SNT2, interacts with Shp2 dependently on TrkB-mediated tyrosine phosphorylation of FRS2beta/SNT2. Therefore, we show that the two-and-a half-hybrid system is a powerful tool for studying tyrosine phosphorylation dependent protein-protein interactions in intracellular signaling pathways stimulated by TrkB receptor tyrosine kinase. PMID- 11432793 TI - Albumin in the mg/l-range activates NF-kappaB in renal proximal tubule-derived cell lines via tyrosine kinases and protein kinase C. AB - Albuminuria represents one of the most unfavourable diagnostic factors for the prognosis of nephropathies. We investigated albumin-induced NF-kappaB activation and the potential contribution of tyrosine kinase and protein kinase C. Therefore we exposed proximal tubule-derived human (IHKE-1), opossum (OK) and porcine (LLC PK1) cell lines to serum albumin at concentrations of 10 - 500 mg/l. DNA-binding activity of NF-kappaB increased concentration-dependently in the presence of albumin. In OK and LLC-PK1 cells, NF-kappaB activity increased during the first 45 min and reached a plateau thereafter. In IHKE-1, cells NF-kappaB activity reached a plateau after 90 min with a maximum at 180 min exposure to albumin. The albumin-induced increase in NF-kappaB DNA-binding activity was inhibited by herbimycin A (tyrosine kinase inhibitor) and BIM (protein kinase C inhibitor). Reporter gene assays demonstrated that albumin stimulates NF-kappaB mediated reporter gene activation in LLC-PK1 cells, which was partially inhibited by herbimycin A and BIM. Our data indicate, that albumin exposure induces a rapid increase in NF-kappaB protein activity in renal proximal tubule cells of different species via a tyrosine kinase- and protein kinase C-dependent pathway, at concentrations occurring during mild glomerular injury. PMID- 11432794 TI - Reproductive assistance to HIV-discordant couples--the German approach. AB - An estimated 35 000 Germans are infected with HIV, and 75% of these are of childbearing age. Couples with one infected partner are faced with the risk of infection of the healthy partner. By using assisted reproduction techniques (ART) and virus-free sperm it is possible to fulfil the desire for children while minimising the risk of acquisition of infection. Therefore since 1991 in Germany ART is offered to couples of HIV-positive males and seronegative females. However, established programmes in different centres were temporarily suspended due to serious doubts about the reliability and safety of the sperm preparation technique (density gradient centrifugation and swim-up). -. Since 1997 processed sperm samples are routinely tested for HIV by highly sensitive detection methods and cryopreserved. Only if the test is negative are the processed spermatozoa used for homologous insemination, in-vitro fertilization or intracytoplasmatic sperm injection. This procedure removes most medical as well as legal considerations. At present 3 reproductive units in Germany offer reproductive assistance to HIV-discordant couples, and a further 9 institutions plan to establish that procedure. From 1991 to 1999, 54 women were treated by ART. 30 pregnancies occurred and 24 children were born. So far no maternal or infant infection was observed after treatment. PMID- 11432795 TI - Assessing the microcirculation in a burn wound by use of OPS imaging. AB - We report on the use of OPS imaging to visualize the microcirculation in a burn wound. This novel technique produces high quality images of the microcirculation and quantitative analysis of functional capillary density (FCD) was possible. FCD measured at day 3 following the injury was 11.2 +/- 4.6 (n/mm superset2; mean +/- SEM). During the initial phase of healing microcirculatory changes were characterized by a moderate but steady increase of FCD, which showed marked increase beginning from day 12 following the burn (16.6 +/- 6.9). Maximal FCD measured at day 23 (48.2 +/- 19.7) decreased from this point in time to finally reach 25.2 +/- 10.3 n/mm superset2 at the end of observation. OPS imaging allows for direct in vivo visualization and quantification of the microcirculation in burned skin. Our results of the use of OPS imaging in assessing the microcirculation in a burn wound appear promising, and we hope that this novel technique will allow to improve the knowledge of the dynamics of the microcirculation in the pathophysiology of thermal injury. PMID- 11432796 TI - The course of the occipital artery--an anatomical investigation for biopsy in suspected vasculitis. AB - BACKGROUND: The occipital artery can show an inflammation in cranial arteritis. For biopsy it is essential to know the course of the occipital artery. METHODS: In 6 randomly selected specimens of the head, the occipital artery and vein were sought and examined. In addition, the topographical proximity of the greater occipital nerve was considered. RESULTS: COURSE: The occipital artery followed a tortuous course, including an occasional hairpin bend in four out of 6 specimens. Lateral distance of the occipital artery from the external occipital protuberance of the occiput: The occipital artery runs at a mean distance of 3.92 cm on the right side and 4.4 cm on the left side from the midline. Variations of the course: A comparison between the right and left side showed a marked side difference in the course of the vessel. The extent of tortuosity varied distinctly. In most of the arteries, the angle between superior nuchal line and occipital artery was 90 degrees. External diameter: The average external diameter of the occipital artery (in the area where it crosses with the superior nuchal line) was 2.3 mm on the right side and 2.7 mm on the left side. CONCLUSION: Because the greater occipital nerve enters the subcutis below the external protuberance of the occiput and shortly afterwards crosses the occipital artery, we recommend to carry out the biopsy of the occipital artery between 1 to 3 cm above (cranially) and 4 to 5 cm lateral to the external protuberance. PMID- 11432797 TI - Efficacy and safety of an initial daily dosing regimen of interferon-alpha-2a in treatment-naive patients with chronic hepatitis C and HCV genotype 1. AB - Between January 1996 and February 1999 we treated two groups of patients with chronic hepatitis C and HCV genotype 1 according to different treatment regimens. 29 patients in group were treated in a prospective, open trial and received an initial dose of 6 MU Interferon-alpha-2a daily. In case of virologic response within the first 12 weeks of treatment a dosage reduction to 6 MU three times weekly was performed until the end of week 12 and 3 MU three times weekly thereafter until the completion of month 12. 35 patients in group 2 received 5 MU (Interferon-alpha-2b) or 6 MU (Interferon-alpha-2a) three times a week. If serum HCV RNA was negative after three months of treatment patients received 3 MU thrice weekly until completion of month 12. This regimen was considered standard therapy at the time of treatment. In both groups therapy was stopped in patients in whom HCV RNA remained detectable after 12 weeks of treatment or in patients who underwent virological breakthrough. The end point was a sustained virologic response defined as the absence of serum HCV RNA 6 months after treatment was completed. Primary response rates as defined by negative serum HCV RNA within the first 12 weeks of treatment were 59% in group 1 and 17% in group 2. Sustained response rates were 10% in group 1 and 3% in group 2. Initial daily dosing as in group 1 was therefore not associated with a higher sustained response rate compared to standard therapy. The safety profile was in accordance with the known side effects of interferon-alpha and was comparable in both treatment groups. PMID- 11432798 TI - Early antibiotic treatment and later asthma. AB - The reasons for the asthma epidemic are poorly understood. As the asthma prevalence follows the geographical and temporal trend of antibiotic use into clinical medicine, we examined a possible association in a population-based study of 2,512 children age 5-14 in East Germany. Wheezing was associated with increasing number of antibiotic courses (never versus one time odds ratio 1.9, P = 0.012, 2 to 5 times odds ratio 3.0, P<0.001 and more than 5 times, odds ratio 6.9, P<0.001) which was also seen for asthma diagnosis. The risk increased with earlier administration (never versus second year odds ratio 4.6, month 7-12 odds ratio 5.4 and birth until month 6 odds ratio 7.9, all P<0.001). Also non pulmonary treatment indication was associated with later wheezing (odds ratio 3.9, P<0.001). The most likely possible explanation is reverse causation indicating that frequent upper respiratory infections, an early symptom of asthma, are treated with antibiotics. Antibiotic therapy could also be a proxy of another closely associated genetic or environmental factor. The high dose effect, the time dependency of the administration and the effect by non-pulmonary indications raises the possibility that early antibiotic treatment could itself be related to later asthma. PMID- 11432799 TI - Therapy of chronic hepatitis B and C and treatment options in HCV-HIV co infection--European Conference on Infectious Diseases and Tropical Medicine, EuCID 2001, 3-6 May 2001, Leipzig. AB - Viral hepatitis features with an incidence of 500 Mio infected individuals worldwide. Chronification of the infection as seen in hepatitis C, B, and D may lead to liver cirrhosis and its associated complications in later stages of the disease. Especially the chronification of acute hepatitis C is observed in a majority of cases (50-80%). Chronic Hepatitis C (cHC) should be treated in generally when elevated serum concentrations of liver enzymes are found or symptoms of disease occur. Treatment goals are viral elimination, improve in histology, prevention of HCC, and a better quality of life for the patients. As HCV and HIV share the same transmission routes, a relatively high rate of HCV-HIV co-infection is observed. Co-infection is characterized by a more progressive natural course of HCV-infection, leading to an increased mortality due to liver failure in the afflicted patients. The development of treatment options for anti HCV-specific therapy in dually infected patients is urgently needed. The European Conference on Infectious Diseases and Tropical Medicine, EuCID 2001, which took place in May 3-6, 2001, in Leipzig covered aspects of viral hepatitis and its treatment in several sessions and gave latest results on treatment with interferon-alfa, lamivudine, and adefovir in hepatitis B as well as interferon alfa, pegylated interferon-alfa, and ribavirin in hepatitis C and HCV-HIV co infection respectively. PMID- 11432800 TI - Bisoprolol treatment for cyanotic spells--a 69-year-old female with uncorrected pentalogy of Fallot. AB - Pentalogy of Fallot is a rare cyanotic congenital heart disease characterized by biventricular origin of the aorta above a large ventricular septal defect, obstruction of the pulmonary outflow, right ventricular hypertrophy (tetralogy of Fallot), and an atrial septal defect. Mortality due to syncope, arrhythmia or congestive heart failure is about 75% by the age of 10 years and 97% by the age of 40. We report a 69-year old female with uncorrected pentalogy of Fallot who suffered from recurrent syncope during classic cyanotic spells. Treatment with bisoprolol is recommended in symptomatic children and also improved symptoms in our patient. We conclude that single patients with pentalogy of Fallot reach the seventh decade and that cyanotic congenital heart disease is a rare cause of syncope in the elderly. PMID- 11432801 TI - Mechanism of block of single protopores of the Torpedo chloride channel ClC-0 by 2-(p-chlorophenoxy)butyric acid (CPB). AB - We investigated in detail the mechanism of inhibition by the S(-) enantiomer of 2 (p-chlorophenoxy)butyric acid (CPB) of the Torpedo Cl(-)channel, ClC-0. The substance has been previously shown to inhibit the homologous skeletal muscle channel, CLC-1. ClC-0 is a homodimer with probably two independently gated protopores that are conductive only if an additional common gate is open. As a simplification, we used a mutant of ClC-0 (C212S) that has the common gate "locked open" (Lin, Y.W., C.W. Lin, and T.Y. Chen. 1999. J. Gen. Physiol. 114:1 12). CPB inhibits C212S currents only when applied to the cytoplasmic side, and single-channel recordings at voltages (V) between -120 and -80 mV demonstrate that it acts independently on individual protopores by introducing a long-lived nonconductive state with no effect on the conductance and little effect on the lifetime of the open state. Steady-state macroscopic currents at -140 mV are half inhibited by approximately 0.5 mM CPB, but the inhibition decreases with V and vanishes for V > or = 40 mV. Relaxations of CPB inhibition after voltage steps are seen in the current responses as an additional exponential component that is much slower than the gating of drug-free protopores. For V = 60 mV) with an IC50 of approximately 30-40 mM. Altogether, these findings support a model for the mechanism of CPB inhibition in which the drug competes with Cl(-) for binding to a site of the pore where it blocks permeation. CPB binds preferentially to closed channels, and thereby also strongly alters the gating of the single protopore. Since the affinity of CPB for open WT pores is extremely low, we cannot decide in this case if it acts also as an open pore blocker. However, the experiments with the mutant K519E strongly support this interpretation. CPB block may become a useful tool to study the pore of ClC channels. As a first application, our results provide additional evidence for a double-barreled structure of ClC-0 and ClC-1. PMID- 11432802 TI - Cdk5 on the brain. AB - Mammalian brains are highly compartmentalized into groups of functionally specialized neurons. Cell migration and neurite outgrowth must be tightly orchestrated to achieve this level of organization. A small serine/threonine kinase that shows homology to cyclin-dependent kinases (Cdks) has emerged as an important regulator of neuronal migration. Cdk5, unlike other Cdks, is not regulated by cyclins, and its activity is primarily detected in postmitotic neurons in developing and adult nervous systems. This review describes work indicating that Cdk5 links extracellular signaling pathways and cytoskeletal/membrane systems to direct neuronal migration, axon growth, and possibly neurosecretion. Despite its importance, unchecked Cdk5 activity is toxic to neurons, and may underlie some of the pathologies associated with neurodegenerative disorders such as Alzheimer's disease and amyotrophic lateral sclerosis. PMID- 11432803 TI - Cyclin-dependent kinase (cdk) inhibitors/cdk4/cdk2 complexes in early stages of mouse mammary preneoplasia. AB - The level of circulating ovarian hormones (estrogen and progesterone) alone or in combination with pituitary hormones have a potent mitogenic impact in the normal mammary gland, and they also play a pivotal role in the development and progression of mammary carcinoma. The differential effects of hormones on the molecular components of cyclin-dependent kinase (cdk) complexes in mammary epithelium of the hormone-dependent ductal outgrowth line, EL11, and the hormone independent alveolar outgrowth line, TM2L, were the focus of this study. The two outgrowth lines, which represent early stages in mammary hyperplasia, were compared with normal mammary gland at different hormonal conditions: control, hormone stimulated by pituitary isograft, and hormone depleted by ovariectomy. Hormonal stimulation by a pituitary isograft resulted in DNA synthesis and lobuloalveolar development of normal mammary ducts, DNA synthesis but no lobuloalveolar development in the EL11 ductal outgrowth, and no changes either in DNA synthesis or in lobuloalveolar morphology in the TM2L outgrowth. The levels of cdk4- and cyclin D1-associated kinase activities were correlated with cell proliferation in only the alveolar phenotypes (i.e., in only hormonally stimulated normal virgin gland and in alveolar mammary outgrowth), whereas cyclin D2-dependent kinase activity was correlated with cell proliferation in only the alveolar preneoplasia. p16(INK4a) and p21(Cip1) protein levels were decreased at the earliest stages of preneoplasia, i.e., at immortalization, and were independent from changes in cyclin D1, which occurred later in preneoplasia. Although all cdk inhibitors changed in concordance with hormonal status reflected by proliferation levels, p27(Kip1) was the only cdk inhibitor that was up regulated at the earliest stages of preneoplasia and may have a unique role in blocking alveolar differentiation in response to the loss of one or more of the cell cycle-negative regulators. We hypothesize that up-regulation of p27(Kip1) prevents immortalized ductal outgrowths (EL11) from progressing to the neoplastic state, even under hormonal stimulation. PMID- 11432804 TI - The recruitment of Fas-associated death domain/caspase-8 in Ras-induced apoptosis. AB - Oncogenic Ras induces cells to undergo apoptosis after inhibition of protein kinase C (PKC) activity. The integration of differential signaling pathways is required for full execution of apoptosis. In this study, we used Jurkat as well as Fas/FADD-defective cell lines expressing v-ras to determine the upstream elements required for activation of the caspase cascade in PKC/Ras-mediated apoptosis. During this Ras-induced apoptotic process, caspase-8 was activated, possibly through its binding to Fas-associated death domain (FADD), in Jurkat/ras and Jurkat/Fas(m)/ras cells but not in Jurkat/FADD(m)/ras cells. c-Jun NH(2) terminal kinase (JNK) was activated in all three cell lines expressing ras in response to apoptotic stimulation. Suppression of JNK by dn-JNK1 blocked the interaction of FADD and caspase-8 and partially protected Jurkat/ras and Jurkat/Fas(m)/ras cells from apoptosis. However, dn-JNK1 had no effect on PKC/Ras induced apoptosis in Jurkat/FADD(m)/ras cells. The results indicate that FADD/caspase-8 signaling is involved in PKC/Ras-mediated apoptosis, and JNK may be an upstream effector of caspase activation. PMID- 11432805 TI - Identification of epidermal growth factor receptor- Grb2-associated binder-1-SHP 2 complex formation and its functional loss during neoplastic cell progression. AB - The adaptor protein Grb2-associated binder-1 (Gab1) is known to bind to the SHP-2 tyrosine phosphatase on epidermal growth factor (EGF) receptor stimulation. To clarify the roles of these two proteins in EGF receptor (EGFR) signaling and determine their possible alteration during neoplastic cell progression, we studied these proteins in a Syrian hamster embryo (SHE) cell line model of neoplastic progression. Specifically, we used asbestos-transformed SHE fibroblasts: the 10W+8 clone, which is immortal but nontumorigenic; and the 10W2T clone, which is tumorigenic. Gab1 was detected, and the EGF-dependent formation of the EGFR-Gab1-SHP-2 complex was observed in 10W+8 cells. After cloning hamster Gab1 cDNA, exogenous expression of Gab1 significantly enhanced EGF-dependent mitogenic activity in 10W+8 cells. On the other hand, Gab1 was not detected in 10W2T cells, and the EGF-dependent association of SHP-2 with EGFR was also absent. Exogenous Gab1 expression in transfected 10W2T cells restored the EGF dependent association of SHP-2 with EGFR, although it only showed a marginal effect on EGF-dependent mitogenic activity. Thus, Gab1 plays a pivotal role in the EGFR signaling pathway via the formation of the EGFR-Gab1-SHP-2 complex, and alteration in the expression and function of Gab1 is implicated in the neoplastic progression of SHE cells. PMID- 11432806 TI - Transcriptional regulation of the vitamin D(3) receptor gene by ZEB. AB - The hormone 1,25-dihydroxyvitamin D(3) influences the growth and differentiation of a number of cell types. The functions of 1,25-dihydroxyvitamin D(3) are mediated through the vitamin D(3) receptor (VDR); therefore, an understanding of the regulation of VDR expression is important when considering the molecular mechanisms of differentiation induced by vitamin D(3) and its analogues. ZEB, a Kruppel-type transcription factor known to repress the transcription of several genes, binds to two sites within the VDR promoter and activates the transcription of this receptor in a cell-specific manner. Transfection of ZEB into SW620 colon carcinoma cells results in an up-regulation of the expression of endogenous VDR, confirming the role of ZEB in the transcriptional activation of the VDR gene. The expression of VDR is also induced by c-MYB; thus, ZEB and c-MYB may modulate the levels of VDR expression during differentiation in embryonal development, as well as in cancer cells. PMID- 11432807 TI - Varying functions of specific major histocompatibility class II transactivator promoter III and IV elements in melanoma cell lines. AB - Melanoma cells commonly express MHC class II molecules constitutively. This is a rare, or possibly unique, phenotype for a nonprofessional antigen-presenting cell, where MHC class II expression ordinarily occurs only after IFN-gamma treatment. Despite the fact that constitutive expression of MHC class II on melanoma cells has been observed for decades and that the regulation of the MHC class II genes is well understood for many different cell types, there is no data regarding the basis for constitutive MHC class II expression in melanoma cells. Here we report that MHC class II expression in melanoma cells can be traced to constitutive expression of the class II transactivator protein (CIITA), which mediates both IFN-gamma-inducible and -constitutive MHC class II expression in all other cell types. In addition, we determined that constitutive CIITA expression is the result of the activation of both the B cell-specific CIITA promoter III and the IFN-gamma-inducible CIITA promoter IV, the latter of which previously has never been known to function as a constitutive promoter in any cell type. The recently described B cell-related ARE-1 activity is important for promoter III activation in the melanoma cells. Constitutive promoter IV activation involves the IFN regulatory factor element (IRF-E), which binds members of the IRF family of proteins, although the major, IFN-gamma inducible member of this family, IRF-1, is not constitutively expressed in these cells. In cells with constitutively active promoter IV, the promoter IV IRF-E is most likely activated by IRF-2. The relevance of these results to the pathway of melanoma development is discussed. PMID- 11432809 TI - Sex-associated hormones and immunity to protozoan parasites. AB - Numerous epidemiological and clinical studies have noted differences in the incidence and severity of parasitic diseases between males and females. Although in some instances this may be due to gender-associated differences in behavior, there is overwhelming evidence that sex-associated hormones can also modulate immune responses and consequently directly influence the outcome of parasitic infection. Animal models of disease can often recreate the gender-dependent differences observed in humans, and the role of sex-associated hormones can be confirmed by experimentally altering their levels. Under normal circumstances, levels of sex hormones not only differ between males and females but vary according to age. Furthermore, not only are females of reproductive age subject to the regular hormonal cycles which control ovulation, they are also exposed to dramatically altered levels during pregnancy. It is thus not surprising that the severity of many diseases, including those caused by parasites, has been shown to be affected by one or more of these circumstances. In addition, infection with many pathogens has been shown to have an adverse influence on pregnancy. In this article we review the impact of sex-associated hormones on the immune system and the development and maintenance of immunity to the intracellular protozoan parasites Toxoplasma gondii, Plasmodium spp., and Leishmania spp. PMID- 11432808 TI - Biology of Giardia lamblia. AB - Giardia lamblia is a common cause of diarrhea in humans and other mammals throughout the world. It can be distinguished from other Giardia species by light or electron microscopy. The two major genotypes of G. lamblia that infect humans are so different genetically and biologically that they may warrant separate species or subspecies designations. Trophozoites have nuclei and a well-developed cytoskeleton but lack mitochondria, peroxisomes, and the components of oxidative phosphorylation. They have an endomembrane system with at least some characteristics of the Golgi complex and encoplasmic reticulum, which becomes more extensive in encysting organisms. The primitive nature of the organelles and metabolism, as well as small-subunit rRNA phylogeny, has led to the proposal that Giardia spp. are among the most primitive eukaryotes. G. lamblia probably has a ploidy of 4 and a genome size of approximately 10 to 12 Mb divided among five chromosomes. Most genes have short 5' and 3' untranslated regions and promoter regions that are near the initiation codon. Trophozoites exhibit antigenic variation of an extensive repertoire of cysteine-rich variant-specific surface proteins. Expression is allele specific, and changes in expression from one vsp gene to another have not been associated with sequence alterations or gene rearrangements. The Giardia genome project promises to greatly increase our understanding of this interesting and enigmatic organism. PMID- 11432811 TI - Borna disease virus and human disease. AB - The biology of Borna disease virus (BDV) strongly supports the likelihood of human infection with BDV or a variant of BDV. Thus far, the evidence supporting BDV infection in humans has initiated much controversy among basic and clinical scientists; only time and additional research will support or refute the hypothesis of human BDV infection. Until an assay of acceptable specificity and sensitivity has been developed, validated, and used to document human BDV infection, scientists cannot reasonably begin to associate BDV infection with specific disease syndromes. Clinical studies seeking causal associations between BDV infection and specific diseases must ensure the proper identification of the BDV infection status of patients and control subjects by using a validated, highly sensitive, and highly specific assay (or series of assays). For clinical studies, a highly sensitive "screening" test followed by a highly specific confirmatory test will be of significant benefit. Although it is possible to formulate hypotheses about the clinical outcomes of human BDV infection based on animal model work, to date no human disease has been causally linked to human BDV infection. Scientists all over the world are actively pursuing these issues, and with continuing advances in clinical and basic BDV research, the answers cannot be far away. PMID- 11432810 TI - Mycobacterium avium subsp. paratuberculosis in Veterinary Medicine. AB - Mycobacterium avium subsp. paratuberculosis (basonym M. paratuberculosis) is the etiologic agent of a severe gastroenteritis in ruminants known as Johne's disease. Economic losses to the cattle industry in the United States are staggering, reaching $1.5 billion annually. A potential pathogenic role in humans in the etiology of Crohn's disease is under investigation. In this article, we review the epidemiology, pathogenesis, diagnostics, and disease control measures of this important veterinary pathogen. We emphasize molecular genetic aspects including the description of markers used for strain identification, diagnostics, and phylogenetic analysis. Recent important advances in the development of animal models and genetic systems to study M. paratuberculosis virulence determinants are also discussed. We conclude with proposals for the applications of these models and recombinant technology to the development of diagnostic, control, and therapeutic measures. PMID- 11432812 TI - Nosocomial spread of viral disease. AB - Viruses are important causes of nosocomial infection, but the fact that hospital outbreaks often result from introduction(s) from community-based epidemics, together with the need to initiate specific laboratory testing, means that there are usually insufficient data to allow the monitoring of trends in incidences. The most important defenses against nosocomial transmission of viruses are detailed and continuing education of staff and strict adherence to infection control policies. Protocols must be available to assist in the management of patients with suspected or confirmed viral infection in the health care setting. In this review, we present details on general measures to prevent the spread of viral infection in hospitals and other health care environments. These include principles of accommodation of infected patients and approaches to good hygiene and patient management. They provide detail on individual viral diseases accompanied in each case with specific information on control of the infection and, where appropriate, details of preventive and therapeutic measures. The important areas of nosocomial infection due to blood-borne viruses have been extensively reviewed previously and are summarized here briefly, with citation of selected review articles. Human prion diseases, which present management problems very different from those of viral infection, are not included. PMID- 11432813 TI - Role of genomic typing in taxonomy, evolutionary genetics, and microbial epidemiology. AB - Currently, genetic typing of microorganisms is widely used in several major fields of microbiological research. Taxonomy, research aimed at elucidation of evolutionary dynamics or phylogenetic relationships, population genetics of microorganisms, and microbial epidemiology all rely on genetic typing data for discrimination between genotypes. Apart from being an essential component of these fundamental sciences, microbial typing clearly affects several areas of applied microbiological research. The epidemiological investigation of outbreaks of infectious diseases and the measurement of genetic diversity in relation to relevant biological properties such as pathogenicity, drug resistance, and biodegradation capacities are obvious examples. The diversity among nucleic acid molecules provides the basic information for all fields described above. However, researchers in various disciplines tend to use different vocabularies, a wide variety of different experimental methods to monitor genetic variation, and sometimes widely differing modes of data processing and interpretation. The aim of the present review is to summarize the technological and fundamental concepts used in microbial taxonomy, evolutionary genetics, and epidemiology. Information on the nomenclature used in the different fields of research is provided, descriptions of the diverse genetic typing procedures are presented, and examples of both conceptual and technological research developments for Escherichia coli are included. Recommendations for unification of the different fields through standardization of laboratory techniques are made. PMID- 11432814 TI - Whipple's disease and "Tropheryma whippelii". AB - Whipple's disease is a rare bacterial infection that may involve any organ system in the body. It occurs primarily in Caucasian males older than 40 years. The gastrointestinal tract is the most frequently involved organ, with manifestations such as abdominal pain, malabsorption syndrome with diarrhea, and weight loss. Other signs include low-grade fever, lymphadenopathy, skin hyperpigmentation, endocarditis, pleuritis, seronegative arthritis, uveitis, spondylodiscitis, and neurological manifestations, and these signs may occur in the absence of gastrointestinal manifestations. Due to the wide variability of manifestations, clinical diagnosis is very difficult and is often made only years or even decades after the initial symptoms have appeared. Trimethoprim-sulfamethoxazole for at least 1 year is usually considered adequate to eradicate the infection. The microbiological diagnosis of this insidious disease is rendered difficult by the virtual lack of culture and serodiagnostic methods. It is usually based on the demonstration of periodic acid-Schiff-positive particles in infected tissues and/or the presence of bacteria with an unusual trilaminar cell wall ultrastructure by electron microscopy. Recently, the Whipple bacteria have been characterized at the molecular level by amplification of their 16S rRNA gene(s). Phylogenetic analysis of these sequences revealed a new bacterial species related to the actinomycete branch which was named "Tropheryma whippelli." Based on its unique 16S ribosomal DNA (rDNA) sequence, species-specific primers were selected for the detection of the organism in clinical specimens by PCR. This technique is currently used as one of the standard methods for establishing the diagnosis of Whipple's disease. Specific and broad-spectrum PCR amplifications mainly but not exclusively from extraintestinal specimens have significantly improved diagnosis, being more sensitive than histopathologic analysis. However, "T. whippelii" DNA has also been found in persons without clinical and histological evidence of Whipple's disease. It is unclear whether these patients are true asymptomatic carriers or whether differences in virulence exist among strains of "T. whippelii" that might account for the variable clinical manifestations. So far, six different "T. whippelii" subtypes have been found by analysis of their 16S 23S rDNA spacer region. Further studies of the pathogen "T. whippelii" as well as the host immune response are needed to fully understand this fascinating disease. The recent cultivation of the organisms is a promising major step in this direction. PMID- 11432816 TI - Control of chromosomal DNA replication in the early Xenopus embryo. PMID- 11432817 TI - Nuclear interpretation of Dpp signaling in Drosophila. AB - Signaling by Decapentaplegic (Dpp), a member of the TGFbeta superfamily of signaling molecules similar to vertebrate BMP2 and BMP4, has been implicated in many developmental processes in Drosophila melanogaster. Notably, Dpp acts as a long-range morphogen during imaginal disc growth and patterning. Genetic approaches led to the identification of a number of gene products that constitute the core signaling pathway. In addition to the ligand-activated heteromeric receptor complex and the signal-transducing intracellular Smad proteins, Dpp signaling requires two nuclear proteins, Schnurri (Shn) and Brinker (Brk), to prime cells for Dpp responsiveness. A complex interplay between the nuclear factors involved in Dpp signaling appears to control the transcriptional readout of the Dpp morphogen gradient. It remains to be seen whether similar molecular mechanisms operate in the nucleus in vertebrate systems. PMID- 11432818 TI - Crystal structure of a superantigen bound to MHC class II displays zinc and peptide dependence. AB - The three-dimensional structure of a bacterial superantigen, Staphylococcus aureus enterotoxin H (SEH), bound to human major histocompatibility complex (MHC) class II (HLA-DR1) has been determined by X-ray crystallography to 2.6 A resolution (1HXY). The superantigen binds on top of HLA-DR1 in a completely different way from earlier co-crystallized superantigens from S.aureus. SEH interacts with high affinity through a zinc ion with the beta1 chain of HLA-DR1 and also with the peptide presented by HLA-DR1. The structure suggests that all superantigens interacting with MHC class II in a zinc-dependent manner present the superantigen in a common way. This suggests a new model for ternary complex formation with the T-cell receptor (TCR), in which a contact between the TCR and the MHC class II is unlikely. PMID- 11432819 TI - Three-dimensional structure of the lithostathine protofibril, a protein involved in Alzheimer's disease. AB - Neurodegenerative diseases are characterized by the presence of filamentous aggregates of proteins. We previously established that lithostathine is a protein overexpressed in the pre-clinical stages of Alzheimer's disease. Furthermore, it is present in the pathognomonic lesions associated with Alzheimer's disease. After self-proteolysis, the N-terminally truncated form of lithostathine leads to the formation of fibrillar aggregates. Here we observed using atomic force microscopy that these aggregates consisted of a network of protofibrils, each of which had a twisted appearance. Electron microscopy and image analysis showed that this twisted protofibril has a quadruple helical structure. Three dimensional X-ray structural data and the results of biochemical experiments showed that when forming a protofibril, lithostathine was first assembled via lateral hydrophobic interactions into a tetramer. Each tetramer then linked up with another tetramer as the result of longitudinal electrostatic interactions. All these results were used to build a structural model for the lithostathine protofibril called the quadruple-helical filament (QHF-litho). In conclusion, lithostathine strongly resembles the prion protein in its dramatic proteolysis and amyloid proteins in its ability to form fibrils. PMID- 11432820 TI - Specificity of GlcNAc-PI de-N-acetylase of GPI biosynthesis and synthesis of parasite-specific suicide substrate inhibitors. AB - The substrate specificities of Trypanosoma brucei and human (HeLa) GlcNAc-PI de-N acetylases were determined using 24 substrate analogues. The results show the following. (i) The de-N-acetylases show little specificity for the lipid moiety of GlcNAc-PI. (ii) The 3'-OH group of the GlcNAc residue is essential for substrate recognition whereas the 6'-OH group is dispensable and the 4'-OH, while not required for recognition, cannot be epimerized or substituted. (iii) The parasite enzyme can act on analogues containing betaGlcNAc or aromatic N-acyl groups, whereas the human enzyme cannot. (iv) Three GlcNR-PI analogues are de-N acetylase inhibitors, one of which is a suicide inhibitor. (v) The suicide inhibitor most likely forms a carbamate or thiocarbamate ester to an active site hydroxy-amino acid or Cys or residue such that inhibition is reversed by certain nucleophiles. These and previous results were used to design two potent (IC50 = 8 nM) parasite-specific suicide substrate inhibitors. These are potential lead compounds for the development of anti-protozoan parasite drugs. PMID- 11432815 TI - Listeria pathogenesis and molecular virulence determinants. AB - The gram-positive bacterium Listeria monocytogenes is the causative agent of listeriosis, a highly fatal opportunistic foodborne infection. Pregnant women, neonates, the elderly, and debilitated or immunocompromised patients in general are predominantly affected, although the disease can also develop in normal individuals. Clinical manifestations of invasive listeriosis are usually severe and include abortion, sepsis, and meningoencephalitis. Listeriosis can also manifest as a febrile gastroenteritis syndrome. In addition to humans, L. monocytogenes affects many vertebrate species, including birds. Listeria ivanovii, a second pathogenic species of the genus, is specific for ruminants. Our current view of the pathophysiology of listeriosis derives largely from studies with the mouse infection model. Pathogenic listeriae enter the host primarily through the intestine. The liver is thought to be their first target organ after intestinal translocation. In the liver, listeriae actively multiply until the infection is controlled by a cell-mediated immune response. This initial, subclinical step of listeriosis is thought to be common due to the frequent presence of pathogenic L. monocytogenes in food. In normal individuals, the continual exposure to listerial antigens probably contributes to the maintenance of anti-Listeria memory T cells. However, in debilitated and immunocompromised patients, the unrestricted proliferation of listeriae in the liver may result in prolonged low-level bacteremia, leading to invasion of the preferred secondary target organs (the brain and the gravid uterus) and to overt clinical disease. L. monocytogenes and L. ivanovii are facultative intracellular parasites able to survive in macrophages and to invade a variety of normally nonphagocytic cells, such as epithelial cells, hepatocytes, and endothelial cells. In all these cell types, pathogenic listeriae go through an intracellular life cycle involving early escape from the phagocytic vacuole, rapid intracytoplasmic multiplication, bacterially induced actin-based motility, and direct spread to neighboring cells, in which they reinitiate the cycle. In this way, listeriae disseminate in host tissues sheltered from the humoral arm of the immune system. Over the last 15 years, a number of virulence factors involved in key steps of this intracellular life cycle have been identified. This review describes in detail the molecular determinants of Listeria virulence and their mechanism of action and summarizes the current knowledge on the pathophysiology of listeriosis and the cell biology and host cell responses to Listeria infection. This article provides an updated perspective of the development of our understanding of Listeria pathogenesis from the first molecular genetic analyses of virulence mechanisms reported in 1985 until the start of the genomic era of Listeria research. PMID- 11432821 TI - Requirement for C3G-dependent Rap1 activation for cell adhesion and embryogenesis. AB - C3G is a guanine nucleotide exchange factor (GEF) for Rap1, and is activated via Crk adaptor protein. To understand the physiological role of C3G, we generated C3G knockout mice. C3G(-/-) homozygous mice died before embryonic day 7.5. The lethality was rescued by the expression of the human C3G transgene, which could be excised upon the expression of Cre recombinase. From the embryo of this mouse, we prepared fibroblast cell lines, MEF-hC3G. Expression of Cre abolished the expression of C3G in MEF-hC3G and inhibited cell adhesion-induced activation of Rap1. The Cre-expressing MEF-hC3G showed impaired cell adhesion, delayed cell spreading and accelerated cell migration. The accelerated cell migration was suppressed by the expression of active Rap1, Rap2 and R-Ras. Expression of Epac and CalDAG-GEFI, GEFs for Rap1, also suppressed the accelerated migration of the C3G-deficient cells. This observation indicated that Rap1 activation was sufficient to complement the C3G deficiency. In conclusion, C3G-dependent activation of Rap1 is required for adhesion and spreading of embryonic fibroblasts and for the early embryogenesis of the mouse. PMID- 11432822 TI - N-arginine dibasic convertase is a specific receptor for heparin-binding EGF-like growth factor that mediates cell migration. AB - Heparin-binding epidermal growth factor-like growth factor (HB-EGF), a mitogen and chemotactic factor, binds to two receptor tyrosine kinases, erbB1 and erbB4. Now we demonstrate that HB-EGF also binds to a novel 140 kDa receptor on MDA-MB 453 cells. Purification of this receptor showed it to be identical to N-arginine dibasic convertase (NRDc), a metalloendopeptidase of the M16 family. Binding to cell surface NRDc and NRDc in solution was highly specific for HB-EGF among EGF family members. When overexpressed in cells, NRDc enhanced their migration in response to HB-EGF but not to EGF. Conversely, inhibition of endogenous NRDc expression in cells by antisense morpholino oligonucleotides inhibited HB-EGF induced cell migration. Anti-erbB1 neutralizing antibodies completely abrogated the ability of NRDc to enhance HB-EGF-dependent migration, demonstrating that this NRDc activity was dependent on erbB1 signaling. Although NRDc is a metalloproteinase, enzymatic activity was not required for HB-EGF binding or enhancement of cell migration; neither did NRDc cleave HB-EGF. Together, these results suggest that NRDc is a novel specific receptor for HB-EGF that modulates HB-EGF-induced cell migration via erbB1. PMID- 11432823 TI - Scrapie strains maintain biological phenotypes on propagation in a cell line in culture. AB - Bovine spongiform encephalopathy (BSE) and its human equivalent, variant Creutzfeldt-Jakob disease (vCJD), are caused by the same strain of infectious agent, which is similar to, but distinct from, >20 strains of their sheep scrapie homologue. A better understanding of the molecular strain determinants could be obtained from cells in monoculture than from whole animal studies where different cell targeting is commonly a strain-related feature. Although a few cell types can be infected with different strains, the phenotypes of the emergent strains have not been studied. We have cured the scrapie-infected, clonal SMB cell line with pentosan sulfate, stably re-infected it with a different strain of scrapie and shown that biological properties and prion protein profiles characteristic of each original strain are propagated faithfully in this single non-neuronal cell type. These findings attest to the fact that scrapie strain determinants are stable and host-independent in isolated cells. PMID- 11432824 TI - Lysine 188 substitutions convert the pattern of proteasome activation by REGgamma to that of REGs alpha and beta. AB - 11S REGs (PA28s) are multimeric rings that bind proteasomes and stimulate peptide hydrolysis. Whereas REGalpha activates proteasomal hydrolysis of peptides with hydrophobic, acidic or basic residues in the P1 position, REGgamma only activates cleavage after basic residues. We have isolated REGgamma mutants capable of activating the hydrolysis of fluorogenic peptides diagnostic for all three active proteasome beta subunits. The most robust REGgamma specificity mutants involve substitution of Glu or Asp for Lys188. REGgamma(K188E/D) variants are virtually identical to REGalpha in proteasome activation but assemble into less stable heptamers/hexamers. Based on the REGalpha crystal structure, Lys188 of REGgamma faces the aqueous channel through the heptamer, raising the possibility that REG channels function as substrate-selective gates. However, covalent modification of proteasome chymotrypsin-like subunits by 125I-YL3-VS demonstrates that REGgamma(K188E)'s activation of all three proteasome active sites is not due to relaxed gating. We propose that decreased stability of REGgamma(K188E) heptamers allows them to change conformation upon proteasome binding, thus relieving inhibition of the CT and PGPH sites normally imposed by the wild-type REGgamma molecule. PMID- 11432825 TI - Analysis of heterodimer formation by Xklp3A/B, a newly cloned kinesin-II from Xenopus laevis. AB - kinesin-II motor proteins are composed of two different kinesin-like motor proteins and one cargo binding subunit. Here we report the cloning of a new member of the kinesin-II superfamily, Xklp3A from Xenopus laevis, which forms a heterodimeric complex with Xklp3B. The heterodimer formation properties between Xklp3A and B have been tested in vitro using reticulocyte lysate expression and immunoprecipitation. To this end we produced a series of Xklp3A and B constructs of varying length and tested their propensity for heterodimer formation. We could demonstrate that, in contrast to conventional kinesin, the critical domains for heterodimer formation in Xklp3A/B are located at the C-terminal end of the stalk. Neither the neck nor the highly charged stretches after the neck region, which are typical of kinesins-II, are required for heterodimer formation, nor do they prevent homodimer formation. Dimerization is controlled by a cooperative mechanism between the C-terminal coiled-coil segments. Classical trigger sites were not identified. The critical regions for dimerization exhibit a very high degree of sequence conservation among equivalent members of the kinesin-II family. PMID- 11432826 TI - Vps45p stabilizes the syntaxin homologue Tlg2p and positively regulates SNARE complex formation. AB - Sec1p-like/Munc-18 (SM) proteins bind to t-SNAREs and inhibit ternary complex formation. Paradoxically, the absence of SM proteins does not result in constitutive membrane fusion. Here, we show that in yeast cells lacking the SM protein Vps45p, the t-SNARE Tlg2p is down-regulated, to undetectable levels, by rapid proteasomal degradation. In the absence of Vps45p, Tlg2p can be stabilized through abolition of proteasome activity. Surprisingly, the stabilized Tlg2p was targeted to the correct intracellular location. However, the stabilized Tlg2p is non-functional and unable to bind its cognate SNARE binding partners, Tlg1p and Vti1p, in the absence of Vps45p. A truncation mutant lacking the first 230 residues of Tlg2p no longer bound Vps45p but was able to form complexes with Tlg1p and Vti1p in the absence of the SM protein. These data provide us with two valuable insights into the function of SM proteins. First, SM proteins act as chaperone-like molecules for their cognate t-SNAREs. Secondly, SM proteins play an essential role in the activation process allowing their cognate t-SNARE to participate in ternary complex formation. PMID- 11432827 TI - Fission yeast ch-TOG/XMAP215 homologue Alp14 connects mitotic spindles with the kinetochore and is a component of the Mad2-dependent spindle checkpoint. AB - The TOG/XMAP215-related proteins play a role in microtubule dynamics at its plus end. Fission yeast Alp14, a newly identified TOG/XMAP215 family protein, is essential for proper chromosome segregation in concert with a second homologue Dis1. We show that the alp14 mutant fails to progress towards normal bipolar spindle formation. Intriguingly, Alp14 itself is a component of the Mad2 dependent spindle checkpoint cascade, as upon addition of microtubule destabilizing drugs the alp14 mutant is incapable of maintaining high H1 kinase activity, which results in securin destruction and premature chromosome separation. Live imaging of Alp14-green fluorescent protein shows that during mitosis, Alp14 is associated with the peripheral region of the kinetochores as well as with the spindle poles. This is supported by ChIP (chromatin immunoprecipitation) and overlapping localization with the kinetochore marker Mis6. An intact spindle is required for Alp14 localization to the kinetochore periphery, but not to the poles. These results indicate that the TOG/XMAP215 family may play a central role as a bridge between the kinetochores and the plus end of pole to chromosome microtubules. PMID- 11432828 TI - Cell-autonomous and non-cell-autonomous functions of the Rb tumor suppressor in developing central nervous system. AB - The retinoblastoma tumor suppressor (RB) plays an important role in the regulation of cell cycle progression and terminal differentiation of many cell types. Rb(-/-) mouse embryos die at midgestation with defects in cell cycle regulation, control of apoptosis and terminal differentiation. However, chimeric mice composed of wild-type and Rb-deficient cells are viable and show minor abnormalities. To determine the role of Rb in development more precisely, we analyzed chimeric embryos and adults made with marked Rb(-/-) cells. Like their germline Rb(-/-) counterparts, brains of midgestation chimeric embryos exhibited extensive ectopic S-phase entry. In Rb-mutants, this is accompanied by widespread apoptosis. However, in chimeras, the majority of Rb-deficient cells survived and differentiated into neuronal fates. Rescue of Rb(-/-) neurons in the presence of wild-type cells occurred after induction of the p53 pathway and led to accumulation of cells with 4n DNA content. Therefore, the role of Rb during development can be divided into a cell-autonomous function in exit from the cell cycle and a non-cell-autonomous role in the suppression of apoptosis and induction of differentiation. PMID- 11432829 TI - PTP-PEST, a scaffold protein tyrosine phosphatase, negatively regulates lymphocyte activation by targeting a unique set of substrates. AB - There is increasing interest in elucidating the mechanisms involved in the negative regulation of lymphocyte activation. Herein, we show that the cytosolic protein tyrosine phosphatase PTP-PEST is expressed abundantly in a wide variety of haemopoietic cell types, including B cells and T cells. In a model B-cell line, PTP-PEST was found to be constitutively associated with several signalling molecules, including Shc, paxillin, Csk and Cas. The interaction between Shc and PTP-PEST was augmented further by antigen receptor stimulation. Overexpression studies, antisense experiments and structure-function analyses provided evidence that PTP-PEST is an efficient negative regulator of lymphocyte activation. This function correlated with the ability of PTP-PEST to induce dephosphorylation of Shc, Pyk2, Fak and Cas, and inactivate the Ras pathway. Taken together, these data suggest that PTP-PEST is a novel and unique component of the inhibitory signalling machinery in lymphocytes. PMID- 11432830 TI - Notch signaling is a direct determinant of keratinocyte growth arrest and entry into differentiation. AB - The role of Notch signaling in growth/differentiation control of mammalian epithelial cells is still poorly defined. We show that keratinocyte-specific deletion of the Notch1 gene results in marked epidermal hyperplasia and deregulated expression of multiple differentiation markers. In differentiating primary keratinocytes in vitro endogenous Notch1 is required for induction of p21WAF1/Cip1 expression, and activated Notch1 causes growth suppression by inducing p21WAF1/Cip1 expression. Activated Notch1 also induces expression of 'early' differentiation markers, while suppressing the late markers. Induction of p21WAF1/Cip1 expression and early differentiation markers occur through two different mechanisms. The RBP-Jkappa protein binds directly to the endogenous p21 promoter and p21 expression is induced specifically by activated Notch1 through RBP-Jkappa-dependent transcription. Expression of early differentiation markers is RBP-Jkappa-independent and can be induced by both activated Notch1 and Notch2, as well as the highly conserved ankyrin repeat domain of the Notch1 cytoplasmic region. Thus, Notch signaling triggers two distinct pathways leading to keratinocyte growth arrest and differentiation. PMID- 11432831 TI - A direct interaction between JNK1 and CrkII is critical for Rac1-induced JNK activation. AB - CrkII, a cellular homolog of v-crk, belongs to a family of adaptor proteins that play a central role in signal transduction cascades. We demonstrate that CrkII interacts directly with c-Jun N-terminal kinase 1 (JNK1). A proline-rich sequence of JNK1 is critical for the interaction of the kinase with the N-terminal Src homology 3 (SH3) domain of CrkII. JNK1 is localized with CrkII in membrane ruffles of Crk-overexpressing cells in a Rac1-dependent manner. A JNK1 mutant (K340A) that fails to interact with CrkII is defective in Rac/epidermal growth factor-induced activation, but remains responsive to UVC irradiation. Furthermore, CrkII recruits JNK1 to a p130Cas multiprotein complex where it may be activated through a hematopoietic progenitor kinase 1- and mitogen-activated protein kinase kinase 4-dependent pathway. Together, the results presented here argue for a new mechanism of regulation of the JNK pathway through the CrkII p130Cas adaptor complex. PMID- 11432832 TI - Mixed lineage kinase-dependent JNK activation is governed by interactions of scaffold protein JIP with MAPK module components. AB - It has been proposed that JNK-interacting proteins (JIP) facilitate mixed lineage kinase-dependent signal transduction to JNK by aggregating the three components of a JNK module. A new model for the assembly and regulation of these modules is proposed based on several observations. First, artificially induced dimerization of dual leucine zipper-bearing kinase (DLK) confirmed that DLK dimerization is sufficient to induce DLK activation. Secondly, under basal conditions, DLK associated with JIP is held in a monomeric, unphosphorylated and catalytically inactive state. Thirdly, JNK recruitment to JIP coincided with significantly decreased affinity of JIP and DLK. JNK promoted the dimerization, phosphorylation and activation of JIP-associated DLK. Similarly, treatment of cells with okadaic acid inhibited DLK association with JIP and resulted in DLK dimerization in the presence of JIP. In summary, JIP maintains DLK in a monomeric, unphosphorylated, inactive state. Upon stimulation, JNK-JIP binding affinity increases while JIP DLK interaction affinity is attenuated. Dissociation of DLK from JIP results in subsequent DLK dimerization, autophosphorylation and module activation. Evidence is provided that this model holds for other MLK-dependent JNK modules. PMID- 11432833 TI - Pin1 is overexpressed in breast cancer and cooperates with Ras signaling in increasing the transcriptional activity of c-Jun towards cyclin D1. AB - Phosphorylation on serines or threonines preceding proline (Ser/Thr-Pro) is a major signaling mechanism. The conformation of a subset of phosphorylated Ser/Thr Pro motifs is regulated by the prolyl isomerase Pin1. Inhibition of Pin1 induces apoptosis and may also contribute to neuronal death in Alzheimer's disease. However, little is known about the role of Pin1 in cancer or in modulating transcription factor activity. Here we report that Pin1 is strikingly overexpressed in human breast cancers, and that its levels correlate with cyclin D1 levels in tumors. Overexpression of Pin1 increases cellular cyclin D1 protein and activates its promoter. Furthermore, Pin1 binds c-Jun that is phosphorylated on Ser63/73-Pro motifs by activated JNK or oncogenic Ras. Moreover, Pin1 cooperates with either activated Ras or JNK to increase transcriptional activity of c-Jun towards the cyclin D1 promoter. Thus, Pin1 is up-regulated in human tumors and cooperates with Ras signaling in increasing c-Jun transcriptional activity towards cyclin D1. Given the crucial roles of Ras signaling and cyclin D1 overexpression in oncogenesis, our results suggest that overexpression of Pin1 may promote tumor growth. PMID- 11432834 TI - The eukaryotic response regulator Skn7p regulates calcineurin signaling through stabilization of Crz1p. AB - To survive ionic, pH and pheromone stress, the yeast Saccharomyces cerevisiae activates signaling through the Ca2+-activated phosphatase calcineurin to the transcription factor Crz1p/Tcn1p. We show that the overexpression of SKN7, a response-regulator transcription factor, activates transcription from a calcineurin/Crz1p-dependent response element (CDRE). Ca2+-induced, calcineurin/Crz1p-dependent activation of several genes is reduced in skn7 mutants. Skn7p modulates CDRE-dependent transcription by affecting Crz1p protein levels. Specifically, the rate of Crz1p turnover is increased in skn7 mutants. Calcineurin, but not its phosphatase activity, is required for Skn7p-mediated Crz1p stabilization. Skn7p binds to both calcineurin and Crz1p in vitro, and we suggest that this interaction is required for Skn7p regulation of Crz1p. The DNA binding and internal coiled-coil domains, but not the response- regulator phosphorylation of Skn7p, are necessary for Crz1p-dependent transcriptional activation and Crz1p stabilization by Skn7 in vivo. The DNA-binding domain of Skn7p is also required for binding to Crz1p and calcineurin in vitro. Thus, we propose that Skn7p protects Crz1p from degradation by binding to it and calcineurin through its DNA-binding domain. PMID- 11432835 TI - Rsk2 allosterically activates estrogen receptor alpha by docking to the hormone binding domain. AB - We describe a novel mechanism for transcriptional regulation, in which docking of p90 ribosomal S6 kinase 2 (Rsk2) to the hormone-binding domain (HBD) of estrogen receptor alpha (ERalpha) induces a conformational change that enhances the transcriptional activation function contained in the HBD. A constitutively active mutant of Rsk2 specifically enhances ERalpha-mediated transcription by phosphorylation of Ser167 in ERalpha and by physically associating with residues 326-394 of the ERalpha HBD. The anti-estrogen 4-hydroxytamoxifen blocks Rsk2 mediated activation of ERalpha, by inducing a conformation of ERalpha in which the Rsk2 docking site is masked. Transcriptional activation and docking are specific for ERalpha and do not occur with the related isoform, ERbeta. ERalpha phosphorylation, docking and transcriptional activation are regulated by the Rsk2 N-terminal kinase domain. The allosteric regulation of a target protein, independent of phosphorylation, may be paradigmatic of a general function for protein kinase docking sites. PMID- 11432836 TI - PML mediates the interferon-induced antiviral state against a complex retrovirus via its association with the viral transactivator. AB - The promyelocytic leukaemia (PML) protein localizes in the nucleus both in the nucleoplasm and in matrix-associated multiprotein complexes known as nuclear bodies (NBs). The number and the intensity of PML NBs increase in response to interferon (IFN). Overexpression of PML affects the replication of vesicular stomatitis virus and influenza virus. However, PML has a less powerful antiviral activity against these viruses than the IFN mediator MxA. Here, we show that overexpression of PML, but not that of Mx1 or MxA, leads to a drastic decrease of a complex retrovirus, the human foamy virus (HFV), gene expression. PML represses HFV transcription by complexing the HFV transactivator, Tas, preventing its direct binding to viral DNA. This physical interaction requires the N-terminal region of Tas and the RING finger of PML, but does not necessitate PML localization in NBs. Finally, we show that IFN treatment inhibits HFV replication in wild-type but not in PML-/- cells. These findings point to a role for PML in transcriptional repression and suggest that PML could play a key role in mediating an IFN-induced antiviral state against a complex retrovirus. PMID- 11432837 TI - Spt16-Pob3 and the HMG protein Nhp6 combine to form the nucleosome-binding factor SPN. AB - Yeast Spt16/Cdc68 and Pob3 form a heterodimer that acts in both DNA replication and transcription. This is supported by studies of new alleles of SPT16 described here. We show that Spt16-Pob3 enhances HO transcription through a mechanism that is affected by chromatin modification, since some of the defects caused by mutations can be suppressed by deleting the histone deacetylase Rpd3. While otherwise conserved among many eukaryotes, Pob3 lacks the HMG1 DNA-binding motif found in similar proteins such as the SSRP1 subunit of human FACT. SPT16 and POB3 display strong genetic interactions with NHP6A/B, which encodes an HMG1 motif, suggesting that these gene products function coordinately in vivo. While Spt16 Pob3 and Nhp6 do not appear to form stable heterotrimers, Nhp6 binds to nucleosomes and these Nhp6-nucleosomes can recruit Spt16-Pob3 to form SPN nucleosomes. These complexes have altered electrophoretic mobility and a distinct pattern of enhanced sensitivity to DNase I. These results suggest that Spt16-Pob3 and Nhp6 cooperate to function as a novel nucleosome reorganizing factor. PMID- 11432838 TI - An upstream repressor element plays a role in Igf2 imprinting. AB - The imprinted Igf2 gene is associated with a small upstream region that is differentially methylated on the active paternal allele. We have identified a repressor element within this sequence and shown that repression is probably mediated through a trans- acting factor, GCF2. DNA methylation of this site abrogates both protein binding and repressor activity. Targeting experiments demonstrate that this element plays a role in the repression of the maternal Igf2 gene in vivo. PMID- 11432839 TI - Human telomerase contains two cooperating telomerase RNA molecules. AB - Telomerase uses a short stretch of its intrinsic RNA molecule as template for telomere repeat synthesis. Reverse transcription of the RNA template is catalyzed by the telomerase reverse transcriptase (TERT) protein subunit. We demonstrate that human telomerase reconstituted from recombinant TERT and telomerase RNA runs as a dimer on a gel filtration column and that it contains two telomerase RNA molecules. Significantly, a telomerase heterodimer reconstituted from wild-type and mutant telomerase RNA is barely active when compared with the wild-type homodimer. We conclude that the telomerase RNA templates in the active enzyme are interdependent and functionally cooperate with each other. We discuss models that may explain the biological and enzymatic roles of telomerase dimerization. PMID- 11432840 TI - Loss of poly(ADP-ribose) polymerase-1 causes increased tumour latency in p53 deficient mice. AB - PARP-1-deficient mice display a severe defect in the base excision repair pathway leading to radiosensitivity and genomic instability. They are protected against necrosis induced by massive oxidative stress in various inflammatory processes. Mice lacking p53 are highly predisposed to malignancy resulting from defective cell cycle checkpoints, resistance to DNA damage-induced apoptosis as well as from upregulation of the iNOS gene resulting in chronic oxidative stress. Here, we report the generation of doubly null mutant mice. We found that tumour-free survival of parp-1(-/-)p53(-/-) mice increased by 50% compared with that of parp- 1(+/+)p53(-/-) mice. Tumour formation in nude mice injected with oncogenic parp 1(-/-)p53(-/-) fibroblasts was significantly delayed compared with parp 1(+/+)p53(-/-) cells. Upon gamma-irradiation, a partial restoration of S-phase radiosensitivity was found in parp-1(-/-)p53(-/-) primary fibroblasts compared with parp-1(+/+)p53(-/-) cells. In addition, iNOS expression and nitrite release were dramatically reduced in the parp-1(-/-)p53(-/-) mice compared with parp 1(+/+)p53(-/-) mice. The abrogation of the oxydated status of p53(-/-) cells, due to the absence of parp-1, may be the cause of the delay in the onset of tumorigenesis in parp-1(-/-)p53(-/-) mice. PMID- 11432841 TI - The ribonucleotide reductase inhibitor Sml1 is a new target of the Mec1/Rad53 kinase cascade during growth and in response to DNA damage. AB - The evolutionarily conserved protein kinases Mec1 and Rad53 are required for checkpoint response and growth. Here we show that their role in growth is to remove the ribonucleotide reductase inhibitor Sml1 to ensure DNA replication. Sml1 protein levels fluctuate during the cell cycle, being lowest during S phase. The disappearance of Sml1 protein in S phase is due to post-transcriptional regulation and is associated with protein phosphorylation. Both phosphorylation and diminution of Sml1 require MEC1 and RAD53. More over, failure to remove Sml1 in mec1 and rad53 mutants results in incomplete DNA replication, defective mitochondrial DNA propagation, decreased dNTP levels and cell death. Interestingly, similar regulation of Sml1 also occurs after DNA damage. In this case, the regulation requires MEC1 and RAD53, as well as other checkpoint genes. Therefore, Sml1 is a new target of the DNA damage checkpoint and its removal is a conserved function of Mec1 and Rad53 during growth and after damage. PMID- 11432842 TI - Double-check probing of DNA bending and unwinding by XPA-RPA: an architectural function in DNA repair. AB - The multiprotein factor composed of XPA and replication protein A (RPA) is an essential subunit of the mammalian nucleotide excision repair system. Although XPA-RPA has been implicated in damage recognition, its activity in the DNA repair pathway remains controversial. By replacing DNA adducts with mispaired bases or non-hybridizing analogues, we found that the weak preference of XPA and RPA for damaged substrates is entirely mediated by indirect readout of DNA helix conformations. Further screening with artificially distorted substrates revealed that XPA binds most efficiently to rigidly bent duplexes but not to single stranded DNA. Conversely, RPA recognizes single-stranded sites but not backbone bending. Thus, the association of XPA with RPA generates a double-check sensor that detects, simultaneously, backbone and base pair distortion of DNA. The affinity of XPA for sharply bent duplexes, characteristic of architectural proteins, is not compatible with a direct function during recognition of nucleotide lesions. Instead, XPA in conjunction with RPA may constitute a regulatory factor that monitors DNA bending and unwinding to verify the damage specific localization of repair complexes or control their correct three dimensional assembly. PMID- 11432843 TI - Human immunodeficiency virus type 1 integrase: arrangement of protein domains in active cDNA complexes. AB - Early steps of retroviral replication involve reverse transcription of the viral RNA genome and integration of the resulting cDNA copy into a chromosome of the host cell. The viral-encoded integrase protein carries out the initial DNA breaking and joining reactions that mediate integration. The organization of the active integrase-DNA complex is unknown, though integrase is known to act as a multimer, and high resolution structures of the isolated integrase domains have been determined. Here we use site-specific cross-linking based on disulfide bond formation to map integrase-DNA contacts in active complexes. We establish that the DNA-binding C-terminal domain of one integrase monomer acts with the central catalytic domain from another monomer at each viral cDNA end. These data allow detailed modeling of an integrase tetramer in which pairs of trans interactions link integrase dimers bound to substrate DNA. We also detected a conformational change in integrase- DNA complexes accompanying cleavage of the viral cDNA terminus. PMID- 11432844 TI - Proximity of the invariant loop of U5 snRNA to the second intron residue during pre-mRNA splicing. AB - A photoactivatable azidophenacyl group has been introduced into seven positions in the backbone of the 11 nucleotide invariant loop of U5 snRNA. By reconstituting depleted splicing extracts with reassembled U5 snRNP particles, molecular neighbors were assessed as a function of splicing. All cross-links to the pre-mRNA mapped to the second nucleotide downstream of the 5' splice site, and formed most readily when the reactive group was at the phosphate between U5 positions 42 and 43 or 43 and 44. Both their kinetics of appearance and sensitivity to oligonucleotide inhibition suggest that these cross-links capture a late state in spliceosome assembly occurring immediately prior to the first step. A later forming, second cross-linked species is a splicing product of the first cross-link, suggesting that the U5 loop backbone maintains this position through the first step. The proximity of the U5 loop backbone to the intron's 5' end provides sufficient restrictions to develop a three-dimensional model for the arrangement of RNA components in the spliceosome during the first step of pre mRNA splicing. PMID- 11432845 TI - Expression of the glucose transporter gene, ptsG, is regulated at the mRNA degradation step in response to glycolytic flux in Escherichia coli. AB - We report a novel post-transcriptional control of the ptsG gene encoding the major glucose transporter IICB(Glc). We demonstrate that the level of IICB(Glc) is markedly reduced when the glycolytic pathway is blocked by a mutation in either the pgi or pfkA gene encoding phosphoglucose isomerase or phosphofructokinase, respectively. This down-regulation of ptsG is not exerted at the transcriptional level. Both northern blot and S1 analyses demonstrate that the mutation dramatically accelerates the degradation of ptsG mRNA. The degradation of ptsG mRNA occurs in wild-type cells when alpha-methylglucoside, a non- metabolizable analog of glucose, is present in the medium. The addition of any one of the glycolytic intermediates downstream of the block prevents the degradation of ptsG mRNA. The rapid degradation of ptsG mRNA is eliminated when RNase E is thermally inactivated. We conclude that the glycolytic pathway controls ptsG expression by modulating RNase E-mediated mRNA degradation. This is the first instance in which the glycolytic flux has been shown to affect the expression of a specific gene through mRNA stability. PMID- 11432846 TI - VIP1, an Arabidopsis protein that interacts with Agrobacterium VirE2, is involved in VirE2 nuclear import and Agrobacterium infectivity. AB - T-DNA nuclear import is a central event in genetic transformation of plant cells by Agrobacterium. This event is thought to be mediated by two bacterial proteins, VirD2 and VirE2, which are associated with the transported T-DNA molecule. While VirD2 is imported into the nuclei of plant, animal and yeast cells, nuclear uptake of VirE2 occurs most efficiently in plant cells. To understand better the mechanism of VirE2 action, a cellular interactor of VirE2 was identified and its encoding gene cloned from Arabidopsis. The identified plant protein, designated VIP1, specifically bound VirE2 and allowed its nuclear import in non-plant systems. In plants, VIP1 was required for VirE2 nuclear import and Agrobacterium tumorigenicity, participating in early stages of T-DNA expression. PMID- 11432847 TI - Phosphatidylinositol 3-kinase-dependent extracellular calcium influx is essential for CX(3)CR1-mediated activation of the mitogen-activated protein kinase cascade. AB - Fractalkine, the first member of the CX(3)C chemokine family, induces leukocyte chemotaxis through activation of its high affinity receptor, CX(3)CR1. Like other chemokine receptors, CX(3)CR1 is coupled to a pertussis toxin-sensitive heterotrimeric G(i) protein, which is necessary for rapid rise in the concentration of intracellular calcium. Using a Chinese hamster ovary cell line stably transfected with the CX(3)CR1 receptor, we show that the source of calcium mobilized by fractalkine stimulation is the extracellular pool. Calcium influx is blocked by extracellular calcium chelators, as well as by divalent heavy metals such as Ni(2+), Co(2+), and Cd(2+) without affecting the integrity of intracellular stores. Remarkably, selective phosphoinositide 3-kinase (PI3K) inhibitors, wortmannin and LY294002, abolish the wave extracellular calcium, suggesting that an active PI3K is necessary for this event. The influx of extracellular calcium is in turn required to trigger the activation of the p42/44 mitogen-activated protein/extracellular signal-regulated kinase pathway, but is not necessary for other signals downstream to PI3K, such as phosphorylation of Akt. The potential role of this signaling cascade in fractalkine-mediated chemotaxis is discussed. PMID- 11432848 TI - Examining the in vivo role of the amino terminus of the essential myosin light chain. AB - The functional significance of the actin binding region at the amino terminus of the cardiac essential myosin light chain (ELC) remains obscure. Previous experiments carried out in vitro indicated that modulation of residues 5-14 could induce an inotropic effect, increasing maximal ATPase activity at submaximal Ca(2+) concentrations (Rarick, H. M., Opgenorth, T. J., von Geldern, T. W., Wu Wong, J. R., and Solaro, R. J. (1996) J. Biol. Chem. 271, 27039-27043). Using transgenesis, we effected a cardiac-specific replacement of ELC with a protein containing a 10-amino acid deletion at positions 5-14. Both the ventricular (ELC1vDelta5-14) and atrial (ELC1aDelta5-14) isoforms lacking this peptide were stably incorporated into the sarcomere at high efficiencies. Surprisingly when the kinetics of skinned fibers isolated from the ELC1vDelta5-14 or ELC1aDelta5-14 mice were examined, no alterations in either unloaded shortening or maximum shortening velocities were apparent. Myofibrillar Mg(2+)-ATPase activity was also unchanged in these preparations. No significant changes in the fiber kinetics in the cognate compartments were observed when either deletion-containing protein replaced endogenous ELC1v or ELC1a. The data indicate that the previously postulated importance of this region in mediating critical protein interactions between the cardiac ELCs and the carboxyl-terminal residues of actin in vivo should be reassessed. PMID- 11432849 TI - The first proline of PALP motif at the C terminus of presenilins is obligatory for stabilization, complex formation, and gamma-secretase activities of presenilins. AB - Mutations in presenilin (PS) genes cause early-onset familial Alzheimer's disease by increasing production of the amyloidogenic form of amyloid beta peptides ending at residue 42 (Abeta42). PS is an evolutionarily conserved multipass transmembrane protein, and all known PS proteins contain a proline-alanine leucine-proline (PALP) motif starting at proline (P) 414 (amino acid numbering based on human PS2) at the C terminus. Furthermore, missense mutations that replace the first proline of PALP with leucine (P414L) lead to a loss-of-function of PS in Drosophila melanogaster and Caenorhabditis elegans. To elucidate the roles of the PALP motif in PS structure and function, we analyzed neuro2a as well as PS1/2 null fibroblast cell lines transfected with human PS harboring mutations at the PALP motif. P414L mutation in PS2 (and its equivalent in PS1) abrogated stabilization, high molecular weight complex formation, and entry to Golgi/trans Golgi network of PS proteins, resulting in failure of Abeta42 overproduction on familial Alzheimer's disease mutant basis as well as of site-3 cleavage of Notch. These data suggest that the first proline of the PALP motif plays a crucial role in the stabilization and formation of the high molecular weight complex of PS, the latter being the active form with intramembrane proteolytic activities. PMID- 11432850 TI - Essential requirement of cytosolic phospholipase A(2) for stimulation of NADPH oxidase-associated diaphorase activity in granulocyte-like cells. AB - We have previously established a model of cytosolic phospholipase A(2) (cPLA(2)) deficient differentiated PLB-985 cells (PLB-D cells) and demonstrated that cPLA(2)-generated arachidonic acid (AA) is essential for NADPH oxidase activation. In this study we used this model to investigate the physiological role of cPLA(2) in regulation of NADPH oxidase-associated diaphorase activity. A novel diaphorase activity assay, using 4-iodonitrotetrazolium violet as an electron acceptor, was used in permeabilized neutrophils and PLB-985 cells differentiated toward the granulocytic or monocytic phenotypes. Phorbol 12 myristate 13-acetate, guanosine 5'-3-O- (thio)triphosphate (GTP gamma S), or FMLP stimulated a similar diphenylene iodonium-sensitive diaphorase activity pattern in neutrophils and in differentiated parent PLB-985 cells. This diaphorase activity was not detected in undifferentiated cells, but developed during differentiation. Furthermore, diaphorase activity could not be stimulated in permeabilized neutrophils from X-linked CGD patients and in differentiated gp91(phox)-targeted PLB-985 cells that lacked normal expression of gp91(phox), but was restored to these cells following transduction with retrovirus encoding gp91(phox). The differentiated PLB-D cells showed no diaphorase activity when stimulated by either GTP gamma S or FMLP, and only partial activation when stimulated with phorbol 12-myristate 13-acetate. Diaphorase activity in response to either agonists was fully restored by the addition of 10 microm free AA. The permeabilized cell 4-iodonitrotetrazolium violet reduction assay offers a unique tool for the evaluation of NADPH oxidase-associated diaphorase activity in stimulated whole cells. These results establish an essential and specific physiological requirement of cPLA(2)-generated AA in activation of electron transfer through the FAD reduction center of NADPH oxidase. PMID- 11432851 TI - An Abd-B class HOX.PBX recognition sequence is required for expression from the mouse Ren-1c gene. AB - Expression from the mouse Ren-1(c) gene in As4.1 cells is dependent on a proximal promoter element (PPE) located at approximately -60 and a 241-base pair enhancer region located at -2625 relative to the transcription start site. The PPE (TAATAAATCAA) is identical to a consensus HOX.PBX binding sequence. Further, PBX1b has been shown to be a component of a PPE-specific binding complex present in nuclear extracts from As4.1 cells. The binding affinities of different paralog HOX members to the PPE were examined in the absence or presence of PBX1b. HOXB6, B7, and -C8 failed to bind the PPE alone but showed weak affinity in the presence of PBX1b. In contrast, HOXD10 and to a lesser degree HOXB9 bound the PPE with high affinities regardless of whether PBX1b was present. Abd-B HOX members, including HOXD10, -A10, -A9, -B9, and -C9, are expressed in As4.1 cells. The ability of HOX and PBX1b to form a ternary complex with PREP1 on the PPE is also demonstrated both in vivo and in vitro. Point mutations in either the HOX or PBX half-site of the PPE disrupted the formation of the HOX.PBX complex and dramatically decreased transcriptional activity of the Ren-1(c) gene demonstrating that both the HOX and PBX half-sites are critical for mouse renin gene expression. These results strongly implicate Abd-B class Hox genes and their cofactors as major determinants of the sites of renin expression. PMID- 11432852 TI - Identification of a critical Sp1 site within the endoglin promoter and its involvement in the transforming growth factor-beta stimulation. AB - Endoglin, a component of the transforming growth factor-beta (TGF-beta) receptor complex expressed on endothelial cells, is involved in cardiovascular morphogenesis and vascular remodeling, as exemplified by the fact that the endoglin gene is the target for the autosomal dominant disorder known as hereditary hemorrhagic telangiectasia type 1. Since haploinsufficiency is the underlying mechanism for hereditary hemorrhagic telangiectasia type 1, understanding the regulation of endoglin gene expression appears to be a crucial step to correct the disease. In this study we have identified an Sp1 site at -37 as a critical element for the basal transcription of the endoglin TATA-less promoter. Since endoglin promoter activity is stimulated by TGF-beta and this stimulation is located at the Sp1-containing proximal region, we have investigated the possible involvement of Sp1 in the TGF-beta-mediated induction. Mutation of the Sp1-binding sequence, or addition of the Sp1 inhibitor WP631, abolished both the basal transcription activity and the TGF-beta responsiveness of the endoglin promoter. Binding of Sp1 and Smad3 to the proximal promoter region -50/-29 was evidenced by electrophoretic mobility shift assays and DNA affinity precipitation studies. Furthermore, synergistic cooperation on the promoter activity between Sp1 and TGF-beta or Smad3 could be demonstrated by co transfection experiments of reporter promoter constructs. The molecular mechanism underlying this cooperation appears to involve a direct physical interaction between Sp1 and Smad3/Smad4. PMID- 11432853 TI - ATP utilization by yeast replication factor C. I. ATP-mediated interaction with DNA and with proliferating cell nuclear antigen. AB - Eukaryotic replication factor C is the heteropentameric complex that loads the replication clamp proliferating cell nuclear antigen (PCNA) onto primed DNA. In this study we used a derivative, designated RFC, with a N-terminal truncation of the Rfc1 subunit removing a DNA-binding domain not required for clamp loading. Interactions of yeast RFC with PCNA and DNA were studied by surface plasmon resonance. Binding of RFC to PCNA was stimulated by either adenosine (3 thiotriphosphate) (ATPgammaS) or ATP. RFC bound only to primer-template DNA coated with the single-stranded DNA-binding protein RPA if ATPgammaS was also present. Binding occurred without dissociation of RPA. ATP did not stimulate binding of RFC to DNA, suggesting that hydrolysis of ATP dissociated DNA-bound RFC. However, when RFC and PCNA together were flowed across the DNA chip in the presence of ATP, a signal was observed suggesting loading of PCNA by RFC. With ATPgammaS present instead of ATP, long-lived response signals were observed indicative of loading complexes arrested on the DNA. A primer with a 3' single stranded extension also allowed loading of PCNA; yet turnover of the reaction intermediates was dramatically slowed down. Filter binding experiments and analysis of proteins bound to DNA-magnetic beads confirmed the conclusions drawn from the surface plasmon resonance studies. PMID- 11432854 TI - ATP utilization by yeast replication factor C. III. The ATP-binding domains of Rfc2, Rfc3, and Rfc4 are essential for DNA recognition and clamp loading. AB - The conserved lysine in the Walker A motif of the ATP-binding domain encoded by the yeast RFC1, RFC2, RFC3, and RFC4 genes was mutated to glutamic acid. Complexes of replication factor C with a N-terminal truncation (Delta2-273) of the Rfc1 subunit (RFC) containing a single mutant subunit were overproduced in Escherichia coli for biochemical analysis. All of the mutant RFC complexes were capable of interacting with PCNA. Complexes containing a rfc1-K359E mutation were similar to wild type in replication activity and ATPase activity; however, the mutant complex showed increased susceptibility to proteolysis. In contrast, complexes containing either a rfc2-K71E mutation or a rfc3-K59E mutation were severely impaired in ATPase and clamp loading activity. In addition to their defects in ATP hydrolysis, these complexes were defective for DNA binding. A mutant complex containing the rfc4-K55E mutation performed as well as a wild type complex in clamp loading, but only at very high ATP concentrations. Mutant RFC complexes containing rfc2-K71R or rfc3-K59R, carrying a conservative lysine --> arginine mutation, had much milder clamp loading defects that could be partially (rfc2-K71R) or completely (rfc3-K59R) suppressed at high ATP concentrations. PMID- 11432856 TI - ATP utilization by yeast replication factor C. II. Multiple stepwise ATP binding events are required to load proliferating cell nuclear antigen onto primed DNA. AB - Binding of adenosine (3-thiotriphosphate) (ATPgammaS), a nonhydrolyzable analog of ATP, to replication factor C with a N-terminal truncation (Delta2-273) of the Rfc1 subunit (RFC) was studied by filter binding. RFC alone bound 1.8 ATPgammaS molecules. However, when either PCNA or primer-template DNA were also present 2.6 or 2.7 ATPgammaS molecules, respectively, were bound. When both PCNA and DNA were present 3.6 ATPgammaS molecules were bound per RFC. Order of addition experiments using surface plasmon resonance indicate that RFC forms an ATP-mediated binary complex with PCNA prior to formation of a ternary DNA.PCNA.RFC complex. An ATP mediated complex between RFC and DNA was not competent for binding PCNA, and the RFC.DNA complex dissociated with hydrolysis of ATP. Based on these experiments a model is proposed in which: (i) RFC binds two ATPs (RFC.ATP(2)); (ii) this complex binds PCNA (PCNA.RFC.ATP(2)), which goes through a conformational change to reveal a binding site for one additional ATP (PCNA.RFC.ATP(3)); (iii) this complex can bind DNA to yield DNA.PCNA.RFC.ATP(3); (iv) a conformational change in the latter complex reveals a fourth binding site for ATP; and (v) the DNA.PCNA.RFC.ATP(4) complex is finally competent for completion of PCNA loading and release of RFC upon hydrolysis of ATP. PMID- 11432857 TI - The delta and delta ' subunits of the DNA polymerase III holoenzyme are essential for initiation complex formation and processive elongation. AB - delta and delta' are required for assembly of the processivity factor beta(2) onto primed DNA in the DNA polymerase III holoenzyme-catalyzed reaction. We developed protocols for generating highly purified preparations of delta and delta'. In holoenzyme reconstitution assays, delta' could not be replaced by delta, tau, or gamma, even when either of the latter were present at a 10,000 fold molar excess. Likewise, delta could not be replaced by delta', tau, or gamma. Bacterial strains bearing chromosomal knockouts of either the holA(delta) or holB(delta') genes were not viable, demonstrating that both delta and delta' are essential. Western blots of isolated initiation complexes demonstrated the presence of both delta and delta'. However, in the absence of chipsi and single stranded DNA-binding protein, a stable initiation complex lacking deltadelta' was isolated by gel filtration. Lack of delta-delta' decreased the rate of elongation about 3-fold, and the extent of processive replication was significantly decreased. Adding back delta-delta' but not chipsi, delta, or delta' alone restored the diminished activity, indicating that in addition to being key components required for the beta loading activity of the DnaX complex, deltadelta' is present in initiation complex and is required for processive elongation. PMID- 11432858 TI - Molecular determinants of receptor binding and signaling by the CX3C chemokine fractalkine. AB - Fractalkine/CX3CL1 is a membrane-tethered chemokine that functions as a chemoattractant and adhesion protein by interacting with the receptor CX3CR1. To understand the molecular basis for the interaction, an extensive mutagenesis study of fractalkine's chemokine domain was undertaken. The results reveal a cluster of basic residues (Lys-8, Lys-15, Lys-37, Arg-45, and Arg-48) and one aromatic (Phe-50) that are critical for binding and/or signaling. The mutant R48A could bind but not induce chemotaxis, demonstrating that Arg-48 is a signaling trigger. This result also shows that signaling residues are not confined to chemokine N termini, as generally thought. F50A showed no detectable binding, underscoring its importance to the stability of the complex. K15A displayed unique signaling characteristics, eliciting a wild-type calcium flux but minimal chemotaxis, suggesting that this mutant can activate some, but not all, pathways required for migration. Fractalkine also binds the human cytomegalovirus receptor US28, and analysis of the mutants indicates that US28 recognizes many of the same epitopes of fractalkine as CX3CR1. Comparison of the binding surfaces of fractalkine and the CC chemokine MCP-1 reveals structural details that may account for their dual recognition by US28 and their selective recognition by host receptors. PMID- 11432859 TI - Cop, a caspase recruitment domain-containing protein and inhibitor of caspase-1 activation processing. AB - The production of bio-active interleukin-1beta (IL-1beta), a pro-inflammatory cytokine, is mediated by activated caspase-1. One of the known molecular mechanisms underlying pro-caspase-1 processing and activation involves binding of the caspase-1 prodomain to a caspase recruitment domain (CARD)-containing serine/threonine kinase known as RIP2/CARDIAK/RICK. We have identified a novel protein, COP (CARD only protein), which has a high degree of sequence identity to the caspase-1 prodomain. COP binds to both RIP2 and the caspase-1 prodomain and inhibits RIP2-induced caspase-1 oligomerization. COP inhibits caspase- 1-induced IL-1beta secretion as well as lipopolysaccharide-induced IL-1beta secretion in transfected cells. Our data indicate that COP can regulate IL-1beta secretion, implying that COP may play a role in down-regulating inflammatory responses analogous to the CARD protein ICEBERG. PMID- 11432860 TI - Arg(1098) is critical for the chloride dependence of human angiotensin I converting enzyme C-domain catalytic activity. AB - Angiotensin (Ang) I-converting enzyme (ACE) is a Zn(2+) metalloprotease with two homologous catalytic domains. Both the N- and C-terminal domains are peptidyl dipeptidases. Hydrolysis by ACE of its decapeptide substrate Ang I is increased by Cl(-), but the molecular mechanism of this regulation is unclear. A search for single substitutions to Gln among all conserved basic residues (Lys/Arg) in human ACE C-domain identified R1098Q as the sole mutant that lacked Cl(-) dependence. Cl(-) dependence is also lost when the equivalent Arg in the N-domain, Arg(500), is substituted with Gln. The Arg(1098) to Lys substitution reduced Cl(-) binding affinity by approximately 100-fold. In the absence of Cl(-), substrate binding affinity (1/K(m)) of and catalytic efficiency (k(cat)/K(m)) for Ang I hydrolysis are increased 6.9- and 32-fold, respectively, by the Arg(1098) to Gln substitution, and are similar (<2-fold difference) to the respective wild-type C domain catalytic constants in the presence of optimal [Cl(-)]. The Arg(1098) to Gln substitution also eliminates Cl(-) dependence for hydrolysis of tetrapeptide substrates, but activity toward these substrates is similar to that of the wild type C-domain in the absence of Cl(-). These findings indicate that: 1) Arg(1098) is a critical residue of the C-domain Cl(-)-binding site and 2) a basic side chain is necessary for Cl(-) dependence. For tetrapeptide substrates, the inability of R1098Q to recreate the high affinity state generated by the Cl(-)-C domain interaction suggests that substrate interactions with the enzyme-bound Cl( ) are much more important for the hydrolysis of short substrates than for Ang I. Since Cl(-) concentrations are saturating under physiological conditions and Arg(1098) is not critical for Ang I hydrolysis, we speculate that the evolutionary pressure for the maintenance of the Cl(-)-binding site is its ability to allow cleavage of short cognate peptide substrates at high catalytic efficiencies. PMID- 11432861 TI - Effects of phosphocreatine on apoptosis in a cell-free system. AB - The characteristic morphological and biochemical changes during caspase-mediated apoptosis can be reproduced to a large extent in a Xenopus laevis egg extract cell-free system by addition of mouse liver nuclei and exogenous cytochrome c. We show that in this system phosphocreatine accelerated the apoptotic morphological changes of the nuclei, but selectively inhibited DNA fragmentation. Western blot showed that the degradation of lamins A and C is accelerated, which is possibly responsible for the nuclear changes during cell apoptosis. However, the degradation of ICAD/DFF45-like protein in the egg extracts is inhibited in a time dependent manner. Exogenous creatine, ATP, and several organic acids have no effect on DNA fragmentation, excluding the possibility that creatine, ATP, or acidic conditions resulting from phosphocreatine are responsible for inhibiting DNA fragmentation. Lithium chloride, a kinase inhibitor, can overcome the phosphocreatine effects and can restore DNA fragmentation. Our results indicate that phosphocreatine protects ICAD/DFF45-like protein from proteolysis, probably through kinase actions, resulting in its resistance to caspase cleavage and leading to an inhibition of DNA fragmentation. PMID- 11432862 TI - The kissing hairpin sequence promotes recombination within the HIV-I 5' leader region. AB - The role of RNA-RNA template interactions in facilitating recombination during reverse transcription of minus strand DNA has been examined. The tested hypothesis is that template switching by reverse transcriptase is promoted at sites where homologous regions of two RNAs are brought in close proximity via stable intertemplate interactions. Frequency and distribution of template switching between homologous donor and acceptor RNAs were examined within the human immunodeficiency virus type I (HIV-I) 5'-untranslated region (UTR) containing the dimer initiation sequence (DIS). Results were compared with control nondimerizing templates from the pol region. The dimerizing UTR templates displayed a 4-fold higher transfer efficiency than the control. A striking 53% of transfers in the UTR mapped near the DIS, of which two-thirds occurred immediately 5' to this sequence. In the UTR template, deletion of the DIS hairpin disrupted template dimerization and caused a 4-fold drop in transfer efficiency. Insertion of the DIS within the pol template increased both dimerization and transfer efficiency. Transfer distributions revealed that in both sets of templates, DIS-induced dimerization increased the efficiency of transfers across the whole template, with the transfers peaking around the dimerization site. Overall, these results suggest that template dimerization facilitated by the unique geometry of the DIS-promoted kissing interactions effectively promotes recombination within the HIV-I 5'-UTR. PMID- 11432863 TI - Identification of another actin-related protein (Arp) 2/3 complex binding site in neural Wiskott-Aldrich syndrome protein (N-WASP) that complements actin polymerization induced by the Arp2/3 complex activating (VCA) domain of N-WASP. AB - Neural Wiskott-Aldrich syndrome protein (N-WASP) is an essential regulator of actin cytoskeleton formation via its association with the actin-related protein (Arp) 2/3 complex. It is believed that the C-terminal Arp2/3 complex-activating domain (verprolin homology, cofilin homology, and acidic (VCA) or C-terminal region of WASP family proteins domain) of N-WASP is usually kept masked (autoinhibition) but is opened upon cooperative binding of upstream regulators such as Cdc42 and phosphatidylinositol 4,5-bisphosphate (PIP2). However, the mechanisms of autoinhibition and association with Arp2/3 complex are still unclear. We focused on the acidic region of N-WASP because it is thought to interact with Arp2/3 complex and may be involved in autoinhibition. Partial deletion of acidic residues from the VCA portion alone greatly reduced actin polymerization activity, demonstrating that the acidic region contributes to Arp2/3 complex-mediated actin polymerization. Surprisingly, the same partial deletion of the acidic region in full-length N-WASP led to constitutive activity comparable with the activity seen with the VCA portion. Therefore, the acidic region in full-length N-WASP plays an indispensable role in the formation of the autoinhibited structure. This mutant contains WASP-homology (WH) 1 domain with weak affinity to the Arp2/3 complex, leading to activity in the absence of part of the acidic region. Furthermore, the actin comet formed by the DeltaWH1 mutant of N-WASP was much smaller than that of wild-type N-WASP. Partial deletion of acidic residues did not affect actin comet size, indicating the importance of the WH1 domain in actin structure formation. Collectively, the acidic region of N WASP plays an essential role in Arp2/3 complex activation as well as in the formation of the autoinhibited structure, whereas the WH1 domain complements the activation of the Arp2/3 complex achieved through the VCA portion. PMID- 11432864 TI - The mechanism of dephosphorylation of extracellular signal-regulated kinase 2 by mitogen-activated protein kinase phosphatase 3. AB - The mitogen-activated protein (MAP) kinase phosphatase-3 (MKP3) is a dual specificity phosphatase that specifically inactivates one subfamily of MAP kinases, the extracellular signal-regulated kinases (ERKs). Inactivation of MAP kinases occurs by dephosphorylation of Thr(P) and Tyr(P) in the TXY kinase activation motif. To gain insight into the mechanism of ERK2 inactivation by MKP3, we have carried out an analysis of the MKP3-catalyzed dephosphorylation of the phosphorylated ERK2. We find that ERK2/pTpY dephosphorylation by MKP3 involves an ordered, distributive mechanism in which MKP3 binds the bisphosphorylated ERK2/pTpY, dephosphorylates Tyr(P) first, dissociates and releases the monophosphorylated ERK2/pT, which is then subjected to dephosphorylation by a second MKP3, yielding the fully dephosphorylated ERK2. The bisphosphorylated ERK2 is a highly specific substrate for MKP3 with a k(cat)/K(m) of 3.8 x 10(6) m(-1) s(-1), which is more than 6 orders of magnitude higher than that for small molecule aryl phosphates and an ERK2-derived phosphopeptide encompassing the pTEpY motif. This strikingly high substrate specificity displayed by MKP3 may result from a combination of high affinity binding interactions between the N-terminal domain of MKP3 and ERK2 and specific ERK2 induced allosteric activation of the MKP3 C-terminal phosphatase domain. PMID- 11432865 TI - Suppression of endothelial cell apoptosis by high density lipoproteins (HDL) and HDL-associated lysosphingolipids. AB - Apoptotic cell death following injury of vascular endothelium is assumed to play an important role in the pathogenesis of atherosclerosis. In this report, we demonstrate that high density lipoproteins (HDL), a major anti-atherogenic lipoprotein fraction, protect endothelial cells against growth factor deprivation induced apoptosis. HDL blocked the mitochondrial pathway of apoptosis by inhibiting dissipation of mitochondrial potential (Deltapsi(m)), generation of reactive oxygen species, and release of cytochrome c into the cytoplasm. As a consequence, HDL prevented activation of caspases 9 and 3 and apoptotic alterations of the plasma membrane such as increase of permeability and translocation of phosphatidylserine. Treatment of endothelial cells with HDL induced activation of the protein kinase Akt, an ubiquitous transducer of anti apoptotic signals, and led to phosphorylation of BAD, a major Akt substrate. Suppression of Akt activity both by wortmannin and LY-294002 or by a dominant negative Akt mutant abolished the anti-apoptotic effect of HDL. Two bioactive lysosphingolipids present in HDL particles, sphingosylphosphorylcholine and lysosulfatide, fully mimicked the survival effect of HDL by blocking the mitochondrial pathway of apoptosis and potently activating Akt. In conclusion, the present study identifies HDL as a carrier of endogenous endothelial survival factors and suggests that inhibition of endothelial apoptosis by HDL-associated lysosphingolipids may represent an important and novel aspect of the anti atherogenic activity of HDL. PMID- 11432866 TI - Adenylyl cyclase 3 mediates prostaglandin E(2)-induced growth inhibition in arterial smooth muscle cells. AB - Arterial smooth muscle cell (SMC) proliferation contributes to a number of vascular pathologies. Prostaglandin E(2) (PGE(2)), produced by the endothelium and by SMCs themselves, acts as a potent SMC growth inhibitor. The growth inhibitory effects of PGE(2) are mediated through activation of G-protein-coupled membrane receptors, activation of adenylyl cyclases (ACs), formation of cAMP, and subsequent inhibition of mitogenic signal transduction pathways in SMCs. Of the 10 different mammalian AC isoforms known today, seven isoforms (AC2-7 and AC9) are expressed in SMCs from various species. We show that, despite the presence of several different AC isoforms, the principal AC isoform activated by PGE(2) in human arterial SMCs is a calmodulin kinase II-inhibited AC with characteristics similar to those of AC3. AC3 is expressed in isolated human arterial SMCs and in intact aorta. We further show that arterial SMCs isolated from AC3-deficient mice are resistant to PGE(2)-induced growth inhibition. In summary, AC3 is the principal AC isoform activated by PGE(2) in arterial SMCs, and AC3 mediates the growth-inhibitory effects of PGE(2). Because AC3 activity is inhibited by intracellular calcium through calmodulin kinase II, AC3 may serve as an important integrator of growth-inhibitory signals that stimulate cAMP formation and growth factors that increase intracellular calcium. PMID- 11432867 TI - Bovine cytosolic 5'-nucleotidase acts through the formation of an aspartate 52 phosphoenzyme intermediate. AB - Cytosolic 5'-nucleotidase/phosphotransferase (cN-II), specific for purine monophosphates and their deoxyderivatives, acts through the formation of a phosphoenzyme intermediate. Phosphate may either be released leading to 5' mononucleotide hydrolysis or be transferred to an appropriate nucleoside acceptor, giving rise to a mononucleotide interconversion. Chemical reagents specifically modifying aspartate and glutamate residues inhibit the enzyme, and this inhibition is partially prevented by cN-II substrates and physiological inhibitors. Peptide mapping experiments with the phosphoenzyme previously treated with tritiated borohydride allowed isolation of a radiolabeled peptide. Sequence analysis demonstrated that radioactivity was associated with a hydroxymethyl derivative that resulted from reduction of the Asp-52-phosphate intermediate. Site-directed mutagenesis experiments confirmed the essential role of Asp-52 in the catalytic machinery of the enzyme and suggested also that Asp-54 assists in the formation of the acyl phosphate species. From sequence alignments we conclude that cytosolic 5'-nucleotidase, along with other nucleotidases, belong to a large superfamily of hydrolases with different substrate specificities and functional roles. PMID- 11432868 TI - Adenovirus-mediated rescue of lipoprotein lipase-deficient mice. Lipolysis of triglyceride-rich lipoproteins is essential for high density lipoprotein maturation in mice. AB - Lipoprotein lipase (LPL) is the rate-limiting enzyme for the hydrolysis of triglycerides and the subsequent uptake of free fatty acids in extrahepatic tissues. Deficiency of LPL in humans (Type I hyperlipoproteinemia) is associated with massive chylomicronemia, low high density lipoprotein (HDL) cholesterol levels, and recurrent attacks of pancreatitis when not controlled by a strict diet. In contrast to humans, homozygous LPL knock-out mice (L0) do not survive suckling and die between 18 and 24 h after birth. In this study, an adenovirus based protocol was utilized for the transient expression of LPL during the suckling period in an effort to rescue L0 mice. After a single intraperitoneal injection of 5x10(9) plaque-forming units of LPL-expressing virus immediately after birth, more than 90% of L0 mice survived the first days of life. 3% of L0 mice survived the entire suckling period and lived for up to 20 months, although LPL activity in mouse tissues and postheparin plasma was undetectable in all animals after 6 weeks of age. Adult LPL-deficient mice were smaller than their littermates until 2-3 months of age and exhibited very high triglyceride levels in the fed (4997 +/- 1102 versus 113.4 +/- 18.7 mg/dl) and fasted state (2007 +/- 375 versus 65.5 +/- 7.4 mg/dl). Plasma total cholesterol levels, free fatty acids, and ketone bodies were elevated in L0 mice, whereas plasma glucose was normal. Most strikingly, L0 mice lacked apoA-I-containing prebeta-HDL particles as well as mature HDL resulting in undetectable HDL cholesterol and HDL-apoA-I levels. HDL deficiency in plasma was evident despite normal apoA-I mRNA levels in the liver and normal apoA-I protein levels in plasma, which were predominantly found in the chylomicron fraction. The absence of prebeta-HDL and mature HDL particles supports the concept that the lipolysis of triglyceride-rich lipoproteins is an essential step for HDL maturation. PMID- 11432869 TI - Molecular and functional analysis of a novel neuronal vesicular glutamate transporter. AB - Glutamate is the major excitatory neurotransmitter in the mammalian central nervous system. Packaging and storage of glutamate into glutamatergic neuronal vesicles requires ATP-dependent vesicular glutamate uptake systems, which utilize the electrochemical proton gradient as a driving force. VGLUT1, the first identified vesicular glutamate transporter, is only expressed in a subset of glutamatergic neurons. We report here the molecular cloning and functional characterization of a novel glutamate transporter, VGLUT2, from mouse brain. VGLUT2 has all major functional characteristics of a synaptic vesicle glutamate transporter, including ATP dependence, chloride stimulation, substrate specificity, and substrate affinity. It has 75 and 79% amino acid identity with human and rat VGLUT1, respectively. However, expression patterns of VGLUT2 in brain are different from that of VGLUT1. In addition, VGLUT2 activity is dependent on both membrane potential and pH gradient of the electrochemical proton gradient, whereas VGLUT1 is primarily dependent on only membrane potential. The presence of VGLUT2 in brain regions lacking VGLUT1 suggests that the two isoforms together play an important role in vesicular glutamate transport in glutamatergic neurons. PMID- 11432870 TI - Effect of the structure of natural sterols and sphingolipids on the formation of ordered sphingolipid/sterol domains (rafts). Comparison of cholesterol to plant, fungal, and disease-associated sterols and comparison of sphingomyelin, cerebrosides, and ceramide. AB - Ordered lipid domains enriched in sphingolipids and cholesterol (lipid rafts) have been implicated in numerous functions in biological membranes. We recently found that lipid domain/raft formation is dependent on the sterol component having a structure that allows tight packing with lipids having saturated acyl chains (Xu, X., and London, E. (2000) Biochemistry 39, 844-849). In this study, the domain-promoting activities of various natural sterols were compared with that of cholesterol using both fluorescence quenching and detergent insolubility methods. Using model membranes, it was shown that, like cholesterol, both plant and fungal sterols promote the formation of tightly packed, ordered lipid domains by lipids with saturated acyl chains. Surprisingly ergosterol, a fungal sterol, and 7-dehydrocholesterol, a sterol present in elevated levels in Smith-Lemli Opitz syndrome, were both significantly more strongly domain-promoting than cholesterol. Domain formation was also affected by the structure of the sphingolipid (or that of an equivalent "saturated" phospholipid) component. Sterols had pronounced effects on domain formation by sphingomyelin and dipalmitoylphosphatidylcholine but only a weak influence on the ability of cerebrosides to form domains. Strikingly it was found that a small amount of ceramide (3 mol %) significantly stabilized domain/raft formation. The molecular basis for, and the implications of, the effects of different sterols and sphingolipids (especially ceramide) on the behavior and biological function of rafts are discussed. PMID- 11432871 TI - N-terminal residues of plasmatocyte-spreading peptide possess specific determinants required for biological activity. AB - Plasmatocyte-spreading peptide (PSP) is a 23-amino acid cytokine that activates a class of insect immune cells called plasmatocytes. The tertiary structure of PSP consists of an unstructured N terminus (residues 1-6) and a well structured core (residues 7-23). A prior study indicated that deletion of the N terminus from PSP eliminated all biological activity. Alanine substitution of the first three residues (Glu(1)-Asn(2)-Phe(3)) further indicated that only replacement of Phe(3) resulted in a loss of activity equal to the N-terminal deletion mutant. Here, we characterized structural determinants of the N terminus. Adding a hydroxyl group to the aromatic ring of Phe(3) (making a Tyr) greatly reduced activity, whereas the addition of a fluorine (p-fluoro) did not. Substitutions that changed the chirality or replaced the aromatic ring of Phe(3) with a branched aliphatic chain (making a Val) also greatly decreased activity. The addition of a methylene group to Val (making a Leu) partially restored activity, whereas the removal of a methylene group from Phe (phenyl-Gly) eliminated all activity. These results indicated that a branched carbon chain with a methylene spacer at the third residue is the minimal structural motif required for activity. The deletion of Glu(1) also eliminated activity. Additional experiments identified the charged N terminal amine and backbone of Glu(1) as key determinants for activity. PMID- 11432872 TI - JAK/STAT3-dependent activation of the RalGDS/Ral pathway in M1 mouse myeloid leukemia cells. AB - The Ras-related GTPase (Ral) is converted to the GTP-bound form by Ral guanine nucleotide dissociation stimulator (RalGDS), a putative effector protein of Ras. Recently, it was proven that Ral regulates c-Src activity and subsequent phosphorylation of its substrate, STAT3. Here, we show that STAT3 inversely regulates activation of Ral through induction of expression of RalGDS. To identify new leukemia inhibitory factor-induced genes, we have performed representational difference analysis using M1 mouse myeloid leukemia cells and cloned RalGDS. The expression of RalGDS and subsequent activation of RalA were clearly suppressed by a dominant negative form of STAT3 and a JAK inhibitor, JAB/SOCS1/SSI-1, indicating that RalGDS/RalA signaling requires the activation of the JAK/STAT3 pathway. An experiment using a Ras inhibitor demonstrated that full activation of RalA also requires activation of Ras. These results suggest a novel cross-talk between JAK/STAT3 and the Ras/RalGDS/Ral signaling pathways through gp130. PMID- 11432873 TI - Schwannomin isoform-1 interacts with syntenin via PDZ domains. AB - The neurofibromatosis type 2 gene (NF2) is involved in the pathogenesis of benign tumors of the human nervous system. The NF2 protein, called schwannomin or merlin, is inactivated in virtually all schwannomas and meningiomas. The molecular mechanisms by which schwannomin functions as a tumor suppressor is unknown but believed to involve plasma membrane-cytoskeletal interactions. Two major alternatively spliced isoforms of schwannomin differing in their C termini have been reported. Using the yeast two-hybrid system, we have identified syntenin as a binding partner for schwannomin isoform-1 (sch-1). Syntenin is an adapter protein that couples transmembrane proteoglycans to cytoskeletal components and is involved in intracellular vesicle transport. The C terminus 25 amino acids of sch-1 and the two PDZ domains of syntenin mediate their binding, and mutations introduced within the VAFFEEL region of sch-1 defined a sequence crucial for syntenin recognition. We have showed that the two proteins interacted in vitro and in vivo and localized underneath the plasma membrane. Fibroblast cells expressing heterologous antisense syntenin display alterations in the subcellular distribution of sch-1. Together, these results provide the first functional clue to the existence of schwannomin isoforms and could unravel novel pathways for the transport and subcellular localization of schwannomin in vivo. PMID- 11432874 TI - Colocalization and interaction of cyclooxygenase-2 with caveolin-1 in human fibroblasts. AB - Results from our previous study suggest that cyclooxygenase-2 (COX-2) induced by phorbol 12-myristate 13-acetate (PMA) may be localized to caveolae-like structures (Liou, J.-Y., Shyue, S.-K., Tsai, M.-J., Chung, C.-L., Chu, K.-Y., and Wu, K. K. (2000) J. Biol. Chem. 275, 15314-15320). In this study, we determined subcellular localization of COX-2 and caveolin-1 by confocal microscopy. COX-2 in human foreskin fibroblasts stimulated by PMA (100 nm) or interleukin-1beta (1 ng/ml) for 6 h was localized to plasma membrane in addition to endoplasmic reticulum and nuclear envelope. Caveolin-1 was localized to plasma membrane, and image overlay showed colocalization of COX-2 with caveolin-1. This was confirmed by the presence of COX-2 and caveolin-1 in the detergent-insoluble membrane fraction of cells stimulated by PMA. Immunoprecipitation showed complex formation of COX-2 with caveolin-1 in a time-dependent manner. A larger quantity of COX-2 was complexed with caveolin-1 in PMA-treated than in interleukin-1beta-treated cells. Purified COX-2 complexed with glutathione S-transferase-fused caveolin-1, which was not inhibited by the scaffolding domain peptide. Caveolin-1-bound COX-2 was catalytically active, and its activity was not inhibited by the scaffolding domain peptide. These results suggest that COX-2 induced by PMA and interleukin 1beta is colocalized with caveolin-1 in the segregated caveolae compartment. Because caveolae are rich in signaling molecules, this COX-2 compartment may play an important role in diverse pathophysiological processes. PMID- 11432875 TI - Membrane targeting by C1 and C2 domains. PMID- 11432876 TI - How membranes shape protein structure. PMID- 11432877 TI - Structure and assembly of beta-barrel membrane proteins. PMID- 11432878 TI - Detergents as tools in membrane biochemistry. AB - Detergents are invaluable tools for studying membrane proteins. However, these deceptively simple, amphipathic molecules exhibit complex behavior when they self associate and interact with other molecules. The phase behavior and assembled structures of detergents are markedly influenced not only by their unique chemical and physical properties but also by concentration, ionic conditions, and the presence of other lipids and proteins. In this minireview, we discuss the various aggregate forms detergents assume and some misconceptions about their structure. The distinction between detergents and the membrane lipids that they may (or may not) replace is emphasized in the most recent high resolution structures of membrane proteins. Detergents are clearly friends and foes, but with the knowledge of how they work, we can use the increasing variety of detergents to our advantage. PMID- 11432879 TI - Membrane protein structural biology minireview series. PMID- 11432880 TI - Copper induces the assembly of a multiprotein aggregate implicated in the release of fibroblast growth factor 1 in response to stress. AB - Fibroblast growth factor (FGF) 1 is known to be released in response to stress conditions as a component of a multiprotein aggregate containing the p40 extravescicular domain of p65 synaptotagmin (Syt) 1 and S100A13. Since FGF1 is a Cu2+-binding protein and Cu2+ is known to induce its dimerization, we evaluated the capacity of recombinant FGF1, p40 Syt1, and S100A13 to interact in a cell free system and the role of Cu2+ in this interaction. We report that FGF1, p40 Syt1, and S100A13 are able to bind Cu2+ with similar affinity and to interact in the presence of Cu2+ to form a multiprotein aggregate which is resistant to low concentrations of SDS and sensitive to reducing conditions and ultracentrifugation. The formation of this aggregate in the presence of Cu2+ is dependent on the presence of S100A13 and is mediated by cysteine-independent interactions between S100A13 and either FGF1 or p40 Syt1. Interestingly, S100A13 is also able to interact in the presence of Cu2+ with Cys-free FGF1 and this observation may account for the ability of S100A13 to export Cys-free FGF1 in response to stress. Lastly, tetrathiomolybdate, a Cu2+ chelator, significantly represses in a dose-dependent manner the heat shock-induced release of FGF1 and S100A13. These data suggest that S100A13 may be involved in the assembly of the multiprotein aggregate required for the release of FGF1 and that Cu2+ oxidation may be an essential post-translational intracellular modifier of this process. PMID- 11432881 TI - Late mortality in childhood cancer: two excellent studies bring good news tempered by room for improvement. PMID- 11432882 TI - Late mortality experience in five-year survivors of childhood and adolescent cancer: the Childhood Cancer Survivor Study. AB - PURPOSE: Survivors of childhood and adolescent cancer are at risk for long-term effects of disease and treatment. The Childhood Cancer Survivor Study assessed overall and cause-specific mortality in a retrospective cohort of 20,227 5-year survivors. PATIENTS AND METHODS: Eligible subjects were individuals diagnosed with cancer (from 1970 to 1986) before the age of 21 who had survived 5 years from diagnosis. Underlying cause of death was obtained from death certificates and other sources and coded and categorized as recurrent disease, sequelae of cancer treatment, or non-cancer-related. Age and sex standardized mortality ratios (SMRs) were calculated using United States population mortality data. RESULTS: The cohort, including 208,947 person-years of follow-up, demonstrated a 10.8-fold excess in overall mortality (95% confidence interval, 10.3 to 11.3). Risk of death was statistically significantly higher in females (SMR = 18.2), individuals diagnosed with cancer before the age of 5 years (SMR = 14.0), and those with an initial diagnosis of leukemia (SMR = 15.5) or CNS tumor (SMR = 15.7). Recurrence of the original cancer was the leading cause of death among 5 year survivors, accounting for 67% of deaths. Statistically significant excess mortality rates were seen due to subsequent malignancies (SMR = 19.4), along with cardiac (SMR = 8.2), pulmonary (SMR = 9.2), and other causes (SMR = 3.3). Treatment-related associations were present for subsequent cancer mortality (radiation, alkylating agents, epipodophyllotoxins), cardiac mortality (chest irradiation, bleomycin), and other deaths (radiation, anthracyclines). No excess mortality was observed for external causes (SMR = 0.8). CONCLUSION: While recurrent disease remains a major contributor to late mortality in 5-year survivors of childhood cancer, significant excesses in mortality risk associated with treatment-related complications exist up to 25 years after the initial cancer diagnosis. PMID- 11432883 TI - Decreasing late mortality among five-year survivors of cancer in childhood and adolescence: a population-based study in the Nordic countries. AB - PURPOSE: To assess the risk of death in patients who survive more than 5 years after diagnosis of childhood cancer and to evaluate causes of death in fatal cases. PATIENTS AND METHODS: This was a population-based study in the five Nordic countries (Denmark, Finland, Iceland, Norway, and Sweden) using data of the nationwide cancer registries and the cause-of-death registries. The study cohort included 13,711 patients who were diagnosed with cancer before the age of 20 years between 1960 and 1989 and who survived at least 5 years from diagnosis. By December 31, 1995, 1,422 patients had died, and death certificates were assessed in 1,402. Standardized mortality ratios (SMRs) for validated causes of death were calculated based on 156,046 patient-years at risk. RESULTS: The overall SMR was 10.8 (95% confidence interval [CI], 10.3 to 11.5), mainly due to high excess mortality from the primary cancer. SMR for second cancer was 4.9 (95% CI, 3.9 to 5.9) and was 3.1 (95% CI, 2.8 to 3.5) for noncancer death. The pattern of causes of death varied markedly between different groups of primary cancer diagnoses and was highly dependent on time passed since diagnosis. Overall late mortality was significantly lower in patients treated during the most recent period of time, 1980 to 1989, compared with those treated from 1960 to 1979 (hazard ratio, 0.61; 95% CI, 0.54 to 0.70), and there was no increase in rates of death due to cancer treatment. CONCLUSION: Long-term survivors of childhood cancer had an increased mortality rate, mainly dying from primary cancers. However, modern treatments have reduced late cancer mortality without increasing the rate of therapy-related deaths. PMID- 11432884 TI - Delay of the diagnostic lumbar puncture and intrathecal chemotherapy in children with acute lymphoblastic leukemia who undergo routine corticosteroid testing: Tokyo Children's Cancer Study Group study L89-12. AB - PURPOSE: To determine the effects of eliminating initial lumbar punctures in 418 consecutively treated children with acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS: Patients were enrolled onto a trial conducted in central Japan between 1989 and 1992. Treatment consisted of standard four-drug induction therapy followed by a risk-based intensification phase, reinduction therapy, late intensification, and remission maintenance therapy (total of 104 weeks). The initial lumbar puncture, with an intrathecal injection of chemotherapy, was performed after 1 week of prednisolone sensitivity testing (day 8). End points included response to prednisolone, CNS status at the time of the day 8 lumbar puncture, subsequent adverse events in CNS and bone marrow, and event-free survival (EFS). RESULTS: The remission induction rate was 93.1% with a 6-year EFS rate (+/- SE) of 68.7% +/- 2.4%, which is similar to historical results for patients who received their diagnostic lumbar puncture and first instillation of intrathecal chemotherapy on day 0. Overall, 84.5% of the patients had good responses to prednisolone, whereas 15.5% had poor responses. Clinical outcome was strikingly better for the good responders (6-year EFS, 74.1% +/- 2.5% compared with 40.1% +/- 6.4% for patients with poor responses), suggesting that omission of intrathecal chemotherapy did not alter the predictive value of drug sensitivity testing. Eighteen patients experienced CNS relapse as their first adverse event (cumulative risk, 5.1%; 95% confidence interval, 2.7% to 7.4%), coincident with reports from groups using conventional strategies of CNS clinical management. Bleeding into the CSF at the time of the day 8 lumbar puncture was apparent in 29 cases (8.1%), but leukemic blasts were identified in only two. CONCLUSION: Delay of the initial lumbar puncture and intrathecal injection of chemotherapy seems to be feasible in children with ALL. Further controlled evaluations are needed to establish the validity of this conclusion. PMID- 11432885 TI - Relapse of TEL-AML1--positive acute lymphoblastic leukemia in childhood: a matched-pair analysis. AB - PURPOSE: The aim of this study was to investigate whether, in relapsed childhood acute lymphoblastic leukemia (ALL), the frequent genetic feature of TEL-AML1 fusion resulting from the cryptic chromosomal translocation t(12;21)(p13;q22) is an independent risk factor. PATIENTS AND METHODS: A matched-pair analysis was performed within a homogeneous group of children with first relapse of BCR-ABL negative B-cell precursor (BPC) ALL treated according to relapse trials ALL Rezidiv (REZ) of the Berlin-Frankfurt-Munster Study Group. A total of 249 patients were eligible for this study: 53 (21%) were positive for TEL-AML1, and 196 (79%) were negative. Positive patients were matched for established most significant prognostic determinants at relapse, time point, and site of relapse, as well as age and peripheral blast cell count at relapse. RESULTS: Fifty pairs matching the aforementioned criteria could be determined. The probabilities with SE of event-free survival and survival at 5 years for matched TEL-AML1 positives and negatives are 0.63 +/- 0.10 versus 0.38 +/- 0.10 (P =.09) and 0.82 +/- 0.09 versus 0.42 +/- 0.19 (P =.10), respectively. These results were confirmed by multivariate analysis, revealing an independent prognostic significance of time point and site of relapse (both P <.001) but not of TEL-AML1 expression (P =.09). CONCLUSION: TEL-AML1 expression does not constitute an independent risk factor in relapsed childhood BCP-ALL after matching for relevant prognostic parameters. It undoubtedly characterizes genetically an ALL entity associated with established favorable prognostic parameters. High-risk therapeutic procedures such as allogeneic SCT should be considered restrictively. PMID- 11432886 TI - Outpatient biochemotherapy with interleukin-2 and interferon alfa-2b in patients with metastatic malignant melanoma: results of two phase II cytokine working group trials. AB - PURPOSE: The Cytokine Working Group performed a randomized phase II trial of two outpatient biochemotherapy regimens to identify an outpatient regimen with high antitumor activity and less toxicity than inpatient regimens which might be compared with chemotherapy or inpatient biochemotherapy regimens in future phase III trials. PATIENTS AND METHODS: Eighty-one patients with metastatic malignant melanoma received dacarbazine 250 mg/m(2)/d intravenously (IV) and cisplatin 25 mg/m(2)/d IV on days 1, 2, and 3, plus interferon (IFN) alfa-2b 5 mU/m(2) subcutaneously (SC) on days 6, 8, 10, 13, and 15, given every 28 days. Interleukin-2 (IL-2) was given daily on days 6 to 10 and 13 to 15. In group 1, IV IL-2 was given at 18.0 MU/m(2), and in group 2, SC IL-2 was given at 5.0 mU/m(2). RESULTS: In group 1 (IV IL-2), there were five complete responses (CRs) and 11 partial responses (PRs) among 44 patients (objective response rate [ORR], 36%; 95% confidence interval [CI], 22% to 51%). In group 2 (SC IL-2), there was one CR and five PRs among the 36 patients (ORR, 17%; 95% CI, 4% to 29%). The median survival was 10.7 months in group 1 and 7.3 months in group 2. Eleven patients in group 1 and four patients in group 2 remain alive as of the last follow-up. Toxicities in both groups were similar. No patient required hospitalization for neutropenic fever. CONCLUSION: Biochemotherapy has activity in these outpatient regimens with acceptable toxicity. The antitumor activity observed with the IV IL 2 regimen seems similar to that of inpatient biochemotherapy regimens. If inpatient biochemotherapy regimens develop an established role in the management of melanoma, future phase III trial comparisons with this outpatient IV IL-2 regimen would be appropriate. PMID- 11432887 TI - Treatment-induced pathologic necrosis: a predictor of local recurrence and survival in patients receiving neoadjuvant therapy for high-grade extremity soft tissue sarcomas. AB - PURPOSE: To determine whether treatment-induced pathologic necrosis correlates with local recurrence and overall survival in patients who receive neoadjuvant therapy for high-grade extremity soft tissue sarcomas. PATIENTS AND METHODS: Four hundred ninety-six patients with intermediate- to high-grade extremity soft tissue sarcomas received protocol neoadjuvant therapy. All patients underwent surgical resection after neoadjuvant therapy and had pathologic assessment of tumor necrosis in the resected specimens. RESULTS: The 5- and 10-year local recurrence rates for patients with > or = 95% pathologic necrosis were significantly lower (6% and 11%, respectively) than the local recurrence rates for patients with less than 95% pathologic necrosis (17% and 23%, respectively). The 5- and 10-year survival rates for the patients with > or = 95% pathologic necrosis were significantly higher (80% and 71%, respectively) than the survival rates for the patients with less than 95% pathologic necrosis (62% and 55%, respectively). Patients with less than 95% pathologic necrosis were 2.51 times more likely to develop a local recurrence and 1.86 times more likely to die of their disease as compared with patients with > or = 95% pathologic necrosis. The percentage of patients who achieved > or /= 95% pathologic necrosis increased to 48% with the addition of ifosfamide as compared with 13% of the patients in all the other protocols combined. CONCLUSION: Treatment-induced pathologic necrosis is an independent predictor of both local recurrence and overall survival in patients who receive neoadjuvant therapy for high-grade extremity soft tissue sarcomas. A complete pathologic response (> or = 95% pathologic necrosis) correlated with a significantly lower rate of local recurrence and improved overall survival. PMID- 11432888 TI - Randomized phase III trial of paclitaxel plus carboplatin versus vinorelbine plus cisplatin in the treatment of patients with advanced non--small-cell lung cancer: a Southwest Oncology Group trial. AB - PURPOSE: This randomized trial was designed to determine whether paclitaxel plus carboplatin (PC) offered a survival advantage over vinorelbine plus cisplatin (VC) for patients with advanced non--small-cell lung cancer. Secondary objectives were to compare toxicity, tolerability, quality of life (QOL), and resource utilization. PATIENTS AND METHODS: Two hundred two patients received VC (vinorelbine 25 mg/m(2)/wk and cisplatin 100 mg/m(2)/d, day 1 every 28 days) and 206 patients received PC (paclitaxel 225 mg/m(2) over 3 hours with carboplatin area under the curve of 6, day 1 every 21 days). Patients completed QOL questionnaires at baseline, 13 weeks, and 25 weeks. Resource utilization forms were completed at five time points through 24 months. RESULTS: Patient characteristics were similar between the groups. The objective response rate was 28% in the VC arm and 25% in the PC arm. Median survival was 8 months in both arms, with 1-year survival rates of 36% and 38%, respectively. Grade 3 and 4 leukopenia (P =.002) and neutropenia (P =.008) occurred more frequently on the VC arm. Grade 3 nausea and vomiting were higher on the VC arm (P =.001, P =.007), and grade 3 peripheral neuropathy was higher on the PC arm (P <.001). More patients on the VC arm discontinued therapy because of toxicity (P =.001). No difference in QOL was observed. Overall costs on the PC arm were higher than on the VC arm because of drug costs. CONCLUSION: PC is equally efficacious as VC for the treatment of advanced non--small-cell lung cancer. PC is less toxic and better tolerated but more expensive than VC. New treatment strategies should be pursued. PMID- 11432889 TI - Estimating the cost of informal caregiving for elderly patients with cancer. AB - PURPOSE: As the United States population ages, the increasing prevalence of cancer is likely to result in higher direct medical and nonmedical costs. Although estimates of the associated direct medical costs exist, very little information is available regarding the prevalence, time, and cost associated with informal caregiving for elderly cancer patients. MATERIALS AND METHODS: To estimate these costs, we used data from the first wave (1993) of the Asset and Health Dynamics (AHEAD) Study, a nationally representative longitudinal survey of people aged 70 or older. Using a multivariable, two-part regression model to control for differences in health and functional status, social support, and sociodemographics, we estimated the probability of receiving informal care, the average weekly number of caregiving hours, and the average annual caregiving cost per case (assuming an average hourly wage of $8.17) for subjects who reported no history of cancer (NC), having a diagnosis of cancer but not receiving treatment for their cancer in the last year (CNT), and having a diagnosis of cancer and receiving treatment in the last year (CT). RESULTS: Of the 7,443 subjects surveyed, 6,422 (86%) reported NC, 718 (10%) reported CNT, and 303 (4%) reported CT. Whereas the adjusted probability of informal caregiving for those respondents reporting NC and CNT was 26%, it was 34% for those reporting CT (P <.05). Those subjects reporting CT received an average of 10.0 hours of informal caregiving per week, as compared with 6.9 and 6.8 hours for those who reported NC and CNT, respectively (P <.05). Accordingly, cancer treatment was associated with an incremental increase of 3.1 hours per week, which translates into an additional average yearly cost of $1,200 per patient and just over $1 billion nationally. CONCLUSION: Informal caregiving costs are substantial and should be considered when estimating the cost of cancer treatment in the elderly. PMID- 11432890 TI - Late medical complications and fatigue in Hodgkin's disease survivors. AB - PURPOSE: Long-term medical complications, such as cardiac, pulmonary, and thyroid dysfunction, are frequent among Hodgkin's disease survivors (HDSs). Chronic fatigue is also highly prevalent among HDSs. Few studies have explored possible etiologic explanations for fatigue. The aim of this study was to explore whether late cardiac, pulmonary, and thyroid complications after curative treatment for Hodgkin's disease (HD) may explain the high level of fatigue among HDSs. PATIENTS AND METHODS: Four-hundred fifty-nine patients treated for HD at the Norwegian Radium Hospital from 1971 to 1991 were included in a cross-sectional, follow-up study of subjective health status. Fatigue (physical [PF] and mental), was measured by the Fatigue Questionnaire. A subcohort of the HDSs (116 patients) treated from 1980 to 1988 were included in a separate study in which long-term cardiac, pulmonary, and thyroid complications were assessed. All patients had received radiotherapy, and 63 patients had received additional chemotherapy. The present study comprised 92 patients (mean age, 37 years; range, 23 to 56 years) who participated in both studies. RESULTS: HDSs with pulmonary dysfunction were more fatigued than HDSs with normal pulmonary function (PF 10.9 v 8.9; P <.05). Gas transfer impairment was the most prevalent pulmonary dysfunction, and three times as many patients with gas transfer impairment reported chronic fatigue (duration, 6 months or longer), compared with patients without pulmonary dysfunction (48% v 17%, P <.01). No associations were found between cardiac sequelae or hypothyroidism and fatigue. CONCLUSION: Pulmonary dysfunction is associated with fatigue in HDSs. Cardiac sequelae was not associated with fatigue in HDSs. We question the absence of an association between thyroid complications and fatigue. PMID- 11432891 TI - Phase I study of anti--epidermal growth factor receptor antibody cetuximab in combination with radiation therapy in patients with advanced head and neck cancer. AB - PURPOSE: To evaluate the safety, pharmacokinetics, and efficacy of a chimeric anti-epidermal growth factor receptor monoclonal antibody, cetuximab, in combination with radiation therapy (RT) in patients with advanced squamous cell carcinoma of the head and neck. PATIENTS AND METHODS: We treated 16 patients in five successive treatment schedules. A standard dose escalation procedure was used; three patients entered onto the study at each dose level of cetuximab received conventional RT (70 Gy, 2 Gy/d), and the final three patients received hyperfractionated RT (76.8 Gy, 1.2 Gy bid). Cetuximab was delivered as a loading dose of 100 to 500 mg/m(2), followed by weekly infusions of 100 to 250 mg/m(2) for 7 to 8 weeks. Circulating levels of cetuximab during therapy were determined using a biomolecular interaction analysis core instrument. Human antichimeric antibody response was evaluated with a double-antigen radiometric assay. The recommended phase II/III dose was defined as the optimal cetuximab dose level based on the pharmacologic parameters and adverse events. RESULTS: The most commonly reported adverse events were fever, asthenia, transaminase elevation, nausea, and skin toxicities (grade 1 to 2 in most patients). Skin toxicity outside of the RT field was not strictly dose-dependent; however, grade 2 or higher events were observed in patients treated with higher dose regimens. There was one grade 4 allergic reaction. Most acute adverse effects were associated with RT (xerostomia, mucositis, and local skin toxicity). No antibodies against cetuximab were detected. All patients achieved an objective response (13 complete and two partial remissions). CONCLUSION: Cetuximab can be safely administered with RT. The recommended dose for phase II/III studies is a loading dose of 400 to 500 mg/m(2) and a maintenance weekly dose of 250 mg/m(2). PMID- 11432892 TI - Efficacy and safety of gemtuzumab ozogamicin in patients with CD33-positive acute myeloid leukemia in first relapse. AB - PURPOSE: Three open-label, multicenter trials were conducted to evaluate the efficacy and safety of single-agent Mylotarg (gemtuzumab ozogamicin; CMA-676; Wyeth Laboratories, Philadelphia, PA), an antibody-targeted chemotherapy agent, in patients with CD33-positive acute myeloid leukemia (AML) in untreated first relapse. PATIENTS AND METHODS: The study population comprised 142 patients with AML in first relapse with no history of an antecedent hematologic disorder and a median age of 61 years. All patients received Mylotarg as a 2-hour intravenous infusion, at a dose of 9 mg/m(2), at 2-week intervals for two doses. Patients were evaluated for remission, survival, and treatment-emergent adverse events. RESULTS: Thirty percent of patients treated with Mylotarg obtained remission as characterized by 5% or less blasts in the marrow, recovery of neutrophils to at least 1,500/microL, and RBC and platelet transfusion independence. Although patients treated with Mylotarg had relatively high incidences of myelosuppression, grade 3 or 4 hyperbilirubinemia (23%), and elevated hepatic transaminase levels (17%), the incidences of grade 3 or 4 mucositis (4%) and infections (28%) were relatively low. There was a low incidence of severe nausea and vomiting (11%) and no treatment-related cardiotoxicity, cerebellar toxicity, or alopecia. Many patients received Mylotarg on an outpatient basis (38% and 41% of patients for the first and second doses, respectively). Among the 142 patients, the median total duration of hospitalization was 24 days; 16% of patients required 7 days of hospitalization or less. CONCLUSION: Administration of the antibody-targeted chemotherapy agent Mylotarg to patients with CD33 positive AML in first relapse induces complete remissions with what appears to be a favorable safety profile. PMID- 11432893 TI - Effective dosing of topotecan with carboplatin in relapsed ovarian cancer: a phase I/II study. AB - PURPOSE: This phase I/II study was performed to evaluate the feasibility of administering the topoisomerase inhibitor topotecan in combination with carboplatin. PATIENTS AND METHODS: Topotecan was given as a 30-minute infusion daily for 5 days, with carboplatin given immediately after topotecan on day 5. Treatment was repeated every 21 days. Carboplatin and then topotecan were escalated in sequential cohorts of three to six patients. Four dosage combinations of topotecan days 1 to 5 and carboplatin (day 5) were tested: 0.5 mg/m(2)/d and carboplatin area under the curve (AUC) of 4, topotecan 0.5 mg/m(2)/d and carboplatin AUC of 5, topotecan 0.75 mg/m(2)/d and carboplatin AUC of 5, and topotecan 1.0 mg/m(2)/d and carboplatin AUC of 5. RESULTS: Grade 3 and 4 neutropenia was common at doses of 0.75 mg/m(2)/d and above, but dose-limiting hematologic toxicity occurred in only one patient. The most common reason for dose reduction or delay was failure of myelosuppression to resolve by day 21. Nonhematologic toxicity was generally mild. The maximum-tolerated dose as defined in the protocol was not reached, but topotecan dose escalation was stopped at 1.0 mg/m(2)/d, because delayed neutrophil recovery precluded re-treatment on a 21-day schedule. CONCLUSION: Hematologic toxicity was common but rarely serious, and the combination of topotecan with carboplatin on this schedule was safe and well tolerated. Giving carboplatin to patients after topotecan on day 5, rather than on day 1, allowed dose escalation beyond the levels reported in other studies. The recommended doses for previously treated patients are topotecan 0.75 mg/m(2)/d, days 1 to 5, with carboplatin at an area under the curve (AUC) of 5 following topotecan on day 5. The combination of topotecan 1 mg/m(2)/d, days 1 to 5, followed on day 5 by carboplatin at an AUC of 5, merits further examination in untreated patients. PMID- 11432894 TI - Toxicity, efficacy, and pharmacology of suramin in adults with recurrent high grade gliomas. AB - PURPOSE: To determine the toxicity, efficacy, and pharmacology of suramin in patients with recurrent or progressive recurrent high-grade gliomas. PATIENTS AND METHODS: Fifty adults were to receive suramin. However, if no responses were seen in the first ten patients, the study was to be terminated. A total of 12 patients were enrolled onto this trial. Ten patients had glioblastoma multiforme, and 11 had received prior nitrosoureas. RESULTS: Drug-related toxicities were modest and reversible. Three patients developed grade 3 to 4 neutropenia, constipation, diarrhea, or nausea. No CNS bleeding was observed. Median time to progression was 55 days (range, 17 to 242 days) and median survival was 191 days (range, 42 to 811 days). No partial or complete responses were seen at 12 weeks. However, the clinical outcome of three patients suggests that evidence of suramin activity may be delayed. One patient who "progressed" after 12 weeks of suramin had a subsequent marked reduction in tumor size and has maintained an excellent partial response for over 2 years without other therapy. Two others had disease stabilization and lived for 16 and 27 months. Pharmacokinetics from 11 patients revealed that all reached target suramin concentrations. CONCLUSION: This study demonstrates that suramin is well tolerated by patients with recurrent high-grade gliomas and may have efficacy in this disease. Its pharmacology seems unaffected by anticonvulsants. As a result of this data, suramin and radiation are now being administered concurrently to patients with newly diagnosed glioblastoma multiforme, with survival as the primary outcome. PMID- 11432895 TI - Phase I and pharmacologic study of OSI-774, an epidermal growth factor receptor tyrosine kinase inhibitor, in patients with advanced solid malignancies. AB - PURPOSE: To assess the feasibility of administering OSI-774, to recommend a dose on a protracted, continuous daily schedule, to characterize its pharmacokinetic behavior, and to acquire preliminary evidence of anticancer activity. PATIENTS AND METHODS: Patients with advanced solid malignancies were treated with escalating doses of OSI-774 in three study parts (A to C) to evaluate progressively longer treatment intervals. Part A patients received OSI-774 25 to 100 mg once daily, for 3 days each week, for 3 weeks every 4 weeks. Part B patients received OSI-774 doses ranging from 50 to 200 mg given once daily for 3 weeks every 4 weeks to establish the maximum tolerated dose (MTD). In part C, patients received this MTD on a continuous, uninterrupted schedule. The pharmacokinetics of OSI-774 and its O-demethylated metabolite, OSI-420, were characterized. RESULTS: Forty patients received a total of 123 28-day courses of OSI-774. No severe toxicities precluded dose escalation of OSI-774 from 25 to 100 mg/d in part A. In part B, the incidence of severe diarrhea and/or cutaneous toxicity was unacceptably high at OSI-774 doses exceeding 150 mg/d. Uninterrupted, daily administration of OSI-774 150 mg/d represented the MTD on a protracted daily schedule. The pharmacokinetics of OSI-774 were dose independent; repetitive daily treatment did not result in drug accumulation (at 150 mg/d [average]: minimum steady-state plasma concentration, 1.20 +/- 0.62 microg/mL; clearance rate, 6.33 +/- 6.41 L/h; elimination half-life, 24.4 +/- 14.6 hours; volume of distribution, 136. 4 +/- 93.1 L; area under the plasma concentration time curve for OSI-420 relative to OSI-774, 0.12 +/- 0.12 microg/h/mL). CONCLUSION: The recommended dose for disease-directed studies of OSI-774 administered orally on a daily, continuous, uninterrupted schedule is 150 mg/d. OSI-774 was well tolerated, and several patients with epidermoid malignancies demonstrated either antitumor activity or relatively long periods of stable disease. The precise contribution of OSI-774 to these effects is not known. PMID- 11432896 TI - Phase I study of recombinant human CD40 ligand in cancer patients. AB - PURPOSE: To determine the toxicity, maximum-tolerated dose (MTD), and pharmacokinetics of recombinant human CD40 ligand (rhuCD40L) (Avrend; Immunex Corp, Seattle, WA), suggested in preclinical studies to mediate cytotoxicity against CD40-expressing tumors and immune stimulation. PATIENTS AND METHODS: Patients with advanced solid tumors or intermediate- or high-grade non-Hodgkin's lymphoma (NHL) received rhuCD40L subcutaneously daily for 5 days in a phase I dose-escalation study. Subsequent courses were given until disease progression. RESULTS: Thirty-two patients received rhuCD40L at three dose levels. A total of 65 courses were administered. The MTD was 0.1 mg/kg/d based on dose-related but transient elevations of serum liver transaminases. Grade 3 or 4 transaminase elevations occurred in 14%, 28%, and 57% of patients treated at 0.05, 0.10, and 0.15 mg/kg/d, respectively. Other toxicities were mild to moderate. At the MTD, the half-life of rhuCD40L was calculated at 24.8 +/- 22.8 hours. Two patients (6%) had a partial response on study (one patient with laryngeal carcinoma and one with NHL). For the patient with laryngeal cancer, a partial response was sustained for 12 months before the patient was taken off therapy and observed on no additional therapy. Three months later, the patient was found to have a complete response and remains biopsy-proven free of disease at 24 months. Twelve patients (38%) had stable disease after one course, which was sustained in four patients through four courses. CONCLUSION: The MTD of rhuCD40L when administered subcutaneously daily for 5 days was defined by transient serum elevations in hepatic transaminases. Encouraging antitumor activity, including a long-term complete remission, was observed. Phase II studies are warranted. PMID- 11432897 TI - Unusual presentations of malignancy. Case 1. Male inflammatory (?) breast cancer. PMID- 11432898 TI - Unusual presentations of malignancy. Case 2. Breast plasmacytoma in a patient with human immunodeficiency virus. PMID- 11432899 TI - Unusual presentations of malignancy. Case 3. Mantle-cell lymphoma of soft tissue. PMID- 11432900 TI - "But doctor, what have i got to lose...?". PMID- 11432901 TI - Meningeal carcinomatosis from breast carcinoma responsive to trastuzumab. PMID- 11432902 TI - Early-stage Hodgkin's disease: to mantle or not to mantle? PMID- 11432903 TI - Angiogenesis in cancer. PMID- 11432904 TI - Discussing do-not-resuscitate status: furthering the discourse. PMID- 11432905 TI - A role for 'futile cycles' involving invertase and sucrose synthase in sucrose metabolism of tomato fruit. AB - Current concepts of the factors determining sink strength and the subsequent regulation of carbohydrate metabolism in tomato fruit are based upon an understanding of the relative roles of sucrose synthase, sucrose phosphate synthase and invertase, derived from studies in mutants and transformed plants. These enzymes participate in at least four futile cycles that involve sugar transport between the cytosol, vacuole and apoplast. Key reactions are (1) the continuous rapid degradation of sucrose in the cytosol by sucrose synthase (SuSy), (2) sucrose re-synthesis via either SuSy or sucrose phosphate synthase (SPS), (3) sucrose hydrolysis in the vacuole or apoplast by acid invertase, (4) subsequent transport of hexoses to the cytosol where they are once more converted into sucrose, and (5) rapid synthesis and breakdown of starch in the amyloplast. In this way futile cycles of sucrose/hexose interchange govern fruit sugar content and composition. The major function of the high and constant invertase activity in red tomato fruit is, therefore, to maintain high cellular hexose concentrations, the hydrolysis of sucrose in the vacuole and in the intercellular space allowing more efficient storage of sugar in these compartments. Vacuolar sugar storage may be important in sustaining fruit cell growth at times when less sucrose is available for the sink organs because of exhaustion of the carbohydrate pools in source leaves. PMID- 11432906 TI - The pathways of calcium movement to the xylem. AB - Calcium is an essential plant nutrient. It is acquired from the soil solution by the root system and translocated to the shoot via the xylem. The root must balance the delivery of calcium to the xylem with the need for individual root cells to use [Ca2+]cyt for intracellular signalling. Here the evidence for the current hypothesis, that Ca2+ travels apoplastically across the root to the Casparian band which it then circumvents via the cytoplasm of the endodermal cell, is critically reviewed. It is noted that, although Ca2+ channels and Ca2+ ATPases are present and could catalyse Ca2+ influx and efflux across the plasma membrane of endodermal cells, their transport capacity is unlikely to be sufficient for xylem loading. Furthermore, there seems to be no competition, or interactions, between Ca2+, Ba2+ and Sr2+ for transport to the shoot. This seems incompatible with a symplastic pathway involving at least two protein-catalysed transport steps. Thus, a quantity of purely apoplastic Ca2+ transport to the xylem is indicated. The relative contributions of these two pathways to the delivery of Ca2+ to the xylem are unknown. However, the functional separation of symplastic Ca2+ fluxes (for root nutrition and cell signalling) and apoplastic Ca2+ fluxes (for transfer to the shoot) would enable the root to fulfil the demand of the shoot for calcium without compromising intracellular [Ca2+]cyt signals. This is also compatible with the observed correlation between transpiration rate and calcium delivery to the shoot. PMID- 11432907 TI - In vitro tolerance to Botrytis cinerea of grapevine 41B rootstock in transgenic plants expressing the stilbene synthase Vst1 gene under the control of a pathogen inducible PR 10 promoter. AB - Resveratrol is a major phytoalexin in grapevine but its synthesis in response to phytopathogen attack decreases with grape berry ripening. A chimeric gene combining an alfalfa PR 10 promoter and Vst1 (Vitis stilbene synthase 1) gene was introduced into the genome of 41B rootstock. Transgenic plants were analysed for resveratrol production in leaves infected with Botrytis using an in vitro test. Among the 50 transgenic lines analysed, some exhibited a production lower than the non-transgenic control, but others accumulated resveratrol from 5-100-fold. Moreover, in the latter clones, symptoms were highly reduced in response to infection. These results were a good indication that the combination of a pathogen-inducible promoter and a defence gene may increase tolerance against fungi in grapevine. The efficacy of this approach should be further tested by experiments conducted in the vineyard. PMID- 11432908 TI - Deglycosylation is necessary but not sufficient for activation of proconcanavalin A. AB - Concanavalin A (ConA), one of the most studied plant lectins, is formed in jack bean (Canavalia ensiformis) seeds. ConA is synthesized as an inactive glycoprotein precursor proConA. Different processing events such as endoproteolytic cleavages, ligation of peptides and deglycosylation of the precursor are required to generate the different polypeptides constitutive of mature ConA. Among these events, deglycosylation of the prolectin appears as a key step in the lectin activation. The detection of deglycosylated proConA in immature jack bean seeds indicates that endoproteolytic cleavages are not prerequisite for its deglycosylation. Both the structure of the lectin precursor N-glycans Man8-9GlcNAc2 and the capacity of Endo H to cleave these oligosaccharide from native proConA in vitro favoured Endo H-type glycosidases as candidates for proConA deglycosylation in planta. Evidence for pH-dependent changes in the prolectin folding were obtained from analysis of the N-glycan accessibility and activation of the deglycosylated lectin precursor in acidic conditions. These data are consistent with the observation that both deglycosylation and acidification of the pH are the minimum requirements to convert the inactive precursor into an active lectin. PMID- 11432909 TI - Viability loss of neem (Azadirachta indica) seeds associated with membrane phase behaviour. AB - Storage of neem (Azadirachta indica) seeds is difficult because of their sensitivity to chilling stress at moisture contents (MC) > or =10% or imbibitional stress below 10% MC. The hypothesis was tested that an elevated gel to-liquid crystalline phase transition temperature (Tm) of membranes is responsible for this storage behaviour. To this end a spin probe technique, Fourier transform infrared microspectroscopy, and electron microscopy were used. The in situ Tm of hydrated membranes was between 10 degrees C and 15 degrees C, coinciding with the critical minimum temperature for germination. During storage, viability of fresh embryos was lost within two weeks at 5 degrees C, but remained high at 25 degrees C. The loss of viability coincided with an increased leakage of K+ from the embryos upon imbibition and with an increased proportion of cells with injured plasma membranes. Freeze-fracture replicas of plasma membranes from chilled, hydrated axes showed lateral phase separation and signs of the inverted hexagonal phase. Dehydrated embryos were sensitive to soaking in water, particularly at low temperatures, but fresh embryos were not. After soaking dry embryos at 5 degrees C (4 h) plus 1 d of further incubation at 25 degrees C, the axis cells were structurally disorganized and did not become turgid. In contrast, cells had a healthy appearance and were turgid after soaking at 35 degrees C. Imbibitional stress was associated with the loss of plasma membrane integrity in a limited number of cells, which expanded during further incubation of the embryos at 25 degrees C. It is suggested that the injuries brought about by storage or imbibition at sub-optimal temperatures in tropical seeds whose membranes have a high intrinsic Tm (10-15 degrees C), are caused by gel phase formation. PMID- 11432910 TI - Changes in fatty acid composition during development of tissues of coconut (Cocos nucifera L.) embryos in the intact nut and in vitro. AB - Intact coconuts were germinated in situ and compared with excised zygotic embryos germinated in vitro. The growth of the embryonic tissue and their fatty acid compositions were measured. Haustoria, plumules and radicles of coconuts germinated in situ grew continuously and proportionately throughout the 120 d experiment with haustauria increasing to 45 g x nut(-1) and weighing 4-5-fold more than the other two tissues. The plumules and radicles of the seedlings cultured in vitro also grew continuously but the haustoria grew sporadically between 15 d and 75 d in culture and, at 250 mg x nut(-1) after 75 d, were smaller than the other two tissues. All the tissues of the nuts grown in situ contained significant amounts of lauric acid, the acid characteristic of coconut oil, as well as longer chain saturated and unsaturated fatty acids. The content of medium and long chain fatty acids increased in all growing tissues as the experiment proceeded, especially the haustorium which contained 24-35% of its fatty acid as lauric acid; the fat content of solid endosperm reduced during this period. Seedlings grown in vitro, on the other hand, failed to accumulate lauric acid in any of their tissues (haustorium contained 6-11% of its fatty acid as lauric acid). The results may have implications for the design of growth media for growing zygotic and somatic cultures of coconut and may provide a marker for successful germination. PMID- 11432911 TI - Trehalose becomes the most abundant non-structural carbohydrate during senescence of soybean nodules. AB - Carbohydrate metabolism and symbiont survival were studied in nodules of soybean (G. max [L.] Merr. cv. Maple Arrow infected with Bradyrhizobium japonicum 61-A 101), induced to senesce simultaneously by application of the photosynthesis inhibitor dichloromethyl urea (DCMU). The plant-borne carbohydrates sucrose and starch started to decline after 2 d and reached background levels after 8 d, in parallel with the decline of nitrogenase. However, the microsymbiont-borne disaccharide trehalose declined only by about 40% and subsequently remained at a constant level of c. 6 mg x g(-1) dry weight up to 14 d, when nodules softened and decayed. The number of re-isolated viable bacteria was not significantly decreased in senescent nodules as compared to control nodules. These results indicate that during terminal senescence of nodules an appreciable part of the bacteria conserve their trehalose pools and survive. PMID- 11432912 TI - Characterization of position-induced spatial and temporal regulation of transgene promoter activity in plants. AB - Quantitative differences in transgene expression between independent transformants are generally ascribed to different integration sites of the transgene (position effect). The contribution of spatial and temporal changes in transgene promoter activity to these position-induced differences in transgene expression in planta are characterized, using the firefly luciferase (luc) reporter system. The activity of three different promoters (Cauliflower Mosaic Virus (CaMV) 35S, modified CaMV 35S and the promoter of an Arabidopsis thaliana Lipid Transfer Protein gene) was shown to vary not only among independent transformants, but also between leaves on the same plant and within a leaf. The differences in local LUC activity between leaves and within a leaf correlated with differences in local luc mRNA steady-state levels. Imaging of LUC activity in the same leaves over a 50 d period, shows that individual transformants can show different types of temporal regulation. Both the spatial and the temporal type of luc transgene expression pattern are inherited by the next generation. It is concluded that previously reported position-induced quantitative differences in transgene expression are probably an accumulated effect of differences in spatial and temporal regulation of transgene promoter activity. PMID- 11432913 TI - Changes in leaf hexokinase activity and metabolite levels in response to drying in the desiccation-tolerant species Sporobolus stapfianus and Xerophyta viscosa. AB - The phosphorylation of glucose and fructose is an important step in regulating the supply of hexose sugars for biosynthesis and metabolism. Changes in leaf hexokinase (EC 2.7.1.1) activity and in vivo metabolite levels were examined during drying in desiccation-tolerant Sporobolus stapfianus and Xerophyta viscosa. Leaf hexokinase activity was significantly induced from 85% to 29% relative water content (RWC) in S. stapfianus and from 89% to 55% RWC in X. viscosa. The increase in hexokinase corresponded to the region of sucrose accumulation in both species, with the highest activity levels coinciding with region of net glucose and fructose removal. The decline of hexose sugars and accumulation of sucrose in both plant species was not associated with a decline in acid and neutral invertase. The increase in hexokinase activity may be important to ensure that the phosphorylation and incorporation of glucose and fructose into metabolism exceeded production from potential hydrolytic activity. Total cellular glucose-6-phosphate (Glc-6-P) and fructose-6-phosphate (Fru-6-P) levels were held constant throughout dehydration. In contrast to hexokinase, fructokinase activity was unchanged during dehydration. Hexokinase activity was not fully induced in leaves of S. stapfianus dried detached from the plant, suggesting that the increase in hexokinase may be associated with the acquisition of desiccation-tolerance. PMID- 11432914 TI - Enhanced tolerance of transgenic poplar plants overexpressing gamma glutamylcysteine synthetase towards chloroacetanilide herbicides. AB - A wild-type poplar hybrid and two transgenic clones overexpressing a bacterial gamma-glutamylcysteine synthetase in the cytosol or in the chloroplasts were exposed to the chloroacetanilide herbicides acetochlor and metolachlor dispersed in the soil. The transformed poplars contained higher gamma-glutamylcysteine and glutathione (GSH) levels than wild-type plants and therefore it was supposed that they would have an elevated tolerance towards these herbicides, which are detoxified in GSH-dependent reactions. Phenotypically, the transgenic and wild type plants did not differ. The growth and the biomass of all poplar lines were markedly reduced by the two chloroacetanilide herbicides. However, the decrease of shoot and root fresh weights caused by the herbicides was significantly smaller in the transgenic than in wild-type plants. In addition, the growth rate of poplars transformed in the cytosol was reduced to a significantly lesser extent than that of wild-type plants following herbicide treatments. The effects of the two herbicides were similar. Herbicide exposures markedly increased the levels of gamma-glutamylcysteine and GSH in leaves of each poplar line. The increase in the foliar amounts of these thiols was stronger in the transgenic lines than in the wild type, particularly in the upper leaves. Considerable GST activities were detected in leaves of all poplar plants. Exposure of poplars to chloroacetanilide herbicides resulted in a marked induction of GST activity in upper leaf positions but not in middle and lower leaves. The extent of enzyme induction did not differ significantly between transgenic and wild-type poplars. Although the results show that the transgenic poplar lines are good candidates for phytoremediation purposes, the further improvement of their detoxification capacity, preferably by transformation using genes encoding herbicide-specific GST isoenzymes, seems to be the most promising way to obtain plants suitable for practical application. PMID- 11432915 TI - Induction of air embolism in xylem conduits of pre-defined diameter. AB - A new method is presented that enables the induction of embolisms in a fraction of all xylem vessels, based on diameter, at one cut end of a stem segment. The method is based on the different capillary characteristic of xylem vessels of different cross-sectional size. To verify the method, air embolisms were induced in cut xylem vessels of chrysanthemum (Dendranthemaxgrandiflorum Tzvelev cv. Cassa) stem segments at different xylem tensions and compared with the distribution of gas-filled vessels as visualized by cryo-scanning electron microscopy (Cryo-SEM). At -6 kPa xylem pressure, air-entrance was only induced in large diameter vessels (>30 microm), while at -24 kPa embolisms were induced in almost all xylem vessels (>10 microm). Although the principle of the embolization method worked well, smaller diameter vessels were observed to be embolized than was expected according to the calculations. The role of cross-sectional shape and contact angle between xylem sap and vessel wall at the menisci are discussed. After correction for the observed (diameter independent) deviation from circularity of the cross-sectional vessel shape the contact angle was calculated to be approximately 55 degrees. Hydraulic resistance (Rh) measurements before and after embolization showed that the effect of embolizing only large diameter cut xylem vessels had only a small influence on overall Rh of a stem segment. Embolizing all cut xylem vessels at one cut end almost trebled overall Rh. The difference was discussed in the light of the networking capacity of the xylem system. PMID- 11432916 TI - Leaf habit influences nitrogen remobilization in Vaccinium species. AB - The effect of N supply on plant growth and leaf demography of a deciduous and an evergreen Ericaceae was studied in relation to their internal cycling of N. Mature ramets of Vaccinium myrtillus (deciduous) and Vaccinium vitis-idaea (evergreen) were established in sand culture for 1 year with an adequate supply of a balanced nutrient solution. During one growing season, the plants were given two levels of N supply enriched with 15N and eight sequential destructive harvests were taken. Recovery of unlabelled N in the new shoot was used to determine the remobilization of N from storage. Initially, growth was unaffected by N supply. After May, High N enhanced growth for both species but the nature of their growth response differed. For both species, new shoot biomass and leaf number increased but root biomass production was affected for V. myrtillus only. Whole plant biomass production was similar for both species under High N, but was greater for V. vitis-idaea under Low N. The amount of N remobilized to support new shoot growth was similar for the two species and was independent of N current supply. N was remobilized predominantly from previous year leaves for V. vitis idaea and from previous year stems and roots for V. myrtillus. The contribution of remobilization to new shoot N was similar for the two species, but depended on N supply. Remobilization was faster in V. myrtillus, but lasted longer in V. vitis-idaea. The results are discussed in relation to species growth in N-poor environments, focusing on the extent to which species-differences in the dynamic of N remobilization and growth may explain their adaptation to constant and/or changeable N supply. PMID- 11432917 TI - Ultrastructural localization of photosynthetic and photorespiratory enzymes in epidermal, mesophyll, bundle sheath, and vascular bundle cells of the C4 dicot Amaranthus viridis. AB - In the leaves of the NAD-malic enzyme (NAD-ME)-type C4 dicot Amaranthus viridis L., there are chloroplasts in the vascular parenchyma cells (VPC), companion cells (CC), ordinary epidermal cells (EC), and guard cells (GC), as well as in the mesophyll cells (MC) and the bundle sheath cells (BSC). However, the chloroplasts of the VPC, CC, EC, and GC are smaller than those of the MC and BSC. In this study, the accumulation of photosynthetic and photorespiratory enzymes in these leaf cell types was investigated by immunogold labelling and electron microscopy. Strong labelling for phosphoenolpyruvate carboxylase was found in the MC cytosol. Weak labelling was observed in the CC and GC cytosol. Labelling for pyruvate, Pi dikinase occurred to varying degrees in the chloroplasts of all cell types except CC. Labelling for the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase was detected in the chloroplasts of all cell types except MC. For both NAD-ME and the P-protein of glycine decarboxylase, intense labelling was found in the BSC mitochondria; weaker labelling was recognized in the VPC mitochondria. These data indicate that when not only the MC and BSC but also the other leaf cell types are included, the cell-specific expression of the enzymes in C4 leaves becomes more complex than has been known previously. These findings are discussed in relation to the metabolic function of epidermal and vascular bundle cells. PMID- 11432918 TI - The competence to acquire cellular desiccation tolerance is independent of seed morphological development. AB - Acquisition of desiccation tolerance and the related changes at the cellular level in wheat (Triticum aestivum cv. Priokskaya) kernels during normal development and premature drying on the ear were studied using a spin probe technique and low temperature scanning electron microscopy. During normal development, the ability of embryos to germinate after rapid drying and rehydration was acquired after completion of morphological development, which is a few days before mass maturity. The acquisition of desiccation tolerance, as assessed by germination, was associated with an upsurge in cytoplasmic viscosity, the onset of accumulation of protein and oil bodies, and the retention of membrane integrity upon dehydration/rehydration. These features were also used to assess cellular desiccation tolerance in the cases when germination could not occur. Slow premature drying was used to decouple the acquisition of cellular desiccation tolerance from morphogenesis. Upon premature drying of kernels on the ears of plants cut at 5 d after anthesis, desiccation-tolerant dwarf embryos were formed that were able to germinate. When plants were cut at earlier stages poorly developed embryos were formed that were unable to germinate, but cellular desiccation tolerance was nevertheless acquired. In such prematurely dried kernels, peripheral meristematic endosperm cells had already passed through similar physiological and ultrastructural changes associated with the acquisition of cellular desiccation tolerance. It is concluded that despite the apparent strong integration in seed development, desiccation tolerance can be acquired by the meristematic cells in the developing embryo and cambial layer of endosperm, independently of morphological development. PMID- 11432919 TI - Changes in starch content in oat (Avena sativa) shoot pulvini during the gravitropic response. AB - In order to determine if components of the signal transduction pathway are involved in starch metabolism during the gravitropic response, the effects of inhibitors of phosphoprotein phosphatases and protein kinases (OA), and calcium channel blockers (LaCl3), on gravitropic bending and starch levels in gravisensitive node/pulvini of oat shoots were examined. Among the compounds tested, okadaic acid (OA) and lanthanum chloride (LaCl3) showed the strongest inhibitory effects on the negative gravitropic curvature response in oat shoot node/pulvini. At the same time, they caused a rapid loss of starch in graviresponding pulvini based on a quantitative analysis of starch levels in the bending tissues over 48 h periods. These two compounds act initially to block the net increase in starch content that occurs during the early stages (0-9 h) in graviresponding oat shoot pulvini. As a result, starch levels drop precipitously in shoots treated with OA and LaCl3, starting at time zero of gravistimulation by reorientation. These findings suggest that protein dephosphorylation and calcium play a role in starch metabolism in oat shoot pulvini in response to a gravistimulation signal. They also indicate that the amount of starch present in the chloroplast gravisensors in oat shoot pulvini may determine the rate of upward bending in graviresponding pulvini. PMID- 11432920 TI - Quantification of insect nitrogen utilization by the venus fly trap Dionaea muscipula catching prey with highly variable isotope signatures. AB - Dionaea is a highly specialized carnivorous plant species with a unique mechanism for insect capture. The leaf is converted into an osmotically driven trap that closes when an insect triggers sensory trichomes. This study investigates the significance of insect capture for growth of Dionaea at different successional stages after a fire, under conditions where the prey is highly variable in its isotope signature. The contribution of insect-derived nitrogen (N) was estimated using the natural abundance of 15N. In contrast to previous 15N studies on carnivorous plants, the problem emerges that delta15N values of prey insects ranged between -4.47 per thousand (grasshoppers) and +7.21 per thousand (ants), a range that exceeds the delta15N values of non carnivorous reference plants (-4.2 per thousand) and soils (+3 per thousand). Thus, the isotope-mixing model used by Shearer and Kohl to estimate the amount of insect-derived N is not applicable. In a novel approach, the relationships of delta15N values of different organs with delta15N of trapping leaves were used to estimate N partitioning within the plant. It is estimated that soon after fire approximately 75% of the nitrogen is obtained from insects, regardless of plant size or developmental stage. The estimates are verified by calculating the average isotope signatures of insects from an isotope mass balance and comparing this with the average measured delta15N values of insects. It appears that for Dionaea to survive and reach the flowering stage, seedlings must first reach the 6th-leaf rosette stage, in which trap surface area nearly doubles and facilitates the capture of large insects. Large amounts of nitrogen thus made available to plants may facilitate an enhanced growth rate and the progressive production of additional large traps. Dionaea reaches a maximum abundance after fire when growth of the competing vegetation is suppressed. About 10 years after fire, when grasses and shrubs recover, Dionaea becomes overtopped by other species. This would not only reduce carbon assimilation but also the probability of catching larger prey. The amount of insect-derived nitrogen decreases to 46%, and Dionaea becomes increasingly dependent on N-supply from the soil. Competition for both light and N may cause the near disappearance of Dionaea in older stages of the fire succession. PMID- 11432921 TI - Frequencies of plasmodesmata in Allium cepa L. roots: implications for solute transport pathways. AB - Plasmodesmatal frequencies (PFs) were analysed in Allium cepa L. roots with a mature exodermis (100 mm from the tip). For all interfaces within the root, the numbers of plasmodesmata (PD) microm(-2) wall surface (Fw) were calculated from measurements of 60 walls on ultrathin sections. For tissues ranging from the epidermis up to the stelar parenchyma, the frequencies were also expressed as total PD numbers mm(-1) root length (Fn), which is most instructive for considering the radial transport of ions and photosynthates (because the tissues were arranged in concentric cylinders). The Fn values were constantly high at the interfaces of exodermis-central cortex, central cortex-endodermis and endodermis pericycle (4.05x10(5), 5.13x10(5), and 5.64x10(5), respectively). If the plasmodesmata are functional, a considerable symplastic transport pathway exists between the exodermis and pericycle. Two interfaces had especially low PFs: epidermis-exodermis (Fn=8.96x10(4)) and pericycle-stelar parenchyma (Fn=6.44x10(4)). This suggests that there is significant membrane transport across the interface of epidermis-exodermis (through short cells) and direct transfer of ions from pericycle to protoxylem vessels. In the phloem, the highest PF was detected at the metaphloem sieve element-companion cell interface (Fw=0.42), and all other interfaces had much lower PFs (around 0.10). In the pericycle, the radial walls had a high PF (Fw=0.75), a feature that could permit lateral circulation of solutes, thus facilitating ion (inward) and photosynthate (outward) delivery. PMID- 11432922 TI - Alterations in carbon and nitrogen metabolism induced by water deficit in the stems and leaves of Lupinus albus L. AB - Water deficit (WD) in Lupinus albus L. brings about tissue-specific responses that are dependent on stress intensity. Carbohydrate metabolism is very sensitive to changes in plant water status. Six days from withholding water (DAW), sucrose, glucose and fructose levels of the leaf blade had already increased over 5-fold, and the activities of SS and INV(A) had increased c. 1.5-2 times. From 9 DAW on, when stress intensity was more pronounced, these effects were reversed with fructose and glucose concentrations as well as INV(A) activity dropping in parallel. The stem (specifically the stele) responded to the stress intensification with striking increases in the concentration of sugars, N and S, and in the induction of thaumatin-like-protein and an increase in chitinase and peroxidase. At 13 DAW, the plants lost most of the leaves but on rewatering they fully recovered. Thus, the observed changes appear to contribute to a general mechanism of survival under drought, the stem playing a key role in that process. PMID- 11432923 TI - Over-expression of cytosolic glutamine synthetase increases photosynthesis and growth at low nitrogen concentrations. AB - Nitrogen, which is a major limiting nutrient for plant growth, is assimilated as ammonium by the concerted action of glutamine synthetase (GS) and glutamate synthase (GOGAT). GS catalyses the critical incorporation of inorganic ammonium into the amino acid glutamine. Two types of GS isozymes, located in the cytosol (GS1) and in the chloroplast (GS2) have been identified in plants. Tobacco (Nicotiana tabacum) transformants, over-expressing GS1 driven by the constitutive CaMV 35S promoter were analysed. GS in leaves of GS-5 and GS-8 plants was up regulated, at the level of RNA and proteins. These transgenic plants had six times higher leaf GS activity than controls. Under optimum nitrogen fertilization conditions there was no effect of GS over-expression on photosynthesis or growth. However, under nitrogen starvation the GS transgenics had c. 70% higher shoot and c. 100% greater root dry weight as well as 50% more leaf area than low nitrogen controls. This was achieved by the maintenance of photosynthesis at rates indistinguishable from plants under high nitrogen, while photosynthesis in control plants was inhibited by 40-50% by nitrogen deprivation. It was demonstrated that manipulation of GS activity has the potential to maintain crop photosynthetic productivity while reducing nitrogen fertilization and the concomitant pollution. PMID- 11432924 TI - Low sink demand limits photosynthesis under P(i) deficiency. AB - The role of the demand for carbon assimilates (the 'sink') in regulating photosynthetic carbon assimilation (Pn: the 'source') in response to phosphate (P(i)) deficiency was examined in tobacco (Nicotiana tabacum L.). P(i) supply was maintained or withdrawn from plants, and in both treatments the source/sink ratio was decreased in some plants by darkening all but two source leaves (partially darkened plants). The remaining plants were kept fully illuminated. P(i) sufficient plants showed little variation in rate of Pn, amounts of P(i) or phosphorylated intermediates. Withdrawal of P(i) decreased Pn by 75% under the growing conditions and at both low and high internal CO2 concentration. Concomitantly, P(i), phosphorylated intermediates and ATP contents decreased and starch increased. RuBP and activity of phosphoribulokinase closely matched the changes in Pn, but Rubisco activity remained high. Partial darkening P(i) deficient plants delayed the loss of photosynthetic activity; Rubisco and phosphoribulokinase activities and amounts of sucrose and metabolites, particularly RuBP and G6P, were higher than in fully illuminated Pi-deficient plants. Rates of sucrose export from leaves were more than 2-fold greater than in fully illuminated P(i)-deficient plants. Greater sucrose synthesis, facilitated by increased G6P content, an activator of SPS, would recycle P(i) from the cytosol back to the chloroplast, maintaining ATP, RuBP and hence Pn. It is concluded that low sink strength imposes the primary limitation on photosynthesis in P(i)-deficient plants which restricts sucrose export and sucrose synthesis imposing an end-product synthesis limitation of photosynthesis. PMID- 11432925 TI - Phytosiderophore release in Aegilops tauschii and Triticum species under zinc and iron deficiencies. AB - Using three diploid (Triticum monococcum, AA), three tetraploid (Triticum turgidum, BBAA), two hexaploid (Triticum aestivum and Triticum compactum, BBAADD) wheats and two Aegilops tauschii (DD) genotypes, experiments were carried out under controlled environmental conditions in nutrient solution (i) to study the relationships between the rates of phytosiderophore (PS) release from the roots and the tolerance of diploid, tetraploid, and hexaploid wheats and AE: tauschii to zinc (Zn) and iron (Fe) deficiencies, and (ii) to assess the role of different genomes in PS release from roots under different regimes of Zn and Fe supply. Phytosiderophores released from roots were determined both by measurement of Cu mobilized from a Cu-loaded resin and identification by using HPLC analysis. Compared to tetraploid wheats, diploid and hexaploid wheats were less affected by Zn deficiency as judged from the severity of leaf symptoms. Aegilops tauschii showed very slight Zn deficiency symptoms possibly due to its slower growth rate. Under Fe-deficient conditions, all wheat genotypes used were similarly chlorotic; however, development of chlorosis was first observed in tetraploid wheats. Correlation between PS release rate determined by Cu-mobilization test and HPLC analysis was highly significant. According to HPLC analysis, all genotypes of Triticum and AE: tauschii species released only one PS, 2'-deoxymugineic acid, both under Fe and Zn deficiency. Under Zn deficiency, rates of PS release in tetraploid wheats averaged 1 micromol x (30 plants)(-1) x (3 h)(-1), while in hexaploid wheats rate of PS release was around 14 micromol x (30 plants)(-1) x (3 h)(-1). Diploid wheats and AE: tauschii accessions behaved similarly in their capacity to release PS and intermediate between tetraploid and hexaploid wheats regarding the PS release capacity. All Triticum and Aegilops species released more PS under Fe than Zn deficiency, particularly when the rate of PS release was expressed per unit dry weight of roots. On average, the rates of PS release under Fe deficiency were 3.0, 5.7, 8.4, and 16 micromol x (30 plants)(-1) x (3 h)(-1) for AE: tauschii, diploid, tetraploid and hexaploid wheats, respectively. The results of the present study show that the PS release mechanism in wheat is expressed effectively when three genomes, A, B and D, come together, indicating complementary action of the corresponding genes from A, B and D genomes to activate biosynthesis and release of PS. PMID- 11432926 TI - Differential antioxidative responses to cadmium in roots and leaves of pea (Pisum sativum L. cv. Azad). AB - Pea (Pisum sativum L. cv. Azad) plants exposed to 4 and 40 microM of Cd for 7 d in hydroponic culture were analysed with reference to the distribution of metal, the accumulation of biomass and the metal's effects on antioxidants and antioxidative enzymes in roots and leaves. Cd-induced a decrease in plant biomass. The maximum accumulation of Cd occurred in roots followed by stems and leaves. An enhanced level of lipid peroxidation and an increased tissue concentration of hydrogen peroxide (H2O2) in both roots and leaves indicated that Cd caused oxidative stress in pea plants. Roots and leaves of pea plants responded differently to Cd with reference to the induction of enhanced activities of most of the enzymes monitored in the present study. These differential responses to Cd were further found to be associated with levels of Cd to which the plants were exposed. Cd-induced enhancement in superoxide dismutase (SOD) activity was more at 40 microM than at 4 microM in leaves. While catalase (CAT) prominently increased in leaves both at 4 and 40 microM Cd, ascorbate peroxidase (APX) showed maximum stimulation at 40 microM Cd in roots. Enhancement in glutathione reductase (GR) activity was also more at 40 microM than at 4 microM Cd in roots. While glutathione peroxidase (GPOX) activity decreased in roots and remained almost unmodified in leaves, glutathione S transferase (GST) showed pronounced stimulation in both roots and leaves of pea plants exposed to 40 microM Cd. Increased activities of antioxidative enzymes in Cd-treated plants suggest that they have some additive function in the mechanism of metal tolerance in pea plants. PMID- 11432927 TI - Two APETALA2-like genes of Picea abies are differentially expressed during development. AB - The EREBP/AP2 gene family codes for plant specific transcription factors. The first two gymnosperm genes of this family were isolated from Picea abies and shown to be structurally related, but not orthologous, to the angiosperm AP2-like genes. The two P. abies genes are differentially expressed in different organs and may be important developmental regulators. PMID- 11432928 TI - Leaf senescence in rice plants: cloning and characterization of senescence up regulated genes. AB - To identify senescence-associated genes (SAGs) in rice leaves, senescence was induced by transferring rice seedlings into darkness. Senescence up-regulated cDNAs were obtained by PCR-based subtractive hybridization. Among 14 SAG clones characterized, 11 were found to be associated with both dark-induced and natural leaf senescence. Three clones were associated only with dark-induced leaf senescence. The possible physiological roles of these SAGs during rice leaf senescence are discussed. PMID- 11432929 TI - Adaptation to long-term cold-girdling in genotypes of common bean (Phaseolus vulgaris L.). AB - The effect of long-term cold-girdling on phloem transport and resource allocation in whole plants of common bean is described. Wide differences were found between genotypes, with some maintaining translocation when cold-girdled. This provides evidence to support passive phloem transport. The possibilities that cold girdling may physically block transport and/or disrupt root-shoot signalling are discussed. PMID- 11432930 TI - Water stress can induce quiescence in newly-germinated onion (Allium cepa L.) seedlings. AB - The effect of water stress on the early seedling growth of onions was studied by placing newly-germinated seedlings in vermiculite equilibrated at different water potentials. Roots and shoots elongated more at -0.29 than at -0.64 MPa, but did not elongate at -1.66 MPa. However, roots and shoots of seedlings that had been incubated in vermiculite at -1.66 MPa for up to 35 d resumed elongation when subsequently placed on wet filter boards. This suggests that water stress can induce quiescence in newly-germinated seedlings. PMID- 11432931 TI - Factors influencing Agrobacterium-mediated transient expression of uidA in wheat inflorescence tissue. AB - A critical step in the development of Agrobacterium tumifaciens-mediated transformation is the establishment of optimal conditions for T-DNA delivery into tissue from which whole plants can be regenerated. The efficient transformation of inflorescence tissue from 'Baldus', a commercial wheat variety, using the Agrobacterium strain AGLI harbouring the binary vector pAL156 is reported here. The effects of various factors on delivery and the transient expression of the uidA gene were studied including the duration of preculture, vacuum infiltration, the effect of sonication treatments, and Agrobacterium cell density. Optimal T DNA delivery (as measured by uidA activity) was obtained from inflorescence tissues precultured for 21 d and sonicated. Increasing Agrobacterium cell density, the duration of inoculation/co-cultivation, and vacuum pressure, up to a threshold, increased uidA expression. The investigation of factors that influence T-DNA delivery is an important first step in the utilization of Agrobacterium in the transformation of immature wheat inflorescence tissue. PMID- 11432932 TI - Drought differentially affects expression of a PR-10 protein, in needles of maritime pine (Pinus pinaster Ait.) seedlings. AB - A full length cDNA encoding a PR-10 protein was isolated from maritime pine drought-stressed seedlings. The predicted protein contained 150 amino acids, has a molecular mass of 16.7 kDa and an isoelectric point of 5.28. The transcript level of PR-10 displayed a transient accumulation in needles of drought-stressed plants, and was not detectable in root and stem tissues. PMID- 11432933 TI - The pathway of biosynthesis of abscisic acid in vascular plants: a review of the present state of knowledge of ABA biosynthesis. AB - The pathway of biosynthesis of abscisic acid (ABA) can be considered to comprise three stages: (i) early reactions in which small phosphorylated intermediates are assembled as precursors of (ii) intermediate reactions which begin with the formation of the uncyclized C40 carotenoid phytoene and end with the cleavage of 9'-cis-neoxanthin (iii) to form xanthoxal, the C15 skeleton of ABA. The final phase comprising C15 intermediates is not yet completely defined, but the evidence suggests that xanthoxal is first oxidized to xanthoxic acid by a molybdenum-containing aldehyde oxidase and this is defective in the aba3 mutant of Arabidopsis and present in a 1-fold acetone precipitate of bean leaf proteins. This oxidation precludes the involvement of AB-aldehyde as an intermediate. The oxidation of the 4'-hydroxyl group to the ketone and the isomerization of the 1',2'-epoxy group to the 1'-hydroxy-2'-ene may be brought about by one enzyme which is defective in the aba2 mutant and is present in the 3-fold acetone fraction of bean leaves. Isopentenyl diphosphate (IPP) is now known to be derived by the pyruvate-triose (Methyl Erythritol Phosphate, MEP) pathway in chloroplasts. (14C)IPP is incorporated into ABA by washed, intact chloroplasts of spinach leaves, but (14C)mevalonate is not, consequently, all three phases of biosynthesis of ABA occur within chloroplasts. The incorporation of labelled mevalonate into ABA by avocado fruit and orange peel is interpreted as uptake of IPP made in the cytoplasm, where it is the normal precursor of sterols, and incorporated into carotenoids after uptake by a carrier in the chloroplast envelope. An alternative bypass pathway becomes more important in aldehyde oxidase mutants, which may explain why so many wilty mutants have been found with this defect. The C-1 alcohol group is oxidized, possibly by a mono-oxygenase, to give the C-1 carboxyl of ABA. The 2-cis double bond of ABA is essential for its biological activity but it is not known how the relevant trans bond in neoxanthin is isomerized. PMID- 11432934 TI - Antibodies to assess phosphorylation of spinach leaf nitrate reductase on serine 543 and its binding to 14-3-3 proteins. AB - To monitor site-specific phosphorylation of spinach leaf nitrate reductase (NR) and binding of the enzyme to 14-3-3 proteins, serum antibodies were raised that select for either serine 543 phospho- or dephospho-NR. The dephospho-specific antibodies blocked NR phosphorylation on serine 543. The phospho-specific antibodies prevented NR binding to 14-3-3s, NR inhibition by 14-3-3s, NR dephosphorylation on serine 543, and did not precipitate 14-3-3s together with NR. Together, this confirms that 14-3-3s bind to NR at hinge 1 after it has been phosphorylated on serine 543. The amounts of individual NR forms were determined in leaf extracts by immunoblotting and immunoprecipitation. The phosphorylation state of NR on serine 543 increased 2-3-fold in leaves upon a light/ dark transition. Before the transition, one-third of NR was already phosphorylated on serine 543 but was not bound to 14-3-3s. Phosphorylation of serine 543 seems not to be enough to bind to 14-3-3s in leaves. PMID- 11432935 TI - Through pore diameter in the cell wall of Chara corallina. AB - Determination of pore size of the cell wall of Chara corallina has been made by using the polyethylene glycol (PEG) series as the hydrophilic probing molecules. In these experiments, the polydispersity of commercial preparation of PEGs was allowed for. The mass share (gamma(p)) of polyethylene glycol preparation fractions penetrating through the pores was determined using a cellular 'ghost', i.e. fragments of internodal cell walls filled with a 25% solution of non penetrating PEG 6000 and tied up at the ends. In water, such a 'ghost' developed a hydrostatic pressure close to the cell turgor which persisted for several days. The determination of gamma(p), for polydisperse polyethylene glycols with different average molecular mass (M) was calculated from the degree of pressure restoration after water was replaced by a 5-10% polymer solution. Pressure was recorded using a dynamometer, which measures, in the quasi-isometric mode, the force necessary for the partial compression of the 'ghost' in its small fragment. By utilizing the data on the distribution of PEG 1000, 1450, 2000, and 3350 fractions over molecular mass (M), it was found that gamma(p), for these polyethylene glycols corresponded to the upper limit of ML=800-1100 D (hydrodynamic radius of molecules, r(h)=0.85-1.05 nm). Thus, the effective diameter of the pores in the cell wall of Chara did not exceed 2.1 nm. PMID- 11432936 TI - Anoxic stress leads to hydrogen peroxide formation in plant cells. AB - Hydrogen peroxide (H2O2) was detected cytochemically in plant tissues during anoxia and re-oxygenation by transmission electron microscopy using its reaction with cerium chloride to produce electron dense precipitates of cerium perhydroxides. Anoxia-tolerant yellow flag iris (Iris pseudacorus) and rice (Oryza sativa), and anoxia-intolerant wheat (Triticum aestivum) and garden iris (Iris germanica) were used in the experiments. In all plants tested, anoxia and re-oxygenation increased H2O2 in plasma membranes and the apoplast. In the anoxia tolerant species the response was delayed in time, and in highly tolerant I. pseudacorus plasma membrane associated H2O2 was detected only after 45 d of oxygen deprivation. Quantification of cerium precipitates showed a statistically significant increase in the amount of H2O2 caused by anoxia in wheat root meristematic tissue, but not in the anoxia-tolerant I. pseudacorus rhizome parenchyma. Formation of H2O2 under anoxia is considered mainly an enzymatic process (confirmed by an enzyme inhibition analysis) and is due to the trace amount of dissolved oxygen (below 10(-5) M) present in the experimental system. The data suggest oxidative stress is an integral part of oxygen deprivation stress, and emphasize the importance of the apoplast and plasma membrane in the development of the anoxic stress response. PMID- 11432937 TI - Subcellular distribution and kinetic properties of cytosolic and non-cytosolic hexokinases in maize seedling roots: implications for hexose phosphorylation. AB - Hexose phosphorylation by hexokinases plays an important role in glycolysis, biosynthesis and control of sugar-modulated genes. Several cytosolic hexokinase and fructokinase isoforms have been characterized and organelle-bound hexokinases have also been detected in higher plants. In this study a hexokinase activity is described that is inhibited by ADP (K(i)=30 microM) and mannoheptulose (K(i) congruent with 300 microM) in non-cytosolic fractions (mitochondria, Golgi apparatus and microsomes) obtained from preparations of seedling roots of maize (Zea mays L.). The catalytic efficiency (Vmax/Km) for both ATP and glucose in all non-cytosolic hexokinase fractions is more than one order of magnitude higher than that of cytosolic hexokinase and fructokinases. Low (30%) or no ADP and mannoheptulose inhibition is observed with hexokinase and fructokinase activities derived from the cytosolic compartment obtained after ion exchange and affinity chromatography. The soluble fructokinase (FK) shows fructose cooperativity (Hill n>2). The Vmax/Km ratio is about 3-fold higher for ATP than for other NTPs and no difference for hexose phosphorylation efficiencies is found between cytosolic hexokinase and fructokinase isoforms (FK1, FK2) with ATP as substrate. The K(i) for fructose inhibition is 2 mM for FK1 and 25 mM for FK2. The data indicate that low energy-charge and glucose analogues preferentially inhibit the membrane-bound hexokinases possibly involved in sugar-sensing, but not the cytosolic hexokinases and fructokinases. PMID- 11432938 TI - Dormancy and proliferation in Saccharum officinarumxS. spontaneum hybrids which differ in the number of the introgressed S. spontaneum chromosomes. AB - Proliferating cells remain transiently blocked at different cycle compartments until specific stressors are removed or until the cells become adapted to their presence. This paper investigates the efficiency of cycle blocks in three sugarcane hybrids with the full noble cane (Saccharum officinarum) genome (2n=8x=80) but differing in the number of introgressed S. spontaneum (2n=8x=64) chromosomes. The My5514, B42231 and C236-51 cultivars possess 20, 30 and 40 additional S. spontaneum chromosomes, respectively. Flow cytometry showed that over 90% of cells were accumulated with a 2C DNA content in their dormant primordia. The presence of S. spontaneum chromosomes decreased the low stringency of the 4C block. The greater the number of these chromosomes, the lower was the number of quiescent cells with a 4C DNA content (P<0.05). Shortly after stimulation of the primordia (85% relative humidity and 30 degrees C), i.e. in the 2 mm long roots, a negative correlation was found between the number of introgressed S. spontaneum chromosomes and the frequency of cells undergoing replication and mitosis. On the other hand, when roots were already proliferating under steady-state conditions (15 mm long roots) the more S. spontaneum chromosomes the cells possessed, the longer the relative time it took for all chromosomes to replicate and segregate, and the longer the relative time they spent in G(2), with the 4C DNA content. The presence of S. spontaneum chromosomes seems to be recognized by these proliferating cells as a stressor which preferentially activates checkpoint pathways operating at the second half of the cycle, but not at its onset. PMID- 11432939 TI - Mapping of QTLs associated with cytosolic glutamine synthetase and NADH-glutamate synthase in rice (Oryza sativa L.). AB - Ninety-eight backcross inbred lines (BC1F6) developed between Nipponbare, a japonica rice, and Kasalath, an indica rice were employed to detect putative quantitative trait loci (QTLs) associated with the contents of cytosolic glutamine synthetase (GS1; EC 6.3.1.2) and NADH-glutamate synthase (NADH-GOGAT; EC 1.4.1.14) in leaves. Immunoblotting analyses showed transgressive segregations toward lower or greater contents of these enzyme proteins in these backcross inbred lines. Seven chromosomal QTL regions for GS1 protein content and six for NADH-GOGAT protein content were detected. Some of these QTLs were located in QTL regions for various biochemical and physiological traits affected by nitrogen recycling. These findings suggested that the variation in GS1 and NADH-GOGAT protein contents in this population is related to the changes in the rate of nitrogen recycling from senescing organs to developing organs, leading to changes in these physiological traits. Furthermore, a structural gene for GS1 was mapped between two RFLP markers, C560 and C1408, on chromosome 2 and co-located in the QTL region for one-spikelet weight. A QTL region for NADH-GOGAT protein content was detected at the position mapped for the NADH-GOGAT structural gene on chromosome 1. A QTL region for soluble protein content in developing leaves was also detected in this region. Although fine mapping is required to identify individual genes in the future, QTL analysis could be a useful post-genomic tool to study the gene functions for regulation of nitrogen recycling in rice. PMID- 11432940 TI - Ca2+-dependent and Ca2+-independent excretion modes of salicylic acid in tobacco cell suspension culture. AB - 14C-salicylic acid (SA) was used to monitor SA metabolism and its regulation in tobacco cell suspension culture. Two SA concentrations (20 microM and 200 microM) were used for comparison. SA was quickly taken up in both treatments, and the 200 microM-treated cells absorbed approximately 15 times that of 20 microM-treated cells within 5 min. More than 85% and 50% of the absorbed SA were excreted in free form to the culture medium within 5 h from cells treated with 200 microM and 20 microM SA, respectively. SA excretion was significantly inhibited by EGTA and the inhibition could be reversed by the addition of exogenous Ca2+ to the culture medium in the 200 microM SA treatment. However, EGTA had little or no effect on SA excretion in the 20 microM SA treatment. The data suggest that tobacco suspension-cultured cells may contain both Ca2+-dependent and Ca2+-independent pathways for SA excretion. Reduced glutathione (an active oxygen species scavenger), staurosporine (a protein kinase inhibitor), and cycloheximide (an inhibitor of de novo protein synthesis) also blocked intracellular SA excretion to the culture medium in the 200 microM but not in the 20 microM SA treatment. These data support the existence of alternative SA excretion pathways in tobacco suspension-cultured cells. Tobacco cells may use both Ca2+-dependent and Ca2+ independent excretion pathways to cope with different intracellular SA status, and the pathway influenced by EGTA, reduced glutathione, staurosporine, and cycloheximide is activated by SA at 200 microM, but not at 20 microM. PMID- 11432941 TI - Nuclear fusion leads to chromosome doubling during mannitol pretreatment of barley (Hordeum vulgare L.) microspores. AB - A cytological study of barley microspores during pretreatment of the uninucleate stage to the early culture stage was conducted utilizing six genotypes. Among the three main pretreatments investigated, microspores completed the first mitotic division during 28 d cold pretreatment of spikes, with or without leaf sheath attached, and during 0.3 M mannitol pretreatment of anthers at 25 degrees C. However, during a 4 d pretreatment in 0.3 M mannitol at 4 degrees C this first mitotic division was blocked or delayed and subsequently most often occurred during the first day on culture medium. The first mitotic division of most microspores pretreated in 0.3 M mannitol was mostly symmetrical (55-60%), whereas it was asymmetric (94%) during the 28 d cold pretreatment of spikes. Following the first mitotic division during the mannitol pretreatment at 25 degrees C, closely associated daughter nuclei often appeared to fuse via membrane coalescence, leading to a high frequency of large uninucleate microspores. Based upon nuclear size, the frequencies of fused uninucleate microspores in genotypes GBC 778, GBC 777 and Igri were estimated to be 87%, 54% and 75%, respectively, after a 4 d mannitol pretreatment at 25 degrees C. Chromosome numbers in dividing nuclei and relative densitometry measurements of nuclear DNA in microspores from cv. Igri confirmed the apparent fused nature of large nuclei in uninucleate microspores. The high frequency of fused nuclei indicates that nuclear fusion occurred between both symmetric and asymmetric nuclei. Microspores of cv. Igri cultured on filter paper following three different pretreatments provided an average of about 12 000 embryo-like structures (ELS) per plate. In samples, 85 97% of these ELS regenerated green shoots. The frequency of doubled haploids (74 83%) following all pretreatments was similar to the frequencies of fused nuclei. The pretreatment of spikes in 0.3 M mannitol at 4 degrees C for 4 d is preferred as it appears to provide genotype independent induction and suspension of nuclear division, as well as regenerating green plants in a shorter time than cold alone. PMID- 11432942 TI - The cloning and characterization of alpha-galactosidase present during and following germination of tomato (Lycopersicon esculentum Mill.) seed. AB - alpha-Galactosidase (EC 3.2.1.22) is present in the embryo, micropylar and lateral endosperm of seeds of tomato during and following germination. Its activity is unchanged even when germination of the seeds is prevented by an osmoticum. It is also present in the developing and mature dry seed. A cDNA clone for tomato seed alpha-galactosidase (LeaGal) has been isolated and the characteristics of the protein deduced; the predicted molecular mass of the mature enzyme is 39.8 kDa, with a pI of 4.91. The tomato alpha-galactosidase has a high homology (>62%) at the amino acid level with that of other plant alpha galactosidases. A hydrophobic signal peptide region is identified which is indicative that the enzyme enters the lumen of the endoplasmic reticulum during its translation, prior to its export to the protein body or cell wall, the presumed sites of its substrates. Using amino acid alignment and phylogenetic analysis, key amino acids have been identified, and relationships to other alpha galactosidases inferred. Southern hybridization analyses show that the enzyme is derived from a single gene (for which a partial sequence has been obtained) and yet there are at least three different isoforms within the seed; post translational modifications are thus presumed to occur. From Northern hybridization studies it is evident that alpha-galactosidase transcripts are present in the lateral and micropylar endosperm during and following germination, and also to a lesser extent in the embryo. PMID- 11432943 TI - Tobacco plants that lack expression of functional nitrate reductase in roots show changes in growth rates and metabolite accumulation. AB - When tobacco is provided with a high nitrate supply, only a small amount of the nitrate taken up by the roots is immediately assimilated inside the roots, while the majority is transported to the leaves where it is reduced to ammonium. To elucidate the importance of root nitrate assimilation, tobacco plants have been engineered that showed no detectable nitrate reductase activity in the roots. These plants expressed the nitrate reductase structural gene nia2 under control of the leaf-specific potato promoter ST-LS1 in the nitrate reductase-mutant Nia30 of Nicotiana tabacum. Homozygous T2-transformants grown in sand or hydroponics with 5.1 mM nitrate had approximately 55-70% of wild-type nitrate reductase acivity in leaves, but lacked nitrate reductase acivity in roots. These plants showed a retarded growth as compared with wild-type plants. The activation state of nitrate reductase was unchanged; however, diurnal variation of nitrate reductase acivity was not as pronounced as in wild-type plants. The transformants had higher levels of nitrate in the leaves and reduced amounts of glutamine both in leaves and roots, while roots showed higher levels of hexoses (3-fold) and sucrose (10-fold). It may be concluded that the loss of nitrate reductase acivity in the roots changes the allocation of reduced nitrogen compounds and sugars in the plant. These plants will be a useful tool for laboratories studying nitrate assimilation and its interactions with carbon metabolism. PMID- 11432944 TI - The elongation rate at the base of a maize leaf shows an invariant pattern during both the steady-state elongation and the establishment of the elongation zone. AB - Spatial and temporal analyses of elongation and cell length of monocotyledon leaves have most often been performed during the period when leaves are visible and elongate at a constant rate (steady-state). In the present study, the focus was on the earlier stages, during the establishment of the elongation zone. Regardless of leaf development stage, the segment located between 0 and 35 mm from the leaf insertion point had a relative elongation rate that increased with distance from insertion point ('accelerating zone') while the segment located further than 35 mm had a relative elongation rate that decreased ('decelerating zone'). This stable pattern held for both young, non-emerged leaves, where it was restricted to the portion corresponding to the length of the blade, and for leaves during steady-state elongation. In the same way, the profile of cell length was essentially the same during early development and during steady-state elongation. The results of a temporal analysis of whole-leaf elongation rate, carried out in the field and in the greenhouse at different light intensities were consistent with a time-invariant pattern of elongation. Whole-leaf relative elongation rate increased with time until the leaf reached 30-40 mm length (although at different leaf ages depending on conditions), and declined afterwards. These results suggest that the patterns governing the elongation rate of a sector of a maize leaf are independent of the leaf developmental stage but depend on sector position only. PMID- 11432945 TI - Transport and accumulation rates of abscisic acid and aldehyde oxidase activity in Pisum sativum L. in response to suboptimal growth conditions. AB - Pea plants (Pisum sativum L.) grown initially in nutrient solutions with adequate nitrogen supply (4 mM NO3-) were transferred to solutions containing salt (50 or 100 mM NaCl), ammonium (4 mM) or a low nitrogen supply (0.4 mM NO3-). No changes of abscisic acid (ABA) content were found in roots of stressed pea plants 9 d after the beginning of the treatments; however, accumulation of ABA in the leaves was observed. Old leaves accumulated ABA to a higher extent than young leaves. Accumulation of ABA in leaves of ammonium-fed plants and plants grown under low nitrogen supply occurred in the absence of both increased ABA xylem loading rate and enhanced aldehyde oxidase (AO, EC 1.2.3.1) activity in roots. Enhanced leaf AO activity was observed in all treatments, with the highest increase in old leaves. Among the three AO isoforms (AO-1, AO-2 and AO-3) detected in extracts of pea leaves, the lowest one AO-3 (highest mobility in the gel) correlated with ABA production and showed the highest increment in response to the treatments. The increase of AO activity detected in leaves after 2 weeks of stress application was less prominent than after 9 d, suggesting a transient enhancement of ABA production following the onset of stress. An increase of ABA xylem loading rate as well as AO root activity 4 d and 9 d after application of the treatments was observed only in salt-treated plants followed by a decrease after 14 d in 100 mM NaCl. Decreased cytokinin (trans-zeatin riboside) delivery rate into the xylem sap was observed in all treatments. The role of abscisic acid and cytokinins as positive and negative growth signals, as well as the involvement of root generated ABA on ABA accumulation in leaves is discussed. PMID- 11432946 TI - Plant growth and cation composition of two cultivars of spring wheat (Triticum aestivum L.) differing in P uptake efficiency. AB - Phosphorus (P)-zinc (Zn) interactions were investigated in two wheat cultivars (Brookton versus Krichauff) differing in P uptake efficiency. The experiment was done in a growth chamber. Rock phosphate (RP) or CaHPO4 (CaP) were used as P sources, and ammonium nitrate (AN) or nitrate only (NO) were used as nitrogen sources. Two Zn levels were used, 0.22 mg x kg(-1) (LZ) and 2.2 mg ZnSO4.7H2O x kg(-1) (HZ), respectively. P availability significantly affected plant biomass production, but Zn supply had little effect. Plants fed ammonium nitrate had significantly lower concentrations of cations than those fed nitrate only. Cultivar Brookton (with higher P uptake efficiency) consistently had lower concentrations of cations than cv. Krichauff (with low P uptake efficiency) under limited P supply. The differences in concentrations of cations increased with the decrease in P availability, but were not affected by Zn supply. The ratio of potassium in roots to shoots of cultivar Brookton was always higher than in cultivar Krichauff. Based on these findings, it is postulated that the lower concentrations of cations in cultivar Brookton are related to root exudation of organic anions, and a conceptual model is established to describe the regulation of root exudation of organic anions and concentrations of cations. PMID- 11432947 TI - Diurnal changes in nitrogen assimilation of tobacco roots. AB - To gain an insight into the diurnal changes of nitrogen assimilation in roots the in vitro activities of cytosolic and plasma membrane-bound nitrate reductase (EC 1.6.6.1), nitrite reductase (EC 1.7.7.1) and cytosolic and plastidic glutamine synthetase (EC 6.3.1.2) were studied. Simultaneously, changes in the contents of total protein, nitrate, nitrite, and ammonium were followed. Roots of intact tobacco plants (Nicotiana tabacum cv. Samsun) were extracted every 3 h during a diurnal cycle. Nitrate reductase, nitrite reductase and glutamine synthetase were active throughout the day-night cycle. Two temporarily distinct peaks of nitrate reductase were detected: during the day a peak of soluble nitrate reductase in the cytosol, in the dark phase a peak of plasma membrane-bound nitrate reductase in the apoplast. The total activities of nitrate reduction were similar by day and night. High activities of nitrite reductase prevented the accumulation of toxic amounts of nitrite throughout the entire diurnal cycle. The resulting ammonium was assimilated by cytosolic glutamine synthetase whose two activity peaks, one in the light period and one in the dark, closely followed those of nitrate reductase. The contribution of plastidic glutamine synthetase was negligible. These results strongly indicate that nitrate assimilation in roots takes place at similar rates day and night and is thus differently regulated from that in leaves. PMID- 11432948 TI - Extensiometric determination of the rheological properties of the epidermis of growing tomato fruit. AB - This paper examines the rheological properties of the fruit epidermis of tomato (Lycopersicon esculentum L.). This research was conducted because previous work had demonstrated that the rate of tomato fruit growth is determined by the interaction of tissue pressure and epidermal properties. A constant-load (or 'creep') extensiometer was employed in these experiments and the results interpreted using a model which describes creep retardation using a limited number of rheological elements, one of which appears analogous to plant growth and is of similar magnitude to fruit growth rate in vivo. The effects of pH, applied force and boiling upon the individual components of the model have been examined and indicate that several elements are strongly pH-dependent and that this dependency is eliminated by boiling. These results suggest that enzyme activity (plausibly that of one or more expansins) reduces the viscosity of the cell wall over a wide range of time scales. Further consideration of the creep of tomato epidermis in terms of models developed to describe the behaviour of artificial polymers suggests that the types of molecular event described by each rheological element can tentatively be identified and that pH-dependent enzyme activity facilitates both conformer rotation and macromolecular movement within the plant cell wall. These interpretations ascribe considerable importance to the time scale over which creep occurs. PMID- 11432949 TI - Stomatal oscillations at small apertures: indications for a fundamental insufficiency of stomatal feedback-control inherent in the stomatal turgor mechanism. AB - Continuous measurements of stomatal aperture simultaneously with gas exchange during periods of stomatal oscillations are reported for the first time. Measurements were performed in the field on attached leaves of undisturbed Sambucus nigra L. plants which were subjected to step-wise increases of PPFD. Oscillations only occurred when stomatal apertures were small under high water vapour mole fraction difference between leaf and atmosphere (DeltaW). They consisted of periodically repeated opening movements transiently leading to very small apertures. Measurements of the area of the stomatal complex in parallel to the determination of aperture were used to record volume changes of guard cells even if stomata were closed. Stomatal opening upon a light stimulus required an antecedent guard cell swelling before a slit occurred. After opening of the slit the guard cells again began to shrink which, with some delay, led to complete closure. Opening and closing were rhythmically repeated. The time-lag until initial opening was different for each individual stoma. This led to counteracting movements of closely adjacent stomata. The tendency to oscillate at small apertures is interpreted as being a failure of smoothly damped feedback regulation at the point of stomatal opening: Volume changes are ineffective for transpiration if stomata are still closed; however, at the point of initial opening transpiration rate rises steeply. This discontinuity together with the rather long time constants inherent in the stomatal turgor mechanism makes oscillatory overshooting responses likely if at high DeltaW the 'nominal value' of gas exchange demands a small aperture. PMID- 11432950 TI - Uptake of glycine by non-mycorrhizal Lolium perenne. AB - Plants of Lolium perenne were grown in sterile solution culture. 15N-labelled glycine (Gly) coupled with gas chromatograph mass spectrometry was used to prove that non-mycorrhizal plants of L. perenne are capable of acquiring N in the form of intact Gly. It was estimated that a minimum of 80% of Gly-N uptake, over a 3 h period, was as intact Gly, though possible processes resulting in deviation from this estimate are discussed. The relative incorporation of 15N derived from Gly uptake into serine (Ser) compared with other amino acids in the root amino acid pool suggested the enzyme serine:glyoxylate aminotransferase was at least partly responsible for the synthesis of Ser from Gly. Defoliation was shown to reduce Gly uptake by L. perenne. The addition of either 25 mol x m(-3) sucrose or 50 mol x m(-3) glucose to the uptake solution of defoliated plants increased Gly-N uptake compared with both defoliated plants without sugars and with undefoliated plants. Addition of a glucose analogue, 3-O-methyl-D-glucopyranose, that is absorbed but not metabolized by plants, did not affect Gly uptake by defoliated plants. Increasing pH from 3.5 to 9.2 caused a reduction in Gly uptake. Results of the effects of defoliation and pH are consistent with Gly uptake by L. perenne being by an energy-dependent proton symport. When either or Gly were supplied to plants at equimolar concentrations, uptake was five times greater than that of Gly at pH 6 and 13 times greater at pH 9. PMID- 11432951 TI - Limitations to photosynthesis of lettuce grown under tropical conditions: alleviation by root-zone cooling. AB - Aerial parts of lettuce plants were grown under natural tropical fluctuating ambient temperatures, but with their roots exposed to two different root-zone temperatures (RZTs): a constant 20 degrees C-RZT and a fluctuating ambient (A-) RZT from 23-40 degrees C. Plants grown at A-RZT showed lower photosynthetic CO2 assimilation (A), stomatal conductance (gs), midday leaf relative water content (RWC), and chlorophyll fluorescence ratio Fv/Fm than 20 degrees C-RZT plants on both sunny and cloudy days. Substantial midday depression of A and g(s) occurred on both sunny and cloudy days in both RZT treatments, although Fv/Fm did not vary diurnally on cloudy days. Reciprocal temperature transfer experiments investigated the occurrence and possible causes of stomatal and non-stomatal limitations of photosynthesis. For both temperature transfers, light-saturated stomatal conductance (gs sat) and photosynthetic CO2 assimilation (A(sat)) were highly correlated with each other and with midday RWC, suggesting that A was limited by water stress-mediated stomatal closure. However, prolonged growth at A RZT reduced light- and CO2-saturated photosynthetic O2 evolution (Pmax), indicating non-stomatal limitation of photosynthesis. Tight temporal coupling of leaf nitrogen content and P(max) during both temperature transfers suggested that decreased nutrient status caused this non-stomatal limitation of photosynthesis. PMID- 11432952 TI - 14-3-3 gene family in hybrid poplar and its involvement in tree defence against pathogens. AB - In ongoing investigations of the role of the signal transduction pathway in tree pathogen interactions, four complete and two partial 14-3-3 cDNAs have been isolated which are members of a gene family. Comparisons of DNA sequences reveal a high degree of identity among the cDNAs, and, in some cases, higher than 75% sequence similarity with previously published sequences. Sequence analysis at the amino acid level uncovered potential phosphorylation sites, some of which were identical among the proteins, and some of which varied. Treatment of trees with chitosan, jasmonates or by wounding of leaves, caused increases in the levels of 14-3-3 mRNA transcripts. Since jasmonates and chitosan are signal transducers of defence reactions in plants, these results suggest a possible role for 14-3-3 proteins in the pathogen defence response of deciduous trees. Effects of elicitors on transcription of the pal gene were also monitored. Pal is a well characterized, pathogen response-related gene. PMID- 11432953 TI - The role of root exudates in aluminium resistance and silicon-induced amelioration of aluminium toxicity in three varieties of maize (Zea mays L.). AB - Aluminium (Al) toxicity is widely considered to be the most important growth limiting factor for plants in strongly acid soils (pH<5.0). The inhibition of root elongation in three varieties of maize (Zea mays L. vars Clavito, HS701b and Sikuani) was followed over the first 48 h of Al treatment, and during the initial 10 h elongation was determined on an hourly basis. The silicon (Si)-induced amelioration of Al toxicity was investigated by pre-treating seedlings for 72 h in nutrient solutions with 1000 microM Si before transfer into solutions with 0, 20 or 50 microM Al (without Si). Plants were either grown in complete low ionic strength nutrient solutions (CNS) or in low salt solutions of 0.4 mM CaCl2 (LSS). In addition, the role of root exudation of organic compounds as a mechanism of Si induced alleviation of Al toxicity was investigated. Aluminium-induced inhibition of root elongation in the maize var. HS701b was observed within 1 h of Al exposure. After a lag time of at least 8 h, Si-induced alleviation of Al toxicity was observed in this variety when grown in LSS. In the Al-resistant var. Sikuani, Al-resistance was only observed after exposure to 50 microM Al, and not after exposure to 20 microM Al, suggesting that there exists a threshold Al concentration before the mechanisms of Al resistance are activated. Aluminium stimulated root exudation of oxalic acid in all three varieties, but exudate concentrations did not increase with either Al resistance or with Si pretreatment. Aluminium and Si triggered release of catechol and of the flavonoid type phenolics: catechin, and quercetin. In the Al-resistant variety, Sikuani, Al exposed plants pretreated with Si exuded up to 15 times more phenolics than those plants not pretreated with Si. The flavonoid-type phenolics, to date unconsidered, appear to play a role in the mechanism(s) of Si-induced amelioration of Al toxicity. PMID- 11432954 TI - An improved Na+-selective microelectrode for intracellular measurements in plant cells. AB - The high background K+ concentration in plant cells is a problem for intracellular measurements of Na+ using ion-selective microelectrodes. The discrimination between Na+ and K+ of the microelectrode ionophore molecule limits the usefulness of this technique. A new Na+-selective microelectrode with an ionophore incorporating a tetramethoxyethyl ester derivative of p-t-butyl calix[4]arene has been developed. Microelectrodes made with this new sensor have superior selectivity for Na+ over K+ resulting in a lower limit of detection when compared with microelectrodes made using a commercially available ionophore (ETH227). Both types of microelectrodes were insensitive to changes in ionic strength and physiological ranges of pH, but only the calixarene-based electrodes showed no protein interference. To test the suitability of the calixarene-based microelectrodes for measurements in plants, they were used to measure Na+ in epidermal cells in the zone 10-20 mm from the root apex of barley (Hordeum vulgare L.). Seedlings were grown in a nutrient solution containing 200 mM NaCl for 1-6 d. The range of intracellular Na+ activity (a(Na)) measured varied from < or =0.1 mM (limit of detection) to over 100 mM, and these values increased significantly with time. The membrane potential (E(m)) of these cells was variable, but the values became significantly more negative with time, although there was no significant correlation between E(m) and a(Na). These intracellular measurements could not be separated into distinct populations that might be representative of subcellular compartments. PMID- 11432955 TI - A new validation of the Scholander pressure chamber technique based on stem diameter variations. AB - The Scholander pressure chamber is one of the most widely used techniques for measuring plant water status. However, the technique has been the subject of recent controversies, and its validity awaits new experimental evidence. This paper presents a new test based on the analysis of the dependence on water potential difference (DeltaPsi) of stem diameter variation (DeltaD) in walnut (Juglans regia L.). The correlation between DeltaPsi and DeltaD was established (1) on transpiring potted trees, (2) on dehydrating cut branches, (3) by perfusing the xylem of branch segments with mannitol and sucrose solutions, and (4) by pressurizing segments in a pressure sleeve. The DeltaPsi was respectively assessed with a pressure chamber (1, 2), a freezing point osmometer (3) and an air pressure transducer (4). A single relationship was established between DeltaPsi (ranging from 0 to -2 MPa) and DeltaD for all the experiments. This shows that the measured changes of water potential were correlated to similar modifications of water content in the stems, irrespective of the technique used to induce these changes, and therefore validates the pressure chamber technique and confirms the occurrence of large negative pressures in the xylem of walnut branches. PMID- 11432956 TI - Ultraviolet-B exposure leads to up-regulation of senescence-associated genes in Arabidopsis thaliana. AB - Exposure to UV-B radiation resulted in a loss of chlorophyll and an increase in lipid damage in a similar manner to that induced during natural senescence. In addition, exposure to UV-B led to the induction of a number of genes associated with senescence (SAG12, 13, 14, and 17). These results show, for the first time, that exposure to UV-B can lead to cellular decline through active and regulated processes involving many genes also associated with natural senescence. PMID- 11432957 TI - Arabidopsis coactivator ALY-like proteins, DIP1 and DIP2, interact physically with the DNA-binding domain of the Zn-finger poly(ADP-ribose) polymerase. AB - Two novel homologous Arabidopsis proteins, DIP1 and DIP2, interact with the DNA binding domain of plant poly(ADP-ribose) polymerase (PARP) in the yeast two hybrid system and in vitro. Their homology to the transcriptional coactivator ALY suggests that plant PARP may play a role in the regulation of transcription. PMID- 11432958 TI - Identification of novel cyclin-dependent kinases interacting with the CKS1 protein of Arabidopsis. AB - The SUC1/CKS1 proteins interact with cyclin-dependent kinases (CDKs) and play an essential, but yet not entirely resolved, role in the regulation of the cell cycle. With the Arabidopsis thaliana CKS1At protein as bait in a two-hybrid screen, two novel Arabidopsis CDKs, Arath;CDKB1;2 and Arath;CDKB2;1, were isolated. A closely related homologue of Arath;CDKB2;1 was discovered in the databases and was nominated Arath;CDKB2;2. Transcript analysis of the five known Arath;CDKA and Arath;CDKB genes revealed that they all had the highest expression in flowers and cell suspensions. Differences in the expression patterns in roots, leaves and stems suggest unique roles for each CDK. PMID- 11432959 TI - Cultural aspects of cancer genetics: setting a research agenda. AB - BACKGROUND: Anecdotal evidence suggests that people from non-Anglo-Celtic backgrounds are under-represented at familial cancer clinics in the UK, the USA, and Australia. This article discusses cultural beliefs as a potential key barrier to access, reviews previous empirical research on cultural aspects of cancer genetics, draws implications from findings, and sets a research agenda on the inter-relationships between culture, cancer genetics, and kinship. METHODS: The CD-ROM databases MEDLINE, PsychLIT, CINAHL, and Sociological Abstracts were searched from 1980 onwards. RESULTS: Cultural aspects of cancer genetics is the focus of an emerging body of publications. Almost all studies assessed African American women with a family history of breast cancer and few studies included more diverse samples, such as Americans of Ashkenazi Jewish background or Hawaiian- and Japanese-Americans. Our analysis of published reports suggests several directions for future research. First, an increased focus on various Asian societies appears warranted. Research outside North America could explore the extent to which findings can be replicated in other multicultural settings. In addition, control group designs are likely to benefit from systematically assessing culture based beliefs and cultural identity in the "majority culture" group used for comparative purposes. CONCLUSION: More data on which to base the provision of culturally appropriate familial cancer clinic services to ethnically diverse societies are needed. Empirical data will assist with culturally appropriate categorisation of people from other cultures into risk groups based on their family histories and provide the basis for the development of culturally appropriate patient education strategies and materials. PMID- 11432960 TI - Genotype-phenotype correlation in hereditary multiple exostoses. AB - Hereditary multiple exostoses (HME) is a genetically heterogeneous autosomal dominant disorder characterised by the development of bony protuberances mainly located on the long bones. Three HME loci have been mapped to chromosomes 8q24 (EXT1), 11p11-13 (EXT2), and 19p (EXT3). The EXT1 and EXT2 genes encode glycosyltransferases involved in biosynthesis of heparan sulphate proteoglycans. Here we report on a clinical survey and mutation analysis of 42 HME French families and show that EXT1 and EXT2 accounted for more than 90% of HME cases in our series. Among them, 27/42 cases were accounted for by EXT1 (64%, four nonsense, 19 frameshift, three missense, and one splice site mutations) and 9/42 cases were accounted for by EXT2 (21%, four nonsense, two frameshift, two missense, and one splice site mutation). Overall, 31/36 mutations were expected to cause loss of protein function (86%). The most severe forms of the disease and malignant transformation of exostoses to chondrosarcomas were associated with EXT1 mutations. These findings provide the first genotype-phenotype correlation in HME and will, it is hoped, facilitate the clinical management of these patients. PMID- 11432961 TI - Segregation of a totally skewed pattern of X chromosome inactivation in four familial cases of Rett syndrome without MECP2 mutation: implications for the disease. AB - BACKGROUND: Rett syndrome is a neurodevelopmental disorder affecting only girls; 99.5% of Rett syndrome cases are sporadic, although several familial cases have been reported. Mutations in the MECP2 gene were identified in approximately 70 80% of sporadic Rett syndrome cases. METHODS: We have screened the MECP2 gene coding region for mutations in five familial cases of Rett syndrome and studied the patterns of X chromosome inactivation (XCI) in each girl. RESULTS: We found a mutation in MECP2 in only one family. In the four families without mutation in MECP2, we found that (1) all mothers exhibit a totally skewed pattern of XCI; (2) six out of eight affected girls also have a totally skewed pattern of XCI; and (3) it is the paternally inherited X chromosome which is active in the patients with a skewed pattern of XCI. Given that the skewing of XCI is inherited in our families, we genotyped the whole X chromosome using 32 polymorphic markers and we show that a locus potentially responsible for the skewed XCI in these families could be located on the short arm of the X chromosome. CONCLUSION: These data led us to propose a model for familial Rett syndrome transmission in which two traits are inherited, an X linked locus abnormally escaping X chromosome inactivation and the presence of a skewed XCI in carrier women. PMID- 11432962 TI - Disruption of one intra-chain disulphide bond in the carboxyl-terminal propeptide of the proalpha1(I) chain of type I procollagen permits slow assembly and secretion of overmodified, but stable procollagen trimers and results in mild osteogenesis imperfecta. AB - Type I procollagen is a heterotrimer comprised of two proalpha1(I) chains and one proalpha2(I) chain. Chain recognition, association, and alignment of proalpha chains into correct registration are thought to occur through interactions between the C-terminal propeptide domains of the three chains. The C-propeptide of each chain contains a series of cysteine residues (eight in proalpha1(I) and seven in proalpha2(I)), the last four of which form intra-chain disulphide bonds. The remaining cysteine residues participate in inter-chain stabilisation. Because these residues are conserved, they are thought to be important for folding and assembly of procollagen. We identified a mutation (3897C-->G) that substituted tryptophan for the cysteine at position 1299 in proalpha1(I) (C1299W, the first cysteine that participates in intra-chain bonds) and resulted in mild osteogenesis imperfecta. The patient was born with a fractured clavicle and four rib fractures. By 18 months of age he had had no other fractures and was on the 50th centile for length and weight. The proband's mother, maternal aunt, and grandfather had the same mutation and had few fractures, white sclerae, and discoloured teeth, but their heights were within the normal range. In the patient's cells the defective chains remained as monomers for over 80 minutes (about four times normal) and were overmodified. Some secreted procollagens were also overmodified but had normal thermal stability, consistent with delayed, but normal helix formation. This intra-chain bond may stabilise the C-propeptide and promote rapid chain association. Other regions of the C-propeptide thus play more prominent roles in chain registration and triple helix nucleation. PMID- 11432963 TI - Wild type Huntingtin reduces the cellular toxicity of mutant Huntingtin in mammalian cell models of Huntington's disease. AB - OBJECTIVES: Recent data suggest that wild type huntingtin can protect against apoptosis in the testis of mice expressing full length huntingtin transgenes with expanded CAG repeats. It is not clear if this protective effect was confined to particular cell types, or if wild type huntingtin exerted its protective effect in this model by simply reducing the formation of toxic proteolytic fragments from mutant huntingtin. METHODS: We cotransfected neuronal (SK-N-SH, human neuroblastoma) and non-neuronal (COS-7, monkey kidney) cell lines with HD exon 1 (containing either 21 or 72 CAG repeats) construct DNA and either full length wild type huntingtin or pFLAG (control vector). RESULTS: Full length wild type huntingtin significantly reduced cell death resulting from the mutant HD exon 1 fragments containing 72 CAG repeats in both cell lines. Wild type huntingtin did not significantly modulate cell death caused by transfection of HD exon 1 fragments containing 21 CAG repeats in either cell line. CONCLUSIONS: Our results suggest that wild type huntingtin can significantly reduce the cellular toxicity of mutant HD exon 1 fragments in both neuronal and non-neuronal cell lines. This suggests that wild type huntingtin can be protective in different cell types and that it can act against the toxicity caused by a mutant huntingtin fragment as well as against a full length transgene. PMID- 11432964 TI - A majority of fragile X males with methylated, full mutation alleles have significant levels of FMR1 messenger RNA. AB - FMR1 mRNA levels were determined in peripheral blood leucocytes for 48 fragile X males with methylated, full mutation alleles that are resistant to cleavage by methylation sensitive enzymes. Using quantitative (fluorescence) RT-PCR, we observed that more than half of these males produce FMR1 mRNA, with some mRNA levels approaching those found in normal subjects. In none of the samples analysed was there any evidence of premutation alleles. These results suggest that the assumed relationship between enzyme resistance and FMR1 gene silencing may not be generally valid. Despite the presence of FMR1 mRNA in some subjects, no FMRP production was detected by either immunocytochemistry or western blotting. The low/absent FMRP levels are probably a reflection of a post trancriptional effect such as a defect in translation. PMID- 11432965 TI - Distal arthrogryposis: a new type with distinct facial appearance and absent teeth. PMID- 11432966 TI - A Chinese adult onset type II citrullinaemia patient with 851del4/1638ins23 mutations in the SLC25A13 gene. PMID- 11432967 TI - Genetic analysis of the connexin-26 M34T variant. PMID- 11432968 TI - Gene expression analysis in N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mice model of Parkinson's disease using cDNA microarray: effect of R-apomorphine. AB - To establish the possible roles of oxidative stress, inflammatory processes and other unknown mechanisms in neurodegeneration, we investigated brain gene alterations in N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mice model of Parkinson's disease using Atlas mouse cDNA expression array membrane. The expression of 51 different genes involved in oxidative stress, inflammation, glutamate and neurotrophic factors pathways as well as in still undefined processes, such as cell cycle regulators and signal transduction molecules, was differentially affected by the treatment. The present study indicates the involvement of an additional cascade of events that might act in parallel to oxidative stress and inflammation to converge eventually into a common pathway leading to neurodegeneration. The attenuation of these gene changes by R apomorphine, an iron chelator-radical scavenger drug, supports our previous findings in vivo where R-apomorphine was neuroprotective. PMID- 11432969 TI - Elevated levels of group-III metabotropic glutamate receptors in the inferior colliculus of genetically epilepsy-prone rats following intracollicular administration of L-serine-O-phosphate. AB - The selective group-III metabotropic glutamate receptor agonist, L-serine-O phosphate (L-SOP), when injected bilaterally into the inferior colliculus of the sound sensitive genetically epilepsy-prone (GEP) rats produces a short proconvulsant excitation followed by a long phase of protection against sound induced seizures lasting for 2-4 days. We have studied this prolonged suppression of audiogenic seizures using pharmacological and molecular biological approaches including semiquantitative RT-PCR and western blotting. The intracerebroventricular injection of the protein synthesis inhibitor cycloheximide (120 microg) 30 min beforehand significantly reduces the proconvulsant seizure activity and the prolonged anticonvulsant effect of intracollicular L-SOP (500 nmol/side). The sensitive semiquantitative RT-PCR revealed a significant up-regulation in mGlu(4) and mGlu(7) mRNA levels in the inferior colliculus at 2 days (maximum suppression of audiogenic seizures) after intracollicular L-SOP injection compared with the non-injected, 2-day post vehicle treated and 7-day (return to expressing audiogenic seizures) post-drug or vehicle-treated groups. No significant changes were observed in mGlu(6) or mGlu(8) mRNA expression levels in drug-treated compared with control groups. Examination of mGlu(4a) and mGlu(7a) protein levels using western blotting showed a significant increase in mGlu(7a) but no significant change in mGlu(4a) protein levels 2 days after L-SOP treatment compared with the control groups (non injected and 2-day vehicle-injected group). These results suggest that up regulation of mGlu(7) receptors is involved in the prolonged anticonvulsant effect of L-SOP against sound-induced seizures in GEP rats. The potential use of mGlu(7) agonists as novel anti-epileptic agents merits investigation. PMID- 11432970 TI - Mimicking the nicotinic receptor binding site by a single chain Fv selected by competitive panning from a synthetic phage library. AB - We have developed a novel competitive method to select from a phage display library a single chain Fv which is able to mimic the alpha-bungarotoxin binding site of the muscle nicotinic receptor. The single chain Fv was selected from a large synthetic library using alpha-bungarotoxin-coated magnetic beads. Toxin bound phages were then eluted by competition with affinity purified nicotinic receptor. Recognition of the toxin by the anti-alpha-bungarotoxin single chain Fv was very similar to that of the receptor, such as indicated by the epitope mapping of alpha-bungarotoxin through overlapping synthetic peptides. Moreover, several positively charged residues located in the toxin second loop and in the C terminal region were found to be critical, to a similar extent, for toxin recognition by the single chain Fv and the receptor. However, although the anti alpha-bungarotoxin single chain Fv seems to mimic the toxin binding site of the nicotinic receptor, it does not bind other nicotinic agonists or antagonists. Our results suggest that competitive selection of anti-ligand antibody phages can allow the production of receptor-mimicking molecules directly and exclusively targeted at one specific ligand. Since physiologically and pharmacologically different ligands can produce opposite effects on receptor functions, such selective ligand decoys can have important therapeutic applications. PMID- 11432971 TI - Effect of proteasome inhibition on cellular oxidative damage, antioxidant defences and nitric oxide production. AB - The ubiquitin/proteasome pathway plays an essential role in protein turnover in vivo, and contributes to removal of oxidatively damaged proteins. We examined the effects of proteasome inhibition on viability, oxidative damage and antioxidant defences in NT-2 and SK-N-MC cell lines. The selective proteasome inhibitor, lactacystin (1 microM) caused little loss of viability, but led to significant increases in levels of oxidative protein damage (measured as protein carbonyls), ubiquitinated proteins, lipid peroxidation and 3-nitrotyrosine, a biomarker of the attack of reactive nitrogen species (such as peroxynitrite, ONOO(-)) upon proteins. Higher levels (25 microM) of lactacystin did not further increase the levels of carbonyls, lipid peroxidation, 3-nitrotyrosine, or ubiquitinated proteins, but produced increases in the levels of 8-hydroxyguanine (a biomarker of oxidative DNA damage) and falls in levels of GSH. Lactacystin (25 microM) caused loss of viability, apparently by apoptosis, and also increased production of nitric oxide (NO.) (measured as levels of NO2- plus NO3-) by the cells; this was inhibited by N-nitro-L-arginine methyl ester (L-NAME), which also decreased cell death induced by 25 microM lactacystin and decreased levels of 3 nitrotyrosine. The NO. production appeared to involve nNOS; iNOS or eNOS were not detectable in either cell type. Another proteasome inhibitor, epoxomicin, had similar effects. PMID- 11432972 TI - Parkin is associated with cellular vesicles. AB - We recently identified a novel gene, parkin, as a pathogenic gene for autosomal recessive juvenile parkinsonism. Parkin encodes a 52-kDa protein with a ubiquitin like domain and two RING-finger motifs. To provide a insight into the function of parkin, we have examined its intracellular distribution in cultured cells. We found that parkin was localized in the trans-Golgi network and the secretory vesicles in U-373MG or SH-SY5Y cells by immunocytochemical analyses. In the subsequent subcellular fractionation studies of rat brain, we showed that parkin was copurified with the synaptic vesicles (SVs) when we used low ionic conditions throughout the procedure. An immunoelectromicroscopic analysis indicated that parkin was present on the SV membrane. Parkin was readily released from SVs into the soluble phase by increasing ionic strength at neutral pH, but not by a non ionic detergent. To elucidate its responsible region for membrane association, we transfected with green fluorescent protein-tagged deletion mutants of parkin into COS-1 cells followed by subcellular fractionation. We demonstrated the ability of parkin to bind to the membranes through a broad region except for the ubiquitin like domain. The significance of SV localization of parkin is discussed. PMID- 11432973 TI - Morphine-induced in vivo release of spinal cholecystokinin is mediated by delta opioid receptors--effect of peripheral axotomy. AB - Morphine and other opioid agonists induce spinal in vivo release of cholecystokinin (CCK), a neuropeptide with anti-opioid properties. However, so far the opioid receptor subtype responsible for this effect has not been determined. In the present in vivo microdialysis study, the morphine-induced release of cholecystokinin-like immunoreactivity (CCK-LI) in the dorsal horn was completely blocked by the delta-opioid antagonist naltrindole (10 microM in the perfusion fluid). Neither the mu-opioid receptor antagonist D-Phe-Cys-Tyr-D-Trp Orn-Thr-Pen-Thr amide (CTOP; 10 microM in the perfusion fluid), nor the kappa opioid receptor antagonist nor-binaltorphimine (nor-BNI); 10 microM in the perfusion fluid) had any significant effect in this respect. In addition, systemic administration of the delta-opioid receptor agonist BW373U86 (1 mg/kg, s.c.) and spinal administration of the delta(2)-opioid receptor agonist, Tyr-D Ala-Phe-Glu-Val-Val-Gly amide ([D-Ala(2)] deltorphin II) (1 microM in the perfusion fluid) induced a significant increase of the CCK-LI level. The effect of BW373U86 on spinal CCK-LI release was completely blocked by spinal administration of naltrindole. The mu-opioid receptor agonist [D-ala(2)-N-Me Phe(4)-Gly(5)-ol]-enkephalin (DAMGO) (1 microM in the perfusion fluid or 1 mg/kg, s.c.) failed to alter the CCK-LI level. Peripheral nerve lesions have previously been shown to down-regulate mu- and delta-opioid receptors in the dorsal horn, to increase the gene-expression of CCK and CCK-receptor mRNA in dorsal root ganglion neurons and to alter the potassium-induced spinal CCK-LI release. After complete sciatic nerve transection, administration of the two selective delta-opioid receptor agonists induced a significant release of CCK-LI, which was comparable to controls. In contrast, neither systemic nor spinal administration of morphine and DAMGO altered the spinal CCK-LI release in axotomized animals. The present data indicate that the delta-opioid receptor mediates morphine-induced CCK-LI release in the spinal cord. PMID- 11432974 TI - The specificity of monoclonal antibody A2B5 to c-series gangliosides. AB - To examine the specificity of monoclonal antibody A2B5, four A2B5-reactive gangliosides (designated as G-1, G-2, G-3 and G-4) were purified from bonito fish brain. Ganglioside-1, -2, and -3 migrated above GD1b, below GQ1b, and far below GQ1b on thin-layer chromatography. Ganglioside-4 had the slowest chromatographic mobility and migrated below G-3. The structures of these gangliosides were characterized by overlay analysis with glycolipid-specific ligands, product analysis after sialidase or mild acid treatment, and electrospray ionization-mass spectrometry (ESI-MS). Accordingly, G-1, G-2 and G-3 were identified to be GT3, GQ1c and GP1c, respectively. The ganglioside G-4 was shown to have the following structure: NeuAc-NeuAc-NeuAc-Galbeta1-3Gal NAcbeta1-4(NeuAc-NeuAc-NeuAcalpha2 3)Galbeta1-4Glcbeta1-1'Cer. The antibody A2B5 reacted with these c-series gangliosides, but not with GD3 and other gangliosides and neutral glycosphingolipids. The antigenic epitope for A2B5 was assumed to include the trisialosyl residue connected to the inner galactose of the hemato- or ganglio type oligosaccharide structure of gangliosides. Phylogenetic analysis of brain gangliosides using the A2B5 preparation demonstrated that c-series gangliosides are enriched in lower animals, especially bony fish of different species. The monoclonal antibody A2B5 would be a useful tool for examining the distribution and function of c-series gangliosides. PMID- 11432975 TI - Molecular cloning and characterization of neural activity-related RING finger protein (NARF): a new member of the RBCC family is a candidate for the partner of myosin V. AB - Activity-dependent synaptic plasticity has been thought to be a cellular basis of memory and learning. The late phase of long-term potentiation (L-LTP), distinct from the early phase, lasts for up to 6 h and requires de novo synthesis of mRNA and protein. Many LTP-related genes are enhanced in the hippocampus during pentyrenetetrazol (PTZ)- and kainate (KA)-mediated neural activation. In this study, mice were administered intraperitoneal injections of PTZ 10 times, once every 48 h, and showed an increase in seizure indexes. Genes related to plasticity were efficiently induced in the mouse hippocampus. We used a PCR-based cDNA subtraction method to isolate genes that are expressed in the hippocampus of repeatedly PTZ-treated mice. One of these genes, neural activity-related RING finger protein (NARF), encodes a new protein containing a RING finger, B-box zinc finger, coiled-coil (RBCC domain) and beta-propeller (NHL) domain, and is predominantly expressed in the brain, especially in the hippocampus. In addition, KA up-regulated the expression of NARF mRNA in the hippocampus. This increase correlated with the activity of the NMDA receptor. By analysis using GFP-fused NARF, the protein was found to localize in the cytoplasm. Enhanced green fluorescent protein-fused NARF was also localized in the neurites and growth cones in neuronal differentiated P19 cells. The C-terminal beta-propeller domain of NARF interacts with myosin V, which is one of the most abundant myosin isoforms in neurons. The NARF protein increases in hippocampal and cerebellar neurons after PTZ-induced seizure. These observations indicated that NARF expression is enhanced by seizure-related neural activities, and NARF may contribute to the alteration of neural cellular mechanisms along with myosin V. PMID- 11432976 TI - Expression and role of phosphodiesterase 6 in the chicken pineal gland. AB - The chicken pineal gland is directly photosensitive, with light causing an inhibition of melatonin synthesis. A possible role of phosphodiesterase 6 (PDE6, the primary effector of retinal phototransduction) in mediating this response was investigated. RT-PCR, DNA sequencing and northern blots revealed the presence of RNA encoding both catalytic and regulatory subunits of PDE6 in the chicken pineal gland. Both rod and cone forms of PDE6 subunits mRNA were detected. The concentration of the transcripts encoding PDE6 catalytic subunits peaked at night. Western blot analysis of chicken pineal proteins with an antibody directed against the catalytic subunits of bovine rod PDE6 identified a single immunoreactive protein of 97 kDa. Anion exchange chromatography of chicken pineal soluble proteins revealed a peak of PDE6 activity that accounted for about 30% of cyclic GMP-hydrolysis. In cultured chick pineal glands, arylalkylamine N acetyltransferase (AA-NAT), the rate-limiting enzyme of melatonin synthesis, was protected from inhibition by light when selective PDE5/6 inhibitors (zaprinast, DMPPO) were added to the culture medium. PDE5/6 inhibitors did not affect AA-NAT activity in the dark. In contrast, a general PDE inhibitor (IBMX) increased AA NAT in a light-independent manner. Together, the data indicate that rod and cone forms of PDE6 are expressed in chick pineal cells and that this enzyme plays a role in the inhibition of melatonin synthesis by light. PMID- 11432977 TI - Different action on dopamine catabolic pathways of two endogenous 1,2,3,4 tetrahydroisoquinolines with similar antidopaminergic properties. AB - The effect of single and multiple 1-methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ) and 1-benzyl-1,2,3,4-tetrahydroisoquinoline (1BnTIQ) administration on concentrations of dopamine and its metabolites: homovanillic acid (HVA) and 3,4 dihydroxyphenylacetic acid (DOPAC) and 3-methoxytyramine (3MT) in three brain areas was studied HPLC with electrochemical detection in Wistar rats. The rate of dopamine catabolism in the striatum along the N-oxidative and O-methylation pathways was assessed by calculation of the ratio of appropriate metabolites to dopamine concentration. In addition, the spontaneous and apomorphine-stimulated locomotor activity, and muscle rigidity was studied after acute administration of 1MeTIQ and 1BnTIQ. We have found that 1MeTIQ did not change the level of dopamine and HVA in all investigated structures both after a single and chronic administration. However, the levels of intermediary dopamine metabolites, DOPAC and 3MT, were distinctly affected. The level of DOPAC was strongly depressed (by 60-70%) while the level of extraneuronal matabolite 3MT was significantly elevated (by 170-200%). In contrast to 1MeTIQ, 1BnTIQ depressed the level of dopamine (by approximately 60%) and increased the level of total metabolite, HVA, (by 40%) especially in the striatum, but the levels of DOPAC and 3MT remained unchanged. The paper has shown that 1MeTIQ and 1BnTIQ produced different effects on dopamine catabolism. Potential neuroprotective compound 1MeTIQ did not change the rate of total dopamine catabolism, it strongly inhibited the monoamine oxidase (MAO)-dependent catabolic pathway and significantly activated the catechol-O-methyltransferase (COMT)-dependent O-methylation. In contrast 1BnTIQ, a compound with potential neurotoxic activity, produced the significant increase of the rate of dopamine metabolism with strong activation of the oxidative MAO dependent catabolic pathway. Interestingly, both compounds produced similar antidopaminergic functional effects: antagonism of apomorphine hyperactivity and induction of muscle rigidity. The results may explain the biochemical basis of the neuroprotective and of the neurotoxic properties endogenous brain tetrahydroisoquinoline derivatives. PMID- 11432978 TI - C-terminal fragment of amyloid precursor protein induces astrocytosis. AB - One of the pathophysiological features of Alzheimer's disease is astrocytosis around senile plaques. Reactive astrocytes may produce proinflammatory mediators, nitric oxide, and subsequent reactive oxygen intermediates such as peroxynitrites. In the present study, we investigated the possible role of the C terminal fragment of amyloid precursor protein (CT-APP), which is another constituent of amyloid senile plaque and an abnormal product of APP metabolism, as an inducer of astrocytosis. We report that 100 nM recombinant C-terminal 105 amino acid fragment (CT105) of APP induced astrocytosis morphologically and immunologically. CT105 exposure resulted in activation of mitogen-activated protein kinase (MAPK) pathways as well as transcription factor NF-kappaB. Pretreatment with PD098059 and/or SB203580 decreased nitric oxide (NO) production and nuclear factor-kappa B (NF-kappaB) activation. But inhibitors of NF-kappaB activation did not affect MAPKs activation whereas they abolished NO production and attenuated astrocytosis. Furthermore, conditioned media derived from CT105 treated astrocytes enhanced neurotoxicity and pretreatment with NO and peroxynitrite scavengers attenuated its toxicity. These suggest that CT-APP may participate in Alzheimer's pathogenesis through MAPKs- and NF-kappaB-dependent astrocytosis and iNOS induction. PMID- 11432979 TI - Induction of NADPH cytochrome P450 reductase by the Alzheimer beta-protein. Amyloid as a "foreign body". AB - A large body of data suggests that the Alzheimer's amyloid peptide (Abeta) causes degeneration and death of neurons by mechanisms that involve reactive oxygen species. The pathways involved in Abeta-mediated oxidative injury are only partially understood. We theorized that abnormal microaggregates and/or pathological conformations of Abeta peptides may behave as xenobiotics and trigger the induction of NADPH cytochrome P450 reductase (CP450r), an enzyme which, if induced by non-physiological substrates (such as xenobiotics like drugs or other 'foreign molecules'), is known to cause oxidative stress. In order to test this hypothesis, i.e. that Abeta can increase the expression of CP450r, SK-N SH human neuroblastoma cells were exposed to Abeta25-35 and Abeta1-42 and then examined for induction of this enzyme in immunoblots, using specific antibodies. Following exposure to Abeta peptides, neuroblastoma cells showed a clear-cut induction of CP450r. To determine whether this mechanism is operational in vivo, we investigated the expression of CP450r in a transgenic mouse model of Alzheimer's disease (AD) and in brains from patients afflicted with AD, using an immunocytochemical approach. Tissue sections from brains of transgenic mice exhibited strong immunoreactivity for CP450r, surrounding amyloid deposits. The pattern of expression of CP450r was similar to that exhibited by neuritic and oxidative stress markers. Sections from non-transgenic mice showed no detectable immunoreactivity. Immunostaining of sections from four brains with neuropathologically confirmed AD showed a pattern of abnormality different from transgenic mice that was characterized by abnormal immunoreactivity for CP450r within the cytoplasm of cortical neurons. No labeling was seen in sections from aged-matched control brains. The data showed that CP450r is induced by Alzheimer amyloid peptide and that such a response must be considered as one possible mechanism whereby Abeta causes oxidative stress. PMID- 11432980 TI - A 27-bp region of the inducible nitric oxide synthase promoter regulates expression in glial cells. AB - The expression of inducible nitric oxide synthase (NOS2) in glial cells is inhibited by neurotransmitters such as norepinephrine (NE) which elevate cAMP levels. We examined the molecular basis for this effect using a 2.2-kb fragment of the rat NOS2 promoter transfected into rat C6 glioma cells. Promoter activation (up to six-fold) by lipopolysaccharide (LPS) and interferon-gamma (IFNgamma) was reduced by NE, which alone had no effect. However, a promoter construct extending to bp -130 and containing the proximal nuclear factor-kappa B (NF-kappaB) binding site was minimally activated by LPS and cytokines, but activated up to three-fold by NE. Deletion analysis identified a 27-bp region (bp -187 to -160) as critical for mediating this suppressive effect. This region also enhanced promoter activation by LPS and cytokines, and prevented activation by NE alone. Gel shift analysis revealed constitutive binding to this region, and induction by NE of additional complexes which could be blocked by an antibody against CREB. NE also increased levels of the IkappaBalpha protein which could contribute to its suppressive effects. These results identify a critical role for this 27-bp region in regulation of NOS2 promoter activation and suppression by cAMP. PMID- 11432981 TI - Role of DNA-dependent protein kinase in neuronal survival. AB - DNA-dependent protein kinase (DNA-PK) is a DNA repair enzyme composed of a DNA binding component called Ku70/80 and a catalytic subunit called DNA-PKcs. Many investigators have utilized DNA-PKcs-deficient cells and cell lines derived from severe combined immunodeficiency (scid) mice to study DNA repair and apoptosis. However, little is known about the CNS of these mice. This study was carried out using primary neuronal cultures derived from the cerebral hemispheres of new-born wild-type and scid mice to investigate the effects of loss of DNA-PK function on neuronal maturation and survival. Purified neuronal cultures developed comparably in terms of neurite formation and expression of neuronal markers, but scid cultures showed a significant increase in the percentage of dying cells. Furthermore, when apoptosis was induced by staurosporine, scid neurons died more rapidly and in higher numbers. Apoptotic scid neurons exhibited nuclear condensation, DNA fragmentation and caspase-3 activation, but treatment with the general caspase inhibitor, N-benzyloxycarbonyl-Val-Ala-Asp-(O-methyl) fluoromethyl ketone did not prevent staurosporine-induced apoptosis. We conclude that a DNA-PK deficiency in cultured scid neurons may cause an accumulation of DNA damage and increased susceptibility to caspase-independent forms of programmed cell death. PMID- 11432982 TI - Down-regulation of occludin expression in astrocytes by tumour necrosis factor (TNF) is mediated via TNF type-1 receptor and nuclear factor-kappaB activation. AB - Tight junctions form the diffusion barrier of brain microcapillary endothelial cells and support cell polarity. Also astrocytes express tight junction components such as occludin, claudin-1, ZO-1 and ZO-2, but do not establish a permeability barrier. However, little is known about the function and regulation of these molecules in astrocytes. We studied the impact of tumour necrosis factor (TNF) on occludin and ZO-1 expression in astrocytes. TNF decreased occludin, but not ZO-1 expression. In brain microcapillary endothelial cells, as well as in epithelial cells, occludin expression was not influenced by TNF. Removal of TNF from astrocytes restored the basal level of occludin. Down-regulation was inhibited by caffeic acid phenethyl ester, a specific inhibitor of nuclear factor kappaB (NF-kappaB) activation. Exposure of astrocytes isolated from mice deficient in either TNF type-1 receptor (TNFR1), TNF type-2 receptor (TNFR2), or both, respectively, revealed that down-regulation was mediated entirely by TNFR1. ZO-1, which can interact with occludin, was found to co-precipitate connexin43, but not occludin. These findings demonstrate that TNF selectively down-regulates occludin in astrocytes, but not in cells forming established tight junctions, through TNFR1 and suggest that NF-kappaB is involved as a negative regulator. PMID- 11432983 TI - Survival promotion of mesencephalic dopaminergic neurons by depolarizing concentrations of K+ requires concurrent inactivation of NMDA or AMPA/kainate receptors. AB - The death of dopaminergic neurons that occurs spontaneously in mesencephalic cultures was prevented by depolarizing concentrations of K+ (20-50 mM). However, unlike that observed previously in other neuronal populations of the PNS or CNS, promotion of survival required concurrent blockade of either NMDA or alpha-amino 3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)/kainate receptors by the specific antagonists, MK-801 and GYKI-52466, respectively. Rescued neurons appeared to be healthy and functional because the same treatment also dramatically enhanced their capacity to accumulate dopamine. The effects on survival and uptake were rather specific to dopaminergic neurons, rapidly reversible and still observed when treatment was delayed after plating. Glutamate release increased substantially in the presence of elevated concentrations of K+, and chronic treatment with glutamate induced a loss of dopaminergic neurons that was prevented by MK-801 or GYKI-52466 suggesting that an excitotoxic process interfered with survival when only the depolarizing treatment was applied. The effects of the depolarizing stimulus in the presence of MK-801 were mimicked by BAY K-8644 and abolished by nifedipine, suggesting that neuroprotection resulted from Ca(2+) influx through L-type calcium channels. Measurement of intracellular calcium revealed that MK-801 or GYKI-52466 were required to maintain Ca(2+) levels within a trophic range, thus preventing K+-induced excitotoxic stress and Ca(2+) overload. Altogether, our results suggest that dopaminergic neurons may require a finely tuned interplay between glutamatergic receptors and calcium channels for their development and maturation. PMID- 11432984 TI - Glutamate(NMDA) receptor NR1 subunit mRNA expression in Alzheimer's disease. AB - We analyzed the expression profile of two NMDAR1 mRNA isoform subsets, NR1(0XX) and NR1(1XX), in discrete regions of human cerebral cortex. The subsets are characterized by the absence or presence of a 21-amino acid N-terminal cassette. Reverse transcription polymerase chain reaction for NR1 isoforms was performed on total RNA preparations from spared and susceptible regions from 10 pathologically confirmed Alzheimer's disease (AD) cases and 10 matched controls. Primers spanning the splice insert yielded two bands, 342 bp (NR1(0XX)) and 405 bp (NR1(1XX)), on agarose gel electrophoresis. The bands were visualized with ethidium and quantified by densitometry. NR1(1XX) transcript expression was calculated as a proportion of the NR1(1XX) + NR1(0XX) total. Values were significantly lower in AD cases than in controls in mid-cingulate cortex, p < 0.01, superior temporal cortex, p < 0.01 and hippocampus, p approximately 0.05. Cortical proportionate NR1(1XX) transcript expression was invariant over the range of ages and areas of controls tested, at approximately 50%. This was also true for AD motor and occipital cortex. Proportionate NR1(1XX) expression in AD cingulate and temporal cortex was lower at younger ages and increased with age: this regression was significantly different from that in the homotropic areas of controls. Variations in NR1 N-terminal cassette expression may underlie the local vulnerability to excitotoxic damage of some areas in the AD brain. Alternatively, changes in NR1 mRNA expression may arise as a consequence of the AD disease process. PMID- 11432985 TI - Functional striatal hypodopaminergic activity in mice lacking adenosine A(2A) receptors. AB - Adenosine and caffeine modulate locomotor activity and striatal gene expression, partially through the activation and blockade of striatal A(2A) receptors, respectively. The elucidation of the roles of these receptors benefits from the construction of A(2A) receptor-deficient mice (A(2A)-R(-/-)). These mice presented alterations in locomotor behaviour and striatal expression of genes studied so far, which are unexpected regarding the specific expression of A(2A) receptor by striatopallidal neurones. To clarify the functions of A(2A) receptors in the striatum and to identify the mechanisms leading to these unexpected modifications, we studied the basal expression of immediate early and constitutive genes as well as dopamine and glutamate neurotransmission in the striatum. Basal zif268 and arc mRNAs expression was reduced in mutant mice by 60 80%, not only in the striatum but also widespread in the cerebral cortex and hippocampus. Striatal expression of substance P and enkephalin mRNAs was reduced by about 50% and 30%, respectively, whereas the expression of GAD67 and GAD65 mRNAs was slightly increased and unaltered, respectively. In vivo microdialysis in the striatum revealed a 45% decrease in the extracellular dopamine concentration and three-fold increase in extracellular glutamate concentration. This was associated with an up-regulation of D(1) and D(2) dopamine receptors expression but not with changes in ionotropic glutamate receptors. The levels of tyrosine hydroxylase and of striatal and cortical glial glutamate transporters as well as adenosine A(1) receptors expression were indistinguishable between A(2A) R(-/-) and wild-type mice. Altogether these results pointed out that the lack of A(2A) receptors leads to a functional hypodopaminergic state and demonstrated that A(2A) receptors are necessary to maintain a basal level in immediate early and constitutive genes expression in the striatum and cerebral cortex, possibly via their control of dopamine pathways. PMID- 11432986 TI - Lithium inhibits caspase 3 activation and dephosphorylation of PKB and GSK3 induced by K+ deprivation in cerebellar granule cells. AB - Lithium protects cerebellar granule cells from apoptosis induced by low potassium, and also from other apoptotic stimuli. However, the precise mechanism by which this occurs is not understood. When cerebellar granule cells were switched to low potassium medium, the activation of caspase 3 was detected within 6 h, suggesting a role of caspase 3 in mediating apoptosis under conditions of low potassium. In the same conditions, lithium (5 mM) inhibited the activation of caspase 3 induced by low potassium. As lithium did not inhibit caspase 3 activity in vitro, these results suggest that this ion inhibits an upstream component that is required for caspase 3 activation. Lithium is known to inhibit a kinase termed glycogen sythase kinase 3 (GSK3), which is implicated in the survival pathway of phosphatidylinositol 3-kinase/protein kinase B (PI3K/PKB). Here we demonstrate that low potassium in the absence of lithium induces the dephosphorylation, and therefore the activation, of GSK3. However, when lithium was present, GSK3 remained phosphorylated at the same level as observed under conditions of high potassium. Low potassium induced the dephosphorylation and inactivation of PKB, whereas when lithium was present PKB was not dephosphorylated. Our results allow us to propose a new hypothesis about the action mechanism of lithium, this ion could inhibit a serine-threonine phosphatase induced by potassium deprivation. PMID- 11432987 TI - Functional characterization of the C-terminus of the human ether-a-go-go-related gene K(+) channel (HERG). AB - 1. In the present study the functional role of the C-terminus of the human ether a-go-go-related gene K(+) channel HERG was investigated using a series of C terminal deletion constructs expressed in Xenopus oocytes. 2. Constructs with deletions of 311 or more amino acid residues failed to form functional channels. Truncation by 215 amino acid residues or fewer had no discernable effects on channel activity. Truncation by 236 or 278 amino acid residues accelerated deactivation, and caused a faster recovery from inactivation. 3. In high extracellular K(+), channel deactivation of HERG results from the binding of the N-terminus to a site within the pore. This slows channel deactivation by a knock off mechanism. Here, it was shown that C-terminal deletions also abolished this effect of high extracellular K(+). Mutants containing deletions in both the N- and C-termini deactivated with rates similar to those observed in individual deletion mutants. 4. In contrast, experiments with double-deletion constructs showed additive effects of the N- and C-termini on the voltage dependence of activation, and on the kinetics of inactivation and recovery from inactivation. The reduction of inactivation in these mutants contributed to an increase in peak current amplitude. 5. These results indicate that residues within the C-terminus of HERG play a role in channel expression as well as in most aspects of channel gating. The regulation of channel deactivation is likely to be mediated by an interaction with the N-terminus, but the regulation of the voltage dependence of activation, and of rate processes associated with inactivation, does not require the N-terminus. PMID- 11432988 TI - Properties of single M-type KCNQ2/KCNQ3 potassium channels expressed in mammalian cells. AB - 1. The single channel properties of KCNQ2/KCNQ3 channels underlying neuronal voltage-dependent M-type potassium currents were studied in cell-attached patches from transfected Chinese hamster ovary (CHO) cells. Macroscopic currents produced by homo- and heteromeric KCNQ2/KCNQ3 channels were measured using the perforated patch whole-cell technique. 2. Compared with heteromeric KCNQ2 + KCNQ3 channels, homomeric KCNQ2 channels had lower slope conductance (9.0 +/- 0.3 and 5.8 +/- 0.3 pS, respectively) and open probability at 0 mV (0.30 +/- 0.07 and 0.15 +/- 0.03, respectively), consistent with their 3.8-fold smaller macroscopic currents. By contrast, homomeric KCNQ3 channels had the same slope conductance (9.0 +/- 1.1 pS) as KCNQ2 + KCNQ3 channels, and higher open probability (0.59 +/- 0.11), inconsistent with their 12.7-fold smaller macroscopic currents. Thus, KCNQ2 and KCNQ3 subunits may play different roles in the expression of M-type currents, with KCNQ2 ensuring surface expression of underlying channels and KCNQ3 modifying their function. 3. Both in homo- and heteromeric KCNQ2/KCNQ3 channels the shut time distributions were fitted with three, and the open time distributions with two, exponential components. By measuring these and other parameters (e.g. conductance and open probability) KCNQ2/ KCNQ3 channels can be shown to resemble previously characterised neuronal M-type channels. PMID- 11432989 TI - Functional evidence of distinct ATP activation sites at the human P2X(7) receptor. AB - 1. The effect of the agonist ATP on whole cell currents of Xenopus oocytes expressing either the wild-type human P2X(7) receptor (hP2X(7)), an N-terminally hexahistidyl-tagged hP2X(7) receptor (His-hP2X(7)), or a truncated His-hP2X(7) receptor (His-hP2X(7)DeltaC) lacking the C-terminal 156 amino acids was investigated using the two-microelectrode voltage clamp technique. 2. The activation time course of the wild-type hP2X(7) receptor can be described as the sum of an exponentially growing and an additional almost linearly activating current component. 3. The amplitude of the exponentially activating current component of the wild-type hP2X(7) receptor displayed a biphasic dependence on the agonist concentration, which could be best approximated by a model of two equal high-sensitivity and two equal low-sensitivity non-cooperative activation sites with apparent dissociation constants of about 4 and 200 microM free ATP(4 ), respectively. 4. The linearly activating current was monophasically dependent on the agonist concentration with an apparent dissociation constant of about 200 microM. 5. The contribution of the low-sensitivity sites to current kinetics was reduced or almost abolished in oocytes expressing His-hP2X(7) or His hP2X(7)DeltaC. 6. Our data indicate that the hP2X(7) receptor possesses at least two types of activation sites, which differ in ATP(4-) sensitivity by a factor of 50. The degree of occupation of these two sites influences both activation and deactivation kinetics. Both N- and C-terminal domains appear to be important determinants of the current elicited by activation of the sites with low ATP sensitivity, but not for that mediated by the highly ATP-sensitive sites. PMID- 11432990 TI - A comparison of the effects of ATP and tetracaine on spontaneous Ca(2+) release from rat permeabilised cardiac myocytes. AB - 1. Fluo-3 fluorescence measurements were made in isolated beta-escin permeablised rat cardiac myocytes using confocal microscopy. Perfusion of a mock intracellular solution containing 0.22-0.23 microM Ca(2+) and 5 mM ATP elicited regular waves of Ca(2+) (approximately every 5 s) due to spontaneous release of Ca(2+) from the sarcoplasmic reticulum (SR). 2. An approximately linear relationship was noted between Ca(2+) wave velocity (v) and amplitude (sigma). Under the control conditions the ratio of velocity to amplitude (v/sigma) varied little and was 99.8 +/- 2.5 m s(-1) microM(-1) (n = 78). 3. Reduction of [ATP] in the bathing solution to 0.5 and 0.2 mM ATP progressively decreased Ca(2+) wave frequency and propagation velocity while increasing the amplitude. The changes in Ca(2+) wave characteristics in 0.5 mM ATP were similar to those observed during perfusion with 50 microM tetracaine. In 0.2 mM ATP the decline of [Ca(2+)] during a Ca(2+) wave was slowed suggesting a lowered rate of Ca(2+) re-uptake by the SR Ca(2+) pump. 4. Reduction of [ATP] to 0.1 mM abolished Ca(2+) waves after 15-20 s. Returning the [ATP] to 5 mM caused a burst of high frequency and large amplitude waves. Mean velocity of the first wave on returning to 5 mM ATP was larger than normal but the v/sigma value was 32 +/- 6 % of control (n = 6). In the similar burst on removal of 100 microM tetracaine v/sigma was higher than control (166 +/ 9 %, n = 6). 5. Rapid application of caffeine (10 mM) was used to assess the SR Ca(2+) content. This showed that SR Ca(2+) increased as [ATP] was reduced or [tetracaine] was increased. The highest SR Ca(2+) content was observed after perfusion with 0.1 mM ATP, which was 245 +/- 15 % of control values. 6. Returning [ATP] from 0.1 mM to 5 mM caused a burst of high frequency, large amplitude Ca(2+) waves. But recovery after incubation with 300 microM tetracaine resulted in SR Ca(2+) release with no coherent wave pattern. The reason for this discrepancy is discussed. PMID- 11432991 TI - K(+)-dependent gating of K(ir)1.1 channels is linked to pH gating through a conformational change in the pore. AB - 1. We have used giant patch-clamp recording to investigate the interaction between pH gating and K(+)-dependent gating in rat K(ir)1.1 (ROMK) channels heterologously expressed in Xenopus oocytes. 2. Gating by intracellular protons (pH gating) and extracellular K(+) ions (K(+)-dependent gating) is a hallmark of K(ir)1.1 channels that mediate K(+) secretion and control NaCl reabsorption in the kidney. pH gating is driven by protonation of an intracellular lysine residue (K80 in K(ir)1.1). K(+)-dependent gating occurs upon withdrawal of K(+) ions from the extracellular side of the channel. Both gating mechanisms are thought to interact allosterically. 3. K(+)-dependent gating was shown to be strictly coupled to pH gating; it only occurred when channels were in the pH-inactivated closed state, but not in the open state. Moreover, K(+)-dependent gating was absent in the non-pH-gated mutant K(ir)1.1(K80 M). 4. Channels inactivated by K(+)-dependent gating were reactivated upon addition of permeant ions to the extracellular side of the membrane, while impermeant ions failed to induce channel reactivation. Moreover, mutagenesis identified two residues in the P helix (L136 and V140 in K(ir)1.1) that are crucial for K(+)-dependent gating. Replacement of these residues with the ones present in the non-K(+)-gated K(ir)2.1 abolished K(+)-dependent gating of K(ir)1.1 channels without affecting pH gating. 5. The results indicate that pH gating and K(+)-dependent gating are coupled to each other via structural rearrangements in the inner pore involving the P-helix. PMID- 11432992 TI - Coupling of a P2Z-like purinoceptor to a fatty acid-activated K(+) channel in toad gastric smooth muscle cells. AB - 1. Extracellular application of ATP generates two whole-cell currents in toad gastric smooth muscle cells: an immediate inward non-selective cation current (due to the activation of a P2X or P2Z-like receptor) and a slowly developing outward K(+) current. The inward non-selective cation current depends on the continuous presence of ATP while the outward K(+) current can last for minutes after ATP application ceases. 2. In cell-attached patches, application of ATP to the extra-patch membrane can activate K(+) channels in the patch indicating that a diffusible cellular messenger may be involved. The characteristics of these K(+) channels are similar to those of a previously described fatty acid-activated K(+) channel that is also a stretch-activated channel. 3. This whole-cell K(+) current can be induced by ATP in the absence of extracellular Ca(2+) (with EGTA present to chelate trace amounts). However, the current generated in the presence of extracellular Ca(2+) is considerably larger. 4. The pharmacological profiles for the activation of the non-selective cation current and the K(+) current are similar, suggesting that the same P2Z-like receptor could be mediating both responses. This type of plasma membrane receptor/channel-channel coupling by a process that does not appear to involve Ca(2+) flow through the receptor/channel or a subsequent membrane potential change may be representative of a new class of signalling mechanisms. PMID- 11432993 TI - Effects of membrane cholesterol manipulation on excitation-contraction coupling in skeletal muscle of the toad. AB - 1. Single mechanically skinned fibres and intact bundles of fibres from the twitch region of the iliofibularis muscle of cane toads were used to investigate the effects of membrane cholesterol manipulation on excitation-contraction (E-C) coupling. The cholesterol content of membranes was manipulated with methyl-beta cyclodextrin (MbetaCD). 2. In mechanically skinned fibres, depletion of membrane cholesterol with MbetaCD caused a dose- and time-dependent decrease in transverse tubular (t)-system depolarization-induced force responses (TSDIFRs). TSDIFRs were completely abolished within 2 min in the presence of 10 mM MbetaCD but were not affected after 2 min in the presence of a 10 mM MbetaCD-1 mM cholesterol complex. There was a very steep dependence between the change in TSDIFRs and the MbetaCD : cholesterol ratio at 10 mM MbetaCD, indicating that the inhibitory effect of MbetaCD was due to membrane cholesterol depletion and not to a pharmacological effect of the agent. Tetanic responses in bundles of intact fibres were abolished after 3-4 h in the presence of 10 mM MbetaCD. 3. The duration of TSDIFRs increased markedly soon (< 2 min) after application of 10 mM MbetaCD and 10 mM MbetaCD-cholesterol complexes, but the Ca(2+) activation properties of the contractile apparatus were minimally affected by 10 mM MbetaCD. The Ca(2+) handling abilities of the sarcoplasmic reticulum appeared to be modified after 10 min exposure to 10 mM MbetaCD. 4. Confocal laser scanning microscopy revealed that the integrity of the t-system was not compromised by either intra- or extracellular application of 10 mM MbetaCD and that a large [Ca(2+)] gradient was maintained across the t-system. 5. Membrane cholesterol depletion caused rapid depolarization of the polarized t-system as shown independently by spontaneous TSDIFRs induced by MbetaCD and by changes in the fluorescence intensity of an anionic potentiometric dye (DiBAC(4)(3)) in the presence of MbetaCD. This rapid depolarization of the t-system by cholesterol depletion was not prevented by blocking the Na(+) channels with TTX (10 microM) or the L-type Ca(2+) channels with Co(2+) (5 mM). 6. The results demonstrate that cholesterol is important for maintaining the functional integrity of the t-system and sarcoplasmic reticulum, probably by having specific effects on different membrane proteins that may be directly or indirectly involved in E-C coupling. PMID- 11432994 TI - The use of the indicator fluo-5N to measure sarcoplasmic reticulum calcium in single muscle fibres of the cane toad. AB - 1. Single fibres from the lumbrical muscles of the cane toad (Bufo marinus) were incubated in fluo-5N AM for 2 h at 35 degrees C in order to load the indicator into the sarcoplasmic reticulum. Fluo-5N is a low-affinity calcium indicator (K(Ca) 90 microM). Successful sarcoplasmic reticulum (SR) loading was indicated by a fluorescence signal that declined during contraction. 2. Confocal microscopy showed that the dye loaded principally in lines perpendicular to the long axis of the fibre that repeated each sarcomere. This is consistent with much of the dye residing in the SR. 3. To establish the site of loading, fibres were exposed to 30 mM caffeine in the presence of 20 microM 2,5-di(tert-butyl)1,4-hydroquinone (TBQ, an SR pump inhibitor) which should release most Ca(2+) from the SR; this procedure reduced the fluorescence to 46 +/- 4 % of the control value. To determine how much indicator was in the myoplasm, fibres were exposed to 100 microg ml(-1) saponin which permeabilizes the surface membrane; saponin treatment reduced the fluorescence to 51 +/- 2 % of the control value. 4. During maximally activated tetani (100 Hz stimulation rate, 22 degrees C) the component of signal from the SR declined by 33 +/- 4 %. During relaxation the SR signal recovered in two phases with time constants of 0.38 +/- 0.14 s and 10.1 +/- 1.7 s. Partially activated tetani (30 Hz stimulation rate) showed a smaller SR signal. Application of the SR Ca(2+) pump inhibitor TBQ slowed the rate of recovery of the SR signal. 5. Muscle fatigue was produced by repeated short tetani until tension was reduced to 50 %. The SR signal during the periods between tetani declined steadily and the SR Ca(2+) signal was eventually reduced to 71 +/- 8 % of the control signal. This signal recovered in two phases when the muscle was rested. An initial phase had a time constant of 1.7 +/- 0.2 s so that by 20 s of recovery the SR Ca(2+) signal was 86 +/- 7 % of control; the second phase was slower and by 5 min the SR Ca(2+) signal was back to control values (98 +/- 5 % control). In addition the magnitude of the SR signal decline associated with each tetanus (Delta[Ca(2+)](SR)) declined monotonically throughout fatigue and returned to control after 5 min recovery. 6. This approach can monitor the SR Ca(2+) concentration in normally functioning muscle fibres with good time resolution. The method confirms other approaches that show that the free Ca(2+) available for release in the SR declines during fatigue. This reduction in [Ca(2+)](SR) will contribute to the failure of Ca(2+) delivery to the myofilaments which is an important cause of muscle fatigue. PMID- 11432995 TI - Spontaneous miniature outward currents in mechanically dissociated rat Meynert neurons. AB - 1. Spontaneous miniature outward currents (SMOCs) were observed in mechanically dissociated rat Meynert neurons using nystatin perforated patch recordings under voltage-clamp conditions. 2. SMOCs were blocked by apamin, a selective blocker of small conductance Ca(2+)-activated K(+) (SK) channels, but not by blockers for other types of Ca(2+)-activated K(+) channel. 3. Ryanodine (10-100 microM) reduced both the amplitude and frequency of SMOCs. Caffeine (1 mM) increased the SMOC frequency. Blockers of the sarco/endoplasmic reticulum Ca(2+)-ATPase completely abolished SMOCs, indicating a requirement for functioning sarco/endoplasmic reticulum (SR/ER) Ca(2+) stores. 4. Both Cd(2+)-containing and Ca(2+)-free solutions partially inhibited SMOC frequency, a result which suggests that Ca(2+) influx contributes to, but is not essential for, SMOC generation. 5. Thus, SMOCs are SK currents linked to ryanodine- and caffeine-sensitive SR/ER Ca(2+) stores, and are only indirectly influenced by extracellular Ca(2+) influx. The development of this new, minimally invasive mechanical dissociation method has revealed that SMOCs are common in native CNS neurons. PMID- 11432996 TI - Local response of L-type Ca(2+) current to nitric oxide in frog ventricular myocytes. AB - 1. The regulation of L-type Ca(2+) current (I(Ca)) by the two nitric oxide (NO) donors sodium nitroprusside (SNP, 1 microM to 1 mM) and (+/-)-S-nitroso-N acetylpenicillamine (SNAP, 3 or 10 microM) was investigated in frog ventricular myocytes using double voltage clamp and double-barrelled microperfusion techniques. 2. SNP and SNAP depressed the isoprenaline (ISO, 10-100 nM)- or forskolin (FSK, 1 microM)-mediated stimulation of I(Ca) via cGMP activation of the cGMP-stimulated phosphodiesterase (PDE2). Complete inhibition of the ISO (100 nM) response was observed at 1 mM SNP. 3. When SNP was applied locally, i.e. to one-half of the cell, and ISO to the whole cell, the response of I(Ca) to ISO was strongly antagonized in the cell half exposed to SNP (up to 100 % inhibition at 1 mM SNP) but a relatively small depression was observed in the other half of the cell (only 20 % inhibition at 1 mM SNP). 4. The NO scavenger 2-(4-carboxyphenyl) 4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (carboxy-PTIO, 1 mM) reversed the local effect of SNAP (3 microM) on FSK-stimulated I(Ca) when applied to the same side as the NO donor, but had no effect when applied to the other side of the cell. 5. A local application of erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA, 30 microM), a selective inhibitor of PDE2, fully reversed the local effect of SNP (100 microM) or SNAP (10 microM) on I(Ca) but had no effect on the distant response. 6. When EHNA was applied on the distant side, with SNP (1 mM) and ISO (100 nM) applied locally, the distant effect of SNP was fully reversed. 7. Our results demonstrate that in frog ventricular myocytes stimulation of guanylyl cyclase by NO leads to a strong local depletion of cAMP near the L-type Ca(2+) channels due to activation of PDE2, but only to a modest reduction of cAMP in the rest of the cell. This may be explained by the existence of a tight microdomain between L-type Ca(2+) channels and PDE2. PMID- 11432997 TI - Two distinct oscillatory states determined by the NMDA receptor in rat inferior olive. AB - 1. The effects of N-methyl-D-aspartate (NMDA) receptor activation and blockade on subthreshold membrane potential oscillations of inferior olivary neurones were studied in brainstem slices from 12- to 21-day-old rats. 2. Dizocilpine (MK-801), a non-competitive NMDA antagonist, at 1-45 microM abolished spontaneous subthreshold oscillations, without affecting membrane potential, input resistance, or the low-threshold calcium current, I(T). Ketamine (100 microM), a non-competitive NMDA antagonist, and L-689,560 (20 microM), an antagonist at the glycine site of the NMDA receptor, also abolished the oscillations, while the competitive non-NMDA antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 20-50 microM) had no effect. 3. NMDA (100 microM) induced 4.1 Hz subthreshold oscillations and reversibly depolarized olivary neurones by 13.7 mV. In contrast, 10 microM alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and 20 microM kainic acid depolarized the membrane equivalently but did not induce oscillations. 4. Both NMDA-induced and spontaneous subthreshold oscillations were unaffected by 1 microM tetrodotoxin and were prevented by substituting extracellular calcium with cobalt. 5. Removing magnesium from the perfusate did not affect spontaneous subthreshold oscillations but did prevent NMDA-induced oscillations. 6. NMDA-induced oscillations were resistant to 50 microM mibefradil, an I(T) blocker, in contrast to spontaneous oscillations. Both oscillations were inhibited by 20 microM nifedipine, an L-type calcium channel antagonist, and 200 nM omega-agatoxin IVA, a P-type calcium channel blocker. Bay K 8644 (10 microM), an L-type Ca(2+) agonist, significantly enhanced the amplitude of both spontaneous and NMDA-induced oscillations. 7. The data indicate that NMDA receptor activation induces olivary neurones to manifest high amplitude membrane potential oscillations in part mediated by L- and P- but not T-type calcium currents. Moreover, the data demonstrate that NMDA receptor currents are necessary for generation of spontaneous subthreshold oscillations in the inferior olive. PMID- 11432998 TI - Selective inhibition of local excitatory synaptic transmission by serotonin through an unconventional receptor in the CA1 region of rat hippocampus. AB - 1. The modulation of synaptic transmission by serotonin (5-HT) was studied using whole-cell voltage-clamp and sharp-electrode current-clamp recordings from CA1 pyramidal neurones in transverse rat hippocampal slices in vitro. 2. With GABA(A) receptors blocked, polysynaptic transmission evoked by stratum radiatum stimulation was inhibited by submicromolar concentrations of 5-HT, while monosynaptic excitatory transmission and CA1 pyramidal neurone excitability were unaffected. The effect persisted following pharmacological blockade of 5-HT(1A) and 5-HT(4) receptors, which directly affect CA1 pyramidal neurone excitability. 3. Concentration-response relationships for 5-HT were determined in individual neurones; the EC(50) values for block of polysynaptic excitation and inhibition by 5-HT were approximately 230 and approximately 160 nM, respectively. The 5-HT receptor type responsible for the observed effect does not fall easily into the present classification of 5-HT receptors. 4. 5-HT inhibition of polysynaptic EPSCs persisted following complete block of GABAergic transmission and in CA1 minislices, ruling out indirect effects through interneurones and non-CA1 pyramidal neurones, respectively. 5. Monosynaptic EPSCs evoked by stimulation of CA1 afferent pathways appeared to be unaffected by 5-HT. Monosynaptic EPSCs evoked by stimulation of the alveus, which contains CA1 pyramidal neurone axons, were partially inhibited by 5-HT. 6. We conclude that 5-HT inhibited synaptic transmission by acting at local recurrent collaterals of CA1 pyramidal neurones. This may represent an important physiological action of 5-HT in the hippocampus, since it occurs over a lower concentration range than the 5-HT effects reported so far. PMID- 11432999 TI - Niflumic acid modulates uncoupled substrate-gated conductances in the human glutamate transporter EAAT4. AB - 1. The effects of niflumic acid on the substrate-gated currents mediated by the glutamate transporter EAAT4 expressed in Xenopus laevis oocytes were examined using radiolabelled substrate flux measurements and two-electrode voltage clamp techniques. 2. Niflumic acid significantly enhanced the substrate-gated currents in EAAT4, without affecting the affinity of the substrates towards EAAT4. At a concentration of 300 microM, niflumic acid caused a 19 +/- 5 % reduction in L [(3)H]glutamate uptake and no significant effect on the uptake of DL [(3)H]aspartate. Thus, enhancement of the substrate-gated currents in EAAT4 does not correlate with the rate of substrate transport and suggests that the niflumic acid-induced currents are not thermodynamically coupled to the transport of substrate. 3. Niflumic acid and arachidonic acid co-applied with substrate to EAAT4-expressing oocytes had similar functional consequences. However, niflumic acid still enhanced the L-glutamate-gated current to the same extent in the presence and absence of a saturating dose of arachidonic acid, which suggests that the sites of action of the two compounds are distinct. 4. The EAAT4-mediated currents for the two substrates, L-glutamate and L-aspartate, were not enhanced equally by addition of the same dose of niflumic acid and the ionic composition of the niflumic acid-induced currents was not the same for the two substrates. Protons carry the L-glutamate-gated niflumic acid-induced current and both protons and chloride ions carry the L-aspartate-gated niflumic acid-induced current. 5. These results show that niflumic acid can be used to probe the functional aspects of EAAT4 and that niflumic acid and other non-steroid anti inflammatory drugs could be used as the basis for the development of novel modulators of glutamate transporters. PMID- 11433000 TI - Group II and III metabotropic glutamate receptors contribute to different aspects of visual response processing in the rat superior colliculus. AB - 1. Neurones in the superior colliculus (SC) respond to novel sensory stimuli and response habituation is a key feature of this. It is known that both ionotropic and metabotropic glutamate (mGlu) receptors participate in visual responses of superficial SC neurones. A feature of Group II and Group III mGlu receptors is that they may modulate specific neural pathways, possibly via presynaptic mechanisms. However, less is known about how this may relate to functions of systems in whole animals. We have therefore investigated whether these receptors affect specific attributes of visual responses in the superficial SC. 2. Recordings were made from visually responsive neurones in anaesthetised rats, and agonists and antagonists of Group II and III mGlu receptors were applied iontophoretically at the recording site. 3. We found that application of the Group III metabotropic glutamate receptor agonist L-2-amino-4-phosphonobutyric acid (L-AP4) produced an increase in visual response habituation, whilst Group III antagonists decreased habituation. These effects were independent of the response habituation mediated via GABA(B) receptors. In contrast, modulation of Group II mGlu receptors with the specific agonist LY354740 or the antagonist LY341495 did not affect response habituation, although these compounds did modulate visual responses. This suggests a specific role for Group III mGlu receptors in visual response habituation. 4. The magnitude of Group II effects was smaller during presentation of low contrast stimuli compared with high contrast stimuli. This suggests that activation of Group II receptors may be activity dependent and that these receptors can translate this into a functional effect in adapting to high contrast stimuli. PMID- 11433001 TI - Responses to prolonged odour stimulation in frog olfactory receptor cells. AB - 1. The suction pipette technique was used to record receptor current and spiking responses from isolated frog olfactory receptor cells during prolonged odour stimuli. 2. The majority (70 %) of cells displayed 'oscillatory' responses, consisting of repeated bursts of spikes accompanied by regular increases in receptor current. The period of this oscillation varied from 3.5 to 12 s in different cells. The remaining cells responded either with a 'transient' burst of spikes at the onset of stimulation (10 %), or by 'sustained' firing throughout the odour stimulus (20 %). 3. In cells with oscillatory responses, the Ca(2+) activated Cl(-) channel blocker niflumic acid prolonged the period of oscillation only slightly, despite a 3.8-fold decrease in the receptor current. A 3-fold reduction in the external Cl(-) concentration nearly doubled the receptor current, but had little effect on the oscillation period. These results imply that the majority of the receptor current underlying these oscillatory responses is carried by the Ca(2+)-activated Cl(-) conductance, suggesting that the intracellular Ca(2+) concentration oscillates also. 4. In cells with oscillatory responses, the period of oscillation was prolonged 1.5-fold when stimulated in a low-Na(+) solution designed to incapacitate Na(+)-Ca(2+) exchange, irrespective of whether Na(+) was replaced by permeant Li(+) or impermeant choline. The dependence of the oscillation period upon external Na(+) suggests that it may be governed by the dynamics of Ca(2+) extrusion via Na(+)-Ca(2+) exchange. 5. Exposure to the membrane-permeable cyclic nucleotide analogue CPT-cAMP evoked a sustained rather than an oscillatory response even in cells with oscillatory responses to odour. The inability of CPT-cAMP to evoke an oscillatory response suggests that the cAMP concentration is likely to oscillate also. 6. Perforated patch recordings revealed that oscillatory responses could only be evoked when the membrane potential was free to change, but not when it was clamped near the resting potential. Since substantial changes in Ca(2+)-activated Cl(-) current, and hence odour-induced depolarisation, had little effect upon the period of oscillation, changes in membrane potential are suggested to play only a permissive role in these oscillatory responses. 7. These results are interpreted in terms of the coupled oscillation of Ca(2+) and cyclic nucleotide concentrations within the olfactory cilia during prolonged odour stimulation. PMID- 11433002 TI - Release of ATP by a human retinal pigment epithelial cell line: potential for autocrine stimulation through subretinal space. AB - 1. Stimulation of purinergic receptors on retinal pigment epithelial (RPE) cells can increase the rate of fluid transport or decrease phagocytosis. This study aims to: determine whether the purine ATP can be released from RPE cells, begin probing the mechanism of any release and test whether cells degrade ATP extracellularly. 2. ATP release was monitored from cultured human ARPE-19 cells with the luciferin-luciferase assay. Biphasic release of ATP was triggered by basic fibroblast growth factor (bFGF), by the pyrimidine uridine triphosphate (UTP) and by hypotonicity. 3. The Cl(-) channel blocker 5-nitro-2-(3 phenylpropylamino)-benzoate (NPPB) inhibited release of ATP, suggesting that release was associated with Cl(-) channels. 4. Elevating intracellular Ca(2+) directly with ionomycin was insufficient to trigger ATP release. 5. UTP induced a biphasic elevation in intracellular Ca(2+). NPPB inhibited the second phase, suggesting autostimulation by released ATP. 6. Cells grown on permeable supports showed apical release of ATP, analogous to release into subretinal space in vivo. 7. The presence of ecto-ATPases on ARPE-19 cell membranes was suggested by the degradation of ATP added to intact cells. 8. Phagocytosis of fluorescent beads was inhibited by ATP, but the ecto-5'-nucleotidase inhibitor alpha, beta methylene ADP prevented this, suggesting that inhibition was mediated by extracellular conversion of ATP to adenosine. 9. These results suggest that growth factors, pyrimidines and changes in tonicity could trigger ATP release into subretinal space. The levels of ATP released may be capable of autocrine stimulation of ATP receptors, while conversion to adenosine by ecto-enzymes could alter phagocytosis. PMID- 11433003 TI - Dynamic and steady-state light adaptation of mouse rod photoreceptors in vivo. AB - 1. Electroretinographic (ERG) methods were used to investigate the effects of background illumination on the responses of mouse rod photoreceptors in vivo. A paired-flash procedure, involving the recording and analysis of the ERG a-wave response to a bright probe flash presented after a brief test flash, was used to derive the rod response to the test flash in steady background light. A related, step-plus-probe procedure was used to derive the step response of the rods to backgrounds of defined strength. 2. Steady background light produced a maintained derived response that was graded with background strength. Determinations of the full time course of the derived weak-flash response in steady background light, and of the effect of background strength on the flash response at fixed post-test flash times, showed that moderate backgrounds reduce the peak amplitude and duration of the flash response. 3. The response to stepped onset of an approximately half-saturating background (1.2 sc cd m(-2)) exhibited a gradual rise over the first 200-300 ms, and an apparent subsequent relaxation to plateau amplitude within 1 s after background onset. Determinations of normalized amplitudes of the derived response to a test flash presented at 50 or 700 ms after background onset indicated substantial development of background-induced shortening of the test flash response within this 1 s period. These findings indicate a time scale of approximately 1 s or less for the near-completion of light adaptation at this background strength. 4. Properties of the derived response to a stepped background and to test flashes presented in steady background light are in general agreement with photocurrent data obtained from mammalian rods in vitro and suggest that the present results describe, to good approximation, the in vivo desensitization of mouse rods by background light. PMID- 11433004 TI - Time course of the flash response of dark- and light-adapted human rod photoreceptors derived from the electroretinogram. AB - 1. The a-wave of the electroretinogram was recorded from human subjects with normal vision, using a corneal electrode and ganzfeld stimulation. We applied the paired-flash technique, in which an intense 'probe' flash was delivered at different times after a 'test' flash. The amplitude of the probe-flash response provided a measure of the circulating current remaining at the appropriate time after the test flash. 2. We extended previous methods by measuring not at a fixed time, but at a range of times after the probe flash, and then calculating the ratio of the 'test-plus-probe' response to the 'probe-alone' response, as a function of time. 3. Under dark-adapted conditions the rod response derived by the paired-flash technique (in response to a relatively dim test flash) peaked at ca 120 ms, with a fractional sensitivity at the peak of ca 0.1 Td(-1) s(-1). 4. As reported previously, background illumination reduced the maximal response, reflecting a reduction in rod circulating current. In addition, it shortened the time to peak (to ca 70 ms at an intensity of 170 Td), and reduced the flash sensitivity measured at the peak. The flash sensitivity declined approximately according to Weber's Law, with a 10-fold reduction occurring at an intensity of 100-200 Td. We could not reliably measure responses at significantly higher background intensities because the circulating current became so small. 5. In order to investigate the phototransduction process after correction for response compression, we expressed the derived response as a fraction of the maximal response that could be elicited in the presence of the background. The earliest rising phase of this 'fractional response per unit intensity' was little affected by background illumination, suggesting that the amplification constant of transduction was unaltered by light adaptation. PMID- 11433005 TI - Modulation of adenosine transport by insulin in human umbilical artery smooth muscle cells from normal or gestational diabetic pregnancies. AB - 1. Adenosine transport was measured in human cultured umbilical artery smooth muscle cells, isolated from non-diabetic or gestational diabetic pregnancies, under basal conditions and after pretreatment in vitro with insulin. 2. Adenosine transport in non-diabetic smooth muscle cells was significantly increased by insulin (half-maximal stimulation at 0.33 +/- 0.02 nM, 8 h) and characterized by a higher maximal rate (V(max)) for nitrobenzylthioinosine (NBMPR)-sensitive (es) saturable nucleoside transport (17 +/- 5 vs. 52 +/- 12 pmol (microg protein)(-1) min(-1), control vs. insulin, respectively) and maximal binding sites (B(max)) for [(3)H]NBMPR (0.66 +/- 0.07 vs. 1.1 +/- 0.1 fmol (microg protein)(-1), control vs. insulin, respectively), with no significant changes in Michaelis-Menten (K(m)) and dissociation (K(d)) constants. 3. In contrast, in smooth muscle cells from diabetic pregnancies, where the values of V(max) for adenosine transport (59 +/- 4 pmol (microg protein)(-1) min(-1)) and B(max) for [(3)H]NBMPR binding (1.62 +/- 0.16 fmol (microg protein)(-1)) were significantly elevated by comparison with non-diabetic cells, insulin treatment (1 nM, 8 h) reduced the V(max) for adenosine transport and B(max) for [(3)H]NBMPR binding to levels detected in non diabetic cells. 4. In non-diabetic cells, the stimulatory effect of insulin on adenosine transport was mimicked by dibutyryl cGMP (100 nM) and reduced by inhibitors of phosphatidylinositol 3-kinase (10 nM wortmannin), nitric oxide synthase (100 microM N (G)-nitro-L-arginine methyl ester, L-NAME) or protein synthesis (1 microM cycloheximide), whereas inhibition of adenylyl cyclase (100 microM SQ-22536) had no effect. 5. Wortmannin or SQ-22536, but not L-NAME or cycloheximide, attenuated the inhibitory action of insulin on the diabetes induced stimulation of adenosine transport. 6. Protein levels of inducible NO synthase (iNOS) were similar in non-diabetic and diabetic cells, but were increased by insulin (1 nM, 8 h) only in non-diabetic smooth muscle cells. 7. Our results suggest that adenosine transport via the es nucleoside transporter is modulated differentially by insulin in either cell type. Insulin increased adenosine transport in non-diabetic cells via NO and cGMP, but inhibited the diabetes-elevated adenosine transport via activation of adenylyl cyclase, suggesting that the biological actions of adenosine may be altered under conditions of sustained hyperglycaemia in uncontrolled diabetes. PMID- 11433006 TI - Intraganglionic laminar endings are mechano-transduction sites of vagal tension receptors in the guinea-pig stomach. AB - 1. Distension-sensitive vagal afferent fibres from the cardiac region of the guinea-pig stomach were recorded extracellularly, then filled with biotinamide, using an anterograde tracing technique. 2. Most of the stretch-sensitive units of the guinea-pig stomach (41 out of 47; number of animals N = 26) had low thresholds (less than 1 mm) to circumferential stretch and showed slow adaptation. Twenty of these units fired spontaneously under resting conditions (mean: 1.9 +/- 0.3 Hz, n = 20, N = 14). 3. Adaptation of firing during slow or maintained stretch correlated closely with accommodation of intramural tension, but tension-independent adaptation was also present. 4. Nicardipine (3 microM) with hyoscine (3 microM) reduced stretch-evoked firing of gastric vagal afferents, by inhibiting smooth muscle contraction. Gadolinium (1 mM) blocked distension-evoked firing. 5. Focal stimulation of the stomach muscle wall with a von Frey hair (0.4 mN) identified one to six punctate receptive fields in each low threshold vagal distension-sensitive afferent. These were marked on the serosal surface of the stomach wall. 6. Anterograde filling of recorded nerve trunks revealed intraganglionic laminar endings (IGLEs) within 142 +/- 34 microm (n = 38; N = 10) of marked receptive fields. The mean distance from randomly generated sites to the nearest IGLE was significantly greater (1500 +/- 48 microm, n = 380, N = 10, P < 0.0001). Viscerofugal nerve cell bodies, intramuscular arrays and varicose axons were not associated with receptive fields. The results indicate that IGLEs are the mechanotransduction sites of low threshold, slowly adapting vagal tension receptors in the guinea-pig upper stomach. PMID- 11433007 TI - Adrenaline increases skeletal muscle glycogenolysis, pyruvate dehydrogenase activation and carbohydrate oxidation during moderate exercise in humans. AB - 1. To evaluate the role of adrenaline in regulating carbohydrate metabolism during moderate exercise, 10 moderately trained men completed two 20 min exercise bouts at 58 +/- 2 % peak pulmonary oxygen uptake (V(O2,peak)). On one occasion saline was infused (CON), and on the other adrenaline was infused intravenously for 5 min prior to and throughout exercise (ADR). Glucose kinetics were measured by a primed, continuous infusion of 6,6-[(2)H]glucose and muscle samples were obtained prior to and at 1 and 20 min of exercise. 2. The infusion of adrenaline elevated (P < 0.01) plasma adrenaline concentrations at rest (pre-infusion, 0.28 +/- 0.09; post-infusion, 1.70 +/- 0.45 nmol l(-1); means +/- S.E.M.) and this effect was maintained throughout exercise. Total carbohydrate oxidation increased by 18 % and this effect was due to greater skeletal muscle glycogenolysis (P < 0.05) and pyruvate dehydrogenase (PDH) activation (P < 0.05, treatment effect). Glucose rate of appearance was not different between trials, but the infusion of adrenaline decreased (P < 0.05, treatment effect) skeletal muscle glucose uptake in ADR. 3. During exercise muscle glucose 6-phosphate (G-6-P) (P = 0.055, treatment effect) and lactate (P < 0.05) were elevated in ADR compared with CON and no changes were observed for pyruvate, creatine, phosphocreatine, ATP and the calculated free concentrations of ADP and AMP. 4. The data demonstrate that elevated plasma adrenaline levels during moderate exercise in untrained men increase skeletal muscle glycogen breakdown and PDH activation, which results in greater carbohydrate oxidation. The greater muscle glycogenolysis appears to be due to increased glycogen phosphorylase transformation whilst the increased PDH activity cannot be readily explained. Finally, the decreased glucose uptake observed during exercise in ADR is likely to be due to the increased intracellular G-6-P and a subsequent decrease in glucose phosphorylation. PMID- 11433008 TI - Middle cerebral artery blood velocity is reduced with hyperthermia during prolonged exercise in humans. AB - 1. In the present study we examined the effect of hyperthermia on the middle cerebral artery mean blood velocity (MCA V(mean)) during prolonged exercise. We predicted that the cerebral circulation would be impaired when hyperthermia is present during exercise and assumed that this could be observed as a reduced MCA V(mean). 2. Eight endurance trained men (maximum oxygen uptake (V(O2,max)) 70 +/- 1 ml min(-1) kg(-1) (mean +/- S.E.M.)) performed two exercise trials at 57 % of V(O2,max) on a cycle ergometer in a hot (40 degrees C; hyperthermic trial) and in a thermoneutral environment (18 degrees C; control trial). In the hyperthermic trial, the oesophageal temperature increased throughout the exercise period reaching a peak value of 40.0 +/- 0.1 degrees C at exhaustion after 53 +/- 4 min of exercise. In the control trial, exercise was maintained for 1 h without any signs of fatigue and with core temperature stabilised at 37.8 +/- 0.1 degrees C after approximately 15 min of exercise. 3. Concomitant with the development of hyperthermia, MCA V(mean) declined by 26 +/- 3 % from 73 +/- 4 cm s(-1) at the beginning of exercise to 54 +/- 4 cm s(-1) at exhaustion (P < 0.001). In contrast, MCA V(mean) remained unchanged at 70-72 cm s(-1) throughout the 1 h control trial. 4. When individually determined regression lines for MCA V(mean) and arterial carbon dioxide pressure (P(a,CO2)) obtained during preliminary exercise tests were used to account for the differences in P(a,CO2) between the hyperthermic and control trial, it appeared that more than half of the reduction in MCA V(mean) (56 +/- 8 %) was related to a hyperventilation-induced drop in P(a,CO2). Declining cardiac output and arterial blood pressure accounted for the remaining part of the hyperthermia-induced reduction in MCA V(mean). 5. The present results demonstrate that the development of hyperthermia during prolonged exercise is associated with a marked reduction in MCA V(mean). PMID- 11433009 TI - Regular endurance exercise induces expansive arterial remodelling in the trained limbs of healthy men. AB - 1. In experimental animals chronic elevations in arterial blood flow increase the lumen diameter and reduce the intima-media thickness (IMT) of the arterial segment involved. We determined whether intermittent elevations in active muscle blood flow associated with regular aerobic leg exercise induced such expansive arterial remodelling in the common femoral artery of humans. 2. In the cross sectional study 53 sedentary (47 +/- 2 years) and 55 endurance exercise-trained (47 +/- 2 years) men were studied. Common femoral artery lumen diameter (B-mode ultrasound) was 7 % greater (9.62 +/- 0.12 vs. 9.03 +/- 0.13 mm), and femoral IMT (0.46 +/- 0.02 vs. 0.55 +/- 0.02 mm) and IMT/lumen ratio were 16-21 % smaller in the endurance-trained men (all P < 0.001). Basal femoral artery blood flow (duplex ultrasound) was not different, shear stress tended to be lower (P = 0.08), and mean femoral tangential wall stress was 30 % higher in the endurance trained men (P < 0.001). 3. In the intervention study 22 men (51 +/- 2 years) were studied before and after 3 months of regular aerobic leg exercise (primarily walking). After training, the femoral diameter increased by 9 % (8.82 +/- 0.18 vs. 9.60 +/- 0.20 mm), and IMT (0.65 +/- 0.05 vs. 0.56 +/- 0.05 mm) and the IMT/lumen ratio were approximately 15-20 % smaller (all P < 0.001). Basal femoral blood flow and shear stress were not different after training, whereas the mean femoral tangential wall stress increased by 31 %. The changes in arterial structure were not related to changes in risk factors for atherosclerosis. 4. Our results are consistent with the concept that regular aerobic leg exercise induces expansive arterial remodelling in the femoral artery of healthy men. This adaptive process is produced by even a moderate training stimulus, is not obviously dependent on corresponding improvements in risk factors for atherosclerosis, and is robust, occurring in healthy men of different ages. PMID- 11433010 TI - Training-induced changes in peritendinous type I collagen turnover determined by microdialysis in humans. AB - 1. Acute exercise is found to increase collagen type I formation locally in peritendinous connective tissue of the Achilles' tendon in humans, as determined from changes in interstitial concentrations of collagen propeptide (PICP) and a collagen degradation product (ICTP) by the use of microdialysis catheters. However, the local collagen type I turnover response to training is unknown. 2. Nineteen young males were studied before and after 4 and 11 weeks of physical training. Microdialysis catheters with a high molecular mass cut-off value (3000 kDa), allowing the determination of PICP and ICTP, were placed in the peritendinous space ventral to the Achilles' tendon, under ultrasound guidance, in both legs. The catheters were perfused with a Ringer-acetate solution containing (3)H-labelled human type IV collagen for in vivo recovery determination (relative recovery: 79 +/- 2 %, mean +/- S.E.M.). 3. The PICP concentration in the peritendinous tissue increased in response to training (from 5 +/- 1 to 35 +/- 5 microg l(-1) (4 weeks), P < 0.05) and remained elevated throughout the training period (28 +/- 6 microg l(-1), 11 weeks). Tissue ICTP only rose transiently with training (from 2.2 +/- 0.1 to 2.8 +/- 0.2 microg l(-1) (4 weeks), P < 0.05, and 2.5 +/- 0.2 microg l(-1) (11 weeks), P > 0.05 vs. basal). Plasma PICP was unchanged whereas plasma ICTP declined by 17 % in response to training. 4. The findings indicate that physical training results in an increased turnover of collagen type I in local connective tissue of the peritendinous Achilles' region. Early in the process both synthesis and degradation are elevated, whereas later, the anabolic processes are dominating causing a net synthesis of type I collagen in tendon-related tissue in humans. PMID- 11433011 TI - Obstacle avoidance during human walking: learning rate and cross-modal transfer. AB - 1. The aim of this study was to investigate the significance of specific afferent information during motor learning. Blindfolded subjects stepped over an obstacle on a treadmill while different stimuli (acoustic (ACU), somatosensory (SOM) and light flash (LED)) signalled the approaching obstacle. The effect of the above stimuli was then evaluated and compared to full vision (VIS) locomotion. In the non-visual conditions feedback information about the performance was provided by an acoustic signal. 2. Using each of the different stimuli for information the level of subject performance was assessed by noting foot clearance and analysing both leg muscle electromyographic activity and movement trajectories during three successive runs. Each of these runs consisted of 100 steps over the obstacle. 3. The best performance at the onset of the first run was achieved during the VIS condition. When the VIS condition (run 1 + 2) was followed by ACU or SOM information or when the ACU condition (run 1 + 2) was followed by LED, little cross-modal transfer (CMT) occurred, i.e. adaptation in run 3 started again at a low level of performance. In contrast, if adaptation started with ACU stimuli followed by SOM stimuli, almost full CMT occurred. The absolute level of performance achieved after the second or third runs was similar in the VIS and non-VIS conditions. 4. In conclusion, the course of motor learning depends on specific afferent information, and feedforward control has a special influence on the performance only at the onset of the experiment but not on the rate of learning. The fact that little CMT occurs from visual to non-visual stimuli and from ACU to LED suggests that visual afferent input is processed in a different way to non-visual stimuli. PMID- 11433013 TI - A sensitive, single-tube assay to measure the enzymatic activities of influenza RNA polymerase and other poly(A) polymerases: application to kinetic and inhibitor analysis. AB - We describe a fast and robust new assay format to measure poly(A) polymerase (PAP) activity in a microtiter plate format. The new assay principle uses only natural nucleotide triphosphates and avoids a labour-intensive filtration step. A coupled enzymatic system combining PAP and reverse transcriptase forms the basis of the assay. The PAP generates a poly(A) tail on a RNA substrate and the reverse transcriptase is used to quantify the polyadenylated RNA by extension of a biotinylated oligo-dT primer. We demonstrate the principle of the assay using influenza virus RNA polymerase and yeast PAP as examples. A specific increase in the K(m) value for ATP and the observation of burst kinetics in the polyadenylation dependent, but not in the polyadenylation independent, assay suggest that a rate limiting step of influenza polymerase activity occurs after transcription elongation. Yeast PAP was used to validate the assay as an example of a template independent PAP. The new yeast PAP assay was approximately 100-fold more sensitive than the conventional TCA precipitation assay for yeast PAP, but the kinetic analysis of the PAP reaction gave similar results in both assays. The two enzymes show important differences with respect to inhibition by 3'-deoxy ATP. Whereas the K(i) value for 3'-deoxy-ATP (105-117 microM) is similar to the K(m) value for ATP (186 microM) in the case of influenza RNA polymerase, the K(i) value for 3'-deoxy-ATP (0.4-0.6 microM) is approximately 100-fold lower than the K(m) value for ATP (50 microM) in the case of yeast PAP. PMID- 11433012 TI - Comparison of the RNA polymerase III transcription machinery in Schizosaccharomyces pombe, Saccharomyces cerevisiae and human. AB - Multi-subunit transcription factors (TF) direct RNA polymerase (pol) III to synthesize a variety of essential small transcripts such as tRNAs, 5S rRNA and U6 snRNA. Use by pol III of both TATA-less and TATA-containing promoters, together with progress in the Saccharomyces cerevisiae and human systems towards elucidating the mechanisms of actions of the pol III TFs, provides a paradigm for eukaryotic gene transcription. Human and S.cerevisiae pol III components reveal good general agreement in the arrangement of orthologous TFs that are distributed along tRNA gene control elements, beginning upstream of the transcription initiation site and extending through the 3' terminator element, although some TF subunits have diverged beyond recognition. For this review we have surveyed the Schizosaccharomyces pombe database and identified 26 subunits of pol III and associated TFs that would appear to represent the complete core set of the pol III machinery. We also compile data that indicate in vivo expression and/or function of 18 of the fission yeast proteins. A high degree of homology occurs in pol III, TFIIIB, TFIIIA and the three initiation-related subunits of TFIIIC that are associated with the proximal promoter element, while markedly less homology is apparent in the downstream TFIIIC subunits. The idea that the divergence in downstream TFIIIC subunits is associated with differences in pol III termination related mechanisms that have been noted in the yeast and human systems but not reviewed previously is also considered. PMID- 11433014 TI - Histone variant macroH2A contains two distinct macrochromatin domains capable of directing macroH2A to the inactive X chromosome. AB - Chromatin on the inactive X chromosome (Xi) of female mammals is enriched for the histone variant macroH2A that can be detected at interphase as a distinct nuclear structure referred to as a macro chromatin body (MCB). Green fluorescent protein tagged and Myc epitope-tagged macroH2A readily form an MCB in the nuclei of transfected female, but not male, cells. Using targeted disruptions, we have identified two macrochromatin domains within macroH2A that are independently capable of MCB formation and association with the Xi. Complete removal of the non histone C-terminal tail does not reduce the efficiency of association of the variant histone domain of macroH2A with the Xi, indicating that the histone portion alone can target the Xi. The non-histone domain by itself is incapable of MCB formation. However, when directed to the nucleosome by fusion to core histone H2A or H2B, the non-histone tail forms an MCB that appears identical to that of the endogenous protein. Mutagenesis of the non-histone portion of macroH2A localized the region required for MCB formation and targeting to the Xi to an approximately 190 amino acid region. PMID- 11433015 TI - Analysis of the open region of RNA polymerase II transcription complexes in the early phase of elongation. AB - The RNA polymerase II (pol II) transcription complex undergoes a structural transition around registers 20-25, as indicated by ExoIII footprinting analyses. We have employed a highly purified system to prepare pol II complexes stalled at very precise positions during the initial stage of transcript elongation. Using potassium permanganate we analyzed the open region ('transcription bubble') of complexes stalled between registers 15 and 35. We found that from register 15 up to 25 the transcription bubble expands concomitantly with RNA synthesis. At registers 26 and 27 the bubble has a high tendency to retract at the leading edge. Addition of transcription elongation factor TFIIS re-extends the bubble to the stall site, resulting in complexes competent for transcript elongation. These findings are discussed in the light of the recently determined structures for RNA polymerases. PMID- 11433016 TI - Primer-dependent synthesis by poliovirus RNA-dependent RNA polymerase (3D(pol)). AB - Properties of poliovirus RNA-dependent RNA polymerase (3D(pol)) including optimal conditions for primer extension, processivity and the rate of dissociation from primer-template (k(off)) were examined in the presence and absence of viral protein 3AB. Primer-dependent polymerization was examined on templates of 407 or 1499 nt primed such that fully extended products would be 296 or 1388 nt, respectively. Maximal primer extension was achieved with low rNTP concentrations (50-100 microM) using pH 7 and low (<1 mM) MgCl(2) and KCl (<20 mM) concentrations. However, high activity (about half maximal) was also observed with 500 microM rNTPs providing that higher MgCl(2) levels (3-5 mM) were used. The enhancement observed with the former conditions appeared to result from a large increase in the initial level or active enzyme that associated with the primer. 3AB increased the number of extended primers at all conditions with no apparent change in processivity. The k(off) values for the polymerase bound to primer-template were 0.011 +/- 0.005 and 0.037 +/- 0.006 min(-1) (average of four or more experiments +/- SD) in the presence or absence of 3AB, respectively. The decrease in the presence of 3AB suggested an enhancement of polymerase binding or stability. However, binding was tight even without 3AB, consistent with the highly processive (at least several hundred nucleotides) nature of 3D(pol). The results support a mechanism whereby 3AB enhances the ability of 3D(pol) to form a productive complex with the primer-template. Once formed, this complex is very stable resulting in highly processive synthesis. PMID- 11433017 TI - Translesional synthesis on DNA templates containing the 2'-deoxyribonolactone lesion. AB - A site-specifically modified oligonucleotide containing a single 2' deoxyribonolactone lesion was used as a template for primer extension reactions catalyzed by M-MuLV reverse transcriptase (RT) and by the Klenow fragments of Escherichia coli DNA polymerase proficient (KF exo(+)) or deficient (KF exo(-)) in exonuclease activity. Analysis of the extension products in the presence of the four dNTPs or of a single dNTP showed that the M-MuLV RT was completely blocked and did not incorporate any dNMP opposite 2'-deoxyribonolactone. KF exo( ) preferentially incorporated nucleotides opposite the lesion following the frequency order dAMP > dGMP >> dTMP approximately dCMP and thus appeared to obey the 'A rule' for preferential incorporation as has been shown previously for the 2'-deoxyribose abasic site. In the sequence context examined, the primer extension by KF exo(-) appeared to be less efficient when dAMP was positioned opposite the lesion as compared with dTMP or dGMP. These two nucleotides promoted a more efficient polymerization accompanied by nucleotide deletion through misalignment incorporations. We therefore predict that the sequence context may strongly influence the translesional synthesis by KF exo(-) and thus the miscoding and mutational potential of the 2'-deoxyribonolactone in E.coli. PMID- 11433018 TI - Accumulation of H/ACA snoRNPs depends on the integrity of the conserved central domain of the RNA-binding protein Nhp2p. AB - Box H/ACA small nucleolar ribonucleoprotein particles (H/ACA snoRNPs) play key roles in the synthesis of eukaryotic ribosomes. How box H/ACA snoRNPs are assembled remains unknown. Here we show that yeast Nhp2p, a core component of these particles, directly binds RNA. In vitro, Nhp2p interacts with high affinity with RNAs containing irregular stem-loop structures but shows weak affinity for poly(A), poly(C) or for double-stranded RNAs. The central region of Nhp2p is believed to function as an RNA-binding domain, since it is related to motifs found in various RNA-binding proteins. Removal of two amino acids that shortens a putative beta-strand element within Nhp2p central domain impairs the ability of the protein to interact with H/ACA snoRNAs in cell extracts. In vivo, this deletion prevents cell viability and leads to a strong defect in the accumulation of H/ACA snoRNAs and Gar1p. These data suggest that proper direct binding of Nhp2p to H/ACA snoRNAs is required for the assembly of H/ACA snoRNPs and hence for the stability of some of their components. In addition, we show that converting a highly conserved glycine residue (G(59)) within Nhp2p central domain to glutamate significantly reduces cell growth at 30 and 37 degrees C. Remarkably, this modification affects the steady-state levels of H/ACA snoRNAs and the strength of Nhp2p association with these RNAs to varying degrees, depending on the nature of the H/ACA snoRNA. Finally, we show that the modified Nhp2p protein whose interaction with H/ACA snoRNAs is impaired cannot accumulate in the nucleolus, suggesting that only the assembled H/ACA snoRNP particles can be efficiently retained in the nucleolus. PMID- 11433019 TI - Ribosomal protein gene cluster analysis in eubacterium genomics: homology between Sinorhizobium meliloti strain 1021 and Bacillus subtilis. AB - The first whole genome sequence of a symbiotic soil bacterium, Sinorhizobium meliloti (formely named Rhizobium meliloti) strain 1021, is due in 2001. As an active participant in the European and North American consortium that has completed this work, our group has sequenced a region on the chromosome containing clusters rpoBC, str, S10, spc and alpha corresponding to 30 protein genes. The structural organization and function of these genes were compared with those of orthologs in another 15 complete eubacterial genomes available in databases. This study, involving the DNA and amino acid sequences as well as the organization of the whole region (gene order, cluster order, etc.), has shown that the phylogenetic tree resulting from a comparison of the amino acid sequence is rather similar to that derived from 16S rRNA sequence data. However, the tree achieved by aligning DNA sequences groups the organisms with a high GC content (>60% GC), while that based on a comparison of gene cluster orientation and organization reveals a greater level of correspondence between the alpha proteobacteria S.meliloti and the firmicute Bacillus subtilis. PMID- 11433020 TI - Initiation of HIV-2 reverse transcription: a secondary structure model of the RNA tRNA(Lys3) duplex. AB - Human immunodeficiency virus type 2 (HIV-2) reverse transcription is initiated from cellular tRNA(Lys3) partially annealed to the RNA viral genome at the primer binding site (PBS). This annealing involves interactions between two highly structured RNA molecules. In contrast to HIV-1, in which the reverse transcription initiation complex has been thoroughly studied, there is still little information regarding a possible model to describe the secondary structure of the template-primer complex in HIV-2. To determine whether HIV-2 RNA sequences flanking the PBS may specifically interact with the natural primer tRNA, we performed site-directed mutagenesis and enzymatic footprinting. An RNA fragment corresponding to the HIV-2 U5 RNA domain and tRNA(Lys3) were probed either in their free form or in the binary complex. Important reactivity changes to nucleases were obtained upon complex formation. In addition to the canonical contacts between the viral PBS and the 3' end acceptor stem of tRNA(Lys3), we identified two additional interacting domains: (i) the U-rich region of the anticodon loop with the A-rich sequence of the internal loop within the U5-prePBS region; (ii) nucleotides 48-54 from the TPsiC domain of tRNA(Lys3) and the 240 247 region of viral U5-RNA. In view of these experimental data and sequence comparison between different HIV-2 isolates, we propose a model for the secondary structure of the HIV-2 template-primer initiation complex. PMID- 11433021 TI - In vivo co-localisation of MBNL protein with DMPK expanded-repeat transcripts. AB - Myotonic dystrophy (DM1) is the most common form of adult muscular dystrophy and is inherited as an autosomal dominant trait. The genetic basis of DM1 is the expansion of a CTG repeat in the 3' untranslated region of a protein kinase gene (DMPK). The molecular mechanism by which this expanded repeat produces the pathophysiology of DM1 remains unknown. Transcripts from the expanded allele accumulate as foci in the nucleus of DM1 cells and it has been suggested that these transcript foci sequester cellular proteins that are required for normal nuclear function. We have investigated the role of three RNA-binding proteins, CUG-BP, hnRNP C and MBNL, as possible sequestered factors. Using a combination of indirect immunofluorescence to detect endogenous proteins and overexpression of proteins with green fluorescent protein (GFP) tags we have shown that CUG-BP and hnRNP C do not co-localise with expanded repeat foci in DM1 cell lines. However, GFP-tagged MBNL does itself form foci in DM1 cell lines and co-localises with the foci of expanded repeat transcripts. GFP-tagged MBNL does not appear as foci in non-DM1 cell lines. This work provides further support for the involvement of MBNL in DM1. PMID- 11433022 TI - Unusually wide co-factor tolerance in a metalloenzyme; divalent metal ions modulate endo-exonuclease activity in T5 exonuclease. AB - T5 5'-3' exonuclease is a member of a homologous group of 5' nucleases which require divalent metal co-factors. Structural and biochemical studies suggest that single-stranded DNA substrates thread through a helical arch or hole in the protein, thus bringing the phosphodiester backbone into close proximity with the active site metal co-factors. In addition to the expected use of Mg(2+), Mn(2+) and Co(2+) as co-factors, we found that divalent zinc, iron, nickel and copper ions also supported catalysis. Such a range of co-factor utilisation is unusual in a single enzyme. Some co-factors such as Mn(2+) stimulated the cleavage of double-stranded closed-circular plasmid DNA. Such endonucleolytic cleavage of circular double-stranded DNA cannot be readily explained by the threading model proposed for the cleavage of substrates with free 5'-ends as the hole observed in the crystal structure of T5 exonuclease is too small to permit the passage of double-stranded DNA. We suggest that such a substrate may gain access to the active site of the enzyme by a process which does not involve threading. PMID- 11433023 TI - Significantly higher activity of a cytoplasmic hammerhead ribozyme than a corresponding nuclear counterpart: engineered tRNAs with an extended 3' end can be exported efficiently and specifically to the cytoplasm in mammalian cells. AB - Hammerhead ribozymes were expressed under the control of similar tRNA promoters, localizing transcripts either in the cytoplasm or the nucleus. The tRNA(Val) driven ribozyme (tRNA-Rz; tRNA with extra sequences at the 3' end) that has been used in our ribozyme studies was exported efficiently into the cytoplasm and ribozyme activity was detected only in the cytoplasmic fraction. Both ends of the transported tRNA-Rz were characterized comprehensively and the results confirmed that tRNA-Rz had unprocessed 5' and 3' ends. Furthermore, it was also demonstrated that the activity of the exported ribozyme was significantly higher than that of the ribozyme which remained in the nucleus. We suggest that it is possible to engineer tRNA-Rz, which can be exported to the cytoplasm based on an understanding of secondary structures, and then tRNA-driven ribozymes may be co localized with their target mRNAs in the cytoplasm of mammalian cells. PMID- 11433024 TI - E2F-1 represses transcription of the human telomerase reverse transcriptase gene. AB - The ends of human chromosomes (telomeres) lose up to 200 bp of DNA per cell division. Chromosomal shortening ultimately leads to senescence and death in normal cells. Many human carcinoma lines are immortal in vitro, suggesting that these cells have a mechanism for maintaining the ends of their chromosomes. Telomerase is a ribonucleoprotein complex that synthesizes telomeric DNA onto chromosomes using its RNA component as template. Telomerase activity is found in most tumor cells, but is absent from normal cells. Little is known about how normal human cells repress telomerase (hTERT) gene expression. Mice carrying an E2F-1 null mutation develop a variety of malignant tumors, suggesting that this transcription factor has a tumor suppressor function. To determine mechanisms by which E2F-1 suppresses tumor formation, we examined the role of this transcription factor in regulation of the hTERT promoter in human cells. We identified two putative E2F-1-binding sites proximal to the transcriptional start site of the hTERT promoter. Mutation of these sites produced dramatic increases in promoter activity. Overexpression of E2F-1 but not a mutant E2F-1 repressed hTERT promoter activity in reporter gene assays. This repression was abolished by mutation of the E2F-1-binding sites in the hTERT promoter. Human cancer cell lines stably overexpressing E2F-1 exhibited decreased hTERT mRNA expression and telomerase activity. We conclude that E2F-1 has an atypical function as a transcriptional repressor of the hTERT gene in human cells. PMID- 11433025 TI - Hexamminecobalt(III)-induced condensation of calf thymus DNA: circular dichroism and hydration measurements. AB - The interaction of hexamminecobalt(III), Co(NH(3))(6)(3+), with 160 and 3000-8000 bp length calf thymus DNA has been investigated by circular dichroism, acoustic and densimetric techniques. The acoustic titration curves of 160 bp DNA revealed three stages of interaction: (i) Co(NH(3))(6)(3+) binding up to the molar ratio [Co(NH(3))(6)(3+)]/[P] = 0.25, prior to DNA condensation; (ii) a condensation process between [Co(NH(3))(6)(3+)]/[P] = 0.25 and 0.30; and (iii) precipitation after [Co(NH(3))(6)(3+)]/[P] = 0.3. In the case of 3000-8000 bp DNA only two processes were observed: (i) binding up to [Co(NH(3))(6)(3+)]/[P] = 0.3; and (ii) precipitation after this point. In agreement with earlier observations, long DNA aggregates without changes in its B-form circular dichroism spectrum, while short DNA demonstrates a positive B-->Psi transition after [Co(NH(3))(6)(3+)]/[P] = 0.25. From ultrasonic and densimetric measurements the effects of Co(NH(3))(6)(3+) binding on volume and compressibility have been obtained. The binding of Co(NH(3))(6)(3+) to both short and long DNA is characterized by similar changes in volume and compressibility calculated per mole Co(NH(3))(6)(3+): DeltaV = 9 cm(3) mol(-1) and Deltakappa = 33 x 10(-4) cm(3) mol(-1) bar(-1). The positive sign of the parameters indicates dehydration, i.e. water release from Co(NH(3))(6)(3+) and the atomic groups of DNA. This extent of water displacement would be consistent with the formation of two direct, hydrogen bonded contacts between the cation and the phosphates of DNA. PMID- 11433026 TI - hMYH cell cycle-dependent expression, subcellular localization and association with replication foci: evidence suggesting replication-coupled repair of adenine:8-oxoguanine mispairs. AB - The human MutY homolog, hMYH, is an adenine-specific DNA glycosylase that removes adenines or 2-hydroxyadenines mispaired with guanines or 8-oxoguanines. In order to prevent mutations, this activity must be directed to the newly synthesized strand and not the template strand during DNA synthesis. The subcellular localization and expression of hMYH has been studied in serum-stimulated, proliferating MRC5 cells. Using specific antibodies, we demonstrate that endogenous hMYH protein localized both to nuclei and mitochondria. hMYH in the nuclei is distinctly distributed and co-localized with BrdU at replication foci and with proliferating cell nuclear antigen (PCNA). The levels of hMYH in the nucleus increased 3- to 4-fold during progression of the cell cycle and reached maximum levels in S phase compared to early G(1). Similar results were obtained for PCNA, while there were no notable changes in expression of 8-oxoguanine glycosylase or the human MutT homolog, MTH1, throughout the cell cycle. The cell cycle-dependent expression and localization of hMYH at sites of DNA replication suggest a role for this glycosylase in immediate post-replication DNA base excision repair. PMID- 11433027 TI - E2F mediates induction of the Sp1-controlled promoter of the human DNA polymerase epsilon B-subunit gene POLE2. AB - The B-subunits of replicative DNA polymerases from Archaea to humans belong to the same protein family, suggesting that they share a common fundamental function. We report here the gene structure for the B-subunit of human DNA polymerase epsilon (POLE2), whose expression and transcriptional regulation is typical for replication proteins with some unique features. The 75 bp core promoter region, located within exon 1, contains an Sp1 element that is a critical determinant of promoter activity as shown by the luciferase reporter, electrophoretic mobility shift and DNase I footprinting assays. Two overlapping E2F elements adjacent to the Sp1 element are essential for full promoter activity and serum response. Binding sites for E2F1 and NF-1 reside immediately downstream from the core promoter region. Our results suggest that human POLE2 is regulated by two E2F-pocket protein complexes, one associated with Sp1 and the other with NF-1. So far, only one replicative DNA polymerase B-subunit gene promoter, POLA2 encoding the B-subunit of DNA polymerase alpha, has been characterized. Mitogenic activation of the POLE2 promoter by an E2F-mediated mechanism resembles that of POLA2, but the regulation of basal promoter activity is different between these two genes. PMID- 11433028 TI - Developmentally-regulated packaging of mitochondrial DNA by the HMG-box protein mtTFA during Xenopus oogenesis. AB - Mature Xenopus oocytes are highly enriched for mitochondria. The organelles are stored and partitioned to newly-arising cells during embryogenesis, when there is little mitochondrial DNA (mtDNA) replication or transcription. A previously described member of the high mobility group (HMG) family of proteins, mtTFA, has been suggested to play a role in control of mtDNA copy number. mtTFA serves as a mitochondrial transcription factor in humans and Xenopus and as an abundant mtDNA packaging protein in yeast, like its prokaryotic histone-like counterpart, HU protein. Northern blot analysis demonstrated that expression of the gene was regulated during Xenopus oogenesis and specifically peaked at stage II. Western and Southern blotting were used to quantify amounts of the protein and mtDNA, respectively, in each stage of oogenesis. mtTFA:mtDNA ratios were found to be relatively low in previtellogenic oocytes while the ratios increased markedly in mature oocytes. PMID- 11433029 TI - Measurement of steady-state kinetic parameters for DNA unwinding by the bacteriophage T4 Dda helicase: use of peptide nucleic acids to trap single stranded DNA products of helicase reactions. AB - Measurement of steady-state rates of unwinding of double-stranded oligonucleotides by helicases is hampered due to rapid reannealing of the single stranded DNA products. Including an oligonucleotide in the reaction mixture which can hybridize with one of the single strands can prevent reannealing. However, helicases bind to single-stranded DNA, therefore the additional oligonucleotide can sequester the enzyme, leading to slower observed rates for unwinding. To circumvent this problem, the oligonucleotide that serves as a trap was replaced with a strand of peptide nucleic acid (PNA). Fluorescence polarization was used to determine that a 15mer PNA strand does not bind to the bacteriophage T4 Dda helicase. Steady-state kinetic parameters of unwinding catalyzed by Dda were determined by using PNA as a trapping strand. The substrate consisted of a partial duplex with 15 nt of single-stranded DNA and 15 bp. In the presence of 250 nM substrate and 1 nM Dda, the rate of unwinding in the presence of the DNA trapping strand was 0.30 nM s(-1) whereas the rate was 1.34 nM s(-1) in the presence of the PNA trapping strand. PNA prevents reannealing of single-stranded DNA products, but does not sequester the helicase. This assay will prove useful in defining the complete kinetic mechanism for unwinding of oligonucleotide substrates by this helicase. PMID- 11433030 TI - 14-3-3tau associates with and activates the MEF2D transcription factor during muscle cell differentiation. AB - Myocyte enhancer binding factor 2 (MEF2) proteins belong to the MADS box family of transcription factors and four MEF2 proteins, MEF2A, MEF2B, MEF2C and MEF2D, have been found. MEF2 proteins have been shown to play critical roles in differentiation of muscles and neuronal tissues. How transactivational activity of MEF2 proteins is regulated is not fully understood. MEF2 proteins are activated by several kinases, including Erk5 and calcium/calmodulin-dependent kinase, and interact with repressors, including histone deacetylases 4 and 5 (HDAC4 and HDAC5) and Cabin1. During the effort to understand regulation of MEF2 activity, we identified 14-3-3tau as a MEF2D-interacting molecule by yeast two hybrid screening. We found that 14-3-3tau forms a complex with MEF2D in vivo and specifically enhances MEF2 transactivational activity. The results from transient transfection and co-precipitation experiments suggest that 14-3-3tau activates MEF2D by competitively inhibiting HDAC4 from binding to MEF2D and thereby affects muscle cell differentiation. PMID- 11433031 TI - The Bloom's and Werner's syndrome proteins are DNA structure-specific helicases. AB - BLM and WRN, the products of the Bloom's and Werner's syndrome genes, are members of the RecQ family of DNA helicases. Although both have been shown previously to unwind simple, partial duplex DNA substrates with 3'-->5' polarity, little is known about the structural features of DNA that determine the substrate specificities of these enzymes. We have compared the substrate specificities of the BLM and WRN proteins using a variety of partial duplex DNA molecules, which are based upon a common core nucleotide sequence. We show that neither BLM nor WRN is capable of unwinding duplex DNA from a blunt-ended terminus or from an internal nick. However, both enzymes efficiently unwind the same blunt-ended duplex containing a centrally located 12 nt single-stranded 'bubble', as well as a synthetic X-structure (a model for the Holliday junction recombination intermediate) in which each 'arm' of the 4-way junction is blunt-ended. Surprisingly, a 3'-tailed duplex, a standard substrate for 3'-->5' helicases, is unwound much less efficiently by BLM and WRN than are the bubble and X-structure substrates. These data show conclusively that a single-stranded 3'-tail is not a structural requirement for unwinding of standard B-form DNA by these helicases. BLM and WRN also both unwind a variety of different forms of G-quadruplex DNA, a structure that can form at guanine-rich sequences present at several genomic loci. Our data indicate that BLM and WRN are atypical helicases that are highly DNA structure specific and have similar substrate specificities. We interpret these data in the light of the genomic instability and hyper-recombination characteristics of cells from individuals with Bloom's or Werner's syndrome. PMID- 11433032 TI - Genome-wide detection of alternative splicing in expressed sequences of human genes. AB - We have identified 6201 alternative splice relationships in human genes, through a genome-wide analysis of expressed sequence tags (ESTs). Starting with approximately 2.1 million human mRNA and EST sequences, we mapped expressed sequences onto the draft human genome sequence and only accepted splices that obeyed the standard splice site consensus. A large fraction (47%) of these were observed multiple times, indicating that they comprise a substantial fraction of the mRNA species. The vast majority of the detected alternative forms appear to be novel, and produce highly specific, biologically meaningful control of function in both known and novel human genes, e.g. specific removal of the lysosomal targeting signal from HLA-DM beta chain, replacement of the C-terminal transmembrane domain and cytoplasmic tail in an FC receptor beta chain homolog with a different transmembrane domain and cytoplasmic tail, likely modulating its signal transduction activity. Our data indicate that a large proportion of human genes, probably 42% or more, are alternatively spliced, but that this appears to be observed mainly in certain types of molecules (e.g. cell surface receptors) and systemic functions, particularly the immune system and nervous system. These results provide a comprehensive dataset for understanding the role of alternative splicing in the human genome, accessible at http://www.bioinformatics.ucla.edu/HASDB. PMID- 11433033 TI - Amino acid-base interactions: a three-dimensional analysis of protein-DNA interactions at an atomic level. AB - To assess whether there are universal rules that govern amino acid-base recognition, we investigate hydrogen bonds, van der Waals contacts and water mediated bonds in 129 protein-DNA complex structures. DNA-backbone interactions are the most numerous, providing stability rather than specificity. For base interactions, there are significant base-amino acid type correlations, which can be rationalised by considering the stereochemistry of protein side chains and the base edges exposed in the DNA structure. Nearly two-thirds of the direct read-out of DNA sequences involves complex networks of hydrogen bonds, which enhance specificity. Two-thirds of all protein-DNA interactions comprise van der Waals contacts, compared to about one-sixth each of hydrogen and water-mediated bonds. This highlights the central importance of these contacts for complex formation, which have previously been relegated to a secondary role. Although common, water mediated bonds are usually non-specific, acting as space-fillers at the protein DNA interface. In conclusion, the majority of amino acid-base interactions observed follow general principles that apply across all protein-DNA complexes, although there are individual exceptions. Therefore, we distinguish between interactions whose specificities are 'universal' and 'context-dependent'. An interactive Web-based atlas of side chain-base contacts provides access to the collected data, including analyses and visualisation of the three-dimensional geometry of the interactions. PMID- 11433034 TI - Translesion synthesis by yeast DNA polymerase zeta from templates containing lesions of ultraviolet radiation and acetylaminofluorene. AB - In the yeast Saccharomyces cerevisiae, DNA polymerase zeta (Polzeta) is required in a major lesion bypass pathway. To help understand the role of Polzeta in lesion bypass, we have performed in vitro biochemical analyses of this polymerase in response to several DNA lesions. Purified yeast Polzeta performed limited translesion synthesis opposite a template TT (6-4) photoproduct, incorporating A or T with similar efficiencies (and less frequently G) opposite the 3' T, and predominantly A opposite the 5' T. Purified yeast Polzeta predominantly incorporated a G opposite an acetylaminofluorene (AAF)-adducted guanine. The lesion, however, significantly inhibited subsequent extension. Furthermore, yeast Polzeta catalyzed extension DNA synthesis from primers annealed opposite the AAF guanine and the 3' T of the TT (6-4) photoproduct with varying efficiencies. Extension synthesis was more efficient when A or C was opposite the AAF-guanine, and when G was opposite the 3' T of the TT (6-4) photoproduct. In contrast, the 3' T of a cis-syn TT dimer completely blocked purified yeast Polzeta, whereas the 5' T was readily bypassed. These results support the following dual-function model of Polzeta. First, Polzeta catalyzes nucleotide incorporation opposite AAF guanine and TT (6-4) photoproduct with a limited efficiency. Secondly, more efficient bypass of these lesions may require nucleotide incorporation by other DNA polymerases followed by extension DNA synthesis by Polzeta. PMID- 11433035 TI - SPINE: an integrated tracking database and data mining approach for identifying feasible targets in high-throughput structural proteomics. AB - High-throughput structural proteomics is expected to generate considerable amounts of data on the progress of structure determination for many proteins. For each protein this includes information about cloning, expression, purification, biophysical characterization and structure determination via NMR spectroscopy or X-ray crystallography. It will be essential to develop specifications and ontologies for standardizing this information to make it amenable to retrospective analysis. To this end we created the SPINE database and analysis system for the Northeast Structural Genomics Consortium. SPINE, which is available at bioinfo.mbb.yale.edu/nesg or nesg.org, is specifically designed to enable distributed scientific collaboration via the Internet. It was designed not just as an information repository but as an active vehicle to standardize proteomics data in a form that would enable systematic data mining. The system features an intuitive user interface for interactive retrieval and modification of expression construct data, query forms designed to track global project progress and external links to many other resources. Currently the database contains experimental data on 985 constructs, of which 740 are drawn from Methanobacterium thermoautotrophicum, 123 from Saccharomyces cerevisiae, 93 from Caenorhabditis elegans and the remainder from other organisms. We developed a comprehensive set of data mining features for each protein, including several related to experimental progress (e.g. expression level, solubility and crystallization) and 42 based on the underlying protein sequence (e.g. amino acid composition, secondary structure and occurrence of low complexity regions). We demonstrate in detail the application of a particular machine learning approach, decision trees, to the tasks of predicting a protein's solubility and propensity to crystallize based on sequence features. We are able to extract a number of key rules from our trees, in particular that soluble proteins tend to have significantly more acidic residues and fewer hydrophobic stretches than insoluble ones. One of the characteristics of proteomics data sets, currently and in the foreseeable future, is their intermediate size ( approximately 500-5000 data points). This creates a number of issues in relation to error estimation. Initially we estimate the overall error in our trees based on standard cross validation. However, this leaves out a significant fraction of the data in model construction and does not give error estimates on individual rules. Therefore, we present alternative methods to estimate the error in particular rules. PMID- 11433036 TI - DNA repair and sequence context affect (1)O(2)-induced mutagenesis in bacteria. AB - Electronic excited molecular oxygen (singlet oxygen, (1)O(2)) is known to damage DNA, yielding mutations. In this work, the mutagenicity induced by (1)O(2) in a defined sequence of DNA was investigated after replication in Escherichia coli mutants deficient for nucleotide and base excision DNA repair pathways. For this purpose a plasmid containing a (1)O(2)-damaged 14 base oligonucleotide was introduced into E.coli by transfection and mutations were screened by hybridization with an oligonucleotide with the original sequence. Mutagenesis was observed in all strains tested, but it was especially high in the BH20 (fpg), AYM57 (fpg mutY) and AYM84 (fpg mutY uvrC) strains. The frequency of mutants in the fpg mutY strain was higher than in the triple mutant fpg mutY uvrC, suggesting that activity of the UvrABC excinuclease can favor the mutagenesis of these lesions. Additionally, most of the mutations were G-->T and G-->C transversions, but this was dependent on the position of the guanine in the sequence and on repair deficiency in the host bacteria. Thus, the kind of repair and the mutagenesis associated with (1)O(2)-induced DNA damage are linked to the context of the damaged sequence. PMID- 11433037 TI - Rapid characterization of DNA oligomers and genotyping of single nucleotide polymorphism using nucleotide-specific mass tags. AB - Using currently available MS-based methods, accurate mass measurements are essential for the characterization of DNA oligomers. However, there is a lack of specificity in mass peaks when the characterization of individual DNA species in a mass spectrum is dependent solely upon the mass-to-charge ratio (m/z). Here, we utilize nucleotide-specific tagging with stable isotopes to provide internal signatures that quantitatively display the nucleotide content of oligomer peaks in MS spectra. The characteristic mass-split patterns induced by the partially (13)C/(15)N-enriched dNTPs in DNA oligomers indicate the number of labeled precursors and in turn the base substitution in each mass peak, and provide for efficient SNP detection. Signals in mass spectra not only reflect the masses of particular DNA oligomers, but also their specific composition of particular nucleotides. The measurements of mass tags are relative in the mass-split pattern and, hence, the accuracy of the determination of nucleotide substitution is indirectly increased. For high sample throughput, (13)C/(15)N-labeled sequences of interest have been generated, excised in solution and purified for MS analysis in a single-tube format. This method can substantially improve the specificity, accuracy and efficiency of mass spectrometry in the characterization of DNA oligomers and genetic variations. PMID- 11433038 TI - Exciton interaction in molecular beacons: a sensitive sensor for short range modifications of the nucleic acid structure. AB - Molecular beacons are hairpin-shaped, single-stranded oligonucleotides constituting sensitive fluorescent DNA probes widely used to report the presence of specific nucleic acids. In its closed form the stem of the hairpin holds the fluorophore covalently attached to one end, close to the quencher, which is covalently attached to the other end. Here we report that in the closed form the fluorophore and the quencher form a ground state intramolecular heterodimer whose spectral properties can be described by exciton theory. Formation of the heterodimers was found to be poorly sensitive to the stem sequence, the respective positions of the dyes and the nature of the nucleic acid (DNA or RNA). The heterodimer allows strong coupling between the transition dipoles of the two chromophores, leading to dramatic changes in the absorption spectrum that are not compatible with a Forster-type fluorescence resonance energy transfer (FRET) mechanism. The excitonic heterodimer and its associated absorption spectrum are extremely sensitive to the orientation of and distance between the dyes. Accordingly, the application of molecular beacons can be extended to monitoring short range modifications of the stem structure. Moreover, the excitonic interaction was also found to operate for doubly end-labeled duplexes. PMID- 11433039 TI - yMGV: a database for visualization and data mining of published genome-wide yeast expression data. AB - The yeast Microarray Global Viewer (yMGV) is an on-line database providing a synthetic view of the transcriptional expression profiles of Saccharomyces cerevisiae genes in most of the published expression datasets. yMGV displays a one-screen graphical representation of gene expression variations for each published genome-wide experiment, allowing quick retrieval of experimental conditions affecting expression of this gene. yMGV also provides tools to isolate groups of genes sharing similar transcription profiles in a defined subset of experiments. Additionally, yMGV furnishes a set of statistical tools for critical assessment of published data. We therefore believe that yMGV is an efficient tool that affords a quick and comprehensive overview of microarray data and generates new gene classifications. As of 20 March 2001 the yMGV database contains 6 000 000 measurements, representing genome-wide expression comparisons of 932 experiments from 39 microarray publications. The yMGV interface is available at http://transcriptome.ens.fr/ymgv/. PMID- 11433040 TI - The mapping of nucleosomes and regulatory protein binding sites at the Saccharomyces cerevisiae MFA2 gene: a high resolution approach. AB - We have developed an end-labelling approach to map the positions of nucleosomes and protein binding sites at nucleotide resolution by footprinting micrococcal nuclease (MNase)-sensitive sites. Using this approach we determined that the MFA2 gene and its upstream control regions have four positioned nucleosomes when transcription is repressed in mating type alpha cells and that the nucleosomes lose their positioning when the gene became transcriptionally active in mating type a cells. We also detected MNase-hypersensitive sites in the alpha2 operator region of MFA2 in alpha cells but not in a cells. These probably result from the change in the local DNA conformation due to protein(s) binding in this region that governs MFA2 transcription. PMID- 11433041 TI - Bisulfite genomic sequencing: systematic investigation of critical experimental parameters. AB - Bisulfite genomic sequencing is the method of choice for the generation of methylation maps with single-base resolution. The method is based on the selective deamination of cytosine to uracil by treatment with bisulfite and the sequencing of subsequently generated PCR products. In contrast to cytosine, 5 methylcytosine does not react with bisulfite and can therefore be distinguished. In order to investigate the potential for optimization of the method and to determine the critical experimental parameters, we determined the influence of incubation time and incubation temperature on the deamination efficiency and measured the degree of DNA degradation during the bisulfite treatment. We found that maximum conversion rates of cytosine occurred at 55 degrees C (4-18 h) and 95 degrees C (1 h). Under these conditions at least 84-96% of the DNA is degraded. To study the impact of primer selection, homologous DNA templates were constructed possessing cytosine-containing and cytosine-free primer binding sites, respectively. The recognition rates for cytosine (>/=97%) and 5 methylcytosine (>/=94%) were found to be identical for both templates. PMID- 11433042 TI - Reproducible and inexpensive probe preparation for oligonucleotide arrays. AB - We present a new protocol for the preparation of nucleic acids for microarray hybridization. DNA is fragmented quantitatively and reproducibly by using a hydroxyl radical-based reaction, which is initiated by hydrogen peroxide, iron(II)-EDTA and ascorbic acid. Following fragmentation, the nucleic acid fragments are densely biotinylated using a biotinylated psoralen analog plus UVA light and hybridized on microarrays. This non-enzymatic protocol circumvents several practical difficulties associated with DNA preparation for microarrays: the lack of reproducible fragmentation patterns associated with enzymatic methods; the large amount of labeled nucleic acids required by some array designs, which is often combined with a limited amount of starting material; and the high cost associated with currently used biotinylation methods. The method is applicable to any form of nucleic acid, but is particularly useful when applying double-stranded DNA on oligonucleotide arrays. Validation of this protocol is demonstrated by hybridizing PCR products with oligonucleotide-coated microspheres and PCR amplified cDNA with Affymetrix Cancer GeneChip microarrays. PMID- 11433043 TI - Novel display of knotted DNA molecules by two-dimensional gel electrophoresis. AB - We describe a two-dimensional agarose gel electrophoresis procedure that improves the resolution of knotted DNA molecules. The first gel dimension is run at low voltage, and DNA knots migrate according to their compactness. The second gel dimension is run at high voltage, and DNA knots migrate according to other physical parameters such as shape and flexibility. In comparison with one dimensional gel electrophoresis, this procedure segregates the knotted DNA molecules from other unknotted forms of DNA, and partially resolves populations of knots that have the same number of crossings. The two-dimensional display may allow quantitative and qualitative characterization of different types of DNA knots simply by gel velocity. PMID- 11433044 TI - Quantification of splice variants using real-time PCR. AB - A reliable and robust method for measuring the expression of alternatively spliced transcripts is an important step in investigating the significance of each variant. So far, accurate quantification of splice variants has been laborious and difficult due to the intrinsic limitations of conventional methods. The many advantages of real-time PCR have made this technique attractive to study its application in quantification of splice isoforms. We use skipping of exon 37 in the NF1 gene as a model to compare and evaluate the different strategies for quantitating splice variants using real-time PCR. An overview of three different possibilities for detecting alternative transcripts is given. We propose the use of a boundary-spanning primer to quantify isoforms that differ greatly in abundance. We describe here a novel method for creating a reliable standard curve using one plasmid containing both alternative transcripts. In addition, we validate the use of an absolute standard curve based on a dilution series of fluorometrically quantified PCR products. PMID- 11433046 TI - Reducing antibiotic use in children: a randomized trial in 12 practices. AB - OBJECTIVE: To test whether an educational outreach intervention for families and physicians, based on the Centers for Disease Control and Prevention (CDC) principles of judicious antibiotic use, decreases antimicrobial drug prescribing for children younger than 6 years old. Setting. Twelve practices affiliated with 2 managed care organizations (MCOs) in eastern Massachusetts and northwest Washington State. Patients. All enrolled children younger than 6 years old. METHODS: Practices stratified by MCO and size were randomized to intervention or control groups. The intervention included 2 meetings of the practice with a physician peer leader, using CDC-endorsed summaries of judicious prescribing recommendations; feedback on previous prescribing rates were also provided. Parents were mailed a CDC brochure on antibiotic use, and supporting materials were displayed in waiting rooms. Automated enrollment, ambulatory visit, and pharmacy claims were used to determine rates of antibiotic courses dispensed (antibiotics/person-year) during baseline (1996-1997) and intervention (1997 1998) years. The primary analysis (for children 3 to <36 months and 36 to <72 months) assessed the impact of the intervention among children during the intervention year, controlling for covariates including patient age and baseline prescription rate. Confirmatory analyses at the practice level were also performed. RESULTS: The practices cared for 14 468 and 13 460 children in the 2 study years, respectively; 8815 children contributed data in both years. Sixty two percent of antibiotic courses were dispensed for otitis media, 6.5% for pharyngitis, 6.3% for sinusitis, and 9.2% for colds and bronchitis. Antibiotic dispensing for children 3 to <36 months old decreased 0.41 antibiotics per person year (18.6%) in intervention compared with 0.33 (11.5%) in control practices. Among children 36 to <72 months old, the rate decreased by 0.21 antibiotics per person-year (15%) in intervention and 0.17 (9.8%) in control practices. Multivariate analysis showed an adjusted intervention effect of 16% in the younger and 12% in the older age groups. The direction and approximate magnitude of effect were confirmed in practice-level analyses. CONCLUSIONS: A limited simultaneous educational outreach intervention for parents and providers reduced antibiotic use among children in primary care practices, even in the setting of substantial secular trends toward decreased prescribing. Future efforts to promote judicious prescribing should continue to build on growing public awareness of antibiotic overuse. PMID- 11433045 TI - Minisequencing on oligonucleotide microarrays: comparison of immobilisation chemistries. AB - In the microarray format of the minisequencing method multiple oligonucleotide primers immobilised on a glass surface are extended with fluorescent ddNTPs using a DNA polymerase. The method is a promising tool for large-scale single nucleotide polymorphism (SNP) detection. We have compared eight chemical methods for covalent immobilisation of the oligonucleotide primers on glass surfaces. We included both commercially available, activated slides and slides that were modified by ourselves. In the comparison the differently derivatised glass slides were evaluated with respect to background fluorescence, efficiency of attaching oligonucleotides and performance of the primer arrays in minisequencing reactions. We found that there are significant differences in background fluorescence levels among the different coatings, and that the attachment efficiency, which was measured indirectly using extension by terminal transferase, varied largely depending on which immobilisation strategy was used. We also found that the attachment chemistry affects the genotyping accuracy, when minisequencing on microarrays is used as the genotyping method. The best genotyping results were observed using mercaptosilane-coated slides attaching disulfide-modified oligonucleotides. PMID- 11433047 TI - Diagnostic accuracy of screening electrocardiograms in long QT syndrome I. AB - OBJECTIVE: Inherited long QT syndrome (LQTS) may present with syncope, seizures, and/or sudden death as a result of ventricular tachyarrhythmias. Identification of family members who are at risk because they harbor the genetic substrate for LQTS is critical. Presently, such identification relies on the 12-lead electrocardiogram (ECG). The purpose of this study was to evaluate the efficacy of the automated ECG as a screening tool for LQTS. METHOD: Molecular testing of a proband and 22 additional family members for the KVLQT1 mutation and symptomatic status facilitated the classification of each family member into the following patient groups: noncarriers (13), asymptomatic carriers (5), and symptomatic carriers (5). Each individual had a standard 12-lead ECG from which the computer and manual (lead II) corrected QT interval were determined. In addition, we determined the accuracy of the computer ECG diagnostic interpretation for each patient group. RESULTS: With the use of a corrected QT interval of >/=460 ms as a diagnostic cutoff, the positive and negative predictive values for identifying at risk individuals were 100%. Despite this, the computer-generated ECG diagnostic interpretation erroneously classified 6 of 23 family members. Moreover, half of the family members, proved to have the ion channel defect, received the diagnostic interpretation "normal ECG." CONCLUSION: Reliance on the computer generated ECG diagnostic interpretation alone will fail to identify many at-risk family members. It is suggested that all first-degree relatives of an identified LQTS proband have a 12-lead ECG that is reviewed independently by a physician who is familiar with LQTS in an effort to improve screening for this potentially lethal syndrome.electrocardiogram, long QT syndrome, QT interval, sudden death. PMID- 11433048 TI - A prospective randomized, controlled trial comparing synchronized nasal intermittent positive pressure ventilation versus nasal continuous positive airway pressure as modes of extubation. AB - OBJECTIVE: To determine whether synchronized nasal intermittent positive pressure ventilation (SNIPPV) would decrease extubation failure compared with nasal continuous positive airway pressure (NCPAP) in preterm infants being ventilated for respiratory distress syndrome (RDS). METHODS: Infants who were /=0.5 mL/kg/cm H(2)O and expiratory airway resistance of 70 cm H(2)O/L/s), successful extubation was seen in 93% (27 of 29) in the SNIPPV group and 60% (15 of 25) in the NCPAP group. When weight was controlled for at the time of extubation, the odds of success in the SNIPPV group were 21.1 times higher (95% confidence interval: 3.4, 130.1) than that of the NCPAP group. CONCLUSIONS: SNIPPV is more effective than NCPAP in weaning infants with RDS from the ventilator. PFT may be useful in predicting successful extubation. PMID- 11433049 TI - A randomized trial to reduce passive smoke exposure in low-income households with young children. AB - OBJECTIVE: Passive smoke exposure among children is widespread in the United States; estimates suggest that almost 40% of children who are younger than 5 years live with a smoker. Few randomized studies of passive smoke exposure reduction among children have been conducted, and the impact of interventions that have been evaluated has been limited. The objective of this study was to determine whether a motivational intervention for smoking parents of young children will lead to reduced household passive smoke exposure. METHODS: Project KISS (Keeping Infants Safe From Smoke), a theory-driven exposure reduction intervention targeting low-income families with young children, was a randomized controlled study in which participants-smoking parents/caregivers (N = 291) who had children who were younger than 3 years and who were recruited through primary care settings-were randomly assigned to either the motivational intervention (MI) or a self-help (SH) comparison condition was used. Follow-up assessments were conducted at 3 and 6 months. The MI condition consisted of a 30- to 45-minute motivational interviewing session at the participant's home with a trained health educator and 4 follow-up telephone counseling calls. Feedback from baseline household air nicotine assessments and assessment of the participant's carbon monoxide level was provided as part of the intervention. Participants in the SH group received a copy of the smoking cessation manual, the passive smoke reduction tip sheet, and the resource guide in the mail. Household nicotine levels were measured by a passive diffusion monitor. RESULTS: The 6-month nicotine levels were significantly lower in MI households. Repeated measures analysis of variance across baseline, 3-month, and 6-month time points showed a significant time-by-treatment interaction, whereby nicotine levels for the MI group decreased significantly and nicotine levels for the SH group increased but were not significantly different from baseline. CONCLUSIONS: This study targeted a large sample of racially and ethnically diverse low-income families, in whom both exposure and disease burden is likely to be significant. This is the first study to our knowledge that has been effective in reducing objective measures of passive smoke exposure in households with healthy children. These findings have important implications for pediatric health care providers, who play an important role in working with parents to protect children's health. Providers can help parents work toward reducing household passive smoke exposure using motivational strategies and providing a menu of approaches regardless of whether the parents are ready to quit. PMID- 11433050 TI - A single dose of Sn-mesoporphyrin prevents development of severe hyperbilirubinemia in glucose-6-phosphate dehydrogenase-deficient newborns. AB - OBJECTIVES: Severe neonatal jaundice is a common clinical manifestation of glucose-6-phosphate dehydrogenase (G-6-PD) deficiency and the most difficult to manage; kernicterus is not an uncommon outcome. We assessed in healthy, direct Coombs test-negative Greek newborns of >/=38 weeks' gestational age 1) the current burden of G-6-PD deficiency-associated severe jaundice, and 2) the efficacy of preventive use of Sn-mesoporphyrin (SnMP), a potent inhibitor of heme oxygenase activity and thus of bilirubin production, in ameliorating jaundice in G-6-PD-deficient neonates. METHODS: The studies were conducted at Metera Maternity Hospital in Athens, Greece. Enrolled newborns had the plasma bilirubin concentration (PBC) determined in cord blood and daily thereafter until a declining level was obtained and the case was closed. Intervention with phototherapy was dictated at exact, age-specific PBC levels. In our initial study, we enrolled consecutive mature healthy G-6-PD-deficient newborns as well as a threefold excess of G-6-PD-normal neonates born at approximately the same time (control group). For the SnMP trial, G-6-PD-deficient neonates were administered SnMP as a single intramuscular dose of 6 micromol/kg birth weight within 24 +/- 12 hours of age. RESULTS: SnMP was administered at 26.7 +/- 6.1 hours of age to 172 G-6-PD-deficient newborns (group A); 168 G-6-PD-normal (group B) and 58 G-6-PD-deficient (group C) newborns who were enrolled earlier provided the comparison groups. Except for the expected excess of males in the G-6-PD deficient groups (A and C), there were no differences in the demographic characteristics among the 3 groups. The incremental changes in PBC from cord blood to 24 hours of age also were similar (group A: 4.13 +/- 1.32 mg/dL; group B: 4.05 +/- 1.34 mg/dL; group C: 4.39 +/- 1.07 mg/dL), but there were significant differences in the next period, 24 to 48 hours of age (group A: 0.63 +/- 1.44 mg/dL; group B: 1.69 +/- 1.5 mg/dL; group C: 2.45 +/- 1.72 mg/dL). Peak PBC was significantly different (group A: 7.81 +/- 3.04 mg/dL; group B: 8.68 +/- 3.1 mg/dL; group C: 11.24 +/- 3.76 mg/dL) as was the age at which peak PBC was recorded (group A: 56 +/- 29 hours of age; group B: 69 +/- 26 hours of age; group C: 83 +/- 29 hours of age). These differences in favor of group A were observed despite the fact that phototherapy was used in 15% of the newborns in group B and 31% of those in group C, whereas none of those treated with SnMP required phototherapy. Finally, in one female, who was heterozygous for G-6-PD deficiency, in group C phototherapy failed and 2 exchange transfusions were performed. CONCLUSIONS: In comparison with normal neonates, G-6-PD-deficient neonates experienced a twofold increase in the prevalence of significant hyperbilirubinemia requiring phototherapy. A single dose of SnMP administered in the 1st day of life to the G-6-PD-deficient newborns shifted the peak PBC distribution to the left (lower values) even in relation to normal neonates and entirely eliminated the need for phototherapy. Interdiction of bilirubin production by use of a heme oxygenase inhibitor such as SnMP represents a simple and highly effective means for the preventive management of jaundice in G-6-PD deficient newborns. PMID- 11433051 TI - Prediction of hyperbilirubinemia in near-term and term infants. AB - OBJECTIVE: The purpose of this study was to determine whether end-tidal carbon monoxide (CO) corrected for ambient CO (ETCOc), as a single measurement or in combination with serum total bilirubin (STB) measurements, can predict the development of hyperbilirubinemia during the first 7 days of life. METHODS: From 9 multinational clinical sites, 1370 neonates completed this cohort study from February 20, 1998, through February 22, 1999. Measurements of both ETCOc and STB were performed at 30 +/- 6 hours of life; STB also was measured at 96 +/- 12 hours and subsequently following a flow diagram based on a table of hours of age specific STB. An infant was defined as hyperbilirubinemic if the hours of age specific STB was greater than or equal to the 95th percentile as defined by the table at any time during the study. RESULTS: A total of 120 (8.8%) of the enrolled infants became hyperbilirubinemic. Mean STB in breastfed infants was 8.92 +/- 4.37 mg/dL at 96 hours versus 7.63 +/- 3.58 mg/dL in those fed formula only. The mean ETCOc at 30 +/- 6 hours for the total population was 1.48 +/- 0.49 ppm, whereas those of nonhyperbilirubinemic and hyperbilirubinemic infants were 1.45 +/- 0.47 ppm and 1.81 +/- 0.59 ppm, respectively. Seventy-six percent (92 of 120) of hyperbilirubinemic infants had ETCOc greater than the population mean. An ETCOc greater than the population mean at 30 +/- 6 hours yielded a 13.0% positive predictive value (PPV) and a 95.8% negative predictive value (NPV) for STB >/=95th percentile. When infants with STB >95th percentile at <36 hours of age were excluded, the STB at 30 +/- 6 hours yielded a 16.7% PPV and a 98.1% NPV for STB >75th percentile. The combination of these 2 measurements at 30 +/- 6 hours (either ETCOc more than the population mean or STB >75th percentile) had a 6.4% PPV with a 99.0% NPV. Conclusions. This prospective cohort study supports previous observations that measuring STB before discharge may provide some assistance in predicting an infant's risk for developing hyperbilirubinemia. The addition of an ETCOc measurement provides insight into the processes that contribute to the condition but does not materially improve the predictive ability of an hours of age-specific STB in this study population. The combination of STB and ETCOc as early as 30 +/- 6 hours may identify infants with increased bilirubin production (eg, hemolysis) or decreased elimination (conjugation defects) as well as infants who require early follow-up after discharge for jaundice or other clinical problems such as late anemia. Depending on the incidence of hyperbilirubinemia within an institution, the criteria for decision making should vary according to its unique population. PMID- 11433052 TI - Estimated prevalence of noise-induced hearing threshold shifts among children 6 to 19 years of age: the Third National Health and Nutrition Examination Survey, 1988-1994, United States. AB - OBJECTIVE: This analysis estimates the first nationally representative prevalence of noise-induced hearing threshold shifts (NITS) among US children. Historically, NITS has not been considered a common cause of childhood hearing problems. Among children, NITS can be a progressive problem with continued exposure to excessive noise, which can lead to high-frequency sound discrimination difficulties (eg, speech consonants and whistles). METHODS: The Third National Health and Nutrition Examination Survey (NHANES III) was conducted from 1988 to 1994. NHANES III is a national population-based cross-sectional survey with a household interview, audiometric testing at 0.5 to 8 kHz, and compliance testing. A total of 5249 children aged 6 to 19 years completed audiometry and compliance testing for both ears in NHANES III. The criteria used to assess NITS included audiometry indicating a noise notch in at least 1 ear. RESULTS: Of US children 6 to 19 years old, 12.5% (approximately 5.2 million) are estimated to have NITS in 1 or both ears. In the majority of the children meeting NITS criteria, only 1 ear and only 1 frequency are affected. In this analysis, all children identified with NITS passed compliance testing, which essentially rules out middle ear disorders such as conductive hearing loss. The prevalence estimate of NITS differed by sociodemographics, including age and sex. CONCLUSIONS: These findings suggest that children are being exposed to excessive amounts of hazardous levels of noise, and children's hearing is vulnerable to these exposures. These data support the need for research on appropriate hearing conservation methods and for NITS screening programs among school-aged children. Public health interventions such as education, training, audiometric testing, exposure assessment, hearing protection, and noise control when feasible are all components of occupational hearing conservation that could be adapted to children's needs with children specific research. PMID- 11433053 TI - Food insufficiency and American school-aged children's cognitive, academic, and psychosocial development. AB - OBJECTIVE: This study investigates associations between food insufficiency and cognitive, academic, and psychosocial outcomes for US children and teenagers ages 6 to 11 and 12 to 16 years. METHODS: Data from the Third National Health and Nutrition Examination Survey (NHANES III) were analyzed. Children were classified as food-insufficient if the family respondent reported that his or her family sometimes or often did not get enough food to eat. Regression analyses were conducted to test for associations between food insufficiency and cognitive, academic, and psychosocial measures in general and then within lower-risk and higher-risk groups. Regression coefficients and odds ratios for food insufficiency are reported, adjusted for poverty status and other potential confounding factors. RESULTS: After adjusting for confounding variables, 6- to 11 year-old food-insufficient children had significantly lower arithmetic scores and were more likely to have repeated a grade, have seen a psychologist, and have had difficulty getting along with other children. Food-insufficient teenagers were more likely to have seen a psychologist, have been suspended from school, and have had difficulty getting along with other children. Further analyses divided children into lower-risk and higher-risk groups. The associations between food insufficiency and children's outcomes varied by level of risk. CONCLUSIONS: The results demonstrate that negative academic and psychosocial outcomes are associated with family-level food insufficiency and provide support for public health efforts to increase the food security of American families. PMID- 11433054 TI - Intussusception, rotavirus diarrhea, and rotavirus vaccine use among children in New York state. AB - OBJECTIVE: To describe epidemiologic features of intussusception and rotavirus diarrhea in New York, to examine the baseline incidence and trends over time, and to ascertain whether an excess of cases occurred in the 9 months of vaccination with the newly licensed rotavirus vaccine. METHODS: Hospital discharge data from 1989 through 1998 were reviewed for children (<1 year old) whose primary or secondary diagnosis was coded as intussusception or rotavirus diarrhea. Characteristics of patients admitted for intussusception and rotavirus diarrhea were compared, and trends over time were examined. For a subset of patients, medical records and vaccine histories for intussusception hospitalizations from October 1998 through June 1999 were analyzed. The number of intussusception cases attributable to rotavirus vaccine was calculated based on the penetration of the vaccine (21%) and a range of excess risks of intussusception among vaccinated children as estimated by the National Immunization Program (NIP). RESULTS: From 1989 through 1998, 1450 intussusception-associated hospitalizations were reported in children <1 year old (average annual incidence 5.4/10 000). Among these children, 47% were treated medically and 53% had surgery, with 9% needing surgical resection. The incidence of intussusception declined over time from 6.1 per 10 000 in 1989 to 3.9 per 10 000 in 1998. Intussusception hospitalizations occurred throughout the year, whereas rotavirus-associated hospitalizations peaked from February to April. Of 20 patients with intussusception whose hospitalization charts were reviewed, 5 had received rotavirus vaccine. All 5 were hospitalized after their first dose of vaccine, were admitted before 7 months of age, were white, and had private insurance. A total of 81 cases of intussusception occurred during the 9-month period of rotavirus vaccination, compared with 78 during the same period in the prevaccination year. The number of excess intussusception cases observed (n = 3) was lower than expected using the NIP estimate of excess risk (1.8) among rotavirus vaccinated children (n = 12) but not significantly different from the risks identified in the NIP cohort studies (1 in 12 000). CONCLUSION: Our data suggest that in New York the rate of intussusception has declined, and approximately 1 child in 2600 develops intussusception before 1 year of age. The different seasonality between intussusception and rotavirus-related hospitalizations suggests that if any causal association exists, it must be small. Unlike other studies, analysis of New York hospitalized discharge data failed to show an appreciable increase in the incidence of intussusception after introduction of the rotavirus vaccine. PMID- 11433055 TI - Allopurinol neurocardiac protection trial in infants undergoing heart surgery using deep hypothermic circulatory arrest. AB - OBJECTIVE: This pharmacologic protection trial was conducted to test the hypothesis that allopurinol, a scavenger and inhibitor of oxygen free radical production, could reduce death, seizures, coma, and cardiac events in infants who underwent heart surgery using deep hypothermic circulatory arrest (DHCA). DESIGN: This was a single center, randomized, placebo-controlled, blinded trial of allopurinol in infant heart surgery using DHCA. Enrolled infants were stratified as having hypoplastic left heart syndrome (HLHS) and all other forms of congenital heart disease (non-HLHS). Drug was administered before, during, and after surgery. Adverse events and the clinical efficacy endpoints death, seizures, coma, and cardiac events were monitored until infants were discharged from the intensive care unit or 6 weeks, whichever came first. RESULTS: Between July 1992 and September 1997, 350 infants were enrolled and 348 subsequently randomized. A total of 318 infants (131 HLHS and 187 non-HLHS) underwent heart surgery using DHCA. There was a nonsignificant treatment effect for the primary efficacy endpoint analysis (death, seizures, and coma), which was consistent over the 2 strata. The addition of cardiac events to the primary endpoint resulted in a lack of consistency of treatment effect over strata, with the allopurinol treatment group experiencing fewer events (38% vs 60%) in the entire HLHS stratum, compared with the non-HLHS stratum (30% vs 27%). In HLHS surgical survivors, 40 of 47 (85%) allopurinol-treated infants did not experience any endpoint event, compared with 27 of 49 (55%) controls. There were fewer seizures only and cardiac-only events in the allopurinol versus placebo groups. Allopurinol did not reduce efficacy endpoint events in non-HLHS infants. Treated and control infants did not differ in adverse events. CONCLUSIONS: Allopurinol provided significant neurocardiac protection in higher-risk HLHS infants who underwent cardiac surgery using DHCA. No benefits were demonstrated in lower risk, non-HLHS infants, and no significant adverse events were associated with allopurinol treatment.congenital heart defects, hypoplastic left heart syndrome, induced hypothermia, ischemia-reperfusion injury, neuroprotective agents, allopurinol, xanthine oxidase, free radicals, seizures, coma. PMID- 11433056 TI - Utilization and cost of health care services for children with attention deficit/hyperactivity disorder. AB - BACKGROUND: Despite an increasing prevalence of diagnosed attention deficit/hyperactivity disorder (ADHD) among children, the impact of ADHD on utilization and costs of health care services is largely unknown. OBJECTIVE: To examine differences in health care utilization and costs between children with and without ADHD. DESIGN: Retrospective matched cohort study conducted from January 1 to December 31, 1997. Setting. Health maintenance organization in western Washington State. PARTICIPANTS: Children aged 3 through 17 years who were continuously enrolled in the health maintenance organization and used services during 1997 were eligible. Children were identified with ADHD if they had a diagnosis of ADHD or a prescription for a stimulant medication using automated patient files. Children without ADHD were randomly selected as controls and matched 4:1 to children with ADHD on age and sex. OUTCOME MEASURE: Utilization and costs of specific categories of health care services. Results. A total of 2992 children (5.2%) were identified with ADHD. Children with ADHD incurred significantly greater per capita total costs ($1465 vs $690) than children without ADHD. Children with ADHD had 9.9 times more outpatient mental health visits (1.35/year vs 0.14/year), 3.4 times more pharmacy fills (11.25/year vs 3.30/year), and 1.6 times more primary care visits (3.84/year vs 2.36/year) than children without ADHD. The adjusted incremental costs were estimated to be $375 (95% confidence interval: $336-$416) for children with ADHD alone and $812 (95% confidence interval: $671-$973) for children with ADHD plus coexisting mental health disorders. CONCLUSIONS: Children with ADHD use significantly more health care resources and incur significantly higher costs than children without ADHD. Coexisting mental health disorders substantially increase the cost of treating ADHD. Resource allocation decisions should consider the contributions of primary care, outpatient mental health, and pharmacy costs to the overall costs of care for children with ADHD. PMID- 11433057 TI - Characteristics and financial performance of a pediatric faculty inpatient attending service: a resource-based relative value scale analysis. AB - OBJECTIVES: In many children's hospitals, inpatient attending physician services are provided by academic faculty who function as part-time inpatient specialists or hospitalists. Although some have claimed that hospitalist care can reduce length of stay and total hospital resource use and expenses, there are few benchmarks or data regarding physician productivity or the characteristics and financial performance of these programs. The resource-based relative value scale (RBRVS) is a valuable tool for developing national benchmarks and comparing the financial performance of inpatient programs at varying daily census and reimbursement levels. The objectives of this study were to 1) describe physician productivity on an inpatient service as measured by total relative value units (TRVUs) and professional charges, 2) determine whether inpatient collections were adequate to support faculty salaries for the time spent attending, and 3) develop a model to evaluate financial performance of inpatient programs at varying census and TRVU reimbursement levels. METHODS: A retrospective review of hospital discharge and faculty practice billing data between June 1997 and July 1998 was conducted in a general medical service in a regional, 208-bed, university affiliated children's hospital in the Pacific Northwest. RESULTS: Of 4113 patients who were admitted to the children's hospital general medical service during a 12-month period, faculty part-time hospitalists (N = 28) served as the attending physician for 1738 (42%). On an annual basis, faculty attended for an average of 29.1 days (median: 21.0; range: 7.0-97.0), with an average daily patient census (ADC) of 7.2 (median: 6.5; range: 2.8-12.0). Inpatient attendings billed for 1738 initial visits and 3957 subsequent visits. Total physician productivity for the inpatient attending group during 1 year included 12 085 TRVUs and gross professional charges of $777 743. The average payment, or conversion factor (CF), was $24.46/TRVU (71% of Medicare CF). The cash collection rate was 38%, reflecting a payor mix that included 54% Medicaid, 28% commercial payors, 12% health maintenance organization, and 6% other payors. On a weekly basis, physicians generated an average of 109 TRVUs and collected $2665 in cash. The average salary cost per RVU was $23.40, and weekly faculty salary and benefit expenses were $2550. After operating expenses and academic taxes totaling 24% were deducted ($5.87/TRVU), RBRVS-based payments and cash collections covered 79% of average faculty weekly salaries. Financial modeling showed that either an average CF of $31/TRVU or an ADC of 9 patients per day on the inpatient service would be required to generate sufficient revenue to support physician salaries and operating expenses. CONCLUSIONS: For a faculty inpatient attending service in a children's hospital with an ADC of 7, a $24.46 RBRVS-based CF payment is inadequate to support faculty salaries and operating expenses for the time spent attending. Inpatient services in similar payor environments with comparable expenses and staffed by faculty who care for fewer than 9 patients per day will not cover typical faculty salary costs and operating expenses. PMID- 11433058 TI - Where children drown, United States, 1995. AB - CONTEXT: Drowning is the second leading cause of unintentional injury death among children ages 1 to 19 years. Details about the specific site of submersion are important for implementation of primary prevention efforts, but are not routinely available from national data. OBJECTIVES: To provide national data about the specific sites of drowning among US children and to examine site-specific drowning rates by age, race, and gender. SETTING: United States, 1995. DESIGN: Information was abstracted from death certificates for unintentional drownings among children <20 years of age. The site of drowning was coded from free text on death certificates. Proportional distributions of the site of drowning by age, region, gender, and race were examined. Site-specific drowning rates were calculated by age, race, and gender. Denominators were based on US Census Bureau, June 1995 estimates of the US population. RESULTS: For 1995, death certificates were obtained for 1420 unintentional drownings among children <20 years of age, 98% of the number of drowning deaths reported by the National Center for Health Statistics for the same year. Site of drowning was specified on 1308 (92%) death certificates. Fifty-five percent of infant drownings were in bathtubs. Among children between the ages of 1 to 4 years, 56% of drownings were in artificial pools and 26% were in other bodies of freshwater. Among older children, 63% of drownings were in natural bodies of freshwater. Site-specific drowning rates varied by race. Importantly, after the age of 5 years, the risk of drowning in a swimming pool was greater among black males compared with white males with rate ratios of 15.1 (95% confidence interval: 6.7-38.5) among 10- to 14-year-olds and 12.8 (95% confidence interval: 6.5-26.9) among 15- to 19-year-olds. CONCLUSION: The variety of sites in which children drown, even within specific age groups, emphasizes the need for a multifaceted approach to prevention. Reductions in the relatively high rates of drowning among black adolescent males will require targeted interventions to prevent swimming pool drownings among this group. PMID- 11433059 TI - Effectiveness of compliance with pediatric preventive care guidelines among Medicaid beneficiaries. AB - OBJECTIVE: Because research has not confirmed a relationship between compliance with health supervision in infancy and improved health outcomes, we examined the association between adherence to prevailing guidelines for periodic health supervision and adverse health outcome indicated by incidence of avoidable hospitalizations. METHODS: This was a historic cohort study of 308 131 children enrolled in Medicaid at birth in California, Georgia, and Michigan in 1990 using Medicaid records linked across 3 years. We used avoidable hospitalizations as indicators of health in a survival analysis. The analysis used variables that represented completeness and timeliness of well-child visits and immunizations using AAP guidelines for health supervision as the gold standard. RESULTS: When the children in this cohort were up-to-date for age on their schedule of well child visits, they were less likely to have an avoidable hospitalization (race, illness, and level of poverty adjusted hazard ratios 0.52 [95% confidence interval (CI): 0.50-0.55] in California, 0.54 [95% CI: 0.50-0.55] in Georgia, and 0.7 [95% CI: 0.69-0.79] in Michigan). Among children who were not up-to-date with well-child visits, a sporadic preventive care visit conferred a mild benefit. Immunizations and race/ethnicity had no consistent relationship with incidence of avoidable hospitalizations. CONCLUSIONS: A series of well-child visits maintained during the first 2 years of life has a positive effect on health outcomes as indicated by a decrease in avoidable hospitalizations among poor and near-poor children, regardless of race, level of poverty, or health status. National efforts to improve the quality of child health services for young children should focus on increasing compliance with periodic preventive care for young children in addition to improving immunization levels. PMID- 11433060 TI - Barriers to domestic violence screening in the pediatric setting. AB - OBJECTIVE: By surveying practitioners in our community, we hoped to determine what pediatricians and family physicians (FPs) perceive as barriers to the American Academy of Pediatrics (AAP) Recommendation on Domestic Violence Screening. BACKGROUND: When screened in the pediatric setting, as many as 40% of mothers will disclose domestic violence (DV) by their partner. Recognizing the profound effects of DV on children, the AAP recently recommended that all practitioners incorporate DV screening as a part of routine anticipatory guidance. Yet, there is little information about whether pediatricians have the training, the time to screen, or understand the magnitude of this problem. DESIGN/METHODS: A 22-question survey about attitudes, training, and current DV screening practice was sent to all general pediatricians and FPs with admitting privileges to Children's Hospital Medical Center in Cincinnati, Ohio. A copy of the AAP recommendation on screening was included. The vast majority of practitioners with an appreciable pediatric practice in the surrounding tri-state area of 1.8 million people have privileges at the institution. RESULTS: After 2 mailings, 310 (57%) of 547 of questionnaires were returned. The majority of practitioners (64%) were unaware of the AAP recommendation, but 51% of practitioners screened at least high-risk families for DV and 49% had identified a case of DV in their practice. Still, only 8.5% routinely screened for DV and 74% had received no specific DV training. Fifty-eight percent of practitioners estimated the incidence of DV to be <5% in their practice. The most commonly perceived barriers to screening were lack of education (61%), office protocol (60%), time (59%), and support staff (55%). FPs were significantly more likely to have DV training (64% vs 21%), more likely to screen at least high-risk women (79% vs 56%), and more likely to have identified a case of DV (92% vs 40%) than pediatricians. FPs were less likely to cite lack of education (46% vs 65%) and lack of time (50% vs 61%) than pediatricians. Physicians licensed in Ohio were less likely to have specific domestic violence training (23% vs 60%) as compared with Kentucky physicians, where domestic violence education is required for licensing. Kentucky physicians were less likely to cite lack of education as barrier to DV screening (20% vs 62%). When comparing the characteristics of those who screen to those who do not, those with DV training were 10.9 times (odds adjusted ratio) more likely to screen. CONCLUSIONS: Practitioners grossly underestimate the incidence of DV in their practices. Lack of education including knowledge of screening recommendations is a barrier to DV screening by pediatricians. Greater efforts are needed to educate pediatricians on DV for the AAP recommendations to be accepted.domestic violence, child abuse, screening, physician attitude. PMID- 11433061 TI - Effects of Medicaid managed care on health care use: infant emergency department and ambulatory services. AB - OBJECTIVE: Many urban children rely on emergency departments (ED) for ambulatory care. The objective of this study was to determine whether enrollment in Medicaid managed care (MMC) alters ED or other ambulatory care compared with fee-for service Medicaid (FFSM). METHODS: A prospective cohort study of infants born between May 1994 and April 1995 with a 6-month follow-up period was conducted in an urban, teaching hospital and surrounding ambulatory settings. A consecutive sample of 644 infants enrolled in MMC or FFSM was studied; 92% of eligible patients were enrolled, and 94% completed follow-up. The main outcome measures were 1) proportion of patients in each group visiting an ED, primary care practitioner (PCP), or specialist; 2) mean number of visits per group; and 3) ED reliance (EDR) defined as the proportion of all ambulatory visits occurring in an ED. RESULTS: Fifty-six percent of MMC and 54% of FFSM patients visited an ED (relative risk: 1.03; 95% confidence interval [CI]: 0.83, 1.27). More MMC patients had a sick visit to their PCP (relative risk: 1.34; 95% CI: 1.03, 1.74); no difference in proportion with well-child or specialty visits was found. Although the mean number of total ambulatory, ED, and specialty visits was the same, MMC patients had fewer well-child and more sick visits to the PCP than FFSM patients (P =.01). EDR was 21% for both groups (P =.95). After adjustment for other factors in multivariate analysis, insurance status remained unassociated with EDR (adjusted odds ratio [OR]: 0.91; 95% CI: 0.56, 1.69). Factors associated with EDR included United States-born mother (OR: 5.34; 95% CI: 1.61, 17.68) and use of a hospital-based primary care physician (OR: 2.00; 95% CI: 1.34, 2.98). Variables that characterized infants who were less likely to be ED reliant included adequate maternal prenatal care (OR: 0.52; 95% CI: 0.34, 0.78) and having a mother who completed high school (OR: 0.67; 95% CI: 0.45, 0.99). CONCLUSIONS: Enrollment in MMC did not alter ED usage patterns when compared with FFSM. Some variation in use of other ambulatory services was detected. PMID- 11433062 TI - School difficulties in 20-year-olds who were born small for gestational age at term in a regional cohort study. AB - OBJECTIVE: To investigate the relation between school difficulties and being born small for gestational age (SGA) at full term in adolescents and young adults. METHODS: A total of 236 full-term singletons who were born SGA (birth weight and/or length below the third percentile) from 1971 through 1978 and 281 full term singletons who were born appropriate for gestational age (AGA; between the 25th and 75th percentiles) from the maternity registry of Haguenau, France. Participants were evaluated at a mean age of 20.6 (+/-2.1) years. The outcomes measured were late entry into secondary school (normal age: 11 years) and failure to take or pass the baccalaureate examination at the end of secondary school (normal age: 18 years). RESULTS: Late entry into secondary school was more frequent for the SGA than the AGA children (odds ratio: 2.3) after adjustment for maternal age and educational level, parental socioeconomic status, family size, and gender. A significantly higher proportion of term SGA adolescents failed to take or pass the baccalaureate examination than AGA adolescents (odds ratio: 1.6). SGA participants with a smaller head circumference entered secondary school late more often than SGA participants with a larger head circumference, but the association was not significant after adjustment. CONCLUSION: Being born SGA at term is associated with poorer school performance at 12 and 18 years. Fetal adaptation to conditions that retard growth during gestation may not be successful in maintaining brain development. PMID- 11433063 TI - Fluctuations in symptoms in human immunodeficiency virus-infected children: the first 10 years of life. AB - OBJECTIVE: To describe patterns of clinical and immunologic progression in children who are vertically infected with the human immunodeficiency virus. METHODS: Children who were born to mothers who were infected with the human immunodeficiency virus in 11 European centers were enrolled at birth in a prospective study and followed according to a standard protocol. At each visit, a clinical and immunologic class was allocated according to guidelines of the Centers for Disease Control and Prevention (CDC). Progression to serious disease and death was assessed, allowing for available and actual antiretroviral therapy (ART). CDC class at each visit was assessed cross-sectionally. RESULTS: More than 15% of infected children will have progressed to category C or death by age 1 year and nearly 50% by 10 years. Just under 20% of children will have evidence of severe immunodeficiency by age 1 and 75% by 10 years. In general, immune status poorly reflected clinical condition. Children who were born after 1994, when the recommendation of earlier initiation of more active therapy was introduced, were significantly less likely to progress than those who were born when treatment was not widely available or was largely confined to zidovudine monotherapy. Estimated progression to CDC class C or death initially was faster in untreated than in treated children, but by 10 years estimated cumulative progression was similar in both groups. Treatment started before class C disease was associated with significantly slower progression. Cross-sectional analysis showed that children largely are symptom-free throughout their lives. After 4 years of age, fewer than 25% of infected children had symptoms at any one time, irrespective of ART received. CONCLUSION: Vertically infected children are without serious symptoms or signs for most of the time. The prognosis has improved with more widespread availability and use of combination ART. These findings have implications for health, education, and other support-service provision. PMID- 11433064 TI - Safety and immunogenicity of a recombinant Borrelia burgdorferi outer surface protein A vaccine against lyme disease in healthy children and adolescents: a randomized controlled trial. AB - OBJECTIVE: A recombinant lipoprotein outer surface protein A (OspA) Lyme disease (LD) vaccine (LYMErix) has been shown to be safe and effective in preventing LD in adults and in adolescents 15 years of age and older. Children are at risk for developing LD. This clinical study was conducted to address the safety and immunogenicity of LD vaccine in children 4 to 18 years of age. METHODS: A randomized, placebo-controlled clinical trial was conducted at 17 investigational sites in Lyme-endemic areas in the United States. Immunogenicity data from this study also were compared with data obtained from the adult efficacy study. A total of 4090 healthy children and adolescents (age range: 4-18; mean age: 10.4 years) were randomized; 4087 were vaccinated, and a subset of 301 children participated in the immunogenicity analysis. Children were randomized to receive either 30 microgram of LD vaccine (N = 3063) or placebo (N = 1024) on a 0, 1, 12 month schedule. Safety assessments evaluated both solicited (local: redness, swelling, and pain; general: fever, headache, fatigue, arthralgia, and rash) and unsolicited adverse events. Serum specimens were collected at month 0 or month 2, and months 6, 12, and 13. RESULTS: Solicited reactogenicity data revealed a higher incidence of local injection site reactions and general symptoms (fever, headache, fatigue, and arthralgia) in vaccine than placebo recipients. The majority of events were limited in duration (mean: 2-3 days) and were mild to moderate in severity. The total IgG anti-OspA geometric mean titer (GMT) in the pediatric vaccine recipients at month 13 was as good as and statistically higher than the GMT in the adult cohort at month 13 (27 485 enzyme-linked immunosorbent assay units [EL.U]/mL vs 8216 EL.U /mL). All of the pediatric vaccine recipients attained a level of antibody concentration >/=1400 EL.U/mL (proposed seroprotective level) compared with 90% of adults attaining levels >/=1400 EL.U/mL in the efficacy trial. CONCLUSIONS: LD vaccine administered on a 0, 1, 12 month schedule generally is well tolerated and immunogenic in children 4 to 18 years of age. The safety profile consists of mild to moderate local injection site reactions and flu-like symptoms of limited duration and did not worsen with subsequent injections. IgG GMT at month 13 was threefold higher than the month 13 GMT obtained in the adult efficacy study. This higher immune response in children should provide protection against LD. PMID- 11433065 TI - Experience with the ketogenic diet in infants. AB - OBJECTIVE: To evaluate the effectiveness, tolerability, and adverse effects of the ketogenic diet in infants with refractory epilepsy. METHODS: A retrospective review of 32 infants who had been treated with the ketogenic diet at a large metropolitan institution. RESULTS: Most infants (71%) were able to maintain strong ketosis. The overall effectiveness of the diet in infants was similar to that reported in the literature for older children; 19.4% became seizure-free, and an additional 35.5% had >50% reduction in seizure frequency. The diet was particularly effective for patients with infantile spasms/myoclonic seizures. There were concomitant reductions in antiepileptic medications. The majority of parents reported improvements in seizure frequency and in their child's behavior and function, particularly with respect to attention/alertness, activity level, and socialization. The diet generally was well-tolerated, and 96.4% maintained appropriate growth parameters. Adverse events, all reversible and occurring in one patient each, included renal stone, gastritis, ulcerative colitis, alteration of mentation, and hyperlipidemia. CONCLUSION: The ketogenic diet should be considered safe and effective treatment for infants with intractable seizures. PMID- 11433066 TI - Outcome at 5 years of age of children 23 to 27 weeks' gestation: refining the prognosis. AB - OBJECTIVES: To determine the changes with postnatal age for survival, with and without major sensorineural disability, to 5 years of age in very preterm infants and to contrast their prognosis with normal birth weight (NBW) control participants. METHODS: A geographically determined cohort study was conducted in Victoria, Australia. Consecutive live births of 23 to 27 weeks' gestational age born during 1991 to 1992 and randomly selected contemporaneous NBW control participants were studied. The main outcome measures were survival and rates of major disability at 5 years of age, determined for those offered intensive care, after day 7, after day 28, and at hospital discharge. RESULTS: Of 401 live births of 23 to 27 weeks' gestation, 225 (56.1%) survived to 5 years of age. The survival rate rose significantly with increasing gestational age at birth in those offered intensive care and by day 7 but not by day 28. The survival rate free of major disability rose significantly with increasing gestational age at all postnatal ages but was not an independent predictive variable by day 28; other adverse events were more important. In the absence of adverse events, the rate of survival free of major disability for very preterm infants who had survived to discharge was 93.2%, similar to the rate of 95.5% for NBW control participants. CONCLUSIONS: The prognosis for very preterm infants changes substantially with postnatal age. Counseling of families should be repeated at intervals, and the advice offered should vary with perinatal events. PMID- 11433067 TI - Childhood neglect and cognitive development in extremely low birth weight infants: a prospective study. AB - OBJECTIVE: To examine the relationship between child maltreatment and cognitive development in extremely low birth weight infants, adjusting for perinatal and parental risk factors. METHODS: A total of 352 infants with birth weight of <1000 g were followed prospectively for 4 years. The data were analyzed with regard to perinatal and parental risk factors and referrals for suspected child maltreatment to government agencies. Perinatal risk factors included birth weight, gestation, gender, periventricular hemorrhage, ventricular dilation, home oxygen requirement, and necrotizing enterocolitis. Parental risk factors included maternal age, race, marital status, education, and hospital insurance status. Cognitive z scores were calculated at 1, 2, and 4 years, and head circumference z scores were calculated at birth, 2 years, and 4 years. RESULTS: Fifteen percent of infants were referred to child protective services for suspected child maltreatment. The adjusted general cognitive index at 4 years was significantly reduced in infants who were referred for neglect (-17.6; 95% confidence interval: -3.3, -31.9). Infants whose neglect was substantiated had a progressive decline in their cognitive function over time (cognitive z scores: -0.97, -1.37, and 2.05 standard deviations at 1, 2, and 4 years, respectively), compared with non neglected infants (z scores: -0.04 to -0.36). They had a significantly smaller head circumference at 2 and 4 years but not at birth (adjusted z score at 4 years: -0.812; 95% confidence interval: -0.167, -1.458). Perinatal risk factors and physical disability were not related to maltreatment referral; only parental factors were independent predictors. CONCLUSIONS: Childhood neglect is associated significantly with delayed cognitive development and head growth. Addressing risk factors antenatally and in early childhood may improve outcomes. PMID- 11433068 TI - Attitudes of parents and health care professionals toward active treatment of extremely premature infants. AB - OBJECTIVES: To compare the attitudes of neonatologists, neonatal nurses, the parents of extremely low birth weight (ELBW) children, and the parents of normal birth weight children toward saving infants of borderline viability and who should be involved in the decision-making process and to compare physicians' and nurses' estimates of the proportion of infants who are born at various gestational ages with regard to survival, morbidity, and treatment. METHODS: A questionnaire was given to 169 parents of ELBW children and 123 parents of term children, who were part of a longitudinal study of the outcome of ELBW infants. A similar questionnaire was completed by 98 Canadian neonatologists and 99 neonatal nurses. RESULTS: Physicians tended to be more optimistic than nurses regarding the probability of survival and freedom from serious disabilities and would recommend to parents life-saving interventions for their child at earlier gestational ages. A significant majority of parents believed that attempts should be made to save all infants, irrespective of condition or weight at birth, compared with only 6% of health professionals who endorsed this. In contrast to parents, health professionals believed that economic costs to society should be a factor in deciding whether to save an ELBW infant. However, health professionals did not believe that the economic status of the parents should be a factor, although the stress of raising an infant with disabilities should be. Most respondents believed that the parents and physicians should make the final decision but that other bodies, such as ethics committees or the courts, should not. CONCLUSION: Health care professionals must recognize that their attitudes toward saving ELBW infants differ from those of parents. Parents, whether of term or extremely premature children, are more in favor of intervening to save the infant irrespective of its weight or condition at birth than are professionals. It therefore is imperative that there be joint decision making, combining the knowledge of the physician with the wishes of the parents. PMID- 11433069 TI - Lead poisoning among refugee children resettled in Massachusetts, 1995 to 1999. AB - OBJECTIVE: Lead poisoning has been reported among immigrant and refugee populations in the United States; however, prevalences of elevated blood lead (BPb) and risk factors have not been described fully among newly arrived refugee children. This study was conducted to address this lack of data. METHODS: We performed analysis of BPb levels among a cohort of refugee children, aged <7 years, who arrived in Massachusetts between July 1, 1995, and December 31, 1999. Gender, age, birthplace, time of year of lead testing, intestinal parasitoses, anemia, and growth retardation were examined as predictors of elevated BPb. RESULTS: BPb levels >/=10 microgram/dL were found in 11.3% of 693 children shortly after their arrival in the United States. Children from developing countries had the highest prevalences, including 27% of Somalis and Vietnamese. Country of birth was the strongest predictor of elevated BPb. No association between elevated lead and age was found. Among 213 children with BPb tests >/=6 months after the initial test, 7% had newly elevated levels of >/=10 microgram/dL. CONCLUSIONS: The prevalence of elevated BPb levels in recently arrived refugee children is more than twice that of US-born children. Children who are at particular risk are those from developing countries where environmental exposures are more ubiquitous. In addition, a significant percentage of refugees acquired elevated levels after arrival, thus suggesting the importance of follow-up testing of refugee children. Refugee status should be considered a risk factor for lead poisoning. PMID- 11433070 TI - Sedation of children for electroencephalograms. AB - OBJECTIVE: Sedation sometimes is necessary to perform an electroencephalogram (EEG) on a child. A dramatic decline in the need to use conscious sedation in our EEG laboratory prompted this review of our sedation experience. The purpose of this review was to determine the incidence of adverse sedation effects and to determine why the need for sedation had declined. METHODS: All 513 attempts to administer sedation to children who were undergoing EEG studies during a 4-year period were reviewed retrospectively. Parameters studied included type and amount of the sedative agents, need for repeated dosing, successful completion of the EEG, and complications attributed to the sedative. RESULTS: Sedation was attempted in 513 (18%) of 2855 EEGs performed during the 4-year period. Ninety one percent of the EEGs performed with sedation were completed successfully. Chloral hydrate was the most frequently administered sedative. Complications (transient oxygen desaturation) occurred in 3 children, all of whom had recognized risk factors for airway compromise. The proportion of children who required sedation decreased from 32% to just 2% during that time period. CONCLUSION: Sedation of children who are undergoing EEG examinations is effective and safe. Complications are infrequent. The need for sedation can be decreased greatly by adequate preparation and by creating a less-threatening, child friendly environment in which to perform the study. PMID- 11433071 TI - Problems with prediction of neonatal hyperbilirubinemia. PMID- 11433072 TI - The risks of sedation for electroencephalograms: data at last. PMID- 11433073 TI - For a child, every moment is a teachable moment. PMID- 11433074 TI - Congenital autoimmune neutropenia in two premature neonates. AB - Autoimmune neutropenia (AIN) has been reported in infants and children, but not in neonates. AIN is caused by antibodies produced by the patient against their own neutrophils; therefore, it differs from the more common alloimmune neonatal neutropenia and the neonatal neutropenia because of a maternal autoimmune disease in which antineutrophil antibodies of maternal origin cross the placenta. We observed 2 cases of congenital AIN in premature neonates. These are the youngest reported cases, and indicate that AIN can have a prenatal onset. Examination of the bone marrow biopsies revealed an increase in B lymphocytes and myeloperoxidase-positive cells with a maturation arrest at the myelocyte stage. Recombinant human granulocyte colony-stimulating factor effectively treated the neutropenia, as it does in infantile AIN. Ten months after the diagnosis, 1 of the patients still requires recombinant human granulocyte colony-stimulating administration. PMID- 11433075 TI - Alcohol use and abuse: a pediatric concern. AB - Alcohol use and abuse by children and adolescents continue to be a major problem. Pediatricians should interview their patients regularly about alcohol use within the family, by friends, and by themselves. A comprehensive substance abuse curriculum should be integrated into every pediatrician's training. Advertising of alcohol in the media, on the Internet, and during sporting events is a powerful force that must be addressed. Availability of alcohol to minors must be controlled, and interventions for the child and adolescent drinker and punitive action for the purveyor are encouraged. PMID- 11433076 TI - Fireworks-related injuries to children. AB - An estimated 8500 individuals, approximately 45% of them children younger than 15 years, were treated in US hospital emergency departments during 1999 for fireworks-related injuries. The hands (40%), eyes (20%), and head and face (20%) are the body areas most often involved. Approximately one third of eye injuries from fireworks result in permanent blindness. During 1999, 16 people died as a result of injuries associated with fireworks. Every type of legally available consumer (so-called "safe and sane") firework has been associated with serious injury or death. In 1997, 20 100 fires were caused by fireworks, resulting in $22.7 million in direct property damage. Fireworks typically cause more fires in the United States on the Fourth of July than all other causes of fire combined on that day. Pediatricians should educate parents, children, community leaders, and others about the dangers of fireworks. Fireworks for individual private use should be banned. Children and their families should be encouraged to enjoy fireworks at public fireworks displays conducted by professionals rather than purchase fireworks for home or private use. PMID- 11433077 TI - Developmental surveillance and screening of infants and young children. AB - Early identification of children with developmental delays is important in the primary care setting. The pediatrician is the best-informed professional with whom many families have contact during the first 5 years of a child's life. Parents look to the pediatrician to be the expert not only on childhood illnesses but also on development. Early intervention services for children from birth to 3 years of age and early childhood education services for children 3 to 5 years of age are widely available for children with developmental delays or disabilities in the United States. Developmental screening instruments have improved over the years, and instruments that are accurate and easy to use in an office setting are now available to the pediatrician. This statement provides recommendations for screening infants and young children and intervening with families to identify developmental delays and disabilities. PMID- 11433078 TI - Technical report: mercury in the environment: implications for pediatricians. AB - Mercury is a ubiquitous environmental toxin that causes a wide range of adverse health effects in humans. Three forms of mercury (elemental, inorganic, and organic) exist, and each has its own profile of toxicity. Exposure to mercury typically occurs by inhalation or ingestion. Readily absorbed after its inhalation, mercury can be an indoor air pollutant, for example, after spills of elemental mercury in the home; however, industry emissions with resulting ambient air pollution remain the most important source of inhaled mercury. Because fresh water and ocean fish may contain large amounts of mercury, children and pregnant women can have significant exposure if they consume excessive amounts of fish. The developing fetus and young children are thought to be disproportionately affected by mercury exposure, because many aspects of development, particularly brain maturation, can be disturbed by the presence of mercury. Minimizing mercury exposure is, therefore, essential to optimal child health. This review provides pediatricians with current information on mercury, including environmental sources, toxicity, and treatment and prevention of mercury exposure. PMID- 11433079 TI - Shaken baby syndrome: rotational cranial injuries-technical report. AB - Shaken baby syndrome is a serious and clearly definable form of child abuse. It results from extreme rotational cranial acceleration induced by violent shaking or shaking/impact, which would be easily recognizable by others as dangerous. More resources should be devoted to prevention of this and other forms of child abuse. PMID- 11433080 TI - Adult outcomes of pediatric recurrent abdominal pain: do they just grow out of it? AB - OBJECTIVE: To determine whether medically unexplained recurrent abdominal pain (RAP) in childhood predicts abdominal pain, irritable bowel syndrome (IBS), other somatic complaints, and psychiatric symptoms and disorders in young adulthood. METHODS: A sample of 28 young adults evaluated for RAP between the ages of 6 and 17 years were compared with 28 individually matched former childhood participants in a study of tonsillectomy and adenoidectomy. RAP caseness was established by structured retrospective chart review requiring agreement by 2 independent reviewers. Standardized assessments of abdominal pain, IBS, other somatic symptoms, psychopathology, perceived health, and history of maltreatment were performed an average of 11.1 years after the index visit. RESULTS: Former RAP patients were significantly more likely than controls to endorse anxiety symptoms and disorders, hypochondriacal beliefs, greater perceived susceptibility to physical impairment, poorer social functioning, current treatment with psychoactive medication, and generalized anxiety in first degree relatives. There were trends suggesting associations between childhood RAP and lifetime psychiatric disorder, depression, migraine, and family history of depression, but group differences on abdominal pain, IBS, other somatic symptoms, and history of maltreatment were not statistically significant. CONCLUSIONS: There is a strong and relatively specific association between childhood RAP and anxiety in young adulthood. Affected children may be at special risk to perceive physical symptoms as threatening, and should be evaluated for psychiatric disorder on initial presentation. PMID- 11433081 TI - Scooter injuries: a new pediatric morbidity. AB - OBJECTIVE: To describe types of injuries, mechanisms of injury, and treatment of injuries caused by scooter use in children, and to discuss issues of injury prevention in children who use scooters. STUDY DESIGN: Data were collected from 14 children seen by a general pediatrician and an orthopedic surgeon over a 3 month period in the summer of 2000. Detailed histories were obtained from patients and their families, and medical records were reviewed. RESULTS: Eleven of the 14 patients suffered fractures. The injuries in the other 3 patients were a large abrasion, a laceration, and a septic knee. Half (7) of the children were injured within the first day of riding their scooter, and 13 of the 14 injuries occurred within the first month of scooter use. Only 5 patients used protective gear at the time of their injuries, and those patients were injured in unprotected parts of their bodies. CONCLUSIONS: The popularity of scooters presents a new cause of pediatric injuries and a significant health hazard to children. In our study, most injuries occurred shortly after children began scooter use, and younger children suffered the most severe injuries. Additional studies are needed to determine how scooter-related injuries can be prevented or minimized. scooters, injuries. PMID- 11433082 TI - Rational testing of the HIV-exposed infant. AB - OBJECTIVES: The objectives of this study were 1) to evaluate testing regimens of human immunodeficiency virus (HIV)-exposed infants and 2) to determine optimal methods of follow-up by enzyme-linked immunosorbent assay (ELISA) testing. METHODS: We reviewed the results from 742 HIV-exposed infants in the state of North Carolina; 2474 samples were tested for HIV by DNA polymerase chain reaction (PCR) at the University of North Carolina Retrovirology Core Laboratory. We then reviewed the utility and costs of ELISA testing of all HIV-exposed infants who were seen at the Duke University Pediatric Infectious Disease Clinic between January 1, 1993, and May 5, 1998. We used likelihood ratios to model probability of HIV infection given 3 negative DNA (PCR) tests and to provide recommendations on the use of ELISA follow-up. RESULTS: The overall sensitivity of the DNA PCR was 87.1%, and its specificity was 99.9%. We evaluated 224 HIV-exposed infants who were seen at Duke University and who had at least 3 negative diagnostic tests using either DNA PCR tests or HIV blood cultures. All 178 infants who subsequently underwent ELISA testing ultimately demonstrated seroreversion. The Duke University Pediatric Infectious Disease Clinic transferred the care of 65 patients to primary care physicians before ELISA testing and retained the care of the remaining 159 patients. Children who remained in Duke's care were more likely to have documentation of seroreversion (158 of 159 vs 20 of 65). We reviewed costs of travel, physician appointment, and HIV antibody testing in a tertiary care setting. Given 3 negative PCR tests, the expected cost per case of HIV detected by a positive ELISA assay is $23.8 million. CONCLUSIONS: Documentation of seroreversion in this cohort was nearly complete in the multidisciplinary subspecialty clinic but not when such responsibility was left to the primary care physician. Given the low probability of disease in patients who have had 3 negative PCR tests, documentation of a negative ELISA may not be an appropriate use of medical resources. PMID- 11433083 TI - Barriers to bicycle helmet use. AB - OBJECTIVE: To determine why people do or do not wear helmets while bicycling. METHODS: A survey was conducted from August through October 1999. Two survey areas were chosen for this study: local public schools and paved bicycle trails. For the school arm of the study, 3 public elementary, middle, and high schools were selected from 3 different regions of Rochester, Minnesota, for participation in the study. For the bicycle arm of the study, 3 paved trails located in southeastern Minnesota were selected. A total of 2970 surveys were distributed to the public school system, and 463 surveys were collected from bicyclists on the paved bicycle trails. The survey population was split into 3 age categories for analysis: child (7-10), adolescent (11-19), and adult (older than 19). RESULTS: Of the 2970 surveys distributed to Rochester public schools, 2039 (69%) were returned for analysis. Seventy-eight of the surveys that were completed in the public school system were discarded for the following reasons: age <10 years (35), insufficient completion (24), and selection of every reason for not wearing a bicycle helmet (19). A total of 463 surveys were completed on the 3 paved bicycle trails. One survey from the paved bicycle trail arm of the study was discarded because of insufficient completion. The total number of surveys used for statistical analysis was 2424. The distribution of male (52.7%) and female (47.3%) participants was similar. No significant difference in bicycle helmet use was found between genders. The age groups with the highest rate of bicycle helmet use were 50 to 59 years (62%) and older than 59 years (70%). The age groups with the lowest rate of bicycle helmet use were 11 to 19 years (31%) and 30 to 39 years (30%). The most common reasons given for not wearing a bicycle helmet were "uncomfortable," "annoying," "it's hot," "don't need it," and "don't own one." Bicycle helmet use was significantly influenced by peer helmet use in all 3 age groups. Children also were more likely to wear a bicycle helmet when their parents wore bicycle helmets. A majority of respondents in all 3 age groups indicated that bicycle helmets provided either "moderate" or "great" protection from head injury, although significantly more adults (65.9%) than adolescents (43.9%) believed that the protection afforded by bicycle helmets was "great." Despite this belief, a majority of adolescents and adults indicated that there was only a "slight risk" of head injury when bicycling without a helmet. Participants in all 3 categories were more likely to wear a bicycle helmet when they indicated either that there was a "great risk" of head injury when bicycling without a helmet or that helmets provided "great protection" from head injury. Adolescents and adults who believed that bicycling without a helmet put one at "great risk" for head injury also were more likely to indicate that helmets provided "great protection" from head injury. CONCLUSIONS: The prevalence of bicycle helmet use remains low despite research indicating the high level of head injury risk when bicycling without a helmet and the significant protection afforded by bicycle helmets. With the information provided by this survey, a well designed intervention to increase the use of bicycle helmets can be implemented. Suggestions for a campaign to promote an increase in bicycle helmet use include focusing efforts on males and females between 11 and 19 years and 30 and 39 years of age; educating the public on new bicycle helmet designs that address comfort, ventilation, and fashion; educating adolescents on the significant protection from head injury afforded by bicycle helmets; and educating the public on the risk and severity of head injury associated with bicycling without a helmet. The influence of parents and peers on bicycle helmet use may be targeted through education and statements such as, "If you wear a bicycle helmet, you are not only protecting yourself, you are also helping to protect your friends and/or children." bicycle, helmet, injury, accident, prevention. PMID- 11433084 TI - Hypokalemic salt-losing tubulopathy with chronic renal failure and sensorineural deafness. AB - OBJECTIVE: To characterize a rare inherited hypokalemic salt-losing tubulopathy with linkage to chromosome 1p31. METHODS: We conducted a retrospective analysis of the clinical data for 7 patients in whom cosegregation of the disease with chromosome 1p31 had been demonstrated. In addition, in 1 kindred, prenatal diagnosis in the second child was established, allowing a prospective clinical evaluation. RESULTS: Clinical presentation of the patients was homogeneous and included premature birth attributable to polyhydramnios, severe renal salt loss, normotensive hyperreninemia, hypokalemic alkalosis, and excessive hyperprostaglandin E-uria, which suggested the diagnosis of hyperprostaglandin E syndrome/antenatal Bartter syndrome. However, the response to indomethacin was only poor, accounting for a more severe variant of the disease. The patients invariably developed chronic renal failure. The majority had extreme growth retardation, and motor development was markedly delayed. In addition, all patients turned out to be deaf. CONCLUSION: The hypokalemic salt-losing tubulopathy with chronic renal failure and sensorineural deafness represents not only genetically but also clinically a disease entity distinct from hyperprostaglandin E syndrome/antenatal Bartter syndrome. A pleiotropic effect of a single gene defect is most likely causative for syndromic hearing loss. PMID- 11433085 TI - Thiazide diuretics arrest the progression of nephrocalcinosis in children with X linked hypophosphatemia. AB - OBJECTIVE: X-linked hypophosphatemia (XLH) is characterized clinically by rickets, hypophosphatemia, and hyperphosphaturia. Conventional treatment of XLH with oral phosphate and vitamin D is associated with increased urinary calcium excretion and nephrocalcinosis. Thiazide diuretics decrease urinary calcium excretion. The objective of this study was to determine the effect of thiazide diuretics on the clinical and radiologic course of nephrocalcinosis in children with XLH. METHODS: The effect of hydrochlorothiazide (HCTZ) on clinical and radiologic progression of nephrocalcinosis was evaluated in 11 children with XLH. All patients had been treated previously with vitamin D and oral phosphate and had radiologic evidence of nephrocalcinosis. The average age of the patients at the start of HCTZ was 6.6 +/- 1.0 years. The effect of oral HCTZ at 0.8 +/- 0.1 mg/kg body weight per day given for 3.3 +/- 0.6 years on the progression of nephrocalcinosis and urinary calcium excretion was evaluated. RESULTS: There was no change in serum phosphorous, calcium, potassium, and chloride after HCTZ therapy. HCTZ therapy increased serum bicarbonate and decreased urinary calcium excretion. The grade of nephrocalcinosis increased from 0.4 +/- 0.2 to 1.5 +/- 0.3 in the 2.3 +/- 0.3 years before initiation of HCTZ therapy, whereas the degree of nephrocalcinosis was stable after 3.3 +/- 0.6 years of HCTZ therapy (1.5 +/- 0.3 vs 3.0 +/- 0.3). CONCLUSION: HCTZ decreased urinary calcium excretion but did not result in the resolution of nephrocalcinosis. However, when compared with the control period, HCTZ prevented the progression of nephrocalcinosis in children with XLH. PMID- 11433086 TI - Fetal surgery for posterior urethral valves: long-term postnatal outcomes. AB - OBJECTIVE: Fetal intervention for obstructive uropathy was first performed at the University of California, San Francisco in 1981. Indications for treatment were bilateral hydronephrosis with oligohydramnios. Preintervention criteria included fetal urinary electrolytes with beta-microglobulin levels, karyotyping, and detailed sonography specifically looking for renal cortical cysts. We reviewed the outcomes of children who underwent fetal intervention with specific long-term follow-up in patients who were found postnatally to have posterior urethral valves. METHODS: A retrospective review of the University of California, San Francisco fetal surgery database was performed for patients with a prenatal diagnosis of obstructive uropathy. Medical records from 1981 to 1999 were reviewed. Long-term follow-up was documented if the cause of the urinary tract obstruction was posterior urethral valves. We collected data points, focusing on time and type of intervention, fetal urinary electrolytes, appearance of fetal kidneys, present renal function, length of follow-up, and present status of the urinary tract. RESULTS: Forty patients were evaluated for fetal intervention; 36 fetuses underwent surgery during this time period. Postnatal confirmation of posterior urethral valves was demonstrated in 14 patients. All patients had favorable fetal urinary electrolytes. Mean gestational age at intervention was 22.5 weeks. The procedures performed included creation of cutaneous ureterostomies in 1, fetal bladder marsupialization in 2, in utero ablation of valves in 2, and placement of vesicoamniotic catheter in 9. Six deaths occurred before term delivery with premature labor and the newborns succumbing to respiratory failure. One pregnancy was terminated electively because of shunt failure and declining appearance of fetal lungs and kidney. The remaining 8 living patients had a mean follow-up of 11.6 years. Chronic renal disease with abnormal serum creatinine was present in 5 patients. Two patients have undergone renal transplantation, and 1 is awaiting organ donation. Five of the 8 living patients have had urinary diversion with vesicostomy, cutaneous ureterostomy, or augmentation cystoplasty with later reconstruction. CONCLUSIONS: Fetal intervention for posterior urethral valves carries a considerable risk to the fetus with fetal mortality rate of 43%. The long-term outcomes indicate that intervention may not change the prognosis of renal function or be a predictor for possible urinary diversion. Despite all of these patients' having favorable urinary electrolytes, this did not seem to have any implication postnatally. When counseling families about fetal intervention, efforts should be focused on that intervention may assist in delivering the fetus to term and that the sequelae of posterior urethral valves may not be preventable. Fetal surgery for obstructive uropathy should be performed only for the carefully selected patient who has severe oligohydramnios and "normal"-appearing kidneys. PMID- 11433087 TI - Pediatric residency training on tobacco. AB - OBJECTIVE: Pediatricians have a unique role to play in the antismoking arena. However, few pediatric residency training programs prepare residents to meet the tobacco challenge. This study evaluates the effects of a comprehensive pediatric residency training program on tobacco on resident tobacco intervention behaviors, as well as on changes in the behavior of patients and their parents/guardians. METHODS: Pediatric residents were exposed to a comprehensive training program on tobacco. Baseline and follow-up surveys of residents, parents/guardians, and patients were used to assess the effects of the training program. A quasi experimental design permitted unambiguous evaluation of the program's effects on resident intervention behaviors. RESULTS: The comprehensive training program on tobacco led to marked and significant changes in resident intervention on tobacco. Many of these changes were supported by parents' and patients' reports. In turn, resident intervention on tobacco led to a significant increase in the likelihood that parents would maintain a "smoke-free household." Significant changes in the prevalence of parental smoking were not found, although the trend during 3 years of follow-up was in the desired direction. A high proportion of residents reported that they intervened on tobacco in patients at baseline and follow-up, but too few patients were sampled to permit analysis of the impact on changes in smoking. CONCLUSIONS: Comprehensive training on tobacco had a positive and powerful effect on the tobacco intervention behavior of pediatric residents. In turn, intervention on tobacco by pediatric residents may have a significant impact on patients and their parents. These findings underscore the efficacy of pediatric residency training on tobacco, and it is hoped that they will serve as an impetus for other pediatric residency programs to introduce training on tobacco. PMID- 11433088 TI - The effect of anorectal manometry on the outcome of treatment in severe childhood constipation: a randomized, controlled trial. AB - OBJECTIVE: Approximately 50% of constipated children contract rather than relax the external sphincter complex during a defecation attempt. Although biofeedback training (BF) is able to change this defecation behavior, there is no additional effect of BF to conventional treatment (CT) on clinical outcome compared with CT alone. It has been postulated that the absence of a significant difference between these 2 treatment options might be because of a therapeutic, "demystifying" effect of performing anorectal manometry in conventionally treated children, necessary to obtain basal manometric data. The objective of this prospective, controlled, randomized study was to evaluate the effect of CT with 2 anorectal manometry sessions compared with CT alone (dietary advice, diary, toilet training, oral laxatives, and enemas) on clinical outcome. METHODS: A total of 212 constipated children (143 boys) who were visiting a referral pediatric gastroenterologic practice were randomized prospectively to CT alone (115 patients) or to CT combined with 2 manometry sessions (CTM; 97 patients). Patients were included in the study when they fulfilled at least 2 of the 4 following criteria: stool frequency fewer than 3 per week, 2 or more soiling and/or encopresis episodes per week, periodic passage of very large amounts of stool every 7 to 30 days, or a palpable rectal or abdominal fecal mass. CT comprises dietary advice, a daily diary, toilet training, and oral laxative treatment preceded by rectal disimpaction with enemas on 3 consecutive days. During both manometries, the child and the parent could watch the tracing on the computer screen. No explanation was given to either the child or the parents during the procedure. When the procedure was finished, the tracings were clarified. Successful treatment was defined as a defecation frequency of 3 or more per week and fewer than 1 soiling/encopresis episode per 2 weeks and no use of laxatives. RESULTS: Only 4 and 2 children from the CT and CTM groups showed no soiling and/or encopresis, whereas 76% and 65%, respectively, reported the periodic passage of large stools. In 26% and 30% of the patients, a rectal scybalum was found on physical examination. The success rates at 6, 26, 52, and 104 weeks' follow-up were 4%, 24%, 32%, and 43% and 7%, 22%, 30%, and 35% in the CT and CTM group, respectively. No significant difference in success percentage was observed between the 2 groups at any time of follow-up with relative risks (CT/CTM) and 95% confidence intervals, respectively, of 0.55 (0.16-1.89), 1.13 (0.67-1.89), 1.07 (0.69-1.65), and 1.23 (0.81-1.85). A significant increase in defecation frequency was observed between the first (intake) and second visits, which was sustained at all subsequent visits and stages of follow-up in both groups (not significant). Also in relation to the first visit, a significant decrease in encopresis episodes was shown and a further slow but significant decrease at 52 weeks of follow-up in both groups. The manometric data obtained from the CTM group showed a low percentage of children with normal defecation dynamics, namely 28%, which (significantly) increased to 38% at the last manometry. CONCLUSIONS: Anorectal manometry combined with CT compared with CT alone did not result in higher success rates in chronically constipated children. Therefore, anorectal manometry has no additional demystifying or educational effect on clinical outcome in chronically constipated children. This observation together with the observation in the current and previous studies that no correlation was found between (achievement of) normal defecation dynamics and success and that no relation was observed between volume of urge or critical volume and success leaves no diagnostic or therapeutic role for anorectal manometry in chronic constipated children, except its use as a diagnostic test to exclude Hirschsprung's disease. A simple CT is successful in 30% of severely constipated children who are referred to a tertiary hospital, underscoring the importance of long-lasting and adequate laxative treatment. PMID- 11433089 TI - Children who return home from foster care: a 6-year prospective study of behavioral health outcomes in adolescence. AB - OBJECTIVE: Returning children to their biological families after placement in foster care (ie, reunification) has been prioritized with legislation. Comprehensive studies of child behavioral health functioning after reunification, however, have not been conducted. This study examined outcomes for youth who were reunified after placement in foster care as compared with youth who did not reunify. DESIGN: Prospective cohort. SETTING: Children who entered foster care in San Diego, California, and who remained in foster care for at least 5 months. Participants. A cohort of 149 ethnically diverse youth, 7 to 12 years old, who entered foster care between May 1990, and October 1991. Seventy-five percent of those interviewed at Time 1 were interviewed at Time 2 (6 years later). OUTCOME MEASURES: 1) Risk behaviors: delinquent, sexual, self-destructive, substance use, and total risk behaviors; 2) Life-course outcomes: pregnancy, tickets/arrests, suspensions, dropping out of school, and grades; 3) Current symptomatology: externalizing, internalizing, total behavior problems, and total competence. RESULTS: Compared with youth who were not reunified, reunified youth showed more self-destructive behavior (0.15 vs -0.11), substance use (0.16 vs -0.11), and total risk behavior problem standardized scores (0.12 vs -0.09). Reunified youth were more likely to have received a ticket or have been arrested (49.2% vs 30.2%), to have dropped out of school (20.6% vs 9.4%), and to have received lower grades (6.5 vs 7.4). Reunified youth reported more current problems in internalizing behaviors (56.6 vs 53.0), and total behavior problems (59.5 vs 55.7), and lower total competence (41.1 vs 45.0). There were no statistically significant differences between the groups on delinquency, sexual behaviors, pregnancy, suspensions, or externalizing behaviors. Reunification status was a significant predictor of negative outcomes in 8 of the 9 regression equations after controlling for Time 1 behavior problems, age, and gender. CONCLUSIONS: These findings suggest that youth who reunify with their biological families after placement in foster care have more negative outcomes than youth who do not reunify. The implications of these findings for policy and practice are discussed. PMID- 11433090 TI - Navajo perceptions of asthma and asthma medications: clinical implications. AB - BACKGROUND: Despite the large number of asthma patients, relatively little is known about the beliefs of asthmatic children and their parent(s), or the extent to which these beliefs influence management of the disease. OBJECTIVE: The purpose of this study was to examine how medication-taking and the use of health care services were influenced by perceptions of the disease and beliefs about medications among Navajo families. METHODS: Ethnographic interviews were used to investigate the disease and medication beliefs of 22 Navajo families with 29 asthmatic children. RESULTS: Most respondents perceived asthma as the transient experience of symptoms in someone who was chronically vulnerable to breathing problems. The majority (97%) of these asthmatics reported using bronchodilators, although only 34% reported current use of antiinflammatory medications. Although controller medications were distinguished from rescue medications, they were thought to have the same effect on the lungs. Many families were concerned about becoming dependent on the medicines and attempted to "wean" the asthmatic from the drugs. A large proportion (80%) of children had assumed responsibility for their own medication taking. Parents often referred to previous attacks requiring treatment in the emergency department when deciding whether to initiate medication or seek medical attention, often resulting in a delay of treatment. Nebulized treatments delivered in the emergency department were perceived to be the strongest therapy available. As a result, visits to the emergency department were common (79%), as were hospital admissions (57%). CONCLUSION: This study provides evidence that, among pediatric Navajo asthma patients, perceptions of asthma and beliefs about the activity of asthma medications influence when and how often asthma medicines are taken, as well as the use of health care services. In addition, excessive reliance on emergency treatments and the high rates of hospital admissions suggest that asthma is undertreated in this group of Navajo patients. PMID- 11433091 TI - Neutrophil CD11b expression and circulating interleukin-8 as diagnostic markers for early-onset neonatal sepsis. AB - OBJECTIVE: To assess neutrophil CD11b and circulating interleukin 8 (IL-8) as markers of early-onset infection in neonates. METHODS: The study comprised 39 neonates, with a gestational age of 29 to 41 weeks, suspected of infection within 48 hours of life. Neutrophil surface expression of CD11b was quantified with flow cytometry and plasma IL-8 with an enzyme-linked immunosorbent assay. Both data were available from 35 of 39 neonates. Serum C-reactive protein was determined at initial evaluation and, later, on the basis of the clinical picture. Neonates were allocated retrospectively into 2 groups. In the sepsis group (N = 22), 4 had culture-proven sepsis, and 14 had an antenatal risk factor for infection. In the possible-infection group (N = 13), each neonate had a noninfective disorder, but co-occurring infection remained a possibility. Twelve healthy term infants served as controls. RESULTS: CD11b expression and IL-8 levels both increased in order of sepsis > possible infection > healthy. Sensitivity and specificity by the CD11b test for sepsis were equal, at 1.00, and those by the IL-8 test 0.91 and 1.00, respectively; 6 (17.1%) of the 35 neonates had CD11b and IL-8 below cutoff levels. CONCLUSIONS: Measuring neutrophil CD11b expression and circulating IL-8 provides a means to identify early-onset neonatal sepsis. The findings may be helpful in planning strategies to safely reduce the use of antimicrobials in neonates. PMID- 11433092 TI - Rapid assessment of sexual behavior, drug use, human immunodeficiency virus, and sexually transmitted diseases in northern thai youth using audio-computer assisted self-interviewing and noninvasive specimen collection. AB - BACKGROUND: Drug use, unwanted pregnancy, human immunodeficiency virus (HIV) infection, and sexually transmitted diseases are serious health problems among Thai youth. The gravity of these problems demands high-quality data to direct public health policy and prevention programs. Previous studies of stigmatized behaviors have been hampered by participation bias and underreporting. To evaluate sexual behavior, disease, and drug use, we used audio-computer-assisted self-interviewing (ACASI) and noninvasive specimen collection methods. We also evaluated effectiveness of these methods in minimizing participation bias and underreporting. METHODS: In late 1999, students aged 15 to 21 years attending 3 vocational schools were invited to participate in a cross-sectional survey. Consenting students completed a classroom-based ACASI interview using a confidential code number system. Oral fluid specimens were tested for HIV antibodies, and urine was tested for chlamydial and gonococcal nucleic acids, methamphetamines, and opiates. RESULTS: Of 1736 invited students, 1725 (99%) agreed to participate. Of these, 48% of the male students and 43% of the female students reported ever having had sexual intercourse. Overall, the mean number of lifetime sexual partners was 4.6 among male participants (median: 2) and 2.8 among female participants (median: 1). Consistent use of condoms with steady partners was reported by 16% of male participants and 11% of female participants who had such partners. Of all male participants, 7% had ever paid for sex, 3% had ever sold sex, and 7% had ever been coerced to have sex. Of all female participants, 3% had ever sold sex and 21% had ever been coerced to have sex. Among women with a history of sexual intercourse, 27% reported at least 1 pregnancy. Of these pregnancies, 83% were terminated. Among those with sexual intercourse experience, the prevalence of HIV infection was 0.5%; of infection with Neisseria gonorrhoeae, 0.4%; and of infection with Chlamydia trachomatis, 5%. Twenty-nine percent of students reported ever having used methamphetamines. Ten percent had a methamphetamine-positive urine test. In the ACASI interview, 16% of these denied ever having used methamphetamines. The prevalence of opiate positive urine tests was low (0.2%). CONCLUSION: This study shows that adolescents and young adults in Chiang Rai are at high risk for having unprotected intercourse, being coerced to have sex, unwanted pregnancy, sexually transmitted diseases, and drug use. The high enrollment rate demonstrates the feasibility and acceptability of using ACASI and noninvasive specimen collection methods in a developing country. ACASI use may lead to increased, but not to complete, self-reporting of sensitive behaviors. PMID- 11433093 TI - Obsessive-compulsive scale of the child behavior checklist: specificity, sensitivity, and predictive power. AB - OBJECTIVE: To create an obsessive-compulsive disorder subscale (OCS) of the Child Behavior Checklist (CBCL) and to determine its internal consistency, sensitivity, specificity, and positive and negative predictive power to identify obsessive compulsive disorder (OCD) in children and adolescents. METHODS: Three samples of equal size (n = 73) of children and adolescents, matched for age, gender, and race, were selected for these analyses: 1) a clinically ascertained OCD group, 2) a psychiatrically treated group whose records revealed no evidence of OCD, and 3) a general population control group. An OCS was created by applying factor analysis to 11 CBCL items. Examinations of internal consistency, sensitivity, specificity, and positive and negative predictive value were undertaken. RESULTS: Of 11 items hypothesized to predict OCD, 8 items were retained after factor analyses (smallest factor loading: 0.49) and used to calculate OCS scores. The retained items displayed excellent internal consistency (Cronbach's alpha coefficient = 0.84). OCD participants had significantly higher OCS scores than either psychiatrically treated or general population control groups. With the use of the 2 cutoff scores closest to the true rate of OCD in the overall sample, sensitivity was 75.3% to 84.9%, specificity was 82.2% to 92.5%, positive predictive value was 70.5% to 83.3%, and negative predictive value was 88.2% to 91.6%. CONCLUSION: The performance of the proposed CBCL OCS compares favorably with that of the only previously studied screening instrument for OCD, the Leyton Obsessional Inventory-Child Version. Unlike the Leyton Obsessional Inventory Child Version, the CBCL is already in widespread use as a screen for most other forms of psychopathology. As the performance of the CBCL OCS will need to be replicated in other sample populations, data with various cutoff levels are provided to enable investigators and clinicians to tailor its use to specific study populations. PMID- 11433094 TI - Leptin levels in preterm human breast milk and infant formula. AB - OBJECTIVE: Leptin, a hormone present in breast milk, is involved in energy regulation and metabolism. The purpose of this investigation was to determine whether leptin is present in either preterm breast milk (PBM) or preterm formula (PF). The effects of delivery methods and pasteurization on leptin levels also were evaluated. METHODS: PBM samples were obtained from 29 mothers who delivered infants at between 23 and 34 weeks' gestation. Leptin levels were measured in PBM and PF with the use of a radioimmunoassay specific for human and bovine leptin, respectively. Milk samples were pasteurized by fast- and slow-heating methods. PBM and PF spiked with human leptin were delivered through catheters by bolus and continuous administration to determine the effects of delivery method on recoverable leptin levels. RESULTS: Median PBM leptin concentration was 5.28 ng/mL (intraquartile range: 24.79). Birth gestational age, birth weight, and gender of the infant did not significantly influence PBM leptin levels. Neither bolus nor continuous feeding practices affected leptin levels in PBM or spiked PF. However, pasteurization significantly reduced the amount of detectable leptin in PBM. CONCLUSIONS: PBM leptin levels were highly variable and similar to levels reported for term breast milk. There was no effect of postnatal age on PBM leptin concentrations. Sterilization decreased detectable leptin levels, whereas feeding practices had no adverse effect on the quantity of leptin delivered. Although no infant formula contained leptin, leptin could be added to formula and delivered through various feeding methods without loss. PMID- 11433095 TI - Staphylococcal scalded-skin syndrome in a very low birth weight premature infant. AB - Exfoliative skin diseases are rare in neonates. When caused by coagulase-positive Staphylococcus aureus, scalded-skin diseases such as staphylococcal scalded-skin syndrome (SSSS), bullous impetigo, and staphylococcal scarlet fever may develop. These diseases might cause significant complications and mortality. SSSS is caused by staphylococcal exfoliative toxins A or B, which split the granular layer of the skin, induce proteolysis, and might exhibit superantigen activities, such as epidermolysis and lymphocyte mitogenicity. We describe a 1378-g premature male infant who was born at 29 weeks' gestation and developed SSSS on day 3 of life, with no clinical signs of neonatal sepsis. After cultures from the lesion and bloodstream were obtained, intravenous cloxacillin therapy was started. Infection control measures were implemented instantly and included isolation of the infected infant, personnel handwashing with hexachlorophene, and placement of exposed neonates into a cohort. The initial lesion expanded and additional lesions appeared, but 12 hours after initiation of antibacterial therapy, the lesions ceased to proliferate. Cultures from scalded-skin lesions grew coagulase positive Staphylococcus aureus, whereas the bloodstream culture was sterile. The lesions resolved completely within 6 days, and the infant's subsequent course was uneventful. No similar skin lesions were noticed in other infants in the neonatal intensive care unit. We discuss recent advances in understanding the pathogenesis of neonatal SSSS, highlight the importance of early diagnosis and treatment, and stress the need for new adjunctive therapies for this disease. PMID- 11433096 TI - Recovery of anaerobic bacteria from four children with postthoracotomy sternal wound infection. AB - The cases of 4 children who developed postthoracotomy sternal wound infection caused by anaerobic bacteria are presented. The predominate anaerobes were Peptostreptococcus species and pigmented Prevotella species. Polymicrobial infection was present in all cases, and aerobic bacteria also were recovered in 2 instances. All patients responded to surgical debridement and antimicrobials effective against the isolated aerobic and anaerobic bacteria. These findings highlight the potential importance of anaerobic bacteria in postthoracotomy sternal wound infection. PMID- 11433097 TI - Invasive serotype a Haemophilus influenzae infections with a virulence genotype resembling Haemophilus influenzae type b: emerging pathogen in the vaccine era? AB - OBJECTIVE: Haemophilus influenzae type b causes severe disease in nonimmune infants and young children; other serotypes are uncommon pathogens and thought to have low virulence. Some have hypothesized that with the virtual elimination of H influenzae type b, other serotypes might acquire virulence traits and emerge as important pathogens of children. We describe the clinical, epidemiologic, and molecular biologic features of 5 cases of severe disease attributable to Haemophilus influenzae type a. METHODS: After observing 4 cases of invasive disease caused by H influenzae type a, we reviewed microbiology records at 3 reference laboratories that perform all serotyping in Utah and surveillance databases. Strains of H influenzae type a and control strains were examined by Southern blotting with the use of the cap probe pUO38 and by pulsed-field gel electrophoresis. The putative virulence mutation, the IS1016-bexA deletion, was detected by polymerase chain reaction amplification and sequencing. RESULTS: During a 10-month period, we observed 5 children with severe invasive disease caused by H influenzae type a. No isolates of H influenzae type a had been submitted to the reference laboratories between 1992 and 1998. The median age of patients was 12 months (range: 6-48 months). Four of 5 had meningitis and bacteremia; 1 had purpura fulminans. Three isolates, representing 1 of 2 pulsed field gel electrophoresis patterns, contained the IS1016-bexA deletion and were associated with particularly severe disease. CONCLUSIONS: We describe an unusual cluster of severe disease caused by H influenzae type a that resembles the clinical and epidemiologic features of H influenzae type b disease. Our data support the hypothesis that the IS1016-bexA deletion may identify more virulent strains of H influenzae. Haemophilus influenzae, epidemiology, virulence, serotyping, pathogenicity. PMID- 11433098 TI - Diagnosis of very long chain acyl-dehydrogenase deficiency from an infant's newborn screening card. AB - Very long chain fatty acid dehydrogenase (VLCAD) deficiency is a rare but treatable cause of cardiomyopathy, fatty liver, skeletal myopathy, pericardial effusions, ventricular arrhythmias, and sudden death. Unrecognized, VLCAD deficiency may be rapidly progressive and fatal, secondary to its cardiac involvement. Because early diagnosis improves outcome, we present a neonate with VLCAD deficiency in whom retrospective analysis of the newborn screening card revealed that a correct diagnosis could have been made by newborn screening using tandem mass spectrometry. Our patient demonstrated a classic neonatal course with transient hypoglycemia at birth, interpreted as culture-negative sepsis, followed by a quiescent period notable only for hypotonia and poor feeding. At 3 months, he presented with cardiorespiratory failure and pericardial effusions, requiring pericardiocentesis, tracheostomy, and prolonged mechanical ventilation. Plasma free-fatty acid and acylcarnitine profiles demonstrated small but significant elevations of C14:2, C14:1, C16, and C18:1 acylcarnitine species, findings consistent with a biochemical diagnosis of VLCAD deficiency. Enteral feeds were changed to Portagen formula with marked improvement in cardiac symptoms over several weeks. To confirm the biochemical diagnosis, molecular analysis was performed by analysis of genomic DNA on a blood sample of the patient. Sequencing analysis and delineation of VLCAD mutations were performed using polymerase chain reaction and genomic sequencing. The patient was heterozygous for 2 different disease-causing mutations at the VLCAD locus. The maternal mutation was a deletion of bp 842-3 in exon 8, causing a shift in the reading frame. The paternal mutation was G+1A in the consensus donor splice site after exon 1; this splice-site mutation would likely result in decreased mRNA. The likely consequence of these mutations is essentially a null phenotype. To determine whether this case could have been picked up by tandem mass spectrometry analysis at birth when the patient was asymptomatic, acylcarnitine analysis was performed on the patient's original newborn card (after obtaining parental consent, the original specimen was provided courtesy of Dr Kenneth Pass, Director, New York State Newborn Screening Program). The blood sample had been obtained at 1 week of age and stored at room temperature for 6 months and at 70 degrees C thereafter for 18 months. Electrospray tandem mass spectrometry used a LC-MS/MS API 2000 operated in ion evaporation mode with the TurboIonSpray ionization probe source. The acylcarnitine profile obtained from the patient's original newborn card was analyzed 2 years after it was obtained. In comparison with a normal control, there was a significant accumulation of long chain acylcarnitine species, with a prominent peak of tetradecenoylcarnitine (C14:1), the most characteristic metabolic marker of VLCAD deficiency. This profile would have likely been even more significant if it had been analyzed at the time of collection, yet 2 years later is sufficient to provide strong biochemical evidence of the underlying disorder. Discussion. VLCAD was first discovered in 1992, and clinical experience with VLCAD deficiency has been accumulating rapidly. Indeed, the patients originally diagnosed with long chain acyl-CoA deficiency suffer instead from VLCAD deficiency. The phenotype of VLCAD deficiency is heterogeneous, ranging from catastrophic metabolic and cardiac failure in infancy to mild hypoketotic, hypoglycemia, and exertional rhabdomyolysis in adults. This case demonstrates that VLCAD deficiency could have been detected from the patient's own neonatal heel-stick sample. Most likely, a presymptomatic diagnosis would have avoided at least part of a lengthy and intensive prediagnosis hospitalization that had an estimated cost of $400 000. Although VLCAD is relatively rare, timely and correct diagnosis leads to dramatic recovery, so that detection by newborn screening could prevent the onset of arrhythmias, heart failure, metabolic insufficiency, and death. Fatty acid oxidation defects, including VLCAD deficiency, may account for as many as 5% of sudden infant death patients. Recent instrumentation advances have made automated tandem mass spectrometry of routine neonatal heel stick samples technically feasible. Pilot studies have demonstrated an incidence of fatty acid oxidation defects, including short chain, medium chain, and very long chain acyl-CoA dehydrogenase deficiencies, of approximately 1/12 000. As a result, cost-benefit ratios for this approach should be systematically examined. PMID- 11433101 TI - Consumer & family information. Schizophrenia. PMID- 11433099 TI - Can the public sector survive managed care? PMID- 11433102 TI - Datapoints: Trends in psychiatric practice, 1988-1998: II. Caseload and treatment characteristics. PMID- 11433103 TI - Clinical computing: Use of a personal digital assistant in reducing medication error rates. PMID- 11433104 TI - Law and psychiatry: Liability for forensic evaluations: a word of caution. PMID- 11433105 TI - Rehab rounds: Social skills training to help mentally ill persons find and keep a job. PMID- 11433106 TI - Managed care: Toward open-source practices in managed behavioral health. PMID- 11433107 TI - Evidence-based practices for services to families of people with psychiatric disabilities. AB - Family psychoeducation is an evidence-based practice that has been shown to reduce relapse rates and facilitate recovery of persons who have mental illness. A core set of characteristics of effective family psychoeducation programs has been developed, including the provision of emotional support, education, resources during periods of crisis, and problem-solving skills. Unfortunately, the use of family psychoeducation in routine practice has been limited. Barriers at the level of the consumer and his or her family members, the clinician and the administrator, and the mental health authority reflect the existence of attitudinal, knowledge-based, practical, and systemic obstacles to implementation. Family psychoeducation dissemination efforts that have been successful to date have built consensus at all levels, including among consumers and their family members; have provided ample training, technical assistance, and supervision to clinical staff; and have maintained a long-term perspective. PMID- 11433108 TI - Diagnosis and treatment of mood disorders that co-occur with schizophrenia. AB - Affective syndromes, particularly mania and depression, often occur during the course of illness in patients who have a diagnosis of schizophrenia. Our current diagnostic system allows clear classification of the overlap of affective symptoms and psychotic symptoms. However, this diagnostic system will eventually require major revisions so that it will more accurately reflect the biologic mechanisms that underlie different symptoms. This article reviews the historical underpinnings of the diagnostic classification of affective and psychotic symptoms, including a discussion of current diagnostic practices and a review of the few studies on treatment of affective symptoms and syndromes in patients with schizophrenia. There is a great need for additional research on treatment of mood disorders that arise during the course of schizophrenia, especially with regard to newer mood stabilizers and antidepressants. PMID- 11433109 TI - Impact of panic attacks on rehabilitation and quality of life among persons with severe psychotic disorders. AB - OBJECTIVE: The study evaluated data from a sample of persons with severe psychotic disorders to determine whether those with and without comorbid panic attacks differed in rates of comorbidity of other psychiatric disorders, in quality of life, and in rehabilitation outcomes. METHODS: A total of 120 individuals with psychotic disorders were assessed with the Center for Epidemiologic Studies-Depression scale, the Structured Clinical Interview for DSM III-R, the General Health Questionnaire, the Global Assessment of Functioning scale, and several quality-of-life measures at baseline and four and a half months after they had participated in a social rehabilitation program. Multivariate analyses of variance and Pearson's chi square tests were used to compare baseline and follow-up scores between individuals who did and did not have panic attacks. RESULTS: Eighteen (15 percent) of the participants who had severe psychotic disorders also had panic attacks. Participants with this type of comorbidity had significantly higher rates of major depressive disorder, specific phobia, sedative abuse, polysubstance abuse, other substance abuse, and anorexia nervosa than participants who did not have panic attacks. Participants who had panic attacks also had poorer rehabilitative outcomes and poorer quality of life at baseline and at follow-up than participants who did not have panic attacks. CONCLUSIONS: These data are the first to show that comorbid panic attacks are associated with poorer rehabilitative outcomes and poorer quality of life among individuals with severe psychotic disorders than among those who have psychotic disorders without panic attacks. Panic attacks may be a valuable prognostic indicator among persons with psychotic disorders and may have implications for treatment and rehabilitation. PMID- 11433110 TI - Pushing the quality envelope: a new outcomes management system. AB - This article is based on the authors' experience in designing and implementing outcomes management systems for large managed care organizations. Topics addressed include design of instruments, use of cost-effective technology, development of computerized decision-support tools, and methods for case-mix adjustment. The case-mix-adjustment models are based on a data repository of several thousand treatment cases with multiple measurement points across the course of treatment. Data from controlled and field studies are described. These data suggest that the outcomes management methods outlined in this article can result in significantly improved clinical outcomes and a more rational allocation of behavioral health care resources. PMID- 11433111 TI - Predictors of dissemination of family psychoeducation in community mental health centers in Maine and Illinois. AB - OBJECTIVE: The aim of this study was to determine whether it is possible at the time of staff training to predict whether a mental health center will succeed in implementing family psychoeducation services. METHODS: Fifteen mental health agencies in Maine and 51 in Illinois in which clinicians and administrators were trained in multifamily psychoeducation were studied. Participants were surveyed immediately after their initial training sessions and nine months later. Participants' demographic characteristics, agency characteristics, and principal components derived from the survey data were analyzed to identify factors associated with implementation of family psychoeducation services. RESULTS: The regression model successfully predicted which sites would succeed. Success was more likely at sites where the treatment model was viewed more positively at the outset, where real and perceived resource limitations were addressed, where inducements to implementation were seen as less important, and where attention was paid to the difference between new and existing treatment methods. Nearly all the Maine sites (14, or 93 percent) implemented multifamily psychoeducation services, whereas only five of the Illinois sites (10 percent) implemented this form of treatment. The Maine trainees were less skeptical about family psychoeducation and more interested in receiving supervision and consultation. In Maine there was wide-ranging and local consensus before and during implementation, and more federal funding was available than in Illinois. CONCLUSIONS: Surveys of clinicians allowed prediction of which sites would successfully implement family psychoeducation services. Consensus building and funding appeared to be critical to successful outcomes. PMID- 11433112 TI - Benefits in behavioral health carve-out plans of Fortune 500 firms. AB - OBJECTIVE: This study examined the prevalence and nature of behavioral health carve-out contracts among Fortune 500 firms in 1997. METHODS: A survey was conducted of 498 companies that were listed as Fortune 500 firms in 1994 or 1995. A total of 336 firms (68 percent) responded to the survey. Univariate analyses were used to analyze prevalence, types, and amounts of covered services, cost sharing, and benefit limits. A total of 132 firms reported contracting with managed behavioral health organizations; 124 firms answered benefits questions about covered services, cost-sharing levels, and annual and lifetime limits. RESULTS: Most of the plans covered a broad range of services. Cost sharing was typically required, and for inpatient care it was often substantial. Fifteen percent of the firms offered mental health benefits that were below the limits defined in this study as minimal benefit levels, and 34 percent offered substance abuse treatment benefits that fell below minimal levels. The most generous mental health benefits and substance abuse treatment benefits, defined as no limits or a lifetime limit only of $1 million or more, were offered by 31 percent and 20 percent of the firms, respectively. CONCLUSIONS: The carve-out contracts of the Fortune 500 firms in this study typically covered a wide range of services, and the benefits appeared generous relative to those reported for other integrated and carve-out plans. However, these benefits generally did not reach the level of parity with typical medical benefits, nor did they fully protect enrollees from the risk of catastrophic expenditures. PMID- 11433113 TI - The relationship between treatment access and spending in a managed behavioral health organization. AB - OBJECTIVE: This study replicated an earlier study that showed a linear relationship between level of treatment access and behavioral health spending. The study reported here examined whether this relationship varies by important characteristics of behavioral health plans. METHODS: Access rates and total spending over a five- to seven-year period were computed for 30 behavioral health plans. Regression analysis was used to estimate the relationship between access and spending and to examine whether it varied with the characteristics of benefit plans. RESULTS: A linear relationship was found between level of treatment access and behavioral health spending. However, the relationship closely paralleled that found in the earlier study only for benefit plans with an employee assistance program linked to the managed behavioral health organization and for plans that do not allow the use of out-of-network providers. CONCLUSIONS: The results of this study replicate those of the earlier study in showing a linear relationship between access and spending, but they suggest that the magnitude of this relationship may vary according to key plan characteristics. PMID- 11433114 TI - Familiarity with and social distance from people who have serious mental illness. AB - OBJECTIVES: This study examined the effects of familiarity with and social distance from persons who have serious mental illness on stigmatizing attitudes about mental illness. METHODS: A total of 208 community college students completed three written measures about familiarity, perception of dangerousness, fear, and social distance. Path analysis with manifest-variable structural modeling techniques was used to test a version of a model in which familiarity influences the perception of dangerousness, which in turn influences fear, which influences social distance from persons with serious mental illness. RESULTS: Most of the participants reported experience with mental illness. Scores on the three written measures largely supported the path model. Correlations between the perception of dangerousness and fear as well as between fear and social distance were particularly strong. CONCLUSIONS: Approaches to social change that increase the public's familiarity with serious mental illness will decrease stigma. Further studies are warranted that focus on how contact between members of the general public and persons who have serious mental illness may be facilitated. PMID- 11433115 TI - Trauma, symptoms of posttraumatic stress disorder, and associated problems among incarcerated veterans. AB - OBJECTIVE: To help improve treatment for incarcerated veterans, the study examined exposure to trauma, symptoms of posttraumatic stress disorder (PTSD), functional status, and treatment history in a group of incarcerated veterans. METHODS: A convenience sample of 129 jailed veterans who agreed to receive outreach contact completed the Life Event History Questionnaire, the PTSD Checklist-Civilian Version (PCL-C), and the Addiction Severity Index. Participants who had scores of 50 or above on the PCL-C, designated as screening positive for PTSD, were compared with those whose scores were below 50, designated as screening negative for PTSD. RESULTS: Some 112 veterans (87 percent) reported traumatic experiences. A total of 51 veterans (39 percent) screened positive for PTSD, and 78 veterans (60 percent) screened negative. Compared with veterans who screened negative for PTSD, those who screened positive reported a greater variety of traumas; more serious current legal problems; a higher lifetime use of alcohol, cocaine, and heroin; higher recent expenditures on drugs; more psychiatric symptoms; and worse general health despite more previous psychiatric and medical treatment as well as treatment for substance abuse. CONCLUSIONS: The findings encourage the development of an improved treatment model to keep jailed veterans with PTSD from repeated incarceration. PMID- 11433116 TI - Pilot study of the effectiveness of the family-to-family education program. AB - This study assessed the efficacy of the Family-to-Family Education Program, a structured 12-week program developed by the National Alliance for the Mentally Ill. A total of 37 family members who participated in the program were evaluated by an independent research team of trained family member assessors at baseline, after completing the program, and six months after program completion. After completing the program, the participants demonstrated significantly greater family, community, and service system empowerment and reduced displeasure and worry about the family member who had a mental illness. These benefits were sustained at six months. PMID- 11433117 TI - A program for treating olanzapine-related weight gain. AB - This study evaluated the effectiveness of a Weight Watchers program for patients with schizophrenia who had olanzapine-related weight gain and ascertained whether the severity of patients' psychiatric symptoms was correlated with the patients' success in losing weight. Seven men and four women who had been treated with olanzapine and who had gained at least 7 percent of their pretreatment body weight attended Weight Watchers meetings and were offered supervised exercise sessions. The patients' weight, body mass index, and psychiatric symptoms were assessed and were compared with those of a matched comparison group who did not attend the Weight Watchers program. Only the men experienced significant weight loss. No correlation was found between weight loss and exercise or change in psychiatric symptoms. PMID- 11433118 TI - Outcomes of a cognitive-behavioral day treatment program for a heterogeneous patient group. AB - This paper describes a day treatment program that provides predominantly cognitive-behavioral therapy for a heterogeneous group of patients. Preliminary results of the program are also presented. Assessment tools included the Beck Depression Inventory, the Symptom Checklist, and a questionnaire on changes in social life. Instruments were administered at admission, at discharge, and six months after discharge. The patients showed significant improvement in scores on all instruments at discharge. Improvements were stable after six months for all diagnostic categories-depression, eating disorders, and personality disorders. The program shows promise as an effective treatment approach for patients with various psychiatric diagnoses. PMID- 11433119 TI - Forensic psychiatry services in New Zealand. PMID- 11433120 TI - The Walden House young adult HIV program. PMID- 11433121 TI - Multiple strategies of change. PMID- 11433122 TI - Incidence of restraint-related deaths. PMID- 11433123 TI - Memorizing and recalling DSM-IV diagnostic criteria. PMID- 11433125 TI - Validity of sonographic formulas for estimating fetal weight below 1,250 g: a series of 119 cases. AB - OBJECTIVE: The aim of this study was to determine the accuracy of sonographic methods for estimating fetal weight < 1,250 g on the basis of ten published formulas falling into two principal categories: general formulas applied to all fetuses, and formulas specifically developed for very-low-weight fetuses. METHODS: Recent biometric data (obtained less than 7 days before birth) on 119 fetuses weighing < 1,250 g were used retrospectively. Estimated fetal weights derived from ten published formulas were compared to actual weights. For each equation, the errors in predicting fetal weight were used to calculate mean percentage error and standard deviation of the mean error. The t test was used to determine whether the mean errors were significatively different from zero. The F test was used to determine if there were significant differences in the standard deviation of the mean errors. RESULTS: The mean birth weight of infants was 956 +/- (SD) 183 g at a gestational age of 29 +/- (SD) 2.3 weeks. The best three formulas were the Hadlock, Sabbagha and Mielke which generated a mean error of 0.25, 2.81 and 0.29 not statistically different from zero with standard deviations of 13.02, 9.14 and 11.96, which were not statistically different. CONCLUSION: In our population of very-low-birth-weight infants, the use of specific formulas targeted to those fetuses does not provide a more accurate estimation of fetal weight. PMID- 11433126 TI - Cardiovascular risk and therapeutic intervention for the early morning surge in blood pressure and heart rate. AB - The incidence of most adverse cardiovascular events appears to follow a circadian pattern, reaching a peak in the morning shortly after wakening and arising. The activities of many physiologic parameters, including hemodynamic, hematologic and humoral factors, also fluctuate in a cyclical manner over the 24h. It has been suggested that, during the post-awakening hours, the phases of these cycles synchronize to create an environment that predisposes to atherosclerotic plaque rupture and thrombosis in susceptible individuals, thereby accounting for the heightened cardiovascular risk at this time of day. Blood pressure and heart rate are part of this physiologic process, following a clear circadian rhythm characterized by a fall during sleep and a sharp rise upon awakening. This so called 'morning surge' in blood pressure may act as a trigger for cardiovascular events, including myocardial infarction and stroke. The clinical implication of these observations is that antihypertensive therapy should provide blood pressure control over the entire interval between doses. For agents taken once daily in the morning, the time of trough plasma drug level (and lowest pharmacodynamic effect) will often coincide with the early morning surge in blood pressure and heart rate. For these reasons, chronotherapeutic formulations of drugs and intrinsically long-acting antihypertensive agents provide the most logical approach to the treatment of hypertensive patients since they provide 24 h blood pressure control from a single daily dose as well as attenuating the early morning rise in blood pressure (and in some instances heart rate). PMID- 11433127 TI - Use of ambulatory blood pressure monitoring data to predict left ventricular mass in hypertension. AB - BACKGROUND: Many studies have shown definite but weak correlations between 24h blood pressure and left ventricular mass in hypertension. OBJECTIVE: The present study applied an original multivariate analysis of parameters from ambulatory blood pressure monitoring to predict left ventricular mass in hypertension. METHODS: Two hundred untreated hypertensive subjects (age=51+/-13 years, clinic blood pressure=163/98mmHg) had echocardiography and 24h recording of blood pressure. Data from 102 subjects were used to construct a mathematical model for prediction of left ventricular mass. The remaining 98 subjects were used to validate the model. RESULTS: The model included age, weight, height, 24h systolic blood pressure, heart rate over 24h, standard deviations of systolic blood pressure and of heart rate over 24h. Using this model, we explained 63% of the variance of left ventricular mass. Moreover, the model was validated for the correlation (r=0.70) between measured and the estimated left ventricular mass. CONCLUSIONS: Systolic blood pressure, heart rate and their standard deviations over 24h contribute to left ventricular mass in hypertensive subjects. Thus, it is important to take into account not only blood pressure but also heart rate in clinical studies. PMID- 11433128 TI - The influence of dose of angiotensin I-converting enzyme inhibitor on systolic blood pressure variability in heart failure: a substudy of the Assessment of Treatment with Lisinopril and Survival in heart failure (ATLAS) trial. AB - Heart failure is associated with a decreased variability in circadian systolic blood pressure. ACE inhibitors have been shown to be beneficial in CHF. However, the effect of the magnitude of the dose of ACE inhibitor on blood pressure variability has not been reported. The objective of this sub-study of the ATLAS trial was to determine if there was a difference in effect on systolic blood pressure variability of two doses (35mg, 'high'; and, 5mg, 'low') of the ACE inhibitor, lisinopril, in patients with heart failure (class II-IV; NYHA). Criteria for inclusion were: symptomatic heart failure (class II-IV; NYHA), left ventricular ejection fraction < or = 30%, and 2 months of conventional therapy with diuretics with, or without, digoxin. Twenty-four hour ambulatory blood pressure was recorded prior to randomization and after peak titration (4 weeks) of the study drug for analysis of variability of systolic blood pressure variability. The high dose of lisinopril was associated with greater variability of 24 h systolic blood pressure as noted by inspection of the 24 h recordings or calculation of the blood pressure variability index (P < 0.05). The greater variability in SBP was not associated with a difference in mean 24 h arterial blood pressure. CONCLUSIONS: Variation in circadian systolic blood pressure is useful in reflecting the influence of the magnitude of dose of the ACE inhibitor lisinopril on the pharmacodynamics of patients with heart failure. PMID- 11433129 TI - Ambulatory measurement of the QKD interval normalized to heart rate and systolic blood pressure to assess arterial distensibility--value of QKD(100-60). AB - BACKGROUND: Reduced distensibility of large arteries plays an important role in cardiovascular risk. Determination of the QKD interval during the ambulatory measurement of blood pressure enables calculation of an index of arterial distensibility. This index, the QKD(100-60), is the theoretical value of QKD at systolic blood pressure of 100mmHg and heart rate of 60bpm obtained from the linear bivariate relationship linking QKD, systolic blood pressure and heart rate on a hundred successive values measured over 24h. This study was designed to examine the relationship between QKD and QKD(100-60) on heart rate and systolic function of the left ventricle, the two parameters governing the pre-ejection time which is part of the QKD interval. METHODS AND RESULTS: In a population of 203 untreated hypertensive patients having benefited from an ambulatory measurement of blood pressure over 24h with QKD monitoring and an M-mode echocardiographic recording of the left ventricle, we found that although mean QKD was linked to heart rate and systolic function of the left ventricle, QKD(100 60) was not. It fell significantly with age, and to a greater extent in the sustained hypertensives than in white-coat hypertensives. CONCLUSION: QKD(100-60) constitutes an index of arterial distensibility independent of the pre-ejection time. As an adjunct to the ambulatory measurement of blood pressure, its determination is simple and completely automatic, thus eliminating observer bias. PMID- 11433130 TI - Canadian valsartan study in patients with mild-to-moderate hypertension. AB - OBJECTIVE: This study was designed mainly to establish the rates of response to valsartan 80mg once daily (qd) and to valsartan 160mg qd given to non-responders to 80mg qd, as well as to determine the safety of valsartan and the blood pressure control achieved using valsartan over a period of 24 h or more, using ambulatory blood pressure monitoring (ABPM) devices. METHODS: This was a single blind, single-arm, multicenter study involving 256 out-patients with mild-to moderate essential hypertension. After previous antihypertensive treatments had been 'washed out', the patients were entered into a 2-week placebo run-in period to confirm the diagnosis of mild-to-moderate hypertension. Patients who, at the end of the placebo run-in period, had a mean sitting diastolic blood pressure of between 95 and 115mmHg inclusive received valsartan 80mg qd for 4 weeks. Non responders (those not demonstrating a diastolic blood pressure of less than 90mmHg or a decrease in diastolic blood pressure of 10mmHg or more compared with baseline) received valsartan 160mg qd for another 4 weeks. In selected centers, patients who agreed also had their blood pressure monitored for 24 h, provided their blood pressure was shown to be controlled. Of these patients, half skipped one dose of valsartan and were monitored for an additional 24h period. RESULTS: The rate of response to valsartan 80mg was 45.4%, and of those not responding to this dose, 36.3% responded to valsartan 160mg. The response rate to one or other dose was 63.2%. The ambulatory blood pressure data support a consistent reduction of blood pressure with valsartan over a 24h period and for up to 32 h after dosing in those who missed a dose. The overall incidence of adverse experiences per person-year, treatment related or otherwise, was 6.3 and 10.6 for the valsartan and placebo study periods respectively. CONCLUSION: Antihypertensive treatment with valsartan for 8 weeks produced a significant decrease in diastolic blood pressure in hypertensive patients. In addition, the drug may be safely administered, and the results of 24 h/48 h ambulatory monitoring demonstrate that valsartan is a true once-a-day antihypertensive. PMID- 11433131 TI - Accuracy of aneroid sphygmomanometers in clinical usage: University of Michigan experience. AB - BACKGROUND: Aneroid manometers are frequently used to measure blood pressure. Aneroid manometers have moving parts that are subject to fatigue. The accuracy duration of the aneroid devices, like most digital devices, is unstudied. It has been accepted that if the aneroid device does not rest at '0' it is inaccurate, but how often is the device inaccurate when it does rest at '0'? METHODS: A Universal Biometer DPM-III measuring unit was used for all of the measurements at 10 University of Michigan Health System sites. A total of 136 aneroid manometers were tested. Two additional aneroid devices were not tested, as the needle did not start within '0'. Static pressure measurements were made at nine levels for all devices: 50, 80, 90, 100, 120, 150, 200 and 250mmHg. RESULTS: The average difference of the nine pressure settings of the whole group was 0.2+/-0.31 (95% confidence interval 0.1-0.2) mmHg. The largest number of devices that were not calibrated within +/-3mmHg was seen at the 150mmHg setting with six (4.4%) of the devices failing. If an accuracy standard of +/-2mmHg was used, the largest number of devices failed at 250mmHg (22 devices, 16.2%). The largest number of devices that were inaccurate was manufactured 6 years prior to testing and was from two sites. CONCLUSIONS: Aneroid devices were accurate. A yearly calibration programme should be performed and a +/-2mmHg standard should be used. Portable aneroid manometers may need to be more frequently calibrated due to the trauma associated with dropping. PMID- 11433132 TI - Evaluation of the overall efficacy of the Omron office digital blood pressure HEM 907 monitor in adults. AB - BACKGROUND: Non-invasive self blood pressure monitoring has become increasingly popular. To assure the accuracy of devices used for this purpose, all need to be validated independently prior to marketing. The objective of this study was to assess the accuracy of the HEM-907, a new semi-automatic, non-invasive, oscillometric blood pressure monitoring device specifically designed to be used in the clinic or physician's office setting. METHODS: Blood pressure measurements taken employing this device were compared with the results obtained by two experienced observers using a mercury sphygmomanometer on 100 subjects and patients (384 measurements). The limits of agreement were calculated for the device compared with the results of the two observers according to the standards of the Association for the Advancement of Medical Instrumentation (AAMI). RESULTS: The agreement between the two observers was -0.36+/-2.32mmHg for systolic blood pressure and 0.02+/-2.42mmHg for diastolic blood pressure. The agreement between the device and the observers was 1.56+/-4.42mmHg and 3.49+/ 4.61mmHg for systolic and diastolic blood pressure respectively. CONCLUSIONS: The Omron HEM-907 satisfied the AAMI criteria for accuracy for a non-invasive blood pressure monitoring device. PMID- 11433134 TI - The long-term functional outcome of operatively treated tibial plateau fractures. AB - OBJECTIVES: To review the long-term functional results of the surgical treatment of tibial plateau fractures using standard techniques of open reduction and internal fixation. DESIGN: Retrospective study. SETTING: University hospital. METHODS: Forty-seven displaced fractures of the tibial plateau in forty-six patients were treated with open reduction, interfragmental screw fixation of the articular fragments, and buttress plate fixation and had a minimum of five years of follow-up. All aspects of their care, including tibial plateau fracture type, operative management and associated injuries, were documented. Preoperative and postoperative follow-up radiographs were analyzed for fracture classification and adequacy of reduction. All patients were contacted and given functional outcome questionnaires using both a generic health status scale (Short Form 36 [SF-36]) (18) and a disability scale relating to knee osteoarthritis (Western Ontario and McMaster Universities Osteoarthritis index [WOMAC]) (1). Data were also collected regarding return to work and sporting activities. Assessment scores were analyzed with respect to age, fracture type and severity, and were compared to standardized age and sex-matched scores for the healthy population. The average age of the patients at the time of injury was forty years and the average follow up period was 8.3 years. Of the forty-seven fractures studied, twenty-five were classified as Schatzker types I, II, or III, and the remaining twenty-two were types IV, V, or VI (15). All fractures received operative treatment within forty eight hours and all but five fractures were acceptably reduced. RESULTS: Compared to the standardized SF-36 categorical and aggregate scores, there was no statistically significant difference between the healthy age-matched population and twenty-four of twenty-six of the under-age-forty group regardless of fracture type. With regard to the over-age-forty group, scores statistically similar to the control population were found in only twelve of twenty-one patients. Although there was a large variance in WOMAC scores for all groups resulting in no statistically significant difference being found, a trend toward higher categorical and aggregate scores was seen with increasing age at presentation. There was no correlation between WOMAC scores and fracture type. Multiple classifications analysis of all data revealed that presentation age was the most significant source of variation with respect to functional outcome. Fracture type had much less influence and adequacy of reduction had no significant influence on outcome, although the group of patients having an inadequate reduction by the authors' criteria was too small in number to reasonably comment upon. CONCLUSIONS: Open reduction and internal fixation is a satisfactory technique for the treatment of displaced fractures of the tibial plateau, particularly for patients younger than forty years. PMID- 11433135 TI - Cortisol kinetics in elderly women with persistently high cortisol concentrations after proximal femur fracture. AB - OBJECTIVES: Elderly women with proximal femur fracture show abnormal persistence of increased cortisol concentrations, which could contribute to the high morbidity associated with this injury. Two weeks after injury, the authors found substantially increased urinary free cortisol excretion, which usually reflects the integrated concentration of free (bioactive) cortisol in plasma. However, there was a proportionally smaller increase in cortisol production rate. The authors have now tested the hypothesis that this was caused by a decreased metabolic clearance rate (MCR) rather than increased renal clearance, because the latter but not the former would invalidate free cortisol excretion as an index. SETTING: Orthopaedic wards in a teaching hospital. PATIENTS: Thirteen women aged seventy-one to ninety-two years who had sustained a proximal femur fracture approximately two weeks earlier were compared with ten healthy women aged sixty seven to eighty-three years. These subjects are similar to those in the authors' previous study. MAIN OUTCOME MEASUREMENTS: The authors used single injections of [3H] cortisol to measure its MCR and estimated hepatic blood flow with indocyanine green. RESULTS: The patients with hip fractures had higher plasma cortisol concentrations than did the healthy subjects, as expected. Cortisol MCR was approximately 20 percent lower in the patients, and estimated hepatic blood flow was approximately 35 percent lower in the patients. Analysis of covariance showed that the difference in MCR was the result of the small difference in age between the groups rather than to injury per se. CONCLUSIONS: A lower cortisol MCR in the patients with hip fractures explains the authors' previous results and validates urinary free cortisol excretion as an index. The data suggest a roughly threefold mean increase in plasma cortisol bioactivity two weeks after hip fracture. PMID- 11433136 TI - A preliminary study of joint surface changes after an intraarticular fracture: a sheep model of a tibia fracture with weight bearing after internal fixation. AB - OBJECTIVE: To evaluate the changes in the articular cartilage and subchondral bone after an osteotomy designed to simulate an articular fracture. DESIGN: The contribution of the cartilage and subchondral bone was evaluated twelve weeks after creating a 1.0-millimeter step-off in the medial plateau of the tibia of twelve adult domestic sheep. All animals surviving were labeled with fluorescent markers for bone production, oxytetracycline (fifty milligrams per kilogram), and calcein (twelve milligrams per kilogram) nine and 11.5 weeks after surgery. The knees were loaded in compression using an Instron materials tester with pressure sensitive film to record joint contact pressures above and below the medial meniscus. SETTING: The studies were performed in the research laboratories of the Orthopaedic Laboratory at Harborview Medical Center in Seattle, Washington and the Madigan Army Medical Center in Tacoma, Washington. ANIMALS: Twelve adult domestic sheep. INTERVENTION: An intraarticular osteotomy of the medial tibial plateau with 1.0 millimeter of displacement was performed. The osteotomy was stabilized with 3.5-millimeter lag screws. MAIN OUTCOME MEASURES: The contact pressures of the knee joint and articular histology were evaluated twelve weeks after surgery. Samples of the articular cartilage were analyzed by light microscopy and electron microscopy to evaluate the response of the articular cartilage and subchondral bone of the differential joint loading because of the irregularity in the articular surface caused by the osteotomy. RESULTS: The knees with an intraarticular step-off had two major contact areas with an intervening zone of reduced load corresponding to the edge of the depressed fragment. Coronal histologic sections through the articular surface showed the presence of thinning and fibrillation on the high side of the step-off and some compensatory hypertrophy of the cartilage. The subchondral bone was not responsible for restoring articular congruity because the rate of bone production was similar between the low side of the articular fracture (1.85 micrometers per day) and the high side of the fracture (1.67 micrometers per day). Scanning electron microscopy showed partial cartilage remodeling by deformation of the high side cartilage with bending of the vertical collagen fibrils, even in the unloaded state. CONCLUSIONS: In this model with a small fracture displacement (1.0 millimeter), which was less than the thickness of the articular cartilage (1.5 millimeters), the contour of the joint improved despite residual articular surface incongruency after the fracture healing. PMID- 11433137 TI - Screw position affects dynamic compression plate strain in an in vitro fracture model. AB - OBJECTIVE: This investigation considers the effect of a variety of screw positions on plate strain in three fracture models. DESIGN: Dynamic compression plate fixation of in vitro fracture models. METHODS: To model a fracture, a plastic pipe was cut transversely and a twenty-hole dynamic compression plate was attached by screws. Eighteen stacked, rectangular, rosette strain gauges were installed on the plate to evaluate strain. Three models were evaluated: two constructs in which there was no contact between the cut ends of the pipe under the fixation plate (small-and large-gap models) and a construct in which there was direct apposition of the cut ends (no-gap model). The pattern and magnitude of strains were assessed as a function of varying combinations of screw position for each model. RESULTS: Maximal plate strain in the gap models was lowest with screws placed closest to the gap, compared with screws placed away from the gap or spaced apart. The no-gap model showed significantly lower strains when screws were placed further from the osteotomy site than when screws were positioned close together or spaced apart. In all cases, maximal plate strain occurred adjacent to the most central screw holes and rapidly dissipated along the length of the plate. CONCLUSION: In a model simulating a comminuted fracture (gap), this study found that screws should be placed as close to the fracture site as possible to minimize plate strain. In an anatomically reduced two-part fracture model (no gap), widely spaced screws or those placed away from the fracture resulted in lower strains. PMID- 11433138 TI - Factors predictive of immobilization complications in pediatric polytrauma. AB - OBJECTIVES: To determine in a cohort of children with polytrauma which variables are predictive of the development of complications related to immobilization. DESIGN: A retrospective study of children with polytrauma and at least one major musculoskeletal injury. A stepwise forward logistic regression analysis was used to determine variables predictive of complications related to immobilization. PARTICIPANTS: Ninety-three children with polytrauma were studied; motor vehicle incidents accounted for 80 percent of the injuries. The average age was 8.0 +/- 4.1 years. There were 152 fractures in the ninety-three children. The average Modified Injury Severity Scale (MISS) was 24.5 +/- 13.6. There were thirty-five complications in twenty-two children, and four children died. RESULTS: Two variables were predictive of complications related to immobilization: age and MISS score. Complications related to immobilization were positively associated with being older than seven years of age (p = 0.027; odds ratio = 9.5; 95 percent confidence interval 1.4, 64.9) and having a MISS score greater than forty (p = 0.005; OR = 14.1; 95 percent confidence interval 2.2, 89.1). Timing of surgery showed a trend (p = 0.097) but did not reach statistical significance. CONCLUSIONS: Complications of immobilization in children with polytrauma are associated with age greater than seven years and a MISS score greater than forty. Further study is needed to evaluate the effect of early fracture stabilization. Timing of osteosynthesis showed a trend but did not reach statistical significance in this study. PMID- 11433139 TI - Clinical performance of a new medullary humeral nail: antegrade versus retrograde insertion. AB - OBJECTIVE: A new intramedullary nail system for humeral shaft fractures is evaluated to determine whether retrograde nailing is as reliable as antegrade nailing. STUDY DESIGN: Prospective multicenter nonrandomized clinical study. PATIENTS: Eighty-four patients with acute humeral shaft fractures were nailed with the new unreamed humeral nail (UHN) system. Fifty-seven nails were introduced retrogradely, and twenty-seven antegradely. Bone healing and functional outcome were the follow-up parameters. RESULTS: The ratio of perioperative complications was equivalent for both groups, but one shaft fracture and three fractures or fissures at the entry point occurred in the group with retrograde nail insertion. Five fractures, all with retrograde nail insertion, needed secondary surgery to achieve bony healing. There was no difference in functional outcome after healing in either group. CONCLUSION: Retrograde nailing of humeral shaft fractures is technically more demanding than antegrade nailing. Fractures or fissures at the insertion point must be avoided by adequate preparation of the entry hole and careful nail insertion. Bone healing problems seem more surgeon-related than approach-related. As in every other procedure, an optimal fracture configuration and high fracture stability must be achieved. PMID- 11433140 TI - Open reduction and plate fixation of displaced AO type C3 fractures of the distal radius: restoration of articular congruity in eighteen cases. AB - OBJECTIVE: To determine the ability of open reduction and plate fixation to restore articular congruity in the treatment of fractures of the distal radius with intraarticular comminution and displacement. DESIGN: Prospectively followed series. METHODS: Eighteen consecutive fractures of the distal radius with intraarticular comminution and displacement were treated with open reduction and combined volar and dorsal plate fixation in thirteen, dorsal plate fixation in four, and volar plate fixation in one case. Kirschner wires were added in thirteen fractures and cancellous bone graft was used in four fractures. Articular congruity after a mean follow-up of twenty-three months was assessed using anteroposterior and lateral radiographs. RESULTS: Articular congruity without a two millimeter or more intraarticular stepoff was found at follow-up in fifteen of eighteen cases, without a two millimeter or more gap in nine cases and without a two millimeter or more stepoff or gap in only eight cases. One reason for not having obtained articular congruity in some of the fractures was insufficient intraoperative visualization of the joint surface. Three reduced fractures redisplaced. The reason for loss of intraarticular reduction was considered insufficient stabilization of the distal fragments. In two of the three cases, it was thought that insertion of bone graft would have prevented the loss of reduction. CONCLUSIONS: Articular incongruity after open reduction and plate fixation of comminuted fractures of the distal radius may occur more often than expected. The reduced fracture should be evaluated by intraoperative radiographs. If the reduced joint surface is not entirely visible on the radiographs because of implants or other reasons, oblique radiographs or fluoroscopy should be additionally used. Plate fixation alone does not always provide sufficient stability and cancellous bone graft should be added in cases of metaphyseal bone loss or for stabilization of multiple small articular fragments that cannot be fixed by screws or Kirschner wires. PMID- 11433141 TI - Pauwels' classification of femoral neck fractures: correct interpretation of the original. AB - SUMMARY: Pauwels' classification has been in use as a therapeutic guideline since 1935. Although its application is currently limited, it is still referred to and frequently misinterpreted by authors. The causes of this misinterpretation relate first of all to the degrees of the inclination of the fracture line in Degrees II and III of the classification. The correct values of Pauwels' classification are the following: Degree I, up to 30 degrees; Degree II, between 30 and 50 degrees; and Degree III, 50 degrees and more. PMID- 11433142 TI - Intraarticular entrapment of the obturator nerve in acetabular fracture. AB - SUMMARY: The authors report a case of acetabular fracture of both columns that was associated with a rare nerve injury, entrapment of the obturator nerve in the hip joint. PMID- 11433143 TI - Operative treatment of a transverse fracture-dislocation at the S1-S2 level. AB - SUMMARY: The authors recently treated a multiply injured patient who sustained a rare fracture-dislocation at the S1-S2 level. The S1 vertebra was displaced forward into the pelvic cavity and was located just in front of the S2 vertebra. Because the patient also had extensive neurologic injury to the lumbar plexus and instability of the pelvic ring, operative treatment was deemed necessary. Surgery to stabilize the pelvis and decompress the lumbar plexus proved successful, and the patient experienced marked improvement in her postoperative neurologic function. Nonoperative treatment has traditionally been recommended for this injury, but advances in spinal surgery have made transverse sacral fractures more amenable to open reduction and fixation. Potential benefits of this operative treatment include relief of pressure from the lumbar plexus, a stable pelvis and facilitation of return of neurologic function. PMID- 11433144 TI - Bilateral anterior and posterior traumatic hip dislocation. AB - SUMMARY: A case report of simultaneous traumatic bilateral anterior and posterior hip dislocations is presented, and the management of this injury is discussed. PMID- 11433145 TI - Open reduction internal fixation after displacement of a previously nondisplaced acromial fracture in a multiply injured patient: case report and review of literature. AB - SUMMARY: A twenty-eight-year-old multiple trauma patient had a nondisplaced acromion fracture that was not detected until after it had displaced. Open reduction internal fixation was performed without complication and the patient achieved excellent shoulder abduction strength. Nondisplaced acromion fractures may displace if not protected. Open reduction internal fixation of displaced acromion fractures should be considered if deltoid muscle strength is important to the patient. PMID- 11433146 TI - Type I Monteggia fracture dislocation associated with ipsilateral distal radial epiphyseal injury. AB - SUMMARY: Ipsilateral elbow and wrist injuries are rare in children. We present a previously undescribed case of a Bado Type I Monteggia fracture with a Salter Harris Type II epiphyseal injury of the distal radius. A satisfactory result was achieved but required surgical intervention. Complete diagnostic evaluation is imperative to avoid missed injuries and achieve satisfactory outcome. PMID- 11433147 TI - Three epiphyseal fractures (distal radius and ulna and proximal radius) and a diaphyseal ulnar fracture in a seven-year-old child's forearm. AB - SUMMARY: The authors report a rare case of fracture separations at both ends of the radius combined with an epiphyseal and diaphyseal fracture of the ipsilateral ulna. A seven-year-old girl fell one story and sustained a closed injury of her forearm. A closed reduction was unsuccessful, and an open reduction was performed with three of the four fractures being secured with Kirschner wires. These wires were removed one month later, and range-of-motion exercises were started. Thirty months after surgery, both forearms were equal in length, although the proximal radial epiphyseal line appeared partially closed. Joint motions, including forearm rotation, were normal. Radiologically, the ulnar diaphysis and the radial neck were posteriorly convex 20 degrees and 18 degrees, respectively. PMID- 11433148 TI - Re.: Acute noncontact compartment syndrome. PMID- 11433149 TI - The cavus foot. PMID- 11433150 TI - Comparison of three outcomes instruments in children. AB - SUMMARY: The purpose of this study was to compare prospectively the distributions, validity, and discriminative ability of three pediatric outcome questionnaires. Consecutive patients completed the Activities Scales for Kids (ASK), the Child Health Questionnaire Parent Form (CHQ-PF-28), and the Pediatrics Outcomes Data Collection Instrument (PODCI). The scores of the three instruments were compared with each other and with parents' and clinicians' ratings. Of 210 patients, 166 (79%) completed the three questionnaires. The CHQ-PF-28 had a different distribution than the other two questionnaires and showed both floor and ceiling effects. The ASK and PODCI instruments were highly correlated and discriminated better than the CHQ-PF-28, with fewer floor and ceiling effects. Each questionnaire, however, seemed to be measuring slightly different things. PMID- 11433151 TI - Evaluation of the efficacy of pelvic shielding in preadolescent girls. AB - SUMMARY: A standing anteroposterior pelvic radiograph with gonadal shielding is used as a screening tool for all patients evaluated for intoeing at our institution. Sixty-two normal consecutive screening pelvic radiographs obtained in 61 female patients between the ages of 4 and 6 years were evaluated. Radiographs were evaluated for the adequacy to assess the hips as well as the protection afforded the ovaries from radiation exposure. Radiographs were judged to be inadequate because the shield covered essential landmarks in at least one hip in eight radiographs (13%). Five radiographs (8%) covered >50% of the area of both ovaries, and only one radiograph covered >75% of the area of both ovaries. Standard techniques of positioning gonadal shields in preadolescent girls are inadequate and provide minimal protection with a high rate of interference with vital landmarks. We no longer advocate using gonadal shields on initial screening radiographs of preadolescent girls. PMID- 11433152 TI - Treatment of femoral fractures in children by pediatric orthopedists: results of a 1998 survey. AB - SUMMARY: This study aimed to determine treatment preference of various femoral fracture patterns in children by pediatric orthopedists and whether it is practice dependent. In September 1998, members of the Pediatric Orthopedic Society of North America were surveyed to determine their current preferences in treating each of four middle one-third femoral fracture patterns in four age groups. Forty-four percent (286/656) of those surveyed responded. For each fracture pattern, operative treatment was increasingly preferred over nonoperative as patient age increased, and the preferred treatments within the operative and nonoperative categories changed significantly as patient age increased. Fourteen specific cases of femoral head avascular necrosis were noted after rigid reamed and unreamed rodding. There is a statistically significant trend by pediatric orthopedists to treat older children's femur fractures operatively and younger children's nonoperatively. The consensus treatment is age dependent. The numerous cases of avascular necrosis after rigid rodding are a concern. PMID- 11433153 TI - Ender nail fixation of pediatric femur fractures: a biomechanical analysis. AB - SUMMARY: Biomechanical testing was performed to determine the effects of flexible intramedullary nail fixation on simulated transverse and comminuted midshaft femur fractures using two Ender nails. A synthetic adolescent-size femur model was used. The axial and rotational stiffness values for the simulated comminuted fracture were equivalent to those of the transverse fracture under "touch-down weight-bearing" loads. These data suggest that length and rotational control of comminuted midshaft femur fractures with two divergent Ender nails may be sufficient for early mobilization. PMID- 11433154 TI - Analysis of the cause, classification, and associated injuries of 166 consecutive pediatric pelvic fractures. AB - SUMMARY: Pediatric pelvic fractures are serious injuries. Anatomical differences exist between pediatric and adult populations, leading to different causes and rates of death, fracture patterns, and associated injuries. This study is the largest consecutive series of pediatric pelvic fractures from one institution emphasizing the unique aspects seen in pediatrics. One hundred sixty-six children were included. Plain radiography and computed tomography scans were used to classify pelvic fractures. Multisystem injuries occurred in 60%, and 50% sustained additional skeletal injuries. The death rate was 3.6%. Head and/or visceral injuries were the causes of all deaths. Life-threatening hemorrhage did not occur. Urethral injury was not seen as often as in adults. Anterior ring fractures were the most common type, dominated by pedestrian versus motor vehicle trauma. Anatomical differences and mechanism of injury may play a role in these contrasting findings. PMID- 11433155 TI - Coronal split fracture of the proximal tibia epiphysis through a partially closed physis: a new fracture pattern. AB - SUMMARY: A comminuted coronal split fracture of the proximal tibial epiphysis is an uncommon injury in children. The authors evaluated and treated two patients who sustained an epiphyseal fracture through a partially closed proximal tibial epiphysis. Plain radiography and computed tomography with three-dimensional reconstruction showed a comminuted coronal split fracture of the proximal tibial epiphysis. Each patient underwent definitive operative fixation of the fracture and was followed at least 1 year after clinical union. A mechanism consisting of three-point bending on the tibial plateau is proposed. PMID- 11433156 TI - Pediatric floating elbow. AB - SUMMARY: A retrospective review of 16 patients with floating elbow injuries over a 9-year period at a tertiary care children's hospital confirms that these injuries are associated with substantial swelling and the potential to develop compartment syndrome, particularly when circumferential cast immobilization is used. Among 10 patients in whom the forearm was treated with closed reduction and plaster immobilization, a compartment syndrome developed in 2, and 4 patients had incipient compartment syndrome that responded to splitting of the cast; 3 of these subsequently required remanipulation of the distal radius. One patient with compartment syndrome had Volkmann ischemic contracture. Six patients underwent stabilization of both the distal humeral and forearm fractures with percutaneously inserted Kirschner wires, thereby allowing postreduction immobilization in a split cast. None of these patients had problems with excessive swelling or compartment syndrome. Percutaneous Kirschner wire fixation of both the humeral and forearm fractures in pediatric floating elbow injuries allows noncircumferential immobilization, thereby reducing the risk of compartment syndrome. PMID- 11433157 TI - Flexion-type supracondylar elbow fractures in children. AB - SUMMARY: Flexion-type supracondylar elbow fractures are uncommon in children. Minimally displaced fractures were treated in plaster cast in 7 children, and 22 children with a displaced fracture were treated by closed reduction and percutaneous pinning. Results after an average follow-up of 6.3 years showed that the treatment of choice yielded excellent or good results in 86.2% of the patients. All patients were satisfied with the end result and had normal use of their elbow. PMID- 11433158 TI - Normal ranges of scapholunate distance in children 6 to 14 years old. AB - SUMMARY: The purpose of this study was to determine the normal ranges of scapholunate distance (SLD) applicable to children. Bone age radiographs of 85 children (36 boys, 49 girls) 6 to 14 years of age were used to establish the normal ranges. Repeated-measures analysis of variance revealed significant age and gender differences. Therefore, linear regression was used to determine normal SLD ranges separately for girls and boys based on chronologic age and bone age. SLD normal ranges were similar for chronologic age and bone age. Interobserver and intraobserver agreement was excellent. There was no significant age by gender interaction found, indicating that although boys had significantly longer distances than girls, both showed a similar rate of decreasing SLD distance with age. These results provide age-and gender-based normal ranges of SLD in young children. These results may be useful in assessing traumatic soft tissue injuries about the pediatric wrist. Of note, only three girls and two boys (all 12 years or older) met the adult criterion of normal SLD of < or = 2 mm. PMID- 11433159 TI - Use of allografts in Pemberton osteotomies. AB - SUMMARY: Twenty patients treated with 26 Pemberton osteotomies with allograft bone wedges were retrospectively evaluated radiographically. Two patients undergoing four osteotomies were excluded because of incomplete records or <1 year of follow-up. Eleven of the remaining 18 patients had other procedures performed. Osteotomies all united radiographically within 3 months. The acetabular index improved from 33 degrees before surgery to 18 degrees after surgery, and the lateral migration index improved from 60% to 26%. This new technique of Pemberton osteotomy with patellar allograft wedges allows good correction of acetabular dysplasia with immediate graft stability. This stability often obviates the need for postoperative spica casting, even for patients undergoing bilateral procedures, thereby allowing more rapid rehabilitation. PMID- 11433160 TI - Long-term follow-up of congenital subluxation of the hip. AB - SUMMARY: This is a retrospective study of long-term radiographic results for congenital subluxation of the hip (CSH) after closed reduction at the authors' hospital from 1963 to 1980. The age at final follow-up ranged from 14.0 to 33.9 years (average 19.1). The diagnosis of CSH was determined radiographically using Ishida criteria. The center-edge angle (CE angle) and the Sharp angle were measured serially, and the final results were evaluated using Severin classification, and evaluation of avascular necrosis used the classification of Kalamchi and MacEwen. Sixty-nine (26.3%) of 262 hips with CSH were rated as Severin group III or IV. Avascular necrosis was found in 12 hips (4.6%). The CE angle of unaffected hips had strong correlation with that of affected hips. These late results were not satisfactory and indicate that patients with CSH should be followed up at least until skeletal maturity, as is done with complete hip dislocation. PMID- 11433161 TI - Clinical characteristics of children with atypical and idiopathic slipped capital femoral epiphysis: description of the age-weight test and implications for further diagnostic investigation. AB - SUMMARY: Slipped capital femoral epiphyses (SCFEs) can be idiopathic or atypical (associated with renal failure, radiation therapy, and endocrine disorders). The demographics of 433 children (285 idiopathic, 148 atypical) with 612 SCFEs were studied to define predictors of atypical SCFEs. Multiple logistic regression analysis showed that age and weight were predictors. For two patients of equal weight, those younger than 10 or older than 16 years of age were 4.2 times more likely to have an atypical SCFE; for two patients of equal age, those <50th percentile weight were 8.4 times more likely. The age-weight test was defined as negative when age younger than 16 years and weight > or = 50th percentile and positive when beyond these boundaries. The probability of a child with a negative test result having an idiopathic SCFE was 93%, and the probability of a child with a positive test result having an atypical SCFE was 52%. An evaluation of the child's age and weight is useful when considering the cause of an SCFE. PMID- 11433162 TI - Effect of femur position on the angular measurement of slipped capital femoral epiphysis. AB - SUMMARY: A femur model was used to simulate a slipped capital femoral epiphysis (SCFE). Wedges were inserted into a cut in the femoral neck to create SCFEs of 25, 50, and 75 degrees. The effects of femur position on the angular measurement of SCFE were studied by taking radiographs of the SCFE at preselected positions of rotation and abduction, which were ensured by a holding device. The views selected were the cross table lateral (CTL), Billing lateral, and frog lateral (FL) projections. FL views were obtained at 22.5, 45, and 67.5 degrees of abduction. The rotational positions were 20 degrees internal, neutral, and 15, 30, 45, and 60 degrees external. Slip angles were measured from these radiographs. The CTL and FL projections accurately portrayed any severity of an SCFE at all positions except for 45 and 60 degrees of external rotation. The Billing view falsely portrayed a severe SCFE at all positions of rotation. The most accurate representation of an SCFE is seen on either the CTL or FL view when the external rotation of the femur does not exceed 30 degrees. PMID- 11433163 TI - Value of measurement of hip movements in childhood hip disorders. AB - SUMMARY: After the reproducibility of measurement of hip joint motion by different techniques was ascertained, the ranges of motion of hips of 325 normal schoolchildren ages 5 to 14 years were measured with a fluid level goniometer. All movements of the hip decreased in a nonlinear fashion with increasing age. The regression line with the closest fit to the mean values of each hip movement was that obtained by applying the sixth-order equation. A grid was designed to record the ranges of movement of the hip. The ranges of movement of 93 children with pathology in one or both hips were measured by the same technique and marked on the grid. Characteristic abnormal grid patterns were noted for each condition. Careful documentation of alterations in hip joint movement in children with symptoms related to the hip can be of value in establishing the diagnosis of the underlying pathology. PMID- 11433164 TI - Synostosis of the ring-small finger metacarpal in Apert acrosyndactyly hands: incidence and treatment. AB - SUMMARY: This study reports on the incidence of a synostosis between the ring small metacarpal bases in patients with Apert syndrome and describes a technique to resect the synostosis and insert silicone sheets to improve hand function. Records of 9 patients (18 hands) were evaluated. Average age at follow-up was 9.5 years. Three relationships between the ring-small metacarpals were observed. Type I hands (22%) had no abnormal interconnections; type II hands (33%) possessed a synostosis since birth; type III hands (44%) did not possess a synostosis at birth, but one later developed at average age of 79 months. Total incidence of synostosis was 77%. Five patients underwent excision of the synostosis. At an average of 16 postoperative months, 4 of these patients experienced improvement in hand prehension. In patients with Apert syndrome, resection of abnormal interconnections between the ring-small metacarpals is highly recommended to improve hand prehension based on the high incidence of a synostosis in this region. PMID- 11433165 TI - Vitamin d deficiency in breast-fed toddlers. AB - SUMMARY: The evaluation of genu varum and intoeing in young toddlers is a frequent problem seen by the primary care physician and the orthopaedic surgeon. This report describes six nonwhite breast-fed toddlers with extreme genu varum. Clinical and radiographic findings were consistent with vitamin D-deficiency rickets. Within 3 to 6 months of the initiation of ergocalciferol treatment, reparative processes were evident on both radiographic and clinical examinations. Laboratory studies also showed underlying correction. With the current emphasis on the benefits of breast-feeding and the limitation of sunlight, this diagnosis must remain on the differential in the evaluation of genu varum. PMID- 11433166 TI - Musculoskeletal deformities in Behr syndrome. AB - SUMMARY: Seventeen children with Behr syndrome were investigated, focusing on the musculoskeletal deformities and long-term outcome. Behr syndrome is characterized by optic atrophy beginning in early childhood associated with ataxia, spasticity, mental retardation, and posterior column sensory loss. The ataxia, spasticity, and muscle contractures, mainly of the hip adductors, hamstrings, and soleus, are progressive and become more prominent in the second decade. In 70% of the patients, contractures developed in the lower limbs, requiring surgery mainly for the Achilles tendon, hamstrings, and adductor longus. At last follow-up at an average age of 21.7 years (range, 8-31 years), 13 of the patients are housebound walkers, 2 are nonfunctional walkers, and 2 are nonwalkers. PMID- 11433167 TI - Rod diameter prediction in patients with osteogenesis imperfecta undergoing primary osteotomy. AB - SUMMARY: Because the cross-sectional shape of the long bones of patients with osteogenesis imperfecta is often elliptical, the use of preoperative radiographs to determine intramedullary rod diameter in these patients undergoing osteotomy may be misleading. To investigate this, the authors correlated the narrowest inner bone diameter (NID) on preoperative radiographs to the rod diameter (RD) on postoperative radiographs. The authors evaluated 79 bones in 27 patients undergoing primary osteotomy with intramedullary fixation. Only 5% of the bones had an equal NID and RD, with 81% of bones having a smaller RD than the measured NID. Although a positive correlation was found between RD and NID (correlation coefficient 0.76), measurement of the NID on preoperative radiographs did not provide a good prediction of the actual RD used in this series of children with osteogenesis imperfecta. PMID- 11433168 TI - Efficacy of perioperative halo-gravity traction in the treatment of severe scoliosis in children. AB - SUMMARY: Perioperative halo traction was used in the treatment of severe scoliosis in 19 children. Diagnoses included neuromuscular, idiopathic, and congenital scoliosis. Traction was transferable between the bed and a walker or wheelchair. Thirteen patients had prior spinal surgery, and most required osteotomy. Traction was used for 6 to 21 weeks. All patients underwent spinal fusion surgery after traction, with instrumentation used in 15 patients. Improvement was achieved in all patients. The Cobb angle improved 35% from an average 84 degrees before traction (range 63 degrees -100 degrees ) to 55 degrees preceding fusion. Trunk decompensation improved in all patients. Trunk height increased 5.3 cm in traction. Response to traction did not correlate with diagnosis, patient age, or prior surgery. There were no neurologic complications. Perioperative halo-gravity traction improves trunk balance and frontal and sagittal alignment in children with severe spinal deformity. Surgical fusion was enhanced by the improved alignment, and neurologic injury was avoided. PMID- 11433169 TI - Evaluating congenital spine deformities for intraspinal anomalies with magnetic resonance imaging. AB - SUMMARY: The incidence of intraspinal abnormalities associated with congenital spinal anomalies as detected by magnetic resonance imaging (MRI) is becoming better defined. In this study, 41 nonrandomized children with congenital spinal deformities (excluding myelomeningocele) who underwent complete MR evaluation were reviewed. Of the 41 congenital spinal deformities, 37 demonstrated congenital scoliosis, with failure of formation in 19, failure of segmentation in 4, and mixed defects in 14. The remaining four deformities were cases of congenital kyphosis. Thirteen patients with congenital spine anomalies were noted to have intraspinal abnormalities identified by MRI: tethered cord in 12 patients, syringomyelia in 3 patients, and diastematomyelia in 5 patients. Of the 12 patients with tethered cord, 2 patients had neurologic deficits. Urorectal anomaly was one of the most common associated findings (15%). Considering an incidence of intraspinal anomalies of 31% and as clinical manifestations may not be initially detectable, MRI is recommended in patients with congenital spinal deformity as part of the initial evaluation even in the absence of clinical findings. PMID- 11433170 TI - Cerebral microembolization during pediatric scoliosis surgery: a transcranial doppler study. AB - SUMMARY: The goal of this study was to identify cerebral microemboli during scoliosis surgery and their potential relationship with visual alterations. Transcranial Doppler identified high-intensity transient signals (HITS) during surgery in both middle cerebral arteries, and ophthalmologic examination assessed their potential effects on the visual system. Thirteen children (age 13-17 years) undergoing surgery for scoliosis or kyphosis with spine curvature >45 degrees were studied. HITS were identified in 92%. Eleven patients had a total count of <15 HITS, but in the remaining two the count was unexpectedly high (63 and 265 HITS). Echocardiography in these two patients indicated the presence of an atrial right-to-left shunt. Uneventful preoperative and postoperative visual function was found in 11 patients. One patient had preoperative blindness and in another ophthalmologic complications developed not related to microembolization. Scoliosis surgery is frequently associated with low counts of cerebral microemboli. It appears that such low embolic counts have no effects on postoperative visual function as determined clinically. Some patients may show high rates of microemboli, which may be related to the presence of right-to-left cardiac shunts. The impact of these signals on brain function remains to be investigated. PMID- 11433171 TI - Mouse model for thoracic congenital scoliosis. AB - SUMMARY: This study sought to produce a dose-response curve for acute and chronic maternal carbon monoxide (CO) exposure versus vertebral anomalies in mouse offspring and to determine the critical day of exposure. In Part I, pregnant CD-1 mice were exposed to an acute dose of CO at 9 days of gestation. A positive dose response relationship of acute maternal CO exposure and vertebral anomalies in the offspring was produced. In Part II, pregnant females were exposed to chronic CO for the first 11 days of gestation. Chronic exposure to CO did not produce significant vertebral anomalies. In Part III, pregnant females were exposed to an acute dose of 600 ppm of CO at gestation day 8, 9, or 10. Day 9 in this mouse breed is the critical day for maternal exposure to CO. The detected anomalies were predominately in the thoracic spine. PMID- 11433172 TI - Relationship between the patellar height and the disorder of the knee extensor mechanism in immature athletes. AB - SUMMARY: The purposes of this study were to evaluate the outcome of longitudinal research on the patellar height of immature athletes, and to clarify the relationship with disorders of the knee extensor mechanism. The patellar height of 40 young male soccer players (80 knees) was observed with radiographs of the bilateral knee joint for 2 years. The patellar height was measured using the epiphyseal line method. No subject showed patella alta before the occurrence of the disorder. Of the 12 knees in which the disorder occurred during the observation period, 10 knees (83%) had higher patella after the onset of symptoms than before. The patellar height in the painful stage was considered significant compared with the normal group. Although the patellar position in the predisorder stage was relatively high, this state was considered within the normal scope of physiologic variation, such as the growth spurt. PMID- 11433173 TI - Early radiographic differentiation of infantile tibia vara from physiologic bowing using the femoral-tibial ratio. AB - SUMMARY: The authors hypothesized that the ratio of the femoral to tibial metaphyseal-diaphyseal angles (femoral-tibial ratio [FTR]) more accurately differentiates physiologic bowing from infantile tibial vara than the tibial metaphyseal-diaphyseal angle (TMDA). The purpose of this study was threefold: to determine the false-negative and false-positive error rate of the FTR and TMDA; to determine to the effect of rotation on the FTR and TMDA; and to determine the reliability of the FTR and TMDA measurements. An FTR < 1 resulted in a false negative error rate of 10% and a false-positive error rate of 7%, whereas a TMDA > 13 degrees resulted in a false-negative error rate of 23% and a false-positive error rate of 10%. The difference between internal and external rotation was not significant for the FTR, whereas it was for the TMDA. The FTR was found to have good interobserver and intraobserver reliability (0.78 and 0.98, respectively). PMID- 11433174 TI - Growth in pediatric orthopaedics. PMID- 11433175 TI - Treatment of late-onset tibia vara using afghan percutaneous osteotomy and orthofix external fixation. PMID- 11433177 TI - Imagining the brain. PMID- 11433178 TI - Regulation of preglomerular microvascular 20-hydroxyeicosatetraenoic acid levels by salt depletion. AB - BACKGROUND: 20-hydroxyeicosatetraenoic acid (20-HETE) is the preeminent renal eicosanoid. The protean properties of 20-HETE - vasoactivity, mitogenicity and modulation of transport in key nephron segments - serve as the basis for the essential roles of 20-HETE in the regulation of the renal circulation and electrolyte excretion and as a second messenger for endothelin-1 (ET-1) and a mediator of selective renal effects of angiotensin II (AII). Renal autoregulation and tubular glomerular feedback are mediated by 20-HETE through constriction of preglomerular microvessels, particularly afferent arterioles. METHODS AND RESULTS: We had reported that rat preglomerular microvessels (PGMV; afferent interlobular-arcuate/interlobular) in response to angiotensin II (AII) generate primarily 20-HETE and lesser quantities of 19-HETE. We have now addressed a possible link between the renin-angiotensin-system (RAS) and induction of cyclooxygenase (COX-2). As Na+ deprivation induces COX-2 expression/activity in the renal cortex and AII stimulates release of 20-HETE from PGMV, we used a stimulus, low dietary salt, to activate the RAS and COX-2 and thereby explore potential interactions involving 20-HETE and COX-2. CONCLUSIONS: The capacity of COX to metabolize 20-HETE to prostaglandin analogs e.g., 20-OH PGF2a and 20-OH PGE2, may be critical to modifying the renal vascular and tubular actions of the eicosanoids. PMID- 11433179 TI - NCX4016 (NO-aspirin) inhibits thromboxane biosynthesis and tissue factor expression and activity in human monocytes. AB - BACKGROUND: NCX4016 (2 acetoxy-benzoate 2-(2-nitroxymethyl)-phenyl ester, NicOx S.A., France) is an antithrombotic agent chemically related to acetylsalicylic acid (ASA). We hypothesised that NCX4016, being able to release nitric oxide (NO) and to inhibit cyclo-oxygenase, might inhibit the prothrombotic function in human monocytes. MATERIAL AND METHODS: The effects of NCX4016 and ASA on the release of thromboxane (TX) B2 and tissue factor expression and activity were compared using adherent human monocytes. The tested drugs were added before stimulation with 10 Kg/ml LPS and incubation lasted 6 hours. TXB2 concentration was measured by RIA in the supernatant of cultured cells. Immunoreactive tissue factor (TF) concentration was determined by enzyme-linked immunoassay and TF activity was assayed by measuring the peptidyl activity of the tissue factor/ factor VII complex. RESULTS: Both ASA and NCX4016 10-300 Kmol/L dose-dependently reduced TXB2 release. NCX4016 activity was comparable to that of equimolar ASA. Part of the activity of NCX4016 up to 100 Kmol/L was prevented by 10 Kml/L ODQ, inhibitor of cGMP generation. Immunoreactive TF was dose-dependently inhibited by 300 Kmol/L NCX4016, but not by ASA. Also tissue TF activity was reduced by 300 Kmol/L NCX4016, but not by ASA. CONCLUSIONS: The present results indicate that NCX4016 not only has anti-platelet effects but also inhibits prothrombotic activities in human monocytes, partly via NO-dependent mechanisms. NCX4016 may prove effective in the clinical setting of athero-thrombosis. PMID- 11433180 TI - Role of epoxyeicosatrienoic acids as endothelium-derived hyperpolarizing factor in bovine coronary arteries. AB - BACKGROUND: Acetylcholine and bradykinin cause endothelium-dependent hyperpolarization and relaxation of vascular smooth muscle. These responses are mediated in part by an endothelium-derived hyperpolarizing factor (EDHF). Epoxyeicosatrienoic acids (EETs) are cytochrome P450 metabolites of arachidonic acid and appear to function as EDHFs in the coronary artery. This conclusion is based on the findings that EETs are synthesized by the vascular endothelium, open calcium-activated potassium (KCa) channels, hyperpolarize and relax vascular smooth muscle. METHODS: Bovine coronary arteries were precontracted with U46619 and isometric tension was measured to agonists. Whole cell K currents were measured in coronary smooth muscle cells by the patch clamp method. RESULTS: Bradykinin caused a concentration-related relaxation (ED50=10(-10) M) which was not affected by inhibitors of cytochrome P450. Inhibition of NO synthase and cyclooxygenase reduced the response to bradykinin (ED(50)=2 x 10(-9) M) and these responses were then completely blocked by cytochrome P450 inhibitors. 11, 12-EET relaxed the precontracted bovine coronary artery with an ED50 of 3 x 10(-8) M in the presence and absence of the endothelium. 14, 15-EET caused an increase in outward potassium current that was blocked by the KCa channel inhibitor iberiotoxin. A series of 14, 15-EET analogs were tested for agonist activity. CONCLUSIONS: These data indicate that cytochrome P450 metabolites contribute to bradykinin-induced, endothelium-dependent relaxations but only following inhibition of NO synthase and cyclooxygenase. EETs relax coronary arteries in an endothelium-independent manner. Carbon-1 carboxyl, 8, 9 and 11, 12-double bonds and 14, 15-epoxy groups are required for full agonist activity. These studies provide further support for the hypothesis that EETs represent EDHFs in the coronary artery and mediate the relaxation responses to bradykinin. PMID- 11433181 TI - Endothelial cell dysfunction in a model of oxidative stress. AB - BACKGROUND: We have investigated the role of L-arginine in hyperhomocysteinemia (HHCY). L-arginine is the substrate required for NO production by endothelial NOS (eNOS). When L-arginine is limited, NOS acts principally upon O2 to form superoxide (O2.-). Because HHCY causes formation of reactive oxygen species and reduced endothelial-dependent vasodilation, we hypothesized that HHCY decreases NO formation by limiting the cellular supply of L-arginine. MATERIAL AND METHODS: Studies with cultured bovine aortic endothelial cells (ECs) determined effects of HCY on transport of [3H] L-arginine. Effects on L-arginine transporter protein CAT-1 and eNOS protein were assessed by immunoblotting. Peroxynitrite formation was evaluated by an immunoassay for nitrotyrosine levels. eNOS activity in forming NO was determined by assay for 3H-L-arginine to 3H-citrulline conversion. RESULTS: HCY had a depressive effect on arginine transport in ECs. HCY treatment for a 24 hr period decreased arginine uptake by 27%. HCY treatment for 24 hr significantly reduced cellular levels of the CAT-1 arginine transporter protein ( approximately 30%) and increased nitrotyrosine formation, whereas levels of eNOS protein and basal NOS activity were not altered. Nevertheless, total NO production as indicated by citrulline conversion was significantly decreased. Treatment with the antioxidant N-acetylcysteine reversed the HCY effect on arginine transport, suggesting that transporter oxidation may contribute to the endothelial dysfunction. CONCLUSIONS: The association of HCY-induced decreases in NO formation with decreases in function and expression of the arginine transporter in the absence of alterations in eNOS expression or activity suggests a primary role for arginine transport alterations in HHCY. The action of HCY to reduce arginine uptake may accentuate endothelial dysfunction due to generation of O2.- and peroxynitrite formation, which may cause further oxidative injury. PMID- 11433182 TI - Comparison of nitric oxide-releasing NSAID and vitamin C with classic NSAID in healing of chronic gastric ulcers; involvement of reactive oxygen species. AB - BACKGROUND: Nonsteroidal anti-inflammatory drugs such as aspirin (ASA) are known to induce gastric mucosal damage including bleeding, ulceration and perforation in humans and experimental animals. These adverse effects of ASA were originally attributed to the inhibition of cyclooxygenase and the deficiency of endogenous prostaglandins induced by this drug but the role of reactive oxygen species (ROS), lipid peroxidation and antioxidizing mechanism in the pathogenesis of ASA damage has been little studied. New class of nitric oxide (NO)-releasing nonsteroidal anti-inflammatory drugs was shown to inhibit cyclooxygenase and prostaglandin generation without causing mucosal damage but it remains unknown whether these agents affect the healing process of chronic gastric ulcers. MATERIAL AND METHODS: In this study the effect of NO-releasing aspirin (NO-ASA) and was compared with that of native aspirin applied with or without vitamin C on the healing of acetic acid ulcers. The area of gastric ulcer was determined by planimetry, the gastric blood flow (GBF) at ulcer margin was measured by H2 gas clearance method and mucosal release of ROS was quantified by measuring the chemiluminescence before and after the treatment with ASA or NO-ASA alone and ASA combined with vitamin C. The plasma antiinflammatory cytokine such as IL-1b and oxygen radical-mediated lipid peroxidation was measured in the ulcerated gastric mucosa of ASA and NO-ASA-treated animals. RESULTS: ASA delayed significantly ulcer healing and this effect was accompanied by a marked increase in the chemiluminescence, lipid peroxidation and the fall in the GBF at ulcer margin. Vitamin C attenuated significantly both the ASA-induced gastric damage and accompanying fall in the GBF at ulcer margin and the rise in the chemiluminescence and reversed the ASA-induced lipid peroxidation. In contrast, NO-ASA failed to affect healing of gastric ulcers and failed to produce the rise in the plasma IL-1b levels and the increase of lipid peroxidation as compared to those recorded in ASA-treated animals. CONCLUSIONS: 1) ROS-induced enhancement in lipid peroxidation plays an important role in the mechanism of gastric damage induced by ASA, 2) vitamin C attenuates the deleterious effect of ASA on ulcer healing due to its antioxidizing activity by mechanism involving preservation of gastric microcirculation and attenuation of lipid peroxidation and cytokine release and 3) coupling of NO to aspirin fails to delay the ulcer healing suggesting that NO might compensate for prostaglandin deficiency induced by NSAID. PMID- 11433183 TI - Studies on the antithrombotic action of AT1 receptor antagonists. AB - BACKGROUND: In our previous experiments we showed that the prototype member of the AT1 receptor antagonists (AT1-As) family, losartan, prevented the development of arterial and venous thrombosis in rats. Recent studies have demonstrated that apart from blocking AT1 receptor, losartan is also a competitive antagonist to thromboxane A2/prostaglandin H2 receptor (TP receptor). Thus, we decided to assess if this feature could contribute to the antithrombotic action of losartan. MATERIAL AND METHODS: We compared the influence losartan, its active metabolite EXP3174 and valsartan on rat platelet adhesion to fibrillar collagen and platelet aggregation in response to thromboxane A2 analogue, U46619. We also assessed the efficacy of these drugs in platelet-dependent pulmonary thrombosis in mice as well as preventive and therapeutic models of venous thrombosis in rats. RESULTS: All the three compounds, given in a single dose, inhibited rat platelet adhesion to fibrillar collagen and platelet aggregation induced with U46619 in vitro and ex vivo, with the action of losartan being much more pronounced than that of EXP3174 or valsartan. Losartan also more effectively protected mice from death in response to the intravenous injection of collagen / epinephrine and it was the only compound which reduced mice mortality after the intravenous injection of U46619. In contrast, all the three AT1 receptor antagonists exerted a similar thrombolytic action and comparably decreased the thrombus weight in the therapeutic and preventive model of venous thrombosis, although in the latter case a high dose of losartan was slightly more effective than a corresponding dose of EXP3174 and valsartan. CONCLUSIONS: Since losartan is endowed with a relatively low affinity towards the AT1 receptor, we conclude that its superiority over EXP 3174 and valsartan in inhibiting thrombocyte function and platelet-dependent thrombosis could result from its stronger action on the TP receptor. This feature seems to be less important in the thrombolytic effect of AT1-As and in the inhibition of the venous thrombosis development, in which platelets play only a minor role. PMID- 11433184 TI - Detection of protein oxidation in endothelial cells by fluorescently labelled tyramine. AB - BACKGROUND: Endothelial cell injury mediated by activated polymorphonuclear leucocytes (PMN) occurs during inflammation or reperfusion after brain ischemia. Protein oxidation caused by activated PMN may lead to functional disturbances, degeneration and death of the endothelial cells. The aim of this study was to detect protein oxidation in endothelial cells induced by activated neutrophils by using a novel fluorescent probe. MATERIAL AND METHODS: Protein oxidation of Human Umbilical Vein Endothelial Cells (HUVEC) in culture was investigated by a 15-min incubation with human neutrophils activated by phorbol myristate acetate (PMA) in the presence of tyramine coupled to the succinimidyl ester of (fluorescein -5 (and-6)-carboxamido) hexanoic acid. Dityrosine bond formation as reflected by the linkage of the fluorescent tyramine to proteins was determined by Western blotting. RESULTS: The oxidative burst generated by activated neutrophils induced dityrosine formation in the extracellular proteins (ECP) of HUVEC. Similar results were obtained, when horseradish peroxidase (HRP) was used for the induction of oxidative stress. However, when hydrogen peroxide (0.1 mM) was used, dityrosine formation was not detected. CONCLUSIONS: Fluorescently labelled tyramine is a powerful tool for the detection of ECP oxidation in endothelial cells. As long as the oxidation by the activated neutrophils is limited to ECP, the endothelial cells may be protected by antioxidants. PMID- 11433185 TI - Gastric preconditioning induced by short ischemia: the role of prostaglandins, nitric oxide and adenosine. AB - BACKGROUND: Various organs including heart, kidneys, liver or brain respond to brief exposures to ischemia with an increased resistance to severe ischemia and this phenomenon is called 'preconditioning'. No study so for has been undertaken to check whether such short, repeated gastric ischemic episodes protect gastric mucosa against the damage caused by subsequent prolonged ischemia-reperfusion or necrotizing substances. MATERIAL AND METHODS: In this study, cyclooxygenase (COX) 1, COX-2, nitric oxide (NO) and adenosine receptors inhibitors were used to determine the possible involvement of endogenous prostaglandin, NO and adenosine in the mechanism of gastric preconditioning. This ischemic preconditioning was induced by short episodes of occlusion of celiac artery from 1 to 5 times, for 5 min each applied 30 min before prolonged (30 min) ischemia followed by 3 h of reperfusion (I/R) or 30 min before topical application of strong mucosal irritants such as 100% ethanol, 25% NaCl or 80 mM taurocholate. RESULTS: Exposure to regular I/R produced numerous gastric lesions and significant fall in the gastric blood flow and PGE2 generation. Short (5 min) ischemic episodes even induced several times (1-5 times) by itself failed to cause any gastric lesions but significantly attenuated those produced by I/R and this protective effect reached maximum with two 5 min ischemic episodes and this preconditioning was considered as standard. The protective effects of standard ischemic preconditioning against gastric lesions induced by I/R was accompanied by a reversal of the fall in the gastric blood flow and PGE2 generation and resembled those induced by classic gastric mild irritants such as 20% ethanol, 5% NaCl and 5 mM taurocholate. These protective and hyperemic effects of standard preconditioning, lasted up to 6-8 h, and were significantly attenuated by pretreatment with specific COX-1 and COX-2 inhibitors such as Vioxx (5 mg/kg i.g.) and resveratrol (10 mg/kg i.g.) that failed to affect PGE2 generation in intact gastric mucosa but attenuated significantly that in preconditioned gastric mucosa. Non-specific COX-inhibitor indomethacin (5 mg/kg i.p.), that suppressed the PGE2 generation by approximately 90% and non-specific NO synthase inhibitor L NNA (20 mg/kg i. p.), that significantly suppressed NO production, significantly inhibited the protection and the rise in GBF induced by standard preconditioning and these effects were restored by addition of 16,16 dm PGE2 (1 Kg/kg i.g.) or L arginine (200 mg/kg i.g.), a substrate for NO-synthase, to indomethacin or L NAME, respectively. Pretreatment with adenosine (10 mg/kg i.g.) also reduced the lesions induced by I/R and increased the gastric blood flow with the extent similar to that observed with standard ischemic preconditioning, while an antagonist of adenosine receptors, 8-phenyl theophylline (SPT, 10 mg/kg i.g.) attenuated significantly the gastroprotection afforded by the preconditioning. Gene expression of COX-1 but not COX-2 was detected by RT-PCR in intact gastric mucosa and in that exposed to I/R with or without ischemic preconditioning, whereas COX-2 was overexpressed only in preconditioned mucosa. CONCLUSIONS: 1) gastric ischemic preconditioning represents one of the most powerful protective intervention against the mucosal damage induced by severe I/R as well as by topical mucosal irritants in the stomach; 2) this protection, involving several mediators such as PG derived from COX-1 and COX-2, NO originating from NO synthase and adenosine, appear to play a key mechanism of gastric ischemic preconditioning. PMID- 11433186 TI - Statin-stimulated nitric oxide release from endothelium. AB - BACKGROUND: There is increasing evidence that loss of endothelium-derived NO is a major factor in cardiovascular complication events, and that NO might exert antiatherosclerotic actions. The beneficial effects of HMG CoA reductase inhibitors (statins) therapy in atherosclerosis outweigh those expected from simply lowering low-density lipoprotein (LDL) cholesterol, and may be related to the direct action in the endothelium. Based on these concepts, in the studies described here, the effect of new statin derivatives on nitric oxide (NO) and superoxide (O2-) release in bovine endothelial cells was tested. MATERIAL AND METHODS: Highly sensitive electrochemical NO and O2--microsensors were placed near the surface of endothelial cells, and the concurrent kinetics of NO and O2-- release were measured in situ. RESULTS: All tested statins stimulated NO release. The peak concentration of NO after stimulation with 1 Kmol/l Lovastatin, 1 Kmol/l Atorvastatin, 1 Kmol/l Pravastatin, or 1 Kmol/l Simvastatin was about 77%, 73%, 72%, and 44% lower, respectively, as compared with the NO peak concentration after stimulation with 1 Kmol/l calcium ionophore A23187 (receptor-independent agonist). The tested statins stimulated NO release in a modest way, which resulted in diminishing O2- generation during activation of nitric oxide synthase. Moreover, the kinetics of O2- release after administration of the statins suggested that these compounds may also scavenge O2-. The NO/O2- peak concentration ratio after the NOS agonists administration was as follows: 7.51 for CaI, 6.56 for Lovastatin, 6.00 for Atorvastatin, 4.17 for Pravastatin and 6.25 for Simvastatin. CONCLUSIONS: The tested statins, i.e. Lovastatin, Atorvastatin, Pravastatin and Simvastatin demonstrate variable potency to enhance the NO/O2- concentration ratio after stimulation of NOS, resulting in an increase of NO bioavailability in endothelial cells. PMID- 11433187 TI - Studies on potential involvement of protein kinase C in glomerular insensitivity to atrial natriuretic factor on low sodium intake. AB - BACKGROUND: Atrial natriuretic factor (ANF)-induced increase in glomerular filtration rate (GFR) is inhibited on low sodium intake. It has been shown that activation of renin-angiotensin system on low sodium intake antagonizes the biological effect of ANF by interfering in the intracellular metabolism of cGMP. We have previously indicated that the renin-angiotensin system increases activity of Ca2+/calmodulin dependent-cyclic GMP phosphodiesterase (cGMP-PDE) in glomeruli and thereby inhibits the ANF-induced increase in GFR in low sodium-treated rats. The aim of the present study was to investigate whether low sodium intake might change glomerular cGMP metabolism by the alternative branch of the signal transduction pathway, namely protein kinase-C (PKC) activation. MATERIAL AND METHODS: cGMP formation and PKC activity were examined in isolated glomeruli from the rats maintained for five days on a normal or a low sodium diet. Renal hemodynamic parameters in clearance experiments during infusion of ANF (0.5 Kg/min/kg body weight) in both groups of rats were also evaluated. RESULTS: Low sodium intake inhibited ANF-dependent increase in GFR and nephrogenous cGMP excretion, whereas urinary sodium excretion did not differ appreciably in rats on either diet. The basal and ANF-stimulated cGMP formation in isolated glomeruli was significantly inhibited in low sodium-treated rats as compared to normal sodium-treated rats. The inhibitory effect of low sodium intake on basal and ANF stimulated glomerular cGMP formation was completely prevented by a selective cGMP PDE inhibitor, zaprinast, but not affected by PKC activator, PMA, or PKC inhibitor, H-7. The activity of PKC in glomeruli neither in membrane fraction nor in cytosol fraction did not differ significantly between normal and low sodium treated rats. CONCLUSIONS: These results demonstrate that the blunted glomerular response to ANF in rats on low sodium intake is due to decrease ability of cGMP formation in glomeruli by increasing activity of cGMP-PDE without altering activity of PKC. PMID- 11433188 TI - The role of P2Y-receptors in the regulation of glomerular volume. AB - BACKGROUND: Extracellular ATP signaling affects the cells of renal glomeruli via activation of P2-purinoceptors, denoted as P2X and P2Y. Through either of these purinoceptors, ATP is able to stimulate an increase in intracellular [Ca2+]. P2Y receptors are expressed on mesangial and endothelial cells, thus may participate in contraction and relaxation of glomeruli, respectively. Moreover, P2Y-receptors possess activity of ecto-ATPase which may lead to dephosphorylation of ATP and generation of adenosine. The aim of the present study was to investigate the involvement of P2Y-receptors in responses of renal glomeruli to extracellular ATP. MATERIAL AND METHODS: Renal glomeruli were isolated from rats by sieving technique. [3H]-inulin was used to measure the intracapillary volume of isolated glomeruli. Changes of intracapillary volume reflect contraction and relaxation of the glomeruli. ATP and adenosine concentration in the incubation mixture were measured using luminometric methods. RESULTS: Extracellular ATP (1 microM) induced relaxation of Ang II-precontracted glomeruli in time-dependent manner. The glomeruli relaxed completely at 2nd minute of incubation. The relaxation was considerably diminished at 5th minute of incubation as compared to 2nd minute. Relaxing effect was completely prevented by an antagonist of P2Y-receptors i.e. reactive blue 2. The decrease in ATP concentration with time was accompanied by a rise in adenosine concentration which led to contraction of glomeruli. Non metabolised analogue of ATP, an agonist of P2Y-receptors i.e. 2-methylthio-ATP (1 microM) induced complete relaxation at 2nd minute of incubation but there was no effect at 5th minute of incubation. CONCLUSIONS: The extracellular ATP through activation of P2Y-receptors may regulate the volume of renal glomeruli, which in turn influences on the glomerular filtration rate, through at least two mechanisms: one is ATP-dependent glomerular relaxation in the initiate phase and the other is glomerular contraction caused by either ATP itself or adenosine formed from ATP hydrolysis in maintenance phase. PMID- 11433189 TI - Suppression of basal NO- and prostacyclin-dependent tone in cerebral microcirculation during chronic hyponatremia in female rats. AB - BACKGROUND: Our previous studies in the rat model of chronic hyponatremia have demonstrated the decrease of cerebral blood flow along with the attenuation of cerebral metabolic rate for oxygen exclusively in females and when hyponatremia was induced with vasopressin. Present study was designed to verify the hypothesis that the decrease of cerebral blood flow during chronic vasopressin-induced hyponatremia in female rats is due to the attenuation of basal vasodilator tone produced in cerebral microcirculation under physiological conditions by shear stress-dependent release of nitric oxide and prostacyclin. MATERIAL AND METHODS: The experiments were performed on 47 anesthetized and mechanically ventilated adult, female Wistar rats. Chronic hyponatremia (3.5 days) was induced in these rats by the twice-daily subcutaneous administration of vasopressin (AVP) in conjunction with 140 mmol/L glucose/water solution. Cerebrocortical microflow (LDF) was monitored using laser-Doppler probe. LDF responses to the administration of either NG-nitro-L-arginine methyl esther (L-NAME, 30 mg/kg i.v.), indomethacin (INDO, 6 mg/kg i.v.), acetylcholine (ACh, 10 Kl/20 Kl/min ica) or 5% CO2 were tested in normonatremic and hyponatremic rats. RESULTS: The response of LDF to L-NAME, INDO and ACh were abolished in rats with hyponatremia. In contrast, CO2 reactivity was well preserved. CONCLUSIONS: Our results demonstrate that attenuation of cerebral blood flow during chronic AVP-induced hyponatremia in female rats is, at least in part, due to the withdrawal of basal vasodilator tone produced in cerebral circulation under physiological conditions by nitric oxide and prostacyclin. PMID- 11433190 TI - Matrix metalloproteinase-2 in platelet adhesion to fibrinogen: interactions with nitric oxide. AB - BACKGROUND: Matrix metalloproteinase-2 (MMP-2) has been shown to activate a non thromboxane, non-ADP pathway of platelet aggregation. In contrast, nitric oxide (NO) is known to inhibit platelet adhesion and aggregation. Therefore, we have studied the release of MMP-2 during platelet adhesion to fibrinogen, the effects of phenanthrolione, an MMP-2 inhibitor, on adhesion and the interactions of inhibitor with a NO donor, S-nitroso-N-acetyl-D, L-penicillamine (SNAP). MATERIAL AND METHODS: Human platelets were isolated from blood of healthy volunteers and platelet adhesion to fibrinogen-coated plates was studied by measuring thrombin stimulated release of platelet a-granule constituent, platelet factor 4. In addition, the mode of action of phenanthroline and NO on platelets was investigated by assaying the levels of intraplatelet cyclic GMP. RESULTS: Thrombin-stimulated platelet adhesion to fibrinogen was associated with increased release of MMP-2 from platelets. Phenanthroline (0.1-100 KM) reduced platelet adhesion to fibrinogen. The adhesion was also inhibited by SNAP (0.1-100 KM), an effect abolished by 1H-[1,2,4] oxadiazolol [4,3,-a] quinoxalin-1-one (ODQ), a selective inhibitor of the soluble guanylate cyclase. Co-administration of phenanthroline and SNAP resulted in a synergistic inhibition of platelet adhesion, an effect that was not associated with enhanced cyclic GMP generation by platelets. Furthermore, ODQ did not reverse the synergistic effect of these compounds on adhesion. CONCLUSIONS: 1. MMP-2 promotes platelet adhesion to fibrinogen. 2. Phenanthroline and NO synergize to inhibit platelet adhesion to fibrinogen acting through a cyclic GMP-independent mechanism(s). PMID- 11433191 TI - In vitro response of phagocytic cells to immunomodulating agents. AB - BACKGROUND: Phagocytes (polymorphonuclear cells and monocyte-macrophages) are the first line of defence of the host against infectious microorganisms and other foreign antigens. Agents which participate in activation of phagocytic cells possess a potential immunomodulating action. Thus, search for convenient in vitro test-systems and study of mechanisms of action of these agents are of practical interest. MATERIAL AND METHODS: Human blood polymorphonuclear (PMN) cells and murine macrophages (line J774.2) were used as cellular test-systems for study of phagocytosis-stimulating action of immunomodulating agents. Indexes of phagocytic activity were estimated by the phagocyte ingestion of yeast cells. NO-synthase activity, nitrite production, and nitroblue tetrazolium test were determined after phagocyte stimulation. RESULTS: It was revealed that indexes of phagocytic activity can be used as quantitative indicators for measurement immunomodulating activity. Zymosan A-induced phagocytosis in almost 100% PMN cells and macrophages and thus can be used as a positive control. Wheat germ agglutinin (WGA, 0.5-1.0 microg/ml) stimulated phagocytosis in PMN cells 1.8 times after 2-3 h incubation, although in higher concentrations (5-10 microg/ml) it strongly inhibited phagocytosis. TGF-b1 (10 ng/ml) suppressed phagocytosis in WGA-stimulated PMN cells. Mistletoe agglutinin-1 stimulated phagocytosis in PMN cells, although its effect in macrophages was weak, while concanavalin A stimulation of phagocytosis in macrophages was well expressed. Vasodilating peptide bradykinin increased phagocytosis 2.5 times in macrophages. We did not reveal changes in NO-synthase activity and nitrite production in macrophages and PMN cells activated by different immunomodulatig agents. Only lipopolysacharide stimulated such activity in macrophages. CONCLUSIONS: Cultured macrophages and PMN cells can provide reproducible quantitative results in screening phagocytic activity of different immunomodulating agents. Both positively and negatively acting immunomodulators might be studied using these test cells. PMID- 11433192 TI - Cellular heterogeneity of rat vascular endothelium as detected by HPA and GS I lectin-gold probes. AB - BACKGROUND: In order to extend previous observations on the heterogeneity in labeling of rat vascular endothelium by the lectins GS I and LEA [1,2], we have conducted a survey of several organs with a lectin panel with greater variety of carbohydrate specificity. MATERIAL AND METHODS: Submandibular gland, duodenum, distal colon, liver, kidney, heart, skeletal muscle, adrenal gland, ovary, cerebral and cerebellar cortex of a rat have been examined by light and electron microscopy using lectin-gold probes on Lowicryl K4M embedded tissue. RESULTS: Five lectins (LCA, con A, LEA, RCA, WGA) labeled vascular endothelial cells in the connective tissue of all the organs studied, while PNA, EEA, TPA, LABA, UEA I expressed no detectable affinity towards endothelial cells. HPA and GS I reacted with most endothelial cells, except those in the kidney glomeruli, liver sinusoids and zona fasciculata of adrenal gland. In cerebral and cerebellar cortex GS I reacted witn pericytes of large vessels, but not with endothelial cells of the capillary bed. SDS-PAGE extracts of heart, skeletal muscle and cerebral cortex reveal that differences in GS I labeling depend, at least in part, on 40 and 200 kD glycoproteins, present in heart and skeletal muscle, but not cerebral cortex. CONCLUSIONS: Data indicate, that GS I, widely used for selective histochemical labeling of rat endothelial cells is not uniform endothelial marker in all organs. More precise investigation of these lectin reactive determinants in rat vascular endothelium, including developmental changes, isolation and enzyme-FACE sequencing of their carbohydrate moieties, generation of antibodies and their possible organ specific binding affinities could give new insight into the physiological role of GS I and HPA binding glycoproteins in rat endothelium. PMID- 11433193 TI - Receptor-mediated activation of the heterotrimeric G-protein Gs results in ATP release from erythrocytes. AB - BACKGROUND: Adenosine triphosphate (ATP), released from the erythrocyte in response to mechanical deformation, decreased oxygen tension or reduced pH, has been suggested to be an important determinant of vascular resistance in several vascular beds. Mechanical deformation-induced ATP release from rabbit and human erythrocytes was reported to require the activity of the cystic fibrosis transmembrane conductance regulator (CFTR), suggesting that a signal transduction pathway involving CFTR mediates ATP release from erythrocytes. Here we investigate the hypothesis that the heterotrimeric G-protein Gs is also involved in this signal transduction pathway. MATERIALS AND METHODS: The heterotrimeric G protein Gs was identified in rabbit and human erythrocyte membranes, using gel electrophoresis. The concentration of ATP released into a suspension of erythrocytes, incubated with iloprost or epinephrine, was measured using the luciferin/luciferase assay. RESULTS: The 45 kDa form of the heterotrimeric G protein Gs was identified in rabbit and human erythrocyte membranes. Incubation of rabbit erythrocytes with iloprost (n=18) or epinephrine (n=6) increased the ATP concentration by 106+/-16% and 156+/-54%, respectively. Epinephrine-induced changes in ATP concentrations were prevented by pretreatment with propranolol. CONCLUSIONS: The heterotrimeric G-protein Gs is present in erythrocyte membranes. Receptor-mediated activation of Gs results in ATP release. These results are consistent with the hypothesis that Gs is a component of a signal transduction pathway for ATP release from erythrocytes. PMID- 11433194 TI - Activity of cysteine protease inhibitors in human brain tumors. AB - BACKGROUND: Cysteine proteases (mainly cathepsins B and L) are thought to play an important role in the progress of cancer, including brain tumors. Together with other proteases, they hydrolyze the extracellular matrix and basement membrane proteins, thus enabling the tumor to grow and spread. Therefore cysteine protease inhibitors are regarded as protective factors, able to prevent tumor growth and dissemination. MATERIAL AND METHODS: In this study, the activity of cysteine protease inhibitors (CPIs) was investigated in material derived from patients with brain tumors (astrocytoma and meningioma). The activity of CPIs was measured as antipapain activity in tissue homogenates, cerebrospinal fluid, and serum, with N-benzoyl-DL-arginine-2-naphthylamide hydrochloride (BANA) as a substrate, according to Barret's method. RESULTS: Tumorous tissues showed higher activity of cysteine protease inhibitors than control tissues, but this difference proved to be statistically insignificant. The activity of CPIs was lower in cerebrospinal fluid and serum from patients with brain tumors. CONCLUSIONS: The activity of CPIs measured in brain tumor tissue cannot be taken as a marker of any type of tumor, whereas CPI activity in cerebrospinal fluid and serum may be considered a marker of meningioma. In meningioma patients the level of CPIs may be too low to prevent the host tissues from the growing tumor. PMID- 11433195 TI - Effect of novel platinum complexes on survival rate of B16 and Cl S91 melanoma cells and oxidation stress in vitro. AB - BACKGROUND: The aim of study was the assessment of two novel platinum complexes and their cytotoxic effect on murine melanoma cells. The evaluation of possible oxidation stress in neoplastic cells coexistent with the effect of these cytostatic agents. MATERIAL AND METHODS: Two lines of murine metastatic melanomas: B16 and Cl S91 were selected for the experiments. Two platinum compounds were tested: cis-dichloro(dimethylsulphoxide)(1-beta-D-ribofuranosyl 1,2,4-triazol-3-carboxyamide)platinum (II) called Pt-complex 1 and chloro(dimethylsulphoxide)(1-beta-D-ribofuranosyl-1,2,4-triazol-3 carboxyamide)platinum (II) called Pt-complex 2. Melanoma cells were incubated for 24 hours with the platinum complexes mentioned above and culture medium at the concentration of 0.01, 0.05 and 0.5 mg/ml. Survival rate of melanoma cells, superoxide dismutase (SOD) activity, the concentration of thiobarbituric acid reactive substances (TBARS) and the content of conjugated dienes were estimated. RESULTS: A number of statistically significant relationships between analysed parameters were observed. Negative correlation ratios were found between increasing levels of analysed platinum complexes and the survival rate of the cells of both melanoma lines as well as between the survival rate of Cl S91 melanoma cells and the level of lipid peroxidation products and SOD activity. On the other hand, there was a positive correlation between increasing platinum complexes and the level of lipid peroxidation products as well as between the survival rate of B16 melanoma cells and SOD activity and the content of CD. CONCLUSIONS: Oxygen free radicals may play an important role in the development of melanoma and cytostatic agents disturb the balance between generation and removal of reactive oxygen forms in melanoma cells. PMID- 11433196 TI - Depressive state and regional cerebral activity in cancer patients - a preliminary study. AB - BACKGROUND: The aim of this study was to investigate influences of depressive states, chemotherapy and existence of remaining tumors on the regional brain activity of cancer patients. MATERIAL AND METHODS: Positron emission tomography with 18F-fluorodeoxyglucose was performed on 21 patients with various types of cancer. Their brain images were compared to 10 age- and gender-matched control data using statistical parametric mapping (SPM). The patients were subgrouped into the with and without depression based on the scores on Zung's self-rating depression scale (SDS), with and without previous chemotherapy, and with and without existence of remaining tumors. RESULTS: Significant metabolic reduction was detected in the cingulate gyrus, prefrontal, dorsolateral prefrontal, temporoparietal cortices and basal ganglia in cancer patients. These findings were close to known lesions of major depression. Intra-group comparisons showed that these hypometabolic findings were associated with the depth of depressive state. Influences of chemotherapy and remaining tumors on the cerebral cortex seemed to be weaker than that of psychological factors. CONCLUSIONS: The present pilot study suggests that frontal hypoactivity commonly seen in cancer patients is likely to be associated with depression rather than chemotherapy or remaining tumors. A brain mapping technique might be useful in evaluating neuropsychiatric problems in cancer patients. PMID- 11433197 TI - Plasma leptin concentrations in patients with Graves' disease with or without ophthalmopathy. AB - BACKGROUND: Recent studies have demonstrated that human preadipocyte fibroblasts in orbital connective tissues from patients with Graves' ophthalmopathy differentiate into cells resembling adipocytes and acquire expression of leptin and functional thyroid stimulating hormone receptors. These finding imply that leptin may play a role in the pathogenesis of Graves' ophthalmopathy. However, little is known about plasma leptin concentration in patients with Graves' disease with or without ophthalmopathy. MATERIAL AND METHODS: To investigate this relationship; 28 patients with active Graves' ophthalmopathy (19 female and 9 male, mean age: 32.7+/-10.5 years, mean BMI: 24.8+/-3.7 kg/m2) and 10 patients without ophthalmopathy (6 female and 4 male, mean age: 24.6+/-5.6, mean BMI: 23.02+/-2.4 kg/m2) all with untreated Graves' disease were included in the study at first diagnosis in our endocrinology out-patient clinic. Sex-, BMI- and age matched twenty healthy subjects (13 female, 7 male, mean age: 31.9+/-10.0, mean BMI: 24.2+/-3.0 kg/m2) were selected as a control group. Plasma leptin levels were measured by a RIA method with a sensitivity of 0.5 ng/ml. RESULTS: Results showed any significant differences neither between patients and controls (7.97+/ 5.2 ng/ml vs. 7.83+/-3.7 ng/ml) nor between patients with or without Graves' ophthalmopathy (8.29+/-5.0 ng/ml vs. 7.06+/-5.8 ng/ml) (both P>0.05). Moreover, no correlation was found between plasma leptin levels and ophthalmopathy index score, or proptosis. CONCLUSIONS: Although effects of local leptin production in the orbit cannot be excluded, our data suggest that circulating plasma leptin does not have a significant direct influence on ophthalmopathy index score or pathophysiology of Graves' ophthalmopathy. PMID- 11433198 TI - Does alpha-1-proteinase inhibitor play a protective role in coronary atherosclerosis? AB - BACKGROUND: alpha-1-proteinase inhibitor (alpha-1-PI) might appear to be actively involved in atherogenesis as an important regulator protecting elastic tissue from damage by proteinases as well as lipid accumulation in arterial wall. MATERIALS AND METHODS: alpha-1-PI genetic variants were examined in 156 male coronary atherosclerosis patients (mean age 49+/-9 years). The frequency of alpha 1-PI phenotypes was determined in 1577 healthy individuals. Also, 108 long survivors (mean age 92.7+/-4.3 years) were investigated for alpha-1-PI phenotype. The serum level of alpha-1-PI was examined in 43 coronary atherosclerosis patients (mean of age 49.6+/-8.1 years) and in different age groups of healthy males. A correlations between seruma-1-PI concentration and apoB, and apoA-1 were calculated. Relationship between serum alpha-1-PI and apolipoprotein B/apolipoprotein A-1 (apoB/apoA-1) ratio was determined. Isoelectric focusing of alpha-1-PI phenotypes was performed on thick polyacrylamide gels with ampholytes pH 3.5-5.0, 4.5-6.5 and 4.2-5.0. A quantitative assessment of alpha-1-PI, apoB and apoA-1 was performed by ELISA method using antibodies against human alpha-1 PI, apoB and apoA-1. RESULTS: The phenotype PI ZZ of alpha-1-PI associated with a severe congenital deficiency of alpha-1-PI in patients and long-lived subjects was not found. In population this phenotype (0.06%) as well as PI SS phenotype (0.06%) was detected once. No significant difference was found in the frequency of PI*M, PI*S and PI*Z genes in the coronary atherosclerosis group and population. The PI*Z gene was significantly more frequent in patients with coronary atherosclerosis than in long-survivors (P<0.01). The correlation between blood serum (-1-PI concentration and age in the control group of males was significant (r=-0.83, P<0.01). The alpha-1-PI concentration was found significantly higher in coronary atherosclerosis patients compared with control (2.14+/-0.6 and 1.68+/-0.3 g/l respectively, P<0.05). The alpha-1-PI concentration was found to dependent on the apoB/apoA-1 ratio: it was higher in patients having apoB/apoA-1>1.0 than in patients with apoB/apoA-1<1.0 (2.4+/-0.6 and 2.04+/-0.5 g/l respectively, P<0.05). There was a significant correlation between alpha-1-PI concentration and apoB/apoA-1 ratio in blood serum of patients suffering from coronary atherosclerosis (r=0.25, P<0.05). CONCLUSIONS: Derangements in the homeostasis of proteinase-antiproteinase system could be involved in destruction of the arterial wall connective tissue. The local as well as systemic inactivation of alpha-1-PI in atherosclerosis process could be related with hyperlipidemia. Investigation of long-survivors support the suggestion that they are important antiatherogenic control group for evaluating the role of genetic determinants in atherogenesis. Congenital (alpha-1-PI deficiency, hyperlipidemia) and acquired (related to smoking, aging) imbalance of proteinase-antiproteinase system is considered to one of the atherogenic factors. Authors discuss the possible mechanisms of atherogenesis related with imbalance of proteinase-antiproteinase system. PMID- 11433199 TI - Is the spinal cord lesion rare in diabetes mellitus? Somatosensory evoked potentials and central conduction time in diabetes mellitus. AB - BACKGROUND: Diabetic neuropathy and autonomic nervous system neuropathy are recognized as the most common clinical pictures of nervous system disorders caused by DM, while the damage to the brain and the spinal cord is considered to be rare. The aim of this work is to indicate the importance of somatosensory evoked potentials (SEP) for the early diagnosis of nerve system damage related to diabetes mellitus. MATERIAL AND METHODS: We studied 20 patients (aged 35-50 years), with type 2 diabetes mellitus lasting for 5-10 years. The control group population comprised of 30 healthy individuals of the same median age. The spinal and cortical somatosensory evoked potentials (SEP) after stimulation of median and fibular nerves were examined in both groups of patients. We measured the latencies and amplitudes of individual wave deflections, peripheral and central conduction time (PCT and CCT) of spinal and cortical SEP. RESULTS: The examination proved and confirmed the elongation not only of peripheral conduction time but also of the central conduction time - especially in spinal cord structures. The spinal cord changes connected with decrease of myelinated fibers which are able of conduction from periphery have to be accepted. The comparison of CCT1 and CCT2 conduction times showed that in group of diabetic patients the spinal cord structures are responsible for prolongation CCT. CONCLUSIONS: We confirm that the use of somatosensory evoked potentials examination and conduction times measurement has the significance in the confirmation of inapparent lesion of the spinal cord in diabetics. PMID- 11433200 TI - Disturbances of mental image processing in post-stroke patients with left and right hemisphere damage. AB - BACKGROUND: The purpose of this article is to point out significant differences in how mental images are processed by post-stroke patients with left and right hemisphere damage. The issues involved are of theoretical importance because of the light shed on the modularity of cerebral functions, especially the imagination, and of clinical importance due to the better understanding of the underlying pathomechanism. MATERIAL AND METHODS: The research involved 82 right handed patients with a lesion in the left hemisphere (Group L), 82 right-handed patients with a lesion in the right hemisphere (Group R), and, as a control group, 82 patients with musculo-skeletal disorders not affecting the central nervous system (Group C), matched by age and sex. Image processing of complex notions was examined by using selected items from the Simple Neurolinguistic Test. RESULTS: In the control group, the majority of the patients responded to most of the prompts with polymodal associations of various types. In Group L, responses were dominated by isolated elements of the complex situation, while in Group R the associations were mostly verbal (lexical) and highly restricted in scope. CONCLUSIONS: The results indicate that the loss of LH functions interferes with the ability to assemble pieces of polymodal image information into sensible strings, while the loss of RH functions leaves strings to which little information is attached. PMID- 11433201 TI - Effect of scalp infiltration with lidocaine on the circulatory response to craniotomy. AB - BACKGROUND: The purpose of the present study was to evaluate the influence of infiltration anesthesia of the projected line of incision in the skin of head on the hemodynamic response of the circulatory system, and the essential dose of opioids in 100 patients who underwent craniotomy. MATERIAL AND METHODS: The patients were divided into 4 groups: IA - brain tumors, no infiltration anesthesia; IB - cerebral aneurysms, no infiltration anesthesia; IIA - brain tumors + infiltration anesthesia; IIB - cerebral aneurysms + infiltration anesthesia. In the patients from groups IIA and IIB, the projected line of skin incision was injected with a 1% lidocaine solution (9.94+/-1.95 ml) five minutes before commencing surgery. RESULTS: No statistically significant differences were found between the various groups in regards to their weight, the time between infusion of the first dose of fentanyl and skin incision, the mean heart rates at time points T1 (prior to induction of anesthesia), T2 (prior to skin incision) or T3 (after skin incision), or mean arterial pressure values (MAP) at time points T1 or T2. A significant increase in MAP values (P<0.05) caused by skin incision was recorded in groups IA (from 95.2+/-9.85 to 119+/-10.6 mm Hg) and IB (from 88.4+/-11.5 to 100.3+/-11.4 mm Hg). In group IIA, MAP increased insignificantly, while in group IIB the MAP values did not change. CONCLUSIONS: Infiltration anesthesia of the projected line of skin incision in the head enabled maintenance of stability in the circulatory system and lower doses of opioids administered before commencing surgery. PMID- 11433202 TI - Liver damage in HIV-infected patients. AB - BACKGROUND: The aim of this study was to investigate the hepatic disorders in a cohort of HIV-infected patients including clinical, biochemical, serological and histopathological data. MATERIAL AND METHODS: 125 patients, 23 (18.4%) females and 102 (81.6%) males with an average age of 30.2+/-8.4 yrs, were retrospectively analysed. 61 patients (49.2%) were i. v. drug users (IVDU) and 64 patients (51.8%) were infected HIV by sexual contact. AIDS was diagnosed in 52 (41.6%) patients. RESULTS: Hepatomegaly was detected in 62 (49.6%) patients. An increased serum aspartate aminotransferase (AST) level was found in 110 (88%) patients. Testing showed 61 (48.8%) patients with evidence of a history of hepatitis B infection; 71 (56.8%) were anti-HCV positive and 40 (32%) had serological markers of both infections. 36 (28.8%) patients underwent liver biopsy. Abnormalities were detected in 32 (88%) patients. The biopsies of 13 (37.1%) patients revealed features of chronic viral hepatitis. CONCLUSIONS: HCV and HBV co-infection is frequently encountered in this cohort of HIV-infected patients. In the majority of cases the liver diseases were asymptomatic or mildly symptomatic. The hepatic disorders found in HIV-infected patients are due to different etiopathogenic factors. PMID- 11433203 TI - Parasitic cysts of the liver - practical approach to diagnosis and differentiation. AB - BACKGROUND: The aim of the study was to determine the incidence of parasitic liver cysts. MATERIAL AND METHODS: In 1996-2000 at the Department of Infectious Diseases parasitic liver cysts were diagnosed in 31 patients. The diagnosis was based on imaging examinations (ultrasound, computerised tomography), serological reactions (ELISA, IHA) and histopathological investigation of the specimens obtained through fine-needle aspiration biopsy. RESULTS AND CONCLUSION: The latter followed the pre-treatment with antiparasitic drug and no significant complications were observed. On the basis of the criteria developed by our team (evident, highly probable and probable diagnosis), hydatid disease of the liver was diagnosed in 8 patients (25.8%). The remaining subjects, excluding one patient who underwent surgical treatment, received repeated treatment with imidazole derivatives (Zentel or Vermox). PMID- 11433204 TI - Possible involvement of kynurenamines in the pathogenesis of cataract in diabetic patients. AB - BACKGROUND: It has been demonstrated that the products of tryptophan degradation, kynurenamines, play an important role in senile cataract formation. However, the involvement of these compounds in the development of diabetic cataract has not been studied. The aim of the present study was to compare the concentration of tryptophan and kynurenamines in the aqueous humor and lenses obtained from non diabetic and diabetic patients with cataract. MATERIAL AND METHODS: The concentration of tryptophan, kynurenine, 3-hydroxykynurenine, kynurenic acid and anthranilic acid was measured using high-performance liquid chromatography (HPLC) with appropriate detection in aqueous humor and lenses obtained from 38 non diabetic subjects and 20 patients with type II diabetes in course of surgical cataract extraction. RESULTS: In diabetic patients the concentration of kynurenine, 3-hydroxykynurenine and anthranilic acid in the aqueous humor was increased in comparison with non - diabetic subjects, while the concentration of tryptophan and kynurenic acid was similar in both groups. In the lenses obtained from patients with diabetes accumulation of tryptophan and all of its assayed metabolites was observed. CONCLUSIONS: Concentrations of the products of kynurenine pathway of tryptophan degradation in the aqueous humor and the lenses of diabetic patients are increased. We suggest that these compounds could play an important role in the development of diabetic cataract. PMID- 11433205 TI - Primary hepatic carcinoid tumor: case report and review of 53 cases. AB - BACKGROUND: Primary hepatic carcinoid tumor (PHCT) is a extremely rare. The authors describe a patient with PHCT and review previously published cases of the disease. CASE REPORT: A 75-year-old man, presenting with weight loss and pain in the right upper abdomen, had multiple masses in both lobes of the liver. He was diagnosed as PHCT by radiological examination, laboratory findings with high levels of 5-hydroxyindoleacetic acid (5-HIAA) in the serum and urine, and histological findings including positive staining of tumor cells for Grimelius and chromogranin A. The patient received totally transcatheter arterial chemoembolization (TACE) five times over 27 months; this treatment provided excellent palliation and caused a decrease in urinary 5-HIAA levels. Fifty-three cases of PHCT have been reported in the English-language literature. RESULTS: Analysis of these published cases revealed that PHCT occurs in the middle age (mean age = 48.2 years) and is more frequent in females (males/females = 20/33 cases). Of the symptomatic patients, the major findings is abdominal pain, fullness, and/or a palpable mass (56% of symptomatic patients). In contrast, only 2 cases out of 53 presented with symptoms of typical carcinoid syndrome. In most cases, PHCT was detected as a hypervascular lesion by radiological examination. By histological analysis, 80% and 84% of the cases were positive for Grimelius silver stain and immunohistochemically positive for chromogranin A, respectively. Surgical resection is the treatment primarily recommended with an 18% of recurrence rate and a 74% of a survival rate after 5 years. For unresectable and recurrent cases, TACE may be recommended. PMID- 11433206 TI - Pseudomembranous colitis after eradication of Helicobacter pylori infection with a triple therapy. AB - BACKGROUND: H.pylori (H.p.) infection of the gastric mucosa is causally related to chronic gastritis, peptic ulcer disease, MALT-lymphoma and gastric cancer. There is also an evidence for a link between the H.p.-infection and non-ulcer dyspepsia and even extragastric diseases. The number of patients treated against H.p. infection is expanding. Although in the last years the PPI-based triple therapies have been considered to be effective and safe, in some patients, however, severe side-effects may occur. CASE REPORT: We report on a 86 year old female patient, who received a one-week triple eradication therapy (metronidazole 3x400 mg/die, clarithromycin 2x250 mg/die and omeprazole 2x20 mg/die) because of non-ulcer dyspepsia. A few days after the eradication treatment, she developed profuse watery and bloody diarrhea and abdominal pain with distention. In stool specimens Clostridium difficile toxin was detected. A colonoscopy showed typical features of antibiotic associated pseudomembranous colitis. Until now, only few reports concerning this complication have been published and the frequency of the complication in patients eradicated for H.p. is unknown. The potential risk factors to develop this condition have not been clarified. Since the complication may be potentially lethal, this severe side-effect of the usually well-tolerated triple-therapy has to be considered in patients suffering from profuse diarrhea and abdominal pain after eradication therapy. PMID- 11433207 TI - Anomalous origin of the left coronary artery (LCA) from pulmonary trunk (Bland White-Garland syndrome) with systemic collateral supply to LCA. AB - We present the case of 15-year-old asymptomatic girl referred to our institution with the diagnosis of mitral valve prolapse and a suspicion of coronary artery fistula. Detailed diagnostics revealed Bland-White-Garland syndrome (B-W-G) with tortuous aneurysmatic right coronary artery (9 mm in diameter). In addition, on surgery, multiple collateral vessels between the right and left coronary arteries were found crossing over the pulmonary trunk and right ventricle. Because of fragile and calcified anterior walls of the main pulmonary artery we had to elect Hamilton rather than Takeuchi technique. The procedure was aggravated by continuous blood outflow from enlarged LCA ostium suggesting systemic collateral supply. Weaning from cardiopulmonary bypass and postoperative period was uneventful. There are only two published reports on systemic collateral supply to LCA in patients with BWG. PMID- 11433208 TI - Pseudoaneurysm of the gastroduodenal artery as a cause of obstructive jaundice. AB - A 35-year-old man presented to our emergency room with asymptomatic jaundice. A physical exam revealed a palpable mass with audible bruit in the epigastrium. Total serum bilirubin was 21.7 mg%. A real time sonography/Doppler examination showed widening of the biliary tree (common bile duct diameter of 13 mm) and a mass in the pancreatic head with turbulent flow. Arteriography of the celiac axis revealed a pseudoaneurysm of the gastroduodenal artery. A ligation of the gastroduodenal artery was performed surgically, and the aneurysmal cavity was explored and emptied. An intraoperative cholangiography showed slight stenosis of the common bile duct distally, and so a choledochojejunostomy was performed. The patient's recovery was uneventful. A follow-up angiogram revealed the short stump of the gastroduodenal artery and no aneurysm or extravasation of dye. A follow-up ultrasound showed the common bile duct measuring 5.5 mm. The bilirubin level dropped to normal values. The patient was discharged on 12 days after surgery. Ten months following surgery he was doing well. The pathology, diagnosis, and treatment of such cases are briefly discussed. PMID- 11433209 TI - MR proton spectroscopy in patients with CNS involvement in Bourneville's disease. AB - BACKGROUND: Despite much previous research, the origin of intracranial lesions in tuberous sclerosis is still unknown. We present MR and proton magnetic resonance (1H-MRS) studies of two patients with tuberous sclerosis. MATERIAL AND METHODS: The study material consisted of MR and 1H-MRS examinations from 2 patients (4-5 years old) with tuberous sclerosis. MR images (FSE, FAST, FLAIR) were obtained with a 1.5 T unit. Localized 1H-MRS studies (PRESS 35, TE 35 ms, TR 1500 ms, NEX 192) on cortical (n=3) and subependymal (n=1) lesions were also performed. RESULTS: Decreased N-acatyloaspartate/creatine (NAA/Cr) and increased myoinositol/creatine (mI/Cr) ratios were observed in the spectra obtained from the subcortical and subependymal tubers. CONCLUSIONS: Our observations confirm the hypothesis that disturbances of migration, adhesion and differentiation of germ neuronal cells may underlay the pathogenesis of tuberous sclerosis. PMID- 11433210 TI - Application of intraoperative thermography in coronary artery bypass grafting without extracorporeal circulation - clinical report. AB - BACKGROUND: The development of low-invasive cardiosurgical techniques has contributed to an increase of the number of revascularization procedures performed on beating heart. The report presents the changes observed intraoperatively in thermographic images of the heart obtained during surgical revascularization performed without extracorporeal circulation. MATERIAL AND METHODS: In two patients with coronary heart disease, subjected to coronary artery bypass grafting performed without extracorporeal circulation, the images acquired over the operation area were recorded intraoperatively using a thermovision camera (Agema Thermovision SW/TE 900 system). RESULTS: On the basis of thermographic images, the area of lowered temperature corresponding to the region of myocardium with impaired blood flow supplied by the descending anterior coronary artery branch was determined. A rapid temperature increase to the baseline values was observed soon after removing the ligatures from the descending anterior coronary artery and left internal mammary artery used as a bypass. CONCLUSIONS: Recording thermographic heart images allows to observe restricted areas of perfusion disturbances, which take place during procedures performed on beating heart. PMID- 11433211 TI - Why Congo red binding is specific for amyloid proteins - model studies and a computer analysis approach. AB - BACKGROUND: The complexing of Congo red in two different ligand forms - unimolecular and supramolecular (seven molecules in a micelle) - with eight deca peptides organized in a b-sheet was tested by computational analysis to identify its dye-binding preferences. Polyphenylananine and polylysine peptides were selected to represent the specific side chain interactions expected to ensure particularly the stabilization of the dye-protein complex. Polyalanine was used to verify the participation of non-specific backbone-derived interactions. MATERIAL AND METHODS: The initial complexes for calculation were constructed by intercalating the dye between the peptides in the middle of the beta-sheet. The long axis of the dye molecule (in the case of unimolecular systems) or the long axis of the ribbon-like micelle (in the case of the supramolecular dye form) was oriented parallel to the peptide backbone. This positioning maximally reduced the exposure of the hydrophobic diphenyl (central dye fragment) to water. In general the complexes of supramolecular Congo red ligands appeared more stable than those formed by individual dye molecules. Specific interactions (electrostatic and/or ring stacking) dominated as binding forces in the case of the single molecule, while non-specific surface adsorption seemed decisive in complexing with the supramolecular ligand. RESULTS: Both the unimolecular and supramolecular versions of the dye ligand were found to be likely to form complexes of sufficient stability with peptides. The low stability of the protein and the gap accessible to penetration in the peptide sheet seem sufficient for supramolecular ligand binding, but the presence of positively charged or hydrophobic amino acids may strengthen binding significantly. CONCLUSIONS: The need for specific interaction makes single-molecule Congo red binding rather unusual as a general amyloid protein ligand. The structural feature of Congo red, which enables specific and common interaction with amyloid proteins, probably derives from the ribbon-like self-assembled form of the dye. PMID- 11433212 TI - Soluble thrombomodulin: a marker of endothelial injury, an antithrombotic agent, or both? PMID- 11433213 TI - Menarini Academy Cardiovascular Research Awards in Basic Science 2001: ligands of the orphan receptor peroxisome-proliferator activator-gamma reduce myocardial infarct size. PMID- 11433214 TI - My life and times with enzymes and mediators. AB - The vascular endothelium plays a vital role in the control of the circulation. It metabolises various vasoactive substances, converts angiotensin I to angiotensin II and secretes the potent vasodilator prostacyclin, as well as nitric oxide and endothelin-1. The balance between these mediators determines the response of the cardiovascular system in diseases such as hypertension, atherosclerosis and myocardial infarction. In this article I describe some of my work over the past 50 years. In the process, I hope to show that it is not only Princes who travel the road to Serendip but also Scientists. The application of simple methods, luck, happy coincidences, serendipity and noticing the unusual have all played an important role in my scientific career. However, the choice of colleagues has also been vital. PMID- 11433215 TI - NO, nitrotyrosine, and cyclic GMP in signal transduction. AB - Over the past 25 years, the role of nitric oxide (NO) in biology has evolved from being recognized as an environmental pollutant to an endogenously produced substance involved in cell communication and signal transduction. NO is produced by a family of enzymes called nitric oxide synthases (NOSs), which can be stimulated by a variety of factors that mediate responses to various stimuli. NO can initiate its biological effects through activation of the heterodimeric enzyme, soluble guanylyl cyclase (sGC), or through several other chemical reactions. Activation of sGC results in the production of 3',5'-cyclic guanosine monophosphate (cGMP), an intracellular second messenger signaling molecule, which can subsequently mediate such diverse physiological events such as vasodilatation and immunomodulation. Chemically reactive NO can affect physiological changes through modifications to cellular proteins, one of which is tyrosine nitration. The demonstration that NO is involved in so many biological pathways indicates the importance of this endogenously produced substance, and suggests that there is much more to be discovered about its role in biology in years to come. PMID- 11433216 TI - Connective tissue skeleton in the normal left ventricle and in hypertensive left ventricular hypertrophy and chronic chagasic myocarditis. AB - Pictures certainly are worth a thousand words in the case of the structure of the connective tissue skeleton of normal and diseased myocardium. This report reviews the connective tissue matrix of the normal human myocardial tissue and the pathological myocardial fibrosis in left ventricular hypertrophy due to chronic arterial hypertension in humans and in human chronic chagasic myocarditis. The myocardial connective tissue matrix was studied employing a cell-maceration method that removes the myocardial tissue non-fibrous elements, and leaves behind a non-collapsed matrix, thus allowing a better three-dimensional view. Such information extends our knowledge of the expression of interstitial myocardial fibrous tissue in normal hearts and in hypertensive left ventricular hypertrophy and chronic chagasic myocarditis. The progressive accumulation of interstitial collagen fibers in both chronic cardiac diseases may be expected to decrease myocardial compliance and disrupt synchronous contractions of the ventricles during systole, contributing to a spectrum of ventricular dysfunction that involve either the diastolic or systolic phase of the cardiac cycle or both. In hypertensive heart disease myocardial fibrosis can be also implicated in the genesis of ventricular dysrhythmias, possible causes of sudden death among chronic hypertensive patients. Regarding chronic chagasic myocarditis, myocardial fibrosis is probably implicated in the genesis of malignant ventricular tachyarrhythmias (ventricular tachycardia and ventricular fibrillation), major causes of sudden death among patients with chronic Chagas' heart disease. The collagen distribution could interfere on the electrical properties of the myocardium. Fibrosis can block the cardiac impulse that may recycle (re-entry) through an alternative route and could slow conduction. In addition, the thick collagenous septa encompassing muscle fiber bundles could interfere with lateral impulse conduction, which would favor re-entry. Moreover, the methodology used is a useful tool to study the spatial organization of the collagen fibrils of the myocardium under normal and pathological conditions. PMID- 11433217 TI - Syringe, pen, inhaler - the evolution of insulin therapy. AB - The therapy with inhalable insulin can be expected to change the present concepts and the market of subcutaneous insulin dramatically within the next years. Several companies are currently developing formulations of inhalable insulin. In the most advanced concept, insulin is delivered as a dry-powder insulin formulation via a special aerosol device system. The phase III studies evaluating the efficiency of the inhaled insulin are already on their way. The recent phase II studies have shown, that the lung is capable of absorbing new insulin formulation in a dose-dependent and reproducible manner. However, a relatively small number of diabetic patients have been included in these studies, yet. The therapeutic efficacy and safety of the inhaled insulin is comparable to that of the usual subcutaneous insulin treatment regimens. The most important advantage of the new therapy is the enhanced therapeutic comfort of the patient who does not need to inject insulin for meal time glucose control. Generally, in terms of glycemic control, inhalable insulin offers no advantages in type 1 diabetics in comparison to an intensified conventional insulin therapy. However, before a large-scale marketing, several open questions have to be carefully investigated, the most important being the possible long-term effects of insulin inhalation for the lung, since insulin is known to have growth-promoting properties. There is still no available clinical data concerning the efficiency of the inhaled insulin in patients with pulmonary diseases which may cause problems in absorption of inhaled insulin due to the smaller cumulative alveolar surface. In smokers without pulmonary disease seems the inhaled insulin to act stronger and faster. Since therapy with inhalable insulin requires larger doses of insulin in comparison to subcutaneous insulin to achieve the same systemic effect, the costs of this therapy need to be clarified, too. PMID- 11433218 TI - A pleiotropic antiatherogenic action of ibuprofen. AB - Ibuprofen is a cyclooxygenase (COX-1 and COX-2) inhibitor known to reduce the production of prostaglandins that play prominent role in inflammation. Other properties of the drug, aside from its anti-inflammatory effects, have been recently studied. In this paper we shall discuss several properties of ibuprofen that making the drug interesting for treatment of conditions associated with atherosclerosis. Ibuprofen exerts pleiotropic effects such as inhibition of adhesion and transendothelial migration of leukocytes, suppressing intracellular production of reactive oxygen species and oxidative modification of LDL. Interestingly, ibuprofen increased HDL cholesterol levels and reduced the level of triglicerides. Ibuprofen can also modulate efficiency of fibrynolisis by inhibiting production of plasminogen activator inhibitor (PAI-1). This properties of ibuprofen may be due to changing the activity of transcription factors. Ibuprofen inhibits the activation of NF-kB and activates PPARa and PPARg. PMID- 11433219 TI - Bonum vinum laetificat cor hominum. AB - Beneficial effects of wine consumption on health have been suspected since the antiquity. Recent epidemiological studies show that coronary heart disease mortality markedly decreases from northern to southern Europe and is lower in Mediterranean than in other developed countries. Because wine is a component of the Mediterranean diet, it has been suggested that moderate wine especially red wine consumption may produce additional beneficial effects on cardiovascular morbidity and mortality compared to consuming the same quantity of alcohol in other beverages. Polyphenols are good candidates to explain the putative cardiovascular protective effect of wine, because they are abundant in wine especially red wine, and possess antioxidant and superoxide ion scavenging properties. Because it is readily accessible from blood and produces cardioprotective agents like nitric oxide (NO) the endothelial cell may be a privileged target for wine polyphenols. Polyphenols from red wine can prevent oxidation of low density lipoproteins (LDL). As oxidized LDL inhibit agonist activated NO release from endothelial cells and subsequent endothelium-dependent relaxation of arteries, wine polyphenols might prevent LDL-induced alterations of endothelial function. Furthermore some wine polyphenols contained in oligomeric condensed tannins- and anthocyaninsD enriched fractions can act directly on endothelial cells to cause calcium-dependent release of NO. The latter effect is independent from superoxide scavenging and antioxidant properties of the polyphenols, and it is produced by compounds with specific structures only. Thus, decreased oxidation of LDL and enhanced release of NO from endothelium caused by polyphenols from red wine may result in cardiovascular protection. However further studies are required to demonstrate whether or not these effects are involved in the putative protective effect of wine on cardiovascular morbidity and mortality. PMID- 11433220 TI - How recombinant insulin analogs improve insulin therapy of diabetes mellitus: pathophysiology, clinical practice and recommendations. AB - According to the latest prognoses, 300 million people are expected to suffer from diabetes (particularly type 2) in 2025; diabetes will become a new epidemics of the coming century. Therefore it is so important to get acquainted with the pathomechanism of diabetes-related disorders and the possibilities of effective treatment. During the last decade, numerous prospective epidemiological studies have been completed, which have considerably extended our knowledge of the pathomechanism of disturbances and their effect on late complications of diabetes. Resistance to insulin has an important role in the etiology of type 2 diabetes. However, there is substantial evidence indicating that impairment or regression of rapid insulin secretion phase at the early stage of the disease is a factor of no lesser importance. The lack of appropriate insulin concentration after the stimulation by food ingestion (first 30 min after the meal), leads to permanent, treatment-resistant hyperglycemia. The postulated attempts to obtain a 'nearly normoglycemic' condition, as well as the results of prospective epidemiological studies proving that achieving this aim improves the prognosis of diabetic patients, prompt the researchers to search for the drugs or treatment schedules, which could effectively restore the impaired insulin secretion, and its early phase in particular, in type 2 diabetes. Human insulin analogs and their pre-formulated commercially available mixtures seem very promising in this respect. PMID- 11433221 TI - Treatment of rheumatoid arthritis: Tightening the noose. PMID- 11433222 TI - Treatment of hyponatremic encephalopathy with antagonists to antidiuretic hormone. PMID- 11433223 TI - New disease-modifying antirheumatic drug 2 acetylthiomethyl-4-(4-methylphenyl)-4 oxobutanoic acid (KE-298) selectively augments activation-induced T cell death. AB - We examined in this study whether the newly developed disease-modifying antirheumatic drug (DMARD) 2-acetylthiomethyl-4-(4-methylphenyl)-4-oxobutanoic acid (KE-298) augments activation-induced T cell death. Peripheral blood (PB) T cells, isolated from healthy donors, were activated by incubation with interleukin-2 (IL-2) followed by further culture with 12-0-tetradecanoyl phorbol 13-acetate (PMA) and ionomycin in the presence or absence of KE-298. The apoptosis of activated T cells was examined by flow cytometric determination of hypodiploid DNA. Fas expression and caspase-3 activity in activated T cells were also examined by flow cytometry, and expression of Fas ligand (FasL), Bcl-2 related proteins, and X chromosome-linked inhibitor of apoptosis protein (XIAP) was determined by Western blot analysis. Apoptosis was not obvious in resting T cells and was not augmented by KE-298. In contrast, apoptosis was clearly detected in activated T cells (activation-induced T cell death) with the increment of caspase-3 activity, and incubation of these cells with KE-298 further enhanced apoptosis. Treatment of activated T cells with KE-298 increased Bax expression but decreased XIAP expression without affecting the expression of Fas/FasL. Thus caspase-3 activity in activated T cells appeared to be increased by KE-298. Our results suggest that the newly developed DMARD, KE-298, selectively augmented activation-induced T cell death. This finding may contribute to the therapeutic efficacy of KE-298 in rheumatoid arthritis (RA) patients and provide new insight into the pharmacologic action of DMARDs. PMID- 11433224 TI - Difference in solute excretion during correction of hyponatremic patients with cirrhosis or syndrome of inappropriate secretion of antidiuretic hormone by oral vasopressin V2 receptor antagonist VPA-985. AB - VPA-985 is an orally active, competitive vasopressin V(2) receptor antagonist that in normal human beings increases water excretion without affecting solute excretion. Whether solute excretion is affected in patients with hyponatremia resulting from inappropriate secretion of antidiuretic hormone (SIADH) or from cirrhosis treated with VPA-985 is unknown. Six hyponatremic patients with SIADH and 5 hyponatremic patients with cirrhosis with ascitis (CWAs) were treated with 50 or 100 mg VPA-985 twice daily. Evolution of creatinine, urea, uric acid, sodium, potassium, and osmotic clearance were determined. Volume hormones (plasma renin [PR], aldosterone, antidiuretic hormone [ADH], atrial natriuretic factor [ANF]) were also determined before and after treatment. In patients with SIADH, serum sodium concentration (SNa) was generally corrected in 1 day (SNa: 126 +/- 4.5 mmol/L at t = 0 hours and 133 +/- 5.6 mmol/L at t = 24 hours) and associated with a decrease in sodium excretion (from 82 +/- 22 mmol/24 hours to 45 +/- 21 mmol/24 hours; P < 0.05) without modification in potassium excretion. Despite an increase in diuresis (from 0.84 +/- 0.2 ml/min to 1.46 +/- 0.4 ml/min) urea and uric acid clearances decreased. Urine osmolality decreased from 414 +/- 148 mOsm/kg H(2)O to 209 +/- 55 mOsm/kg H(2)O. Volume hormones did not change. In the CWAs the rise of SNa was more progressive (SNa: 126 +/- 2.8 mmol/L at t = H0 to 133 +/- 4.9 mmol/L at t = 48 hours) and parallel to an augmentation in sodium excretion (from 23 +/- 18 mmol/24 hours to 65 6 60 mmol/24 hours the second day of VPA administration). The higher sodium excretion was also connected with a progression in potassium excretion (from 22 6 7 mmol/24 hours to 36 +/- 18 mmol/24 hours). The increase in diuresis under VPA from 0.42 +/- 0.2 mL/min to 1.7 +/- 0.9 mL/min resulted in a higher urea clearance. Urine osmolality decreased from 509 +/- 142 mOsm/kg H(2)O before VPA to 194 +/- 106 mOsm/kg H(2)O after VPA. ADH increased in CWAs treated with VPA, from 1.9 +/- 1.2 pg/mL to 5.3 +/- 2.8 pg/mL (P <.05) while other volume hormones did not change. VPA-985 is a highly effective drug in the short-term management of hyponatremic patients with SIADH or CWAs. SNa correction is associated with urinary sodium retention in SIADH, whereas in CWAs a mild increase in sodium excretion is observed. PMID- 11433225 TI - Heparin is not required for detection of antibodies associated with heparin induced thrombocytopenia/thrombosis. AB - Heparin-induced thrombocytopenia (HIT), with or without thrombosis, is a common and often serious complication of heparin therapy. Platelet-activating, heparin induced antibodies characteristic of HIT are thought to be specific for complexes formed between platelet factor 4 (PF4) and heparin, and such complexes are routinely used for antibody detection. We studied the binding of HIT antibodies to PF4 complexed with heparin fractions of uniform molecular size or linear polyanions other than heparin and found that many compounds other than heparin form complexes with PF4 that are suitable for antibody detection, provided they carry strong negative charges spaced about 0.5 nm apart along the molecular backbone and are of sufficient length to span about 40% of the circumference of the PF4 tetramer. Polyvinyl phosphonate was among the compounds that were equivalent to heparin. Thus neither a polysaccharide chain nor sulfate side groups--the hallmarks of heparin structure--are required for HIT antibody detection. The findings support the view that antibodies associated with HIT are specific for conformational changes that take place in the positively charged PF4 molecule when it reacts with a suitable, linear polyanion. PMID- 11433226 TI - Secoisolariciresinol diglucoside from flaxseed delays the development of type 2 diabetes in Zucker rat. AB - Previous research has suggested that type 1 diabetes mellitus may be due to oxidative stress. The role of oxidative stress in type 2 diabetes is not known. Secoisolariciresinol diglucoside (SDG) antioxidant, obtained from flaxseed, has been reported to prevent type 1 diabetes in a rat model. However, its effectiveness in type 2 diabetes is not known. An investigation was made of the effects of SDG isolated from flaxseed (40 mg/kg body wt, orally in drinking water) on the development of diabetes in Zucker diabetic fatty (ZDF)/Gmi-fa/fa female rats, a model of human type 2 diabetes, to determine whether this type of diabetes is due to oxidative stress and whether SDG could prevent the development of diabetes. A total of 10 Zucker lean control and 26 ZDF rats were used in this study. Incidence of diabetes was 100% in untreated and 20% in SDG-treated ZDF rats by the age of 72 days (P <.01). The rats that did not develop diabetes by 72 days of age in the SDG-treated group developed diabetes later on (age 72 to 99 days) except for 10% of the rats that did not develop diabetes for the duration of the study (101 days of age), suggesting that SDG retarded the development of diabetes. Diabetes was associated with an increase in oxidative stress as suggested by an increase in serum malondialdehyde (P <.01). Also, diabetes was associated with an increase in serum total cholesterol and triglycerides (P <.05) and glycated hemoglobin A(1C) (P <.05). ZDF rats treated with SDG that did not develop diabetes by 70 days of age had no increase in oxidative stress, serum total cholesterol, and glycated hemoglobin. These results suggest that type 2 diabetes is associated with an increase in oxidative stress and that SDG is effective in retarding the development of diabetes. PMID- 11433227 TI - Both subtype 1 and 2 receptors of angiotensin II participate in regulation of intracellular calcium in glomerular epithelial cells. AB - We have documented that both receptors of angiotensin II (ANG II) (AT1 and AT2) are involved in regulation of intracellular signals in glomerular epithelial cells (GECs). We studied the role of these receptors in regulation of intracellular ionized calcium [Ca2(+)]i in GECs. Cells were loaded with Indo-1 (Ca2(+)) and SNARF-1 (pH) fluorescent dyes and then incubated with or without ANG II for 1 hour at 37 degrees C. In some experiments AT(1) and AT(2) receptor blockers (Losartan and PD 12339, respectively) were added. In additional experiments cells were incubated with thapsigargin (Tg) and bradykinin (BK) as well as ANG II. A four-channel fluorescence videomicroscope system was used to measure real-time [Ca2(+) ]i in individual cells. Levels of inositol triphosphate (IP(3)) were measured with radioimmunoassay. An amount of 100 nmol/L of ANG II caused a maximal increase in [Ca2(+)]i in calcium-containing buffer. ANG II had no effect on intracellular pH of GECs. Increase in [Ca2(+)]i by ANG II was prevented by the concurrent use of Losartan and PD 123319. BK caused a transient increase in [Ca2(+)]i, which was significantly decreased by ANG II; concurrent addition of Losartan and PD 123319 prevented ANG II effect. ANG II prevented the accumulation of Ca2(+) in intracellular stores. ANG II caused a significant but transient increase in levels of IP(3). In summary, ANG II increases extracellular/intracellular calcium dependent bidirectional Ca2(+) transport in GECs, inhibits BK induced release of Ca2(+) from IP(3) sensitive stores, and, in addition, reduces refilling of endoplasmic reticulum [Ca2(+)] depleted by repeated BK stimulation. Both receptor subtypes appear to be important in ANG II mediated physiologic responses of GECs and may participate in modulation of glomerular function in vivo. PMID- 11433228 TI - Contribution of nitric oxide to the protective effects of ischemic preconditioning in ischemia-reperfused rat kidneys. AB - We examined the contribution of nitric oxide (NO) to the effect of ischemic preconditioning (IP) on renal function and the hemodynamics in ischemia reperfusion (I/R) mediated kidney injury. IP was performed by using 4 minutes of ischemia followed by a 30-minute reperfusion interval. I/R treatment consisted of a 30-minute ischemia and 60-minute reperfusion interval. We measured the glomerular filtration rate (GFR), the fractional excretion of sodium (FE(Na)), and the renal blood flow (RBF) in IP+I/R and I/R kidneys. Rats were pretreated with NaCl, N(G)-nitro-L-arginine methyl ester (L-NAME), or L-arginine. We found that IP significantly improved GFR and FE(Na) as compared with I/R treatment; however, this effect was completely abolished by L-NAME injection and enhanced by L-arginine treatment. L-NAME treatment significantly diminished RBF but did not alter nitrite/nitrate excretion. Furthermore, we found that IP alone does not lead to inducible NO synthase protein expression whereas I/R or IP+I/R treatment clearly did. Moreover, we observed an increased heme oxygenase-1 expression in IP+I/R kidneys as compared with I/R treated ones. Our results clearly showed that IP pretreatment protects kidneys from I/R mediated tissue injury and that these effects were partially mediated by NO. PMID- 11433229 TI - Advanced glycosylation end products stimulate collagen mRNA synthesis in mesangial cells mediated by protein kinase C and transforming growth factor-beta. AB - Advanced glycosylation end products (AGE) seem to be implicated in the pathogenesis of diabetic nephropathy. The present study has examined the effects of AGE on protein kinase C (PKC) activity and transforming growth factor-beta1 (TGF-beta1) in relation to collagen gene regulation in cultured human mesangial cells (HMCs). Quiescent HMCs were exposed to serum-free media containing bovine serum albumin (BSA), AGE-modified BSA (AGE-BSA), or glycated BSA in which AGE formation was prevented by the use of aminoguanidine (BSA-AM). AGE-BSA (200 microg/mL) induced a peak membrane-associated PKC activity, particularly PKC-a, at 4 hours. AGE-BSA stimulated alpha1(I) and alpha1(IV) collagen mRNA expression after 24-hour incubation with HMCs, which remained elevated until hour 60. HMCs incubated with AGE-BSA induced a significant inhibition of cell proliferation compared with cells incubated with BSA. AGE-BSA stimulated TGF-beta mRNA and protein expression in HMCs. The TGF-beta secreted by HMCs was shown by CCL-64 mink lung cell assay to be bioactive. In contrast, BSA-AM did not affect either collagen or TGF-beta mRNA or protein expression in HMCs. The stimulatory effects of AGE-BSA on collagen gene regulation in HMCs could be negated by the pretreatment of HMCs with GF 109203X for 30 minutes or with phorbol myristate acetate for 24 hours before AGE-BSA administration. Neutralizing antibody to TGF beta inhibited increased collagen mRNA expression by HMCs exposed to AGE-BSA. These results suggest that AGE-BSA stimulates collagen mRNA expression by activating PKC and the transcriptional upregulation of TGF-beta1 in HMCs. Thus, PKC and TGF-beta may function as key signaling intermediaries in the AGE-up regulated collagen gene expression pathway in HMCs. PMID- 11433230 TI - Mycophenolate mofetil reduces renal cortical inducible nitric oxide synthase mRNA expression and diminishes glomerulosclerosis in MRL/lpr mice. AB - Overexpression of inducible nitric oxide synthase (iNOS) has been implicated in the pathogenesis of lupus glomerulonephritis. Mycophenolate mofetil (MMF), a novel immunosuppressive agent, is currently used in organ transplantation and under evaluation for treatment of autoimmune disorders. Mycophenolic acid, the active metabolite of MMF, has been shown to suppress cytokine-induced nitric oxide production in vitro. The aim of this study was to evaluate the effect of MMF on the expression of renal cortical iNOS mRNA and protection against glomerulonephritis in MRL/lpr mice. Three-month-old MRL/lpr mice (n = 6) displaying clinical symptoms of glomerulonephritis were treated for 3 months with MMF (90 mg/kg/day) dissolved in a vehicle. Controls were age- and sex-matched mice (n = 6) that received the vehicle alone. By reverse-transcription competitive polymerase chain reaction, we found that the renal cortical iNOS/beta actin mRNA ratio was reduced by 30.8% (P <.05) in MMF-treated mice. Furthermore, MMF significantly reduced urinary nitrite production and degree of glomerulosclerosis. The glomerular volume was reduced by 17.5% (P <.001). Proteinuria was also significantly reduced in the MMF-treated group. However, by electrophoretic mobility shift assay, the nuclear binding of nuclear factor kappaB (NF-kappaB) was not affected by MMF treatment. We conclude that in addition to its immunosuppressive action, MMF may reduce renal cortical iNOS mRNA expression and diminish glomerulosclerosis in MRL/lpr mice independent of modulation of the NF-kappaB pathway. PMID- 11433233 TI - Creating a winning patent portfolio. PMID- 11433243 TI - Evolving paradigms in biotech IPO valuations. PMID- 11433244 TI - Perfect partnering. PMID- 11433245 TI - Managing business risk. PMID- 11433246 TI - Clinical development--look before you leap. PMID- 11433247 TI - Just human nature. PMID- 11433248 TI - Good relations--PR for the biotech business. PMID- 11433249 TI - When products fail. PMID- 11433250 TI - Information technology to the rescue! PMID- 11433251 TI - Monopoly money. PMID- 11433252 TI - The headache of knowledge management. PMID- 11433253 TI - Differentiation and integration. PMID- 11433254 TI - Approval heralds new generation of kinase inhibitors? PMID- 11433255 TI - Sequenom steps toward drug development. PMID- 11433256 TI - South Africa seeks genomic partners. PMID- 11433257 TI - Disease genes less elusive than once thought, say researchers. PMID- 11433258 TI - Fake biotech drugs raise concerns. PMID- 11433259 TI - Italian public votes out anti-GMO Greens. PMID- 11433260 TI - Gene pill patents a surprise for gene therapy. PMID- 11433262 TI - Real and metaphorical moral limits in the biotech debate. PMID- 11433264 TI - GMO contamination of seeds. PMID- 11433267 TI - Making a living out of the art of dying. PMID- 11433268 TI - Bar-coding biomolecules with fluorescent nanocrystals. PMID- 11433269 TI - Nanopores with a spark for single-molecule detection. PMID- 11433270 TI - Crafting crop resistance to corn rootworms. PMID- 11433271 TI - The path not taken. PMID- 11433273 TI - Quantum-dot-tagged microbeads for multiplexed optical coding of biomolecules. AB - Multicolor optical coding for biological assays has been achieved by embedding different-sized quantum dots (zinc sulfide-capped cadmium selenide nanocrystals) into polymeric microbeads at precisely controlled ratios. Their novel optical properties (e.g., size-tunable emission and simultaneous excitation) render these highly luminescent quantum dots (QDs) ideal fluorophores for wavelength-and intensity multiplexing. The use of 10 intensity levels and 6 colors could theoretically code one million nucleic acid or protein sequences. Imaging and spectroscopic measurements indicate that the QD-tagged beads are highly uniform and reproducible, yielding bead identification accuracies as high as 99.99% under favorable conditions. DNA hybridization studies demonstrate that the coding and target signals can be simultaneously read at the single-bead level. This spectral coding technology is expected to open new opportunities in gene expression studies, high-throughput screening, and medical diagnostics. PMID- 11433274 TI - Sequence-specific detection of individual DNA strands using engineered nanopores. AB - We describe biosensor elements that are capable of identifying individual DNA strands with single-base resolution. Each biosensor element consists of an individual DNA oligonucleotide covalently attached within the lumen of the alpha hemolysin (alphaHL) pore to form a "DNA-nanopore". The binding of single-stranded DNA (ssDNA) molecules to the tethered DNA strand causes changes in the ionic current flowing through a nanopore. On the basis of DNA duplex lifetimes, the DNA nanopores are able to discriminate between individual DNA strands up to 30 nucleotides in length differing by a single base substitution. This was exemplified by the detection of a drug resistance-conferring mutation in the reverse transcriptase gene of HIV. In addition, the approach was used to sequence a complete codon in an individual DNA strand tethered to a nanopore. PMID- 11433275 TI - The HIV Tat protein transduction domain improves the biodistribution of beta glucuronidase expressed from recombinant viral vectors. AB - Treatment of inherited genetic diseases of the brain remains an intractable problem. Methods to improve the distribution of enzymes that are injected or expressed from transduced cells will be required for many human brain therapies. Recent studies showed that a peptide, the protein transduction domain (PTD) from HIV Tat, could improve the distribution of cytoplasmic reporter proteins when administered systemically as fusion proteins or cross-linked chimeras. The utility of this motif for noncytoplasmic proteins has not been determined. Here, we tested how the Tat motif affected uptake and biodistribution of the lysosomal enzyme beta-glucuronidase, the protein deficient in the disease mucopolysaccharidosis VII, when expressed from viral vectors. The Tat motif allowed for mannose-6-phosphate (M6P) independent uptake in vitro and significantly increased the distribution of beta-glucuronidase secreted from transduced cells after intravenous or direct brain injection in mice of recombinant vectors. Thus, enzymes modified to contain protein transduction motifs may represent a general strategy for improving the distribution of secreted proteins following in vivo gene transfer. PMID- 11433276 TI - Multiphoton-evoked color change of DsRed as an optical highlighter for cellular and subcellular labeling. AB - DsRed, a recently cloned red fluorescent protein, has attracted great interest as an expression tracer and fusion partner for multicolor imaging. We report that three-photon excitation (lambda <760 nm) rapidly changes the fluorescence of DsRed from red to green when viewed subsequently by conventional (one-photon) epifluorescence. Mechanistically, three-photon excitation (lambda <760 nm) selectively bleaches the mature, red-emitting form of DsRed, thereby enhancing emission from the immature green form through reduction of fluorescence resonance energy transfer (FRET). The "greening" effect occurs in live mammalian cells at the cellular and subcellular levels, and the resultant color change persists for >30 h without affecting cell viability. This technique allows individual cells, organelles, and fusion proteins to be optically marked and has potential utility for studying cell lineage, organelle dynamics, and protein trafficking, as well as for selective retrieval of cells from a population. We describe optimal parameters to induce the color change of DsRed, and demonstrate applications that show the potential of this optical highlighter. PMID- 11433277 TI - In vitro selection of nucleoprotein enzymes. AB - Natural nucleic acids frequently rely on proteins for stabilization or catalytic activity. In contrast, nucleic acids selected in vitro can catalyze a wide range of reactions even in the absence of proteins. To augment selected nucleic acids with protein functionalities, we have developed a technique for the selection of protein-dependent ribozyme ligases. After randomizing a previously selected ribozyme ligase, L1, we selected variants that required one of two protein cofactors, a tyrosyl transfer RNA (tRNA) synthetase (Cyt18) or hen egg white lysozyme. The resulting nucleoprotein enzymes were activated several thousand fold by their cognate protein effectors, and could specifically recognize the structures of the native proteins. Protein-dependent ribozymes can potentially be adapted to novel assays for detecting target proteins, and the selection method's generality may allow the high-throughput identification of ribozymes capable of recognizing a sizable fraction of a proteome. PMID- 11433278 TI - A rapid, generally applicable method to engineer zinc fingers illustrated by targeting the HIV-1 promoter. AB - DNA-binding domains with predetermined sequence specificity are engineered by selection of zinc finger modules using phage display, allowing the construction of customized transcription factors. Despite remarkable progress in this field, the available protein-engineering methods are deficient in many respects, thus hampering the applicability of the technique. Here we present a rapid and convenient method that can be used to design zinc finger proteins against a variety of DNA-binding sites. This is based on a pair of pre-made zinc finger phage-display libraries, which are used in parallel to select two DNA-binding domains each of which recognizes given 5 base pair sequences, and whose products are recombined to produce a single protein that recognizes a composite (9 base pair) site of predefined sequence. Engineering using this system can be completed in less than two weeks and yields proteins that bind sequence-specifically to DNA with Kd values in the nanomolar range. To illustrate the technique, we have selected seven different proteins to bind various regions of the human immunodeficiency virus 1 (HIV-1) promoter. PMID- 11433279 TI - Determination of protease cleavage site motifs using mixture-based oriented peptide libraries. AB - The number of known proteases is increasing at a tremendous rate as a consequence of genome sequencing projects. Although one can guess at the functions of these novel enzymes by considering sequence homology to known proteases, there is a need for new tools to rapidly provide functional information on large numbers of proteins. We describe a method for determining the cleavage site specificity of proteolytic enzymes that involves pooled sequencing of peptide library mixtures. The method was used to determine cleavage site motifs for six enzymes in the matrix metalloprotease (MMP) family. The results were validated by comparison with previous literature and by analyzing the cleavage of individually synthesized peptide substrates. The library data led us to identify the proteoglycan neurocan as a novel MMP-2 substrate. Our results indicate that a small set of libraries can be used to quickly profile an expanding protease family, providing information applicable to the design of inhibitors and to the identification of protein substrates. PMID- 11433280 TI - Insecticidal proteins from Bacillus thuringiensis protect corn from corn rootworms. AB - Field tests of corn co-expressing two new delta-endotoxins from Bacillus thuringiensis (Bt) have demonstrated protection from root damage by western corn rootworm (Diabrotica virgifera virgifera LeConte). The level of protection exceeds that provided by chemical insecticides. In the bacterium, these proteins form crystals during the sporulation phase of the growth cycle, are encoded by a single operon, and have molecular masses of 14 kDa and 44 kDa. Corn rootworm larvae fed on corn roots expressing the proteins showed histopathological symptoms in the midgut epithelium. PMID- 11433281 TI - An invasive cleavage assay for direct quantitation of specific RNAs. AB - RNA quantitation is becoming increasingly important in basic, pharmaceutical, and clinical research. For example, quantitation of viral RNAs can predict disease progression and therapeutic efficacy. Likewise, gene expression analysis of diseased versus normal, or untreated versus treated, tissue can identify relevant biological responses or assess the effects of pharmacological agents. As the focus of the Human Genome Project moves toward gene expression analysis, the field will require a flexible RNA analysis technology that can quantitatively monitor multiple forms of alternatively transcribed and/or processed RNAs (refs 3,4). We have applied the principles of invasive cleavage and engineered an improved 5'-nuclease to develop an isothermal, fluorescence resonance energy transfer (FRET)-based signal amplification method for detecting RNA in both total RNA and cell lysate samples. This detection format, termed the RNA invasive cleavage assay, obviates the need for target amplification or additional enzymatic signal enhancement. In this report, we describe the assay and present data demonstrating its capabilities for sensitive (<100 copies per reaction), specific (discrimination of 95% homologous sequences, 1 in > or =20,000), and quantitative (1.2-fold changes in RNA levels) detection of unamplified RNA in both single- and biplex-reaction formats. PMID- 11433282 TI - Bidirectionalization of polar promoters in plants. AB - A typical eukaryotic promoter consists of a minimal promoter and other upstream cis elements. The minimal promoter is essentially a TATA box region where RNA polymerase II, TATA-binding protein (TBP), and TBP-associated factors (TAFs) bind to initiate transcription, but minimal promoters alone have no transcriptional activity. The cis elements, to which tissue-specific or development-specific transcription factors bind, individually or in combination, determine the spatio temporal expression pattern of a promoter at the transcriptional level. The arrangement of upstream cis elements followed by a minimal promoter sets the polarity of the promoter. Promoters in plants that have been cloned and widely used for both basic research and biotechnological application are generally unidirectional, directing only one gene that has been fused at its 3' end (downstream). It is often necessary to introduce multiple genes into plants for metabolic engineering and trait stacking. It is also desirable to minimize or avoid repeated use of a single promoter that may cause transcriptional gene silencing. Here we describe a strategy to make polar promoters bidirectional so that one promoter can direct the expression of two genes, one on each end of the promoter. PMID- 11433286 TI - Patenting expressed sequence tags and single nucleotide polymorphisms. PMID- 11433290 TI - Pharmacogenetics initiative galvanizes public and private sectors. PMID- 11433291 TI - Phox domain interaction with PtdIns(3)P targets the Vam7 t-SNARE to vacuole membranes. AB - Specific recognition of phosphoinositides is crucial for protein sorting and membrane trafficking. Protein transport to the yeast vacuole depends on the Vam7 t-SNARE and its phox homology (PX) domain. Here, we show that the PX domain of Vam7 targets to vacuoles in vivo in a manner dependent on phosphatidylinositol 3 phosphate generation. A novel phosphatidylinositol-3-phosphate-binding motif and an exposed loop that interacts with the lipid bilayer are identified by nuclear magnetic resonance spectroscopy. Conservation of key structural and binding site residues across the diverse PX family indicates a shared fold and phosphoinositide recognition function. PMID- 11433293 TI - Mec1p regulates Pds1p levels in S phase: complex coordination of DNA replication and mitosis. AB - Genetic evidence suggests that the securin Pds1p is the target of a late-S-phase checkpoint control. Here we show that Pds1p becomes essential once two-thirds of the genome has been replicated and that the coupling of the completion of genome replication with mitosis relies on the regulation of Pds1p levels. Mec1p is needed to maintain Pds1p levels under S-phase checkpoint conditions. In contrast, Rad53p and Chk1p, needed for the stabilization of Pds1p in the context of the G2 DNA-damage checkpoint pathway, are dispensable. Thus, the Pds1p-dependent late-S phase checkpoint pathway couples replication with mitosis but is mechanistically distinct from the G2 DNA-damage checkpoint. Finally, we show that the inhibition of spindle elongation in early S phase, controlled by the Mec1p/Rad53p branch, is not regulated via Pds1p/Esp1p. This can mechanistically explain the need for branched S-phase checkpoint controls. PMID- 11433294 TI - PAR-1 is a Dishevelled-associated kinase and a positive regulator of Wnt signalling. AB - Wnt signalling regulates beta-catenin-dependent developmental processes through the Dishevelled protein (Dsh). Dsh regulates two distinct pathways, one mediated by beta-catenin and the other by Jun kinase (JNK). We have purified a Dsh associated kinase from Drosophila that encodes a homologue of Caenorhabditis elegans PAR-1, a known determinant of polarity during asymmetric cell divisions. Treating cells with Wnt increases endogenous PAR-1 activity coincident with Dsh phosphorylation. PAR-1 potentiates Wnt activation of the beta-catenin pathway but blocks the JNK pathway. Suppressing endogenous PAR-1 function inhibits Wnt signalling through beta-catenin in mammalian cells, and Xenopus and Drosophila embryos. PAR-1 seems to be a positive regulator of the beta-catenin pathway and an inhibitor of the JNK pathway. These findings show that PAR-1, a regulator of polarity, is also a modulator of Wnt-beta-catenin signalling, indicating a link between two important developmental pathways. PMID- 11433295 TI - Msps protein is localized to acentrosomal poles to ensure bipolarity of Drosophila meiotic spindles. AB - The female meiotic spindle is commonly formed in a centrosome-independent manner. Here we report the identification of proteins at acentrosomal poles in the female meiotic spindle of Drosophila. The acentrosomal poles contain at least two proteins, Mini-spindles (Msps) and D-TACC, which are also associated with mitotic centrosomes. These proteins interact with one another and are both required for maintaining the bipolarity of acentrosomal spindles. The polar localization of Msps is dependent on D-TACC and Ncd, a kinesin-like microtubule motor. We propose that the polar localization of Msps mediated by D-TACC and Ncd may be crucial for the stabilization of meiotic spindle bipolarity. PMID- 11433296 TI - Msps/XMAP215 interacts with the centrosomal protein D-TACC to regulate microtubule behaviour. AB - The XMAP215/ch-TOG/Msps family of microtubule-associated proteins (MAPs) promote microtubule growth in vitro and are concentrated at centrosomes in vivo. We show here that Msps (mini-spindles protein) interacts with the centrosomal protein D TACC, and that this interaction strongly influences microtubule behaviour in Drosophila embryos. If D-TACC levels are reduced, Msps does not concentrate at the centrosomes efficiently and the centrosomal microtubules appear to be destabilized. If D-TACC levels are increased, both D-TACC and Msps accumulate around the centrosomes/spindle poles, and the centrosomal microtubules appear to be stabilized. We show that the interaction between D-TACC and Msps is evolutionarily conserved. We propose that D-TACC and Msps normally cooperate to stabilize centrosomal microtubules by binding to their minus ends and binding to their plus ends as they grow out from the centrosome. PMID- 11433297 TI - N-CAM modulates tumour-cell adhesion to matrix by inducing FGF-receptor signalling. AB - Loss of expression of neural cell-adhesion molecule (N-CAM) is implicated in the progression of tumour metastasis. Here we show that N-CAM modulates neurite outgrowth and matrix adhesion of beta-cells from pancreatic tumours by assembling a fibroblast-growth-factor receptor-4 (FGFR-4) signalling complex, which consists of N-cadherin, FGFR-4, phospholipase C gamma (PLC-gamma), the adaptor protein FRS2, pp60(c-src), cortactin and growth-associated protein-43 (GAP-43). Dominant negative FGFR-4, inhibitors of FGFR signalling and anti-beta(1)-integrin antibodies repress matrix adhesion induced by N-CAM. FGF ligands can replace N CAM in promoting matrix adhesion but not neurite outgrowth. The results indicate that N-CAM stimulates beta1-integrin-mediated cell-matrix adhesion by activating FGFR signalling. This is a potential mechanism for preventing the dissemination of metastatic tumour cells. PMID- 11433298 TI - SNX3 regulates endosomal function through its PX-domain-mediated interaction with PtdIns(3)P. AB - The sorting nexin (SNX) protein family is implicated in regulating membrane traffic, but the mechanism is still unknown. We show that SNX3 is associated with the early endosome through a novel motif (PX domain) capable of interaction with phosphatidylinositol-3-phosphate (PtdIns(3)P). Overexpression of SNX3 alters endosomal morphology and delays transport to the lysosome. Transport from the early to the recycling endosome is affected upon microinjection of SNX3 antibodies. Our results highlight a novel mechanism by which SNX proteins regulate traffic and uncover a novel class of effectors for PtdIns(3)P. PMID- 11433299 TI - Acetylation control of the retinoblastoma tumour-suppressor protein. AB - The retinoblastoma tumour-suppressor protein (pRb) and p300/CBP co-activator proteins are important for control of proliferation and in tumour cells these are sequestered by viral oncoproteins such as E1A. pRb is involved in negatively regulating growth, and p300/CBP proteins have histone acetyltransferase (HAT) activity, which influences gene expression. Although it is known that phosphorylation by G1 cyclin-dependent kinases (CDKs) regulates pRb activity, the nature and role of other post-translational modifications is not understood. Here we identify acetylation as a new type of modification and level of control in pRb function. Adenovirus E1A, which binds p300/CBP through an amino-terminal transformation-sensitive domain, stimulates the acetylation of pRb by recruiting p300 and pRb into a multimeric-protein complex. Furthermore, pRb acetylation is under cell-cycle control, and acetylation hinders the phosphorylation of pRb by cyclin-dependent kinases. pRb binds more strongly when acetylated to the MDM2 oncoprotein, which indicates that acetylation may regulate protein-protein interactions in the pRb pathway. The acetylation of pRb defines a new level of cell-cycle control mediated by HAT. Furthermore, our results establish a relationship between p300, pRb and acetylation in which E1A acts to recruit and target a cellular HAT activity to pRb. PMID- 11433300 TI - The PX domains of p47phox and p40phox bind to lipid products of PI(3)K. AB - PX domains are found in a variety of proteins that associate with cell membranes, but their molecular function has remained obscure. We show here that the PX domains in p47phox and p40phox subunits of the phagocyte NADPH oxidase bind to phosphatidylinositol-3,4-bisphosphate (PtdIns(3,4)P(2)) and phosphatidylinositol 3-phosphate (PtdIns(3)P), respectively. We also show that an Arg-to-Gln mutation in the PX domain of p47phox, which is found in patients with chronic granulomatous disease, eliminates phosphoinositide binding, as does the analogous mutation in the PX domain of p40phox. The PX domain of p40phox localizes specifically to PtdIns(3)P-enriched early endosomes, and this localization is disrupted by inhibition of phosphoinositide-3-OH kinase (PI(3)K) or by the Arg-to Gln point mutation. These findings provide a molecular foundation to understand the role of PI(3)K in regulating neutrophil function and inflammation, and to identify PX domains as specific phosphoinositide-binding modules involved in signal transduction events in eukaryotic cells. PMID- 11433301 TI - PtdIns(3)P regulates the neutrophil oxidase complex by binding to the PX domain of p40(phox). AB - The production of reactive oxygen species (ROS) by neutrophils has a vital role in defence against a range of infectious agents, and is driven by the assembly of a multi-protein complex containing a minimal core of five proteins: the two membrane-bound subunits of cytochrome b(558) (gp91(phox) and p22(phox)) and three soluble factors (GTP-Rac, p47(phox) and p67(phox) (refs 1, 2). This minimal complex can reconstitute ROS formation in vitro in the presence of non physiological amphiphiles such as SDS. p40(phox) has subsequently been discovered as a binding partner for p67(phox) (ref. 3), but its role in ROS formation is unclear. Phosphoinositide-3-OH kinases (PI(3)Ks) have been implicated in the intracellular signalling pathways coordinating ROS formation but through an unknown mechanism. We show that the addition of p40(phox) to the minimal core complex allows a lipid product of PI(3)Ks, phosphatidylinositol 3-phosphate (PtdIns(3)P), to stimulate specifically the formation of ROS. This effect was mediated by binding of PtdIns(3)P to the PX domain of p40(phox). These results offer new insights into the roles for PI(3)Ks and p40(phox) in ROS formation and define a cellular ligand for the orphan PX domain. PMID- 11433302 TI - Novel mechanism of Wnt signalling inhibition mediated by Dickkopf-1 interaction with LRP6/Arrow. AB - Wnt signalling has an important role in cell fate determination, tissue patterning and tumorigenesis. Secreted antagonists of Wnt include Frizzled (Fz) related proteins (FRPs), Cerberus, Wnt inhibitory factor (WIF) and Dickkopf (Dkk). FRPs, Cerberus and WIF have all been shown to act by binding and sequestering Wnt. We report a novel mechanism of Wnt-signalling inhibition by human Dkk-1. Dkk-1 demonstrated no interaction with Wnt but bound a single cell surface site with high affinity (K(D) = 0.39 nM). Its receptor was detectable in a complex with a relative molecular mass of 240,000 (M(r) 240K) with [(125)I] Dkk 1 by covalent affinity cross-linking. Wnt signalling through beta-catenin is mediated by the Fz receptor and a recently identified low-density-lipoprotein receptor-related co-receptor, LRP6/Arrow. Overproduction of the 200K LRP6 protein, but not of Fz, strikingly increased Dkk-1 binding as well as the amount of the 240K cross-linked complex, which was shown to be composed of Dkk-1 and LRP6. Moreover, Dkk-1 function was completely independent of Fz but LRP6 dramatically interfered with the Dkk-1 inhibition of Wnt signalling. Thus, unlike Wnt antagonists, which exert their effects by molecular mimicry of Fz or Wnt sequestration through other mechanisms, Dkk-1 specifically inhibits canonical Wnt signalling by binding to the LRP6 component of the receptor complex. PMID- 11433303 TI - Yeast Eps15-like endocytic protein, Pan1p, activates the Arp2/3 complex. AB - Longstanding evidence supports a role for actin in endocytosis; an intact actin cytoskeleton is required for endocytosis in yeast, and drugs that inhibit actin polymerization inhibit endocytosis in both yeast and mammalian cells. The yeast Arp2/3 complex is required for the internalization step of endocytosis. In addition, some early endocytic events in mammalian cells are associated with the formation of actin tails similar to those generated by activated Arp2/3 complex. However, until now no Arp2/3 complex activator has been identified among proteins known to mediate early steps in endocytosis. Here we show that the yeast endocytic protein Pan1p binds to and activates the Arp2/3 complex. Genetic interactions between PAN1 and mutants of Arp2/3 subunits, or of the Arp2/3 activator LAS17, provide evidence for this activity in vivo. We suggest that Pan1p forms the core of an endocytic complex and physically couples actin polymerization nucleated by the Arp2/3 complex to the endocytic machinery, thus providing the forces necessary for endocytosis. PMID- 11433305 TI - Morphogens: how big is the big picture? AB - Morphogens are in the front line just now. Here I trace how the concept of a morphogen has evolved over the past 100 years and step a little beyond what we already know. PMID- 11433306 TI - Straight or split: signals to transcription. AB - Since the discovery of substance in serum media that are able to drive cells into proliferation and/or differentiation, investigators have tried to understand how such signalling molecules can influence cells to change their behaviour. The complex nature of the responses to signals, and the equally complex signalling pathway leading to those responses, have made life difficult for the researcher. However, recent evidence obtained in genetic developmental systems indicates that a multiplicity of downstream events can be accomplished by regulation of the activity of just one transcription factor. PMID- 11433307 TI - The fatal detachment. PMID- 11433308 TI - PAR-1 helps Wnt to get rid of JNK. PMID- 11433309 TI - TACCing down the spindle poles. PMID- 11433310 TI - Converging on extension. PMID- 11433311 TI - Pathway specificity for Met signalling. PMID- 11433314 TI - Powerful ideas driven by simple tools: lessons from experimental embryology. AB - In developmental biology, as in all scientific fields, conceptual advances are tightly coupled to technological innovation. In this review, we trace the evolution of techniques in experimental embryology, from classical ablation to the latest methods utilizing in vivo electroporation, with lens induction as a linking theme. PMID- 11433316 TI - Much ado about data. PMID- 11433315 TI - 'Super glue'. AB - Cell--cell adhesion is a significant mechanical component of cell and tissue structure. However, cell contacts are not just static mechanical structures: they are integrated into the cytoskeletal and signalling processes of the cell. The formation and remodelling of cell contacts are basic to both tissue morphogenesis and, after damage, wound repair. Loss of adhesion accompanies tumour metastasis. The interplay between these processes was a major theme of a recent joint meeting of the British Societies of Cell Biology and Developmental Biology on 'Cell and Tissue Morphogenesis' in Brighton, UK (3--6 April 2001). PMID- 11433317 TI - A Campylobacter jejuni gene associated with immune-mediated neuropathy. PMID- 11433318 TI - Animal cloning experiments still banned in Italy. PMID- 11433319 TI - The death of just another AIDS orphan? PMID- 11433320 TI - O'Toole is chief of UK cancer research. PMID- 11433321 TI - US plans large scale environmental study. PMID- 11433323 TI - Gene therapy researchers bemoan dual regulations. PMID- 11433324 TI - New addiction research unlikely to be applied. PMID- 11433325 TI - Gates Foundation hires CDC AIDS boss. PMID- 11433326 TI - . . . and funds meningitis vaccine work. PMID- 11433328 TI - Shigekazu Nagata. AB - Having originally researched the activities of the potent hematopeotic stimulator of bone-marrow cells, granulocyte colony stimulating factor, Shigekazu Nagata is better known for his work on apoptosis. Here, one of Japan's most renowned biomedical scientists outlines the path that has taken him full circle: from stimulating cells to grow, to finding out how they die, to tying the two processes together. (Interview by David Cyranoski.) PMID- 11433329 TI - Dendritic cells: On the move from bench to bedside. AB - As dendritic cells increasingly become the adjuvant of choice in new approaches to cancer immunotherapy, a degree of protocol standardization is required to aid future large-scale clinical trials. PMID- 11433332 TI - Minor antigen solves major problem. PMID- 11433333 TI - Uncoupling protein 2: Adiposity angel and diabetes devil? PMID- 11433334 TI - Sensory neurons are PARtial to pain. PMID- 11433336 TI - Life after death: Controlling autoreactive CD8+ T-cell responses. PMID- 11433337 TI - Clearing the smoke on nicotine and angiogenesis. PMID- 11433338 TI - CD4+ T cells: The great escape. PMID- 11433339 TI - Protein-based PCR for prion diseases? PMID- 11433341 TI - Replication-selective virotherapy for cancer: Biological principles, risk management and future directions. AB - In the search for novel cancer therapies that can be used in conjunction with existing treatments, one promising area of research is the use of viral vectors and whole viruses. This review describes the underlying biological principles and current status of the field, outlines approaches for improving clinical effectiveness and discusses the unique safety and regulatory issues surrounding viral therapies. PMID- 11433342 TI - Adoptive transfer of minor histocompatibility antigen-specific T lymphocytes eradicates leukemia cells without causing graft-versus-host disease. AB - Adoptive transfer of T cells reactive to minor histocompatibility antigens has the unmatched ability to eradicate malignant hematopoietic cells. Unfortunately, its use is hampered by the associated graft-versus-host disease. The critical issue of a possible dissociation of the antileukemic effect and graft-versus-host disease by targeting specific minor histocompatibility antigens remains unresolved because of the unknown nature and number of minor histocompatibility antigens necessary or sufficient to elicit anti-leukemic activity and graft versus-host disease. We found that injection of T lymphocytes primed against a single major histocompatibility complex class I-restricted immunodominant minor histocompatibility antigen (B6dom1) caused no graft-versus-host disease but produced a curative anti-leukemic response. Avoidance of graft-versus-host disease required that no other host-reactive T cells be co-injected with T cells primed with B6dom1. Here we show that effective and non-toxic immunotherapy of hematologic malignancies can be achieved by targeting a single immunodominant minor histocompatibility antigen. PMID- 11433343 TI - CD4+ T-cell-epitope escape mutant virus selected in vivo. AB - Mutations in viral genomes that affect T-cell-receptor recognition by CD8+ cytotoxic T lymphocytes have been shown to allow viral evasion from immune surveillance during persistent viral infections. Although CD4+ T-helper cells are crucially involved in the maintenance of effective cytotoxic T-lymphocyte and neutralizing-antibody responses, their role in viral clearance and therefore in imposing similar selective pressures on the virus is unclear. We show here that transgenic virus-specific CD4+ Tcells, transferred into mice persistently infected with lymphocytic choriomeningitis virus, select for T-helper epitope mutant viruses that are not recognized. Together with the observed antigenic variation of the same T-helper epitope during polyclonal CD4+ T-cell responses in infected pore-forming protein-deficient C57BL/6 mice, this finding indicates that viral escape from CD4+ T lymphocytes is a possible mechanism of virus persistence. PMID- 11433344 TI - Epithelial secretion of C3 promotes colonization of the upper urinary tract by Escherichia coli. AB - To assess the role of complement in renal infection, we studied a model of Escherichia coli-induced pyelonephritis in mice deficient in complement components C3 and C4. Renal infection occurred less frequently in C3- and C4 deficient mice compared with wild-type mice. In vitro, renal epithelial cells internalized fewer bacteria in the absence of C3 or in the presence of blockade of C3 bound to the bacteria. Moreover, upregulation of epithelial C3 production by stimulation with lipopolysaccharide enhanced bacterial internalization. Here we provide evidence that uropathogenic E. coli might use host C3 to invade the renal epithelium and that local complement production is sufficient for the bacteria to achieve this effect. PMID- 11433345 TI - Apoptotic cells overexpress vinculin and induce vinculin-specific cytotoxic T cell cross-priming. AB - Here we show that apoptotic cells overexpress vinculin and are ingested by dendritic cells, which subsequently cross-prime vinculin-specific cytotoxic T lymphocytes (CTLs). Successful cross-priming requires that the apoptotic cells provide maturation signals to dendritic cells through CD40-CD40 ligand (CD40L) interactions. If apoptotic cells are CD40L-, the help of a third T cell is needed for priming, indicating a regulatory role for apoptotic cells in determining priming or tolerance. Vinculin-specific CTL priming is also related to apoptosis in vivo, given that in HIV-seropositive individuals, the frequency of specific CTLs depends on the proportion of peripheral CD40L+ apoptotic cells. PMID- 11433346 TI - Fanconi anemia group C protein prevents apoptosis in hematopoietic cells through redox regulation of GSTP1. AB - The Fanconi anemia group C protein (FANCC) plays an important role in hematopoiesis by ensuring the survival of hematopoietic progenitor cells through an unknown mechanism. We investigated the function of FANCC by identifying FANCC binding proteins in hematopoietic cells. Here we show that glutathione S transferase P1-1 (GSTP1) interacts with FANCC, and that overexpression of both proteins in a myeloid progenitor cell line prevents apoptosis following factor deprivation. FANCC increases GSTP1 activity after the induction of apoptosis. GSTP1 is an enzyme that catalyzes the detoxification of xenobiotics and by products of oxidative stress, and it is frequently upregulated in neoplastic cells. Although FANCC lacks homology with conventional disulfide reductases, it functions by preventing the formation of inactivating disulfide bonds within GSTP1 during apoptosis. The prevention of protein oxidation by FANCC reveals a novel mechanism of enzyme regulation during apoptosis and has implications for the treatment of degenerative diseases with thiol reducing agents. PMID- 11433347 TI - Proteinase-activated receptor-2 and hyperalgesia: A novel pain pathway. AB - Using a combined pharmacological and gene-deletion approach, we have delineated a novel mechanism of neurokinin-1 (NK-1) receptor-dependent hyperalgesia induced by proteinase-activated receptor-2 (PAR2), a G-protein-coupled receptor expressed on nociceptive primary afferent neurons. Injections into the paw of sub-inflammatory doses of PAR2 agonists in rats and mice induced a prolonged thermal and mechanical hyperalgesia and elevated spinal Fos protein expression. This hyperalgesia was markedly diminished or absent in mice lacking the NK-1 receptor, preprotachykinin-A or PAR2 genes, or in rats treated with a centrally acting cyclooxygenase inhibitor or treated by spinal cord injection of NK-1 antagonists. Here we identify a previously unrecognized nociceptive pathway with important therapeutic implications, and our results point to a direct role for proteinases and their receptors in pain transmission. PMID- 11433348 TI - Endocannabinoids acting at vascular CB1 receptors mediate the vasodilated state in advanced liver cirrhosis. AB - Advanced cirrhosis is associated with generalized vasodilation of unknown origin, which contributes to mortality. Cirrhotic patients are endotoxemic, and activation of vascular cannabinoid CB1 receptors has been implicated in endotoxin induced hypotension. Here we show that rats with biliary cirrhosis have low blood pressure, which is elevated by the CB1 receptor antagonist SR141716A. The low blood pressure of rats with CCl4-induced cirrhosis was similarly reversed by SR141716A, which also reduced the elevated mesenteric blood flow and portal pressure. Monocytes from cirrhotic but not control patients or rats elicited SR141716A-sensitive hypotension in normal recipient rats and showed significantly elevated levels of anandamide. Compared with non-cirrhotic controls, in cirrhotic human livers there was a three-fold increase in CB1 receptors on isolated vascular endothelial cells. These results implicate anandamide and vascular CB1 receptors in the vasodilated state in advanced cirrhosis and indicate a novel approach for its management. PMID- 11433349 TI - Nicotine stimulates angiogenesis and promotes tumor growth and atherosclerosis. AB - We provide anatomic and functional evidence that nicotine induces angiogenesis. We also show that nicotine accelerates the growth of tumor and atheroma in association with increased neovascularization. Nicotine increased endothelial cell growth and tube formation in vitro, and accelerated fibrovascular growth in vivo. In a mouse model of hind-limb ischemia, nicotine increased capillary and collateral growth, and enhanced tissue perfusion. In mouse models of lung cancer and atherosclerosis, we found that nicotine enhanced lesion growth in association with an increase in lesion vascularity. These effects of nicotine were mediated through nicotinic acetylcholine receptors at nicotine concentrations that are pathophysiologically relevant. The endothelial production of nitric oxide, prostacyclin and vascular endothelial growth factor might have a role in these effects. PMID- 11433350 TI - A link between diabetes and atherosclerosis: Glucose regulates expression of CD36 at the level of translation. AB - Both the risk and the rate of development of atherosclerosis are increased in diabetics, but the mechanisms involved are unknown. Here we report a glucose mediated increase in CD36 mRNA translation efficiency that results in increased expression of the macrophage scavenger receptor CD36. Expression of CD36 was increased in endarterectomy lesions from patients with a history of hyperglycemia. Macrophages that were differentiated from human peripheral blood monocytes in the presence of high glucose concentrations showed increased expression of cell-surface CD36 secondary to an increase in translational efficiency of CD36 mRNA. We obtained similar data from primary cells isolated from human vascular lesions, and we found that glucose sensitivity is a function of ribosomal reinitiation following translation of an upstream open reading frame (uORF). Increased translation of macrophage CD36 transcript under high glucose conditions provides a mechanism for accelerated atherosclerosis in diabetics. PMID- 11433351 TI - Apolipoprotein B secretion and atherosclerosis are decreased in mice with phospholipid-transfer protein deficiency. AB - Increased secretion and levels of ApoB-containing lipoproteins (BLp) commonly occur in familial hyperlipidemia, obesity and diabetes. The plasma phospholipid transfer protein (PLTP) is known to mediate transfer of phospholipids between BLp and HDL during their intravascular metabolism. To address a possible role of PLTP in dyslipidemia and atherogenesis, we bred mice deficient in the gene encoding PLTP (PLTP-deficient mice) using different hyperlipidemic mouse strains. In ApoB transgenic and ApoE-deficient backgrounds, PLTP deficiency resulted in reduced production and levels of BLp and markedly decreased atherosclerosis. BLp secretion was diminished in hepatocytes from ApoB-transgenic PLTP-deficient mice, a defect that was corrected when PLTP was reintroduced in adenovirus. The studies reveal a major, unexpected role of PLTP in regulating the secretion of BLp and identify PLTP as a therapeutic target. PMID- 11433352 TI - High-density lipoprotein binding to scavenger receptor-BI activates endothelial nitric oxide synthase. AB - Atherosclerosis is the primary cause of cardiovascular disease, and the risk for atherosclerosis is inversely proportional to circulating levels of high-density lipoprotein (HDL) cholesterol. However, the mechanisms by which HDL is atheroprotective are complex and not well understood. Here we show that HDL stimulates endothelial nitric oxide synthase (eNOS) in cultured endothelial cells. In contrast, eNOS is not activated by purified forms of the major HDL apolipoproteins ApoA-I and ApoA-II or by low-density lipoprotein. Heterologous expression experiments in Chinese hamster ovary cells reveal that scavenger receptor-BI (SR-BI) mediates the effects of HDL on the enzyme. HDL activation of eNOS is demonstrable in isolated endothelial-cell caveolae where SR-BI and eNOS are colocalized, and the response in isolated plasma membranes is blocked by antibodies to ApoA-I and SR-BI, but not by antibody to ApoA-II. HDL also enhances endothelium- and nitric-oxide-dependent relaxation in aortae from wild-type mice, but not in aortae from homozygous null SR-BI knockout mice. Thus, HDL activates eNOS via SR-BI through a process that requires ApoA-I binding. The resulting increase in nitric-oxide production might be critical to the atheroprotective properties of HDL and ApoA-I. PMID- 11433353 TI - Positron emission tomography imaging for herpes virus infection: Implications for oncolytic viral treatments of cancer. AB - Molecular therapy using viruses would benefit greatly from a non-invasive modality for assessing dissemination of viruses. Here we investigated whether positron emission tomography (PET) scanning using [(124)I]-5-iodo-2'-fluoro-1 beta-d-arabinofuranosyl-uracil (FIAU) could image cells infected with herpes simplex viruses (HSV). Using replication-competent HSV-1 oncolytic viruses with thymidine kinase (TK) under control of different promoters, we demonstrate that viral infection, proliferation and promoter characteristics all interact to influence FIAU accumulation and imaging. In vivo, as few as 1 x 107 viral particles injected into a 0.5-cm human colorectal tumor can be detected by [(124)I]FIAU PET imaging. PET signal intensity is significantly greater at 48 hours compared with that at 8 hours after viral injection, demonstrating that PET scanning can detect changes in TK activity resulting from local viral proliferation. We also show the ability of FIAU-PET scanning to detect differences in viral infectivity at 0.5 log increments. Non-invasive imaging might be useful in assessing biologically relevant distribution of virus in therapies using replication-competent HSV. PMID- 11433354 TI - In vivo measurement of gene expression, angiogenesis and physiological function in tumors using multiphoton laser scanning microscopy. AB - Intravital microscopy coupled with chronic animal window models has provided stunning insight into tumor pathophysiology, including gene expression, angiogenesis, cell adhesion and migration, vascular, interstitial and lymphatic transport, metabolic microenvironment and drug delivery. However, the findings to date have been limited to the tumor surface (< 150 microm). Here, we show that the multiphoton laser-scanning microscope can provide high three-dimensional resolution of gene expression and function in deeper regions of tumors. These insights could be critical to the development of novel therapeutics that target not only the tumor surface, but also internal regions. PMID- 11433356 TI - Genome annotation: from sequence to biology. AB - The genome sequence of an organism is an information resource unlike any that biologists have previously had access to. But the value of the genome is only as good as its annotation. It is the annotation that bridges the gap from the sequence to the biology of the organism. The aim of high-quality annotation is to identify the key features of the genome - in particular, the genes and their products. The tools and resources for annotation are developing rapidly, and the scientific community is becoming increasingly reliant on this information for all aspects of biological research. PMID- 11433357 TI - Evolving responsively: adaptive mutation. AB - A basic principle of genetics is that the likelihood that a particular mutation occurs is independent of its phenotypic consequences. The concept of adaptive mutation seemed to challenge this principle with the discoveries of mutations stimulated by stress, some of which allow adaptation to the stress. The emerging mechanisms of adaptive genetic change cast evolution, development and heredity into a new perspective, indicating new models for the genetic changes that fuel these processes. PMID- 11433358 TI - The arms race is ancient history in Arabidopsis, the wildflower. AB - Plant pathology was born after the nineteenth-century potato famine, and since then insightful genetic experiments have contributed to the great progress in our understanding of disease control. Our current view of plant resistance focuses on numerous polymorphic resistance loci, which contain genes known as R genes. The complete sequence of the Arabidopsis thaliana genome provides a framework for exploring the 'big bang' of R genes that occurred and how R genes evolved in plants from their associations with microorganisms, and for improving strategies for more sustainable deployment of disease resistance in crops. PMID- 11433359 TI - Functional genomics and the study of development, variation and evolution. AB - It has rarely been possible to connect the developmental and evolutionary branches of genetics, particularly with regard to the precise changes in the molecular control of development that are responsible for phenotypic variation and evolution. Making such connections will require a high-resolution molecular description of the genetic networks that underlie development and an understanding of their responses to genetic and environmental variation. Functional genomics approaches to development and evolution promise to accelerate the research necessary to accomplish these goals. PMID- 11433360 TI - Placental development: lessons from mouse mutants. AB - The placenta is the first organ to form during mammalian embryogenesis. Problems in its formation and function underlie many aspects of early pregnancy loss and pregnancy complications in humans. Because the placenta is critical for survival, it is very sensitive to genetic disruption, as reflected by the ever-increasing list of targeted mouse mutations that cause placental defects. Recent studies of mouse mutants with disrupted placental development indicate that signalling interactions between the placental trophoblast and embryonic cells have a key role in placental morphogenesis. Furthering our understanding of mouse trophoblast development should provide novel insights into human placental function. PMID- 11433361 TI - The evolution of isochores. AB - One of the most striking features of mammalian chromosomes is the variation in G+C content that occurs over scales of hundreds of kilobases to megabases, the so called 'isochore' structure of the human genome. This variation in base composition affects both coding and non-coding sequences and seems to reflect a fundamental level of genome organization. However, although we have known about isochores for over 25 years, we still have a poor understanding of why they exist. In this article, we review the current evidence for the three main hypotheses. PMID- 11433362 TI - Molecular metaphors: the gene in popular discourse. AB - Geneticists deploy a striking range of metaphors to communicate their science, to promote its value and to suggest its social meaning to the public. So too, critics of science and special interest groups use metaphorical constructs to express their concerns about the implications of the 'genetic revolution'. Through metaphors, genetics can seem a source of salvation or a means of exploitation, a boon to health or a source of risk. This paper is a critical review of the metaphors used to communicate genetic information to the public. PMID- 11433363 TI - Linking class-switch recombination with somatic hypermutation. AB - The recent discovery of a molecular link between two apparently different genetic alteration events--class-switch recombination and somatic hypermutation--has led to the idea that the recognition and cleavage of target DNA in these two events might be mediated by similar or identical molecules to those involved in RNA editing. This could mean that the complexity of mammalian genetic information may be enriched by an interplay between RNA editing and DNA modification. PMID- 11433364 TI - How proteins move lipids and lipids move proteins. AB - Cells determine the bilayer characteristics of different membranes by tightly controlling their lipid composition. Local changes in the physical properties of bilayers, in turn, allow membrane deformation, and facilitate vesicle budding and fusion. Moreover, specific lipids at specific locations recruit cytosolic proteins involved in structural functions or signal transduction. We describe here how the distribution of lipids is directed by proteins, and, conversely, how lipids influence the distribution and function of proteins. PMID- 11433365 TI - The function and synthesis of ribosomes. AB - Structural analyses of the large and small ribosomal subunits have allowed us to think about how they work in more detail than ever before. The mechanisms that underlie ribosomal synthesis, translocation and catalysis are now being unravelled, with practical implications for the design of antibiotics. PMID- 11433366 TI - Translational control by CPEB: a means to the end. AB - The regulated translation of messenger RNA is essential for cell-cycle progression, establishment of the body plan during early development, and modulation of key activities in the central nervous system. Cytoplasmic polyadenylation, which is one mechanism of controlling translation, is driven by CPEB--a highly conserved, sequence-specific RNA-binding protein that binds to the cytoplasmic polyadenylation element, and modulates translational repression and mRNA localization. What are the features and functions of this multifaceted protein? PMID- 11433367 TI - The bacterial toxin toolkit. AB - Pathogenic bacteria and higher eukaryotes have spent a long time together, leading to a precise understanding of one another's way of functioning. Through rapid evolution, bacteria have engineered increasingly sophisticated weapons to hit exactly where it hurts, interfering with fundamental host functions. However, toxins are not only useful to the bacteria - they have also become an essential asset for life scientists, who can now use them as toolkits to explore cellular processes. PMID- 11433368 TI - DNA transport in bacteria. AB - DNA transport is important in various biological contexts--particularly chromosome segregation and intercellular gene transfer. Recently, progress has been made in understanding the function of a family of bacterial proteins involved in DNA transfer, and we focus here on one of the best-understood members, SpoIIIE. Studies of SpoIIIE-like proteins show that they might couple DNA transport to processes such as cell division, conjugation (mating) and the resolution of chromosome dimers. PMID- 11433369 TI - Programmed cell death and apoptosis: origins of the theory. AB - Interest in the study of apoptosis grew with the recognition that it is a highly regulated process. Such a change in attitude allowed the intellectual and technical breakthroughs that led to the explosive development of this subject. PMID- 11433370 TI - XIAP, the guardian angel. AB - Controlling the activity of caspases is essential for the appropriate execution of cell death and the regulation of cell survival. One cellular inhibitor of apoptosis, XIAP, has emerged as a crucial regulator of caspases, and is itself subject to complex negative regulation. PMID- 11433371 TI - Roles of serine/threonine phosphatases in hippocampal synaptic plasticity. PMID- 11433372 TI - Functional brain development in humans. PMID- 11433373 TI - Genetic regulation of cerebellar development. PMID- 11433374 TI - Alpha-synuclein and neurodegenerative diseases. PMID- 11433375 TI - Metalloproteinases in biology and pathology of the nervous system. PMID- 11433376 TI - Stem cell plasticity--building the brain of our dreams. PMID- 11433377 TI - Hypothalamic integration of central and peripheral clocks. PMID- 11433378 TI - Immunocompetent astrocytes and microglia display major differences in the processing of the invariant chain and in the expression of active cathepsin L and cathepsin S. AB - The role of astrocytes and microglia as antigen-presenting cells in the brain is still controversial. In this study we have analyzed and compared aspects of the molecular machinery that underlies MHC class II trafficking in immunocompetent astrocytes and microglia. We show that IFN-gamma-stimulated microglia possess active cathepsin L and cathepsin S, and efficiently degrade the invariant chain, unlike IFN-gamma-stimulated astrocytes that express cathepsin L but not cathepsin S. The lack of cathepsin S proves to be dramatic for the antigen-presentation capacity of astrocytes, which is nearly abolished when these cells are stimulated by a combination of IFN-gamma and TNF-alpha. TNF-alpha indeed decreases cathepsin L activity as we show here, leading to alterations in invariant chain processing, and hence in MHC class II trafficking in astrocytes. Cystatin C inhibits cathepsin L activity in astrocytes, but does not regulate cathepsin L and cathepsin S activity in microglia. We therefore identify cathepsin L and cathepsin S as key components in the regulation of the immune potential of astrocytes and microglia, and provide evidence for a cell-specific regulation exerted by IFN-gamma and TNF-alpha on the expression and activity of cathepsins. PMID- 11433379 TI - Structural and functional dissection of the cytoplasmic domain of the transmembrane adaptor protein SIT (SHP2-interacting transmembrane adaptor protein). AB - SIT (SHP2-interacting transmembrane adaptor protein) is a recently identified transmembrane adaptor protein, which is expressed in lymphocytes. Its structural properties, in particular the presence of five potential tyrosine phosphorylation sites, suggest involvement of SIT in TCR-mediated recruitment of SH2 domain containing intracellular signaling molecules to the plasma membrane. Indeed, it has recently been demonstrated that SIT inducibly interacts with the SH2 containing protein tyrosine phosphatase 2 (SHP2) via an immunoreceptor tyrosine based inhibition motif (ITIM). Moreover, SIT is capable to inhibit TCR-mediated signals proximal of activation of protein kinase C. However, inhibition of T cell activation by SIT occurs independently of SHP2 binding. The present study was performed to further characterize the molecular interaction between SIT and intracellular effector molecules and to identify the protein(s) mediating its inhibitory function. We demonstrate that SIT not only interacts with SHP2 but also with the adaptor protein Grb2 via two consensus YxN motifs. However, mutation of both Grb2-binding sites also does not influence the inhibitory function of SIT. In contrast, mutation of the tyrosine-based signaling motif Y(168) ASV completely abrogates the ability of SIT to inhibit T cell activation. Co-precipitation experiments revealed that the tyrosine kinase p50(csk) could represent the negative regulatory effector molecule that binds to this motif. PMID- 11433380 TI - Complementarity and redundancy of the binding specificity of HLA-DRB1, -DRB3, DRB4 and -DRB5 molecules. AB - The second HLA-DR molecules, which are encoded by loci different from HLA-DRB1 are weakly polymorphic. Predominant alleles such as HLA-DRB3*0101, HLA-DRB4*0101 and HLA-DRB5*0101 are therefore interesting targets to define antigenic peptides with major impact for the entire population. Strikingly, they have been poorly investigated. Thus we have characterized peptides from the major bee venom allergen that bind efficiently to these molecules and compared them to peptides specific for preponderant HLA-DRB1 molecules. Interestingly, DRB5*0101 and DRB1*0701 molecules share four binding peptides and use some identical anchor residues. Similarities are also found between DRB3*0101 and its haplotype associated molecules DRB1*0301 and DRB1*1301. In sharp contrast, DRB4*0101 exhibits a unique binding specificity, which results from particular structural features of its peptide binding site. Ybeta81 seems to alter the amino acid preferences of the P1 pocket, while Rbeta71, Ebeta74, Nbeta26 and Cbeta13 confer to the P4 pocket a unique topology. Our results show that the two HLA-DR molecules expressed in most haplotypes studied here have mostly complementary binding patterns. Only haplotype HLA-DR52 exhibits peptide binding redundancies. Finally our results document functional similarities among HLA-DR molecules and allow us to propose peptide sequences that might be useful for bee venom immunotherapy. PMID- 11433381 TI - Genetic control of collagen-induced arthritis in a cross with NOD and C57BL/10 mice is dependent on gene regions encoding complement factor 5 and FcgammaRIIb and is not associated with loci controlling diabetes. AB - The nonobese diabetic (NOD) mouse spontaneously develops autoimmune-mediated diseases such as diabetes and Sjogren's syndrome. To investigate whether NOD genes also promote autoimmune-mediated arthritis we established a NOD strain with an MHC class II fragment containing the A(q) class II gene predisposing for collagen induced arthritis (NOD.Q). However, this mouse was resistant to arthritis in contrast to other A(q) expressing strains such as B10.Q and DBA/1. To determine the major resistance factor/s, a genetic analysis was performed. (NOD.Q x B10.Q)F1 mice were resistant, whereas 27% of the (NOD.Q x B10.Q)F2 mice developed severe arthritis. Genetic mapping of 353 F2 mice revealed two loci associated with arthritis. One locus was found on chromosome 2 (LOD score 9.8), at the location of the complement factor 5 (C5) gene. The susceptibility allele was from B10.Q, which contains a productive C5 encoding gene in contrast to NOD.Q. The other significant locus was found on chromosome 1 (LOD score 5.6) close to the Fc-gamma receptor IIb gene, where NOD carried the susceptible allele. An interaction between the two loci was observed, indicating that they operate on the same or on interacting pathways. The genetic control of arthritis is unique in comparison to diabetes, since none of these loci have been identified in analysis of diabetes susceptibility. PMID- 11433382 TI - Endogenous ligands of carbohydrate recognition domains of the mannose receptor in murine macrophages, endothelial cells and secretory cells; potential relevance to inflammation and immunity. AB - The macrophage mannose receptor (MR) has an established role in the phagocytosis of a wide range of microbes, and also functions in viral endocytosis, and clearance of a number of endogenous glycoproteins from the circulation. Its broad ligand specificity is mediated by tandemly linked carbohydrate recognition domains (CRDs). Recent studies suggest that binding or internalization of both natural and synthetic ligands of MR CRDs may modulate macrophage (MPhi ) function, for example to increase cidal capacity or cytokine synthesis. To identify endogenous ligands in the normal mouse we used an Fc-fusion protein (CRD4-7Fc) bearing four of the CRDs of MR. CRD4-7Fc recognized endocytic compartments of cultured MPhi, consistent with lysosomal enzymes being major ligands of MR. CRD4-7Fc also recognized MPhi and some endothelial cells in tissues, and intensely labeled secretory cells of the exocrine pancreas, salivary gland and thyroid. Strongly MR-positive interstitial cells were found in close proximity to the ligand-rich secretory cells, suggesting a role for MR in uptake of secretory glycoproteins, including thyroglobulin which was identified as a novel ligand in vitro. Endocytosis of these ligands by MR may have implications for tissue homeostasis and immunity, including antigen presentation, in secretory organs. PMID- 11433383 TI - A critical role for the T cell receptor alpha-chain connecting peptide domain in positive selection of CD1-independent NKT cells. AB - Natural killer T (NKT) cells are a subset of mature alpha beta TCR(+) cells that co-express NK lineage markers. Whereas most NKT cells express a canonical Valpha14/Vbeta8.2 TCR and are selected by CD1d, a minority of NKT cells express a diverse TCR repertoire and develop independently of CD1d. Little is known about the selection requirements of CD1d-independent NKT cells. We show here that NKT cells develop in RAG-deficient mice expressing an MHC class II-restricted transgenic TCR (Valpha2/Vbeta8.1) but only under conditions that lead to negative selection of conventional T cells. Moreover development of NKT cells in these mice is absolutely dependent upon an intact TCR alpha-chain connecting peptide domain, which is required for positive selection of conventional T cells via recruitment of the ERK signaling pathway. Collectively our data demonstrate that NKT cells can develop as a result of high avidity TCR/MHC class II interactions and suggest that common signaling pathways are involved in the positive selection of CD1d-independent NKT cells and conventional T cells. PMID- 11433384 TI - Elevated Bcl-2 is not a causal event in the positive selection of T cells. AB - T cell development is characterized by the induction of apoptosis in most immature thymocytes and the rescue from apoptosis of a small proportion of cells by the process of positive selection.Up-regulation of the anti-apoptotic molecule Bcl-2 is associated with thymocytes undergoing positive selection and a bcl-2 transgene promotes the generation of mature T cells. In contrast,mice transgenic for the pro-apoptotic molecule Bax show impaired T cell maturation. We have used fetal thymic organ culture to determine the action of Bcl-2 and Bax on positive selection of thymocytes. Our data show that Bcl-2 and Bax do not alter the number of thymocytes positively selected by a defined peptide ligand. This implies that Bcl-2 and Bax alter the production of mature T cells in vivo by influencing thymocyte viability rather than by direct action on positive selection. It also presents a solution to the 'chicken-and-egg' scenario relating to Bcl-2 up regulation and positive selection. The data suggest that the up-regulation of Bcl 2 associated with T cell maturation is a consequence of positive selection rather than a cause of it. PMID- 11433385 TI - Role of NFkappaB in antigen presentation and development of regulatory T cells elucidated by treatment of dendritic cells with the proteasome inhibitor PSI. AB - Dendritic cells (DC) are the most potent antigen-presenting cells for naive T cells, due to their high expression of MHC and costimulatory molecules, but relatively little is known about the biochemical pathways that regulate this function. We used the proteasome inhibitor N-benzyloxycarbonyl-Ile-Glu(O-tert butyl)-Ala-leucinal (PSI) to demonstrate that DC antigen presentation is NFkappaB dependent. As PSI is not a specific inhibitor of NFkappaB, we reproduced this finding using a very specific approach, namely adenoviral gene transfer of IkappaBalpha, the naturally occurring inhibitor of NFkappaB. The mechanism for this inhibition of DC antigen presentation involves at least three aspects of antigen presenting function: down-regulation of HLA class II, down-regulation of CD86, and inhibition of the immunostimulatory cytokines IL-12 and TNF-alpha. In the light of the marked down-regulation of antigen-presentation cell function, it was of interest to investigate what effects exposure to PSI-treated DC might have on T cell function. It was found that immunological tolerance was induced, as challenge of T cells previously exposed to PSI-treated DC, with normal DC from the same donor did not restore their response, despite the presence of viable T cells. There were also changes in T cell surface markers, with down-regulation of CD3 and CD25 expression, and inhibition of the production of Th1 cytokines like IL-2 and IFN-gamma. These results demonstrates that NFkappaB is an effective target for blocking DC antigen presentation and inhibiting T cell-dependent immune responses, and this has implications for the development of therapeutic agents for use in multiple conditions, including transplantation, allergy and autoimmune diseases. PMID- 11433386 TI - T cell recognition of rat myelin basic protein as a TCR antagonist inhibits reciprocal activation of antigen-presenting cells and engenders resistance to experimental autoimmune encephalomyelitis. AB - The aim of this study was to assess whether T cell recognition of myelin basic protein (MBP) as a partially antagonistic self antigen regulates the reciprocal activation of professional antigen-presenting cells (APC). This study focused on the rat 3H3 T cell clone that recognized guinea pig (GP) MBP as a full agonist and self rat (R) MBP as a partial agonist. In cultures of 3H3 T cells and splenic APC, the agonist GPMBP elicited several responses by splenic APC, including production of nitric oxide, down-regulation of I-A, induction of B7.1 and B7.2, and prolongation of APC survival. RMBP stimulated a partial increase in production of nitric oxide, partially promoted survival of splenic APC, but did not alter expression of I-A, B7.1, or B7.2 on splenic APC. In the presence ofGPMBP, RMBP antagonized agonist-stimulated induction of B7 molecules, reversed the loss of I-A, and promoted the generation of I-A(+), costimulus-deficient APC. Furthermore, 3H3 T cells cultured with RMBP and irradiated splenocytes reduced the severity of EAE upon adoptive transfer into naive rat recipients subsequently challenged with an encephalitogenic dose of GPMBP/CFA. Overall, this study indicates that T cell receptor antagonism blocks T cell activation, inhibits feedback activation of splenic APC, and promotes T cell-dependent regulatory activities in EAE. PMID- 11433387 TI - Identification of committed NK cell progenitors in adult murine bone marrow. AB - Natural killer (NK) cells play important roles in innate immunity by lysing tumor and virally infected cells and by producing cytokines including interferon-gamma. While NK cell progenitors have been described in the fetal thymus, NK cell generation from hematopoietic stem cells (HSC) in the bone marrow (BM) occurs throughout life, and in athymic mice and humans. Interleukin (IL)-15 promotes NK development in vitro and is essential for the generation of normal numbers of NK cells in vivo. By characterizing BM cells expressing IL-15 receptor components, we found marked heterogeneity within the IL-2 receptor beta chain(+) (CD122(+)) subset, which included cells uniquely committed to the NK lineage. These CD122(+) NK cell precursors (NKP) are negative for markers used to identify mature NK cells, including NK1.1, DX5 and members of Ly-49 family, and fail to demonstrate natural cytotoxicity against susceptible target cells. In vitro culture of NKP generates mature lytic NK1.1(+) cells at high frequencies, while they do not give rise to T, B, myeloid or erythroid cells under appropriate conditions. NKP lack transcripts associated with early B and T cell differentiation (pTalpha, lambda5 and CD3epsilon), but express a group of genes (IL-15Ralpha, Id2, GATA-3 and Ets 1) and the 2B4 marker, which may define NK cell commitment. We propose that NKP represent the earliest adult BM precursor uniquely restricted to the NK cell lineage. PMID- 11433388 TI - A MAGE-1 peptide recognized on HLA-DR15 by CD4(+) T cells. AB - Antigens encoded by MAGE genes and recognized by T cells are of interest for cancer immunotherapy because of their strict tumoral specificity and because they are shared by many tumors. Several MAGE-1 peptide that are recognized by CD8(+) cytolytic T lymphocytes have been used in therapeutic vaccination trials. To obtain anti-tumor immune response, vaccines combining peptides recognized by CD8(+) and peptides recognized by CD4(+) T cells might be optimal. We focused therefore on the identification of MAGE peptides recognized by CD4(+) T cells. We report here the identification of MAGE-1 epitope EYVIKVSARVRF, which is presented to CD4(+) T lymphocytes by HLA-DR15. This HLA allele is present in 29 % of Asians and 17 % of Caucasians. PMID- 11433389 TI - Reevaluation of the origin of CD44(high) "memory phenotype" CD8 T cells: comparison between memory CD8 T cells and thymus-independent CD8 T cells. AB - CD44(high)CD8 T cells in naive mice, which increase with age, and are often referred to as memory CD8 T cells. However, since thymus-independent CD8 T cells have also been shown to be CD44(high), the origin of the CD44(high)CD8 T cells in naive mice remains unclear. In this study, we compared the characteristics of memory CD8 T cells and thymus-independent CD8 T cells in TCR transgenic mice to clarify the origin of the CD44(high)CD8 T cells in naive normal mice. The memory and thymus-independent CD8 T cells showed differences in surface molecules, spontaneous cell death, cytokine production, and response to IL-2R binding of cytokines. Importantly, the "memory phenotype" CD8 T cells in naive normal mice showed similar characteristics to the thymus-independent CD8 T cells, but differed greatly from "true" memory CD8 T cells in the TCR transgenic mice. Therefore, we conclude that a significant part of the CD44(high) memory phenotype CD8 T cells in naive normal mice represents thymus-independent CD8 T cells, which may participate in age-related changes in immune responses. PMID- 11433390 TI - Preferential Th1 profile of T helper cell responses in X-linked (Bruton's) agammaglobulinemia. AB - X-linked agammaglobulinemia (XLA) is a primary immunodeficiency of the B-cell compartment caused by a defective gene encoding for the tyrosine kinase (btk) essential for B cell differentiation. Affected males undergo recurrent pyogenic infections and deficient immunoglobulin production. Peripheral blood T cells from 6 XLA patients and 6 matched healthy controls were stimulated with either PHA or tetanus toxoid (TT) and T cell clones obtained were compared for their cytokine profile. In the series of PHA-induced or TT-specific CD4(+) T cell clones derived from XLA patients, the Th1 profile was predominant (63 and 65 %, respectively). Upon stimulation with TT, the proportion of activated T cells from XLA that expressed the IFN-gamma -associated LAG-3 activation molecule was higher than in control T cells (51 vs. 25 %), whereas the expression of the IL-4-associated CD30 molecule was lower (5 vs. 21 %). In a cohort of 31 XLA patients, plasma levels of soluble (s)LAG-3 and sCD30, chosen as indirect indicators of the Th1 / Th2 activity in vivo, were significantly higher and lower, respectively, than those measured in 31 healthy controls. Likewise, plasma levels of interferon-inducible protein 10 and of macrophage-derived chemokine in XLA patients were significantly higher and lower, respectively, than in healthy controls. PMID- 11433391 TI - Differential effects of TNF and LTalpha in the host defense against M. bovis BCG. AB - Signaling via TNF receptor type 1 (TNFR1) was shown to be crucial in host defense against the intracellular pathogens L. monocytogenes, M. tuberculosis and M. bovis. To investigate the function of TNF and LTalpha in host defense against M. bovis, mice double deficient for TNF and LTalpha (TNF / LTalpha (- / -)), TNF / LTalpha (- / -) mice complemented with a murine LTalpha transgene (TNF(- / -)) and LTalpha (- / -) mice were infected with BCG and the ensuing pathology was investigated. Control mice showed a normal host defense with early clearance of bacteria. The granulomatous reaction in the liver was accompanied by recruitment of activated macrophages characterized by their acid phosphatase positivity and differentiation into epithelioid cells as well as a coordinated expression of proinflammatory transcripts. In contrast, TNF / LTalpha (- / -) mice showed no comparable recruitment of activated macrophages in the liver. Furthermore, these mice showed extensive necrotic pulmonary lesions with massive growth of acid fast bacilli. Reintroduction of LTalpha as a transgene into TNF / LTalpha (- / -) mice prolonged survival but did not restore resistance to BCG. This, at least partially protective role of LTalpha was further supported by data demonstrating that LTalpha -deficient mice as well were susceptible to BCG infection. In contrast to the deleterious effect of TNF / LTalpha deficiency in BCG infection, BCG-infected TNF / LTalpha (- / -) mice were tolerant to LPS-induced shock. These results demonstrate that TNF as well as LTalpha are involved in murine host defense against BCG and that absence of TNF / LTalpha protects BCG-infected mice from LPS mediated shock. PMID- 11433392 TI - MHC class Ia-restricted T cells partially account for beta2-microglobulin dependent resistance to Mycobacterium tuberculosis. AB - Recent studies have highlighted the heterogeneous nature of the CD8(+) T cell response during human Mycobacterium tuberculosis infection; MHC class Ia, MHC class Ib and CD1 have all been identified as significant restriction elements. Here we have attempted to define the role of MHC class Ia in resistance to M. tuberculosis infection in mice. The course of M. tuberculosis infection in mice deficient in a single MHC class Ia molecule, either H2-K(b) or H2-D(b), was essentially identical to that observed in wild-type mice. In contrast, mice fully deficient in MHC class Ia molecules (H2-K(b) / H2-D(b) double knockout mice) were substantially more susceptible to M. tuberculosis infection. However, the double knockout mice were not as susceptible as beta 2-microglobulin-deficient mice, which have a broader phenotypic deficit. Thus, antigen presentation via MHC class Ia is an important component in resistance to M. tuberculosis, but its absence only partially accounts for the increased susceptibility of beta 2-microglobulin deficient mice. PMID- 11433393 TI - Thiolutin, an inhibitor of HUVEC adhesion to vitronectin, reduces paxillin in HUVECs and suppresses tumor cell-induced angiogenesis. AB - Recent studies have shown that integrin alpha v beta 3, a receptor for vitronectin, plays an important role in tumor-induced angiogenesis and tumor growth and that antagonists of alpha v beta 3 inhibit angiogenic processes including endothelial cell adhesion and migration. On the other hand, most inhibitors of integrin alpha v beta 3 are peptide antagonists that include the Arg-Gly-Asp (RGD) motif. We therefore reasoned that non-peptide inhibitors of endothelial cell adhesion to vitronectin might be useful for inhibition of tumor angiogenesis in vivo. We screened for low-molecular-weight natural products able to inhibit adhesion of human umbilical vein endothelial cells (HUVECs) to vitronectin, and pyrrothine group compounds including aureothricin, thioaurin and thiolutin were isolated from microbial culture broths. Of these compounds, thiolutin inhibited adhesion of HUVECs to vitronectin the most effectively (IC(50), 0.83 microM). In vivo experiments showed that thiolutin significantly suppressed angiogenesis induced by tumor cells (S-180), a pathological form of neovascularization, in a mouse dorsal air sac assay system. To explore the mechanism of inhibition of HUVEC adhesion to vitronectin by thiolutin, we examined the effect of this agent on intracellular cell adhesion signaling. We found that the amount of paxillin in HUVECs was significantly reduced by thiolutin treatment, while those of other focal adhesion proteins including vinculin and focal adhesion kinase (FAK) were not. Metabolic labeling experiments showed that thiolutin enhanced degradation of paxillin in HUVECs. Protease inhibitors (MG115 and E64-D) decreased the rate of degradation of the paxillin induced by thiolutin and partially restored thiolutin-induced inhibition of HUVEC adhesion to vitronectin. Based on these findings, we concluded that thiolutin, an inhibitor of HUVEC adhesion to vitronectin, reduces the paxillin level in HUVECs and suppresses tumor cell-induced angiogenesis in vivo. PMID- 11433394 TI - Suppressed gap junctional intercellular communication in carcinogenesis of endometrium. AB - To examine whether and at which stage of endometrial carcinogenesis decreased connexin expression occurs, we investigated changes in the expression of the gap junction proteins, connexin 26 (Cx26), Cx32 and Cx43, in human endometrial hyperplasia and cancer samples. Forty-eight endometrial tissue samples (15 endometrial hyperplasias and 33 endometrial cancers) were subjected to immunofluorescence and RT-PCR analysis. In endometrial hyperplasia, Cx26 was aberrantly expressed in all samples as revealed immunohistochemically. There was weak or negative expression in 12 samples (80.0%) and diffuse expression in cytoplasm in 3 samples (20.0%). Cx32 expression in those samples was similar to that of Cx26; there was weak or negative expression in 11 samples (73.3%) and diffuse expression in 4 samples (26.7%). In endometrial cancer, Cx26 was expressed weakly or negatively in 25 samples (75.8%), diffusely in 6 samples (18.2%) and normally in 2 samples (6.1%), while Cx32 was expressed weakly or negatively in 26 samples (78.8%), diffusely in 5 samples (15.2%) and normally in 2 samples (6.1%). It was confirmed that weak staining of Cx26 and Cx32 was due to poor expression of their mRNA. All samples showed weak Cx43 protein expression as revealed by immunohistochemical analysis. In the majority of samples, concomitant expression levels of Cx26 and Cx32 protein were observed, confirming our long term hypothesis that Cx26 and Cx32 are both abnormally regulated in a coordinated fashion in the endometrium. Our results indicate that during endometrial carcinogenesis, loss of gap junctional intercellular communication (GJIC) may occur due to the suppressed expression and the aberrant localization of connexin at relatively early stages. PMID- 11433395 TI - Hybrids of pluripotent and nullipotent human embryonal carcinoma cells: partial retention of a pluripotent phenotype. AB - To investigate whether the failure of human EC cells that do not differentiate is due to the loss of key differentiation-permissive functions or the acquisition of specific inhibitory functions, we tested the ability to differentiate of 2 hybrids produced between a relatively nullipotent human EC cell line, 2102Ep, and a pluripotent human EC cell line NTERA2. Both hybrids, which exhibited an EC phenotype, were able to differentiate readily in response to retinoic acid. Furthermore, 1 hybrid produced a well-differentiated xenograft tumor, which contained, like the NTERA2 tumors, glandular structures, loose mesenchymal tissues and nodules of cartilage, after injection into a SCID mouse. Thus, the failure of 2102Ep EC cells to differentiate is recessive and due to the loss of a key gene function or functions. Nevertheless, the hybrids differed from the pluripotent NTERA2 line by failing to differentiate in neurons, indicating that 2102Ep cells also had acquired a specific, dominantly-acting, inhibitory mutation specific to the neural lineage. Furthermore, the expression of collagen II by one hybrid before and after induction with retinoic suggested a propensity for spontaneous differentiation not evident in the parental NTERA2 cells. Thus, the mechanisms that restrict the differentiation capacity of the nullipotent 2102Ep line are complex and include both recessive and dominant acting factors. PMID- 11433396 TI - Rb-associated protein 46 (RbAp46) suppresses the tumorigenicity of adenovirus transformed human embryonic kidney 293 cells. AB - The retinoblastoma suppressor (Rb)-associated protein 46 (RbAp46) is a nuclear protein of the WD-repeat protein family and a component of the histone deacetylase complex that physically interacts with Rb. We demonstrated that RbAp46 is a gene up-regulated by the Wilms' tumor suppressor (WT1) and functions as a negative regulator of cell growth. Here we have investigated the ability of RbAp46 to inhibit malignant phenotype of adenovirus-transformed human embryonic kidney (HEK) 293 cells in tumorigenesis assays. We have found that expression of RbAp46 suppressed clonal growth of HEK 293 cells in soft agar and inhibited tumor growth of these cells in nude mice. Furthermore, expression of RbAp46 resulted in an increase of cells in the G2/M fraction of cell cycle and augmented apoptosis in serum-starved cells. The results suggest that high levels of RbAp46 expression inhibit the transformation of tumor cells through interfering with normal cell cycle and/or enhancing apoptotic cell death. PMID- 11433397 TI - Reduced expression of hepatocyte growth factor activator inhibitor type 2/placental bikunin (HAI-2/PB) in human glioblastomas: implication for anti invasive role of HAI-2/PB in glioblastoma cells. AB - Hepatocyte growth factor activator inhibitor type-2/placental bikunin (HAI-2/PB) is a serine proteinase inhibitor that contains 2 Kunitz-domains and a presumed transmembrane domain. It has broad inhibitory spectra against various serine proteinases showing potent inhibitory activities not only to hepatocyte growth factor activator but also to plasmin, trypsin and kallikreins. In this study, we investigated the expression of HAI-2/PB in human gliomas in vivo and the effects of HAI-2/PB on the fibrinolytic and invasive capabilities of human glioblastoma cells in vitro. With RNA blot analysis, HAI-2/PB mRNA was expressed in normal brain and in low-grade astrocytomas, but was hardly detectable in anaplastic astrocytomas and glioblastomas, indicating that its expression levels were inversely correlated with the histological grade of human gliomas. To further explore the possible role of HAI-2/PB in glioma progression, cultured human glioblastoma cell lines (U251 and YKG-1) were transiently transfected with an expression vector harboring human HAI-2/PB cDNA. Subsequent analysis indicated that the expression of HAI-2/PB suppressed the fibrinolytic activities of both glioblastoma cell lines. Moreover, HAI-2/PB inhibited Matrigel invasion of U251 and YKG-1 cells by 30% and 64%, respectively. This anti-invasive effect appeared to be mediated primarily by the inhibitory activity of HAI-2/PB against the serine proteinase-dependent matrix degradation. These findings suggest that the reduced expression of HAI-2/PB is possibly involved in the progression of human gliomas. PMID- 11433398 TI - Intratumor genetic heterogeneity in advanced human colorectal adenocarcinoma. AB - Colorectal carcinogenesis is widely accepted as one of the best-characterized examples of stepwise progression. The existing colorectal carcinogenesis model assumes genetic homogeneity of individual tumors for the main known genetic alterations: K-ras and p53 genes point mutations and loss of heterozygosity (LOH) of chromosome 5q and 18q. The object of the present study was to demonstrate the existence of an intratumor genetic heterogeneity in advanced sporadic colorectal carcinoma for these genetic alterations. Using improved tissue microdissection and DNA extraction, for each tumor, amplifiable DNA was obtained from 15 to 20 areas, of which 1 to 2 concerned lymph node metastases (LNM). This study revealed that 10 of 15 (67%) analyzed tumors were heterogeneous for at least 1 genetic alteration, with between 2 and 6 genotypically different clones detected per tumor. No correlation was observed between the genotype of these subclones and histological differentiation or invasive propensity. Intratumor heterogeneity was more frequently observed for LOH than for point mutations, 67% and 58% for LOH at APC and DCC locus, and 20% for mutation of either the K-ras or p53 gene. In 5 of the 9 (56%) heterogeneous cases with available LNM, the genotype observed in the LNM was different from that of the main clone in the primary tumor, and moreover, 2 of the LNM displayed a genotype undetected in the primary tumor. In conclusion, intratumor genetic heterogeneity was demonstrated in advanced sporadic colorectal carcinoma and was represented as topographically distinct genotypic subclones. Taking into account such a significant genetic heterogeneity of colorectal tumors, the use of genetic markers for prognosis management should be reconsidered. PMID- 11433399 TI - High frequency of microsatellite instability in young patients with head-and-neck squamous-cell carcinoma: lack of involvement of the mismatch repair genes hMLH1 AND hMSH2. AB - The most prevalent risk factors in the development of head-and-neck squamous-cell carcinoma (HNSCC) are excessive tobacco and alcohol consumption. In young patients with HNSCC, these risk factors are often absent. Our purpose was to investigate the risk factors, microsatellite instability (MSI) changes and status of the mismatch repair genes hMLH1 and hMSH2 in a cohort of young patients with HNSCC. Fifty-seven HNSCC tumors were examined for the presence of MSI at 16 microsatellite sites using PCR. In the young patient group (24 cases, < or = 44 years old), 100% of tumors had MSI at 1 site at least and 88% had MSI at 2 or more loci. In older patients (33 cases, > or = 45 years), MSI at 1 or more sites was found in 61% of tumors (young vs. old, p = 0.0003) and instability at 2 or more sites was found in 36% of tumors (young vs. old, p = 0.0001). The involvement of the mismatch repair genes was investigated by examining promoter methylation, exon mutation and gene expression of hMLH1 and hMSH2. All results were negative, indicating that inactivation of these 2 genes does not play a role in the development of MSI in tumors from this patient group. Furthermore, the young patient group had a significantly lower incidence of smoking (46% young, 88% old; p = 0.001) and alcohol consumption (33% young, 67% old; p = 0.0169), emphasizing the probable importance of other environmental and/or genetic factors in the development of their disease. PMID- 11433400 TI - CDKN2A is not the principal target of deletions on the short arm of chromosome 9 in neuroendocrine (Merkel cell) carcinoma of the skin. AB - The majority of small-cell lung cancers (SCLCs) express p16 but not pRb. Given our previous study showing loss of pRb in Merkel cell carcinoma (MCC)/neuroendocrine carcinoma of the skin and the clinicopathological similarities between SCLC and MCC, we wished to determine if this was also the case in MCC. Twenty-nine MCC specimens from 23 patients were examined for deletions at 10 loci on 9p and 1 on 9q. No loss of heterozygosity (LOH) was seen in 9 patients including 2 for which tumour and cell line DNAs were examined. Four patients had LOH for all informative loci on 9p. Ten tumours showed more limited regions of loss on 9p, and from these 2 common regions of deletion were determined. Half of all informative cases had LOH at D9S168, the most telomeric marker examined, and 3 specimens showed loss of only D9S168. A second region (IFNA-D9S126) showed LOH in 10 (44%) cases, and case MCC26 showed LOH for only D9S126, implicating genes centromeric of the CDKN2A locus. No mutations in the coding regions of p16 were seen in 7 cell lines tested, and reactivity to anti p16 antibody was seen in all 11 tumour specimens examined and in 6 of 7 cell lines from 6 patients. Furthermore, all cell lines examined reacted with anti p14(ARF) antibody. These results suggest that neither transcript of the CDKN2A locus is the target of deletions on 9p in MCC and imply the existence of tumour suppressor genes mapping both centromeric and telomeric of this locus. PMID- 11433401 TI - Survival of breast cancer patients in BRCA1, BRCA2, and non-BRCA1/2 breast cancer families: a relative survival analysis from Finland. AB - Reports on the prognosis of familial breast cancer patients have been contradictory. True differences in survival, if they exist, would have important implications for genetic counselling and in treatment of hereditary breast cancer. We assessed the survival rates of 359 familial breast cancer patients (32 patients from BRCA1-positive families, 43 patients from BRCA2-positive families and 284 patients from BRCA1/2-negative breast cancer families) and compared them with those of all other breast cancer patients diagnosed in Finland from 1953 to 1995 (n = 59,517). Cumulative relative survival rates (RSR) were calculated by dividing the observed survival rates by the expected ones. The expected survival rates were derived from the sex, age and calendar year specific life-tables of the general population in Finland. Regression model was used to calculate relative excess risk of death (RR) and to adjust for confounding factors. The overall 5-year RSR of the patients in the BRCA1 families, BRCA2 families, non BRCA1/2 families and among sporadic cases was 67%, 77%, 86% and 78%, respectively. However, we found no significant differences in the RR adjusted for age, stage and year of diagnosis between the different familial patient groups or the general breast cancer population. In the BRCA1 families the RR tended to be higher [RR 1.30, 95% confidence interval (CI) 0.63--2.70] and in the BRCA2 families lower (RR 0.78, 95% CI 0.39--1.57) than among the general breast cancer patient population. The RR among patients in the non-BRCA1/2 families did not differ from that of the general patient population. PMID- 11433402 TI - Inactivation of O(6)-methylguanine-DNA methyltransferase by glucose-conjugated inhibitors. AB - The DNA-repair protein O(6)-methylguanine-DNA methyltransferase (MGMT) is a decisive determinant of resistance of tumor cells to methylating and chloroethylating anti-cancer drugs. Therefore, selective inhibition of MGMT in tumors is expected to cause tumor sensitization. Several inhibitors of MGMT have been developed which function in both tumors and normal tissue. To deplete MGMT preferentially in tumors, strategies to target the inhibitor to the tumor tissue need to be developed. Here, we report on the properties of glucose-conjugated MGMT inhibitors that might be useful for tumor targeting since tumor cells frequently over-express glucose transporter. O(6)-Benzylguanine (O6BG), 8-aza O(6)-benzylguanine, O(6)-(4-bromothenyl)-guanine (O6BTG) and the corresponding spacer-linked beta-D-glucose conjugates were analyzed comparatively for MGMT inhibitory activity. Substitution at the N9 position of the purine moiety resulted generally in a reduction in the efficiency with which the inhibitors blocked MGMT. However, the inhibitory activity of the O6BTG conjugates increased with increasing spacer length, and O6BTG conjugated with a C8 spacer with beta-D glucose was nearly as effective as O6BTG on its own. MGMT was inhibited by the conjugates both in crude cell extracts and upon treatment of intact HeLa cells, indicating efficient uptake of the glucose conjugates into cells. Since the O6BTG C8-D-glucose conjugate 8-[O(6)-(4-bromothenyl)-guan-9-yl]-octyl-beta-D-glucoside was highly efficient at MGMT inhibition in a non-toxic concentration range, the drug might be a useful tool for specific tumor sensitization. PMID- 11433403 TI - Dietary intake and resting energy expenditure in relation to weight loss in unselected cancer patients. AB - Weight loss and anorexia are frequent findings in advanced cancer. The progressive wasting could be attributed to changes in dietary intake and/or energy expenditure mediated by metabolic alterations. In this study, we analyzed dietary intake in generalized malignant disease of solid tumor type in relation to resting energy expenditure (REE) and reported weight loss. In a group of 297 unselected cancer patients from a university hospital outpatient clinic, dietary intake of energy and macronutrients from a 4-day food record, REE by indirect calorimetry, height, weight and weight loss were recorded. Protein intake was validated against 24 hr urine nitrogen in a subgroup (n = 53), and no indication of systematic misreporting was found. Mean daily dietary intake was below maintenance requirements, 26 +/- 10 kcal/kg. Weight loss of more than 10% was present in 43% of patients and elevated REE (>110% of predicted) in 48%. Dietary intake did not differ between normo- and hypermetabolic patients, nor was tumour type or gender related to energy and protein intake. Weight loss could not be accounted for by diminished dietary intake since energy intake in absolute amounts was not different and intake per kilogram body weight was higher in weight-losing patients compared to weight-stable patients. Dietary macronutrient composition did not differ from the general population. Dietary intake of energy and protein was decreased, but dietary macronutrient composition did not appear to be changed. Weight loss and hypermetabolism were frequent and not compensated for by an increase in spontaneous food intake. Our results indicate that an expected up-regulation of dietary intake in response to elevated energy expenditure is frequently lost in cancer patients. This may be the explanation behind cancer cachexia rather than a primary decrease in appetite. PMID- 11433404 TI - Chloroaluminum phthalocyanine tetrasulfonate delivered via acid-labile diplasmenylcholine-folate liposomes: intracellular localization and synergistic phototoxicity. AB - Folate-diplasmenylcholine (1,2-di-O-(Z-1'-hexadecenyl)-sn-glycero-3 phosphocholine; DPPlsC) liposomes have been shown to greatly enhance the potency of water-soluble antitumor agents via a selective folate-mediated uptake and acid catalyzed endosomal escape mechanism (Rui et al. J. Am. Chem. Soc., 1998; 120:11213--18). This study describes an adaptation of this strategy for the delivery of chloroaluminum phthalocyanine tetrasulfonate ([AlPcS(4)](4-)), a water-soluble sensitizer used in photodynamic therapy, in a binary targeting scheme designed to enhance both its tumor selectivity and phototoxicity. [AlPcS(4)](4-)/DPPlsC:folate liposomes (9.8 microM bulk concentration, 2.5 mM intraliposomal concentration) were substantially more phototoxic to folate deficient KB cells than 12.5 microM free [AlPcS(4)](4-) after a 30 min irradiation (630-910 nm). Considerable differences in phototoxicity were observed, however, between the commercially-available AlPcS(4)(4-) and an HPLC purified sample of [AlPcS(4)](4-) due to an increased tendency for the latter to aggregate. Experiments with [AlPcS(4)](4-)/DPPC:folate and folate-free [AlPcS(4)](4-)/DPPlsC liposomes (acid-insensitive and non-targeted controls, respectively) showed significantly reduced phototoxicities under the same illumination conditions. Our results imply that higher concentrations of water soluble sensitizers can be delivered to target cells using the folate receptor mediated pathway, which can change both the biodistribution and intracellular localization of the sensitizer when acid-labile DPPlsC liposomes are used as the delivery vehicle. Potential advantages of this approach include the use of lower bulk [AlPcS(4)](4-) concentrations, rapid plasma clearance of free [AlPcS(4)](4 ), and better phototoxic responses, due to higher intracellular [AlPcS(4)](4-) concentrations combined with reduced collateral photodamage arising from misguided sensitizer accumulation, thereby enhancing the selective phototoxicity of PDT treatments. PMID- 11433405 TI - Gene delivery using a receptor-mediated gene transfer system targeted to hepatocellular carcinoma cells. AB - For gene therapy to be effective in cancers, it is necessary to deliver therapeutic genes into cells with high specificity and efficiency. In this study, we examined the in vitro and in vivo gene delivery efficiency of a new, growth receptor-mediated gene transfer system in hepatocellular carcinoma (HCC). The effects of transfection of wild-type p53 using this system were also studied. The system consisted of a ligand oligopeptide for epidermal growth factor receptor (EGFR) recognition, a polypeptide for DNA binding, and an endosome-releasing oligopeptide for endosomolysis. Two human HCC cell lines and a normal liver cell line were used, and pCMV-beta-galactosidase (beta-gal) was used as a reporter gene. Both HCC cell lines had strong expression of EGFR and the in vitro transfer efficiency peaked at day 5 at about 50%. This finding was in contrast to the normal liver cell line, which had weak EGFR expression and less than 1% transfer efficiency throughout. For in vivo gene transfer in tumors produced by inoculating HCC cells in nude mice and with the vector-beta-gal gene complex injected peritumorally, beta-gal expression was detected within the tumors at 12 hr, peaked at day 5 involving about 50% of the tumor cells and persisted at 2 weeks. Using this vector system, transfection of wild-type p53 into Huh-7 cells that had mutated p53 resulted in significant growth inhibition of cancer cells accompanied by a decreased G2/M phase and increased p53 protein. In conclusion, this receptor-mediated gene transfer system appears to work specifically in HCC cells with high efficiency, and may be promising in delivering apoptotic and other genes into HCC cells. PMID- 11433406 TI - Cell cycle effects of gemcitabine. AB - Gemcitabine (2',2'-difluoro-2'-deoxycytidine, or dFdC) is a promising anticancer agent with demonstrated clinical activity in solid tumours currently undergoing clinical trials. Despite extensive studies on the biochemical mechanism of action, cell cycle perturbations induced by dFdC have not yet been thoroughly investigated, apart from the expected inhibition of DNA synthesis. The aim of our study was to clarify whether cell population kinetics is a vital factor in the cytotoxicity of dFdC in single or repeated treatments and in the dFdC-cisplatin combination. Ovarian cancer cells growing in vitro were treated with dFdC for 1 hr in a range of concentrations from 10 nM to 10 microM. Cell kinetics was investigated by DNA-bromodeoxyuridine flow cytometry, using different experimental protocols to measure either the time course of DNA-synthesis inhibition or the fate of cells in G(1), S or G(2)M at the time of dFdC treatment or 24 hr later. A modified sulforhodamine B test was used to assess the growth inhibition caused by dFdC given alone or with cisplatin. Although dFdC promptly inhibited DNA synthesis, cytotoxicity on proliferating cells was not specific for cells initially in the S phase. DNA synthesis was restored after a G(1) block of variable, dose-dependent length, but recycling cells were intercepted at the subsequent checkpoints, resulting in delays in the G(2)M and G(1) phases. The activity of repeated treatment with dFdC + dFdC or dFdC + cisplatin was highly dependent on the interval length between them. These results suggest that the kinetics of cell recycling from a first dFdC treatment strongly affects the outcome of a second treatment with either dFdC itself or cisplatin. PMID- 11433407 TI - Bispecific single-chain antibodies as effective tools for eliminating epithelial cancer cells from human stem cell preparations by redirected cell cytotoxicity. AB - High-dose chemotherapy (HDC) with autologous bone marrow or peripheral stem cell transplantation is discussed as one option to treat the extensive stage of a variety of tumors. Effective methods to eliminate contaminating tumor cells from human bone marrow or stem cell grafts may improve the outcome of the patients. We investigated 3 recombinant bispecific single-chain antibodies (bscAbs) directed against 17-1A (EpCAM), c-erbB-2 (HER-2/neu) and LeY on the one and CD3 on the other binding site for their ability to induce lysis of epithelial tumor cells by retargeting autochthonous T lymphocytes present in bone marrow mononuclear cells (BMMC) and in peripheral stem cell mononuclear cells (PSMC). The bscAbs showed remarkable specific lysis of different epithelial tumor cell lines with BMMCs as well as with PSMCs as effector cells. Investigation of the alpha 17-1A-alpha CD3 bscAb revealed a significant correlation between the percentage of CD3(+) cells present in the BMMCs and the rate of lysis as well as the absence of detrimental effects on the viability of hematopoietic progenitor cells as determined by colony-forming unit assays (CFUs). Our results indicate that recombinant bispecific single-chain antibodies could be new tools for purging of human bone marrow and peripheral stem cell grafts from contaminating epithelial cancer cells for patients receiving autologous stem cell transplantation after HDC. PMID- 11433408 TI - A case-control study of gastric cancer in Venezuela. AB - A case-control study to evaluate risk factors for gastric cancer was carried out among 292 cases of gastric cancer and 485 controls in a high-risk area of Venezuela. Subjects were interviewed using a structured questionnaire, which elicited information on residential history, socio-economic status, family history of gastric diseases, smoking, drinking and dietary habits. Habitual diet was estimated from a meal-structured food frequency questionnaire on 75 food items. There was a strong inverse association with social class, as measured by education and by indicators of poverty. The results of the dietary analysis suggest that a diet high in starch and low in meat, fish and fresh vegetables increases risk of gastric cancer. A protective effect was observed for frequent consumption of allium vegetables. Inverse associations were found with height, which may reflect nutritional status in childhood, and with refrigerator use in the first two decades of life. Alcohol and tobacco consumption was investigated among males only, since the prevalence of alcohol and tobacco use was very low in females. Alcohol drinkers were at higher risk than non-drinkers and there was a small excess risk for current smokers compared with never smokers. There was some evidence of familial aggregation of gastric cancer. These findings will have important implications in planning preventive strategies for gastric cancer in Venezuela. PMID- 11433409 TI - Molecular detection of cervical intraepithelial neoplasia and cervical carcinoma by microsatellite analysis of Papanicolaou smears. AB - Carcinoma of the uterine cervix is one of the most common malignancies worldwide, yet it is clearly preventable by population screening. The Papanicolaou (Pap) smear has proved to be the most successful test for the detection of precancerous lesions and is largely responsible for the reduction of cervical cancer mortality and morbidity rates. However, the Pap smear is not perfect; false-negative results of various rates are reported. To improve the diagnostic efficacy of cervical cytology, we performed microsatellite analysis on paired Pap smear samples from cervical lesions. Nine microsatellite markers were chosen from chromosomal regions commonly displaying loss of heterozygostity (LOH) in cervical cancer and those displaying microsatellite instability (MI) in other squamous cell cancer. Microsatellite alterations were detected in 16/21 (76%) Pap smear DNA samples including 11 of 13 (85%) smears from invasive squamous cell carcinomas (SCCs) and 5 of 8 (63%) from squamous intraepithelial lesions (SILs). Microsatellite alterations detected in the Pap smear DNA were identical to those identified in seven paired primary tumors available for analysis. Moreover, this molecular approach detected genetic alterations in two cases apparently negative by cytologic examination. None (0/25) of the control patients displayed microsatellite alterations in paired Pap smears. Microsatellite analysis of cervical cytologic samples may provide a complementary method to analyze suspicious but not diagnostic cytologic samples further. PMID- 11433410 TI - Non-Hodgkin lymphoma among young adults with and without AIDS in Italy. AB - To compare the presentation and prognosis of non-Hodgkin lymphoma (NHL) in people with AIDS (PWA) and in the general Italian population, a record linkage study was carried out. The fraction of NHLs attributable to HIV/AIDS was also estimated. Information from the National AIDS Registry (RAIDS) was linked with records from 13 cancer registries (CR), covering about 15% of the Italian population. During the period 1985--94, among PWA ages 15--49, 136 NHLs were identified (8% of all NHLs) and were compared with 1,481 concurrent incident NHL cases of the same age group among non-PWA. Percentages above 13% of all NHLs were registered in the northern areas of Genoa and Varese, i.e., the most heavily affected by the AIDS epidemic. Between 1 year prior to and 3.5 years after AIDS diagnosis, PWA showed an overall standardised incidence ratio (SIR) for NHL of 302. SIR was particularly high (394) within 3 months after AIDS diagnosis and subsequently declined to 170. SIR was somewhat higher in females (428) than in males (280) but similar among intravenous-drug users (299) and other HIV-transmission groups (309). High-grade NHL, particularly immunoblastic and Burkitt's lymphoma, were twice as frequent among PWA than non-PWA. Conversely, low-grade NHL were less frequent. Except for the high proportion of brain localisation, no clear difference emerged in the pattern of NHL presentation site in PWA compared with non-PWA. At variance with NHL in the general population, among PWA histological grade had little impact on survival, which overall appeared to be very poor (2 year survival: 10%; 95% confidence interval: 3%--17%). Our present linkage of RAIDS and CRs represents an efficient tool for the surveillance of trends in incidence and survival of NHL among PWA in Italy. PMID- 11433411 TI - In vitro BPDE-induced DNA adducts in peripheral lymphocytes as a risk factor for squamous cell carcinoma of the head and neck. AB - The level of DNA adducts under the same conditions of carcinogen exposure and cell proliferation reflects an integrated measure of carcinogen metabolism and DNA repair. Therefore, such DNA adduct levels have the potential to be a biomarker for susceptibility to chemical carcinogenesis. In a pilot study of 91 patients with squamous cell carcinomas of the head and neck and 115 controls who were frequency matched by age, sex, ethnicity, and smoking status, we applied a newly developed in vitro assay of benzo[a]pyrene diol epoxide (BPDE)-induced DNA adducts in short-term peripheral lymphocytes cultures. Levels of BPDE-DNA adducts were found to be significantly higher in cases than in controls (mean +/- SD, 76.8 +/- 77.4/10(7) and 47.1 +/- 48.0/10(7) nucleotides, respectively; p < 0.001). Using the median level of control values (35/10(7)) as the cut-off point, about 66% of cases were distributed above this level. Logistic regression analysis revealed that the level of BPDE-induced DNA adducts was an independent risk factor (odds ratio = 2.22; 95% confidence interval = 1.22--4.04) after adjustment for age, sex and smoking status. Further stratified analyses showed that levels of the induced adducts between cases and controls were significantly higher in both age groups, that is, younger or older than 60, as well as in both men and women. Smoking had a positive effect on the induced adducts. The highest level of induced adducts was seen in current smokers, then former smokers and non smokers. There was a statistically significant dose--response relationship between the quartile levels of BPDE-induced DNA adducts and the risk of head and neck cancer (trend test, p = 0.003). Despite the relatively small sample size, the association of BPDE-induced DNA adducts and cancer risk suggests that this assay has the potential to complement with other biomarkers in identifying individuals at increased risk of developing tobacco-related cancers. PMID- 11433412 TI - Inhibition by d-limonene of experimental hepatocarcinogenesis in Sprague-Dawley rats does not involve p21(ras) plasma membrane association. AB - The effects of d-limonene on hepatocarcinogenesis induced by N-nitrosomorpholine (NNM) and on membrane-associated p21(ras) and labeling and apoptotic indices of the liver were investigated in male Sprague-Dawley rats. Rats were given drinking water containing NNM for 8 weeks, and from the beginning of experimental week 9, they received chow pellets containing 1% or 2% limonene. The preneoplastic and neoplastic liver lesions (cellular alteration foci, neoplastic nodules and hepatocellular carcinomas), and hepatic foci staining positive for glutathione-S transferase, placental type (GST-P) were examined microscopically and histochemically. At week 16, quantitative histologic analysis showed that oral administration of 1% or 2% limonene resulted in significant reductions in the number and mean area of GST-P-positive hepatic foci and the number of cellular alteration foci, neoplastic nodules and hepatocellular carcinomas. Limonene, at both doses, also caused significant decreases in the labeling indices and significant increases in the apoptotic indices of cellular alteration foci, neoplastic nodules, hepatocellular carcinomas and adjacent liver. However, limonene, at both doses, had no significant influence on the production of membrane-associated p21(ras) in the visible liver white nodules. These findings indicate that limonene inhibits hepatocarcinogenesis and suggest that this effect may be clearly related to its effect in inhibiting cell proliferation and in enhancing apoptosis, but not through ras oncoprotein plasma membrane association. PMID- 11433413 TI - Somatic mutations of WNT/wingless signaling pathway components in primitive neuroectodermal tumors. AB - Primitive neuroectodermal tumors (PNETs) represent the most frequent malignant brain tumors in childhood. The majority of these neoplasms occur in the cerebellum and are classified as medulloblastomas (MB). Most PNETs develop sporadically; however, their incidence is highly elevated in patients carrying germline APC gene mutations. The APC gene encodes a central component of the WNT/wingless developmental signaling pathway. It regulates the levels of cytoplasmic beta-catenin protein that plays a central role in neural development and cell proliferation. We analyzed 87 sporadic PNETs and 10 PNET cell lines for mutations of the APC gene and beta-catenin (CTNNB1) gene using single strand conformational polymorphism (SSCP) and sequencing analysis. We examined the mutation cluster region of APC (codons 1255--1641) for germline variants and somatic mutations. The medulloblastoma cell line MHH-MED-2 carried a Glu1317Gln missense germline variant and a sporadic MB sample showed a somatic Pro1319Leu substitution. Mutational analysis of exon 3 of CTNNB1 uncovered 4 PNETs (4.8%) with somatic missense mutations. These mutations caused amino acid substitutions in 3 of 80 medulloblastomas (Ser33Phe, Ser33Cys and Ser37Cys) and 1 of 4 supratentorial PNETs (Gly34Val). All mutations affected GSK-3 beta phosphorylation sites of the degradation targeting box of beta-catenin and resulted in nuclear beta-catenin protein accumulation. Deletions of CTNNB1 were not detected by PCR amplification with primers spanning exons 1--5. Our data indicate that inappropriate activation of the WNT/wingless signaling pathway by mutations of its components may contribute to the pathogenesis of a subset of PNETs. PMID- 11433414 TI - DNA adduct formation by the ubiquitous environmental contaminant 3 nitrobenzanthrone in rats determined by (32)P-postlabeling. AB - Diesel exhaust is known to induce tumors in animals and is suspected of being carcinogenic in humans. Of the compounds found in diesel exhaust and in airborne particulate matter, 3-nitrobenzanthrone (3-NBA), is a particularly powerful mutagen. We investigated the capacity of 3-NBA to form DNA adducts in vivo that could be used as agent-specific biomarkers of exposure. Female Sprague-Dawley rats were treated orally with 2 mg/kg body weight of 3-NBA, and DNA from various organs was analyzed by (32)P-postlabeling. High levels of 3-NBA-specific adducts were detectable in all organs. Both enrichment versions nuclease P1 digestion and n-butanol extraction resulted in patterns consisting of either 3 or 4 adducts remarkably similar in all tissues examined. The highest level of DNA adducts was found in the small intestine (38 adducts per 10(8) nucleotides) followed by forestomach, glandular stomach, kidney, liver, lung and bladder. To provide information on the nature of the adducts formed in vivo in rats, DNA adducts were cochromatographed in 2 independent systems with standardized deoxyguanosine adducts and deoxyadenosine adducts produced by reaction of 3-NBA in the presence of xanthine oxidase with deoxyribonucleoside 3'-monophosphates in vitro. In both systems, each of the rat adducts comigrated either with a deoxyguanosine or a deoxyadenosine-derived 3-NBA adduct. Our results demonstrate that 3-NBA binds covalently to DNA after metabolic activation, forming multiple DNA adducts in vivo, all of which are products derived from reductive metabolites bound to the purine bases (deoxyguanosine 60% and deoxyadenosine 40%). PMID- 11433415 TI - No evidence of linkage to HPC20 on chromosome 20q13 in hereditary prostate cancer. PMID- 11433416 TI - Mixed lineage kinase (MLK) family members are not involved in androgen regulation of prostatic proliferation or apoptosis. AB - BACKGROUND: Once paracrine growth factors are secreted by androgen receptor expressing prostatic stromal cells, they diffuse across the basement membrane of glandular acini, where they bind to epithelial cell surface receptors. This binding stimulates signaling pathways that regulate both the rate of proliferation and apoptosis of prostate epithelial cells. In the present studies, the role of mixed lineage kinases (MLKs) in these signaling processes were studied using a pharmacological approach. METHODS: The indolocarbazole CEP-1347 (KT 7515) is a potent inhibitor of kinase activity of MKLs. Male rats were treated with CEP-1347 (1 mg/kg of body weight/day) to determine whether inhibition of the MLKs can prevent androgen ablation (i.e. castration) induced apoptosis of prostatic epithelial cells, using as indexes total ventral prostatic DNA content and the percentage of ventral prostatic epithelial cells whose DNA can be terminal transferase end-labeled. In addition, animals previously castrated a week earlier were treated daily with either vehicle or CEP-1347 and exogenous androgen replacement to induce the proliferative re-growth of the prostatic epithelial cells. After 1 week of treatment, the total ventral prostatic DNA content in the vehicle vs. CEP-1347 groups was compared. RESULTS: Using the National Center for Bio-Informatics data bank, MLK2, MLK3, and DLK members of the MLK family are expressed by the normal prostate. Inhibition of the MLKs with CEP-1347 did not affect the kinetics of apoptosis of prostatic epithelial cells induced by androgen ablation. In addition, such MLK inhibition did not prevent androgen replacement induced proliferative regrowth of the prostate epithelium in castrated animals. CONCLUSIONS: Signaling through the MLK family is not involved in either the androgen-induced proliferation or the androgen ablation-induced apoptosis of prostatic epithelial cell in the rat. PMID- 11433417 TI - Reduction of human prostate tumor vascularity by the alpha1-adrenoceptor antagonist terazosin. AB - BACKGROUND: We previously demonstrated that the quinazoline-derived a1 adrenoceptor antagonists doxazosin and terazosin suppress prostate cancer growth via apoptosis induction. The aim of this study was to determine the potential effect of a1-adrenoceptor antagonists on tumor vascularity of the human prostate. METHODS: A total of 34 men with benign prostatic hyperplasia (BPH) who have been on terazosin treatment (for the obstructive symptoms) were pathologically diagnosed with prostate cancer following surgery. These patients were stratified according to the length of treatment periods with terazosin into two groups, 1 week-6 months, and 6-17 months. The control group consisted of prostatectomy specimens from 25 untreated prostate cancer patients undergoing surgery for localized disease. Formalin-fixed, paraffin-embedded prostate specimens were analyzed for apoptosis (TUNEL assay), cell proliferation (Ki-67), microvessel density (MVD) (von Willebrand factor/Factor VIII), vascular endothelial growth factor (VEGF) expression, and prostate specific antigen (PSA) immunoreactivity. RESULTS: A significant induction of apoptosis was observed among cancerous prostatic epithelial cells in the terazosin-treated, as compared to the untreated prostate cancer specimens, while there was no significant change in the proliferative index of the same tumor cell populations after treatment. Furthermore, terazosin resulted in a significant decrease in prostate tissue MVD compared with the untreated group (P < 0.01), that correlated with the increased apoptotic index of the cancerous areas. Tissue PSA expression in the prostatic tumor foci was also markedly reduced after terazosin treatment, while no significant changes in VEGF expression were detected. CONCLUSIONS: These findings provide the first evidence that terazosin, a quinazoline-based a1-blocker decreases prostate tumor vascularity. Our study has significant clinical implications in identifying selected alpha1-adrenoceptor antagonists as potential anti-tumor agents with apoptotic and anti-angiogenic effects in the human prostate that can be exploited for the treatment of advanced prostate cancer. PMID- 11433418 TI - PEAZ-1: a new human prostate neoplastic epithelial cell line. AB - BACKGROUND: The generation of prostatic cell lines provides in vitro models for experimental studies of the pathogenesis of prostate carcinoma. Therefore, we established and characterized a new human prostate epithelial cell line, PEAZ-1 (prostate epithelial Arizona-1). METHODS: The PEAZ-1 cells were grown from a primary human prostate carcinoma specimen obtained from radical prostatectomy. The isolated cells were characterized by immunobiochemistry, immunohistochemistry, and tumorigenicity studies. RESULTS: PEAZ-1 cells are near diploid, tumorigenic, and androgen independent for cell growth. PEAZ-1 cells express N-cadherin, alpha- and beta-catenins, and p120 at cell-cell contacts, cytoplasmic laminin 5, vinculin, paxillin, and phosphotyrosine at focal adhesions, vimentin, and cytokeratins 8 and 18. They do not express plakoglobin, E-cadherin, and PSA, and do not form desmosomes and hemidesomomes. PEAZ-1 respond to ocadaic acid, a pro-apoptotic agent, by expression of p53. CONCLUSIONS: PEAZ-1 cells is a human prostate cancer cell line that has a number of mesenchymal characteristics. PMID- 11433419 TI - Down-regulation of prostasin serine protease: a potential invasion suppressor in prostate cancer. AB - BACKGROUND: Prostasin is a serine protease predominantly expressed in normal prostate epithelial cells. The biological function of prostasin has not been determined. METHODS: Western blot and RT-PCR analyses were used to examine the expression of prostasin in prostate cancer cell lines. Immunohistochemistry was used to evaluate prostasin protein expression in human prostate cancer. An in vitro Matrigel invasion assay was used to test the invasiveness of prostate cancer cell lines forced to express recombinant prostasin. RESULTS: Both prostasin protein and mRNA were found to be expressed in normal human prostate epithelial cells and a non-invasive human prostate cancer cell line, the LNCaP, but neither was found in invasive human prostate cancer cell lines DU-145 and PC 3. Prostasin mRNA expression was absent in invasive prostate cancer cell lines of a transgenic mouse model. Immunohistochemistry analysis showed that prostasin protein expression is down-regulated in high-grade prostate cancer. Transfection of DU-145 and PC-3 cells with a full-length human prostasin cDNA restored prostasin expression and reduced the in vitro invasiveness by 68 and 42%, respectively. CONCLUSIONS: Our data indicate that prostasin may be implicated in normal prostate biology and is able to suppress prostate cancer invasion in vitro. PMID- 11433420 TI - Effects and characterization of paracrine factors produced by human prostate stromal cells in bioassays using rat Sertoli cells, LNCaP tumor cells, and cultured prostate epithelial cells. AB - BACKGROUND: Prostatic stroma affects both proliferation and differentiation of epithelial cells but the factors involved remain poorly understood. In order to identify and characterize potential paracrine mediators, we studied the effects of human prostate fibroblast-conditioned media (PFCM) in three bioassay systems. METHODS: The first bioassay uses transferrin secretion by cultured rat Sertoli cells as an endpoint for differentiating activity. Factors active in this (heterologous) assay were compared to PModS, a mediator of mesenchymal-epithelial interactions in the testis, also produced by rat prostate stromal cells. The two other (homologous) bioassays use LNCaP tumor cells or subcultured human prostate epithelial cells (PEC) as targets. Differentiation is evaluated by prostate specific antigen (PSA) secretion and reverse transcriptase-polymerase chain reaction (RT-PCR) for a number of markers of epithelial function. Proliferation is assayed by measurements of DNA and thymidine incorporation. RESULTS: PFCM markedly stimulates transferrin production by Sertoli cells. The main factor(s) involved are acid stable and bind to heparin. However, both their size (approximately 37 kDa) and their behavior on reversed-phase chromatography differ from that of PModS. Although PFCM increases total RNA content of LNCaP, it does not increase or restore differentiated function of LNCaP or PEC. Proliferative effects are observed in LNCaP and these effects cannot be neutralized by an antiserum directed against basic fibroblast growth factor (bFGF). Antiproliferative effects are observed in PEC and these effects are largely due to transforming growth factor-beta (TGF-beta). CONCLUSIONS: PFCM provokes differentiating effects in a Sertoli cell bioassay, but unlike with rat stromal cells, the factor(s) involved differ from PModS. In the two homologous systems studied, differentiating effects could not be demonstrated and discordant effects were noted on proliferation. Various bioassay systems will be required to identify the spectrum of mediators present in PFCM. PMID- 11433421 TI - Calcium, dairy products, and the risk of prostate cancer. AB - BACKGROUND: Calcium intake has been suggested to play a role in the etiology of prostate cancer, since it is inversely related to vitamin D levels, which, in turn, would have an antiproliferative effect on human cancer cell lines. The hypothesis that high calcium and low vitamin D levels are associated with prostate cancer risk, however, remains unconvincingly demonstrated. METHODS: This relation was investigated in a case-control study of prostate cancer conducted in Northern Italy between 1985 and 1992 on 288 cases and 762 controls admitted to hospital for acute non-neoplastic diseases. RESULTS: No significant relation was found for calcium intake: compared to the lowest quintile of intake, the multivariate odds ratios (OR) of prostatic cancer, after adjusting for age, study center, education, body mass index, and meat intake, for increasing intake quintiles were 1.21, 0.68, 0.64, 1.12, with no trend in risk. The OR was 0.99 for an increment of 500 mg per day of calcium. The risk estimates were consistent in two strata of age (< 65 and > or = 65 years at diagnosis). CONCLUSIONS: Our results do not support an association between calcium and the risk of prostate cancer. However, the present study cannot address the effect of calcium in the ranges where an increased risk of prostate cancer has been noted previously, and with specific reference to advanced disease. PMID- 11433422 TI - Activation of latent protease function of pro-hK2, but not pro-PSA, involves autoprocessing. AB - BACKGROUND: Human glandular kallikrein 2 (hK2) and prostate-specific antigen (PSA) are members of an extensive kallikrein family of proteases. Both proteases are secreted as zymogens or proenzymes containing a seven amino acid propeptide that must be proteolytically removed for enzymatic activation. The physiological proteases that activate pro-hK2 and pro-PSA are not known. METHODS: The pro-hK2 peptide sequence is Val-Pro-Leu-Ile-Gln-Ser-Arg (VPLIQSR). For PSA, the amino acid sequence of the propeptide is Ala-Pro-Leu-Ile-Leu-Ser-Arg (APLILSR). Fluorescent substrates were made by coupling these peptide sequences to 7-amino-4 methylcoumarin (AMC). The hydrolysis of the VPLIQSR-AMC and APLILSR-AMC substrates by hK2, PSA, and a panel of purified proteases was determined. RESULTS: HK2 readily cleaved the pro-hK2 peptide substrate VPLIQSR-AMC with a rate of hydrolysis that was approximately 8-fold higher than an equimolar amount of purified trypsin. HK2 also had the highest hydrolysis rate from among a group of other trypsin-like proteases. In contrast, neither hK2 nor PSA was able to appreciably cleave the pro-PSA substrate APLILSR-AMC. The pro-PSA substrate was most readily hydrolyzed by urokinase and trypsin. CONCLUSIONS: HK2 can hydrolyze the pro-hK2 substrate suggesting that maturation of pro-hK2 to enzymatically active hK2 involves autoprocessing. As expected, PSA, a chymotrypsin-like protease, was unable to hydrolyze either of the propeptide substrates. Therefore, it is unlikely that PSA can auto-process its own enzymatic function. HK2 has trypsin-like specificity but was unable to hydrolyze the pro-PSA substrate. These results raise the possibility that an additional processing protease may be required to fully process PSA to an enzymatically active form. PMID- 11433423 TI - Glutamine as a pathogenic factor in hepatic encephalopathy. AB - Hepatic encephalopathy (HE) results from acute or chronic liver dysfunction and is associated with hyperammonemia. Ammonium ions penetrate from blood to brain, where they form glutamine (Gln) in the reaction with glutamate catalyzed by an astroglia-specific enzyme, glutamine synthetase (GS). Experimental data suggest that many manifestations of HE can be ascribed to increased Gln synthesis and accumulation in the brain. In HE resulting from acute liver failure ("fulminant hepatic failure"), the osmotic action of Gln appears to be in a large degree responsible for cerebral edema and edema-associated disturbances of cerebral blood flow and ionic homeostasis. In chronic HE not accompanied by cerebral edema, Gln contributes to impairment of cerebral energy metabolism, and its increased transport from brain to the periphery accelerates the blood-to-brain transport of aromatic amino acids, of which tryptophen (Trp) is converted to metabolites directly implicated in HE. Most of the evidence that Gln participates in pathological events has been derived from their disappearance or amelioration in HE rats in which the cerebral Gln content was reduced by treatment with a GS inhibitor, methionine sulfoximine. PMID- 11433424 TI - Subcellular distribution, calmodulin interaction, and mitochondrial association of the hyaluronan-binding protein RHAMM in rat brain. AB - The CNS contains high levels of the glycosaminoglycan hyaluronan, and neural cells express a variety of proteins that are members of the hyaladherin family of hyaluronan-binding proteins. We have previously shown that the hyaladherin RHAMM (receptor for hyaluronan-mediated motility; CD168) is expressed by neural cells in culture; plays a role in astrocyte motility, neurite migration, and axonal growth; and is widely distributed in neurons and oligodendrocytes of developing and adult rat CNS. Here we demonstrate differential localization of various forms of RHAMM in subcellular fractions of adult rat brain. Western blotting indicated the presence of 66, 75, and 85-90 kDa molecular weight RHAMM forms in whole-brain homogenates. Subfractionation revealed enrichment of the 66 and 85-90 kDa forms in soluble fractions, whereas the 75 kDa form was enriched in mitochondrial fractions. This latter form was retained in osmotically shocked mitochondria, but was liberated by alkali carbonate, suggesting a nonintrinsic mitochondrial membrane association. By double immunohistochemical labeling for RHAMM and the mitochondrial marker cytochrome oxidase, RHAMM was localized to isolated mitochondria in vitro and to neuronal mitochondria in vivo. Hyaluronan-sepharose chromatography and cetylpiridinium chloride precipitation confirmed the hyaluronan-binding capacity of RHAMM forms. By calmodulin-affinity chromatography, endogenously expressed brain RHAMM was demonstrated to bind calmodulin in a Ca2+-dependent manner. These results, together with reports of RHAMM association with actin and microtubules in other systems, suggest a role of RHAMM in calmodulin-mediated cell signaling to cytoskeletal elements and/or mitochondria in the CNS and invoke novel functions of its interactions with hyaluronan. PMID- 11433425 TI - Human medulloblastoma cell line DEV is a potent tool to screen for factors influencing differentiation of neural stem cells. AB - The aim of our study was to investigate whether a human neural cell line could be used as a reliable screening tool to examine the functional conservation, in humans, of transcription factors involved in neuronal or glial specification in other species. Gain-of-function experiments were performed on DEV cells, a cell line derived from a human medulloblastoma. Genes encoding nine different transcription factors were tested for their influence on the process of specification of human DEV cells towards a neuronal or glial fate. In a first series of experiments, DEV cells were transfected with murine genes encoding transcription factors known to be involved in the neuronal differentiation cascade. Neurogenins-1, -2, and -3; Mash-1; and NeuroD increased the differentiation of DEV cells towards a neuronal phenotype by a factor of 2-3.5. In a second series of experiments, we tested transcription factors involved in invertebrate glial specification. In the embryonic Drosophila CNS, the development of most glial cells depends on the master regulatory gene glial cell missing (gcm). Expression of gcm in DEV cells induced a twofold increase of astrocytic and a sixfold increase of oligodendroglial cell types. Interestingly, expression of tramtrack69, which is required in all Drosophila glial cells, resulted in a fourfold increase of only the oligodendrocyte phenotype. Expression of the related tramtrack88 protein, which is not expressed in the fly glia, or the C. elegans lin26 protein showed no effect. These results show that the Drosophila transcription factor genes tested can conserve their function upon transfection into the human DEV cells, qualifying this cell line as a screening tool to analyze the mechanisms of neuronal and glial specification. PMID- 11433426 TI - Expression of a voltage-dependent potassium channel protein (Kv3.1) in the embryonic development of the auditory system. AB - The present study traces the development of a voltage-dependent potassium channel protein (Kv3.1) in the avian homologue of the cochlear nucleus, in the cochleovestibular ganglion, and in the otic epithelium from early developmental stages until near hatching. Immunohistochemistry with antibodies to the carboxy terminus (recognizing the Kv3.1b splice variant) and to the amino terminus (recognizing either form of Kv3.1) was used on Hamburger-Hamilton-staged chicken embryos. There were three periods in the relative levels of immunostaining in these regions. Early (E2-6), when precursor cells proliferate, migrate, and form axons, there was staining when using either antibody. In the middle period (E6 11), marked by hair cell differentiation, dendritic growth, and early synapse formation, staining levels decreased. In the late period (E11-19), when auditory function begins, staining increased rapidly, especially for Kv3.1b. Early Kv3.1 expression occurs in neuronal and hair cell precursors before they differentiate or function. Later, in the otic epithelium, a high level of Kv3.1 in cilia may precede or coincide with the onset of hair cell function. In neurons, some features of its localization correlate with axon outgrowth and synapse formation, others with the onset of neural activity and function. PMID- 11433427 TI - Macrophage colony-stimulating factor is expressed in neuron and microglia after focal brain injury. AB - In a previous study, we have demonstrated that damaged neurons within a boundary area around necrosis fall into delayed neuronal death owing to the cytotoxic effect of microglial nitric oxide (NO), and these neurons are finally eliminated by activated microglia. In this process, microglia are activated to release NO, increase in number, and accumulate toward the damaged area. In this study, we investigated the expression of macrophage colony-stimulating factor (M-CSF, also called colony stimulating factor-1; CSF-1) and other cytokines, which are reported to relate to activation, proliferation, or migration of microglia. The mRNA of M-CSF arose biphasically from 30 min to 1 hr and from 6 to 72 hr after the injury, as demonstrated by semiquantitative RT-PCR. However, another cytokine of granulocyte-macrophage CSF (GM-CSF) or interleukin-3 (IL-3), which causes proliferation of microglia in vitro, was not detected. From immunohistochemical studies, positive staining of M-CSF was observed mainly in neuron-specific enolase (NSE)-positive cells from 1 to 12 hr after the injury, and after that M CSF became positive in Griffonia simplicifolia isolectin-B4 (GSA-I-B4)-positive cells from 24 to 72 hr in the boundary area around necrosis. These results suggest that neurons around the damaged area express M-CSF in the early phase after injury, which may initially activate microglia, and these activated microglia also express M-CSF later, causing further proliferation or migration of microglia themselves to eliminate damaged neurons or necrotic brain tissue. PMID- 11433428 TI - Multiple caspases are involved in beta-amyloid-induced neuronal apoptosis. AB - beta-amyloid peptide (Abeta) has been implicated in the pathogenesis of Alzheimer disease and has been reported to induce apoptotic death in cell culture. Cysteine proteases, a family of enzymes known as caspases, mediate cell death in many models of apoptosis. Multiple caspases have been implicated in Abeta toxicity; these reports are conflicting. We show that treatment of cerebellar granule cells (CGC) with Abeta25-35 causes apoptosis associated with increased activity of caspases-2, -3 and -6. Selective inhibition of each of these three caspases provides significant protection against Abeta-mediated apoptosis. In contrast, no change in caspase-1 activity was seen after Abeta25-35 application, nor was inhibition of caspase-1 neuroprotective. Similar to CGC, cortical neuronal cultures treated with Abeta25-35 demonstrate increased caspase-3 activity but not caspase-1 activity. Furthermore, significant neuroprotection is elicited by selective inhibition of caspase-3 in cortical neurons administered Abeta25-35, whereas selective caspase-1 inhibition has no effect. Taken together, these findings indicate that multiple executioner caspases may be involved in neuronal apoptosis induced by Abeta. PMID- 11433429 TI - Changes in gene expression following induction of ischemic tolerance in rat brain: detection and verification. AB - Tolerance against ischemic insults can be elicited in the CA1 region of rat hippocampus by inducing a short ischemic period 2-3 days prior to the ischemic insult. To detect genes whose expression changes following induction of ischemic tolerance (IT), we applied a differential display technique called restriction fragment differential display-PCR (RFDD-PCR). RFDD-PCR displays the coding region of mRNA and allows detection of differentially expressed mRNA. Double-stranded cDNA generated using a T25V primer is digested by the endonuclease TaqI, and adapters are ligated onto the cDNA fragments. When amplifying the adapter containing cDNA fragments under high-stringency conditions, reproducible PCR profiles are obtained. By comparing these profiles from naive and ischemia tolerant rat brains statistically, significant expression changes of 20 fragments were identified. To verify the observed changes, quantitative PCR and in situ hybridization were performed for three fragments representing proteins with quite different functions (GluR2-flop, SC1, and p68 RNA helicase). Quantitative PCR displayed the same degree of regulation as RFDD-PCR, but in situ hybridization did not display any regulation. As the applied PCR-based techniques detect only polyadenylated mRNA, whereas in situ hybridization detects both nonadenylated and adenylated mRNA, changes in the polyadenylation state of the mRNA, rather than inconsistent changes in the total amount of mRNA, probably explain this discrepancy. Thus, our results show that the expression of genes hitherto not related to IT changes with the induction of IT and that the degree of regulation displayed by RFDD-PCR can be verified by quantitative PCR. PMID- 11433430 TI - Interferon-beta treatment of experimental autoimmune encephalomyelitis leads to rapid nonapoptotic termination of T cell infiltration. AB - We investigated the possible mechanisms how interferon (IFN)-beta may control T cell infiltration in the CNS in experimental autoimmune encephalomyelitis (EAE). Adoptive transfer (AT) EAE was induced in groups of six female Lewis rats. Animals were treated with 3 x 10(5) units of recombinant rat IFN-beta s.c. once at 18 hr, or with 10 mg/kg methylprednisolone (MP) i.v. twice at 18 and 6 hr prior to dissection, or with a combination of both. T cell apoptosis was detected by immunohistochemistry on paraffin sections of spinal cord, using morphological criteria and TUNEL staining. Double labeling of immune cells was done for tumor necrosis factor (TNF)-alpha and metalloproteinase (MMP) 2. Disruption of the blood-brain barrier (BBB) was visualized by staining for albumin. In severe EAE, an increase of T cell apoptosis was seen after IFN-beta alone (all data presented as mean +/- SD: 24.5% +/- 2.2%, P < 0.05, vs. 19.4% +/- 3.1% in controls), and in combination with MP (29.4% +/- 7.3%, P < 0.05 vs. controls). Only the combination therapy decreased T cell infiltration (53.9 +/- 17.7 cells/mm(2), P < 0.05, vs. 99.5 +/- 35.2 cells/mm2 in controls). In moderate EAE, the rate of T cell apoptosis was slightly increased after IFN-beta (21.2% +/- 5.2% vs. 17.4% +/- 5.0% in controls), whereas MP alone (25.5% +/- 3.5%, P < 0.01 vs. controls) and the combination therapy (22.4% +/- 4.8%, P < 0.05 vs. controls) had a clear augmenting effect. IFN-beta tended to decrease T cell infiltration (46.1 +/- 12.7 cells/mm2) compared to controls (59.2 +/- 18.5 cells/mm2). The rate of TNF-alpha expressing T cells was significantly decreased by IFN-beta and in combination with MP. Also, TNF-alpha expression in macrophages was significantly reduced by IFN-beta and by the combination therapy. The rate of MMP2-expressing macrophages was lower after IFN-beta but clearly decreased only in combination with MP. BBB disruption was ameliorated after IFN-beta but significantly only in combination with MP. Our study indicates that IFN-beta affects the immunopathological process in EAE in several ways, but apoptosis appears as a minor component. In view of treatment of MS relapses, the synergistic effects in this study corroborate the use of a combination therapy with high-dose MP and IFN-beta. PMID- 11433431 TI - Synucleins in ocular tissues. AB - Synucleins are small proteins associated with neurodegenerative diseases and some forms of cancer. Most studies of this group of proteins have been directed to the elucidation of their role in the brain and their connection to the formation of depositions in brain tissues. Here we describe the localization of different types of synucleins in ocular tissues. By Western blot analysis, all members of the synuclein family are found in the retina and optic nerve, where their relative ratio varies. The data on immunohistochemical staining show that different members of the synuclein family have different localizations in ocular tissues. Alpha-synucleins and beta-synucleins are present predominantly in the inner plexiform layer, whereas gamma-synuclein is in the nerve fiber layer. In transgenic mice overexpressing alpha-synuclein, a different pattern of localization depending on the promoter used for the expression was observed. In Alzheimer's disease patients, immunohistochemical staining for gamma-synuclein revealed the loss of immunoreactivity in the nerve fiber layer and the nerve fiber layer and the appearance of immunopositive cells in or near the outer nuclear layer. We conclude that, in mature eyes, synucleins are present predominantly in the retina and optic nerve, and the immunoreactivity of gamma synuclein changes specifically in the retina of Alzheimer's disease patients. In transgenic mice overexpressing alpha-synuclein, immunopositive deposits in the optic nerve and accumulation of immunoreactivity in specific retinal cells were found. PMID- 11433432 TI - Cysteine dioxygenase: regional localisation of protein and mRNA in rat brain. AB - Cysteine dioxygenase (CDO) converts cysteine to cysteinesulphinic acid and is the rate-limiting step in sulphate production. Most studies have centred upon the hepatic form of the enzyme, but several studies have investigated brain CDO using activity assays and western blotting. The aim of this study was to investigate the expression of CDO in the rat brain using a combination of immunohistochemistry and in situ hybridisation. Affinity-purified anti-R and anti H CDO antibodies were immunoprecipitated using rat brain homogenate to determine whether the antibodies could remove enzyme activity. Immunohistochemistry and in situ hybridisation were then used to determine the cellular and regional expression of both CDO protein and mRNA. Immunoprecipitation of rat brain homogenate removed up to 98% and 70% (anti-R and anti-H, respectively) of enzyme activity. Nonimmune sheep serum had no effect upon enzyme activity. CDO protein and mRNA was localised solely to the neurones of the brain, including the pyramidal cells of the hippocampus and the Purkinje cells of the cerebellum. Regional localisation varied, with high levels of expression in the hippocampus, the dentate gyrus, the outer cortices of the brain, and the substantia nigra. The relative expression of CDO activity and protein in these regions is most probably a result of the relative abundance of neurones in these regions. CDO expression in the brain may have several possibilities functions, the most likely being the prevention of free radical production by the autoxidation of cysteine and dopamine. PMID- 11433435 TI - Click Chemistry: Diverse Chemical Function from a Few Good Reactions. AB - Examination of nature's favorite molecules reveals a striking preference for making carbon-heteroatom bonds over carbon-carbon bonds-surely no surprise given that carbon dioxide is nature's starting material and that most reactions are performed in water. Nucleic acids, proteins, and polysaccharides are condensation polymers of small subunits stitched together by carbon-heteroatom bonds. Even the 35 or so building blocks from which these crucial molecules are made each contain, at most, six contiguous C-C bonds, except for the three aromatic amino acids. Taking our cue from nature's approach, we address here the development of a set of powerful, highly reliable, and selective reactions for the rapid synthesis of useful new compounds and combinatorial libraries through heteroatom links (C-X-C), an approach we call "click chemistry". Click chemistry is at once defined, enabled, and constrained by a handful of nearly perfect "spring-loaded" reactions. The stringent criteria for a process to earn click chemistry status are described along with examples of the molecular frameworks that are easily made using this spartan, but powerful, synthetic strategy. PMID- 11433436 TI - Strategies for the Construction of Supramolecular Compounds through Coordination Chemistry. AB - Synthetic organic chemists enjoy the luxury of having a large collection of reliable reactions at their disposal for preparing small molecules, mesoscopic structures, and polymers. Coordination chemists, on the other hand, are faced with the fact that transition metal chemistry, when normalized for the number of transition metals, has relatively few high-yielding reactions, when compared to the chemistry of carbon, for preparing even small molecule structures. This lack of control is manifested, in large part, in the weak metal-ligand interactions found in coordination complexes as compared with the strong covalent bonds in organic compounds. Weak bonding often translates into many reaction pathways that are not substantially different from an energetic point of view, and therefore, results in poor selectivity. As a result, many coordination chemists in recent years have come to the realization that it may be easier and more productive to develop straightforward and reliable routes to mesoscopic supramolecular structures by capitalizing on the modest collection of high-yielding reactions in coordination chemistry, the directional bonding afforded by metal centers, and strategies aimed at taking advantage of the weak metal bonds found in coordination complexes. Three emerging synthetic strategies, the symmetry interaction, directional-bonding, and weak-link synthetic approaches, all use metal centers as structural building blocks to rationally assemble molecular components into supramolecular metallocyclophanes. These three approaches are discussed herein, and the fundamental principles underlying each as well as their capabilities are compared and contrasted. PMID- 11433437 TI - In Search of Practical Esterification. PMID- 11433438 TI - Recent Advances in High-Resolution Solid-State NMR Spectroscopy We thank the Massachusetts Institute for Technology, the Karl Winnacker Foundation, and the NIH under the NCRR program (RR00995) for financial support. We thank Elke Duchardt for helping us prepare the figures. PMID- 11433439 TI - Turning a Transition State into a Minimum-The Nature of the Bonding in Diplumbylene Compounds RPbPbR (R=H, Ar) Theoretical Studies of Inorganic Compounds, Part 15. This work was supported by the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie. We thank Prof. M. Weidenbruch for helpful suggestions. Part 14: Y. Chen, M. Hartmann, G. Frenking, Eur. J. Inorg. Chem., in press. PMID- 11433440 TI - A Novel Route to Fulvene Complexes of Titanium-Diastereoselective Complexation of Pentafulvenes to Cyclopentadienyltitanium Fragments Fulvene Complexes of Early Transition Metals, Part 1. This work was supported by the Bayer AG, Leverkusen (J.H.), the Bundesministerium fur Bildung und Forschung (J.S.,03C0276G/4), the Fonds der Chemischen Industrie, and the Karl-Ziegler-Stiftung of the GDCh. PMID- 11433441 TI - Fabrication of Micro-Chain Mail by Simultaneous, Patterned Electrodeposition on a Plane and Multiple Cylinders G.M.W. and H.W. gratefully acknowledge support of this research from the Defense Advanced Research Projects Agency and the National Science Foundation (ECS-9729405). S.W. thanks the NSERC and FCAR for funding. PMID- 11433442 TI - Color-Tuned Electroluminescence from Columnar Liquid Crystalline Alkyl Arenecarboxylates We are grateful to Nguyen Huu Tinh (Bordeaux), Joachim Wendorff and Jorg Kopitzke (Marburg), and Reinhold Oesterhaus (Paderborn) for their help and advice, and to Bayer AG for a sample of Baytron P. PMID- 11433443 TI - Synthesis of Apoptolidinone This work was supported by the Fonds der Chemischen Industrie, Schering AG, and the Schering Forschungsgesellschaft. We thank Prof. Dr. U. Eder (Schering AG, Berlin) for stimulating and helpful discussions. PMID- 11433444 TI - Carbon Nanofilaments in Heterogeneous Catalysis: An Industrial Application for New Carbon Materials? We thank the TIMCAL AG (Switzerland) for the generous provision of the graphite. PMID- 11433445 TI - Linear Polymeric C(70)(2-) Ions This work was supported by the Fonds der Chemischen Industrie. PMID- 11433446 TI - Orthogonal Photolysis of Protecting Groups Financial support from the Swiss National Science Foundation (grant no.: 21-57044.99) and the Fonds Frederic Firmenich et Philippe Chuit is gratefully acknowledged. PMID- 11433447 TI - Rate Enhancement and Enantioselectivity of the Jacobsen-Katsuki Epoxidation: The Significance of the Sixth Coordination Site This work was made possible through generous allocation of computer resources by the OIT at Notre Dame, the National Computational Science Alliance and the Competence Center for Computational Chemistry at ETH Zurich. PMID- 11433448 TI - Cyclohexylether delta-Amino Acids: New Leads for Selectivity Filters in Ion Channels Financial support by the Fonds der Chemischen Industrie (FCI), the Volkswagen Foundation, the Pinguin Foundation, and Schering AG is gratefully acknowledged. H.-D.A. thanks the FCI for a PhD fellowship. We thank Dr. B. Ziemer (Humboldt-Universitat zu Berlin) for the X-ray structure analysis and Dr. P. Franke (Freie Universitat Berlin) for MALDI-TOF mass spectra. PMID- 11433449 TI - A 2',4'-Bridged Nucleic Acid Containing 2-Pyridone as a Nucleobase: Efficient Recognition of a C small middle dotG Interruption by Triplex Formation with a Pyrimidine Motif Part of this work was supported by a Grant-in-Aid for Scientific Research (B) (No. 12557201) from the Japan Society for the Promotion of Science. PMID- 11433450 TI - A General Solution to the Modular Synthesis of Polyketide Building Blocks by Kanemasa Hydroxy-Directed Nitrile Oxide Cycloadditions This work was supported by the ETH-Zurich, Hoffmann-LaRoche, Merck, and Novartis. J.W.B. thanks the National Science Foundation (USA) for a predoctoral fellowship. We are grateful to Boehringer-Ingelheim for a generous gift of (R)-3-buten-2-ol. PMID- 11433451 TI - Alkylative Preparation of alpha-Silylalkylmagnesium from R(3)SiCHBr(2) Using a Magnesate Reagent This work was supported by a Grant-in-Aid for Scientific Research (Nos. 09450341 and 10208208) from the Ministry of Education, Culture, Sports, Science, and Technology. A.I. acknowledges the JSPS Research Fellowship for Young Scientists for financial support. PMID- 11433452 TI - Fluorinated Bis(enyl) Ligands by Metal-Induced Dimerization of Fluorinated Allenes This work was supported by the Fonds der Chemischen Industrie and the Deutsche Forschungsgemeinschaft. We are indebted to the Hoechst AG (Frankfurt/Main) and DUPONT (Wilmington, DE) for some starting materials. We thank Dr. Dieter Zobel for the collection of X-ray data at 20 K. PMID- 11433453 TI - Classification of Terpenoids according to the Methylerythritolphosphate or the Mevalonate Pathway with Natural (12)C/(13)C Isotope Ratios: Dynamic Allocation of Resources in Induced Plants We thank Dr. H. Bischoff (Bayer AG, Wuppertal) for cerivastatin and Steffen Ruhlow for his committed technical assistance. PMID- 11433454 TI - New Transmembrane Polyene Bolaamphiphiles as Fluorescent Probes in Lipid Bilayers Thanks are given to Prof. E. Gratton and Dr. L. A. Bagatolli, University of Illinois at Urbana-Champaign, USA, for the experiments on the orientation of the probes in POPC vesicles, and to P. P. Garcia Alvarez for his help in the synthesis. This work was financed by the Spanish D.G.I. (Projects PB96-852 and BQU2000/1500). E.Q. acknowledges a predoctoral fellowship from the same source. PMID- 11433455 TI - Nanoscale Push-Push Dihydrophenanthrene Derivatives as Novel Fluorophores for Two Photon-Excited Fluorescence This work was supported by the Delegation Generale pour l'Armement (DGA), the Institut Curie, and the Centre National de la Recherche Scientifique (CNRS). L.V. and L.M. received fellowships from the DGA and CNRS, respectively. PMID- 11433456 TI - Remote Stereocontrol in Carbonyl Additions Promoted by Vinylstannanes We Thank the M.E.C. of Spain (PB96/0357) and the Junta de Castilla y Leon (VA43/98) for financial support. We are very much indebted to Dr. Santiago Garcia-Granda, Oviedo (Spain) for X-ray crystallographic assistance. PMID- 11433457 TI - Detection of TNT and Picric Acid on Surfaces and in Seawater by Using Photoluminescent Polysiloles This work was supported through DARPA's Tactical Sensors Program through a Space and Naval Warfare Systems Center Contract (N66001 98-C-8514) and the National Science Foundation, Division of Material Research (Grant No. DMR 9900034). The technical point of contact for this DARPA program is Dr. Edward Carapezza. PMID- 11433458 TI - 3-Rhoda-1,2-dioxolanes through Dioxygenation of a Rhodium-Ethene Complex by Air This work was supported by the Netherlands Organization for Scientific Research (NWO-CW). We thank Johnson Matthey Ltd. for the generous loan of RhCl(3) small middle dot3H(2)O. S. Thewissen and P. M. van Galen are gratefully acknowledged for the ESI-MS measurements and P. P. J. Schlebos for NMR measurements. PMID- 11433459 TI - A Striking, Multifaceted, Decalithium Aggregate with Carbanion, Organoamide, and Alkoxide Functionalities This work was supported by the Australian Research Council, an ARC QE11 Research Fellowship to P.C.A and an Australian Postgraduate Award to N.M.S. PMID- 11433460 TI - Self-Assembly of Nanometer-Scale Secondary Building Units into an Undulating Two Dimensional Network with Two Types of Hydrophobic Cavity M.J.Z. gratefully acknowledges the financial support of the NSF (DMR 0101641). PMID- 11433461 TI - Polygons and Faceted Polyhedra and Nanoporous Networks M.J.Z. gratefully acknowledges the financial support of the NSF (DMR 0101641). PMID- 11433462 TI - A Rationally Designed NH(4)(+) Receptor Based on Cation-pi Interaction and Hydrogen Bonding We gratefully acknowledge the Korean Ministry of Science and Technology (Creative Research Initiative Program) for support of this work, and the Korean Ministry of Education (Brain Korea 21 program) for graduate studentships to J.H. and W.S.J. M.K. is grateful to POSTECH for an Undergraduate Research Fellowship. We also thank Professor P. K. Bharadwaj for helpful discussions. PMID- 11433463 TI - Macrocycles within Macrocycles: Cyclen, Cyclam, and Their Transition Metal Complexes Encapsulated in Cucurbit PMID- 11433464 TI - A Perfluorinated Nanosphere: Synthesis and Structure of Perfluoro-deca-B-methyl para-carborane This research was funded by the National Science Foundation (USA) (NSF CHE 9314037) and (NSF CHE 9972888). We gratefully thank graduate student Theodore W. Bitner and Prof. Jeffrey I. Zink, UCLA, for the acquisition of the Raman spectra. PMID- 11433465 TI - Modular Self-Assembly of a Microporous Solid Based upon Mercuracarborand-4 and a New Bonding Motif This work was supported by the National Science Foundation (CHE 9730006 and equipment grant CHE-9871332). PMID- 11433467 TI - Titanocene-Catalyzed Coupling of Amides in the Presence of Organosilanes To Form Vicinal Diamines This work was supported by the Natural Sciences and Engineering Research Council (Canada) and the Fonds FCAR du Quebec. PMID- 11433466 TI - Optically Tuning the Rate of Stoichiometry Changes: Surface-Controlled Oxygen Incorporation into Oxides under UV Irradiation. PMID- 11433468 TI - High-Valent Manganese Corroles and the First Perhalogenated Metallocorrole Catalyst This research (No. 368/00) was supported by the Israel Science Foundation (Z.G.), the US National Science Foundation (H.B.G.), and the Fund for the Promotion of Research at the Technion (Z.G.). PMID- 11433469 TI - Self-Assembly of Uniform Spherical Aggregates of Magnetic Nanoparticles through pi-pi Interactions The authors thank the Special Research Foundation of the National Nature Science Foundation of China (29992530), State Key Project Fundamental Research (G1999064504) for continuing financial support. PMID- 11433470 TI - Dehydrogenative Dimerization of Tin Hydrides Catalyzed by Ruthenium-Allenylidene Complexes We thank The Skaggs Institute for Chemical Biology at The Scripps Research Institute for support of this work; S.M.M. is a Skaggs Postdoctoral Fellow. We are also grateful to Dr. Michal Sabat (University of Virginia) for X ray crystallography and Prof. David Goodin (The Scripps Research Institute) for the EPR experiment described in ref. 14. PMID- 11433471 TI - Novel Templating Effect in the Macrocyclization of Functionalized Diynes by Zirconocene Coupling This work was supported by the Director, Office of Basic Energy Sciences, Chemical Sciences Division, of the U.S. Department of Energy under Contract No. DE-AC03-76SF00098. The Center for New Directions in Organic Synthesis is supported by Bristol-Myers Squibb as Sponsoring Member. We also thank Dr. F. J. Hollander of the U.C. Berkeley X-ray diffraction facility (CHEXRAY) for help with determination of the X-ray structures. PMID- 11433472 TI - Rapid Access to Complex Molecular Architectures via o-Azaquinones We thank Dr. D. H. Huang, G. Siuzdak, and Dr. R. Chadha for NMR spectroscopic, mass spectrometric, and X-ray crystallographic assistance, respectively. This work was supported financially by The Skaggs Institute for Chemical Biology, the National Institutes of Health (USA), a predoctoral fellowship from the National Science Foundation (P.S.B), and grants from ArrayBiopharma, Pfizer, Glaxo, Merck, Schering Plough, Hoffmann-La Roche, Boehringer Ingelheim, DuPont, and Abbott Laboratories. PMID- 11433473 TI - Separation of Spliceosome Assembly from Catalysis with Caged pre-mRNA Substrates A.M.M. acknowledges support from the Natural Sciences and Engineering Research Council of Canada and the Alberta Heritage Foundation for Medical Research. PMID- 11433474 TI - Base-Assisted Formation of Organozirconium Oxides with the PMID- 11433475 TI - A Luminescent Gold Ring That Flips Like Cyclohexane This work was supported by the National University of Singapore. We thank Dr. Leong Weng Kee and Dr. Richard Wong for helpful discussion and Dr. Jagadese J. Vittal and Ms. G. K. Tan for determining the X-ray crystal structures. PMID- 11433476 TI - Self-Assembling Cyclic Peptide Cylinders as Nuclei for Crystal Engineering This work was supported in part by the Office of Naval Research and the National Institutes of Health (GM 52190). We thank R.K. Chadja for crystallographic analysis and N. Kresge for assistance with computer graphics representations. PMID- 11433477 TI - A Large 24-Membered-Ring Germanate Zeolite-Type Open-Framework Structure with Three-Dimensional Intersecting Channels This work was supported by the NSF of China (Grant No. 29873012 and 29925309), National Education Ministry, State Key Basic Research Program of PRC (G2000048001) Shanghai Science Foundation, and the Key Laboratory of Inorganic Hydrothermal Synthesis, Jilin University. We thank Prof. S. Feng and Dr. Z. Shi for invaluable discussions. PMID- 11433478 TI - Formation of Aryl- and Benzylboronate Esters by Rhodium-Catalyzed C-H Bond Functionalization with Pinacolborane The EPSRC is gratefully acknowledged for financial support (T.B.M. and J.A.K.H.) and for a Senior Fellowship (J.A.K.H.). Dr. Shimada thanks the Science and Technology Agency of Japan for a fellowship, and Dr. M. Tanaka for granting sabbatical leave, making possible his stay in Durham. We thank Prof. M. Srebnik for advice on the purification of HBpin. PMID- 11433479 TI - Stable, Monomeric Imides of Aluminum and Gallium: Synthesis and Characterization of PMID- 11433480 TI - The Novel Open-Framework Vanadium Silicates K(2)(VO)(Si(4)O(10)) small middle dotH(2)O (VSH-1) and Cs(2)(VO)(Si(6)O(14)) small middle dot3 H(2)O (VSH-2) We thank the National Science Foundation (DMR-9805881) and the Robert A. Welch Foundation for support. This work made use of MRSEC/TCSUH Shared Experimental Facilities supported by the National Science Foundation under Award Number DMR 9632667 and the Texas Center for Superconductivity at the University of Houston. PMID- 11433482 TI - Protein Crystals as Novel Catalytic Materials. AB - In this era of molecular biology, protein crystallization is often considered to be a necessary first step in obtaining structural information through X-ray diffraction analysis. In a different light, protein crystals can also be thought of as materials, whose chemical and physical properties make them broadly attractive and useful across a larger spectrum of disciplines. The full potential of these protein crystalline materials has been severely restricted in practice, however, both by their inherent fragility, and by strongly held skepticism concerning their routine and predictable growth, formulation, and practical application. Fortunately, these problems have turned out to be solvable. A systematic exploration of the biophysics and biochemistry of protein crystallization has shown that one can dependably create new protein crystalline materials more or less at will. In turn, these crystals can be readily strengthened, both chemically and mechanically, to make them suitable for practical commercialization. Today, these novel materials are used as industrial catalysts on a commercial scale, in bioremediation and "green chemistry" applications, and in enantioselective chromatography of pharmaceuticals and fine chemicals. In the near future, their utility will expand, to include the purification of protein drugs, formulation of direct protein therapeutics, and development of adjuvant-less vaccines. PMID- 11433483 TI - Programming Organic Molecules: Design and Management of Organic Syntheses through Free-Radical Cascade Processes. AB - Cascade, domino, or tandem processes, that link together two or more transformations in one pot, are increasing in popularity because they lead to improvements in synthetic efficiency and decreases in environmental impact. Not only do these cascades contain choice mechanistic gems but they also deliver compact and elegant syntheses of complex natural products. Longer cascades require more functional groups precisely configured within carefully designed initial molecular architectures. Such "purposeful" molecules can be thought of as chemical algorithms.This article surveys the phenomenal range of unimolecular free-radical cascades. A convenient system for classifying free-radical cascades is described that is useful for evaluating and comparing cascades and aids the design of synthetic routes to polycyclic structures.Double cyclization cascades lead to cyclopentylcyclopentane or bicyclo[3.3.0]octane derivatives. Precursors that contain a ring as a template have been used to control stereochemistry in syntheses of triquinanes and many related compounds. Of the cascades containing ring-cleavage steps, the most useful are the ring expansions which have opened up new synthetic routes to medium ring polycycles.The key design features of three stage unimolecular free-radical cascades that yielded steroid structures, are linear arrays of radical acceptor units associated with methyl groups distributed every fifth C-atom in the precursor polyenes. Ring cleavage is the reverse of cyclization. In special, symmetrical structures, therefore, this led to sequences that were reversible, thus launching endlessly repeating cascades supported by delightfully fluxional structures. The science of "programming" organic molecules to achieve particular target structures is maturing rapidly. Coordination and classification of the welter of information in this area is intended to facilitate design and hence to extend the range and complexity of attainable structures. PMID- 11433484 TI - TNA as a Potential Alternative to Natural Nucleic Acids. PMID- 11433485 TI - Fascinating Natural and Artificial Cyclopropane Architectures. PMID- 11433486 TI - Beneficial Effect of ortho-Methoxy Groups in the Asymmetric Ring Opening of meso Epoxides with Silicon Tetrachloride Catalyzed by Chiral ortho Methoxyphenyldiazaphosphonamide Lewis Bases Response to the Communication by G. Buono et al. We are grateful to the National Science Foundation (CHE-9803124) for generous financial support. Cory M. Stiff is thanked for preliminary experiments. PMID- 11433487 TI - Gold Glyconanoparticles as Water-Soluble Polyvalent Models To Study Carbohydrate Interactions This work was supported by the DGICYT (PB96-0820), J.M.F. thanks the MEC for a predoctoral fellowship. A.G.B. thanks CSIC for financial support. We thank Prof. Martin-Lomas for his scientific and financial support. PMID- 11433488 TI - Electronic Transduction of Polymerase or Reverse Transcriptase Induced Replication Processes on Surfaces: Highly Sensitive and Specific Detection of Viral Genomes Parts of this research are supported by the Israel Ministry of Science as an Infrastructure Project in Biomicroelectronics and as an Israel Japan cooperation. M.K. acknowledges the support of the American Foundation for Aids Research, AmfAR (grant No. 02730-28-RG). PMID- 11433489 TI - Cesium- and Rubidium-Selective Redox-Active Bis(calix PMID- 11433490 TI - Cyclic Hexamer with a Cubic Cavity: Crystal Structure of PMID- 11433491 TI - Active Site Design in a Chemzyme: Development of a Highly Asymmetric and Remarkably Temperature-Independent Catalyst for the Imino Aldol Reaction This work was supported by a grant from UOP Company. We would like to thank Thomas Malloy, Timothy Brandvold and Beth McCulloch for stimulating discussions. PMID- 11433492 TI - Cu(2+) Inhibits the Aggregation of Amyloid beta-Peptide(1-42) in vitro We thank JEOL for the AFM measurement. This work was supported in part by Grants-in-Aid from the Japanese Ministry of Education, Science, Sports, and Culture, and a Grant from "Research for the Future" Program of the Japan Society for the Promotion of Science to N.S. PMID- 11433493 TI - Pure Polymorph C of Zeolite Beta Synthesized by Using Framework Isomorphous Substitution as a Structure-Directing Mechanism We thank the Spanish CICYT for financial support (Project MAT2000-1392). One of authors (J.R.) thanks the "Direccion General de Investigacion" of the MCYT for financial help (PB98-0483). M.T.N thanks to the Fundacion Jose y Ana Royo for a postdoctoral grant. PMID- 11433494 TI - Hydrophobic Pockets in a Nonpolymeric Aqueous Gel: Observation of such a Gelation Process by Color Change We thank the Jawaharlal Nehru Center for Advanced Scientific Research (Bangalore), and Mitokor, Inc. (San Diego) for supporting this work through unrestricted research grants. PMID- 11433496 TI - Bi(4)Te(4)(4+)-A Cube-Shaped, Polycationic Main Group Element Cluster This work was supported by the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie. PMID- 11433495 TI - Two Isomers of C(60)F(48): An Indented Fullerene This work was supported by the Russian Program "Fullerenes and Atomic Clusters" and RFBR (grant 99-15-96044m). We are grateful to Dr. D. N. Laikov (Organic Chemistry Institute, Zurich University) and the High-Performance Computing Center of the Chemistry Department of the Moscow State University for their assistance in performing calculations. PMID- 11433497 TI - Hybrid Molecular Materials Based on Covalently Linked Inorganic Polyoxometalates and Organic Conjugated Systems We thank the University of Missouri-Kansas City and Research Corporation for supporting this work. PMID- 11433498 TI - Chemoenzymatic-Chemical Synthesis of a (2-3)-Sialyl T Threonine Building Block and Its Application to the Synthesis of the N-Terminal Sequence of Leukemia Associated Leukosialin (CD 43) This work was supported by the Deutsche Forschungsgemeinschaft, the Stiftung Rheinland Pfalz fur Innovation, and the Fonds der Chemischen Industrie. PMID- 11433499 TI - Matrix Isolation of Perfluorinated p-Benzyne W.S., H.H.W., and A.B. thank the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie, W.T.B. and D.A.H. the National Science Foundation for financial support. The systematic name for perfluorinated p-benzyne is 1,4-didehydro-2,3,5,6-tetrafluorobenzene. PMID- 11433500 TI - Terminal Borylene Complexes as a Source for the Borylene B-N(SiMe(3))(2): Alternative Synthesis and Structure of PMID- 11433501 TI - The PMID- 11433502 TI - Inversion of Enantioselectivity during the Platinum-Catalyzed Hydrogenation of an Activated Ketone This work was supported by the Swiss National Science Foundation. We thank F. Bangerter and D. Sutter for NMR measurements. PMID- 11433503 TI - The Mechanism of the PMID- 11433504 TI - E(SiMe(3))(4)(+) Ions (E=P, As): Persilylated Phosphonium and Arsonium Ions This work was supported by the Deutsche Forschungsgemeinschaft, the Fonds der Chemischen Industrie, and the Ministerium fur Schule, Wissenschaft und Forschung des Landes Nordrhein-Westfalen. PMID- 11433505 TI - Diastereocontrol in the Synthesis of Models of Rings C and D of Phorbol: Directing Effect of an Ether Substituent on Lithium Carbenoid Mediated Cyclopropanation We thank Mr. A. W. Jahans for technical assistance. A.J.M.-S. thanks the Secretaria de Estado de Universidades, Investigacion y Desarrollo (SEUID; Spanish funding body) for a postdoctoral fellowship. R.S. thanks Astra Zeneca for a research studentship. PMID- 11433506 TI - Constructing Tricyclic Compounds Containing a Seven-Membered Ring by Ruthenium Catalyzed Intramolecular PMID- 11433507 TI - Correlating Electron Transport and Molecular Structure in Organic Thin Films This work was supported by the Office of Naval Research, the Defense Advanced Research Project Agency, and the National Science Foundation ECS-97294053. R.E.H. thanks the National Institutes of Health for a postdoctoral fellowship, and R.H. thanks the Deutsche Forschungsgemeinschaft and the BASF fellowship program for financial support. We thank Andreas Terfort for the synthesis of aromatic thiols. Collaboration between Caracas and Evanston is supported by NSF Conicit. PMID- 11433508 TI - Quantitative Studies of Binding between Synthetic Galactosyl Ceramide Analogues and HIV-1 Gp120 at Planar Membrane Surfaces This research was supported by the NIH (AI40359-02) and the NSF (CHE-9726132). Scientific discussions with Prof. Dr. J. C. Conboy (University of Utah) and Prof. Dr. J. Gervay-Hague (University of Arizona) are gratefully acknowledged. PMID- 11433509 TI - Metal Complexes of an N-Confused Calix PMID- 11433510 TI - Fragmentation of Carbohydrate Anomeric Alkoxy Radicals: A New Synthesis of Chiral 1-Halo-1-iodo Compounds This work was supported by the Investigation Programme no. PB96-1461 of the Direccion General de Investigacion Cientifica y Tecnica, Spain. C.C.G. and C.R.-F. thank the Ministerio de Educacion y Ciencia, Spain, and the Direccion General de Universidades e Investigacion del Gobierno de Canarias, respectively, for fellowships. PMID- 11433511 TI - A Schizophrenic Water-Soluble Diblock Copolymer We thank EPSRC for a post doctoral fellowship for S.Y.L. (GR/N17409). Dr. P. McKenna of Laporte Performance Chemicals, Hythe (UK), is thanked for the gift of the monohydroxy-capped PPO. The reviewers are thanked for their helpful and constructive comments. PMID- 11433512 TI - sigma-Bond Metathesis of Alkanes on a Silica-Supported Tantalum(V) Alkyl Alkylidene Complex: First Evidence for Alkane Cross-Metathesis We are grateful to Dr. L. Lefort (LCOMS) and Dr. B. Maunders (BP Chemicals) for fruitful discussions. We also wish to thank the C.N.R.S., C.P.E. Lyon, and BP Chemicals for financial support. PMID- 11433513 TI - 1,3-Diethynylallenes: New Modules for Three-Dimensional Acetylenic Scaffolding We thank the ETH Research Council and the Fonds der Chemischen Industrie for their support of this work. PMID- 11433514 TI - Directed Intermolecular Carbomagnesation across Vinylsilanes: 2-PyMe(2)Si Group as a Removable Directing Group This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture, Japan, and in part by the Mitsubishi Foundation. K.M. thanks the Japan Society for the Promotion of Science for Young Scientists. PMID- 11433515 TI - Pentacoordination of Silicon by Four Covalent Si-S Bonds and One Covalent Si-C Bond This work was supported by the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie. PMID- 11433516 TI - [Cu] PMID- 11433517 TI - A Superoxovanadium(V) Complex Linking the Peroxide and Dioxygen Chemistry of Vanadium This work was supported by the Deutsche Forschungsgemeinschaft. PMID- 11433518 TI - Self-Assembly and Manipulation of Crown Ether Phthalocyanines at the Gel-Graphite Interface This work was supported by the EU-TMR project SISITOMAS (project reference FMRX970099), the European Science Foundation through SMARTON, and the Volkswagen-Stiftung. PMID- 11433519 TI - [ReH] PMID- 11433520 TI - The Hg(3)(2+) Group as a Framework Unit in a Host-Guest Compound: PMID- 11433521 TI - Karyotypic similarity identified by multiplex-FISH relates four prostate adenocarcinoma cell lines: PC-3, PPC-1, ALVA-31, and ALVA-41. AB - Recently developed molecular cytogenetic techniques for karyotyping are providing new and important insights regarding the chromosomal changes that occur in solid tumors. We used multiplex-FISH to analyze four adenocarcinoma cell lines, PC-3, PPC-1, ALVA-31, and ALVA-41, in which the characterization of a large number of rearranged chromosomes was partially or substantially inconclusive by G-banding. Although the original descriptions of these lines depict them as distinct entities established from different patients, this study demonstrates that these four lines share numerous, highly rearranged chromosomes, strongly supporting the conclusion that they are derived from the same patient material. Our analysis indicates that PPC-1, ALVA-31, and ALVA-41 were derived from PC-3 through mechanisms involving clonal progression represented by sequential changes and clonal diversion represented by differing patterns of changes. Extensive cellular heterogeneity was detected in all four lines, and most rearrangements included segments derived from multiple chromosomes. Each line also showed a set of unique derivative chromosomes. However, a limited number of metaphase cells (approximately 10) was analyzed for each line, and numerous single-cell abnormalities were detected in all of them. Therefore, it is plausible that the number of clonal, shared, and/or unique rearrangements has been underestimated. These cell lines have been utilized as models for understanding the biology of prostate cancer and reportedly differ in their cell physiology. Rather than detracting from their value, a complete understanding of the interrelationships of these lines to one another may provide the opportunity to define the molecular changes that have led to their individual malignant phenotypes. PMID- 11433522 TI - Molecular-cytogenetic comparison of mucosa-associated marginal zone B-cell lymphoma and large B-cell lymphoma arising in the gastro-intestinal tract. AB - Extranodal B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) type may represent a model of lymphoma progression, because a small cell component frequently occurs in the large cell variants. We studied 52 extranodal B-cell lymphomas: 18 extranodal marginal zone B-cell lymphomas of MALT type (MZBL,MT), 7 MZBL,MT of the gastro-intestinal tract with a diffuse large B-cell component (giMZBLplusLBCL), and 27 diffuse large B-cell lymphomas of the gastro-intestinal tract without small cell component (giLBCL). Analytical techniques were comparative genomic hybridization (CGH) and fluorescence in situ hybridization (FISH). The translocation t(11;18) was found as the sole aberration in two MZBL,MT only. In contrast to this, t(11;18)-negative MZBL,MT were characterized by frequent gains on chromosome 3 and DNA amplifications on 2p13-p15. Furthermore, we found a clonal lymphoma progression from the small to the large cell component with accumulation of gains and losses of chromosomal material in the large cell component in giMZBLplusLBCL. Aberrations overlapping with MZBL,MT and giMZBLplusLBCL included losses on chromosome 13, amplifications of the REL proto-oncogene, or gains on chromosome 12. In addition, the large cell component revealed gains on 8q24, including amplifications of the MYC proto-oncogene, and losses on 2q. The giLBCL had frequent gains on chromosomes 12 and 9, as well as on 11q, and losses on 6q. We conclude that, based on the distinctive and partly overlapping patterns of genetic aberrations, MALT lymphomas can be divided into different genetic subgroups. PMID- 11433523 TI - Homozygous deletion in a neuroblastoma cell line defined by a high-density STS map spanning human chromosome band 1p36. AB - Recent molecular studies have shown a relatively high rate of loss of heterozygosity (LOH) in neuroblastoma (NB) as well as other types of tumors in human chromosome band 1p36. To identify candidate tumor suppressor genes in NB, we searched for homozygous deletions in NB cell lines with PCR according to a high-density sequence tagged site (STS)-content map spanning 1p35-36. Among 25 NB cell lines examined, only one cell line, NB-1, showed no signal with 27 STSs in a 480 kb region in 1p36.2. The sequence analysis has revealed that the defective region included seven known genes (E4, KIF1B, SCYA5, PGD, Cortistatin, DFF45, and PEX14), nine expressed sequence tags (ESTs), and two microsatellite markers. These genes are related to apoptosis, an ubiquitin-proteasome pathway, a neuronal microtubule-associated motor molecule, and components of a common translocation machinery. The region between the DFF45 and KIF1B genes was defined as homozygous deletion by Southern blotting. The search in LOH regions with high-density STSs may be useful for the isolation and identification of tumor suppressor genes in other tumors as well as NBs. PMID- 11433524 TI - Defining a common region of deletion at 13q21 in human cancers. AB - Previous molecular genetic analyses identified a region of deletion at 13q21 in a variety of human cancers, suggesting the existence of a tumor suppressor gene(s) at this locus. In our earlier study on prostate cancer, the region of deletion was confined to a 3.1 cM interval between D13S152 and D13S162. At present, however, no known gene located in this interval has been firmly implicated in cancer, and the region remains too large for gene identification. To fine-map the area of interest, we established a contig of bacterial artificial chromosome (BAC) clones, narrowed the region of deletion by loss of heterozygosity (LOH) and homozygosity-mapping-of-deletion (HOMOD) analyses in different types of cancers, and tested a candidate gene from the region for mutation and alteration of expression in prostate cancers. The contig consisted of 75 overlapping BAC clones. In addition to the generation of 47 new sequence-tagged-site (STS) markers from the ends of BAC inserts, 76 known STS and expressed sequence tag markers were mapped to the contig (25 kb per marker on average). The minimal region of deletion was further defined to be about 700 kb between markers D13S791 and D13S166 by LOH analysis of 42 cases of prostate cancer, and by HOMOD analysis of eight prostate cancer cell lines/xenografts and 49 cell lines from cancers of the breast, ovary, endometrium, and cervix, using 18 microsatellite markers encompassing the deletion region. A gene that is homologous to the WT1 tumor suppressor gene, AP-2rep (KLF12), was mapped in this region and was analyzed for its expression and genetic mutation. In addition to low levels of expression in both normal and neoplastic cells of the prostate, this gene did not have any mutations in a group of aggressive prostate cancers and cell lines/xenografts, as assessed by the methods of polymerase chain reaction-single strand conformational polymorphism analysis and direct sequencing. These studies suggest that a 700 kb interval at 13q21 harbors a tumor suppressor gene(s) that seems to be involved in multiple types of cancer, and that the AP-2rep gene is unlikely to be an important tumor suppressor gene in prostate cancer. The BAC contig and high resolution physical map of the defined region of deletion should facilitate the cloning of a tumor suppressor gene(s) at 13q21. PMID- 11433525 TI - Amplification of Mycn, Ddx1, Rrm2, and Odc1 in rat uterine endometrial carcinomas. AB - The BDII rat is genetically predisposed to estrogen-dependent endometrial adenocarcinoma and represents a valuable model for this type of tumor. Tumors arising in strain crosses involving the BDII rats had previously been screened for DNA copy number changes using comparative genome hybridization (CGH). It was found that extra copies of the proximal region of rat chromosome (RNO) 6 commonly could be detected in these tumors. Based on RH-mapping data and comparative mapping with mouse and human, seven cancer-related genes were predicted to be situated in RNO6q14-q16. Rat PACs were isolated for the N-myc proto-oncogene (Mycn), apolipoprotein B (Apob), the DEAD box gene 1 (Ddx1), ornithine decarboxylase 1 (Odc1), proopiomelanocortin (Pomc1), ribonucleotide reductase, M2 polypeptide (Rrm2), and syndecan 1 (Sdc1). The localization of the genes to the region was verified by FISH (fluorescence in situ hybridization) mapping, and the detailed order among them was determined by dual-color FISH. By Southern blot analysis, it was found that the Mycn locus was highly amplified in two out of 10 cell cultures derived from the tumors. In one of them (designated RUT30), the amplification level of Mycn was estimated at 140x. Two other genes were coamplified (Ddx1 and Rrm2) at much lower levels. Similarly, in another culture (designated RUT2), Mycn was amplified more than 40x, whereas three of the other genes (Ddx1, Rrm2, and Odc1) were coamplified at lower levels. Using FISH on metaphase chromosomes from the cell cultures analyzed, the amplified sequences were shown to be located in typical HSRs. With competitive RT-PCR, distinct overexpression of Mycn and Ddx1 could be demonstrated in both RUT2 and RUT30. In addition, Mycn was overexpressed in two other tumors not exhibiting Mycn amplification. Taken together, our results suggest that overexpression of Mycn plays an important role in the development of endometrial cancer in the BDII rat. In humans, Mycn amplification has been reported mainly from tumors of neuronal origin. To our knowledge, this is the first report of Mycn amplification and overexpression in hormone-dependent tumors. PMID- 11433526 TI - Methylation pattern of different regions of the MLH1 promoter and silencing of gene expression in hereditary and sporadic colorectal cancer. AB - Nonrandom, widespread promoter methylation of tumor suppressor genes is a common mechanism of gene inactivation during tumorigenesis. We examined the methylation status of two distinct regions of the MLH1 promoter (proximal and distal to the transcription start site) and the MLH1 gene expression by methylation-specific PCR and immunohistochemistry. A total of 72 colorectal tumors, both with (n = 51, 22 affected by hereditary nonpolyposis colorectal cancer, HNPCC, defined according to the international clinical criteria and 29 sporadic cases) and without microsatellite instability (MSI) (n = 21) were studied. Methylation was present in at least one of the two promoter regions in 86% of the sporadic MSI cases, in 33% of the cases lacking MSI, and in 23% of the HNPCC tumors. In the HNPCC cases with a known MLH1 mutation (n = 10) none of the two promoter regions was methylated. Hypermethylation in both MLH1 promoter regions was seen in 45% of the MSI sporadic cases vs. 5% of the MSI-negative cases and 0% of the HNPCC cases. The overall concordance between the two promoter regions regarding methylation status was good (P = 0.009), but no significant correlation between methylation and suppression of the MLH1 immunohistochemical expression was found. Our data confirm that mutation and hypermethylation are mutually exclusive mechanisms in inducing mismatch repair deficiency and support the hypothesis of methylation as a process evenly distributed along the different regions of the promoter. PMID- 11433527 TI - Clinical impact of molecular and cytogenetic findings in synovial sarcoma. AB - Synovial sarcoma is an aggressive soft-tissue tumor that accounts for up to 10% of soft-tissue sarcomas. Cytogenetically, synovial sarcoma is characterized by the t(X;18)(p11;q11), found in more than 95% of the tumors. This translocation results in rearrangements of the SYT gene in 18q11 and one of the SSX1, SSX2, or SSX4 genes in Xp11, creating a SYT/SSX1, SYT/SSX2, or SYT/SSX4 chimeric gene. It has been shown that patients with SYT/SSX1 fusion genes have a shorter metastasis free survival than do patients with SYT/SSX2. Previous studies have also suggested that clonal evolution may be associated with disease progression. In the present study, RT-PCR analysis showed that all 64 examined synovial sarcomas from 54 patients had SYT-SSX chimeric genes. SYT/SSX1 was found in 40 tumors from 33 patients, SYT/SSX2 in 23 tumors from 20 patients, and SYT/SSX4 in one case. Two patients had variant SYT/SSX2 transcripts, with 57 bp and 141 bp inserts, respectively, between the known SYT and SSX2 sequences. Patients with tumors with SYT/SSX1 fusions had a higher risk of developing metastases compared to those with SYT/SSX2 fusions (P = 0.01). The reciprocal transcripts SSX1/SYT and SSX2/SYT were detected using nested PCR in 11 of the 40 samples with SYT/SSX1 and 5 of the 23 samples with SYT/SSX2, respectively. Among 20 blood samples, SYT/SSX1 and SYT/SSX2 were detected in one sample each. The t(X;18), or variants thereof, was found cytogenetically in all patients but three. Among 32 primary tumors, the t(X;18) or a variant translocation was the sole anomaly in 10. In contrast, of the seven metastatic lesions that were investigated prior to radiotherapy, only one had a t(X;18) as the sole anomaly; all other tumors displayed complex karyotypes. Cytogenetic complexity in primary tumors was, however, not associated with the development of metastases. Tumors with SYT/SSX2 less often (4/12 vs. 7/15) showed complex karyotypes than did tumors with SYT/SSX1, but the difference was not significant. Combining cytogenetic complexity and transcript data, we found that the subgroup of patients with tumors showing simple karyotypes and SYT/SSX2 fusion had the best clinical outcome (2/8 patients developed metastases), and those with tumors showing complex karyotypes together with SYT/SSX1 fusion the worst (6/7 patients developed metastases). This corresponded to 5-year metastasis-free survival rates of 0.58 and 0.0, respectively (P = 0.02). PMID- 11433528 TI - Analysis of loss of heterozygosity in lymphoma and leukaemia arising in F1 hybrid mice locates a common region of chromosome 4 loss. AB - Previous studies have identified five lymphoma-related tumour suppressor gene regions on murine chromosome 4. Using detailed allelotype analysis on a range of lympho-haematopoietic tumour types arising in F1 hybrid mice, we now show a consistent pattern of loss of heterozygosity (LOH) which identifies a common region of loss delineated by microsatellites D4Mit21 and D4Mit53 on proximal chromosome 4. This critical segment corresponds to the thymic lymphoma tumour suppressor region 5 (TLSR5) identified in an earlier study. Tumours of this type have also been reported as showing allelic loss from the Trp53 and Ikaros regions on chromosome 11. In the present study, only a small fraction of tumours showed LOH in the Ikaros region, while a minority of lymphomas, but not acute myeloid leukaemias, showed allelic loss of the chromosome 11 segment encoding Trp53. These and other data indicate strongly that the genomic regions identified as showing recurrent LOH depend on the genetic background of the mice. Overall, the results indicate a key role for a tumour suppressor gene(s) encoded in an approximately 3 cM segment on proximal chromosome 4 and provide an experimental basis for the further investigation of the functional role of candidate genes which include Pax5 and Tgfbr1. PMID- 11433529 TI - Human LPP gene is fused to MLL in a secondary acute leukemia with a t(3;11) (q28;q23). AB - The mixed lineage leukemia, MLL, gene is frequently rearranged in patients with secondary leukemia following treatment with DNA topoisomerase II inhibitors. By FISH and Southern blot analyses we identified a rearrangement in the MLL gene due to a novel t(3;11)(q28;q23) chromosomal translocation in a patient who developed AML-M5 3 years after treatment for a follicular lymphoma. Through inverse PCR, the LPP (lipoma preferred partner) gene on 3q28 was identified as the MLL fusion partner. LPP contains substantial identity to the focal adhesion protein, zyxin, and is frequently fused to HMGIC in lipomas. The breakpoint occurred in intron 8 of MLL and LPP. Two in-frame MLL-LPP transcripts, which fuse MLL exon 8 to LPP exon 9, were detected by RT-PCR, although the smaller of these contained a deletion of 120 bp from the MLL sequence. The predicted MLL-LPP fusion protein includes the A/T hook motifs and methyltransferase domain of MLL joined to the two last LIM domains of LPP. A reciprocal LPP-MLL transcript, predicted to include the proline-rich and leucine zipper motifs, and the first LIM domain of LPP were also detected by RT-PCR. In summary, LPP is a newly identified MLL fusion partner in secondary leukemia resulting from topoisomerase inhibitors. The MLL-LPP and LPP-MLL predicted proteins contain many of the features present in other MLL rearrangements. PMID- 11433530 TI - Allelic loss on chromosome bands 13q11-q13 in esophageal squamous cell carcinoma. AB - Allelic loss on chromosome 13 occurs frequently in esophageal squamous cell carcinoma. However, studies of the two known tumor suppressor genes located on 13q, RB1 and BRCA2, have shown few mutations, suggesting that other genes are likely to be involved in the development of this tumor type. To identify a minimal deletion interval, we first analyzed 42 microsatellite markers spanning chromosome bands 13q11-q13 in 56 esophageal squamous cell carcinoma patients, including 34 with a family history of upper gastrointestinal cancer and 22 without a family history of cancer. Lifestyle risk factors and clinical/pathologic characteristics were also collected. Two commonly deleted regions were identified: one was located on band 13q12.11, between markers D13S787 and D13S221; the other was located on bands 13q12.3-q13.1 from markers D13S267 to D13S219. We observed higher allelic loss frequencies for eight of the microsatellite markers in those patients with a family history of upper gastrointestinal cancer compared to patients without such a history. This study suggests that one or more unidentified tumor suppressor genes are located on chromosome arm 13q that play a role in the development of esophageal squamous cell carcinoma. PMID- 11433531 TI - CDKN2A germline splicing mutation affecting both p16(ink4) and p14(arf) RNA processing in a melanoma/neurofibroma kindred. AB - The CDKN2A locus encodes two tumor suppressor proteins, p16(ink4) and p14(arf), through use of alternative first exons. CDKN2A mutations detected in melanoma families are usually missense or nonsense changes which mainly impair p16(ink4) function. Large genomic deletions spanning the entire locus have been observed in two pedigrees with melanomas and nervous tumors. We have detected a novel splice site mutation in a family with melanomas, neurofibromas, and multiple dysplastic nevi. Both alternative mRNAs produced by the mutant allele lacked shared sequences from exon 2, which encodes a substantial portion (>50%) of both p16(ink4) and p14(arf) proteins. The development of neurofibromas can be explained by cooperative effects of combined inactivation of p16(ink4) and p14(arf) or, alternatively, of p14(arf) alone. PMID- 11433532 TI - Perspectives on the use of electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry for short tandem repeat genotyping in the post-genome era. AB - The recent completion of the first rough draft of the human genome has provided fundamental information regarding our genetic make-up; however, the post-genome era will certainly require a host of new technologies to address complex biological questions. In particular, a rapid and accurate approach to characterize genetic markers, including short tandem repeats (STRs) and single nucleotide polymorphisms (SNPs) is demanded. STRs are the most informative of the two polymorphisms owing to their remarkable variability and even dispersity throughout eukaryotic genomes. Mass spectrometry is rapidly becoming a significant method in DNA analysis and has high probability of revolutionizing the way in which scientists probe the human genome. It is our responsibility as biomolecular mass spectrometrists to understand the issues in genetic analysis and the capabilities of mass spectrometry so that we may fulfill our role in developing a rapid, reliable technology to answer specific biological questions. This perspective is intended to familiarize the mass spectrometry community with modern genomics and to report on the current state of mass spectrometry, specifically electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry, for characterization of STRs. PMID- 11433533 TI - Determination of the electron affinities of alpha- and beta-naphthyl radicals using the kinetic method with full entropy analysis. The C-H bond dissociation energies of naphthalene. AB - The C - H bond dissociation energies for naphthalene were determined using a negative ion thermochemical cycle involving the gas-phase acidity (Delta H (acid)) and electron affinity (EA) for both the alpha- and beta-positions. The gas-phase acidity of the naphthalene alpha- and beta-positions and the EAs of the alpha- and beta-naphthyl radicals were measured in the gas phase in a flowing after glow-triple quadrupole apparatus. A variation of the Cooks kinetic method was used to measure the EAs of the naphthyl radicals by collision-induced dissociation of the corresponding alpha- and beta-naphthylsulfinate adducts formed by reactions in the flow tube portion of the instrument. Calibration references included both pi and sigma radicals, and full entropy analysis was performed over a series of calibration curves measured at collision energies ranging from 3.5 to 8 eV (center-of-mass). The measured EAs are 33.0 +/- 1.4 and 31.4 +/- 1.0 kcal mol(-1) (1 kcal = 4.184 kJ) for the alpha- and beta-naphthyl radicals, respectively. The gas-phase acidities for naphthalene were measured by the DePuy silane cleavage method, which utilizes the relative abundances of aryldimethylsiloxides and trimethylsiloxide that result from competitive cleavages from a proposed penta coordinate hydroxysiliconate intermediate. The measured acidities are 394.0 +/- 5.0 and 397.6 +/- 4.8 kcal mol(-1) for the alpha and beta- positions, respectively. The C - H bond dissociation energies calculated from the thermochemical cycle are 113.4 +/- 5.2 and 115.4 +/- 4.9 kcal mol(-1) for the alpha- and beta-positions, respectively. These energies are, to within experimental error, indistinguishable and are approximately the same as the first bond dissociation energy for benzene. PMID- 11433534 TI - Investigation of tyrosine nitration in proteins by mass spectrometry. AB - In vivo nitration of tyrosine residues is a post-translational modification mediated by peroxynitrite that may be involved in a number of diseases. The aim of this study was to evaluate possibilities for site-specific detection of tyrosine nitration by mass spectrometry. Angiotensin II and bovine serum albumin (BSA) nitrated with tetranitromethane (TNM) were used as model compounds. Three strategies were investigated: (i) analysis of single peptides and protein digests by matrix-assisted laser desorption/ionization (MALDI) peptide mass mapping, (ii) peptide mass mapping by electrospray ionization (ESI) mass spectrometry and (iii) screening for nitration by selective detection of the immonium ion of nitrotyrosine by precursor ion scanning with subsequent sequencing of the modified peptides. The MALDI time-of-flight mass spectrum of nitrated angiotensin II showed an unexpected prompt fragmentation involving the nitro group, in contrast to ESI-MS, where no fragmentation of nitrated angiotensin II was observed. The ESI mass spectra showed that mono- and dinitrated angiotensin II were obtained after treatment with TNM. ESI-MS/MS revealed that the mononitrated angiotensin II was nitrated on the side-chain of tyrosine. The dinitrated angiotensin II contained two nitro groups on the tyrosine residue. Nitration of BSA was confirmed by Western blotting with an antibody against nitrotyrosine and the sites for nitration were investigated by peptide mass mapping after in-gel digestion. Direct mass mapping by ESI revealed that two peptides were nitrated. Precursor ion scanning for the immonium ion for nitrotyrosine revealed two additional partially nitrated peptides. Based on the studies with the two model compounds, we suggest that the investigation of in vivo nitration of tyrosine and identification of nitrated peptides might be performed by precursor ion scanning for the specific immonium ion at m/z 181.06 combined with ESI-MS/MS for identification of the specific nitration sites. PMID- 11433535 TI - Matrix-assisted laser desorption/ionization, fast atom bombardment and plasma desorption mass spectrometry of polyethylene glycol esters of (2-benzothiazolon-3 yl)acetic acid. AB - Fast atom bombardment (FAB), matrix-assisted laser desorption/ionization time-of flight (MALDI-TOF) and plasma desorption (PD) mass spectra of newly synthesized polyethylene glycols (PEGs), (M(w) 600-4000 Da) chemically modified with biologically active (2-benzothiazolon-3-yl)acetyl end-groups are described (products 1-6). The spectra were also used for the determination of the molecular mass characteristics (number average (M(n)) and weight average (M(w)) molecular masses) of the initial and modified PEGs. As expected, M(n) and M(w) of the modified samples are higher than those of the non-modified samples. However, it is shown that molecular mass dispersity (determined by the comparison of the polydispersity indices (PDI = M(w)/M(n)) of both types of PEGs) essentially do not change during this modification. The FAB mass spectra, together with molecular species, show the presence of abundant [M + Na](+) ions of product 1 and [M + Na + H](+) species of 2 and 3, and [M + Na + 2H](+) of product 4. Two main series of fragment ions, derived from the cleavage of the ether bonds, are observed. The number fractions of the molecular adduct ions and fragment adduct ions, determined from the FAB and PD mass spectra of the modified PEGs, are compared. The MALDI-TOF mass spectra of compounds 1-6 show the presence of two series of polymers. The most abundant peaks are due to [M + Na](+) and [M + K](+) ions originating from the polymers, in which the two terminal hydroxyl groups of PEGs are esterified with (2-benzothiazolon-3-yl)acetic acid. The less abundant peaks are due to the monosubstituted polymers. PMID- 11433536 TI - Spontaneous N --> O acyl shift in the [M + H](+) ions of [MeBmt1]-cyclosporins in an ion trap. AB - In an ion trap the protonated molecules of the cyclic undecapeptides cyclosporins having 3-hydroxy-4-methyl-2-methylamino-6-octenoic acid (MeBmt) in their backbone undergo an N --> O peptidyl shift into the corresponding [M + H](+) ions of isocyclosporins. This rearrangement does not take place in cyclosporins [Bmt1]Cs and [3'-deoxy-MeBmt1]Cs. In cyclosporin [Thr2]Cs having two threonines in the molecule, only one of them participates in the N,O-acyl transfer. It can be concluded that the presence of the basic N-methylamino group of MeBmt, which can serve as the primary site of protonation, is necessary for isocyclosporin formation. A dominating ion series originating from the primary cleavage between MeBmt (first position in the cyclosporin ring) and amino acid residue at the neighbouring eleventh position is then observed in collision-induced dissociation spectra of protonated molecules of cyclosporins. This 'isocyclosporin' ion series can effectively be used for easy and complete cyclosporin sequencing using a tandem mass spectrometric (MS3) experiment in an ion trap. The paper further introduces an improved Gross mass spectral nomenclature for cyclic peptide sequencing and several techniques for the generation of protonated molecules of cyclosporins. Their preparation represents the fundamental requirement for smooth sequencing of cyclosporins by tandem mass spectral techniques. PMID- 11433537 TI - Capabilities of static TOF-SIMS in the differentiation of first-row transition metal oxides. AB - Time-of-flight secondary ion mass spectrometry (TOF-SIMS) analyses were performed on the first-row transition metal oxides from scandium to zinc in positive and negative detection modes. The nature of the numerous M(x)O(y)(+/-) ionic species generated by 15 keV Ga(+) primary ion bombardment allows the identification of a given metal-oxygen system. To identify the metal valence in the oxide under investigation, several procedures were investigated: the detection of specific and characteristic ions, the use of ion abundance ratios and the use of a valence model. Owing to their importance in many fields of materials science, each of these speciation methodologies was evaluated for the differentiation of vanadium, titanium, chromium, manganese, iron, cobalt and copper oxides. Trivalent hexavalent chromium distinction was first intensely investigated because it really corresponds to a model system for inorganic speciation. For each series of metal oxides, the more pertinent speciation criteria were then systematically tested. The limitations of the proposed methodologies are discussed. Their use is made complicated when pollutants or a superficial oxide layer, with a stoichiometry different from that of the bulk, are present. Finally, thermodynamic considerations relative to the stability of the M(x)O(y)(+/-) ions may also modify the relationship between the analyzed oxide and the observed positive and negative secondary ion mass spectra. PMID- 11433538 TI - Determination and characterization of diuretics in human urine by liquid chromatography coupled to pneumatically assisted electrospray ionization mass spectrometry. AB - The aim of this work was to develop a method for the characterization and determination of diuretics in human urine samples by liquid chromatography (LC) coupled to pneumatically assisted electrospray ionization (ES) mass spectrometry (MS). The diuretics studied were substances forbidden by the IOC such as trichlormethiazide, furosemide, canrenoic acid, benzthiazide, bendroflumethiazide, bumetanide, etacrynic acid and spironolactone. For this purpose, the operational parameters of electrospray, such as counter electrode voltage, capillary voltage, sample cone voltage and source temperature, were optimized in order to obtain the best signal stability and the highest sensitivity for the greatest number of diuretic agents. The optimized separation method was successfully coupled with the MS system to analyze the above-mentioned diuretics extracted from spiked urine samples by a liquid extraction and clean-up procedure at basic pH, using ethyl acetate as solvent and the salting-out effect (NaCl). The mass spectra obtained provide adequate information for identification purposes. Positive urine samples obtained from athletes were also analyzed. The presence of these substances in human urine was confirmed by this method, making LC/ES-MS an analytical tool to be considered in the area of antidoping control. PMID- 11433539 TI - Sequencing cyclic peptide inhibitors of mammalian ribonucleotide reductase by electrospray ionization mass spectrometry. AB - Mammalian ribonucleotide reductase (mRR) is a potential target for cancer intervention. A series of lactam-bridged cyclic peptide inhibitors (1-9) of mRR have been synthesized and tested in previous work. These inhibitors consist of cyclic and linear regions, causing their mass spectral characterization to be a challenge. We determined the fragmentation mechanism of cyclic peptides 1-9 using an ion-trap mass spectrometer equipped with an ESI source. Low-energy collision induced dissociation of sodiated cyclic peptides containing linear branches follows a general pathway. Fragmentation of the linear peptide region produced mainly a and b ions. The ring peptide region was more stable and ring opening required higher collision energy, mainly occurring at the amide bond adjacent to the lactam bridge. The sodium ion, which bound to the carbonyl oxygen of the lactam bridge, acted as a fixed charge site and directed a charge-remote, sequence-specific fragmentation of the ring-opened peptide. Amino acid residues were cleaved sequentially from the C-terminus to the N-terminus. Our findings have established a new way to sequence cyclic peptides containing a lactam bridge based on charge-remote fragmentation. This methodology will permit unambiguous identification of high-affinity ligands within cyclic peptide libraries. PMID- 11433540 TI - On-line coupling of hollow fiber membranes with electrospray ionization mass spectrometry for continuous affinity selection, concentration and identification of small-molecule libraries. AB - Combinatorial chemistry has been widely employed in the pharmaceutical industry in the effort towards drug discovery. Rapid and sensitive screening of lead candidates among library compounds has thus imposed significant analytical challenges in recent years. This work involved the development of a continuous affinity capture and concentration system, providing cost-effective and structural analysis of drug candidates in a flow-through format. The system combines the strengths of a hollow fiber dialysis membrane of ease and speed of purification and concentration with the specificity of affinity interactions in solution. The complexes between the lead compounds and the affinity binding proteins are separated from other chemical components inside a dialysis hollow fiber as the result of their differences in size. The affinity complexes are further concentrated inside a second dialysis fiber. The concentrated drug candidates are liberated from the binding proteins in a microdialysis junction and can be directly identified using electrospray ionization mass spectrometry. Two model systems, including human serum albumin-warfarin-related compounds and anti-phenobarbital antibody-barbiturates, were employed for mechanistic studies of dialysis versus dissociation kinetics and competitive selection of drug candidates according to their binding strengths. PMID- 11433541 TI - Quantitation of cyclosporin A in whole blood by liquid chromatography/stable isotope dilution electrospray ionization mass spectrometry. AB - Therapy with cyclosporin A (CsA) for immunosuppression after organ transplantation requires monitoring of its levels in blood owing to the narrow therapeutic index of the drug and to the high inter-individual variability of the drug absorption and metabolism. We describe the preparation of CsA labelled with stable isotopes ((13)C and (2)H) with an isotopic enrichment of about 99% using labelled glucose and its use as internal standard for quantification of CsA blood levels by isotope dilution/electrospray ionization mass spectrometry. The method was found to be linear in the tested range (1-1000 ng) with and without the matrix. The accuracy of the bracketting calibration curves prepared using 100 ng ml(-1) labelled CsA was within +/-1.7% (bias). The results confirmed the usefulness of the procedure as a reference method for the external quality assessment of the field methods for the evaluation of CsA blood concentration, the imprecision (relative standard deviation) and accuracy (bias) being <2%. PMID- 11433542 TI - Electrospray ionization mass spectrometric studies on the amphoteric surfactant cocamidopropylbetaine. AB - After liquid chromatographic (LC) separation, electrospray ionization mass spectrometry (ESI-MS) was investigated for the determination of the amphoteric surfactant cocamidopropylbetaine (CAPB). In the positive ion mode the molecule formed the adduct ions [M + H](+), [M + Na](+) and [M + K](+). Adducts of these cations were also detected with decreasing abundance as dimer and trimer clusters. Additionally, doubly charged molecular ions with different combinations of cations were identified. It was noticed that the relative abundances of individual cation adducts were not reproducible, apparently owing to varying contents of alkali metal ions originating from the solvent and the sample. Under negative ionization, the major molecular ion was [M - H](-). Higher clusters formed by two and three surfactant molecules, i.e. [2M - H](-) and [3M - H](-) were likewise registered. The tendency to form clusters in both positive and negative ion modes, even at 0.1 mg l(-1) levels, was attributed to strong electrostatic interactions between the zwitterionic head groups. Further evidence for this assumption was provided by the detection of a fragment formed from [2M - H](-) which contained the two charged head groups. Studies were undertaken in the negative ion mode on the concentration- and orifice voltage-dependent monomer, dimer and trimer formation of C(12)-CAPB in order to evaluate potential issues in using the ion [M - H](-) mode for quantitative analysis. Finally, the established (-)-LC/ESI-MS method was applied to follow up the primary degradation of CAPB in a laboratory-scale fixed-bed bioreactor (FBBR) spiked with a test concentration of 10 mg l(-1). Direct analysis without sample pretreatment revealed that higher alkyl homologues were more prone to adsorption. Primary biodegradation of all alkyl homologues was completed after a period of 4 days. Selected lyophilized FBBR samples were examined for the presence of transient or stable degradation intermediates, but no metabolite could be identified. PMID- 11433543 TI - Formation of *CH2OC(O(-))CR2 distonic radical anions by the reaction of O(-.) with dimethyl glutarates. PMID- 11433545 TI - An intelligent tele-healthcare environment offering person-centric and wellness maintenance services. AB - Worldwide healthcare delivery trends are undergoing a subtle paradigm shift- patient centered services as opposed to provider centered services and wellness maintenance as opposed to illness management. In this paper we present a Tele Healthcare project TIDE--Tele-Healthcare Information and Diagnostic Environment. TIDE manifests an 'intelligent' healthcare environment that aims to ensure lifelong coverage of person-specific health maintenance decision-support services -i.e., both wellness maintenance and illness management services--ubiquitously available via the Internet/WWW. Taking on an all-encompassing health maintenance role--spanning from wellness to illness issues--the functionality of TIDE involves the generation and delivery of (a) Personalized, Pro-active, Persistent, Perpetual, and Present wellness maintenance services, and (b) remote diagnostic services for managing noncritical illnesses. Technically, TIDE is an amalgamation of diverse computer technologies--Artificial Intelligence, Internet, Multimedia, Databases, and Medical Informatics--to implement a sophisticated healthcare delivery infostructure. PMID- 11433546 TI - An approach for integrating heterogeneous information sources in a medical data warehouse. AB - In recent years, medical professionals are witnessing an explosive growth in data collected by various organizations and institutions. At the same time, the ongoing developments of networking technologies provide doctor with the capability to access these data across the boundaries of interconnected computers. In this paper we present a medical data warehousing methodology that aims to use data semantics to regroup and merge patients' medical data from different health information systems, which may be autonomous and heterogeneous. The proposed solution takes into account European laws concerning the security and anonymity of personal data. PMID- 11433547 TI - Tools for acquisition, processing and knowledge-based diagnostic of the electroencephalogram and visual evoked potentials. AB - The objective of our research is to develop computer-based tools to automate the clinical evaluation of the electroencephalogram (EEG) and visual evoked potentials (VEP). This paper describes a set of solutions to support all the aspects regarding the standard procedures of the diagnosis in neurophysiology, including: (1) acquisition and real-time processing and compression of EEG and VEP signals, (2) real-time brain mapping of spectral powers, (3) classifier design, (4) automatic detection of morphologies through supervised neural networks. (5) signal analysis through fuzzy modelling, and (6) a knowledge based approach to classifier design. PMID- 11433548 TI - Towards more optimal medical diagnosing with evolutionary algorithms. AB - Efficiency in hospital performance is becoming more and more important. Studies showed that diagnosis can considerably reduce the inefficiency, so one of the most important tasks in achieving greater hospital efficiency is to optimize the diagnostic process. For the best of the patient the diagnostic process has to be optimized regarding the number of the examinations and individualized in order to maximize accuracy, sensitivity and specificity. In addition the duration of the diagnostic process has to be minimized and the process has to be performed on the most reliable equipment. The main contribution of our paper is the introduction of the integrated computerized environment DIAPRO enabling the diagnostic process optimization. The DIAPRO is based on a single approach--evolutionary algorithms. PMID- 11433549 TI - Artificial life simulation of living alga cells and its sorption mechanisms. AB - Resistance mechanisms of organisms against toxic metals are based on a few different mechanisms provided by algae cells. These mechanisms can be localized on the cell wall, on the cell wall and cytoplasm membrane, and intracellular localized mechanisms. Due to these mechanisms, algae populations could be used for sorption of arsenic from contaminated waters. This process takes a long time and it's not very variable. With artificial life simulation based on multi-agent simulation system we are preparing a simulation that could at least partially substitute the real experiments with real cells. Artificial life simulation of alga Chlorella kessleri is based on real biological parameters and together with partial implementation of other mathematical models of algae population growth it will be used for simulating the process of absorbing heavy metals from contaminated water. Model implementation is done is Swarm--multi agent object based simulation system and it's libraries. Simulation is still in testing and debugging phase and it is not yet available for public. PMID- 11433550 TI - Implementation of Web-based interaction technology to improve the quality of a city's health care. AB - In the clinic, one method for improving the interaction is to ask patients to systematically report their health status, give them standard advice based on their responses, and ask them to discuss this advice with a health practitioner. In the school system, this approach provides aggregate information for targeting programs to meet student needs. In the workplace, this health assessment and personal feedback approach may be offered to employees to improve health care and lower health care costs. But why stop at the door of the clinic, school, or workplace when Internet technology can extend to an entire community the benefits of health assessment and feedback? PMID- 11433551 TI - Stanford Health Partners: rationale and early experiences in establishing physician group visits and chronic disease self-management workshops. AB - Chronic disease is a major public health problem. The day-to-day management of a chronic illness requires accurate patient reporting and timely medical response. Patient group visits with the primary care physician and lay-led patient education workshops improve the provider-physician relationship, promote patient self-efficacy and self-management of chronic diseases, and facilitate positive health care outcomes. This article describes how group visits are being integrated into the clinical routine of a health care system. PMID- 11433552 TI - How the health care system can improve mammography-screening rates for underserved women: a closer look at the health care delivery system. AB - The way care is delivered has dramatic impact on the patient-provider interaction and the outcomes experienced by the patient. This article explores a deceptively simple but very powerful method for evaluating and improving care delivery. Mammography is a routine screening procedure. However, many factors can influence how frequently women seek and obtain mammograms. Twenty-five low-income women identified empowering factors and barriers they experienced when trying to obtain a mammogram. PMID- 11433553 TI - Merging education with measurement: a focus on the Hispanic community. AB - There is an increasing need for community health promotion efforts directed to populations that are often disenfranchised from the health care system. The rapidly growing Hispanic population can benefit from outreach efforts that provide health education and promote community awareness of health status and health problems. This article describes a cost effective survey system that measured health status, health needs and provided individualized health feedback and education to Hispanic residents. The results underscore the importance of culturally and linguistically appropriate community-based initiatives that assess health status to aid in setting policy, eliminate health disparities, and determine priorities to meet the health needs of the growing Hispanic population. PMID- 11433554 TI - African American perspectives on health care: the voice of the community. AB - African Americans often have interactions with health care systems, services, and providers that are quite different from those of other population groups. Residents in a predominantly African American community in the southeastern United States identified issues that had a significant influence on their health care interactions. Their insights about health insurance concerns, perceived quality of health care, and trust in the health care system provide a useful framework for the redesign of care that will better meet their health care needs. PMID- 11433555 TI - Correctional health care as a vital part of community health. AB - Correctional facilities afford health professionals an opportunity to serve a segment of society that may not otherwise come into contact with the health care system. This article illustrates why it is imperative that screenings for infectious diseases be conducted in correctional facilities and/or treatment administered before detainees are released back into the general population. Correctional facilities should be included as alternative health stations to improve community health. PMID- 11433556 TI - Use of innovative technologies in the evaluation of Nashville's REACH 2010 community action plan: reducing disparities in cardiovascular disease and diabetes in the African American community. AB - Ethnic minorities in the United States suffer disproportionately from chronic diseases such as cancer, heart disease, AIDS, and diabetes. A nationally-funded initiative known as Racial and Ethnic Approaches to Community Health (REACH) has designated 26 communities to implement demonstration projects to reduce health disparities in targeted minority populations. This article presents two methods of integrating innovative technologies into the program's evaluation plan: (1) a Web-based data entry system for recording project activity and (2) geographic information systems (GISs) for developing visual maps of project targets. These technologies can assist other programs in managing and measuring community-based public health initiatives. PMID- 11433557 TI - Addressing challenges, creating opportunities: fostering consumer participation in Medicaid and Children's Health Insurance managed care Programs. AB - With more than half of the Medicaid population enrolled in managed care and with enrollment in Medicaid managed care and Children's Health Insurance Programs continuing to accelerate, policy makers and program administrators must quickly come to consensus on the role of the consumer in the planning, implementation, and oversight of Medicaid and S-CHIP managed care. This article explores the barriers to consumer participation and makes concrete policy and programmatic recommendations to increase consumer participation in public managed care programs. To fully comply with existing federal and state requirements for consumer participation and for this participation to have an impact on the quality of managed care programs, states need to provide more education and support to consumers on: how to choose a health plan; how to access the health plan services and out-of-network benefits; and how to use grievance procedures. In addition, more work needs to be done to make the growing availability of health plan and provider performance data meaningful and available to Medicaid and S-CHIP consumers. Finally, states must address the lack of resources, lack of information, and lack of training that prevent many consumers from being able to sustain their involvement in system-level advisory and oversight bodies. PMID- 11433558 TI - Health status assessments using the Veterans SF-12 and SF-36: methods for evaluating otucomes in the Veterans Health Administration. AB - This article describes a study in which the administration of two health surveys, the Veterans SF-36 and SF-12, by telephone and mail-out was used to assess the differences in the health surveys' costs and scores by mode of administration and determine which mode was cost-efficient. The study employed a crossover design: after 12 unsuccessful attempts to contact patients by telephone, the patients were administered the survey by mail, and after 2 unsuccessful mail-outs, up to three attempts were made to interview the patients by telephone. The analysis of the data showed that mail administration, with or without crossover to telephone, was more cost-efficient than telephone administration, having both lower average total and variable costs per completed questionnaire. Overall, telephone administration was about 30% more expensive that mail administration, primarily due to the cost of labor. The marginal cost of an additional completed Veterans SF-12 or Veterans SF-36 was also substantially lower for mail administration. Mail administration without crossover to telephone administration was the most cost-efficient strategy for administering both the Veterans SF-12 and SF-36. The results of this study strongly suggest the need to consider the mode of administration if questionnaires like the Veterans SF-12 or SF-36 are to be used to assess health outcomes within and across large health care systems. PMID- 11433559 TI - [Is the spiral CT today the benchmark for diagnosing pulmonary embolism?]. PMID- 11433560 TI - [Cat scratch disease. Clinical study of 26 patients. Role and importance of PCR]. AB - PURPOSE: Cat scratch disease is a mild pathology but diagnosis often remain difficult. METHODS: A retrospective study has been conducted by the department of infectious diseases at the University hospital in Angers. Between January 1994 and October 1998, 26 observations were recorded providing the presence of three criteria out of four among the following: contact with a cat, clinical presentation and its favorable course, absence of any other cause noticed, and the presence of either a positive serology or a positive PCR, or the examination of a suggestive pathology. RESULTS: Fourteen men and 12 women were concerned. From a clinical point of view, the inoculation lesion was observed six times, all patients showed at least one adenopathy during their illness, 12 patients showed only an adenopathy without clinical signs. A surgical biopsy was carried out on nine patients and a diagnosis established. Bartonella henselae serology was done in all patients. Six presented a significant rate of IgG antibodies as early as the first dosage. A seroconversion was observed in four cases belatedly 1 to 2 months after the beginning of the symptomatology. The method's sensitivity was approximately 38%. A PCR search was accomplished in the pus obtained from a ganglionic puncture on 12 patients. It was positive seven times, which corresponds to a sensitivity of about 58%. In associating these two diagnostic criteria a sensitivity rate of nearly 92% was reached, the diagnosis not having been confirmed only in one case. The outcome proved to be favorable in all cases, with or without an antibiotic treatment. CONCLUSION: The association of serology and PCR in the pus permits a certain diagnosis in the majority of the cases and avoids the more aggressive biopsy. PMID- 11433561 TI - [An index based on the age and free thyroxine rate predicts the likelihood of Graves' disease in hyperthyroidism]. AB - PURPOSE: Graves' disease patients are generally younger and have more severe symptoms than other thyrotoxic patients. We established an index based on the normalized free thyroxine rate and age, capable of predicting Graves' disease in thyrotoxic patients. METHODS: The predictive index was established from a discriminant analysis from a retrospective population of 114 thyrotoxic patients and its predictivity was confirmed by cross-validation on the same population. RESULTS: The index IGD = 41.38 - age + 37.05 x in (normalized free T4) classifies accurately 80.7% of the patients (CI 95%: 72.2-87.5). Sensitivity is 78.5% (CI 95%: 66.5-87.7). Specificity is 83.7% (CI 95%: 70.3-92.7). Predictive positive value is 86.4% (CI 95%: 75.0-94.0). Negative predictive value is 74.5% (CI 95%: 61.0-85.3). Likelihood ratio of Graves' disease in case of prediction by the index is 4.81 (CI 95%: 2.66-9.32). Likelihood ratio of Graves' disease in case of non-prediction by the index is 0.26 (IC 95%: 0.16-0.40). CONCLUSION: A simple index based on the normalized free thyroxine rate and age provides an early diagnosis orientation toward Graves' disease in thyrotoxic patients who are waiting for complementary investigations. PMID- 11433562 TI - [Acute colonic pseudo-obstruction in internal medicine: etiology and prognosis, report of a retrospective study]. AB - PURPOSE: To analyze the context in which acute colonic pseudo-obstructions occur in a medical practice. METHOD: Retrospective study of patients with an acute colonic pseudo-obstruction hospitalized in an internal medicine department between November 1992 and December 1999. RESULTS: We found 12 patients including eight men (mean age: 80.2 years). All had abdominal pains. Seven patients presented an occlusive syndrome. A septic shock occurred in two subjects. The average diameter of the cecum was 10 cm. Distention involved the whole colon for nine patients, and only the left part for three. Six subjects were affected by a bronchopneumonia, three had a previous history of recent diarrhea, two had colonic diverticulitis, and two others had a diabetic neuropathy. Ogilvie's syndrome revealed hypothyroidism for one patient. Nine patients were taking one or more medications incriminated in the acute colonic pseudo-obstruction: calcium channel blockers (n = 6), neuroleptics (n = 5), antidepressant agents (n = 4), dopaminergic drugs (n = 2) and opiates (n = 1). Five patients underwent a colonoscopic decompression. Surgical procedures were undertaken for three patients: one underwent a complete colectomy because of numerous perforation. Except for the colectomized patient, the outcome resulted in either recovery (n = 7), partial improvement (n = 3), or recurrence (n = 1). CONCLUSION: In a medical environment, acute colonic pseudo-obstruction is a pathology for elderly persons, namely with a masculine predominance and often associated with iatrogenic factors. The outcome is more often than not favorable, but it may result in invasive acts or significant complications at this age. PMID- 11433563 TI - [Tick-born encephalitis]. AB - INTRODUCTION: Tick-borne encephalitis (TBE), a disease contracted through tick bites, is caused by a Flavivirus. Its geographical distribution comes from the geographical distribution of the reservoir of infection--i.e., mainly the tiny mammals living in the forests and bushes. The endemic area spreads from the Rhine to the Urals, from Scandinavia to Italy and Greece. CURRENT KNOWLEDGE AND KEY POINTS: Symptoms usually evolve in three phases: at first a nonspecific phase with fever and myalgia, then an afebrile phase, and finally a phase with neurological manifestations, such as meningitis, meningoencephalitis and/or myelitis, and fever. Motor neurological sequelae are possible. The cases occurring in the East are characterized by their greater severity compared to those occurring in the West. The diagnosis, easily established given a history of a tick bite in an endemic area, is confirmed by the presence of specific IgM in the blood and/or cerebral spinal fluid. FUTURE PROSPECT AND PROJECTS: There is no specific treatment. Prevention consists of individual prophylactic measures (self examination and systematic extraction of ticks after exposure, use of repellents), and in immunization. The vaccine, prepared from inactivated viruses, should be used for target populations, that is, for people exposed to tick bites during their professional or leisure outdoor activities. PMID- 11433564 TI - [Current methods for diagnosing renal artery stenosis]. AB - PURPOSE: Today, the methods for detecting renal artery stenosis are numerous. They lead to the identification either of anatomic stenosis or functional stenosis (onset of renin-angiotensin system). The purpose of this review is to emphasize the performances, limitations and diagnostic criteria for each technique. CURRENT KNOWLEDGE AND KEY POINTS: For detection of anatomic stenosis, the feasibility of Doppler examination has been increased by the improvement of US systems and the development of contrast agents. However, the criteria used nowadays still need wider evaluation. Helical CT angiography allows a better detection of accessory arteries and calcifications but MR angiography has a better spatial resolution and uses a non-nephrotoxic contrast agent. The performances of these two techniques have never been compared. The choice between these techniques depends on the operators' experience and the availability of the systems. For functional stenosis, detection is now based on captopril-sensitized scintigraphy. FUTURE PROSPECTS AND PROJECTS: It would be necessary to better compare performances and cost-effectiveness ratios of these methods and of the two imaging strategies (detection of anatomic stenosis versus functional stenoses). Probably in the future, MR will gain a major place, providing, in the same imaging session, morphologic and functional data. PMID- 11433565 TI - [Disclosure of a diagnosis of cancer: paradoxes and misunderstandings]. AB - INTRODUCTION: Announcing to a patient that he has cancer is a key moment both in the relationship between this patient and his doctor and in the setting up of quality communications. However, the quality of this communication is often impaired by the presence of paradoxes and misunderstandings. CURRENT KNOWLEDGE AND KEY POINTS: The particularities of cancer, the defense mechanisms of the different contributors and cancer representations can increase the sources of misunderstandings, which will leave an indelible mark on the patient at the beginning of the disease. FUTURE AND PROJECTS: What's at stake in that type of announcement in terms of compliance should not be neglected. Practitioners becoming aware of those phenomena and communication rules should improve the modes of announcement. PMID- 11433566 TI - [Vertebral actinomycosis with spinal cord compression. A case report]. AB - INTRODUCTION: Vertebral involvement of actinomycosis is extremely rare and associated spinal cord compression is unusual. EXEGESIS: We report a case of a 31 year-old man with vertebral actinomycosis presenting with spinal cord compression. Magnetic resonance imaging demonstrated a paravertebral abscess and lytic areas on the vertebral body of C5 requiring emergency surgery and antibiotic treatment. The patient was still asymptomatic after 12 months of follow-up. CONCLUSION: Clinical aspects of this unusual localization are reviewed. Treatment may be particularly difficult. PMID- 11433567 TI - [Natural killer cell nasal lymphoma mimicking localized Wegener's disease]. AB - INTRODUCTION: Primary non-Hodgkin's lymphoma of the nasal cavity is particular. Pathological characteristics mainly associate a prevalent NK lymphocyte phenotype, a frequent exposure to the Epstein-Barr virus and a poor sensitivity to radiotherapy compared to other lymph node localizations. EXEGESIS: The authors report the case of a 38-year-old man. The patient had previously presented a chronic maxillary sinusitis. After a diagnosis of Wegener's disease, the poor course under therapy resulted in a nasal lymphoma. Natural killer cell nasal lymphoma was confirmed with a leading biopsy at the same time as a serious clinical outcome. The patient died of septic shock with multivisceral failure. CONCLUSION: The two differential diagnoses of ulcerative lymphoma of the midface are ulcerative infectious diseases and Wegener's disease. We must not miss this severe disease, with its poor prognosis and variable, though sometimes rapid speed of evolution. PMID- 11433568 TI - [Pubic osteomyelitis in athletes]. AB - INTRODUCTION: Pubic osteomyelitis has been described in three situations: children in whom Staphylococcus aureus is the preeminent pathogen; elderly patients who have undergone genitourinary procedures, and parenteral drug abusers. In contrast, pubic osteomyelitis in athletes has been described less often. We report three cases of acute staphylococcal pubic osteomyelitis in young athletic men and present a review of the literature. EXEGESIS: The clinical presentation in each case was acute groin, hip, or perineal pain; fever; inability to bear weight; and pubic symphysis tenderness. The diagnosis was established by blood culture and radiologic changes. CONCLUSIONS: Staphylococcus aureus pubic osteomyelitis should be suspected in athletes who have febrile hip or groin pain. The pathogenesis of this disease is thought to involve preexisting trauma or athletic injury and subsequent seeding of this area during transient bacteremia. Prolonged antimicrobial therapy is required for the cure, and debridement with curettage may be necessary if patients have persistent infection or sequestra. PMID- 11433569 TI - [DRESS (Drug Rash with Eosinophilia and Systemic Symptoms) syndrome after sulfasalazine and carmazepine: report of two cases]. AB - INTRODUCTION: To better individualize drug hypersensitivity reaction, Bocquet et al. have recently called this adverse drug reaction DRESS (Drug Rash with Eosinophilia and Systemic Symptoms). EXEGESIS: We report two cases of DRESS and highlight the main characteristics of this syndrome. Two patients presented severe febrile skin eruption following drug intake (carbamazepine or sulfazalazine), with hypereosinophilia and hepatitis. All symptoms resolved after drug withdrawal and corticosteroid therapy. DRESS syndrome is an idiosyncratic reaction characterised by febrile eruption, occurring 2 to 6 weeks after the beginning of the treatment, accompanied by systemic symptoms and biological abnormalities (hypereosinophilia, hepatitis). Some complications can occur. This syndrome can be fatal. Numerous drugs can be responsible for this reaction to medication. The physiopathology has not yet been elucidated, and the treatment is not codified, but the triggering agent must immediately be stopped. Corticotherapy is sometimes used. CONCLUSION: It is important to recognize this entity recently named DRESS syndrome because it can mimic other pathologies, is potentially serious, and because withdrawal of the incriminating drug is imperative. PMID- 11433570 TI - [A troublesome foot]. PMID- 11433571 TI - [Encephalopathy during cat scratch disease in an adult]. PMID- 11433572 TI - ["Catastrophic antiphospholipid syndrome: a new case with favorable outcome]. PMID- 11433573 TI - [Chronic polyradiculoneuropathy in a patient with Hodgkin's disease]. PMID- 11433574 TI - ["Wandering" spleen revealed by hematemesis]. PMID- 11433575 TI - [A strange disease of the plateaux. Visceral leishmaniasis]. PMID- 11433576 TI - [A dog's life!...latent syphilis]. PMID- 11433577 TI - [Aphthous ulcers have their secrets....]. PMID- 11433578 TI - [An exhausted small intestine....Susac syndrome]. PMID- 11433579 TI - [Eggs in a fritter....progressive multifocal leukoencephalopathy]. PMID- 11433580 TI - [What hell!....pulmonary alveoli hemorrhage]. PMID- 11433581 TI - [A heart in springtime. Congestive heart failure and Osler Rendu disease]. PMID- 11433582 TI - [It's long, like lacunas! Actinomycosis bone infection]. PMID- 11433583 TI - [A dear old lady not so crazy as this....bismuth poisoning]. PMID- 11433584 TI - [Myoclonic coma and severe cardiac insufficiency in a patient with arteritis]. PMID- 11433585 TI - [An Antilles fever. Dapsone allergy]. PMID- 11433586 TI - [The sixth case....parathyroid adenoma]. PMID- 11433587 TI - [Stickler's syndrome (dystrophia vitreoretinalis hereditaria). Results of surgery for retinal detachment]. AB - OBJECTIVE: Stickler's syndrome (SS) is an autosomal dominant hereditary disease of the collagenous connective tissue where impaired development of the vitreous body gel and peripheral retina and detachment of the retina are associated with general manifestations. The objective of the retrospective study was to evaluate the long-term results of surgery of retinal detachment in SS. PATIENTS: The group of patients comprised 7 patients, 6 men and 1 woman aged 4 to 45 years, average age 16.8 years. Autosomal dominant heredity was obvious in 6 members (85.7%) of two families. General manifestations of SS included abnormalities of the facial skeleton (6 patients), cleft palate (4 patients), impaired hearing (2 patients), marfanoid habitus (2 patients) and hyperextensibility of the joints (4 patients). In the eyes with SS was manifested by myopia from -1 to -9 D and a liquid vitreous body. Multiple foci of lattice degeneration supplemented the finding in 6 patients (85.7%). Detachment of the retina was a manifestation of SS in 12 of 14 eyes (85.7%). It was manifested in 5 of 7 patients concurrently or within 12 years in both eyes. The causes of retinal detachment were multiple equatorial and postequatorial tears due to lattice degeneration in 8 eyes (66.7%) or a giant tear in 4 eyes (33.3%). Advanced proliferative vitreoretinopathy (PVR) was associated with retinal detachment in 8 eyes (66.7%) and in 6 eyes (50%) it was not possible to assess the beginning of retinal detachment. RESULTS: In 3 of 5 patients with bilateral retinal detachment the adverse course of retinal detachment on the first eye was followed 8 to 12 years previously in another department: two retinal detachments with giant tears were evaluated as inoperable and one inveterated detachment with advanced PVR was operated unsuccessfully. Retinal detachment was operated in 9 eyes of 7 patients, in two patients both eyes were operated simultaneously. The patients were followed up after surgery for 11 months to 15 years, on average for 65 months. A cryosurgical approach was used in 6 retinal detachments (66.7%), accentuated cerclage with drainage of subretinal fluid in 5 and radial plombage in one eye. The retina attached in 5 of 6 eyes (83.3%) and the final visual acuity was 0.25-0.1, mean 0.66. Pars plana vitrectomy (PPV) and implantation of silicone oil (ISO) were the final surgical solution in 3 eyes of 3 patients (33.3%). In one eye with retinal detachment with a circumferential width of 320 degrees and advanced PVR PPV and ISO were the primary operation, in two eyes re-operation on account of a relapse of retinal detachment with advanced PVR after an unsuccessful cryosurgical operation. Anatomical and functional success was achieved in 2 of 3 eyes (66.7%), in one eye after removal of silicone oil from the eye. The final visual acuity of successfully operated eyes was 0.2 and 0.16. CONCLUSIONS: Retinal detachment in SS usually affects both eyes, the second eye may be affected after many years. The cause of retinal detachment are multiple tears or a giant tear. Early surgery of retinal detachment has as a rule a favourable effect, while late diagnosis of retinal detachment in children leads frequently to blindness. Dispensarization and regular ophthalmological check-ups of patients with SS are an essential prerequisite of successful prevention and treatment of retinal detachment. PMID- 11433588 TI - [Pars plana vitrectomy in treatment of tractional edema of the macula]. AB - We analysed the causes and the consequences of traction macular edema with the possibility of solution of this stage by pars plana vitrectomy with removal of vitreomacular traction and epiretinal macular membranes. An analysis of preoperative and postoperative findings of 4 patients was made. There were found idiopathic epiretinal membranes in 3 cases and vitreomacular traction syndrome in 1 case. We reported postoperative improvement of visual acuity in 3 cases, stabilisation of postoperative state was reached in 1 patient. PMID- 11433589 TI - [Initial experience with toric intraocular lenses]. AB - The aim of cataract extraction with implantation of intraocular lens is the astigmatically neutral result. This neutral cataract incision is reached by the method of phacoemulsification with a small incision and implantation of foldable intraocular lens. In case of preoperative astigmatism is an effective method the implantation of a toric intraocular lens. The authors evaluate the first results of pre and postoperative refraction after implantation of ten toric intraocular lenses. PMID- 11433590 TI - [Results of 32 cataract operations using the capsular ring]. AB - The authors present results of 32 operations with capsular tension ring performed in eyes with zonular dehiscence. There were 14 women (50%) and 14 men (50%), altogether 28 patients (32 eyes). Mean patient age was 47.1 years (range 22-89 years). The reason for zonular weakness was trauma in 15 eyes (46.9%), Marfan's syndrome in 8 eyes (25.0%), pseudoexfoliation syndrome in 6 eyes (18.7%) and high myopia in 3 eyes (9.4%). Anterior vitrectomy was done if the vitreous appeared in the anterior chamber. Early postoperative complications were fibrinoid reaction in 2 eyes (6.3%), intraocular pressure elevation in 5 eyes (15.6%), hyphaema in 2 eyes (6.3%). Best corrected visual acuity was 0.5 or better in 18 eyes (56.3%) on the first postoperative day and in 17 eyes (53.1%) at the time of the last examination. Our results have shown the high efficiency of the capsular tension ring in managing zonular problems during cataract surgery. PMID- 11433591 TI - [Duane's retraction syndrome--overview and diagnosis of clinical types]. AB - The author presents a postgraduate review of the problem. In the introduction he reviews typical features of Duane's retraction syndrome (DS) and its main symptoms and reminds of the main approaches to classification: (1) Malbrane s (Duane I, II and III), (2) Huber's which is based on EMG findings in ZOS and (3) Kaufman's which classifies DS according to the enforced position of the head. The author maintains that even according to the above many atypical rare pictures cannot be classified or explained pathogenetically. Some are mentioned: (1) "Inverse" DS, which was recorded and documented by Chytilova-Divisova (1949) in a girl with congenital paralysis of abduction on both eyes retraction of the bulbus developed and narrowing of the palpebral aperture when attempting abduction (1) of the eye, (2) Bilateral acquired DS in a female patient with a tumour of the brain stem confirmed by EMG records of both horizontal muscles. (3) Unilateral DS in a child from a family with familial incidence of congenital ZOS fibrosis with an obscure ratio of the neurogenic and myogenic and fibrous component of the two pictures. In another member of this family the Marcus Gunn phenomenon was present. The latter findings support the idea that in the development of the fairly uniform picture of DS a combination of neurogenic, myogenic and connective tissue changes participate. PMID- 11433592 TI - [Duane's retraction syndrome--surgical treatment]. AB - The author presents possible surgical approaches in Duane's syndrome I. The weakening operation of the medial rectus muscle by means of marginal myotomy is recommended in case of striking ocular torticolis. The bifurcation recession of the lateral rectus muscle is recommended in case of change of palpebral fissure. The author documents the indications and the after-surgery results. PMID- 11433593 TI - [Echography findings in orbital myositis]. AB - In a retrospective study comprising 12 patients with the diagnosis or orbital myositis the authors present an account on the importance of standardized echography for the differential diagnosis and for the follow-up of the disease in relation to treatment. They describe the sonographic finding in affected oculomotor muscles and in myositis of the upper oblique muscle incl. the echographic finding of trochleitis. The authors discuss echographic signs of myositis (thickness and shape of muscle, reflectivity and internal structure) in image A and image B and their correlation with the histopathological finding. The authors discuss differential diagnostically important differences of the echographic picture of myositis, endocrine myopathy and others. PMID- 11433594 TI - [Transmisssion electron microscopy of a submacular membrane imitating an intraocular tumor]. AB - The authors examined by transmission electron mycroscopy (TEM) a large submacular neovascular membrane which imitated clinically an intraocular tumour. TEM revealed in the sample fibroblast, pigmented retinal cells, parts of light sensitive cells, a haematoma and connective tissue. PMID- 11433595 TI - [Time factors and instrumental treatment of lacrimal duct obstruction in childhood]. AB - The authors evaluate the effectiveness of probing and rinsing of the lacrimal system in a group of 155 infants and young children (300 eyes) with congenital obstruction of the lacrimal pathways. The mean age during the first treatment was 3.7 months--range 21 days to 24 months. Patency of the lacrimal pathways was achieved in 83.49% of the group. In children treated before the 6th month the passage was restored in 91.30%. The effectiveness of procedures declines with advancing age. In the author's opinion it is best to make the operation between the 4th and 6th month of age. The operation under local anaesthesia ensures in the majority of children a rapid and permanent regression of complaints and a relatively small number of required repeated probings. At a later age the effectiveness of probes or rinsing declines and the solution is rather an indication for intubation of the lacrimal pathways or DCRS. One of the main still open questions for discussion is the optimal age for the operation. The author discusses also views of adverseries of probing and the problem of spontaneous canalization of the lacrimal pathways. PMID- 11433596 TI - [Diclof 0.1% eyedrops in pediatric ophthalmology]. AB - The authors analyze their two years experience with 208 patients hospitalized at the Clinic of Pediatric Ophthalmology treated by the locally administered antiprostaglandin diclophenac, 0.1% eye drops. The spectrum of the disease was wide and comprised different conditions after intraocular surgery, in particular of cataract, glaucoma and perforation injuries, Yag laser capsulotomy, and conservatively treated conditions, in particular inflammatory processes, uveitis and keratoconjunctivitis. Diclophenac was not combined with other non-steroid antiphlogistics but frequently it was adjuvant treatment of corticosteroids, antibiotics and mydriatics. The authors did not observe any side-effects of the preparation, even after long-term administration or after short-term administration to neonates. As to the intensity of effect, diclophenac was comparable with other prostaglandin inhibitors and in pediatric ophthalmology it is an effective and safe antiphlogistic agent. PMID- 11433597 TI - [Retinopathy in dermatomyositis]. AB - The authors describe a patient with acute onset dermatomyositis, "probable", according to criteria of Bohan and Peter, as well as the development of the disease in the course of 21 months. A special feature was the decline of visual acuity (practically to the level of blindness). The cause was severe retinopathy with dominance of macular oedema, "cotton wool" spots and haemorrhages, as apparent from photographs of the fundus and findings obtained by fluorescein angiography. The problem of initial treatment was resistance to megadoses of glucocorticoid (i.v. pulses). A favourable change occurred only after i.v. administration of polyvalent immunoglobulin in a total dose of 600 mg/kg body weight. Then the authors observed the unique phenomenon of regression of retinal oedema incl. the macula, of the haemorrhages and "cotton wool" spots. Consistent with it vision off the right eye improved to 1.0 and the left eye to 0.66 in the course of 11 months. Stabilized retinal neovascularizations persist. On account of the latter the authors treated the retina by laser panretinocoagulation. PMID- 11433598 TI - [Corneal topography in laser refractive surgery]. PMID- 11433599 TI - [George Friedrich Handel and his blindness. 23 February 1685 to 14 April 1759]. PMID- 11433600 TI - Alzheimer's disease, inflammation and non-steroidal anti-inflammatory drugs. AB - Alzheimer's disease (AD) is a degenerative disease of the brain, which causes dementia. The disease is characterised by three main pathogenic factors: senile plaques, neurofibrillary tangles and inflammation. the participation of the local inflammatory reaction is confirmed especially by the results of studies dealing with activated microglia, reactive astrocytes, complement system, cytokines, reactive mediators of oxygen and nitrogen (free radicals), all of which participate significantly in inflammatory processes. These inflammatory markers are locally produced by brain cells, and occur in close proximity of beta-amyloid and tau protein deposits. Moreover, some epidemiologic and pilot clinical studies have proven that long-term administration of anti-inflammatory drugs have a protective effect on the onset of AD. Out of them, non-steroidal anti inflammatory drugs (NSAIDs) are most extensively investigated medicaments. Despite some contradictory findings, the prevalent majority of these studies prove that long-term application of anti-inflammatory treatment can delay the onset, or at least slow down the progression of AD, namely in people between 65 and 75 years of age. The most appropriate prophylactic effect seems to be achieved by specific inhibitors of cyclooxygenase-2 (COX-2), namely celecoxib and rofecoxib. These preparations protect the gastrointestinal tract better than classical NSAIDs which inhibit both isoenzymes--COX-1 and COX-2. COX-2 is expressed in higher concentrations in the degenerating cells of the brain and this excessive expression can be decreased by selective inhibitors. The latter decrease also the excessive activation of some transcription factors (PPARgama and the nuclear factor kapa-B), which are responsible for the initiation of transcription of a number of pro-inflammatory genes. The selective inhibitors COX 2 can thereby have an anti-inflammatory effect operating on several levels. (Tab. 1, Fig. 1, Ref. 75.) PMID- 11433601 TI - [High-frequency pressure ventilation with high positive end-expiratory pressure in the treatment of respiratory failure in neonates]. AB - The authors present results of group of 13 neonates treated with high frequency positive pressure ventilation (HFPPV) with high positive end-expiratory pressure (PEEP) for severe respiratory failure. The ventilatory protocol was based on the following principles: a) higher mean airway pressure (MAP) to achieve adequate oxygenation, b) MAP titrated mainly with PEEP, c) fraction of inspired oxygen (FiO2) below 0.6, d) small tidal volumes 3-6 ml/kg, e) ventilatory rates to achieve normocapnia in newborns with persistent pulmonary hypertension and to allow permissive hypercapnia in others. During HFPPV, the maximum values for respiratory rate, PEEP, MAP and peak inspiratory pressures (PIP), the incidence of airleak and the need for inotropic support were recorded. The values for arterial partial pressure of oxygen (paO2), FiO2, paO2/FiO2 and MAP during conventional ventilation and 30 minutes after initiation of HFPPV were statistically analyzed. paO2 increased from 8.0 kPa (3.3-10.4) to 11.8 kPa (7.3 16.2, p < 0.001) and paO2 (torr)/FiO2 increased from 62.2 (24.7-101.2) to 157.5 (62.2-275.2, p < 0.001) 30 minutes after institution of HFPPV when MAP was increased from 11.8 cmH2O (9-13.8,) to 17.2 H2O (14.8-22.2) p < 0.001. This allowed turning down FiO2 from 1 (0.6-1) to 0.6 (0.4-1 p < 0.001). Maximal ventilatory rates used were in average 60/min (50-105), PEEP 8 cmH2O (6-10), PIP 30 cmH2O (26-45), MAP 18.8 cmH2O (14.8-22.2). Air leak did not occur in any patient. Catecholamines were used in 8 patients. The duration of ventilatory support lasted in average 6 days (2-18). All patients were successfully extubated. 5 of them required nasal continuous positive airway pressure (14 hours -7 days). (Tab. 3, Fig. 3, Ref. 19.) PMID- 11433602 TI - Our experience with the treatment of benign prostatic hyperplasia (BPH) with tamsulosin. AB - BACKGROUND: Alpha 1-blockers decrease the tension, ease the tonus of smooth muscles and thus alleviate the voiding and storage symptoms of the lower urogenital tract. OBJECTIVES: The goal of this study was to assess objectively the improvement of the voidings and storage difficulties in 72 randomly chosen patients suffering from Benign Prostatic Hyperplasia (BPH) prior to, and in the course of 15 months of treatment with this alpha 1-blocker in a dose of one capsule of tamsulosin daily after meal. METHODS: During the period of three years (1997-1999), we have treated and studied 72 patients suffering from lower urinary tract symptoms (LUTS). The age of the patients ranged from 55 to 80 years with the median age of 71 years. The patients were examined physically, by laboratory methods, by ultrasonography (USG) and by uroflowmetry. Anamnestic data were justified by the application of the IPSS questionnaire and blood pressure tests (BP). The eventual volume of residual urine, as well as the condition of the kidneys were checked by USG. The Qmax values were measured by uroflowmetry prior to, and during treatment (0-3 weeks--6 months). We have continued with this treatment even after 6 months, but we have discontinued the biochemical, haematological and uroflowmetrical assessments. On the other hand, we have continued with quarterly examinations of urine, digital rectal examinations (DRE), measuring of BP and IPSS evaluation. PSA was checked at least once a year. Our patients were checked in this way for 12 to 18 months (median of 15 months). One capsule of tamsulosin was administered daily after meal. RESULTS: The prostatoselective alpha 1-blocker tamsulosin is a well-tolerated medication applied in the treatment of BPH. We did not have to discontinue the treatment with any of the patients during the 15 months of assessment. It is equally well tolerated at a dose of one capsule of tamsulosin daily, administered after the morning or evening meal without any orthostatic defect symptomatology. The quality of life has improved by three symptomatic units; the blood pressure remained practically unchanged, the IPSS score for BPH decreased by 6.8 score and the Qmax was upgraded from 10.1 ml/s to 14.9 ml/s. This finding remained practically unchanged during the 15 months of assessment and later. CONCLUSION: In the course of the recent years, the conservative treatment of BPH by phytotherapeutics but mainly by alpha 1-blockers have resulted in a considerable reduction of transurethral resections of the prostate (TURP), and of the transvesical "open" prostatectomy (PE open). This reduction down to 50% can be observed nearly all over the world. The presence of alpha 1-a, alpha 1-b, alpha 1 d receptors in the lower urinary tract is a good prerequisite for successful treatment of voiding and storage symptoms. In the assessment of the effects of a 15-month continuous treatment by tamsulosin, we have registered a considerable improvement in the quality of life (QOL), an increase in the Qmax, and decrease in the IPSS score. The age of patients under or over 60 years as well as their weight did not play significant roles. We have not registered any interaction with other medications generally used by older patients. (Ref. 30.) PMID- 11433603 TI - [Is allergy a contraindication for tonsillectomy?]. AB - Many immunologists indicate that tonsillectomy (TE) performed in patients with allergy has an unfavourable impact on the development of allergy and asthma. This warning has significantly decreased the number of TE in children, and resulted in an increase in the occurrence of peritonsillar abscesses, and high incidence of obstructive breathing disturbances during sleep associated with developmental physical and mental consequences in children. The subjective of this study was to find out the influence of TE on the development of allergy in 29 children, in whom their allergy was confirmed 1-5 years after TE. The allergic disease has improved or disappeared in 24 children (82.8%), no changes were recorded in 4 children (13.8%), deterioration has occurred only in 1 child (3.4%) who developed asthma, while in common population, asthma occurs in 10% of children. The results of this study confirm that TE does not have an unfavourable impact on the development of allergy, and therefore allergy does not represent a contraindication for tonsillectomy. (Fig. 1, Ref. 22.) PMID- 11433604 TI - [Adrenal incidentalomas]. AB - The extensive use and progress in the improvement of imaging techniques brings about the growing incidence of incidentalomas, i.e. adrenal lesions accidentally revealed during the imaging of the abdominal cavity which was originally focused on the visualisation of organs other than adrenal glands. Despite the fact than the majority of incidentalomas (55-91%) represent benign adrenocortical adenomas, it is necessary to focus the diagnostic approach on the exclusion of malignity and hormonal activity. However, no common consent has been yet achieved in the management of incidentalomas. The authors review the literature data on the prevalence, diagnosis (namely hormonal assessments as well as imaging methods) and the current suggestion of the necessity of further investigation and treatment of adrenal incidentalomas. (Tab. 1, Fig. 5, Ref. 64.) PMID- 11433605 TI - Implementation guidelines for effective management of hospital accreditation. AB - The introduction of accreditation in the health sector assumes that the management of health care facilities is ready for such a task. The preparation of management should be helped by guidelines, which will make hospital managers familiar with individual steps of preparation for successful accreditation. Overall, they should help hospital management to avoid unnecessary mistakes and losses. This paper involves the development of a set of guidelines intended to help hospital management prepare for accreditation. The implementation guidelines were prepared according to a summary of suggestions taken from international literature and the pilot study of accreditation in Slovakia. The experience of countries with a long history of accreditation is utilised. These guidelines divide the preparatory phase into ten steps. (Fig. 2, ref. 13.) PMID- 11433606 TI - Contribution to systematic education of quality management in Slovak health care. AB - THE AIM: Of the study was to contribute to quality improvement initiatives in Slovak health services through systematic approach to the education and training in quality management (QM). Consequently, the main objectives were to analyse the content of the education in QM abroad, to conduct an audit of perceived training needs in Slovakia, and to propose the design of QM training programme to be applied within CME scheme based on the study results. Triangular method in the design of the study was implemented. Review of relevant information, data from the questionnaire and semi-structured interview in the sample of 67 Slovak trainees from Health Management School and School of Public Health--were adopted in complementary fashion. BETWEEN FINDINGS: Highlighted in the survey are positive attitudes to training in quality management documented by the median score higher than 6 in all tested areas, on scale 0-10. No significant differences in profession groups as physicians, nurses, HC managers or among training institutions involved were displayed. However, potential obstacles were identified in deeper study using interviews. The absence of knowledge and skills in management in general and in quality management approaches especially are observed. Typically, the role of strategic planning is undermined. The large scale of quality management approaches is converted to problems of accreditation. Barriers to participative culture, innovation, devolution of accountability, resistance to change and to team based management are authentic findings as well. CONCLUSIONS: Drawn from the study were related to: fostering managers- "transformational leaders" for locally driven decision making in health care policy and practice; need of training activities for the continuing education in quality with respect to specific target groups interests and their level of knowledge in management; content of training oriented towards combination of rational utilization of information, critical analytical skills and planning for quality with human resource development-interpersonal skills, team building (soft skills), not just reduction of quality management tools to hard techniques (statistics, ISO norms); methods of education, where the usage of experiential learning methods, participative training inclusive action learning is highlighted; team training complemented with individual professional development support inclusive a coaching and mentoring scheme. AS IMPLICATIONS: Four types of CME training: Basic Module QM, Training for QM teams, Training Trainers Scheme and Guiding through Accreditation and Quality Award were proposed. (Tab. 9, Ref. 38.) PMID- 11433607 TI - Anatomic nomenclature by Vesalius. AB - Our paper deals with the situation of medieval anatomy language in the pre Vesalian period. It also explains the necessity of terminology reform undertaken by Vesalius in his De humani corporis fabrica. It describes formation principles of the scientific language of anatomy based on pure classical Latin while eliminating Arabic and Greek elements. Emphasis is laid on Vesalius' effort to unify terms as far as their meaning is concerned, to record lexical items and to create a permanent nomenclature in order to eliminate discrepancies in this field of communication. Our paper contains a lot of information on terminology demonstrating Vesalius' language reform and reminding us of his great achievement, for which he is considered a forerunner of anatomy nomenclature codification. (Ref. 5.) PMID- 11433608 TI - Synonymy of medical terminology from the point of view of comparative linguistics. AB - The aim of the paper is to look at the medical terminology in three different languages--English, Russian and Slovak and to estimate the trends of synonym formation as well as their emphasizes role in communication. The author that the problem of synonymy is peculiar especially to clinical medicine terminology, i.e. it relates only to a small extent anatomical nomenclature. According to motivative signs of terms, synonyms are divided into 2 groups: equivalent and interpretative synonyms. In keeping with criteria determined by basic characteristics of a term in medical terminology there are often preferred international expressions. In recent years a great number of English expressions have entered into medical terminology due to the fact that English had became a language of international communication. From the practical point view undoubtedly, it is a positive tendency, because it facilitates international communication. In this respect Russian orthography is considered to be an exception, since Cyrillic alphabet often makes international communication more complicated. (Ref. 10.). PMID- 11433609 TI - [A model of development of the number of physicians in Slovakia]. AB - The aim of this study was to work out a mathematical model of the anticipated development of the number of physicians in Slovakia during the period 2000-2025. The study is based on the current state of professionally active physicians employed by state and private health care organisations, their age structure, and the assumed number number of graduates of medical universities. This study is the first work trying to produce a model of this problem. PMID- 11433610 TI - [Measurement of HER-2/neu in breast cancer: which methodologic approach?]. AB - HER-2/neu (c-erbB2) is overexpressed or amplified in 15%-35% of breast cancers. HER-2 testing has become of paramount importance as it represents a key therapeutic, prognostic and predictive parameter. HER-2 is associated with ER/PgR negativity, high histologic grade, high proliferative index, and increased number of metastatic lymph nodes. In N+ patients, HER-2 represents a negative prognostic factor. HER-2 also represents a predictive factor. In fact, HER-2+ patients appear to be resistant to the CMF regimen and comparatively more sensitive to anthracyclins such as Herceptin. HER-2 testing is currently performed utilizing two main methods: gene amplification is usually determined by fluorescence in situ hybridization (FISH), whereas protein overexpression is determined by immunohistochemistry (IHC). The comparison between FISH and IHC assays demonstrates a considerably high degree of concordance (around 90%). When dealing with cases scored as 3+ (according to the FDA licensed scoring method), such a concordance approaches 100%. In consideration of greater technical simplicity as well as of cost-effectiveness, IHC represents the ideal screening method for HER 2 testing. FISH analysis remains valid for HER-2 evaluation in those cases in which IHC fails to provide unequivocal data. PMID- 11433611 TI - Relevance of the rates of PCNA, Ki-67 and p53 expression according to the epithelial compartment in cervical lesions. AB - In order to assess further biological evidence for similarities among the "diagnostic classes" of cervical lesions, which are now a matter of international discussion in the search for a uniform classification, the purpose of this study was to characterize the immunoexpression of cell proliferation markers (proliferating cell nuclear antigen, PCNA and Ki-67) and protein p53. Each marker was individually quantified in basal, intermediate, and superficial epithelial compartments presenting chronic cervicitis (CC) accompanied by the cytopathic effects of infection by human papillomavirus (CCHPV) or not (CC), as well as in cervical intraepithelial neoplasia (CIN) grades I, II, and III. A total of 100 patients were evaluated and the positive nuclei were counted separately, including all extensions of the available epithelium. The percentage of PCNA- and Ki-67-positive cells increased with increasing grade of the cervical lesions, although PCNA immunoreactivity was always greater than the immunoreactivity observed with Ki-67 antigen. The immunoexpression of p53 protein was found to be weak, with no remarkable behavior in any specific "diagnostic class". The differences in cell proliferation markers found herein further emphasize the progressive loss of epithelial layer organization in the course of the development of preneoplastic changes in cervical squamous epithelium. Furthermore, difficulties in morphologically distinguishing "borderline lesions" persist when cell cycle markers are studied, further supporting the suggestion to consider the lesions of CCHPV and CIN I together as only one diagnostic class. Conversely, the different immune profile found between CIN II and III further supports the validity of the subdivision of CIN into three groups. PMID- 11433612 TI - [The role of the pathological anatomy museum: past glory, present crisis, and future prospects]. AB - Scientific Museum and in particular pathology Museum were in the past an important landmark for Pathology and in general for Medicine. At present they are in a critical stage due to the technological progress, because they are considered by now obsolete compared to other didactic supports, so as for example CD-ROM or virtual presentations. In opposite these old collections are still very precious for Medicine, because they show old diseases, now disappeared or very rare, thanks to the therapeutic and diagnostic progresses. Furthermore they are very useful for teaching of Pathology to medical students and also to laboratory's technicians. The specimens may be used also for organisation of exhibitions, in graduate courses or medical updating courses, for a better popularisation of Anatomy or of some diseases of large environmental or social impact. For all these reasons we propose the creation of renewed museum structures and a protocol for the restoration of the specimens, which are often damaged owing to time and negligence. PMID- 11433613 TI - Mapping a method for systematically reviewing the medical literature: a helpful checklist postmortem protocol of human immunodeficiency virus (HIV)-related pathology in childhood. AB - INTRODUCTION: Since the first description of two children affected with human immunodeficiency virus (HIV), various diagnostic procedures have been established. However, the morphologic study of biopsy and autopsy material from children with acquired immunodeficiency syndrome (AIDS) is still of fundamental importance. The morphology has contributed remarkably to the progress in understanding the pathogenesis of the primary tissue lesions and of the sequelae directly or indirectly associated with HIV infection. The aim of this study was to evaluate the HIV pathology in pediatric AIDS (PAIDS) through a systematic review of the English-language literature and to draw up a practical checklist protocol for the postmortem. MATERIALS AND METHODS: Information on HIV pathology in childhood was retrieved from a MEDLINE search (January 1994-January 2001) of the original reports and bibliographic article reviews published in English. Citations from papers retrieved were screened and retrieved papers were evaluated. RESULTS: Based on the screened data, we propose a practical, organ oriented checklist protocol for the postmortem according to the HIV pathogenesis. CONCLUSIONS: Evidence-based medicine is a paradigm now exerting increasing influence in related fields such as surgery, general practice, psychiatry, and pathology. This article is a summary of the literature on PAIDS pathology. The protocol that we propose is particularly useful for pediatric pathology programs and for electronic data processing. PMID- 11433614 TI - [Endocrine differentiation in basocellular carcinoma]. AB - Three (propositus) cases of basal cell carcinoma (BCC) showing endocrine differentiation at the immunohistochemical level were studied using reverse transcriptase-polymerase chain reaction (RT-PCR) to determine the presence of mRNA of chromogranin A. Moreover, 20 (consecutive) cases of BCC were studied with immunohistochemistry alone using chromogranin A, synaptophysin, S100 protein, cytokeratin 20, and neuron-specific enolase antibodies (NSE). The three propositus cases of BCC showed positive results when RT-PCR for mRNA of chromogranin A was performed. Eleven out of 20 consecutive cases of BCC were focally positive for chromogranin A antibody. These results confirm the presence of endocrine differentiation in BCC, demonstrated both with immunohistochemistry and with RT-PCR. PMID- 11433615 TI - [Visceral pleural lipoma: description of a autopsy case]. AB - Pleural lipomas are rare tumours, usually arising from the parietal pleura, which are often asymptomatic and observed incidentally. We describe the clinicopathological features of a case of lipoma arising from the visceral pleura, accidentally discovered in a 69-year-old ma, at autopsy. A review of the literature concerning pleural lipomas is also presented. PMID- 11433616 TI - [Combined neoplasia of the lung: description of a case of adenocarcinoma mixed with typical carcinoid]. AB - OBJECTIVE: To report about a lung tumor that was a combination of typical carcinoid and adenocarcinoma. RESULTS AND CONCLUSIONS: The patient, a 71-year-old male, presented with a 2.5-cm pulmonary nodule that, microscopically, was a combination of an adenocarcinoma (tubular with clear cell features and bronchioloalveolar) and a typical carcinoid. Immunohistochemically, both components were positive for cytokeratin, but only the carcinoid component was positive for chromogranin and synaptophysin. In the range of neuroendocrine tumors of the lung, a combination with other histological types of carcinoma (squamous, adeno, large cell and pleomorphic) can be found with both small cell carcinoma and large cell neuroendocrine carcinoma, but is very rare with typical and atypical carcinoids. PMID- 11433617 TI - [Lymphoepithelioma-like carcinoma of the esophagus: description of a case]. AB - The authors describe a case of lymphoepithelioma-like carcinoma (LELC) of the esophagus occurring in a 77-year-old man. To date, 13 cases have been described in the literature, all in natives of Japan. Histological features, immunohistochemical findings and differential diagnosis are discussed. In our case epithelial neoplastic cells were completely negative for the EBV genome. PMID- 11433618 TI - Assessment of radical prostatectomy specimens and diagnostic reporting of pathological findings. AB - Proper examination of radical prostatectomy (RP) specimens from patients with prostate cancer (PCa) is critical for determining the need for adjuvant treatment and for predicting the outcome. The first step in the examination is represented by the assessment of the specimens. This includes proper fixation in formalin and whole-mount sectioning. The second step is related to the diagnostic reporting of pathological findings. This includes the evaluation of tissue features such as Gleason grade, stage and margin status. These three parameters, together with serum prostate-specific antigen (PSA) determination, are predictive factors whose prognostic role has been demonstrated in several studies. PMID- 11433619 TI - [Personnel training for a population screening program for cervical carcinoma]. PMID- 11433620 TI - Improving the value of fine needle aspiration biopsy of the prostate. PMID- 11433621 TI - [Some investigations by Giuseppe Bini: examples of cultural journeys through pathological anatomy and history]. PMID- 11433623 TI - [About autopsies]. PMID- 11433624 TI - [Reporting melanocytic lesions: the form, substance, and the void]. PMID- 11433625 TI - Using outcome evaluations to assess interdisciplinary acute and chronic pain programs. AB - BACKGROUND: Outcome evaluations can be useful for reassuring patients that their time and effort are well spent on treatment and for providing staff with confidence in their treatment provision. Outcome evaluations were carried out in 1997 and 1999 to assess two initiatives for the treatment of patients within the Regina Health District (RHD), Saskatchewan, Canada-the Chronic Pain Team Evaluation and Management (CP TEAM) Service and the Acute Injury Management (AIM) Program. STUDY 1: The CP Team Service was an individualized interdisciplinary treatment program intended to serve patients with chronic pain. Of 47 eligible chronic pain patients, 36 participants completed treatment within 15 months, and 27 completed follow-up questionnaires on discharge; the comparison group was composed of the remaining 11 nontreated participants, 8 of whom responded to follow-up questionnaires. Only patients in the treatment group reported a decrease in pain and a decrease in interference in activities. Overall, satisfaction with treatment received was high. STUDY 2: The AIM Program was intended to aid employees with acute musculoskeletal injuries who could not perform regular job duties. Following injury, employees were contacted by an occupational health nurse and informed of the program. If the employee was interested, the AIM coordinator carried out an assessment and designed an individualized treatment package, including physical therapy, exercise therapy, and/or occupational therapy. Treatment was expected to continue until the employee returned to normal duties. Of 72 eligible employees, 43 employees participated and 29 opted not to participate-15 of whom agreed to participate as control participants for this study. Of the employees receiving AIM, 20 agreed to participate in this study. Of the employees not participating in AIM, 15 agreed to participate as control participants. Although pain and interference were greater among AIM participants before commencing treatment, by the end of treatment participants had improved more in level of pain severity and interference compared with controls. All participants reported improved job function over time. AIM participants, however, reported lower job function before treatment than control participants and similar job function after treatment- and indicated they were highly satisfied with the service they received. SUMMARY AND CONCLUSIONS: Evaluations are not only used to assess program outcomes but are an important aspect of program validation and development. Data collection was incorporated, as much as possible, into routine treatment protocols and staff focused on obtaining essential information regarding patient outcomes rather than the whole scope of information. PMID- 11433626 TI - Patient satisfaction measurement strategies: a comparison of phone and mail methods. AB - BACKGROUND: In 1998 the BJC Health System (St Louis) made the decision to migrate its patient satisfaction measurement system from a mail-out/mail-back method to a phone interview method. Out of concern that results obtained by phone would not be comparable with the 4 years of mail-based data, a controlled study was undertaken to directly compare mail and phone responses and to evaluate response rates, patient sample demographics, and patient satisfaction ratings. METHODS: Mail and phone responses obtained from parallel random samples selected from inpatient, outpatient test/treatment, outpatient surgery, and emergency service patient populations were compared. Patients were randomly selected to receive a standardized satisfaction survey by either phone or mail 10 to 14 days postdischarge. RESULTS: Significantly higher response rates were obtained by telephone then via the mail-based method for all four samples. After adjusting for demographic differences, numerous significant differences in mean scores as well as percentages of excellent and fair or poor responses were observed, and more positive ratings were obtained by phone. DISCUSSION: Crude comparisons of satisfaction scores between organizations using phone and mail-based responses may lead to erroneous conclusions about consumer-perceived quality. Organizations that use mixed-mode surveys should conduct careful side-by-side studies of the methods used on the survey of interest and then establish a correction formula to adjust the results for the measurement biases. PMID- 11433627 TI - Using "good and bad surprises" to guide improvement efforts: insights from a memory assessment clinic. AB - BACKGROUND: Continuous quality improvement requires an understanding of customer expectations, yet often little is known about what patients and their families expect from specialized diagnostic services. A project was conducted in 2000 at two memory assessment clinics affiliated with the University of Kentucky's Alzheimer's Disease Research Center (ADRC) to use open-ended "good and bad surprises" to learn what families expect from a memory assessment clinic and to demonstrate how "surprises" might be used to identify customer expectations. METHODS: A satisfaction survey was sent to persons who had had a role in the decision to seek a memory assessment for a family member within the previous 4 years. The responses to an open-ended question, which formed the basis for this study, were coded for process and outcome expectations. RESULTS: Of the 528 (71.7% response rate) surveys returned, 390 (73.8%) included a response to the open-ended question designed to elicit good and bad surprises. Approximately one half of the respondents had accompanied a parent and one-half of the respondents had accompanied a spouse to the memory assessment clinics. The diagnosis received and the provision of information caused the most surprises. Provision of information contributed to both good and bad surprises. Emotional support, provider attitudes, treatment availability, and the quality of the exam were additional causes of good surprises. The diagnosis received and family reactions caused bad surprises. The majority of the good surprises related to process expectations, and the majority of the bad surprises related to outcome expectations. DISCUSSION: By eliciting information about good and bad surprises, organizations can identify their customers' expectations and identify areas for improvements. Assessment clinics should undertake improvement efforts designed to prepare customers for the diagnosis received and provide their customers with tailored information. PMID- 11433628 TI - Integrating baseline health status data collection into the process of care. AB - BACKGROUND: Health status data are an increasingly important component of outcomes assessment and can be used to facilitate quality assessment and improvement efforts. An enormous challenge to the use of health status data among hospitalized patients, however, is collecting baseline data at the time of treatment, an essential component for risk-adjusting subsequent outcomes. The Mid America Heart Institute of Saint Luke's Hospital (Kansas City, Mo), attempted to integrate the collection of health status assessments within the process of performing coronary revascularization. THE DATA COLLECTION STRATEGY: The data collection strategy was developed for each admission portalelective outpatients (admissions for same-day procedures), inpatients, and emergent cases. Health status data were collected on all patients with coronary artery disease who were receiving a percutaneous coronary intervention or coronary artery bypass graft with no disruption to physician scheduling or nursing staff. RESULTS: In general, patients were agreeable to completing the health status survey. Despite initial efforts to educate the hospital staff about the goal and purpose of health status assessment, staff members who were unaware of the uses of these data seemed to minimize their value. Providing examples of how to use these data relative to the staff member's specific occupational role facilitated buy-in for this project. EPILOGUE: After the pilot study, which lasted until June 1999, data were continually collected for 18 months, through August 2000, even with the cessation of external grant funding for this project. Baseline data collection finally stopped, primarily because of a failure to accommodate data collection into the routine flow of patient care by existing nursing staff. PMID- 11433629 TI - Is participation in cardiac rehabilitation programs associated with better quality of life and return to work after coronary artery bypass operations? The Israeli CABG Study. AB - OBJECTIVE: To explore the putative effect of cardiac rehabilitation programs on the 'health-related quality of life' and 'return to work' in pre-retirement patients one year after coronary artery bypass grafting. METHODS: Of the 2,085 patients aged 45-64 who survived one year after CABG and were Israeli residents, 145 (6.9%) had participated in rehabilitation programs. Of these, 124 (83%) who answered QOL questionnaires were individually matched with 248 controls by gender, age within 5 years, and the time the questionnaire was answered. All patients had full clinical follow-up including a pre-operative interview. The Short Form-36 QOL questionnaire as well as a specific questionnaire were mailed to surviving patients one year after surgery. Study outcomes included the scores on eight scales and two summary components of the SF-36, as well as 'return to work' and 'satisfaction with medical services' from the specific questionnaire. Analysis was done for matched samples. RESULTS: Cardiac rehabilitation participants had significantly higher SF-36 scores in general health, physical functioning, and social functioning. They had borderline significant higher scores in the physical summary component of the SF-36. The specific questionnaire revealed significantly better overall functioning, higher satisfaction with medical care, and higher rate of return to work. While participants in cardiac rehabilitation and their controls were similar in their socio-demographic and clinical profiles, participating patients tended to be more physically active and more fully employed than their controls. CONCLUSIONS: Rehabilitation participants had a self-perception of better HRQOL, most significantly in social functioning. Our findings of more frequent return to work and higher satisfaction with medical care should induce a policy to encourage participation in cardiac rehabilitation programs after CABG. PMID- 11433630 TI - Prevalence of selected chronic diseases in Israel. AB - BACKGROUND: Knowledge of the prevalence of chronic disease in the population is essential for health planners and providers. OBJECTIVES: To present the results of a concentrated effort by the largest health maintenance organization in Israel (Clalit Health Services) in order to develop a comprehensive register of chronic diseases. METHODS: In 1998, all 2,704 primary care physicians in Israel's largest health provider were requested to report on all patients with selected chronic diseases. In addition, all the filled prescriptions for medications relevant to the investigated diseases and all relevant hospitalization events were added to the database. Prevalence rates were calculated based on the reporting practices only (1,653 physicians responsible for a total of 1,409,725 adults). RESULTS: Hypertension (10.2/100), diabetes (6.1/100), hyperlipidemia (5.7/100), peptic ulcer (4.7/100) and ischemic heart disease (4.3/100) were the most prevalent. Females had significantly higher rates of hypothyroidism, psychoses, neuroses and malignancies, and lower rates of ischemic heart disease, chronic obstructive pulmonary disease, heart failure and asthma. Arabs had higher rates of diabetes mellitus and lower rates of ischemic heart disease, hypertension and hyperlipidemia than Jews. About 20% of the adult population had one or more of the selected chronic diseases. CONCLUSIONS: Differences in rates noted between physicians, not explainable by population characteristics, may reflect differences in the quality and delivery of health services. Rate differences between demographic subgroups call for further studies on the etiology, susceptibility and natural history of these diseases. PMID- 11433631 TI - Thyroglossal duct cyst: is thyroid scanning necessary in the preoperative evaluation? AB - BACKGROUND: Evaluation of thyroglossal duct cysts before surgical excision traditionally includes hormonal evaluation, ultrasound of the neck, and thyroid scanning. OBJECTIVE: To evaluate the need for thyroid scanning in cases of thyroglossal duct cysts scheduled for operation. METHODS: A retrospective chart review of 100 cases of thyroglossal duct cyst between 1988 and 1995 was carried out. RESULTS: No cases of ectopic thyroid were detected. CONCLUSION: Our goal was to document the presence of normal thyroid tissue by non-invasive tools such as ultrasound, rather than to rule out the existence of ectopic thyroid tissue by radionuclide scanning. Radionuclide scanning is justified in cases of lingual thyroid and where a normally located thyroid gland cannot be detected. PMID- 11433632 TI - Ultrasonography as a diagnostic modality and therapeutic adjuvant in the management of soft tissue foreign bodies in the lower extremities. AB - BACKGROUND: Foreign bodies are sometimes overlooked in the initial evaluation of soft tissue wounds in the emergency room setting. The physical examination identifies foreign bodies that are superficial enough to be seen or palpated, while radiographs reveal those that are radio-opaque. If these two criteria are not met, however, the foreign body may remain undetected. These patients present later with long-standing pain in the area of penetration sometimes associated with localized tenderness. OBJECTIVES: To assess the role of ultrasonography in the diagnosis and management of patients with a suspected retained foreign body. METHODS: Ultrasound was used in 21 patients with suspected retained foreign bodies and the diagnosis was positive in 19. Fifteen underwent a surgical exploration in which the ultrasound was used as an adjunctive modality either pre or intraoperatively to assist in the localization of the foreign body. RESULTS: All procedures were successful. No postoperative complications were recorded at an average follow-up of 2 years. Three patients gradually became asymptomatic and were left untreated. One patient was lost to follow-up. CONCLUSION: Sonography is an extremely effective tool for the late diagnosis of retained foreign bodies in the soft tissues. We suggest that its availability in the emergency room may decrease the rate of misdiagnosis and avoid these unfortunate cases, although this remains to be proven. PMID- 11433633 TI - Computerized tomography findings of abdominal tuberculosis: report of 19 cases. AB - BACKGROUND: Abdominal tuberculosis usually presents with general symptoms and obscure abdominal complaints for which computerized tomography is often the first imaging study. OBJECTIVE: To evaluate the CT findings of abdominal tuberculosis. METHODS: The CT scans of 19 patients (10 men and 9 women aged 20-85 years) with proven abdominal tuberculosis were retrospectively reviewed to define the location and extent of the disease. The patients were referred for the study mainly with general systemic symptoms. Additional abdominal complaints were present in four, including acute abdomen in one. Two had symptoms deriving from the urinary tract. Nine patients had recently arrived from high prevalence countries; five of them and two others were positive for human immunodeficiency virus. Three patients had a family history of tuberculosis; one had previously been treated for tuberculosis and four others had an underlying chronic disease. The diagnosis of tuberculosis was established by standard microbiological and histological techniques. RESULTS: We divided the disease manifestations into intraperitoneal (n = 13) and genitourinary involvement (n = 6). Peritoneal tuberculosis was fairly common, characterized by ascites, omental and mesenteric infiltration, and smooth thickening of the parietal peritoneum. One oncology patient had a false positive Tc-99m CEA isotope scanning, suggesting tumor recurrence. Genitourinary disease manifested mainly as hydronephrosis and calcifications. Three patients had pulmonary tuberculosis as well. CONCLUSION: The CT findings of abdominal tuberculosis may mimic various diseases, mainly diffuse peritoneal malignancy. We emphasize the need to consider tuberculosis in the differential diagnosis in patients with obscure abdominal symptoms, especially with multi-organ involvement. A high degree of clinical suspicion and familiarity with the abdominal CT manifestations allow early diagnosis of this treatable disease. PMID- 11433634 TI - Vitamin D deficiency and insufficiency in Orthodox and non-Orthodox Jewish mothers in Israel. AB - BACKGROUND: The modest clothing that Orthodox Jewish women wear exposes very little of their skin to sunlight. Under these conditions they may develop vitamin D deficiency, even in sunny Israel. OBJECTIVES: To determine and compare the vitamin D nutritional status in Jewish orthodox mothers to that of non-orthodox mothers who live in the same metropolitan area in Israel. METHODS: 25 Hydroxyvitamin D was measured by competitive protein-binding radioassay in the sera of 341 Jewish Israeli mothers (156 orthodox and 185 non-orthodox). The sera were obtained 48-72 hours after childbirth during the late summer of 1998 and the spring of 1999. RESULTS: The mean (SD) serum concentration of 25-OHD was significantly (P < 0.002) lower (13.5 +/- 7.5 ng/ml) in the orthodox than in the non-orthodox mothers (18.6 +/- 9.6 ng/ml). Vitamin D deficiency (< 5 ng/ml) and insufficiency (< 10 ng/ml) were more common in the orthodox mothers (5.1% and 32.7% respectively) than in the non-orthodox mothers (2.7% and 13%, respectively). In subgroups of mothers supplemented with 400 units of vitamin D daily during pregnancy, vitamin D deficiency and insufficiency were less common (2.2% and 13%, respectively) in orthodox and non-orthodox mothers (0% and 8.1%, respectively). Vitamin D insufficiency was more common in the winter than in the summer only among non-orthodox mothers. CONCLUSIONS: The high prevalence of vitamin D deficiency and insufficiency in Israeli mothers raises the question whether vitamin D supplements should be given to pregnant women in Israel, at least to orthodox mothers. PMID- 11433635 TI - Newspaper advertising by health maintenance organizations during the reform of healthcare services in Israel. AB - BACKGROUND: On 1 January 1995 a new mandatory National Health Insurance Law was enacted in Israel. The new law fostered competition among the four major Israeli healthcare providers (HMOs or sick funds) already operating in the market due to the possibility that an unlimited number of patients and the relative budget share would shift among the HMOs. This led them to launch advertising campaigns to attract new members. OBJECTIVES: To examine newspaper advertising activities during the early stages of healthcare market reform in Israel. METHODS: Advertising efforts were reviewed during a study period of 24 months (July 1994 to June 1996). Advertisements were analyzed in terms of marketing strategy, costs and quality of information. RESULTS: During the study period 412 newspaper advertisements were collected. The total advertising costs by all HMOs was approximately US$4 million in 1996 prices. Differences were found in marketing strategy, relative advertising costs, contents and priorities among the HMOs. CONCLUSIONS: The content of HMOs' newspaper advertising was consistent with their marketing strategy. The messages met the criteria of persuasive advertising in that they cultivated interest in the HMOs but did not provide meaningful information about them. Future developments in this area should include consensus guidelines for advertising activities of HMOs in Israel, instruction concerning the content of messages, and standardization of criteria to report on HMO performance. PMID- 11433636 TI - Cardiac rehabilitation after coronary artery bypass graft surgery: urgently needed! PMID- 11433637 TI - Mixed cryoglobulinemia and hepatitis C virus association: ten years later. AB - Mixed cryoglobulinemia is a systemic vasculitis described as a triad characterized by purpura, weakness and arthralgias. Since the first description of the disease in 1964 by Meltzer and Franklin our understanding of its pathogenesis has increased considerably. The striking association of the disease with hepatitis C virus infection was initially noted in 1990. Since then, the disease has gained growing attention among investigators involved in the study of autoimmune systemic disorders because it represents one of the most intriguing models of autoimmunity triggered by a virus. Nonetheless, a number of answered questions still remain to be resolved and are reviewed in this article. PMID- 11433638 TI - Heredity in Parkinson's disease: new findings. AB - Multiple factors have been hypothesized over the last century to be causative or contributory for Parkinson's disease. Hereditary factors have recently emerged as a major focus of Parkinson's disease research. Until recently most of the research on the etiology of Parkinson's disease concentrated on environmental factors, and the possibility that genetic factors contribute significantly to the pathogenesis of Parkinson's disease has been neglected. However, it has become increasingly apparent that even in sporadic cases, the disease most likely reflects a combination of genetic susceptibility and an unknown environmental insult. Moreover, the identification of genes and proteins that may cause hereditary parkinsonism substantially contributes to our ability to understand the pathogenesis of Parkinson's disease and may help in the early identification of the disease and its treatment. The discovery of alpha-synuclein mutations in families with autosomal dominant Parkinson's disease sheds light on its role in sporadic Parkinson's disease. It seems that this protein tends to aggregate when the cellular milieu is altered [14-16]. The question as to the exact changes that cause its deposition remains open. One of the major possibilities is oxidative stress [16]. The role of these aggregates in neuronal cell death is also still unclear. Transgenic mice expressing wild-type human alpha-synuclein developed progressive accumulation of alpha-synuclein and ubiquitin-immunoreactive inclusions in neurons in the neocortex, hippocampus and the substantia nigra. These alterations were associated with loss of dopaminergic terminals and motor impairments [24]. This finding suggests that accumulation of alpha-synuclein may play a causal role in sporadic Parkinson's disease as well. The parkin protein seems to be a crucial survival factor for nigral neurons [15]. The parkin protein is related to the ubiquitin pathway, which is important in the elimination of damaged proteins. Ubiquitin-mediated degradation of proteins plays a central role in the control of numerous processes, including signal transduction, receptor and transcriptional regulations, programmed cell death, and breakdown of abnormal proteins that may interfere with normal cell functions. Further studies on the function of Parkin protein and its relation to the ubiquitin pathway could elucidate at least one of the molecular mechanisms of nigral neuronal death. A mutation in the ubiquitin carboxy-teminal hydrolase L1 gene also implies the importance of the ubiquitin pathway in Parkinson's disease. Abnormal tau protein was found to be the cause of familial frontotemporal dementia and parkinsonism. It tends to form filamentous structures, which may lead to neuronal death. Elucidation of the molecular mechanism of neuronal death in this disease may contribute to our understanding of sporadic diseases with tau accumulation, such as corticobasal degeneration, progressive supranuclear palsy, Pick's disease, Alzheimer's disease and possibly also the pathogenesis of Parkinson's disease. Other genetic loci have been identified by linkage analysis of patients with familial parkinsonism. These loci conceal other genes and proteins that may be pivotal factors in the pathogenesis of Parkinson's disease. The discovery of genetic mutations in patients with parkinsonism may offer us new insights into the understanding of the pathways leading to neuronal death and development of Parkinson's disease. It may also help in the early identification of susceptible people to this disease and possibly in developing new treatment strategies. PMID- 11433639 TI - New approaches in the diagnosis of sepsis. AB - In diagnosing sepsis the rapid identification of bacteremia at an early stage of the disease is critical for a favorable outcome. Furthermore, it is important that exact information on the stage of the disease be obtained rapidly in order to choose and initiate the appropriate therapy. In recent years many new techniques have been added to the diagnostic tools. In this review we will focus on three new methods for the early diagnosis of sepsis. These are: polymerase chain reaction, which offers the possibility to attain detailed information about the involved bacterial (or viral) species, and the laboratory markers procalcitonin and hypophosphatemia, which are indicators of the presence of infection with gram-negative bacteria. The approaches reviewed here were developed to expedite the diagnosis of especially early sepsis and might be a further step towards the improvement of therapy for sepsis. PMID- 11433640 TI - Ocular blood flow measurements and their importance in glaucoma and age-related macular degeneration. AB - This survey of methods for assessing ocular hemodynamics in glaucoma and age related macular degeneration is not complete, but it does cover those likely to be encountered in the literature. A fundamental problem in getting to grips with the ocular blood flow literature is the difficulty in comparing the results of similar studies employing different assessment techniques. As evident from the discussion above, each technique evaluates a portion of the ocular circulation in a distinct way. Some of the methods overlap with regard to the tissues that can be used for examination, while others are directed at entirely different parts of the ocular vasculature. Despite these difficulties, hemodynamic studies of glaucoma and AMD are likely to grow in importance. On the basis of accumulating epidemiological and clinical evidence, it is becoming apparent that intraocular pressure is not the sole etiological factor in glaucoma, and retinal pigment epithelium senescence is not the sole etiological factor in AMD. Circumstantial evidence of vascular involvement in glaucoma and AMD has now been bolstered by experimental evidence. If the current pace of refinement of newly established technologies for evaluating ocular blood flow is maintained, they will soon be ready for deployment in the clinic. The only problem is the availability of expensive instruments and trained personnel. The ultimate beneficiaries of work in this area will not be researchers, but patients. PMID- 11433641 TI - Liver cirrhosis and the hyperglycemic hyperosmolar non-ketotic state. PMID- 11433642 TI - Post-kala-azar dermal leishmaniasis manifesting after initiation of highly active anti-retroviral therapy in a patient with human immunodeficiency virus infection. PMID- 11433643 TI - Successful use of high dose intravenous immunoglobulin in rapidly progressive crescentic glomerulonephritis with vasculitis. PMID- 11433644 TI - The use of 75Se-Selenocholestrol SPECT in the localization of steroid-secreting tumor. PMID- 11433645 TI - Superior vena cava syndrome diagnosed by transesophageal echocardiography. PMID- 11433646 TI - A deadly venopuncture. PMID- 11433647 TI - Filter placement in duplicated inferior vena cava. PMID- 11433648 TI - Medical ethics committees in Israel: implementing the Israel Patient Rights Act and terminating life-sustaining treatment. PMID- 11433649 TI - Horizon scanning in medical education: 2020 vision. PMID- 11433650 TI - Acute mechanical obstruction of the colon in scleroderma. PMID- 11433651 TI - Ambulatory utilization of primary care. PMID- 11433652 TI - Echinococcosis in pregnancy. PMID- 11433653 TI - Cerebral salt wasting syndrome or secondary hypoadrenalism? PMID- 11433654 TI - More on hospital utilization and inpatient care. PMID- 11433655 TI - Sarcoidosis and Sjogren's syndrome. PMID- 11433656 TI - Myositis accompanying Rickettsia conorii infection. PMID- 11433657 TI - Drug-induced QT prolongation. PMID- 11433659 TI - [Increasing importance of cytological diagnostics in the Netherlands]. AB - Cytological diagnosis has important practical clinical value, as shown by the observation that 54% of all diagnostic activities in pathology laboratories consist of cytological investigations. New impulses are, however, required to ensure ongoing developments. For example, from a scientific point of view, there appears to be little evidence-based information. A shift from case reports to systematic evaluation and from descriptive to prescriptive research is needed. The discipline of cytological diagnostics requires greater academic interest, and better guidance and structuring. PMID- 11433658 TI - [Diarrhea due to Clostridium difficile toxin in hemato-oncological patients]. AB - In two patients with multiple myeloma, men aged 72 and 54 years, diarrhoea developed upon chemotherapy with vincristin, doxorubicin and dexamethasone (VAD). In the second patient, diarrhoea developed after subsequent peripheral stem cell mobilisation. Pseudomembranous colitis was seen in the first patient during endoscopy but an enzyme immunoassay of the faeces was false negative for Clostridium difficile enterotoxin. The bacterium was later cultured from stool samples and toxins were detected in a repeated immunoassay. Stool samples of the second patient were positive for C. difficile enterotoxin. For both patients an antibiotic treatment resulted in a rapid recovery. In haemato-oncological patients, diarrhoea is often caused by oncolytic therapy. However, consideration should also be given to C. difficile infection as an alternative cause which is easily treatable. PMID- 11433660 TI - [Gastrointestinal surgery and gastroenterology. XI. Stomas and stoma surgery]. AB - Stomas are an essential part of gastrointestinal surgery. Indications for stoma construction are faecal diversion from a distal diseased bowel segment, prevention of an intestinal anastomosis in intra-abdominal sepsis, and faecal incontinence. Pre- and postoperative counselling and nursing care is essential for a good functional outcome. Following stoma construction, complications such as dermatitis, retraction, prolapse, stenosis and parastomal hernia occur in 30 60% of cases. Thirty percent of stomas need surgical re-intervention in the first 10 years. For diversion of a distal anastomosis, construction of a loop-ileostomy is preferred to a loop-colostomy. Closure of a temporary stoma should not be done within eight weeks of construction. Preoperative evaluation of the distal segment is mandatory. Stoma closure involves an intra-abdominal anastomosis with all its associated complications. The incidence of complications after stoma closure is about 10%. PMID- 11433661 TI - [Gastrointestinal surgery and gastroenterology. XII. Care of stoma patient]. AB - A stoma nurse provides information, counselling and care to patients with gastrointestinal and urological conditions such as a stoma, incontinence, fistulas or an ileoanal anastomosis with a pouch. The stoma nurse will provide the patient and his or her partner with pre-operative information, post-operative counselling, practical care and instruction as well as check-ups at the outpatients' clinic. Careful selection of the most suitable collection device is highly important. This stoma care has a positive effect on the quality of life of a patient with a stoma. The most common complication is peristomal dermatitis which, in combination with care problems, may lead to social restrictions. Providing the patient's physical and psychological health is not in danger, the stoma care nurse can treat or control these complications, thereby avoiding the need for further surgery. PMID- 11433662 TI - [From gene to disease; mutation in mitochondrial DNA and maternally inherited diabetes mellitus with deafness (MIDD)]. AB - MIDD is a maternally inherited disorder with diabetes and impaired hearing due to a reduced perception of high tones. The disorder is caused by an A to G mutation at position 3243 in mitochondrial DNA. Approximately 1.3% of insulin-dependent diabetic patients in the Netherlands has this mutation. The main defect in these patients seems to be a reduced secretion of insulin by the pancreas in response to glucose stimulation. PMID- 11433663 TI - [Diagnostic image (42). Solitary bone cyst of the left calcaneus]. AB - A 9-year-old boy presented with a painful left heel. A plain radiograph showed a lytic lesion with a benign appearance, with a simple bone cyst or an intraosseous lipoma as the most likely differential diagnostic possibilities. MRI demonstrated a simple bone cyst. PMID- 11433664 TI - [Health problems of homeless people attending the outreach primary care surgeries in Amsterdam]. AB - OBJECTIVE: To describe the characteristics and health problems of homeless people. DESIGN: Retrospective descriptive cross-sectional study at two primary care surgeries for the homeless in shelters in Amsterdam supported by the Municipal Health Service. METHOD: During the period April 1997 to August 1999, the socio-demographics, reasons for encounter, medication, substance use and prevalence of HIV and lifetime tuberculosis were recorded in a standardised manner. Referrals to a shelter infirmary at the Salvation Army or a general hospital were investigated. RESULTS: The study population (n = 364) had an average age of 43 years (SD: 12) and 84% of the population were men. During the first consultation the following problems presented: skin problems in 26% of the population (traumatic injury, infected wounds, tramp's feet, scabies, lice, abscess, cellulites and erysipelas) and respiratory infections in 21% of the population (sinusitis, acute bronchitis and pneumonia). Medication was requested by 9% of the population of which 63% preferred tranquillisers. Medication prescribed by other physicians was stated by 46% of the population (21% tranquillisers, 14% methadone). Alcohol dependency was present in 25% (n = 92) and drug dependency in 29% of the population (n = 105). Intravenous drug use was prevalent in 14% of the population. Of the drug users, 49% received addiction care as opposed to 2% of the alcoholics. HIV prevalence was 5% and the lifetime prevalence of tuberculosis was also 5%. Shelter infirmary admittance was indicated in 15% of the population (n = 53; 91% men), with a combination of trauma 38%, skin disease 38%, pneumonia 21% and tramp's feet 17%. Five homeless men (1.4%), alcoholic street dwellers, presented with various life-threatening health problems in an advanced stage. CONCLUSION: Homeless people in Amsterdam have poor health and suffer from various problems. Outreach primary health care adjusted to the lifestyle and multiple health problems of the homeless was clearly very important for public health. Homeless alcoholics were not using addiction care services. PMID- 11433665 TI - [High prevalence of mental disorders and addiction problems among the homeless in Utrecht]. AB - OBJECTIVE: To determine the prevalence of psychological disorders and addiction problems amongst homeless people in Utrecht, the Netherlands. DESIGN: Descriptive cross-sectional study. METHOD: In 1998, the Trimbos Institute in Utrecht held 150 interviews with adult homeless people by means of a random sample taken from six centres for the homeless. The following screening instruments were used to determine the prevalence of a depressive episode, schizophrenia and other non affective psychotic disorders, an antisocial personality disorder and an alcohol or drugs dependency/abuse problem: the depression screener from Schrijvers et al., the schizophrenia section from the Composite International Diagnostic Interview (CIDI) a modified questionnaire from Schrijvers, related to the DSM-III R, about aggressive behaviour and the Addiction severity index, European variant, version III. By means of comparative research, optimum threshold values for the presence of the aforementioned disorders were determined with respect to extensive diagnostic interviews that were taken to be the gold standard. RESULTS: In the homeless population studied (n = 150 (response: 68%); 138 men and 12 women) 32% had a more narrowly defined form of depression, 15% a schizophrenic disorder (DSM-III-R-codes 295.00-295.70) and 52% an antisocial personality disorder. In the case of depression and schizophrenia it concerned a 6-month prevalence and in the case of antisocial personality disorders it concerned the lifetime prevalence. The lifetime prevalence of alcohol dependence or abuse was 59% in the population studied (n = 131) and the monthly prevalence was 22%. The lifetime prevalence of drug dependence or misuse was established to be 76% and the monthly prevalence 54%. The prevalences were high in comparison with international figures. A so-called double diagnosis was identified for 27% of the homeless population. CONCLUSION: Homeless people in Utrecht suffer from a poor mental health. PMID- 11433666 TI - [Tuberculosis of the pancreas in a HIV-seropositive patient]. AB - A 34-year-old man from Nigeria who had resided permanently in the Netherlands for five years had experienced fever, upper abdominal pain and weight loss for several months. He did not give the impression of being ill. A CT scan gave cause to suspect pancreatitis. An HIV test gave a positive result. Puncture of the accumulated fluid around the pancreas led to the diagnosis 'tuberculosis' (infection by Mycobacterium tuberculosis). Once the patient had made a good recovery with antituberculosis therapy, antiretroviral therapy was initiated, whereupon the number of CD4+ cells in the blood increased. Extrapulmonal tuberculosis is not unusual in HIV seropositive patients from countries with a high prevalence of tuberculosis. However, in such patients isolated tuberculosis of the pancreas is unusual and has not previously been described in the Netherlands. The diagnosis can be established following a CT guided puncture; tuberculosis is instantly suspected if the Ziehl-Neelsen stains are positive and the diagnosis can then be confirmed by a polymerase chain reaction (PCR) analysis and by culturing. Anti-retroviral therapy is withheld until response to anti tuberculosis treatment is satisfactory. PMID- 11433667 TI - [CBO guideline 'Bacterial meningitis']. PMID- 11433668 TI - [CBO guidelines 'Bacterial meningitis']. PMID- 11433669 TI - [Activated protein C, coagulation, inflammation and treatment of severe sepsis]. PMID- 11433670 TI - [Activated protein C, coagulation, inflammation and treatment of severe sepsis]. PMID- 11433671 TI - [Diagnostics in clinically occult, radiologically suspect breast lesions: more often surgical than needle diagnostics with image monitoring]. PMID- 11433672 TI - Functional interactions within the newborn brain investigated by adaptive coherence analysis of EEG. AB - To assess functional cortical organization in newborns during quiet sleep we investigated instantaneous interhemispheric coherences in six healthy full-term subjects using linked ear reference and average reference. Trace alternant was the most frequent EEG pattern during quiet sleep of these newborns and consists of burst and interburst periods. The calculation of coherence was performed by means of time-variant autoregressive models on the basis of Kalman filtering. Coherence changes simultaneously with the appearance of these burst and interburst periods. The highest level of coherence was observed during burst periods in comparison with interburst periods. The lowest level of coherence was observed just before the burst started. During burst, maximal coherence was reached at different moments--late in the low frequency band (0.5-1.5 Hz; about 3 s after the burst started) and earlier in higher frequency bands (> 2 Hz, about 2 s). Interhemispheric coherence depended on the region being investigated. A significant level of coherence could be observed over frontal, temporal and occipital regions. Our data demonstrate significant modification of interhemispheric coherence during quiet sleep of full-term newborns and, irrespective of the immaturity of the brain, these coherences differ significantly between cortical regions. PMID- 11433673 TI - [The recording of cognitive evoked potentials during and after transient global amnesia: report of three cases]. AB - It is of interest to record event-related potentials in the course of transient global amnesia (TGA) because the hippocampus and diencephalon, generally considered to be the sites of the dysfunction responsible for the amnesic episodes are also considered as two possible generators of the P300 wave. However, the only four cases reported so far in the literature showed an intact auditive P300 in three cases and an intact auditive P300 with reduction of visual P300 in one case. Here are reported four new cases. The P300 wave was readily identifiable in all four cases, without any amplitude reduction, thus suggesting that the condition did not entail inactivation or functional depression of P300 generators. Concerning P300 latency, in one case it was delayed but became normal after the ictus. In the second case, the latency, although within normal limits, shortened after the ictus. In the third and the fourth cases, the latency, initially within normal limits, remained unchanged. These apparently disparate results should be analysed in the light of the results of isotope measurement of cerebral blood flow during the amnesia, which are also inconsistent but most frequently indicate bilateral temporal or thalamic flow reduction. It remains to be determined in the future whether the stability or change in the P300 will make it possible to predict the brain region involved in transient global amnesia, which could perhaps vary from one patient to another. PMID- 11433674 TI - Auditory event-related potentials (P300) in epileptic patients. AB - Auditory event-related potentials (AERPs) were recorded during an auditory oddball paradigm in 108 epileptics and in 32 healthy controls. P300 latency varied in relationship with age only in controls. Symptomatic epileptics had significantly prolonged P300 mean latency compared to those without detectable brain lesion(s) on MR scan. Moreover, these latter patients were compared on the basis of epilepsy duration, type of seizure, seizure frequency and antiepileptic treatment; the application of a multiple regression model showed a significant relationship between P300 latency prolongation and epilepsy duration, seizure frequency and polytherapy. PMID- 11433675 TI - [Development of the central nervous system in mammals]. AB - In humans, the nervous system is induced during the third gestational week by molecular signals coming from the mesoderm, which modulate the temporal and spatial expression of specific genes in the cells of the dorsal ectoderm. The induced neural plate closes to form the neural tube where the cells actively proliferate in the germinal zone. The neuroblasts which have completed their last division migrate along the fibers of the radial glia to which they adhere, and this movement is essential to establish the normal cerebral organization. The regional identity of the developing brain is governed by the expression of homeobox genes, and the main central structures are clearly delineated by the end of the fifth week. The cerebral cortex begins to form on the seventh week, and the early specification of the cortical areas, which is under genetic control, would be modulated later on by environmental influences. When the neurons have reached their final position, they extend an axon, using surface molecules or diffusible molecules present along its pathway as cues to reach the appropriate target and form a synapse, and this process is a critical step for the establishment of neuronal relationships. The maturation and stabilization of neural networks is characterized by the apoptotic death of roughly 50% of the neurons, due to insufficient neurotrophic support, and by the remodeling of the initial synaptic connections in the surviving neurons. These regressive events occur late in development and depend on both the interactions with the environment and the resulting neuronal activity. PMID- 11433676 TI - Age-related changes of evoked potentials. AB - The aim of this review is to analyse the current state of our knowledge on evoked potentials (EPs) in ageing and to report some conclusions on the relation between EPs and elder age. Evoked potentials provide a measure of the function of sensory systems that change during the different stages of life. Each sensory system has its own time of maturation. The individuation of the exact period of life when brain ageing starts is difficult to define. Normally, the amplitude of EPs decreases, and their latency increases from adult to elder life. Many authors speculate that these modifications might depend on neuronal loss, changes in cell membrane, composition or senile plaques present in older patients, but there is no evidence that these changes might modify the cerebral function in healthy aged individuals. This review emphasises some incongruities present in different studies confirmed by daily neurophysiologic practice. Different techniques as event-related desynchronization (ERD), contingent negative variation (CNV) and Bereitschaftspotential, are available to study central neuronal changes in normal and pathologic ageing. PMID- 11433677 TI - Histological analysis and staining techniques modified and verified on ancient mummified tissues to study microorganism infestations. AB - The purpose of this work is to give a brief account of the possibility to estimate the preservation of human mummified tissues using histological analysis. This method can be useful to identify injuries and to plan qualified conservative actions on ancient human remains. Some preliminary results are presented here regarding the study on 16 ancient mummies from the Egyptian Museum of Turin. Samples of mummified tissues were taken without damaging the remains; they were hydrated again and dyed with histological techniques which were specifically modified and verified in same cases. Our research identifies some agents (biological and chemical) that appear to be responsible, among others, of mummified tissue destruction. The microscopic examination reveals features that might refer to fungal and bacterial infestation. Using special staining methods on seried sections we were able to suspect, at least in one case, the presence of biologically active forms. Microbiological assays confirm the vitality of fungi. Histological tissue analysis can then be useful to guide any conservative intervention for preservation and protection of the integrity of biological remains from museum collections. PMID- 11433678 TI - Temperature influence on stimulated PMN respiratory burst. AB - Body temperature can modulate the pathogenesis of infectious, metabolic and autoimmune diseases. This effect has been attributed to several hypothesized mechanisms. Body temperature could play an important role in influencing some cellular functions of human white blood cells. In this work we examined the temperature effect on the respiratory burst in human neutrophils. Human polymorphonuclear leucocytes (PMN) were obtained from heparinized venous blood by dextran sedimentation and erythrocyte lysis with NH4Cl (0.87%). Granulocytes were stimulated with opsonized zymosan (OZ), formyl-methionyl-leucyl-phenylalanine (FMLP), phorbol myristate acetate (PMA), and monosodium urate (MSU) crystals at different temperatures (26, 37, 39, 40, 42 degrees C). The technique of luminol dependent chemiluminescence (CL) was used as indicator of oxygen free radicals (OFR) release by stimulated cells. OFR production from PMN stimulated with OZ, PMA, FMLP was higher at 37 degrees C than at 26, 39, 40, 42 degrees C (p < 0.001 OZ stimulated PMN at 40-42 degrees C; p < 0.05 PMA stimulated PMN at 42 degrees C. Significantly different from 37 degrees C value). OFR release from PMN stimulated with MSU crystals was significantly increased at 39 degrees C compared to 37 degrees C value (p < 0.001). This effect could not only be attributed to temperature influence on neutrophil activity. The specific polymorphonuclear leukocyte response to the microcrystals and the temperature influence on chemical and physical characteristics of the crystals may play an important role. We are now studying the temperature effect on activity of PMN exposed to others crystals. PMID- 11433679 TI - Effect of hydrostatic pressure on morphological and ultrastructural aspects of normal and osteoarthritic human articular chondrocytes. AB - In this work we studied the morphological and ultrastructural aspects of normal and osteoarthritic (OA) human articular chondrocytes cultivated in alginate gel for 48 hours. After this period the chondrocytes in Petri dishes were exposed to cyclic pressurization (minimum pressure 1 MPa and maximum pressure 5 MPa) at 0.25 Hz frequency for three hours. In other loading procedures the cells were exposed to continuous pressure (24 MPa) for three hours. Some dishes were not pressurised and these served as controls. The cells were then fixed for transmission electron microscopy (T.E.M.) and for scanning electron microscopy (S.E.M.). No ultrastructural changes were observed in normal chondrocytes exposed at physiological pressure. OA cells placed under physiological pressure showed a partial recovery on morphological and ultrastructural aspects. Normal and OA samples exposed to continuous pressure (24 MPa) showed a morphological worsening in both T.E.M. and S.E.M. studies. PMID- 11433680 TI - Hippocampal synaptic plasticity: effects of neonatal stress in freely moving adult male rats. AB - The present study examines the effects of neonatal isolation on hippocampal LTP in adult male rats. Changes in dentate granule cell population measures, i.e., EPSP slope and population spike amplitude (PSA), evoked by tetanization of the medial perforant pathway were used to assess the effects of neonatal isolation on LTP over a period of 96 h. Following tetanization significant group differences were obtained for input/output (I/O) response measures of EPSP slope and PSA, with isolated males showing consistently higher values than in the other two groups. Comparisons made at 1 h post-tetanization (establishment of LTP) indicated that isolated males showed significantly greater enhancement than any other group. At 96 h (maintenance of LTP), however, neonatally isolated males showed significantly greater enhancement than either non-isolated siblings or unhandled controls. Additionally, isolation resulted in prolonging the duration of enhancement obtained from males. Thus, males show different enhancement profiles with respect to both the magnitude and duration of LTP and neonatal isolation alters these profiles in profound manner. PMID- 11433681 TI - Effects of clodronate on synovial fluid levels of some inflammatory mediators, after intra-articular administration to patients with synovitis secondary to knee osteoarthritis. AB - Recent studies have outlined the role of bisphosphonates, and particularly clodronate, as potential therapeutic agents for inflammatory and degenerative joint diseases. On this basis, we carried out an open, non comparative pilot trial to evaluate the effects of clodronate on synovial fluid concentration of some inflammatory mediators (prostaglandin E2, leukotriene B4 and tromboxane B2) after intra-articular, repeated administrations in 20 patients (7 males and 13 females) with synovitis secondary to knee osteoarthritis. At the end of the treatment period, statistically significant reductions (p < 0.05) of spontaneous pain and pain on active movement, evaluated by means of a 100 mm visual analogue scale (VAS), were reported. Linear regression analysis showed that the decrease of pain was correlated with the bisphosphonate induced reduction of prostaglandin E2 levels. These results, in spite of the limitation due to the open design of the trial suggest a possible role of bisphosphonates in the treatment of knee osteoarthritis. PMID- 11433682 TI - Termination of upper limb movement by cutaneous afferents. AB - We tested whether cutaneous afferents from the skin field close to an upper limb muscular region would carry information to spinal neurons at the onset or at the offset of a voluntary elbow extension movement lasting 1 s. We detected a depression of EMG activity both at onset and at the offset of the reaching movement but in the latter case depression was significantly larger and immediate. The marked depression of EMG activity suggests an inhibition, via spinal neurons, of the descending excitation to the motoneurones supplying the triceps brachii. This spinal control might be a very efficient mechanism for the termination of voluntary movement. PMID- 11433683 TI - Verbal communication in prosthetic dentistry. Input-process-outcome. PMID- 11433684 TI - [Congenital toxoplasmosis. Transitory negative serology]. AB - OBJECTIVES: Toxoplasmosis serology may become temporarily negative in children with congenital toxoplasmosis, leading to a risk of misdiagnosis and inadequate surveillance. The purpose of our work was to better understand the time course of toxoplasmosis serology which has not been studied specifically and to propose practical recommendations. PATIENTS AND METHODS: We conducted a prospective study in 217 children born with congenital toxoplasmosis between January 1988 and December 1997. Clinical, ophthalmological and serology data were collected every three months during their first year of life then every six months until three years of age and every year thereafter for all patients. Negative serology was defined as the absence of IgG at indirect immunofluorescence and ELISA (enzyme linked immunosorbent assay) and by the absence of IgM at ISAGA (immunosorbent agglutination assay). RESULTS: During the mean follow-up of 66 +/- 33 months (range 12-126 months), 33 children (15%) presented a period where the toxoplasmosis serology (ELISA and indirect immunofluorescence) was negative for a transient period reaching a mean 5 months. The dye test was performed in 25 of these children and was negative in 6 (24%). Among the negative conversions observed at routine testing, 73% occurred in children taking pyrimethamine/sulfadoxin therapy and the others occurred a mean 11.7 months after interruption of treatment. There was a positive association between maternal treatment and transient seronegativity in the cases where the maternal contamination had occurred during the first 2 trimesters of pregnancy. The serology became positive again in 30 of the 33 children (91%) and in 22 children there was a rebound. At last follow-up, the 3 other children still had negative serology (mean duration 35 months, range 3-62 months). CONCLUSION: Transient negative toxoplasmosis serology is a frequent phenomenon in children with congenital toxoplasmosis. Although the underlying pathophysiological mechanism remains unknown, it is crucial to avoid questioning the initial diagnosis of congenital toxoplasmosis and to continue regular routine monitoring. PMID- 11433685 TI - [Volvulus of the mobile spleen. Conservative laparoscopic treatment]. AB - BACKGROUND: Volvulus of the mobile spleen is a rare condition generally manifested by acute sudden onset abdominal pain localized in the left flank. Ultrasound or computed tomographic imaging generally provides the diagnosis. CASE REPORT: An 18-year-old patient was hospitalized for acute pain of the left flank that had begun 48 hours earlier. The abdominal scan visualized a volvulus of a mobile spleen. Laparoscopic distortion was successful, allowing normal recoloration of the spleen. DISCUSSION: Rapid intervention is crucial for conservative treatment. Laparoscopic treatment allows distortion manoeuvres and splenopexia using a mesh. This is the first reported case of successful conservative laparoscopic treatment of volvulus of a mobile spleen. PMID- 11433686 TI - [Ulceronecrotic purpura revealing Waldenstrom's macroglobulinemia]. PMID- 11433687 TI - [Genital infection with Oligella urethralis]. PMID- 11433688 TI - [Thalidomide and thrombosis: three observations]. PMID- 11433689 TI - [Acute rhabdomyolysis revealing corticoadrenaloma]. PMID- 11433690 TI - [Mycobacterium avium intracellular adenopathy in an immunocompetent adult]. PMID- 11433691 TI - [Carotid endarterectomy and prevention of cerebral ischemic events in the elderly]. PMID- 11433692 TI - [What is a Caucasian? Race and ethnicity in the medical literature]. PMID- 11433693 TI - [Aclotine (antithrombotic]. PMID- 11433694 TI - [Epidemiology of cervical papillomavirus infections. Recent knowledge]. AB - GENERAL DATA: There is now considerable evidence that high risk human papillomaviruses (HPV), such as HPV 16, are closely associated with cancer of the cervix. HPVs that are primarily transmitted through sexual contact, are found in over 99% of the cases of invasive cervical cancer. Although most women can be infected during their sexual life, a small minority is at risk for developing cancer. The long latency period between primary infection and cancer emergence suggests that additional factors are involved in the process of tumor development: sexual behavior, immune status, genetic predispositions, nutritional status, tobacco use, socio-economical level. NATURAL HISTORY: HPVs infect epithelial cells of the transformation zone of the cervix. As with other sexually transmitted diseases, the incidence of HPV infection is highest among young women. However, this viral infection is more often than not transient, because most individuals develop an effective type-specific immune response. Approximately 1% of the population has genital warts and 4% of women have cervical precancerous lesions: low grade squamous intraepithelial lesion (LGSIL) or high grade SIL. These last lesions preferentially observed in women aged 35-40 yrs are at high risk of progression towards an invasive cancer. ONCOGENIC POTENTIAL OF HPV: Pre-malignant and malignant cells arise as a result of HPV DNA integration in the host cellular genome, and overexpression of the viral E6 and E7 oncogenes. Cells acquire a proliferative advantage by escaping growth control exerted by p53 and p 105Rb. Both cellular proteins are indeed inactivated respectively by E6 and E7 proteins. Aneuploidy and karyotypic abnormalities are also key events in the tumor progression. A PREVENTABLE DISEASE: Cervical cancer is more than ever a preventable disease. While waiting for clinically applicable vaccination programs, strategies to prevent cervical cancer include 1) improved screening covering the widest possible population and using HPV testing, 2) close management and follow-up of women with precancerous lesions. PMID- 11433695 TI - [Acute alcoholic hepatitis: treatments]. AB - PROGNOSIS: Acute alcoholic hepatitis (AAH) is a severe form of alcohol-related liver disease with a high short-term mortality that can reach 50%. Long-term outcome depends on definitive weaning from alcohol and the development of cirrhosis. ESSENTIAL THERAPEUTIC STEP: Abstention from alcohol is the number one therapeutic measure required for treating AAH. Abstention must be total and definitive. THERAPEUTIC STRATEGIES: The pathogenic mechanisms involved in AAH have led to close assessment of numerous treatment protocols. Thirty-three randomized trials have evaluated drug treatments based on various strategies: antiinflammatory action using corticosteroids or colchicine; reduction of the hypermetabolism using propylthiouracil; hepatoprotective effect against oxidative stress using cyanidalol, alpha lipoid acid, silymarine, amlopidine, malotilate; vasodilatation to improve oxygenation of the centrolublular region using a calcium channel inhibitor, amlopidine; increased liver regeneration using anabolism steroids, intravenous perfusion combining insulin and glucagon; antifibrosis action using colchicine, D penicillamine; improved microcirculation due to increased deformability of the red cells and inhibition of TNF-alpha using pentoxifyllin. Eleven therapeutic trials have investigated the effect of parenteral or enteral artificial nutrition. GOLD STANDARD TREATMENT: Among all these strategies, the only one with a proven efficacy is corticosteroid therapy. Four trials have demonstrated the effect of corticosteroid therapy on short-term survival and 3 of the 4 meta-analyses devoted to the topic have demonstrated the usefulness of corticosteroid therapy in severe forms defined by a Maddrey index > or = 32: bilirubin in mumol per liter/17 + 4.6 (patient's PT in seconds--control PT in seconds) and the presence or not of encephalopathy. The gold standard treatment for severe AAH is oral prednisolone 40 mg/d for 1 month (excluding contraindications). PERSPECTIVES: Despite the effect of corticosteroid therapy, mortality at 2 months in severe AAH is still about 30%. Recent experimental data suggest that monoclonal anti-TNF alpha antibodies could be useful. PMID- 11433696 TI - [Generalized tonic-clonic status epilepticus: therapeutic strategy]. AB - BASIC STRATEGY: Generalized tonic-clonic status epilepticus is a medical emergency requiring very rapid administration of anti-epilepsy drugs to avoid or prevent neurological damage. First intention treatment is based on rapid-action intravenous benzodiazepine (BZD) associated with another long-action anti epilepsy drug. General anesthesia with respiratory assistance may be needed if the seizures are refractory. We considered the pharmacodynamic, pharmacokinetic and pharmacoeconomic properties of drugs proposed for the treatment of status epilepticus. TREATMENT EFFICACY: An analysis of the literature and clinical practice show that, used alone, BZDs have a rapid effect and are effective in 54 to 84% of the cases. When hydantoins are combined with BZD, cessation of seizures can be achieved in 94% of the patients compared with 82% when phenobarbital is used alone. However, the administration of hydantoins requires 15 to 30 min whereas phenobarbital is effective in 5 minutes. Irrespective of the type of BZD combined with hydantoins, no difference has been observed concerning clinical efficacy. Midazolam appears to be as effective as barbiturics. Cardiac function must be monitored when hydantoins are used although admission in an intensive care unit may not be required, unlike the situation with phenobarbital that may lead to intubation. IN CLINICAL PRACTICE: Considering non-refractory status epilepticus, a comparison of the efficacy of the proposed drugs, their side effects and their cost demonstrates a good cost/benefit ratio for phenobarbital and good tolerance for fosphenytoin. If cessation of the seizures cannot be achieved, other therapeutic strategies may have be to used: induction of barbituric coma with thiopental, general anesthesia using propofol, or midazolam or lidocaine. PMID- 11433697 TI - [Lymph node recurrence in breast cancer. Report of 1,119 cases of infiltrating cancer]. AB - PURPOSE: To analyze the main determinants of lymph nodal recurrence in early breast cancer and to compare our results with literature data. METHODS: Retrospective analysis of 1119 patients with stage I-II infiltrating breast cancer treated by conservative radiosurgical association from 1980 to 1992, with a 10-year median follow-up. RESULTS: The global rate of lymph nodal recurrence (LNR) was 2.7% (30 cases). Age under 40, high histoprognostic grade and previous local recurrence were the main risk factors of LNR. Axillary nodal involvement at time of diagnosis, number of lymph nodes sampled and tumor localization were not significantly related to LNR. CONCLUSION: Lymph nodal recurrences of early breast cancers are not frequent, but their prognosis is very unfavourable. A multidisciplinary approach of these LNR is necessary, taking into account previous treatments. PMID- 11433698 TI - [Glutamate antagonists in neurology]. AB - Glutamate is the major excitatory neurotransmitter of the central nervous system. Besides its importance in many physiological processes, increased glutamate release and subsequent excessive stimulation of the various glutamate receptors are thought to play critical roles in the pathophysiological mechanisms underlying many neurologic diseases. Experimental data suggest that blockade of glutamate receptors or inhibition of glutamate release has positive effects in many disease models. Glutamate antagonists are already in clinical use for the treatment of Parkinson's disease, epilepsy, spasticity, and neuropathic pain. Overall, glutamate antagonists have not been found clinically effective for neuroprotective treatment of cerebral ischemia or chronic neurodegenerative diseases, with one exception. Side effects of glutamate antagonists can be mainly attributed to central mechanisms and include psychosis, agitation, and disorientation. It is to be hoped that further development of new glutamate antagonists that block disease-relevant subtypes of glutamate receptors will lead to more effective drugs with fewer side effects. PMID- 11433699 TI - [Spontaneous occlusion of the circle of Willis (moyamoya disease). Diagnosis and therapy]. AB - Spontaneous occlusion of the circle of Willis (moyamoya disease), a rare cerebrovascular disease, may lead to repeated hemodynamic stroke due to unremitting changes of the basal cranial vessels. Moyamoya can be diagnosed on its specific epidemiological, clinical, and radiological features. In patients with distinctive Moyamoya disease, improvement of cerebral blood supply by means of neurosurgical revascularization enables definitive treatment. This review summarizes current knowledge of the clinical and diagnostic features of this disease and illustrates its pathological and pathophysiological characteristics. Additionally, indications and techniques of current neurosurgical revascularization procedures are illustrated. PMID- 11433700 TI - [Pathophysiology, clinical aspects and therapy of sleep disorders in Parkinson disease]. AB - Parkinson's disease (PD) is associated with sleep disorders which are attributed mainly to dopamine deficiency, nocturnal akinesia, drug therapy, and cofactors such as age and depression. These disturbances affect the macro- and microstructure of both REM and non-REM sleep and motor, respiratory, and autonomic functions. Excessive daytime sleepiness and the interactions between sleep and daytime motor performance in PD are not yet completely understood. Correct diagnosis and treatment of sleep disorders is essential due to the risk of harm to the patient and others and due to their effect on quality of life for all concerned. As sleep disorders in PD are extremely common (about 70%) and may have severe consequences, a systematic sleep history and specific therapy should be considered integral to treatment in every PD patient. PMID- 11433701 TI - [Restless legs syndrome and periodic limb movements in sleep]. AB - Restless legs syndrome is one of the most common neurological disorders, with a prevalence of 2% to 9% in the elderly population. Sensory and motor symptoms of the legs and an urge to move that occur at rest may lead to severe sleep disturbances and are part of the syndrome. Typical history and normal neurological examination lead to the clinical diagnosis. Additional laboratory and neurophysiological investigations are necessary to rule out associated diseases. The indication for polysomnography to record periodic limb movements in sleep must be discussed in individual cases. Treatment strategies will be recommended individually according to the disease severity. In this article we present an overview of the clinical symptomatology and include recommendations on diagnosis and treatment of RLS and differentiation of RLS from periodic limb movement disorder. To this purpose, the Motor System and Sleep Work Group of the German Society of Sleep Medicine presents modified guidelines for diagnosis and treatment of RLS according to recent recommendations of the American Sleep Disorder Association. PMID- 11433702 TI - [Disorders in gaze saccades in Huntington disease. Clinical correlations]. AB - Oculomotor abnormalities have long been recognized in Huntington's disease (HD). The precise correlation between them and other clinical findings has not yet been determined. Using videonystagmography, we studied reflexive, visually guided horizontal saccades in 32 patients with genetically confirmed HD: nine female and 23 male patients, including six with young onset HD (YOHD), 19 with adult onset HD (AOD), and seven with late onset HD (LOHD). Huntington's patients exhibited increased saccade latency (P < 0.05), decreased saccade velocity (P < 0.0005), and impaired saccade accuracy (P < 0.01). A significant difference between the different groups of patients could be determined, and YOHD was characterized by normal latency and decreased saccade velocity while LOHD showed increased saccade latency but normal velocity. Furthermore, we found a significant difference between the genetic data (length of CAG-repeats) and saccadic abnormalities, with higher repeat numbers corresponding to shorter latency and decreased velocity, as in YOHD. The study of saccade parameters might be useful as an objective method for testing the effectiveness of future therapies. PMID- 11433703 TI - [Spontaneous herniation of the thoracic spinal cord as the etiology of progressive Brown-Sequard syndrome. A description of 3 cases]. AB - Spontaneous herniation of the spinal cord through a ventral dural defect is a rare neurological entity. We report on 3 patients treated in our clinic within 6 months. In 2 cases, the lesions were located at the BWK6 level and, in one case, at BWK2. The clinical presentation of all patients was f progressive Brown Sequard's Syndrome. Diagnosis was confirmed by magnetic resonance imaging. In all cases, operation was performed. The myelon was replaced in the dural sleeve and the defect covered with a patch of lyophilized dura. After 1 year, symptoms were stable in 2 patients. In one case there was a slight deterioration because of an inexplicable swelling of the spinal cord which could not be controlled with a second operation. Pathophysiological, clinical, and radiological aspects are presented and discussed. PMID- 11433704 TI - [Intravenous immunoglobulin and prednisolone treatment of cryoglobulinemic polyneuropathy]. AB - A 71-year-old man with cryoglobulinemia associated with severe symmetrical sensorimotor polyneuropathy was successfully treated with intravenous immunoglobulin (IVIG) and a corticosteroid. Within 2-3 months, he developed distal motor and sensory deficits and burning feet deteriorating after cold exposure. On examination, symmetric hypesthesia and hypalgesia were found along with bilateral loss of vibration and position sense, loss of ankle jerks, and paralysis of the pretibial muscles with step-page gait. The laboratory exam revealed elevated cryoglobulin levels and reduced complement C4. Nerve conduction studies were consistent with a severe axonal sensorimotor polyneuropathy. Sural nerve biopsy showed chronic neuropathy with an acute component. The patient underwent a combined treatment with prednisolone (initially 100 mg) and intravenous immunoglobulin. His condition markedly improved after almost a year of treatment. Unfortunately, the treatment with intravenous immunoglobulin had to be interrupted due to an allergic reaction after the sixth application. PMID- 11433705 TI - [Cerebellar syndrome, exophthalmos and secondary hypogonadism in Erdheim-Chester disease]. AB - We present a 50-year-old patient with a slowly progressive cerebellar syndrome, left-sided exophthalmos, secondary hypogonadism, and multiple pleomorphous skin alterations. The diagnosis of Erdheim-Chester disease was established by the radiological detection of a left-sided retrobulbar space-occupying mass, a hypophysial stalk lesion, alterations in both cerebellar hemispheres, retroperitoneal imbibition, osteolytic/osteosclerotic changes in the metaphysis and diaphysis of the long bones, and a skin biopsy with histological detection of a non-Langerhans-cell histiocytosis. The etiology of the Erdheim-Chester disease is unknown. Cerebral manifestations of this rare disease have been documented in only a very few cases. Whereas the extracranial alterations are due to pathologic histiocyte proliferation, cerebellar changes are considered to be the result of demyelinisation or infiltration of xanthogranulomas. PMID- 11433706 TI - [Reversible phenytoin-induced extrapontine myelinolysis]. AB - Antiepileptic drug (AED)-induced side effects still may hinder antiepileptic therapy. We describe a 38-year-old woman with complex focal seizures. Due to normal neuroradiological findings and the lack of evident pathogenetic factors, we diagnosed cryptogenic epilepsy. In response to cognitive side effects, the initial therapy with phenobarbital was replaced by phenytoin treatment only. Subsequent MRI investigation showed a hyperintense lesion in the corpus callosum without gadolinium enhancement. Responding to an increase in seizure frequency (SF), phenytoin was replaced first by lamotrigin monotherapy and then by additional valproate treatment. An MRI follow-up investigation did not show the previously described lesion of the corpus callosum, thus suggesting phenytoin induced extrapontine myelinolysis. In the literature, vigabatrin-induced demyelinization of the corpus callosum has also been described. In patients treated with the AEDs phenytoin or vigabatrin who demonstrate increased SF, such a side effect should be taken into account and MRI investigation performed. PMID- 11433707 TI - [Nerve growth factor (NGF) in treatment of diabetic polyneuropathy. One hope less?]. AB - Diabetic polyneuropathy (DPN) is by far the most common neuropathy in the western world. In Germany, some 3-4 million patients are afflicted. In the U.S., a large scale multicenter trial was initiated to evaluate the efficacy and safety of recombinant human nerve growth factor (rhNGF) in the treatment of DPN. A total of 1019 patients were randomly assigned in this double-blind study to receive either rhNGF or placebo over a 48-week period. The rhNGF had no significant side effects apart from mild hyperalgesia at the injection site. However, the substance failed to show significant benefit on global assessment, neuropathic sypmtoms and signs or neurophysiological parameters of nerve function when compared to the placebo group. Possible reasons for the negative outcome of this trial are the small dosis of rhNGF used, choice of a study population different from those enrolled in earlier promising studies, and the multifactorial etiology of DPN, which poses significant difficulties to the design and evaluation of therapeutic trials. PMID- 11433708 TI - [Recommendations for prevention of malaria in patients with epilepsy]. AB - Recommendations for malaria prophylaxis must balance the risk of malaria against the risk of side effects of malaria prophylaxis. In patients with epilepsy, the risk of inducing epileptic fits must also be considered. Mefloquine has been shown to have proconvulsive effects and induce seizures. For other drugs like atovaquone-proguanil, the experience up to now is insufficient in epileptic patients. Considering this balance of risk vs benefit, recommendations are given for malaria prophylaxis in patients with epilepsy. PMID- 11433709 TI - [Dostoievsky's life in the interrelation between epilepsy and literature]. AB - Fyodor Mihailovich Dostoievsky is considered one of the most important Russian authors of the nineteenth century. He suffered from epilepsy since childhood, which disease often was a theme in his novels. This paper describes Dostoievsky's life and examines the various epileptic protagonists appearing in his works. Finally, the author's own epilepsy and particularly its etiology are discussed. PMID- 11433710 TI - [Tau protein disorders--a new category of degenerative brain diseases]. PMID- 11433711 TI - [Position of the German Society of Neurology on limited approval of rt-PA for treatment of acute ischemic stroke]. PMID- 11433712 TI - [G-DRG: the for Germany modified AR-DRG system as a complete case-associated hospital reimbursement system according to section 17bKHG. Must it apply to psychiatry?]. AB - The introduction of a German ("German") DRG system (G-DRG) by 01.01.2003 is to improve efficiency and transparency of hospital performance and to terminate the presently implausible variability of costs. For the first time world wide the attempt is undertaken to finance by a DRG-system--supplemented by certain additional charges and rebates--hospital costs completely. Institutions submitted to the psychiatry personnel regulation (PsychPV) are excluded. As the basis for the G-DRG-system self-administration authorities have selected the Australian AR DRG system. The adjustment to German conditions is an extraordinary challenge: Compatibility must be achieved between the German classification of diagnoses (CGD-10) and procedures (OPS-301) and pertinent coding standards and the Australian classification systems. In the hospitals a unit cost accounting must be established, which at least approximately allows a strictly case-related calculation of actual costs. The relative cost weights of the DRGS and thus in the long run, their prices will be calculated on the basis the costs of a complete sample of cases of a representative subset of hospitals. The full-scale DRG system will confront with new risks. One is the transfer of treatment components and thus costs to Institutions not covered by the DRG system (e.g. rehabilitation hospitals, psychiatry). Thus, psychiatry will be at least indirectly involved. In addition, psychiatric patients will be directly affected if they are treated--possibly due to misallocation--in non-psychiatric institutions (e.g. internal medicine, neurology) or in psychosomatic departments not covered by the PsychPV. PMID- 11433713 TI - [Klaus Joachim Zulch (1910-1988). Comments on the issue cover picture]. PMID- 11433714 TI - [The hospital discharge card of diabetic patients]. AB - The diagnostic criteria and the classification of diabetes mellitus have been subject in the last few years to a new substantial revision. Moreover several clinical trials have demonstrated a clear correlation between glycaemic control and the onset and progression of micro and macrovascular complications. The glycaemic aims of therapy have therefore been defined. Diabetes should be subjected to a periodic screening for the early recognition of complications. These elements should orient the compilation of the clinical folder and the hospital discharge card of the diabetic patient. PMID- 11433715 TI - [Conventional colonoscopy and colonography with magnetic resonance in the diagnosis of endoluminal lesions of the colon]. AB - The diagnostic ability of Magnetic Resonance Colonography (MRC) in detecting colonic endoluminal masses was compared with that of conventional colonoscopy (CC) and related to the findings from histologic examination. In seventy consecutive patients MRC achieved a diagnostic accuracy similar to CC. Therefore MRC could be useful in screening patients at high risk for colonic cancer. However every patient with MRC-detected endoluminal lesion must undergo CC for histologic diagnosis. PMID- 11433716 TI - [Treatment of the hypertensive patient. A new organization model]. AB - A recent pharmaco-epydemiologic study has demonstrated the pitfall of the organization for the treatment of arterial hypertension in Italy. A possible solution of such a problem has been tested in Campania by organizing a telematic network in the Outpatient Center for Hypertension of the Department of Clinical Medicine, University Federico II in Naples, the Outpatient Unit for Hypertension of 25 Hospitals and 250 General Practitioners. Such an organization has allowed a significant reduction of the hypertensive patients admitted in Emergency or hospitalized; and an increase of the percentage of patients with blood pression below 140/90 mmHg. PMID- 11433717 TI - [Cyclosporin in atopic dermatitis]. AB - Cyclosporin A seems to became more and more important in the treatment of atopic dermatitis. Open, prospective study: 8 weeks of treatment and 6 months of follow up in wash-out. 15 patients were selected (10 males and 5 females) with mean age of 35.5. The patients were suffering of atopic dermatitis non responder to preceding treatments, and free from any pathological conditions contra-indicating the use of cyclosporin A. Cyclosporin A was orally administered at the dosage of 5 mg/kg/day for 8 weeks. The patients used a diary with a score from 0 to 3 for the following symptoms: extensions of skin lesions, itch and sleeping sickness. Treatment with cyclosporin A induced a significant improvement of the parameters evaluated. No significant side effects were observed. No relapses were recorded during the six months of follow-up. PMID- 11433718 TI - [Late onset immunodeficiency with hypo-IgG and hyper-IgM, T CD4+ lymphocytopenia and vitiligo]. AB - The Authors report the clinical case of a patient with a deficit of humoral immunity who developed infections since puberty. The serum levels of IgG and IgA decreased progressively in the fourth decade of life, while serum IgM increased. Moreover, the patient developed a marked CD4+ T lymphocytopenia and a meager B lymphocytopenia, vitiligo, positivity for anti-SSA/Ro autoantibodies and granulomatous phlogosis of the knee. The heterogeneity of the clinical and laboratory data suggests that this patient might present an overlap immunodeficiency syndrome with some of the clinical and immunological features typical of the hyper-IgM syndrome (in the X-linked or autosomal forms) and others that can be referred to a nosologically distinct humoral immunodeficiency such as the common variable immunodeficiency. PMID- 11433719 TI - [Chemotherapy in gastroenterologic neuroendocrine tumors]. AB - Gastrointestinal neuroendocrine tumors (carcinoids tumors and endocrine islet cell tumors) are a family of rare malignancies with many typical characteristics by a biologic, epidemiologic and clinical point of view. In this category of neoplasia an integrated clinical and therapeutic approach is mandatory, whereas for too many years these tumors were investigated and treated in an empirical way without considering an integrated approach. The singular features of this class of malignancies, with different and complex symptomatology, with enigmatic clinical presentation and outcome and with conflicting therapeutic options, oblige the clinicians to give different treatment to the patients. Chemotherapy has a marginal role in gastrointestinal neuroendocrine tumors mostly because it is used in patients with advanced disease not suitable for other therapeutic approach (surgery, thermoablation, chemoembolization, biotherapy). Unfortunately in the past it was not possible to establish the efficacy of chemotherapy in these malignancies because most of the studies pooled without distinction carcinoids, pancreatic tumors and hepatic metastases from unknown primary. The most extensively studied drugs have been streptozotocin, doxorubicin, mitoxantrone, dacarbazine, used alone or in combination; the gold standard today is considered the association of streptozotocin with doxorubicin or 5 fluorouracil, but there is strong evidence that the disappointing results in the treatment of these rare malignancies could be improved in a multidisciplinary fashion; in this field the combination of new drugs with aggressive surgery, radionuclide therapy, biotherapy and local therapeutic approach will give new opportunities to better control the symptoms and the clinical course of gastrointestinal neuroendocrine tumors. PMID- 11433720 TI - [Early diagnosis of pancreatic carcinoma]. AB - The poor prognosis of pancreatic carcinoma seems mainly due to the late symptoms onset. Age, cigarette smoking, family history of pancreatic carcinoma, chronic pancreatitis, a history of previous malignancy, new onset of diabetes mellitus without family history or overweight, and secondary failure to oral antidiabetic agents in a long standing type 2 diabetes mellitus are the hitherto identified risk factors. In consideration of the increasing availability of endoscopic ultrasonography--the most accurate examination in the diagnosis of small pancreatic masses--a perspective multicentric screening study on subjects with some of these risk factors may be justified at this time. PMID- 11433721 TI - [Guideline for the hemodialytic treatment of chronic uremia]. AB - In this review we have summarized the guidelines on hemodialysis prescription and dose in order to attempt of simplify an issue often difficult to understand for the majority of physicians. In brief, we start from the results of National Cooperative Dialysis Study and then we describe the Urea Kinetic Model, the KT/V and the Protein Catabolic Rate (PCR). Simplified formulas to obtain KT/V and PCR are also reported. At the end, the modalities related to postdialytic blood samples and connected problems are described. PMID- 11433722 TI - [Fatigue syndrome caused by malignant tumor. An increasing priority in patient care]. AB - As oncologists have become more effective in alleviating pain, nausea and depression, fatigue has emerged as the most important symptom suffered by cancer patients. Indeed, the current literature suggests that fatigue is currently the most important untreated symptom in cancer medicine. In recent surveys of patients and their caregivers, fatigue is more important for the quality of life than pain, nausea or depression. Yet these same surveys confirm that oncologists underestimate the importance of cancer related fatigue. This may be partly because patients often do not fully share the full nature of their concerns. When patients do raise the issue of fatigue, the physicians' recommendations are often non specific. However, recent research has shown that fatigue is not inevitable and untreatable, but a symptom amenable to differential diagnosis and specific intervention. Like pain, fatigue is intrinsically a subjective problem where the doctor relies on the patient's reporting. Weakness, exhaustion, lethargy and asthenia are all used as functional descriptions of fatigue. While these are descriptive terms, clinical research in the measurement and alleviation of fatigue requires reproducible measurement tools. Several studies already exist and have begun to explore this important area of symptom management. PMID- 11433723 TI - [Infectious diseases in the short stories written by Verga and Pirandello]. AB - The author invites to appreciate some short stories by Giovanni Verga and Luigi Pirandello, Italian writers who lived at the turn of the last century. These tales give an interesting description of two infectious diseases (malaria and cholera), and analyze their relationship with the conditions of poverty and ignorance spread across rural areas in Sicily--a region in the South of Italy--at that times. PMID- 11433724 TI - [The transparency commission: evaluation and re-evaluation]. AB - PURPOSE: The purpose of the French Transparency Commission is to provide scientific advice concerning the usefulness, interest and good use of drugs. DRUG APPROVAL: The work of the Transparency Commission lies at the interface between the French or European marketing approval procedures and the health and economic prerequisites of the French community. ASSESSMENT: The opinion of the French Transparency Commission is used to assess the medical service provided by a new drug and the improvement of this medical service subsequent to its use. This opinion is taken into consideration for establishing the reimbursement rate applied by the social security organizations and the selling price set by the administration. The expert opinions and recommendations established by the Transparency Commission participate in implementing good use of drugs. PMID- 11433725 TI - [Strong opioids for chronic non-cancer pain]. AB - A CHALLENGING SITUATION: A number of patients experiencing chronic noncancer pain are unsatisfied with standard treatment modalities. This raises the question of whether there may be a place for strong opioids in the management of these patients. Randomised placebo-controlled trials of strong opioids generated rather disappointing results in this type of pain. Observational studies have indicated that strong opioids may improve comfort and function in some patients with intractable nociceptive or neuropathic pain. However, opioids may be ineffective in others and intolerable side effects, heightened pain and functional impairment as well as drug addiction may also occur. A PROMISING SOLUTION: Finally, strong opioids do not appear to be the issue to all intractable chronic nonmalignant pain states, but they may be a possible issue to a subset of selected and informed patients who agree on the goals of the treatment and accept regular monitoring. PMID- 11433726 TI - [Phase I clinical trials]. AB - OBJECTIVE: Phase I clinical trials consist in studying tolerance to new drugs administered for the first time to humans, the pharmacodynamics of these drugs in order to describing as clearly as possible their pharmacological mechanism of action, and their pharmacokinetics in order to determine the metabolic and excretory pathways involved in the human organism. IMPLEMENTATION: Phase I trials are conducted in healthy volunteer men and women. An exception is made for drugs with predictable toxic effects, for example anti-cancer drugs, which cannot be tested in healthy subjects. These trials are controlled by legal regulations clearly defined in the French law of December 20, 1988. CONTRIBUTION: Data collected from Phase I trails in humans are crucial for initiating phase II and III drug development trials to be conducted in patients and designed to obtain a satisfactory assessment of the new drug's risk/benefit ratio. PMID- 11433727 TI - [Hyperlipidemia in the elderly]. AB - THE ELDERLY SUBJECT: Cardiovascular prevention trials have clearly established the beneficial effect of lipid-lowering drugs, basically statins, for adults, and in particular for adult males. There has not however been any study specifically analyzing prevention in the elderly. In addition, the relationships between high cholesterol level and cardiovascular risk in the elderly subject are complex and require specific adaptation with markers of poor health status. PREVENTIVE MEASURES: Even though the relationship between elevated total cholesterol or LDL cholesterol and cardiovascular risk is difficult to demonstrate in the elderly, analyses of subpopulations are beginning to show evidence that a lipid-lowering treatment can have a beneficial preventive effect in the elderly. Other research avenues also point to perspectives for stroke prevention or reduced risk of dementia in patients treated with statins. RECOMMENDATIONS: Until the results of ongoing clinical trials become available, recommendations for therapeutic strategies in the elderly are basically founded on clinical experience in light of current evidence. It is reasonable to recommend a lipid-lowering regimen for secondary prevention and to continue treatment beyond 70 years for primary prevention if the patient's overall cardiovascular risk warrants prevention. But, on the basis of current evidence, there is no rationale for recommending a large screening of hypercholesterolemia in primary prevention in subjects over 70 years of age. Likewise, taking into account associated disease states and the increased risk of drug interactions, primary prevention regimens cannot be recommended except for elderly patients with a high overall cardiovascular risk. PMID- 11433728 TI - [Hyperparathyroidism diagnosed during pregnancy]. AB - BACKGROUND: Diagnosis of hyperparathyroidism during pregnancy is rare because symptoms are often moderate and similar to signs observed during normal pregnancy. The gravity of such an association is probably underscored. CASE REPORT: The diagnosis of hyperparathyroidism was made at 9 weeks gestation in a young woman who declined surgery. She was seen again at 32 weeks gestation for in utero fetal death. Nine months later she accepted surgery and a parathyroid adenoma was removed. DISCUSSION: Hyperparathyroidism during pregnancy is associated with significant morbidity with major maternal or fetal complications in 50% of the cases. Surgery during the second trimester of pregnancy is the treatment of choice. If hyperparathyroidism is diagnosed in young women who desire pregnancy, surgery should be performed before advising pregnancy. PMID- 11433729 TI - [Ischemic neuropathy following surgery for an aortic dissection]. AB - BACKGROUND: Ischemic neuropathy is an uncommon and poorly-understood condition affecting the peripheral nervous system. Most cases involve the upper limbs and occur after vascular surgery. CASE REPORT: A 42-year-old man developed a multiple mononeuropathy of the lower right limb. The painful, axonal and predominantly sensory neuropathy was caused by ischemia and occurred subsequent to surgery for dissection of the aorta extended to the aortoiliac bifurcation. There was no sign of muscle or skin ischemia. The clinical course was slow and characterised by an incomplete recovery. DISCUSSION: Ischemic neuropathy of the lower limbs is generally observed after aortoiliac surgery. Occurrence after aortic dissection is exceptional. Most cases show a distal localization and are painful with little motor deficit. They must not be confused with global ischemia of the lower limb or radiculopathy. Early revascularisation is crucial for complete recovery. PMID- 11433730 TI - [Early migration of an implantable chamber catheter]. PMID- 11433731 TI - [Lymphocytic colitis in a women with Biermer's disease treated with Cirkan]. PMID- 11433732 TI - [Primary cardiovascular prevention: aspirin, vitamin E or placebo]. PMID- 11433733 TI - [An ethical decision-making tool. Training for decision-making in crisis or end of-life situations]. AB - We developed a patient-centered decision making tool to help healthcare teams make ethical decisions in crisis or end-of-life situations. This tool is the fruit of 15 years of healthcare experience and discussions on ethical issues with patients suffering from cancer, severe handicaps or terminal disease. It has been enriched by experience acquired since the publication of earlier work in the nineties. A three-step decision-making process is proposed providing a methodic aid for management decisions which remain unique for each individual patient. PMID- 11433734 TI - [Antivitamin K and paracetamol, an undesirable synergy]. PMID- 11433735 TI - [Anticoagulants in the elderly. Age-related factors]. AB - HEPARINS: For unfractionated heparins, certain studies have demonstrated increased hemorrhage rates correlated with age and with renal function. Subcutaneous injections every 12 hours are as effective and less uncomfortable for the patient than continuous intravenous infusions. Low-molecular-weight heparins are currently contraindicated for curative treatment and their use is not recommended for prevention when the creatinine clearance is below 30 ml/min. OTHER ANTICOAGULANTS: The dosage of sodium daparanoid should be adapted to the renal function with regular surveillance of anti Xa activity. With hirudin, there is a risk of product accumulation in case of renal failure. For oral anticoagulants the relationship between old age and the frequency of hemorrhagic complications is still a subject of debate. ORAL ANTICOAGULANTS: The starting dosage should be reduced to one-half or one-quarter of the usual dose. The INR should be measured regularly, particularly since elderly subjects are highly susceptible to wide variations in INR irrespective of the cause. It is also advisable to carefully determine appropriate situations when oral coagulants should be used. PMID- 11433736 TI - [Anticoagulants in the elderly. Indications]. AB - RISK AND BENEFIT: The beneficial effect of anticoagulant treatment is well established by numerous therapeutic trials demonstrating its usefulness in two main indications (venous thromboembolism and complete arrhythmia due to atrial fibrillation) in selected populations. Very few trials have been specifically conducted in elderly subjects and the increased risk of thrombosis and anticoagulant related bleeding cannot be used to extrapolate the results to elderly subjects. For this reason, a careful assessment of the initial risk and expected benefit must be made for each individual patient using a validated and systematic global evaluation system adapted for elderly subjects in order to better discern the indications and the risk of hemorrhage. PERSPECTIVES: The development of anticoagulation clinics, based on the method initiated by Professor Boneu at Toulouse, together with clinical trials designed to produce results transposable to the diverse and complex geriatric population, and finally the development of new oral compounds not requiring laboratory tests under the same conditions should help reduce the therapeutic risk for an equivalent benefit. PMID- 11433737 TI - [Aspects of background and discussion of qualitative research on the validity as scientific methodology]. PMID- 11433738 TI - [Drinking habits and characteristics of urban male residents with high alcohol intake--analysis of temporary housing residents after the Great Hanshin Earthquake]. AB - In December 1996, a health survey was conducted of 49,033 temporary housing residents of Kobe City, who were victims of the Great Hanshin Earthquake on Tuesday, January 17, 1995. A total of 33,414 residents (68.1% of those requested) filled in self-administered questionnaires. This study focused on males over 20 years old and examined factors related to increase in alcohol intake. A total of 14,179 men were analyzed, 67.4% of whom reported habitual drinking, and 32.1% an increase in their alcohol intake. The factors related to elevated alcohol intake were a low Breslow's health practice index, a high nervous complaint score and a low living score. In addition, the health practice index demonstrated an inverse link. There are many drinkers among urban residents and drinking related problems are easily evoked when living conditions worsen, as in the case of a natural disaster. Logistic regression analyses were used to predict which persons are likely to increase their alcohol intake. Significant predictors identified were the smoking habit, sleep disturbance, nervous complaints and the health practice index. It is important to ensure that this latter remains high to prevent the alcohol-related problems in urban residents. PMID- 11433739 TI - [Factors related to satisfaction level in daily life for older people]. AB - The purpose of this study was to examine relations between satisfaction levels and life-style in daily life for older people. A questionnaire, based on 7 factors of life satisfaction level and 13 factors of life-style chosen after considering theoretical validity, was administered to 1,320 healthy people aged 60 or more in the community (665 males and 655 females). Remarkable gender and grade differences were confirmed in the "physical health" satisfaction level. Satisfaction level for "personal relations" related to the number of friends for both sexes and to volunteer activities for males. The influence of the life-style factor on satisfaction level was highest in physical health. The influence of the number of friends was high for each satisfaction level. It was inferred that there are many aspects of life-style backgrounds contributing to the satisfaction level of older people in the community, and individual satisfaction with daily life is affected by different life-style factors. PMID- 11433740 TI - [Criteria for degrees of a smoothing method applied to mortality rates]. AB - PURPOSE: Age-period-specific mortality rates are smoothed by methods to clarify their characteristics and trends. In the present study a procedure to determine appropriate iteration times for weighted averages was devised, and examined by application to mortality data. MATERIALS AND METHODS: Mortality data for leukemia, solid neoplasms, and traffic accidents caused by cars in Japan were employed. The smoothing method was iterating weighted averages over age-period specific death rates and their four nearest neighbors in age-period data tables. The primary condition, which the iteration times should satisfy, was that the differences between crude data and smoothed ones were not significantly large, i.e., smoothed mortality data should be within 95% confidence intervals of crude rates. Therefore, the following steps were adopted to determine appropriate times: (1) estimation of 95% confidence intervals for crude age-period-specific mortality rates. (2) determination of iteration times under the primary condition that the proportion of cells left out of the confidence intervals should be around 5%. In cases where the proportion of cells outside of confidence intervals was still below 5% even with 10 or more times of iteration, an index of convergence of smoothing was introduced and used to determine appropriate iteration times. RESULTS: For solid neoplasms, appropriate times of iteration was three. With leukemia, the proportion of cells left out of their confidence intervals remained within 5% with iteration 27 times. Therefore, an index of convergence for the smoothing process was used to avoid over-smoothing. As a result, a value of five was obtained. In the case of car traffic accidents, even only a single smoothing did not fulfill the primary condition, because mortality rates increased very rapidly from age classes of 10-14 to 15-19, the mortality rates for the latter being about ten times larger. To deal with this, the proportion of cells left out of their confidence intervals was calculated excluding these age classes. As a result, five times was found to be appropriate. CONCLUSION: The smoothing method in this study is simple and easy to apply to mortality data. And procedure to determine appropriate iteration times is based on an inferential statistical method. Applying the method to three kinds of mortality data, it was found to be practical and useful for clarification of their characteristics and trends. PMID- 11433741 TI - [Trends in the incidence of cardiovascular diseases and risk factors among urban and rural Japanese males]. AB - OBJECTIVES: To examine long-term trends in the incidence of coronary heart disease, stroke, and their risk factors among Japanese populations, we explored 32 years of surveillance data for male residents in urban and rural areas in Japan. METHODS: The surveyed populations were 40-79 year-old male residents in M community (population over 40 years old in 1995 was 11,121) of Y City in Osaka (urban area; Osaka) and I town (n = 3,571) in Akita prefecture (rural area; Akita). Incidence rates of coronary heart disease (myocardial infarction, angina pectoris), sudden cardiac death, percutaneous transluminal coronary angioplasty (PTCA) and stroke per 1,000 person-years were calculated for 1964-71, 1972-79, 1980-87 and 1988-95. Risk factors were evaluated by cross-sectional surveys conducted in the median years for each period. Dietary intake was examined by the 24-hour recall method in the latter three periods. RESULTS: Age-adjusted incidence of coronary heart disease per 1,000 men increased progressively from 0.27 in 1964-71 to 0.90 in 1988-95 (P = 0.222 for trend) among 40-59 year old residents in Osaka. Among their 60-79 year old counterparts, though the incidence was 2.62-3.11 and did not change over the periods studied, the combined rates for coronary heart disease and men who had a PTCA reached 3.79 in 1988-95. In contrast, the incidence of coronary heart disease among Akita residents did not change over time and stroke declined 70 percent between 1964-71 and 1988-95 (P < 0.001) in both 40-59 and 60-79 year age group: The decrease in cerebral infarction was less marked between 1980-87 and 1988-95 among 60-79 year old individuals. Significant increases in diastolic blood pressure, total serum cholesterol, body mass index, salt intake and total fat percent of total energy in Osaka, were associated with the elevation in the incidence of coronary heart disease. In Akita, blood pressure leveled off over the study period, but decreased less during the last decade whereas the prevalence of obesity increased. CONCLUSION: Trends in cardiovascular diseases and their risk factors differ among different geographical areas. The present long-term study, in particular, demonstrated an increase in the incidence of coronary heart disease among urban Japanese male residents in Osaka between the 1960s and the 1990s. PMID- 11433742 TI - [A study of the psychological factors underlying weight-loss behavior among female adolescents]. AB - PURPOSE: The purpose of present study was to investigate to the relationship between tendency to diet and psychological factors in female adolescents who were not obese. METHODS: The Japanese Decisional Balance Inventory (KC-DBI), Japanese Situational Appetite Efficacy (KC-SAM) and Japanese Diet Efficacy Measure-6 (KC DEM6) were assessed. The participants were 853 female college students who were divided into underweight and normal groups with reference to their BMI. RESULTS: The results showed that score for KC-DBI, KC-SAM (negative feeling, hunger, relaxation) and KC-DEM6 differed with the degree of appetite change. A main effect on body figures was indicated only for KC-SAM (relaxation). There were no interactions in this analysis. Active participants showed a tendency to consider weight loss as a merit and to have fewer difficulties in controlling urges to overeat when they were feeling negative than the others. Furthermore, they had little efficacy in controlling their desire to be thin. We suggest that discussion of the merits of weight loss and interventions to enhance eating behavior are effective for health promotion in female adolescents. DISCUSSION: This study indicates some suggestions for educating female adolescents about the problems related to inappropriate dieting, and the results can used in the development of intervention programs. PMID- 11433743 TI - [A proposal for a new definition of excess mortality associated with influenza epidemics and its estimation]. AB - PURPOSE: As methods for estimating excess mortality associated with influenza epidemic, the Serfling's cyclical regression model and the Kawai and Fukutomi model with seasonal indices have been proposed. Excess mortality under the old definition (i.e., the number of deaths actually recorded in excess of the number expected on the basis of past seasonal experience) covers the random error for that portion of variation regarded as due to chance. In addition, it disregards the range of random variation of mortality with the season. In this paper, we propose a new definition of excess mortality associated with influenza-epidemics and a new estimation method, considering these questions with the Kawai and Fukutomi method. RESULTS: The new definition of excess mortality and a novel method for its estimation were generated as follows. Factors bringing about variation in mortality in months with influenza-epidemics may be divided into two groups: 1. Influenza itself, 2. others (practically random variation). The range of variation of mortality due to the latter (normal range) can be estimated from the range for months in the absence of influenza-epidemics. Excess mortality is defined as death over the normal range. CONCLUSION: A new definition of excess mortality associated with influenza-epidemics and an estimation method are proposed. The new method considers variation in mortality in months in the absence of influenza-epidemics. Consequently, it provides reasonable estimates of excess mortality by separating the portion of random variation. Further, it is a characteristic that the proposed estimate can be used as a criterion of statistical significance test. PMID- 11433744 TI - [Survey of consciousness of community residents regarding the community life of disabled persons with service dogs]. AB - PURPOSE: Service dogs have received much attention as effective choices for promoting participation in society of disabled persons. The purpose of this study was to obtain the fundamental data to prepare guidelines regarding service dogs use in community life, by making clear the problems in terms of public health and community welfare through a survey of attitudes towards service dogs in community residents. METHODS: Study 1 was conducted as follows. The subjects were 423 community residents between their teens and seventies who participated in health and welfare lectures in 4 metropolitan areas. A questionnaire survey was conducted just after watching a promotion video about service dogs, including several items on aspects not accept, were anxious about or were unpleasant in the case of community life of a disabled person and a service dog. Study 2 was as follows. A questionnaire survey was conducted inside a supermarket in the central part of Tokyo. One group of subjects were 103 shopping customers who witnessed a shopping scene featuring a disabled person with her service dog, and the other subjects were 84 customers who were not witnessed to the scene. The same survey was simultaneously performed for impression about the shopping scene. RESULTS: Study 1 showed that 31.8% of all the respondents replied that they had some knowledge of service dogs before watching the video. A total of 20.4% (32.5% of the elderly) replied that there were places where service dogs should not enter. Concretely speaking, these included restaurants for 7.3-22.5% and medical institutions for 8.5-12.5% of the respondents. A third of all the respondents (51.3% of older persons) replied they could not endure some actions by service dogs, concretely "carrying food in the mouth" was highly pointed out by 27.3%. Zoonosis (15.3-19.0%), hair scattering (9.5-21.3%), injuries (4.4-7.4%) were also pointed out as main sources of anxiety or uncomfortable feelings. Such negative opinions were especially apparent among those aged 60 years and older. Also, 44.1% knew some disabled person. Cross tabulation showed those who had experience of taking care of a dog were significant less anxious about "vague anxiety or discomfort," and "zoonosis" than those who had not (P < 0.01). On the other hand, most of both witness (79.6%), and non-witness groups (81.0%) replied that it was heartwarming that the service dog assisted with shopping. None expressed discomfort about the presence of "service dog" inside the supermarket. In terms of carrying a commodity in the mouth, 92.2% and 89.1% in both groups replied that they don't mind, especially if safety was guaranteed for public health, and 91.3% and 95.2% in both groups replied that they had no other problems. Moreover, none of them would not entering stores. Significant differences were seldom in replies between the witness and non-witness groups. CONCLUSION: Although the respondents were extremely affirmative regarding the image of service dogs, it became clear that anxiety about public health aspects such as "zoonosis" or "utilization of medical institutions" was persisting. On the whole, negative opinions were more apparent in the elderly. PMID- 11433745 TI - [Usefulness of the extended ADL scale for elderly community residents]. PMID- 11433746 TI - [The process taken for the official announcement of "Toward smoke-free society by Japanese society of public health]. PMID- 11433747 TI - [Efficacy of contrast-enhanced color Doppler ultrasonography for the diagnosis of renal mass lesions]. AB - We evaluated the efficacy of contrast-enhanced color Doppler ultrasonography with a galactose-based echo contrast agent, Levovist, for the diagnosis of renal mass lesions. The final pathologic diagnosis of eight renal mass lesions was renal cell carcinoma in 6 patients, transitional cell carcinoma in 1 patient and renal cyst in 1 patient (age range, 47 to 77 years; mean, 59.1). Especially in patients with renal cell carcinoma, contrast-enhanced color Doppler ultrasonography showed enhancement of mass lesion blood flow signal intensities. In all patients, contrast enhancement was demonstrated. Diagnostic capacity was improved in all patients, excluding 2 in whom mass lesion blood flow was detected by plain color Doppler ultrasonography. Furthermore, in patients with transitional cell carcinoma, the hemodynamics of mass lesions could be visualized, and diagnostic capacity was high. Contrast-enhanced color Doppler ultrasonography with Levovist, showed markedly higher mass lesion blood flow than that obtained by the plain color Doppler procedure, improving diagnostic capacity. This procedure was useful for diagnosing renal mass lesions. PMID- 11433748 TI - [Enucleation of renal tumor using a microwave tissue coagulator]. AB - We report our clinical findings on 7 tumors in 5 patients who underwent successful enucleation of renal tumor with a microwave tissue coagulator. All patients presented with incidentally-found renal tumor on ultrasonography or computed tomography. One patient had bilateral renal tumors. The mean size of the renal tumor was 2.5 cm (Range 1.0-5.2 cm). One patient had a tumor larger than 5 cm, and was found to have renal insufficiency preoperatively. Four cases were clinically diagnosed as T1aN0M0. In all patients, the renal tumor was enucleated with a microwave tissue coagulator successfully without renal pedicle clamping. The mean operative time and blood loss were 241 minutes and 788 ml, respectively. The pathological diagnosis was renal cell carcinoma in all cases. Although 2 patients had preoperative renal insufficiency, there were no serious postoperative complications and no evidence of loss of renal function as a result of this operation. Our findings suggest that renal tumors can be enucleated with a microwave tissue coagulator safely and successfully without damaging renal function. PMID- 11433749 TI - [Availability of NMP22 (nuclear matrix protein 22) for the diagnosis of urothelial cancer]. AB - We evaluated the availability of NMP22 for the diagnosis of urothelial cancer. Of 154 patients with gross hematuria or microscopic hematuria, 14 patients with bladder cancer and 4 patients with upper urothelial cancer were detected. Of 73 patients continued to be followed for urothelial cancers, 11 patients with bladder cancer were detected. NMP22 was not available when a large number of RBC was present in urine. Also, NMP22 was significantly higher in patients with urothelial cancers. Interpretation of the ROC curve indicated an optimal cutoff value of 9.2 u/ml. Using 9.2 u/ml as a cutoff, the sensitivity and specificity were, respectively 87.0% and 64.0% in patients with hematuria, and 81.8% and 78.7% in patients with a history of TCC. This indicated that when NMP22 was used for the diagnosis of urothelial cancers, 12.0 u/ml, the value recommended as a cutoff in Japan, might be too high to differentiate patients with urothelial cancers. PMID- 11433750 TI - [Screening methods in the detection of bladder cancer: comparison of nuclear matrix protein-22, bladder tumor antigen and cytological examinations]. AB - We evaluated the utility of urinary parameters (Nuclear Matrix Protein-22: NMP 22, Bladder Tumor Antigen: BTA, and cytological examinations) for the diagnosis or post-therapeutic monitoring of bladder cancer. Thirty one tumor-bearing cases including 19 fresh cases and 40 tumor-free cases, were subjects of this study. Using identical voided urine samples, NMP-22, BTA and urinary cytology were examined. The mean values of NMP-22 (cut-off value is 12 U/ml) was 100.5 +/- 26.5 U/ml in the tumor-bearing group and 21.9 +/- 7.8 U/ml in the tumor-free group (p < 0.05): Sensitivity was 74.2%, and specificity was 67.5%. Sensitivity of BTA was 58.1%, and specificity was 97.5%. Only five cases were judged positive by urinary cytology: 16.1% in sensitivity and 100% in specificity. Thus, NMP-22 and BTA were more sensitive than urinary cytology. In conclusion, the new urinary parameters, NMP-22 and BTA, would be less invasive and useful as tumor markers of bladder cancer. NMP-22 seems suitable for screening before the diagnosis and BTA for the post-therapeutic follow-up study. PMID- 11433751 TI - [Prophylactic effect of pirarubicin (THP) on postoperative recurrence of superficial bladder cancer in terms of intravesical retention time]. AB - In order to determine the modality of prophylactic intravesical instillation of pirarubicin (THP = tetrahydropyranyladriamycin) following transurethral resection (TUR) of superficial bladder cancer, a prospective randomized study was performed. A total of 79 patients were randomized into "2-hour instillation" (A), "5-min instillation" (B) and "control" (C) groups. Prophylactic efficacy and side effects were analyzed in each group. In groups A and B, 20 mg of THP was first dissolved in 10 ml of distilled water, adjusted to 40 ml with saline and was administered intravesically once a week for 10 weeks, starting from 1 week after TUR. The recurrence-free rate was calculated in 65 evaluable patients. The one year recurrence-free rate was 70.2% in group A, 62.8% in group B and 52.1% in group C. The one-year recurrence-free rate was significantly higher in group A than in group C. Adverse effects were observed in 21.4% of the patients in group A and 40.7% in group B. There was no significant difference in the occurrence rate of side effects between these two groups. Taking the prophylactic efficacy and side effects into consideration, "2-hour instillation" seemed to be better than "5-min instillation". PMID- 11433752 TI - [A case of retroperitoneal fibrosis responding to steroid therapy]. AB - A 73-year-old man presented with acute renal failure due to bilateral hydronephroses. Urography and CT scan revealed stenosis and medial deviation of the bilateral ureters surrounded by a soft tissue mass at the level of the 4-5th lumber vertebrae. Under a diagnosis of retroperitoneal fibrosis, bilateral nephrostomies were temporarily placed and prednisolone administration was initiated at a dose of 20 mg/day. Ureteral passage markedly improved within 2 weeks in association with a decrease in size of the retroperitoneal soft tissue mass. The steroid therapy was continued for 4 months with decreasing dosing schedule. The patient has been doing well without any signs of recurrence for 3 months after the cessation of the steroid therapy. PMID- 11433753 TI - [Usefulness of power Doppler ultrasound in a patient with renal cell carcinoma in the wall of a simple renal cyst]. AB - We present a case of renal cell carcinoma in the wall of a simple renal cyst. A 54-year-old man visited our hospital with a complaint of right back pain. Ultrasound study revealed right hydronephrosis, a ureteral stone, and a 9 x 8 cm renal cyst, with a tumor, 2 cm in diameter, in the cyst wall. Power Doppler Ultrasound and enhanced computed tomography demonstrated hypervascularity of the tumor. Clinically, he was diagnosed as renal cell carcinoma and nephrectomy was performed. Pathological findings of the tumor showed clear cell carcinoma in the wall of a simple renal cyst. Twelve months after the nephrectomy, he was free from recurrence. PMID- 11433754 TI - [Experience of direct hemoperfusion using polymyxin B-immobilized fiber on patients with endotoxin shock from urosepsis]. AB - We report the effects of direct hemoperfusion using polymyxin B-immobilized fiber (PMX treatment) on two patients with endotoxin shock from urosepsis. In both cases, urosepsis caused by pyelonephritis from a ureteral stone progressed to endotoxin shock and disseminating intravascular coagulation (DIC). Single J catheter was indwelt in the ureter for the purpose of drainage before PMX treatment. Drainage of the origin of infection, using effective antibiotics and PMX treatment improved shock state and DIC earlier than we expected. PMID- 11433755 TI - [Complete remission of brain metastases from prostate cancer by gamma knife radiosurgery: a case report]. AB - A 59-year-old male visited us with a chief complaint of dysuria. The serum prostate specific antigen (PSA) level was within normal limits, and intravenous pyelography and urethrocystography showed no abnormal findings. Because of his urinary retention, transurethral resection of prostate was performed under a clinical diagnosis of benign prostatic hyperplasia. The pathological diagnosis was poorly differentiated adenocarcinoma of the prostate. Not only combination hormone therapy with goserelin acetate and flutamide, but also intermittent arterial infusion chemotherapy with cisplatin (CDDP) and pirarubicin (THP) using a reservoir system was administered. Additionally total pelvic irradiation was delivered. Magnetic resonance imaging (MRI) demonstrated that his prostate was reduced to less than 50% in size and he had no difficulty in voiding. He suddenly developed dysarthria and hemiplegia 3 months later. MRI and computed tomography (CT) revealed multiple brain metastases. After the gamma knife radiosurgery, neurological findings disappeared and MRI showed dramatic shrinkage of metastatic brain tumors. Metastasis to the pancreas was recognized on CT and he died of multiple organ failure 30 months after his first visit. PMID- 11433756 TI - [Malignant lymphoma of the prostate: a case report]. AB - A case of a primary malignant lymphoma of the prostate is presented. An 82-year old man visited our hospital complaining of anal pain. Digital rectal examination revealed an enlarged prostate gland, which caused a rectal stricture. A computed tomographic scan and magnetic resonance imaging (MRI) showed a large mass arising from the prostate and protruding to the rectum. According to the Working Formulation, he was diagnosed with non-Hodgkin's lymphoma of B-cell origin, diffuse, mixed, small and large cell. The results of bone marrow puncture and imaging studies led to the diagnosis of primary malignant lymphoma of the prostate. Because of poor performance status deriving from severe anal pain, radiation therapy was performed to control the pain. After improvement of his performance status, he received combination chemotherapy consisting of cyclophosphamide, adriamycin, vincristine and predonisone (CHOP regimen). His prostate markedly diminished in size, but pneumonia developed. His respiratory condition rapidly deteriorated, and he died of respiratory failure about two and a half months after the onset of his illness. Malignant lymphoma involving the prostate, whether primary or secondary, is very rare. In our understanding, this case is thought to be the 28th clinical case of a malignant lymphoma of the prostate in Japan. PMID- 11433757 TI - [A case of traumatic rupture of the testis: usefulness of magnetic resonance imaging]. AB - Traumatic rupture of the testis is rare because of the protection afforded by surrounding structures. Moreover, it is difficult to accurately diagnose preoperatively. A 17-year-old man was referred to our department with the complaint of painful swelling of the left testis after being hit by a basketball. Although ultrasonography and computed tomography did not reveal the rupture of the tunica albuginea, we preoperatively diagnosed the rupture of the left testis by magnetic resonance imaging (MRI). We repaired the tunica albuginea and preserved the left testis. In this report, the advantages of MRI for preoperative diagnosis of traumatic rupture of the testis are discussed. In addition, previous cases with traumatic rupture of the testis in the Japanese literature are also reviewed. PMID- 11433758 TI - [Giant epidermal cyst in the perineum extending into the pelvic space in the patient with polycystic kidney disease: a case report]. AB - A 75-year-old man with autosomal dominant polycystic kidney disease was admitted to our hospital with the chief complaint of a giant mass in the left side of the perineum that had gradually developed in size during the previous 20 years. Palpation revealed a neonatal head-sized cystic mass with no tenderness. Magnetic resonance imaging (MRI) demonstrated a well-circumscribed homogeneous mass extended to the dorsal side of the bladder. It was excised surgically without any injury of rectum or urinary tract. The resected specimen was 23 x 18 x 12 cm in size and 1.2 kg in weight. The pathological diagnosis was epidermal cyst. Cases with an epidermal cyst extending into the pelvic space are extremely rare with few cases having been reported in the world. PMID- 11433759 TI - [Clinical effects of a 3-month formulation LH-RH agonist (Zoladex LA 10.8 mg depot) in patients with prostate cancer]. AB - Pharmacodynamics (PD), anti-tumor effects, safety and pharmacokinetics of a 3 month formulation of goserelin (Zoladex LA 10.8 mg depot: "10.8 mg depot") were investigated in a collaborative multicenter study. Study participants were 40 Japanese patients with prostate cancer comprising 20 untreated patients and 20 switch patients who had been receiving Zoladex 3.6 mg depot for 3 months or longer. Serum testosterone levels, serum LH levels, prostate-specific antigen (PSA) levels and drug concentrations were measured until 12 weeks after a single subcutaneous dose of 10.8 mg depot. Anti-tumor effects were evaluated by means of changes in the tumor lesions and the PSA levels at 12 weeks. After administration to the untreated patients, 10.8 mg depot reduced serum testosterone to the castrate range within 4 weeks and the reduction was maintained for up to 12 weeks. In the switch patients, serum testosterone suppression that had been produced by previous treatment with Zoladex 3.6 mg depot was maintained for up to 12 weeks following 10.8 mg depot administration. The anti-tumor effect at 12 weeks was 90.0% including partial response cases. The ratio of PSA normalization at 12 weeks was 75.0%. Fifty-seven adverse reactions were observed in 27 of the 40 patients (67.5%), but none were clinically significant. Although a disease flare presented as urinary retention in 1 of the untreated patients, all patients completed the study. Serum goserelin was detected up to 12 weeks after the administration of 10.8 mg depot. In conclusion a single dose of 10.8 mg depot showed a satisfactory PD-effect and brought about clinical efficacy persisting for at least 12 weeks and was well tolerated in patients with prostate cancer. PMID- 11433760 TI - [Roles of posterior parietal cortex in stereopsis: characteristics of responses of axis-orientation-selective neurons in monkey caudal intraparietal area]. AB - Patients with parietal lesions may fail to adjust the orientation of their hand to that of a target object, or may make errors in judging the orientation of a bar. This suggests that the parietal cortex has a function in the discrimination of the orientation of objects. In this study we investigated the responses of axis-orientation-selective neurons in caudal intraparietal (CIP) area to stereoscopic stimuli. Among the characteristics we investigated responses to the length and thickness of objects, sensitivity to binocular disparity, and position invariance in depth. Computer generated stereoscopic stimuli were presented to the monkey on a 70 inch screen. Most of the neurons responded better to long or narrow stimuli. All neurons which were orientation selective only in the frontal plane were not disparity sensitive. Most of the neurons which were orientation selective in the sagittal or horizontal plane were sensitive for binocular disparity. The majority of these neurons had wide receptive fields and their responses were position-invariant. These results suggest that the axis orientation-selective neurons in CIP area encode the orientation of the longitudinal axis of objects in 3-dimensional space. PMID- 11433761 TI - [Relationship between clinical activity and lymphocyte subsets in patients with Guillain-Barre syndrome]. AB - T lymphocytes are probably of pathogenic importance in Guillain-Barre syndrome (GBS). Blood samples from 31 patients with GBS and from 25 healthy controls were studied with flow cytometry. Subsets of the CD4 population were determined: CD4+CD29+ (helper-inducer) cells and CD4+CD45RA+ (naive or suppressor-inducer) cells. Recently, deviations of circulating CD8 populations have been noticed. Therefore, subsets of the CD8 population were also determined: CD8+CD11b dull (suppressor-effector) cells and CD8+ CD11b bright (cytotoxic-effector/NK) cells. During the active stage of GBS, significantly increased proportions of CD4+HLA DR+ (activated CD4+) cells and CD4+CD29+ cells were found compared to those in control samples (p < 0.0005, p < 0.05, respectively). The proportion of CD8+CD11b dull cells was significantly lower than that in the control samples (p < 0.005). Sequential analysis showed a decrease in the proportion of CD4+HLA-DR+ cells and CD4+CD29+ cells from the active to the convalescent stage (p < 0.05, p < 0.005, respectively). The proportion of CD8+CD11b dull cells was low in GBS patients with high disease severity. PMID- 11433762 TI - [Clinical features of autosomal dominant spinocerebellar ataxias genetically unclassified into any known triplet-repeat diseases]. AB - To clarify the clinical features of genetically unclassified autosomal dominant spinocerebellar ataxias (AD-SCAs) treated in our hospital, we retrospectively analyzed the clinical findings of patients who could not be molecularly classified into SCA types 1 through 3, 6 through 8, 12 and dentato-rubro-pallido luysian atrophy (DRPLA). The clinical findings of 19 patients from 15 different families which form 14 percent of all patients with different types of AD-SCAs (136 patients, 105 families) were collected. Based on these, 17 patients from 13 families showed late-onset (mean age at onset: 46 years old) and slowly progressive truncal and limb ataxias of cerebellar origin. Some of them showed gaze-evoked nystagmus, increased deep reflexes without Babinski sign, tremor, mildly decreased vibration sense and mild rectourinary disturbances. All but one were still ambulatory after 15 years of their mean disease duration. An earlier disease onset in successive generations was observed in 8 families. Brain MRIs revealed cerebellar atrophy predominantly in the anterior vermis in all the patients and mild brainstem atrophy in only 3 patients. Although the clinical phenotype manifested by most of the genetically unclassified AD-SCAs is similar to that of SCA6, whether the phenotype is caused by expansion of triplet repeats or some mutations in a single gene or different mutations in various genes is still unclarified. PMID- 11433763 TI - [A case of paraneoplastic cerebellar degeneration with resting tremor]. AB - A 65-year-old woman was operated for gastric adenocarcinoma in 1989. Six years later, peritonitis carcinomatosa, swelling of periaortic lymphnodes and high serum CA-125 were discovered. She received chemotherapy with 5-FU and cisplatin resulting in reduction of ascites. In September, 1998, the swelling of left supraclavicular lymphnodes and the elevation of serum CA-125 reappeared. Pathological diagnosis of supraclavicular lymphnodes was adenocarcinoma. Serum CA 125 was normalized by chemotherapy using cisplatin, farumorubicin and endoxan. However, unsteadiness appeared since December 10, 1998 followed by dysarthria and involuntary movement of neck and upper limbs. These symptoms progressed subacutely. The physical examination on admission revealed swelling of left suraclavicular lymphnodes, nystagmus on lateral gaze, saccadic eye movement on smooth pursuit and severe cerebellar ataxia. In addition, resting tremor of 3-4 Hz was observed at right hand, left wrist and neck which tended to increase amplitude by calculation. Similar movements were seen in the left first toe, though the frequency was lower. Brain MRI revealed mild cerebellar atrophy. She was diagnosed as paraneoplastic cerebellar degeneration (PCD) by serum anti Yo antibody and clinical course. The study of HLA showed positive link to A4 without A24. The primary focus of adenocarcinoma in cervical lymphnodes was suggested to be ovary rather than stomach due to the pattern of immunostaining for cytokeratin, CEA and CA125, although no carcinoma was found in ovarium clinically. The feature of this case is a PCD with resting tremor of frequency of 3-4 Hz and negative link to HLA-A24 in Japanese. PMID- 11433764 TI - [A case of acute sensory neuropathy associated with cytomegalovirus infection]. AB - We report a non-compromised patient with acute sensory neuropathy (ASN) developed following cytomegalovirus (CMV) hepatitis. A 67-year-old man admitted to a hospital because of acute hepatic dysfunction accompanying fever and skin eruption. One month later, when hepatic function normalized, numbness and clumsiness started acutely first in the right upper limb next to all the extremities. He found difficulty in walking in a couple of weeks. One month after the commencement of neurological illness, he was referred to us. On examination, he had sensory limb ataxia. His gait was wide-based, and Romberg sign was positive. Position sense was severely diminished in the extremities. Skin sensation was also attenuated distally, while no motor weakness was noted. Tendon reflexes were almost absent. Nerve conduction studies revealed absent sensory potentials in all but the left median nerve, in which amplitude was 5.5 microV with sensory conduction velocity of 40.7 m/s. Motor conduction studies, on the other hand, appeared normal except for a slight focal delay in the right ulnar nerve across the elbow. Mild increase in F-wave latencies was noted. A sural nerve specimen taken two months after the neurological onset showed a marked decrease in myelinated fiber density and active fiber degenerations accompanying axonal sproutings. Sjogren syndrome and paraneoplastic neuropathy were excluded serologically and by comprehensive imaging techniques. Although IgM anti-CMV antibody was not detected, serum IgG anti-CMV antibody was positive and significantly increased during the neurological illness. The intrathecal antibody synthesis of IgG anti-CMV antibody was suggested by a low serum/cerebrospinal fluid (CSF) antibody ratio and a high CSF IgG index. From these observations, it was strongly suggested that acute hepatitis and subsequent ASN were associated with CMV infection in this patient. Although some cases with post-infectious ASN have been previously reported, this is the first report of ASN preceded by CMV infection. PMID- 11433765 TI - [Adult Sandhoff disease presented as a motor neuron disease phenotype with slow progression]. AB - A 35-year old Japanese male with adult Sandhoff disease was described, who was presented as a motor neuron disease phenotype with slow progression. At the age of 15, he first noticed weakness in his thigh. At the age of 28, his upper and lower motor neuron disturbances were disclosed. He was diagnosed as atypical amyotrophic lateral sclerosis. At the age of 34, a slight sensory disturbance appeared in the lower extremities. When he was admitted to our hospital, he displayed marked atrophy and weakness in his quadriceps femoris muscles, but no signs of mental deterioration and cerebellar ataxia. Because of the atypical course of motor neuron disease, hexosaminidase activity in peripheral leukocytes was determined. The assay of total hexosaminidase, hexosaminidase A and hexosaminidase B activities demonstrated low levels of these activities (7-15% of controls), leading the diagnosis of Sandoff disease. He was a member of non consanguineous family, and the abnormal patterns of hexasaminidase activities were different between his father and mother. These data appear to show that he is a compound heterozygote in the locus of the hexosaminidase B gene. This is the first Japanese case of adult Sanhoff disease presented as a motor neuron disease phenotype. PMID- 11433766 TI - [An autopsy case of pure autonomic failure with pathological features of Parkinson's disease]. AB - Pure autonomic failure (PAF) is an uncommon disorder, characterized by autonomic failure without other neurological deficits. We report here an autopsy case of pure autonomic failure with pathological features of Parkinson's disease. At age 79, the patient developed slowly progressive severe orthostatic hypotension, followed by decreased sweating and constipation. On admission, his blood pressure dropped after standing (from 133/97 to 71/22 mmHg) without tachycardia. Neurological examination revealed neither parkinsonism, nor cerebellar symptoms. Supine plasma norepinephrine levels were low and intravenous norepinephrine administration showed denervation hypersensitivity. He was diagnosed as having PAF and treated by amezinium, midodrine, L-threodops, and fludrocortisone with elastic stockings but without any effects. He died of pneumonia about 2 years after the onset of the disease. Postmortem study revealed Lewy bodies in the nucleus basalis of Meynert, substantia nigra. Edinger-Westphal nucleus, locus ceruleus, and dorsal vagal nucleus. Neuronal loss was found in above-mentioned sites, except for Edinger-Westphal nucleus and intermediolateral nucleus. This is a rare and important report on a patient with PAF who satisfied the strict clinical criteria on PAF reported in 1996, and showed neuropathological findings, consistent with Parkinson's disease. PMID- 11433767 TI - [A case of neuro-Behcet's disease presenting with chorea]. AB - A 46-year-old man was admitted to our hospital because of emotional instability and involuntary movement of the right upper limb. Neurological examination revealed inability to concentrate, emotional incontinence, recent memory disturbance, chorea of bilateral upper limbs and neck, and bilateral pyramidal signs. Brain MRI showed atrophy of bilateral caudate nucleus and diffuse abnormal intensity area with low intensity on T1-weighted images and high intensity on T2 weighted images in cerebral white matter around the lateral ventricles. Huntington's disease was suspected at first, but it was ruled out by DNA analysis. After admission, oral and genital aphthae developed and the CSF examination showed pleocytosis (273 leukocytes/mm3; 39 polymorphonuclear leukocytes and 234 lymphocytes), so we diagnosed this case as neuro-Behcet's disease. Although basal ganglia is occasionally involved in neuro-Behcet's disease, chorea is rare. Neuro-Behcet's disease should be considered as a cause of chorea. PMID- 11433768 TI - [A case of primary Sjogren's syndrome with acute transverse myelopathy and polyneuropathy as the initial manifestations]. AB - We report a case of primary Sjogren's syndrome with acute transverse myelopathy and polyneuropathy as the initial manifestations. A 72-year-old man developed acute right hemiparesis and his symptoms deteriorated to quadriparesis in four days. A cervical spinal MRI showed an extensive intraparenchymal lesion with high T2-weighted signal intensity, gadolinium enhancement, and cord swelling. An electromyographic study and a sural nerve biopsy showed severe axonal degeneration. The patient complained of thirst and a salivary gland biopsy revealed inflammatory changes, while salivary gland scintigraphy showed diminished secretion. These findings led to a diagnosis of Sjogren's syndrome. Only one case of Sjogren's syndrome with acute transverse myelopathy as the initial manifestation has been reported. In our case, however, polyneuropathy was observed in addition to transverse myelopathy as the initial manifestation. Even if prominent sicca symptoms are absent, all patients with acute transverse myelopathy and polyneuropathy should be carefully examined with considerations of possible presence of Sjogren's syndrome. PMID- 11433769 TI - [A case with trigeminal herpes zoster manifesting a long lesion of the spinal trigeminal nucleus and tract on MR T2-weighted image]. AB - We reported a 53-year-old man with the right trigeminal herpes zoster with preceding neuralgia (preherpetic neuralgia) in the right upper cervical nerve area. He developed dysesthesia and scapular pain in the right second cervical nerve area. 5 days later, herpes zoster emerged in the area of the right maxillary division of trigeminal nerve. Furthermore, he developed paralysis on the right facial muscle on the 12th day after the onset of scapular pain. Neurological examination revealed decrease in superficial sensation accompanied by pain and dysesthesia in the areas innervated by the right maxillary division of trigeminal nerve and the right second cervical nerve, and the right peripheral facial nerve palsy. Any rash was not observed in the right second cervical nerve area throughout the course. The cerebrospinal fluid showed a mild mononuclear pleocytosis. The antibody titer for varicella zoster virus (VZV) was elevated in both cerebrospinal fluid and blood serum. T2-weighted magnetic resonance (MR) image revealed a continuously long high-signal lesion corresponding to the right spinal trigeminal nucleus and tract, extending from the lower pons to the second cervical segment of the spinal cord. This lesion could have resulted from a centripetal migration of VZV from the Gasser ganglion to the spinal trigeminal nucleus and tract, which was probably related to the preherpetic neuralgia in the upper cervical nerve area without rash. PMID- 11433770 TI - [A case of motor neuron disease with presenile dementia showing bilateral degeneration of the pyramidal tract on cranial MRI]. AB - A 58-year-old man developed dysarthria followed by a personality change. Subsequently, he developed muscle weakness and atrophy of the left upper and lower limbs, leading to repeated falls when he tried to walk. Neurological examination showed mild dementia, dysarthria, dysphagia, atrophy and fasciculation of the tongue, and muscle weakness and atrophy of all four extremities, particularly on the left side. Deep tendon reflexes were slightly diminished in the upper limbs and slightly exaggerated in the lower limbs without Babinski's sign. Cranial MRI revealed marked atrophy of the medial portions of the temporal lobes, more striking on the right, and T2-weighted imaging revealed symmetrical high-intensity signals from the posterior limbs of the internal capsules to the cerebral peduncles in the midbrain, extending to the pons on the left. 125I-IMP SPECT showed diffuse reduction of RI uptake in the frontal and temporal lobes, which was more marked on the right. We diagnosed this is a case of motor neuron disease with presenile dementia, which Mitsuyama et al. proposed as a new clinical entity, as well as a rare example of bilateral degeneration of the pyramidal tract on cranial MRI. PMID- 11433771 TI - [A family of early childhood-onset Charcot-Marie-Tooth disease type 2]. AB - We reported a man and his son with Charcot-Marie-Tooth disease (CMT) type 2. Their age at onset was about 5 years. Their clinical examinations revealed muscle atrophy and weakness of both distal lower limbs, foot-drop, a reduction of the reflex in both Achilles tendons and sensory impairment of the glove and stocking type. Nerve conduction studies revealed remarkably low amplitude of compound muscle action potential compared to conduction velocity. The nerve biopsy of their sural nerves revealed loss of large myelinated fiber. We presumed that the clinical features of their disease were compatible with CMT2A or 2B. DNA analysis of our family members performed with microsatellite markers linked to the candidate regions of CMT2A and 2B, did not show apparent positive results. We speculate that this family was a novel gene locus of CMT type 2. PMID- 11433772 TI - [Pure dysarthria due to pontine infarction]. AB - A 60-year-old man developed dysarthria in the morning. Neurological examination revealed no abnormality except for dysarthria. We diagnosed the patient as having a lacunar infarction. T2-weighted brain MRI revealed hyperintense lesion in the ventrolateral portion of the left upper pontine base. The clinical symptom completely disappeared on day 20. This is the first report on the MRI findings of the patient who showed pure dysarthria, which responsible lesion is in the pontine base. Involvement of corticopontine and corticobulbar tracts were considered to be responsible for the dysarthria in this case. This case may be useful for the somatotopy of corticopontine and corticobulbar tracts with articulation in the pontine base. PMID- 11433773 TI - [Formations and reactions of aromatic furazan compounds]. AB - A reaction from various kinds of nitroquinoline with hydroxylamine in potassium hydroxide alkalinity produced a novel product, furazanoquinoline, besides the known amino derivatives. The products obtained were furazano [3,4-f] quinoline (5) from 5-nitroquinoline (1) and 6-nitroquinoline (6), and furazano [3,4-h] quinoline (10) from 7-nitroquinoline (8) and 8-nitroquinoline (11). The reaction mechanism was believed to be as shown in Figs. 2 and 3. The photoreaction of benzofuroxan (19) in acetonitrile containing a little water, under a high pressure mercury lamp, produced 1H-azepine-2,7-dione (20), while under irradiation using a low pressure lamp, 6H-furazano [4,5-c] carbazole-3-oxide (21) and compound 20 were obtained. Then the photoproduct 20 produced photodimer 22 by irradiation in acetonitrile: water (9:1, v/v) using a high or low pressure mercury lamp, while photolysis with alkali proceeded as in the photoreaction of N alkylimide to give 7-hydroxy-1H-azepine-2-one (23). When pyrido [2,3-c] furoxan (24) was irradiated in acetonitrile containing a little water with a low pressure mercury lamp, 3-nitro-2-pyridone (25) was obtained. When compound 24 was irradiated in the presence of morpholine with a low pressure mercury lamp in an argon atmosphere, 6-morpholinopyridine 2,3-dioxime (26) was produced. Quinoxaline 1,4-dioxide derivatives (31, 33), phenazine 5,10-dioxide derivatives (36, 37) and pyrido [2,3-b] pyrazine derivatives (38, 39) were synthesized from the corresponding furoxan catalyzed by silica gel or molecular sieves, and their antibacterial properties were evaluated. The results of antibacterial screening tests in vitro, revealed strong activity against Bacteroides fragilis. PMID- 11433774 TI - [Development of catalytic reactions using water as a solvent]. AB - In recent years, organic reactions in aqueous media have attracted much attention, not only because these reactions eliminate the necessity of vigorous drying of solvents and substrates, but also because unique reactivity and selectivity are often observed in the aqueous reactions. Furthermore, organic reactions in water may lead to the development of environmentally friendly chemical processes. We have now developed various types of catalytic organic reactions in aqueous media. Catalytic asymmetric aldol reactions in aqueous media have attained by using rare earth metal triflates and a chiral multi-dentate ligand. Lewis acid catalysis has become feasible in water by using combinations of water-compatible Lewis acidic cations and anionic surfactants. These new catalysts were found to form stable colloidal dispersions and catalyze various reactions in water. Moreover, Bronsted acid- and palladium-catalyzed reactions in water have also been performed with the aid of surfactants. PMID- 11433775 TI - [Crystalline molecular complexes generated between surfactants and various additive compounds: an attempt to modify sparingly water soluble drugs to easily water soluble drugs]. AB - It has been established that ionic surfactants, whether cationic or anionic, provide crystalline molecular complexes stably with various aromatic compounds as additives. Among various additives phenolic compounds are especially favorable for both surfactant species. Other than these additives, cationic surfactants easily provide crystalline molecular complexes with basic materials and even with non-polar substances such as biphenyl and phenanthrene. The isolated surfactant molecular complexes were analyzed by X-ray. To date the number of the obtained surfactant molecular complexes exceeds two hundred species or more and thirty or more of them were structurally analyzed. These surfactant molecular complexes were shown to be novel surfactant species different from their mother species, supplied with enhanced solubility to the complex additives. We applied these characteristics to sparingly water soluble drugs as the complex additive to modify them into easily water soluble drugs. The obtained complex drugs revealed an improved behavior in solubility and dissolution rate compared with pure drugs. PMID- 11433776 TI - [Interactions between Asp-hemolysin from Aspergillus fumigatus and blood plasma components]. AB - Asp-hemolysin is a cytolytic toxin that is produced by Aspergillus fumigatus. This toxin is lytic for erythrocytes of humans, rabbits and sheep. However, Asp hemolysin is inactivated by the addition of serum or blood plasma. This study was undertaken to identify plasma components inhibitory to the hemolytic activity of Asp-hemolysin. alpha 2-Macroglobulin (alpha 2M) was isolated from the human blood plasma by affinity chromatography on a column containing Asp-hemolysin coupled to Sepharose. However, the hemolytic activity was only partially inhibited by alpha 2M. Apolipoprotein B (apoB)-containing lipoproteins, such as low density lipoprotein (LDL), inhibit the activity of this hemolytic toxin. When 20 micrograms apoB was added, the hemolytic activity was almost completely inhibited. Furthermore, similar inhibition was observed in the filtrates separated from the incubation mixture of Asp-hemolysin with LDL or apoB following ultrafiltration through a membrane with a molecular mass cutoff of 100,000. These results suggest that the inhibition by LDL is due to apoB binding to Asp hemolysin. The binding activity of LDL to Asp-hemolysin was measured. LDL binds to Asp-hemolysin with an affinity as high as the LDL receptor. The apparent Kd, determined by Scatchard plot analysis, was 8.9 x 10(-9) M 125I-LDL. Oxidized LDL (Ox-LDL), but not acetylated LDL, inhibited the hemolytic activity of this toxin. The inhibitory effects of Ox-LDL increased with the time of Cu(2+)-induced LDL oxidation. Similar inhibition was observed in the filtrate separated from the incubation mixture of Asp-hemolysin with Ox-LDL (for 2 h of oxidation) following ultrafiltration through a membrane with a molecular mass cutoff of 100,000. However, at longer LDL oxidation times, the inhibition by the filtrates was less than the control mixture without ultrafiltration. These results suggest that the inhibition of the hemolytic activity by Ox-LDL was due to the binding of Ox-LDL to Asp-hemolysin at short LDL oxidation times. PMID- 11433777 TI - [Pharmaceutical investigation of vancomycin hydrochloride eye drops and their topical application to MRSA eye infection]. AB - Two formulations of 0.5% vancomycin hydrochloride (VM) eye drops (VM-B and VM-C eye drops) were prepared by dissolving commercial VM powder for injection with preserved water B (PWB) containing phosphate buffer and preserved water C (PWC) containing only antimicrobial preservative, respectively. The VM-B eye drops have neutral pH (about 6.3), and the VM-C eye drops acidic pH of about 3.5. The pharmaceutical examination of these eye drops was performed regarding its clinical application to MRSA eye infection. In an irritability test using a rabbit's eye, the average number of winks after instillation of one drop of VM-B eye drops was 0.8 times/min and significantly smaller than that of VM-C eye drops (2.0 times/min). In dark storage at 4 degrees C, no change of VM concentration in both eye drops was observed for 25 weeks after preparation and the mean residual concentrations as determined by the HPLC-UV (240 nm) method were constant over 90% for 8 weeks, of the initial concentration. However, the residual VM concentration of VM-B eye drops under a room condition declined to 58% after 4 weeks and 20% after 8 weeks, and VM in light storage at 40 degrees C was not detectable after 8 weeks. The drug concentration of VM-C eye drops declined to 83% after 4 weeks and 74% after 8 weeks under a room condition, and to 46% after 4 weeks and 20% after 8 weeks under light storage at 40 degrees C. Under these storage conditions, the precipitation of VM related crystals was observed in both the eye drops when the residual percentage of VM was lower than 80%. Judging from HPLC chromatograms of a solution of the precipitated crystals, it was suggested that this crystal was degradation products of VM. The VM-B eye drops was applied to a patient with MRSA eye infection, because other medication was not effective. After continuous instillation of a drop per times every hour to both eyes, MRSA in corneal culture turned out negative after one week, and the clinical condition was remarkably improved. On the basis of the result of eye-irritability, VM-B eye drops with neutral pH was suggested to be superior to acidic VM-C eye drops from a safety point of view. It was also indicated that VM-B eye drops can be effectively used for 8 weeks under dark storage at 4 degrees C for MRSA eye infection, which is a useful piece information for the proper usage of the VM eye drops. PMID- 11433778 TI - [Preparation of a gel-forming ointment base applicable to the recovery stage of bedsore and clinical evaluation of a treatment method with different ointment bases suitable to each stage of bedsore]. AB - A novel ointment base suitable for the treatment of bedsore at the recovery stage was developed by the use of hydroxypropyl methylcellulose (HM-HPMC) modified on the basis of the hydrophobicity. A considerable sustained release of drug (minocycline hydrochloride) formulated to the ointment (T50 of 170 min) was attained with a macrogol ointment (MO) mixed with the HM-HPMC and Carbopol (CP) of the formulating ratio of 3:7. It was also found that a change in the formulating ratio of HM-HPMC and CP lead to a change in the drug release rate. The water absorption property of the ointment base, required to absorb on exudative solution in applying to the bedsore treatment, was as high as that of an ointment base containing hydroxypropyl cellulose (HPC) and CP reported in our previous paper. We clinically evaluated the effectiveness of the bedsore treatment, in which different ointment bases were applied to patients at different stages of the bedsore. A total of 22 cases were divided into two categories for applying to the different treatments. One category comprised of 11 subjects was treated with a povidone-iodine sugargel, which was the conventional method in our hospital, while the other 11 subjects were treated by the use of the newly developed ointment bases in consideration for the different stages of the bedsore. In comparison of the clinical results with the healing index, we ascertained that the latter method was significantly more efficacious (p < 0.01 0.05) than the conventional one. The effectiveness was emphasized in treating the intractable bedsore classified into the grades IV and V. Therefore, we confirmed that the newly developed ointment base formulation is useful in treating bedsore at each stage. PMID- 11433779 TI - [Effect of gallic acid derivatives on secretion of Th1 cytokines and Th2 cytokines from anti CD3-stimulated spleen cells]. AB - As reported previously (Kosuge et al., Yakugaku Zasshi, 120, 408 (2000)), methyl gallate, a gallic acid derivative, which has been one of compounds isolated from extracts of Psidium geneus Myrtaceae, selectively suppresses Th2 cytokine secretion. In the present study, to examine more effective compounds than methyl gallate, the effects of various gallic acid derivatives on the secretion of helper T cell subtype specific cytokines from anti CD3-stimulated spleen cells were investigated. Ten micrograms/ml of methyl gallate and ethyl gallate remarkably suppressed the secretion of IL-4 and IL-5, Th2 cytokines, but did not suppress meaningfully the secretion of IFN-gamma, a Th1 cytokine. On the other hand, the other gallic acid derivatives suppressed the secretion of both IL-4 and IFN-gamma. Ten micrograms/ml of methyl gallate suppressed the secretion of IL-2, a Th1 cytokine, but the same concentration of ethyl gallate did not suppress it. In conclusion, it seemed that ethyl gallate was the most selective inhibitor for the secretion of Th2 cytokines among gallic acid derivatives used in this study. PMID- 11433780 TI - Occurrence of coring in insulin vials and possibility of rubber piece contamination by self-injection. AB - Coring is reported to occur because rubber pieces are shaved off from a rubber stopper when a needle is inserted into the rubber stopper of transfusion liquid formulation. We verified whether coring really occurs in insulin vials of self injecting patients. We collected insulin cartridges from 30 hospitalized patients and used the primary injection (trial injection), the secondary injection and the cartridge remaining preparation as samples. We observed the rubber pieces using a microscope and measured the shape, number of pieces. The occurrence rate of coring was 73% for the primary injection, 47% for the secondary injection and 97% for the cartridge remaining preparation. The rubber pieces in the primary injection and the secondary injection which went through the needle are mostly in aggregate shape and the rubber pieces in the cartridge remaining preparation which did not go through the needle are mostly in needle-like shape. A number of small rubber pieces are found in both the primary injection and the secondary injection, indicating a high possibility that rubber pieces may be injected under subcutaneous tissue. The coring is considered to occur because needles are repeatedly inserted and rotated at the same spot. It is required to improve the structure to mount a needle to the pen-type injector in future. Coring is a very serious problem from the medical and pharmaceutical points of view. Further study should be made on the implication to latex allergy and lipodystrophy. PMID- 11433781 TI - Visual awareness and the on-line modification of action. AB - An influential theory of visually guided action proposes that (a) conscious perception of target displacement disrupts on-line action and (b) small target perturbations are inconsequential, provided the participant is unaware of them. This study examined these claims in a study of rapid aiming movements to targets. Novel features included on-line verbal reports of target displacement, and the factorial combination of small versus large displacements occurring near peak saccadic velocity or 100 ms later. Although awareness of target displacement had no effect on movement kinematics, even small target displacements near peak saccadic velocity affected kinematic measures. These results support both a strong view of visual stream separation in the on-line control of action and richer spatial coding by unconscious processes than has previously been acknowledged. PMID- 11433782 TI - Manipulating and recognizing virtual objects: where the action is. AB - In an earlier report (Harman, Humphrey, & Goodale, 1999), we demonstrated that observers who actively rotated three-dimensional novel objects on a computer screen later showed faster visual recognition of these objects than did observers who had passively viewed exactly the same sequence of images of these virtual objects. In Experiment 1 of the present study we showed that compared to passive viewing, active exploration of three-dimensional object structure led to faster performance on a "mental rotation" task involving the studied objects. In addition, we examined how much time observers concentrated on particular views during active exploration. As we found in the previous report, they spent most of their time looking at the "side" and "front" views ("plan" views) of the objects, rather than the three-quarter or intermediate views. This strong preference for the plan views of an object led us to examine the possibility in Experiment 2 that restricting the studied views in active exploration to either the plan views or the intermediate views would result in differential learning. We found that recognition of objects was faster after active exploration limited to plan views than after active exploration of intermediate views. Taken together, these experiments demonstrate (1) that active exploration facilitates learning of the three-dimensional structure of objects, and (2) that the superior performance following active exploration may be a direct result of the opportunity to spend more time on plan views of the object. PMID- 11433783 TI - The influence of stimulus properties on multisensory processing in the awake primate superior colliculus. AB - Multisensory integration is a process whereby information converges from different sensory modalities to produce a response that is different from that elicited by the individual modalities presented alone. A neural basis for multisensory integration has been identified within a variety of brain regions, but the most thoroughly examined model has been that of the superior colliculus (SC). Multisensory processing in the SC of anaesthetized animals has been shown to be dependent on the physical parameters of the individual stimuli presented (e.g., intensity, direction, velocity) as well as their spatial relationship. However, it is unknown whether these stimulus features are important, or evident, in the awake behaving animal. To address this question, we evaluated the influence of physical properties of sensory stimuli (visual intensity, direction, and velocity; auditory intensity and location) on sensory activity and multisensory integration of SC neurons in awake, behaving primates. Monkeys were trained to fixate a central visual fixation point while visual and/or auditory stimuli were presented in the periphery. Visual stimuli were always presented within the contralateral receptive field of the neuron whereas auditory stimuli were presented at either ipsi- or contralateral locations. Many of the SC neurons responsive to these sensory stimuli (n = 66/84; 76%) had stronger responses when the visual and auditory stimuli were combined at contralateral locations than when the auditory stimulus was located on the ipsilateral side. This trend was significant across the population of auditory-responsive neurons. In addition, some SC neurons (n = 31) were presented a battery of tests in which the quality of one stimulus of a pair was systematically manipulated. A small proportion of these neurons (n = 8/31; 26%) showed preferential responses to stimuli with specific physical properties, and these preferences were not significantly altered when multisensory stimulus combinations were presented. These data demonstrate that multisensory processing in the awake behaving primate is influenced by the spatial congruency of the stimuli as well as their individual physical properties. PMID- 11433784 TI - Failure to remap visuotactile space across the midline in the split-brain. AB - We examined the effect of posture change on the representation of visuotactile space in a split-brain patient using a cross-modal congruency task. Split-brain patient J.W. made speeded elevation discrimination responses (up versus down) to a series of tactile targets presented to the index finger or thumb of his right hand. We report congruency effects elicited by irrelevant visual distractors placed either close to, or far from, the stimulated hand. These cross-modal congruency effects followed the right hand as it moved within the right hemispace, but failed to do so when the hand crossed the midline into left hemispace. These results support recent claims that interhemispheric connections are required to maintain an accurate representation of visuotactile space. PMID- 11433785 TI - Electrophysiological evidence for the "missing link" in crossmodal attention. AB - Orienting attention involuntarily to the location of a sensory event influences responses to subsequent stimuli that appear in different modalities with one possible exception: orienting attention involuntarily to a sudden light sometimes fails to affect responses to subsequent sounds (e.g., Spence & Driver, 1997). Here we investigated the effects of involuntary attention to a brief flash on the processing of subsequent sounds in a design that eliminates stimulus-response compatibility effects and criterion shifts as confounding factors. In addition, the neural processes mediating crossmodal attention were studied by recording event-related brain potentials. Our data show that orienting attention to the location of a spatially nonpredictive visual cue modulates behavioural and neural responses to subsequent auditory targets when the stimulus onset asynchrony is short (between 100 and 300 ms). These findings are consistent with the hypothesis that involuntary shifts of attention are controlled by supramodal brain mechanisms rather than by modality-specific ones. PMID- 11433786 TI - Dissociating the neural correlates of item and context memory: an ERP study of face recognition. AB - We investigated the neural correlates of item and context retrieval using event related potentials (ERPs). Participants studied unfamiliar faces with happy or neutral expressions, and at test, they decided whether test faces were studied in the same or in a different expression, or were new. The parietal ERP effect, which is hypothesized to indirectly reflect medial-temporal lobe (MTL) function, was sensitive to item retrieval, whereas the frontal ERP effect, which is thought to reflect prefrontal cortex (PFC) function, was sensitive to context retrieval. Converging with lesion, functional neuroimaging (PET and fMRI), and ERP evidence, these results support the notion that item retrieval is primarily associated with MTL function whereas context retrieval is primarily associated by PFC function. PMID- 11433787 TI - Brain activations associated with shifts in response criterion on a recognition test. AB - Sensitivity and bias can be manipulated independently on a recognition test. The goal of this fMRI study was to determine whether neural activations associated with manipulations of a decision criterion would be anatomically distinct from neural activations associated with manipulations of memory strength and episodic retrieval. The results indicated that activations associated with shifting criteria (a manipulation of bias) were located in bilateral regions of the lateral cerebellum, lateral parietal lobe, and the dorsolateral prefrontal cortex extending from the supplementary motor area. These regions were anatomically distinct from activations in the prefrontal cortex produced during memory-based retrieval processes (manipulations of sensitivity), which tended to be more medial and anterior. These later activations are consistent with previous studies of episodic retrieval. Determining patterns of neural activations associated with decision-making processes relative to memory processes has important implications for Cognitive Neuroscience, including the use of these patterns to compare memory models in different paradigms. PMID- 11433788 TI - Is the donut in front of the car? An electrophysiological study examining spatial reference frame processing. AB - The process of describing an object's location relative to another object results in ambiguity. How do people handle this ambiguity? The present studies examined spatial language processing when use of different reference frames results in ambiguity. We investigated whether electrophysiological (ERP) measures of cognitive processing may elucidate underlying reference frame processing; in particular, we were interested in semantic integration. ERP results showed a larger N400, peaking between 300 and 375 ms, when the intrinsic frame was not used. Behavioural results mirrored this finding, indicating a reduced cognitive processing requirement for the intrinsic reference frame. Previous work has not definitively tied spatial reference frame processing to specific ERP components and their associated cognitive processes. Although the N400 peak seen in this data is early, additional work supports the N400 interpretation, thereby linking spatial frame processing to semantic integration. Results are discussed within the larger context of spatial reference frame processing. PMID- 11433789 TI - Do chimpanzees seek explanations? Preliminary comparative investigations. AB - During the past decade, considerable effort has been devoted to understanding whether chimpanzees reason about unobservable variables as explanations for observable events. With respect to physical causality, these investigations have explored chimpanzees' understanding of gravity, force, mass, shape, and so on. With respect to social causality, this research has focused on the question of whether they reason about mental states such as emotions, desires, and beliefs. In the studies reported here, we explored whether the chimpanzee's natural motivation for object exploration is modulated by a cognitive system that seeks explanations for unexpected events. We confronted both chimpanzees and young children with simple tasks which occasionally could not be made to work. We coded their reactions to determine if they appeared to be searching for an apparent cause (or explanation) of the task failure. The results of these preliminary studies point to both similarities and differences in how young children and chimpanzees react to such circumstances. PMID- 11433790 TI - Sensorimotor prediction and memory in object manipulation. AB - When people lift objects of different size but equal weight, they initially employ too much force for the large object and too little force for the small object. However, over repeated lifts of the two objects, they learn to suppress the size-weight association used to estimate force requirements and appropriately scale their lifting forces to the true and equal weights of the objects. Thus, sensorimotor memory from previous lifts comes to dominate visual size information in terms of force prediction. Here we ask whether this sensorimotor memory is transient, preserved only long enough to perform the task, or more stable. After completing an initial lift series in which they lifted equally weighted large and small objects in alternation, participants then repeated the lift series after delays of 15 minutes or 24 hours. In both cases, participants retained information about the weights of the objects and used this information to predict the appropriate fingertip forces. This preserved sensorimotor memory suggests that participants acquired internal models of the size-weight stimuli that could be used for later prediction. PMID- 11433791 TI - The role of vision in the on-line correction of illusion effects on action. AB - In this study, participants reached out and picked up a bar placed on a background grating that induced an illusion in the perceived orientation of the bar. The illusion had a large effect on the orientation of the hand early in the reaches, but this effect decreased continuously as the hand approached the target. This pattern occurred whether or not participants were allowed vision of the hand and target while reaching. These results are consistent with a "planning/control" model of action, in which actions are planned using a context dependent visual representation but monitored and corrected on-line using a context-independent visual representation. The hypothesized neural bases of these representations are discussed. PMID- 11433792 TI - Validity and efficiency of screening for history of depression by self-report. AB - Because of the recurrent nature of depression, there is a need for a rapid means of screening for history of depression that is either valid in itself or an efficient means of identifying respondents needing further assessment. This study examined the validity and efficiency of assessment of lifetime history of depression by self-report screening questions in comparison with the results of a structured interview assessment conducted a year earlier. Self-reported lifetime 2-week mood disturbance was unrelated to the results of the earlier interview. Self-report of treated episodes of mood disturbance were related to interview assessed history of depression, but too modestly for practical applications. Self report of past depression was more strongly related to concurrent distress than to the earlier interview assessment of history of depression. Implications of these findings for screening and assessment of history of depression are discussed. PMID- 11433793 TI - Refining the construct of psychopathy: towards a hierarchical model. AB - Psychopathy is characterized by diverse indicators. Clinical accounts have emphasized 3 distinct facets: interpersonal, affective, and behavioral. Research using the Psychopathy Checklist--Revised (PCL-R), however, has emphasized a 2 factor model. A review of the literature on the PCL-R and related measures of psychopathy, together with confirmatory factor analysis of PCL-R data from North American participants, indicates that the 2-factor model cannot be sustained. A 3 factor hierarchical model was developed in which a coherent superordinate factor, Psychopathy, is underpinned by 3 factors: Arrogant and Deceitful Interpersonal Style, Deficient Affective Experience, and Impulsive and Irresponsible Behavioral Style. The model was cross-validated on North American and Scottish PCL-R data, Psychopathy Screening Version data, and data derived from the Diagnostic and Statistical Manual of Mental Disorders (4th ed.; American Psychiatric Association, 1994) antisocial personality disorder field trial. PMID- 11433794 TI - Assessing readiness for change in the eating disorders: the psychometric properties of the readiness and motivation interview. AB - This study examined the psychometric properties of the Readiness and Motivation Interview (RMI), a symptom-specific measure of readiness and motivation for change in the eating disorders. For 4 symptom domains, the RMI assesses the extent to which individuals are in precontemplation, contemplation, and action/maintenance, and the extent to which change is made for internal versus external reasons. Ninety-nine individuals with eating disorders completed the RMI and measures to assess convergent, divergent, and criterion validity. RMI profiles revealed differences in readiness and motivation across symptom domains. The RMI demonstrated good reliability and construct validity, and RMI scores predicted anticipated difficulty of recovery activities, completion of recovery activities, decision to enroll in an intensive symptom-reduction program, and treatment dropout. The RMI may have important clinical applications by providing much-needed information on client readiness for action-oriented treatment. PMID- 11433795 TI - The adequacy of the MMPI-2 Harris-Lingoes subscales: a cross-cultural factor analytic study of Scales D, Hy, Pd, Pa, Sc, and Ma. AB - The authors investigated cross-culturally the factor structure of Scales D, Hy, Pd, Pa, Sc, and Ma of the Minnesota Multiphasic Personality Inventory--2 (S. R. Hathaway & J. C. McKinley, 1989) to examine the adequacy of the Harris-Lingoes (HL) subscales developed for these scales. A combined sample of 1,896 Israeli outpatients and inpatients and a sample of 1,020 American outpatients were used. Each scale was factor analyzed separately in the Israeli and U.S. samples and, within each sample, by gender. The results did not support the structural adequacy of the HL subscales for Scales D, Pd, Sc, and Ma but generally supported their suitability for Scales Hy and Pa. In addition, the results also suggested that all clinical scales share a common element of general distress akin to A. Tellegen's (1985) negative affectivity. These findings highlight the need for developing and validating a new set of subscales for most of the clinical scales, using external measures of relevant clinical and personality domains for which the current scales may serve as a basis. PMID- 11433797 TI - The Trail Making Test: a study in focal lesion patients. AB - The relationship of the Trail Making Test (TMT) to the frontal lobes was tested by comparing patients with damage to the frontal and nonfrontal regions to control participants. Although the analysis of time measurements, both raw and transformed, showed notable slowing of frontal groups, error analysis proved to be a more useful method of categorizing performance. Analysis of errors on Part B indicated that all patients who made more than 1 error had frontal lesions. Dividing the frontal damaged patients into subgroups on the basis of the number of errors yielded specificity of brain-behavior relations within the frontal lobes. Patients with damage in dorsolateral frontal areas were most impaired. Those with inferior medial damage to the frontal lobes were not significantly affected in TMT Part B performance. PMID- 11433796 TI - Confirmatory factor analysis of the Consumer Assessment of Health Plans Study (CAHPS) 1.0 Core Survey. AB - The National Consumer Assessment of Health Plans Study (CAHPS) Benchmarking Database was used to assess the factor structure and invariance of the CAHPS 1.0 Core Survey. Separate analyses were conducted with Latino and non-Latino Caucasian consumers drawn from commercial and Medicaid sectors (N = 15,092). Results demonstrated that the 23 CAHPS 1.0 report items measure consumer reports of experiences with 5 aspects of health plan performance: Access to Care, Timeliness of Care, Provider Communication, Health Plan Consumer Service, and Office Staff Helpfulness. Four items assessed global ratings of care. Analyses revealed an identical pattern of fixed and free factor loadings across all samples. Magnitude of factor loadings and correlations among factors was essentially equivalent within a common health service sector. A higher order factor analysis revealed that rating and reports of care showed marked convergence. PMID- 11433798 TI - Discrepancy between predicted and obtained WAIS-R IQ scores discriminates between traumatic brain injury and insufficient effort. AB - This study examines how well the discrepancy between predicted and obtained Wechsler Adult Intelligence Scale--Revised (WAIS-R) scores discriminate between insufficient effort (IE) and traumatic brain injury (TBI). The 27 IE patients performed significantly more poorly on the WAIS-R than the 48 moderate-severe TBI patients. Premorbid IQs were calculated with formulae that use demographics (Barona Index) or demographics and WAIS-R performance (Best-3 and the Oklahoma Premorbid Intelligence Estimation). Predictions were similar on the Barona, but IE patients' predicted IQs were lower than TBIs for measures with a performance component. IE patients demonstrated a greater discrepancy score (i.e., predicted IQ--obtained IQ) than TBIs; variable levels of sensitivity and specificity were obtained when discriminate functions were developed on these scores. The potential advantage of using discrepancy scores versus performance-based measures to detect insufficient effort is discussed. PMID- 11433799 TI - Criterion validity of the Holden Psychological Screening Inventory Social Symptomatology scale in a prison sample. AB - The 36-item, self-report Holden Psychological Screening Inventory (HPSI; R. R. Holden, 1996) and the Psychopathy Checklist--Revised (PCL-R; R. D. Hare, 1991) were administered to 214 male, adult prison inmates in Canadian federal correctional facilities. The 12-item HPSI Social Symptomatology scale, a measure of antisocial behavior, demonstrated a large effect size in significantly differentiating between PCL-R-identified psychopaths and nonpsychopaths. HPSI scales not theoretically related to psychopathic behavior showed no such significant effects. Findings are interpreted as supporting the criterion validity of the Social Symptomatology scale and suggest that this brief, self report screen has research and clinical merit. PMID- 11433800 TI - Effects of positive impression management on the NEO Personality Inventory- Revised in a clinical population. AB - Sixty adults in outpatient psychotherapy completed the NEO Personality Inventory- Revised (NEO PI-R, P. T. Costa & R. R. McCrae, 1992a). Half were instructed to fake good and half were given standard instructions. All completed the Interpersonal Adjective Scale--Revised, Big Five (J. S. Wiggins & P. D. Trapnell, 1997) under standard instructions, and their therapists completed the observer rating form of the NEO Five-Factor Inventory. A comparison group of 30 students completed the NEO PI-R under standard instructions. Standard and fake-good participants obtained significantly different NEO PI-R domain scores. Correlations between the NEO PI-R and criterion measures were significantly lower for faking than for standard patients. Validity scales for the NEO PI-R (J. A. Schinka, B. N. Kinder, & T. Kremer, 1997) were moderately accurate in discriminating faking from standard patients, but were only marginally accurate in discriminating faking patients from students. PMID- 11433801 TI - Reliable component analysis of the Stanford-Binet: Fourth Edition for 2- to 6 year-olds. AB - Reliable component analysis (RCA) was conducted on the Stanford-Binet: Fourth Edition subtests for 2- to 6-year-olds using the standardization sample. Scores were derived through RCA to assess the Verbal Comprehension and Non-Verbal Reasoning factors suggested for children in this age range. The scores derived through RCA had greater discriminant validity than did equally weighted scores, whose high intercorrelations preclude effective discrimination or incremental validity. The difference scores derived through RCA were compared with equally weighted difference scores in terms of reliability and three types of standard error. Differences between RCA scores were more reliable than were equally weighted differences. The more reliable differences resulted in more precise confidence intervals and more powerful significance tests. PMID- 11433802 TI - Discriminative validity of parent report of hypomanic and depressive symptoms on the General Behavior Inventory. AB - It often is difficult clinically to differentiate bipolar disorder from other mental health conditions in young people. This study evaluated a parent report measure of depressive and hypomanic/biphasic symptoms. Parents of 196 youths, who were 5 to 17 years old and presented at an outpatient research center, completed an adapted General Behavior Inventory (GBI). Factor analyses suggested two dimensions, depression (alpha = .97) and biphasic/hypomania (alpha = .95). Logistic regressions using these scales discriminated mood disorder versus disruptive behavior disorder or no diagnosis, unipolar versus bipolar disorder, and bipolar versus disruptive behavior disorder based on structured interviews. Classification rates exceeded 80%, and receiver operating characteristic analyses showed good diagnostic efficiency for the scales, with areas under the curve greater than .80. Results indicate that clinicians can use the parent-completed GBI to derive clinically meaningful information about mood disorders in youths. PMID- 11433803 TI - Development and validation of the Conflict in Adolescent Dating Relationships Inventory. AB - Four studies examined the Conflict in Adolescent Dating Relationships Inventory (CADRI), a measure of abusive behavior among adolescent dating partners. Exploratory factor analysis was used to refine items based on high school participants with dating experience (N = 393; 49% female). Confirmatory factor analysis was used to derive and cross-validate the factor structure with participants from 10 high schools (N = 1,019, 55% female; ages 14-16). The model structure fit for all grades and both sexes, with physical abuse, verbal abuse, and threatening behavior most representative of the underlying "abuse" factor. In Studies 3 and 4, the second-order abuse factor showed acceptable test-retest reliability, partner agreement, and correlation (significant for males only) between observer ratings of dating partners' interactions and youths' CADRI scores. Results support the CADRI as a measure of abusive behavior in adolescent dating relationships. PMID- 11433804 TI - [Relationship of Chlamydia pneumoniae infection to severity of coronary atherosclerosis in patients with chronic coronary artery disease and with normal coronary arteries]. AB - OBJECTIVES: Recent studies have demonstrated an association between infection with Chlamydia (C.) pneumoniae and coronary artery disease. However, the association is less clear in the Japanese population. The relationship of C. pneumoniae infection to severity of coronary atherosclerosis was investigated in patients with chronic coronary artery disease and with normal coronary arteries. METHODS: Serum levels of IgA and IgG antibodies to C. pneumoniae outer membrane complex were measured by enzyme-linked immunosorbent assay and C-reactive protein (CRP) analyses in 130 patients who underwent coronary angiography. Patients with unstable angina and recent myocardial infarction were excluded. Results were divided into three groups according to Gensini coronary score (GCS): normal (n = 19, GCS = 0); mild atherosclerosis (n = 56, GCS = 1-19); and severe atherosclerosis (n = 55, GCS > or = 20). RESULTS: Cut off indices of IgA and IgG in the atherosclerosis groups (severe: 1.53 +/- 0.72 and 1.67 +/- 0.97, mild: 1.58 +/- 0.92 and 1.42 +/- 0.86, respectively) were higher than in the normal group (1.22 +/- 0.59 and 1.28 +/- 0.82), but there were no significant differences. There were no correlations between indices of IgA and IgG, and GCS. The normal CRP group (n = 118, < 0.3 mg/dl) and the high CRP group (n = 12, > or = 0.3 mg/dl) showed no differences in IgA and IgG indices and GCS. CONCLUSIONS: Serum antibody indices against C. pneumoniae are not associated with the severity of coronary atherosclerosis in chronic stable coronary artery disease. PMID- 11433805 TI - Long-term outcome of patients with ergonovine induced coronary constriction not associated with ischemic electrocardiographic changes. AB - OBJECTIVES: This study investigated the long-term outcome of patients with coronary artery constriction induced with ergonovine but not associated with ischemic electrocardiographic changes. METHODS: The ergonovine provocation test was performed in 419 patients with suspected but unproven variant angina. Ergonovine maleate was administered into the coronary arteries at 8 micrograms/min for 5 min during cardiac catheterization. Patients were categorized into three groups according to their response to ergonovine. The positive group contained patients who developed coronary constriction of more than 90% in diameter associated with ischemic electrocardiographic changes and chest pain. The intermediate group contained patients who had coronary constriction of more than 90% but no electrocardiographic changes. The negative group contained patients who had neither significant coronary constriction nor ST segment changes. RESULTS: There were 305 patients, 49, and 65 in the negative, intermediate, and positive groups, respectively. Death occurred in six patients (2%), one (2%), and one (2%) in the negative, intermediate, and positive groups, respectively. Sudden cardiac death occurred in one patient in the negative group. Recurrence of chest pain with effectiveness of sublingual administration of nitroglycerine was observed in 26 patients (9%), 10 (20%), and 11 (17%) in the negative, intermediate, and positive groups, respectively. CONCLUSIONS: Some patients in the intermediate group might show "false negative" response to ergonovine so careful treatment with calcium antagonists should be continued in patients in the intermediate group as well as patients with vasospastic angina. PMID- 11433806 TI - [Clinical significance of measuring inferior vena cava dimension in patients with acute exacerbation of chronic heart failure]. AB - OBJECTIVES: This study determined the clinical significance of measuring the inferior vena cava dimension in patients with heart failure. METHODS: The subjects were 373 patients admitted due to acute exacerbation of chronic heart failure. The relationships were examined between inferior vena cava dimension in the end-expiratory phase on admission and in the stable state and etiology of heart failure, age, New York Heart Association (NYHA) functional classification in the stable state (before exacerbation), body weight, grade of tricuspid regurgitation, left ventricular end-diastolic dimension and left ventricular ejection fraction measured by two-dimensional echocardiography in the stable state and on admission, severity of lung congestion (Killip's classification) on admission, and clinical course during treatment. RESULTS: The inferior vena cava dimension was not correlated with etiology, age, NYHA functional classification, body weight, left ventricular end-diastolic dimension or left ventricular ejection fraction, but was correlated with the grade of tricuspid regurgitation (r = 0.78, p < 0.0001 in the stable state, r = 0.60, p < 0.0001 and on admission). The severity of lung congestion did not correlate with inferior vena cava dimension on admission or the increase in inferior vena cava dimension. The increase in body weight on admission was correlated with the increase in inferior vena cava dimension (r = 0.83, p < 0.0001), but did not correlate with inferior vena cava dimension on admission. Tricuspid regurgitation grade and inferior vena cava dimension were restored to the stable state when body weight reached stable state after treatment in patients (n = 202) with inferior vena cava dimension increased by over 5 mm on admission. CONCLUSIONS: Inferior vena cava dimension in patients with heart failure is correlated with the tricuspid regurgitation grade in the stable state and at acute exacerbation. The increase in inferior vena cava dimension between stable and exacerbated states, but not the actual value, is clinically useful to evaluate patients with acute exacerbation of chronic heart failure. PMID- 11433807 TI - Clinical manifestations of influenza a myocarditis during the influenza epidemic of winter 1998-1999. AB - OBJECTIVES: The clinical features of myocarditis that developed during the influenza epidemic of winter 1998-1999 were investigated to emphasize the need for medical attention to this disease. METHODS: Nine patients were treated under diagnoses of acute myocarditis during the winter of 1998-1999. Five (two males and three females, mean age 52 +/- 18 years) were examined with myocarditis associated with influenza A. The diagnosis of influenza A myocarditis was based on electrocardiographic and echocardiographic abnormalities, increased creatine kinase levels and at least a four-fold increase in influenza A virus titers using paired sera. RESULTS: All patients had preceding flu-like symptoms and fever. Cardiac involvement developed between 4 and 7 days after the onset of influenza symptoms. Dyspnea progressively worsened in three patients, one went into shock and one had persistent fever, cough and mild dyspnea without apparent cardiac symptoms. Three patients had ST elevation associated with Q waves and one had complete left bundle branch block. The creatine kinase levels were abnormally increased and global wall motion of the left ventricle on echocardiography was decreased in all patients. Two patients had diagnoses of fulminant myocarditis. One patient died of pneumonia following cerebral infarction, but the left ventricular dysfunction normalized in the remaining four patients. CONCLUSIONS: Cardiac involvement occurred between 4 and 7 days after the onset of influenza symptoms, and worsening dyspnea was the most common symptom. Electrocardiography, echocardiography and creatine kinase levels should be checked to determine the potential for cardiac involvement when patients present with suspected influenza associated with worsening dyspnea or prolonged weakness. Increasing the awareness of influenza myocarditis may help in the earlier identification and treatment of this disease during influenza epidemics. PMID- 11433808 TI - Quantification of left atrial appendage spontaneous echo contrast in patients with chronic nonalvular atrial fibrillation. AB - OBJECTIVES: The left atrial (LA) appendage is the most common site of thrombus formation in patients with atrial fibrillation, and integrated backscatter allows the quantiative assessment of LA spontaneous echo contrast (SEC). Integrated backscatter was used to examine the significance of measuring appendage SEC, specifically in relation to echocardiographic variables implying thromboembolism, in patients with chronic nonvalvular atrial fibrillation. METHODS: Fifty-two patients with chronic nonvalvular atrial fibrillation and no prior anticoagulant therapy (35 men, 17 women, mean age 66 +/- 7 years) underwent transesophageal echocardiography with integrated backscatter analysis. The LA and LA appendage integrated backscatter intensity were measured with the regions of interest placed in the LA cavity and the appendage, respectively. The integrated backscatter intensity values for these two chambers (corrected "LA" and "LA appendage" integrated backscatter intensity, respectively) were corrected using values from the left ventricular cavity. RESULTS: The LA appendage integrated backscatter intensity values were available in 44 patients (85%). Overall, the corrected LA appendage integrated backscatter intensity was significantly increased compared with the corrected LA integrated backscatter intensity (2.8 +/ 2.2 vs 2.0 +/- 1.8 dB, p < 0.001). The corrected LA appendage integrated backscatter intensity was inversely correlated with the LA appendage velocity (r = -0.37, p < 0.05), but not with the LA dimension, appendage size, or left ventricular function. The corrected appendage integrated backscatter intensity (4.5 +/- 2.3 vs 2.4 +/- 1.9 dB, p < 0.01) and LA integrated backscatter intensity (3.2 +/- 2.1 vs 1.7 +/- 1.7 dB, p < 0.05) were higher in patients who had LA appendage thrombus (n = 8) than those who did not. With the corrected appendage integrated backscatter intensity set at > or = 2.5 dB, the sensitivity for the presence of appendage thrombus was 88% and the specificity was 64%. CONCLUSIONS: Patients with chronic nonvalvular atrial fibrillation had a denser SEC in the LA appendage compared with SEC in the main LA cavity. The severity of the appendage SEC was influenced by the LA appendage function rather than its size. Quantification of SEC in the appendage, rather than main LA cavity, was more valuable for assessing embolic potential. PMID- 11433809 TI - [Effect of oxygen inhalation on myocardial opacification using levovist: an animal experimental study]. AB - OBJECTIVES: Levovist can opacify the myocardium via venous injection, and is widely used in the clinical field. However, left ventricular opacification deteriorates during oxygen inhalation using Albunex, one of the first generation of contrast agents. This study examines the effect of blood oxygen pressure on the myocardial opacification by Levovist. METHODS: Myocardial contrast echocardiography (Toshiba Power Vision 6000) was performed using second harmonic imaging, intermittent exposure of ultrasound (every 4 cardiac beats) and acoustic power of 1.4 mechanical index. The video intensity of the ventricular septum after venous injection of Levovist (0.5 ml of 300 mg/ml) was measured under various respiratory conditions by manipulating the respirator and oxygen inhalation. Arterial blood gas was measured and correlated with the myocardial video intensity. RESULTS: The video intensity of myocardial opacification was lower with higher oxygen pressure in the arterial blood. Visual recognition of myocardial opacification was only 14% under high oxygen pressure over 200 mmHg. Microbubbles of Levovist form from normal air, and are easily affected by the gas concentration of the surrounding blood. CONCLUSIONS: Myocardial opacification using Levovist is deteriorated under high oxygen pressure at examination. PMID- 11433810 TI - [Cerebellar infarction in a patient with para-scapular vascular murmur. Rendu Osler-Weber disease]. PMID- 11433811 TI - All-cause and cardiovascular mortality worldwide: lessons from geopathology. AB - Highly significant age-specific differences in all-cause and cardiovascular mortality rates are reported throughout the world. Male mortality is always higher than female mortality, but the sex ratio of mortality rates decreases with age. In most Western countries, all-cause mortality rates have been decreasing during the last 25 years. This decrease is almost uniquely due to a decrease in cardiovascular mortality. It is of crucial importance to try to determine the causes of these differences in mortality at the population level. Many factors may influence mortality: the level of medical care, genetic factors, nutrition, smoking habits, pollution, stress, socioeconomic factors, level of physical activity, etc. Of all the factors considered, nutritional habits and smoking combined with a high saturated fat intake appear to offer the most logical explanation for the existing differences in mortality. The intake of saturated fat has been linked to both cardiovascular and cancer mortality. Based on the modifications in the intercept and slope of the regression lines derived from the Gompertz equation (log mortality versus age) applied to a given population at a specific time, it has been possible to show that nutritional factors influence the aging process. The Gompertz equations point to the existence of a maximum age for the human race. Caution should be exercised when extrapolating population findings to individual subjects, in whom specific factors, e.g., genetic factors, may prevail. PMID- 11433812 TI - Doppler evaluation of systolic and diastolic heart failure in patients with cardiomyopathy. AB - BACKGROUND AND OBJECTIVES: Analysis of transmitral flow with Doppler echocardiography can currently evaluate left ventricular diastolic function noninvasively and easily. In addition, measurement of pulmonary venous flow and left atrial appendage flow clarifies not only left ventricular diastolic function but also left atrial systolic function in patients with left ventricular systolic failure and diastolic failure. METHODS AND RESULTS: Doppler echocardiography was used to investigate the differences in left ventricular diastolic dysfunction. In patients with systolic heart failure, left atrial dysfunction due to afterload mismatch to the left ventricle was related to the reduction of transmitral atrial filling wave in a restrictive pattern. In contrast, in patients with diastolic heart failure, transmitral flow seldom indicated a restrictive pattern because left atrial systolic function was preserved even with high pulmonary wedge pressure. CONCLUSIONS: Left ventricular diastolic dysfunction is somewhat distinct between patients with left ventricular systolic failure and patients with left ventricular diastolic failure. Atrial reversal flow in pulmonary vein has the potential to predict left ventricular filling pressure in patients with diastolic failure. PMID- 11433813 TI - Clinical significance of noninvasive assessment of left ventricular diastolic function by Doppler echocardiography. AB - Transmitral flow velocity pattern obtained by the pulsed Doppler technique reflects left ventricular (LV) diastolic function, but an increase in left atrial pressure pseudonormalizes the flow pattern and masks diastolic dysfunction. We propose an index to assess LV diastolic function using color M-mode Doppler echocardiography and the base-line shift technique. This index, flow propagation velocity, represents the average velocity of early diastolic LV filling flow from the mitral orifice to mid-ventricle. In patients with ischemic heart disease and dilated cardiomyopathy including those with pseudonormalized transmitral flow pattern, flow propagation velocity had a good correlation with the time constant of early diastolic LV pressure decay (Tau), indicating that flow propagation velocity is a useful noninvasive parameter of diastolic function which does not pseudonormalize. We also found a significant correlation between flow propagation velocity and Tau in hypertrophic cardiomyopathy patients in contrast to inadequate or no correlation between each of the conventional parameters and Tau. In addition, our recent data suggest that flow propagation velocity was distinctly decreased even in the patients with hypertension who did not show significant increase in LV mass index. Flow propagation velocity is a unique noninvasive parameter of LV diastolic function, which can sensitively and accurately detect the diastolic impairment in patients with different types of cardiac diseases with various loading conditions. PMID- 11433814 TI - Transthoracic Doppler echocardiographic assessment of left anterior descending coronary artery and intramyocardial small coronary artery flow in patients with hypertrophic cardiomyopathy. AB - OBJECTIVES: Transthoracic Doppler echocardiography with a high frequency transducer can visualize blood flow velocities in the left anterior descending coronary artery (LAD) and the intramyocardial small coronary artery in humans. However, the clinical utility for coronary circulation has not been established. To evaluate the reliability of transthoracic Doppler echocardiography for measuring coronary flow velocity, we firstly evaluated the coronary flow velocity and coronary flow reserve in the LAD in patients with coronary artery disease, and secondly evaluated the flow velocity in the intramyocardial small coronary artery in patients with hypertrophic cardiomyopathy. METHODS: Transthoracic Doppler echocardiographic LAD flow velocity was simultaneously recorded with Doppler guide wire before and after papaverin or adenosine triphosphate (ATP) infusion in 18 patients with coronary artery disease (mean age 61 years) during cardiac catheterization. Intramyocardial small coronary artery flow velocity by transthoracic Doppler echocardiography was examined in another 51 patients (mean age 53 years) with hypertrophic cardiomyopathy and 31 normal subjects (mean age 62 years). RESULTS: There were good correlations of coronary flow reserve measured by papaverin (r = 0.87) and ATP (r = 0.80) administration between the transthoracic Doppler echocardiography and Doppler flow wire methods in patients with coronary artery disease. Averaged diastolic peak flow velocity [77 +/- 36 (mean +/- SD) cm/sec] and time velocity integral (27 +/- 11 cm) in the small coronary artery in patients with hypertrophic cardiomyopathy were significantly higher (p < 0.05) than those in the LAD (46 +/- 20 cm/sec and 19 +/- 11 cm), respectively. However, there were no significant differences of flow velocities in the control subjects between averaged diastolic peak flow velocity (31 +/- 10 cm/sec) and time velocity integral (11 +/- 5 cm) in the small coronary artery and those in the LAD (32 +/- 11 cm/sec and 12 +/- 5 cm), respectively. Systolic reversal flow in the small coronary artery was observed in 41 patients with hypertrophic cardiomyopathy, but not in the controls. CONCLUSIONS: Transthoracic Doppler echocardiography using a high frequency transducer is a useful technique for noninvasive investigation of flow velocity dynamics in the LAD and intramyocardial small coronary artery in patients with hypertrophic cardiomyopathy. PMID- 11433815 TI - Clinical analysis of myocardial perfusion and metabolism in patients with hypertrophic cardiomyopathy by single photon emission tomography and positron emission tomography. AB - BACKGROUND AND OBJECTIVES: Changes in myocardial energy metabolism and their relation to coronary flow reserve in hypertrophic cardiomyopathy were assessed by myocardial fatty acid imaging with iodine-123 beta-methyliodophenyl pentadecanoic acid single photon emission tomography (123I-BMIPP SPECT) (fasting), glucose imaging with fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG PET) (fasting), and perfusion imaging with nitrogen-13 (13N)-ammonia PET (dipyridamole-stress and at rest) in adult patients with hypertrophic cardiomyopathy and with asymmetric septal hypertrophy. RESULTS: 123I-BMIPP defects mismatched with thallium-201 (201Tl) uptake were often observed in the hypertrophic septal regions indicating reduced fatty acid utilization incidence of 59% (22/37). 18F-FDG images showed diffusely increased uptake in most of the patients (73%, 27/37), but showed regionally increased 18F-FDG uptake at the septal regions in only two patients. Study of 18 hypertrophic cardiomyopathy patients with the mismatched 123I-BMIPP defects found that the severity of the defects correlated with reduced coronary flow reserve determined by the 13N ammonia PET study. On the other hand, changes in 18F-FDG images were not related to those in coronary flow reserve. CONCLUSIONS: These results suggest that the mismatched 123I-BMIPP defects, which indicate abnormality in myocardial fatty acid metabolism, occur under reduced coronary flow reserve, and may contribute to the prediction of progressive myocardial failure in patients with hypertrophic cardiomyopathy. The significance of 18F-FDG in patients with hypertrophic cardiomyopathy is still uncertain. PMID- 11433816 TI - Anthracycline-induced cardiomyopathies evaluated by tissue Doppler tracking system and strain rate imaging. AB - OBJECTIVES: Anthracycline cardiotoxicity is known to occur from the subendocardial side of the left ventricular wall. Recent advances of tissue Doppler echocardiography may allow the evaluation of anthracycline cardiotoxicity by dividing the left ventricular wall into the subendocardial half and subepicardial half. The present study assessed the feasibility using the tissue Doppler echo tracking system (M-mode) and myocardial strain rate imaging (B-mode) to noninvasively detect anthracycline cardiotoxicity. METHODS: The tissue Doppler echo tracking system (M-mode) was used to measure systolic thickening of the subendocardial layer (delta Endo), subepicardial layer (delta Epi), and whole wall (delta Total) of the left ventricular posterior wall in 41 normal subjects and three groups of patients receiving anthracycline: 34 patients in the low dose group, 19 in the middle dose group, and 12 in the high dose group. Strain rate is the spatial gradient of local velocities, reflecting local compression and expansion rates not affected by overall heart motion. Myocardial strain rate imaging (B-mode) was used in 25 normal subjects, 9 patients in the low dose group, and 10 patients in the high dose group. The ratio of peak systolic strain rate of subendocardium to that of subepicardium (peak strain rate endo/epi), and the ratio of integrated strain rate during ejection time of subendocardium to that of subepicardium (integrated strain rate endo/epi) were measured. RESULTS: Tissue Doppler echo tracking system (M-mode) measurement of delta Endo/delta Epi showed the most distinct difference and the least overlap of the data between normal subjects and patients, whereas delta Total failed to show significant differences. Myocardial strain rate imaging (B-mode) measurement of integrated strain rate endo/epi showed the most distinct difference and the least overlap of the data between normal subjects and patients, but ejection fraction failed to show statistically significant differences. CONCLUSIONS: These methods are highly sensitive tools for monitoring anthracycline cardiotoxicity. PMID- 11433817 TI - Hypertrophic cardiomyopathy in the elderly: significance of atrial fibrillation. AB - BACKGROUND AND OBJECTIVES: Atrial fibrillation (Af) is an important prognostic indicator for clinical deterioration in patients with hypertrophic cardiomyopathy (HCM), particularly in elderly patients, but its significance may vary from unselected to selected populations. This study investigated the risk of Af as a prognostic factor in long-term mortality and morbidity in Japanese patients with HCM at our institution. METHODS: All fatal and non-fatal cardiovascular events were estimated in 91 patients with HCM (mean age 51 +/- 14 years, 62 men) and follow up continued for 6.7 +/- 4.8 years. RESULTS: Either chronic or paroxysmal Af documented by standard electrocardiography (ECG) or ambulatory ECG monitoring at diagnosis and during follow-up were included with other clinical features in the survival analysis. Af was documented in 22 patients (24%). They were older than those without Af (57 +/- 10 vs 48 +/- 14 years, p < 0.005) and had larger left atrial dimensions (43 +/- 6 vs 36 +/- 5 mm, p < 0.01). Cardiovascular events occurred in 12 patients with Af (embolic events in five, heart failure in four, and sudden death in three), and in four patients without Af (embolic event in one, and sudden death in three). Death occurred in 10 patients (seven with Af and three without Af); sudden death in six patients (three with Af and three without Af) and four patients died of cerebrovascular embolism (four with Af and none without Af). Cox's regression analysis showed the occurrence of Af was the most useful predictor of cardiovascular death in patients with HCM. The event-free rate was significantly worse in patients with Af than in those without Af (p < 0.0001). CONCLUSIONS: Af is an important prognostic feature in patients with HCM, which occurs in older patients and in those with left atrial dilation. PMID- 11433818 TI - Molecular etiology of idiopathic cardiomyopathy in Asian populations. AB - BACKGROUND: Idiopathic cardiomyopathy was by definition a disease of unknown etiology and there are two major clinical forms, hypertrophic cardiomyopathy and dilated cardiomyopathy. Recent molecular genetic analyses have now revealed that mutations in genes for sarcomere cause hypertrophic cardiomyopathy leading to a hypothesis of hypertrophic cardiomyopathy as sarcomeropathy. On the other hand, mutations in genes for Z-disc component cause dilated cardiomyopathy speculating that dilated cardiomyopathy is cytoskeletopathy at least in part. METHODS: A large panel of Asian patients and families with hypertrophic cardiomyopathy or dilated cardiomyopathy was analyzed for gene abnormalities in all exons and adjacent introns of the known disease-related genes and in a part of several candidates of novel disease-related genes. RESULTS: Mutations in the genes for sarcomere were found in 47% of familial cases and 14% of sporadic cases of hypertrophic cardiomyopathy and there were locus and allelic differences in clinical phenotypes of hypertrophic cardiomyopathy patients. In contrast, only a few patients with dilated cardiomyopathy were identified for mutations in the known disease-causing genes. Mutations in the gene for titin, a giant molecule linking Z-disc with sarcomere components, were found in hypertrophic cardiomyopathy patients. CONCLUSIONS: The molecular etiologies of cardiomyopathy can be identified in about half of hypertrophic cardiomyopathy and a small part of dilated cardiomyopathy, suggesting that there are several novel disease causing genes. Identification of titin mutation in hypertrophic cardiomyopathy indicate that hypertrophic cardiomyopathy is in part considered as the cytoskeletopathy. PMID- 11433819 TI - Natural history of hypertrophic cardiomyopathy: Japanese experience. AB - BACKGROUND AND OBJECTIVES: Most patients with hypertrophic cardiomyopathy (HCM) remain clinically stable for long periods of time, whereas some patients progress to severe systolic dysfunction. Therefore, the natural history of HCM is largely unknown. METHODS: The present study followed up 59 patients with HCM (32 males, 27 females, mean age 38.6 +/- 13.6 years) for 10 years or more (mean 16.0 +/- 4.7 years) after the initial diagnosis. RESULTS: Eight of 17 patients who showed abnormal Q-waves at the initial examination had lost Q-waves, suggesting remodeling from asymmetric to generalized hypertrophy. The thickness of the interventricular septum showed remarkable changes, increasing by > or = 5 mm in 7 patients and decreasing by > or = 5 mm in 21. These observations indicate that ventricular remodeling occurs in patients with HCM. Follow-up electrocardiography demonstrated new Q-waves in 10 patients and bundle branch blocks or intraventricular conduction disturbances in 13. Left ventricular end-diastolic diameter increased from 41.6 to 48.1 mm, associated with a decrease in fractional shortening from 40.6% to 34.0%. Left ventricular systolic dysfunction, defined as left ventricular end-diastolic diameter > 55 mm or fractional shortening < 25%, developed in 13 patients. These observations indicate that myocardial disease including the conduction system is progressive in patients with HCM and finally deteriorates to systolic dysfunction. Left ventricular outflow obstruction also presented evolutional changes. At the initial study, 23 patients showed systolic anterior motion of the mitral valves. Systolic anterior motion disappeared in 13 patients, reduced in 2, increased in 2, and remained stable in only 6. One patient without systolic anterior motion at the initial study developed new systolic anterior motion. Impaired left ventricular filling increased left atrial diameter from 35.5 to 46.9 mm and atrial fibrillation frequently developed (24 patients). CONCLUSIONS: These findings suggest that HCM is a slowly progressive disease which develops evolutional remodeling of left ventricular hypertrophy and outflow obstruction, eventually progressing to systolic dysfunction with cavity dilation and wall thinning. PMID- 11433820 TI - Floppy mitral valve/mitral valve prolapse/mitral valvular regurgitation: effects on the circulation. AB - The floppy mitral valve prolapses into the left atrium in such a dynamic manner that the prolapsing floppy mitral valve becomes a space-occupying lesion within the left atrium. A significant result of the floppy mitral valve prolapsing into the left atrium during left ventricular systole is the development of a "third chamber" located between the mitral annulus and the prolapsing mitral valve leaflets. Since the blood in the third chamber does not contribute to forward stroke volume, the third chamber may have significant effects on stroke volume and cardiac output. The floppy mitral valve/mitral valve prolapse dynamics also affect left ventricular papillary muscle tension and traction, altering the patterns of left ventricular contraction and relaxation, activating papillary muscle and left ventricular stretch receptors, and contributing to the production of cardiac arrhythmias. Floppy mitral valve innervation patterns with distinct nerve terminals provide a neural basis for brain-heart interactions, augmented by mechanical stimuli from the prolapsing floppy mitral valve. With the onset of mitral valvular regurgitation, and gradual progression of the mitral valve regurgitation from mild, to moderate, to severe, alterations in left atrial and left ventricular chamber size and performance occur, resulting in left atrial and left ventricular myopathy. As a connective tissue disorder, floppy mitral valve/mitral valve prolapse may be associated with abnormal structural and elastic properties of the aorta, with resultant changes in aortic function. Progression of mitral valve regurgitation and the aging process also affect aortic function indices in an adverse manner. The phenomena associated with floppy mitral valve dysfunction, with prolapse of the mitral valve into the left atrium and the unique, resultant forms of mitral valve regurgitation, are dynamic in nature. As the long-term natural history of these interrelated phenomena is being clarified, it is apparent that the floppy mitral valve/mitral valve prolapse/mitral valve regurgitation influences the circulation in a global fashion. PMID- 11433821 TI - Thermal therapy for congestive heart failure: estimation by TEI index. AB - Systolic dysfunction commonly coexists with diastolic dysfunction in patients with congestive heart failure due to myocardial disease. Also, right ventricular dysfunction is frequently associated with left ventricular dysfunction. The Doppler total ejection isovolume (TEI) index, a combined measurement of systolic and diastolic myocardial performance, is more reflective of overall cardiac function than systolic or diastolic function alone in both ventricles, and provides a conceptually new measure of global cardiac function. Single sauna bathing improves acute hemodynamics and repeated sauna therapy improves cardiac function and clinical symptoms in patients with heart failure. More recently, improvement of vascular endothelial function were observed by repeated sauna therapy in patients with heart failure. The TEI index, combining systolic and diastolic myocardial function, is a promising tool for the quantitative assessment of global cardiac function before and after thermal therapy. PMID- 11433822 TI - Apical hypertrophic cardiomyopathy (apical hypertrophy): an overview. AB - Clinical and laboratory data of apical hypertrophy were reviewed based on our experience of over 200 consecutive patients, of whom 126 patients were followed up by myself for more than 1 year (1 to 29 years). Emphasis was placed on various aspects of electrocardiography including the natural course and "wax and wane" phenomenon of giant negative T waves (GNT). Recent diagnostic modality, i.e., cardiac magnetic resonance imaging was also stressed. Apical hypertrophic cardiomyopathy (apical hypertrophy) was mainly discovered by annual health check including electrocardiography and characterized by giant negative T waves (GNT; 1.0-(-)4.2 mV) in the left precordial leads (V4 or V5) in middle-aged men. Transition from normal T wave to negative T wave required several years and remained usually unchanged thereafter. This change may occur rather abruptly on rare occasions. Disappearance of GNT may also occur slowly and progressively in patients, in whom apical aneurysm had developed. The diagnosis may be obtained with echocardiography, left ventriculography or ultrafast computed tomography, but was most accurate with cardiac magnetic resonance imaging, by which identification of the diversity of hypertrophy was achieved, because the multiple short-axis views were accurately obtained in addition to the exact long-axis view. Hypertrophy was not simple but quite complex in both morphology and grade. Gene abnormality may be present even in cases of apical hypertrophy. The prognosis of apical hypertrophy in Japan has been benign, and heart failure due to atrial fibrillation and left ventricular aneurysm due to the destruction of hypertrophied muscle are thought to have prognostic importance, but these were rare in our series. PMID- 11433823 TI - Mitral annular calcifications and aortic plaques as predictors of increased cardiovascular mortality. AB - OBJECTIVES: Mitral annular calcifications (MAC) and aortic plaques are predictive for higher incidence of recurrent embolic events, the presence and severity of coronary artery disease and total cardiac deaths. However, no data exist comparing the impact of combined presence of both aortic plaques and MAC on embolic events, deaths associated with cerebral embolism or cardiac deaths during the follow-up. PATIENTS AND METHODS: In 100 consecutive patients with aortic plaques detected by transesophageal echocardiography (mean age 69.8 + 8.9 years, range 39-93 years, 49 males, 51 females) and in 49 age- and sex-matched control subjects (mean age 69.4 + 7.8 years, range 46-89 years, 27 males, 22 females) without aortic plaques (control group), the incidence of embolic events, deaths associated with embolic events and total cardiac deaths were evaluated during a mean follow-up of 32 +/- 9.7 months. The effect of the presence or absence of MAC on these parameters was also evaluated. RESULTS: Compared with the control group, patients with aortic plaques had a higher frequency of hypertension (p < 0.001), hypercholesterolemia (p < 0.01), smoking history (p < 0.01), MAC (p < 0.00001) and the presence of atherosclerotic changes in the carotid arteries (p < 0.05). During the follow-up, patients with aortic plaques had a higher incidence of stroke or transient ischemic attack (p < 0.0001), deaths associated with stroke (p < 0.005) and total cardiac deaths (p < 0.05) compared with the control group. Patients with both aortic plaques and presence of MAC had during the follow-up a significantly higher incidence of deaths associated with stroke (p < 0.0001, 16.0 times higher) and total cardiac deaths (p < 0.005, 4.7 times higher) compared with those without MAC and the presence of aortic plaques. Multivariant regression analysis showed MAC (p < 0.01) and hypercholesterolemia (p < 0.05) were the only independent predictors of stroke. MAC (p < 0.05) and age (p < 0.000001) were the only independent predictors of total cardiac death and stroke. Independent predictors of both aortic plaques and MAC were coronary artery disease (p < 0.00001), stroke (p < 0.005), hypertension (p < 0.001), female gender (p < 0.01) and hypercholesterolemia (p < 0.005). CONCLUSIONS: The findings of both aortic plaque and mitral annular calcification are highly predictive not only for strokes, but also for deaths associated with cerebral embolism and total cardiac deaths and are probably strong markers for higher severity of coronary artery disease. They are more predictive if both are present simultaneously compared to the presence of either MAC or aortic plaque alone. PMID- 11433824 TI - Mitral annulus dynamics: determinants of left ventricular filling. AB - OBJECTIVES: To examine the influence of mitral annular dynamics on left ventricular filling. METHODS: Twenty patients (mean age 64 +/- 6 years) with coronary artery disease and normal left ventricular function were studied by transesophageal echocardiography immediately before bypass grafting with the pericardium open. Pulmonary venous and transmitral velocities were recorded with pulsed wave Doppler. Mitral annulus position was derived by M-mode and its rate of change by digitisation of the records. From simultaneous high fidelity left atrial pressure, instantaneous power and its time integral work were derived. RESULTS: In all patients the power curve was triphasic, being positive in both early and late diastole and negative during ventricular systole. Systolic annular work was related to pulmonary venous peak systolic velocities (r = 0.74, p < 0.001). The ratio of the early to late diastolic annular work correlated with the transmitral E/A ratio (r = 0.82, p < 0.0001) and early diastolic work was related to pulmonary venous peak diastolic velocities (r = 0.74, p < 0.001). Pressure displacement curves of the annulus throughout the period of rapid ventricular filling were curvilinear with instantaneous stiffness increasing with distension. In addition all curves demonstrated a diastolic equilibrium point of zero stiffness. CONCLUSIONS: Mitral annulus function, particularly when work and power are calculated, appear to be an important determinant of left ventricular filling. The underlying mechanics may also explain the variations in transmitral and pulmonary venous Doppler patterns seen in diseased states. PMID- 11433825 TI - Echocardiography and exercise electrocardiography in the assessment of long-term outcome after percutaneous mitral valvuloplasty. AB - Before and immediate (invasive examination) and long-term follow-up (4.5 year; echocardiography, exercise electrocardiography) studies after percutaneous transvenous mitral valvuloplasty (PTMV; Inoue technique) were prospectively performed in 123 consecutive patients with pure or dominant mitral stenosis. These patients consisted of two groups: Group A with suitable valve morphology (n = 108, mean age 49.6 years, echocardiographic score 8.2 +/- 1.5), and Group B with less favourable valve morphology (n = 15, mean age 59.3 years, echocardiographic score 11.3 +/- 2.3) rejected for surgery because of high surgical risk. PTMV is a safe and effective nonsurgical method of treatment for symptomatic mitral stenosis and has long-lasting results in patients with pliable mitral valve as well as in patients with less suitable valve morphology. Echocardiography (Doppler approach) is the most efficient noninvasive method for assessment of PTMV effect. Exercise electrocardiography test is a valuable complementary method for objective measurement of patient working capacity and to confirm the success of PTMV. PMID- 11433826 TI - No reflow phenomenon in coronary heart disease. AB - Prolonged myocardial ischemia often damages myocardium profoundly, and flow to the previously ischemic myocardium is significantly reduced, which is called the "no reflow" or "low reflow" phenomenon. Myocardial contrast echocardiography (MCE) shows that a not insignificant number of patients with acute myocardial infarction have a substantial no reflow phenomenon. Our studies have demonstrated that patients with MCE-no reflow have larger myocardial infarctions, poorer functional outcomes, more frequent complications, and left ventricular remodeling and higher mortality compared with those with MCE-reflow. MCE-no reflow is also the main cause of slow flow after coronary intervention for acute myocardial infarction. We can now improve microvascular function with adjunctive pharmacological intervention. Intracoronary verapamil and intravenous infusion of nicorandil in conjunction with reperfusion therapy augment myocardial perfusion and are associated with better functional and clinical outcomes compared with coronary reperfusion alone. Study of myocardial perfusion with MCE provides useful information for predicting the success of reperfusion therapy at the microcirculation level and the impact of pharmacological intervention on acute myocardial infarction. PMID- 11433827 TI - Noninvasive estimation of coronary flow reserve by transthoracic Doppler echocardiography with a high-frequency transducer. AB - Recent technological advances in transthoracic Doppler echocardiography (TTDE) provide visualization of the coronary blood flow signal and noninvasive analysis of coronary flow reserve with a high frequency transducer. Coronary flow velocity and coronary flow reserve measured by TTDE were compared with those measured by a Doppler guide wire. Values of mean coronary flow velocity and coronary flow reserve measured by TTDE were closely correlated with values measured by the invasive Doppler guide wire (r = 0.97, y = 0.94x + 0.40; r = 0.94, y = 0.95x + 0.21, respectively). This noninvasive technique was applied to patients who underwent coronary angiography to evaluate coronary artery disease. There was a significant difference in coronary flow reserve measured by TTDE between patients with and without significant left anterior descending coronary artery stenosis (% diameter stenosis > 70%) (1.5 +/- 0.2 vs 2.6 +/- 0.4, p < 0.001). Coronary flow reserve < 2.0 obtained by TTDE had a high sensitivity (92%) and a high specificity (86%) for the presence of significant left anterior descending coronary artery stenosis. TTDE with a high-frequency transducer can be applied to noninvasive estimation of coronary flow reserve in the clinical setting, and is useful in noninvasive diagnosis of significant coronary stenosis in patients with coronary artery disease. PMID- 11433828 TI - Noninvasive coronary imaging. AB - Noninvasive imaging of the coronary arteries is one of the most important but challenging goals in medical imaging. Recent methods being developed for visualization of the coronary arteries include transthoracic and transesophageal echocardiography, electron-beam computed tomography, and magnetic resonance imaging. Transesophageal echocardiography is a feasible noninvasive technique for imaging the left main coronary artery and detecting hemodynamically significant luminal obstruction. Recent technological advances in transthoracic Doppler echocardiography provide measurement of coronary flow velocity in the distal portion of left anterior descending artery, which is useful for the assessment of coronary flow reserve, and therefore coronary stenosis and restenosis after coronary intervention in left anterior descending artery lesions. Electron-beam computed tomography is a cross-sectional imaging technique with high spatial and temporal resolution. It is possible for image acquisition to be triggered by the patient's electrocardiogram and this technique is well suited to cardiac imaging. Electron-beam computed tomography might be useful to detect or rule out high grade coronary artery stenoses and occlusions, when image quality is adequate. Recent improvements and progress in coronary magnetic resonance angiography stem from the use of respiratory motion compensation and fast imaging methods. The most commonly used breath-hold method was developed with a fast segmented two dimensional sequence on thin sections oriented to capture the coronary vessel inplane. Sensitivity and specificity of coronary magnetic resonance angiography in detecting significant coronary artery disease are 90% and 92% using this method. PMID- 11433829 TI - Clinical roles of perfusion and metabolic imaging. AB - Noninvasive assessment of regional myocardial perfusion is clinically important for early and accurate diagnosis of coronary artery disease. In addition, persistent metabolic alterations are often seen in post ischemic dysfunction after recovery of blood flow. Thus, prior ischemic insult may be identified as areas of altered metabolism despite normal perfusion (so-called ischemic memory imaging). Radionuclide imaging has great advantages over other imaging techniques based on the variety of radiopharmaceutical agents to probe regional cellular functions and biochemistry in vivo. Technetium-99m perfusion imaging agents provide excellent myocardial perfusion images which may enhance diagnostic accuracy in the study of coronary artery disease. In addition, greater photon flux from the tracer permits simultaneous assessment of regional perfusion and function with electrocardiogram-gated acquisition. Positron emission tomography (PET) enables metabolic assessment in vivo. Preserved fluorodeoxyglucose (FDG) uptake indicates ischemic but viable myocardium which is likely to improve regional dysfunction after revascularization. In addition, FDG-PET seems to be valuable for selecting a high risk subgroup. Recently, iodine-123 15-(p iodophenyl)-3R, S-methyl pentadecanoic acid (BMIPP), a branched fatty acid analog, has become clinically available in Japan. Less uptake of BMIPP than thallium is often observed in the ischemic myocardium. This perfusion metabolic mismatch, which is also observed by FDG-PET, is identified with stunned or hibernating myocardium with regional dysfunction. Both are likely to recover afterwards. Severe ischemia is identified as reduced BMIPP uptake at rest, suggesting use as an ischemic memory imaging. These new techniques will provide insights into new pathological states in ischemic heart disease and help to select the optimal treatment for the patients. PMID- 11433830 TI - Noninvasive investigations of systolic and diastolic heart function in arterial hypertension: from calibrated and differentiated apexcardiography to echocardiography. AB - In the early stages of arterial hypertension, increased contractility, demonstrated by calibrated and differentiated apexcardiography is present. Already in this early phase of the disease the criteria of diastolic function, as during isovolumetric period (calibrated and differentiated apexcardiography, M mode echocardiography) and during transmitral flow, are abnormal. These disturbances occur a long time before a reduction of systolic function can be shown. Diastolic dysfunction is more frequent in hypertensives with left ventricular hypertrophy. With noninvasive methods it is possible to show that a primary neurohumoral overstimulation--enhanced contractility, normal systolic function--followed by cardiac remodelling and apoptosis with likewise early but progressive morphological disorders--diastolic dysfunction--are the main pathophysiological processes in hypertensive heart and vessel disease. They are decisive for the prognosis and represent the basis for rational organoprotective and also cardioreparative medical treatment. PMID- 11433831 TI - Noninvasive/invasive correlates of exaggerated ventricular interdependence in cardiac tamponade. AB - Ventricular interdependence is that property of the normal heart such that distension of one ventricle alters the distensibility and filling pressure of the other. This phenomenon coupled with reciprocal changes in right and left heart venous return during normal quiet respiration results in minor decreases in left ventricular stroke volume, systolic blood pressure, pulse pressure, total electromechanical systole (Q-A2), left ventricular ejection time and mitral e wave velocity during inspiration and minor increases in these parameters during expiration. Opposite changes in these parameters occur in the right heart with increases occurring during inspiration and decreases during expiration. Exaggerated ventricular interdependence occurs in cardiac tamponade when the pericardial constraint limits the total contents in the pericardial sac. This, together with the decreased effective left ventricular filling pressure which occurs during inspiration, is responsible for the exaggerated decrease in stroke volume, blood pressure, pulse pressure, left ventricular ejection time and mitral e-wave velocity in this condition. These observations, together with the echocardiographic findings of right atrial collapse, right ventricular collapse, and inferior vena cava plethora constitute the noninvasive diagnosis of pericardial tamponade. The utility of these noninvasive tests in detecting both the presence and degree of increased pericardial pressure was evaluated in 33 invasively studied patients with pericardial effusion. In Group 1 (n = 13) intrapericardial pressure was elevated but less than both right atrial pressure and pulmonary wedge pressure, in Group 2 (n = 10) intrapericardial pressure equaled right atrial pressure but was less than pulmonary wedge pressure, and in Group 3 (n = 10) intrapericardial pressure equaled right atrial pressure and pulmonary wedge pressure. From these data it is concluded that right atrial and right ventricular collapse are highly sensitive techniques for predicting increased intrapericardial pressure in all three groups, but fail to predict the level of intrapericardial pressure and the severity of hemodynamic compromise. However, the absence of inferior vena cava plethora helped separate Group 1 patients from Groups 2 and 3 patients, thereby aiding in distinguishing a group of patients with severe hemodynamic derangement requiring urgent intervention. Exaggerated reciprocal changes in mitral and tricuspid e-wave velocity is a very sensitive finding for increased intrapericardial pressure. However, its presence correlates poorly with the severity of tamponade. Acute pericardial tamponade is a clinical diagnosis determined by the integration of the history, physical exam and appropriate noninvasive physiologic and imaging techniques. PMID- 11433832 TI - Effects of centrally-acting glutamatergic modulators on cardiovascular responses to stress in humans. AB - Sympathetic overactivity is a common feature of certain cardiovascular diseases. An acute activation of the sympathetic nervous system can provoke angina pectoris attacks through the increase of myocardial oxygen demand, frequently associated to coronary arterial constriction. It can also promote cardiac arrhythmias leading, in some cases, to cardiac sudden death. The aim of the present study was to evaluate the cardiovascular effects of a single oral dose of baclofen or ifenprodil (two drugs modulating central glutamatergic relays) at rest and during three laboratory stressors (a cold pressor test, a mental arithmetic stress test and an exercise test on a cycloergometer), in human healthy volunteers. Ifenprodil increased resting heart rate and did not reduce the cardiovascular response to any test. In contrast, baclofen reduced the tachycardic response to mental stress test and so limited the increase of myocardial oxygen demand during the test. Nevertheless, this drug was not able to affect the cardiovascular response to exercise. Finally, we have shown in this study that baclofen exhibits a profile of a central sympathomodulator without cardiodepression. Its activity towards mental stress induced cardiovascular responses leads us to proposing this compound for testing after a chronic treatment, in patients with silent myocardial ischemia and mental stress test induced ischemia. PMID- 11433833 TI - Short-term heart rate variability in young athletes. AB - OBJECTIVES: The purpose of this study was to determine whether the type of training in a young population differentiates heart rate variability parameters between athlete groups and sedentary subjects. METHODS: The effect of different types of physical training on heart rate variability was evaluated in 10 aerobic trained athletes, in 7 anaerobic trained athletes, in 7 rugby players (mixed type training) and in 10 sedentary control subjects. All groups were age matched (18 34 years). Electrocardiogram tracings were recorded digitally in the supine position and standing position. Measures of heart rate variability were obtained, from both time- and frequency analysis of 10 min resting heart rate. After these tests, blood pressure was measured using an automatic inflation cuff. RESULTS: Resting heart rate was lower in aerobic and mixed type athletes compared to controls. Only aerobic athletes had evidence of increased vagal activity in the time domain compared with control subjects (increased SDNN supine, increased rMSSD supine and standing and pNN50 standing). In the frequency domain, aerobic athletes presented with both higher low-frequency and high-frequency power in the standing position and low-frequency power in the supine position compared to controls. CONCLUSIONS: It can be concluded that heart rate variability is affected by chronic exercise, especially in endurance trained athletes. This infers that especially aerobic exercising can have beneficial effects on the cardiovascular risk profile. PMID- 11433834 TI - Noninvasive evaluation of endothelial function. AB - Endothelial dysfunction is generally believed to be the inciting event in atherosclerosis, and is probably important in ischemic manifestations as well. The release of endothelium-derived vasoactive substances is not only triggered by acetylcholine, but also controlled by a host of neuromediators and by shear forces exerted by the blood flowing through the blood vessel. However, this balance is altered in disease states such as atherosclerosis, diabetes, chronic heart failure, coronary artery disease, or hypertension. The most important mechanism in the decrease in endothelium-dependent relaxation appears to be a reduced release of nitric oxide. In healthy people, the predominant effect of stimulation of the endothelium is vasodilation. It is tempting to hypothesize that endothelial dysfunction is one of the initial steps involved in the development of atherosclerosis, but also in peripheral artery atherosclerosis. Impairment of endothelium-dependent vasodilation in the coronary arteries has been demonstrated not only in patients with documented atherosclerosis and/or established cardiovascular risk factors. Noninvasive evaluation of brachial artery vasoactivity using high resolution B-mode ultrasound is currently being established to evaluate endothelial function. We studied the endothelial function in 50 normal volunteers, 28 hypertensive subjects, and 31 hypercholesterolemia subjects. The diameter of the target artery was measured from two-dimensional ultrasound images with 10 MHz linear transducer. The results suggested that antihypertensive therapy with a certain calcium antagonist did not have a favorable effect on endothelial function and after cessation of cholesterol lowering therapy, the endothelial dysfunction developed again. The endothelial function can now be readily measured in humans and is very useful research tool to assess the effect of risk factors and their treatment on vascular function. Endothelial function testing will assume a prominent role in the evaluation and treatment of patients at risk of developing coronary atherosclerosis and its sequelae. PMID- 11433835 TI - Real-time, volumetric echocardiography: usefulness of volumetric scanning for the assessment of cardiac volume and function. AB - BACKGROUND: A novel imaging system has been introduced which uses a dedicated two dimensional echo probe for rapid beam forming to scan a pyramidal volume in real time. Real-time volumetric echocardiography has the potential to determine accurate cardiovascular anatomy, volume and function in the beating heart without reconstructions. The results of animal and human studies using volumetric echocardiography are evaluated for the potential for clinical applications. IMAGING METHODOLOGY: A new type of ultrasound imaging, high-speed volumetric scanning based on phased array principles permits real-time three-dimensional, volumetric echocardiography (real-time 3-DE). The system requires no off-line reconstruction techniques, thus enabling dynamic three-dimensional visualization and quantification of the heart in real time using a transthoracic approach. Real time 3-DE uses a 2-D matrix phased array transducer. Image formation employs 16:1 parallel processing to scan a pyramidal volume composed of multiple steering directions in the azimuth dimension and in the elevation dimension. The finished transducer is mounted in a hand-held case with a circular aperture of 16 mm diameter. The array consists of approximately 1,600 elements, operating at 2.5 MHz. Real-time 3-DE permits simultaneous, multiple plane display of two sector arcs (B-scans) and C-scan (parallel to the transducer face or inclined) on a single monitor, conveying the three-dimensional nature of the ultrasound data. This system also allows these planes to be angled for extra diagnostic flexibility. The motion of all the structures during the cardiac cycle can be evaluated in dynamic mode. METHODS: Real-time 3-DE was assessed for accuracy of volume measurement by measuring the volume of balloons of different size and shape, and the hearts of 15 closed chest dogs with myocardial contrast enhancement, and compared to the volumes measured by left ventricular angiography in the dogs. Real-time 3-DE was used to evaluate the endocardial border determination of the entire left ventricle by injecting contrast agent in 12 patients. The endocardial border determination of each segment was scored, and the endocardial border score index calculated. Both real-time 3-D images and cine magnetic resonance imaging (MRI) were performed in 16 patients to assess the accuracy of volume measurement of the left ventricle in humans. The endocardial border of the left ventricle was manually traced, and the volumes calculated by Simpson's rule. RESULTS: The volumes measured by real-time 3-DE correlated well with the true volumes for different sizes of balloon and for asymmetric balloons. The end-diastolic volume and end-systolic volume linear correlation of real-time 3-DE versus angiography measurements using manual tracing in vivo also gave a good correlation (r = 0.97, p < 0.001; r = 0.92, p < 0.01). Fifty-eight of 192 segments were rated as good at baseline and 143 rated as good after Levovist injection. Endocardial border determination was improved by Levovist injection in 100 of 137 segments (74.6%). The endocardial border score index was significantly higher after Levovist administration than at baseline (p < 0.003). The end diastolic volume and end-systolic volume of the left ventricle measured by real time 3-DE in humans correlated well with those measured by MRI (end-diastolic volume: r = 0.97, p < 0.001; end-systolic volume: r = 0.96, p < 0.001). CONCLUSIONS: Transthoracic real-time, volumetric echocardiography opens a new and exciting field of echocardiography. The results of these studies demonstrate that this system can accurately measure the ventricular volume and function without use of geometric assumptions. This volumetric mode or V-mode scanning is a new imaging modality that provides a practical methodology to investigate important clinical and research questions. PMID- 11433836 TI - Congenital anomaly: middle ear malformation. PMID- 11433837 TI - Endoscopic view of a mucocele in an infraorbital ethmoid cell (Haller cell). PMID- 11433838 TI - Amyloidosis of the larynx. PMID- 11433839 TI - ENG in a 70-year-old woman with dizziness and inability to look down. PMID- 11433840 TI - Mandibular mass, osteosarcoma. PMID- 11433841 TI - Bilateral peritonsillar abscess: case report and presentation of its clinical appearance. AB - Although there is no consensus on its incidence, bilateral peritonsillar abscess is an unusual variant of an otherwise relatively common otolaryngologic disease. A bilateral peritonsillar abscess can be differentiated from other oropharyngeal pathology with a detailed physical examination and complementary imaging. Its diagnosis should always be considered in patients who have signs and symptoms that are suggestive of peritonsillar abscess but whose intraoral examination yields atypical findings, as well as in patients with marked distress or trismus. This article describes the case of a young man who came to the emergency room with bilateral peritonsillar abscess. The author believes that this report contains the only published photograph of the intraoral appearance of this condition. PMID- 11433842 TI - A new case of a branchial cyst of the parapharyngeal space. AB - Branchial cysts in the parapharyngeal space are rare. Until now, only 23 such cases have been reported in the literature. In this article, we report a new case in a 65-year-old man. Information gained from the clinical examination, fiberoscopy, and computed tomography revealed that the cyst was obstructing the oropharynx and filling the parapharyngeal and retropharyngeal spaces on the right. The mass was excised via a transcervical approach without any complications. We also review the literature on this condition. PMID- 11433843 TI - Endoscopic physiologic approach to allergy-associated chronic rhinosinusitis: a preliminary study. AB - Patients with allergy-associated chronic rhinosinusitis respond poorly to functional endoscopic sinus surgery (FESS), probably because of the altered nasosinus ventilatory physiology and the increased contamination of the nasosinus mucosa by the offending allergens. With this in mind, we describe the concept and technique of functional endoscopic nasosinus surgery (FENS) in such cases. The advantages of this technique are that it preserves the uncinate process, it limits ethmoidal exenteration via a transbullar approach, and it simultaneously corrects both septal and lateral wall pathologies. The aim of our randomized prospective study was to subjectively and objectively compare the efficacy of FESS and FENS in 64 patients with allergy-associated chronic rhinosinusitis. Subjective assessment was ascertained by visual analog scores, and objective assessment was made by nasal endoscopy. In this preliminary study, we found that FENS was superior to FESS in treating chronic sinusitis associated with nasal allergy. PMID- 11433844 TI - Vestibular and hearing manifestations of phenytoin toxicity: a retrospective series. AB - We undertook a study to identify the vestibular and hearing abnormalities associated with phenytoin toxicity. Since October 1977, 23 patients with phenytoin toxicity have been referred to the Department of Otorhinolaryngology by the Department of Neurology for electrooculographic (EOG) evaluation. (There have been no cases of serious phenytoin toxicity seen in our department since August 1989.) In addition to EOG, all patients had undergone otoscopic, audiometric, and neurologic examinations. We found that the most common pathologic findings detected by EOG were abnormalities in pursuit tracking. The most common signs found on clinical exploration were cerebellar dysfunction and facial palsy. EOG findings in patients with phenytoin toxicity are compatible with cerebellar disease. PMID- 11433845 TI - Nasal glioma: a case report. AB - Nasal gliomas are rare, benign, congenital tumors that are thought to be the result of an abnormality in embryonic development. Three types of clinical presentations have been recognized: extranasal (60%), intranasal (30%), and combined (10%). Clinically, these masses are firm, noncompressible, nonpulsatile, gray or purple lesions that obstruct the nasal cavity intranasally and cause deformity extranasally. Histologically, these tumors are made up of astrocytic neuroglial cells interlaced with fibrous and vascular connective tissue that is covered with skin or nasal respiratory mucosa. The preferred treatment is excision via a frontal craniotomy approach for those patients whose gliomas have an intracranial connection, for those patients who have cerebrospinal fluid rhinorrhea, and for those who have recurrent episodes of meningitis. Otherwise, a transfacial lateral rhinotomy is advised. In this article, we describe a case of a nasal glioma that was removed via a lateral rhinotomy. PMID- 11433846 TI - Forever vigilant. PMID- 11433847 TI - Sex vs. gender. PMID- 11433848 TI - Implants and endodontics. I. PMID- 11433849 TI - Implants and endodontics. II. PMID- 11433850 TI - Allied health professionals. PMID- 11433851 TI - Solving the shortage. PMID- 11433852 TI - Aphthous ulcers revisited. PMID- 11433853 TI - Comparing efficacy and safety of four intravenous sedation regimens in dental outpatients. AB - BACKGROUND: Management of patients' fear and anxiety during dental treatment is a primary concern of dental practitioners. Pharmacological strategies used in outpatient dental settings must be both safe and effective. Regimens of intravenously administered sedative drugs were evaluated in a collaborative, multicenter study of outpatients undergoing removal of impacted third molars. METHODS: A total of 997 patients randomly received one of five treatments: placebo; midazolam administered to a clinical endpoint of conscious sedation (mean dose, 8.6 milligrams); midazolam plus additional midazolam as needed during the procedure (mean total dose, 12.2 mg); fentanyl (1.4 micrograms/kilogram) plus midazolam to achieve the same endpoint of conscious sedation (mean dose, 5.7 mg); or fentanyl (1.4 (micrograms/kg), midazolam (mean dose, 5.8 mg) and methohexital as needed during the procedure (mean dose, 61.0 mg). RESULTS: Each drug regimen reduced anxiety during surgery in comparison with placebo, with the combination of midazolam, fentanyl and methohexital resulting in significantly less anxiety in comparison with the other treatment groups. Pain reports by patients during surgery also were reduced significantly by the combination of fentanyl, midazolam and methohexital. Patients' global evaluations of the efficacy of sedation ranked midazolam with supplemental midazolam and the combination of fentanyl, midazolam and methohexital as significantly more efficacious than the other two drug regiments. The authors noted transient respiratory depression in patients in the two opioid-treated groups, but no other physiological changes were detected. CONCLUSIONS: These data provide evidence that the drugs and doses evaluated resulted in therapeutic benefit to dental outpatients, with minimal incidence of potentially serious adverse effects. CLINICAL IMPLICATIONS: The results of this large-scale study provide assurance to both the public and the dental profession of the safety of parenteral sedation with these drugs and combinations of these drugs when titrated slowly in the recommended doses by appropriately trained dentists. PMID- 11433854 TI - Diagnosing TMJ internal derangement and osteoarthritis with magnetic resonance imaging. AB - BACKGROUND: The authors conducted a study to investigate the relationship between the presence of temporomandibular joint, or TMJ, pain and magnetic resonance imaging, or MRI, findings of internal derangement, or ID, and arthritis, or OA. METHODS: The authors studied 131 consecutively seen TMJ pain patients. Their criteria for including a patient in the study were report of unilateral orofacial pain referred to TMJ and the presence of unilateral TMJ pain during palpation, function, and unassisted or assisted mandibular opening. The authors took bilateral sagittal and coronal MRIs to establish the presence or absence of TMJ ID, OA or both. RESULTS: When the authors compared TMJ side-related data from all subjects, they found a significant relationship between the clinical findings of TMJ pain and the MRI diagnoses of TMJ ID (P = .000) and TMJ OA (P = .013). They also found a significant relationship between the MRI diagnosis of TMJ ID type and that of TMJ OA (P = .000). The authors used the kappa statistical test and found poor diagnostic agreement between the presence of TMJ pain and the MRI diagnosis of TMJ ID (kappa = 0.21), TMJ OA (kappa = 0.15), and TMJ ID and OA combined (kappa = 0.18). CONCLUSIONS: The study's findings suggest that while clinical pain is related to TMJ-related MRI findings, the presence of clinical pain is not a reliable predictor of TMJ ID, OA or both. CLINICAL IMPLICATIONS: Using MRI to supplement clinical findings of TMJ pain appears warranted and necessary to establish the presence or absence of TMJ ID. PMID- 11433855 TI - A clinical evaluation of air-abrasion treatment of questionable carious lesions. A 12-month report. AB - BACKGROUND: The efficacy of treating questionable incipient lesions early with air abrasion, a modality used by many practitioners, has not been adequately demonstrated. METHODS: The authors enrolled 223 teeth, each with a questionable incipient pit-and-fissure carious lesion, from 93 dental patients in a projected five-year randomized clinical trial. Caries was defined as softness, decalcification or cavitation at the base of a pit or fissure or radiographic evidence of caries. Each tooth was randomly assigned to either a treatment group (n = 113 teeth) or a control group (n = 110 teeth) (which was observed but left untreated until the definition of caries was met). Each tooth in the treatment group was air-abraded and restored with a flowable resin-based composite. The authors re-examined teeth in both groups every six months; they evaluated the restorations using a modified set of Ryge criteria and inspected teeth for caries using radiographs, mirrors and standardized explorers. RESULTS: Of the 113 teeth with questionable incipient carious lesions air-abraded in the treatment group, 50 had caries extending into dentin. After 12 months of clinical service, there were three sealants that exhibited a partial loss of sealant which did not require any re-treatment. Two restorations with penetrating staining were re treated. In the control group at the end of 12 months, only nine of the 86 recalled teeth were diagnosed with pit-and-fissure caries and were treated with air abrasion and restored with flowable resin-based composite. There was no statistically significant difference between the volume of the treatment and control preparations. CONCLUSION: After 12 months of clinical service, two preventive resin-based composite restorations in the treatment group required re treatment. Fewer teeth than expected in the control group were diagnosed as having caries and were treated. CLINICAL IMPLICATIONS: The merit of treating questionable incipient pit-and-fissure carious lesions early with air abrasion has not been demonstrated after 12 months in this clinical study. PMID- 11433856 TI - Occlusal stabilization appliances. Evidence of their efficacy. AB - BACKGROUND: There is substantial controversy regarding the value of occlusal appliances for managing temporomandibular joint disorders. This article specifically assesses whether the evidence is sufficient to judge occlusal appliances as being efficacious for the management of localized masticatory myalgia, arthralgia or both. A major confounder is that few studies have measured or evaluated whether subjects had strong, ongoing parafunctional activity (such as clenching or grinding) and whether appliances influenced this behavior. LITERATURE REVIEWED: The authors evaluated four placebo-controlled studies, several randomized wait-list controlled studies and several random-assignment treatment-comparison studies. Data from the wait-list condition studies vs. those from the occlusal appliance condition studies consistently suggested that the latter treatment's effect on patient symptom level is far more than that of no treatment on a wait-list group's condition. In contrast, the studies on placebo controlled vs. occlusal appliance studies yielded a mix of data: two showed a positive benefit of occlusal vs. nonoccluding appliances, and two showed a null effect or no difference. CONCLUSIONS: Considering all of the available data (pro and con), the authors conclude that the use of occlusal appliances in managing localized masticatory myalgia, arthralgia or both is sufficiently supported by evidence in the literature. CLINICAL IMPLICATIONS: The mechanism of action by which occlusal appliances affect localized myalgia and arthralgia probably is behavioral modification of jaw clenching. However, if the behavior continues unabated, even the best splint will not work. PMID- 11433857 TI - Using core markers to enhance visualization of the core material/tooth interface. PMID- 11433858 TI - A fast and easy method of fabricating I-bar clasps in the dental office. PMID- 11433859 TI - Is it possible to control the directional shrinkage of resin-based composites? PMID- 11433860 TI - Stress-related suicide by dentists and other health care workers. Fact or folklore? AB - BACKGROUND: The media repeatedly portrays dentists and other health professionals as being at risk of committing suicide. While this message often is accepted without question, there are little reliable data available that verifies this alleged risk. The relationship between professional stress and suicide, if any, has not been substantiated or quantified. TYPES OF STUDIES REVIEWED: The author evaluated the contemporary literature on stress and suicide in health professionals in an effort to verify or refute the widely held belief that dentists and other health care professionals are at higher risk of committing stress-related suicide. The author also surveyed dental schools to determine what efforts were being made to provide students with stress-management skills. RESULTS: The author found that there is little valid evidence that dentists are more prone to suicide than the general population, although some related data suggest that female dentists may be more vulnerable. Large-scale studies are needed before firmer conclusions can be reached. The author's survey shows that dental students generally receive some education on stress management, but many dental hygiene and graduate students do not. The author makes several recommendations for future research. CLINICAL IMPLICATIONS: Although some dentists leave the profession by way of suicide or career change at a time when their careers should be the most rewarding, available data on stress and its impact on suicide incidence are inconclusive and flawed. The profession needs to identify the causes of stress-related suicides and provide assistance to those people who are affected by stress. PMID- 11433861 TI - How can dentists and parents communicate effectively about children's dental health needs? PMID- 11433862 TI - Where will the genome lead us? Dentistry in the 21st century. AB - BACKGROUND: Recent announcements of the deciphering and analysis of the human genome signal the inception of a new era of gene-based medicine. During the 21st century, patient treatment will be transformed and dentistry will be affected profoundly. METHODS: The author explains the importance of the decoding of the genome and how--based on this now completely depicted molecular structure--genes build, maintain and control all the biological functions of humans and all other living organisms. The potential application of this knowledge to the practice of dentistry is addressed, as well as the ethical, legal and moral challenges to the profession engendered by this new technology. CONCLUSION: During the next several decades, many of the current materials and methods will be abandoned in favor of emerging bioengineered technologies, genetically programmed for the prevention and treatment of oral disease as well as for the repair of damaged dental tissues. PRACTICE IMPLICATIONS: The development and implementation of these innovative dental therapies will require intensive education of current practitioners. Considerable restructuring of dental school curricula will need to take place, and the emergence of a new dental specialty is anticipated. PMID- 11433863 TI - Cast gold restorations. Has the esthetic dentistry pendulum swung too far? PMID- 11433864 TI - Illinois court finds a catch in restrictive covenants. PMID- 11433865 TI - Tough choices. PMID- 11433866 TI - The care crisis. PMID- 11433867 TI - Minnesota's nursing shortage. PMID- 11433868 TI - Focusing on the first critical years. PMID- 11433869 TI - Nursing. Something for everyone. PMID- 11433870 TI - Nurses without borders. PMID- 11433871 TI - Following the leaders. Minnesota's nurses continue a tradition of leadership in health care. PMID- 11433872 TI - Success is near on public health bills. PMID- 11433873 TI - Interdisciplinary collaboration in the 21st century. PMID- 11433874 TI - The Southeast Minnesota Fetal and Infant Mortality Reduction Project. AB - Using linked birth and death certificates, researchers collected information on the events surrounding and potentially contributing to all fetal and infant deaths in an 11-county southeast Minnesota area May 1, 1996, through May 31, 1998. Data were collected from medical record reviews and maternal interviews. Information was collated and presented to county review teams consisting of health, human services, and education personnel. These teams identified factors associated with potentially preventable deaths and gaps in services for women, infants, and families. A regional team then grouped and prioritized the factors. PMID- 11433875 TI - Listening to nurses. PMID- 11433876 TI - Bipolar illness and society. Introduction. PMID- 11433877 TI - Bipolar illness and the family. AB - The purpose of this article is to introduce family systems therapy and "family focused treatment" to clinicians working with patients who are diagnosed with bipolar illness. The characteristic traits of healthy family functioning are identified as a template for the clinician to work with the family toward management of bipolar illness. Nathan Ackerman's model of how families handle crises and Carter's and McGoldrick's work with family life cycles provide a guide to understanding family systems work. The focus is on the family system and not just the patient. The issues of bipolar illness are addressed in the family context. Based on the research of Goldstein and Miklowitz and others, treatment of bipolar illness is differentiated from other chronic mental illnesses. Family focused treatment is offered as a model to manage dynamics off bipolar illness. An essential component of therapy with the patient and the family is addressing grief. Interventions and treatment suggestions are offered. PMID- 11433878 TI - Bipolar disorder and violence. AB - Violent behavior presents many social, legal, and clinical problems. A number of models have been developed to explain violence, representing a variety academic disciplines and theoretical orientations. Unfortunately none of these approaches have led to a comprehensive understanding of violence and aggression. The issue of violence is particularly perplexing in connection with the role of psychiatric disorders as contributing factors. Several psychiatric conditions, including bipolar disorder, have been implicated with increased rates of violent behavior. This paper examines various models of violence, influences on violent behavior, and violence associated with psychiatric disorders. Particular attention is devoted to the connection between bipolar disorder and violence. PMID- 11433879 TI - Bipolar illness, creativity, and treatment. AB - There have been in recent years increasing claims in both popular and professional literature for a connection between bipolar illness and creativity. A review of studies supporting this claim reveals serious flaws in sampling, methodology, presentation of results, and conclusions. Although there is therefore no evidence for etiological or genetic linkages, it is still necessary to explain interrelationships in those creative persons suffering from the illness. Examples of the work in progress of artists with bipolar disorder, Jackson Pollock and Edvard Munch, illustrate the use of healthy and adaptive creative cognition--janusian and homospatial processes--in the former's breakthrough conception during an improvement phase in treatment leading to the development of the Abstract Expressionist Movement and in the latter's transformation of an hallucination into his famous artwork "The Scream." Treatment options that do not produce cognitive effects are important for creative persons with bipolar disorder. PMID- 11433880 TI - Lithium revisited: savings brought about by the use of lithium, 1970-1991. AB - BACKGROUND: Recent estimates of the cost of manic-depressive illness totaled roughly $45 billion in 1991. Using data from the Epidemiological Catchment Area (ECA) study, this study estimates the savings brought about by the use of lithium between 1970 and 1991. METHODS: Total savings are the difference between estimated actual costs and projected costs had lithium never been introduced. Actual yearly costs were interpolated from data for 1970 and 1991, and projected costs were obtained by adjusting 1970 costs with Consumer Price Index (CPI) and population inflaters. All costs for 1970 were obtained using methods almost identical to those used to calculate the 1991 costs of manic-depressive illness, presented in a previous publication. All savings are presented in 1991 dollars. RESULTS: Between 1970 and 1991, lithium saved over $170 billion, or roughly over $8 billion per year. Approximately $15 billion in direct costs, which included inpatient and outpatient care as well as research, was saved between 1970 and 1991. The savings are more dramatic for indirect costs, which include the lost productivity of wage-earners, homemakers, family caregivers, and individuals who are in institutions or who committed suicide; these totaled roughly $155 billion. CONCLUSIONS: Our results suggest that, although manic-depressive illness is still costly, lithium has been tremendously successful in treating the illness, and has provided enormous financial savings in the process. PMID- 11433881 TI - Patients' perceptions of their bipolar illness in a public hospital setting. AB - This preliminary study explored the perceptions of illness of people in a public hospital setting for treatment of bipolar disorder. Fifteen inpatients (5 African Americans, 5 Hispanics, and 5 European Americans) participated in audio-recorded, structured interviews. The interviews were designed to identify recurring themes and patterns in their perceptions of bipolar disorder and to assess their cognitive representations of the illness. The interviews were transcribed verbatim, and content analysis procedures were used to identify the illness perceptions and to compare them based upon ethnic orientation. Results indicated that most of the participants' group responses were more alike than they were different. However, discrete themes emerged among the individuals as their stories unfolded. These themes, and the perceived benefit of the interview itself, provide a beginning foundation for further research in the area, and have clinical relevance for health care providers working with this population. PMID- 11433882 TI - Substance use and other psychiatric disorders in impaired practitioners. AB - This paper describes the nature and comorbidity of substance use and other psychiatric disorders in 157 impaired practitioners notified to the Medical Practitioners Board of Victoria from 1983 to 1997. Diagnoses were reduced to three diagnostic categories, viz, substance use disorder (n = 62), other psychiatric disorder (n = 62), and combined substance use and other psychiatric disorder (n = 33). Sixty nine percent of those with other psychiatric disorders suffered either bipolar disorder (n = 27), schizophrenia (n = 22), or depression (n = 17). Psychiatric comorbidity in drug-related diagnoses was 26% and in alcohol-related diagnoses 64%. Sixty five percent of substance use disorders were notified more than 12 months after onset of impairment, and 49% of all practitioners (71% with substance use disorder) did not receive treatment prior to notification. This combined with a high relapse rate (41%) makes some impaired practitioners a risk to their patients. PMID- 11433883 TI - Clonidine abuse among opiate addicts. AB - OBJECTIVE: To determine if clonidine abuse among non-pregnant opiate dependent individuals is common and the reason for such use. METHOD: Self report of personal use or knowledge of others' use of this drug. RESULTS: Fourteen of fifteen treatment seeking individuals with opiate dependence knew of clonidine abuse. Ten had used it personally to decrease the amount of heroin necessary to achieve a desired effect and to prolong the length of the opiate's action. CONCLUSION: Clonidine abuse among opiate addicts may be more common than previous studies have suggested. Opiate addicts should be screened for abuse of this substance, especially in view of its widespread use for the purpose of opiate withdrawal. Further, such patients should be warned of potential health hazards attendant upon clonidine use and abrupt cessation. PMID- 11433884 TI - Use of propranolol in heart failure patients: safety, tolerability, and effects on left ventricular function. AB - INTRODUCTION AND OBJECTIVES: Beta-blockers have been shown to improve prognosis in patients with heart failure (HF). Propranolol, which is a low-cost drug, has not been fully studied in this setting. We sought to determine the safety, tolerability and effects on left ventricular function observed with the use of propranolol in HF patients, in functional class II-IV of the New York Heart Association. POPULATION AND METHODS: Prospective study in which 20 outpatients (10 male, mean age 56 +/- 12 years, ranging from 20 to 70) were included. Mean left ventricular ejection fraction (EF) was 28%. Safety, tolerability and effects on electrocardiographic and echocardiographic variables were analyzed. Patients were evaluated in three steps: a) Step I--optimization on conventional drugs and assessment of baseline parameters; b) Step II--start of propranolol (10-20 mg/day), increasing the dose weekly to achieve a heart rate of 60 bpm, or a maximum daily dose of 120 mg; c) Step III--reappraisal of the parameters analyzed in step I, after 3 months of propranolol treatment. RESULTS: On average, after treatment with propranolol, EF increased by 52% (p = 0.0003), E wave deceleration time was prolonged by 62% (p = 0.001) and effective ventricular filling time increased by 38.5% (p = 0.0005). Two patients developed mild congestion which was controlled by increasing diuretic doses, with no need to interrupt the protocol. Four patients had bradycardia-related symptoms, controlled by reducing digoxin doses. Nine subjects developed hyperkalemia, reversed by interrupting or reducing spironolactone. CONCLUSION: Propranolol was safe and well tolerated, and had beneficial effects on ventricular function in HF patients. Its impact on mortality requires further study. PMID- 11433885 TI - Beta-adrenergic blockers and heart failure. PMID- 11433886 TI - [Can blood homocysteine explain the family history of vascular diseases?]. AB - INTRODUCTION AND AIMS: Family history of vascular disease is an important risk factor for vascular disease, independent of conventional risk factors. Homocysteinemia, a newly defined risk factor, is caused by genetics, such as cystathionine beta synthase deficiencies, and metabolic deficiencies. With the present work we intend to study the influence of family history of vascular disease in homocysteinemia. METHODS: We studied 204 normal persons (153 males), average age 38.7 +/- 10.9 years, in terms of family history of vascular disease (death due to myocardial infarction or a stroke), conventional risk factors, routine laboratory tests, fasting homocysteinemia and after oral methionine loading (0.1 g/Kg body weight). We compared laboratory results, conventional risk factors and homocysteinemia levels in persons with and without a family history of vascular disease. We performed covariance analysis to evaluate, in a multivariate model, factors that were related to basal or after methionine loading homocysteinemia. RESULTS: 35% of persons presented a family history of vascular disease (FHVD). Persons with FHVD presented higher age (45.6 +/- 8.9 versus 35.0 +/- 10.1, p < 0.001), and higher prevalence of hypertension (p = 0.002), dyslipidemia (p = 0.001), obesity (p = 0.03), and physical inactivity (p = 0.03). They presented a tendency, without statistical significance, to have a higher prevalence of diabetes and of hyperhomocysteinemia, and to present higher levels of basal and afterload homocysteinemia. Performing covariance analysis, basal homocysteinemia did not present any relation to FHVD. After methionine load homocysteinemia was strongly influenced by basal homocysteinemia (p = 0.0000), and significantly related to FHVD (p = 0.039). CONCLUSIONS: Homocysteinemia cannot explain most of the risk of family history of vascular disease, not explained by conventional risk factors. The only significant relationship between homocysteinemia and FHVD was observed with afterload homocysteinemia in the multivariate model. FHVD is clearly related to conventional risk factors. PMID- 11433887 TI - [Prognostic indexes derived from ventricular morphology in congenital diaphragmatic hernia induced by nitrofen]. AB - BACKGROUND: Some indices derived from cardiac morphology have been proposed as prognostic factors in fetuses and newborns with congenital diaphragmatic hernia (CDH). These would be of interest in evaluating the risk of pulmonary hypoplasia and the need for surgery in the fetus. The aim of this study was to evaluate the heart-related indices in the nitrofen-induced CDH rat model. METHODS: At day 9 1/2 of gestation dated pregnant female Wistar rats received 100 mg of nitrofen. Fetuses were harvested at day 21 1/2 by laparotomy. The fetuses exposed to nitrofen without CDH were defined as the control group (n = 38) while the fetuses exposed to nitrofen with CDH were defined as the CDH group (n = 30). The hearts from both groups were weighed, processed for paraffin embedding and fragmented in serial 7 microns thick transverse sections. In the hearts without cardiac morphological defects the right and the left ventricular cavity volumes were estimated. These allowed calculation of cardioventricular and left-ventricular mass indices. RESULTS: In the fetuses without cardiac malformations, no significant difference was found regarding heart-to-body weight ratio, cardioventricular index or left ventricular mass between the study groups. CONCLUSION: In this model the presence of CDH was not associated with any alteration in cardiac dimension, in the absence of structural malformations. These results suggest that the indices derived from ventricular morphology should not be used to evaluate the prognosis in fetuses and newborns with CDH. PMID- 11433888 TI - Spontaneous closure of congestional coronary fistula. AB - Coronary fistula is a rare congenital anomaly. In the pediatric population they are usually asymptomatic and this entity is suspected from the presence of a continuous murmur. They usually have a favorable course, although rarely they can complicate or occlude spontaneously. Early treatment is generally advised. The authors describe a case of spontaneous closure of congenital right coronary fistula draining into the right ventricle in a twelve-year-old boy and they comment on the therapeutic management of this anomaly. PMID- 11433889 TI - [Diabetes mellitus and ischemic cardiopathy: potential effects of the use of sulfonylureas]. AB - Morbidity and mortality are increased in diabetic patients with ischemic heart disease. Sulfonylureas are the therapeutic group most frequently used and contributing to this. Patients usually treated with sulfonylureas have increased cardiovascular mortality. This is probably attributed to the action of these drugs on the ATP-sensitive potassium channels via blocking the ischemic preconditioning. This article puts forward evidence on sulfonylureas cellular and molecular pharmacodynamics and the real clinical incidence in the ATP-sensitive potassium channels inhibition in the diabetic patient's ischemic heart. PMID- 11433890 TI - [Clinical guidelines and the practice of scientific evidence-based cardiology. Part I]. AB - Scientific medical information presents special characteristics, particularly its huge quantity as well as the speed with which it evolves as a result of cardiological research. These facts create difficulties in transferring research results into practice, thereby delaying the benefits of medical advances to patients. One of the possible solutions to this problem is the development, dissemination and implementation of clinical practice guidelines, as a clinical decision support system as well as a quality assurance methodology. The process of developing clinical practice guidelines involves a needs assessment, individual formulations, implementation and evaluation, as well as periodic revisions. It constitutes a complex activity, requiring technical and scientific resources for a good design and an efficacious practical implementation. In this article--as well as the next--we will present and discuss the methodology of creation, structure, evaluation and implementation of clinical practice guidelines, as a clinical decision support system as well as a quality assurance methodology in cardiology. PMID- 11433891 TI - Percutaneous balloon angioplasty in a pediatric patient with superior vena cava obstruction after cardiac surgery. PMID- 11433892 TI - [National electrophysiology registries for the year 2000]. PMID- 11433893 TI - Surgical anterior ventricular endocardial restoration (SAVER) in the dilated remodeled ventricle after anterior myocardial infarction. PMID- 11433894 TI - Restorative proctocolectomy with ileal pouch anal anastomosis in obese patients. AB - BACKGROUND: Obesity is a relative contraindication to performing restorative proctocolectomy. The aim of this study was to assess the morbidity and functional results after restorative proctocolectomy in obese patients as compared to a matched cohort of non-obese patients. METHODS: 334 patients who had restorative proctocolectomy were reviewed; obesity was defined as a body mass index (BMI) greater than or equal to 30 kg/m2. 31 obese patients were matched to 31 non-obese patients for age, gender, steroid use, and diagnosis. Operative time, length of hospitalization, and both perioperative (< 6 weeks) and long-term morbidity (> 6 weeks), especially sepsis, were evaluated. RESULTS: The BMI was significantly higher in the obese group (33.7 vs 23.2) (p < 0.0001), and no difference was found between the obese and non-obese groups relative to the matched parameters of age, gender, steroid use and diagnosis. There was no difference in the rate of mucosectomy performed between the obese and non-obese patients (9.6% vs 3.2%, p = NS). 16% of the obese patients underwent one stage restorative proctocolectomies as compared to 10% in the non-obese group. Operative time was longer in the obese group (229 min vs 196 min; p = 0.02), but overall hospital length of stay was similar (9.7 days vs 7.7 days; p = 0.13). Perioperative morbidity was higher in obese patients (32% vs 9.6%, p = 0.058). However, there was no statistical significance in long-term morbidity (23% vs 32%, p = 0.57) at a mean follow-up of 51 months in the obese group and 53 months in the non-obese group. Obese patients had more stomal complications (10 vs 0%) and incisional hernias (13 vs 3%) (p = NS). Overall the pelvic sepsis-rate was significantly higher in the obese group (16 vs 0%; p < 0.05). 60% of the obese patients who developed pelvic sepsis had pouch-anal anastomosis performed without proximal fecal diversion. Mean bowel movements/24 hours, pad use, nocturnal evacuation, accidents/24 hours and incontinence scores were not statistically significant between the groups. CONCLUSION: Obese patients have a higher rate of pelvic sepsis and peri-operative morbidity when compared to a matched non-obese cohort of patients; however, the functional outcome of restorative proctocolectomy in obese patients is not significantly different than in non-obese patients. PMID- 11433895 TI - Leptin and HDL-cholesterol in non-diabetic normotensive subjects. AB - BACKGROUND: We investigated the relationships between body mass index (BMI), serum leptin and serum HDL-cholesterol. MATERIAL AND METHODS: A retrospective study was carried out in 80 patients who did not have type 2 diabetes mellitus and/or high blood pressure. RESULTS: Both serum leptin and HDL-cholesterol serum levels correlated with BMI (r = 0.616 and r = -0.269, respectively), but when the BMI values were kept constant no correlation was found between serum leptin and HDL-cholesterol both in simple and in multiple regression. CONCLUSION: The findings suggest that serum leptin concentration is completely independent of lipid metabolism. PMID- 11433896 TI - Are there predictive factors of severe liver fibrosis in morbidly obese patients with non-alcoholic steatohepatitis? AB - BACKGROUND: Non-alcoholic steatohepatitis (NASH) is a clinicopathological entity characterized by the presence of steatosis and lobular and/or portal inflammation with or without fibrosis. Patients with non-alcoholic fatty liver and fibrosis on liver biopsy have increased liver-related deaths. METHODS: 181 wedge liver biopsies, taken at the time of bariatric surgery from patients with a mean body mass index (BMI) of 47, were studied. In all cases, the liver biopsy was performed without knowledge of the patient's clinical and biochemical data, which were then examined with univariate and multivariate analysis. RESULTS: Diagnosis of NASH was established in 105 patients (91%); 74 patients (70%) showed mild steatosis, 20 (19%) had moderate inflammation and fibrosis, and 11 (10%) had steatosis with severe fibrosis. None of the liver biopsies showed cirrhosis. Age was the only independent predictor of moderate and severe fibrosis (p = 0.001). CONCLUSIONS: Since only age was a predictor of moderate or severe fibrosis, and no clinical or biochemical abnormalities detected slowly progressive hepatic fibrosis, liver biopsy is the only means of detecting progression to more advanced liver disease in a NASH patient. PMID- 11433897 TI - Six years experience with minilaparotomy silastic ring vertical gastroplasty. AB - BACKGROUND: In 1994, we modified the silastic ring vertical gastroplasty (SRVG) procedure to be able to perform it through a small incision. We expected this MiniSRVG to reduce postoperative pain and discomfort, shorten hospital stay and cost, and reduce scars and overall morbidities. METHODS: From October 1991 to December 1999, 893 patients were operated for morbid obesity. From October 1991 to December 1993, 111 patients underwent the classic Eckhout SRVG. From January 1994 to February 1999, 782 patients underwent the MiniSRVG, in which the dissection is limited to the lesser curvature of the stomach and is done partly blindly. Certain technical maneuvers were done to facilitate the procedure and to shorten the incision. RESULTS: Immediate postoperative complications and overall morbidities were similar in both series. Long-term follow-up showed no significant differences in weight and BMI loss. The small incision in the MiniSRVG, however, shortened the median operating time (32.1 vs 38.1 minutes) and reduced greatly the incision size (6.5 vs 18 cm), the postoperative pain (1.2 vs 2.6 days), the hospital stay (3.0 vs 5.1 days), the evisceration rate (0.1 vs 2.7%) and incisional hernia rate (5.4 vs 15.8%). The only side-effect was an increase in seroma formation (11.8 vs 4.50%). CONCLUSIONS: The MiniSRVG was as safe and efficient as the classic SRVG, obtaining the same BMI reduction and satisfaction. PMID- 11433898 TI - Weight loss following vertical banded gastroplasty: intermediate results of a prospective study. AB - BACKGROUND: Morbidly obese patients who undergo purely restrictive bariatric operations may fail to maintain satisfactory long-term results. In an attempt to achieve the best possible outcome after restrictive procedures, we have employed preoperative selection criteria and are following this selected patient group over time in order to evaluate long-term success. MATERIALS AND METHODS: From June 1994 through August 2000, 166 morbidly obese patients underwent various bariatric procedures at our institution. Of these patients, 35 underwent vertical banded gastroplasty (VBG) based on selection criteria, including degree of obesity and dietary habits and eating behavior. All patients were seen at 1, 3, 6, 9, and 12 months postoperatively and yearly thereafter. Average follow-up time now is 4.1 years (29-75 mos.), and follow-up is 100%. A multivitamin and mineral supplement is administered to all patients for at least 6 months. Radiology examination is performed in all patients on the 4th postoperative day and at each yearly visit, in order to check for staple-line disruption and stomal stenosis. RESULTS: Early postoperative morbidity was 5.7%. Late postoperative morbidity was 22.8%. A significant number of patients had some degree of stomal stenosis as shown by radiology examination, but to date there has been no need for surgical revision. There has been no early or late mortality. Weight loss results expressed as average percent excess weight loss (% EWL) were as follows: 61% (28 90) at 1 year, 61% (20-90) at 2 years, 57% (13-91) at 3 years, 56% (25-87) at 4 years and 37% (24-59) at 5 years following surgery. A significant number of patients with excellent weight loss had a high frequency of vomiting. Evaluation by BAROS showed that 25% of patients had an overall unsatisfactory outcome. Anemia and iron deficiency were found in 46% and 32% of VBG patients respectively. Recurrence of preexisting comorbidities was significant if lost weight was regained. CONCLUSIONS: In spite of preoperative selection of patients for VBG, a significant percentage of patients had poor overall results in terms of weight loss, quality of life, and resolution of preexisting comorbidities. For these reasons and based on the long-term results published by others, VBG is no longer our preferred surgical option in morbidly obese patients. PMID- 11433899 TI - Laparoscopic vertical banded gastroplasty: early experience. AB - BACKGROUND: The tremendous development of laparoscopic surgery in the last decade is being applied to bariatric surgery. Laparoscopic vertical banded gastroplasty (LVBG) is technically feasible by laparoscopy. METHODS: From August 1998 to August 1999, 13 patients underwent LVBG. The technical difficulties are discussed. RESULTS: Operating time ranged from 105 to 420 minutes. 11 patients have lost 45-55% of their original weight. Inserting an esophageal bougie no. 11 from the beginning of the operation and using the laparoscopic set-up described, made the procedure shorter, safer and easier for patients and surgeon. CONCLUSION: LVBG is technically feasible for the laparoscopic bariatric surgeon who is experienced in the handling and control of laparoscopic instruments and hand-eye coordination. The early results are satisfactory. Preoperative counseling is an integral step for the operation. PMID- 11433900 TI - The mini-gastric bypass: experience with the first 1,274 cases. AB - BACKGROUND: Results of the laparoscopic Mini-Gastric Bypass (MGB) are reported. METHODS: 1,274 MGB patients are continuously monitored as part of an online computer tracking data-base system. RESULTS: Mean preoperative weight (+/- Standard Deviation) was 132 +/- 21 kg, BMI 47 +/- 7. Mean excess weight loss was 51% at 6 months, 68% at 12 months and 77% at 2 years. The mean operating-time was 36.9 +/- 33.5 minutes. The shortest time was 19 minutes. Hospital stay was 1.5 +/ 1.6 days. The overall complication rate has been 5.2%. The overall rate of deep vein thrombosis and pulmonary embolism was 0.08% and 0.16% respectively. The leak rate was 1.6%. There was one hospital death, 0.08%. Associated medical illnesses were either completely reversed or markedly improved. CONCLUSIONS: The MGB is safe, results in major weight loss, has a short operating-time, and has a short hospital stay. The MGB appears to meet many of the criteria of an "ideal" weight loss operation. PMID- 11433901 TI - Helicobacter pylori infection in patients undergoing gastric bypass surgery for morbid obesity. AB - BACKGROUND: Studies suggest that the incidence of Helicobacter pylori infection in obese patients, including those undergoing gastric reduction surgery, may be increased. METHODS: We examined the histologic findings at the time of surgery in a series of patients who were undergoing Roux-en-Y gastric bypass (RYGBP) for morbid obesity and compared these results with patients in our institution undergoing endoscopy. RESULTS: Of 60 patients undergoing RYGBP, material for histologic examination was available in 56 cases, and in 40 cases gastric fundic mucosa from the anastomotic site was sampled at the time of surgery. Active chronic gastritis was present in 6 (15%), and chronic gastritis was present in 27 (68%). H. pylori was present in all 6 cases of active chronic gastritis and in 9 cases of chronic gastritis (total 38%). This incidence of H. pylori infection was higher than that found in the series of gastric biopsies (107/500, 21%, p = 0.03) and fundic biopsies (10/80, 13%, p = 0.003), but was not different when compared with age-matched gastric biopsies (44/177, 25%, p = 0.12). CONCLUSIONS: The incidence of H. pylori in patients undergoing RYGBP was higher than that found in all patients undergoing endoscopy and biopsy and than those undergoing fundic biopsies, but not higher when age-matched controls were considered. PMID- 11433902 TI - Laparoscopic vs open Roux-en-Y gastric bypass: a prospective, randomized trial. AB - BACKGROUND: The feasibility of laparoscopic Roux-en-Y gastric bypass (Lap-RYGBP) for morbid obesity is well documented. In a prospective randomized trial, we compared laparoscopic and open surgery. METHODS: 51 patients (48 females, mean (+/- SD) age 36 +/- 9 years and BMI 42 +/- 4 kg/m2) were randomly allocated to either laparoscopy (n = 30) or open surgery (n = 21). All patients were followed for a minimum of 1 year. RESULTS: In the laparoscopy group, 7 patients (23%) were converted to open surgery due to various procedural difficulties. In an analysis, with the converted patients excluded, the morphine doses used postoperatively were significantly (p < 0.005) lower in the laparoscopic group compared to the open group. Likewise, postoperative hospital stay was shorter (4 vs 6 days, p < 0.025). Six patients in the laparoscopy group had to be re-operated due to Roux limb obstruction in the mesocolic tunnel within 5 weeks. The weight loss expressed in decrease in mean BMI units after 1 year was 14 +/- 3 and 13 +/- 3 after laparoscopy and open surgery, respectively (not significant). CONCLUSIONS: Both laparoscopic and open RYGBP are effective and well received surgical procedures in morbid obesity. Reduced postoperative pain, shorter hospital stay and shorter sick-leave are obvious benefits of laparoscopy but conversions and/or reoperations in 1/4 of the patients indicate that Lap-RYGBP at present must be considered an investigational procedure. PMID- 11433903 TI - Does the laparoscopic approach significantly affect cardiac functions in laparoscopic surgery? Pilot study in non-obese and morbidly obese patients. AB - BACKGROUND: Laparoscopy in bariatric surgery represents a modern method generally associated with lower morbidity and mortality, compared with the traditional surgical approach. However, in patients with impaired cardiovascular function, the laparoscopic approach is limited by the potential adverse hemodynamic impact. We assessed the influence of some laparoscopic procedures on selected cardiac functions in significantly obese patients and in subjects with normal body weight, using transesophageal echocardiography (TEE). PATIENTS AND METHODS: Six subjects with normal body weight (mean BMI 25.3 +/- 3.6 kg/m2), and six patients undergoing laparoscopic gastric banding for morbid obesity (mean BMI 45.8 +/- 7.5 kg/m2) were studied. Heart rate (HR), blood pressure (BP), ejection fraction, cardiac output (CO) and transmitral flow were measured. Parameters were recorded at baseline before the operation (BL), after installation of capnoperitoneum (CP), and after positioning the patient for surgery (SP). RESULTS: Compared to BL, CP and SP were characterized by an increase in HR and BP in both groups of patients. As ejection fraction did not change significantly, the HR changes were accompanied by an increase in CO: (BL 5.8 +/- 2.2 l/min, CP 6.5 +/- 2.6 l/min, SP 6.7 +/- 2.7 l/min, p < 0.05 BL vs CP and SP). Transmitral flow parameters did not change significantly. Hemodynamic changes in subgroups with normal body build and in the obese patients were comparable. There was an increase in CO and pressure rate product in obese individuals. CONCLUSIONS: Our results suggest that the hemodynamic response to laparoscopic surgery is characterized by an increase in CO (due to increased HR) and BP. In subjects without a manifest cardiovascular disease, neither systolic nor diastolic performance was significantly affected by the introduction of capnoperitoneum and positioning of the patient for surgery. Similar results were observed in obese and non-obese subjects. Phase II of this on-going study is focusing on impact and safety of laparoscopy in obese patients with known cardiovascular disease. PMID- 11433904 TI - History of Lap-Band: from dream to reality. AB - BACKGROUND: Laparoscopy in severely obese patients is a surgical challenge due to the deep operative field, massive visceral fat, hypertrophic and steatotic liver and inadequate instrumentation. However, performing bariatric surgery by laparoscopy permits a minimally invasive procedure in patients who are usually considered high risk because of their morbid obesity. The challenge was to overcome technical difficulties of laparoscopy in the morbidly obese. METHOD: We needed to develop a new surgical protocol for the gastric approach in severely obese patients. The existing silicone band could not be used for laparoscopy, and a new prototype of the silicone band for laparoscopic use was designed. Because of ethical reasons, we began this work on the animal model. In an animal lab program using pigs, we refined the surgical technique of the laparoscopic approach. A new design of the adjustable silicone band for laparoscopic use was developed. RESULTS: After a 1-year animal lab program and approval by the ethics committee, we performed our first human laparoscopic adjustable gastric banding on September 1st, 1993. CONCLUSION: The development of the Lap-Band from concept to animal lab, ending in clinical application, has been an advance in bariatric surgery. PMID- 11433905 TI - Pregnancies after adjustable gastric banding. AB - BACKGROUND: We evaluated outcome of pregnancies of morbidly obese women who are within the first 2 years after laparoscopic adjustable gastric banding. METHODS: 215 morbidly obese women of reproductive potential (age 18-45 years), who had agreed to remain on reliable contraceptives for 2 years after surgery, were retrospectively analyzed following bariatric surgery. RESULTS: 7 unexpected pregnancies were observed. 5 pregnancies were full-term (3 vaginal and 2 cesarean deliveries). The birth weights ranged from 2110 g to 3860 g. 2 women had first trimester miscarriages. All gastric bands were completely decompressed due to nausea and vomiting, resulting in further weight gain. 2 serious band complications were observed (1 intragastric band migration and 1 balloon defect), which required re-operation. CONCLUSIONS: Pregnancy in morbidly obese women soon after adjustable gastric banding may occur unexpectedly during a period of weight loss. Prophylactic fluid removal from the band eliminates the efficacy of the obesity treatment. Moreover, this cohort shows an increased incidence of spontaneous abortions and band-related complications. PMID- 11433906 TI - Laparoscopic Italian experience with the Lap-Band. AB - BACKGROUND: An increasing number of surgeons with different levels of experience with laparoscopic surgery and open obesity surgery have started to perform laparoscopic implantation of the Lap-Band. METHODS: An electronic patient data sheet was created and was mailed and e-mailed to all surgeons performing laparoscopic adjustable silicone gastric banding (LASGB) in Italy. Patients were recruited since January 1996. Data on 1,265 Lap-Band System operated patients (258 M/1,007 F; mean BMI 44.1, range 27.0-78.1; mean age 38, range 17-74 years) were collected from 23 surgeons performing this operation. RESULTS: Intra operative mortality was absent. Post-operative mortality was 0.55% (7 patients) for causes not specifically related to LASGB implantation. The laparotomic conversion rate was 1.7% (22 patients). LASGB related complications occurred in 143 patients (11.3%). Pouch dilatation was diagnosed in 65 (5.2%), and 28 (2.2%) of these underwent re-operation. Band erosion was observed in 24 patients (1.9%). Port or connecting tube-port complications occurred in 54 patients (4.2%), 12 of whom required revision under general anesthesia. Follow-up was obtained at 6, 12, 18, 24, 36 and 48 months, and mean BMI was respectively 38.4, 35.1, 33.1, 30.2, 32.1 and 31.5. The percentage of patients observed at each follow-up was > 60%. There was no intra-operative mortality and no complication-related mortality, with acceptable weight loss. CONCLUSION: The LASGB operation is safe and effective, and deserves wider use for treatment of morbid obesity. PMID- 11433907 TI - The technical approach in banding to avoid pouch dilatation. AB - BACKGROUND: Adjustable gastric banding results in good weight loss. Nevertheless, some complications may occur, including slipping of the stomach through the band with pouch dilatation. Initially, the Belachew and Cadiere technique was done with the Lap-Band. Afterwards, to minimize proximal gastric pouch dilatation (GPD), we performed the operation using the Swedish route with the same band (Inamed). METHODS: In a retrospective study, 139 consecutive adjustable gastric bands were placed laparoscopically between December 1994 and March 2000. Mean age was 37 years. 10.3% were male. Mean BMI was 39.7. Until April 1999 (Group I, n = 104), the band was introduced according to Belachew's and Cadiere's technique (intragastric balloon calibration technique). Starting May 1999 (Group II, n = 35), the Lap-band was introduced using the Swedish route. This technique consists of localizing the right and left crus posteriorly. A tunnel is created behind the cardia and right above the crus after transsection of the gastrophrenic ligament. The Lap-band is introduced as well as an anterior intragastric calibrating balloon with an air chamber at its distal end, making a pouch 5 to 10 cc. RESULTS: In group I, 15.4% had GPD needing re-hospitalization. Of these, 75% required a re-operation. In group II, no slipping nor pouch dilatation has been reported so far. CONCLUSION: The Swedish route appears to be the key to avoiding GPD. By introducing an intragastric calibrating balloon with a pouch of 5 to 10 cc anteriorly, the band is placed just below the cardia, and no pouch dilatation has been found. The important factor may not be the type of band but rather the technical approach. PMID- 11433908 TI - Laparoscopic gastric banding with Lap-Band for morbid obesity: two-step technique may improve outcome. AB - BACKGROUND: Laparoscopic placement of an adjustable gastric band is an attractive alternative for patients who can benefit from a restrictive bariatric procedure. Creation of the retrogastric tunnel (RGT) may, however, be a considerable challenge early in the surgeon's learning curve. Recent reports described up to 10% band slippage and occasional gastric perforation associated with RGT. The two step (TS) technique involves a crural dissection towards the angle of His through a gastrohepatic ligament approach. It facilitates passage of the band's tubing posteriorly with no wide posterior gastric wall dissection. PATIENTS AND METHODS: Prospective data were registered for the 109 patients (92 females, 17 males) who underwent laparoscopic adjustable gastric banding from December 1998 to May 2000. In 11 patients the standard RGT approach was used, and in 98, the TS technique. The two groups were demographically similar. Mean age was 37 years (18-59); mean preoperative weight was 120 kg (90-165). RESULTS: All procedures were completed laparoscopically. The mean operative time was 59 minutes (31-150) and the mean hospital stay 1.2 days (1-5). Complications in the TS group were gastric wall hematoma in one patient, 3 days of intubation postoperatively in one patient, damage to a band demonstrated in a postoperative contrast study in one patient, and a port-site hernia in one patient. There was no band slippage in the TS group. Among the 11 patients undergoing RGT, there was band slippage in three (27%), immediately postoperatively in one and after 3 and 11 months in the other two. In a mean follow-up of 7 months (1-18), similar weight loss was found in both groups. The mean BMI decreased from 44 kg/m2 (36-61) preoperatively to 40, 38, 36, 34 kg/m2 at 1, 3, 6 and 9 months respectively. 52 patients required band adjustment; of these, 12 required two adjustments. CONCLUSION: Our experience with both the RGT and TS techniques indicates that the latter may offer better results, particularly in the early experience period. It is recommended that in their initial experience with the adjustable band, surgeons should become familiar with this approach. PMID- 11433909 TI - Quality of life after obesity surgery, an evidence-based medicine literature review: how to improve systematic searches for enhanced decision-making and clinical outcomes. AB - BACKGROUND: Research attests to the exponential sweep of clinical obesity in America. Obesity has become one of the most profound public health concerns, closely linked to increased morbidity, mortality, and social, occupational, and psychological discrimination. It is incumbent for specialists to design treatment and outcome analysis based accurately upon systematic research. Regulators, payors, and most notably, patients, need reliable and effective treatment to determine practical standards, manage cost of care, and protocols. METHODS: A comprehensive evidence-based medicine (EBM) review was done of relevant research on quality-of-life (QoL) outcomes after obesity surgery. The Cochrane Collaboration was the EBM searching resource utilized for this project, accessed via the Internet. A systematic EBM search design was implemented. RESULTS: In the search, QoL reported 3,234 hits, "obesity surgery" reported 33 hits, QoL and obesity surgery" reported 6 hits, and QoL and hypertension reported 288 hits. CONCLUSION: We found that a thorough EBM search can be achieved using the Cochrane Collaboration, which provides an efficient, effective and ethical means to enhance evidence-based clinical decision-making in treating severely obese surgery patients. PMID- 11433910 TI - Management of obesity with the new intragastric balloon. AB - BACKGROUND: The introduction of an endoscopically-placed Bariatric Intragastric Balloon (BIB) provided the opportunity to reexamine weight reduction methods and also study potential weight loss without resorting to surgical intervention. METHODS: 10 severely obese patients with mean age 33 years and mean body mass index 39, underwent BIB placement, 7 as a sole weight reduction procedure and 3 requiring weight reduction before repair of large incisional hernias. All patients were followed at 2-week intervals by a nurse practitioner and dietitian for 6 months. RESULTS: Mean weight loss was 18.6 kg (range 6.6-40.0), equivalent to 40% excess weight loss (EWL), range 10-81%. EWL was 54% (29-81%) in those patients who had two balloons placed, who lost an average of 30.3 kg (24.0-40.0 kg). In the patients who had only one balloon placed, mean weight loss was 10.4 kg (8.8-12.5), equal to an EWL of 19% (10-37%). CONCLUSION: These results lead us to consider BIB placement as a successful short-term measure for weight loss or for patients requiring at least weight loss before other surgery. PMID- 11433911 TI - Three years experience with the new intragastric balloon, and a preoperative test for success with restrictive surgery. AB - BACKGROUND: The BioEnterics Intragastric Balloon (BIB) is a smooth, spherical, saline-filled, silicone elastomer with a black radiopaque filling valve, intended to induce weight loss by limiting food consumption. This can be considered a "restrictive" procedure, and by using this balloon, we can assess the patient's eligibility for a restrictive surgical procedure (the BIB-Test). METHODS: From May 1997 to May 2000, 87 BIB were inserted in 77 moderately to severely obese patients (4 BIB in 1 patient, 2 BIB in 7 patients). 64 patients completed the treatment. Out of these, 18 (16 female, 2 male) underwent laparoscopic gastric banding after BIB removal. RESULTS: After the treatment (3-6 months), weight loss results were as follows: WL 14.3 kg, %EWL 23.5 and loss in BMI 5.3. 12 patients after the preliminary BIB, have been followed > or = 6 months after gastric banding, and have significant further weight loss. CONCLUSIONS: BIB appears to have good results. A supervised nutritional and behavioral regimen is mandatory. The balloon may be indicated to: 1) induce weight loss in patients whose obesity is not severe enough to warrant surgery; 2) reduce the surgical risk in those who are massively obese; 3) select patients for gastric restrictive surgery if they lose weight with the balloon. The data showed that patients who had good results with the BIB (positive BIB-Test) are still losing weight after subsequent gastric banding. PMID- 11433912 TI - Canadian occupational therapy research: the little profession that could. PMID- 11433913 TI - Driving and dementia: a review of the literature. AB - In North American society driving is closely linked with independence. Unfortunately, the freedom to operate a motor vehicle may be lost when an individual develops a specific medical diagnosis. The complex issue of dementia and driving safety is frequently encountered by health care professionals. Physicians are required, by law, to report any medical diagnosis such as dementia, that may affect driving safety. Physicians often refer to occupational therapists to assist them in determining if an individual's impairment significantly impacts driving safety. Unfortunately many health care professionals are not using reliable, valid and sensitive tests to determine the point at which an individual with dementia will become an unsafe driver. Through a review of the literature, the authors explore the effects of normal aging and cognitive impairment on driving safety. Specific assessment tools used to assess driving ability are examined and the role of health professionals in driver assessment is discussed. Some suggestions to improve the overall approach to evaluating driving safety are offered in the conclusion. PMID- 11433914 TI - Issues in bidding for contracts for occupational therapy services. AB - There is an increasing number of occupational therapists in Canada who are involved in bidding for contracts to deliver occupational therapy services. Occupational therapists working in an institutional or community-based setting may not have had the responsibility of developing a proposal or a marketing plan for bidding purposes. However, the responsibility of developing a bid to compete for a service delivery contract often rests on occupational therapists who are sole practitioners in a private practice setting. The purpose of this paper is to highlight issues in the literature such as service delivery plans, marketing strategies and costing of services that can assist the occupational therapist in the development of a contractual bid. A specific clinical example, school therapy services, has been used to illustrate how these strategies can be applied to practice. Success in contractual bids appears to be primarily influenced by cost of the service, the expertise of the service provider, ability to provide coordinated care, ease of access for clients, and inclusion of methods to measure client outcome. PMID- 11433915 TI - Postmodernism and its application to the field of occupational therapy. AB - This article presents both the general concept of postmodernism and its reflection in a wide array of fields of interest. In particular, the paper reviews the postmodernist perspective as it appears in healthcare and medicine. This leads to a postmodernist analysis of the profession of occupational therapy, the main conclusion being that occupational therapy combines elements of modernism and postmodernism. This gives occupational therapy clinicians the luxury of enjoying the best of both worlds. PMID- 11433916 TI - [The transformation process among a group of heart patients followed in ergotherapy with the aim of modifying the balance of occupation]. AB - Transformative learning is a process of adaptation that allows for profound personal change in occupational therapy clients. This process is provoked by a triggering factor, such as illness, which results in a critical reflection on values, beliefs, feelings and personal knowledge associated with the object of change (Mezirow, 1991). As part of an objective aimed at modifying the balance of occupation, nine participants were interviewed over a 6 month period to explore their processes of change. Eight concepts related to the objective of change of the balance of occupation emerged from an inductive data analysis process (Glaser and Strauss, 1967). The perception of work was interpreted by the authors as having undergone the most significant sense of transformation amongst the participants. Two other perceptions, 1) the definition of self and 2) the concept of health emerged as new elements essential to the modification of the balance of occupation. The transformation process linked to these eight concepts revealed a specific structure and sequence. The results highlight the primary role of critical reflection amongst occupational therapy clients and underline the importance of deconstructing common values and beliefs which could slow down the process of change. Moreover, the development of new values and beliefs is a vehicle for change in occupational therapy. PMID- 11433917 TI - Intrinsicality: reconsidering spirituality, meaning(s) and mandates. AB - Canadian occupational therapists have placed spirituality as the central core of their theoretical Model, depicting inner and outer selves that contradict simultaneous declarations concerning the integration of mind/body/spirit. Even the word spirituality has discrepant meanings and failure to articulate one chosen meaning leads to ambiguity. This paper argues that occupational therapists must agree upon a single definition of spirituality that is congruent with our professional mandate and philosophical perspective; and that prevention of misunderstandings between and amongst clients and other health care professionals demands recourse to our own terminology. It is proposed that intrinsicality be employed to articulate the personal philosophy of meaning with which we interpret our lives. Influenced by environmental context and in homeostatic relationship with the body and mind, intrinsicality constitutes the essence of the self and informs occupational choices based upon personal values and priorities. Acknowledgement of intrinsicality respects the uniqueness of individuals' meanings. PMID- 11433918 TI - Reflections on reflective practice. AB - In recent years, a wide range of professions have adopted 'reflective practice' as an approach to professional development, and many professions have made it a mandatory dimension of their membership credentialling process. Despite the fact that it has been widely taken up in the professional world, there are many different conceptualizations and ideas about what it is. In this paper, six theoretical underpinnings of reflective practice are considered, and suggestions are made about how we can begin to incorporate reflection into our own practices. PMID- 11433919 TI - [Changes of the incidence and survival in pediatric malignant tumors between 1988 1997, according to the data of the Hungarian Pediatric Cancer Registry]. AB - This paper reports the results of the epidemiological surveillance of childhood malignancies in Hungary from 1988 through 1997, according to the database of the Hungarian Paediatric Cancer Registry. The number of analysed cases was 2146. The crude incidence of all childhood malignancies was 132 per one million person years. The number of new cases diagnosed in Hungary varied between 240 to 280 per year. This number did not change significantly over the observed period in spite of the decreasing number of children in Hungary, therefore, the incidence showed a significant increase of 3.3% per year. The authors also present data about the geographical distribution of childhood cancer in Hungary and survival rates for different tumour types. The 10-year overall survival rate of all malignant diseases diagnosed in Hungary during the analysed ten-year period was 62.6%. PMID- 11433920 TI - [Factors influencing early extubation after open heart surgery]. AB - The authors have performed a retrospective study in order to review the occurrence and the influencing factors of early extubation among their patients. Those patients who had any severe complication in the immediate postoperative period (pericardial tamponade, low cardiac output syndrome, re-operation due to excessive bleeding, postperfusion lung syndrome, pulmonary edema) preventing early extubation, have been excluded from the study. In the remaining 690 patients early extubation within 8 hours and within 4 hours could be carried out in 525 (76.1%) and 164 cases (23.8%) respectively. Late (beyond 12 hours) extubation occurred in 68 cases (9.9%). Anaesthesia was governed by two different methods. Midazolam and alfentanyl (group 1) were used in 137 cases (19.9%) whilst 553 patients (80.1%) received propofol and alfentanyl (group 2). In group 1 and 2 early extubation was possible in 50.4 and 82.5% respectively (p < 0.0001). In further investigations 27 pre- and intraoperative variables of each patient have been studied and analysed. For statistical analysis authors used the SPSS software including T-test, Mann-Whitney-test, chi-square test and multivariate logistical regression analysis. On the basis of multivariate regression analysis factors influencing early extubation were as follows: age (B = 0.0775; p < 0.001), sex (B = 1.2900; p < 0.001), method of anaesthesia (B = 1.9753; p < 0.001), duration of anaesthesia (B = 0.0053; p < 0.001), re-do operation (B = 1.0482; p = 0.0469) and preoperative congestive heart failure (B = 0.9008; p = 0.0125). Pulmonary diseases known from patient history have not had a deep impact on early extubation. On the basis of our study early extubation has not resulted in an increased number of either the postoperative complications or the occurrence of perioperative myocardial infarction. PMID- 11433921 TI - [Clinical characteristics of subcortical arteriosclerotic encephalopathy (Binswanger's disease)]. AB - The subcortical arteriosclerotic encephalopathy (Binswanger's disease) usually starts over 50 years, and it is characterised by slow progression, dementia and stroke-like episodes. The authors analyzed the data of 60 patients, treated in the Neurological Department of Medical University of Pecs between 1994-1997. There was no difference between genders. Among the risk factors only the hypertension and smoking showed correlation with incidence of the disease. The leukoaraiosis found by the CT or MRI was graded according to severity and extension and was correlated with other clinical data (duplex scan, electroencephalographic, and Modified Mini Mental State examination). The morphological damage was related to the age and the decline of cognitive functions, especially with loss of memory, attention and slowing the speech and motor functions regarding to the involvement of the subcortical structures. The role of possible pathogenetic factors and the pathomechanism is discussed. PMID- 11433922 TI - [Prevalence of methylenetetrahydrofolate reductase (MTHFR) gene polymorphism (C677T) in the Hungarian population]. AB - MTHFR encodes a critical enzyme in folate and homocysteine metabolism and the C677T allele of the MTHFR gene has some association with an increased risk for neural-tube defects and for adult cardiovascular diseases. As part of an international collaborative study the prevalence of C677T homozygous genotype was 11.1% while the frequency of C677T heterozygous condition was 45.2% in the Hungarian neonate sample. These findings underscore the clinical importance of the C677T variant in the Hungarian population and urge population-based prevention of conditions related to such gene. PMID- 11433923 TI - [Common bile duct tumor in a young woman with ulcerative colitis]. AB - Bile duct carcinoma is a rare complication of ulcerative colitis. In most of the cases it tends to occur together with primary sclerosing cholangitis predominantly in older males. The authors report a case of a 25 year old woman presenting with jaundice, 6 years after the diagnosis of colitis was made. The cause of the extreme extra- and intrahepatic bile duct dilation was revealed by endoscopic retrograde cholangiopancreatography and percutaneous transhepatic cholangiography showing polypoid tumor in the common bile duct. The histological result taken during the surgical exploration proved the diagnosis of adenocarcinoma. Radical pylorus preserving pancreato-duodenectomy was performed. Subsequently adjuvant chemotherapy was instituted according to the PAV protocol. This rare case proves, that a malignant bile duct tumor may develop in a young patient with ulcerative colitis without the presence of primary sclerosing cholangitis. The authors emphasise the connection between ulcerative colitis and bile duct carcinoma and the importance of the close follow-up of every patient with ulcerative colitis. PMID- 11433924 TI - [Medical History Library--50 years old]. PMID- 11433925 TI - Postoperative beta irradiation for control of pterygium. AB - PURPOSE: The purpose of this retrospective analysis is to assess the effectiveness of postoperative irradiation with a strontium 90 (Sr-90) contact applicator in the prevention of recurrence of resected pterygia. METHODS AND MATERIALS: Thirty five patients were treated with Sr-90 postoperative radiation therapy performed immediately after surgery. The dose of irradiation was 2500 cGy in five fractions using a standard Sr-90 applicator. RESULTS: The local control rate was 94.6% in the first year after surgery. Pterygia eventually recurred in 5.8% of the 52 eyes treated. In our study primarily treated patients remained recurrence free, but recurrences developed in three eyes irradiated after multiple previous excisions. No patient developed conjunctival ulceration, damage to cornea, eye pain, cataract or any other serious complications. CONCLUSIONS: Postoperative irradiation was effective in preventing local recurrence after surgical removal of pterygium. In our experience, acute and late morbidity up to three years is nil. A very satisfactory dose/time/fractionation appears to be of 2500 cGy in 5 fractions starting immediately after surgical excision. PMID- 11433926 TI - Put your (tobacco) money where your mouth is. PMID- 11433927 TI - ECG of the month. The big chill. Hypothermia. PMID- 11433928 TI - Radiology case of the month. Asymptomatic abdominal mass in a child. Wilms' tumor (Nephroblastoma). PMID- 11433929 TI - Louisiana medicine in the mid-1880's. PMID- 11433930 TI - The state of allergic diseases, 2001. PMID- 11433931 TI - Along came an allergen--the science of aerobiology. PMID- 11433932 TI - The inflammatory nature of allergic rhinitis. AB - The importance of the role of mucosal inflammation in the pathophysiology of allergic rhinitis is becoming clearer. Only by addressing the underlying inflammation can adequate control of allergic rhinitis be achieved. Appropriate management of allergic rhinitis is necessary to help control the significant morbidity of this disease, which includes reduced school and work productivity due to absences and cognitive impairment as well as diminished quality of life due to rhinitis symptoms. A review of allergic rhinitis is presented emphasizing the concept of nasal mucosal inflammation in regard to management choices. PMID- 11433933 TI - Allergen immunotherapy. AB - Allergen immunotherapy has been used since the early 1900's for the treatment of allergic syndromes. The efficacy of immunotherapy for the treatment of anaphylaxis caused by the sting of the Hymenoptera class of insects, allergic rhinoconjunctivitis, and the allergic component of bronchial asthma has been clearly demonstrated in numerous well-designed, placebo-controlled trials. Many recent advances in our understanding of the cellular and molecular basis of the allergic response have permitted the development of innovative experimental approaches that very likely will improve the control of the allergic response in the future. PMID- 11433934 TI - Newer therapies for the treatment of asthma. AB - As the pathophysiology of asthma becomes better understood, a specific course of therapy can be directed to the underlying cause of asthma and not just to the signs and symptoms of asthma. This article presents medications available for physicians to use in their armamentarium to prevent the asthmatic patient from experiencing the inconvenience and frustration of repeated asthma attacks. PMID- 11433935 TI - Adverse drug reactions. AB - Adverse drug reactions are a common problem for patients and their physicians. About 6% to 10% of adverse drug reactions are allergic. Obtaining a detailed history is very important. Discontinuation of therapy with a suspect drug is the most helpful diagnostic modality. It is also the treatment of choice. Consultation with an allergist for individualized evaluation and recommendations may be beneficial. PMID- 11433936 TI - [Constructive synergism of regulatory genes expressed in the course of the eye and muscle development and regeneration]. AB - The expression patterns of regulatory genes involved in the formation of the eye in Drosophila and vertebrates during early development were analyzed comparatively. The results demonstrated that, although the compound eyes of invertebrates and the camera eyes of vertebrates markedly differ in their structure and development, they exhibit a striking similarity at the molecular level. This similarity manifests itself in the fact that the homologous regulatory genes ey/Pax, eya/Eya, dac/Dac, and so/Six, which control the early stages of eye development, are expressed in both groups. Not only was synergism shown in the expression of early regulatory genes, but direct interactions of ey/Pax- and so/Six-encoded transcription factors with DNA and protein-protein interactions between nuclear transcription factors encoded by eya/Eya and dac/Dac were also revealed. Transcription factors produced by expressing gene cascades- ey/eya/dac/so in invertebrates and Pax/Eya/Dac/Six in vertebrates--from the transcription complexes that control eye morphogenesis. Paradoxically, the development of muscles in vertebrates proved to involve the expression of genes homologous to the same regulatory genes that control eye morphogenesis in invertebrates and vertebrates. In the developing muscles, regulatory genes also produce transcription factors that form transcription complexes with the mechanism of action based on protein-DNA and protein-protein interactions. The processes of regeneration in the eye and skeletal muscles are controlled by the homologues of the same regulatory genes. Thus, the Pax/Eya/Dac/Six regulatory network is a general system involved in regeneration as well as in development. PMID- 11433937 TI - [Biological model of accelerated senescence. II. Age-related changes in the number of developing male germ cells and Sertoli cells in gonads of senescence accelerated mice]. AB - It was shown that in immature three- to four-week-old mice prone to accelerated senescence (SAMP1 strain), the number of spermatogonia, pachytene spermatocytes, and circular spermatids exceeded that in mice resistant to accelerated senescence (SAMR1 strain) by more than two times. Differences were found in the pattern of age-related changes in the number of meiotic and postmeiotic cells in the sexually mature SAMP1 and SAMR1 mice. In the gonads of SAMP1 and SAMR1 mice, the number of Sertoli cells was unstable. PMID- 11433938 TI - [Chemical basis of nonspecific immunity]. AB - Our knowledge on the nature and quantity of reactive O2 forms generated in phagocytes, particularly, in neutrophil leucocytes, and their role in nonspecific immunity is reviewed. In thermodynamical terms, oxygen is a very reactive molecule and, hence, can react with most chemical elements and many organic molecules. In kinetic terms, O2 is rather inert. Its reactivity can be increased either by reduction or excitation. After accepting four electrons, O2 is finally reduced to H2O. Partial reduction resulting in highly reactive intermediates, namely, superoxide anion (O2.-), hydrogen peroxide (H2O2), and hydroxyl radical (.OH), is possible. Singlet oxygen (1O2) is the product of O2 excitation. Phagocytes acting like agents of nonspecific immunity generate such reactive forms of O2. PMID- 11433939 TI - [Mechanism of cross-linking of fibrinogen and its early structural homolog fragment X]. AB - We studied the mechanism of the cross-linking of fibrinogen as well as its closest structural homolog X fragment under the influence of a fibronectin stabilizing factor (factor XIIIa). The data on elastic and dynamic light scattering indicate the formation of single-stranded polymers without any structural rigidity that acquire a ramified and compact structure upon reaching critical mass. The values of coefficients of translational diffusion, mean-mass molecular weight, averaged scattering factor, and the accumulation of gamma dimers indicate that preincubation of fibrinogen and fragment X solutions significantly accelerates the enzymatic formation of a covalently bound macromolecular protein complex. We propose that enzymatic cross-linking proceeds only with the gradual accumulation of structurally imperfect molecules of fibrinogen and fragment X that are prone to intermolecular D-D end-to-end contacts. PMID- 11433940 TI - [Anticancer properties of flagellate protozoan Trypanosoma cruzi Chagas, 1909]. AB - The anticancer activity of Trypanosoma cruzi has been confirmed by the example of seven strains. Five virulent strains induced the infection, which inhibited sarcoma-180 growth 1.5-22.0 times. The parasites featured tumortropism; i.e., the successfully developed in cancer cells and even preferred them to normal cells. This taxis-based phenomenon was particularly pronounced at cocultivation of the normal and cancer cells. Cultures of the seven (avirulent and virulent) strains can produce an anticancer agent that selectively damages human cancer cells in vitro. The long-term anticancer effect of T. cruzi or preparations from it, as well as possible its cancer preventing effect, has been demonstrated. Three problems are discussed on the basis of the obtained and recently published data: (1) the mechanism of T. cruzi anticancer effect; (2) the nature of the anticancer agent; and (3) the distribution of the considered phenomenon among trypanosomatides. The anticancer activity of T. cruzi may be due to a combination of surface cellular antigens and an inhibiting or lysing factor. PMID- 11433941 TI - [clpP2 gene encoding peptidase in cyanobacteria Synechocystis sp. PCC 6803 controls the sensitivity of cells to photoinhibition]. AB - A homozygous insertion mutant with the inactivated clpP2 gene, which encodes the proteolytic subunit of ATP-dependent peptidase, was obtained in the unicellular cyanobacterium Synechocystis sp. PCC 6803. The mutant cannot grow under photoautotrophic conditions, but cells grown under heterotrophic conditions in a glucose-containing medium have active photosystems I and II (PS I and PS II). The loss of capacity for photoautotrophic growth is determined by a high sensitivity of mutant cells to the inactivating effect of light. Their incubation under light with an intensity above 10 microE m-2 s-1 inhibits cell growth in culture and causes degradation of photosynthetic pigments. It is proposed that the ClpP2 peptidase is involved in the protection of Synechocystis 6803 cells from photoinhibition. PMID- 11433942 TI - [Endogenous abscisic acid level in wheat plants upon inoculation with the powdery mildew causative agent (Erysiphe graminis f.sp. tritici)]. AB - We studied the dynamics of the abscisic acid content in above-ground parts of wheat cultivars with different resistance against powdery mildew. It was assayed by the HPLC method in healthy and diseased (inoculated by the powdery mildew causative agent) plants (from inoculation to the stage of pathogen spore formation). The results are discussed with special reference to the wheat resistance against the powdery mildew. PMID- 11433943 TI - [Molting and seasonal bill-plate shedding in the whiskered auklet (Aethia pygmaea)]. AB - Feather molting and bill-late sheeding were studied because of the unique features of the whiskered auklet biology; i.e., they continue to visit the colony after departure of their young. Like other auklets, the whiskered auklets begin to molt during breeding and do not lose their capacity for flight. The molt pattern of different wing feathers is adaptive and allows new feathers to be protected (when they are soft and could be easily injured) by old or full-grown new feathers during flight or feeding (diving) due to the different timing of the molt of primary feathers and their coverts. The possibility of combining breeding with molt appears to be related to the feeding features of the species. The species that feed on abundant and highly aggregated plankton are able to molt during breeding. The pattern of bill-plate shedding in the whiskered auklet is similar to that in the crested auklet. PMID- 11433944 TI - [Respiration rate and species-specific lifespan in fresh water bivalves of Margaritiferidae and Unionidae families]. AB - We studied changes in the respiration rate in five species of freshwater bivalves as a function of animal age and size. The species-specific lifespan was calculated on the basis of the obtained data: Rubner's constant (Ru) and lifespan at standard temperature 20 degrees C (T20). The longest and shortest lifespan among the studied mollusks was specific for the pearl mussel Margaritifera margaritifera (Ru = 37 kJ and T20 = 36 years) and the freshwater mussel Anodonta anatina (Ru = 8 kJ and T20 = 8 years), respectively. PMID- 11433945 TI - [The opioidergic system in the combined regulation of pain and immunity]. AB - Opioidergic mechanisms are involved in responses to nociceptive and antigenic stimuli at all levels and stages (from peripheral nociceptors to the cerebral cortex and from the precursors of immunocompetent cells to mature effector cells). In most experimental and clinical studies, opioid-mediated analgesia proved to be accompanied by immunosuppression. Opioid receptors of mu, delta, and kappa types are involved in the mechanisms of combined regulation of pain and immunity, with mu and delta receptors suppressing the immune response and kappa receptors enhancing it. By modifying the chemical structure of opioid ligands, it is possible to preserve the analgesic effect and avoid the development of immunosuppression. The opioidergic mechanisms are coupled with nonopioid peptidergic and nonpeptide systems of pain and immunity regulation. PMID- 11433946 TI - [Modeling of values of the net primary production for the zonal vegetation in European Russia]. AB - Several modern approaches to the problem of estimating the net primary production of terrestrial ecosystems are discussed. A method for predicting the dynamics of this parameter as a function of radiation balance and annual evapotranspiration is described. The values of annual carbon by the zonal vegetation of European Russia are calculated in two ways: by methods based on the empirically determined relationships between the annual average values of climatic parameters and net primary production and on the basis of models describing carbon flows between the compartments of ecosystems. The model estimations of net primary production are compared with experimental data. PMID- 11433947 TI - [Classification of the avian summer population of the Western Siberian plain]. AB - The proposed classification system reflects the difference between the three population systems: unbuilt land, built-up land, and aquatic-semiaquatic communities. Two superorder groups--north and median--further divided into types were recognized in each of the systems. Most types are divided into subtypes, classes, and subclasses (and sometimes genera of the population). The estimation of the power and generality of the influence of environmental factors (their variability correlates with heterogeneity of the avian population) has demonstrated that forestation of the territory is most significant in the first half of summer on the western Siberian Plain. The composition of the forest forming species and zoning are less affected. The influence of moisture and hydration is 2-3 times less significant; mesorelief is 4-5 times less significant; and productivity (feeding capacity) and anthropogenic influence are 7-9 times less significant. PMID- 11433948 TI - [Interrelation between the food spectrum of perch fry (Perca fluvatilis L.) and macrozoobenthos structure in experimental mesocosms]. AB - In experimental ecosystems that accommodate perch fry (Perca fluviatilis), significant structural changes within the benthic communities occurred under the effect of fish feeding on oligochaetes and on the larvae of chidonomids and other insects. Perch fry began to consume benthic animals at early developmental stages (C1-C2). The transition to regular feeding under experimental condition occurred at the D2-E stage (days 30-40), when the benthic community reached a developmental peak. The quantitative parameters characterizing the compositions of perch stomach contents and the main groups of zoobenthos closely correlated with each other. The dynamics of quantitative parameters of these animal groups (except for mollusks) proved to be determined both by the specific features of their life cycles and by the degree of consumption. PMID- 11433949 TI - Cholesterol and triglycerides. Numbers to count on. PMID- 11433950 TI - Health tips. Allergic contact dermatitis. PMID- 11433951 TI - New leukemia drug receives FDA approval. PMID- 11433952 TI - Aging of the urinary system. Treatable problems. PMID- 11433953 TI - Dry mouth. Not to be ignored. PMID- 11433954 TI - Wrist fractures. A warning sign. PMID- 11433955 TI - Is breast reduction surgery considered strictly cosmetic? PMID- 11433956 TI - My husband and I just bought cellular phones, but we keep hearing they're dangerous. What are the risks? PMID- 11433957 TI - [Perceptions of quality in health sectors and impact on quality assurance]. AB - According to the ordinances dated April, 24.1996, clinics and hospitals should launch an internal accreditation process. There are many organizations which are about to develop a quality improvement process in order to be prepared to such an evaluation. The success of such approach lies on the involvement of the members of the organization concerned. This survey intends to sort out the actors' perceptions of quality, in order to adapt the process development to the internal cultural context. The data collection was realized through thirty pre-oriented interviews with physicians, nurses, administrative managers and private consultants in charge of implementing the quality process in health structures. The recorded interviews were later on analyzed in order to determine meaningful categories of perceptions of quality. The analysis reveals that quality is perceived in different ways. Three categories of perception can be found: Quality related to "a mean to change the organization" Quality as a "tool to control and limit the expenses" Quality as a way "to improve the relationship with the patients". Administrative managers rather consider the quality process as an expense control tool, whereas physicians and consultants perceive it as a mean to change the organization. Nurses rather adopt the quality process as a way to improve the relationship with the patient. The understanding of how the quality concept is perceived by the different personnel categories helps to initiate appropriately the improvement quality process in services. PMID- 11433958 TI - [Research in general medicine in France: challenges and perspectives]. AB - There is a need to promote research within and on general practice to strengthen the place and prestige of the discipline, in particular within the Universities. This kind of research would contribute to improving the day-to-day work of GP's and consequently, the quality of the services they provide to the patients. The process is under way in all the industrialized countries. However, contradictory forces are crossing the field. The solo practice of most GP's is a barrier to interchange, it slows down the learning process and prevents all kinds of team work, all things necessary to research work. Furthermore, qualitative methods, which are inherent to research in general practice may not be recognized by the scientific community. On the other side, there is a great temptation to reject the conventional standards and proclaim the right to differency, which might be fatal. PMID- 11433959 TI - [The health of developed countries: which is the most cost effective delivery system?]. AB - Several attempts have been undertaken to classify national health care systems according to their merits. The best known is the one published recently by the World Health Organization. The works conducted by the author have produced an other one. Among developed countries, it appears that the model they have adopted -that of Chancelor Bismarck (health services financed by contributions from employers and employees) or that of Lord Beveridge (health services financed by taxes)--does not play a crucial role. Universal access to health services and their provision free of charge do not automatically guarantee equity and efficiency. Equity means that the same treatment is applied to everybody. Efficiency means that health expenditures are kept under control, taking into account the health results obtained. In this respect, the USA are ranked at the lowest level, whatever the classification system used. PMID- 11433960 TI - [General medicine in France and in Germany: current situation and future trends]. AB - The health care system has been built up, both in France and Germany, on the same Bismarckian model: health insurance is financed by contributions from employers and employees, ambulatory care is provided by GPs and specialists in private practice ... etc... Since the 90's, in both countries, there has been a move to strengthen the prestige of GPs and broaden their functions in view of lowering the cost of the health system. However, cost containment has not been the single objective. Several laws have been voted for initiating the gatekeeping function of GPs, controlling the installation of specialists in ambulatory care (private practice), reducing the free choice of care providers by the patients... At the same time, post-graduate training in general practice has been strengthened. Due to historical and cultural reasons, several differences appear in the two countries. For example, confidentiality is strictly observed in Germany and health ID appears only in the French system. We are at the start of a long process. Its success will depend on two conditions: the generalization of the gatekeeping function (which should be entrusted to GPs) and the separation of physicians' income level from the amount of the services they provide to the patients. PMID- 11433961 TI - Chromomeric pattern of maize pachytene chromosomes after trypsin treatment. AB - The bivalent chromomeres of maize were analyzed by means of enzymatic treatment with trypsin. The aim was to verify if the effect of swelling observed in human meiotic chromosomes also occurs in these bivalents, and if this morphological change allows a greater visual resolution of the chromomeric pattern. Bivalents treated with trypsin showed a structure with sequential and linear distribution of small rings. Comparisons between bivalent morphology, treated and non-treated, indicated a collinearity among the sequences of rings and the chromomeres. The results obtained were considered useful to analyze and to make a chromomere map. This treatment can also provide cytological evidence that may help to understand the pairing process. It was concluded that the ring-shape was caused by trypsin induced "disjoining" just in the chromomeric but not in the interchromomeric regions. PMID- 11433962 TI - Chromosome polymorphisms and natural selection in Leptysma argentina (Orthoptera). IV. Survival selection acts on karyotype polymorphisms at the adult stage and before. AB - Deviation from formal null models and the comparisons between age classes are amongst the most direct methods of demonstrating ongoing natural selection. We have sampled 207 males from a population of the grasshopper Leptysma argentina at the onset and the end of the adult life span for this species; generations are synchronical and do not overlap. The karyotypic frequencies for a fusion (F) and a supernumerary segment (s1) were examined. The young male sample was heterogeneous and did not fit to the Hardy-Weinberg proportions, whereas the aged male sample did not deviate from H-W expectations in both analyses. Likewise, the young male sample showed fitness heterosis for the s1 carriers that are fusion homozygotes. We interpreted the significant results of karyotype frequency in both samples as the result of survival selection. Evidently natural selection acts on male adult viability as well as on other fitness components prior to adult survival, thus changing karyotype frequencies between generations. Likewise, we also detected a strong interaction between polymorphisms, whereas heterosis for s1 heterozygotes might be contributing to the maintenance of the s1 polymorphism. The actual target of selection remains unidentified. PMID- 11433963 TI - A new species of Calomys (Rodentia, Sigmodontinae) from Central Brazil identified by its karyotype. AB - Ten species of small rodents of genus Calomys are found in South America. Three of these ten species are known to occur in Brazil: C. tener, C. laucha and C. expulsus (= C. callosus expulsus). Almost all Calomys karyotypes are made up of acrocentric pairs. In this paper we describe a new karyotype with 2n = 46 (FN = 66), including 11 meta/submetacentric and 11 acrocentric autosomal pairs. This is not related to any described Calomys karyotype. The X chromosome is a medium submetacentric and the Y is a small acrocentric. This new karyotype is briefly compared to karyotype of the sympatric species C. tener (2n = 66, FN = 66). The reduced diploid number and small amount of pericentromeric heterochromatin observed in the biarmed chromosomes that contrasts to large blocks seen in acrocentrics seem to indicate that centric fusion and loss of constitutive heterochromatin have led to the new karyotype. Cytogenetic evidence suggests strongly that a new species with 2n = 46 from Central Brazil should be described in the genus Calomys. PMID- 11433964 TI - Cytogenetics of two species of Paratelmatobius (Anura: Leptodactylidae), with phylogenetic comments. AB - In this paper we provide a cytogenetic analysis of Paratelmatobius cardosoi and Paratelmatobius poecilogaster. The karyotypes of both species showed a diploid number of 24 chromosomes and shared some similarity in the morphology of some pairs. On the other hand, pairs 4 and 6 widely differed between these complements. These karyotypes also differed in their NOR number and location. Size heteromorphism was seen in all NOR-bearing chromosomes of the two karyotypes. In addition, both karyotypes showed small centromeric C-bands and a conspicuous heterochromatic band in the short arm of chromosome 1, although with a different size in each species. The P. cardosoi complement also showed other strongly stained non-centromeric C-bands, with no counterparts in the P. cardosoi karyotype. Chromosome staining with fluorochromes revealed heterogeneity in the base composition of two of the non-centromeric C-bands of P. cardosoi. Comparison of the chromosomal morphology of these Paratelmatobius karyotypes with that of P. lutzii showed that the P. poecilogaster karyotype is more similar to that of P. lutzii than P. cardosoi. These cytogenetic results agree with the proposed species arrangements in the P. cardosoi and P. lutzii groups based on morphological and ecological data. PMID- 11433965 TI - A temporal analysis of sexual activity in a natural population of Drosophila willistoni. AB - We have analyzed the mating activity of F1 flies in mating chamber from five samples of wild Drosophila willistoni females. The files were collected from an area (in south Brazil) that has well-defined seasons and considerable variation in temperature and in the abundance of drosophilids. We estimated copulation frequency at first five minutes of observation, copulation frequency at ninety minutes of observation and copulation latency. We did not observe differences in the total copulation frequency throughout seasons, as opposed to copulation frequency in the beginning of the observation period and to copulation latency which varied significantly. The results show that mating activity in a natural population can vary along with the seasons and that this variation can be correlated to some environmental factor. Mating activity tended to be higher during high warm temperature periods, when the flies were abundant. PMID- 11433966 TI - Molecular estimates of primate divergences and new hypotheses for primate dispersal and the origin of modern humans. AB - The concept of recent hominoid divergences has been a mainstay in molecular primatology since the 1970's. However, the ages allocated to the calibration points used to establish these divergence times and the estimates resulting from their application, notably the commonly accepted divergence between Pan (chimpanzees) and Homo 5 million years before present (MYBP), are now palaeontologically refutable. Here we estimate the ages of various primate divergences using three references with a more detailed fossil record than any of the traditional primate calibration points. Our findings suggest that the latter yield datings that are too recent by a factor of about two. For example, our estimates place the divergence between Pan and Homo 10.5-13 MYBP. The revised estimates of primate divergence times suggest a new hypothesis for primate evolution and dispersal: that the divergence between strepsirhines (lorises, lemurs) and anthropoids was contemporary with the break-up of Southern continents about 90 MYBP, with strepsirhines becoming isolated on Madagascar and later dispersing to Africa (and Asia) and anthropoids evolving in South America and subsequently colonizing Africa (and Asia), or possibly North America. In addition we present a new hypothesis, which accommodates the strikingly similar coalescence times for human mitochondrial DNA and the Y-chromosome. This hypothesis posits a common mitochondrial and Y-chromosome bottleneck about 400,000 years ago, associated with the origination of the human 2n = 46 karyotype, obstructing genetic exchange with the 2n = 48 Homo contemporaries. PMID- 11433967 TI - Agrobacterium-mediated transformation of creeping bentgrass using GFP as a reporter gene. AB - Creeping bentgrass (Agrostis palustris Huds.) is a cool season grass widely used on putting greens in golf courses. Transformation of creeping bentgrass has been conducted using microprojectile bombardment and protoplast electroporation. The objective of our study is to develop an alternative and more efficient approach in transforming the grass using Agrobacterium (strain EHA 101). This technique was effective in transforming 40-day old calli derived from mature seeds cultured on MS medium supplemented with 2,4-D, kinetin, and sucrose. Dozens of transgenic plants have been produced from two independent transformed calli. Presence of functional green fluorescence protein (GFP) was detected in leaves, stems, and roots of transgenic seedlings. Four putative transgenic plants and two control plants were randomly chosen and analyzed by Southern blot analysis. Bands corresponding to the GFP gene were clearly shown in transgenic plants. These results indicated that Agrobacterium transformation can successfully be applied to creeping bentgrass. PMID- 11433968 TI - Variability in phenotypic traits in core and peripheral populations of wild barley Hordeum spontaneum Koch. AB - Populations of wild barley, H. spontaneum Koch., were collected in two countries, Israel and Turkmenistan, in environments representing two similar sharp clines of aridity. This allowed us to use the same criteria to define species core and periphery in two regions. Plants from 10 Israeli and 19 Turkmenian populations were grown in a field trial with three water treatments and compared for amount and structure of variation in phenological and morphological traits. Extent of variation was similar in populations at species border (periphery by aridity criterion) or at species border and near it (by habitat), and in populations inhabiting favorable environments away from the border (core). In contrast, two regions (Israel and Turkmenistan) exhibited different amount of variation in phenotypic traits. Israeli populations were more diverse than Turkmenian populations in all except one phenotypic traits and the variation was differently structured among regionally specific core and periphery. While Turkmenian core and periphery defined by either criterion did not differ for all except one trait, corresponding Israeli populations exhibited opposite patterns of variation for trait complexes. Israeli core (Mediterranean) populations were twice as variable than peripheral (desert) populations in parameters of reproductive growth (stem, spike and awn length) and grain filling (spikelet weight) and half as variable in the length of flag and penultimate leaves and onset of reproduction. Possible modes of regionally specific natural selection as a cause of regional/local variation in phenotypic triats are discussed. PMID- 11433969 TI - Chromosome analysis and rDNA FISH in the stag beetle Dorcus parallelipipedus L. (Coleoptera: Scarabaeoidea: Lucanidae). AB - In the present work the chromosome complement (2n = 18; 8AA + XY) of the stag beetle Dorcus parallelipipedus L. (Scarabaeoidea: Lucanidae) is analyzed using conventional Giemsa staining, banding techniques and ribosomal fluorescent in situ hybridization (rDNA FISH). rDNA FISH remains the unique tool for providing a clear-cut identification of Nucleolar Organizer Regions (NORs) when conventional banding methods such as silver- and CMA3-staining proved to be inadequate. The dull, homogeneous CMA3 fluorescence of all chromosomes indicates the absence of markedly GC rich compartmentalized regions in D. parallelipipedus genome. Silver impregnation inadequacy in detecting NOR regions is to be sought in the unusual extensive silver stainability of heterochromatic material which, on the contrary of what stated for vertebrates, seems to be a common feature in Scarabaeoidea species. PMID- 11433970 TI - Chromosomal rearrangement in autotetraploid plants of Arabidopsis thaliana. AB - Recent development of cytogenetic techniques has facilitated significant progress in Arabidopsis thaliana karyotype studies. Double-target FISH with rRNA genes provides makers that allow individual chromosome in the genome to be distinguished. Those studies have revealed that the number and position of rDNA loci is ecotype-specific. Arabidopsis is believed to be a true diploid (x = 5) with numerous ecotypes (accessions) and only a very few natural polyploid populations reported. Few studies were undertaken to induce polyploidy in Arabidopsis, however none of those gave the cytogenetic characteristics of polyploid plants. Our analysis of chromosome pairing of colchicine-induced autotetraploid Arabidopsis (Wilna ecotype) revealed preferential bivalent pairing in PMCs (pollen mother cells). In order to attempt to explain this phenomenon, first of all more detailed cytogenetic studies of autopolyploid plants have been undertaken. The localization of 45S and 5S rDNA loci in the diploid and autotetraploid plants revealed that Wilna ecotypes belongs to the group of Arabidopsis accessions with only two 5S rDNA loci present in a genome. Furthermore, the rearrangement of 45S rDNA locus in autopolyploid, when compared to the diploid plants of the same ecotype, was revealed. These results are interesting also in the context of the recently emphasised role of polyploidy in plant evolution and speciation. Arabidopsis, despite having small chromosomes, is a good system to study chromosome behaviour in relation to diploidization of autopolyploids and to evaluate the degree of chromosomal rearrangements during this process. PMID- 11433971 TI - Spontaneous sister chromatid exchanges in chicken embryos. PMID- 11433972 TI - [No need to question the placebo effect!]. PMID- 11433973 TI - [Warning against a too one-sided evidence-based medicine! Good medicine implies also knowledge of human nature and intuition]. PMID- 11433974 TI - [Both major breakthroughs and stagnation in urology. Industry, together with clinicians, are responsible for most innovations]. AB - Urology is a specialty with many branches, including urological oncology with 25% of all cancers. Development in certain areas been very rapid, for instance with the introduction of minimally and non-invasive methods such as ESWL and phosphodiesterase inhibitors, innovations which have brought obvious improvements and have been promptly adopted in clinical practice. In other areas such as chronic abacterial prostatitis and renal cancer, progress has been very limited. Still other areas have seen useful but less spectacular improvements for which it has taken time, clinical experience and a multitude of clinical studies before they have been embraced in daily clinical practice. Examples of these more gradual developments are hyperthermia for the treatment of benign prostatic hyperplasia and transrectal ultrasound in prostate cancer. PMID- 11433975 TI - [CSF-analyses in clinical diagnosis of Creutzfeldt-Jakob disease. A literature review and three cases from routine clinical practice]. AB - The clinical diagnosis of Creutzfeldt-Jakob disease (CJD) is often difficult, since the clinical presentation varies between patients and is often uncharacteristic. Therefore, CJD is often not suspected until late in the disease process, due to the rapid progress of the disease. Suspected CJD is, however, frequent in routine clinical practice, in the investigation of patients with rapidly progressing dementia and/or uncharacteristic neurological symptoms. We present results from cerebrospinal fluid (CSF) analyses in CJD, focusing on the two neuron-specific proteins 14-3-3 and tau. Analysis of 14-3-3 protein is performed by Western blotting, in which 14-3-3 is either detectable in CSF or not, while CSF-tau is analyzed using quantitative ELISA methodology, in which a markedly increased CSF level of tau (< 1500 pg/mL) is indicative of CJD. Tau and 14-3-3 findings show comparable sensitivity and specificity, higher than the presence of spike and wave complexes on an EEG. The increase in CSF-tau in CJD is substantially higher than in Alzheimer's disease, with low overlap between the disorders. Instead, false positive results are found in disorders with massive acute neuronal damage (encephalitis, stroke and CNS tumors). In cases in which these disorders cannot be differentiated from CJD on clinical grounds, they can often be identified by MRT or by the finding of blood-brain barrier damage or signs of inflammation upon CSF analysis. CSF-tau is increasingly used in the routine diagnosis of Alzheimer's disease, and the analysis also seems to be of use in the clinical diagnosis of CJD. We also present three cases from routine clinical practice, all with a marked increase in CSF-tau, which was the only positive objective sign in two of the cases. PMID- 11433976 TI - [Unbroken continuity of patient care in shoulder pain. Same initial care in primary health services as in hospitals]. AB - Thirty percent of all patients that consult their primary care physicians for pain in their musculoskeletal systems have pain in the neck/shoulder region, while 23% have lower back pain. Of major concern in Sweden is a patient's lengthy waiting period before seeing a specialist, with a resultant delay in treatment. A multidisciplinary group of physicians and physical therapists has therefore been working on and refining a system of tools to provide an "unbroken chain of care" for the diagnosis and treatment of patients with shoulder pathology. An unbroken chain of care enables the clinician regardless of specialty to provide the highest level of care to all patients with the same initial diagnostic and treatment parameters. Axelina is the tool that provides all the standardized treatment algorithms, rehabilitation plans and detailed home exercises. Axelina also contains a two-day instructional package with case studies presented on video. PMID- 11433977 TI - [Psychological aspects of aging--a review]. AB - A number of models of aging have identified predictors of longevity, well-being and life quality among the elderly. The predictive patterns are multifactorial, and in all models psychological aspects are important. Most of all it is a matter of personality, intellectual functioning, adaptive capacity and coping, as well as making use of one's life-spain experiences. Personality, by consensus deemed to be essentially stable all through life, exerts a constant influence on the individual for better or for worse, steering the individual toward excessive wear and tear or constructive development and maturity. Some aspects of intellectual functioning are more important than others in this context, e.g. crystallized and fluid intelligence and episodic short term memory. Crystallized intelligence usually remains unchanged throughout life and even improves with age, while fluid intelligence and episodic memory decline. In very old age as for instance in centenarians, fluid intelligence is found to be a very strong predictor of future survival. Among the elderly in general, terminal decline is evident shortly before death. Coping strategies and capacity combined with type of incidents, change or threat are found to be strong predictors of both life satisfaction and survival after 80 years of age. Identifying the importance of coping in old age has brought about a change in perspective in caring for the elderly in which the psychological aspects become more important as a complement to the medical model. Increasingly, autobiographical reflections or life-story telling, also when used in group sessions, is shown to be one way of increasing well-being and life satisfaction among the elderly. PMID- 11433978 TI - [Female genital mutilation--a complex phenomenon]. AB - As of today, more than 130 million girls and women worldwide are genitally mutilated. Female genital mutilation (FGM) exists primarily on the African continent (along and north of the equator), but the practice is also carried out elsewhere. Clitoridectomy was performed in medicine both in the United States and in Europe as late as in the 1950s. From a socioeconomic perspective FGM is understood as a practice that forms an important part of girls' socialization. FGM is practiced in societies where women's social acceptance and survival is secured through marriage and childbearing. Without FGM, the woman cannot get married, with the consequence that she has no status or access to resources. Her body practically becomes her only form of capital. From a psychosexual perspective FGM is interpreted as a practice that has been made possible in patriarchal societies where the sexuality of women has to be controlled and where unequal gender relationships are preserved. FGM is practiced by people from various religious denominations, among them Copts, Animists, Catholics, Protestants and Muslims, something that goes against the relatively common belief that FGM is practiced only among Muslims. The resistance to FGM found in countries where the practice is rife clearly demonstrates that patriarchal structures and what gets defined as "culture" are indeed possible to change and renegotiate. PMID- 11433979 TI - [Female genital mutilation meets Swedish health care. Female genital mutilation is one of many forms of discrimination of women in the world]. AB - About 27,000 women from countries in which female genital mutilation (FGM) is a common practice are presently living in Sweden. This means that FGM is a phenomenon that directly affects the Swedish health care system. Knowledge and understanding of the background, meaning and consequences of FGM are a prerequisite for effective prevention, proper clinical handling and supportive reception of the women. To avoid a stigmatizing reception it is also important to understand the situation of genitally mutilated women, and to become aware of the identity crisis many of them experience when they come to Sweden and lose their identity as "normal" women. It is essential to remember that female genital mutilation is one of many forms of discrimination affecting girls and women around the world. This discrimination knows no national or cultural borders and varies in expression and extent. In order to offer optimal care and reception of women who have been socialized into a gender role that is often seen as completely different from the gender role that Swedish society is said to embrace, it is of the utmost importance to first take a critical look beneath the veil of alleged gender equality of Swedish women. PMID- 11433980 TI - [Measuring errors in neonatal randomized controlled trials. Deficient quality concerning the informed consent]. PMID- 11433981 TI - [The significance of control for juvenile delinquency]. PMID- 11433982 TI - [The world's mental health day, April 7, 2001. All countries must invest in public mental health]. PMID- 11433983 TI - [How do we help couples with hereditary diseases who want to start a family?]. PMID- 11433984 TI - [Clinical social medicine--yes, please!]. PMID- 11433985 TI - [Final reply: the Gillberg group should be more self critical]. PMID- 11433986 TI - [Taking photos of the accident? We mostly agree]. PMID- 11433987 TI - [Evidence-based diabetology and patients' self care--patient education is more than just blood glucose self-monitoring]. PMID- 11433988 TI - [Basic training in ultrasonographic diagnostics--already a reality in Linkoping]. PMID- 11433989 TI - [Marat might have suffered of chronic myeloproliferative disease]. PMID- 11433990 TI - [TNF-inhibitors slow down joint destruction in rheumatoid arthritis. No cure, but dramatic relief of symptoms]. PMID- 11433991 TI - [Not proved that depleted uranium has negative effects on health]. PMID- 11433992 TI - [Rapid recovery after colonic resection. Multimodal rehabilitation by means of Kehlet's method practiced in Vasteras]. AB - By using a multimodal rehabilitation program professor Kehlet has shown accelerated recovery after colonic surgery with hospital stay of only two days, irrespective of open or laparoscopic technique. These results have not been confirmed in other studies. The aim of this study was to replace our traditional approach with Kehlet's multimodal regimen and try to reproduce his reported data. METHODS: 22 patients (median age 67 years) underwent right- or leftsided colectomies, 15 open (7 with midline incisions) and 7 laparoscopically. Continuous thoracic epidural, immediate mobilization and oral nutrition were used. Discharge was planned three days after surgery. On the first postoperative day all had oral intake and on the third day patients were mobilized for a median of 9.7 hours and all had resumed defecation. Pain and fatigue scores (VAS) were low. The median post-operative hospital stay was 3.5 (range 3-8) days. Two patients returned with complications. No cardiopulmonary or infectious complications were seen. The multimodal rehabilitation programme resulted in a quick recovery and a hospital stay of three days in most patients after colonic surgery. PMID- 11433993 TI - [Small changes in health differences in the Nordic countries during the 1980s and 1990s]. AB - The study examined changes over time in health inequality in Denmark, Finland, Norway and Sweden. Data derive from comparable interview surveys carried out in 1986/87 and 1994/95. Limiting long-standing illness and perceived ill health were analysed regarding age, gender; educational attainment, and employment status. Age adjusted prevalence rates were calculated. Changes in differences in health were found in education and employment status groups. There was little or no change in the prevalence of ill health during the time period studied. Despite social and economic changes differences in health remained broadly stable in the examined countries. PMID- 11433994 TI - [Beer, wine, spirits and mortality]. AB - A population based cohort study investigates the association between alcohol intake and mortality from all causes, coronary heart disease and cancer. The design is prospective with baseline assessment of intake of beer, wine and spirits, smoking habits, educational level, physical activity, and body mass index and a total of 257,859 person-years follow-up on mortality. A total of 4,833 participants died, of these 1,075 from coronary heart disease and 1,552 of cancer. Compared with non-drinkers, light drinkers who avoided wine, had a relative risk of death from all causes of 0.90 (0.82-0.99) and those who drank wine had a relative risk of 0.66 (0.55-0.77). Heavy drinkers who avoided wine were at higher risk of death from all causes than were heavy drinkers who included wine in their alcohol intake. Wine drinkers had significantly lower mortality from both coronary heart disease and cancer than did non-wine drinkers (p = 0.007 and p = 0.004, respectively). In conclusion, wine intake may have a beneficial effect on all cause mortality that is additive to that of alcohol. This effect may be attributable to a reduction in death from both coronary heart disease and cancer. PMID- 11433995 TI - [Holistic perspective is needed in care of multiimpaired elderly. A proposal for structural change of the organization of care in the county of Stockholm]. AB - Studies in the County of Stockholm reveal a malfunctioning care system for elderly persons with one or more diseases in conjunction with immobility and reduced strength. These multi-impaired elderly consume a substantial proportion of care and rehabilitation services. They would need a health co-ordinator to monitor the entire care process, dealing with all aspects of care, and whose sphere of responsibility is not unduly restricted. The communication of information from inpatient care to the primary care setting is highly problematic. Therefore it is proposed that the transfer of multi-impaired elderly individuals from inpatient care to outpatient primary care be a carefully monitored process. In addition, outside of the hospital an "elderly team" that would be composed of doctors, district nurses, rehabilitation personnel and care assistants is proposed. This team should have a geographically well-defined area of responsibility, in which they would provide care in the homes of these multi impaired elderly. PMID- 11433996 TI - [Health praising coated in olive oil]. PMID- 11433997 TI - [Discussion of the revision of the Helsinki Declaration is needed]. PMID- 11433998 TI - [Modernism characterized by vision phenomenon. "Blind spots" in Kandinsky's art express changed consciousness]. PMID- 11433999 TI - [Forensic psychiatry a resource in crime prevention]. PMID- 11434000 TI - [Reformed forensic psychiatry in patients' and users' perspective]. PMID- 11434001 TI - [Yielding claim adjustment important factor behind the increase of whiplash injuries]. PMID- 11434002 TI - [Lack of respect for pediatric neuropsychiatry as a field of knowledge]. PMID- 11434003 TI - [Epidemiology and mercury]. PMID- 11434004 TI - [Lack of organ donors--reverse the problem!]. PMID- 11434005 TI - [Ernst Schweninger. Physician with a "doubtful reputation", who tamed Bismarck, was distrusted by his colleagues but loved by his patients]. PMID- 11434006 TI - [Mannheim Conference of the 24th Congress of the European Association of Pediatric Cardiologists, Strasbourg, June 2000. Evolution and complexity of the relationship between pediatric cardiologists and life]. PMID- 11434007 TI - [Magnetic resonance imaging of dilatation of the ascending aorta after repair of coarctation of the aorta]. AB - The authors studied the risk factors for dilatation of the ascending aorta in patients operated for coarctation of the aorta. A prospective study of the diameters of the ascending aorta by magnetic resonance imaging was undertaken in 46 patients with an average age of 30 months (range 6 days to 11 years) at surgery, and 10 years of age (6 months to 31 years) at the time of the investigation. The diameters were measured at the level of the sinus of Valsalva, at the sino-tubular junction, and compared with reference tables with respect to body surface area. Twenty six per cent of patients had dilatation of the ascending aorta. The predisposing factors were investigated. Age, type of surgery, postoperative hypertension. Doppler gradient in the isthmic region, anatomical appearances of the repair observed by MRI were not predictive of this complication. On the other hand, age of patients at MRI and bicuspid aortic valves (present in 66% of cases) (p < 0.05) were significant risk factors. These results indicate that regular follow-up by echocardiography or MRI of the diameter of the ascending aorta is necessary in patients operated for coarctation of the aorta and with bicuspid aortic valves. PMID- 11434008 TI - [Baloon angioplasty with stent implantation in recoarctation of the aorta: an attractive alternative]. AB - The results of transcatheter balloon angioplasty in teenagers and adults with aortic recoarctation are uncertain. Therefore, there is a current trend to prefer a more complex procedure including the implantation of a stent. This study deals with 8 patients aged 7 to 25.3 years (median: 15 years), weighing 20 to 68 kg. (median: 57) and having undergone resection of an aortic coarctation during infancy (24 days to 4 years). All had their lower limb pulses diminished or abolished, elevated blood pressure at rest (and at exercise in the 5 tested patients), and left ventricular hypertrophy. MRI documented the lesion and helped to select seven patients whose stenosis was short and remote enough from the origin of the main aortic collateral. In one case, the decision to stent was taken as an emergent measure to treat an aortic dissection which appeared shortly after balloon dilatation. The effectiveness of the procedure was immediate in all patients with a 50% increase in diameter of the dilated area, total relief of the gradient, drop to normal values of the blood pressure. These good results persisted at follow-up (3-24 months) in 6 patients, with moderate hypertensive rebounds in the last 2. There were 2 technical problems (premature burst of the balloon, asymmetrical inflation of the stent like an "Eiffel Tower") that could finally be overcome and should no longer occur with the new specially designed so called "BIB" balloons. Would long term follow-up confirm these early results, one should conclude that this method offers an attractive, safe and effective option to surgery for adolescents and adults with late recoarctation of the aorta. PMID- 11434009 TI - [Enlarging angioplasty of the aortic arch with an pulmonary artery autograft patch. Interest in a single-stage repair of the syndrome of coarctation and interruption of the aortic arch]. AB - Single stage repair of syndromes of coarctation and interruption of the aortic arch is a routine procedure in many surgical centres with good immediate results. The classical technique of aortic repair is based on the principles of Crafoord's extended resection anastomosis. Recoarctation is not an unusual long-term complication. A technique of enlarging angioplasty of the aorta using a patch of pulmonary artery has been developed and used in 22 neonates with obstruction of the aortic arch associated with ventricular septal defect with an average age and body weight of 15 days and 2.9 Kg respectively. The ventricular septal defect was closed surgically during the same procedure. Total circulatory arrest was not used in these children and all had aortic repairs with selective cerebral perfusion with moderate hypothermia (28-30 degrees C). This technique was used without any procedure-related early morbidity. No early or late deaths were observed in this series. Two patients were reoperated during the first year after the initial procedure: one for residual ventricular septal defect and the other for supraventricular pulmonary stenosis. Two patients, one of whom was reoperated, developed supraventricular pulmonary stenosis with a gradient of over 60 mmHg. These stenoses were observed in the first cases operated and were essentially due to the technique of pulmonary artery reconstruction. Over a median follow-up period of 10 months, no recoarctations were observed: the Doppler ultrasound study showed an isolated mean systolic gradient of 6 +/- 12 mmHg. The authors conclude that angioplasty of the aortic arch with an enlarging patch of pulmonary artery autograft during single stage surgery of syndromes of coarctation and interruption of the aortic arch provides a harmonious and durable repair of the aortic arch. PMID- 11434010 TI - [Percutaneous closure of patent ductus arteriosus with the Amplatzer duct occluder. Results of 29 patients]. AB - The authors report their experience of percutaneous closure of large patent ductus arteriosus with the Amplatzer duct occluder. The age of the patients ranged from 3 months to 72 years (average 24 months) with body weights of 3.9 to 74 Kg (average 16 Kg). The smallest angiographic diameter of the ductus was 4.0 +/- 1.3 mm at angiography (range 1.8 to 6 mm). The occluder was inserted under local anaesthetic with control aortography in all but one case who required general anaesthesia. In two small babies, the implantation was performed after venous puncture alone with transthoracic echocardiographic monitoring. The implantation was successful in all but one case in which there was a distensible duct. Early angiographic control showed suppression of the shunt in 61% of patients. Secondary haemolysis was observed in one case and was corrected by a second catheterisation 4 days later, with occlusion of the duct with a balloon catheter. The patients were followed up clinically, radiologically and echocardiographically. Doppler echocardiography showed absence of a residual shunt in 24 patients (86%) one month after the procedure, in 25 patients (89%) 3 months after the procedure and in 26 patients (93%) one year after the procedure. Two patients were left with mild or moderate shunts 12 and 16 months after the attempted occlusion. No cases of aortic or pulmonary obstruction were observed. The authors conclude that the Amplatzer duct occluder is a reliable device for closing large patent ductus arteriosus. It may be proposed in young symptomatic children over 4 Kg in body weight. A high rate of occlusion is obtained with a minimal risk of complications. PMID- 11434011 TI - [Rehabilitation of hypoplastic pulmonary arteries and anatomic correction of pulmonary atresia with interventricular communication]. AB - Conventional treatment of pulmonary atresia with ventricular septal defect (VSD), hypoplastic pulmonary arteries (PA) and major aorto-pulmonary collaterals (MAPCAs) is controversial: from symptomatic and palliative treatment for some authors to surgery with unifocalisation of collaterals for others. These treatments never use native pulmonary arteries as only source of pulmonary flow, but create "neo-pulmonary arteries". Nine cases of pulmonary atresia with VSD, hypoplastic PA and MAPCAs were treated by rehabilitation of native PA through a staged approach: 1) surgical neonatal connection between right ventricule (RV) and hypoplastic PA, 2) evaluation and interventionnal catheterism with angioplasty of PA stenosis and closure of collaterals, 3) complete surgical correction with reconstruction of right outflow track and PA and closure of VSD. After first surgical stage of RV-PA connection at the mean age of 4.8 months (+/- 5.6 months), 8 patients were alive and underwent 22 cardiac catheterisms (mean of 2.7 per patient), with angioplasty of PA, and occlusion of MAPCAs in 6 and 2 patients respectively. Seven patients underwent complete anatomical correction at the mean age of 28.8 months (+/- 17.7 months) with one late death. The 6 remaining patients had encouraging hemodynamic status (RV pressure/LV pressure ratio at 0.6 +/- 0.26; mean left and right distal pulmonary pressure at 15.2 mmHg (+/- 9.1 mmHg)), and good functionnal status (3 in NYHA functionnal class 1, and 3 in class 2), for a mean follow-up of 79.5 months (+/- 41.4 months). One patient had reoperation on right outflow track stenosis, 6 years after correction. This small series enhances the feasibility of a staged approach with rehabilitation of small PA, allowing complete surgical correction with the native PA with good hemodynamic and functional results in pulmonary atresia, with VSD, hypoplastic PA and MAPCAs. PMID- 11434012 TI - [Interest in myocardial scintigraphy following the arterial switch procedure for transposition of the great vessels]. AB - Coronary artery obstruction is the main late complication of the so-called arterial switch operation designed to repair transpositions of the great arteries in newborn infants by switching the great vessels and transferring the coronary ostia onto the posterior vessel. Our aim was to study the links between myocardial perfusion and coronary artery anatomy after the arterial switch operation. Forty-five patients (5.863 years) underwent a 201Tl myocardial SPECT and a selective coronary artery angiography. The latter was normal in 20 children: 13 had also a normal myocardial scan but 7 had myocardial perfusion defects including 2 with angina who had a very low coronary reserve at positron emission tomography. Twenty-five patients had severe coronary artery lesions: 5 with a normal myocardial scan and 20 with perfusion defects. Twelve out of these 20 underwent surgical revascularization and the SPECT images went back to normal in all within 6 months after surgery. Specificity and sensitivity of myocardial SPECT in detecting coronary artery lesions were 78% and 69% whereas positive and negative predictive values were 74 and 73%. We conclude that myocardial SPECT imaging is not the right way to detect late post arterial switch coronary artery lesions. It is helpful in decision making as to submit these children to surgical revascularization and in assessing its postoperative effectiveness. PMID- 11434013 TI - [Short and medium-term results of the Norwood intervention (first stage)]. AB - We report the short and mid-term results of the Norwood procedure (Stage one) in 20 patients with hypoplastic left heart syndrome or univentricular heart with aortic obstruction. Seven patients were prenatally diagnosed. Preoperatively there was obstruction to pulmonary venous return in 6 cases, a mild to moderate tricuspid regurgitation in 6 cases, and 11 patients were supported by mechanical ventilation with multiorgan failure in 5 cases. The surgery was performed under cardiopulmonary bypass at a mean age and weight of 12.9 days and 3 kg, respectively. Nine patients (45%) died within 30 days postoperatively, whereas 5 had delayed sternal closure. The mean duration of mechanical ventilation and ICU stay were 5.7 and 11 days, respectively. Two patients were reoperated for bronchial compression and tracheotomy. Systemic venous thrombosis occurred in 5 patients. In multivariate analysis, an older age at surgery was correlated with postoperative hospital death (p = 0.03). Among the 11 patients discharged home after Stage one procedure, 5 patients underwent balloon dilation for recoarctation and one patient died at home. A bidirectional cavopulmonary anastomosis was performed in 8 patients at a mean age of 0.76 year, with one postoperative death. After a mean follow-up of 1 year (+/- 1.97 years), the 9 remaining patients are all in NYHA class I, at a mean age of 2.2 years. Their mean transcutaneous saturation is 81%. The Norwood procedure (Stage one) is associated with high hospital mortality. However, the functional status of the survivors is correct, like in patients with other type of univentricular hearts. Moreover, although the causes of death in our patients are often not clarified, other studies show that the leading causes of deaths in our patients are often not clarified, other studies show that the leading causes of death in those patients are correctable. In conclusion, the option of a Norwood procedure (Stage one) should be proposed in patients with hypoplastic left heart syndrome (or variant). PMID- 11434014 TI - [Variation in cholesterol, lipoproteins and triglycerides after heart transplantation in children]. AB - Total cholesterol, HDL and LDL-cholesterol and triglyceride levels may contribute to the development or progression of coronary artery disease of the transplanted heart. The aim of this retrospective study was to determine the short and long term lipid profiles of transplanted children and to identify factors influencing these dyslipidemias. Twenty-three patients aged 9.5 +/- 5.9 years at cardiac transplantation were followed up for 5.8 +/- 3.1 years. All were on triple therapy with normal diets. The total cholesterol increased by 17% during the first year (4.47 +/- 1.01 mMol/l to 5.25 +/- 1.22 mMol/l at 1 year: p < 0.05) with a peak at 3 months of 5.31 +/- 1.28 mMol/l correlating with the dosage of prescribed corticosteroids. LDL-cholesterol levels increased by 20% during the first year (2.26 +/- 0.67 mMol/l to 3.29 +/- 0.99 mMol/l at 1 year: p = 0.018). HDL-cholesterol levels increased from 1.02 +/- 0.27 mMol/l to a maximum of 1.55 +/- 0.4 mMol/l at 1 year, p < 0.05. Lipoprotein A1, a protecting sub-fraction of HDL, did not change significantly. Changes in triglyceride levels were not significant despite a tendency to hypertriglyceridaemia in the early phases. After one year, serum cholesterol and lipoprotein levels remained higher than the initial values. These results show that cardiac transplant children are exposed to the risk of atherogenic hyperlipidaemia and require systematic lipid profile monitoring, dietary advice and lipid lowering drugs. PMID- 11434015 TI - [T wave abnormalities on Holter monitoring of congenital long QT syndrome: phenotypic marker of a mutation of LQT2 (HERG)]. AB - The two genes which code for the potassium channels, KCNQ1 and HERG, are responsible for the most common forms of the long QT syndrome, LQT1 and LQT2. Abnormalities of duration and morphology of the ventricular repolarisation are amongst the diagnostic criteria of this syndrome. The morphology of the T waves was studied by 24 hour Holter monitoring in 190 subjects with a long QT syndrome due to KCNQ1 (LQT1) [N = 133] or HERG (N = 57) and in 100 controls, and it was compared with the ECG T wave. The T wave was characterised according to 3 morphological features: grade 0 (G0) = normal, grade 1 (G&) = slight ST depression and grade 2 (G2) = presence of ST elevation of the descending phase of the T wave. The T wave morphology on Holter ECG was normal for most LQT1 and control subjects compared with LQT2 (92%, 96% and 19% respectively, p < 0.01). Grade 1 appearances were observed more often in LQT2 (18 vs 8% for LQT1 and 4% for controls, p < 0.01). Grade 2 appearances were only observed in the cases of LQT2 (63%). The predictive factors of G2 were young age and an anti-sense mutation of the transmembrane domaines of HERG. The authors conclude that Holter monitoring improves detection of T wave changes compared with the ECG. Grade 2 changes seem to be a phenotype marker for a HERG mutation, especially those situated in the transmembrane domaines. PMID- 11434016 TI - [Socio-professional acclimation of children operated on for cardiac disease]. AB - In order to assess the socio-professional insertion of children operated for cardiac disease, 177 patients over 20 years of age completed a questionnaire about their scholastic achievements, training and socio-professional categories. This formation was compared with that of the French population based on the statistics of the "Institut national de la Statistique et des Etudes Economiques" (INSEE). The cardiac patients were more likely to have technical occupations than the general population and their level of scholastic achievement reflected this tendency. They attained better professional situations than commonly thought. The analysis of the conditions of surgical treatment indicated that a single, curative and early operation is the best guarantee for future achievements. However, the foundation of a family is much more difficult for operated cardiac patients than for the general French population. Failure of socio-professional insertion is far from being a fatality and psychological problems play an important role, implicating the practitioner at three levels: patient, family and society. In order to be totally convincing, the study of the future of operated patients requires analysis of many different factors. PMID- 11434017 TI - [Quality of life of adults with univentricular hearts]. AB - This study sought to evaluate the quality of life of surviving adults with univentricular heart and to determine the impact of sociodemographic and clinical characteristics on quality of life. The health records of 89 survivors with univentricular heart living in France at the time of the study were reviewed. They were 17 to 49 years old (median: 21). Sixty-seven patients answered the Duke questionnaire. Sociodemographic and clinical variables were similar in the responders and non-responders. The scores of patients who answered the questionnaire were compared with national norms. The impact of sociodemographic and clinical variables on individual Duke's measures was assessed. The Duke scores of adults with univentricular heart were similar to those of the normal population. Cyanosis predicted a worse score for physical (p = .05) and perceived health measures (p = .02). The higher educational level predicted a better score for physical (p .004), mental (p = .01), and general health measures (p = .02). Orthopaedic problems worsened social score (p = .05). Psychosocial problems worsened pain score (p = .04). Mitral atresia, in comparison with the other anatomical types, worsened perceived health score (p = .02). Finally, patients younger than 23 years scored better for almost all of health and dysfunction measures. In conclusion, despite repeated interventions and other disease-related everyday stresses, a selected group of adults with univentricular heart had a satisfying quality of life. These patients probably developed copying mechanisms, easing the psychological stress, and oriented themselves towards a different set of values in everyday life so that to accept disabilities and to recalibrate personal expectations. PMID- 11434018 TI - [Cardiac consequences of adolescent anorexia nervosa]. AB - Cardiac complications are common in adolescent anorexia nervosa and are the cause of a third of deaths. Some workers have reported prolongation of the QT interval and cases of sudden death in these patients. The aim of this study was two-fold: to assess the cardiac complications of anorexic adolescents and to determine the outcome after renutrition in the hospital setting. This was a prospective study of 48 consecutive cases (45 girls) with an average age of 14 +/- 2 years, admitted to the paedopsychiatric unit and fulfilling the DSM-IV criteria of anorexia nervosa. The digitised ECG, Holter ECG and echocardiography were recorded before and after renutrition. Anorexia nervosa was severe with a body mass index < 14 in 2/3 of cases. Over 2/3 of patients had bradycardia with a heart rate < 50/min in half the cases but normal chronotropic function on Holter monitoring. Prolongation of the QTc interval was demonstrated (QTc > 440 ms in 11/44 cases). Echocardiographic abnormalities, in particular left ventricular dysfunction (24/46) and pericardial effusion (12/46) were reversible after renutrition. There were no clinical or biological predictive factors for the occurrence of cardiac complications on admission. The authors confirm that cardiac complications of anorexia nervosa are common, usually benign and always reversible after renutrition in hospital. Therefore, most electrical abnormalities normalise with the heart rate and echocardiographic abnormalities with improvement of conditions of load. PMID- 11434019 TI - [Myocardial infarction in a child with an anomalous left coronary artery arising from the right coronary sinus. Value of echocardiography]. AB - An anomalous left coronary artery arising from the right sinus of Valsalva with a trajectory between the aorta and the main pulmonary artery is a rare isolated congenital anomaly carrying a high risk of sudden death and of acute myocardial ischaemia, especially in children and young adults in a context of exercise. It is usually a post-mortem finding. The authors report the rare case of a 12 year old child who suffered acute myocardial infarction on exercise due to this condition. The diagnosis was made in the acute phase by echocardiography. Surgical correction was performed as this is the only means of prevention of sudden death of an ischaemic recurrence. Effort syndromes in children or young adults should lead to a request for echocardiographic examination to exclude the diagnosis in a non-invasive manner as well as those of hypertrophic cardiomyopathy or congenital aortic stenosis. PMID- 11434020 TI - [Left monoplegia revealing pulmonary arteriovenous fistual in an adolescent: occclusion with an Amplatzer occluder]. AB - A 16 year old adolescent was admitted to hospital for investigation of a transient left monoplegia. The clinical, electric and radiological examinations were all normal. Echocardiography showed no abnormality of cardiac structures or function. Contrast echocardiography was performed and showed a right-to-left shunt suggesting a pulmonary arteriovenous fistula which was confirmed at angiography. The fistula was closed at catheterisation by a detachable balloon: in a second procedure, the balloon was deflated and contrast studies showed a recurrence of the right-to-left shunt. An Amplatzer 8/6 duct occluder was implanted with excellent results and no recurrence at 3 months. PMID- 11434021 TI - [Ross mitral procedure for massive congenital mitral insufficiency]. AB - The authors report the case of a 4 month old North African baby with massive congenital mitral insufficiency associated with a coarctation of the aorta. The surgical treatment consisted in correction of the coarctation followed, several days later, by a Ross mitral procedure after failure of mitral reconstructive surgery. The postoperative course required circulatory assistance for 4 days and the child returned to his family two months after surgery. Control echocardiography before hospital discharge showed good function of the pulmonary autograft in the mitral position with no regurgitation but slight stenosis with a mean transvalvular pressure gradient of 4 mmHg. This procedure may be used in certain cases as an alternative to mitral valve replacement. PMID- 11434022 TI - [Medullary extradural hematoma revealing a coarctation of the aorta]. AB - The authors report a spontaneous, unusual complication of coarctation of the aorta. An 11 year old child was admitted for investigation of chest pain. Cardiovascular examination revealed typical clinical signs of coarctation of the aorta. Neurological examination found neck stiffness without headache or deficit. The presumptive diagnosis of dissection of the aorta was infirmed by echocardiography and MRI. The latter investigation, with views of the spinal cord, revealed a compressive medullary extradural haematoma. Antihypertensive therapy and corticosteroids with strict bed rest resulted in complete regression of the haematoma and the coarctation was operated 6 months later. Medullary complications of coarctation of the aorta are usually postoperative. Spontaneous complications are exceedingly rare but very serious: medullary compression by the dilated anterior spinal artery or rupture of an aneurysmal collateral vessel. In this case, magnetic resonance imaging led to diagnosis and effective early treatment of this complication before the patient developed a neurological deficit and the coarctation was treated surgically thereafter. PMID- 11434023 TI - [Mitral valve prolapse and type II mucopolysaccharidosis. Report of two familial cases]. AB - Type II mucopolysaccharidosis (Hunter's disease) is a hereditary condition due to a deficit of a lysosome specific hydrolase (iduronate sulfatase) inducing an accumulation of dermatane-sulphate and heparane-sulphate in certain organs. Cardiac involvement is constant in this disease and manifests itself essentially by aortic valve stenosis and insufficiency and/or mitral insufficiency which is progressive, irreversible and life-threatening. Two brothers of Portuguese nationality suffering from a so-called slight form of this disease had classical mucopolysaccharide infiltration of their aortic valves. The elder brother, aged 11, had severe aortic insufficiency associated with mild stenosis requiring treatment with vasodilator drugs. The younger, aged 8, had asymptomatic mild aortic regurgitation. Curiously, mitral valve prolapse with regurgitation was present in both children. The association of mitral valve prolapse and type II mucopolysaccharidosis, without other typical cardiac involvement, has only been reported once in the literature. PMID- 11434024 TI - [Antenatal atrial tachycardia. Two case reports]. AB - The authors report two cases of foetal supraventricular tachycardia in healthy hearts with 1/1 atrioventricular conduction which turned out to be atrial tachycardias in the postnatal period. The first foetus had permanent tachycardia at 190/minute at 34 weeks' amenorrhea with left ventricular dysfunction at 36 weeks. In the postnatal period, treatment with digoxine and amiodarone restored sinus rhythm and normal left ventricular function. Permanent foetal tachycardia, even at a rate of less than 200 beats/minute, should suggest an arrhythmia and may lead to left ventricular dysfunction in utero. The other foetus had an aneurysm of the foramen ovale with paroxysmal tachycardias at 220/minute without cardiac dysfunction. A Holter at 1 month showed paroxysmalatrial tachycardia. Postnatal rhythm monitoring is necessary in paroxysmal foetus tachycardia, especially with prenatal aneurysm of the foramen ovale. PMID- 11434025 TI - [Value of an implantable EKG monitor for the diagnosis of arrhythmic syncope in children]. AB - The value of an implantable ECG monitor (Reveal TM Plus) is reported in a 12 year old child with unexplained syncopal episodes despite extensive investigations. Twenty-seven days after the implantation, ventricular tachycardia at 450/min was recorded at the first recurrence. This case shows that this type of monitoring can be particularly useful when an arrhythmia is thought to be the cause of syncope in a child and the initial investigation is negative. PMID- 11434026 TI - [A rare cause of cardiomyopathy in childhood: propionic acidosis. Three case reports]. AB - Dilated cardiomyopathy is a rare and unrecognised complication of propionic acidosis. It usually progresses to chronic cardiac failure with a poor prognosis. The authors report three cases of cardiomyopathy due to this condition. One of the children underwent liver transplantation which cured the cardiac complication. PMID- 11434027 TI - Analytical perspectives of cooperative coastal management. AB - We accept the thesis that coastal management, as any other form of environmental management, can be effective only with the cooperation of a multitude of stakeholders with conflicting interests. At present, cooperation is forced upon stakeholders by a paternalistic (top-down) coastal management that is outcome oriented and coercive in nature. Forced cooperation is difficult to maintain, however. The alternative is to seek voluntary cooperation with a process oriented, cooperative (bottom-up) coastal management approach. After a brief review of these arguments, we address the major analytical challenge of cooperative coastal management, which is to search for solutions that can be negotiated and implemented with maximum voluntary cooperation. The main property of these solutions, which are also referred to as core solutions, is that they are preferable to individual stakeholders or coalitions of stakeholders over acting-alone alternatives. Our analysis is applicable to any other form of environmental management. PMID- 11434028 TI - Contaminated sites: the Canadian situation in an international context. AB - Over the last 2 decades, policy-makers have been giving increasing attention to the remediation and redevelopment of contaminated sites, especially those located in urban areas commonly referred to as brownfields. Traditionally, private developers have tended to ignore these sites on account of a series of obstacles of a structural-political nature, including variability in regulatory processes, lack of information on soil quality, impractical clean up standards, fear of liability, and limited funding resources for clean ups. This paper examines the types of policy-making measures that are currently being taken in Canada to overcome these obstacles, comparing them to those being taken in the US and Europe. It is argued that the contaminated site-related policies and programs employed to overcome each obstacle, both within Canada and internationally, are converging in style and content as governments are becoming more aware of the types of costs and risks they must share in order to solve the problem effectively. It is also argued that this trend is unfolding in a relatively predictable way, and that policy-making in Canada is evolving more slowly than it is in the other jurisdictions examined. PMID- 11434029 TI - Degradation of endosulfan in a clay soil from cotton farms of western Queensland. AB - The persistence and degradation of endosulfan isomers and their primary degradation product, endosulfan-sulfate, were studied in a clay soil from cotton farms of western Queensland. Endosulfan degradation in relation to soil moisture, temperature, day and night temperature fluctuation, waterlogging and re application were studied. The results show that the degradation rates of both endosulfan isomers were greatly affected by changes in soil water content and temperature. Under a high water content-high temperature regime the concentration of alpha-endosulfan in the soil fell rapidly during the first 4 weeks of application, followed by a prolonged period of slower rate of degradation. Alpha endosulfan showed a bi-exponential form of degradation for all water content temperature experiments except for extremes in both these two factors. In the submerged soils (and also in low-water content, low temperature, non-submerged experiments) no such rapid initial degradation of alpha-endosulfan was observed, and a single first-order rate equation describes the data. Degradation of beta endosulfan was significantly slower than for the alpha-isomer under all conditions studied. A half-life of more than a year was recorded for the beta isomer when both water content and temperature were low. The degradation of beta endosulfan showed no sign of the bi-exponential function observed for alpha isomer, and a single first order rate equation described the data obtained for each factor studied. Endosulfan-sulfate was the major degradation product in all non-submerged experiments. Its build up in the soil very closely followed the disappearance of alpha-endosulfan. Its highest build-up was in the high water content-low temperature experiments, but its persistence was primarily influenced by soil temperature. Both alpha and beta-isomers, and endosulfan sulfate, persisted longer in the submerged soil. Re-application of endosulfan, and day and night fluctuation of temperature had contrasting effects on the degradation of the two isomers. Both factors slowed down the degradation of alpha-endosulfan and enhanced that of beta-endosulfan, but their net effect was to prolong the overall persistence of this chemical in the soil. Submerged conditions reduced the net formation of endosulfan-sulfate and enhanced its degradation rate. PMID- 11434030 TI - An evaluation of the predictive performance of distributional models for flora and fauna in north-east New South Wales. AB - To use models of species distributions effectively in conservation planning, it is important to determine the predictive accuracy of such models. Extensive modelling of the distribution of vascular plant and vertebrate fauna species within north-east New South Wales has been undertaken by linking field survey data to environmental and geographical predictors using logistic regression. These models have been used in the development of a comprehensive and adequate reserve system within the region. We evaluate the predictive accuracy of models for 153 small reptile, arboreal marsupial, diurnal bird and vascular plant species for which independent evaluation data were available. The predictive performance of each model was evaluated using the relative operating characteristic curve to measure discrimination capacity. Good discrimination ability implies that a model's predictions provide an acceptable index of species occurrence. The discrimination capacity of 89% of the models was significantly better than random, with 70% of the models providing high levels of discrimination. Predictions generated by this type of modelling therefore provide a reasonably sound basis for regional conservation planning. The discrimination ability of models was highest for the less mobile biological groups, particularly the vascular plants and small reptiles. In the case of diurnal birds, poor performing models tended to be for species which occur mainly within specific habitats not well sampled by either the model development or evaluation data, highly mobile species, species that are locally nomadic or those that display very broad habitat requirements. Particular care needs to be exercised when employing models for these types of species in conservation planning. PMID- 11434031 TI - Fishing and farming at Lake Chad: institutions for access to natural resources. AB - Lake Chad is a vitally important wetland in the semi-arid Sahel corridor. It provides the basis of many thousands of livelihoods which depend on its seasonal fluctuations to renew fish stocks, farmland and rangeland. This paper describes how access to farmland and fishing rights has evolved on the Nigerian shore of the lake. The paper aims to assess the applicability of different institutional approaches to natural resource management on the lake shore. Although many recent approaches to natural resource management have reflected a 'Tragedy of the Commons' approach, a growing literature both in support and critical of Hardin's (1968) thesis has followed. Four distinct approaches are considered: (1) institutional intervention to prevent 'tragedy'; (2) institution erosion brought about by such interventions; (3) models of institutional adaptation to resource scarcity; and (4) approaches which perceive institutions as a crucial determinant of social and economic development. In examining which institutional approaches may be relevant at Lake Chad, it is anticipated that this paper will provide insights which will be relevant to policy-makers, resource managers and students working in environments where resource fluctuation inhibits stable property rights and state resource management has proved neither feasible nor successful. The western shore of Lake Chad has been under the jurisdiction of Borno State in its various guises since the end of the fourteenth century and is currently one of 36 states in the Federal Republic of Nigeria. Although the administrative status of Borno itself has varied, it has been dominated by a Kanuri aristocracy for most of its existence. The Kanuri administration has continued to operate in a remarkably similar way over the past 150 years. Traditionally, it has played a crucial role in allocating access to farm land. In recent years, the 'Kanuri administration' has not only maintained its pre-colonial authority over farming on the lake shore, but has expanded it to cover new areas of the lake floor and the increasingly lucrative fishing opportunities which federal government has been unable to regulate. This success suggests that collaboration with the organisations which operate such institutions could be beneficial, if not essential, to the success of natural resource management. PMID- 11434032 TI - A profile of recyclers making special trips to recycle. AB - The aims of the research were to: (1) establish a profile of 'special recyclers', i.e. those who make special trips by car to recycle their household waste; and (2) establish whether the environmental burden resulting from their activities presents a significant environmental problem than needs pro-active management. The research was undertaken through surveys of recyclers at supermarket recycling centres and at civic amenity sites in Glasgow and across Ayrshire in south-west Scotland. Twenty-two percent of civic amenity recyclers made special trips to recycle, whilst at supermarkets less than 10% made special journeys. Those making special journeys tended to make shorter journeys than those combining recycling with another activity. Special recyclers also tended, on average, to recycle more items and bring greater weights of recyclable material per trip. No demographic difference was found between the special recyclers and the 'non-specials'. The environmental impact of consumer journeys to recycle was estimated from the statistic: 'The total distance travelled by car by special recyclers/the total weight deposited by all recyclers'. For the sites of this survey, this statistic ranged in value from 9 km tonne-1 to 106 km tonne-1. The lowest impacts of 9 km tonne-1 were at a supermarket site in a small town with a predominantly local catchment. The highest impacts were at a civic amenity site, in a major town, also used by residents from satellite communities. Over 50% of special recyclers cited nearness as their prime reason for choosing a particular site, compared with 32% of non-specials. Although special recyclers in Ayrshire did appear to act reasonably responsibly (environmentally), their activities produced additional environmental burden. Simulation studies demonstrate that up to 34% reduction in this burden may be possible through encouraging longer intervals between recycling trips. Three quarters of the environmental burden was attributable to less than 5% of the overall recycling population. PMID- 11434033 TI - Changing environmental strategies over time: an empirical study of the steel industry in the United States. AB - This study investigates how environmental strategies change over time. We submit evidence from the US steel industry that firms have modified their strategies over time. We offer that US industry passed through three stages--cost minimization, cost-effective compliance, and beneficial environmental controls. We compare typologies of environmental strategies and choose that of C. Oliver as the most appropriate. We investigate how environmental strategies in the steel industry changed over time a 4-year period. We offer that a further understanding of Oliver's strategies may increase understanding of the relationship between business and government on environmental issues. One over-arching problem in our field is the need to adequately operationalize how firms change strategies and pass through different stages. We hope that our study will help future researchers and practitioners better articulate the concepts of environmental strategies over time. Our study focused on the steel industry in the United States. We chose the US steel industry as one of the major environmental actors in the United States. The United States Environmental Protection Agency ranks the iron and steel industry as the largest industrial source of toxic environmental contamination. We encourage researchers to evaluate and test our methodology and findings in other contexts--both in other nations and different industries. PMID- 11434034 TI - Reply to Huber. Estimating Markov transitions. PMID- 11434035 TI - Partnership for front-line success: a call for a national action agenda on workforce development. AB - Despite more than a decade of dialogue on the critical needs and challenges in public health workforce development, progress remains slow in implementing recommended actions. A life-long learning system for public health remains elusive. The Centers for Disease Control and Prevention and the Agency for Toxic Substances and Disease Registry in collaboration with other partners in federal, state, local agencies, associations and academia is preparing a national action agenda to address front-line preparedness. Four areas of convergence have emerged regarding: (1) the use of basic and crosscutting public health competencies to develop practice-focused curricula; (2) a framework for certification and credentialing; (3) the need to establish a strong science base for workforce issues; and (4) the acceleration of the use of technology-supported learning in public health. PMID- 11434036 TI - A systems approach to public health workforce development. AB - During the 1990s, several distinct but interrelated efforts to strengthen the public health infrastructure were launched. Defining public health work in terms of core functions and essential services, these efforts focused on the competence of the workforce and the performance of public health agencies. The systems approach offered here highlights the relationships and interdependencies among these three components of public health practice: (1) the work, (2) the worker, and (3) the work setting. The model suggests that advances in public health workforce development may require major public health organizational development efforts. PMID- 11434037 TI - Assessing the professional development needs of public health professionals. AB - This article describes the major activities associated with designing and implementing a comprehensive, professional development needs assessment of public health professionals in four states of the South Central region of the United States. The instrumentation, research design, and summary results of the needs assessment described in this article may facilitate similar efforts by interested researchers and program developers to assess the public health professional workforce training needs. Results of needs assessments can be useful in designing and evaluating professional development curricula and activities to strengthen public health services in the United States. PMID- 11434038 TI - The credentialing of a population-based health profession: lessons learned from health education certification. AB - This article summarizes research and issues related to health education certification. Health education's experience with certification provides valuable insight for other public health professions because of the credential's population-based emphasis, its recent development, and its broadly defined cross cutting skill set. Although data are still emerging on the outcomes of health education certification, most studies indicate that it has had a positive impact across multiple dimensions. Critical to this success has been providing profession-wide opportunities for input, communicating with key stakeholders, establishing an independent credentialing agency, and basing the credential on scientifically and legally defensible competencies that are linked to job responsibilities. PMID- 11434039 TI - Professionalizing the public health workforce: the case for certification. PMID- 11434040 TI - Chasing the technology rabbit. PMID- 11434041 TI - Joint Council of State and Local Health Officials: workforce development- principles for action. PMID- 11434042 TI - A conceptual model for leadership development. AB - Collaboration among schools of public health and national, state, and local health agencies has resulted in creation of comprehensive public health workforce education and training initiatives that offer integrated, sequential, and accessible professional development programs, including a nation-wide network of public health leadership institutes. A conceptual model for leadership development is presented. It contains seven elements considered critical for design of leadership programs in public health: capacity/competence needs; program target; area served; program content; training level; learning approach; and implementation methods. This model can be used to design leadership as well as public health workforce education and training programs. PMID- 11434043 TI - National Public Health Performance Standards: workforce development and agency effectiveness in Florida. AB - The Florida Department of Health (FDOH) was the first state to pilot test both the Centers for Disease Control and Prevention (CDC) state agency and local Public Health System Performance Standards. The standards were found to be complementary and supportive of the FDOH quality performance improvement system, which had been in place for a decade, and the new Quality Management initiative. The pilot test found uneven performance across the state's county health departments and identified several areas, especially workforce development, that required additional efforts. The FDOH, in collaboration with the Center for Leadership in Public Health Practice at the College of Public Health in the University of South Florida, have collaborated and will continue to collaborate to design and deliver training in critical workforce development areas. PMID- 11434044 TI - Competency-based credentialing of public health administrators in Illinois. AB - This article describes an initiative to develop and implement a competency-based credentialing program for public health managers and administrators that is linked with practice performance standards for local public health systems. The Illinois Public Health Administration Certification Board represents an innovative model for credentialing public health workers, placing equal value on competencies secured through education and training and those demonstrated in practice. Competency-based credentialing of public health administrators may have applicability for other segments of the public health workforce. PMID- 11434045 TI - Enumeration of the public health workforce: developing a system. AB - Although it has been of interest to public health leaders, advocates, and policy planners for many years, decades have passed since the last organized count of public health workers. This article reports on methods used by the Columbia University School of Nursing, Center for Health Policy, to enumerate the public health workforce in 57 states and territories based on existing reports, summaries, and surveys. The complexity of public health workforce data is described and the scheme utilized to characterize the workforce using public agency categories is illustrated. The resulting "best current estimate" provokes many questions regarding future policy about a public health workforce database. PMID- 11434046 TI - Building the analytic capacity of the State Maternal and Child Health workforce- a history of the HRSA/MCHB Academic Partnership. AB - The Maternal and Child Health Bureau of the Health Resources and Services Administration (HRSA) in conjunction with Schools of Public Health has sponsored a variety of graduate education and continuing education initiatives during the last 15 years aimed at enhancing the analytic capacity of the maternal and child health (MCH) workforce. These initiatives are described, with lessons learned as well as recommendations for future efforts provided. PMID- 11434047 TI - Training readiness of public health agencies: a framework for assessment. AB - The Healthy People 2010 Objectives and the Taskforce on Public Health Workforce Development both have recognized the important roles of state and local agencies in the education and training of their employees. This article describes an initial step in the development of a tool to assess agencies' training readiness using five factors derived from learning organization theory. These factors (resources, policies, learning culture, programs, and leadership) offer a useful framework for further development of a tool to assess training program readiness. PMID- 11434048 TI - Improving performance at the local level: implementing a public health learning workforce intervention. AB - In an effort to continually improve performance of the essential public health services with community partners, the diverse public health workforce in a major metropolitan area engaged in an organizational learning process. Core public health organizational competencies, identified in a multi-year collaborative applied research initiative, provided the curricula content for the public health learning experience. All members (about 600) of the Columbus and Franklin County (Ohio) Health Departments participated in four one-half day small group, highly interactive modules conducted during a 2-year period. The purpose of this article is to describe the design and implementation of this workforce intervention, the lessons learned, and implications for developing organizational capacity and improved performance. PMID- 11434050 TI - Prehospital diagnostics used to reduce time to treatment in patients with acute coronary syndromes. PMID- 11434051 TI - Milestones in the development of the first chest pain center and development of the new Society of Chest Pain Centers and Providers. PMID- 11434052 TI - Acute coronary syndromes: introducing new therapies into established guidelines. AB - Quantitative (or semiquantitative) risk stratification is becoming a more accurate and feasible way to define both overall risk and likelihood of benefit from more potent and invasive therapies. LMWH appears to be at least as good, and likely modestly better than, unfractionated heparin, and when used for patients undergoing revascularization, it should be continued until revascularization. GP IIb/IIIa inhibitors are highly effective for coronary intervention, as well as for "up-front" use in high-risk ACS patients in an environment in which intervention is commonly used. In spite of the new guidelines, a clear delineation to the practitioner of which treatments are proven effective, in which patients, and in what combinations, remains challenging. Best estimates are that only 50% to 75% of ideal patients are currently receiving proven therapies. Better methods are needed to fully integrate proven effective treatments into practice. PMID- 11434053 TI - Acute coronary syndrome care: emergency department strategies. PMID- 11434054 TI - Attack programs in chest pain centers. PMID- 11434055 TI - Chest pain centers in Brazil. PMID- 11434056 TI - The killer chest pain. PMID- 11434057 TI - Technology and risk stratification: tools for decision making. PMID- 11434058 TI - Electron beam computed tomography in chest pain centers. PMID- 11434059 TI - Observation units for elimination of missed myocardial infarction errors. PMID- 11434060 TI - Designing safe systems of care and their application in chest pain centers. PMID- 11434061 TI - Prodromal symptoms of acute myocardial infarction: overview of evidence. PMID- 11434062 TI - Cyclic coronary flow: defining preinfarction angina at the crossroads of unstable angina and myocardial infarction. PMID- 11434063 TI - How to set up a chest pain center--the ABCs. PMID- 11434064 TI - The changing reimbursement pathways in emergency department chest pain centers for patients with acute coronary syndrome. PMID- 11434065 TI - The Early Heart Attack Care (EHAC) program. A tool for saving lives. PMID- 11434066 TI - Cardiopulmonary resuscitation: strengthening the links in the chain of survival. PMID- 11434067 TI - Good, better, best "infarctus interruptus" view from a public health cardiologist's perspective. PMID- 11434068 TI - Hastening the coming of the science of peace. PMID- 11434069 TI - The effects of combined thrombolytic and platelet inhibitor therapy on myocardial and microvascular perfusion. PMID- 11434070 TI - Emergency department perfusion imaging for suspected coronary artery disease: the ERASE Chest Pain Trial. PMID- 11434071 TI - Cardiac time-insensitive predictive instruments (TIPIs): real-time emergency clinical decision aids and tools for the retrospective assessment of care. PMID- 11434072 TI - Biochemical markers and the era of troponin. PMID- 11434073 TI - Impact of reducing antibiotic prescribing for acute bronchitis on patient satisfaction. AB - CONTEXT: Using a patient and clinician educational intervention, we successfully reduced antibiotic use for uncomplicated acute bronchitis. The impact of this intervention on patient satisfaction is not known. OBJECTIVE: To evaluate whether a strategy for reducing antibiotic use in acute bronchitis affects satisfaction among adult patients. DESIGN: Telephone survey administered 1 to 4 weeks after an office visit for acute bronchitis. SETTING: Two outpatient clinics belonging to a group-model HMO in the Denver, Colorado, metropolitan area. The intervention clinic had received a patient and office-based educational intervention that successfully reduced antibiotic prescribing for acute bronchitis during the previous winter. The control clinic received only the office-based materials, an intervention that did not reduce antibiotic prescribing. OUTCOME: Overall satisfaction with the episode of care. RESULTS: Antibiotics were prescribed to 64% and 85% of survey respondents at the intervention (n = 102) and control clinics (n = 164), respectively (P < 0.001). Patient satisfaction with the visit did not differ between intervention and control clinics (69% of intervention and 63% of control clinic patients reported very good or excellent satisfaction, P > 0.2). After adjustment for patient age, sex, duration of illness before the visit, reason for visit, and clinician specialty, there was no difference between intervention and control clinics in the proportion of patients reporting very good or excellent satisfaction (adjusted relative risk for high satisfaction at the intervention clinic, 1.1 [95% CI, 0.81 to 1.3]). CONCLUSION: A patient- and clinician-oriented educational intervention that reduces antibiotic treatment of adults with uncomplicated acute bronchitis does not appear to reduce satisfaction with care. PMID- 11434074 TI - Educating men about prostate cancer screening. A randomized trial of a mailed pamphlet. AB - CONTEXT: Although evidence-based guidelines recommend that physicians inform men about prostate cancer screening, the most efficient way to do this is not known. OBJECTIVE: To evaluate whether a mailed educational pamphlet affected men's knowledge about early detection of prostate cancer. DESIGN: Randomized, controlled trial. SETTING: Primary care clinic of the Minneapolis VA Medical Center. PATIENTS: 342 men at least 50 years of age who responded to a mailed survey (overall response rate, 68%) and did not report a history of prostate cancer. INTERVENTION: "Early Prostate Cancer" pamphlet mailed to patients in the intervention group 1 week before their scheduled clinic appointments. OUTCOME MEASURES: Patients' responses to a survey mailed 1 week after their clinic appointments; prostate-specific antigen (PSA) testing determined from electronic medical records. RESULTS: Respondents were predominantly elderly white men (mean age, 71 years; 90% white) with chronic illnesses (48% described their health as "fair" or "poor"). Men who received the educational pamphlet were better informed than men in the usual care group, as measured by correct responses to the following three questions about prostate cancer screening: the natural history of prostate cancer (32% vs. 24%; P = 0.10), whether treatment lengthens lives of men with early prostate cancer (56% vs. 44%; P = 0.04), and accuracy of PSA testing (46% vs. 27%; P < 0.008). The overall proportion of correctly answered questions was greater in the intervention group (45% vs. 32%; P < 0.001). Testing for PSA in the year after the index clinic appointments did not differ significantly between the intervention group and the usual care group (31% vs. 37%; P > 0.2). CONCLUSIONS: Male veterans are poorly informed about the potential benefits and risks of prostate cancer screening. Although our mailed educational pamphlet enhanced knowledge only modestly, it was an inexpensive and easily implemented intervention. PMID- 11434075 TI - Variation in clinical preventive services. AB - CONTEXT: Preventive services are not delivered at optimal rates in primary care settings, and the literature suggests that a systems approach is key to improvement. Studying variation among clinics could help us to understand the extent of system use in practice. PRACTICE PATTERN EXAMINED: The proportion of patients who are up-to-date for preventive services in 44 primary care practices in the Midwest. PREVENTIVE SERVICES EXAMINED: Papanicolaou (Pap) smear, cholesterol testing, mammography, clinical breast examination, blood pressure measurement, influenza and pneumococcal vaccinations, and advice on tobacco use. DATA SOURCE: 6830 patients surveyed after their clinic visit (response rate, 85%). RESULTS: The proportion of patients up-to-date for preventive services varied widely among clinics. For example, up-to-date rates for Pap smear testing ranged from 70% to 93% and 45% to 88% for cholesterol screening. There was little correlation between a clinic's performance on one preventive service (relative to the other 43 clinics) and its performance on others. When correlations between pairs of up-to-date rates within clinics were examined, only 4 of 28 service pairs were positive and statistically significant and only 1 had a correlation coefficient that exceeded 0.5 (for mammography and clinical breast examination). CONCLUSION: There is wide variation in the rates at which various preventive services are performed, both between and within clinics. This variation, which is probably due to a lack of organized prevention systems that cover multiple services, provides a clear target for improvement efforts. PMID- 11434076 TI - Web-based assistance for physicians caring for children with ADHD. AB - CONTEXT: The clinical management of children with attention deficit/hyperactivity disorder (ADHD) requires measurement of behavioral outcomes at home and at school. This information is difficult to obtain. GENERAL QUESTION: Could a Web site that facilitates exchange of information between parents, teachers, and health care professionals improve behavioral outcomes for children with ADHD? SPECIFIC RESEARCH CHALLENGE: Designing a pragmatic intervention and evaluation strategy. PROPOSED APPROACH: Establish a secure Web site to share observations on a child with ADHD. Access to a child's electronic record will be limited to team members defined by the parents. The record will contain documentation of consent for team members to communicate both diagnostic and treatment information. The record will feature two methods of communication: 1) the ability to input and graphically view structured behavioral questionnaire data that measure levels of attention and hyperactivity/impulsivity, and 2) an electronic bulletin board that allows users to post and respond to text messages that specifically describe behavior. The Web site would be tested in a randomized trial. UNANSWERED QUESTIONS: Given its complexity, which part of the intervention should be isolated for study? What is the appropriate unit of analysis (individual, practice, or school)? Can an unbiased assessment of behavior be obtained when parents and teachers are not blinded to intervention status? PMID- 11434077 TI - Reducing "unnecessary" antibiotic use in primary care: hard rules, soft calls. PMID- 11434078 TI - On errors. PMID- 11434079 TI - On errors. PMID- 11434080 TI - Can evidence change the rate of back surgery? A randomized trial of community based education. AB - CONTEXT: Timely adoption of clinical practice guidelines is more likely to happen when the guidelines are used in combination with adjuvant educational strategies that address social as well as rational influences. OBJECTIVE: To implement the conservative, evidence-based approach to low-back pain recommended in national guidelines, with the anticipated effect of reducing population-based rates of surgery. DESIGN: A randomized, controlled trial. SETTING: Ten communities in western Washington State with annual rates of back surgery above the 1990 national average (158 operations per 100,000 adults). PARTICIPANTS: Spine surgeons, primary care physicians, patients who were surgical candidates, and hospital administrators. INTERVENTION: The five communities randomized to the intervention group received a package of six educational activities tailored to local needs by community planning groups. Surgeon study groups, primary care continuing medical education conferences, administrative consensus processes, videodisc-aided patient decision making, surgical outcomes management, and generalist academic detailing were serially implemented over a 30-month intervention period. OUTCOME MEASURE: Quarterly observations of surgical rates. RESULTS: After implementation of the intervention, surgery rates declined in the intervention communities but increased slightly in the control communities. The net effect of the intervention is estimated to be a decline of 20.9 operations per 100,000, a relative reduction of 8.9% (P = 0.01). CONCLUSION: We were able to use scientific evidence to engender voluntary change in back pain practice patterns across entire communities. PMID- 11434081 TI - [Pulmonary embolism--at the beginning of a new age]. PMID- 11434082 TI - [Echocardiography improves the chance of conclusive results in spiral computed tomography of suspected pulmonary embolism]. AB - While highly specific for detecting thromboembolism in proximal pulmonary arteries, spiral computed tomography (spiral CT) cannot reliably exclude pulmonary embolism. Therefore "negative" spiral CT in patients with high clinical probability of acute pulmonary embolism should be considered as non-conclusive. The goal of our study was to check whether echo/Doppler could stratify patients with suspected pulmonary embolism according to the chance of obtaining a conclusive spiral CT result. Echo/Doppler recordings of 51 patients (27 F, mean aged 50 +/- 19) admitted to ICU with high probability of acute pulmonary embolism were compared with the results of spiral CT of pulmonary artery. Pulmonary embolism was revealed by spiral CT in 36 pts. who at echocardiography were found to have shorter acceleration time (AcT 86 +/- 27 vs 117 +/- 7 ms, p < 0.00001) and right ventricle enlargement (PK 30 +/- 5 vs 25 +/- 2 mm p < 0.0001). The velocity of tricuspid regurgitation (TVPG) could not be measured in 19 patients. Among patients with long AcT (> or = 120 ms) or without right ventricle enlargement 8 of 14 and 14 of 28 patients, respectively had no intrapulmonary clots at spiral CT. Echo/Doppler helps in preselection of patients in whom confirmation of PE with spiral CT is highly probable. In patients without indirect echocardiographic sings of pulmonary embolism, despite high clinical probability the chance of a conclusive spiral CT is 40-50%. This should be kept in mind when selecting the most effective diagnostic strategy in individual cases. PMID- 11434083 TI - [Prevalence of G20210A prothrombin gene mutation in Poland]. AB - The G20210A mutation of the prothrombin (PT) gene has recently been identified as a risk factor for venous thromboembolism (VTE). This mutation was shown to be present mainly among Caucasian populations, with a higher frequency in southern than in northern Europe. The aim of our study was to determine the prevalence of the PT 20210A allele in the Polish general population and in patients with a history of venous thrombosis. The patient group comprised 323 subjects with VTE before the age of 45, recurrent VTE or thrombosis in an unusual site. The control group consisted of 399 healthy individuals. Heterozygosity for the PT 20210A allele was found in 21 (6.5%) patients and 7 (1.8%) controls. In 7 (33.3%) of the 21 heterozygous patients the PT 20210A allele was associated with the factor V Leiden mutation, in 1--with the homozygous C677T mutation of the methylenetetrahydrofolate reductase (MTHFR), and in 1--with lupus anticoagulant. Our results indicate that the presence of the 20210A allele is a mild risk factor for venous thrombosis if not associated with other thrombophilic defect (odds ratio 2.2; 95% CI: 0.8-5.5). The risk is greater in double heterozygous carriers of the PT 20210A allele and factor V Leiden mutation. PMID- 11434084 TI - [Usefulness of measuring levels of d-dimer for diagnosis of hospital venous thromboembolism]. AB - D-dimer measurement with highly sensitive tests seems useful to rule out pulmonary embolism (PE) and deep vein thrombosis (DVT). However, nonspecific increase in d-dimer is common among inpatients. The aim of our study was to check: 1) whether the frequency of normal DD level in inpatients justifies its assessment as a part of diagnostic strategy for VTE, 2) whether tests that we are using are sensitive enough to exclude PE and DVT. In 27 (47%) out of 58 hospitalised patients evaluated by ultrafast ELISA (VIDAS bioMerieux), but in none of 20/58 patients with confirmed VTE, DD-level was found normal. In 35 of those patients DD was measured also with microlatex tests--Tinaquant and BC d dimer. In 14/35 patients imaging test confirmed VTE. Sensitivity, specificity and negative predictive value (NPV) respectively were following: VIDAS: 100%, 80%, 100%, Tinaquant: 100%, 48%, 100%, BC d-dimer: 29%, 90%, 70%. Our results suggest that: 1) the relatively high frequency of normal DD-level among inpatients justifies its use in diagnostic strategies involving hospitalised patients, 2) negative VIDAS test confirms its as reliability for excluding VTE while 3) high sensitivity found for Tinaquant test encourages further prospective studies, 4) sensitivity of BC d-dimer is too low to be useful for excluding VTE. PMID- 11434088 TI - [Transesophageal echocardiography--a method for prompt diagnosis of acute hemodynamically significant pulmonary embolism]. AB - Patients with acute pulmonary embolism (PE) may require prompt confirmation of PE before aggressive therapy such as embolectomy or thrombolysis. However, emergency availability of routine diagnostic tests often remains a problem. Therefore, we assessed prospectively the diagnostic value of transesophageal echocardiography (TEE), recently reported to be useful in the diagnosis of PE. TEE was performed in 76 consecutive patients (42 F, 34 M) aged 66.7 +/- 15.8 yrs with suspected acute PE with echocardiographic signs of right ventricular pressure overload, but without known coexisting cardiorespiratory diseases. Pulmonary artery thrombi (TH) were visualized at TEE in 69.7% (53) pts. PE was confirmed by high probability lung scintigraphy and/or spiral CT in all these cases. Additionally, PE was diagnosed in 11 others without TEE-reported TH. In the 12 remaining patients PE was eventually excluded (specificity 100%, sensitivity 82.8%). Therefore, TEE in the diagnosis of hemodynamically significant PE reached 100%. No clinically important adverse events were observed during TEE. CONCLUSION: Transesophageal echocardiography is reliable and safe method of prompt confirmation of hemodynamically significant acute pulmonary embolism with relatively high sensitivity in adequately preselected patients. However, due to topographic limitations negative result of TEE does not exclude PE. PMID- 11434086 TI - [Pulmonary thrombendarterectomy in treatment of patients with chronic thromboembolic pulmonary hypertension]. AB - 20 consecutive patients with thromboembolic pulmonary hypertension underwent pulmonary thromboendarterectomy. Mean pulmonary artery pressure decreased from mean 49.9 +/- 9.8 to 25.1 +/- 8.8 mm Hg (p < 0.0001), pulmonary vascular resistance decreased from mean 5.58 +/- 2.58 to 1.62 +/- 0.79 mm Hg/l*min-1 (p < 0.0001) and cardiac output increased from 3.71 +/- 1.18 to 6.92 +/- 1.64 l/min. Sixteen patients had marked clinical improvement. Two patients died in early and 2 patients in late postoperative period due to unrelieved pulmonary hypertension and postoperative complications. PMID- 11434085 TI - [Differences in hemodynamics of thromboembolic and primary pulmonary hypertension]. AB - It is not clear whether right ventricle to pulmonary artery coupling is modified by the site of vascular obstruction in patients with chronic severe pulmonary hypertension. We compared invasively (Swan Ganz) and non-invasively (echo/ /Doppler) assessed hemodynamics between two groups of patients with severe chronic thromboembolic pulmonary hypertension (CTEPH)--(n = 6; 52 +/- 24 yrs) and pulmonary arterial hypertension (PAH) (n = 5; 42 +/- 9 yrs) who had similar invasively measured right ventricular systolic pressure (CTEPH: 78 +/- 14 mm Hg; PAH: 83 +/- 17 mm Hg; p = NS), mean pulmonary arterial pressure (CTEPH: 51 +/- 10; PAH 56 +/- 11 mm Hg, p = NS) and pulmonary vascular resistance (CTEPH: 15.6 +/- 4.4 l/min; PAH: 19.2 +/- 6.1; p = ns). Patients with CTEPH have significantly shorter acceleration time corrected to ejection time (RVET): (AcT/RVET % = 24 +/- 5% vs 32 +/- 6% in PAH; p = 0.04) as well as AcT corrected by RR distance was highly significantly shorter (8 +/- 2% vs 12 +/- 2%; p = 0.006). AcT in the CTEPH group was shorter than in the PAH (60 +/- 5 vs 75 +/- 15; p = 0.047). The mid systolic deceleration was significantly more frequent in the CTEPH group than in the PAH group (88% vs 30%; p = 0.005). If the mid-systolic deceleration was present in patients with PAH, the time to mid-systolic deceleration (t-N) had tendency to be longer in CTEPH group (118 +/- 22 ms vs 150 +/- 28 ms in PAH; p = 0.09). Significant differences appeared after correction t-N to RVET (t-N/RVET % = 46 +/- 9% vs 61 +/- 4%; p = 0.027) and to RR interval (t-N/RVET % = 16 +/- 2% vs 24 +/- 1%; p = 0.002). Doppler derived RV index proposed by Tei was slightly higher in CTEPH (0.81 +/- 0.18 vs 0.65 +/- 0.32 in PAH) but not significantly. Taken together our observations indicate that dynamical coupling between RV and pulmonary arteries is more disturbed in CTEPH than in PPH despite similar levels of chronically increased PAP. PMID- 11434087 TI - [LGM inferior vena cava filters--observation of 79 patients]. AB - The aim of the study was to assess effectiveness and safety of the LGM inferior vena cava (IVC) filters in patients with venous thromboembolic disease. In the Department of Internal Medicine of Institute of Tuberculosis and Lung Diseases in Warsaw 79 LGM IVC filters have been inserted since 1993. Indications for filters placement were as follows: recurrent pulmonary embolism (pe) despite anticoagulation--17 patients (pts), severe bleeding complications of thrombolytic or anticoagulant therapy--11 pts, contraindications for thrombolytic and/or anticoagulant treatment--5 pts, massive pe--14 pts, chronic thromboembolic-major vessel pulmonary hypertension (CTEPH)--30 pts, extensive deep vein thrombosis of lower limbs or vena cava inferior in patients with urgent indications for surgery -24 pts. Each filter placement was preceded by cavography. The diagnostic procedures (mainly ultrasonography) were performed after 3-6 and 12 months in the first year then once yearly during follow-up period. Oral anticoagulants (OA) or low-molecular-weight heparins (LMWH) were instituted in the majority of patients. 58 patients are still alive, 21 patients died. Only two non-fatal episodes of recurrent pe were documented. Other complications were rare and insignificant. We have not observed excess rate of recurrent deep venous thrombosis nor thrombosis at the filter site. The LGM IVC filters are effective and safe in such selectively chosen group of patients. PMID- 11434089 TI - [Secondary anticoagulant prophylaxis with low molecular heparins or oral anticoagulants and bone mineral density]. AB - A broad spectrum of indications for low molecular weight heparin (LMWH) requires an assessment of side effects especially during prolonged administration. There are common risk factors for venous thromboembolism (VTE) and osteoporosis; heparin is "the drug of choice" for VTE treatment. The aim of our study was to assess the effect of treatment and prophylaxis with LMWH (enoxaparine sodium) and oral anticoagulant (acenocoumarol) for bone structure. Material consists of in- and outpatients. 49 densitometries were performed in 31 patients (in 15 cases double examination). We observed a decrease of bone mineral density in comparison to the initial examination in most cases: mean change of bone mass for examined areas was 3.05%. PMID- 11434090 TI - [Thrombectomy in a patient with a mobile clot in the inferior vena cava--case report]. AB - Switching from heparin to acenocoumarol was complicated by severe retroperitoneal bleeding in a 50-years old patient with massive pulmonary embolism and deep venous thrombosis. The haematomas were evacuated by surgical procedure. Planned insertion of a vena cava filter was abandoned because of a mobile clot in inferior vena cava (IVC) reaching above renal veins as evidenced by spiral computed tomography (SCT). Patient was transferred to the Surgical Department of Medical Academy in Warsaw where thrombectomy was performed. In spite of mechanical and pharmacological methods of venous thrombosis prophylactic, thrombectomy was complicated by massive proximal deep venous thrombosis of right leg and distal part of IVC. Patient was successfully treated with UFH i.v. followed by low molecular weight heparins. No bleeding complications were observed. Screening for thrombophilia and cancer were negative. This case report is an example of difficulties in clinical management in a patient who has both life-threatening thromboembolic disease and bleeding. PMID- 11434092 TI - [Use of temporal vena cava filter in a patient with a high risk of perioperative pulmonary embolism]. AB - Intraluminal caval filter placement can be applied in order to prevent pulmonary embolism. When surgery has to be performed in the patient with proximal deep venous thrombosis anticoagulant therapy should be reduced and filter placement is indicated. Temporary filter appears an interesting option, as it can be removed shortly after surgical intervention, when contraindications to anticoagulation no longer exist. However, obligatory removal of a temporary device in case of suspected filter or vena cava thrombosis emerges as a new clinical problem. We present a ease of 20 year old woman with proximal deep venous thrombosis and high risk of pulmonary embolism in whom temporary vena caval filter with heparin infusion were chosen as a method of perioperative pulmonary embolism prevention. PMID- 11434091 TI - [Recombinant hirudine in suspected heparin induced thrombocytopenia--case report of pulmonary embolism]. AB - A case of 68 years old women suffering from chronic anemia, myelodysplastic syndrome and treated with progestogen due to endometrial hypertrophy is presented. Initially she was admitted to a regional hospital because of progressive weakness and exertional dyspnea. Three months earlier she reported an episode of acute dyspnea and chest pain. On the basis of clinical symptoms and perfusion lung scintigraphy pulmonary embolism (PE) was diagnosed. Patient received i.v. heparin which was changed to s.c. nadroparine subcutaneously. Platelet count dropped to 55,000'/ml on fifth day of treatment from initial level of about 200,000'/ml. Heparin induced thrombocytopenia was diagnosed, heparin was stopped and ticlopidine was recommended. After 3 weeks symptoms suggesting recurrent PE were observed. The patient was transferred to National Tuberculosis and Lung Diseases Research Institute. Recombinant hirudine (Refludan) was administrated (bolus 0.4 mg/kg and initial dose of infusion 0.1 mg/kg/h) overlapping with acenocoumarol from second day. Dose of r-hirudine was adjusted to achieve APTT prolongation 1.5 to 2.5 times of mid-normal range. During treatment with r-hirudine no bleeding and new thromboembolic complications occurred. Platelets count remained within normal range. After 14 days clinical improvement was observed, though symptoms of right ventricular overload and hypoxemia were still present after 6 months of treatment with oral anticoagulants suggesting chronic thromboembolic pulmonary hypertension. PMID- 11434093 TI - [Inherited thrombophilia: pathogenesis, clinical and treatment aspects]. PMID- 11434096 TI - Heavy hitters: how the Fortune 500 pick health plans. PMID- 11434095 TI - How health promotion outweighed STD costs. PMID- 11434094 TI - Can your health plan handle depression? PMID- 11434097 TI - Screening is key to preventing colorectal cancer. PMID- 11434098 TI - Work/life value can be measured. PMID- 11434099 TI - Bringing health care inside. PMID- 11434100 TI - Data watch. The scoop on Rx drug spending. PMID- 11434102 TI - The sugar model: catalytic flow reactor dynamics of pyruvaldehyde synthesis from triose catalyzed by poly-l-lysine contained in a dialyzer. AB - The formation of pyruvaldehyde from triose sugars was catalyzed by poly-l-lysine contained in a small dialyzer with a 100 molecular weight cut off (100 MWCO) suspended in a much larger triose substrate reservoir at pH 5.5 and 40 degrees C. The polylysine confined in the dialyzer functioned as a catalytic flow reactor that constantly brought in triose from the substrate reservoir by diffusion to offset the drop in triose concentration within the reactor caused by its conversion to pyruvaldehyde. The catalytic polylysine solution (400 mM, 0.35 mL) within the dialyzer generated pyruvaldehyde with a synthetic intensity (rate/volume) that was 3400 times greater than that of the triose substrate solution (12 mM, 120 mL) outside the dialyzer. Under the given conditions the final yield of pyruvaldehyde was greater than twice the weight of the polylysine catalyst. During the reaction the polylysine catalyst was poisoned presumably by reaction of its amino groups with aldehyde reactants and products. Similar results were obtained using a dialyzer with a 500 MWCO. The dialyzer method of catalyst containment was selected because it provides a simple and easily manipulated experimental system for studying the dynamics and evolutionary development of confined autocatalytic processes related to the origin of life under anaerobic conditions. PMID- 11434101 TI - Concentration by evaporation and the prebiotic synthesis of cytosine. AB - The efficient prebiotic synthesis of cytosine from urea and cyanoacetaldehyde (CA) has recently been claimed to be invalid on the basis of possible side reactions of the starting materials and the inapplicability of prebiotic syntheses using drying beach conditions. We therefore have investigated the synthesis of cytosine and uracil from urea and cyanoacetaldehyde at 100 degrees C under dry-down conditions, and in solution at 4 degrees C and -20 degrees C. We find that cytosine is produced from the low temperature experiments more efficiently than calculated from the Arrhenius extrapolation from higher temperatures, i.e., 60-120 degrees C. In addition, we find that CA dimer is as efficient as the monomer in cytosine synthesis. We also studied whether evaporating very dilute solutions of nonvolatile organic compounds will concentrate according to theory. Solutions as dilute as 10(-4) M concentrate from pure water approximately according to theory. Similar solutions in 0.5 M NaCl have less than theoretical concentrations due to absorption, but concentrations near dryness were very high. PMID- 11434103 TI - The specificity of peptide chain extension by N-carboxyanhydrides. AB - We have used amino acids activated by carbonyldiimidazole to study the enantiospecificity of peptide elongation in aqueous solution. Peptide 'primers' Glu10 and Ala3Glu10 were elongated with the enantiomers of arginine, glutamic acid, asparagine, phenylalanine, serine and valine. The homochiral addition was always the more efficient reaction; the enantiospecificity was large in some cases but very small in others. In every case Ala3Glu10 was elongated more efficiently than Glu10. PMID- 11434104 TI - Homochirality of biomolecules: counter-arguments against critical notes. AB - Three recently published critical papers by Bonner and coworkers on the extraterrestrial origin of the homochirality of biomolecules and the amplification of tiny enantiomeric excess are discussed. The presented arguments show the difficulties involved in circularly polarized u.v. and visible synchrotron radiation from neutron stars, in their ratio to non circularly polarized light and in racemization of the products. Attention is called upon another mechanism for production of extraterrestrial handedness based on Salam's condensation theory and on the recent experimental demonstration of the enantioselective magnetochiral photochemistry. Arguments, as well as experiments, point out that tiny enantiomeric excesses might be amplified via the Yamagata accumulation principle questioned by Bonner. PMID- 11434105 TI - On the relative content of G,C bases in codons of amino acids corresponding to class I and II aminoacyl-tRNA synthetases. AB - We have analyzed the relative G,C content from protein coding regions of 530 organisms and found that the ratio of the G,C content of the codons of the amino acids corresponding to Class II and Class I aminoacyl-tRNA synthetases decreases in a statistically significant way from prokaryotes to animals. This can be interpreted assuming that an initial asymmetry between the G,C content of codons of Class I and II amino acids existed and has decreased in the course of evolution. PMID- 11434106 TI - A coupled ecosystem-climate model for predicting the methane concentration in the Archean atmosphere. AB - A simple coupled ecosystem-climate model is described that can predict levels of atmospheric CH4, CO2, and H2 during the Late Archean, given observed constraints on Earth's surface temperature. We find that methanogenic bacteria should have converted most of the available atmospheric H2 into CH4, and that CH4 may have been equal in importance to CO2 as a greenhouse gas. Photolysis of this CH4 may have produced a hydrocarbon smog layer that would have shielded the surface from solar UV radiation. Methanotrophic bacteria would have consumed some of the atmospheric CH4, but they would have been incapable of reducing CH4 to modern levels. The rise of O2 around 2.3 Ga would have drastically reduced the atmospheric CH4 concentration and may thereby have triggered the Huronian glaciation. PMID- 11434109 TI - Structural infestations by imported fire ants: a survey with health implications. AB - This study provides the first baseline estimate of the impact of indoor IFA infestations in all 46 counties of our state. Clinicians need to be alert to the threat of unprovoked stings and possible anaphylaxis among disabled or confined patients in a variety of indoor settings. PMID- 11434107 TI - Survival of microorganisms under the extreme conditions of the Atacama Desert. AB - Spores of Bacillus subtilis, conidia of Aspergillus niger, versicolor and ochraceus and cells of Deinococcus radiodurans have been exposed in the dark at two locations (at about 23 degrees S and 24 degrees S) in the Atacama Desert for up to 15 months. B. subtilis spores (survival approximately 15%) and A. niger conidia (survival approximately 30%) outlived the other species. The survival of the conidia and spores species was only slightly poorer than that of the corresponding laboratory controls. However, the Deinococcus radiodurans cells did not survive the desert exposure, because they are readily inactivated at relative humidities between 40 and 80% which typically occur during desert nights. Cellular monolayers of the dry spores and conidia have in addition been exposed to the full sun light for up to several hours. The solar fluences causing 63% loss in viability (F37-values) have been determined. These F37-values are compared with those determined at other global locations such as Punta Arenas (53 degrees S), Key Largo (25 degrees N) or Mainz (50 degrees N) during the same season. The solar UVB radiation kills even the most resistant microorganisms within a few hours due to DNA damages. The data are also discussed with respect to possible similarities between the climatic conditions of the recent Atacama Desert and the deserts of early Mars. PMID- 11434108 TI - Physical activity and health among older South Carolinians. PMID- 11434110 TI - Spontaneous renal artery dissection in a healthy man. AB - We have presented a unique case of isolated renal artery dissection in an otherwise healthy young man, whose diagnosis was demonstrated by renal angiography. He was anticoagulated with warfarin for one year with resolution of the false channel in his renal artery as demonstrated by magnetic resonance angiography. Duplex ultrasonography of his renal artery was important in monitoring his renal artery flow velocities. PMID- 11434112 TI - Physical diagnosis and the technological imperative. PMID- 11434111 TI - Graduate survey of the South Carolina Area Health Education Consortium family practice residency programs. AB - The results of this study demonstrate several interesting characteristics of the graduates of the SC AHEC associated family medicine residency programs: 45 percent practice in South Carolina, 63 percent live further than 120 miles from their residency program, 96 percent are satisfied with their specialty choice, and 56 percent are involved in teaching medical students and residents. Furthermore, these graduates have the following tendencies: to practice in the traditional solo or group practice; to practice in a suburban community, town or rural community and a setting size less than a population of 100,000 persons; to care for the aging adult and geriatric population; to provide nursing home care; and to utilize house calls to provide patient care). As the current health care system continues to be redesigned, this information will be essential for assessment and planning purposes. PMID- 11434115 TI - [The inferior rectal nerve. Anatomical bases of perineal pain]. AB - MATERIAL: 4 corpses (1 fresh, 3 formoled) without preliminary injection; age: 84 to 90. METHOD: a front access enabled us to show the origin of the inferior rectal nerve from the pudendal plexus, its course across the Alcook canal and the ischiorectal fossa. Then a posterior transgluteal access showed the short intragluteal course across the "biligamentary tunnel" (between sacrospinal and sacrotuberal ligaments) of nerves and vessels related to the anus, in particular the inconstant Morestin's nerve. RESULTS AND CONCLUSION: the compression of nerves causing perineal pain can occur at different places. Depending on the localisation of the origin and the course of the inferior rectal nerve (which change) in relation to the place of that compression, and also the existence or not of Morestin's nerve, the changing topography of these pains can be explained. PMID- 11434116 TI - [Appearance of the branchial system in cervicofacial embryology]. AB - The aortic arch, vascular component of the branchial segment, is considered as its determining element. This is only justified in the conception of a branchial primordium, although this has now been discarded. The direct objections against this theory of a vascular preeminence are presented. Therefore, a revision of cervicofacial morphogenesis (formations cranial to the mandibular arch) is mandatory, based on somitomeres, neural systematization, evidence obtained from heterograft experiments and genetics, allowing to recognize in the "branchial" organization its proper somatic participation. The repartition of elements deriving from the para-axial in the "branchial" segmentation suggests that both types of segmentation represent two coordinate expressions of a single process. PMID- 11434113 TI - [Role of the lymphatics of the diaphragm in the absorption of intraperitoneal liquids]. AB - The diaphragm is the major site of the lymphatic absorption of the intra peritoneal liquids. Known since the middle of the last century, this lymphatic network is at present studied under transmission electron microscopy. The stomata which are intercellular sluices between adjacent mesothelial cells, are the entry of the diaphragmatic network. These stomata open into the lacunae which are dilatations the diaphragmatic subserous lymphatic network. The architecture of these structures explains their one-way character from the abdomen to the thorax and the role of the respiratory movements. This network collects the fluids into the trans diaphragmatic lymphatics. Pleural effusion appears when the quantity of liquids in the diaphragmatic lymphatic network exceeds the capacities of drainage of the lymphatic efferents, thus explaining the reactional pleural effusion caused by underdiaphragmatic inflammatory processes. PMID- 11434114 TI - A bilateral location anomaly of the parotid gland (case report). AB - During an anatomic dissection of the parotid region inferiorly located parotid glands were noted on both sides. Because within the glands lies the external carotid artery, retromandibular vein and the facial nerve, surgeons should have detailed knowledge of the anatomy of this region and possible variations to avoid risk of damaging, especially the facial nerve. Also the unusual location of the gland may be confused with pseudomass during CT or MRI. PMID- 11434117 TI - [The fibrous frame of the rotator cuff. Contributions of an in vivo three dimensional study of the tendons of the rotator cuff]. AB - The presence of a fibrous frame within the rotator cuff has been recently emphasized. This frame may be of interest in understanding of shoulder physiology and in attempts to improve shoulders disorders treatments. We report an original method to study the fibrous frame. MRI were done in healthy volunteers. 3D reconstruction were obtained and provided a 3D image of the fibrous frame. This technic should allow to calculate, for instance, the angles between the cuff muscles and the upper end of the humerus. PMID- 11434118 TI - [Treatment of pulmonary non-small-cell cancer--stagnation or progress?]. PMID- 11434119 TI - [Mononucleosis--diagnosis and treatment]. PMID- 11434120 TI - [Phantom pain after amputation of extremities]. AB - Phantom pain is experienced by about 70% of patients following limb amputation. In most patients, both the frequency and the intensity of pain attacks diminish with time, but severe pain persists in about 5-10%. The mechanisms underlying pain in amputees are not fully understood, but factors in both the peripheral and central nervous system play a role. Pain before the amputation seems to increase the risk of phantom pain, but the relation is not simple. Treatment of phantom pain is not successful; a recent study on prevention of the pain with a preoperative epidural blockade showed negative results. The future may reveal new specific drugs for treatment. PMID- 11434121 TI - [How many prescription drugs does a general practitioner handle? A prescription database study]. AB - INTRODUCTION: General practitioners must know of treatments for a broad range of diseases, but it may be difficult to keep abreast of the effects and adverse reactions of a large spectrum of drugs, and a good quality of prescribing implies the use of a limited number of analogue products. The aims were to investigate the number of different drugs prescribed per practice and to analyse the influence of practice characteristics on this number. MATERIAL AND METHODS: The study comprised 173 general practices (99 single and 74 group practices in the County of Funen, Denmark. All prescription registered on the Odense Pharmaco epidemiological Database (OPED) were analysed. The main outcome measure was the number of different drugs prescribed per dispensing unit. RESULTS: The number of different drugs prescribed per dispensing unit varied almost fourfold (range: 102 to 381) and four practice characteristics were able to predict 74% of this variation. Practices with several doctors, a high number of patients listed per doctor, a high percentage of elderly patients, and a heavy workload showed the highest number of different drugs prescribed. CONCLUSION: There is almost a fourfold variation in the number of different drugs used in general practice, and three-quarters of this variation is associated with factors related to the practice. PMID- 11434122 TI - [Study of selected methods of self-assessment of health, quality of life and satisfaction with treatment. Use among patients four weeks after discharge from a psychiatric ward]. AB - INTRODUCTION: The purpose of the investigation was to test selected methods for self-evaluation of health, the quality of life, and satisfaction with treatment. As part of this, to test their fitness as quality estimates in a psychiatric ward. MATERIAL AND METHODS: Four weeks after discharge questionnaires were sent to all patients and they were asked to participate. The methods used were SF-36 (short-form-36 questions), SIP (sickness impact profile), WHO-5, and a scale measuring patient satisfaction with treatment and care. RESULTS: The response rate was almost 50%. Seventy-five per cent were feeling better or the same as they did one year before admission. Compared with average values in a healthy Danish population, the psychic health of the patients was much lower, as evaluated by the SF-36 and WHO-5. The functional level measured by the SIP was reduced 12-32% from the optimum. About 75% of the patients rated the treatment and care positively. DISCUSSION: The applied methods of self-evaluation of health, quality of life, and satisfaction with treatment were found to be appropriate. For one year, it was possible to obtain and register important data from participating patients without using too many resources. Nevertheless, only 50% answered the questionnaire. It would be desirable to simplify the necessary questionnaires, thus encouraging more patients to respond. From the agreement between the methods, the use of the SF-36, WHO-5, and the satisfaction scale would be sufficient to obtain a qualified measure of the patients' health, quality of life, and satisfaction with treatment and care. Time spent on answering could be reduced to about 20 minutes. Resources spent on registration would be so small that the methods could be used for on-line quality surveillance. PMID- 11434123 TI - [Coloanal pouch in surgery of rectal neoplasms]. AB - BACKGROUND: The aim of this study was to evaluate the rate of complications and the functional result after construction of a coloanal J pouch during low anterior resection of the rectum for cancer. DESIGN: A retrospective study of medical records and interviews with patients at follow-up. RESULTS: 32 patients were followed-up for a median of 28 months (range 12-82 months). Two patients (6%) developed anastomotic leakage, one of whom died. Two patients developed rectovaginal fistula, one of whom was given a permanent colostomy. One patient died from complications after closure of the diverting ileostomy. In two patients, local cancer recurred and four died from distant metastases. At follow up, the frequency of bowel movements was median 2/24 h (range 0.3-4). Three of 29 (10%) experienced intermittent incontinence, whereas none complained of evacuation problems. DISCUSSION: The complication rate was as expected after low anterior rectal resection, but there were unexpected severe complications after closure of the diverting stoma. The functional results were satisfactory and the construction of a colonic J pouch is recommended during low rectal resection, if the defecation pattern and the quality of life is to improve, especially within the first postoperative year. PMID- 11434124 TI - [Arm morbidity after axillary dissection for breast cancer]. AB - INTRODUCTION: Axillary dissection for breast cancer is known to leave the patient with arm morbidity. The aim of this study was to describe the occurrence of lymphoedema, subjective swelling, abnormal sensations, and pain in a consecutive group of patients operated for breast cancer. METHODS: From April to October 1992, 140 consecutive women underwent an operation for breast cancer with axillary dissection at levels one and two. Patients with bilateral breast cancer or radiotherapy towards the axilla were excluded. The surviving 124 patients were invited to an investigation on arm morbidity and a questionnaire on subjective complaints was sent to them. Medical records and pathology reports were reviewed. Ninety-five patients (77%) were examined. Oedema of the arm was assessed by circumference measurements at three points. RESULTS: Objective oedema was found in six patients (6%). Subjective swelling was reported by 24 women (25%). Of these, 14 women reported slight swelling, seven had moderate swelling, and three had severe swelling of the arm. Pain within the three months preceding the examination was reported by 31 (33%). Analgesics were taken by 13 women (14%), of whom five used analgesics daily. Abnormal sensations in the ipsilateral arm was reported by 62 (65%). Young patients in the low risk group, with removal of more than ten lymph nodes, suffered a higher degree of arm morbidity. CONCLUSIONS: The only modifiable risk factor identified is the number of lymph nodes removed in axillary dissection. Other methods of staging could probably lessen arm morbidity. PMID- 11434125 TI - ["Footballer's ankle". Results of arthroscopic treatment of anterior talocrural "impingement"]. AB - INTRODUCTION: Footballer's ankle may be caused by anterior talocrural impingement of exostosis. The aim of this study was to evaluate the outcome of arthroscopic resection of exostosis, which was performed in our department. MATERIAL AND METHODS: We reviewed 17 consecutive cases of arthroscopic surgery for the bony impingement. The patients were 17 men, with an average age of 34 years. All reported pain and limited dorsiflexion in the talocrural joint. Resection of exostosis at the distal range of tibia and partial synovectomy were performed in all cases. The postoperative range of movement was evaluated after an average of eight weeks. To complete the material, we conducted a telephone interview concerning pain and return to sports. RESULTS: Dorsiflexion had objectively improved in 59%. Seventy per cent reported less pain in the ankle after surgery, whereas 59% of the patients returned to sports, 23% had given up, because of the symptoms. CONCLUSION: In cases where anterior talocrural impingement is the cause of the symptoms, arthroscopic resection of exostosis is a safe and reasonably effective treatment. The best effect was seen on pain, whereas only half of the patients had improved movement. PMID- 11434127 TI - [Radioiodine therapy of cancer metastases?]. PMID- 11434126 TI - [Lesion of the radial profundus nerve after injection of a steroid]. AB - A case of radial epicondylitis treated with steroid injections on and distal to the radial side of the elbow, followed by radialis paresis is described. Clinical symptoms, steroid toxicity, and anatomical considerations are discussed. Diagnosis and management are briefly addressed. PMID- 11434128 TI - [Nordic article of the month]. PMID- 11434129 TI - Four strategies for recruitment and retention. AB - The combined pressures of a shrinking work force, an aging population, changing social attitudes toward work, financial constraints, and public perception of healthcare have contributed to a growing personnel problem for healthcare organizations across the country. In fact, decreasing job satisfaction among healthcare employees has them headed for the doors in search of nonhospital jobs that can offer flexible hours, more opportunities, equal or better pay, and less stress. Without sufficient numbers of personnel, healthcare organizations will not be able to meet the needs of their communities. And the need for healthcare services will continue to grow as the Baby Boomers age. PMID- 11434131 TI - Leadership conference highlights strategies for affiliated groups. PMID- 11434130 TI - Reengineering your medical staff organization: care, efficiency, and relationships. PMID- 11434132 TI - Leveraging your nursing resources. PMID- 11434133 TI - Developing the mentor/protege relationship. PMID- 11434134 TI - High touch with high tech. PMID- 11434135 TI - The Medicare prescription drug benefit. PMID- 11434136 TI - Recovering from a job loss. PMID- 11434137 TI - Cascading leadership motivates followers. PMID- 11434138 TI - Online ethics resources for healthcare executives. PMID- 11434139 TI - Clinically speaking. PMID- 11434140 TI - A model workplace: creating an effective nursing environment. PMID- 11434141 TI - Too many frequencies, too little space. PMID- 11434142 TI - Laparoscopic lashback. Safety equipment can alleviate the dangers of stray burns. PMID- 11434143 TI - Branching into finance. Duties expand beyond the realm of purchasing and supply chain management. PMID- 11434144 TI - Removing the stent stash. PMID- 11434145 TI - The burden and the heart of day. PMID- 11434146 TI - The epidemiology of work and work-related disease in Rhode Island, 1876-2001. AB - Most Rhode Island workers now work in service industries, though many workers continue to be exposed to the risks of goods production in the workplace. Most reported occupational death is related to transportation and homicide, although in Rhode Island in 1999, exposure to harmful substances or environments was a significant cause of Rhode Island's few occupationally related deaths. How much occupational and environmental exposure contributes to Rhode Island's heart disease and cancer mortality rates in unknown. Musculoskeletal injuries are the predominant form of reported non-fatal occupational injuries, but there are questions about the completeness and accuracy of data on occupational injury and death. PMID- 11434147 TI - Improving the oral health of children in Rhode Island. PMID- 11434148 TI - The quality of care in the last month of life among Rhode Island nursing home residents. PMID- 11434149 TI - Hospital preparedness for acts of bioterrorism: an assessment of emergency preparedness plans of hospitals in Rhode Island. PMID- 11434150 TI - Judicial diagnosis. Pain medication and criminal law. PMID- 11434151 TI - Images in medicine. Aortic dissection. PMID- 11434152 TI - Metastatic carcinoma mimicking primary thyroid cancer. PMID- 11434153 TI - La belle indifference: medical myth or useful marker of psychiatric disease. PMID- 11434154 TI - Advances in pharmacology. New treatments for influenza: neuraminidase inhibitors. PMID- 11434155 TI - Trends in asthma hospitalizations in Rhode Island. PMID- 11434156 TI - Cancer prevalence estimates, Rhode Island 1998. PMID- 11434157 TI - [The role of selected cell adhesion molecules in chronic renal failure patients treated conservatively]. AB - Cell adhesion molecules play a pivotal role in many biological processes. Their membrane-bound forms take part in leukocyte migration in case of inflammation and therefore determine effective immune response. Uremia is a state of immunologic imbalance. By comparison with healthy controls, levels of soluble adhesion molecules are either elevated or lowered in chronic renal failure patients. This fact proves conclusively that activation and inhibition of immunocompetent cells coexist in end stage renal disease, thus showing the complexity of immune disorders in uremia. Taking into account the competitive action between circulating adhesins and their bound counterparts, one can come to the conclusion that soluble forms' dysregulation results in the blockage of membrane-bound adhesion molecules and leads to disordered adhesion. Although the effects of change in circulating adhesins' concentrations seem evident, the reason for this change is still unclear. PMID- 11434158 TI - [Anatomic and functional disorders of the urinary tract in children with urinary tract infections]. AB - The study is based on clinical analysis of 207 case recordings of children with urinary tract infection (UTI) hospitalized in The Department of Pediatric Nephrology, University Hospital, Lublin in 1995-1999. The study comprised children aged 1-18 years, average 8.8 +/- 0.4 (77.8% girls). Recurrent UTI was observed in 76.8% of children. Acute UTI was diagnosed in 20.8% of children. In 2.4% of children UTI was detected during hospitalization because of other diseases. In all children urinary X-ray examination was performed. Anatomical abnormalities were detected in 38% of children. Vesico-ureteral reflux was diagnosed in 61 children (29.5%). In 16/25 (64%) of children with vesico-ureteral reflux renal scarring was detected by radionuclide examination. In some children urodynamic examination was performed and in all of them functional abnormalities were detected. PMID- 11434159 TI - [Peritonitis in children treated with automated peritoneal dialysis]. AB - The rate of peritonitis, its etiology and efficacy of treatment was compared between 28 children treated with APD for a total of 439 dialysis months and 24 children on CAPD treated for 328 dialysis months. The peritonitis rate was significantly lower in children on APD (1 episode per 27 months) than in children on CAPD (1 episode per 17 months) (p = 0.005). The etiology of peritonitis (68% v. 62% Gram + organisms), efficacy of treatment (100% v. 100%) and relapse rate did not differ between the machine operated and manual methods of peritoneal dialysis. PMID- 11434160 TI - [Nutritional status of children treated with continuous ambulatory peritoneal dialysis and ambulatory peritoneal dialysis]. AB - Malnutrition is a very important problem in children with ESRD on peritoneal dialysis. Examinations were conducted on 20 children (9 girls, 11 boys), 10 children were treated continuous ambulatory peritoneal dialysis (CAPD) and 10 automated peritoneal dialysis (APD). Anthropometric and serum biochemical parameters were examined. Dialysis adequacy by KT/V, SCCr, PCR were described. The most significant malnutrition was observed in youngest children with lowest BMI values and significant muscle and fat mass deficiency. Increased protein catabolic rate (PCR) in children under 6 years old was an important factor leading to malnutrition. PMID- 11434161 TI - [Evaluation of health status in children, in whom vesicoureteral reflux was confirmed during infancy]. AB - A group of 62 children (age 5-12) with vesicoureteral reflux (VR) in infancy, including those who were under either ambulatory or surgical treatment and also those whose parents didn't undergo treatment, was observed with special attention paid to the following features: evaluation of height, weight, arterial blood pressure, presence of VR, size of kidneys and renal scars in them, creatinine clearance, urine analysis and urine cultures. The analysis of studied parameters proves the necessity of constant nephrological control for many years in spite of disappearance of reflux. The reason is the high risk of recurrent urinary tract infections, arterial hypertension and worse kidney functioning. PMID- 11434162 TI - [Vesicoureteral refluxes in children during the first two years of life]. AB - Ninety-six children with vesicoureteral reflux (VUR) diagnosed by age 2 were studied. The most common indication for radiological visualization of the urinary tract was a urinary tract infection (UTI). Among 143 cases of VUR, 51% were unilateral and 49% were bilateral. Other urinary tract abnormalities coexisted with VUR in 14.6% of children. Abnormal USG result was found in 24% of children, including only 13.4% with VUR and other urinary tract abnormalities. Conservative treatment was chosen in 126 (88%) cases of VUR in 87.5% of children. Follow-up voiding cystourethrography was performed in 60.7% of children. Conservative treatment was successful in 68.6% of children. VUR disappeared in 70.9% of cases after mean follow-up of 13.7 months (including 76.2% cases of VUR grade II, 54.5% cases of VUR grade III and 50% cases of VUR grade I). Recurrent UTI and lack of improvement after conservative treatment were indications for surgical treatment in 9.8% of children. Recurrent UTI were observed in 15.6% of children (including 16.7% among those treated conservatively and 8.3% among those treated surgically) and their course was febrile in most cases. PMID- 11434163 TI - [Platelet activation in relapse idiopathic nephrotic syndrome in children]. AB - The role that platelets play in pathogenesis and thromboembolic complications of the idiopathic nephrotic syndrome (INS) in children still remains unclear. The aim of the study was to analyse of platelet activation in whole blood during first 8 weeks of ins. Study group comprised 24 children with 34 relapses of INS by ISKDC (group A). Obtained results were compared to 16 healthy children (group B). We assessed activation by the count of platelet aggregates, microparticles and surface expression of selected markers--CD62P (P-selectin), CD42b (part of von Willebrand factor receptor) at the onset INS, after 2 weeks of therapy. We found the increased counts of platelet aggregates and microparticles at the onset of INS with a systematic decrease in following 2 weeks. Furthermore, expression of CD42b was significantly lower at the beginning of therapy. There were no clear correlation between markers of activation and biochemical parameters in the study group. According to these findings we conclude that increased activation of blood platelets is an independent risk factor of thromboembolic complication in the early stages of relapse of INS. The role of platelets in pathogenesis or induction of ins relapse remains the matter for further investigation. PMID- 11434164 TI - [Peroxidation of lipids and activity of antioxidant enzymes in children with nephritic syndrome]. AB - The aim of the study was to estimate antioxidant status and reliable factors involved in antioxidant protection in children with nephrotic syndrome (NS). 37 children (20 boys, 17 girls) with nephrotic syndrome (NS), aged 2.5-17 (mean 9.5), were included into the study. Erythrocyte Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), reduced glutathione (GSH) and malonylodialdehide (MDA) was estimated using commercial kits produced by Calbiochem and Bioxytech. RESULTS: 1) Decreased mean values of following parameters in children with NS- acute phase comparing to remission phase and the control group: GSH-Px (123.88 +/ 25.02; 187.69 +/- 24.57; 266.49 +/- 21.84 mU/ml; p < 0.001), GSH: (16.6 +/- 5.99; 27.81 +/- 7.68; 43.79 +/- 7.21 mumol/L; p < 0.001). 2) Increased mean values of following parameters in adequate groups: MDA: (18.70 +/- 2.01; 14.63 +/ 1.51; 9.53 +/- 1.48 mumol, p < 0.001), SOD: 536.27 +/- 450.49; 387.16 +/- 219.49; 236.96 +/- 85.71 U/mg). IN CONCLUSION: there is increased lipid peroxidation and insufficient antioxidant defence in children with nephrotic syndrome. PMID- 11434165 TI - [Dietary antioxidants and total antioxidant status in children with nephrotic syndrome]. AB - Several studies indicate the pathophysiological importance of reactive oxygen species in patients with nephrotic syndrome (NS). The present study was designed to determine the effect of dietary antioxidants on antioxidant enzymes (SOD, GPX, GR) activity and on total antioxidant status (TAS) in children with nephrotic syndrome. 36 children with NS (19 with first episode and 17 with relapse of NS) aged 4-16 were included into the study. Total antioxidant status was estimated using two-reagent Randox Total Antioxidant Status test in plasma. All patients had normal blood pressure, normal serum creatinine level and ingested a diet appropriate for age (with individual differences). Total antioxidant status was estimated using two-reagent Randox Total Antioxidant Status test in the plasma. Glutathione peroxidase (GPX), superperoxide dysmutase (SOD) and glutathione reductase (GR) activity was using antioxidant kits (Randox). A 3-day dietary intake record was obtained from each patient and then analyzed with computer program FOOD 2.0. Laboratory investigations were performed before steroid treatment. RESULTS: 1) in children with NS TAS was significantly reduced comparing to controls (0.84 +/- 0.14, 1.21 +/- 0.62 mmol/l, p = 0.002), 2) low manganese intake was found to have negative influence on TAS (TAS = 0.38 + 14.252*Mn, p > 0.001). 3) low intake of all components of antioxidant system was found: zinc (5.6 +/- 3.5 mg/kg b.w./24 h vs 8.6 +/- 4.0 mg/kg b.w./24 h), copper (0.021 +/- 0.013 mg/kg b.w./24 h vs 0.044 +/- 0.014 mg/kg b.w./24 h), manganese (0.029 +/- 0.0021 mg/kg b.w./24 h vs. 0.067 +/- 0.023 mg/kg b.w./24 h), vitamin E (0.15 +/- 0.04 mg/kg b.w./24 h vs 0.26 +/- 0.06 mg/kg b.w./24 h) and vitamin C (0.34 +/- 0.17 mg/kg b.w./24 h vs 0.87 +/- 0.19 mg/kg b.w./24 h). CONCLUSION: In children with NS reduced antioxidant protection maybe partly associated with low intake of some vital components of the antioxidant system. PMID- 11434166 TI - [The ability to produce nitric oxide by leukocytes in whole blood of children with recurrent urinary tract infections]. AB - We examined 20 children in age from 6 till 18 years old with recurrent urinary tract infection (rUTI). The control group were 15 healthy volunteers in age from 19 till 23 years old. In all cases nitrogen oxide concentration was designated in supernatant of 48 hours leukocytes culture with using colorimetric method with Griess reagent described by Ding. The spontaneous and LPS stimulated ability to nitrogen oxide production in full blood was estimated. The nitrogen oxide index was counted from the difference of concentration of inducated and stimulated nitrogen oxide. The defective response of the leukocytes of full blood to LPS stimulation for nitrogen oxide productionin aggravation also in remission in children with rUTI and with rUTI in age till 5 years old and above 5 years old comparing to healthy ones. PMID- 11434167 TI - [Lymphocyte subpopulations of peripheral blood in children with Schonlein-Henoch purpura and IgA nephropathy]. AB - The main lymphocytes' subpopulations of peripheral blood in 21 children (mean age -9.8 +/- 3.3 y) with permanent proteinuria and haematuria, 11 with IgA nephropathy (IgA) and 10 with nephropathy in course of Schonlein-Henoch purpura (Sch-H) during intensification of symptoms were assessed on flow cytometer f. Coultier, using monoclonal antibodies. The control group consisted of 21 healthy children at the same age. The results showed decreased percentage of CD4 and increased percentage of CD8 with decreased CD4/CD8 ratio in both groups of examined children. In most cases also increased percentage of B lymphocytes (CD19) were noticed and especially in children from N IgA group increased concentration of immunoglobulin A in serum. Only in some patients increased percentage of natural cytotoxic cells NK were noticed. The results were similar either in N IgA or Sch-H groups. PMID- 11434168 TI - [Levels of selected soluble adhesion molecules in blood serum of children with chronic glomerulonephritis]. AB - Increased concentrations of circulating soluble adhesion molecules have been reported in a variety of disorders. The aim of this study was to evaluate serum sICAM-1, sVCAM-1, sE-selectin, sP-selectin concentrations in 60 children with chronic glomerulonephritis GN (49 patients with GN and syndrome nephroticum, 11- with GN only), aged 3-17 years (mean 9 years) and in 15 healthy children (control group). The histopathological diagnoses of the 27 patients were: minimal change GN--10 cases, lupus nephritis--3, mesangiocapillary GN--4, mesangial GN--4, membranous GN--1, focal glomerulosclerosis--5. It was found a significant increase of tested soluble adhesion molecules in all children with GN compared to the control, independent on the histopathological type of GN. Serum sVCAM-1 and sP-selectin concentrations were higher in children with GN and syndrome nephroticum compared with patients with GN only. The results indicate activation of platelets, leucocytes, endothelium and its damage in GN. It seems, that more advanced vascular endothelium changes and platelets activation occur in children with GN and syndrome nephroticum. PMID- 11434169 TI - [Renal gene expression of renin and transforming growth factor Beta 1 in children with glomerulonephritis]. AB - Increase of renal expression of transforming growth factor beta 1 (TGF-beta 1) gene caused by activation of the local renin-angiotensin system plays an important role in the pathogenesis of glomerulonephritis (GN). The aim of the present study was to measure the expression of renin and TGF-beta 1 genes (own modification of the RT-PCR method) in the isolated renal glomeruli or in the homogenates of renal biopsy specimens in children with various types of glomerulonephritis. The study enrolled 13 children with glomerulonephritis and 3 boys with Wilm's tumour (control group). The expression of the studied genes was presented using arbitrary units defined as multiplicity of the GAPDH gene. No significant difference was found in expression of mRNA renin in the biopsy specimens of the kidney between GN group and control group. Expression of the TGF beta 1 gene was found in biopsy specimens in all patients from the control group, and only in one GN child, the sole one who was not treated with converting-enzyme inhibitors. No transcripts of the studied genes were found in all RNA samples obtained from the renal glomeruli using the microdissection method. The RT-PCR method applied in the present study allows evaluation of renal expression of renin and TGF-beta 1 genes. The authors would like to point out that storage of biopsy specimens at -80 degrees C would not prevent the total degradation of RNA during microdissection. PMID- 11434170 TI - [Class II human leukocyte antigens in minimal change nephrotic syndrome and focal segmental glomerulosclerosis--preliminary study]. AB - The aim of research was to compare the frequencies of HLA class-II antigens between children with minimal change nephrotic syndrome (MCNS) and focal segmental glomerulosclerosis (FSGS). The two morphological courses of glomerulosclerosis were considered: FSGS as a progressive state of minimal lesions (shown by renal rebiopsy results in a given patient), and primary (idiopathic) FSGS. The study group consisted of 38 children observed at least for four years. 15 patients were diagnosed as MCNS, 17 children had minimal lesions shown in the initial biopsy findings, but later progressed to glomerulosclerosis and 6 children had idiopathic FSGS. The control group consisted of 51 healthy unrelated individuals. HLA class II antigens were typed with the microlymphocytotoxicity test and with the method of sequence specific oligonucleotide DNA probes hybridisation (PCR-SSO). In minimal change nephrotic syndrome and glomerulosclerosis which followed the formerly found minimal lesions, the significant associations with HLA-DR3, DR7, and HLA-DQ2 were found. HLA-DQ1 was significantly rare in these groups. Differently idiopathic focal segmental glomerulosclerosis associates with the presence of HLA-DR4. PMID- 11434171 TI - [Evaluation of prednisone treatment in children with IgA nephropathy and Schonlein-Henoch nephropathy according to the Waldo protocol with regard to pathomorphologic changes in renal biopsy]. AB - The aim of the study was to analyse effectiveness of long term alternate-day prednisone treatment according the protocol of Waldo (Pediatr. Nephrol. 1993, 7, 529) in children with IgA nephropathy (IgAN) and Schonlein-Henoch nephritis (HSN). Eight pts: 6 with IgAN, mean age 10.1 yrs and 2 with HSN aged 10.3 and 14.3 yrs were treated with use of alternate-day prednisone for 2.3 to 3.92 (mean 2.90 yrs). Renal biopsies were performed in all patients 2 to 72 (mean 16 mths) after onset and were graded according the classification of WHO. All pts had normal serum creatinine concentrations at presentation. 3 pts had proteinuria > 1 g/1.73 m2 per day at onset and 5 pts had macroscopic or microscopic haematuria and/or increased proteinuria and/or III grade WHO in renal biopsy. After treatment haematuria and proteinuria disappeared in 2 pts and decreased in 6 pts. Hypertension (2/8), hypercalciuria (2/8), mild weight gain (6/8) and low bone density in lumbar region (5/8) were observed during the treatment. CONCLUSIONS: 1. Long term alternate-day prednisone treatment according the protocol of Waldo allows to decrease haematuria and proteinuria in children with IgAN and HSN 2. Side effects of therapy were observed in 62.5% pts. PMID- 11434172 TI - [Results of treatment with recombinant human growth hormone in children with growth retardation in end-stage renal disease]. AB - The aim of the study was to estimate the results of recombinant human growth hormone (rhGH) treatment in children with end-stage renal disease (ESRD). 60 growth retarded children with ESRD (mean age 11.2 +/- 7.2 years) were treated with rhGH at a dose of 1-1.1 IU/kg/week. The time of observation was 24 months. Thirty children completed first year, 18--second year of treatment. The mean growth velocity prior to the treatment was 3.03 +/- 1.9, during first year of the study--7.52 +/- 2.42, during second year 6.68 +/- 2.87 cm/year. The negative correlation between growth velocity and patient's age (r = -0.39; p < 0.05) suggest the better growth results in younger children during rhGH treatment. The rhGH therapy is effective method of treatment in growth retarded children with ESRD. Side effects are rare. PMID- 11434173 TI - [Epidemiology of HBV infections and possibilities for therapeutic actions in children and adolescents with end-stage renal failure treated with dialysis]. AB - Wide spreading of prophylaxis principles of HBV infections in dialysis centers decreased the HBV infection rate in general population of dialyzed patients in Poland last years. There is neither data concerned with HBV infection epidemiology in children and adolescents, nor data about anti-viral treatment possibilities and effects in this group of dialyzed patients. The aim of the study was evaluating of HBV infection rate in patients of pediatric dialysis centers and analysis of causes of infection and efficacy of treatment. Study was based on data sent in a query-answer by 8 biggest pediatric dialysis centers, all of them treating 210 patients. HBV infection was found much more often (16.6%) than in population of all hemodialyzed patients in Poland. More than 75% non vaccinated patients was infected before dialysis therapy, remaining were infected during vaccination, before the protecting level of antibodies was gained. Big differences in HBV infection rate among centers are observed. Nowadays HCV infections (more than 40% patients infected) are a bigger issue. Only 10 patients in 5 centers had anti-viral treatment (5 with isolated HBV infection, 5 with mixed HBV/HCV infection). In 9 patients interferon-alpha and in 1 patient lamivudine was administered. Efficacy of interferon-alpha treatment was similar to the population of non-uremic children (33.3% vs. 50% of HBeAg elimination). Majority of patients quite well tolerated the drug. Only in 1 case interferon alpha treatment had to be ceased because of side effects. In a boy treated with lamivudine, after 3 months elimination of viremia and decrease of ALAT activity was observed. HBV infection in patients of pediatric dialysis centers is still a serious matter. More strict applying of vaccination against hepatitis B before dialysis treatment is needed. The possibility of HBV infections therapy is limited, mostly for economical reasons. PMID- 11434174 TI - [Evaluation of oxalate/creatinine ratio in the second morning urine sample of health school children]. AB - The evaluation of urinary oxalate excretion is one of the most important diagnostic methods in patients with urolithiasis and/or nephrocalcinosis. Since reliable 24-h urine collections are difficult to obtain in children, excretion ratios of oxalate over creatinine are increasingly being used from single urine specimens. The aim of the study was to determine the normal values of oxalate/creatinine ratios in the second morning urine sample in healthy school children. The study involved 109 children between 6 and 16 years of age. The results showed that the values of Ox/Cr ratios are decreased in older children and there was significant difference between children under and above 12 years of age (values of the 95th percentile--0.076 and 0.051 mmol/mmol respectively). The significant correlation between 24-hours urinary oxalic acid excretions and Ox/Cr ratios (r-0.756) was found. We conclude, that Ox/Cr ratio is valuable parameter for screening purposes in children. PMID- 11434175 TI - [A comparison of blood pressure load in groups of children with high and low levels of neuroticism]. AB - The significance of mental stress in the etiology and pathogenesis of essential hypertension is still a matter of disputes and controversies. The authors wondered if normotensive children prone to stress reactions show significant abnormalities in their ABPM patterns. The aim of this study was comparison of blood pressure loads in children of high and low neuroticism levels. 121 healthy children underwent the psychometric test evaluating the level of neuroticism. From this group 43 children with the lowest and 33 children with the highest levels of neuroticism were selected. The groups were comparable in respect of age, gender, overweight, obesity and low birth weight. 24-hour ambulatory blood pressure monitoring was performed. Average systolic and diastolic blood pressure loads were calculated. In neurotic boys and girls considered together and girls analysed separately both systolic and diastolic blood pressure loads were significantly higher than in their low-neurotic counterparts. In boys only systolic blood pressure load difference between high and low-neurotic group was statistically significant. Diastolic blood pressure load only tended to be higher in neurotic boys, without statistical significance. In our study neurotic children demonstrated in general significantly higher blood pressure loads than low-neurotic children. The prognostic value of this difference has not yet been determined. Perhaps further investigation would prove risk of developing hypertension in neurotic children with levels of blood pressure loads as observed in our study. If so, introducing of hypertension preventive measures could be profitable in neurotic children. PMID- 11434176 TI - [Efficacy and safety of renal biopsy in children performed with an automatic biopsy device under control of ultrasonographic imaging]. AB - The efficacy and safety of kidney biopsy in children performed with an automatic Biopty Gun device under real time ultrasound was analysed. The procedure was performed under sedation during short hospitalisation with ultrasound assessment of complications performed after 24 hours. 98% of 131 biopsies yielded representative material with an average 16 glomeruli obtained per biopsy. Symptomatic perirenal haematomas were observed in 80%, subcapsular haematomas in 3.8% and arteriovenous fistulas in 4.6% of children on post biopsy ultrasound. They resolved spontaneously in all patients. CONCLUSIONS: Real time ultrasound guidance with the use of automatic biopsy devices should become the standard practice for kidney biopsy in children due to its efficacy and safety. The procedure should be performed during short hospitalization to enable ultrasound monitoring of early complications. PMID- 11434177 TI - [Ultrasonographic examination in urinary tract infections]. AB - The authors discuss the results ultrasonographic examination in children with urinary tract infections. The study included 435 children in aged 1 months-18 years treated in our Department since 1995 until 2000 y. In 260 (59.8%) children in USG examination changes in kidney were not observed. In 175 (40.2%) children changes in kidney were found. PMID- 11434178 TI - [Reasons for urinary tract infections in children based on personal observations]. AB - The aim of this study was to analyse, basing on clinical observation, the reasons of urinary tract infection in children. The study included 1016 children with urinary tract infection /832 girls and 184 boys/ in the age between 2 weeks and 18 years old hospitalised in our Department in the period 1994-2000. The most common reasons of urinary tract infections were urinary tract abnormalities and the first among them was vesicoureteral reflux. In bacterial etiology Escherichia coli was the most common bacterial strain. PMID- 11434179 TI - [Congenital hyperammonemia in neonates treated with hemodiafiltration]. AB - Inborn defects of urea cycle often results in life-threatening hyperammonemia in neonates. The initial therapy of this disease comprises administration of benzoate sodium, arginine, lactulose, neomycin, and restrictive alimentation based on carbohydrates. Renal replacement therapy for ammonia removal should be considered for the most severe cases. We present a case report of two neonates with very rare inborn urea cycle disorders--deficiency of argininosuccinate lyase and carbamyl-phosphate synthetase, treated with spontaneous arterio-venous haemodiafiltration. PMID- 11434180 TI - [Hypothyroidism as the cause of lowered blood pressure during hemodialysis]. AB - We present a case of a 19.5 years old, hemodialyzed, girl with frequent and dramatic episodes of hypotension. She presented slightly expressed symptoms of hypothyroidism, while the serum levels of hTSH, T3, T4 and fT4 were close to the bottom border of norm. The therapy with L-thyroxin (1.75 mg/kg b. w./24h) normalized the arterial blood-pressure and led to subside coexisting symptoms of hypothyroidism. PMID- 11434181 TI - [Bilateral hydrothorax in a 13-year old girl treated with continuous ambulatory peritoneal dialysis renal replacement therapy]. AB - Bilateral hydrothorax is a very rare complication of ambulatory peritoneal dialysis in children. We present a case of a 13-year-old girl who developed bilateral acute hydrothorax during after seven months of CCPD. Peritoneal dialysis was temporarily discontinued and the child was transferred to haemodialysis. Six weeks later the patient was successfully resumed CCPD. PMID- 11434182 TI - [A nineteen year observation of a boy with branchio-oto-renal syndrome and chronic renal failure]. AB - Branchio-oto-renal syndrome is a rare autosomal dominant disorder of the first and second embryonic branchial arches and the urinary tract. It is characterized in its full expression by branchial fistulas or cysts, preauricular pits, outer, middle and inner ear defects, hearing loss, lachrymal duct stenosis, facial paralysis and mental retardation. Renal anomalies may range from mild hypoplasia to complete absence. Our report demonstrates the patient with classical BOR syndrome and severe renal insufficiency since infancy up to end stage renal failure at 18 years of age caused by bilateral renal hypoplasia. Although no definitive histological diagnosis was made, the clinical findings in our patients, like mild proteinuria, normal blood pressure, polyuria, polydypsia, hyperchloremic acidosis and typical course of renal failure support the diagnosis of oligomeganephronia in this case. PMID- 11434183 TI - [Multiple bone fractures as a rare complication in a child with chronic renal failure]. AB - Renal osteodystrophy may present with a wide spectrum of bone lesions, ranging from high bone turnover to low bone turnover. The authors present a case of multiple bone fracture in 12 year old boy with chronic renal failure. This boy was hospitalized because of retention of urea. The limping was observed by admision. In the X-rays of the long bones the multiple fracture were detected. The potential causes of these fractures and diagnostic problems were discussed. PMID- 11434184 TI - [Systemic sclerosis with renal involvement in a 15-year old girl diagnosed with lichen sclerosus]. AB - In 15-year-old girl with lichen sclerosus diagnosed 4 years ago described nephritis development in systemic sclerosis without skin manifestations of scleroderma (sclerodermia sine sclerodermia). Raynaud's phenomenon, typical microcirculatory abnormalities in capilaroscopy and positive antinuclear antibodies were observed. PMID- 11434185 TI - [Osteopenia and renal calcification in a 4.5 year old child with primary distal renal tubular acidosis treated for idiopathic renal hypercalciuria]. AB - We present a case of a 4.5 years old boy suffering from hypercalciuria caused by a not diagnosed distal renal tubular acidosis. In the age of 2.5 years, after a banal infection the routine analysis showed a presence of numerous calcium phosphates in urine. Other diagnostic procedures showed: hypercalciuria, hyperphosphaturia, rather high calcemia and high values of UCa/cr and UPO4/cr ratios. HCO3 in serum 21.2 mmol/l, pH of urine 7.0. Kidneys and urinary tract-usg normal. These results induced the family doctor to make the diagnosis: idiopathic renal hypercalciuria. He advised the therapy with hydrochlorothiazide and limitations of calcium and vitamins D3 oral supply. This decision caused an illusory positive effect: decreased the UCa/cr ratio (to 0.96 mmol/mmol) without any reduction of calcemia. After a period of 12 months the UCa/cr ratio increased up to 1.31 and calcification of renal pyramids appeared. We diagnosed the distal renal tubular acidosis (some medical informations suggested the essential distal renal tubular acidosis) and osteopenia (DXA BMD L1-L4 below -1 s.d.). The therapy with NaHCO3 (about 2.6 mmol/kg) normalized the levels of HCO3a and calcium in serum, decreased the UCa/cr ratio to values 0.09-0.16 mmol/mmol. PMID- 11434186 TI - [Patients on dialysis as renal graft recipients]. AB - Many clinical aspects associated with chronic uraemia and long-term dialysis therapy may determine both the qualification for renal transplantation and post transplant outcome. These include dialysis access, severe hyperparathyroidism, inadequate or excessive erythropoietin production, heart failure, viral hepatitis, defects of urinary tract and malnutrition. Some of them delay the qualification for transplant, the other on contrary make the need for transplantation very urgent. Selected aspects are discussed in this paper. PMID- 11434187 TI - Clinical nutrition: inadequate teaching in medical schools. AB - BACKGROUND AND AIM: The teaching of Clinical Nutrition (CN) is frequently neglected in Medical Schools, though many official institutions strongly recommend its incorporation in their curricula. This work aimed to assess CN knowledge among final-year medical students and final-year dietology diploma students. METHODS AND RESULTS: We compared the performances of final-year Medical School students who did and who did not take the CN course and final-year dietology students in a computer-based multiple choice question examination related to core CN competencies that primary-care physicians and dieticians should know and be able to put into practice. The medical students who did not take the CN course correctly answered significantly fewer questions compared with those who did and the dietology students (both p < 0.001). There was also a difference in the percentages of who passed the test: students who did not take the course: those 18%; those who did: 77%; dietology students: 76% (p < 0.001). CONCLUSIONS: There are numerous barriers to the incorporation of nutrition in Medical School curricula. The medical school students may have achieved poorer results because dietology students followed nutrition education programs later in their curriculum. Our Medical School has therefore included CN education as part of its internal medicine course since 1998. PMID- 11434188 TI - Homocysteine and cardiovascular risk in patients with diabetes mellitus. AB - BACKGROUND AND AIM: Diabetes mellitus is the most common metabolic disorder in the Western world with coronary artery disease as its leading cause of mortality. Conventional coronary risk factors do not explain the high morbidity rate and there is strong evidence linking hyperhomocystinemia and premature atherosclerosis. Therefore, we reviewed the relationship between homocysteine and diabetes mellitus. DATA SYNTHESIS: The literature dealing with diabetes mellitus, B vitamins and metformin was reviewed. Some studies show higher than normal fasting and postmethionine load levels of plasma homocysteine in diabetes patients, particularly in those with nephropathy and microalbuminutia. Metformin use and low plasma B vitamins are other potential mechanisms resulting in hyperhomocysteinemia in these patients. CONCLUSIONS: Abnormal homocysteine concentration is prevalent in subsets of diabetic patients; its relationship with excess cardiovascular morbidity is not yet clear. Consequently, large prospective studies are essential in order to follow the effects of homocysteine and its normalization on accelerated artherosclerosis in diabetes. PMID- 11434189 TI - Genetics, postprandial lipemia and obesity. AB - It has been shown that a number of candidate gene loci (APOA1, APOA4, APOC3, APOB, APOE, CETP, LPL and FABP2) explain a significant, although still rather small, proportion of interindividual variability in fasting dietary responses. Other loci code for products that play a major role in lipoprotein metabolism and are prime candidate for future studies (i.e., CYP7, ABCA1, SRBI). Some of these loci also affect postprandial lipemic responses, and the interactions between genetic and dietary factors may be further modulated by body mass index. The deleterious phenotypic associations observed with some of the genotypes of candidate genes may therefore only be expressed in subjects with a high BMI. This question requires more careful research, the findings of which may contribute towards the drafting of specific dietary recommendations for the primary and secondary prevention of coronary heart disease. PMID- 11434190 TI - Obesity and free fatty acids: double trouble. AB - The socio-economic impact of obesity, one of the most prevalent medical disorders in Western society, is mainly due to its association with a higher risk of coronary heart disease. It is likely that atherosclerosis develops against a background of obesity as a result of the insulin resistance that is invariably present in overweight and obese subjects. Fasting plasma lipids may be normal in obese subjects, but they are usually affected by postprandial hyperlipidemia, which is probably due to competition between chylomicrons and VLDL for the same metabolic pathways. The basis for the impaired clearance of atherogenic chylomicron remnants is the fact that obesity causes hepatic apo B-VLDL overproduction, and thus leads to competition with chylomicrons and their remnants at the lipolytic pathway (lipoprotein lipase and hepatic lipase) and receptor level (LDL-receptor and remnant-receptor). The overproduction of VLDL is probably caused by an enhanced hepatic flux of free fatty acids in both the postprandial (from the lipolysis of triglyceride rich particles) and postabsorptive states (from adipocytes). Weight reduction by means of life-style changes, supported by medical interventions with inhibitors of intestinal fat absorption (e.g. Orlistat) or appetite suppressants (e.g. Sibutramine), is essential in order to decrease the risk of atherosclerosis. Furthermore, improvement of risk factors can be achieved by means of fibrate treatment to modulate fasting and postprandial triglyceride levels. Treatment with cholesterol synthesis inhibitors ("statins") may reduce hepatic VLDL production and increase the clearance of atherogenic remnants by upregulating LDL-receptors, thus leading to improved fasting lipid levels and enhanced clearance of chylomicron remnants. Finally, the use of thiazolidinedione derivatives to improve insulin sensitivity may be one of the options for reducing the risk of atherosclerosis in obese subjects. PMID- 11434191 TI - The toxic trio: acute, chronic, and post-prandial hyperglycemia. PMID- 11434192 TI - Effects of statins on biomarkers of bone metabolism: a randomised trial. AB - BACKGROUND AND AIM: Recently, several studies have indicated there may be differences among statins regarding a possible association between therapy and a reduction in risk of fractures. No data from prospective randomised clinical trials designed to assess either biochemical or clinical effects on bone metabolism are yet available. We assayed levels of biochemical markers of bone formation in stored serum samples from a recently completed randomised clinical trial conducted to compare the effects of simvastatin and atorvastatin on the lipid profile of patients with hypercholesterolaemia. METHODS AND RESULTS: This 12-week, randomised, multicenter, open-label study was designed to compare the safety and lipid-lowering efficacy of simvastatin 40 mg or 80 mg with that of atorvastatin 20 mg or 40 mg in 846 hypercholesterolaemic patients. Stored serum samples from this study were analysed to compare the effects of simvastatin and atorvastatin on 2 biomarkers of bone turnover, bone-specific alkaline phosphatase (BSAP), a marker of bone formation, and C-teleopeptide of type 1 collagen (CTx), a marker of bone resorption. Treatment with simvastatin 40 and 80 mg/day, but not atorvastatin 20 and 40 mg/day, led to significant (p < 0.05) reductions in BSAP in both men (4.1-5.4% reduction) and women (4.2-7.4% reduction). In addition, there appeared to be a dose-dependent effect with greater reductions in BSAP seen with the 80 mg dose of simvastatin. Treatment with either 20 mg or 40 mg of atorvastatin had no significant effect on BSAP levels on the groups as a whole or in the gender-specific subgroups. CTx showed a small, but not statistically significant, decrease with simvastatin, again with an apparent dose-related trend. Atorvastatin treatment generally resulted in small, non significant increases in CTx. CONCLUSIONS: The present serum bone biomarker results show that treatment with simvastatin, but not atorvastatin, decreases BSAP and suggest that simvastatin may have a beneficial effect on bone turnover. PMID- 11434193 TI - Statins enhance arachidonic acid synthesis in hypercholesterolemic patients. AB - BACKGROUND AND AIM: It has been shown that statins enhance arachidonic acid biosynthesis from linoleic acid in vitro, and there is also evidence that statin treated patients have high plasma and cellular arachidonic acid levels. The aim of this study was to investigate the possible effects of statins on the desaturating steps of arachidonic acid biosynthesis in patients. METHODS AND RESULTS: Plasma polyunsaturated fatty acids (percentages and concentrations), total cholesterol, LDL-cholesterol and triglycerides were measured in three groups of hypercholesterolemic patients before and after treatment with statins or fibrates for about three years, or before and after four months on a low-fat diet. As expected, there was a significant reduction in plasma total cholesterol (-23%) and LDL-cholesterol (-27%) levels in the statin group, and a significant reduction in triglycerides (-29%) and some decrease in total cholesterol (-12%) in the fibrate group; no change was observed in the control group. The selective increase in plasma arachidonic acid levels (in terms of absolute concentration and as a percentage of total fatty acids) was of the same order as the reduction in cholesterol, and the selective increase in the product/precursor ratios for delta 5 desaturation in the statin group indicated that this key step in arachidonic acid synthesis is affected. CONCLUSIONS: Our data show that an increase in arachidonic acid synthesis due to enhanced delta 5 desaturation is a major effect of statin treatment, and has the same order of magnitude as the reduction in plasma cholesterol levels. PMID- 11434194 TI - Carotid atherosclerosis in hypercholesterolemic patients: relationship with cardiovascular events. AB - BACKGROUND AND AIM: Extracranial cerebrovascular atherosclerosis is a common feature of hypercholesterolemia and carotid lesions are good predictors of cardiovascular events in the general population. Factors associated with the carotid damage of hypercholesterolemic patients and their relationships with the occurrence of clinical events are investigated in this study. METHODS AND RESULTS: One hundred and seventeen cardiovascular event-free hypercholesterolemic subjects underwent a complete clinical examination to look for additional risk factors. A blood sample was collected for lipoprotein determination and an ultrasound high resolution B-mode imaging examination of the common carotid arteries was performed. Patients were treated according to the current guidelines during a 4-yr follow-up and all major cardiovascular events were recorded. The prevalence of subjects with increased intima-media thickness and plaque was 21.4% and 29.9% respectively, higher than in normolipidemic controls. Carotid lesions were significantly related to age, hypertension and LDL-cholesterol and HDL cholesterol levels. The relative risk of developing a major clinical event was 3.92 (95% CI 1.54-9.95, p < 0.004) among categories of carotid status. At multivariate analysis, cardiovascular events were independently related to the diagnosis of familial hypercolesterolemia (FH), baseline carotid score and mean levels of LDL-cholesterol and HDL-cholesterol during the follow-up. CONCLUSIONS: Common risk factors cooperate with plasma lipoprotein levels in increasing the frequency of carotid lesions of hypercholesterolemic patients. Since such lesions are useful predictors of clinical events, B-mode ultrasound evaluation of the carotids should be routinely included in the management of these patients. PMID- 11434195 TI - [Fatal anaphylaxis after eating wild boar meat in a patient with pork-cat syndrome]. AB - Crossed allergy between pork and cat epithelia was described by us in 1994. It is due to serum albumin. Nowadays, other bio-chemical observations allow "completion" of the syndrome by extension of the crossed reactivity between other mammal meats and other epithelia of dog and horse. The authors report an observation of the pork-cat syndrome (developing in the form of anaphylaxis, and then ending in the death of the patient), following consumption of wild boar meat. Co-factors, such as effort, taking alcohol or hormonal condition may complicate the picture to make diagnosis more difficult. PMID- 11434196 TI - Delayed-type hypersensitivity and chronic fatigue syndrome: the usefulness of assessing T-cell activation by flow cytometry--preliminary study. AB - Chronic fatigue syndrome or benign myalgic encephalomyelitis has been extensively described and investigated. Although numerous immunological abnormalities have been linked with the syndrome, none have been found to be specific. This article describes the detection of delayed-type hypersensitive responses to certain common environmental antigens in almost fifty per cent of patients with this syndrome. Such hypersensitivity can be detected by the intradermal administration of antigens derived from commensal organisms like the yeast Candida albicans albicans, and then monitoring for a systemic reaction over the following six to forty-eight hours. This approach can be consolidated by performing lymphocyte activation tests in parallel and measuring in vitro T-cell activation by Candida albicans albicans antigens by three-colour flow cytometry based on CD3, CD4 and either CD69 or CD25. Another useful parameter is the kinetics of neopterin excretion in the urine over the course of the skin test. The results showed that the intensity of the DTH response correlated with the number of T-cells activated in vitro. Various factors have been implicated in the fatigue of many patients, notably lack of sleep. However, it remains difficult to establish causality in either one direction or the other. This work is in the spirit of a multifactorial approach to the group of conditions referred to as "chronic fatigue syndrome". PMID- 11434197 TI - [The drug hypersensitivity syndrome or DRESS syndrome to phenobarbital]. AB - Drug Rash with Eosinophilia and Systemic Symptoms (DRESS syndrome) or the drug hypersensitivity syndrome is a delayed and serious skin disease. It is manifest by a severe skin reaction associated with a severe visceral attack (adenopathy, hepatitis, nephritis, interstitial pneumopathy...) and haematological anomalies (raised hypereosinophilia...). The severe visceral attack is the main cause of death, which is estimated at around 10%. The principal drugs responsible are the aromatic anti-convulsants, sulphamides and minocycline. A large number of cases have been described with phenytoin, more rarely with carbamazepine and phenobarbitone. PMID- 11434198 TI - [To be effective and not harmful, ragweed-trimming methods should be rigorous]. AB - We decided to undertake the experiments dedicated to the height of Ragweed cutting. The experiment was focused on transplanted plants of Ragweed two months earlier to study the growth of male spikes. It took place near Lyon, on the second part of July 2000. We proved with the experiment that the number of male spikes produced is, to a significant extent, much more important when the height of cutting back is 10 cm instead of 5 cm. Moreover, to inhale or to be in contact with airborne matters because of Ragweed was at the origin of allergic symptoms for the author that was not atopic. It explains that Ragweed is a pollutant agent when inhaled in large quantities. Therefore, during the eradication campaign- last two weeks of July--it results better to cut back Ragweed close as possible to the ground. From spring to mid-July it is imperative not to abrade ground whereas after this period it is not really an essential point. Moreover, Ragweed has not enough grown to be cut with a mower or an edge so cutting back deals with other types of vegetal cover. PMID- 11434199 TI - [Management of allergic children in rural and urban environments. Results of our experience in a health network setting in the High Pyrenes]. AB - The allergic children management at school seems to be more and more difficult, especially in rural areas. Far from the Universitary Hospital centres how to achieve the right diagnosis? In the country side school, inaccessible for the Emergency services, how to manage an efficient interventional group? The "Hauts Pyreneens" practitioners have decided to set up a Health care Network to answer this requirement. A managing team includes the DDASS Inspector Physician (MISP), the Academy Inspector technical advisor Physician (MRDCT) and an Allergologist Practitioner. Two commissions have been set up within the network: An administrative Commission, managed by the both MISP and MRDCT, is devoted to the maternal and elementary school on one side and to the secondary schools on the other side. It is in contact with the local collectivities, the veterinary services and the centralised kitchens. A medical Commission including the Allergologist practitioners, the school health and "PMI" physicians of the Hautes Pyrenees department works to specify a diagnosis and therapeutic consensual methodology. The necessary valuation focuses first the whole net work management as well as the Welcome Individualised Project itself, each time it is carried out at a school level. PMID- 11434201 TI - [Clinical study design]. PMID- 11434200 TI - [The power of packing]. PMID- 11434202 TI - [Fungi in feces. Disease or myth?]. PMID- 11434203 TI - [Pain and pain remedies in everyday pharmaceutics]. PMID- 11434204 TI - [Alternative forms of nutrition. Real alternatives towards the concept of "high quality nutrition" of the German Nutritional Society]. PMID- 11434205 TI - [The fetlock tunnel syndrome in horses: literature review and retrospective study]. AB - The annular ligament constriction is characterized by a disproportion between the available space and the contents within the fetlock tunnel. The main symptoms are a persisting lameness, distention of the tendon sheath, a typical "notch" when the fetlock is viewed from the side and a hyperflexion pain in the fetlock. The surgical treatment consists of the transection of the fetlock annular ligament. The conservative management can be considered as a independent therapy or as a preparation for a subsequent desmotomy. The medical records of 75 horses suffering from fetlock tunnel syndrome presented at the Veterinary Surgery Clinic of the University of Zurich were studied. 39 horses with 41 affected limbs were reexamined clinically and ultrasonographically. 70% of the surgical cases and 82% of the conservatively treated cases were judged to be sound. Altogether it can be said that the surgical case group had a success rate of 62% while the conservatively treated group showed a success rate of 58%. PMID- 11434206 TI - [Simple techniques for the internal stabilization of fractures and luxations in the sacrococcygeal region of cats and dogs]. AB - Two useful techniques for the repair of caudal sacral fractures and sacrococcygeal fractures/luxations are described. Pre- and postoperative clinical, neurological and radiological findings in 16 operated animals (13 cats and 3 dogs) were compared with findings in 17 (13 cats and 4 dogs) conservatively treated animals. In the surgically treated patients follow up findings were characterized by absence of hyperesthesia and a higher number of animals with recovered tail function. The authors suggest the use of these techniques in cases of caudal sacral fractures or sacrococcygeal fractures/luxations. PMID- 11434207 TI - [Effect of a single injection of doramectin on gastrointestinal nematode infections of sheep grazing on alpine pastures]. AB - The persistent effect of doramectin injectable against gastrointestinal nematodes was investigated in a controlled field study with 70 sheep kept on alpine pastures in Switzerland. After grazing on home pastures for four weeks 50 lambs and 20 ewes were allocated to two equal groups according to age and body weight. At turnout to alpine pasture in June doramectin (0.3 mg/kg) was administered by intramuscular injection to 25 lambs and 10 ewes (Group D), whereas control sheep (Group K) remained untreated. Animals of both groups were kept on separate pastures (altitude: 1100 m) and were rotated between three paddocks during a total grazing period of 13 weeks. After doramectin treatment faecal examinations of Group D sheep showed a marked reduction of the trichostrongyle egg output which remained close to zero for eight weeks. During this period serum pepsinogen levels did not indicate the presence of a substantial immature worm burden in the abomasal mucosa. In the last five weeks of the alpine grazing period the trichostrongyle egg counts increased markedly in the lambs of Group D. Lambs of the control group developed moderate-to-high Haemonchus-infections, and eight animals of this group had to be treated with anthelmintics. Lambs of Group D had gained significantly (P < 0.05) more weight eight weeks after turnout whereas the mean bodyweight stagnated during the last five weeks of the grazing period. The results indicate, that the single administration of 0.3 mg/kg doramectin to lambs and ewes provided good protection against severe infections with gastrointestinal nematodes for a period of approximately eight weeks. PMID- 11434208 TI - [A case of canine angiostrongylosis]. AB - A six-month old bitch presenting a sub-lingual mucocele and hematoma associated with coagulation disorders died four days after the surgical treatment of the mucocele. The necropsy revealed a canine angiostrongylosis, a disease rarely seen in Switzerland. This article summarizes the biology of Angiostrongylus vasorum and describes the lesions and symptoms caused by this cardio-pulmonary helminthosis, as well as its diagnosis and treatment. The connections between angiostrongylosis and coagulation disorders are also discussed. PMID- 11434209 TI - Evaluation of health risks caused by musk ketone. AB - Among the nitro musks, musk ketone (MK) as a synthetic compound with a typical musk odor is widely used in cosmetics. In the European Community the total amount used in fragrances has been reported to be 110 tons/a. Additionally, relevant amounts of MK are used in Indian joss sticks. As a result of its inherently low biodegradability MK has been detected in the aquatic environment (surface water, sediments, edible fish). Moreover, it has been shown that MK concentrates in human fatty tissue and breast milk, indicating that humans are constantly exposed. Several studies provided convincing evidence of lack of a genotoxic potential for MK. However, MK was identified as a strong inducer of phase I enzymes in rodents and a cogenotoxicant in vitro in human derived cells in rather low doses, suggesting that exposure to MK might increase the susceptibility to health hazards caused by carcinogens in humans. PMID- 11434210 TI - GIS-based analysis of drinking-water supply structures: a module for microbial risk assessment. AB - Water-related infections constitute an important health impact world-wide. A set of tools serving for Microbial Risk Assessment (MRA) of waterborne diseases should comprise the entire drinking-water management system and take into account the Hazard Analysis and Critical Control Point (HACCP) concept which provides specific Critical Control Points (CCPs) reflecting each step of drinking-water provision. A Geographical Information System (GIS) study concerning water-supply structure (WSS) was conducted in the Rhein-Berg District (North Rhine-Westphalia, Germany). As a result, suitability of the existing water databases HYGRIS (hydrological basis geo-information system) and TEIS (drinking-water recording and information system) for the development of a WSS-GIS module could be demonstrated. Spatial patterns within the integrated raw and drinking-water data can easily be uncovered by GIS-specific options. The application of WSS-GIS allows a rapid visualization and analysis of drinking-water supply structure and offers huge advantages concerning microbial monitoring of raw and drinking water as well as recognition and investigation of incidents and outbreaks. Increasing requests regarding health protection and health reporting, demands for a better outbreak management and water-related health impacts of global climate change are major challenges of future water management to be tackled with methods including spatial analysis. GIS is assumed to be a very useful tool to meet these requirements. PMID- 11434211 TI - Eye- and skin symptoms in German office workers--subjective perception vs. objective medical screening. AB - The question of whether perceived sensation correlates with medical tests was examined using data from the ProKlimA project (1994-1999) in which sensory perception and medical examination of Sick Building Syndrome related complaints were examined in 14 office buildings across Germany. Within the ProKlimA study, a subsample of 817 persons participated in ophthalmological examinations; dermatological tests were conducted on 925 persons using serni quantitative dermatological methods. Participants of both sub-samples responded to a questionnaire assessing sensory perception over 6 sub-scales (eyes, skin nose, mouth, throat and nervous systems) consisting of 9 to 11 items per sub-scale. Non parametric analysis revealed a significant association between medical tests of symptoms and self-reported complaints regarding skin/eyes. Break up time, a measure of tear film stability, appeared to be the best indicator for self reported eye complaints. In terms of skin symptoms, low sebaceous secretion and/or low stratum corneum hydration was associated significantly with increased skin complaints. But not all items of the sensory perception modules reflected the medical diagnoses in the same way. For instance the items "rough skin" and "dry skin" described the medical diagnosis "low sebum content" better than "irritated skin", "rash" or "spotty skin". However, sensory perception modules differed in their relationship to medical tests, so that questionnaires may be used as indicators not as substitutes for medical test data. The choice of method has to be determined by the purpose of the research project. PMID- 11434212 TI - Risk assessment of virus infections in the Oder estuary (southern Baltic) on the basis of spatial transport and virus decay simulations. AB - The large Oder (Szczecin) Lagoon (687 km2) at the German-Polish border, close to the Baltic Sea, suffers from severe eutrophication and water quality problems due to high discharge of water, nutrients and pollutants by the river Oder. Sewage treatment around the lagoon has been very much improved during the last years, but large amounts of sewage still enter the Oder river. Human pathogenic viruses generally can be expected in all surface waters that are affected by municipal sewage. There is an increasing awareness that predisposed persons can be infected by a few infective units or even a single active virus. Another new aspect is, that at least polioviruses attached to suspended particles can be infective for weeks and therefore be transported over long distances. Therefore, the highest risk of virus inputs arise from the large amounts of untreated sewage of the city of Szczecin (Poland), which are released into the river Oder and transported to the lagoon and the Baltic Sea. Summer tourism is the most important economical factor in this coastal region and further growth is expected. Human pathogenic viruses might be a serious problem for bathing water quality and sustainable summer tourism. The potential hazard of virus infections along beaches and shores of the Oder lagoon and adjacent parts of the Baltic Sea is evaluated on the basis of model simulations and laboratory results. We used two scenarios for the Older Lagoon considering free viruses and viruses attached to suspended particle matter. The spatial impact of the average virus release in the city of Szczecin during summer (bathing period) was simulated with a hydrodynamic and particle tracking model. Simulations suggest that due to fast inactivation, free viruses in the water represent a risk only in the river and near the river mouth. On the other hand, viruses attached to suspended matter can affect large areas of the eastern, Polish part of the lagoon (Grosses Haff). At the same time the accumulation of viruses on suspended particulate matter increases the likelihood of an infection after incorporation of such a particle. There is no evidence, that there is a risk of virus infections in the western part of the lagoon (Kleines Haff) or along the outer Baltic Sea coast. PMID- 11434213 TI - Biochemical and pharmacological investigations of selected cyanobacteria. AB - Cyanobacteria are a very old group of prokaryotic organisms that produce a variety of secondary metabolites with antibiotic, algicide, cytotoxic, immunosuppressive and enzyme inhibiting activities. In the last decades structures of pure compounds have been determined as phenols, peptides, alkaloids or terpenoids (Falch, 1996). Screening of lipophilic and hydrophilic extracts from cultured cyanobacteria or waterbloom material, isolated from German lakes and the Baltic sea for antiviral, antibiotic, immunomodulating and enzyme inhibiting activity in different in vitro systems revealed strains with interesting effects. These strains were cultivated in 45 litre photobioreactors to produce enough biomass for bioassay-guided isolation of the active substances. First results characterising active substances are reported. PMID- 11434214 TI - Reduction potential of microbial, odour and ammonia emissions from a pig facility by biofilters. AB - The intention of this study was the determination of the potential to reduce specific microbial bioaerosol (cultivable bacteria and fungi, total cell counts of microbes, airborne endotoxins and microbial volatile organic compounds, MVOC), odour and ammonia emissions from a pig facility by biofilters. Five identical biofilter units in half technical scale were filled with different filter materials (Biochips, coconut-peat, wood-bark, pellets + bark and compost) and connected in parallel to a piggery. The results showed obvious differences between the filter materials. Numbers of airborne cultivable bacteria were decreased by ca. 70 to 95% and the total counts of bacterial cells from ca. 25 to (>) 90%. The total amount of fungal cells was reduced by at least 60%, although the percentage of cultivable moulds in the air after passing the filters was sometimes higher than before. Airborne endotoxins and MVOC were effectively reduced by all filter materials to at least 90%. Regarding odour, the average reduction was between 40 and 83%, whereas only one of the filters proved to be capable of slightly reducing the ammonia emissions. No relationships between odour/ammonia and microbial bioaerosols with regard to the reduction efficiency of the different filter materials or the total load of the emitted air could be established. A tendency could be shown, that biofilters best capable to reduce odour emitted slightly more airborne bacteria, both cultivable and total cell counts. PMID- 11434215 TI - Review of epidemiological surveys on the prevalence of contamination of healthy cattle with Escherichia coli serogroup O157:H7. AB - This paper gathers and critically analyses the results of 26 published epidemiological surveys on the prevalence of contamination of cattle with verocytotoxin-producing E. coli (VTEC) serogroup O157:H7. These surveys have been conducted since 1986 on farms in North America (10 studies), on farms in Europe (6 studies) and at slaughterhouses prior to or just after slaughter (7 studies) or after skinning and evisceration (3 studies). The purpose of this review is to understand the first stages of the epidemiology of the infection in animals and humans (the infection process being obscure in many points) and to prepare herd based control measures to reduce the risk of O157:H7 human infection. The different statistical methods employed in these surveys, as well as the various laboratory screening methods used for detecting positive animals are presented. The observed frequencies of infected animals (animal prevalence) and herds (herd prevalence) are given as a function of localisation, year, type of industry (beef or dairy) and age. From these measured prevalence values, the risk of contamination of ground beef by E. coli O157:H7 in the first stages of the farm to-fork continuum is assessed. First, we follow the evolution of contamination frequencies from the living animal on-farm to carcasses before transformation. Then, within each set of measurements (i.e., on farm or at slaughterhouse), we identify the effects of the following factors: target population, sampling strategies and laboratory procedures. We argue that the prevalence values inferred from these measurements are very likely underestimated, due to insufficient sampling and not enough sensitive laboratory procedures (one exception being the immunomagnetic bead separation technique). No firm conclusion can be drawn as to the effects of geographical localisation and season. In those surveys, the effect of hygiene level at slaughterhouse on prevalence values is not quantitatively assessed. In addition, there is growing evidence of other sources of E. coli O157:H7 than live cattle in the farm environment, such as feed, water and water-troughs. PMID- 11434216 TI - Identification and antibiotic resistance of faecal enterococci isolated from water samples. AB - We have isolated 47 strains of presumptive faecal streptococci from different water samples. Identification was made by the method of Facklam et al. (1989). Antibiotic resistance was studied on Mueller-Hinton Agar. Twelve antibiotics were tested. High-level aminoglycoside resistance (HLAR) and resistance to glycopeptides were studied. Biochemical identification of presumptive faecal streptococci isolates gave the following results: 19 Enterococcus faecalis, 12 E. faecium, 8 E. hirae, 4 E. durans and 4 E. mundtii. E. mundtii is not included among faecal enterococci. None of the strains were resistant to glycopeptides (vancomycin and teicoplanin). Three strains of Enterococci showed HLAR. Two of them were isolated from coastal bathing waters and the other from wastewater. This suggests that water could contribute to spread of HLAR enterococci and it should be a matter of concern for public health authorities. PMID- 11434217 TI - An outbreak of viral gastroenteritis in a mother-and-child health clinic. AB - An outbreak of diarrhoeal disease in a modern mother-and-child health clinic prompted the health authorities to initiate a retrospective cohort study in order to assess the scope of the outbreak and to identify possible risk factors. The management of the clinic had been rather concerned because four similar outbreaks had occurred during the last two years. A total of 151 guests, i.e. mothers with their children, who had arrived some days before the peak of the outbreak for a three-week-stay and another 15 guests who had arrived earlier and had extended their stay were enrolled in the study which mainly focused on the possible role of treatment measures as risk factors. In addition, a total of 49 staff members were requested to provide information about symptoms, working area and attendance at work. Relevant data were available from 164 of 166 guests and 47 of 49 staff members (response rates 98.8% and 96.0%, respectively). The attack rate among guests was 44.0% (adults 27.0%, children 54.0%) and among staff 23.4%. The mean age of affected children (3.5 years) was significantly lower than that of those not affected (6.3 years). The main symptoms were diarrhoea and vomiting. The sudden start of the outbreak suggested a single source of infection which, however, remained unknown. Person-to-person transmission was supposed to be the cause of the following spread. No association between distinct treatment measures and the disease was proven by the cohort study. Norwalk-like viruses as well as astroviruses were detected by polymerase chain reaction in specimens taken from seven patients. No other enteropathogenic agents were found. Regarding the special conditions in a mother-and-child health clinic where social contacts among guests are much more frequent and intensive than among patients in a "normal" hospital, measures to prevent the spread of gastrointestinal infections should concentrate on early recognition and isolation of symptomatic individuals. Guests and staff members should be instructed to keep to the rules of personal hygiene, especially handwashing. If disinfection is required, it should be virucidal. PMID- 11434218 TI - [Oxygen therapy in the long term treatment of Meniere's disease]. AB - Endolymphatic hydrops is the hystopathological substrate characteristic of Meniere's disease. Besides the classical treatment with diuretics and/or osmotic drugs for some time, now treatment in a "pressure chamber" (OTI) has also been applied. The oxygen administered in the hyperbaric chamber can reduce the hydrops both by increasing the hydrostatic pressure and by mechanically stimulating the flow of endolymph toward the duct and endolymphatic sac. In addition, an increase is seen in the amount of O2 dissolved in the labyrinthine fluids and this contributes to recovering cell metabolism and restoring normal cochlear electrophysiological functions. Between 1992 and 1996 40 patients with monolateral Meniere's disease were studied: 15 underwent oxygen therapy at a constant pressure (2.2 ATA) (HOT), 25 with a continuous variation in pressure (from 1.7 to 2.2 ATA) (Alternobaric therapy, AOT). During the acute phase the patients underwent daily OTI treatment for 15 days in a row. The maintenance treatment called for one treatment cycle (one session a day for 5 days in a row) a month for 1 year, followed by for one treatment cycle (one session a day for 5 days in a row) every three months during the 2nd, 3rd and 4th years. The controls consisted of a group of 18 patients treated with 10% glycerol i.v. (during the acute phase) and betahystine (8 mg x 3/die) between episodes. A comparison was made of the average hearing threshold for the frequencies 500-3000 Hz (PTA), how frequently episodes of dizziness arose and extent of hearing loss in the three groups after the initial 15 days of treatment and at the end of the 4-year follow up, in compliance with the criteria laid down by the Committee on Hearing and Equilibrium in 1995. At the end of the first 15 days of treatment, there were no statistically significant differences between the three groups. At the end of the follow-up, on the other hand, hyperbaric treatment, and in particular alternobaric therapy, enabled a significant reduction in the episodes of dizziness as compared to the control group. PTA and deafness also improved significantly in the patients who had undergone hyperbaric treatment. The results of the present work show that HOT, and in particular AOT, offer a valid alternative to drugs in the treatment of Meniere's disease. PMID- 11434219 TI - [Objective evaluation of dysphonia. Possibilities and limitations]. AB - Today clinical evaluation of the presence and extent of dysphonia is still essentially based on subjective-perceptive criteria. The lack of objectivity and the one-sidedness of such an approach reinforces the desire for a quantitative, objective determination of dysphonia which could aid in making an modern, unequivocal evaluation of the presence, extent and variations in time of this disorder both upon onset and after treatment. The purpose of the present research was to determine the reliability, possibilities and limitations of an objective, parametric procedure to evaluate both the normal and pathological voice. This procedure is based on the detection of the relationship between the noise and harmonic portions in the voice (the Yumoto H/N ratio). The study involved a group of 208 subjects (148 with dysphonia and 60 normal) and the procedure was applied using computerized digital sonography and dedicated algorithms. The results of this research showed that the vocal analysis procedure developed by Yumoto is not only simple but also highly sensitive, providing an objective study of dysphonia. In fact, the comparison between the average H/N ratio recorded in our patients (1.697 dB) is significantly different from that recorded in the normal subjects (11.169 dB) (p < 0.001). This also showed a clear discrimination between the two groups with no borderline or overlapping data found. The initial evaluation does not appear to establish a relationship between abnormal H/N ratio values and the type of pathology although greater morphological destruction of the vocal cords corresponds to worse vocal quality and therefore increased noise. For the data obtained to be valid, a necessary condition is the application of a strict, precise, correct sampling and analysis method following well-defined rules. Finally, the values obtained can serve as a basis for the construction of an objective instrumental voice measurement protocol that can be used in forensic evaluation of dysphonia. Standardization of the regulations is essential to such a project. PMID- 11434220 TI - [Comparison of ultrasonography and sialography vs. magnetic resonance in the diagnosis of the primary Sjogren's syndrome]. AB - To date there is no agreement as to which imaging technique is best for the evaluation of the oral component of primary Sjogren's Syndrome (SS). The purpose of the present study has, therefore, been to determine the reliability of Magnetic Resonance (MR) in the evaluation of salivary alterations in patients with SS. The study involved 23 patients suffering from SS according to the European criteria. All the patients underwent ultrasonography and MR of the major salivary glands, parotid sialography and biopsy of the minor salivary glands. The first control group was made up of 50 healthy subjects who underwent parotid ultrasonography. The second control group comprised 23 subjects who underwent MR of the head and neck for other non parotid pathology. The ultrasonography, MR and sialography images were evaluated by a single observer during a single session and scored from 0 to 4. In the SS patients ultrasonography was abnormal in all 23 cases (100%): 3 patients showed grade 1 alterations (13%); 5 grade 2 (21.7%); 9 grade 3 (39.1%); 6 grade 4 (26.1%). In the healthy controls, grade 0 was found in 36 subjects (72%) while the remaining 14 subjects revealed grade 1 alterations (28%). Using MR imaging only one of the SS patients showed grade 0 alterations (4.3%), 7 showed grade 1 alterations (30.4%), 9 grade 2 (39.1%), 5 grade 3 (21.7%) and only 1 grade 4 (4.3%). MR imaging sensitivity was 95.8% while specificity was 100%. For ultrasonography, considering grade 1 as non pathological, we found a sensitivity of 88.4% and specificity of 100%. The MR score for SS patients was compared to that obtained with sialography and ultrasonography. There was a good correlation between MR and sialography (r = 0.528, p = 0.010) while the correlation between MR and ultrasonography was not statistically significant. This study confirms that, of the diagnostic procedures available for evaluation of salivary gland involvement in SS, the most useful initial examination is ultrasonography. When there is some doubt or there are subtleties, MR is a valid alternative to classical sialography. PMID- 11434221 TI - [Microlaryngoscopy treatment of laryngeal dysplasia with CO2 laser]. AB - Classification of laryngeal dysplasia, the most appropriate treatments and criteria for evaluation of the results is still a highly controversial issue. The objectives of the present study on the treatment of laryngeal dysplasia lesions are to: 1) evaluate the relative incidence of the various forms of dysplasia in relation to grading of the histopathological findings; 2) establish the prognosis for the various forms of dysplasia considered; 3) determine the results achieved by the author's treatment protocol according to the characteristics of the dysplasia; 4) critically evaluate the classifications of laryngeal dysplasia found in the literature in view of the results of the present study. The study involved 141 patients with vocal cord dysplasia (134 men, 7 women; mean age: 56.2 years) who had come under observation at the E.N.T. Dept. of the University of Naples "Federico II" between January 1981 and April 1998. In all cases the dysplasia was removed by CO2 laser microlaryngoscopy. Of the 141 patients 89 (63.2%) showed mild dysplasia, 14 (9.9%) moderate dysplasia, 20 (14.2%) severe dysplasia and 18 (12.7%) in situ carcinoma. The five-year survival rate showed an overall actuarial survival of 89.1% for all patients while the corrected actuarial survival was 98.5% and local disease control was 86.1%. In 17 cases (12%) the dysplasia lesion recurred, in 11 (7.8%) an infiltrating carcinoma arose. Recurrences in the dysplasia were encountered in 9% of the patients with mild lesions, 7.1% of those with moderate dysplasia, 15% of the subjects with the severe form and in 27.7% of those with in situ carcinoma. An infiltrating carcinoma arose in 5.6% of the cases of mild lesion, in 7.1% of the medium dysplasias, 5% of the severe forms and in 22.2% of those with in situ carcinoma. The recurrences and infiltrating carcinomas were successfully treated with endoscopic CO2 laser surgery. Only three cases (2.1%) required radical surgery (total laryngectomy): these were patients who had not quit smoking and who had not adhered to the planned follow-up. One of the latter patients died with widespread metastases of the laryngeal cancer. The significantly higher incidence of dysplasia recurrences (P = 0.028) and infiltrating carcinomas (P = 0.015) in those patients with in situ carcinoma shows that, when preparing classification and determining prognosis, these pathologies should be considered separately. Moreover, they require a particularly precise follow-up. In conclusion, the following observations can be drawn from the study: mild and moderate dysplasias are more frequent as compared to severe dysplasia and carcinoma in situ; carcinoma in situ should be distinguished for classification and prognosis from dysplasias, considering their different clinical course and evolution; CO2 laser surgery in these forms is a valid therapeutic approach for its precision, the achievable results, the reduced discomfort to patients and for its evident cost effectiveness. PMID- 11434222 TI - [Lymphatic metastases in the parotid region from malignant skin neoplasm of the head: considerations for surgical treatment based on personal clinical experience]. AB - The present paper considers some of the problems linked to the surgical treatment of parotid metastases. The authors base these considerations on their limited personal cases and on the most accredited literature. In the presence of cutaneous malignancies of the upper portion of the face, accurate clinical radiological evaluation must be made of the parotid and neck region. Elective parotidectomy is performed when the cutaneous carcinoma is quite extensive or located in proximity of the gland or when the case involves a melanoma. Depending on the oncological margins, a sub-total parotidectomy may be enough; however, the facial nerve must be sacrificed if there is a clinical deficit or if it is directly affected by the tumor. As the dimensions of the neoplasm increase, the feasibility of saving the nerve decreases. Prophylactic treatment of the neck is advisable in parotid metastases from cutaneous malignancies, particularly melanoma. Nevertheless, it is possible to perform a selective dissection, avoiding the V level of the neck as the incidence of its involvement is quite low. Prognosis for these tumors is extremely poor. Of the eleven patients surgically treated, only 4 (36%) survived: 2 passed away from other causes not involved in the base pathology; 2 for distant metastases but with no signs of local recurrence and 1 other for recurrence of the primary pathology in the maxillary sinus mucosa. There is no difference in the survival of the two main histological groups: 2 out of 3 of the patients with melanoma (67%) died as did 3 out of 5 (60%) of those with epidermal carcinoma. This does not, obviously, include the two patients who died from other causes. PMID- 11434223 TI - [Merkel cell carcinoma]. AB - The authors present 2 cases of Merkel cell carcinoma in the head and neck district, evaluating the characteristics and evolution. Merkel cell carcinoma is a rare, highly aggressive neuroendocrine cutaneous neoplasm which is often located in the head and neck district. This carcinoma has the same characteristics as small cell tumors and tends to significant metastatization both through the lymph nodes and through the blood. It is also prone to frequent recurrence. Today diagnosis is facilitated by electron microscopy and immunohistochemistry to search for neurofilaments and cytocheratin 20. The treatment of choice is radical surgery ensuring a tumor-free edge of at least 3 cm, associated with supplementary local radiotherapy. Recurrence is quite frequent despite the treatment strategies applied. PMID- 11434224 TI - [Laryngeal-tracheal resection in the treatment of thyroid malignancies: description of a case and review of literature]. AB - Well-differentiated thyroid carcinomas infiltrate into the respiratory tract in between 0.9 and 7% of the cases. Laryngeal-tracheal involvement--most often discovered intra-operatively--can substantially modify the surgical approach. It should be evaluated pre-operatively through laryngo-tracheoscopy with a flexible fibroscope and, in some selected cases, using CT or NMR. Thyroidectomy, associated with laryngeal-tracheal resection and termino-terminal anastomosis, ensures good oncological results without any negative effect on the incidence of post-operative complications. The present work reports the case of a 64-year-old patient who, for 5 months, had presented a swelling of increasing consistency in the left hemithyroid, fixed on the deep planes. Echography showed a 5 cm mass in the left lobe and thyroid isthmus, without suspected lymphadenopathy for metastasis. Fine needle cytology was compatible with papillary carcinoma. CT using a contrast medium revealed an infiltration into the left antero-lateral wall of the I and II tracheal ring, with submucosal extension with no significant signs of stenosis of the airway. Esophagogastroscopy was normal while tracheobroncoscopy confirmed the radiological picture and made it possible to perform a tracheal biopsy which proved positive for papillary carcinoma. The patient underwent total thyroidectomy associated with anterior compartment lymphadenectomy and crico-tracheal resection of the cricoid arch and the first 5 tracheal rings. When surgery was completed, the patient was extubated without complications. Post-operative recovery was uneventful and the patient was discharged on the 9th day after surgery. Histopathological examination confirmed the pre-operative diagnosis and made it possible to stage the lesion as pT4N1bM0 because of the presence of one prelaryngeal lymph node metastasis and another one at the right paratracheal space. Complementary I131 therapy was scheduled along with periodic laryngo-tracheoscopic controls. 16 months after surgery the anastomotic line is perfectly healed and the patient is clinically disease free. A review of the literature on the surgical treatment of well-differentiated malignant thyroid neoplasms with respiratory tract involvement through resection and laryngo-tracheal anastomosis has made it possible to trace the indications, limits and results of this type of therapeutic approach. PMID- 11434225 TI - Adjusting to disfigurement: processes involved in dealing with being visibly different. AB - This paper reviews current psychological understandings of the process of adjustment to acquired and congenital disfiguring conditions, such as burns, dermatological diseases, and cleft palate. It is primarily aimed at researchers and clinicians interested in understanding and ameliorating the psychosocial impact of such disfigurements. The literature was accessed using psychological, medical, and nursing databases. The research indicates that the experience of disfigurement is multifaceted, involving individual and societal factors. The adjustment process involves the way that disfigured people interpret their disfigurement, their self, and their encounters with others. These interpretations are likely to be influenced by the interaction between various underlying cognitive self-schemas and the social context. Efficacious interventions provide disfigured people with practical strategies to deal with social encounters and/or tackle underlying cognitive processes. However, many of the studies examined were methodologically limited or uninformed by psychological theory. Future research is needed to gain a better appreciation of the experience of living with a disfigurement and to inform the development of effective clinical interventions. In particular, there is a need for studies using longitudinal and qualitative methologies, as this would foster greater understanding of the psychological and emotional processes involved in adjusting to disfiguring conditions. PMID- 11434226 TI - Paradoxical effects of thought suppression: a meta-analysis of controlled studies. AB - Research has shown that attempts to suppress a thought can cause an increase in the frequency of the thought. These paradoxical effects of thought suppression play a key role in cognitive-behavioral models of several emotional disorders. Laboratory studies of this phenomenon, however, have yielded mixed results; and narrative summaries of the literature have not been able to draw firm conclusions about the effects of thought suppression. We used meta-analysis to quantitatively examine the magnitude of thought suppression effects across controlled studies. Moreover, we explored whether the variability in effect sizes could be explained by methodological differences within and between studies. Results indicated a small to moderate rebound effect of thought suppression that varied in magnitude depending on the nature of the target thought and the method by which thought frequency was measured. Participants with clinical diagnoses did not show larger rebound effects than nonclinical or analogue participants, however, only a few studies included clinical samples. Findings are discussed in terms of implications for the ironic process theory of thought suppression, and avenues for future research on this phenomenon. PMID- 11434227 TI - The varieties of grief experience. AB - The bereavement literature has yet to show consensus on a clear definition of normal and abnormal or complicated grief reactions. According to DSM-IV, bereavement is a stressor event that warrants a clinical diagnosis only in extreme cases when other DSM categories of psychopathology (e.g., Major Depression) are evident. In contrast, bereavement theorists have proposed a number of different types of abnormal grief reactions, including those in which grief is masked or delayed. In this article, we review empirical evidence on the longitudinal course, phenomenological features, and possible diagnostic relevance of grief reactions. This evidence was generally consistent with the DSM-IV's view of bereavement and provided little support for more complicated taxonomies. Most bereaved individuals showed moderate disruptions in functioning during the first year after a loss, while more chronic symptoms were evidenced by a relatively small minority. Further, those individuals showing chronic grief reactions can be relatively easily accommodated by existing diagnostic categories. Finally, we found no evidence to support the proposed delayed grief category. We close by suggesting directions for subsequent research. PMID- 11434228 TI - Childhood animal cruelty and interpersonal violence. AB - Conduct disorder (CD) affects 2-9% of children in this country and has been found to be relatively stable through childhood, adolescence, and into adulthood. Although many behaviors that comprise CD have been studied, there has been a lack of research on cruelty to animals. It has been suggested that animal cruelty may be exhibited by 25% of CD children and that animal abuse may be the earliest symptom evident in CD children. In addition, several studies have found a significant relationship between childhood cruelty to animals and violence toward people. Available research is reviewed in this report, including early studies on the relationship between animal cruelty and interpersonal violence, recent assessment attempts, and intervention techniques. Future research needs are also outlined and discussed. PMID- 11434229 TI - Information processing in social phobia: a critical review. AB - This review critically discusses the empirical evidence for information processing biases in social phobia. Distortions in attention, interpretation, and memory processes are analyzed as they apply to individuals with social phobia. The literature provides evidence for a specific attentional bias towards socially threatening stimuli and a specific interpretational/judgment bias towards self relevant social information. However, there is little evidence to suggest that social phobia is associated with a memory bias for socially threatening stimuli. Furthermore, the relationship between the empirical evidence from information processing studies and the cognitive model of social phobia by Clark and Wells (1995) will be discussed. PMID- 11434230 TI - Dissociative identity disorder and memory dysfunction: the current state of experimental research and its future directions. AB - Memory dysfunction is a central feature of dissociative identity disorder (DID). Following the memorial anomalies outlined by Putnam [Putnam, F. W. (1994). Dissociation and disturbances of self. In: D. Cicchetti & S. L. Toth (Eds.), Disorders and dysfunctions of the self, vol. 5 (pp. 251-265). Rochester, NY: University of Rochester Press; Putnam, F. W. (1995). Development of dissociative disorders. In: D. Chicchetti & D. J. Cohen (Eds.), Developmental psychopathology, vol. 2 (pp. 581-608). New York: Wiley], the experimental research using DID case studies and samples is reviewed. As a whole, research suggests that amnesic barriers between alter personalities are typically impervious to explicit stimuli, as well as conceptually driven implicit stimuli. Autobiographical memory deficits are also experimentally evident in DID. Although no experimental studies have addressed the issue of source amnesia or pseudomemories, there is some evidence that pseudomemories are an infrequent but real phenomenon in DID patients. Finally, potential deficits in working memory are outlined, including those relating to cognitive inhibition. Research directions are discussed throughout to further elucidate the nature of memory dysfunction in DID. PMID- 11434231 TI - Alcohol and suicidal behavior. AB - Alcohol dependence and alcohol intoxication are important risk factors for suicidal behavior. However, the mechanism for the relationship remains unclear. This review presents a conceptual framework relating alcohol to suicidal behavior. Distal risk factors create a statistical potential for suicide. Alcohol dependence, as well as associated comorbid psychopathology and negative life events, act as distal risk factors for suicidal behavior. Proximal risk factors determine the timing of suicidal behavior by translating the statistical potential of distal risk factors into action. The acute effects of alcohol intoxication act as important proximal risk factors for suicidal behavior among the alcoholic and nonalcoholic alike. Mechanisms responsible for alcohol's ability to increase the proximal risk for suicidal behavior include alcohol's ability to: (1) increase psychological distress, (2) increase aggressiveness, (3) propel suicidal ideation into action through suicide-specific alcohol expectancies, and (4) constrict cognition which impairs the generation and implementation of alternative coping strategies. Moreover, the proximal risk factors associated with acute intoxication are consistent with Baumeister's (1990) escape theory of suicide. Suggestions for additional research are discussed, including the possibility that a nonlinear cusp catastrophe model characterizes the relationship between alcohol intoxication and suicidal behavior. PMID- 11434232 TI - Getting the scale right: a comparison of analytical methods for vulnerability assessment and household-level targeting. AB - This paper introduces broad concepts of vulnerability, food security and famine. It argues that the concepts and theories driving development and implementation of vulnerability assessment tools are related to their utility. The review concludes that socio-geographic scale is a key issue, and challenge. It analyses three vulnerability assessment (VA) methods, using Ethiopia as a case study. Facing the challenges of vulnerability assessment and early warning requires providing accurate information at the required scale, useful for multiple decision-makers within realistic institutional capacities. PMID- 11434233 TI - Review of policies and guidelines on infant feeding in emergencies: common ground and gaps. AB - Recent crises in regions where exclusive breastfeeding is not the norm have highlighted the importance of effective policies and guidelines on infant feeding in emergencies. In 1993, UNICEF compiled a collection of policy and guideline documents relating to the feeding of infants in emergency situations. In June 2000 Save the Children, UK, UNICEF and the Institute of Child Health undertook a review of those documents, updating the list and identifying the common ground that exists among the different policies. The review also analysed the consistency of the policy framework, and highlighted important areas where guidelines are missing or unclear. This article is an attempt to share more widely the main issues arising from this review. The key conclusions were that, in general, there is consensus on what constitutes best practice in infant feeding, however, the lack of clarity in the respective responsibilities of key UN agencies (in particular UNICEF, UNHCR and WFP) over issues relating to co ordination of activities which affect infant-feeding interventions constrains the implementation of systems to support best practice. Furthermore, the weak evidence base on effective and appropriate intervention strategies for supporting optimal infant feeding in emergencies means that there is poor understanding of the practical tasks needed to support mothers and minimise infant morbidity and mortality. We, therefore, have two key recommendations: first that the operational UN agencies, primarily UNICEF, examine the options for improving co ordination on a range of activities to uphold best practice of infant feeding in emergencies; second, that urgent attention be given to developing and supporting operational research on the promotion of optimal infant-feeding interventions. PMID- 11434234 TI - From policy to practice: challenges in infant feeding in emergencies during the Balkan crisis. AB - The preparation and dissemination of policy statements are necessary but insufficient to prevent the inappropriate use of infant-feeding products in emergencies. The widespread failure of humanitarian agencies operating in the Balkan crisis to act in accordance with international policies and recommendations provides a recent example of the failure to translate infant feeding policies into practice. This article explores the underlying reasons behind the failures which include: (1) the weak institutionalisation of policies; (2) the massive quantities of unsolicited donations of infant-feeding products: (3) the absence of monitoring systems; (4) inadequate co-ordination mechanisms; (5) the high costs of correcting mistakes; and (6) the cumulative effects of poor practice. Efforts to uphold best practice during the crisis are also documented. Finally, the article identifies actions that could be undertaken in advance of and during future emergencies to enhance the application of infant feeding policies in emergencies. PMID- 11434235 TI - Rapid assessment of population size by area sampling in disaster situations. AB - In the initial phase of a complex emergency, an immediate population size assessment method, based on area sampling, is vital to provide relief workers with a rapid population estimate in refugee camps. In the past decade, the method has been progressively improved; six examples are presented in this paper and questions raised about its statistical validity as well as important issues for further research. There are two stages. The first is to map the camp by registering all of its co-ordinates. In the second stage, the total camp population is estimated by counting the population living in a limited number of square blocks of known surface area, and by extrapolating average population calculated per block to the total camp surface. In six camps selected in Asia and Africa, between 1992 and 1994, population figures were estimated within one to two days. After measuring all external limits, surfaces were calculated and ranged between 121,300 and 2,770,000 square metres. In five camps, the mean average population per square was obtained using blocks 25 by 25 meters (625 m2), and for another camp with blocks 100 by 100 m2. In three camps, different population density zones were defined. Total camp populations obtained were 16,800 to 113,600. Although this method is a valuable public health tool in emergency situations, it has several limitations. Issues related to population density and number and size of blocks to be selected require further research for the method to be better validated. PMID- 11434236 TI - The accuracy of mortality reporting in displaced persons camps during the post emergency phase. AB - For humanitarian organisations, accurate data are essential to identify emerging health problems and determine programme needs. We visited 45 post-emergency phase displaced persons camps and collected three months' mortality data which we compared with organisations' routine mortality reports. Organisations reported 612 deaths and we identified 741 deaths, for a mortality-reporting ratio, defined as the number of organisation-reported deaths divided by the number of investigator-identified deaths, of 83 per cent. For the majority of camps which under-reported deaths, mortality reporting ratios were significantly higher for women than men, and for camps with central mortality registers rather than those without. In the few camps which over-reported deaths, these occurred primarily among children younger than five years of age, probably due to the inclusion of abortions and stillbirths. Despite the overall under-reporting of deaths by humanitarian organisations, the existing health information systems appear to estimate mortality rates adequately in these post-emergency camps. However, organisations should improve the precision and completeness with which they report the characteristics of deaths in order to provide valuable data to target their programmes at the most vulnerable people. PMID- 11434237 TI - Misanthropy without borders: the international children's rights regime. AB - The issue of children's rights has become key to human rights-based international security strategies. The UN Convention on the Rights of the Child (1989) is being operationalised in complex political emergencies. Children's rights now inform humanitarian principles. Universal concern for children is viewed as transcending political and social divides and able to mobilise societies to confront social problems and prevent war. The operationalisation of child rights is accompanied by the development of psycho-social programmes to rehabilitate the child victim. Critically analysing the implications of the children's rights regime for the right to self-determination, the paper unpacks the assumptions underlying children's rights and psycho-social intervention. The paper begins by examining the conceptualisation of the rights-holding subject universalised under the UN Convention and then goes on to consider Article 39 on the right to psycho-social intervention. Equally important as the novel conceptualisation of childhood and children's rights under the international children's rights regime is the (unspoken) mistrust of adulthood and political rights that informs the imperative to institutionalise children's rights as higher law. Moreover while the rights based approach consciously sought to move away from the earlier moralising child salvation model, psycho-social rehabilitation reveals a similar preoccupation with deviancy, but conducted through the paradigm of psychological functionalism. Rather than representing a trend towards more humane international relations, the paper suggests that the elevation of children's rights is premised on a profound disenchantment with humanity. The logical implication of the international children's rights regime is to challenge both the moral and political capacity of individuals and their right to self-determination and to institutionalise a more unequal international system. PMID- 11434238 TI - Movement toward DM accreditation picks up steam. PMID- 11434239 TI - DM approach to unexplained chronic symptoms produces consistent outcomes, solid ROI. AB - High-utilizing patients are a problem for most health care organizations, particularly in cases where the root cause of a problem cannot be medically explained. An educational approach aimed at helping these individuals understand and deal with their symptoms is delivering relief to everyone involved. PMID- 11434240 TI - Mentor-driven cardiovascular risk-reduction program makes a strong case for prevention. AB - It is the biggest killer of Americans, but many experts agree that traditional medicine is too focused on fixing cardiac problems rather than preventing them. Further, there is evidence that even modest efforts which focus on behavior change can make a significant dent in utilization. See how one mentor-based program is helping providers give their patients the preventive guidance and support they need. PMID- 11434241 TI - Web-based educational effort for CHF patients boosts outcomes while cutting costs. AB - The high utilization associated with CHF can often be linked to gaps in patient knowledge and self-management skills. DM efforts aimed at showing up these elements can potentially deliver big dividends. One innovative web-based approach has not only delivered clinical and financial benefits, but it has also streamlined administrative functions and boosted productivity. PMID- 11434242 TI - [Serious eating disorders--from words to action?]. PMID- 11434243 TI - [Why vaginal hysterectomy?]. PMID- 11434244 TI - [Bulimia nervosa]. AB - Bulimia nervosa (BN) is a relatively new syndrome occurring particularly in young women. The gender ratio is about 10:1. The disease is becoming more common especially in the Scandinavian countries. The symptoms are episodes of binge eating followed by weight regulation. Generally, weight reduction is the provoking factor. The disease is estimated to occur in 3-5% of Danish women between the age of 14 and 24 years, but also in an unknown number of older women. Several factors increase the risk of BN: high body mass index, sociocultural circumstances, such as a "drive for thinness", and psychopathological conditions, for instance low self-perception, perfectionism, and a failing internal authority; presumably there is a genetic vulnerability. BN has a number of serious somatic consequences, as well as dental damage. If left untreated, the symptoms seem to persist, whereas the results of never treatment indicate a high degree of recovery. There are thus many reasons in favour of intensifying and increasing the provision of treatment options, as well as a need for prevention. PMID- 11434245 TI - [Treatment of anorexia nervosa]. AB - Anorexia nervosa is a common disorder, that is becoming an increasing burden on health care institutions. It has complicated biological and psychological symptoms that reinforce each another. The treatment is of long duration and is often difficult. Patients are ambivalent about treatment and the treating doctor must actively motivate them to continue treatment. An important part of the treatment is the re-establishment of normal body weight. Uncomplicated cases should be treated by the general practitioner. PMID- 11434246 TI - [High prevalence of eating disorders among elite athletes. Increased risk of amenorrhea and premenopausal osteoporosis]. PMID- 11434247 TI - [Mortality and causes of death in patients with eating disorders in Denmark, 1970 1993]. AB - INTRODUCTION: The aim of this study was to describe standardised mortality rates and causes of death in patients with eating disorders, who were admitted to Danish psychiatric and somatic hospitals during the period, 1970 to 1993. METHODS: By means of record-linkage, the study covered all patients admitted for and diagnosed as suffering from an eating disorder, according to the ICD-8 classification system, at any Danish psychiatric (since 1970) or somatic department (since 1977) during the period 1970-1993. The study comprised 2763 patients 237 of whom were males. The maximum follow-up time was 23 years and the mean follow-up time 10.3 years. RESULTS: The crude mortality at follow-up was 8.4%. A significant excessive mortality was demonstrated, as the standardised mortality ratio (SMR) of the total patient population was 6.69 (CI 5.68-7.83) and the highest rate ratio (RR) of 14.92 (CI 9.66-22.03) related to women aged 25-29. CONCLUSION: With its 2763 patients, this is the largest study of a population with eating disorders ever published. The study documents a significant excessive mortality among such patients in all age groups. PMID- 11434248 TI - [Vaginal hysterectomy in non-prolapsed uterus]. AB - INTRODUCTION: There is an increasing number of methods available for removing the uterus, but no multicentre, randomised, clinical trials are available to show which type of operation ensures the best outcome for the patient. The charts of 207 vaginal hysterectomies were reviewed in this descriptive study. The aim was to examine whether vaginal hysterectomy might be an alternative for a group of patients. MATERIAL AND METHODS: Only patients with a previous, vaginal birth were entered in the study (four patients had not given birth). A uterus weight of 500 600 g was the upper limit set by our group for the use of the vaginal route. Patient characteristics, data from the operations, complications, and histological examinations of the uterus were analysed. RESULTS: The mean operation time was 61 min (95% CI 58-64 min), range 25-115 min. The difference between preoperative and postoperative haemoglobin was 1.1 mmol/l (95% CI 1.0-1.2 mmol/l). The mean length of hospital stay was 2.9 days (95% CI 2.7-3.1 days). The complications were 1) febrile morbidity; 17 (8%) (95% CI 4-12%); 2) urinary tract injury, 3 (2%) (95% CI 0-3%); 3) unforeseen need for major surgery (including conversion to abdominal hysterectomy), 3 (2%) (95% CI 0-3%). DISCUSSION: In some patients, vaginal hysterectomy is a good alternative. PMID- 11434249 TI - [Low occurrence of nausea and vomiting after propofol anesthesia]. PMID- 11434250 TI - [Picture of the month: Hyperparathyroidism secondary to vitamin D deficiency]. PMID- 11434251 TI - [Restless legs syndrome. Therapy according to plan]. AB - The restless legs syndrome is characterised by an imperative desire to move lower legs because of paraesthesias or dysesthesias sometimes associated with pain witch occur at rest. The symptoms temporary relief by activity. Worsening of symptoms in the evening or at night are typical. Therefore patients with RLS often have problems with initiating and maintaining sleep. In polysomnographic investigation typical periodic limb movements in sleep (PLMS) can be found. The treatment of choice for RLS is levodopa. In case of augmentation or time shifting transfer to dopamine agonists should be considered. Alternative medications are opioids, benzodiazepines or some antiepileptic drugs (carbamazepine or gabapentin). PMID- 11434252 TI - [Diagnosis and therapy of depression. Key role for the family physician]. AB - Several epidemiological studies initiated by the World Health Organisation (WHO) have demonstrated the outstanding significance of depressive disorders in medical care and for public health aspects. Moreover it is evident that the majority of depressed patients is treated by primary care physicians, rather than by mental health specialists. Therefore, primary care physicians figure prominently in the diagnosis and treatment of depression. However, there is not always sufficient awareness of the fact that depression is among the most common and also with regard to suicidality among the most life-threatening diseases in the primary care setting. Clinical guidelines, taking into account the specific situation in primary care, should safeguard that depressive disorders are rapidly identified. As long as there are no complicating aspects, requiring the support of specialists, depression can usually be treated in primary care. Established standards for the treatment of depression would facilitate this task. Clinical guidelines for the primary care setting must be based on further clinical research. PMID- 11434253 TI - [Dementias. Diagnosis and therapy are overwhelmingly the responsibility of the family physician]. AB - The management of elderly patients suffering from dementia is a task mainly for the general practitioner, who is in a favourable position to recognize this condition at an early stage, since he knows the patient, his family and situation. The diagnosis is facilitated by recently developed screening tests. The family doctor may also be confident to make most differential diagnostic decisions. Treatment builds on basic general medical therapy. For pharmacotherapy, modern antidementia agents, antidepressants and neuroleptic agents are available. Pharmacotherapy must be supplemented by adaptation of the patient's surroundings, and counseling of relatives. However, proper treatment of dementia is cost-intensive and puts a strain on the doctor's medications budget. PMID- 11434254 TI - [Estrogens against Alzheimer disease? The female sexual hormone as a protective neurohormone]. AB - The female sex hormone estrogen has a wide range of activities besides the sex associated functions. Estrogen is a neurohormon. Its effects on structure and function of nerve cells is in the focus of research of basic neuroscience. In the brain estrogens not exclusively act via estrogen receptors but also receptor independently. Estrogen can function dependent but also independent from estrogen receptors. In humans estrogen improves cognitive functions, learning and memory. In addition, beneficial effects of a hormone replacement therapy in prevention of Alzheimer's disease have been demonstrated. But on the basis of the current knowledge, estrogen is not a drug for the treatment of an already ongoing Alzheimer's disease process. PMID- 11434255 TI - [Differential diagnosis and therapy of headache in general practice. Anamnesis is the alpha and omega]. AB - Headache is one of the most common reasons for a patient to consult the doctor. Of prime importance are the correct differential diagnosis and effective treatment. A differentiation is made between primary and secondary headache. In the international classification, the primary headache syndromes include migraine with and without an aura, tension type headache, headache associated with misuse of analgesics, cluster headache, and a number of rare forms of headache with no structural lesion. The secondary headache syndromes occur symptomatically as sequelae of underlying disease, the spectrum of causes covering more than 300 different disorders. In patients with headache of unclear genesis, careful history-taking and thorough physical examination should be followed by a further diagnostic work-up. PMID- 11434256 TI - [Stroke patients in general practice. Preventing recurrent infarct]. AB - Stroke patients have a high recurrence risk of 4-14% per year--depending on individual etiology. The best way of preventing a repeat insult and protecting the patient's remaining quality of life is to rigorously apply all available secondary prophylactic possibilities. These include measures aimed at modifying a health-endangering lifestyle, as well as medical treatment of all risk-enhancing illnesses. The present article offers an overview of the major confirmed and modifiable risk factors for stroke (arterial hypertension, smoking, atrial fibrillation, diabetes mellitus, overweight, hyper-cholesterolemia, thrombophilia, immoderate use of alcohol, lack of exercise, use of contraceptives, migraine), and outlines therapeutic strategies for secondary prevention. PMID- 11434257 TI - [Parkinson disease. Directed use of modern therapy]. AB - The approval of new antiparkinsonian drugs such as novel dopamine agonists or COMT-inhibitors during the last years has improved the therapeutic possibilities in Parkinson's disease. The choice of the different drugs depends on their particular efficiency, their adverse effect profile, the stage of the disease, the cardinal symptoms, age, and comorbidity. Therefore the physician is challenged to adjust the therapeutic management of each patient to the particular medical and social needs. This article discusses the present therapeutic strategies for the different stages of the disease detailing the profile, the main applications and the most important side effects of each drug. PMID- 11434258 TI - [Sleep disorders. Differential diagnosis and therapy in general practice]. AB - Sleep Disorders are one of the most common complaints of patients seen by general practitioners. Prevalences vary considerably--depending on the kind of research and the definition of sleep disorders--between 4 and 40%. In 2000 the first grand nationwide survey was carried out in primary care settings [1]. 12.3% of the 17,928 questioned stated sleep disorders as the main reason for their consultation, the prevalence of all kinds of sleep disorders being 29%. Like seen in other studies sleep complaints increased with age and predominated significantly in women. In one-third of the sample the differential diagnosis was primary or non-organic insomnia [1] while treatment was mainly conducted by the general practitioner. In two-thirds of the patients sleep disorders existed for more than 12 months while 41%--by their own account--took sleep agents at least once a week. This article gives an overview of the causes and treatment of sleep disorders, particularly therapy of non-organic insomnia. PMID- 11434259 TI - [Neurosurgical therapy of Parkinson disease. Deep brain stimulation]. AB - The introduction of continuous high frequency stimulation (deep brain stimulation) into functional neurosurgery has opened up new avenues in the treatment of Parkinson's disease. This new technique expands the therapeutic possibilities available to those patients in whom, over the years, the effectiveness of drug treatment has deteriorated, or severe side effects developed. In the individual case, the decision as to whether to operate is taken on the basis of interdisciplinary cooperation between the care-providing neurologist and the neurosurgeon specialized in this particular field. PMID- 11434260 TI - [Therapy of polyneuropathies. Causal and symptomatic]. AB - In the first instance, polyneuropathies are treated causally. The most common underlying cause is diabetes mellitus or alcohol abuse. In a large number of patients with polyneuropathy, however, the underlying cause cannot be definitively identified. For these--but equally for patients with etiologically clear polyneuropathy--a stock-taking of clinical symptoms should be carried out and, where indicated, symptomatic treatment initiated. In addition to medication aimed at combating pain, muscular spasm, autonomic functional disorders, and for the prevention of thrombosis, physical measures (physiotherapy, foot care, orthopedic shoes) are of primary importance. PMID- 11434261 TI - [Eletriptan shortly before drug approval. Fewer problems in migraine therapy]. PMID- 11434262 TI - [Psychiatric comorbidity in somatic disorders. Psychophytopharmaceuticals are worth a try]. AB - Somatic and psychiatric illnesses are commonly associated. Cardiovascular disorders, Parkinson's disease and stroke are especially often associated with depression as also anxiety and sleep disturbances. Psychophytopharmaceuticals, which are usually well tolerated, are highly suitable for this purpose. To treat mild to moderate episodes of depression, extracts of St. John's Wort are employed, but recently reported interactions with coumarin, theophylline and also estrogen preparations, need to be considered. Unspecific anxiety states can be treated with extracts of kava. At present, no herbal preparation can be recommended for the treatment of sleep disorders, as no unequivocal proof of efficacy in patients with such problems has yet been established. PMID- 11434263 TI - [Schizophrenia and addiction. A frequent psychiatric dual diagnosis]. AB - The dual diagnosis schizophrenia and addiction represents a combination of an endogenous psychosis and an addictive condition in the sense of abuse of or dependence on psychotropic substances. Before this diagnosis is established, schizophreniform organic psychoses must be distinguished from primary schizophrenia. Since the psychopathological differentiation is often not quite unequivocal, a search must be made for treatable underlying disorders. Here, help is afforded by the patient's history obtained on the basis of the reports of relatives and friends, physical neurologic examinations, and various laboratory, radiologic, electrophysiologic and neuropsychologic methods. In the daily clinical situation, a series of characteristic leading symptoms help clarify the differential diagnosis. The starting point for treatment is the patient's personal view of the problem, and his subjective state of health. Goals to be aimed at include integrative therapeutic approaches propagating the interaction of pharmacotherapy, psychotherapy and sociotherapy. PMID- 11434264 TI - [Established and atypical neuroleptics. What the family physician should know]. AB - Some 30-50% of schizophrenic patients fail to respond at all, or only inadequately, to the classical neuroleptics. The side effects profile is broad- in particular adverse reactions affecting the extrapyramidal motor system may be induced. In recent years, numerous, so-called atypical neuroleptic agents have been approved. These substances are characterized by improved efficacy and fewer side effects. The downside, however, is that they are considerably more expensive than the established neuroleptics. A further disadvantage is the fact that no depot preparations are as yet available. Finally, some of these substances lead to a marked increase in weight. PMID- 11434265 TI - [Extrapyramidal motor side-effects and blood profile changes. Stumbling blocks in psychiatric pharmacotherapy]. PMID- 11434266 TI - [Normal pressure hydrocephalus--from clinical picture to diagnosis. Help by early shunt placement]. AB - Normal pressure hydrocephalus (NPH), which may either be idiopathic or occur secondarily, is responsible for approximately 6% of the cases of dementional syndromes in the elderly. Although its diagnosis still remains a problem, every attempt should be made to identify patients with clinical symptoms arising from NPH, since the placement of a shunt can result in a much improved quality of life. Candidates for surgical treatment must be selected with care, since the transient peri-operative morbidity may be as high as 7%. PMID- 11434267 TI - [A classic antidepressant as an analgesic. Chronic pain under secure control]. PMID- 11434268 TI - [Opiate dependent pregnant patients and mothers. What must be regarded in drug substitution and detoxification]. AB - Substitution treatment with methadone, is a major addition to the therapeutic options available to opiate-dependent patients. In the case of opiate-dependent pregnant women and mothers with children, however, it brings other problems with it. Thus, for example, cases of methadone poisoning of children have become known. In part these are accidental, but some are a result of deliberate attempts on the part of mothers to quieten their children. The present report from an outpatient addiction service describes specific possibilities for treatment, and offers of organized help for this group of patients. PMID- 11434269 TI - [Brain protective interstitial laser thermotherapy. Therapy of brain tumors without secondary damage]. AB - The purpose of interstitial radiosurgery is to deliver a necrotizing dose of heat to an accurately defined focal area without damaging adjacent healthy brain tissue. To achieve this, heat at a temperature of 60-100 degrees C is applied via a laser fiber placed stereotactically in the center of the tumor. With the aid of thermosensitive magnetic resonance imaging (MRI), not only can the heat distribution within and around the tumor be measured during treatment, but also the extent of the lesion produced assessed. Interstitial laser thermotherapy (ILTT) performed under MRI monitoring, could become an important interdisciplinary minimally invasive treatment option for patients with brain tumors. Experimental data on the biological effects of interstitial laser therapy on normal brain tissue are not yet available, and only preliminary clinical studies investigating the effects of laser energy on brain tumors have so far been carried out. This overview presents a description of our own initial results, discusses the present state of our knowledge and current possibilities and limitations of this new treatment modality. PMID- 11434270 TI - [Antidepressive therapy. It depends on long-term maintenance]. PMID- 11434271 TI - [From taboo topic to acceptance. Senile dementia requires integrated treatment concepts]. PMID- 11434272 TI - [L-dopa-induced movement disorders in Parkinson disease. Amantadine improves dyskinesia]. PMID- 11434273 TI - [Parkinson patients. Normal life expectancy with selegiline?]. PMID- 11434274 TI - [Depressive disorders. A third have a chronic course]. PMID- 11434275 TI - [Interferon in multiple sclerosis. The earlier, the better?]. PMID- 11434276 TI - [Neuromuscular blockade with botulinum toxin. From toxin to therapeutic agent]. PMID- 11434277 TI - [Schizophrenia and diabetes alliance. No increased insulin resistance due to risperidone]. PMID- 11434278 TI - [The antidepressant nefazodone. Healthy sleep fosters compliance]. PMID- 11434279 TI - Degradation products of a phenylurea herbicide, diuron: synthesis, ecotoxicity, and biotransformation. AB - The degradation products of diuron (photoproducts and metabolites), already described in the literature, were synthesized in order to carry out further investigations. Their ecotoxicity was determined using the standardized Microtox test, and most of the derivatives presented a nontarget toxicity higher than that of diuron. Therefore, the biotransformation of these compounds was tested with four fungal strains and a bacterial strain, which were known to be efficient for diuron transformation. With the exception of the 3,4-dichlorophenylurea, all the degradation products underwent other transformations with most of the strains tested, but no mineralization was observed. For many of them, the biodegradation compound for which the toxicity was important was 3,4-dichlorophenylurea. This study underlines the importance of knowing the nature of the degradation products, which has to be kept in mind while analyzing natural water samples or soil samples. PMID- 11434280 TI - Sediment chemical contamination and toxicity associated with a coastal golf course complex. AB - The increasing density of golf courses represents a potential source of sediment contamination to nearby coastal areas, the chemical and biological magnitude of which is almost unknown. The objective of this study was to determine the concentrations of contaminants and toxicities of sediments impacted by a coastal golf course complex. Sediment contaminant concentrations were determined at least twice during the two-year study period at 14 sampling stations. In addition, a combination of acute and chronic bioassays were conducted exposing four invertebrate test species to whole sediments and associated pore waters. Overall, the Florida, USA, golf course complex had a measurable impact on sediment chemical quality, particularly in near-field areas. Higher concentrations of several trace metals and organochlorine pesticides were detected in many golf course-associated sediments compared with reference areas; however, concentrations decreased seaward and only a few, primarily chlorinated pesticides, exceeded proposed sediment quality guidelines. Chromium, zinc, and mercury were detected more frequently than other trace metals. The DDT and associated metabolites, dieldrin and chlordane, were the more commonly detected organic contaminants. Acute toxicity was uncommon and occurred consistently for sediment collected from one coastal location. In contrast, chronic toxicity occurred at several study sites based on the response of Mysidopsis bahia. It was concluded that the impact of golf course runoff on sediment quality may be subtle and sensitive biological assessment methods, such as chronic toxicity tests, will be needed to detect adverse effects. PMID- 11434281 TI - The power of size. 1. Rate constants and equilibrium ratios for accumulation of organic substances related to octanol-water partition ratio and species weight. AB - Most of the thousands of substances and species that risk assessment has to deal with are not investigated empirically because of financial, practical, and ethical constraints. To facilitate extrapolation, we have developed a model for accumulation kinetics of organic substances as a function of the octanol-water partition ratio (Kow) of the chemical and the weight, lipid content, and trophic level of the species. The ecological parameters were obtained from a previous review on allometric regressions. The chemical parameters, that is, resistances that substances encounter in water and lipid layers of organisms, were calibrated on 1,939 rate constants for absorption from water for assimilation from food and for elimination. Their ratio was validated on 37 laboratory bioconcentration and biomagnification regressions and on 2,700 field bioaccumulation data. The rate constant for absorption increased with the hydrophobicity of the substances with a Kow up to about 1,000 and then leveled off, decreasing with the weight of the species. About 39% of the variation was explained by the model, while deviations of more than a factor of 5 were noted for labile, large, and less hydrophobic molecules as well as for algae, mollusks, and arthropods. The efficiency for assimilation of contaminants from food was determined mainly by the food digestibility and thus by the trophic level of the species. A distinction was made between substances that are stable, that is, with a minimum elimination only, and those that are labile, that is, with an excess elimination probably largely due to biotransformation. The rate constant for minimum elimination decreased with the hydrophobicity of the substance and the weight of the species. About 70% of the variation was explained by the model, while deviations of more than a factor of 5 were noted for algae, terrestrial plants, and benthic animals. Labile substances were eliminated faster than isolipophilic stable compounds, but differences in laboratory elimination and accumulation were small compared with those in field accumulation. Excess elimination by vertebrates was faster than by invertebrates. Differences between terrestrial and aquatic species were attributed to water turnover rates, whereas differences between trophic levels were due to the food digestibility. Food web accumulation, expressed as organism organic solids and organism-food concentrations ratios could be largely explained by ecological variables only. The model is believed to facilitate various types of scientific interpretation as well as environmental risk assessment. PMID- 11434282 TI - The power of size. 2. Rate constants and equilibrium ratios for accumulation of inorganic substances related to species weight. AB - Most of the thousands of substances and species that risk assessment has to deal with are not investigated empirically because of financial, practical, and ethical constraints. To facilitate extrapolation, we have developed a model for concentration kinetics of inorganic substances as a function of the exposure concentration of the chemical and the weight and trophic level of the species. The ecological parameters and the resistances that substances encounter during diffusion in water layers were obtained from previous reviews. The other chemical parameters (the resistances for permeation of lipid layers) were calibrated in the present study on 1,062 rate constants for absorption from water, for assimilation from food, and for elimination. Data on all elements and species were collected, but most applied to aquatic species, in particular mollusks and fish, and to transition metals, in particular group IIB (Zn, Cd, Hg). Their ratio was validated on 92 regressions and nine geometric averages, representing thousands of (near-)equilibrium accumulation ratios from laboratory and field studies. Rate constants for absorption and elimination decreased with species weight at an exponent of about -0.25, known from ecological allometry. On average, uptake-rate constants decreased with about the reciprocal square root of the exposure concentration. About 71 and 30% of the variation in absorption and elimination was explained by the model, respectively. The efficiency for assimilation of elements from food appeared to be determined mainly by the food digestibility and the distribution over egested and digested fractions. (Near )equilibrium accumulation and magnification ratios also decreased with the reciprocal square root of the exposure concentration. The level of the organism solids concentrations ratios roughly varied between one and two orders of magnitude, depending on the number of elements and species groups investigated. Metal concentrations did not increase at higher trophic levels, with the exception of (methyl-)mercury. Organism-solids concentration ratios for terrestrial species tended to be somewhat lower than those for their aquatic equivalents. Food web accumulation, expressed as organism-organic solids and organism-food concentrations ratios, can therefore be only partly explained by ecological variables. The model is believed to facilitate various types of scientific interpretation as well as environmental risk assessment. PMID- 11434283 TI - The use of acute and chronic bioassays to determine the ecological risk and bioremediation efficiency of oil-polluted soils. AB - To compare the effectiveness of acute and chronic bioassays for the ecological risk assessment of polluted soils, soil samples from a site with an historical mineral oil contamination (< 50-3,300 mg oil/kg dry soil) at the Petroleum Harbour in Amsterdam, The Netherlands, were screened for ecological effects using acute and chronic bioassays. A two-step 0.001 M Ca(NO3)2 extraction at a final solution-to-soil ratio of 1:1 was used to prepare extracts for the acute bioassays. Acute bioassays (< or = 5 d) applied to the 0.001 M Ca(NO3)2 extracts from the polluted and reference soils included growth tests with bacteria (Bacillus sp.), algae (Raphidocelis subcapitata), and plants (Lactuca sativa), immobility tests with nematodes (Plectus acuminatus), springtails (Folsomia candida), and cladocerans (Daphnia magna), and the Microtox test (Vibrio fischeri). Chronic bioassays (four weeks) performed on the same soil samples included tests with L. sativa, F. candida, and earthworms (Eisenia fetida) and the bait-lamina test (substrate consumption). The acute bioassays on Microtox showed a response that corresponded with the level of oil pollution. All other acute bioassays did not show such a consistent response, probably because pollutant levels were too low to cause acute effects. All chronic bioassays showed sublethal responses according to the contaminant levels (oil and in some soils also metals). This shows that chronic bioassays on soil samples are more sensitive in assessing the toxicity of mineral oil contamination in soil than acute bioassays on soil extracts. A pilot scale bioremediation study on soils taken from the two most polluted sites and a control site showed a rapid decline of oil concentrations to reach a stable level within eight weeks. Acute bioassays applied to the soils, using Microtox, algae, and D. magna, and chronic bioassays, using plants, Collembola, earthworms, and bait-lamina consumption, in all cases showed a rapid reduction of toxicity, which could be attributed to the degradation of light oil fractions. PMID- 11434284 TI - Ubiquitous observations of enhanced solid affinities for aromatic organochlorines in field situations: are in situ dissolved exposures overestimated by existing partitioning models? AB - This paper investigates the ability of the traditional organic matter partitioning (OMP) model to predict the solid-water distribution, and hence the dissolved exposures, of hydrophobic organic compounds (HOCs) in real field situations. Observed organic-carbon-normalized partitioning coefficients (Koc)obs) of polychlorinated biphenyls, polychlorinated benzenes, polychlorinated dibenzo-dioxins and -furans, and p,p'-dichlorodiphenyltrichlorethane (DDT) with metabolites were selected from the literature and compared with their respective OMP model estimates. For all compound classes and in a majority of the investigated cases, (Koc)obs values were significantly larger than predicted. This translated into factors of overestimated dissolved exposures ranging from 1 to 1,000. Various reasons are discussed for the discrepancies between predictions and actual observations, such as the effect of the diagenetic state and other properties of the particulate organic matter. The greater enhancement in (Koc)obs of planar over nonplanar compounds suggests in certain cases that efficient interactions with aromatic soot phases may be significant. For an improved predictability of (Koc)obs and dissolved exposures of HOCs in the real environment, the inclusion of soot and possibly other distinct subfractions of bulk organic carbon into an extended solid-water partitioning model may be considered. PMID- 11434285 TI - Characterization of enzymes involved in biotransformation of polycyclic aromatic hydrocarbons in terrestrial isopods. AB - Little is known about the capacity of terrestrial invertebrates to transform organic soil pollutants such as polycyclic aromatic hydrocarbons (PAHs). Studies were designed to characterize microsomal mixed function oxygenase and accompanying conjugation enzymes from the hepatopancreas of the terrestrial isopods Porcellio scaber and Oniscus asellus using pyrene and 1-hydroxypyrene as model substrates. The hydroxylation of pyrene and the formation of pyreneglucoside and pyrenesulfate appeared to be sensitive measures for the activity of cytochrome P450 aryl hydrocarbon hydroxylase (AHH), uridinediphosphateglucosyltransferase (UDPGT), and aryl sulfotransferase (ST), respectively. Treatment with the antibiotic riphampicine demonstrated that the enzyme activities originate from the animals themselves and not from symbiotic microflora present in the hepatopancreas and the gut. In both species, ST has a very high affinity for 1-hydroxypyrene with Km values two orders of magnitude lower than that of UDPGT. The Vmax values of UDPGT, however, are 10- to 20-fold higher than that of ST. Taking the P450 activities into consideration, both species are expected to transform PAHs in an equally effective way. When the isopods were fed with food containing benz[a]pyrene and 3-methyl-cholanthrene, none of the enzyme activities appeared to be inducible except for a small enhancement of UDPGT in O. asellus. Our findings indicate that terrestrial isopods have a high, noninducible capacity for biotransformation of PAHs and that the sulfate conjugation pathway is as important as the carbohydrate conjugation pathway. This conclusion is consistent with the low body residues of parent PAHs found in the field. PMID- 11434286 TI - Loss rates of polycyclic aromatic hydrocarbons from sediment and deposit-feeder fecal pellets. AB - Deposit feeders modify sediment by aggregating individual particles into fecal pellets. Loss of contaminants may be either more (enhanced bacterial degradation) or less (hindered diffusion and desorption) rapid for sediment incorporated into pellets. We studied the loss of 10 polycyclic aromatic hydrocarbons (PAHs) from historically contaminated estuarine sediment and the same sediment after it had been pelletized by Capitella sp. I, a polychaete worm typically found in disturbed environments. Fecal pellets initially had higher concentrations of organic carbon and PAHs (enrichment factors 1.03-1.21) due to selective feeding. Over 60 d, desorption of PAHs was minimal, with an average loss of 4 to 5%. Biodegradation accounted for an additional 28% loss. A two-component model fit the data well and indicated that the slowly desorbing fraction was close to one for all PAHs. For low molecular weight PAHs, biodegradation rate constants were greater than desorption rate constants; for high molecular weight PAHs, the opposite was found. Desorption rate constants were similar for fecal pellets (0.0002-0.037/d) and sediment (0.0002-0.031/d). Biodegradation rate constants were also similar for fecal pellets (0.0006-0.022/d) and sediment (0.0002 0.018/d). Thus, incorporation of sediment into robust fecal pellets (half-life on the order of decades) did not affect the loss rates of PAHs. PMID- 11434287 TI - Aircraft and runway deicers at General Mitchell International Airport, Milwaukee, Wisconsin, USA. 1. Biochemical oxygen demand and dissolved oxygen in receiving streams. AB - Aircraft and runway deicers are used during cold weather at many of the world's airports to facilitate safe air travel. Propylene glycol-, ethylene glycol-, and urea-based deicers are known to have very high biochemical oxygen demand. At General Mitchell International Airport (GMIA) in Milwaukee, Wisconsin, USA, deicer application, water chemistry, and dissolved oxygen (DO) data were collected for two deicing seasons in order to evaluate and define premanagement water quality parameters prior to the implementation of a glycol management program. Calculations using stream-monitoring data during a controlled release of deicer provided an estimate of 0.8/d for the first-order decay rate constant, substantially higher than published laboratory test results. For eight precipitation events with deicing activities, between 2.4 and 99% of propylene and ethylene glycol applied to aircraft was delivered directly to receiving streams. The percentage of glycol runoff during an event increased with increasing storm-flow volume. Elevated concentrations of glycol and biochemical oxygen demand were measured downstream from the airport. However, the frequency of low DO concentrations in the receiving streams is comparable with that at an upstream reference site. This is possibly due to slowed bacteria metabolism at low water temperatures, short travel times, and dilution from downstream tributaries. PMID- 11434288 TI - Aircraft and runway deicers at General Mitchell International Airport, Milwaukee, Wisconsin, USA. 2. Toxicity of aircraft and runway deicers. AB - Streams receiving runoff from General Mitchell International Airport (GMIA), Milwaukee, Wisconsin, USA, were studied to assess toxic impacts of aircraft and runway deicers. Elevated levels of constituents related to deicing (propylene glycol, ethylene glycol, and ammonia) were observed in stream samples. The LC50s of type I deicer for Ceriodaphnia dubia, Pimephelas promelas, Hyalela azteca, and Chironimus tentans and the EC50 for Microtox were less than 5,000 mg/L of propylene glycol. Concentrations up to 39,000 mg/L were observed at airport outfall sites in samples collected during deicing events. The IC25s of type I deicer for C. dubia and P. promelas were less than 1,500 mg/L of propylene glycol. Concentrations up to 960 mg/L were observed in low-flow samples at an airport outfall site. Measured toxicity of stream water was greatest during winter storms when deicers were applied. Chronic toxicity was observed at airport outfall samples from low-flow periods in the winter and the summer, with the greater toxic impacts from the winter sample. All forms of toxicity in stream water samples decreased as downstream flows increased. PMID- 11434289 TI - On the validity of classifying chemicals for persistence, bioaccumulation, toxicity, and potential for long-range transport. AB - It is argued that chemical substances can be meaningfully ranked or classified according to their persistence (P), bioaccumulation (B), toxicity (T), and potential for long-range transport (LRT) only if these attributes can be shown to be intensive, as distinct from extensive, properties of the substance, i.e., they are independent of quantity of substance. It is shown that P, B, and LRT can be considered intensive or quasi-intensive properties, but toxicity is more problematic. To obtain an intensive metric of toxicity requires selection of one of several possible extensive quantities that define exposure or dose. Ranking of a group of chemicals by toxicity is shown to be very dependent on which quantity is selected. It is suggested that toxicity metrics, such as lethal concentration to 50% of the population (LC50), lethal dose to 50% of the population (LD50), and threshold limit value (TLV) suffer the severe disadvantage of being dependent on the efficiency of delivery of the substance to the site(s) of toxic action in the organism. The use of measured or calculated internal dose is a preferable measure of toxicity since it reduces ambiguities inherent in the other metrics. Also, the primary concern is not the quasi-intensive property of toxicity; rather, it is the risk of toxic effects, an extensive quantity. To adequately assess the risk of toxic effects, both the toxic hazard and the degree of exposure must be characterized. Since exposure cannot be estimated without knowledge of the emission rate of chemicals to the environment, a compelling case can be made that screening to identify priority P, B, T, and LRT substances should be expanded to include quantity released to the environment as an additional factor. PMID- 11434290 TI - Monitoring river sediments contaminated predominantly with polyaromatic hydrocarbons by chemical and in vitro bioassay techniques. AB - Extracts of sediment samples collected from the Morava River and its tributaries (Czech Republic) were examined for mutagenic, dioxin-like, and estrogenic activities. Moreover, the human leukemic HL-60 cell line was tested as a potential model for the detection of effects of environmental contaminants on cell proliferation and differentiation processes. Analytical data indicate that the sediments were contaminated predominantly with polycyclic aromatic hydrocarbons (PAHs) and phthalate esters. The sums of concentrations of 16 U.S. Environmental Protection Agency priority PAHs ranged from 0.8 to 13.2 micrograms/g and those of phthalates reached up to 3,000 ng/g, while only low levels of chlorinated hydrocarbons were found. The main goal of the present study was to determine effects of PAH prevalence on in vitro bioassays, with special emphasis on dioxin-like activity. The dioxin-like activity was tested using a reporter gene assay based on chemical-activated luciferase expression (the CALUX assay). Significant dioxin-like activity (2.6-40.1 micrograms/g benzo[a]pyrene equivalents and 5.9-48.2 ng/g 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents) was detected in all samples, and the results obtained with various exposure times or with both crude and PAH-deprived extracts indicate that the response was probably caused almost exclusively by the presence of high concentrations of PAHs. This corresponds with results of chemical analyses and indicates that various exposure times would allow a discrimination between dioxin-like activities of persistent compounds and easily metabolized aryl hydrocarbon (Ah) receptor inducers. Only sediment extracts containing the highest concentrations of PAHs were mutagenic, as determined by the umu assay. Estrogenic activity was found in several samples (4.75-22.61 pg/g estradiol equivalents) using cells stably transfected with an estrogen-responsive element linked to a luciferase promoter. Noncytotoxic doses of extracts had no effects on HL-60 cell proliferation, while two of the tested crude extracts significantly enhanced their all-trans retinoic acid-induced differentiation. These activities were not associated with phthalate esters and/or PAHs. Our results indicate that cellular and biochemical in vitro assays based on various specific modes of action may yield data complementary to results of mutagenicity tests and that they could be useful in environmental risk assessment. High levels of PAHs are apparently associated with dioxin-like and mutagenic activities rather than with estrogenic activity. PMID- 11434291 TI - Fluoride concentrations in antler bone of roe deer (Capreolus capreolus) indicate decreasing fluoride pollution in an industrialized area of western Germany. AB - In order to reconstruct temporal changes in ambient fluoride levels in the industrialized Ruhr area (western Germany), we analyzed the bone fluoride content of 167 antlers of roe deer (Capreolus capreolus) killed between 1951 and 1999 in the northern part of this region. Individual values ranged between 110 and 8,178 mg F-/kg ash, and there was an overall marked decrease over the sampling period. Average bone fluoride concentrations in antlers from the periods 1980 through 1989 (geometric mean [95% confidence interval]: 1,490 [1,193-1,861] mg/kg ash) and 1990 through 1999 (753 [644-882] mg/kg ash) differed significantly (p < 0.001) and were both significantly (p < 0.001) lower than those from the periods 1951 through 1969 (3,720 [3,227-4,288] mg/kg ash) and 1970 through 1979 (2,573 [2,203-3,006] mg/kg ash). The findings are seen as indicative of a progressively reduced atmospheric fluoride deposition into the study area, caused by effective emission-control measures in Germany and neighboring countries. Because antlers are replaced annually, grow during a fixed period of some months, and are regularly collected and kept as trophies, they are well suited as monitoring units for analyzing temporal trends in environmental pollution by fluoride and other bone-seeking pollutants. PMID- 11434292 TI - Acute sensitivity of nematode taxa to CuSO4 and relationships with feeding-type and life-history classification. AB - The acute sensitivity to CuSO4 of a broad range of nematode taxa was analyzed in order to assess the potential of changes to nematode community structure to serve as a practical tool for the bioindication of heavy-metal pollution. An easy-to use experimental set-up was developed along with an appropriate mathematical response model in order to quantify the response characteristics of nematodes to CuSO4 pressure. Three similar experiments were conducted using water, dune sand, and sandy soil as media, each of which was subjected to 12 increasing concentrations of CuSO4. In total, 130 response curves representing 70 nematode taxa were produced and analyzed. CuSO4 concentrations were normalized for differences in efficiency among media due to different adsorption. At low CuSO4 concentrations, many taxa exhibited stimulation rather than inhibition regarding recovery efficiency. At higher concentrations, the concentration level at which 50% of the nematode population was recovered after a 24-h incubation (recovery concentration 50% [RC50]) varied widely among taxa and ranged from 0.01 to 4 mM/L CuSO4 (normalized to water). Stimulation of recovery efficiency and RC50 were negatively correlated with the colonizer-persister (C-P) classification of taxa, which discriminates nematodes according to their reproductive potential. The maturity index, which relates to a nematode community's state of disturbance and eutrophication, was negatively correlated with CuSO4 concentration. The properties of the applied test method are discussed as are the relationships between the investigated short-term toxicity effects and long-term toxicity processes in the field. From the large range of observed RC50 values, it is concluded that a meaningful sensitivity classification of nematodes should be possible and thus would allow for a sensitive bioindication of heavy-metal pollution. From the correlation between RC50 and C-P classification, the authors further conclude that the maturity index will, in addition to enrichment and disturbance, respond to heavy-metal pollution and thus may serve as a general indicator of soil health. PMID- 11434293 TI - Reproductive, biochemical, physiological, and population responses in perch (Perca fluviatilis L.) and roach (Rutilus rutilus L.) downstream of two elemental chlorine-free pulp and paper mills. AB - Perch (Perca fluviatilis L.) and roach (Rutilus rutilus L.) populations in a state of late vitellogenesis were studied downstream of two pulp and paper mills and at upstream references in southern Lake Saimaa, Finland. The mills used elemental chlorine-free bleaching and activated sludge effluent treatment technologies. The exposure of fish to pulp mill effluents, as measured by concentrations of chlorophenolics in the bile and liver ethoxyresorufin-O deethylase (EROD) activity, was low and almost similar to the references. Nevertheless, bile resin acid concentrations in exposed perch and roach (260-320 micrograms/ml) and bile beta-sitosterol concentrations in exposed roach (1.5-3.5 micrograms/ml) were, respectively, 10 to 30 times and 2 to 5 times higher compared with the references. Reproductive parameters like plasma 17 beta estradiol and testosterone levels were lower in exposed male and female perch but the same in roach. Vitellogenin concentrations in plasma and liver cytosol of roach were similar. Gonad weight and fecundity were lower in exposed female perch but similar in roach. The relative liver weight was higher in exposed roach but not in perch. The body condition and immunologic parameters in fish were the same. The growth and age at maturity of exposed perch and roach were similar to the references. The size and age distribution of perch in the recipient of one of the mills was shifted toward smaller and younger fish. Our results show that several reproductive variables were altered in late vitellogenic perch in the vicinity of the mills, but they were not altered in roach. PMID- 11434294 TI - Effects of parathion on acetylcholinesterase, butyrylcholinesterase, and carboxylesterase in three-spined stickleback (Gasterosteus aculeatus) following short-term exposure. AB - The sensitivity of butyrylcholinesterase (BChE) toward the inhibition by the organophosphorus insecticide (OP) parathion-ethyl was compared with that of other esterases in the fish three-spined stickleback. Earlier field and in vitro results had suggested the higher sensitivity to OPs of stickleback BChE when compared with acetylcholinesterase (AChE). In the present study, stickleback were exposed in vivo under environmentally realistic conditions using a short duration of exposure (1 h) and parathion concentrations of 0.01, 0.1, and 1.0 microgram/L. Seventy and 80% of nominal concentrations, respectively, were measured in the 0.01 and 0.1 microgram/L treatments. Following exposure, stickleback were maintained in clean water for 48 h (recovery), allowing the metabolic activation of parathion. After recovery, the activities of BChE (axial muscle, gills, liver), AChE (brain, axial muscle, gills), and carboxylesterase (CaE, liver) were determined. Following exposure to 1 microgram/L parathion, the BChE activity was significantly decreased in liver (approximately 60%) and axial muscle (approximately 30%), while its decrease in gills (approximately 30%) was not significant. No effects on BChE activity were observed with 0.1 and 0.01 microgram/L parathion. The AChE and CaE activities remained unaffected with all parathion concentrations used. The results are discussed with respect to the potential application of stickleback BChE as a biomarker of OP exposure. PMID- 11434295 TI - Aquatic reclamation in the Athabasca, Canada, oil sands: naphthenate and salt effects on phytoplankton communities. AB - Microcosm experiments with natural indigenous phytoplankton communities were conducted to assess the effects of waters from oil sands extraction processes, emphasizing the naphthenate and salt constituents. Process waters of varying ages (zero to eight years) remediation histories, and chemical composition were obtained from outdoor mesocosms and inoculated with phytoplankton assemblages from a reference lake in the study area. Community composition measures, including percentage model affinity (PMA) and canonical correspondence analysis (CCA), revealed significant community effects of water from systems less than five years old, with naphthenate concentrations greater than 20 mg/L, compared to water from the reference lake. Canonical correspondence analysis, PMA, and regression analyses further showed that naphthenate concentration was significantly correlated with community structure. Using CCA, groups of taxa characteristic of waters with > 20 mg/L naphthenates (including Botryococcus braunii, Gloeococcus schroeteri, Cosmarium depressum, Chrysococcus rufescens, Chromulina spp., Ochromonas spp., and Keratococcus spp.) were identified. Salinity, as reflected in conductivity, was positively correlated with naphthenate concentration and itself appeared to influence the community structure. The results confirmed an important role for naphthenates in ecological effects of process waters from oil sands mining, but the influence of covarying factors such as salinity requires further investigation. PMID- 11434296 TI - Phototoxicity of azaarene isomers to the marine flagellate Dunaliella tertiolecta. AB - Many studies have shown that narcosis or baseline toxicity of polycyclic aromatic hydrocarbons (PAHs) is strongly related to their lipophilicity. For azaarenes, such relationships have also been demonstrated, but for some compounds, deviations from these relationships have been observed, even for closely related compounds such as isomers. In the present study, the toxicity of four azaarene isomer pairs to the marine flagellate Dunaliella tertiolecta was determined. For quinoline, isoquinoline, acridine, phenanthridine, benz[a]acridine, and benz[c]acridine, the 72-h median effective concentrations for growth were 571, 464, 2.10, 14.7, 0.50, and 0.11 microM, respectively. For the five-ringed isomers dibenz[a,i]acridine and dibenz[c,h]acridine, no effects were observed at the highest concentration tested (0.1 and 0.005 microM, respectively). Growth inhibition by the two-, three-, and four-ringed isomer pairs to D. tertiolecta was well described by molecular volume and log Kow, indicating a narcotic mode of action. However, the toxicity of acridine and benz[c]acridine was much higher than that of their respective isomers, phenanthridine and benz[a]acridine, suggesting an additional specific mode of action. Based on the differences in energies between the highest occupied molecular orbital and the lowest unoccupied molecular orbital, acridine and benz[c]acridine are susceptible to ultraviolet (UV) radiation, in contrast to the other tested compounds. Because UV was present, it is therefore argued that the toxicity of both compounds was photoenhanced. Photoenhanced toxicity may increase the environmental hazard of azaarenes, indicating the importance of enlarging the present monitoring of PAHs with phototoxic N-heterocyclic PAHs and incorporating this mode of action in water-quality criteria. PMID- 11434297 TI - Annual and regional variation in mercury concentrations in wood stork nestlings. AB - Mercury concentrations were measured in blood, down, and feathers from approximately 300 wood stork nestlings in one South Carolina, USA, and four Georgia, USA, colonies from 1996 to 1999. Coastal nestlings generally had lower mercury concentrations than those from inland colonies. Inter-year differences were also apparent, particularly for coastal colonies, where nestling mercury concentrations were higher in 1998 than in 1997 or 1999. In 1998, a wet winter followed by a dry spring and summer produced ideal freshwater foraging conditions and mercury concentrations in coastal nestlings were higher than during the two dry years. There was little inter-year variation in mercury concentrations in nestlings from inland colonies, as parent storks from these colonies forage exclusively in freshwater habitats regardless of rainfall patterns. These results suggest that greater risk of mercury exposure to nestlings is associated with use of freshwater foraging habitats. PMID- 11434298 TI - The effect of thiolate organic compounds on methylmercury accumulation and redistribution in sheepshead minnows, Cyprinodon variegatus. AB - Methylmercury (CH3Hg(II)) was measured in various tissues and whole body of sheepshead minnows; Cyprinodon variegatus, following exposure to 100 ng/L methylmercury chloride (CH3HgCl) alone or in combination with one of the organic compounds cysteine (CH3Hg-Cys), thiourea (CH3Hg-Thu), or thioglycolate (CH3Hg Thg). Overall, the CH3Hg(II) concentration in sheepshead minnows in all the treatments increased with time and was significantly different from the unspiked controls after 24 h. Addition of the organic compounds increased the bioavailability of CH3Hg(II) in the whole body of the fish after 72 h. In particular, the CH3Hg(II) concentration after CH3Hg-Thg and CH3Hg-Thu exposure was, respectively, approximately 1.3- and 1.6-fold higher than with CH3HgCl exposure. A composite of the CH3Hg(II) concentrations in the visceral organs (gill, liver, and intestine) highlighted the effect of the organic compounds, where concentrations in all mercury-thiolate treatments were significantly higher than CH3HgCl alone after 72 h. The most dramatic changes in the different tissues measured were observed in the liver, where the CH3Hg(II) concentrations in the sulfur treatments were significantly higher than the CH3HgCl treatment after 72 h. The results of this study suggest that the CH3Hg-thiolate complexes were bioavailable to sheepshead minnows and effectively induced CH3Hg(II) uptake in tissues at environmentally realistic CH3HgCl concentrations. PMID- 11434299 TI - Incomplete ecdysis is an indicator of ecdysteroid exposure in Daphnia magna. AB - Daphnia magna were exposed for 21 d to the ecdysteroids, 20-hydroxyecdysone (20 E), the accepted molting hormone, and ponasterone A (PoA), an ecdysteroid found in some crustaceans and many plants. Daphnids were monitored for alterations in molting, fecundity, and survival time. The 20-E elicited no significant effects on molting frequency, and its significant effects on reproduction were only at concentrations (260 nM) associated with premature death caused by incomplete ecdysis. We also examined PoA, which has been reported to have 10x higher affinity for the ecdysone receptor than 20-E. Ponasterone A elicited effects similar to those of 20-E at approximately 10x lower concentrations. This suggests that affinity for the receptor is the major parameter determining activity in vitro and in vivo and that differences in metabolism and elimination in vivo were not significant. The effects of PoA on daphnids mimicked those of 20-E except PoA reduced fecundity in the second generation and 20-E had no effect. Last, both exogenous 20-E and PoA show similar effects, including premature death associated with incomplete ecdysis, and the overall difference in toxicity is mostly likely due to receptor affinity. PMID- 11434300 TI - Fate and effects of esfenvalerate in agricultural ponds. AB - The fate of esfenvalerate was investigated by sampling and chemical analysis after spraying of an artificial pond (25 g a.i./ha) and in the laboratory with [14C]esfenvalerate by trapping of 14CO2 and fractionation of the sediment. The effects were investigated on pelagic communities in enclosures in a natural lake and in the laboratory on surface (Cymatia coleoptrata) and sediment (Chironomus riparius) insects. The latter were used in sediment-plus-water and in water-only tests, measuring effects on emergence and mortality. The measurements in the artificial pond indicated exposure concentrations in the surface microlayer, water column, and sediment of 0.4 microgram/L, 0.05 microgram/L, and 9 micrograms/kg dry weight, respectively, two weeks after application. The degradation studies showed a limited mineralization (26.5%) of [chorophenyl 14C]esfenvalerate during 112 d. Part of the substance was transformed to water soluble compounds (18.1%) or compounds attached to fulvic acids (26.2%), humic acids (14.2%), or nonextractable sediment constituents (8.8%). The formulated product Sumi-Alpha 5 FW caused 100% mortality to Cymatia coleoptrata after surface application of 0.13 g a.i/ha. Effects on zooplankton were recorded at 0.005 microgram/L of esfenvalerate. The 96-h median lethal concentration for first-instar larvae of Chironomus riparius was 0.13 microgram/L, whereas the delayed emergence lowest-observed-effect concentration was 0.8 microgram/L. PMID- 11434301 TI - Effect of copper on hatching and development of larvae of the estuarine crab Chasmagnathus granulata (Decapoda, Brachyura). AB - Ovigerous females of the estuarine crab Chasmagnathus granulata were exposed to 0.05, 0.1, 0.5, and 3 mg/L of copper during the egg incubation period. Regarding egg loss, a higher effect was observed at higher copper concentrations: 80% of the females lost their eggs at 0.5 mg/L, whereas no hatching was seen in those exposed to 3 mg/L. A significant decrease was found in the number of hatched larvae in females exposed to 0.5 mg/L, as was a significant decrease in the duration of the incubation period. In addition, several morphological abnormalities were seen and observed. Hydropsy and atrophy of the dorsal spine, pleon, and maxillipeds occurred at the higher copper concentrations, as found in previous studies with other pollutants. Hyperpigmentation of the cephalothorax and pleon was the only abnormality observed at every concentration assayed. Hypopigmented eyes were also evident. This pathology showed that among all defects observed at 0.5 mg/L, hypopigmented eyes had the highest incidence and might be a specific response to copper. PMID- 11434302 TI - Behavioral effects of ivermectin in a freshwater oligochaete, Lumbriculus variegatus. AB - Ivermectin is a potent antiparasitic drug against nematode and arthropod parasites. In this study, we examined the lethal and sublethal effects of ivermectin in a freshwater oligochaete, Lumbriculus variegatus. The median lethal concentration (LC50) at 72 h after ivermectin exposure was 560 nM. Sublethal endpoints focused on several stimulus-evoked locomotor behaviors: escape reflexes controlled by giant interneuron pathways, swimming and reversal, and crawling. Swimming, reversal, and crawling are controlled by nongiant interneuron pathways. Ivermectin inhibited swimming, reversal, crawling frequency, and crawling speed in a time- and concentration-dependent manner with a mean inhibitory concentration (IC50) at 3 h of 1.1, 16, 91, and 51 nM, respectively. Ivermectin at 0.3 nM also significantly decreased the frequency of helical swimming waves. Picrotoxin, a Cl- channel blocker, antagonized the ivermectin-induced decrease in swimming frequency, crawling frequency, and crawling speed. There were no adverse effects on escape reflex 3 h after exposure to 300 nM ivermectin. Electrophysiological recordings showed that ivermectin had no effects on the conduction velocity of giant fiber systems. The results indicated that locomotor behaviors controlled by nongiant locomotor pathways were more sensitive to ivermectin than pathways controlled by giant interneurons and that Cl- channels may be involved in mediating ivermectin's inhibitory effects. PMID- 11434303 TI - Pesticides and amphibian population declines in California, USA. AB - Several species of anuran amphibians have undergone drastic population declines in the western United States over the last 10 to 15 years. In California, the most severe declines are in the Sierra Mountains east of the Central Valley and downwind of the intensely agricultural San Joaquin Valley. In contrast, coastal and more northern populations across from the less agrarian Sacramento Valley are stable or declining less precipitously. In this article, we provide evidence that pesticides are instrumental in declines of these species. Using Hyla regilla as a sentinel species, we found that cholinesterase (ChE) activity in tadpoles was depressed in mountainous areas east of the Central Valley compared with sites along the coast or north of the Valley. Cholinesterase was also lower in areas where ranid population status was poor or moderate compared with areas with good ranid status. Up to 50% of the sampled population in areas with reduced ChE had detectable organophosphorus residues, with concentrations as high as 190 ppb wet weight. In addition, up to 86% of some populations had measurable endosulfan concentrations and 40% had detectable 4,4'-dichlorodiphenyldichloroethylene, 4,4' DDT, and 2,4'-DDT residues. PMID- 11434304 TI - Cadmium binding to a histidine-rich glycoprotein from marine mussel blood plasma: potentiometric titration and equilibrium speciation modeling. AB - Cadmium-binding parameters (conditional stability constants and carrying capacities) of Mytilus edulis blood plasma histidine-rich glycoprotein (HRG) were investigated by potentiometric titrations using a Cd ion-specific electrode. Titration data were applied to a single-component complexation model and expressed as Scatchard plots that were analyzed using the graphical curve peeling method and the algebraic statistical mechanical method. These sets of binding parameters, derived for the purified HRG, were subsequently entered into the geochemical speciation model MINTEQA2 and then used to simulate the experimental titration, thereby determining which set of log K and CL values best represented the titration data. The Cd binding to HRG was best described by a two-class model with log K values of 7.65 +/- 0.10 and 5.41 +/- 0.06 M-1 and carrying capacities of 6.0 +/- 1.2 and 9.5 +/- 0.4 sites/molecule, respectively. At concentration of total Cd measured in the blood plasma of field-collected mussels (< or = 2 x 10( 7) M), plasma Cd speciation would be dominated by the strong affinity sites of HRG (> 93.5% of total Cd binds to HRG), whereas HRG itself would only be 0.05% saturated with Cd, indicating a high-capacity, apparently nonsaturable Cd transport system. PMID- 11434305 TI - Assessment of sediment contamination by spermiotoxicity and embryotoxicity bioassays with sea urchins (Paracentrotus lividus) and oysters (Crassostrea gigas). AB - Gametes (sperm) and fertilized eggs (embryos) of the Mediterranean sea urchin, Paracentrotus lividus, and the Japanese oyster, Crassostrea gigas, were used to investigate the toxicity of two marine sediments, one polluted by polycyclic aromatic hydrocarbons (PAH) and the other by heavy metals. The sediment samples were freeze-dried for storage, and three different treatments were used for analysis: whole sediment, unfiltered elutriate, and filtered elutriate. The two sediments were toxic to sea urchin spermatozoa but not to oyster spermatozoa, and embryotoxicity was almost always the more sensitive endpoint for toxicity assessment. As a rule, whole sediment was more toxic than the elutriates by nearly two orders of magnitude. With respect to embryotoxicity, the whole sediments and the elutriates of the PAH-contaminated sediment were more toxic to oyster embryos, whereas the elutriates of the sediment polluted by heavy metals had stronger effects on sea urchin embryos. The results confirm that bioassays with Japanese oyster embryos provide a more sensitive appraisal of toxicity in the marine environment than bioassays with other developmental stages. As a whole, Mediterranean sea urchins and Japanese oysters were similar in overall sensitivity and are therefore both equally suited as bioassay organisms, but tests with oysters are more reproducible because of the better performance of the controls. PMID- 11434306 TI - Soil-air exchange model of persistent pesticides in the United States cotton belt. AB - Measurements of organochlorine pesticides (lindane, cis-chlordane [CC], trans chlordane [TC], trans-nonachlor [TN]), dieldrin, p,p' dichlorodiphenyldichloroethylene [DDE], and toxaphene) in Alabama, USA, air and soil were used to assess the soil-air equilibrium status and to identify compounds with significant contributions to observed air burdens. Of the compounds tested, p,p'-DDE and toxaphene showed a significant potential for outgasing, followed by dieldrin and trans-nonachlor, which showed moderate outgasing potentials. Lindane, cis-chlordane, and trans-chlordane were near soil air equilibrium. A fugacity-based, multilayered soil-air exchange model was used to predict temporal trends of chemical in air and soil resulting from reemission of soil residues to a presumed clean atmosphere (maximum emission scenario). Results showed that p,p'-DDE and toxaphene accounted for up to 50% of the observed air burden and that approximately 200 to 600 kg of p,p'-DDE and 3,000 to 11,000 kg to toxaphene are released to the atmosphere each year by soils in Alabama (area = 1.23 x 10(11) m2). High annual net fluxes were also predicted for dieldrin and trans-nonachlor (300-1,100 kg and 150-500 kg, respectively), but these only account for up to approximately 20% of their observed air burdens. PMID- 11434307 TI - A plasmid cloning vector with precisely regulatable copy number in Escherichia coli. AB - We have developed a genetic system allowing for precise regulation of plasmid copy number in Escherichia coli cells. A cloning vector based on this system is described in this article. The pTC lambda 3 plasmid is a lambda replicon, but transcription controlling initiation of plasmid DNA replication starts from the PtetA promoter instead of phage lambda PR promoter. Additionally, activity of PtetA promoter is negatively controlled by the TetR repressor whose gene is located on the same plasmid vector and is induced by an analog of tetracycline, autoclaved chlortetracycline (aCT). Using different concentrations of the inducer it is possible to strictly regulate the copy number of pTC lambda 3 and thus the copy number of a cloned gene. The usefulness of the system for the regulatable production of a protein encoded by a gene inserted into pTC lambda 3 plasmid is demonstrated by dependence of beta-galactosidase activity on the lacZ gene dosage. PMID- 11434308 TI - Substitutions of Thr-103-Ile and Trp-138-Gly in amidase from Pseudomonas aeruginosa are responsible for altered kinetic properties and enzyme instability. AB - Pseudomonas aeruginosa Ph1 is a mutant strain derived from strain AI3. The strain AI3 is able to use acetanilide as a carbon source through a mutation (T103I) in the amiE gene that encodes an aliphatic amidase (EC 3.5.1.4). The mutations in the amiE gene have been identified (Thr103Ile and Trp138Gly) by direct sequencing of PCR-amplified mutant gene from strain Ph1 and confirmed by sequencing the cloned PCR-amplified gene. Site-directed mutagenesis was used to alter the wild type amidase gene at position 138 for Gly. The wild-type and mutant amidase genes (W138G, T103I-W138G, and T103I) were cloned into an expression vector and these enzymes were purified by affinity chromatography on epoxy-activated Sepharose 6B acetamide/phenylacetamide followed by gel filtration chromatography. Altered amidases revealed several differences in kinetic properties, namely, in substrate specificity, sensitivity to urea, optimum pH, and enzyme stability, compared with the wild-type enzyme. The W138G enzyme acted on acetamide, acrylamide, phenylacetamide, and p-nitrophenylacetamide, whereas the double mutant (W138G and T103I) amidase acted only on p-nitrophenylacetamide and phenylacetamide. On the other hand, the T103I enzyme acted on p-nitroacetanilide and acetamide. The heat stability of altered enzymes revealed that they were less thermostable than the wild-type enzyme, as the mutant (W138G and W138G-T103I) enzymes exhibited t1/2 values of 7.0 and 1.5 min at 55 degrees C, respectively. The double substitution T103I and W138G on the amidase molecule was responsible for increased instability due to a conformational change in the enzyme molecule as detected by monoclonal antibodies. This conformational change in altered amidase did not alter its M(r) value and monoclonal antibodies reacted differently with the active and inactive T103I-W138G amidase. PMID- 11434309 TI - Isolation of novel cDNA encompassing the ADU balanced translocation break point in the DiGeorge critical region. AB - DiGeorge syndrome (DGS) is a developmental field defect of the third and fourth pharyngeal pouches that are associated with congenital heart defects, hypoparathyroidism, cell-mediated immunodeficiency, velopharyngeal insufficiency, and craniofacial anomalities. Approximately 90% of patients exhibit monosomy in the 22q11 region. In order to isolate the critical gene responsible for DGS, the cDNA libraries were screened with a probe containing the ADU balanced translocation break point, that is a locus reported in one patient (ADU) caused by a balanced translocation between chromosomes 22 and 2. Out of 10(6) clones, three independent overlapping clones were isolated, which were assumed to have originated from a single transcript, DGCR7. This transcript contained a 175-aa long open reading frame (ORF), encoding an acidic (pI = 5.81) and a proline-rich peptide, which are often found in the activation domain of several transcription factors. Also, it was predicted to be a nuclear protein. Northern hybridization detected an approx 1.9 kb transcript in all fetal and adult tissues tested, with strong expression in the fetal liver and kidney. In the case of adult tissues, strong expression was also detected in areas such as the heart, skeletal muscle, liver, and kidney. PMID- 11434310 TI - Bacteriophage lambda-based expression vectors. AB - Bacteriophage lambda has been in use as a cloning vector for over 25 years, and has been used extensively as an expression vector. The efficiency of packaging and infection, and the simplicity of plaque screening are advantages of lambda as a cloning vector. A number of ingenious modifications help overcome the disadvantages associated with its mode of growth and its size. Some lambda vectors have been designed to be readily converted into plasmids or phagemids, and there are a variety of promoters and fusions that can be used to drive expression of foreign genes. Screening lambda libraries with antibodies or ligands is a powerful way of identifying novel genes. PMID- 11434311 TI - Scale-up of suspension and anchorage-dependent animal cells. AB - Alternative culture processes for laboratory scale-up (to 20 L) are described for both suspension and anchorage-dependent cells. Systems range from simple multiple culture units such as the roller bottle, through stirred suspension and microcarrier unit bioreactors, to highly sophisticated perfusion culture capable of maintaining cells at densities of about 10(8)/mL. Critical parameters in scale up are discussed, and the advantages and disadvantages of each culture system are critically evaluated. PMID- 11434312 TI - Automated fluorescent detection of microsatellite instability. AB - Microsatellites are highly polymorphic repetitive DNA segments dispersed throughout the genome and have been widely used for genetic linkage analysis and allele loss. Instability of microsatellites sequences has been linked to deficiencies in DNA mismatch repair, and is observed in a number of different tumor types. Analysis of microsatellite instability is thought to be a useful clinical tool for cancer diagnosis. Fluorescent detection of microsatellite instability using an automated DNA sequencer holds several distinct advantages over traditional radioactive analysis and electrophoresis, allowing simultaneous analysis of a number of different markers for a large number of samples, high resolution, sensitivity, and clear interpretation of data. In this article we present an established protocol, which has been used successfully to detect microsatellite instability in DNA samples from human tumors and circulating tumor DNA in serum/plasma. PMID- 11434313 TI - Procedures for microencapsulation of enzymes, cells and genetically engineered microorganisms. AB - Methods to microencapsulate enzyme, cells, and genetically engineered cells have been described in this article. More specific examples of enzyme encapsulation include the microencapsulation of xanthine oxidase for Lesch-Nyhan disease; phenylalanine ammonia lyase for pheny, ketonuria and microencapsulation of multienzyme systems with cofactor recycling for multistep enzyme conversions. Methods for cell encapsulation include the details for encapsulating hepatocytes for liver failure and for gene therapy. This also includes the details of a novel two-step method for encapsulation of high concentrations of smaller cells. Another new approach is the detailed method of the encapsulation of genetically engineered Escherichia coli DH5 cells for lowering urea, ammonia, and other metabolites in kidney or, liver failure and other diseases. PMID- 11434314 TI - Production of transgenic mice. AB - A "transgenic" mouse is identified by the integration of a foreign DNA into its genome. Such animals serve as experimental systems for the study of gene expression and are often generated as models for human diseases. Direct microinjection of DNA into the male pronucleus of a mouse zygote has been the method most extensively used in the production of transgenic mice. Our method is subdivided into three sections: First, Preinjection, where the animals, the donor eggs, and the injection tools are presented. Second, Injection, where the egg handling and micromanipulation is described. Third, Postinjection, where the surgical transfer into pseudopregnant females completes the procedural sequence. Additional sections are provided to include the materials we use and to offer a series of technical tips that cover various aspects of the transgenic process. PMID- 11434315 TI - Conditional gene knockout using Cre recombinase. AB - Cre recombinase has become an important instrument for achieving precise genetic manipulation in mice. Many of these desired genetic manipulations rely on Cre's ability to direct spatially and temporally specified excision of a predesignated DNA sequence that has been flanked by directly repeated copies of the loxP recombination site. Success in achieving such conditional mutagenesis in mice depends both on the careful design of conditional alleles and on reliable detection of cre gene expression. These procedures include PCR, immunohistochemistry and the use of a recombination-proficient GFP-tagged Cre protein. PMID- 11434316 TI - Asymmetric total synthesis of (-)-alpha-kainic acid using an enantioselective, metal-promoted ene cyclization. AB - [figure: see text] A short and efficient asymmetric total synthesis of the title compound 1, which is an important neurotransmitter, has been achieved. The synthesis features a metal-promoted, enantioselective ene reaction that provides entry into the kainic acid ring system from very simple precursors. Moreover, the zirconium-mediated Strecker reaction, which represents an outgrowth of earlier amide-to-imine methodology developed in our laboratory, demonstrates remarkable chemoselectivity and stereoselectivity. PMID- 11434317 TI - Spongiadioxins A and B, two new polybrominated dibenzo-p-dioxins from an Australian marine sponge Dysidea dendyi. AB - Two new cytotoxic tetrabromodibenzo-p-dioxins, spongiadioxins A (1) and B (2), were isolated from an Australian marine sponge Dysidea dendyi. The structures of these compounds were established by 1D and 2D NMR spectroscopy, X-ray analysis of the methyl ether of spongiadioxin A (3), and synthesis of the methyl ether of spongiadioxin B (4) from diphenyl ether (9) isolated from Dysidea herbacea. PMID- 11434318 TI - Alkaloids from Porcelia macrocarpa. AB - Three new azaanthracene alkaloids, 6,7-dimethoxycleistopholine (3), 5-hydroxy-6 methoxycleistopholine (4), and 5-hydroxy-6,7-dimethoxycleistopholine (5), along with 14 known alkaloids, including the new natural product 6 methoxycleistopholine (2), were isolated from a CH2Cl2 extract of the branches of Porcelia macrocarpa. PMID- 11434319 TI - Trinuclear N,N-bridged copper(II) complexes involving a Cu3OH core: [Cu3(mu 3 OH)L3A(H2O)2]A.(H2O)x (L = 3-acetylamino-1,2,4-triazolate; a = CF3SO3, NO3, ClO4; x = 0, 2) synthesis, X-ray structures, spectroscopy, and magnetic properties. AB - The reaction of Haat [Haat = (3-acetylamino-1,2,4-triazole)] with aquated Cu(CF3SO3)2, Cu(NO3)2, and Cu(ClO4)2, respectively, in water results in the trinuclear complexes [Cu3(OH)(aat)3(CF3SO3)(H2O)2](CF3SO3) (1), [Cu3(OH)(aat)3(NO3)(H2O)2](NO3).(H2O)2 (2), and [Cu3(OH)(aat)3(ClO4)(H2O)2](ClO4) (3). The synthesis, X-ray structure, and magnetic and spectroscopic properties of the three title complexes are described. The cation of the three complexes is trinuclear with a Cu3OH skeleton which has the N-N diazine grouping of a triazole ring as bridge between each pair of copper atoms. The Cu3OH units have an average Cu-O distance of 1.991(6) (1), 2.000(6) (2), and 2.007(6) (3) A, an average Cu Cu' distance of 3.355(2) (1), 3.341(1) (2), and 3.371(3) (3) A, and an average Cu O-Cu' angle of 114.6(3) degrees (1), 112.4(2) degrees (2), and 115.4(3) degrees (3). The existence of the Cu3OH fragment is confirmed by a pseudotetrahedral oxygen environment, by detection of the OH hydrogen atom, and by stoichiometry. In the trinuclear unit the metal ions show, in the first approximation, a pseudo square-planar pyramidal environment forming a CuN2O3 chromophore; three of the basal positions are occupied by N,N,O aat ligand atoms, the fourth one is occupied by the oxygen of the central OH group, and the apical site is occupied by an oxygen atom from a water molecule in the case of two of the copper(II) atoms and by an oxygen atom from the coordinating anion in the case of the third metal ion. The three compounds exhibit strong antiferromagnetic interaction, with similar J constants [J = -197.7 (1), J = -190.9 (2), J = -198.2 (3) cm-1], reaching complete spin coupling at ca. 75 K (1)/55 K (2)/95 K (3). At very low temperature the magnetic moment (magnetic susceptibility) falls below that expected for one unpaired electron. Magnetic parameters are discussed on the basis of the structural results and compared with those reported in the literature for related trimeric Cu(II) compounds with N-O or N-N peripheral bridges. Solid state EPR spectra of the three complexes recorded at liquid N temperature show axial signals. Crystal data: C14H20Cu3F6N12O12S2 (1) (Mw = 917.16) crystallizes in the monoclinic space group, P2(1)/c, Z = 4, with cell dimensions a = 13.080(2) A, b = 17.202(2) A, c = 13.840(2) A, beta = 92.40(1) degrees, and V = 3111.3(7) A3, Dcalcd = 1.958 Mg m-3; the final agreement values were R1 = 0.0582 and wR2 = 0.1462 for 7107 unique reflections. C12H24Cu3N14O14 (2) (Mw = 779.07) crystallizes in the triclinic space group, P1, Z = 2, with cell dimensions a = 9.647(2) A, b = 9.985(2) A, c = 15.314(2) A, alpha = 84.080(10), beta = 87.694(10), gamma = 65.030(10) degrees, and V = 1330.1(4) A3, Dcalcd = 1.945 Mg m-3; the final agreement values were R1 = 0.0397 and wR2 = 0.0950 for 7728 unique reflections. C12H20Cl2Cu3N12O14 (3) (Mw = 817.92) crystallizes in the monoclinic space group, P2(1)/a, Z = 4, with cell dimensions a = 14.238(5) A, b = 16.387(6) A, c = 11.678(4) A, gamma = 90.45(2) degrees, and V = 2724.6(18) A3, Dcalcd = 1.994 Mg m-3; the final agreement values were R1 = 0.0616 and wR2 = 0.1279 for 4038 unique reflections. PMID- 11434320 TI - Structure, physical, and photophysical properties of platinum(II) complexes containing bidentate aromatic and bis(diphenylphosphino)methane as ligands. AB - This study focuses on a series of PtII(L-L')(dppm)n+ complexes, where dppm is bis(diphenylphosphino)methane and L-L' are C-C' (n = 0), C-N (n = 1), and N-N' (n = 2) aromatic ligands. Structural characteristics are as follows: for [Pt(phen)(dppm)](PF6)2, a N-N' derivative, monoclinic, C2/c, a = 33.583(6) A, b = 11.399(2) A, c = 22.158(4) A, Z = 8; for [Pt(phq)(dppm)](PF6), a C-N derivative, triclinic, P1, a = 11.415(3) A, b = 13.450(3) A, c = 14.210(4) A, Z = 2; for [Pt(phpy)(dppm)](PF6), a C-N derivative, triclinic, P1, a = 10.030(3) A, b = 13.010(2) A, c = 15.066(4) A, Z = 2; and for [Pt(bph)(dppm)], a C-C' derivative, P2(1)/c, a = 17.116(7) A, b = 21.422(6) A, c = 26.528(6) A, Z = 12, where phen is 1,10-phenanthroline, phq is 2-phenylquinoline, phpy is 2-phenylpyridine, and bph is 2,2'-biphenyl. Structural features indicate that the Pt-C bond distance is shorter than the Pt-N bond distance in symmetrical complexes and that the Pt-P bond distance trans to N is shorter than the Pt-P bond trans to C. This is consistent with the 31P NMR spectra where the chemical shift of the P trans to C is approximately 10 ppm less than found for P trans to N. The energy maxima of the metal-to-ligand charge-transfer band for the complexes containing various L L' ligands occur in the near-UV region of the spectrum and fall into the energy series bpy > bph > phen > 2-phpy > 2-ptpy > 2-phq > 7,8-bzq, where bpy is 2,2' bipyridine, 2-phpy is 2-phenylpyridine, 2-ptpy is 2-p-tolylpyridine, and 7,8-bzq is 7,8-benzoquinoline. The emission energy maxima, ascribed to variance in metal perturbed triplet ligand centered emission, commence near 500 nm and follow the series phen > bpy > 7,8-bzq > 2-phpy > 2-ptpy > bph > 2-phq. In general, emission is observed at 77 K and in solution at low temperatures, but the temperature dependence of the emission lifetimes indicates thermal activation to another state occurs with an energy of approximately 1800 cm-1 for the complexes, with the exception of [Pt(bph)(dppm)], which has an activation energy of approximately 2300 cm-1. PMID- 11434321 TI - Synthesis, structure, and magnetic properties of Co2([N(CN)2]4bpym).H2O and M([N(CN)2]2bpym).H2O (M = Mn, Fe, Co; bpym = 2,2'-bipyrimidine). AB - Co2([N(CN)2]4bpym).H2O (1) and M([N(CN)2]2bpym).H2O [M = Mn (2a), Fe (2b), Co (2c); bpym = 2,2'-bipyrimidine] have been synthesized and characterized structurally and magnetically. All of the compounds crystallize in the orthorhombic space group Pnma. The unit cell parameters for 1 are a = 16.1684(5) A, b = 12.9860(3) A, c = 10.4207(3) A, and Z = 4. Compound 1 is a 2-D layered structure with water intercalated between sheets. The sheets are composed of ...M [NCNCN]2-M-bpym-M-[NCNCN]2-M... chains, which are linked together by dicyanamides. 2a-c are isomorphic with the unit cell parameters a = 17.5112(4) A, b = 11.9955(4) A, c = 7.4684(2) A for 2a, a = 17.5814(7) A, b = 11.9453(5) A, c = 7.3292(3) A for 2b, a = 17.8642(2) A, b = 11.9216(2) A, c = 7.2860(2) A for 2c, and Z = 4 for all. They crystallize as chains containing metal centers coordinated to two bridging dicyanamides, one terminal dicyanamide, one terminal chelating bpym, and one water molecule. 2a-c are the first examples of compounds containing terminal and mu-bound dicyanamides in the same structure. The broad maximum in the magnetic susceptibility of 1 could not be fit to any known dimer models. However, the high-temperature data were fit to the Curie-Weiss expression with g = 2.86 and theta = -42 K. 2a-c could best be modeled as uniform 1-D chains with g = 2.04, theta = -0.76, and J/kB = -0.15 K for 2a, g = 2.34, theta = -7.6, and J/kB = -0.42 K for 2b, and g = 2.58, theta = -5.4, and J/kB = -1.42 K for 2c. Because of small exchange coupling throughout the extended networks, no long range magnetic ordering was observed. PMID- 11434322 TI - [Evolution of environmental determinants of health in childhood in the city of Sao Paulo, Brazil]. AB - OBJECTIVE: Two consecutive household surveys undertaken in mid-80s and mid-90s in the city of S. Paulo Brazil, made possible to establish time trends of several child health determinants and indicators as well as to analyse the relationships among them. The study intends to report trends in environmental determinants of child health. METHODS: Random samples of the population aged from zero to 59 months were studied: 1,016 children in the period of 1984-85 and 1,280 children in 1995-96. Both surveys investigated several housing characteristics--materials used in the building, size, occupation density, existence of shower, toilet, running water in the kitchen, and the presence of smoker dwellers--as well as access to water supply, sewage, garbage disposal and pavement of public areas. RESULTS: Improvements from mid-80s to mid-90s are observed regarding both housing characteristics and the implementation of basic services public services. The only indicator showing no improvement was the proportion of children living in slums, near 12% in both surveys. However, housing characteristics in slums showed an impressive improvement in the period between the surveys as well as the access of this population to water supply and garbage disposal services. CONCLUSIONS: Improvements in housing characteristics are consistent with increases in the purchasing power reported in the same period. The expansion of public basic services resulted from both public investments and a significant reduction in population growth. The noticeable increase in the coverage of water supply and garbage disposal in the slums indicates a more equitable delivery of the basic services. The same trend was not seen regarding sewage and street paving, which are not widespread in slum areas. The influence that changes in the physical environment may have exerted on several child health indicators is examined in subsequent articles. PMID- 11434323 TI - [Secular trends in intestinal parasitic diseases of childhood in the city of Sao Paulo, Brazil (1984-1996)]. AB - OBJECTIVE: Data from two consecutive household surveys undertaken in mid-80s and mid-90s allow to characterize and analyse secular trends in infant and child intestinal parasitic diseases in the city of S. Paulo, Brazil. METHODS: The two surveys included random population samples aged from zero to 59 months (1,016 in the period of 1984-85 and 1,280 in 1995-96). Stool samples were collected in both surveys and examined by sedimentation techniques using both unstained and Lugol stained preparations. For each survey, the study of the social distribution of the parasitic diseases took into account tertiles of the per capita family income. For the study of the determinants of secular trends, hierarchical causal models, multivariate regression analyses and calculations analogous to the ones used to assess population attributable risks were applied. RESULTS/CONCLUSIONS: In the time span from the first to the second survey, there was a substantial reduction in the prevalence of all parasites (from 30.9% to 10.7%), helminths in general (from 22.3% to 4.8%), giardiasis (from 14.5% to 5.5%) and two or more species of parasites (from 13.1% to 0.5%). A significant decline in prevalence was observed in all social strata and the inverse association between income and intestinal parasites was kept unchanged in the period. Positive changes in distal (family income and maternal schooling) and intermediate determinants (housing, sanitation, and access to health care) of helminthic disease could substantially explain part of its decline in the period. The decline in giardiasis was attributed to improvement in maternal schooling, housing and sanitation. The duplication in the attendance rate to day care nurseries may have restricted the decline rate in the giardiasis prevalence in the study period. PMID- 11434324 TI - [Secular trends in respiratory diseases of childhood in the city of Sao Paulo, Brazil (1984-1996)]. AB - OBJECTIVE: Data from two consecutive household surveys undertaken in mid-80s and mid-90s allow to characterize and analyse secular trends in infant and child respiratory diseases in the city of S. Paulo, Brazil. METHODS: The two surveys included random population samples aged from zero to 59 months (1,016 in the period of 1984-85 and 1,280 in 1995-96). In both surveys the instant prevalence of high respiratory diseases (above the epiglottis) and low respiratory diseases with or without wheezing were calculated. All sampled children were examined in their household at a random day by trained pediatricians using standardized procedures. The examinations included medical history, past respiratory diseases and a complete physical examination. In both surveys the examinations were distributed throughout a period of almost 12 months to assure a uniform coverage of the various areas of the city along the four seasons of the year. For each survey, the study of the social distribution of the diseases took into account tertiles of the per capita family income. For the study of the determinants of secular trends, hierarchical causal models, multivariate regression analyses and calculations analogous to the ones used to assess population attributable risks were applied. RESULTS/CONCLUSIONS: In the time span from the first to the second survey, there was a substantial increase in the prevalence of both low (from 22.2% to 38.8%) and high respiratory diseases (from 6.0% to 10.0% and from 0.8% to 2.8%, without and with wheezing, respectively). In the case of high respiratory disease and low respiratory disease without wheezing, an increase in prevalence was observed among all social strata, which did not interfere with the slightly less favourable situation of the lower income groups. In the case of low respiratory disease with wheezing, the increase was only observed among middle and low-income groups, being particularly high among the lower income group, yielding a significant inverse gradient between income and respiratory disease. Positive changes in distal (family income and maternal schooling) and in intermediate determinants related to housing characteristics would have resulted in a decline, not an increase, in the prevalence of respiratory diseases in the city. The duplication in the attendance rate to day care nurseries seen in the period could have counterbalanced the positive effect due to socioeconomic and housing variables but would not be enough to explain an increase in the disease. PMID- 11434326 TI - Somatopy of perceptual threshold to cutaneous electrical stimulation in man. AB - Neurological testing tools for measuring and monitoring somatosensory function lack resolution and are often dependent on the clinician testing. In this study we have measured perceptual threshold (PT) to electrical stimulation of the skin and compared it with two-point discriminative ability (TPDA) in 12 control subjects. Tests were made on both sides of the body at American Spinal Injury Association (ASIA) key points on seven spinal dermatomes (C3 (neck), C4 (shoulder), C5 (upper arm), C6 (thumb), T8 (abdomen), L3 (knee), L5 (foot)) and in the mandibular (chin) and maxillary (cheek) fields of the trigeminal (V) nerve. Electrical stimulation (0.5 ms pulse width; 3 Hz) was applied via a self adhesive cathode and an anode strapped to the wrist or ankle. The stimulus intensity was adjusted and PT was recorded as the lowest current at which the subject reported sensation. Sites were tested in random order. Indices for both TPDA and PT differed according to the dermatome tested but there was no correlation between TPDA and PT for any dermatome. There was good correlation between results from equivalent dermatomes on left and right sides for both PT and TPDA. Women frequently had lower mean (+/- S.E.) PTs and better TPDA than men; differences were significant (P < 0.05) for PT on the knee (women, 1.31 +/- 0.15 mA; men, 2.05 +/- 0.26 mA) and the foot (women, 2.90 +/- 0.19 mA; men, 4.13 +/- 0.28 mA) and for TPDA on the thumb (women, 3.8 +/- 0.2 mm; men, 7.8 +/- 1.3 mm) and the knee (women, 17.8 +/- 1.6 mm; men, 27.1 +/- 4.0 mm). Four subjects repeated the experiment on another day and the results correlated well with the first test for PT (r2, 0.62) and TPDA (r2, 0.48). PT differs between dermatomes in a predictable way but does not relate to TPDA. PT is easy to measure and may be a useful assessment tool with which to monitor recovery or deterioration in neuropathies, neurotrauma or after surgery. PMID- 11434325 TI - The effect of progesterone on coronary blood flow in anaesthesized pigs. AB - The present study was designed to investigate the effect of progesterone on the coronary circulation and to determine the mechanisms involved. In pigs anaesthetized with sodium pentobarbitone, changes in left circumflex or anterior descending coronary blood flow caused by intravenous infusion of progesterone at constant heart rate and arterial blood pressure were assessed using an electromagnetic flowmeter. In 14 pigs, infusion of 1 mg h(-1) of progesterone caused an increase in coronary blood flow without affecting left ventricular dP/dtmax (rate of change of left ventricular systolic pressure) and filling pressures of the heart. In a further four pigs, this vasodilatory coronary effect was enhanced by graded increases in the dose of the hormone of between 1, 2 and 3 mg h(-1). The mechanisms of the above response were studied in the 14 pigs by repeating the experiment after haemodynamic variables had returned to the control values observed before infusion. In six pigs, blockade of muscarinic cholinoceptors and adrenoceptors with atropine, propranolol and phentolamine did not affect the coronary vasodilatation caused by progesterone. In the remaining eight pigs, this response was abolished by intracoronary injection of N(omega) nitro-L-arginine methyl ester (L-NAME) even when performed after reversing the increase in arterial blood pressure and coronary vascular resistance caused by L NAME with continuous intravenous infusion of papaverine. The present study showed that intravenous infusion of progesterone primarily caused coronary vasodilatation. The mechanism of this response was shown to involve the endothelial release of nitric oxide. PMID- 11434327 TI - A serial study of heart function during pregnancy, lactation and the dry period in dairy goats using echocardiography. AB - Echocardiography and Doppler measurements were performed to investigate changes in heart dimensions and function during pregnancy, lactation and the non pregnant, non-lactating (dry) period in dairy goats. Eight Swedish domestic goats (Capra hircus) were studied with two-dimensional (2DE), M-mode and continuous wave Doppler measurements. Cardiac dimensions did not differ between periods. The mean cardiac outputs were 35 and 28% greater during pregnancy and lactation, respectively, vs. the dry period. Arterial blood pressure (measured by telemetry) did not differ between reproductive periods before echocardiographic measurements, but became elevated during the measurements, with the smallest rise during lactation. Heart rate was elevated during pregnancy and increased further during echocardiographic measurements, but was not elevated during the other periods. The cranial location of the heart influenced the selection of examination window and caused some difficulties in positioning the ultrasound transducer. This may have led to underestimation of the velocity trace integral and therefore to underestimation of cardiac output. However, our values were within the range of these obtained with dye- and thermodilution methods. The goats tolerated the investigations well during lactation and the dry period, but showed increased blood pressure and heart rate during pregnancy. Therefore, the non-invasive and safe echocardiographic-Doppler method appears to be a good alternative for studies of heart dimensions and function during lactation and the dry period in the dairy goat. PMID- 11434328 TI - IGFBPs down under. PMID- 11434329 TI - Endocrinology and metabolism meet in the heart. PMID- 11434331 TI - Twenty five year index, volumes 1-25, March 1979-December 2000. PMID- 11434330 TI - Comparison of statistical power between 2 * 2 allele frequency and allele positivity tables in case-control studies of complex disease genes. AB - In case-control studies of complex disease genes, allele frequencies or allele positivities at candidate loci or markers are compared between cases and controls. Although 2 x 2 contingency tables based on allele frequency and allele positivity are generally used to perform simple statistical tests (e.g. a comparison of two proportions and a chi2 test), little is known about the difference in power between the two tables. In this study, we investigated the number of subjects required to obtain a power of 1-beta with a significance level of alpha for the allele frequency and allele positivity tables. A large difference in the required number of subjects was found between the two tables. Allele positivity tables were suitable for the detection of susceptibility alleles showing a dominant mode of inheritance (MOI). On the other hand, allele frequency tables were suitable for the identification of susceptibility alleles showing a recessive MOI or a multiplicative MOI. In the case of an additive MOI, a suitable table was determined by combining the frequency of the susceptibility allele and the penetrance. These results imply that there are cases in which true association is detected based on one contingency table and is not detected based on another. A simulation analysis revealed that the type I error rate was not much inflated under the null hypothesis of no association, even when a statistical test was performed twice using both allele frequency and allele positivity tables. In contrast, under the alternative hypothesis, the loss of power was marked when a test was performed once using an unsuitable table. In conclusion, statistical tests should be performed using both tables, without adjustment of multiplicity, in case-control studies of complex disease genes when the study objective is exploratory. PMID- 11434332 TI - 46th annual meeting of the Health Physics Society. 10-14 June 2001, Cleveland, Ohio. Abstracts. PMID- 11434333 TI - American Diabetes Association. 61st scientific sessions. Friday, June 22 - Tuesday, June 26, 2001, Philadelphia, Pennsylvania. Abstract book. PMID- 11434334 TI - Abstracts of the International Conference on Apoptosis. 25-28 May 2001, Athens, Greece. PMID- 11434335 TI - Abstracts of the International Conference on Topoisomerases in Cancer. 30 May-2 June 2001, Athens, Greece. PMID- 11434336 TI - Abstracts of the International Conference on Antioxidants in Cancer Prevention and Therapy. 4-7 June 2001, Athens, Greece. PMID- 11434337 TI - Abstracts of the International Conference on New Anticancer Agents. 9-12 June 2001, Athens, Greece. PMID- 11434339 TI - AACC/CSCC annual meeting. July 29-August 2, 2001, Chicago, Illinois. Abstracts. PMID- 11434338 TI - Abstracts of the International Conference on Invasion and Metastasis. 14-17 June 2001, Athens, Greece. PMID- 11434340 TI - 2001 Congress of Epidemiology. A joint meeting of the American College of Epidemiology, American Public Health Association (Epidemiology Section), Canadian Society for Epidemiology and Biostatistics, and Society for Epidemiologic Research. Toronto, Canada, June 13-16, 2001. Abstracts. PMID- 11434341 TI - Clinical significance of high-level gentamicin resistance in enterococci causing bacteraemia. PMID- 11434342 TI - Antioxidant and antimicrobial properties of some Chinese plant extracts. PMID- 11434343 TI - 6th international symposium on myelodysplastic syndromes. 14-17 June 2001, Stockholm, Sweden. Abstracts. PMID- 11434344 TI - Nutritional modulation of acute- and "chronic" -phase responses. PMID- 11434345 TI - Membrane proteins on the move. AB - The Keystone Symposium on Membrane protein structure/function relationships was held on 5-11 March 2001 in Tahoe City, California, USA. PMID- 11434346 TI - Confidentiality of prescription drug information in the era of computers and managed care. McDonald-Merrill-Ketcham lecture. PMID- 11434347 TI - Drug discovery in the wake of genomics. PMID- 11434348 TI - 'ome on the range. PMID- 11434349 TI - Life after sequencing. PMID- 11434350 TI - WASP homology sequences in baculoviruses. PMID- 11434351 TI - The brain may know more than cognitive theory can tell us: a reply to Ted Parks. AB - In reply to Parks' interpretation of Rock's cognitive theory of illusory figures, we maintain our point of view that such a theory has limited heuristic and explanatory power because it fails to predict subjects' responses in psychophysical tasks. As a result, the theoretical framework defended by Parks is not appropriate for suggesting candidate mechanisms of brain-behaviour function that could underly the phenomenal emergence of such figures. PMID- 11434352 TI - Anatomical illustration. 1951. PMID- 11434353 TI - Glomerular lesions in patients with Churg-Strauss syndrome and the anti myeloperoxidase antibody. AB - We report here 4 patients with Churg-Strauss syndrome (CSS) who had classic symptoms including a history of bronchial asthma, severe eosinophilia and necrotizing vasculitis. The antineutrophil antibody (ANCA) against myeloperoxidase (MPO) titers was elevated (44-877 ELISA units), but the ANCA against proteinase-3 (PR3) was negative in all patients. One case was complicated with systemic inflammatory response syndrome (SIRS) and required plasmapheresis and continuous hemodiafiltration. One other patient clinically showed rapidly progressive glomerulonephritis and had hemodialysis 24 times. Two of 4 patients showed good responses with corticosteroid therapy alone, while 2 patients required the addition of cyclophosphamide. Urinary abnormalities such as proteinuria or microscopic hematuria were found in all patients. Three patients had a decreased glomerular filtration rate (GFR) and renal biopsy specimens obtained from these patients showed crescentic glomerulonephritis. One patient had mild to moderate mesangial-proliferative glomerulonephritis with interstitial eosinophilic infiltration. These findings suggest that renal involvements in CSS may not be as uncommon a disorder as previously considered, especially when MPO ANCA is positive. MPO-ANCA may be associated with the onset of glomerular disorders in CSS. PMID- 11434354 TI - Cost-benefit analysis and prediction of 24-hour proteinuria from the spot urine protein-creatinine ratio. AB - AIM: A prospective cross-sectional study was performed on 170 patients with various glomerular diseases to study the accuracy of predicting 24-hour proteinuria from the spot urine protein-creatinine ratio (Up/Uc). A cost-benefit analysis was performed for the New Zealand health economic system to obtain the best cut-off values for proteinuria. SUBJECTS, METHODS AND RESULTS: Two spot urine samples (Up/Uc1 and Up/Uc2) were collected on the same day as the collection of a 24-hour urine. A randomly chosen subsample of 50 patients provided a second set of urine samples. The correlation and precision of agreement between the two methods were examined. The predictive intervals were calculated for derived 24-hour proteinuria. The level of agreement was evaluated by the Bland-Altman method and concordance analysis. The limits of agreement were evaluated against the clinical limits of agreement. A cost-benefit analysis (CBA) was performed to obtain the optimum operating points on receiver operating characteristic (ROC) curves for the best decision threshold. Correlations of r = 0.97 and 0.99 were observed between Up/Uc1, Up/Uc2 and 24-hour proteinuria, respectively. The 95% predictive intervals were wide. A high concordance correlation coefficient was obtained. The most of the differences between the two methods fell within the clinical limits of agreement. The Up/Uc1 of 0.26 and 3.20 represent the best thresholds to detect normal and nephrotic proteinuria, respectively. CONCLUSIONS: Despite wide confidence intervals, a good correlation and precision of agreement were demonstrated between the two methods across the whole range of proteinuria, regardless of the level of renal function. The difference between the two methods was less than the biological variability in the protein excretion and its measurement, enabling the methods to be used interchangeably. The optimum thresholds for abnormal and nephrotic range proteinuria were obtained. PMID- 11434355 TI - Clinical significance of the fractional excretion of anions in metabolic acidosis. AB - The fractional excretion of anions has been proposed as a new index for the differential diagnosis of metabolic acidosis, identifying the properties of the conjugate base by examining the renal handling of the anion. Here, we investigated clinical significance of the fractional excretion of anions in pathophysiologic diagnosis of metabolic acidosis by measuring urine ammonium (NH4+) excretion, the ratio of A plasma anion gap/delta plasma HCO3- concentration (deltaAG/deltaHCO3-), and fractional excretion of anions in three different groups of metabolic acidosis: acid overproduction (8 patients with lactic acidosis, 8 with diabetic ketoacidosis, 3 with hippuric acidosis following glue sniffing), acid underexcretion (10 patients with chronic renal failure) and normal controls (10 normal volunteers who underwent 3-day NH4Cl loading). As expected, urine NH4+ excretion was higher in overproduction acidosis than in acid loaded normal controls (88.1 +/- 12.3 vs. 54.0 +/- 3.7 mmol/day, p < 0.05), and it was lower in chronic renal failure than in acid-loaded normal controls (12.8 +/- 1.7 vs. 54.0 +/- 3.7 mmol/day, p < 0.05). The fractional excretion of anions had no difference between overproduction acidosis and chronic renal failure (41.2 +/- 42.8% vs. 41.0 +/- 8.1%). However, the fractional excretion of anions showed significant differences between the subgroups in acid overproduction (lactic acidosis, 4.7 +/- 0.3%; diabetic ketoacidosis, 45.8 +/- 3.1%; hippuric acidosis, 126.0 +/- 14.4%; p < 0.05). The ratio of plasma deltaAG/deltaHCO3- also exhibited significant differences between the subgroups in acid overproduction (lactic acidosis, 1.5 +/- 0.1; diabetic ketoacidosis, 1.0 +/- 0.1; hippuric acidosis, 0.3 +/- 0.1; p < 0.05). There was an inverse linear correlation between the fractional excretion of anions and the ratio of plasma deltaAG/deltaHCO3- (r2 = 0.89, p < 0.05). In conclusion, determination of the fractional excretion of anions may provide a useful clue to the differential diagnosis of metabolic acidosis caused by acid overproduction. PMID- 11434356 TI - L-arginine supplementation in young renal allograft recipients with chronic transplant dysfunction. AB - AIMS: L-arginine (LA), the precursor of nitric oxide (NO), was suggested to be beneficial in many forms of renal disease: hypertension, ureteral obstructive nephropathy and cyclosporin A (CsA) nephrotoxicity. METHODS: Thus, we investigated the effects of LA supplementation on renal function, proteinuria and blood pressure (BP) in young renal allograft recipients with chronic renal transplant dysfunction treated with CsA. Eleven CsA-treated renal allograft recipients with chronic transplant dysfunction, aged 11-22 years, were randomly assigned to a 6-week treatment period with placebo (P), followed by 2 subsequent 6-week periods with LA supplementation (0.1 g/kg body weight/day) or a 6-week treatment period with LA, followed by 2 subsequent 6-week periods with P. At the end of each treatment period 24-hour BP recordings were made, and GFR (Inutest), RPF (PAH clearance) and the urinary excretion of protein, albumin, nitrate, cGMP and urea were evaluated. RESULTS: In comparison to placebo, LA treatment did not significantly change GFR, RPF, proteinuria and albuminuria, mean systolic or diastolic BP. The urinary excretion of urea and NO3 increased after LA supplementation (uUrea: LA 26.3 +/- 4.6 compared to P 23.5 +/- 4.7 g/day/1.73 m3, p < 0.05, uNO3: LA 514 +/- 152 compared to P 95 +/- 41 mM/day/1.73 m3, p < 0.05), whereas urinary excretion of cGMP remained unchanged. CONCLUSION: LA supplementation did not improve renal function and did not decrease proteinuria in CsA-treated renal allograft recipients with chronic transplant dysfunction possibly because of inhibition of NO-cGMP forming mechanism. PMID- 11434357 TI - Erythrocyte nitric oxide metabolism in patients with chronic renal failure. AB - AIMS: This study aims to investigate arginine-nitric oxide pathway in chronic renal failure (CRF) and to establish erythrocyte nitric oxide synthase (NOS) and arginase (ARG) activities in patients with CRF. MATERIALS AND METHODS: NOS and ARG activities were measured in erythrocytes from 30 patients with CRF and 12 control subjects. RESULTS: Erythrocyte NOS activity was found to be significantly lower and ARG activity higher in patients with CRF compared with controls. No differences were, however, found between patients with and without hemodialysis. A negative correlation (r = -0.7) was established between ARG and NOS activities in erythrocytes from patients. CONCLUSIONS: Results suggest that erythrocyte NO production is diminished in patients with CRF, possibly due to decreased NOS and increased ARG activities. PMID- 11434358 TI - Relationships between homocysteine and related amino acids in chronic hemodialysis patients. AB - AIMS: Homocysteine (Hcy) has emerged as an important risk factor for atherosclerotic disease. Elevated levels in chronic dialysis patients may contribute to high vascular mortality, but little is known about levels of related amino acids in this group. In an observational study in the clinical setting we sought to document these. METHODS: In 114 hemodialysis patients pre dialysis total plasma homocysteine, vitamin B12 and red blood cell (RBC) folate concentrations were measured. In a subgroup of patients (n = 42), other plasma amino acids were measured pre- and post-dialysis. All patients were routinely taking oral folic acid supplements (1.2 mg per week). RESULTS: Elevated homocysteine concentrations were found in all patients (geometric mean 33.1 umol/l, range 13.8 - 69.2 umol/l, laboratory reference range (RR) 3-13 umol/l). RBC folate levels were high (1223 +/- 54.5 nmol/l mean +/- SE, RR 300 - 710 nmol/l) and inversely related to pre-dialysis plasma Hcy (r = -0.44, p < 0.001). Hcy levels were not related to vitamin B12 levels. A history of vascular disease was not associated with higher concentrations of Hcy. Hcy clearance on dialysis was substantial (mean Hcy reduction 33 +/- 14%). While plasma methionine levels were normal, serine levels were significantly lower than the reference range (59.3 +/- 2.39 umol/l (mean +/- SE, RR 70 - 195 umol/l)) and directly related to levels of glycine (r = 0.52, p < 0.001). Glycine levels were within normal range. Although overall levels were low, higher serine levels were related to elevated homocysteine (r = 0.42, p < 0.01). Dialytic loss of glycine, serine and methionine was moderate. CONCLUSION: An inverse association between RBC folate and homocysteine levels extended to 3 times the upper limit of normal for folate, suggesting a role for high dose folic acid supplementation in the treatment of renal-failure related hyperhomocysteinemia. Low serine levels are expected as it is primarily synthesized in the kidney. The direct relationship between serine and homocysteine is consistent with the reported lack of effect of serine supplements on high Hcy levels. PMID- 11434359 TI - Improvement of thyroid hormone profile and thyrotrophin (TSH) surge alterations in hemodialysis patients on erythropoietin treatment. AB - BACKGROUND, MATERIAL AND METHODS: This study was performed in 20 patients with end-stage chronic renal failure (CRF) and 10 healthy volunteers. All of the patients were on regular hemodialysis treatment (RHD), 10 of whom were on recombinant human erythropoietin (rHuEPO) therapy. Hematocrit levels of the patients with CRF on rHuEPO were between 0.30 to 0.33 and not on rHuEPO were below 0.24. Baseline serum T3, T4, fT3, fT4 and TSH levels were measured and TRH stimulation test was performed in patients and control subjects. Serum TSH levels were measured hourly during the afternoon (2 to 5 p.m.) and at night (10 p.m. to 2 a.m.) to determine the nocturnal rhythm of TSH. RESULTS: The mean T3 in rHuEPO, not rHuEPO and control groups were 98.01 +/- 5.54, 70.55 +/- 7.09, 98.29 +/- 4.2 ng/dl; T4 6.47 +/- 0.68, 6.39 +/- 0.59, 8.35 +/- 0.46 ng/dl; fT3 2.24 +/- 0.19, 1.52 +/- 0.24, 2.29 +/- 0.17 pg/ml and fT4 0.88 +/- 0. 14, 0.75 +/- 0.14, 0.97 +/ 0.10 ng/dl, respectively. These values were significantly lower in patients not on rHuEPO compared to controls (p < 0.05). In patients on rHuEPO only T4 values were lower than in the controls (p < 0.05). In patients not on rHuEPO the T3, and fT3 were significantly lower than the values of patients on rHuEPO treatment (p < 0.05). Normal in 8 (80%), blunted in 1 (10%), no TSH response in 1 (10%) to TRH stimulation were obtained in rHuEPO group. TSH response was normal in 1 (10%), and delayed in 9 (90%) patients not on rHuEPO. The circadian nocturnal rhythm of TSH was abnormal in 8 (80%) patients not on rHuEPO, in 2 (20%) patients on rHuEPO. As a result, CRF and RHD distorts the circadian TSH rhythm and substantially change the thyroid hormone profile probably by affecting hypothalamic-pituitary-thyroid axis. Distortion of the circadian rhythm of TSH and TSH response to TRH points to a defect at the level of hypothalamus and pituitary gland. CONCLUSION: rHuEPO treatment has some beneficial effects on hypothalamo-pituitary-thyroid axis in the patients on RHD. PMID- 11434360 TI - Patient to patient transmission of hepatitis C virus in hemodialysis units. AB - AIMS: In dialysis patients, blood transfusions and long-term dialysis are well known risk factors for transmission of hepatitis C virus. In this study the impact of use of dedicated hemodialysis (HD) units on the anti-HCV conversion rates was studied in patients of two different hemodialysis units in a city, Kayseri, between October 1995, and March 1999. MATERIALS AND METHODS: In the HD Unit of Erciyes University (HUEU), anti-HCV-positive and -negative patients were dialyzed on the dedicated machines in the same big room and seropositive patients for HBsAg in isolated rooms. In the HD Unit of Kayseri State Government Hospital (HUSH) only seronegative for anti-HCV and hepatitis B patients were treated. If a patient became positive, the patient was transferred to HUEU. Seventy-five patients have been receiving hemodialysis therapy in HUEU. Thirteen HBsAg positive and 62 HBsAg-negative patients were dialyzed in separate rooms. Of 62 HBsAg-negative patients, 22 (35.5%) were already positive for HCV antibody when they started dialysis or before the study period. Forty seronegative patients (64.5%) for anti-HCV (23 males, 17 females) were treated with 22 anti-HCV positive patients in the same room in HUEU. The mean duration of dialysis treatment was 24.7 +/- 21.0 months (range 4 to 96 months). Of the 40 patients, 28 (70%) became positive during the study period. Of 28 patients who became seropositive, 10 (35.7%) had a history of blood transfusion. Fifty-four patients (21 males, 33 females) were treated in HUSH during the study period. The mean duration of dialysis treatment was 19.3 +/- 9.6 months (range 5 41). Eight patients (14.8%) became anti-HCV-positive. Of these, 7 had received blood transfusion (88%). RESULTS: The seroconversion rate of patients in HUEU was higher than that of HUSH (odds ratio 3) (p < 0.05). Data derived from our patients showed that contamination appeared to be both transfusional and nosocomial and that there is a possibility of transmitting HCV infection in hemodialyzed patients never submitted to blood or blood products transfusion. Nosocomial spread of HCV in HD units which both seropositive and seronegative patients treated together was higher than that of dedicated unit. This is true even though we separated anti-HCV-positive dialysis machine. PMID- 11434361 TI - Association of a POEMS syndrome and light chain deposit disease: first case report. AB - Monoclonal immunoglobulin (Ig) deposition diseases are characterized by deposition in tissues of excessive amounts of the Ig, compromising organ functions. Light chain deposition disease (LCDD) and AL amyloidosis are the commonest [Buxbaum 1992]. LCDD is usually characterized by rapidly progressive renal failure with glomerular and tubular deposits of Ig fragments mostly composed by kappa light chain. Monoclonal Ig production can also be observed associated with various symptoms, that, taken together, have been described as the Crow-Fukase syndrome or POEMS syndrome. It associates polyneuropathy, organomegaly, endocrinopathy, monoclonal Ig, and skin changes. In POEMS syndrome, renal abnormalities are rare and are reported as a moderate renal insufficiency with mild proteinuria or acute functional renal insufficiency leading in some cases to end-stage renal failure [Fukatsu et al. 1991]. Although a monoclonal Ig is produced, no Ig deposit disease had been described in POEMS syndrome except a case of AL amyloidosis [Toyokuni et al. 1992]. Here, to our knowledge, we report the first case of an LCDD associated with a POEMS syndrome. Although an autologous bone marrow graft was realized, the monoclonal component reappeared and was responsible for end-stage renal disease, cachexia and death. PMID- 11434362 TI - Polycystic horseshoe kidney. AB - Horseshoe kidney is a renal fusion anomaly during embryogenesis. The reported incidence of horseshoe kidney varies from 1 in 400 to 1 in 1,800 live births (commonly quoted 1 per 400) [Wilson and Azmy 1986]. Adult polycystic kidney disease is a hereditary disorder with autosomal-dominant transmission. Its incidence is approximately one in every 1,000-5,000 cases [Levine et al. 1997]. Polycystic horseshoe kidney is a very rare occurrence with incidence ranges of 1 in 134,000 to 1 in 8,000,000 cases [Brum et al. 1997]. We add another case to the literature. PMID- 11434363 TI - IL-6 may be the key mediator in trimethoprim-induced systemic adverse reaction and aseptic meningitis: a reply to Muller et al. PMID- 11434364 TI - Deterioration in renal function associated with fibrate therapy. PMID- 11434365 TI - Coverage policy for dialysis laboratory tests needs review. RPA. Renal Physicians Association. PMID- 11434366 TI - Relation of sleep-disordered breathing to cardiovascular disease risk factors: the Sleep Heart Health Study. AB - Associations between sleep-disordered breathing and cardiovascular disease (CVD) may be mediated by higher cardiovascular risk factor levels in those with sleep disordered breathing. The authors examined these relations in the Sleep Heart Health Study, a multiethnic cohort of 6,440 men and women over age 40 years conducted from October 1995 to February 1998 and characterized by home polysomnography. In 4,991 participants who were free of self-reported CVD at the time of the sleep study, moderate levels of sleep-disordered breathing were common, with a median Respiratory Disturbance Index (RDI) of 4.0 (interquartile range, 1.25-10.7). The level of RDI was associated cross-sectionally with age, body mass index, waist-to-hip ratio, hypertension, diabetes, and lipid levels. These relations were more pronounced in those under age 65 years than in those over age 65. Women under age 65 years with RDI in the higher quartiles were more obese than men with similar RDI. Although the pattern of associations was consistent with greater obesity in those with higher RDI, higher body mass index did not explain all of these associations. If sleep-disordered breathing is shown in future follow-up to increase the risk for incident CVD events, part of the risk is likely to be due to the higher cardiovascular risk factors in those with higher RDI. PMID- 11434367 TI - Clearance of antiphospholipid antibodies in pregnancies treated with heparin. PMID- 11434368 TI - Royal College of Paediatrics and Child Health. Proceedings of the 3rd spring meeting, University of York, 12-15 April 1999. Scientific programme and abstracts. PMID- 11434369 TI - Sonochemistry of carbohydrate compounds. AB - A literature survey of carbohydrate sonochemistry is presented. The basic physical principles are discussed qualitatively before the main applications are reviewed. Along with other non-classical activation methods developed in accordance with 'Green Chemistry' requirements, ultrasonic irradiation offers important potential for the conversion of biomass raw materials such as polymeric carbohydrates to useful lower weight molecules. A number of reactions involving mono- or disaccharides, considered as a renewable source of fine chemicals, can also be performed with ultrasonic activation. This is the case for glycosylation, acetalization, oxidation, C-D, C-heteroatom, and C-C bond formations, which are improved in terms of reaction rates and yields and, in some instances, in their chemo-, regio- and stereoselectivities. The equipment available is briefly described, and practical considerations are given. PMID- 11434370 TI - Synthesis of C-3 nitrogen-containing derivatives of N-acetyl-alpha,beta-D mannosamine as substrates for N-acetylneuraminic acid aldolase. AB - The synthesis of 3-azido-3-deoxy, 3-amino-3-deoxy and 3-N-tert-butyloxycarbonyl-3 deoxy derivatives of 2-acetamido-2-deoxy-alpha,beta-D-mannose (N-acetyl alpha,beta-D-mannosamine, ManNAc), is presented. The 3-azido-3-deoxy- and 3-N tert-butyloxycarbonyl compounds were further characterised as their peracetates. A preliminary study has found that these C-3 nitrogen-substituted derivatives of ManNAc not to be substrates for Neu5Ac aldolase. PMID- 11434371 TI - Di-tert-butyl diethylphosphoramidite as the phosphitylating reagent in the preparation of 3-deoxy-3-C-methylene-D-ribo-hexose-6-phosphate and 3-deoxy-3-C methylene-D-erythro-pentose-5-phosphate. AB - 3-Deoxy-3-C-methylene-D-ribo-hexose-6-phosphate and 3-deoxy-3-C-methylene-D erythro-pentose-5-phosphate were prepared from a common intermediate 3-deoxy-3-C methylene-1,2-O-isopropylidene-alpha-D-ribo-hexofuranose. The preparation of the phosphorylated unsaturated sugars employed di-tert-butyl diethylphosphoramidite as the phosphitylating reagent. The removal of all the protecting groups was done under acidic conditions in the ultimate step. The unsaturated sugar phosphates were competitive inhibitors but neither substrates nor inactivators of glucose-6 phosphate and ribose-5-phosphate isomerases. PMID- 11434372 TI - 2-Nitro and 4-nitro-quinone-methides are not irreversible inhibitors of bovine beta-glucuronidase. AB - 4-Benzylamino-(and 4-chloromethyl)-2-nitro-beta-D-glucuronides (4, 10) and their 2-substituted-4-nitro regioisomers (7, 13) were prepared by glycosidation of the 3-nitro-4-hydroxy- and the 2-hydroxy-5-nitro-benzylic alcohol, respectively, with a glucuronyl donor. Carbonate activation followed by reaction with benzylamine or methanesulfonyl chloride afforded, after complete deprotection, the target molecules 4, 7, 10 and 13. These compounds have been synthesized to determine whether these molecules are (or not) glucuronidase inhibitors. After incubation with bovine liver beta-glucuronidase, none of the cleavage products (the titled quinone-methides) showed to be irreversible inhibitors of this enzyme. PMID- 11434373 TI - Regioselective C-3-O-acylation and O-methylation of 4,6-O-benzylidene-beta-D gluco- and galactopyranosides displaying a range of anomeric substituents. AB - The regioselective C-3-O-acylation and O-methylation of a range of 4,6-O benzylidene-beta-D-gluco- and galactopyranosides has been studied. Regioselectivity is achieved by forming the copper chelate of the 2,3-diol using either sodium hydride and copper(II) chloride, or copper(II) acetylacetanoate, or copper(II) acetate, prior to introduction of the acylating or methylating agent. PMID- 11434374 TI - Structure of a polysaccharide from a Rhizobium species containing 2-deoxy-beta-D arabino-hexuronic acid. AB - The structure of the extracellular polysaccharide (EPS) produced by the Rhizobium sp. B strain isolated from atypical nodules on alfalfa has been determined using a combination of chemical and physical techniques (methylation analysis, high pH anion exchange chromatography (HPAEC), mass spectrometry and 1-D and 2-D NMR spectroscopy). As opposed to the EPS from other strains of Rhizobium, the EPS from the sp. B strain contains D-Glc together with L-Rha and 2-deoxy-D-arabino hexuronic acid. It is a polymer of a repeating unit having the following structure: --> 4)-beta-D-Glcp-(1 --> 4)-alpha-L-Rhap -(1 --> 3)-beta-D-Glcp-(1 - > 4)-2-deoxy-beta-D-GlcpA-(1 -->. The polysaccharide also contains 0.6 O-acetyl groups per sugar which have not been located. PMID- 11434375 TI - Density-labelling of cell wall polysaccharides in cultured rose cells: comparison of incorporation of 2H and 13C from exogenous glucose. AB - Labelling with stable isotopes has under-exploited potential for studies of polysaccharide endotransglycosylation in vivo. Ideally, the labelled polysaccharides should have the highest possible buoyant density. Although [13C6]glucose has previously been used as a precursor, it was unclear whether 2H would be efficiently incorporated from [2H]glucose or lost as D2O. Rose (Rosa sp.) cell-suspension cultures efficiently incorporated 13C from D [13C6,2H7]glucose into wall polysaccharides with negligible dilution from atmospheric 12CO2. Also, approximately 70% of the 2H atoms in D-[13C6,2H7]glucose were retained during polysaccharide biosynthesis. This shows that relatively few cycles of intermediary metabolism leading to the release of D2O occurred before sugar residues were incorporated into wall polysaccharides. In agreement with these observations, isopycnic centrifugation in caesium trifluoroacetate gradients showed that the hydrated buoyant density of xyloglucan synthesised by rose cells growing on [13C6,2H7]glucose and [13C6]glucose was 3.7 and 2.6% higher, respectively, than in isotopically non-labelled cultures. Thus, [13C,2H]glucose-feeding enabled a 42% better resolution of 'heavy' from 'light' xyloglucan than [13C]glucose-feeding. PMID- 11434376 TI - Large-scale preparation of the oligosaccharide phosphate fraction of Pichia holstii NRRL Y-2448 phosphomannan for use in the manufacture of PI-88. AB - Mild acid-catalysed hydrolysis of the extracellular phosphomannan of the yeast Pichia holstii NRRL Y-2448 produces a high-molecular-weight phosphomannan core, a low-molecular-weight oligosaccharide phosphate fraction, and a neutral oligosaccharide fraction. A method was developed for the large-scale preparation of the oligosaccharide phosphate fraction, consisting predominantly of the pentasaccharide phosphate, 6-O-PO3H2-alpha-D-Man-(1 --> 3)-alpha-D-Man-(1 --> 3) alpha-D-Man-(1 C 3)-alpha-D-Man-(1 --> 2)-D-Man, for use in the manufacture of the promising new anti-cancer agent, PI-88. Further insights were also gained into the structure of the phosphomannan by HPLC analysis of the time course of the hydrolysis reaction. PMID- 11434377 TI - Determination of carbohydrates as their p-sulfophenylhydrazones by capillary zone electrophoresis. AB - p-Hydrazinobenzenesulfonic acid was explored as an ultraviolet labeling reagent for capillary electrophoresis of mono-, di- and trisaccharides. The labeling reaction that produces p-sulfophenylhydrazines took less than 8 min, and introduced both chromphore and charged groups into the carbohydrate molecules. The derivatives of nine mono- and disaccharides were completely separated in 9 min using a 100 mM borate buffer at pH 10.24. On-column UV detection at 200 nm allowed the detection of glucose with a mass detection limit of 17.6 fmol or a concentration limit of 3.6 microM. Reproducible quantification of carbohydrates was achieved in the concentration range of 0.1-9.1 mM in reaction solution. The method was applied successfully to determine the monosaccharide composition of laminaran. PMID- 11434378 TI - Metal ion coordination of macromolecular bioligands: formation of zinc(II) complex of hyaluronic acid. AB - The coordination of zinc(II) ion to hyaluronate (Hya), a natural copolymer, in aqueous solution at pH 6 was investigated by potentiometric and circular dichroism (CD) spectroscopic methods, and by monitoring the changes in macroscopic properties by high-precision measurements. The zinc(II)-selective electrode, and CD measurements proved the binding of zinc(II) by Hya. A number of Hya fragments (Mr approximately 3.3 x 10(3)-1.4 x 10(6)) were studied to estimate the contributions of the polyelectrolyte effect, the solvation and host-guest interactions to the extra stabilization of the macromolecular zinc(II) complexes as compared with the monomeric unit. The zinc(II) ion activity increase reflected a stability decrease for the fragments with Mr < 4 x 10(4). This molecular weight differs from that where cleavage of the Hya skeleton starts (approximately 5 x 10(5), according to the size-exclusion gel, and anion-exchange chromatographic behavior of the Hya fragments) and from that where the polyelectrolyte effect stops (approximately 6 x 10(3)). The excess volumes and Bingham shear yield values of the solutions revealed the transformation of the coherent random coil structure stabilized by intermolecular association in the NaHya to an intramolecular association producing the globular structure of the ZnHya molecule, with a smaller but more strongly bound solvate water sheet. PMID- 11434379 TI - Analysis of mono- and oligosaccharides by multiwavelength surface plasmon resonance (SPR) spectroscopy. AB - Surface plasmon resonance (SPR) spectra of different saccharides were collected using a home-made multiwavelength SPR apparatus. Pentoses, hexoses, disaccharides and a trisaccharide were distinguished from one another according to their SPR spectra collected at the same concentration. The spectra were also used for the quantitation of sugars by exploring the linear relationship between resonance wavelength and solute concentration. The dynamic linear ranges for the determination of glucose, sucrose and raffinose are 0.01-0.2, 0.005-0.1 and 0.0025-0.1 mol/L, respectively. The SPR spectrum of a mixture of two components was investigated. While the experiments have not been carried out, the implications from this work are that the technique would be applicable to mixtures containing more than two components. PMID- 11434380 TI - Synthetic studies on dendritic glycoclusters: a convergent palladium-catalyzed strategy. AB - A facile Pd-catalyzed strategy by which multiantennary glycoclusters and sugar dendrons can be readily assembled in one-step is described. PMID- 11434381 TI - LiBF4-mediated C-glycosylation of glycals with allyltrimethylsilane: a facile synthesis of allyl C-glycosylic compounds. AB - The treatment of glycals with allyltrimethylsilane in the presence of lithium tetrafluoroborate in acetonitrile gave the corresponding allyl 2,3-unsaturated C glycosylic compounds in excellent yields with high anomeric selectivity. PMID- 11434382 TI - Homologation of methyl 2-azido- and 2-acetamido-3,4-di-O-benzyl-2-deoxy-D hexopyranosides with allyloxymethylmagnesium chloride. AB - Methyl 2-azido-2-deoxy-hexodialdo-1,5-pyranosides of the alpha-, beta-D-gluco and alpha-D-manno configuration as well as methyl 2-acetamido-2-deoxy-hexodialdo-1,5 pyranosides of the alpha- and beta-D-gluco configuration, protected at positions 3 and 4 with O-benzyl groups were reacted with an excess of allyloxymethylmagnesium or (phenyldimethylsilyl)methylmagnesium chlorides to afford mixtures of C-6 stereoisomeric heptopyranosides. Configuration of the products separated by column chromatography was assigned by 1H NMR data. PMID- 11434383 TI - Crystal structures of cyclomaltohexaose (alpha-cyclodextrin) complexes with p bromophenol and m-bromophenol. AB - Crystal structures of cyclomaltohexose (alpha-cyclodextrin) complexes with p bromophenol and m-bromophenol have been determined by single-crystal X-ray diffraction. The space group of the alpha-cyclodextrin-p-bromophenol complex is P2(1)2(1)2(1) with unit cell dimensions of a = 15.318(3), b = 24.733(3), c = 13.457(2) A, and that of the alpha-cyclodextrin-m-bromophenol complex is P2(1)2(1)2 with unit cell dimensions of a = 25.858(7), b = 27.263(8), c = 8.145(3) A. In crystals, the alpha-cyclodextrin-p-bromophenol complex and the alpha-cyclodextrin-m-bromophenol complex form a layer-type and a channel-type molecular packing structure, respectively. The intermolecular hydrogen-bond interactions of the hydroxyl groups of bromophenols are closely related to the molecular packing structure. PMID- 11434384 TI - Granulocyte-Colony stimulating factor and macrophage-colony stimulating factor in patients with non-small-cell lung cancer. AB - We have investigated the serum level of granulocyte-colony stimulating factor (G CSF) and macrophagecolony stimulating factor (M-CSF) in non-small-cell lung cancer (NSCLC), in relation to the control group and commonly accepted tumor markers, such as carcinoembryonic antigen (CEA) and cytokeratin fragment 19 (CYFRA 21-1). Additionally, we have defined the diagnostic sensitivity, specificity, positive predictive value, negative predictive value and receiver operating characteristics (ROC) curve of G-CSF and M-CSF. Serum levels of cytokines were measured in 61 patients with NSCLC and in 20 healthy subjects. G CSF and M-CSF were determined using ELISA. CYFRA 21-1 was measured by radioimmunoassay and CEA by microparticle enzyme immunoassay. There were significant increases in the level of circulating G-CSF in the lung cancer patients compared to the control group. Moreover, the diagnostic sensitivity of G CSF was higher (56%) than the sensitivity of CYFRA 21-1 (51%), but lower than the CEA sensitivity (62%). The diagnostic specificity of G-CSF was higher (70%) than the M-CSF specificity (40%) and the G-CSF predictive values were higher in relation to the predictive values of M-CSF. These results suggest a potential role of G-CSF as a tumor marker for NSCLC. PMID- 11434385 TI - Circulating matrix metalloproteinases and their inhibitors in premature coronary atherosclerosis. AB - To investigate the clinical significance of circulating matrix metalloproteinases (MMPs) and their tissue inhibitos (TIMPs) in patients with premature coronary atheroscrelosis, we studied 53 consecutive male patients with angiographically defined premature (<65 years) and stable coronary artery disease. Plasma levels of MMP-2, MMP-3, MMP-9, TIMP-1, and TIMP-2 were determined in peripheral blood by a sandwich enzyme immunoassay, and the results were compared with those from 133 age-matched control males. There were significant differences in all the MMPs and TIMPs (p<0.001) between patients and controls. In the patient group, the levels of MMP-9 (mean +/- SD (ng/ml) 27.2 +/- 15.2/21.8 +/- 15.2) and TIMP-1 (130.4 +/- 55.7/94.5 +/- 26.3) were significantly higher, and the levels of MMP-2 (632.5 +/- 191.6/727.6 +/- 171.4), MMP-3 (53.1 +/- 31.2/79.6 +/- 29.9), and TIMP-2 (24.7 +/- 15.2/35.4 +/- 16.4) were significantly lower than those of controls. We found significant positive correlation between plasma MMP-9 levels and low-density lipoprotein (LDL)-cholesterol levels (Rs = 0.168, p = 0.022), and significant negative correlation between plasma MMP-9 levels and high-density lipoprotein (HDL)-cholesterol levels (Rs = -0.164, p = 0.026) by Spearman rank correlation test. In contrast, plasma MMP-2 (Rs = 0.181, p = 0.014) and MMP-3 (Rs = 0.260, p = 0.0004) levels were positively correlated with HDL-cholesterol levels. TIMP-2 levels were negatively correlated with total cholesterol (Rs = -0.197, p = 0.007) and LDL-cholesterol (Rs = -0.168, p=0.022) levels. These results suggest that the circulating levels of MMPs and TIMPs are altered in patients with premature coronary atherosclerosis and that plasma lipoprotein cholesterol levels correlate with these, possibly as a result of the lipoprotein-vessel wall interactions. PMID- 11434386 TI - Real-Time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) for the measurement of prostate-specific antigen mRNA in the peripheral blood of patients with prostate carcinoma using the taqman detection system. AB - Circulating prostate cells can be detected in peripheral blood of patients with clinically localized or advanced prostate carcinoma. Traditionally, nested reverse transcriptase-polymerase chain reaction (RT-PCR) is used for this as a sensitive, but qualitative only, detection system. We developed a quantitative real-time RT-PCR method for measuring prostate-specific antigen (PSA) mRNA in peripheral blood of prostate cancer patients. A quantitative assay was developed using an external standard reference curve generated with RNA from the human prostate cell line LNCaP. Basal blood samples were collected from 44 patients without evidence of distant metastases and from 30 healthy controls. In 29 patients surgically treated with radical prostatectomy, the measurement of PSA mRNA was performed in blood samples collected before, at the end and 6 days after surgery. In 14 patients treated with radiotherapy, the measurements were repeated at 3-month intervals to evaluate time-related changes during therapy. The measurements were also performed for one year at 3-month intervals in one patient treated with anti-androgen therapy. We found detectable PSA mRNA in 14/44 (32%) basal blood samples. A wide range of values were observed in these patients, ranging from 0.5 to 1724 pg of total LNCaP RNA/ml blood. In patients undergoing radical prostatectomy, circulating PSA mRNA was detectable in eight patients in basal samples, and in seven of them also in blood specimens collected at the end of surgery, showing an increase in only two patients. In blood samples collected 6 days later, PSA mRNA was dramatically reduced in all patients, but still present in seven of them. In four patients, whose basal samples were negative, PSA mRNA was detectable in samples collected at the end of surgery and three of them were negative after 6 days. In patients who did not receive surgical treatment, a rapid decrease in PSA mRNA was demonstrated in five patients treated with radiotherapy and in one patient undergoing androgen deprivation. No detectable PSA mRNA was found in healthy controls. The levels of PSA mRNA in peripheral blood from patients with prostate carcinoma can be easily measured by this sensitive, quantitative and reliable procedure. This assay is a promising tool for the detection and follow-up of these patients. PMID- 11434387 TI - Serum cardiac troponin I after conventional and minimal invasive coronary artery bypass surgery. AB - We evaluated myocardial release of cardiac troponin I (cTnI) in patients treated with conventional coronary artery bypass grafting (CABG), which employs extracorporeal circulation, and different kinds of minimal invasive coronary artery bypass grafting (MICABG), a surgical technique where the operation is performed without extra-corporeal circulation. Furthermore, we evaluated the usefulness of serum cTnI measurement to detect perioperative myocardial infarction (PMI) after coronary artery bypass surgery. Thirty-one patients were included: sixteen underwent CABG, fifteen underwent different MICABG and five patients had PMI. Blood specimens for cTnI measurements were collected up to 72 hours after opening the graft. Aortic cross-clamping time was a minor determinant of myocardial damage; on the other side, the trauma during surgery correlated with the number of involved arteries and with the manoeuvre employed to obtain heart dislocation, and appeared a more important determinant of myocardial damage. In patients with PMI, the cumulative release of cTnI was higher than in patients free from PMI; however, only after 24-72 hours we observed significant differences in serum cTnI values, because the increased perioperative values of cTnI complicated the interpretation of the myocardial status and a single cut-off could not be used to exclude PMI. PMID- 11434388 TI - Improvement of the quantitative method for glucose determination using hexokinase and glucose 6-phosphate dehydrogenase. AB - This paper has two aims. The first one is to point out the shortcomings of Food and Drug Administration's (FDA's) reference method for the measurement of glucose. We found that the quantity of enzyme used in the method recommended by the FDA was more than the exact quantity needed for accurate measurement. The use of exact quantity of enzyme is important to minimize the negative effects due to impurity and side reactions of enzymes. The second aim is to simulate the coupling enzyme reaction using computer. We have successfully established the exact quantity of enzyme needed in the assay through the computer simulation. The quantity of the enzyme was lesser than the that recommended by FDA, but the reaction ended at the same time as in the FDA method. In addition, optimum conditions and inhibitory effects of various reagents have also been successfully analyzed using computer. In conclusion, we suggest a reference method using computer simulation to determine the exact quantity of the coupling enzyme needed in the assay. PMID- 11434389 TI - Conditions for single-strand conformation polymorphism (SSCP) analysis of BRCA1 gene using an automated electrophoresis unit. AB - The single-strand conformation polymorphism procedure has been applied in routine testing for hereditary diseases and cancer. However, temperature, running time, gel composition, fragment length, etc. can influence its sensitivity. Mutation detection in the clinical setting depends on the development of automated technology, especially for large genes such as the breast cancer gene BRCA1. We analysed DNA samples with BRCA1 mutations in an automated system (GenePhor System; Amersham-Pharmacia Biotech, Uppsala, Sweden). The concentrations of DNA template and PCR primers, the effect of chilling after denaturation, and the temperature and time of the electrophoresis were investigated. All band-shifts were detected by electrophoresis at 5 degrees C for 2 h 15 min. Concentrations of DNA and samples used in the PCR did not affect the SSCP pattern, but chilling the PCR product in ice after denaturation was required. The type and position of mutation in the fragments did not influence the probability of a mobility shift, although SSCP analysis was more sensitive for fragments shorter than 350 bp. This automated SSCP method meets the requirements of fast turnaround and sensitivity and can be readily adapted to the screening of large genes such as BRCA1. PMID- 11434390 TI - Discrepancies between apolipoprotein E phenotyping and genotyping in the elderly. AB - We estimated the frequencies of phenotype (isoelectric focusing; IEF) vs. genotype (PCR/Hhal) discordance in a sample of an aged population (> 65 years). Both phenotype and genotype techniques have been used in the study of apolipoprotein E (apoE) polymorphism in 125 elderly subjects. The discordance between phenotype and genotype was unresolved in 11 (8.8%) of the 125 unrelated subjects studied. We observed a significant association between the presence of the E4 allele and both Alzheimer's disease (chi2 = 13, p < 0.001) and increased cholesterol concentration (Mann Whitney, p < 0.03). These relationships were not affected by the techniques used. Our results indicate that transcriptional modulation and post-transductional modifications in normal ageing and in aged related diseases may explain in part discrepancies between gene analysis and protein characterisation. PMID- 11434391 TI - Markers of bone turnover in postmenopausal women receiving hormone replacement therapy. AB - We investigated the effects of hormone replacement therapy (n = 27) on biochemical markers of bone turnover in a cross-sectional study of 127 postmenopausal women (according to WHO guidelines 18 patients had normal bone mineral density and 109 suffered from bone loss). Urinary excretion of free deoxypyridinoline and C- or N-telopeptide fragments of type I collagen served as bone resorption markers, serum osteocalcin as a bone formation marker. In women with no hormone replacement therapy, only C- and N-telopeptides correlated significantly with the lumbal T-score as an index for bone mineral density. Patients with bone loss receiving hormone replacement therapy exhibited significantly lower C-telopeptide, N-telopeptide and osteocalcin levels than those with no therapy (mean -45%, -43% and -26%, respectively), while deoxypyridinoline showed no significant differences. Among the markers investigated, C- and N-telopeptides seemed to be more reliable to detect therapeutic effects on bone metabolism. We present a preliminary model to evaluate bone turnover and resorption/formation rate. PMID- 11434392 TI - Reference intervals for the markers of proteinuria with a standardised bed-rest collection of urine. AB - Reference intervals for markers of proteinuria or glomerular charge selectivity were measured in 61 healthy female and 61 healthy male individuals. Timed bed rest and daytime collections were used to assess significance of preanalytical variability of results. Bed-rest collections are advisable for research on renal damage, whereas in routine care, robust protein/creatinine ratios work as practical estimates of protein excretion rates, the correlations to excretion rates improving with increasing proteinuria. For glomerular charge selectivity, pancreatic/salivary isoamylase clearance ratio showed lower within-subject biological variation than IgG/IgG4 clearance ratio, allowing more accurate classification into normal and reduced charge selectivity. With our method, the lower 2.5% reference intervals for isoamylase clearance ratio were 1.1 in men and 1.9 in women. PMID- 11434393 TI - The Friedewald formula underestimates LDL cholesterol at low concentrations. AB - Due to recent advances in the treatment of hypercholesterolemia, low density lipoprotein (LDL) cholesterol concentrations below 2.6 mmol/l have become attainable. In general, LDL cholesterol is determined indirectly according to Friedewald. We examined the performance of the Friedewald formula at low concentrations of LDL cholesterol in comparison with a beta-quantification method. We analyzed 176 samples from individuals treated by LDL apheresis with a mean LDL cholesterol concentration of 3.07 mmol/l and found that the Friedewald formula underestimated LDL cholesterol with a bias of -18.5%, -14.5%, -7.3%, and 3.8% at mean LDL cholesterol levels of 1.58, 2.4, 3.49, and 4.67 mmol/l, respectively. Thus, the lower the LDL cholesterol concentration was, the greater the negative bias. We conclude that the Friedewald formula may not be reliable at low LDL cholesterol concentrations produced by LDL apheresis. This finding may also be of relevance to the monitoring of patients being treated with lipid lowering drugs. PMID- 11434394 TI - The white blood cell differential: three methods compared. AB - The analysis of the automated blood cell count is an essential tool in haematological diagnostics. However, in the case of the white blood cell differential the microscopy method, although tedious, often serves as reference. We evaluated the ABX Pentra 120 Retic haematology analyser in comparison to the Coulter STKS haematology system and the microscopy method with respect to accuracy, precision and reliability. We compared 308 samples (239 samples from adults and 69 from children) including patients with oncological diseases. The comparison of the white blood cell differential revealed strong correlations between the results obtained with the ABX Pentra 120 Retic and the microscopy method, the Coulter STKS and the microscopy method and both automated methods (values of paediatric samples in parentheses; neutrophils: rs > or = 0.933 (rs > or = 0.951), lymphocytes: rs > or = 0.907 (rs > or = 0.945), monocytes: rs > or = 0.584 (rs > or = 0.459) and eosinophils: rs > or = 0.963 (rs > or = 0.966)). The analytical performance of automatic analysers for the detection of the morphological "left shift" was determined for all samples in comparison to the microscopical white blood cell differential. The sensitivity, specificity and efficiency depended strongly on the chosen threshold levels and were different for both analysers. The sensitivity for flagging a left shift increased with an increasing proportion of neutrophil bands, metamyelocytes, myelocytes and promyelocytes. Our study suggests that the ABX Pentra 120 Retic haematology analyser, as well as the Coulter STKS haematology system are useful tools for routine analysis in haematology. PMID- 11434395 TI - Plasma brain natriuretic peptide measured by fully-automated immunoassay and by immunoradiometric assay compared. AB - The clinical relevance of measuring plasma brain natriuretic peptide (BNP) is well-known, especially in patients with heart failure. Recently, a new method for measuring BNP, called TRIAGE, has been developed which can be used for point-of care testing of patients with congestive heart failure. The aim of the present study is to compare the analytical performance of this fully-automated method to that of an immunoradiometric assay (IRMA), routinely used to measure BNP. The TRIAGE method is a non-competitive immunofluorometric assay which uses two different binding phases, specific for two different epitopes of the BNP amino acid chain, to form a sandwich with the specific ligand (i.e., BNP). A polyclonal antibody is included in the fluorescent immunoassay reagents which are contained in the assay devices and a monoclonal antibody is immobilized in the detection lane of the assay device. The imprecision of the TRIAGE method was approximately 12% for BNP concentrations in the normal range and about 18% for BNP concentrations above the normal range. The mean reading time of the TRIAGE method was 14.5 +/- 8.6 min. A close linear relationship was found between the BNP values measured with the two methods (TRIAGE = 24.6+0.933 IRMA; r = 0.932, n = 83). The TRIAGE method is indicated for BNP assay in ambulatory and coronary or emergency units, where usually only a few samples (preferentially whole blood samples) must be measured in a short time. The IRMA method should be preferred for pathophysiological studies, requiring the highest degree of precision and sensitivity for simultaneous measurement of several stored plasma samples or tissue extracts. PMID- 11434396 TI - Length of sedimentation reaction in undiluted blood (erythrocyte sedimentation rate): variations with sex and age and reference limits. AB - Although the length of sedimentation reaction in blood (LSRB) (commonly, but improperly called erythrocyte sedimentation rate, ESR) has long been used in clinical laboratories because it is simple and low-cost, its sensitivity and specificity are unsatisfactory. Usually, the values are reported using the Westergren method with sodium citrate-anticoagulated specimens. We used a new procedure, the TEST1, which measures the length of sedimentation reaction in undiluted K3EDTA anticoagulated blood samples following ICSH (International Committee for Standardization in Haematology) recommendations. Samples obtained from 840 reference individuals (430 females and 410 males, mean age 44 and 46.5 years respectively, range 1 to 90 years) were utilised to estimate the reference limits. The subjects, classified by sex, were subdivided into four statistically different age groups to determine the reference limits (2.5th and 97.5th percentiles). Sex difference was statistically significant in two age groups, from 14 to 50 (p < 0.0001) and from 50 to 70 years (p < 0.01). We did not observe significant sex difference within the age bracket from 1 to 14 years and from 70 to 90 years. In both sexes LSRB values increased with age, in significant correlation with fibrinogen concentration (p < 0.0001), and became significantly higher in subjects older than 70 compared to all the younger subjects (p<0.01 in females and p < 0.02 in males). Thus, we defined adequate reference ranges in elderly. PMID- 11434397 TI - Biochemical analysis of the sperm and infertility. AB - The clinical value of biochemical analysis of sperm is still unclear. In this study, we evaluated the potential of several biochemical markers in the seminal plasma (zinc, citrate, acid phosphatase, fructose and neutral alpha-glucosidase) as a screening method in male infertility investigation. PMID- 11434398 TI - Evaluation of a rapid bedside immunochromatographic assay for detection of cardiac troponin I in whole blood. PMID- 11434399 TI - Correct graphic presentation of method comparison data. PMID- 11434400 TI - European symposium on clinical laboratory and in vitro diagnostics industry. PMID- 11434401 TI - A qualitative assessment of the neurological safety of antipsychotic drugs; an analysis of a risperidone database. AB - BACKGROUND: Neurological side effects of antipsychotic agents limit the use of these drugs, and development of newer antipsychotic agents has been focused on a reduced risk of extrapyramidal symptoms (EPS) as well as effective symptom control. METHODS: A qualitative analysis of EPS was performed using Extrapyramidal Symptom Rating Scale (ESRS) data from 11 double-blind risperidone trials. An ESRS factor analysis and maximum changes in ESRS scores were compared for the risperidone, haloperidol, and placebo groups. RESULTS: The factor analysis revealed five factors. Between-group comparisons showed no differences between placebo and 1 to 2 mg/day-risperidone groups. Parkinsonism, tremor, akathisia, and sialorrhea were more likely to occur with haloperidol than with placebo or risperidone at 1 to 6 mg/day. Similar results were noted by maximum changes in ESRS scores. At risperidone doses of more than 8 mg/day, acute EPS severity lay between that of the placebo and haloperidol groups. The severity of tardive dyskinesia was greater in patients receiving placebo than in those receiving either active treatment. CONCLUSIONS: As the results described above were derived from a post hoc analysis of an existing database, conclusions must remain tentative. To provide more definitive answers, EPS assessments in future studies should be refined to more accurately predict the type of EPS expected with a given agent in clinical practice. PMID- 11434402 TI - Autonomic profile under hypnosis as assessed by heart rate variability and spectral analysis. AB - To test the hypothesis of a sympathovagal balance shift towards an enhanced vagal tone under the condition of a standardized hypnosis, we used the assessment of heart rate variability including spectral analysis in 10 healthy subjects (5 female, 5 male, age ranging from 27 to 42 years). We compared the subjects under baseline condition (resting) and under hypnosis, measured on a different day, for a measuring period of five minutes for each condition. The result was reduced total power in the low frequency band (0.01-0.05 Hz) reflecting sympathetic activity. Also, the ratio of low to high frequency power (LFIHF) was reduced when under hypnosis. This reduction was due to a reduced LF component, but only a slightly reduced mid frequency component. With considerable variability, we also found activated mid frequency bands (0.05-0.15Hz) under hypnosis, likely reflecting baroreceptor activity. We conclude that the autonomic status in hypnosis is associated with a change towards reduced low frequency activity, but not necessarily with enhanced high frequency activity. PMID- 11434403 TI - Risk profile of SSrIs in elderly depressive patients with co-morbid physical illness. AB - BACKGROUND: So far, most studies on treatment strategies in elderly depressive patients have included only patients in good physical health, thereby excluding and neglecting somatic co-morbidity, which is very prevalent and relevant in geriatric psychiatry. METHOD: 40 elderly depressive inpatients at the Department of Internal Medicine in Hochzirl who had started on SSRI monotherapy were allocated to this prospective post-marketing surveillance study. A stable medication for their physical illness for at least six months was a prerequisite. A Mini Mental State Exam (MMSE) score of >24 was required for study entry. The four-week study consisted of one baseline and four follow-up examinations, including psychiatric and medical history, as well as ratings for psychopathology and treatment-related adverse events. The antidepressants administered were paroxetine (20 mg/d), citalopram (20 mg/d), fluoxetine (20 mg/d) and sertraline (50 mg/d). Depression was rated using the 21-item Hamilton Depression Scale (HAMD); side effects were evaluated by the UKU Side Effect Rating Scale, and we used the Hillside Akathisia Scale (HAS) to record the incidence of SSRI-induced akathisia. RESULTS AND CONCLUSION: Our results suggest that SSRls are effective and reasonably safe in elderly depressive patients with co-morbid physical illness. Adverse effects are more common, but generally tolerable, than in younger and physically healthy patients. The risk profile of SSRls in this population can be considered favorable. PMID- 11434404 TI - A comparison of citalopram and paroxetine in the treatment of panic disorder: a randomized, single-blind study. AB - BACKGROUND: Serotonin Selective Re-uptake Inhibitors (SSRIs) are the drugs of choice for treating panic disorder (PD). In vitro studies have shown different pharmacodynamic profiles for SSRIs, but their clinical relevance is still unknown. Paroxetine, the SSRI with the strongest serotonergic effect, also shows significant cholinergic and noradrenergic activities. In this class of drugs, citalopram is the most selective for serotonin. We compared these two drugs and their effectiveness and tolerability in a sample of patients with PD in a two month treatment course. METHOD: Fifty-eight patients with PD were randomly assigned to either the paroxetine or the citalopram treatment group in a single blind, randomized design. Each patient was assessed at days 0, 7 and 60 by the Panic Associated Symptoms Scale (PASS), the Sheehan Disability Scale (SDS) and the Fear Questionnaire (FQ). Primary outcome measures were the percentage of patients free of panic attacks, anticipatory anxiety and phobic avoidance in the last week of the trial and the percentage of good responders, as defined by a reduction of at least 50% from baseline of both PASS and SDS global scores at day 60. RESULTS: At day 60, 86% of patients receiving citalopram and 84% of those receiving paroxetine responded well to treatment. No significant differences between the two drugs were found. Both were well tolerated, although sexual side effects and weight gain were frequent. Anticipatory anxiety decreased significantly after the first week of treatment, and no initial worsening in the panic attacks was observed. CONCLUSION: Paroxetine and citalopram show similar anti-panic properties and a good tolerability profile. Our results support evidence that the serotonergic system plays a significant role in the anti-panic properties of these two SSRIs. PMID- 11434405 TI - Quality of partnership in patients with erectile dysfunction after sildenafil treatment. AB - OBJECTIVE: A comprehensive investigation on the quality of partnership of male patients with erectile dysfunction (ED) after treatment with sildenafil vs. untreated patients, as perceived by both the patients and their female partners. METHODS: This report describes an observational, cross-sectional exploratory study comparing ED patients responsive to sildenafil with ED patients prior to therapy. Assessments included the 'International Index of Erectile Function' (IIEF) and the 'Partnerschaftsfragebogen' (PFB), a partnership questionnaire used in German-speaking countries. The comparability of the two study groups was examined using a stepwise logit model. Significant intergroup differences regarding demographics and history were identified and included as confounding variables in the assessment of Quality of Partnership differences using ANCOVA. A regression analysis was performed to determine the association between the mean total IIEF scores and Quality of Partnership measures. RESULTS: 105 patients were included in the study. After adjustment for confounding variables, the groups varied significantly with respect to Quality of Partnership as perceived by men (mean score PFB 61.8 +/- 13.9 vs. 54.4 +/- 15.5; p<0.001) and women (mean score PFB 63.1 +/- 13.6 vs. 57.0 +/- 14.7; p = 0.006). In men, all three PFB subscales (quarreling, tenderness, togetherness) differed significantly between the two study groups. In the female partners, the tenderness and togetherness domains varied significantly. Erectile function correlated highly significantly with tenderness and togetherness in both the male patients and their partners. CONCLUSION: Our data indicate that the Quality of Partnership reported by both the men and their female partners is significantly better in appropriately treated ED patients than in untreated controls. PMID- 11434406 TI - Efficacy and tolerability of hypericum extract WS 5572 versus placebo in mildly to moderately depressed patients. A randomized double-blind multicenter clinical trial. AB - We have investigated the antidepressant efficacy and safety of Hypericum perforatum (St. John's wort) extract WS5572 in a double-blind, placebo-controlled multicenter clinical trial. 72 patients (WS 5572: 37, placebo: 35) with a diagnosis of mild to moderate major depressive disorder (according to DSM-IV criteria) were randomized in 42 days of treatment with either 300 mg WS5572 t.i.d. or placebo. The primary efficacy variable was the change of the 17-item Hamilton Depression Scale (HAMD) total score between baseline and double-blind treatment. The study was conducted with an adaptive interim analysis, which led to early stopping because convincing treatment efficacy could already be demonstrated. Group differences in favor of WS 5572 were descriptively apparent as early as day 7 of randomized treatment and were statistically significant at days 28 (p = 0.011) and day 42 (p < 0.001). Between baseline and treatment end, the HAMD total score decreased from 19.7 +/- 3.4 to 8.9 +/- 4.3 points in the Hypericum group and from 20.1 +/- 2.6 to 14.4 +/- 6.8 points in the placebo group (mean +/- SD). Responder rates were consistently higher in the Hypericum group. Comparable group differences in favor of WS 5572 were also found for von Zerssen's Depression Scale (D-S; self-rating), Clinical Global Impressions (CGI) and a global patient's self-assessment (GPA). Tolerability was very good in both groups, with no adverse drug reactions and no clinically relevant changes in safety parameters. The results indicate that Hypericum extract WS 5572 is an effective and well-tolerated drug for the treatment of mild to moderate major depressive disorder. PMID- 11434407 TI - Mental Health Act assessments: does a community treatment team make a difference? AB - There is an extensive body of research into home treatment for acute mental illness, but this is the first one to examine any association with Mental Health Act Assessments. It examines the question of whether there is any impact of a home treatment team on the numbers and circumstances of Mental Health Act Assessments in its area. The 1983 Mental Health Act created the role of the Approved Social Worker, among whose duties is that of ensuring that any disposal is the least restrictive possible. For this duty to be carried out meaningfully, as wide a range of alternatives as possible needs to be available. The move from institutional to community care for the mentally ill which has been the 'big idea' of recent decades has given rise to intense interest in home treatment for acute mental illness, which can offer a potential alternative at a time of crisis for ASWs and thus patients. This study examined over 200 reports written by ASWs following Mental Health Act Assessments in the two areas, and it was found that the home treatment team appeared to have an impact. It was hypothesised that such a team might have an effect on the sorts people presenting for assessment. It was concluded that area with the team had an absolute and a relative reduction in the numbers of assessments. It was also found that there was a decrease in the likelihood of detention under Section 3 of the Act for people living in the area of the team. Finally it was found that the involvement of the team in an assessment reduced the likelihood of a Section 3 decision even further. Analysis indicated that the home treatment team was able to absorb some of the people, mainly male, who were suffering from a psychotic illness, and who were inclined to re-present on more than one occasion. PMID- 11434408 TI - Against over-interpretation: the understanding of pain amongst Turkish and Kurdish speakers in London. AB - The understanding of experienced pain has recently moved from the biological to the metaphorical. Detailed interviews with twelve Turkish and Kurdish patients in London who had been unsuccessfully investigated medically for chronic pain showed that their understanding reflected local, typically humoural, conceptions of self and body. However there was little to suggest interpretation of the illness as a more specific and grounded idiom for social or political experience. It is suggested that the current vogue for 'interpretation' in medical anthropology and social psychiatry may occasionally be, as Umberto Eco puts it, 'over interpretation'. PMID- 11434409 TI - Quality of life--as defined by schizophrenic patients and psychiatrists. AB - The aim of the study is to explore what schizophrenic patients understand by quality of life and what psychiatrists think quality of life may be like for schizophrenic patients. A survey was carried out among schizophrenic patients and psychiatrists from the whole of Germany. There are similarities and differences between the two: while both schizophrenic patients and psychiatrists recognised the importance of work, social relationships, family and independence for quality of life, patients thought about it more in terms of standard of living and lifestyle, whereas the psychiatrists' concept of quality of life was more illness oriented encompassing the absence of handicaps and disabilities due to the illness and emphasising the importance of appropriate professional help and self help. In order to facilitate the dialogue between psychiatrists and patients it is important to be aware of these differences. PMID- 11434410 TI - Labeling effect of Seishin-bunretsu-byou, the Japanese translation for schizophrenia: an argument for relabeling. AB - Stigma attached to individuals with schizophrenia may be derived from its diagnostic label. Two forms of a questionnaire were distributed to 189 Japanese university students. Each questionnaire contains case vignettes of schizophrenia and major depression. In one form, the students were told at the close of each vignette the diagnostic label assigned to that case (the label group) whereas in another form, they were not done so (the control group). The students were also asked to rate negative image of the case in four items each. The label group was significantly higher than the control group in three of the four negative image items for the vignette of schizophrenia. They did not differ in any of the negative image items for the vignette of depression. This suggests that the label of schizophrenia (the Japanese translation Seishin-bunretsu-byou) has stigmatizing effect. We discussed these findings in the light of the implications of a relabeling, and argued for a change of name. PMID- 11434411 TI - Care of patients with chronic mental illness: comparison of home and half-way house care. AB - Community-based services are being developed as mental health services. The purpose of this study was to examine the effect between home care and half-way house services on patients with schizophrenia. The design was quasi-experimental and used repeated measurements. There were 60 subjects, 37 receiving home care and 23 receiving half-way house care which were affiliated with one general hospital in the south of Taiwan. Of the 60 chronic schizophrenic patients, there were 34 males and 26 females with an average age of 34 years. The Quality of Life Scale was applied to measure each patient four times, at an interval of two months. These follow-up data were analyzed by Generalized Estimating Equation-I (GEE-I) because repeated observations on an individual may be correlated. The result showed that the total QOL in patients receiving home care programs was significantly higher than those receiving half-way house service. The total scores of QOL in patients under half-way house and home care services did not show secular change. The patients under home care program showed a somewhat improvement for a short period of time. However, as time went on, the patients showed no obvious improvement in their life quality. The dimensions of independence and social activity in QOL also showed significantly different between these two groups. The results may provide guides on designing programs and activities for the chronic mental patients. However, the best decision to apply which program for patients need continouous and comprehensive assessment. PMID- 11434412 TI - University students' attitudes towards mental patients and psychiatric treatment. AB - BACKGROUND: Public attitudes towards mental patients and psychiatric treatment are mostly unfavorable. Mental patients tend to conceal their illness or selectively disclose their treatment histories to someone they trust. AIMS: To examine the university students' attitudes towards different information regarding a person labeled as mentally ill. METHODS: We designed seven descriptions of a mentally ill person of varying past and current treatment histories and randomly assigned 308 university undergraduates to read one of the descriptions. Subjects rated their willingness to interact with the hypothetical case using a 19-item social distance scale. RESULTS: Greater social distance was associated with non-medical field of study, no previous contact with the mentally ill and female gender. Subjects without previous contact with mentally ill individuals kept greater distance from a discharged mental patient receiving psychiatric care than a mental patient who did not require medications nor psychiatric follow-up. In contrast, respondents who had previous contact with the mentally ill were more willing to interact with a discharged mental patient receiving psychiatric care comparing with a person labeled as mentally ill alone and a mental patient who had never been admitted to hospital. CONCLUSIONS: Implications of our findings in clinical management and designing stigma-reducing programme are discussed. PMID- 11434413 TI - Initiating care for untreated schizophrenia patients and results of one year follow-up. AB - A number of schizophrenia patients live untreated in the community in the developing countries. There is little recorded experience of how such patients would respond to treatment after years of untreated illness. A cohort of 72 never treated chronic schizophrenia patents in Chennai, India were directed to attend a health facility. A substantial proportion of them (68%) came for treatment. Unemployed status of male patients, living in a joint family setting and families initially unaware of the psychiatric nature of the problem were the factors that related to failure to seek treatment. Patient's sex, age, education, marital status, economic status, age at onset and duration of illness, degree of disability and clinical symptoms (except self-neglect) were not related to taking treatment. Those who attended were treated with typical antipsychotic drugs and followed up for one year. Evaluation was done using the Present State Examination and Psychiatric History and Sociodemographic Schedule and Disability Assessment Schedule. The clinical outcome was good (Best Remission) in 29%. There was no impairment in social functioning in 35% and 51% has no impairment in occupational functioning at the end of one year. PMID- 11434414 TI - Biological differences in depression and anxiety across races and ethnic groups. AB - A growing number of studies clearly indicate the importance of race and ethnicity in the psychopharmacologic management of depression and anxiety disorders. The data highlight important pharmacokinetic, pharmacodynamic, and pharmacogenetic ethnic differences that may have profound implications for the efficacy and safety of psychotropic therapies. General treatment considerations based on these differences include greater attention to adverse event profiles, the possibility of improved clinical response at any given dose, and the potential need for lower starting doses and slower increases in dosage. Continued research in this area is clinically important as patients with increasingly divergent ethnic and cultural backgrounds seek treatment for a range of depressive and anxiety disorders. PMID- 11434415 TI - Cultural variations in the clinical presentation of depression and anxiety: implications for diagnosis and treatment. AB - This article reviews cultural variations in the clinical presentation of depression and anxiety. Culture-specific symptoms may lead to underrecognition or misidentification of psychological distress. Contrary to the claim that non Westerners are prone to somatize their distress, recent research confirms that somatization is ubiquitous. Somatic symptoms serve as cultural idioms of distress in many ethnocultural groups and, if misinterpreted by the clinician, may lead to unnecessary diagnostic procedures or inappropriate treatment. Clinicians must learn to decode the meaning of somatic and dissociative symptoms, which are not simply indices of disease or disorder but part of a language of distress with interpersonal and wider social meanings. Implications of these findings for the recognition and treatment of depressive disorders among culturally diverse populations in primary care and mental health settings are discussed. PMID- 11434416 TI - Prescribing patterns for depression and anxiety worldwide. AB - Psychiatric disorders, particularly depression and anxiety, present commonly in primary care worldwide. However, significant variation is seen between countries in the prevalence of these disorders. Differing levels of awareness and recognition due to cultural issues may be important contributing factors in this variation. Overall, roughly half of psychiatric cases presenting in primary care go unrecognized, and one third of cases recognized are misdiagnosed. Although treatment is offered in nearly 60% of recognized cases, it is seldom in the context of a specific diagnosis and is only appropriate in about 5% of cases overall. This article explores the transcultural variation in the recognition, diagnosis, and treatment of depression and anxiety disorders using data from the World Health Organization study on Psychological Problems in General Health Care. Patterns observed in Western societies are compared with those seen in the rest of the world and are discussed with a view to educational needs for primary care physicians. PMID- 11434417 TI - Mental health care in Japan: recognition and treatment of depression and anxiety disorders. AB - Despite the apparent high availability of psychiatric services in Japan, the quality and type of care have historically been lower than that provided in Western society. However, Japanese psychiatry is undergoing a period of transition. Recent changes in the provision of health care, the adoption of internationally standardized diagnostic criteria, the availability of newer antidepressant drugs such as selective serotonin reuptake inhibitors, and other social and economic factors mean that psychiatric services are improving at various levels. PMID- 11434418 TI - Epidemiology, burden, and disability in depression and anxiety. AB - Studies of the prevalence of depression and anxiety disorders have shown that there is a high prevalence of comorbidity of these 2 disorders. The resulting disability and burden affect not only the individual in terms of decreased productivity, but the level of health care utilization is also increased. The objective of this article is to look at the epidemiology, disability, and global burden of depression and anxiety across the different nations of the world. This article will concentrate on the results from the Cross-National Collaborative Group. The transcultural trends in prevalence and disability presented here must be viewed in the light of the limitations of the study, such as methodology and population sampling, uniformity in the method of clinical assessment, and the collection and processing of data. New studies of depression and anxiety among different cultures are currently in progress in the form of the European Study of Epidemiology on Mental Disorders (ESEMeD), which is closely linked to the World Health Organization (WHO) World Mental Health 2000 initiative. The methodology for ESEMeD is similar to that of the WHO World Mental Health 2000 study, which will facilitate comparisons between the results for Europe and the rest of the world. Results of these studies are awaited with anticipation. PMID- 11434419 TI - Consensus statement on transcultural issues in depression and anxiety from the International Consensus Group on Depression and Anxiety. AB - OBJECTIVE: To provide primary care physicians with a better understanding of transcultural issues in depression and anxiety. PARTICIPANTS: The 4 members of the International Consensus Group on Depression and Anxiety were James C. Ballenger (chair), Jonathan R. T. Davidson, Yves Lecrubier, and David J. Nutt. Five faculty invited by the chair also participated: Laurence J. Kirmayer, Jean Pierre Lepine, Keh-Ming Lin, Osamu Tajima, and Yutaka Ono. EVIDENCE: The consensus statement is based on the 5 review articles that are published in this supplement and the scientific literature relevant to the issues reviewed in these articles. CONSENSUS PROCESS: Group meetings were held over a 2-day period. On day 1, the group discussed the review articles, and the chair identified key issues for further debate. On day 2, the group discussed these issues to arrive at a consensus view. After the group meetings, the consensus statement was drafted by the chair and approved by all attendees. CONCLUSION: The consensus statement underlines the prevalence of depression and anxiety disorders across all cultures and nations while recognizing that cultural differences exist in symptom presentation and prevalence estimates. In all countries, the recognition of depression by clinicians in the primary care setting is low (generally less than 50%), and the consensus group recommends a 2-step process to aid the recognition and diagnosis of depression. In line with the low recognition of depression and anxiety disorders is the finding that only a small proportion of patients with depression or anxiety are receiving appropriate treatments for their condition. Biological diversity across ethnic groups may account for the differential sensitivity of some groups to psychotropic medication, but this area requires further investigation. PMID- 11434420 TI - The profiles of Kawasaki disease in China. AB - Kawasaki disease in China was described for the first time in 1976 in Taiwan, and in 1978 in mainland, respectively. Questionnaire surveys had been conducted in both of the area of China and showed that the Kawasaki disease patients increased year by year. No data on incidence rates were available for these surveys because the problem of representativeness. However, it showed that there were many similar characteristics of Kawasaki disease in China comparing with those in Japan. Although a series of infectious agents were suspected, the etiology of Kawasaki disease remained unclear. High dose of gamma globulin treatment was also adopted commonly in China. PMID- 11434421 TI - Possible risk reduction in esophageal cancer associated with MPO -463 A allele. AB - Myeloperoxidase (MPO), an enzyme found in lysosomes of phagocytes, causes hydroxy radicals linked to DNA damage and activation of smoking related carcinogens. A 463 G/A polymorphism in the promoter region of the MPO gene results in reduced gene expression, which would imply lower susceptibility of esophageal cancer in mutant carriers. We conducted case-control study to test this hypothesis. Cases were 91 patients with esophageal cancer and controls were 241 non-cancer outpatients. MPO genotypes were examined by PCR-RFLP. The allele frequency for MPO -463A was found to be 8.2% for cases and 10.5% for controls. The age, sex, smoking and drinking status adjusted odds ratio for all subjects for MPO -463 GG/GA as compared to the AA was 0.61 (95% CI: 0.28-1.32). The adjusted odds ratio for the GG/GA genotype was significantly low (0.15; 0.03-0.76, P=0.022) for those aged 61 years or older who had a significantly higher odds ratio for smoking than younger subjects. No difference was observed in disease risk when prevalent and incident cases were compared. Although there are limitations for interpretation of this study because of prevalent case-control study and partial statistical significance, these results suggest that MPO -463 A allele reduce the risk of esophageal cancer. PMID- 11434422 TI - Angiotensinogen gene variation and hypertension in a cohort study in Japanese. AB - Many recent case-control studies have suggested a significant relationship between M235T (the substitution of threonine for methionine at position 235 codon) polymorphism of the angiotensinogen (AGT) gene and hypertension. To investigate whether the M235T polymorphism of AGT gene affects the incidence of hypertension, a retrospective cohort study was performed among Japanese workers. The subjects were Japanese workers at an occupational site in Shimane Prefecture in Japan. The baseline data were set at the received regular health examination in 1992, and a retrospective cohort study was performed for analyzing the incidence of hypertension in 1998. The rates of M235M (MM), M235T (MT) and T235T (TT) genotypes were 4%, 32% and 64%, respectively. The relative risks of MT and TT against MM for the incidence of hypertension by single variance analysis were 1.47 [95% confidence interval (CI) 0.50 - 4.33] and 1.35 (95% CI 0.47 - 3.90), respectively. The relative risks of MT and TT against MM for the incidence of hypertension, adjusted for sex, age, body mass index, fasting glucose and cigarette smoking, drinking and exercise in 1992, were 1.49 (95% CI 0.49 - 4.53) and 1.25 (95% CI 0.42 3.74), respectively. The data from this study suggest that the M235T polymorphism of AGT gene has a weak role in the manifestation of hypertension. Further comprehensive studies are needed to resolve this issue. PMID- 11434423 TI - Smoking habit and interleukin lB C-31T polymorphism. AB - Recent studies suggest that smoking habit may relate to genetic traits. This study examines the association with a polymorphism (C-31T) of interleukin 1B (IL 1B), which encodes IL-1beta, a multifunctional pro-inflammatory cytokine. Since the T allele makes a TATA box, the allele is thought to be responsible for a higher potency of IL-1B expression, indicating that individuals harboring the T allele are prone to inflammation. The study subjects were two different populations; 241 non-cancer outpatients (118 males and 123 females) at a cancer hospital and 462 examinees (127 males and 335 females) of a health checkup program provided by a local government. Current smokers were 36.4% for the male outpatients, 9.8% for the female outpatients, 38.6% for the male examinees, and 5.6% for the female examinees. The sex-age-adjusted odds ratios of current smokers were calculated for the genotypes with the T allele relative to the CC genotype by an unconditional logistic model. The estimate was 0.45 (95% confidence interval, 0.21-0.97) for the outpatients, and 0.83 (0.42-1.61) for the examinees. Although not significant for the examinees, the observed associations suggest that this polymorphism may influence smoking behavior through an inflammatory response of the respiratory tract to cigarette smoke. PMID- 11434424 TI - Tinea capitis infection in school children of Nepal. AB - From among 428 Nepalese schoolchildren hair samples of 102 children with clinical features of tinea capitis, obtained by the sterile hairbrush method, were examined by mycological techniques. Age varied between 4-16 years. Itching was experienced by 96.1% sample subjects and hair loss by 32.4%. Of the 102, 11 (10.8%) were positive for Trichophyton violaceum (TV), 6 being from urban areas, the rest from rural areas. Amongst the 11 patients, 7 (63.6%) were girls and rest boys. Statistical associations were observed between the place of haircut and isolation of the organism (chi2 = 15.2, p <0.01). Statistical association was also present between frequency of bathing and isolation of organism. Sharing of combs was associated with the culture-positive subjects. The prevalence of tinea capitis in the urban and rural children was 2.3% and 3.0%, respectively. The only isolated organism was TV. An association of the isolation of TV was found with risk factors such as family members, sharing of combs, frequency of bathing with the organism. Hair loss was more common in the urban children. Discouragement of sharing combs, increased frequency of hair washing, and use of uncontaminated hair cutting instruments are recommended. PMID- 11434425 TI - Apolipoprotein E genetic polymorphism and stroke subtypes in a Bangladeshi hospital-based study. AB - The association between apolipoprotein E (apoE) genetic polymorphism and stroke has not been concordant in different racial populations. We investigated the association between apoE genotypes and stroke subtypes by a case-control study in Bangladesh for the first time among south Asian countries. First-ever-stroke patients (n=227; cerebral infarction, n=147, cerebral hemorrhage, n=80) and 190 controls were recruited from a hospital in Dhaka, Bangladesh. The diagnosis of stroke was based on CT and clinical findings. Cerebral infarction was classified anatomically into cortical and penetrating region. Infarction in the cortical region was further categorized etiologically into thrombosis and embolism. Cerebral hemorrhage was considered as a whole in all analyses. ApoE genotypes were determined by restriction fragment length polymorphism. In the multivariate conditional logistic regression analysis adjusted for potential confounders both the epsilon3/epsilon4 genotype and epsilon4 carrier conferred an approximately 3 fold increased risk for cerebral thrombosis in the cortical artery region (OR 3.5, 95% CI 1.2 to 10.4 and OR 3.1, 95% Cl 1.1 to 9.0, respectively) compared with epsilon3/epsilon3 genotype. However, when the analysis was restricted to the elderly (>60 years), epsilon 2 carrier was associated with a risk of hemorrhagic stroke (OR 19.2, 95% CI 1.3 to 295.2). Our study suggested that both apoE epsilon3/epsilon4 genotype and epsilon4 carriers were risk factors for cerebral thrombosis in cortical artery region, whereas epsilon 2 carrier was a risk factor for hemorrhagic stroke in the elderly. PMID- 11434426 TI - Accidents associated with bathing in home care services for the aged in Japan. AB - OBJECTIVES: To reveal what kinds of accidents happen associated with bathing in home care services for the aged in Japan. METHODS: The study was cross-sectional in design. In November 1999, a postal questionnaire survey was conducted of 828 councils selected from the list of the National Council of Social Welfare of Japan by a systematic sampling method (extraction rate was 25%). The main outcome measures were characteristics of cases of accidents associated with bathing service for the aged, including patient age, sex, time of occurrence, symptoms, and results of accidents. RESULTS: Replies were received from 683 (82%) councils. Of the councils that replied, 430 (63%) reported providing bathing service for the aged. Of these 430 councils, 108 (25%) have experienced accidents, and 130 cases were analyzed. Affected patients had a mean age of 80.7 years (SD: 9.0 years), and 71 were females (55%). Sixty-two (48%) had symptoms of a disease or had accidents after bathing, and 42 (32%) presented with loss of consciousness. In results of accidents, 14 (11%) were reported to have died. CONCLUSION: This study revealed that there were not a few accidents associated with bathing in home care services for the aged in Japan, the most frequent symptom was loss of consciousness, most accidents occurred after bathing, and that some patients died as a result of bathing provided by home care services. PMID- 11434427 TI - Molecular virology of hepatitis C virus. AB - Hepatitis C virus (HCV), discovered in 1989, is the major causative agent of parenteral non-A, non-B hepatitis worldwide. Following the development of a method of diagnosing HCV infection, it became apparent that HCV frequently causes chronic hepatitis. Persistent infection with HCV is implicated in liver cirrhosis and hepatocellular carcinoma. Current worldwide estimations suggest that more than 170 million people have been infected with HCV, an enveloped positive single stranded RNA (9.6-kilobases) virus belonging to the Flaviviridae. The HCV genome shows remarkable sequence variation, especially in the hypervariable region 1 of the E2 protein-encoding region, and globally, HCV appears to be distributed with more than 30 genotypes. Complicated "quasispecies" and frequent mutations of viral genomes have also emerged. The HCV genome encodes a large polyprotein precursor of about 3,000 amino acid residues, and this precursor protein is cleaved by the host and viral proteinases to generate at least 10 proteins in the following order: NH2-core-envelope (E1)-E2-p7-nonstructural protein 2 (NS2)-NS3 NS4A-NS4B-NS5A-NS5B-COOH. These viral proteins not only function in viral replication but also affect a variety of cellular functions. Although several explanations have been proposed, the mechanisms of HCV infection and replication in targeted cells, the mechanism of persistent viral infection, and the pathogenesis of hepatic diseases (hepatitis or hepatocellular carcinoma) are all poorly understood. A major reason why these mechanisms remain unclear is the lack of a good experimental HCV replication system. Although several classical trials using cultured cells have been reported, several new, more promising experimental strategies (generations of infectious cDNA clone, replicon, animal models, etc.) are currently being designed and tested, in order to resolve these problems. In addition, new therapies for chronic hepatitis have also been developed. The enormous body of information collected thus far in the field of HCV research is summarized below, and an overview of the current status of HCV molecular virology of HCV is provided. PMID- 11434428 TI - Nephrin is an important component of the barrier system in the testis. AB - Nephrin, a gene product of the congenital nephrotic syndrome of the Finnish type (NPHS1), is a 1242-residue putative transmembrane protein of the immunoglobulin family of cell adhesion molecules. The expression of this gene is localized in rat and human glomerular epithelial cells. Here we report the expression of nephrin in various tissues other than the kidneys in mice. The expression of nephrin mRNA in various tissues of mice, including the kidneys, testes, spleen, thymus and brain, were first investigated by the RT-PCR method, and it was shown that a high level of nephrin mRNA could be detected in the testes of mice 1-6 weeks old. In situ hybridization revealed the expression of the nephrin gene in the Sertoli cells. Additionally, immunofluorescent staining studies indicated that nephrin was colocalized with anchoring protein ZO-1 in the mouse testis. From these results, it is inferred that nephrin is an important component of the barrier system in testes. PMID- 11434429 TI - Genetic alteration of penicillin non-susceptible Streptococcus pneumoniae observed throughout recurrence of acute otitis media detected by amplified fragment length polymorphism analysis. AB - The prevalence of penicillin non-susceptible Streptococcus pneumoniae (PNSSP) is increasing among isolates from acute otitis media (AOM). Repeated episodes of antibiotic exposure are a well-known risk factor for the isolation of PNSSP although otitis-prone or recurrent AOM cases frequently require repeated courses of antibiotic treatment. In order to evaluate the chronological alteration of S. pneumoniae during recurrences of AOM, strains of S. pneumoniae were isolated from 11 patients, each of whom had experienced 2-4 episodes of AOM, were examined. Every bacterial specimen obtained from a single episode of recurrent AOM was examined by PCR-based penicillin-binding protein (PBP) assay, serotyping, and amplified fragment length polymorphism (AFLP), then compared to other samples from the same case. Two cases (18.2%) showed strain diversity during repeated antibiotic treatments by serotyping or PBP-assay. By AFLP analysis, 6 cases (54.5%) demonstrated heterogeneous strains during recurrent AOM. Clonal survivors of previous episodes of AOM were not always the cause of subsequent episodes of AOM, even in otitis-prone cases. PMID- 11434430 TI - Human identification from forensic materials by amplification of a human-specific sequence in the myoglobin gene. AB - We developed a method for human identification of forensic biological materials by PCR-based detection of a human-specific sequence in exon 3 of the myoglobin gene. This human-specific DNA sequence was deduced from differences in the amino acid sequences of myoglobins between humans and other animal species. The new method enabled amplification of the target DNA fragment from 30 samples of human DNA, and the amplified sequences were identical with that already reported. Using this method, we were able to distinguish human samples from those of 21 kinds of animals: the crab-eating monkey, horse, cow, sheep, goat, pig, wild boar, dog, raccoon dog, cat, rabbit, guinea pig, hamster, rat, mouse, whale, chicken, pigeon, turtle, frog, and tuna. However, we were unable to distinguish between human and gorilla samples. This method enabled us to detect the target sequence from 25 pg of human DNA, and the target DNA fragment from blood stored at 37 degrees C for 6 months, and from bloodstains heated at 150 degrees C for 4 h or stored at room temperature for 26 years. Herein we also report a practical application of the method for human identification of a bone fragment. PMID- 11434431 TI - Time course changes of nerve conduction velocity in idiopathic carpal tunnel syndrome after endoscopic surgery. AB - A total of 49 patients (71 hands) underwent endoscopic carpal tunnel release according to the method of Okutsu accompanied by electrophysiological studies performed for idiopathic carpal tunnel syndrome at our department between August 1993 and May 1998. Among them, 41 patients (55 hands) who were followed-up for 12 months or more postoperatively were studied. The clinical outcome was favorable in 50 out of 55 hands (90.9%). In the 5 hands showing poor surgical results, distal motor latency or sensory nerve conduction velocity was not recordable before surgery and had not improved at 1 month postoperatively. In the present study, it is highly likely that the long-term prognosis will be poor when distal motor latency or sensory nerve conduction velocity is not recordable at 1 month after surgery as well as preoperatively. It is possible to identify patients with a poor outcome by performing electrophysiological studies at 1 month after surgery. PMID- 11434432 TI - My personal experience with speech therapy for stuttering at the Stuttering Center of Western Pennsylvania. PMID- 11434433 TI - Computer simulation of particle overlap in fiber count samples. AB - Fibrous aerosols are of great importance to industrial hygienists because of the severe health risks that may be associated with inhaling such particles. Previous studies on measurement error due to overloading of fibers and nonfibrous particles on the collected sample indicate that a 100-1300 fiber/mm2 filter area is the best filter loading density to reduce bias in fiber counts. The present study investigated the upper fiber and particle concentration limits for reliable counting and identification and the possibility of a procedure for correcting observed fiber counts to account for fiber masking due to overlapping particles or fibers. A computer-generated grid was used to simulate the light microscope graticule field. The resolution of 2000 x 2000 was found to accurately represent the shape of the fibers and nonfibrous particles. Bivariate lognormal distributions were used to describe the length and width distributions of the fibers. The capability of distinguishing particle-overlapped fibers (defined as the resolution index), the coverage of the graticule field, the filter surface loading density, size distributions of fibers and particles, and the fiber-to particle concentration ratio were the primary parameters in this study. The counting efficiency was found to consistently decrease with increasing filter surface loading density and decreasing resolution index. The recommended upper limit of filter surface fiber density depended not only on the number concentration ratio but also on the filter surface loading densities and size distributions of fibers and particles. The advantage of using a thoracic preseparator on counting efficiency was calculated and found to improve counting efficiency significantly when the count median diameter of nonfibrous particles was close to or larger than the thoracic 50% cutoff point of 10 microm. PMID- 11434434 TI - Pressure drop and service life predictions for respirator canisters. AB - Respirator canister designers and manufacturers have a responsibility to provide canisters with increasingly higher capacity and longer service life due to rising safety standards in the workplace. Optimizing the service life of conventional canisters is not about simply increasing the depth of adsorbent present in the canister because this will lead to an increased resistance to airflow, and a greater physiological burden on the user. To optimize canister design it is necessary to establish how the physical dimensions of the adsorbent bed affect breathing resistance and adsorption performance. The study presented in this article investigated both the pressure drop and cyanogen chloride chemisorption performance of a granular activated carbon adsorbent. Results obtained show relationships that correlate adsorption performance and pressure drop to the quantity of adsorbent, the linear flow velocity, and, where applicable, the challenge vapor concentration. Altering the linear velocity by increasing cross sectional surface area has a more beneficial impact on canister performance than altering the adsorbent bed depth. The application of these relationships to the design of canisters is demonstrated. PMID- 11434435 TI - Development of an integrated microanalytical system for analysis of lead in saliva and linkage to a physiologically based pharmacokinetic model describing lead saliva secretion. AB - There is a need to develop reliable portable analytical systems for biomonitoring lead (Pb) in noninvasively collected saliva samples. In addition, appropriate pharmacokinetic analyses are used to quantitate systemic dosimetry based on the saliva Pb concentrations. A portable microfluidics/electrochemical device was developed for the rapid analysis of Pb based on square wave anodic stripping voltammetry, in which a saliva sample flows over an electrode surface, Pb2+ is chemically reduced and accumulated, and the electric potential of the electrode scanned. The system demonstrates a good linear response over a broad Pb concentration range (1-2000 ppb). To evaluate the relationship between saliva and blood Pb, rats were treated with single oral doses ranging from 20 to 500 mg Pb/kg of body weight, and 24 hours later were administered pilocarpine, a muscarinic agonist to induce salivation. To correlate saliva levels with internal dose, blood and saliva were collected and quantitated for Pb by inductively coupled plasma-mass spectrometry (ICP-MS) and by the microanalytical system. The quantitation with the microanalytical system was slightly less (approximately 75 85%) than with ICP-MS; however, the response was linear, with concentration suggesting that it can be used for the quantitation of salivary Pb. To facilitate modeling, a physiologically based pharmacokinetic (PBPK) model for Pb was modified to incorporate a salivary gland compartment. The model was capable of predicting blood and saliva Pb concentration based on a limited data set. These results are encouraging, suggesting that once fully developed the microanalytical system coupled with PBPK modeling can be used as important tools for real-time biomonitoring of Pb for both occupational and environmental exposures. PMID- 11434436 TI - Numerical simulation of airflow around a variable volume/constant face velocity fume cupboard. AB - Three-dimensional models of a variable volume/constant face velocity fume hood were developed and numerically solved to study the effect of the sash opening and rated face velocity on the flow patterns, especially in the exposure area of a person standing in front of the fume hood. These flow details are important in assessing the containment performance of the fume hood. For this study, fully turbulent flow was assumed to enter through the front hood window and exit through the exhaust duct. The finite-difference, control-volume technique, and standard k-epsilon models were employed and solved together with the equations for the conservation of mass and momentum using FLUENT/UNS version 4.2 to obtain the flow solutions. Unobstructed flow cases, as well as the simplified two dimensional scenarios, also were investigated for comparison. Results indicated that there is no occurrence of recirculation leading to concentration buildup within the region between the person and fume hood entrance. Also, in conformance with other studies, the recirculating flow volume inside the hood decreased with increasing sash openings. PMID- 11434437 TI - Test methods for evaluating the filtration and particulate emission characteristics of vacuum cleaners. AB - The overall filtration efficiency of a vacuum cleaner traditionally has been tested by placing the vacuum cleaner in a test chamber and measuring aerosol concentrations at the chamber inlet and outlet. The chamber test method was refined and validated in this study. However, this chamber test method shows an overall filtration efficiency of close to 100% for most of the industrial vacuum cleaners and for most of the newly developed household vacuum cleaners of midprice range or higher because all these vacuum cleaners have a high-efficiency particulate air (HEPA) or other highly efficient filter installed at the exhaust. A new test method was therefore developed through which the vacuum cleaner was probed in various internal locations so that the collection efficiency of the individual components could be determined. For example, the aerosol concentration upstream of the final HEPA filter can thus be measured, which permits one to estimate the life expectancy of this expensive component. The probed testing method is particularly suitable for field evaluations of vacuum cleaners because it uses compact, battery-operated optical particle size spectrometers with internal data storage. Both chamber and probed tests gave the same results for the aerosol filtration efficiency. The probed testing method, however, also gives information on the performance of the individual components in a vacuum cleaner. It also can be used to determine the dust pickup efficiency and the degree of reaerosolization of particles collected in the vacuum cleaner. PMID- 11434438 TI - Inhalable dust exposures, tasks, and use of ventilation in small woodworking shops: a pilot study. AB - Measures of workers' inhalable dust exposures, tasks, and ventilation use were made in five small woodworking shops prior to the start of an intervention effectiveness study aimed at lowering personal wood dust exposures. The data were used to (1) design a sampling protocol for an intervention success measure, (2) identify targets for intervention among the tasks and activities responsible for high dust levels, and (3) develop shop-level measures as tools for tailoring intervention activities. Geometric mean dust concentrations ranged from 1.6 to 9.9 mg/m3 in the five shops, with the highest levels occurring in a cabinet shop. All shops had centralized dust collection systems and workers generally used dust control on stationary tools (60-100% of the time) when it was available. Sanding with both stationary and handheld powered tools, cleaning with methods that can disperse dust (e.g., brushes, compressed air), and miscellaneous tasks were all responsible for significant personal exposures. The positive association between miscellaneous tasks and exposures probably reflects the high background levels generated by nearby processes. Sanding with both stationary tools and handheld powered tools represents the most significant influence on personal exposures in small woodworking shops. The authors conclude that pilot studies are useful tools for designing occupational health and safety intervention effectiveness studies. PMID- 11434439 TI - Emissions of chemical compounds and bioaerosols during the secondary treatment of paper mill effluents. AB - This study identified and quantified the main chemical compounds--the substances responsible for the disagreeable odors--and the bioaerosols emitted during the biological treatment of paper mill effluents. It also identified the characteristics of the process that effects the generation or diffusion of these substances. All treatment stages were evaluated. Measuring sites were located as closely as possible to the potential emission sources. Measurements were taken in the summer in 11 paper mills during a 2- to 3-day period in each mill. Chemical compounds were evaluated by direct-reading instruments; bioaerosols were sampled by impaction and counted. Sulfur compounds, emitted into the air when the effluent or the sludge is stirred, had the highest concentrations; their presence was attributable to such things as kraft-type paper pulp. Next in concentration were the carbon and nitrogen oxides, ammonia, and some organic acids, produced by the action of microorganisms. These acids are found mainly in the sludge environment. Terpenes, which come from wood, are present at various locations in paper mills. Odor perception thresholds for most of these substances are much lower than those established to protect the health of workers. Significant concentrations of total bacteria, total molds, and endotoxins were measured at several sites. Gram-negative bacteria were high at only one site, whereas the mold Aspergillus fumigatus was occasionally present at low concentration. No actinomycetes bacteria were detected. The highest concentrations were measured where there was water or dust aerosolization. Emissions are therefore controlled by controlling the operations that lead to the dispersion of water and particles into the air. PMID- 11434440 TI - An evaluation of short-term exposures to metalworking fluids in small machine shops. AB - In a study of 23 small machining shops using metalworking fluids (MWFs), real time air monitoring using an aerosol photometer was performed to investigate the temporal nature of the exposure and to examine the relationship between the instrumental measurements and traditional sampling methods. Time-weighted averages were calculated from the aerosol photometer data and the results were compared to collocated thoracic and 37-mm closed face cassette samplers. The filter samples were analyzed for total mass and the solvent extractable fraction. Depending on the averaging period used, short-term MWF concentrations exceeded 2.0 mg/m3 in 13 to 39% of the plants studied. High short-term exposures were as likely to be found in plants with average concentrations below 0.4 mg/m3 (thoracic-gravimetric) as those above. Regression analyses indicated that the aerosol photometer most closely matched the data obtained from the thoracic fraction of the total mass. In general, the aerosol photometer overestimated the levels determined using the thoracic cyclone and filter, especially when measuring concentrations of water-based fluids. Use of a calibration factor of 0.7 for straight oils or 0.5 for water-based fluids may assist in the interpretation of aerosol photometer measurements if field calibration data are not readily available. Several approaches to determining the calibration factor from field data were evaluated; more complex calibration techniques improved the accuracy of the measurements. PMID- 11434441 TI - Technology-enhanced learning/distance education: market survey of occupational health and safety professionals. AB - A market survey of occupational health and safety professionals was performed to assess their interest in course work offered through distance education, using technology-enhanced learning methods such as the Internet or CD-ROM. A random sample of 800 active and student members of the American Industrial Hygiene Association, the American Association of Occupational Health Nurses, and the American Society of Safety Engineers from the eight-state Midwest region were queried through a mail survey. Respondents expressed a high likeliness (87.4%) to participate in distance education opportunities for the purposes of continuing education and academic degree. The areas of study interest selected most often were occupational health (73%), injury prevention and control (60%), and industrial hygiene (53%). More than three-quarters of respondents (79%) said that an on-campus component was not important to their learning experience. The majority of respondents (68%) indicated that they were reimbursed for the cost of education with significant differences identified by association. Occupational health and safety professionals are interested in distance education using technology-enhanced learning (TEL) methodologies for meeting their educational needs. TEL/distance education, built on a tested educational approach, should be implemented and outcomes shared to increase the body of knowledge regarding these teaching strategies as they pertain to occupational health and safety professionals. PMID- 11434442 TI - Metalworking fluid exposures in small machine shops: an overview. AB - Sampling was conducted in 79 small machine shops to assess airborne exposures to metalworking fluids (MWFs). Measured exposures were compared with data from the literature and exposure criteria currently recommended by the National Institute for Occupational Safety and Health and the Occupational Safety and Health Administration MWF Standards Advisory Committee. Sixty-two percent of 942 personal samples collected were less than the recommended exposure limit (REL) of 0.50 mg/m3 for total particulate. However, at least 1 sample exceeded the REL in 61 of the 79 facilities studied; 100% of the samples collected in 10 shops were greater than the REL. Similar trends were found for thoracic particulate exposures where 75% of 238 samples were below the thoracic particulate REL of 0.40 mg/m3. The ratio between thoracic and total particulate for 238 paired samples was 0.55 (r2=0.73). Workers exposed to straight fluids had the highest exposures (GM=0.67 mg/m3) when compared with workers exposed to other classes of MWFs. The highest exposures were measured for grinding and hobbing (GM=0.67 and 0.60 mg/m3, respectively). Measurements using personal impactors indicated that particle size distributions of MWF aerosols had an average mass median aerodynamic diameter of 5.3 microm. Straight oils and soluble fluids tended to be associated with larger particles than were other fluid types; grinding and turning produced the largest particles, whereas hobbing resulted in the smallest. In general, exposures were similar in magnitude and particle size to those previously reported in large automotive plants. Therefore, workers in these small shops may have risks of adverse health effects similar to those demonstrated in the automotive industry. PMID- 11434443 TI - Evaluation of the effectiveness of following up laboratory reports of elevated blood leads in adults. AB - The usefulness of a statewide laboratory-based blood lead surveillance system to initiate workplace enforcement inspections was studied. In particular, the effectiveness of inspection of companies in which at least one worker had a blood lead level (BLL) 30-39 microg/dL but no greater than 39 microg/dL was considered. The surveillance system identified all individuals who had been tested for blood lead. Adults with BLLs > or =30 microg/dL were interviewed. Companies where lead exposures occurred were identified and enforcement inspections performed there. Companies were grouped into four categories: one or more employees with BLL > or =50 microg/dL; one or more employees with highest BLL of 40-49 microg/dL; one or more employees with highest BLL of 30-39 microg/dL; and companies that used lead but from which no blood lead reports were received. The number of citations and amounts of penalties, and BLLs before and after, were compared among the three blood lead groups and with a control group of no lead-using companies. A cost benefit analysis was performed. Citations and penalties did not differ among the three blood lead groups but were markedly increased compared with the lead using but no blood lead group. Violations of specific lead standard components were similar among the three blood lead groups. Blood lead companies had increased citations and penalties as compared with nonusing lead control companies. No significant decrease in blood leads was seen postinspection. Total cost to identify lead-exposed workers at problem work sites was $125 ($53-$459) per lead exposed worker. Followup of companies identified through the surveillance system was an effective method to target workplace inspections. It is recommended that routine inspection be instituted for all companies in which an employee is reported to have a blood lead of 30 microg/dL or greater. PMID- 11434444 TI - Characterization of metalworking fluid aerosols in bearing grinding operations. AB - The concentrations and size distribution of metalworking fluid aerosols were investigated in grinding operations in the bearing manufacturing industry. Fifteen paired open- and closed-face cassette samples and five cascade impactor samples were obtained in each of three types of grinding machinery (face, microcentric, and progressive). Aerosol mass concentration as measured by open face filter sampling ranged from 0.34 to 2.43 mg/m3. As measured by closed-face sampling the range was 0.14 to 2.01 mg/m3. For each grinding process, open-face concentration was significantly higher than the closed-face concentration (paired t-test, p <0.05). Mass median aerodynamic diameter (MMAD) ranged from 3.33 to 6.26 microm. The percentage of mass greater than 9 microm ranged from 8.0 to 45.3. The MMAD and fraction greater than 9 microm were significantly greater for the aerosol produced by the face grinder compared with the other two processes. The results indicate that (1) closed-face sampling results in a lower aerosol mass concentration, as compared with open-face sampling, with the degree of difference being somewhat dependent on grinding process; and (2) the particle size distribution and concentration of metalworking fluid aerosols may vary with the type of grinding operation sampled. PMID- 11434445 TI - PAT Program Report: background and current status. Proficiency Analytical Testing. PMID- 11434446 TI - ELPAT Program Report: background and current status. Environmental Lead Proficiency Analytical Testing. PMID- 11434447 TI - Acute respiratory distress syndrome: pharmacological treatment options in development. AB - The acute respiratory distress syndrome (ARDS) is a clinical syndrome with primarily supportive management options. Despite extensive basic and clinical investigations, multiple pharmacological and nonpharmacological modalities have been unsuccessful in decreasing mortality. Nonetheless, these efforts have substantially heightened our understanding of ARDS pathophysiology. Investigators continue to create new and more complex therapeutic strategies that may have significant clinical impact. Several pharmacological agents for ARDS are in development and have shown either great promise or are at most, under phase II evaluation. The order in which therapeutic options are presented in this review highlights therapeutic options other than the anti-inflammatory approach. In addition to the anti-inflammatory category, vasodilators, surfactant therapy, immunonutrition and partial liquid ventilation are all being evaluated. Within the anti-inflammatory category. new mechanistic approaches include the 'anti inflammatory nature' of interleukin-10, the inhibitory aspects of lysophosphatidic acid on endothelial cell permeability, and the use of recombinant human anti-coagulant proteins (activated protein C and tissue factor pathway inhibitor) to reduce the inflammatory cycle that contributes to microvascular thrombi. Previous work with surfactant in ARDS had its limitations, however, these trials were of sufficient success to spawn 2 new synthetic compounds. These new synthetic surfactants incorporate mixtures of phosphatidylcholine and phosphatidylglycerol (the key phospholipids within endogenous surfactant) and either recombinant surfactant protein C or an analogue of surfactant protein B. Recently, the ARDS Network's low tidal volume study has broken the cycle of decades of negative ARDS trials and demonstrated an improvement in mortality. Through better mechanistic approach and study design, investigator compliance with exclusion criteria, and better understanding of the complexities of patient management, the next pharmacological ARDS trials will hopefully be successful and lead to further reductions in patient mortality. PMID- 11434448 TI - Chemoprevention of breast cancer with fenretinide. AB - Chemoprevention of cancer represents a challenge for oncology during this new millennium. Substantial advances have been accomplished in the last decade, especially for primary and secondary prevention of breast cancer. In addition to tamoxifen, raloxifene and other selective estrogen receptor modulators, retinoids are among the most promising agents, given their ability to inhibit mammary carcinogenesis in preclinical models. Fenretinide, the synthetic amide of retinoic acid, inhibits cell growth mostly through the induction of apoptosis with mechanisms which may partly involve the retinoid receptors. Because it has a favourable toxicological profile, fenretinide has been extensively investigated in clinical trials. A large randomised phase III trial for secondary breast cancer prevention has been recently carried out in Italy. Results showed a reduction of second breast malignancies in premenopausal women. In addition, a significant decrease of circulating insulin-like growth factor (IGF)-1, a known risk factor for premenopausal breast cancer, was observed after 1 year of fenretinide administration in premenopausal women with breast cancer. Ongoing studies on the validation of the circulating IGF-1 as a surrogate endpoint biomarker of fenretinide activity and on the effectiveness of the combination with low dose tamoxifen may provide further insight into the future clinical application of fenretinide. PMID- 11434450 TI - Monotherapy versus combination therapy as first line treatment of uncomplicated arterial hypertension. AB - Mild to moderate hypertension still remains poorly controlled. This relates to multiple factors including low antihypertensive efficacy of single drug therapies reluctance of primary care physicians to modify/titrate initially chosen therapy to obtain target blood pressure, and poor compliance with medication. Several guidelines for the treatment of high blood pressure now include combination therapy with low doses of 2 drugs as one of the strategies for the initial management of mild/moderate arterial hypertension. Evidence discussed in this article points to superior control of blood pressure by combinations of low doses of 2 drugs as compared with monotherapy in regular doses. This superior effectiveness of combined therapy relates to a better antihypertensive efficacy and higher response rates in the low range of doses as the result of complementary mechanisms of antihypertensive effects, better tolerance as a result of a lower rate of adverse effects in the low dose range, improved compliance from better tolerance and simple drug regimen, and lower cost. Whether increased use of fixed low dose combination therapies would translate to better control of arterial hypertension in the population and thereby further reduction of cardiovascular/cerebrovascular morbidity and mortality caused by hypertension remains to be assessed. PMID- 11434449 TI - Newer neuromuscular blocking agents: how do they compare with established agents? AB - Rapacuronium bromide (rapacuronium; ORG-9487) is a nondepolarising muscle relaxant (NMBA) with a low potency [90% effective dose (ED90) 1 mg/kg], which to some extent is responsible for its rapid onset of action. Because of the high plasma clearance (5.3 to 11.1 mg/kg/min) of rapacuronium, its clinical duration of action following single bolus doses up to 2 mg/kg in adults is short (i.e. <20 minutes). Rapacuronium forms a pharmacologically active 3-desacetyl metabolite, ORG-9488, which may contribute to a delay in spontaneous recovery after repeat bolus doses or infusions. After rapacuronium 1.5 mg/kg clinically acceptable intubating conditions are achieved within 60 to 90 seconds in the majority of adult and elderly patients undergoing elective anaesthesia. However, in a rapid sequence setting. intubating conditions are less favourable after rapacuronium 1.5 to 2.5 mg/kg than after succinylcholine. The most prominent adverse effects of rapacuronium (tachycardia, hypotension and bronchospasm) are dose-related, and in particular pulmonary adverse effects are observed more frequently under conditions of a rapid-sequence induction in adults. Therefore, it seems worthwhile to consider only doses of rapacuronium < or = 1.5 mg/kg to facilitate rapid tracheal intubation, and to use succinylcholine or rocuronium rather than rapacuronium in a rapid-sequence setting. Rapacuronium, however, is a suitable alternative to mivacurium chloride (mivacurium) and succinylcholine for short procedures (e.g. ambulatory anaesthesia). Rocuronium bromide (rocuronium) is a relatively low-potent, intermediateacting NMBA. Its main advantage is the rapid onset of neuromuscular block whereby good or excellent intubating conditions are achieved within 60 to 90 seconds after rocuronium 0.6 mg/kg (2 x ED95), and within 60 to 180 seconds after smaller doses (1 to 1.5 x ED95). Larger doses of rocuronium (> or = 1 mg/kg) seem to be suitable for rapid-sequence induction under relatively light anaesthesia. However, it is still a matter of controversy whether, in the case of an unanticipated difficult intubation, the long duration of rocuronium administered in such large doses outweighs the many adverse effects of succinylcholine. Rocuronium has mild vagolytic effects and does not release histamine, even when administered in large doses. Rocuronium is primarily eliminated via the liver and its pharmacokinetic profile is similar to that of vecuronium bromide (vecuronium). Unlike vecuronium, rocuronium has no metabolite. Cisatracurium besilate (cisatracurium), the IR-cis, 1'R-cis isomer of atracurium besilate (atracurium) is approximately 4 times more potent than atracurium. The onset time of cisatracurium is significantly slower than after equipotent doses of atracurium. The recommended intubating dose is 0.15 to 0.2 mg/kg (3 to 4 times ED95). Over a wide range of clinically relevant doses the recovery properties of cisatracurium are affected by neither the size of the bolus dose nor by the duration of infusion. Unlike atracurium, cisatracurium does not trigger histamine release. Like atracurium, cisatracurium undergoes Hofmann elimination. In contrast to atracurium, cisatracurium does not undergo hydrolysis by nonspecific plasma esterases. Moreover, about 77% of the drug is cleared by organ-dependent mechanisms. PMID- 11434452 TI - Current management of ethylene glycol poisoning. AB - Ethylene glycol, a common antifreeze, coolant and industrial solvent, is responsible for many instances of accidental and intentional poisoning annually. Following ingestion, ethylene glycol is first hepatically metabolised to glycoaldehyde by alcohol dehydrogenase. Glycoaldehyde is then oxidised to glycolic acid, glyoxylic acid and finally oxalic acid. While ethylene glycol itself causes intoxication, the accumulation of toxic metabolites is responsible for the potentially fatal acidosis and renal failure, which characterises ethylene glycol poisoning. Treatment of ethylene glycol poisoning consists of emergent stabilisation, correction of metabolic acidosis, inhibition of further metabolism and enhancing elimination of both unmetabolised parent compound and its metabolites. The prevention of ethylene glycol metabolism is accomplished by the use of antidotes that inhibit alcohol dehydrogenase. Historically, this has been done with intoxicating doses of ethanol. At a sufficiently high concentration, ethanol saturates alcohol dehydrogenase, preventing it from acting on ethylene glycol, thus allowing the latter to be excreted unchanged by the kidneys. However, ethanol therapy is complicated by its own inherent toxicity, and the need to carefully monitor serum ethanol concentrations and adjust the rate of administration. A recent alternative to ethanol therapy is fomepizole, or 4-methylpyrazole. Like ethanol, fomepizole inhibits alcohol dehydrogenase; however it does so without producing serious adverse effects. Unlike ethanol, fomepizole is metabolised in a predictable manner, allowing for the use of a standard, validated administration regimen. Fomepizole therapy eliminates the need for the haemodialysis that is required in selected patients who are non acidotic and have adequate renal function. PMID- 11434453 TI - Barnidipine. AB - Bamidipine is an antihypertensive drug belonging to the dihydropyridine (DHP) group of calcium antagonists. It is available in a modified-release formulation which has a gradual onset of action and is effective in a single daily oral dose of 10 to 20 mg. Bamidipine has selective action against cardiovascular calcium antagonist receptors and its antihypertensive action is related to the reduction of peripheral vascular resistance secondary to its vasodilatory action. The clinical antihypertensive efficacy of barnidipine is similar to that of other DHP calcium antagonists such as nitrendipine and amlodipine, and antihypertensives belonging to other drug classes such as atenolol and enalapril. Barnidipine has been found to be as efficacious and well tolerated as hydrochlorothiazide in the management of hypertension in elderly patients. Barnidipine is generally well tolerated. As with other DHP calcium antagonists, vasodilator adverse events such as headache, flushing and peripheral oedema account for most of the adverse events reported with its use and are usually transient. Oedema is less frequent than with amlodipine and nitrendipine. Its use is not associated with reflex tachycardia. PMID- 11434451 TI - Strategies for the treatment of cancer pain in the new millennium. AB - As was the case in the era before us, in the new millennium we will continue to see an abundance of patients experiencing cancer-related pain for different reasons. Although much needless pain and suffering still affects many of those with cancer, we are presented with a medical dichotomy. With the analgesic drugs available today, and the relatively simple and effective guidelines to treat cancer pain published and disseminated by the World Health Organization, why do people with cancer continue to experience pain? As we search for the answer, the horizon may hold promising new drugs, 'old drugs' with new interest and applications, and new strategies for the field of pain therapy. Possibilities include the isolation and development of analgesics or analgesic combinations that may minimise the adverse effects which are often associated with the current therapeutic class of opioid analgesics. In addition, current research points to promising results identifying the N-methyl D-aspartate non-opioid receptor as a likely component of neuropathic pain. Drugs such as gabapentin, the mechanism of action of which is not well known, have found favour within the clinical community for their analgesic properties and good tolerability. Methadone, in a phase of resurgence, has garnered the attention of the clinical community because of its unique receptor activity and pharmacoeconomic benefits. A number of clinical studies have demonstrated that methadone has a valuable role in treating cancer pain. Perhaps, an unbalanced focus on the risks of inappropriate use, rather than the benefits, should not compromise or distract from the use of methadone as an alternative to morphine. Studies are on going to assess the potential role of methadone in treating neuropathic pain. Drugs such as cannabinoids, although currently applicable for patients with anorexia, nausea and/or vomiting, may offer benefits to patients experiencing pain. Other opportunities exist with such compounds as alpha2-adrenergic agonists, nicotine, lidocaine and ketamine. New strategies such as the switching opioids and/or their route of administration may offer improved analgesia with fewer adverse effects, thus providing therapeutic alternatives for the clinical community. In addition, there is interest in the co-administration of opioids that act on different receptors. For instance, oxycodone appears to be a kappa opioid receptor agonist and may offer enhanced analgesia when combined with morphine. PMID- 11434455 TI - Barley chromosome addition lines of wheat for screening of AFLP markers on barley chromosomes. AB - We conducted AFLP (Amplified Fragment Length Polymorphism) analysis with the six wheat-barley chromosome addition lines of common wheat cultivar Chinese Spring. We analyzed the AFLP fingerprints generated by 36 combinations of selective amplification primers to find 103 markers specific to the barley chromosomes (2.9 markers per combination on average). The numbers of AFLP markers mapped to the barley chromosomes varied (one to 16) depending of the primer combinations. Each barley chromosome had 10 to 27 AFLP markers (17.2 markers on average). We identified the chromosome arms in which these markers are located using the barley telocentric addition lines (one to 20 markers per chromosome arm). The AFLP markers were not distributed evenly among chromosomes and chromosome arms. We could not determine the chromosome-arm locations for some of the barley specific markers, either because such markers were found in both the short- and long-arm telocentric lines, or in neither line. PMID- 11434456 TI - Characterization of cryptic flagellin genes in Shigella boydii and Shigella dysenteriae. AB - Flagellin (fliC) genes of 12 Shigella boydii and five Shigella dysenteriae strains were characterized. Though these strains are nonmotile, the cryptic fliCSB gene, cloned from S. boydii strain C3, is functional for expression of flagellin. It consists of 1,704 bp, and encodes 568 amino acid residues (57,918 Da). The fliCSD gene from S. dysenteriae strain 16 consists of 1,650 bp encoding 549 amino acid residues (57,591 Da) and contains an IS1 element inserted in its 3' end. The two genes are composed of the 5'-constant, central variable and 3' constant sequences, like other known fliC genes. The two genes share high homology in nucleotide and amino acid sequences with each other and also with the Escherichia coli fliCE gene, indicating that both genes are closely related to the fliCE gene. Comparison of the central variable sequences of six different fliC genes showed that the fliCSB and fliCSD genes share low homology in amino acid sequence with the other fliC genes, suggesting that they encode antigenic determinants intrinsic to respective subgroups. However, Southern blotting using as probes the central variable sequences of several fliC genes showed that four of 12 S. boydii strains have a fliC gene similar to that of Shigella flexneri, and that among five fliC genes from S. dysenteriae strains, one is similar to that of S. flexneri, two are similar to that of S. boydii, and only one is unique to S. dysenteriae. Some of these variant alleles were verified by immunoblotting with flagellins produced from cloned fliC genes. The presence of variant fliC alleles in S. boydii and S. dysenteriae indicates that subdivision into subgroups does not reflect the ancestral flagella H antigenic relationships. These data will be useful in considering the evolutionary divergence of the Shigella spp.. PMID- 11434454 TI - Alendronate: an update of its use in osteoporosis. AB - Alendronate (alendronic acid) is a nitrogen-containing bisphosphonate which binds to bone surfaces and inhibits bone resorption by osteoclasts. Oral alendronate 5 or 10 mg/day produces sustained increases in bone mineral density (BMD) in postmenopausal women with or without osteoporosis, in men with primary osteoporosis and in both men and women with or without osteoporosis receiving systemic corticosteroid therapy. Histomorphometric analyses have found that alendronate does not appear to impair bone quality. Alendronate reduced the risk of radiographic vertebral fracture, clinical vertebral fracture or hip fracture by 47 to 56% in postmenopausal women who had > or = 1 existing vertebral fracture and in those with no existing vertebral fractures but who had osteoporosis. In a number of comparative trials in postmenopausal women with osteoporosis, alendronate 10 mg/day was found to be more effective at inducing sustained increases in BMD than intranasal calcitonin, and at least as effective as conjugated estrogens and raloxifene. Alendronate 70 mg administered once weekly and 35 mg twice weekly are as effective at increasing BMD as 10 mg/day in this patient group. In clinical trials, alendronate was generally well tolerated when taken as recommended. Adverse events tended to be transient and associated with the upper GI tract, most commonly including abdominal pain, nausea, dyspepsia, acid regurgitation and musculoskeletal pain. No statistically significant differences between alendronate 10 mg/day and placebo have been found in the incidence of upper GI adverse events in large clinical trials. However, postmarketing surveillance reported a low incidence of adverse events related to the oesophagus. Specific instructions aimed at reducing the risk of upper GI adverse events have been provided by the manufacturer. CONCLUSIONS: Alendronate is effective and generally well tolerated in the treatment of women or men with primary (including postmenopausal) or corticosteroid-induced osteoporosis and in the prevention of osteoporosis in postmenopausal women. The drug has been associated with upper GI tract adverse events, although the extent to which alendronate is responsible for these events has not been clearly established. Alendronate should be considered a treatment of choice in postmenopausal women with osteoporosis. Alendronate is also a suitable treatment option for primary osteoporosis in men and for corticosteroid-induced osteoporosis in both men and women. PMID- 11434457 TI - Sequence characterization of the vir region of a nopaline type Ti plasmid, pTi SAKURA. AB - We isolated a crown gall tumor-inducing nopaline type Ti plasmid from Agrobacterium tumefaciens on a Sakura Japanese cherry tree, and designated it as pTi-SAKURA. By primer walking sequencing with long PCR and a newly developed PCR subcloning technique for long insert DNA, we completed DNA sequencing of the most important functional unit, the virulence (vir) region of pTi-SAKURA, which is indispensable for T-DNA transfer into the plant's chromosomes. By homology searches with the vir genes of other bacterial plasmids, we identified 11 open reading frames (orfs) and 31 genes and 11 vir box, which are 6 bp regulatory sequences. In total, 26 vir genes, including the putative virF and virK and the main vir region, were present as the vir gene cluster. The presence of vir box, GC content, codon usage and expression analysis in these genes led us to propose a new vir region. PMID- 11434458 TI - The transposition pattern of the Ac element in tobacco cultured cells. AB - We investigated physical distances and directions of transposition of the maize transposable element Ac in tobacco cultured cells. We introduced a T-DNA construct that carried a non-autonomous derivative of Ac (designated dAc-I-RS) that included sites for cleavage by restriction endonuclease MluI. Another cleavage site was also introduced into the T-DNA region outside of the dAc-I-RS transposable element. The tobacco cultured cell line BY-2 was transformed with the T-DNA and several transformed lines that had a single copy of the T-DNA at a different chromosomal location were isolated. These lines were co-cultured with Agrobacterium tumefaciens cells that carried a cDNA for the Ac transposase gene under the control of various promoters. Sublines of cultured cells in which dAc-I RS had been transposed, were isolated. The genomic DNAs of these sublines were isolated and digested with MluI. Sizes of DNA segments generated by digestion were determined by pulse-field gel electrophoresis. Our results showed that 20 to 70% of transposition events had occurred within several hundreds kilo-base pairs (kb) on the same chromosome. These results demonstrate that the Ac-Ds element preferentially transposed to regions near the original site in a tobacco chromosome. In addition, the present results are an example of asymmetric transposition as demonstrated by the distance of transposition on the chromosome. PMID- 11434459 TI - Regulation of the localization of Dbf2 and mob1 during cell division of saccharomyces cerevisiae. AB - The mitotic exit network (MEN) governs Cdk inactivation. In budding yeast, MEN consists of the protein phosphatase Cdc14, the ras-like GTPase Tem1, protein kinases Cdc15, Cdc5, Dbf2 and Dbf2-binding protein Mob1. Tem1, Dbf2, Cdc5 and Cdc15 have been reported to be localized at the spindle pole body (SPB). Here we report changes of the localization of Dbf2 and Mob1 during cell division. Dbf2 and Mob1 localize to the SPBs in anaphase and then moves to the bud neck, just prior to actin ring assembly, consistent with their role in cytokinesis. The neck localization, but not SPB localization, of Dbf2 was inhibited by the Bub2 spindle checkpoint. Cdc14 is the downstream target of Dbf2 in Cdk inactivation, but we found that the neck localization of DbP2 and Mob1 was dependent on the Cdc14 activity, suggesting that Dbf2 and Mob1 function in cytokinesis at the end of the mitotic signaling cascade. PMID- 11434460 TI - Genomic organization, transcription start sites, and chromosomal location of the Drosophila cortactin gene. AB - An actin filament binding protein cortactin was initially identified as a major phosphotyrosine-containing protein in v-Src-transformed chicken embryo fibroblast cells. The mouse, human, and Drosophila homologs were independently identified as a signaling molecule involved in a mitogenic response, as a product of a putative oncogene EMS1, and as a molecule interacting with a scaffolding protein ZO-1, respectively. In this report, we describe the cloning of the Drosophila cortactin gene, which consists of four exons and three introns, covering 3 kilobases in length. All exon-intron junctions are well matched with the GT/AG consensus sequence. S1 nuclease mapping revealed one major and several minor transcription start sites. The cytological location of the Drosophila cortactin gene is between chromosome segments 93B3 and 93B7. PMID- 11434461 TI - RAPD, RFLP and SSLP analyses of phylogenetic relationships between cultivated and wild species of rice. AB - RAPD, RFLP, nuclear SSLP and chloroplast SSLP analyses were carried out to clarify the phylogenetic relationships among A-genome species of rice. In total, 12 cultivars of Oryza sativa (4 Japonica, 3 Javanica and 5 Indica), one cultivar of O. glaberrima, and 17 wild accessions (12 O. rufipogon, 2 O. glumaepatula, 1 O. longistaminata, 1 O. meridionalis and 1 O. barthii) were used. Their banding patterns were scored and compared to evaluate the similarity between accessions. Genetic differentiation within and between taxa was examined based on the average similarity indices. Except for chloroplast SSLP analysis, the average similarities were higher within O. sativa than within O. rufipogon, and O. sativa Indica had greater intrasubspecific variation than Japonica and Javanica. Comparisons between cultivated and wild species showed that O. sativa was closely related to O. rufipogon, while O. glaberrima was closely related to O. barthii. This indicated that two cultivated species, O. sativa and O. glaberrima, originated from O. rufipogon and O. barthii, respectively. Domestication of O. sativa seemed to be diphyletic, since strong similarity was observed between O. sativa Japonica-Javanica and O. rufipogon from China and between O. sativa Indica and O. rufipogon from tropical Asia. In addition, dendrograms for RAPD, RFLP, and nuclear and chloroplast SSLP analyses were constructed to reveal the overall genetic relationships among A-genome species. In all analyses, O. sativa and O. glaberrima formed groups with O. rufipogon and O. barthii, respectively. However, their manners of clustering with other wild species were not the same. The results of RAPD and RFLP analyses indicate that O. glumaepatula was relatively close to the groups of O. sativa and O. glaberrima whereas O. longistaminata and O. meridionalis were highly differentiated from other A-genome species. On the other hand, clear interspecific relationships were not obtained by nuclear or chloroplast SSLP analyses. PMID- 11434462 TI - A mixed model method to predict QTL-cluster effects using trait and marker information in a multi-group population. AB - In this study, a mixed model method using trait phenotype and marker information was developed for genetic evaluation of animals in a crossbred population originated from several founder genetic groups. The situation in which a cluster of QTLs is located in a particular chromosome region and is marked by two flanking markers is considered. With this method, the conditional expectation of the identity-by-descent proportion for the QTL-cluster marked and the genetic variances and covariances, given genetic group and marker information, are properly taken into account. The structure of segregation variance used in this method is different from that in the case of a single QTL marked. The current method provides best linear unbiased estimation of the relevant fixed effects and best linear unbiased prediction of the additive effects for the QTL-cluster marked and of the additive effects of the remaining polygenes. A small numerical example is given to illustrate the current prediction procedure. PMID- 11434463 TI - The gene for alternative oxidase-2 (AOX2) from Arabidopsis thaliana consists of five exons unlike other AOX genes and is transcribed at an early stage during germination. AB - We investigated the expressions of genes for alternative oxidase (AOX1a, AOX1b, AOX1c and AOX2) and genes for cytochrome c oxidase (COX5b and COX6b) during germination of Arabidopsis thaliana, and examined oxygen uptakes of the alternative respiration and the cytochrome respiration in imbibed Arabidopsis seeds. A Northern blot analysis showed that AOX2 mRNA has already accumulated in dry seeds and subsequently decreased, whereas accumulation ofAOX1a mRNA was less abundant from 0 hours to 48 hours after imbibition and then increased. The increase of the capacity of the alternative pathway appeared to be dependent on the expressions of both AOX2 and AOX1a. On the other hand, steady-state mRNA levels of COX5b and COX6b were gradually increased during germination, and the capacity of the cytochrome pathway was correlated with the increase of expressions of the COX genes. Antimycin A, the respiratory inhibitor, strongly increased the expression of AOX1a but had no effect on the expression of AOX2. A 5'RACE analysis showed that AOX2 consists of five exons, which is different from the case of most AOX genes identified so far. Analysis of subcellular localization of AOX2 using green fluorescent protein indicated that the AOX2 protein is imported into the mitochondria. PMID- 11434465 TI - Rheumatology in sub-Saharan Africa. PMID- 11434464 TI - Structure and characterization of the gene encoding alpha subunit of soybean beta conglycinin. AB - beta-conglycinin, a soybean seed storage protein, is comprised of three different subunits, a, alpha', and beta. Several candidates for the alpha subunit gene have been isolated, however, the structure of the alpha subunit gene has not been completely determined. Accordingly, it was also unknown which of the gene candidates are functionally active. Here, we have determined the nucleotide sequence and transcription start site of the alpha subunit gene, and compared the structural components with those of the other subunits or other seed protein genes. The a subunit gene, which is located on a 7.6-kb EcoRI fragment, was composed of six exons that had the same organization as those for the alpha' subunit gene. Within a 400 bp upstream region of the transcription start site, four regions (designated as boxes I, II, III, and IV) were found to be conserved among the alpha, alpha', and other seed protein genes. Genomic Southern blot analysis of soybean varieties lacking the alpha subunit gene candidate indicated that the gene characterized in this paper actually encodes the a subunit and is functionally active. In addition, these experiments revealed the presence of an additional gene which is also responsible for the expression of the a subunit. PMID- 11434466 TI - Pulmonary function tests, aerobic capacity, respiratory muscle strength and endurance of patients with rheumatoid arthritis. AB - Patients with rheumatoid arthritis (RA) show lower cardiorespiratory fitness than normal subjects. This study was planned to investigate the pulmonary function tests (PFT), respiratory muscle strength and endurance, and aerobic capacity of patients with RA, as well as the relationship of these parameters to clinical and functional status. Twenty-five RA patients aged 25-71 (48.52 +/- 14.09) and 21 control subjects aged 25-66 (45.67 +/- 13.27) participated in the study. PFT, maximum volunteer ventilation, maximum inspiratory and maximum expiratory pressures and cardiorespiratory exercise tests were carried out in all subjects to evaluate the respiratory involvement, inspiratory and expiratory muscle strength and endurance, and aerobic capacity. Patients' duration of disease, smoking and alcohol habits, duration of morning stiffness, visual analogue scale scores, ARA functional classifications and Ritchie articular indexes were recorded. All the patients and control subjects were non-exercising individuals. As a result, we found that RA patients have normal PFT but reduced respiratory muscle strength and endurance, and also reduced aerobic capacity compared to controls. According to this result, respiratory and aerobic exercises may be recommended to improve respiratory muscle strength and endurance and aerobic capacity in these patients. PMID- 11434467 TI - Capillaroscopic findings in erosive and nodal osteoarthritis of the hands. AB - Osteoarthritis of the hands is a very common disease that can present a large number of different clinic pictures, such as nodal (NOA) and erosive (EOA) forms. EOA in particular is a rare subset of hand osteoarthritis characterised by faster destructive changes involving the distal interphalangeal (DIP) and proximal interphalangeal (PIP) joints. In the early stages of the disease the differential diagnosis from other arthritides, such as rheumatoid or seronegative arthritis, may pose a challenge. Nailfold capillaroscopy is a non-invasive technique that allows the in vivo study of the microvascular environment. In this study the authors have compared the capillaroscopic microvascular patterns in 56 patients with EOA, in 46 patients with NOA, and in 50 normal controls. The abnormalities that could be found in EOA patients were similar to those described by some authors in patients with psoriatic arthritis. The authors discuss the significance of these abnormalities and the possible relationship between EOA and psoriatic arthritis. PMID- 11434468 TI - Static lung compliance as an index of early pulmonary disease in systemic sclerosis. AB - Pulmonary function abnormalities constitute the most important cause of morbidity and mortality in patients with systemic sclerosis (SSc). A restrictive ventilatory defect with a reduction in pulmonary carbon monoxide diffusion capacity (DLCO) is the commonest finding. Static lung compliance (Cst) is perhaps the most sensitive parameter for the detection of abnormal pulmonary mechanics. We compared the results of Cst and DLCO in patients with SSc and normal spirometry to identify early pulmonary disease. Sixteen of 50 patients had normal spirometry, eight of whom had abnormal Cst. Four of them also had a reduced DLCO. Two patients had abnormal DLCO but their Cst was normal. One of them had pulmonary hypertension on echocardiography. We conclude that Cst was helpful in diagnosing early pulmonary abnormality in 50% of patients with normal spirometry. Among these patients Cst was a better index of abnormality than DLCO. PMID- 11434469 TI - The clinical efficacy of low-power laser therapy on pain and function in cervical osteoarthritis. AB - Pain is a major symptom in cervical osteoarthritis (COA). Low-power laser (LPL) therapy has been claimed to reduce pain in musculoskeletal pathologies, but there have been concerns about this point. The aim of this study was to evaluate the analgesic efficacy of LPL therapy and related functional changes in COA. Sixty patients between 20 and 65 years of age with clinically and radiologically diagnosed COA were included in the study. They were randomised into two equal groups according to the therapies applied, either with LPL or placebo laser. Patients in each group were investigated blindly in terms of pain and pain related physical findings, such as increased paravertebral muscle spasm, loss of lordosis and range of neck motion restriction before and after therapy. Functional improvements were also evaluated. Pain, paravertebral muscle spasm, lordosis angle, the range of neck motion and function were observed to improve significantly in the LPL group, but no improvement was found in the placebo group. LPL seems to be successful in relieving pain and improving function in osteoarthritic diseases. PMID- 11434470 TI - No evidence of cardiac autonomic involvement in ankylosing spondylitis, as assessed by heart rate variability. AB - The aim of this study was to investigate the involvement of autonomic nervous system (ANS) function by using power spectral analysis of heart rate variability (HRV) method in patients with ankylosing spondylitis (AS). The study included 94 AS patients all fulfilling the New York criteria for AS, and 49 healthy volunteers. Recordings for HRV were obtained with a PC-based high-resolution electrocardiographic system and analysed using power spectral analysis. The peak around 0.04-0.15 Hz was defined as low-frequency peak (LF) and the other, around 0. 15-0.40 Hz, was defined as high-frequency peak (HF), representing mostly the sympathetic and the parasympathetic components of the ANS, respectively. The following variables were calculated and compared between groups: the LF in absolute and normalised units (LF nU); the HF in absolute and normalised units (HF nU); and LF/HF ratio. The AS group included 47 male and 47 female subjects with a mean age of 33 +/- 11 years (range 16-64). In the control group there were 23 male and 26 female healthy subjects (mean age 33 +/- 8; range 19-60). None of the patients or control subjects had any cardiac or neurological symptoms. Both groups were similar with respect to age and sex characteristics (p > 0.05). The HRV analysis indicated that the peaks of LF, LF nU, HF, HF nU and LF/HF ratio were similar in both groups. Groups also did not differ with respect to heart rate at the time of examination. Our data demonstrated no evidence of ANS involvement as assessed by HRV analysis in AS patients. PMID- 11434471 TI - A controlled study of sacroiliitis in Behcet's disease. AB - The aim of the study was to evaluate the prevalence of sacroiliitis in a group of patients with Behcet's disease (BD). Pelvic X-rays of 27 patients with BD responding to the International Study Group of BD and 30 controls (15 AS and 15 sciatica) were read blind and sacroiliac involvement was graded according to the New York criteria. In a second step, patients or controls with equivocal sacroiliitis had a sacroiliac CT scan. Two patients with BD (7.4%) and all patients with AS had evident bilateral sacroiliitis (at least grade 2). One patient with BD and two patients with sciatica had equivocal sacroiliitis (grade 1). CT confirmed sacroiliitis in the two patients with BD and eliminated inflammatory sacroiliitis in the three other patients with equivocal sacroiliitis showing mild degenerative lesions. A review of the literature showed that sacroiliitis and AS are rarely associated with BD. There remains insufficient evidence to suggest that sacroiliitis is an intrinsic feature of BD and that BD belongs to the group of SpA. PMID- 11434472 TI - Why do patients with rheumatoid arthritis use alternative treatments? AB - The aim of this study was to analyse the characteristics of patients with rheumatoid arthritis (RA) who make use of alternative or complementary medicine (CM). Two hundred and sixty-two randomly chosen patients with RA filled out self assessment health status and pain questionnaires. Differences between the group of patients making use of both CM and conventional treatment (n = 52) and the group of patients who relied only on conventional treatment prescribed by their rheumatologists (n = 210) were explored with respect to demographic characteristics, duration of RA, levels of physical, psychological and social functioning, and pain-coping behaviour. We found that female patients used CM more often than did male patients, and those who used CM were younger than those who did not. There were no differences with respect to duration of RA, physical, psychological or social functioning or pain coping; however, the perceived impact of RA on several domains of life was higher in patients who used CM than in those who did not. Nevertheless, the patient groups did not differ in terms of medical consumption, except that those who used CM visited medical specialists for RA related complaints less than those who relied only on conventional treatments. We concluded that the higher impact of RA, in the absence of worse disease, perceived by users of CM in several domains of life, especially psychosocial functioning, could be the reason they use CM. This suggests that CM cannot be substituted by additional conventional treatment prescribed by the rheumatologist, but rather by psychosocial intervention. PMID- 11434473 TI - Adult-Onset Still's disease in Saudi Arabia. AB - We present a series of 14 Saudi patients diagnosed to have adult-onset Still's disease (AOSD). The clinical and laboratory pattern of AOSD in our series is more or less similar to that in other reported series, apart from having lower cardiac and pulmonary involvement than western series. The disease course was relatively benign, with only half the cases showing recurrences, which were controlled by alterations in the dosage of corticosteroid and NSAIDs. A comparison with other series is given. PMID- 11434474 TI - Undifferentiated spondyloarthropathies: a 2-year follow-up study. AB - The aim of the study was to analyse the 2-year follow-up of a series of patients with the diagnosis of undifferentiated spondyloarthropathy (uSpA). A prospective study was carried out analysing 68 patients with symptomatic uSpA who fulfilled the European Spondylarthropathy Study Group (ESSG) criteria for seronegative spondyloarthropathies (SpA) and were aged between 18 and 50 years. Inclusion criteria included inflammatory low back pain (ILBP) (without radiographic sacroiliitis), asymmetric oligoarthritis (predominantly affecting large joints in the lower limbs) and heel enthesopathies (Achilles tendinitis and/or plantar fasciitis). Imaging methods included pelvic radiography (at study entry and after 2 years) and calcaneal radiography (at study entry). There was a predominance of male gender (78%), caucasoid race (72%) and positive HLA-B27 (54%), with a mean age of 31 years and mean disease duration of 5 years. The first disease manifestations were ILBP (49%), asymmetric oligoarthritis (35%) and heel enthesopathies (16%). A positive family history of a definite SpA was mentioned by 9% of the patients. Seventeen patients (25%) scored 5 points in the Amor set of SpA criteria; logistic regression analysis showed that HLA-B27, heel enthesopathy and asymmetric oligoarthritis were significantly associated with Amor criteria > or = 6, whereas ILBP was associated with Amor criteria <6. Male sex was associated with heel enthesopathies (p = 0.041) and ankle involvement (p = 0.015). Caucasoid race was associated with ILBP (p=0.015) and buttock pain (p = 0.047). Positive HLA-B27 was associated with wrist involvement (p=0.019) and Amor criteria > or = 6 (p=0.001). After a 2-year follow-up the following outcomes were observed: uSpA 75%; disease remission 13%; ankylosing spondylitis 10%; psoriatic arthritis 2%. Logistic regression analysis showed that buttock pain and positive HLA-B27 (trend) were statistically associated with progression to a definite SpA. In conclusion, uSpA can represent a provisional diagnosis in the group of SpA and a systematic follow-up is necessary in order to better establish the different patterns of the disease. PMID- 11434475 TI - Associations of isokinetic knee extensor and flexor strength with steroid use and walking ability in women with rheumatoid arthritis. AB - Seventy-five women with rheumatoid arthritis according to the 1987 criteria of the American Rheumatism Association were examined. Mean age was 61.9 +/- 12.5 years, mean disease duration 14 years. Sixty-three were or had been on steroids (median cumulative prednisolone dose 2.5 g). Maximal voluntary knee extensor and flexor strength (Nm) was assessed at 30 degrees/s by an isokinetic dynamometer. Walking ability was expressed as walking and stair-climbing time (s). Markers of disease activity included number of swollen and tender joints, pain as recorded by the patients on a visual analogue scale (VAS), and disability as scored by the Stanford Health Assessment Questionnaire (HAQ). Muscle strength, walking time (50 m) and stair-climbing time were reduced on average by 30%, and increased by 28% and 54% (p<0.0001), respectively, compared to 67 age-, weight- and height-matched healthy women. Associations between muscle strength and cumulative or current steroid dose were not found after correction for age and disease duration. Significant linear correlations were found between knee extensor strength and walking time (r =-0.78, p<0.0001) and stair-climbing time (r = -0.76, p<0.0001). Similar correlations were found for flexor strength. The correlations remained significant (Rpartial ranging from -0.64 to -0.69, p<0.0001) in multiple regression analyses adjusting for age, height, weight, disease duration, number of swollen and tender joints, and VAS and HAQ scores. In conclusion, negative effects of steroids on muscle strength were not demonstrated. Leg muscle strength is an important and independent determinator of walking ability in RA. PMID- 11434476 TI - Ultrasonography and colour doppler sonography of salivary glands in primary Sjogren's syndrome. AB - To examine either the ultrasonographic (US) features of the parotids and submandibular glands or the blood flow alterations that may occur in the salivary glands of patients with primary Sjogren's syndrome (pSS) we studied 30 female patients with pSS and 30 controls suffering from dry mouth not due to pSS. All measurements were taken by the same examiner, who used the same equipment to avoid interobserver variability. The US parameters recorded (parenchymal homogeneity, echogenicity, size of the glands and posterior glandular border) were scored according to a previously described scoring system. For each waveform, peak systolic velocity (PSV) and resistive index (RI) were measured at the external carotid artery in the examination of the parotids and at the facial artery within the submandibular glands before and during lemon juice stimulation. On the basis of the degree of chronic inflammatory changes at minor salivary gland (MSG) biopsy, chronic sialadenitis (CS) was defined as mild in 10 and severe in 20 pSS patients. Abnormal US scores were obtained in 26/30 (86.6%) pSS patients and in 9/30 (30%) controls. Moreover, in pSS patients the US scores were sigificantly higher than in the control group (p=-0.0001). The mean (+/- SD) difference between the PSV values taken from parotids and submandibular glands before and during lemon juice stimulation was statistically significant (p=0.003 and p=0.01, respectively) in the controls. On the other hand, no significant changes in the PSV values were found in the whole group of pSS patients. However, the changes in PSV values before and during lemon juice stimulation were statistically significant in both parotids (p=0.019) and submandibular glands (p=0.012), and not significantly different from those in the controls in pSS patients with mild CS. The variability of RI taken from the salivary glands before and during lemon juice stimulation was not statistically significant in either pSS patients or controls. US abnormalities were detected in the majority of pSS patients and their severity was significantly greater than those recorded in the controls. Of the colour Doppler sonographic (CDS) parameters only PSV was influenced by the degree of chronic inflammation, as shown at the MSG biopsy, suggesting that PSV may reflect the vascular changes occurring in the salivary glands during the course of an autoimmune disease such as pSS. PMID- 11434477 TI - Sudden sensorineural hearing loss as a first manifestation of systemic lupus erythematosus: association with anticardiolipin antibodies. AB - Sudden sensorineural hearing loss is a rarely reported manifestation of systemic lupus erythematosus (SLE). This condition has been most frequently seen in individuals with concomitant anticardiolipin antibody (ACL) syndrome, although a direct causal relationship remains unconfirmed. We report an unusual case of a young male with sudden unilateral hearing loss as the first manifestation of SLE. This individual was also found to be ACL positive and subsequently presented with other thrombotic manifestations compatible with this syndrome. The literature regarding this condition is reviewed and the significance of this case in fortifying the association of anticardiolipin antibodies and sensorineural hearing loss is discussed. PMID- 11434478 TI - Association of Eosinophilic fasciitis and epileptic seizure. AB - Visceral involvement in eosinophilic fasciitis (EF) is a well known but rare event. With regard to neurological manifestations, both carpal tunnel syndrome and peripheral neuropathy have been described. We report the case of a 40-year old woman with EF who had a major motor seizure. This association might not be fortuitous and be related to the known neurotoxin of eosinophils. PMID- 11434479 TI - Remission of the renal involvement in a patient with primary Sjogren's syndrome (SS) after pulse high-dose corticosteroid infusion therapy. AB - We report the case of a young female patient with primary Sjogren's syndrome (SS). In addition to sicca symptoms she also suffered from progressive renal insufficiency and renal tubular acidosis (RTA). She was treated with three sets of pulse high-dose corticosteroid infusion and subsequent low-dose corticosteroid oral administration. When the efficacy was evaluated about 6 months after the start of the therapy, dramatic improvements were seen with no adverse effects, not only in laboratory tests but also histopathologically, as indicated by the repeat kidney biopsy. This suggests that renal involvements of SS might be reversible in some cases, and that there might be a clinical benefit of pulse high-dose corticosteroid infusion therapy in SS with progressive renal involvement. PMID- 11434480 TI - Septic arthritis of the hip due to Fusobacterium nucleatum. AB - Anaerobic bacteria are uncommon pathogens in septic arthritis. We report a case of pyarthrosis of the right hip caused by Fusobacterium nucleatum, following a transient synovitis in an otherwise healthy boy. There are only a few cases involving this species described in the literature. This report illustrates the difficulty of isolating this unusual organism and emphasises the usefulness of the Bactec blood culture bottles for the recovery of anaerobic bacteria. PMID- 11434481 TI - Magnetic resonance imaging findings of manubriosternal joint involvement in SAPHO syndrome. PMID- 11434482 TI - Introduction: new format for executive summary of practice parameters. PMID- 11434483 TI - Practice parameter for the assessment and treatment of children and adolescents with suicidal behavior. American Academy of Child and Adolescent Psychiatry. AB - These guidelines review what is known about the epidemiology, causes, management, and prevention of suicide and attempted suicide in young people. Detailed guidelines are provided concerning the assessment and emergency management of the children and adolescents who present with suicidal behavior. The guidelines also present suggestions on how the clinician may interface with the community. Crisis hotlines, method restriction, educational programs, and screening/ case-finding suicide prevention strategies are examined, and the clinician is advised on media counseling. Intervention in the community after a suicide, minimization of suicide contagion or imitation, and the training of primary care physicians and other gatekeepers to recognize and refer the potentially suicidal child and adolescent are discussed. PMID- 11434484 TI - Practice parameter for the assessment and treatment of children and adolescents with schizophrenia. American Academy of Child and Adolescent Psychiatry. AB - This practice parameter reviews the literature on the assessment and treatment of children and adolescents with schizophrenia. Recommendations are based on the limited research available, the adult literature, and clinical experience. Early onset schizophrenia is diagnosed using the same criteria as in adults, and it appears to be continuous with the adult form of the disorder. Noted characteristics of youth with schizophrenia include predominance in males, high rates of premorbid abnormalities, and often poor outcome. Differential diagnosis includes psychotic mood disorders, developmental disorders, organic conditions, and nonpsychotic emotional/behavioral disorders. Treatment strategies incorporate antipsychotic medications with psychoeducational, psychotherapeutic, and social and educational support programs. The advent of atypical antipsychotic agents has enhanced the potential for effective treatment. PMID- 11434485 TI - Orthopedic problems in sport climbing. AB - Sport climbing is associated with unique upper- and lower-limb injuries involving predominantly the hand, elbow, and shoulder, and to a lesser extent the foot. Many pathologic conditions are limited to sport climbing. Physicians treating sport climbers should be aware of these unique injuries and overuse syndromes. This article presents an overview of orthopedic problems (injuries, overuse syndromes, and fractures) resulting from sport climbing. Sport climbing is defined in the context of existing mountain sports, and its characteristics and technical terms are presented. The etiology, diagnosis, and specific treatment for orthopedic problems associated with sport climbing are described. PMID- 11434486 TI - Use of stun guns for venomous bites and stings: a review. AB - During the past 2 decades, articles suggesting that stun guns be utilized to treat venomous bites and stings have appeared in both the lay and medical press. Although never widely considered to be standard therapy for venomous bites and stings, stun guns are still considered to be a treatment option by some medical practitioners and outdoor enthusiasts. A Medline search was performed using these terms: venomous bites, venomous stings, snake bites, spider bites, electrical, stun gun, high voltage electricity, low amperage electricity, direct current, and shock therapy. Articles selected included laboratory-based isolated venom studies, animal studies, and case reports involving humans in which a stun gun or some other source of high voltage, low amperage direct current electric shocks were used to treat actual or simulated venomous bites or stings. We concluded that the use of stun guns or other sources of high voltage, low amperage direct current electric shocks to treat venomous bites and stings is not supported by the literature. PMID- 11434487 TI - The blight of the Bitterroot, the mysterious Rocky Mountain spotted fever, and the significant role of Wilson and Chowning--a commentary. PMID- 11434488 TI - The so-called "spotted fever" of the Rocky Mountains. 1902. PMID- 11434489 TI - Warnings in the wilderness. PMID- 11434490 TI - Evaluating and improving your classes. PMID- 11434491 TI - Seizures at high altitude in a patient on antiseizure medications. PMID- 11434492 TI - Disasters and therapeutic or prophylactic interventions. PMID- 11434493 TI - If you don't feel well at altitude... PMID- 11434494 TI - Mountaineering accidents in the European Alps: have the numbers increased in recent years? AB - OBJECTIVE: Media reports convey the impression that the incidence of fatal accidents in the European Alps has increased. Because more specific data are lacking, we analyzed available data from the mountain rescue services in Germany, Austria, southern Tirol, Zermatt/Switzerland, and Chamonix/France from 1987 until 1997. METHODS: Information was gathered from the annual reports of the Austrian Mountain Rescue Service, the Swiss Alpine Club Rescue Station in Zermatt, the Mountain Rescue Service of the Southern Tirol Alpine Club, the Mountain Rescue Service of the Bavarian Red Cross, and the Department of Mountain Medicine and Traumatology from the Hospital in Chamonix. RESULTS: Although the total number of rescue missions and injured alpinists increased significantly during the period, the number of fatalities retrieved during such rescue missions showed no significant increase. CONCLUSIONS: Even taking into account the varying definitions of "mountain accident" used in these countries, available data from the analyzed areas of the European Alps do not demonstrate a drastic increase in the number of fatalities. In the future, data concerning mountain accidents in the European Alps should be monitored according to standard definitions and stored by the International Commission for Alpine Rescue. PMID- 11434495 TI - Identification of risk factors for exertional heat-related illnesses in long distance cyclists: experience from the California AIDS Ride. AB - OBJECTIVES: To identify risk factors for exertional heat-related illnesses and to determine if patients who are afflicted with the human immunodeficiency virus or have acquired immunodeficiency syndrome have an increased risk of exertional heat related illness. DESIGN: Descriptive, retrospective, matched case-control study. SETTING: Mobile community of 2,650 participants in the California AIDS Ride 3. PARTICIPANTS: Participants included 117 patients presenting for medical care with heat-related illnesses and 234 age-, gender-, and registration site-matched control subjects who did not develop a heat-related illness. METHODS: Retrospective, matched case-control study utilizing univariate and multivariate conditional logistic regression to determine if human immunodeficiency virus seropositivity, the number of chronic medical illnesses, or the number of current medications known to affect heat dissipation increased the risk of exertional heat-related illnesses. RESULTS: The multivariate model revealed that patients with a greater number of chronic medical illnesses were at a significantly increased risk of a heat-related illness (odds ratio = 1.6, 95% CI = 1.2-2.1). The number of current medications (odds ratio = 1.1, 95% CI = 0.8-1.5) and human immunodeficiency virus seropositivity (odds ratio = 0.7, 95% CI = 0.2-3.1) were not significant predictors of risk for exertional heat-related illness. CONCLUSIONS: Those riders suffering from a greater number of chronic medical illnesses were at a greater risk for developing an exertional heat-related illness. Human immunodeficiency virus seropositivity alone was not associated with increased risk of exertional heat-related illness. PMID- 11434496 TI - Patient attitudes toward issues of environmental health. AB - OBJECTIVE: Although the place of environmental health in medical education has been cautiously explored, patient attitudes toward environmental issues have been largely ignored in the medical literature. The purpose of this study was to evaluate the baseline level of family practice patient concerns about a variety of environmental issues, including their major sources of information and their level of trust in each source. Patient desire for more environmental education was also assessed. METHODS: This cross-sectional, descriptive study was based on a confidential, 14 question survey administered to a convenience sample of 373 adult family medicine patients at 4 University of Wisconsin-Madison Department of Family Medicine affiliated clinics, including 2 urban, 1 suburban, and 1 rural site. Of 496 patients approached, 415 accepted, and 373 returned complete and acceptable questionnaires. The level of patient concern for each environmental issue, patient trust in each source of information, and the amount of education patients felt they had received from their physician were measured. RESULTS: A high level of concern for environmental issues existed among the study sample patients. Although only 18% of patients reported utilizing their physician as a major source of environmental information, physicians were considered the most trustworthy source. In general, patients felt that they had received an inadequate amount of environmental education from their physician regarding all environmental issues presented. CONCLUSIONS: Patients desire and would be receptive to more environmental education from their physicians. PMID- 11434497 TI - Centipede envenomation. AB - Five episodes of envenomation by centipedes in 2 patients are reported. These arthropods are fast-moving, frightening in appearance to some, and may display aggressive behavior. However, stings from these centipedes, like most found worldwide, caused no serious morbidity or mortality. Common effects included intense local pain, erythema, induration, and necrosis, as well as mild constitutional symptoms. All resolved without sequelae. Treatment included pain control, wound care, and tetanus immunization. PMID- 11434498 TI - Epidemiology of osteoporosis. PMID- 11434499 TI - Two cheers for P-values? AB - P-values are a practical success but a critical failure. Scientists the world over use them, but scarcely a statistician can be found to defend them. Bayesians in particular find them ridiculous, but even the modern frequentist has little time for them. In this essay, I consider what, if anything, might be said in their favour. PMID- 11434500 TI - Prevalence of urinary symptoms in urban Australian men aged 40-69. AB - BACKGROUND: This study was devised to determine the prevalence of urinary symptoms among men living in the Australian cities of Melbourne, Sydney or Perth, and to identify factors associated with the presence of moderate-to-severe urinary symptoms. METHODS: The study comprised a population-based sample of 1,216 men, aged 40-69 years, whose names were obtained through electoral rolls and who participated as controls in a case-control study of risk factors for prostate cancer. As part of a structured face-to-face interview, the men completed the International Prostate Symptom Score (IPSS). Men with moderate (IPSS = 8-19) or severe (IPSS > or = 20) urinary symptoms were compared with those with mild or no symptoms (IPSS < 8) using unconditional logistic regression. RESULTS: The age specific prevalence of moderate-to-severe urinary symptoms (IPSS > or = 8) in men aged 40-49, 50-59, 60-69 years was 16%, 23% and 28%, respectively. Compared with men with no or mild urinary symptoms (IPSS < 8), men with moderate-to-severe symptoms were more likely to report not currently living as married [odds ratio (OR) = 1.5; 95% confidence interval (CI) 1.1-2.0] and being circumcised (OR = 1.5; 95% Cl 1.2-2.0). The increased likelihood associated with drinking an average of > 60 g day(-1) of alcohol in the 2 years before interview was of marginal statistical significance (OR = 1.6; 1.0-2.6). There were no significant differences between men with IPSS > or = 8 and those with IPSS < 8 with respect to body mass index, education level, having had a vasectomy, or cigarette smoking. CONCLUSION: Among Australian men, being circumcised, or not currently living as married, were associated with increased prevalence of urinary symptoms. PMID- 11434501 TI - Interaction between two carcinogens in the two-stage clonal expansion model of carcinogenesis. AB - BACKGROUND: Exposure to two or more carcinogens may result in interactive effects in which the joint effect may be greater or less than that expected to arise as the sum of the effects of the two agents alone. In this article, we investigate the joint effects of exposure to two carcinogens within the context of the two stage clonal expansion model of carcinogenesis. METHODS: Different measures of interaction are considered based on the notions of response and dose additivity, and an index of synergy S due to Thomas (1982) used to broadly characterise the effects of joint exposure. RESULTS: Interactive effects based on the index S were found to be qualitatively similar, regardless of whether cancer risk was defined in terms of age-specific relative risk, or the cumulative probability of cancer occurrence at the same age. For joint exposure to two initiators or to two completers (affecting the first or second mutation rate in the two-mutation model, respectively), S assumed values near zero, reflecting an additive relative risk relationship. For joint exposure to two promoters (which increase the rate of proliferation of initiated cells that have sustained the first mutation), the relative-risk relationship was found to range from supramultiplicative (S > 1) in younger age groups, to subadditive (S < 0) in older ages. Other combinations of carcinogens involving promotion also displayed a broad range of interaction effects. CONCLUSIONS: These results differ markedly from those reported previously by Kodell et al. (1991) for an approximate form of the two-stage model, which predicts much higher values of the index of synergy S than the exact form of the model when promotion is involved. PMID- 11434502 TI - Transient or persistent asthma-like symptoms and lung growth over 2-year follow up in pre-adolescent children. AB - BACKGROUND: The main purpose of this study was to assess the effect of transient and continued asthma-like symptoms on lung function growth in preadolescent children. METHODS: The follow-up respiratory health survey has been conducted on the sample of 1,129 children aged 9 years over 2 years follow-up. The basic health end-points were the occurrence of asthma-like symptoms and slower lung function growth (SLFG), which was defined as the lung function gain over 2 years within the lowest quintile of the distribution of a given spirometric test. RESULTS: Adjusted odds ratios (OR) for SLFG [forced vital capacity (FVC)] were significantly higher only in the children having continued symptoms [OR = 3.39: 95% confidence interval (CI) = 1.39-8.27]. There was a consistent trend of adjusted ORs for SLFG [forced expiratory volume (FEV,) with the category of symptoms, where OR was 2.00 (95% Cl = 1.17-3.42) in children with transient symptoms: while 4.10 (95% Cl = 1.71-9.86) in children who had persistent symptoms. The corresponding ORs for SLFG [maximal mid-expiratory flow (FEF25_75c)] were 2.27 (95% Cl = 1.37-3.76); and 5.43 (95% Cl = 2.38-12.40). DISCUSSION: The association between asthma-like symptoms and lung function gain in preadolescent children confirmed the clinical significance of the symptoms in question. The observed slower lung function gain in preadolescence may have implications for the development of chronic lung disease later in adulthood. PMID- 11434503 TI - Estimation of disease progression parameters from case-control data: application to mammographic patterns and breast cancer natural history. AB - BACKGROUND: Estimations of mean sojourn time (MST) and sensitivity (S) in disease screening have been previously calculated from case-control data, using simple models which did not include covariates. Many studies have shown an effect of mammographic parenchymal pattern (MPP) on breast-cancer risk and tumour histology. We have expanded previous models on these to estimate MST and S with the effects of MPP as a covariate. METHODS: Data were from a nested case-control study within the East Anglian screening programme, with 875 cases and 2,601 controls. Estimates of disease progression and screening parameters were based on conditional likelihood calculation, using a Markov process model. Ninety-five per cent confidence intervals (CI) were calculated using the profile likelihood wherever possible and using a numerical estimate of the information matrix or the area under the likelihood curve where necessary. RESULTS: We obtained estimates of the incidence of preclinical disease, rate of transition from preclinical to clinical and screening sensitivity, and evaluated the association of these parameters with mammographic parenchymal pattern. A higher incidence of preclinical disease was found for high-risk MPP [relative incidence = 1.62 (95% CI: 0.89; 2.73)]. However, no difference in progression rate from preclinical to clinical disease between different MPP was found. Dense MPPs were associated with decreased sensitivity [relative sensitivity = 0.24 (95% CI: 0.06; 15)]. Wide CIs were found, probably being a consequence of the relative sparsity of interval cancer data. DISCUSSION: It is possible to estimate multiple parameters of disease progression and screening quality from case-control data. The reduction in sensitivity of the screening process associated with high-risk patterns presented here, could be of paramount interest for proposing new screening strategies, such as possible additional screening tools. PMID- 11434504 TI - Relationship between the Val158Met polymorphism of catechol O-methyl transferase and breast cancer. AB - A case-control study was performed to assess the potential influence of catechol O-methyl transferase (COMT) genotype on the risk of breast cancer in Korean women. One hundred and sixty-three histologically confirmed incident breast cancer cases and 163 age- and menopausal status-matched control individuals with no present or previous history of cancer were selected as study subjects. COMT genetic polymorphism was determined by gel electrophoresis after NlaIII enzyme digestion of amplified DNA. Odds ratios and 95% confidence intervals were estimated by unconditional logistic regression after adjustment for known or suspected risk factors of breast cancer. Women with at least one COMT lower enzyme activity associated allele (COMT-L) were at elevated risk for breast cancer (OR = 1.7, 95% CI = 1.04-2.78) compared with those homozygous for high enzyme activity associated COMT-H alleles. Among women with low (> or = 23.1) body mass index the COMT-L allele containing genotypes posed a marginally significant increased risk of breast cancer compared to the COMT-HH genotype (OR = 1.8, 95% CI = 0.95-3.48). Women with at least one COMT-L allele who had experienced a full-term pregnancy when aged over 30 years or were nulliparous had 2.7-fold increased risk; however, this increase did not reach statistical significance (OR = 2.7, 95% CI = 0.64-11.35). Furthermore, never-drinking and never-smoking women with at least one COMT-L allele were at increased risk of breast cancer compared to those with COMT-HH genotype with ORs of 2.0 (95% CI = 1.23-3.38) and 1.7 (95% CI = 1.04-2.62), respectively. These results are consistent with studies showing that COMT genotype of lower enzyme activity might be related to increase in risk of breast cancer, and extend this finding to Korean women. PMID- 11434505 TI - The effect of genetic polymorphism of cytochrome P450 CYP2C9 on phenytoin dose requirement. AB - The cytochrome P450 enzyme CYP2C9 catalyses the metabolism of numerous therapeutic agents, including the anti-epileptic drug phenytoin. CYP2C9 is genetically polymorphic: two allelic variants are known, CYP2C9*2 and CYP2C9*3, differing from the wild-type CYP2C9*1 by a single point mutation. Both mutant alleles are associated with markedly impaired metabolic capacity for many CYP2C9 substrates compared to the wild-type, resulting in raised serum drug levels upon a given dose. Because this may be relevant in treatment with phenytoin, we studied the effect of CYP2C9 genotype on phenytoin dose requirement in a group of 60 epileptic patients on long-term phenytoin therapy. CYP2C9 genotyping was performed by polymerase chain reaction analysis, phenytoin serum concentrations were measured by high-performance liquid chromatography analysis and related to the maintenance doses. For patients carrying at least one mutant CYP2C9 allele (n = 17), the mean phenytoin dose required to achieve a therapeutic serum concentration was about 37% lower than the mean dose required by wild-type individuals (199 mg/day versus 314 mg/day; P < 0.01). A low maintenance dose (< 200 mg/day) sufficed for 47% of carriers, while 58% of normals required a high dose (> 300 mg/day) for an effective serum level. The results show that there is a strong association between CYP2C9 allelic variants and phenytoin dose requirement. Since phenytoin has a narrow therapeutic index and genotyping may be carried out rapidly and at low cost, dosage adjustment based on CYP2C9 genotype, especially at the induction of therapy, would be of value in order to lower the risk of concentration dependent drug intoxications in carriers. PMID- 11434506 TI - The C3435T mutation in the human MDR1 gene is associated with altered efflux of the P-glycoprotein substrate rhodamine 123 from CD56+ natural killer cells. AB - P-glycoprotein (PGP) is a membrane protein which determines drug disposition in humans (e.g. digoxin). It is also expressed in various leukocyte lineages with highest expression in CD56+ natural killer cells. Recently, a polymorphism in exon 26 (C3435T) of this gene was shown to correlate with intestinal PGP expression and function in humans. Carriers homozygous for this polymorphism (TT) showed more than two-fold lower PGP expression and higher digoxin plasma concentrations compared to the CC group. However, it is not known whether this mutation in the MDR1 gene is also associated with altered PGP function in peripheral blood cells. We therefore assessed efflux of the PGP-substrate rhodamine 123 from CD56+ natural killer cells. Leukocytes were isolated from whole blood of 10 CC, 10 CT and 11 TT healthy Caucasian individuals. Using flow cytometry, rhodamine fluorescence was determined in CD56+ cells. Moreover, MDRI mRNA was quantified in leukocytes by real-time polymerase chain reaction. Subjects with CC genotype revealed a significantly lower rhodamine fluorescence (i.e. higher PGP function) compared to individuals with TT genotype (51.1 +/- 11.4% versus 67.5 +/- 9.5%, p < 0.01). Heterozygous individuals had an intermediate rhodamine fluorescence (61.4 +/- 6.3%). MDR1 mRNA normalized for cyclophilin was lowest in the TT population (1.29 +/- 1.01), intermediate in heterozygous subjects (1.60 +/- 0.76) and highest in the CC group (1.91 +/- 0.94; not significant). In summary, subjects being homozygous for C in position 3435 of the MDR1 gene have a more pronounced efflux of rhodamine from CD56+ natural killer cells and a higher MDR1 mRNA expression in leukocytes than subjects with the TT genotype. Measurement of rhodamine efflux using flow-cytometry from peripheral blood cells allows assessment of genetically determined differences in P-glycoprotein function. PMID- 11434507 TI - Uteroglobin gene polymorphisms affect the progression of immunoglobulin A nephropathy by modulating the level of uteroglobin expression. AB - Uteroglobin (UG) is an anti-inflammatory/immunomodulatory protein. Targeted disruption of UG rendered mouse glomerulonephritis resembling immunoglobulin (Ig)A nephropathy (IgAN). Sequence analysis on exon 1 of UG showed several putative binding sites for transcription factors, and polymorphisms in this site might influence the expression level of UG as a competitive protein. We speculated that the single nucleotide polymorphism at the 38th nucleotide (A to G) from the transcription initiation site of UG exon 1 would impact the progression of IgA nephropathy (IgAN). Polymerase chain reaction-restriction fragment length polymorphism and single-strand conformation polymorphism were instituted to determine the genetic polymorphism. Luciferase assay was performed using the gene constructs containing a region 404-bp long located upstream of UG exon 1 initiation site to analyse whether this polymorphism would affect the expression level. UG polymorphism was distributed no differently in patients with IgAN (n = 111) compared to 60 healthy control subjects. An excess of A genotype was found in one patient having progressive disease (P = 0.03) and the risk for the disease progression increased as the number of A alleles increased (P for trend = 0.03) after follow-up for 116 months. The odds ratio for progression with the AA genotype was 4.9 (95% Cl = 1.0-23.9) compared to patients having the GG genotype. Significant interactive effects of hypertension and genetic polymorphisms of UG on the disease progression were observed (P for interaction = 0.001). In the luciferase assay, the gene construct with A at the 38th site showed a decreased activity of 74 +/- 8.4% compared to that showed by G gene construct. Our results suggest that polymorphism at the 5' UTR region of UG exon 1 is an important marker for the progression of IgAN and may modulate the level of protein expression. PMID- 11434508 TI - Heterogeneity in hand veins responses to acetylcholine is not associated with polymorphisms in the G-protein beta3-subunit (C825T) and endothelial nitric oxide synthase (G894T) genes but with serum low density lipoprotein cholesterol. AB - Vascular responses to acetylcholine (ACh) are notoriously variable, the reason for this phenomenon is unknown. We tested the hypothesis that the variability in venous response to acetylcholine may be associated with two recently identified genetic polymorphisms for proteins involved in the signal transduction pathway, i.e. the G-protein beta3-subunit (GNB3) and endothelial nitric oxide synthase (eNOS). The dorsal hand vein technique was used in 37 healthy subjects. Hand veins were preconstricted with the alpha1-adrenoceptor agonist phenylephrine and the venodilator response to local ACh infusion was measured with and without comedication of acetylsalicylic acid or co-infusion of N(G)-monomethyl-L-arginine (L-NMMA). In addition, all subjects received routine laboratory tests and 26 of them were genotyped for the C825T polymorphism of the GNB3 gene and for the G894T polymorphism of the eNOS gene. A striking variability in venous response to ACh was found with dilation observed in the low ACh concentration range and reduced dilation or even constriction at high concentrations. ACh-induced venodilation was mediated by muscarinic receptors and abolished in the presence of both acetylsalicylic acid and L-NMMA suggesting dependence on endothelium. We did not find any association of the variability in ACh response with GNB3 or eNOS allele status. On the other hand, a significant positive correlation between ACh responsiveness and low density lipoprotein-cholesterol status was detected. Two recently discovered gene polymorphisms are not responsible for the profound heterogeneity in venodilator response to ACh. Surprisingly, this variability appears to relate to the lipid status of the subjects. The exact nature of this new finding requires further study. PMID- 11434509 TI - Nicotine metabolism and CYP2A6 allele frequencies in Koreans. AB - CYP2A6 is a major catalyst of nicotine metabolism to cotinine. Previously, we demonstrated that the interindividual difference in nicotine metabolism is related to a genetic polymorphism of the CYP2A6 gene in Japanese. To clarify the ethnic differences in nicotine metabolism and frequencies of CYP2A6 alleles, we studied nicotine metabolism and the CYP2A6 genotype in 209 Koreans. The cotinine/nicotine ratio of the plasma concentration 2 h after chewing one piece of nicotine gum was calculated as an index of nicotine metabolism. The genotypes of CYP2A6 gene (CYP2A6*1A, CYP2A6*1B, CYP2A6*2, CYP2A6*3, CYP2A6*4 and CYP2A6*5) were determined by polymerase chain reaction (PCR)-restriction fragment length polymorphism or allele specific (AS)-PCR. There were ethnic differences in the allele frequencies of CYP2A6*1A, CYP2A6*1B, CYP2A6*4 and CYP2A6*5 between Koreans (45.7%, 42.8%, 11.0% and 0.5%, respectively) and Japanese (42.4%, 37.5%, 20.1% and 0%, respectively, our previous data). Similar to the Japanese, no CYP2A6*2 and CYP2A6*3 alleles were found in Koreans. The homozygotes of the CYP2A6*4 allele (four subjects) were completely deficient in cotinine formation, being consistent with the data among Japanese. The heterozygotes of CYP2A6*4 tended to possess a lower metabolic ratio (CYP2A6*1A/CYP2A6*4, 4.79 +/- 3.17; CYP2A6*1B/CYP2A6*4, 7.43 +/- 4.97) than that in subjects without the allele (CYP2A6*1A/CYP2A6*1A, 7.42 +/- 6.56; CYP2A6*1A/CYP2A6*1B, 9.85 +/- 16.12; CYP2A6*1B/CYP2A6*1B, 11.33 +/- 9.33). The subjects who possess the CYP2A6*1B allele appeared to show higher capabilities of cotinine formation. It was confirmed that the interindividual difference in nicotine metabolism was closely related to the genetic polymorphism of CYP2A6. The probit plot of the metabolic ratios in Koreans (8.73 +/- 11.88) was shifted to a higher ratio than that in the Japanese (3.78 +/- 3.09). In each genotype group, the Korean subjects revealed significantly higher metabolic ratios than the Japanese subjects. The ethnic difference in cotinine formation might be due to environmental and/or diet factors as well as genetic factors. PMID- 11434510 TI - Relationship between glutathione S-transferase M1, P1 and T1 polymorphisms and early onset prostate cancer. AB - There is evidence suggesting that polymorphic variations in the glutathione S transferases (GSTs) are associated with cancer susceptibility. Inter-individual differences in cancer susceptibility may be mediated in part through polymorphic variability in the bioactivation and detoxification of carcinogens. The GSTs have been consistently implicated as cancer susceptibility genes in this context. The GST supergene family includes several loci with well characterized polymorphisms. Approximately 50% of the Caucasian population are homozygous for deletions in GSTM1 and approximately 20% are homozygous for deletions in GSTT1, resulting in conjugation deficiency of mutagenic electrophiles to glutathione. The GSTP1 gene has a polymorphism at codon 105 resulting in an Ile to Val substitution which consequently alters the enzymatic activity of the protein and this has been suggested as a putative high-risk genotype in various cancers. We investigated the relationship between GST polymorphisms and young onset prostate cancer in a case-control study. GSTM1, GSTT1 and GSTP1 genotypes were determined for 275 prostate cancer patients and for 280 geographically matched control subjects. We found no significant difference in the frequency of GSTM1 or GSTT1 null genotypes between cases and controls. GSTP1 genotype was, however, significantly associated with prostate cancer risk: the Ile/Ile homozygotes had the lowest risk and there was a trend in increasing the risk with the number of 105 Val alleles: Ile/Val odds ratio (OR)= 1.30 (95% FCI 0.99-1.69), Val/Val OR = 1.80 (95% FCI 1.11-2.91); Ptrend = 0.026. These results suggest that the GSTP1 polymorphism may be a risk factor for developing young onset prostate cancer. We also found that carrying more than one putative high-risk allele in the carcinogen metabolizing GST family was associated with an elevated risk for early onset prostate cancer (OR 2.48, 95% FCI 1.22-5.04, Ptrend = 0.017). PMID- 11434511 TI - Long sleep and short sleep mice differ in nicotine-stimulated 86Rb+ efflux and alpha4 nicotinic receptor subunit cDNA sequence. AB - In a recent study, we reported that a restriction fragment length polymorphism associated with the alpha4 nicotinic receptor gene (Chrna4) may play a role in regulating differential sensitivity of LS and SS mouse lines to the seizure inducing effects of nicotine. Since the alpha4 subunit (CHRNA4) is often found as a heteromer with the beta2 subunit (CHRNB2), alpha4 and beta2 cDNAs from the LS and SS mice were cloned and sequenced. A polymorphism in the coding portion of the alpha4 gene was found (1587A to G) which should result in a threonine/alanine substitution at position 529 (T529A). The LS and SS beta2 nicotinic receptor subunit cDNAs were identical. The potential consequences of the alpha4 polymorphism were evaluated using an ion (86Rb+) flux assay that likely measures the function of alpha4beta2-type receptors. LS-SS differences in maximal nicotine stimulated ion flux were seen when bovine serum albumin (BSA) was not included but this difference was not seen when BSA was included in the perfusion buffer. Current evidence suggests that BSA may alter the ratio of nicotinic receptors that are in the ground state and desensitized forms. Thus, it may be that the Chrna4 T529A substitution leads to a difference in the ratio of the two receptor forms which then promotes differences in receptor function, as well as differential behavioural sensitivity to nicotine. PMID- 11434512 TI - Effects of genotypic differences in CYP2C19 status on cure rates for Helicobacter pylori infection by dual therapy with rabeprazole plus amoxicillin. AB - Rabeprazole is a potent proton pump inhibitor and is mainly reduced to thioether rabeprazole by a non-enzymatic pathway and partially metabolized to demethylated rabeprazole by CYP2C19 in the liver. We intended to determine a cure rate for Helicobacter pylori infection by dual rabeprazole/amoxicillin therapy in relation to CYP2C19 genotype status prospectively. Ninety-seven patients with gastritis and H. pylori infection completed the dual therapy with 10 mg of rabeprazole bid and 500 mg of amoxicillin tid for 2 weeks. At 1 month after treatment, cure of H. pylori infection was assessed on the basis of histology, a rapid urease test, culture, polymerase chain reaction (PCR), and 13C-urea breath test. CYP2C19 genotype status was determined by a PCR-restriction fragment length polymorphism method. Of the 97 patients, 33 were homozygous extensive metabolizers (homEM), 48 were heterozygous extensive metabolizers (hetEM), and 16 were poor metabolizers (PM). Cure of H. pylori infection was achieved in 79 of the 97 patients (81.4%, 95%CI = 71.9-88.7). Significant differences in cure rates among the homEM, hetEM, and PM groups were observed; 60.6% (95%CI = 42.1-77.3), 91.7% (95%CI = 80.0 97.7), and 93.8% (95%CI = 69.8-99.8), respectively (P = 0.0007). Twelve patients without cure after initial treatment (10 homEMs and 2 hetEMs) were successfully retreated with rabeprazole 10 mg q.i.d. and amoxicillin 500 mg q.i.d. for 2 weeks. The cure rates for H. pylori infection by dual rabeprazole/amoxicillin therapy depended on the CYP2C19 genotype status. This dual therapy appears to be effective for hetEM and PM patients. However, high dose dual rabeprazole/amoxicillin therapy was effective even for homEM patients. Therefore, the genotyping test of CYP2C19 appears to be a clinically useful tool for the optimal dual treatment with rabeprazole plus amoxicillin. PMID- 11434513 TI - Use of midazolam urinary metabolic ratios for cytochrome P450 3A (CYP3A) phenotyping. AB - Midazolam (MDZ) total clearance (ClT) is widely used for cytochrome P450 3A (CYP3A) phenotyping, but requires up to eight blood samples. This study was conducted to compare the use of midazolam ClT to use of a midazolam urinary metabolic ratio for CYP3A phenotyping. Ten male and 10 female subjects received i.v. midazolam 0.025 mg/kg eight times over a 4-month period at approximately 2 week intervals. The first six phenotyping measures were used to estimate baseline CYP3A activity, then subjects received the moderate CYP3A inhibitor fluvoxamine 150 mg/day for the last 4 weeks (two phenotyping visits) of the study. Serial blood samples were obtained for calculation of ClT. Urine was collected for 6 h following each midazolam dose. Midazolam, 1'-hydroxymidazolam (1-OHMDZ), and 4 hydroxymidazolam were measured in plasma and urine by liquid chromatography with tandem mass spectrometry (LC/MS/MS). Analysis of 148 samples from 20 subjects revealed a weak overall correlation between the urinary ratio of 1-OHMDZ/MDZ to midazolam ClT of r(s) = 0.372 (P = 0.0001). There was no correlation when examining either baseline samples or fluvoxamine-inhibited samples alone (r(s) = 0.101, P = 0.289 and r(s) = 0.266, P = 0.123, respectively). The median (range) urinary ratio decreased significantly with fluvoxamine [219 (141-409) versus 127 (50-464); P = 0.005] and to a similar extent to the midazolam ClT (-33.6% versus 42.4%, respectively; P > 0.05). Median urinary recovery of the i.v. midazolam dose varied between 1.4% and 53% and was significantly lower in samples collected while patients were receiving fluvoxamine (34.3% versus 23.1%; P= 0.0004). Based on these results, although this midazolam urinary ratio was not very reflective of baseline CYP3A activity, it may be a useful indicator of CYP3A inhibition. PMID- 11434514 TI - Thirteen UDPglucuronosyltransferase genes are encoded at the human UGT1 gene complex locus. AB - The original novel UGT1 complex locus previously shown to encode six different UDP-glucuronosyltransferase (transferase) genes has been extended and demonstrated to specify a total of 13 isoforms. The genes are designated UGT1A1 through UGT1A13p with four pseudo ones. UGT1A2p and UGT1A11p through UGT1A13p have either nucleotide deletions or flawed TATA boxes and are therefore pseudo. In the 5' region of the locus, the 13 unique exons 1 are arranged in a tandem array with each having its own proximal TATA box element and, in turn, are linked to four common exons to allow for the independent transcriptional initiation to generate overlapping primary transcripts. Only the lead exon in the nine viable primary transcripts is predicted to undergo splicing to the four common exons generating mRNAs with identical 3' ends and transferase isozymes with an identical carboxyl terminus. The unique amino terminus specifies acceptor substrate selection, and the common carboxyl terminus apparently specifies the interaction with the common donor substrate, UDP-glucuronic acid. In the extended region, the viable TATA boxes are either A(A)TgA(AA)T or AT14AT; in the original locus the element for UGT1A1 is A(TA)7A and TAATT/CAA(A) for all of the other genes. UGT1A1 specifies the critically important bilirubin transferase isoform. The relationships of the exons 1 to each other are as follows: UGT1A2p through UGT1A5 comprises a cluster A that is 87-92% identical, and UGT1A7 through UGT1A13p comprises a cluster B that is 67-91% identical. For the two not included in a cluster, UGT1A1 is more identical to cluster A at 60-63%, whereas UGT1A6 is identical by between 48% and 56% to all other unique exons. The locus was expanded from 95 kb to 218 kb. Extensive probing of clones beyond 218 kb with coding nucleotides for a highly conserved amino acid sequence present in all transferases was unable to detect other exons 1. The mRNAs are differentially expressed in hepatic and extrahepatic tissues. This locus is indeed novel, indicating the least usage of exon sequences in specifying different transferase isozymes that have an expansive substrate range. PMID- 11434515 TI - Lipids in monogastric animal meat. AB - Meat from monogastric animals, essentially pigs and poultry, is from afar the most consumed of all meats. Meat products from every species have their own characteristics. For a long time, pig meat has been presented as a fatty meat because of the importance of subcutaneous adipose tissue. Actually, when the visible fat is separated, this meat is rather poor in lipids: pieces eaten as fresh meat and without transformation, such as roasts, contain less then 2% total lipids. Poultry meat has always had a reputation of leanness because of its low content in intramuscular lipids. In addition, adipose tissues, localised in the abdominal cavity, are easily separable. The progress in genetics and a better knowledge of dietary needs has allowed to improve growth performances, to increase muscle weight and, in the pig, to strongly decrease carcass adiposity. However, strong contradictions appear between transformers and nutritionists, especially concerning the pig: the former wish to have meat with adipose tissues containing a high percentage of saturated fatty acids and the latter wish meat with more unsaturated fatty acids. The consumer, however, regrets the pigs of yesteryear or the poultry bred on farmyard that had tastier meat. At the same time, however, they request meat with a low fat content, which is paradoxical. PMID- 11434516 TI - The "apports nutritionnels conseilles (ANC)" for the French population. AB - The apports nutritionnels conseilles (ANC) for the French population are the reference which are used in France to assess the nutritional status of the population. These references have been elaborated by a transparent and collective expert process, conferring on them an official reference. The originality of the French method was to add to the classical analysis of each nutrient an analysis concerning both a global approach to specific groups (children, pregnant women, elderly people, sportsmen) and the links between nutrients and foods. Many original works, which have been specifically performed for this revision, highlight the absolute need for rigorous evaluation of food consumption patterns and for valuable food composition tables. PMID- 11434517 TI - Forty years of achievement in French research on digestive physiology in the pig. AB - The contribution of the research group set up by Rerat in the early 1960s in Jouy en-Josas, and then expanded to Rennes, is summarised. As digestive processes are a major key to the nutritional status of monogastric animals, original methodologies based on advanced experimental surgery were developed in the pig to quantify the digestion and absorption yield, to know their factors of variation, and to understand their physiological and nutritional consequences. The group also extended its expertise to the control of food intake, the role of the nervous system and regulatory peptides, and several biomedical gut-related topics. PMID- 11434518 TI - Portal absorption of 15N and amino nitrogen in the growing pig after ingestion of labelled milk, yogurt or heat-treated yogurt. AB - The aim of this experiment was to study 15N and amino-nitrogen (AN) portal absorption in the growing pig after ingestion of uniformly (0.2509 APE) labelled 15N milk (M), yogurt ingested just after manufacturing (Y0), yogurt stored for 21 d at 4 degrees C (Y21) and heat-treated yogurt (HY). The highest porto-arterial differences (PAD) in 15N and AN were found in the period between 30 min and 90 min after ingestion. The absorption of nitrogen from M and HY mainly occurred during the 0-120 min time period (about 70% for M and 67% for HY). For Y0 and Y21, a larger displayed absorption period over the 0-240 min time period was observed. Y0 and Y21 presented a quite similar portal absorption profile. The 15N absorption rate was close to 80% for each studied milk product, suggesting that under our experimental conditions, dairy products (M, Y0, Y21 and HY) deliver nearly the same amounts of nitrogen to the organism. AN absorption rates were around 78% with a higher variability between the milk products. These results also indicate that most of the proteins were absorbed within the 240 min postprandial period. PMID- 11434519 TI - IGF-I, IGFBP-2 and -3 but not GH concentrations are different in normal and poor growing piglets. AB - In this study we investigated the somatotropic axis in piglets with evident growth delay. Female Suffolk crossbred piglets (30 days old; N = 12) were divided into normal weight (10 +/- 0.9 kg) and poor growing subjects (7 +/- 0.5 kg) and bled for growth hormone (GH), Insulin-like growth factor-I (IGF-I), Insulin-like growth factor binding protein 2 and 3 (IGFBP-2 and -3) determination. Basal and induced-GH levels were not different in the groups. Plasma IGF-I concentrations were significantly different (p < 0.001): 101.8 +/- 9.8 ng x mL(-1) (normal weight group) and 39.5 +/- 4.0 ng x mL(-1) (poor growing group). IGFBP-2 and -3 concentrations were significantly (p < 0.001) lower in poor growing than in normal piglets. Piglet weight was positively correlated (r = 0.98, p < 0.001) with IGF-I and IGFBP-2 or -3 concentrations. Our data indicate that growth rate was not correlated to basal or secretagogue-induced GH secretion. PMID- 11434520 TI - Sexual dimorphism for growth in Muscovy ducks and changes in insulin-like growth factor I (IGF-I), growth hormone (GH) and triiodothyronine (T3) plasma levels. AB - Muscovy ducks display marked sexual dimorphism for body weight. The aim of our study was to analyse the possible relationships between this dimorphism and plasma levels of T3, GH and IGF-I from hatch to 15 weeks of age. We found no significant effect of sex or age on plasma T3 which remained relatively stable about 6 nM x L(-1). Plasma GH levels were highest (14-24 ng x mL(-1)) but highly variable during the first four weeks in females and during the first seven weeks in males. Then, they decreased and remained low (3-4 ng x mL(-1)) until the age of 15 weeks. A trend for higher GH concentrations in males compared to females was observed across the experimental period. The difference was maximum around 6 7 weeks of age. Plasma IGF-I levels rose sharply between 2 and 3 weeks of age and remained high until 6-7 weeks of age for the females and 13 weeks of age for the males then started to decrease. For the whole experimental period, significantly higher IGF-I levels were measured in males. The differences between sexes were significant between 7 and 12 weeks of age and at 14 weeks of age. The differences in circulating GH and IGF-I levels between sexes suggest that the somatotrophic axis is implicated in Muscovy duck sexual dimorphism. PMID- 11434521 TI - Comparison of the effect of two different handling media on rabbit zygote developmental ability. AB - Despite the attention paid to culture media, the relevance of the handling medium at egg recovery/transfer is frequently overlooked. In the present work, we compare the effect of two different handling media (PBS and HEPES-buffered Ham F10, both supplemented with 20% (v/v) FCS), upon in vitro and in vivo developmental ability of in vivo fertilised rabbit zygotes. Zygotes recovered in HEPES-buffered medium (permanence 1 h as maximum) and subsequently cultured in vitro developed more efficiently to the compacted morula (100%) and blastocyst stage (92%) than those recovered in PBS (83% and 76%, respectively, P < 0.05). Zygotes recovered in such media were then further bilaterally transferred to recipient does following a brief in vitro culture period (for 4 hours). At caesarean section (day 28 of pregnancy), significant differences were observed in both the percentage of pregnant uterine horns (PBS: 60% vs. HEPES-buffered Ham F10: 100%) and live birth rates (PBS: 14% vs. HEPES-buffered Ham F10: 34%). Thus when early rabbit zygotes must be handled, even for short incubation periods, the medium is not innocuous. PMID- 11434522 TI - Fibrolytic activities and cellulolytic bacterial community structure in the solid and liquid phases of rumen contents. AB - Four sheep were fed an alfalfa hay diet. Rumen content samples were collected three hours after feeding in order to total microorganism population (TP), solid attached population (SAP) and solid attached firmly population (SAFP). Fibrolytic specific activities (xylanase, CMCase and beta-glycosidases) were estimated by the amount of reducing sugars or p-nitrophenol released from the appropriate substrate. The distribution of the three main cellulolytic bacterial species (Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefaciens) was quantified by dot-blot hybridisation using specific 16S-rRNA-targeting probes. Specific activities of polysaccharidase enzymes were higher in SAP than in TP, and in SAFP than in SAP. The sum of RNA of the three cellulolytic bacterial species represented on average 9% of the total bacterial RNA, and increased after filtration. In all samples, the relative population size of F. succinogenes was higher than that of R. albus and of R. flavefaciens. These results demonstrate that the most active enzymes are secreted by the particle-associated microorganisms. The differences in composition of the microflora between the solid and liquid phase suggest that bacteria are not equally distributed throughout the rumen content: the cellulolytic species are present in a higher proportion in the solid phase of rumen contents. PMID- 11434523 TI - Brucellosis control in Saudi Arabia: prospects and challenges. AB - Brucellosis is a zoonotic disease of worldwide distribution. Despite its control in many developed countries the disease remains endemic in Saudi Arabia where the national seroprevalence of the disease is 15%. In Saudi Arabia the disease is introduced through uncontrolled importation of animals that are poorly screened for the disease. Every year the Kingdom imports a few million heads of sheep and goats for sacrifice during Hajj from Africa, India, and Autstralia. Brucella melitensis remains the principle cause of human brucellosis in Saudi Arabia, causing 88-93% of the cases. Recent national statistics indicate that the disease incidence in humans is close to 40 cases per 100,000. The eradication of human brucellosis in Saudi Arabia will ultimately depend on the eradication of animal brucellosis. There is an urgent need for a national program for controlling brucellosis in the Kingdom. The components of this program will include recruitment and training of qualified veterinarians, development of an adequate number of animal quarantine centers and implementing legislation to control marketing and movement of animals. PMID- 11434524 TI - Trachoma: a review. AB - Trachoma is a leading cause of preventable blindness worldwide. The disease is caused by an intracellular epithelial gram-negative bacterium, Chlamydia trachomatis. The presence of children, overcrowding, and the lack of water in the household are factors that predispose to the transmission of the disease. The disease may remain asymptomatic but some patients many complain of redness, irritation, and ocular discharge. The principal initial clinical manifestation is a follicular conjunctivitis that may lead to conjunctival scarring, entropion, trichiasis, corneal thinning, and ulceration. Some patients develop corneal scars that lead to loss of vision. Despite the remarkable progress in our understanding of Chlamydial infections, the basic mechanisms involved in tissue damage and scarring remain to be elucidated. There are several effective therapeutic modalities for trachoma. Azithromycin oral single dose was found to be safe and effective in children with active trachoma. Conjunctival biopsy specimens obtained from adult patients receiving a single oral dose of azithromycin showed sustained high levels of azithromycin (above MIC of chlamydia) for up to 2 weeks after intake. These prolonged high levels of azithromycin in the conjunctival tissue following a single oral dose makes the drug suitable for the treatment of endemic trachoma. PMID- 11434525 TI - Community acquired pneumonia (CAP): an update. AB - Community-acquired pneumonia (CAP) remains a common and serious infectious disease worldwide. Recent investigations highlight the difficulties in making etiological diagnosis while emphasizing the importance of atypical pathogens in CAP. Prognostic tools such as the pneumonia severity index have been validated and widely endorsed as an aid in making site of care decisions in managing CAP patients. Empiric treatment guidelines for CAP have been developed and their use has been shown to simplify treatment regimens and improve outcome. PMID- 11434526 TI - Methicillin-resistant Staphylococcus aureus: profiles oceans apart--Canadian and Saudi Arabian experiences. AB - Methicillin-resistant Staphylococcus aureus (MRSA), a pathogen which has increased over the past three decades, is responsible for nosocomial infections and adverse patient outcomes. It is a pathogen of global importance. Rates of patient colonization or infection vary greatly internationally. Lower rates have been observed in Canada and Saudi Arabia compared to the United States and United Kingdom. Although these lower rates may appear reassuring, the trend in MRSA observed in two capital cities, Winnipeg, Canada and Riyadh, Saudi Arabia are consistent with a widespread global increase in MRSA. PMID- 11434527 TI - Bacterial meningitis in Saudi Arabia: the impact of Haemophilus influenzae type b vaccination. AB - Bacterial meningitis is one of the major causes of morbidity and mortality in children. A retrospective chart review of all cases of culture-proven bacterial meningitis in children was conducted in a tertiary care facility in the King Fahad National Guard Hospital (KFNGH), Riyadh. Sixty-seven patients with culture proven meningitis were reviewed. Bacterial meningitis is more common in children under 2 years of age (85%). Haemophilus influenzae type b (Hib) was the most common organism causing meningitis in children (57%). Streptococcus pneumoniae was the second most common organism (31%) followed by group B streptococcus in (7.5%). Fever, lethargy and vomiting were the most common presenting symptoms, occurring in 95%, 72%, and 66% respectively. The calculated incidence of Hib in KFNGH is 40/100,000. This incidence decreased dramatically after the initiation of routine infant vaccination in KFNGH with the conjugate Hib vaccine in April 1998. Outcome of Hib meningitis was good in 85% while outcome of Streptococcus pneumoniae was good in (53%). 43% of S. pneumoniae were resistant to penicillin. There was no cephalosporin-resistant isolate identified during the study period. Mortality due to meningitis was 4 (6%), 3 patients died due to S. pneumoniae and one due to Hib. Since Hib is the most common organism causing meningitis in Saudi Arabian children, mass vaccination of all Saudi children should be mandatory. PMID- 11434528 TI - Antibiotic resistance in developing countries. AB - During the past decade there have been major changes in the susceptibility of bacteria that cause various infections. Resistance to anti-infective agents, including antibiotics, is worldwide, both in developed and developing countries. Almost all bacterial species can develop resistance to anti-infective agents and resistance can readily be transferred among bacteria by transmissible elements (plasmids). Measures to prevent the emergence of resistance must be implemented urgently. A multiplicity of factors drive antibiotic resistance and solutions require the collaboration of governmental agencies, pharmaceutical companies, healthcare providers and consumers. Knowledge of resistance patterns and of the ways by which resistance is overcome is vital to the future of antimicrobial chemotherapy. PMID- 11434529 TI - Infection control practice: global preparedness for future challenges. AB - The modern infection control profession emerged in the 1960s in England and the US in response to a pandemic of infections in hospitalized patients from a virulent strain of Staphylococcus aureus. The need for better patient care practices to prevent infections in this setting was evident, and the nursing, medical and microbiology professions responded. Both governmental and non governmental agencies and professional organizations supported their efforts. During the past 3 decades infection control programs and professionals have multiplied and flourished throughout the world. At first, each country worked primarily within its own borders. In recent years collaboration among countries has increased dramatically, especially as related to education and training, research, and publishing. This article describes two examples of these partnerships. In the US, the Society for Healthcare Epidemiology of America (SHEA) and the Association for Professionals in Infection Control and Epidemiology, Inc (APIC) have joined with the King Fahad National Guard Hospital (KFNGH), a center-of-excellence for infection control in Riyadh, Saudi Arabia for educational meetings and infection control certification. The information exchanged and relationships developed through these activities will further efforts of both countries toward meeting infection prevention and control challenges of the future. PMID- 11434530 TI - Infectious diseases: career preparation. AB - The human resources for the discipline of Infectious Diseases are inadequate in many countries. There is no global definition of "Infectious Diseases" physicians and cover competency. Preparation for a career in this speciality varies greatly. In large populations in Asia and Africa, few individuals exist who have been trained to be Infectious Disease Clinicians. There is a great need by national and international societies to embrace this speciality and address the global deficiencies in this discipline by directing funding agencies as well as training institutions to redirect resources to strengthen the capacity of health professionals to deal with infectious diseases adequately throughout the world. PMID- 11434531 TI - Diagnostic value of Brucella ELISA IgG and IgM in bacteremic and non-bacteremic patients with brucellosis. AB - The diagnostic value of Brucella ELISA IgG and IgM has been evaluated in patients with brucellosis. Serum samples and blood cultures were collected from 83 patients with brucellosis. The sera were tested by Brucella ELISA for Brucella IgM and IgG antibodies. All 44 controls were negative for IgG and IgM. Brucella melitensis was isolated from blood cultures of 30/83 (36.1%) patients. Among the 30 bacteremic patients, 24 (80%) had an increased IgM titer of > or = 200. Of the 53 non-bacteremic patients, 41 had IgM titer > or = 200, while 22 had IgG titer of > or = 1,600. The ELISA IgM and IgG tests achieved a specificity and sensitivity of 100% and 96% respectively, while the positive and negative predictive values were 100% and 94% respectively. The Brucella ELISA is a reliable and sensitive test in the diagnosis of brucellosis. The test is rapid, easy to perform and can be automated. PMID- 11434532 TI - The correlation of agglutination titer with positive blood cultures in brucellosis: A comparison of two study periods. AB - Brucellosis is an endemic zoonosis in Saudi Arabia. Correlation of Brucella agglutination antibody titer with positive blood culture was performed over 2 time periods (1986 and 1998). At the King Fahad National Guard Hospital we generally consider a titer value of 1:320 or greater as being diagnostic of Brucella. Using this cut-off, the positive predictive value of 39.3% (95% CI=31.5 to 47.6) in the 1986 study increased to 55.7% (95% CI=42.4 to 68.5) in the recent study (P=0.03). This improvement is probably due to parallel improvement in both methodologies. PMID- 11434533 TI - The use of direct susceptibility tests from BACTEC 9240 to Microscan Walkaway. AB - Using a straight-from-the-bottle into-the-instrument technique, a series of direct susceptibility tests were performed with the BACTEC 9240 and Microscan Walkaway systems. Although rapid, unacceptably high error rates were obtained when compared with a standardized method. This procedure is not sufficiently accurate for clinical use. PMID- 11434534 TI - First case of combined blastomycosis and tuberculosis in a patient from Saudi Arabia. AB - Acute blastomycosis has not been reported on the Arabian Peninsula. We report an unusual case of dual pulmonary blastomycosis and tuberculosis. In addition, this is the first case of blastomycosis reported from Saudi Arabia, a disease not endemic in this region. PMID- 11434535 TI - Upper airway obstruction due to rhinoscleroma: case report. AB - Rhinoscleroma is a very rare cause of upper airway obstruction with only isolated reports in the literature of rhinoscleroma with isolated tracheal obstruction. The course is usually chronic with the presentation most often being non specific. We report a 54-year-old woman with progressive shortness of breath and wheezing over 7 years' duration. She was diagnosed and treated as bronchial asthma without improvement in her symptoms. At the time of referral to our institution, her flow-volume loop revealed fixed upper airway obstruction. Her chest radiography and other laboratory tests were normal. Bronchoscopy revealed a 70-80% irregular concentric stenosis of the trachea beginning immediately below the vocal cords and extending 4 cm distally. Biopsy showed characteristic Mikulicz histiocytes containing numerous gram-negative intracellular coccobacilli consistent with a diagnosis of rhinoscleroma. The patient was treated with laser resection of the stenosis followed by a course of ciprofloxcin and trimethoprim sulfamethoxazole. She has remained asymptomatic over a year follow-up period and repeated biopsies have shown no evidence of recurrence. PMID- 11434536 TI - Infectious diseases on the 21st century doorstep. AB - Accelerating change can be expected to occur in the fields of Clinical Microbiology and Infectious Diseases. New pathogens, new diseases, new technologies, new vaccines and new treatment modalities will descend on us with increasing frequency in the decade ahead. In order to cope with and even appreciate change, we need to increase our commitment to training, recruiting and retaining health professionals in these disciplines, enhance our research programs, and remain enthusiastic about the benefits offered by skilled, knowledgeable practitioners to our patients. Infectious disease deaths have been increasing during the '90's in most societies. We must reverse this! PMID- 11434537 TI - 18-Crown-6-Tetracarboxylic acid as a chiral additive for the simultaneous separation of o-, m- and p-enantiomers of phenylalanine family by capillary electrophoresis. AB - Simultaneous separation of 12 o-, m- and p-positional enantiomers of tyrosine and fluorophenylalanine has been achieved by the use of optically active 18-crown-6 tetracarboxylic acid (18C6H4) as an additive in the background electrolyte. It has been investigated that the separation conditions such as pH, electroosmotic flow, the concentration of 18C6H4 and applied voltages have critical influence on the simultaneous separation. Based on the information of electropherograms obtained, the interaction between the chiral selector and positional enantiomers has been discussed. It was shown that alpha-methyl group in the amino acid enantiomers make the resolution be reduced, and that o-, m- and p-substituents have an influence on the resolution. PMID- 11434538 TI - Synthesis and optical resolution property of (1 -->6)-alpha-D-glucopyranan tris(phenylcarbamate) as chiral stationary phase in HPLC. AB - The optical resolution property of 2,3,4-Tris-O-(3,5-dimethylphenylcarbamoyl)-(1- >6)-alpha-D-glucopyranan (5) toward racemic compounds has been studied as the chiral stationary phase (CSP) in high-performance liquid chromatography (HPLC). For the synthesis of (1-->6)-alpha-D-glucopyranan (4), the ring-opening polymerization of 1,6-anhydro-2,3,4-tri-O-allyl-beta-D-glucopyranose (1) has been proceeded through the stereoselective ring-opening mechanism to yield the 2,3,4 tri-O-allyl-(1-->6)-alpha-D-glucopyranan (2). For chromatographic enantioseparation of ten racemates, such as troger's base and benzoin, using 5 as a CSP, it was found that the chiral discrimination ability of 5 with the high molecular weight polymer (CSP-5a) was higher than that with the low molecular weight polymer (CSP-5b). In addition, the elution tendency of racemates using CSP 5a was essentially identical with the results of enantioseparation using dextran tris(3,5-dimethylphenylcarbamate). PMID- 11434539 TI - Synthesis and properties of optically active 1,1':5',1":5",1"'-hexol (QUANOL). AB - Optically active (R,R,R)- and (R,S,R)-1,1':5',1":5",1'"-quatemaphthalene 2,2',6',2",6",2"'-hexol (QUANOL) were prepared from (R)-BINOL in five steps. The CD spectrum of (R,R,R)-QUANOL showed a larger A value than those of (R)-BINOL and (R,S,R)-QUANOL, but additivity of the axial chiralities in the A value was not observed. Other basic properties for the use of QUANOL as a chiral ligand are also reported. PMID- 11434540 TI - Asymmetric synthesis of both enantiomers of point-difluorinated-eldanolide, analogues of insect sex pheromone. AB - Substitution of two fluorine atoms on organic molecules is expected to alter both chemical reactivity and biological activity due to the strong electron withdrawing nature of fluorine. The synthesis of partly gem-difluorinated compounds remains a significant challenge to synthetic organic chemists. We report that [2,3]-Wittig rearrangement of 1,1,2-trifluoroallylic ether gave a new type of partly gem-difluorinated allylic alcohol: 6-methyl-4,4,5-trifluorohept 1,5-dien-3-ol, 3, in a highly stereoselective fashion, and optical resolution of alcohol 3 was accomplished via lipase PS(PCL)-catalyzed reaction. Using this alcohol as the starting material, the first asymmetric synthesis of both enantiomers of point-fluorinated-eldanolides, 2,2,5,5,6-pentafluoroeldanolide 1 and 5,5,6-trifluoroeldanolide 2, analogues of the sex pheromone of the male African sugarcane borer, has been demonstrated. PMID- 11434541 TI - Enthalpy changes observed upon mixing liquid (R)- and (S)-enantiomers at 298.15 K. AB - Enthalpies of mixing of (R)- and (S)-enantomers of liquid chiral compounds such as 5-isopropenyl-2-methyl- 2-cyclohexanone (1), glycerol acetonide (2), 4 isopropenyl-1-methylcyclohexene(3), 2,6,6-trimethyl-bicyclo[3.1.1] hept-2-ene (4) have been measured over the whole range of mole fractions at 298.15 K, albeit very small values. Mixing of R-1 and S-1 realises enthalpic stabilisation over the whole range of mole fractions, whereas that of R-2 and S-2, R-3 and S-3, or R 4 and S-4 realise enthalpic destabilisation over entire compositions. The maximum values of enthalpies of mixing and the intermolecular interaction of bond dipoles obtained by the molecular mechanics calculations showed a linear correlation, except the compound having two chiral centers or a phenyl group. PMID- 11434542 TI - One-Pot preparation of optically active sec-alcohols, epoxides, and sulfoxides by a combination of synthesis and enantiomeric resolution with optically active hosts in a water suspension medium. AB - Optically active sec-alcohols, epoxides, and sulfoxides were prepared by one-pot process of a combination of synthesis and enantiomeric resolution through inclusion complexation with an optically active host in a water suspension medium. PMID- 11434543 TI - Preferential enrichment: an essential crystal structure. AB - The crystal structure of (+/-)-[2-[4-(2 hydroxyhexyloxy)phenylcarbamoyl]ethyl]dimethylsulfonium p-chlorobenzenesulfonate [(+/-)-SCC], which failed to effect preferential enrichment due to the chemical modification of the prototype (+/-)-[2-[4-(3-ethoxy-2- hydroxypropoxy)phenylcarbamoyl]ethyl]dimethylsulfonium p-chlorobenzenesulfonate [(+/-)-SC] showing preferential enrichment, has been determined by x-ray crystallographic analysis. The crystalline form of (+/-)-SCC is not a mixed crystal of the two enantiomers, but a racemic compound. In addition, the revised crystal structures of (+/-)-[2-[4-(3-ethoxy-2 hydroxypropoxy)phenylcarbamoyl]ethyl] dimethylsulfonium p-toluenesulfonate [(+/-) ST] and (+/-)-SC, both of which showed preferential enrichment, are described. The crystal structure of (+/-)-SCC is different from that of (+/-)-SC, but very similar to that of (+/-)-ST. The formation of the crystal phase of the (+/-)-SC type, which can be followed by polymorphic transition to afford the crystal one of (+/-)-ST type if necessary, seems to be essential for successful preferential enrichment. PMID- 11434544 TI - Current usage of nomenclature for parasitic diseases, with special reference to those involving arthropods. AB - Terminological confusion has been aggravated by efforts to develop a standardized nomenclature for parasitic diseases (SNOPAD) arising from the proposal by Kassai et al., 1988) for a standardized nomenclature of animal diseases (SNOAPAD). To restabilize international nomenclature of parasitic diseases it is recommended that, whenever appropriate, names should follow the 'International Nomenclature of Diseases' (IND) compiled by the Council for International Organizations for Medical Sciences (CIOMS/WHO, 1987). For diseases not included in IND, familiarity should guide the choice of name: traditional English language names of diseases should be preferred, e.g. 'malaria', 'scabies' or, for parasitic diseases having no traditional name, the taxonomic name of the causative organism should be applied, e.g. 'Brugia timori microfilaraemia'; 'Plasmodium malariae infection'; 'Simulium allergy'--instead of the generic derivatives proposed by SNOPAD, i.e. brugiosis, plasmodiosis and simuliidosis, respectively. For names of new diseases or those rarely mentioned, the suffix -osis would normally take precedence. Generally, the name of choice for any disease in any language should be the vernacular term, with commonest English usage preferred for international communication, and publications should include synonyms in the list of keywords. PMID- 11434545 TI - Laboratory and field evaluation of insect repellents as oviposition deterrents against the mosquito Aedes albopictus. AB - Three experimental approaches were used to evaluate the oviposition deterrency of three insect repellents, AI3-35765, AI3-37220 (piperidine compounds), and the standard N,N-diethyl-3-methylbenzamide (deet) to the mosquito Aedes albopictus Skuse (Diptera: Culicidae). Against laboratory-reared Ae. albopictus gravid females, the EC50 values of AI3-37220, AI3-35765 and deet were 0.004%, 0.008% and 0.011% in laboratory cages and 0.004%, 0.01% and 0.009% in an outdoor screened cage. For a natural population of Ae. albopictus tested in the field, the EC50 values were determined as 0.004%, 0.008% and 0.001%, respectively. Ageing concentrations of 0.1% of each repellent provided >50% effective oviposition deterrency against the laboratory population of Ae. albopictus for 13 days in laboratory cages, for 15 days in the outdoor cage, and for 21 days against field population of Ae. albopictus in Florida. These topical skin repellents are effective oviposition deterrents for Ae. albopictus when employed at relatively low application rates. PMID- 11434546 TI - Phlebotomine sandfly responses to carbon dioxide and human odour in the field. AB - Responses of Lutzomyia sandflies (Diptera: Psychodidae: Phlebotominae) to carbon dioxide (CO2) and human odour were investigated by field experiments in Parana State, southern Brazil. Catches of two predominant species: Lu. intermedia (Antunes & Coutinho) and Lu. whitmani Lutz & Neiva, were compared between traps baited with a human adult or with CO2 emitted at the human-equivalent rate. When the baits were only 40 cm apart, no difference of attractiveness was detected. When baits were separated by 20 m, however, significantly fewer sandflies (44% Lu. intermedia, 46% Lu. whitmani) were trapped with CO2 compared with human bait. This is the first field evidence that anthropophilic sandflies are attracted by human kairomones in addition to CO2. For both species [Lutzomyia intermedia and Lu. whitmani] [corrected], the proportion of human attractiveness attributable to CO2 was significantly more [corrected] for males than females; for Lu. intermedia males human bait was no more attractive than CO2 alone. Gender differences in sandfly olfactory sensitivity are likely to be associated with behavioural differences on the host, where females feed on blood and males find mates. With traps 20 m apart, both Lutzomyia spp. showed roughly linear increased responses (log-log scale) to 0.08-0.55% CO2 equivalent to 0.5-4 humans. This would explain why host size is generally proportional to attractiveness, as observed for other species of phlebotomine sandflies. PMID- 11434547 TI - Transovarial transmission of African swine fever virus in the argasid tick Ornithodoros moubata. AB - The aim of this study was to determine filial infection prevalence of experimentally infected colony Ornithodoros moubata Walton (Ixodoidea: Argasidae) ticks for African swine fever virus (ASFV). Three groups of ticks were used: an uninfected control group, one group orally infected with the VIC T90/1 isolate and another group orally infected with the LIV 13/33 isolate of ASFV. The results show that filial infection prevalences were not constant but were highly variable between egg batches from different ticks and between successive egg batches from the same tick. Filial infection prevalences ranged from 1.8% to 31.8% for ticks infected with the VICT90/1 isolate and from 1.2% to 35.5% for ticks infected with the LIV 13/33 isolate. A similar pattern was noted after the third feed. Immunohistochemisty showed that virus replicates in the developing larval cells and not in the yolk sac cells or within the outer layers of the eggs. The results show that ASFV can replicate to a high titre (10(5.1)log10HAD50) within the larval cells of the developing egg. PMID- 11434548 TI - Field studies on efficacy of host odour baits for the biting midge Culicoides impunctatus in Scotland. AB - The efficacy of some putative attractants for the biting midge Culicoides impunctatus (Goetghebuer) (Diptera: Ceratopogonidae) was assessed using odour baited 'delta traps' and suction traps. 1-octen-3-ol was confirmed as a potent olfactory attractant for C. impunctatus when released at 0.06mg/h. Acetone (23mg/h) and a mix of six phenolic compounds (phenol, 3-ethylphenol, 4 ethylphenol, 3-methylphenol, 4-methylphenol and 4-propylphenol), at undetermined release rate, also significantly increased delta trap catches compared to unbaited controls. When tested in combination, there was evidence of synergism between CO2 (0.2L/min) and acetone, 1-octen-3-ol or cow urine, trap catches being, respectively, 4.7, 6.2 and 9.3-fold greater than for CO2 alone. Highest catches were obtained with triple bait combinations comprising cow urine + acetone + CO2 or cow urine + 1-octen-3-ol+CO2, which increased trap catches by X 22 and X 24, respectively, compared to CO2 alone. Culicoides impunctatus was found to be extremely sensitive to CO2 and responses, gauged over two field seasons, showed a significant dose-dependent increase in catch across the entire range of release rates (0.2-2.5 L/min). Responses to these release rates, ranging from small to large mammal equivalents, emphasized the important role of CO2 in host location by C. impunctatus. Uses of olfactory attractants for monitoring and control of Culicoides are reviewed on the basis of these results. PMID- 11434549 TI - Abiotic factors influencing the ecology of wild rabbit fleas in north-eastern Spain. AB - During 1992, the population dynamics of rabbit fleas were compared at two sites in north-eastern Spain. The sites differed mainly in terms of annual rainfall and soil type. All flea species showed seasonal cycles of abundance, although peaks in numbers occurred at different times, reflecting their specific adaptations for coping with climatic variables. Adult Spilopsyllus cuniculi (Dale) (Siphonaptera: Pulicidae) were found largely parasitizing rabbits in spring and adult Caenopsylla laptevi (Beaucournu etal.) (Siphonaptera: Ceratophyllidae) in the autumn. In contrast, monthly flea indices of Xenopsylla cunicularis (Smit) (Siphonaptera: Pulicidae) and Echidnophaga iberica (Ribeiro et al.) (Siphonaptera: Pulicidae) peaked in summer. Spilopsyllus cuniculi was present at both sites, but was less common on the drier site, where monthly mean temperature and annual rainfall approached the flea's physiological limits. By contrast, E. iberica, X. cunicularis and C. laptevi, known to be better adapted for dryness, showed the opposite patterns of abundance. Nevertheless, even these arid-adapted species took advantage of the milder and wetter spring (X. cunicularis and E. iberica) or autumn (C. laptevi) for breeding and larval development. Although environmental temperature, rainfall and soil texture will influence the microclimate of the burrows where the flea larvae develop, burrow humidity seems to be more dependent on soil characteristics and past rainfall rather than the humidity of the external air. PMID- 11434550 TI - Ambient temperature effects on the extrinsic incubation period of Wuchereria bancrofti in Aedes polynesiensis: implications for filariasis transmission dynamics and distribution in French Polynesia. AB - Temperature effects on development of the human filarial parasite Wuchereria bancrofti (Cobbold) (Filaridea: Onchocercidae) in the main Pacific vector Aedes polynesiensis Marks (Diptera: Culicidae) are analysed in relation to ambient climatic conditions. A statistical model of the extrinsic cycle duration as a function of temperature is described and used to distinguish three patterns of W. bancrofti transmission dynamics: continuous, fluctuating and discontinuous, occurring from north to south geographically among French Polynesian archipelagos. In the northerly Marquesas Islands (8-11 degrees S) filariasis transmission is continuous and very active, facilitated by perennially high temperatures combined with constantly high rates of man-vector contact. In the southerly Australes Islands (21-28 degrees S) filariasis transmission is seasonally discontinuous and, during the cooler months (May-September), the model predicts virtually no transmission because the cycle duration exceeds the life expectancy of the vector. In the Society Islands (16-18 degrees S), between the Marquesas and Australes, transmission is predicted to be intermediate as expected from their latitude, with seasonally fluctuating transmission potential. In the Tuamotu Islands (also geographically intermediate: 14-23 degrees S), with theoretically perennial transmission potential, transmission occurs only intermittently, being limited by other human and environmental factors whereby man-vector contact is confined to seasonal agricultural situations. Generally, among French Polynesian archipelagos where Aedes polynesiensis is the vector, the transmission potential for W. bancrofti and resulting disease manifestations of lymphatic filariasis in humans are correlated with ambient temperature due to the degree of southern latitude. PMID- 11434551 TI - Rate of development of Hydrotaea rostrata under summer and winter (cyclic and constant) temperature regimes. AB - The black carrion fly Hydrotaea rostrata Robineau-Desvoidy (Diptera: Muscidae) is a muscid, which occurs on approximately one-third of decomposing human corpses involved in homicide cases in south-western Australia. Work to date on its development rates is scant with only one published source available. The current study measured the precise developmental rates of this species using high repetition and sampling rates. A comparison was made of the developmental rates between constant and cyclic temperatures in winter and summer temperature regimes in south-western Australia. Developmental times for 90% of first stage larvae to emerge as adult flies are: summer constant, 21.6days, summer cyclic, 23.5 days, winter constant 64.5days and winter cyclic, 48.3 days. These data will allow forensic entomologists to make more accurate determinations of post-mortem intervals in cases where H. rostrata life history stages are present. PMID- 11434552 TI - Characterization of the screwworm flies Cochliomyia hominivorax and Cochliomyia macellaria by PCR-RFLP of mitochondrial DNA. AB - The primary screwworm fly Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae) is one of the most important insect pests of livestock in neotropical regions, whereas Cochliomyia macellaria (Fabricius) (Diptera: Calliphoridae), the secondary screwworm, is of medical and sanitary importance because of its role in the dissemination of pathogens. These two species share morphological similarities and both may occur in the same myiasis, but in different developmental stages. In this work, the usefulness of PCR-RFLP (polymerase chain reaction - restriction fragment length polymorphism) of mitochondrial DNA (mtDNA) for the unambiguous identification of C. hominivorax and C. macellaria was investigated. Two specific regions of mtDNA were amplified: 870bp from Cytochrome oxidase subunit I and 2100bp from the A+T rich/12S region from C. hominivorax and C. macellaria specimens from different areas of Brazil. Reliable species-specific PCR-RFLP results were obtained for the CO I region and the A+T rich/12S region using the restriction enzymes Dra I and Ssp I. These results confirm the conservation of CO I diagnostic restriction sites previously reported and demonstrate the usefulness of the control region sequences as an efficient marker for PCR-RFLP identification of Brazilian screwworm flies. The occurrences of intraspecific polymorphic patterns are discussed based on frequencies and potential conflicts for species identification. PCR-RFLP provides a potentially useful method for identifying samples from the areas where these species are monitored. PMID- 11434553 TI - Comparative fecundity and survival rates of phlebotomus papatasi sandflies membrane fed on blood from eight mammal species. AB - Female sandflies, Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae), were fed via chicken membrane on heparinized blood from eight species of mammal (human, horse, cow, pig, dog, rabbit, guinea-pig, hamster) and their reproductive success rates were compared. No appreciable differences between those fed on human and animal blood were detected with respect to the proportion of flies that fed successfully, mortality-rate within 24h, number of eggs laid per blood-fed female or egg viability. When mass-rearing sandflies for research purposes, membrane feeding avoids practical difficulties encountered if sandflies are allowed to feed on live hosts (i.e. anaesthesia, distress from handling and postfeeding inflammation) and reduction of sandfly fecundity due to host antibody interference. Use of animal blood also eliminates risks of accidental transmission of human blood-borne pathogens, e.g. hepatitis B and human immunodeficiency virus (HIV), and is less expensive than maintenance of animals and their preparation for sandfly feeding. PMID- 11434554 TI - Olfactory memory in the mosquito Culex quinquefasciatus. AB - The cosmotropical urban mosquito Culex quinquefasciatus Say (Diptera: Culicidae) uses chemical cues to locate suitable water pools for oviposition. Although gravid females are innately attracted to or repelled by certain compounds, this study found that an individual mosquito's preferences for these odours could be altered greatly by prior experience. Mosquitoes reared in water containing skatole, at a level normally repellent to ovipositing females, preferred to oviposit in water containing that compound rather than in water with an otherwise attractive odour compound (P-cresol). This behaviour occurred regardless of whether mosquitoes were tested individually or in groups of up to 50 per cage. The F1 progeny of conditioned mosquitoes did not exhibit the parental preference, but were as susceptible to conditioning as their parents. Moreover, rearing mosquitoes in infusions of hay or animal (guinea-pig) faeces produced a similar although less dramatic change, such that the innate propensity for hay infusion could be cancelled by rearing in guinea-pig faeces infusion. The results demonstrated a change in odour preference by Cx. quinquefasciatus following exposure to the odour during development or pupal eclosion, suggesting that some form of larval conditioning or early adult imprinting occurred. Precisely when that conditioning occurred remains to be determined. PMID- 11434555 TI - Seasonal changes in prevalence and intensity of Hypoderma actaeon in Cervus elaphus from central Spain. AB - Hypodermosis in red deer, Cervus elaphus Linnaeus (Artilodactyla: Cervidae), caused by the warble fly, Hypoderma actaeon Latreille (Diptera: Oestridae), was studied at Quintos de Mora (Toledo, Spain) over a one-year period between October 1994 and September 1995. One hundred and twenty-five red deer were examined for the presence of warble fly larvae. The prevalence of warbles was 44.8% and the intensity of infection was 38.29 (SD +/- 61.32) warbles/deer infected. Clear seasonal variations were observed in prevalence and intensity, with the highest values in the autumn and winter. Statistically significant differences were observed in prevalence and intensity among age groups. In contrast, no differences were found in prevalence and intensity between males and females. PMID- 11434556 TI - Diptera as vectors of mycobacterial infections in cattle and pigs. AB - Mycobacteria were isolated from 14 (4.5%) of 314 samples, containing 7791 adult Diptera, which were collected in the Czech Republic and Slovakia in 1997-2000. These flies were collected from three cattle herds with paratuberculosis, two pig herds with mycobacterial infections and one farm that kept both cattle and pigs and that did not have problems of mycobacterial infections. Mycobacterium intracellulare was isolated from Eristalis tenax Linnaeus (Diptera: Syrphidae) captured from a pig herd. Mycobacterium avium ssp. avium (serotype 8) was isolated from flies of the genera Drosophila Fallen (Diptera: Drosophilidae) and Musca Linnaeus (Diptera: Muscidae) originating from a pig herd. Mycobacterium spp. were isolated from Musca spp. and Mycobacterium fortuitum was isolated from dung flies of the genus Scatophaga Meigen (Diptera: Scatophagidae), Musca spp. and Stomoxys calcitrans Linnaeus (Diptera: Muscidae) captured in the same herd. Mycobacterium scrofulaceum was isolated from S. calcitrans from the farm with both cattle and pigs. Mycobacterium avium ssp. paratuberculosis was isolated from Scatophaga spp. collected from pastures grazed by one of the cattle herds and from Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae) and Lucilia caesar Linnaeus (Diptera: Calliphoridae) captured in a slaughterhouse, where cattle infected with paratuberculosis were slaughtered. Mycobacterium phlei was isolated from flies of the genus Lucilia captured at a waste bin. These data indicate that mycobacteria may be spread by adult flies that have been in contact with material contaminated with these pathogens. PMID- 11434557 TI - Establishment of the cat flea (Ctenocephalides felis felis) on the ferret (Mustela putorius furo) and its control with imidacloprid. AB - As the ferret, Mustela putorius furo L. (Carnivora: Mustelidae), is becoming increasingly popular as a pet animal and as it is susceptible to the cat-flea, Ctenocephalides felis felis Bouche (Siphonaptera: Pulicidae), an experimental model was established for evaluating insecticidal treatments on this host. A high establishment rate (76.7-91.8%) was recorded when 60 unfed adult C. felis were placed on ferrets. This provided an adequate infestation for chemotherapeutic evaluation without causing undue discomfort to the host. Twelve ferrets were allocated to two groups matched for sex and individual ability to sustain a flea population. One group was treated topically with an imidacloprid spot-on formulation at a dose rate of 10 mg/kg body-weight on Day 0. All ferrets were infested with C. felis on Days -1, 7, 14, 21 and 28, and flea counts were performed 8 and 24 h post-treatment and one day after each subsequent infestation. Fleas were removed at all but the 8 h count (when they were returned to their host). Flea burdens were reduced by 95.3% (P < 0.001) within 8 h of treatment and 100% efficacy was recorded at 24 h. At 1, 2, 3 and 4 weeks post treatment, protection against re-infestation was 92.9% (P < 0.001), 55.7% (P < 0.02), 18.3% (NS) and 7.4% (NS), respectively. Thus, at this dose rate, imidacloprid gave excellent efficacy against a resident C. felis population and provided a high level of residual activity for at least one week after treatment. PMID- 11434558 TI - Egg hatching of mosquitoes Aedes caspius and Ae. vittatus stimulated by water vibrations. AB - Simple laboratory experiments with eggs of wild-caught Aedes caspius (Pallas) and Aedes vittatus (Bigot) mosquitoes (Diptera: Culicidae) demonstrated that egg hatch rates (i.e. percentage hatching daily) increased significantly in response to water vibrations caused by finger-tip drumming on containers of submerged eggs. When disturbed by daily vibrations for only 30 s, eggs hatched sooner than when they remained flooded or were periodically drained and reflooded without agitation. For autogenous Ae. caspius from saltmarsh, egg hatch rates were three- to four-fold greater for groups of 50 eggs than for solitary eggs, possibly due to chemicals (functional kairomones) emanating from other eggs or larvae. For anautogenous Ae. vittatus from freshwater rock-pools, tapping the container daily (for 30 s each 2 h in 8 h) hatched 93% of eggs, compared with only 42% hatch of eggs agitated only when first flooded or 55% hatch of eggs dried and reflooded three times during 17 days without other disturbance. It is concluded that, apart from flooding and other factors, vibrations - simulating the patter of rainfall provide a very significant hatching stimulus for Aedes eggs. PMID- 11434559 TI - Sex chromosome variation and cytotaxonomy of the onchocerciasis vector Simulium squamosum in Cameroon and Nigeria. AB - On the basis of sex chromosome variation, three cytotypes of Simulium squamosum (Enderlein) (Diptera: Simuliidae) are described from Cameroon and Nigeria. Simulium squamosum A is the typical form as originally described by Vajime & Dunbar (1975) with chromosome I as the sex chromosome. It occurs throughout most of Cameroon and south-east Nigeria. A second cytotype, S. squamosum B, is described from the river Sanaga (Cameroon). It also has chromosome I as the sex chromosome, but the nature of the sex differential region is different. Simulium squamosum C has no sex-linked chromosomal rearrangements. It is widespread in Nigeria and occurs near Mount Cameroon, where it seems to hybridize with S. squamosum A. PMID- 11434560 TI - Dipsticks for rapid detection of plasmodium in vectoring anopheles mosquitoes. AB - Malaria remains the most serious vector-borne disease, affecting some 300-500 million people annually, transmitted by many species of Anopheles mosquitoes (Diptera: Culicidae). Monoclonal antibodies developed against specific circumsporozoite (CS) proteins of the main malaria parasites Plasmodium falciparum and P. vivax have been used previously for enzyme-linked immunosorbent assays (ELISA), widely employed for detection of malaria sporozoites in vector Anopheles for local risk assessment, epidemiological studies and targeting vector control. However, ELISA procedures are relatively slow and impractical for field use. To circumvent this, we developed rapid wicking assays that identify the presence or absence of specific peptide epitopes of CS protein of the most important P. falciparum and two strains (variants 210 and 247) of the more widespread P. vivax. The resulting assay is a rapid, one-step procedure using a 'dipstick' wicking test strip. In laboratory assessment, dipsticks identified 1 ng/ mL of any of these three CS protein antigens, with sensitivity nearly equal to the CS standard ELISA. We have developed and are evaluating a combined panel assay that will be both qualitative and quantitative. This quick and easy dipstick test (VecTest Malaria) offers practical advantages for field workers needing to make rapid surveys of malaria vectors. PMID- 11434561 TI - Redox regulation in human melanocytes and melanoma. AB - The human melanocyte is continuously exposed to intrinsic and extrinsic sources of reactive biochemical species, but is finely tuned via the intrinsic anti oxidant and radical properties of melanin to suppress the build-up of an altered redox phenotype. We propose that this control is lost during melanomagenesis and inappropriate redox-sensitive transcriptional factor activations occur which result in enhancement of an anti-apoptotic phenotype in the transformed cell. This conceptual framework offers testable steps to determine the role of redox alterations in the carcinogenic evolution, prevention and treatment of melanoma and other diseases of the melanocyte. PMID- 11434562 TI - Approaches to increasing skin melanin with MSH analogs and synthetic melanins. AB - Increased public awareness of ultraviolet (UV) radiation-induced skin cancers has lead to new interest in technologies for protection from sun exposure. Although many sun protection formulations are available, few of them attempt to achieve that provided by melanin itself, i.e., wide-spectrum absorbance of radiant energy coupled with anti-oxidant activity from a single product. In that regard, technologies in two separate areas are at or near the commercialization stage: 1) hormonal enhancement of natural skin melanin content, and 2) inclusion of natural and synthetic melanins in cosmetic formulations to impart melanin-like color to the skin. In this article, these approaches are briefly summarized using as examples Melanotans I and II, superpotent analogs of the melanin-stimulating hormone melanocortin (MSH), and Melasyn, a group of plant-derived synthetic melanins that have successfully been incorporated into cosmetic formulations for use as sun protectants and as cover-ups for problems resulting from uneven pigmentation, such as seen in vitiligo. PMID- 11434563 TI - EDNRB/EDN3 and Hirschsprung disease type II. AB - The study of vertebrate pigmentary anomalies has greatly improved our understanding of melanocyte biology. One such disorder, Waardenburg syndrome (WS), is a mendelian trait characterized by hypopigmentation and sensorineural deafness. It is commonly subdivided into four types (WS1-4), defined by the presence or absence of additional symptoms. WS type 4 (WS4), or Shah-Waardenburg syndrome, is also known as Hirschsprung disease Type II (HSCR II) and is characterized by an absence of epidermal melanocytes and enteric ganglia. Mutations in the genes encoding the endothelin type-B receptor (EDNRB) and its physiological ligand endothelin 3 (EDN3) are now known to account for the majority of HSCR II patients. Null mutations in the mouse genes Ednrb and Edn3 have identified a key role for this pathway in the normal development of melanocytes and other neural crest-derived lineages. The pleiotropic effects of genes in this pathway, on melanocyte and enteric neuron development, have been clarified by the embryologic identification of their common neural crest (NC) ancestry. EDNRB and EDN3 are transiently expressed in crest-derived melanoblast and neuroblast precursors, and in the surrounding mesenchymal cells, respectively. The influence of EDNRB-mediated signaling on the emigration, migration, proliferation, and differentiation of melanocyte and enteric neuron precursors, in vivo and in vitro has recently been the subject of great scrutiny. A major emergent theme is that EDN3-induced signaling prevents the premature differentiation of melanocyte and enteric nervous system precursors and is essential between 10 and 12.5 days post-coitum. We review the present understanding of pigment cell development in the context of EDNRB/EDN3--a receptor-mediated pathway with pleiotropic effects. PMID- 11434564 TI - Interaction of melanin with proteins--the importance of an acidic intramelanosomal pH. AB - Melanin is a highly irregular heteropolymer consisting of monomeric units derived from the enzymatic oxidation of the amino acid tyrosine. The process of melanin formation takes place in specialized acidic organelles (melanosomes) in melanocytes. The process of melanin polymerization requires an alkaline pH in vitro, and therefore, the purpose of an acidic environment in vivo remains a mystery. It is known that melanin is always bound to protein in vivo. It is also seen that polymerization in vitro at an acidic pH necessarily requires the presence of proteins. The effect of various model proteins on melanin synthesis and their interaction with melanin was studied. It was seen that many proteins could increase melanin synthesis at an acidic pH, and that different proteins resulted in the formation of different states of melanin, i.e., a precipitate or a soluble, protein-bound form. We also present evidence to show that soluble protein-bound melanin is present in vivo (in B16 cells as well as in B16 melanoma tissue). An acidic pH appeared to be necessary to ensure the formation of a uniform, very high molecular weight melano-protein complex. The interaction between melanin and proteins appears to be largely charge-dependent as evidenced by zeta potential measurements, and this interaction is also increased in an acidic pH. Thus, it appears that an acidic intramelanosomal pH is essential to ensure maximum interaction between protein and melanin, and also to ensure that all the melanin formed is protein-bound. PMID- 11434565 TI - Cyclic oscillations in melanin composition within hairs of baboons. AB - The wild-type agouti-banding pattern for hair is well characterized in lower mammals such as mice. The switch between eumelanin and pheomelanin in bands in the hair results from the interaction of alpha-melanocyte stimulating hormone and agouti signal protein through the melanocortin 1 receptor on melanocytes. However, such banding patterns have not been described to date in higher mammals. We now report such 'agouti'-banding patterns that occur in several subspecies of baboons, and characterize those hairs using chemical and immunohistochemical methods. Hair and skin samples were obtained from the dorsa of adult male baboons of different subspecies (Papio cynocephalus hamadryas (PCH) and Papio cynocephalus anubis (PCA)). The hairs were excised with scissors into the gray and the white bands of the PCH subspecies and into the black and the yellow bands of the PCA subspecies, and were analyzed for total melanin, eumelanin, and pheomelanin by spectrophotometric and chemical methods. Hairs in the PCA subspecies oscillate between a eumelanic band (with high melanin content) and a pheomelanic band, while hairs in the PCH subspecies oscillate between a eumelanic band (with low melanin content) and a non-pigmented band. Those chemical data are consistent with the histological appearance of the hair bulbs stained by the Fontana-Masson technique. The difference in the melanin content between PCH and PCA subspecies is most likely related to tyrosinase levels, as suggested by the presence of unpigmented muzzle in the PCH subspecies compared with the black muzzle in the PCA subspecies. PMID- 11434566 TI - Inhibition of melanosome transfer from melanocytes to keratinocytes by lectins and neoglycoproteins in an in vitro model system. AB - We propose that some of the critical molecules involved in the transfer of melanosomes from melanocytes to keratinocytes include plasma membrane lectins and their glycoconjugates. To investigate this mechanism, co-cultures of human melanocytes and keratinocytes derived from neonatal foreskins were established. The process of melanosome transfer was assessed by two experimental procedures. The first involved labeling melanocyte cultures with the fluorochrome CFDA. Labeled melanocytes were subsequently co-cultured with keratinocytes, and the transfer of fluorochrome assessed visually by confocal microscopy and quantitatively by flow cytometry. The second investigative approach involved co culturing melanocytes with keratinocytes, and processing the co-cultures after 3 days for electron microscopy to quantitate the numbers of melanosomes in keratinocytes. Results from these experimental approaches indicate significant transfer of dye or melanosomes from melanocytes to keratinocytes that increased with time of co-culturing. Using these model systems, we subsequently tested a battery of lectins and neoglycoproteins for their effect in melanosome transfer. Addition of these selected molecules to co-cultures inhibited transfer of fluorochrome by approximately 15-44% as assessed by flow cytometry, and of melanosomes by 67-93% as assessed by electron microscopy. Therefore, our results suggest the roles of selected lectins and glycoproteins in melanosome transfer to keratinocytes in the skin. PMID- 11434567 TI - Tissue factor expression and serum level in patients with melanoma does not correlate with disease progression. AB - Not only does tissue factor (TF) play a crucial role in hemostasis and thrombosis, but it is also involved in tumor progression and metastatic potency in some malignant tumors. We evaluated the clinical relevance of TF expression in melanocytic tumors and TF serum level in patients with malignant melanoma. TF expression in benign and malignant melanocytic lesions was examined by immunoperoxidase staining in 20 nevi, 41 primary, and 24 metastatic melanoma lesions. TF was detected in 94, 95, and 100% of these lesions, respectively. The staining pattern was membranous and cytoplasmic both in nevi and melanoma cells. This finding was confirmed by western blot analysis using cultured human melanocytes, nevi cells, and melanoma cell lines. TF was also expressed on blood vessels in benign and malignant melanocytic lesions. Expression of TF in primary melanoma lesions was not associated with any clinicopathological variables. In addition, the serum level of TF was elevated in 14% of patients with melanoma; however, it was not correlated with disease progression. These results suggest that TF was ubiquitously expressed in melanocytic cells and its expression was not correlated with disease progression and/or metastatic potency of melanoma cells. PMID- 11434568 TI - UVB-induced decrease of p16/CDKN2A expression in skin cancer patients. AB - The lack of p16 expression has been shown in cultured melanoma cells, however contradictory evidence for p16 expression in melanoma tissues exist. Ultraviolet (UV) C and UVB have been shown to affect p16 expression, which impairs cell cycle regulation in vitro and in vivo. In this study, p16/CDKN2A gene expression was determined by reverse transcription polymerase chain reaction in seven skin cancer patients, in one dysplastic nevus patient and in seven healthy individuals, prior to UVB exposure and at various times after application of one minimal erythema dose (MED). Five of the seven skin cancer patients showed a down regulation of p16/CDKN2A expression after UVB exposure, while controls remained unaltered. The UVB-induced decline of p16/CDKN2A in skin cancer patients might offer new insights into photocarcinogenesis. The putative sequence of events could start with a down-regulation of p16/CDKN2A expression, which would lead to impaired cell cycle regulation. Altered expression patterns of p16/CDKN2A following UVB exposure could be of value for identifying people with an increased risk of UV-induced skin cancer. PMID- 11434569 TI - Over-expression of MSG1 transcriptional co-activator increases melanin in B16 melanoma cells: a possible role for MSG1 in melanogenesis. AB - MSG1 is a 27 kDa nuclear protein that is expressed strongly in melanotic B16 melanoma cells but very weakly in amelanotic B16 cells. Transient expression of B16 cells with an expression vector for MSG1 resulted in an increase in levels of the enzyme dopachrome tautomerase but not tyrosinase, as detected by western blotting. Stable transfection of B16 melanoma cells with plasmids containing the full length MSG1 or its deletion mutants, however, generated cell lines that showed an increase in levels of tyrosinase, dopachrome tautomerase and cellular melanin when compared with control transfected cells. Our results suggest that MSG1 plays an important role in melanogenesis, by regulating the levels of the enzymes of the pigmentary system via tyrosinase and dopachrome tautomerase. PMID- 11434570 TI - Antimalarials--the 'real' advance in lupus. PMID- 11434571 TI - Double trouble: are lupus and malignancy associated? PMID- 11434572 TI - Sticky blood and headache. PMID- 11434573 TI - Systemic lupus erythematosus and the risk of malignancy. AB - The objective of this study was to determine the relative risks of malignancy and of site-specific malignancies in patients with systemic lupus erythematosus (SLE). A cohort of 297 patients (91% Caucasian) with SLE were seen between 1975 and 1994 and followed for a mean of 12 years at the University of Saskatchewan Rheumatic Disease Unit. Expected cancer incidence rates were determined based on Province of Saskatchewan population statistics matched to each study patient for age, sex and calendar year of follow-up. Standardized incidence ratios (SIRs) of observed to expected cancers and 95% confidence intervals (95% CI) were calculated. A total of 27 cases of cancer were observed, whereas only 16.9 were expected (SIR 1.59 (95% CI 1.05-2.32)). For site-specific malignancies, an excess of cancer of the cervix (SIR 8.15 (95% CI 1.63-23.81)) as well as hemopoietic malignancy (SIR 4.9 (95% CI 1.57-11.43)) was found. The hemopoietic cancers were predominantly non-Hodgkin's lymphoma (SIR 7.01 (95% CI 1.88-17.96)). We did not find an association of malignancy with known risk factors, including use of cytotoxic agents. Increased risk of malignancy, notably non-Hodgkin's lymphoma and perhaps cervical cancer, should be regarded as a complication of SLE. PMID- 11434574 TI - Hydroxychloroquine (HCQ) in lupus pregnancy: double-blind and placebo-controlled study. AB - We conducted a randomized, controlled study to assess the need for hydroxychloroquine (HCQ) during lupus pregnancy and to assess safety. Twenty consecutive pregnant patients with similar characteristics were enrolled. The HCQ group included eight patients with systemic lupus erythematosus (SLE) and two with discoid lupus erythematosus (DLE). The placebo (PL) group included nine patients with SLE and one with DLE. The HCQ group had no flare-ups. SLEPDAI scores were similar at study entry, and at conclusion the placebo group had significantly higher scores. One patient had improvement of skin lesions and another of arthritis, allowing a decrease of prednisone dose. There were no retinal effects. Three patients in the PL group flared up, two with skin rashes, one also with arthritis and uveitis, and one (previously in remission on HCQ) with hemolytic anemia, polyserositis and anti-dsDNA antibody. Toxemia was diagnosed in only three patients in the PL group (one fetal death). Comparing prednisone dosage change, we noted a decrease in the HCQ and an increase in the PL group. Delivery age and Apgar scores were higher in the HCQ group. Neonatal examination did not reveal congenital abnormalities, nor did a neuro ophthalmological and auditory evaluation at 1.5-3 y of age. In spite of the small number of patients studied, we noted beneficial effects of HCQ during lupus pregnancy, as measured by SLEPDAI and decrease in prednisone dose with no detriment to patients' health. PMID- 11434575 TI - Reliability and validity of systemic lupus activity measure-revised (SLAM-R) for measuring clinical disease activity in systemic lupus erythematosus. AB - New clinical scales for semiquantitating disease activity in systemic lupus erythematosus (SLE) are widely used in research. They are reliable and valid measures. One of the original scales, the Systemic Lupus Activity Measure (SLAM), has been modified based on experience with it in multi-observer studies and training of individuals in its use. We tested the psychometric properties of the revised SLAM (SLAM-R). SLAM-R was tested on 30 SLE patients, who fulfilled 1997 revised ACR criteria and were selected to represent a range of disease activity. The patients were evaluated independently by two physicians, who studied the instruction booklet and who had never used SLAM-R, on two occasions 2-4 weeks apart. At the first visit, the physician's global assessment of activity using visual analog scale, anti-dsDNA Ab, C3 and C4 were checked for construct validity. The psychometric properties were analyzed with nested analysis of variance and Pearson's correlation coefficient using SAS. All patients were female, the median age was 31 (15-52) y, and the mean score of SLAM-R was 10.5 +/ 5.3 (3-26). Estimates of reliability were 0.78 of inter-rater, 0.61 of inter visit, 0.76 of physician 1 between visits, and 0.56 of physician 2 between visits. Among subcategories except 'Eye,' the 'Gastrointestinal' category had the highest (0.96) and the 'Neuromotor' category had the lowest inter-rater reliability (0.50). With respect to construct validity, the correlation of SLAM-R scores with the disease activity variables except C4 was high and statistically significant. In conclusion, the SLAM-R is reliable and valid for measuring clinical disease activity in SLE. PMID- 11434576 TI - Circulating antibodies to guanosine in systemic lupus erythematosus: correlation with nephritis and polyserositis by acute and longitudinal analyses. AB - Systemic lupus erythematosus (SLE) is characterized by autoantibodies, including antibodies to the nucleosides of DNA. Guanosine is the most immunogenic nucleoside. In this study serum antiguanosine antibody levels were compared with disease activity, determined by their SLEDI score, in 86 patients with SLE. Sera from these patients were tested, by ELISA, for autoantibodies to guanosine, single-stranded DNA (ssDNA), and double-stranded DNA (dsDNA). Anti-double stranded DNA levels were also measured by RIA. Resultant values from these assays were correlated with SLE disease activity, and compared with specific features of SLE. The strongest correlation was higher levels of antiguanosine antibodies in patients with active lupus nephritis and polyserositis compared to patients with inactive disease (P < 0.0001). Antiguanosine levels also correlated with arthritis (P < 0.006), CNS lupus (P < 0.005), and hematologic manifestations of SLE (P < 0.002). To test the validity of this association in chronic SLE, serum antiguanosine antibodies were measured in patients with SLE at various phases of disease activity. Twelve patients with SLE had serum samples drawn at active, active-improved, and inactive phases over a 3-7 y period. Differences were significant for serum antiguanosine antibodies in the active group compared to the inactive group (P < 0.05) and the active vs the active-improved group (P < 0.02), unlike those for dsDNA and ssDNA by ELISA or RIA. Antiguanosine antibodies correlated more closely with disease activity in SLE patients in this longitudinal study than either anti-dsDNA or ssDNA antibodies. Thus, antibodies to guanosine correlated as well or better with disease activity than the other anti-DNA antibodies measured and should be considered to contribute to the pathology of SLE, especially lupus nephritis. PMID- 11434577 TI - Significance of isolated hematuria and isolated pyuria in systemic lupus erythematosus. AB - Hematuria or sterile pyuria as isolated urinary findings present a clinical dilemma for the treating physician. Our objective was to determine whether isolated hematuria and isolated sterile pyuria are associated with active systemic lupus erythematosus (SLE) with respect to renal and non-renal disease activity. This is a descriptive study from a large SLE cohort followed prospectively at the University of Toronto Lupus Clinic. All episodes of isolated hematuria and isolated pyuria between 1970 and 2000 were identified from our database. Isolated hematuria was defined as > 5 red blood cells per high power field; isolated sterile pyuria was defined as > 5 white blood cells per high power field in the absence of urinary infection and other renal manifestations. Non-renal disease activity (defined as nrSLEDAI > 1) was determined at first episode of isolated hematuria and pyuria. Renal disease activity was assessed by scoring renal biopsies within 3 months of detecting isolated hematuria or sterile pyuria. Thirty-four percent (323/946) of our cohort had at least one episode of isolated hematuria. Seventy-seven percent of these patients had concurrent non renal disease activity. Of the 22 biopsies scored with isolated hematuria, 96% were abnormal (WHO > class I), including 52% with active nephritis. Twenty-three percent (215/946) had at least one episode of isolated sterile pyuria. Seventy eight percent of these patients had concurrent non-renal disease activity. All 12 biopsies scored with isolated pyuria were abnormal (WHO Class > 1), including 75% with active nephritis. The appearance of isolated hematuria and isolated pyuria is associated with active renal and non-renal disease activity. An ongoing debate has emerged regarding the significance of isolated hematuria and isolated pyuria with respect to SLE disease activity. The results of this study suggest that isolated hematuria and isolated pyuria is associated with active renal and non renal disease activity. Thus isolated hematuria and isolated sterile pyuria should be considered manifestations of active SLE. PMID- 11434578 TI - Selected serum cytokines in systemic lupus erythematosus treated with quinagolide. AB - The objective of this study was to determine the effect of quinagolide (Norprolac) on serum level of cytokines in systemic lupus erythematosus (SLE) patients. In 20 SLE patients treated with a low dose of quinagolide, and in 17 healthy persons who constituted the control group, concentration of serum prolactin (PRL), interleukins (ILs), soluble tumor necrosis factor receptors (sTNF Rs) preceded by calculation of disease activity index (SLEDAI) were tested at entry and then after 3 months in 16 patients and after 6 months in 11 patients who completed the study. Serum PRL level was higher (though insignificantly) in the SLE group than in the controls and decreased significantly after 6 months of therapy. A raised SLEDAI score at entry was significantly reduced during therapy but a weak correlation with PRL level was revealed. A significant increase in IL 6 level in SLE group as compared to controls was observed (respectively 14.57 +/- 13.25 and 5.04 +/- 3.35 microg/ml) as well as a significantly decreased level after 6 months of treatment (4.30 +/- 2.51 pg/ml). There was a significant difference between sTNF RI concentration before and after 3 months of quinagolide treatment (respectively 1140.83 +/- 312.08 and 1454.58 +/- 465.54 pg/ml). After 6 months of treatment a statistically significant correlation between concentration of PRL and level of IL-6 and a negative correlation between PRL and sTNF RI was revealed. Quinagolide treatment may have a role in the management of SLE patients. PMID- 11434579 TI - Osteoporosis in murine systemic lupus erythematosus--a laboratory model. AB - The aim of this study was to assess the skeletal metabolism in a murine model of systemic lupus erythematosus (SLE). MRL/n and MRL/l mice (respectively representing a benign and a malignant form of the disease) were observed from 1.5 to 6.5 months of life. The monthly follow-up included: biochemical and histomorphometrical studies of the femoral bone, serum biochemistry, immunoglobulins and osteocalcin, and histological evaluation of the kidney tissue. The results showed a higher femoral weight (+11.5%), calcium (+4.4%) and protein bone content (+11.4%) and a significantly higher (+77%) phosphorus bone content in the MRL/n group; significantly lower (-48.9%) bone alkaline phosphatase enzymatic activity, lower bone alkaline/acid phosphatase enzymatic activities ratio (-40.8%) and lower (-38.4%) serum osteocalcin values in the MRL/l group (which might suggest reduced bone formation in these animals); markedly smaller trabecular bone volume (BV/TV) in the femoral head (-36.2%) and femoral neck (-39.8%), and smaller cortical and femoral areas in the mid-femoral shaft (-38.8% and -38.1% respectively) in the MRL/l group; higher serum immunoglobulins, increased serum blood urea nitrogen (BUN) and creatinine and a higher index of activity in the kidney histology in the MRL/l group, indicating increased activity of the disease in this substrain. The MRL mice, through their two substrains, may serve as a valuable laboratory animal model for study of the skeletal changes in SLE and of the influence of the disease activity on the skeletal metabolism. PMID- 11434580 TI - Systemic lupus erythematosus in Aborigines and Caucasians in central Australia: a comparative study. AB - The objective of this study was to determine whether there are differences in the prevalence, clinical and laboratory manifestations, and morbidity and mortality of systemic lupus erythematosus (SLE) between Aborigines and Caucasians in Central Australia. The medical records of all patients diagnosed with SLE upto December 1999 were reviewed retrospectively. Prevalence of SLE was 1:1360 for Aborigines and 1:5170 for Caucasians. The prevalences of malar rash, discoid rash, photosensitivity, oral ulcers, pleuritis, anticardiolipin antibodies and lupus anticoagulant were higher in Caucasians than in Aborigines. The prevalences of anti-Sm antibody and anti-RNP antibody were higher in Aborigines than in Caucasians. These differences did not attain statistical significance. There was a low prevalence of renal disease in Aborigines and Caucasians. Mortality was low in Aborigines and nil in Caucasians. Although there is a high prevalence of SLE in Aborigines in Central Australia, renal involvement and mortality are low. PMID- 11434581 TI - Systemic lupus erythematosus with polyglandular autoimmune syndrome type II: report of an unusual case. AB - A patient with systemic lupus erythematosus (SLE) is described who had associated autoimmune thyroiditis with subclinical hypothyroidism and idiopathic Addison's disease. The presence of autoimmune thyroid disease and Addison's disease could classify her as having polyglandular autoimmune syndrome (PGAS) type II, which is the commonest of the three types of this syndrome. However, the additional presence of SLE in PGAS type II has not been described earlier. This patient appears to be the first such case. PMID- 11434582 TI - Lupus flare associated with growth hormone. AB - This case illustrates a young man with an 11 y history of systemic lupus erythematosus who developed lupus flare after a 9-month use of growth hormone (GH) for the treatment of growth retardation. The course of disease was improving after GH cessation and treatment with high dose corticosteroids and intravenous cyclophosphamide pulse therapy. PMID- 11434583 TI - Risk factors related to hypertension in a Spanish systemic lupus erythematosus cohort. PMID- 11434584 TI - Nonsteroidal anti-inflammatory drugs, low-dose aspirin, and potential ways of reducing the risk of complications. AB - Use of nonsteroidal anti-inflammatory drugs (NSAIDs) is associated with an annual incidence of 1-2% of gastrointestinal (GI) complications, which may be fatal in some cases. Low-dose aspirin is also associated with an increased risk of upper GI bleeding, but the increase is about three times lower than that found with common NSAIDs. Misoprostol (600-800 microg/day) reduces the incidence of complications in non-aspirin NSAID users. Co-therapy with antisecretory drugs (proton pump inhibitors, PPIs) also reduces the risk of GI bleeding in high-risk patients taking NSAIDs and/or low-dose aspirin. Another way of reducing the incidence of GI complications is to use the highly selective cyclo-oxygenase 2 (COX-2) inhibitors. The GI safety of nitric oxide NSAIDs (NO-NSAIDs) has been demonstrated extensively in experimental conditions and preliminary clinical studies. Epidemiological studies have also shown that nitric oxide donor drugs reduce the risk of upper GI bleeding, which might be important in patients receiving low-dose aspirin. PMID- 11434585 TI - Barrett's oesophagus: better left alone? AB - The risk of oesophageal adenocarcinoma in patients with Barrett's oesophagus is overrated in most publications, including meta-analyses. The upper limit of the overall risk is currently estimated at 0.5 per 100 patients per year. This means that one cancer may occur out of 200 patients followed for one year. Furthermore, a large fraction of patients with Barrett's oesophagus have a precarious health status, either from advanced age or from a chronic severe disease. Management is based upon a careful index endoscopy; accordingly, patients positive for dysplasia must be strictly observed or treated. Endoscopic surveillance protocols aiming at early detection of neoplasia--and treatment--in patients without dysplasia are unjustified in most cases. On the other hand, a minority of male patients--providing a good performance status--should enter into surveillance protocols when high-risk factors such as a long history of reflux symptoms and smoking habits are present. PMID- 11434586 TI - Of tubes and men: studying manometrically the effects of laxatives on colonic motility. AB - Chronic constipation is encountered frequently in clinical practice, and the use of laxatives is widespread in the general population. The need for effective and safe compounds stimulates research in the field. Unfortunately, to assess the colonic motility effects of these compounds, we have to rely on objective measurements available only through relatively invasive studies, such as manometric recordings of the motor activity of the large bowel. However, the possibility of studying almost the entire length of the viscus with these techniques allows a better definition of the pathophysiological processes involved in the genesis of constipation, and more insights into the effects of specifically acting drugs. PMID- 11434587 TI - IgA anti-tissue transglutaminase: setting the stage for coeliac disease screening. AB - Coeliac disease is triggered in genetically predisposed individuals by the ingestion of wheat and related cereals. Affected persons raise an intestinal mucosal T-cell response against the gluten fraction of these cereals. Furthermore, they produce characteristic circulating IgA antibodies to a self antigen present in the extracellular matrix that can be detected on tissue sections. The positive predictive value of these endomysial, reticular or umbilical cord antibodies for coeliac disease comes close to 100%. Recently, the enzyme tissue transglutaminase was identified as the main, if not sole, endomysial autoantigen in coeliac disease. Enzyme-linked immunosorbent assay tests with tissue transglutaminase from guinea pig or the recombinant human enzyme have been established that allow a standardized and quantitative determination of IgA anti-tissue transglutaminase titres. While the published assay variants report high positive and negative predictive values for coeliac disease, they were applied to preselected patients from mostly single centres. Therefore, validation and in part cross-validation of a standardized assay based on guinea pig tissue transglutaminase in 38 European and non-European centres is timely. With a sensitivity of 90% and specificity of 96% relative to local diagnostic standards the assay performed well. Considering further improvement by the use of recombinant human tissue transglutaminase as the antigen and central re-evaluation of the local standards for confirmation of coeliac disease, this enzyme-linked immunosorbent assay promises to become the primary tool for non invasive diagnosis, therapy control and screening of coeliac disease. However, with an estimated prevalence of 1:100-1:200 of mostly atypical and subclinical coeliacs in Western populations, we are confronted with the question of how far mass screening is ethically feasible and cost effective. PMID- 11434588 TI - A majority of patients with Barrett's oesophagus are unlikely to benefit from endoscopic cancer surveillance. AB - BACKGROUND: Endoscopic cancer surveillance has been advocated for patients with Barrett's oesophagus. However, only a small minority of patients dies from adenocarcinoma in Barrett's oesophagus. It has been calculated that endoscopic cancer surveillance will only add to the quality of life of individuals in whom the incidence of adenocarcinoma in Barrett's oesophagus is greater than 1/200 patient-years. OBJECTIVE: To determine the proportion of a consecutive cohort of patients, in whom Barrett's oesophagus was diagnosed over a 5-year period, likely to benefit from endoscopic cancer surveillance. METHODS: All patients who had died during the observation period or were over 75 years old and those with diseases likely to impair survival were excluded. Next, all patients in whom the risk of developing adenocarcinoma in Barrett's oesophagus fell below 1/200 patient-years were excluded (including all women, all men under the age of 60 and all men with Barrett's oesophagus of < 3 cm in length). Patients with dysplasia of any degree and/or presence of an ulcer or stricture in Barrett's oesophagus were reinstated. RESULTS: Of 335 adult patients diagnosed with Barrett's oesophagus but without adenocarcinoma or high-grade dysplasia, 75 had died from unrelated causes, 47 had other diseases limiting survival and 59 were over 75 years old. After exclusion of all women, all men with Barrett's oesophagus of < 3 cm in length and all men under 60 years old, 15 patients were left. However, 32 were reinstated because of risk factors and another five because of insufficient data, resulting in 52 of the original 335 patients (15.5%) being eligible for endoscopic cancer surveillance. CONCLUSION: This study suggests that less than 20% of patients identified with Barrett's oesophagus at routine endoscopy would benefit from endoscopic cancer surveillance. Prospective surveillance programmes should be limited to patients with an increased cancer risk and a good health profile. PMID- 11434589 TI - Effects of polyethylene glycol 4000 on 24-h manometric recordings of left colonic motor activity. AB - BACKGROUND: It has been shown that low doses of polyethylene glycol (PEG) 4000 are effective in the treatment of chronic constipation. The aim of this study was to describe the effects on colonic motility of oral PEG 4000 treatment and intraluminal instillation of PEG 4000. METHODS: Left colonic and rectosigmoid manometric recordings were performed for 27 h in six constipated patients and in six healthy volunteers. At the end of the recording, bisacodyl and PEG 4000 were instilled into the lumen of the colon. To assess the effects of oral administration of PEG 4000 on colonic motility, manometric recordings were also performed in constipated patients after 4 weeks of treatment with PEG 4000. RESULTS: All patients had significantly more stools during than before PEG treatment. There was no significant difference between the number and the characteristics of high-amplitude propagating contractions (HAPC) or the area under the curve (AUC) before or during treatment with PEG 4000. Intraluminal instillation of PEG induced HAPC in only one patient and in no controls. CONCLUSION: This study shows that PEG 4000 has no effect on left colonic and rectosigmoid motor activity during oral treatment, despite its clinical effectiveness, or after local instillation. PMID- 11434590 TI - Open-access versus hospital-initiated flexible sigmoidoscopy: a comparative audit of efficacy. AB - BACKGROUND AND AIMS: There is a continuing demand for the provision of open access endoscopy services. While open-access gastroscopy is now a well established practice in most hospitals, open-access flexible sigmoidoscopy (OAS) is not yet available to the same extent. We present our early experience with OAS, a new service provided by our unit. PATIENTS, METHODS AND RESULTS: Over a 12 month period, 255 flexible sigmoidoscopies were carried out (139 OAS and 116 hospital-initiated sigmoidoscopies, HIS). Referral criteria for sigmoidoscopy were similar in both groups, with rectal bleeding and change in bowel habit being the most frequent. RESULTS: The two groups were comparable with regards to age and gender, but patients referred for OAS had symptoms for shorter duration than those referred for HIS (30 +/- 5.6 months v. 101 +/- 28 months, P < 0.00001). The diagnostic yield for OAS was 44% and for HIS 29% (P= 0.01). Significant pathology (colorectal cancer, polyps and newly diagnosed inflammatory bowel disease, colonic strictures) was detected in 31 patients (22%) in the OAS group and 12 patients (10%) in the HIS group (P = 0.02). Six cancers were diagnosed (5 OAS, 1 HIS), all staged Duke A. The waiting times for OAS and HIS were 3.1 +/- 0.3 weeks and 11.4 +/- 1.2 weeks, respectively (P = 0.0001). The diagnostic yield of fibre optic endoscopy was independent of duration of symptoms, although patients referred for OAS with rectal bleeding and/or diarrhoea had a higher diagnostic yield (55/125, 44%). CONCLUSION: OAS is feasible and effective, with a high diagnostic yield. Such service can usually be provided with minimal added resources. PMID- 11434591 TI - The serum IgA class anti-tissue transglutaminase antibodies in the diagnosis and follow up of coeliac disease. Results of an international multi-centre study. International Working Group on Eu-tTG. AB - OBJECTIVES: So far the reliability of the anti-tissue transglutaminase (anti-tTG) test for the diagnosis of coeliac disease has mostly been evaluated using slightly different enzyme-linked immunosorbent assays (ELISAs) in selected and usually small groups of patients. The aims of this study were: (1) to evaluate the reliability of the IgA anti-tTG antibodies for the diagnosis of coeliac disease; and (2) to define the sensitivity and specificity of a commercially available kit for the anti-tTG antibodies' quantitative determination. DESIGN: Each centre in this international multi-centre study collected sera from three groups of subjects: coeliac disease patients at the onset of (1) or on a gluten free diet for at least 12 months (2); disease and healthy controls (3). METHODS: The anti-tTG antibodies were determined in duplicate using an ELISA-based commercially available kit (Eu-tTG Eurospital, Trieste, Italy). RESULTS: The following overall cases and controls have been enrolled: (1) 399 subjects with active coeliac disease; (2) 351 treated coeliac disease cases; (3) 432 controls. The centralized re-testing was performed on: (1) group a: 176 patients with active coeliac disease (mean anti-tTG, 21 arbitrary units [AU]); (2) group b: 172 treated coeliac disease cases (mean anti-tTG, 5 AU); (3) group c: 206 controls (mean anti-tTG, 3 AU). In active coeliacs, the anti-tTG antibodies showed a significant progressive decrease with age, while in controls an opposite trend was found. In active coeliac disease patients, the anti-tTG antibodies were significantly higher in coeliacs with a grade III enteropathy than in those showing a grade II lesion. In treated coeliacs, the mean anti-tTG values were significantly lower in patients strictly adhering to a gluten-free diet than in those reporting dietary transgressions. The sensitivity and the specificity of the Eu-tTG assay were 90% and 96%, respectively. CONCLUSION: The results of this study show that the commercially available test for the anti-tTG antibodies' determination is a reproducible and valuable tool for the diagnosis and follow up of coeliac disease. PMID- 11434592 TI - Adult endomysial antibody-negative coeliac disease and cigarette smoking. AB - OBJECTIVE: To determine the relative incidence and characteristics of endomysial antibody (EMA)-negative coeliac disease in adults. DESIGN: Retrospective analysis of prospectively collected data on adults with newly diagnosed coeliac disease, with determination of EMA status before gluten withdrawal. SETTING: District general hospital (secondary care institution). PARTICIPANTS: Sixty consecutive incident cases. MAIN OUTCOME MEASURES: (i) Proportion of cases who were EMA negative; (ii) comparison of clinical and laboratory variables at diagnosis for EMA-positive and EMA-negative subjects. RESULTS: Fifteen subjects (25%, 95% CI 15 38%) were EMA negative, of whom only two were IgA deficient. There was clinical evidence in all 15 patients and histological evidence in 13 patients of a response to gluten withdrawal. No significant differences were found between EMA positive and EMA-negative subjects with respect to histological features, age, gender, clinical manifestations, concurrent autoimmune disorders, family history of coeliac disease, or haemoglobin and albumin concentrations at diagnosis. However, EMA-negative status at diagnosis was associated strongly with current or recent cigarette smoking (OR 7.0, 95% CI 1.7-31.5, P= 0.003). CONCLUSIONS: A substantial minority of patients with otherwise typical coeliac disease are EMA negative, and most of these are IgA replete. The value of EMA as a screening tool is therefore limited. EMA status in untreated coeliac disease correlates strongly with cigarette smoking history: this may be of pathogenic significance, given the previously demonstrated association between smoking and the risk of coeliac disease. PMID- 11434593 TI - Validity of a Helicobacter pylori stool antigen assay for the assessment of H. pylori status following eradication therapy. AB - BACKGROUND: A Helicobacter pylori stool antigen (HpSA) test has been proposed as a valid alternative to the 13C-urea breath test (13C-UBT) for the non-invasive detection of H. pylori infection in primary diagnosis. Published reports show conflicting results with regard to the test's diagnostic accuracy after eradication therapy. The aim of the present study was to assess the diagnostic value of the HpSA test and to determine the optimal discriminating cut-off value in patients following H. pylori eradication therapy. METHOD: Stool samples were collected and the 13C-UBT was performed in 113 patients 4-6 weeks after eradication therapy. A validated test protocol for the 13C-UBT was used. Stool specimens were analysed with the Premier Platinum HpSA enzyme immunoassay (EIA). A receiver operator characteristics (ROC) analysis was performed to define the optimal cut-off value on the basis of the results of the 13C-UBT. RESULTS: The results of the 13C-UBT showed that H. pylori eradication was successful in 83/113 (73%) patients. According to the manufacturer, the cut-off value for the HpSA test is 0.14 optical density, but this does not appear to be valid after eradication therapy (sensitivity 76.7%, specificity 98.8%). On the basis of ROC analysis, the optimal cut-off value after therapy was determined to be 0.11 optical density, giving a sensitivity of 93.3% and a specificity of 93.9%. CONCLUSION: The HpSA test is a valid test for the assessment of H. pylori status after eradication therapy, provided an adjusted cut-off value is applied. PMID- 11434594 TI - Helicobacter pylori infection in patients consulting gastroenterologists in France: prevalence is linked to gender and region of residence. AB - BACKGROUND: Because of limited data on the epidemiology of Helicobacter pylori in France, the prevalence of this infection by region and its associated risk factors were studied between 1995 and 1997 among patients consulting a representative sample of gastroenterologists by region. METHOD: A cross-sectional study was performed. Patients consulting gastroenterologists for whatever reason were screened for H. pylori infection determined by specific salivary IgG. A questionnaire was filled out by the gastroenterologist. A multivariate analysis was performed with all relevant variables. RESULTS: 3,153 patients were included. The mean age was 48.5 years; 51.8% were women. After stratification by patients consulting for upper digestive tract (UDT) and non-UDT symptoms, H. pylori infection was found to be more prevalent, in both groups, for characteristics such as being born in a developing country, overcrowding during childhood, and primary educational level. Interestingly, gender (odds ratio OR(UDT for women) = 0.7 (95% CI 0.5-0.8] and OR(non-UDT) for women = 0.6 [95% CI 0.5-0.8]) and living in a region other than the south-west (OR(UDT) varying from 1.5 to 2.0 and OR(non UDT) varying from 1.3 to 2.1, depending on the region) was associated with the odds of prevalent infection. CONCLUSION: These findings show (1) that gender deserves more attention in the epidemiology of H. pylori and (2) a regional disparity in France regarding H. pylori infection. PMID- 11434595 TI - The AMOR study: a randomized, double-blinded trial of omeprazole versus ranitidine together with amoxycillin and metronidazole for eradication of Helicobacter pylori. AB - BACKGROUND: Besides antibiotics, additionally effective acid inhibition is necessary for the eradication of Helicobacter pylori. OBJECTIVE: To assess the significance of acid suppression and, in particular, treatment with proton pump inhibitors (PPIs) compared with H2 receptor antagonists (H2 RAs). The primary target parameter for the study was H. pylori eradication. In addition, the ulcer healing rate, speed of pain reduction, score for gastritis in the antrum and gastric body, and rate of side effects were recorded. DESIGN: Randomized, double blinded, multicentre study. PARTICIPANTS: A total of 456 patients between the ages of 18 and 80 years with H. pylori-positive duodenal ulcers were included in the study. METHODS: Using a randomization list, patients were assigned either to a treatment group receiving omeprazole 40 mg once daily, amoxycillin 750 mg three times a day, and metronidazole 500 mg three times a day (OAM), or to a group receiving ranitidine 300 mg once daily, amoxycillin 750 mg three times a day, and metronidazole 500 mg three times a day (RAM). The treatment period was 7 days in both groups. Long-term acid-suppressant treatment was not given. RESULTS: The eradication rate was 87.1% (169/194, intention to treat [ITT]) in the OAM group and 77% (137/ 178, ITT) in the RAM group. The difference of 10.1% (95% CI 2.5 18%) is statistically significant (P= 0.0104). The ulcer healing rate was 93.3% in the OAM group (181/194, ITT) and 92.1% in the RAM group (164/178, ITT, NS). With regard to the speed and intensity of pain reduction, the OAM group was superior to the RAM group. In patients in whom H. pylori eradication was successful, the reduction in the antral and gastric body gastritis score was significantly greater than in patients without eradication. In the OAM group, 39.1% of the patients (n = 90) reported one or more side effects, compared with 44.7% (n = 101) in the RAM group (P= 1.5449, NS). CONCLUSION: Omeprazole (40 mg once daily in the morning) is significantly more effective than ranitidine (300 mg once daily in the morning) with respect to H. pylori eradication when used together with amoxycillin (750 mg three times a day) and metronidazole (500 mg three times a day) for a 7-day treatment period. PMID- 11434596 TI - Metabolic heterogeneity of glycogen in hepatocytes of patients with liver cirrhosis: the glycogen of the liver lobule zones in cirrhosis. AB - The concentrations of total glycogen (TG) and its labile (LF) and stable (SF) fractions were determined in hepatocytes of portal and central zones of the normal human liver and in the liver of patients with cirrhosis of viral and alcohol aetiologies. Using PAS reaction, TG, LF and SF were revealed in histological sections of the material obtained by the liver punch biopsies. The concentrations of TG and its fractions were measured by televisional cytophotometry. In liver cirrhosis, the concentrations of TG, LF and SF in both zones of the hepatic lobule have been found to be much higher than in the normal liver. It has been shown that the ratio of the hepatocyte TG concentrations in the portal zone to the central zone both in the normal liver and in viral cirrhosis exceeds 1.0, amounting to 1.264 +/- 0.021 and 1.030 +/- 0.009, respectively. The glycogen fraction composition in the cells of both the liver lobule zones in viral cirrhosis does not differ significantly from the norm. On the contrary, in the liver of patients with alcoholic cirrhosis, the ratio of the TG concentrations in the portal zone to the central zone is reduced to 0.815 +/- 0.016 and is accompanied by qualitative changes of the glycogen composition. PMID- 11434597 TI - Treatment of patients with chronic hepatitis C not responding to interferon with high-dose interferon alpha with or without ribavirin: final results of a prospective randomized trial. Austrian Hepatitis Study Group. AB - OBJECTIVE: To investigate the efficacy of high-dose interferon alpha (IFN-alpha) with or without ribavirin in interferon (IFN) non-responders. STUDY DESIGN: 304 chronic hepatitis C patients received 5 MU IFN-alpha2b (IntronA, Schering-Plough, Kenilworth, NJ, USA) three times a week for 3 months. Non-responders were randomized either to continue with IFN (IFN 5 MU/TIW followed by 10 MU/TIW, each for 3 months) alone (group A: n = 76, m: f = 54: 22, age 45.7 +/- 12 years, 16% cirrhosis, alanine aminotransferase [ALT] 66 +/- 35 U/l) or in combination with ribavirin (approximately 14 mg/kg/day) (group B: n = 81, m: f = 57: 24, age 48.2 +/- 12 years, 17% cirrhosis, ALT 71 +/- 40 U/l). At the end of treatment, patients were followed for 6 months. MAIN OUTCOME MEASURES: Virological response at end of treatment and 6 months thereafter. SETTING: University hospitals and tertiary referral centres. RESULTS: At the end of treatment, eight (10.8%) and 25 (31.3%, P= 0.0066) patients were HCV-RNA negative, and 51 and 39 were HCV-RNA positive, in groups A and B, respectively. There were 17 drop-outs in each group. Six months after treatment, only one patient in group A (1.3%) and seven patients (8.6%, P= 0.06) in group B had normal ALT and undetectable serum HCV-RNA. CONCLUSIONS: A combination of high-dose IFN with ribavirin induces a short lasting complete response in about one-third of IFN-non-responders. PMID- 11434598 TI - Ischaemic jejunal stenosis complicating portal and mesenteric vein thrombosis: a report of two cases. AB - A major complication of portal and mesenteric vein thrombosis is acute bowel ischaemia resulting in infarction and requiring immediate resection of the involved segment. Sufficient collaterals can prevent acute haemorrhagic infarction, but bowel stenosis due to chronic ischaemia may develop. We report two cases of ischaemic jejunal stenosis occurring 4 weeks after successful treatment of portal and mesenteric vein thrombosis. Diagnosis of high-grade segmental stenosis of the jejunum was established by contrast medium radiography of the gastrointestinal tract. After laparotomy and resection of the stenosed jejunal segment, both patients recovered well from the operation and were released from hospital. Follow-up examinations revealed an unremarkable state of health. Ischaemic bowel stenosis should be considered in patients with recurring abdominal pain after mesenteric and portal vein thrombosis. A close follow-up of every patient after treatment for mesenteric and portal vein thrombosis should be carried out to ensure early diagnosis of this complication. PMID- 11434599 TI - A new mutation of E-cadherin gene in familial gastric linitis plastica cancer with extra-digestive dissemination. AB - Over a 12-month period, we diagnosed poorly differentiated infiltrative independent-cell gastric adenocarcinoma in two brothers and one sister aged 41 to 47 years. Their father had died from antral cancer at the age of 34 years. These cancers had two characteristic clinical features: rapid course and distant malignant dissemination. In all three patients, polymerase chain reaction sequencing of the E-cadherin (CDH1) gene of white blood cells identified a heterozygous nonsense mutation of exon 3, producing a stop codon at position 95 (Q95X), resulting in a truncated protein. The alteration of this protein, which plays a crucial role in epithelial cell adhesion, probably explains the clinical expression in this type of familial diffuse gastric cancer. PMID- 11434600 TI - Angiodysplasia in a duodenal diverticulum as an unusual cause of upper gastrointestinal bleeding. AB - The majority of duodenal diverticula are asymptomatic but may also induce major haemorrhage on rare occasions. Gastrointestinal bleeding due to angiodysplasia in a duodenal diverticulum is very rare. We present a 70-year-old woman with repeated melaena in whom the diagnosis of angiodysplasia in a diverticulum of the fourth part of the duodenum could be made by standard upper endoscopy. PMID- 11434601 TI - Synchronous first manifestation of an idiopathic eosinophilic gastroenteritis and bronchial asthma. AB - Eosinophilic gastroenteritis is a rare disease of the gastrointestinal tract in which the eosinophils seem to play an important role in the inflammation of the gut wall. We report on a case with a synchronous first manifestation of eosinophilic gastroenteritis and bronchial asthma, which also occurred synchronously in all further episodes. The diagnosis was first made at the end of the second episode during which the patient lost more than 13 kg in weight. Under steroid therapy, symptoms of both diseases disappeared quickly in the third episode. We assume that participation of the gastrointestinal tract in patients with bronchial asthma occurs more frequently than expected. In asthma patients with abdominal symptomatology, eosinophilic gastroenteritis should also be considered. PMID- 11434602 TI - Pancreatitis and haemobilia due to arterioportal fistula after percutaneous liver biopsy resolved by selective arterial embolization. AB - Haemobilia and arterioportal fistula are uncommon complications secondary to percutaneous liver biopsy. We report the case of a patient who developed haemobilia and subsequently acute pancreatitis as a result of a liver biopsy. Selective hepatic angiogram showed an arterioportal fistula. Transcatheter arterial embolization successfully occluded the fistula. The patient remained asymptomatic 4 months later. We review the published literature concerning acute pancreatitis associated with haemobilia and draw conclusions for management of similar cases in the future. PMID- 11434603 TI - Angiosarcoma variant of Kasabach-Merritt syndrome. AB - Kasabach-Merritt syndrome is characterized by the occurrence of disseminated intravascular coagulation (DIC) usually caused by benign angiomatous tumours. Here we report the case of a 70-year-old man in whom DIC revealed a locally advanced hepatic tumour. Although DIC resolved with heparin, antithrombin III, fresh frozen plasma and corticosteroids, the patient died from haemoperitoneum following a fall, 3 months after the initial observation. Histopathological examination by autopsy allowed the diagnosis of hepatic angiosarcoma. The physiopathogenic mechanisms and treatment options are discussed. PMID- 11434604 TI - Candida colitis in a middle-aged male receiving permanent haemodialysis. AB - Candida infections of the gastrointestinal tract are not rare, especially in immunocompromised patients. We present a case of candida colitis in a patient receiving permanent haemodialysis. Candida colitis can be a life-threatening condition and should be treated promptly, especially in immunocompromised patients. PMID- 11434605 TI - Ornidazole-induced autoimmune hepatitis. AB - Ornidazole is a commonly prescribed antiparasitic drug for parasitic infestations, including amoebiasis, giardiasis and Trichomonas vaginalis. Several cases of antibiotic-induced autoimmune hepatitis (AIH) or AIH-like syndrome have been reported recently. In this report, we describe a 35-year-old woman with two relapses of AIH induced by ornidazole prescribed for diarrhoea and vaginal infection. PMID- 11434606 TI - Analysis and clinical effects of gluten in coeliac disease. AB - The prolamin working group coordinates research on laboratory gluten analysis in food and on clinical evaluation of patient sensitivity to prolamins. As an observer organization to the Codex Alimentarius Commission, the group summarizes current data on analysis and effects of gluten in coeliac disease. All types of gliadin, the ethanol-soluble fraction of gluten, contain the coeliac-active factor. However, coeliac toxicity and immunogenicity (humoral and cellular) of various prolamins are not identical in coeliac patients. There are no conclusive data on the threshold of gluten sensitivity of coeliac patients. Information as to the long-term risk to coeliac patients exposed to small doses of gliadin is lacking. Therefore, every effort should be made to keep the diet of coeliac patients as gluten-free as possible. The prolamin group is currently evaluating a new enzyme-linked immunosorbent assay (ELISA) protocol for gluten analysis that could serve as a basis for further Codex regulations. The group recommends adherence to a single Codex limit for gluten-free foods. The current limit of 200 ppm gluten is questionable and requires reconsideration based on new information that will be available soon. PMID- 11434607 TI - Incidence of chronic pancreatitis in the Czech Republic. AB - There have been only a few studies dealing with the incidence of chronic pancreatitis published. Over the last 80 years, original studies describing the incidence of chronic pancreatitis were undertaken, including studies in Scandinavia, Switzerland, Hungary and Poland. Incidence varied geographically, from 1.6 new cases per year among 100,000 inhabitants in Switzerland to 23 new cases among 100,000 inhabitants in Finland. The aetiology of 70% of all cases was alcoholic pancreatitis. The incidence of chronic pancreatitis in the Czech Republic is 7.9 per 100,000 inhabitants per year, similar to that of Denmark and Germany, but higher than that of Poland or Switzerland. In our study, the main aetiological factor was alcohol (65.4% of all cases, with consumption of alcohol of > 75.0 g/day). PMID- 11434608 TI - Long-term management of gastro-eosophageal reflux disease with omeprazole or open antireflux surgery. PMID- 11434609 TI - In vivo angiogenesis in normal and portal hypertensive rats: role of basic fibroblast growth factor and nitric oxide. AB - BACKGROUND: Angiogenesis plays a pivotal role in many processes. Here, we studied whether angiogenesis to basic fibroblast growth factor (bFGF) in normal and portal hypertensive rats requires nitric oxide (NO). METHODS: To measure angiogenesis in vivo, two Teflon rings filled with collagen I (Vitrogen 100) were fixed in the mesenteric cavity at day 0, with one supplemented with bFGF (100 ng). Portal hypertension was induced by partial portal vein ligation (PVL). Sham operated rats served as controls (CON). The role of NO was tested by adding the NO formation antagonist N(omega)-nitro-L-arginine (NNA; 3.3 mg/kg per day) to the drinking water. After 16 days, rings were explanted and embedded, and vessels were morphometrically counted. RESULTS: bFGF significantly stimulated vessel formation per implant in CON rats (from 624 +/- 97 without stimulation to 1123 +/ 171, n = 11, P < 0.01), but not in PVL rats (from 1106 +/- 174 without stimulation to 1046 +/- 202, n = 9). Without stimulation, numbers of ingrown vessels were significantly (P < 0.05) higher in PVL compared to CON rats. NNA substantially inhibited angiogenesis in both groups (P < 0.01). Vessel numbers were 202 +/- 124 for PVL (n = 5) and 197 +/- 14 for CON (n = 5) animals. bFGF did not reverse angiogenesis prevented by NNA (373 +/- 98 for PVL, 265 +/- 26 for CON, n = 5 per group, NS). CONCLUSIONS: NO formation inhibition diminishes both unstimulated and bFGF-stimulated angiogenesis in CON rats. Moreover, bFGF cannot rescue NNA-inhibited angiogenesis in PVL rats. PMID- 11434610 TI - Hepatopulmonary syndrome in candidates for liver transplantation. AB - BACKGROUND: Hepatopulmonary syndrome (HPS) has been defined as a clinical triad, including chronic liver disease, gas exchange defects (increased alveolar arterial PO2 difference irrespective of the presence of arterial hypoxemia), and widespread intrapulmonary vascular dilatations. We determined the incidence and the clinical and pulmonary functional characteristics of HPS in candidates for orthotopic liver transplantation (OLT) and tested their predicted accuracy. METHODS: We studied 80 patients with cirrhosis prospectively, and carried out contrast-enhanced (CE) echocardiography and lung function tests, including ventilation-perfusion (V(A)/Q) distributions. RESULTS: Fourteen patients had HPS (incidence, 17.5%). Patients with HPS (49 +/- 12 (+/-SD) years) had more cutaneous spiders, finger clubbing and dyspnea (P < 0.05 each) and a lower diffusing capacity (DLCO, 56 +/- 18% predicted; P < 0.001) than non-HPS patients (n = 66). Mild to moderate V(A)/Q inequalities and increased intrapulmonary shunt were predominant in HPS patients, but oxygen diffusion impairment was observed in those with hypoxemia (n = 8) only. The DLCO showed a considerable area under the receiver operating characteristic curve (0.89). CONCLUSIONS: HPS in cirrhotic patient candidates for OLT shows a high incidence and these patients present with distinctive clinical and functional features compared with non-HPS individuals. The presence of a low DLCO may be of help for the diagnosis of HPS. PMID- 11434611 TI - Amino acid challenge in patients with cirrhosis: a model for the assessment of treatments for hepatic encephalopathy. AB - BACKGROUND/AIMS: To mimic episodic hepatic encephalopathy after gastrointestinal bleeding under controlled conditions, cirrhotic patients were challenged with an amino acid mixture of comparable composition to haemoglobin. METHODS: Basal EEG, psychometric score (HE test), reaction times and venous blood ammonia were recorded. Following a 54 or 108 gm oral amino acid challenge, blood ammonia levels and EEG were recorded at 30-min intervals, and psychometric testing was repeated at 180 min. Ten controls (57 +/- 2) and 31 cirrhotics (52 +/- 2) of which 21 were Child's grade A or B and 10 grade C underwent the challenge. Nine had a transjugular intrahepatic porta-systemic shunt in situ. RESULTS: Seventeen patients had abnormal baseline HE scores. Basal blood ammonia and reaction time A were significantly greater in patients (52 +/- 5 micromol/l and 478 +/- 20 ms, respectively) than controls (19 +/- 2 micromol/l and 372 +/- 14 ms) (P < 0.001). Following the challenge, in patients with advanced liver disease (Child's grade B and C) the slowing of reaction time A (+85 +/- 38 and +71 +/- 31 ms, respectively; P < 0.03) and EEG (ratio of slow to fast wave activity +0.31 +/- 0.12 and +0.58 +/- 0.19; P < 0.02) were significantly greater than in controls ( 3.3 +/- 8 ms and 0.00 +/- 0.03, respectively). Patients with an abnormal basal HE score had the most pronounced changes (reaction time A +110 +/- 39 ms, P < 0.01, EEG +0.52 +/- 13, P < 0.01, respectively). The change in EEG ratio correlated with the dose of amino acid administered (r = 0.96; P < 0.008). CONCLUSION: The amino acid challenge constitutes a reproducible human model of episodic, Type C hepatic encephalopathy unaffected by the complications usually encountered in clinical practice. PMID- 11434612 TI - Sequence of events in the assembly of Mallory body components in mouse liver: clues to the pathogenesis and significance of Mallory body formation. AB - BACKGROUND/AIMS: Chronic intoxication of mice with 3,5-diethoxycarbonyl-1,4 dihydrocollidine (DDC) or griseofulvin (GF) results in appearance of Mallory bodies (MBs) and alterations of the keratin cytoskeleton, which are reversible upon drug withdrawal but recur after readministration within 2-3 days. METHODS: DDC- or GF-treated and recovered mice were reintoxicated with the original drugs but also colchicine and lumicolchicine. Cytoskeletal alterations of hepatocytes and MB formation were monitored by immunofluorescence microscopy using keratin, MB-specific antibodies, antibodies to phosphoepitopes and to HSP70. Keratin 8/18 mRNA expression and protein levels were determined by competitive reverse transcription-polymerase chain reaction, in situ-hybridization and western blotting. RESULTS: Duration of pretreatment was important for the efficiency of MB triggering. Rapid increase of keratin 8/18 mRNA and proteins was found in all reintoxicated mice concomitant with MB formation, whereby keratin 8 prevailed over keratin 18. Keratins and a protein with heat shock characteristics (M(M) 120 1 antigen) were the earliest detectable MB components, whereas ubiquitination and phosphorylation followed later. CONCLUSIONS: Overproduction of keratins is a major but not the only step responsible for MB formation. Additional components (e.g. M(M) 120-1 antigen) and excess of keratin 8 over keratin 18 are essential. PMID- 11434613 TI - Mechanism of novel vitamin K analog induced growth inhibition in human hepatoma cell line. AB - BACKGROUND/AIMS: To understand the mechanisms of liver regeneration or hepatoma apoptosis, it is important to estimate the turning point of the signal transduction by growth factor receptor. Since 2-(2-hydroxyethylsulfaryl) 3-methyl 1,4-naphthoquinone or CPD 5 has been shown to mediate the phosphorylation of epidermal growth factor (EGF) receptor in Hep3B hepatoma cells, the differences between EGF and CPD 5-mediated signal transduction were studied. METHODS: DNA content was measured by Hoechst fluorescent assay. Phosphorylated proteins were described with Western blots or two-dimensional electrophoresis. RESULTS: CPD 5 induced EGFR phosphorylation was functional to stimulate Ras pathway. However, CPD 5-mediated extracellular signal-regulated kinase (ERK) phosphorylation was not antagonized by inhibition of upstream activation with PD153035. CPD 5 inhibited ERK dephosphorylation in cell lysate, suggesting that ERK phosphorylation by CPD 5 was depending on kinase activity and phosphatase inhibition. Two-dimensional electrophoresis showed extra phospho ERK spot, which was indicated to have close association with CPD 5-induced growth inhibition, since U0126 antagonized growth inhibition and appearance of this spot. CONCLUSIONS: The turning point of EGFR pathway was proved to have close association with the expressed level of phosphorylated ERK. ERK phosphorylation was suggested to play a critical role in growth factor-induced signal transduction. PMID- 11434614 TI - Vascular endothelial growth factor secreted by replicating hepatocytes induces sinusoidal endothelial cell proliferation during regeneration after partial hepatectomy in rats. AB - BACKGROUND: The aim of this study was to investigate regulatory mechanisms of sinusoidal endothelial cell (SEC) proliferation after hepatectomy in rats. METHODS: We investigated expressions of vascular endothelial cell growth factor (VEGF) and its receptors, flt-1 and KDR/flk-1, in regenerating liver after 70% hepatectomy. Proliferation of both hepatocytes and SECs was also monitored by evaluating the proliferating cell nuclear antigen (PCNA) labeling index. Furthermore, VEGF production by cultured hepatocytes isolated at different times after hepatectomy was measured in vitro. RESULTS: The expression of VEGF mRNA was increased markedly between 48 and 72 h after hepatectomy, and thereafter decreasing gradually. The immunohistochemical staining revealed that expression of VEGF started to increase 24 h after hepatectomy, with a peak at 72 h, and the majority of the VEGF-positive cells were hepatocytes located in periportal areas. Meanwhile, expression of flt-1 and KDR/flk-1 was observed along the sinusoids even before hepatectomy, but was increased between 72 and 120 h. Furthermore, VEGF production by cultured hepatocytes isolated 72 h after hepatectomy was significantly increased. The PCNA labeling index of the SECs exhibited a delayed and slower regenerative response in comparison to the hepatocytes, reaching a peak at 72 h. CONCLUSIONS: These data strongly suggest that VEGF secreted by proliferating hepatocytes may represent an important stimulator of SEC proliferation. PMID- 11434615 TI - Sinusoidal endothelial cell proliferation and expression of angiopoietin/Tie family in regenerating rat liver. AB - BACKGROUND/AIMS: Angiogenesis is essential in liver regeneration. However, only little is known about sinusoidal endothelial cell proliferation and the role of different angiogenic growth factors and their receptors during regeneration. METHODS: Seventy percent hepatectomy was carried out on male rats. Serial changes in endothelial cell proliferation were evaluated by immunohistochemistry. The mRNA expressions of angiogenic growth factors (vascular endothelial growth factor (VEGF) and angiopoietins 1 and 2) and their receptors (flt-1, flk-1, Tie-1 and Tie-2) in the whole liver were evaluated by semi-quantitative RT-PCR. RESULTS: Significant elevation of endothelial cell proliferation started at 48 h and peaked at 72 h after hepatectomy. The ratio of sinusoids to liver tissue area initially decreased at 72 h, and thereafter, significantly increased at 96 h. VEGF related factors had early peaks, which coincided with the endothelial proliferation. flt-1, flk-1 and VEGF expressions peaked at 24, 48 and 72 h, respectively. angiopoietin/Tie factors peaked at 96 h, except Ang-2, which gradually increased and peaked at 168 h. CONCLUSIONS: During liver regeneration, hepatocyte proliferation was followed by endothelial cell proliferation. The VEGF family and angiopoietin/Tie system may have distinct roles in angiogenesis, with an enhanced expression of the VEGF family in the early phase of regeneration followed by angiopoietin/Tie expression. PMID- 11434616 TI - Foetal rise in hepatic enzymes follows decline in c-met and hepatocyte growth factor expression. AB - BACKGROUND/AIMS: In the embryo, rapidly proliferating hepatocytes migrate from the liver primordium into the surrounding mesenchyme, whereas foetal hepatocytes are mitotically quiescent and accumulate hepatocyte-specific enzymes. We investigated the timing and topography of this behavioural switch. METHODS: The expression of the c-met receptor and its ligand, hepatocyte growth factor (HGF), was investigated in prenatal rat liver by in situ hybridization, immunohistochemistry and western-blot analysis. RESULTS: c-Met was expressed by hepatocytes and HGF by non-parenchymal liver cells. Their mRNA levels peaked during embryonic day (ED) 11-13. c-Met protein was weakly expressed in the entire liver during ED 11 and 12, but more abundantly at ED 13, when its expression withdrew to the hepatic periphery. Simultaneously, the periportal hepatocellular marker carbamoylphosphate synthetase began to accumulate in the centre of the liver. Although the definitive vascular architecture develops simultaneously, the downstream, pericentral hepatocytes began to express glutamine synthetase only 4 days later, suggesting a requirement for prior periportal hepatocyte maturation. Additionally, c-met protein appeared in the connective tissue surrounding the large veins. The c-met protein/mRNA ratio was substantially higher in non epithelial cells (hepatic connective tissue, heart) than in endoderm-derived epithelia, including hepatocytes, indicating important post-transcriptional regulation. CONCLUSIONS: The decline in c-met expression reflects the end of the embryonic phase and heralds the onset of the fetal, maturational phase of liver development. PMID- 11434617 TI - Recipient HLA-DR3, tumour necrosis factor-alpha promoter allele-2 (tumour necrosis factor-2) and cytomegalovirus infection are interrelated risk factors for chronic rejection of liver grafts. AB - BACKGROUND/AIMS: The tumour necrosis factor (TNF)-2 promoter allele, which elicits elevated expression of TNF-alpha, is in linkage disequilibrium with the extended haplotype HLA-A1-B8-DR3-DQ2. TNF-2 and HLA-DR3 have been implicated in renal and cardiac graft rejection and loss. Cytomegalovirus (CMV) infection has been associated with chronic allograft rejection. We examined the relationship between HLA-DR3, promoter allele TNF-2 and cytomegalovirus in relation to chronic rejection following liver transplantation. METHODS: (i) Retrospective analysis of HLA-DR3 was performed in 307 liver transplant recipients and 283 donors. (ii) Prospective analysis of TNF-alpha promoter allele status, HLA-DR3 status and cytomegalovirus infection was assessed in 123 recipients. RESULTS: (i) Retrospective analysis. Recipient HLA-DR3 (relative risk 1.9; 95% C.I. 1.01-3.58) was a risk factor for chronic rejection. (ii) Prospective analysis. Recipient HLA DR3 was a risk factor for chronic rejection (relative risk 3.41; 95% C.I. 1.66 7.03) which was elevated further by superimposed CMV infection (relative risk 5.01; 95% C.I. 2-12.55). Recipient TNF-2 was associated with chronic rejection (relative risk 2.29; 95% C.I. 0.9-5.83) through linkage to HLA-DR3. CONCLUSIONS: Recipient HLA-DR3, TNF-2 status and CMV infection were inter-related risk factors for chronic rejection of liver grafts. PMID- 11434618 TI - Histological features after liver transplantation in alcoholic cirrhotics. AB - BACKGROUND/AIMS: Though alcoholic cirrhosis is a common indication for liver transplantation, it carries the risk of alcohol recidivism and consequent graft failure. This study aims to evaluate the effect of alcohol recidivism on survival rates and histological parameters in patients transplanted for alcoholic cirrhosis, with and without hepatitis C virus (HCV) infection. METHODS: Fifty-one out of 189 consecutive transplanted patients underwent psychosocial evaluation and liver biopsy at 6 and 12 months, then yearly after transplantation. RESULTS: The cumulative 84 month survival rate was identical in patients transplanted for alcoholic (51%) and non-alcoholic cirrhosis (52%). No difference emerged between anti-HCV negative vs. positive alcoholic cirrhosis patients. Psycho-social evaluation revealed alcohol recidivism in 11/34 long-term survivors, but this did not affect overall survival rate in patients with or without HCV. In anti-HCV negative cases, fatty changes and pericellular fibrosis were significantly more common in heavy drinkers than in occasional drinkers and abstainers. When HCV status was considered regardless of alcohol intake, fibrosis was significantly more frequent in patients with HCV. CONCLUSION: Alcohol recidivism after transplantation in alcoholic cirrhosis patients does not affect survival, irrespective of HCV status. Fatty changes and pericellular fibrosis are the most relevant histological signs of heavy alcohol intake. PMID- 11434619 TI - High levels of soluble tumor necrosis factor superfamily receptors in patients with hepatitis C virus infection and lymphoproliferative disorders. AB - BACKGROUND: Chronic hepatitis C virus (HCV) infection is associated with a variety of extrahepatic disorders that may relate to direct or indirect effects of virus infection. Increased levels of soluble forms of tumor necrosis factor (TNF) receptors I and II, found in lymphoproliferative and infectious diseases, can interfere with TNF induced apoptotic cell death. The aim of the present study was to evaluate soluble TNF family receptors levels in lymphoproliferative disorders associated with HCV infection. METHODS: One hundred and forty-nine subjects were studied, including 120 anti-HCV positive patients (60 without lymphoproliferative manifestations, 47 with type II cryoglobulinemia and 13 with low-grade B-cell non-Hodgkin's lymphoma (B-NHL)) and 29 anti-HCV negative subjects (19 with low-grade B-NHLs and ten normal controls). RESULTS: Soluble forms of TNF receptor I, TNF receptor II and Fas were significantly higher in HCV positive patients compared with normal controls. The highest levels were found in patients affected by type II cryoglobulinemia or HCV positive lymphoplasmacytoid lymphomas (LP-NHLs), while HCV positive patients without type II cryoglobulinemia or with other B-NHLs had lower values (P < 0.01). CONCLUSIONS: Among HCV infected individuals, very high levels of soluble TNF receptors are significantly associated with type II cryoglobulinemia and LP-NHLs, suggesting that they may be involved in these proliferative disorders. PMID- 11434620 TI - Rates and risk factors of liver fibrosis progression in patients with chronic hepatitis c. AB - BACKGROUND AIMS: In hepatitis C there is controversy over the linearity of the rate of progression and the significance of gender, mode of infection and viral factors. METHODS: 2313 untreated patients with a reliable estimated duration of infection and liver fibrosis were included. Fibrosis progression was calculated using the Kaplan-Meier method and the rate of fibrosis progression using the hazard function. Seven risk factors were assessed: age at biopsy, gender, alcohol consumption, mode of infection, activity grade, hepatitis C virus genotype and RNA level. RESULTS: The percentage of patients without cirrhosis was 91% after 20 years of infection (95% CI:90-92%) and 56% after 40 years (95% CI:48-64%). Three independent factors were associated (P < 0.001) with a faster progression rate: age at infection, alcohol consumption of 50 g or more per day, and male gender. The mode of infection, histologic activity, genotype and viral load were not independently associated with fibrosis. Fibrosis progression was mainly dependent on age and the duration of infection and can be divided into four successive periods with very slow, slow, intermediate and rapid progression rates. CONCLUSION: In patients infected with hepatitis C, the majority of fibrosis progression occurred in those aged fifty years or older. PMID- 11434621 TI - Liver fibrosis assessment with semiquantitative indexes and image analysis quantification in sustained-responder and non-responder interferon-treated patients with chronic hepatitis C. AB - BACKGROUND/AIMS: The effect of interferon on the reduction of liver fibrosis is controversial. We aimed to compare semiquantitative methods with a quantitative digital image analysis system to assess liver fibrosis in biopsies from patients with chronic hepatitis and different responses to interferon. METHODS: We studied 98 liver biopsies with chronic hepatitis C before and after recombinant interferon alfa-2 treatment, using conventional histological assessment, grading of histological activity, scoring/staging of fibrosis (Knodell and Scheuer), and quantification of fibrosis with image analysis (FibroQuant). RESULTS: Sustained responders to interferon showed a significant reduction in histological lesions and in their Knodell and Scheuer activity indexes. The semiquantitative systems showed no reduction in fibrosis. The FibroQuant application showed a significant reduction in porto-periportal and septal areas among sustained-responders (P < 0.001) and non-responders (P < 0.05), and in porto-periportal and septal fibrosis areas only in sustained-responders (P < 0.001), whereas the percentage of fibrosis increased in non-responders (P < 0.001). CONCLUSIONS: The Scheuer system is useful for the daily evaluation of fibrosis, but the FibroQuant application provides more objective data on the anti-fibrogenic effects of interferon, which include a reduction in the porto-periportal area in sustained-responders and non responders, accompanied by a reduction in the area of fibrosis only when the viral replication has ceased. PMID- 11434622 TI - Long-term beneficial effects in sustained responders to interferon-alfa therapy for chronic hepatitis C. AB - BACKGROUND/AIMS: Assessment of chronic hepatitis C outcome in sustained responders to interferon requires prolonged observation and close monitoring. We prospectively studied the impact of sustained response on histology and clinically relevant outcomes. METHODS: The 47 sustained responders (ten with cirrhosis) from two interferon trials involving 235 chronic hepatitis C patients (81 with cirrhosis) were included. Hepatitis C virus (HCV) RNA was assessed every 6 months, liver histological changes from baseline, 6-12 and 48-72 months after treatment discontinuation. RESULTS: The mean follow-up was 102 +/- 19 months. HCV RNA became undetectable in 36/47 responders. Four responders, who had remained viremic, later relapsed. The histology progressively improved in non-viremic and viremic patients, with a more marked improvement in the former (P = 0.0089), normalizing in 53 vs. 0% (P = 0.0220). No patient progressed to cirrhosis. One non-viremic cirrhotic patient developed a hepatocellular carcinoma. Non responders from the two original trials had worse histological outcomes and those with cirrhosis had a higher rate of clinically relevant events compared with cirrhotics showing a sustained biochemical response (4.5 vs. 1.2 cases/100 person years; CI for the difference, 0.3-6.3). CONCLUSIONS: Most sustained, virological responders without cirrhosis normalize liver histology in the long-term and are cured of the disease. Sustained responders remaining viremic still show histological improvement, albeit to a lesser extent. PMID- 11434623 TI - Caveats concerning hepatopulmonary syndrome. PMID- 11434624 TI - Vascular endothelial growth factor and liver regeneration. PMID- 11434625 TI - Serial liver biopsies: a gateway into understanding the long-term health of the liver allograft. PMID- 11434626 TI - Progression of fibrosis in chronic liver diseases: time to tally the score. PMID- 11434627 TI - Neuropsychological characterization of hepatic encephalopathy. PMID- 11434628 TI - Successful combination therapy of polyarteritis nodosa associated with a pre-core promoter mutant hepatitis B virus infection. AB - BACKGROUND/AIMS: To describe the clinical and virological evolution of a polyarteritis nodosa (PAN) case associated with a hepatitis B virus (HBV) pre core promoter mutant infection that was successfully treated with plasma exchanges, corticosteroids, and interferon alpha (IFN-alpha). METHODS: Viral markers were used, including HBV DNA quantified by the branched DNA assay and detected by PCR, the HBV genome sequence, pre-S1Ag and anti-HBC IgM which were studied throughout the treatment period and the entire follow-up in the serum, while the presence of virus in extrahepatic sites was detected by immuno staining. RESULTS: The patient was infected with a typical pre-core promoter mutant harboring four point mutations. Pre-S1Ag was cleared rapidly from serum, most likely via the formation of immune complexes since HBV DNA declined more progressively. Viral infection was then cleared after a second episode of hepatocyte lysis. This was accompanied by a recovery from all clinical manifestations. CONCLUSIONS: The favorable treatment outcome observed in this first case of pre-core promoter HBV mutant associated PAN underlines that combination therapy based on IFN-alpha can clear pre-core promoter HBV infection and cure PAN. It also provides new insight in the pathogenesis of HBV associated PAN. PMID- 11434629 TI - Images in hepatology. Aneurysm of the trunk of the portal vein. PMID- 11434630 TI - Liver nodules resembling focal nodular hyperplasia. PMID- 11434631 TI - Normal red cell cardiac output in the hyperkinetic syndrome of alcoholic cirrhosis. PMID- 11434632 TI - Efavirenz-induced acute eosinophilic hepatitis. PMID- 11434633 TI - Antibodies to Clp protease in primary biliary cirrhosis: possible role of a mimicking T-cell epitope. PMID- 11434634 TI - Ursodeoxycholic acid for primary biliary cirrhosis: lesson for the future? PMID- 11434635 TI - Values and assumptions in the development of DSM-III and DSM-III-R: an insider's perspective and a belated response to Sadler, Hulgus, and Agich's "On values in recent American psychiatric classification". AB - It is widely acknowledged that the approach taken in the development of a classification of mental disorders is guided by various values and assumptions. The author, who played a central role in the development of DSM-III (American Psychiatric Association [1980] Diagnostic and statistical manual of mental disorders, 3rd ed. Washington, DC:Author) and DSM-III-R (American Psychiatric Association [1987] Diagnostic and statistical manual of mental disorders, 3rd ed, rev. Washington, DC:Author) will explicate the basic values and assumptions that guided the development of these two diagnostic manuals. In so doing, the author will respond to the critique of DSM-III and DSM-III-R made by Sadler et al. in their 1994 paper (Sadler JZ, Hulgus YF, Agich GJ [1994] On values in recent American psychiatric classification. JMed Phil 19:261-277). The author will attempt to demonstrate that the stated goals of DSM-III and DSM-III-R are not inherently in conflict and are easily explicated by appealing to widely held values and assumptions, most of which appeared in the literature during the development of the manuals. Furthermore, we will demonstrate that it is not true that DSM-III places greater emphasis on reliability over validity and is covertly committed to a biological approach to explaining psychiatric disturbance. PMID- 11434636 TI - The comprehensive assessment of defense style: measuring defense mechanisms in children and adolescents. AB - This study introduces the Comprehensive Assessment of Defense Style (CADS), a new method to assess descriptively the defensive behavior of children and adolescents. Parents of 124 children and adolescents referred to a mental health clinic, of 104 nontreated children, and of 15 children whose fathers were treated for posttraumatic stress disorder completed the CADS. Factor analysis of 28 defenses yielded one mature factor, one immature factor of defenses expressed in relations with the environment (other-oriented), and one of defenses expressed in relations with the self (self-oriented). The CADS significantly discriminated between patients and nonpatients. Psychiatric patients used more immature and fewer mature defenses than control subjects, and adolescents used more mature and fewer other-oriented defenses than children. Girls used more mature and fewer other-oriented defenses than boys. The reliability and validity data of the CADS are encouraging. The three defense factors may be implemented for diagnostic and clinical purposes as well as for screening for psychopathology risk in untreated populations. PMID- 11434637 TI - Validation of the Comprehensive Assessment of Defense Style (CADS): mothers' and children's responses to the stresses of missile attacks. AB - This study furthers the validation of the Comprehensive Assessment of Defense Style (CADS) as a measure of children's defensive behavior. Participants were 81 mothers who assessed the defense style (CADS) of their 8- to 10-year-old children, as well as their own defense style and level of object relations. Five years earlier, the mothers had rated their children's symptom level and personality after the missile attacks during the Gulf War. The original factor structure of the CADS was replicated for the most part. Self-oriented and other oriented defenses were related to the children's early personality and symptomatic reaction, as well as to their mother's defense style and level of object relations. The CADS factors correlated with the defenses of the Defense Mechanisms Manual. The results provide further validation of the CADS and suggest possible areas of implementation, such as longitudinal examination of defenses, psychopathology screening, and therapeutic improvement. PMID- 11434638 TI - The effect of institutional fiscal stress on the use of atypical antipsychotic medications in the treatment of schizophrenia. AB - This paper examines the effects of medical center budget stress on the use of expensive atypical antipsychotic medications for the treatment of schizophrenia in the Department of Veterans Affairs (VA). VA prescription drug records were collected for patients diagnosed with schizophrenia. Generalized estimation equations were used to identify patient and facility characteristics (especially fiscal stress) that are associated with the use of atypical antipsychotics. Of the 34,925 patients in the final sample, over half received an atypical antipsychotic, usually either olanzapine or risperidone. Unexpectedly, increased fiscal stress was associated with increased likelihood of receiving atypical antipsychotics. Among patients who receive atypicals, however, fiscal stress was associated with reduced likelihood of receiving the more expensive atypicals (clozapine and olanzapine) but positively associated with receiving the least expensive atypical (risperidone). Institutional fiscal pressure does not seem to reduce the broad availability of these medications overall but does affect which drug is prescribed. PMID- 11434639 TI - Drug treatment outcomes for adolescents with comorbid mental and substance use disorders. AB - This study compared the pretreatment characteristics and posttreatment outcomes of substance-abusing adolescents with and without comorbid mental disorders in the Drug Abuse Treatment Outcome Studies for Adolescents. Subjects (N = 992) were sampled from 23 adolescent drug treatment programs across three modalities (residential, short-term inpatient, outpatient drug-free). Nearly two thirds (64%) of the sample had at least one comorbid mental disorder, most often conduct disorder. Comorbid youth were more likely to be drug or alcohol dependent and had more problems with family, school, and criminal involvement. Although comorbid youth reduced their drug use and other problem behaviors after treatment, they were more likely to use marijuana and hallucinogens, and to engage in illegal acts in the 12 months after treatment, as compared with the noncomorbid adolescents. Integrated treatment protocols need to be implemented within drug treatment programs in order to improve the outcomes of adolescents with comorbid substance use and mental disorders. PMID- 11434640 TI - Suprathreshold repetitive transcranial magnetic stimulation elevates thyroid stimulating hormone in healthy male subjects. AB - Repetitive transcranial magnetic stimulation (rTMS) has been introduced as a new antidepressive treatment strategy. The mode of action by which the antidepressive effect is brought about is not yet clear. Other antidepressive treatment strategies such as sleep deprivation are associated with an increase of plasma thyroid-stimulating hormone (TSH) levels that correlate with clinical improvement. In the present study, the effect of left prefrontal suprathreshold (120% of motor threshold) rTMS on TSH plasma levels of 19 healthy male subjects was investigated in comparison with subthreshold (80% of motor threshold) and sham stimulation. Suprathreshold rTMS was followed by a significant relative increase of TSH levels 10 and 60 minutes after stimulation in comparison with subthreshold and sham stimulation. The more pronounced effect of suprathreshold rTMS on TSH plasma levels might be important for the determination of optimal stimulation parameters in the treatment of depressed patients. PMID- 11434641 TI - Insight is greater in mixed than in pure manic episodes of bipolar I disorder. PMID- 11434642 TI - Evaluation of a portable low cost videophone system in the assessment of depressive symptoms and cognitive function in elderly medically ill veterans. PMID- 11434643 TI - Self-awarensss in schizophrenia: its relationship to depressive symptomatology and broad psychiatric impairments. PMID- 11434644 TI - Delayed posttraumatic stress disorder (PTSD) and predictors of first onset of PTSD in patients with anxiety disorders. PMID- 11434645 TI - A new century: a new beginning. PMID- 11434646 TI - Most infectious complications in parenterally fed trauma patients are not due to elevated blood glucose levels. AB - OBJECTIVE: To determine the relationship between hyperglycemia and infectious complications in nutritional studies of trauma patients. METHODS: Retrospective review of serum glucose values in two published randomized, prospective studies of patients receiving either enteral or parenteral feeding (trial 1) or isonitrogenous, isocaloric enteral diets (trial 2). Trial 2 also included patients prospectively followed who received little or no enteral feeding. RESULTS: Patients randomized to enteral or parenteral feeding in trial 1 exhibited no significant differences in the highest recorded serum glucose (SG) until the fourth or fifth day after protocol entry. SG tended to be higher in infected than non-infected patients in the first 4 hospital days, but SG was far below values considered to increase the risk for infection (SG > 220 mg/dL). In trial 2, glucose levels tended to be slightly higher in infected than in noninfected patients within the first 5 days reaching statistical significance by day 5. Unfed control patients had similar SG values but significantly more major infectious complications. CONCLUSIONS: Patients developing infections had slightly higher SG levels than noninfected patients early in admission, but these SG values were far below levels considered a risk for infective complications. Significant hyperglycemia does not explain differences in infectious complications in critically ill trauma patients randomized to various routes and types of nutrition. PMID- 11434647 TI - Hyperglycemia and infection: which is the chicken and which is the egg? PMID- 11434648 TI - Metabolic effects of arginine addition to the enteral feeding of critically ill patients. AB - BACKGROUND: Some studies have suggested that the addition of arginine to enteral feeding solutions may improve outcome in critically ill patients, but the mechanism is incompletely explained. In particular, the availability and utilization of arginine administered enterally is not well defined. METHODS: This prospective, randomized, double-blind, placebo-controlled study performed in a Department of Medicosurgical Intensive Care included 51 patients likely requiring long-term enteral feeding. Thirty-seven patients (57 +/- 7 years, SAPS II 33 +/- 6) completed the 7-day study, of whom 20 received the formula enriched with free arginine (6.3 g/L) and 17 received an isocaloric and isonitrogenous control solution. Arginine absorption was assessed from plasma arginine concentrations in serial samples. Three pathways of arginine utilization were explored: (1) the production of nitric oxide, assessed by the plasma concentration of nitrite/nitrate (NOx) and citrulline, and 24-hour urinary excretion of NOx; (2) the protein turnover, estimated by the phenylalanine concentrations; and (3) the activity of arginase, reflected by the ornithine concentration. RESULTS: The plasma concentrations of arginine and ornithine increased in the group fed with the enriched formula (from 55 +/- 9 micromol/L to 102 +/- 9 micromol/L and from 57 +/- 7 to 135 +/- 11 micromol/L, respectively, p < .05), but not with the control formula. There was no difference between groups in either NO production or phenylalanine concentration. CONCLUSIONS: Supplemental arginine in enteral feeding is readily absorbed, and mainly metabolized into ornithine, presumably by the arginase enzyme. PMID- 11434649 TI - Sites of conditional essential fatty acid deficiency in end stage liver disease. AB - BACKGROUND: End stage liver disease (ESLD) is a devastating illness. Its protean manifestations involve many different aspects of disturbed hepatic function. One consequence of ESLD is a decrease in plasma levels of very long chain polyunsaturated fatty acids (VL-PUFAs), particularly arachidonic acid (AA) and docosahexaenoic acid (DHA), the former important for eicosanoid metabolism and the latter for retinal and brain membrane structure. The purpose of this study was to define the VL-PUFA changes in liver disease by comparing plasma and tissue levels of VL-PUFAs in controls to patients with ESLD. METHODS: Fatty acid profiles from plasma, red blood cell (RBC) membranes, muscle, liver, and fat tissue from ESLD patients undergoing liver transplants were measured and compared with control patients undergoing elective liver resection. RESULTS: Fatty acid profiles from plasma and RBC membranes showed significant decreases in AA and DHA levels in patients with ESLD compared with controls. However, there were no significant differences in tissue fatty acid composition between ESLD patients and controls. CONCLUSIONS: ESLD affects the liver's ability to maintain circulating levels of AA and DHA, and thereby presumably RBC membrane levels. However, solid tissues appear not to be affected by ESLD. Although the mechanism for these changes remains to be defined, it is consistent with hepatic impairment of elongation and desaturation to produce VL-PUFA for transport. The present results also suggest that dietary interventions to include preformed VL-PUFA rather than their precursors, linoleic and alpha linolenic acid, would be needed to normalize plasma VL-PUFA levels in patients with ESLD. PMID- 11434650 TI - Oral arginine improves blood pressure in renal transplant and hemodialysis patients. AB - BACKGROUND: Hypertension in kidney transplant (KT) patients may result from attenuated whole-body nitric oxide (NO) content and abnormal NO-mediated vasodilation. Increasing NO bioavailability with L-arginine (ARG) could theoretically restore the NO-mediated vasodilatory response and lower blood pressure. METHODS: In a prospective pilot study, 6 normotensive volunteers and 10 KT patients received oral supplements of ARG (9.0 g/d) for 9 days, then 18.0 g/d for 9 more days. Six hemodialysis (HD) and 4 peritoneal dialysis patients received the same dose for 14 days. Five KT patients received 30 mL/d of canola oil (CanO) in addition to ARG. Systolic (SBP) and diastolic (DBP) blood pressure, creatinine clearance (CCr), and serum creatinine (Cr) were measured at baseline, day 9, and day 18. In a subsequent study, 20 hypertensive KT patients with stable but abnormal renal function were randomized in a crossover study to start ARG only or ARG+CanO supplements for two 2-month periods with an intervening month of no supplementation. SBP, DBP, CCr, and Cr were measured monthly for 7 months. RESULTS: In the pilot study, ARG reduced the SBP in HD patients from 171.5 +/- 7.5 mmHg (baseline) to 142.8 +/- 8.3 mmHg (p = .028). In the crossover study, SBP was reduced from baseline (155.9 +/- 5.0 mmHg), after the first 2 months (143.2 +/- 3.2 mmHg; p = .03) and subsequent 2 months (143.3 +/- 2.5 mmHg; p = .014) of supplementation. DBP was also reduced after supplementation in both studies. CanO had no effect on blood pressure. Renal function did not change. CONCLUSIONS: Oral preparations of ARG (+/-CanO) were well tolerated for up to 60 consecutive days and had favorable effects on SBP and DBP in hypertensive KT and HD patients. PMID- 11434651 TI - Outcome of patients treated with home enteral nutrition. AB - BACKGROUND: The aims of this study were to prospectively analyze the 1-month mortality and long-term outcome of home enteral nutrition (HEN) patients in order to determine the benefits of this treatment. METHODS: Between 1990 and 1996, 417 patients, aged 64 +/- 25 years, were discharged on HEN and followed up until December 31, 1998, when outcome was assessed, which allowed us to determine survival probabilities and conditions associated with survival. RESULTS: The mean duration of HEN was 242 +/- 494 days, with a 24- to 103-month follow-up. Probabilities of being alive at 1 month, 1 year, and 5 years were 80%, 41.7%, and 25%, respectively. Factors associated with death were dementia, neurologic disease, head and neck cancer, AIDS, and age over 70 years. A total of 5.5% of patients remained dependent on HEN, 32.6% resumed full oral nutrition, 20.2% of patients died during the first month on HEN, and 35% died after more than 1 month on HEN (219 +/- 257 days). A total of 6.7% of patients stopped HEN for other reasons. CONCLUSIONS: HEN provides well-tolerated long-term nutritional support in many patients. However, because of their likelihood of being old and the nature of the underlying disease, these patients as a group tend to have a modest prognosis. This calls for the determination of more accurate selection criteria, and the measurement of the impact of HEN on quality of life. PMID- 11434652 TI - A controlled comparison of traditional feeding tube verification methods to a bedside, electromagnetic technique. AB - BACKGROUND: Benefits of enteral feeding are diminished by aspiration pneumonia and mechanical complications of misplaced feeding tubes. To avoid complications, clinicians determine the location of the tip before feeding. This study compares diagnostic test characteristics of 4 techniques for tip localization. METHODS: A prospective, blinded trial was conducted on the wards and critical care units of four acute-care hospitals. Patients requiring at least 3 days of enteral feeding were studied. Four observers at each institution used a randomly assigned technique to determine a tube's tip location. Methods included auscultation, aspiration with inspection or pH determination of aspirated material, and a recently developed electromagnetic technique. Results were compared with radiographic determination. Success rates were compared using sensitivity, specificity, and likelihood ratios. RESULTS: The 4 methods agreed with the radiograph in (mean, 95% confidence interval): 84 (80 to 88)%, 50 (45 to 55)%, 56 (51 to 61)%, and 76 (72 to 81)% of observations, respectively. Only the electromagnetic method and aspiration identified all tubes located above the diaphragm (negative likelihood ratio 0 and sensitivity 100%). Aspiration was unsuccessful in making a determination in 53% of the observations, whereas the electromagnetic method was successful 90% of the time. PMID- 11434653 TI - Pyruvate in organ transplantation. AB - Pyruvate (PY) is a 3-carbon compound present in human tissues and physiologically used by cells as an energetic substrate in anaerobic conditions. In the last few years, we have successfully used PY to protect small bowel (SB), liver, and kidneys from ischemia/reperfusion injury in several experimental models. Although the mechanism of protection is not fully clarified, we have shown increased tissue levels of adenosine triphosphate (ATP) during ischemia. This suggests that providing supra-physiologic concentrations of PY during anaerobic glycolysis might enable the cells to remain viable during prolonged hypoxia. Furthermore, mechanisms such as direct inhibition of oxygen free radical formation, abrogation of neutrophilic infiltration and reduced up-regulation of adhesion molecules have also been documented in these studies. In light of these findings, we evaluated the efficacy of PY in organ preservation and transplantation. We demonstrated a protective effect on intestinal preservation injury and during acute rejection. Oral PY treatment induced immunologic changes in rejecting allograft, inhibiting perforin and granzyme-b expression and leukocytic infiltration. Protection was also documented on livers after prolonged hypothermic preservation using a PY based preservation solution. Additionally, isolated pancreatic islets were cultured in PY enriched media and maintained viable for up to 120 days followed by in vitro testing and transplantation revealing a well preserved function. All these findings suggest that PY is a potentially beneficial nutrient in patients undergoing organ transplantation and that future clinical application of PY in this field should be encouraged. PMID- 11434654 TI - Nutrient support in hematopoietic cell transplantation. AB - High-dose cytoreduction and hematopoietic stem cell infusion form the basis for treatment of hematologic cancers, defects or failure of hematopoiesis, and some solid tumors. As an antitumor therapy, allogeneic hematopoietic cell transplantation (HCT) is superior to autologous HCT by induction of a graft-vs tumor effect. However, recipients of allografts suffer higher transplant-related mortality owing to graft-vs-host disease (GVHD). Nutrition support research must recognize that HCT is a heterogeneous modality whose short and long-term outcomes are affected by transplant type, preparative regimens, diagnosis, disease stage, age, and nutritional status. The field of HCT will diversify further as lower dose cytoreduction and mixed chimerism grafts allow expansion of the technique to older patients and to other diseases. PMID- 11434655 TI - Kids Welcome Here. PMID- 11434656 TI - Whiplash. PMID- 11434657 TI - Certificate vs. Certification. PMID- 11434658 TI - The visual profile of Romanian children and adults assessed through vision screenings. AB - BACKGROUND: The purpose of this study was to databank epidemiological information from Romanian children and adults. METHODS: A vision screening was performed on 690 children (2 to 18 years of age) and 190 adults (19 to 77 years of age). Prevalence of refractive errors, binocularity, and ocular health problems were recorded and evaluated. RESULTS: Data from the childrens' population showed that 45% were emmetropic, 27% were myopic, and 28% were hyperopic. For the adults, 42% were emmetropic, 16% were myopic, and 42% were hyperopic. When compared with other nations, the average refractive error of the Romanian children was more myopic, and for the adults it was slightly more hyperopic. With-the-rule astigmatism had the highest occurrence when compared to other axis orientations, yet the overall occurrence of astigmatism was less than that found in other nations for both children and adults. The prevalence of strabismus and ocular disease was less in the Romanian children in relation to other nations. CONCLUSION: Due to many years of isolation from the Free World, little information has been available on nations such as Romania relative to the health status of its people. This study begins an epidemiological base forthe nation. It is especially important to public health officials in planning health policy measures, and provides a basis for additional U.S.-based outreach missions to such nations in Europe. It also illustrates the efficacy of using a basic screening-type examination to determine vision care needs and the use of recycled eyewear to satisfy these needs. PMID- 11434659 TI - Operations management. PMID- 11434660 TI - RX for pharmaceutical practice. PMID- 11434661 TI - Thyroid diseases in patients treated during pre-puberty for medulloblastoma with different radiotherapic protocols. AB - We evaluated thyroid disease in 32 patients treated, during pre-puberty, for medulloblastoma, followed for at least 4 years and without relapse during observation. After surgery the patients underwent chemotherapy (CT) and radiotherapy (RT). The protocols were as follows: 20 patients (group A) SNC 76 and SNC 85 protocols which included conventional fractionated RT (36-40 Gy to the craniospinal axis and a 14-18 Gy boost to the posterior fossa, administered as 1.5-1.8 Gy per fraction per day) and a junction between the cranial and the spinal fields at C2-C3 level; 12 patients (group B) SNC 91 protocol which included hyperfractionated RT (36 Gy to the craniospinal axis and a 30 Gy boost to the posterior fossa; this was administred as 1 Gy per fraction twice per day) and a junction at levels C5-C6 or C6-C7 level. The mean age at diagnosis was 7.4+/-3.2 years for group A and 8.4+/-2.6 years for group B. Thyroid function was evaluated yearly and ultrasonographic characteristics every 2 years. The patients were followed for a mean of 10.8+/-3.8 for group A and 6+/-1.4 years for group B. Primary hypothyroidism was diagnosed in 16 group A patients and 4 group B patients, and central hypothyroidism was diagnosed in 2 group A patients (difference in risk of developing hypothyroidism evaluated with a Wilcoxon-test: p=0.048). Ultrasonography showed reduced thyroid volume in 7 group A cases, and structural changes in 21 patients (17 group A, 4 group B); 9 L-thyroxine-treated patients were confirmed hypothyroid after having stopped therapy. A thyroid nodule was detected in two cases (one from each group). In conclusion, our data indicate that thyroid injury may be diminished by the use of hyperfractionation and low-junction radiotherapy in the treatment of medulloblastoma. PMID- 11434662 TI - Changes in motor control and muscle performance after a short-term body mass reduction program in obese subjects. AB - Two hundred and thirty obese subjects (age: 18-77 yr, BMI: 31.1-65.8 kg/m2) were studied before and after a 3-week body mass reduction (BMR) program, coupling restricted energy diet (1200-1500 kcal/day) with low intensity exercise prescription. It involved 5 days per week (consisting of one-hour dynamic aerobic standing and floor exercise plus 30 min of cycloergometer exercise at 60 W or, alternatively, 4 km outdoor leisure walking on flat terrain) and psychological counseling. One-leg standing balance test (OLSB) and stair climbing test (SCT) were employed to assess motor control and maximal lower limb muscle power, respectively. The BMR program induced a significant weight loss (4.1%; p<0.001), a higher reduction of body mass index (BMI) being observed in males than in females (p<0.001). OLSB performance time increased by 20.5% (p<0.001) after treatment, the improvement being evident in both genders. A 20.8% reduction in SCT time (p<0.05) was also observed and corresponded to a 13.2% increase (p<0.001) in average absolute muscle power and 15.0% increase (p<0.001) in specific muscle power (i.e. the power output per kg of body mass), with no differences between genders. In conclusion, in spite of the moderate reduction of body mass after restricted energy diet and low intensity physical conditioning, significant improvements in motor control and performance, likely to ameliorate the execution of simple daily activities, were observed in obese subjects. PMID- 11434663 TI - Inhibition of 5alpha-reductase enzyme or GABA(A) receptors in the VMH and the VTA attenuates progesterone-induced sexual behavior in rats and hamsters. AB - Progesterone (P) to the ventromedial hypothalamus (VMH) and the ventral tegmental area (VTA) of ovariectomized (OVX), estradiol benzoate (EB)-primed rats and hamsters produces female sexual behavior similar to that seen in proestrous, receptive rodents. Because P's 5alpha-reduced metabolites can have facilitative effects on female sexual receptivity through actions at GABA(A)/benzodiazepine receptor complexes (GBRs), the role of 5alpha-reductase and GBRs in the VMH and the VTA was investigated. In Experiment 1, 5alpha-reductase immunoreactivity (5alpha-red-IR) and GBR immunoreactivity (GBR-IR) in the VMH and the VTA of OVX, EB (10 microg) and P (500 microg)-primed rats and hamsters was examined. More 5alpha-red-IR and GBR-IR was seen in the VMH and the VTA of receptive (EB and P primed) compared to non-receptive (sesame oil vehicle) rodents. In Experiment 2, OVX, EB and P-primed rats and hamsters received implants of finasteride, a 5alpha reductase inhibitor, or no implants to the VMH and the VTA and were tested for sexual receptivity with a male. Ovariectomized EB and P-primed rats and hamsters receiving finasteride to the VMH and the VTA had decreased lordosis compared to rodents receiving control implants to the VMH and the VTA. In Experiment 3, OVX, EB and P-primed rats and hamsters received infusions of picrotoxin, a GBR antagonist, or vehicle infusion to the VMH and the VTA and were tested for sexual receptivity with a male. Ovariectomized EB and P-primed rats and hamsters receiving picrotoxin to the VMH and the VTA had decreased lordosis compared to rodents receiving vehicle infusions to the VMH and the VTA. These data suggest that 5alpha-reductase and GBRs are present in the VMH and VTA, and that inhibiting 5alpha-reductase activity or blocking GBRs in the VMH and the VTA attenuates EB+P-primed sexual receptivity of OVX rats and hamsters. PMID- 11434664 TI - Endogenous sex steroid, GH and IGF-I levels in normal elderly men: relationships with bone mineral density and markers of bone turnover. AB - There are studies concerning the association among endogenous sex steroids, growth hormone (GH), insulin-like growth factor-I (IGF-I) and bone mineral density (BMD) in both men and women. However, little is known concerning the association of these parameters with markers of bone turnover in healthy elderly men. We studied the association of BMD (dual energy X-ray absorptiometry of spine, hip and forearm) and markers of bone turnover (bone-specific alkaline phosphatase, serum C-terminal propeptide of type I collagen, and serum osteocalcin reflecting formation, urine deoxypyridinoline and calcium excretion in relation to creatinine excretion reflecting resorption) with endogenous sex steroids, GH and IGF-I in 14 elderly normal men (age range 60-79 years). There was a negative correlation between age and dehydroepiandrosterone sulphate (DHEAS) (r=-0.60, p=0.022) and a positive correlation between GH and IGF-I (r=0.53, p=0.048). Serum estradiol concentrations correlated with BMD at distal 1/3 radius (r=0.41, p=0.1) and mid-radius (r=0.47, p=0.08), and negatively correlated with age (r=-0.45, p=0.09). There was no correlation of estradiol with bone turnover markers, testosterone, free testosterone, DHEAS, GH and IGF-I. Serum GH and IGF-I levels showed no correlation with BMD (all sites) and bone turnover markers. Serum total testosterone concentrations positively correlated with BMD at distal 1/3 radius (r=0.47, p=0.09), femoral neck (r=0.56, p=0.037) and Ward's triangle (r=0.49, p=0.07). These data suggest that serum estradiol and testosterone levels are associated with BMD in elderly men, possibly indicating their contribution to skeletal maintenance in old age. However, correlations of IGF-I, GH and DHEAS with BMD and bone turnover markers are lacking in the group studied. PMID- 11434665 TI - Somatostatin receptor scintigraphy in the follow-up of patients with differentiated thyroid cancer. AB - The aim of this study was to compare the results of somatostatin receptor scintigraphy (SRS) and of radioiodine scintigraphy in patients with metastatic differentiated thyroid carcinoma during L-thyroxine suppression therapy and after withdrawal. Twenty-five patients were studied: 16 patients had papillary cancer and 12 of them had metastatic disease; 9 patients had follicular cancer and 7 of these had known metastases. In 7 patients SRS was performed during thyroxine withdrawal, in 12 during thyroxine therapy within 9 months from radioiodine scintigraphy, in 6 others both during suppression therapy and after withdrawal. SRS was positive in 18 of 25 (72%) patients. It demonstrated lesions in 11 of 13 (85%) patients after thyroxine withdrawal and in 12 of 18 (67%) patients during thyroxine suppression. In 6 patients in whom a direct comparison was made before and after withdrawal, essentially the same information was obtained. Six of 8 (75%) patients with lesions that did not concentrate radioiodine showed uptake of labeled octreotide in these lesions. In 5 of 17 (29%) patients whose tumors concentrated radioiodine, no uptake was found during SRS. CONCLUSIONS: 1) in patients with metastatic differentiated thyroid carcinoma, tumor sites can be visualized using SRS; 2) there is no need to withdraw patients from suppression therapy in order to perform SRS; 3) in some patients whose lesions do concentrate labeled octreotide but not radioiodine, the use of somatostatin analogues labeled with (111)In or [90Y] can provide new therapeutic options. PMID- 11434666 TI - Response of biochemical markers of bone turnover to estrogen treatment in post menopausal women: evidence against an early anabolic effect on bone formation. AB - Bone loss following menopause can be prevented or reduced by estrogen replacement therapy (ERT). The primary action of estrogen on bone is generally considered to be antiresorptive, but some evidence would also suggest a stimulatory effect on bone formation. The aim of this study was to assess the effect of ERT on biochemical markers of bone resorption (urinary pyridinoline and deoxypyridinoline), and of bone formation (bone-specific alkaline phosphatase--B ALP, and the C-terminal propeptide of type I collagen--CICP) in a group of 25 postmenopausal women with no evidence of osteoporosis. Since the suggested anabolic effect of estrogen seems to take place in the early period of ERT, we measured the response of markers immediately before and after the start of treatment (30, 60, 120 and 180 days). The markers of bone resorption started to decrease at 30 days and remained low thereafter. We also observed a similar decrease in serum levels of B-ALP and CICP, reflecting a reduction of bone formation rate. Our data would indicate that ERT at the given dose does not have early anabolic effects on bone, in addition to its recognized suppressive effect on bone resorption. PMID- 11434667 TI - Expression of somatostatin receptors on human pituitary adenomas in vivo and ex vivo. AB - The distribution and biologic activity of somatostatin receptor subtypes (SSTR) in pituitary adenomas is not clarified, especially regarding clinically non functioning adenomas (NFPA). We therefore characterized SSTR in human pituitary adenomas by combining molecular biology and in vivo scintigraphy. Co-expression of gonadotropin-releasing hormone receptor (GnRH-R) mRNA was also assessed to see whether this feature was associated with adenoma subtype and SSTR status. Pituitary tumor biopsies were obtained during transsphenoidal adenomectomy from 21 patients (11 NFPA, 7 acromegalics, 2 prolactinomas, 1 Cushing's disease). Expression of mRNA encoding the 5 known SSTR subtypes and the GnRH-R was determined by RT-PCR. Twelve patients also underwent a pre-operative somatostatin receptor scintigraphy. Most adenomas (no.=18) expressed mRNA for more than one SSTR. SSTR2 mRNA was expressed in 18 cases, whereas SSTR4 was absent in all but one. SSTR3 was frequently expressed in NFPAs. Somatostatin receptor scintigraphy was positive in most cases, and with a significantly higher uptake index in GH producing adenomas all of which expressed SSTR2 mRNA. The uptake index appeared to be related to receptor density rather than tumor volume. Expression of GnRH-R mRNA was found in both NFPAs and GH-producing adenomas and was not significantly associated with a particular SSTR subtype population. IN CONCLUSION: 1) the distribution of SSTR is not significantly different between NFPA and GH-producing adenomas; and 2) somatostatin receptor scintigraphy reveals a higher uptake in GH producing adenomas which is not significantly related to either SSTR distribution or tumor volume. PMID- 11434668 TI - Increased visceral adipose tissue is associated with increased circulating insulin and decreased sex hormone binding globulin levels in massively obese adolescent girls. AB - The current study was designed to examine the relationship between body fat distribution, as evaluated by anthropometry and magnetic resonance imaging (MRI), and circulating insulin, sex hormone and SHBG levels in obese adolescent girls. Twenty-nine obese adolescent girls, aged 12.6-16.9 years with a mean BMI of 30.51+/-1.86 participated in this study. All girls had breast stage B4-5 and pubic hair stage P4-5. Percent obesity and BMI as indices of being overweight were calculated; the waist-to-hip ratio (WHR) and the waist-to-thigh ratio (WTR) were calculated to obtain two anthropometric indices for the pattern of body fat distribution. The areas of visceral (VAT) and subcutaneous adipose tissue (SAT) were evaluated by MRI at the L4-L5 level. Serum concentrations of total T, DHEAS, 17beta-estradiol, progesterone and SHBG were measured. Plasma glucose and insulin concentrations were evaluated during an oral glucose tolerance test. WHR was the only anthropometric parameter that was significantly associated with the area of VAT. Insulin level showed correlation with both WHR and the area of VAT; no correlation was found between insulin levels and WTR. Both WHR and VAT were negatively correlated with serum DHEAS level and positively correlated with T level. There were strong negative correlations between serum SHBG level and the area of VAT and WHR. Inverse correlation was found between serum SHBG level and insulin. Serum 17beta-estradiol and progesterone levels showed no significant correlation with all the patterns of body fat distribution. SAT was not significantly correlated with both anthropometric parameters and any of the sex hormones evaluated. We can draw two main conclusions. Firstly, in massively obese adolescent girls, the WHR seems to be a good indicator for the accumulation of VAT, and abdominal obesity, rather than adiposity per se, appears to be related to biochemical complications. Secondly, increased upper body adiposity and, in particular, the intra-abdominal fat area are associated with increased insulin levels in massively obese adolescent girls. The associated reductions in SHBG and DHEAS levels represent an early general risk factor for the development of metabolic and cardiovascular diseases in this population, as previously described for obese adult women. PMID- 11434669 TI - Post-surgical ablation of thyroid residues with radioiodine in Ukrainian children and adolescents affected by post-Chernobyl differentiated thyroid cancer. AB - Post-surgical ablation of thyroid residues with 131-iodine (131-I) is usually recommended after near-total thyroidectomy in high-risk patients, including children, with differentiated thyroid cancer (DTC). We report here the results of post-surgical radioiodine thyroid ablation in 249 children and adolescents of Ukraine with post-Chernobyl DTC initially treated with near-total thyroidectomy at the Institute of Endocrinology and Metabolism in Kiev, during a 2-year period. The patients' age at the time of the Chernobyl accident (1986), ranged from <1 to 14 yr in 223 subjects (children) and from 15 to 18 yr in 26 subjects (adolescents). Six weeks after surgery a diagnostic 131-I whole body scan revealed the presence of residual thyroid tissue in all cases. All patients received one or more courses of radioiodine therapy, for a total of 468 courses. One hundred and twenty-nine out of 249 patients (51.8%) were successfully ablated. The total number of treatment courses needed in these patients was 219. Most patients required multiple doses of radioiodine, only 63 required a single dose. One hundred and twenty patients (48.2%) treated with radioiodine were not ablated and are still under treatment program. The clinical features and the amount of thyroid residue were not different in ablated or not-ablated patients. Our results indicate that in this particular population of post-Chernobyl thyroid carcinomas, thyroid ablation is a rather difficult task. Only 51.8% were successfully ablated. Possible explanation for this finding may be the young age of the patients, other particular features of post-Chernobyl thyroid carcinoma or technical aspects, such as less radical surgical procedures. PMID- 11434670 TI - Multiple intracranial recurrent tumors with hyperprolactinemia combined with a parasellar malignant fibrous histiocytoma long after transfrontal surgery and irradiation to a pituitary adenoma. AB - We herein describe a 40-year-old woman with hyperprolactinemia, an empty sella and two extrasellar intracranial recurrent tumors which were revealed 23 years after the first transfrontal craniotomy and 18 years after the second transfrontal surgery and irradiation to a provable prolactin-producing pituitary macroadenoma. One recurrent tumor was in the right orbital apex causing right oculomotor nerve palsy, and the other tumor was in the right apex partispetrosae and foramen jugulare. Although her serum prolactin level decreased after the administration of bromocriptine mesilate, and the size of the two tumors remained unchanged, a malignant fibrous histiocytoma, which might have been induced by the irradiation 18 years before, grew rapidly in the right suprasellar-prepontine cistern to the right pedunculus cerebralis, leading to a poor prognosis. This case confirmed the importance of the life-lasting follow-up of pituitary adenomas treated with surgery and/or irradiation therapy. Not only ectopic recurrence of the primary tumor but also post-irradiation tumors may become apparent long after the removal of the primary tumor. PMID- 11434671 TI - Pituitary prolactin-secreting macroadenoma combined with bilateral breast cancer in a 45-year-old male. AB - We describe an unusual case of bilateral breast cancer synchronous with pituitary macroprolactinoma in a young male. Up to date, only very few of such cases have been described worldwide and to our knowledge this is the first one in which both breast cancer and pituitary macroadenoma have been found together at the time of presentation. A 45-year-old male was diagnosed as having a pituitary macroprolactinoma and bilateral breast cancer on the basis of hypogonadism (testosterone 2.9 pmol/l) with very high levels of prolactin (33,100 U/l), typical neuroradiologic finding of a pituitary macroadenoma, marked bilateral gynecomastia with mammographic pattern highly suspected for cancer and subsequent hystological confirmation. Bilateral mastectomy was performed and medical therapy with bromocriptine 10 mg/day was started. After 2-year follow-up the patient is disease-free. Hormonal, neuroradiological and oncological patterns are all negative or markedly improved. We stress the importance of prolactin for its possible biological effects on breast cancer induction or growth. Moreover in any case of hyperprolactinemia we suggest a mammographic examination and, in the case of breast cancer, at least a baseline hormonal profile. PMID- 11434673 TI - Evolving therapeutic strategies for acromegaly. PMID- 11434672 TI - The role of protein kinase C isoforms in insulin action. AB - Insulin action on target tissues is mediated by specific tyrosine kinase receptors. Upon ligand binding insulin receptors autophosphorylate and phosphorylate intracellular substrates on tyrosine residues. These early events of insulin action are followed by the activation of a number of enzymes, including protein kinase C (PKC). At least 14 PKC isoforms have been identified and cloned to date. PKCs appear to play dual roles in insulin signaling. For instance, they are involved in transduction of specific insulin signals but also contribute to the generation of insulin resistance. In this article, we will analyze the experimental evidence addressing the mechanism by which insulin might activate individual PKC isoforms as well as the role of single PKCs in insulin induced bioeffects. PMID- 11434674 TI - Endocrinology and art. Dwarf with hypergonadism. PMID- 11434675 TI - Post-prandial decrease of circulating human ghrelin levels. AB - Ghrelin, an endogenous ligand of the GH secretagogue-receptor, has recently been shown to stimulate GH secretion and to have orexigenic and adipogenic effects in rodents, but little is known about its regulation and biological function in humans. Gastric motor function is under control of the central nervous system; however, the afferent and efferent loops of this feedback control mechanism remain to be elucidated. In the study presented here we investigated the effect of nutrient intake on circulating human ghrelin levels, and a possible association between ghrelin levels and gastric emptying. Ten healthy volunteers received a standard meal after an overnight fast. Food intake significantly decreased plasma ghrelin levels from 248.5 +/- 15.0 to 179.5 +/- 17.9 fmol/ml (120 min after meal, p=0.047). Gastric emptying half-time (non-invasive 13C octanoic acid breath test) was correlated with fasting plasma ghrelin levels (r=0.74, p=0.0013). Ghrelin appears to be one possible candidate to provide feedback signaling between nutrient intake, gastric motor function and the central nervous system. PMID- 11434676 TI - Homozygous and compound-heterozygous type I plasminogen deficiency is a common cause of ligneous conjunctivitis. AB - Severe type I plasminogen deficiency has been recently linked to ligneous conjunctivitis, a rare and uncommon form of chronic conjunctivitis. In this study, eight unrelated ligneous conjunctivitis patients living in different parts of the world were examined. All affected subjects from which plasma was available displayed absent or markedly reduced plasminogen antigen and plasminogen functional activity. Molecular genetic studies of seven patients identified a Lys19-->Glu mutation in two boys in a homozygous state, and in two girls in a compound-heterozygous state in which the second plasminogen gene carried a missense (Arg134-->Lys) and a nonsense mutation (Cys133--> Stop), respectively. A fifth patient was shown to be homozygous for a frameshift mutation in plasminogen exon 14 (Gly565ins-G). In two unrelated subjects with ligneous conjunctivitis no mutations in the plasminogen gene were identified. Our results suggest that the Lys19-->Glu mutation is the most prevalent mutation in the plasminogen gene of patients with ligneous conjunctivitis. PMID- 11434677 TI - Factor X levels, polymorphisms in the promoter region of factor X, and the risk of venous thrombosis. AB - Elevated levels of procoagulant proteins factor II, factor VIII, factor IX, factor XI and fibrinogen are associated with an increased risk of venous thrombosis. In a population-based case-control study on venous thrombosis (Leiden Thrombophilia Study, LETS) we investigated whether elevated coagulation factor X (FX) levels are a risk factor for venous thrombosis and whether FX levels are determined by polymorphisms in the promoter region of the FX gene. We found that subjects with high FX levels (above the 90th percentile, > or = 126 U/dl) had a 1.6-fold increased risk of venous thrombosis. The highest risk (OR = 4.3, 95% confidence interval: 1.5-12) was found in the subgroup of premenopausal women who are not using oral contraceptives. However, these estimated risks disappeared after adjustment for other vitamin K-dependent coagulation factors II, VII and IX. To study the influence of genotypic variation on plasma FX levels we assessed four polymorphisms in the promoter region of the FX gene: a TTGTGA insertion between position -343A and -342G, a C/T polymorphism at position -222, a C/A polymorphism at position -220 and a C/T polymorphism at position -40. No relationship between these investigated genotypes and FX levels was observed. We conclude that high FX levels predict risk of thrombosis, but are not a risk factor for venous thrombosis when the levels of other vitamin K-dependent proteins are taken into account. PMID- 11434678 TI - Recruitment of labelled monocytes by experimental venous thrombi. AB - OBJECTIVE: Infusing monocytes that have been stimulated to produce fibrinolytic activators and factors that regulate cell proliferation, migration and maturation, might enhance venous thrombus resolution. The aim of this study was to determine the time course of infused monocyte recruitment into venous thrombus in an appropriate model of this disease. DESIGN AND METHODS: Thrombus was induced in the inferior vena cava of male Wistar rats using reduced flow (80-90% stenosis). The vessel wall was examined at 1hr by scanning electron microscopy. Resolving thrombi with surrounding vena cava were obtained at 1, 7, 14 and 21 days after induction (n = 8). Sections, taken at 0.5 mm intervals (10-15 sections per thrombus), were stained using haematoxylin, Martius Scarlet Blue and antibodies against monocytes, platelets and fibrin. Sections from human venous thrombi (n = 4) were similarly stained. The area occupied by monocytes (in relative pixel units, RPU) was determined using computer aided image analysis. Peripheral rat blood monocytes were extracted, fluorescently labelled and injected intravenously into 7 rats prior to thrombus induction, Vena cava with thrombus was harvested 1 h, 2, 3, 4, 7, 14 and 25 days after induction and their fluorescence measured. The fluorescent content of the caval wall and thrombus was analysed in greater detail at 2 and 25 days after thrombus induction (n = 4 at each time interval). RESULTS: Experimental thrombi were structurally similar to human thrombus and resolved within 14-21 days. Scanning electron microscopy showed minimal endothelial damage at 1 h with signs of early thrombus formation (platelet, red cell leukocyte and fibrin deposition). Neutrophils were the predominant leukocyte in the thrombus at 1 day, with monocytes making up only 0.3% (0.04% sem) of the area of the thrombus. There was a steady increase in thrombus monocyte content and by 21 days the percentage area of thrombus covered by monocytes had increased by over 35 fold to 11.5% (2.3% sem) (p <0.001). Initially, monocytes appeared around the edge of the thrombus and became more evenly distributed through the thrombus as resolution progressed. Labelled monocytes could be found in the circulation up to 1 week after infusion. The fluorescent content (RPU) of the thrombus increased over 25 days (mean RPU At 2 days 0.012, sem 0.005; mean RPU at 25 days 1.062, sem 0.252, p = 0.008). The number of labelled monocytes in the vessel wall peaked at 2 days and decreased thereafter. CONCLUSION: The structure of thrombi produced by this model was comparable to that of human venous thrombi. Endogenous and injected monocytes migrated into the thrombus during natural resolution, possibly via the vein wall. Monocyte targeting could therefore be used to develop novel treatments for venous thrombosis, with the aim of reducing post-thrombotic complications. PMID- 11434679 TI - Increased hepatic fibrinogen Bbeta-gene transcription is not enough to increase plasma fibrinogen levels. A transgenic mouse study. AB - The fibrinogen Aalpha, Bbeta, and gamma polypeptides are encoded by three separate genes, which are arranged in the order gamma-alpha-beta. In order to study the biosynthesis of fibrinogen in vivo we generated a line of transgenic mice carrying extra copies of the fibrinogen beta-gene. To clone the mouse fibrinogen Bbeta-chain gene, a mouse 129 Sv/Ev genomic cosmid library was screened, using the mouse fibrinogen Aalpha-, Bbeta-chain cDNA. A clone containing the complete fibrinogen Bbeta-chain gene including approximately 11-kb of the natural promoter region was identified and subsequently microinjected into mice. Southern blot analysis identified a founder that carried additional copies of the fibrinogen Bbeta-chain gene. Transgenic offspring of this founder were interbred and heterozygous and homozygous transgenic mice were obtained. Northern blot analysis demonstrated approximately a 3-fold increase in fibrinogen Bbeta mRNA in heterozygous mice as compared to wild-type, whereas homozygous transgenic mice showed approximately a 9-fold increase. The levels of the Aalpha and gamma mRNAs in transgenic homozygous mice were not changed as compared to those in wild type mice. Fibrinogen levels in plasma were not significantly increased in transgenic mice as compared to wild-type mice. These results indicate that: additional copies of the fibrinogen Bbeta-chain gene lead to increased levels of the Bbeta-chain mRNA in the liver; the increased levels of Bbeta-chain mRNA in homozygous overexpression mice do not change the transcription levels of the two other fibrinogen mRNAs in vivo; the absence of an increased plasma fibrinogen level in the transgenic mice indicates that this level is not regulated solely by transcription of the Bbeta-chain gene. PMID- 11434680 TI - Signal transduction pathways underlying the expression of tissue factor and thrombomodulin in promyelocytic cells induced to differentiate by retinoid acid and dibutyryl cAMP. AB - Acute promyelocytic leukaemia (APL) may be associated with disseminated intravascular coagulation, as a result of increased tissue factor (TF) expression and reduced thrombomodulin (TM) expression by APL blast cells. During retinoid acid (RA)- and dibutyryl cAMP (dbcAMP)-induced differentiation of the APL cells, there is a marked up-modulation of both the protein kinase A (PKA) and C (PKC) activities. In order to further assess whether these kinases are intimately associated with both the differentiation process and the regulation of TF and TM expression, we have correlated the modulation of their respective pathways with the extent of differentiation and modulation of these cellular receptors. NB4 cells were incubated with all-trans-RA (ATRA) or dbcAMP for up to 48 h. The contribution of phospholipase C (PLC), inositol phosphate (IP), PKC and PKA in the expression of CD11b, TF and TM was studied by the use of specific inhibitors. Myo-inositol uptake and PKC activity increased in cells induced to differentiate by ATRA but the retinoid did not affect cAMP levels or PKA activity. Under treatment with dbcAMP, PKA activity was increased while inositol uptake and PKC activity remained unchanged. Our results show that the effects of ATRA and dbcAMP on promyelocytic cells are closely related, respectively, to the PLC/IP/PKC and the cAMP/PKA pathways. In cells induced to differentiate by ATRA, CD11b expression seems more closely related to inositol uptake than to PKC activity while the expression of TF and TM show the opposite pattern, which suggests cellular events regulated at a different level within a common signal transduction pathway. PMID- 11434681 TI - Ectopic alphaIIbbeta3 integrin signaling involves 12-lipoxygenase- and PKC mediated serine phosphorylation events in melanoma cells. AB - Megakaryocytic genes such as alphaIIbbeta3 can be expressed by malignant cells as part of the disturbances in their gene regulation. However, the function of the gene product greatly depends on the interaction of the ectopic protein with the new environment. The outside-in signaling of the ectopically expressed alphaIIbbeta3 integrin was studied in B16a murine melanoma cells using a monoclonal antibody, specifically directed to the activated conformation of alphaIIbbeta3, PAC-1 and the physiological ligand, fibrinogen. Ligation of alphaIIbbeta3 induced down-regulation of FAK but serine phosphorylation of three protein bands, 20/21, 85 and 140 kDa within 1-15 min. Flow cytometry indicated that the ligation of the receptor in B16a cells induces approximately 50% increase in phosphoserine positive cells within 5-15 min. 12-lipoxygenase is placed downstream in the signaling pathway, since ligation of alphaIIbbeta3 induces 12-HETE production within 5 min and pretreatment of tumor cells with select lipoxygenase inhibitior, Baicalein, prevents the increase in serine phosphorylation. Confocal microscopy of adherent tumor cells demonstrated rearrangement of actin filaments upon alphaIIbbeta3 ligation paralleled by downregulation of p125FAK and phoshotyrosine+ adhesion plaques and translocation of PKCalpha to stress fibers and cortical actin. PKC appears to be the major effector serine kinase of the alphaIIbbeta3-coupled signaling pathway, since pretreatment of tumor cells with a select PKC inhibitor, Calphostin C, prevents the ligation-induced serine phosphorylation. Previous studies have indicated a role for the 12-lipoxygenase-PKC signaling pathway in platelet aggregation as well as tumor invasion, therefore the involvement of this cascade in the signaling of the ectopic alphaIIbbeta3 integrin may partially explain its role in tumor progression. PMID- 11434682 TI - The mRNAs of prekallikrein, factors XI and XII, and kininogen, components of the contact phase cascade are differentially expressed in multiple non-hepatic human tissues. AB - Recently RT-PCR studies had demonstrated the expression of plasma prekallikrein (PPK) mRNA in extrahepatic tissues. The questions arose whether that is illegitimate or regular expression, and whether the mRNAs of blood coagulation factors XI and XII, and high molecular weight kininogen, components of the contact activation cascade of blood coagulation are also expressed in non-hepatic tissues. These questions were addressed in the present study by employing quantitative RT-PCR. The relative mRNA levels of the respective proteins determined in 16 human tissues indicate legitimate extrahepatic transcription of at least three of the genes. Transcription of all genes was highest in the liver, but only PPK mRNA was detected in all 16 tissues, especially high levels in pancreas, kidney, testis, spleen and prostate. We conclude from these results that PPK is synthesized in significant amounts in non-hepatic tissues and that this locally synthesized PPK may have special local functions. PMID- 11434683 TI - Fine mapping of inhibitory anti-factor V antibodies using factor V C2 domain mutants. Identification of two antigenic epitopes involved in phospholipid binding. AB - Hemorrhagic factor V inhibitors frequently bind to the second C-type (C2) domain of factor V and interfere with phospholipid binding. To define specific residues recognized by inhibitors from four patients (one bovine thrombin-induced and three spontaneous antibodies), epitope mapping was performed using recombinant human factor V lacking most of the B-type domain (FV des B) and alanine substituted mutants within the C2 domain (FV des B C2 mutants). FV des B C2 mutants located in the region between Lys2060 and Glu2069 were resistant to inhibition by three IgG preparations including the bovine thrombin-induced antibody in both prothrombinase and phospholipid-binding assays. In contrast, mutations at Lys2087 and Lys2092/Glu2096 were significantly resistant to inhibition by the fourth IgG preparation in both prothrombinase and phospholipid binding assays. These results confirm interference of phospholipid binding by hemorrhagic factor V inhibitors and support the role(s) of these residues in phospholipid binding. PMID- 11434684 TI - The role of factor XI in a dilute thromboplastin assay of extrinsic coagulation pathway. AB - Blood coagulation has been thought to be composed of both intrinsic and extrinsic pathways. Recent evidence strongly supports the critical role of the extrinsic pathway in the initiation of blood coagulation. This investigation established an assay that examines the role of FXI in the thromboplastin-initiated (extrinsic) coagulation based on this new concept. Plasma clotting times were measured at different concentrations of thromboplastin with activated FXII inhibited (FXIIa inhibited Diluted Thromboplastin Time, FXIIaiDTT). Only at low concentrations of thromboplastin was FXIIaiDTT of FXI-deficient plasma significantly prolonged than that of normal plasma. Depletion of FXI from normal plasma prolonged its FXIIaiDTT and replenishment of FXI shortened it. FXIIaiDTTs of both FVIII deficient and FIX-deficient plasma were remarkably prolonged, and addition of normal plasma dose-dependently shortened it. Furthermore, earlier alpha-thrombin inhibition was directly correlated with decreasing FXa generation. The amount of FXa production was: platelet-rich plasma > platelet-poor plasma > FXI-deficient plasma. Therefore, our findings from the FXIIaiDTT assays not only support the critical role of extrinsic pathway in blood coagulation initiation, but also demonstrate the importance of FXI as an amplifier of thrombin generation in thromboplastin-initiated coagulation. PMID- 11434685 TI - The role of factor XI in thrombin generation induced by low concentrations of tissue factor. AB - Thrombin generation has been studied in the plasma of severely factor XI deficient patients under conditions in which contact activation did not play a role. In platelet-rich as well as platelet-poor plasma, thrombin generation was dependent upon the presence of factor XI at tissue factor concentrations of between 1 and 20 pg/ml i.e. approximately 0.01 to 0.20% of the concentration normally present in the thromboplastin time determination. The requirement for factor XI is low; significant thrombin generation was seen at 1% factor XI; at 10%, thrombin formation was nearly normalised. A suspension of normal platelets in severely factor XI deficient plasma did not increase thrombin generation. This implies that there is no significant factor XI activity carried by normal platelets, although the presence of factor XI and factor XI inhibitors in platelets cannot be ruled out. PMID- 11434686 TI - Polymorphisms in the prothrombin gene and their association with plasma prothrombin levels. AB - To find genetic causes of high plasma prothrombin levels, an established prothrombotic risk factor. we searched for sequence variations in the prothrombin gene. We selected subjects with the 20210-GG genotype (since the 20210-A allele is already known to be associated with high levels) and elevated prothrombin levels (> or = 130 U/dl) from the Leiden Thrombophilia Study (LETS). No mutations were found in the 1 kb promoter region of the prothrombin gene in seven individuals with an isolated high prothrombin level. Comparison of the allelic frequencies of four different polymorphisms in the prothrombin gene in healthy volunteers and in the control subjects among the selected LETS individuals indicated a higher frequency of the 19911-G allele in the latter group (allele frequency 52 vs. 78%, respectively). Homozygous carriers of the 19911-G allele had 8 U/dl higher plasma prothrombin levels than 19911-AA carriers. This difference in prothrombin levels did not affect the thrombotic risk in 20210-GG carriers. In heterozygous 20210-A carriers the odds ratio increased from 1.6 (95% CI: 0.6-4.3) in subjects with 19911-A to 4.7 (1.6-14.0) in subjects with 19911-G on the other prothrombin allele. PMID- 11434687 TI - A collaborative study to establish the 6th International Standard for factor VIII concentrate. AB - A study was carried out to replace the 5th WHO International Standard (IS) for factor VIII concentrate, because of depletion of stocks. Two candidate concentrates (X and Y) were assayed as potential replacements against the 5th IS for FVIII concentrate, in a collaborative study involving 33 laboratories. Collaborators were asked to use the ISTH/SSC recommendations, including pre dilution of concentrates in FVIII deficient plasma in their assays. Several laboratories performed more than one assay method and altogether there were 21 sets of assays with the one-stage method, 6 with the two-stage method and 26 with the chromogenic method. There was good agreement between laboratories using each method for the comparison of concentrates X and Y against the 5th IS, but the overall potencies by one-stage and chromogenic methods each differed by approximately 5% from the overall mean, with the chromogenic potency approximately 10% higher than the one-stage. Inter-laboratory agreement was slightly better for concentrate Y than X, and stability studies indicated that Y was more stable than X. After considering all the information, together with comments from participants and from the FVIII/FIX Subcommittee of the ISTH/SSC, candidate Y (NIBSC code [97/616]), was proposed and accepted in October, 1998, by the Expert Committee on Biological Standardisation of the World Health Organisation to be the 6th International Standard for Factor VIII Concentrate with an assigned potency of 8.5 IU/ampoule. PMID- 11434688 TI - Adenovirus-mediated expression and packaging of tissue-type plasminogen activator in megakaryocytic cells. AB - Platelets release large quantities of plasminogen activator inhibitor 1 (PAI-1) that plays an important role in maintaining the integrity of fibrin-rich thrombi. In addition, tissue-type plasminogen activator (t-PA), a key physiological regulator of fibrinolysis, has been detected in platelet alpha-granules at low abundance. This information raises the possibility of enhancing t-PA expression in megakaryocytes as a means to enhance the fibrinolytic properties of platelet alpha-granules and target PAs directly to fibrin clots. This study was initiated to investigate adenovirus (Ad)-mediated expression and packaging of t-PA into alpha-granules-like structures in the megakaryocytic cell line MEG-01. Ad/t-PA infection of phorbol myristate acetate (PMA)-differentiated MEG-01 cells increased cellular t-PA levels by 120 fold (1580 +/- 130 ng/10(6) cells at 5 MOI) in comparison to non-or Ad/beta-gal-infected cells. Fluorescence-activated cell sorter (FACS) analysis indicates that Ad/t-PA-infected cells yielded a homogenous shift in the t-PA staining profile with a 4-fold shift in mean fluorescence in comparison to non- or Ad/beta-gal-infected cells. For the isolation of alpha granule-like structures, MEG-01 cell homogenates were fractionated by differential centrifugation and two consecutive Percoll density gradients. Fibrin autography of storage granules revealed a prominent lytic zone at Mr 66 kD comigrating with free t-PA. Quantitative analyses indicate that a 16-fold elevation in t-PA antigen within storage granules in comparison to non- or Ad/beta-gal-infected cells. To document the ability of t-PA to be stored in a rapidly-releasable form in these cells, we isolated platelet-like particles from the supernatant of differentiated cells and determined that particles from Ad/t PA-infected cells display a 4-8 fold enhanced secretion of t-PA following treatment with the clasical secretagogue calcium ionphore 23187, ADP, or thrombin. Confocal immunofluoresence microscopy analysis indicates that Ad/t-PA mediated productive expression of t-PA in murine megakaryocytes. These data provide support for the concept of increasing the expression of t-PA in megakaryocytes as a means to alter the hemostatic properties of alpha-granules. PMID- 11434689 TI - Regulation of PAI-1 concentration in platelets by systemic administration of antisense oligonucleotides to rats. AB - In this report we tested the effect of oligodeoxyribonucleotides antisense to PAI 1 mRNA administered into rats on PAI-1 concentration in platelets. Low doses of the antisense oligonucleotide (MPO-16R) reduced PAI-1 activity, both in rat blood plasma and platelet lysates by 20.5% and 28.7%, respectively. There was no change in platelet count after treatment with MPO-16R but treated platelets showed lower aggregability as compared with controls (37 +/- 13% and 54 +/- 12%, respectively). In an experimental model of rat arterial thrombosis, low doses of MPO-16R caused a significant delay in the occlusion time (31.8%). These data further support for the role of PAI-1 as a major determinant of arterial thrombolysis resistance and for the first time demonstrate the possibility of reduction of platelet PAI-1 concentration by antisense approach. PMID- 11434690 TI - A new approach in the study of the molecular and cellular events implicated in heparin-induced thrombocytopenia. Formation of leukocyte-platelet aggregates. AB - Heparin-induced thrombocytopenia (HIT), a relatively common complication of heparin therapy, results of platelet activation, via the receptor for the Fc domain of IgG (FcgammaRIIa), by heparin-dependent-antibodies, commonly directed against the heparin-platelet factor 4 (H-PF4) antigenic complex. Our strategy was to use whole blood allowing the study of leukocyte-platelet interactions. Experiments were performed with blood from healthy donors incubated with HIT patients' plasma and different concentrations of heparin. We showed that 75% of the HIT patients' plasma induced the formation of leukocyte-platelet-aggregates in a heparin-dependent-manner. The formation of leukocyte-platelet-aggregates induced by HIT plasma in the presence of heparin was (i) independent of the healthy blood donor FcgammaRIIa polymorphism, (ii) correlated with the levels of anti H-PF4 IgG antibodies contained in the patients' plasma, and to a lesser extent to anti H-PF4 IgM antibodies, and (iii) was mediated by P-selectin. This report opens new prospects in the study of the molecular and cellular events implicated in HIT. PMID- 11434691 TI - The effect of platelet PlA polymorphism on experimental thrombus formation in man depends on blood flow and thrombogenic substrate. AB - A number of studies have reported conflicting data on the association of the PlA1/PlA2 polymorphism of the GPIIIa gene and coronary syndromes. We have investigated the effect of this polymorphism on experimental platelet thrombus formation in man. Forty healthy male volunteers were genotyped for the PlA1/PlA2 polymorphism. Thrombus formation was induced ex vivo by exposing a tissue factor (TF) or a collagen-coated coverslip in a parallel plate perfusion chamber to native blood for 2 and 4 min. The shear rates at these surfaces were 650 and 2,600 s(-1). Platelet and fibrin deposition was quantified by immunoenzymatic methods. The frequencies of PlA1/PlA1 and PlA1/PlA2 genotypes were 52.5% and 47.5%, respectively. Ex vivo deposition of fibrin on TF was not affected by the PlA1/PlA2 polymorphism. However, the ex vivo platelet deposition at 650 s(-1) was higher in blood from PlA1/PlA1 individuals than in PlA1/PlA2 individuals (P= 0.008 at 4 min). On collagen, neither fibrin nor platelet deposition was significantly affected by the PlA1/PlA2 polymorphism. Platelet thrombus formation is significantly influenced by genetic variations in the GPIIIa platelet receptor. This effect depends on the blood flow properties and the nature of the thrombogenic stimulus. PMID- 11434692 TI - Thrombin-induced MCP-1 expression involves activation of the p22phox-containing NADPH oxidase in human vascular smooth muscle cells. AB - Activation of vascular smooth muscle cells (VMSC) by thrombin induces the expression of the chemokine, monocyte chemoattractant protein-1 (MCP-1). We investigated in cultured human and rat VSMC whether reactive oxygen species (ROS) derived from the vascular NADPH oxidase contribute to this effect. Exposure of cultured VSMC to thrombin rapidly increased ROS formation, phosphorylation of p38 MAP kinase as well as the expression of MCP-1. Specific inhibition of the p22phox subunit of the vascular NADPH oxidase using either p22phox neutralizing antibody or p22phox antisense oligonucleotides attenuated thrombin-induced ROS generation. Furthermore, thrombin-induced p38 MAP kinase activation as well as MCP-1 expression were impaired by antioxidants as well as by p22phox antisense oligonucleotides. Inhibition of p38 MAP kinase diminished the thrombin-induced expression of MCP-1. CONCLUSION: Thrombin, by activating a p22phox-containing NADPH oxidase, elicits ROS generation and activation of p38 MAP kinase in VSMC. The subsequent induction of MCP-1 expression highligts the crucial role of the p22phox-containing NADPH oxidase in thrombin-induced signal transduction in VSMC. PMID- 11434693 TI - Adipose tissue expression of gelatinases in mouse models of obesity. AB - Following the observation by Brown et al. (Am J Physiol 1997; 272: C937-49) that primary rat adipocytes in culture secrete gelatinase A (MMP-2), we have evaluated gelatinase expression in adipose tissue with the use of mouse models of obesity. Wild-type mice were kept on a standard fat diet (SFD) or on a high fat diet (42% fat, HFD) and- genetically obese db/db mice were kept on SFD; gonadal and subcutaneous fat pads were removed and analysed ex vivo. These studies revealed that: 1) the HFD induced adipocyte hypertrophy; 2) after 32 weeks, significantly higher levels of 70 kDa (p <0.05) and 65 kDa proMMP-2 (p <0.01) were observed in extracts of gonadal fat pads of mice on HFD; 3) the contribution of active MMP-2 to the total level was comparable in SFD and HFD groups (20 to 30%); and 4) gelatinase B (MMP-9) was not consistently detected. These findings were confirmed by gelatin zymography and by mRNA determination using competitive RT-PCR. The presence of MMP-2 in the adipose tissue was confirmed immunologically and its localization in adipocytes revealed by immunogold electron microscopy. The potential functional role of MMP-2 in adipose tissue remains to be determined. PMID- 11434694 TI - Does thrombus contribute to in-stent restenosis in the porcine coronary stent model? PMID- 11434695 TI - Protein C concentrates in a neonate with a cerebral venous thrombosis due to heterozygous type 1 protein C deficiency. PMID- 11434696 TI - Vascular endothelial growth factor in plasma of patients undergoing peripheral angioplasty. PMID- 11434697 TI - High prevalence of FVHR2 polymorphism in Costarician Indians who have no FVL. PMID- 11434698 TI - A hypothesis to explain the reported association of the alpha-fibrinogen A312 allele with thromboembolic disease. PMID- 11434699 TI - Identification of novel polymorphisms within the protein Z gene, haplotype distribution and linkage analysis. PMID- 11434700 TI - Heparin-induced thrombocytopenia: yet another treatment paradox? PMID- 11434701 TI - A comparison of danaparoid and lepirudin in heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is a hypercoagulable syndrome strongly associated with thrombosis that is usually treated with drugs that inhibit factor Xa (danaparoid) or thrombin (lepirudin). In the present study the outcome of HIT patients treated with danaparoid or lepirudin was compared using the single or combined endpoints of new thromboembolic complications (new TECs), amputations and/or death, and major bleeding. HIT-patients treated with lepirudin were enrolled in two prospective trials and patients, who were identified in the same two laboratories during the same time period, who were not enrolled into these studies but treated with danaparoid, were assessed retrospectively according to a standardized questionnaire. 126 danaparoid (60.3% female) and 175 lepirudin treated patients (58.3% female) fulfilled the same inclusion and exclusion criteria. In a time-to-event-analysis the cumulative risk of combined endpoint was higher in HIT-patients without thromboembolic complication at baseline treated with danaparoid (usually in prophylactic dose 750 anti-factor Xa units b.i.d. or t.i.d.s.c.) as compared to lepirudin (aPTT adjusted) (P = 0.020). Whereas HIT-patients with TEC at baseline who were usually treated with therapeutic dose had a similar outcome in both treatment groups (P = 0.913). Major bleeding occurred in 2.5% (95% CI 0.5-7.0%) of danaparoid treated patients as compared to 10.4% (95% CI 6.3-15.9%) of lepirudin treated patients until day 42 (P = 0.009). This indicates that the efficacies of therapeutic doses of danaparoid or lepirudin in preventing death, amputation or new TEC in HIT patients do not differ largely, but the risk of bleeding seems to be higher in lepirudin treated patients. The prophylactic dose of danaparoid approved in the European Union for HIT without TEC at baseline seems suboptimal. A prospective comparative trial is required to verify these preliminary conclusions. PMID- 11434702 TI - A cell-based model of hemostasis. AB - Based on our work and that of many other workers, we have developed a model of coagulation in vivo. Many workers have demonstrated mechanisms by which cells can influence the coagulation process. Nonetheless, the prevailing view of hemostasis remains that the protein coagulation factors direct and control the process with cells serving primarily to provide a phosphatidylserine containing surface on which the procoagulant complexes are assembled. By contrast, we propose a model in which coagulation is regulated by properties of cell surfaces. This model emphasizes the importance of specific cellular receptors for the coagulation proteins. Thus, cells with similar phosphatidylserine content can play very different roles in hemostasis depending on their complement of surface receptors. We propose that coagulation occurs not as a "cascade", but in three overlapping stages: 1) initiation, which occurs on a tissue factor bearing cell; 2) amplification, in which platelets and cofactors are activated to set the stage for large scale thrombin generation; and 3) propagation, in which large amounts of thrombin are generated on the platelet surface. This cell based model explains some aspects of hemostasis that a protein-centric model does not. PMID- 11434703 TI - Reduction of pain episodes and prothrombotic activity in sickle cell disease by dietary n-3 fatty acids. AB - The effects of dietary n-3 fatty acids (n-3FAs) on the frequency of pain episodes and ex vivo blood tests of thrombosis have been evaluated in patients with sickle cell disease (SCD) utilizing a double-blind, olive oil-controlled clinical trial. Dietary n-3FA therapy (0.1 g/kg/d) was provided as menhaden fish oil (0.25 g/kg/d) containing 12% eicosapentaenoic acid (EPA), and 18% docosahexaenoic acid (DHA). Within 1 month dietary n-3FAs exchanged with n-6FAs in plasma and erythrocyte membrane phospholipids (p <0.01 in all cases). Treatment with dietary n-3FAs for 1 year reduced the frequency of pain episodes requiring presentation to the hospital from 7.8 events during the preceding year to 3.8 events/year (p <0.01; n = 5). By contrast, subjects receiving control dietary olive oil (n = 5) experienced 7.1 pain events/year, compared to 7.6 during the previous year (p >0.4). The reduction in episodes in n-3FA-treated subjects was also significant when compared to control subjects (p <0.01). Dietary n-3FA therapy was not associated with hemorrhagic, gastrointestinal or other adverse effects. Compared to 10 asymptomatic African-American controls, sickle cell subjects demonstrated significantly increased pretreatment: 1) flow cytometric expression of platelet membrane P-selectin (CD62p; p <0.01) and annexin V binding sites (p = 0.02); 2) plasma levels of platelet-specific secretory proteins platelet factor 4 (PF4) and beta-thromboglobulin (betaTG) (p <0.01 in both cases); 3) plasma products of thrombin generation, prothrombin fragment 1.2 (F1.2) and thrombin:antithrombin (TAT) complex (p <0.01 in both cases); and 4) plasma levels of thrombolytic products, D-dimer and plasmin:antiplasmin (PAP) complex (p <0.01 in both cases). Treatment with dietary n-3FAs concurrently decreased plasma levels of F1.2, D dimer, and PAP (p <0.05, compared to olive oil controls), implying that the reduction in pain events was related to n-3FA-dependent inhibition of thrombosis. We conclude that dietary n-3FAs reduce the frequency of pain episodes perhaps by reducing prothrombotic activity in sickle cell disease. PMID- 11434704 TI - von Willebrand factor-cleaving protease in childhood diarrhoea-associated haemolytic uraemic syndrome. AB - A deficiency of von Willebrand factor (vWF)-cleaving protease, either due to a congenital deficiency or to the presence of a protease inhibitor of vWF-cleaving protease has been associated with thrombotic thrombocytopenic purpura (TTP). We have studied vWF-cleaving protease in diarrhoea-associated haemolytic uraemic syndrome (D+ HUS), which shares clinical features with TTP. 29 children with acute D+ HUS and 13 control children were studied. vWF-cleaving protease activity was normal (range 50-150%) in 39 of 42 plasma samples. Levels of protease activity between 25 and 50% were noted in plasma from two D+ HUS patients. One D+HUS patient, who had clinical features of TTP, had a vWF-cleaving protease inhibitor producing a severe deficiency of vWF-cleaving protease. Thus a deficiency of vWF-cleaving protease appears to be atypical in D+HUS. The detection of a vWF-cleaving protease inhibitor in one patient suggests it may be associated with infection such as E. coli O157. PMID- 11434705 TI - The effect of unfractionated vs. low molecular weight heparin on tissue factor pathway inhibitor levels in hospital inpatients. AB - Although heparin is widely used as an antithrombotic agent, its multiple mechanisms of action are not fully defined. Recent work has suggested that tissue factor pathway inhibitor (TFPI) may contribute to the antithrombotic activity of heparin by inhibiting the extrinsic pathway of coagulation. We have investigated the effect of heparin on TFPI and have found that when unfractionated heparin is given by continuous intravenous infusion to hospitalized inpatients, TFPI levels increase 2.3-fold and remain high as long as heparin is continued, but return to baseline levels soon after the infusion is stopped. In contrast, therapeutic doses of the low molecular weight heparin, dalteparin, resulted in significantly less TFPI induction. Given the increasing number of studies establishing the clinical efficacy of low molecular weight heparins as antithrombotic agents, these results suggest that TFPI may not be a major contributor to the antithrombotic effect of heparin. PMID- 11434706 TI - High prevalence of the I278T mutation of the human cystathionine beta-synthase detected by a novel screening application. AB - Classical homocystinuria due to cystathionine beta-synthase deficiency is one of the disorders revealing a high risk of thromboembolic events and vascular disease. This autosomal-recessively inherited metabolic disorder is considered to be rare with an estimated prevalence of 1:130,000 in the German population. In this study, we developed a novel multiplex PCR generating allele specific fragment lengths to analyse individual genotypes of the two most frequent cystathionine beta-synthase alterations, the I278T mutation, which is worldwide found on up to the half of homocystinuric alleles, and the adjacent polymorphism 844ins68. Screening of 200 unrelated German control subjects revealed a frequency of heterozygosity of 1.5% for 1278T corresponding to a calculated frequency of homozygosity of 1:17.800. Our data indicate that homocystinuria due to cystathionine beta-synthase deficiency is a frequently unrecognized disorder resulting in a high risk of thromboembolic events. PMID- 11434707 TI - Platelet reactivity in acute coronary syndromes: evidence for differences in platelet behaviour between unstable angina and myocardial infarction. AB - Previous work has shown that P-selectin and mean platelet volume, two markers associated with platelet reactivity, are elevated in acute coronary syndromes. This study investigated the possibility that these markers may define unstable angina (UA) and acute myocardial infarction (MI) as two separate conditions based on platelet behaviour. Mean platelet volume (MPV) was higher in UA patients (n = 15) than in those diagnosed with MI (n = 15) (10.7 +/- 0.25 fL, vs. 9.8 +/- 0.27 fL, P = 0.005). Platelet count was lower in UA than in MI (215 +/- 13 x 10(9)/L vs. 271 +/- 20 x 10(9)/L, P = 0.03). The percentage of platelets expressing P selectin was higher in MI than in UA (9.1 +/- 1.9% vs. 4.2 +/- 0.85%, P = 0.03). This parameter was positively correlated with MPV in UA (r = 0.5, P = 0.04) but negatively correlated in MI (r = -0.6, P = 0.01), with no correlation for ACS as a whole (r = -0.32, P = 0.1). Our results suggest that in MI there is an acute process of generalised platelet activation that is unrelated to changes in MPV, whereas in UA there is an ongoing process of platelet consumption that leads to an increase in platelet size to compensate for a persistent decrease in platelet count. This study suggests that there is a fundamental difference in platelet biology between these two diseases. PMID- 11434708 TI - Diagnostic accuracy of triage tests to exclude pulmonary embolism. AB - We performed a study in 403 prospectively included patients with suspected pulmonary embolism to compare the accuracy of a combination of the SimpliRED D dimer assay and an intuitive clinical probability estimate with either one alone. Based on a conjoint diagnostic refer, ence standard, including ventilation perfusion lung scintigraphy and pulmonary angiography, the prevalence of pulmonary embolism was 31%. We demonstrated a high sensitivity (98%, 95% CI 95 100) and negative predictive value (94%, 95% CI 79-99) for the combination of the two tests. These figures were more favorable than for either test alone. The specificity of the combination was lower (11%, 95% CI 9-12) and consequently the proportion of patients in whom further diagnostic tests would have been avoided was only 8%. We conclude that the combined use of the SimpliRED test and the clinical probability estimate attains a higher sensitivity than either test alone. However, there remains a risk of false negatives and the exclusion efficiency is limited. PMID- 11434709 TI - Plasma levels of free and total TFPI, relationship with cardiovascular risk factors and endothelial cell markers. AB - Free-TFPI (f-TFPI) presents high anticoagulant activity and its plasma level correlates with unfavorable outcomes in unstable angina. Total TFPI (t-TFPI) represents mainly the lipid-bound form which seems to have a poor anticoagulant activity. Until now, it is not known whether the variations of f-TFPI plasma levels are determined by environmental factors. The aim of our study was to evaluate the influence of cardiovascular risk factors on plasma levels of f-TFPI and relations with other endothelial derived molecules in a population of 626 patients (277 men and 349 women) attending a metabolic ward for primary prevention of coronary disease. Free and total TFPI plasma levels were poorly correlated. f-TFPI levels increased with age in both sexes, t-TFPI in women only. Age-adjusted correlations of TFPI levels with conventional cardiovascular risk factors and endothelial cell markers showed different patterns for f-TFPI and t TFPI. f-TFPI correlated with parameters associated with insulin resistance, particularly in females. f-TFPI was also positively associated in both genders with fibrinogen and endothelial cell markers: t-PA, thrombomodulin (TM) and von Willebrand factor (vWF). t-TFPI correlated strongly with LDL-C in both sexes. It also correlated negatively with parameters of the insulin resistance syndrome. t TFPI also correlated with TM but not with other endothelial cell markers. The results of the multivariate step by step analysis showed that cardiovascular risk factors poorly explained the f-TFPI variability (7% and 4% in men and women, respectively), whereas they explained 16 and 20% of t-TFPI variability in men and women respectively (mostly related to LDL-C). In conclusion, this study showed that free- and total-TFPI are regulated differently. f-TFPI strongly correlates with endothelial cell markers and t-TFPI is more related to conventional cardiovascular risk factors. The strong gender effect on f-TFPI levels remains to be explained. PMID- 11434710 TI - HIPAA: wake-up call for health care providers. AB - The Health Insurance Portability and Accountability Act (HIPAA) may have an overall negative financial impact on health care providers across the board because of the significant amount of capital expenditures that could be required to ensure compliance (including, potentially, a complete overhaul of many organizations' technological, process, and security systems). The impact of HIPAA on an individual health care system will be determined by the current state of that organization's information technology system, as well as its preparedness in the coming months. Although it is extremely difficult to estimate total systemwide costs of HIPAA compliance, estimates range from $5.8 billion to $43 billion. PMID- 11434711 TI - Income, location, and the demand for health care from public, nonprofit, and for profit hospitals. AB - Although, empirically, for-profit hospitals serve few poor and indigent patients, they may be able to shift capital more quickly than hospitals of other ownership types, thereby spatially avoiding poor patients. However, in a market with a relatively high proportion of for-profit hospitals, spatial avoidance of poor patients is not possible because spatial competition will exist in non-poor areas. The study examines hospital choice for maternity care in a market with many for-profits using a gravity model or conditional logit. The analysis shows that poor and Medicaid populations choose for-profit hospitals overall. Income, along with distance, is an important factor in hospital choice. PMID- 11434712 TI - Impact of global bundled payments on hospital costs of coronary artery bypass grafting. AB - The Health Care Financing Administration began the Medicare Participating Heart Bypass Center Demonstration in 1991, in which hospitals and physicians are paid a single negotiated global price for all inpatient care for heart bypass patients. This article analyzed the changes in total and departmental direct variable costs during the 1991-1993 period using micro-cost data. The results indicate that all participating hospitals had significant reductions in total direct variable costs, after controlling for preoperative risk factors and postoperative outcomes. However, the patterns in cost reductions across major departments were different across hospitals. The cost reductions primarily came from nursing intensive care unit, routine nursing, pharmacy, and catheter lab. PMID- 11434713 TI - Physician manpower and health care expenditures in the United States: a thirty year perspective. AB - Data gathered on the number of physicians and health care costs in the United States over a 30-year period from 1970 through 1999 were correlated and analyzed using bivariate linear regression. The rise in the number of practicing physicians theoretically predicts 96 percent of the variability in rise of total U.S. health expenditures, 96 percent of the variability in rise of physician service expenditures, 98 percent of the variability in rise of hospital service expenditures, and 86 percent of the variability in rise of prescription pharmaceutical expenditures. If the rate of growth in the number of physicians continues unchanged in the current fee-for-service climate, the model herein predicts that the growth in health care costs will likely continue to rise substantially faster than the rate of growth of the U.S. population and the rate of inflation. The addition of nonphysician providers with the same clinical privileges will likely exacerbate the situation of rapidly escalating medical costs. It is also possible that the excess number of physicians and other health care providers with the same clinical privileges as physicians may decrease the overall quality of health care. Reducing the rise in the number of physicians and nonphysician practitioners may help to decrease overall health care expenditures and improve overall quality of health care in the United States. PMID- 11434714 TI - Diversion of prescription drugs to the black market: what the states are doing to curb the tide. AB - The diversion of legitimate controlled substances to the black market is a major cause of medical emergencies, fatalities, and drug-related dependencies. The effects harm not only the illegal user, but also the legitimate patient who may be getting shorted on treatments and innocent medical providers who may be charged with false claim offenses or other professional failures because of diversions that take place during their watch. The dollar magnitude of this crime is estimated to rival the black markets for both crack cocaine and heroine combined. This article addresses the various ways prescription drugs are diverted to the black market, some monitoring programs employed by the states, and guidelines that doctors, pharmacists, and other providers can use to protect themselves against possible liabilities arising from the diversion of prescription drugs. We will also address some of the oppositions to monitoring programs that have been asserted and replies to these oppositions. PMID- 11434715 TI - Final HIPAA privacy rules: "How do we get started?". AB - This article provides a general overview of the HIPAA privacy standards established by the Final Rules. It also discusses organizational approaches health care providers, more specifically hospitals and health systems, can use to "get started" with their HIPAA compliance effort. It also explores the corresponding human and financial resources that such providers will need to develop and implement the program. While the needs and approaches identified and discussed in this article may also apply to some extent to Covered Entities other than health care providers, this article does not specifically address those considerations in the context of health plans (including employer-sponsored plans), health care clearinghouses, or retail pharmacies. PMID- 11434716 TI - Physician services in an academic neurology department: using the resource-based relative-value scale to examine physician activities. AB - The inability of physician managers and decision makers to critically analyze the resource utilization of physicians has hindered a more comprehensive understanding of the role of neurologists in the patterns and organization of medical practice. This article outlines an approach for using the physician work relative value units (RVUs) in the Medicare Fee Schedule (MFS) to address this problem and profile physician clinical activities in a comparative manner. These techniques are then used to profile the physician services associated with the neurology department at a large academic hospital. All 28,048 physician services associated with a neurology department in 1995 were studied. Using billing data, physician work RVUs were assigned to each service and the results analyzed by major services, type of service, and physician workload for physician work RVUs and physician charges. For the average service, mean physician charges were $187 per service while median physician charges were $120. Mean physician work RVUs per service averaged 1.3 RVUs, and the median was 0.94 per service. Of all the services provided in the neurology department, 65 percent are visits and consultations, while medicine services (e.g., nerve conduction studies, needle electromyography, neuropsychological testing, and electroencephalogram) make up 31 percent. All the other services combined represented less than five percent of the services in the department. The top five physicians in the department account for 33 percent of all physician work RVUs in the neurology department. Using the physician work relative values in the MFS provides a unique perspective for analyzing and understanding neurologists' work activities. PMID- 11434717 TI - Toxic thyroid adenoma and toxic multinodular goiter. PMID- 11434718 TI - JMM, past and present. On dysmenorrhea due to mechanical causes, 1865. PMID- 11434719 TI - Mechanistic insights on the inhibition of tumor angiogenesis. AB - Angiogenesis, the growth of new vasculature, is an absolute requirement for the maintenance and progression of the overwhelming majority of the solid tumors. Unraveling the mechanisms that govern this complex biological process has become a central issue not only for understanding of the molecular basis of cancer but also for developing new therapeutic approaches that interfere with neovascularization of the tumor mass. Here we discuss the survival and apoptosis of endothelial cells in the context of vessel formation and regression in response to mediators of angiogenesis produced by tumors. It is the balance between proangiogenic and antiangiogenic molecules in the microenvironment of a vessel in vivo that determines whether the existing vasculature will expand, remain the same, or regress. Here we propose that the vascular endothelial cells themselves interpret and respond to these environmental cues by integrating the activities of the survival and apoptotic pathways within the cell. Thus it is the survival or death of the vulnerable cells that venture out to form new vessels that is the ultimate arbiter of whether neovascularization, as well as the growth of a malignancy that depends on it, succeeds or fails. PMID- 11434720 TI - Helper T cell anergy: from biochemistry to cancer pathophysiology and therapeutics. AB - Tolerance in vivo and its in vitro counterpart, anergy, are defined as the state in which helper T lymphocytes are alive but incapable of producing IL-2 and expanding in response to optimal antigenic stimulation. Anergy is induced when the T cell receptor (TCR) is engaged by antigen in the absence of costimulation or IL-2. This leads to unique intracellular signaling events that stand in contrast to those triggered by coligation of the TCR and costimulatory receptors. Specifically, anergy is characterized by lack of activation of lck, ZAP 70, Ras, ERK, JNK, AP-1, and NF-AT. In contrast, anergizing stimuli appear to activate the protein tyrosine kinase fyn, increase intracellular calcium levels, and activate Rap1. Moreover, anergizing TCR signals result in increased intracellular concentrations of the second messenger cAMP. This second messenger upregulates the cyclin-dependent kinase (cdk) inhibitor p27kip1, sequestering cyclin D2-cdk4, and cyclin E/cdk2 complexes and preventing progression of T cells through the G1 restriction point of the cell cycle. In contrast, costimulation through CD28 prevents p27kip1 accumulation by decreasing the levels of intracellular cAMP and promotes p27kip1 down-regulation due to direct degradation of the protein via the ubiquitin-proteasome pathway. Subsequent autocrine action of IL-2 leads to further degradation of p27kip1 and entry into S phase. Understanding the biochemical and molecular basis of T cell anergy will allow the development of new assays to evaluate the immune status of patients in a variety of clinical settings in which tolerance has an important role, including cancer, autoimmune diseases, and organ transplantation. Precise understanding of these biochemical and molecular events is necessary in order to develop novel treatment strategies against cancer. One of the mechanisms by which tumors down-regulate the immune system is through the anergizing inactivation of helper T lymphocytes, resulting in the absence of T cell help to tumor-specific CTLs. Although T-cells specific for tumor associated antigens are detected in cancer patients they often are unresponsive. Reversal of the defects that block the cell cycle progression is mandatory for clonal expansion of tumor specific T cells during the administration of tumor vaccines. Reversal of the anergic state of tumor specific T cells is also critical for the sufficient expansion of such T cells ex vivo for adoptive immunotherapy. On the other hand, understanding the molecular mechanisms of anergy will greatly improve our ability to design novel clinical therapeutic approaches to induce antigen-specific tolerance and prevent graft rejection and graft-versus-host disease. Such treatment approaches will allow transplantation of bone marrow and solid organs between individuals with increasing HLA disparity and therefore expand the donor pool, enable reduction in the need for nonspecific immunosuppression, minimize the toxicity of chemotherapy, and reduce the risk of opportunistic infections. PMID- 11434721 TI - Detection of thyroid-stimulating hormone receptor and Gsalpha mutations: in 75 toxic thyroid nodules by denaturing gradient gel electrophoresis. AB - The actual frequency of constitutively activating thyrotropin receptor or Gsalpha mutations in toxic thyroid nodules (TTNs) remains controversial as considerable variation in the prevalence of these mutations has been reported. We studied a series of 75 consecutive TTNs and performed mutation screening by the more sensitive method of denaturing gradient gel electrophoresis (DGGE) in addition to direct sequencing. Furthermore, the likelihood of somatic mutations occurring in genes other than that for the thyroid-stimulating hormone receptor (TSHR) and exons 7-9 of the Gsalpha protein gene was determined by clonality analysis of TTNs, which did not harbor mutations in the investigated genes. In 43 of 75 TTNs (57%) constitutively active TSHR mutations were identified. Six TSHR mutations were detected only by DGGE, underlining the importance of a sensitive screening method. Novel, constitutively activating mutations were identified at positions 425 (Ser-->Leu) and 512 (Leu-->Glu/Arg). Furthermore, a new base substitution was detected at position Pro639Ala (CCA-->GCA). Ten of 20 TSHR or Gsalpha mutation negative cases (50%) showed nonrandom X-chromosome inactivation, indicating clonal origin. In conclusion, somatic, constitutively activating TSHR mutations appear to be a major cause of TTNs (57%), while mutations in Gsalpha play a minor role (3%). The mutation negative but clonal cases indicate a probable involvement of somatic mutations other than in the TSH receptor or Gsalpha genes as the molecular cause of these hot nodules. PMID- 11434722 TI - Tumor regression induced by intratumoral injection of DNA coding for human interleukin 12 into melanoma metastases in gray horses. AB - Preclinical studies investigating new therapeutic principles against melanoma are presently being carried out in mouse models; however, these are not optimal. Here we describe a novel animal model using gray horses. These animals spontaneously develop metastatic melanoma that resembles human disease and is thus highly relevant for preclinical studies testing new immunotherapy protocols. We found that injection of plasmid DNA coding for the human cytokine interleukin 12 into established metastases induced significant regression in all 12 treated lesions in a total of 7 horses. Complete disappearance was observed in one treated lesion, with no recurrence after 6 months. No adverse events have been observed in any of the animals during and after treatment. These results demonstrate the effectiveness and safety of interleukin 12 encoding plasmid DNA therapy against established metastatic disease in a large animal model and serve as a basis for a clinical trial. PMID- 11434723 TI - Antibody recognition of amyloidogenic transthyretin variants in serum of patients with familial amyloidotic polyneuropathy. AB - Familial amyloidotic polyneuropathy (FAP) is a late-onset inherited disease characterized by the deposition of amyloid fibrils. FAP is associated with mutations on the transthyretin (TTR) gene. A monoclonal antibody, MAb 39-44, reacting with high molecular weight aggregates of TTR but not with tetrameric TTR has recently been generated and characterized. This antibody recognizes a cryptic epitope that is expressed in isolated recombinant amyloidogenic mutants and in ex vivo amyloid. In the present work we show that this amyloid-specific antibody specifically recognizes in a direct enzyme-linked immunoassay (ELISA) plasma TTR from carriers of various mutations associated with FAP, both in asymptomatic individuals and in patients. In contrast, it does not react with plasma TTR from healthy individuals or that from carriers of nonpathogenic mutations. Using the ELISA developed in this study we identified three different TTR mutations in Portuguese patients with neuropathy of unknown cause, later shown to have amyloid tissue deposition. This antibody recognizes conformations that express cryptic epitopes shared by amyloidogenic TTR variants associated with FAP, not present among nonpathogenic TTR molecules. This antibody will contribute to further identify and characterize intermediates of the amyloidogenic cascade. In addition, it will also be useful for screening amyloidogenic TTR mutations in patients with neuropathy of unknown cause, prior to precise molecular diagnosis using protein and/or DNA analysis. PMID- 11434724 TI - Natural zeolite clinoptilolite: new adjuvant in anticancer therapy. AB - Natural silicate materials, including zeolite clinoptilolite, have been shown to exhibit diverse biological activities and have been used successfully as a vaccine adjuvant and for the treatment of diarrhea. We report a novel use of finely ground clinoptilolite as a potential adjuvant in anticancer therapy. Clinoptilolite treatment of mice and dogs suffering from a variety of tumor types led to improvement in the overall health status, prolongation of life-span, and decrease in tumors size. Local application of clinoptilolite to skin cancers of dogs effectively reduced tumor formation and growth. In addition, toxicology studies on mice and rats demonstrated that the treatment does not have negative effects. In vitro tissue culture studies showed that finely ground clinoptilolite inhibits protein kinase B (c-Akt), induces expression of p21WAF1/CIP1 and p27KIP1 tumor suppressor proteins, and blocks cell growth in several cancer cell lines. These data indicate that clinoptilolite treatment might affect cancer growth by attenuating survival signals and inducing tumor suppressor genes in treated cells. PMID- 11434725 TI - Large de novo deletion in chromosome 12 affecting the PAH, IGF1, ASCL1, and TRA1 genes. AB - Phenylketonuria is one of the most common genetic diseases in humans, affecting 1 in 10,000 whites. Deletions are generally uncommon in genes in which no long highly homologous segments are present, and in phenylalanine hydroxylase (PAH) deficiency they represent only 5% of cases. We present the case of a girl affected by classical phenylketonuria who has been screened for mutations in the PAH gene. During the molecular study a large de novo deletion has detected in 12qter, including PAH, and the genes for insulin-like growth factor 1 (IGF1), human achaete-scute homolog 1 (ASCL1), and tumor rejection antigen (TRA1). The patient showed phenylketonuria, short stature, and pathological electro oculography results in both eyes, with high affectation of the relative electrogenesis of the photoreceptor-pigment epithelium complex. She had previously been misdiagnosed as homozygous for the IVS8nt-7A-G mutation, instead of heterozygous for a mutation and a de novo deletion. As a result incorrect genetic counseling had been given. The deletion of the PAH, IGF1, and ASCL1 genes could explain the patient's phenotype corresponding to a contiguous gene syndrome. We stress the relevance of polymorphic marker haplotype analysis and the importance of family study in genetic recessive diseases, such as phenylketonuria, to avoid incorrect diagnosis and genetic counseling. PMID- 11434726 TI - Postmenopausal hormone use and secondary prevention of coronary events in the nurses' health study. a prospective, observational study. AB - BACKGROUND: The Heart and Estrogen/progestin Replacement Study (HERS) was the first randomized clinical trial of combined hormone therapy and secondary prevention of coronary events. The trial had overall null results but reported an unexpected increased risk for recurrent events in the initial year, followed by a decrease during the final years. OBJECTIVE: To provide additional data on a time trend in risk for recurrent heart disease. DESIGN: A prospective, observational cohort study of secondary prevention of coronary heart disease. SETTING: Nurses' Health Study. PATIENTS: 2489 postmenopausal women with previous myocardial infarction or documented atherosclerosis; 213 cases of recurrent nonfatal myocardial infarction or coronary death were identified from 1976 through 1996. MEASUREMENTS: Information on hormone status and on recurrent disease was collected by using biennial questionnaires. Multivariable-adjusted relative risks and 95% CIs were calculated from logistic regression models. RESULTS: A trend of decreasing risk for recurrent major coronary heart disease events with increasing duration of hormone use was observed (P for trend = 0.002). For short-term current users, the multivariate-adjusted relative risk for major coronary heart disease was 1.25 (95% CI, 0.78 to 2.00) compared with never-users. However, after longer-term hormone use, the rate of second events was lower in current users than in never-users (relative risk, 0.38 [CI, 0.22 to 0.66]). No clear differences emerged between users of estrogen alone and users of estrogen combined with progestin. Overall, with up to 20 years of follow-up, the relative risk for a second event among current users of hormone therapy was 0.65 (CI, 0.45 to 0.95) compared with never-users. CONCLUSIONS: The risk for recurrent major coronary events seems to increase among short-term hormone users with previous coronary disease but to decrease with longer-term use. PMID- 11434727 TI - Patient-initiated treatment of uncomplicated recurrent urinary tract infections in young women. AB - BACKGROUND: Recurrent urinary tract infections (UTIs) are a common outpatient problem, resulting in frequent office visits and often requiring the use of prophylactic antimicrobial agents. Patient-initiated treatment of recurrent UTIs may decrease antimicrobial use and improve patient convenience. OBJECTIVE: To determine the safety and feasibility of patient-initiated treatment of recurrent UTIs. DESIGN: Uncontrolled, prospective clinical trial. SETTING: University-based primary health care clinic. PARTICIPANTS: Women at least 18 years of age with a history of recurrent UTIs and no recent pregnancy, hypertension, diabetes, or renal disease. INTERVENTION: After self-diagnosing UTI on the basis of symptoms, participating women initiated therapy with ofloxacin or levofloxacin. MEASUREMENTS: Accuracy of self-diagnosis determined by evidence of a definite (culture-positive) or probable (sterile pyuria and no alternative diagnosis) UTI on pretherapy urinalysis and culture. Women with a self-diagnosis of UTI that was not microbiologically confirmed were evaluated for alternative diagnoses. Post therapy interviews and urine cultures were used to assess clinical and microbiological cure rates, adverse events, and patient satisfaction. RESULTS: 88 of 172 women self-diagnosed a total of 172 UTIs. Laboratory evaluation showed a uropathogen in 144 cases (84%), sterile pyuria in 19 cases (11%), and no pyuria or bacteriuria in 9 cases (5%). Clinical and microbiological cures occurred in 92% and 96%, respectively, of culture-confirmed episodes. No serious adverse events occurred. CONCLUSION: Adherent women can accurately self-diagnose and self treat recurrent UTIs. PMID- 11434728 TI - Highly active antiretroviral therapy decreases mortality and morbidity in patients with advanced HIV disease. AB - BACKGROUND: Mortality and morbidity related to AIDS have decreased among HIV infected patients taking highly active anti-retroviral therapy (HAART), but previous studies may have been confounded by other changes in treatment. OBJECTIVE: To assess the benefit of HAART in patients with advanced AIDS and anemia. DESIGN: Prospective, multicenter cohort study. SETTING: The Viral Activation Transfusion Study (VATS), with enrollment from August 1995 through July 1998 and follow-up through June 1999. PATIENTS: 528 HIV-infected patients with cytomegalovirus (CMV) seropositivity or disease who were receiving a first red blood cell transfusion for anemia. MEASUREMENTS: In a person-year analysis of follow-up before and after initiation of HAART, Poisson regression was used to calculate crude rate ratios and rate ratios adjusted for CD4 count, HIV RNA level, calendar period, time on study, sex, ethnicity, and injection drug use. RESULTS: At baseline, patients had a median CD4(+) lymphocyte count of 0.015 x 10(9) cell/L, median plasma HIV RNA level of 4.8 log(10) copies/mL, and median hemoglobin concentration of 73 g/L. Use of HAART increased from 1% of active patients in January 1996 to 79% of active patients in January 1999. The crude death rate was 0.24 event/person-year among patients taking HAART and 0.88 event/person-year among those not taking HAART (rate ratio, 0.26; adjusted rate ratio, 0.38; P < 0.001 for both comparisons). Rates of non-CMV disease were 0.15 event/ person-year after HAART and 0.45 event/person-year before HAART (crude rate ratio, 0.34 [ P < 0.001]; adjusted rate ratio, 0.66 [ P < 0.05]). Rates of CMV disease were 0.10 event/person-year after HAART and 0.25 before HAART (crude rate ratio, 0.42 [ P < 0.01]; adjusted rate ratio, 1.01 [ P > 0.2]). Results were similar in patients with baseline CD4(+) lymphocyte counts less than 0.010 x 10(9) cells/L. CONCLUSIONS: The data support an independent reduction in mortality and opportunistic events attributable to HAART, even in patients with very advanced HIV disease. However, patients with CMV infection or disease may not have a reduction in new CMV events due to HAART. PMID- 11434729 TI - Treatment of complicated sarcoidosis with infliximab anti-tumor necrosis factor alpha therapy. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) may have an important role in the clinical exacerbation of sarcoidosis. OBJECTIVE: To treat sarcoidosis with infliximab, a chimeric human-murine anti-human TNF-alpha monoclonal antibody. DESIGN: Case report. SETTING: U.S. academic medical center. PATIENT: A 72-year old woman with sarcoidosis presenting with severe protein-losing enteropathy, hypoalbuminemia, and proximal myopathy who had not responded adequately to corticosteroid therapy and whose clinical course was further complicated by acute tubular necrosis and renal failure requiring long-term hemodialysis. INTERVENTION: Intravenous infusion of infliximab, 5 mg/kg of ideal body weight; infusion was repeated at 2 and 6 weeks. MEASUREMENTS: Clinical response of enteropathic and myopathic symptoms and serum albumin level. RESULTS: Enteropathic and myopathic symptoms resolved after infliximab therapy, and the serum albumin level also improved. However, the clinical course was complicated by the development of a hypercoagulable state associated with circulating anticardiolipin antibodies, which prompted discontinuation of infliximab therapy. CONCLUSIONS: Infliximab therapy was successful in a patient with sarcoidosis. Tumor necrosis factor-alpha may be an important mediator of clinical disease in sarcoidosis and could be an attractive target for therapeutic intervention. However, infliximab may cause adverse effects associated with cytokine cascade manipulation. PMID- 11434730 TI - Diagnosis and management of delirium near the end of life. AB - Delirium is a common and distressing symptom that constitutes a significant challenge for end-of-life care. However, reliable techniques are available for the diagnosis of delirium, and effective therapies exist as well. This consensus paper uses a case-based format that begins with an overview of the definition and presentation of delirium. Next, strategies for diagnosis are suggested, with attention to the unique challenges that clinicians face in pursuing a diagnostic work-up for patients near the end of life. The paper concludes with a review of therapeutic options. PMID- 11434731 TI - Increasing antimicrobial resistance and the management of uncomplicated community acquired urinary tract infections. AB - Community-acquired urinary tract infections (UTIs) are among the most common bacterial infections in women. Therapy for these infections is usually begun before results of microbiological tests are known. Furthermore, in women with acute uncomplicated cystitis, empirical therapy without a pretherapy urine culture is often used. The rationale for this approach is based on the highly predictable spectrum of etiologic agents causing UTI and their antimicrobial resistance patterns. However, antimicrobial resistance among uropathogens causing community-acquired UTIs, both cystitis and pyelonephritis, is increasing. Most important has been the increasing resistance to trimethoprim-sulfamethoxazole (TMP-SMX), the current drug of choice for treatment of acute uncomplicated cystitis in women. What implications do these trends have for treatment of community-acquired UTIs? Preliminary data suggest that clinical cure rates may be lower among women with uncomplicated cystitis treated with TMP-SMX when the infecting pathogen is resistant to TMP-SMX. Women with pyelonephritis also have less bacterial eradication and lower clinical cure rates when treated with TMP SMX for an infection that is resistant to the drug. Therefore, in the outpatient setting, identifying risk factors for TMP-SMX resistance and knowing the prevalence of TMP-SMX resistance in the local community are important steps in choosing an appropriate therapeutic agent. When choosing a treatment regimen, physicians should consider such factors as in vitro susceptibility, adverse effects, cost-effectiveness, and selection of resistant strains. Using a management strategy that takes these variables into account is essential for maintaining the safety and efficacy of treatment for acute UTI. PMID- 11434732 TI - Helping patients take responsibility for their own health. PMID- 11434733 TI - Looking forward at Annals. PMID- 11434734 TI - What the stethoscope said. PMID- 11434735 TI - A presentation at the weekly meeting of the Institutional Review Board. PMID- 11434738 TI - In defense of a department of geriatrics. PMID- 11434740 TI - Placebo-controlled trials. PMID- 11434741 TI - Placebo-controlled trials. PMID- 11434743 TI - Update in infectious diseases. PMID- 11434745 TI - Testosterone and resistance training in AIDS. PMID- 11434747 TI - Garlic for total cholesterol reduction. PMID- 11434749 TI - Wine and mortality. PMID- 11434751 TI - Smoking and abnormalities in renal function. PMID- 11434752 TI - Smoking and abnormalities in renal function. PMID- 11434754 TI - Kava hepatotoxicity. PMID- 11434755 TI - Medication assistance programs. PMID- 11434756 TI - Acute renal failure related to high-dose celecoxib. PMID- 11434757 TI - Usefulness of online Mendelian Inheritance in Man in clinical practice. PMID- 11434758 TI - Idiopathic thrombocytopenic purpura in a 101-year-old woman. PMID- 11434759 TI - Correction: physicians and joint negotiation. PMID- 11434763 TI - Current topics in RNA-protein recognition: control of specificity and biological function through induced fit and conformational capture. PMID- 11434764 TI - Increased backbone flexibility in threonine45-phosphorylated hirudin upon pH change. AB - Protein phosphorylation on serine/threonine side chains represents a major regulatory event in the posttranslational control of protein functionality, where it is thought to operate at the level of structural changes in the polypeptide chain. However, key questions about molecular aspects of phosphate ester induced conformational alterations remain open. Among these concerns are the radius of action of the phosphate ester group, its effective ionic state, and its interplay with distinct bonds of the polypeptide chain. Primarily to define short-range effects upon threonine phosphorylation, the native 65 amino acid protein hirudin, conformationally restrained by a proline flanking the pThr(45) site and three intramolecular disulfide bonds, was structurally characterized in both the phosphorylated and the unphosphorylated state in solution. Circular dichroism and hydrogen exchange experiments (MALDI-TOF) showed that structural changes were caused by Thr(45)-Pro(46) phosphorylation only when the phosphate ester group was in its dianionic state. The spatial arrangement of the amino acids, monitored by 1H NMR spectroscopy, appears to be affected within a radius of about 10 A around the pThr(45)-OgammaH, with phosphorylation resulting in a loss of structure and increased flexibility within a segment of at least seven amino acid residues. Thus, the transition from the monoanionic to the dianionic phosphate group over the pH range 5.2-8.5 represents a general phosphorylation-dependent conformational switch operating at physiological pH values. PMID- 11434765 TI - Biosynthesis of D-alanyl-lipoteichoic acid: the tertiary structure of apo-D alanyl carrier protein. AB - The D-alanylation of lipoteichoic acid (LTA) allows the Gram-positive organism to modulate its surface charge, regulate ligand binding, and control the electromechanical properties of the cell wall. The incorporation of D-alanine into LTA requires the D-alanine:D-alanyl carrier protein ligase (AMP-forming) (Dcl) and the carrier protein (Dcp). The high-resolution solution structure of the 81-residue (8.9 kDa) Dcp has been determined by multidimensional heteronuclear NMR. An ensemble of 30 structures was calculated using the torsion angle dynamics approach of DYANA. These calculations utilized 3288 NOEs containing 1582 unique nontrivial NOE distance constraints. Superposition of residues 4-81 on the mean structure yields average atomic rmsd values of 0.43 +/- 0.08 and 0.86 +/- 0.09 A for backbone and non-hydrogen atoms, respectively. The solution structure is composed of three alpha-helices in a bundle with additional short 3(10)- and alpha-helices in intervening loops. Comparisons of the three dimensional structure with the acyl carrier proteins involved in fatty acid, polyketide, and nonribosomal peptide syntheses support the conclusion that Dcp is a homologue in this family. While there is conservation of the three-helix bundle fold, Dcp has a higher enthalpy of unfolding and no apparent divalent metal binding site(s), features that distinguish it from the fatty acid synthase acyl carrier protein of Escherichia coli. This three-dimensional structure also provides insights into the D-alanine ligation site recognized by Dcl, as well as the site which may bind the poly(glycerophosphate) acceptor moiety of membrane associated LTA. PMID- 11434766 TI - Solution structure of the squash trypsin inhibitor MCoTI-II. A new family for cyclic knottins. AB - The "knottin" fold is a stable cysteine-rich scaffold, in which one disulfide crosses the macrocycle made by two other disulfides and the connecting backbone segments. This scaffold is found in several protein families with no evolutionary relationships. In the past few years, several homologous peptides from the Rubiaceae and Violaceae families were shown to define a new structural family based on macrocyclic knottin fold. We recently isolated from Momordica cochinchinensis seeds the first known macrocyclic squash trypsin inhibitors. These compounds are the first members of a new family of cyclic knottins. In this paper, we present NMR structural studies of one of them, MCoTI-II, and of a beta Asp rearranged form, MCoTI-IIb. Both compounds display similar and well-defined conformations. These cyclic squash inhibitors share a similar conformation with noncyclic squash inhibitors such as CPTI-II, and it is postulated that the main effect of the cyclization is a reduced sensitivity to exo-proteases. On the contrary, clear differences were detected with the three-dimensional structures of other known cyclic knottins, i.e., kalata B1 or circulin A. The two-disulfide cystine-stabilized beta-sheet motif [Heitz et al. (1999) Biochemistry 38, 10615 10625] is conserved in the two families, whereas in the C-to-N linker, one disulfide bridge and one loop are differently located. The molecular surface of MCoTI-II is almost entirely charged in contrast to circulin A that displays a well-marked amphiphilic character. These differences might explain why the isolated macrocyclic squash inhibitors from M. cochinchinensis display no significant antibacterial activity, whereas circulins and kalata B1 do. PMID- 11434767 TI - Mossbauer and EPR studies of the photoactivation of nitrile hydratase. AB - The alphabeta dimer of active nitrile hydratase from Rhodococcus sp. R312 contains one low-spin ferric ion that is coordinated by three Cys residues, two N amide groups from the protein backbone, and one OH(-). The enzyme isolated from bacteria grown in the dark is inactive and contains the iron site as a six coordinate diamagnetic Fe-nitrosyl complex, called NH(dark). The active state can be obtained from the dark state by photolysis of the Fe-NO bond at room temperature. Activation is accompanied by the conversion of NH(dark) to a low spin ferric complex, NH(light), exhibiting an S = (1)/(2) EPR signal with g values of 2.27, 2.13, and 1.97. We have characterized both NH(dark) and NH(light) with Mossbauer spectroscopy. The z-axis of the 57Fe magnetic hyperfine tensor, A, of NH(light) was found to be rotated by approximately 45 degrees relative to the z-axis of the g tensor (g(z) = 1.97). Comparison of the A tensor of NH(light) with the A tensors of low-spin ferric hemes indicates a substantially larger degree of covalency for nitrile hydratase. We have also performed photolysis experiments between 2 and 20 K and characterized the photolyzed products by EPR and Mossbauer spectroscopy. Photolysis at 4.2 K in the Mossbauer spectrometer yielded a five-coordinate low-spin ferric species, NH(A), which converted back into NH(dark) when the sample was briefly warmed to 77 K. We also describe preliminary EPR photolysis studies that have yielded new intermediates. PMID- 11434768 TI - Inhibition of AmpC beta-lactamase through a destabilizing interaction in the active site. AB - Beta-lactamases hydrolyze beta-lactam antibiotics, including penicillins and cephalosporins; these enzymes are the most widespread resistance mechanism to these drugs and pose a growing threat to public health. beta-Lactams that contain a bulky 6(7)alpha substituent, such as imipenem and moxalactam, actually inhibit serine beta-lactamases and are widely used for this reason. Although mutant serine beta-lactamases have arisen that hydrolyze beta-lactamase resistant beta lactams (e.g., ceftazidime) or avoid mechanism-based inhibitors (e.g., clavulanate), mutant serine beta-lactamases have not yet arisen in the clinic with imipenemase or moxalactamase activity. Structural and thermodynamic studies suggest that the 6(7)alpha substituents of these inhibitors form destabilizing contacts within the covalent adduct with the conserved Asn152 in class C beta lactamases (Asn132 in class A beta-lactamases). This unfavorable interaction may be crucial to inhibition. To test this destabilization hypothesis, we replaced Asn152 with Ala in the class C beta-lactamase AmpC from Escherichia coli and examined the mutant enzyme's thermodynamic stability in complex with imipenem and moxalactam. Consistent with the hypothesis, the Asn152 --> Ala substitution relieved 0.44 and 1.10 kcal/mol of strain introduced by imipenem and moxalactam, respectively, relative to the wild-type complexes. However, the kinetic efficiency of AmpC N152A was reduced by 6300-fold relative to that of the wild type enzyme. To further investigate the inhibitor's interaction with the mutant enzyme, the X-ray crystal structure of moxalactam in complex with N152A was determined to a resolution of 1.83 A. Moxalactam in the mutant complex is significantly displaced from its orientation in the wild-type complex; however, moxalactam does not adopt an orientation that would restore competence for hydrolysis. Although Asn152 forces beta-lactams with 6(7)alpha substituents out of a catalytically competent configuration, making them inhibitors, the residue is essential for orienting beta-lactam substrates and cannot simply be replaced with a much smaller residue to restore catalytic activity. Designing beta-lactam inhibitors that interact unfavorably with this conserved residue when in the covalent adduct merits further investigation. PMID- 11434769 TI - Microtubule structure at improved resolution. AB - Microtubule architecture can vary with eukaryotic species, with different cell types, and with the presence of stabilizing agents. For in vitro assembled microtubules, the average number of protofilaments is reduced by the presence of sarcodictyin A, epothilone B, and eleutherobin (similarly to taxol) but increased by taxotere. Assembly with a slowly hydrolyzable GTP analogue GMPCPP is known to give 96% 14 protofilament microtubules. We have used electron cryomicroscopy and helical reconstruction techniques to obtain three-dimensional maps of taxotere and GMPCPP microtubules incorporating data to 14 A resolution. The dimer packing within the microtubule wall is examined by docking the tubulin crystal structure into these improved microtubule maps. The docked tubulin and simulated images calculated from "atomic resolution" microtubule models show tubulin heterodimers are aligned head to tail along the protofilaments with the beta subunit capping the microtubule plus end. The relative positions of tubulin dimers in neighboring protofilaments are the same for both types of microtubule, confirming that conserved lateral interactions between tubulin subunits are responsible for the surface lattice accommodation observed for different microtubule architectures. Microtubules with unconventional protofilament numbers that exist in vivo are likely to have the same surface lattice organizations found in vitro. A curved "GDP" tubulin conformation induced by stathmin-like proteins appears to weaken lateral contacts between tubulin subunits and could block microtubule assembly or favor disassembly. We conclude that lateral contacts between tubulin subunits in neighboring protofilaments have a decisive role for microtubule stability, rigidity, and architecture. PMID- 11434770 TI - Role of metal ions in catalysis by enolase: an ordered kinetic mechanism for a single substrate enzyme. AB - Spectroscopic and kinetic methods have been used to explore the roles of divalent metal ions in the enolase-catalyzed dehydration of 2-phosphoglycerate (2-PGA). Enolase requires 2 equiv of metal ion per active site for maximal activity. Previous crystallographic studies [Larsen, T. M., Wedekind, J. E., Rayment, I., and Reed, G. H. (1996) Biochemistry 35, 4349-4358] showed that both magnesium ions coordinated to the carboxylate group of the substrate/product-a scheme consistent with metal ion assistance in formation of the enolate intermediate. Electron paramagnetic resonance (EPR) data with 17O-labeled forms of phosphoenolpyruvate show that Mn(2+), bound at the lower affinity site, coordinates to one carboxylate oxygen and one phosphate oxygen of the substrate. These observations are fully consistent with the crystallographic data. Plots of activity versus log [metal ion] are bell-shaped, and the inhibitory phases of the profiles have been previously attributed to binding of metal ions at ancillary sites on the enzyme. However, the activation profiles and measurements of 2H kinetic isotope effects support an ordered kinetic mechanism wherein binding of 2 PGA precedes binding of the second metal ion, and release of the second metal ion occurs prior to departure of phosphoenolpyruvate. High concentrations of metal ion lead to inhibition in the ordered mechanism by interfering with product release. The 2H kinetic isotope effect is diminished in the inhibitory phases of the metal ion activation profiles in a manner that is consistent with the predominantly ordered mechanism. Zn(2+) gives lower maximal activity than Mg(2+), apparently due to slow release of Zn(2+) from the product complex. Addition of imidazole increases the maximal rate apparently by accelerating the release of Zn(2+) from the enzyme. PMID- 11434771 TI - Comparison of the amino acid residues in the sixth transmembrane domains accessible in the binding-site crevices of mu, delta, and kappa opioid receptors. AB - We have mapped the residues in the sixth transmembrane domains (TMs 6) of the mu, delta, and kappa opioid receptors that are accessible in the binding-site crevices by the substituted cysteine accessibility method (SCAM). We previously showed that ligand binding to the C7.38S mutants of the mu and kappa receptors and the wild-type delta receptor was relatively insensitive to methanethiosulfonate ethylammonium (MTSEA), a positively charged sulfhydryl specific reagent. These MTSEA-insensitive constructs were used as the templates, and 22 consecutive residues in TM6 (excluding C6.47) of each receptor were mutated to cysteine, 1 at a time. Most mutants retained binding affinities for [3H]diprenorphine, a nonselective opioid antagonist, similar to that of the template receptors. Treatment with MTSEA significantly inhibited [3H]diprenorphine binding to 11 of 22 mutants of the rat mu receptor and 9 of 22 mutants of the human delta receptor and 10 of 22 mutants of the human kappa receptor. Naloxone or diprenorphine protected all sensitive mutants, except the A6.42(287)C mu mutant. Thus, V6.40, F6.44, W6.48, I6.51, Y6.54, V6.55, I6.56, I6.57, K6.58, and A6.59 of the mu receptor; F6.44, I6.51, F6.54, V6.55, I6.56, V6.57, W6.58, T6.59, and L6.60 of the delta receptor; and F6.44, W6.48, I6.51, F6.54, I6.55, L6.56, V6.57, E6.58, A6.59, and L6.60 of the kappa receptor are on the water-accessible surface of the binding-site crevices. The accessibility patterns of residues in the TMs 6 of the mu, delta, and kappa opioid receptors are consistent with the notion that the TMs 6 are in alpha-helical conformations with a narrow strip of accessibility on the intracellular side of 6.54 and a wider area of accessibility on the extracellular side of 6.54, likely due to a proline kink at 6.50 that bends the helix in toward the binding pocket and enables considerable motion in this region. The wider exposure of residues 6.55 6.60 to the binding-site crevice, combined with the divergent amino acid sequences, is consistent with the inferred role of residues in this region in determining ligand binding selectivity. The conservation of the accessibility pattern on the cytoplasmic side of 6.54 suggests that this region may be important for receptor activation. This accessibility pattern is similar to that of the D2 dopamine receptor, the only other GPCR in which TM6 has been mapped by SCAM. That opioid receptors and the remotely related D2 dopamine receptor have similar accessibility patterns in TM6 suggest that these segments of GPCRs in the rhodopsin-like subfamily not only share secondary structure but also are packed similarly into the transmembrane bundle and thus have similar tertiary structure. PMID- 11434772 TI - Structural role of the proline residues of the beta-hinge region of p13suc1 as revealed by site-directed mutagenesis and fluorescence studies. AB - Site-directed mutagenesis, gel filtration, and fluorescence spectroscopy approaches were used to study the molecular hinge mechanism involved in the beta strand-exchanged dimer formation of the cyclin-dependent protein kinase regulatory subunit p13(suc1) from Schizosaccharomyces pombe. Single and double mutants of residues Pro-90 and Pro-92 (P90V, P92V, and P90V/P92V) were prepared and assayed. Substitution of Pro-90 prevented dimer formation by arm exchange. However, single point mutations did not affect the two-state unfolding transition of wild-type p13(suc1) at equilibrium (i.e., wild type, DeltaG degrees (0,un) = 7.38 +/- 0.35 kcal mol(-1), vs P90V, DeltaG degrees (0,un) = 6.71 +/- 0.18 kcal mol(-1)). On the contrary, the double mutant unfolded with a complex transition, and the reaction was best described by a three-state model (N <==> I <==> U). Resolution of the state-dependent (native vs denatured) intrinsic fluorescence decay amplitudes of p13(suc1) showed that with P90V/P92V these parameters were affected at [GuHCl] significantly less than with wild-type and single mutant proteins. Moreover, with the latter products, fluorescence quenching measurements at 1 M GuHCl revealed linear Stern-Volmer plots with quenching constants typical of tryptophan residues located in a native environment (1.6 M(-1) < K(SV) < 2.3 M(-1)). Dissimilarly, with P90V/P92V a significant deviation from linearity of the Stern-Volmer plot was obtained. Nonlinear least-squares analysis of these data resolved the significant contribution of highly solvent-accessible emitting species (K(SV) = 26 M(-1)) consistent with large exposure of the tryptophan residues. These results are compatible with the existence of an intermediate unfolding state of the double mutation product. Thus, while single residue substitution studies give support to the primary role of Pro-90 in the p13(suc1) dimer formation by domain swapping, double residue substitution studies indicate the important role of the conserved repeat, Pro-x-Pro, for the proper beta-strand spatial organization and stability. PMID- 11434773 TI - A transition-state analogue reduces protein dynamics in hypoxanthine-guanine phosphoribosyltransferase. AB - Hypoxanthine-guanine phosphoribosyltransferase (HGPRT) is the key enzyme in purine base salvage in humans and in purine auxotrophs, including Plasmodium falciparum, the leading cause of malaria. Hydrogen/deuterium (H/D) exchange into amide bonds, quantitated by on-line HPLC and mass spectrometry, has been used to compare the dynamic and conformational properties of human HGPRT alone, the HGPRT GMP-Mg(2+) complex, the HGPRT-IMP-MgPPi <==> HGPRT-Hx-MgPRPP equilibrating mixture, and the transition-state analogue complex HGPRT-ImmGP-MgPPi. The rate and extent of H/D exchange of 26 peptic peptides, spanning 91% of the primary structure, have been monitored. Human HGPRT has 207 amide H/D exchange sites. After 1 h in D2O, HGPRT alone exchanges 160, HGPRT-GMP-Mg(2+) exchanges 154, the equilibrium complex exchanges 139, and the transition-state analogue complex exchanges 126 of these amide protons. H/D exchange rates are correlated with structure for peptides in (1) catalytic site loops, (2) a connected peptide of the subunit interface of the tetramer, and (3) a loop buried in the catalytic site. Structural properties related to H/D exchange are defined from crystallographic studies of the HGPRT-GMP-Mg(2+) and HGPRT-ImmGP-MgPPi complexes. Transition-state analogue binding strengthens the interaction between subunits and tightens the catalytic site loops. The solvent exchange dynamics in specific peptides correlates with hydrogen bond patterns, solvent access, crystallographic B-factors, and ligand exchange rates. Solvent exchange reveals loop dynamics in the free enzyme, Michaelis complexes, and the complex with the bound transition state analogue. Proton transfer paths, rather than dynamic motion, are required to explain exchange into a buried catalytic site peptide in the complex with the bound transition-state analogue. PMID- 11434774 TI - Structure of an RNA hairpin from HRV-14. AB - The 5' noncoding region of the picornaviral genome begins with a cloverleaf which is required for viral replication, due at least in part to an interaction with the viral RNA polymerase as part of a fusion with the predominant viral protease. The necessary region of the cloverleaf has previously been narrowed to a highly conserved stem-loop. The solution structure of a 14-nucleotide RNA hairpin, which is part of the conserved stem-loop from human rhinovirus isotype 14, is presented here. The secondary structure of the hairpin is identical to predictions: a five base pair stem is bounded by a triloop with sequence UAU. However, the fold of the triloop is novel, with stacking of the second loop base onto the closing base pair of the stem, and deviations from A form geometry are introduced into the stem regions bordering the triloop, particularly on the 3' side. These deviations and the associated triloop structure could help to explain the distinct sequence conservation and mutational analysis data observed for the stem region of the hairpin, as compared to a second sequentially similar stem in the intact stem loop. PMID- 11434775 TI - Phospholipid scramblase activation pathways in lymphocytes. AB - In erythrocytes and platelets, activation of a nonspecific lipid flipsite termed the scramblase allows rapid, bidirectional transbilayer movement of all types of phospholipids. When applied to lymphoid cells, scramblase assays reveal a similar activity, with scrambling rates intermediate between those seen in platelets and erythrocytes. Scrambling activity initiated in lymphoid cells by elevation of intracellular Ca(2+) proceeds after a lag not noted in platelets or erythrocytes. The rates of transbilayer movement of phosphatidylserine and phosphatidylcholine analogues are similar whether the scramblase is activated by elevated internal Ca(2+) or by apoptosis. Elevation of internal Ca(2+) levels in apoptotic cells does not result in an additive increase in the rate of lipid movement. In lymphoid cells from a patient with Scott syndrome, scramblase cannot be activated by Ca(2+), but is induced normally during apoptosis. These findings suggest that Ca(2+) and apoptosis operate through different pathways to activate the same scramblase. PMID- 11434776 TI - Copper and zinc binding modulates the aggregation and neurotoxic properties of the prion peptide PrP106-126. AB - The abnormal form of the prion protein (PrP) is believed to be responsible for the transmissible spongiform encephalopathies. A peptide encompassing residues 106-126 of human PrP (PrP106-126) is neurotoxic in vitro due its adoption of an amyloidogenic fibril structure. The Alzheimer's disease amyloid beta peptide (Abeta) also undergoes fibrillogenesis to become neurotoxic. Abeta aggregation and toxicity is highly sensitive to copper, zinc, or iron ions. We show that PrP106-126 aggregation, as assessed by turbidometry, is abolished in Chelex-100 treated buffer. ICP-MS analysis showed that the Chelex-100 treatment had reduced Cu(2+) and Zn(2+) levels approximately 3-fold. Restoring Cu(2+) and Zn(2+) to their original levels restored aggregation. Circular dichroism showed that the Chelex-100 treatment reduced the aggregated beta-sheet content of the peptide. Electron paramagnetic resonance spectroscopy identified a 2N1S1O coordination to the Cu(2+) atom, suggesting histidine 111 and methionine 109 or 112 are involved. Nuclear magnetic resonance confirmed Cu(2+) and Zn(2+) binding to His-111 and weaker binding to Met-112. An N-terminally acetylated PrP106-126 peptide did not bind Cu(2+), implicating the free amino group in metal binding. Mutagenesis of either His-111, Met-109, or Met-112 abolished PrP106-126 neurotoxicity and its ability to form fibrils. Therefore, Cu(2+) and/or Zn(2+) binding is critical for PrP106-126 aggregation and neurotoxicity. PMID- 11434777 TI - Bruton's tyrosine kinase is essential for hydrogen peroxide-induced calcium signaling. AB - Using Btk-deficient DT40 cells and the transfectants expressing wild-type Btk or Btk mutants in either kinase (Arg(525) to Gln), Src homology 2 (SH2, Arg(307) to Ala), or pleckstrin homology (PH, Arg(28) to Cys) domains, we investigated the roles and structure-function relationships of Btk in hydrogen peroxide-induced calcium mobilization. Our genetic evidence showed that Btk deficiency resulted in a significant reduction in hydrogen peroxide-induced calcium response. This impaired calcium signaling is correlated with the complete elimination of IP3 production and the significantly reduced tyrosine phosphorylation of PLCgamma2 in Btk-deficient DT40 cells. All of these defects were fully restored by the expression of wild-type Btk in Btk-deficient DT40 cells. The data from the point mutation study revealed that a defect at any one of the three functional domains would prevent a full recovery of Btk-mediated hydrogen peroxide-induced intracellular calcium mobilization. However, mutation at either the SH2 or PH domain did not affect the hydrogen peroxide-induced activation of Btk. Mutation at the SH2 domain abrogates both IP3 generation and calcium release, while the mutant with the nonfunctional PH domain can partially activate PLCgamma2 and catalyze IP3 production but fails to produce significant calcium mobilization. Thus, these observations suggest that Btk-dependent tyrosine phosphorylation of PLCgamma2 is required but not sufficient for hydrogen peroxide-induced calcium mobilization. Furthermore, hydrogen peroxide stimulates a Syk-, but not Btk-, dependent tyrosine phosphorylation of B cell linker protein BLNK. The overall results, together with those reported earlier [Qin et al. (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 7118], are consistent with the notion that functional SH2 and PH domains are required for Btk to form a complex with PLCgamma2 through BLNK in order to position the Btk, PLCgamma2, and phosphatidylinositol 4,5 bisphosphate in close proximity for efficient activation of PLCgamma2 and to maximize its catalytic efficiency for IP3 production. PMID- 11434778 TI - Flexible loop of beta 2-glycoprotein I domain V specifically interacts with hydrophobic ligands. AB - Beta2-glycoprotein I (beta2-GPI), which consists of four complement control protein modules and a distinctly folded fifth C-terminal domain, is an essential cofactor for the binding to phospholipids of anti-cardiolipin antibodies, isolated from patients with anti-phospholipid antibody syndrome, and its fifth domain has attracted attention as a specific phospholipid-binding site. We focused on the fifth domain of beta2-GPI (Domain V) and examined the interaction of intact Domain V, Domains IV-V, and nicked Domain V with various hydrophobic ligands, as a model molecule of phospholipid. We found that electrostatic and hydrophobic interactions are important for Domain V binding to the ligand molecules. We also found that, while Domain IV has no significant effect on the interactions with ligands, the nicked Domain V with cleavage in the flexible loop decreases the affinity, indicating that the flexible loop region is the binding site of the hydrophobic ligands. The synthetic peptide corresponding to the loop region was disordered and interacted with bis-ANS, confirming the critical role of the loop region. To clarify the nature of the interaction between the loop region and hydrophobic compounds, we prepared the reduced and alkylated Domain V, which was denatured but was assumed to be a collapsed state. Alkylation by iodoacetic acid decreased the interaction of Domain V with bis-ANS, probably because the protein net charge was decreased by the six introduced carboxyl groups and consequently the electrostatic interactions were decreased. In contrast, Domain V alkylated by iodoacetamide, therefore retaining a high positive net charge, bound bis-ANS more strongly than the intact Domain V. These results suggested that the interaction of Domain V with hydrophobic compounds through the flexible loop is similar to the binding of hydrophobic compounds to the protein folding intermediate. PMID- 11434779 TI - Yeast ribosomal protein deletion mutants possess altered peptidyltransferase activity and different sensitivity to cycloheximide. AB - The major function of the ribosome is its ability to catalyze formation of peptide bonds, and it is carried out by the ribosomal peptidyltransferase. Recent evidence suggests that the catalyst of peptide bond formation is the 23S rRNA of the large ribosomal subunit. We have developed an in vitro system for the determination of peptidyltransferase activity in yeast ribosomes. Using this system, a kinetic analysis of a model reaction for peptidyltransferase is described with Ac-Phe-tRNA as the peptidyl donor and puromycin as the acceptor. The Ac-Phe-tRNA-poly(U)-80S ribosome complex (complex C) was isolated and then reacted with excess puromycin to give Ac-Phe-puromycin. This reaction (puromycin reaction) followed first-order kinetics. At saturating concentrations of puromycin, the first-order rate constant (k(3)) is identical to the catalytic rate constant (k(cat)) of peptidyltransferase. This k(cat) from wild-type yeast strains was equal to 2.18 min(-1) at 30 degrees C. We now present for the first time kinetic evidence that yeast ribosomes lacking a particular protein of the 60S subunit may possess significantly altered peptide bond-forming ability. The k(cat) of peptidyltransferase from mutants lacking ribosomal protein L24 was decreased 3-fold to 0.69 min(-1), whereas the k(cat) from mutants lacking L39 was slightly increased to 3.05 min(-1) and that from mutants lacking both proteins was 1.07 min(-1). These results suggest that the presence of ribosomal proteins L24 and, to a lesser extent, L39 is required for exhibition of the normal catalytic activity of the ribosome. Finally, the L24 or L39 mutants did not affect the rate or the extent of the translocation phase of protein synthesis. However, the absence of L24 caused increased resistance to cycloheximide, a translocation inhibitor. Translocation of Ac-Phe-tRNA from the A- to P-site was inhibited by 50% at 1.4 microM cycloheximide for the L24 mutant compared to 0.7 microM for the wild type. PMID- 11434780 TI - Ion concentration and temperature dependence of DNA binding: comparison of PurR and LacI repressor proteins. AB - Purine repressor (PurR) binding to specific DNA is enhanced by complexing with purines, whereas lactose repressor (LacI) binding is diminished by interaction with inducer sugars despite 30% identity in their protein sequences and highly homologous tertiary structures. Nonetheless, in switching from low- to high affinity DNA binding, these proteins undergo a similar structural change in which the hinge region connecting the DNA and effector binding domains folds into an alpha-helix and contacts the DNA minor groove. The differences in response to effector for these proteins should be manifest in the polyelectrolyte effect which arises from cations displaced from DNA by interaction with positively charged side chains on a protein and is quantitated by measurement of DNA binding affinity as a function of ion concentration. Consistent with structural data for these proteins, high-affinity operator DNA binding by the PurR-purine complex involved approximately 15 ion pairs, a value significantly greater than that for the corresponding state of LacI (approximately 6 ion pairs). For both proteins, however, conversion to the low-affinity state results in a decrease of approximately 2-fold in the number of cations released per dimeric DNA binding site. Heat capacity changes (DeltaC(p)) that accompany DNA binding, derived from buried apolar surface area, coupled folding, and restriction of motional freedom of polar groups in the interface, also reflect the differences between these homologous repressor proteins. DNA binding of the PurR-guanine complex is accompanied by a DeltaC(p) (-2.8 kcal mol(-1) K(-1)) more negative than that observed previously for LacI (-0.9 to -1.5 kcal mol(-1) K(-1)), suggesting that more extensive protein folding and/or enhanced structural rigidity may occur upon DNA binding for PurR compared to DNA binding for LacI. The differences between these proteins illustrate plasticity of function despite high-level sequence and structural homology and undermine efforts to predict protein behavior on the basis of such similarities. PMID- 11434781 TI - Misacylation and editing by Escherichia coli valyl-tRNA synthetase: evidence for two tRNA binding sites. AB - Valyl-tRNA synthetase (ValRS) has difficulty discriminating between its cognate amino acid, valine, and structurally similar amino acids. To minimize translational errors, the enzyme catalyzes a tRNA-dependent editing reaction that prevents accumulation of misacylated tRNA(Val). Editing occurs with threonine, alanine, serine, and cysteine, as well as with several nonprotein amino acids. The 3'-end of tRNA plays a vital role in promoting the tRNA-dependent editing reaction. Valine tRNA having the universally conserved 3'-terminal adenosine replaced by any other nucleoside does not stimulate the editing activity of ValRS. As a result 3'-end tRNA(Val) mutants, particularly those with 3'-terminal pyrimidines, are stably misacylated with threonine, alanine, serine, and cysteine. Valyl-tRNA synthetase is unable to hydrolytically deacylate misacylated tRNA(Val) terminating in 3'-pyrimidines but does deacylate mischarged tRNA(Val) terminating in adenosine or guanosine. Evidently, a purine at position 76 of tRNA(Val) is essential for translational editing by ValRS. We also observe misacylation of wild-type and 3'-end mutants of tRNA(Val) with isoleucine. Valyl tRNA synthetase does not edit wild-type tRNA(Val)(A76) mischarged with isoleucine, presumably because isoleucine is only poorly accommodated at the editing site of the enzyme. Misacylated mutant tRNAs as well as 3'-end-truncated tRNA(Val) are mixed noncompetitive inhibitors of the aminoacylation reaction, suggesting that ValRS, a monomeric enzyme, may bind more than one tRNA(Val) molecule. Gel-mobility-shift experiments to characterize the interaction of tRNA(Val) with the enzyme provide evidence for two tRNA binding sites on ValRS. PMID- 11434782 TI - Solid-state nuclear magnetic resonance evidence for an extended beta strand conformation of the membrane-bound HIV-1 fusion peptide. AB - Solid-state nuclear magnetic resonance (NMR) spectroscopy was applied to the membrane-bound form of a synthetic peptide representing the 23-residue N-terminal fusion peptide domain of the HIV-1 gp41 envelope glycoprotein. 1D solid-state NMR line width measurements of singly 13C carbonyl labeled peptides showed that a significant population of the membrane-bound peptide is well-structured in its N terminal and central regions while the C-terminus has more disorder. There was some dependence of line width on lipid composition, with narrower line widths and hence greater structural order observed for a lipid composition comparable to that found in the virus and its target T cells. In the more ordered N-terminal and central regions of the peptide, the 13C carbonyl chemical shifts are consistent with a nonhelical membrane-bound conformation. Additional evidence for a beta strand membrane-bound conformation was provided by analysis of 2D rotor synchronized magic angle spinning NMR spectra of doubly 13C carbonyl labeled peptides. Lipid mixing and aqueous contents leakage assays were applied to demonstrate the fusogenicity of the peptide under conditions comparable to those used for the solid-state NMR sample preparation. PMID- 11434783 TI - Analysis of sulfur biochemistry of sulfur bacteria using X-ray absorption spectroscopy. AB - Many sulfide-oxidizing organisms, including the photosynthetic sulfur bacteria, store sulfur in "sulfur globules" that are readily detected microscopically. The chemical form of sulfur in these globules is currently the focus of a debate, because they have been described as "liquid" by some observers, although no known allotrope of sulfur is liquid at physiological temperatures. In the present work we have used sulfur K-edge X-ray absorption spectroscopy to identify and quantify the chemical forms of sulfur in a variety of bacterial cells, including photosynthetic sulfur bacteria. We have also taken advantage of X-ray fluorescence self-absorption to derive estimates of the size and density of the sulfur globules in photosynthetic bacteria. We find that the form of sulfur that most resembles the globule sulfur is simply solid S(8), rather than more exotic forms previously proposed. PMID- 11434784 TI - Equilibrative nucleoside transporters: mapping regions of interaction for the substrate analogue nitrobenzylthioinosine (NBMPR) using rat chimeric proteins. AB - The rat equilibrative nucleoside transporters rENT1 and rENT2 belong to a family of integral membrane proteins with 11 potential transmembrane segments (TMs) and are distinguished functionally by differences in sensitivity to inhibition by nitrobenzylthioinosine (NBMPR). Structurally, the proteins have a large glycosylated extracellular loop between TMs 1 and 2 and a large cytoplasmic loop between TMs 6 and 7. In the present study, we have generated chimeras between NBMPR-sensitive rENT1 and NBMPR-insensitive rENT2, using splice sites at rENT1 residues 99 (end of TM 2), 171 (between TMs 4 and 5), and 231 (end of TM 6) to identify structural domains of rENT1 responsible for transport inhibition by NBMPR. Transplanting the amino-terminal half of rENT2 into rENT1 rendered rENT1 NBMPR-insensitive. Domain swaps within the amino-terminal halves of rENT1 and rENT2 identified two contiguous regions, TMs 3-4 (rENT1 residues 100-171) and TMs 5-6 (rENT1 residues 172-231), as the major sites of NBMPR interaction. Since NBMPR is a nucleoside analogue and functions as a competitive inhibitor of zero trans nucleoside influx, TMs 3-6 are likely to form parts of the substrate translocation channel. PMID- 11434785 TI - Ca2+-myristoyl switch and membrane binding of chemically acylated neurocalcins. AB - Neurocalcin is a member of a novel family of neuronal calcium sensors that belongs to the superfamily of EF-hand Ca(2+)-binding proteins. Neurocalcin is myristoylated on its N-terminus in vivo and can associate with biological membranes in a calcium and myristoyl-dependent manner. This process known as "Ca(2+)-myristoyl switch" has been best described for the photoreceptor specific protein, recoverin, as well as for several other neuronal calcium sensors. Here, we used reversed micelles to chemically acylate nonmyristoylated neurocalcin at its N-terminus with fatty acids of different lengths (from C12 to C16). This approach allowed us to prepare neurocalcin derivatives in which a single fatty acid is selectively linked to the N-terminal glycine of the polypeptide chain through an amide bond. The membrane binding properties of the monoacylated neurocalcins were then examined by cosedimentation with phospholipid vesicles and direct binding to lipid monolayers by surface plasmon resonance spectroscopy (Biacore). Our results show that neurocalcins monoacylated with lauric, myristic, or palmitic acid were able to associate with membrane in a calcium-dependent manner. This indicates that the Ca(2+)-myristoyl switch can function with different lipid moieties and is not strictly restricted to myristate. The ability to modify at will the fatty acid linked to the N-terminal glycine should be useful to analyze the contribution of the fatty acid moiety to the biological function of this family of neuronal calcium sensors. PMID- 11434786 TI - Role of disulfide bonds in the stability of recombinant manganese peroxidase. AB - Phanerochaete chrysosporium manganese peroxidase (MnP) [isoenzyme H4] was engineered with additional disulfide bonds to provide structural reinforcement to the proximal and distal calcium-binding sites. This rational protein engineering investigated the effects of multiple disulfide bonds on the stabilization of the enzyme heme environment and oxidase activity. Stabilization of the heme environment was monitored by UV-visible spectroscopy based on the electronic state of the alkaline transition species of ferric and ferrous enzyme. The optical spectral data confirm an alkaline transition to hexacoordinate, low-spin heme species for native and wild-type MnP and show that the location of the engineered disulfide bonds in the protein can have significant effects on the electronic state of the enzyme. The addition of a single disulfide bond in the distal region of MnP resulted in an enzyme that maintained a pentacoordinate, high-spin heme at pH 9.0, whereas MnP with multiple engineered disulfide bonds did not exhibit an increase in stability of the pentacoordinate, high-spin state of the enzyme at alkaline pH. The mutant enzymes were assessed for increased stability by incubation at high pH. In comparison to wild-type MnP, enzymes containing engineered disulfide bonds in the distal and proximal regions of the protein retained greater levels of activity when restored to physiological pH. Additionally, when assayed for oxidase activity at pH 9.0, proteins containing engineered disulfide bonds exhibited slower rates of inactivation than wild-type MnP. PMID- 11434787 TI - Kinetic characterization of the pH-dependent oligomerization of R67 dihydrofolate reductase. AB - Protein-protein recognition results from the assembly of complementary surfaces on two molecules that form a stable, noncovalent, specific complex. Our interest was to describe kinetic aspects of the recognition in order to understand the subtle molecular mechanism of association. R67 dihydrofolate reductase (DHFR) provides an ideal model to investigate kinetic parameters of protein-protein association since it is a homotetramer resulting from the pH-dependent dimerization of homodimers. We took advantage of the presence of a tryptophan residue at the dimer-dimer interface to monitor pH-dependent oligomerization of R67 DHFR using stopped-flow fluorescence techniques. Except for pH near neutrality where dissociation exhibited biphasic kinetics, association and dissociation followed monophasic kinetics fitted on a two-state model. Apparent rate constants of association k(on) and dissociation k(off) were determined at various pHs and pointed to the key role of a histidine located at the dimer-dimer interface in the pH control of tetramerization. The values of the tetramer-dimer equilibrium dissociation constant were calculated from the ratio k(off) /k(on) and correlated well with those previously measured at equilibrium. The thermodynamic parameters and the activation energies of both the association and dissociation were determined and indicated that the association is enthalpy driven and suggested that the formation of four hydrogen bonds (one per monomer) is responsible for the thermodynamic stability of the tetramer. Detailed analysis of the biphasic kinetics led to an original model, in which protonation of the tetramer is the triggering event for the dissociation process while the association involves primarily the unprotonated dimers. PMID- 11434789 TI - Impact of overweight on the risk of developing common chronic diseases during a 10-year period. AB - BACKGROUND: Overweight adults are at an increased risk of developing numerous chronic diseases. METHODS: Ten-year follow-up (1986-1996) of middle-aged women in the Nurses' Health Study and men in the Health Professionals Follow-up Study to assess the health risks associated with overweight. RESULTS: The risk of developing diabetes, gallstones, hypertension, heart disease, and stroke increased with severity of overweight among both women and men. Compared with their same-sex peers with a body mass index (BMI) (calculated as weight in kilograms divided by the square of height in meters) between 18.5 and 24.9, those with BMI of 35.0 or more were approximately 20 times more likely to develop diabetes (relative risk [RR], 17.0; 95% confidence interval [CI], 14.2-20.5 for women; RR, 23.4; 95% CI, 19.4-33.2 for men). Women who were overweight but not obese (ie, BMI between 25.0 and 29.9) were also significantly more likely than their leaner peers to develop gallstones (RR, 1.9), hypertension (RR, 1.7), high cholesterol level (RR, 1.1), and heart disease (RR, 1.4). The results were similar in men. CONCLUSIONS: During 10 years of follow-up, the incidence of diabetes, gallstones, hypertension, heart disease, colon cancer, and stroke (men only) increased with degree of overweight in both men and women. Adults who were overweight but not obese (ie, 25.0 < or = BMI < or = 29.9) were at significantly increased risk of developing numerous health conditions. Moreover, the dose response relationship between BMI and the risk of developing chronic diseases was evident even among adults in the upper half of the healthy weight range (ie, BMI of 22.0-24.9), suggesting that adults should try to maintain a BMI between 18.5 and 21.9 to minimize their risk of disease. PMID- 11434790 TI - Hyperhomocystinemia: a risk factor or a consequence of coronary heart disease? AB - BACKGROUND: Mild hyperhomocystinemia has been suggested as an indicator of an increased risk of cardiovascular disease. OBJECTIVE: To examine whether serum homocysteine concentration is a predictor of coronary heart disease (CHD) events. METHODS: A case-control study, nested in a population-based cohort study was used. During a follow-up of 13 years, 166 major coronary events (death from CHD or nonfatal myocardial infarction) occurred in men with evidence of heart disease at baseline and 272 events in men without a history of heart disease. Two controls per case were selected by individual matching. RESULTS: Among men with known heart disease at baseline, the relative risk (95% confidence interval) of CHD events adjusted for age, smoking, hypertension, diabetes mellitus, serum cholesterol level, body mass index, and alcohol consumption was 2.23 (95% confidence interval, 1.03-4.85) in the highest serum homocysteine quintile compared with the lowest quintile. Among the men free of heart disease at baseline, the corresponding relative risk was 0.90 (95% confidence interval, 0.51 1.60). CONCLUSIONS: This prospective study does not support the hypothesis that a high concentration of serum homocysteine is a risk factor for coronary events in a population free of heart disease. However, it does suggest that mild hyperhomocystinemia predicts secondary coronary events in men with heart disease, possibly as a consequence of atherosclerotic changes. PMID- 11434791 TI - Weight control in the physician's office. AB - BACKGROUND: Lifestyle changes involving diet, behavior, and physical activity are the cornerstone of successful weight control. Incorporating meal replacements (1 2 per day) into traditional lifestyle interventions may offer an additional strategy for overweight patients in the primary care setting. METHODS: One hundred thirteen overweight premenopausal women (mean +/- SD age, 40.4 +/- 5.5 years; weight, 82 +/- 10 kg; and body mass index, 30 +/- 3 kg/m(2)) participated in a 1-year weight-reduction study consisting of 26 sessions. The women were randomly assigned to 3 different traditional lifestyle-based groups: (1) dietitian-led group intervention (1 hour per session), (2) dietitian-led group intervention incorporating meal replacements (1 hour per session), or (3) primary care office intervention incorporating meal replacements with individual physician and nurse visits (10-15 minutes per visit). RESULTS: For the 74 subjects (65%) completing 1 year, the primary care office intervention using meal replacements was as effective as the traditional dietitian-led group intervention not using meal replacements (mean +/- SD weight loss, 4.3% +/- 6.5% vs 4.1% +/- 6.4%, respectively). Comparison of the dietitian-led groups showed that women using meal replacements maintained a significantly greater weight loss (9.1% +/- 8.9% vs 4.1% +/- 6.4%) (P =.03). Analysis across groups showed that weight loss of 5% to 10% was associated with significant (P =.01) reduction in percentage of body fat, body mass index, waist circumference, resting energy expenditure, insulin level, total cholesterol level, and low-density lipoprotein cholesterol level. Weight loss of 10% or greater was associated with additional significant (P =.05) improvements in blood pressure and triglyceride level. CONCLUSIONS: A traditional lifestyle intervention using meal replacements can be effective for weight control and reduction in risk of chronic disease in the physician's office setting as well as in the dietitian-led group setting. PMID- 11434792 TI - Body mass index and asthma in the military population of the northwestern United States. AB - BACKGROUND: Patients with asthma commonly have other medical problems such as obesity, but it is unclear if obesity independently relates to asthma occurrence. OBJECTIVE: To examine the association between asthma and obesity. METHODS: We studied enrollees aged 17 to 96 years in region 11 of TRICARE, a military managed health care program encompassing Washington, Oregon, and northern Idaho, using an enrollment questionnaire from January 1997 to December 1998. We performed case control analyses on 2788 asthma cases and 39 637 controls. From these cases and controls, we selected a random sample of 1000 asthma cases and 1000 controls, linking them to a computerized military health record system to verify if medications indicated for asthma therapy were prescribed. After excluding cases not prescribed bronchodilator medications and excluding controls prescribed bronchodilator medications or steroids, we used logistic regression to estimate associations among asthma, body mass index, and demographic, lifestyle, and comorbid risk factors in 386 verified cases and 744 verified controls. RESULTS: Increasing body mass index, younger age, female sex, non-active duty beneficiary status, and arthritis were significant independent predictors of asthma prevalence in both our larger analysis and our verified substudy, whereas stomach ulcer, depression, hypertension, and white race are also independent predictors of asthma prevalence in our larger analysis. CONCLUSIONS: Increasing body mass index is a key factor predicting prevalence of asthma and, if determined to be etiologically related to asthma incidence, is a potentially modifiable risk factor for asthma. PMID- 11434793 TI - Cyclobenzaprine and back pain: a meta-analysis. AB - BACKGROUND: Back pain is a common problem for which cyclobenzaprine hydrochloride is frequently prescribed. OBJECTIVE: To perform a systematic review of cyclobenzaprine's effectiveness in the treatment of back pain. METHODS: We searched MEDLINE, PsycLIT, CINAHL, EMBASE, AIDSLINE, HEALTHSTAR, CANCERLIT, the Cochrane Library, Micromedex, Federal Research in Progress, and the references of reviewed articles, and contacted Merck, Sharpe and Dohme for English-language, randomized, placebo-controlled trials of cyclobenzaprine in adults with back pain. Outcomes included global improvement and 5 specific domains of back pain (local pain, muscle spasm, range of motion, tenderness to palpation, and activities of daily living). Study quality was assessed using the methods of Jadad. Summary outcomes were obtained using a random-effects model. RESULTS: Patients treated with cyclobenzaprine were nearly 5 times (odds ratio, 4.7; 95% confidence interval, 2.7-8.1) as likely to report symptom improvement by day 14 as were those treated with placebo. Slightly fewer than 3 individuals (2.7; 95% confidence interval, 2.0-4.2) needed treatment for 1 to improve. The magnitude of this improvement was modest, with an effect size of 0.38 to 0.58 in all 5 outcomes (local pain, muscle spasm, tenderness to palpation, range of motion, and activities of daily living). Treatment efficacy for these 5 outcomes was greatest early, in the first few days of treatment, declining after the first week. Patients receiving cyclobenzaprine also experienced more adverse effects, the most common being drowsiness. CONCLUSIONS: Cyclobenzaprine is more effective than placebo in the management of back pain; the effect is modest and comes at the price of greater adverse effects. The effect is greatest in the first 4 days of treatment, suggesting that shorter courses may be better. Studies comparing the relative value of acetaminophen, nonsteroidal anti-inflammatory drugs, and cyclobenzaprine individually and in combination in the treatment of back pain are needed. PMID- 11434794 TI - Ineffective secondary prevention in survivors of cardiovascular events in the US population: report from the Third National Health and Nutrition Examination Survey. AB - BACKGROUND: Survivors of myocardial infarction (MI) or stroke are at high risk for subsequent cardiovascular events. There is limited assessment of the effectiveness of risk factor modification through current secondary preventive strategies in the US population. We determined the adequacy of risk factor modification in 1252 survivors of MI, stroke, or both in a nationally representative sample of US adults and identified factors related to inadequate control of risk factors. METHODS: The adequacy of control for hypertension, diabetes mellitus, cigarette smoking, alcohol use, and hypercholesterolemia was assessed by personal interview, blood pressure measurements, and serum glycosylated hemoglobin and cholesterol levels in 17 752 US adults who participated in the Third National Health and Nutrition Examination Survey between 1988 and 1994. We also evaluated the role of potentially related factors, including age, sex, race/ethnicity, educational attainment, socioeconomic status, and medical insurance status using multivariate logistic regression analysis. RESULTS: Of 738 known hypertensive persons, hypertension was uncontrolled in 388 (53%). Previously undiagnosed hypertension was detected in 138 others (11%). Of 289 diabetic persons, serum glucose control was inadequate in 141. Of 1252 survivors, 225 (18%) were currently smoking, and heavy alcohol use was observed in 56 persons. Hypercholesterolemia was poorly controlled in 185 (46%) of 405 persons with known hypercholesterolemia. Undetected hypercholesterolemia was observed in 160 persons (13%). In the multivariate analysis, high-risk profiles were more likely to be observed in persons aged 46 to 65 years, women, and African Americans. CONCLUSIONS: High prevalence of inadequate secondary prevention was found in a subset of the US population at highest risk for stroke and MI. Considerable efforts are required to effectively implement risk factor modification strategies after MI or stroke, particularly in middle-aged persons, African Americans, and women. PMID- 11434795 TI - Risk factors for adverse drug events among nursing home residents. AB - BACKGROUND: In a prospective study of nursing home residents, we found adverse drug events (ADEs) to be common, serious, and often preventable. To direct prevention efforts at high-risk residents, information is needed on resident level risk factors. METHODS: Case-control study nested within a prospective study of ADEs among residents in 18 nursing homes. For each ADE, we randomly selected a control from the same home. Data were abstracted from medical records on functional status, medical conditions, and medication use. RESULTS: Adverse drug events were identified in 410 nursing home residents. Independent risk factors included being a new resident (odds ratio [OR], 2.8; 95% confidence interval [CI], 1.5-5.2) and taking anti-infective medications (OR, 4.0; CI, 2.5-6.2), antipsychotics (OR, 3.2; CI, 2.1-4.9), or antidepressants (OR, 1.5; CI, 1.1-2.3). The number of regularly scheduled medications was associated with increased risk of ADEs; the OR associated with taking 5 to 6 medications was 2.0 (CI, 1.2-3.2); 7 to 8 medications, 2.8 (CI, 1.7-4.7); and 9 or more, 3.3 (CI, 1.9-5.6). Taking supplements or nutrients was associated with lower risk (OR, 0.42; CI, 0.27 0.63). Preventable ADEs occurred in 226 residents. Independent risk factors included taking opioid medications (OR, 6.6; CI, 2.3-19.3), antipsychotics (OR, 4.0; CI, 2.2-7.3), anti-infectives (OR, 3.0; CI, 1.6-5.8), antiepileptics (OR, 2.2; CI, 1.1-4.5), or antidepressants (OR, 2.0; CI, 1.1-3.5). Scores of 5 or higher on the Charlson Comorbidity Index were associated with increased risk of ADEs (OR, 2.6; CI, 1.1-6.0). The number of regularly scheduled medications was also a risk factor: the OR for 7 to 8 medications was 3.2 (CI, 1.4-6.9) and for 9 or more, 2.9 (CI, 1.3-6.8). Residents taking nutrients or supplements were at lower risk (OR, 0.27; CI, 0.14-0.50). CONCLUSIONS: It is possible to identify nursing home residents at high risk of having an ADE. Particular attention should be directed at new residents, those with multiple medical conditions, those taking multiple medications, and those taking psychoactive medications, opioids, or anti-infective drugs. PMID- 11434796 TI - Women's interest in chemoprevention for breast cancer. AB - BACKGROUND: Chemoprevention is the use of pharmacologic or natural agents to inhibit the development of cancer. Tamoxifen citrate is the only approved chemopreventive agent for breast cancer. We sought to determine whether women are interested in taking a drug to prevent breast cancer and to assess the relationship between objective and subjective breast cancer risk and interest in chemoprevention. METHODS: We conducted telephone interviews (November 3, 1997, to May 6, 1998) among a community sample of women aged 40 to 45 and 50 to 55 years enrolled in a randomized controlled trial to evaluate the efficacy of a tailored mammography decision aid. Objective breast cancer risk was measured using the 5 year Gail score. Subjective breast cancer risk was measured using perceptions of absolute risk, perceptions of comparative risk, and worry about getting breast cancer. At 12-month follow-up (November 2, 1998, to July 20, 1999), we measured interest in taking a drug to prevent breast cancer. RESULTS: Among the 1273 women surveyed, 23% were interested in taking a drug to prevent breast cancer; 8% were potentially eligible for tamoxifen therapy (5-year Gail score > or = 1.66%). Eligibility for chemoprevention, based on the 5-year Gail score, was not associated with interest in taking a drug to prevent breast cancer. Women who were worried about breast cancer were 3 times more likely to be interested in taking a drug to prevent breast cancer than those who were not worried. CONCLUSION: Women's interest in chemoprevention might arise more from worries about getting breast cancer than from their objective risk factors. PMID- 11434797 TI - Ten years of life: Is it a matter of choice? AB - BACKGROUND: Relative risk estimates suggest that effective implementation of behaviors commonly advocated in preventive medicine should increase life expectancy, although there is little direct evidence. OBJECTIVE: To test the hypothesis that choices regarding diet, exercise, and smoking influence life expectancy. METHODS: A total of 34 192 California Seventh-Day Adventists (75% of those eligible) were enrolled in a cohort and followed up from 1976 to 1988. A mailed questionnaire provided dietary and other exposure information at study baseline. Mortality for all subjects was ascertained by matching to state death tapes and the National Death Index. RESULTS: California Adventists have higher life expectancies at the age of 30 years than other white Californians by 7.28 years (95% confidence interval, 6.59-7.97 years) in men and by 4.42 years (95% confidence interval, 3.96-4.88 years) in women, giving them perhaps the highest life expectancy of any formally described population. Commonly observed combinations of diet, exercise, body mass index, past smoking habits, and hormone replacement therapy (in women) can account for differences of up to 10 years of life expectancy among Adventists. A comparison of life expectancy when these factors take high-risk compared with low-risk values shows independent effects that vary between 1.06 and 2.74 years for different variables. The effect of each variable is assessed with all others at either medium- or high-risk levels. CONCLUSIONS: Choices regarding diet, exercise, cigarette smoking, body weight, and hormone replacement therapy, in combination, appear to change life expectancy by many years. The longevity experience of Adventists probably demonstrates the beneficial effects of more optimal behaviors. PMID- 11434798 TI - Hypoglycemia in patients with type 2 diabetes mellitus. AB - BACKGROUND: Although hypoglycemia is the most common complication of intensive diabetes therapy, there is little information about risk factors for hypoglycemia in patients with type 2 diabetes mellitus. OBJECTIVE: To determine the prevalence and predisposing factors for hypoglycemia in patients with type 2 diabetes. METHODS: Retrospective, cross-sectional analysis set in an outpatient specialty diabetes clinic. We included those patients who had baseline and follow-up visits from April 1 through October 31, 1999. Hypoglycemia was defined as typical symptoms relieved by eating, and/or blood glucose level of less than 60 mg/dL (<3.3 mmol/L). Univariate and multivariate logistic regression were used to determine the contributions to hypoglycemia of age, sex, diabetes duration, body mass index (calculated as weight in kilograms divided by the square of height in meters), fasting plasma glucose level, glycosylated hemoglobin (HbA(1c)) level, type of therapy, and previous episodes at the follow-up visit. RESULTS: We studied 1055 patients. Prevalence of hypoglycemic symptoms was 12% (9/76) for patients treated with diet alone, 16% (56/346) for those using oral agents alone, and 30% (193/633) for those using any insulin (P<.001). Severe hypoglycemia occurred in only 5 patients (0.5%), all using insulin. Multiple logistic regression analysis demonstrated that insulin therapy, lower HbA(1c) level at follow-up, younger age, and report of hypoglycemia at the baseline visit were independently associated with increased prevalence of hypoglycemia. There were no significant predictors of severe hypoglycemia. CONCLUSIONS: Mild hypoglycemia is common in patients with type 2 diabetes undergoing aggressive diabetes management, but severe hypoglycemia is rare. Concerns about hypoglycemia should not deter efforts to achieve tight glycemic control in most patients with type 2 diabetes. PMID- 11434799 TI - Race and sex differences in consistency of care with national asthma guidelines in managed care organizations. AB - BACKGROUND: In the United States, morbidity from asthma disproportionately affects African Americans and women. Although inadequate care contributes to overall asthma morbidity, less is known about differences in asthma care by race and sex. SUBJECTS AND METHODS: To examine the relationships of race and sex with asthma care, we analyzed responses to questionnaires administered to adults enrolled in 16 managed care organizations participating in the Outcomes Management System Asthma Study between September and December 1993. Indicators of care consistent with National Asthma Education and Prevention Program (1991) recommendations were assessed. Of a random sample of 8640 patients asked to participate, 6612 (77%) completed the survey. This study focused on 5062 (14% African American, 72% women) patients with at least moderate asthma symptom severity. RESULTS: Fewer African Americans than whites reported care consistent with recommendations for medication use (eg, daily inhaled corticosteroid use, 34.9% vs 54.4%; P =.001), self-management education (eg, action plan, 42.0% vs 53.8%; P =.001), avoiding triggers (37.6% vs 53.6%; P =.001), and specialist care (28.3% vs 41.0%; P =.001). Differences in asthma care by sex were smaller and tended to favor women except for daily inhaled corticosteroid use (women vs men: 49.6% vs 58.3%; P =.001) and having specialist care (37.7% vs 43.1%; P =.001). Similar race and sex differences were observed after adjusting for age, education, employment, and symptom frequency. CONCLUSIONS: Even among patients with health insurance, disparities in asthma care for African Americans compared with whites exist and may contribute to race disparities in outcomes. Women generally reported better asthma care but may benefit from greater use of inhaled corticosteroids. PMID- 11434800 TI - Association between peak expiratory flow and the development of carotid atherosclerotic plaques. AB - BACKGROUND: Numerous population-based studies have suggested that impaired lung function is associated with subsequent coronary heart diseases-related mortality and cardiovascular disease-related mortality. The relative contribution of atherosclerosis in these associations is unknown. OBJECTIVE: To examine the association of peak expiratory flow (PEF) with the occurrence during 4 years of atherosclerotic plaques in the extracranial carotid arteries in a sample of 656 subjects (aged 59-71 years) free of coronary heart disease and stroke at baseline. METHODS: Peak expiratory flow was measured at the baseline examination. Peak expiratory flow values relative to the predicted values (relative PEF values) were calculated, predicted values being obtained from previously published sex-specific regression equations of PEF on age and height. A carotid B mode ultrasonographic examination was performed at baseline and 2 and 4 years later. The occurrence of carotid plaques during follow-up was defined as the appearance of 1 plaque (or more) in previously normal carotid segments and/or the appearance of new plaques in the carotid segments that previously had plaques. RESULTS: The proportion of subjects who experienced an occurrence of carotid atherosclerotic plaques during follow-up was 16.8% (110/656). The unadjusted odds ratios from the highest to the lowest quintiles of relative PEF values were 1.00, 1.07 (95% confidence interval [CI], 0.69-2.79), 1.08 (95% CI, 0.52-2.24), 1.38 (95% CI, 0.69-2.79), and 3.07 (95% CI, 1.62-5.85) (P<.001 for trend). Adjustment for major known cardiovascular risk factors did not markedly change the results, and the multivariate-adjusted odds ratio of carotid plaque occurrence in subjects with the lowest quintile of PEF compared with those with the highest quintile remained highly significant (odds ratio, 2.84; 95% CI, 1.45-5.71) (P =.002). Particularly in all smoking categories, carotid plaque occurrence was higher in subjects with the lowest relative PEF values. In never smokers, the multivariate adjusted odds ratio of carotid plaque occurrence in subjects with the lowest quintile of PEF compared with those with the highest quintile was 2.80 (95% CI, 1.14-6.88). CONCLUSIONS: Reduced lung function predicts the development of carotid atherosclerosis in elderly subjects. The nature of these associations remains largely unknown and merits further investigations. Nevertheless, assessment of lung function, which is simple and inexpensive, could help identify a population at high risk of atherosclerosis development and coronary heart disease. PMID- 11434801 TI - Acute fulminant hepatitis after treatment with rabeprazole and terbinafine. PMID- 11434802 TI - Hypercholesterolemia as a possible risk factor for osteopenia in type 2 diabetes mellitus. PMID- 11434803 TI - Integrating complementary and alternative medicine into conventional medical education: role of basic science. PMID- 11434804 TI - Awareness of CAM among residents: a long way to go. PMID- 11434805 TI - Possible danger of integrating CAM in medical education. PMID- 11434807 TI - Quinolones for community-acquired pneumonia. PMID- 11434810 TI - A piece of my mind. Brave, waiting for Pasteur. PMID- 11434811 TI - Global issues on the agenda at the World Health Assembly: discussion of HIV/AIDS, leprosy, access to drugs. PMID- 11434812 TI - Seeding the world with stem cells. PMID- 11434813 TI - Experimental prostate cancer drugs slow disease progression. PMID- 11434814 TI - From the Food and Drug Administration. PMID- 11434815 TI - St John's wort and major depression. PMID- 11434816 TI - St John's wort and major depression. PMID- 11434817 TI - St John's wort and major depression. PMID- 11434818 TI - St john's wort and major depression. PMID- 11434819 TI - St John's wort and major depression. PMID- 11434821 TI - Prenatal cocaine exposure as a risk factor for later developmental outcomes. PMID- 11434822 TI - Prenatal cocaine exposure as a risk factor for later developmental outcomes. PMID- 11434823 TI - Prenatal cocaine exposure as a risk factor for later developmental outcomes. PMID- 11434825 TI - Informed consent for public automated external defibrillation. PMID- 11434826 TI - Effect of short-course, high-dose amoxicillin therapy on resistant pneumococcal carriage: a randomized trial. AB - CONTEXT: Emerging drug resistance threatens the effectiveness of existing therapies for pneumococcal infections. Modifying the dose and duration of antibiotic therapy may limit the spread of resistant pneumococci. OBJECTIVE: To determine whether short-course, high-dose amoxicillin therapy reduces risk of posttreatment resistant pneumococcal carriage among children with respiratory tract infections. DESIGN AND SETTING: Randomized trial conducted in an outpatient clinic in Santo Domingo, Dominican Republic, October 1999 through July 2000. PARTICIPANTS: Children aged 6 to 59 months who were receiving antibiotic prescriptions for respiratory tract illness (n = 795). INTERVENTIONS: Children were randomly assigned to receive 1 of 2 twice-daily regimens of amoxicillin: 90 mg/kg per day for 5 days (n = 398) or 40 mg/kg per day for 10 days (n = 397). MAIN OUTCOME MEASURES: Penicillin-nonsusceptible Streptococcus pneumoniae carriage, assessed in nasopharyngeal specimens collected at days 0, 5, 10, and 28; baseline risk factors for nonsusceptible pneumococcal carriage; and adherence to regimen, compared between the 2 groups. RESULTS: At the day 28 visit, risk of penicillin-nonsusceptible pneumococcal carriage was significantly lower in the short-course, high-dose group (24%) compared with the standard-course group (32%); relative risk (RR), 0.77; 95% confidence interval (CI), 0.60-0.97; P =.03; risk of trimethoprim-sulfamethoxazole nonsusceptibility was also lower in the short-course, high-dose group (RR, 0.77; 95% CI, 0.58-1.03; P =.08). The protective effect of short-course, high-dose therapy was stronger in households with 3 or more children (RR, 0.72; 95% CI, 0.52-0.98). Adherence to treatment was higher in the short-course, high-dose group (82% vs 74%; P =.02). CONCLUSION: Short-course, high-dose outpatient antibiotic therapy appears promising as an intervention to minimize the impact of antibiotic use on the spread of drug resistant pneumococci. PMID- 11434827 TI - Evaluation of decision rules for referring women for bone densitometry by dual energy x-ray absorptiometry. AB - CONTEXT: Identification of women with low bone mineral density (BMD) is an important strategy in reducing the incidence of osteoporotic fractures. However, screening all women is not recommended. OBJECTIVES: To assess the diagnostic properties of 4 decision rules--Simple Calculated Osteoporosis Risk Estimation (SCORE), Osteoporosis Risk Assessment Instrument (ORAI), Age, Body Size, No Estrogen (ABONE), and body weight less than 70 kg (weight criterion)--for selecting women for dual-energy x-ray absorptiometry (DXA) testing and to compare results with recommendations made in the National Osteoporosis Foundation (NOF) practice guidelines. DESIGN AND SETTING: Analysis of data from the Canadian Multicentre Osteoporosis Study, a population-based community sample, collected from 9 study centers across Canada between February 1996 and September 1997. PARTICIPANTS: Postmenopausal women aged 45 years or older (N = 2365) without bone disease who had DXA data for the femoral neck, data to apply selection criteria, and who were not currently taking estrogens or who had been taking hormone replacement therapy for 5 or more years. MAIN OUTCOME MEASURES: Sensitivity, specificity, and area under the receiver operating characteristic (AUROC) curve of each of the 4 decision rules and the NOF guidelines for identifying women with a BMD T score of less than -1.0 SD, less than -2.0 SD, and no more than -2.5 SD at the femoral neck, and percentages of women recommended for testing, stratified by BMD level and age. RESULTS: The percent of women with a BMD T score less than 1, less than -2, and no more than -2.5 were 68.3%, 25.4%, and 10.0%, respectively. The AUROC curves were greatest using SCORE and ORAI. The sensitivity for identifying women with a BMD T score of less than -2.0 was 93.7% (95% confidence interval [CI], 91.8%-95.6%) using the NOF guidelines and was 97.5% (95% CI, 96.3%-98.8%), 94.2% (95% CI, 92.3%-96.1%), 79.1% (95% CI, 75.9% 82.3%), and 79.6% (95% CI, 76.4%-82.8%), respectively, using the SCORE, ORAI, ABONE, and weight criterion. However, the NOF guidelines also resulted in 74.4% (95% CI, 71.3%-77.6%) of women with a normal BMD (T score of -1.0 or higher) being tested compared with 69.2% (95% CI, 65.9%-72.5%), 56.3% (95% CI, 52.7% 59.8%), 35.8% (95% CI, 32.4%-39.2%), and 38.1% (95% CI, 34.6%-41.6%), respectively, using the 4 decision rules. Assessments suggest that ABONE and weight criterion are not useful case-finding approaches. CONCLUSION: The SCORE and ORAI decision rules are better than the NOF guidelines at targeting BMD testing in high-risk patients. The acceptability of these rules in clinical practice merits further investigation given their potential effect on the use of densitometry services. PMID- 11434828 TI - Effect of statin therapy on C-reactive protein levels: the pravastatin inflammation/CRP evaluation (PRINCE): a randomized trial and cohort study. AB - CONTEXT: Plasma levels of the inflammatory biomarker C-reactive protein (CRP) predict cardiovascular risk, and retrospective studies suggest that 3-hydroxy-3 methylglutaryl coenzyme A reductase inhibitors (statins) may lower CRP in a manner largely independent of low-density lipoprotein cholesterol (LDL-C). However, prospective trial data directly evaluating this anti-inflammatory effect of statins are not available. OBJECTIVE: To test the hypothesis that pravastatin has anti-inflammatory effects as evidenced by CRP reduction. DESIGN, SETTING, AND PARTICIPANTS: Community-based, prospective, randomized, double-blind trial including 1702 men and women with no prior history of cardiovascular disease (primary prevention cohort) and open-label study including 1182 patients with known cardiovascular disease (secondary prevention cohort) who provided at least baseline and 12-week blood samples. The study was conducted in US office-based practices from February to December 2000. INTERVENTIONS: Participants in the double-blind primary prevention trial were randomly assigned to receive 40 mg/d of pravastatin (n = 865) or placebo (n = 837) for 24 weeks. Participants in the secondary prevention cohort received 40 mg/d of open-label pravastatin for 24 weeks. MAIN OUTCOME MEASURE: Change in CRP levels from baseline to 24 weeks. RESULTS: In the primary prevention trial, compared with placebo, pravastatin reduced median CRP levels by 16.9% (P<.001) at 24 weeks, reflecting a decrease of 0.02 mg/dL in the pravastatin group while no change in CRP levels was observed in the placebo group. This effect was seen as early as 12 weeks (median reduction in CRP with pravastatin, 14.7%; P<.001) and was present among all prespecified subgroups according to sex, age, smoking status, body mass index, baseline lipid levels, presence of diabetes, and use of aspirin or hormone replacement therapy. No significant association was observed between baseline CRP and baseline LDL-C levels, end-of-study CRP and end-of-study LDL-C levels, or change in CRP and change in LDL-C levels over time. In linear regression analyses, the only significant predictors of change in CRP on a log scale were randomized pravastatin allocation and baseline CRP levels (P<.001 for both). Similar reductions in CRP levels were observed at 12 weeks (-14.3%) and 24 weeks (-13.1%) in the secondary prevention cohort treated with pravastatin (P<.005 for both). CONCLUSIONS: In this prospective trial, pravastatin reduced CRP levels at both 12 and 24 weeks in a largely LDL-C-independent manner. These data provide evidence that statins may have anti-inflammatory effects in addition to lipid-lowering effects. PMID- 11434829 TI - Factors influencing families' consent for donation of solid organs for transplantation. AB - CONTEXT: Transplantation has become the therapy of choice for patients with organ failure. However, the low rate of consent by families of donor-eligible patients is a major limiting factor in the success of organ transplantation. OBJECTIVE: To explore factors associated with the decision to donate among families of potential solid organ donors. DESIGN AND SETTING: Data collection via chart reviews, telephone interviews with health care practitioners (HCPs) or organ procurement organization (OPO) staff, and face-to-face interviews with family for all donor-eligible deaths at 9 trauma hospitals in southwestern Pennsylvania and northeastern Ohio from 1994 to 1999. PARTICIPANTS: Family members, HCPs, and OPO staff involved in the donation decision for 420 donor-eligible patients. MAIN OUTCOME MEASURE: Factors associated with family decision to donate or not donate organs for transplantation. RESULTS: A total of 238 of the 420 cases led to organ donation; 182 did not. Univariate analysis revealed numerous factors associated with the donation decision. Multivariable analysis of associated variables revealed that family and patient sociodemographics (ethnicity, patient's age and cause of death) and prior knowledge of the patients' wishes were significantly associated with willingness to donate (adjusted odds ratio [OR], 7.68; 95% confidence interval [CI], 6.55-9.01). Families who discussed more topics and had more conversations about organ donation were more likely to donate (adjusted OR, 5.22; 95% CI, 4.32-6.30), as were families with more contact with OPO staff (adjusted OR, 3.08; 95% CI, 2.63-3.60) and those who experienced an optimal request pattern (adjusted OR, 2.96; 95% CI, 2.58-3.40). Socioemotional and communication variables acted as intervening variables. CONCLUSIONS: Public education is needed to modify attitudes about organ donation prior to a donation opportunity. Specific steps can be taken by HCPs and OPO staff to maximize the opportunity to persuade families to donate their relatives' organs. PMID- 11434830 TI - Risk of stroke associated with abciximab among patients undergoing percutaneous coronary intervention. AB - CONTEXT: Abciximab, a potent inhibitor of the platelet glycoprotein IIb/IIIa receptor, reduces thrombotic complications in patients undergoing percutaneous coronary intervention (PCI). Because of its potent inhibition of platelet aggregation, the effect of abciximab on risk of stroke is a concern. OBJECTIVE: To determine whether abciximab use among patients undergoing PCI is associated with an increased risk of stroke. DESIGN: Combined analysis of data from 4 double blind, placebo-controlled, randomized trials (EPIC, CAPTURE, EPILOG, and EPISTENT) conducted between November 1991 and October 1997 at a total of 257 academic and community hospitals in the United States and Europe. PATIENTS: A total of 8555 patients undergoing PCI with or without stent deployment for a variety of indications were randomly assigned to receive a bolus and infusion of abciximab (n = 5476) or matching placebo (n = 3079). One treatment group in EPIC received a bolus of abciximab only. MAIN OUTCOME MEASURE: Risk of hemorrhagic and nonhemorrhagic stroke within 30 days of treatment among abciximab and placebo groups. RESULTS: No significant difference in stroke rate was observed between patients assigned abciximab (n = 22 [0.40%]) and those assigned placebo (n = 9 [0.29%]; P =.46). Excluding the EPIC abciximab bolus-only group, there were 9 strokes (0.30%) among 3023 patients who received placebo and 15 (0.32%) in 4680 patients treated with abciximab bolus plus infusion, a difference of 0.02% (95% confidence interval [CI], -0.23% to 0.28%). The rate of nonhemorrhagic stroke was 0.17% in patients treated with abciximab and 0.20% in patients treated with placebo (difference, -0.03%; 95% CI, -0.23% to 0.17%), and the rates of hemorrhagic stroke were 0.15% and 0.10%, respectively (difference, 0.05%; 95% CI, -0.11% to 0.21%). Among patients treated with abciximab, the rate of hemorrhagic stroke in patients receiving standard-dose heparin in EPIC, CAPTURE, and EPILOG was higher than in those receiving low-dose heparin in the EPILOG and EPISTENT trials (0.27% vs 0.04%; P =.057). CONCLUSIONS: Abciximab in addition to aspirin and heparin does not increase the risk of stroke in patients undergoing PCI. Patients undergoing PCI and treated with abciximab should receive low-dose, weight-adjusted heparin. PMID- 11434831 TI - An analysis of physician antitrust exemption legislation: adjusting the balance of power. AB - Current antitrust law restricts physicians from joining together to collectively negotiate. However, such activities may be approved by state laws under the so called state action immunity doctrine and by federal legislation under an explicit antitrust exemption. In 1999, Texas became the first state to pass physician antitrust exemption legislation allowing physicians, under certain defined circumstances, to collectively negotiate fees with health plans. Last year, similar legislation was introduced in the US Congress, in 18 state legislatures, and in the District of Columbia. This legislation was passed only in the District of Columbia where its implementation was blocked by the city's financial control board. Nonetheless, legislation permitting physicians to collectively negotiate fees with managed care plans has been introduced in 10 state legislatures this year, and there is continued interest in introducing similar legislation in the US Congress. This analysis examines the basic features of this legislation and its potential impact on the balance of power between physicians and managed care plans. PMID- 11434832 TI - Reporting financial conflicts of interest and relationships between investigators and research sponsors. PMID- 11434833 TI - Placing PRINCE in perspective. PMID- 11434839 TI - Improving human immunodeficiency virus testing for adolescents. PMID- 11434840 TI - To help or not to help... That is the question. PMID- 11434841 TI - Maternal and paternal recreational drug use and sudden infant death syndrome. AB - OBJECTIVE: To determine whether maternal or paternal use of cocaine, opiates, or marijuana during conception and pregnancy and postnatally increases the risk of sudden infant death syndrome (SIDS) during the first year of the infant's life. This is an important issue and may prove useful in further decreasing the rate of SIDS. METHODS: A case-control study was conducted consisting of 239 infants who died of SIDS in southern California between 1989 and 1992, and 239 healthy infants who were matched on the basis of birth hospital, date of birth, age, and sex. Specific drug use at the period of conception, during pregnancy and breastfeeding, and in the presence or vicinity of the infant was ascertained by telephone for the white, African American, Hispanic, Asian American, and Pacific Islander case and control fathers and mothers. RESULTS: Maternal recreational drug use during pregnancy was not associated with the risk of SIDS after adjusting for maternal smoking during pregnancy (adjusted odds ratio [OR] = 2.0; 95% confidence interval [CI], 0.6-6.5). There were statistically significant differences between case and control fathers' use of marijuana during conception (OR = 2.2; 95% CI, 1.2-4.2; P =.01), during pregnancy (OR = 2.0; 95% CI, 1.0-4.1; P =.05), and postnatally (OR = 2.8; 95% CI, 1.1-7.3; P =.04) and the risk of SIDS, while adjusting for paternal smoking and alcohol use. CONCLUSIONS: There was no association between maternal recreational drug use and SIDS. Paternal marijuana use during the periods of conception and pregnancy and postnatally were significantly associated with SIDS. The role of paternal psychoactive drug use, especially the relationship between marijuana and SIDS, is an understudied area; however, before any definitive role for the father can be confirmed, these findings should be investigated and replicated in future studies. PMID- 11434842 TI - Maternal outcomes of a randomized controlled trial of a community-based support program for families of children with chronic illnesses. AB - BACKGROUND: Parents of children with chronic illnesses are at high risk for secondary mental health problems, such as anxiety and depression. OBJECTIVE: To evaluate maternal outcomes of a support intervention for families of children with selected chronic illnesses. DESIGN: A randomized controlled clinical trial design with repeated measures 1 year apart. SETTING: A community-based family support intervention linked to subspecialty and general pediatric clinics and practices in a metropolitan area. PARTICIPANTS: A population-based sample of 193 mothers of children aged 7 to 11 years; the children were diagnosed as having diabetes, sickle cell anemia, cystic fibrosis, or moderate to severe asthma. About 15% of the persons contacted refused to participate in the research, and 14% of the families were lost to follow-up. INTERVENTION: The 15-month intervention, the Family-to-Family Network, was designed to enhance mothers' mental health by linking mothers of school-aged children with selected chronic illnesses with mothers of older children with the same condition. The program included telephone contacts, face-to-face visits, and special family events. MAIN OUTCOME MEASURES: Beck Depression Inventory score and the Psychiatric Symptom Index. RESULTS: Maternal anxiety scores for participants in the experimental group decreased during the intervention period for all diagnostic groups and for the total group; scores for the control group increased (F = 5.07, P =.03). In multiple regression analyses, the intervention group was a significant predictor of posttest anxiety scores (P =.03). Effects were greater for mothers with high baseline anxiety (P<.001) and for those who were themselves in poor health (P<.01). CONCLUSIONS: A family support intervention can have beneficial effects on the mental health status of mothers of children with chronic illnesses. This type of intervention can be implemented in diverse pediatric settings. PMID- 11434843 TI - Langerhans cell histiocytosis presenting in the neonatal period: a retrospective case series. AB - OBJECTIVES: To describe the morphologic characteristics of skin lesions, extent of extracutaneous disease, and outcomes in patients with neonatal presentation of Langerhans cell histiocytosis (LCH), and to examine clinical predictors of disease prognosis. DESIGN: Retrospective validation cohort study. Maximum duration of follow-up was 10 years. SETTING: A tertiary care children's hospital in Chicago, Ill. PATIENTS: Nineteen children with cutaneous findings in the first 4 weeks of life and subsequently diagnosed with LCH based on compatible tissue histologic analysis, confirmed by electron microscopy and/or immunohistochemical analysis. MAIN OUTCOME MEASURE: Cutaneous lesion morphologic characteristics, extracutaneous manifestations, treatments, and outcomes were tabulated and compared. RESULTS: The most common initial skin lesion was erythematous, often crusted, vesiculopustules. Skin lesion morphologic traits did not correlate with extent of extracutaneous disease. One third of patients had disease limited to the skin and/or mucous membranes. All of these patients are alive and well, and 1 has developed diabetes insipidus. Twelve of the 19 patients had multisystem disease, and 2 died of disease. The results of a multiorgan workup performed at the time of diagnosis were predictive of which patients in this cohort manifested multisystem disease. The overall incidence of diabetes insipidus was 21%. CONCLUSIONS: Vesiculopustular lesions are common in congenital/neonatal LCH, but the morphologic characteristics of lesions are not helpful in predicting the extent of disease. A multiorgan evaluation at the time of diagnosis may be predictive of the probability of multisystem involvement with LCH. PMID- 11434844 TI - The epidemiology of pediatric traumatic brain injury in Minnesota. AB - OBJECTIVES: To determine the epidemiology of pediatric traumatic brain injury (TBI) in a midwestern state and to examine differences between metropolitan and nonmetropolitan residents. DESIGN: Population-based case series. PARTICIPANTS: Patients aged 0-19 years sustaining TBI in 1993 that resulted in hospitalization or death. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Incidence, mortality and case-fatality rates, length of hospital stay, discharge status, and Glasgow Outcome Scale score. RESULTS: Nine hundred seventy-seven patients met inclusion criteria. Incidence, mortality, and case-fatality rates were 73.5 per 100 000, 9.3 per 100 000, and 12.8 per 100, respectively. Higher median household incomes and percentages of adult high-school graduates in a patient's census block group correlated with lower incidence. Median length of stay was 2 days. Of those included in the study, 720 patients (74%) were discharged home with self-care. Three hundred fifty-seven patients met criteria for severe TBI; 346 (97%) were assigned Glasgow Outcome Scale scores, of which 161 (47%) had disabilities or died. Severe TBI was associated with nonmetropolitan residence, higher median household income, and certain injury mechanisms. Incidence was similar for metropolitan and nonmetropolitan residents. Median head-region Abbreviated Injury Score, Injury Severity Score, and mortality and case-fatality rates were higher for nonmetropolitan residents. CONCLUSIONS: This study reports the lowest incidence of pediatric TBI that results in death or hospitalization to date. One half of severely injured patients suffered poor outcomes. A greater proportion of nonmetropolitan than metropolitan residents suffered severe TBI and had higher mortality and case-fatality rates. PMID- 11434845 TI - Food-allergic reactions in schools and preschools. AB - BACKGROUND: Food allergies may affect up to 6% of school-aged children. OBJECTIVES: To conduct a telephone survey to characterize food-allergic reactions in children (defined as those aged 3-19 years in this study) with known food allergies in schools and preschools and to determine mechanisms that are in place to prevent and treat those reactions. DESIGN: The parents of food-allergic children were contacted by telephone and asked about their child's history of food-allergic reactions in school. The schools the children attended were contacted, and the person responsible for the treatment of allergic reactions completed a telephone survey. RESULTS: Of 132 children in the study, 58% reported food-allergic reactions in the past 2 years. Eighteen percent experienced 1 or more reactions in school. The offending food was identified in 34 of 41 reactions, milk being the causative food in 11 (32%); peanut in 10 (29%); egg in 6 (18%); tree nuts in 2 (6%); and soy, wheat, celery, mango, or garlic in 1 (3%) each. In 24 reactions (59%), symptoms were limited to the skin; wheezing occurred in 13 (32%), vomiting and/or diarrhea in 4 (10%), and hypotension in 1 (2%). Also, 15 (36%) of the 41 reactions involved 2 or more organ systems, and 6 (15%) were treated with epinephrine. Fourteen percent of the children did not have a physician's orders for treatment, and 16% did not have any medications available. Of the 80 participating schools, 31 (39%) reported at least 1 food-allergic reaction within the past 2 years and 54 (67%) made at least 1 accommodation for children with a food allergy, such as peanut-free tables, a peanut ban from the classroom, or alternative meals. CONCLUSIONS: It is common for food-allergic children to experience allergic reactions in schools and preschools, with 18% of children having had at least 1 school reaction within the past 2 years. Thirty six percent of the reactions involved 2 or more organ systems, and 32% involved wheezing. Every effort should be made to prevent, recognize, and appropriately treat food-allergic reactions in schools. PMID- 11434846 TI - Efficacy of naturopathic extracts in the management of ear pain associated with acute otitis media. AB - OBJECTIVE: To determine the efficacy and tolerance of Otikon Otic Solution (Healthy-On Ltd, Petach-Tikva, Israel), a naturopathic herbal extract (containing Allium sativum, Verbascum thapsus, Calendula flores, and Hypericum perforatum in olive oil), compared with Anaesthetic (Vitamed Pharmaceutical Ltd, Benyamina, Israel) ear drops (containing ametocaine and phenazone in glycerin) in the management of ear pain associated with acute otitis media (AOM). DESIGN: Children between the ages of 6 and 18 years who experienced ear pain (otalgia) and who were diagnosed with eardrum problems associated with AOM were randomly assigned to be treated with Otikon or Anaesthetic ear drops, which were instilled into the external canal(s) of the affected ear(s). Ear pain was assessed using 2 visual analog scales: a linear scale and a color scale. Pain assessment took place throughout the course of 3 days. The mean score of pain reduction was used to measure outcome. SETTING: Primary pediatric community ambulatory centers. PARTICIPANTS: One hundred three children aged 6 to 18 years who were diagnosed with otalgia associated with AOM. RESULTS: Each of the 2 treatment groups were comparable on the basis of age, sex, laterality of AOM, and the effectiveness of ameliorating symptoms of otalgia. The 2 groups were also comparable to each other in the initial ear pain score and in the scores at each application of Otikon or Anaesthetic drops. There was a statistically significant improvement in ear pain score throughout the course of the study period (P =.007). CONCLUSIONS: Otikon, an ear drop formulation of naturopathic origin, is as effective as Anaesthetic ear drops and was proven appropriate for the management of AOM-associated ear pain. PMID- 11434847 TI - Parent expectations for antibiotics, physician-parent communication, and satisfaction. AB - OBJECTIVES: To explore how parents communicate their preferences for antibiotics to their child's physician and to examine whether physicians can communicate why antibiotics are not being prescribed in a way that maintains satisfaction with the visit. DESIGN: Previsit survey of parents, audiotaping of the study encounters, and a postvisit survey of parents and physicians. SETTING: Two private pediatric practices. PARTICIPANTS: Ten physicians (response rate = 77%) and a consecutive sample of 295 eligible parents (response rate = 86%) who attended acute care visits for their children between October 1996 and March 1997. MAIN OUTCOME MEASURES: Physician-perceived pressure to prescribe antibiotics and parental visit-specific satisfaction. RESULTS: Fifty percent of parents expressed a previsit expectation for antibiotics. Among these parents, only 1% made a direct verbal request for them. Even when no direct requests for antibiotics were made, physicians still perceived an expectation for antibiotics 34% of the time. Among parents who did not receive expected antibiotics, those offered a contingency plan from the physician (i.e., the possibility of receiving antibiotics in the future if their child did not get better) had a higher mean satisfaction score than parents not receiving a contingency plan (76 vs. 58.9; P<.05). CONCLUSION: Physicians should consider providing a contingency plan to parents who expect antibiotics for their children when there is no clinical indication. Further study is needed to determine how parents indirectly communicate their desire for antibiotics and what additional communication techniques physicians can use to resist the overprescribing of antibiotics. PMID- 11434848 TI - The readability of pediatric patient education materials on the World Wide Web. AB - BACKGROUND: Literacy is a national and international problem. Studies have shown the readability of adult and pediatric patient education materials to be too high for average adults. Materials should be written at the 8th-grade level or lower. OBJECTIVE: To determine the general readability of pediatric patient education materials designed for adults on the World Wide Web (WWW). MATERIALS AND METHODS: GeneralPediatrics.com (http://www.generalpediatrics.com) is a digital library serving the medical information needs of pediatric health care providers, patients, and families. Documents from 100 different authoritative Web sites designed for laypersons were evaluated using a built-in computer software readability formula (Flesch Reading Ease and Flesch-Kincaid reading levels) and hand calculation methods (Fry Formula and SMOG methods). Analysis of variance and paired t tests determined significance. RESULTS: Eighty-nine documents constituted the final sample; they covered a wide spectrum of pediatric topics. The overall Flesch Reading Ease score was 57.0. The overall mean Fry Formula was 12.0 (12th grade, 0 months of schooling) and SMOG was 12.2. The overall Flesch Kincaid grade level was significantly lower (P<.0001), at a mean of 7.1, when compared with the other 2 methods. All author and institution groups had an average reading level above 10.6 by the Fry Formula and SMOG methods. CONCLUSIONS: Pediatric patient education materials on the WWW are not written at an appropriate reading level for the average adult. We propose that a practical reading level and how it was determined be included on all patient education materials on the WWW for general guidance in material selection. We discuss suggestions for improved readability of patient education materials. PMID- 11434849 TI - Adolescent cybersurfing for health information: a new resource that crosses barriers. AB - OBJECTIVE: To examine adolescents' use of and attitudes toward accessing health information through the Internet. DESIGN: Cross-sectional, school-based survey. PARTICIPANTS: A socioeconomically and ethnically diverse sample of 412 suburban New York 10th graders (mean [SD] age, 15.8 [0.68] years). MAIN OUTCOME MEASURES: Accessing the Internet for health information. RESULTS: Half (49%) of the sampled adolescents had used the Internet to get health information. Topics most often explored through the Internet included sexually transmitted diseases; diet, fitness, and exercise; and sexual behaviors. Adolescents found Internet information to be of high value (using a composite gauging worth, trustworthiness, use, and relevance), with no significant differences related to sex, ethnicity, or mother's education. When considering 11 separate health topics, girls found it more valuable to have information on birth control, diet and nutrition, exercise, physical abuse, sexual abuse, and dating violence. Only for alternative medicine were there differences by ethnicity, and there were no differences based on mother's education for the value of having specific health information available through the Internet. CONCLUSION: For adolescents, the Internet is an accessed and valued information source on a range of sensitive health issues. PMID- 11434850 TI - Do hair care practices affect the acquisition of tinea capitis? A case-control study. AB - OBJECTIVE: To determine the influences of hair-grooming practices and environmental factors as risk factors for the acquisition of tinea capitis (TC) in children. DESIGN: Case-control study comparing children with culture-proved TC with age-, sex-, and race-matched control subjects without scalp disease. SETTING: A multicenter study involving 3 urban referral centers in the United States. PARTICIPANTS: A convenience sample of 66 patients aged 12 years and younger presenting to pediatric dermatology clinics with clinical evidence of TC were enrolled as cases. Matched control subjects (n = 68), without known scalp disease, were enrolled from the outpatient pediatric clinics at the same institutions. RESULTS: Significant associations with TC in the conditional logistic regression model were a prior history of TC (odds ratio, 3.11; 95% confidence interval, 1.02-9.43; P =.04) and exposure to TC (odds ratio, 16.32; 95% confidence interval, 3.55-75.16; P =.001). The use of a hair conditioner was statistically significant in the univariable model but not in the multivariable model (odds ratio, 0.46; 95% confidence interval, 0.20-1.08; P =.07). Hairstyling, frequency of washing, use of oils or grease, and other hair care practices were not shown to be associated with the presence of TC. CONCLUSIONS: Hair-grooming practices do not appear to play a major role in the acquisition of TC. Hair conditioners may be protective in children at risk for TC, but further studies are needed to confirm this finding. PMID- 11434851 TI - The behavioral risks and life circumstances of adolescent mothers involved with older adult partners. AB - OBJECTIVE: To investigate behavioral risks and life circumstances of adolescent mothers with older (> or =5 years) adult (> or =20 years old) vs. similar-aged (+/-2 years) male partners at 12 months' postpartum. METHODS: Nine hundred thirty one adolescent females were interviewed after delivery and were mailed surveys to complete at 12 months' postpartum. Analysis by chi(2) and t test was used to identify differences in behavioral risks (planned repeated pregnancy, substance use, and intimate partner violence) and life circumstances (financial status, school enrollment, and social support) for adolescent mothers with older adult vs similar-aged partners. Additional stratified analyses were conducted to evaluate the extent to which living with an adult authority figure or being with the father of her infant born 12 months previously might alter observed relationships. RESULTS: At 12 months following delivery, 184 adolescent mothers (20%) reported having an older adult partner, whereas 312 (34%) had a similar aged partner. The remaining adolescent mothers (n = 239) were excluded from further analyses. Adolescent mothers with older adult partners were significantly less likely to be employed or enrolled in school and were more likely to report planned repeated pregnancies. These adolescent mothers also received less social support. No differences were observed in intimate partner violence or the mother's substance use. Adolescent mothers with older adult partners who did not live with an adult authority figure seemed to be at greatest risk. CONCLUSIONS: The negative educational and financial impact of coupling with an older vs. similar-aged partner seems greater for those mothers who no longer reside with an adult authority figure. These adolescent mothers are also at greater risk of planned rapid repeated pregnancy. Given their limited educational attainment and family support, a subsequent pregnancy may place these young women at considerable financial and educational disadvantage. PMID- 11434853 TI - Tobacco use outcomes of adolescents treated clinically for nicotine dependence. AB - OBJECTIVE: To evaluate the tobacco use outcomes and baseline characteristics of adolescents treated for nicotine dependence. DESIGN: Retrospective cohort study. SETTING: Mayo Clinic Nicotine Dependence Center, Rochester, Minn. PATIENTS: Ninety-six adolescents (60 boys, 36 girls) receiving clinical services for treatment of nicotine dependence between January 1, 1988, and November 30, 1997. Their mean age was 15.6 years (range, 11-17 years), and 91.7% were white. INTERVENTION: The Nicotine Dependence Center intervention involves a 45-minute consultation with a nicotine dependence counselor. A treatment plan individualized to the patient's needs is then developed. Telephone follow-up is conducted at 6 and 12 months. As part of this study, a long-term follow-up was conducted by telephone at a mean of 5.3 years (range, 1.6-10.6 years) following the intervention. MAIN OUTCOME MEASURES: Self-reported 7-day point-prevalence abstinence from tobacco at 6 and 12 months, and 30-day point-prevalence tobacco abstinence at the long-term follow-up. RESULTS: The tobacco abstinence rates were 17.7% (17/96 patients) at 6 months, 7.3% (7/96 patients) at 12 months, and 11.5% (11/96 patients) at the long-term follow-up. A high proportion of the sample had smoking-related medical morbidity and psychiatric diagnoses documented in the medical record prior to or at the time of the intervention. CONCLUSIONS: Adolescents utilize the medical community to seek treatment for nicotine dependence. The 6-month tobacco abstinence rate is higher than the estimates of the natural history of smoking cessation in adolescents. Medical and psychiatric diagnoses are common in this population. PMID- 11434854 TI - Evaluation of youth preferences for rapid and innovative human immunodeficiency virus antibody tests. AB - OBJECTIVE: To determine youth preferences for Food and Drug Administration (FDA) approved and investigational human immunodeficiency virus (HIV) antibody collection and testing methods before and after subjects learned of test result response times; to determine how influential test result response times are on participants' preferences. DESIGN: After health educators explained and demonstrated 6 different HIV antibody collection and testing strategies (3 saliva, 1 urine, and 2 fingerstick methods), participants completed a confidential survey about test method preference and tried the different testing methods. The participants had an opportunity to re-rank their test method preference after learning about each test's result response time. SETTING: Health education sessions in both clinical and community settings. PARTICIPANTS: Youths aged 12 to 24 years. RESULTS: An oral collection device with a rapid saliva test was the most highly preferred test method. The preference for this method and the rapid response test methods via fingerstick procedures improved significantly after subjects learned of the rapid result response time, while the other methods were given significantly lower preference rankings after subjects learned of the longer result response times. Shifts in preference rankings were not related to sex, age, ethnic group, experience with HIV testing, or practice of risk behaviors. CONCLUSIONS: Our research supports the use of noninvasive and rapid HIV testing methods with rapid response times for adolescents to assist in the early identification of HIV status, while offering HIV prevention opportunities and immediate linkage to care. PMID- 11434855 TI - Picture of the month. Multiple endocrine neoplasia 2B syndrome. PMID- 11434856 TI - Radiological case of the month. Cerebral air embolism in hereditary hemorrhagic telangiectasia. (Osler-Weber-Rendu disease). PMID- 11434857 TI - Radiological case of the month. Aneurysmal dilation of cerebral arteries associated with HIV infection. PMID- 11434858 TI - Pathological case of the month. Intracranial plasma cell granuloma. PMID- 11434859 TI - Pathological case of the month. Progressive hypertonic muscular dystrophy. PMID- 11434860 TI - Prevalence of adolescent varicocele. PMID- 11434861 TI - A successful strategy for increasing adherence to tuberculosis test reading in high-risk children. PMID- 11434862 TI - A needs assessment for establishing an after-hours telephone medicine curriculum. PMID- 11434863 TI - Pediatric counseling of parents who smoke. PMID- 11434865 TI - Counseling parents to quit smoking: little evidence of long-term success. PMID- 11434868 TI - Assessment of the usefulness of a diagnostic test: a survey of patient preference for diagnostic techniques in the evaluation of intestinal inflammation. AB - BACKGROUND: In order to assess the usefulness of radiolabeled white cell scanning in the diagnosis of intestinal inflammation, subjects were asked to rank several dimensions of preference for white cell scanning in relation to other diagnostic tests. Two groups were surveyed: one known to have inflammatory bowel disease and the second not familiar in most cases with the tests. Subjects were asked to rank preference for each of seven tests: radiolabeled white cell scan, colonoscopy, barium enema, sigmoidoscopy, enteroclysis, stool analysis and laparotomy for the diagnosis of IBD and impressions of discomfort, embarrassment, inconvenience and danger related to each test. Mean rank scores were calculated, test ranks compared within groups and significance determined by the Wilcoxon rank test. RESULTS: Significant differences were seen in overall preference for white cell scan over barium enema and colonoscopy (p < 0.01) in both survey groups. Perceived discomfort and embarrassment demonstrated similar rankings. CONCLUSION: This patient preference combined with the reported accuracy of white cell scanning further establishes the usefulness of this means of IBD diagnosis. PMID- 11434869 TI - Endothelial [Ca2+]i is an integrating signal for the vascular tone in rat aortae. AB - BACKGROUND: Although various endothelium-dependent relaxing factors (endothelial autacoids) are released upon the elevation of endothelial cytosolic free Ca2+ concentration (EC [Ca2+]i), the quantitative relationship between EC [Ca2+]i and vascular tone remains to be established. Moreover, whether the basal release of endothelial autacoids is modulated by basal EC [Ca2+]i is still unclear. We assessed these issues by using a novel method that allows simultaneous recording of EC [Ca2+]i and vascular displacement in dissected rat aortic segments. RESULTS: Receptor-dependent (acetylcholine) or independent (ionomycin) agonists caused immediate EC [Ca2+]i elevation followed by vasorelaxation in preparations pre-contracted with phenylephrine. Low doses of agonists induced small EC [Ca2+]i elevations (about 100 nmol/L) and concomitant half-maximal vasorelaxation. At high doses, agonists elevated EC [Ca2+]i to micromol/L range with little additional vasodilatation. When EC [Ca2+]i was plotted against the vasorelaxation, the curves were almost identical for both acetylcholine and ionomycin treatments, in the presence or absence of various endothelial autacoid inhibitors. Calcium-free solution reduced basal EC [Ca2+]i and induced a drastic vasoconstriction. Endothelial autacoid inhibitors reduced EC [Ca2+]i changes and abolished both agonist-induced vasodilatation and calcium-free solution-induced vessel contraction. When the EC [Ca2+]i was completely chelated by 40 micromol/L BAPTA, the acetylcholine-evoked vasorelaxation could be abolished as well. However, when the EC [Ca2+]i was partially chelated by 20 micromol/L BAPTA, the acetylcholine-evoked vasorelaxation was almost unaffected. CONCLUSIONS: These results indicate that vascular tone is modulated by subtle changes of EC [Ca2+]i level, which seems to serve as an integrating signal in both basal and stimulated states. PMID- 11434870 TI - Inducible transgenics. New lessons on events governing the induction and commitment in mammary tumorigenesis. AB - Breast cancer arises from multiple genetic events that together contribute to the established, irreversible malignant phenotype. The development of inducible tissue-specific transgenics has allowed a careful dissection of the events required for induction and subsequent maintenance of tumorigenesis. Mammary gland targeted expression of oncogenic Ras or c-Myc is sufficient for the induction of mammary gland tumorigenesis in the rodent, and when overexpressed together the rate of tumor onset is substantially enhanced. In an exciting recent finding, D'Cruz et al discovered tetracycline-regulated c-Myc overexpression in the mammary gland induced invasive mammary tumors that regressed upon withdrawal of c Myc expression. Almost one-half of the c-Myc-induced tumors harbored K-ras or N ras gene point mutations, correlating with tumor persistence on withdrawal of c Myc transgene expression. These findings suggest maintenance of tumorigenesis may involve a second mutation within the Ras pathway. PMID- 11434871 TI - The plasticity of human breast carcinoma cells is more than epithelial to mesenchymal conversion. AB - The human breast comprises three lineages: the luminal epithelial lineage, the myoepithelial lineage, and the mesenchymal lineage. It has been widely accepted that human breast neoplasia pertains only to the luminal epithelial lineage. In recent years, however, evidence has accumulated that neoplastic breast epithelial cells may be substantially more plastic in their differentiation repertoire than previously anticipated. Thus, along with an increasing availability of markers for the myoepithelial lineage, at least a partial differentiation towards this lineage is being revealed frequently. It has also become clear that conversions towards the mesenchymal lineage actually occur, referred to as epithelial to mesenchymal transitions. Indeed, some of the so-called myofibroblasts surrounding the tumor may have an epithelial origin rather than a mesenchymal origin. Because myoepithelial cells, epithelial to mesenchymal transition-derived cells, genuine stromal cells and myofibroblasts share common markers, we now need to define a more ambitious set of markers to distinguish these cell types in the microenvironment of the tumors. This is necessary because the different microenvironments may confer different clinical outcomes. The aim of this commentary is to describe some of the inherent complexities in defining cellular phenotypes in the microenvironment of breast cancer and to expand wherever possible on the implications for tumor suppression and progression. PMID- 11434872 TI - Tumour-stromal interactions. Role of the stroma in mammary development. AB - Mammary development depends on branching morphogenesis, namely the bifurcation and extension of ductal growth points (end buds) and secretory lobules into a more or less fatty stroma. Because breast carcinomas are overwhelmingly ductal in origin, this review focuses on stromal influences guiding postnatal ductal development and there is only the briefest account of the role of embryonic stroma (mesenchyme). The stroma as the necessary target for endocrine mammogens and the source of stimulatory growth factors is described and the importance of mammary epithelium-induced modifications of the periductal stroma is emphasized. Evidence is presented that if they are to grow, end buds must condition proximal fatty stroma by recruiting white blood cells as well as inducing stromal cell division and, possibly, estrogen receptors. The induction of a fibrous stromal tunic around the end bud is described and its likely role as a complex ductal morphogen is discussed; a possible role in growth inhibition is also considered. Although the signals governing fibrotic induction, ductal morphogenesis, and growth inhibition are unknown, a role for transforming growth factor-beta is highly likely and is discussed. Finally, a need for new conceptual and experimental approaches to understanding stromal-epithelial signaling is discussed. PMID- 11434873 TI - Tumour-stromal interactions. Integrins and cell adhesions as modulators of mammary cell survival and transformation. AB - Stromal-epithelial interactions modulate mammary epithelial cell (MEC) growth and apoptosis by influencing cell adhesion and tissue organization. Perturbations in the mammary stroma and cell adhesion characterize breast tumors and underlie the altered tissue organization, disrupted tissue homeostasis and enhanced survival phenotype of the disease. Apoptosis resistance likely arises during malignant transformation via genetic and epigenetic modification of cell adhesion pathways induced by a changing tissue microenvironment. Acquisition of adhesion-linked survival networks that enhance MEC viability in the absence of basement membrane interactions probably promote malignant transformation, and may render breast tumors sufficiently resistant to exogenous apoptotic stimuli to generate multidrug resistance. PMID- 11434874 TI - Tumour-stromal interactions. Transforming growth factor-beta isoforms and hepatocyte growth factor/scatter factor in mammary gland ductal morphogenesis. AB - The mammary gland undergoes morphogenesis through the entire reproductive life of mammals. In mice, ductal outgrowth from the nipple across the fat pad results in an intricate, well spaced ductal tree that further ramifies and develops alveolar structures during pregnancy. Ductal morphogenesis is regulated by the concerted action of circulating steroid and polypeptide hormones, and local epithelial mesenchymal inductive signals. Transforming growth factor (TGF)-beta1-3 and hepatocyte growth factor (HGF)/scatter factor (SF) are important components of this latter signaling pathway. TGF-beta1 and TGF-beta3 have roles in both promotion and inhibition of branching morphogenesis that are dependent on concentration and context. HGF/SF promotes ductal outgrowth and tubule formation in the mammary gland. These data suggest that these two growth factors have complementary roles in promoting mammary ductal morphogenesis and in maintaining ductal spacing. In addition, TGF-beta3 triggers apoptosis in the alveolar epithelia, which is a necessary component of mammary gland involution and return of the ductal structure to a virgin-like state after lactation. PMID- 11434875 TI - Prostate-specific antigen (PSA/hK3): a further player in the field of breast cancer diagnostics? AB - Since its identification, much information has been obtained about prostate specific antigen (PSA, or human glandular kallikrein 3 [hK3]), a kallikrein-like serine protease that is the most valuable tumour marker for the screening, diagnosis and management of human prostate carcinoma. Recently, it has become widely accepted that PSA is also present in many nonprostatic sources, casting doubts about the specificity of its tissue expression. Here we summarize the findings on the biomolecular expression of PSA in breast secretions, cells and tissues of healthy and diseased females. Although several studies have strongly suggested that the molecular forms of PSA seem to represent a potential tool for the risk assessment of breast cancer, recent reports have yielded conflicting results. Although several studies have suggested new biological function(s) for PSA in breast physiopathology, more studies are needed to enlist PSA unequivocally as an additional weapon in the anticancer armoury in breast cancer diagnostics. PMID- 11434876 TI - Reversal effects of nomegestrol acetate on multidrug resistance in adriamycin resistant MCF7 breast cancer cell line. AB - BACKGROUND: Chemotherapy is important in the systematic treatment of breast cancer. To enhance the response of tumours to chemotherapy, attention has been focused on agents to reverse multidrug resistance (MDR) and on the sensitivity of tumour cells to chemical drugs. Hundreds of reversal drugs have been found in vitro, but their clinical application has been limited because of their toxicity. The reversal activity of progestogen compounds has been demonstrated. However, classical agents such as progesterone and megestrol (MG) also have high toxicity. Nomegestrol (NOM) belongs to a new derivation of progestogens and shows very low toxicity. We studied the reversal activity of NOM and compared it with that of verapamil (VRP), droloxifene (DRO), tamoxifen (TAM) and MG, and investigated the reversal mechanism, i.e. effects on the expression of the MDR1, glutathione S transferase Pi (GSTpi), MDR-related protein (MRP) and topoisomerase IIalpha (TopoIIalpha) genes, as well as the intracellular drug concentration and the cell cycle. The aim of the study was to examine the reversal effects of NOM on MDR in MCF7/ADR, an MCF7 breast cancer cell line resistant to adriamycin (ADR), and its mechanism of action. METHODS: MCF7/ADR cells and MCF7/WT, an MCF7 breast cancer cell line sensitive to ADR, were treated with NOM as the acetate ester. With an assay based on a tetrazolium dye [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H tetrazolium bromide; MTT], the effects of various concentrations of NOM on MDR in MCF7/ADR cells were studied. Before and after the treatment with 5 microM NOM, the expression of the MDR-related genes MDR1, GSTpi, TopoIIalpha and MRP were assayed with a reverse transcriptase polymerase chain reaction (RT-PCR) immunocytochemistry assay. By using flow cytometry (FCM), we observed the intracellular ADR concentration and the effects of combined treatment with NOM and ADR on the cell cycle. Results collected were analysed with Student's t test. RESULTS: NOM significantly reversed MDR in MCF7/ADR cells. After treatment NOM at 20, 10 and 5 microM, chemosensitivity to ADR increased 21-fold, 12-fold and 8 fold, respectively. The reversal activity of NOM was stronger than that of the precursor compound MG, and comparable to that of VRP. After treatment with 5 microM NOM, the expression of both the MDR1 and the GSTpi mRNA genes began to decline on the second day (P <0.05 and P <0.01, respectively), and reached the lowest level on the third day (both P <0.01); however, on the fifth day the expression levels began to increase again (both P <0.05). The expression of MRP and TopoIIalpha had no significant changes. Changes in the expression of P glycoprotein (P-gp) and GSTpi were similar to those of their mRNA expressions, showing early declines and late increases. Two hours after treatment with 20, 10 and 5 microM NOM, the intracellular ADR concentration increased 2.7-fold, 2.3 fold and 1.5-fold respectively. However, NOM did not increase ADR accumulation in MCF7/WT cells. FCM data showed that after 48 h of combined administration of NOM (20 microM) and ADR (from low to high concentration), MCF7/ADR cells showed a gradual arrest at the G2M phase with increasing ADR dose. The arrest effect with combined drug treatment was stronger than that with the single ADR treatment. CONCLUSION: MDR is the major mechanism of drug resistance in malignant tumour cells. To overcome MDR and to increase chemosensitivity, many reversal agents have been found. Most progestogen compounds have been demonstrated to have reversal effects, but we found no data on NOM, a new progestogen compound. Our results show that NOM has strong reversal activity. The reversal effects were stronger than those of the precursor compound, MG, and were comparable to that of VRP. Because NOM has low toxicity, it might have good prospects in clinical application. Using RT-PCR and immunocytochemistry assays, we studied the effects of NOM on MDR-related genes. The results were that NOM could markedly downregulate the mRNA and protein expression levels of MDR1 and GSTpi. TopoIIalpha and MRP gene expression showed no significant changes. It is known that P-gp induces MDR in tumour cells mainly by decreasing the intracellular drug concentration. After treatment with NOM, the intracellular drug concentration in MCF7/ADR cells increased significantly. Combined treatment with NOM and ADR induced arrest at the G2M phase. It is worth noting that NOM caused an early decrease and a late increase in the expression of some MDR-related genes in a time-dependent manner. The phenomena raise a question for the continued administration of reversal agents in clinics that merits further study. We demonstrate that NOM has strong reversal effects on MDR in MCF7/ADR cells. The reversal is via different routes, namely downregulating the mRNA and protein expression levels of MDR1 and GSTpi, increasing intracellular drug concentration and arresting cells at the G2M phase (NOM in combination with ADR). The reversal mechanism needs further study. PMID- 11434877 TI - Possible association of beta2- and beta3-adrenergic receptor gene polymorphisms with susceptibility to breast cancer. AB - BACKGROUND: The involvement of beta2-adrenergic receptor (ADRB2) and beta3 adrenergic receptor (ADRB3) in both adipocyte lipolysis and thermogenic activity suggests that polymorphisms in the encoding genes might be linked with interindividual variation in obesity, an important risk factor for postmenopausal breast cancer. In order to examine the hypothesis that genetic variations in ADRB2 and ADRB3 represent interindividual susceptibility factors for obesity and breast cancer, we conducted a hospital-based, case-control study in the Aichi Cancer Center, Japan. METHODS: A self-administered questionnaire was given to 200 breast cancer patients and 182 control individuals, and pertinent information on lifestyle, family history and reproduction was collected. ADRB2 and ADRB3 genotypes were determined by polymerase chain reaction (PCR) restriction fragment length polymorphism assessment. RESULTS: Twenty-five (12.4%) breast cancer patients and 32 (17.6%) control individuals were found to bear a glutamic acid (Glu) allele for the ADRB2 gene (odds ratio [OR] 0.67, 95% confidence interval [CI] 0.38-1.18), and 60 (30.0%) breast cancer patients and 61 (33.5%) control individuals were found to bear an Arg allele for the ADRB3 gene (OR 0.85, 95% CI 0.55-1.31). A significantly lower risk was observed in those who carried the Glu ADRB2 allele and who reported first childbirth when they were younger than 25 years (OR 0.35; 95% CI 0.13-0.99). CONCLUSION: A potential association may exist between risk of breast cancer and polymorphisms in the ADRB2 and ADRB3 genes; further studies in larger samples and/or in different ethnic groups are warranted to investigate this potential association. PMID- 11434878 TI - Breast cancer mortality among Ashkenazi Jewish women in Sao Paulo and Porto Alegre, Brazil. AB - BACKGROUND: Increased BRCA1 and BRCA2 germline mutation rates have been reported in Ashkenazi Jewish women in North America, Europe and Israel, and have been mentioned as possibly related to a higher incidence of breast and ovarian cancer among these communities. The present study was carried out with the aim of obtaining evidence on the magnitude of breast cancer as a cause of death among Ashkenazi women in Brazil. METHODS: We reviewed all death certificates archived in the Jewish Burial Societies of Sao Paulo (1971-1997) and Porto Alegre (1948 1997), two of the main and oldest Jewish communities in Brazil. Breast cancer observed deaths were compared with expected deaths according to breast cancer mortality in the general population. RESULTS: The observed ratios were approximately quite close to unity, suggesting a similar breast cancer mortality pattern among the Ashkenazi population and the general population in both cities. These results maintain similar behavior regardless of whether analyzed before or after the mid-1980s, when mammography came to be increasingly performed in Brazil. Cancer proportional mortality ratios were 1.04 (0.83-1.29) in Sao Paulo and 1.16 (0.84-1.57) in Porto Alegre before 1985, and 1.17 (1.00-1.44) and 1.21 (0.81-1.79), respectively, between 1985 and 1997. Some evidence of the maintenance of protective risk factors such as high parity has been observed among Ashkenazi women in Sao Paulo. CONCLUSION: A quite similar breast cancer mortality pattern was observed between Ashkenazi Jewish women and the general population in Sao Paulo and Porto Alegre, Brazil. These results may suggest an environmental role on germ mutation expression reported in this ethnic group. PMID- 11434879 TI - In vivo cell kinetics in breast carcinogenesis. AB - BACKGROUND: Disruption of the balance between apoptosis and proliferation is considered to be an important factor in the development and progression of tumours. In the present study we determined the in vivo cell kinetics along the spectrum of apparently normal epithelium, hyperplasia, preinvasive lesions and invasive carcinoma, in breast tissues affected by fibrocystic changes in which preinvasive and/or invasive lesions developed, as a model of breast carcinogenesis. MATERIALS AND METHODS: A total of 32 areas of apparently normal epithelium and 135 ductal proliferative and neoplastic lesions were studied. More than one epithelial lesion per case were analyzed. The apoptotic index (AI) and the proliferative index (PI) were expressed as the percentage of TdT-mediated dUTP-nick end-labelling (TUNEL) and Ki-67-positive cells, respectively. The PI/AI (P/A index) was calculated for each case. RESULTS: The AIs and PIs were significantly higher in hyperplasia than in apparently normal epithelium (P = 0.04 and P = 0.0005, respectively), in atypical hyperplasia than in hyperplasia (P = 0.01 and P = 0.04, respectively) and in invasive carcinoma than in in situ carcinoma (P < 0.001 and P < 0.001, respectively). The two indices were similar in atypical hyperplasia and in in situ carcinoma. The P/A index increased significantly from normal epithelium to hyperplasia (P = 0.01) and from preinvasive lesions to invasive carcinoma (P = 0.04) whereas it was decreased (non-significantly) from hyperplasia to preinvasive lesions. A strong positive correlation between the AIs and the PIs was found (r = 0.83, P < 0.001). CONCLUSION: These findings suggest accelerating cell turnover along the continuum of breast carcinogenesis. Atypical hyperplasias and in situ carcinomas might be kinetically similar lesions. In the transition from normal epithelium to hyperplasia and from preinvasive lesions to invasive carcinoma the net growth of epithelial cells results from a growth imbalance in favour of proliferation. In the transition from hyperplasia to preinvasive lesions there is an imbalance in favour of apoptosis. PMID- 11434881 TI - ? PMID- 11434880 TI - Silver anniversary of the Spanish Society of Clinical Immunology and Pediatric Allergology. PMID- 11434882 TI - [Primary immunodeficiencies. Clinical features and variant forms]. AB - Periodically the World Health Organization and currently the International Union of Immunology Societies publish a classification of primary immunodeficiency diseases (PID) that includes diagnostic and therapeutic guidelines. The latest of these publications dates from 1999 and includes a new group of PID, the proliferative autoimmune syndromes. Furthermore, new forms of severe combined immunodeficiency (SCID) and of recessive autosomal agammaglobulinemia are described. From the publication of this classification until the end of the year 2000 a minimum of three new PIDs have been described and a further two should probably be added. Progress in the molecular biology of these diseases has given rise not only to more accurate diagnosis but also to greater insight into the clinical spectrum of these diseases. A mutation or deletion in a gene can provoke the complete absence of its product; sometimes expression is partial or normal but functional activity is absent or defective. In certain cases, partial or defective activity causes variant forms of the disease presenting symptomatology or atypical cellular phenotype. In other cases, this is not cause of the variant form, which can appear in interfamilial cases sharing the same mutation. In these cases, these differences can be attributed to environmental factors or to other genes able to modify the affected gene. In this article we provide examples of variant forms in several PIDs. Some are late onset forms, such as X-linked agammaglobulinemias diagnosed in adults, since until diagnosis, clinical symptomatology was minimal. In adenosine-deaminase deficiency, a serious and highly lymphoproliferative form of SCID, patients have been described whose symptomatology began after the age of 20 years. Another SCID, RAG1 and RAG2 recombinase deficiency, may produce a typical form with a characteristic T-B-NK + phenotype, Omenn's syndrome, or forms with an unexpected T-B + NK + phenotype. Deficiency in common gamma chain receptor for IL-2 may produce phenotypical variants that can lead to diagnostic error. X-linked lymphoproliferative syndrome may present as fulminant infectious mononucleosis, as leukemia or lymphoma or as hipo- or agammaglobulinemia. Possibly, some patients diagnosed with common variable immunodeficiency or with x-linked agammaglobulinemia do in fact have this syndrome. Chronic granulomatous disease is usually of early-onset, but late onset forms have been described. In one case the first clinical manifestation was produced when the patient was 60 years old. The above examples serve to highlight that, even though PIDs are usually suspected by pediatricians, in some cases the diagnosis may be missed by internists or non-pediatricians. Moreover, the clinical and laboratory findings of these variant forms must be determined to carry out an early diagnosis, which is essential for a favorable therapeutic outcome. PMID- 11434883 TI - [Molecular diagnosis of primary immunodeficiencies]. AB - Knowledge of the molecular defects responsible for some primary immunodeficiency diseases (PIDs) offers undoubted advantages in establishing a reliable diagnosis. Such knowledge would allow us not only to establish a prognosis but also to instigate the most appropriate therapy. After molecular diagnosis, some patients could benefit from gene therapy. However, apart from the diagnosis of the disease, molecular biological techniques also enable more reliable identification of carriers and, when suggested by the family history and when the familial defect is already known, prenatal diagnosis will also be possible, thus establishing the earliest possible treatment. Using the single-stranded conformational polymorphism technique followed by direct sequencing, we found 22 different mutations in 22 patients from unrelated families and with a phenotype compatible with x-linked agammaglobulinemia. Fourteen of these are new, previously undescribed mutations and the remaining eight are already included in the data base (http://www.uta.fi/imt/bioinfo/Btkbase). Analysis of the female carrier was performed in all the mothers and the mutation was de novo in only one patient. Study of the BtK gene enabled differential diagnosis with common variable immunodeficiency disease in some patients who showed absent or very low lymphocyte B counts as well as forms of autosomal recessive agammaglobulinemia. Using the same techniques, we were able to identify mutations in the CD40 ligand gene in three families in which one of the members had clinical and biological phenotype compatible with X-linked hyper-IgM. Molecular diagnosis was very useful in identifying carriers in these families as well as in making the differential diagnosis among patients with common variable immunodeficiency disease. Purely on this were we able to provide appropriate genetic counseling. PMID- 11434884 TI - [Common variable immunodeficiency. Review]. AB - Common variable immunodeficiency (CVI) is a primary immunodeficiency characterized by deficient antibody production. The cause of this immunodeficiency is unknown; several in vitro studies have revealed a significant number of alterations that could explain the hypogammaglobulinemia present in this syndrome. Among those described are primary B cell alterations, numerical and functional T cell abnormalities, and defects in the interaction between accessory cells. The alteration typical of CVI is the failure of B lymphocytes to differentiate from antibody-producing cells, resulting in deficient immunoglobulin secretion. Among the T cell abnormalities described are a diminished proliferative response to mitogens and antigens, alterations in the level of production of several cytokines, especially reduction in the production of IL-2, diminished antigen-specific T cells and increase basal apoptosis after stimulation. Antigen presenting cells, monocytes and dendritic cells can also present alterations and contribute to deficient antigen response. The clinical manifestations of these patients is variable; most present recurrent bacterial infections due to encapsulated bacteria, especially sinusitis, otitis, bronchitis, and pneumonias. A few patients can present mycobacterial or fungal infection and occasionally Pneumocystis carinii. Viral infection is uncommon in these patients although some suffer recurrent herpes zoster infection. Clinical features of septicemia and central nervous system infections are less frequent. The incidence of digestive tract infections in these patients is high. The most common cause of diarrhea is Giardia lamblia; Salmonella, Shigella and Campylobacter are also common pathogens. Autoimmune disease is also more prevalent in these patients than in the general population. The most frequently associated diseases are hemolytic anemia, idiopathic thrombocytopenic purpura and autoimmune neutropenia. Cancer is also frequently associated with CVI, the most common forms being lymphoproliferative syndromes, especially non-Hodgkin's lymphoma. Granulomas are a unusual manifestation in some patients with CVI; their localization varies but the most commonly affected organs are the spleen and lungs. Some authors have compared these granulomas with those characterizing sarcoidosis, especially when appearing in the lung. Diagnosis of CVI is usually by exclusion of other diseases, such as cystic fibrosis, immotile cilia syndrome or allergic processes. CVI should be suspected in all patients with recurrent bacterial infections especially those localized in the respiratory tract. Other primary immunodeficiencies which present clinical findings similar to CVI and which should be ruled out are selective IgG subclass deficiency, IgA deficiency and selective deficiency in the response to polysaccharide antigens with normal immunoglobulin levels. The serum hypogammaglobulinemia present in all patients with CVI provides the diagnostic key. The age at which clinical manifestations appear, the absence of familial antecedents and the presence of circulating B lymphocytes form the basis of the differential diagnosis between X-linked agammaglobulinemia and autosomal recessive forms. The treatment of choice of patients with CVI is treatment with human gamma-globulin. Currently, the most common route of administration is intravenous; these molecules have a half-life of approximately 21 days and a high degree of safety concerning the possible transmission of viral infections. Adverse reactions are generally few and clinically unimportant. The most frequently used doses oscillate between 200 and 400 mg/kg body weight every 2-4 weeks. Both the dose and its frequency should be personalized for each patient. Early diagnosis of patients with CVI, application of treatment with appropriate antibiotics for infections and treatment with gamma globulins prevent long-term complications of this disease and dramatically improve the quality of life and life expectancy of these patients. PMID- 11434885 TI - [Substitution therapy with hematopoietic progenitors in the primary immunodeficiencies]. AB - Hematopoietic stem-cell transplantation is currently the most appropriate substitution therapy in the most severe forms of primary immunodeficiency diseases (all the variants of SCID, WA, CID etc.). It can achieve total and permanent immunological reconstitution in 60% of patients, depending on histocompatibility, source of the hematopoietic stem cells and the underlying disease. Stem-cell sources may be bone marrow, umbilical cord blood and the peripheral blood of donors previously treated with colony stimulating factors for the mobilization CD34. We discuss the differences in the results obtained in patients treated at the Hospital Materno-Infantil Vall d'Hebron. Gene therapy opens a new era in the treatment of primary immunodeficiency diseases. The first patient to undergo this treatment in the United States of America had adenosine deaminase deficiency, even though sustained remodeling has not been achieved. The favorable results obtained in patients with SCID by deficit in the gamma chain of the IL-2 receptor in Paris, with more than a year of follow up, suggest that the near future is promising. We also discuss the differences observed according to the vectors used and the underlying disease. PMID- 11434886 TI - [The Spanish Registry of Primary Immunodeficiencies (REDIP)]. AB - Two thousand and fifty cases (n = 2050) of primary immunodeficiencies (PID) were registered up to February 2001. The Spanish Register for Primary Immunodeficiencies (REDIP) began in 1993. PID nomenclature and diagnostic criteria were made according to the report of the World Health Organization Scientific Group (1999). The most frequent disorders were IgA deficiency (797 registers) and common variable immunodeficiency (CVI) (389), followed by severe combined immunodeficiency and predominantly T cell defects (268), complement deficiencies (207 registers), X-linked agammaglobulinemia (87), IgG subclass deficiency (71), chronic granulomatous disease (64). Gammaglobulin replacement was the therapy in 638 patients (76%) belonging to antibody deficient group. 61 bone marrow transplants were done, 46 severe combined immunodeficiencies, 6 phagocytic disorders and 1 unclassified. Important differences in the number of cases submitted from different country areas were found. PMID- 11434887 TI - [Viral infection and asthma: immunologic mechanisms]. AB - The role of viral respiratory infections in lactating infants and other children continues to generate controversy. The debate concerns the difference, or the apparent differences, in the natural history of wheezing. Viral infections frequently provoke wheezing episodes in non-asthmatic small children but in the majority of these the wheezing disappears without the child subsequently developing asthma. In some cases, however, the wheezing persists and in others the child has asthma. Both the role of viral infection and the mechanisms by which wheezing can be produced in a previously healthy child or exacerbated in asthmatic children are unknown. Several hypotheses have been put forward to explain the relationship between viral infections and persistent wheezing and asthma: 1. Altered immune response to various allergens, whether producing sensitization to these allergens or inhibiting tolerance response to airborne allergens. The number of such patients is increasing, among them those with bronchiolitis, asthma, positive skin tests and specific IgE antibodies. Although there is no unanimity on the matter, these patients also present elevated IL-4 levels and reduced IFN-gamma levels. 2. Induction of inflammation typical of allergic asthma. This occurs when the virus interacts with T lymphocytes; (the natural response to viral infection is Th0 and Th1 lymphocyte differentiation and release of IFN-gamma, which has antiviral properties. In children infected with respiratory syncytial virus Th2 lymphocyte differentiation is produced, which is characteristic of allergic reactions, to the detriment of Th1); epithelial cells (in these cells active viral infection activates nuclear transcription kappa-beta and nuclear IL-6 factor, producing the release of numerous pro-inflammatory cytokines and chemokines as well as expression of adhesion molecules); eosinophils (inducing variable eosinophilia which, to a certain degree, has predictive value for the persistence of wheezing) and other inflammatory cells such as neutrophils and macrophages. In the same context, during viral respiratory infection, the presence of mediators (leukotrienes, especially LTC4, histamine, prostaglandins and tryptase) are observed in respiratory secretions and a correlation between levels of specific IgE mediators can be observed. 3. Increased allergic inflammation--producing bronchial hyperreactivity, mediator release by the various inflammatory cells and neuropeptides from C-sensitive fibers, and even interfering with nitric oxide bronchodilators. In spite of all of the above, it seems that recurrent wheezing after childhood bronchiolitis is not exclusively the result of viral infection and that other factors also play a role in this disease. PMID- 11434888 TI - [Childhood asthma and viral infection: interactions and therapeutic possibilities]. AB - Rhinovirus infections are the main cause of wheezing in children and adults. Studies carried out with experimental infections report that at least in certain conditions, this infection may spread to the lower respiratory tract as the virus acts on the respiratory epithelium. In vitro experiments with cells from the immune system and lower respiratory tract suggest that the mechanisms of action are directly linked to the production of pro-inflammatory cytokines. Both in vivo and in vitro evidence shows that rhinoviruses may stimulate bronchial epithelial cells to produce cytokines and pro-inflammatory chemokines. They may also stimulate the cholinergic and non-cholinergic nervous system, increasing the production of ICAM-1 and may give rise to a T-lymphocyte non-specific response or to T-lymphocyte replication in direct relation with viral infection. In addition, greater production of cysteinyl leukotrienes has been observed in the secretions of patients with bronchospasm. Experimental infection with Rhinovirus increases clinical symptomatology and bronchial hyperreactivity. The latter is associated with increased eosinophils and cationic proteins in sputum. The above findings suggest that multiple cellular pathways are involved in the induction of exacerbations of asthma induced by the virus. In addition, the possible stimulating role of exposure to allergens in sensitized patients would also increase allergic inflammation. Because of the limited therapeutic efficacy of steroids in reducing exacerbations, new treatment strategies based on greater insight into the physiopathology of the role of viruses in asthma are needed. PMID- 11434889 TI - [Respiratory syncytial virus infection and asthma]. AB - Presentation of bronchial asthma, in the years following the first outbreak of bronchiolitis due to respiratory syncytial virus (RSV) was first described by McIntosh, who postulated a common pathogenesis that was confirmed by the greater frequency (50%) of wheezing bronchiolitis and asthma during the more than 5 year follow up of these children. More recently, Hibbert and Schroechkenstein have again confirmed this phenomenon. These authors report that the percentage of asthma was increased by up to 71% in a group of children who contracted bronchiolitis during the first year of life and who were closely followed-up for 5 years after the outbreak. Other authors report figures between 25% and 57%. Stein et al. followed-up 888 children with RSV bronchiolitis until the age of 13 years and observed that at the age of 3-5 years 69% had asthma, at 4-5 years 55% did so and at 6-8 years 31% were asthmatic. In our experience of children who developed RSV bronchiolitis before the age of 6 months, 58 of 75 developed infantile asthma in following 3 years. Seventeen infants were aged more than 6 months at onset of bronchiolitis and of these 5 had bronchiolitis. We carried out a prospective study of 50 children aged 3-7 months with RSV bronchiolitis from December 1997 to February 1998. Follow-up was until the year 2000. Of these children, 22 (44%) had asthma and the remaining 28 (56%) had isolated episodes of cough and wheezing, which did not fulfill the criteria for asthma. Of the 22 children with asthma, all presented elevated total IgE by the second year of follow-up but only one of the children presented hypersensitivity to egg. The breathing difficulties that appeared in the initial outbreak of bronchiolitis is well explained by the cytopathic effect of the virus on the airways of infants. RSV virus produces inflammation of the bronchial mucosa, the effects of which may persist for 6-7 weeks, even after recovery from the first episode. The damaged and denuded epithelium provides fertile ground for future viral reinfections which, although less severe, produce dyspnea due to irritation of the exposed vagal receptors. Irrespective of the mechanism involved (RSV, histamine, methacholine or ozone inhalation) inflammation alters the intercellular junctions of the bronchial mucosal epithelium which, even in the absence of significant necrotic lesions, leads to increased penetration of irritants and stimulates the vagal receptors, leading to bronchospasm. In conclusion, it can currently be stated that RSV bronchiolitis constitutes a risk factor for the development of infantile asthma. The risk is increased in children with familial or personal antecedents of atopy. Serious forms of bronchiolitis due to RSV are more frequent in atopic individuals. The appearance of extrinsic asthma is more frequent in children who have previously had RSV bronchiolitis than in those who have not. PMID- 11434890 TI - [The role of infection in asthma]. AB - Since the first decades of the twentieth century, some authors have believed bacterial respiratory infection to be an important triggering factor in bronchial asthma, drawing attention to an asthmatic response to infection. In this context, already in 1995, we presented a study on nasal secretion cultures and the relationship between IgE and sensitization to allergens. There was a statistically significant association between patients with sensitization to Dermatophagoides, elevated IgE levels and Staphylococcus Aureus positive cultures. Following the studies by Norn, we performed a study in 40 children, aged 2-14 years, and observed that these children with sensitization to mites and positive culture released higher histamine levels than did children with negative cultures and controls. The differences were statistically significant. In agreement with other authors, we also found that the presence of both S. aureus and D. pteronyssinus favored antigen specific histamine release. In the last few years, when the increase in the prevalence of bronchial asthma began to be studied, the role of infection, among other factors, in favoring this increase began to be examined. Using the methodology of the ISAAC project, we distributed a parallel questionnaire containing questions on triggering and contributing factors among which was respiratory infection. We found that there was an association between having three of more episodes of bronchitis in the previous year, accompanied by fever and with a duration of more than 7 days and having asthma at some time (OR: 29.09). This association was even higher in patients with wheezing in the previous 12 months (OR: 43.26) and was also associated with the need to present to the emergency department (OR: 30.65). From these results we conclude that respiratory infection is an aggravating factor in asthma, as we already know. For several years, several authors have studied how non-nosocomial respiratory infections can directly modulate Th1/Th2 response. In order to obtain our own results, we studied serum interleukin 4 (IL4) and interferon-gamma (IFN gamma) in 42 children aged 3-17 years. The most frequent IL-4 values expressed in ng/ml were between 0.25-0.40, with little variation in the sample, which did not permit correlation among variables. Concerning IFN-gamma, we found values between < 5 and 605 pg/ml. In children undergoing antigen-specific immunotherapy, we observed mean IFN-gamma values of 115.86 pg/ml, while children not undergoing immunotherapy and those who had been administered this treatment for less than 1 year, had a mean of 66.06 pg/ml. These differences were statistically significant (p = 0.035), thus revealing a Th1 response to immunotherapy. These differences were not statistically significant when children who had been administered immunotherapy for less than 1 year were included. When we studied children with bacterial immunotherapy and grouped them in the same way, we found that the mean IFN-gamma of the children undergoing immunotherapy for more than 1 year was 56.4 pg/ml compared with 101.75 pg/ml in those without immunotherapy. This difference was statistically significant (p = 0.034). We are able to conclude that bacterial immunotherapy modifies Th1 response, inhibiting it in children with higher susceptibility to infection. In view of these preliminary results, it would be interesting to continue to study interleukins in order to determine the modification of these substances by immunotherapy in a prospective study and with a sample selected in relation to immunotherapy and not other parameters, since those we have studied have shown no relationship. PMID- 11434892 TI - ? PMID- 11434891 TI - [The past, present, and future of the Spanish Society for Clinical Immunology and Pediatric Allergology]. PMID- 11434894 TI - Intermittent claudication: magnitude of the problem, patient evaluation, and therapeutic strategies. AB - Intermittent claudication (IC), the symptom of exercise-induced muscle ischemia of peripheral arterial disease (PAD), afflicts and limits the activities of a significant number of patients. Incidence and prevalence of IC depends on the population studied and the diagnostic instruments used. In large studies, prevalence has ranged from 3% to 10%, with a sharp increase in those aged > or =70 years. Over the next 20 years, the total number of patients affected is expected to increase significantly due to anticipated demographic changes. Analysis of the natural history of IC demonstrates that the risk of cardiovascular morbidity and mortality far exceeds that of severe limb ischemia or limb loss. In fact, only 2% to 4% of all patients with IC will require a major amputation in their lifetime. However, life expectancy is approximately 10 years less than that of an age-matched cohort. By now, PAD is well recognized as a marker of systemic atherosclerosis. The cornerstone of patient evaluation is a history and physical examination, including a detailed atherosclerotic risk factor assessment. In the differential diagnosis of IC, clinicians should consider etiologies such as arthritis, spinal stenosis, radiculopathy, venous claudication, or inflammatory processes. In >80% of all patients, it is possible to locate the responsible arterial segment by combining the location and severity of pain with a pulse examination. Noninvasive diagnostic studies help determine the level of disease, may unmask a hemodynamically significant stenosis, and are useful in follow-up. Arteriography is reserved for patients in whom the decision for revascularization has been made. Knowing the anatomic detail of a lesion allows the clinician to determine whether and what type of intervention is feasible. Standard therapy for all patients should be directed at both peripheral and systemic atherosclerosis, beginning with risk-factor modification in the form of smoking cessation, optimal diabetes control, and lipid normalization. The benefits of supervised exercise rehabilitation include significantly increased walking distance and enhanced quality of life. Platelet inhibition has been shown to reduce the risk of ischemic stroke, myocardial infarction, and vascular death and should be prescribed for all but those in whom it is medically contraindicated. Symptom-specific pharmacotherapy with a broad range of medications has yielded disappointing results in the past. However, recent studies have demonstrated that patients receiving the novel agent cilostazol experienced increases in walking distance and improvements in quality of life. PMID- 11434895 TI - Intermittent claudication: effective medical management of a common circulatory problem. AB - Intermittent claudication (IC), most often characterized by a reproducible, painful aching or cramping in muscle groups of the leg caused by walking and relieved by rest, is a common, lifestyle-limiting symptom of lower-extremity peripheral arterial occlusive disease. Because IC is usually indicative of systemic atherosclerosis, active investigation and treatment are recommended. Positive outcomes have been shown with a treatment regimen including risk-factor modification, particularly smoking cessation and control of diabetes, exercise, and pharmacotherapy. Pentoxifylline has been used since 1984 for the treatment of IC with indifferent results. Recently, clinical trials with cilostazol, a drug approved for use in the United States, have shown significant effectiveness in IC patients, generally doubling their maximal walking distance at 24 weeks of treatment. Cilostazol has also been shown to be significantly more effective than pentoxifylline in improving pain-free and maximal walking distance. Other classes of drugs, such as platelet antiaggregants, are being studied for the treatment of IC, but little efficacy has been shown. Arterial revascularization by endovascular or surgical methods is an additional option but must be considered on an individual basis depending on severity of symptoms and disability in each patient. PMID- 11434896 TI - Comparative effects of cilostazol and other therapies for intermittent claudication. AB - Many patients with peripheral arterial disease (PAD) have intermittent claudication or problems with ambulation and mobility. Exercise and smoking cessation are primary therapies for claudication, but drug treatment may provide additional benefit. The data supporting use of pentoxifylline for claudication are weak, and pentoxifylline is not generally accepted as efficacious. Cilostazol is a new drug for the treatment of claudication. It appears to modestly benefit walking ability and it has other potentially useful effects, including inhibition of platelet aggregation and beneficial effects on serum lipids. In a randomized, prospective, double-blind trial examining walking ability in patients with PAD with moderate-to-severe claudication, cilostazol was superior to both placebo and pentoxifylline. PMID- 11434897 TI - Analysis of the cilostazol safety database. AB - Cilostazol, a type III phosphodiesterase inhibitor, was approved in the United States in 1999 for the reduction of the symptoms of intermittent claudication. This article summarizes the safety data from 8 cilostazol phase 3 controlled clinical trials, involving 2,702 patients: 1,374 receiving cilostazol, 973 assigned to placebo, and 355 taking pentoxifylline. The trials ranged from 12 to 24 weeks in duration. There were a total of 475 patient-exposure years on cilostazol, 357 patient-exposure years on placebo, and 135 patient-exposure years on pentoxifylline. Headache, diarrhea, and other gastrointestinal complaints were seen more often in cilostazol-treated than placebo-treated patients; pharyngitis and nausea were more common in pentoxifylline-treated than placebo-treated patients. Headache requiring discontinuation occurred in 1.3% of patients taking cilostazol 50 mg bid and 3.7% of those receiving cilostazol 100 mg bid, compared with 0.3% of placebo-treated patients. Discontinuations due to diarrhea, palpitations, or myocardial infarction were similar in cilostazol-, placebo-, and pentoxifylline-treated patients. The rate of serious cardiovascular events was similar in all 3 treatment groups. Myocardial infarction occurred in 1.0% of cilostazol-treated, 0.8% of placebo-treated, and 1.1% of pentoxifylline-treated patients. The incidence of stroke was 0.5% in both cilostazol- and placebo treated patients and 1.1% in pentoxifylline-treated patients. Total cardiovascular morbidity and all-cause mortality was 6.5% for cilostazol 100 mg bid, 6.3% for cilostazol 50 mg bid, and 7.7% for placebo. There were 16 deaths occurring in 0.6%, 0.5%, and 0.6% of cilostazol-, placebo-, and pentoxifylline treated patients, respectively. The evaluations showed no trend toward increased cardiovascular morbidity or mortality risk in patients receiving cilostazol. In addition, postmarketing surveillance in the United States, representing 70,430 patient-years of cilostazol exposure, has shown minimal accounts of myocardial infarction, stroke, or death. The safety profile of cilostazol in doses of 50 mg bid and 100 mg bid appears to offer an acceptable risk-benefit ratio in patients with intermittent claudication. PMID- 11434898 TI - Endovascular and surgical revascularization for patients with intermittent claudication. AB - Intermittent claudication (IC), the most common symptom of peripheral arterial disease (PAD), most often results from flow-reducing lesions in the arteries of the lower extremity that cause exercise-induced muscle ischemia. Intermittent claudication has a significant impact on quality of life and calls attention to PAD, which is secondary to systemic atherosclerosis and a major marker for cardiovascular morbidity and mortality. Most IC patients improve with a regimen that includes aggressive risk-factor modification, exercise, platelet inhibition, and pharmacotherapy to improve walking distance. Selected patients may require endovascular or surgical intervention if it can be offered with low risk. Endovascular procedures, most often percutaneous balloon angioplasty with or without stenting, are recommended for short-segment stenotic lesions in the aortoiliac and infrainguinal arterial segments. Combined platelet inhibition and endoluminal radiation are under study and may be useful to improve long-term outcome with these procedures. Percutaneous hemostatic puncture closure devices can also be used to reduce bleeding complications and allow more aggressive and immediate antithrombotic therapy, further improving results. Operative revascularization is recommended for patients with long-segment and multisegment disease, especially if obstruction is present. Aortofemoral reconstruction is associated with a low operative mortality and an 80% to 85% 5-year patency rate. Iliac reconstruction is recommended for isolated unilateral iliac arterial disease. Infrainguinal arterial reconstruction is associated with a 60% to 80% 5 year patency rate, with better outcomes noted for autogenous conduits than for prosthetic devices. Mechanical modification and pharmacotherapy with platelet inhibitors and anticoagulants improve long-term patency. PMID- 11434899 TI - Protection against ischemia: a physiological function of the renin-angiotensin system. AB - The renin-angiotensin system (RAS) is involved in a complex mechanism that serves to preserve the blood supply to organs so that they can maintain cellular function. Angiotensin II exerts this effect, independently of the blood pressure generated, through two time-related events: a fast opening of the reserve collateral circulation and a much slower response of new vessel formation or angiogenesis. This effect is observed in rats with ligation of the abdominal aorta and in gerbils with abrupt or progressive unilateral carotid artery ligation. Inhibition of the angiotensin-converting enzyme (ACE) or the angiotensin II receptor represses this effect, and it appears that it is mediated through a non-AT1 receptor site of angiotensin II. Many tumors, both benign and malignant, express renin and angiotensin. It seems that the stimulating action of angiotensin II on angiogenesis could also be involved in preserving the blood supply to tumor cells. Administration of converting enzyme inhibitors increases survival and decreases tumor size in tumor-bearing rats. These observations support the hypothesis that the RAS, directly or indirectly, is involved in situations in which the restoration of blood supply is critical for the viability of cells and that it is present not only in normal but also in pathological conditions such as tumors. In view of the ubiquitous presence of renins and angiotensins, it is also likely to be involved in other conditions, such as inflammation, arthritis, diabetic retinopathy, and retrolental fibroplasia, among others in which angiogenesis is prominent. In addition, angiotensin II could be involved, through the counterbalance of the AT1 and AT2 receptors, in the rarefaction of blood vessels as an etiologic component of essential hypertension. PMID- 11434900 TI - Effects of selected polychlorinated biphenyl (PCB) congeners on hepatic glutathione, glutathione-related enzymes, and selenium status: implications for oxidative stress. AB - Polychlorinated biphenyls (PCBs) induce drug metabolism that may lead to the bioactivation of PCBs themselves or alternatively may lead to oxidative events within the cell. The goal of the present study was to determine the influence of congeneric PCBs, selected as substrates for or inducers of drug metabolism, upon hepatic glutathione, glutathione-related enzymes, and selenium status. Male and female Sprague-Dawley rats received two i.p. injections per week of PCB 3 (4 chlorobiphenyl), PCB 28 (2,4,4'-trichlorobiphenyl), PCB 38 (3,4,5 trichlorobiphenyl), PCB 77 (3,3',4,4'-tetrachlorobiphenyl), PCB 153 (2,2',4,4',5,5'-hexachlorobiphenyl), or both PCBs 77 and 153 (100 micromol/kg/injection) and were killed at the end of 1, 2, or 3 weeks. Whole liver homogenates, hepatic cytosol, and microsomes were prepared. Both glutathione reductase and glutathione transferase activities were increased significantly in both male and female rats receiving PCB 77, an aryl hydrocarbon receptor agonist, as well as in those receiving both PCBs 77 and 153. No significant trend was observed in the levels of hepatic total glutathione. PCB 77 treatment decreased hepatic selenium-dependent glutathione peroxidase (SeGPX) activity in both male and female rats significantly. This decrease in activity following PCB 77 treatment was accompanied by a decrease in the cytosolic selenium-dependent glutathione peroxidase gene (GSPx1) transcript, as well as a decrease in hepatic total selenium levels. These data support the concept that exposure to the coplanar PCB 77 suppresses, via gene regulatory mechanisms, the cellular antioxidant enzyme SeGPX and that this decrease involves selenium. Lower halogenated PCBs that may be bioactivated to reactive oxygen species (ROS) producing metabolites, and higher halogenated PCBs that are not Ah receptor agonists, were inactive. PMID- 11434901 TI - Protection against glutamate toxicity through inhibition of the p44/42 mitogen activated protein kinase pathway in neuronally differentiated P19 cells. AB - Excessive levels of the neurotransmitter glutamate trigger excitotoxic processes in neurons that lead to cell death. N-Methyl-D-aspartate (NMDA) receptor over activation is a key excitotoxic stimulus that leads to increases in intracellular calcium and activation of downstream signaling pathways, including the p44/42 mitogen-activated protein (MAP) kinase pathway. In the present study, we have demonstrated that 1,4-diamino-2,3-dicyano-1,4-bis[2-aminophenylthio]butadiene (U0126), a potent and selective inhibitor of the p44/42 MAP kinase signaling pathway, prevents glutamate-induced death in neuronally differentiated P19 cells. In addition, we show that differentiated, but not undifferentiated, P19 cells expressed zeta1, epsilon1, and epsilon2 subunits of the NMDA receptor. Differentiated P19 cells exhibited specific NMDA receptor binding and intracellular calcium responses to glutamate that were blocked by the selective NMDA receptor antagonist [5R,10S]-[+]-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine (MK-801), but not U0126. Glutamate treatment of differentiated P19 cells triggered a rapid and sustained induction in p42 MAP kinase phosphorylation that was blocked by U0126. Pretreatment of differentiated P19 cells with U0126, but not other classes of protein kinase inhibitors, protected against glutamate-induced cell death. Post-treatment with U0126, even as late as 6 hr after glutamate application, also protected against glutamate toxicity. These results suggest that the p44/42 MAP kinase pathway may be a critical downstream signaling pathway in glutamate receptor-activated toxicity. PMID- 11434902 TI - Hepatic distribution of a phosphorothioate oligodeoxynucleotide within rodents following intravenous administration. AB - The pharmacokinetics of ISIS 1082, a 21-base heterosequence phosphorothioate oligodeoxynucleotide, were characterized within rodent whole liver, and cellular and subcellular compartments. Cross-species comparisons were performed using Sprague-Dawley rat and CD-1 mouse strains. Although whole liver oligonucleotide deposition and the proportion of drug found within parenchymal and nonparenchymal cells were similar between the two rodent species as a function of both time and dose, dramatic differences in subcellular pharmacokinetics were observed. Specifically, within murine hepatocyte nuclei, drug was observed at the 10 mg/kg dose, whereas in the rat nuclear-associated levels required the administration of 25 mg/kg. Under all experimental regimens, murine hepatic nuclear-associated drug concentrations were at least 2-fold higher than those found in rat liver cells. More detailed metabolic analysis was also performed using high performance liquid chromatography/electrospray-mass spectrometry (HPLC/ES-MS) and demonstrated that although the extent of metabolism was similar for rat and mouse, the pattern of n 1 metabolites varied as a function of both species and cell type. While rat and mouse hepatocytes and rat nonparenchymal cellular metabolites were predominantly products of 3'-exonuclease degradation, mouse nonparenchymal cells contained a majority of n-1 metabolites produced by 5'-exonucleolytic activity. Based upon these data, it would appear that subcellular oligonucleotide disposition and metabolism among rodent species are more divergent than whole organ pharmacokinetics might predict. PMID- 11434903 TI - Identification of a novel C-terminal domain involved in the negative function of the rainbow trout Ah receptor nuclear translocator protein isoform a (rtARNTa) in Ah receptor-mediated signaling. AB - Rainbow trout aryl hydrocarbon receptor (AHR) nuclear translocator isoform a (rtARNTa) has a negative function in AHR-mediated signal transduction. Previous analyses suggest that the negative function is at the level of DNA binding and may be due to the presence of 57 C-terminal amino acids that are strongly hydrophobic. To assess the negative activity of rtARNTa at the molecular level, hydrophobic-rich domains corresponding to amino acids 601-637, 601-631, and 616 631 were analyzed for the ability to affect the function of truncated rtARNT proteins in complementation and gel shift assays. Addition of the hydrophobic rich domains to these proteins reduced their ability to complement AHR-mediated signal transduction in mouse hepatoma cells by 65-95%. The decrease in function was related to a reduced ability of the AHR. rtARNT complex to bind DNA and not due to a lack of dimerization with AHR. Expression of the hydrophobic-rich domains on Gal4 proteins showed that the C-terminal domain of rtARNTa was unlikely to contain transactivation function; however, the hydrophobic domains reduced the ability of the Gal4 proteins to bind DNA. Immunoprecipitation and mutational experiments indicate that the hydrophobic-rich domains do not interact with the bHLH motif of AHR. Interestingly, immunoprecipitation experiments also revealed that the C-terminal hydrophobic-rich region of rtARNTa could oligomerize in vitro in a chimera with the Gal4 DNA binding domain. These findings indicate that the C-terminal hydrophobic amino acids are critical for the negative function of rtARNTa in AHR-mediated signaling and suggest that multiple mechanisms may be involved in the repression of DNA binding. PMID- 11434904 TI - Control of survival of proliferating L1210 cells by soluble guanylate cyclase and p44/42 mitogen-activated protein kinase modulators. AB - Intracellular signaling pathways involved in the survival of proliferating L1210 leukemia cells were investigated by using specific modulators. Among the various inhibitors tested, only 1H-[1,2,4]oxadiazole [4,3-a]quinoxalin-1-one (ODQ), a soluble guanylate cyclase (sGC) inhibitor, was found to induce a marked increase in caspase activity, which was associated with a loss of cell viability and a reduction in cGMP content. ODQ also provoked the processing of caspases-3 and -9, release of cytochrome c and, as early events, reduction of Bcl-2 content and dephosphorylation of Bad at Ser 112. Furthermore, YC-1, an sGC activator, and 8 Br-cGMP, a cell-permeant analogue of cGMP, exerted some protection against various apoptotic stimuli, such as serum deprivation or spermine accumulation. Although PD98059 (2'-amino-3'-methoxyflavone), an inhibitor of the p44/42 mitogen activated protein kinase (MAPK) pathway, did not increase basal caspase activity, and ODQ did not affect p44/42 MAPK phosphorylation significantly, phorbol myristate acetate stimulated p44/42 MAPK and reduced caspase activation induced by ODQ, serum deprivation, and spermine in a p44/42-dependent manner. SB203580 (4 (4-fluorophenyl)-2-(4-methylsulfonylphenyl)-5-(4-pyridyl)1H-imidazole), a p38 MAPK inhibitor, also partially protected against ODQ-induced apoptosis by increasing p44/42 MAPK phosphorylation. In conclusion, these results suggest that sGC may be relevant both for survival of L1210 cells under basal growing conditions and for protection against various apoptotic stimuli. p44/42 MAPK activation may also confer some protection from apoptosis, but apparently through a pathway largely independent of cGMP. PMID- 11434905 TI - Critical implication of transmembrane Phe310, possibly in conjunction with Trp279, in the rat gonadotropin-releasing hormone receptor activation. AB - The GnRH-R belongs to the superfamily of heptahelical GPCRs. A three-dimensional model of GnRH binding to its receptor predicted that Trp3 was the most deeply buried residue, potentially allowing it to interact with both Trp279, a highly conserved residue in the TMH 6 of GPCRs, and Phe310, present essentially in TMH 7 of GnRH-Rs. Replacement of Phe310 with Leu, the most common positional residue in GPCRs, induced a slightly decreased Bmax (1.6-fold) and affinity (3.8-fold); in addition, IP production was completely abolished. Similarly, replacement of Trp279 with Ser depressed the Bmax by 5.2-fold, the affinity by 2.3-fold, and totally abrogated IP production. The effect of the double mutation was not additive on binding, since the Bmax was reduced to the level of the Phe310Leu mutant, although the Kd was restored to a value not significantly different from that of the wild-type. The double mutant was also unable to induce IP production. Unexpectedly, no influence of any single or double substitution was noted on receptor internalization. These data provide evidence for the crucial role of Phe310, possibly in conjunction with Trp279, on GnRH transduction and suggest that the conformation for phospholipase C activation may not be required for GnRH R internalization. PMID- 11434906 TI - Action of the novel antioxidants 4GBE43 and 2BBE43 against lipid peroxidation. AB - The action and the effect of the newly synthesized compounds 4GBE43 [N-(1,2 diethyltetrahydro-1H-pyrazol-4-yl)-4-[(2E)-3,7-diethyl-2,6-octadienyl] oxybenzamide] and 2BBE43 [2-(benzyloxy)-N-(1,2-diethyltetrahydro-1H-pyrazol-4 yl)benzamide] against lipid peroxidation were studied. 4GBE43 and 2BBE43 quenched the ESR signal of diphenylpicrylhydrazyl (DPPH), suggesting that 4GBE43 and 2BBE43 act as scavengers of free radicals and that each compound quenched 6 free radical molecules. These compounds suppressed the oxidation of methyl linoleate emulsions and soybean phosphatidylcholine liposomes by a free radical initiator, suggesting that these compounds quench the lipid peroxyl radical. 4GBE43 and 2BBE43 also suppressed the spontaneous oxidation of rat brain homogenates. The inhibitory effect of 2BBE43 was of the same order of magnitude (IC50) as that of probucol. The IC50 of 4GBE43 was on the same order of magnitude as that of alpha tocopherol. However, 4GBE43 at 10(-4)-10(-5) M completely inhibited peroxidation, showing it to be more effective than alpha-tocopherol. These results suggest that 4GBE43 and 2BBE43 act as antioxidants by quenching the lipid peroxyl radical. PMID- 11434907 TI - Structure-activity relationships and inhibitory effects of various purine derivatives on the in vitro growth of Plasmodium falciparum. AB - The development of novel chemotherapeutic agents has become an urgent task due to the development and rapid spread of drug resistance in Plasmodium falciparum, the protozoan parasite responsible for cerebral malaria. Cyclin-dependent kinases (CDKs) are essential for the regulation of the eukaryotic cell cycle, and several enzymes of this family have been identified in P. falciparum. In recent years, a number of purine-derived kinase inhibitors have been synthesised, some of which display selective activity against CDKs. This report describes a study in which various purine derivatives were screened for in vitro antimalarial activity. The erythrocytic asexual stages of the chloroquine-resistant P. falciparum strain (FCR-3) were cultivated in vitro in the presence of the various purines, and their effect on parasite proliferation was determined by the [3H]hypoxanthine incorporation assay. Our results show considerable variation in the sensitivity of P. falciparum to the different purines, as well as a general independence from their effect on purified starfish CDK1/cyclin B activity, which has been the standard assay used to identify CDK-specific inhibitors. Two subfamilies of purines with moderate to poor activity against CDK1/cyclin B activity showed submicromolar activity against P. falciparum. Structure-activity analysis indicates that certain structural features are associated with increased activity against P. falciparum. These features can be exploited to synthesise compounds with higher activity and specificity towards P. falciparum. PMID- 11434908 TI - Molecular cloning and pharmacological characterization of the rat sigma1 receptor. AB - In an effort to further understand the pharmacology of sigma receptors, we have cloned the rat homolog of the sigma1 receptor. We isolated a cDNA clone (rs2-2) from rat brain tissue using reverse transcriptase-polymerase chain reaction (RT PCR) and 5' and 3' rapid amplification of cDNA ends (RACE) that encoded a full length sequence of 223 amino acids. The predicted protein sequence of the clone has high homology with that of the murine (93.3%), guinea pig (93.7%), and human (96%) sigma1 receptors. Northern analysis showed a major mRNA band of approximately 1.8 kb. RT-PCR revealed the presence of the mRNA in all the tissues tested, with high levels in the brain, spinal cord, liver, thymus, adrenal glands, and kidneys. When expressed in Chinese hamster ovary (CHO) cells, the level of sigma1 binding increased markedly, and the binding profile was consistent with sigma1 sites. However, measurable levels of sigma1 binding present in the cell lines before transfection made the interpretation of these results difficult. To ensure that the binding reflected the transfected protein, we tagged the receptors with a hemagglutinin (HA) epitope at the amino terminus and examined binding in immunoprecipitated receptors. Western analysis using an antisera against the HA epitope revealed a molecular weight of approximately 28 kDa, close to the predicted value. The receptor binding profile of the immunopurified receptor was consistent with that seen with traditional sigma1 binding sites. Thus, rs2-2.HA encodes a high-affinity [3H](+)-pentazocine binding site with characteristics of a rat sigma1 receptor. PMID- 11434909 TI - Inducible nitric oxide synthase in rat neutrophils: role of insulin. AB - Defective leukocyte-endothelial interactions are observed in experimental diabetes mellitus. Endogenous substances, including nitric oxide (NO), have anti inflammatory effects within the vasculature by reducing leukocyte adherence to post-capillary venules. The purpose of this study was to examine the activity and expression of NO synthase in neutrophils from alloxan-induced diabetic rats. Glycogen-elicited peritoneal neutrophils were obtained from diabetic rats and matching controls 10, 30, and 180 days after alloxan (42 mg/kg, i.v.) or saline injection. NO synthase activity was determined by the [3H]L-citrulline assay method. Expression of the enzyme was investigated by western blot analysis. Relative to controls, neutrophils obtained from diabetic rats presented a 2-fold increase in the activity of inducible NO synthase (iNOS), accompanied by an increase in the expression of the enzyme depicted by western blot. Treatment of diabetic animals with NPH insulin (2 IU/day, for 3 days) reduced both the activity and expression of iNOS to normal levels. Results presented suggest that overexpression of the inducible isoform of NO synthase by neutrophils may be responsible, at least in part, for the defects in leukocyte-endothelial interactions in diabetes mellitus. PMID- 11434910 TI - Inhibitory effect of quinolone antimicrobial and nonsteroidal anti-inflammatory drugs on a medium chain acyl-CoA synthetase. AB - The inhibitory effects of quinolone antimicrobial agents and nonsteroidal anti inflammatory drugs on purified mouse liver mitochondrial medium chain acyl-CoA synthetase catalyzing the first reaction of glycine conjugation were examined, using hexanoic acid as a substrate. Enoxacin, ofloxacin, nalidixic acid, diflunisal, salicylic acid, 2-hydroxynaphthoic acid, and 2-hydroxydodecanoic acid, which do not act as substrates, were potent inhibitors. Diflunisal, nalidixic acid, salicylic acid, 2-hydroxynaphthoic acid, and 2-hydroxydodecanoic acid inhibited competitively this medium chain acyl-CoA synthetase with K(i) values of 0.6, 12.4, 19.6, 13.4, and 15.0 microM, respectively. Enoxacin and ofloxacin inhibited this medium chain acyl-CoA synthetase in a mixed-type manner with K(i) values of 23.7 and 38.2 microM, respectively. Felbinac, which is a substrate, inhibited the activity of this medium chain acyl-CoA synthetase for hexanoic acid (IC50 = 25 microM). The concomitant presence of enoxacin and felbinac strongly inhibited this medium chain acyl-CoA synthetase. These findings indicate that medium chain acyl-CoA synthetases may be influenced by quinolone antimicrobial and nonsteroidal anti-inflammatory drugs. PMID- 11434911 TI - Intestinal absorption of rutin in free and conjugated forms. AB - Quercetin is one of the most common flavonoids in nature, occurring mainly in glycosidic forms such as rutin. Rutin has been reported to exert numerous biochemical and pharmacological activities, though information about its absorption and metabolism is scarce. The aim of this study was to investigate intestinal handling of luminally administered rutin in an isolated preparation of luminally and vascularly perfused rat small intestine. A synthetic perfusate free from blood components was used as vascular medium, with a perfluorocarbon as oxygen carrier. Luminal media consisted of a bicarbonate-buffered sodium chloride solution spiked with rutin (40.5 +/- 1.8 micromol/L). Viability was maintained during the entire perfusion; no differences between rutin and control perfusions for perfusion pressure, lactate-pyruvate ratio, oxygen uptake, and acid-base homeostasis were observed. About 10% of the administered rutin appeared at the vascular side, chiefly as free rutin (5.6%), but some rutin sulfate (2.5%) and glucuronide (2.0%) were also detected. The conjugates were preferentially absorbed to the vascular side, while only traces of the glucuronide (0.2%) were found in the luminal perfusate. Minute amounts of the rutin administered were located in the intestinal tissue (1.1%) in the form of unchanged rutin and its glucuronide and sulfate conjugates. The model used serves as a valuable tool for understanding intestinal handling of the bioactive flavonol glycoside rutin, and the obtained results confirm uptake of rutin in the rat small intestine. PMID- 11434912 TI - Differential effects of isoflurane and halothane on the induction of heat shock proteins. AB - Isoflurane is considered to be a less hepatotoxic volatile anesthetic than halothane since it not only undergoes quantitatively much less metabolism to form toxic reactive intermediates, but also preserves better hepatic blood flow. However, the biochemical basis for the reduced hepatotoxicity has not been elucidated. In this study, we examined the induction of two heat shock proteins, heat shock protein 70 (HSP70) and heme oxygenase-1 (HO-1), in the livers of rats pretreated with or without phenobarbital, followed by exposure to isoflurane or halothane under hypoxic conditions. In the phenobarbital-pretreated rats, the maximal induction of HSP70 was observed by halothane-hypoxia treatment, followed by a half-maximal induction by isoflurane-hypoxia treatment, and less than 30% induction by hypoxia treatment alone. Serum alanine aminotransferase (ALT) activity, an indicator of hepatic dysfunction, which correlated well with the extent of centrilobular necrosis, showed similar changes with increases in HSP70 mRNA. In contrast, HO-1 mRNA was induced only by treatment with halothane hypoxia. In addition, changes in the expression of HSP70 and HO-1 mRNAs were correlated with their protein expression in the liver. In non-pretreated rats, neither isoflurane-hypoxia exposure nor halothane-hypoxia exposure caused apparent hepatic injury. There was also no induction of HSP70 or HO-1 mRNA by these treatments in non-pretreated animals. These findings demonstrate that there is a significant difference in hepatic injury, and in the induction of HO-1 and HSP70 between halothane-hypoxia and isoflurane-hypoxia treatments. Isoflurane is known to be safer than halothane, which may, in part, be accounted for by the generation of less oxidative stress in the presence of isoflurane, as assessed by reduced induction of heat shock proteins compared with halothane treatment. PMID- 11434913 TI - Functional and toxicological characteristics of isolated renal mitochondria: impact of compensatory renal growth. AB - Mitochondria were isolated from renal cortical homogenates from control rats and rats that had undergone uninephrectomy and compensatory renal growth (NPX rats). Activities of selected mitochondrial processes, including key enzymes of intermediary metabolism, glutathione-dependent enzymes, and glutathione transport, were measured, and the effects of three mitochondrial toxicants were assessed to test the hypothesis that compensatory renal growth is accompanied by increases in mitochondrial metabolism and that this is associated with increased susceptibility to injury from oxidants or other mitochondrial toxicants. Activities of malic and succinic dehydrogenases were significantly higher in mitochondria from NPX rats than in mitochondria from control rats. Although the rates of state 3 respiration were significantly higher in mitochondria from NPX rats, the rates of state 4 respiration and respiratory control ratios were not different between mitochondria from control and NPX rats. Activities of glutathione redox cycle enzymes did not differ significantly between mitochondria from control and NPX rats. However, the rates of uptake of glutathione into mitochondria were approximately 2.5-fold higher in tissue from NPX rats than in tissue from control rats. Incubation of mitochondria from NPX rats with three mitochondrial toxicants [tert-butyl hydroperoxide, methyl vinyl ketone, and S (1,2-dichlorovinyl)-L-cysteine] caused greater inhibition of state 3 respiration and larger increases in malondialdehyde formation than similar incubations of mitochondria from control rats. These results indicate that mitochondria from hypertrophied renal cells are more sensitive to oxidants or mitochondrial toxicants. Baseline levels of malondialdehyde were also significantly higher in mitochondria from NPX rats, suggesting that a basal oxidant stress exists in mitochondria from hypertrophied cells. PMID- 11434916 TI - Transmembrane TNF (pro-TNF) is palmitoylated. AB - Human transmembrane tumor necrosis factor (pro-TNF) was examined for protein acylation. The cDNA encoding pro-TNF was expressed in both COS-1 cells and Sf9 cells and metabolic labeling with [(3)H]myristic or [(3)H]palmitic acid was attempted. The 17 kDa mature TNF secreted from the transfected cells was not labeled, whereas the 26 kDa pro-TNF was specifically labeled with [(3)H]palmitic acid. The [(3)H]palmitic acid labeling of pro-TNF was eliminated by treatment with hydroxylamine, indicating that the labeling was due to palmitoylation of a cysteine residue via a thioester bond. Site-directed mutagenesis of the two cysteine residues residing in the leader sequence of pro-TNF demonstrated that palmitoylation of pro-TNF occurs solely at Cys-47, located at the boundary between the transmembrane and cytoplasmic domains of pro-TNF. Thus, pro-TNF interacts with the plasma membrane via both its proteinaceous transmembrane domain and a lipid anchor. PMID- 11434917 TI - Non-invasive visualization of sperm mitochondria behavior in transgenic mice with introduced green fluorescent protein (GFP). AB - Using high sensitive polymerase chain reaction (PCR), we previously demonstrated that selective elimination of sperm mitochondrial DNA occurred during early embryogenesis in mouse. To analyze the process morphologically in more detail, a non-invasive, real-time observation of sperm mitochondria was used. Transgenic mice that express green fluorescent protein (GFP) exclusively in mitochondria (mtGFP-tg mice) were generated. The fluorescence in mtGFP-tg mice was strong and stable enough to carry out repeated observations under confocal laser scanning microscopy. In these mtGFP-tg mice it was revealed that the sperm mitochondria were selectively eliminated from egg cytoplasm during the two-cell stage of early embryogenesis. Therefore, mtGFP-tg mice should contribute to studies on sequential or repeated analysis of mitochondria. PMID- 11434918 TI - An interplay between the TOM complex and porin isoforms in the yeast Saccharomyces cerevisiae mitochondria. AB - The outer mitochondrial membrane of Saccharomyces cerevisiae contains two isoforms of mitochondrial porin, known also as the voltage-dependent anion channel. The isoform termed here porin1 displays channel-forming activity enabling metabolite transport whereas the second one, termed here porin2, does not form a channel and its function is still not clear. We have shown recently that in the absence of porin1, the channel within the protein import machinery (the TOM complex) is essential for metabolite transport across the outer membrane [Kmita and Budzinska, Biochim. Biophys. Acta 1509 (2000) 6044-6050]. Here, we report that the TOM complex channel may also serve as a supplementary pathway for metabolites in the presence of porin1 when the permeability of the latter is limited and the role of the TOM complex seems to increase when porin2 is depleted. PMID- 11434919 TI - West Nile virus-induced bax-dependent apoptosis. AB - The mechanism of cell death induced by West Nile virus (WNV), a causative agent of human febrile syndrome and encephalitis, was investigated. WNV-infected K562 and Neuro-2a cells manifested the typical features of apoptosis, including cell shrinkage, chromatin condensation and subdiploid DNA content by flow cytometry. DNA fragmentation into nucleosomal size and changes in outer cell membrane phospholipid composition were also observed in K562 cells. UV-inactivated virus failed to induce the above-mentioned characteristics, suggesting that viral replication may be required for the induction of apoptosis by WNV. Additionally, signals involved in WNV-induced apoptosis are associated with the up-regulation of bax gene expression. PMID- 11434920 TI - Alterations in expression of E2F-1 and E2F-responsive genes by RB, p53 and p21(Sdi1/WAF1/Cip1) expression. AB - RB, p53 and p21(Sdi1/WAF1/Cip1) interact in the induction of G1 arrest. We established osteosarcoma cell lines in which a tetracycline-regulatable promoter controls the induction of RB, p53 and p21. By using these cell lines, we investigated whether RB, p53 or p21 regulates, in the same manner or differently, expression and function of E2F-1 and its responsive genes. E2F-1 gene products and transcripts of the E2F-responsive genes decreased in response to RB. Similar changes occurred to p53 and p21 when RB is present. However, in the absence of RB, some of the E2F-responsive genes decreased in response to p53 but not to p21. Thus, RB is a critical component for regulating the E2F-responsive genes, while p53 alone affects only a subset of these genes. PMID- 11434921 TI - An NMR-based metabonomic approach to the investigation of coelomic fluid biochemistry in earthworms under toxic stress. AB - The endogenous metabolites of the coelomic fluid of the earthworm Eisenia veneta were characterised using high-resolution one-dimensional and two-dimensional 1H nuclear magnetic resonance spectroscopy. Signals from common organic acids, such as acetate, fumarate, malonate, malate, formate, and succinate, were identified together with adenosine and nicotinamide mononucleotide. The potential use of this information as a baseline dataset for future toxicological or physiological studies was demonstrated by a metabonomic analysis: a series of earthworms were dosed with the model compound 3-fluoro-4-nitrophenol, and toxic effects followed by multivariate analysis of the spectral data of the coelomic fluid. Relative concentrations of acetate and malonate were decreased in the dosed worms compared to the controls. PMID- 11434922 TI - Cell surface-associated chondroitin sulfate proteoglycans bind contact phase factor H-kininogen. AB - The kinin system has been recognized as a locally operating hormone system of cardiovascular cells, however, the molecular mechanisms regulating circumscribed kinin release on cell surfaces are not fully understood. In particular, the principal cell docking sites for the kinin precursor, high molecular weight kininogen (HK), are not fully explored. Here we demonstrate by enzymatic digestion, recombinant overexpression, and affinity cross-linking studies that cell surface chondroitin sulfate (CS) chains of proteoglycans (PGs) serve as major HK binding sites on platelet, fibroblast, liver, and endothelial kidney cells. In this way, CS-type PGs may contribute to a local accumulation of kinin precursors on cell surfaces and modulate circumscribed release of short-lived kinin hormones at or next to their site of action. PMID- 11434923 TI - Rat protein tyrosine phosphatase eta physically interacts with the PDZ domains of syntenin. AB - The tyrosine phosphatase r-PTPeta is able to suppress the malignant phenotype of rat thyroid tumorigenic cell lines. To identify r-PTPeta interacting proteins, a yeast two-hybrid screening was performed and an insert corresponding to the full length syntenin cDNA was isolated. It encodes a protein containing two PDZ domains that mediates the binding of syntenin to proteins such as syndecan, proTGF-alpha, beta-ephrins and neurofascin. We show that r-PTPeta is able to interact with syntenin also in mammalian cells, and although syntenin is a tyrosine-phosphorylated protein it is not a substrate of r-PTPeta. The integrity of both PDZ domains of syntenin and the carboxy-terminal region of r-PTPeta are required for the interaction between syntenin and r-PTPeta. PMID- 11434924 TI - Malaria parasite Plasmodium gallinaceum up-regulates host red blood cell channels. AB - The properties of the malaria parasite-induced permeability pathways in the host red blood cell have been a major area of interest particularly in the context of whether the pathways are host- or parasite-derived. In the present study, the whole-cell configuration of the patch-clamp technique has been used to show that, compared with normal cells, chicken red blood cells infected by Plasmodium gallinaceum exhibited a 5-40-fold larger membrane conductance, which could be further increased up to 100-fold by raising intracellular Ca(2+) levels. The increased conductance was not due to pathways with novel electrophysiological properties. Rather, the parasite increased the activity of endogenous 24 pS stretch-activated non-selective cationic (NSC) and 62 pS calcium-activated NSC channels, and, in some cases, of endogenous 255 pS anionic channels. PMID- 11434925 TI - The flavonoid baicalin inhibits superantigen-induced inflammatory cytokines and chemokines. AB - Excessive release of proinflammatory cytokines mediates the toxic effect of superantigenic staphylococcal exotoxins (SE). Baicalin, a flavone isolated from the Chinese herb Scutellaria baicalensis Georgi and used in China to treat infectious diseases, inhibited SE-stimulated T-cell proliferation (by 98%) and production of interleukin 1beta, interleukin 6, tumor necrosis factor, interferon gamma, monocyte chemotactic protein 1, macrophage inflammatory protein (MIP) 1alpha, and MIP-1beta mRNA and protein by human peripheral blood mononuclear cells. These data suggest that baicalin may be therapeutically useful for mitigating the pathogenic effects of SE by inhibiting the signaling pathways activated by superantigens. PMID- 11434926 TI - Functional analysis of antibacterial activity of Bacillus amyloliquefaciens phage endolysin against Gram-negative bacteria. AB - To analyze the antibacterial activity of Bacillus amyloliquefaciens phage endolysin, nine deletion derivatives of the endolysin were constructed. Each deletion mutant was overexpressed, purified and characterized. The catalytic domain was located on the N-terminal region and the C-terminus had an affinity with the bacterial envelope. The enzymatic activity remained in spite of the deletion of the C-terminal 116-amino acid region; however, the antibacterial activity was lost. These results indicate that antibacterial action requires both the C-terminal cell-binding and the N-terminal enzymatic activities. PMID- 11434927 TI - Isoproterenol inhibits resistin gene expression through a G(S)-protein-coupled pathway in 3T3-L1 adipocytes. AB - Resistin was recently identified as a hormone secreted by adipocytes which leads to insulin resistance in vivo and in vitro and might therefore be an important link between obesity and diabetes. To clarify the regulation of resistin gene expression, 3T3-L1 adipocytes were treated with various agents known to modulate insulin sensitivity, and resistin mRNA was measured by quantitative real-time reverse transcription-polymerase chain reaction. Interestingly, isoproterenol treatment reduced the level of resistin mRNA to 20% of non-treated control cells. This effect was dose-dependent with significant inhibition occurring at concentrations as low as 10 nM isoproterenol. Moreover, pretreatment of adipocytes with the beta-adrenergic antagonist propranolol almost completely reversed the inhibitory effect of isoproterenol, whereas addition of the alpha adrenergic antagonist phentolamine did not have any effect. Furthermore, the effect of isoproterenol could be mimicked by activation of G(S)-proteins and adenylyl cyclase. Thus, both cholera toxin and forskolin decreased resistin mRNA expression in a dose-dependent fashion by up to 90% of control levels. Taken together, these results suggest that resistin gene expression is regulated by a protein kinase A-dependent pathway in 3T3-L1 adipocytes. PMID- 11434928 TI - Cg-TIMP, an inducible tissue inhibitor of metalloproteinase from the Pacific oyster Crassostrea gigas with a potential role in wound healing and defense mechanisms(1). AB - We have cloned and characterized a cDNA encoding Cg-TIMP, the first tissue inhibitor of metalloproteinase identified in mollusks. The isolated cDNA encodes a protein of 221 residues that has a domain organization similar to that of vertebrate TIMPs including a signal sequence, and the 12 cysteines characteristic of the TIMP signature. Analysis of Cg-TIMP expression in adult oyster tissues, by Northern blot and in situ hybridization, indicates that Cg-TIMP was only expressed in hemocytes which are the key components of defense mechanisms in mollusks. We also observed that Cg-TIMP mRNA accumulated during shell damage and bacterial challenge. This pattern of expression suggests that Cg-TIMP may be an important factor in wound healing and defense mechanisms. PMID- 11434929 TI - Assignment of haem ligands and detection of electronic absorption bands of molybdenum in the di-haem periplasmic nitrate reductase of Paracoccus pantotrophus. AB - The periplasmic nitrate reductase (NAP) from Paracoccus pantotrophus is a soluble two-subunit enzyme (NapAB) that binds two c-type haems, a [4Fe-4S] cluster and a bis-molybdopterin guanine dinucleotide cofactor that catalyses the reduction of nitrate to nitrite. In the present work the NapAB complex has been studied by magneto-optical spectroscopy to probe co-ordination of both the NapB haems and the NapA active site Mo. The absorption spectrum of the NapAB complex is dominated by features from the NapB c-type cytochromes. Using a combination of electron paramagnetic resonance spectroscopy and magnetic circular dichroism it was demonstrated that both haems are low-spin with bis-histidine axial ligation. In addition, a window between 600 and 800 nm was identified in which weak absorption features that may arise from Mo could be detected. The low-temperature MCD spectrum shows oppositely signed bands in this region (peak 648 nm, trough 714 nm) which have been assigned to S-to-Mo(V) charge transfer transitions. PMID- 11434930 TI - Thermodynamic analysis of the binding of galactose and poly-N-acetyllactosamine derivatives to human galectin-3. AB - Galectin-3, with a wide tissue distribution and marked developmental regulation, provides significant insights into the progression of various disease and developmental stages. Recognized by its specificity for galactose, a detailed characterization of its sugar binding ability has been investigated by isothermal titration calorimetry. The results presented here complement well with the earlier studies utilizing hapten inhibition assays. Among the various lactose derivatives studied, A-tetrasaccharide emerged with the highest affinity for binding to galectin-3 combining site. This blood group saccharide exhibited a binding affinity 37-fold higher and a 102 kJ/mol more favorable change in enthalpy over lactose at 280 K indicating the existence of additional subsites for both the alpha1-3-linked N-acetylgalactosamine at the non-reducing end and the alpha1-2-linked L-fucosyl residue. The thermodynamic parameters evaluated for other ligands substantiate further the carbohydrate recognition domain to be part of an extended binding site. Binding thermodynamics of galectin-3 with the galactose derivatives are essentially enthalpically driven and exhibit compensatory changes in DeltaH degrees and TDeltaS owing to solvent reorganization. PMID- 11434931 TI - Ca(2+) transport into an intracellular acidic compartment of Candida parapsilosis. AB - In this report, we study Ca2+ transport in permeabilized Candida parapsilosis spheroplasts prepared by a new technique using lyticase. An intracellular non mitochondrial Ca2+ uptake pathway, insensitive to orthovanadate and sensitive to the V-H(+)-ATPase inhibitor bafilomycin A(1), nigericin and carbonyl cyanide p trifluoromethoxyphenylhydrazone was characterized. Acidification of the compartment in which Ca2+ accumulated was followed using the fluorescent dye acridine orange. Acidification was stimulated by the Ca2+ chelator EGTA and inhibited by Ca2+. These results, when added to the observation that Ca2+ induces alkalization of a cellular compartment, provide evidence for the presence of a Ca2+/nH(+) antiporter in the acid compartment membrane. Interestingly, like in acidocalcisomes of trypanosomatids, the antioxidant 3,5-dibutyl-4-hydroxytoluene inhibits the V-H(+)-ATPase. In addition, the antifungal agent ketoconazole promoted a fast alkalization of the acidic compartment. Ketoconazole effects were dose-dependent and occurred in a concentration range close to that attained in the plasma of patients treated with this drug. PMID- 11434932 TI - Adenovirus encoded cyclic nucleotide-gated channels: a new methodology for monitoring cAMP in living cells. AB - The current, static methodologies for measuring cyclic AMP (cAMP) may underestimate its regulatory properties. Here, we have exploited the Ca2+ conducting properties of cyclic nucleotide-gated (CNG) channels to measure cAMP in live cells, in response to various stimuli. We placed a mutated CNG channel with high sensitivity to cAMP in adenovirus to maximize and render facile its expression in numerous cell types. The ready, continuous nature of the readout contrasted with the traditional approach, which yielded similar static information, but lacked any continuous or interactive qualities. It seems fair to predict that this readily adopted approach will broaden the perception of cAMP signaling. PMID- 11434933 TI - Expression of membrane C1q in human monocyte-derived macrophages is developmentally regulated and enhanced by interferon-gamma. AB - The present study investigated when during "in vitro" maturation macrophages (MPhi) express membrane C1q (mC1q), and whether cell activation affects expression and function of mC1q. Although C1q mRNA was repeatedly detected in freshly isolated monocytes using reverse transcriptase-polymerase chain reaction, C1q protein was observed only in developing MPhi from day 1 to 4 on using immunodetection and flow cytometry. However, the quantity of mC1q and other MPhi membrane proteins differed strikingly in cells from different donors. We report here for the first time that CD14(+) and CD14(-) mC1q-bearing MPhi can develop, and that interferon-gamma increases mC1q display at the cell surface, and mC1q mediated phagocytosis. PMID- 11434934 TI - Deletion of the acidic-rich domain of the IL-2Rbeta chain increases receptor associated PI3K activity. AB - Interleukin-2 (IL-2) regulates the proliferation and homeostasis of lymphocytes through the coordinated activation of distinct signaling pathways. Deletion of the acidic-rich domain of the IL-2 receptor beta chain (IL-2Rbeta) prevents association of Src tyrosine kinases to the receptor, as well as IL-2-induced Akt activation. Cells bearing this deletion (BafbetaDeltaA) maintain full proliferation in response to IL-2 both in vivo and in vitro, suggesting that those pathways are dispensable for this important function of IL-2. In this study, we re-examined phosphatidylinositol-3 kinase (PI3K) activation in BafbetaDeltaA cells and found that, in BaF/3 IL-2RbetaDeltaA cells, deletion of the acidic domain induced constitutive activation of the receptor-associated PI3K activity. This, in turn, was responsible for the higher basal Akt activity observed in cells expressing this deletion. Based on these data, and since pharmacological abrogation of PI3K activity prevented IL-2-driven cell proliferation of BafbetaDeltaA cells, we conclude that the PI3K/Akt pathway is still functionally relevant in cells bearing this mutation. Moreover, we show that the PI3K-induced signals are, at least in part, responsible for c-myc expression. In conclusion, we have used this model to better identify those signals that are integral components of the molecular mechanisms responsible for IL-2-regulated cell proliferation. PMID- 11434935 TI - A rationale for lowering intraocular pressure in glaucoma. PMID- 11434936 TI - The pharmacology of bimatoprost (Lumigan). AB - Bimatoprost (Lumigan) is a pharmacologically unique and highly efficacious ocular hypotensive agent. It appears to mimic the activity of a newly discovered family of fatty acid amides, termed prostamides. One biosynthetic route to the prostamides involves anandamide as the precursor. Bimatoprost pharmacology has been extensively characterized by binding and functional studies at more than 100 drug targets, which comprise a diverse variety of receptors, ion channels, and transporters. Bimatoprost exhibited no meaningful activity at receptors known to include antiglaucoma drug targets as follows: adenosine (A(1-3)), adrenergic (alpha(1), alpha(2), beta(1), beta(2)), cannabinoid (CB(1), CB(2)), dopamine (D(1 5)), muscarinic (M(1-5)), prostanoid (DP, EP(1-4), FP, IP, TP), and serotonin (5HT(1-7)). Bimatoprost does, however, exhibit potent inherent pharmacological activity in the feline iris sphincter preparation, which is prostamide-sensitive. Bimatoprost also resembles the prostamides in that it is a potent and highly efficacious ocular hypotensive agent. A single dose of bimatoprost markedly reduces intraocular pressure in dogs and laser-induced ocular hypertensive monkeys. Decreases in intraocular pressure are well maintained for at least 24 hr post-dose. Human studies have demonstrated that systemic exposure to bimatoprost is low and that accumulation does not occur. The sclera is the preferred route of accession to the eye. The high scleral permeability coefficient Papp is a likely contributing factor to the rapid onset and long-acting ocular hypotensive profile of bimatoprost. PMID- 11434937 TI - Mechanism of action of bimatoprost (Lumigan). AB - Bimatoprost is a new ocular hypotensive agent that lowers intraocular pressure (IOP) in normal, ocular hypertensive, and glaucomatous eyes. Its mechanism of action has been studied in normal human subjects. Bimatoprost mildly stimulates the rate of aqueous humor flow during the day (13%) and at night (14%). Its ocular hypotensive action is due primarily to a 26% reduction in the tonographic resistance to outflow. Thus, bimatoprost enhances the pressure-sensitive outflow pathway. Additional beneficial effects may include an increase in the rate of flow via the pressure-insensitive outflow pathway (sometimes called the "uveoscleral outflow pathway") and a lowering of the extraocular recipient pressure (sometimes called "episcleral venous pressure"). Reduction of tonographic resistance to aqueous humor outflow reduces steady-state IOP, an effect that is beneficial for the treatment of glaucoma. In addition to its effect on steady-state IOP, reduction of resistance allows the eye to recover more quickly from transient IOP elevations. The former effect is common to all ocular hypotensive drugs, but the latter effect is an exclusive property of drugs that reduce outflow resistance, such as bimatoprost. PMID- 11434938 TI - Efficacy and safety of bimatoprost in patients with elevated intraocular pressure: a 30-day comparison with latanoprost. AB - PURPOSE: To compare the safety and efficacy of bimatoprost and latanoprost in patients with primary open-angle glaucoma or ocular hypertension. METHODS: This was a 30-day, multicenter, double-masked, randomized, clinical trial. Patients (n = 64) diagnosed with primary open-angle glaucoma or ocular hypertension were randomly assigned to receive bimatoprost 0.03%, latanoprost 0.005%, or vehicle topically in both eyes once daily, in the evening, for 29 days. The primary endpoint was the reduction in IOP from baseline on day 14 and day 29. Secondary outcome measures included eye examinations and safety parameters. RESULTS: Bimatoprost and latanoprost significantly lowered IOP from baseline (p <.001). Bimatoprost lowered IOP more than latanoprost at every timepoint measured (bimatoprost: 25-34% reduction, 5.9-8.9 mm Hg; latanoprost: 20-31% reduction, 4.4 7.9 mm Hg), although the between-group differences did not reach statistical significance. Over the 12-hour course of IOP measurements on day 29, bimatoprost provided better diurnal IOP control than latanoprost (p =.0378, area under the curve of diurnal IOP reductions, 1-way ANOVA with pairwise t-test). Both treatment regimens were safe and well tolerated, with no significant between group differences in reports of specific adverse events. The most common side effect was conjunctival hyperemia, which was similarly apparent in the bimatoprost and latanoprost treatment groups. CONCLUSIONS: At the end of this 30 day trial, once-daily bimatoprost 0.03% provided better diurnal IOP control than latanoprost and was safe and well tolerated in patients with ocular hypertension and glaucoma. PMID- 11434939 TI - Six-month comparison of bimatoprost once-daily and twice-daily with timolol twice daily in patients with elevated intraocular pressure. AB - The efficacy and safety of bimatoprost, a member of a new class of pharmacological agents called prostamides, were compared with the efficacy and safety of timolol in patients with glaucoma or ocular hypertension. Pooled 6 month results from two ongoing, multicenter, randomized, double-masked, clinical trials were analyzed. Patients were randomized in a 2:2:1 ratio to treatment with bimatoprost 0.03% once a day ([QD] n = 474), bimatoprost 0.03% twice a day ([BID] n = 483), or timolol 0.5% BID (n = 241). Scheduled visits were at prestudy, baseline, week 2, week 6, month 3, and month 6. The primary outcome measure was in diurnal intraocular pressure ([IOP] 8 AM, 10 AM, 4 PM, 8 PM). Bimatoprost QD provided significantly greater mean IOP reductions from baseline than timolol at every time of the day and at each study visit (p A transition at nt 8490, leading to an exchange of alanine 259 by threonine. The mutation was present in all family members with PROC deficiency. The carriers of the isolated PROC mutation were asymptomatic at ages of 4, 7, 10, 11, and 80 years. The combination of the PROC mutation with a PROS deficiency in two family members triggered venous thromboembolism at age 31 and 6 years, respectively. The PROS deficiency was associated with complete exclusion of one PROS allele. Two family members with isolated PROS deficiency are still asymptomatic at age 21 and 9 years, respectively. Our findings in this family suggest that the heterozygous mutation at codon 259 of the PROC gene represents a mild thrombotic risk factor and only confers a high thrombotic risk in combination with a second defect, such as the complete exclusion of one PROS allele. PMID- 11434941 TI - No Influence of acute hypertriglyceridemia on plasma t-PA in healthy male volunteers. AB - Dietary effects on liver blood flow may have biased the previously observed effects of hypertriglyceridemia on systemic tissue-type plasminogen activator (t PA) concentrations. Therefore, in this study the effects of hypertriglyceridemia on plasma t-PA were determined by inducing hypertriglyceridemia with an intravenous fat emulsion (Intralipid) infusion. In a randomised crossover fashion, eight healthy male volunteers received Intralipid 10% (1.5 ml/min) or 0.9% saline for 2 h and 45 min. After 2 h of infusion, t-PA antigen, t-PA activity, t-PA/plasminogen activator inhibitor (PAI-1) complex, and PAI-1 activity were determined. Concomitantly, the effects of Intralipid t-PA clearance were determined from steady-state t-PA antigen concentrations of a 45-min recombinant tissue-type plasminogen activator (rt-PA) infusion (31.25 microg/min). Liver blood flow was assessed from steady-state concentrations of a continuous sorbitol infusion. Differences between treatments were calculated using the prevalue as the covariate. No significant differences were observed in mean+/-S.D. endogenous concentrations of t-PA antigen, 4.5+/-0.9/4.1+/-0.9 ng/ml (Intralipid vs. saline infusion; difference of 0.3 ng/ml, 95% confidence interval, CI: -0.2, 0.8); t-PA activity, 0.69+/-0.21/0.68+/-0.21 U/ml (difference of 0.04 U/ml, CI: -0.17, 0.25); t-PA/PAI-1 complex, 2.0+/-1.3/1.6+/-1.0 ng/ml (difference of 0.1 ng/ml, CI: -0.8, 0.6); and PAI-1 activity, 7.3+/-5.1/7.1+/-5.1 U/ml (difference of 0.26 U/ml, CI: -3.7, 4.3). Mean t-PA clearance and liver blood flow were unaffected by the Intralipid infusion. These results indicate that acute hypertriglyceridemia does not alter plasma fibrinolytic parameters in healthy male volunteers. PMID- 11434942 TI - Results of a new rapid d-dimer assay (cardiac d-dimer) in the diagnosis of deep vein thrombosis. AB - The objective was to evaluate the accuracy of a new full blood rapid D-dimer assay in the diagnosis of suspected deep vein thrombosis (DVT). In 100 consecutive patients with suspected DVT, clinical probability was staged according to a pretest score proposed by Wells. For the determination of plasma D dimer, heparin and citrate blood samples were drawn, and Cardiac D-dimer, STA LIA, and Tina-quant tests were performed. Final diagnosis was confirmed either by duplex sonography or ascending venography. DVT was diagnosed in 37%, thrombophlebitis in 10%, and no venous thromboembolism was diagnosed in 52%. In 2% pulmonary embolism was detected and one patient was dismissed before final diagnosis. Cardiac D-dimer assay from citrate tubes showed a sensitivity of 88.6%, a specificity of 54%, a positive predictive value of 57.4%, and a negative predictive value of 87.1%. Nearly identical results were observed with heparin tubes. Corresponding results were 88.6%, 48%, 52.5%, and 85.7% for STA-LIA and 88.6%, 46%, 53.4%, and 85.2% for Tina-quant, respectively. In conclusion, we can say that Cardiac D-dimer is a rapid, whole blood assay with a great potential for clinical use. It can help in diagnosing DVT from citrate as well as heparin tubes with comparable sensitivity, specificity, positive and negative predictive values as STA-LIA and Tina-quant tests. PMID- 11434943 TI - Incorporation of map kinases into the platelet cytoskeleton. AB - Erk1 (p44) and erk2 (p42) mitogen-activated protein (MAP) kinases are activated in agonist-stimulated platelets, although their role(s) in the activation process is unknown. In the present study, erk1, erk2 and the phosphorylated forms of both enzymes became associated with the contractile cytoskeleton in thrombin stimulated platelets. Enzyme incorporation was accompanied by an increase in MAP kinase activity in the cytoskeleton, which was inhibited by PD98059. Pretreatment of the platelets with the arginine-glycine-aspartic acid-serine (RGDS) polypeptide enhanced both the cytoskeletal association and the enzyme activity, but cytochalasin D had no significant effect. Platelets from a patient with Glanzmann's thrombasthenia lack the alpha(IIb)beta(3) integrin and form only a rudimentary cytoskeleton, however, this cytoskeleton is enriched with both erk1 and erk2. These data suggest either that MAP kinases play a role in cytoskeletal rearrangement or that the cytoskeleton act as a frame to align MAP kinases with substrates in a highly integrated signal transduction pathway. PMID- 11434944 TI - Heparin and a cyclic octaphenol-octasulfonic acid (GL-522-Y-1) bind with high affinity to a 47-kda protein from vascular endothelial cell surface and stimulate the synthesis and structural changes of heparan sulfate proteoglycan. AB - The effect of a cyclic octaphenol-octasulfonic acid (GL-522-Y-1), upon the synthesis of a heparan sulfate proteoglycan synthesized by endothelial cells (rabbit aorta and human umbilical vein) were studied. The cells were exposed to the compounds at various concentrations for different periods of time and the synthesized heparan sulfates analyzed by a combination of agarose gel electrophoresis and enzymatic degradation. The GL-522-Y-1, like heparin, change the sulfation pattern and stimulate two- to three-fold the synthesis of heparan sulfate proteoglycan secreted by rabbit and human endothelial cells in culture. GL-522-Y-1, besides being 100 times more active than heparin, also produces a significant enhancement of cell surface heparan sulfate in human vein endothelial cells. The effect of GL-522-Y-1 is completely abolished by methylation or acetylation of its free hydroxyl groups. Both heparin and GL-522-Y-1 have high affinity for a 47-kDa protein present at the surface of endothelial cells. These and other results lead us to speculate that the antithrombotic activity of heparin and GL522 "in vivo" could be related, at least in part, to the increased production of the heparan sulfate proteoglycan by endothelial cells. PMID- 11434945 TI - Assessment of in vitro-generated platelet microparticles using a modified flow cytometric strategy. AB - Quantification of platelet microparticles (PMPs) may be a useful marker for the detection of in vivo platelet activation. Optimisation of flow cytometric methods for detection and quantification of PMPs has not been systemically evaluated. This study reports the optimisation of flow cytometric procedures for the detection of PMPs, the determination of limits of size detection using microbeads, and the characterisation of PMP generation by in vitro activation of platelets using collagen and adenosine 5' diphosphate (ADP). Fluorescent and plain microbeads proved useful for defining the limits of the flow cytometer in detecting PMPs. A systematic calibration of the forward scatter (FS) threshold parameter (size) of the flow cytometer using microbeads allowed for the detection of very small particles (down to 0.1 microm diameter). PMPs generated in vitro using ADP and collagen were reliably detected by flow cytometry using monoclonal antibodies (MAb) directed towards platelet surface membrane glycoproteins (Gp). The PMP events were detected in the FS low (i.e., small size events) and fluorescence (FL) high (i.e., platelet Gp MAb-labelled events) region. PMPs of different size profiles were observed for each of the agonists. Flow cytometry can be used as a tool in the assessment of PMPs. As detection of particles of this type is at the limit of resolution of flow cytometers, careful attention is required with the choice of platelet-specific MAb, isotype control, and optimisation of procedure setup and performance. PMID- 11434946 TI - A micromethod to measure platelet aggregation and atp release by impedance. AB - A standard electrode has been modified in order to detect platelet aggregation by impedance in small volumes of whole blood. The modified electrode was constructed to fit into a standard 0.5-ml cuvette normally employed for detecting platelet aggregation in platelet-rich plasma (PRP) on a Chrono-log Aggregometer (Haverford, PA). Whole blood samples were mixed 1:1 with saline and total volumes of 1 ml, 500 microl and 300 microl were tested. ATP release was recorded concurrently with aggregation by measuring the luminescence from the firefly luciferin-luciferase reaction. The modified electrode could detect aggregation in the 300- and 500-microl samples with the same efficiency as the 1-ml samples. ATP release in the 300- and 500-microl samples yielded equivalent levels of release as detected with the 1-ml samples. This electrode is well suited for evaluation of platelet function in research and clinical blood samples where only small volumes of blood are available. PMID- 11434947 TI - Fibrinogen naples I (B beta A68T) nonsubstrate thrombin-binding capacities. AB - Fibrinogen Naples I (Bbeta A68T) is characterized by defective thrombin binding and fibrinopeptide cleavage at the fibrinogen substrate site in the E domain. We evaluated the fibrinogen of three homozygotic members of this kindred (II.1, II.2, II.3) who have displayed thrombophilic phenotypes and two heterozygotic subjects (I.1, I.2) who were asymptomatic. Electron microscopy of Naples I fibrin networks showed relatively wide fiber bundles, probably due to slowed fibrin assembly secondary to delayed fibrinopeptide release. We evaluated 125I-thrombin binding to the fibrin from subjects I.1, I.2, II.1, and II.2 by Scatchard analysis with emphasis on the high-affinity site in the D domain of fibrin(ogen) molecules containing a gamma chain variant termed gamma'. Homozygotic subjects II.1 and II.2 showed virtually absent low-affinity binding, consistent with the Bbeta A68T mutation, whereas heterozygotes I.1 and I.2 showed only moderately reduced low-affinity binding. The homozygotes also showed impaired high-affinity thrombin binding, whereas that of the heterozygotes was nearly the same as normal. Genomic sequencing of the gamma' coding sequence (I.2, II.2), ELISA measurements of two gamma' chain epitopes (L2B, gamma'409-412, and IF10, gamma'417-427) (I.2, II.1, II.2, II.3), and mass spectrometry of Naples I fibrinogen (II.2) showed no differences from normal, thus indicating that there were no abnormal structural modifications of the gamma' chain residues in Naples I fibrinogen. However, thrombin reportedly utilizes both of its available exosites for binding to high- and low-affinity sites on normal fibrin, suggesting that binding is cooperative. Thus, reduced high-affinity thrombin binding to homozygotic Naples I fibrin may be related to the absence of low-affinity binding sites. PMID- 11434948 TI - Type IV laryngotracheoesophageal clefts -- recent 5 year experience at Great Ormond Street Hospital for Children. AB - This article summarises a 5 year experience of type IV laryngotracheoesophageal clefts from Great Ormond Street Hospital for Children, London. We present six infants who had type IV clefts, two of whom are long-term survivors, and we review the literature. The recognition of specific complicating issues may lead to a standardised approach, which can result in successful repair and long-term survival. Increased awareness of the condition and early diagnosis combined with aggressive, planned surgical intervention in centres of paediatric expertise should result in a significant reduction of current mortality. PMID- 11434949 TI - Morphological changes of vestibular ganglion cells in human fetuses and in pediatric patients. AB - The temporal bone histopathology of human vestibular ganglion cells of fetuses and pediatric patients was studied. In the first study, we traced the morphological changes in vestibular ganglion cells in human fetuses ranging from 13 weeks to 39 weeks of gestational age by using 13 temporal bone serial sections. Vestibular ganglion cells had reached histological maturity by the 24th week of gestation and the volume of vestibular ganglion cell cytoplasm increased until the 39th week of gestation. In the second study, the temporal bone serial sections of seven neonates, eight infants and five children were investigated to reveal pathological changes in vestibular ganglion cells. Morphological changes in vestibular ganglion cells in human fetuses were revealed. Vestibular ganglion cells were changed pathologically by intracranial disease and variety etiology affecting the inner ear, because these are located in the internal auditory canal between the brain and labyrinth. PMID- 11434950 TI - Doppler echocardiographic evaluation of pulmonary arterial pressure in children with allergic rhinitis. AB - In children, persistent upper airway obstruction may lead to increased pulmonary arterial pressure (PAP). Allergic rhinitis (AR) is one of the frequent cause of persisting upper airway obstruction by nasal blockage in childhood. Regular use of nasal topical corticosteroids are effective in reducing nasal blockage and obstruction. However, whether symptomatic children with AR have increased PAP and curative effect of topical steroids are not known. The aims of this study were to clarify whether children having active symptoms of AR have increased PAP and to investigate the curative effect of reducing nasal obstruction by topical corticosteroids. Twenty-three children, aged between 5 and 16, diagnosed as AR, consisted of 17 seasonal AR (SAR) and seven perennial AR (PAR), were included in the study. Nineteen age and sex matched healthy children were received as controls. PAP was measured by using Doppler echocardiography in all subjects and symptom scores of AR were recorded in rhinitis group. After first evaluation, nasal steroid, budesonid, was given to rhinitis group for three months. Mean systolic PAP was 33.4+/-3.1 for children with AR mmHg and 23.6+/-4.3 mmHg for the control group. The difference was statistically significant (P<0.05). Mean systolic PAP of children with PAR was significantly higher than children with SAR (P<0.05). In rhinitis group, mean PAP decreased significantly after relief of upper airway obstruction by nasal corticosteroid therapy to normal level of 24.9+/-3.6 mmHg (P<0.05). Our results showed that children with AR may have significantly higher PAP than healthy subjects and decreased to normal levels after relieving nasal blockage by nasal corticosteroids. Nevertheless, Doppler echocardiography is a safe, non-invasive and practical tool for cardiac investigation of children with AR. Therefore, in symptomatic period, evaluation of PAP of children with AR by using Doppler echocardiography may be useful in the planning and following of their therapy. PMID- 11434951 TI - Generalisability of clinical trials in otitis media with effusion. AB - OBJECTIVE: The impact of a randomised controlled trial (RCT) upon practice depends on its external validity (generalisability). This paper summarises and illustrates a framework for judging and augmenting external validity, emphasising its application to treatment trials in otitis media with effusion (OME) so as to permit stronger inferences in the future. METHODS: The external validity of two surgical trials in the field of OME (TARGET, UK and KNOOP-3, the Netherlands) has been examined within a framework emphasising effect modification, in four specific ways: (1) comparison of the demographic characteristics of the trial population with the domain population; (2) studying the distributions on possible effect modifiers (i.e. variables conditioning the benefit from intervention); (3) studying whether effect modification occurs in the analyses; and (4) comparing outcome measures between the randomised and the eligible but non-randomised children. RESULTS: For neither KNOOP-3 and TARGET were large discrepancies found between randomised and non-randomised children for any of the demographic variables. Differences in distributions along possible effect modifiers were found, but the overlaps were large enough for it still to be possible to study whether these factors indeed modified the outcome. Results for the randomised and non-randomised but eligible patients were similar. The results of both trials therefore appear to be generalisable to their domain populations. CONCLUSIONS: A superficial contrast in the results (KNOOP null, TARGET positive) does not amount to a contradiction, because of differences in the clinical question appropriate to the respective age and populations defined. Attention to quality of design and external validity of randomised controlled trials should achieve higher applicability. PMID- 11434952 TI - The role of postoperative chest radiography in pediatric tracheotomy. AB - A postoperative chest radiograph has traditionally been obtained after tracheotomies to evaluate for the presence of a pneumothorax and to assess tube position. Several recent studies in adults have questioned the usefulness of routine postoperative chest radiography in uncomplicated cases, but the role of post-operative chest radiography in pediatric patients has not been previously reviewed. We performed this study to examine the clinical utility of post tracheotomy chest radiography in pediatric patients and determine if this routine practice impacts patient management enough to merit continued usage. A retrospective review was performed of 200 consecutive pediatric patients who underwent tracheotomies by the otolaryngology service in a tertiary care pediatric hospital from January 1994 to June 1999. All patients received postoperative chest radiographs. Five of 200 patients had a new postoperative radiographic finding, with three requiring interventions. Two patients required chest tube placement for pneumothorax, and one patient required tracheostomy tube change for repositioning. Fifty-one patients, including both pneumothoraces, exhibited clinical signs of pneumothorax (decreased breath sounds or oxygen saturation) in the immediate postoperative period. Chest X-ray ruled out a pneumothorax in the remaining 49 patients. The majority of these 51 patients were less than 2 years old (94%, P=0.002) or weighed less than 17 kg (89%, P=0.004). Postoperative chest X-rays yielded clinically relevant information in 168 patients that fell into one or more of four high risk categories: age less than 2, weight less than 17 kg, emergent procedures, or concomitant central line placement. Avoiding chest X-rays in the remaining 32 patients would have resulted in potential savings of $5000, which does not reflect the actuarial cost of a missed complication. Since the majority of our patients (84%) fell into a high risk category, we feel it would be prudent to continue obtaining postoperative chest radiographs following all pediatric tracheotomies. PMID- 11434953 TI - Bacteriological and PCR analysis of clinical material aspirated from otitis media with effusions. AB - Otitis media with effusions (OME) can lead to significant hearing loss in childhood. Although previous studies have shown that bacterial DNA is present in a significant percentage of effusions sterile by culture, whether the DNA represents viable organisms or 'fossilized remains' is unknown. The aim of the present study was the determination of Streptococcus pneumonia, Moraxella catarrhalis and Haemophilus influenza in the clinical materials from OME. For this purpose, effusion samples aspirated from the mid-ear were analyzed bacteriologically. Samples were also tested with polymerase chain reaction (PCR) assay. A total of 37 samples obtained from 20 patients aging between 4 and 14 were analyzed. In 17 patients, both ears demonstrated effusions, whereas in three patients, only one ear had effusions. In aerobic bacterial culture assays, nine samples (24.3%) showed bacterial growth. None of the samples were positive for anaerobic culture experiments. PCR analysis of the samples showed that 35 samples (94.5%) contained bacterial DNA. In 19 samples, only one bacterial species and in 16 samples more than one bacterial species were detected. The results of this study demonstrate that effusion fluid from otitis media cases contain a battery of bacterial species and these bacteria might play roles in the pathogens of OME. Our results indicate the PCR technique is more specific and sensitive in detection of bacteria in middle-ear effusion of OME, compared with conventional methods. PMID- 11434954 TI - Concentration of nitric oxide metabolites in middle ear effusion. AB - Free radicals such as nitric oxide (NO) seem to be important in the pathogenesis of otitis media with effusion (OME). NO can be quantitated by measuring its metabolites, nitrate (NO(3)(-)) and nitrite (NO(2)(-)). The purpose of this study is to determine the concentrations of NO in human middle ear effusion (MEE). Samples of human MEE were collected at the time of myringotomy and tympanostomy tube insertions. The type of MEE was classified as serous (SOM), mucoid (MOM) or purulent (POM) at the time of surgery. Samples of MEE were assayed for NO metabolites (nitrate and nitrite) with colorimetric assay (Griess method). Concentrations of NO metabolites were highest in MOM followed by SOM and POM. This study suggests that NO is present in human MEE and may play an important role in the pathogenesis of OME. PMID- 11434955 TI - Endoscopic division of the ary-epiglottic folds in severe laryngomalacia. AB - OBJECTIVE: Laryngomalacia is the commonest cause of congenital stridor. The underlying anatomical abnormality associated with this condition is a prolapse of the supraglottic tissues into the laryngeal inlet during inspiration, and may involve the epiglottis, ary-epiglottic folds and the corniculate mounds of the arytenoids. However, it has been noted that the most consistent structural abnormality seen in these cases is the shortening of the ary-epiglottic folds and marked side to side curling of the epiglottis. We describe the follow-up and outcome of 33 cases treated by the simple division of the ary-epiglottic folds. METHOD: All case notes were reviewed with respect to indications of operation, age of operation, endoscopic findings, operative technique, complications and follow-up until resolution of symptoms. RESULTS: Surgical outcomes could only be ascertained in only 32 patients. Twenty-two cases (68.7%) showed complete resolution of stridor and associated complications of laryngomalacia. In the remaining ten cases who could be followed up (31.2%), seven patients (21.8%) showed partial resolution with no further surgery required, two patients (6.2%) required additional excision of redundant mucosa as second procedure, one patient (3%) with associated cleft lip/palate and tracheomalacia had to undergo a tracheostomy. There was improvement in feeding after surgery in all the 12 patients (100%) who had had pre-operative feeding difficulties. All the four patients with cyanosis pre-operatively were cured, but one of the two patients with apnoeic episodes pre-operatively continued to have apnoeic spells despite resolution of their laryngomalacia. CONCLUSION: Simple endoscopic excision of the ary-epiglottic folds is a quick, reliable, highly effective procedure with very few complications. We recommend its use as the first line option in the management of severe laryngomalacia, with more extensive methods reserved for the very occasional case of primary failure. PMID- 11434956 TI - Hearing results of canal wall reconstruction tympanoplasty for middle ear cholesteatoma in children. AB - OBJECTIVE: To investigate post-operative hearing results in children with middle ear cholesteatoma, and to analyze the correlation between hearing results and clinical factors and findings before and during the operation. PATIENTS AND METHODS: One hundred and twenty-four ears of 123 children were operated on for middle ear cholesteatoma at the age of 10 years or younger by canal wall reconstruction tympanoplasty and were followed up more than 1 year after the final operation. We evaluated the average air and bone conduction hearing levels at the speech ranges before the first operation (pre-operative hearing) and after the final operation (post-operative hearing). RESULTS: The mean of the average air conduction hearing level of 124 ears was significantly improved from 34.7 to 27.1 dB after the final operation. Among them, 84 ears (67.8%) showed a hearing level of 30 dB or less post-operatively. Post-operative hearing was better in the one-stage group than in the staged group. However, more than one-half of the ears which underwent type IV tympanoplasty in the staged group showed post-operative air conduction hearing level of < or =30 dB. Significant improvement in post operative hearing was noted in ears with normal middle ear mucosa or middle ear effusion at the final operation. No correlation between hearing improvement and clinical factors such as age, type of cholesteatoma or presence of otitis media with effusion at the first operation was found. CONCLUSIONS: Children with middle ear cholesteatoma at the age of 10 years or younger exhibited good hearing post operatively. Satisfactory hearing improvement is expected even in ears without the superstructure of the stapes if staged tympanoplasty is conducted. Canal wall reconstruction tympanoplasty for pediatric cholesteatoma was successful in terms of hearing results and the success was unrelated to various clinical factors. PMID- 11434957 TI - Temporal bone histopathological study of Noonan syndrome. AB - Four temporal bone specimens, obtained from three individuals 1--6 years of age with Noonan syndrome (NS), were studied histopathologically. All four specimens were accompanied by similar inner ear abnormalities including the reduced number of spiral ganglion cells, enlarged lateral semicircular canal, and dislocated endolymphatic sac and vestibular aqueduct. The mean population of spiral ganglion cells (15,699 cells) was approximately half of those (32,978 cells) in four age matched control cases. In addition, they had several middle ear abnormalities including the remaining mesenchyme and dehiscence of the facial canal. To our knowledge, this is the first report to describe the histopathological temporal bone findings in patients with NS. We discuss the implications of the observed abnormalities with regard to clinical issues. PMID- 11434958 TI - Congenital cystadenoma of the tongue in a neonate case report with review of literature. AB - We present a congenital cystadenoma of the tongue in a neonate, which presented at birth. Cystadenomas are uncommon tumors that form from salivary gland duct tissue and are more commonly seen in adults. This is the youngest case to be reported in the English literature. A review of literature with differential diagnosis and management is presented. PMID- 11434959 TI - Manipulating cattle distribution with salt and water in large arid-land pastures: a GPS/GIS assessment. AB - Several of the problems associated with grazing animals in extensive settings are related to their uneven patterns of use across the landscape. After fencing, water and salt are two of the most frequently used tools for affecting cattle distribution in extensive settings. Cattle are attracted to water in arid regions, but mixed results have been obtained with salt and mineral supplements. The goal of this study was to evaluate the efficacies of salt and water manipulations for affecting cattle distribution in large (>825ha) pastures. This was accomplished by fitting cattle with global positioning system (GPS) collars and monitoring their travels and activities in a three pasture, Latin-square design where water and salt shared a common location and water or salt were moved individually to distant areas. Mean distance of cattle from water (&xmacr;=1.16km) was unaffected by treatments (P=0.79) suggesting that cattle followed movements of water tanks. Distance traveled daily (&xmacr;=5.78km), time devoted to grazing (11.0h per day), time devoted to resting (10.1h per day), and the area (&xmacr;=325ha) of minimum convex polygons were also unaffected by treatment implying that cattle did not compensate for separated water and salting areas with increased travels or disruptions of habitual grazing and resting activities. Centers of activity for cattle shifted further (P=0.02) when water (&xmacr;=1.49km) was moved than when salt (&xmacr;=1.00km) was relocated. Mean distance of cattle from salt increased from 1.03km, when salt and water were together, to 1.73km (P=0.08) when salt and water were separated. This implied that cattle made less effort to remain near salt. Also, when water and salt were separated, cattle were found within 250m of water 354 times and close to salt only 38 times. Movement of drinking water to distant points in pastures was the most effective tool for altering cattle distribution. When cattle and salt were introduced to a new portion of a pasture, cattle used the new area for about 2 days, and then began drifting back toward previously used portions of the pasture. Manipulations of salting stations will not significantly rectify serious livestock distribution problems in extensive arid-land pastures. PMID- 11434960 TI - A note on the influence of starting position, time of testing and test order on the backtest in pigs. AB - The backtest determines the coping behaviour of a piglet in a standardised stress situation, which might be a measure for the coping style of that animal. Backtest results are related to other parameters such as immune responses and production. In this study, the backtest was standardised and it was studied if time of testing or the order in which animals were tested influenced backtest results. One hundred and eight-four piglets from 18 sows were tested at 3, 10 and 17 days of age. Before testing, the sow was disturbed to ensure that all piglets were awake. This standardisation of the starting position before testing did not improve the correlations between backtest results in the successive tests. No relations were found between backtest results and time of testing or test order. PMID- 11434961 TI - PGF(2alpha)-induced nest building and choice behaviour in female domestic pigs. AB - The domestic pig, Sus scrofa, builds a maternal nest in the day before parturition. A model for porcine nest building has been established, in which exogenously administered prostaglandin (PG)F(2alpha) is used to induce nesting behaviour in cyclic, pseudopregnant and pregnant pigs. This experiment was designed to examine the effect of PGF(2alpha) on the preferences of non-pregnant gilts for pens bedded with straw compared with bare pens. Ten 6-month-old nulliparous female pigs (gilts) were tested in an arena, which consisted of four pens (1.8mx1.7m), a neutral area (1.5mx3.4m) and a start area (1.5mx3.4m). Two of the pens contained 2kg of fresh straw and the remainder of the testing arena was devoid of straw. On the first day of testing half of the pigs were given a control intramuscular injection of 3ml 0.9% saline and the remainder were given an intramuscular injection of 15mg PGF(2alpha) and their behaviour scored for 1h after treatment. On the following day the treatments were reversed, such that each pig was given both treatments (saline or PGF(2alpha)). There was no significant effect of the order of treatment on behaviour. After saline-treatment the pigs spent most of their time in the pens containing straw (59%) and the least amount of time in bare pens (5%). In the straw pens, saline-treatment induced bouts of oronasal contact with straw of a relatively long duration (11 100s), which we interpret as foraging. In the hour after PGF(2alpha)-treatment the pigs also spent most of their time in the pens containing straw (44%) and the least amount of time in bare pens (10%), but they interacted with the straw in a markedly different way. PGF(2alpha)-treated pigs displayed bouts of oronasal contact with straw of a relatively short duration (2-10s) which, together with high frequencies of pawing at straw, lifting and carrying straw in the mouth, we interpret as nest building behaviour. Superimposed on this is the finding that gilts spend more time in the neutral areas after PGF(2alpha)-treatment than they did after saline-treatment. PGF(2alpha)-treated pigs spent most of their time engaged in nesting behaviour within the straw pens but they also gathered and deposited straw in different areas of the test arena (neutral and start areas); behaviours not seen after saline-treatment. We conclude that pigs generally prefer a pen containing straw bedding to a bare pen but that PGF(2alpha) alters the way they interact with straw, inducing behaviour similar to prepartum nest building. PMID- 11434962 TI - A note on the effects of forward and rear-facing orientations on movement of horses during transport. AB - Several studies have attempted to determine the effects of orientation on a horse's ability to maintain balance during transportation. The results have often been contradictory because of differences in trailer design and lack of simultaneous comparisons. In this study, three replications of two forward-facing and two rear-facing horses were transported at the same time over a standardized course to allow for simultaneous comparisons. Each animal's total forward and backward motion during transport was calculated to estimate the effect of orientation on the horses' ability to maintain balance. The course consisted of four laps around a 3.6-km rectangular course, each lap reversing direction, totaling 14.4km. To mimic realistic travel, the course had artificial bumps, three turns (90 degrees, 45 degrees, and 135 degrees ), five straight-aways, and a hard stop at the end of each lap. Four horses were transported over the course in 3.7mx2.4m stalls mounted in-line on a 16-m long commercial straight-deck trailer (16 wheels). At the end of the first run (four laps), the orientation of each horse, two forward and two rear-facing, was reversed for a second run. Twelve horses were transported in each of the two orientations. Movement was recorded using video cameras positioned perpendicular to the horse's side. Movement while forward-facing ranged from 4.75 to 34.48m, averaging 12.95m; when rear-facing, movement ranged from 8.13 to 35.21m, averaging 16.99m, and was not statistically influenced by orientation (P=0.1219) due to high variation. Certain horses did demonstrate a superior ability to maintain balance in a particular orientation. Thus individual characteristics and other factors may play a larger role than orientation alone in the ability of horses to maintain balance during transport. PMID- 11434963 TI - Behavioural responses of red deer to fences of five different designs. AB - Capercaillie, a large species of grouse, are sometimes killed when they fly into high-tensile deer fences. A fence design which is lower or has a less rigid top section than conventional designs would reduce bird deaths, but such fences would still have to be deer-proof. The short-term behavioural responses of farmed red deer (Cervus elaphus) to fences of five designs, including four that were designed to be less damaging to capercaillie, were measured. Five deer were located on one side of a fence with a larger group (20 animals), from which they had been recently separated, on the other. The efficacy of fences in preventing deer from the small group from rejoining the larger group was also recorded. In addition to a conventional deer fence (C) the four new designs were, an inverted "L" shape (L), a fence with offset electric wire (E), a double fence (D) and a fence with four webbing tapes above (W). Four replicate groups of deer were each tested for 3 days with each fence design. Deer paced the test fence line relatively frequently (a proportion of 0.09 scan observations overall) but significantly less when deer were separated by fences E or C compared to L, W or D (overall difference between fence types, P<0.001). Deer separated by fence E spent significantly more time pacing perimeter fences than deer separated by fences of other types (overall difference between fence types, P<0.01) but deer separated by fence C maintained a low level of fence pacing overall. Analysis of behaviour patterns across the first day and the 3 days of exposure suggested that the novelty of the test fences, rather than the designs per se, influenced the behaviour of the deer. Over the course of the study, no deer crossed either C or L. Three deer crossed E and two deer crossed both W and D. On this basis, field testing, particularly of fence L, would be a useful next step. PMID- 11434964 TI - The effectiveness of a citronella spray collar in reducing certain forms of barking in dogs. AB - This study examined the effectiveness of a citronella spay collar in reducing barking in 30 dogs which wore the collar continuously, i.e. every day for 30min, or intermittently, i.e. every other day for 30min, for a period of 3 weeks. Owners rated the frequency of their dog's barking on a scale ranging from 1 (very infrequent) to 5 (very frequent) before the study began (pre-treatment condition) and at the end of every week that the collar was worn (treatment condition). A further evaluation was made at the end of week 4, following 7 days of collar deprivation (post-treatment condition). Dogs exhibited a significantly lower frequency of barking during the treatment and post-treatment conditions compared to the pre-treatment condition. Barking was most effectively reduced when dogs wore the collar intermittently. However, barking increased over the period of time the collar was worn, particularly in those dogs which wore the device every day. Barking continued to increase when the dogs stopped wearing the collar, albeit at frequencies lower than pre-treatment, particularly in those animals which had worn the collar continuously. The collar was more effective at reducing travel related barking than television or traffic related barking. Findings indicate that dogs generally habituate to the citronella spray collar irrespective of how the animal is exposed to the device. A longer period of efficacy occurs, however, if the collar is worn intermittently. This may appeal to owners who are concerned about their pet having to wear the collar for long periods of time. PMID- 11434965 TI - The involvement of dopamine (DA) and serotonin (5-HT) in stress-induced stereotypies in bank voles (Clethrionomys glareolus). AB - In order to clarify the dependency of stress-induced stereotypies on dopamine (DA) and serotonin (5-HT) functioning, undisturbed and acutely stressed stereotyping bank voles were treated during 3 weeks with the commonly used human atypical neurolepticum clozapine and the SSRI antidepressant citalopram. Clozapine blocks DA receptors (D sub (4)) and acts as a partial 5-HT antagonist (5-HT sub (2) receptors), while citalopram increases 5-HT transmitter activity. Levels of stereotypies were quantified under undisturbed conditions during the treatment period and immediately after the acute stress of handling and injections. It was demonstrated that stereotypies are markedly increased after acute stress and that citalopram effectively mitigates this stress effect. Stereotypies under undisturbed conditions were left unaffected by both clozapine and citalopram treatments.It is suggested that stress-induced increases in stereotypies are more dependent on 5-HT than DA functioning. PMID- 11434966 TI - Coronavirus derived expression systems. AB - Both helper dependent expression systems, based on two components, and single genomes constructed by targeted recombination, or by using infectious cDNA clones, have been developed. The sequences that regulate transcription have been characterized mainly using helper dependent expression systems and it will now be possible to validate them using single genomes. The genome of coronaviruses has been engineered by modification of the infectious cDNA leading to an efficient (>20 microg ml(-1)) and stable (>20 passages) expression of the foreign gene. The possibility of engineering the tissue and species tropism to target expression to different organs and animal species, including humans, increases the potential of coronaviruses as vectors. Thus, coronaviruses are promising virus vectors for vaccine development and, possibly, for gene therapy. PMID- 11434967 TI - A biochemically structured model for Saccharomyces cerevisiae. AB - A biochemically structured model for the aerobic growth of Saccharomyces cerevisiae on glucose and ethanol is presented. The model focuses on the pyruvate and acetaldehyde branch points where overflow metabolism occurs when the growth changes from oxidative to oxido-reductive. The model is designed to describe the onset of aerobic alcoholic fermentation during steady-state as well as under dynamical conditions, by triggering an increase in the glycolytic flux using a key signalling component which is assumed to be closely related to acetaldehyde. An investigation of the modelled process dynamics in a continuous cultivation revealed multiple steady states in a region of dilution rates around the transition between oxidative and oxido-reductive growth. A bifurcation analysis using the two external variables, the dilution rate, D, and the inlet concentration of glucose, S(f), as parameters, showed that a fold bifurcation occurs close to the critical dilution rate resulting in multiple steady-states. The region of dilution rates within which multiple steady states may occur depends strongly on the substrate feed concentration. Consequently a single steady state may prevail at low feed concentrations, whereas multiple steady states may occur over a relatively wide range of dilution rates at higher feed concentrations. PMID- 11434968 TI - Estimation of kinetic parameters in a structured yeast model using regularisation. AB - In this work, a procedure for estimating kinetic parameters in biochemically structured models was developed. The approach is applicable when the structure of a kinetic model has been set up and the kinetic parameters should be estimated. The procedure consists of five steps. First, initial values were found in or calculated from literature. Hereafter using sensitivity analysis the most sensitive parameters were identified. In the third step physiological knowledge was combined with the parameter sensitivities to manually tune the most sensitive parameters. In step four, a global optimisation routine was applied for simultaneous estimation of the most sensitive parameters identified during the sensitivity analysis. Regularisation was included in the simultaneous estimation to reduce the effect of insensitive parameters. Finally, confidence intervals for the estimated parameters were calculated. This parameter estimation approach was demonstrated on a biochemically structured yeast model containing 11 reactions and 37 kinetic constants as a case study. PMID- 11434969 TI - Biomass production and studies on Rhodopseudomonas palustris grown in an outdoor, temperature controlled, underwater tubular photobioreactor. AB - A temperature controlled underwater tubular photobioreactor was studied for 6 months in outdoor conditions to determine biomass production of Rhodopseudomonas palustris 42OL. Biomass output rate was very high during the study period. In July, productivity was about 75.0 g reactor(-1) d(-1) for a short period (5 days). The biomass yield averaged 0.7 g biomass dry weight (d.w.)xg acetic acid( 1). We also obtained a mean photosynthetic efficiency of 7.6% in winter and 7.1% in summer. Biomasses rich in pigments and a biodegradable thermoplastic polymer [poly-beta-hydroxybutyrate (PHB)] were obtained. During the summer, PHB was about 4.0% of the biomass d.w. The bacteriochlorophyll/carotenoid ratio was lower at midday than either in the morning or in the evening. The reduction in this ratio may have been a response of the Rhodopseudomonas cells to high irradiance (about 900 W m(-2)). A reduced rate of protein synthesis continued even in the dark. PHB and carbohydrates, synthesized in excess during the day, were used as substrates for night protein synthesis. PMID- 11434970 TI - The use of a second reporter plasmid as an internal standard to normalize luciferase activity in transient transfection experiments may lead to a systematic error. AB - beta-galactosidase reporter plasmids containing different viral or minimal promoters are commonly used to correct variable transfection efficiencies in transient transfection experiments. The transcriptional activity of these promoters is thought to be stable under most circumstances. To determine if expression of beta-galactosidase from the commonly used beta-galactosidase plasmids remains stable upon stimulation of the cells with agonists we performed transient transfection experiments. CHO cells stably expressing the rat AT(1A) receptor were transfected with RSVbeta- or CMVbeta- or pTKbeta plasmids alone or together with a reporter construct in which luciferase transcription is driven by the c-fos promoter. Luciferase and/or beta-galactosidase activity was measured from the lysate of cells treated with angiotensin II or serum. We found that agonists increased the transcriptional activity of the different beta galactosidase plasmids. The effect of angiotensin II and serum was different on the different promoters. Finally, cotransfection of other plasmids also modulated beta-galactosidase activity. These agonist induced variations of beta galactosidase activity may influence the analysis and interpretation of the results in a systematic manner. Consequently we conclude that the use of a second reporter system to control for transfection efficiency in certain types of experiments may lead to a systematic error and is questionable as a general procedure. PMID- 11434971 TI - Optimization of astaxanthin production by Phaffia rhodozyma through factorial design and response surface methodology. AB - Sequential methodology based on the application of three types of experimental designs was used to optimize the astaxanthin production of the mutant strain 25-2 of Phaffia rhodozyma in shake flask cultures. The first design employed was a factorial design 2(5), where the factors studied were: pH, temperature, percent of inoculum, carbon and nitrogen concentrations, each one at two levels. This design was performed in two medium types: rich YM medium and minimal medium, based on date juice (Yucca medium). With this first design the most important factors were determined (carbon concentration and temperature) that were used in the second experimental strategy: the method of steepest ascent was applied in order to rapidly approach the optimum. Finally, a second-order response surface design was applied using temperature and carbon concentration as factors. The optimal conditions stimulating the highest astaxanthin production were: 19.7 degrees C temperature; 11.25 g l(-1) carbon concentration; 6.0 pH; 5% inoculum and 0.5 g l(-1) nitrogen concentration. Under these conditions the astaxanthin production was 8100 microg l(-1), 92% higher than the production under the initial conditions. PMID- 11434972 TI - Improvement of a mediator-type biochemical oxygen demand sensor for on-site measurement. AB - We characterized a mediator-type biochemical oxygen demand (BOD) sensor with a three-electrode system using potassium ferricyanide (FC) and Pseudomonas fluorescens in our previous study. In the present study, we have utilized the advantages of a mediator-type biosensor, which does not require air-supply equipment for on-site measurements, and made a fully disposable sensor tip for a portable device. The tip consists of a two-electrode system with P. fluorescens immobilized on a cellulose acetate membrane and is packaged in polyester film to prevent it from drying out. By aeration with a 0.1 M NaCl solution of P. fluorescens (after growth), the sensor responses as well as their reproducibility and stability have been successfully improved. The responses increased more than seven times, and the calibration curve from 15 to 260 mg l(-1) also remained linear although the response decreased approximately half the original after at least 35 days in storage. The reproducibility of the sensor responses improved to 12.7% (average of relative standard deviations (RSDs)) in the calibration curve obtained by using the Organization for Economic Cooperation and Development synthetic sewage. Examination of real samples from three different sources showed that the BOD as determined by the sensor correlates well with the conventional 5 day BOD method (r(2)=0.982, 0.823, and 0.809). Consequently, the aeration process makes it possible to realize rapid, and in situ measurements without the long conditioning process that is generally required to activate the microorganisms immobilized on bio-films before use. Finally, we have designed a portable device that utilizes our disposable sensor tip. PMID- 11434973 TI - Antifungal activity of Artemisia annua endophyte cultures against phytopathogenic fungi. AB - Artemisia annua, well recognized for its production of antimalarial drug artemisinin, is seldom attacked by any of phytopathogenic fungi, which could be partially associated with the presence of endophytes. Present investigation is aiming at disclosing whether the endophytes inside A. annua produce antifungal substances. A total of 39 endophytes were isolated and fermented, and the ferment broth was evaluated in vitro for the antifungal activity against crop-threatening fungi Gaeumannomyces graminis var. tritici, Rhizoctonia cerealis, Helminthosporium sativum, Fusarium graminearum, Gerlachia nivalis and Phytophthora capsici. These plant pathogens are still causing wheat take-all, sharp eyespot, common rot, scab, snow mould, and pepper phytophthora blight, respectively. Out of 39 endophytes investigated, 21 can produce in vitro substances that are inhibitory to all or a few of the tested phytopathogens whereas the rest yielded nothing active. Moreover, the most active broth of endophyte IV403 was extracted with EtOAc and n-butanol, and comparisons of the antifungal activity of the extracts indicated that the major active metabolites were EtOAc-extractable. PMID- 11434974 TI - Distribution of serotonin, its metabolites and 5-HT transporters in the neostriatum of Lurcher and weaver mutant mice. AB - Serotonin (5-HT) uptake sites, or transporters, were measured in the neostriatum (caudate putamen) of wild type (+/+) mice and heterozygous (wv/+) and homozygous (wv/wv) weaver, as well as in heterozygous Lurcher (Lc/+) mutants. These topological surveys were carried out by quantitative ligand binding autoradiography using the uptake site antagonist [3H]-citalopram as a probe of innervation densities in four quadrants of the rostral neostriatum and in two halves of the caudal neostriatum. In addition, tissue concentrations of 5-HT, 5 hydroxyindole-3-acetic acid and 5-hydroxytryptophol were measured by high performance liquid chromatography with electrochemical detection in these neostriatal divisions. In +/+ mice and in Lc/+ mutants there was a dorso-ventral gradient of increasing 5-HT levels, and they exhibited a similar heterogeneity of [3H]-citalopram labeling. In contrast, the gradients of 5-HT concentrations and [3H]-citalopram binding disappeared in the weaver mutants, suggesting a rearrangement of the 5-HT innervation. This reorganization of the 5-HT system in the neostriatum was more obvious in the wv/wv and is compatible with the hypothesis that the postnatal dopaminergic deficiencies that characterize weaver mutants lead to a sprouting of fibers and thus constitute a genetic model of dopaminergic denervation that leads to a 5-HT hyperinnervation. PMID- 11434975 TI - Higher order chromatin degradation in glial cells: the role of calcium. AB - Higher order chromatin degradation (HOCD), i.e. the scission of nuclear chromatin loops at the matrix attachment regions (MARs), is a hallmark of programmed cell death. We have previously demonstrated that hydrogen peroxide (H(2)O(2)) induces rapid HOCD in cultured oligodendrocytes generating two subpopulations of DNA fragments of >or=400 and 50-200 kb. In the present study, we examined the involvement of calcium in this process. HOCD was induced in primary rat oligodendrocytes by exposure to 1 mM H(2)O(2) and assessed by field inversion gel electrophoresis with and without S1 endonuclease digestion, to detect single and double stranded fragmentation, respectively. Chelating intracellular calcium with BAPTA/AM prior to H(2)O(2) exposure inhibited HOCD in a dose-dependent manner. Complete inhibition of HOCD was attained with 50 muM BAPTA/AM. The pretreatment of cells with desferroxamine mesylate, which may lower intracellular calcium levels, also resulted in a profound inhibition of HOCD, but the initial chromatin digestion into >or=400 kb single stranded DNA fragments was unaffected. Neither removing extracellular calcium nor blocking calcium release from intracellular stores with TMB-8 affected HOCD. Moreover, increasing intracellular calcium with A23187 calcium ionophore did not induce HOCD. Subsequent study in nuclei purified from C6 glioma cells revealed that the endonuclease responsible for HOCD is calcium-independent, but is magnesium-dependent. Magnesium-induced HOCD was not affected by the removal of calcium from nuclei with EGTA, but was practically abrogated in nuclei prepared from BAPTA/AM-pretreated cells. These results indicate that although H(2)O(2)-induced HOCD is not directly mediated by an increase of intracellular calcium concentration, normal resting levels of intracellular calcium are required for the maintenance of MAR-associated endonuclease in an active form. PMID- 11434976 TI - Alternate cadmium exposure differentially affects amino acid metabolism within the hypothalamus, median eminence, striatum and prefrontal cortex of male rats. AB - This work was designed to analyze the possible changes in glutamate, aspartate and glutamine content induced by cadmium exposure in the hypothalamus, striatum and prefrontal cortex of rats, using an alternate schedule of metal administration. Pubertal-adult differences were also evaluated. In adult control rats, glutamate and aspartate contents in the anterior hypothalamus decreased as compared to pubertal controls. After cadmium administration from day 30 to 60 of life, the content of anterior hypothalamic glutamate and aspartate diminished. In adult control animals, the glutamine content increased in mediobasal hypothalamus as compared to pubertal controls. After cadmium exposure from day 30 to 60 of life, the mediobasal glutamine content increased, and after cadmium treatment from day 60 to 90 of life, the mediobasal aspartate content decreased. In adult control rats the content of glutamine, glutamate and aspartate of the posterior hypothalamus decreased significantly. After cadmium administration in pubertal animals, posterior hypothalamic contents of glutamine, glutamate and aspartate diminished. Cadmium treatment of adult animals caused a decrease in glutamine content, as compared to controls. In adult control rats, only glutamate and aspartate content increased in the prefrontal cortex as compared to the values found in pubertal controls. When cadmium was administered to adult animals, only the aspartate content decreased. In the striatum, cadmium decreased the glutamine and aspartate contents when administered from day 60 to 90 of life. These data suggest that cadmium differentially affects amino acid metabolism in the hypothalamus, striatum and prefrontal cortex. Age-dependent effects of cadmium on these brain areas appeared to have occurred. PMID- 11434977 TI - Dehydroepiandrosterone sulfate (DHEAS) suppresses P2X purinoceptor-coupled responses in PC12 cells. AB - Some steroids rapidly alter neuronal excitability through interaction with neurotransmitter-gated ion channels in addition to their well-known genomic effects via intracellular steroid receptors. Such effects were found in GABA receptor, nicotinic receptors, yet not investigated in P2X purinoceptors. In this study, the effects of dehydroepiandrosterone sulfate on the P2 purinoceptor was investigated. Results show that dehydroepiandrosterone sulfate acutely inhibits P2X purinoceptor functions in PC12 cells. Dehydroepiandrosterone sulfate suppressed ATP-induced cytosolic free calcium concentration ([Ca(2+)](i)) rise, cytosolic free sodium concentration ([Na(+)](i)) rise, and dopamine secretion in the presence of external calcium, but had no effect on ATP-induced [Ca(2+)](i) rise in the absence of external calcium or on UTP-induced [Ca(2+)](i) rise in the absence or presence of external calcium. Our data show that dehydroepiandrosterone sulfate exerted its effect on P2X, but not on the P2Y purinoceptors found in PC12 cells. Estradiol and estrone have similar effects on P2X purinoceptor, but dehydroepiandrosterone and progesterone do not. PMID- 11434978 TI - In vivo characterization of basal amino acid levels in subregions of the rat nucleus accumbens: effect of a dopamine D(3)/D(2) agonist. AB - Recent evidence demonstrates that two subdivisions of the nucleus accumbens, the dorsolateral core and the ventromedial shell can be distinguished by morphological, immunohistochemical and chemoarchitectural differences. In the present study, we measured basal levels of amino acids in microdialysates from both the shell and core subterritories of the nucleus accumbens in freely moving rats using HPLC with fluorescence detection. The effect of the dopamine D(3)/D(2) receptor agonist quinelorane (30 microg/kg s.c.) was then investigated in both subregions. With the exception of glutamate, histidine, and serine, which showed similar levels in both subterritories, alanine, arginine, aspartate, gamma aminobutyric acid, glutamine, and tyrosine were significantly higher in the shell compared with the core. In contrast, taurine levels were significantly lower in the shell than in the core. A particularly striking difference across subregions of the nucleus accumbens was observed for basal GABA levels with a shell/core ratio of 18.5. Among all the amino acids investigated in the present study, quinelorane selectively decreased dialysate GABA levels in the core subregion of the nucleus accumbens. The results of the present study point to specific profiles of both shell and core in terms of: (1) basal chemical neuroanatomical markers for amino acids; and (2) GABAergic response to the DA D(3)/D(2) agonist quinelorane. PMID- 11434979 TI - Acetyl- and butyrylcholinesterase molecular forms in normal and streptozotocin diabetic rat retinal pigment epithelium. AB - We studied the composition of molecular forms of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in normal and streptozotocin-induced diabetic rat retinal pigment epithelium (RPE). Tissues were sequentially extracted with saline (S(1)) and saline-detergent buffers (S(2)). About a 50% decrease in AChE molecular forms was observed in the diabetic RPE compared to the controls. Approximately 70% of the BChE activity in normal RPE was brought into solution and evenly distributed in S(1) and S(2). Analysis of the fractions from RPE revealed the presence of G(A)(1), G(A)(4) and a small proportion of G(H)(4) BChE forms in S(1); whereas G(A)(4) and G(A)(1) molecules predominate in S(2). A 40% decrease in the activity of G(A)(4) in S(2) was observed in the diabetic RPE. Our results show that diabetes caused a remarkable decrease in the activity of cholinesterases molecular forms in the RPE. This might be related to the alterations observed in diabetic retinopathy. PMID- 11434980 TI - Synaptosomal and vesicular accumulation of L-glutamate, L-aspartate and D aspartate. AB - We examined the vesicular accumulation of the excitatory amino-acid (EAA) neurotransmitters, L-glutamate and L-aspartate, together with the non metabolisable EAA analogue D-aspartate. Synaptosomes derived from whole brain were incubated in various concentrations of [3H]-amino acids under conditions to facilitate vesicular turnover. Synaptosomes were then lysed in hypotonic medium and vesicles immunoprecipitated with monoclonal anti-synaptophysin antibodies coupled to sepharose beads. Using this method, saturable vesicular accumulation was observed for [3H]-L-glutamate, [3H]-L-aspartate, and [3H]-D-aspartate but not for the excitatory amino acid receptor ligands [3H]-AMPA or [3H]-kainate. Vesicular accumulation (t(1/2)=7.45 min) was markedly slower than synaptosomal accumulation (t(1/2)=1.03 min) and was substantially reduced at 4 degrees C. Maximal accumulation of [3H]-L-glutamate, [3H]-L-aspartate, and [3H]-D-aspartate was estimated to be 98, 68, and 112 pmol/mg of synaptosomal protein, respectively, and uptake affinities 1.6, 3.4, and 2.1 mM, respectively. Maximal accumulation of [3H]-L-glutamate was non-competitively inhibited by both 100 microM unlabeled L-aspartate and 100 microM D-aspartate, suggesting that all are accumulated into a common vesicular pool by different transporters. PMID- 11434981 TI - Signaling and gene expression in the neuron-glia unit during brain function and dysfunction: Holger Hyden in memoriam. AB - Holger Hyden demonstrated almost 40 years ago that learning changes the base composition of nuclear RNA, i.e. induces an alteration in gene expression. An equally revolutionary observation at that time was that a base change occurred in both neurons and glia. From these findings, Holger Hyden concluded that establishment of memory is correlated with protein synthesis, and he demonstrated de novo synthesis of several high-molecular protein species after learning. Moreover, the protein, S-100, which is mainly found in glial cells, was increased during learning, and antibodies towards this protein inhibited memory consolidation. S-100 belongs to a family of Ca(2+)-binding proteins, and Holger Hyden at an early point realized the huge importance of Ca(2+) in brain function. He established that glial cells show more marked and earlier changes in RNA composition in Parkinson's disease than neurons. Holger Hyden also had the vision and courage to suggest that "mental diseases could as well be thought to depend upon a disturbance of processes in glia cells as in the nerve cells", and he showed that antidepressant drugs cause profound changes in glial RNA. The importance of Holger Hyden's findings and visions can only now be fully appreciated. His visionary concepts of the involvement of glia in neurological and mental illness, of learning being associated with changes in gene expression, and of the functional importance of Ca(2+)-binding proteins and Ca(2+) are presently being confirmed and expanded by others. This review briefly summarizes highlights of Holger Hyden's work in these areas, followed by a discussion of recent research, confirming his findings and expanding his visions. This includes strong evidence that glial dysfunction is involved in the development of Parkinson's disease, that drugs effective in mood disorders alter gene expression and exert profound effects on astrocytes, and that neuronal-astrocytic interactions in glutamate signaling, NO synthesis, Ca(2+) signaling, beta adrenergic activity, second messenger production, protein kinase activities, and transcription factor phosphorylation control the highly programmed events that carry the memory trace through the initial, signal-mediated short-term and intermediate memory stages to protein synthesis-dependent long-term memory. PMID- 11434982 TI - Methyl-4-phenylpyridinium (MPP(+))-evoked dopamine release from rat striatal slices: possible roles of voltage-dependent calcium channels and reverse dopamine transport. AB - We examined the properties of voltage-dependent Ca(2+) channels (VDCCs) mediating 1-methyl-4-phenylpyridinium (MPP(+))-evoked [3H]DA release from rat striatal slices. In some cases, the Ca(2+)-independent efflux of neurotransmitters is mediated by the high-affinity neurotransmitter-uptake systems. To determine whether such a mechanism might be involved in MPP(+)-evoked [3H]DA release. MPP(+) (1,10 and 100 microM) evoked the release of [3H]DA from rat striatal slices in a concentration-dependent manner. In the absence of Ca(2+), MPP(+) (10 and 100 microM)-evoked [3H]DA release was significantly decreased to approximately 50% of control (a physiological concentration of Ca(2+)). In the presence of Ca(2+), nomifensine (0.1,1 and 10 microM) dose-dependently and significantly inhibited the MPP(+)-evoked release of [3H]DA. Nomifensine (1 and 10 microM) also dose-dependently and significantly inhibited the MPP(+)-evoked release of [3H]DA under Ca(2+)-free conditions. MPP(+)-evoked [3H]DA release was partly inhibited by nicardipine (1 and 10 microM), an L-type Ca(2+) channel blocker. On the other hand, the N-type Ca(2+) channel blocker omega-conotoxin GVIA (omega-CTx-GVIA) (1 and 3 microM) did not affect this release. omega agatoxin-IVA (omega-Aga-IVA) at low concentrations (0.1 microM), which are sufficient to block P-type Ca(2+) channels alone, also had no effect. On the other hand, MPP(+)-evoked [3H]DA release was significantly decreased by high concentrations of omega-Aga-IVA (0.3 microM) that would inhibit Q-type Ca(2+) channels. In addition, application of the Q-type Ca(2+) channel blocker omega conotoxin-MVIIC (omega-CTx-MVIIC) (0.3 and 1 microM) also significantly inhibited MPP(+)-evoked [3H]DA release. These results suggest that MPP(+)-evoked [3H]DA release from rat striatal slices is largely mediated by Q-type Ca(2+) channels, and the Ca(2+)-independent component is mediated by reversal of the DA transport system. PMID- 11434984 TI - Toxicological review of inorganic phosphates. AB - Inorganic phosphate salts are widely used as food ingredients and in a variety of commercial applications. The United States Food and Drug Administration (FDA) considers inorganic phosphates "Generally Recognized As Safe" (GRAS) (FDA, 1973a, 1979) [FDA: Food and Drug Administration 1973a. GRAS (Generally Recognized as Safe) food ingredients-phosphates. NTIS PB-221-224, FDA, Food and Drug Administration, 1979. Phosphates; Proposed Affirmation of and Deletion From GRAS Status as Direct and Human Food Ingredients. Federal Register 44 (244). 74845 74857, 18 December (1979)] and the European Union (EU) allows inorganic phosphates to be added directly to food (EU Directive 95/2/EC as amended by 98/72/EC). In this review, data on the acute, subchronic and chronic toxicity, genotoxicity, teratogenicity and reproductive toxicity from the published literature and from unpublished studies by the manufacturers are reviewed. Based on the toxicity data and similar chemistry, the inorganic phosphates can be separated into four major classes, consisting of monovalent salts, divalent salts, ammonium salts and aluminum salts. The proposed classification scheme supports the use of toxicity data from one compound to assess the toxicity of another compound in the same class. However, in the case of eye and skin irritation, the proposed classification scheme cannot be used because a wide range of responses exists within each class. Therefore, the eye and skin hazards associated with an individual inorganic phosphate should be assessed on a chemical-by-chemical basis. A large amount of toxicity data exists for all four classes of inorganic phosphates. The large and comprehensive database allows an accurate assessment of the toxicity of each class of inorganic phosphate. Overall, all four classes of inorganic phosphates exhibit low oral, inhalation and dermal toxicities. Based on these data, humans are unlikely to experience adverse effects when the daily phosphorus consumption remains below 70 mg/kg/day (JECFA, 1964, 1982a) [JECFA (Joint FAO/WHO Expert Committee on Food Additives 1964. Specifications for the Identity and Purity of Food Additives and their Toxicological Evaluation) Emulsifiers, Stabilizers, Bleaching, and Maturing Agents. Technical Report Series of the World Health Organization 281; ECFA (Joint FAO/WHO Expert Committee on Food Additives 1982a. Phosphoric Acid and Phosphate Salts. ICS/FA/82)]. PMID- 11434985 TI - Masking of antioxidant capacity by the interaction of flavonoids with protein. AB - The effect of antioxidants is often executed in complex biological mixtures where various interactions may take place. Therefore, the antioxidant capacity of antioxidants in blood plasma is examined. The assay used is the trolox equivalent antioxidant capacity (TEAC). This method gives the antioxidant capacity of a compound by measuring spectrophotometrically the disappearance of the blue/green stable ABTS [2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)] radical, caused by scavenging. The results show that the antioxidant capacity of quercetin, rutin, catechin or 7-monohydroxyethylrutoside (monoHER) and blood plasma is not additive. This is partly due to interactions between the antioxidant and plasma proteins. However, the antioxidant capacity of alpha tocopherol, which also binds to protein, is not affected by the interaction. This means that besides the antioxidant capacity of the compound itself, the environment in which the antioxidant has to execute his function is important. PMID- 11434986 TI - Garlic-induced alteration in rat liver and kidney morphology and associated changes in endogenous antioxidant status. AB - The effects of chronic garlic intake on various endogenous antioxidant enzymes and lipid peroxidation on two major organs, the liver (L) and kidneys (K), were investigated. Wistar albino rats were fed with fresh garlic homogenate daily by gavage in three different doses (250, 500 and 1000 mg/kg/day) for 30 days. After this period, rats were sacrificed and liver and kidneys were harvested for biochemical estimation. In comparison to saline-treated rats, the 250 mg/kg/day dose significantly (P<0.02) reduced thiobarbituric acid reactive substances (TBARS) (L: 187.48+/-9.23 vs 150.66+/-11.45; K: 177.38 15.88 vs 120.66+/-9.39 nmol/g wet. weight) and glutathione peroxidase (GPx) (L: 0.2438+/-0.05 vs 0.0046+/-0.0005; K: 0.1459+/-0.034 vs 0.0055+/-0.0003 U/mg protein). There was no change in catalase and reduced glutathione (GSH) but superoxide dismutase (SOD) increased significantly (P<0.01) (L: 5.49+/-0.76 vs 18.38+/-2.26; K: 11.47+/-1.48 vs 21.22+/-3.19 U/mg protein). Both 500 and 1000 mg/kg/day doses significantly (P<0.05) reduced endogenous antioxidants (catalase and SOD) without altering TBARS. A 1000 mg/kg/day dose of garlic caused marked histopathological and ultrastructural changes in both liver and kidneys. The results suggest that garlic in low doses has the potential to enhance the endogenous antioxidant status, although at higher doses a reversal of these effects is observed. The present study also highlights the potential ability of a high dose of garlic to induce morphological changes in the liver and kidneys, indicating the need to identify a safe dose range for garlic. PMID- 11434987 TI - Oral toxicity of a tocotrienol preparation in rats. AB - Tocotrienols are added as antioxidants to food. As there have been no reports of toxicological evaluation, a 13-week oral toxicity study was performed in Fischer 344 rats of both sexes at dose levels of 0 (group 1), 0.19 (group 2), 0.75 (group 3) and 3% (group 4) of a preparation in powdered diet. Suppression of body weight gain was observed in group 4 males. On hematological examination, significant decrease in mean corpuscular volume (MCV) was observed in all treated males. Platelets were significantly reduced in group 3 and 4 males. Hemoglobin concentration, MCV, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration were significantly decreased in group 3 and 4 females and hematocrit in group 4 females. On serum biochemical examination, increase in the albumin/globulin ratio (A/G) and alkaline phosphatase in all treated males, elevated alanine transaminase in group 4 of both sexes and increases in asparagine transaminase and gamma-glutamyl transaminase in group 4 females were observed. With regard to relative organ weights, liver weights in group 4 of both sexes and adrenal weights in all treated males demonstrated an increase, and ovary and uterus weights in group 4 females were reduced. Histopathologically, slight hepatocellular hypertrophy in group 3 and 4 males, and reduction of cytoplasmic vacuolation in the adrenal cortical region in group 4 males were observed. Because of pathological changes in male liver and hematological changes in females, the no-observed-adverse-effect level (NOAEL) was concluded to be 0.19% in the diet (120 mg/kg body weight/day for male rats and 130 mg/kg body weight/day for female rats). As a decrease in MCV, an increase in the A/G, elevation of alkaline phosphatase and increase in adrenal weight were observed in all treated males, a no-observed-effect level (NOEL) could not be determined in this examination. PMID- 11434988 TI - An assessment of the carcinogenic potential of shea oleine in the rat. AB - Shea oleine, an oil fraction derived from the nut of the tree Butyrospermum parkii, is used as a frying oil. As part of a series of studies, this investigation examined the carcinogenic potential of 15% (w/w) shea oleine in comparison with 15% (w/w) sheanut oil, and palm oil following dietary administration to rats over 104 weeks. The assessment comprised an evaluation of mortality, clinical signs, body weight, food intake, clinical pathology, organ weights and macroscopic and histopathological examination plus tumour type and incidence evaluation. Results showed that shea oleine produced no adverse effects and no evidence of tumorigenic potential compared to other commercially available sheanut and palm oils in the rat. Notable differences were confined to reduced body weight gain and food intake, reduced cholesterol and increased alkaline phosphatase levels, reduced heart weight and an increased incidence of pulmonary lipidosis with shea oleine diets. The latter effect may reflect a naturally lower incidence of this finding with palm oil diets. Tumour findings, specific to shea oleine diets, were restricted to an increase in the number of hepatomas for females, pancreatic exocrine adenomas for males and skin keratoacanthomas for males fed shea oleine diets. The increase in the incidence of hepatomas with treatment was thought to be related to the high fat content of the diets. The incidence of these tumour findings was similar to that given in published data for the Wistar rat, or the 'in house' values for tumour incidence in rats fed high-fat diets. In conclusion, none of the findings in this study were considered to be adverse effects. In comparison with other commercially available edible oils, shea oleine showed no tumorigenic potential following dietary administration at 7.5 g/kg/day in the rat. PMID- 11434989 TI - The relationship among microsomal enzyme induction, liver weight and histological change in beagle dog toxicology studies. AB - The present study represents a retrospective analysis of hepatic microsomal enzyme induction data collected over a period of years for the beagle dog. Comparisons were completed for up to six enzyme activities and P450 content versus histopathological examination of the liver for hepatic changes and serum chemistry data analysis for markers indicative of hepatic injury. In addition, qualitative comparisons were made for these compounds to data reported in the rat by the same authors. In this analysis of canine study data for nine different compounds comprising five different pharmacological classes, significant elevations in several microsomal enzyme activities were observed under study conditions that did not result in liver weight increases, histological changes or serum chemistry changes that would be indicative of hepatocellular or hepatobiliary damage. Despite some species differences in cytochrome P450 homologues, for this compound set, there was clearly a general association between the response in dog liver and that of the rat liver. Compounds that elicited significant increases in more than one canine P450 endpoints were also likely to produce an inductive response in rat liver; however, the magnitude of the response and the P450 endpoint involved were not always identical. We conclude that hepatic drug-metabolizing enzyme induction in the beagle dog liver is typically a benign adaptive response, which parallels that reported previously in the rat. PMID- 11434990 TI - Deoxynivalenol-induced immunomodulation of human lymphocyte proliferation and cytokine production. AB - Mycotoxins are a structurally diverse group of secondary metabolites produced by different genera of fungi, and include deoxynivalenol (DON), T-2 toxin, aflatoxin B1 (AFB1) and fumonisin B1 (FB1). Despite widespread human exposure and potent immunomodulation in animals, their effects on the human immune system remain to be defined. In this study, the effect of these toxins on human lymphocyte proliferation was evaluated using the MTT assay. Additionally, the effect of DON on cytokine profiles was measured. A 50% inhibition in cell proliferation was observed with a DON concentration of 216 ng/ml. T-2 toxin was more potent with 50% inhibition between 1 and 5 ng/ml. Negligible effects were observed with AFB1 and FB1, and a mixture of DON with either FB1 or AFB1 did not show any synergistic effects in this assay. Short-term treatment of PHA-stimulated lymphocytes with DON (100, 200 and 400 ng/ml) modulated the kinetics of IL-2, IL 4 and IL-6 production. IL-2 levels were up to 12-fold higher (P<0.05) in comparison to control levels at toxin concentrations of 200 and 400 ng/ml 72 h after treatment. IL-4 levels were only slightly elevated and IL-6 levels were slightly inhibited by these DON concentrations. The kinetics of cytokine production was followed for an extended period of 8-9 days at DON concentrations of 200 and 400 ng/ml. At the lower DON concentration (200 ng/ml), IL-2 levels were elevated 17-25-fold with a concomitant mild elevation in IFN-gamma. Consistent with earlier experiments, IL-6 levels were slightly suppressed by DON at this concentration. At 400 ng/ml, IL-2 levels were again significantly (P<0.05) elevated until 6 days post-treatment, while the effects on IL-4 and IL-6 were less marked. These data suggest DON has potent effects on human lymphocyte cytokine production which merit investigation in exposed human populations. PMID- 11434991 TI - Lack of effect of coumarin on the formation of micronuclei in an in vivo mouse micronucleus assay. AB - Coumarin was tested for its potential to cause genotoxic effects in mouse bone marrow cells using an in vivo micronucleus assay. Male and female Swiss mice were administered a single oral dose of coumarin at 50, 100 or 200 mg/kg by gavage in corn oil vehicle. Control animals received only the vehicle. Groups of male mice were also administered mitomycin C at 0.75 mg/kg and served as positive controls. At 24 h after treatment, mice from all dose levels, and at 48 h after treatment, mice from the high dose level only were sacrificed. Bone marrow cells were collected and assayed for the presence of micronuclei. Coumarin did not cause any increase in the incidence of micronucleated polychromatic erythrocytes in male or female mice at any of the dose levels, the positive control mitomycin C produced a significant increase. There was no evidence of coumarin or mitomycin C treatment related cytotoxicity to bone marrow cells. The results of this study demonstrate that coumarin is negative in the mouse in vivo micronucleus assay. PMID- 11434992 TI - Use of the alkaline comet assay for industrial genotoxicity screening: comparative investigation with the micronucleus test. AB - We evaluated the suitability of the alkaline comet assay as a screening test in industrial routine testing of new chemicals. Thirty-six pharmaceutical compounds with unknown genotoxic potential were tested comparatively in the comet assay and micronucleus test (MNT) using V79 Chinese hamster cells. The comparison of results is generally based on at least two independent experiments, each with two replicate cultures at a minimum of three concentrations. We found a high degree of concordance between results of the comet assay and MNT. All compounds with negative MNT results were also negative in the comet assay. All positive compounds in the comet assay were also positive in the MNT. However, 16 of 38 positive MNT results were negative in the comet assay. Some of the contrary findings may be due to aneugenic effects, which are detected in the MNT but not in the comet assay. However, the majority of the contrary results may be a consequence of cytotoxicity, which can induce elevated micronucleus frequencies but may not lead to positive effects in the comet assay. Additional data of 39 compounds tested in the Ames test and the comet assay were compared. Four of these compounds that were Ames positive were also positive in the comet assay. However, the comet assay also detected 16 compounds that were negative in the Ames test. We believe that the comet assay in vitro is a useful, fast screening system in mammalian cells that can be used in a test battery during drug development. PMID- 11434993 TI - In vivo genotoxic effect of potassium dichromate in mice leukocytes using comet assay. AB - Hexavalent chromium is a well-known mutagen and carcinogen. In the present investigation, single-/double-stranded DNA breaks by potassium dichromate (K2Cr2O7) in mice, a sensitive model for genotoxic effects, have been studied in vivo using alkaline single-cell gel electrophoresis (SCGE)/comet assay. Mice were administered orally with a range of doses starting from 0.59 to 76.0 mg/kg body weight of K2Cr2O7 and samples of whole blood were collected at 24, 48, 72, 96 h, week 1 and week 2 post-treatment for alkaline SCGE assay to study DNA damage. The rationale for using leukocytes was to reflect biomarker analysis in humans. Significant increase in mean comet tail length (5.7-24.25 microM) indicating DNA damage was observed at all the doses with K2Cr2O7 when compared with controls (3.26 microM). Maximum increase in mean comet tail length was observed at 9.5 mg/kg body weight at 48 h post-treatment (24.25 microM). The mean comet tail length showed a clear dose-dependent increase from 0.59 to 9.5 mg/kg body weight and a dose-dependent decrease in higher doses (19.0-76.0 mg/kg body weight). A gradual decrease in the tail lengths from 72 h post-treatment was observed by the second week, and values had returned to control levels at all doses, indicating repair of the damaged DNA and/or loss of heavily damaged cells. The study also reveals that comet assay is a sensitive and rapid method for detecting DNA damage caused by heavy metals such as chromium (Cr). PMID- 11434994 TI - Developmental toxicity of sodium fluoride measured during multiple generations. AB - Sodium fluoride (NaF) has been used to fluoridate drinking water in the United States since the mid 1940s. Because of the lack of reliable studies on the multigeneration effects of the compound, NaF (0, 25, 100, 175 or 250 ppm in drinking water) was given to rats continuously during three generations. Parental (F0) generation rats were treated for 10 weeks and mated within groups. At gestation day 20, caesarean sections were performed and eight F0 females per group and their litters (F1) were observed for implant status, fetal weight and length, sex and morphological development. The remaining F0 females (29-32 per group) were allowed to litter. F1 offspring (36 of each sex per group) were mated within groups, and caesarean sections were performed at gestation day 20. The F1 females and their litters (F2) were observed for implant status, fetal weight and length, sex and morphological development. In addition, F2 fetuses were evaluated for internal (soft-tissue) and skeletal development. Decreased fluid consumption for F0 and F1 dams at 175 and 250 ppm was attributed to decreased palatability of the solution. No dose-related effects in feed consumption or mean body weight gain were observed in either F0 or F1 females. Numbers of corpora lutea, implants, viable fetuses and fetal morphological development were similar in all groups. No dose-related anomalies in internal organs were observed in F2 fetuses. Ossification of the hyoid bone of F2 fetuses was significantly decreased at 250 ppm. Because of the decreased ossification of the hyoid bone, 250 ppm is considered the effect level. PMID- 11434995 TI - Evaluation of subchronic toxicity of n-butyl acetate vapor. AB - The subchronic toxicity of n-butyl acetate (nBA), a common industrial solvent, was tested in rats in a 13-week inhalation study. Male and female Sprague-Dawley (SD) rats were exposed to concentrations of 0, 500, 1500 or 3000 ppm nBA for 6 h per day, 5 days per week for 13 consecutive weeks. Transient signs of sedation were observed only during exposure to the 1500 and 3000 ppm concentrations. Body weights for the 1500 and 3000 ppm groups were significantly reduced. Feed consumption values for the 1500 and 3000 ppm groups were significantly lower than the control group. Weights of the liver, kidneys and spleen were significantly lower for the 3000 ppm male group; testes and adrenal gland weights for the 1500 and 3000 ppm groups and the lung weight for the 3000 ppm male group were significantly higher than for the control group. Signs of irritation of the glandular stomach and necrosis in the non-glandular stomach were observed in 3000 ppm female rats. Degeneration of the olfactory epithelium along the dorsal medial meatus and ethmoturbinates of the nasal passages of some 1500 and all 3000 ppm rats was also seen. The severity was mild to moderate for the 3000 ppm group and minimal to mild for the 1500 ppm group. No effects were observed in the lungs of any group. The no-observed-effect level (NOEL) for this study is considered to be 500 ppm. The data presented here are relevant to the toxicity risk assessment of n-butanol due to the rapid hydrolysis of nBA in vivo. PMID- 11434996 TI - Changes in concentration of pesticide residues in potatoes during washing and home preparation. AB - Monitoring of pesticide residues in potato tubers and their prepared products ("pommes frites" and chips) was undertaken. Experiments were carried out to determine changes in concentration due to the washing, peeling and cooking process (blanching and frying) to assess the stability of pesticides in potatoes and their products. Pesticide residues were quantified by using gas chromatography. Results show that malathion, HCB, lindane and p,p-DDD were predominant in potatoes and their products. The highest mean was detected in potatoes, followed by pommes frites, while the lowest mean was recorded in chips. On the other hand, potato skin samples were found to contain the highest levels of DDT and its derivatives, lindane and HCB. Peeling was necessary to remove the greatest amount of pesticides in the skin. Washing with water and/or other solutions as well as the cooking process (blanching and frying) helped to eliminate most of the pesticide residues from the potato tubers. PMID- 11434998 TI - Specialist training in OB-GYN; the future. PMID- 11434999 TI - Ovarian tissue cryopreservation and gynecologic oncology: a review. AB - Sperm cryopreservation permits young men, undergoing cancer treatments, to preserve their fertility. Ovarian tissue cryopreservation have the same goal for young women and could also be an option for children. However, only primordial follicles survive after freezing and a follicular maturation is needed after thawing. This maturation has not yet been realized in humans, pregnancies have only been obtained in animal models. As cryopreservation is yet effective in humans, many teams have already cryopreserved the ovarian tissue of patients who have nothing to lose as their follicular reserve would have been destroyed or severely depleted by cancer treatment. The preservation of fertility is rarely an issue in gynecologic oncology because it usually concerns post-menopausal women. However, they are early-onset forms of gynecologic cancers and in these cases fertility is often threatened. Ovarian tissue cryopreservation may be performed when curative or prophylactic ovariectomy must be undergone, when chemotherapy with high-dose alkylating agents is planned or when pelvic radiation is needed (particularly in cases requiring chemotherapy combined with radiotherapy). In some of these situations it would be dangerous to graft back the tissue to the patient as cancer cells could remain within the grafts, the best solution in this case would be the in vitro follicular maturation. PMID- 11435000 TI - Obstetricians say yes to maternal request for elective caesarean section: a survey of current opinion. AB - AIM: To determine what proportion of obstetricians would agree to elective pre labour CS for 'maternal request'. METHODS: Every fifth consultant on an alphabetical list of obstetricians in England and Wales obtained from the RCOG (243) was surveyed by post and asked (a) "Would you agree to perform an elective CS on a woman with an uncomplicated singleton cephalic pregnancy at term for 'maternal request?"' and (b) if yes, in relation to this 'Has your practice changed over recent years?" RESULTS: 155 questionnaires were returned (63% response rate -- four unanswered, leaving 151 for analysis). One hundred and four (69%) consultants said they would agree to 'maternal request' for CS. Of the 'yes' respondents, 62 (60%) claimed their practice had changed recently. CONCLUSION: This survey demonstrates that a majority of obstetricians are now prepared to agree to maternal request for CS in the absence of obstetric indications. PMID- 11435001 TI - A national confidential survey of obstetrician's personal preferences regarding mode of delivery. AB - OBJECTIVE: To survey the personal preferences of obstetricians regarding mode of delivery, and relate these to hospital caesarean section rates. STUDY DESIGN: A confidential, questionnaire based survey to all obstetricians working in the Republic of Ireland (n=234). RESULTS: The response rate was 71% (n=165). Seven percent of Irish obstetricians would choose an elective caesarean section for themselves (or their partners) if they were primigravida with an uncomplicated, singleton cephalic presentation at term in the absence of any clinical indication. Caesarean section was the preferred mode of delivery for 38% of respondents if the estimated foetal weight was 4.5kg. There was a highly significant association between consultant obstetricians' personal preferences of towards caesarean section and their working in a hospital with a caesarean section rate greater than 16% (P<0.005). CONCLUSIONS: Irish obstetricians' personal preferences towards elective caesarean section for an uncomplicated, cephalic pregnancy at term are significantly lower than published data examining London based obstetricians' choices. There is a consistent trend against vaginal delivery if the obstetrician is female or younger. The association between a personal preference of the consultant for abdominal delivery and the caesarean section rate of the hospital that they work in may hamper efforts to decrease the rising numbers of caesarean sections. PMID- 11435002 TI - Caesarean section by choice could fulfil the inverse care law. AB - We have assessed knowledge about caesarean sections in a group of North Staffordshire Hospital professionals and patients. Those who were more interested in having a caesarean section without medical indication were the nulliparous health professionals, the majority of whom thought they should be able to choose this option. Paradoxically, this was also the group who were most likely to consider that caesarean section is more expensive and dangerous. The explanation may be that they also felt it to be an easier, less painful and more convenient option. PMID- 11435003 TI - A survey of trainee obstetricians preferences for childbirth. AB - OBJECTIVE: To determine trainee obstetricians personal preferences regarding mode and place of delivery given various scenarios. STUDY DESIGN: An anonymous nationwide postal survey of 365 specialist registrars. RESULTS: The response rate was 76%. About 2.5% preferred a home birth. And 16% of men and 15% of women opted for elective cesarean section (CS). When faced with a proposed trial of instrumental delivery in theatre, 60% accepted and a further 12% accepted only if they could choose the obstetrician performing the delivery. Regarding a breech presentation at term, 78% would accept external cephalic version (ECV). CONCLUSIONS: The percentage of obstetricians who preferred vaginal delivery and ECV were considerably higher than previously reported, and there were no significant gender differences. This study shows a more balanced attitude from obstetricians and refutes the previously held view that they necessarily advocate high levels of intervention for themselves. PMID- 11435004 TI - False aneurysm of the uterine pedicle: an uncommon cause of post-partum haemorrhage after caesarean section treated with selective arterial embolization. AB - We report three cases of post-partum haemorrhage following caesarean delivery attributed to a false aneurysm of the uterine pedicle and treated with artery embolization. These lesion were probably post-traumatic in origin related to hysterotomy. Angiographic study of the anterior division of hypogastric arteries confirmed the diagnosis and embolization of the false aneurysm was successful in controlling the haemorrhage. PMID- 11435005 TI - The effects of mifepristone on uterine sensitivity to oxytocin and on fetal heart rate patterns. AB - OBJECTIVE: To compare the incidences of tachysystole, hypertonia and fetal heart rate (FHR) abnormalities in women treated by mifepristone plus prostaglandins (PGs), mifepristone alone or PGs alone for an unfavourable cervix. STUDY DESIGN: In this retrospective study, all women between 37 and 42 weeks were eligible for the study if they had undergone cervical ripening prior to labour induction. In group 1, the women were treated with mifepristone plus PGs (n=103). Group 2 women were treated with mifepristone alone (n=96) and group 3 women with PGs alone (n=100). Incidences of tachysystole, hypertonia and FHR abnormalities were compared. RESULTS: During induction of labour, tachysystole and hypertonia occurred more frequently in women treated with mifepristone. Severe bradycardia and recurrent late decelerations were more frequent after the initiation of oxytocin in groups 1 and 2 than in group 3. CONCLUSIONS: In this study, mifepristone increased the incidences of tachysystole, hypertonia and FHR abnormality. PMID- 11435006 TI - Failure of cervical fibronectin to predict premature delivery in a population of monofetal pregnancies with idiopathic preterm labor. AB - OBJECTIVE: The purpose of the study was to evaluate the correlation between the presence of cervical fibronectin in a high-risk population of women with symptoms of preterm labor and the occurrence of preterm delivery or the need for aggressive tocolysis. STUDY DESIGN: One hundred and thirty women presenting with symptoms of threatened preterm labor were included. Cervical sampling for detection of fibronectin was performed on admission and every day until discharge or delivery. Time to delivery, length of hospital stay, use of indomethacin, delivery before 37 weeks of GA, mean term of delivery and failure of tocolysis to prevent delivery were compared to fibronectin test results. Data were analyzed using Student's t-test for continuous variables and the chi(2) test or Fisher exact test for discrete variables. RESULTS: No correlation could be found between the results of fibronectin cervical sampling on admission and any of the outcome parameters studied. Test performances were low (sensitivity 28%, specificity 57%, positive predictive value 19%, negative predictive value 69%). Results were not modified when the findings of repeated tests were taken into account. CONCLUSION: Cervical fibronectin failed to discriminate a subgroup of symptomatic women delivering prematurely. The prognostic value of fibronectin testing was not better than clinical data in our series. This observation is in disagreement with previous studies on the diagnostic value of vaginal or cervical fibronectin in preterm labor. PMID- 11435007 TI - Is cellular fibronectin a biological marker for pre-eclampsia? AB - Cellular fibronectin (cFn) appears to be an important factor in the regulation of cell-cell interactions. It is mostly synthesised by endothelial cells and its increased circulating values are thought to correspond to the endothelial damage. As pre-eclampsia is considered a pathology of endothelial cells, our study was intended to see if this circulating protein is increased during pre-eclampsia and to determine from which moment in pregnancy these changes occur. We studied cFn longitudinally in 198 consecutive pregnant patients, with or without pathologies associated to pregnancy. We measured the cFn concentrations by ELISA four times during pregnancy with the monoclonal antibody A 134. The values of cFn increased throughout the normal pregnancy, but a significant increase was observed only after the 36th week of pregnancy (P<0.0001). The values of doubling time of cFn gradually decreased, being inverse to cFn concentrations. The values of cFn were significantly higher (3.53 microg/ml; P=0.0009) in the third trimester in the group of women who developed pre-eclampsia, as compared to normals (2.23 microg/ml). cFn could not be used as a predictor of pre-eclampsia, because the clinical symptoms of this pathology were already present at the time of measurement. Our data represent the first longitudinal study showing the increase of cFn values throughout normal and pre-eclamptic pregnancies. PMID- 11435008 TI - Human immunodeficiency virus infection in pregnant women, transmission and zidovudine therapy. AB - OBJECTIVE: To determine the rate of mother to child transmission of HIV infection in women treated with zidovudine (ZDV). PATIENTS AND METHODS: A consecutive series of 52 HIV-positive pregnant women who either received antenatal care or delivered at our institution. Forty-three women were known to be HIV positive before delivery. Forty were treated with ZDV, 36 intravenously. Twelve had no antenatal care and one refused antenatal treatment. All neonates were treated with ZDV. RESULTS: The HIV transmission rate in children born to mothers treated with ZDV was 6.6%. This compares with a transmission rate of 14% before we used ZDV. All three mothers who transmitted infection had low compliance, initiated therapy late in pregnancy or had advanced disease. CONCLUSION: Our experience supports the idea that ZDV treatment has reduced HIV transmission rate. PMID- 11435009 TI - The effect of betamethasone versus dexamethasone on fetal biophysical parameters. AB - BACKGROUND: Clinical observations suggest that betamethasone reduces maternal perception of fetal movements and short term variability, but that this dose not occur after treatment with dexamethasone. OBJECTIVES: To compare the effect of betamethasone and dexamethasone on fetal biophysical parameters. METHODS: In a randomized, prospective, double blind study, 20 courses of betamethasone and 20 courses of dexamethasone were given in random sequence to patients with imminent preterm labor. During the first 32h after initiation of treatment, fetal movements were counted by the mothers and recorded by ultrasound, and a nonstress test was performed. RESULTS: Betamethasone induced a significant decrease in fetal movements as perceived by the mother and observed by ultrasound. Fetal breathing movements also decreased. Dexamethasone did not change fetal body movements. Neither drug changed the short term variability. CONCLUSIONS: Unlike betamethasone, dexamethasone does not induce a decrease in fetal movements. Dexamethasone might, therefore, be preferred for enhancement of lung maturation in imminent preterm labor. PMID- 11435010 TI - Significance of fetal fibronectin and cytokine measurement in the cervicovaginal secretions of women at term in predicting term labor and post-term pregnancy. AB - OBJECTIVE: To determine whether fetal fibronectin (FFN) or cytokine concentrations in cervicovaginal secretions can be used to predict term labor and post-term pregnancy. STUDY DESIGN: FFN and cytokines were assayed in cervicovaginal mucus from 122 pregnant women at 29-35 weeks and weekly from week 36 to parturition. RESULTS: FFN concentrations were elevated from about 3 weeks before parturition; a correlation was found between FFN levels and sampling-to delivery intervals. Parturition was best predicted within 7 days of sampling when the FFN value was >or=50ng/ml between 36 and 41 gestational weeks. Interleukin 1beta (IL-1beta) concentrations were elevated from 3 to 4 weeks before parturition; a correlation was found between IL-1beta levels and sampling-to delivery intervals. Parturition was best predicted within 7 days of sampling, with an IL-1beta cut-off value of >or=100pg/ml. CONCLUSION: Term labor and post term pregnancy can be predicted within 7 days of sampling, using FFN and IL-1beta concentrations in cervicovaginal secretions of pregnant women. PMID- 11435011 TI - Quantitative (stereological) study of placental structures in women with pregnancy iron-deficiency anemia. AB - OBJECTIVE: To determine the compensatory morphometric changes of placental structures in cases of maternal iron-deficiency anemia. METHODS: Placentas were obtained from a random sample of 26 full-term pregnant women, and methacrylate embedded vertical sections were utilized to estimate parameters of placental structures with stereological methods. Materials were divided into three groups based on maternal hemoglobin levels: anemic (hemoglobin<100g/l), intermediate and non-anemic (hemoglobin>or=110g/l). RESULTS: Placental volume and absolute volume of the intervillous space or the chorionic villi per placenta significantly increased in the anemic group. There was a significant negative correlation between maternal hemoglobin level and absolute volume or surface area of the intervillous space or the villi. No significant difference, however, was found between the groups in the volume fraction or surface density of placental structures. CONCLUSION: Placental hypertrophy was associated with mild and moderate degree of maternal iron-deficiency anemia and the enlargement of placenta appeared to be a uniform (proportional) physiological compensatory growth. PMID- 11435012 TI - Plasma progesterone kinetics following surgical treatment of ectopic pregnancies. AB - OBJECTIVES: Comparative study of plasma progesterone and betaHCG kinetics following surgical treatment of ectopic pregnancy. STUDY DESIGN: Prospective study involving 62 patients with tubal ectopic pregnancies. Study of the kinetics of plasma progesterone and betaHCG, and the correlation coefficient between plasma progesterone and betaHCG levels during post-operative follow-up. RESULTS: Thirty-nine patients were treated by salpingostomy and 23 by salpingectomy. Analyzing the betaHCG kinetics according to treatment revealed that both curves were convergent on day 2. Progesterone kinetics differed greatly in that they appeared "parallel and confused". Analyzing the correlation between betaHCG and progesterone levels proved the absence of a significant link. CONCLUSIONS: Studying the kinetics of plasma progesterone after surgical treatment of ectopic pregnancies revealed a fast decrease in progesterone. Statistical analysis of the progesterone concentration showed that post-operative kinetics is fully independent from that of betaHCG. Progesterone therefore cannot be substituted to betaHCG for post-operative follow-up. PMID- 11435013 TI - Operative injuries during vaginal hysterectomy. AB - OBJECTIVE: To evaluate incidence, characteristics and consequences of urinary and intestinal tract injuries during vaginal hysterectomy for benign conditions. STUDY DESIGN: From January 1970 to December 1996, 3076 vaginal hysterectomies with or without additional procedures, were performed for benign conditions in our department. We retrospectively analyzed operative injury cases. RESULTS: Incidence of urinary and intestinal tract injuries were 1.7 and 0.5%, respectively. Concerning urinary tract injuries, we observed only one ureteral lesion, all others being bladder lacerations (54 cases). The bladder lacerations occured during the hysterectomy step of the surgery in 61% of cases and during the additional procedures in 39%. All bladder injuries were recognized and treated during the primary operation. We observed four cases of vesico-vaginal fistula as a consequence of these injuries; all fistulas occured after bladder laceration during the hysterectomy step of the surgical procedure. Intestinal tract injuries (16 cases) were rectal lacerations occuring during the hysterectomy step of the surgery (31% of cases) and during the additional procedures (69%). All rectal injuries were recognized and repaired during the primary operation and all healed without sequellae. CONCLUSION: Operative injuries during vaginal hysterectomy are relatively rare. They are easily recognized and treated during the primary operation without important sequellae. PMID- 11435014 TI - Changes of collagen type III in female patients with genuine stress incontinence and pelvic floor prolapse. AB - PURPOSE: Objective of this study was to determine possible changes in the quantity of type III collagen in women with genuine stress incontinence (GSI) and with pelvic relaxation or not. MATERIAL AND METHODS: Ninety-four women participated in the study and they were divided in to three groups as follow: 34 patients with GSI and pelvic relaxation (group 1), 32 patients with pelvic relaxation but without GSI (group 2) and 28 patients with neither pelvic relaxation nor GSI (group 3). All the women underwent a complete pre-operative urodynamic evaluation. The presence of collagen type III was determined by immunohistochemical technique. The X-test was used for statistical analysis. A P<0.05 was considered statistically significant. RESULTS: Collagen type III was significantly reduced (p<0.05) in patients with GSI and pelvic relaxation in both sites of biopsy (group 1), compared to patients in groups 2 and 3. Specimens from women without GSI (group 2 and group 3) had a similar density of collagen type III in both sites of biopsy. CONCLUSION: In this study, we found that women with GSI had less collagen type III around the urethra regardless of the degree of pelvic relaxation. It appears that collagen has a significant role in the maintenance of urinary continence but the mechanism by which collagen metabolism is altered remains unknown. PMID- 11435015 TI - Is platinum-based chemotherapy with paclitaxel effective in optimally debulked patients with advanced ovarian cancer? AB - OBJECTIVE: Suboptimally debulked patients with advanced ovarian cancer who are treated with a combination of cisplatin plus paclitaxel (TP therapy) have a better survival as compared to patients treated with a combination of cisplatin plus cyclophosphamide (CP therapy), but this advantage has not been demonstrated in optimally debulked patients. We performed a retrospective study to compare the effectiveness of TP therapy and CP therapy in optimally debulked patients. STUDY DESIGN: From 1991 to 1996, 87 consecutive patients with advanced ovarian cancer treated in the University Hospital Utrecht and the St. Antonius Hospital were included in the study. Overall survival (OS) of patients treated with TP or CP were compared. Multivariable Cox-regression analysis was used to calculate a hazard rate ratio (HRR) for OS. RESULTS: In the study period, 51 patients were treated with CP, and 36 patients were treated with TP. In the 18 patients with a tumorrest >2cm, there was a clear, but not statistically significant benefit from TP. In 69 patients with a tumorrest 20 novel putative ADP-ribosyltransferases, including several new potential toxins. PMID- 11435083 TI - Of staphs, streps and other things. PMID- 11435084 TI - One in the eye for river blindness. PMID- 11435085 TI - Bacterial-host-cell tethers. PMID- 11435088 TI - HCV-specific immunosuppression. PMID- 11435089 TI - Caving into caveolae. PMID- 11435104 TI - The molecular mechanisms of severe typhoid fever. AB - Salmonella typhi continues to cause severe disease in many parts of the world, its most feared complication being perforation of ulcerated Peyer's patches within the small intestine, leading to peritonitis with associated mortality. The pathogenesis of this process is not well understood. In this article, we present a theoretical mechanism as to how bacterial factors and host immunological mediators within infected tissue might contribute to the observed intestinal pathology, and propose that necrosis of the Peyer's patches observed in typhoid is caused by a mechanism similar to the Shwartzman and Koch reactions. PMID- 11435106 TI - Model systems: modeling human staphylococcal arthritis and sepsis in the mouse. AB - The staphylococci have been recognized as serious pathogens for over a century and are the etiological agent of a variety of diseases ranging from mild cutaneous infections to often fatal forms of septic arthritis, endocarditis, toxic shock syndrome and sepsis. Despite intensive efforts to halt their spread, they remain the most common cause of community- and nosocomially acquired bacteremia. Murine models of Staphylocococus aureus-mediated arthritis and sepsis exist and are being used to gain a better understanding of the host-bacterium relationship as well to develop better methods of prevention and treatment. PMID- 11435107 TI - Virulence factors of Candida albicans. AB - Candidiasis is a common infection of the skin, oral cavity and esophagus, gastrointestinal tract, vagina and vascular system of humans. Although most infections occur in patients who are immunocompromised or debilitated in some other way, the organism most often responsible for disease, Candida albicans, expresses several virulence factors that contribute to pathogenesis. These factors include host recognition biomolecules (adhesins), morphogenesis (the reversible transition between unicellular yeast cells and filamentous, growth forms), secreted aspartyl proteases and phospholipases. Additionally, 'phenotypic switching' is accompanied by changes in antigen expression, colony morphology and tissue affinities in C. albicans and several other Candida spp. Switching might provide cells with a flexibility that results in the adaptation of the organism to the hostile conditions imposed not only by the host but also by the physician treating the infection. PMID- 11435108 TI - Detecting anomalous gene clusters and pathogenicity islands in diverse bacterial genomes. AB - A gene in a genome is defined as putative alien (pA) if its codon usage difference from the average gene exceeds a high threshold and codon usage differences from ribosomal protein genes, chaperone genes and protein-synthesis processing factors are also high. pA gene clusters in bacterial genomes are relevant for detecting genomic islands (GIs), including pathogenicity islands (PAIs). Four other analyses appropriate to this task are G+C genome variation (the standard method); genomic signature divergences (dinucleotide bias); extremes of codon bias; and anomalies of amino acid usage. For example, the cagA domain of Helicobacter pylori is highly deviant in its genome signature and codon bias from the rest of the genome. Using these methods we can detect two potential PAIs in the Neisseria meningitidis genome, which contain hemagglutinin and/or hemolysin-related genes. Additionally, G+C variation and genome signature differences of the Mycobacterium tuberculosis genome indicate two pA gene clusters. PMID- 11435109 TI - The implications of a low rate of horizontal transfer in Borrelia. AB - The nature and rate of recombination can be studied by comparing the sequences of multiple genes across a set of strains. When this approach is applied to Borrelia burgdorferi, four results emerge: (1) chromosomal genes are clonal; (2) there is little or no plasmid exchange; (3) the major mode of horizontal transfer of genetic material inserts a small fragment of DNA, typically <1 kb, during recombination; and (4) the level of horizontal transfer in Borrelia is so low that there is evidence for horizontal transfer only in genes where there is positive selection for diversity, that is, positive selection for the recombinant. Thus, Borrelia can serve as a model of a low recombination taxon. The implications of these results lead us to postulate that an unknown agent that is part of the Borrelia genome mediates the horizontal transfer of small fragments of DNA; the rare transfer of small fragments of DNA excludes both DNA parasites and virulence factors from the genome. PMID- 11435110 TI - Solution structure of the LDL receptor EGF-AB pair: a paradigm for the assembly of tandem calcium binding EGF domains. AB - BACKGROUND: From the observed structure and sequence of a pair of calcium binding (cb) epidermal growth factor-like (EGF) domains from human fibrillin-1, we proposed that many tandem cbEGF domains adopt a conserved relative conformation. The low-density lipoprotein receptor (LDLR), which is functionally unrelated to fibrillin-1, contains a single pair of EGF domains that was chosen for study in the validation of this hypothesis. The LDLR is the protein that is defective in familial hypercholesterolaemia, a common genetic disorder that predisposes individuals to cardiovascular complications and premature death. RESULTS: Here, we present the solution structure of the first two EGF domains from the LDL receptor, determined using conventional NMR restraints and residual dipolar couplings. The cbEGF domains have an elongated, rod-like arrangement, as predicted. The new structure allows a detailed assessment of the consequences of mutations associated with familial hypercholesterolaemia to be made. CONCLUSIONS: The validation of the conserved arrangement of EGF domains in functionally distinct proteins has important implications for structural genomics, since multiple tandem cbEGF pairs have been identified in many essential proteins that are implicated in human disease. Our results provide the means to use homology modeling to probe structure-function relationships in this diverse family of proteins and may hold the potential for the design of novel diagnostics and therapies in the future. PMID- 11435111 TI - Thioredoxin fold as homodimerization module in the putative chaperone ERp29: NMR structures of the domains and experimental model of the 51 kDa dimer. AB - BACKGROUND: ERp29 is a ubiquitously expressed rat endoplasmic reticulum (ER) protein conserved in mammalian species. Fold predictions suggest the presence of a thioredoxin-like domain homologous to the a domain of human protein disulfide isomerase (PDI) and a helical domain similar to the C-terminal domain of P5-like PDIs. As ERp29 lacks the double-cysteine motif essential for PDI redox activity, it is suggested to play a role in protein maturation and/or secretion related to the chaperone function of PDI. ERp29 self-associates into 51 kDa dimers and also higher oligomers. RESULTS: 3D structures of the N- and C-terminal domains determined by NMR spectroscopy confirmed the thioredoxin fold for the N-terminal domain and yielded a novel all-helical fold for the C-terminal domain. Studies of the full-length protein revealed a short, flexible linker between the two domains, homodimerization by the N-terminal domain, and the presence of interaction sites for the formation of higher molecular weight oligomers. A gadolinium-based relaxation agent is shown to present a sensitive tool for the identification of macromolecular interfaces by NMR. CONCLUSIONS: ERp29 is the first eukaryotic PDI-related protein for which the structures of all domains have been determined. Furthermore, an experimental model of the full-length protein and its association states was established. It is the first example of a protein where the thioredoxin fold was found to act as a specific homodimerization module, without covalent linkages or supporting interactions by further domains. A homodimerization module similar as in ERp29 may also be present in homodimeric human PDI. PMID- 11435112 TI - Crystal structure of a novel-type archaeal rubisco with pentagonal symmetry. AB - BACKGROUND: Ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) is the key enzyme of the Calvin-Benson cycle and catalyzes the primary reaction of CO2 fixation in plants, algae, and bacteria. Rubiscos have been so far classified into two types. Type I is composed of eight large subunits (L subunits) and eight small subunits (S subunits) with tetragonal symmetry (L8S8), but type II is usually composed only of two L subunits (L2). Recently, some genuinely active Rubiscos of unknown physiological function have been reported from archaea. RESULTS: The crystal structure of Rubisco from the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 (Tk-Rubisco) was determined at 2.8 A resolution. The enzyme is composed only of L subunits and showed a novel (L2)5 decameric structure. Compared to previously known type I enzymes, each L2 dimer is inclined approximately 16 degrees to form a toroid-shaped decamer with its unique L2-L2 interfaces. Differential scanning calorimetry (DSC), circular dichroism (CD), and gel permeation chromatography (GPC) showed that Tk-Rubisco maintains its secondary structure and decameric assembly even at high temperatures. CONCLUSIONS: The present study provides the first structure of an archaeal Rubisco, an unprecedented (L2)5 decamer. Biochemical studies indicate that Tk Rubisco maintains its decameric structure at high temperatures. The structure is distinct from type I and type II Rubiscos and strongly supports that Tk-Rubisco should be classified as a novel type III Rubisco. PMID- 11435113 TI - 1.3 A structure of arylsulfatase from Pseudomonas aeruginosa establishes the catalytic mechanism of sulfate ester cleavage in the sulfatase family. AB - BACKGROUND: Sulfatases constitute a family of enzymes with a highly conserved active site region including a Calpha-formylglycine that is posttranslationally generated by the oxidation of a conserved cysteine or serine residue. The crystal structures of two human arylsulfatases, ASA and ASB, along with ASA mutants and their complexes led to different proposals for the catalytic mechanism in the hydrolysis of sulfate esters. RESULTS: The crystal structure of a bacterial sulfatase from Pseudomonas aeruginosa (PAS) has been determined at 1.3 A. Fold and active site region are strikingly similar to those of the known human sulfatases. The structure allows a precise determination of the active site region, unequivocally showing the presence of a Calpha-formylglycine hydrate as the key catalytic residue. Furthermore, the cation located in the active site is unambiguously characterized as calcium by both its B value and the geometry of its coordination sphere. The active site contains a noncovalently bonded sulfate that occupies the same position as the one in para-nitrocatecholsulfate in previously studied ASA complexes. CONCLUSIONS: The structure of PAS shows that the resting state of the key catalytic residue in sulfatases is a formylglycine hydrate. These structural data establish a mechanism for sulfate ester cleavage involving an aldehyde hydrate as the functional group that initiates the reaction through a nucleophilic attack on the sulfur atom in the substrate. The alcohol is eliminated from a reaction intermediate containing pentacoordinated sulfur. Subsequent elimination of the sulfate regenerates the aldehyde, which is again hydrated. The metal cation involved in stabilizing the charge and anchoring the substrate during catalysis is established as calcium. PMID- 11435114 TI - Crystal structure of diisopropylfluorophosphatase from Loligo vulgaris. AB - BACKGROUND: Phosphotriesterases (PTE) are enzymes capable of detoxifying organophosphate-based chemical warfare agents by hydrolysis. One subclass of these enzymes comprises the family of diisopropylfluorophosphatases (DFPases). The DFPase reported here was originally isolated from squid head ganglion of Loligo vulgaris and can be characterized as squid-type DFPase. It is capable of hydrolyzing the organophosphates diisopropylfluorophosphate, soman, sarin, tabun, and cyclosarin. RESULTS: Crystals were grown of both the native and the selenomethionine-labeled enzyme. The X-ray crystal structure of the DFPase from Loligo vulgaris has been solved by MAD phasing and refined to a crystallographic R value of 17.6% at a final resolution of 1.8 A. Using site-directed mutagenesis, we have structurally and functionally characterized essential residues in the active site of the enzyme. CONCLUSIONS: The crystal structure of the DFPase from Loligo vulgaris is the first example of a structural characterization of a squid type DFPase and the second crystal structure of a PTE determined to date. Therefore, it may serve as a structural model for squid-type DFPases in general. The overall structure of this protein represents a six-fold beta propeller with two calcium ions bound in a central water-filled tunnel. The consensus motif found in the blades of this beta propeller has not yet been observed in other beta propeller structures. Based on the results obtained from mutants of active site residues, a mechanistic model for the DFP hydrolysis has been developed. PMID- 11435115 TI - Structural characterization of the LEM motif common to three human inner nuclear membrane proteins. AB - BACKGROUND: Integral membrane proteins of the inner nuclear membrane are involved in chromatin organization and postmitotic reassembly of the nucleus. The discovery that mutations in the gene encoding emerin causes X-linked Emery Dreifuss muscular dystrophy has enhanced interest in such proteins. A common structural domain of 50 residues, called the LEM domain, has been identified in emerin MAN1, and lamina-associated polypeptide (LAP) 2. In particular, all LAP2 isoforms share an N-terminal segment composed of such a LEM domain that is connected to a highly divergent LEM-like domain by a linker that is probably unstructured. RESULTS: We have determined the three-dimensional structures of the LEM and LEM-like domains of LAP2 using nuclear magnetic resonance and molecular modeling. Both domains adopt the same fold, mainly composed of two large parallel alpha helices. CONCLUSIONS: The structural LEM motif is found in human inner nuclear membrane proteins and in protein-protein interaction domains from bacterial multienzyme complexes. This suggests that LEM and LEM-like domains are protein-protein interaction domains. A region conserved in all LEM domains, at the surface of helix 2, could mediate interaction between LEM domains and a common protein partner. PMID- 11435116 TI - The kappa-carrageenase of P. carrageenovora features a tunnel-shaped active site: a novel insight in the evolution of Clan-B glycoside hydrolases. AB - BACKGROUND: kappa-carrageenans are gel-forming, sulfated 1,3-alpha-1,4-beta galactans from the cell walls of marine red algae. The kappa-carrageenase from the marine, gram-negative bacterium Pseudoalteromonas carrageenovora degrades kappa-carrageenan both in solution and in solid state by an endoprocessive mechanism. This beta-galactanase belongs to the clan-B of glycoside hydrolases. RESULTS: The structure of P. carrageenovora kappa-carrageenase has been solved to 1.54 A resolution by the multiwavelength anomalous diffraction (MAD) method, using a seleno-methionine-substituted form of the enzyme. The enzyme folds into a curved beta sandwich, with a tunnel-like active site cavity. Another remarkable characteristic is the presence of an arginine residue at subsite -1. CONCLUSIONS: The crystal structure of P. carrageenovora kappa-carrageenase is the first three dimensional structure of a carrageenase. Its tunnel-shaped active site, the first to be reported for enzymes other than cellulases, suggests that such tunnels are associated with the degradation of solid polysaccharides. Clan-B glycoside hydrolases fall into two subgroups, one with catalytic machinery held by an ancestral beta bulge, and the other in which it is held by a regular beta strand. At subsite -1, all of these hydrolases exhibit an aromatic amino acid that interacts with the hexopyranose ring of the monosaccharide undergoing catalysis. In addition, in kappa-carrageenases, an arginine residue recognizes the sulfate ester substituents of the beta-linked kappa-carrageenan monomers. It also appears that, in addition to the nucleophile and acid/base catalysts, two other amino acids are involved with the catalytic cycle, accelerating the deglycosylation step. PMID- 11435117 TI - A structural genomics approach to the study of quorum sensing: crystal structures of three LuxS orthologs. AB - BACKGROUND: Quorum sensing is the mechanism by which bacteria control gene expression in response to cell density. Two major quorum-sensing systems have been identified, system 1 and system 2, each with a characteristic signaling molecule (autoinducer-1, or AI-1, in the case of system 1, and AI-2 in system 2). The luxS gene is required for the AI-2 system of quorum sensing. LuxS and AI-2 have been described in both Gram-negative and Gram-positive bacterial species and have been shown to be involved in the expression of virulence genes in several pathogens. RESULTS: The structure of the LuxS protein from three different bacterial species with resolutions ranging from 1.8 A to 2.4 A has been solved using an X-ray crystallographic structural genomics approach. The structure of LuxS reported here is seen to have a new alpha-beta fold. In all structures, an equivalent homodimer is observed. A metal ion identified as zinc was seen bound to a Cys-His-His triad. Methionine was found bound to the protein near the metal and at the dimer interface. CONCLUSIONS: These structures provide support for a hypothesis that explains the in vivo action of LuxS. Specifically, acting as a homodimer, the protein binds a methionine analog, S-ribosylhomocysteine (SRH). The zinc atom is in position to cleave the ribose ring in a step along the synthesis pathway of AI-2. PMID- 11435118 TI - The crystal structure of yeast thiamin pyrophosphokinase. AB - BACKGROUND: Thiamin pyrophosphokinase (TPK) catalyzes the transfer of a pyrophosphate group from ATP to vitamin B1 (thiamin) to form the coenzyme thiamin pyrophosphate (TPP). Thus, TPK is important for the formation of a coenzyme required for central metabolic functions. TPK has no sequence homologs in the PDB and functions by an unknown mechanism. The TPK structure has been determined as a significant step toward elucidating its catalytic action. RESULTS: The crystal structure of Saccharomyces cerevisiae TPK complexed with thiamin has been determined at 1.8 A resolution. TPK is a homodimer, and each subunit consists of two domains. One domain resembles a Rossman fold with four alpha helices on each side of a 6 strand parallel beta sheet. The other domain has one 4 strand and one 6 strand antiparallel beta sheet, which form a flattened sandwich structure containing a jelly-roll topology. The active site is located in a cleft at the dimer interface and is formed from residues from domains of both subunits. The TPK dimer contains two compound active sites at the subunit interface. CONCLUSIONS: The structure of TPK with one substrate bound identifies the location of the thiamin binding site and probable catalytic residues. The structure also suggests a likely binding site for ATP. These findings are further supported by TPK sequence homologies. Although possessing no significant sequence homology with other pyrophospokinases, thiamin pyrophosphokinase may operate by a mechanism of pyrophosphoryl transfer similar to those described for pyrophosphokinases functioning in nucleotide biosynthesis. PMID- 11435119 TI - Rubella. AB - Rubella, also known as German measles or three-day measles, is a self-limited, mild viral illness that poses little danger to children or adults. For the developing fetus, however, infection with rubella virus is a grave threat, capable of inducing severe anomalies and permanent disability. Despite widespread vaccination programs, populations of susceptible individuals persist, among them women of childbearing age whose pregnancies remain vulnerable to congenital rubella syndrome. In the United States, the currently used rubella vaccine employs live-attenuated virus and is contraindicated in pregnancy. Nonpregnant women receiving rubella vaccination should be advised to use effective contraception for 3 months after inoculation. These warnings persist despite the fact that no clinically significant case of congenital rubella syndrome after maternal vaccination has been reported. Obstetrician/gynecologists must be familiar with rubella and the management of pregnancies complicated by exposure to the disease. Furthermore, practitioners must actively seek and vaccinate susceptible individuals to minimize this significant threat to public health. PMID- 11435120 TI - Rubeola. AB - Rubeola, more commonly known as the "red measles" or "10-day measles," is a highly contagious exanthematous viral illness. Although most cases end with full recovery, rare but serious, even fatal sequelae such as encephalitis and pneumonia can occur. The incidence of measles worldwide has decreased dramatically since the introduction of effective vaccines. Nevertheless, outbreaks still occur among clusters of individuals, especially young adults, that vaccination programs have failed to reach. Susceptible young women are at unique risk because measles in pregnancy follows a more complicated course than in nonpregnant adults. Furthermore, pregnancies complicated by rubeola are more likely to end with spontaneous abortion or preterm delivery. In the United States, the currently used rubeola vaccine employs live-attenuated virus and is contraindicated in pregnancy. Nonpregnant women receiving rubeola vaccination should be advised to use effective contraception for 3 months after inoculation. These warnings persist despite the fact that an adverse pregnancy outcome after maternal measles vaccination has not been reported. Obstetrician/gynecologists must be familiar with rubeola and the management of pregnancies complicated by exposure to the disease. Furthermore, practitioners must actively seek and vaccinate susceptible individuals to minimize this ongoing threat to public health. PMID- 11435121 TI - Child occupant protection: a summary of current safety recommendations. AB - In 1998, 140 children younger than 1 year of age were killed in motor vehicle crashes, and nearly half of those children killed were restrained. Used correctly, child safety seats dramatically reduce a child's risk of death in a motor vehicle crash, but unfortunately, incorrect use of child seats is widespread. Obstetricians and gynecologists have a unique opportunity to counsel parents on the correct restraint for their infants and to provide recommendations for the proper restraint of older children as well. The recommendations for child occupant protection are updated with new information constantly. This article will provide an overview of the current recommendations for restraining children in motor vehicles and identify sources for up-to-date information for physicians and families. PMID- 11435122 TI - Borderline ovarian tumors. AB - Borderline ovarian tumors (BOTs) represent a small subset of epithelial ovarian cancers that were first described more than 70 years ago. Recognized in a systematic way in only the last three decades, BOTs occur most often in women in their reproductive years but are associated with an excellent long-term prognosis. The management of these tumors is primarily surgical, with the role of medical therapy a topic of ongoing research and debate. PMID- 11435123 TI - Conservative management of urinary incontinence in women. AB - Urinary incontinence (UI) is now recognized as a growing health care problem and a personal concern for women. UI is felt to be a significant aging health issue for women. Much is known about the prevalence of UI in women. Almost one third (31%) of women between the ages of 42 and 50 and two in five (38%) women over the age of 60 suffer from UI. Urine leakage varies, with about 1 in 10 women leaking enough urine that it soaks through underclothes. About one in three (30%) women have problems with incontinence during pregnancy. Society incurs a significant economic burden as a result of UI, which cost the Medicare Part A program $26.3 billion in 1995. One of the biggest obstacles to effective management of incontinence is the perception that incontinence is inevitable and irreversible, a perception almost as common among health care providers as patients. Therefore, most women do not report their UI problem to primary care practitioners. Primary care practitioners are in a key position to influence prevention, screen for incontinence, and improve outcomes of women at risk for incontinence. Current research supports the value of noninvasive, conservative treatment strategies, education, and emotional support. PMID- 11435124 TI - Interferon therapy in primary care. AB - Interferons are proteins produced by human blood cells in response to stimulation (viral infection). The natural roles of interferons are host defense and modulation of the immune system. Therapeutic uses are based on these roles. Interferon-alpha has been widely used for malignancies, skin conditions, viral infections, and myeloproliferative disorders. Interferon-beta is a standard treatment for relapsing multiple sclerosis. Interferon-gamma therapy is currently used for chronic granulomatous disease and skin lesions (human papilloma virus related and keloids), but further research is ongoing. Side effects of interferon therapy are common and limit utility. Flulike symptoms are reported by more than 75% and depression by 10-40% of interferon users. Severe adverse effects are less common but may be life threatening, including autoimmune diseases, thrombotic thrombocytopenic purpura, and acute renal failure. Limited use of interferon therapy during pregnancy has been described, with successful maternal and neonatal outcomes. Use of interferon therapy during early pregnancy is not an indication for termination. PMID- 11435125 TI - Physiological function as regulation of large transcriptional programs: the cellular response to genotoxic stress. AB - The responses to ionizing radiation and other genotoxic environmental stresses are complex and are regulated by a number of overlapping molecular pathways. One such stress signaling pathway involves p53, which regulates the expression of over 100 genes already identified. It is also becoming increasingly apparent that the pattern of stress gene expression has some cell type specificity. It may be possible to exploit these differences in stress gene responsiveness as molecular markers through the use of a combined informatics and functional genomics approach. The techniques of microarray analysis potentially offer the opportunity to monitor changes in gene expression across the entire set of expressed genes in a cell or organism. As an initial step in the development of a functional genomics approach to stress gene analysis, we have recently demonstrated the utility of cDNA microarray hybridization to measure radiation-stress gene responses and identified a number of previously unknown radiation-regulated genes. The responses of some of these genes to DNA-damaging agents vary widely in cell lines from different tissues of origin and different genetic backgrounds. While this again highlights the importance of a cellular context to genotoxic stress responses, it also raises the prospect of expression-profiling of cell lines, tissues, and tumors. Such profiles may have a predictive value if they can define regions of 'expression space' that correlate with important endpoints, such as response to cancer therapy regimens, or identification of exposures to environmental toxins. PMID- 11435126 TI - Identifying 'prime suspects': symbioses and the evolution of multicellularity. AB - The possible involvement of symbioses in the evolution of multicellularity is explored. Evidence is drawn principally from the biology of present day associations of plants and animals with prokaryotes. A particular emphasis is placed on future research opportunities in this area of biology that have been provided by the advent of specific molecular techniques and new model systems. With the application of new approaches that result from these advances, a more holistic understanding of the biology of the coevolved communities, composed of animals or plants and their associated prokaryotes, is within the reach of biologists over the next few decades. PMID- 11435127 TI - Changes in rodent-erythrocyte methemoglobin reductase system produced by two malaria parasites, viz. Plasmodium yoelii nigeriensis and Plasmodium berghei. AB - The methemoglobin reductase system plays a vital role in maintaining the equilibrium between hemoglobin and methemoglobin in blood. Exposure of red blood cells to oxidative stress (pathological/physiological) may cause impairment to this equilibrium. We studied the status of erythrocytic methemoglobin and the related reductase system during Plasmodium yoelii nigeriensis infection in mice and P. berghei infection in mastomys. Malaria infection was induced by intraperitoneal inoculation with 10(6) infected erythrocytes. The present investigation revealed a significant decrease in the activity of methemoglobin reductase, with a concomitant rise in methemoglobin content during P. yoelii nigeriensis infection in mice erythrocytes. This was accompanied with a significant increase in reduced glutathione and ascorbate levels. The activity of lactate dehydrogenase, glucose 6-phosphate dehydrogenase and glutathione reductase increased with a progressive rise in parasitemia. However, no methemoglobin or associated reductase activity was detected in normal and P. berghei-infected mastomys. P. berghei infection in mastomys resulted in an increase in the level of reduced glutathione and ascorbate in erythrocytes, and also in the activity of lactate dehydrogenase, glucose 6-phosphate dehydrogenase and glutathione reductase. These results suggest that antioxidants/antioxidant enzymes may prevent or reduce the formation of methemoglobin in the host and thereby protect the host from methemoglobinemia. PMID- 11435128 TI - Hydrocarbons of Rhodnius prolixus, a Chagas disease vector. AB - The surface hydrocarbons of the blood-sucking insect, Rhodnius prolixus, a major Chagas disease vector in Venezuela, Colombia and Central America, were characterized by capillary gas chromatography coupled to mass spectrometry (CGC MS). A total of 54 single or multicomponent peaks of saturated, straight-chain and methyl-branched hydrocarbons were identified. Major n-alkanes were n-C27, n C29, n-C31 and n-C33 hydrocarbons. In the branched fraction, methyl groups were at positions 3, 5, 7, 11, 13, 15 and 17- for monomethyl isomers, and separated by three or five methylene groups for the trimethyl or tetramethyl derivatives. For the higher molecular weight components of 37, 39 and 41 atoms in the carbon skeleton, the di-, tri- and tetramethyl branches were usually separated by three or five, and sometimes 7, 11 or 13, methylene groups. The internal hydrocarbon pool contained larger amounts of the higher molecular weight methyl-branched components. Qualitative differences among epicuticular and internal hydrocarbon compositions were detected, both in adult and nymphal stages. No significant sexual dimorphism was detected, but a significant shift in the major n-alkane components was evident from the nymphal to the adult stage, differing also in the relative amounts of the higher molecular weight methyl-branched chains. Comparison of the hydrocarbon components to that of other Chagas disease vectors is discussed. PMID- 11435129 TI - Liver gangliosides of various animals ranging from fish to mammalian species. AB - Liver gangliosides of different animal species were analyzed. Bony fish liver contained a major ganglioside that migrated faster than GM3 on thin-layer chromatography (TLC). This ganglioside was identified to be GM4 (NeuAc) by methods including product analysis after sialidase treatment and negative-ion electrospray ionization (ESI)-mass spectrometry (MS). The presence of GM4 (NeuGc) in fish liver was also demonstrated. The main ganglioside band of bovine liver consisted of two different molecular species, i.e. GD1a (NeuAc/NeuAc) and GD1a (NeuAc/NeuGc). Major gangliosides of liver tissue exhibited a distinct phylogenetic profile; GM4 was expressed mainly in lower animals such as bony fish and frog liver, whereas mammalian liver showed ganglioside patterns with smaller proportions of monosialo ganglioside species. While c-series gangliosides were consistently expressed in lower animals, they were found only in mammalian liver of particular species. No apparent trend was observed between the concentration of liver gangliosides and the phylogenetic stage of animals. The present study demonstrates the species-specific expression of liver gangliosides. PMID- 11435130 TI - Muscle glycogen, lactate and glycerol-3-phosphate concentrations of larval and young adult lampreys in response to exercise. AB - When stimulated, the ammocoetes (larvae) of Geotria australis swim continuously at a moderate rate for only approximately 20 min, whereas the downstream migrants (young adults) of this species did not become exhausted following similar swimming activity over the same period. Mean concentrations of muscle glycogen in ammocoetes declined during exercise, but returned to resting levels within 30 min of recovery, whereas those in young adults changed little during the corresponding periods. Moreover, muscle lactate concentrations of ammocoetes rose markedly during exercise and the first 30 min of recovery, before declining significantly, while those of young adults remained similar during and immediately after exercise. Calculations, using the glycogen and lactate concentrations immediately after exercise, suggest that during exercise glycogen is, to some extent, utilised anaerobically (approx. 24%) by ammocoetes, but only aerobically by young adults. Furthermore, since young adults used only a small amount of glycogen, they presumably metabolised triacylglycerol aerobically to produce energy. Muscle glycerol-3-phosphate levels were far higher prior to and immediately after exercise in downstream migrants than in ammocoetes and then declined precipitously. The above trends in muscle glycogen and lactate of larval G. australis parallels, to some degree, those recorded by other workers for upstream migrant Petromyzon marinus that had been exercised to exhaustion. PMID- 11435131 TI - Molecular cloning and expression of connexin 32.3 cDNA in the ovary from the red seabream (Pagrus major). AB - This study examined connexin (Cx) gene activity in relation to oocyte maturation in the red seabream (Pagrus major) ovary. Mixed primers for the polymerase chain reaction (PCR) were designed based on the high sequence homology of selected regions of known Cx genes. PCR-amplified cDNA fragments generated by 3' and 5' rapid amplication of cDNA ends (RACE) were combined to generate full-length cDNA sequences. The 1212-bp cDNA has an open reading frame encoding 282 amino acids, with a molecular mass of 32.3 kDa (red seabream Cx32.3). Hydropathy plots of red seabream Cx32.3 show the four typical major hydrophobic and four major hydrophilic regions of Cx proteins. Typical Cx consensus sequences are observed in the first and second extracellular loops. The ovarian follicles of matured female seabream were incubated in the presence of 17alpha,20beta-dihydroxy-4 pregnen-3-one (DHP, 10 ng/ml), gonadotropin (GtH)-I (300 ng/ml) and GtH-II (300 ng/ml). Northern blot analysis of poly(A)(+) RNA extracted from the ovarian follicles were hybridized with red seabream Cx32.3 and beta-actin probes. The transcription level of PmCx32.3 in the presence of DHP, PmGtH-I and PmGtH-II was significantly higher than in the control. PMID- 11435132 TI - Proteins immunoreactive with antibody against a human leptin fragment are found in serum and tissues of the sea lamprey, Petromyzon marinus L. AB - An affinity-purified, polyclonal antibody raised against a peptide corresponding to amino acids 137-156 at the carboxy terminus of human leptin (16 kD) was used to search for immunoreactive protein(s) in the lamprey, Petromyzon marinus. Immunoblots of serum from different phases of the life cycle showed the presence of a 65-kD immunoreactive protein in the larvae and all stages of metamorphosis but not in feeding juvenile and upstream migrant adults. Extracts of tissues known to store fat were also examined using the same antibody. Muscle and fat column from all phases tested (larvae, stage 2 and 4 metamorphosing animals, feeding juveniles and upstream migrants) showed 100- and 50-kD immunoreactive proteins. Extracts of nephric fold, the primary site of fat storage during metamorphosis, lacked the 100-kD protein but had the 50 kD; they also had a 16 kD immunoreactive protein not found in the other tissues. The immunoreactivity of the proteins of both serum and tissue extracts was blocked by pretreatment of the antibody with the leptin-derived antigen. The results indicate that P. marinus has proteins that share at least one epitope with mammalian leptin. PMID- 11435133 TI - Amino acid sequence of squid troponin C. AB - The complete amino acid sequence of squid Todarodes pacificus troponin C (TnC), which was shown to bind only 1 mol Ca(2+)/mol, was determined by both the Edman and cDNA methods. The squid TnC is composed of 147 amino acids including an unblocked Pro at the N-terminus and the calculated molecular weight is 17003.9. Among the four potential Ca(2+)-binding sites, namely sites I-IV from the N terminus, only site IV completely satisfied the consensus amino acid sequence for the active Ca(2+)-binding loop. This indicates that squid TnC possesses a single Ca(2+)-binding site at the site IV as scallop TnCs [Nishita et al., J. Biol. Chem. 269 (1994) 3464-3468; Ojima et al., Arch. Biochem. Biophys. 311 (1994) 272 276). The sequence homology of squid TnC to TnCs of scallop, arthropods, and rabbit was 61%, 31-38%, and 31%, respectively. In the sequence of the central D/E helix region of squid and scallop TnCs, a deletion of three amino acids was required to maximize the homology with the other TnCs. PMID- 11435134 TI - Biochemical properties of porcine white adipose tissue mitochondria and relevance to fatty acid oxidation. AB - The capacity of white adipose tissue mitochondria to support a high beta oxidative flux was investigated by comparison to liver mitochondria. Based on marker enzyme activities and electron microscopy, the relative purity of the isolated mitochondria was similar thus allowing a direct comparison on a protein basis. The results confirm the comparable capacity of adipose tissue and liver mitochondria for palmitoyl-carnitine oxidation. Relative to liver, both citrate synthase and alpha-ketoglutarate dehydrogenase were increased 7.87- and 10.38 fold, respectively. In contrast, adipose tissue NAD-isocitrate dehydrogenase was decreased (2.85-fold). Such modifications in the citric acid cycle are expected to severely restrict citrate oxidation in porcine adipose tissue. Except for cytochrome c oxidase, activities of the enzyme complexes comprising the electron transport chain were not significantly different. The decrease in adipose cytochrome c oxidase activity could partly be attributed to a decreased inner membrane as suggested by lipid and enzyme analysis. In addition, Western blotting indicated that adipose and liver mitochondria possess similar quantities of cytochrome c oxidase protein. Taken together these results indicate that not only is the white adipose tissue protoplasm relatively rich in mitochondria, but that these mitochondria contain comparable enzymatic machinery to support a relatively high beta-oxidative rate. PMID- 11435135 TI - Purification and partial characterisation of alpha(2)-antiplasmin and plasmin(ogen) from ostrich plasma. AB - This study reports the isolation and partial characterisation of the ostrich serpin, alpha(2)AP, and its target enzyme, ostrich plasmin, in its active and inactive proenzyme, namely plasminogen, forms. Ostrich alpha(2)AP was purified using L-lysine-Sepharose chromatography, ammonium sulfate fractionation, and Super Q-650S and ostrich LBSI-Sepharose chromatographies. It revealed a M(r) of 84 K (thousand) and had one and two N-terminal amino acids in common with 11 of those of human and bovine alpha(2)AP, respectively. It showed the largest inhibitory effect on ostrich plasmin, followed by bovine trypsin and plasmin, respectively, and much less plasmin inhibition than bovine aprotinin, but much more so than human alpha(2)AP, DFP and EACA. Ostrich plasminogen was highly purified after L-lysine-Sepharose chromatography and showed a M(r) of 92 K, a total of 775 amino acids and its N-terminal sequence showed approximately 53% identity with those of human, rabbit, cat, and ox plasminogens. Ostrich plasmin, obtained by the urokinase-activation of ostrich plasminogen, revealed a M(r) of 78 K, a total of 638 amino acids, an N-terminal sequence showing two to four residues identical to five of those of human, cat, dog, rabbit, and ox plasmins, and pH and temperature optima of 8.0 and 40 degrees C, respectively. PMID- 11435136 TI - Mitogen-activated protein kinases are in vivo transducers of osmosensory signals in fish gill cells. AB - The abundance and activity of three subgroups of mitogen-activated protein (MAP) kinases, the extracellular signal regulated kinase 1 (ERK1), stress-activated protein kinase 1/ Jun N-terminal kinase (SAPK1), and stress-activated protein kinase 2/ p38 (SAPK2), were measured in gill epithelium of the euryhaline teleost Fundulus heteroclitus exposed for 1 h to 4 weeks to hyper- and hyposmotic stress. The abundance of ERK1, SAPK1 and SAPK2 was analyzed by standard Western immunodetection. MAP kinase activity is a function of phosphorylation and was measured using phospho-specific and MAP kinase subgroup-specific antibodies. The abundance of the 63 kDa fish isoform of SAPK2 increases significantly during hyper- but not hyposmotic stress while ERK1 and SAPK1 protein levels remain unchanged during both types of osmotic stress. In contrast to this small effect of osmotic stress on MAP kinase abundance, the activity of all MAP kinases decreases significantly in response to hyperosmotic stress and increases significantly during hyposmotic stress. These results demonstrate for the first time that the activity of all major MAP kinases is osmoregulated in gill epithelium of euryhaline fish. Based on these results we conclude that MAP kinases are important components of salinity adaptation and participate in osmosensory signaling pathways in gill epithelium of euryhaline fishes. PMID- 11435137 TI - Probing the mechanism of a cyanobacterial Delta9 fatty acid desaturase from Spirulina platensis C1 (Arthrospira sp. PCC 9438). AB - The initial and rate determining step in the mechanism of fatty acid desaturases has been proposed to be breakage of one of the C&z.sbnd;H bonds at the site of the incipient double bond. This has been investigated and supported for a number of eukaryotic fatty acid desaturases through the use of kinetic isotope effect experiments with deuterated substrates. In order to probe the reaction catalyzed by the cyanobacterial Delta9 desaturase and compare it to the eukaryotic desaturases, the desC gene of Spirulina platensis, strain C1 (Arthrospira sp. PCC 9438) was expressed in a desaturase mutant of baker's yeast. Kinetic isotope effects were performed by culturing yeast transformants with deuterated thia substituted stearic acids. A large kinetic isotope effect was found for the 9 position, in qualitative agreement with results from eukaryotic desaturases. PMID- 11435138 TI - The sibling species Drosophila melanogaster and Drosophila simulans differ in the expression profile of glutathione S-transferases. AB - Two major forms of glutathione S-transferase are known in Drosophila melanogaster: GST D and GST 2. In the present paper we report the existence of a third major form of glutathione S-transferase in Drosophila simulans. Induction with phenobarbital revealed a different regulation of GST between these species. Despite the fact that these two species are closely related, there was a difference in the expression profile of the enzyme implicated in the detoxification system, suggesting variations in capacity to suit their environment. PMID- 11435139 TI - High density lipoprotein binding protein of eel (Anguilla japonica) liver with specificity of binding to apoAI as a ligand. AB - High-density lipoprotein (HDL) binding protein (HBP) was isolated from the microsomal fraction of eel liver homogenate by affinity chromatography with a HDL column. After SDS-PAGE and blotting, HBP on the PVDF membrane was detected by FITC-labeled HDL and apolipoprotein AI (apoAI) as a ligand. HBP in the microsomal fraction was most abundant among microsomal, mitochondrial and cytosolic fractions. The HBP isolated by a HDL-column consisted of at least three proteins with low molecular weights of 18.5, 14.5 and 13.5 kDa; the main component was 14.5 kDa. These proteins are not products of protease digestion, as the procedure was carried out in the presence of protease inhibitors including (p-aminophenyl) methansulfonyl fluoride, 4-(2-aminoethyl)-benzenesulfonyl fluoride, pepstatin A, E-64, bestatin, leupeptin, aprotinin and EDTA. The HBP specifically bound to FITC apoAI and faintly bound or did not bind to FITC-apoAII. Furthermore, binding of HDL labeled with lipophilic fluorescence to isolated eel hepatocytes was inhibited by the antibody to apoAI, but not inhibited by the antibody to apolipoprotein AII (apoAII). These results strongly suggest that the HBP isolated from the microsomal fraction is present on the plasma membrane of eel liver and plays important roles for the lipid transport through the interaction with HDL. PMID- 11435140 TI - Thermolabile alkaline phosphatase from Northern shrimp (Pandalus borealis): protein and cDNA sequence analyses. AB - Sequence analysis of short fragments resulting from trypsin digestion of the thermolabile shrimp alkaline phosphatase (SAP) from Northern shrimp Pandalus borealis formed the basis for amplification of its encoding cDNA. The predicted protein sequence was recognized as containing the consensus alkaline phosphatase motif comprising the active site of this protein family. Protein sequence homology searches identified several eukaryote alkaline phosphatases with which the 475-amino acid SAP polypeptide revealed shares 45% amino acid sequence identity. Residues for potential metal binding seem to be conserved in these proteins. The predicted 54-kDa molecular mass of SAP is smaller than previously reported, but is consistent with our recent SDS-PAGE analysis of the native protein. Compared to its homologs, the shrimp enzyme has a surplus of negatively charged amino acids, while the relative number of prolines is lower and the frequency of aromatic residues is higher than in mesophilic counterparts. PMID- 11435141 TI - Neutral endopeptidase is expressed on the follicular granulosa cells of rabbit ovaries. AB - Neutral endopeptidase (NEP) is a zinc metallopeptidase ubiquitously distributed in various tissues in mammals. This peptidase is involved in the post-secretory metabolism of various neuropeptides and peptide hormones in vivo, such as enkephalins, bradykinin, atrial natriuretic peptide, substance P and endothelins. In this paper we show that NEP is expressed in ovaries as a 110-kDa glycosylated integral membrane protein with enzymatic properties similar to those of the kidney protein. Using immunohistochemistry, we localize the peptidase in the granulosa cells of follicles at all stages of maturation, with the exception of atretic follicles. We also observe immunoreactive staining in the epithelia that lines the blood vessels in the medulla and the surface of the ovary. The co localization of NEP and bioactive peptides known to be physiological substrates of NEP in other tissues suggests an important role for this protein in processes such as follicle maturation, ovulation, and/or regulation of ovarian blood flow, by modulating the physiological function of these peptides. PMID- 11435142 TI - Hydrocarbon-released nestmate aggression in the Argentine ant, Linepithema humile, following encounters with insect prey. AB - Argentine ants, Linepithema humile, were attacked by their nestmates following contact with a particular prey item, the brown-banded cockroach, Supella longipalpa. Contact with prey, as brief as 2 min, provoked nestmate aggression. Argentine ants contaminated with hydrocarbons extracted from S. longipalpa also released nestmate aggression behavior similar to that released by the whole prey item, confirming the involvement of hydrocarbons. In contrast to S. longipalpa, little or no nestmate aggression was induced by other ant prey from diverse taxa. A comparison of prey hydrocarbon profiles revealed that all hydrocarbons of S. longipalpa were very long chain components with 33 or more carbons, while other prey had either less, or none, of the very long chain hydrocarbons of 33 carbons or greater. We identified the hydrocarbons of S. longipalpa and some new groups of long chain hydrocarbons of L. humile. The majority of S. longipalpa hydrocarbons were 35 and 37 carbons in length with one to three methyl branches, and closely resembled two previously unidentified groups of compounds from L. humile of similar chain length. The hydrocarbons of S. longipalpa and L. humile were compared and their role in the Argentine ant nestmate recognition is discussed. PMID- 11435143 TI - Effects of pulmonary afterload on the hemodynamics after the hemi-Fontan procedure. AB - A computational fluid dynamics study based on the application of the finite volume method has been performed to investigate the effects of the pulmonary afterload on the hemodynamics after the hemi-Fontan procedure. This operation is generally used as part of a series of staged procedures to treat complex congenital malformations of the heart. It consists of re-directing the superior vena caval flow from the right atrium into the pulmonary arteries, by-passing the right ventricle while excluding the inferior caval flow from the lungs. To reproduce correctly the pulmonary afterload conditions, a simplified lumped parameter mechanical model of the pulmonary circulation has been developed and linked to the finite volume solver. In addition, the effect of a stenosis in the left pulmonary artery was also examined. In this paper the adopted methodology is presented, together with some of the preliminary results. The model has been used to simulate the local fluid dynamics for different values of the pulmonary arteriolar resistance and lung resistances, allowing a quantitative evaluation of the dissipated energy and the flow distribution into the lungs. The results show that both flow distribution into the lungs and energy dissipation after the hemi Fontan procedure are only minimally affected by the pulmonary arteriolar resistance. PMID- 11435144 TI - Platelet deposition in stagnation point flow: an analytical and computational simulation. AB - A mathematical and numerical model is developed for the adhesion of platelets in stagnation point flow. The model provides for a correct representation of the axi symmetric flow and explicitly uses shear rate to characterise not only the convective transport but also the simple surface reaction mechanism used to model platelet adhesion at the wall surface. Excellent agreement exists between the analytical solution and that obtained by the numerical integration of the full Navier--Stokes equations and decoupled conservation of species equations. It has been shown that for a constant wall reaction rate modelling platelet adhesion the maximum platelet flux occurs at the stagnation point streamline. This is in direct contrast to that found in experiment where the maximum platelet deposition occurs at some distance downstream of the stagnation point. However, if the wall reaction rate is chosen to be dependent on the wall shear stress then the analysis shows that the maximum platelet flux occurs downstream of the stagnation point, providing a more realistic model of experimental evidence. The analytical formulation is applicable to a large number of two-dimensional and axi symmetrical surface reaction flows where the wall shear stress is known a priori. PMID- 11435145 TI - A comparison of the texture of computed tomography and projection radiography images of vertebral trabecular bone using fractal signature and lacunarity. AB - The structural integrity of trabecular bone is an important factor characterizing the biomechanical strength of the vertebra, and is determined by the connectivity of the bone network and the trabeculation pattern. These can be assessed using texture measures such as the fractal signature and lacunarity from a high resolution projection radiograph. Using central sections of lumbar vertebrae we compared the results obtained from high-resolution transverse projection images with those obtained from spatially registered low-resolution images from a conventional clinical CT scanner to determine whether clinical CT data can provide useful structural information. Provided the power spectra of the CT images are corrected for image system blurring, the resulting fractal signature is similar for both modalities. Although the CT images are blurred relative to the projection images, with a consequent reduction in lacunarity, the estimated trabecular separation obtained from the lacunarity plots is similar for both modalities. This suggests that these texture measures contain essential information on trabecular microarchitecture, which is present even in low resolution CT images. Such quantitative texture measurements from CT or MRI images are potentially useful in monitoring bone strength and predicting future fracture risk. PMID- 11435146 TI - Recognition of imagined hand movements with low resolution surface Laplacian and linear classifiers. AB - EEG-based Brain Computer Interfaces (BCIs) require on-line detection of mental states from spontaneous EEG signals. In this framework, it was suggested that EEG patterns can be better detected with EEG data transformed with Surface Laplacian computation (SL) than with the unprocessed raw potentials. However, accurate SL estimates require the use of many EEG electrodes, when local estimation methods are used. Since BCI devices have to use a limited number of electrodes for practical reasons, we investigated the performances of spline methods for SL estimates using a limited number of electrodes (low resolution SL). Recognition of mental activity was attempted on both raw and SL-transformed EEG data from five healthy people performing two mental tasks, namely imagined right and left hand movements. Linear classifiers were used including Signal Space Projection (SSP) and Fisher's linear discriminant. Results showed an acceptable average correlation between the waveforms obtained with the low resolution SL and these obtained with the SL computed from 26 electrodes (full resolution SL). More importantly, satisfactorily recognition scores for mental EEG-patterns were obtained with the low-resolution surface Laplacian transformation of the recorded potentials when compared with those obtained by using full resolution SL (82%). These results demonstrated also the utility of linear classifiers for the detection of mental patterns in the BCI field. PMID- 11435147 TI - The electrochemical oxide growth behaviour on titanium in acid and alkaline electrolytes. AB - Titanium implants have a thin oxide surface layer. The properties of this oxide layer may explain the good biocompatibility of titanium implants. Anodic oxidation results in a thickening of the oxide film, with possible improved biocompatability of anodized implants. The aim of the present study was twofold: (1) firstly, to characterize the growth behaviour of galvanostatically prepared anodic oxide films on commercially pure (c.p.) titanium and (2) secondly, to establish a better understanding of the electroche0mical growth behaviour of anodic oxide on commercially pure titanium (ASTM grade 1) after changes of the electrochemical parameters in acetic acid, phosphoric acid, calcium hydroxide, and sodium hydroxide under galvanostatic anodizing mode. The oxide thickness was measured by Ar sputter etching in Auger Electron spectroscopy (AES) and the colours were estimated by an L*a*b* system (lightness, hue and saturation) using a spectrophotometer. In the first part of our study, it was demonstrated that the interference colours were useful to identify the thickness of titanium oxide. It was also found that the anodic forming voltages with slope (dV/dt) in acid electrolytes were higher than in alkaline electrolytes. Each of the used electrolytes demonstrates an intrinsically specific growth constant (nm/V) in the range of 1.4--2.78 nm/V. In the second part of our study we found, as a general trend, that an increase of electrolyte concentration and electrolyte temperature respectively decreases the anodic forming voltage, the anodic forming rate (nm/s) and the current efficiency (nm.cm(2)/C), while an increase of the current density and the surface area ratio of the anode to cathode increase the anodic forming voltage, the anodic forming rate and the current efficiency. The effects of electrolyte concentration, electrolyte temperature, and agitation speed were explained on the basis of the model of the electrical double layer. PMID- 11435148 TI - Determination of contact hip stress from nomograms based on mathematical model. AB - Nomograms are presented that enable determination of maximal stress on the hip joint weight bearing area if certain geometrical parameters of the hip and pelvis and the body weight are known. The nomograms are calculated by using previously developed mathematical models. It is demonstrated how the maximal stress on the hip joint weight bearing area is determined from the presented nomograms for a hip for which the geometrical parameters were obtained from a standard anteroposterior rentgenograph. This simple and noninvasive method may give insight into the biomechanical status of the hip which should be considered in routine surgical planning and as a part of the routine examination of the patient without the use of any additional tools. PMID- 11435149 TI - CREam of cytokinin signalling: receptor identified. AB - Cytokinins play a central role in the regulation of plant cell division and numerous developmental processes. Pleiotropic effects have made studies of this hormone difficult, and cytokinin signalling pathways have long remained elusive. The recent identification of CRE1 (a histidine kinase identical to AHK4 and WOL) as the cytokinin receptor of Arabidopsis thaliana is a landmark in cytokinin research. Mutations have been identified in CRE1, and the phenotype of loss-of function mutations sheds new light on the role of cytokinins in plant development. This article describes the experimental paths leading to receptor identification and the current interpretation of its function. PMID- 11435150 TI - Targeting detoxification pathways: an efficient approach to obtain plants with multiple stress tolerance? AB - A serious factor limiting the engineering of stress tolerance has been our ignorance about the function of stress-induced genes. A stress-activated novel aldose-aldehyde reductase was cloned from alfalfa. The ectopic expression of this gene in tobacco resulted in tolerance to oxidative stress and dehydration. Physiological analysis suggested that aldose reductase probably functions by reducing the level of reactive aldehydes. This provides a promising perspective for the development of crop plants with improved stress tolerance. PMID- 11435151 TI - Functional imaging of plants by magnetic resonance experiments. AB - Microimaging based on magnetic resonance is an experimental technique that can provide a unique view of a variety of plant physiological processes. Particularly interesting applications include investigations of water movement and spatially resolved studies of the transport and accumulation of labelled molecules in intact plant tissue. Some of the fundamental principles of nuclear and electron magnetic resonance microimaging are explained here and the potential of these techniques is shown using several representative examples. PMID- 11435167 TI - Role of short RNAs in gene silencing. AB - Recent research has revealed the existence of an elegant defence mechanism in plants and lower eukaryotes. The mechanism, known in plants as post transcriptional gene silencing, works through sequence-specific degradation of RNA. It appears to be directed by double-stranded RNA, associated with the production of short 21-25 nt RNAs, and spread through the plant by a diffusible signal. The short RNAs are implicated as the guides for both a nuclease complex that degrades the mRNA and a methyltransferase complex that methylates the DNA of silenced genes. It has also been suggested that these short RNAs might be the mobile silencing signal, a suggestion that has been challenged recently. PMID- 11435168 TI - Balance of power: a view of the mechanism of photosynthetic state transitions. AB - Photosynthesis in plants involves photosystem I and photosystem II, both of which use light energy to drive redox processes. Plants can balance the distribution of absorbed light energy between the two photosystems. When photosystem II is favoured, a mobile pool of light harvesting complex II moves from photosystem II to photosystem I. This short-term and reversible redistribution is known as a state transition. It is associated with changes in the phosphorylation of light harvesting complex II but the regulation is complex. Redistribution of energy during state transitions depends on an altered binding equilibrium between the light harvesting complex II-photosystem II and light harvesting complex II photosystem I complexes. PMID- 11435169 TI - Biomonitoring the genotoxicity of environmental factors with transgenic plants. AB - All organisms must react to constantly changing surroundings. Environmental factors are thus powerful forces continuously shaping the genomes of all species. Induced genetic changes can be followed using a biomonitor - a living organism that reacts to a given compound in the environment. A vital but challenging task is identifying organisms with which to study the influence of changing environmental conditions. Plants are especially valuable biomonitors. Here, we describe the use of transgenic plant systems to evaluate the genotoxicity of chemical and radiological compounds. We evaluate the potential of further transgene-based systems for studying somatic and germ-line mutations. PMID- 11435170 TI - Relearning our ABCs: new twists on an old model. AB - Over the past decade, the ABC model of flower development has been widely promulgated. However, correct flower-organ development requires not only the ABC genes but also the SEPALLATA genes. When the SEPALLATA genes are expressed together with the ABC genes, both vegetative and cauline leaves are converted to floral organs. Most of the ABC genes and all three SEPALLATA genes encode MADS transcription factors, which bind to DNA as dimers. Here, amendments to the ABC model are considered that incorporate both the SEPALLATA genes and the ability of MADS proteins to form higher-order complexes. PMID- 11435171 TI - Molecular recognition in thylakoid structure and function. AB - In photosynthesis, light-harvesting chlorophyll molecules are shunted between photosystems by phosphorylation of the protein to which they are bound. An anchor for the phosphorylated chlorophyll-protein complex has now been identified in the reaction centre of chloroplast photosystem I. This finding supports the idea that molecular recognition, not membrane surface charge, governs the architecture of the chloroplast thylakoid membrane. We describe a model for the chloroplast thylakoid membrane that is consistent with recent structural data that specify the relative dimensions of intrinsic protein complexes and their dispositions within the membrane. Control of molecular recognition accommodates membrane stacking, lateral heterogeneity and regulation of light-harvesting function by means of protein phosphorylation during state transitions--adaptations that compensate for selective excitation of photosystem I or photosystem II. High resolution structural description of membrane protein-protein interactions is now required to understand thylakoid structure and regulation of photosynthesis. PMID- 11435172 TI - Plasmodesmata and plant cytoskeleton. AB - Plant cell-to-cell communication is achieved by membranous conduits called plasmodesmata, which bridge the cytoplasm of neighboring cells. A growing body of immunolocalization data shows an association of the cytoskeleton machinery with plasmodesmata. The role of the cytoskeleton in the plasmodesmata-mediated transport has been well documented for virus movement. Because viruses are known to exploit existing host pathways and because the cytoskeleton is involved in intracellular trafficking, the cytoskeleton is thought to drive and target macromolecules to plasmodesmata. It is this link between plasmodesmata and the cytoskeleton that will be described here. PMID- 11435173 TI - Calcium oxalate in plants. PMID- 11435174 TI - Role of areca nut in betel quid-associated chemical carcinogenesis: current awareness and future perspectives. AB - Betel quid (BQ)-chewing is a popular oral habit with potential links to the occurrence of oral cancer. Many of the literature-based studies reveal that areca nut (AN) extract may demonstrate mutagenic and genotoxic effects, in addition to inducing preneoplastic as well as neoplastic lesions in experimental animals. Areca nut should, thus, be highly suspected as a human carcinogen. Toxicity studies relating to AN-contained polyphenols and tannins are not conclusive, with both carcinogenic and anti-carcinogenic effects being reported. The mutagenicity and genotoxicity of areca alkaloids has been detected by many short-term assays. However, their genotoxicity to oral fibroblasts and keratinocytes, the target cells of BQ, has not been identified. It would thus appear that AN toxicity is not completely due to its polyphenol, tannin and alkaloid content. The single agent which is responsible for AN carcinogenicity awaits further clarification. Reactive oxygen species produced during auto-oxidation of AN polyphenols in the BQ-chewer's saliva, are crucial in the initiation and promotion of oral cancer. Nitrosation of areca alkaloids also produces AN-specific nitrosamines, that have been demonstrated to be mutagenic, genotoxic and are capable of inducing tumors in experimental animals. Arecaidine and AN extract are further suggested to be tumor promoters. Antioxidants such as glutathione and N-acetyl-L-cysteine can potentially prevent such AN-elicited cytotoxicity. Further studies are needed to delineate the metabolism of AN ingredient and their roles in the multi-step chemical carcinogenesis, in order to enhance the success of the future chemoprevention of oral cancer and oral submucous fibrosis. PMID- 11435175 TI - Carcinoma of the hard palate treated with radiotherapy: a retrospective review of 31 cases. AB - There are few studies reporting the results of radical radiotherapy for carcinoma of the hard palate. We have examined our results of patients treated within a single institution, and assessed survival, local control and morbidity. A retrospective analysis was made on 31 patients with hard palate carcinoma treated with external beam radiotherapy at the Christie Hospital between 1990 and 1997. Twenty-six patients received radiotherapy alone and five were treated for post operative positive surgical margins. The 5-year actuarial survival rate was 55%. The actuarial 5-year local control rate was 53%, rising up to 69% after salvage surgery. Survival was 48% for squamous cell carcinomas and 63% for salivary gland carcinomas, the difference was not significant. The only significant predictor of local control was T-stage, with 80% 5-year local control of T1-2 lesions and 24% control of T3-4 lesions. N-stage was the only significant factor predicting for survival. Radiation necrosis occurred in one patient. Radical radiotherapy for carcinoma of the hard palate is safe and well tolerated. It is an effective treatment for both squamous cell carcinoma and salivary gland carcinoma. PMID- 11435176 TI - Inactivation of the p14(ARF), p15(INK4B) and p16(INK4A) genes is a frequent event in human oral squamous cell carcinomas. AB - The p14(ARF), p15(INK4B) and p16(INK4A) genes were localized to 9p21, where genetic alterations have been reported frequently in various human tumors. We performed a molecular analysis of the mechanism of inactivation in cell lines and 32 oral squamous cell carcinoma (OSCC), using deletion screening, PCR-SSCP, methylation-specific-PCR and cycle sequencing. We detected homozygous deletion of p14(ARF)-1Ebeta in 9 (26.5%), of p15(INK4B) in one (3.1%), and of p16(INK4A) in 22 (56.3%) tumor samples. Three mutations were detected in the p16(INK4A) genes. We detected aberrant methylation of the p14(ARF) genes in 14 (43.8%), of the p15(INK4B) gene in 9 (28.1%), and of the p16(INK4A) gene in 16 (50.0%) tumor samples. Altogether, 87.5% of the samples harbored at least one of the alterations in the p14(ARF), p15(INK4B), and p16(INK4A) genes, indicating that the frequent inactivation of these genes may be an important mechanism during OSCC development. PMID- 11435177 TI - Impact of localized treatment in reducing risk of progression of low-grade oral dysplasia: molecular evidence of incomplete resection. AB - Currently, there is no consensus on the appropriate treatment for low-grade oral dysplasia. This is mainly due to the difficulty in predicting outcome for this heterogeneous group of lesions. In this study, we constructed a detailed clinical history of 66 mild and moderate dysplasias in order to determine how treatment affected outcome, and to evaluate the effect of treatment on lesions with different genetic profiles, which are defined by patterns of loss of heterozygosity (LOH) associated with low, intermediate and high risk of progression [Clin. Cancer Res., 6, 357-62, 2000]. The results showed that although treatment guided by clinical removal of leukoplakia reduced cancer progression risk in all three risk groups, the amount of reduction in our study group did not reach statistical significance. To assess whether completeness of lesion removal was a major factor in recurrence, repeat biopsies at the primary sites were analyzed for persistent LOH status on chromosomes 3p, 4q, 8p, 9p, 11q, 13q and 17p. Strikingly, eight of 17 cases judged clinically removed contained the same molecular clones in the initial and subsequent biopsies, suggesting incomplete removal. When molecular information was included in the assessment of lesion removal, treatment significantly reduced the risk of progression for cases with intermediate (P=0.043) and high risk (P=0.001) genetic profiles, but not cases with low-risk profiles. A 9.1-fold decrease in progression risk was observed for those with high-risk profile. Altogether, these data suggest the use of molecular profiles to guide the treatment of low-grade dysplasia. Our data also suggest that currently an inadequate margin may in part be responsible for the high rate of recurrence, especially in high-risk lesions. PMID- 11435178 TI - Factors associated with smoking and alcohol consumption following treatment for head and neck cancer. AB - The study aim was to investigate the correlates of smoking and alcohol drinking in post-therapeutic head and neck (H&N) cancer patients. A cross-sectional design was used with a sample of 191 patients. Data were collected by interview and chart review. Multiple logistic regression analysis was used to evaluate the correlates of dependent variables. Higher education, living with one's partner, later stage, laryngeal site and having surgery or combined therapy were associated with decreased odds for smoking. There was a significant trend for decreasing odds for smoking with increasing stage. Male gender, stage I disease and longer time since treatment were associated with increased odds for drinking alcohol. There was a significant trend for increased odds for drinking with increased time since treatment. These findings suggest that smoking and alcohol drinking have different patterns of associated variables in post-therapeutic H&N cancer patients, which has important implications for intervention design. PMID- 11435179 TI - In situ staining with DNA-binding fluorescent dye, Hoechst 33258, to detect microorganisms in the epithelial cells of oral leukoplakia. AB - This study was performed to investigate the presence of microorganisms in the epithelial cells of leukoplakia. Frozen sections of 20 specimens of leukoplakia were stained with DNA-binding bisbenzimide Hoechst 33258. As a control, 20 specimens of normal oral mucosa and five specimens of normal skin were used. In all preparations of leukoplakia, small granular fluorescing structures were observed within the cytoplasm of the epithelial cells, predominantly within the cytoplasm of prickle cells, although the amount of the granular structures varied between specimens, layers of the epithelium and even areas of the epithelium within a single section. Less granular structures were observed, or none at all, in the cytoplasm of the epithelial cells of normal mucosa. No structures were observed in the cytoplasm of the epithelium of skin. The results in this study strongly suggest that microorganisms are present in the epithelial cells of oral mucosa, and that they are closely associated with the development of oral leukoplakia. It is postulated that the microorganisms in the epithelial cells could be bacteria, particularly mycoplasmas. PMID- 11435180 TI - Characterization of four cell lines derived from a human malignant fibrous histiocytoma of the maxillary sinus. AB - We have established four cell lines from a human malignant fibrous histiocytoma. Each cell line had human aneuploid karyotype and DNA aneuploidy. Cells in all lines expressed CD13, CD68 and vimentin but lacked CD11, CD14, CD15, CD16, CD45, HLA class II and other mesenchymal and epithelial markers such as desmin, alpha smooth muscle, myoglobin, S-100 protein, and cytokeratin. None of the cells expressed surface IgG or C3 receptor, nor did any of them phagocytose latex particles. The cells reacted with an antibody for prolyl-4-hydroxylase, but no collagen (types I, II, III, or IV) was detected in any of the cell lines. The homogenates of all cell lines had cyclic nucleotide phosphodiesterase 3 activity. Two cell lines produced granulocyte colony-stimulating factor (G-CSF), interleukin (IL)-1alpha, IL-6 and tumor necrosis factor alpha, another line produced G-CSF, IL-1alpha, IL-6 and platelet-derived growth factor (PDGF)-AB, and the remaining cell line produced only PDGF-AB. None of the cells produced transforming growth factor-alpha. The results indicated that the cell lines were immunophenotypically similar to primitive mesenchymal cells. PMID- 11435181 TI - 5-Fluorouracil induces autophagic degeneration in rat oral keratinocytes. AB - In this study, we investigated the effect of 5-fluorouracil (5-FU) on the keratinocytes of oral epithelium. Female Lewis rats were given 5-FU i.v. and were killed 12, 24 or 36 h after injection. The buccal mucosa was dissected. The number of nuclei with DNA strand breaks and the total number of nuclei per volume of the epithelial basal cell layer was estimated using terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP)-biotin nick end labeling. Epithelial cells were analysed by flow cytometry, transmission electron microscopy and a dye exclusion test. The number of cells with DNA strand breaks increased in 5-FU treated rats. Flow cytometry showed a decrease in cell size and an increase in granularity with increasing doses of 5-FU. Dye exclusion gave no indication of degenerate cell membranes. By transmission electron microscopy, the cells showed evidence of degeneration, shrinkage and loss of cell-to-cell contact. Vacuolation was extensive and, in contrast to apoptotic cells, nuclear chromatin condensation seemed to occur centrally in the nuclei. The results show that 5-FU treatment in vivo induces alterations in rat oral keratinocytes that are consistent with autophagic degeneration. PMID- 11435182 TI - Imaging of mandibular trauma: ROC analysis. AB - OBJECTIVES: To compare the diagnostic efficacies of panoramic radiographs, mandibular trauma series, and digitized radiographs for detection of fractures of the mandible. METHODS: Fractures were induced using blunt trauma in 25 cadaver mandibles. Panoramic radiographs, digitized radiographs, and mandibular series comprising an anteroposterior view, two lateral oblique, and a reverse Towne's were used. Six observers recorded their diagnoses using a five-point confidence rating scale. The data were analyzed using receiver operating characteristic (ROC) curve analysis. RESULTS: Significant differences based on imaging modalities were found (p < 0.0015) in the area under the curves (A(z)): panoramic radiograph, 0.8762; mandibular series, 0.7521; panoramic plus anteroposterior radiographs combination, 0.8886; and digitized mandibular series, 0.7723. No observer-based differences were noted. Condylar and coronoid fractures were difficult to detect than those in other areas of the mandible (p < 0.033). Intra- and inter-observer agreements were high (kappa(w) = 0.81 and 0.76, respectively). CONCLUSIONS: Panoramic radiographs are adequate for detection of mandibular fractures. Addition of an anteroposterior view augments diagnostic accuracy. PMID- 11435183 TI - Combination of Goldman risk and initial cardiac troponin I for emergency department chest pain patient risk stratification. AB - BACKGROUND: Accurate identification of low-risk emergency department (ED) chest pain patients who may be safe for discharge has not been well defined. Goldman criteria have reliably risk-stratified patients but have not identified any subset safe for ED release. Cardiac troponin I (cTnI) values have also been shown to risk-stratify patients but have not identified a subset safe for ED release. OBJECTIVE: To test the hypothesis that ED chest pain patients with a Goldman risk of < or =4% and a single negative cTnI (< or =0.3 ng/mL) at the time of ED presentation would be safe for discharge [<1% risk for death, acute myocardial infarction (AMI), revascularization]. METHODS: A prospective cohort study was performed in which consecutive ED chest pain patients were enrolled from July 1999 to November 2000. Data collected included patient demographics, medical and cardiac history, electrocardiogram, and creatine kinase-MB and cTnI. Goldman risk stratification score was calculated while patients were still in the ED. Hospital course was followed daily. Telephone follow-up occurred at 30 days. The main outcome was death, AMI, or revascularization (percutaneous transluminal coronary angioplasty/stents/coronary artery bypass grafting) within 30 days. RESULTS: Of 2,322 patients evaluated, 998 had both a Goldman risk < or =4% and a cTnI < or =0.3 ng/mL. During the initial hospitalization, 37 patients met the composite endpoint (3.7%): 6 deaths (0.7%), 17 AMIs (1.7%), 18 revascularizations (1.8%). Between the time of hospital discharge and 30-day follow-up, 15 patients met the composite endpoint: 4 deaths (0.4%), 6 AMIs (0.6%), and 5 revascularizations (0.5%). Overall, 49 patients met the composite endpoint (4.9%; 95% CI = 3.6% to 6.2%): 10 deaths (1.0%; 95% CI = 0.4% to 1.6%); 23 AMIs (2.3%; 95% CI = 1.4% to 3.2%), and 23 revascularizations (2.3%; 95% CI = 1.4% to 3.2%) within 30 days of presentation. CONCLUSIONS: The combination of two risk stratification modalities for ED chest pain patients (Goldman risk < or =4% and cTnI < or =0.3 ng/mL) did not identify a subgroup of chest pain patients at <1% risk for death, AMI, or revascularization within 30 days. PMID- 11435184 TI - Characteristics and outcomes of young adults who present to the emergency department with chest pain. AB - BACKGROUND: Most studies of emergency department (ED) chest pain patients exclude patients <30-40 years old. As a result, the clinical course of these patients is poorly described. OBJECTIVE: To study the clinical characteristics, hospital course, and 30-day outcomes of ED chest pain patients <40 years old. The hypothesis was that patients <40 years old without a cardiac history and with normal electro-cardiograms (ECGs) or no cardiac risk factors would be at a <1% risk for acute coronary syndromes (ACSs) and 30-day adverse cardiovascular (CV) events. METHODS: This was a prospective cohort study of non-cocaine-using ED patients, 24-39 years old, who received an ECG for chest pain between July 9, 1999, and October 23, 2000. Structured data collection at presentation included demographics, chest pain description, history, laboratory, and ECG data. Hospital course was followed daily. Thirty-day follow-up was performed by telephone. The main outcomes were discharge diagnosis and 30-day adverse CV events [acute myocardial infarction (AMI), death, percutaneous intervention (PCI), or coronary artery bypass grafting (CABG)]. RESULTS: A total of 487 patients presented 527 times and comprised the study group. Patients were most often 30-39 years old (71%), female (60%), and African-American (73%). Thirty-two percent were admitted. Five hundred seven of 527 patient visits (96%) had 30-day follow-up. Patients had the following cardiac risk factors: tobacco, 37%; hypertension, 22%; family history, 19%; diabetes mellitus, 6%; cholesterol, 6%; prior angina, 3%; known coronary artery disease, 3%; and prior AMI, 2%. Patients usually had unremarkable ECGs (61% normal, 98% nonischemic). Overall, 11 of 527 patients had adverse CV events (2.1%; 95% CI = 0.9% to 3.3%): 8 AMIs (1.5%), 4 deaths (0.8%), 5 PCIs (0.9%), and no CABG. Twenty-five patients had a final diagnosis of ACS (4.7%; 95% CI = 2.9% to 6.5%). The incidence of ACS in the 210 patients without a cardiac history and without cardiac risk factors was 0.5% (95% CI = 0% to 1.4%). At 30 days, none of these 210 patients had AMI, PCI, CABG, or death (0%, 95% CI = 0% to 1.4%). The incidence of ACS in the 312 patients with normal ECGs and a negative cardiac history was 0.3% (95% CI = 0% to 0.9%). At 30 days, there was no AMI, PCI, or CABG in these 312 patients, and one patient with metastatic cancer died (adverse CV event 0.3%, 95% CI = 0% to 0.9%). CONCLUSIONS: Although young patients, as a whole, have a 4.7% risk of ACSs and a 2.1% risk of adverse CV events at 30 days, those without known cardiac disease or any cardiac risk factors had a <1% risk of ACSs and were free from adverse CV events over 30 days. Likewise, young patients without a cardiac history and with a normal ECG had a <1% risk of ACSs and adverse CV events at 30 days. It may be reasonable to expedite outpatient management and limit unnecessary admissions in these cohorts. PMID- 11435185 TI - Dedicated asthma center improves the quality of care and resource utilization for pediatric asthma: a multicenter study. AB - OBJECTIVES: To determine the relative effectiveness of pediatric asthma care among patients treated by a dedicated asthma center (AC) vs children who use the emergency department (ED) as a site of primary asthma care. METHODS: A retrospective case-control design was used. A random sample of AC cases was selected from a designated comprehensive AC over a 12-month period. Concurrent ED control patients were identified from all cases of pediatric asthma from five urban hospitals based on two or more ED visits. Cases and controls were matched (1:2) based on age and National Heart, Lung, and Blood Institute (NHLBI) asthma severity of illness classification. A telephone survey was administered to the caregivers of all enrolled patients in the study sample. RESULTS: Four elements of pediatric asthma care were examined: quality, access, hospital utilization, and functional impact of disease. Demographic data were similar between the ED cases and the AC controls. In terms of quality of care, the AC patients were more likely to use maintenance antiinflammatory medications, 60.2% vs 22.5% (OR = 5.3; 95% CI = 2.9 to 9.7) and more likely to be taking medications at school, 71.4% vs 48.1% (OR = 2.7; 95% CI = 1.5 to 4.7). In terms of access to care, the AC families were more likely to have a physician to call to assist with outpatient management, 98.2% vs 65.0% (OR = 25.3; 95% CI = 9.0 to 76.9). Frequent ED utilization (> or = 1 visit/month) was less likely in the AC patients, 9.2% vs 22.0% (OR = 0.35; 95% CI = 0.16 to 0.79) and school absenteeism was lower as well (9.5 +/- 6.7 days vs 16.6 +/- 10.3, p < 0.001). Additionally, the caregivers of the AC patients missed fewer workdays (4.7 +/- 2.8 vs 7.4 +/- 4.1; p = 0.03). CONCLUSIONS: Significant disparities in quality, access, resource utilization, and functional impact exist between AC and ED patients. Emergency physicians have a unique opportunity to improve the public health by directing ED patients toward pediatric AC treatment. PMID- 11435186 TI - Risk factors for infection in patients with traumatic lacerations. AB - BACKGROUND: Most of our knowledge of laceration management comes from studies in animal models or patients with closure of sterile postoperative surgical incisions. Traumatic laceration management has not been well studied. OBJECTIVE: To determine which characteristics of traumatic lacerations were associated with the development of wound infection. METHODS: A cross-sectional study of consecutive patients with traumatic lacerations repaired over a four-year period was conducted. Structured closed-question data sheets were prospectively completed at the time of laceration repair and suture removal. Infection was determined at the time of suture removal. Multivariate modeling was used to determine the adjusted odds ratio (OR) of infection. RESULTS: Five thousand five hundred twenty-one patients were enrolled; 195 patients developed an infection (3.5%). An increased likelihood of wound infection was associated with age (adjusted OR per year, 1.01; 95% CI = 1.0 to 1.02); history of diabetes mellitus (adjusted OR 6.7; 95% CI = 1.7 to 26.4); laceration width (adjusted OR 1.05 per mm; 95% CI = 1.02 to 1.08); and presence of foreign body (adjusted OR 2.6; 95% CI = 1.3 to 5.2). Laceration location on the head/neck was associated with a decreased risk of infection (adjusted OR 0.28; 95% CI = 0.18 to 0.45). CONCLUSIONS: Both patient and wound characteristics of traumatic lacerations have an influence on the likelihood of infection. This knowledge may be valuable for determining whether various methods of wound cleansing, debridement, and repair can improve the outcome of patients with traumatic lacerations. PMID- 11435187 TI - Do elder emergency department patients and their informants agree about the elder's functioning? AB - OBJECTIVE: To compare elder patients' and their informants' ratings of the elder's physical and mental function measured by a standard instrument, the Medical Outcomes Study Short Form 12 (SF-12). METHODS: This was a randomized, cross-sectional study conducted at a university-affiliated community teaching hospital emergency department (census 65,000/year). Patients >69 years old, arriving on weekdays between 10 AM and 7 PM, able to engage in English conversation, and consenting to participate were eligible. Patients too ill to participate were excluded. Informants were people who accompanied and knew the patient. Elder patients were randomized 1:1 to receive an interview or questionnaire version of the SF-12. The questionnaire was read to people unable to read. Two trained medical students administered the instrument. The SF-12 algorithm was used to calculate physical (PCS) and mental (MCS) component scores. Oral and written versions were compared using analysis of variance. The PCS and MCS scores between patient-informant pairs were compared with a matched t-test. Alpha was 0.05. RESULTS: One hundred six patients and 55 informants were enrolled. The patients' average (+/-SD) age was 77 +/- 5 years; 59 (56%; 95% CI = 46% to 65%) were women. There was no significant difference for mode of administration in PCS (p = 0.53) or MCS (p = 0.14) scores. Patients rated themselves higher on physical function than did their proxies. There was a 4.1 (95% CI = 99 to 7.2) point difference between patients' and their proxies' physical component scores (p = 0.01). Scores on the mental component were quite similar. The mean difference between patients and proxies was 0.49 (95% CI = 3.17 to 4.16). The half point higher rating by patients was not statistically significant (p = 0.79). CONCLUSIONS: Elders' self-ratings of physical function were higher than those of proxies who knew them. There was no difference in mental function ratings between patients and their proxies. Switching from informants' to patients' reports in evaluating elders' physical function in longitudinal studies may introduce error. PMID- 11435188 TI - Online medical control and initial refusal of care: does it help to talk with the patient? AB - The use of online medical control (OLMC) for initial refusal of care is time consuming and has medical-legal risks. OBJECTIVES: This study attempted to answer the following: Does physician-patient communication (PPC) increase the rate of transport and at what cost in terms of increased physician time? Do altered mental status (AMS) patients agree to transport more frequently and do they require more physician time? Can senior emergency medicine resident (RES) and emergency medicine faculty (FAC) physicians be equally efficient in handling refusal calls? METHODS: The study evaluated a retrospective cohort for six months at a single base station, university hospital. Online medical control audiotapes and written records of radio and telephone communications were reviewed. RESULTS: One hundred forty-seven refusal cases were analyzed, PPC was used 70 times, and 37 patients were transported. Twenty-four of 70 (34%) patients with PPC agreed to be transported, while ten of 77 (13%) patients without PPC agreed to be transported (p = 0.002). Sixteen of 30 (53%) patients with AMS were transported compared with 21 of 117 (18%) patients with normal mental status (p = 0.00007). Call times were longer with PPC utilization [406.3 sec PPC vs 230.1 sec no PPC (p < 0.001)] and with AMS patients [411.2 sec AMS vs 289.1 sec no AMS (p = 0.028)]. The RES and FAC physicians did not differ in transport rates [21% RES vs 26% FAC (p = 0.612)] and call times [329.4 sec RES vs 310.4 sec FAC (p = 0.659)]. CONCLUSIONS: Although time-consuming, the use of PPC is associated with more patients' agreeing to be transported. Patients with AMS are transported more frequently and they use more physician time. Emergency medicine RES and FAC physicians have equal efficiency and efficacy in handling these calls. PMID- 11435189 TI - Redefining emergency medicine procedures: Canadian competence and frequency survey. AB - OBJECTIVE: To redefine the Royal College of Physicians and Surgeons (RCPS) procedural skills list for Canadian emergency medicine (EM) residents through a national survey of EM specialists to determine procedural performance frequency and self-assessment of competence. METHODS: The survey instrument was developed in three phases: 1) an EM program directors survey identified inappropriate or dated procedures, endorsing 127 skills; 2) a search of EM literature added 98 skills; and 3) an expert panel designed the survey instrument and finalized a list of 150 skills. The survey instrument measured the frequency of procedure performance or supervision, self-reported competence (yes/no), and endorsement of one of four training levels for each skill: undergraduate (UG), postgraduate (PG), knowledge only, or unnecessary (i.e., too infrequently performed to maintain competence). RESULTS: All 289 Canadian EM specialists were surveyed by mail; 231 (80%) responded, 221 completed surveys, and 10 were inactive. More than 60% reported competence in 125 (83%) procedures, and 55 procedures were performed at least three times a year. The mean competence score was 121 (SD +/- 17.7, median = 122) procedures. Competence score correlation with patient volume was r = 0.16 (p = 0.02) and with hours worked was r = 0.19 (p = 0.01). Competence score was not associated with year or route (residency vs grandfather) of certification. Each procedure was assigned to a training level using response consensus and decision rules (UG: 1%; PG: 82%; unnecessary: 17%). CONCLUSIONS: A survey of EM clinicians reporting competence and frequency of skill performance defined 127 procedural skills appropriate for Canadian RCPS postgraduate training and EM certification. PMID- 11435190 TI - Clinical pearls: shoulder pain. PMID- 11435191 TI - Patient and provider attitudes toward commercial television film crews in the emergency department. AB - One of the most successful reality-based television shows is The Learning Channel's "Trauma-Life in the ER," in which patients are filmed prior to being consented and camera crews are intimately intertwined in resuscitations. OBJECTIVE: To assess provider and patient attitudes regarding privacy and satisfaction during filming. METHODS: This was a prospective survey of patients and providers working in an urban, county teaching emergency department (ED). Scores of visual analog scales (VASs) for satisfaction, privacy, and willingness to return to the ED were accessed. RESULTS: Eighty patients, 39 physicians and 39 nurses, were interviewed. On a 10-cm VAS where 0 = no invasion of patient privacy and 10 = extreme privacy invasion, the mean (+/-SD) rating by physicians was 5.4 (+/-2.6), by nurses was 4.9 (+/-3.1), and by patients was 2.3 (+/-3.2). There was no significant difference between physician and nurse ratings (p = 0.69), but patients rated invasion of privacy significantly lower (p < 0.0001). Filmed patients rated significantly higher invasion of privacy (3.8 +/- 4.1) than patients not filmed or those unsure whether they had been filmed (1.6 +/- 2.5); p < 0.01). On a 10-cm VAS where -5 = extreme decrease in satisfaction and 5 = extreme increase in satisfaction, the mean rating by physicians was -0.25 (+/ 1.6), by nurses was -0.32 (+/-1.3), and by patients was 0.02 (+/-1.4). There was no statistical difference between the satisfaction levels of providers and patients (p = 0.19). CONCLUSIONS: Providers rated invasion of patient privacy higher than patients rated their own invasion of privacy. Patients who were filmed rated invasion of privacy higher than patients who were not filmed. Filming had no significant effect on the satisfaction of providers or patients. PMID- 11435192 TI - Research guidelines: it's all about integrity. PMID- 11435193 TI - Pediatric emergency medicine: making SAEM a comfortable home. Society for Academic Emergency Medicine. PMID- 11435194 TI - Another attempt at easing chest pain: ours and theirs. PMID- 11435195 TI - How "low" can we go: assessing risk in chest pain patients. PMID- 11435196 TI - Guidelines for research in cooperation with biomedical industry organizations. PMID- 11435197 TI - The golden hour: scientific fact or medical "urban legend"? AB - The term "golden hour" is commonly used to characterize the urgent need for the care of trauma patients. This term implies that morbidity and mortality are affected if care is not instituted within the first hour after injury. This concept justifies much of our current trauma system. However, definitive references are generally not provided when this concept is discussed. It remains unclear whether objective data exist. This article discusses a detailed literature and historical record search for support of the "golden hour" concept. None is identified. PMID- 11435198 TI - Adrenal insufficiency in critically ill emergency department patients: a Taiwan preliminary study. AB - OBJECTIVE: Unrecognized adrenal insufficiency can have serious consequences in critically ill emergency department (ED) patients. This prospective pilot study of adrenal function in patients with severe illness was undertaken to determine the prevalence of adrenal dysfunction and any relation to prior herbal drug use. METHODS: In a high-volume urban tertiary care ED, adult patients with sepsis or acute myocardial infarction (AMI) were eligible for the study. Over a two-month period, a convenience sample was enrolled by the authors on arrival to the ED. Inclusion criteria were systemic inflammatory response syndrome (SIRS) criteria plus evidence of at least one organ dysfunction or cardiac marker plus electrocardiogram-proven AMI. Exclusion criteria included known corticosteroid use. Serum cortisol was measured on arrival and for those patients with a level of <15 microg/dL (<414 nmol/L), an adrenocorticotropic hormone (ACTH) stimulation test was performed. RESULTS: Of the 30 enrolled patients, 23 (77%) were suffering from severe sepsis and the other seven (23%) had an AMI. Thirteen of the 30 patients (43%; 95% CI = 25% to 65%) had serum cortisol levels of <15 microg/dL, consistent with adrenal insufficiency, nine with severe sepsis and four with an AMI. Eight (62%; 95% CI = 32% to 86%) of the 13 patients with low cortisol levels reported using herbal medications, while only two (12%; 95% CI = 1% to 36%) of the 17 with normal cortisol levels reported taking herb drugs (p = 0.01). Only two (15%; 95% CI = 2% to 45%) of the patients with low cortisol levels failed their corticotropin stimulation test, suggestive of true adrenocortical insufficiency. Both reported using herbal preparations. CONCLUSIONS: These results indicate that adrenal dysfunction is common among a group of critically ill patients seen in this Taiwanese ED. Moreover, the use of herbal drugs was high in the patients with low serum cortisols. Further studies are required to both confirm these findings and clarify whether a number of herbal medications contain corticosteroids. PMID- 11435199 TI - Evaluation of a residency program's experience with a one-week emergency medicine resident rotation at a medical liability insurance company. AB - INTRODUCTION: The authors' residency program implemented a one-week rotation at the office of a medical liability insurance company. Residents examined 30 closed malpractice claims cases and sat in on settlement discussions. OBJECTIVE: To review the residents' evaluations of their experiences and to determine whether this was a worthwhile addition to the emergency medicine (EM) residency curriculum. METHODS: This was a five-year retrospective study that reviewed residents' annual evaluations from 1994 to 1999 regarding the medical liability rotation. A five-point scale was used to score specific categories in the rotation and an open-ended section was used to collect general comments. RESULTS: A total of 179 resident evaluations were reviewed. The quality of teaching ranked in the 80th percentile, the clinical caseload ranked in the 85th percentile, and level of responsibility ranked in the 79th percentile for all EM rotations. Specific comments included "All MDs should do this in their training"; "Quite an eye opener"; and "Good exposure to legal aspects of EM." CONCLUSIONS: Overall, EM residents found the one-week rotation to be invaluable and a good learning experience. This rotation ranked above average when compared with all of our other EM residency rotations. PMID- 11435200 TI - Hypoglycemia in cocaine-intoxicated mice. PMID- 11435201 TI - DNA damage and cell cycle checkpoints in hyperoxic lung injury: braking to facilitate repair. AB - The beneficial use of supplemental oxygen therapies to increase arterial blood oxygen levels and reduce tissue hypoxia is offset by the knowledge that it injures and kills cells, resulting in increased morbidity and mortality. Although many studies have focused on understanding how hyperoxia kills cells, recent findings reveal that it also inhibits proliferation through activation of cell cycle checkpoints rather than through overt cytotoxicity. Cell cycle checkpoints are thought to be protective because they allow additional time for injured cells to repair damaged DNA and other essential molecules. During recovery in room air, the lung undergoes a burst of proliferation to replace injured and dead cells. Failure to terminate this proliferation has been associated with fibrosis. These observations suggest that growth-suppressive signals, which inhibit proliferation of injured cells and terminate proliferation when tissue repair has been completed, may play an important role in the pulmonary response to hyperoxia. Because DNA replication is coupled with DNA repair, activation of cell cycle checkpoints during hyperoxia may be a mechanism by which cells protect themselves from oxidant genotoxic stress. This review examines the effect of hyperoxia on DNA integrity, pulmonary cell proliferation, and cell cycle checkpoints activated by DNA damage. PMID- 11435202 TI - Lung disease in cystic fibrosis: is airway surface liquid composition abnormal? PMID- 11435203 TI - X-ray microanalysis of airway surface liquid collected in cystic fibrosis mice. AB - The airway surface liquid (ASL) that lines the airway surface epithelium plays a major role in airway antibacterial defense and mucociliary transport efficiency, two key factors in cystic fibrosis (CF) disease. A major difficulty is to collect ASL in native conditions without stimulation or alteration of the underlying airway epithelium. Using a cryoprobe specifically adapted to collect native ASL from the tracheal mouse surface, we analyzed by X-ray microanalysis the complete ASL and plasma ion content in Cftr(tm1Hgu)/Cftr(tm1Hgu) mice compared with that in control littermates. ASL ion content from eight Cftr(tm1Hgu)/Cftr(tm1Hgu) mice and eight control littermates did not appear significantly different. The mean (+/-SE) concentrations were 2,352 +/- 367 and 2,058 +/- 401 mmol/kg dry weight for Na, 1,659 +/- 272 and 1,448 +/- 281 mmol/kg dry weight for Cl, 357 +/- 57 and 337 +/- 38 mmol/kg dry weight for S, 1,066 +/- 220 and 787 +/- 182 mmol/kg dry weight for K, 400 +/- 82 and 301 +/- 58 mmol/kg dry weight for Ca, 105 +/- 31 and 105 +/- 20 mmol/kg dry weight for Mg, 33 +/- 15 and 29 +/- 9 mmol/kg dry weight for P in non-CF and CF mice, respectively. This cryotechnique appears to be a promising technique for analyzing the complete elemental composition of native ASL in CF and non-CF tissues. PMID- 11435204 TI - Hypoxic pulmonary vasoconstriction: a multifactorial response? PMID- 11435205 TI - Hypoxic release of calcium from the sarcoplasmic reticulum of pulmonary artery smooth muscle. AB - The hypoxic constriction of isolated pulmonary vessels is composed of an initial transient phase (phase 1) followed by a slowly developing increase in tone (phase 2). We investigated the roles of the endothelium and of intracellular Ca2+ stores in both preconstricted and unpreconstricted intrapulmonary rabbit arteries when challenged with hypoxia (PO2 16-21 Torr). Removing the endothelium did not affect phase 1, but phase 2 appeared as a steady plateau. Removing extracellular Ca2+ had essentially the same effect as removing the endothelium. Depletion of sarcoplasmic reticulum Ca2+ stores with caffeine and ryanodine abolished the hypoxic response. Omitting preconstriction reduced the amplitude of the hypoxic response but did not qualitatively affect any of the above responses. We conclude that hypoxia releases intracellular Ca2+ from ryanodine-sensitive stores by a mechanism intrinsic to pulmonary vascular smooth muscle without the need for Ca2+ influx across the plasmalemma or an endothelial factor. Our results also suggest that extracellular Ca2+ is required for the release of an endothelium-derived vasoconstrictor. PMID- 11435206 TI - Fas and apoptosis in the alveolar epithelium: holes in the dike? PMID- 11435207 TI - Recombinant human Fas ligand induces alveolar epithelial cell apoptosis and lung injury in rabbits. AB - This study investigated whether recombinant human soluble Fas ligand (rh-sFasL) induces apoptosis of primary type II pneumocytes in vitro and lung injury in vivo. Type II cells isolated from normal rabbit lung expressed Fas on their surface and became apoptotic after an 18-h incubation with rh-sFasL. Fas expression in normal rabbit lungs was localized by immunohistochemistry to alveolar and airway epithelia and alveolar macrophages. The administration of 10 microg of rh-sFasL into the right lungs of rabbits resulted 24 h later in both significantly more bronchoalveolar lavage fluid total protein and significantly more tissue changes compared with those in the left lungs, which received rh sFasL plus Fas:Ig (a fusion protein that binds and blocks sFasL). Tissue changes included thickening of the alveolar walls, neutrophilic infiltrates, apoptotic (terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling-positive) cells in the alveolar walls, and increased expression of interleukin-8 by alveolar macrophages (as determined by immunohistochemistry). We conclude that the alveolar epithelium of normal rabbits expresses Fas and that sFasL induces lung injury and inflammation in rabbits. PMID- 11435208 TI - Anti-neutrophil chemokine preserves alveolar development in hyperoxia-exposed newborn rats. AB - Inflammation may contribute to lung injury and impaired alveolar development in bronchopulmonary dysplasia. We treated hyperoxia-exposed newborn rats with antibodies to the neutrophil chemokine cytokine-induced neutrophil chemoattractant-1 (CINC-1) during 95% O2 exposure to reduce adverse effects of hyperoxia-induced inflammation on lung development. Rats were exposed at birth to air, 95% O2, or 95% O2 + anti-CINC-1 (injected on days 3 and 4). Bromodeoxyuridine (BrdU) was injected 6 h before death. Anti-CINC-1 treatment improved weight gain but not survival at day 8. Anti-CINC-1 reduced bronchoalveolar lavage neutrophils at day 8 to levels equal to air controls. Total detectable lung CINC-1 was reduced to air control levels. Lung compliance was improved by anti-CINC-1, achieving air control levels in the 10-microg anti CINC-1 group. Anti-CINC-1 preserved proliferating cell nuclear antigen expression in airway epithelium despite 95% O2 exposure. BrdU incorporation was depressed by hyperoxia but preserved by anti-CINC-1 to levels similar to air control. Alveolar volume and surface density were decreased by hyperoxia but preserved by anti-CINC 1 to levels equal to air control. Blockade of neutrophil influx in newborns may avert early lung injury and avoid alveolar developmental arrest that contributes to bronchopulmonary dysplasia. PMID- 11435209 TI - Endocytosed SP-A and surfactant lipids are sorted to different organelles in rat type II pneumocytes. AB - Intracellular transport of endocytosed surfactant protein A (SP-A) and lipid was investigated in isolated rat type II cells. After internalization, SP-A and lipid are taken up via the coated-pit pathway and reside in a common compartment, positive for the early endosomal marker EEA1 but negative for the lamellar body marker 3C9. SP-A then recycles rapidly to the cell surface via Rab4-associated recycling vesicles. Internalized lipid is transported toward a Rab7-, CD63-, 3C9 positive compartment, i.e., lamellar bodies. Inhibition of calmodulin led to inhibition of uptake and transport out of the EEA1-positive endosome and thus of resecretion of both components. Inhibition of intravesicular acidification (bafilomycin A1) led to decreased uptake of both surfactant components. It inhibited transport out of early endosomes for lipid only, not for SP-A. We conclude that in type II cells, endocytosed SP-A and lipid are transported toward a common early endosomal compartment. Thereafter, both components dissociate. SP A is rapidly recycled to the cell surface and does not enter classic lamellar bodies. Lipid is transported toward lamellar bodies. PMID- 11435210 TI - C-type natriuretic peptide system in fetal ovine pulmonary vasculature. AB - C-type natriuretic peptide (CNP) is a recently described endothelium-derived relaxing factor. CNP relaxes vascular smooth muscle and inhibits smooth muscle proliferation by binding to natriuretic peptide receptor (NPR) type B (NPR-B) and producing cGMP. Lung parenchyma and fifth-generation pulmonary arteries (PA) and veins (PV) were isolated from late-gestation fetal lambs. All three types of NPR mRNA were detected in PA and PV by RT-PCR. CNP and NPR-B immunostaining was positive in pulmonary vascular endothelium and medial smooth muscle. CNP concentration-response curves of PA and PV were compared with those of atrial natriuretic peptide (ANP) by use of standard tissue bath techniques. CNP relaxed PV significantly better than PA. ANP relaxed PA and PV equally, but ANP relaxed PA significantly better than CNP. Pretreating PA and PV with natriuretic peptide receptor blocker (HS-142-1) or cGMP-dependent protein kinase inhibitor Rp-beta phenyl-1- N2-etheno-8-bromoguanosine 3',5'-cyclic monophosphorothionate significantly inhibited the CNP relaxation response, indicating that the response was mediated through the NPR-cGMP pathway. We conclude that CNP is important in mediating pulmonary venous tone in the fetus. PMID- 11435211 TI - Upregulation of lung soluble guanylate cyclase during chronic hypoxia is prevented by deletion of eNOS. AB - Hypoxia upregulates endothelial (e) nitric oxide synthase (NOS), but how eNOS affects soluble guanylate cyclase (sGC) protein expression in hypoxia-induced pulmonary hypertension is unknown. Wild-type (WT), eNOS-deficient [eNOS(-/-)], and inducible NOS (iNOS)-deficient [iNOS(-/-)] mice were used to investigate the effects of lack of NO from different NOS isoforms on sGC activity and protein expression and its relationship to the muscularization of the pulmonary vasculature. After 6 days of hypoxic exposure (10% O2), the ratios of the right ventricle to left ventricle + septum weight (RV/LV+S) and right ventricle weight to body weight, the lung sGC activity, and vascular muscularization were determined, and protein analysis for eNOS, iNOS, and sGC was performed. Results demonstrated that there were significant increases of RV/LV+S in all animals treated with hypoxia. In hypoxic WT and iNOS(-/-) mice, eNOS and sGC alpha1- and beta1-protein increased twofold; cGMP levels and the number of muscularized vessels also increased compared with hypoxic eNOS(-/-) mice. There was a twofold increase of iNOS protein in WT and eNOS(-/-) mice, and the basal iNOS protein concentration was higher in eNOS(-/-) mice than in WT mice. In contrast, the eNOS(-/-) mouse lung showed no eNOS protein expression, lower cGMP concentrations, and no change of sGC protein levels after hypoxic exposure compared with its normoxic controls (P > 0.34). These results suggest that eNOS, but not iNOS, is a major regulator of sGC activity and protein expression in the pulmonary vasculature. PMID- 11435212 TI - Chronic ethanol ingestion potentiates TNF-alpha-mediated oxidative stress and apoptosis in rat type II cells. AB - In septic patients, chronic alcohol abuse increases the incidence of the acute respiratory distress syndrome (ARDS). Because alveolar type II cell viability is critical for epithelial repair, our objective was to determine if chronic ethanol ingestion increased the sensitivity of type II cells to the inflammatory mediators upregulated during sepsis. In rats chronically fed ethanol, type II cell mitochondrial GSH was depleted, and tumor necrosis factor-alpha (TNF-alpha) induced generation of mitochondrial reactive oxygen species (ROS) and apoptosis were potentiated. When added to the ethanol diet, the GSH precursor (-)-2-oxo-4 thiazolidinecarboxylic acid (Procysteine; Pro) but not N-acetylcysteine (NAC) normalized type II cell mitochondrial GSH. Likewise, Pro but not NAC normalized TNF-alpha-induced mitochondrial ROS and apoptosis. This suggested that chronic ethanol ingestion potentiated TNF-alpha-induced apoptosis in type II cells via mitochondrial GSH depletion. This may be particularly relevant in ARDS when type II cell viability is critical to repair of the damaged alveolar epithelium and may have important ramifications for the treatment of ARDS patients with a history of alcohol abuse. PMID- 11435213 TI - Augmented metalloproteinase activity and acute lung injury in copper-deficient rats. AB - Dietary copper is required for normal function of >30 mammalian enzyme systems. Copper deficiency causes a number of cardiovascular defects as well as impaired immune cell function. Little is known regarding the effects of copper deficiency on acute inflammatory responses, but this topic is relevant because many members of the Western population receive less than the recommended dietary allowance of copper. In the current studies, we investigated the effects of dietary copper deficiency on acute lung injury induced by intrapulmonary deposition of IgG immune complexes. Weanling male Long-Evans rats were fed diets either adequate (5.6 microg/g) or deficient (0.3 microg/g) in copper. IgG immune complex lung injury was greatly increased in copper-deficient rats as determined by lung vascular leakage of albumin and histopathology. However, no change was observed in either the lung content of tumor necrosis factor-alpha or lung neutrophil accumulation. Lungs from copper-deficient rats had much higher levels of matrix metalloproteinase (MMP)-2 and MMP-9 than did copper-adequate control animals. This increased activity was not attributable to alveolar macrophages or neutrophils. These data suggest that the augmented lung injury caused by copper deficiency is due to increased pulmonary MMP-2 and MMP-9 activity and not a generalized amplification of the inflammatory response. PMID- 11435214 TI - Strain dependence of airway hyperresponsiveness reflects differences in eosinophil localization in the lung. AB - Different strains of mice exhibit different degrees of airway hyperresponsiveness after sensitization to and airway challenge with ovalbumin. Antibody responses in BALB/c mice far exceeded those in C57BL/6 mice; in contrast, although responsiveness to methacholine was much higher in the BALB/c mice, the number of eosinophils in the bronchoalveolar lavage fluid was higher in C57BL/6 animals. Sensitized and challenged BALB/c mice developed increases in lung resistance and decreases in dynamic compliance after methacholine or 5-hydroxytryptamine inhalation. C57BL/6 mice only exhibited significant levels of responsiveness when dynamic compliance was monitored in response to inhaled 5-hydroxytryptamine. Eosinophils accumulated in the peribronchial and peripheral lung tissue in BALB/c mice but were distributed diffusely in the peripheral lung tissue of C57BL/6 mice. Thus, in addition to differences in antibody responses and cholinergic agonist reactivity, differences in the responses of large and small airways may reflect the selective distribution of eosinophils in lung tissue. PMID- 11435215 TI - Cigarette smoke extract induces endothelin-1 via protein kinase C in pulmonary artery endothelial cells. AB - We examined the mechanism of endothelin (ET)-1 regulation by cigarette smoke extract (CSE) and the effect of platelets on CSE-induced stimulation of ET-1 gene expression in human and bovine pulmonary artery endothelial cells (PAECs). Our data show that CSE (1%) induces ET-1 gene expression (after 1 h) and ET-1 peptide synthesis (after 4 h) in bovine PAECs. The induction of preproET-1 mRNA level was due to de novo transcription, and new protein synthesis was not required for this induction. The protein kinase C inhibitors staurosporine (10(-8) mol/l) and calphostin C (10(-7) mol/l) abolished the induction of ET-1 gene expression by CSE in bovine and human PAECs. Although a lower concentration of platelets (10(6) cells/ml in bovine PAECs; 10(7) cells/ml in human PAECs) did not significantly alter ET-1 gene expression in PAECs, incubation of platelets with CSE (1%) and PAECs produced a significant increase in preproET-1 mRNA and ET-1 peptide compared with the values in the presence of CSE (1%) alone. CSE (1%) induced platelet aggregation and increased the expression of platelet membrane glycoproteins ex vivo. Thus our data suggest that CSE stimulates ET-1 gene expression via PKC in PAECs. CSE and platelets showed a synergistic effect on ET 1 gene expression, possibly through the activation of platelets by CSE. PMID- 11435216 TI - Human SLPI inactivation after cigarette smoke exposure in a new in vivo model of pulmonary oxidative stress. AB - The role of oxidative stress in inactivating antiproteases is the object of debate. To address this question, we developed an in vivo model of pulmonary oxidative stress induced by cigarette smoke (CS) in mice. The major mouse trypsin inhibitor contrapsin is not sensitive to oxidation, and the mouse secretory leukoprotease inhibitor (SLPI) does not inhibit trypsin. Instead, human recombinant (hr) SLPI inhibits trypsin and is sensitive to oxidation. Thus we determined the effect of CS in vivo on hrSLPI antiproteolytic function in the airways of mice. CS caused a significant decrease in total antioxidant capacity in bronchoalveolar lavage fluid (BALF) and significant changes in oxidized glutathione, ascorbic acid, protein thiols, and 8-epi-PGF(2alpha). Intratracheal hrSLPI significantly increased BALF antitryptic activity. CS induced a 50% drop in the inhibitory activity of hrSLPI. Pretreatment with N-acetylcysteine prevented the CS-induced loss of hrSLPI activity, the decrease in antioxidant defenses, and the elevation of 8-epi-PGF-(2alpha). Thus an inactivation of hrSLPI was demonstrated in this model. This is a novel model for studying in vivo the effects of CS oxidative stress on human protease inhibitors with antitrypsin activity. PMID- 11435217 TI - Interactions between CBP, NF-kappaB, and CREB in the lungs after hemorrhage and endotoxemia. AB - The transcriptional regulatory factor nuclear factor (NF)-kappaB has a central role in modulating expression of proinflammatory mediators that are important in acute lung injury. In vitro studies have shown that competition between NF-kappaB and cAMP response element binding protein (CREB) for binding to the coactivator CREB-binding protein (CBP) is important in regulating transcriptional activity of these factors. In the present study, we examined in vivo interactions between CBP, CREB, and NF-kappaB in hemorrhage- or endotoxemia-induced acute lung injury. Association of CBP with CREB or the p65 subunit of NF-kappaB increased in the lungs after hemorrhage or endotoxemia. Inhibition of xanthine oxidase before hemorrhage, but not before endotoxemia, decreased p65-CBP interactions while increasing those between CREB and CBP. These alterations in CREB-CBP and p65-CBP interactions were functionally significant because xanthine oxidase inhibition before hemorrhage resulted in increased expression of the CREB-dependent gene c Fos and decreased expression of macrophage inflammatory protein-2, a NF-kappaB dependent gene. The present results show that the coactivator CBP has an important role in modulating transcription in vivo under clinically relevant pathophysiological conditions. PMID- 11435218 TI - Migration and gelatinases in cultured fetal, adult, and hyperoxic alveolar epithelial cells. AB - Alveolar epithelial cell (AEC) migration mediated by matrix metalloproteinases (MMPs) is required for lung development and repair after injury such as hyperoxia. Of specific interest in lung remodeling are the gelatinases, which are upregulated in AEC after hyperoxia. We correlated migration and gelatinase production in AEC cultured from fetal, adult, and hyperoxic rats. Fetal AEC (19 20 days) had higher MMP-2 and MMP-9 gelatinase expression than adult AEC, with fivefold higher MMP-9 activity, and were migratory through gelatin, responding to epidermal growth factor, keratinocyte growth factor, and fibroblast growth factor 10. MMP-2 and MMP-9 expression and migratory activity could be detected from the time of plating. In contrast, adult AEC migrated and expressed MMP-2 and MMP-9 proteins only after 48 h of culture. AEC from hyperoxic rats were significantly more migratory through gelatin than control adult AEC, with significantly higher MMP-9 activity. Inhibition of MMPs with doxycycline reduced the migration of AEC from hyperoxic rats to the level of control adult AEC. Fibronectin-cultured "hyperoxic" AEC acquired a temporary capacity for migration similar to the A549 lung cancer cell line, which is both highly migratory and invasive and is derived from the AEC type 2 lineage. These data suggest that MMP activity is associated with a migratory phenotype in fetal, hyperoxic, and transformed AEC in vitro, and we speculate that MMPs may play a key mechanistic role in AEC migration in vivo during lung development and repair. PMID- 11435219 TI - Role of p38 MAP kinase in diperoxovanadate-induced phospholipase D activation in endothelial cells. AB - We previously demonstrated that diperoxovanadate (DPV), a synthetic peroxovanadium compound and cell-permeable oxidant that acts as a protein tyrosine phosphatase inhibitor and insulinomimetic, increased phospholipase D (PLD) activation in endothelial cells (ECs). In this report, the regulation of DPV-induced PLD activation by mitogen-activated protein kinases (MAPKs) was investigated. DPV activated extracellular signal-regulated kinase, c-Jun NH2 terminal kinase (JNK), and p38 MAPK in a dose- and time-dependent fashion. Treatment of ECs with p38 MAPK inhibitors SB-203580 and SB-202190 or transient transfection with a p38 dominant negative mutant mitigated the PLD activation by DPV but not by phorbol ester. SB-202190 blocked DPV-mediated p38 MAPK activity as determined by activated transcription factor-2 phosphorylation. Immunoprecipitation of PLD from EC lysates with PLD1 and PLD2 antibodies revealed both PLD isoforms associated with p38 MAPK. Similarly, PLD1 and PLD2 were detected in p38 immunoprecipitates from control and DPV-challenged ECs. Binding assays demonstrated interaction of glutathione S-transferase-p38 fusion protein with PLD1 and PLD2. Both PLD1 and PLD2 were phosphorylated by p38 MAPK in vitro, and DPV increased phosphorylation of PLD1 and PLD2 in vivo. However, phosphorylation of PLD by p38 failed to affect PLD activity in vitro. These results provide evidence for p38 MAPK-mediated regulation of PLD in ECs. PMID- 11435220 TI - Regulation of anion secretion by nitric oxide in human airway epithelial cells. AB - Nitric oxide (NO) is continuously produced and released in human airways, but the biological significance of this process is unknown. In this study, we have used Calu-3 cells to investigate the effects of NO on transepithelial anion secretion. An inhibitor of NO synthase, NG-nitro-L-arginine methyl ester, reduced short- circuit current (I(sc)), whereas an NO donor, S-nitrosoglutathione (GSNO), increased I(sc), with an EC50 approximately 1.2 microM. The NO-activated current was inhibited by diphenylamine-2-carboxylate, clotrimazole, and charybdotoxin. Selective permeabilization of cell membranes indicated that NO activated both apical anion channels and basolateral potassium channels. An inhibitor of soluble guanylate cyclase, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, prevented activation of I(sc) by NO but not by 8-bromo-cGMP, suggesting that NO acts via a cGMP-dependent pathway. Sequential treatment of cells with forskolin and GSNO or 1-ethyl-2-benzimidazolinone and GSNO showed additive effects of these chemicals on I(sc). Interestingly, GSNO elevated intracellular Ca2+ concentration ([Ca2+]i) but had no effect on I(sc) activated by thapsigargin. These results show that NO activates transepithelial anion secretion via a cGMP-dependent pathway that involves cross talk between NO and [Ca2+]i. PMID- 11435221 TI - Optical method for quantifying rates of mucus secretion from single submucosal glands. AB - We describe an optical method to quantify single- gland secretion. Isolated tracheal mucosa were mounted at the air-Krebs interface and coated with oil. Gland secretions formed spherical bubbles that were digitally imaged at intervals, allowing rates of secretion to be calculated. We monitored 340 glands in 54 experiments with 12 sheep. Glands secreted basally at low rates (0.57 +/- 0.04 nl x min(-1) x gland(-1), 123 glands) in tissues up to 9 h postharvest and at lower rates for up to 3 days. Carbachol (10 microM) stimulated secretion with an early transient and a sustained or oscillating phase. Peak secretion was 15.7 +/- 1.2 nl x min(-1) x gland(-1) (60 glands); sustained secretion was 4.5 +/- 0.5 nl x min(-1) x gland(-1) (10 glands). Isoproterenol and phenylephrine (10 microM each) stimulated only small, transient responses. We confirmed that cats have a large secretory response to phenylephrine (11.6 +/- 3.7 nl x min(-1) x gland(-1), 12 glands), but pigs, sheep, and humans all have small responses (<2 nl x min( 1)m x gland(-1)). Carbachol-stimulated peak secretion was inhibited 56% by bumetanide, 67% by HCO replacement with HEPES, and 92% by both. The distribution of secretion rates was nonnormal, suggesting the existence of subpopulations of glands. PMID- 11435222 TI - EP2 receptors mediate airway relaxation to substance P, ATP, and PGE2. AB - Substance P (SP) and ATP evoke transient, epithelium-dependent relaxation of constricted mouse tracheal smooth muscle. Relaxation to either SP or ATP is blocked by indomethacin, but the specific eicosanoid(s) involved have not been definitively identified. SP and ATP are reported to release PGE2 from airway epithelium in other species, suggesting PGE2 as a likely mediator in epithelium dependent airway relaxation. Using mice homozygous for a gene-targeted deletion of the EP2 receptor [EP2(-/-)], one of the PGE2 receptors, we tested the hypothesis that PGE2 is the primary mediator of relaxation to SP or ATP. Relaxation in response to SP or ATP was significantly reduced in tracheas from EP2(-/-) mice. There were no differences between EP2(-/-) and wild-type tracheas in their physical dimensions, contraction to ACh, or relaxation to isoproterenol, thus ruling out any general alterations of smooth muscle function. There were also no differences between EP2(-/-) and wild-type tracheas in basal or stimulated PGE2 production. Exogenous PGE2 produced significantly less relaxation in EP2(-/-) tracheas compared with the wild type. Taken together, this experimental evidence supports the following two conclusions: EP2 receptors are of primary importance in airway relaxation to PGE2 and relaxation to SP or ATP is mediated through PGE2 acting on EP2 receptors. PMID- 11435223 TI - eNOS and prostanoid enzymes in lungs of newborn piglets with chronic aortopulmonary shunts. AB - Our purpose was to determine if abundance of proteins underlying nitric oxide (NO) and prostanoid production is altered in lungs of piglets with aortopulmonary shunts. We also evaluated whether shunted piglets exhibit abnormal pulmonary vascular responses to ACh, an endothelium-dependent agent that mediates dilation in part by NO and prostanoid release. At age 4-5 days, piglets underwent either a sham operation or placement of an aortopulmonary shunt. At age 5-6 wk, pulmonary arterial pressure (Ppa) and cardiac output by the thermodilution technique were measured in anesthetized piglets. Ppa responses to the endothelium-dependent agent, ACh, and to a non-endothelium-dependent agent, papaverine, were measured in perfused lungs. An immunoblot technique was applied to homogenates of whole lung tissue and two size groups of pulmonary arteries. In shunted piglets, Ppa and cardiac output were elevated, and Ppa responses to papaverine were reduced. ACh responses were not decreased when expressed relative to Ppa dilation with papaverine. Endothelial nitric oxide synthase (eNOS), cyclooxygenase-1, cyclooxygenase-2, prostacyclin synthase, and thromboxane synthase amounts were unaltered in all lung tissue homogenates. Altered abundance of eNOS and/or prostanoid enzymes does not contribute to the blunted dilation and the elevation in Ppa associated with aortopulmonary shunts in newborn piglets. PMID- 11435224 TI - Activation of the EGF receptor signaling pathway in airway epithelial cells exposed to Utah Valley PM. AB - Exposure to ambient particulate matter (PM) in the Utah Valley has previously been associated with a variety of adverse health effects. To investigate intracellular signaling mechanisms for pulmonary responses to Utah Valley PM inhalation, human primary airway epithelial cells were exposed to aqueous extracts of PM collected from the year before (Y1), during (Y2), and after (Y3) the closure of a local steel mill located in the Utah Valley in this study. Transfection with kinase-deficient extracellular signal-regulated kinase (ERK) 1 constructs partially blocked Utah Valley PM-induced interleukin (IL)-8 promoter reporter activity. The mitogen-activated protein kinase/ERK kinase (MEK) activity inhibitor PD-98059 significantly abolished IL-8 released in response to Utah Valley PM, as did the epidermal growth factor (EGF) receptor kinase inhibitor AG 1478. Western blotting showed that Utah Valley PM induced phosphorylation of EGF receptor tyrosine, MEK1/2, and ERK1/2, which could be ablated with AG-1478 or PD 98059. For all findings, the potency of Utah Valley PM collected during Y2 was found to be lower relative to that of Y1 and Y3. These data demonstrate that Utah Valley PM can induce IL-8 expression partially through the activation of the EGF receptor signaling. PMID- 11435225 TI - Shear stress regulation of endothelial NOS in fetal pulmonary arterial endothelial cells involves PKC. AB - We have shown that increased pulmonary blood flow at birth increases the activity and expression of endothelial nitric oxide (NO) synthase (eNOS). However, the signal transduction pathway regulating this process is unclear. Because protein kinase C (PKC) has been shown to be activated in response to shear stress, we undertook a study to examine its role in mediating shear stress effects on eNOS. Initial experiments demonstrated that PKC activity increased in response to shear stress. NO production in response to shear stress was found to be biphasic, with an increase in NO release up to 1 h, a plateau phase until 4 h, and another increase between 4 and 8 h. PKC inhibition reduced the initial rise in NO release by 50% and the second increase by 70%. eNOS mRNA and protein levels were also increased in response to shear stress, whereas PKC inhibition prevented this increase. The stimulation of PKC activity with phorbol ester increased eNOS gene expression without increasing NO release. These results suggest that PKC may play different roles in shear stress-mediated release of NO and increased eNOS gene expression. PMID- 11435226 TI - Gene suppression by tristetraprolin and release by the p38 pathway. AB - Tristetraprolin (TTP) is a zinc finger protein that has been implicated in the control of tumor necrosis factor (TNF) mRNA stability. We show here that TTP protein has a suppressive effect on promoter elements from TNF-alpha and interleukin-8 and that lipopolysaccharide (LPS) stimulation can release this suppression. The release in LPS-stimulated cells was found to be primarily mediated by the p38 pathway because activation of p38 is sufficient to remove the suppressive effect of TTP. Indeed, TTP seems to be a direct substrate of p38 in vivo since it is an excellent substrate of p38 in vitro, and mutation of potential phosphorylation sites in TTP prevents release of the suppression imposed on TNF transcription. We found TTP protein to be present at low levels in the resting macrophage cell line RAW 264.7 and to be quickly induced after LPS stimulation. The kinetics of TTP induction suggests a potential role of TTP as an important player in switching off LPS-induced genes after induction. In conclusion, TTP plays an important role in maintaining gene quiescence, and this quenching effect on transcription can be released by p38 phosphorylation of TTP. PMID- 11435227 TI - Cytotoxic effects of cigarette smoke extract on an alveolar type II cell-derived cell line. AB - Injury of the alveolar epithelium by cigarette smoke is presumed to be an important process in the pathogenesis of smoking-related pulmonary diseases. We investigated the cytotoxic effects of cigarette smoke extract (CSE) on an alveolar type II cell-derived cell line (A549). CSE caused apoptosis at concentrations of 5% or less and necrosis at 10% or more. When CSE was exposed to air before application to A549 cells, the cytotoxic effects were attenuated. CSE caused cell death without direct contact with the cells. Acrolein and hydrogen peroxide, two major volatile factors in cigarette smoke, caused cell death in a similar manner. Aldehyde dehydrogenase, a scavenger of aldehydes, and N acetylcysteine, a scavenger of oxidants and aldehydes, completely inhibited CSE induced apoptosis. CSE and acrolein increased intracellular oxidant activity. In conclusion, apoptosis of alveolar epithelial cells may be one of the mechanisms of lung injury induced by cigarette smoking. This cytotoxic effect might be due to an interaction between aldehydes and oxidants present in CSE or formed in CSE exposed cells. PMID- 11435228 TI - Perfluorocarbon enhanced gas exchange: the easy way. PMID- 11435229 TI - Jails, chest radiography, and the elimination of tuberculosis. PMID- 11435230 TI - Bronchial eosinophilia in exacerbation of bronchitis: an allergic profile of inflammation? PMID- 11435231 TI - On the merit of making comparisons. PMID- 11435232 TI - Treatment of HIV-related tuberculosis in the era of effective antiretroviral therapy. PMID- 11435233 TI - Treating nicotine addiction: not a medical problem? PMID- 11435234 TI - Sleep-disordered breathing in children. PMID- 11435235 TI - Persistent improvement of gas exchange and lung mechanics by aerosolized perfluorocarbon. AB - The effect of aerosolized perfluorocarbon (PFC) (FC77) on pulmonary gas exchange and lung mechanics was studied in a surfactant depleted piglet model. Sixty minutes after induction of lung injury by bronchoalveolar lavage, 20 piglets were randomized to receive aerosolized PFC (Aerosol-PFC, 10 ml/kg/h, n = 5), partial liquid ventilation (PLV) at FRC capacity volume (FRC-PLV, 30 ml/kg, n = 5) or low volume (LV-PLV, 10 ml/kg/h, n = 5), or intermittent mandatory ventilation (IMV) (Control, n = 5). After 2 h, perfluorocarbon application was stopped and IMV was continued for 6 h. Sixty minutes after the onset of therapy, PaO2 was significantly higher and PaCO2 was significantly lower in the Aerosol-PFC and the FRC-PLV groups than in the LV-PLV and the Control groups; p < 0.001. Six hours after treatment, maximum PaO2 was found in the Aerosol-PFC group: 406.4 +/- 26.9 mm Hg, FRC-PLV: 217.3 +/- 50.5 mm Hg, LV-PLV: 96.3 +/- 18.9 mm Hg, Control: 67.6 +/- 8.4 mm Hg; p < 0.001. PaCO2 was lowest in the Aerosol-PFC group: 24.2 +/- 1.7 mm Hg, FRC-PLV: 35.9 +/- 2.8 mm Hg, LV-PLV: 56.7 +/- 12.4 mm Hg, Control: 60.6 +/ 5.1 mm Hg; p < 0.01. Dynamic compliance (C20/c) was highest in the Aerosol-PFC group; p < 0.01. Aerosolized perfluorocarbon improved pulmonary gas exchange and lung mechanics as effectively as PLV did in surfactant-depleted piglets, and the improvement was sustained longer. PMID- 11435236 TI - Preserved spontaneous breathing improves cardiac output during partial liquid ventilation. AB - The aim of this study was to examine whether preserved spontaneous breathing (SB) supported by proportional-assist ventilation (PAV) would improve cardiac output (CO) during partial liquid ventilation (PLV) in rabbits with and without lung disease if compared with time-cycled, volume-controlled ventilation (CV) combined with muscle paralysis (MP). PLV was initiated in 17 healthy rabbits and 17 surfactant-depleted rabbits using 12 to 15 ml/kg of perfluorodecaline. Both ventilatory modes, SB+PAV and CV+MP, were applied in random sequence using a crossover design. CO was measured by thermodilution. CO was significantly higher during SB+PAV than during CV+MP: 136 +/- 21 ml/kg x min (mean +/- SD) versus 120 +/- 30 ml/kg x min (p = 0.004) in healthy rabbits, and 147 +/- 19 ml/kg x min versus 111 +/- 13 ml/kg x min (p < 0.0001) in surfactant-depleted rabbits, resulting in an improved oxygen delivery. This difference was mainly caused by a larger stroke volume during SB+PAV, whereas there was little change in heart rate. In surfactant-depleted rabbits, SB+PAV resulted in improved arterial blood pressure and arterial and mixed venous pH and in a higher PaO2 at the same level of PEEP and mean airway pressure. We conclude that during PLV, CO is higher during SB+PAV than during CV+MP, resulting in an improved oxygen delivery. In surfactant-depleted rabbits, improved CO, oxygen delivery, and arterial blood pressure resulted in higher pH, possibly reflecting improved tissue perfusion and oxygenation. PMID- 11435237 TI - Long-term effects of spontaneous breathing during ventilatory support in patients with acute lung injury. AB - Improved gas exchange has been observed during spontaneous breathing with airway pressure release ventilation (APRV) as compared with controlled mechanical ventilation. This study was designed to determine whether use of APRV with spontaneous breathing as a primary ventilatory support modality better prevents deterioration of cardiopulmonary function than does initial controlled mechanical ventilation in patients at risk for acute respiratory distress syndrome (ARDS). Thirty patients with multiple trauma were randomly assigned to either breathe spontaneously with APRV (APRV Group) (n = 15) or to receive pressure-controlled, time-cycled mechanical ventilation (PCV) for 72 h followed by weaning with APRV (PCV Group) (n = 15). Patients maintained spontaneous breathing during APRV with continuous infusion of sufentanil and midazolam (Ramsay sedation score [RSS] of 3). Absence of spontaneous breathing (PCV Group) was induced with sufentanil and midazolam (RSS of 5) and neuromuscular blockade. Primary use of APRV was associated with increases (p < 0.05) in respiratory system compliance (CRS), arterial oxygen tension (PaO2), cardiac index (CI), and oxygen delivery (DO2), and with reductions (p < 0.05) in venous admixture (QVA/QT), and oxygen extraction. In contrast, patients who received 72 h of PCV had lower CRS, PaO2, CI, DO2, and Q VA/Q T values (p < 0.05) and required higher doses of sufentanil (p < 0.05), midazolam (p < 0.05), noradrenalin (p < 0.05), and dobutamine (p < 0.05). CRS, PaO2), CI and DO2 were lowest (p < 0.05) and Q VA/Q T was highest (p < 0.05) during PCV. Primary use of APRV was consistently associated with a shorter duration of ventilatory support (APRV Group: 15 +/- 2 d [mean +/- SEM]; PCV Group: 21 +/- 2 d) (p < 0.05) and length of intensive care unit (ICU) stay (APRV Group: 23 +/- 2 d; PCV Group: 30 +/- 2 d) (p < 0.05). These findings indicate that maintaining spontaneous breathing during APRV requires less sedation and improves cardiopulmonary function, presumably by recruiting nonventilated lung units, requiring a shorter duration of ventilatory support and ICU stay. PMID- 11435238 TI - A method for measuring passive elastance during proportional assist ventilation. AB - There are currently no reliable, noninvasive ways to monitor respiratory elastance (E) during assisted ventilation. We describe a method that is suited for proportional assist ventilation (PAV). In this mode, the end of the ventilator's inflation phase occurs during the declining phase of inspiratory effort (Pmus). If the opening of the exhalation valve is delayed, airway pressure (Paw) should initially rise as Pmus continues its decline. When Pmus declines to zero, a Paw plateau should appear. Paw at this point should reflect passive recoil at the prevailing volume. A cohort of 74 ventilator-dependent patients, ventilated in the PAV mode, were studied. Brief end-inspiratory occlusions were applied at random intervals. The magnitude of early change in Paw during the occlusion was inversely related to level of assist (r = 0.7, p < 0.00001). At high assist (> 75%), Paw was nearly flat or declined slightly, indicating minimal residual Pmus at the onset of occlusion. At lower assist levels, Paw increased exponentially in most patients with an average time constant of 0.21 +/- 0.06 s. Extraneous events that may corrupt the measurement (e.g., behavioral responses) were extremely rare (< 0.5%) in the first 0.25 s. From these findings, we concluded that Paw measured 0.25 s from occlusion onset (P0.25) includes little inspiratory Pmus and is free of extraneous events. E, estimated from P0.25 during PAV (EPAV), agreed well (r = 0.92) with passive E measured during controlled ventilation (ECMV); the average difference (EPAV - ECMV) was (+/- SD) -0.3 +/- 4.9 cm H2O x L(-1), corresponding to 0.9 +/- 16.4% of average E. We conclude that Paw measured at 0.25 s from the onset of end-inspiratory occlusion in the PAV mode provides a reliable estimate of passive elastic recoil. PMID- 11435239 TI - Chronic obstructive pulmonary disease and intake of catechins, flavonols, and flavones: the MORGEN Study. AB - Flavonoids have been suggested to protect against chronic lung disease. We studied intake of catechins, flavonols, and flavones in relation to pulmonary function and COPD symptoms in 13,651 adults from three Dutch cities examined from 1994 to 1997. Dietary intake was estimated using a food frequency questionnaire, and flavonoid intake was calculated using specific food composition tables. Pulmonary function (FEV1) was determined by spirometry and COPD symptoms by questionnaire. Associations were presented for the fifth versus the first quintile of intake (Q5-Q1), adjusted for age, height (for FEV1 only), sex, smoking, BMI, and energy intake. Smoking was strongly associated with COPD, independent of dietary effects. Average catechin, flavonol, and flavone intake was 58 mg/d (SD = 46) with tea and apples as main sources. Total catechin, flavonol, and flavone intake was positively associated with FEV1 (beta(Q5-Q1) = 44 ml, 95% CI = 18-69) and inversely associated with chronic cough (OR(Q5-Q1) = 0.80, 95% CI = 0.66-0.97) and breathlessness (OR(Q5-Q1) = 0.74, 95% CI = 0.58- 0.94), but not chronic phlegm. Catechin intake was independently associated with FEV1 (beta(Q5-Q1) = 130 ml, 95% CI = 101-159) and all three COPD symptoms (OR(Q5 Q1) = 0.60-0.72, p < 0.001). Flavonol and flavone intake was independently associated with chronic cough only. Solid fruit, but not tea, intake was beneficially associated with COPD. Our results suggest a beneficial effect of a high intake of catechins and solid fruits against COPD. PMID- 11435240 TI - Current indoor allergen levels of fungi and cats, but not house dust mites, influence allergy and asthma in adults with high dust mite exposure. AB - We assessed the influence of current indoor levels of fungi, house dust mite allergen (Der p 1), and cat allergen (Fel d 1) on sensitization and asthma in adults. A total of 485 adults answered a questionnaire and had skin prick tests and lung function tests. Dust and air samples were collected from their bedrooms. The dust was analyzed for Der p 1, Fel d 1, and fungal biomass (ergosterol). Fungal propagules were measured in air samples. Current asthma was defined as having wheezed during the past 12 mo plus bronchial hyperreactivity (BHR) to methacholine. High exposure to total airborne fungi was associated with increased BHR, but perhaps paradoxically with a lower risk of being sensitized to fungi. Ergosterol levels in floor dust were a risk factor both for being sensitized to fungi and having wheezed within the last year. High Fel d 1 levels in floor dust were found to increase the risk of being sensitized to cats and in beds to increase the risk of current asthma. Although Der p 1 levels in homes were high, people exposed to high Der p 1 levels in floor dust were less likely to be sensitized to house dust mites or to have wheezed within the past year. Current indoor levels of fungi and Fel d 1, but not Der p 1, influenced sensitization and asthma in adults with high dust mite exposure. PMID- 11435241 TI - Prevalence of upper respiratory symptoms in patients with symptomatic gastroesophageal reflux disease. AB - This study evaluated the prevalence of upper respiratory symptoms (URS) among patients with symptomatic gastroesophageal reflux disease (GERD). Seventy-four subjects with heartburn completed a URS questionnaire before dual-probe, 24-h esophageal pH monitoring. The URS questionnaire was also completed by 74 normal volunteers without previous or current symptoms of GERD. Esophageal pH monitoring results were classified as normal, distal, or proximal and distal gastroesophageal reflux using standardized criteria. Mean URS scores (+/- SD) were 8.31 +/- 3.98 in the 52 subjects with GERD and 4.57 +/- 3.57 in the 22 subjects with negative pH probe studies, p = 0.02. Subjects with negative pH probe studies and normal volunteers scored similarly on the URS questionnaire. Reflux episodes/24 h correlated with URS scores, r = 0.47, p = 0.0001. Seventy five percent of subjects with upper reflux, 68% of subjects with lower reflux, 36% of subjects with normal esophageal pH studies, and 9% of normal volunteers reported laryngeal symptoms for at least 5 d/mo. Sixty-nine percent of subjects with upper reflux, 50% of subjects with lower reflux, 31% of subjects with normal pH studies, and 14% of normal volunteers reported nasal symptoms for at least 5 d/mo. URS are frequent among subjects with GERD. KEYWORDS: rhinitis; upper airway; gastroesophageal reflux PMID- 11435242 TI - Miniature chest radiograph screening for tuberculosis in jails: a cost effectiveness analysis. AB - Jails are an important reservoir of tuberculosis infection in the United States. Screening for active disease in these high-risk settings is difficult. We used decision analysis to assess the cost effectiveness of routine miniature chest radiography for screening for tuberculosis on admission to jail. Infection rates, probabilities, and costs associated with detecting and treating tuberculosis were derived from published studies. We calculated an average total cost of $6.60 per inmate for routine radiograph screening on admission to jail. The cost of screening for active tuberculosis with miniature chest radiography was estimated to be $9,600 per case identified, compared with $32,100 per case with tuberculin skin testing and $54,100 per case with symptom screening. Chest radiography would also identify substantially more cases than other methods of screening. Screening for tuberculosis with miniature chest radiography is cost effective even under a wide range of assumptions regarding risk factors and prevalence of disease. Miniature chest radiography should be strongly considered as an important tool in the fight to eliminate tuberculosis from the high-risk populations that may be reached through screening in jails. PMID- 11435243 TI - Use of audio computer-assisted self-interviews to assess tuberculosis-related risk behaviors. AB - The objective of this study was to compare self-reported tuberculosis and human immunodeficiency virus (HIV) risk factors obtained from computer-assisted questionnaires and interviewer-assisted questionnaires among participants of a needle exchange program. Between June 1998 and May 1999, needle exchange program participants requesting tuberculosis screening underwent interviews regarding demographics and risk factors for tuberculosis and HIV infection. The first 190 participants underwent traditional interviewer-assisted questionnaires, whereas the remaining 92 underwent computer-assisted questionnaires. Data were analyzed by interview technique using odds ratios (OR) and multiple logistic regression. Among 282 participants, demographic characteristics, health status, HIV serostatus, visits to homeless shelters, alcohol intake, and cigarette smoking were all similar by interview technique. However, respondents receiving computer assisted questionnaires were more likely than those receiving interviewer assisted questionnaires to report smoking marijuana (OR = 5.56), crack (OR = 1.88), and heroin (OR = 2.60); as well as sharing cocaine smoking equipment (OR = 4.49), sharing heroin smoking equipment (OR = 2.85), "shotgunning" (OR = 4.48), and visiting crack houses (OR = 4.39). In the final multivariate model, respondents receiving computer-assisted questionnaires were more likely to report "shotgunning" and visiting a crack house relative to respondents receiving interviewer-assisted questionnaires. In conclusion, increased odds of high-risk behaviors for tuberculosis and HIV infection among computer-assisted questionnaire respondents support the use of computer-assisted questionnaires to ascertain risk behavior data for both tuberculosis and HIV. KEYWORDS: tuberculosis; HIV; self-report; drug use; computer-assisted PMID- 11435244 TI - The effect of nebulized epinephrine on respiratory mechanics and gas exchange in bronchiolitis. AB - Nebulized epinephrine has been advocated as a treatment for airway obstruction in infants with bronchiolitis; however, its effect on respiratory mechanics and gas exchange has been poorly documented to date. We performed a preinterventional and postinterventional study with primary outcome measures of mechanics (measured by single-breath occlusion passive deflation) and oxygenation and ventilation indices in order to measure the effects of nebulized epinephrine in infants requiring mechanical ventilation for RSV-positive bronchiolitis. A two compartment model was used to describe respiratory mechanics in patients with nonlinear flow-volume curves. Nebulized epinephrine (0.5 mg/kg) was administered to 15 patients (median age, 0.19 yr; weight, 4.4 kg). Resistance decreased significantly in slow and fast compartments in 87 and 70% of patients, respectively. Median resistance in the slow compartment decreased from 0.427 to 0.198 cm H2O/ml/s (p = 0.0015) and in the fast compartment from 0.167 to 0.116 cm H2O/ ml/s (p = 0.018). Compliance, oxygenation index, and ventilation index were not significantly changed after administration of epinephrine. We conclude that nebulized epinephrine substantially improves respiratory system resistance but not oxygenation or ventilation indices. This may be because of the effects of epinephrine on oxygen consumption or ventilation-perfusion matching. PMID- 11435245 TI - Disproportionate distribution of Burkholderia cepacia complex species and transmissibility markers in cystic fibrosis. AB - Several distinct species (genomovars) comprise bacteria previously identified merely as Burkholderia cepacia. Understanding how these species, collectively referred to as the B. cepacia complex, differ in their epidemiology and pathogenic potential in cystic fibrosis (CF) is important in efforts to refine management strategies. B. cepacia isolates recovered from 606 CF patients receiving care at 132 treatment centers in 105 cities in the United States were assessed to determine species within the B. cepacia complex and examined for the presence of putative transmissibility markers (B. cepacia epidemic strain marker [BCESM] and cable pilin subunit gene [cblA]). Fifty percent of patients were infected with B. cepacia complex genomovar III, 38% with B. multivorans (formerly genomovar II), and 5% with B. vietnamiensis (formerly genomovar V); fewer than 5% of patients were infected with either genomovar I, B. stabilis (formerly genomovar IV), genomovar VI, or genomovar VII. BCESM was found in 46% of genomovar III isolates and not in any other species. Only one isolate, from a patient infected with the ET12 epidemic lineage, contained the complete cblA pilin subunit gene. Our data indicate a differential capacity for human infection among the phylogenetically closely related species of the B. cepacia complex. The low frequency of BCESM and cblA suggests that they are not sufficient markers of B. cepacia virulence or transmissibility. PMID- 11435246 TI - Neutrophils, unopposed neutrophil elastase, and alpha1-antiprotease defenses following human lung transplantation. AB - Neutrophils are sequestered in the newly transplanted lung after reperfusion or with infection, rejection, and chronic graft dysfunction. Because unopposed (free) neutrophil elastase (NE) released into bronchoalveolar secretions may injure the lung allograft and impair bacterial clearance, we assessed total neutrophil numbers, myeloperoxidase activity as an index of neutrophil influx and degranulation, alpha1-antiprotease (alpha1-AP) concentrations, and unopposed NE activity in bronchoalveolar secretions from lung transplant recipients. Unopposed NE activity was present in bronchoalveolar lavage fluid (BALF) from recipients transplanted for emphysema associated with alpha1-AP deficiency as well as recipients without such deficiency (171 of 2,137 BALF; 8%). Ten of 17 (59%) recipients with alpha1-AP deficiency who were followed for at least 1 yr after transplant with multiple surveillance and diagnostic bronchoscopies had at least one BALF containing unopposed NE, usually associated with the presence of > or = 10(5) colony forming units/ml BALF of aerobic bacteria. In contrast, 19 of 58 (33%) with emphysema not associated with alpha1-AP deficiency, 8 of 32 (25%) recipients with cystic fibrosis (CF), 6 of 16 (38%) with idiopathic pulmonary fibrosis (IPF), and 11 of 36 (31%) with other indications for transplant had unopposed NE in BALF. alpha1-AP levels were significantly elevated in the early posttransplant time period and could be augmented considerably in alpha1-AP deficient recipients with episodes of infection or rejection. Our findings indicate that unopposed NE activity can be found in both alpha1-AP-deficient and alpha1-AP-sufficient recipients after transplantation, usually in association with endobronchial bacterial infection. PMID- 11435247 TI - Pulmonary function in idiopathic pulmonary fibrosis and referral for lung transplantation. AB - Of patients awaiting lung transplantation, the death rates are highest in those with idiopathic pulmonary fibrosis (IPF), suggesting that many IPF patients are referred late for transplantation. Therefore this study was undertaken to evaluate baseline pulmonary function test (PFT) and high-resolution computed tomography (HRCT) fibrosis scores, and the relationship to survival in IPF patients younger than 65 yr of age. A total of 115 patients with usual interstitial pneumonia (UIP) were studied. At presentation to a tertiary referral center, PFT and HRCT data were collected and analyzed for prognostic significance: the primary outcome measure was patient death. Based on the length of the waiting list for transplantation, prediction of 2-yr survival was examined. DL(CO) percent predicted and HRCT-fibrosis score were found to be independent predictors of survival and in combination gave the best prognostic prediction. The optimal points on the receiver operating characteristic (ROC) curves for discriminating between survivors and nonsurvivors corresponded to 39% DL(CO) percent predicted, and to a HRCT-fibrosis score of 2.25. The combination of these parameters yielded an optimal point with a specificity and a sensitivity of 84% and 82%, respectively. A model based on a combination of DL(CO) percent predicted and HRCT-fibrosis score may optimize the timing of referral for transplantation. PMID- 11435248 TI - Exacerbations of Bronchitis: bronchial eosinophilia and gene expression for interleukin-4, interleukin-5, and eosinophil chemoattractants. AB - Eosinophilia has been reported during exacerbations of bronchitis, but the mechanisms of tissue recruitment of eosinophils are unclear. We quantified eosinophils and the concurrent expression of cytokines and chemokines probably responsible for the tissue eosinophilia in bronchial biopsies obtained from three groups of nonatopic subjects: (1) healthy nonsmokers (n = 7; FEV1 % predicted = 108 +/- 4 [mean +/- SEM]); (2) nonasthmatic smokers with chronic bronchitis (CB) in a stable phase of their disease (n = 11; FEV1 % predicted: 75 +/- 5); and (3) nonasthmatic subjects with CB who sought medical advice for an exacerbation of their condition (n = 9; FEV(1) % predicted: 61 +/- 8). We applied anti-EG2 antibody and immunostaining to detect and count eosinophils. We performed in situ hybridization to visualize and enumerate cells expressing the genes for interleukin (IL)-4 and IL-5 and the eosinophil chemokines eotaxin, monocyte chemoattractant protein (MCP)-4, or regulated on activation, normal T-cell expressed and secreted (RANTES). We confirmed an increase in EG2-positive eosinophils in patients with CB in exacerbation. We found messenger RNA (mRNA) positivity for IL-4 and IL-5 in CB, but the between-group differences were not statistically significant. However, the numbers of lymphomononuclear cells expressing eotaxin mRNA were significantly greater in the smokers with CB than in the healthy nonsmokers without CB (p < 0.01). Following an exacerbation, RANTES expression was upregulated and this chemokine was strongly expressed in both the surface epithelium and in subepithelial lymphomononuclear cells: only RANTES showed a significant positive correlation with the increasing number of EG2 positive cells (r = 0.51; p < 0.03). In conclusion, an allergic profile of inflammation can also occur in CB: the marked upregulation of RANTES in the epithelium and subepithelium most likely accounts for the increased eosinophilia associated with an exacerbation of bronchitis. PMID- 11435249 TI - Effect of a long-acting beta2-agonist over three months on airway wall vascular remodeling in asthma. AB - There are few data regarding the potential effects of antiasthma treatment on indices of airway remodeling, such as the increased subepithelial airway vascularity in patients with asthma. We studied 45 symptomatic subjects with asthma who were receiving treatment with low dose inhaled corticosteroids (ICS) (range 200-500 microg twice a day) and 28 normal subjects without asthma as a control population. Subjects underwent bronchoscopy with airway biopsy and subjects with asthma were then randomized to receive supplementary inhaled salmeterol 50 microg twice a day, fluticasone propionate 100 microg twice a day, or placebo for 3 mo in addition to their baseline ICS. Biopsy of the airway was then repeated. The biopsies were analyzed for vascular structures in the subepithelial lamina propria. Sufficient biopsy material was available for analysis of vascularity in 34 of the subjects with asthma and 28 of the normal subjects. We confirmed that airways of subjects with asthma had a significant increase in the number of vessels/mm2 of lamina propria compared with airways of normal subjects (524 +/- 137 vessels/mm2, n = 34 versus 425 +/- 130 vessels/mm2, n = 28; p = 0.004). There was a decrease in the density of vessels of lamina propria after treatment only in the salmeterol group compared with baseline (before, 535 +/- 153 vessels/mm2 versus after, 400 +/- 142 vessels/mm2; n = 12; p = 0.04). There was no significant change within the fluticasone (n = 11) or placebo (n = 11) treatment groups, but also no significant differences between the groups. Notably, no treatment was associated with increased airway wall vascularity. The demonstrated fall in vessel number within the salmeterol-treated group may suggest an advantageous effect of long-acting beta2-agonists on this manifestation of airway remodeling over the 3-mo time scale of this study, which is complementary to the action of ICS on airway vascularity. PMID- 11435250 TI - Recruitment and derecruitment during acute respiratory failure: an experimental study. AB - We aimed to elucidate the relationships between pleural (Ppl), esophageal (Pes), and superimposed gravitational pressures in acute lung injury, and to understand the mechanisms of recruitment and derecruitment. In six dogs with oleic acid respiratory failure, we measured Pes and Ppl in the uppermost, middle, and most dependent lung regions. Each dog was studied at positive end-expiratory pressure (PEEP) of 5 and 15 cm H2O and three levels of tidal volume (VT; low, medium, and high). For each PEEP-VT combination, we obtained a computed tomographic (CT) scan at end-inspiration and end-expiration. The variations of Ppl and Pes pressures were correlated (r = 0.86 +/- 0.07, p < 0.0001), as was the vertical gradient of transpulmonary (PL) and superimposed pressure (r = 0.92, p < 0.0001). Recruitment proceeded continuously along the entire volume-pressure curve. Estimated threshold opening pressures were normally distributed (mode = 20 to 25 cm H2O). The amount of end-expiratory collapse at the same PEEP and PL was significantly lower when ventilation was performed at high VT. End-inspiratory and end expiratory collapse were highly correlated (r = 0.86, p < 0.0001), suggesting that as more tissue is recruited at end-inspiration, more remains recruited at end-expiration. When superimposed pressure exceeded applied airway pressure (Paw), collapse significantly increased. PMID- 11435251 TI - Recruitment and derecruitment during acute respiratory failure: a clinical study. AB - In a model of acute lung injury, we showed that positive end-expiratory pressure (PEEP) and tidal volume (VT) are interactive variables that determine the extent of lung recruitment, that recruitment occurs across the entire range of total lung capacity, and that superimposed pressure is a key determinant of lung collapse. Aiming to verify if the same rules apply in a clinical setting, we randomly ventilated five ALI/ARDS patients with 10, 15, 20, 30, 35, and 45 cm H2O plateau pressure and 5, 10, 15, and 20 cm H2O of PEEP. For each PEEP-VT condition, we obtained computed tomography at end inspiration and end expiration. We found that recruitment occurred along the entire volume-pressure curve, independent of lower and upper inflection points, and that estimated threshold opening pressures were normally distributed (mode = 20 cm H2O). Recruitment occurred progressively from nondependent to dependent lung regions. Overstretching was not associated with hyperinflation. Derecruitment did not parallel deflation, and estimated threshold closing pressures were normally distributed (mode = 5 cm H2O). End-inspiratory and end-expiratory collapse were correlated, suggesting a plateau-PEEP interaction. When superimposed gravitational pressure exceeded PEEP, end-expiratory collapse increased. We concluded that the rules governing recruitment and derecruitment equally apply in an oleic acid model and in human ALI/ARDS. PMID- 11435252 TI - Direct effects of interleukin-13 on signaling pathways for physiological responses in cultured human airway smooth muscle cells. AB - Numerous studies have suggested an important role for the Th2 cytokines interleukin (IL)-13 and IL-4 in the development of allergic asthma. We tested the hypothesis that IL-13 and IL-4 have direct effects on cultured airway smooth muscle cells (HASM). Using RT-PCR, we showed that HASM cells express transcripts for IL-4alpha, IL-13RalphaI, and IL-13RalphaII, but not for the common IL-2Rgamma chain. We then analyzed the capacity of the two cytokines to activate signaling pathways in HASM cells. Both IL-13 and IL-4 caused STAT-6 phosphorylation, but the time course was different between the two cytokines, with peak effects occurring 15 min after addition of IL-4 and 1 h after addition of IL-13. Effects on signaling were observed at cytokine concentrations as low as 0.3 ng/ml. IL-4 and IL-13 also caused phosphorylation of ERK MAP kinase. As suggested by the signaling studies, the biological responses of the two cytokines were also different. We used magnetic twisting cytometry to measure cell stiffness of HASM cells and tested the capacity of IL-4 and IL-13 to interfere with the reductions in cell stiffness induced by the beta-agonist isoproterenol (ISO). IL-13 (50 ng/ml for 24 h), but not IL-4, significantly reduced beta-adrenergic responsiveness of HASM cells, and the MEK inhibitor U0126 significantly reduced the effects of IL-13 on ISO-induced changes in cell stiffness. We propose that these direct effect of IL-13 on HASM cells may contribute at least in part to the airway narrowing observed in patients with asthma. PMID- 11435253 TI - Decreased pulmonary and tracheal smooth muscle expression and activity of type 1 nitric oxide synthase (nNOS) after ovalbumin immunization and multiple aerosol challenge in guinea pigs. AB - Pharmacological evidence supports a role of a transient decreased endogenous nitric oxide (NO) synthesis in ovalbumin (OVA)-induced early airway hyperresponsiveness in guinea pigs. However, no data are available regarding the expression and activity of the constitutive NO synthases (cNOS; NOS1 and NOS3, nNOS and eNOS, respectively) in this model. Therefore, we evaluated cNOS activity (conversion of L-[3H]arginine to L-[3H]citrulline in the presence of Ca2+ and calmodulin), nitrate and nitrite (NOx) concentration (modified Griess method), and NOS1 and NOS3 protein expression (Western blot) in lung homogenates and in the tracheal smooth muscle from OVA-immunized and multiple aerosol-challenged guinea pigs (six challenges, once daily). The expression and activity of the inducible NOS isoform (NOS2), the levels of exhaled NO, and the in vivo airway reactivity were also determined. Constitutive NOS activity and NO(x) concentration were significantly lower 6 h after the last OVA challenge as compared with saline exposure, being similar at 24 h. Expression of NOS1 paralleled cNOS activity, which was reduced 6, but not 24 h after OVA challenge. The decrease in NOS1 expression was accompanied by a significant decrease in the amounts of exhaled NO and by a maximal airway hyperresponsiveness to histamine. The levels of NOS3 were not modified at the two time points evaluated, and no NOS2 expression and activity were found at any time point. Similar modifications were observed in the tracheal smooth muscle. We conclude that OVA stimulation in immunized guinea pigs induced a transient reduction in NOS1 protein expression and activity in the respiratory system, which probably participates in airway hyperresponsiveness. PMID- 11435254 TI - Clara cell secretory protein oxidation and expression in premature infants who develop bronchopulmonary dysplasia. AB - Bronchopulmonary dysplasia (BPD) is a chronic lung disease that affects prematurely born infants and appears to evolve in part from early inflammatory responses in the lung. The inflammatory responses have been associated with protein and lipid oxidation in tracheal aspirate fluids. The present study was designed to test the hypothesis that in the first week of life specific oxidations and/ or altered expressions of proteins would be observed in tracheal aspirate fluids in infants who would subsequently develop BPD. We obtained tracheal aspirate fluids on Days of life 1, 3, and 6 from infants born at < or = 29 wk gestation, incubated the fluids with 2,4-dinitrophenylhyrazine (DNPH), separated the proteins electrophoretically, and assessed DNPH reactivity by immunonblots. DNPH reactivity of a protein that was identified as Clara cell secretory protein (CCSP) was observed more consistently in tracheal aspirate fluids from infants who later developed BPD than from infants who did not develop BPD. Tracheal aspirate fluid levels of immunoreactive CCSP were also lower on Day of life 1 in infants who developed BPD than in those who did not develop BPD. Increased CCSP oxidation and decreased immunoreactive CCSP expression in infants who subsequently developed BPD suggest that Clara cell function and CCSP expression may be critical for normal bronchoalveolar fluid homeostasis and that maintaining CCSP expression and function may be useful goals for targeted therapies for inhibition of the development of BPD. PMID- 11435255 TI - New serum markers for the detection of severe acute pancreatitis in humans. PMID- 11435256 TI - Role of the heart in the loss of aeration characterizing lower lobes in acute respiratory distress syndrome. PMID- 11435257 TI - EG-1 positive eosinophils in asthma. PMID- 11435258 TI - Short-course empiric antibiotic therapy for patients with pulmonary infiltrates in the intensive care unit. PMID- 11435259 TI - Case definition and culture: Are people all the same? PMID- 11435260 TI - Serotonin-based pharmacotherapy for acute neuroleptic-induced akathisia: a new approach to an old problem. PMID- 11435261 TI - What constitutes success? The relative priority of service users' and clinicians' views of mental health services. PMID- 11435262 TI - Somatoform disorders: severe psychiatric illnesses neglected by psychiatrists. AB - BACKGROUND: Somatoform disorders have few peers in terms of personal morbidity and cost to the health service, yet many psychiatrists train without any experience of them. AIMS: To review the prevalence, disability and economic burden of somatoform disorders, and to explore the reasons why they are neglected by psychiatrists. METHOD: A selective review of the key literature. RESULTS: Psychiatrists' current preoccupation with so-called 'serious mental illness' gives somatoform disorders low priority. Some health planners have erroneously equated severity with diagnosis rather than level of need and disability. As a consequence the development of psychiatric services has been neglected. CONCLUSIONS: Greater recognition of the importance of somatoform disorders will only occur if high quality research and teaching receive priority, and if the Royal Colleges continue to press for increasing public awareness of their importance. Services should be driven by clinical need rather than diagnosis. PMID- 11435263 TI - Venlafaxine extended release (ER) in the treatment of generalised anxiety disorder: twenty-four-week placebo-controlled dose-ranging study. AB - BACKGROUND: Generalised anxiety disorder (GAD) has received less study than other anxiety disorders, particularly its long-term treatment. AIMS: To assess the efficacy and safety of venlafaxine extended release (ER) in patients with GAD. METHOD: A total of 541 out-patients, 18-86 years old, were recruited to this 24 week, placebo-controlled, double-blind study of three fixed doses (37.5, 75 and 150 mg/day) of venlafaxine ER. RESULTS: All doses of venlafaxine ER showed efficacy superior to placebo, apparent from week 2, that was sustained throughout the 24-week study for the two higher doses. The discontinuation rate did not differ significantly among the treatment groups. CONCLUSIONS: Venlafaxine ER is an effective and safe treatment for GAD for up to 6 months. PMID- 11435264 TI - Randomised controlled general practice trial of sertraline, exposure therapy and combined treatment in generalised social phobia. AB - BACKGROUND: No controlled trial of treatment of generalised social phobia has been conducted in general practice. AIMS: To examine the efficacy of sertraline or exposure therapy, administered alone or in combination in this setting. METHOD: Study was of a randomised, double-blind design. Patients (n = 387) received sertraline 50-150 mg or placebo for 24 weeks. Patients were additionally randomised to exposure therapy or general medical care. RESULTS: Sertraline treated patients were significantly more improved than non-sertraline-treated patients (chi(2)=12.53, P<0.001; odds ratio=0.534; 95% Cl 0.347-0.835). No significant difference was observed between exposure- and non-exposure-treated patients (chi(2)=2.18, P=0.140; odds ratio=0.732; 95% Cl 0.475-1.134). In the pairwise comparisons, combined sertraline and exposure (chi(2)=12.32; P<0.001) and sertraline (chi(2)=10.13; P=0.002) were significantly superior to placebo. CONCLUSIONS: Sertraline is an effective treatment for generalised social phobia. Combined treatment with sertraline and exposure therapy, conducted by the general practitioner, may enhance the treatment efficacy in primary care. PMID- 11435265 TI - Seasonal affective disorder: prevalence, detection and current treatment in North Wales. AB - BACKGROUND: There is a paucity of information concerning the prevalence and detection of seasonal affective disorder (SAD) in UK populations. AIMS: To determine the prevalence, detection and current treatment of SAD within a general population sample. METHOD: The study was conducted in conjunction with the Outcomes of Depression International Network (ODIN) project, a large European study of depression. At the North Wales arm of the project, 1999 adults were randomly selected from a health authority database and screened by post for SAD with the Seasonal Patterns Assessment Questionnaire (SPAQ). Those scoring above cut-off were offered diagnostic interview, after which diagnosis of SAD according to DSM-IV criteria could be made. RESULTS: The prevalence rate of SAD was calculated to be 2.4% (95% CII.4-1.3). The majority of identified cases had not previously received a diagnosis of SAD from their general practitioner, although over half had been diagnosed with other forms of depression and had been prescribed antidepressant medication. CONCLUSIONS: Although SAD was found to be common in this general population sample it appeared to be largely underdiagnosed and/or misdiagnosed. PMID- 11435266 TI - Cigarette smoking and psychotic symptoms in bipolar affective disorder. AB - BACKGROUND: An association exists between smoking and schizophrenia, independent of other factors and related to psychotic symptomatology. AIMS: To determine whether smoking is associated with psychosis in bipolar affective disorder. METHOD: Smoking data were collected from 92 unrelated patients with bipolar affective disorder. An ordinal logistic regression analysis tested the relationship between smoking severity and psychotic symptomatology, allowing for potential confounders. RESULTS: A significant relationship was detected between smoking/heavy smoking and history of psychosis (68.7%, n=44). Smoking was less prevalent in patients who were less symptomatic (56.5%, n=13) than in patients with a more severe psychosis (75.7, n=31). Prevalence and severity of smoking predicted severity of psychotic symptoms (P=0.001), a relationship independent of other variables (P=0.0272). CONCLUSION: A link between smoking and psychosis exists in bipolar affective disorder and may be independent of categorical diagnosis. PMID- 11435267 TI - Twin study of symptom dimensions in psychoses. AB - BACKGROUND: Symptomatology in psychoses can be summarised as quantitative symptom dimensions, but their genetic basis is unknown. AIMS: To investigate whether genes make an important contribution to symptom dimensions. METHOD: A total of 224 probandwise twin pairs (106 monozygotic, 118 same-gender dizygotic) where probands had psychosis were ascertained from the Maudsley Twin Register in London. Factor analysis was performed on lifetime symptoms rated on the Operational Checklist for Psychotic Disorders (OPCRIT). Correlations of dimension scores within monozygotic and dizygotic pairs concordant for Research Diagnostic Criteria psychoses were performed. Relationships between dimension scores and genetic loading for psychoses were assessed using logistic regression. RESULTS: Patterns of familial aggregation consistent with a genetic effect were found for the disorganised dimension and for some measures of the negative, manic and general psychotic dimensions. Disorganised dimension scores were related significantly to genetic loading for psychoses. CONCLUSIONS: The disorganised dimension, and possibly other symptom dimensions, may be useful phenotypes for molecular genetic studies of psychoses. PMID- 11435268 TI - Family history, place and season of birth as risk factors for schizophrenia in Denmark: a replication and reanalysis. AB - BACKGROUND: Although a family history of schizophrenia is the strongest individual risk factor for schizophrenia, environmental factors related to urbanicity may contribute to a substantial proportion of the population occurrence of the disease. AIMS: This study replicates previous findings in four mutually exclusive Danish study populations, including out-patient information, ICD-10 diagnoses of schizophrenia, and a broader adjustment for mental illness in family members. METHOD: We established a population-based cohort of 2.66 million Danish people using data from the Civil Registration System linked with the Psychiatric Case Register. RESULTS: Overall, 10 264 persons developed schizophrenia during the 50.7 million person-years of follow-up. The risk of schizophrenia was increased by urbanicity of place of birth and by family history of schizophrenia or other mental disorders. CONCLUSIONS: Urban-rural differences of schizophrenia risk were replicated and could not be associated with the potential sources of bias we assessed. Environmental factors underlying the effect of place of birth are major determinants of schizophrenia occurrence at the population level, although the effect of family history is the strongest at the individual level. PMID- 11435269 TI - Social functioning and the course of early-onset schizophrenia: five-year follow up of a psychosocial intervention. AB - BACKGROUND: Schizophrenia implies severe social impairments. Since the treatment of patients with schizophrenia shifted from long-term hospital admissions to community services, research on social functioning has become increasingly important. AIMS: Follow-up assessment of social functioning in young patients with schizophrenia during a 5-year period after intervention. METHOD: During intervention, families were randomised into two conditions: standard intervention and standard plus family intervention. RESULTS: Although no differential treatment effect with regard to the course of the illness was found, patients from the standard plus family intervention condition stayed for fewer months in institutions for psychiatric patients than patients from the standard intervention condition. CONCLUSIONS: Family intervention has helped parents to support their children, thereby diminishing institutional care. PMID- 11435270 TI - Effects of long-term treatment with antipsychotics on serum leptin levels. AB - BACKGROUND: Abnormal regulation of the adipocyte-derived hormone leptin could play a role in body weight gain induced by antipsychotics. AIMS: To study the effects of long-term antipsychotic treatment on leptin levels in patients with schizophrenia. METHOD: Serum leptin levels were determined in 59 out-patients with chronic schizophrenia and in the same number of healthy subjects controlled by gender, age and body mass index. RESULTS: Leptin levels did not differ between patients and controls. Leptin levels in patients with schizophrenia correlated with weight gain, even after controlling for current weight, but did not show any association with clinical variables. Antipsychotic class tended to exert different effects over leptin levels (among atypicals, olanzapine induced a greater increase). CONCLUSIONS: Elevation of leptin levels induced by chronic antipsychotic treatment can be attributed to weight gain, but other mechanisms could be involved. PMID- 11435271 TI - Antipsychotic medication and venous thrombosis. AB - BACKGROUND: In an autopsy series, 10 out of 27 deaths in which 'idiopathic' pulmonary emboli were discerned as the sole cause of death had occurred in psychiatric patients. AIMS: To investigate whether antipsychotic medication is a risk factor for venous thrombosis. METHOD: A description of the 10 psychiatric patients was obtained from the pulmonary emboli autopsy reports. We carried out a brief historic overview of the literature. We re-analysed data from the Leiden Thrombophilia Study (LETS), a case-control study on patients with venous thrombosis. RESULTS: In the autopsy reports, five out of 10 psychiatric patients with fatal pulmonary embolism had confirmed use of antipsychotic drugs. After the application of chlorpromazine and its analogues a higher incidence of venous thrombosis in psychiatric patients was described in the German literature between 1953 and 1977. In the re-analysis of the LETS case-control study, four patients used antipsychotic drugs versus none in the control group. Recent epidemiological studies of good methodological quality have confirmed these findings. CONCLUSIONS: Venous thrombosis appears to be associated with the use of antipsychotic drugs in psychiatric patients. PMID- 11435272 TI - Carers' and Users' Expectations of Services--User version (CUES-U): a new instrument to measure the experience of users of mental health services. AB - BACKGROUND: No existing instrument measures all or even most of the issues considered important by users of mental health services. AIMS: To develop and test a self-assessment instrument to enable users of mental health services to rate their experience across the range of domains that they consider to be important. METHOD: Relevant domains were identified and a new instrument was drafted and field tested to examine its psychometric properties. RESULTS: The 17 item, self-rated Carers' and Users' Expectations of Services--User version (CUES U) appears acceptable to most service users. Its items have reasonable test retest reliability and a 'total CUES-U score' correlates significantly with a total score of the Health of the Nations Outcome Scales (Spearman's rho=0.42; P < 0.01). CONCLUSIONS: The development and testing of CUES-U suggest that it might be feasible to apply a self-rated measure of the expectations and experience of users of mental health services. PMID- 11435273 TI - Advising relatives of risk of Alzheimer's disease. PMID- 11435274 TI - Treatment for Alzheimer's disease in people with learning disabilities: NICE guidance. PMID- 11435275 TI - Community care for mental disorders in developing countries: a perspective. PMID- 11435276 TI - Mental health literacy in developing countries. PMID- 11435277 TI - Handedness and schizophrenia: genetic and environmental factors. PMID- 11435278 TI - Stress management and schizophrenia. PMID- 11435279 TI - Psychological therapies in anorexia nervosa. PMID- 11435280 TI - Psychological therapies in anorexia nervosa. PMID- 11435281 TI - Psychological therapies in anorexia nervosa. PMID- 11435282 TI - Does size matter? PMID- 11435283 TI - Quality of evidence in meta-analysis. PMID- 11435284 TI - Placebo response in depression. PMID- 11435286 TI - Personality disorder. PMID- 11435287 TI - Psychiatry in pictures. PMID- 11435291 TI - Platelets from patients with the Quebec platelet disorder contain and secrete abnormal amounts of urokinase-type plasminogen activator. AB - The Quebec platelet disorder (QPD) is an autosomal dominant platelet disorder associated with delayed bleeding and alpha-granule protein degradation. The degradation of alpha-granule, but not plasma, fibrinogen in patients with the QPD led to the investigation of their platelets for a protease defect. Unlike normal platelets, QPD platelets contained large amounts of fibrinolytic serine proteases that had properties of plasminogen activators. Western blot analysis, zymography, and immunodepletion experiments indicated this was because QPD platelets contained large amounts of urokinase-type plasminogen activator (u-PA) within a secretory compartment. u-PA antigen was not increased in all QPD plasmas, whereas it was increased more than 100-fold in QPD platelets (P <.00009), which contained increased u-PA messenger RNA. Although QPD platelets contained 2-fold more plasminogen activator inhibitor 1 (PAI-1) (P <.0008) and 100-fold greater u-PA PAI-1 complexes (P <.0002) than normal platelets, they contained excess u-PA activity, predominantly in the form of two chain (tcu-PA), which required additional PAI-1 for full inhibition. There was associated proteolysis of plasminogen in QPD platelets, to forms that comigrated with plasmin. When similar amounts of tcu-PA were incubated with normal platelet secretory proteins, many alpha-granule proteins were proteolyzed to forms that resembled degraded QPD platelet proteins. These data implicate u-PA in the pathogenesis of alpha-granule protein degradation in the QPD. Although patients with the QPD have normal to increased u-PA levels in their plasma, without evidence of systemic fibrinogenolysis, their increased platelet u-PA could contribute to bleeding by accelerating fibrinolysis within the hemostatic plug. QPD is the only inherited bleeding disorder in humans known to be associated with increased u-PA. PMID- 11435292 TI - Sudden death among patients with acute promyelocytic leukemia treated with arsenic trioxide. AB - Arsenic trioxide has been shown to be effective in treating acute promyelocytic leukemia (APL), with minimal overall toxicity reported to date. A phase I/II study was initiated in June 1998 using arsenic trioxide for relapsed APL to determine the maximum tolerated or minimal effective dose and to determine the efficacy of treatment at that dose. Ten patients received 1 to 4 monthly cycles of treatment with 0.1 mg/kg per day intravenous arsenic trioxide. Six of 7 patients evaluable for response achieved cytogenetic or molecular complete remission. However, 3 patients died suddenly during the first cycle of treatment. Autopsies obtained on 2 of these failed to identify a cause of sudden death, despite evidence of pulmonary hemorrhage in one. A third patient, for whom an autopsy was not performed, became asystolic and died while on continuous cardiac telemetry. These observations suggest that arsenic trioxide may be significantly or even fatally toxic at doses currently used and that caution is warranted in its use. PMID- 11435293 TI - Survival of transfused donor white blood cells in HIV-infected recipients. AB - The appearance and expansion of donor white blood cells in a recipient after transfusion has many potential biologic ramifications. Although patients with HIV infection are ostensibly at high risk for microchimerism, transfusion-associated graft-versus-host disease (TA-GVHD) is rare. The purpose of this study was to search for sustained microchimerism in such patients. Blood samples were collected from 93 HIV-infected women (a subset from the Viral Activation Transfusion Study, an NHLBI multicenter randomized trial comparing leukoreduced versus unmodified red blood cell [RBC] transfusions) before and after transfusions from male donors. Donor lymphocytes were detected in posttransfusion specimens using a quantitative Y-chromosome-specific polymerase chain reaction (PCR) assay, and donor-specific human leukocyte antigen (HLA) alleles were identified with allele-specific PCR primers and probes. Five of 47 subjects randomized to receive nonleukoreduced RBCs had detectable male lymphocytes 1 to 2 weeks after transfusion, but no subject had detectable male cells more than 4 weeks after a transfusion. In 4 subjects studied, donor-specific HLA haplotypes were detected in posttransfusion specimens, consistent with one or more donors' cells. None of 46 subjects randomized to receive leukoreduced RBCs had detectable male lymphocytes in the month after transfusion. Development of sustained microchimerism after transfusion in HIV-infected patients is rare; HIV-infected patients do not appear to be at risk for TA-GVHD. PMID- 11435294 TI - Clinical and virologic characteristics of chronic active Epstein-Barr virus infection. AB - Thirty patients with chronic active Epstein-Barr virus (CAEBV) infection were analyzed. The study group included 18 male and 12 female patients, ranging in age from 5 to 31 years with a mean age of 14.2 years. Not all patients had high titers of EBV-specific antibodies, but all patients had high viral loads in their peripheral blood (more than 10(2.5) copies/microg DNA). Fifty percent of the patients displayed chromosomal aberrations, and 79% had monoclonality of EBV. Patients were divided into 2 clinically distinct groups, based on whether the predominantly infected cells in their peripheral blood were T cells or natural killer (NK) cells. Over a 68-month period of observation, 10 patients died from hepatic failure, malignant lymphoma, or other causes. Patients with T-cell CAEBV had a shorter survival time than those with NK-cell type of disease. PMID- 11435295 TI - Development of herpes simplex virus-1 amplicon-based immunotherapy for chronic lymphocytic leukemia. AB - Herpes simplex virus (HSV)-based vectors have favorable biologic features for gene therapy of leukemia and lymphoma. These include high transduction efficiency, ability to infect postmitotic cells, and large packaging capacity. The usefulness of HSV amplicon vectors for the transduction of primary human B cell chronic lymphocytic leukemia (CLL) was explored. Vectors were constructed encoding beta-galactosidase (LacZ), CD80 (B7.1), or CD154 (CD40L) and were packaged using either a standard helper virus (HSVlac, HSVB7.1, and HSVCD40L) or a helper virus-free method (hf-HSVlac, hf-HSVB7.1, and hf-HSVCD40L). Both helper containing and helper-free vector stocks were studied for their ability to transduce CLL cells, up-regulate costimulatory molecules, stimulate allogeneic T cell proliferation in a mixed lymphocyte tumor reaction, and generate autologous cytotoxic T lymphocytes (CTLs). Although helper-containing and helper-free amplicon stocks were equivalent in their ability to transduce CLL cells, a vigorous T-cell proliferative response was obtained using cells transduced with hf-HSVB7.1 but not with HSVB7.1. CLL cells transduced with either HSVCD40L or hf HSVCD40L were compared for their ability to up-regulate resident B7.1 and to function as T-cell stimulators. Significantly enhanced B7.1 expression in response to CD40L was observed using hf-HSVCD40L but not with HSVCD40L. CLL cells transduced with hf-HSVCD40L were also more effective at stimulating T-cell proliferation than those transduced with HSVCD40L stocks and were successful in stimulating autologous CTL activity. It is concluded that HSV amplicons are efficient vectors for gene therapy of hematologic malignancies and that helper virus-free HSV amplicon preparations are better suited for immunotherapy. PMID- 11435296 TI - Regulation of hypoxia-inducible factor is preserved in the absence of a functioning mitochondrial respiratory chain. AB - Hypoxia-inducible factor (HIF) mediates a large number of transcriptional responses to hypoxia and has an important role in processes that include angiogenesis and erythropoiesis. The HIF DNA binding complex consists of 2 basic helix-loop-helix PAS proteins designated alpha and beta subunits. Regulation occurs principally through the alpha subunits, which are stabilized and activated in hypoxia. Although substantial evidence implicates reactive oxygen species (ROS) in the regulatory process, the precise mechanisms remain unclear. Mitochondria are an important source of ROS, and in one model it has been proposed that hypoxia increases the generation of ROS at complex III in the mitochondrion and that this signal acts through a transduction pathway to stabilize HIF-1alpha and to activate HIF. To test this model the induction of the HIF-1alpha subunit and the HIF target gene, glucose-transporter-1, was examined in a variety of mutant cells that lacked mitochondrial DNA (rho0) or had other genetic defects in mitochondrial respiration. HIF induction by hypoxia was essentially normal in all cells tested. Hydrogen peroxide production was measured by the luminol/peroxidase method and found to be reduced in rho0 versus wild-type cells and reduced by hypoxia in both rho0 and wild-type cells. Furthermore, concentrations of rotenone that maximally inhibited respiration did not affect HIF activation by hypoxia. These data do not support the model outlined above and indicate that a functional respiratory chain is not necessary for the regulation of HIF by oxygen. PMID- 11435297 TI - Correlation between nicotine-induced inhibition of hematopoiesis and decreased CD44 expression on bone marrow stromal cells. AB - This study demonstrates that in vivo exposure to cigarette smoke (CS) and in vitro treatment of long-term bone marrow cultures (LTBMCs) with nicotine, a major constituent of CS, result in inhibition of hematopoiesis. Nicotine treatment significantly delayed the onset of hematopoietic foci and reduced their size. Furthermore, the number of long-term culture-initiating cells (LTC-ICs) within an adherent layer of LTBMCs was significantly reduced in cultures treated with nicotine. Although the production of nonadherent mature cells and their progenitors in nicotine-treated LTBMCs was inhibited, this treatment failed to influence the proliferation of committed hematopoietic progenitors when added into methylcellulose cultures. Bone marrow stromal cells are an integral component of the hematopoietic microenvironment and play a critical role in the regulation of hematopoietic stem cell proliferation and self-renewal. Exposure to nicotine decreased CD44 surface expression on primary bone marrow-derived fibroblastlike stromal cells and MS-5 stromal cell line, but not on hematopoietic cells. In addition, mainstream CS altered the trafficking of hematopoietic stem/progenitor cells (HSPC) in vivo. Exposure of mice to CS resulted in the inhibition of HSPC homing into bone marrow. Nicotine and cotinine treatment resulted in reduction of CD44 surface expression on lung microvascular endothelial cell line (LEISVO) and bone marrow-derived (STR-12) endothelial cell line. Nicotine treatment increased E-selectin expression on LEISVO cells, but not on STR-12 cells. These findings demonstrate that nicotine can modulate hematopoiesis by affecting the functions of the hematopoiesis-supportive stromal microenvironment, resulting in the inhibition of bone marrow seeding by LTC-ICs and interfering with stem cell homing by targeting microvascular endothelial cells. PMID- 11435298 TI - Receptor binding cancer antigen expressed on SiSo cells, a novel regulator of apoptosis of erythroid progenitor cells. AB - To better understand the control of apoptosis during erythropoiesis, this study investigated the role of a novel tumor-associated antigen, RCAS1 (receptor binding cancer antigen expressed on SiSo cells), with regard to the regulation of apoptosis of erythroid progenitor cells. Erythroid colony-forming cells (ECFCs) purified from human peripheral blood were used. Binding experiments of RCAS1 showed that ECFCs abundantly expressed receptors (RCAS1R) for RCAS1 and that the degree of binding of RCAS1 to the receptors diminished rapidly during erythroid maturation in vitro. When the soluble form of RCAS1 was added to the cultures, ECFCs underwent apoptosis, including collapse of the mitochondrial transmembrane potential, and activation of caspases 8 and 3. The addition of an anti-Fas blocking antibody or Fas-Fc failed to reduce the apoptosis induced by RCAS1, thereby indicating that effects of RCAS1 are independent of Fas activation. When binding of RCAS1 to normal bone marrow cells was analyzed, RCAS1R was evident on cells with an immature erythroid phenotype (transferrin receptor(+)/glycophorin A(-)) but not with a mature phenotype (transferrin receptor(-)/glycophorin A(+)). Histochemical staining revealed the expression of RCAS1 in the cytoplasm of bone marrow macrophages. These findings indicate that RCAS1, which is mainly produced by macrophages in hematopoietic tissue, may have a crucial role in controlling erythropoiesis by modulating apoptosis of erythroid progenitor cells via a Fas independent mechanism. PMID- 11435299 TI - Hematopoietic responses to stress conditions in young dogs compared with elderly dogs. AB - Clinical observations show that older patients do not tolerate high-dose chemoradiotherapy as well as younger patients. It is unclear whether this is due to age-related differences in their responses to hematopoietic injury or to differential toxicities to other organs. In the present study, 6 young (0.5 years) and 6 elderly (8 years) dogs were challenged with 7 repeated nonlethal doses of 50 or 100 cGy total body irradiation (TBI) each (total 550 cGy), and 21 days of recombinant canine granulocyte-colony stimulating factor (rcG-CSF) after the last TBI dose. Recoveries of absolute neutrophil, platelet, and lymphocyte counts after each TBI dose, responses to rcG-CSF treatment, and telomere lengths in neutrophils were compared before and after the study. No differences were found in recoveries of neutrophils, platelets, or in responses to rcG-CSF among young and old dogs. In contrast, recoveries were suggestively worse in younger dogs. After rcG-CSF, platelet recoveries were poor in both groups compared with previous platelet recoveries (P <.01). Consequently, 2 old and 3 young dogs were euthanized because of persistent thrombocytopenia and bleeding. At the study's completion, marrow cellularities and peripheral blood counts of the remaining young and elderly dogs were equivalent. The telomere lengths in both groups were significantly reduced after the study versus beforehand (P =.03), but the median attritions of telomeres were not different. It was concluded that aging does not appear to affect hematopoietic cell recoveries after repeated low-dose TBI, suggesting that poor tolerance of radiochemotherapy regimens in older patients may be due to nonhematopoietic organ toxicities rather than age-related changes in hematopoietic stem cells reserves. PMID- 11435300 TI - Receptor specificity in the self-renewal and differentiation of primary multipotential hemopoietic cells. AB - To determine whether cytokine-induced signals generate unique responses in multipotential hemopoietic progenitor cells, the signaling domains of 3 different growth factor receptors (Mpl, granulocyte-colony-stimulating factor [G-CSF] receptor, and Flt-3) were inserted into mouse primary bone marrow cells. To circumvent the activation of endogenous receptors, each signaling domain was incorporated into an FK506 binding protein (FKBP) fusion to allow for its specific activation using synthetic FKBP ligands. Each signaling domain supported the growth of Ba/F3 cells; however, only Mpl supported the sustained growth of transduced marrow cells, with a dramatic expansion of multipotential progenitors and megakaryocytes. These findings demonstrate that the self-renewal and differentiation of multipotential progenitor cells can be influenced through distinct, receptor-initiated signaling pathways. PMID- 11435301 TI - Bone morphogenetic protein 4 induces efficient hematopoietic differentiation of rhesus monkey embryonic stem cells in vitro. AB - A cell culture system consisting of mouse S17 stromal cells supplemented with cytokines was developed for hematopoietic differentiation of rhesus monkey embryonic stem (ES) cells. The differentiated colonies that formed contained clusters of hematopoietic-like cells, as well as structures similar in appearance to embryonic blood islands. When this culture system was supplemented with bone morphogenetic protein 4 (BMP-4), the numbers of primary hematopoietic clusters increased by an average of 15 fold. The primary hematopoietic clusters containing clonogenic precursors (expandable hematopoietic clusters) increased by 18 fold. Immunofluorescence analysis showed that a substantial percentage of the hematopoietic-like cells were CD34(+), with morphologic features of undifferentiated blast cells. Enrichment of the CD34(+) cells was associated with enhanced stromal-dependent, cytokine-driven formation of cobblestone colonies on secondary plating. The hematopoietic identity of the precursors was further indicated by their expression of genes associated with hematopoietic differentiation, as well as morphologic assessments that showed erythroid and myeloid lineages among the progeny cells. In addition, reverse transcriptase polymerase chain reaction analysis of BMP-4-treated rhesus monkey ES cells demonstrated an up-regulation of early-expressed genes responsible for embryonic hematopoiesis and angiogenesis during the first 7 days of culture. These observations suggest that embryonic mesoderm regulatory protein may mimic physiologic signals that are required for the onset of embryonic hematopoiesis and stem cell formation in rhesus monkey ES cells. PMID- 11435302 TI - Lyn is required for normal stem cell factor-induced proliferation and chemotaxis of primary hematopoietic cells. AB - Stem cell factor (SCF) binds to c-Kit and is an important mediator of survival, growth, and function of hematopoietic progenitor cells and mast cells. Lyn and other Src family members are activated by SCF and associate with phosphorylated tyrosine residues in the c-Kit juxtamembrane region. However, studies using c-Kit mutants incapable of directly recruiting Src family members suggest this kinase family plays a minimal role in c-Kit stimulus-response coupling mechanisms. The objective of this study was to specifically target Lyn and subsequently address its role in SCF-mediated responses of primary hematopoietic progenitor cells and mast cells. To this end, a dominant-inhibitory Lyn mutant and Lyn-deficient mice were used. Transfection of normal murine mast cells with kinase-inactive Lyn impaired SCF-induced growth. Further, SCF-induced proliferation and chemotaxis of Lyn-deficient mast cells were less than for wild-type mast cells. SCF-induced growth of progenitor cells lacking Lyn was also reduced compared with that of wild-type progenitor cells. Impairment of SCF-mediated responses of Lyn-deficient mast cells and progenitor cells did not result from reductions in surface expression of c-Kit. These studies demonstrate that Lyn is required for normal SCF-mediated responses of primary progenitors and for a differentiated lineage. PMID- 11435303 TI - Fibrinogen Milano XII: a dysfunctional variant containing 2 amino acid substitutions, Aalpha R16C and gamma G165R. AB - Fibrinogen Milano XII was detected in an asymptomatic Italian woman, whose routine coagulation test results revealed a prolonged thrombin time. Fibrinogen levels in functional assays were considerably lower than levels in immunologic assays. Polymerization of purified fibrinogen was strongly impaired in the presence of calcium or ethylenediaminetetraacetic acid (EDTA). Two heterozygous structural defects were detected by DNA analysis: Aalpha R16C and gamma G165R. As seen previously with other heterozygous Aalpha R16C variants, thrombin-catalyzed release of fibrinopeptide A was 50% of normal. Additionally, the release of fibrinopeptide B was delayed. Immunoblotting analysis with antibodies to human serum albumin indicated that albumin is bound to Aalpha 16 C. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of plasmin digests of fibrinogen Milano XII in the presence of calcium or EDTA showed both normal and novel D1 and D3 fragments. Further digestion of abnormal D3 fragments by chymotrypsin resulted in degradation products of the same size as the fragments derived from normal fibrinogen. SDS-PAGE analysis under reducing conditions showed no difference between normal fibrinogen and fibrinogen Milano XII or between their plasmic fragments. Circular dichroism analysis revealed a shift in the mean residual ellipticity and a significant reduction of the alpha-helix content in the variant D3 fragment. It is concluded that the Aalpha-chain substitution is mainly responsible for the coagulation abnormalities, whereas the substitution in the gamma-chain induced a conformational change in the D3 fragment. PMID- 11435304 TI - Five novel mutations in the gene for human blood coagulation factor V associated with type I factor V deficiency. AB - Coagulation factor V (FV) plays an important role in maintaining the hemostatic balance in both the formation of thrombin in the procoagulant pathway as well as in the protein C anticoagulant pathway. FV deficiency is a rare bleeding disorder with variable phenotypic expression. Little is known about the molecular basis underlying this disease. This study identified 5 novel mutations associated with FV deficiency in 3 patients with severe FV deficiency but different clinical expression and 2 unaffected carriers. Four mutations led to a premature termination codon either by a nonsense mutation (single-letter amino acid codes): A1102T, K310Term. (FV Amersfoort) and C2491T, Q773Term. (FV Casablanca) or a frameshift: an 8-base pair deletion between nucleotides 1130 and 1139 (FV Seoul(1)) and a 1-base pair deletion between nucleotides 4291 and 4294 (FV Utrecht). One mutation was a novel missense mutation: T1927C, C585R (FV Nijkerk), resulting in the absence of mutant protein despite normal transcription to RNA. Most likely, an arginine at this position disrupts the hydrophobic interior of the FV A2 domain. The sixth detected mutation was a previously reported missense mutation: A5279G, Y1702C (FV Seoul(2)). In all cases, the presence of the mutation was associated with type I FV deficiency. Identifying the molecular basis of mutations underlying this rare coagulation disorder will help to obtain more insight into the mechanisms involved in the variable clinical phenotype of patients with FV deficiency. PMID- 11435305 TI - Increased thrombogenesis and embolus formation in mice lacking glycoprotein V. AB - The glycoprotein (GP) Ib-V-IX complex plays a critical role in initiating platelet adhesion to von Willebrand factor (vWF) at the site of vascular injury. The complex also forms a high-affinity binding site for thrombin. Using an intravital microscopy mouse model, it was previously established that vWF plays a critical role in mediating platelet adhesion and thrombus formation following mesenteric arteriolar injury induced by ferric chloride. Further characterization of this model showed that these thrombotic events were also thrombin dependent. Using this vWF- and thrombin-dependent model, this study shows that GP V gene deficiency significantly accelerates both platelet adhesion and thrombus formation in mice following arteriolar injury. The time required for vessel occlusion in GP V-deficient (GP V(-/-)) mice was significantly shorter than that in wild-type mice. Interestingly, large emboli were also produced in GP V(-/-) mice, but not in wild-type mice, causing frequent downstream occlusion. However, when the 2 genotypes were compared in the in vitro perfusion chamber where thrombin was inhibited by heparin, no significant differences were found in either initial single-platelet adhesion or thrombus volume. These results demonstrate that GP V(-/-) mice have accelerated thrombus growth in response to vascular injury and suggest that this is caused by enhanced thrombin-induced platelet activation rather than enhanced binding of GPIb-V-IX to vWF. Absence of GP V also compromises thrombus stability. PMID- 11435306 TI - The functional interactions between CD98, beta1-integrins, and CD147 in the induction of U937 homotypic aggregation. AB - CD98 is expressed on both hematopoietic and nonhematopoietic cells and has been implicated in a variety of different aspects of cell physiology and immunobiology. In this study, the functional interactions between CD98 and other adhesion molecules on the surface of the promonocyte line U937 are examined by means of a quantitative assay of cell aggregation. Several of the CD98 antibodies induced homotypic aggregation of these cells without affecting cellular viability or growth. Aggregation induced by CD98 antibodies could be distinguished from that induced by beta1-integrin (CD29) ligation by lack of sensitivity to EDTA and by increased sensitivity to deoxyglucose. Aggregation induced via CD98 and CD29 could also be distinguished by the pattern of protein tyrosine phosphorylation induced. Some CD29 antibodies partially inhibited CD98-induced aggregation, and these antibodies were neither agonistic for aggregation nor inhibitors of beta1 integrin binding to substrates. Conversely, some CD98 antibodies were potent inhibitors of CD29-induced aggregation. Antibodies to beta2 integrins also partially inhibited CD98-induced aggregation. Unexpectedly, 2 antibodies to CD147, an immunoglobulin superfamily member whose function has remained unclear, were also potent inhibitors of both the aggregation and the protein tyrosine phosphorylation induced via CD98 ligation. The results of this study support a central role for CD98 within a multimolecular unit that regulates cell aggregation. PMID- 11435307 TI - The HIV protease inhibitor Indinavir inhibits cell-cycle progression in vitro in lymphocytes of HIV-infected and uninfected individuals. AB - Indinavir (IDV) is a potent and selective human immunodeficiency virus type 1 (HIV-1) protease inhibitor (PI) widely used in antiretroviral therapy for suppression of HIV, but its effects on the immune system are relatively unknown. Recently, it has been reported that PIs inhibit lymphocyte apoptosis. In the present study we have investigated the effects of ex vivo addition of IDV on lymphocyte activation and apoptosis in cells from HIV-infected children (n = 18) and from healthy uninfected individuals (controls, n = 5) as well as in Jurkat and PM1 T-cell lines. Pretreatment of control peripheral blood mononuclear cell (PBMC) cultures with IDV resulted in a dose-dependent inhibition of lymphoproliferative responses to different activation stimuli. Additionally, this treatment led to cell-cycle arrest in G0/G1 phase in anti-CD3 monoclonal antibody stimulated PBMC cultures in controls and in 15 of 18 HIV-infected children. Spontaneous- or activation-induced apoptosis of PBMCs from HIV-infected or uninfected individuals or of Fas-induced apoptosis in Jurkat and PM1 T cell lines were not inhibited by IDV. Moreover, IDV did not inhibit activation of caspases 1, -3, -4, -5, -9, and -8 in lysates of Jurkat T cells undergoing Fas-induced apoptosis. The findings indicate that IDV interferes with cell-cycle progression in primary cells but does not directly affect apoptosis. It is concluded that IDV may prolong cell survival indirectly by inhibiting their entry into cell cycle. In individuals on PI therapy, PI-mediated effects could potentially modulate immunologic responses independently of antiviral activity against HIV. PMID- 11435308 TI - Major histocompatibility complex-mismatched allogeneic bone marrow transplantation using perforin and/or Fas ligand double-defective CD4(+) donor T cells: involvement of cytotoxic function by donor lymphocytes prior to graft versus-host disease pathogenesis. AB - Experimental allogeneic bone marrow transplantation (BMT) models using cytotoxic single-deficient (perforin/granzyme or Fas ligand [FasL]) and cytotoxic double deficient (cdd) CD4(+) donor T cells have previously demonstrated roles for both effector pathways in graft-versus-host disease (GVHD). In the present study, the role of CD4-mediated antihost cytotoxicity in a GVH response is further examined across a complete major histocompatibility complex class I/II mismatch. As predicted, a double cytotoxic deficiency resulted in a clear delay in GVH associated weight loss, clinical changes, and mortality. Interestingly, analysis of donor T-cell presence in 5.5-Gy recipients soon after BMT demonstrated that the double cytotoxic deficiency resulted in a marked decrease in donor CD4 numbers. Transplantation of singularly perforin- or FasL-deficient donor CD4(+) T cells demonstrated that the absence of FasL was responsible for the markedly diminished CD4 number in recipient lymph nodes and spleens soon after BMT. However, increasing recipient total body irradiation conditioning (11.0 Gy) abrogated the decrease in FasL-defective B6-cdd and B6-gld CD4 numbers. Thus, the decrease was not a result of inherent CD4 defects, but was probably attributable to host resistance. Consistent with these observations, transplantation into 11.0 Gy recipients resulted in identical GVH lethality by equal numbers of B6 wild type, B6-cdd, and B6-gld CD4(+) T-cell inoculum. In total, the findings indicate that aggressive host conditioning lessens the requirement for donor CD4(+) cytotoxic function in GVH responses soon after BMT. The present results thus support the notion of a role for cytotoxic effector function in donor CD4(+) T cells prior to GVH-induced tissue injury. PMID- 11435309 TI - Impaired progenitor cell function in HIV-negative infants of HIV-positive mothers results in decreased thymic output and low CD4 counts. AB - Hematologic and immunologic functions were examined in 19 HIV-negative infants of HIV-positive mothers and 19 control infants of HIV-negative mothers. Control infants were selected to match for gestational age, weight, and mode of delivery. Cord blood was obtained from all infants and used for flow cytometric determination of lymphocyte subsets, including the naive CD4 count. Furthermore, to determine thymic output, cord blood mononuclear cells were used for determination of T-cell receptor excision circles (TRECs). Evaluation of progenitor cell function was done by means of colony-forming cell assay and fetal thymic organ cultures (FTOCs). Lower naive CD4 counts (459.3 +/- 68.9 vs 1128.9 +/- 146.8 cells/microL, P <.001) and reduced thymic output in infants of HIV positive mothers were found (frequency of CD4(+) cells with TRECs was 3.6% +/- 0.7% compared with 14.3% +/- 2.2% in controls, P <.001). In combination with lower red blood cell counts in infants of HIV-positive mothers, this finding suggested impairment of progenitor cell function. Indeed, progenitors from infants of HIV-positive mothers had decreased cloning efficiency (15.7% +/- 2.6% vs 55.8% +/- 15.9%, P =.009) and seemed to generate fewer T cells in FTOCs. In conclusion, lower numbers of naive CD4(+) cells and reduced thymic output in HIV negative infants of HIV-positive mothers may be due to impaired progenitor cell function. PMID- 11435310 TI - Mitochondrial membrane sensitivity to depolarization in acute myeloblastic leukemia is associated with spontaneous in vitro apoptosis, wild-type TP53, and vicinal thiol/disulfide status. AB - Nonresponse to remission-induction chemotherapy, which remains a major problem in acute myeloblastic leukemia (AML), has been linked to cellular resistance to apoptosis. Because the apoptosis induced by chemotherapeutic drugs is mediated by loss of mitochondrial transmembrane potential (MTP), it was postulated that sensitivity to mitochondrial membrane depolarization might be heterogeneous in AML. Using the uncoupling agent carbonyl cyanide m-chlorophenylhydrazone (mClCCP), the mitochondrial membrane sensitivity to depolarization (mClCCP concentrations that inhibit 50% of the transmembrane potential [IC(50)]) in AML blasts was measured and demonstrated marked interclonal heterogeneity, with the existence of comparatively sensitive (median mClCCP IC(50), 4 microM) and resistant (median mClCCP IC(50), 10 microM) clones. Furthermore, the mClCCP IC(50) was inversely associated with spontaneous in vitro apoptosis (P =.001). It was high in cases with mutant TP53 and correlated with the total cellular level of the multidrug resistance-associated protein (P =.019) but not of bcl-2, bax, or bcl-x. It was also found that the dithiol oxidant diamide, in contrast to the monovalent thiol oxidant diethyl maleate, increased the sensitivity of mitochondrial membranes to mClCCP. To confirm that TP53 directly affects MTP in leukemic cells and to establish the role of vicinal thiol oxidation in the TP53 dependent pathway, CEM 4G5 leukemia cells with forced, temperature-dependent expression of TP53 were studied. Monobromobimane, which inhibits mitochondrial membrane depolarization by preventing dithiol cross-linking, inhibited depolarization and apoptosis in 4G5 cells. It was concluded that in leukemia, TP53 and vicinal thiol/disulfide status are determinants of mitochondrial membrane sensitivity to depolarization, which is in turn associated with spontaneous apoptosis. PMID- 11435311 TI - Apaf-1 protein deficiency confers resistance to cytochrome c-dependent apoptosis in human leukemic cells. AB - The human leukemia cell lines K562, CEM, CEM/VLB(100), human leukemic blasts, and the bladder cancer J82 cell line have different sensitivities to UV light-induced apoptosis. It is reported that resistance to UV light-induced apoptosis occurs at a point in the apoptotic pathway upstream of caspase-3 but downstream of mitochondrial cytochrome c release. It is demonstrated that the block is due to deficiency of Apaf-1, a critical member of the apoptosome. Sensitivity to apoptosis was independent of caspase-9b or XIAP (inhibitors of apoptosis proteins) expression or levels of procaspase-9. Transfection of Apaf-1 conferred sensitivity to apoptosis in resistant cells. Apaf-1 deficiency may constitute a significant mode of resistance to apoptosis in human leukemia. PMID- 11435312 TI - Identification of myelodysplastic syndrome-specific genes by DNA microarray analysis with purified hematopoietic stem cell fraction. AB - Myelodysplastic syndrome (MDS) is a slowly progressing hematologic malignancy associated with a poor outcome. Despite the relatively high incidence of MDS in the elderly, differentiation of MDS from de novo acute myeloid leukemia (AML) still remains problematic. Identification of genes expressed in an MDS-specific manner would allow the molecular diagnosis of MDS. Toward this goal, AC133 surface marker-positive hematopoietic stem cell (HSC)-like fractions have been collected from a variety of leukemias in a large-scale and long-term genomics project, referred to as "Blast Bank," and transcriptome of these purified blasts from the patients with MDS were then compared with those from AML through the use of oligonucleotide microarrays. A number of genes were shown to be expressed in a disease-specific manner either to MDS or AML. Among the former found was the gene encoding the protein Delta-like (Dlk) that is distantly related to the Delta Notch family of signaling proteins. Because overexpression of Dlk may play a role in the pathogenesis of MDS, the disease specificity of Dlk expression was tested by a quantitative "real-time" polymerase chain reaction analysis. Examination of the Blast Bank samples from 22 patients with MDS, 31 with AML, and 8 with chronic myeloid leukemia confirmed the highly selective expression of the Dlk gene in the individuals with MDS. Dlk could be the first candidate molecule to differentiate MDS from AML. The proposal is made that microarray analysis with the Blast Bank samples is an efficient approach to extract transcriptome data of clinical relevance for a wide range of hematologic disorders. PMID- 11435313 TI - Vascular endothelial growth factor triggers signaling cascades mediating multiple myeloma cell growth and migration. AB - Multiple myeloma (MM) remains incurable, with a median survival of 3 to 4 years. This study shows direct effects of vascular endothelial growth factor (VEGF) upon MM and plasma cell leukemia (PCL) cells. The results indicate that VEGF triggers tumor cell proliferation via a protein kinase C (PKC)-independent Raf-1-MEK extracellular signal-regulated protein kinase pathway, and migration via a PKC dependent pathway. These observations provide the framework for novel therapeutic strategies targeting VEGF signaling cascades in MM. PMID- 11435314 TI - Improved superoxide-generating ability by interferon gamma due to splicing pattern change of transcripts in neutrophils from patients with a splice site mutation in CYBB gene. AB - Chronic granulomatous disease (CGD) is an inherited disorder of host defense against microbial infections caused by defective activity of the phagocyte NADPH oxidase. Based on an increase of neutrophil superoxide-generating ability in response to interferon gamma (IFN-gamma) in a single patient with CGD, multicentered group studies demonstrated a beneficial effect of prophylactic IFN gamma. However, no apparent increase of the phagocyte superoxide generation was found in patients enrolled in these studies. The present report offers an additional kindred in whom an IFN-gamma-dependent increase in neutrophil superoxide production was observed in 3 affected patients. The defect in the CYBB gene for gp91-phox was identified as an otherwise silent mutation adjacent to the third intron of the CYBB gene that alters messenger RNA splicing. By molecular analysis, significant differences were found in the splicing pattern of CYBB gene transcripts in patient neutrophils between 1 and 25 days after administration of IFN-gamma. Furthermore, a complete transcript containing the missing exons could be detected in all specimens after the treatment. The changes in the splicing pattern of the transcripts and the prolonged effect on superoxide-generating ability of patient neutrophils indicate that IFN-gamma induced a partial correction of the abnormal splicing of CYBB gene transcripts in myeloid progenitor cells. PMID- 11435315 TI - Targeted deletion of the CD59 gene causes spontaneous intravascular hemolysis and hemoglobinuria. AB - The glycolipid-anchored glycoprotein CD59 inhibits assembly of the lytic membrane attack complex of complement by incorporation into the forming complex. Absence of CD59 and other glycolipid-anchored molecules on circulating cells in the human hemolytic disorder paroxysmal nocturnal hemoglobinuria is associated with intravascular hemolysis and thrombosis. To examine the role of CD59 in protecting host tissues in health and disease, CD59-deficient (CD59(-/-)) mice were produced by gene targeting in embryonic stem cells. Absence of CD59 was confirmed by staining cells and tissues with specific antibody. Despite the complete absence of CD59, mice were healthy and fertile. Erythrocytes in vitro displayed increased susceptibility to complement and were positive in an acidified serum lysis test. Despite this, CD59(-/-) mice were not anemic but had elevated reticulocyte counts, indicating accelerated erythrocyte turnover. Fresh plasma and urine from CD59(-/-) mice contained increased amounts of hemoglobin when compared with littermate controls, providing further evidence for spontaneous intravascular hemolysis. Intravascular hemolysis was increased following administration of cobra venom factor to trigger complement activation. CD59(-/-) mice will provide a tool for characterizing the importance of CD59 in protection of self tissues from membrane attack complex damage in health and during diseases in which complement is activated. PMID- 11435316 TI - A comparison of the in vivo kinetics of Plasmodium falciparum ring-infected erythrocyte surface antigen-positive and -negative erythrocytes. AB - Ring-infected erythrocyte surface antigen (RESA)-positive, Plasmodium falciparum negative red blood cells (RBCs) are cells from which the malaria parasite has been removed by the host without the destruction of the erythrocyte ("pitting"). The survival of RESA-RBCs in vivo was assessed in 14 severe and 6 uncomplicated falciparum malaria patients. The mean RESA-RBC life of 183 hours (95% confidence interval [CI], 136-246) was longer than the median parasite clearance time of 66 hours (range, 30-108 hours) but shorter than the mean red cell life of 1027 hours (95% CI, 840-1213) (P =.0004), with a median ratio of 0.2:1.0 (range, 0.1-0.7). The estimated median percentage of parasites pitted/body transit was 0.003% (range, 0.001%-0.05%). The rate of rise of the RESA-RBC count during the first 24 hours after antimalarial treatment was significantly faster (P =.036) and the subsequent RESA-RBC survival significantly shorter (P =.017) after treatment with an artemisinin derivative than after treatment with quinine. Parasitization of red cells leads to changes in the erythrocyte that shorten their survival even if the parasite is removed subsequently. PMID- 11435317 TI - Intercellular adhesion molecule-4 binds alpha(4)beta(1) and alpha(V)-family integrins through novel integrin-binding mechanisms. AB - The LW blood group glycoprotein, ICAM-4, is a member of the intercellular adhesion molecule (ICAM) family expressed in erythroid cells. To begin to address the function of this molecule, ligands for ICAM-4 on hemopoietic and nonhemopoietic cell lines were identified. Peptide inhibition studies suggest that adhesion of cell lines to an ICAM-4-Fc construct is mediated by an LDV inhibitable integrin on hemopoietic cells and an RGD-inhibitable integrin on nonhemopoietic cells. Antibody inhibition studies identified the hemopoietic integrin as alpha(4)beta(1.) Antibody inhibition studies on alpha(4)beta(1) negative, nonhemopoietic cell lines suggested that adhesion of these cells is mediated by alpha(V) integrins (notably alpha(V)beta(1) and alpha(V)beta(5)). The structure of ICAM-4 modeled on the crystal structure of ICAM-2 was used to identify surface-exposed amino acid residues for site-directed mutagenesis. Neither an unusual LETS nor an LDV motif in the first domain of ICAM-4 was critical for integrin binding. ICAM-4 is the first ICAM family member shown to be a ligand for integrins other than those of the beta(2) family, and the data suggest that ICAM-4 has a novel integrin-binding site(s). These findings suggest a role for ICAM-4 in normal erythropoiesis and may also be relevant to the adhesive interactions of sickle cells. PMID- 11435318 TI - Requirements for the promotion of allogeneic engraftment by anti-CD154 (anti CD40L) monoclonal antibody under nonmyeloablative conditions. AB - The promotion of alloengraftment in the absence of global immune suppression and multiorgan toxicity is a major goal of transplantation. It is demonstrated that the infusion of a single modest bone marrow dosage in 200 cGy-irradiated recipients treated with anti-CD154 (anti-CD40L) monoclonal antibody (mAb) resulted in chimerism levels of 48%. Reducing irradiation to 100 or 50 cGy permitted 24% and 10% chimerism, respectively. In contrast, pan-T-cell depletion resulted in only transient engraftment in 200 cGy-irradiated recipients. Host CD4(+) cells were essential for alloengraftment as depletion of CD4(+) cells abrogated engraftment in anti-CD154-treated recipients. Strikingly, the depletion of CD8(+) cells did not further enhance engraftment in anti-CD154 mAb-treated recipients in a model in which rejection is mediated by both CD4(+) and CD8(+) T cells. However, anti-CD154 mAb did facilitate engraftment in a model in which only CD8(+) T cells mediate rejection. Furthermore, CD154 deletional mice irradiated with 200 cGy irradiation were not tolerant of grafts, suggesting that engraftment promotion by anti-CD154 mAb may not simply be the result of CD154:CD40 blockade. Together, these data suggest that a CD4(+) regulatory T cell may be induced by anti-CD154 mAb. In contrast to anti-CD154 mAb, anti-B7 mAb did not promote donor engraftment. Additionally, the administration of either anti CD28 mAb or anti-CD152 (anti-CTLA-4) mAb or the use of CD28 deletional recipients abrogated engraftment in anti-CD154 mAb-treated mice, suggesting that balanced CD28/CD152:B7 interactions are required for the engraftment-promoting capacity of anti-CD154 mAb. These data have important ramifications for the design of clinical nonmyeloablative regimens based on anti-CD154 mAb administration. PMID- 11435319 TI - The human erythropoietin receptor gene rescues erythropoiesis and developmental defects in the erythropoietin receptor null mouse. AB - Erythropoietin and its receptor are required for definitive erythropoiesis and maturation of erythroid progenitor cells. Mice lacking the erythropoietin receptor exhibit severe anemia and die at about embryonic day 13.5. This phenotype can be rescued by the human erythropoietin receptor transgene. Animals expressing only the human erythropoietin receptor survived through adulthood with normal hematologic parameters and appeared to respond appropriately to induced anemic stress. In addition to restoration of erythropoiesis during development, the cardiac defect associated with embryos lacking the erythropoietin receptor was corrected and the increased apoptosis in fetal liver, heart, and brain in the erythropoietin receptor null phenotype was markedly reduced. These studies indicate that no species barrier exists between mouse and human erythropoietin receptor and that the human erythropoietin receptor transgene is able to provide specific expression in hematopoietic and other selected tissues to rescue erythropoiesis and other organ defects observed in the erythropoietin receptor null mouse. PMID- 11435320 TI - Molecular tracking of leukemogenesis in a triplet pregnancy. AB - The occurrence of childhood acute lymphoblastic leukemia (ALL) in 2 of 3 triplets provided a unique opportunity for the investigation of leukemogenesis and the natural history of ALL. The 2 leukemic triplets were monozygotic twins and shared an identical, acquired TEL-AML1 genomic fusion sequence indicative of a single cell origin in utero in one fetus followed by dissemination of clonal progeny to the comonozygotic twin by intraplacental transfer. In accord with this interpretation, clonotypic TEL-AML1 fusion sequences could be amplified from the archived neonatal blood spots of the leukemic twins. The blood spot of the third, healthy, dizygotic triplet was also fusion gene positive in a single segment, though at age 3 years, his blood was found negative by sensitive polymerase chain reaction (PCR) screening for the genomic sequence and by reverse transcription PCR. Leukemic cells in both twins had, in addition to TEL-AML1 fusion, a deletion of the normal, nonrearranged TEL allele. However, this genetic change was found by fluorescence in situ hybridization to be subclonal in both twins. Furthermore, mapping of the genomic boundaries of TEL deletions using microsatellite markers indicated that they were individually distinct in the twins and therefore must have arisen as independent and secondary events, probably after birth. These data support a multihit temporal model for the pathogenesis of the common form of childhood leukemia. PMID- 11435321 TI - Acquired pure red cell aplasia associated with lymphoproliferative disease of granular T lymphocytes. AB - Acquired pure red cell aplasia (PRCA) can be associated with lymphoproliferative disease of granular T lymphocytes (T-LDGL), also known as T-cell large granular lymphocyte leukemia. Fifteen adult patients with PRCA associated with T-LDGL comprise this study. Neutropenia and rheumatoid arthritis were uncommon. All patients responded to immunosuppressive therapy. The 2 most commonly used treatments were prednisone and cyclophosphamide +/- corticosteroids, producing overall response rates of 50% and 60%, respectively. Treatment with cyclophosphamide was associated with a more durable remission (median, 60 versus 7.5 months). After a median follow-up of 67 months, 2 patients died of treatment related complications, one from myelodysplasia and another from cyclosporine induced renal failure. The clinical course and treatment responses of PRCA associated with T-LDGL in this series were similar to the general group of PRCA. Because T-LDGL is frequently underdiagnosed, it is likely that a significant proportion of idiopathic or primary PRCA is in fact secondary to T-LDGL. PMID- 11435322 TI - Low frequency of BCL-2/J(H) translocation in peripheral blood lymphocytes of healthy Japanese individuals. AB - The incidence of follicular lymphoma differs significantly between white and Japanese individuals. Translocation between the BCL-2 and immunoglobulin heavy chain genes is detected in 85% to 90% of all follicular lymphomas in whites. Recently, BCL-2/J(H) translocation was detected in peripheral blood lymphocytes from more than 50% of healthy white individuals. To clarify the reason for the difference in incidence of follicular lymphoma between whites and Japanese, the frequency of BCL-2/J(H) translocation in peripheral blood lymphocytes of healthy Japanese individuals was compared with that of German individuals. The prevalence of BCL-2/J(H) translocation in Japanese adults appeared to be significantly lower than that in German adults. The present data suggest that the low frequency of BCL-2/J(H) translocation in the Japanese general population may be one of the major reasons for the difference in incidence of follicular lymphoma between whites and Japanese. PMID- 11435323 TI - Factors influencing B lymphopoiesis after allogeneic hematopoietic cell transplantation. AB - In 93 allograft recipients, the numbers of marrow B-cell precursors on days 80 and 365 correlated with the counts of circulating B cells, suggesting that the posttransplantation B-cell deficiency is at least in part due to insufficient B lymphopoiesis. Factors that could affect B lymphopoiesis were evaluated. The number of marrow B-cell precursors on days 30 and 80 was at least 4-fold lower in patients with grade 2 to 4 acute graft-versus-host disease (GVHD) compared with patients with grade 0 to 1 acute GVHD. The number of B-cell precursors on day 365 was 18-fold lower in patients with extensive chronic GVHD compared with patients with no or limited chronic GVHD. The number of B-cell precursors was not related to CD34 cell dose, type of transplant (marrow versus blood stem cells), donor age, or patient age. It was concluded that posttransplantation B-cell deficiency results in part from inhibition of B lymphopoiesis by GVHD and/or its treatment. PMID- 11435324 TI - Extended survival in advanced and refractory multiple myeloma after single-agent thalidomide: identification of prognostic factors in a phase 2 study of 169 patients. AB - This report of a phase 2 trial of thalidomide (THAL) (200 mg/d; 200 mg increment every 2 weeks to 800 mg) for 169 patients with advanced myeloma (MM) (abnormal cytogenetics (CG), 67%; prior autotransplant, 76%) extends earlier results in 84 patients. A 25% myeloma protein reduction was obtained in 37% of patients (50% reduction in 30% of patients; near-complete or complete remission in 14%) and was more frequent with low plasma cell labeling index (PCLI) (below 0.5%) and normal CG. Two-year event-free and overall survival rates were 20% +/- 6% and 48% +/- 6%, respectively, and these were superior with normal CG, PCLI of less than 0.5%, and beta(2)-microglobulin of 3 mg/L. Response rates were higher and survival was longer especially in high-risk patients given more than 42 g THAL in 3 months (median cumulative dose) (landmark analysis); this supports a THAL dose-response effect in advanced MM. PMID- 11435325 TI - Expression of p15(ink4b) gene during megakaryocytic differentiation of normal and myelodysplastic hematopoietic progenitors. AB - In myelodysplastic syndrome (MDS), the expression of the cyclin-dependent kinase inhibitor p15(ink4B) (p15) is frequently decreased because of the aberrant methylation of the gene promoter; p15 is normally up-regulated during megakaryocytic differentiation. It was hypothesized that p15 methylation and deregulation of gene expression contribute to defective megakaryocytopoiesis in patients with MDS. Here it is shown that the increasing autocrine production of TGF-beta1 stimulates megakaryocytic differentiation in normal CD34(+) cells and that p15 mediates, at least in part, this effect. This TGF-beta1-dependent pathway is altered in MDS CD34(+) progenitors because of p15 methylation. The demethylating agent 2-deoxyAZAcytidin can restore the normal demethylated state of the p15 gene and increase its expression. Nevertheless, MDS CD34(+) cells only poorly differentiate to the megakaryocytic lineage. These findings suggest that p15 methylation occurs in a neoplastic clone with a profound defect of cell proliferation, survival, and differentiation that cannot be overcome by using a demethylating drug. PMID- 11435326 TI - Carotid filters: new additions to the interventionist's toolbox. PMID- 11435327 TI - Inflammatory cardiomyopathy (myocarditis): which patients should be treated with anti-inflammatory therapy? PMID- 11435328 TI - Syncope evaluation at a crossroad: for which patients? PMID- 11435329 TI - Calcineurin inhibition in hypertrophy: back from the dead! PMID- 11435330 TI - Cerebral protection with filter devices during carotid artery stenting. AB - BACKGROUND: Distal embolization of debris during percutaneous carotid artery stenting may result in neurological deficit. Filter devices for cerebral protection potentially reduce the risk of embolization. METHODS AND RESULTS: Elective carotid stent implantation using 3 different types of distal filter protection devices was attempted in 88 consecutive lesions (84 patients) in the internal carotid artery that had >70% diameter stenosis (mean, 78.7+/-10.7%). Procedures were performed in 3 different centers. The mean age of the patients was 69+/-8 years, 75% were men, and 35.7% had neurological symptoms. In 86 lesions, a stent was successfully implanted (97.7%). In 83 of these 86 procedures (96.5%), it was possible to position a filter device. In 53% of filters, there was macroscopic evidence of debris. Collected material consisted of lipid-rich macrophages, fibrin material, and cholesterol clefts. Neurological complications during the procedure, in the hospital, and at 30 days of clinical follow-up occurred in only one patient (1.2%). This patient suffered a minor stroke that resolved within 1 week. Two major adverse cardiac events (2.3%) occurred during the 30 days of follow-up. CONCLUSIONS: Filter protection during carotid artery stenting seems feasible and safe. In the present series, the incidence of neurological complications was low. PMID- 11435331 TI - Synergistic effect of urotensin II with mildly oxidized LDL on DNA synthesis in vascular smooth muscle cells. AB - BACKGROUND: The urotensin II (UII) found in coronary atheroma is the most potent vasoconstrictor known to date. Mildly oxidized LDL (moxLDL) contributes to atherogenesis and plaque formation. We assessed the effect of UII and its interaction with moxLDL and the oxidative components of moxLDL on vascular smooth muscle cell (VSMC) proliferation. Methods and Results-Growth-arrested VSMCs were incubated in serum-free medium with different concentrations of LDL, moxLDL, oxLDL, hydrogen peroxide, lysophosphatidylcholine, or 4-hydroxy-2-nonenal, with or without UII. [(3)H]Thymidine incorporation into DNA was measured as an index of VSMC proliferation. UII stimulated [(3)H]thymidine incorporation in a dose dependent manner, with a maximal effect at a concentration of 50 nmol/L (161%). Low concentrations of UII potentiated the mitogenic effect of LDL (108% to 242%), oxLDL (129% to 302%), moxLDL (120% to 337%), hydrogen peroxide (177% to 226%), lysophosphatidylcholine (115% to 332%), and 4-hydroxy-2-nonenal (142% to 299%). The synergistic interaction between UII and moxLDL was partially inhibited by anti-Gq/11alpha antibody, the epidermal growth factor receptor tyrosine kinase inhibitor erbstatin A (10 micromol/L), and the intracellular free radical scavenger N-acetylcysteine (400 micromol/L) and was completely inhibited by the c Src tyrosine kinase inhibitor radicicol (10 micromol/L), the protein kinase C (PKC) inhibitor Ro31-8220 (0.1 micromol/L), and the mitogen-activated protein kinase (MAPK) kinase inhibitor PD098059 (10 micromol/L). CONCLUSIONS: Our results suggest that UII acts synergistically with moxLDL in inducing VSMC proliferation via the c-Src/PKC/MAPK pathway, which may explain the relatively rapid progression of atherosclerosis in patients with hypertension and hypercholesterolemia. PMID- 11435332 TI - Trends in acute coronary heart disease mortality, morbidity, and medical care from 1985 through 1997: the Minnesota heart survey. AB - BACKGROUND: Coronary heart disease (CHD) mortality continued to decline from 1985 to 1997. METHODS AND RESULTS: We tabulated CHD deaths (ICD-9 codes 410 through 414) in the Minneapolis/St Paul, Minnesota, area. For 1985, 1990, and 1995, trained nurses abstracted the hospital records of patients 30 to 74 years old with a discharge diagnosis of acute CHD (ICD-9 codes 410 or 411). Acute myocardial infarction (AMI) events were validated and followed for 3-year all cause mortality. Between 1985 and 1997, age-adjusted CHD mortality rates in Minneapolis/St Paul fell 47% and 51% in men and women, respectively; the comparable declines in US whites were 34% and 29%. In-hospital mortality declined faster than out-of-hospital mortality. The rate of AMI (ICD-9 code 410) hospital discharges declined almost 20% between 1985 and 1995, whereas the discharge rate for unstable angina (ICD-9 code 411) increased substantially. The incidence of hospitalized definite AMI declined approximately 10%, whereas recurrence rates fell 20% to 30%. Three-year case fatality rates after hospitalized AMI decreased consistently by 31% and 41% in men and women, respectively. In-hospital administration of thrombolytic therapy, emergency angioplasty, ACE inhibitors, beta-blockers, heparin, and aspirin increased greatly. CONCLUSIONS: Declining out of-hospital death rates, declining incidence and recurrence of AMI in the population, and marked improvements in the survival of AMI patients all contributed to the 1985 to 1997 decline of CHD mortality in the Minneapolis/St Paul metropolitan area. The effects of early and late medical care seem to have had the greatest contribution to rates during this time period. PMID- 11435333 TI - Impact of viral and bacterial infectious burden on long-term prognosis in patients with coronary artery disease. AB - BACKGROUND: The number of infectious pathogens to which an individual has been exposed (infectious burden) may correlate with coronary artery disease (CAD). In a prospective study, we evaluated the effect of 8 pathogens and the aggregate pathogen burden on the risk for future fatal cardiac events among patients with angiographically documented CAD. Methods and Results-In 1018 patients, IgG or IgA antibodies to herpes simplex virus types 1 and 2, cytomegalovirus, Epstein-Barr virus, Haemophilus influenzae, Chlamydia pneumoniae, Mycoplasma pneumoniae, and Helicobacter pylori were determined. Moreover, highly sensitive C-reactive protein was measured. Follow-up information on cardiovascular events was obtained (mean 3.1 years, maximum 4.3 years). Seropositivities to Epstein-Barr virus (P=0.001), H pylori (P=0.002), and herpes simplex virus type 2 (P=0.045) were independently associated with the future risk of cardiovascular death. An increasing number for pathogen burden was significantly predictive of the long term prognosis (P<0.0001). Infectious burden divided into 0 to 3, 4 or 5, and 6 to 8 seropositivities was associated with an increasing mortality of 3.7%, 7.2%, and 12.6%, respectively. Patients seropositive to >5 pathogens compared with those seropositive to <4 pathogens had a 5.1 (1.4 to 18.3) higher risk of future cardiac death. This result was mainly driven by the pathogen burden of seropositivities to Herpesviridae (P<0.0001). The prognostic impact of total or viral pathogen burden was independent of the C-reactive protein level. CONCLUSIONS: These results support the hypothesis that the number of infectious pathogens to which an individual has been exposed independently contributes to the long-term prognosis in patients with documented CAD. PMID- 11435334 TI - Sexual dimorphism in the electrocardiographic dynamics of human ventricular repolarization: characterization in true time domain. AB - BACKGROUND: Previous characterizations of sex differences in ST-T waveform voltages have largely focused on amplitudes at selected time points during repolarization, subject to potential distortions from variations in heart rate (HR) or reliance on a JT-normalized time scale. METHODS AND RESULTS: Using digitized 12-lead ECGs from 553 normal adults (426 males) with HRs confined to 60+/-1, 70+/-1, or 80+/-1 bpm, we derived X, Y, and Z lead voltages and then generated, for each HR category by sex, summary (population mean) resultant spatial vector amplitudes (ST-T(XYZ)) and instantaneous slopes (dV/dt(XYZ)) at successive 4-ms intervals following the J point. Within each HR category, there was an early intersex divergence of ST-T(XYZ) trajectories (95% CIs nonoverlapping), with men exhibiting 2- to 3-fold greater dV/dt(XYZ) values during the ST segment and achieving greater maximum T(XYZ) and dV/dt(XYZ) values than women; descending T(XYZ) limbs were relatively more concordant between sexes but still steeper in men. The early sex differences in repolarization dynamics persisted in multiple regression analyses that took into account age and a morphometric index of left ventricular mass. In men, absolute values of extrema of T(XYZ) and dV/dt(XYZ) varied inversely with HR. CONCLUSIONS: At physiological resting HRs, the spatial ST-T vector voltage time trajectory is steeper in men than in women, beginning virtually from the J point. In addition to its mechanistic implications, the demonstration of marked sensitivity of ST-T(XYZ) and especially dV/dt(XYZ) to sex raises the possibility that these time-based, ECG-derived parameters might be informative in pathophysiological studies of ventricular repolarization. PMID- 11435335 TI - Randomized, placebo-controlled study for immunosuppressive treatment of inflammatory dilated cardiomyopathy: two-year follow-up results. AB - BACKGROUND: Previous studies have shown disappointing results for immunosuppressive treatment in patients with dilated cardiomyopathy. Therefore, we studied the effectiveness of such therapy in patients with HLA upregulation on biopsy. METHODS AND RESULTS: Of 202 patients with dilated cardiomyopathy, 84 patients with increased HLA expression were randomized to receive either immunosuppression or placebo for 3 months; they were then followed for 2 years. After 2 years, there were no significant differences in the primary end point (a composite of death, heart transplantation, and hospital readmission) between the 2 study groups (22.8% for the immunosuppression group and 20.5% for the placebo). The secondary efficacy end point included changes in ejection fraction, end diastolic diameter, end-diastolic volume, end-systolic volume and NYHA class; left ventricular ejection fraction increased significantly in the immunosuppression group compared with the placebo group (95% CI, 4.20 to 13.12; P<0.001) after 3 months of follow-up. The early favorable effects of immunosuppressive therapy on left ventricular volume, left ventricular diastolic dimension, and New York Heart Association class were also present. This improvement was maintained in the immunosuppression group at 2 years (ejection fraction: 95% CI, 6.94 to 19.04; P<0.001). In addition, on the basis of the protocol-specified definition of improvement, 71.8% patients in the immunosuppression group versus 20.9% patients in the placebo group met the criteria of improvement after 3 months (P<0.001). At the end of the follow-up period, 71.4% patients from the immunosuppression group versus 30.8% patients from the placebo group were improved (P=0.001). CONCLUSIONS: These data demonstrate a long-term benefit of immunosuppressive therapy in patients with dilated cardiomyopathy and HLA upregulation on biopsy specimens. Thus, restoration of immunosuppressive therapy for such patients should be considered. PMID- 11435336 TI - Randomized assessment of syncope trial: conventional diagnostic testing versus a prolonged monitoring strategy. AB - BACKGROUND: Establishing a diagnosis in patients with unexplained syncope is complicated by infrequent and unpredictable events. Prolonged monitoring may be an alternative strategy to conventional testing with short-term monitoring and provocative tilt and electrophysiological testing. METHODS AND RESULTS: Sixty patients (aged 66+/-14 years, 33 male) with unexplained syncope were randomized to "conventional" testing with an external loop recorder and tilt and electrophysiological testing or to prolonged monitoring with an implantable loop recorder with 1 year of monitoring. If patients remained undiagnosed after their assigned strategy, they were offered crossover to the alternate strategy. A diagnosis was obtained in 14 of 27 patients randomized to prolonged monitoring compared with 6 of 30 patients undergoing conventional testing (52% versus 20%, P=0.012). Crossover was associated with a diagnosis in 1 of 6 patients undergoing conventional testing compared with 8 of 13 patients who completed monitoring (17% versus 62%, P=0.069). Overall, prolonged monitoring was more likely to result in a diagnosis than was conventional testing (55% versus 19%, P=0.0014). Bradycardia was detected in 14 patients undergoing monitoring compared with 3 patients undergoing conventional testing (40% versus 8%, P=0.005). CONCLUSIONS: A prolonged monitoring strategy is more likely to provide a diagnosis than conventional testing in patients with unexplained syncope. Consideration should be given to earlier implementation of a monitoring strategy. PMID- 11435337 TI - Permanent cardiac pacing versus medical treatment for the prevention of recurrent vasovagal syncope: a multicenter, randomized, controlled trial. AB - BACKGROUND: This clinical investigation was performed to compare the effects of permanent dual-chamber cardiac pacing with pharmacological therapy in patients with recurrent vasovagal syncope. METHODS AND RESULTS: Patients from 14 centers were randomized to receive either a DDD pacemaker provided with rate-drop response function or the beta-blocker atenolol at the dosage of 100 mg once a day. Inclusion criteria were age >35 years, >/=3 syncopal spells in the preceding 2 years, and positive response to tilt table testing with syncope occurring in association with relative bradycardia. The primary outcome was the first recurrence of syncope after randomization. Enrollment was started in December 1997, and the first formal interim analysis was performed on July 30, 2000. By that time, 93 patients (38 men and 55 women; mean age, 58.1+/-14.3 years) had been enrolled and randomized, although follow-up data were available for all patients (46 patients in the pacemaker arm, 47 patients in the pharmacological arm). The interim analysis showed a significant effect in favor of permanent cardiac pacing (recurrence of syncope in 2 patients [4.3%] after a median of 390 days) compared with medical treatment (recurrence of syncope in 12 patients [25.5%] after a median of 135 days; OR, 0.133; 95% CI, 0.028 to 0.632; P=0.004). Consequently, enrollment and follow-up were terminated. CONCLUSIONS: DDD pacing with rate-drop response function is more effective than beta-blockade for the prevention of syncopal recurrences in highly symptomatic vasovagal fainters with relative bradycardia during tilt-induced syncope. PMID- 11435338 TI - Accurate assessment of patient effective radiation dose and associated detriment risk from radiofrequency catheter ablation procedures. AB - BACKGROUND: Radiofrequency (RF) cardiac catheter ablation procedures may require extended fluoroscopic exposure resulting in elevated radiation risk. The aim of the present study was to accurately establish RF ablation radiation risk levels and to provide means for accurate patient risk estimation from studies performed in any electrophysiology laboratory. METHODS AND RESULTS: Fluoroscopy required during cardiac ablation was classified into 4 types identified by beam orientation and irradiated tissue: (1) posteroanterior exposure during catheter advancing from the groin to the heart, (2) posteroanterior heart exposure, (3) left anterior oblique heart exposure, and (4) right anterior oblique heart exposure. The duration of each exposure was monitored in 24 patients undergoing RF cardiac ablation. Dose per minute of fluoroscopy was measured at 15 organs/tissues for each projection with the use of anthropomorphic phantom and thermoluminescence dosimetry. The effective dose rate was 219, 144, 136, and 112 mu/min for groin-to-heart posteroanterior, posteroanterior, left anterior oblique, and right anterior oblique exposure, respectively. A typical ablation procedure results in a total effective dose of 8.3 mSv per hour of fluoroscopy. The average excess of fatal cancers was estimated to be 650 and 480 per million patients undergoing RF ablation requiring 1 hour of fluoroscopy for US and UK populations, respectively. The average risk for genetic defects was determined to be 1 per million births. CONCLUSIONS: Radiation risk from RF cardiac ablation is moderate compared with other complications, but it may highly exceed radiation risk from common radiological procedures. Efforts should be made toward minimization of patient radiation risk from RF ablation procedures. PMID- 11435339 TI - C-reactive protein is an independent predictor of the rate of increase in early carotid atherosclerosis. AB - BACKGROUND: An elevated plasma concentration of high-sensitivity C-reactive protein (hs-CRP) is a strong predictor of cardiovascular events. However, there have been no longitudinal studies of the relations between development of atherosclerotic lesions and hs-CRP concentrations. Furthermore, it remains unknown whether increased hs-CRP concentrations result in the development of atherosclerosis. METHODS AND RESULTS: The study included 179 outpatients 40 to 79 years of age who were treated at our institute for traditional risk factors for cardiovascular disease. The patients had no evidence of advanced carotid atherosclerosis at the time of baseline examination. Patients underwent repeated ultrasonographic evaluation of the carotid arteries for 35+/-10 months. Blood samples were collected for hs-CRP measurements. Based on focal intima-media thickening >/=1.1 mm representing plaque, plaque number (PN) and plaque score (PS; the sum of all plaque thicknesses) were calculated. The development of atherosclerosis was estimated by the formula Deltavalue/year=(last value-baseline value)/number of follow-up years. Multivariate linear regression analysis revealed that the log-transformed value for hs-CRP concentration was not related to baseline PN or PS but was related to DeltaPN/year and DeltaPS/year (beta=0.29 and 0.30; P<0.001 for both) independently of the effect of traditional risk factors. CONCLUSIONS: During the early stages of carotid atherosclerosis, the hs CRP concentration is a marker of carotid atherosclerotic activity rather than extent of atherosclerosis. PMID- 11435340 TI - Ultrasonic echolucent carotid plaques predict future strokes. AB - BACKGROUND: We tested prospectively the hypothesis that stroke development can be predicted by echolucency of carotid atherosclerotic plaques in previously symptomatic and asymptomatic patients. METHODS AND RESULTS: We followed incidence of ipsilateral ischemic strokes for 4.4 years in 111 asymptomatic and 135 symptomatic patients with >/=50% relevant carotid artery stenosis. At inclusion, echogenicity of carotid plaques and degree of stenosis were evaluated with high resolution B-mode ultrasound with computer-assisted image processing and Doppler ultrasound, respectively. We observed 44 ipsilateral ischemic strokes. In symptomatic patients, relative risk of ipsilateral ischemic stroke for echolucent versus echorich plaques was 3.1 (95% CI, 1.3 to 7.3), whereas for 80% to 99% versus 50% to 79% stenosis, the relative risk was 1.4 (95% CI, 0.7 to 3.0). Relative to symptomatic patients with echorich 50% to 79% stenotic plaques, those with echorich 80% to 99% stenotic plaques, echolucent 50% to 79% stenotic plaques, and echolucent 80% to 99% stenotic plaques had relative risks of ipsilateral ischemic strokes of 3.1 (95%CI, 0.7 to 14), 4.2 (95% CI, 1.2 to 15), and 7.9 (95% CI, 2.1 to 30), equivalent to absolute risk increases of 11%, 18%, and 28%. This was not observed in previously asymptomatic patients. CONCLUSIONS: Echolucent plaques causing >/=50% diameter stenosis by Doppler ultrasound are associated with risk of future stroke in symptomatic but not asymptomatic individuals. This suggests that measurement of echolucency, together with degree of stenosis, may improve selection of patients for carotid endarterectomy. PMID- 11435341 TI - Dose-response study of recombinant factor VIIa/tissue factor inhibitor recombinant nematode anticoagulant protein c2 in prevention of postoperative venous thromboembolism in patients undergoing total knee replacement. AB - BACKGROUND: With the best prophylactics now available, venous thromboembolism after total knee replacement remains substantial (25% to 27%). Recombinant nematode anticoagulant protein c2 (rNAPc2) is a potent inhibitor of factor VIIa/tissue factor complex that has the potential to reduce this risk. The present study was performed to determine an efficacious and safe dose of rNAPc2 for prevention of venous thromboembolism after elective, unilateral total knee replacement. METHODS AND RESULTS: This open-label, sequential dose-ranging study was conducted in 11 centers in Canada, Europe, and the United States. Five regimens were tested. Injections were administered subcutaneously on the day of surgery (day 1) and days 3, 5, and optionally, day 7. Primary efficacy outcome was a composite of overall deep vein thrombosis based on mandatory unilateral venography (day 7+/-2) and confirmed symptomatic venous thromboembolism recorded 60 came by ambulance than those age < 12 (p << 0.01). Some 98.5% of the ambulance arrivals were emergencies; 40.7% of the ambulance arrivals were attributable to trauma versus 27.3% of the walk in arrivals. The majority of the trauma cases brought in by ambulance were because of road traffic accidents (15.3%) or home accidents (7.4%). The peak in ambulance arrivals was between 2100-2300 hours compared with 1000-1200 for the walk in arrivals. More than half of the ambulance arrivals were admitted. CONCLUSION: In planning resource allocation and in the development of contingency plans, the resource use of ambulance patients and the pattern of their arrivals should be taken into account. PMID- 11435373 TI - The effect of the introduction of NHS Direct on requests for telephone advice from an accident and emergency department. AB - OBJECTIVES: To assess the effect of the introduction of NHS Direct on advice seeking calls to an accident and emergency (A&E) department. METHODS: Review of departmental telephone advice logbook before and after the introduction of NHS Direct together with recording of the number of calls redirected to NHS Direct by the hospital switchboard and the A&E department. RESULTS: The number of advice calls responded to within the department fell by 72.6% (84 in October 1998, 23 in October 1999). The number of calls redirected to NHS Direct was 242. The total number of calls to the hospital seeking advice thus increased by 315%. CONCLUSION: The introduction of NHS Direct has allowed for a mechanism to be put in place reducing the number of calls for advice being dealt with by the A&E department clinical staff with a concomitant time saving. At the same time the number of calls for advice made by the public to the hospital has dramatically increased. PMID- 11435374 TI - An analysis of calls referred to the emergency 999 service by NHS Direct. AB - INTRODUCTION: NHS Direct was launched in West Yorkshire in April 1999. A 999 ambulance can be dispatched to the patient as a result of a call to NHS Direct. The aim of this study is to compare cases that had been referred by NHS Direct via the 999 service, with those who had dialled 999 themselves. METHODS: The study was carried out in three accident and emergency (A&E) departments in West Yorkshire, between 1 April 1999 and 21 August 1999. NHS Direct generated 999 calls were identified. The comparison group was selected at random from a list of all other 999 cases attending the three departments. The study involved retrieving basic demographic data, as well as duration of symptoms before dialling 999, triage category on arrival in A&E and disposal of patient. Ethical approval and statistical advice were obtained. RESULTS: 91 NHS Direct generated 999 calls were identified. Of the comparison group (260 cases), 28 were excluded from the study. There were no differences in the triage categories assigned to the two groups on arrival in A&E. There were notable differences in presenting complaint between the two groups and in particular, trauma was less common among the NHS Direct patients (6.6%) compared with those who had self dialled (37.5%). Patients who had first called NHS Direct were younger (p=0.033) and had endured their symptoms for longer (p<0.001); they were less likely to be admitted, and if discharged, were less likely to have follow up arranged (p=0.014). CONCLUSIONS: Both groups received similar triage categories suggesting that severity of illness is equally well assessed by self as by NHS Direct. There are large differences in case mix between the two groups studied and these almost certainly explain the differences in outcome. PMID- 11435375 TI - Dealing with emergencies in rural areas of Europe: proceedings from WONCA Europe 2000. AB - On 4 July 2000, two workshops on handling emergencies in rural areas of Europe were held at the WONCA (World Organisation of National Colleges and Academies of Family Medicine/General Practice) conference in Vienna under the auspices of EURIPA (European Rural and Isolated Practitioners' Association). Papers submitted covered varying patterns of service provision and examples of short intensive training for emergencies at resident and general practitioner levels. Ways of collecting data routinely for accident prevention were also discussed. The workshops concluded that there was a need for more research in the application of emergency skills, that lack of confidence in dealing with emergencies may contribute to recruitment problems, and that further work towards a document detailing training requirements for emergencies was needed. This will be developed at a EURIPA workshop at WONCA in Tampere, June 2001. PMID- 11435376 TI - An unusual case of subdural haematoma presenting to the accident and emergency department. AB - A case of subdural haematoma associated with an intracranial arachnoid cyst is reported. The pathogenesis, clinical presentation and treatment options of intracranial arachnoid cysts is discussed. PMID- 11435377 TI - Mild head injury with isolated third nerve palsy. AB - Traumatic isolated cranial nerve palsies are uncommon and when they do occur, they are usually associated with severe head trauma. Cranial nerve palsy associated with mild head injury is rare. A case is reported of complete left third nerve palsy associated with mild head injury. The rate of recovery for complete third nerve palsy is slow and prolonged. The ptosis recovered in 10 months; the divergent squint required botulinum toxin to the lateral rectus muscle followed by surgery. PMID- 11435378 TI - Extraluminal migration of a coin in the oesophagus of a child misdiagnosed as asthma. AB - Ingestion of a foreign body, the commonest being a coin, is a common problem in children. In most cases the coin will pass uneventfully through the gastrointestinal tract. However, on rare occasions it may become lodged in the oesophagus with subsequent extraluminal migration with the potential for serious complications such as vascular fistula or chronic suppurative infection. A case is presented of extraluminal migration of a coin in the oesophageal associated with abscess formation in a 15 month old boy. This case is particularly important because the presenting symptom of wheezing led to the erroneous diagnosis of asthma, which resulted in a three month delay in investigation and treatment. In addition, it raises the issue of whether to perform chest radiography on newly diagnosed asthmatic patients to rule out the presence of a foreign body and thereby prevent serious complications. PMID- 11435379 TI - An unusual cause of a limp in a child: developmental coxa vara. PMID- 11435380 TI - Isolated fracture of the first rib without associated injuries: a case report. PMID- 11435381 TI - An avoidable complication of digital nerve block. PMID- 11435382 TI - Tension pneumocephalus attributable to an ethmoid osteoma presenting as a stroke in evolution: an unusual presentation. PMID- 11435383 TI - Hallucal sesamoid bone stress fracture; 21st century "club foot". PMID- 11435384 TI - Tension pneumothorax: a difficult diagnosis. PMID- 11435385 TI - An unusual cause of hip pain in a child. PMID- 11435386 TI - Pain management. PMID- 11435387 TI - The immediate impact of the availability of general practitioner services on emergency department presentations. PMID- 11435390 TI - Open chest cardiac compression. PMID- 11435394 TI - Identifying functional elements by comparative DNA sequence analysis. PMID- 11435395 TI - Microsatellites in the eukaryotic DNA mismatch repair genes as modulators of evolutionary mutation rate. PMID- 11435396 TI - Systematic analysis of DNA microarray data: ordering and interpreting patterns of gene expression. PMID- 11435397 TI - The human ATP-binding cassette (ABC) transporter superfamily. AB - The ATP-binding cassette (ABC) transporter superfamily contains membrane proteins that translocate a variety of substrates across extra- and intra-cellular membranes. Genetic variation in these genes is the cause of or contributor to a wide variety of human disorders with Mendelian and complex inheritance, including cystic fibrosis, neurological disease, retinal degeneration, cholesterol and bile transport defects, anemia, and drug response. Conservation of the ATP-binding domains of these genes has allowed the identification of new members of the superfamily based on nucleotide and protein sequence homology. Phylogenetic analysis is used to divide all 48 known ABC transporters into seven distinct subfamilies of proteins. For each gene, the precise map location on human chromosomes, expression data, and localization within the superfamily has been determined. These data allow predictions to be made as to potential functions or disease phenotypes associated with each protein. In this paper, we review the current state of knowledge on all human ABC genes in inherited disease and drug resistance. In addition, the availability of the complete Drosophila genome sequence allows the comparison of the known human ABC genes with those in the fly genome. The combined data enable an evolutionary analysis of the superfamily. Complete characterization of all ABC from the human genome and from model organisms will lead to important insights into the physiology and the molecular basis of many human disorders. PMID- 11435398 TI - Conservation of microstructure between a sequenced region of the genome of rice and multiple segments of the genome of Arabidopsis thaliana. AB - The nucleotide sequence was determined for a 340-kb segment of rice chromosome 2, revealing 56 putative protein-coding genes. This represents a density of one gene per 6.1 kb, which is higher than was reported for a previously sequenced segment of the rice genome. Sixteen of the putative genes were supported by matches to ESTs. The predicted products of 29 of the putative genes showed similarity to known proteins, and a further 17 genes showed similarity only to predicted or hypothetical proteins identified in genome sequence data. The region contains a few transposable elements: one retrotransposon, and one transposon. The segment of the rice genome studied had previously been identified as representing a part of rice chromosome 2 that may be homologous to a segment of Arabidopsis chromosome 4. We confirmed the conservation of gene content and order between the two genome segments. In addition, we identified a further four segments of the Arabidopsis genome that contain conserved gene content and order. In total, 22 of the 56 genes identified in the rice genome segment were represented in this set of Arabidopsis genome segments, with at least five genes present, in conserved order, in each segment. These data are consistent with the hypothesis that the Arabidopsis genome has undergone multiple duplication events. Our results demonstrate that conservation of the genome microstructure can be identified even between monocot and dicot species. However, the frequent occurrence of duplication, and subsequent microstructure divergence, within plant genomes may necessitate the integration of subsets of genes present in multiple redundant segments to deduce evolutionary relationships and identify orthologous genes. PMID- 11435399 TI - Surveying Saccharomyces genomes to identify functional elements by comparative DNA sequence analysis. AB - Comparative sequence analysis has facilitated the discovery of protein coding genes and important functional sequences within proteins, but has been less useful for identifying functional sequence elements in nonprotein-coding DNA because the relatively rapid rate of change of nonprotein-coding sequences and the relative simplicity of non-coding regulatory sequence elements necessitates the comparison of sequences of relatively closely related species. We tested the use of comparative DNA sequence analysis to aid identification of promoter regulatory elements, nonprotein-coding RNA genes, and small protein-coding genes by surveying random DNA sequences of several Saccharomyces yeast species, with the goal of learning which species are best suited for comparisons with S. cerevisiae. We also determined the DNA sequence of a few specific promoters and RNA genes of several Saccharomyces species to determine the degree of conservation of known functional elements within the genome. Our results lead us to conclude that comparative DNA sequence analysis will enable identification of functionally conserved elements within the yeast genome, and suggest a path for obtaining this information. PMID- 11435400 TI - Phylogenetic analysis of ribonuclease H domains suggests a late, chimeric origin of LTR retrotransposable elements and retroviruses. AB - We have conducted a phylogenetic analysis of the Ribonuclease HI (RNH) domains present in Eubacteria, Eukarya, all long-term repeat (LTR)-bearing retrotransposons, and several late-branching clades of non-LTR retrotransposons. Analysis of this simple yet highly conserved enzymatic domain from these disparate sources provides surprising insights into the evolution of eukaryotic retrotransposons. First, it indicates that the lineage of elements leading to vertebrate retroviruses acquired a new RNH domain either from non-LTR retrotransposons or from a eukaryotic host genome. The preexisting retroviral RNH domain degenerated to become the tether (connection) domain of the reverse transcriptase (RT)-RNH complex. Second, it indicates that all LTR retrotransposons arose in eukaryotes well after the origin of the non-LTR retrotransposons. Because of the younger age of the LTR retrotransposons, their complex structure, and the absence of any prokaryotic precursors, we propose that the LTR retrotransposons originated as a fusion between a DNA-mediated transposon and a non-LTR retrotransposon. The resulting two-step mechanism of LTR retrotransposition, in which RNA is reverse transcribed away from the chromosomal target site, rather than directly onto the target site, was probably an adaptation to the uncoupling of transcription and translation in eukaryotic cells. PMID- 11435401 TI - Temporal gene regulation during HIV-1 infection of human CD4+ T cells. AB - CD4(+) T-cell depletion is a characteristic of human immunodeficiency virus type 1 (HIV-1) infection. In this study, modulation of mRNA expression of 6800 genes was monitored simultaneously at eight time points in a CD4(+) T-cell line (CEM GFP) during HIV infection. The responses to infection included: (1) >30% decrease at 72 h after infection in overall host-cell production of monitored mRNA synthesis, with the replacement of host-cell mRNA by viral mRNA, (2) suppression of the expression of selected mitochondrial and DNA repair gene transcripts, (3) increased expression of the proapoptotic gene and its gene p53-induced product Bax, and (4) activation of caspases 2, 3, and 9. The intense HIV-1 transcription resulted in the repression of much cellular RNA expression and was associated with the induction of apoptosis of infected cells but not bystander cells. This choreographed host gene response indicated that the subversion of the cell transcriptional machinery for the purpose of HIV-1 replication is akin to genotoxic stress and represents a major factor leading to HIV-induced apoptosis. PMID- 11435402 TI - The evolutionary chromosome translocation 4;19 in Gorilla gorilla is associated with microduplication of the chromosome fragment syntenic to sequences surrounding the human proximal CMT1A-REP. AB - Many genomic disorders occur as a result of chromosome rearrangements involving low-copy repeats (LCRs). To better understand the molecular basis of chromosome rearrangements, including translocations, we have investigated the mechanism of evolutionary rearrangements. In contrast to several intrachromosomal rearrangements, only two evolutionary translocations have been identified by cytogenetic analyses of humans and greater apes. Human chromosome 2 arose as a result of a telomeric fusion between acrocentric chromosomes, whereas chromosomes 4 and 19 in Gorilla gorilla are the products of a reciprocal translocation between ancestral chromosomes, syntenic to human chromosomes 5 and 17, respectively. Fluorescence in situ hybridization (FISH) was used to characterize the breakpoints of the latter translocation at the molecular level. We identified three BAC clones that span translocation breakpoints. One breakpoint occurred in the region syntenic to human chromosome 5q13.3, between the HMG-CoA reductase gene (HMGCR) and RAS p21 protein activator 1 gene (RASA1). The second breakpoint was in a region syntenic to human chromosome 17p12 containing the 24 kb region specific low-copy repeat-proximal CMT1A-REP. Moreover, we found that the t(4;19) is associated with a submicroscopic chromosome duplication involving a 19p chromosome fragment homologous to the human chromosome region surrounding the proximal CMT1A-REP. These observations further indicate that higher order genomic architecture involving low-copy repeats resulting from genomic duplication plays a significant role in karyotypic evolution. PMID- 11435403 TI - The repertoire of Na,K-ATPase alpha and beta subunit genes expressed in the zebrafish, Danio rerio. AB - We have identified a cohort of zebrafish expressed sequence tags encoding eight Na,K-ATPase alpha subunits and five beta subunits. Sequence comparisons and phylogenetic analysis indicate that five of the zebrafish alpha subunit genes comprise an alpha1-like gene subfamily and two are orthologs of the mammalian alpha3 subunit gene. The remaining alpha subunit clone is most similar to the mammalian alpha2 subunit. Among the five beta subunit genes, two are orthologs of the mammalian beta1 isoform, one represents a beta2 ortholog, and two are orthologous to the mammalian beta3 subunit. Using zebrafish radiation hybrid and meiotic mapping panels, we determined linkage assignments for each alpha and beta subunit gene. Na,K-ATPase genes are dispersed in the zebrafish genome with the exception of four of the alpha1-like genes, which are tightly clustered on linkage group 1. Comparative mapping studies indicate that most of the zebrafish Na,K-ATPase genes localize to regions of conserved synteny between zebrafish and humans. The expression patterns of Na,K-ATPase alpha and beta subunit genes in zebrafish are quite distinctive. No two alpha or beta subunit genes exhibit the same expression profile. Together, our data imply a very high degree of Na,K ATPase isoenzyme heterogeneity in zebrafish, with the potential for 40 structurally distinct alpha/beta subunit combinations. Differences in expression patterns of alpha and beta subunits suggest that many of the isoenzymes are also likely to exhibit differences in functional properties within specific cell and tissue types. Our studies form a framework for analyzing structure function relationships for sodium pump isoforms using reverse genetic approaches. PMID- 11435404 TI - Linkage disequilibrium between microsatellite markers extends beyond 1 cM on chromosome 20 in Finns. AB - Linkage disequilibrium (LD) is a proven tool for evaluating population structure and localizing genes for monogenic disorders. LD-based methods may also help localize genes for complex traits. We evaluated marker-marker LD using 43 microsatellite markers spanning chromosome 20 with an average density of 2.3 cM. We studied 837 individuals affected with type 2 diabetes and 386 mostly unaffected spouse controls. A test of homogeneity between the affected individuals and their spouses showed no difference, allowing the 1223 individuals to be analyzed together. Significant (P < 0.01) LD was observed using a likelihood ratio test in all (11/11) marker pairs within 1 cM, 78% (25/32) of pairs 1-3 cM apart, and 39% (7/18) of pairs 3-4 cM apart, but for only 12 of 842 pairs more than 4 cM apart. We used the human genome project working draft sequence to estimate kilobase (kb) intermarker distances, and observed highly significant LD (P < 10(-10)) for all six marker pairs up to 350 kb apart, although the correlation of LD with cM is slightly better than the correlation with megabases. These data suggest that microsatellites present at 1-cM density are sufficient to observe marker-marker LD in the Finnish population. PMID- 11435405 TI - An efficient and robust statistical modeling approach to discover differentially expressed genes using genomic expression profiles. AB - We have developed a statistical regression modeling approach to discover genes that are differentially expressed between two predefined sample groups in DNA microarray experiments. Our model is based on well-defined assumptions, uses rigorous and well-characterized statistical measures, and accounts for the heterogeneity and genomic complexity of the data. In contrast to cluster analysis, which attempts to define groups of genes and/or samples that share common overall expression profiles, our modeling approach uses known sample group membership to focus on expression profiles of individual genes in a sensitive and robust manner. Further, this approach can be used to test statistical hypotheses about gene expression. To demonstrate this methodology, we compared the expression profiles of 11 acute myeloid leukemia (AML) and 27 acute lymphoblastic leukemia (ALL) samples from a previous study (Golub et al. 1999) and found 141 genes differentially expressed between AML and ALL with a 1% significance at the genomic level. Using this modeling approach to compare different sample groups within the AML samples, we identified a group of genes whose expression profiles correlated with that of thrombopoietin and found that genes whose expression associated with AML treatment outcome lie in recurrent chromosomal locations. Our results are compared with those obtained using t-tests or Wilcoxon rank sum statistics. PMID- 11435406 TI - Predicting splice variant from DNA chip expression data. AB - Alternative splicing of premessenger RNA is an important layer of regulation in eukaryotic gene expression. Splice variation of a large number of genes has been implicated in various cell growth and differentiation processes. To measure tissue-specific splicing of genes on a large scale, we collected gene expression data from 11 rat tissues using a high-density oligonucleotide array representing 1600 rat genes. Expression of each gene on the chip is measured by 20 pairs of independent oligonucleotide probes. Two algorithms have been developed to normalize and compare the chip hybridization signals among different tissues at individual oligonucleotide probe level. Oligonucleotide probes (the perfect match [PM] probe of each probe pair), detecting potential tissue-specific splice variants, were identified by the algorithms. The identified candidate splice variants have been compared to the alternatively spliced transcripts predicted by an EST clustering program. In addition, 50% of the top candidates predicted by the algorithms were confirmed by RT-PCR experiment. The study indicates that oligonucleotide probe-based DNA chip assays provide a powerful approach to detect splice variants at genome scale. PMID- 11435407 TI - Phenotype microarrays for high-throughput phenotypic testing and assay of gene function. AB - The bacterium Escherichia coli is used as a model cellular system to test and validate a new technology called Phenotype MicroArrays (PMs). PM technology is a high-throughput technology for simultaneous testing of a large number of cellular phenotypes. It consists of preconfigured well arrays in which each well tests a different cellular phenotype and an automated instrument that continuously monitors and records the response of the cells in all wells of the arrays. For example, nearly 700 phenotypes of E. coli can be assayed by merely pipetting a cell suspension into seven microplate arrays. PMs can be used to directly assay the effects of genetic changes on cells, especially gene knock-outs. Here, we provide data on phenotypic analysis of six strains and show that we can detect expected phenotypes as well as, in some cases, unexpected phenotypes. PMID- 11435408 TI - Arrays of arrays for high-throughput gene expression profiling. AB - Gene expression profiling using DNA arrays is rapidly becoming an essential tool for research and drug discovery and may soon play a central role in disease diagnosis. Although it is possible to make significant discoveries on the basis of a relatively small number of expression profiles, the full potential of this technology is best realized through more extensive collections of expression measurements. The generation of large numbers of expression profiles can be a time-consuming and labor-intensive process with current one-at-a-time technology. We have developed the ability to obtain expression profiles in a highly parallel yet straightforward format using glass wafers that contain 49 individual high density oligonucleotide arrays. This arrays of arrays concept is generalizable and can be adapted readily to other types of arrays, including spotted cDNA microarrays. It is also scalable for use with hundreds and even thousands of smaller arrays on a single piece of glass. Using the arrays of arrays approach and parallel preparation of hybridization samples in 96-well plates, we were able to determine the patterns of gene expression in 27 ovarian carcinomas and 4 normal ovarian tissue samples, along with a number of control samples, in a single experiment. This new approach significantly increases the ease, efficiency, and throughput of microarray-based experiments and makes possible new applications of expression profiling that are currently impractical. PMID- 11435409 TI - High-throughput genotyping with single nucleotide polymorphisms. AB - To make large-scale association studies a reality, automated high-throughput methods for genotyping with single-nucleotide polymorphisms (SNPs) are needed. We describe PCR conditions that permit the use of the TaqMan or 5' nuclease allelic discrimination assay for typing large numbers of individuals with any SNP and computational methods that allow genotypes to be assigned automatically. To demonstrate the utility of these methods, we typed >1600 individuals for a G-to-T transversion that results in a glutamate-to-aspartate substitution at position 298 in the endothelial nitric oxide synthase gene, and a G/C polymorphism (newly identified in our laboratory) in intron 8 of the 11-beta hydroxylase gene. The genotyping method is accurate-we estimate an error rate of fewer than 1 in 2000 genotypes, rapid-with five 96-well PCR machines, one fluorescent reader, and no automated pipetting, over one thousand genotypes can be generated by one person in one day, and flexible-a new SNP can be tested for association in less than one week. Indeed, large-scale genotyping has been accomplished for 23 other SNPs in 13 different genes using this method. In addition, we identified three "pseudo SNPs" (WIAF1161, WIAF2566, and WIAF335) that are probably a result of duplication. PMID- 11435410 TI - High-throughput plasmid purification for capillary sequencing. AB - The need for expeditious and inexpensive methods for high-throughput DNA sequencing has been highlighted by the accelerated pace of genome DNA sequencing over the past year. At the Joint Genome Institute, the throughput in terms of high-quality bases per day has increased over 20-fold during the past 18 mo, reaching an average of 18.3 million bases per day. To support this unprecedented scaleup, we developed an inexpensive automated method for the isolation and purification of double-stranded plasmid DNA clones for sequencing that is tailored to meet the more stringent needs of the newer capillary electrophoresis DNA sequencing machines. The protocol is based on the magnetic bead method of solid phase reversible immobilization that has been automated by using a CRS based robotic system. The method described here has enabled us to meet our increases in production while reducing labor and materials costs significantly. PMID- 11435411 TI - An enhanced linkage map of the sheep genome comprising more than 1000 loci. AB - A medium-density linkage map of the ovine genome has been developed. Marker data for 550 new loci were generated and merged with the previous sheep linkage map. The new map comprises 1093 markers representing 1062 unique loci (941 anonymous loci, 121 genes) and spans 3500 cM (sex-averaged) for the autosomes and 132 cM (female) on the X chromosome. There is an average spacing of 3.4 cM between autosomal loci and 8.3 cM between highly polymorphic [polymorphic information content (PIC) > or = 0.7] autosomal loci. The largest gap between markers is 32.5 cM, and the number of gaps of > 20 cM between loci, or regions where loci are missing from chromosome ends, has been reduced from 40 in the previous map to 6. Five hundred and seventy-three of the loci can be ordered on a framework map with odds of > 1000 : 1. The sheep linkage map contains strong links to both the cattle and goat maps. Five hundred and seventy-two of the loci positioned on the sheep linkage map have also been mapped by linkage analysis in cattle, and 209 of the loci mapped on the sheep linkage map have also been placed on the goat linkage map. Inspection of ruminant linkage maps indicates that the genomic coverage by the current sheep linkage map is comparable to that of the available cattle maps. The sheep map provides a valuable resource to the international sheep, cattle, and goat gene mapping community. PMID- 11435412 TI - A sequence-based integrated map of chromosome 22. AB - The near-completion of the sequence for chromosome 22q revolutionizes map integration. We describe a sequence-based integrated map containing 968 loci including 516 known or predicted gene sequences, 317 STSs not included in these sequences, and 135 nonexpressed multinucleotide polymorphisms. The published sequence spans 34.6 Mb, inclusive of gaps estimated to total 1.1 Mb, compared with a top-down estimate of 43 Mb. This discrepancy is discussed, but will not be resolved until more of the genome is analyzed. The radiation hybrid map has 5% error in order and 34% error in location exceeding 1 Mb. The utility of a composite location based on evidence other than sequence is limited to regions not yet sequenced. A genetic map conditional on sequence order was constructed from pairwise lods. Its length of 74.8 cM in males and 80.2 cM in females is slightly less than the previous estimate not constrained by sequence order. Five recombination hot spots are detected, with differences in location between the sexes. Male recombination correlates with repetitive DNA, whereas female recombination does not. It remains to be seen whether this is true for other human chromosomes. An algorithm to improve the fit of cytogenetic bands sequence location reduces the discrepancies in cytogenetic assignment from 61 to 38. This sequence-based integrated map is represented in the genetic location database (LDB2000), which is available at http://cedar.genetics.soton.ac.uk/public_html/LDB2000.html. PMID- 11435413 TI - Massive sequence comparisons as a help in annotating genomic sequences. AB - An all-by-all comparison of all the publicly available protein sequences from plants has been performed, followed by a clusterization process. Within each of the 1064 resulting clusters-containing sequences that are orthologous as well as paralogous-the sequences have been submitted to a pyramidal classification and their domains delineated by an automated procedure a la. This process provides a means for easily checking for any apparent inconsistency in a cluster, for example, whether one sequence is shorter or longer than the others, one domain is missing, etc. In such cases, the alignment of the DNA sequence of the gene with that of a close homologous protein often reveals (in 10% of the clusters) probable sequencing errors (leading to frameshifts) or probable wrong intron/exon predictions. The composition of the clusters, their pyramidal classifications, and domain decomposition, as well as our comments when appropriate, are available from http://chlora.infobiogen.fr:1234/PHYTOPROT. PMID- 11435414 TI - Toward high-throughput genotyping: dynamic and automatic software for manipulating large-scale genotype data using fluorescently labeled dinucleotide markers. AB - To efficiently manipulate large amounts of genotype data generated with fluorescently labeled dinucleotide markers, we developed a Microsoft database management system, named. offers several advantages. First, it accommodates the dynamic nature of the accumulations of genotype data during the genotyping process; some data need to be confirmed or replaced by repeat lab procedures. By using, the raw genotype data can be imported easily and continuously and incorporated into the database during the genotyping process that may continue over an extended period of time in large projects. Second, almost all of the procedures are automatic, including autocomparison of the raw data read by different technicians from the same gel, autoadjustment among the allele fragment size data from cross-runs or cross-platforms, autobinning of alleles, and autocompilation of genotype data for suitable programs to perform inheritance check in pedigrees. Third, provides functions to track electrophoresis gel files to locate gel or sample sources for any resultant genotype data, which is extremely helpful for double-checking consistency of raw and final data and for directing repeat experiments. In addition, the user-friendly graphic interface of renders processing of large amounts of data much less labor-intensive. Furthermore, has built-in mechanisms to detect some genotyping errors and to assess the quality of genotype data that then are summarized in the statistic reports automatically generated by. The can easily handle >500,000 genotype data entries, a number more than sufficient for typical whole-genome linkage studies. The modules and programs we developed for the can be extended to other database platforms, such as Microsoft SQL server, if the capability to handle still greater quantities of genotype data simultaneously is desired. PMID- 11435415 TI - A new member of the family of di-iron carboxylate proteins. Coq7 (clk-1), a membrane-bound hydroxylase involved in ubiquinone biosynthesis. AB - Ubiquinone (UQ) is an essential cofactor for respiratory metabolism. In yeast, mutation of the COQ7 gene results in the absence of UQ biosynthesis and demonstrates a role for this gene in the step leading to the hydroxylation of 5 demethoxyubiquinone. Intriguingly, the disruption of the corresponding gene in Caenorhabditis elegans, clk-1, results in a prolonged life span and a slowing of development. Because of the pleiotropic effect of this disruption, the small size of the protein, and the lack of obvious homology to other known hydroxylases, it has been suggested that Coq7 may be a regulatory or structural component in UQ biosynthesis, rather than acting as the hydroxylase per se. Here we identify Coq7 as belonging to a family of a di-iron containing oxidases/hydroxylases based on a conserved sequence motif for the iron ligands, supporting a direct function of Coq7 as a hydroxylase. We have cloned COQ7 from Pseudomonas aeruginosa and Thiobacillus ferrooxidans and show that indeed this gene complements an Escherichia coli mutant that lacks an unrelated 5-demethoxyubiquinone hydroxylase. Based on the similarities to other well studied di-iron carboxylate proteins, we propose a structural model for Coq7 as an interfacial integral membrane protein. PMID- 11435416 TI - Calcineurin-GATA4 pathway is involved in beta-adrenergic agonist-responsive endothelin-1 transcription in cardiac myocytes. AB - Increases in the expression of endothelin-1 (ET-1) in cardiac myocytes play a critical role in the development of heart failure in vivo. Whereas norepinephrine (NE) is a potent inducer of ET-1 expression in cardiac myocytes, the signaling pathways that link NE to inducible cardiac ET-1 expression are unknown. Adrenergic stimulation results in an increase in intracellular calcium levels, which in turn activates calcineurin. Here, we have shown that stimulation with NE markedly increased the expression of the ET-1 gene in primary cardiac myocytes from neonatal rats. This increase was severely attenuated by a beta-adrenergic antagonist, metoprolol, but not by an alpha-adrenergic antagonist, prazosin. Consistent with these data, the beta-adrenergic agonist isoproterenol (ISO) activated the rat ET-1 promoter activity to an extent that was similar to NE. The ISO-stimulated increase in promoter activity was significantly inhibited by a Ca(2+)-antagonist, nifedipine, and an immunosuppressant, cyclosporin A, which blocks calcineurin. Mutation analysis indicated that the GATA4 binding site is required for ISO-responsive ET-1 transcription. Stimulation with ISO enhanced the interaction between NFATc and GATA4 in cardiac myocytes. Consistent with this interaction, overexpression of GATA4 and NFATc synergistically activated the ET-1 promoter. These findings demonstrate that NE-stimulated ET-1 expression in cardiac myocytes is mediated predominantly via a beta-adrenergic pathway, and that calcium-activated calcineurin-GATA4 plays a role in this process. PMID- 11435417 TI - The cytoplasmic and transmembrane domains of the p75 and Trk A receptors regulate high affinity binding to nerve growth factor. AB - Ligand-induced receptor oligomerization is an established mechanism for receptor tyrosine kinase activation. However, numerous receptor-tyrosine kinases are expressed in multicomponent complexes with other receptors that may signal independently or alter the binding characteristics of the receptor-tyrosine kinase. Nerve growth factor (NGF) interacts with two structurally unrelated receptors, the Trk A receptor-tyrosine kinase and p75, a tumor necrosis factor receptor family member. Each receptor binds independently to NGF with predominantly low affinity (K(d) = 10(-9) m), but they produce high affinity binding sites (K(d) = 10(-11) m) upon receptor co-expression. Here we provide evidence that the number of high affinity sites is regulated by the ratio of the two receptors and by specific domains of Trk A and p75. Co-expression of Trk A containing mutant transmembrane or cytoplasmic domains with p75 yielded reduced numbers of high affinity binding sites. Similarly, co-expression of mutant p75 containing altered transmembrane and cytoplasmic domains with Trk A also resulted in predominantly low affinity binding sites. Surprisingly, extracellular domain mutations of p75 that abolished NGF binding still generated high affinity binding with Trk A. These results indicate that the transmembrane and cytoplasmic domains of Trk A and p75 are responsible for high affinity site formation and suggest that p75 alters the conformation of Trk A to generate high affinity NGF binding. PMID- 11435418 TI - Interaction of lecithin:cholesterol acyltransferase (LCAT).alpha 2-macroglobulin complex with low density lipoprotein receptor-related protein (LRP). Evidence for an alpha 2-macroglobulin/LRP receptor-mediated system participating in LCAT clearance. AB - The reaction of lecithin:cholesterol acyltransferase (LCAT) with high density lipoproteins (HDL) is of critical importance in reverse cholesterol transport, but the structural and functional pathways involved in the regulation of LCAT have not been established. We present evidence for the direct binding of LCAT to alpha(2)-macroglobulin (alpha(2)M) in human plasma to form a complex 18.5 nm in diameter. Forty percent of plasma LCAT-HDL was associated with alpha(2)M; moreover, most of the LCAT in cerebrospinal fluid and in the medium of cultured human hepatoma cell line was associated with alpha(2)M. Purified recombinant human LCAT (rLCAT) labeled with (125)I bound to native and methylamine-activated alpha(2)M (alpha(2)M-MA) in vitro in a time- and concentration-dependent manner, and this binding did not depend on the presence of lipid. rLCAT bound to alpha(2)M-MA with greater affinity than to alpha(2)M. Furthermore, rLCAT did not activate alpha(2)M as phosphatidylcholine-specific phospholipase C does. Reconstituted HDL particles (LpA-I) inhibited the binding of rLCAT to alpha(2)M more efficiently than native HDL(3) did. LCAT associated with alpha(2)M was enzymatically inactive under both endogenous and exogenous assay conditions. Purified rLCAT alone did not bind to low density lipoprotein receptor-related protein (LRP) as lipoprotein lipase (LPL) does; however, when rLCAT was combined with alpha(2)M-MA to form a complex, binding, internalization, and degradation of rLCAT took place in LRP-expressing cells (LRP (+/+)) but not in cells deficient in LRP (LRP (-/-)). It is concluded that the binding of LCAT to alpha(2)M inhibits its enzymatic activity. Furthermore, the finding supports the possibility that the LRP receptor can act in vivo to mediate clearance of the LCAT-alpha(2)M complex and may significantly influence the bioavailability of LCAT. PMID- 11435419 TI - PYK2 links G(q)alpha and G(13)alpha signaling to NF-kappa B activation. AB - Signaling via a variety of G-protein-coupled receptors (GPCRs) leads to activation of nuclear factor (NF)-kappa B. Evidence exists for a signaling pathway initiated by the B2 type bradykinin receptor via G(q) activation, which leads to the sequential stimulation of phosphoinositide 3-kinase (PI3K), the serine/threonine kinase Akt, I kappa B kinases, and finally nuclear factor NF kappa B-dependent transcription. GPCR-mediated G(q)alpha or G(13)alpha activation also potently stimulates the tyrosine kinase PYK2. In this study we tested whether G(q)alpha- and/or G(13)alpha-induced PYK2 activation contributes to GPCR mediated NF-kappa B activation. Among the GTPase-deficient forms of G alpha tested, G(13)alpha and G(q)alpha most potently stimulated an NF-kappa B-dependent reporter gene. PYK2 activated the same reporter gene and synergized with either G(q)alpha Q209L (QL) or G(13)alpha Q226L (QL). Placing PYK2 upstream of both PI3K and Akt activation, PYK2 activated Akt through a PI3K-dependent pathway, and either a dominant negative form of Akt or the PI3K inhibitor LY294002 blocked PYK2-stimulated NF-kappa B-dependent transcription. Placing PYK2 downstream of G protein activation, a kinase-dead form of PYK2, PYK2 (KD), blocked NF-kappa B dependent transcription triggered by signaling through the muscarinic receptor type 1 and either G(q)alpha QL or G(13)alpha QL. PYK2 (KD) also blocked Akt activation by the same stimuli. These results indicate that PYK2 can link G protein activation through PI3K, Akt, and I kappa B kinase to NF-kappa B activation. PMID- 11435420 TI - Interfacial positioning and stability of transmembrane peptides in lipid bilayers studied by combining hydrogen/deuterium exchange and mass spectrometry. AB - Nano-electrospray ionization mass spectrometry (ESI-MS) was used to analyze hydrogen/deuterium (H/D) exchange properties of transmembrane peptides with varying length and composition. Synthetic transmembrane peptides were used with a general acetyl-GW(2)(LA)(n)LW(2)A-ethanolamine sequence. These peptides were incorporated in large unilamellar vesicles of 1,2-dimyristoyl-sn-glycero-3 phosphocholine. The vesicles were diluted in buffered deuterium oxide, and the H/D exchange after different incubation times was directly analyzed by means of ESI-MS. First, the influence of the length of the hydrophobic Leu-Ala sequence on exchange behavior was investigated. It was shown that longer peptide analogs are more protected from H/D exchange than expected on the basis of their length with respect to bilayer thickness. This is explained by an increased protection from the bilayer environment, because of stretching of the lipid acyl chains and/or tilting of the longer peptides. Next, the role of the flanking tryptophan residues was investigated. The length of the transmembrane part that shows very slow H/D exchange was found to depend on the exact position of the tryptophans in the peptide sequence, suggesting that tryptophan acts as a strong determinant for positioning of proteins at the membrane/water interface. Finally, the influence of putative helix breakers was studied. It was shown that the presence of Pro in the transmembrane segment results in much higher exchange rates as compared with Gly or Leu, suggesting a destabilization of the alpha-helix. Tandem MS measurements suggested that the increased exchange takes place over the entire transmembrane segment. The results show that ESI-MS is a convenient technique to gain detailed insight into properties of peptides in lipid bilayers by monitoring H/D exchange kinetics. PMID- 11435421 TI - TcUBP-1, a developmentally regulated U-rich RNA-binding protein involved in selective mRNA destabilization in trypanosomes. AB - Developmental stages of the trypanosome life cycle differ in their morphology, biology, and biochemical properties. Consequently, several proteins have to be tightly regulated in their expression to allow trypanosomes to adapt rapidly to sudden environmental changes, a process that might be of central importance for parasite survival. However, in contrast to higher eukaryotic cells, trypanosomes do not seem to regulate gene expression through regulation of transcription initiation. These parasites make use of post-transcriptional regulatory mechanisms and modification of mRNA half-life is a relevant one. Trans-acting factors binding to cis-elements that affect mRNA stability of mature transcripts have not been identified in these cells. In this work, a novel U-rich RNA-binding protein (TcUBP-1) from Trypanosoma cruzi, the agent of Chagas disease, was identified. Its structure includes an RNA recognition motif, a nuclear export signal, and auxiliary domains with glycine- and glutamine-rich regions. TcUBP-1 recognizes the 44-nucleotide AU-rich RNA instability element located in the 3' untranslated region of mucin SMUG mRNAs (Di Noia, J. M., D'Orso, I., Sanchez, D. O., and Frasch, A. C. (2000) J. Biol. Chem. 275, 10218-10227) as well as GU-rich sequences. Over-expression of TcUBP-1 in trypanosomes decreases the half-life of SMUG mucin mRNAs in vivo but does not affect the stability of other parasite mRNAs. Because TcUBP-1 is developmentally regulated, it might have a relevant role in regulating protein expression during trypanosome differentiation, allowing a correct expression pattern of U-rich-containing mRNAs. PMID- 11435422 TI - Photochemical reaction cycle and proton transfers in Neurospora rhodopsin. AB - It was recently found that NOP-1, a membrane protein of Neurospora crassa, shows homology to haloarchaeal rhodopsins and binds retinal after heterologous expression in Pichia pastoris. We report on spectroscopic properties of the Neurospora rhodopsin (NR). The photocycle was studied with flash photolysis and time-resolved Fourier-transform infrared spectroscopy in the pH range 5-8. Proton release and uptake during the photocycle were monitored with the pH-sensitive dye, pyranine. Kinetic and spectral analysis revealed six distinct states in the NR photocycle, and we describe their spectral properties and pH-dependent kinetics in the visible and infrared ranges. The phenotypes of the mutant NR proteins, D131E and E142Q, in which the homologues of the key carboxylic acids of the light-driven proton pump bacteriorhodopsin, Asp-85 and Asp-96, were replaced, show that Glu-142 is not involved in reprotonation of the Schiff base but Asp-131 may be. This implies that, if the NR photocycle is associated with proton transport, it has a low efficiency, similar to that of haloarchaeal sensory rhodopsin II. Fourier-transform Raman spectroscopy revealed unexpected differences between NR and bacteriorhodopsin in the configuration of the retinal chromophore, which may contribute to the less effective reprotonation switch of NR. PMID- 11435423 TI - U box proteins as a new family of ubiquitin-protein ligases. AB - The U box is a domain of approximately 70 amino acids that is present in proteins from yeast to humans. The prototype U box protein, yeast Ufd2, was identified as a ubiquitin chain assembly factor that cooperates with a ubiquitin-activating enzyme (E1), a ubiquitin-conjugating enzyme (E2), and a ubiquitin-protein ligase (E3) to catalyze ubiquitin chain formation on artificial substrates. E3 enzymes are thought to determine the substrate specificity of ubiquitination and have been classified into two families, the HECT and RING finger families. Six mammalian U box proteins have now been shown to mediate polyubiquitination in the presence of E1 and E2 and in the absence of E3. These U box proteins exhibited different specificities for E2 enzymes in this reaction. Deletion of the U box or mutation of conserved amino acids within it abolished ubiquitination activity. Some U box proteins catalyzed polyubiquitination by targeting lysine residues of ubiquitin other than lysine 48, which is utilized by HECT and RING finger E3 enzymes for polyubiquitination that serves as a signal for proteolysis by the 26 S proteasome. These data suggest that U box proteins constitute a third family of E3 enzymes and that E4 activity may reflect a specialized type of E3 activity. PMID- 11435424 TI - The amber codon in the gene encoding the monomethylamine methyltransferase isolated from Methanosarcina barkeri is translated as a sense codon. AB - Each of the genes encoding the methyltransferases initiating methanogenesis from trimethylamine, dimethylamine, or monomethylamine by various Methanosarcina species possesses one naturally occurring in-frame amber codon that does not appear to act as a translation stop during synthesis of the biochemically characterized methyltransferase. To investigate the means by which suppression of the amber codon within these genes occurs, MtmB, a methyltransferase initiating metabolism of monomethylamine, was examined. The C-terminal sequence of MtmB indicated that synthesis of this mtmB1 gene product did not cease at the internal amber codon, but at the following ochre codon. Antibody raised against MtmB revealed that Escherichia coli transformed with mtmB1 produced the amber termination product. The same antibody detected primarily a 50-kDa protein in Methanosarcina barkeri, which is the mass predicted for the amber readthrough product of the mtmB1 gene. Sequencing of peptide fragments from MtmB by Edman degradation and mass spectrometry revealed no change in the reading frame during mtmB1 expression. The amber codon position corresponded to a lysyl residue using either sequencing technique. The amber codon is thus read through during translation at apparently high efficiency and corresponds to lysine in tryptic fragments of MtmB even though canonical lysine codon usage is encountered in other Methanosarcina genes. PMID- 11435425 TI - TEL-JAK2 mediates constitutive activation of the phosphatidylinositol 3' kinase/protein kinase B signaling pathway. AB - A subset of chromosomal translocations that participate in leukemia involve activated tyrosine kinases. The ets transcription factor, TEL, undergoes translocations with several distinct tyrosine kinases including JAK2. TEL-JAK2 transforms cell lines to factor independence, and constitutive tyrosine kinase activity results in the phosphorylation of several substrates including STAT1, STAT3, and STAT5. In this study we have shown that TEL-JAK2 can constitutively activate the phosphatidylinositol 3'-kinase (PI 3'-kinase) signaling pathway. The regulatory subunit of PI 3'-kinase, p85, associates with TEL-JAK2 in immunoprecipitations, and this was shown to be mediated by the amino-terminal SH2 domain of p85 but independent of a putative p85-binding motif within TEL-JAK2. The scaffolding protein Gab2 can also mediate the association of p85. TEL-JAK2 constitutively phosphorylates the downstream substrate protein kinase B/AKT. Importantly, the pharmacologic PI 3'-kinase inhibitor, LY294002, blocked TEL-JAK2 factor-independent growth and phosphorylation of protein kinase B. However, LY294002 did not alter STAT5 tyrosine phosphorylation, indicating that STAT5 and protein kinase B activation mediated by TEL-JAK2 are independent signaling pathways. Therefore, activation of the PI 3'-kinase signaling pathway is an important event mediated by TEL-JAK2 chromosomal translocations. PMID- 11435426 TI - Tyrosine residues 951 and 1059 of vascular endothelial growth factor receptor-2 (KDR) are essential for vascular permeability factor/vascular endothelial growth factor-induced endothelium migration and proliferation, respectively. AB - Vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) exerts its multiple functions by activating two receptor tyrosine kinases, Flt-1 (VEGFR 1) and KDR (VEGFR-2), both of which are selectively expressed on primary vascular endothelium. To dissect the respective signaling pathways and biological functions mediated by these receptors in primary endothelial cells with two receptors intact, we, recently developed chimeric receptors (EGDR and EGLT) in which the extracellular domain of the epidermal growth factor receptor was fused to the transmembrane domain and intracellular domain of KDR and Flt-1, respectively. With these fusion receptors, we have shown that KDR is solely responsible for VPF/VEGF-induced human umbilical vein endothelial cell (HUVEC) proliferation and migration, whereas Flt-1 showed an inhibitory effect on KDR mediated proliferation but not migration. To further characterize the VPF/VEGF stimulated HUVEC proliferation and migration here, we have created several EGDR mutants by site-directed mutagenesis. We show that tyrosine residues 1059 and 951 of KDR are essential for VPF/VEGF-induced HUVEC proliferation and migration, respectively. Furthermore, the mutation of tyrosine 1059 to phenylanaline results in the complete loss of KDR/EGDR-mediated intracellular Ca(2+) mobilization and MAPK phosphorylation, but the mutation of tyrosine 951 to phenylanaline did not affect these events. Our results suggest that KDR mediates different signaling pathways for HUVEC proliferation and migration and, moreover, intracellular Ca(2+) mobilization and MAPK phosphorylation are not essential for VPF/VEGF induced HUVEC migration. PMID- 11435427 TI - Cathepsin B, L, and S cleave and inactivate secretory leucoprotease inhibitor. AB - A number of serine proteases, matrix metalloproteases, and cysteine proteases were evaluated for their ability to cleave and inactivate the antiprotease, secretory leucoprotease inhibitor (SLPI). None of the serine proteases or the matrix metalloproteases examined cleaved the SLPI protein. However, incubation with cathepsins B, L, and S resulted in the cleavage and inactivation of SLPI. All three cathepsins initially cleaved SLPI between residues Thr(67) and Tyr(68). The proteolytic cleavage of SLPI by all three cathepsins resulted in the loss of the active site of SLPI and the inactivation of SLPI anti-neutrophil elastase capacity. Cleavage and inactivation were catalytic with respect to the cathepsins, so that the majority of a 400-fold excess of SLPI was inactivated within 15 min by cathepsins L and S. Analysis of epithelial lining fluid samples from individuals with emphysema indicated the presence of cleaved SLPI in these samples whereas only intact SLPI was observed in control epithelial lining fluid samples. Active cathepsin L was shown to be present in emphysema epithelial lining fluid and inhibition of this protease prevented the cleavage of recombinant SLPI added to emphysema epithelial lining fluid. Taken together with previous data that demonstrates that cathepsin L inactivates alpha(1) antitrypsin, these findings indicate the involvement of cathepsins in the diminution of the lung antiprotease screen possibly leading to lung destruction in emphysema. PMID- 11435428 TI - Transcriptional regulation of the murine acetyl-CoA synthetase 1 gene through multiple clustered binding sites for sterol regulatory element-binding proteins and a single neighboring site for Sp1. AB - Cytosolic acetyl-CoA synthetase (AceCS1) activates acetate to supply the cells with acetyl-CoA for lipid synthesis. The cDNA for the mammalian AceCS1 has been isolated recently, and the mRNA was shown to be negatively regulated by sterols in cultured cells. In the current study, we describe the molecular mechanisms directing the sterol-regulated expression of murine AceCS1 by cloning and functional studies of the 5'-flanking region of the AceCS1 gene. An AceCS1 promoter-reporter gene (approximately 2.1 kilobase pairs) was negatively regulated when sterols were added to the medium of cultured cells, and the promoter was markedly induced by co-transfection of a plasmid that expresses the transcriptionally active nuclear form of either sterol regulatory element-binding protein (SREBP)-1a or -2 in HepG2 cells. Sequence analysis suggested that the AceCS1 promoter contains an E-box, two putative CCAAT-boxes, eight sterol regulatory element (SRE) motifs, and six GC-boxes. Gel shift assays demonstrated that all eight SRE motifs bound purified SREBP-1a in vitro with similar affinity. Luciferase reporter gene assays revealed that sterol regulation was critically dependent on three closely spaced SRE motifs and an adjacent GC-box. However, mutation of two putative upstream CCAAT-boxes did not affect SREBP dependent activation. Electrophoretic mobility "supershift" analyses confirmed that both Sp1 and Sp3 bound to the critical GC-box. In addition, transfection studies in Drosophila SL2 cells demonstrated that SREBP synergistically activated the AceCS1 promoter along with Sp1 or Sp3 but not with nuclear factor-Y. PMID- 11435429 TI - Bisphosphonates are potent inhibitors of Trypanosoma cruzi farnesyl pyrophosphate synthase. AB - We report the cloning and sequencing of a gene encoding the farnesyl pyrophosphate synthase of Trypanosoma cruzi. The protein (T. cruzi farnesyl pyrophosphate synthase, TcFPPS) is an attractive target for drug development, since the growth of T. cruzi is inhibited by carbocation transition state/reactive intermediate analogs of its substrates, the nitrogen-containing bisphosphonates currently in use in bone resorption therapy. The protein predicted from the nucleotide sequence of the gene has 362 amino acids and a molecular mass of 41.2 kDa. Several sequence motifs found in other FPPSs are present in TcFPPS. Heterologous expression of TcFPPS in Escherichia coli produced a functional enzyme that was inhibited by the nitrogen-containing bisphosphonates alendronate, pamidronate, homorisedronate, and risedronate but was less sensitive to the non-nitrogen-containing bisphosphonate etidronate, which, unlike the nitrogen-containing bisphosphonates, does not affect parasite growth. The protein contains a unique 11-mer insertion located near the active site, together with other sequence differences that may facilitate the development of novel anti Chagasic agents. PMID- 11435430 TI - Inhibitory specificity and potency of proSAAS-derived peptides toward proprotein convertase 1. AB - Prohormone convertase 1 (PC1), mediating the proteolytic processing of neural and endocrine precursors, is thought to be regulated by the neuroendocrine protein proSAAS. The PC1 inhibitory sequence is mostly confined within a 10-12-amino acid segment near the C terminus of the conserved human proSAAS and contains the critical KR(244) dibasic motif. Our results show that the decapeptide proSAAS (235-244)( 235)VLGALLRVKR(244) is the most potent reversible competitive PC1 inhibitor (K(i) approximately 9 nm). The C-terminally extended proSAAS-(235-246) exhibits a 5-6-fold higher K(i) ( approximately 51 nm). The additional LE sequence at P1'-P2', resulted in a competitive substrate cleaved by PC1 at KR(244) downward arrowLE(246). Systematic alanine scanning and in some cases lysine scanning tested the contribution of each residue within proSAAS-(235-246) toward the PC1-inhibition's specificity and potency. The amino acids P1 Arg, P2 Lys, and P4 Arg are all critical for inhibition. Moreover, the aliphatic P3 Val and P5, P6, and P1' Leu significantly affect the degree of enzyme inactivation and PC1 specificity. Interestingly, a much longer N- and C-terminally extended endogenous rat proSAAS-(221-254) called little PenLen, was found to be a 3-fold less potent PC1 inhibitor with reduced selectivity but a much better substrate than proSAAS-(235-246). Molecular modeling studies and circular dichroism analysis indicate an extended and poly-l-proline II type structural conformation for proSAAS-(235-244), the most potent PC1 inhibitor, a feature not present in poor PC1 inhibitors. PMID- 11435431 TI - p190RhoGEF Binds to a destabilizing element in the 3' untranslated region of light neurofilament subunit mRNA and alters the stability of the transcript. AB - Stabilization of neurofilament (NF) mRNAs plays a major role in regulating levels of NF expression and in establishing axonal size and rate of axonal conduction. Previous studies have identified a 68-nucleotide destabilizing element at the junction of the coding region and 3' untranslated region of the light NF subunit (NF-L) mRNA. The present study has used the destabilizing element (probe A) to screen a rat brain cDNA library for interactive proteins. A cDNA clone encoding 1068 nucleotides in the C-terminal domain of p190RhoGEF (clone 39) was found to bind strongly and specifically to the RNA probe. The interaction was confirmed using a glutathione S-transferase/clone 39 fusion protein in Northwestern, gel shift, and cross-linkage studies. The glutathione S-transferase/clone 39 fusion protein also enhanced the cross-linkage of a major 43-kDa protein in brain extract to the destabilizing element. Functional studies on stably transfected neuronal cells showed that p190RhoGEF expression increased the half-life of a wild-type NF-L mRNA but did not alter the half-life of a mutant NF-L mRNA lacking the destabilizing element. The findings reveal a novel interactive feature of p190RhoGEF that links the exchange factor with NF mRNA stability and regulation of the axonal cytoskeleton. PMID- 11435432 TI - Functional differences between the human ATP-dependent nucleosome remodeling proteins BRG1 and SNF2H. AB - ATP-dependent nucleosome remodeling complexes can be grouped into several classes that may differ in their biochemical remodeling activities and biological roles. Although there are a number of biochemical studies of each class of remodeler, there are very little data directly comparing the biochemical activities of remodelers from different classes. We have purified two ATP-hydrolyzing proteins, SNF2H and BRG1, which are members of complexes from two different classes of remodelers. Consistent with previous reports, these two homogeneous proteins can perform remodeling functions. We show significant functional differences between SNF2H and BRG1 in vitro; although both SNF2H and BRG1 hydrolyze ATP and remodel linear arrays of nucleosomes, only BRG1 can remodel mononucleosomes. Also, only BRG1 can alter the topology of nucleosomal plasmids. We propose that these functional differences reflect significant mechanistic differences between the two remodeler classes that will impact their biological roles. PMID- 11435433 TI - Electron spin resonance and fluorescence studies of the bound-state conformation of a model protein substrate to the chaperone SecB. AB - SecB is a homotetrameric, cytosolic chaperone that forms part of the protein translocation machinery in Escherichia coli. We have investigated the bound-state conformation of a model protein substrate of SecB, bovine pancreatic trypsin inhibitor (BPTI) as well as the conformation of SecB itself by using proximity relationships based on site-directed spin-labeling and pyrene fluorescence methods. BPTI is a 58-residue protein and contains three disulfide groups between residues 5 and 55, 14 and 38, as well as 30 and 51. Mutants of BPTI that contained only a single disulfide were reduced, and the free cysteines were labeled with either thiol-specific spin labels or pyrene maleimide. The relative proximity of the labeled residues was studied using either electron spin resonance spectroscopy or fluorescence spectroscopy. The data suggest that SecB binds a collapsed coil of reduced unfolded BPTI, which then undergoes a structural rearrangement to a more extended state upon binding to SecB. Binding occurs at multiple sites on the substrate, and the binding site on each SecB monomer accommodates less than 21 substrate residues. In addition, we have labeled four solvent-accessible cysteine residues in the SecB tetramer and have investigated their relative spatial arrangement in the presence and absence of the substrate protein. The electron spin resonance data suggest that these cysteine residues are in close proximity (15 A) when no substrate protein is bound but move away to a distance of greater than 20 A when SecB binds substrate. This is the first direct evidence of a conformational change in SecB upon binding of a substrate protein. PMID- 11435434 TI - Targeting of EBNA1 for rapid intracellular degradation overrides the inhibitory effects of the Gly-Ala repeat domain and restores CD8+ T cell recognition. AB - Epstein-Barr virus (EBV)-encoded nuclear antigen 1 (EBNA1) includes a unique glycine-alanine repeat domain that inhibits the endogenous presentation of cytotoxic T lymphocyte (CTL) epitopes through the class I pathway by blocking proteasome-dependent degradation of this antigen. This immune evasion mechanism has been implicated in the pathogenesis of EBV-associated diseases. Here, we show that cotranslational ubiquitination combined with N-end rule targeting enhances the intracellular degradation of EBNA1, thus resulting in a dramatic reduction in the half-life of the antigen. Using DNA expression vectors encoding different forms of ubiquitinated EBNA1 for in vivo studies revealed that this rapid degradation, remarkably, leads to induction of a very strong CTL response to an EBNA1-specific CTL epitope. Furthermore, this targeting also restored the endogenous processing of HLA class I-restricted CTL epitopes within EBNA1 for immune recognition by human EBV-specific CTLs. These observations provide, for the first time, evidence that the glycine-alanine repeat-mediated proteasomal block on EBNA1 can be reversed by specifically targeting this antigen for rapid degradation resulting in enhanced CD8+ T cell-mediated recognition in vitro and in vivo. PMID- 11435435 TI - Human homeobox HOXA7 regulates keratinocyte transglutaminase type 1 and inhibits differentiation. AB - Keratinocyte proliferation and differentiation result from expression of specific groups of genes regulated by unique combinations of transcription factors. To better understand these regulatory processes, we studied HOXA7 expression and its regulation of differentiation-specific keratinocyte genes. We isolated the homeobox transcription factor HOXA7 from keratinocytes through binding to a differentiation-dependent viral enhancer and analyzed its effect on endogenous differentiation-dependent genes, primarily transglutaminase 1. HOXA7 overexpression repressed transglutaminase 1-reporter activity. HOXA7 message markedly decreased, and transglutaminase RNA increased, upon phorbol ester induced differentiation, in a protein kinase C-dependent manner. Overexpression of HOXA7 attenuated the transglutaminase 1 induction by phorbol ester, demonstrating that HOXA7 expression is inversely related to keratinocyte differentiation, and to transglutaminase 1 expression. Antisense HOXA7 expression activated transglutaminase 1, involucrin, and keratin 10 message and protein levels, demonstrating that endogenous HOXA7 down-regulates multiple differentiation-specific keratinocyte genes. In keeping with these observations, epidermal growth factor receptor activation stimulated HOXA7 expression. HOX genes function in groups, and we found that HOXA5 and HOXB7 were also down regulated by phorbol ester. These results provide the first example of protein kinase C-mediated homeobox gene regulation in keratinocytes, and new evidence that HOXA7, potentially in conjunction with HOXA5 and HOXAB7, silences differentiation-specific genes during keratinocyte proliferation, that are then released from inhibition in response to differentiation signals. PMID- 11435436 TI - Identification of novel HLA-B27 ligands derived from polymorphic regions of its own or other class I molecules based on direct generation by 20 S proteasome. AB - HLA-B27 is strongly associated with ankylosing spondylitis. Natural HLA-B27 ligands derived from polymorphic regions of its own or other class I HLA molecules might be involved in autoimmunity or provide diversity among HLA-B27 bound peptide repertoires from individuals. In particular, an 11-mer spanning HLA B27 residues 169-179 is a natural HLA-B27 ligand with homology to proteins from Gram-negative bacteria. Proteasomal digestion of synthetic substrates demonstrated direct generation of the B27-(169-179) ligand. Cleavage after residue 181 generated a B27-(169-181) 13-mer that was subsequently found as a natural ligand of B*2705 and B*2704. Its binding to HLA-B27 subtypes in vivo correlated better than B27-(169-179) with association to spondyloarthropathy. Proteasomal cleavage generated also a peptide spanning B*2705 residues 150-158. This region is polymorphic among HLA-B27 subtypes and class I HLA antigens. The peptide was a natural B*2704 ligand. Since this subtype differs from B*2705 at residue 152, it was concluded that the ligand arose from HLA-B*3503, synthesized in the cells used as a source for B*2704-bound peptides. Thus, polymorphic HLA B27 ligands derived from HLA-B27 or other class I molecules are directly produced by the 20 S proteasome in vitro, and this can be used for identification of such ligands in the constitutive HLA-B27-bound peptide pool. PMID- 11435437 TI - Barley lipid transfer protein, LTP1, contains a new type of lipid-like post translational modification. AB - In plants a group of proteins termed nonspecific lipid transfer proteins are found. These proteins bind and catalyze transfer of lipids in vitro, but their in vivo function is unknown. They have been suggested to be involved in different aspects of plant physiology and cell biology, including the formation of cutin and involvement in stress and pathogen responses, but there is yet no direct demonstration of an in vivo function. We have found and characterized a novel post-translational modification of the barley nonspecific lipid transfer protein, LTP1. The protein-modification bond is of a new type in which an aspartic acid in LTP1 is bound to the modification through what most likely is an ester bond. The chemical structure of the modification has been characterized by means of two dimensional homo- and heteronuclear nuclear magnetic resonance spectroscopy as well as mass spectrometry and is found to be lipid-like in nature. The modification does not resemble any standard lipid post-translational modification but is similar to a compound with known antimicrobial activity. PMID- 11435438 TI - Dynamically acetylated histone association with transcriptionally active and competent genes in the avian adult beta-globin gene domain. AB - In chicken immature erythrocytes, class 1 acetylated histones are rapidly tri- and tetra-acetylated and rapidly deacetylated. Class 2 acetylated H3 and H4 are rapidly acetylated to mono- and di-acetylated isoforms and slowly deacetylated. Our previous studies suggested that class 1 acetylated histones were primarily associated with transcriptionally active DNA (beta(A)-globin) but not competent DNA (epsilon-globin). Chromatin salt solubility (chromatin fiber oligomerization) is directly influenced by hyperacetylation. In this study we investigated the association of class 1 histones with beta(A)- and epsilon-globin DNA by measuring their loss of solubility rates in 150 mm NaCl and 3 mm MgCl(2) as a function of hyperacetylated histone deacetylation. Expressed and competent chromatin was associated with class 1 acetylated histones. As most active chromatin and hyperacetylated histones are associated with the low salt-insoluble residual nuclear material containing the nuclear matrix, we investigated whether hyperacetylated histones are bound to the beta(A)- and epsilon-globin DNA in this fraction. In chromatin immunoprecipitation assays, we found that the beta(A)- and epsilon-globin coding regions are bound to hyperacetylated H3 and H4. Our observations are consistent with a model in which nuclear matrix-associated histone acetyltransferases and deacetylases mediate a dynamic attachment between active and competent chromatin and the nuclear matrix. PMID- 11435439 TI - Galectin-12, an Adipose-expressed Galectin-like Molecule Possessing Apoptosis inducing Activity. AB - Galectins constitute a family of proteins that bind to beta-galactoside residues and have diverse physiological functions. Here we report on the identification of a galectin-like molecule, galectin-12, in a human adipose tissue cDNA library. The protein contained two potential carbohydrate-recognition domains with the second carbohydrate-recognition domain being less conserved compared with other galectins. In vitro translated galectin-12 bound to a lactosyl-agarose column far less efficiently than galectin-8. Galectin-12 mRNA was predominantly expressed in adipose tissue of human and mouse and in differentiated 3T3-L1 adipocytes. Caloric restriction and treatment of obese animals with troglitazone increased galectin-12 mRNA levels and decreased the average size of the cells in adipose tissue. The induction of galectin-12 expression by the thiazolidinedione, troglitazone, was paralleled by an increase in the number of apoptotic cells in adipose tissue. Immunocytochemical analysis revealed that galectin-12 was localized in the nucleus of adipocytes, and transfection with galectin-12 cDNA induced apoptosis of COS-1 cells. These results suggest that galectin-12, an adipose-expressed galectin-like molecule, may participate in the apoptosis of adipocytes. PMID- 11435440 TI - Alternative use of a mini exon of the L1 gene affects L1 binding to neural ligands. AB - Neural cell adhesion molecule L1 is a cell surface glycoprotein required for the correct development of the nervous system. L1 exists as two isoforms encoded by mRNA species that either collectively incorporate or exclude exons 2 and 27. Neurons utilize only the full-length isoform, whereas Schwann cells, kidney cells, and blood lymphocytes only express the short form of L1. Still other cells, oligodendrocytes, regulate L1 isoform expression in a maturation-dependent manner. The RSLE motif encoded by exon 27 is known to have a role in clathrin mediated endocytosis of L1, but the function of the exon 2-encoded motif (YEGHHV) is unknown. Here we show that this motif is required for the optimal binding of L1 to several neural ligands and is likely to be important for nervous system development. Thus, alternative use of exon 2 is a mechanism for regulating ligand interactions with L1. PMID- 11435441 TI - Accelerated G(1) phase progression induced by the human T cell leukemia virus type I (HTLV-I) Tax oncoprotein. AB - Tax, the human T cell leukemia virus type I oncoprotein, plays a crucial role in viral transformation and the development of the virally associated disease adult T cell leukemia. Because oncogenesis involves alterations in cell growth, it is important to examine the effects of Tax on cell cycle progression. Using a synchronized cell system, we have found that Tax expression accelerates G(1) phase progression and S phase entry with concomitant DNA replication. This accelerated progression is accompanied by an earlier onset of cdk2 kinase activity. In contrast to the shortening of G(1) phase, the length of S phase is unaffected by Tax expression. As a result of a more rapid cell cycle progression, cells expressing Tax exhibit faster growth kinetics and display an altered cell cycle distribution. Additionally, the decreased time allowed for growth in the presence of Tax results in a decreased cell size. Tax-associated acceleration of cell cycle progression may play a role in the ability of this viral oncoprotein to mediate cellular transformation and promote the development of human T cell leukemia virus type I-associated diseases. PMID- 11435442 TI - RNA polymerase II elongator holoenzyme is composed of two discrete subcomplexes. AB - Elongator is a histone acetyltransferase complex that associates with the elongating form of RNA polymerase II. We purified Elongator to virtual homogeneity via a rapid three-step procedure based largely on affinity chromatography. The purified factor, holo-Elongator, is a labile six-subunit factor composed of two discrete subcomplexes: one comprised of the previously identified Elp1, Elp2, and Elp3 proteins and another comprised of three novel polypeptides, termed Elp4, Elp5, and Elp6. Disruption of the yeast genes encoding the new Elongator proteins confers phenotypes indistinguishable from those previously described for the other elp mutants, and concomitant disruption of genes encoding proteins in either subcomplex does not confer new phenotypes. Taken together, our results indicate that holo-Elongator is a functional entity in vitro as well as in vivo. Metazoan homologues of Elp1 and Elp3 have previously been reported. We cloned the human homologue of yeast ELP4 and show that this gene is ubiquitously expressed in human tissues. PMID- 11435443 TI - A role for calcium in stabilizing transport vesicle coats. AB - Calcium has been implicated in regulating vesicle fusion reactions, but its potential role in regulating other aspects of protein transport, such as vesicle assembly, is largely unexplored. We find that treating cells with the membrane permeable calcium chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl ester) (BAPTA-AM), leads to a dramatic redistribution of the vesicle coat protein, coatomer, in the cell. We have used the cell-free reconstitution of coat-protomer I (COPI) vesicle assembly to characterize the mechanisms of this redistribution. We find that the recovery of COPI-coated Golgi vesicles is inhibited by the addition of BAPTA to the cell-free vesicle budding assay. When coatomer-coated membranes are incubated in the presence of calcium chelators, the membranes "uncoat," indicating that calcium is necessary for maintaining the integrity of the coat. This uncoating is reversed by the addition of calcium. Interestingly, BAPTA, a calcium chelator with fast binding kinetics, is more potent at uncoating the coatomer-coated membrane than EGTA, suggesting that a calcium transient or a calcium gradient is important for stabilizing COPI vesicle coat. The primary target for the effects of calcium on coatomer recruitment is a step that occurs after ADP-ribosylation factor binding to the membrane. We suggest that a calcium gradient may serve to regulate the timing of vesicle uncoating. PMID- 11435444 TI - Dissociation of heparan sulfate and receptor binding domains of hepatocyte growth factor reveals that heparan sulfate-c-met interaction facilitates signaling. AB - Hepatocyte growth factor (HGF) is a secreted, heparan sulfate (HS) glycosaminoglycan-binding protein that stimulates mitogenesis, motogenesis, and morphogenesis in a wide array of cellular targets, including hepatocytes and other epithelial cells, melanocytes, endothelial cells, and hematopoietic cells. NK1 is an alternative HGF isoform that consists of the N-terminal (N) and first kringle (K1) domains of full-length HGF and stimulates all major HGF biological activities. Within NK1, the N domain retains the HS binding properties of full length HGF and mediates HS-stimulated ligand oligomerization but lacks significant mitogenic or motogenic activity. In contrast, K1 does not bind HS, but it stimulates receptor and mitogen-activated protein kinase activation, mitogenesis, and motogenesis, demonstrating that structurally distinct and dissociable domains of HGF are the primary mediators of HS binding and receptor activation. Despite the absence of HS-K1 binding, K1 mitogenic activity in HS negative cells is strictly dependent on added soluble heparin, whereas K1 stimulated motility is not. We also found that, like the receptors for fibroblast growth factors, the HGF receptor c-Met binds tightly to HS. These data suggest that HS can facilitate HGF signaling through interaction with c-Met that is independent of HGF-HS interaction and that the recruitment of specific intracellular effectors that mediate distinct HGF responses such as mitogenesis and motility is regulated by HS-c-Met interaction at the cell surface. PMID- 11435445 TI - Ligand-dependent interaction of estrogen receptor-alpha with members of the forkhead transcription factor family. AB - Estrogen acting through the estrogen receptor (ER) is able to regulate cell growth and differentiation of a variety of normal tissues and hormone-responsive tumors. Ligand-activated ER binds DNA and transactivates the promoters of estrogen target genes. In addition, ligand-activated ER can interact with other factors to alter the physiology and growth of cells. Using a yeast two-hybrid screen, we have identified an interaction between ER alpha and the proapoptotic forkhead transcription factor FKHR. The ER alpha-FKHR interaction depends on beta estradiol and is reduced significantly in the absence of hormone or the presence of Tamoxifen. A glutathione S-transferase pull-down assay was used to confirm the interaction and localized two interaction sites, one in the forkhead domain and a second in the carboxyl terminus. The FKHR interaction was specific to ER alpha and was not detected with other ligand-activated steroid receptors. The related family members, FKHRL1 and AFX, also bound to ER alpha in the presence of beta estradiol. FKHR augmented ER alpha transactivation through an estrogen response element. Conversely, ER alpha repressed FKHR-mediated transactivation through an insulin response sequence, and cell cycle arrest induced by FKHRL1 in MCF7 cells was abrogated by estradiol. These results suggest a novel mechanism of estrogen action that involves regulation of the proapoptotic forkhead transcription factors. PMID- 11435446 TI - Location of the receptor-interaction site on CheB, the methylesterase response regulator of bacterial chemotaxis. AB - Sensory adaptation in bacterial chemotaxis is mediated by covalent modification of chemoreceptors, specifically methylation and demethylation of glutamates catalyzed by methyltransferase CheR and methylesterase CheB. The methylesterase is a two-domain response regulator in which phosphorylation of the regulatory domain enhances activity of the catalytic domain. In Escherichia coli and Salmonella typhimurium, a crucial determinant of efficient methylation and demethylation is a specific pentapeptide sequence at the chemoreceptor carboxyl terminus, a position distant from sites of enzymatic action. Each enzyme binds pentapeptide, but the site of binding has been located only for CheR. Here we locate the pentapeptide-binding site on CheB by assessing catalytic activity and pentapeptide binding of CheB fragments, protection of CheB from proteolysis by pentapeptide, and interference with pentapeptide-CheB interaction by a CheB segment. The results place the binding site near the hinge between regulatory and catalytic domains, in a segment spanning the carboxyl-terminal end of the regulatory domain and the beginning of the linker that stretches to the catalytic domain. This location is quite different from the catalytic domain location of the pentapeptide-binding site on CheR and is likely to reflect the rather different ways in which pentapeptide binding enhances enzymatic action for the methyltransferase and the methylesterase. PMID- 11435447 TI - The serpins are an expanding superfamily of structurally similar but functionally diverse proteins. Evolution, mechanism of inhibition, novel functions, and a revised nomenclature. PMID- 11435448 TI - The T cell receptor for antigen: signaling and ligand discrimination. PMID- 11435449 TI - Golgi nucleotide sugar transport and leukocyte adhesion deficiency II. PMID- 11435450 TI - Cyclooxygenase-selective inhibition of prostanoid formation: transducing biochemical selectivity into clinical read-outs. PMID- 11435451 TI - Mixed messages: modulation of inflammation and immune responses by prostaglandins and thromboxanes. PMID- 11435452 TI - Genetic and pharmacological analysis of prostanoid receptor function. PMID- 11435453 TI - Immunomodulatory therapy of human type 1 diabetes: lessons from the mouse. PMID- 11435454 TI - The multiple contributions of thyroid hormone to heat production. PMID- 11435455 TI - Regulation of hypoxia-induced angiogenesis: a chaperone escorts VEGF to the dance. PMID- 11435456 TI - Expression of the oxygen-regulated protein ORP150 accelerates wound healing by modulating intracellular VEGF transport. AB - Expression of angiogenic factors such as VEGF under conditions of hypoxia or other kinds of cell stress contributes to neovascularization during wound healing. The inducible endoplasmic reticulum chaperone oxygen-regulated protein 150 (ORP150) is expressed in human wounds along with VEGF. Colocalization of these two molecules was observed in macrophages in the neovasculature, suggesting a role of ORP150 in the promotion of angiogenesis. Local administration of ORP150 sense adenovirus to wounds of diabetic mice, a treatment that efficiently targeted this gene product to the macrophages of wound beds, increased VEGF antigen in wounds and accelerated repair and neovascularization. In cultured human macrophages, inhibition of ORP150 expression caused retention of VEGF antigen within the endoplasmic reticulum (ER), while overexpression of ORP150 promoted the secretion of VEGF into hypoxic culture supernatants. Taken together, these data suggest an important role for ORP150 in the setting of impaired wound repair and identify a key, inducible chaperone-like molecule in the ER. This novel facet of the angiogenic response may be amenable to therapeutic manipulation. PMID- 11435457 TI - IFN-gamma and Fas/FasL are required for the antitumor and antiangiogenic effects of IL-12/pulse IL-2 therapy. AB - Systemic administration of IL-12 and intermittent doses of IL-2 induce complete regression of metastatic murine renal carcinoma. Here, we show that overt tumor regression induced by IL-12/pulse IL-2 is preceded by recruitment of CD8(+) T cells, vascular injury, disrupted tumor neovascularization, and apoptosis of both endothelial and tumor cells. The IL-12/IL-2 combination synergistically enhances cell surface FasL expression on CD8(+) T lymphocytes in vitro and induces Fas and FasL expression within tumors via an IFN-gamma-dependent mechanism in vivo. This therapy also inhibits tumor neovascularization and induces tumor regression by mechanisms that depend critically on endogenous IFN-gamma production and an intact Fas/FasL pathway. The ability of IL-12/pulse IL-2 to induce rapid destruction of tumor-associated endothelial cells and regression of established metastatic tumors is ablated in mice with a dysregulated Fas/FasL pathway. The common, critical role for endogenous IFN-gamma and the Fas/FasL pathway in early antiangiogenic effects and in antitumor responses suggests that early, cytokine driven innate immune mechanisms and CD8(+) T cell-mediated responses are interdependent. Definition of critical early molecular events engaged by IL-12/IL 2 may provide new perspective into optimal therapeutic engagement of a productive host-antitumor immune response. PMID- 11435458 TI - Reversal of established autoimmune diabetes by restoration of endogenous beta cell function. AB - In NOD (nonobese diabetic) mice, a model of autoimmune diabetes, various immunomodulatory interventions prevent progression to diabetes. However, after hyperglycemia is established, such interventions rarely alter the course of disease or allow sustained engraftment of islet transplants. A proteasome defect in lymphoid cells of NOD mice impairs the presentation of self antigens and increases the susceptibility of these cells to TNF-alpha-induced apoptosis. Here, we examine the hypothesis that induction of TNF-alpha expression combined with reeducation of newly emerging T cells with self antigens can interrupt autoimmunity. Hyperglycemic NOD mice were treated with CFA to induce TNF-alpha expression and were exposed to functional complexes of MHC class I molecules and antigenic peptides either by repeated injection of MHC class I matched splenocytes or by transplantation of islets from nonautoimmune donors. Hyperglycemia was controlled in animals injected with splenocytes by administration of insulin or, more effectively, by implantation of encapsulated islets. These interventions reversed the established beta cell-directed autoimmunity and restored endogenous pancreatic islet function to such an extent that normoglycemia was maintained in up to 75% of animals after discontinuation of treatment and removal of islet transplants. A therapy aimed at the selective elimination of autoreactive cells and the reeducation of T cells, when combined with control of glycemia, is thus able to effect an apparent cure of established type 1 diabetes in the NOD mouse. PMID- 11435459 TI - c-Jun N-terminal kinase is required for metalloproteinase expression and joint destruction in inflammatory arthritis. AB - Mitogen-activated protein kinase (MAPK) cascades are involved in inflammation and tissue destruction in rheumatoid arthritis (RA). In particular, c-Jun N-terminal kinase (JNK) is highly activated in RA fibroblast-like synoviocytes and synovium. However, defining the precise function of this kinase has been difficult because a selective JNK inhibitor has not been available. We now report the use of a novel selective JNK inhibitor and JNK knockout mice to determine the function of JNK in synoviocyte biology and inflammatory arthritis. The novel JNK inhibitor SP600125 (anthra[1,9-cd]pyrazol-6(2H)-one) completely blocked IL-1--induced accumulation of phospho-Jun and induction of c-Jun transcription in synoviocytes. Furthermore, AP-1 binding and collagenase mRNA accumulation were completely suppressed by SP600125. In contrast, complete inhibition of p38 had no effect, and ERK inhibition had only a modest effect. The essential role of JNK was confirmed in cultured synoviocytes from JNK1 knockout mice and JNK2 knockout mice, each of which had a partial defect in IL-1--induced AP-1 activation and collagenase-3 expression. Administration of SP600125 modestly decreased the rat paw swelling in rat adjuvant-induced arthritis. More striking was the near complete inhibition of radiographic damage that was associated with decreased AP 1 activity and collagenase-3 gene expression. Therefore, JNK is a critical MAPK pathway for IL-1--induced collagenase gene expression in synoviocytes and in joint arthritis, indicating that JNK is an important therapeutic target for RA. PMID- 11435460 TI - Systemic IFN-beta gene therapy results in long-term survival in mice with established colorectal liver metastases. AB - Most patients succumbing to colorectal cancer fail with liver-predominant metastases. To make a clinical impact in this disease, a systemic or whole-liver therapy may be required, whereas most cancer gene therapy approaches are limited in their ability to treat beyond local disease. As a preclinical model for cancer gene therapy, recombinant adenovirus containing the human IFN-beta (hIFN-beta) cDNA was delivered systemically in nude mouse xenograft models of human colorectal cancer liver metastases. The vector targeted hepatocytes that produced high levels of hIFN-beta in the liver, resulting in a profound apoptotic response in the tumors and significant tumor regression. hIFN-beta gene therapy not only resulted in improved survival and long-term cure in a micrometastatic model, but provided similar benefits in a clinically relevant gross disease model. A similar recombinant adenovirus containing the murine IFN-beta (mIFN-beta) cDNA also resulted in a therapeutic response and improved survival in syngeneic mouse models of colorectal cancer liver metastases. Depletion studies demonstrate a contribution of natural killer cells to this therapeutic response. The toxicity of an adenoviral vector expressing murine IFN-beta in a syngeneic model is also presented. These encouraging results warrant further investigation of the use of cancer gene therapy for targeting metastatic disease. PMID- 11435461 TI - Thyroid hormone--sympathetic interaction and adaptive thermogenesis are thyroid hormone receptor isoform--specific. AB - In newborns and small mammals, cold-induced adaptive (or nonshivering) thermogenesis is produced primarily in brown adipose tissue (BAT). Heat production is stimulated by the sympathetic nervous system, but it has an absolute requirement for thyroid hormone. We used the thyroid hormone receptor beta--selective (TR-beta--selective) ligand, GC-1, to determine by a pharmacological approach whether adaptive thermogenesis was TR isoform--specific. Hypothyroid mice were treated for 10 days with varying doses of T3 or GC-1. The level of uncoupling protein 1 (UCP1), the key thermogenic protein in BAT, was restored by either T3 or GC-1 treatment. However, whereas interscapular BAT in T3 treated mice showed a 3.0 degrees C elevation upon infusion of norepinephrine, indicating normal thermogenesis, the temperature did not increase (<0.5 degrees C) in GC-1--treated mice. When exposed to cold (4 degrees C), GC-1--treated mice also failed to maintain core body temperature and had reduced stimulation of BAT UCP1 mRNA, indicating impaired adrenergic responsiveness. Brown adipocytes isolated from hypothyroid mice replaced with T3, but not from those replaced with GC-1, had normal cAMP production in response to adrenergic stimulation in vitro. We conclude that two distinct thyroid-dependent pathways, stimulation of UCP1 and augmentation of adrenergic responsiveness, are mediated by different TR isoforms in the same tissue. PMID- 11435462 TI - Molecular basis of ocular abnormalities associated with proximal renal tubular acidosis. AB - Proximal renal tubular acidosis associated with ocular abnormalities such as band keratopathy, glaucoma, and cataracts is caused by mutations in the Na(+)-HCO(3)( ) cotransporter (NBC-1). However, the mechanism by which NBC-1 inactivation leads to such ocular abnormalities remains to be elucidated. By immunological analysis of human and rat eyes, we demonstrate that both kidney type (kNBC-1) and pancreatic type (pNBC-1) transporters are present in the corneal endothelium, trabecular meshwork, ciliary epithelium, and lens epithelium. In the human lens epithelial (HLE) cells, RT-PCR detected mRNAs of both kNBC-1 and pNBC-1. Although a Na(+)-HCO(3)-cotransport activity has not been detected in mammalian lens epithelia, cell pH (pH(i)) measurements revealed the presence of Cl(-) independent, electrogenic Na(+)-HCO(3)-cotransport activity in HLE cells. In addition, up to 80% of amiloride-insensitive pH(i) recovery from acid load in the presence of HCO(3)(-)/CO(2) was inhibited by adenovirus-mediated transfer of a specific hammerhead ribozyme against NBC-1, consistent with a major role of NBC-1 in overall HCO(3)-transport by the lens epithelium. These results indicate that the normal transport activity of NBC-1 is indispensable not only for the maintenance of corneal and lenticular transparency but also for the regulation of aqueous humor outflow. PMID- 11435463 TI - Familial CD8 deficiency due to a mutation in the CD8 alpha gene. AB - CD8 glycoproteins play an important role in both the maturation and function of MHC class I-restricted T lymphocytes. A 25-year-old man, from a consanguineous family, with recurrent bacterial infections and total absence of CD8(+) cells, was studied. Ab deficiencies and ZAP-70 and TAP defects were ruled out. A missense mutation (gly90-->ser) in both alleles of the immunoglobulin domain of the CD8 alpha gene was shown to correlate with the absence of CD8 expression found in the patient and two sisters. Conversely, high percentages of CD4(-)CD8( )TCR alpha beta(+) T cells were found in the three siblings. A novel autosomal recessive immunologic defect characterized by absence of CD8(+) cells is described. These findings may help to further understanding of the role of CD8 molecules in human immune response. PMID- 11435464 TI - Acetylcholine receptor delta subunit mutations underlie a fast-channel myasthenic syndrome and arthrogryposis multiplex congenita. AB - Limitation of movement during fetal development may lead to multiple joint contractures in the neonate, termed arthrogryposis multiplex congenita. Neuromuscular disorders are among the many different causes of reduced fetal movement. Many congenital myasthenic syndromes (CMSs) are due to mutations of the adult-specific epsilon subunit of the acetylcholine receptor (AChR), and, thus, functional deficits do not arise until late in gestation. However, an earlier effect on the fetus might be predicted with some defects of other AChR subunits. We studied a child who presented at birth with joint contractures and was subsequently found to have a CMS. Mutational screening revealed heteroallelic mutation within the AChR delta subunit gene, delta 756ins2 and delta E59K. Expression studies demonstrate that delta 756ins2 is a null mutation. By contrast, both fetal and adult AChR containing delta E59K have shorter than normal channel activations that predict fast decay of endplate currents. Thus, delta E59K causes dysfunction of fetal as well as the adult AChR and would explain the presence of joint contractures on the basis of reduced fetal movement. This is the first report of the association of AChR gene mutations with arthrogryposis multiplex congenita. It is probable that mutations that severely disrupt function of fetal AChR will underlie additional cases. PMID- 11435465 TI - Adenosine deaminase deficiency increases thymic apoptosis and causes defective T cell receptor signaling. AB - Adenosine deaminase (ADA) deficiency in humans results in a severe combined immunodeficiency (SCID). This immunodeficiency is associated with severe disturbances in purine metabolism that are thought to mediate lymphotoxicity. The recent generation of ADA-deficient (ADA(-/-)) mice has enabled the in vivo examination of mechanisms that may underlie the SCID resulting from ADA deficiency. We demonstrate severe depletion of T and B lymphocytes and defects in T and B cell development in ADA(-/-) mice. T cell apoptosis was abundant in thymi of ADA(-/-) mice, but no increase in apoptosis was detected in the spleen and lymph nodes of these animals, suggesting that the defect is specific to developing thymocytes. Studies of mature T cells recovered from spleens of ADA(-/ ) mice revealed that ADA deficiency is accompanied by TCR activation defects of T cells in vivo. Furthermore, ex vivo experiments on ADA(-/-) T cells demonstrated that elevated adenosine is responsible for this abnormal TCR signaling. These findings suggest that the metabolic disturbances seen in ADA(-/-) mice affect various signaling pathways that regulate thymocyte survival and function. Experiments with thymocytes ex vivo confirmed that ADA deficiency reduces tyrosine phosphorylation of TCR-associated signaling molecules and blocks TCR triggered calcium increases. PMID- 11435466 TI - Involvement of FAN in TNF-induced apoptosis. AB - TNF-alpha is a pleiotropic cytokine activating several signaling pathways initiated at distinct intracellular domains of the TNF receptors. Although the C terminal region is believed to be responsible for apoptosis induction, the functions of more membrane-proximal domains, including the domain that couples to neutral sphingomyelinase activation, are not yet fully elucidated. The roles of this region and of the associated adapter protein FAN (factor associated with neutral SMase activation) in the cytotoxic response to TNF have been investigated. We have now shown that stable expression in human fibroblasts of a dominant negative form of FAN abrogates TNF-induced ceramide generation from sphingomyelin hydrolysis and reduces caspase processing, thus markedly inhibiting TNF-triggered apoptosis. However, the cytotoxic responses to daunorubicin and exogenous ceramide remain unaltered, as do the TNF-induced p42/p44 MAPK activation and CD54 expression. Fibroblasts from FAN-knockout mice also proved to be resistant to TNF toxicity. These findings highlight the previously unrecognized role of the adapter protein FAN in signaling cell death induction by TNF. PMID- 11435467 TI - Glucose toxicity and the development of diabetes in mice with muscle-specific inactivation of GLUT4. AB - Using cre/loxP gene targeting, transgenic mice with muscle-specific inactivation of the GLUT4 gene (muscle GLUT4 KO) were generated and shown to develop a diabetes phenotype. To determine the mechanism, we examined insulin-stimulated glucose uptake and metabolism during hyperinsulinemic-euglycemic clamp in control and muscle GLUT4 KO mice before and after development of diabetes. Insulin stimulated whole body glucose uptake was decreased by 55% in muscle GLUT4 KO mice, an effect that could be attributed to a 92% decrease in insulin-stimulated muscle glucose uptake. Surprisingly, insulin's ability to stimulate adipose tissue glucose uptake and suppress hepatic glucose production was significantly impaired in muscle GLUT4 KO mice. To address whether these latter changes were caused by glucose toxicity, we treated muscle GLUT4 KO mice with phloridzin to prevent hyperglycemia and found that insulin-stimulated whole body and skeletal muscle glucose uptake were decreased substantially, whereas insulin-stimulated glucose uptake in adipose tissue and suppression of hepatic glucose production were normal after phloridzin treatment. In conclusion, these findings demonstrate that a primary defect in muscle glucose transport can lead to secondary defects in insulin action in adipose tissue and liver due to glucose toxicity. These secondary defects contribute to insulin resistance and to the development of diabetes. PMID- 11435468 TI - Discrete cleavage motifs of constitutive and immunoproteasomes revealed by quantitative analysis of cleavage products. AB - Proteasomes are the main proteases responsible for cytosolic protein degradation and the production of major histocompatibility complex class I ligands. Incorporation of the interferon gamma--inducible subunits low molecular weight protein (LMP)-2, LMP-7, and multicatalytic endopeptidase complex--like (MECL)-1 leads to the formation of immunoproteasomes which have been associated with more efficient class I antigen processing. Although differences in cleavage specificities of constitutive and immunoproteasomes have been observed frequently, cleavage motifs have not been described previously. We now report that cells expressing immunoproteasomes display a different peptide repertoire changing the overall cytotoxic T cell--specificity as indicated by the observation that LMP-7(-/-) mice react against cells of LMP-7 wild-type mice. Moreover, using the 436 amino acid protein enolase-1 as an unmodified model substrate in combination with a quantitative approach, we analyzed a large collection of peptides generated by either set of proteasomes. Inspection of the amino acids flanking proteasomal cleavage sites allowed the description of two different cleavage motifs. These motifs finally explain recent findings describing differential processing of epitopes by constitutive and immunoproteasomes and are important to the understanding of peripheral T cell tolerization/activation as well as for effective vaccine development. PMID- 11435469 TI - Anuria, omphalocele, and perinatal lethality in mice lacking the CD34-related protein podocalyxin. AB - Podocalyxin is a CD34-related sialomucin that is expressed at high levels by podocytes, and also by mesothelial cells, vascular endothelia, platelets, and hematopoietic stem cells. To elucidate the function of podocalyxin, we generated podocalyxin-deficient (podxl(-/)-) mice by homologous recombination. Null mice exhibit profound defects in kidney development and die within 24 hours of birth with anuric renal failure. Although podocytes are present in the glomeruli of the podxl(-/)- mice, they fail to form foot processes and slit diaphragms and instead exhibit cell--cell junctional complexes (tight and adherens junctions). The corresponding reduction in permeable, glomerular filtration surface area presumably leads to the observed block in urine production. In addition, podxl( /)- mice frequently display herniation of the gut (omphalocele), suggesting that podocalyxin may be required for retraction of the gut from the umbilical cord during development. Hematopoietic and vascular endothelial cells develop normally in the podocalyxin-deficient mice, possibly through functional compensation by other sialomucins (such as CD34). Our results provide the first example of an essential role for a sialomucin in development and suggest that defects in podocalyxin could play a role in podocyte dysfunction in renal failure and omphalocele in humans. PMID- 11435470 TI - Murine cytomegalovirus is regulated by a discrete subset of natural killer cells reactive with monoclonal antibody to Ly49H. AB - Antiviral roles of natural killer (NK) cell subsets were examined in C57BL/6 mice infected with murine cytomegalovirus (MCMV) and other viruses, including lymphocytic choriomeningitis virus (LCMV), vaccinia virus (VV), and mouse hepatitis virus (MHV). Each virus vigorously induced an NK cell infiltrate into the peritoneal cavity and liver, causing some redistributions of NK cell subsets defined by monoclonal antibody (mAb) directed against Ly49A, C/I, D, and G2. Striking results were seen with a mAb (1F8) reactive with the positively signaling molecule Ly49H, present in MCMV-resistant C57BL/6 mice. mAb 1F8 also stains Ly49 C and I, but exclusion of those reactivities with mAb 5E6, which recognizes Ly49 C and I, indicated that Ly49H(+) cells infiltrated the peritoneal cavity and liver and were particularly effective at synthesizing interferon gamma. Depletion of 1F8(+) but not 5E6(+) cells in vivo by mAb injections enhanced MCMV titers by 20-1,000-fold in the spleen and approximately fivefold in the liver. Titers of LCMV or VV were not enhanced. These anti-MCMV effects were attributed to prototypical NK1.1(+)CD3(-) NK cells and not to NK1.1(+)CD3(+) "NK/T" cells. This is the first evidence that control of a virus infection in vivo is mediated by a distinct NK cell subset. PMID- 11435471 TI - A coordinated change in chemokine responsiveness guides plasma cell movements. AB - Antibody-secreting plasma cells are nonrecirculatory and lodge in splenic red pulp, lymph node medullary cords, and bone marrow. The factors that regulate plasma cell localization are poorly defined. Here we demonstrate that, compared with their B cell precursors, plasma cells exhibit increased chemotactic sensitivity to the CXCR4 ligand CXCL12. At the same time, they downregulate CXCR5 and CCR7 and have reduced responsiveness to the B and T zone chemokines CXCL13, CCL19, and CCL21. We demonstrate that CXCL12 is expressed within splenic red pulp and lymph node medullary cords as well as in bone marrow. In chimeric mice reconstituted with CXCR4-deficient fetal liver cells, plasma cells are mislocalized in the spleen, found in elevated numbers in blood, and fail to accumulate normally in the bone marrow. Our findings indicate that as B cells differentiate into plasma cells they undergo a coordinated change in chemokine responsiveness that regulates their movements in secondary lymphoid organs and promotes lodgment within the bone marrow. PMID- 11435472 TI - Hyperactivation of p21(ras) and the hematopoietic-specific Rho GTPase, Rac2, cooperate to alter the proliferation of neurofibromin-deficient mast cells in vivo and in vitro. AB - Mutations in the NF1 tumor suppressor gene cause neurofibromatosis type I (NF1), a disease characterized by the formation of cutaneous neurofibromas infiltrated with a high density of degranulating mast cells. A hallmark of cell lines generated from NF1 patients or Nf1-deficient mice is their propensity to hyperproliferate. Neurofibromin, the protein encoded by NF1, negatively regulates p21(ras) activity by accelerating the conversion of Ras-GTP to Ras-GDP. However, identification of alterations in specific p21(ras) effector pathways that control proliferation in NF1-deficient cells is incomplete and critical for understanding disease pathogenesis. Recent studies have suggested that the proliferative effects of p21(ras) may depend on signaling outputs from the small Rho GTPases, Rac and Rho, but the physiologic importance of these interactions in an animal disease model has not been established. Using a genetic intercross between Nf1(+/)- and Rac2(-)(/)- mice, we now provide genetic evidence to support a biochemical model where hyperactivation of the extracellular signal-regulated kinase (ERK) via the hematopoietic-specific Rho GTPase, Rac2, directly contributes to the hyperproliferation of Nf1-deficient mast cells in vitro and in vivo. Further, we demonstrate that Rac2 functions as mediator of cross-talk between phosphoinositide 3-kinase (PI-3K) and the classical p21(ras)-Raf-Mek-ERK pathway to confer a distinct proliferative advantage to Nf1(+/)- mast cells. Thus, these studies identify Rac2 as a novel mediator of cross-talk between PI-3K and the p21(ras)-ERK pathway which functions to alter the cellular phenotype of a cell lineage involved in the pathologic complications of a common genetic disease. PMID- 11435473 TI - Brain mast cells act as an immune gate to the hypothalamic-pituitary-adrenal axis in dogs. AB - Mast cells perform a significant role in the host defense against parasitic and some bacterial infections. Here we show that in the dog, degranulation of brain mast cells evokes hypothalamic-pituitary-adrenal responses via histamine release. A large number of mast cells were found in a circumscribed ventral region of the hypothalamus, including the pars tuberalis and median eminence. When these intracranial mast cells were passively sensitized with immunoglobulin E via either the intracerebroventricular or intravenous route, there was a marked increase in the adrenal cortisol secretion elicited by a subsequent antigenic challenge (whether this was delivered via the central or peripheral route). Comp.48/80, a mast cell secretagogue, also increased cortisol secretion when administered intracerebroventricularly. Pretreatment (intracerebroventricularly) with anti-corticotropin--releasing factor antibodies or a histamine H(1) blocker, but not an H(2) blocker, attenuated the evoked increases in cortisol. These data show that in the dog, degranulation of brain mast cells evokes hypothalamic pituitary-adrenal responses via centrally released histamine and corticotrophin releasing factor. On the basis of these data, we suggest that intracranial mast cells may act as an allergen sensor, and that the activated adrenocortical response may represent a life-saving host defense reaction to a type I allergy. PMID- 11435474 TI - Molecular genetic analysis of an endotoxin nonresponder mutant cell line: a point mutation in a conserved region of MD-2 abolishes endotoxin-induced signaling. AB - Somatic cell mutagenesis is a powerful tool for characterizing receptor systems. We reported previously two complementation groups of mutant cell lines derived from CD14-transfected Chinese hamster ovary--K1 fibroblasts defective in responses to bacterial endotoxin. Both classes of mutants expressed a normal gene product for Toll-like receptor (TLR)4, and fully responded to stimulation by tumor necrosis factor (TNF)-alpha or interleukin (IL)-1 beta. We identified the lesion in one of the complementation groups in the gene for MD-2, a putative TLR4 coreceptor. The nonresponder phenotype of this mutant was reversed by transfection with MD-2. Cloning of MD-2 from the nonresponder cell line revealed a point mutation in a highly conserved region resulting in a C95Y amino acid exchange. Both forms of MD-2 colocalized with TLR4 on the cell surface after transfection, but only the wild-type cDNA reverted the lipopolysaccharide (LPS) nonresponder phenotype. Furthermore, soluble MD-2, but not soluble MD-2(C95Y), functioned to enable LPS responses in cells that expressed TLR4. Thus, MD-2 is a required component of the LPS signaling complex and can function as a soluble receptor for cells that do not otherwise express it. We hypothesize that MD-2 conformationally affects the extracellular domain of TLR4, perhaps resulting in a change in affinity for LPS or functioning as a portion of the true ligand for TLR4. PMID- 11435475 TI - Tumor necrosis factor receptor-associated factor (TRAF)2 represses the T helper cell type 2 response through interaction with NFAT-interacting protein (NIP45). AB - Recently we have identified a novel protein NIP45 (nuclear factor of activated T cells [NFAT]-interacting protein) which substantially augments interleukin (IL)-4 gene transcription. The provision of NIP45 together with NFAT and the T helper cell type 2 (Th2)-specific transcription factor c-Maf to cells normally refractory to IL-4 production, such as B cells or Th1 clones, results in substantial IL-4 secretion to levels that approximate those produced by primary Th2 cells. In studies designed to further our understanding of NIP45 activity, we have uncovered a novel facet of IL-4 gene regulation. We present evidence that members of the tumor necrosis factor receptor-associated factor (TRAF) family of proteins, generally known to function as adapter proteins that transduce signals from the tumor necrosis factor receptor superfamily, contribute to the repression of IL-4 gene transcription and that this effect is mediated through their interaction with NIP45. PMID- 11435476 TI - Separation of Notch1 promoted lineage commitment and expansion/transformation in developing T cells. AB - Notch1 signaling is required for T cell development. We have previously demonstrated that expression of a dominant active Notch1 (ICN1) transgene in hematopoietic stem cells (HSCs) leads to thymic-independent development of CD4(+)CD8(+) double-positive (DP) T cells in the bone marrow (BM). To understand the function of Notch1 in early stages of T cell development, we assessed the ability of ICN1 to induce extrathymic T lineage commitment in BM progenitors from mice that varied in their capacity to form a functional pre-T cell receptor (TCR). Whereas mice repopulated with ICN1 transduced HSCs from either recombinase deficient (Rag-2(-/)-) or Src homology 2 domain--containing leukocyte protein of 76 kD (SLP-76)(-/)- mice failed to develop DP BM cells, recipients of ICN1 transduced Rag-2(-/)- progenitors contained two novel BM cell populations indicative of pre-DP T cell development. These novel BM populations are characterized by their expression of CD3 epsilon and pre-T alpha mRNA and the surface proteins CD44 and CD25. In contrast, complementation of Rag-2(-/)- mice with a TCR beta transgene restored ICN1-induced DP development in the BM within 3 wk after BM transfer (BMT). At later time points, this population selectively and consistently gave rise to T cell leukemia. These findings demonstrate that Notch signaling directs T lineage commitment from multipotent progenitor cells; however, both expansion and leukemic transformation of this population are dependent on T cell-specific signals associated with development of DP thymocytes. PMID- 11435477 TI - The epithelial cellular adhesion molecule (Ep-CAM) is a ligand for the leukocyte associated immunoglobulin-like receptor (LAIR). AB - Human leukocyte-associated immunoglobulin-like receptor (LAIR)-1 is expressed on many cells of the immune system and is predicted to mediate inhibitory functions based on the presence of immunoreceptor tyrosine-based inhibitory motifs (ITIMs) in its cytoplasmic domain. Although the role of LAIR-1 in the regulation of immune responses in vivo is unknown, LAIR-1 cross-linking by monoclonal antibody inhibits various immune cell functions in vitro. Here, we identify the colon carcinoma-associated epithelial cellular adhesion molecule (Ep-CAM) as a ligand for LAIR-1 and LAIR-2, a related soluble LAIR-1 family member. Ep-CAM interacts with the LAIR molecules through its first epidermal growth factor domain; Ep-CAM- specific antibodies can abrogate the binding. Intraepithelial T lymphocytes express LAIR-1 and thus may interact with Ep-CAM present on human intestinal epithelium. We propose that LAIR-1--Ep-CAM interaction may contribute to mucosal tolerance and that LAIR-2 possibly modulates this function. PMID- 11435478 TI - The diverse roles of mast cells. PMID- 11435479 TI - The search for new vaccines against tuberculosis. AB - The failure of the BCG vaccine for tuberculosis in large, controlled clinical trials, coupled with the gradual consensus that it is mostly ineffective in preventing adult pulmonary disease in endemic areas, has led to a concerted effort to develop a new generation of vaccines. This work is ongoing in a variety of areas, including DNA vaccines, subunit vaccines, recombinant vaccines, and auxotrophic vaccines. Several such candidates are giving promising results in mouse and guinea pig, aerosol-challenge infection models and should move to clinical trials in the near future. PMID- 11435480 TI - Transmembrane proteases in focus: diversity and redundancy? AB - Recent advances have led to the identification and characterization of an array of transmembrane proteases that mediate the proteolysis of various substrates (including bioactive peptides, components of the extracellular matrix, and integral proteins) and cell-cell or cell-matrix adhesion. The membrane proteases known to participate in these processes currently include the ectopeptidases, the membrane-type matrix metalloproteases (MT-MMPs), the ADAM (a disintegrin and metalloprotease) family, the meprins, and the secretases, and this list may be expected to grow. The roles that these molecules play within neoplastic and inflammatory sites are being investigated actively. The capacity of these ectoenzymes to transmit intracellular-transduction signals through the plasma membrane has to be considered. An appreciation of their functional redundancy is emerging. PMID- 11435481 TI - Unlocking the secrets of cytotoxic granule proteins. AB - Cytotoxic lymphocytes largely comprise CD8(+) cytotoxic T cells and natural killer cells and form the major defense of higher organisms against virus infected and transformed cells. A key function of cytotoxic lymphocytes is to detect and eliminate potentially harmful cells by inducing them to undergo apoptosis. This is achieved through two principal pathways, both of which require direct but transient contact between the killer cell and its target. The first, involving ligation of TNF receptor-like molecules such as Fas/CD95 by their cognate ligands, results in mobilization of conventional, programmed cell-death pathways centered on activation of pro-apoptotic caspases. This review concentrates on the second pathway, in which the toxic contents of secretory vesicles of the cytotoxic lymphocyte are secreted toward the target cell, and some toxins penetrate into the target cell cytoplasm and nucleus. In addition to invoking a powerful stimulus to caspase activation, this "granule-exocytosis mechanism" provides a variety of additional strategies for overcoming inhibitors of the caspase cascade that may be elaborated by viruses. The key molecular players in this process are the pore-forming protein perforin and a family of granule-bound serine proteases or granzymes. The molecular functions of perforin and granzymes are under intense investigation in many laboratories including our own, and recent advances will be discussed. In addition, this review discusses the evidence pointing to the importance of perforin and granzyme function in pathophysiological situations as diverse as infection with intracellular pathogens, graft versus host disease, susceptibility to transplantable and spontaneous malignancies, lymphoid homeostasis, and the tendency to auto-immune diseases. PMID- 11435482 TI - Long-term-impaired expression of nuclear factor-kappa B and I kappa B alpha in peripheral blood mononuclear cells of trauma patients. AB - Nuclear factor (NF)-kappa B expression and dimer characteristics were studied in peripheral blood mononuclear cells (PBMCs) of major-trauma patients and healthy controls. Analysis of PBMCs on days 1, 3, 5, and 10 after trauma revealed that expression of both p65p50 heterodimers and p50p50 homodimers was significantly reduced compared with that in controls. In vitro lipopolysaccharide (LPS) stimulation of PBMCs induced NF-kappa B translocation. However, throughout the survey, p65p50 activation remained significantly lower in trauma patients than in controls. After LPS stimulation in vitro, the p65p50/p50p50 ratio was significantly lower in PBMCs from trauma patients than from healthy controls. The ex vivo expression of I kappa B alpha was higher in PBMCs of controls than of trauma patients. LPS did not induce I kappa B expression in PBMCs from trauma patients, but strong induction was obtained with staphylococci, suggesting that this defect is not universal and depends on the nature of the activating signal. Although no direct correlation was found between levels of interleukin-10 or transforming growth factor-beta and NF-kappa B, these immunosuppressive cytokines were significantly elevated in trauma patients by 10 days after admission. The long-term low-basal and LPS-induced nuclear translocation of NF-kappa B recalled long-term immunoparalysis observed in patients with severe inflammatory stress such as trauma. PMID- 11435483 TI - Effects of macrophage-CSF on pulmonary-macrophage repopulation after bone marrow transplantation. AB - Pulmonary infections are important causes of morbidity and mortality in immunosuppressed patients after transplantation. After experimental irradiation and syngeneic bone marrow transplantation in mice, macrophages show reduced repopulation in the lung compared with that in other tissues. Macrophages are major microbicidal immune effector cells in host pulmonary defense. Therefore, we examined the role of locally applied cytokines for macrophage repopulation in the lung. An accelerated repopulation of macrophages in the lung was observed after intranasal application of macrophage-colony stimulating factor (M-CSF), but this effect was not enhanced by a combination of M-CSF with interleukin (IL)-3. Local proliferation contributed to this effect. Macrophages in the lung tissue of M-CSF treated mice displayed greater secretion of IL-6, whereas M-CSF treatment did not enhance the gene expression of other macrophage-specific chemokines. The role of M-CSF treatment was determined in pulmonary murine cytomegalovirus infection using an irradiation/reconstitution model. The M-CSF treatment had no effect on virus load in the lung tissue. However, phosphate-buffered saline-treated mice seemed to develop stronger inflammation after viral infection than M-CSF-treated mice. We conclude that local M-CSF treatment modulates cellular inflammation in the lung during immunosuppression. PMID- 11435484 TI - Immunocytochemical localization of peptidylarginine deiminase in human eosinophils and neutrophils. AB - Peptidylarginine deiminase, registered as PAD V in the DDBJ/GenBank/EMBL data banks, is expressed in HL-60 cells differentiated into granulocytes or monocytes. We analyzed PAD activities in density-fractionated human peripheral blood cell fractions. PAD activity with similar substrate specificity to that of PAD V was found in the eosinophil and neutrophil fractions, which showed single bands comigrating with authentic PAD V on immunoblotting with an anti-PAD V antibody. Both the biochemical and immunoblotting analyses showed marked enrichment of PAD V in the eosinophil fraction. Its immunoreactivity appeared to localize in eosinophilic granules at high density and in myeloperoxidase-negative cytoplasmic granules of neutrophils at low density, as determined by confocal laser-scanning microscopy. Possible roles of PAD V in myeloid differentiation and granulocyte function are discussed. In addition, we present evidence for the presence of PAD(s) that are antigenically different from PAD V in monocytes and lymphocytes. PMID- 11435485 TI - Retardation of early-onset PMA-induced apoptosis in mouse neutrophils deficient in myeloperoxidase. AB - Neutrophil apoptosis is a mechanism involved in the resolution of inflammation. To explore the role of hypochlorous acid (HOCl) produced by neutrophils while they are undergoing apoptosis, we compared the rates of apoptosis in neutrophils isolated from normal mice and from myeloperoxidase (MPO)-deficient mice, which are unable to generate HOCl. Apoptosis in MPO-deficient neutrophils stimulated by phorbol myristate acetate (PMA) was significantly slower than in normal neutrophils during 3 h of incubation. Exposure of normal neutrophils to H(2)O(2) together with PMA resulted in a dramatic acceleration of apoptosis, and almost all of the cells revealed apoptotic morphology at 1 h. This acceleration was inhibited by cytochrome c, a superoxide scavenger. Conversely, in MPO-deficient neutrophils activated with PMA and H(2)O(2), little acceleration was observed before 1 h, although it gradually increased thereafter. This retardation was almost completely reversed when MPO or HOCl was exogenously added. These results suggest that coexistence of HOCl and superoxide accelerates the early onset of neutrophil apoptosis. PMID- 11435486 TI - Dimeric S100A8 in human neutrophils is diminished after phagocytosis. AB - S100A8 is a major cytoplasmic protein of neutrophils and monocytes/macrophages and has been associated with myeloid cell differentiation and activation. Little is known about its functions or mechanisms of release from neutrophils. We have developed a monoclonal antibody to murine S100A8, which cross-reacts with human S100A8. This antibody, which recognizes the homodimeric form of the protein, detects its expression specifically in human neutrophils and is reactive in formalin-fixed, paraffin-embedded tissues. Using this antibody as well as a commercially available antibody to human S100A8, we show that phagocytic activation of neutrophils, in vivo in acute appendicitis and in vitro following phagocytosis of opsonized zymosan, is characterized by loss of cytoplasmic immunoreactivity for S100A8. In vitro, phagocytosis is associated with rapid diminution of immunostaining without loss of viability. Loss of immunoreactivity for S100A8 may serve as a marker of localized neutrophil activation in tissues. PMID- 11435487 TI - Serotonin protects NK cells against oxidatively induced functional inhibition and apoptosis. AB - High concentrations of the neurotransmitter serotonin can be found in inflamed and ischemic peripheral tissues, but the role of serotonin in immunoregulation is largely unknown. Here we report that serotonin protected human natural-killer (NK) cells from oxidatively induced inhibition inflicted by autologous monocytes in vitro. Serotonin protected NK cells from monocyte-mediated apoptosis and suppression of cytotoxicity and maintained the activation of NK cells induced by interleukin-2 despite the presence of inhibitory monocytes. A detailed analysis of these protective effects revealed that serotonin scavenged reactive oxygen species (ROS) derived from the H(2)O(2)-myeloperoxidase (-MPO) system. Serotonin shared this scavenger activity with its precursor, 5-hydroxytryptophan (5-HTP); however, serotonin was >10-fold more potent than 5-HTP in protecting NK cells against functional inhibition and apoptosis. We propose that serotonin, by scavenging peroxidase-derived ROS, may serve to protect NK cells from oxidative damage at inflammatory sites. PMID- 11435488 TI - Regulated expression of galectin-1 during B-cell activation and implications for T-cell apoptosis. AB - Galectin-1 (GAL-1), a highly conserved beta-galactoside-binding protein, has shown immunomodulatory properties. In this study, we investigated the regulation of GAL-1 expression within the B-cell compartment using Trypanosoma cruzi infection as a natural model of in vivo B-cell activation. GAL-1 was found to be expressed on activated B cells from T. cruzi-infected mice, mainly localized at the cytosolic compartment. Expression of this protein was found to be modulated according to the activation state of the cells, revealing a significant increase in stimulated B cells that received signals via cross-linking of the B-cell receptor and CD40. It was found that GAL-1 was secreted by B cells to the extracellular milieu upon activation. Finally, purified GAL-1 produced by activated B cells induced apoptosis of T cells but not B cells and also influenced interferon-gamma cytokine production. Hence, the present study describes a potential mechanism by which B cells can regulate T-cell function and survival. PMID- 11435489 TI - Induction of interleukin-8 in human neutrophils after MHC class II cross-linking with superantigens. AB - Neutrophils have been shown to express major histocompatibility complex class II (MHC II) after stimulation. However, reports concerning the functional effect of MCH II expression are still lacking. In our hands, granulocyte-monocyte colony stimulating factor (GM-CSF) alone and in combination with interferon (IFN)-gamma, but not IFN-gamma or interleukin (IL)-3, induced a significant level of expression of human leukocyte antigen DR on neutrophils. The addition of staphylococcal enterotoxin E to neutrophils resulted in a significant increase in IL-8 production only after prestimulation with GM-CSF alone or in combination with IFN-gamma but had no effect on neutrophils preincubated with IFN-gamma alone or IL-3. Staphylococcal enterotoxin A, another bivalent superantigen, also stimulated production of IL-8 by preincubated polymorphonuclear neutrophils, whereas staphylococcal enterotoxin A mutants that are not able to cross-link MHC II molecules failed to induce IL-8 production. Taken together, our results clearly demonstrate that after induction of MHC II, neutrophils are able to respond to MHC II-specific stimulation. These findings support the ideas that the induced MHC II complex is completely functional and that neutrophils may be able to present antigens. PMID- 11435491 TI - Stimulation of human neutrophils and monocytes by staphylococcal phenol-soluble modulin. AB - Modulins represent microbial products that stimulate cytokine production in host cells. The modulins responsible for gram-positive sepsis remain poorly understood. Staphylococci release a factor (or factors) that activates nuclear factor-kappa B and stimulates cytokine production in cells of macrophage lineage. This factor, termed phenol-soluble modulin (PSM), has been recently isolated from culture supernatant of Staphylococcus epidermidis. We examined the effects of PSM on proinflammatory properties of human neutrophils and monocytes in vitro. PSM activated the respiratory (oxidative) burst in neutrophils and primed neutrophils for enhanced respiratory burst activity in response to formyl-methionyl-leucyl phenylalanine. PSM also stimulated neutrophil degranulation as reflected by increased surface expression of CD11b and CD18, which was accompanied by rapid shedding of L-selectin. Spontaneous apoptosis of both neutrophils and monocytes was inhibited by PSM. Furthermore, PSM also functioned as a chemoattractant factor for both neutrophils and monocytes. Thus, the proinflammatory properties of PSM resemble those of both lipopolysaccharide and bacterial chemotactic peptides. These findings suggest that PSM may play a role in the pathogenesis and systemic manifestations of sepsis caused by staphylococci. PMID- 11435490 TI - Nitric oxide induces murine thymocyte apoptosis by oxidative injury and a p53 dependent mechanism. AB - Previously, we showed that NO induces thymocyte apoptosis via a caspase-1 dependent mechanism [(1) ]. In the present study, we investigated the role of heme oxygenase, catalase, bax, and p53 in this process. The NO donor, S-nitroso-N acetyl penicillamine (SNAP), induced DNA fragmentation in thymocytes in a time- and concentration-dependent way. SNAP (100 microM) induced 50--60% apoptosis; higher doses did not increase the rate of apoptosis significantly. SNAP decreased catalase and heme iron (Fe) levels without affecting superoxide dismutase, glutathione, or total Fe stores in thymocytes. SNAP significantly increased the expression of heme oxygenase 1 (HSP-32), p53, and bax but not bcl-2. Treatment with the heme oxygenase inhibitor, tin protoporphyrin IX inhibited SNAP-induced thymocyte apoptosis. Furthermore, thymocytes from p53 null mice were resistant to NO-induced apoptosis. Our data suggest that NO may induce its cytotoxic effects on thymocytes by modulating heme oxygenase and catalase activity as well as up regulating pro-apoptotic proteins p53 and bax. PMID- 11435492 TI - Immunostimulatory CpG-modified plasmid DNA enhances IL-12, TNF-alpha, and NO production by bovine macrophages. AB - The immunogenicity of DNA vaccines is partially attributable to the adjuvant properties of bacterial plasmid DNA (pDNA) for B lymphocytes and professional antigen-presenting cells. In mice, modification of immunostimulatory sequences (ISSs), including CpG motifs, in pDNA vectors or oligodeoxynucleotides can increase or decrease their adjuvant properties. ISSs that stimulate optimal responses reportedly differ for murine and human leukocytes. We have previously characterized the mitogenic properties of oligodeoxynucleotides containing one AACGTT motif for bovine B lymphocytes. We now define cytokine responses by macrophages stimulated with pDNA engineered to contain an ISS comprising two AACGTT motifs. Macrophages activated with CpG-modified pDNA secreted significantly more interleukin-12, tumor necrosis factor-alpha, and nitric oxide than macrophages stimulated with unmodified pDNA or modified pDNA that contained nucleotides scrambled to remove CpG motifs. Engineered CpG-pDNA or CpG oligodeoxynucleotides should be useful as vaccines or adjuvants to promote the enhanced type 1 responses important for protection against intracellular pathogens. PMID- 11435493 TI - Chemokine receptors in human basophils: inducible expression of functional CXCR4. AB - We examined the expression profile of chemokine receptors in human basophils and their regulation by cytokines. Basophils expressed transcripts of CC chemokine receptors (CCR)1, CCR2, CCR3, and CCR5 and CXC chemokine receptors (CXCR)1, CXCR2, and CXCR4. In contrast to the other receptors, surface-CXCR4 expression was not detected in fresh- and whole-blood basophils, but it became apparent gradually during incubation. Among 16 chemokines tested, eotaxin induced the most potent basophil migration. SDF-1 also induced a strong, migratory response comparable with that induced by eotaxin in 24-h, cultured basophils, but it failed to induce degranulation. IL-3 abrogated CXCR4 expression completely, and it only down-regulated CCR2 and CCR3 expression slightly. IL-5, GM-CSF, and IL-4 also down-regulated CXCR4 expression. Thus, expression of CXCR4 was the most strongly affected by cytokines, and this may represent an alternative mechanism for control of cell-specific, biological responses to SDF-1. PMID- 11435494 TI - Potentiation of interferon-gamma-stimulated nitric oxide production by retinoic acid in RAW 264.7 cells. AB - Nitric oxide (NO) production is essential for normal immunity. We have examined the capacity of retinoic acid (RA), a pleiotropic hormone necessary for normal immunity, to modulate NO production in RAW 264.7 cells. NO production induced by suboptimal concentrations of interferon-gamma (IFN-gamma) was significantly greater in cells cultured in low-retinoid medium and treated with all-trans-RA (10(-10) - 10(-6) M, P <0.05), as well as with 9-cis-RA and several retinoids selective for the RA receptor subfamily of nuclear retinoid receptors. Similar results were obtained with lipopolysaccharide and monophosphoryl lipid A as stimuli. The RA-potentiated production of NO was positively correlated with inducible NO synthase (iNOS) protein (r =0.94, P <0.002), although the expression of iNOS mRNA was not altered. We hypothesize that modulation of the macrophage response to suboptimal immune stimuli by physiological concentrations of RA, as observed in these studies, may be important in establishing an optimal balance between T helper (Th) 1- and Th2-mediated immunity. PMID- 11435495 TI - Mechanism of extracellular release of human neutrophil calprotectin complex. AB - Calprotectin is an abundant cytosolic protein complex of human neutrophils with in vitro extracellular antimicrobial activity. Studies suggest that calprotectin may be actively secreted from intact HL-60 cells and that it can be translocated to polymorphonuclear neutrophil (PMN) cell membranes. To examine whether calprotectin is secreted extracellularly, we incubated soluble and particulate stimuli, including live and heat-inactivated Candida albicans, with whole blood and measured calprotectin levels in the plasma. We compared the release of calprotectin to that of lactoferrin, a protein known to be secreted by PMNs. Extracellular lactoferrin was detected after incubation with any of the particulate stimuli. In contrast, a significant increase in extracellular calprotectin was found only after incubation with live C. albicans. Specifically, the increase in extracellular calprotectin correlated directly with a proportional decrease in PMN viability. Our results indicate that human PMN calprotectin is not secreted extracellularly except as a result of cell disruption or death. PMID- 11435496 TI - Surfactant protein D-coated Klebsiella pneumoniae stimulates cytokine production in mononuclear phagocytes. AB - Encapsulated Klebsiella pneumoniae strains K21a, K10, and K50, all of which contain dimannose sequences in their capsular polysaccharides that are recognized by the mannose receptor of macrophages, stimulated interleukin secretion and cytokine mRNA expression by human monocyte-derived macrophages. By contrast, the corresponding unencapsulated phase variants and the K2 strain, which lack the dimannose sequence, did not. Coating of unencapsulated phase variants of Klebsiella strains with surfactant protein (SP)-D resulted in marked stimulation of cytokine mRNA accumulation. The induction of cytokine mRNA via the mannose receptor occurred only in monocyte-derived macrophages, whereas that caused by SP D-coated Klebsiella strains occurred in both macrophages and peripheral-blood monocytes. The results suggested that innate immunity against pulmonary pathogens might be mediated by SP-D, which acts as an opsonin to enhance the interaction of macrophages with unencapsulated phase variants originating from the upper respiratory tract, and by macrophage mannose receptors, which recognize encapsulated variants expressing capsular dimannose residues. PMID- 11435497 TI - Signaling pathways initiated in macrophages after engagement of type A scavenger receptors. AB - Scavenger receptors are macrophage cell surface molecules associated with endocytic uptake of lipoproteins and binding of microbial ligands. Macrophage class A scavenger receptors (SR-As) interact with ligands to induce cellular signaling leading to gene transcription and cytokine release. We used inhibitors of early and late signaling to block SR-A-mediated polyinosinic-polycytidilic acid (poly I:C) and lipoteichoic acid (LTA) activation of RAW 264.7 macrophages. Effects of multiple inhibitors on tumor necrosis factor (TNF)-alpha release were monitored to determine requirements for inflammatory cytokine production. Cycloheximide, monodansylcadaverine, and cytochalasin B all blocked TNF-alpha release from macrophages stimulated with LTA or poly I:C, whereas monensin only nominally reduced TNF-alpha production. Selected inhibitors of downstream signaling events reduced SR-A-dependent TNF-alpha release by >95% after stimulation with either ligand, whereas others were ineffective. The PKC inhibitor H7 reduced LTA-dependent secretion of TNF-alpha by 94% but inhibited poly I:C-dependent TNF-alpha production only by 50%. Priming of RAW 264.7 cells with interferon-gamma potentiated the response to poly I:C but did not alter inhibitor effects. These results demonstrated that for both ligands tested here, early events of receptor internalization are requisite for cellular activation. The response pattern suggests that tyrosine phosphorylation and activation of the MAP kinase pathway are key components of SR-A-mediated signal transduction cascades. PMID- 11435498 TI - Role of extracellular signal-regulated protein kinase cascade in macrophage killing of Candida albicans. AB - The pathogenic yeast Candida albicans and its derived molecules stimulate a wide range of macrophage secretory functions and may adapt to escape being killed by this phagocyte. In this study, phagocytosis of C. albicans and of the nonpathogenic yeast Saccharomyces cerevisiae was shown to be associated with phosphorylation of the mitogen-activated protein kinase (MAPK)/extracellularly regulated kinase (ERK) pathway in the absence of significant activation of either p38MAPK or stress-activated protein kinase/c-Jun N-terminal kinase. However, although 80% of endocytosed C. albicans survived after 1 h, 80% of S. cerevisiae cells were killed. Considerable quantitative differences were observed between the two species in the sequential phosphorylation of MAPK/ERK kinase (MEK), extracellularly regulated kinase-1, and 90-kDa-ribosomal S6 kinases. A lower level of activation of the pathway by C. albicans was associated with a species specific overexpression of the MEK phosphatase MAPK phosphatase (MKP)-1. Killing of both C. albicans and S. cerevisiae could be reduced using PD98059, which mimics MKP-1 and inhibits MEK phosphorylation, suggesting that specific MKP-1 activation by C. albicans could contribute to its ability to escape the yeast lytic potential of macrophages. PMID- 11435499 TI - Synthetic peptide MMK-1 is a highly specific chemotactic agonist for leukocyte FPRL1. AB - Human phagocytic leukocytes express the seven-transmembrane G-protein-coupled receptors formyl peptide receptor (FPR) and FPR-like 1 (FPRL1). MMK-1, a synthetic peptide derived from a random peptide library, is reported to induce calcium mobilization specifically in human FPRL1 gene-transfected cells. However, its actions on human phagocytic leukocytes remain poorly defined. We found that MMK-1 is a potent chemotactic and calcium-mobilizing agonist for human monocytes, neutrophils, and FPRL1-transfected human embryonic kidney (HEK) 293 cells but is inactive in cells transfected with FPR. MMK-1 also activated HEK 293 cells transfected with FPR2, a mouse counterpart of human FPRL1. Furthermore, MMK-1 increased pertussis toxin-sensitive production of inflammatory cytokines in human monocytes. MMK-1 signaling in human phagocytes was completely desensitized by a well-defined FPRL1 agonist, suggesting that FPRL1 is likely a receptor that mediates the action of MMK-1 in primary cells. Since MMK-1 is one of the most potent FPRL1-specific agonists identified so far, it can serve as a modulator of the host defense and a useful agent for further studying the signaling and function of FPRL1. PMID- 11435500 TI - Dairy food consumption, blood pressure and stroke. AB - Recent clinical and biochemical evidence supporting the hypothesis that consumption of dairy products may be associated with reduced blood pressure and risk of stroke is reviewed. The two prospective studies of dairy food consumption and stroke incidence both indicate that a higher intake of dairy foods reduces risk. It is difficult to associate any one mineral in dairy products to reduction in blood pressure or stroke incidence because an appropriate metabolic balance of all three is important and because of the strong correlations among Ca, Mg and K intakes when dairy products are consumed. In fact, the evidence reviewed indicates that although K apparently has the greatest effect, all three minerals potentially contribute to blood pressure and stroke reduction, i.e., a dietary balance of all three is recommended. Milk and food products such as yogurt made from milk, which retain substantial amounts of K, Ca and Mg, are important dietary sources of all three of these minerals. In addition, milk is a low Na food, which, as seen in Dietary Approaches to Stop Hypertension (DASH) II, provides further benefit in blood pressure reduction. New studies have associated dairy food consumption with other potential mechanisms affecting stroke, mainly reduction of platelet aggregation and insulin resistance. Further research is required to explore the relationship of dairy food consumption and stroke. PMID- 11435501 TI - The challenges of promoting optimal infant growth. PMID- 11435502 TI - African American dietary patterns at the beginning of the 20th century. AB - Early field studies in human nutrition documented the eating habits of African Americans living in a variety of circumstances. We compare the results of these investigations. Our analysis shows systematic differences along a continuum reaching from remote, rural communities in the South toward increasingly metropolitan locations. On the latter end of the continuum, we find diets richer in protein, composed of a wider variety of foods and containing fewer of what we now call "soul foods." Greater market involvement and access to low cost alternatives to more traditional foods help explain these developments. PMID- 11435503 TI - Dietary spermidine and spermine participate in the maturation of galactosyltransferase activity and glycoprotein galactosylation in rat small intestine. AB - This study considered the role of dietary polyamines in the maturation of intestinal glycoprotein galactosylation during postnatal development. In the rat small intestine, O-glycan: beta-1,3-galactosyltransferase and N-glycan: beta-1,4 galactosyltransferase are, respectively, involved in the glycan chain biosynthesis of mucins and of glycoproteins in the brush border membranes. Their activities increase significantly at weaning, in parallel with a rise in the intestinal content of spermidine and spermine (as determined by high performance liquid chromatography) and in proportion to the polyamine increase in food intake. The oral ingestion of spermidine or spermine (at 0.4 micromol/g body) by immature suckling rats for 4 d reproduced the levels of spermine and spermidine in their intestines at the time of weaning and induced precocious and significant rises in O-glycan: and N-glycan: galactosyltransferase activities to those normally found after weaning. In parallel, more galactose residues (detected in the complex oligosaccharide chains of glycoproteins by specific lectins after electrophoresis and transfer to nitrocellulose membranes) were observed in the brush border membranes of spermidine- and spermine-treated rats. In contrast, the ingestion of putrescine or ornithine had no effect. Diets with different levels of polyamines (milks and commercial diet), when given at weaning, induced variable evolutions of the galactosylation process, partly in relation to the amounts of polyamines ingested. These results indicate that spermidine and spermine are maturation factors that can reproduce, in immature rats, the same increase in intestinal glycoprotein galactosylation that is normally observed during weaning. They also suggest that the maturation of glycoprotein galactosylation may be a multifactorial event in which spermidine and spermine are both involved. PMID- 11435504 TI - Dietary fats affect rat plasma lipoprotein secondary structure as assessed by Fourier transform infrared spectroscopy. AB - This investigation was undertaken to determine by Fourier transform infrared spectroscopy the effects of diets enriched with fish, sunflower or olive oils on the secondary structure of plasma HDL and LDL from rats, as well as the effects on lipid unsaturation and acyl chain lengths. Controls were fed a commercial diet. In HDL, random coil conformation was relatively high in rats fed the fish diet, probably due to the irregular geometry of polyunsaturated fatty acids interacting with apoproteins. Parallel structural behaviors were observed for rats fed control and olive oil diets. The lowest lipid unsaturation level was found in HDL of rats fed olive oil, and acyl chain lengths were slightly increased by the three fats. Rats fed olive oil had the lowest percentage of LDL beta-sheets and these were more abundant in rats fed the fish oil diet. The least lipid unsaturation in LDL was in rats fed the olive oil diet. No significant differences in acyl chain lengths were observed. Certain protein conformational changes and/or apoprotein composition differences due to dietary fat may affect the binding between lipoproteins and their receptors in cells. PMID- 11435505 TI - Carnitine alters binding of aflatoxin to DNA and proteins in rat hepatocytes and cell-free systems. AB - The objective of this study was to determine effects of L-carnitine on aflatoxin B(1) (AFB(1))-DNA adduct formation in isolated rat hepatocytes, its dose response, specificity and mode of action. All experiments were conducted in either freshly isolated rat hepatocytes or cell-free systems. There was negative linear correlation between the dosage of carnitine and formation of [(3)H]AFB(1) DNA adducts in the hepatocytes; however, the partitioning of AFB(1) into cellular compartments was not affected by carnitine. The attenuating effect of carnitine on AFB(1)-DNA adduct formation was also present in a cell-free system, but there was lack of specificity because acetylcarnitine and gamma-aminobutyric acid (GABA) were equally effective. Carnitine appears to interfere with bioactivation of AFB(1) and binding of AFB(1)-epoxide to DNA. On the contrary, carnitine enhanced the binding of AFB(1) and its epoxide to microsomal proteins, plasma proteins and bovine serum albumin. These results indicate that carnitine diverts AFB(1)-epoxide away from DNA by promoting binding to proteins. We conclude that modulation of AFB(1) binding to proteins and DNA by carnitine alters the carcinogenic and hepatotoxic potential of AFB(1) and poses concerns about the human AFB(1)-exposure data based on the AFB(1)-albumin adduct concentrations as a biomarker. PMID- 11435506 TI - Biotin regulates the genetic expression of holocarboxylase synthetase and mitochondrial carboxylases in rats. AB - Biotin is the cofactor of carboxylases [pyruvate (PC), propionyl-CoA (PCC), 3 methyl crotonyl-CoA and acetyl-CoA], to which it is covalently bound by the action of holocarboxylase synthetase (HCS). We have studied whether biotin also regulates their expression, as it does other, nonrelated enzymes (e.g., glucokinase, phosphoenol pyruvate carboxykinase, guanylate cyclase). For this purpose, HCS, PC and PCC mRNAs were studied in biotin-deficient rat liver, kidney, muscle and brain of biotin-deficient rats. PC- and PCC-specific activities and protein masses were also measured. The 24-h time course of HCS mRNA in deficient rats was examined after biotin supplementation. HCS mRNA was significantly reduced during vitamin deficiency. It increased in deficient rats after biotin was injected, reaching control levels 24 h after administration. These changes seem to be the first known instance in mammals of an effect of a water-soluble vitamin on a mRNA functionally related to it. In contrast, the decreased activities of the carboxylases were associated with reductions in the amounts of their enzyme proteins except in brain. However, their mRNA levels were not affected. There are no reports on these types of vitamin affecting the mRNA or protein levels of their apoenzymes or their products. This work provides evidence for biotin being a modulator of the genetic expression of the enzymes involved in its function as a cofactor. As such, it may be a useful model for probing a similar role for other water-soluble vitamins. PMID- 11435507 TI - Retinoid compounds activate and induce hepatic glycine N-methyltransferase in rats. AB - Glycine N-methyltransferase (GNMT) functions to regulate S-adenosylmethionine (SAM) levels and the ratio of SAM/S-adenosylhomocysteine (SAH). SAM is a universal methyl group donor and the up-regulation of GNMT may lead to wastage of methyl groups required for transmethylation reactions. Previously, we demonstrated that dietary treatment of rats with 13-cis-retinoic acid (CRA) decreased the hepatic concentration of SAM and the SAH ratio. Here, we examined the ability of CRA, as well as all-trans-retinoic acid (ATRA), to regulate hepatic GNMT as a potential basis for our earlier observations. Rats were fed either a control (10% casein + 0.3% L-methionine) diet or a control diet supplemented with L-methionine (10 g/kg diet). Rats from each group were orally given ATRA, CRA (both at 30 micromol/kg body), or vehicle daily for 7 d. For control rats, administration of both CRA and ATRA elevated the hepatic GNMT activity 49% and 34%, respectively, compared with the control group. Similar results were exhibited by rats fed the methionine-supplemented diet. Moreover, the retinoid-induced elevations in enzyme activity were reflected in the abundance of GNMT protein. To our knowledge, this is the first report of a nutritional compound that induces GNMT activity at the transcriptional and/or translational level. PMID- 11435508 TI - Dietary supplementation with gamma-linolenic acid or fish oil decreases T lymphocyte proliferation in healthy older humans. AB - Animal and human studies have shown that greatly increasing the amounts of flax seed oil [rich in the (n-3) polyunsaturated fatty acid (PUFA) alpha-linolenic acid (ALNA)] or fish oil [FO; rich in the long chain (n-3) PUFA eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)] in the diet can decrease mitogen stimulated lymphocyte proliferation. The objective of this study was to determine the effect of dietary supplementation with moderate levels of ALNA, gamma linolenic acid (GLA), arachidonic acid (ARA), DHA or FO on the proliferation of mitogen-stimulated human peripheral blood mononuclear cells (PBMC) and on the production of cytokines by those cells. The study was randomized, placebo controlled, double-blinded and parallel. Healthy subjects ages 55-75 y consumed nine capsules/d for 12 wk; the capsules contained placebo oil (an 80:20 mix of palm and sunflower seed oils) or blends of placebo oil with oils rich in ALNA, GLA, ARA or DHA or FO. Subjects in these groups consumed 2 g of ALNA or 770 mg of GLA or 680 mg of ARA or 720 mg of DHA or 1 g of EPA plus DHA (720 mg of EPA + 280 mg of DHA) daily from the capsules. Total fat intake from the capsules was 4 g/d. The fatty acid composition of PBMC phospholipids was significantly changed in the GLA, ARA, DHA and FO groups. Lymphocyte proliferation was not significantly affected by the placebo, ALNA, ARA or DHA treatments. GLA and FO caused a significant decrease (up to 65%) in lymphocyte proliferation. This decrease was partly reversed by 4 wk after stopping the supplementation. None of the treatments affected the production of interleukin-2 or interferon-gamma by PBMC and none of the treatments affected the number or proportion of T or B lymphocytes, helper or cytotoxic T lymphocytes or memory helper T lymphocytes in the circulation. We conclude that a moderate level GLA or EPA but not of other (n 6) or (n-3) PUFA can decrease lymphocyte proliferation but not production of interleukin-2 or interferon-gamma. PMID- 11435509 TI - Kinetics of folate turnover in pregnant women (second trimester) and nonpregnant controls during folic acid supplementation: stable-isotopic labeling of plasma folate, urinary folate and folate catabolites shows subtle effects of pregnancy on turnover of folate pools. AB - To investigate the effects of pregnancy on folate metabolism, we conducted an 84 d study in second-trimester (gestational wk 14-25) pregnant women (n = 6) and nonpregnant controls (n = 6) with stable-isotopic tracer methods. All subjects were fed a diet containing approximately 272 nmol/d (120 microg/d) folate from food, along with supplemental folic acid that contained 15% [3',5'-(2)H(2)] folic acid ([(2)H(2)]folic acid) during d 1--41 and that was unlabeled during d 42--84 to yield a constant total folate intake of 1.02 or 1.93 micromol/d (450 or 850 microg/d). Isotopic enrichment of plasma folate, urinary folate and the urinary folate catabolites para-aminobenzoylglutamate (pABG) and para acetamidobenzoylglutamate (ApABG) was determined at intervals throughout the study. The labeling of pABG and ApABG reflected that of tissue folate pools from which the catabolites originate. After the intake of labeled folic acid was terminated on d 41, labeling of urinary folate exhibited a biphasic exponential decline with distinct fast and slow components. In contrast, during d 42--84, the enrichment of urinary pABG and ApABG exhibited primarily monophasic exponential decline, and plasma folate underwent little decline of labeling during this period. Pregnant women and controls did not differ in estimates of body folate pool size and most aspects of the excretion of labeled urinary folate and catabolites, rates of decline of excretion, and areas under the curves for folate and catabolite excretion. Pregnant women, however, tended to have a slower rate of decline of pABG than ApABG and higher enrichment at d 42 of ApABG and pABG. These data support and extend our previous findings indicating that pregnancy (gestational wk 14--26) causes subtle changes in folate metabolism but does not elicit substantial increases in the rate or extent of folate turnover at these moderately high folate intakes. PMID- 11435510 TI - Quercetin glucuronides but not glucosides are present in human plasma after consumption of quercetin-3-glucoside or quercetin-4'-glucoside. AB - The nature of quercetin conjugates present in blood after consumption of quercetin glucosides is still unclear. In this study, we analyzed plasma of volunteers that had consumed 325 micromol of either quercetin-3-glucoside or quercetin-4'-glucoside as an oral solution. Quercetin metabolites were extracted with acetonitrile/phosphoric acid and these extracts were analyzed using a high performance liquid chromatography with Coularray detection that distinguishes between the glucuronidated and the glucosylated forms of quercetin. No intact quercetin glucosides and only trace amounts of aglycone were found in human plasma, irrespective of the glucoside ingested. This was confirmed by spiking the plasma with glucoside standards. The major components in plasma had the same retention time as quercetin glucuronide standards. These plasma components disappeared after treatment of the plasma with bovine liver beta-glucuronidase, under reformation of quercetin, and showed the same oxidation pattern as the glucuronides. These results suggest that after consumption of quercetin glucosides, quercetin glucuronides are major metabolites in plasma. PMID- 11435511 TI - Dietary plant sterols alter the serum plant sterol concentration but not the cholesterol precursor sterol concentrations in young children (the STRIP Study). Special Turku Coronary Risk Factor Intervention Project. AB - Plant sterol supplementation reduces serum cholesterol concentration but may increase serum plant sterol concentrations, especially in children. We determined whether natural dietary plant sterols derived mainly from vegetable oil or margarine in early childhood affect serum concentrations of plant sterols (campesterol and sitosterol) and cholesterol precursor sterols (Delta-8 cholestenol, desmosterol, and lathosterol), reflecting endogenous cholesterol synthesis. We measured the serum sterol concentrations using gas liquid chromatography in 20 healthy 13-mo-old intervention children in a randomized, prospective study designed to decrease exposure of the children to known environmental atherosclerosis risk factors and in 20 control children. The diet of the intervention children was rich in plant sterols due to replacement of milk fat with vegetable fat, whereas the diet of the control children contained only small amounts of plant sterols. The intervention children consumed twice as much plant sterols as the control children (P < 0.001). Their serum concentrations of campesterol and sitosterol were 75% and 44% higher, respectively, than those in the control children (P < 0.001 for both), but serum cholesterol precursor sterol concentrations did not differ between the two groups. We conclude that doubling dietary plant sterol intake almost doubles serum plant sterol concentrations in 13-mo-old children, but has no effect on endogenous cholesterol synthesis. Relative intestinal absorption of natural plant sterols from the diet in early childhood is similar to that in adults. PMID- 11435512 TI - Food supplementation with encouragement to feed it to infants from 4 to 12 months of age has a small impact on weight gain. AB - It is unclear whether a substantial decline in malnutrition among infants in developing countries can be achieved by increasing food availability and nutrition counseling without concurrent morbidity-reducing interventions. The study was designed to determine whether provision of generous amounts of a micronutrient-fortified food supplement supported by counseling or nutritional counseling alone would significantly improve physical growth between 4 and 12 mo of age. In a controlled trial, 418 infants 4 mo of age were individually randomized to one of the four groups and followed until 12 mo of age. The first group received a milk-based cereal and nutritional counseling; the second group monthly nutritional counseling alone. To control for the effect of twice-weekly home visits for morbidity ascertainment, similar visits were made in one of the control groups (visitation group); the fourth group received no intervention. The median energy intake from nonbreast milk sources was higher in the food supplementation group than in the visitation group by 1212 kJ at 26 wk (P < 0.001), 1739 kJ at 38 wk (P < 0.001) and 2257 kJ at 52 wk (P < 0.001). The food supplementation infants gained 250 g (95% confidence interval: 20--480 g) more weight than did the visitation group. The difference in the mean increment in length during the study was 0.4 cm (95% confidence interval: -0.1--0.9 cm). The nutritional counseling group had higher energy intakes ranging from 280 to 752 kJ at different ages (P < 0.05 at all ages) but no significant benefit on weight and length increments. Methods to enhance the impact of these interventions need to be identified. PMID- 11435513 TI - Fruit and vegetable consumption is lower and saturated fat intake is higher among Canadians reporting smoking. AB - Understanding differences in dietary patterns by smoking status is important for nutritionists and health educators involved in helping individuals to make healthy dietary and lifestyle choices. Although smokers have a poor quality diet compared with nonsmokers, no study has examined nutritional adequacy and variability in the nutrient intake of smokers. The aim of this study was to compare dietary habits of smokers with nonsmokers in terms of nutrient intake, food groups contributing to nutrient intake, nutritional adequacy and day-to-day variation in nutrient intake. Noninstitutionalized adults aged 18--65 y (n = 1543) who participated in the Food Habits of Canadians Survey (1997--1998) were studied. Subjects, selected from across Canada using a multistage, random sampling strategy, completed an in-home 24-h dietary recall. Repeat interviews were conducted in a subsample to estimate variability in nutrient intake. Smokers had higher intakes of total and saturated fat, and lower intakes of folate, vitamin C and fiber than nonsmokers. There were no significant differences in calcium, zinc and vitamin A intakes or day-to-day variation in nutrient intake by smoking status. Smokers consumed significantly fewer fruits and vegetables than nonsmokers, leading to lower intakes of folate and vitamin C. In conclusion, smokers have a less healthy diet than nonsmokers, placing them at higher risk for chronic disease as a result of both dietary and smoking habits. Diet may act as a confounder in smoking-disease relationships. PMID- 11435514 TI - Dietary protein restriction and fat supplementation diminish the acidogenic effect of exercise during repeated sprints in horses. AB - A restricted protein diet supplemented with amino acids and fat may reduce the acidogenic effects of exercise. Twelve Arabian horses were assigned to a 2 x 2 factorial experiment: two fat levels: 0 or 10 g/100 g added corn oil and two crude protein levels: 7.5 g/100 g (supplemented with 0.5% L-lysine and 0.3% L threonine) or 14.5 g/100 g. The experiment began with a 4-wk diet accommodation period followed by a standard exercise test consisting of six 1-minute sprints at 7 m/s. Horses were interval trained for 11 wk followed by another exercise test with sprints at 10 m/s. Blood samples were taken at rest and during the exercise tests. Plasma was analyzed for PCO(2), PO(2), Na(+), K(+), Cl(-), lactate, pH and total protein. Bicarbonate, strong ion difference and total weak acids were calculated. Data were analyzed using repeated-measures analysis of variance. Venous pH was higher in the low protein group during the first test (P = 0.0056) and strong ion difference became higher (P = 0.022) during sprints in the low protein group. During the second test, venous pH and bicarbonate were higher for the low protein high fat group (P = 0.022 and P = 0.043, respectively) and strong ion difference became higher (P = 0.038) at the end of exercise in the low protein groups. These results show that restriction of dietary protein diminishes the acidogenic effect of exercise, especially in combination with fat adaptation. PMID- 11435515 TI - Whole-body nitrogen and splanchnic amino acid metabolism differ in rats fed mixed diets containing casein or its corresponding amino acid mixture. AB - Whole-body and splanchnic metabolism of dietary amino acids derived from casein (CAS) or the corresponding crystalline L-amino acid mixture (AA) were compared. Male adult rats were adapted for 9 d to two isoenergetic, isonitrogenous diets (15 g/100 g protein, 5 g/100 g fat) containing either CAS or AA. On d 10, the rats were fed a single mixed meal (3 g dry mass) containing either intrinsically (13)C-labeled goat casein or the amino acid mixture containing [U-(13)C(6)] leucine and [alpha-(15)N] lysine. Rats were killed before and 1, 3, 5 and 7 h after meal ingestion and samples of plasma, stomach wall and contents, small intestine and liver were collected. (13)C and (15)N enrichments of free and protein-bound amino acids in plasma and tissues were analyzed by gas chromatography-combustion isotope ratio mass spectrometry. Urinary nitrogen excretion was higher (P < 0.05) and weight gain lower (P < 0.05) in rats given the AA diet, indicating a lower whole-body net protein synthesis. Free (13)C leucine from the AA diet appeared in the intestinal mucosa free pool more rapidly (P < 0.05) than the CAS-(13)C-leucine, probably due to the faster transit through the stomach of the AA group. However, the incorporation of dietary leucine into plasma and liver proteins was higher in the CAS group 7 h after the meal (P < 0.05), whereas lysine incorporation into liver protein was higher in the AA group (P < 0.05). We conclude that whole-body protein homeostasis is better supported by dietary casein-bound than crystalline free amino acids, and that protein-bound leucine, but not lysine, is used more efficiently for liver protein synthesis than dietary free leucine. PMID- 11435516 TI - Acetic acid feeding enhances glycogen repletion in liver and skeletal muscle of rats. AB - To investigate the efficacy of the ingestion of vinegar in aiding recovery from fatigue, we examined the effect of dietary acetic acid, the main component of vinegar, on glycogen repletion in rats. Rats were allowed access to a commercial diet twice daily for 6 d. After 15 h of food deprivation, they were either killed immediately or given 2 g of a diet containing 0 (control), 0.1, 0.2 or 0.4 g acetic acid/100 g diet for 2 h. The 0.2 g acetic acid group had significantly greater liver and gastrocnemius muscle glycogen concentration than the control group (P < 0.05). The concentrations of citrate in this group in both the liver and skeletal muscles were >1.3-fold greater than in the control group (P > 0.1). In liver, the concentration of xylulose-5-phosphate in the control group was significantly higher than in the 0.2 and 0.4 g acetic acid groups (P < 0.01). In gastrocnemius muscle, the concentration of glucose-6-phosphate in the control group was significantly lower and the ratio of fructose-1,6-bisphosphate/fructose 6-phosphate was significantly higher than in the 0.2 g acetic acid group (P < 0.05). This ratio in the soleus muscle of the acetic acid fed groups was <0.8 fold that of the control group (P > 0.1). In liver, acetic acid may activate gluconeogenesis and inactivate glycolysis through inactivation of fructose-2,6 bisphosphate synthesis due to suppression of xylulose-5-phosphate accumulation. In skeletal muscle, acetic acid may inhibit glycolysis by suppression of phosphofructokinase-1 activity. We conclude that a diet containing acetic acid may enhance glycogen repletion in liver and skeletal muscle. PMID- 11435517 TI - The ratio of insoluble to soluble fiber components in soybean hulls affects ileal and total-tract nutrient digestibilities and fecal characteristics of dogs. AB - An experiment was conducted to evaluate the effects of soybean hulls (SH) containing varying ratios of insoluble:soluble fiber (I:S) on nutrient digestibilities and fecal characteristics of dogs. Ileally cannulated dogs (n = 6) were fed seven diets in a 6 x 7 Youden square arrangement of treatments. The seven diets included five SH-containing diets with I:S of 1.9, 2.7, 3.2, 5.2 or 7.2 and two diets containing either beet pulp (BP) or no supplemental fiber (control). Ileal digestibilities of DM, OM, CP, total dietary fiber (TDF), fat and gross energy (GE) were lower (P < 0.05) for dogs fed diets containing supplemental fiber compared with dogs fed the control diet. Fiber inclusion had a modest negative effect (P < 0.05) on total-tract DM, OM, fat and GE digestibilities compared with the control diet. Ileal digestibilities of DM and OM by dogs fed the SH treatments responded quadratically (P < 0.05) to I:S, with digestibility coefficients decreasing as the I:S approached 3.2. Highest ileal digestibilities were observed for diets with an I:S of 1.9 and 7.2. Similarly, a quadratic response (P < 0.05) was observed for digestibility of total amino acids at the ileum. Fecal outputs were lower (P < 0.001) when dogs consumed the control diet vs. fiber-containing diets. Among the SH-containing diets, there was a linear increase in fecal output as I:S increased (P = 0.031). The I:S in the diet affects DM and OM digestibilities at the ileum and affects fecal output, indicating that optimization of this ratio is desirable. PMID- 11435518 TI - Butyrate impairs energy metabolism in isolated perfused liver of fed rats. AB - This study was designed to test the effects of short-chain fatty acids (SCFA) with an even number of carbon atoms on hepatic energy metabolism. The effect of the SCFA was evaluated by measuring liver ATP content and oxygen consumption. The ATP content was evaluated using (31)P nuclear magnetic resonance in isolated liver from fed rats. In addition, respiratory activity (VO(2)) was assessed using Clark electrodes. The livers were perfused with acetate, butyrate or a medium chain length fatty acid, octanoate, at a concentration of 0.05--5.0 mmol/L. The addition of each substrate enhanced the rate of the net ATP consumption (V(i)), establishing a new ATP steady state that required a perfusion time of > or = 20 min, dependent on the chain length and concentration of the fatty acid (FA). The initial V(i) was unchanged for acetate and the ATP level stabilized at 58% of the initial level. Both butyrate and octanoate induced a dose-dependent increase in V(i). This may reflect an ATP-consuming process for the intracellular pH regulation observed during the acidosis associated with the beta-oxidation pathway. At the new steady state, the ATP concentration was approximately 45% of the initial level for both FA. VO(2) was both rapidly and reversibly increased, and the change was a function of butyrate or octanoate concentration and of the chain length. K(m) values were similar for butyrate and octanoate. Because all of the effects were similar for butyrate and octanoate, in contrast to acetate, we suggest that the impairment of the energy metabolism by butyrate resulted from an increase in the FADH(2)/NADH ratio due to beta-oxidation. In conclusion, the difference in the hepatic oxidation pathways of two products of intestinal fermentation (acetate and butyrate) explains their different actions on energy metabolism. PMID- 11435519 TI - The in vivo fate of hydroxytyrosol and tyrosol, antioxidant phenolic constituents of olive oil, after intravenous and oral dosing of labeled compounds to rats. AB - In vitro studies have shown phenolics in olive oil to be strong radical scavengers. The absorption and elimination of two radiolabeled phenolic constituents of olive oil, hydroxytyrosol and tyrosol were studied in vivo using rats. Compounds were administered intravenously (in saline) and orally (in oil- and water-based solutions). For both compounds, the intravenously and orally administered oil-based dosings resulted in significantly greater elimination of the phenolics in urine within 24 h than the oral, aqueous dosing method. There was no significant difference in the amount of phenolic compounds eliminated in urine between the intravenous dosing method and the oral oil-based dosing method for either tyrosol or hydroxytyrosol. Oral bioavailability estimates of hydroxytyrosol when administered in an olive oil solution and when dosed as an aqueous solution were 99% and 75%, respectively. Oral bioavailability estimates of tyrosol, when orally administered in an olive oil solution and when dosed as an aqueous solution were 98% and 71%, respectively. This is the first study that has used a radiolabeled compound to study the in vivo biological fates of hydroxytyrosol and tyrosol. PMID- 11435520 TI - Ascorbic acid synthesis in fetal and neonatal pigs and in pregnant and postpartum sows. AB - The ontogeny of ascorbic acid synthesis and its concentration in fetal pigs from mid- to late gestation, and the effect of birth order and premature or normal delivery ages were evaluated. In Experiment 1, fetal pigs were collected from three sows at 60, 80, 100, 107 and 111 d of development. Liver L-gulono-gamma lactone oxidase (GLO) activity and ascorbic acid concentration were measured. High liver GLO activity in fetal liver occurred at 60 d but declined as pregnancy advanced (P < 0.01), whereas ascorbic acid concentration increased (P < 0.01). Experiment 2 evaluated ascorbic acid synthesis and concentration in neonates born early (1st and 2nd) or late (7th and 8th) in the birthing sequence, or when born 2 d prematurely vs. the normal delivery age. Pigs born early in the birthing sequence (P < 0.01) and those born at the natural delivery age (P < 0.05) had higher liver ascorbic acid concentrations, but liver GLO activity did not differ among groups. Sows were killed at each period; liver GLO activity was constant during gestation but increased postpartum (P < 0.01). Liver ascorbic acid concentration was constant during gestation, except for a decline during late gestation, and increased postpartum (P < 0.05). These results suggest that more ascorbic acid was transferred from the dam to the fetuses as pregnancy advanced, possibly suppressing fetal GLO activity. Thus, fetal liver GLO activity was the primary source of ascorbic acid during early fetal development, but more fetal ascorbic acid was transferred from the dam during later pregnancy. PMID- 11435521 TI - Liver L-gulonolactone oxidase activity and tissue ascorbic acid concentrations in nursing pigs and the effect of various weaning ages. AB - In Experiment 1, we evaluated liver L-gulono-gamma-lactone oxidase (GLO) activity and tissue concentration of ascorbic acid in young pigs from birth to weaning (14 d) and through a 28-d postweaning period; in Experiment 2, we evaluated the effect of three weaning ages on these measurements. Sow colostrum and milk collected in both experiments demonstrated a linear decline (P < 0.01) in ascorbic acid concentration as lactation progressed. In Experiment 1, three pigs were killed at 0, 3, 7, 14, 21, 28, 35 and 42 d of age for determining liver GLO activity and serum and tissue ascorbic acid. Liver GLO activity decreased by 80% from 0 to 3 d of age and remained low until d 14 (weaning). After weaning, liver GLO activity increased linearly (P < 0.01). Tissue ascorbic acid concentrations decreased during the nursing period and again after weaning, but then increased to 42 d of age (P < 0.01). In Experiment 2, pigs were weaned at 10, 17 or 24 d of age. Three pigs from each group were killed at weaning and at each week postweaning until 38 d of age. Liver GLO activity was low during the nursing period but increased linearly (P < 0.01) for each group during the subsequent postweaning period. Pig serum and tissue ascorbic acid concentrations increased postweaning in each group. These results suggest that a factor in sow's milk, possibly ascorbic acid, suppressed liver GLO activity of nursing pigs but upon weaning, liver GLO activity of pigs increased in a linear manner (P < 0.01). PMID- 11435522 TI - Aluminum toxicity alters the regulation of calbindin-D28k protein and mRNA expression in chick intestine. AB - Previous studies have shown that aluminum inhibits vitamin D-dependent calcium absorption. The mechanism involves reduced sensitivity to 1,25 dihydroxycholecalciferol and reduced expression of the calcium transport protein, calbindin-D28k. Reduced expression of calbindin protein may be due to decreased levels of calbindin mRNA. To test this hypothesis, we measured calbindin mRNA levels in chicks fed diets with and without added aluminum. Groups of chicks were fed one of four diets: control, control plus aluminum, low calcium, or low calcium plus aluminum. A fifth group was fed a vitamin D-free diet as a negative control. Calbindin protein was measured by immunoblotting. Serum calcium and inorganic phosphorus were determined. Intestinal mRNA was isolated and assayed by slot-blot hybridization to a fluorescein-conjugated oligonucleotide probe complementary to calbindin-D28k mRNA. Antifluorescein antibodies conjugated to alkaline phosphatase were used to detect hybrids and mRNA levels were quantified by densitometry. Specificity of the probe was verified by Northern analysis. Intestinal calbindin protein was greater in the control plus aluminum group than in controls, but no difference in calbindin mRNA was observed. These changes were associated with small decreases in serum phosphorus and calcium, suggesting a postranscriptional effect of aluminum. Chicks fed the low calcium diet had greater intestinal calbindin protein and mRNA levels relative to the control group in association with a 45% decrease in serum calcium. In contrast, no difference in calbindin protein, and significantly less mRNA were found in the low calcium plus aluminum group compared with controls, despite a decrease in serum calcium similar to that of chicks fed the low calcium diet without aluminum. These results show that in chicks fed a low calcium diet, aluminum intake decreases transcription and/or stability of intestinal calbindin mRNA, and that aluminum may inhibit the expression of vitamin D-dependent genes. PMID- 11435523 TI - Adult cockatiels (Nymphicus hollandicus) metabolically adapt to high protein diets. AB - To determine the ability of cockatiels (Nymphicus hollandicus), a granivorous avian species, to adapt metabolically to high dietary protein levels, adult males (n = 26) were fed isocaloric diets containing 11, 20, 35 or 70% crude protein (CP) for 11 mo. Throughout the trial, body weight and breast muscle weight were maintained by 11, 20 or 70% CP. The 35% CP diet resulted in significantly greater body weight (P < 0.05) and whole-body lipid content (P < 0.05) compared with the 11% CP diet. The 20% CP diet resulted in greater breast muscle mass compared with 70% CP (P < 0.05). Activity of the amino acid catabolic enzymes alanine aminotransferase, aspartate aminotransferase and arginase as well as the gluconeogenic enzyme phosphoenolpyruvate carboxykinase were significantly increased with 70% CP (P < 0.05). Serum essential amino acids, urea and uric acid were also increased with 70% CP (P < 0.05), but the magnitude of their increase was similar to that found in omnivorous chickens fed a similar diet. There was no evidence of visceral gout, articular gout or renal pathology; however liver lesion severity, and specifically liver lipogranuloma severity, was significantly increased above 11% CP (P < 0.05). We conclude that cockatiels are able to up regulate enzymes for amino acid catabolism as well as mechanisms for nitrogen excretion in response to high dietary protein levels, and that high dietary protein levels are not associated with kidney dysfunction in this avian species. PMID- 11435524 TI - R3230AC rat mammary tumor and dietary long-chain (n-3) fatty acids change immune cell composition and function during mitogen activation. AB - Because anticancer immunity declines progressively with tumor growth, a major focus of current research in tumor immunology is the development of means to stimulate the host immune system. This study determined the effects of dietary long-chain (n-3) fatty acids and tumor burden on immune cell phospholipid composition and membrane-mediated immune defense in rats implanted with the R3230AC mammary adenocarcinoma. Fischer 344 rats (145 +/- 2 g) were fed one of two semipurified diets (20 g/100 g fat) for 21 d before and 17 d after tumor implantation. Diets provided long-chain (n-3) fatty acids at 0 or 50 g/kg of total fat. Mammary tumor growth was 31% lower (P = 0.1) in rats fed long-chain (n 3) fatty acids. Dietary long-chain (n-3) fatty acids had beneficial effects on several host immune defenses, including activation of CD8(+) T cells and type-1 cytokine (interferon-gamma and tumor necrosis factor-alpha) production (P < 0.05). Upregulated immune function in tumor-bearing rats fed the high (n-3) diet occurred concurrently with specific changes in the major membrane phospholipids phosphatidylcholine and phosphatidylethanolamine in high (n-3)-fed rats. Because membrane composition plays a critical role in immune function, additional work is needed to determine the relationship between alterations in the phospholipid composition of immune cells during cancer and subsequent upregulation of host defense in the tumor-bearing state. PMID- 11435525 TI - Iron deficiency alters the progression of mitogen-treated murine splenic lymphocytes through the cell cycle. AB - The influence of iron deficiency on the progression of mitogen-treated splenic lymphocytes through the cell cycle was studied in 16 control, 16 pair-fed, 15 iron-deficient (ID) and 16 ID mice that were repleted for up to 3 d (R3). The test and control diets differed only in iron concentrations (0.09 vs. 0.9 mmol/kg). When mice were killed (68 d of feeding), the hemoglobin concentration and liver iron stores of ID and R3 mice were <50% those of control mice (P < 0.05). Iron deficiency did not reduce the percentage of CD3(+) cells, but decreased CD3(+) cells/mg spleen (P < 0.05). In concanavalin A-treated and nonactivated cultures, there were no significant differences among groups in the percentages of cells in resting phase of the cell cycle (G0) to cell cycle initiation phase (G1), DNA synthesis phase (S) and exit from the S phase (G2) to mitosis phase (M) phases. In anti-CD3 and anti-CD3/anti-CD28-treated cultures, higher percentages of lymphocytes from ID and R3 mice than those from control and pair-fed mice were in the G0--G1 phase (P < 0.05). Conversely, lower percentages of activated cells from ID and R mice than those from control and pair-fed mice were in S and G2--M phases (P < 0.05). Incubation of lymphocytes with mitogens decreased the percentages of cells in G0--G1 phase from 90% to 80% in control and pair-fed but not in ID and R3 mice (P < 0.05). In activated cells, indices of iron status negatively correlated with the percentages of cells in G0--G1 (r = 0.306 to -0.597) but positively with those in S (r = 0.166--0.511) and G2--M phases (r = 0.265-0.59; P < 0.05). Data suggest that altered cell cycle progression likely contributes to impaired lymphocyte proliferation usually associated with iron deficiency. PMID- 11435526 TI - Green tea polyphenol extract attenuates inflammation in interleukin-2-deficient mice, a model of autoimmunity. AB - Green tea polyphenols (GrTP) have been previously shown to decrease endotoxin induced tumor necrosis factor-alpha production and lethality in mice. Our present studies demonstrate that GrTP inhibit inflammatory responses and may be useful in treating chronic inflammatory states, such as inflammatory bowel disease. In this preliminary study, we examined whether GrTP decrease disease activity in interleukin-2-deficient (IL-2(-/-) mice. Eight-week old IL-2(-/-) C57BL/6J mice who were fed nonpurified diet were randomly assigned to receive water with GrTP (5 g/L) or water alone (control) for up to 6 wk. After 1 wk, explant colon and lamina propria lymphocyte (LPL) cultures were established from a subgroup of mice and supernatants collected. Culture supernatants from GrTP-treated mice showed decreased spontaneous interferon-gamma and tumor necrosis factor-alpha secretion compared with that of controls. At 6 wk, the GrTP group had less severe colitis as demonstrated by lower histologic scores and wet colon weights. This was associated with lower plasma levels of serum amyloid A, increased weight gain and improved hematocrits. These results show that GrTP attenuated inflammation in IL 2(-/-) mice and suggest a role for GrTP in treating chronic inflammatory diseases such as inflammatory bowel disease. PMID- 11435527 TI - Low serum and red blood cell folate are moderately, but nonsignificantly associated with increased risk of invasive cervical cancer in U.S. women. AB - Previous observational epidemiologic studies of folate and cervical cancer, as well as folate supplementation trials for cervical dysplasia, have produced mixed results. We examined the relationship between serum and RBC folate and incident invasive cervical cancer in a large, multicenter, community-based case-control study. Detailed in-person interviews were conducted, and blood was drawn at least 6 mo after completion of cancer treatment from 51% of cases and 68% of controls who were interviewed. Blood folate was measured with both microbiologic and radiobinding assays. Included in the final analyses were 183 cases and 540 controls. Logistic regression was used to control for all accepted risk factors, including age, sexual behavior, smoking, oral contraceptive use, Papanicolaou smear history and human papillomavirus (HPV)-16 serology. For all four folate measures, the geometric mean in cases was lower than in controls (e.g., 11.6 vs. 13.0 nmol/L, P < 0.01 for the serum radiobinding assay). Folate measures using microbiologic and radiobinding assays were correlated (serum: r = 0.90; RBC: r = 0.77). For serum folate, multivariate-adjusted odds ratios (OR) in the lowest vs. highest quartile were 1.3 [95% confidence interval (CI) = 0.8--2.9] and 1.6 (0.9- 2.9), using the microbiologic and radiobinding assays, respectively. For RBC folate, comparable OR were 1.2 (0.6--2.2) and 1.5 (0.8--2.7). Similar risks were obtained when restricting analyses to subjects with a history of HPV infection. Thus, low serum and RBC folate were each moderately, but nonsignificantly, associated with increased invasive cervical cancer risk. These findings support a role for one-carbon metabolism in the etiology of cervical cancer. PMID- 11435528 TI - Body composition in 70-year-old adults responds to dietary beta-hydroxy-beta methylbutyrate similarly to that of young adults. AB - Studies in young adults have demonstrated that beta-hydroxy-beta-methylbutyrate (HMB) can increase gains in strength and fat-free mass during a progressive resistance-training program. The purpose of this study was to determine whether HMB would similarly benefit 70-y-old adults undergoing a 5 d/wk exercise program. Thirty-one men (n = 15) and women (n = 16) (70 +/- 1 y) were randomly assigned in a double-blind study to receive either capsules containing a placebo or Ca-HMB (3 g/d) for the 8-wk study. Skin fold estimations of body composition as well as computerized tomography (CT) and dual X-ray absorptiometry (DXA) scans were measured before the study and immediately after the 8-wk training program. HMB supplementation tended to increase fat-free mass gain (HMB, 0.8 +/- 0.4 kg; placebo, -0.2 +/- 0.3 kg; treatment x time, P = 0.08). Furthermore, HMB supplementation increased the percentage of body fat loss (skin fold: HMB, -0.66 +/- 0.23%; placebo, -0.03 +/- 0.21%; P = 0.05) compared with the placebo group. CT scans also indicated a greater decrease in the percentage of body fat with HMB supplementation (P < 0.05). In conclusion, changes in body composition can be accomplished in 70-y-old adults participating in a strength training program, as previously demonstrated in young adults, when HMB is supplemented daily. PMID- 11435529 TI - Nutritional status assessment in semiclosed environments: ground-based and space flight studies in humans. AB - Adequate nutrition is critical during long-term spaceflight, as is the ability to easily monitor dietary intake. A comprehensive nutritional status assessment profile was designed for use before, during and after flight. It included assessment of both dietary intake and biochemical markers of nutritional status. A spaceflight food-frequency questionnaire (FFQ) was developed to evaluate intake of key nutrients during spaceflight. The nutritional status assessment protocol was evaluated during two ground-based closed-chamber studies (60 and 91 d; n = 4/study), and was implemented for two astronauts during 4-mo stays on the Mir space station. Ground-based studies indicated that the FFQ, administered daily or weekly, adequately estimated intake of key nutrients. Chamber subjects maintained prechamber energy intake and body weight. Astronauts tended to eat 40--50% of WHO predicted energy requirements, and lost >10% of preflight body mass. Serum ferritin levels were lower after the chamber stays, despite adequate iron intake. Red blood cell folate concentrations were increased after the chamber studies. Vitamin D stores were decreased by > 40% on chamber egress and after spaceflight. Mir crew members had decreased levels of most nutritional indices, but these are difficult to interpret given the insufficient energy intake and loss of body mass. Spaceflight food systems can provide adequate intake of macronutrients, although, as expected, micronutrient intake is a concern for any closed or semiclosed food system. These data demonstrate the utility and importance of nutritional status assessment during spaceflight and of the FFQ during extended duration spaceflight. PMID- 11435530 TI - Does dietary protein in early life affect the development of adiposity in mammals? AB - This article examines the proposition that dietary protein in pre- and early postnatal life influences the development of adiposity in later life. In rodents, low protein intake during gestation can result in low birth weight and subsequently leads to various metabolic disturbances in adulthood, such as high blood pressure, impaired glucose tolerance and insulin resistance. The few controlled studies conducted in animals suggest that high protein or energy intake during gestation leads to low birth weights. Observational studies in humans have been inconclusive in establishing a relationship between dietary protein intake in pregnancy and effects on birth weight and adiposity of the offspring later in life. There is only weak epidemiological evidence linking high protein intake during early childhood and the development of obesity. By contrast, studies in domestic animals have found that higher levels of protein intake are often associated with lower rates of fat accretion. Additional studies are proposed to explore claims linking protein nutrition in early life to the postnatal development of obesity and disease in humans. PMID- 11435531 TI - Differences between humans and mice in efficacy of the body fat lowering effect of conjugated linoleic acid: role of metabolic rate. PMID- 11435533 TI - Appraising decision and cost-effectiveness analyses. PMID- 11435534 TI - The 2001 Charles T. Dotter lecture: understanding vascular devices at the molecular level is the key to progress. PMID- 11435535 TI - Thrombolytic and antiplatelet therapy in peripheral vascular disease with use of reteplase and/or abciximab. The SCVIR Consultants' Conference; May 22, 2000; Orlando, FL. Society for Cardiovascular and Interventional Radiology. PMID- 11435536 TI - Transcatheter interventions for the treatment of peripheral atherosclerotic lesions: part II. AB - Transcatheter endovascular procedures are increasingly used to treat symptomatic peripheral atherosclerosis. This second part of a two-part review assesses the existing supportive evidence for the application of recently introduced transcatheter treatments for lesions that cause cerebrovascular ischemia and stroke. Studies were identified via MEDLINE (January 1993 through April 1999) and reference lists of identified articles. When multicenter prospective randomized trials or other high-quality studies were unavailable, studies with at least 50 patients per treated group and a minimum follow-up duration of 6 months were included. For each application, the authors assessed the quality of evidence (efficacy, safety, and, where available, cost-effectiveness) and made recommendations with appropriate caveats. Although recommendations based on proven efficacy and cost-effectiveness cannot be made in general, the use of transcatheter therapies can be supported in specific circumstances based on expected reduction in procedure-related morbidity and/or mortality. It is hoped that the identification of deficiencies in the literature will inform and inspire critically needed research in this area. PMID- 11435537 TI - Proximity arteriography: cost-effectiveness in asymptomatic penetrating extremity trauma. AB - PURPOSE: Many urban trauma centers have abandoned proximity arteriography, which is defined as exclusion arteriography used to evaluate the asymptomatic patient with penetrating extremity trauma near major arteries. However, decision analysis has not been applied to study proximity arteriography. MATERIALS AND METHODS: The cost-effectiveness of proximity arteriography was examined by creating a decision model that compared arteriography with observation after patient examination in the trauma unit. The model used predominantly literature-derived estimates for input variables and outcomes. The authors retrospectively reviewed arteriograms for 1 year to identify major occult injuries (requiring intervention) at their institution. After a resource-based cost analysis from the taxpayers' perspective, the cost-effectiveness ratio was calculated (incremental cost per quality-adjusted life year [QALY] gained) for proximity arteriography. RESULTS: For proximity trauma, arteriography is a dominant strategy (more effective and costs less) at a prevalence of major occult injury of 5.5% or more. Observation is a dominant strategy if the prevalence is from 0% to 0.5% or 2.0%, depending on arteriography complication assumptions. In between, arteriography is cost effective, with a ratio of $12,100 per QALY at 2.5% prevalence (base case). Besides prevalence of occult injury, the model is sensitive to outcome assumptions for occult injuries. CONCLUSION: Proximity arteriography is a cost effective procedure if major occult injuries equal or exceed 1.0%. PMID- 11435538 TI - Cost-effectiveness of percutaneous radiofrequency ablation for malignant hepatic neoplasms. AB - PURPOSE: Percutaneous radiofrequency (RF) ablation is a promising technique for the treatment of hepatic malignancies. However, its cost-effectiveness has not been established. The purpose of this study is to determine the cost effectiveness of RF ablation compared to palliative care in the treatment of hepatocellular cancer and colorectal liver metastases. This study also seeks to evaluate the effects of transition from traditional to newly implemented prospective outpatient reimbursement mechanisms on RF ablation cost effectiveness. MATERIALS AND METHODS: The marginal direct costs of a percutaneous RF ablation treatment strategy were compared to palliative care over a range of survival benefits with use of a cost-effectiveness model built from the perspective of the payer. Variables used in the model, including complication rates and procedure efficacy, were obtained from the literature and the authors' experience with 46 consecutive patients. RESULTS: The cost-effectiveness of a standardized percutaneous RF ablation treatment strategy compared to palliative care was $20,424, $11,407, $5,034, and $3,492, respectively, per life-year (LY) gained when marginal median survival conferred by RF ablation is 6 months, 1 year, 3 years, and 5 years. The RF ablation treatment strategy would be required to generate 6.14, 2.26, and 1.10 months of marginal median survival benefit to achieve strict ($20,000/LY gained), moderate ($50,000/LY gained), and generous ($100,000/LY gained) cost-effectiveness thresholds. Cost-effectiveness was sensitive to the number of lifetime treatments, hours of observation time, frequency of follow-up evaluations, cost of abdominal computed tomography, and decision to perform RF ablation as an inpatient or outpatient. CONCLUSION: Percutaneous RF ablation is a cost-effective treatment strategy compared to palliative care and has likely already achieved the survival benefit required to meet even a strict cost-effectiveness criterion. Dependence on reimbursement mechanism highlights the importance of concordance between policy and RF ablation technology. The results of this study allow flexible application of cost effectiveness data despite current uncertainties in treatment and survival data and heterogeneity in treatment populations. PMID- 11435539 TI - Treatment of chronic aortic dissection by transluminal endovascular stent-graft placement: preliminary results. AB - PURPOSE: To investigate efficacy of stent-graft repair for the treatment of patients with chronic aortic dissection. MATERIALS AND METHODS: Fifteen patients with chronic aortic dissection were treated with endovascular stent-grafts. Entry tears were located in the descending thoracic aorta in all patients. The mean maximum diameter of the descending thoracic aorta was 47 mm +/- 8. The mean diameter of the true lumen at the same level was 20 mm +/- 5. The mean interval between diagnosis and stent-graft procedure was 32 months +/- 91. Stent-grafts were fabricated from expanded polytetrafluoroethylene and Z-stents. RESULTS: Stent-grafts were placed successfully in all patients. Two stent-grafts were required in one patient. Entry closure and thrombosis of the false lumen of the descending thoracic aorta were also achieved in all patients. No procedure related complications were observed except for postimplantation syndrome, including fever and leukocytosis. The diameter of the true lumen was significantly increased (mean, 31 mm +/- 6) at the level of the descending thoracic aorta (P <.01) and the diameter of the aorta was significantly decreased (mean, 44 mm +/- 8) at the same level (P <.01). There were no deaths and no instances of aortic rupture during the subsequent average follow-up period of 24 months. Secondary stent-graft procedures were required to treat the abdominal component of dissection during follow-up in one patient. CONCLUSIONS: Stent-graft repair of chronic aortic dissection is a safe and effective method and may be an alternative to surgical graft replacement in selected patients. However, further evaluation is mandatory before this method is widely employed. PMID- 11435540 TI - Utility of nonselective abdominal aortography in demonstrating ovarian artery collaterals in patients undergoing uterine artery embolization for fibroids. AB - PURPOSE: To review the incidence of ovarian collateral supply to uterine fibroids as demonstrated by nonselective abdominal aortography before uterine artery embolization (UAE) and to evaluate the effect of such visualization on interventional management. MATERIALS AND METHODS: The aortograms of 51 consecutive patients (mean age, 42.4 y; range, 30--53 y) undergoing UAE for symptomatic uterine fibroids were reviewed retrospectively for the visualization of ovarian arteries extending into the pelvis. If ovarian arteries were visualized, their size relative to the ipsilateral external iliac artery was measured. Arteries believed large enough to represent a significant blood supply to the uterine fibroids were further evaluated after UAE to determine whether flow persisted. RESULTS: In 13 of 51 patients (25%), a total of 18 ovarian arteries were identified. They were bilateral in five patients and unilateral in eight. Their sizes relative to the ipsilateral external iliac artery ranged from 8% to 57% (mean, 26%). Eight ovarian arteries with a relative size > or = 25% were further evaluated. Five of the eight (62.5%) were not visible after UAE. Of the three persistent ovarian collateral arteries, two were successfully embolized. The patient with the untreated collateral artery experienced persistent menorrhagia. CONCLUSION: Preembolization aortography with the catheter tip at level of the renal arteries demonstrated ovarian collateral arteries in 25% of patients with uterine fibroids. However, their detection influenced treatment in only 6% of the reported cases. PMID- 11435541 TI - Long-term prognosis of patients undergoing transcatheter arterial chemoembolization for unresectable hepatocellular carcinoma: comparison of cisplatin lipiodol suspension and doxorubicin hydrochloride emulsion. AB - PURPOSE: To evaluate long-term prognosis of transcatheter arterial chemoembolization (TACE) with use of cisplatin (CDDP) lipiodol (LPD) suspension (CDDP/LPD) compared with that with use of doxorubicin hydrochloride (ADM) LPD emulsion (ADM/LPD) in patients with unresectable hepatocellular carcinoma (HCC). MATERIALS AND METHODS: One hundred eight patients were treated with use of CDDP/LPD and 26 were treated with use of ADM/LPD. Survival rates and frequency of side effects and complications in the CDDP/LPD group were compared with those in the ADM/LPD group. RESULTS: CDDP/LPD was given at a dose of 15-70 mg (mean dose, 41 mg), whereas ADM/LPD was given at a dose of 20-100 mg (mean dose, 57 mg) throughout the study period. The survival rates in the CDDP/LPD group were 81% at 1 year, 41% at 3 years, 19% at 5 years, and 13% at 7 years, whereas those in the ADM/LPD group were 67% at 1 year, 18% at 3 years, and 0% at 5 years. The CDDP/LPD group showed significantly better survival than the ADM/LPD group (P <.05). In the CDDP/LPD group, there was a significant prolongation of survival in patients with monofocal HCC (P <.05) and patients with HCC assessed as an almost complete LPD accumulation (P <.05). There were no significant differences in survival rates in the ADM/LPD group according to tumor size and number of tumors. Hepatic failure was observed in 8% of all procedures and was not different between the two therapeutic groups. Renal dysfunction was observed in 2% of all treatments involving CDDP/LPD, and it resolved spontaneously with appropriate medications. CONCLUSIONS: TACE with use of low-dose CDDP was efficacious for unresectable HCC and had few complications. TACE with use of CDDP may contribute to prolongation of the life span of patients with HCC versus TACE with use of ADM. PMID- 11435542 TI - Irradiated versus nonirradiated endothelial cells: effect on proliferation of vascular smooth muscle cells. AB - PURPOSE: Endovascular radiation therapy is a promising strategy for the prevention of restenosis. Radiation prevents proliferation of vascular smooth muscle cells, thereby reducing the incidence of restenosis, but may also affect the remaining endothelial cells. For this reason, a comparison was made between irradiated and nonirradiated endothelial cells and their effects on the proliferation of vascular smooth muscle cells in a coculture system was evaluated. MATERIALS AND METHODS: A coculture system was used, in which both endothelial cells and vascular smooth muscle cells were grown on opposite sides of a semipermeable membrane. After a period of growth arrest, the proliferation of vascular smooth muscle cells was measured during four subsequent days. RESULTS: The presence of endothelial cells stimulated the proliferation of vascular smooth muscle cells during the first days of analysis but had an inhibitory effect during the subsequent days (P <.5). gamma-irradiation of endothelial cells resulted in a complete blockage of the proliferation of these cells. However, irradiated endothelial cells affected the proliferation of vascular smooth muscle cells in coculture in a fashion comparable to nonirradiated endothelial cells (P >.5). CONCLUSION: The results suggest that, in endovascular radiation therapy, irradiation of endothelial cells does not change their effects on the proliferative behavior of vascular smooth muscle cells. PMID- 11435543 TI - Renal artery embolization with diluted hot contrast medium: an experimental study. AB - PURPOSE: To evaluate the safety and efficacy of diluted hot contrast medium to embolize the renal arteries in a canine model. MATERIALS AND METHODS: Transcatheter embolization was performed in 15 dogs (five dogs in each phase: I, II, and control). The diluted hot contrast medium was heated to 90 degrees C--100 degrees C and 30--60 mL were injected into a renal artery of each dog in phase I (optimization studies) and II (efficacy studies). In the control group, balloon inflation was performed without injection of diluted hot contrast medium. The temperature measurement was performed on one dog in phase I by temperature probe. Follow-up angiography was performed immediately (n = 15), at 1 week (n = 9), at 1 month (n = 9), and after 3 months (n = 5). All dogs were killed and kidneys were removed for histologic examination. RESULTS: The procedure was performed successfully in all dogs. Follow-up angiography showed complete obstruction of the injected renal arteries in phases I and II. The microscopic findings showed acute full-thickness necrosis of the renal cortex and renal artery in phase I, and complete coagulative necrosis of the cortex, medulla, and intrarenal blood vessels in phase II. There were no histologic changes of renal veins, inferior vena cava (IVC), and aorta. There were nonspecific findings in the control group. CONCLUSION: This preliminary experimental study suggests that diluted hot contrast medium may be a safe agent for renal ablation without thrombus formation in the renal vein or IVC. PMID- 11435544 TI - New embolization coil containing a nitinol wire core: preliminary in vitro and in vivo experiences. AB - PURPOSE: To improve the stability of the standard stainless steel embolization coil by adding a nitinol wire core. MATERIALS AND METHODS: With use of one coil with a nitinol wire core and one without one, stability and resistance to elongation were measured in vitro. Thirty-one factory-made stainless-steel macrocoils equipped with preshaped nitinol wire cores were acutely deployed into branches of the superior mesenteric artery (SMA) in five pigs (part I of the study). Fifteen homemade retrievable coils with thermal shape memory (TSM) nitinol wire cores were acutely deployed in the abdominal aortae or inferior venae cavae of four pigs (part II). Coils with a superelastic (SE) nitinol wire core (n = 9), a TSM nitinel wire core (n = 5), and without a core (n = 5) were compared in carotid embolization (part III). RESULTS: In vitro, the expansile strength of the reinforced coils was significantly greater and elongation was significantly less than the standard coils (P <.01). In part I, coils were easily deployed via diagnostic catheters. In part II, ability to reposition the coils facilitated optimal coil configuration, which resulted in effective self anchoring and occlusion. In part III, no coils with a SE core migrated, whereas one of five with a TSM core and three of five without reinforcement migrated immediately. The core significantly increased coil stability and postplacement configuration (P <.01). CONCLUSIONS: Addition of a nitinol wire core increases the intravascular stability of the standard embolization coil by significantly enhancing expansile force and postplacement configuration. The wire core does not adversely affect the handling of the coil. The ability to reposition/retrieve the coil is a desirable feature. PMID- 11435545 TI - Portal-systemic myelopathy after transjugular intrahepatic portosystemic shunt creation: report of four cases. AB - The authors attempted to describe the clinical manifestations of portal-systemic myelopathy (PSM) after transjugular intrahepatic portosystemic shunt (TIPS) creation. PSM was developed in four of 212 (1.89%) patients who underwent TIPS procedures in our hospital. Three men and one woman, ranging in age from 41 to 56 years, with a history of posthepatitis cirrhosis and recurrent bleeding from gastroesophageal varices had intrahepatic shunts created with 10-mm-diameter Wallstents. Shunt patency was confirmed by color Doppler ultrasonography (US) in each patient after TIPS creation. Progressive spastic paraparesis involving the lower extremities occurred between 5 weeks and 5 months after TIPS creation in the four patients. Neurologic examination showed evidence of spasticity in all cases, with ankle clonus, extensor plantar responses, and lower extremity hyperreflexia. All sensory modalities remained intact. Cytologic examination of cerebrospinal fluid from each patient was normal. There was no evidence of spinal cord compression on the imaging studies. PSM is a rare syndrome that includes spastic paraparesis with intact sensation. Initially noted in patients who have undergone surgical placement of a portacaval shunt, it also may occur after TIPS creation. PMID- 11435546 TI - Portal vein embolization with use of polyvinyl alcohol particles. AB - Preoperative portal vein embolization has been used as method of inducing compensatory hypertrophy in the future remnant liver since it was first described in the late 1980s. Many different vascular embolic agents have been successfully used for this procedure, and there is no general consensus regarding which is the best agent. Polyvinyl alcohol (PVA) particles commonly used for arterial embolization come in many sizes, are readily available, and are easy to administer via conventional catheters. We describe an easy, safe, and effective method of the use of PVA particles for portal vein embolization. PMID- 11435547 TI - Successful treatment of TIPS-induced hepatic failure with an hourglass stent graft: a simple new technique for reducing shunt flow. AB - A simple new technique for creating an hourglass reducing stent-graft by means of targeted balloon expansion of each end of a commercially available device is presented. A 67-year-old man with progressive hepatic failure after TIPS creation was treated. A 48-mm-long Jostent stent-graft was placed inside the TIPS through a 10-F introducer and fully expanded only at the proximal and distal ends, resulting in an hourglass shape. Immediate increase of portal pressure was achieved, followed by complete clinical recovery. Advantages of covered versus bare reducing stents are discussed. PMID- 11435548 TI - Real-time I-MR-imaging--guided cryoablation of uterine fibroids. AB - In an ongoing prospective clinical trial, two patients with large, symptomatic uterine fibroids were treated with percutaneous magnetic resonance (MR)-imaging- guided cryoablation. Patients were followed with serial MR imaging studies and clinical evaluation according to Institutional Review Board protocol. Short-term imaging and clinical data were collected to assess the effects of the therapy. Both patients confirmed a significant reduction or elimination of their symptoms at 2 weeks after cryoablation. Measurements obtained from MR imaging performed after cryoablation documented a 65% and 53% reduction in each patient's fibroid volume at 8 weeks. Early clinical and imaging results confirm MR-imaging--guided cryoablation was effective at reducing or relieving these two patients' symptoms related to uterine fibroids and post-cryoablation imaging confirmed a significant reduction in fibroid volume. PMID- 11435549 TI - Re: Contrast media-induced nephrotoxicity: identification of patients at risk and algorithms for prevention. PMID- 11435550 TI - Re: Endovascular treatment of acute carotid blow-out syndrome. PMID- 11435551 TI - Fatal pulmonary embolus after TIPS revision. PMID- 11435552 TI - Destabilization of the retinoblastoma tumor suppressor by human papillomavirus type 16 E7 is not sufficient to overcome cell cycle arrest in human keratinocytes. AB - The E7 oncoprotein of human papillomavirus type 16 promotes cell proliferation in the presence of antiproliferative signals. Mutagenesis of E7 has revealed that this activity requires three regions, conserved regions 1 and 2 and a C-terminal zinc finger. Binding to the retinoblastoma tumor repressor (Rb) through an LxCxE motif in conserved region 2 is necessary, but not sufficient, for E7 to induce proliferation. We tested the hypothesis that binding to Rb is not sufficient because conserved region 1 and/or the C terminus are required for E7 to functionally inactivate Rb and thus induce proliferation. One mechanism proposed for how E7 inactivates Rb is by blocking Rb-E2F binding. Either conserved region 1 or the C terminus was necessary, in combination with the LxCxE motif, for E7 to block Rb-E2F binding in vitro. While all full-length E7 proteins with mutations outside of the LxCxE motif inhibited Rb-E2F binding, some failed to abrogate cell cycle arrest, demonstrating that blocking Rb-E2F binding is not sufficient for abrogating antiproliferative signals. Another mechanism proposed for how E7 inactivates Rb is by promoting the destabilization of Rb protein. Mutations in conserved region 1 or the LxCxE motif prevented E7 from reducing the half-life of Rb. Though no specific C-terminal residues of E7 were essential for destabilizing Rb, a novel class of mutations that uncouple the destabilization of Rb from the deregulation of keratinocyte proliferation was discovered. Destabilization of Rb correlated with the abrogation of Rb-induced quiescence but was not sufficient for overriding DNA damage-induced cell cycle arrest or for increasing keratinocyte life span. Finally, the same regions of E7 required for destabilizing Rb were required for reducing p107 and p130 levels. Together, these results suggest that inactivation of all three Rb family members is not sufficient to deregulate keratinocyte cell cycle control. PMID- 11435553 TI - OX40 stimulation by gp34/OX40 ligand enhances productive human immunodeficiency virus type 1 infection. AB - OX40 is a member of the tumor necrosis factor (TNF) receptor superfamily and known to be an important costimulatory molecule expressed on activated T cells. To investigate the role of costimulation of OX40 in human immunodeficiency virus type 1 (HIV-1) infection by its natural ligand, gp34, the OX40-transfected ACH-2 cell line, ACH-2/OX40, chronically infected with HIV-1, was cocultured with paraformaldehyde (PFA)-fixed gp34-transfected mouse cell line, SV-T2/gp34. The results showed that HIV-1 production was strongly induced. This was followed by apparent apoptosis, and both processes were specifically inhibited by the gp34 specific neutralizing monoclonal antibody 5A8. Endogenous TNF alpha (TNF-alpha) and TNF-beta production were not involved in the enhanced HIV-1 production. Furthermore, enhanced HIV-1 transcription in gp34-stimulated ACH-2/OX40 cells was dependent on the kappa B site of the HIV-1 long terminal repeat, and the OX40 gp34 interaction activated NF-kappa B consisting of p50 and p65 subunits. When primary activated CD4(+) T cells acutely infected with HIV-1(NL4-3) (CXCR4-using T-cell-line-tropic) were cocultured with PFA-fixed gp34(+) human T-cell leukemia virus type 1-bearing MT-2 cells or SV-T2/gp34 cells, HIV-1 production was also markedly enhanced. The enhancement was again significantly inhibited by 5A8. The present study first shows that OX40-gp34 interaction stimulates HIV-1 expression and suggests that OX40 triggering by gp34 may play an important role in enhancing HIV-1 production in both acutely and latently infected CD4(+) T cells in vivo. PMID- 11435554 TI - African swine fever virus structural protein pE120R is essential for virus transport from assembly sites to plasma membrane but not for infectivity. AB - This report examines the role of African swine fever virus (ASFV) structural protein pE120R in virus replication. Immunoelectron microscopy revealed that protein pE120R localizes at the surface of the intracellular virions. Consistent with this, coimmunoprecipitation assays showed that protein pE120R binds to the major capsid protein p72. Moreover, it was found that, in cells infected with an ASFV recombinant that inducibly expresses protein p72, the incorporation of pE120R into the virus particle is dependent on p72 expression. Protein pE120R was also studied using an ASFV recombinant in which E120R gene expression is regulated by the Escherichia coli lac repressor-operator system. In the absence of inducer, pE120R expression was reduced about 100-fold compared to that obtained with the parental virus or the recombinant virus grown under permissive conditions. One-step virus growth curves showed that, under conditions that repress pE120R expression, the titer of intracellular progeny was similar to the total virus yield obtained under permissive conditions, whereas the extracellular virus yield was about 100-fold lower than in control infections. Immunofluorescence and electron microscopy demonstrated that, under restrictive conditions, intracellular mature virions are properly assembled but remain confined to the replication areas. Altogether, these results indicate that pE120R is necessary for virus dissemination but not for virus infectivity. The data also suggest that protein pE120R might be involved in the microtubule-mediated transport of ASFV particles from the viral factories to the plasma membrane. PMID- 11435555 TI - Frameshift signal transplantation and the unambiguous analysis of mutations in the yeast retrotransposon Ty1 Gag-Pol overlap region. AB - The yeast retrotransposon Ty1 encodes a 7-nucleotide RNA sequence that directs a programmed, +1 ribosomal frameshifting event required for Gag-Pol translation and retrotransposition. We report mutations that block frameshifting, which can be suppressed in cis by "transplanting" the frameshift signal to a position upstream of its native location. These "frameshift transplant" mutants transpose with only a modest decrease in efficiency, suggesting that the location of the frameshift signal in a functional Ty1 element may vary. The genomic architecture of Ty1 is such that Gag, Ty1 PR (PR), and the Gag-derived p4 peptide share a common sequence. The functional independence of the movement of the frameshift signal to a new location within the Ty1 element is used to unambiguously attribute the effect of mutations deleterious to transposition in this region of overlapping coding sequences to effects on the Ty1 (PR). This work defines the amino terminus of the Ty1 PR and introduces a new technique for studying viral genome organization. PMID- 11435556 TI - Molecular basis for cell tropism of CXCR4-dependent human immunodeficiency virus type 1 isolates. AB - Laboratory isolates of human immunodeficiency virus type 1 (HIV-1) that utilize CXCR4 as a coreceptor infect primary human macrophages inefficiently even though these express a low but detectable level of cell surface CXCR4. In contrast, infection of primary macrophages by primary CXCR4-tropic HIV-1 isolates is readily detectable. Here, we provide evidence suggesting that this difference in cell tropism results from a higher requirement for cell surface CXCR4 for infection by laboratory HIV-1 isolates. Transfected COS7 cells that express a high level of CD4 but a low level of CXCR4 were infected significantly more efficiently by two primary CXCR4-tropic HIV-1 isolates compared to the prototypic laboratory HIV-1 isolate IIIB. More importantly, overexpression of either wild type or signaling-defective CXCR4 on primary macrophages dramatically enhanced the efficiency of infection by the laboratory HIV-1 isolate yet only modestly enhanced infection by either primary CXCR4-tropic virus. Overexpression of CD4 had, in contrast, only a limited effect on macrophage infection by the laboratory HIV-1, although infection by the primary isolates was markedly enhanced. We therefore conclude that the laboratory CXCR4-tropic HIV-1 isolate exhibits a significantly higher CXCR4 requirement for efficient infection than do the primary CXCR4-tropic isolates and that this difference can explain the poor ability of the laboratory HIV-1 isolate to replicate in primary macrophages. More generally, we propose that the cell tropisms displayed by different strains of HIV-1 in culture can largely be explained on the basis of differential requirements for cell surface CD4 and/or coreceptor expression levels. PMID- 11435557 TI - DNA binding by Kaposi's sarcoma-associated herpesvirus lytic switch protein is necessary for transcriptional activation of two viral delayed early promoters. AB - Kaposi's sarcoma-associated herpesvirus (KSHV; also known as human herpesvirus-8) establishes latent and lytic infections in both lymphoid and endothelial cells and has been associated with diseases of both cell types. The KSHV open reading frame 50 (ORF50) protein is a transcriptional activator that plays a central role in the reactivation of lytic viral replication from latency. Here we identify and characterize a DNA binding site for the ORF50 protein that is shared by the promoters of two delayed early genes (ORF57 and K-bZIP). Transfer of this element to heterologous promoters confers on them high-level responsiveness to ORF50, indicating that the element is both necessary and sufficient for activation. The element consists of a conserved 12-bp palindromic sequence and less conserved sequences immediately 3' to it. Mutational analysis reveals that sequences within the palindrome are critical for binding and activation by ORF50, but the presence of a palindrome itself is not absolutely required. The 3' flanking sequences also play a critical role in DNA binding and transactivation. The strong concordance of DNA binding in vitro with transcriptional activation in vivo strongly implies that sequence-specific DNA binding is necessary for ORF50-mediated activation through this element. Expression of truncated versions of the ORF50 protein reveals that DNA binding is mediated by the amino-terminal 272 amino acids of the polypeptide. PMID- 11435558 TI - Longer and shorter forms of Sendai virus C proteins play different roles in modulating the cellular antiviral response. AB - The Sendai virus (SeV) C gene codes for a nested set of four C proteins that carry out several functions, including the modulation of viral RNA synthesis and countering of the cellular antiviral response. Using mutant C genes (and in particular a C gene with a deletion of six amino acids present only in the larger pair of C proteins) and recombinant SeV carrying these mutant C genes, we find that the nested set of C proteins carry out a nested set of functions. All of the C proteins interdict interferon (IFN) signaling to IFN-stimulated genes (ISGs) and prevent pY701-Stat1 formation. However, only the larger C proteins can induce STAT1 instability, prevent IFN from inducing an antiviral state, or prevent programmed cell death. Remarkably, interdiction of IFN signaling to ISGs and the absence of pY701-Stat1 formation did not prevent IFN-alpha from inducing an anti Vesicular stomatitis virus (VSV) state. It is possible that IFN-alpha signaling to induce an anti-VSV state can occur independently of the well-established Jak/Stat/ISGF3 pathway and that it is this parallel pathway that is targeted by the longer C proteins. PMID- 11435559 TI - Origin binding protein-containing protein-DNA complex formation at herpes simplex virus type 1 oriS: role in oriS-dependent DNA replication. AB - Initiation of herpes simplex virus type 1 (HSV-1) DNA replication during productive infection of fibroblasts and epithelial cells requires attachment of the origin binding protein (OBP), one of seven essential virus-encoded DNA replication proteins, to specific sequences within the two viral origins, oriL and oriS. Whether initiation of DNA replication during reactivation of HSV-1 from neuronal latency also requires OBP is not known. A truncated protein, consisting of the C-terminal 487 amino acids of OBP, termed OBPC, is the product of the HSV UL8.5 gene and binds to origin sequences, although OBPC's role in HSV DNA replication is not yet clear. To characterize protein-DNA complex formation at oriS in cells of neural and nonneural lineage, we used nuclear extracts of HSV infected nerve growth factor-differentiated PC12 and Vero cells, respectively, as the source of protein in gel shift assays. In both cell types, three complexes (complexes A, B, and C) which contain either OBP or OBPC were shown to bind specifically to a probe which contains the highest-affinity OBP binding site in oriS, site 1. Complex A was shown to contain OBPC exclusively, whereas complexes B and C contained OBP and likely other cellular proteins. By fine-mapping the binding sites of these three complexes, we identified single nucleotides which, when mutated, eliminated formation of all three complexes, or complexes B and C, but not A. In transient DNA replication assays, both mutations significantly impaired oriS-dependent DNA replication, demonstrating that formation of OBP containing complexes B and C is required for efficient initiation of oriS dependent DNA replication, whereas formation of the OBPC-containing complex A is insufficient for efficient initiation. PMID- 11435560 TI - The R region found in the human foamy virus long terminal repeat is critical for both Gag and Pol protein expression. AB - It has been suggested that sequences located within the 5' noncoding region of human foamy virus (HFV) are critical for expression of the viral Gag and Pol structural proteins. Here, we identify a discrete approximately 151-nucleotide sequence, located within the R region of the HFV long terminal repeat, that activates HFV Gag and Pol expression when present in the 5' noncoding region but that is inactive when inverted or when placed in the 3' noncoding region. Sequences that are critical for the expression of both Gag and Pol include not only the 5' splice site positioned at +51 in the R region, which is used to generate the spliced pol mRNA, but also intronic R sequences located well 3' to this splice site. Analysis of total cellular gag and pol mRNA expression demonstrates that deletion of the R region has little effect on gag mRNA levels but that R deletions that would be predicted to leave the pol 5' splice site intact nevertheless inhibit the production of the spliced pol mRNA. Gag expression can be largely rescued by the introduction of an intron into the 5' noncoding sequence in place of the R region but not by an intron or any one of several distinct retroviral nuclear RNA export sequences inserted into the mRNA 3' noncoding sequence. Neither the R element nor the introduced 5' intron markedly affects the cytoplasmic level of HFV gag mRNA. The poor translational utilization of these cytoplasmic mRNAs when the R region is not present in cis also extended to a cat indicator gene linked to an internal ribosome entry site introduced into the 3' noncoding region. Together these data imply that the HFV R region acts in the nucleus to modify the cytoplasmic fate of target HFV mRNA. The close similarity between the role of the HFV R region revealed in this study and previous data (M. Butsch, S. Hull, Y. Wang, T. M. Roberts, and K. Boris-Lawrie, J. Virol. 73:4847--4855, 1999) demonstrating a critical role for the R region in activating gene expression in the unrelated retrovirus spleen necrosis virus suggests that several distinct retrovirus families may utilize a common yet novel mechanism for the posttranscriptional activation of viral structural protein expression. PMID- 11435561 TI - Functional analysis of recombinant respiratory syncytial virus deletion mutants lacking the small hydrophobic and/or attachment glycoprotein gene. AB - Respiratory syncytial virus (RSV) produces three envelope glycoproteins, the attachment glycoprotein (G), the fusion (F) protein, and the small hydrophobic (SH) protein. It had been assumed, by analogy with other paramyxoviruses, that the G and F proteins would be required for the first two steps of viral entry, attachment and fusion. However, following repeated passage in cell culture, a viable mutant RSV that lacked both the G and SH genes was isolated (R. A. Karron, D. A. Buonagurio, A. F. Georgiu, S. S. Whitehead, J. E. Adamus, M. L. Clements Mann, D. O. Harris, V. B. Randolph, S. A. Udem, B. R. Murphy, and M. S. Sidhu, Proc. Natl. Acad. Sci. USA 94:13,961--13,966, 1997). To explore the roles of the G, F, and SH proteins in virion assembly, function, and cytopathology, we have modified the full-length RSV cDNA and used it to rescue infectious RSV lacking the G and/or SH genes. The three resulting viruses and the parental virus all contain the green fluorescent protein (GFP) gene that serves to identify infected cells. We have used purified, radiolabeled virions to examine virus production and function, in conjunction with GFP to quantify infected cells. We found that the G protein enhances virion binding to target cells but plays no role in penetration after attachment. The G protein also enhances cell-to-cell fusion, presumably via cell-to-cell binding, and enhances virion assembly or release. The presence or absence of the G protein in virions has no obvious effect on the content of F protein or host cell proteins in the virion. In growth curve experiments, the viruses lacking the G protein produced viral titers that were at least 10-fold lower than titers of viruses containing the G protein. This reduction is due in large part to the less efficient release of virions and the lower infectivity of the released virions. In the absence of the G protein, virus expressing both the F and SH proteins displayed somewhat smaller plaques, lower fusion activity, and slower viral entry than the virus expressing the F protein alone, suggesting that the SH protein has a negative effect on virus fusion in cell culture. PMID- 11435562 TI - Gag-Pol supplied in trans is efficiently packaged and supports viral function in human immunodeficiency virus type 1. AB - The intracellular trafficking and subsequent incorporation of Gag-Pol into human immunodeficiency virus type 1 (HIV-1) remains poorly defined. Gag-Pol is encoded by the same mRNA as Gag and is generated by ribosomal frameshifting. The multimerization of Gag and Gag-Pol is an essential step in the formation of infectious viral particles. In this study, we examined whether the interaction between Gag and Gag-Pol is initiated during protein translation in order to facilitate the trafficking and subsequent packaging of Gag-Pol into the virion. A conditional cotransfection system was developed in which virion formation required the coexpression of two HIV-1-based plasmids, one that produces both Gag and Gag-Pol and one that only produces Gag-Pol. The Gag-Pol proteins were either immunotagged with a His epitope or functionally tagged with a mutation (K65R) in reverse transcriptase that is associated with drug resistance. Gag-Pol packaging was assessed to determine whether the Gag-Pol incorporated into the virion was preferentially packaged from the plasmid that expressed both Gag and Gag-Pol or whether it could be packaged from either plasmid. Our data show that translation of Gag and Gag-Pol from the same mRNA is not critical for virion packaging of the Gag-Pol polyprotein or for viral function. PMID- 11435563 TI - Identification of envelope determinants of feline leukemia virus subgroup B that permit infection and gene transfer to cells expressing human Pit1 or Pit2. AB - The retroviral vector systems that are in common use for gene therapy are designed to infect cells expressing either of two widely expressed phosphate transporter proteins, Pit1 or Pit2. Subgroup B feline leukemia viruses (FeLV-Bs) use the gibbon ape leukemia virus receptor, Pit1, as a receptor for entry. Our previous studies showed that some chimeric envelope proteins encoding portions of FeLV-B could also enter cells by using a related receptor protein, Pit2, which serves as the amphotropic murine leukemia virus receptor (S. Boomer, M. Eiden, C. C. Burns, and J. Overbaugh, J. Virol. 71:8116--8123, 1997). Here we show that an arginine at position 73 within variable region A (VRA) of the FeLV-B envelope surface unit (SU) is necessary for viral entry into cells via the human Pit2 receptor. However, C-terminal SU sequences have a dominant effect in determining human Pit2 entry, even though this portion of the protein is outside known receptor binding domains. This suggests that a combination of specific VRA sequences and C-terminal sequences may influence interactions between FeLV-B SU and the human Pit2 receptor. Binding studies suggest that the C-terminal sequences may affect a postbinding step in viral entry via the Pit2 receptor, although in all cases, binding of FeLV-B SU to human Pit2 was weak. In contrast, neither the arginine 73 nor specific C-terminal sequences are required for efficient binding or infection with Pit1. Taken together, these data suggest that different residues in SU may interact with these two receptors. The specific FeLV Bs described here, which can enter cells using either human Pit receptor, may be useful as envelope pseudotypes for viruses used in gene therapy. PMID- 11435564 TI - Second-site suppressors of Rous sarcoma virus Ca mutations: evidence for interdomain interactions. AB - The capsid (CA) protein, the major structural component of retroviruses, forms a shell that encases the ribonucleoprotein complex in the virion core. The most conserved region of CA, approximately 20 amino acids of the major homology region (MHR), lies within the carboxy-terminal domain of the protein. Structural and sequence similarities among CA proteins of retroviruses and the CA-like proteins of hepatitis B virus and various retrotransposons suggest that the MHR is involved in an aspect of replication common to these reverse-transcribing elements. Conservative substitutions in this region of the Rous sarcoma virus protein were lethal due to a severe deficiency in reverse transcription, in spite of the presence of an intact genome and active reverse transcriptase in the particles. This finding suggests that the mutations interfered with normal interactions among these constituents. A total of four genetic suppressors of three lethal MHR mutations have now been identified. All four map to the sequence encoding the CA-spacer peptide (SP) region of Gag. The F167Y mutation in the MHR was fully suppressed by a single amino acid change in the alpha helix immediately downstream of the MHR, a region that forms the major dimer interface in human immunodeficiency virus CA. This finding suggests that the F167Y mutation indirectly interfered with dimerization. The F167Y defect could also be repaired by a second, independent suppressor in the C-terminal SP that was removed from CA during maturation. This single residue change, which increased the rate of SP cleavage, apparently corrected the F167Y defect by modifying the maturation pathway. More surprising was the isolation of suppressors of the R170Q and L171V MHR mutations, which mapped to the N-terminal domain of the CA protein. This finding suggests that the two domains, which in the monomeric protein are separated by a flexible linker, must communicate with each other at some unidentified point in the viral replication cycle. PMID- 11435565 TI - Identification of a conserved residue of foamy virus Gag required for intracellular capsid assembly. AB - In contrast to all retroviruses but similar to the hepatitis B virus, foamy viruses (FV) require expression of the envelope protein for budding of intracellular capsids from the cell, suggesting a specific interaction between the Gag and Env proteins. Capsid assembly occurs in the cytoplasm of infected cells in a manner similar to that for the B- and D-type viruses; however, in contrast to these retroviruses, FV Gag lacks an N-terminal myristylation signal and capsids are not targeted to the plasma membrane (PM). We have found that mutation of an absolutely conserved arginine (Arg) residue at position 50 to alanine (R50A) of the simian foamy virus SFV cpz(hu) inhibits proper capsid assembly and abolishes viral budding even in the presence of the envelope (Env) glycoproteins. Particle assembly and extracellular release of virus can be restored to this mutant with the addition of an N-terminal Src myristylation signal (Myr-R50A), presumably by providing an alternate site for assembly to occur at the PM. In addition, the strict requirement of Env expression for capsid budding can be bypassed by addition of a PM-targeting signal to Gag. These results suggest that intracellular capsid assembly may be mediated by a signal akin to the cytoplasmic targeting and retention signal CTRS found in Mason-Pfizer monkey virus and that FV Gag has the inherent ability to assemble capsids at multiple sites like conventional retroviruses. The necessity of Env expression for particle egress is most probably due to the lack of a membrane-targeting signal within FV Gag to direct capsids to the PM for release and indicates that Gag-Env interactions are essential to drive particle budding. PMID- 11435566 TI - Cytomegalovirus basic phosphoprotein (pUL32) binds to capsids in vitro through its amino one-third. AB - The cytomegalovirus (CMV) basic phosphoprotein (BPP) is a component of the tegument. It remains with the nucleocapsid fraction under conditions that remove most other tegument proteins from the virion, suggesting a direct and perhaps tight interaction with the capsid. As a step toward localizing this protein within the molecular structure of the virion and understanding its function during infection, we have investigated the BPP-capsid interaction. In this report we present evidence that the BPP interacts selectively, through its amino one third, with CMV capsids. Radiolabeled simian CMV (SCMV) BPP, synthesized in vitro, bound to SCMV B-capsids, and C-capsids to a lesser extent, following incubation with either isolated capsids or lysates of infected cells. Human CMV (HCMV) BPP (pUL32) also bound to SCMV capsids, and SCMV BPP likewise bound to HCMV capsids, indicating that the sequence(s) involved is conserved between the two proteins. Analysis of SCMV BPP truncation mutants localized the capsid binding region to the amino one-third of the molecule--the portion of BPP showing the greatest sequence conservation between the SCMV and HCMV homologs. This general approach may have utility in studying the interactions of other proteins with conformation-dependent binding sites. PMID- 11435567 TI - Characterization of nuclear RNases that cleave hepatitis B virus RNA near the La protein binding site. AB - Hepatitis B virus (HBV) RNA is downregulated by inflammatory cytokines induced in the liver by adoptively transferred HBV-specific cytotoxic T lymphocytes (CTLs) and during murine cytomegalovirus (MCMV) infections of the livers of HBV transgenic mice. The disappearance of HBV RNA is tightly associated with the cytokine-induced proteolytic cleavage of a previously defined HBV RNA-binding protein known as La autoantigen. La binds to a predicted stem-loop structure at the 5' end of the posttranscriptional regulatory element of HBV RNA between nucleotides 1243 and 1333. In the present study, we searched for nuclear RNase activities that might be involved in HBV RNA decay. Nuclear extracts derived from control livers and CTL-injected and MCMV-infected livers were analyzed for the ability to cleave HBV RNA. Endonucleolytic activity that cleaved HBV RNA at positions 1269 to 1270 and 1271 to 1272, immediately 5' of the stem-loop bound by the La protein (positions 1272 to 1293), was detected. Furthermore, we provide evidence that the cytokine-dependent downregulation of HBV RNA following MCMV infection is temporally associated with the upregulation of the endonucleolytic activity herein described. Collectively, these results suggest a model in which the steady-state HBV RNA content is controlled by the stabilizing influence of La and the destabilizing influence of nuclear RNase activities. PMID- 11435568 TI - Adeno-associated virus serotype 4 (AAV4) and AAV5 both require sialic acid binding for hemagglutination and efficient transduction but differ in sialic acid linkage specificity. AB - Adeno-associated virus serotype 4 (AAV4) and AAV5 have different tropisms compared to AAV2 and to each other. We recently reported that alpha 2--3 sialic acid is required for AAV5 binding and transduction. In this study, we characterized AAV4 binding and transduction and found it also binds sialic acid, but the specificity is significantly different from AAV5. AAV4 can hemagglutinate red blood cells from several species, whereas AAV5 hemagglutinates only rhesus monkey red blood cells. Treatment of red blood cells with trypsin inhibited hemagglutination for both AAV4 and AAV5, suggesting that the agglutinin is a protein. Treatment of Cos and red blood cells with neuraminidases also indicated that AAV4 bound alpha 2--3 sialic acid. However, resialylation experiments with neuraminidase-treated red blood cells demonstrated that AAV4 binding required alpha 2--3 O-linked sialic acid, whereas AAV5 required N-linked sialic acid. Similarly, resialylation of sialic acid-deficient CHO cells supported this same conclusion. The difference in linkage specificity for AAV4 and AAV5 was confirmed by binding and transduction experiments with cells incubated with either N-linked or O-linked inhibitors of glycosylation. Furthermore, AAV4 transduction was only blocked with soluble alpha 2-3 sialic acid, whereas AAV5 could be blocked with either alpha 2--3 or alpha 2-6 sialic acid. These results suggest that AAV4 and AAV5 require different sialic acid-containing glycoproteins for binding and transduction of target cells and they further explain the different tropism of AAV4 and AAV5. PMID- 11435569 TI - Octamer-binding sequence is a key element for the autoregulation of Kaposi's sarcoma-associated herpesvirus ORF50/Lyta gene expression. AB - The expression of the Kaposi's sarcoma-associated herpesvirus (KSHV) open reading frame 50 (ORF50) protein, Lyta (lytic transactivator), marks the switch from latent KSHV infection to the lytic phase. ORF50/Lyta upregulates several target KSHV genes, such as K8 (K-bZip), K9 (vIRF1), and ORF57, finally leading to the production of mature viruses. The auto-upregulation of ORF50/Lyta is thought to be an important mechanism for efficient lytic viral replication. In this study, we focused on this autoregulation and identified the promoter element required for it. An electrophoretic mobility shift assay indicated that the octamer binding protein 1 (Oct-1) bound to this element. Mutations in the octamer-binding motif resulted in refractoriness of the ORF50/Lyta promoter to transactivation by ORF50/Lyta, and Oct-1 expression enhanced this transactivation. These results suggest that the autoregulation of ORF50/Lyta is mediated by Oct-1. PMID- 11435570 TI - Polymerase slippage at vesicular stomatitis virus gene junctions to generate poly(A) is regulated by the upstream 3'-AUAC-5' tetranucleotide: implications for the mechanism of transcription termination. AB - Termination of mRNA synthesis in vesicular stomatitis virus (VSV), the prototypic rhabdovirus, is controlled by a 13-nucleotide gene end sequence which comprises the conserved tetranucleotide 3'-AUAC-5', the U(7) tract and the intergenic dinucleotide. mRNAs terminated at this sequence possess 100- to 300-nucleotide long 3' poly(A) tails which are thought to result from polymerase slippage (reiterative transcription) by the VSV polymerase on the U(7) tract. Previously we determined that in addition to the AUAC tetranucleotide, the U(7) tract was an essential signal in the termination process. Shortening or interrupting the U(7) tract abolished termination. These altered U tracts also prevented the polymerase from performing reiterative transcription necessary for generation of the mRNA poly(A) tail and thus established seven residues as the minimum length of U tract that allowed reiterative transcription to occur. In this study we investigated whether sequences other than the essential U(7) tract are involved in controlling polymerase slippage. We investigated whether the AUAC tetranucleotide affected the process of reiterative transcription by analyzing the nucleotide sequence of RNAs transcribed from altered subgenomic templates and infectious VSV variants. The tetranucleotide was found to regulate reiterative transcription on the U(7) tract. The extent of polymerase slippage was governed not by specific tetranucleotide sequences but rather by nucleotide composition such that slippage occurred when the tetranucleotide was composed of A or U residues but not when it was composed of G or C residues. This suggested that polymerase slippage was controlled, at least in part, by the strength of base pairing between the template and nascent strands. Further data presented here indicate that the tetranucleotide contains both a signal that directs the VSV polymerase to slip on the downstream U(7) tract and also a signal that directs a slipping polymerase to terminate mRNA synthesis. PMID- 11435571 TI - Characterization of a late entry event in the replication cycle of human immunodeficiency virus type 2. AB - Certain human cell lines and primary macrophage cultures are restricted to infection by some primary isolates of human immunodeficiency virus type 2 (HIV 2), although early steps of the viral life cycle such as fusion at the plasma membrane and reverse transcription are fully supported. The late postintegration events, transcription, translation, assembly, budding, and maturation into infectious virions are functional in restrictive cells. Apart from primary macrophages, the restrictive cell types are actively dividing, and nuclear import of preintegration complexes (PICs) is not required for infection. We therefore postulate that the PICs are trapped in a cellular compartment, preventing subsequent steps in the replication cycle that lead to integration of the provirus. To test this we showed that HIV-2 particles pseudotyped with vesicular stomatitis virus envelope G protein, which delivers HIV into an endocytic compartment, could overcome the block to infection. We suggest that delivery of the viral core into an appropriate cellular compartment is a critical step during the entry process of HIV. PMID- 11435572 TI - Effects of DNA structure and homology length on vaccinia virus recombination. AB - Replicating poxviruses catalyze high-frequency recombination reactions by a process that is not well understood. Using transfected DNA substrates we show that these viruses probably use a single-strand annealing recombination mechanism. Plasmids carrying overlapping portions of a luciferase gene expression cassette and luciferase assays were first shown to provide an accurate method of assaying recombinant frequencies. We then transfected pairs of DNAs into virus infected cells and monitored the efficiencies of linear-by-linear, linear-by circle, and circle-by-circle recombination. These experiments showed that vaccinia virus recombination systems preferentially catalyze linear-by-linear reactions much more efficiently than circle-by-circle reactions and catalyze circle-by-circle reactions more efficiently than linear-by-circle reactions. Reactions involving linear substrates required surprisingly little sequence identity, with only 16-bp overlaps still permitting approximately 4% recombinant production. Masking the homologies by adding unrelated DNA sequences to the ends of linear substrates inhibited recombination in a manner dependent upon the number of added sequences. Circular molecules were also recombined by replicating viruses but at frequencies 15- to 50-fold lower than are linear substrates. These results are consistent with mechanisms in which exonuclease or helicase processing of DNA ends permits the forming of recombinants through annealing of complementary single strands. Our data are not consistent with a model involving strand invasion reactions, because such reactions should favor mixtures of linear and circular substrates. We also noted that many of the reaction features seen in vivo were reproduced in a simple in vitro reaction requiring only purified vaccinia virus DNA polymerase, single-strand DNA binding protein, and pairs of linear substrates. The 3'-to-5' exonuclease activity of poxviral DNA polymerases potentially catalyzes recombination in vivo. PMID- 11435573 TI - The number of a U3 repeat box acting as an enhancer in long terminal repeats of polytropic replication-competent porcine endogenous retroviruses dynamically fluctuates during serial virus passages in human cells. AB - The organization and transcriptional regulation of porcine endogenous retrovirus (PERV) long terminal repeats (LTRs) are unknown. We have studied the activity of LTRs from replication-competent molecular clones by performing luciferase reporter assays. The LTRs differ in the presence and number of 39-bp repeats located in U3 that confer strong promoter activity in human, simian, canine, feline, and porcine cell lines, whereas for LTRs devoid of the repeats, the promoter strength was significantly reduced. As the activity of a heterologous simian virus 40 promoter and a homologous repeat-deficient LTR was elevated by four 39-bp repeats independently of its orientation and location, the repeat box complies with the definition of an enhancer. During serial virus passaging of molecular PERV clones on human 293 cells, proviral LTRs demonstrated adaptation of transcriptional activity by dynamic changes of the number of 39-bp repeats in the course of up to 12 passaging cycles. PMID- 11435574 TI - Selective regulation of human immunodeficiency virus-infected CD4(+) lymphocytes by a synthetic immunomodulator leads to potent virus suppression in vitro and in hu-PBL-SCID mice. AB - We have previously observed that the synthetic immunomodulator Murabutide inhibits human immunodeficiency virus type 1 (HIV-1) replication at multiple levels in macrophages and dendritic cells. The present study was designed to profile the activity of Murabutide on CD8-depleted phytohemagglutinin-activated lymphocytes from HIV-1-infected subjects and on the outcome of HIV-1 infection in severe combined immunodeficiency mice reconstituted with human peripheral blood leukocytes (hu-PBL-SCID mice). Maintaining cultures of CD8-depleted blasts from 36 patients in the presence of Murabutide produced dramatically reduced levels of viral p24 protein in the supernatants. This activity correlated with reduced viral transcripts and proviral DNA, was evident in cultures harboring R5, X4-R5, or X4 HIV-1 isolates, was not linked to inhibition of cellular DNA synthesis, and did not correlate with beta-chemokine release. Moreover, c-myc mRNA expression was down-regulated in Murabutide-treated cells, suggesting potential interference of the immunomodulator with the nuclear transport of viral preintegration complexes. On the other hand, daily treatment of HIV-1-infected hu-PBL-SCID mice with Murabutide significantly reduced the viral loads in plasma and the proviral DNA content in human peritoneal cells. These results are the first to demonstrate that a clinically acceptable synthetic immunomodulator with an ability to enhance the host's nonspecific immune defense mechanisms against infections can directly regulate cellular factors in infected lymphocytes, leading to controlled HIV-1 replication. PMID- 11435575 TI - Antibody from patients with acute human immunodeficiency virus (HIV) infection inhibits primary strains of HIV type 1 in the presence of natural-killer effector cells. AB - The partial control of viremia during acute human immunodeficiency virus type 1 (HIV-1) infection is accompanied by an HIV-1-specific cytotoxic T-lymphocyte (CTL) response and an absent or infrequent neutralizing antibody response. The control of HIV-1 viremia has thus been attributed primarily, if not exclusively, to CTL activity. In this study, the role of antibody in controlling viremia was investigated by measuring the ability of plasma or immunoglobulin G from acutely infected patients to inhibit primary strains of HIV-1 in the presence of natural killer (NK) effector cells. Antibody that inhibits virus when combined with effector cells was present in the majority of patients within days or weeks after onset of symptoms of acute infection. Furthermore, the magnitude of this effector cell-mediated antiviral antibody response was inversely associated with plasma viremia level, and both autologous and heterologous HIV-1 strains were inhibited. Finally, antibody from acutely infected patients likely reduced HIV-1 yield in vitro both by mediating effector cell lysis of target cells expressing HIV-1 glycoproteins and by augmenting the release of beta-chemokines from NK cells. HIV 1-specific antibody may be an important contributor to the early control of HIV viremia. PMID- 11435576 TI - Human hepatitis B virus polymerase interacts with the molecular chaperonin Hsp60. AB - Previous studies showed that hepatitis B virus polymerase (HBV Pol) interacts with host factors such as the Hsp90 complex, which is a critical step in viral genome replication. In this report, we propose that another chaperone, Hsp60, interacts with human HBV Pol and that this is a very important step for maturation of human HBV Pol into the active state. In the immunoprecipitation of recombinant human HBV Pol expressed in insect cells with the recombinant baculovirus expression system, the 60-kDa protein was coimmunoprecipitated with Pol and the protein was identified as Hsp60 through peptide sequencing and immunogenic analysis with an anti-Hsp60 antibody. In vitro experiments showed that Hsp60 strongly affected human HBV Pol activity in that (i) blocking of Hsp60 by the protein-specific antibody reduced human HBV Pol activity, (ii) the activity was increased by addition of Hsp60 in the presence of ATP, and (iii) ATP synergistically activated human HBV Pol with Hsp60. In vivo experiments showed that inhibition of Hsp60 in cells by a mutant Hsp60, C Delta 540, resulted in the reduction of human HBV Pol activity. In summary, our results indicate that the interaction is significant for conversion of human HBV Pol into the active state. PMID- 11435577 TI - Extrachromosomal recombinant adeno-associated virus vector genomes are primarily responsible for stable liver transduction in vivo. AB - Recombinant adeno-associated virus (rAAV) vectors stably transduce hepatocytes in experimental animals. Although the vector genomes are found both as extrachromosomes and as chromosomally integrated forms in hepatocytes, the relative proportion of each has not yet been clearly established. Using an in vivo assay based on the induction of hepatocellular regeneration via a surgical two-thirds partial hepatectomy, we have determined the proportion of integrated and extrachromosomal rAAV genomes in mouse livers and their relative contribution to stable gene expression in vivo. Plasma human coagulation factor IX (hF.IX) levels in mice originating from a chromosomally integrated hF.IX-expressing transposon vector remained unchanged with hepatectomy. This was in sharp contrast to what was observed when a surgical partial hepatectomy was performed in mice 6 weeks to 12 months after portal vein injection of a series of hF.IX-expressing rAAV vectors. At doses of 2.4 x 10(11) to 3.0 x 10(11) vector genomes per mouse (n = 12), hF.IX levels and the average number of stably transduced vector genomes per cell decreased by 92 and 86%, respectively, after hepatectomy. In a separate study, one of three mice injected with a higher dose of rAAV had a higher proportion (67%) of integrated genomes, the significance of which is not known. Nevertheless, in general, these results indicate that, in most cases, no more than approximately 10% of stably transduced genomes integrated into host chromosomes in vivo. Additionally, the results demonstrate that extrachromosomal, not integrated, genomes are the major form of rAAV in the liver and are the primary source of rAAV-mediated gene expression. This small fraction of integrated genomes greatly decreases the potential risk of vector-related insertional mutagenesis associated with all integrating vectors but also raises uncertainties as to whether rAAV-mediated hepatic gene expression can persist lifelong after a single vector administration. PMID- 11435578 TI - Suboptimal enhancer sequences are required for efficient bovine leukemia virus propagation in vivo: implications for viral latency. AB - Repression of viral expression is a major strategy developed by retroviruses to escape from the host immune response. The absence of viral proteins (or derived peptides) at the surface of an infected cell does not permit the establishment of an efficient immune attack. Such a strategy appears to have been adopted by animal oncoviruses such as bovine leukemia virus (BLV) and human T-cell leukemia virus (HTLV). In BLV-infected animals, only a small fraction of the infected lymphocytes (between 1 in 5,000 and 1 in 50,000) express large amounts of viral proteins; the vast majority of the proviruses are repressed at the transcriptional level. Induction of BLV transcription involves the interaction of the virus-encoded Tax protein with the CREB/ATF factors; the resulting complex is able to interact with three 21-bp Tax-responsive elements (TxRE) located in the 5' long terminal repeat (5' LTR). These TxRE contain cyclic AMP-responsive elements (CRE), but, remarkably, the "TGACGTCA" consensus is never strictly conserved in any viral strain (e.g.,AGACGTCA, TGACGGCA, TGACCTCA). To assess the role of these suboptimal CREs, we introduced a perfect consensus sequence within the TxRE and showed by gel retardation assays that the binding efficiency of the CREB/ATF proteins was increased. However, trans-activation of a luciferase-based reporter by Tax was not affected in transient transfection assays. Still, in the absence of Tax, the basal promoter activity of the mutated LTR was increased as much as 20-fold. In contrast, mutation of other regulatory elements within the LTR (the E box, NF-kappa B, and glucocorticoid- or interferon-responsive sites [GRE or IRF]) did not induce a similar alteration of the basal transcription levels. To evaluate the biological relevance of these observations made in vitro, the mutations were introduced into an infectious BLV molecular clone. After injection into sheep, it appeared that all the recombinants were infectious in vivo and did not revert into a wild-type virus. All of them, except one, propagated at wild-type levels, indicating that viral spread was not affected by the mutation. The sole exception was the CRE mutant; proviral loads were drastically reduced in sheep infected with this type of virus. We conclude that a series of sites (NF-kappa B, IRF, GRE, and the E box) are not required for efficient viral spread in the sheep model, although mutation of some of these motifs might induce a minor phenotype during transient transfection assays in vitro. Remarkably, a provirus (pBLV-Delta 21-bp) harboring only two TxRE was infectious and propagated at wild-type levels. And, most importantly, reconstitution of a consensus CRE, within the 21-bp enhancers increases binding of CREB/ATF proteins but abrogates basal repression of LTR-directed transcription in vitro. Suboptimal CREs are, however, essential for efficient viral spread within infected sheep, although these sites are dispensable for infectivity. These results suggest an evolutionary selection of suboptimal CREs that repress viral expression with escape from the host immune response. These observations, which were obtained in an animal model for HTLV-1, are of interest for oncovirus induced pathogenesis in humans. PMID- 11435579 TI - Genomic stability of murine leukemia viruses containing insertions at the Env-3' untranslated region boundary. AB - Retroviruses containing inserts of exogenous sequences frequently eliminate the inserted sequences upon spread in susceptible cells. We have constructed replication-competent murine leukemia virus (MLV) vectors containing internal ribosome entry site (IRES)-transgene cassettes at the env-3' untranslated region boundary in order to examine the effects of insert sequence and size on the loss of inserts during viral replication. A virus containing an insertion of 1.6 kb replicated with greatly attenuated kinetics relative to wild-type virus and lost the inserted sequences in a single infection cycle. In contrast, MLVs containing inserts of 1.15 to 1.30 kb replicated with kinetics only slightly attenuated compared to wild-type MLV and exhibited much greater stability, maintaining their genomic integrity over multiple serial infection cycles. Eventually, multiple species of deletion mutants were detected simultaneously in later infection cycles; once detected, these variants rapidly dominated the population and thereafter appeared to be maintained at a relative equilibrium. Sequence analysis of these variants identified preferred sites of recombination in the parental viruses, including both short direct repeats and inverted repeats. One instance of insert deletion through recombination with an endogenous retrovirus was also observed. When specific sequences involved in these recombination events were eliminated, deletion variants still arose with the same kinetics upon virus passage and by apparently similar mechanisms, although at different locations in the vectors. Our results suggest that while lengthened, insert-containing genomes can be maintained over multiple replication cycles, preferential deletions resulting in loss of the inserted sequences confer a strong selective advantage. PMID- 11435580 TI - Phylogenetic and evolutionary relationships among yellow fever virus isolates in Africa. AB - Previous studies with a limited number of strains have indicated that there are two genotypes of yellow fever (YF) virus in Africa, one in west Africa and the other in east and central Africa. We have examined the prM/M and a portion of the E protein for a panel of 38 wild strains of YF virus from Africa representing different countries and times of isolation. Examination of the strains revealed a more complex genetic relationship than previously reported. Overall, nucleotide substitutions varied from 0 to 25.8% and amino acid substitutions varied from 0 to 9.1%. Phylogenetic analysis using parsimony and neighbor-joining algorithms identified five distinct genotypes: central/east Africa, east Africa, Angola, west Africa I, and west Africa II. Extensive variation within genotypes was observed. Members of west African genotype II and central/east African genotype differed by 2.8% or less, while west Africa genotype I varied up to 6.8% at the nucleotide level. We speculate that the former two genotypes exist in enzootic transmission cycles, while the latter is genetically more heterogeneous due to regular human epidemics. The nucleotide sequence of the Angola genotype diverged from the others by 15.7 to 23.0% but only 0.4 to 5.6% at the amino acid level, suggesting that this genotype most likely diverged from a progenitor YF virus in east/central Africa many years ago, prior to the separation of the other east/central African strains analyzed in this study, and has evolved independently. These data demonstrate that there are multiple genotypes of YF virus in Africa and suggest independent evolution of YF virus in different areas of Africa. PMID- 11435581 TI - Minute virus of mice initiator protein NS1 and a host KDWK family transcription factor must form a precise ternary complex with origin DNA for nicking to occur. AB - Parvoviral rolling hairpin replication generates palindromic genomic concatemers whose junctions are resolved to give unit-length genomes by a process involving DNA replication initiated at origins derived from each viral telomere. The left end origin of minute virus of mice (MVM), oriL, contains binding sites for the viral initiator nickase, NS1, and parvovirus initiation factor (PIF), a member of the emerging KDWK family of transcription factors. oriL is generated as an active form, oriL(TC), and as an inactive form, oriL(GAA), which contains a single additional nucleotide inserted between the NS1 and PIF sites. Here we examined the interactions on oriL(TC) which lead to activation of NS1 by PIF. The two subunits of PIF, p79 and p96, cooperatively bind two ACGT half-sites, which can be flexibly spaced. When coexpressed from recombinant baculoviruses, the PIF subunits preferentially form heterodimers which, in the presence of ATP, show cooperative binding with NS1 on oriL, but this interaction is preferentially enhanced on oriL(TC) compared to oriL(GAA). Without ATP, NS1 is unable to bind stably to its cognate site, but PIF facilitates this interaction, rendering the NS1 binding site, but not the nick site, resistant to DNase I. Varying the spacing of the PIF half-sites shows that the distance between the NS1 binding site and the NS1-proximal half-site is critical for nickase activation, whereas the position of the distal half-site is unimportant. When expressed separately, both PIF subunits form homodimers that bind site specifically to oriL, but only complexes containing p79 activate the NS1 nickase function. PMID- 11435582 TI - The vaccinia virus superoxide dismutase-like protein (A45R) is a virion component that is nonessential for virus replication. AB - A characterization of the A45R gene from vaccinia virus (VV) strain Western Reserve is presented. The open reading frame is predicted to encode a 125-amino acid protein (M(r), of 13,600) with 39% amino acid identity to copper-zinc superoxide dismutase (Cu-Zn SOD). Sequencing of the A45R gene from other orthopoxviruses, here and by others, showed that the protein is highly conserved in all viruses sequenced, including 16 strains of VV, 2 strains of cowpox virus, camelpox virus, and 4 strains of variola virus. In all cases the protein lacks key residues involved in metal ion binding that are important for the catalytic activity. The A45R protein was expressed in Escherichia coli, purified, and tested for SOD activity, but neither enzymatic nor inhibitory SOD activity was detected. Additionally, no virus-encoded SOD activity was detected in infected cells or purified virions. A monoclonal antibody raised against the A45R protein expressed in E. coli identified the A45R gene product as a 13.5-kDa protein that is expressed late during VV infection. Confocal microscopy of VV-infected cells indicated that the A45R protein accumulated predominantly in cytoplasmic viral factories. Electron microscopy and biochemical analyses showed that the A45R protein is incorporated into the virion core. A deletion mutant lacking the majority of the A45R gene and a revertant virus in which the deleted gene was restored were constructed and characterized. The growth properties of the deletion mutant virus were indistinguishable from those of wild-type and revertant viruses in all cell lines tested, including macrophages. Additionally, the virulence and pathogenicity of the three viruses were also comparable in murine and rabbit models of infection. A45R is unusual in being the first VV core protein described that affects neither virus replication nor virulence. PMID- 11435583 TI - Therapeutic effect of a Gag-nuclease fusion protein against retroviral infection in vivo. AB - Recently, remarkable progress has been made in developing effective combination drug therapies that can control but not cure retroviral replication. Even when effective, these drug regimens are toxic, they require demanding administration schedules, and resistant viruses can emerge. Thus the need for new gene-based therapies continues. In one such approach, capsid-targeted viral inactivation (CTVI), nucleases fused to viral coat proteins are expressed in infected cells and become incorporated during virion assembly. CTVI can eliminate infectious murine retrovirus titer in tissue culture. Here we describe transgenic mice expressing fusions of the Moloney murine leukemia virus (Mo-MuLV) Gag protein to staphylococcal nuclease. This work tests the protective effect and demonstrates in vivo proof-of-principle of CTVI in transgenic mice expressing endogenous proviral copies of Mo-MuLV. The antiviral protein-expressing mice are phenotypically normal, attesting to the lack of toxicity of the fusion protein. The Mo-MuLV infection was much less virulent in transgenic littermates than in nontransgenic littermates. Gag-nuclease expression reduced infectious titers in blood up to 10-fold, decreased splenomegaly and leukemic infiltration, and increased life spans up to 2.5-fold in transgenic relative to nontransgenic infected animals. These results suggest that gene therapies based on similar fusion proteins, designed to attack human immunodeficiency virus or other retroviruses, could provide substantial therapeutic benefits. PMID- 11435584 TI - Hairpin loop structure in the 3' arm of the influenza A virus virion RNA promoter is required for endonuclease activity. AB - Previous studies have shown that the 5' arm of the influenza A virus virion RNA promoter requires a hairpin loop structure for efficient endonuclease activity of influenza virus RNA polymerase, an activity that is required for the cap snatching activity of primers from host pre-mRNA. Here we examine whether a hairpin loop is also required in the 3' arm of the viral RNA promoter. We study point mutations at each nucleotide position (1 to 12) within the 3' arm of the promoter as well as complementary "rescue" mutations which restored base pairing in the stem of a potential hairpin loop. Our results suggest that endonuclease activity is absolutely dependent on the presence of a 3' hairpin loop structure. This is the first direct evidence for RNA secondary structure within the 3' arm being required for a specific stage, i.e., endonuclease cleavage, in the influenza virus replicative cycle. PMID- 11435585 TI - Functional interaction between fluorodeoxyuridine-induced cellular alterations and replication of a ribonucleotide reductase-negative herpes simplex virus. AB - G207 is an oncolytic herpes simplex virus (HSV) which is attenuated by inactivation of viral ribonucleotide reductase (RR) and deletion of both gamma(1)34.5 genes. The cellular counterparts that can functionally substitute for viral RR and the carboxyl-terminal domain of ICP34.5 are cellular RR and the corresponding homologous domain of the growth arrest and DNA damage protein 34 (GADD34), respectively. Because the thymidylate synthetase (TS) inhibitor fluorodeoxyuridine (FUdR) can alter expression of cellular RR and GADD34, we examined the effect of FUdR on G207 bioactivity with the hypothesis that FUdR induced cellular changes will alter viral proliferation and cytotoxicity. Replication of wild-type HSV-1 was impaired in the presence of 10 nM FUdR, whereas G207 demonstrated increased replication under the same conditions. Combined use of FUdR and G207 resulted in synergistic cytotoxicity. FUdR exposure caused elevation of RR activity at 10 and 100 nM, whereas GADD34 was induced only at 100 nM. The effect of enhanced viral replication by FUdR was suppressed by hydroxyurea, a known inhibitor of RR. These results demonstrate that the growth advantage of G207 in FUdR-treated cells is primarily based on an RR-dependent mechanism. Although our findings show that TS inhibition impairs viral replication, the FUdR-induced RR elevation may overcome this disadvantage, resulting in enhanced replication of G207. These data provide the cellular basis for the combined use of RR-negative HSV mutants and TS inhibitors in the treatment of cancer. PMID- 11435586 TI - DNA microarray analysis of chimpanzee liver during acute resolving hepatitis C virus infection. AB - Hepatitis C virus (HCV) poses a worldwide health problem in that the majority of individuals exposed to HCV become chronically infected and are predisposed for developing significant liver disease. DNA microarray technology provides an opportunity to survey transcription modulation in the context of an infectious disease and is a particularly attractive approach in characterizing HCV-host interactions, since the mechanisms underlying viral persistence and disease progression are not understood and are difficult to study. Here, we describe the changes in liver gene expression during the course of an acute-resolving HCV infection in a chimpanzee. Clearance of viremia in this animal occurred between weeks 6 and 8, while clearance of residual infected hepatocytes did not occur until 14 weeks postinfection. The most notable changes in gene expression occurred in numerous interferon response genes (including all three classical interferon antiviral pathways) that increased dramatically, some as early as day 2 postinfection. The data suggest a biphasic mechanism of viral clearance dependent on both the innate and adaptive immune responses and provide insight into the response of the liver to a hepatotropic viral infection. PMID- 11435587 TI - Interferon-independent, human immunodeficiency virus type 1 gp120-mediated induction of CXCL10/IP-10 gene expression by astrocytes in vivo and in vitro. AB - The CXC chemokine gamma interferon (IFN-gamma)-inducible protein CXCL10/IP-10 is markedly elevated in cerebrospinal fluid and brain of individuals infected with human immunodeficiency virus type 1 (HIV-1) and is implicated in the pathogenesis of HIV-associated dementia (HAD). To explore the possible role of CXCL10/IP-10 in HAD, we examined the expression of this and other chemokines in the central nervous system (CNS) of transgenic mice with astrocyte-targeted expression of HIV gp120 under the control of the glial fibrillary acidic protein (GFAP) promoter, a murine model for HIV-1 encephalopathy. Compared with wild-type controls, CNS expression of the CC chemokine gene CCL2/MCP-1 and the CXC chemokine genes CXCL10/IP-10 and CXCL9/Mig was induced in the GFAP-HIV gp120 mice. CXCL10/IP-10 RNA expression was increased most and overlapped the expression of the transgene encoded HIV gp120 gene. Astrocytes and to a lesser extent microglia were identified as the major cellular sites for CXCL10/IP-10 gene expression. There was no detectable expression of any class of IFN or their responsive genes. In astrocyte cultures, soluble recombinant HIV gp120 protein was capable of directly inducing CXCL10/IP-10 gene expression a process that was independent of STAT1. These findings highlight a novel IFN- and STAT1-independent mechanism for the regulation of CXCL10/IP-10 expression and directly link expression of HIV gp120 to the induction of CXCL10/IP-10 that is found in HIV infection of the CNS. Finally, one function of IP-10 expression may be the recruitment of leukocytes to the CNS, since the brain of GFAP-HIV gp120 mice had increased numbers of CD3(+) T cells that were found in close proximity to sites of CXCL10/IP-10 RNA expression. PMID- 11435588 TI - N-terminal domain of Borna disease virus G (p56) protein is sufficient for virus receptor recognition and cell entry. AB - Borna disease virus (BDV) surface glycoprotein (GP) (p56) has a predicted molecular mass of 56 kDa. Due to extensive posttranslational glycosylation the protein migrates as a polypeptide of 84 kDa (gp84). The processing of gp84 by the cellular protease furin generates gp43, which corresponds to the C-terminal part of gp84. Both gp84 and gp43 have been implicated in viral entry involving receptor-mediated endocytosis and pH-dependent fusion. We have investigated the domains of BDV p56 involved in virus entry. For this, we used a pseudotype approach based on a recently developed recombinant vesicular stomatitis virus (VSV) in which the gene for green fluorescent protein was substituted for the VSV G protein gene (VSV Delta G*). Complementation of VSV Delta G* with BDV p56 resulted in infectious VSV Delta G* pseudotypes that contained both BDV gp84 and gp43. BDV-VSV chimeric GPs that contained the N-terminal 244 amino acids of BDV p56 and amino acids 421 to 511 of VSV G protein were efficiently incorporated into VSV Delta G* particles, and the resulting pseudotype virions were neutralized by BDV-specific antiserum. These findings indicate that the N terminal part of BDV p56 is sufficient for receptor recognition and virus entry. PMID- 11435589 TI - Wild Mandrillus sphinx are carriers of two types of lentivirus. AB - Mandrillus sphinx, a large primate living in Cameroon and Gabon and belonging to the Papionini tribe, was reported to be infected by a simian immunodeficiency virus (SIV) (SIVmndGB1) as early as 1988. Here, we have identified a second, highly divergent SIVmnd (designated SIVmnd-2). Genomic organization differs between the two viral types; SIVmnd-2 has the additional vpx gene, like other SIVs naturally infecting the Papionini tribe (SIVsm and SIVrcm) and in contrast to the other SIVmnd type (here designated SIVmnd-1), which is more closely related to SIVs infecting l'hoest (Cercopithecus lhoesti lhoesti) and sun-tailed (Cercopithecus lhoesti solatus) monkeys. Importantly, our epidemiological studies indicate a high prevalence of both types of SIVmnd; all 10 sexually mature wild living monkeys and 3 out of 17 wild-born juveniles tested were infected. The geographic distribution of SIVmnd seems to be distinct for the two types: SIVmnd 1 viruses were exclusively identified in mandrills from central and southern Gabon, whereas SIVmnd-2 viruses were identified in monkeys from northern and western Gabon, as well as in Cameroon. SIVmnd-2 full-length sequence analysis, together with analysis of partial sequences from SIVmnd-1 and SIVmnd-2 from wild born or wild-living mandrills, shows that the gag and pol regions of SIVmnd-2 are closest to those of SIVrcm, isolated from red-capped mangabeys (Cercocebus torquatus), while the env gene is closest to that of SIVmnd-1. pol and env sequence analyses of SIV from a related Papionini species, the drill (Mandrillus leucophaeus), shows a closer relationship of SIVdrl to SIVmnd-2 than to SIVmnd-1. Epidemiological surveys of human immunodeficiency virus revealed a case in Cameroon of a human infected by a virus serologically related to SIVmnd, raising the possibility that mandrills represent a viral reservoir for humans similar to sooty mangabeys in Western Africa and chimpanzees in Central Africa. PMID- 11435590 TI - Efficient lymphoreticular prion propagation requires PrP(c) in stromal and hematopoietic cells. AB - In most prion diseases, infectivity accumulates in lymphoreticular organs early after infection. Defects in hematopoietic compartments, such as impaired B-cell maturation, or in stromal compartments, such as abrogation of follicular dendritic cells, can delay or prevent lymphoreticular prion colonization. However, the nature of the compartment in which prion replication takes place is controversial, and it is unclear whether this compartment coincides with that expressing the normal prion protein (PrP(c)). Here we studied the distribution of infectivity in splenic fractions of wild-type and fetal liver chimeric mice carrying the gene that encodes PrP(c) (Prnp) solely on hematopoietic or on stromal cells. We fractionated spleens at various times after intraperitoneal challenge with prions and assayed infectivity by bioassay. Upon high-dose challenge, chimeras carrying PrP(c) on hematopoietic cells accumulated prions in stroma and in purified splenocytes. In contrast, after low-dose challenge ablation of Prnp in either compartment prevented splenic accumulation of infectivity, indicating that optimal prion replication requires PrP(c) expression by both stromal and hematopoietic compartments. PMID- 11435591 TI - Phage display of adenovirus type 5 fiber knob as a tool for specific ligand selection and validation. AB - Adenovirus (Ad) vectors are most potent for use as gene delivery vehicles to infect human cells in vitro and in vivo with high efficiency. The main limitation in utilization of Ad as a gene transfer vector is the lack of specificity. Genetic modifications of Ad capsid proteins resulting in incorporation of foreign polypeptide ligand sequences can redirect the vector towards target cells. However, in many cases the incorporated ligands lose specificity or lead to conformational changes influencing virion integrity. In order to select target specific ligands a priori structurally compatible with Ad, we propose a system for displaying polypeptide sequences in the context of the Ad fiber knob on the surfaces of filamentous bacteriophages. To establish this concept, we displayed the wild-type Ad serotype 5 knob and knobs containing c-Myc epitopes and six histidine sequences in the pJuFo phage system. The knobs remained trimeric and bound the coxsackievirus-Ad receptor, and the phage knob-displayed ligands recognized and bound their cognates in the phage-displayed knob context. Further development of this system may be useful for candidate ligand fidelity and Ad structural compatibility validation prior to Ad modification. PMID- 11435592 TI - Sindbis virus-induced neuronal death is both necrotic and apoptotic and is ameliorated by N-methyl-D-aspartate receptor antagonists. AB - Virus infection of neurons leads to different outcomes ranging from latent and noncytolytic infection to cell death. Viruses kill neurons directly by inducing either apoptosis or necrosis or indirectly as a result of the host immune response. Sindbis virus (SV) is an alphavirus that induces apoptotic cell death both in vitro and in vivo. However, apoptotic changes are not always evident in neurons induced to die by alphavirus infection. Time lapse imaging revealed that SV-infected primary cortical neurons exhibited both apoptotic and necrotic morphological features and that uninfected neurons in the cultures also died. Antagonists of the N-methyl-D-aspartate (NMDA) subtype of glutamate receptors protected neurons from SV-induced death without affecting virus replication or SV induced apoptotic cell death. These results provide evidence that SV infection activates neurotoxic pathways that result in aberrant NMDA receptor stimulation and damage to infected and uninfected neurons. PMID- 11435593 TI - Genome of lumpy skin disease virus. AB - Lumpy skin disease virus (LSDV), a member of the capripoxvirus genus of the Poxviridae, is the etiologic agent of an important disease of cattle in Africa. Here we report the genomic sequence of LSDV. The 151-kbp LSDV genome consists of a central coding region bounded by identical 2.4 kbp-inverted terminal repeats and contains 156 putative genes. Comparison of LSDV with chordopoxviruses of other genera reveals 146 conserved genes which encode proteins involved in transcription and mRNA biogenesis, nucleotide metabolism, DNA replication, protein processing, virion structure and assembly, and viral virulence and host range. In the central genomic region, LSDV genes share a high degree of colinearity and amino acid identity (average of 65%) with genes of other known mammalian poxviruses, particularly suipoxvirus, yatapoxvirus, and leporipoxviruses. In the terminal regions, colinearity is disrupted and poxvirus homologues are either absent or share a lower percentage of amino acid identity (average of 43%). Most of these differences involve genes and gene families with likely functions involving viral virulence and host range. Although LSDV resembles leporipoxviruses in gene content and organization, it also contains homologues of interleukin-10 (IL-10), IL-1 binding proteins, G protein-coupled CC chemokine receptor, and epidermal growth factor-like protein which are found in other poxvirus genera. These data show that although LSDV is closely related to other members of the Chordopoxvirinae, it contains a unique complement of genes responsible for viral host range and virulence. PMID- 11435594 TI - Functional analysis of adenovirus protein IX identifies domains involved in capsid stability, transcriptional activity, and nuclear reorganization. AB - The product of adenovirus (Ad) type 5 gene IX (pIX) is known to actively participate in the stability of the viral icosahedron, acting as a capsid cement. We have previously demonstrated that pIX is also a transcriptional activator of several viral and cellular TATA-containing promoters, likely contributing to the transactivation of the Ad expression program. By extensive mutagenesis, we have now delineated the functional domains involved in each of the pIX properties: residues 22 to 26 of the highly conserved N-terminal domain are crucial for incorporation of the protein into the virion; specific residues of the C-terminal leucine repeat are responsible for pIX interactions with itself and possibly other proteins, a property that is critical for pIX transcriptional activity. We also show that pIX takes part in the virus-induced nuclear reorganization of late infected cells: the protein induces, most likely through self-assembly, the formation of specific nuclear structures which appear as dispersed nuclear globules by immunofluorescence staining and as clear amorphous spherical inclusions by electron microscopy. The integrity of the leucine repeat appears to be essential for the formation and nuclear retention of these inclusions. Together, our results demonstrate the multifunctional nature of pIX and provide new insights into Ad biology. PMID- 11435595 TI - Intrahepatic genetic inoculation of hepatitis C virus RNA confers cross protective immunity. AB - Naturally occurring hepatitis C virus (HCV) infection has long been thought to induce a weak immunity which is insufficient to protect an individual from subsequent infections and has cast doubt on the ability to develop effective vaccines. A series of intrahepatic genetic inoculations (IHGI) with type 1a HCV RNA were performed in a chimpanzee to determine whether a form of genetic immunization might stimulate protective immunity. We demonstrate that the chimpanzee not only developed protective immunity to the homologous type 1a RNA after rechallenge by IHGI but was also protected from chronic HCV infection after sequential rechallenge with 100 50% chimpanzee infectious doses of a heterologous type 1a (H77) and 1b (HC-J4) whole-virus inoculum. These results offer encouragement to pursue the development of HCV vaccines. PMID- 11435596 TI - Pocket protein p130/Rb2 is required for efficient herpes simplex virus type 1 gene expression and viral replication. AB - We have reported previously that herpes simplex virus type 1 (HSV-1) infection disrupts normal progression of the mammalian cell cycle, causing cells to enter a G(1)-like state. Infected cells were characterized by a decline in cyclin dependent kinase 2 (CDK2) activities, loss of hyperphosphorylated retinoblastoma protein (pRb), accumulation of E2F-pocket protein complexes, and failure to initiate cellular DNA replication. In the present study, we investigated the role of the pocket proteins pRb, p107, and p130 in HSV-1-dependent cell cycle inhibition and cyclin kinase regulation by infecting murine 3T3 cells derived from wild-type (WT) mouse embryos or embryos with deletions of pRb (pRb(-/-)), p107 (p107(-/-)), p130 (p130(-/-)), or both p130 and p107 (p130(-/-)/p107(-/-)). With respect to CDK2 inhibition, viral protein accumulation, viral DNA replication, and progeny virus yield, WT, pRb(-/-), and p107(-/-) cells were essentially identical. In contrast, after infection of p130(-/-) cells, we observed no inhibition of CDK2 activity, a 5- to 6-h delay in accumulation of viral proteins, an impaired ability to form viral DNA replication compartments, and reduced viral DNA synthesis. As a result, progeny virus yield was reduced 2 logs compared to that in WT cells. Notably, p130(-/-)/p107(-/-) double-knockout cells had a virus replication phenotype intermediate between those of the p107(-/ ) and p130(-/-) cells. We conclude from these studies that p130 is a key factor in regulating aspects of cell cycle progression, as well as the timely expression of viral genes and replication of viral DNA. PMID- 11435597 TI - Transcription pattern of human herpesvirus 8 open reading frame K3 in primary effusion lymphoma and Kaposi's sarcoma. AB - Human herpesvirus 8 (HHV-8) is found in immunoblastic B cells of patients with multicentric Castleman's disease (MCD) and, predominantly in a latent form, in primary effusion lymphoma (PEL) cells and Kaposi's sarcoma (KS) spindle cells. Recent studies have shown that upon reactivation, HHV-8 expresses factors that downregulate major histocompatibility class I proteins and coactivation molecules and that may enable productively infected cells to escape cytotoxic T lymphocytes and natural killer cell responses. One of these viral factors is encoded by open reading frame (ORF) K3. Here we show that in PEL cells, ORF K3 is expressed through viral transcripts that are induced very early upon virus reactivation, including bicistronic RNA molecules containing coding sequences from viral ORFs K3 and 70. Specifically, we found that a bicistronic transcript was expressed in the absence of de novo protein synthesis, thereby identifying a novel HHV-8 immediate-early gene product. Several features of the RNA molecules encoding the K3 product, including multiple transcriptional start sites, multiple donor splicing sites, and potential alternative ATG usage, suggest that there exists a finely tuned modulation of ORF K3 expression. By contrast, ORF K3 transcripts are not detected in the majority of cells present in KS lesions that are latently infected by the virus, suggesting that there are other, as-yet-unknown mechanisms of immune evasion for infected KS spindle cells. Nevertheless, because HHV-8 viremia precedes the development of KS lesions and is associated with the recrudescence of MCD symptoms, the prompt expression of ORF K3 in productively infected circulating cells may be important for virus pathogenesis. Thus, molecules targeting host or viral factors that activate ORF K3 expression or inactivate the biological functions of the K3 product should be exploited for the prevention or treatment of HHV-8-associated diseases in at-risk individuals. PMID- 11435598 TI - Lipid rafts and pseudotyping. AB - Specific interactions between envelope and core proteins govern the membrane assembly of most enveloped viruses. Despite this, mixed infections lead to pseudotyping, the association of the viral cores of one virus with the envelopes of another. How does this occur? We show here that the detergent-insoluble lipid rafts of the plasma membrane function as a natural meeting point for the transmembrane and core components of a phylogenetically diverse collection of enveloped viruses. As a result, viral particles preferentially incorporate both the envelope components of other viruses as well as the extra- and intracellular constituents of host cell lipid rafts, including gangliosides, glycosyl phosphatidylinositol-anchored surface proteins, and intracellular signal transduction molecules. Pharmacological disruption of lipid rafts interferes with virus production. PMID- 11435599 TI - Efficacy of dideoxynucleosides against human foamy virus and relationship to its reverse transcriptase amino acid sequence and structure. AB - Human foamy virus (HFV), a retrovirus of simian origin which occasionally infects humans, is the basis of retroviral vectors in development for gene therapy. Clinical considerations of how to treat patients developing an uncontrolled infection by either HFV or HFV-based vectors need to be raised. We determined the susceptibility of the HFV to dideoxynucleosides and found that only zidovudine was equally efficient against the replication of human immunodeficiency virus type 1 (HIV-1) and HFV. By contrast, zalcitabine (ddC), lamivudine (3TC), stavudine (d4T), and didanosine (ddI) were 3-, 3-, 30-, and 46-fold less efficient against HFV than against HIV-1, respectively. Some amino acid residues known to be involved in HIV-1 resistance to ddC, 3TC, d4T, and ddI were found at homologous positions of HFV reverse transcriptase (RT). These critical amino acids are located at the same positions in the three-dimensional structure of HIV 1 and HFV RT, suggesting that both enzymes share common patterns of inhibition. PMID- 11435600 TI - Abundant early expression of gpUL4 from a human cytomegalovirus mutant lacking a repressive upstream open reading frame. AB - The human cytomegalovirus UL4 gene encodes a 48-kDa glycoprotein, expression of which is repressed at the translational level by a short upstream open reading frame (uORF2) within the UL4 transcript leader. Mutation of the uORF2 initiation codon in the viral genome eliminates ribosomal stalling at the uORF2 termination site, resulting in early and abundant gpUL4 protein synthesis. This mutation does not appear to affect viral replication kinetics in human fibroblasts. These results reveal that the unusual uORF2 inhibitory mechanism is a principal determinant of the abundance and timing of gpUL4 expression but is nonessential for replication in cell culture. PMID- 11435601 TI - Context-dependent phenotype of a human immunodeficiency virus type 1 nucleocapsid mutation. AB - The human immunodeficiency virus type 1 (HIV-1) nucleocapsid mutation R10A/K11A abolishes viral replication when present in proviral clone HIV-1(HXB-2), but it was found to have minimal effect on replication of the closely related HIV-1(NL4 3). Functional mapping demonstrated that a nonconservative amino acid change at nucleocapsid residue 24 (threonine in HIV-1(HXB-2), isoleucine in HIV-1(NL4-3)) is the major determinant of the different R10A/K11A phenotypes in these two proviruses. Threonine-isoleucine exchanges appear to modify the R10A/K11A phenotype via effects on virion RNA-packaging efficiency. The improved packaging seen with hydrophobic isoleucine is consistent with solution structures localizing this residue to a hydrophobic pocket that contacts guanosine bases in viral genomic RNA stem-loops critical for packaging. PMID- 11435602 TI - E box-dependent activation of telomerase by human papillomavirus type 16 E6 does not require induction of c-myc. AB - Human papillomavirus type 16 (HPV-16) E6 activates telomerase specifically in epithelial cells. The oncogene c-myc has also been shown to activate telomerase in several cell types. Here we show that while both HPV-16 E6 and c-myc require intact E boxes to transactivate the hTERT promoter, E6 does not induce hTERT transcription simply by inducing expression of c-myc. Moreover, hTERT transactivation by HPV-16 E6 correlates with its ability to bind the cellular E6 associated protein (E6AP), suggesting that E6 and E6AP may target a regulator of hTERT expression. PMID- 11435603 TI - Correlation between viral resistance to zidovudine and resistance at the reverse transcriptase level for a panel of human immunodeficiency virus type 1 mutants. AB - Using a large panel of human immunodeficiency virus type 1 site-directed mutants, we have observed a higher correlation than has previously been demonstrated between zidovudine (AZT)-triphosphate resistance data at the reverse transcriptase (RT) level and corresponding viral AZT resistance. This enhanced resistance effect at the RT level was seen with ATP and to a lesser extent with PP(i) when ATP was added at physiological concentrations. The ATP-dependent mechanism (analogous to pyrophosphorolysis) appears to be dominant in the mutants bearing the D67N and K70R or 69 insertion mutations, whereas the Q151M mutation seems independent of ATP for decreased binding to AZT-triphosphate. PMID- 11435604 TI - DNA and ATP binding activities of the baculovirus DNA helicase P143. AB - P143 is a DNA helicase that tightly binds both double-stranded and single stranded DNA. DNA-protein complexes rapidly dissociated in the presence of ATP and Mg(2+). This finding suggests that ATP hydrolysis causes a conformational change in P143 which decreases affinity for DNA. This supports the model of an inchworm mechanism of DNA unwinding. PMID- 11435605 TI - Adenovirus type 9 fiber knob binds to the coxsackie B virus-adenovirus receptor (CAR) with lower affinity than fiber knobs of other CAR-binding adenovirus serotypes. AB - The coxsackie B virus and adenovirus (Ad) receptor (CAR) functions as an attachment receptor for multiple Ad serotypes. Here we show that the Ad serotype 9 (Ad9) fiber knob binds to CAR with much reduced affinity compared to the binding by Ad5 and Ad12 fiber knobs as well as the knob of the long fiber of Ad41 (Ad41L). Substitution of Asp222 in Ad9 fiber knob with a lysine that is conserved in Ad5, Ad12, and Ad41L substantially improved Ad9 fiber knob binding to CAR, while the corresponding substitution in Ad5 (Lys442Asp) significantly reduced Ad5 binding. The presence of an aspartic acid residue in Ad9 therefore accounts, at least in part, for the reduced CAR binding affinity of the Ad9 fiber knob. Site directed mutagenesis of CAR revealed that CAR residues Leu73 and Lys121 and/or Lys123 are critical contact residues, with Tyr80 and Tyr83 being peripherally involved in the binding interaction with the Ad5, Ad9, Ad12, and Ad41L fiber knobs. The overall affinities and the association and dissociation rate constants for wild-type CAR as well as Tyr80 and Tyr83 CAR mutants differed between the serotypes, indicating that their binding modes, although similar, are not identical. PMID- 11435606 TI - Deletion mapping of the encephalomyocarditis virus primary cleavage site. AB - The cotranslational, primary self-cleavage reaction of cardiovirus polyprotein relies on a highly conserved, short segment of amino acids at the 2A-2B protein boundary. The amino terminus of the required element for encephalomyocarditis virus has now been mapped to include Tyr(126) of the 2A protein, the 18th amino acid before the cleavage site. PMID- 11435607 TI - The specificity of interactions between nuclear hormone receptors and corepressors is mediated by distinct amino acid sequences within the interacting domains. AB - The thyroid hormone receptor (TR) and retinoic acid receptor (RAR) isoforms interact with the nuclear corepressors [NCoR (nuclear corepressor protein) and SMRT (silencing mediator for retinoid and thyroid hormone receptors)] in the absence of ligand to silence transcription. NCoR and SMRT contain C-terminal nuclear hormone receptor (NHR) interacting domains that each contain variations of the consensus sequence I/L-x-x-I/V-I (CoRNR box). We have previously demonstrated that TRbeta1 preferentially interacts with NCoR, whereas RARalpha prefers SMRT. Here, we demonstrate that this is due, in part, to the presence of a novel NCoR interacting domain, termed N3, upstream of the previously described domains. An analogous domain is not present in SMRT. This domain is specific for TR and interacts poorly with RAR. Our data suggest that the presence of two corepressor interacting domains are necessary for full interactions with nuclear receptors in cells. Interestingly, mutation of N3 alone specifically decreases binding of NCoR to TR in cells but does not decrease NCoR-RAR interactions. In addition, while the exact CoRNR box sequence of a SMRT interacting domain is critical for recruitment of SMRT by RAR, the CoRNR box sequences themselves do not explain the strong interaction of the N2 domain with TRbeta1. Additional regions distal to the CoRNR box sequence are needed for optimal binding. Thus, through sequence differences in known interacting domains and the presence of a newly identified interacting domain, NCoR is able to preferentially bind TRbeta1. These preferences are likely to be important in corepressor action in vivo. PMID- 11435608 TI - Interleukin-2 inhibits glucocorticoid receptor transcriptional activity through a mechanism involving STAT5 (signal transducer and activator of transcription 5) but not AP-1. AB - Cytokines and glucocorticoids (GCs) signaling pathways interfere with each other in the regulation of apoptosis and gene expression in the immune system. Interleukin-2 (IL-2), through the Janus kinase/signal transducers and activators of transcription (Jak/STAT) and mitogen-activated protein kinase (MAPK) pathways, activates STAT5 and activated protein-1 (AP-1) transcription factors, respectively, which are known to repress glucocorticoid receptor (GR) activity, at least in part, through protein-protein interactions. In this work, we have analyzed the mechanisms whereby IL-2 down-regulates the GC-induced transactivation of the mouse mammary tumor virus long terminal repeat (MMTV-LTR) in murine CTLL-2 T lymphocytes. Mutagenesis studies revealed that the MMTV-LTR STAT5 binding site (-923/-914) was not required for IL-2-mediated inhibition but identified both glucocorticoid response elements (GREs) and the -104/+1 region as critical elements for this negative response. The DNA binding activities of transcription factors required for GC-mediated activation of the MMTV-LTR promoter and that bind to the -104/+1 region (nuclear factor-1, Oct-1) were not affected by IL-2 treatment. Overexpression of wild-type STAT5B enhanced the effect of IL-2 on MMTV-LTR activity, and a dominant negative form of STAT5B (Y699F) abolished the IL-2-mediated MMTV-LTR inhibition, whereas AP-1 activation had no effect in this system. Direct interaction between liganded GR and STAT5 was observed in CTLL-2 cells in a STAT5 phosphorylation-independent manner. Overexpression of nuclear coactivators CBP (CREB-binding protein) or SRC-1a (steroid receptor coactivator 1a) did not blunt IL-2 inhibitory effects. We suggest that the STAT5-repressive activity on the GC-dependent transcription may involve direct interaction of STAT5 with GR, is dependent on the promoter context and STAT5 activation level, and occurs independently of coactivators levels in T cells. PMID- 11435609 TI - Dynamic changes in subcellular localization of mineralocorticoid receptor in living cells: in comparison with glucocorticoid receptor using dual-color labeling with green fluorescent protein spectral variants. AB - Mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) are ligand dependent transcription factors. Although it is generally accepted that GR is translocated into the nucleus from the cytoplasm only after ligand binding, the subcellular localization of MR is still quite controversial. We examined the intracellular trafficking of MR in living neurons and nonneural cells using a fusion protein of green fluorescent protein (GFP) and rat MR (GFP-MR). Corticosterone (CORT) induced a rapid nuclear accumulation of GFP-MR, whereas in the absence of ligand, GFP-MR was distributed in both cytoplasm and nucleus in the majority of transfected cells. Given the differential action of MR and GR in the central nervous system, it is important to elucidate how the trafficking of these receptors between cytoplasm and nucleus is regulated by ligand. To examine the simultaneous trafficking of MR and GR within single living cells, we use different spectral variants of GFP, yellow fluorescent protein (YFP) and cyan fluorescent protein (CFP), linked to MR and GR, respectively. In COS-1 cells, expressing no endogenous corticosteroid receptors, the YFP-MR chimera was accumulated in the nucleus faster than the CFP-GR chimera in the presence of 10( 9) M CORT, while there was no significant difference in the nuclear accumulation rates in the presence of 10(-6) M CORT. On the other hand, in primary cultured hippocampal neurons expressing endogenous receptors, the nuclear accumulation rates of the YFP-MR chimera and CFP-GR chimera were nearly the same in the presence of both concentrations of CORT. These results suggest that CORT-induced nuclear translocation of MR and GR exhibits differential patterns depending on ligand concentrations or cell types. PMID- 11435610 TI - Molecular identification and characterization of a and b forms of the glucocorticoid receptor. AB - The human glucocorticoid receptor (hGRalpha) is a ligand-activated transcription factor that mediates the physiological effects of corticosteroid hormones and is essential for life. Originally cloned in 1986, the transcriptionally active hGRalpha was reported to be a single protein species of 777 amino acids (molecular mass = 94 kDa). Biochemical data, obtained using various mammalian tissues and cell lines, however, have consistently revealed an additional, slightly smaller, second hGR protein (molecular mass = 91 kDa) that is not recognized by antibodies specific for the transcriptionally inactive and dominant negative, non-hormone-binding hGRbeta isoform. We report here that when a single GR cDNA is transfected in COS-1 cells, or transcribed and translated in vitro, two forms of the receptor are observed, similar to those seen in cells that contain endogenous GR. These data suggest that two forms of the hGRalpha are produced by alternative translation of the same gene and are henceforth termed GR A and GR-B. To test this hypothesis, we have investigated the role of an internal ATG codon corresponding to methionine 27 (M27) as a potential alternative translation initiation site for the GR. Mutagenesis of this ATG codon to ACG in human, rat, and mouse GR cDNA results in generation of a single 94-kDa protein species, GR-A. Moreover, mutagenesis of the initial ATG codon to ACG (Met 1 to Thr) also resulted in production of single, shorter protein species (91 kDa), GR B. Mutagenesis of the Kozak translation initiation sequence strongly indicates that a leaky ribosomal scanning mechanism is responsible for generating the GR-A and -B isoforms. Western blot analysis using peptide-specific antibodies show both the A and B receptor forms are present in human cell lines. Both receptors exhibit similar subcellular localization and nuclear translocation after ligand activation. Functional analyses of hGR-A and hGR-B under various glucocorticoid responsive promoters reveal the shorter hGR-B to be nearly twice as effective as the longer hGR-A species in gene transactivation, but not in transrepression. PMID- 11435611 TI - Engineering of a mouse for the in vivo profiling of estrogen receptor activity. AB - In addition to their well known control of reproductive functions, estrogens modulate important physiological processes. The identification of compounds with tissue-selective activity will lead to new drugs mimicking the beneficial effects of estrogen on the prevention of osteoporosis and cardiovascular or neurodegenerative diseases, while avoiding its detrimental proliferative effects. As an innovative model for the in vivo identification of new selective estrogen receptor modulators (SERMs), we engineered a mouse genome to express a luciferase reporter gene ubiquitously. The constructs for transgenesis consist of the reporter gene driven by a dimerized estrogen-responsive element (ERE) and a minimal promoter. Insulator sequences, either matrix attachment region (MAR) or beta-globin hypersensitive site 4 (HS4), flank the construct to achieve a generalized, hormoneresponsive luciferase expression. In the mouse we generated, the reporter expression is detectable in all 26 tissues examined, but is induced by 17beta-estradiol (E2) only in 15 of them, all expressing estrogen receptors (ERs). Immunohistochemical studies show that in the mouse uterus, luciferase and ERs colocalize. In primary cultures of bone marrow cells explanted from the transgenic mice and in vivo, luciferase activity accumulates with increasing E(2) concentration. E2 activity is blocked by the ER full antagonist ICI 182,780. Tamoxifen shows partial agonist activity in liver and bone when administered to the animals. In the mouse system here illustrated, by biochemical, immunohistochemical, and pharmacological criteria, luciferase content reflects ER transcriptional activity and thus represents a novel system for the study of ER dynamics during physiological fluctuations of estrogen and for the identification of SERMs or endocrine disruptors. PMID- 11435612 TI - Allosteric modulation of estrogen receptor conformation by different estrogen response elements. AB - Estrogen-regulated gene expression is dependent on interaction of the estrogen receptor (ER) with the estrogen response element (ERE). We assessed the ability of the ER to activate transcription of reporter plasmids containing either the consensus vitellogenin A2 ERE or the imperfect pS2, vitellogenin B1, or oxytocin (OT) ERE. The A2 ERE was the most potent activator of transcription. The OT ERE was significantly more effective in activating transcription than either the pS2 or B1 ERE. In deoxyribonuclease I (DNase I) footprinting experiments, MCF-7 proteins protected A2 and OT EREs more effectively than the pS2 and B1 EREs. Limited protease digestion of the A2, pS2, B1, or OT ERE-bound receptor with V8 protease or proteinase K produced distinct cleavage products demonstrating that individual ERE sequences induce specific changes in ER conformation. Receptor interaction domains of glucocorticoid receptor interacting protein 1 and steroid receptor coactivator 1 bound effectively to the A2, pS2, B1, and OT ERE-bound receptor and significantly stabilized the receptor-DNA interaction. Similar levels of the full-length p160 protein amplified in breast cancer 1 were recruited from HeLa nuclear extracts by the A2, pS2, B1, and OT ERE-bound receptors. In contrast, significantly less transcriptional intermediary factor 2 was recruited by the B1 ERE-bound receptor than by the A2 ERE-bound receptor. These studies suggest that allosteric modulation of ER conformation by individual ERE sequences influences the recruitment of specific coactivator proteins and leads to differential expression of genes containing divergent ERE sequences. PMID- 11435613 TI - Regulation of gene Expression by 1alpha,25-dihydroxyvitamin D3 and Its analog EB1089 under growth-inhibitory conditions in squamous carcinoma Cells. AB - Analogs of 1alpha,25-dihydroxyvitamin D(3) (1alpha, 25(OH)2D3) inhibit growth in vitro and in vivo of cells derived from a variety of tumors. Here, we examined the effects of 1alpha,25(OH)2D3 and its analog EB1089 on proliferation and target gene regulation of human head and neck squamous cell carcinoma (SCC) lines SCC4, SCC9, SCC15, and SCC25. A range of sensitivities to 1alpha,25(OH)2D3 and EB1089 was observed, from complete G0/G1 arrest of SCC25 cells to only 50% inhibition of SCC9 cell growth. All lines expressed similar levels of vitamin D3 receptor (VDR) mRNA and protein, and no significant variation was observed in 1alpha,25(OH)2D3 dependent induction of the endogenous 24-hydroxylase gene, or of a transiently transfected 1alpha,25(OH)2D3-sensitive reporter gene. The antiproliferative effects of 1alpha,25(OH)2D3 and EB1089 in SCC25 cells were analyzed by screening more than 4,500 genes on two cDNA microarrays, yielding 38 up-regulated targets, including adhesion molecules, growth factors, kinases, and transcription factors. Genes encoding factors implicated in cell cycle regulation were induced, including the growth arrest and DNA damage gene, gadd45alpha, and the serum- and glucocorticoid-inducible kinase gene, sgk. Induction of GADD45alpha protein in EB1089-treated cells was confirmed by Western blotting. Moreover, while expression of proliferating cell nuclear antigen (PCNA) was reduced in EB1089 treated cells, coimmunoprecipitation studies revealed increased association between GADD45alpha and PCNA in treated cells, consistent with the capacity of GADD45alpha to stimulate DNA repair. While 1alpha,25(OH)2D3 and EB1089 modestly induced transcripts encoding the cyclin-dependent kinase inhibitor p21(waf1/cip1), no changes in protein levels were observed, indicating that p21(waf1/cip1) induction does not contribute to the antiproliferative effects of 1alpha,25(OH)2D3 and EB1089 in SCC cells. Finally, in partially resistant SCC9 cells, there was extensive loss of target gene regulation (10 of 10 genes tested), indicating that resistance arises from widespread loss of 1alpha,25(OH)2D3-dependent gene regulation in the presence of normal levels of functional VDRs. PMID- 11435614 TI - Reconstruction of ligand-dependent transactivation of Choristoneura fumiferana ecdysone receptor in yeast. AB - Ecdysteroids play an important role in regulating development and reproduction in insects. Interaction of 20-hydroxyecdysone (20E) with ecdysone receptor (EcR) as a heterodimer with ultraspiracle (USP) protein triggers the activation of 20E responsive genes. In this paper we describe a ligand-mediated transactivation system in yeast using the spruce budworm Choristoneura fumiferana ecdysone receptor (CfEcR). Coexpression of C. fumiferana USP (CfUSP) with CfEcR in yeast led to constitutive transcription of the reporter gene. However, deletion of the A/B domain of CfUSP abolished constitutive activity observed for the CfUSP:CfEcR complex. Replacement of USP with its mammalian homolog retinoid X receptors (RXRs) abolished the constitutive activity of the heterodimer but it did not restore EcR ligand-mediated transactivation. These data suggest that USP and its A/B domain play a role in the constitutive function of CfEcR:USP in yeast. A ligand-mediated transactivation was observed when GRIP1, a mouse coactivator gene, was added to EcR:RXR or EcR:DeltaA/BUSP complexes. Deletion of the A/B domain of EcR in the context of DeltaA/BEcR:RXR:GRIP1 or DeltaA/BEcR:DeltaA/BUSP:GRIP1 dramatically improved the ligand-dependent transactivation. This is the first example of highly efficient ligand-dependent transactivation of insect EcR in yeast. Analysis of transactivation activity of different ecdysteroidal compounds showed that the yeast system remarkably mimics the response observed in insect tissue culture cells and whole insect systems. The results open the way to develop assays that can be used to screen novel species-specific ecdysone agonist/antagonist insecticides. PMID- 11435615 TI - Autonomous rexinoid death signaling is suppressed by converging signaling pathways in immature leukemia cells. AB - On their own, retinoid X receptor (RXR)-selective ligands (rexinoids) are silent in retinoic acid receptor (RAR)-RXR heterodimers, and no selective rexinoid program has been described as yet in cellular systems. We report here on the rexinoid signaling capacity that triggers apoptosis of immature promyelocytic NB4 cells as a default pathway in the absence of survival factors. Rexinoid-induced apoptosis displays all features of bona fide programmed cell death and is inhibited by RXR, but not RAR antagonists. Several types of survival signals block rexinoid-induced apoptosis. RARalpha agonists switch the cellular response toward differentiation and induce the expression of antiapoptosis factors. Activation of the protein kinase A pathway in the presence of rexinoid agonists induces maturation and blocks immature cell apoptosis. Addition of nonretinoid serum factors also blocks cell death but does not induce cell differentiation. Rexinoid-induced apoptosis is linked to neither the presence nor stability of the promyelocytic leukemia-RARalpha fusion protein and operates also in non-acute promyelocytic leukemia cells. Together our results support a model according to which rexinoids activate in certain leukemia cells a default death pathway onto which several other signaling paradigms converge. This pathway is entirely distinct from that triggered by RAR agonists, which control cell maturation and postmaturation apoptosis. PMID- 11435616 TI - Isoform-specific transcriptional regulation by thyroid hormone receptors: hormone independent activation operates through a steroid receptor mode of co-activator interaction. AB - Thyroid hormone receptors (T3Rs) are hormone-regulated transcription factors that play important roles in vertebrate homeostasis, differentiation, and development. T3Rs are synthesized as multiple isoforms that display tissue-specific expression patterns and distinct transcriptional properties. Most T3R isoforms associate with co-activator proteins and mediate transcriptional activation only in the presence of thyroid hormone. The pituitary-specific T3Rbeta-2 isoform departs from this general rule and is able to interact with p160 co-activators, and to mediate transcriptional activation in both the absence and presence of hormone. We report here that this hormone-independent activation is mediated by contacts between the unique N terminus of T3Rbeta-2 and an internal interaction domain in the SRC-1 (steroid receptor co-activator-1) and GRIP-1 (glucocorticoid receptor interacting protein 1) co-activators. These hormone-independent contacts between T3Rbeta-2 and the p160 co-activators are distinct in sequence and function from the LXXLL motifs that mediate hormone-dependent transcriptional activation and resemble instead a mode of co-activator recruitment previously observed only for the steroid hormone receptors and only in the presence of steroid hormone. Our results suggest that the transcriptional properties of the different T3R isoforms represent a combinatorial mixture of repression, antirepression, and hormone independent and hormone-dependent activation functions that operate in conjunction to determine the ultimate transcriptional outcome. PMID- 11435617 TI - Analysis of parathyroid hormone (PTH)/secretin receptor chimeras differentiates the role of functional domains in the pth/ pth-related peptide (PTHrP) receptor on hormone binding and receptor activation. AB - The type 1 parathyroid hormore receptor (PTH1r) belongs to the class II family of G protein-coupled receptors. To delineate the sites in the PTH1r's N-terminal region, and the carboxy-core domain (transmembrane segments + extracellular loops) involved in PTH binding, we have evaluated the functional properties of 27 PTH1-secretin chimeras receptors stably expressed in HEK-293 cells. The wild type and chimeric receptors were analyzed for cell surface expression, binding for PTH and secretin, and functional responsiveness (cAMP induction) toward secretin and PTH. The expression levels of the chimeric receptors were comparable to that of the PTH1r (60-100%). The N-terminal region of PTH1r was divided into three segments that were replaced either singly or in various combinations with the homologous region of the secretin receptor (SECr). Substitution of the carboxy terminal half (residues 105-186) of the N-terminal region of PTH1r for a SECr homologous segment did not reduced affinity for PTH but abolished signaling in response to PTH. This data indicate that receptor activation is dissociable from high affinity hormone binding in the PTH1r, and that the N-terminal region might play a critical role in the activation process. Further segment replacements in the N-termini focus on residues 105-186 and particularly residues 146-186 of PTH1r as providing critical segments for receptor activation. The data obtained suggest the existence of two distinct PTH binding sites in the PTH1r's N-terminal region: one site in the amino-terminal half (residues 1-62) (site 1) that participates in high-affinity PTH binding; and a second site of lower affinity constituted by amino acid residues scattered throughout the carboxy-terminal half (residues 105-186) (site 2). In the absence of PTH binding to site 1, higher concentrations of hormone are required to promote receptor activation. In addition, elimination of the interaction of PTH with site 2 results in a loss of signal transduction without loss of high-affinity PTH binding. Divers substitutions of the extracellular loops of the PTH1r highlight the differential role of the first- and third extracellular loop in the process of PTH1r activation after hormone binding. A chimera containing the entire extracellular domains of the PTH1r and the transmembrane + cytoplasmic domains of SECr had very low PTH binding affinity and did not signal in response to PTH. Further substitution of helix 5 of PTH1r in this chimera increased affinity for PTH that is close to the PTH affinity for the wild-type PTH1r but surprisingly, did not mediate signaling response. Additional substitutions of PTH1r's helices in various combinations emphasize the fundamental role of helix 3 and helix 6 on the activation process of the PTH1r. Overall, our studies demonstrated that several PTH1r domains contribute differentially to PTH binding affinity and signal transduction mechanism and highlight the role of the N-terminal domain and helix 3 and helix 6 on receptor activation. PMID- 11435618 TI - Maturity-onset diabetes of the young Type 1 (MODY1)-associated mutations R154X and E276Q in hepatocyte nuclear factor 4alpha (HNF4alpha) gene impair recruitment of p300, a key transcriptional co-activator. AB - Hepatocyte nuclear factor 4alpha (HNF4alpha) is a nuclear receptor involved in glucose homeostasis and is required for normal beta-cell function. Mutations in the HNF4alpha gene are associated with maturity-onset diabetes of the young type 1. E276Q and R154X mutations were previously shown to impair intrinsic transcriptional activity (without exogenously supplied co-activators) of HNF4alpha. Given that transcriptional partners of HNF4alpha modulate its intrinsic transcriptional activity and play crucial roles in HNF4alpha function, we investigated the effects of these mutations on potentiation of HNF4alpha activity by p300, a key co-activator for HNF4alpha. We show here that loss of HNF4alpha function by both mutations is increased through impaired physical interaction and functional cooperation between HNF4alpha and p300. Impairment of p300-mediated potentiation of HNF4alpha transcriptional activity is of particular importance for the E276Q mutant since its intrinsic transcriptional activity is moderately affected. Together with previous results obtained with chicken ovalbumin upstream promoter-transcription factor II, our results highlight that impairment of recruitment of transcriptional partners represents an important mechanism leading to abnormal HNF4alpha function resulting from the MODY1 E276Q mutation. The impaired potentiations of HNF4alpha activity were observed on the promoter of HNF1alpha, a transcription factor involved in a transcriptional network and required for beta-cell function. Given its involvement in a regulatory signaling cascade, loss of HNF4alpha function may cause reduced beta cell function secondary to defective HNF1alpha expression. Our results also shed light on a better structure-function relationship of HNF4alpha and on p300 sequences involved in the interaction with HNF4alpha. PMID- 11435619 TI - Functional expression and characterization of a voltage-gated CaV1.3 (alpha1D) calcium channel subunit from an insulin-secreting cell line. AB - L-type calcium channels mediate depolarization-induced calcium influx in insulin secreting cells and are thought to be modulated by G protein-coupled receptors (GPCRs). The major fraction of L-type alpha1-subunits in pancreatic beta-cells is of the neuroendocrine subtype (CaV1.3 or alpha1D). Here we studied the biophysical properties and receptor regulation of a CaV1.3 subunit previously cloned from HIT-T15 cells. In doing so, we compared this neuroendocrine CaV1.3 channel with the cardiac L-type channel CaV1.2a (or alpha1C-a) after expression together with alpha2delta- and beta3-subunits in Xenopus oocytes. Both the current voltage relation and voltage dependence of inactivation for the neuroendocrine CaV1.3 channel were shifted to more negative potentials compared with the cardiac CaV1.2 channel. In addition, the CaV1.3 channel activated and inactivated more rapidly than the CaV1.2a channel. Both subtypes showed a similar sensitivity to the dihydropyridine (+)isradipine. More interestingly, the CaV1.3 channels were found to be stimulated by ligand-bound G(i)/G(o)-coupled GPCRs whereas a neuronal CaV2.2 (or alpha1B) channel was inhibited. The observed receptor-induced stimulation of CaV1.3 channels could be mimicked by phorbol-12 myristate-13-acetate and was sensitive to inhibitors of protein kinases, but not to the phosphoinositol-3-kinase-inhibitor wortmannin, pointing to serine/threonine kinase-dependent regulation. Taken together, we describe a neuroendocrine L-type CaV1.3 calcium channel that is stimulated by G(i)/G(o) coupled GPCRs and differs significantly in distinct biophysical characteristics from the cardiac subtype (CaV1.2a), suggesting that the channels have different roles in native cells. PMID- 11435620 TI - Differential expression of ionic channels in rat anterior pituitary cells. AB - Secretory anterior pituitary cells are of the same origin, but exhibit cell type specific patterns of spontaneous intracellular Ca2+ signaling and basal hormone secretion. To understand the underlying ionic mechanisms mediating these differences, we compared the ionic channels expressed in somatotrophs, lactotrophs, and gonadotrophs from randomly cycling female rats under identical cell culture and recording conditions. Our results indicate that a similar group of ionic channels are expressed in each cell type, including transient and sustained voltage-gated Ca2+ channels, tetrodotoxin-sensitive Na+ channels, transient and delayed rectifying K+ channels, and multiple Ca2+ -sensitive K+ channel subtypes. However, there were marked differences in the expression levels of some of the ionic channels. Specifically, lactotrophs and somatotrophs exhibited low expression levels of tetrodotoxin-sensitive Na+ channels and high expression levels of the large-conductance, Ca2+ -activated K+ channel compared with those observed in gonadotrophs. In addition, functional expression of the transient K+ channel was much higher in lactotrophs and gonadotrophs than in somatotrophs. Finally, the expression of the transient voltage-gated Ca2+ channels was higher in somatotrophs than in lactotrophs and gonadotrophs. These results indicate that there are cell type-specific patterns of ionic channel expression, which may be of physiological significance for the control of Ca2+ homeostasis and secretion in unstimulated and receptor-stimulated anterior pituitary cells. PMID- 11435621 TI - Endocrine-related resources from the National Institutes of Health. PMID- 11435623 TI - Jaundice in older children and adolescents. PMID- 11435624 TI - Preparticipation examination of the adolescent athlete: part 2. PMID- 11435625 TI - Visual diagnosis: an adolescent female who has increasing hair growth. PMID- 11435626 TI - Index of suspicion. Case 1. Diagnosis: TB meningitis. PMID- 11435627 TI - Obesity. PMID- 11435628 TI - Human gene therapy: are we still expecting too much, too soon? PMID- 11435629 TI - The use of capture-recapture techniques in determining the prevalence of type 2 diabetes. PMID- 11435630 TI - Maternal and neonatal outcomes in pregestational and gestational diabetes mellitus, and the influence of maternal obesity and weight gain: the DEPOSIT study. Diabetes Endocrine Pregnancy Outcome Study in Toronto. AB - We prospectively studied pregnancy outcome in 428 women with gestational diabetes mellitus (DM) and 196 women with pregestational DM, with particular reference to the influence of maternal obesity and excessive weight gain. These were consecutive singleton pregnancies delivered in our institution over 5 years. After controlling for multiple risk factors, including maternal BMI and pregnancy weight gain, women with pregestational DM were at increased risk (compared to those with gestational DM) for Caesarean delivery (OR 3.6, 95%CI 2.3-5.6), shoulder dystocia or cephalopelvic disproportion (OR 2.2, 95%CI 1.3-3.6), and gestational hypertension or toxaemia (OR 3.0, 95%CI 1.7-5.4). The offspring of these women were also at increased risk for admission to the neonatal intensive care unit (OR 4.0, 95%CI 2.3-6.8), large-for-gestational-age birthweight (OR 3.5, 95%CI 2.2-5.6), and preterm birth before 37 weeks (OR 3.8, 95%CI 2.5-5.9). Maternal obesity, and, to a lesser degree, excessive weight gain, were also independent risk factors for all these adverse maternal and neonatal outcomes, regardless of the type of DM, except for shoulder dystocia/cephalopelvic disproportion. PMID- 11435631 TI - Endemic strongyloidiasis on the Spanish Mediterranean coast. AB - Diagnosis and treatment of Strongyloides stercoralis infection can be difficult, and a high degree of clinical suspicion in patients who have visited an endemic area is required. We describe the epidemiology and clinical features of 152 prospectively identified cases of strongyloidiasis in an European region, and identify risk factors for the development of severe forms of the disease. This was a prospective study of all patients admitted to a single institution over an 8-year period. Patients (n=152) were mainly elderly male farmers (79%) who had acquired the disease by working barefoot in contact with soil and ingesting non drinking water. Eosinophilia was a sensitive marker for the infection (82%). Twenty patients (13%) developed severe forms of the illness and six patients (4%) died. A significant association was found between severe forms of strongyloidiasis and steroid usage (OR 9.0, 95%CI 2.1-37.6), immunodebilitating illness (OR 10.1, 95%CI 3.2-32.3) and other immunosuppressive therapy (OR 13.7, 95%CI 2.9-58.7), but by logistic regression analysis, only immunodebilitating disease was as a risk factor (OR 2.1, 95%CI 1.78-2.43). S. stercolaris infection is endemic in the Spanish Mediterranean coast. The frequent development of severe forms of the disease, with a high mortality, makes early recognition and treatment essential. PMID- 11435632 TI - Cost-effectiveness of screening and extended anticoagulation for carriers of both factor V Leiden and prothrombin G20210A. AB - Carriers of a double thrombophilic mutation (factor V Leiden and prothrombin G20210A) are at high risk of a recurrent venous thromboembolism (VTE), and may benefit from a longer course of secondary prophylaxis. We examined the costs and health benefits of screening for both the mutations, provided that double heterozygotes undergo 2 years of anticoagulation as compared to the standard 6 months. We thus pooled the available evidence and calculated that the OR for recurrence in double heterozygotes was 5.9 (95% CI 2.65-13.20). A Markov model tracked patients' health lifelong, and calculated that prolonged prophylaxis saved 26 quality-adjusted days of life and $410 per double heterozygote treated. Screening all the patients with venous thromboembolism thus provided one additional day of life at the cost of 13624 $/QALY (95% CI 12 965-22 889). Screening was not cost-effective in those cohorts with a low prevalence of the mutations, a high bleeding risk or in those where prophylaxis prevented <65% of recurrences. Screening for factor V Leiden and prothrombin G20210A, with prolonged prophylaxis of double carriers, is cost-effective in most patients with VTE. PMID- 11435633 TI - Antimicrobial management of acute exacerbation of chronic airflow limitation. AB - Antibiotics are frequently administered for exacerbations of chronic obstructive pulmonary disease and asthma, yet their role remains unclear. We prospectively audited the antimicrobial management of 167 patients aged >50 years hospitalized for exacerbations of chronic airflow limitation. Antibiotics were commenced on admission for 151 (90%) patients (oral 52%, intravenous 38%), including 17/23 (74%) with no evidence of fever, purulent sputum, leucocytosis or inflammatory chest X-ray changes. The mean number of different antibiotics prescribed was 1.8 (range 0-6); a wide range of antibiotics and antibiotic combinations were used. Sputum samples were sent for microbiological examination in 101 (61%) patients. Sputum culture was positive in 34, but only 11 (7% of the total) had amoxycillin resistant organisms in their sputum. Seventeen patients (10%) developed diarrhoea while in hospital. Under logistic regression analysis, total number of antibiotics prescribed (p<0.0001) and age (p=0.0062) were the two factors associated with hospital-acquired diarrhoea. Only 34% of patients had received an influenza vaccination in the winter of the study, and 10% a pneumococcal vaccination within the last 5 years. In routine clinical practice, aggressive antibiotic therapy was frequently administered to patients admitted with chronic airflow limitation, despite limited clinical, radiological and microbial indications. Excessive use of antibiotics has important implications, including morbidity (antibiotic-associated diarrhoea), cost and the potential for increased microbial antibiotic resistance. A minority of patients with chronic airflow limitation are being vaccinated against influenza and Pneumococcus. PMID- 11435634 TI - Recently diagnosed sexually HIV-infected patients: seroconversion interval, partner notification period and a high yield of HIV diagnoses among partners. AB - We collected data on the outcome of partner notification (PN) for 356 index patients (iPs) newly diagnosed with HIV, all sexually infected, in six European countries, 1995-96. The PN period (the period during which exposed partners should be notified about their risk of being HIV infected) could be defined on the basis of last negative HIV test in 170/356 (48%), from other key dates for 12, and for 14 more on the sole basis of known HIV exposure (total 196/356, 55%). However, of the 170 iPs with a last negative HIV test date, PN period was not defined in 52 (31%), even though 28/52 were recent seroconverters (negative HIV test within 24 months of HIV diagnosis). Discrepancies in PN period evaluation were common: of the 85 iPs for whom PN period was reportedly defined as '3 months before last HIV-negative date', the period actually used was equal to this in only 12. A user-friendly worksheet would simplify the task of eliciting and using key dates to define the PN period. Forms on PN outcome were received for 166/200 (83%) reported current partners, but only 124/508 (24%) ex-partners. Fifty-two of 290 (18%) notified partners already knew they were HIV-positive, and 38/130 (29%) notified partners for whom pre-PN status was negative or unknown were HIV positive on post-PN testing. Results of HIV tests (if any) after PN were unknown for 37/133 (28%) current partners not already known to be HIV-positive, and for 71/105 (68%) ex-partners-a doubling of the information loss rate. Notification of 133 current and 105 ex-partners not previously identified as HIV-positive thus produced 28 (21%) and 10 (10%) new HIV diagnoses, respectively. The total yield of HIV diagnoses was 38/100 (38%) partners of recent seroconverters (22 already known plus 16 PN-diagnosed), compared with 52/190 (27%) partners of other iPs (30 already known plus 22 PN-diagnosed). We propose prioritization for PN of partners of recent seroconverters, augmented surveillance for HIV diagnoses and recurrence in PN networks of at-risk, non-notified partners, and new measures that preserve anonymity to alert potential and past sexual contacts of HIV-diagnosed individuals who are unable or unwilling to assist in PN. PMID- 11435635 TI - Ethnic differences in hypertension and blood pressure control in the UK. PMID- 11435636 TI - Evidence-based diagnosis of severe aortic stenosis. PMID- 11435637 TI - Predictors of re-occlusion after successful recanalization of chronic total occlusion. AB - The objective of this study was to highlight the incidence and predictors of re occlusion after successful recanalization of chronic total coronary occlusions. METHODS AND RESULTS: Following successful recanalization and stent implantation in 716 coronary lesions (665 patients) with chronic total occlusion, four hundred and five (56.6%) lesions (375 patients) underwent repeat angiography within 6 months. Restenosis (> or = 50% lumen narrowing) was observed in 151 (37.3%) lesions; forty-three (10.6%) of these lesions had complete re-occlusion and constituted the study population. In this group, final angiographic minimal lumen diameter (MLD) was 2.6 +/- 0.51 mm and final percent diameter stenosis was 18 +/- 11. Univariate analysis revealed significant correlation between re-occlusion and restenotic lesions, final balloon diameter, final percent diameter stenosis, final angiographic MLD, number of stents per lesion and total stent length. By multivariate analysis, the only independent predictor of re-occlusion was total stent length (OR = 1.46, 95% CI = 1.12-1.82; p = 0.0069). CONCLUSION: Re occlusion occurs in about 11% of cases after stenting chronic total occlusion. The most important predictor of re-occlusion seems to be stent length. PMID- 11435638 TI - No jacket required. PMID- 11435639 TI - Intravascular ultrasound analysis of arterial remodeling in the double-injury swine coronary artery model of restenosis. AB - BACKGROUND: Vascular remodeling is a major component of atherosclerotic and restenotic processes. The aim of this study was to evaluate remodeling at two different axial loci in a restenotic (double-injury) coronary artery model in the hypercholesterolemic minipig. METHODS: The cross-sectional area (CSA) of the lumen, artery and neointima following a single stenotic injury (SI) and second restenotic injury (RI) was measured by serial intravascular ultrasound (IVUS). IVUS CSAs were determined before, immediately after and 4 weeks after each injury. Analyses were performed at two selected axial loci, defined as: 1) locus of the minimal luminal area (MLA) 4 weeks after SI (measured immediately prior to the RI); and 2) locus of the MLA at 4 weeks after RI. Loci were selected to mimic diverse approaches used in studies of remodeling. RESULTS: Restenotic remodeling, defined by locus 1, was unchanged 4 weeks following the RI and neointima formation accounted for the chronic lumen loss. In contrast, constrictive remodeling was the prominent arterial response defined by locus 2. Regardless of the MLA analysis, artery wall enlargement occurred in more than half of the pigs at 4 weeks following the SI. Balloon/artery ratios were comparable during the two injuries and could not account for observed differences in remodeling. Remodeling results obtained by comparison of artery CSA to a reference CSA differed from serial IVUS analysis. CONCLUSIONS: In the presence of a pre-established stenosis, vascular remodeling depends on the axial locus of interest. In contrast, remodeling following injury of a naive artery is independent of defined axial locations used here and is predominantly one of outward remodeling. PMID- 11435640 TI - Percutaneous transluminal septal myocardial ablation in drug-resistant hypertrophic obstructive cardiomyopathy: 18-month follow-up results. AB - OBJECTIVE: Surgery has been the only therapeutic option in patients with hypertrophic obstructive cardiomyopathy (HOCM) who are resistant to standard treatment. Percutaneous transluminal septal myocardial ablation (PTSMA) by alcohol-induced occlusion of the septal artery for the reduction of left ventricular outflow tract (LVOT) gradient is a novel method. However, long-term clinical follow-up is insufficient. This study reports the acute and mid-term results after PTSMA in symptomatic patients with HOCM. METHODS: Six patients (4 men and 2 women) with symptomatic HOCM and inadequate response to pharmacologic therapy had their major septal branches ablated with alcohol. Mean duration of follow-up was 1.5 years. RESULTS: There was reduction in LVOT gradient in all patients (100%) with a mean reduction of 50 mmHg. Two patients developed complete heart block (CHB) and 3 patients developed bifascicular block. CHB persisted in 1 patient. Two patients died within 8 days of the procedure; one due to cerebrovascular accident and 1 due to asystole. The patient who died of asystole had CHB but refused permanent pacemaker implantation. CHB reverted in 1 patient and bifascicular blocks persisted in 3 patients. Clinical and echocardiographic follow-up was achieved in the 4 surviving patients after 1.5 years. All surviving patients had LVOT gradient reduction and clinical improvement. CONCLUSION: PTSMA for HOCM is a non-surgical technique for reducing LVOT gradient. Heart blocks are one potential complication, and may require pacemaker implantation. Long-term follow-up and a larger series of patients are required to determine conclusive therapeutic significance. PMID- 11435641 TI - Acute stent thrombosis visualized by intravascular ultrasound. AB - Subacute stent thrombosis is currently a rare but feared complication in patients undergoing coronary intervention. Intravascular ultrasound is a useful technique to guide stent implantation. However, its value in patients suffering from acute stent thrombosis has not been described. Herein, we present the intravascular ultrasound findings of a patient experiencing ongoing stent thrombosis and impending vessel closure. PMID- 11435642 TI - Diffuse alveolar hemorrhage after clopidogrel use. AB - Pulmonary hemorrhage following percutaneous coronary intervention is a very rare complication that has been previously reported in association with abciximab and tirofiban, but not with clopidogrel. We report the case of a 56-year-old man who developed diffuse alveolar hemorrhage after receiving clopidogrel for placement of a coronary stent. Since clopidogrel is a commonly used medication, clinicians should be aware of this potentially life-threatening complication. PMID- 11435643 TI - Extensive thrombus prior to elective percutaneous coronary intervention. PMID- 11435645 TI - Clinical utility of the cutting balloon. PMID- 11435646 TI - Help in the middle. PMID- 11435647 TI - Should ablative therapy be first-line therapy for idiopathic atrial fibrillation? PMID- 11435648 TI - Acute myocardial infarction due to a regressed giant coronary aneurysm as possible sequela of Kawasaki disease. AB - We report on a young patient admitted with an acute myocardial infarction due to a regressed coronary aneurysm who was treated with balloon angioplasty. The wall morphology of the coronary aneurysm using intravascular ultrasound imaging closely resembled that occurring after Kawasaki disease (KD), although our patient had no obvious past history of KD, suggesting so-called atypical KD. PMID- 11435649 TI - Animal experimentation and research in India. PMID- 11435650 TI - Combined use of isoflurane and sodium nitroprusside during active rewarming on cardiopulmonary bypass: a prospective, comparative study. AB - AIMS: To evaluate and compare the effect of isoflurane, sodium nitroprusside (SNP) and combined use of isoflurane and SNP on body rewarming and haemodynamic stability during active rewarming on cardiopulmonary bypass (CPB). SUBJECTS AND METHODS: In a prospective, randomised study 75 adult patients scheduled for coronary artery bypass grafting (CABG) under CPB were studied in three groups of 25 patients each. During active rewarming, patients of group I received SNP infusion in CPB, group-II received isoflurane through vaporiser in gas circuit of the CPB machine and group III received a combination of isoflurane inhalation (0.2-0.5%) + SNP in low doses (<1mg/kg/min). RESULTS: Mean requirements of SNP to achieve maximum pump flow during rewarming were 1.48 -/+ 0.65 mg/kg/min (range 0.3-3.5 mg/kg/min) in group I and 0.75 -/+ 0.25 mg/kg/min (range 0.2-0.85 mg/kg/min) in group III. Mean isoflurane concentration required to achieve maximum pump flow during rewarming was 0.95 -/+ 0.35% (range 0.2-1.5%) in group II and 0.35 -/+ 0.1 (range 0-0.4%) in group III. The requirements of SNP and isoflurane in group III were significantly less than group I and II (p<0.001). The haemodynamic stability was better in SNP + isoflurane group with significantly lesser requirement of inotropes. Four-scaled assessment for rewarming evaluation failed to show significant statistical difference amongst the groups. CONCLUSIONS: All three drug regimens were equally effective in terms of uniform rewarming of the body on CPB. However, combined use of SNP and isoflurane in low doses provides haemodynamic stability during CPB and is superior to either drug alone. PMID- 11435651 TI - Antiphospholipid antibody positive young stroke: an analysis of 12 cases. AB - AIMS AND OBJECTIVES: To study clinico-investigative profile of 12 young (<45 years) patients with stroke who tested positive for anti phospholipid antibodies (APLA). SUBJECTS AND METHODS: The diagnostic, clinical, laboratory and radiologic features in 12 APLA positive young patients who presented with stroke were studied. The APLA analysis included estimation of anticardiolipin (aCL) antibodies and lupus anticoagulant (LA). Other relevant tests included anti nuclear antibody, human immunodeficiency virus, Venereal Diseases Research Laboratory, platelet count, echocardiography and carotid Doppler. APLA positive strokes were those cases where either the immunoglobulin G (IgG) and immunoglobulin M (IgM) were raised or LA was positive, and other known causes were excluded. RESULTS: Levels of IgG (aCL) was raised in 11 cases (mild 7, moderate 1, high 3), IgM was elevated in all the 12 cases (moderate 2, high 10). Of the two LA positive cases both were IgM positive but in one IgG was negative. Five patients showed small multiple bilateral cerebral infarcts on computerised tomography (CT) scan. 5 patients had history of recurrent strokes. Hemiparesis was more frequent than hemiplegia. None presented with dense hemiplegia. All patients recovered to normal functional capacity and did not have recurrence on drugs. CONCLUSION: A preliminary study on APLA positive young strokes showed certain clinical and radiological features, mild to moderate stroke, pre treatment recurrences, multiple smaller infarcts on CT, which could be clustered in a subgroup of stroke in young. Incidentally these patients showed a good prognosis in terms of long term outcome. PMID- 11435652 TI - Reducing the number of daily measurements results in poor estimation of diurnal variability of peak expiratory flow in healthy individuals. AB - AIM: To determine the effect of reducing number of daily measurements on estimation of diurnal variability (DV) of peak expiratory flow (PEF). SUBJECTS AND METHODS: PEF was recorded five times daily for three days in 152 healthy adults. Amplitude percent mean (A%M) and standard deviation percent mean (SD%M) were calculated on third day from five, four, three and two daily readings. Proportion of variability explained by partial schedules was calculated and limits of agreement derived to assess if these methods could be used interchangeably. RESULTS: Four, three and two measurements explained 90-95%, 70 82% and 55% DV respectively using A%M. All schedules of partial measurement using SD%M explained >90% DV. Limits of agreement for A%M and SD%M widened as number of measurements were reduced. CONCLUSIONS: DV obtained by fewer daily measurements agrees poorly with results obtained from five measurements. SD%M is a better alternative if DV is assessed from fewer readings. PMID- 11435653 TI - Modified mesh rectopexy: a study. AB - AIM: To evaluate the efficacy of modified mesh rectopexy for complete rectal prolapse. SUBJECT AND METHODS: In a prospective study between 1989-1998, 47 patients (25 males and 22 females) underwent modified mesh rectopexy using a "Cross-shaped" knitted monofilament polypropylene. They were followed up for a period of four years postoperatively. RESULTS: Anatomical repair was achieved in all patients. Preoperative constipation, a complaint in 22 patients, was relieved in 13 patients and need for laxatives decreased in another four. There were no new cases of constipation. Sexual functions were not hampered irrespective of gender. The complications included prolonged ileus (4 patients), faecal impaction (1), partial mucosal prolapse (2) and post-operative obstruction (2). There was no recurrence. CONCLUSION: This technique aligns the rectum, avoids excessive mobilisation and division of lateral ligaments thus preventing constipation and preserving potency. We recommend this technique for patients with complete rectal prolapse with up to grade 1, 2 and 3 incontinence based on Browning and Parks classification. PMID- 11435654 TI - The cerebro-costo-mandibular syndrome: 9-year follow-up of a case. AB - Cerebro-costo-mandibular syndrome (CCMS) is a rare multiple congenital anomaly with a low survival rate. There are few reports of long-term survival in this condition. We describe the findings and management of a 9-year-old survivor of CCMS, outline the importance of early intervention and multidisciplinary team approach. The child presented in the neonatal period in respiratory distress with classical features of the syndrome. Aggressive initial respiratory management was later followed up with an integrated multidisciplinary team approach. He has been carefully followed up for nine years now, illustrating well, the course of the syndrome. PMID- 11435655 TI - Primary angiitis of the central nervous system: an ante-mortem diagnosis. AB - A rare case of primary angiitis of the central nervous system (PACNS) is reported with its clinical and magnetic resonance imaging (MRI) features. A 20-year-old girl presented with headache, projectile vomiting, unsteadiness of gait and urgency of micturition. She had left seventh nerve upper motor neuron type paresis, increased tone in all four limbs, exaggerated deep tendon reflexes, cerebellar signs, and papilloedema. Cerebrospinal fluid showed lymphocytosis with elevated protein and normal glucose level. Cerebral computerised tomographic scan and MRI showed bilateral diffuse asymmetric supra- and infra-tentorial lesions (predominantly in the supratentorial and left cerebrum). On MRI, the lesions were hyperintense on T2, and proton density-weighted images and hypointense on T1 weighted images. Based on the clinical findings of raised intracranial tension and MRI features, initial diagnoses of gliomatosis cerebrii, tuberculous meningitis, primary central nervous system lymphoma and chronic viral encephalitis were considered. PACNS was not included in the initial differentials and, an open brain biopsy was advised which established the definitive diagnosis. PMID- 11435656 TI - Cutaneous histiocytic lesions: a clinical dilemma. AB - The diagnosis of malignant histiocytosis requires a high index of clinical suspicion, awareness of its atypical features and availability of various tissue samples for morphological and special studies. The case reported here highlights the diagnostic difficulties encountered in a patient diagnosed as malignant histiocytosis who presented with cutaneous lesions in multiple foci, which included the face, groin and forearm. Only after repeated biopsies and special stains, a diagnosis of malignant histiocytosis was arrived at. Chemotherapy with CHOP regimen (cyclophosphamide, doxorubicin, vincristine and prednisone) was initiated. The response to chemotherapy was good and the patient is doing well eleven months after initial diagnosis. PMID- 11435657 TI - Cerebellar atrophy in an epileptic child: is it due to phenytoin? AB - A four and half year old epileptic child on phenytoin therapy since one year presented with signs of cerebellar dysfunction. Serum phenytoin level was high (33 mcg/ml) and computerised tomographic scan of the brain showed severe generalised cerebellar atrophy. The cerebellar signs represented drug over dosage and toxicity and persisted long after omission of phenytoin. PMID- 11435658 TI - The art and science of presentation: 35-mm slides. PMID- 11435659 TI - Images in medicine: Hashimoto's thyroiditis with orbitopathy and dermopathy. PMID- 11435660 TI - Images in pathology: pseudomyxoma peritonei masquerading as abdominal hydatidosis. PMID- 11435661 TI - Images in radiology: complete hydatidiform mole with live pregnancy in a twin gestation. PMID- 11435662 TI - Acute renal allograft rejection: progress in understanding cellular and molecular mechanisms. AB - The application of molecular biology tools to investigate the molecular basis of acute allograft rejection has unravelled many complex mechanisms and improved immunosuppressive therapies leading to significant improvements in graft survival. The "indirect" pathway of antigen presentation has emerged as more important, than the traditional "direct" pathway, for allorecognition by T cells. The recognition that CD28 costimulation is essential for allorecognition has provided novel targets for immunotherapy such as CTLA-Immunoglobulin. Understanding the role of Th1 and Th2 subsets of T helper cells, the cytokine network and cell adhesion molecules in the mediation or prevention of graft rejection has opened new avenues for research into therapeutic modalities. The ideal objective would be to identify the mechanisms of graft destruction and design specific inhibitors. This review highlights recent advances in the understanding of acute renal allograft rejection which may have future potential for rational design of new immunosuppressive strategies. PMID- 11435663 TI - National Institute of Virology. PMID- 11435664 TI - History and development of forensic science in India. PMID- 11435665 TI - Clopidogrel in cardiovascular disorders. PMID- 11435666 TI - Air ambulance services in India. PMID- 11435667 TI - Acoustic nuclei in the medulla and midbrain of the vocalizing Gulf toadfish (Opsanus beta). AB - The organization of the descending and secondary octaval nuclei in the hindbrain of the Gulf toadfish, Opsanus beta, was revealed following the injection of biotin compounds into a physiologically identified auditory region of the torus semicircularis. The results show retrogradely-filled neurons mainly in a dorsomedial division of the descending octaval nucleus, and dorsal and ventral divisions of a secondary octaval nucleus; minor labeling also appeared in dorsolateral and rostromedial intermediate divisions of the descending nucleus. The pattern identified is consistent with that reported in other teleosts, including both vocal and non-vocal species, and clarifies earlier reports of the organization of hindbrain octaval nuclei in toadfish and the closely related midshipman fish. PMID- 11435668 TI - Visual responses of neurons in the pretectal nucleus lentiformis mesencephali to moving patterns within and beyond receptive fields in pigeons. AB - Large-field patterns are effective stimuli for eliciting visual responses from neurons in the pretectal nucleus lentiformis mesencephali of nonmammals. The present study shows that stimulation beyond the receptive field does not contribute to the visual responses of neurons in the nucleus lentiformis mesencephali in two respects. First, changes in the direction and velocity of motion beyond the receptive field did not affect the visual responses of the pretectal cells to motion within the receptive field. Second, time differences in the onset of stimulation within and outside the receptive field did not influence the visual responses of the pretectal cells to motion in the receptive field, implying that there may be no long-range interaction between the receptive field and its surrounding field. The present study also indicates that the pretectal cells are not only sensitive to the direction and velocity of motion, but also to the size and density of dots in a random-dot pattern moving through the receptive field. Taken together with previous studies, these results suggest that the receptive field of the pretectal cells within the nucleus lentiformis mesencephali is large in size but well defined in boundaries, and that the pretectal cells respond to motion of visual stimuli within but not beyond their receptive fields. PMID- 11435669 TI - A comparative analysis of relative brain size in waterfowl (Anseriformes). AB - Variation in relative brain size was examined in 55 species of waterfowl (Anseriformes). Using both conventional statistics and phylogenetically based comparative methods, the extent of variation in relative brain size and possible relationships with mode of foraging and diet were examined. The results indicate that although brain size does vary considerably between closely related species of waterfowl, it is not reliably related to either foraging mode or diet. There are a number of possible reasons for the lack of relationships between brain size and foraging mode and diet. Firstly, subtle changes in foraging mode and diet may favor relatively large changes in brain size. Secondly, foraging mode and diet could be correlated with the expansion of an individual brain region without affecting overall brain size. Thirdly, other behavioral/ecological traits may be more important with respect to brain size evolution in waterfowl. For example, the relatively large brain of the musk duck (Biziura lobata) and altriciality of their young in comparison to other stiff-tailed ducks (Oxyura spp.) indicates that developmental rate plays a significant role in the evolution of brain size. Given the difference between our results and that reported in inter-order comparisons of brain size in birds, further research is required into other avian orders to assess how brain size and behavior might be related within orders as well as between them. PMID- 11435670 TI - Species differences and similarities in the fine structure of the mammalian corpus callosum. AB - A cross-species ultrastructural study of the corpus callosum was performed in six domestic species: the rat, the rabbit, the cat, the dog, the horse and the cow. The results indicate cross-species conservatism in callosal fiber composition with a good interspecies relation between fiber number and brain size. Across species, increases in both brain size and callosal area indicate more callosal fibers, although less than expected from the estimated increase in cortical cell number. Within each species, the correlation between fiber number and brain weight tends to disappear, although in most cases a larger callosum implies a larger number of callosal fibers. The median fiber diameter was conservative across species (0.11-0.2 microm), indicating the maintenance of conduction velocity of most callosal fibers regardless of interhemispheric distance. Nevertheless, the maximal fiber diameters tended to be higher in species with larger brains. Therefore, there is a population of coarse-diameter fibers that tend to increase their diameter and conduction velocity with increasing brain size. However, allometric calculations suggest that the associated increase in velocity in these large fibers may not be sufficient to maintain a constant interhemispheric transmission time in different species. PMID- 11435671 TI - The relationships of brain stem systems to their targets in the spinal cord of the eel, Anguilla anguilla. AB - We have investigated the detail with which supraspinal neurons innervate different regions in the spinal cord. Horseradish peroxidase was applied at different levels of the cord: (1) to the cut surface; (2) injected iontophoretically into one side; or (3) into one of the motoneuron pools innervating different muscle compartments. In all cases, labeled neurons were found throughout the brain distributed over nuclear groups identified in previous fish studies. Some cells from all but one of the nuclei have axons that extend over the whole length of the cord and about 50% of all neurons project to post anal levels. No topographical distribution relating somal position and target location was found. Sixty seven percent of the neurons send axons to the ipsilateral side of the cord, although most nuclear groups provide a bilateral projection. Nucleus ruber projects entirely contralaterally. The descending and magnocellular octavolateral nuclei, and the descending nucleus of the trigeminal nerve project entirely ipsilaterally. Neurons that project to the spinal motoneuron pools innervating the myotomal red- or white-muscle are distributed throughout the same brain stem nuclei, but the cell bodies of those neurons innervating white-muscle motoneurons are larger. Nineteen pairs of cells were found to be consistently identifiable and all project to all levels of the cord; they were only labeled from injections made into the white-muscle motoneuron pool. PMID- 11435672 TI - Circadian variation of stroke onset in Hong Kong Chinese: a hospital-based study. AB - Circadian variation of onset of transient ischaemic attack (TIA) or stroke during four 6-hourly periods starting from midnight was studied in Hong Kong Chinese patients admitted to a regional hospital between October 1996 and July 1999. The onset was classifiable into one of the 6-hourly periods in 832 of 905 patients; patients with unclassifiable onset were more likely to have lacunar infarct and less likely to have intracerebral haemorrhage (ICH). There was a significant circadian variation of onset in all strokes and TIA, TIA alone, ischaemic stroke (IS), ICH and different IS subtypes. The risk of onset was greatest between 6 a.m. and noon for IS or TIA, but between noon and 6 p.m. for ICH. There was no difference in the circadian variation between patients with and without prior TIA or stroke. This hospital-based study revealed a significant circadian variation of onset in different types and subtypes of stroke. PMID- 11435673 TI - Socioeconomic inequalities in the incidence, mortality and prognosis of subarachnoid hemorrhage: the FINMONICA Stroke Register. AB - Low socioeconomic status (SES) is associated with increased mortality from stroke, but usually no distinction is made between stroke subtypes. We analyzed the relationship of SES with mortality and morbidity of subarachnoid hemorrhage (SAH). In the FINMONICA Stroke Register, 956 consecutive SAH events were recorded during 1983-1992 in patients aged 25-74 years. We used taxable income stratified into three categories, low, middle, and high, as an indicator of SES. The age standardized incidence of SAH among men and women aged 25-44 years was approximately three times higher in the low-income group than in the high-income group. In older individuals, differences between the income groups were less pronounced. Among survivors of the acute stage, a poorer prognosis was observed in patients with low income than in those with high income. In conclusion, there is a clear excess mortality and morbidity of SAH in young individuals with low income, particularly among men. PMID- 11435674 TI - Poststroke depression in acute phase after stroke. AB - We studied factors associated with acute poststroke depression in 100 patients, aged 27-70, 2 weeks after their first clinically significant stroke. Depressive symptoms were relatively common (27% Beck Depression Inventory > or =10), but the prevalence of major depression was only 5.6%. Older patients were most vulnerable to poststroke depression. Patients with left hemisphere lesion had no more depression than other patients, but when the lesion was in the left hemisphere or in the brainstem, stroke severity was associated with depression. PMID- 11435675 TI - Suicidal ideas in stroke patients 3 and 15 months after stroke. AB - Patients with stroke may have an increased risk of suicide. We sought to examine the frequency and clinical correlates of suicidal ideas 3 and 15 months after stroke. The study group comprised 286 of 486 consecutive patients aged 55-85 years who at 3 and 15 months after ischemic stroke completed a detailed medical, neurological and radiological stroke evaluation, structured measures of cognition (Mini Mental State Examination), emotion (Beck Depression Inventory, BDI), disability (Rankin scale), and assessment of dependent living. Suicidal ideas were present in 9.8% (n = 28) at 3 months' follow-up and in 14.0% (n = 40) at 15 months' follow-up. The patients with suicidal ideas were more depressed measured by BDI both at 3 (17.0 vs. 7.9, p < 0.001) and 15 months (20.5 vs. 8.5, p < 0.001) after stroke compared to the patients without suicidal ideas. Patients with suicidal ideas had more often a history of a stroke already before the index stroke (35 vs. 18.3%, p = 0.0154), right-sided stroke (60.0 vs. 41.9%, p = 0.0323), they were more disabled according to the Rankin scale (2.4 vs. 1.9; p = 0.0035) and more dependent in living (45.0 vs. 27.2%, p = 0.023) 15 months after stroke. An independent correlate of suicidal ideas 15 months after stroke was history of prior stroke (OR 2.4; 95% CI 1.14-4.97) in logistic regression analysis. The frequency of suicidal ideas increased with the time elapsed from stroke. Recurrent strokes, depressive symptoms, more disabling stroke and right sided stroke correlated to suicidal ideas at 15 months after stroke. Early identification of these risk factors may lead to effective therapeutic intervention. PMID- 11435676 TI - Impact of stroke type on survival and functional health. AB - In a cohort 760 consecutive stroke patients (23 hospitals in the Netherlands), we studied prognosis in relation to stroke type and focused on (a) short-term and long-term mortality, and (b) long-term functional health. Based on clinical and CT data, we distinguished infratentorial strokes from supratentorial strokes (lacunar infarctions, (sub)cortical infarctions and intracerebral hemorrhages). Cumulative mortality for all stroke patients was 34% at 6 months, 51% at 3 years, and 62% at 5 years. Short-term mortality could be explained by stroke type, whereas long-term mortality could not. Of all survivors, 55% were in poor functional health at 6 months, 49% at 3 years and 42% at 5 years. Long-term functional health outcomes were associated with stroke type. We conclude that the impact of stroke type on mortality is limited to the first 6 months, whereas the type of stroke influences the long-term functional health. PMID- 11435677 TI - Brachiofacial pure motor stroke. AB - Pure motor stroke is the commonest lacunar syndrome, but it may be associated with nonlacunar mechanisms of infarction. Pure motor brachiofacial weakness has been considered as a partial syndrome depending on a lacunar mechanism. We studied the correlations between stroke type, topography of infarction and etiology in 22 patients with pure motor brachiofacial weakness who were consecutively admitted to our stroke unit during a 10-year period. Seventeen patients had a small deep infarct, 4 had a cortical infarct in the superficial MCA territory and 1 had no specific lesion. The part of the cardiovascular risk factors was about 36% for smoking, 13% for diabetes mellitus, 60% for dyslipidemia and 40% for heart disease. Hypertension was present in 75% of our cases. None of the patients had a large artery stenosis on Doppler ultrasonography. We concluded that brachiofacial pure motor stroke is not always correlated to lacunar infarcts and may be due to a cortical infarct. MRI should be performed when brain CT is normal because of the implications it may have in management and therapy. PMID- 11435678 TI - Antiepileptic treatment in patients with early postischemic stroke seizures: a retrospective study. AB - Early seizures caused by stroke are a common cause of epilepsy in adults. The protocol for treatment in such a case is not clear. Patients were studied retrospectively after early poststroke seizures. Two groups of patients were compared: one treated group included 35 patients who continued therapy for 2 years; the second group of 23 patients were untreated following a first attack, receiving anticonvulsive therapy only after a second seizure. The data of 61 patients (35 treated and 26 untreated) were analyzed. In the treated group, 24 patients were on treatment with carbamazepine, 9 with valproic acid and 2 with phenytoin. Seizure-free rate after 2 years was 85% in group I and 61% in group II (p = 0.042). Comparing the seizure rate and the event-free period (number/follow up at risk) during the 2 years of no anticonvulsive therapy, both groups experienced about the same seizure rate (4.8 vs. 6.2%, p = 0.605) and similar seizure-free curve survival analysis (p = 0.85). We conclude that treatment of seizures immediately after the first attack after cerebral infarctions decreases the risk of recurrent seizure during the first 2 years of antiepileptic treatment. This subgroup of patients receiving antiepileptic medication immediately after the first poststroke seizure continues to be at the same rate of risk after treatment was stopped as the untreated group during the first 2 years. Antiepileptic therapy is an option in the medical management of poststroke seizure patients, but has no influence on the development of recurrent seizures after discontinuing medication. PMID- 11435679 TI - Prevalence of and risk factors associated with carotid artery stenosis: the Tromso Study. AB - BACKGROUND AND PURPOSE: To assess prevalence, distribution, ultrasound characteristics and determinants of carotid artery stenosis in a large, population-based study of both women and men. METHODS: A total of 6,727 persons aged 25-84 years were screened for extracranial stenosis with Duplex ultrasound of the right carotid artery. Risk factors were compared in 225 persons with stenosis and 5,514 persons without. RESULTS: The prevalence of carotid stenosis was higher in men than in women, where 3.8% (95% CI, 3.2-4.6%) had carotid stenosis, compared to 2.7% (95% CI, 2.2-3.3%) in women (p = 0.001). The prevalence gradually increased by age in both genders. Cholesterol, HDL cholesterol, fibrinogen, systolic blood pressure levels and current smoking were independently associated with carotid artery stenosis in both women and men. The presence of carotid stenosis was significantly associated with a history of cerebrovascular disease, coronary heart disease and peripheral artery disease. For each 10% increase in the degree of carotid stenosis, the risk of having had a cerebrovascular event increased by 26%. CONCLUSIONS: The prevalence of carotid stenosis in the general population, as measured by ultrasound, is low. Age, male gender, smoking, total cholesterol, HDL cholesterol (inverse), fibrinogen and systolic blood pressure are all independent predictors of carotid artery stenosis. PMID- 11435680 TI - Vascular dementia: a cognitive SPET-CBF activation study. AB - OBJECTIVE: We investigated the pattern of regional cerebral blood flow (rCBF) responses to a cognitive task in vascular patients with and without dementia. METHOD: We studied 8 controls and 18 vascular patients by quantitative rCBF assessed by (133)Xe inhalation method and SPET, both at rest and during a cognitive figure recognition task. Eight were mildly demented and 10 were nondemented vascular patients. According to their task performance, 12 patients were classified as 'good performers' (GPs) and 6 patients as 'poor performers' (PPs). RESULTS: Vascular patients activated a larger number of brain areas than controls. No differences were observed between controls, nondemented and mildly demented patients in the pattern of rCBF activation. GPs presented a lower mean percentage of rCBF increase than either controls or PPs. GPs had lower values than PPs in the left temporal, parietal and occipital regions and in the right posterior cingulate and occipital regions. CONCLUSIONS: These data suggest that vascular patients may functionally compensate for vascular damage by activating more brain areas than controls do and, consequently, by increasing the rate of regional activation. PMID- 11435681 TI - Transcranial Doppler detection of cerebral microemboli during left heart catheterization. AB - The role of transcranial Doppler ultrasonography (TCD) in individual risk assessment of embolic complications and the development of prevention strategies during coronary angiography remains to be determined. The purpose of this study was to assess the prevalence, time of occurrence and potential significance of microembolic signals (MES) detected with TCD during femoral left heart catheterization. TCD monitoring of the right and left middle cerebral artery was performed in 51 consecutive patients (36 men, 15 women) who were referred for coronary angiography. Percutaneous coronary angioplasty was performed during the same procedure in 16 patients. MES were counted manually during and after (off line analysis) the procedure. Two patients were excluded from analysis because of the absence of an adequate acoustic temporal window. No neurological event occurred within 24 h in the 49 included patients. MES were detected in all except 2 patients (mean number 17.1 +/- 12.8 per patient), mainly during left ventriculography (38%) and contrast media injection into the coronary arteries (55%), suggesting their gaseous origin. There was no statistically significant association between the number of MES and patient age, cardiovascular history and risk factors, or catheterization results. The presence of coronary artery disease was inversely related to the number of MES (15.8 +/- 0.3 compared to 21.8 +/- 0.2 per patient when a normal angiogram was present; p < 0.05). In conclusion, although asymptomatic microemboli commonly occur during left heart catheterization, the majority of them are probably of gaseous origin, since they occurred predominantly during contrast media injection in this study, and were not related to cardiovascular history or to atheroma risk factors. Because air embolism has been reported to be harmful, attempts to reduce its occurrence during catheter-based procedures could be pertinent. PMID- 11435682 TI - Patient positioning influences oxygen saturation in the acute phase of stroke. AB - We evaluated arterial oxygen saturation (SaO(2)) and heart rate in acute stroke patients to determine whether routine positioning affected these physiological parameters. Measurements were recorded at the bedside non-invasively in five different positions assigned in random order each maintained for 10 min. One hundred and twenty-nine patients examined within a median of 72 h, lying on the left side resulted in slightly lower SaO(2) than lying on the right side, which was statistically significant in the patients with a right (n = 66), but not left, hemiparesis. Patients able to sit in a chair (n = 65), who mostly had less severe strokes, had a significantly higher mean SaO(2) and heart rate when sitting in the chair than when placed in any other position. About 10% of patients, especially those with a severe stroke, with right hemiparesis and concomitant chest disease, experienced falls in SaO(2) to 90% or less for >/=2 min in certain positions; the hypoxia was more likely when they were lying on their left side. These results may have implications for current practice and for future patient positioning strategies to improve outcome after stroke. PMID- 11435683 TI - Microsatellite instability. AB - Unlike aneuploidy, considered to be the cardinal feature of malignant tumors ever since the chromosomal analysis of neoplastic cells became technically feasible, a second pathway toward malignancy has emerged over the past decade that is not characterized by gross aneuploidy but, instead, by inactivation of the DNA mismatch repair system, leading to a hypermutable state in which simple repetitive DNA sequences are unstable during DNA replication. Although mutations of many of these microsatellite sequences are presumably innocuous, because they do not occur in the coding or regulatory regions of genes, other such sequences are critically located in the coding regions of genes involved in the regulation of cell growth. First discovered in the rather uncommon hereditary nonpolyposis colorectal cancer syndrome, where there is an inactivating germline mutation in one of the DNA mismatch repair genes and most of the tumors show microsatellite instability, the latter phenomenon has since been implicated in about 15% of sporadic colorectal cancers, as well as in cancers at several other sites, such as the endometrium. Tumors showing microsatellite instability are generally near diploid, are at a low stage of development, have a favorable prognosis, and, in the colon, are commonly located on the right side. In recent years, epigenetic phenomena, including hypermethylation and loss of imprinting, have come to be recognized as having a significant bearing on the development of these tumors. PMID- 11435684 TI - Expansion of repetitive DNA into cytogenetically visible elements. AB - Two cases of amplified repetitive elements accidentally identified in cancer samples are reported. In both cases, repeated DNA that is normally not visible by traditional chromosome banding had increased in amount to become cytogenetically visible. In one case, an addition to the short arm of chromosome 1 was originally diagnosed. However, upon molecular analysis the diagnosis could be corrected to an amplification of the D1Z2 repeat. In the second case, a strongly DAPI-positive band was visible at the top of the short arm of chromosome 22, and the original diagnosis was add(22). Staining for telomeric repeats revealed their presence inside the DAPI-positive element, thus confirming that the element in question was truly added to the end of the chromosome. Curiously, no telomeric repeats could be detected distal to the DAPI-positive element. The identity of the DAPI positive element could not be established, as it was not stained by any of the specific probes applied, nor in a scanning hybridization with labeled Cot-1 DNA. It thus seems to represent an expansion from some lowly repetitive AT-rich DNA translocated to the tip of chromosome 22. PMID- 11435685 TI - High frequency of spontaneous translocations revealed by FISH in cells from patients with the cancer-prone syndromes ataxia telangiectasia and Nijmegen breakage syndrome. AB - The application of fluorescence in situ hybridization (FISH) using whole chromosome paints (WCPs) is proving to be a very powerful technique for revealing chromosomal instability that, for the most part, has gone undetected by conventional cytogenetic analysis. We have analyzed the frequency of translocations in lymphocytes and lymphoblastoid cell lines from ataxia telangiectasia (AT) and Nijmegen breakage syndrome (NBS) homozygotes and heterozygotes using a three-color chromosome-painting technique (WCP 1, 2, 4). With this assay we were able to detect an increased frequency of spontaneous translocations in AT homozygotes (median, 18.47 +/- 10.82 translocations per 1,000 metaphase cells; 10 patients) and AT heterozygotes (median, 7.87 +/- 3.15 translocations per 1,000 cells; 7 patients), in comparison to controls (median, 2.26 +/- 1.75 translocations per 1,000 cells; 10 controls). Analysis of NBS homozygotes (median, 19.05 +/- 11.27 translocations per 1,000 cells; 5 patients) and NBS heterozygotes (median, 6.93 +/- 3.04 translocations per 1,000 cells; 6 patients) also showed an increased frequency of translocations in these patients compared to controls. The presence of such hitherto undetected chromosomal aberrations corroborate previous findings of spontaneous chromosomal instability in AT and NBS patients, as manifested by an increased rate of open breaks and rearrangements involving chromosomes 7 and 14. Moreover, we show that the degree of genomic instability in AT and NBS patients is even higher than previously established and that some AT and NBS heterozygotes evidence spontaneous chromosomal instability as well. These increased levels of nonspecific translocations could be an important risk factor for the development of malignancies in homozygotes and heterozygotes for ATM or NBS1 gene mutations. PMID- 11435686 TI - Concomitant DNA copy number amplification at 17q and 22q in dermatofibrosarcoma protuberans. AB - Dermatofibrosarcoma protuberans (DFSP) is a tumor of low or intermediate malignant potential with a tendency for recurrence, but low rate of metastasis. The tumorigenesis of DFSP has recently been shown to be associated with the fusion of the collagen type I alpha 1 (COL1A1) and platelet-derived growth factor B-chain (PDGFB) genes, often as a consequence of translocation t(17;22)(q22;q13). Cytogenetically, DFSP is often characterized by supernumerary ring chromosomes containing material from chromosomes 17 and 22. A subset of DFSPs undergo fibrosarcomatous transformation de novo or upon recurrence, and contain components indistinguishable from fibrosarcoma (FS-DFSP). The fibrosarcomatous transformation appears to carry an increased risk for recurrence and metastasis, and is considered to represent tumor progression. The molecular cytogenetic events contributing to tumor progression are unknown. We used comparative genomic hybridization to analyze DNA copy number changes in 11 cases of typical DFSP and 10 cases of FS-DFSP. All cases in both groups were found to exhibit a gain or high-level amplification on chromosome 17q and the majority also on 22q. This finding is in line with previous studies, and suggests further that not only the COL1A1/PDGFB fusion gene formation but also the role of DNA copy number gains in the 17q and 22q regions is crucial per se in the pathogenesis of DFSP. Even though FS-DFSPs displayed a trend toward increase in the number of DNA copy number changes, the difference was not statistically significant, which indicates that mechanisms other than copy number changes are important in the transformation process of DFSP. PMID- 11435687 TI - A novel member of the STOMATIN/EPB72/mec-2 family, stomatin-like 2 (STOML2), is ubiquitously expressed and localizes to HSA chromosome 9p13.1. AB - A cDNA encoding a novel second member of the Band7/stomatin-like/SPFH domain family in humans designated stomatin-like 2 (STOML2) has been isolated using the technique of cDNA Representational Difference Analysis. The STOML2 cDNA encoded a 356 amino acid residue polypeptide with a predicted molecular weight of 38.5 kDa. The predicted polypeptide sequence of STOML2 could be delineated into three major domains: an N-terminal alpha-helical region; a domain with significant similarity to a 172 amino acid region of the HSA stomatin polypeptide, composed of an alternating alpha-helical and beta-sheet structure and a C-terminal domain that was mostly alpha-helical. The stomatin-like domain was observed in 51 other proteins with potentially diverse functions. Based on its homology to stomatin, STOML2 was predicted to be cytoplasmically located. However, unlike most of the other proteins containing stomatin-like domains, the predicted STOML2 polypeptide does not contain a transmembrane region although the presence of N-myristoylation sites suggest that it has the potential to be membrane-associated. Northern blot analysis of a panel of poly(A)(+) mRNA from normal human adult tissues showed that a single 1.3-kb mRNA transcript encoding STOML2 was ubiquitously expressed, with relatively higher levels in skeletal muscle and heart compared to other tissues. Comparison of the STOML2 cDNA sequence with human genomic DNA indicated that the gene encoding STOML2 was 3,250 bp long and consisted of ten exons interrupted by nine introns. We have mapped STOML2 to HSA chromosome 9p13.1, a region that is rearranged in some cancers and thought to contain the gene responsible for acromesomelic dysplasia. PMID- 11435688 TI - An ATP-binding cassette gene (ABCG5) from the ABCG (White) gene subfamily maps to human chromosome 2p21 in the region of the Sitosterolemia locus. AB - We characterized a new human ATP-binding cassette (ABC) transporter gene that is highly expressed in the liver. The gene, ABCG5, contains 13 exons and encodes a 651 amino acid protein. The predicted protein is closely related to the Drosophila white gene and a human gene, ABCG1, which is induced by cholesterol. This subfamily of genes all have a single ATP-binding domain at the N-terminus and a single C-terminal set of transmembrane segments. ABCG5 maps to human chromosome 2p21, between the markers D2S117 and D2S119. The abundant expression of this gene in the liver suggests that the protein product has an important role in transport of specific molecule(s) into or out of this tissue. PMID- 11435689 TI - Refined localization of twenty-one genes in subregion p13.1 of human chromosome 1. AB - In this report, we describe a refinement of the human transcript map of chromosome 1p13.1, a subregion undergoing many aberrations in various types of human cancers. Publicly available genetic linkage, radiation hybrid and physical maps, as well as cytogenetic and sequence data were used to establish the relative order and orientation of ten known intragenic markers. The complete sequence of genomic clones of the region, available at the Sanger Centre, provided the tool for further studies performed by BLAST analysis against all cDNA sequences registered in the Genexpress Index2. This allowed us to assign to subband 1p13.1 nine of the ten known genes, an additional member of the gene family of one of these genes and eleven new transcripts. The remaining known gene and one additional new transcript map at the 1p13.1 and 1p13.2 boundary. The corresponding genes may be responsible for disorders related to this region. The resulting transcript map of 1p13.1 is presented in the printed article with additional data available on a dedicated Web site at the address http://idefix.upr420.vjf.cnrs.fr/CARTO. PMID- 11435690 TI - Gene for the human transmembrane-type protein tyrosine phosphatase H (PTPRH): genomic structure, fine-mapping and its exclusion as a candidate for Peutz Jeghers syndrome. AB - Mutations in the serine/threonine kinase STK11 lead to Peutz-Jeghers syndrome (PJS) in a subset of affected individuals. Significant evidence for linkage to a second potential PJS disease locus on 19q13.4 has previously been described in one PJS family (PJS07). In the current study, we investigated this second locus for PJS gene candidates. We mapped the main candidate gene in this region, the gene for the transmembrane-type protein tyrosine phosphatase H (PTPRH), within 15 kb telomeric to the marker D19S880. We determined its genomic structure, and performed mutation analysis of all exons and the exon-intron junctions of the PTPRH gene in the PJS07 family. No disease causing mutation was identified in PTPRH in affected individuals, suggesting the existence of an as yet not identified gene on 19q13.4 as a second PJS gene. PMID- 11435691 TI - Genomic organization and refined mapping of the human nuclear corepressor 2 (NCOR2)/ silencing mediator of retinoid and thyroid hormone receptor (SMRT) gene on chromosome 12q24.3. AB - The human nuclear co-repressor 2 (N-CoR2) gene (NCOR2, previously called silencing mediator for retinoid and thyroid hormone receptor SMRT) is recruited to nuclear and non-nuclear receptors in a large repressing complex containing also N-CoR1, mSin3 and HDACs. This large complex represses transcription in absence of ligand. Herein we report the high- resolution and refined mapping of NCOR2 at the boundary of sub-bands 12q24.23 and 12q24.31, and its intron/exon structure. The gene contains 45 exons. This information should allow further study of potential NCOR2 genomic alteration in some subsets of malignancies. PMID- 11435692 TI - Human puromycin-sensitive aminopeptidase: cloning of 3' UTR, evidence for a polymorphism at a.a. 140 and refined chromosomal localization to 17q21. AB - Puromycin-sensitive aminopeptidase is a predominantly cytoplasmatic zinc dependent exopeptidase. Its physiological function is not known to date. Here we report data on tissue distribution, a polymorphism within the coding region and the complete 3' UTR. The gene (NPEPPS alias PSA) was physically mapped to chromosome 17q21.2-->q21.32 using fluorescence in situ hybridization. PMID- 11435693 TI - cDNA cloning, chromosome assignment, and genomic structure of a human gene encoding a novel member of the RBM family. AB - We have cloned and characterised a novel human gene mapping to chromosome 20q11.2. A partial transcript was initially isolated from a human cDNA library transcribed from RNA of the colon carcinoma cell line T-84. In order to determine the full coding sequence of this novel mRNA, we isolated seven cDNA clones from a human cDNA library transcribed from RNA of the acute monocytic leukemia cell line THP1 by colony hybridization. On Northern blot analysis of four human cell lines, the cDNAs isolated hybridize with an abundantly expressed mRNA species of 3.5 kb. A full-length cDNA transcript of this novel mRNA has an open reading frame of 2,796 bp encoding a protein with a calculated molecular weight of 97 kDa. Two repetitive structural consensus motifs are contained within the deduced protein sequence, namely five distinct RNA binding motifs and two proline rich regions. The derived protein sequence also contains putative transmembrane domains. These structural motifs identify this novel protein as a member of an expanding protein family containing RNA binding motifs (RBM). As seen from recently completed sequence of the genomic area encoding this novel mRNA by the Sanger Centre Human Genome Project, the coding region of this gene, RBM12, is intronless. PMID- 11435694 TI - Molecular cloning and characterization of a novel peptidylprolyl isomerase (cyclophilin)-like gene (PPIL3) from human fetal brain. AB - During the large-scale sequencing analysis of a human fetal brain cDNA library, we isolated two cDNA clones encoding two novel proteins, which show 52% and 72% identity to the cyclophilin isoform 10 of C. elegans, respectively. Sequence analysis revealed these two cDNA clones are two different splicing variants of a novel cyclophilin-like gene (PPIL3). The PPIL3 gene was identified on a completely sequenced BAC (GenBank accession AC005037) from chromosome 2q33 between STS markers stSG2762 (proximal) and SHGC-3074 (distal), oriented toward the telomere. The PPIL3 gene consisted of eight exons spanning more than 18 kb of genomic DNA. RT-PCR analysis indicated that PPIL3 was ubiquitously expressed in adult human tissues. PMID- 11435695 TI - Polymorphism and polytypy for pericentric inversions in 38-chromosome Mastomys (Rodentia, Murinae) and possible taxonomic implications. AB - Chromosome banding analysis (R- and C-bands) of two 38-chromosome Mastomys specimens originating from the Ivory Coast and Uganda revealed different numbers of autosome arms (NFa), equal to 51 and 60, respectively. Comparison of their chromosome banding patterns with those of Mastomys specimens from the Sudan (NFa = 41) and Senegal (NFa = 51-54), studied previously, showed that variation of the NFa from 40 to 60 throughout the species distribution is the result of a pericentric inversion polymorphism involving 3-12 chromosome pairs. At the population level, this variation is much narrower and never results from more than two chromosome pairs involved in inversion polymorphism. Taking into account that the NFa values recorded to date form a well-defined discontinuous row, we presume that introgressive hybridization between populations differing from each other by 3-5 to 11-12 pericentric inversions is interrupted. From there, the hypothesis of the existence of at least three cryptic species (designated provisionally as MER-1, MER-2, and MER-3) within 38-chromosome Mastomys populations previously assigned to M. erythroleucus can be made. It looks likely that one of them, possessing a karyotype with an NFa = 50-56, is widely distributed throughout sub-Saharan Africa and includes karyotyped populations from Senegal, the Ivory Coast, Mali, Benin, Cameroon, Zaire, and the Sudan. The second species (MER-2) includes the specimens karyotyped (NFa = 40-41) from Chad and the Sudan. Finally, a third tentative species (MER-3) corresponds to specimens with NFa = 59-60 found in East Zaire and Uganda, as well as possibly Mali and Chad. PMID- 11435696 TI - Phylogenetic implications of the 38 putative ancestral chromosome segments for four canid species. AB - Chromosome homologies between the Japanese raccoon dog (Nectereutes procyonoides viverrinus, 2n = 39 + 2-4 B chromosomes) and domestic dog (Canis familiaris, 2n = 78) have been established by hybridizing a complete set of canine paint probes onto high-resolution G-banded chromosomes of the raccoon dog. Dog chromosomes 1, 13, and 19 each correspond to two raccoon dog chromosome segments, while the remaining 35 dog autosomes each correspond to a single segment. In total, 38 dog autosome paints revealed 41 conserved segments in the raccoon dog. The use of dog painting probes has enabled integration of the raccoon dog chromosomes into the previously established comparative map for the domestic dog, Arctic fox (Alopex lagopus), and red fox (Vulpes vulpes). Extensive chromosome arm homologies were found among chromosomes of the red fox, Arctic fox, and raccoon dog. Contradicting previous findings, our results show that the raccoon dog does not share a single biarmed autosome in common with the Arctic fox, red fox, or domestic cat. Comparative analysis of the distribution patterns of conserved chromosome segments revealed by dog paints in the genomes of the canids, cats, and human reveals 38 ancestral autosome segments. These segments could represent the ancestral chromosome arms in the karyotype of the most recent ancestor of the Canidae family, which we suggest could have had a low diploid number, based on comparisons with outgroup species. PMID- 11435697 TI - Conserved chromosome segments in Hylobates hoolock revealed by human and H. leucogenys paint probes. AB - A complete comparative chromosome map of the white-browed gibbon (Hylobates hoolock, 2n = 38), white-cheeked gibbon (Hylobates leucogenys, 2n = 52), and human has been established by hybridising H. leucogenys chromosome-specific paints and human 24-colour paints onto H. hoolock metaphase chromosomes. In the 18 H. hoolock autosomes, we identified 62 conserved segments that showed DNA homology to regions of the 25 H. leucogenys autosomes. Numerous interchromosomal rearrangements differentiate the karyotypes of H. leucogenys and H. hoolock. Only H. hoolock chromosome 10 showed homology to one entire autosome of H. leucogenys. The hybridisation of human 24-colour paints not only confirmed most of the chromosome correspondences between human and H. hoolock established previously but also helped to correct five erroneous assignments and revealed three new segments. Our results demonstrate that the karyotypes of the extant gibbons have arisen mainly through extensive translocation events and that the karyotype of H. hoolock more closely resembles the ancestral karyotype of Hylobates, rather than the karyotype of H. leucogenys. PMID- 11435698 TI - Molecular cytogenetic analysis of the highly repetitive DNA in the genome of Apodemus argenteus, with comments on the phylogenetic relationships in the genus Apodemus. AB - The DNA of Apodemus argenteus was digested with DraI, and the resultant DraI fragment of highly repetitive DNA was isolated and analyzed by DNA filter hybridization, cloning, sequencing, and fluorescence in situ hybridization (FISH). Southern blot hybridization and nucleotide sequencing revealed that most of the DraI fragment consisted of a 230-bp repeating unit and contained no sex chromosome-specific nucleotide sequences. The DraI fragment included the CENP-B box-like sequence, with a strong homology to the human CENP-B box sequence. FISH revealed that the DraI fragment was specific to all pericentromeric C-band positive regions, as well as to the C-block of the X chromosome. No hybridization signals were obtained from A. speciosus, A. peninsulae peninsulae, A.p. giliacus, A. agrarius, A. sylvaticus, A. semotus, or Mus musculus when the DraI fragment was used as probe. Peptide nucleic acid (PNA)-FISH using the CENP-B box-like sequence in the DraI fragments as probe suggested that this nucleotide sequence was also specific to all pericentromeric C-heterochromatic regions of A. argenteus chromosomes. Zoo-blot hybridization using DraI-digested genomic DNA from three species of Apodemus (namely, A. argenteus, A. speciosus, and A. peninsulae) and from Mus musculus strongly suggested that the consensus DraI fragment contained nucleotide sequences that were species-specific for A. argenteus. These results also suggest that A. argenteus is phylogenetically distant from other Apodemus species examined, as well as the possibility that the DraI fragment might be related directly to the delayed quinacrine mustard fluorescence of many pericentromeric C-heterochromatic regions of the chromosomes in A. argenteus. PMID- 11435699 TI - Comparative analysis of the promoter structure and genomic organization of the human and mouse ABCA7 gene encoding a novel ABCA transporter. AB - We report here the genomic and transcriptional characterization in mouse and man of a novel transporter of the ABCA subclass, named ABCA7. As it is the case for other ABCA genes, the predicted protein encoded by ABCA7 is a full symmetric transporter, highly conserved across species. The ABCA7 gene maps to human chromosome 19 and to the homologous region at band B4-C1 on mouse chromosome 10. The preferential expression of ABCA7 in the spleen, thymus, and fetal liver is consistent with the finding, in both human and mouse promoter, of sites targeted by lymphomyeloid-specific transcription factors. This suggests that ABCA7 may play a pivotal role in the developmental specification of hematopoietic cell lineages. PMID- 11435700 TI - Structure, sequence and function of a marsupial LIF gene: conservation of IL-6 family cytokines. AB - Leukaemia Inhibitory Factor (LIF) is a multifunctional cytokine with an obligate role in the mouse in embryonic implantation. In this paper we demonstrate the existence of a functional LIF gene in the marsupial Sminthopsis crassicaudata, and the presence of LIF-related sequences in the monotreme Tachyglossus aculeatus (Australian echidna). Isolation of genomic and cDNA clones from S. crassicaudata, indicated that the LIF gene is highly conserved between marsupials and monotremes in terms of sequence and genomic organisation. Critical functional residues within the LIF sequence were also conserved including residues implicated in intracellular LIF activity, and in interaction with the receptor subunits LIFR and gp130. These findings suggest that the structure and biochemical function of the protein is likely to be conserved. Consistent with this, purified recombinant S. crassicaudata LIF interacted functionally with mouse receptor components and was sufficient for maintenance of mouse embryonic stem (ES) cells in the undifferentiated state. Conservation of LIF outside eutherians is intriguing given the markedly divergent reproductive strategies which include, for some marsupial species, embryonic diapause, and in monotremes, the absence of implantation. The availability of marsupial LIF probes provides an opportunity to investigate conservation of expression and function in these mammals. PMID- 11435701 TI - Cloning and molecular characterization of three ubiquitin fusion degradation 1 (Ufd1) ortholog genes from Xenopus laevis, Gallus gallus and Drosophila melanogaster. AB - The yeast ubiquitin fusion degradation 1 (Ufd1) protein is involved in a degradation pathway for ubiquitin fused products. The human ortholog gene (UFD1 like, UFD1L) is deleted in patients affected by the DiGeorge/velocardiofacial syndromes. We report the cloning of UFD1L orthologs from Drosophila melanogaster (dufd1l), Xenopus laevis and Gallus gallus. The 1,125-bp Drosophila cDNA encodes a protein of 316 amino acids, showing 60% identity with the human and murine proteins. The identity to the G. gallus, X. laevis, C. elegans and S. cerevisiae proteins is 95%, 83%, 32%, and 36%, respectively. Northern expression data in Drosophila indicate that dufd1l is expressed through embryonic, larval and pupal development, as well as in the adult fly. PMID- 11435702 TI - International System for Chromosome Nomenclature of Domestic Bovids (ISCNDB 2000). PMID- 11435703 TI - The human hedgehog-interacting protein gene: structure and chromosome mapping to 4q31.21-->q31.3. AB - Hedgehog-interacting protein (Hhip) is a novel regulatory component in the vertebrate hedgehog-signalling pathway. The murine Hhip encodes a type I TM protein that attenuates hedgehog signalling by binding all three mammalian hedgehog proteins. Here we describe the cloning and characterisation of the homologous human hedgehog-interacting protein gene (HHIP). HHIP comprises 13 exons and spans >91kb encoding a protein of 700 aa which shares 94% sequence iden tity with mouse Hhip. HHIP maps to chromosome 4q31.21--> q31.3. Additionally, we have mapped murine Hhip to chromosome 8. PMID- 11435704 TI - Identification and isolation of a full-length clone of mouse GMFB (Gmfb), a putative intracellular kinase regulator, differentially expressed in telencephalon. AB - We identified new transcribed sequences, using a differential display paradigm to select genes expressed in proliferating neuroblasts from mouse telencephalon at 10 days of embryonic development. In this systematic search, we isolated a 361-bp partial 3' untranslated region (3' UTR) homologous to the 3' UTR of the human gene encoding a putative intracellular kinase regulator, glia maturation factor beta (GMFB). We cloned a full-length, 4,311-bp mouse cDNA containing a 270-bp 5' UTR, a 3,615-bp 3' UTR, and an open reading frame of 426 nucleotides encoding a putative 142 amino-acid protein, identical to human GMFB, with the exception of two amino acids. This 4.3-kb transcript is present in a variety of adult tissues and is developmentally regulated as shown by Northern blot analysis. Differential expression in telencephalon was demonstrated by quantification of radioactive relative RT-PCR and confirmed by in situ hybridization. The isolation of this full-length clone of mouse Gmfb should facilitate investigation of the intracellular mechanisms involved in the development of telencephalon. PMID- 11435705 TI - Mapping and characterization of the mouse and human SS18 genes, two human SS18 like genes and a mouse Ss18 pseudogene. AB - We have previously isolated and characterized a mouse cDNA orthologous to the human synovial sarcoma associated SS18 (formerly named SSXT and SYT) cDNA. Here, we report the characterization of the genomic structure of the mouse Ss18 gene. Through in silico methods with sequence information contained in the public databases, we did the same for the human SS18 gene and two human SS18 homologous genes, SS18L1 and SS18L2. In addition, we identified a mouse Ss18 processed pseudogene and mapped it to chromosome 1, band A2-3. The mouse Ss18 gene, which is subject to extensive alternative splicing, is made up of 11 exons, spread out over approximately 45 kb of genomic sequence. The human SS18 gene is also composed of 11 exons with similar intron-exon boundaries, spreading out over about 70 kb of genomic sequence. One alternatively spliced exon, which is not included in the published SS18 cDNA, corresponds to a stretch of sequence which we previously identified in the mouse Ss18 cDNA. The human SS18L1 gene, which is also made up of 11 exons with similar intron-exon boundaries, was mapped to chromosome 20 band q13.3. The smaller SS18L2 gene, which is composed of three exons with similar boundaries as the first three exons of the other three genes, was mapped to chromosome 3 band p21. Through sequence and mutation analyses this gene could be excluded as a candidate gene for 3p21-associated renal cell cancer. In addition, we created a detailed BAC map around the human SS18 gene, placing it unequivocally between the CA-repeat marker AFMc014wf9 and the dihydrofolate reductase pseudogene DHFRP1. The next gene in this map, located distal to SS18, was found to be the TBP associated factor TAFII-105 (TAF2C2). Further analogies between the mouse Ss18 gene, the human SS18 gene and its two homologous genes were found in the putative promoter fragments. All four promoters resemble the promoters of housekeeping genes in that they are TATA-less and embedded in canonical CpG islands, thus explaining the high and widespread expression of the SS18 genes. PMID- 11435706 TI - Cloning and chromosome mapping of human and chicken Iroquois (IRX) genes. AB - Three highly homologous homeobox genes (caupolican, araucan and mirror) have been identified in Drosophila. These genes belong to the novel Iroquois complex, which acts as a pre-pattern molecule in Drosophila neurogenesis. Recently several vertebrate Iroquois homologues (Irx) were isolated and found to be involved in pattern formation of various tissues. Here we report cytogenetic mapping of four human and five chicken Iroquois genes by FISH. Our findings revealed that vertebrate Irx genes are clustered at two different loci. PMID- 11435707 TI - Genomic structure, chromosomal localization, and embryonic expression of the mouse homolog of PRCC, a gene associated with papillary renal cell carcinoma. AB - In a subset of papillary renal cell carcinomas a t(X;1)(p11;q21) chromosome translocation has repeatedly been reported. Positional cloning has demonstrated that, as a result of this translocation, the transcription factor TFE3 gene on the X-chromosome becomes fused to a novel gene, PRCC, on chromosome 1. Since as yet little is known about the function of PRCC, we sought to identify the mouse counterpart of the PRCC gene. Isolation and sequence analysis of a mouse Prcc cDNA revealed a high level of conservation between man and mouse, both at the nucleotide and protein level. As the human PRCC gene, the mouse Prcc gene is ubiquitously expressed. It shows low expression in all mouse fetal tissues examined. In addition, we identified a genomic cosmid clone containing the complete Prcc gene. The mouse Prcc gene consists of seven exons, all of which contain coding sequences. The small second exon, which was found to be located adjacent to the t(X;1) breakpoint in the human gene on chromosome 1, is also conserved between man and mouse. In mouse, Prcc is located on chromosome 3. These cDNA and genomic clones will be instrumental in the creation of mouse models for a further elucidation of the function of PRCC. PMID- 11435708 TI - Structure and chromosome location of the mouse P2X(1) purinoceptor gene (P2rx1). AB - P2X(1) receptors are ATP-gated cation channels that mediate the fast, purinergic component of sympathetic nerve-smooth muscle neurotransmission in the mouse vas deferens and may serve comparable functions in the urinary bladder and the arteries. The gene for mouse P2X(1) (P2rx1) was cloned and its genomic structure defined by sequencing. The gene spans about 10 kb and consists of 12 exons. All splice sites conformed to the GT-AG motif and the exon-intron boundaries were largely conserved with other members of the P2X gene family so far cloned. A single transcription-starting site was identified by 5' RACE analysis, 233 bp upstream of the translation start site. The P2X(1) gene maps to the central region of mouse chromosome 11. PMID- 11435709 TI - cDNA cloning, chromosomal localization and evolutionary analysis of mouse vacuolar ATPase subunit D, Atp6m. AB - The multi-subunit vacuolar ATPase pump uses ATP hydrolysis to move protons into membrane bound compartments. The pump is involved in a variety of cellular functions, including regulation of cytosolic pH, vesicular transport, endocytosis, secretion, and apoptosis. Here, we describe the cDNA cloning and chromosomal mapping of subunit D of murine V-ATPase. The mouse gene, designated Atp6m, maps to Chromosome 12, in a region of high homology with human chromosome 14q24. Evolutionary analysis of subunit D orthologs in a variety of other species reveals that this is a highly conserved protein that has been under remarkably strong negative selection during evolution, most likely reflecting its critical role in multiple cellular processes. PMID- 11435710 TI - Transmission of Robertsonian chromosomes through human female meiosis. PMID- 11435711 TI - Are human cancers ever diploid - or often trisomic?: an update. PMID- 11435712 TI - Assignment of TRADD to human chromosome band 16q22 by in situ hybridization. PMID- 11435713 TI - Isolation and mapping of the pig homolog of the Wiskott-Aldrich Syndrome gene (WAS). PMID- 11435714 TI - Isolation and mapping the pig homologs survivin (BIRC5) and effector cell protease receptor 1 (EPR1) genes. PMID- 11435715 TI - Assignment of BCL2L11 to human chromosome band 2p13 with somatic cell and radiation hybrids. PMID- 11435716 TI - Assignment of the HOX11L2 gene to human chromosome band 5q35.1 and of its murine homolog to mouse chromosome bands 11A4-A5 by in situ hybridization. PMID- 11435717 TI - Assignment of the bovine ectodysplasin A gene (ED1) to bovine Xq22-->q24 by fluorescence in situ hybridization. PMID- 11435718 TI - Assignment of the human homeobox 11-like 2 gene (HOX11L2) to chromosome 5q34- >q35 by radiation hybrid mapping. PMID- 11435719 TI - Effects of lansoprazole on human gastric lipase secretion and intragastric lipolysis in healthy human volunteers. AB - BACKGROUND: Lansoprazole is a potent proton-pump inhibitor (PPI) of parietal cells, which reduces the secretion of gastric acid. Although human gastric lipase (HGL) is produced only by the chief cells of the stomach, the possibility that interactions may occur between lansoprazole and HGL has never been addressed so far in humans. The aim of this study was therefore to quantify the effects of lansoprazole on HGL secretion and intragastric lipolysis during the ingestion of test meals by healthy human volunteers. METHODS: Six healthy volunteers were intubated twice with a gastric and a duodenal tube, before ingesting a standard liquid test meal alone (-PPI experiments) and after 7 days of lansoprazole treatment (+PPI experiments). The HGL concentration was assessed in gastric and duodenal samples by measuring the lipase activity using a pH-stat, and the lipolysis products were quantified by performing thin layer chromatography. The level of intragastric lipolysis was defined as the percentage acyl chains released from the meal triglycerides. The pyloric outputs of HGL and lipolysis products were calculated, based on the use of a non-absorbable marker added to the meal. RESULTS: The pH of the gastric contents was significantly higher in the +PPI experiments than in the -PPI experiments (p < 0.05), since mean values of 4.3 +/- 2.5 and 2.2 +/- 1.6, respectively, were recorded at the end of the gastric emptying of the meal. The HGL concentrations recorded during the meal were found to be higher in the experiments with lansoprazole (p < 0.05) than in those without lansopra- zole, but the HGL secretion levels (-PPI: 15.4 +/- 8.0 mg; +PPI: 19.0 +/- 7.4 mg) and the intragastric lipolysis (-PPI: 24.0 +/- 8.0%; +PPI: 23.6 +/- 6.8%) were not significantly affected by lansoprazole (p > 0.05 in both cases). CONCLUSION: Lansoprazole affected neither the HGL secretion nor the intragastric lipolysis levels, although an increase was observed in the intragastric pH at the end of the gastric emptying of the meal. The HGL concentration increased, however, due to the decrease in the acid secretion process, resulting in less diluted gastric contents. PMID- 11435720 TI - Induction of gastric ornithine decarboxylase in early weaning rats. AB - BACKGROUND/AIMS: Early weaning has been shown to induce intestinal ornithine decarboxylase (ODC) activities and cell proliferation in rats. No information is available about the effect of early weaning on ODC activity in the stomach. METHODS: Suckling rats were prematurely weaned on postnatal day 15 and followed through day 21. Oxyntic gland mucosa of stomach was obtained on postnatal days 15, 16, 18 and 21 (days 0, 1, 3 and 6 after early weaning) and assayed for ODC activity, DNA, protein and pepsinogen activity. alpha-Difluoromethyl ornithine (DFMO), a specific ODC inhibitor, was given orally to early-weaned pups and its resultant effects were assessed on days 1 and 6 after early weaning. RESULTS: Stomach mucosal wet weight, DNA, protein and pepsinogen activities significantly increased on day 6 after early weaning. ODC activity increased on days 1, 3, and 6 after early weaning, with the highest increase (3-fold) on day 1 when compared to controls. The increases of ODC activity, DNA and protein contents as induced by early weaning were significantly suppressed when pups were exposed to DFMO. However, no suppression of pepsinogen activity was observed. CONCLUSIONS: Our study shows that early weaning induces ODC activity and functional growth in the stomach. Gastric ODC activity is essential in gastric mucosal growth processes but not in differentiation. The induction of stomach ODC may act as an early marker in the growth of stomach mucosa induced by early weaning in rats. PMID- 11435721 TI - Factor XIII improves gastric stress lesions in rats. AB - BACKGROUND/AIMS: Tissue transglutaminase has been reported to be involved in the healing of experimental gastric ulcer; nevertheless, other type(s) of transglutaminase could be involved. The present experiments aimed at examining whether plasma transglutaminase (factor XIIIa) contributes to such healing and at evaluating whether factor XIII supplementation improves gastric mucosal lesions. METHODS: The healing effect of 200 U/kg of factor XIII administered intravenously was examined using a water immersion restraint rat model of stress gastric damage. The rats were sacrified 0, 2, 4, and 12 h after stress. The gastric mucosa was examined macroscopically and microscopically, and the transglutaminase activities were assayed in serum and gastric mucosa. Factor XIIIa and tissue transglutaminase protein levels in the gastric mucosa were analyzed by immunoblot. Immunohistochemistry was used to identify the location of tissue transglutaminase, factor XIIIa, and fibronectin in the gastric mucosa. RESULTS: The transglutaminase activity, reduced by stress in the gastric mucosa, increased up to 12 h after stress, peaking at 4 h, when the ulcer index significantly decreased. The serum transglutaminase level was low at all time points. Exogenous administration of factor XIII allowed a faster reduction of the ulcer index that was coincident with an increased transglutaminase activity in the mucosa. Both tissue transglutaminase and factor XIIIa protein levels were reduced by 6 h of stress and increased after factor XIII administration. Immunohistochemistry showed a colocalization of both factor XIIIa and tissue transglutaminase with fibronectin in the extracellular matrix of the damaged area. CONCLUSIONS: Two forms of transglutaminase are involved in the healing of stress-induced gastric erosions, and factor XIII administration allows faster gastric mucosa healing. PMID- 11435722 TI - K-ras and p53 mutations in colonic lavage fluid of patients with colorectal neoplasias. AB - BACKGROUND: The adenoma-carcinoma sequence has its molecular basis in several gene mutations of which K-ras and p53 are of paramount importance. The aims of this study were to evaluate whether these genetic alterations can be detected in colonic lavage fluid from patients with colorectal adenomas and carcinomas. METHODS: In 45 patients with adenomas, 20 patients with colorectal carcinomas and 38 patients with non-neoplastic and noninflammatory diseases of the colon p53 and K-ras mutations were evaluated in colonic lavage fluid employing single-strand confirmation polymorphism analysis and dot-blot hybridization, respectively. RESULTS: Mutations of the K-ras and the p53 gene were found in 15.6% (p = 0.065) of patients with adenomas, in 25.0 % (p = 0.016) of patients with carcinomas and in 2.6% in the control group. CONCLUSION: Genetic alterations in the colonic lavage fluid could be an additional diagnostic tool for the surveillance of patients with colorectal neoplasias. PMID- 11435723 TI - Beneficial effects of Batimastat (BB-94), a matrix metalloproteinase inhibitor, in rat experimental colitis. AB - BACKGROUND AND AIMS: Matrix metalloproteinases (MMPs) represent a group of enzymes that regulate cell-matrix composition playing a major role in the inflammatory response. In the present study we evaluated the ability of the MMP inhibitor Batimastat (BB-94) to modify the course of experimental colitis induced in the rat by trinitrobenzensulfonic acid (TNB). METHODS: Colitis was induced in 40 rats by intracolonic administration of TNB. Animals were divided into four groups of ten rats each: group 1 received only intracolonic TNB, group 2 received TNB+5 mg/kg intraperitoneal BB-94, group 3 TNB+10 mg/kg BB-94 and group 4 TNB+20 mg/kg BB-94. The MMP inhibitor was administered 30 min before induction of colitis and twice daily until death. Ten rats receiving only intracolonic 0.9% saline served as controls. Animals were killed after seven days; segments of colon were removed and used for histological score of inflammation and myeloperoxidase (MPO) activity. RESULTS: Rats receiving only intracolonic 0.9% saline showed no evidence of colitis. The inflammation score was 0.9, MPO activity 0.235 U/mg. Group 1 (TNB-treated rats) exhibited a high inflammation score (12.4) and MPO activity (0.715 U/mg). Conversely, BB-94-treated rats showed, compared to the TNB group, a significantly lower inflammation score and MPO activity in a dose-dependent fashion. Group 2: inflammatory score 10.1, MPO activity 0.474 (p < 0.05 vs. TNB); group 3: inflammatory score 8.3, MPO activity 0.287 (p < 0.01 vs. TNB); group 4: inflammatory score 5.0, MPO activity 0.256 (p < 0.01 vs. TNB). CONCLUSIONS: Treatment with BB-94 has dose-dependent beneficial effects on the inflammatory alterations in rat experimental colitis. Thus, the inhibition of MMPs may represent a novel therapeutic approach for treatment of intestinal inflammation. PMID- 11435724 TI - Quality of life of Greek patients with inflammatory bowel disease. Validation of the Greek translation of the inflammatory bowel disease questionnaire. AB - BACKGROUND/AIMS: Health-related quality of life (HRQOL) is an important outcome measure in inflammatory bowel disease (IBD). The aim of our study is to validate the Greek translation of the Inflammatory Bowel Disease Questionnaire (IBDQ). For this we assessed its construct validity, discriminant ability, reliability and sensitivity to change. METHODS: One hundred and fourteen patients with IBD (69 UC, 45 CD) completed the Greek version of the IBDQ, and a Visual Analogue Scale (VAS) for general well-being. Fifty-two patients also completed the SF-36. A subgroup of 46 patients completed the Greek IBDQ for a second time. Clinical activity was assessed by Harvey-Bradshaw Index and Colitis Activity Index. RESULTS: Correlation coefficients between the four dimensional scores of the Greek IBDQ and the clinical activity indexes, VAS and the SF-36 were all reasonably high and statistically significant. The Greek IBDQ was able to discriminate well between groups of patients with either different clinical disease activity or subjective assessment of well-being. It also showed high reliability when it was repeated in patients who reported no change in their general well-being (intraclass correlation coefficient 0.940-0.998). In contrast, there was a significant difference between the baseline and the follow-up measurement in patients who reported change in their general well-being. CONCLUSIONS: The Greek IBDQ proved to be a valid and reliable instrument for assessing HRQOL, useful in the evaluation of clinical trials or health surveys as well as in the therapeutic management of IBD patients. PMID- 11435725 TI - Are mast cells MASTers in HIV-1 infection? AB - HIV-1 gp120 interacts with IgE V(H)3(+) on the surface of human basophils and mast cells (Fc epsilon RI(+) cells), acting as a viral immunoglobulin superantigen. gp120 from different clades induces mediator release from Fc epsilon RI(+) cells. gp120 also induces IL-4 and IL-13 synthesis in human basophils. The chemokine receptors CCR3 and CXCR4, which are coreceptors of HIV-1 infection, are expressed by human Fc epsilon RI(+) cells. HIV-1 Tat protein is a potent chemoattractant for basophils and lung mast cells, interacting with CCR3. Incubation of basophils with Tat protein upregulates the surface expression of the CCR3 receptor. There is evidence that human Fc epsilon RI(+) cells could be infected in vitro by M-tropic HIV-1 strains. PMID- 11435726 TI - Recombinant allergens for skin testing. AB - Skin testing is a basic diagnostic procedure widely used to explore immediate type reactions to allergen preparations in vivo. Despite their reliability, if standardized extracts are used, skin tests suffer from limited reproducibility due to difficulties in preparing consistently standardized extracts from natural raw material. Starting from allergen-encoding cDNAs, large amounts of highly pure allergens with a high batch-to-batch consistency satisfying the quality requirements of medicinal products manufactured by recombinant DNA technology can be produced. These reagents are expected to be qualitatively superior to the commercially available allergen preparations used for the in vitro and in vivo diagnosis of allergic conditions. In this article the current literature available on skin testing with such recombinant allergens (rAllergens) is reviewed and critically analyzed. To date many different rAllergens of various pollens, moulds, mites, bee venom, latex and celery have been used in skin testing in more than 1,600 allergic and control individuals. Skin prick tests as well as intradermal skin tests with rAllergens prove to be highly specific and safe. The diagnostic sensitivity of single rAllergens is generally lower than those obtained with allergen extracts, but can be considerably increased by using rAllergen panels covering the most important allergenic structures present in a given complex allergenic extract. Moreover, quantitative skin testing with single rAllergens allows interesting insights into correlations between the in vivo and in vitro sensitization to a given allergen. In conclusion, skin testing with rAllergens offers a highly specific and safe additional diagnostic tool to elucidate patient- and disease-specific sensitization patterns which will be needed for the development of patient-tailored immunotherapeutic treatments. PMID- 11435727 TI - Can immunoregulatory lactic acid bacteria be used as dietary supplements to limit allergies? AB - Studies in gnotobiotic animals have suggested that the intestinal bacterial flora may play an important role in priming the immune system during ontogeny to limit dysfunctional responses, including allergy. Prospective clinical studies have identified a higher incidence of allergy expression in early childhood among children who have low enteric populations of lactic acid bacteria (LAB), such as lactobacilli and bifidobacteria, further supporting a role for gut-colonizing bacteria in regulating immunological atopy. There is some evidence to suggest that supplementing the human diet with probiotic LAB might combat both allergy development and expression of atopy in allergy sufferers; however, definitive information, in the form of controlled intervention trials, remains scant. Recent immunological evidence has indicated that certain strains of LAB can stimulate the production of type I and II interferons and pro-interferon monokines (IL-12 and IL-18), following contact with the immune system; therefore, probiotic forms of immunoregulatory LAB could be used as dietary supplements to modify the gut microflora and provide pro-T helper cell 1 (Th1) STAT-activating signals sufficient to deviate the immune phenotype and correct the Th2-type bias which promotes allergy. This review outlines the clinical and laboratory evidence of a role for LAB in combating allergies, and attempts to explain this phenomenon in terms of our current understanding of immunoregulatory signals produced by gut colonizing microbes. PMID- 11435728 TI - Identification of a Hevea brasiliensis latex manganese superoxide dismutase (Hev b 10) as a cross-reactive allergen. AB - BACKGROUND: Cross-reactive allergens play an increasingly important role in latex allergy in complicating both the diagnosis and time course of allergic symptoms. Manganese superoxide dismutase (MnSOD), a ubiquitous protein of prokaryotic and eukaryotic organisms, was described as a cross-reactive allergen in Aspergillus fumigatus. Little information is available on the importance of this pan-allergen in Hevea brasiliensis latex. The aim of this study was to clone and express MnSOD from H. brasiliensis latex, and to obtain the soluble and immunologically active recombinant allergen for diagnosis of latex allergy and to investigate possible cross-reactivities with the structurally related A. fumigatus and human MnSODs. METHODS: A complementary DNA coding for Hevea latex MnSOD was amplified by PCR. The recombinant protein was produced in Escherichia coli with an N-terminal hexahistidyl tag. Enzymatic activity of the recombinant protein was determined using an enzyme assay for SODs. IgE immunoblotting and IgE inhibition assays were performed to characterize the recombinant allergen and its cross-reactivity. RESULTS: A Hevea latex MnSOD consisting of 206 amino acid residues was cloned and expressed in E. coli. The allergen was designated Hev b 10. The recombinant protein was enzymatically active, indicating the correct folding of the protein. In immunoblots, latex- as well as A. fumigatus-allergic patients revealed IgE binding to recombinant (r)Hev b 10. Cross-reactivity to Asp f 6, the MnSOD from A. fumigatus, and human MnSOD was determined by inhibition of IgE binding to these MnSODs by rHev b 10. CONCLUSIONS: Hev b 10 is a new cross-reactive allergen of H. brasiliensis which belongs to the 'latex-mold' group of latex allergens. Furthermore, it is a candidate for primary sensitization in patients allergic to the pan-allergen MnSOD. PMID- 11435729 TI - Characterization of high-molecular-mass allergens in oilseed rape pollen. AB - BACKGROUND: Oilseed rape pollen allergies have been previously described as the result of cross-sensitization with various pollens. Recently, several proteins have been identified as oilseed rape allergens. The aim of the present work was the characterization of oilseed rape pollen allergens by two-dimensional (2-D) gel analysis and amino acid microsequencing. METHODS: Water extractable proteins from oilseed rape pollen were separated by isoelectrofocusing and then transferred onto a nitrocellulose sheet. Twenty-one human sera from pollen- or mustard-allergic individuals were screened for their reactivity to oilseed rape proteins. Eleven sera possessed IgE which recognized oilseed rape pollen proteins and one serum was selected for further 2-D characterization and amino acid microsequencing of the allergens. RESULTS: The results showed that three molecules from oilseed rape pollen were identified as oilseed rape allergens which have not yet been described. These three proteins were molecules of 70 kD with a pI >8, 40 kD with a pI around 10 and 80 kD with a pI around 5. These proteins displayed identities with the berberine bridge protein, a receptor-like protein kinase and the cobalamin-independent methionine synthetase from Arabidopsis thaliana, respectively. The genes encoding the putative Arabidopsis molecules are located on chromosome 1 (berberine bridge protein) and chromosomes 3 and 4 (receptor-like protein kinases). CONCLUSION: These results show that certain high-molecular-mass proteins from oilseed rape pollen are allergens. PMID- 11435730 TI - Purification of the major allergen of red soft coral (Dendronephthya nipponica). AB - Red soft coral (RSC; Dendronephthya nipponica, a marine coelenterate) causes spiny lobster fishermen living along the Pacific coast of Miyazaki Prefecture in Japan to develop occupational allergies, such as conjunctivitis, rhinitis, dermatitis and bronchial asthma. The aim of this study was to purify and to characterize RSC allergen, which causes occupational asthma in spiny lobster fishermen. The allergic responsiveness of spiny lobster fishermen to RSC was examined. The examinations included specific IgE production, skin test responses, lymphocyte stimulation tests and specific IgG production. We found that RSC has a strong sensitizing activity in humans at a molecular weight of 10 kD or more, while it has no IgE-producing activity at a molecular weight of less than 10 kD. Neither the nonatopic controls nor the atopic non-coral-allergic controls exhibited any RAST-binding activity to any fraction. For the purification and the identification of this new allergen component, repeated gel filtration of the RSC extract was performed on a Sephacryl S-200 column, followed by gel filtration on a Superose-6 column. The purified major allergen component Den n 1, which is separated on a Mono-Q column, showed intradermal responses, lymphocyte stimulating activity and specific IgG-producing activity in RSC-induced bronchial asthma patients. The 53-kD component was electroblotted on a polyvinylidene difluoride membrane. The N-terminal amino acid sequence of this new allergen component (Den n 1) was determined as Asp-Asp-Ile-Asn-Arg-Tyr-Ala-Phe-Asp-Asn-Lys Ile-Asn- Asp-Lys-Leu-Phe-Asp-His-Trp-Gln-Ser. PMID- 11435731 TI - Recombinant Dirofilaria immitis-derived antigen can suppress passive cutaneous anaphylaxis reactions. AB - BACKGROUND: High levels of antigen-nonspecific IgE are produced in animals infected with helminth parasites. Generally, the increase in IgE is thought to exacerbate allergic reactions. However, high levels of antigen-nonspecific IgE may alter some features of anaphylactic reactions. To investigate the molecular mechanisms of antigen-nonspecific IgE production induced during filarial infections, we previously constructed rDiAg (recombinant Dirofilaria immitis derived antigen) in Escherichia coli. In the present study, we examined the effect of rDiAg on the production of antigen-nonspecific IgE and on allergic cutaneous reactions in rats. METHODS: Osmotic pumps filled with 200 microg of rDiAg or with 200 microl of PBS (control) were subcutaneously implanted in Wistar rats, and plasma samples were collected weekly thereafter. IgE levels were determined by ELISA. Homologous passive cutaneous anaphylaxis (PCA) reactions with anti-DNP-As IgE were examined 21 days after implantation. (125)I-IgE binding assays were examined on peritoneal mast cells from rDiAg-infused rats and control rats. RESULTS: Antigen-nonspecific IgE production was induced in rDiAg-infused rats. PCA reactions were suppressed in rDiAg-infused rats in spite of high levels of IgE and a markedly increased expression of Fc epsilon RI. (125)I-IgE did not bind to mast cells derived from rDiAg-infused rats, but it bound dose dependently to mast cells derived from control rats. CONCLUSION: The present data support the hypothesis that antigen-nonspecific IgE might protect against antigen-specific IgE by means of competition for mast cell receptors. rDiAg is an essential factor to induce antigen-nonspecific IgE in helminth infections. PMID- 11435732 TI - Characterization of alpha 5-integrin-dependent mast cell adhesion following Fc epsilon RI aggregation. AB - BACKGROUND: Integrin receptors are engaged in the upregulation of mast cell adhesion to extracellular matrix components upon stimulation with cytokines and antigen. Fibronectin receptor containing the alpha 5-integrin subunit is critical for mast cell interaction with the extracellular matrix protein fibronectin (FN). METHODS: The murine MCP5/L mast cell line was employed to investigate the process of Fc epsilon RI-mediated mast cell adhesion to FN. RT-PCR and cytofluorimetric analysis were used to assess the expression of alpha 5 integrin in MCP5/L mast cells. Radiolabelled mast cells were sensitized with monoclonal IgE and used in adhesion assays. Anti-alpha 5-integrin antibody (Ab), monovalent hapten and metabolic inhibitors were used to characterize antigen-mediated mast cell adhesion to FN. RESULTS: Addition of antigen to IgE-sensitized cells resulted in transient upregulation of mast cell adhesion to FN with a maximum adhesion following 30 min of incubation. Mast cell adhesion was inhibited with anti-alpha 5-integrin monoclonal antibodies blocking FN receptor or with excess monovalent hapten preventing antigen-mediated IgE cross-linking. The presence of the protein kinase C (PKC) inhibitor staurosporine also inhibited mast cell adhesion in a dose-dependent fashion. The process of Fc epsilon RI-mediated upregulation of mast cell adhesion to FN was not associated with an increase in surface expression of mast cell FN receptors. CONCLUSION: The major FN receptor on MCP5/L mast cell surface, an integrin containing the alpha 5 subunit mediates a transient change in mast cell adhesiveness following IgE cross-linking. Fc epsilon RI-derived signals engage PKC and upregulate mast cell adhesion in a process which might involve changes in integrin avidity rather than integrin expression. PMID- 11435733 TI - Preliminary evidence for 'aberrant' expression of the leukocyte integrin LFA-1 (CD11a/CD18) on conjunctival epithelial cells of patients with mite allergy. AB - BACKGROUND: We have previously shown aberrant expression of the 'leukocyte' integrin LFA-1 on epithelial cells in chronic autoimmune thyroiditis. In the present study we investigated whether conjunctival epithelial cells, which bear the adhesion molecule ICAM-1 on their surface during allergic inflammation, may also aberrantly express its natural ligand, the 'leukocyte' integrin LFA-1. METHODS: We studied 13 patients with rhinoconjunctivitis allergic to mites, chronically exposed to the allergen, 11 patients allergic to pollen tested out of the pollen season and 8 normal volunteers. Single and double immunocytochemical staining of conjunctival smears was employed. RESULTS: LFA-1 staining on epithelial cells was demonstrated in 12/13 patients allergic to mites and not in normal controls or in patients allergic to pollen tested out of the pollen season. The epithelial localization of LFA-1 was confirmed by double staining with anti-LFA-1 and anti-cytokeratin antibodies (both immunocytochemical and immunofluorescence). CONCLUSIONS: Coexpression of LFA-1 and ICAM-1 during persistent allergen stimulation may be relevant for interaction between epithelial cells and activated effector cells, such as eosinophils, which bear on their surface both ICAM-1 and its beta2 integrin ligands. PMID- 11435734 TI - Measurement of inflammatory mediators of mast cells and eosinophils in native nasal lavage fluid in nasal polyposis. AB - BACKGROUND: Nasal polyposis (NP) often coexists with asthma, rhinitis and sinusitis. Polyp histology typically shows chronic, eosinophilic inflammation. The inflammatory cell infiltrate generally includes eosinophils, lymphocytes, plasma cells and mast cells. OBJECTIVE: To gain insight into the natural history of NP, we analysed mediator levels and leukocyte values in nasal fluids and eosinophil cationic protein (ECP), total IgE levels and eosinophils in the blood in several groups of both allergic and non-allergic patients with nasal polyps and in patients with allergic rhinitis (AR). METHODS: Thirty-two patients with nasal polyps entered the study. As a control group, we studied 55 patients with AR, i.e. 20 patients with seasonal AR to grass pollen, 24 with AR sensitive to Parietaria and 11 with AR sensitive to house dust mite (HDM). Eighteen patients with nasal polyps were also allergic patients (8 were sensitive to Parietaria and 10 were sensitive to HDM), whereas 14 were non-allergic patients. Tryptase and histamine values were assayed in nasal fluids, whereas total IgE was determined in serum. ECP values were assayed both in nasal fluids and serum. Eosinophils were quantified both in the blood and nasal fluids. RESULTS: Tryptase levels were significantly higher in the nasal lavages from patients with NP than in those from patients without NP (4.7 vs. 3.5 U/l, p < 0.001) and correlated with symptom scores (r(s) = 0.42, p < 0.0001). The median levels of histamine in nasal fluids from patients with NP were also significantly higher than those observed in patients without NP (50.0 vs. 21.3 ng/ml, p < 0.001), but did not correlate with symptom scores. Finally, the median levels of ECP in nasal fluids from patients with NP were significantly higher than those observed in patients without NP (38.1 vs. 16.1 ng/ml, p < 0.001) and correlated with symptom scores (r(s) = 0.38, p < 0.001). Analysis of variance showed that, among the variables studied, the presence of nasal polyps was the factor responsible for the higher levels of tryptase, histamine and ECP in nasal fluids. With regard to leukocyte counts in nasal fluids, no significant differences were observed between rhinitis patients with NP and those without NP. With regard to serum ECP and serum total IgE, no significant differences were detected between the two groups under study. Blood eosinophil levels in patients with NP were significantly higher than those observed in patients without NP (5.8 vs. 5.6, p = 0.002). Analysis of variance showed that both the presence of nasal polyps and the type of sensitisation were important. Considering the total symptom scores, no significant differences were observed between rhinitis patients with NP and those without NP. CONCLUSIONS: The present findings are consistent with the view that chronic eosinophil mucosal inflammatory disease in NP involves a self-sustaining mechanism, i.e. local release of inflammatory mediators, independent of allergen stimulation of nasal mucosa. Increased release of inflammatory mediators contributes to the development of nasal polyps, determining oedema and an increased recruitment of inflammatory cells. Besides eosinophils, mast cells also play a key role in this process. PMID- 11435735 TI - Total serum IgE levels in chronic hepatitis C: influence of interferon alpha therapy. AB - BACKGROUND: Liver disease has been considered a prominent cause of IgE elevation. No data on serum IgE levels in chronic hepatitis C have been reported. Interferon alpha is a standard therapy for chronic hepatitis C. Cytokine use is a promising type of immunomodulation in the treatment of IgE-mediated diseases. The effects of interferon-alpha therapy on serum IgE have not been fully evaluated. The aim of the study was to evaluate both serum IgE levels in patients with chronic hepatitis C and the course of these levels after interferon-alpha therapy. PATIENTS AND METHODS: Serum IgE was determined in 100 adult patients with chronic hepatitis C (24 atopics according to positive skin prick tests and 76 nonatopics) and in 75 healthy controls (25 atopics and 50 nonatopics). Serum IgE measurements were repeated at 1 and 3 months of therapy with recombinant interferon-alpha (3 x 106 units s.c. 3 times weekly) in 34 of these patients. RESULTS: Serum IgE levels were similar in chronic hepatitis C patients and in controls when adjusted for atopic status. Among patients with chronic hepatitis C, serum IgE levels were unrelated to liver necroinflammatory activity. A modest but statistically significant increase of IgE values was observed after interferon-alpha therapy, particularly in patients with no virological response. CONCLUSIONS: Chronic hepatitis C is not a significant cause of increased total serum IgE values. Serum IgE increase in some patients with liver disease may be related to the cause of liver injury and not to liver disease per se. Interferon-alpha therapy in patients with chronic hepatitis C is followed by no modification or even a moderate increase of serum IgE values. PMID- 11435736 TI - Urticaria and rhinitis to shrubs of Ficus benjamina and breadfruit in a banana allergic road worker: evidence for a cross-sensitization between Moracea, banana and latex. AB - BACKGROUND: We report the case of a road worker with a food allergy to banana, who developed urticaria and rhinitis when cutting shrubs of Ficus benjamina and breadfruit. He did not develop an allergy to latex of Hevea brasiliensis. RESULTS: Sensitization to latex of F. benjamina, H. brasiliensis, breadfruit and banana was demonstrated using skin tests and specific IgE measurements. RAST inhibitions procedures showed that specific IgE to breadfruit latex cross-reacted more strongly with latex of H. brasiliensis and banana than with latex of F. benjamina with the same extract. CONCLUSION: Given the wide distribution of Moracea trees in tropical regions, sensitization to latex of H. brasiliensis and banana could be a consequence of sensitization to Moracea members; F. benjamina does not seem to be the only Moracea responsible for cross-allergy with latex and fruit. Consequently, it seems interesting to test other members of the Moracea family in patients sensitized to latex of H. brasiliensis and banana. Sensitization to breadfruit could be a risk factor for sensitization to latex of H. brasiliensis. PMID- 11435737 TI - Differential regulation of cathepsin B and prorenin gene expression in renal juxtaglomerular cells. AB - Cathepsin B (cathB) is a lysosome cysteine proteinase. It has been suggested to process prorenin to renin in the renin producing juxtaglomerular (JG) cells of the afferent arterioles (AA) of the kidney. Whether cathB expression is regulated similarly to prorenin production is yet unknown. We have measured prorenin and cathB mRNA levels as well as cathB protein levels in renal AA of Sprague-Dawley rats subjected to low (0.02% w/w) salt diet and ramipril treatment (10 mg/kg/day), or to normal (0.6%) or high (4%) salt diet. Prorenin and cathB expression were also analyzed in the JG cell line As4.1. Prorenin mRNA levels in animals on normal (plus ramipril), low or high salt diet correlated as 1:10:0.5, respectively, while cathB mRNA levels correlated as 1:1:0.6, respectively. Treatment of the As4.1 cells with 100 nM phorbol-12-myristate-13-acetate (PMA) for 16 h inhibited prorenin mRNA expression 3-fold relative to the control conditions. CathB mRNA abundance was not different between the PMA treated and the control As4.1 cells. Western analysis of the cathB protein abundance has shown no difference between the rats on normal and low salt diet, and decrease by 50% in the rats on high salt diet. The results of this study suggest that prorenin and cathB gene expression in renal JG cells are differentially regulated. PMID- 11435738 TI - Effects of angiotensin II and the AT(1) receptor antagonist losartan on the renal excretion of urodilatin. AB - BACKGROUND: The precise mechanisms regulating the natriuretic peptide urodilatin (ANP-95-126) remain to be defined. Renal excretion of urodilatin (U(URO)V) has been shown to be modified by variations in plasma sodium and renal perfusion pressure. This suggests a relationship between urodilatin and the renin angiotensin system. METHODS: We investigated the effects of angiotensin II (AII, 0.1 nmol/l) and the AT(1) receptor antagonist losartan (LS, 1 micromol/l) on U(URO)V and renal function in isolated rat kidneys perfused for 180 min in a closed circuit system. A further series employing a vasoconstricting concentration of endothelin-1 (ET-1, 0.01 nmol/l) was performed to explore the effects of vasoconstriction and glomerular filtration rate (GFR) on U(URO)V. RESULTS: Urine flow (UV) and urinary sodium excretion (U(Na)V) decreased and renal vascular resistance (RVR) increased after treatment with AII (n = 5) in comparison with a control group (n = 6; p < 0.05). Treatment with LS (n = 5) and AII+LS (n = 5) had no significant effect on these parameters. GFR decreased after AII (p < 0.05) and was not significantly altered by other interventions. U(URO)V decreased after AII (p < 0.05) and was comparable to the control group after LS and AII+LS. ET-1 (n = 5) induced a significant increase in RVR and decreased UV and U(Na)V (p < 0.05). Point-to-point analysis revealed that the ET-1-induced vasoconstriction and the subsequent decrease in GFR had no effect on U(URO)V. CONCLUSIONS: This suggests that vasoconstrictory concentrations of AII decrease U(URO)V in the isolated perfused rat kidney. The lack of effect of ET-1 on U(URO)V suggests that the AII-induced alterations in urodilatin excretion cannot be explained by vasoconstriction per se. PMID- 11435739 TI - High glucose levels alter angiotensin II-induced Ca(2+) uptake via PKC and cAMP pathways in renal proximal tubular cells. AB - Although a dysfunction of the calcium metabolism occurs in diabetes mellitus, alterations of Ca(2+) uptake induced by angiotensin II (ANG II) in renal proximal tubular cells (PTCs) grown in high-glucose medium are not fully elucidated. Thus, we examined whether high glucose concentrations can induce an alteration of the ANG II effect on the Ca(2+) uptake and its action mechanism in primary cultured renal PTCs. PTCs were exposed to different glucose concentrations (5-100 mM) and time intervals (0-48 h). There was a sustained increase of Ca(2+) uptake at glucose concentrations >15 mM. Thus, we selected 25 mM glucose and incubation for 48 h to maintain a hyperglycemic condition in vitro, unlike short-time regulatin. ANG II significantly inhibited the Ca(2+) uptake in a dose-dependent manner in a 5-mM glucose medium. In addition, downregulation of ANG II receptors appeared in a glucose dose dependent manner. However, PTCs treated with 25 mM glucose for 48 h, not 12 h, did not exhibit the inhibitory effect of ANG II (10(-7) M) on Ca(2+) uptake, although the inhibitory effect of ANG II on Ca(2+) uptake occurred in the presence of 25 mM mannitol or L-glucose. Staurosporine, bisindolylmaleimide I (protein kinase C, PKC, inhibitors), 12-o-tetradecanoylphorbol 13-acetate pretreatment, SQ 22536 (an adenylate cyclase inhibitor), and myristoylated protein kinase A inhibitor amide 14-22 (a protein kinase A inhibitor) blocked the 25-mM-glucose-induced alteration of ANG II effect on Ca(2+) uptake. These results suggest that both PKC and cyclic adenosine monophosphate (cAMP) pathways are involved in the high-glucose-induced alteration of ANG II effect on Ca(2+) uptake. Indeed, 25 mM glucose increased PKC activity and cAMP contents. In conclusion, a high glucose concentration altered ANG II induced inhibition of Ca(2+) uptake via PKC and cAMP pathways in the PTCs. PMID- 11435740 TI - Suppression of ICAM-1 expression in renal proximal tubular cells by 1,25 dihydroxyvitamin D3. AB - BACKGROUND: 1,25-Dihydroxyvitamin D(3) is mainly synthesized by renal proximal tubular cells. More recently, it has been shown to affect cell growth and TGF beta(1) synthesis in glomerular and tubular renal cells in vitro, and to prevent glomerulosclerosis in vivo in subtotally nephrectomized rats. The mechanisms involved have not been fully identified. We asked whether 1,25-vitamin D(3) might interact with additional immunoregulatory functions of renal cells by studying its effects on the expression of the cellular adhesion molecules ICAM-1 (CD54) and VCAM-1 (CD106) in human proximal tubular cells in vitro (HK-2 cells). METHODS: Expression of adhesion molecules was assessed in HK-2 cells cultured under basal conditions and after stimulation with TNF-alpha plus IFN-gamma, by flow cytometry, gene transcription (RT-PCR) and measurement of soluble ICAM-1 in culture supernatant by ELISA. RESULTS: Unstimulated HK-2 cells did not express VCAM-1 and only little ICAM-1. 1,25-Vitamin D(3) had no effect on the expression of adhesion molecules in unstimulated cells. TNF/IFN stimulation resulted in a 4 fold increase in ICAM-1 and VCAM-1 expression. The TNF/IFN-induced increase in ICAM-1 expression was reduced by 1,25-vitamin D(3) dose dependently (10(-7) M vs. solvent: -30%; 10(-9) M: -18%; 10(-11) M: -17%). 25(OH)-vitamin D(3) had no effect. ICAM-1 mRNA concentration was increased in TNF/IFN-stimulated cells. 1,25 Vitamin D(3) treatment prevented the increase of ICAM-1 mRNA by 27% after 24-72 h incubation (p = 0.03). The TNF/IFN-induced increase in soluble ICAM in culture supernatants was unchanged by 1,25-vitamin D(3). VCAM-1 expression was unchanged by incubation with 1,25-vitamin D(3) under basal conditions and after TNF/IFN stimulation. CONCLUSION: 1,25-Vitamin D(3) inhibits cytokine-induced ICAM-1 but not VCAM-1 expression in renal proximal tubular cells in vitro. The present data support the hypothesis that 1,25-vitamin D(3) is not only synthesized by renal tubular cells, but may also affect immunoregulatory functions in these cells. PMID- 11435741 TI - Podocyte injury predicts prognosis in patients with iga nephropathy using a small amount of renal biopsy tissue. AB - To predict the progression in patients with IgA nephropathy, we analyzed glomerular lesions except for sclerosis, adhesion and/or crescents in 34 patients with this disease by morphometric analysis. Levels of urinary protein excretion (UP), creatinine clearance (Ccr), serum creatinine (sCr) and mean blood pressure (MBP) at the time of renal biopsy were used as the clinical parameters. The slope of 1/sCr was also used as a prognostic parameter. Renal specimens were obtained by echo-guided biopsy. In PAS-stained light microscopic renal sections, three mid sections of open glomeruli were selected and photographed. Stereologic estimation was performed as follows: absolute values of glomerular volume (V(G)), glomerular surface area (S(G)), podocyte and nonpodocyte cell number per glomerulus (N(G(pod)) and N(G(Non-pod))), glomerular surface area covered by one podocyte S(G)/N(G(pod))) and glomerular volume occupied by one nonpodocyte cell (V(G)/N(G(Non-pod))). There was a significant correlation between the levels of UP and the change of podocyte injury parameters (N(G(pod)) and S(G)/N(G(pod))) or N(G(Non-pod)). N(G(pod)) was negatively but S(G)/N(G(pod)) and N(G(Non-pod)) were positively correlated with UP. S(G)/N(G(pod)) or N(G(Non-pod)) was correlated with MBP. N(G(pod)), S(G)/N(G(pod)), N(G(Non-pod)), UP or MBP was significantly correlated with the slope of 1/sCr. High specificity was observed for N(G(pod)), S(G)/N(G(pod)) and MBP. High sensitivity was also observed for N(G(Non-pod)) and UP. It appears that podocyte injury might provide additional prognostic information in patients with IgA nephropathy. PMID- 11435742 TI - Absence of cytokine response to bacterial challenge in human tubuloepithelial cells. AB - In acute bacterial renal infections, which are most frequently caused by Escherichia coli, tubuloepithelial cells are involved with respect to bacterial adherence, invasion and cytotoxicity. In addition, cytokines expressed by tubuloepithelial cells may be relevant for the recruitment of inflammatory cells and tissue damage in bacterial interstitial nephritis. We asked which inflammatory cytokines are produced by primary human tubuloepithelial cells following in vitro exposure to E. coli and found no release of IL-6, IL-8 and TNF alpha by tubular cells challenged by bacteria. Purified virulence factors (fimbriae, lipopolysaccharide) from E. coli were also without effects on cytokine release by tubular cells. Since lymphocytic infiltration is a characteristic feature in the chronic form of interstitial nephritis, MHC class II expression by tubular cells in response to bacterial coincubation was analyzed. Exposure to both IFN-gamma and E. coli enhanced MHC class II expression on tubuloepithelial cells. In conclusion, tubuloepithelial cells may play an active role in the local defense against bacteria, e.g. by expressing MHC class II molecules. However, in vitro inflammatory cytokines are not induced by E. coli in this cell population. PMID- 11435743 TI - Pathophysiology of cyclosporine-induced nephrotoxicity in humans: a role for nitric oxide? AB - BACKGROUND: The causes for the nephrotoxicity of cyclosporine A (CsA) have not been fully elucidated. Intrarenal vasoconstriction induced by several different mediators, both in humans and experimental animals, have been proposed. METHODS: We studied prostaglandin metabolites, endothelin and nitric oxide in kidney transplant patients receiving their first CsA dose. Prostaglandin metabolites in the urine and endothelin and nitric oxide (NO2/NO3 in urine and plasma were measured in 14 patients before and 3 and 6 h after oral ingestion of CsA (10 mg/kg b.w.). Clearances for inulin and p-aminohippuric acid (PAH) were measured before and in two separate 3-hour periods after CsA. Blood pressure, heart rate, and CsA blood levels were also determined. RESULTS: Clearances of inulin and PAH decreased progressively after CsA dosage while renal vascular resistance increased. Nitric oxide plasma levels decreased in nearly all patients from 21.0 +/- 2.8 to 19.1 +/- 2.6 (p = 0.003) and then rose slightly to 19.5 +/- 2.5 micromol/l (p = 0.1) 3 and 6 h after CsA ingestion, respectively. Urinary excretion of NO2/NO3 decreased nonsignificantly from 269 +/- 38.8 to 259 +/- 27.7 and 254 +/- 41.6 micromol/min (p = 0.5 and 0.5). At the same time, urinary prostaglandin E2 and 6-keto-prostaglandin F(1 alpha) excretion rate declined significantly [from 1,187 +/- 254 to 1,186 +/- 351 and 730 +/- 148 pg/min (p = 0.27 and 0.02) and from 697 +/- 115 to 645 +/- 134 and 508 +/- 58.2 pg/min (p = 0.34 and 0.05)]. Urinary thromboxane B2 and plasma and urinary endothelin first increased and then decreased nonsignificantly. Mean arterial pressure rose from 107 +/- 2.5 to 110 +/- 2.6 and 114 +/- 3.4 mm Hg (p = 0.1 and 0.05). CONCLUSION: The pathophysiology of CsA-induced acute renal vasoconstriction involves several different mechanisms including a decrease of the vasodilating prostaglandins E2 and 6-keto-prostaglandin F(1 alpha) and possibly nitric oxide. PMID- 11435744 TI - Oxidative stress and renal dysfunction in salt-sensitive hypertension. AB - Hypertension is a risk factor for the development of end-stage renal disease. The mechanisms underlying hypertensive nephropathy are poorly understood. There is evidence, however, that in hypertension there is an accumulation of partially reduced oxygen and its derivatives, known collectively as reactive oxygen species, which may contribute to progressive renal dysfunction. In the present study, we assess the contribution of oxidative stress in the development of salt dependent hypertensive nephrosclerosis. Going beyond previous end point studies, which inferred renal function either indirectly or only qualitatively, we have determined oxidative stress concurrently with direct and quantitative measurements of renal function (via inulin and p-aminohippuric acid clearances). Moreover, in this time-dependent study, the measurements have been taken under low- as well as high-salt diets. As was expected from previous studies, in the Dahl salt-sensitive rat, a high-salt diet (8% NaCl) resulted in the development of hypertension, in a decreased glomerular filtration rate, and in a decreased renal plasma flow as compared with the normotensive control, the Dahl salt resistant rat. In addition, however, we found clear evidence for the accumulation of reactive oxygen species in renal tissue homogenates of Dahl salt-sensitive rats on the high-salt diet. Our time-dependent protocol also indicated that renal oxidative stress follows, in time, the development of hypertension. We also found that after 2 weeks of increased salt loading, Dahl salt-sensitive rats excreted less cyclic guanosine monophosphate and NO(x) than Dahl salt-resistant rats on the same diet. It is known that urinary cyclic guanosine monophosphate and NO(x) represent the activity and stable derivatives of renal NO., respectively, and that they closely correlate with renal vascular resistance. Therefore, our results suggest that, in the Dahl salt-sensitive rat, increased oxidative stress is associated with salt-dependent hypertensive nephrosclerosis and that decreased NO. bioavailability may represent a common factor responsible for the vascular and glomerular dysfunction. PMID- 11435745 TI - Factor analysis, including antihypertensive medication, of the outcome of pregnancy in pregnancy-associated hypertension. AB - AIM: To study the influence of different maternal factors, including antihypertensive medication, on the outcome of pregnancy in primi- and multiparas with pregnancy-associated hypertension. METHODS: A retrospective, multiple variate analysis was undertaken of the influence of several maternal factors, including antihypertensive medication, on fetal death and Apgar scores and the correlation between the medication and the number of caesarean sections in 127 episodes of pregnancy-associated hypertension was studied for the whole group as well as for primi- and multiparas separately. Of the multiparas, 40.8% had a history of preeclampsia, 19.7% of chronic hypertension and 9.2% of diabetes mellitus. Antihypertensive treatment aimed at achieving a blood pressure of 140/90 mm Hg. Forty-one patients (32.3%) received intravenous hydralazine, 25 (19.7%) received nifedipine per os and 44 (34.6%) received labetalol per os. RESULTS: The maximum systolic and diastolic blood pressure in the patients given intravenous hydralazine, nifedipine per os or labetalol per os did not differ, whereas in the multiparas, the number of patients who reached the target blood pressure while using labetalol was higher than with the two other medications, especially in comparison with intravenous hydralazine. For the primiparas, the time of delivery was the only factor with a significant impact on the Apgar scores. In the multiparas, there was an additional negative influence of the use of intravenous hydralazine. This was not seen in the patients using nifedipine and labetalol. CONCLUSIONS: The results suggest that in multiparas, intravenous hydralazine is possibly associated with more fetal distress when compared to primiparas. PMID- 11435746 TI - Lesions in lateral hypothalamic areas increase splenocyte apoptosis. AB - OBJECTIVE: The lateral hypothalamic area (LHa) is involved in various functions such as feeding, drinking, sexual and reward behavior, among others. Recently, we demonstrated that the LHa can regulate cellular immunity in the spleen. In experiments involving the LHa, it was noticed that the spleen shrinks noticeably after LHa destruction. To explore this phenomenon further, the effect of LHa lesioning on splenocyte apoptosis was investigated. METHODS: Male Wistar-King Aptekman rats underwent bilateral lesioning of their LHa and consequent spleen weights, splenocyte numbers and apoptosis were measured. For the detection of splenocyte apoptosis, both ELISA, which measures DNA fragmentation within the splenocytes, and flow cytometry, which measures the percentage of apoptotic lymphocytes in the spleen, were used. RESULTS: In the LHa-lesioned rats, spleen weights and the number of splenocytes decreased significantly within 24 h. Additionally, in the spleen, lymphocyte apoptosis significantly increased compared to the control after 6 h. CONCLUSION: These results suggest that the LHa may play a role in immunoregulation by affecting lymphocytes in the spleen through apoptosis and may be relevant to the pathway of stress-induced apoptosis. PMID- 11435747 TI - Immunomodulatory effects of cold stress on mice infected intraperitoneally with a 50% lethal dose of Toxoplasma gondii. AB - Cofactors such as stress have been suspected to play a role in the susceptibility to opportunistic infections. Toxoplasma gondii is one of the major opportunistic infectious agents in immunocompromised individuals, and infection can be modulated by external factors such as stress. OBJECTIVE: The purpose of this study was to examine the in vivo and in vitro role of cold stress (CS) in the pathogenesis of T. gondii infection and its impact on regulatory cytokines in this model. METHODS: Mice subjected to CS and control animals were infected intraperitoneally with an LD(50) of PD2 T. gondii tachyzoites, and the outcome of the infection was determined. In addition, peritoneal macrophages obtained from CS and non-stressed mice were infected in vitro with T. gondii. The number of infected macrophages, the number of intracellular parasites and the production of interferon (IFN)-gamma, interleukin (IL)-12, and tumor necrosis factor (TNF) alpha were determined. RESULTS: CS applied before intraperitoneal inoculation increased susceptibility against T. gondii infection. Peritoneal cells from CS mice contained significantly higher numbers of intracellular parasites and infected macrophages compared to those from non-stressed animals. IFN-gamma production was initially high in the CS group but decreased significantly after 36 h. Opposite results were found in the non-stressed group. Macrophages from CS mice persistently produced high levels of TNF-alpha and IL-12 and peaked after 36 h. Levels of these cytokines were lower or absent in the non-stressed group. CONCLUSION: These results suggest that CS increased the host susceptibility to intraperitoneal T. gondii infection by modulating the function of macrophages and the production of cytokines (IFN-gamma) involved in the early control of infection. PMID- 11435748 TI - Role of capsaicin-sensitive afferents in fever and cytokine responses during systemic and local inflammation in rats. AB - OBJECTIVE: Peripheral afferents play an important role in fever. In the present study, we investigated the role of capsaicin-sensitive afferents in fever and cytokine responses during systemic (induced by intraperitoneal lipopolysaccharide, LPS) and local (induced by injection of Freund's incomplete adjuvant, FIA, into the paw) inflammation. METHODS: Fevers in rats (8-10 weeks of age) whose capsaicin-sensitive afferents were depleted by neonatal capsaicin (50 mg/kg) treatment were compared to those of rats treated as neonates with vehicle. To investigate a possible involvement of cytokines, plasma levels of interleukin 6 (IL-6) and tumor necrosis factor (TNF) were measured during LPS- and FIA induced fever in rats after capsaicin-induced desensitization. Body temperature was measured by biotelemetry. IL-6 and TNF bioactivities in plasma were determined using bioassays. RESULTS: The initial but not the late phase of LPS (50 microg/kg)-induced fever was markedly higher (approximately 1.0 degree C) in rats whose capsaicin-sensitive neurons were destroyed by neonatal capsaicin treatment. Capsaicin-induced desensitization also resulted in significantly higher plasma levels of IL-6 and TNF 1 but not 4 h after LPS challenge. In contrast, the day after injection with FIA (0.1 ml), rats treated with capsaicin had significantly lower body temperatures compared with vehicle-treated animals. No differences were found in plasma levels of IL-6 and TNF between capsaicin- and vehicle-treated animals in response to FIA. CONCLUSIONS: These data indicate that the role of capsaicin-sensitive afferents in fever depends on the type of inflammatory response. During systemic inflammation, capsaicin-sensitive afferents may be involved in modulating fever by regulating the levels of pyrogenic cytokines. During local inflammation, the late phase of fever is partially mediated via capsaicin-sensitive afferents. PMID- 11435749 TI - Physiological concentrations of dopamine inhibit the proliferation and cytotoxicity of human CD4+ and CD8+ T cells in vitro: a receptor-mediated mechanism. AB - OBJECTIVE: Dopamine, a catecholamine neurotransmitter, influences growth and proliferation of lymphocytes. Pharmacological doses of dopamine have been shown to modulate T cell functions significantly, but no information is available on the effect of physiological concentrations of circulating dopamine on CD4+ and CD8+ T cell functions. This information may be of importance since significantly elevated plasma dopamine levels were observed in humans during uncoping stress, and suppression of T cell functions during stress is a well-known phenomenon. However, the mechanism inducing the suppression of T cell functions during stress is not yet clear. In the present investigation, we evaluated the effect of the dopamine level attained in the plasma of individuals with uncoping stress on the proliferation and cytotoxicity of CD4+ and CD8+ T cells in vitro. METHODS: T cell subpopulations were separated by panning. The effect of dopamine on IL-2-induced cell proliferation in vitro was evaluated by [3H]thymidine incorporation and cytotoxicity by 51Cr release, receptors by radioligand binding, cAMP by an assay kit and apoptosis by DNA fragmentation. RESULTS: At these elevated physiological concentrations, dopamine was found to inhibit significantly the proliferation and cytotoxicity of CD4+ and CD8+ T cells in vitro. This dopamine-mediated inhibition of proliferation was more marked on CD8+ T cells than on CD4+ T cells. The underlying mechanism was found to be D1 class of dopamine-receptor-mediated stimulation of intracellular cAMP. CONCLUSION: Results may be of significance to understand the role of peripheral dopamine in human neuroimmune communication in terms of physiological homeostasis in health and disease. PMID- 11435750 TI - Effects of rotational stress of different duration on NK cell activity, proinflammatory cytokines, and POMC-derived peptides in mice. AB - Previous studies have shown that the same stressor, depending on intensity, controllability, or duration, can have different effects on the immune system. The purpose of this study was to determine the effect of 10- and 20-min rotation on natural killer (NK) cell activity and also to establish if changes in body temperature, proinflammatory cytokine (IL-1beta, IL-6, and TNF-alpha) levels, and proopiomelanocortin (POMC)-derived peptide (ACTH and beta-endorphin) levels parallel the changes in NK cell activity in mice. We found that 10-min rotation significantly increased NK cell activity as compared to both the control (home cage) group and the 20-min-rotation group, while NK cell activity in the 20-min group was not significantly changed compared to the control group. Both 10 and 20 min of rotational stress decreased body temperature and induced significant changes in the proinflammatory cytokine and POMC-derived peptide levels as compared to the control group. The pattern of proinflammatory cytokine expression was quite different between the 10- and 20-min rotation groups. All three proinflammatory cytokines were expressed sequentially (at 0 h after rotation TNF alpha, at 6 h IL-1beta and IL-6, and at 24 h IL-6) in the 10-min rotation group, while the 20-min rotation group had a small increase in IL-1beta (6.7 +/- 1.8 pg/ml) at 0 h and increased levels of IL-6 at 6 and 24 h. There was a dissociation of ACTH and beta-endorphin expression in both groups resulting in significantly more beta-endorphin (p < 0.05) in the 10-min group at 6 h and significantly more ACTH (p < 0.04) in the 20-min group at 6 h. IL-1beta and beta endorphin have both been shown to have a direct stimulatory effect on NK cell activity. Therefore, we suspect that the significant increase in both IL-1beta and beta-endorphin at 6 h in the 10-min-rotation group may be involved in the increased NK cell activity observed at 24 h in the 10-min-rotation group. PMID- 11435751 TI - Maternal undernutrition induces neuroendocrine immune dysfunction in male pups at weaning. AB - The present study was designed to assess the effect of maternal undernutrition, during gestation and lactation, on the neuroendocrine [hypothalamo-pituitary adrenal (HPA)]-immune axis response to endotoxin (LPS) administration. For this purpose, 21-day-old male rats from both well-nourished (WN) and undernourished (UN) mothers were examined 2 h after injection (i.p.) of vehicle alone (VEH) or containing LPS (130 microg/kg BW). Circulating levels of glucose (GLU), ACTH, corticosterone (B), tumor necrosis factor-alpha (TNFalpha) and leptin were explored. The results indicate that: (a) mother body weight was significantly (p < 0.05) reduced, as a consequence of UN, at the second and third weeks of pregnancy; (b) no differences in basal glycemia were found in the two groups of pups, and LPS treatment did not induce hypoglycemia, in either group; (c) basal plasma ACTH, B and TNFalpha levels were similar in the two groups, and LPS induced ACTH, B and TNFalpha secretions, although severalfold higher than respective VEH values (p < 0.05) in pups from WN mothers, were fully (ACTH and B) and partially (TNFalpha) abolished in products from UN mothers; (d) both mean body weights and basal plasma leptin levels were significantly (p < 0.05) lower in pups from UN than from WN mothers, and LPS administration did not modify plasma leptin values in products from both groups. In addition, results of dispersed total adrenal cells incubated in vitro indicate that: (a) both basal and ACTH (22 pM)-induced B secretion were significantly (p < 0.05) lower in cells from UN than WN pups, and (b) leptin (100 nM) was able to inhibit partially ACTH stimulated B output by adrenal gland (AG) cells from WN pups; however, it failed to inhibit ACTH-stimulated glucocorticoid release by AG cells from UN pups. The present results indicate that undernutrition in mothers, during the very critical periods of gestation and lactation, induces in their male pups at weaning: (a) reduced circulating leptin levels and body weight values; (b) metabolic adaptation to normal carbohydrate metabolism; (c) hyporesponsiveness of the HPA and immune (TNFalpha) axes during endotoxemia, and (d) leptin resistance at the adrenocortical level. This study strongly supports that undernutrition of mothers results in neuroendocrine immune dysfunction of their pups; however, adrenal resistance to the inhibitory effect of leptin on glucocorticoid output is developed, probably as an adaptive mechanism to counteract unfavorable metabolic conditions. PMID- 11435752 TI - Concentration of soluble adhesion molecules (sVCAM-1, sICAM-1 and sL-selectin) in the cerebrospinal fluid and serum of patients with multiple sclerosis and systemic lupus erythematosus with central nervous involvement. AB - OBJECTIVES: The aim of the present study was to investigate the role of soluble adhesion molecules in the pathogenesis of multiple sclerosis (MS) and systemic lupus erythematosus (SLE) with demyelinating syndrome. METHODS: Paired cerebrospinal fluid (CSF) and serum samples were analysed by an ELISA method to determine the concentrations of sVCAM-1, sICAM-1 and sL-selectin. Intrathecal syntheses of the adhesion molecules were calculated. RESULTS: Elevated serum and CSF concentrations of sVCAM-1 were present in all patient groups. Intrathecal synthesis of sVCAM-1 was present in the relapsing-remitting and secondary progressive forms of MS. Intrathecal synthesis of sICAM-1 was observed in all clinical forms of MS. MS patients with progressive forms of the disease and SLE patients were characterised by intrathecal synthesis of sL-selectin. CONCLUSIONS: The data presented suggest that (1) blood-brain barrier damage can be assumed both in systemic disease and organ-specific disease (sVCAM-1), (2) clinical forms of MS differ from each other in respect to concentrations of adhesion molecules and (3) similar immunological events in the central nervous system of SLE patients with demyelinating syndrome and progressive forms of MS can be assumed (sL-selectin). PMID- 11435753 TI - Aging-related increased expression of inducible nitric oxide synthase and cytotoxicity markers in rat hypothalamic regions associated with male reproductive function. AB - We have previously demonstrated that the inducible nitric oxide synthase (iNOS) protein and total NOS activity increase in the hypothalamus and other regions of the male rat brain during aging. We have now tested the hypothesis that increased iNOS results in excessive nitric oxide (NO) and peroxynitrite production, and leads to increased apoptosis in CNS cells, including the GnRH and oxytocin hypothalamic neurons involved in the control of male reproductive function. Young (3-month-old) and old (24-month-old) male Brown Norway rats (n = 6) were perfused with 4% formalin. Adjacent coronal paraffin-embedded sections (5 microm) of preoptic area (POA), supraoptic nucleus (SON), paraventricular nucleus (PVN), and arcuate nucleus (ARC) of the hypothalamus were immunostained with antibodies for iNOS, neuronal NOS (nNOS), and nitrotyrosine (a marker of peroxynitrite formation). The intensity of immunostaining was measured using a densitometric image analysis system. Apoptosis was determined by the TUNEL assay. Double immunofluorescence staining with confocal laser scanning microscopy was used for co-localization studies. A significant increase in the iNOS immunostaining measured as optical density (OD) was found in the old compared to the young animals (SON: 0.32 +/- 0.02 vs. 0.23 +/- 0.03, p < 0.05; PVN: 0.34 +/- 0.03 vs. 0.07 +/- 0.05, p < 0.001; POA: 0.18 +/- 0.02 vs. 0.01 +/- 0.02, p < 0.001). Aging did not affect nNOS expression. Nitrotyrosine was elevated in the hypothalamic regions of old compared to young rats (SON: 0.32 +/- 0.05 vs. 0.10 +/- 0.04, p < 0.05; PVN: 0.32 +/- 0.04 vs. 0.13 +/- 0.03, p < 0.01; POA: 0.72 +/- 0.06 vs. 0.03 +/- 0.003, p < 0.001). Increased nitrotyrosine was accompanied by an elevation of the apoptotic index in the old rats (SON: 11.01 +/- 3.33 vs. 0.57 +/- 0.50, p < 0.001; PVN: 3.08 +/- 1.12 vs. 0.42 +/- 0.32; POA: 6.60 +/- 1.93 vs. 0.18 +/- 0.17, p < 0.01; ARC: 0.001 +/- 0.0001 vs. 4.33 +/- 2.33). iNOS staining co localized with GnRH and oxytocin staining. IN CONCLUSION: The aging-related iNOS increased expression in the hypothalamus of the male rat affects regions known to control the synthesis and release of GnRH (POA, ARC) and oxytocin (PVN, SON), and the factors regulating penile erection (POA, and PVN). These observations suggest that iNOS may play a role in the reduction in GnRH and oxytocin neuronal secretion resulting in reproductive dysfunctions such as lowered serum testosterone, hypospermatogenesis, and diminished copulatory function in the aging male animal. PMID- 11435754 TI - A reassessment of leptin's role in triggering the onset of puberty in the rat and mouse. AB - Leptin is an adipocyte-derived hormone that has been implicated to serve as a metabolic signal to the reproductive axis. The role of leptin in pubertal maturation, however, has been a much-debated topic. We have previously reported that leptin serves as a permissive signal to the onset of puberty in the female rat. In an attempt to further understand the mechanics of leptin during pubertal maturation in rodent species, we had three experimental objectives: first, to describe the temporal relationship of leptin with development in the male and female rat; second, to seek evidence for an increase in responsiveness of the neuroendocrine axis to leptin by assessing for possible changes in leptin receptor expression during pubertal developmental in the female rat; and, third, to reevaluate the possible role of leptin as a permissive signal to the onset of puberty in the mouse. We found that serum leptin levels remain relatively constant during the prepubertal and postpubertal stages of both sexes. In addition, we could not detect any significant developmental changes in leptin receptor gene expression in the hypothalamus of the female rat. Lastly, we corroborated our findings in the female rat that leptin reversed the delay in pubertal maturation secondary to food restriction but did not advance the onset of puberty in female mice. Together, these results suggest that leptin is not a metabolic trigger for the onset of puberty in the rodent; instead, leptin is one of several permissive factors, whose presence may be necessary but alone is not sufficient to initiate sexual maturation in these species. PMID- 11435755 TI - Estrogen and laryngeal synaptic strength in Xenopus laevis: opposite effects of acute and chronic exposure. AB - Synaptic transmission at the vocal synapse, the laryngeal neuromuscular junction, of Xenopus laevis has been shown to be regulated by long-term changes in circulating estrogen. In females, high levels of circulating estrogen also accompany gonadotropin-induced ovulation and oviposition and the switch from sexually unreceptive to receptive states, including changes in vocal behaviors (ticking to rapping). Here we examine the effects of gonadotropin injection on laryngeal synaptic strength and call type. Gonadotropin acutely reduced quantal content values of laryngeal synapses in intact, adult females; the lowest values were attained by 12 h post-injection. Estrogen and progesterone levels increased following human chorionic gonadotropin (hCG) injection; the time course was similar to, but negatively correlated with, changes in synaptic strength. In ovariectomized frogs, exogenous estrogen, but not progesterone or hCG, mimicked the acute effects of hCG in weakening laryngeal synapses of intact frogs. hCG injection suppressed ticking and sometimes induced rapping. Females could tick with either strong or weakened laryngeal synapses while rapping was only produced during the weakening action of hCG. The normally strong synapses of females may enable vocal production even when laryngeal synapses are weakened by hormones that induce ovulation. In contrast to the acute effect of estrogen on weakening laryngeal synapses, juveniles required more than 2 weeks of estrogen treatment to strengthen laryngeal synapses while at least 4 weeks postovariectomy were required to weaken synapses in adult females. We conclude that acute (hours) increases in circulating levels of estrogen weaken synapses while chronic (weeks) increases strengthen laryngeal synapses. PMID- 11435756 TI - Homologous upregulation of sst2 somatostatin receptor expression in the rat arcuate nucleus in vivo. AB - In vitro studies using various cell systems have provided conflicting results regarding homologous regulation of somatostatin (SRIH) receptors, and information on whether SRIH regulates the expression of its own receptors in vivo is lacking. In the present study we examined, by in situ hybridization, the effects of pretreatment with the sst2-preferring SRIH analog, octreotide, in vivo, on mRNA levels of two SRIH receptor subtypes, sst1 and sst2, in rat brain and pituitary. (125)I-[DTrp(8)]-SRIH binding was also measured in these regions. Three hours after the iv injection of 50 microg octreotide to conscious adult male rats, there was a 46% increase (p < 0.01) in the labeling density of sst2 mRNA expressing cells in the hypothalamic arcuate nucleus compared to normal saline pretreated controls, but not in any of the other brain regions examined. Computer assisted image analysis revealed that 3 h exposure to octreotide significantly (p < 0.01) augmented both the number and labeling density of sst2 mRNA-expressing cells in the arcuate nucleus, compared to those in saline-treated controls. By contrast, within the anterior pituitary gland, in vivo exposure to octreotide did not affect the expression of sst2 mRNA. No changes in sst1 mRNA-expressing cells were observed after octreotide treatment in any of the regions measured, indicating that the observed effects were homologous, i.e. specific of the receptor subtype stimulated. Octreotide pretreatment was also without effect on the density of (125)I-[DTrp(8)]-SRIH binding in either the arcuate nucleus or pituitary. These results demonstrate, for the first time, that SRIH preexposure in vivo upregulates the expression of a subtype of its own receptors, sst2, within the central nervous system. They further suggest that pretreatment with SRIH in vivo does not cause sst2 receptor desensitization in arcuate nucleus and pituitary. Such homologous regulatory mechanisms may play an important role in the neuroendocrine control of growth hormone (GH) secretion by the arcuate nucleus. PMID- 11435757 TI - Rapid stimulatory effects of brain-derived neurotrophic factor and neurotrophin-3 on somatostatin release and intracellular calcium rise in primary hypothalamic cell cultures. AB - Although the long-lasting effects of neurotrophins have been extensively studied, less data are available on their rapid effects, especially on peptide release. In the present report, we investigated rapid effects of neurotrophins on somatostatin release and on intracellular calcium concentration ([Ca(2+)](i)) in primary cultures of hypothalamic neurons. RT-PCR experiments revealed mRNA expression of the three high-affinity neurotrophin receptors tyrosine kinase (Trk) TrkA, TrkB and TrkC, indicating potential responses to their preferential ligands: nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT-3), respectively. We demonstrated that BDNF, and to a lesser extent NT-3, induced significant time- and concentration-dependent somatostatin release, while NGF was devoid of any effect. BDNF or NT-3 induction of somatostatin release was inhibited by the Trk inhibitors K-252a and genistein, whereas K-252b, a less effective inhibitor, had no effect. BDNF- and NT-3-induced somatostatin release depended upon extra- and intracellular Ca(2+) since it was completely abolished in the presence of the Ca(2+) chelators BAPTA (bis-(alpha aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid) or BAPTA-AM (bis-(alpha aminophenoxy)-ethane-N,N,N',N'-tetraacetoxymethylester), respectively. In addition, BDNF and NT-3 induced a sustained and rapid increase in [Ca(2+)](i) which depended on the extracellular Ca(2+) concentration. MK-801 (dizocilpine) and tetrodotoxin (TTX) entirely blocked neurotrophin-evoked somatostatin release and [Ca(2+)](i) rise in response to BDNF and NT-3 application in most neurons. Neurotrophin-induced [Ca(2+)](i) rise was completely blocked by K-252a. The present results are consistent with: (1) an indirect effect of neurotrophins on somatostatin release via endogenous glutamate release and subsequent NMDA receptor activation, (2) a major indirect effect of neurotrophins on Ca(2+) rise in hypothalamic neurons which very likely occurs through NMDA receptor activation. Taken altogether, these results indicate that BDNF and NT-3 can rapidly affect the activity of hypothalamic neurons. PMID- 11435758 TI - The circadian clock, light/dark cycle and melatonin are differentially involved in the expression of daily and photoperiodic variations in mt(1) melatonin receptors in the Siberian and Syrian hamsters. AB - Mechanisms underlying the daily and photoperiodic variations in mt(1) melatonin receptors were investigated in the pars tuberalis (PT) and suprachiasmatic nuclei (SCN) of Siberian and Syrian hamsters. Whatever its daily profile, melatonin receptor density was strongly increased in both structures and species after constant light exposure or pinealectomy, and decreased after a single melatonin injection, indicating melatonin involvement in the daily regulation of the receptor protein. This was confirmed by a strong inverse correlation between melatonin binding capacity and plasma melatonin concentration. In contrast, regulation of mt(1) mRNA appeared more complex. The circadian clock, the light/dark cycle and melatonin are all implicated in mt(1) gene daily fluctuations, but the extent of their involvement depends upon the structure and the species studied. The photoperiodic decrease in melatonin receptor density observed in short photoperiod (PT of the two hamster species and Syrian hamster SCN) seems to be the consequence of a long-term mt(1) gene repression induced by the lengthening of the melatonin peak. Altogether, these results show that during daily variations, mt(1) melatonin receptor mRNA and protein are differentially regulated, while at the photoperiodic level, the mt(1) protein status depends on mRNA transcription. PMID- 11435759 TI - Regulation of norepinephrine in the medial preoptic area of Siberian hamsters by gonadal steroids. AB - Photoperiod has profound effects upon the neuroendocrine axis underlying reproductive physiology in seasonally breeding mammals. For long-day (LD) breeders, such as the Siberian hamster, exposure to a short-day (SD) photoperiod results in declines in circulating levels of gonadal steroids, luteinizing hormone (LH), and prolactin (PRL). The current study sought to investigate the effects of photoperiod and steroid levels on norepinephrine (NE), one of the major neurochemical regulators of gonadotropin-releasing hormone (GnRH) function. Since NE release within the medial preoptic area (mPOA) has been shown to stimulate the activity of GnRH cells, it was hypothesized that exposure to a short photoperiod would decrease NE levels. Furthermore, since gonadal steroids show negative feedback on GnRH function, it was hypothesized that gonadectomy would result in increased levels of NE. Adult male and female Siberian hamsters were gonadectomized and implanted with silastic capsules containing either cholesterol (C) or a mixture of estradiol (E) or testosterone (T). Microdialysis sampling within the mPOA was conducted after 8 weeks of exposure to either an LD or an SD photoperiod. Blood samples were analyzed for LH and PRL, while dialysis samples were analyzed for NE and its major metabolite 3-methoxy-4 hydroxyphenylglycol (MHPG). The results revealed significant suppression of LH and PRL by exposure to the SD photoperiod in both males and females. For LH, the steroid implants suppressed circulating hormone levels under both photoperiods, whereas for PRL, steroid treatment facilitated circulating levels. In contrast, there were no significant effects of photoperiod on NE or MHPG release for either males or females, but there was a significant decrease in extracellular levels of these neurochemicals in steroid-treated animals. These data suggest that photoperiodic modulation of GnRH neuronal function by NE is achieved largely through the indirect effects of photoperiod on circulating gonadal steroids. PMID- 11435760 TI - The fiscal and psychic costs of admissions. PMID- 11435761 TI - Self-efficacy and OSCE performance among second year medical students. AB - Competent performance requires not only requisite knowledge and skills but also beliefs of personal efficacy to use both effectively. This study examined the confidence of second-year medical students regarding their OSCE performance. Students completed an OSCE at the end of their second year of medical school; their performance was rated using checklists containing key items. Ten minutes prior to the OSCE, students completed a brief survey, which included 31 items sampling confidence in performing various clinical skills required in the OSCE. In addition, students assessed their preparedness and their anxiety, and reported the number of hours they studied in preparation for the OSCE. A total of 82 identifiable surveys (73 %) were used in this analysis. Students with high self efficacy were more likely to score above the mean OSCE performance compared to low self-rated students (71% versus 51%), however self-efficacy was not significantly correlated to OSCE performance. A causal path model was constructed to predict OSCE performance. Performance in the clinical skills and biomedical science curricula both were related to perceived anxiety, which was related to self-efficacy. Preparedness was predicated on self-efficacy and itself predicted performance. Knowledge also had a strong direct link to performance. Performance is more than having the requisite abilities. Performance was found to be the product of complex relationships between skills and knowledge, mediated by perceptions of anxiety, self-confidence and preparedness. The model illustrates the importance of realistic self-appraisal for competent performance. PMID- 11435762 TI - Predicting holistic ratings of written performance assessments from analytic scoring. AB - The use of experts to judge performance assessments is desirable because ratings of performances, carried out by experts in the content domain of the examination, are often considered to be the "gold standard." However, one drawback of using experts to rate performances is the high cost involved. A more economic alternative for scoring performance assessments entails using analytic scoring, which typically involves assigning points to individual traits present in the performance, and summing to arrive at a single score. This strategy is less costly, but may lack the richness of holistic scoring. This study investigates the use of regression-based techniques to predict expert judgments on a written performance task from a combination of analytic scores. Potentially, this will result in scores that approximate the richness of holistic ratings while maintaining the cost-effectiveness of analytic scoring. Results show that a substantial proportion of variance in expert judgments can be explained by the analytic scores, but that decisions based on actual expert judgments and the predicted expert judgments were not sufficiently consistent to warrant the substitution of one score for the other. PMID- 11435763 TI - Students' perceptions of assessment practices in a traditional medical curriculum. AB - This study examines advanced medical students' perceptions of assessment practices and their ways of studying for examinations as related to their approaches to learning. This study further validates a cluster model obtained in a previous study through medical students' interviews. In this cluster model students were divided into four groups on the basis of their approaches to learning. The subjects (N = 35) were advanced medical students who volunteered to be interviewed. The interviews focused on learning strategies, study behaviour and perceptions of the learning environment. The results indicated that there were no differences in students' perceptions of the examination procedures. Students in all four groups criticised quite similarly the examination practices. However, the results showed that students in the four groups reported different ways of preparing for examinations and, furthermore, had different views of the most functional ways to study for them. This study brought into light problems that arise in a traditional medical curriculum, particularly concerning traditional assessment practices. PMID- 11435764 TI - Testing a causal model for learning in a problem-based curriculum. AB - The aim of this study was to identify the relationship between elements that are important for the tutorial group process and the individual learning process in a problem-based curriculum. The variables under investigation were student generated learning issues, individual learning process, reporting in the tutorial group, and achievement.A questionnaire containing 22 items was developed. Data were collected in the first year (N = 195) of the Medical School of the Maastricht University in the Netherlands during the academic year 1997-1998. The data were analysed using a structural modelling approach. The results indicate that the model fitted the data well. The path coefficients were moderately high, particular between the explanation-oriented approach and the depth of the reporting in the tutorial group. High path coefficients were also found between the depth of the reporting and achievement. In sum, the model gives insight into how important variables are related and it is recommended that data should be collected to test the model repeatedly. PMID- 11435765 TI - Predicting mastery level on a large-scale standardized patient test: a comparison of case and instrument score-based models using discriminant function analysis. AB - Clinical skills assessments have traditionally been scored via experts' ratings of examinee performance. However, this approach to scoring may be impractical in a large-scale context due to logistical and cost considerations as well as the increased probability of rater error. The purpose of this investigation was therefore to identify, using discriminant analysis, weighted score-based models that maximize the accuracy with which mastery level can be estimated for examinees taking a nationally administered standardized patient test. Additionally, the accuracy with which the resulting classification functions can be applied to predict mastery level for a cross-validation sample of examinees was also examined. Results suggest that it might be feasible to implement an automated scoring procedure in a cost-effective manner while still retaining the important facets of the decision-making process of expert raters. Cost-benefit, test development and psychometric implications of these results are important and discussed in the full paper. PMID- 11435766 TI - Reliability and validity of admissions tools used to select students for the health professions. AB - The selection of students for the health professions is typically a very competitive multi-staged process that includes assessment of both cognitive abilities and personal qualities. The need for reliable and valid assessment measures is obvious. This review of the health professions literature examines the evidence to support the use of various selection tools. It is clear that pre admission overall grade point average (GPA) is the best predictor of academic performance in all of the health professions; however, the relationship between pre-admission GPA and clinical performance is less clear. The Medical College Admission Test is a good predictor of performance of medical students in terms of in-course grades and licencing examination scores but a similar test does not exist in the other health professions. Controversy remains as to the value of personal interviews and written submissions as selection tools, although it is clear that training of assessors and explicit rating guidelines enhance their reliability and validity. Ongoing research is needed to find more reliable and valid ways of assessing non-cognitive characteristics of applicants. PMID- 11435788 TI - Stroke genomics: approaches to identify, validate, and understand ischemic stroke gene expression. AB - Sequencing of the human genome is nearing completion and biologists, molecular biologists, and bioinformatics specialists have teamed up to develop global genomic technologies to help decipher the complex nature of pathophysiologic gene function. This review will focus on differential gene expression in ischemic stroke. It will discuss inheritance in the broader stroke population, how experimental models of spontaneous stroke might be applied to humans to identify chromosomal loci of increased risk and ischemic sensitivity, and also how the gene expression induced by stroke is related to the poststroke processes of brain injury, repair, and recovery. In addition, we discuss and summarise the literature of experimental stroke genomics and compare several approaches of differential gene expression analyzes. These include a comparison of representational difference analysis we have provided using an experimental stroke model that is representative of stroke evolution observed most often in man, and a summary of available data on stroke differential gene expression. Issues regarding validation of potential genes as stroke targets, the verification of message translation to protein products, the relevance of the expression of neuroprotective and neurodestructive genes and their specific timings, and the emerging problems of handling novel genes that may be discovered during differential gene expression analyses will also be addressed. PMID- 11435789 TI - Uncoupling of cerebral blood flow and metabolism after cerebral contusion in the rat. AB - Positron emission tomography scans of patients with head injuries often show discrete areas of increased 18F-fluorodeoxyglucose uptake ("hot spots") when performed hours to days after the initial ictus. Using quantitative autoradiographic methods, the authors have investigated whether cerebral blood flow and glucose metabolism are uncoupled 2 hours after controlled head injury in an animal model, and whether any "hot spots" are accompanied by changes in cerebral glucose concentration. Experiments were performed on 18 anesthetized, ventilated (1.5% halothane in 2:1 nitrous oxide:oxygen) Sprague-Dawley rats weighing 300 to 330 g. A burr hole was made over the left parietal cortex, and all animals received a piston impact on the intact dura (2 mm in diameter, 2.0 m/sec, 2 mm in depth). All animals remained anesthetized and ventilated for a further 2 hours, after which quantitative autoradiography was used to determine either (1) local cerebral blood flow (LCBF) using 14C-iodoantipyrine, (2) local cerebral glucose utilization (LCGU) using 14C-deoxyglucose, or (3) local cerebral glucose content (LCGC) using 14C-methylglucose. Local CBF, LCGU, and LCGC were measured in five regions adjacent to the contusion, and values then were normalized on the contralateral cortex. Normalized LCBF, LCGU, or LCGC varied in parallel in ipsilateral cortex (no change) and in the ischemic core of the contusion (reduced). However, there were marked changes in the patterns observed in the boundary zone (within 1 mm of the contusion). In all six rats used for LCGU measurement, there were discrete areas of high metabolism, whereas in all six rats used for LCBF measurement, flow was universally depressed in the boundary zone. Of the six rats used for LCGC determination, there was a discrete area of high signal in only one. The authors conclude that there are discrete areas of uncoupling of cerebral blood flow and metabolism after head injury within 2 hours of cerebral contusion in the rat that cannot be explained by changes in cerebral glucose content in the majority of animals. PMID- 11435790 TI - Anatomic and biochemical correlates of the dopamine transporter ligand 11C-PE2I in normal and parkinsonian primates: comparison with 6-[18F]fluoro-L-dopa. AB - Positron emission tomography (PET) coupled to 6-[18F]Fluoro-L-Dopa (18F-Dopa) remains the gold standard for assessing dysfunctionality concerning the dopaminergic nigrostriatal pathway in Parkinson's disease and related disorders. The use of ligands of the dopamine transporters (DAT) is an attractive alternative target; consequently, the current aim was to validate one of them, 11C-PE2I, using a multiinjection modeling approach allowing accurate quantitation of DAT densities in the striatum. Experiments were performed in three controls, three MPTP-treated (parkinsonian) baboons, and one reserpine-treated baboon. 11C PE2I B'max values obtained with this approach were compared with 18F-Dopa input rate constant values (Ki), in vitro Bmax binding of 125I-PE2I, and the number of dopaminergic neurons in the substantia nigra estimated postmortem by stereology. In the caudate nucleus and putamen, control values for 11C-PE2I B'max were 673 and 658 pmol/mL, respectively, whereas it was strongly reduced in the MPTP treated (B'max = 26 and 36 pmol/mL) and reserpine-treated animals (B'max = 338 and 483 pmol/mL). In vivo 11C-PE2I B'max values correlated with 18F-Dopa Ki values and in vitro 125I-PE2I Bmax values in the striatum and with the number of nigral dopaminergic neurons. Altogether, these data support the use of 11C-PE2I for monitoring striatal dopaminergic disorders and the effect of potential neuroprotective strategies. PMID- 11435791 TI - Quantitative comparison of the bolus and steady-state methods for measurement of cerebral perfusion and oxygen metabolism: positron emission tomography study using 15O-gas and water. AB - To evaluate a new simplified bolus method for measurement of cerebral perfusion and metabolism, the parametric images with that method were compared with those obtained from the conventional steady-state method with 15O-gas. The new method also provided images of arterial blood volume (V0), which is a different parameter from cerebral blood volume (CBV) obtained using a C15O technique. Seven healthy volunteers and 10 patients with occlusive cerebrovascular diseases underwent positron emission tomography (PET) scans with both methods. Three weighted integration was applied to calculate regional cerebral blood flow (rCBF) and regional cerebral metabolic rate of oxygen (rCMRO2) in the bolus method. Global and regional CBF and CMRO2 in volunteers were compared between the two methods and used as control data. Regional values in patients also were evaluated to observe differences between the bilateral hemispheres. Both rCBF and rCMRO2 were linearly well correlated between the two methods, although global difference in CMRO2 was significant. The difference in each parametric image except for V0 was significant between the bilateral hemispheres in patients. The bolus method can simplify oxygen metabolism studies and yield parametric images comparable with those with the steady-state method, and can allow for evaluation of V0 simultaneously. Increase in CBV without a change in V0 suggested the increase might mainly be caused by venous dilatation in the ischemic regions. PMID- 11435792 TI - Hypoperfusion without ischemia surrounding acute intracerebral hemorrhage. AB - A zone of hypoperfusion surrounding acute intracerebral hemorrhage (ICH) has been interpreted as regional ischemia. To determine if ischemia is present in the periclot area, the authors measured cerebral blood flow (CBF), cerebral metabolic rate of oxygen (CMRO2), and oxygen extraction fraction (OEF) with positron emission tomography (PET) in 19 patients 5 to 22 hours after hemorrhage onset. Periclot CBF, CMRO2, and OEF were determined in a 1-cm-wide area around the clot. In the 16 patients without midline shift, periclot data were compared with mirror contralateral regions. All PET images were masked to exclude noncerebral structures, and all PET measurements were corrected for partial volume effect due to clot and ventricles. Both periclot CBF and CMRO2 were significantly reduced compared with contralateral values (CBF: 20.9 +/- 7.6 vs. 37.0 +/- 13.9 mL 100 g( 1) min(-1), P = 0.0004; CMRO2: 1.4 +/- 0.5 vs. 2.9 +/- 0.9 mL 100 g(-1) min(-1), P = 0.00001). Periclot OEF was less than both hemispheric OEF (0.42 +/- 0.15 vs. 0.47 +/- 0.13, P = 0.05; n = 19) and contralateral regional OEF (0.44 +/- 0.16 vs. 0.51 +/- 0.13, P = 0.05; n = 16). In conclusion, CMRO2 was reduced to a greater degree than CBF in the periclot region in acute ICH, resulting in reduced OEF rather than the increased OEF that occurs in ischemia. Thus, the authors found no evidence for ischemia in the periclot zone of hypoperfusion in acute ICH patients studied 5 to 22 hours after hemorrhage onset. PMID- 11435793 TI - Induction of the HSP110/105 family in the rat hippocampus in cerebral ischemia and ischemic tolerance. AB - Recently, the authors isolated a novel gene of the HSP110 family, ischemia responsive protein 94 kDa (irp94), and demonstrated the expression of this gene after transient forebrain ischemia. In the current study, the authors investigated the expression profiles of all HSP110 family members including hsp110/105 and osp94/apg-1, after transient forebrain ischemia using rat four vessel occlusion model. Among three members of the HSP110 family, induction of hsp110/105 was the most prominent after ischemia. hsp110/105 mRNA expression was clearly enhanced from 4 to 24 hours after a 6-minute or longer ischemic period. First, hsp110/105 mRNA expression was induced in the dentate gyrus, and later in the pyramidal layer. HSP110/105 protein expression also was enhanced by a 6 minute or longer period of ischemia. Profiles of HSP110/105 expression after ischemia were similar to those of inducible HSP70. After transient forebrain ischemia for 10 minutes, HSP110/105 protein was induced in the dentate gyrus and the CA3 pyramidal layer, but not in the CA1 pyramidal neurons. However, 6 minutes of ischemia induced the HSP110/105 protein, as well as the HSP70 protein, in the CA1 region. CA1 pyramidal neurons expressing HSP110/105 acquired tolerance against subsequent severe ischemia. In conclusion, HSP110/105 showed the most prominent induction after ischemia among the three HSP110 gene family members. Colocalization of HSP110/105 and HSP70 in the CA1 neurons that acquired tolerance suggested that induced HSP110/105 might contribute to ischemic tolerance together with HSP70. PMID- 11435794 TI - Increased expression of transforming growth factor-beta after cerebral ischemia in the baboon: an endogenous marker of neuronal stress? AB - There has been an increasing interest in recent years in the evaluation of the neuronal and glial responses to ischemic insult. Some cytokines, including transforming growth factor-beta (TGF-beta), that are overexpressed after experimental stroke in rodents are thought to be implicated in the neuronal processes that lead to necrosis. Thus, such cytokines could predict tissue fate after stroke in humans, although data are currently sparse for gyrencephalic species. The current study addressed the expression pattern of TGF-beta1 in a nonhuman primate model of middle cerebral artery occlusion. Focal permanent ischemia was induced for 1 or 7 days in 6 baboons and the following investigations were undertaken: cerebral oxygen metabolism (CMRO2) positron emission tomography studies, magnetic resonance imaging, postmortem histology, and reverse transcription-polymerase chain reaction. The aim of the current study was to correlate the expression of TGF-beta1 to the underlying metabolic and histologic state of the threatened cerebral parenchyma. The authors evidenced increased TGF-beta1 mRNA levels (up to 25-fold) in those regions displaying a moderate (20% to 49%) reduction in CMRO2. The current findings suggest that the greatly enhanced expression of TGF-beta1 in the penumbral zones that surround tissue destined to infarction may represent a robust index of potentially salvageable brain. The current investigation, in the nonhuman primate, strengthens the authors' hypothesis, derived from rodent models, that TGF-beta1 may be involved in the physiopathology of human stroke. PMID- 11435795 TI - Chronic cerebral hypoperfusion induces MMP-2 but not MMP-9 expression in the microglia and vascular endothelium of white matter. AB - White matter lesions are closely associated with cognitive impairment and motor dysfunction in the aged. To explore the pathophysiology of these lesions, the authors examined the expression of matrix metalloproteinase-2 (MMP-2) and MMP-9 in the white matter in a rat model of chronic cerebral hypoperfusion. After bilateral clipping of the common carotid arteries, myelin staining revealed demyelinating changes in the optic tract and the corpus callosum on day 7. Zymographic analyses indicated an increase in the level of MMP-2, but not MMP-9, after the hypoperfusion. Immunohistochemical analyses revealed the presence (most abundantly on day 3) of MMP-2-expressing activated microglia in the optic tract and corpus callosum. In contrast, the capillary endothelial cells expressed MMP-2 later. IgM-immunoreactive glial cells were absent in the sham-operated animals, but were present in the hypoperfused animals by day 3, reflecting the disrupted blood-brain barrier. These findings suggest that the main sources of the elevated MMP-2 were the microglia and the endothelium, and that these cells may contribute to the remodeling of the white matter myelin and microvascular beds in chronic cerebral hypoperfusion. PMID- 11435796 TI - Rapid loss of microvascular integrin expression during focal brain ischemia reflects neuron injury. AB - The integrity of cerebral microvessels requires the close apposition of the endothelium to the astrocyte endfeet. Integrins alpha1beta1 and alpha6beta4 are cellular matrix receptors that may contribute to cerebral microvascular integrity. It has been hypothesized that focal ischemia alters integrin expression in a characteristic time-dependent manner consistent with neuron injury. The effects of middle cerebral artery occlusion (MCAO) and various periods of reperfusion on microvasclar integrin alpha1beta1 and alpha6beta4 expression were examined in the basal ganglia of 17 primates. Integrin subunits alpha1 and beta1 colocalized with the endothelial cell antigen CD31 in nonischemic microvessels and with glial fibrillary acidic protein on astrocyte fibers. Rapid, simultaneous, and significant disappearance of both integrin alpha1 and beta1 subunits and integrin alpha6beta4 occurred by 2 hours MCAO, which was greatest in the region of neuron injury (ischemic core, Ic), and progressively less in the peripheral (Ip) and nonischemic regions (N). Transcription of subunit beta1 mRNA on microvessels increased significantly in the Ic/Ip border and in multiple circular subregions within Ic. Microvascular integrin alpha1beta1 and integrin alpha6beta4 expression are rapidly and coordinately lost in Ic after MCAO. With loss of integrin alpha1beta1, multiple regions of microvascular beta1 mRNA up-regulation within Ic suggest that microvessel responses to focal ischemia are dynamic, and that multiple cores, not a single core, are generated. These changes imply that microvascular integrity is modified in a heterogeneous, but ordered pattern. PMID- 11435797 TI - Delayed hemorrhagic hypotension exacerbates the hemodynamic and histopathologic consequences of traumatic brain injury in rats. AB - Alterations in cerebral autoregulation and cerebrovascular reactivity after traumatic brain injury (TBI) may increase the susceptibility of the brain to secondary insults, including arterial hypotension. The purpose of this study was to evaluate the consequences of mild hemorrhagic hypotension on hemodynamic and histopathologic outcome after TBI. Intubated, anesthetized male rats were subjected to moderate (1.94 to 2.18 atm) parasagittal fluid-percussion (FP) brain injury. After TBI, animals were exposed to either normotension (group 1: TBI alone, n = 6) or hypotension (group 2: TBI + hypotension, n = 6). Moderate hypotension (60 mm Hg/30 min) was induced 5 minutes after TBI or sham procedures by hemorrhage. Sham-operated controls (group 3, n = 7) underwent an induced hypotensive period, whereas normotensive controls (group 4, n = 4) did not. For measuring regional cerebral blood flow (rCBF), radiolabeled microspheres were injected before, 20 minutes after, and 60 minutes after TBI (n = 23). For quantitative histopathologic evaluation, separate groups of animals were perfusion-fixed 3 days after TBI (n = 22). At 20 minutes after TBI, rCBF was bilaterally reduced by 57% +/- 6% and 48% +/- 11% in cortical and subcortical brain regions, respectively, under normotensive conditions. Compared with normotensive TBI rats, hemodynamic depression was significantly greater with induced hypotension in the histopathologically vulnerable (P1) posterior parietal cortex (P < 0.01). Secondary hypotension also increased contusion area at specific bregma levels compared with normotensive TBI rats (P < 0.05), as well as overall contusion volume (0.96 +/- 0.46 mm(3) vs. 2.02 +/- 0.51 mm(3), mean +/- SD, P < 0.05). These findings demonstrate that mild hemorrhagic hypotension after FP injury worsens local histopathologic outcome, possibly through vascular mechanisms. PMID- 11435798 TI - Increased cerebral infarct volumes in polyglobulic mice overexpressing erythropoietin. AB - There is increasing evidence that erythropoietin (Epo) has a protective function in cerebral ischemia. When used for treatment, high Epo plasma levels associated with increases in blood viscosity, however, may counteract beneficial effects of Epo in brain ischemia. The authors generated two transgenic mouse lines that overexpress human Epo preferentially, but not exclusively, in neuronal cells. In mouse line tg21, a fourfold increase of Epo protein level was found in brain only, whereas line tg6 showed a dramatic increase of cerebral and systemic transgene expression resulting in hematocrit levels of 80%. Cerebral blood flow (CBF), as determined by bolus tracking magnetic resonance imaging, was not altered in the tg6 line. The time-to-peak interval for the tracer, however, increased approximately threefold in polyglobulic tg6 mice. Immunohistochemical analysis revealed an increase in dilated vessels in tg6 mice, providing an explanation for unaltered CBF in polyglobulic animals. Permanent occlusion of the middle cerebral artery (pMCAO) led to similar perfusion deficits in wild-type, tg6, and tg21 mice. Compared with wild-type controls, infarct volumes were not significantly smaller (22%) in tg21 animals 24 hours after pMCAO, but were 49% enlarged (P < 0.05) in polyglobulic tg6 mice. In the latter animals, elevated numbers of Mac-1 immunoreactive cells in infarcted tissue suggested that leukocyte infiltration contributed to enlarged infarct volume. The current results indicate that moderately increased brain levels of Epo in tg21 transgenic mice were not sufficient to provide significant tissue protection after pMCAO. The results with tg6 mice indicate that systemic chronic treatment with Epo associated with elevated hematocrit might deteriorate outcome after stroke either because of the elevated hematocrit or other chronic effects. PMID- 11435799 TI - Protein aggregation after focal brain ischemia and reperfusion. AB - Two hours of transient focal brain ischemia causes acute neuronal death in the striatal core region and a somewhat more delayed type of neuronal death in neocortex. The objective of the current study was to investigate protein aggregation and neuronal death after focal brain ischemia in rats. Brain ischemia was induced by 2 hours of middle cerebral artery occlusion. Protein aggregation was analyzed by electron microscopy, laser-scanning confocal microscopy, and Western blotting. Two hours of focal brain ischemia induced protein aggregation in ischemic neocortical neurons at 1 hour of reperfusion, and protein aggregation persisted until neuronal death at 24 hours of reperfusion. Protein aggregates were found in the neuronal soma, dendrites, and axons, and they were associated with intracellular membranous structures during the postischemic phase. High resolution confocal microscopy showed that clumped protein aggregates surrounding nuclei and along dendrites were formed after brain ischemia. On Western blots, ubiquitinated proteins (ubi-proteins) were dramatically increased in neocortical tissues in the postischemic phase. The ubi-proteins were Triton-insoluble, indicating that they might be irreversibly aggregated. The formation of ubi protein aggregates after ischemia correlated well with the observed decrease in free ubiquitin and neuronal death. The authors concluded that proteins are severely damaged and aggregated in neurons after focal ischemia. The authors propose that protein damage or aggregation may contribute to ischemic neuronal death. PMID- 11435800 TI - Hydroxyfasudil, an active metabolite of fasudil hydrochloride, relaxes the rabbit basilar artery by disinhibition of myosin light chain phosphatase. AB - Fasudil hydrochloride (AT877, hexahydro-1-(5-isoquinolinesulfonyl)-1H-1,4 diazepine hydrochloride, identical to HA1077) inhibits cerebral vasospasm after subarachnoid hemorrhage in experimental animals and humans. In the current study, the vasorelaxing mechanism of hydroxyfasudil, a hydroxylated metabolite of fasudil hydrochloride, was determined in the rabbit basilar artery. The effects of hydroxyfasudil on tension, intracellular Ca2+ concentration ([Ca2+]i), and phosphorylation of the myosin light chain were examined using the isolated and intact or permeabilized rabbit basilar artery without endothelium in vitro. In the intact rabbit basilar artery, hydroxyfasudil elicited a concentration dependent relaxation of the artery precontracted with 1 nmol/L endothelin-1 (ET 1) plus 20 mmol/L KCl without any significant decrease in [Ca2+]i as determined by fura-2 microfluorometry (IC50: 5.1 +/- 4.6 micromol/L). The relaxation induced by hydroxyfasudil was accompanied with dephosphorylation of the myosin light chain. In the permeabilized preparation, hydroxyfasudil inhibited the contraction induced by ET-1, guanosine 5'-O-(3-thiotriphosphate), or the catalytic subunit of rho-associated kinase, but it did not inhibit Ca2+-induced contraction under the condition of inhibited myosin light chain phosphatase. Hydroxyfasudil showed a greater relaxant effect under decreased adenosine triphosphate (ATP) levels. The present study indicated that hydroxyfasudil relaxes the rabbit basilar artery mainly by disinhibiting myosin light chain phosphatase through the inhibition of rho-associated kinase and that this effect depends on the intracellular ATP concentration. PMID- 11435801 TI - Sleep disorders in neurology. PMID- 11435802 TI - Narcolepsy: clinical features, new pathophysiologic insights, and future perspectives. AB - Narcolepsy is characterized by excessive daytime sleepiness and abnormal manifestations of rapid eye movement sleep such as cataplexy. The authors review the clinical features of narcolepsy, including epidemiology, symptoms, diagnosis, and treatment, in detail. Recent findings show that a loss of hypocretin producing neurons lies at the root of the signs and symptoms of narcolepsy. The authors review the current state of knowledge on hypocretin anatomy, physiology, and function with special emphasis on the research regarding the hypocretin deficiency in narcolepsy, which may also explain associated features of the disorder, such as obesity. Lastly, they discuss some future perspectives for research into the pathophysiology of sleep/wake disorders, and the potential impact of the established hypocretin deficiency on the diagnosis and treatment of narcolepsy. PMID- 11435803 TI - Interactions between sleep and epilepsy. AB - Sleep is one of the best-documented factors influencing the expression of seizures and interictal discharges. Janz studied the relation between seizures and the sleep/wake cycle and divided the epilepsies into three categories: nocturnal, awakening, and diffuse. Since then, the effect of sleep on the ictal and interictal manifestations of epilepsy has been studied extensively. Many seizures are activated by sleep or arousal from sleep. Interictal discharges are also seen more commonly during sleep, with the greatest activation seen during nonrapid eye movement sleep. Sleep not only increases the frequency of epileptiform abnormalities, but also may alter their morphology and distribution. Sleep deprivation also facilitates both epileptiform abnormalities and seizures. Seizures, on the other hand, also impact sleep. Epileptic patients demonstrate multiple sleep abnormalities, including an increased sleep latency, fragmented sleep, increased awakenings and stage shifts, and an increase in stages 1 and 2 of nonrapid eye movement sleep. These disturbances may in turn be modulated by antiepileptic treatment. This review summarizes the interactions between sleep and epilepsy, including the timing of seizures during the sleep/wake cycle, the influence of sleep on various seizure disorders, the effects of sleep deprivation, and the changes in sleep patterns caused by seizures and their treatment. PMID- 11435804 TI - Restless legs syndrome: a review of clinical and pathophysiologic features. AB - Restless legs syndrome (RLS), although long ignored and still much underdiagnosed, disrupts the life and sleep considerably of those who have it. Recent clinical and basic research provides for better definition and pathophysiologic understanding of the disorder. The body of knowledge about this disorder has been expanding rapidly during the past decade and it has altered our concepts of this disorder. This review of RLS covers history, diagnosis, morbidity of sleep disturbance, relation to periodic limb movements in both sleep and waking, secondary causes, severity assessment methods, phenotypes for possible genetic patterns, epidemiology, pathophysiology, and medical treatment considerations. The emphasis on pathophysiology includes consideration of central nervous system localization, neurotransmitter and other systems involved, and the role of iron metabolism. Studies to date support the authors' recently advanced iron-dopamine model of RLS. PMID- 11435805 TI - Rem sleep behavior disorder: potential relationship to post-traumatic stress disorder. AB - Rapid eye movement sleep behavior disorder (RBD) is a parasomnia in which there is enactment, often violent, of dream mentation. Although this syndrome is sometimes associated with neurologic disorders, psychiatric comorbidity is not typical. The authors present a unique series of veterans with RBD. A high incidence of comorbidity with post-traumatic stress disorder is noted. The literature on RBD is reviewed, and the coexistence of RBD and post-traumatic stress disorder is reasoned. The authors suggest that it is possible that similar neuropathologic processes are responsible for both conditions, at times in the same patient. PMID- 11435806 TI - Success of ambulatory EEG in children. AB - SUMMARY: Continuous ambulatory EEG (AEEG) monitoring is a method used to (1) determine seizure type and location of seizure onset, and (2) to discriminate between epileptic and nonepileptic events. This study was performed to determine how successful AEEG would be in recording seizures when the events were reported to occur at least 3 days per week. AEEGs of children who were patients at Children's Hospital Oakland between December 1993 and June 1997 were reviewed to see why the recordings were performed and to determine whether typical seizures were recorded. Children who had seizurelike events needed to have typical spells 3 days or more per week to justify obtaining AEEG. Most AEEGs were performed to discriminate between epileptic and nonepileptic seizures. A total of 167 children underwent AEEG recording. Ten were recorded to determine whether they were having frequent subtle seizures or frequent interictal epileptiform discharges. The remaining 157 patients had discrete events. A total of 140 children (89%) had their typical spells recorded. A total of 107 of these children (76%) had nonepileptic events. Average duration of recording was 1.9 days. AEEG is very successful in recording children's seizurelike events when parents report events occur at least 3 days per week. The procedure is well tolerated and there are few technical problems that prolong recording time. PMID- 11435807 TI - Current practice in administration and clinical criteria of emergent EEG. AB - Policies of administration and availability of EEG offered during nonbusiness hours vary widely among EEG laboratories. The authors surveyed medical directors of accredited EEG laboratories (n = 84) to determine the ranges of availability and clinical indications for approval of continuously available emergent EEG (E EEG). Of 46 respondents, 37 (80%) offered E-EEG. Two centers recently lost funding for E-EEG. Availability was not associated with the total number of EEGs performed annually. The mean estimated response time from request to expert interpretation was 3 +/- 4 hours (range, 1-24 hours). The five clinical indications for which most respondents approved E-EEGs were possible nonconvulsive status epilepticus (100%), treatment of status epilepticus (84%), cerebral death exam (81%), diagnosis of convulsive status epilepticus (79%), and diagnosis of coma or encephalopathy (70%). Respondents disagreed widely when asked which clinical situations merited E-EEG, with some approving all requests and others denying all except for nonconvulsive status epilepticus. The wide range of current practice suggests that research focused on outcomes of aggressive, EEG-aided patient evaluation and treatment are needed to define better the costs and benefits of a continuously available EEG service. PMID- 11435808 TI - Continuous EEG monitoring in a patient with massive carbamazepine overdose. AB - The authors report a woman who took a massive overdose (OD) of carbamazepine (CBZ). On admission she was unconscious with absent brainstem reflexes and multifocal stimulus-sensitive myoclonus. Continuous EEG recordings showed a burst suppression pattern with bursts containing only generalized spikes accompanying myoclonic activity. Myoclonus and EEG bursts were both spontaneous and stimulus induced. With treatment, the serum CBZ concentration declined, and the EEG became more continuous and rhythmic without epileptiform discharges. Unfortunately, the patient died from adult respiratory distress syndrome. Autopsy revealed that cortical and subcortical structures were normal without neuronal necrosis or eosinophilia. Massive CBZ OD may produce a reversible encephalopathy that includes cortical hyperexcitability, a profound burst-suppression EEG pattern, and cranial nerve areflexia. Continuous EEG monitoring is helpful in managing seizures that occur as a complication of CBZ OD, after the course of recovery or worsening, and in providing assistance with prognosis. PMID- 11435809 TI - The development of spectral EEG changes during short periods of circulatory arrest. AB - The EEG was monitored in 56 patients during implantation of an internal cardioverter defibrillator. The purpose of this study was to determine the main EEG frequency ranges that represent ischemic changes during short periods of circulatory arrest. The EEG was recorded with a 16-channel common reference montage (Cz). After onset of circulatory arrest, the log spectral changes of three-epoch moving averages were calculated relative to the baseline spectrum. For factor analysis, 17 EEG periods were selected that showed changes progressing to an isoelectrical period. Topographic differences and the time course of quantitative EEG (qEEG) changes were studied in all 56 patients. For each patient the EEG period with the longest duration of circulatory arrest was chosen. Factor analysis revealed four factors that represented the spectral EEG changes occurring during circulatory arrest and recovery. The frequency intervals of these factors were 0 to 0.5 Hz, 1.5 to 3 Hz, 7.5 to 9.5 Hz, and 15 to 20 Hz for all channels. Only minor topographic differences were found in the power of the spectral changes; the sequence of events was similar for all electrode positions. The first EEG change after circulatory arrest was an initial increase in alpha power and a decrease in beta power. On average, after approximately 15 seconds alpha power started to decrease, beta power decreased further, delta-1 power started to increase, and delta-2 power started to decrease. After approximately 25 seconds, the delta-1 power increase appeared to plateau or to decrease. A circulatory arrest longer than approximately 30 seconds resulted in an isoelectrical EEG. After restoration of the circulation, there was a fast transient increase in delta-1 and delta-2 power, followed by a decrease to baseline. alpha and beta power showed a more gradual increase in power toward baseline and were the last to restore after 60 to 90 seconds. In general, the spectral changes in the alpha and beta frequency ranges were most pronounced and consistent. In conclusion, to detect intraoperative cerebral ischemia, monitoring of changes in the four frequency ranges found is preferable to monitoring changes in the classically defined frequency bands. Furthermore, these results stress the importance of the alpha and beta ranges in detecting cerebral ischemia. PMID- 11435810 TI - Correlation between cortical theta activity and hippocampal volumes in health, mild cognitive impairment, and mild dementia. AB - Cognitive decline is known to be associated with both increased theta power over frontal regions and hippocampal atrophy. The aim of this study was to reveal the relation between these parameters in groups with mild dementia, mild cognitive impairment, and healthy control subjects. The authors examined a preliminary randomly selected sample of 39 right-handed subjects joining the Leipzig Longitudinal Study of the Aged, consisting of 17 normal elderly subjects, 12 patients with mild cognitive impairment, and 10 patients with mild dementia assessed by Clinical Dementia Rating. All subjects were between 75 and 85 years old (mean age, 78 years; standard deviation, 2.78 years) and underwent EEG and brain MRI. Mean spectral power densities were calculated, and hippocampal body volume was measured. Significant negative linear correlations between theta power over frontal regions and hippocampal volumes were found. The results support the assumption about a relationship between hippocampal atrophy and theta power, and may be helpful for a better understanding of the course of Alzheimer's disease. PMID- 11435811 TI - Lower cervical origin of the P13-like potential in median SSEPS. AB - The authors studied the origin of the scalp P13-like potential in median somatosensory evoked potentials, which have been reported to be preserved in patients with cervicomedullary lesions or in brain death. There were five patients with high to middle cervical lesions (C2/3 or C3/4 level). Small P13 like potentials after P11 were identified for all patients with a noncephalic reference but not with an ear reference. Their onset latencies were slightly earlier than the expected latency of the true P13/14 onset. In two patients, delayed true P13/14s followed by N18s were identified with both noncephalic and ear references. The authors argue that the P13-like potential observed in these patients is a different entity from scalp P13 in normal subjects. Because the C3/4 vertebral level corresponds to the C5 cord level, the origin of the P13-like potential must be below C5, contradicting the previous opinion that it is generated at the cervicomedullary junction or at the high cervical dorsal column. The authors named this potential lower cervical P13 (or lcP13), and present an opinion that it is generated by the beginning of the second spinal ascending volley, which has been described by direct-recording studies in humans. PMID- 11435813 TI - Nephron sparing surgery for renal tumors: indications, techniques and outcomes. AB - PURPOSE: A contemporary review of the indications, techniques and outcomes is presented for nephron sparing approaches to solid renal masses, emphasizing their role for the treatment of renal cell carcinoma. We also reviewed the evolving role of minimally invasive forms of parenchymal sparing renal surgery. MATERIALS AND METHODS: MEDLINE and CANCERLIT computerized literature searches, and manual bibliographic reviews were performed to identify published peer reviewed articles pertaining to nephron sparing surgery or partial nephrectomy from 1980 to 2000. Pertinent articles were collated and reviewed. RESULTS: Nephron sparing surgery is increasingly being used to treat patients with solid renal lesions. The technical success rate of nephron sparing surgery is excellent, and operative morbidity and mortality are low. For renal cell carcinoma long-term cancer-free survival is comparable to that after radical nephrectomy, particularly for low stage disease. The overall incidence of local recurrence is low at 0% to 10%. For tumors 4 cm. or less local recurrence rates are even less at 0% to 3%. The risk of local recurrence depends primarily on the initial local pathological tumor stage. The reported incidence of multifocal renal cell carcinoma is approximately 15% and it also depends on tumor size, histology and stage. The risk of multifocal disease is low at less than 5% when the maximal diameter of the primary tumor is 4 cm. or less. Recent advances in renal imaging limit the radiographic evaluation necessary when planning complex nephron sparing approaches. Three-dimensional, volume rendered computerized tomography integrates all of the necessary information previously obtained by conventional computerized tomography, angiography, venography and pyelography into a single preoperative test, allowing better operative planning with maximal preservation of unaffected parenchyma in the remnant kidney. Minimally invasive modalities of tumor resection or destruction should be reserved for highly select patients and await improvements in technology, standardization of technique and long-term outcomes data before they may be completely integrated options. CONCLUSIONS: Nephron sparing surgery provides effective therapy for patients in whom preservation of renal function is a relevant clinical consideration. The importance of meticulous operative technique for achieving acceptable oncological and functional outcomes is emphasized. Accumulating data in appropriately select patients suggest a long term functional advantage gained by the maximal preservation of unaffected renal parenchyma without sacrificing cancer control. PMID- 11435814 TI - Pelvic lymph node metastases from bladder cancer: outcome in 83 patients after radical cystectomy and pelvic lymphadenectomy. AB - PURPOSE: We evaluate the outcome in patients with node positive bladder cancer with particular reference to the effect of individual characteristics of positive nodes on survival after meticulous pelvic lymphadenectomy at cystectomy. MATERIALS AND METHODS: This prospective analysis contains 452 cases of bladder cancer staged preoperatively as N0M0, managed with pelvic lymphadenectomy and cystectomy between 1984 and 1997. A total of 83 (18%) patients with histologically confirmed node positive disease are included in our study. RESULTS: The median overall survival of patients with positive nodes was 20 months. Median 5-year survival was 29%. Patients who survived were found with positive nodes at each site in the pelvis. The median survival of 57 patients with less than 5 positive nodes was 27 months, compared with 15 months for 26 with 5 nodes or more (log-rank test p = 0.0027). Median survival of 26 patients with no lymph node capsule perforation was 93 months, compared with 16 months for 57 with capsule perforation (p = 0.0004). The median survival of 18 patients with a maximum diameter of lymph node metastasis up to 0.5 cm. was 64 months, compared with 16 months for 65 with nodal metastasis greater than 0.5 cm. (p = 0.024). Contralateral positive nodes were found in 16 of 39 (41%) patients with unilateral bladder cancer. CONCLUSIONS: Long-term survival is possible with node positive bladder cancer. Those patients with few as well as smaller and, therefore, unsuspected nodal metastases, and those without lymph node capsule perforation have the best results after removal of pelvic metastatic nodal disease. Because patients who survive may be found regardless of the site of pelvic nodal metastases, meticulous bilateral pelvic lymphadenectomy is warranted in all patients at the time of attempted curative cystectomy for bladder cancer, particularly if there is no clinical evidence of nodal involvement. PMID- 11435815 TI - Multiple endocrine neoplasia type 1 and adrenal lesions. AB - PURPOSE: We investigated the relationship of long-term pancreatic hormone hypersecretion with adrenal lesions in patients with multiple endocrine neoplasia type 1 and in those with sporadic pancreatic endocrine tumors. MATERIALS AND METHODS: We assessed the prevalence of adrenal lesions in 20 patients with multiple endocrine neoplasia type 1 and in a control group of 12 with sporadic pancreatic endocrine tumors. We also performed genetic testing for germline mutations of MEN1, the multiple endocrine neoplasia type 1 gene. RESULTS: Adrenal lesions were common in multiple endocrine neoplasia type 1, accounting for 35% of cases. All adrenal lesions were nonfunctioning and benign. The relative risk of adrenal tumors was higher in patients with multiple endocrine neoplasia type 1 than in controls (p <0.05). No apparent relationship was observed of hormonal pattern or genotype with adrenal disease. CONCLUSIONS: Hormone hypersecretion by pancreatic endocrine tumors is not the primary cause of the development of adrenal lesions and the role of the MEN1 gene in adrenal tumorigenesis remains unclear. Adrenal lesions follow a benign course in most multiple endocrine neoplasia type 1 cases but careful morphological and functional followup is advisable. PMID- 11435816 TI - In vitro study of ultrasound based real-time tracking of renal stones for shock wave lithotripsy: part 1. AB - PURPOSE: We developed a real-time tracking system for renal stones that decreases the number of shock waves and treatment time of shock wave lithotripsy. MATERIALS AND METHODS: Ultrasound images were analyzed to identify the renal stones. A computer software module for ultrasound image processing was developed to monitor stone location instantaneously. Another computer software module controlled generator movement in real time for tracking the stone. We used 13 ultrasound stone trajectories recorded from patients to test the system in vitro. Two tests were established to verify tracking system reliability. One test focused on improvement in the coincidence ratio, which denotes the matching extent of the stone within the effective focal area. The other test focused on improvement in the efficiency ratio, that is a decrease in the number of shocks for stone fragmentation. For each recorded stone trajectory 2 experiments were done under tracking and nontracking conditions. RESULTS: The average coincidence and efficiency ratios plus or minus standard deviation were 79.6% +/- 9.8% and 45.0% +/- 12.7% without tracking, and 97.0% +/- 3.0% and 85.5 +/- 6.8% with tracking, respectively. All tests were statistically significant (paired t test p <0.01). CONCLUSIONS: An ultrasound based real-time tracking system proved to be significantly helpful for in vitro lithotripsy. It appears that the tracking system may greatly decrease the number of shocks and treatment time for renal stones. PMID- 11435817 TI - Long-term renal functional effects of shock wave lithotripsy, percutaneous nephrolithotomy and combination therapy: a comparative study of patients with solitary kidney. AB - PURPOSE: We compared the long-term impact on renal function after shock wave lithotripsy, percutaneous nephrolithotomy or the 2 techniques combined in patients with a solitary kidney. MATERIALS AND METHODS: A total of 45 women and 38 men 15 to 86 years old (mean age 56.1) with a solitary kidney were treated with shock wave lithotripsy (53), percutaneous nephrolithotomy (18) or the 2 techniques combined (12). Before and after treatment serum creatinine, blood pressure and the calculated glomerular filtration rate were determined, and raw and calculated data were compared by the Kruskal-Wallis, Fisher exact and Wilcoxon rank sum tests, and the Spearman correlation coefficient. Followup was 1 to 166.5 months (mean 53.0, median 46.9) overall and statistically equivalent in the 3 treatment arms. RESULTS: Treatment groups were comparable in regard to patient age, sex distribution, weight, blood pressure and pretreatment serum creatinine. There was no significant difference in any evaluated pretreatment or posttreatment parameters and no difference in the change in any parameter after treatment. Stratifying patients to pretreatment serum creatinine less or greater than 2 mg./dl. likewise revealed no significant difference in the impact on long term renal function. However, pretreatment serum creatinine positively and strongly correlated with a positive change in the glomerular filtration rate after therapy. CONCLUSIONS: In this study there was no evidence that any of these 3 treatment modalities resulted in the deterioration of renal function even at long-term followup. This finding implies that shock wave lithotripsy, percutaneous nephrolithotomy and the 2 therapies combined are equally efficacious for preserving renal function when performed in patients with a solitary kidney. PMID- 11435818 TI - Experience with retroperitoneal laparoscopic adrenalectomy in 115 procedures. AB - PURPOSE: Laparoscopic adrenalectomy has become an effective option for removing small adrenal tumors. We evaluated the retroperitoneal approach with regard to operative complications, morbidity and hospital stay. MATERIALS AND METHODS: Between January 1995 and March 2000 we performed a total of 115 laparoscopic adrenalectomies via the retroperitoneal approach, including 70 on the left and 45 on the right side, in 64 women and 42 men 17 to 74 years old (mean age 49.3) with adrenal neoplasms. Average adrenal tumor size was 31 mm. (range 10 to 65). All procedures required 4 trocars and mean operative time was 118 minutes (range 45 to 240). RESULTS: There were no patient deaths. The conversion rate to open surgery was 0.8% and estimated blood loss was 77 ml. (range 0 to 550). At a mean followup of 23.4 months, morbidity was 15.5% with intraoperative vascular injury in 3 cases (3.4%) and postoperative complications in 12.1%, including wound infection, deep hematoma, parietal dehiscence and severe pneumopathy. Average hospital stay was 4 days and mean duration of analgesic requirement was 2 days (range 1 to 5). CONCLUSIONS: The retroperitoneal approach to laparoscopic adrenalectomy appears to be minimally invasive and safe for adrenal tumors not larger than 5 cm. PMID- 11435819 TI - Laparoscopic bilateral hand assisted nephrectomy for autosomal dominant polycystic kidney disease: initial experience. AB - PURPOSE: The laparoscopic technique for bilateral nephrectomy in patients with autosomal dominant polycystic kidney disease is technically difficult. The procedure may be more acceptable if alterations to the technique made it safer and easier to perform. We describe our initial experience with, and the feasibility and potential benefits of hand assisted laparoscopic nephrectomy for approaching these large kidneys in patients with autosomal dominant polycystic kidney disease. MATERIALS AND METHODS: This approach was successfully applied in 3 patients with end stage renal disease due to autosomal dominant polycystic kidney disease. After obtaining transumbilical pneumoperitoneum ports were placed in the umbilicus (12 mm.), sub-xiphoid in the midline (12 mm.) and subcostal in the midclavicular line on each side (12 mm.). The table was tilted 40 degrees away from the planned side of initial nephrectomy with the patient in the half lateral position. A 7 cm. midline incision was made that incorporated the umbilical port and a commercially available hand assistance device was positioned. One surgeon hand was inserted into the abdomen to serve as a retractor/blunt dissector, while the other operated the electrosurgical instruments. The right hand was inserted for left nephrectomy and the left hand was inserted for right nephrectomy. The laparoscope was passed via the sub xiphoid port and the instruments were placed through the ipsilateral subcostal laparoscopic port. Nephrectomy was completed and the specimen was removed through the hand port incision by draining the cysts as they were exposed to view via the midline incision. When dissection was difficult, an additional port was placed in the anterior axillary line at the umbilical level. Some cysts were ruptured or aspirated to decrease overall kidney size and make extraction possible via the 6 to 7 cm. midline incision. RESULTS: All procedures were successfully completed. Mean operative time for bilateral hand assisted laparoscopic nephrectomy was 5.5 hours (range 4.5 to 6.6). Estimated blood loss was 200 cc or less. Patients resumed oral intake on postoperative day 1. The mean amount of parenteral analgesics required postoperatively was decreased. Mean hospital stay was 4.3 days but it was 3 days when considering nephrectomy only. Patients returned to normal activity after an average of 2 weeks. There was sustained resolution of preoperative discomfort based on pain analog scales. At 1 month or less all patients recorded absent pain. They uniformly noticed improved preoperative pulmonary and gastrointestinal symptoms CONCLUSIONS: Hand assisted laparoscopic nephrectomy in patients with autosomal dominant polycystic kidney disease makes bilateral nephrectomy a reasonable option. The bilateral procedure may be performed as rapidly as laparoscopic only, unilateral nephrectomy in these cases. The advantages of the hand assisted approach include using tactile sensation to facilitate dissection, rapid blunt finger dissection, hand retraction and the application of immediate tamponade when needed. This procedure provides the benefits of minimal intraoperative blood loss, minimal postoperative pain, brief hospital stay and rapid convalescence in this group of patients at high risk. PMID- 11435820 TI - Extraperitoneal laparoscopic pyeloplasty: a multicenter study of 55 procedures. AB - PURPOSE: We assessed the feasibility, reproducibility and morbidity of retroperitoneal laparoscopic pyeloplasty for ureteropelvic junction obstruction. MATERIALS AND METHODS: A total of 55 retroperitoneal laparoscopic pyeloplasties were performed at 3 institutions between September 1996 and May 2000 in 33 women and 21 men. Results were analyzed in regard to radiological assessment by excretory urography at 3 months, complications and hospital stay. RESULTS: We performed dismembered pyeloplasty in 48 cases and Fenger plasty in 7 cases. Crossing vessels were noted in 23 patients. The conversion rate was 5.4%. Mean operative time was 185 minutes (range 100 to 260), mean hospital stay was 4.5 days (range 1 to 14) and mean followup was 14.4 months (range 6 to 43.6). The overall complication rate was 12.7%. Complications in 7 patients included hematoma in 3, urinoma in 1, severe pyelonephritis in 1 and anastomotic stricture in 2 requiring open pyeloplasty at 3 weeks and delayed balloon incision at 13 months, respectively. Excretory urography in 50 patients and ultrasound in 4 showed decreased hydronephrosis in 88.9% at 3 months. Normal physical activity and absent pain were reported by 47 patients (87%) 1 month after surgery. CONCLUSIONS: Retroperitoneal laparoscopic pyeloplasty seems to be a valuable alternative to open pyeloplasty for ureteropelvic junction obstruction. The long term outcome must be assessed before this procedure may be definitively validated. PMID- 11435821 TI - The laparoscopic Boari flap. AB - PURPOSE: Laparoscopic management of ureteral pathologies, such as ureteropelvic junction obstruction, ureteral calculi and retroperitoneal fibrosis, has proved to be highly effective. We present our initial experience with the laparoscopic Boari flap in regard to feasibility, safety and short-term results. MATERIALS AND METHODS: Three patients who presented with distal ureteral obstruction underwent preoperative radiographic evaluation, including excretory urography, computerized tomography and retrograde pyelography, that showed upper urinary tract dilatation at the site of obstruction. The contralateral upper urinary tract was normal in all cases. Ureteroureterostomy and ureteroneocystostomy was not feasible because of stricture length and a laparoscopic Boari flap procedure was performed. Renal function, symptom improvement and radiological studies, including excretory urography and cystography, were assessed 3 and 6 months postoperatively. RESULTS: All procedures were successfully performed without any intraoperative complications or need for open conversion. Excretory urography showed good drainage with no obstruction of urine flow and all patients had grade I vesicoureteral reflux on cystography. CONCLUSIONS: The laparoscopic Boari flap is feasible using currently available laparoscopic suturing techniques. Longer followup and larger series of patients are necessary to provide data comparable to that of the open approach. PMID- 11435822 TI - Reevaluation of the 1997 TNM classification for renal cell carcinoma: T1 and T2 cutoff point at 4.5 rather than 7 cm. better correlates with clinical outcome. AB - PURPOSE: We analyzed the effects of the change in TNM classification from the 1987 to the 1997 version and suggest a modified tumor size cutoff point between T stages 1 and 2 for renal cell carcinoma. MATERIALS AND METHODS: We evaluated a database containing the records of 661 patients who underwent nephrectomy between 1989 and 1999. The effect of the change in TNM classification on the distribution of patients between stages, the rates of M+ and N+ disease, and the local and distant recurrence rates were outlined for 280 patients with T stages 1 and 2 disease. The Cox model was used to identify the optimal cutoff point between T1 and T2 disease, and the resulting effect of adopting this cutoff was outlined. RESULTS: A total of 174 and 128 cases were down staged from 1987 version stage T2 to 1997 version stage T1 and from 1987 TNM stage II to 1997 TNM stage I, respectively. Survival was not significantly different in patients with 1997 TNM stages I and II disease due to a lack of survival difference during the first 2 years of followup. Stage shift also caused an increase in average tumor size, the proportion of patients with high grade cancer, and M+ and N+ disease at diagnosis in 1997 stages T1 and T2 as well as an increase in the proportion of 1997 stage T2N0M0 cases at diagnosis with systemic failure. Analysis of 11 potential cutoff points between 1 and 10 cm. revealed that 4.5 cm. was most predictive of patients survival (hazards ratio 4.99, p = 0.0001). Using this cutoff resulted in improved discriminatory power of the TNM classification and a moderating effect on the distribution of patients, average tumor size, high grade disease, M+ and N+ disease at diagnosis, and systemic failure between T(14.5) and T(24.5) compared with 1997 T1 and T2. CONCLUSIONS: Our data imply that the current cutoff point of 7 cm. between stages T1 and T2 tumors is too high. Lowering the cutoff to 4.5 cm. resulted in better discriminatory power of the TNM classification in our dataset. This observation should be further validated by external data. PMID- 11435823 TI - Adrenal sparing surgery during radical nephrectomy in patients with renal cell cancer: a new algorithm. AB - PURPOSE: Ipsilateral adrenalectomy is usually performed during radical nephrectomy because of renal cell cancer. Because renal tumors are detected more often in the earlier stages due to widespread use of ultrasound and computerized tomography, we define a subset of patients who would be eligible for adrenal sparing surgery. In a retrospective analysis we evaluated whether parameters obtained preoperatively are able to predict adrenal metastasis. MATERIALS AND METHODS: A total of 866 consecutive patients who underwent nephrectomy and ipsilateral adrenalectomy from 1983 to 1999 were evaluated. Preoperative parameters, including tumor size, location, clinical stage, number of tumors, and patient age and sex, were retrospectively compared with the histological results. Univariate and multivariate analyses were performed. RESULTS: A total of 27 (3.1%) adrenal metastases were noted in the 866 patients, and 63% were on the left side and 37% on the right side. Mean tumor size was 10 cm. with versus 6 cm. without adrenal involvement. Of the 27 patients 21 had multiple metastases at diagnosis and only 6 (0.7% of all 866) presented with solitary ipsilateral adrenal metastasis. Univariate and multivariate analyses revealed tumor size and M stage as best preoperative predictors of adrenal involvement. CONCLUSIONS: Adrenal sparing surgery is possible, and we suggest a new algorithm. If maximum tumor size measured by computerized tomography is less than 8 cm. and staging examination does not show organ or lymph node metastases, adrenalectomy is not necessary because of oncological reasons. This algorithm has to be validated by a prospective analysis. PMID- 11435824 TI - A postoperative prognostic nomogram for renal cell carcinoma. AB - PURPOSE: Few published studies have combined prognostic factors to predict the likelihood of recurrence after surgery for renal cell carcinoma. We developed a nomogram for this purpose. MATERIALS AND METHODS: With Cox proportional hazards regression analysis, we modeled pathological data and disease followup for 601 patients with renal cell carcinoma who were treated with nephrectomy. Predictor variables were patient symptoms, including incidental, local or systemic, histology, including chromophobe, papillary or conventional, tumor size, and pathological stage. Treatment failure was recorded when there was either clinical evidence of disease recurrence or death from disease. Validation was performed with a statistical (bootstrapping) technique. RESULTS: Disease recurrence was noted in 66 of the 601 patients, and those in whom treatment was successful had a median and maximum followup of 40 and 123 months, respectively. The 5-year probability of freedom from failure for the patient cohort was 86% (95% confidence interval 82 to 89). With statistical validation, predictions by the nomogram appeared accurate and discriminating with an area under the receiver operating characteristic curve, that is a comparison of the predicted probability with the actual outcome of 0.74. CONCLUSIONS: A nomogram has been developed that can be used to predict the 5-year probability of treatment failure among patients with newly diagnosed renal cell carcinoma. The nomogram may be useful for patient counseling, clinical trial design and patient followup planning. PMID- 11435825 TI - Lack of retroperitoneal lymphadenopathy predicts survival of patients with metastatic renal cell carcinoma. AB - PURPOSE: Patients with metastatic renal cell carcinoma have a reported 5-year survival of 0% to 20%. The ability to predict which patients would benefit from nephrectomy and interleukin-2 (IL-2) therapy before any treatment is initiated would be useful for maximizing the advantage of therapy and improving the quality of life. MATERIALS AND METHODS: A retrospective analysis of the x-rays and charts of patients treated at the National Institutes of Health Surgery Branch between 1985 and 1996, who presented with metastatic renal cancer beyond the locoregional area and the primary tumor in place, was performed. Preoperative computerized tomography or magnetic resonance imaging, or radiological reports if no scans were available, were used to obtain an estimate of the volume of retroperitoneal lymphadenopathy. Operative notes were used to evaluate whether all lymphadenopathy was resected or disease left in situ, or if any extrarenal resection, including venacavotomy, was performed. Mean survival rate was calculated from the time of nephrectomy to the time of death or last clinical followup. If patients received IL-2 therapy, the response to treatment was recorded. Mean survival and response rate for IL-2 were compared among patients in 3 separate analyses. Patients without preoperatively detected lymphadenopathy were compared with those with at least 1 cm.3 retroperitoneal lymphadenopathy. Also, the patients who had detectable lymphadenopathy were divided into subgroups consisting of all resected, incompletely resected, unresectable and unknown if all disease was resected. Each subgroup was compared with patients without detectable preoperative lymphadenopathy. Patients with less than were compared to those with greater than 50 cm.3 retroperitoneal lymphadenopathy. Patients undergoing extrarenal resection at nephrectomy (complex surgery) due to direct invasion of the tumor into another intra-abdominal organ were compared with those undergoing radical nephrectomy alone, regardless of lymph node status. Statistical analysis was done with the Mantel-Cox test for comparison of survival on Kaplan-Meier curves and with Fisher's exact test for response rates for IL-2. RESULTS: A total of 154 patients with metastatic renal cell carcinoma underwent cytoreductive nephrectomy as preparation for IL-2 based regimens. There were 82 patients with metastatic renal cell carcinoma and no preoperative retroperitoneal lymphadenopathy who survived longer (median 14.7 months) than the 72 with lymphadenopathy (median 8.5, p = 0.0004). Patients with incompletely resected, unresectable or an unknown volume resected had decreased survival compared to those with no retroperitoneal lymphadenopathy. A multivariate analysis of survival was performed evaluating the known prognostic indicators, performance status and tumor burden, as represented by the number of organs involved with metastases, and the new prognostic factor, lymphadenopathy. Lymphadenopathy was more closely associated with survival than performance status, and appeared to be a new prognostic variable. Patients with and without retroperitoneal lymphadenopathy at initial presentation had similar rates for treatment with IL-2 (54% for both groups). Of the 82 patients with no lymphadenopathy 11 (13%) had long-term survival greater than 5 years. Of the 6 complete responses to IL-2, 5 occurred in this group. Only 1 other patient with incompletely resected retroperitoneal lymphadenopathy survived longer than 5 years. No significant difference in survival was seen between patients who did or did not undergo complex surgery. CONCLUSIONS: Patients who presented with metastatic renal cancer and retroperitoneal lymphadenopathy had a shorter survival than those with no detectable retroperitoneal lymphadenopathy. It is warranted to continue to perform complex extrarenal resection during nephrectomy since no significant difference in the response rate for IL-2 or mean survival compared with those of patients undergoing nephrectomy alone is currently detectable. PMID- 11435826 TI - Towards kidney cancer crystal ball. Better prognostication of patients with renal cell carcinoma. PMID- 11435827 TI - Screening and monitoring for bladder cancer: refining the use of NMP22. AB - PURPOSE: While detecting bladder cancer, bladder tumor markers demonstrate improved sensitivity compared with urinary cytology but the current limitation is the low specificity and positive predictive value, that is high false-positive rate. We examined the clinical categories of the false-positive results, established relative exclusion criteria, and recalculated the specificity and positive predictive value of this assay with these criteria. MATERIALS AND METHODS: A total of 608 patients considered at risk for bladder cancer presented to a urology clinic and submitted a single urine sample. Of the 608 patients 529 (87%) presented with de novo hematuria or chronic voiding symptoms without a diagnosis of bladder cancer. There were 79 (13.0%) patients being monitored with a known history of bladder cancer. Each urine sample was examined via cytology, urinalysis, culture and NMP22 protein assay. All patients underwent office cystoscopy, and transurethral resection and/or biopsy if a bladder tumor was suspected. RESULTS: Of the 608 patients 226 (37.2%) presented with microscopic hematuria, 143 (23.5%) with gross hematuria and 239 (39.3%) had chronic symptoms of urinary frequency or dysuria. There were 52 (8.6%) patients who had histologically confirmed bladder cancer. Of these 52 cancers NMP22 detected 46 (88.5%), whereas cytology identified only 16 (30.8%). When atypical cytology was considered positive, cytology detected 32 (61.5%) cases. In the 135 patients with increased NMP22 values the 46 identified tumors were accompanied by 89 false positive values yielding a specificity of 83.9% and a positive predictive value of 34.1%. These false-positive results were divided into 6 clinical categories. Exclusion of these categories improved the specificity and positive predictive value of NMP22 to 99.2% and 92.0%, respectively, yielding results similar to urinary cytology (99.8% and 94.1%). CONCLUSIONS: Awareness and exclusion of the categories of false-positive results can increase the specificity and positive predictive value of NMP22, enhancing the clinical use of this urinary tumor marker. PMID- 11435828 TI - Partial cystectomy during radical surgery for nonurological malignancy. AB - PURPOSE: It is not uncommon for a locally advanced, nonurological malignancy to invade the bladder. Partial cystectomy may be required to ensure complete tumor eradication. To our knowledge the true benefit of this procedure is unknown. MATERIALS AND METHODS: A total of 45 patients underwent partial cystectomy as part of radical surgery for a nonurological malignancy. We retrospectively reviewed these cases to determine which malignancies are prone to invade the bladder, the incidence of malignant invasion, the complication rate and the prognosis after wide en bloc resection. RESULTS: Colorectal adenocarcinoma accounted for the majority of cases. Tumor invaded only 11 bladder specimens (21.5%). Radical surgery was performed with curative intent in 30 patients, of whom 23 had negative surgical margins. At a mean followup of 30.7 months 16 of these 23 patients (69.6%) were free of disease or died of other disease processes. Disease progression and/or cancer related death occurred in 14 of the 15 patients (93.3%) who underwent surgery for palliation and in 16 of the 17 (94.1%) with positive margins at a mean of 21.7 months. Overall disease specific survival in those with malignant invasion was 27.3% compared to 41.2% when the bladder was fixed by a dense fibrous reaction only. No reported complications were related to partial cystectomy at followup. CONCLUSIONS: Advanced primary and recurrent nonurological malignancies often involve the bladder. Partial cystectomy may be necessary due to a dense fibrous reaction or direct tumor extension. While this distinction is made only after formal pathological results are reviewed, wide en bloc resection is necessary to ensure complete excision. When radical surgery is performed with curative intent and negative surgical margins are achieved, patients are likely to experience prolonged disease-free survival. PMID- 11435829 TI - Transrectal ultrasound guided biopsy of the prostate. Do enemas decrease clinically significant complications? AB - PURPOSE: Transrectal ultrasound guided biopsy of the prostate is the most common modality used to diagnose prostate cancer. Although many biopsy protocols have been described, in our opinion the role of enema before biopsy has not been definitively assessed in the literature. MATERIALS AND METHODS: A retrospective review of 448 transrectal ultrasound guided biopsies was performed. All biopsies were done with the same equipment, and all patients received identical antibiotic prophylaxis with ciprofloxacin. There were 38 patients excluded from the study secondary to alternate antibiotic prophylaxis. A total of 225 patients received enemas before biopsy, while 185 did not. Clinically significant complications necessitating office visit, secondary therapy and hospitalization were evaluated. RESULTS: Overall, clinically significant complications developed in 4.4% (10 of 225) of patients who had versus 3.2% (6 of 185) of those who did not have an enema (p = 0.614). There were 2 patients in each group who underwent transurethral prostatic resection or suprapubic prostatectomy for gross hematuria and/or urinary retention after biopsy. Of the patients who received enemas 2 were hospitalized for urinary retention and complicated urinary tract infection. One patient in the group without enema was hospitalized for gross hematuria and clot urinary retention. No patients who did not receive enema preparation were hospitalized for infectious complications. CONCLUSIONS: Transrectal ultrasound guided prostate biopsy accompanied by quinolone antibiotic prophylaxis remains a relatively safe procedure. Enema before biopsy provides no clinically significant outcome advantage, and potentially increases patient cost and discomfort. PMID- 11435830 TI - Prostate cancer diagnosis using a saturation needle biopsy technique after previous negative sextant biopsies. AB - PURPOSE: We hypothesized that markedly increasing the number of cores obtained during prostate needle biopsy may improve the cancer detection rate in men with persistent indications for repeat biopsy. MATERIALS AND METHODS: We performed saturation ultrasound guided transrectal prostate needle biopsy in 224 men under anesthesia in an outpatient surgical setting in whom previous negative biopsies had been performed in the office. The mean number of previous sextant biopsy sessions plus or minus standard deviation before saturation biopsy was 1.8 (range 1 to 7). A mean of 23 saturation biopsy cores (range 14 to 45) were distributed throughout the whole prostate, including the peripheral, medial and anterior regions. Indications for repeat biopsy were persistent elevated serum prostate specific antigen (PSA) in 108 cases, persistent elevated PSA and abnormal rectal examination in 27, persistent abnormal rectal examination in 4, high grade prostatic intraepithelial neoplasia in the previous biopsy in 64 and atypia in the previous biopsy in 21. RESULTS: Cancer was detected in 77 of 224 patients (34%). The number of previous negative sextant biopsies was not predictive of subsequent cancer detection by saturation biopsy. Median PSA was 8.7 ng./ml. and median PSA velocity was 0.63 ng./ml. yearly. Of the 77 patients in whom cancer was detected radical prostatectomy was performed in 52. Pathological stage was pT2 in 48 patients and pT3 in 4, while Gleason score was 4 to 5, 6 to 7 and 8 in 5, 46 and 1, respectively. At prostatectomy median cancer volume was 1.04 cc and 85.7% of removed tumors were clinically significant, assuming a 3-year doubling time. The overall complication rate for saturation needle biopsy was 12% and hematuria requiring hospital admission was the most common event. CONCLUSIONS: Saturation needle biopsy of the prostate is a useful diagnostic technique in men at risk for prostate cancer with previous negative office biopsies. This technique allows adequate sampling of the whole prostate gland and has a detection rate of 34% in this cohort of patients. PMID- 11435831 TI - Serial prostate specific antigen, free-to-total prostate specific antigen ratio and complexed prostate specific antigen for the diagnosis of prostate cancer. AB - PURPOSE: The free-to-total prostate specific antigen (PSA) ratio and complexed PSA have been introduced as adjuncts to total PSA for prostate cancer screening. Little data exist on the use of these tests for serial PSA screening. We compared serial total PSA, the free-to-total PSA ratio and calculated complexed PSA in men diagnosed with prostate cancer and matched controls in a population based study. MATERIALS AND METHODS: We identified 90 men diagnosed with prostate cancer between 1988 and 1996 with at least 3 serial serum samples obtained at 2-year intervals who were participants in the beta-Carotene and Retinol Efficacy Trial for the prevention of lung cancer. Samples were available up to 10 years before diagnosis. A total of 90 age matched men from the same cohort without prostate carcinoma were identified as controls. Free and total PSA was measured by the Abbott AxSYM system. RESULTS: Baseline demographics of cases and controls were similar. At baseline and diagnosis the men with prostate cancer had higher total and complexed PSA, and a lower free-to-total PSA ratio than controls. Mean followup was 5.2 years in cases and 5.5 in controls. The yearly change in PSA parameters in cases versus controls was 20.7% versus 3.5% for total, -3.4% versus 0.2% for free-to-total and 21.5% versus 3.4% for complexed PSA (p <0.0001). At diagnosis PSA alone was estimated to perform with more than 90% specificity in our model. CONCLUSIONS: In this population based study total PSA was superior to the free-to-total PSA ratio for predicting the development of prostate cancer. While serial changes in free-to-total PSA ratios with time were statistically significantly different in men diagnosed with prostate cancer and controls, the magnitude of these serial changes were slight enough to render them clinically insignificant. PMID- 11435832 TI - Prostate cancer detection in veterans with a history of Agent Orange exposure. AB - PURPOSE: Agent Orange, a chemical that was widely used in the Vietnam War as a defoliant, is widely accepted as a health hazard but its potential causative role in prostate cancer has been controversial. We evaluated the rate of prostate cancer in veterans referred for prostate biopsy who reported a history of Agent Orange exposure compared to the rate in veterans who denied such exposure. MATERIALS AND METHODS: A total of 400 consecutive veterans referred for prostate needle biopsy in a 30-month period completed a survey regarding Agent Orange exposure. Of these 400 patients 32 (8%) reported previous exposure to Agent Orange. From the remaining 368 patients who denied Agent Orange exposure 3 consecutive age matched controls were selected per each patient reporting exposure for a total of 96 age matched controls. Prostate specific antigen, prostate cancer, cancer grade and length of cancer in the biopsy cores were compared in Agent Orange exposed patients and unexposed controls. To determine whether the patient population referred for biopsy was skewed by proportionally more exposed and referred than unexposed patients those referred for biopsy were compared to the overall adult male veteran population followed at the outpatient clinics at our facility. RESULTS: Of the 32 Agent Orange exposed patients 13 (41%) had prostate cancer, while 33 of the 96 controls (34.4%) had cancer. There was no correlation of Agent Orange exposure with cancer (r = 0.06). There was also no statistically significant difference in the 2 groups in regard to PSA (p = 0.90), cancer (p = 0.15), proportion of well differentiated cancers (p = 0.41) or length of cancer in the biopsy cores (p = 0.34). Compared with the total adult male veteran population followed on an outpatient basis at our facility an average of 1.07% of those with a history of Agent Orange exposure were referred for prostate biopsy yearly versus 1.33% of unexposed patients. CONCLUSIONS: Agent Orange may have a role in the causation of some types of cancer but we identified no significant relationship of prostate cancer with Agent Orange exposure in patients referred for prostate biopsy. PMID- 11435833 TI - Relationship between systematic biopsies and histological features of 222 radical prostatectomy specimens: lack of prediction of tumor significance for men with nonpalpable prostate cancer. AB - PURPOSE: Because of the recent increase in nonpalpable prostate cancer (clinical stage T1c) in men, preoperative needle biopsy findings have had an important role for treatment decisions. We examine the correlation among histopathological features of 6 systematic biopsies and radical prostatectomy specimens in which 1 investigator reviewed all histological sections. MATERIALS AND METHODS: We studied a total of 450 men with clinical stage T1c prostate cancer from whom needle biopsies were matched with radical prostatectomy specimens, and selected 222 patient biopsies that were obtained from 6 or more separate regions of the prostate. The pretreatment parameters of serum prostate specific antigen (PSA), PSA density, number of positive needle biopsies, distribution of positive cores, linear cancer length, and percent Gleason grade 4/5 on the biopsy were determined and compared with histopathological features of prostate cancer in the radical prostatectomy specimens. All biopsies and radical prostatectomies were evaluated morphologically at the department of urology. RESULTS: Of the 222 men the largest cancer was clinically insignificant in 23 (10%), as measured by a cancer volume of less than 0.5 cc. Cancer volume in the prostatectomy specimen was significantly related to all parameters in the biopsy, with the surprising exception of cancer distribution in the positive biopsies. However, all of these correlations with cancer volume were weak, with Pearson's correlation squared (R(2)) multiplied by 100 less than 10%. Unfortunately, tumor grade on the biopsy agreed with the prostatectomy specimen in only 81 of 222 (36%) cases. Grade assessment with needle biopsy underestimated the tumor grade in 102 (46%) cases and overestimated it in 39 (18%). No single parameter in the biopsy was a predictor of tumor significance, as measured by a cancer volume of greater than 0.5 cc. However, the best model to predict a tumor less than 0.5 cc in volume was the combination of a single positive core with cancer length less than 3 mm. that contained no Gleason grade 4/5. The use of PSA or PSA density in combination with needle biopsy findings did not enhance prediction of tumor significance. CONCLUSIONS: These results indicate a weak and disappointing correlation among all pathological features of 6 systematic biopsies and radical prostatectomy specimens. The combination of 1 positive core with cancer length less than 3 mm. that contains no Gleason grade 4/5 is probably the best predictor of prostate cancer less than 0.5 cc in men with nonpalpable tumors, a cancer volume that occurred in only 10% of the 222 (23) men. PMID- 11435834 TI - Is anastomotic biopsy necessary before radiotherapy after radical prostatectomy? AB - PURPOSE: External beam radiotherapy may be given after radical prostatectomy as adjuvant (immediate) or therapeutic (delayed) treatment, the latter in response to evidence of disease recurrence. In patients receiving delayed radiotherapy the necessity of a positive anastomotic biopsy before treatment remains unclear. We determined whether a positive anastomotic biopsy predicted the response to radiation in this setting. MATERIALS AND METHODS: We reviewed the records of 67 patients who received radiotherapy for biochemical or biopsy proved recurrent prostate cancer after radical prostatectomy. Patients underwent surgery at our institution or its affiliated hospitals, or were referred to our institution for radiotherapy. All patients had a negative metastatic evaluation before receiving radiotherapy. Biochemical failure after radiotherapy was defined as serum prostate specific antigen (PSA) 0.2 ng./dl. or greater on 2 or more consecutive occasions. Biochemical recurrence-free survival was calculated using the Kaplan Meier method. Independent predictors of PSA failure after radiotherapy were identified using the multivariate Cox proportional hazards model. RESULTS: Of the 67 patients evaluated 33 and 34 received radiotherapy for biochemical failure and biopsy proved local recurrence, respectively. The 3-year recurrence-free survival rate was 49% in patients treated for biochemical failure and 39% in those with biopsy proved local recurrence. There was no significant difference in PSA-free survival in these 2 groups. Only pre-radiotherapy PSA 1 ng./dl. or greater (p = 0.02) and seminal vesicle invasion (p = 0.02) were significant independent predictors of biochemical failure. CONCLUSIONS: A positive anastomotic biopsy did not predict an improved outcome after radiotherapy following radical prostatectomy. Anastomotic biopsy was associated with a longer time to salvage radiotherapy. However, this delay did not translate into worse disease-free outcomes in patients who underwent anastomotic biopsy. High pre-radiotherapy PSA greater than 1 ng./ml. was the most significant predictor of biochemical failure after therapeutic radiotherapy. Decisions regarding local radiation therapy after radical prostatectomy may be made without documenting recurrent local disease. PMID- 11435835 TI - 20-year outcome of patients with T1-3N0 surgically staged prostate cancer treated with external beam radiation therapy. AB - PURPOSE: Patients with surgically staged localized prostate cancer treated with external beam radiation therapy were retrospectively analyzed for 15 and 20-year overall and cause specific survival. The need for additional therapy was also evaluated. MATERIALS AND METHODS: We analyzed 145 patients who received external beam radiotherapy after negative staging pelvic lymphadenectomy. Followup data were available for 129 patients. Overall and cause specific survival was calculated with the Kaplan-Meier method. RESULTS: Median followup was 14.9 years. Actuarial overall survival at 15 and 20 years was 45.9% and 24.6%, respectively. Cause specific survival at 15 and 20 years was 64.5% and 37.7% for having all patients dying of unknown causes censored, and 54.4% and 30.1% for those dying of unknown causes categorized as having prostate cancer, respectively. Of the patients who survived 47% were on hormonal therapy. CONCLUSIONS: Longer followup after external beam radiation therapy continues to demonstrate a decrease in cause specific survival. Many patients ultimately require hormonal therapy. PMID- 11435836 TI - Gunshot wounds to the ureter: a 40-year experience at Grady Memorial Hospital. AB - PURPOSE: Because of the morbidity associated with missed ureteral injuries, a high index of suspicion must be present to diagnose this type of injury. We reviewed our 40 years of trauma experience at Grady Memorial Hospital. MATERIALS AND METHODS: We retrospectively reviewed the records of 118 patients with ureteral injuries secondary to a gunshot wound to the ureter from 1960 to 1999. All cases were reviewed for the mechanism of injury, location, initial urinalysis, imaging modalities, associated injuries, operative procedures and complications. RESULTS: Our population consisted of patients 14 to 71 years old, of whom 66 had right ureteral, 51 had left ureteral and 1 had bilateral injury. Urinalysis revealed no evidence of blood in the urine in 15% of the patients in whom the test was performed. Excretory urography had a false-negative rate of 33%. The injury was located at the proximal, mid and distal ureter in 43, 38 and 37 cases, respectively. Multiple surgical approaches were used depending on the location and severity of the defect. Only 1 patient had an isolated ureteral injury, while the remainder had associated injuries. Complications were present in 24 cases. CONCLUSIONS: To our knowledge this series of ureteral injuries is the largest reported to date. Because preoperative urinalysis and imaging studies are unreliable for ruling out injury, a high index of suspicion must be present. Furthermore, a predefined trauma protocol, as defined in our algorithm, decreases the number of missed ureteral injuries that may potentially complicate the outcome of an already critical case. PMID- 11435837 TI - Genetic material is present in cadaveric dermis and cadaveric fascia lata. AB - PURPOSE: We determined whether genetic material is present in the commercially processed cadaveric allografts used in sling surgery. MATERIALS AND METHODS: We evaluated 16 samples from 2 commercial sources of human allograft, including 8 each of freeze-dried gamma irradiated cadaveric fascia lata and acellular cadaveric dermis. Fresh human rectus fascia and sterile saline served as positive and negative controls, respectively. All samples underwent a standard proteinase K/sodium dodecyl sulfate/phenol extraction technique to isolate DNA. Polymerase chain reaction was done to amplify the retrieved DNA material, spectrophotometry to quantify DNA concentration and agarose gel electrophoresis to determine the size of DNA fragments. RESULTS: Of the 16 samples tested from 2 commercial sources of human allograft fascia 14 (87.5%) contained DNA. Mean DNA concentration plus or minus standard error was 258.3 +/- 80.1 and 272.8 +/- 168.8 microg./gm. tissue for cadaveric fascia lata and cadaveric dermis, respectively. Polymerase chain reaction amplified DNA segments of 2,000 bp from 1 of each of the 8 samples of cadaveric fascia lata and cadaveric dermis. CONCLUSIONS: Freeze dried gamma irradiated cadaveric fascia lata and acellular cadaveric dermis contained intact DNA. PMID- 11435838 TI - Interexaminer reliability of transrectal ultrasound for estimating prostate volume. AB - PURPOSE: We investigated the interexaminer reliability of transrectal ultrasound measurement of total prostate and transition zone volume among 3 examiners with various levels of experience. MATERIALS AND METHODS: A total of 121 patients 39 to 82 years old (average plus or minus standard deviation 60.7 +/- 10.3) from a single urology clinic volunteered to participate. Patients with prostate cancer, previous prostate surgery or recent invasive prostatic examination were excluded from study. Each individual was examined independently by each of 3 examiners with various levels of experience, including an attending urologist, a PGY-2 resident in the second year of general surgery before urology training and a PGY 4 resident in the second year of urology training. Transrectal ultrasound was performed in each case by each examiner in pre-specified random order. RESULTS: Mean total prostate and transition zone volume was 35.9 +/- 27.2 and 15.6 +/- 18.8 ml., respectively. Interexaminer agreement or reliability of the ultrasound measurements was high for total prostate and transition zone volume (intraclass correlation 0.96, 95% confidence interval [CI] 0.95 to 0.97 and 0.93, 95% CI 0.90 to 0.95, respectively). For individual prostatic dimensions reliability estimates were 0.78 to 0.86, while for transition zone dimensions reliability was 0.85 to 0.90. Total prostate volume reliability was higher for prostate volume greater than 40 ml. versus smaller prostates (intraclass correlation 0.95, 95% CI 0.90 to 0.97 versus 0.77, 95% CI 0.67 to 0.84). Mean differences in transrectal ultrasound measurements by different examiners were highest for the resident with least experience. CONCLUSIONS: The reliability of transrectal ultrasound measured total prostate and transition zone volume is high for examiners with different levels of experience at this institution. Reliability in patients without prostate cancer appears to be better for larger volume prostates and for examiners with more experience. PMID- 11435839 TI - Intermittent catheterization with a prelubricated catheter in spinal cord injured patients: a prospective randomized crossover study. AB - PURPOSE: We compared the safety and patient acceptance of a conventional Nelaton and a prelubricated nonhydrophilic catheter in 18 spinal cord injured patients on intermittent catheterization. MATERIALS AND METHODS: In a prospective crossover study each catheter was used for 7 weeks and the initial course was randomized. Urinalysis and urine culture were performed at 2, 4 and 7 weeks. Urethral trauma was evaluated by urethral cell count on the surface of each catheter used on the last day of each study period. Patient satisfaction was assessed at the end of the study by a questionnaire using multiple visual analog scales. RESULTS: Urinary tract infection was identified in 12 and 4 patients on a Nelaton and a prelubricated nonhydrophilic catheter (p = 0.03), while asymptomatic bacteruria was identified in 18 and 8 (p = 0.0244), respectively. The mean urethral cell count plus or minus standard deviation on the catheter surface was 6.7 +/- 2.8 x 10(4) and 15.1 +/- 8.9 x 10(4) for the prelubricated nonhydrophilic and the Neelaton catheter, respectively (p = 0.01). The prelubricated nonhydrophilic catheter resulted in a better mean satisfaction score than the Nelaton catheter (2.33 +/- 1.06 versus 4.72 +/- 2.13, p = 0.022). Urethral bleeding was reported in 2 patients during the study period while using the Nelaton catheter. CONCLUSIONS: The prelubricated nonhydrophilic catheter is a safe, effective and comfortable option in spinal cord injured patients on intermittent self catheterization. PMID- 11435840 TI - Use of the neodymium: YAG laser for interstitial cystitis: a prospective study. AB - PURPOSE: Interstitial cystitis is a disorder of the bladder characterized by urgency and frequency of urination, and pelvic pain. The classic type of interstitial cystitis is characterized by Hunner's ulcers, which are focal regions of severe bladder inflammation. Patients with Hunner's ulcers tend to have more severe symptoms and are often refractory to medical management. We present a prospective series of patients who underwent ablative therapy of Hunner's ulcers using a neodymium (Nd):YAG laser. MATERIALS AND METHODS: A total of 24 patients with interstitial cystitis underwent ablative therapy for Hunner's ulcers. Medical therapy had failed in all cases. Using regional or general anesthesia the Nd:YAG laser under cystoscopic control was used to ablate the ulcers. The power setting was 15 W. with a firing duration of between 1 and 3 seconds. The procedure was performed on an outpatient basis. Symptoms were noted preoperatively and postoperatively. RESULTS: All patients had symptom improvement within 2 to 3 days. The mean pain scores decreased from 9.1 to 1.2 (p <0.003), the mean urgency score decreased from 8.2 to 1.9 (p <0.003), the mean voiding interval increased from every 30 minutes to every 102 (p <0.0001) and nocturia decreased from a mean of 7.9 voids per night to 2.9 (p <0.0001). There were no complications. Mean followup was 23 months. However, relapse in 11 patients required 1 to 4 additional treatments. The re-treatment response was similar to the initial treatment. CONCLUSIONS: Nd:YAG laser ablation of Hunner's ulcers is an excellent, minimally invasive method of treating interstitial cystitis. While it is not a cure, it offers patients an opportunity to have decreased symptoms for an extended period and it may be repeated as necessary. PMID- 11435841 TI - Percutaneous aspiration and sclerotherapy for treatment of spermatoceles. AB - PURPOSE: Spermatoceles are commonly treated with open surgical ablation. With the increasing shift towards minimally invasive surgery and heightened awareness, both patient and surgeon accept and prefer the less intrusive approach. Treatment of spermatoceles translates into the use of aspiration and sclerotherapy rather than open excision. We report on our prospective study of percutaneous aspiration and sclerotherapy with sodium tetradecyl sulfate for treatment of spermatoceles to determine safety, efficacy and cost-effectiveness. MATERIALS AND METHODS: Patients with symptomatic cystic scrotal lesions were prospectively enrolled in an aspiration and sclerotherapy protocol between October 1998 and June 2000. All patients underwent aspiration followed by sclerotherapy with a sodium tetradecyl sulfate solution. Followup included clinical assessment and scrotal ultrasonography at 12 weeks and on a needed basis thereafter. RESULTS: A total of 28 patients with 34 spermatoceles were enrolled in the protocol. Mean patient age was 61.0 years and mean spermatocele volume was 89.3 ml. Complete followup was available for 31 of 34 (91%) spermatoceles, and the mean duration of followup was 24.6 weeks. Results were cure in 29% success in 36%, partial success in 19% and failure in 16% of cases. Patient satisfaction was 85% and there were no complications. The total cost of each procedure was only $104 (Canadian), and none took more than 10 minutes. CONCLUSIONS: Percutaneous aspiration and sclerotherapy with sodium tetradecyl sulfate is a minimally invasive, simple, safe, inexpensive and reasonably efficacious treatment option for spermatoceles. Initial failures can be salvaged with second treatment. PMID- 11435842 TI - A short-term, multicenter, randomized double-blind dose titration study of the efficacy and anticholinergic side effects of transdermal compared to immediate release oral oxybutynin treatment of patients with urge urinary incontinence. AB - PURPOSE: We compared the short-term efficacy, safety and tolerability of transdermal versus oral oxybutynin in adults with urge urinary incontinence. MATERIALS AND METHODS: Volunteers with detrusor instability currently responding to oral immediate release oxybutynin were enrolled in our study. Those patients presenting with recurrence of incontinent symptoms after a 2-week washout underwent confirmatory cystometrogram with subsequent randomization to transdermal or oral treatment. Matching active and placebo medications included matrix patches applied twice weekly and capsules taken 2 or 3 times daily. Dose titration was based on anticholinergic symptoms. Outcome measures included comparison of baseline to 6 week changes in incontinence episodes on a 3 day urinary diary, a visual analog scale for efficacy and anticholinergic symptoms reported on a questionnaire. Safety monitoring included adverse events and skin tolerability of the transdermal system. RESULTS: A total of 76 patients were enrolled and 74 completed at least 4 weeks of treatment. Mean age in the transdermal and oral groups was 64 and 63 years, and 87% and 97% were female, respectively. Daily incontinent episodes decreased in the transdermal and oral groups (7.3 to 2.4 [66%] and 7.4 to 2.6 [72%], respectively, p = 0.39). The visual analog scale reduction in urinary leakage improved from washout in both groups (p <0.0001) with no difference between them (p = 0.9). Dry mouth occurred in significantly fewer patients in the transdermal (38%) compared with those in the oral group (94%, p <0.001). Of the patients in the transdermal group 67% noticed a reduction in dry mouth severity compared with previous oral treatment, and 90% had none or mild skin erythema. CONCLUSIONS: Transdermal delivery of oxybutynin resulted in comparable efficacy and a significantly improved anticholinergic side effect profile compared with oral administration in adults with urge urinary incontinence. PMID- 11435843 TI - A crossover randomized trial of transcutaneous electrical nerve stimulation and oxybutynin in patients with detrusor instability. AB - PURPOSE: Management of idiopathic detrusor instability is difficult in most patients mainly due to the lack of a complete understanding of the pathophysiology. Oxybutynin and transcutaneous electrical nerve stimulation have been used but to our knowledge no direct comparisons have been made. MATERIALS AND METHODS: Patients with frequency, urgency, urge incontinence and proved detrusor instability were studied with urodynamics, quality of life instruments, and frequency and volume charts. Patients were randomized to transcutaneous electrical nerve stimulation or oxybutynin. After 6 weeks of treatment, they were reassessed and after a washout of 2 weeks, they were started on the second arm of treatment and reassessed 6 weeks later. RESULTS: A total of 13 male and 30 female patients were studied. Functional capacity had increased and number of voids daily had decreased significantly compared with before treatment in both arms (p <0.005). There were significant improvements in symptom specific quality of life measures but no changes were found on the global Short Form 36 (SF-36) quality of life questionnaire. The volume to first desire to void and first unstable contraction had increased significantly with oxybutynin but not with transcutaneous electrical nerve stimulation. Of 23 patients 7 were stabilized with treatment, including 2 with oxybutynin only, 2 with either nerve stimulation or oxybutynin and the remaining 3 with only nerve stimulation. Total bladder capacity did not change significantly with either treatment but patients noticed side effects more commonly with oxybutynin. CONCLUSIONS: Both treatments clearly improved subjective parameters. However, only oxybutynin showed significant improvements in objective urodynamic parameters. Transcutaneous electrical nerve stimulation can be used in patients who cannot take oxybutynin. Further studies are needed to show the long-term efficacy and cost analyses of nerve stimulation. PMID- 11435844 TI - Improving conservative treatment of overactive bladder and urge incontinence--1 small step at a time. PMID- 11435845 TI - Anatomical and functional results of pubovaginal sling procedure using polypropylene mesh for the treatment of stress urinary incontinence. AB - PURPOSE: The anatomical and functional results of pubovaginal sling procedure using polypropylene mesh were investigated. MATERIALS AND METHODS: A total of 50 women with stress urinary incontinence, including 26 with type 2, 7 with mixed type 2/3 and 17 with type 3 disease, underwent the pubovaginal sling procedure using polypropylene mesh. A self-fashioned 2 cm. polypropylene mesh sling was used in all cases. The slings were fixed without tension at each side of the bladder neck. The sling was further suspended to elevate the bladder neck in patients with type 2 disease and those with a hypermobile bladder base, while it was placed without further tension in those with type 3 disease and those with a nonmobile bladder base. After the operation video urodynamics and transrectal sonography were done to evaluate functional and anatomical results. RESULTS: At a median followup of 24 months 42 women were cured, 6 had improvement and treatment had failed in 2. Although the maximum flow rate increased, no changes were noted in detrusor pressure, post-void residual urine or cystometric capacity at month 3. Transrectal sonography revealed that mean sling width plus or minus standard deviation had decreased to 13.5 +/- 3.3 mm. at 3 months. The endopelvic fascia was thicker in women with type 2 than in those with types 2/3 or 3 disease (mean 5.3 +/- 1.5 versus 3.5 +/- 1.3 mm., p = 0.001). The position of bladder neck on video urodynamics was elevated a mean of 2.01 +/- 2.01 and 3.07 +/- 2.07 cm. compared to baseline elevation during resting and straining, respectively (p = 0). No hypermobility was noted while the patients coughed and no kinking of the urethra was observed while they performed the Valsalva maneuver. Detrusor instability resolved in 6 of 8 women (75%) and new onset detrusor instability was noted in 7 (14%). In 2 women with persistent stress urinary incontinence a repeat sling procedure was necessary to treat type 3 disease and cure incontinence. The overall success rate was 96%. One patient with sling erosion into the vaginal wall was treated with simple revision. CONCLUSIONS: Fixation of a polypropylene mesh sling at the bladder neck and adequate suspension may effectively achieve a hammock effect without creating bladder outlet obstruction in patients with types 2 and 2/3 stress urinary incontinence. The anatomical and functional results of this study show that a well suspended bladder neck did not create bladder outlet obstruction after a pubovaginal sling procedure using polypropylene mesh. PMID- 11435846 TI - The intravesical potassium sensitivity test and urodynamics: implications in a large cohort of patients with lower urinary tract symptoms. AB - PURPOSE: Bladder outlet obstruction secondary to benign prostatic hyperplasia (BPH) is the most common diagnosis in older men with lower urinary tract symptoms. However, these symptoms also can occur with interstitial cystitis. We determine whether the potassium sensitivity test is useful for distinguishing BPH from possible intersitial cystitis in patients with lower urinary tract symptoms. We also test the hypothesis that patients with these symptoms who have a positive test will have urodynamic findings consistent with the diagnosis of interstitial cystitis. MATERIALS AND METHODS: The potassium sensitivity test was performed in 526 (95%) males and 25 (5%) females with lower urinary tract symptoms undergoing urodynamic testing. Urodynamic parameters in the positive and negative potassium sensitivity test groups were compared. % RESULTS: Of the patients 16% (89 of 551) had a positive potassium sensitivity test. Compared with patients who had a negative test, those who had a positive test were younger (61 versus 64 years, p = 0.03), had urgency at significantly lower volumes (108 versus 182 cc, p <0.0001), lower bladder capacity (343 versus 436 cc, p <0.0001) and lower post void residual (49 versus 95 cc, respectively, p <0.001). Urodynamic parameters in the 24% (6 of 25) of women who had a positive potassium sensitivity test were similar to those in men who also had a positive test. CONCLUSIONS: Urodynamic findings in patients with lower urinary tract symptoms who have a positive potassium sensitivity test are significantly different from those in patients who have a negative test, and are similar to those findings characteristic of interstitial cystitis. Interstitial cystitis should be considered in patients with lower urinary tract symptoms who have a positive test. The potassium sensitivity and urodynamic tests may be a useful combination for screening men with lower urinary tract symptoms to identify those symptoms that may be due to interstitial cystitis versus BPH. PMID- 11435847 TI - Smoking increases the risk of large scale fluid absorption during transurethral prostatic resection. AB - PURPOSE: We studied the association between smoking habits and absorption during transurethral prostatic resection in healthy patients, and in those with medical disease and prostate cancer. MATERIALS AND METHODS: Smoking habits and the presence of medical disease and prostate cancer in 1,034 patients who underwent transurethral prostatic resection were compared with fluid absorption levels. These levels were measured by the ethanol method and surgical blood loss, as determined by a photometer. RESULTS: The incidence of fluid absorption greater than 1,000 ml. was 15.7% in the present smokers, 15.5% in the past and present smokers, and 8.8% in those who had never smoked. Compared to nonsmokers, the relative risk of 1,000 to 2,000 ml. fluid absorbed in past and present smokers was 1.8 (95% confidence interval 1.1 to 2.9). The corresponding relative risk of absorption greater than 2,000 ml. was 2.1 (95% confidence interval 1.1 to 4.3), with the greatest excess risk in present smokers, which was 2.8 (1.3 to 5.9). These differences between smokers and nonsmokers were even greater in the healthiest patients, while being relatively smaller in those with medical disease or prostate cancer. Fluid absorption correlated with the amount of blood lost but the demonstrated association between smoking and fluid absorption (p <0.024) could not be explained by an effect on blood loss. CONCLUSIONS: Past and present smoking increases the risk of large scale fluid absorption during transurethral prostatic resection. A possible reason is that smoking alters vascular growth in the gland. PMID- 11435848 TI - Transurethral prostatic resection or laser therapy for men with acute urinary retention: the ClasP randomized trial. AB - PURPOSE: Transurethral resection of the prostate is the standard operation for acute urinary retention, although laser prostatectomy is reportedly effective and safe. The ClasP (conservative management, laser, transurethral resection of the prostate) study compared transurethral prostatic resection and noncontact neodymium (Nd):YAG visual laser assisted prostatectomy for treatment of acute urinary retention. MATERIALS AND METHODS: This study was a multicenter randomized controlled trial, analyses were by intention to treat and followup was at 7.5 months after randomization. Primary outcomes were treatment failure, and included International Prostate Symptom Score, International Prostate Symptom Score quality of life score, residual urine and flow rate. Secondary outcomes included complications, and duration of catheterization and hospitalization. RESULTS: A total of 148 men were randomized to transurethral prostatic resection (74) and laser (74). There were fewer treatment failures after prostatic resection (p = 0.008) and fewer men after resection required secondary surgery for poor results (1 versus 7, p = 0.029). Maximum flow rates after transurethral prostatic resection were better than after laser (mean difference 4.4 ml. per second). Comparison of symptom and quality of life scores demonstrated that any clinically significant advantage for laser could be ruled out. Patients stayed a mean of 2 extra days in the hospital after resection. The duration of catheterization was greater after laser but significantly fewer major treatment complications were found with laser therapy. CONCLUSIONS: Transurethral prostatic resection was more effective, resulted in fewer failures than laser treatment and remains the procedure of choice for men with acute urinary retention. PMID- 11435849 TI - A prospective study of the safety and efficacy of suprapubic transvesical prostatectomy in patients with benign prostatic hyperplasia. AB - PURPOSE: We investigate the safety and efficacy of suprapubic transvesical prostatectomy, and the change in bladder wall thickness after surgery. MATERIALS AND METHODS: We conducted a prospective 1 center study of 32 consecutive patients who underwent transvesical prostatectomy from December 1996 to March 1997 for benign prostatic hyperplasia with large prostate volume, who were followed for 1 year. Pressure flow study and transrectal sonography were performed at baseline and repeated at 6 months. Bladder wall thickness was measured at baseline and regular intervals postoperatively. A morbidity questionnaire was completed during the first 6 weeks after surgery. RESULTS: An average of 63 gm. prostate adenoma were enucleated at surgery. An indwelling catheter was required for an average plus or minus standard deviation of 5.4 +/- 2.6 days after treatment. The International Prostate Symptom Score decreased from 19.9 +/- 4.4 to 1.5 +/- 2.7 and the quality of life score decreased from 4.9 +/- 1.0 to 0.2 +/- 0.4 at year 1, respectively. Maximum flow rate improved from 9.1 +/- 5.3 to 29.0 +/- 8.9 ml. per second. Residual urine decreased from 128 +/- 113 to 8 +/- 18 ml. Before surgery 30 patients had obstruction and 2 were in the equivocal zone of the International Continence Society nomogram. At 6 months after prostatectomy 30 patients did not have obstruction, and 2 who were subsequently operated on for bladder neck sclerosis were equivocal and had obstruction, respectively. No patient had significant postoperative bleeding and no heterologous blood transfusions were required. There were 4 men who had urinary tract infection and 1 who had wound infection. A slight decrease in erectile function was observed 6 weeks postoperatively, and no change in patient libido and quality of sex life was reported. The total complication rate was 31.3%. The bladder was unstable in 7 men before and 3 after surgery. A significant decrease in bladder wall thickness was observed from 5.2 +/- 0.7 at baseline to 2.9 +/- 0.9 mm. at year 1 postoperatively. CONCLUSIONS: Our study confirms the excellent clinical outcome of transvesical prostatectomy, and rapid improvement of most subjective and objective parameters during the 4 weeks after surgery. Bladder hypertrophy appears to be significantly reduced after prostate surgery. The urodynamic results in patients who underwent open surgery probably represent the maximum obtainable relief of obstruction and should be considered the reference standard to which all other treatments, including transurethral resection, should aspire. PMID- 11435850 TI - Surgical management of benign prostatic hyperplasia in 2001--a pause for thought. PMID- 11435851 TI - Significant medical pathology discovered during a male infertility evaluation. AB - PURPOSE: Because a pregnancy can be achieved without a male infertility evaluation, some have questioned its usefulness. However, by bypassing a urological evaluation the man might not learn the cause of infertility and not be offered specific corrective therapy. In addition, men with subfertility may have a serious underlying medical or genetic problem that could also be overlooked. We determine the incidence of significant medical pathology discovered during a male infertility evaluation at 2 academic infertility practices. MATERIALS AND METHODS: All men examined for either primary or secondary infertility were included in our study, while men seen for vasectomy reversal were not. All patients underwent evaluation, consisting of a complete history, physical examination, semen analysis, hormone testing, urinalysis and genetic testing when appropriate. RESULTS: Significant medical pathology was discovered in 33 of 536 (6%) patients. A total of 27 patients had genetic abnormalities, including cystic fibrosis mutations in 24 and karyotypic abnormalities in 3. Of the remaining 6 patients 1 had testis cancer, 1 prostate cancer, 3 diabetes mellitus and 1 hypothyroidism. CONCLUSIONS: Significant medical pathology can be detected by a male infertility evaluation. In addition to identifying the cause of infertility, the evaluation may uncover conditions that threaten the health of the male partner or any potential offspring. PMID- 11435852 TI - A new method for the evaluation of erectile dysfunction: sildenafil plus Doppler ultrasonography. AB - PURPOSE: Of the various methods of hemodynamic studies performed to evaluate erectile dysfunction penile color Doppler ultrasound is currently considered the best. However, intracavernous injection is invasive and has adverse effects, such as prolonged erection. We evaluated whether sildenafil may be used as a substitute for intracavernous agents when assessing impotence on color Doppler ultrasound. MATERIALS AND METHODS: A total of 42 patients with erectile dysfunction underwent color Doppler ultrasound before and after intracavernous injection of 60 mg. papaverine with genital and audiovisual sexual stimulation. Peak flow and end diastolic velocity were measured in the recorded waveforms obtained 0, 1, 5, 10 and 20 minutes after injection. The patients also underwent color Doppler ultrasound after a 50 mg. oral dose of sildenafil with genital and audiovisual sexual stimulation not before 3 days after the papaverine study. The same parameters were measured at 30, 45, 60, 75 and 90 minutes, and compared with the values obtained after papaverine injection. RESULTS: Mean peak flow velocity significantly increased after oral sildenafil starting at 30 minutes and achieving a maximum value at 60 minutes. There were no significant differences in the 2 methods in mean peak velocity 1, 5, 10 and 20 minutes after papaverine injection, and 30, 45, 60, 75 and 90 minutes after oral sildenafil administration. Penile color Doppler ultrasound with intracavernous papaverine injection is accepted as the gold standard but color Doppler ultrasound with sildenafil has 90% sensitivity and 100% selectivity for demonstrating arterial insufficiency. Due to prolonged erection 5 patients (11.9%) in the papaverine group required pharmacological detumescence by intracavernous injection. No adverse effects of sildenafil were observed. CONCLUSIONS: Sildenafil administration achieved increased peak flow velocity comparable to that after intracavernous papaverine injection. With no prolonged erection sildenafil emerges as a safer alternative compared to more invasive intracavernous injection. PMID- 11435853 TI - A comparison of dermal and cadaveric pericardial grafts in the modified Horton Devine procedure for Peyronie's disease. AB - PURPOSE: Penile curvature due to Peyronie's disease is often treated with plaque incision or excision and grafting. The advantages of various graft materials have long been debated. We studied the outcomes of dermal and cadaveric pericardial grafts after plaque incision or excision for Peyronie's disease. MATERIALS AND METHODS: We retrospectively evaluated all modified Horton-Devine procedures performed between February 1998 and April 2000. Dermal graft harvested from the lower abdomen or commercially available cadaveric pericardium was used to cover the corporeal defect. A followup telephone questionnaire was administered to determine patient satisfaction and function. RESULTS: We reviewed the records of 27 men, including 18 and 9 who received a dermal and pericardial graft, respectively. We successfully contacted 24 patients, including 15 with a dermal and 9 with a pericardial graft, to evaluate satisfaction. Average postoperative followup was 10 months. Of the 24 patients, 22 reported successful coitus and 15 (63%) needed assistance to achieve erection rigid enough for satisfactory coitus. Eight patients reported recurrent curvature, which was minimal and did not hamper sexual function. Overall 11 of the 15 patients with a dermal and 8 of the 9 with a pericardial graft would undergo the surgery again or recommend it to others. CONCLUSIONS: The results of the dermal and cadaveric pericardial grafts were comparable. The minimal preoperative preparation, decreased patient morbidity and pliability of cadaveric pericardium make it an attractive suitable graft substitute in the modified Horton-Devine procedure. Longer followup and larger patient series are needed to confirm the advantages of cadaveric pericardium. PMID- 11435854 TI - Indications for admission to the surgical intensive care unit after radical cystectomy and urinary diversion. AB - PURPOSE: We analyzed the practice of mandatory surgical intensive care unit admission after radical cystectomy, and defined objective criteria to predict active treatment requirements and surgical intensive care unit stay. MATERIALS AND METHODS: We retrospectively reviewed the records of 115 consecutive patients admitted to the surgical intensive care unit after radical cystectomy and urinary diversion during the 36-month study period of January 1996 to December 1998. An Acute Physiology and Chronic Health Evaluation II score was calculated from postoperative patient parameters at admission to the unit. Active treatment mandating admission was defined as postoperative invasive cardiopulmonary monitoring, administration of vasopressors or inotropic medications, monitoring or treatment for life threatening complications, or mechanical ventilation for longer than 12 hours. We analyzed the correlation of outcome variables with the requirements for active treatment and surgical intensive care unit stay, and developed a stratification model of low versus high risk. Low risk was defined as a calculated likelihood of less than 10% for requiring active treatment postoperatively. RESULTS: Mean stay in the surgical intensive care unit plus or minus standard error was 34.4 +/- 3.1 hours. No active treatment was required in 63.5% of patients during the stay. The evaluation score, intraoperative complications and number of intraoperative transfusions were the strongest predictors of required postoperative active treatment. By combining these variables we developed a clinically applicable algorithm to stratify patients into a low and a high risk category. In patients at low and high risk the active treatment rate was 5.9% and 42.8% (p = 0.001), and the mean stay was 24.6 +/- 2.2 and 38.7 +/- 4.5 hours (p = 0.039), respectively. CONCLUSIONS: Mandatory surgical intensive care unit admission of all patients after radical cystectomy and urinary diversion does not appear indicated. A subset of patients at low risk for requiring active treatment may be identified who may be safely treated in an intermediate care setting after initial postoperative observation in the recovery room. The results of our retrospective analysis and risk stratification model should be validated in a prospective trial. PMID- 11435855 TI - The laparoscopic cigarette sponge. AB - PURPOSE: Minor hemorrhage during laparoscopic procedures may obscure the operative field. We describe the use of an especially designed, 4 x 4 absorbent sponge for multiple laparoscopic applications. MATERIALS AND METHODS: The cigarette sponge, also known as the Kittner roll gauze, was routinely used for laparoscopic upper tract procedures. The sponge may be placed easily through ports 5 mm. or greater. RESULTS: The cigarette sponge was excellent for absorbing minor but bothersome bleeding, facilitating suction and blunt dissection, and assisting with retraction. CONCLUSIONS: This especially designed laparoscopic sponge dramatically eases laparoscopic procedures, especially for controlling bothersome hemorrhage and blunt dissection. It may decrease operative time and facilitate difficult laparoscopic procedures. PMID- 11435856 TI - The pre-looped intracorporeal knot: a new technique for knot tying in laparoscopic surgery. AB - PURPOSE: Intracorporeal knot tying in laparoscopic surgery is time-consuming and difficult to learn. We present a new technique, that we call the pre-looped intracorporeal knot technique, which obviates the difficulties and saves time. MATERIALS AND METHODS: We devised a homemade suture ring that allows the introduction of the suture thread wrapped on the needle driver and ready for knot tying simply by pulling on both extremities of the thread. RESULTS: Our experience with this technique proved it to be easy to apply and to learn. CONCLUSIONS: The prelooped intracorporeal knot technique allies the sophistication of intracorporeal knot tying to the easiness and simplicity of the extracorporeal classic suturing. It renders intracorporeal knotting an easy and rapid task to achieve. PMID- 11435857 TI - An articulating retractor holder to facilitate laparoscopic adrenalectomy and nephrectomy. AB - PURPOSE: Minimally invasive surgical approaches to renal and adrenal tumors have gained increasing use. The addition of robotic assistance and mechanical devices has decreased the number of assistants required for these often long cases. We describe an articulating arm retractor holder to aid in liver and spleen retraction during laparoscopic surgery. MATERIALS AND METHODS: The articulating retractor holder consists of 4 components, including a base rod, flexible extension arm, stainless steel precision clamp and table attachment. During laparoscopy the abdomen is visualized using standard approaches. The articulating retractor holder is clamped to the table via the base rod and brought into the surgical field using sterile technique. A retractor is placed intracorporeally to elevate the liver or spleen and the handle is secured in place using the precision clamp. RESULTS: The articulating retractor holder was used in all right and approximately 50% of left laparoscopic cases. Adequate visualization was obtained in all patients. There were no device failures or retractor related complications. CONCLUSIONS: The articulating retractor holder is a useful tool to aid in laparoscopic retraction that should be added to the urologist armamentarium. PMID- 11435858 TI - Robot assisted laparoscopic nephrectomy. AB - PURPOSE: The feasibility and safety of remote laparoscopic surgery using a surgical telemanipulator have been demonstrated in laboratory experience and recently in clinical practice. To our knowledge we report the first robot assisted, laparoscopic nephrectomy in a human. MATERIALS AND METHODS: A 77-year old woman was diagnosed with a nonfunctioning hydronephrotic right kidney due to ureteropelvic junction obstruction. Robot assisted, transperitoneal right laparoscopic nephrectomy was performed. RESULTS: Complete dissection was successfully performed with the robot. The renal pedicle was dissected without any problem, and the artery and vein were individually ligated. Operative time was 200 minutes, anesthesia time was 245 minutes and blood loss was less than 100 ml. Convalescence was uneventful. Histological examination confirmed the preoperative diagnosis. CONCLUSIONS: We report the technical feasibility of robot assisted laparoscopic nephrectomy in humans. Current technology needs further improvement and its actual usefulness for patient treatment must be established by large clinical trials. Technological improvements and future telecommunication networks should open new avenues in surgery, namely remote telesurgery. PMID- 11435859 TI - Laparoscopic repair of renal artery aneurysm. AB - PURPOSE: We describe technical considerations of the laparoscopic repair of a renal artery aneurysm. MATERIALS AND METHODS: A 57-year-old woman presented with a 3 cm. aneurysm of the distal left main renal artery at its bifurcation. Using a purely laparoscopic 4-port transperitoneal technique the aneurysm was completely mobilized from its location behind the renal vein. Its 3 feeding vessels were controlled individually with bulldog clamps. The aneurysm sac was bivalved and precisely trimmed to conform with the diameter of the main renal artery. Vascular reconstruction was performed with running freehand laparoscopic suturing and intracorporeal knot tying using 4-zero polypropylene suture. RESULTS: Warm ischemia time was 31 minutes, total operative time was 4.2 hours, blood loss was 100 cc and hospital stay was 2 days. Postoperatively renal scan showed improved perfusion and renal arteriography confirmed adequate repair of the aneurysm. CONCLUSIONS: Laparoscopic repair of the renal artery aneurysm is feasible. To our knowledge we present the initial clinical report of laparoscopic renovascular surgery in the literature. PMID- 11435860 TI - Simplified ureteral stent placement with the assistance of a ureteral access sheath. AB - PURPOSE: When using a ureteral access sheath following a ureteroscopic procedure, placement of an internal ureteral stent can be simplified by inserting the stent through the sheath without the need to reinsert the cystoscope. MATERIALS AND METHODS: An indwelling ureteral stent with the pull string attached is inserted over the guide wire into the access sheath followed by the pusher. The guide wire is partially withdrawn allowing the stent to form a coil in the renal pelvis, using the pull string to adjust the stent position. The fluoroscopy unit is then focused onto the bladder and the guide wire is slowly withdrawn until its tip is at the level of pubic symphysis. The pusher and guide wire are then removed and the pull string is cut at the urethral meatus. RESULTS: Among 71 cases studied 60 required ureteral stent placement. In 43 of the 60 cases (72%) the ureteral access sheath greatly facilitated ureteroscopy, and a stent was placed through the access sheath in 34 (79%). Stent placement through the access sheath was successful in all cases, with an average time saving of 2.3 minutes per case, compared to placing the stent by reinserting a cystoscope. CONCLUSIONS: If an access sheath has already been placed during a ureteroscopic procedure and stent insertion is deemed necessary, the stent can be easily placed through the access sheath under fluoroscopic guidance without the need to reinsert the cystoscope. Our experience suggests that all urologists who routinely perform ureteroscopic procedures can easily master this timesaving technique. PMID- 11435861 TI - Percutaneous nephrolithotomy of a pelvic kidney: a posterior approach through the greater sciatic foramen. PMID- 11435862 TI - Gonadotropin-releasing hormone agonist failure in a man with prostate cancer. PMID- 11435863 TI - Perineal prostatic cancer seeding following radioactive seed brachytherapy. PMID- 11435864 TI - High dose hook effect in serum total and free prostate specific antigen in a patient with metastatic prostate cancer. PMID- 11435865 TI - Followup of primary malignant melanoma of the prostate. PMID- 11435866 TI - Penile necrosis secondary to spontaneous penile torsion. PMID- 11435867 TI - Ureteral obstruction due to colonic duplication. PMID- 11435868 TI - Acute urinary retention due to benign prostatic hyperplasia in a 23-year-old patient. PMID- 11435869 TI - Re: Higher urinary potassium is associated with decreased stone growth after shock wave lithotripsy. PMID- 11435870 TI - Re: Cable tie compression to facilitate laparoscopic partial nephrectomy. PMID- 11435871 TI - Re: Mucin gene expression in human urothelium and in intestinal segments transposed into the urinary tract. PMID- 11435872 TI - Re: Placenta percreta with bladder invasion as a cause of life threatening hemorrhage. PMID- 11435873 TI - Short-term changes in renal function after extracorporeal shock wave lithotripsy in children. AB - PURPOSE: To our knowledge despite numerous studies the biological effect of extracorporeal shock wave lithotripsy (ESWL, Dornier Medical Systems, Inc., Marietta, Georgia) on the function of the immature kidney has not yet been evaluated. We determined the short-term effect of ESWL on renal function in children. MATERIALS AND METHODS: In a 5-year period 65 children were treated for 77 kidney stones by ESWL and followed regularly at our department. In 16 children the short-term effect of shock wave energy was studied by measuring blood parameters (sodium, potassium, urea, creatinine and C-reactive protein), urinary electrolytes (sodium, potassium and creatinine), urinary enzyme activity (aspartate transaminase, alanine transferase, alkaline phosphatase and lactate dehydrogenase) and the excretion of beta 2-microglobulin. Samples were obtained immediately before and 2 hours after ESWL, and on days 1, 2, 8, 15, 30 and 90 after treatment. RESULTS: Morphological changes in the kidneys were not detected by ultrasound during followup. No significant changes were noted in overall renal function, serum parameters or urine electrolytes. A significant elevation in the excretion of aspartate transaminase, alkaline phosphatase, lactate dehydrogenase and beta 2-microglobulin was observed, indicating proximal tubular dysfunction and cell destruction. Enzyme levels returned to baseline within 15 days. CONCLUSIONS: Our results demonstrate that shock wave energy induces transient functional damage of tubular function in children. Minimizing the kV. and number of shocks may decrease the deleterious effect. When considering functional regeneration time, the minimal interval between 2 shock wave treatments should be at least 15 days. The long-term effect needs further investigation. PMID- 11435874 TI - Penile anatomy under the pubic arch: reconstructive implications. AB - PURPOSE: We have previously defined the anatomy of the neurovascular bundle in the normal and hypospadiac penis. These studies were based on analysis of the fetal penis distal to the pubic arch without total inclusion of the crural bodies. To our knowledge the neuroanatomy beneath the pubic arch has not been well described. We defined the nerve distribution under the pubic arch and the relationship of the nerves to the crural bodies, corporeal bodies and urethra of the penis. MATERIALS AND METHODS: Eight normal human fetal penile specimens (at 17.5 to 29 weeks of gestation and 1 hypospadiac specimen at 32 weeks were serially sectioned and stained with Masson's trichrome, and the neuronal markers protein gene product 9.5 and S-100. These specimens were unique in that they contained the whole penis from the glans to the crural bodies beneath the pubic arch. Older specimens were decalcified before fixation. Computer reconstruction with commercially available graphics software allowed 3-dimensional analysis of the nerves and crural bodies in relation to the pubic arch and surrounding structures. RESULTS: The nerves of the penile shaft and glans surrounded the corporeal bodies, extending from the junction of the urethral spongiosum to the classic 11 and 1 o'clock positions with a paucity of nerves at the 12 o'clock position in the dorsal midline. Beneath the pubic arch the nerves to the penis were an extension of the dorsal neurovascular bundle of the prostate. The nerves formed 2 bundles following a path just under the pubic arch in close proximity to the bone, superior to the urethra and medial to the origin of the crural bodies. The nerve bundles joined the corporeal bodies at the proximal origin, where the 2 crural bodies fused together. At this point perforating branches into the corporeal bodies from the cavernous nerves were documented. As the dorsal nerves joined the dorsal aspect of the corporeal bodies, they immediately began to fan out along the surface of the corporeal tissue to the junction of the urethral spongiosum. Three-dimensional reconstruction showed the relationship of the nerves to the pubic arch and urethra in multiple views. CONCLUSIONS: A precise understanding of penile anatomy beneath the pubic arch and at the origin of the crural bodies is important for preserving neuronal structures. This anatomy is especially germane in children undergoing posterior urethral reconstruction secondary to trauma, intersex requiring feminizing genitoplasty and severe hypospadias. PMID- 11435875 TI - The natural history of idiopathic urethrorrhagia in boys. AB - PURPOSE: We describe the presenting symptoms, evaluation and natural history of urethrorrhagia in boys. MATERIALS AND METHODS: The records of 27 consecutive toilet trained boys with idiopathic urethrorrhagia were retrospectively reviewed for information regarding age, symptoms, symptom duration, physical examination, and radiographic, endoscopic and laboratory data. Patient interviews were performed, and the resolution rate, symptom duration and associated urological abnormalities were evaluated. RESULTS: Mean age at presentation was 10.1 years. Symptoms included urethrorrhagia in 100% and dysuria in 29.6% of cases. Radiographic and laboratory evaluations were normal in all patients except for microscopic hematuria in 57%. Cystourethroscopy in 4 patients revealed bulbar urethral inflammation without stricture in 2. A total of 24 patients (89%) were followed an average of 37 months (range 10 to 106). Complete resolution developed in 46% of cases at 6 months, in 71% at 1 year and in 91.7% overall. The average duration of symptoms was 9.9 months (range 2 weeks to 38 months). In 2 boys (8.3%) urethrorrhagia persisted for 22 and 28 months, and in 1 cystoscopy revealed urethral stricture. Self-limiting urethrorrhagia recurred in 2 patients (8.3%) after initial resolution. Treatment consisted of watchful waiting in all patients except 1 with urethral stricture, who underwent urethral dilation. After urethrorrhagia resolved epididymo-orchitis recurred in 1 case. CONCLUSIONS: Routine radiographic, laboratory and endoscopic evaluation is unnecessary for evaluating urethrorrhagia. Watchful waiting is indicated because the condition resolves in 71% and 91.7% of patients at 1 and 2 years, respectively. Evaluation should be considered in patients with prolonged urethrorrhagia because urethral stricture may be identified. PMID- 11435876 TI - Dismembered V-flap pyeloplasty. AB - PURPOSE: We present a modified technique of pyeloplasty that seems ideally suited for reoperative as well as primary repair of ureteropelvic junction obstruction due to high ureteral insertion. MATERIALS AND METHODS: This repair combines the dismembered technique with creation of a V-flap of renal pelvis by dividing and incising through the ureteropelvic junction superolateral onto the pelvis to a height above the most dependent portion of the pelvis just exceeding the length of ureteral spatulation. The V-flap is completed by an inferolateral incision directed toward the lateral aspect of the pelvis at its most dependent portion. The tip of the V-flap is then flipped down and approximated to the apex of the posterior ureteral spatulation. Of the 12 cases managed by this procedure 2 were reoperative and 10 involved select primary repair with high ureteral insertion at the ureteropelvic junction. Patient age was 3 months to 17 years (median 11 months). Two procedures were performed via a dorsal lumbar incision in the oldest patients and 10 were done via the standard anterior extraperitoneal approach. Radiological evaluation, including mercaptoacetyltriglycine renal scan and/or excretory urography, was performed in all patients preoperatively and postoperatively. RESULTS: In all patients postoperative mercaptoacetyltriglycine renal scan and/or excretory urography at 3 months and 1 year showed excellent drainage after dismembered V-flap pyeloplasty. CONCLUSIONS: Dismembered V-flap pyeloplasty has proved to be successful with a number of advantages over the Anderson-Hynes dismembered technique in select patients. These advantages include the avoidance of tissue tension that make mobilization of the kidney and ureter unnecessary, aggressive ureteral spatulation, creation of a dependent, funneled configuration and automatic tapering of a redundant pelvis in the routine course of closure without excision of tissue. This technique also has the advantage of being simpler than other flap repairs and it combines the physiological virtues of dismembered repair with the anatomical advantages of flap pyeloplasty. PMID- 11435877 TI - No relationship of testicular size at orchiopexy with fertility in men who previously had unilateral cryptorchidism. AB - PURPOSE: We determined whether testicular size at orchiopexy is predictive of fertility potential and whether size correlates with sperm parameters, hormone levels or testicular volume in adulthood. MATERIALS AND METHODS: Testicular size obtained from the operative notes of patients who underwent unilateral orchiopexy was classified as normal, small or large for age. These data were compared with outcome data. Paternity data were included on 166 men who reported achieving or unsuccessfully attempting paternity for 12 or more months. Of the men 49 provided blood for hormone level measurement, 43 underwent semen analysis and in 29 testicular volume was determined. RESULTS: In the 166 men, including 98, 65 and 3 with small, normal and large testes for age at orchiopexy, respectively, there was no difference in paternity based on testicular size. Of those who achieved paternity time to conception did not differ based on size. Mean age at surgery also did not differ, nor did the percent of small versus normal testes in the age categories 0 to 2, 3 to 5, 6 to 8 and 9 to 11 years. There was no difference in men with small or normal testicular size at surgery in mean luteinizing hormone, follicle-stimulating hormone, testosterone or inhibin B. Sperm density, motility and morphology, and volume in adulthood of a previously undescended testis, previously descended testis or previously undescended and descended testes did not differ in these 2 groups, although the previously undescended testis was smaller than the contralateral testis. CONCLUSIONS: In men with a history of unilateral cryptorchidism small testicular size at orchiopexy is not associated with decreased paternity (89.8%), abnormal hormone levels, a lower sperm count or decreased testicular volume in adulthood. PMID- 11435878 TI - Parental preferences in the management of vesicoureteral reflux. AB - PURPOSE: We determined parental preferences for the treatment of vesicoureteral reflux in their child. MATERIALS AND METHODS: Parents of children with vesicoureteral reflux were prospectively recruited to evaluate choices in reflux management. In each case a standard questionnaire that described the treatment options for reflux was administered. Parents were asked to choose between long term antibacterial prophylaxis with annual radiography studies and open or endoscopic treatment at each of 1 to 5 years of followup. They were also given the choice between open or endoscopic treatment. Annual resolution and/or correction rates provided for medical, surgical and endoscopic management were 20%, 95% to 100% and 80% after 1 or 2 injections, respectively. RESULTS: We queried 91 families of female (81%) and male (19%) patients. Average duration of reflux followup was 2 years and mean patient age was 49.8 months. At diagnosis reflux was grades I to II in 65% of cases, grade III in 26% and grades IV to V in 9%. The majority of parents chose daily antibiotics over surgery if the child was predicted to have vesicoureteral reflux for 1 to 4 years. However, the majority chose ureteral reimplantation over daily antibiotics and yearly x-ray if a 5-year course was predicted. In contrast, parents chose daily antibiotics rather than endoscopic treatment if the anticipated interval was 1 to 3 years. After 3 years the majority preferred the endoscopic approach. Also, 60% of parents stated that they would choose endoscopic treatment over reimplantation, although the child may require repeat endoscopic treatment and there was a 20% chance of persistent vesicoureteral reflux. CONCLUSIONS: Parents of children with vesicoureteral reflux prefer antibiotic prophylaxis as initial treatment. However, when daily antibiotics and yearly cystography may be required beyond 3 to 4 years, most parents would choose definitive correction. While endoscopic treatment is less effective than surgery, parents prefer endoscopic treatment, most likely because it is less invasive. Also, when compared directly against each other, the majority of parents stated that they would choose endoscopic treatment over surgery, although it has a lower success rate. PMID- 11435879 TI - The logic and limits of shared decision making. PMID- 11435880 TI - Gastrocystoplasty and chronic renal failure: an acid-base metabolism study. AB - PURPOSE: To verify in an experimental model whether gastrocystoplasty may protect patients with chronic renal failure from acid loading associated acidosis a wedge shaped portion of the middle stomach was used to improve bladder capacity in animals with chronic renal failure. MATERIALS AND METHODS: An experimental model was used to study 112 adult female Wistar rats (EPM-1) weighing between 156 and 259 gm. The animals were randomly assigned to groups, including 41 controls, 24 undergoing five-sixths nephrectomy to create chronic renal failure, 26 undergoing gastrocystoplasty and 21 undergoing gastrocystoplasty and five-sixths nephrectomy to create renal failure. To provide an acid overload a 5% NH(4)Cl diet was administered to a subgroup of each group. Two months after surgery 24-hour urine was collected, and volume and pH were measured as well as the amount of bicarbonate, ammonium, titratable acidity and chloride. The animals were then exsanguinated through an abdominal aorta puncture. The blood was used for blood gas analysis and to measure sodium, potassium, chloride, ionized calcium and creatinine. RESULTS: When undergoing an acid overload, the animals with gastrocystoplasty had no acidosis since acid radicals were eliminating in the urine as NH(4)Cl. When given the same acid overload, metabolic acidosis developed in the animals with gastrocystoplasty and chronic renal failure. CONCLUSIONS: Gastrocystoplasty protected healthy rats from acidosis when they were given an acid overload but failed to protect the rats with chronic renal failure under the same conditions. PMID- 11435881 TI - Demonstration of neuronal networks in the human upper urinary tract using confocal laser scanning microscopy. AB - PURPOSE: To our knowledge innervation of the upper urinary tract and its role in motility and sensation are not clearly understood. The whole mount preparation technique provides 3-dimensional (D) morphology of the innervation and its relationship of branching and interconnecting nerve fibers to each other and to the neighboring tissues. Confocal laser scanning microscopy provides dramatic optical advantages for detecting 3-D structures in thick specimens. We investigated the distribution and morphology of the neuronal structures in the human upper urinary tract using the whole mount preparation technique and confocal laser scanning microscopy. MATERIALS AND METHODS: Whole mount preparations of the human renal pelvis and ureter were stained by standard immunohistochemical method using various neuronal markers (protein gene product 9.5, neuron specific enolase and neurofilament). The 3-D architecture of the specimens was investigated with the help of confocal laser scanning microscopy. RESULTS: We detected 2 mesh-like neuronal networks or plexus in the human upper urinary tract. The first and more prominent plexus was located in the submucosa between the lamina propria and tunica muscularis, and the second neuronal network was found between the smooth muscle fibers of the ureteral wall. There were frequent interconnections between the 2 networks in the ureteral wall. CONCLUSIONS: To our knowledge our study shows for the first time that there are 2 well formed mesh-like neuronal plexus in the human upper urinary tract. Our findings suggest that the autonomic nervous system of the human upper urinary tract may have a significant role in the propagation, coordination and modulation of ureteropelvic peristalsis. PMID- 11435882 TI - Bladder transplantation in rats using FK-506. AB - PURPOSE: We created viable bladder tissue by transplantation with immunosuppression. MATERIALS AND METHODS: For bladder transplantation the bladder of newborn Brown-Norway rats was excised and each was transplanted into a pouch created in the distal omentum of a 5-week-old Lewis rat. In 15 group 1 rats no immunosuppressive agent was used. In 20 group 2 rats 0.6 mg./kg. FK-506 daily were given intramuscularly until a predetermined day of harvest. Recipient rats were sacrificed on day 3, 5, 7 or 14 after bladder transplantation, and the bladder grafts were harvested and formalin fixed. Hematoxylin and eosin staining was done to examine bladder graft survival and the degree of rejection, and immunohistochemical testing was performed for assessing the vesical nervous system. In 5 rats in the control group bladder augmentation was performed by anastomosing the bladder graft to the native bladder. Each augmented bladder was harvested 21 days later for histopathological assessment. RESULTS: Overall bladder graft survival was 96.4%. Each successfully transplanted bladder graft appeared macroscopically as a thin walled cyst. In group 1 all bladder grafts showed rejection with cellular infiltration. In group 2 there was mild rejection in 5 rats and no evidence of rejection in the remaining 15. All group 2 bladder grafts had intact nerve distribution. Bladder augmentation was successful in all 5 cases and the mucosa was normal throughout each augmented bladder. CONCLUSION: Because FK-506 successfully prevents rejection, our technique would appear to have the potential for creating viable bladder tissue that may be used for bladder augmentation in cases of vesical exstrophy or neurogenic bladder. PMID- 11435883 TI - Effect of grapefruit juice on urinary lithogenicity. AB - PURPOSE: An increased risk of nephrolithiasis has been associated with the ingestion of grapefruit juice in epidemiological studies. To our knowledge the basis of this effect of grapefruit juice has not been studied previously. We studied the effect of grapefruit juice consumption on urinary chemistry and measures of lithogenicity. MATERIALS AND METHODS: Ten healthy men and women between ages of 25 and 40 years participated. Each subject drank 240 ml. of tap water at least 3 times daily for 7 days during the control period. This period was followed by a second 7 days experimental period during which they drank 240 ml. of grapefruit juice 3 times daily. In each 7-day period urine was collected for 24 hours during the last 3 days. Urine chemical analysis was performed, supersaturations of calcium oxalate, calcium phosphate and uric acid were calculated and urinary lithogenicity was measured. RESULTS: Urine volume and creatinine excretion were the same during the control and experimental periods. Grapefruit juice ingestion was associated with an increase in mean oxalate excretion plus or minus standard deviation of 41.1 +/- 9.2 to 51.9 +/- 12.0 mg. per 24 hours (p = 0.001) and in mean citrate excretion of 504.8 +/- 226.5 to 591.4 +/- 220.0 mg. per 24 hours (p = 0.01). There was no net change in the supersaturation or upper limit of metastability of calcium oxalate, calcium phosphate or uric acid. Crystal aggregation and growth inhibition by urinary macromolecules was not affected by grapefruit juice ingestion. CONCLUSIONS: Offsetting changes in urine chemistry caused by the ingestion of grapefruit juice led to no net change in calculated supersaturation. No changes in lithogenicity were demonstrated. The results do not demonstrate an effect of grapefruit juice for increasing lithogenicity. The basis of the observations of epidemiological studies remain unexplained. PMID- 11435884 TI - Excretion of bikunin and its fragments in the urine of patients with renal stones. AB - PURPOSE: Bikunin is a Kunitz-type protease inhibitor found in serum and urine. It has been implicated in urinary stone formation. This study was designed to investigate the role of urinary bikunin in stone formation. MATERIALS AND METHODS: Urinary concentrations of bikunin were measured in 18 male formers of urinary stones 28 to 74 years old and in 77 healthy controls, including 39 males and 38 females, without urological abnormality. A sensitive competitive solid phase enzyme immunoassay was established for urinary bikunin. Bikunin was also qualitatively assessed by Western blot analysis. RESULTS: The mean urinary bikunin-to-creatinine ratio plus or minus standard deviation in stone formers was significantly elevated compared with that in healthy male and female controls (52.9 +/- 46.0 microg./mg. creatinine versus 28.0 +/- 30.4 and 26.5 +/- 21.7, p = 0.005 and 0.006, respectively). By Western blot analysis all urine samples contained authentic 40 kDa. bikunin species. However, a significantly higher proportion of patients was found to have aberrant 25 kDa. bikunin species compared with controls (10 of 18 or 55.6% versus 15 of 77 or 19.5%, p = 0.002). Experiments on de-glycosylation with chondroitinase ABC, amino acid sequencing of the aberrant bikunin species and calcium oxalate crystal growth inhibition assay demonstrated that the 25 kDa. bikunin fragment was identical to de-glycosylated bikunin and less inhibitory on calcium oxalate crystal growth. CONCLUSIONS: If urinary bikunin is important in the pathogenesis of urolithiasis, its effect is probably attributable to the concentration and degree of glycosylation. PMID- 11435885 TI - Protective role of enalapril for chronic tubulointerstitial lesions of hyperoxaluria. AB - PURPOSE: Hyperoxaluria is a recognized cause of tubulointerstitial lesions and it may contribute to chronic renal failure. In previous studies we demonstrated that enalapril was effective against the progression of tubulointerstitial lesions in a 4-week hyperoxaluria rat model. We evaluated whether the action of enalapril on the tubulointerstitial lesions produced by hyperoxaluria persisted for a long period. MATERIALS AND METHODS: Two-month-old male Sprague-Dawley rats were divided into 4 groups of 12 each, including 1--control animals given tap water, 2 -animals with hyperoxaluria, 3--animals with hyperoxaluria plus enalapril, 4- animals with enalapril. Hyperoxaluria in groups 2 and 3 rats was induced by administering 1% ethylene glycol, a precursor for oxalates, in the tap water continuously throughout the whole study. Meanwhile, groups 3 and 4 received 20 mg./l. enalapril in the drinking water. At the end of the study renal tubulointerstitial lesions were evaluated by immunostaining using monoclonal antibodies against macrophage infiltrates (ED1), tubulointerstitial alpha-smooth muscle actin and transforming growth factor-beta1. The lesions were quantified by semiquantitative scores. Creatinine clearance and urinary albumin excretion were also determined. RESULTS: There was no difference in urine oxalate excretion in groups 2 and 3. Group 3 rats treated with enalapril showed fewer tubulointerstitial lesions than nontreated group 2 rats, as indicated by the mean scores plus or minus standard error of mean for inflammatory infiltrate (2.16 +/- 0.2 versus 0.83 +/- 0.16), tubular atrophy (2 +/- 0.27 versus 0.66 +/- 0.14), interstitial fibrosis (2.5 +/- 0.15 versus 0.5 +/- 0.1), glomerular ED1 (1.75 +/- 0.25 versus 0.16 +/- 0.11), interstitial ED1 (2.33 +/- 0.18 versus 0.58 +/- 0.10) tubular transforming growth factor-beta1 (2.09 +/- 0.08 versus 0.91 +/- 0.14), interstitial transforming growth factor-beta 1 (2.33 +/- 0.22 versus 0.66 +/- 0.12), tubulointerstitial alpha-smooth muscle actin (2.91 +/- 0.22 versus 0.83 +/ 0.16), lower urinary albumin excretion (35.5 +/- 2.7 mg. daily versus 10.9 +/- 1) and higher creatinine clearance (2.29 +/- 0.04 ml. per minute versus 2.54 +/- 0.03, all p <0.05). CONCLUSIONS: Based on our results we believe that enalapril would provide a beneficial effect against chronic tubulointerstitial lesions caused by oxalates. PMID- 11435886 TI - Laparoscopic radiofrequency thermal ablation of renal tissue with and without hilar occlusion. AB - PURPOSE: Few studies have evaluated the effect of radiofrequency thermal ablation on renal tissue, although it has been used clinically to treat small renal masses. We studied the size and histology of lesions created with radiofrequency thermal ablation administered via the laparoscopic approach with and without hilar occlusion in a porcine model. MATERIALS AND METHODS: The lower pole of each kidney was exposed laparoscopically in 11 farm pigs. In each kidney a 7-electrode dry radiofrequency thermal ablation probe was inserted at an identical location and deployed to a diameter of 2 cm. Energy was applied for 8 minutes at an average temperature of 100C. The left renal hilum of each pig was clamped during radiofrequency thermal ablation. Two pigs were sacrificed immediately, and 3 each were sacrificed at 24 hours, 2 and 4 weeks. The size and shape of the lesions created were measured and examined histologically. RESULTS: There were no intraoperative or postoperative complications. Laparoscopic ultrasound confirmed probe placement but did not monitor lesion progression. Acutely lesions were firm and white with a small adjacent hemorrhagic zone. Histological evaluation revealed preserved renal architecture but the loss of distinct cytoplasmic features. Nicotinamide adenine dinucleotide staining demonstrated no viable cells within the lesions. In surviving animals pelvicaliceal integrity was preserved. In the 2 and 4-week survival groups kidneys treated with hilar occlusion had larger lesions than nonoccluded kidneys but the differences were not significant at 4 weeks (3.2 x 2.7 x 2.5 cm. and 3.5 x 1.7 x 2.0, respectively, p >0.05). Histologically untreated parenchyma of hilar occluded kidneys demonstrated changes consistent with chronic pyelonephritis. In 1 kidney radiofrequency thermal ablation with hilar occlusion resulted in complete lower pole loss at 4 weeks. CONCLUSIONS: In the porcine model renal radiofrequency thermal ablation creates rapid and completely devitalized lesions of consistent size and shape. Hilar occlusion may result in slightly larger lesions but risks damage to the whole renal unit. PMID- 11435887 TI - Laparoscopic intracorporeally constructed ileal conduit after porcine cystoprostatectomy. AB - PURPOSE: We present our technique of laparoscopic ileal conduit creation after cystoprostatectomy in a porcine model performed in a completely intracorporeal manner. METHODS AND METHODS: After developing the technique in 5 acute animals laparoscopic cystoprostatectomy with intracorporeally performed ileal conduit urinary diversion was performed in 10 surviving male pigs. A 5-port transperitoneal technique was used. All steps of the technique applied during open surgery were duplicated intracorporeally. Specifically cystectomy, isolation of an ileal conduit, restoration of bowel continuity and mucosa-to-mucosa stented bilateral ileoureteral anastomosis formation were performed by exclusively intracorporeal laparoscopic techniques. RESULTS: Surgery was successful in all 10 study animals without intraoperative or immediate postoperative complications. Blood loss was minimal and average operative time was 200 minutes. Stenosis of the end ileal stoma specifically at the skin level was noted in 6 animals. Three deaths occurred 2 to 3 weeks postoperatively. At sacrifice renal function was normal in all surviving animals. No ileo-ureteral anastomotic strictures were noted on pre-sacrifice radiography of the loop or at autopsy examination of the anastomotic sites. CONCLUSIONS: Laparoscopic ileal conduit urinary diversion after cystoprostatectomy may be performed completely intracorporeally in the porcine model. Clinical application of this technique is imminent. PMID- 11435888 TI - Percutaneous cryoablation of porcine kidneys with magnetic resonance imaging monitoring. AB - PURPOSE: We determined the feasibility of a percutaneous approach using magnetic resonance imaging (MRI) for creating cryoablation lesions in the porcine kidney. METHODS AND METHODS: Three domestic swine underwent renal cryoablation under MRI guidance with a total of 6 cryoablation lesions created in 5 kidneys. A 3 mm. cryoprobe was placed under MRI guidance using an interventional MRI unit. With a pressurized argon gas cooling unit the cryoablation lesion was created and monitored by MRI. Gross and histological examination of the kidneys was performed 1 week after the procedure. RESULTS: All animals survived the procedure without difficulty. A total of 6 cryoablation lesions were produced in 5 kidneys. The lesions were 1.9 x 1.3 to 3.9 x 1.9 cm. on MRI. Histological examination 1 week after treatment showed that the lesions were 1.7 x 1.0 to 3.2 x 1.2 cm. There was an area of coagulation necrosis surrounded by a transition zone of inflammatory reaction a mean of 0.5 cm. in diameter with each lesion. CONCLUSIONS: Percutaneous renal cryoablation using MRI imaging proved to be a successful technique for guiding probe placement and monitoring ice ball formation. Because MRI allows imaging in 3 planes, it may be useful for cryoablation of intraparenchymatous tumors. PMID- 11435889 TI - Development of a radiofrequency based thermal therapy technique in an in vivo porcine model for the treatment of small renal masses. AB - PURPOSE: Incidentally detected small renal tumors appear to grow slowly and be localized to the kidney. Minimally invasive therapies are being investigated as alternatives to standard surgical techniques. Radiofrequency ablation has been reported for the treatment of small renal cell carcinomas. We developed a radiofrequency technique and established its efficacy and safety in a large animal model. METHODS AND METHODS: A total of 22 lesions were created in normal kidneys of 7 pigs. Radiofrequency energy was administered during open exposure of the kidneys or percutaneously under ultrasound guidance. Lesion development was monitored with gray-scale and power Doppler ultrasound. To avoid heating surrounding tissues new hydro-dissection and gas-dissection techniques were developed. Lesion sizes and characteristics were assessed by ultrasound and pathological examination. RESULTS: No complications were observed due to probe insertion and removal. Perirenal structures were thermally damaged before the development and application of the dissection techniques. Lesion size was accurately predicted by gray-scale ultrasound on day 7. Loss of perfusion in the ablated volume was confirmed by power Doppler ultrasound. Lesions were wedge shaped, presumably due to the effects of heating on segmental blood flow distribution. Pathological examination revealed changes consistent with thermal injury and ischemic type infarction. CONCLUSIONS: Radiofrequency thermal therapy is an effective and efficient method for ablating normal renal tissue in the pig. It may be applied percutaneously under ultrasound guidance with minimal complications provided that vital adjacent structures are protected from thermal damage. Further studies are required in humans before adopting this technique as definitive treatment for small renal cell carcinoma. PMID- 11435890 TI - T cells activated in vitro as immunotherapy for renal cell carcinoma: characterization of 2 effector T-cell populations. AB - PURPOSE: Effector T cell populations generated using 2 methods of in vitro activation are currently being tested in separate clinical trials as immunotherapy for patients with advanced cancer, including renal cell carcinoma. To determine the most appropriate method of activation for cancer immunotherapy in vitro antitumor activity of the 2 effector T-cell populations were compared. METHODS AND METHODS: The effector T-cell populations were generated concurrently by activation of peripheral blood mononuclear cells from patients with advanced renal cell carcinoma or other cancer using soluble anti-CD3 monoclonal antibody (3T cells) or anti-CD3 and anti-CD28 monoclonal antibodies immobilized on beads (3/28T cells). After 14-day culture the phenotype and functional activity of the cells were tested. RESULTS: Fold expansion of CD4(+) cells for 3T cultures was lower than for 3/28T cultures but expansion of CD8(+) cells was similar for both cultures. Expression of CD69 was higher on 3T cells. 3T and 3/28T cells exhibited similar ability to kill various human tumor cell lines. Although both effector T cell populations produced Th1-type cytokines upon re-stimulation, 3T cells secreted a higher level of interferon-gamma and tumor necrosis factor-alpha compared with 3/28T cells. Intracellular cytokine analysis demonstrated that the percent of cells producing interferon-gamma was higher in CD4(+), CD8(+), CD25(+), CD69(+) and CD45RO(+) 3T cells compared with 3/28T cells. CONCLUSIONS: These data suggest that 3T cells may have increased efficacy as immunotherapy for patients with cancer due to higher levels of tumoricidal cytokine production than 3/28T cells. PMID- 11435891 TI - Various forms of mutant p53 confer sensitivity to cisplatin and doxorubicin in bladder cancer cells. AB - PURPOSES: The nature of p53 mutation has been reported to affect cellular responses to chemotherapy. We characterized the impact of p53 mutations on drug resistance in bladder cancer cells. METHODS AND METHODS: Various human p53 mutants (V143A, V173L, H179Q, N247I and R273H) were introduced to the TCC-SUP bladder carcinoma cell line to establish stable transfectants. The expression of mutant p53 was demonstrated by reverse transcriptase-polymerase chain reaction and immunocytochemical analysis. The sensitivity to cisplatin and doxorubicin in these transfectants was determined by trypan blue exclusion. Cell death mediated by cisplatin and doxorubicin was characterized by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling analysis, Hoechst 33258 staining and annexin-V binding assay. RESULTS: The expression of all forms of mutant p53 protein except p53His273 enhanced sensitivity to cisplatin and doxorubicin. The chemosensitivity of p53His273 transfectants is similar to that of parental TCC-SUP and control transfectants. Cisplatin induced cell death undergoes apoptosis, as demonstrated by Hoechst staining, annexin-V assay and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling, respectively. In contrast, doxorubicin induced cell death probably occurs through a nonapoptotic pathway. CONCLUSIONS: These results indicated that the nature of p53 mutations may affect the cellular response to anticancer drugs and many forms of mutant p53 protein may enhance chemosensitivity through apoptotic or nonapoptotic pathways in bladder cancer cells. PMID- 11435892 TI - The effect of simulated birth trauma and/or ovariectomy on rodent continence mechanism. Part I: functional and structural change. AB - PURPOSE: We examined the changes in the lower urinary tract after delivery, intravaginal ballooning and/or ovariectomy. MATERIALS AND METHODS: The study included 10 virgin and 48 primiparous pregnant rats. Cystometry and the stress/sneeze test were performed in virgin and postpartum rats shortly after delivery and at 8 weeks before sacrifice. Half of the delivered animals underwent intravaginal balloon dilation. Four weeks later half in each group underwent ovariectomy. The rats were subdivided into group 1--delivery, group 2--delivery plus balloon inflation, group 3--delivery plus ovariectomy and group 4--delivery plus balloon inflation plus ovariectomy. Tissues from the bladder, bladder neck, urethra and levator were collected, analyzed by electron microscopy, and immunostained for caveolin-1, caveolin-3 and neuronal nitric oxide synthase. RESULTS: Higher bladder capacity was detected in postpartum than in virgin rats. Urine leakage on stress/sneeze testing increased significantly in groups 2 and 4. Electron microscopy revealed a significant decrease in sarcolemma caveolae in the smooth muscle of the bladder and urethra in groups 2 to 4. In the bladder neck in group 3 caveolae were increased in smooth muscle. In groups 2 to 4 in the smooth muscle of the bladder and urethra caveolin-1 was significantly decreased. Caveolin-3 and neuronal nitric oxide synthase in striated muscle also significantly decreased in groups 2 to 4. CONCLUSIONS: These findings suggest that birth trauma simulated by ballooning and ovariectomy may contribute to stress urinary incontinence. The alteration in smooth muscle caveolae as well as the membrane protein caveolin may have a role in functional alterations caused by birth trauma and ovariectomy. PMID- 11435893 TI - Involvement of hypogastric and pelvic nerves for conveying cystitis induced nociception in conscious rats. AB - PURPOSE: We determined the sites of the antinociceptive action of morphine in the experimental model of cyclophosphamide induced cystitis and investigated the afferent nerve fibers involved in nociception transmission originating from the bladder. MATERIALS AND METHODS: Cyclophosphamide (200 mg./kg.) given intraperitoneally was used to induce cystitis in male rats and their behavior was observed and scored. The effect of 2 mg./kg. systemic morphine given intravenously on cyclophosphamide induced behavioral modifications was tested when administered alone and after 100 microg. naloxone per rat given intrathecally at the L1 to L2 or L6 to S1 level. The spinal antinociceptive effect of morphine was also tested when administered intrathecally alone at 10, 100 and 200 microg. per rat at L1 to L2, alone at 100 microg. per rat at L1 to L2 or L6 to S1, alone at 100 microg. per rat at L1 to L2 and L6 to S1 simultaneously, alone at 200 microg. per rat at L1 to L2 and after 100 microg. naloxone per rat given intrathecally at L6 to S1 at 100 microg. per rat at L1 to L2. RESULTS: Cyclophosphamide induced marked modifications in the behavior of the rats, including a decreased breathing rate, eye closing and specific postures. Morphine given intravenously reversed these behavioral disorders and this reversal was completely prevented by pretreatment with intrathecal naloxone. A dose of 100 microg. per rat given intrathecally also reversed these behavioral disorders by about 25% at the L1 to L2 and L6 to S1 levels. In addition, a dose of 100 microg. morphine per rat administered intrathecally and simultaneously at L1 to L2 and L6 to S1 produced an effect equal to the sum of those observed when administered separately, that is about 50%, whereas morphine at an intrathecal dose of 200 microg. at L1 to L2 produced the same effect as 100 microg. given intrathecally at the same level or at L6 to S1 (25%). Also, 100 microg. naloxone per rat administered intrathecally at L6 to S1 prevented the effect of 100 microg. morphine at L1 to L2. CONCLUSIONS: These results confirm the previously reported antinociceptive effect of systemic morphine in this model of cyclophosphamide cystitis, suggest that this antinociceptive action is completely located at the spinal site and most importantly demonstrate by the pharmacological approach and behavioral analysis that nociceptive sensations originating from the bladder are conveyed by hypogastric and pelvic nerves in this cyclophosphamide cystitis model in the conscious rat. PMID- 11435894 TI - Cell relationship in a Wistar rat model of spontaneous prostatitis. AB - PURPOSE: Prostatitis in men is a painful, noninfectious inflammatory condition. It is similar to interstitial cystitis which is associated with increased bladder mast cell and sensory nerve fiber density as well as suprapubic pain. Certain strains of rats may provide a useful model for studies of the development of spontaneous prostatitis. We evaluated the time course, and involvement of mast cells and sensory nerve fibers in this process using Wistar rats. MATERIALS AND METHODS: The prostates of 4, 6, 8, 10 and 13-week-old male Wistar rats were examined for the degree of inflammation, innervation, mast cell density and nerve mast cell relationship using histochemical and immunocytochemical studies. Bacterial cultures of tissue were performed at 13 weeks. RESULTS: The inflammatory cell index increased progressively with age. Inflammation was moderate and consisted mostly of lymphocytes and macrophages associated with occasional glandular epithelial necrosis and edema. The density of nerve fibers immunoreacting with the neuronal marker protein gene produce 9.5 increased gradually with age and fibers immuno-positive for the sensory neuropeptide calcitonin gene-related peptide more than doubled by 13 weeks compared with by 4 weeks. The density of visible mast cells declined after 4 weeks in a pattern that corresponded with the increased percent of mast cells undergoing degranulation. For the mast cells with calcitonin gene-related peptide immuno-positive nerve fibers within a distance of 40 microm. distance correlated significantly with the degree of degranulation. Bacterial cultures were negative at 13 weeks. CONCLUSIONS: Our results confirm previous reports of spontaneous prostatitis in Wistar rats and indicate that moderate inflammation may occur in 80% of rats at as early as age 13 weeks. While the correlation of the nerve mast cell axis with mast cell degranulation does not prove our hypothesis of mast cell mediated inflammatory mediator release in the development of nonbacterial prostatitis, it suggests that such a relationship is possible. PMID- 11435895 TI - Characterization of the spontaneous electrical and contractile activity of smooth muscle cells in the rat upper urinary tract. AB - PURPOSE: We morphologically and electrophysiologically identified the cells that generate the electrical activity underlying the peristaltic contractions of the rat upper urinary tract. MATERIALS AND METHODS: Electron microscopy and tension recording techniques were used to characterize the smooth muscle cells underlying spontaneous contractions in the wall of the rat ureter, and proximal and distal renal pelvis. Intracellular microelectrodes, containing 4% neurobiotin were used to record data from the cells of the renal pelvis, which were later viewed on a confocal microscope. RESULTS: Spontaneous myogenic contractions (average 22.3 +/- 2.2 minutes(-1)) originated in the proximal renal pelvis and propagated into the distal renal pelvis and ureter in 6 preparations. Smooth muscle cells in the renal pelvis and ureter were typical in appearance with greater than 85% of their sectional area containing clumped contractile filaments. In contrast, contractile fibrils occupied only 65% of the sectional area of the smooth muscle cells within the most proximal region of the renal pelvis (pelvicaliceal junction). In strips of the renal pelvis spindle shaped cells 83 to 200 microm. long fired spontaneous action potentials (6 minutes(-1)) consisting of an initial spike, a quiescent plateau phase and abrupt hyperpolarization to a peak diastolic potential of -60 mV. Other spindle shaped cells 94 to 112 microm. long displayed small membrane transients (15 minutes(-1)) 9 to 19 mV. in amplitude, firing from a diastolic potential of -40 mV. CONCLUSIONS: It is likely that the spontaneous contractile activity of the rat upper urinary tract arises from the discharge of action potentials in typical smooth muscle cells of the proximal renal pelvis that are directly driven by the spontaneous membrane oscillations of atypical smooth muscle cells. PMID- 11435896 TI - Effects of isoproterenol on spontaneous excitations in detrusor smooth muscle cells of the guinea pig. AB - PURPOSE: Because beta-adrenoceptor agonists would be a useful tool for the pharmacological treatment of unstable bladder, we investigated the cellular mechanisms underlying beta-adrenoceptor mediated inhibition on spontaneous excitation in detrusor smooth muscle. MATERIALS AND METHODS: Detrusor smooth muscle bundles were isolated from guinea pig bladders. Changes in membrane potential were recorded using an intracellular recording technique. In preparations loaded with the calcium indicator fura-PE3 changes in the concentration of intracellular calcium ions were measured simultaneously with membrane potential. Effects of isoproterenol on spontaneous changes in the membrane potential and intracellular Ca(2+) were examined RESULTS: Detrusor smooth muscle cells exhibited spontaneous action potentials that were associated with transient increases in intracellular Ca(2+) (calcium transients). Isoproterenol, which hyperpolarized the membrane, prevented action potentials and calcium transients. This induced inhibition of calcium transients was not affected by cyclopiazonic acid. Isoproterenol induced hyperpolarization was inhibited by inhibitors of protein kinase A, N-[2-((p-bromocinnamyl)amino)ethyl] 5-isoquinolinesulfonamide, hydrochloride and Rp-adenosine-3',5'-cyclic phosphorothioate. Hyperpolarization was blocked by a solution containing 30 mM. potassium but not by a range of potassium channel blockers. Ouabain and a solution of 0.5 mM. potassium also inhibited hyperpolarization. CONCLUSIONS: Our results suggest that isoproterenol prevented spontaneous action potential discharges and associated calcium transients through the activation of protein kinase A. The isoproterenol induced inhibition of intracellular Ca(2+) largely depends on the prevention of spontaneous action potentials since the contribution of the intracellular calcium store was small. Isoproterenol hyperpolarizes the membrane, probably by stimulating sodium pump activity. PMID- 11435897 TI - Protective effect of vitamin E on the response of the rabbit bladder to partial outlet obstruction. AB - PURPOSE: There is increasing evidence that ischemia/reperfusion is a major etiological factor in the progression of bladder dysfunction after partial outlet obstruction. If this evidence is correct, treatment with an antioxidant should be beneficial in rabbits subjected to partial outlet obstruction. We designed the current study to determine if diets high in alpha-tocopherol protected the rabbit bladder against dysfunction induced by partial outlet obstruction. MATERIALS AND METHODS: A total of 32 rabbits were separated into 4 groups of 8. Groups 1 and 2 were placed on a diet enriched with 1,000 IU/kg. alpha-tocopherol, and groups 3 and 4 were fed a regular diet containing 44 IU/kg. alpha-tocopherol. After 4 weeks partial outlet obstruction was created in groups 1 and 3, while groups 2 and 4 underwent sham operation. After 4 weeks of obstruction the rabbits were anesthetized and the bladders were rapidly excised. Four longitudinal strips obtained from the bladder body were used for contractility studies. The balance of the bladder body was separated between muscle and mucosa. Each section was frozen and stored at -70C for analysis of malondialdehyde as a measure of peroxidation and for alpha-tocopherol concentrations. RESULTS: Feeding rabbits a diet high in alpha-tocopherol resulted in significant protection against the development of contractile dysfunction after partial outlet obstruction. The protective effect of alpha-tocopherol was related to significantly decreased malondialdehyde and significantly increased tissue concentrations of alpha tocopherol. CONCLUSIONS: These data indicate that a major etiology of bladder dysfunction secondary to partial outlet obstruction is related to free radical generation and resultant membrane lipid peroxidation. PMID- 11435898 TI - Dual effects of ouabain on the regulation of proliferation and apoptosis in human prostatic smooth muscle cells. AB - PURPOSE: To elucidate the role of ouabain in the pathophysiology of benign prostatic hyperplasia we examined the effects of ouabain on the proliferation and apoptosis of human prostatic smooth muscle cells. MATERIALS AND METHODS: Primary cultures of human prostatic smooth muscle cells were obtained from 7 patients with bladder outlet obstruction caused by benign prostatic enlargement. A cell proliferation study was performed using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay method to examine the effects of different concentrations of ouabain and various inhibitors. Western blot analysis was done to determine mitogen activated protein kinase (MAPK) activation. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling reaction method and caspase-3 activity assay were also performed to examine the apoptotic mechanism. RESULTS: Ouabain exhibited a modest but significant proliferative effect in nanomolar concentrations; whereas it induced cell apoptosis at higher concentrations. Ouabain caused rapid activation of p42/44 MAPKs. The proliferative effect of ouabain was completely flattened by W-7 and MAPK kinase (MEK) inhibitor, suggesting the requirement of Ca(2+) mobilization and the involvement of the MEK-p42/44 MAPK cascade. The cytotoxic effect by ouabain was defined as apoptosis and necrosis using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling reaction technique and lactate dehydrogenase release assay, respectively. In addition, ouabain induced profound caspase-3 activity in the cytotoxic concentrations and DEVD-CHO reversed the cytotoxic action to ouabain, demonstrating the involvement of caspase-3 activation in the cytotoxic action. CONCLUSIONS: Ouabain at different concentrations caused dual effects on proliferation and apoptosis in human prostatic smooth muscle cells. At low concentrations ouabain promoted cell proliferation via a Ca(2+) dependent mechanism and activation of the MEK-p42/44 MAPK pathway; whereas it induced cell apoptosis via the activation of caspase-3 activity at higher concentrations. PMID- 11435899 TI - Vascular endothelial growth factor promotes proliferation and migration of cavernous smooth muscle cells. AB - PURPOSE: We investigated whether cavernous smooth muscle cells secrete vascular endothelial growth factor (VEGF), how they respond to VEGF and how age affects their ability to secrete and respond to VEGF. MATERIALS AND METHODS: Corpus cavernous tissues were isolated from rats of various ages and grown as monolayer cell cultures. Smooth muscle identity was determined by immunofluorescence staining. Secreted VEGF was measured with a VEGF enzyme-linked immunosorbent assay kit. Cell proliferation was assayed with a cell titer kit. Cell migration was measured with Transwell chambers. VEGF receptor (VEGFR) messenger RNA and proteins were identified by reverse transcription-polymerase chain reaction and immunoblotting, respectively. RESULTS: Cavernous smooth muscle cells from young (age 1 week) and old (age 28 months) rats secreted the least amounts of VEGF and those from 16-week-old rats secreted the most. VEGF stimulated the growth of cavernous smooth muscle cells at an optimal concentration of 12.5 ng./ml. At this concentration cavernous smooth muscle cells from 11-week-old and 28-month-old rats showed the highest and lowest growth rates, respectively. The optimal VEGF concentration for cavernous smooth muscle cell migration was 10 ng./ml. At this concentration cells from 4-week-old and 28-month-old rats showed the highest and lowest migratory rates, respectively. The expression of VEGFR-1 appeared to parallel the VEGF regulated growth rates of these cells. VEGFR-2 was undetectable in cavernous smooth muscle cells. CONCLUSIONS: Cavernous smooth muscle cells from rats of different ages secreted different amounts of VEGF and showed different abilities to respond to VEGF. Cavernous smooth muscle cells responded to VEGF in terms of cell proliferation and migration. Cavernous smooth muscle cells from rats of different ages expressed different levels of VEGFR-1 but did not express VEGFR-2. PMID- 11435900 TI - Presence of sst2(a) receptor immunoreactivity in rat ependyma and tanycytes. AB - Somatostatin sst2(a) receptor was observed, by immunofluorescence, in ependymal cells and in tanycytes of the wall of the ventricle and the hypothalamic recess of the male rat median eminence. Strong immunoreactivity for the receptor protein was observed in lateral tanycytes (alpha-type) while a moderate signal was seen in medial tanycytes (beta-type). In high magnification the immunoreactive material, of moderate intensity, had a coarse granular appearance. Only few of the alpha-tanycytes also displayed immunoreactive GFAP. The apical portion of the ependymal cells as well as of tanycytes contained immunoreactive S-100 (alphabeta). Since rather high levels of somatostatin are demonstrated to occur in the cerebrospinal fluid of the third ventricle, it is suggested that somatostatin via the sst2(a) receptor may regulate the physiology of tanycytes. PMID- 11435901 TI - Association studies using novel polymorphisms in BACE1 and BACE2. AB - The release of amyloid-beta peptide (Abeta) from beta-amyloid precursor protein (APP) requires cleavage by beta- and gamma-secretases. Several groups have identified a candidate for the beta-site APP-cleaving enzyme, BACE1, and its homologue BACE2. We sequenced the genes for BACE1 and BACE2 and found several polymorphisms in both genes. Genotyping a large cohort of AD cases and controls has shown no association between AD and the intronic polymorphism in BACE2 while there was a weak association between the BACE1 polymorphism in exon 5 and AD in those carrying the APOE epsilon4 allele. PMID- 11435902 TI - Non-undulatory locomotion in the lamprey. AB - The lamprey (a lower vertebrate, cyclostome), in addition to ordinary swimming, is also capable of crawling. Here we describe crawling forward in a narrow U shaped tunnel. A rapid movement along the tunnel was evoked by stimulating the tail. The muscle activity responsible for propulsion was confined to the area around the body bend. Muscles on the inner (concave) side were activated when approaching the turn, and inactivated on the top of the arc. Muscles on the outer (convex) side were co-active with their antagonists, but also active in the area of straightening of the body bend. This pattern of muscle activity propagated along the body. The role of central and reflex mechanisms in the generation of locomotor movements is discussed. PMID- 11435903 TI - Novel cytoplasmic immunolocalization of RNA polymerase II in inclusion-body myositis muscle. AB - Sporadic inclusion-body myositis (IBM) is a progressive degenerative muscle disease of older persons. Abnormalities of gene-expression and RNA metabolism have recently been proposed to contribute to the IBM pathogenic cascade. We now demonstrate, using well characterized, epitope-specific antibodies, that the largest subunit of RNA polymerase II is abnormally accumulated in the cytoplasm of IBM muscle fibers, where it is co-localized with phosphorylated tau on IBM paired helical filaments. Since RNA polymerase II is a crucial nuclear factor involved in both transcription and mRNA processing, our results support the hypothesis that abnormality of either or both of those processes might be caused, in part, by pathological trafficking of RNA polymerase II, and that abnormal trafficking might be an important factor in the IBM pathogenic cascade. PMID- 11435904 TI - Glutamate transporters in the spinal cord of the wobbler mouse. AB - We studied the role of glutamate excitotoxicity in motor neuron degeneration in the wobbler mouse (wr/wr), a model of amyotrophic lateral sclerosis and spinal muscular atrophies. Choline acetyltransferase (ChAT) activity was decreased in the cervical spinal cord and in the muscles innervated by nerves originating in this region of wobbler mice, but no differences were found in the lumbar spinal cord and in the hindleg muscles. Glial fibrillar acid protein (GFAP), a marker of reactive gliosis, was significantly higher in the cervical spinal cord of wobbler mice aged 4 weeks than in controls and the differences were more marked at 12 weeks; no differences were found in the lumbar spinal cord. In spite of this selective degeneration of motor neurons (resulting in strong decrease in the neuronal glutamate transporter EAAC1) and reactive gliosis in the cervical spinal cord, the levels of the glial glutamate transporter proteins GLT-1 and GLAST were similar in wobbler and control mice. Plasma concentrations of excitatory amino acids were no different at any time examined. Our results exclude the involvement of decrease in glutamate GLT 1 transporter in the motor neuron degeneration in wobbler mice. PMID- 11435905 TI - MRI changes in multiple sclerosis following treatment with lofepramine and L phenylalanine. AB - As part of a large, randomized placebo-controlled trial of inpatients with multiple sclerosis (MS), a subsample of 15 underwent cerebral MRI at baseline and 6-months (eight on lofepramine and l-phenylalanine; seven on placebo). Unlike the placebo group, the active group showed a significant reduction in lesion number visible on T1-weighted scans (p < 0.05). The lateral ventricular volume increased, on average, by 1020 mm3 in the untreated group and 600 mm3 in the treated group. In the treated patients the ventricular size change correlated with both change in Gulick MS-related symptoms scale scores (rs = 0.71, p = 0.07) and Gulick MS-related activities of daily living scale scores (rs = -0.83, p = 0.02). It is concluded that treatment with lofepramine and l-phenylalanine is associated with significant MRI changes. PMID- 11435906 TI - Release from masking in fluctuating background noise in a songbird's auditory forebrain. AB - Fluctuations in the ubiquitous masking background noise can be exploited by the vertebrate auditory system to considerably improve signal detection. Here we demonstrate neuronal masking release in amplitude-modulated background noise on the level of the European starling's auditory forebrain, an area that is the analogue of the mammalian primary auditory cortex. Tone-evoked responses in the presence of modulated and unmodulated maskers were recorded in unrestrained birds via radiotelemetry. Based on a rate code, the average amount of neuronal masking release was similar to that observed in a psychoacoustic study on the starling with stimuli confined to a single auditory filter. The results suggest that the neurons exploited predominantly temporal features of the acoustic background to improve signal detection. PMID- 11435907 TI - Adenosine A2A agonist CGS 21680 decreases the affinity of dopamine D2 receptors for dopamine in human striatum. AB - Adenosine A2A receptors (A2AR) and dopamine D2 receptors (D2R) are highly concentrated in the striatum, where they are co-localized and exert reciprocal antagonistic interactions. It has been suggested that the A2R/D2R interactions might provide a therapeutic approach for basal ganglia disorders, such as Parkinson's disease, and schizophrenia. In the present work evidence is presented for the existence of an A2AR/D2R interaction in human brain by using quantitative autoradi- ography. The areas analyzed were the dorsal caudate nucleus and putamen. Parallel studies were performed in rat striatal sections. The A2AR agonist CGS 21680 was found to significantly increase IC50 values of competitive inhibition curves of the D2R/D3R antagonist [125I]iodosulpiride vs dopamine both in rat striatal and human striatal brain sections. PMID- 11435908 TI - White matter lesions and alteration of vascular cell composition in the brain of spontaneously hypertensive rats. AB - There have been few studies on the white matter lesions of spontaneously hypertensive rats (SHR). From the point of view of hypertension and arteriosclerosis, white matter lesions were examined in SHR and stroke-prone SHR (SHRSP), and were then compared with Wistar-Kyoto (WKY) rats. The vasculopathy was analyzed by morphometric immunohistochemistry for collagen and smooth muscle actin. Both SHR and SHRSP had hypertension at > or = 12 weeks of age, and the latter developed severe white matter lesions at 20 weeks. Immuno- histochemistry revealed proliferation of microglia in the white matter and an increase in smooth muscle actin in the vessels of SHRSP compared with the WKY rats and SHR, but there were no changes in the collagen. These results indicate a role of hypertension in the pathogenesis of white matter lesions. However, genetic difference may also be responsible since SHR and SHRSP showed similar hypertension. PMID- 11435909 TI - Heme oxygenase-1 plays an important protective role in experimental autoimmune encephalomyelitis. AB - Increasing evidence shows that oxidative stress plays an important role in the pathogenesis of experimental autoimmune encephalomyelitis (EAE), an animal model of the human disease, multiple sclerosis (MS). Heme oxygenase-1 (HO-1) is a heat shock protein induced by oxidative stress. HO-1 metabolizes heme to the antioxidant bilirubin and carbon monoxide, and represents a powerful endogenous defensive mechanism against free radicals in many diseases. However, the role of this important enzyme in EAE remains unknown. In this study, we showed high expression of HO-1 in lesions of EAE, and demonstrated that hemin, an inducer of HO-1, inhibited EAE effectively. In contrast, tin mesoporphyrin, an inhibitor of HO-1, markedly exacerbated EAE. Our results suggest that endogenous HO-1 plays an important protective role in EAE, and that targeted induction of HO-1 overexpression may represent a new therapy for the treatment of multiple sclerosis. PMID- 11435910 TI - Eye instability induced by vestibular stimulation in rabbits. AB - The slow compensatory phases of the vestibulo-ocular reflex (VOR) in the rabbit tend to drift and the drift reverses the direction. This periodic alternating drift (PAD) has two peculiar characteristics: (1) it is induced by sinusoidal vestibular stimulation in naive animals, being evoked immediately after stimulus onset and persisting after the end of stimulation; (2) the peak velocity and period of the drift are dependent on stimulus amplitude. PAD of the rabbit has strong similarities with PAN, a periodic alternating nystagmus observed in humans with cerbellar disorders and in monkeys after nodulo-uvulectomy, although its peak velocity is smaller. It is hypothesized that PAD is due to a slight instability, caused by vestibular stimulation in darkness, of the cerebellar adaptive loop, which exerts a variable gain control on the time constant of the velocity storage integrator. PMID- 11435911 TI - Error-related brain potentials elicited by vocal errors. AB - Error-related negativity (ERN) is observed immediately after an error in choice reaction time tasks performed by hand response. We examined whether the ERN occurs in relation to slips of vocalization in the Stroop color word task. In one condition, the subject's vocal responses were masked by continuous pink noise in order to prevent vocalization-related cortical potentials from contaminating the ERN time window. This masking procedure was successful in inhibiting the vocalization-related cortical potential. More importantly, vocalization errors elicited a frontocentral negative-going deflection followed by a positive component immediately after the error response regardless of the masking condition. The present results suggest that the error detection mechanism may also elicit an ERN-like component in response to vocal slips. PMID- 11435912 TI - Serial activation of distinct cytoarchitectonic areas of the human S1 cortex after posterior tibial nerve stimulation. AB - MEG recordings visualized non-invasively a dynamic anterior-posterior activation in the pyramidal cell population of the human primary somatosensory cortex (S1) after posterior tibial nerve stimulation. Somatosensory evoked fields (SEFs) were recorded over the foot area in response to right posterior tibial nerve stimulation at the ankle in six normal subjects. A newly developed MEG vector beamformer technique applied to the SEFs revealed two distinct sources in the mesial wall of the left hemisphere around the primary P37m response typically separated by 1.3 cm. The first source was located in area 3b and oriented toward the contralateral hemisphere. The second source was assumed to be in an area near the marginal sulcus and the source orientation was directed posteriorly. The first source began to be active during the initial slope of the P37m. The second source was active after the P37m peak and the signal intensities of the first and second sources were equal at a mean latency of 2.6 ms after the peak of P37m. Then the first source became inactive and the second source was dominant after about 5 ms post-P37m peak. These findings suggest that a single peaked posterior tibial nerve P37m consists of partially overlapping two subcomponents generated in area 3b and an area near the marginal sulcus. PMID- 11435913 TI - Timing and magnitude of frontal activity differentiates refixation and anti saccade performance. AB - EEG data were recorded while 10 subjects generated refixation saccades towards a visual target and antisaccades away from a visual cue. Theoretically, the same basic neural circuitry supports refixation and correct anti-saccade performances, with additional activity in primarily dorsolateral prefrontal cortex circuitry supporting antisaccade-associated inhibitory processes. Analyses demonstrated that sensory registration of visual stimuli is similar for refixation and anti saccade conditions. Increased frontal brain activity at 5 and 15 Hz was observed preceding correct antisaccades when compared to refixation saccades. These analyses provide specific information suggesting that 160-60 ms before saccade generation is the critical period for response inhibition. PMID- 11435914 TI - Flecainide reverses neuropathic pain and suppresses ectopic nerve discharge in rats. AB - We investigated effects of flecainide, a Class IC sodium channel blocker, in the rat chronic constrictive injury (CCI) and ectopic nerve discharge models. In the behavioral evaluation, 2, 6, and 12 mg/kg flecainide were intravenously given to the CCI model, and a dose-dependent analgesic effect was shown on both thermal hyperalgesia and tactile allodynia. In the electrophysiological evaluation using the ectopic nerve discharge model produced by saphenous neurectomy, i.v. administration of 2, 6, and 12 mg/kg flecainide suppressed spontaneous discharge at the peripheral nerve level in a dose-dependent fashion as with the behavioral evaluation, but flecainide did not affect nerve conduction at the dose of 12 mg/kg. PMID- 11435915 TI - Temporomandibular-evoked jaw muscle reflex: role of brain stem NMDA and non-NMDA receptors. AB - This study investigated the possible involvement of brain stem excitatory amino acid receptor mechanisms and the trigeminal subnucleus caudalis (Vc) in temporomandibular joint (TMJ)-evoked reflex jaw muscle activity. Glutamate injected into the TMJ of anesthetized rats reflexly evoked activity in the jaw muscles. Application of lidocaine, but not saline, to the surface of the caudal brainstem overlying Vc significantly suppressed TMJ-evoked jaw muscle activity, while application of NMDA or non-NMDA receptor antagonists also significantly attenuated jaw muscle activity. These results provide evidence that Vc is a critical relay in the TMJ-evoked reflex activation of the jaw muscles, and that both NMDA and non-NMDA receptor mechanisms may contribute to these effects. PMID- 11435916 TI - The cerebellum is involved in predicting the sensory consequences of action. AB - We used H2(15)O PET to examine neural responses to parametrically varied degrees of discrepancy between the predicted and actual sensory consequences of movement. Subjects used their right hand to move a robotic arm. The motion of this robotic arm determined the position of a second foam-tipped robotic arm, which made contact with the subject's left palm. Using this robotic interface, computer controlled delays were introduced between the movement of the right hand and the tactile stimulation on the left. Activity in the right lateral cerebellar cortex showed a positive correlation with delay. These results suggest the cerebellum is involved in signalling the sensory discrepancy between the predicted and actual sensory consequences of movements. PMID- 11435917 TI - Orexin-a activates phospholipase C- and protein kinase C-mediated Ca2+ signaling in dopamine neurons of the ventral tegmental area. AB - The orexin-orexin receptor system has been implicated in the regulation of wakefulness/sleep states. Behavioral and psycho-stimulant effects of orexins have also been shown. Mesolimbic dopamine neurons in the ventral tegmental area (VTA) are implicated in the regulation of reward and wakefulness/sleep, In the present study, we examined the effect of orexin-A on cytosolic [Ca2+]i concentration ([Ca2+]) in the isolated rat VTA dopamine neurons. Orexin-A (10-12-10-8 M) concentration dependently increased [Ca2+]i in dopamine-containing neurons. The [Ca2+]i responses to orexin-A were inhibited under Ca2+-free conditions and by blockers of voltage-gated L- and N-type [Ca2+]i channels, nitrendipine and omega conotoxin, respectively. The [Ca2+]i responses were also abolished by a phosphatidylcholine-specific phospholipase C inhibitor, D609, and a protein kinase C (PKC) inhibitor, calphostin C. A PKC activator, TPA, mimicked orexin-A in increasing [Ca2+]i. These results indicate that orexin-A increases [Ca2+]i in VTA dopamine neurons via phosphatidylcholine-specific PLC- and PKC-mediated activation of L- and N-type Ca2+ channels. This effect may serve as the mechanism by which orexin regulates wakefulness/sleep states and exerts its behavioral and psychostimulant effects. PMID- 11435918 TI - Dynamic activation of distinct cytoarchitectonic areas of the human SI cortex after median nerve stimulation. AB - MEG recordings visualized non-invasively a serial mediolateral activation of the human somatosensory 3b area followed by a stationary activation of area 1 after median nerve stimulation. Somatosensory evoked fields (SEFs) were recorded over the hand area contralateral to the right median nerve stimulation at the wrist in six normal subjects. A newly developed MEG vector beamformer technique applied to the SEFs revealed two distinct sources (areas 3b and 1) in the primary somatosensory cortex (SI) during the primary N20m-P22m response in all subjects. The first source was located in area 3b, which started to move sequentially toward mediolateral direction 0.7 ms prior to the peak of N20m and ended its movement 1.4 ms after the peak with a total distance of 11.2 mm. We speculate that the movement reflects a sequential mediolateral activation of the pyramidal cells in area 3b, which is mediated by horizontal connections running parallel to the cortical surface. The second source in area 1, located 5.6 mm medial and 4.2 mm posterior to the first source, was active 1.0 ms after the N20m peak. Then, the first source became inactive and the second source was dominant. In sharp contrast with the first source, the second source was stationary. The different behavior of these two components (moving vs stationary) indicates independent parallel inputs to area 3b and area 1 from the thalamus. PMID- 11435919 TI - Delayed administration of NGF reverses nerve injury induced central alterations of primary afferents. AB - We have examined whether delayed exogenous NGF administered to an axotomised peripheral nerve reverses the increased transganglionic choleragenoid (CTB) labelling in lamina II. Two, four, eight or 18 weeks after bilateral sciatic nerve section, NGF was applied unilaterally for an additional 2-week period to the transected nerve stump. The transganglionic choleragenoid labelling and substance P (SP) expression were determined and compared to the contralateral axotomised side in the spinal cord dorsal horn. Delayed NGF administration reversed the transganglionic choleragenoid labelling in lamina II when administered 2 or 18 weeks after the sciatic nerve lesion, but not at 4 or 8 weeks. There was also a clear recovery of SP on the axotomised, NGF-treated side 2 or 18 weeks after the sciatic nerve lesion, but not at the intermediate survival times. At the longer survival time, however, there was a recovery of SP regardless of NGF treatment. These results suggest that there is a critical window as to when NGF administration can be effective in reversing axotomy induced changes in the spinal cord. PMID- 11435920 TI - Differential expression of Group I metabotropic glutamate receptors in motoneurons at low and high risk for degeneration in ALS. AB - Glutamate excitotoxicity has been suggested to play a role in amyotrophic lateral sclerosis (ALS), yet it remains unclear why some groups of motoneurons (MNs) are more vulnerable to degeneration than others. Our aim was to compare, in normal adult rats, the expression of Group I metabotropic glutamate receptors (mGluR1 and mGluR5) in MNs normally affected in ALS (XII and spinal MNs) with those which are spared (III and IV MNs). RT-PCR analysis of tissue punches taken from III and XII motor nuclei revealed mRNA for both 'a' and 'b' splice variants of the mGluR1 and mGluR5 receptor subtypes, with expression of the 'a' variant dominant for both receptor subtypes in III and XII nuclei. Immunolabeling for mGluR1a protein was strong in vulnerable (XII and spinal) but negligible in the resistant (III and IV) MNs. Immunoreactivity for mGluR5 was not detected in the cell bodies or proximal dendrites of any MN pool examined. Greater expression of mGluR1a receptor protein within vulnerable MN pools may predispose these neurons to neurodegeneration as seen in ALS. PMID- 11435921 TI - Expression of alpha-synuclein in a human glioma cell line and its up-regulation by interleukin-1beta. AB - Although alpha-synuclein is expressed primarily in neurons, it is a major component of oligodendroglial and astrocytic inclusions in several neurodegenerative diseases. Recent study has further demonstrated that alpha synuclein is expressed in cultured rat oligodendrocytes. We determined whether alpha-synuclein might be expressed in astrocytic cells. alpha-Synuclein mRNA and protein were detected in U251 human astrocytic glioma cells and normal human astrocytes, and the levels were increased in the former, but not in the latter, by stimulation with interleukin-1beta in a time- and concentration-dependent manner. Serum deprivation also led to an increase of alpha-synuclein mRNA and protein in U251 cells. Immunofluorescent staining confirmed the cell-associated alpha-synuclein. These findings suggest that human astrocytes can produce alpha synuclein in culture and that certain inflammatory cytokines and cell stress increase alpha-synuclein expression. PMID- 11435922 TI - Neurotoxic lesions of perirhinal cortex impair visual recognition memory in rhesus monkeys. AB - Recent excitotoxic lesion studies in monkeys have shown that the recognition memory deficits originally attributed to amygdalo-hippocampal damage were due in whole or in part to the accompanying damage to surrounding tissue, including fibers of passage. Here we show that the same conclusion does not apply to the visual recognition impairment produced by aspiration lesions of perirhinal cortex inasmuch as equally severe impairment was found after excitotoxic lesions of this cortex. The finding demonstrates that damage limited to perirhinal neurons is sufficient to impair visual memory and that damage to fibers of passage neither caused nor exacerbated the effect described initially. PMID- 11435923 TI - Estrogen protects against beta-amyloid-induced neurotoxicity in rat hippocampal neurons by activation of Akt. AB - The cellular mechanisms underlying the neuroprotective effects of estrogen are only beginning to be elucidated. Here we examined the role of protein kinase B (Akt) activation in 17beta-estradiol (E2) inhibition of beta-amyloid peptide (31 35) (Abeta31-35)-induced neurotoxicity in cultured rat hippocampal neurons. Abeta31-35 (25-30 betaM) significantly decreased the total number of microtubule associated protein-2 positive cells (MAP2+). This decrease was significantly reversed by pre-treatment with 100 nM E2. Further, 100 nM E2 alone significantly increased the total number of protein kinase B and microtubule associated protein 2 positive cells compared with controls. Such E2-induced increases were inhibited by LY294002 (20 microM), a specific PI3-K inhibitor, as well as by tamoxifen, an estrogen receptor antagonist/selective estrogen receptor modulator. These results indicate that the neuroprotective effects of E2 may be mediated at least in part via estrogen receptor-mediated protein kinase B activation. PMID- 11435924 TI - Hippocampal volume does not change seasonally in a non food-storing songbird. AB - Seasonal differences in hippocampal morphology have been reported in food-storing birds. Non food-storing species have not been investigated however. It is therefore unclear whether seasonal changes in the hippocampus are specifically related to food-storing or reflect a more general seasonal mechanism that occurs in both food-storing and non food-storing birds alike. We determined the volumes of the hippocampal formation and remaining telencephalon in the non-storing male song sparrow (Melospiza melodies morphna) in two experiments comparing birds collected in the spring and fall of 1992-94 (Experiment 1) and 1997 (Experiment 2). Although pronounced seasonal changes in song control nuclei such as the HVC and RA were previously reported for the same brains used in Experiment 1, we found that hippocampal volume did not change with season in either Experiment 1 or 2 for these song sparrow brains. These results suggest that seasonal changes in the hippocampus do not occur in this non food-storing species and may be specific to food-storing birds. PMID- 11435925 TI - Prestin topology: localization of protein epitopes in relation to the plasma membrane. AB - Computer modeling of the outer hair cell (OHC) motor protein prestin produces ambiguous results regarding transmembrane regions and localization of its termini. To determine the location of prestin's N- and C-termini, we created prestin constructs with synthetic epitopes located immediately upstream or downstream of prestin. The spatial distribution of these epitopes was studied in prestin-transfected cells using immunofluorescence. In permeabilized cells, antibodies label the plasma membrane of 30% of the cells, reflecting transfec- tion efficiency. Under non-permeabilizing conditions, the few labeled cells also displayed a lack of plasma membrane integrity. These data suggest that prestin's N-and C-termini are cytoplasmic. Furthermore, prestin staining in OHCs was observed only under permeabilizing conditions. These results implicate prestin's N- and C-termini as portions that may interact with other cytoplasmic proteins. A model of prestin membrane topology is also considered based on the results. PMID- 11435926 TI - Involvement of the sensorimotor cortex in physiological force and action tremor. AB - Whole scalp magnetoencephalography (MEG) signals were recorded in 10 healthy subjects simultaneously with the surface electromyogram (EMG) of the contralateral forearm extensor muscles during isometric contraction and phasic movement of the wrist. In eight subjects, coherence and time domain analyses demonstrated correspondence between the MEG signal, originating near or in the hand region of the motor cortex, and the 6-12 Hz EMG recorded during isometric postural contractions. In contrast, we found little evidence for correspondence between the contralateral EMG and the MEG recorded over the Rolandic region during phasic movements. We conclude that the sensorimotor cortex is differentially involved in physiological force and action tremor at the wrist. PMID- 11435927 TI - Association of immune responses and ischemic brain infarction in rat. AB - Inflammation plays an important role in the pathogenesis of neurodegenerative diseases including ischemia. Occlusion of common carotid artery and middle cerebral artery has been used to produce focal ischemic lesions in the rat. Here, we examined the associations between immune reactions and postischemic brain infarction. Ischemia/reperfusion time-dependently caused brain infarction. The kinetics of inflammatory reactions in rat brain including inflammatory cell infiltration, edema formation, cytokines/chemokines and adhesion molecules production and matrix metalloproteinase activation were relevant to the progression of ischemic infarction. Differential induction profile after ischemia suggests that this activation might contribute to secondary brain damage in ischemic tissues. On the other hand, another possibility of this response is to trigger processes that mediate the neural regeneration after ischemic injury. PMID- 11435928 TI - Binding occurs at early stages of processing in children and adults. AB - Conjoining features in a high-rate serial presentation was studied in children and adults using event-related potentials. Three blocks of trials were run in which targets were defined by colour, by orientation or by a conjunction of colour and orientation. Only adults had faster RTs when detecting targets defined by a single feature than by a combination of features. Compared with adults, children had longer RTs, longer N1 and P2 latencies, and larger P1 and P2 amplitudes. Amplitudes asymmetries were consistent with differing cortical regions being implicated in the processing of colour and form. There were, however, no ERP latency effects as a function of task, suggesting that the binding of features proceeds in parallel with the processing of single features. PMID- 11435929 TI - Expression of estrogen receptor -alpha and -beta immunoreactivity in the cultured neonatal suprachiasmatic nucleus: with special attention to GABAergic neurons. AB - This study investigated the expression patterns of estrogen receptor -alpha (ERalpha) and -beta (ERbeta) in cultured cells of the suprachiasmatic nucleus (SCN) in neonatal rats by combined application of cell culture and double-label immunocytochemistry. The results revealed that the immunoreactivity for either ERalpha or ERbeta (with predominance of ERbeta) was localized in not only neurons but also astrocytes. The co-expression of both ERalpha and ERbeta in the same individual cell was also demonstrated by the double-label immunocytochemistry. The observations also provide a direct evidence for the differential expression of ER subtypes within GABAergic SCN neurons in vitro and suggest that estrogen's effect on the SCN may be mediated at least in part by its ER-containing GABAergic neurons. PMID- 11435930 TI - Convection-enhanced delivery of AAV-2 combined with heparin increases TK gene transfer in the rat brain. AB - Adeno-associated virus type2 (AAV-2) binds to heparan-sulfate proteoglycans on the cell surface. In vivo, attachment of viral particles to cells adjacent to the injection tract limits the distribution of AAV-2 when infused into the CNS parenchyma and heparin co-infusion might decrease the binding of AAV-2 particles to cells in the vicinity of the infusion tract. We have previously shown that heparin co-infusion combined with convection enhanced delivery enhances distribution of the GDNF family trophic factors (heparin-binding proteins) in the rat brain. In this work we show that heparin co-infusion significantly increases the volume of distribution of AAV-2 as demonstrated by immunoreactivity to the transgene product 6 days after infusion into the rat striatum. PMID- 11435931 TI - Estrogen protects neuronal cells from amyloid beta-induced apoptotic cell death. AB - Accumulating studies have shown that estrogen replacement therapy reduces the risk of Alzheimer's disease. In this study, we clarified that 17beta-estradiol (E2) significantly rescues PC12 neuronal cells from amyloid beta protein (Abeta) induced cell death. We found that the amino acid residues of 25 to 35 (Abeta25 35) were more cytotoxic than the full length protein (Abeta1-40) and these residues induced DNA fragmentation typical for apopto- sis. In addition, E2 was confirmed to inhibit calcium influx and cytochrome c release induced by Abeta25 35. Since these sequential events cause apoptosis, the protective effect of E2 may be exerted not by the direct interaction with Abeta, but by the blockade of the mitochondrial apoptotic pathway induced by Abeta. PMID- 11435932 TI - The distribution of BOLD susceptibility effects in the brain is non-Gaussian. AB - A key assumption underlying fMRI analysis in the general linear model is that the underlying distribution of BOLD Susceptibility is gaussian. Analysis of several common data sets and experimental paradigms shows that the underlying distribution for the BOLD signal is non-Gaussian. Further identification shows that the distribution is probably Gamma and implications for hemodynamic modeling are discussed as well as recommendations concerning inferential testing in heavy tailed environments. PMID- 11435933 TI - Increased expression of Fas (CD95/APO-1) in adult rat brain after kainate-induced seizures. AB - Fas (CD95/APO-1), a transmembrane glycoprotein and receptor for the Fas ligand, plays an important role in apoptosis. The present study examined whether excitotoxic cell death induces Fas expression in the adult rat brain. Although relatively light immunostaining was observed in control brain sections, significantly increased Fas immunoreactivity was seen from 4 h to 5 days after the onset of kainic acid-induced seizures. Increased expression of both Fas mRNA and protein were also evident by reverse transcription polymerase chain reaction and Western blotting, respectively. Fas induction was correlated with neuronal apoptosis as demonstrated by colocalization of Fas and terminal dT-mediated dUTP nick end-labeling (TUNEL). Cells with increased Fas-expression were also immunoreactive for tumor suppressor p53 and neuronal specific nuclear protein (NeuN). These results suggest that Fas receptor may contribute to excitotoxic neuronal death in cooperation with p53, and further implicates the Fas pathway in the pathophysiology of neurodegenerative diseases. PMID- 11435934 TI - Hypothermia reduces glutamate efflux in perilymph following transient cochlear ischemia. AB - The effect of hypothermia on ischemic injury of the cochlea in gerbils was studied with particular regard to glutamate efflux in the perilymph. Under normothermic conditions interruption of the blood supply to the cochlea for 15 min caused a remarkable elevation of the compound action potential (CAP) threshold, and an increase in perilymphatic glutamate. The CAP threshold recovered to some extent with reperfusion, but not to preischemic levels. CAP thresholds, under hypothermic conditions and with reperfusion, recovered promptly to near pre-ischemic levels, while glutamate concentration did not change. These results, together with electron microscopy studies, suggest that hypothermia prevents hearing loss primarily through reduction of glutamate efflux at the synopses between inner hair cells and primary afferent auditory neurons. PMID- 11435935 TI - Decrease of adenylyl cyclase activity and expression by a sigma1 receptor ligand and putative atypical antipsychotic. AB - We examined whether changes in the adenylyl cyclase system could be induced by the administration of the sigma1 receptor ligand and putative atypical antipsychotic 4-[4-fluorophenyl]-1,2,3,6-tetrahydro-1-[4-[1,-2,4-triazol-1 il]butyl]pyridine citrate) (E-5842). Repeated (21 days) but not acute (2 h) treatment with E-5842 induced a significant decrease in adenylyl cyclase type I immunoreactivity and adenylyl cyclase activity in rat frontal cortex membranes, with less or no effect in other brain regions such as the hippocampus or the striatum. Changes in immunoreactivity were not observed in other adenylyl cyclases (type V/VI). The reported changes, observed only after a chronic treatment, could be related to the mechanism of action of sigma receptor ligands in general or to that of E-5842 in particular and should be taken into account, given the long duration of treatment in psychiatric patients. PMID- 11435936 TI - Event-related brain potentials in normal children during detection of inverse serial digits. AB - Three inverse serial digit detection tasks were evaluated with event-related brain potentials (ERPs) in 15 11-year-old children to determine how the increase of perceptual or memory demands could modify detection processing. Reaction times were significantly longer for the task that used visual blurring, compared to that with a greater memory demand. Difference-ERPs (target minus non-target conditions) showed three significant parietal components; one earlier positive peak at 162 ms interpreted as an index of working memory load; a same polarity 295 ms peak which probably represents a P3 analogous and a subsequent negative polarity component (520 ms) possibly involved with motor preparation. A fourth difference-component was a frontal positive peak at 680 ms, interpreted as related to task difficulty. PMID- 11435937 TI - Metabolic and electrophysiological alterations in an animal model of neocortical neuronal migration disorder. AB - Cortical migration disorders are a major cause for intractable epilepsy syndromes. High resolution MRI and PET are increasingly capable to identify cortical dysgenesis. In this study we used the rat freeze lesion model to investigate cortical morphological and functional changes in adult rats after induction of a cortical freeze lesion at postnatal day (p) 0. Autoradiographic measurements of basic cortical [14C]deoxyglucose metabolism showed a significant reduction up to 1 mm lateral to the lesion but no remote changes. Electrophysiological in vitro recordings revealed a significant reduction in the amplitude of stimulus-evoked field potential responses recorded lateral to the lesion as compared to medial recording sites. Our data provide further evidence that spatially restricted developmental alterations of cortical morphology cause functional changes in surrounding and histologically normal areas that need to be considered for a better understanding of the resulting pathophysiology. PMID- 11435938 TI - Compartmentalized tau hyperphosphorylation and increased levels of kinases in transgenic mice. AB - The formation of neurofibrillary tangles in Alzheimer's disease is preceded by a pretangle stage of hyperphosphorylated tau. To characterize pretangle tau in vivo, we correlated, in human tau transgenic mice, levels of kinases known to phosphorylate tau in vitro with the phosphorylation of tau at specific epitopes. Levels of cyclin-dependent kinase-5 were increased in axons of CA1 pyramidal neurons, where tau was phosphorylated specifically at the AD2 epitope Ser396/Ser404. The 12E8 epitope serine262/serine356 and the AT180 epitope threonine231/serine235 were phosphorylated in dendrites, and colocalized with increased levels of glycogen synthase kinase-3. CA1 neurons phosphorylated tau at more epitopes than dentate gyrus neurons, suggesting that tau phosphorylation is cell type-specific, a possible explanation for the spatial distribution of neurofibrillary tangles. PMID- 11435939 TI - Orexin receptors couple to Ca2+ channels different from store-operated Ca2+ channels. AB - We have investigated Ca2+ release and receptor- and store-operated Ca2+ influxes in Chinese hamster ovary-K1 cells expressing human OX1 orexin receptor. Receptor operated Ca2+ influx-response to 3 nM orexin-A was not affected by Gd3+ or 2-APB (2-aminoethoxydiphenyl borate), but was inhibited by Ni2+. Store-operated Ca2+ influx was blocked by Ni2+, Gd3+ and 2-APB, whereas the thapsigargin-induced release was unaffected. 2-APB did not block inositol-1,4,5- trisphosphate dependent Ca2+ release in these cells. Thus, low concentrations of orexin-A cause activation of two Ca2+ influxes in the cells: primarily, a receptor-operated Ca2+ influx, and secondarily, a store-depletion activated Ca2+ influx, which is subsequent to receptor-activated Ca2+ influx and the therewith-caused IP3 production. The results show that these two rely on different molecular entities. PMID- 11435940 TI - Counter-stimulatory effects on pain perception and processing are significantly altered by attention: an fMRI study. AB - Counter-stimulation reduces pain perception; however, the role of attention during this process is rarely discussed despite attention itself being a well known modulator of pain perception. This study investigated the effect of attentional modulation on pain perception during counter-stimulation using fMRI. Subjects received a noxious thermal stimulus together with an innocuous vibratory counter-stimulus. Subjects directed their attention towards either pain, vibration, or a neutral visual stimulus. During painful and counter-stimulation all subjects reported a reduction in pain perception when attending to counter stimulation compared with attending to pain. Imaging data supported this behavioural finding showing reduced activity in pain processing areas (anterior cingulate, insula, thalamus). These results suggest attention plays an important part in the pain relief experienced from counter-stimulation. PMID- 11435941 TI - Brain activation during odor perception in males and females. AB - Several studies indicate that women outperform men in olfactory identification tasks. The psychophysical data are more divergent when it comes to gender differences at levels of odor processing which are cognitively less demanding. We therefore compared cerebral activation with H2(15)O PET in 12 females and 11 males during birhinal passive smelling of odors and odorless air. The odorous compounds (odorants) were pure olfactory, or mixed olfactory and weakly trigeminal. Using odorless air as the baseline condition, activations were found bilaterally in the amygdala, piriform and insular cortices in both sexes, irrespective of the odor. No gender difference was detected in the pattern of cerebral activation (random effect analysis SPM99, corrected p < 0.05) or in the subjective perception of odors. Males and females seem to use similar cerebral circuits during the passive perception of odors. The reported female superiority in assessing olfactory information including odor identification is probably an effect of a difference at a cognitive, rather than perceptive level of olfactory processing. PMID- 11435942 TI - Glutamate inhibition of NMDA-induced hydroxyl radical release: an ontogenic study in rat. AB - Hydroxyl radicals (.OH) are frequently associated with glutamate excitotoxicity and may be critical in the occurrence of perinatal brain damage. We thus investigated the mechanisms regulating the glutamate-induced release of toxic.OH during development, using microdialysis and salicylate as an.OH trap. Glutamate inhibited.OH release until post-natal day 14, but stimulated this release from day 21 onwards. DHPG [(RS)-3,5-dihydroxyphenylglycine], a group-I metabotropic glutamate receptor agonist, similarly reduced the.OH release at day 14, but was ineffective afterwards. DHPG also completely blunted the tremendous NMDA induced.OH release at day 14 but not at day 21. Glutamate itself therefore tonically inhibited a possible free radical release through NMDA channel activation during early development. PMID- 11435943 TI - The thalamus interrupts top-down attentional control for permitting exploratory shiftings to sensory signals. AB - When attention is involuntarily drawn in a direction different to that of the target, slower motor response times are observed (i.e. the meridian effect). Previous data suggested that the thalamus might participate in the generation of visual salience. What may be the role of the thalamus in the capture by luminance transients when attentional control is in action? A single experiment was administrated in a group of ten healthy volunteers as well as in a group of three patients with unilateral thalamic infarcts. Subjects participated in a task where attentional control was interrupted by a distractor. The meridian effect was present only in the performance of the healthy volunteers and when distractors occurred in the ipsilesional (intact) hemifield of the thalamic patients. These results suggest that when an important signal appears during attentional focalization, the thalamus interrupts current focalization and permits the compilation of an attentional program in the midbrain aiming at generating an orienting response towards the source of this signal. PMID- 11435944 TI - GAPDH knockdown rescues mesencephalic dopaminergic neurons from MPP+ -induced apoptosis. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12) has a number of diverse functions apart from glycolytic function. We explored the possible involvement of GAPDH in 1-methyl-4-phenylpyridinium (MPP+)-induced death of mesencephalic dopaminergic neurons (MDNs) in culture. MPP+ (10 and 20 microM, 24 h) exposure selectively decreased the survival of tyrosine hydroxylase positive (TH+) MDNs, which manifested apoptotic features including shrinkage of the cell body, chromatin condensation and nuclear fragmentation. Two types of GAPDH antisense oligonucleotides almost completely rescued MDNs from MPP+ toxicity. GAPDH was strongly expressed in apoptotic TH+ neurons, and MPP+ exposure significantly increased the percentage of TH+ neurons in which GAPDH is over expressed. Confocal microscopic analysis demonstrated the nuclear accumulation of GAPDH in neurons undergoing MPP+-induced apoptosis. These results suggest that MPP+ causes apoptosis of MDNs, concomitant with the over-expression and nuclear accumulation of GAPDH. PMID- 11435945 TI - Burrowing into prion disease. AB - Mice received intra-hippocampal injections of scrapie-infected brain homogenate. Open field activity increased from around week 12 post-injection. Concomitantly the tendency to displace food from a tube inside the home cage decreased. The food was generally dug out with the feet, rather than carried by mouth, so its displacement was called burrowing. Food restriction was unnecessary for this burrowing to occur. Only later, around 18 weeks, did more general motor impairments develop. As burrowing in scrapie-infected mice decreased when open field activity increased, and preceded later motor impairments, it was not due to motor dysfunction. Burrowing is a simple, sensitive, objective, ethological measure, sensitive to preclinical prion disease. Other potential applications are in transgenic and knockout mice, models of ageing and Alzheimer's disease, and pharmacology, particularly neuroleptics. PMID- 11435946 TI - Regulation of chemokines and chemokine receptors after experimental closed head injury. AB - The expression of the chemokines macrophage inflammatory protein (MIP)-2 and MIP 1alpha and of their receptors CXCR2 and CCR5 was assessed in wild type (WT) and TNF/lymphotoxin-alpha knockout (TNF/LT-alpha-/-) mice subjected to closed head injury (CHI). At 4 h after trauma intracerebral MIP-2 and MIP-1alpha levels were increased in both groups with MIP-2 concentrations being significantly higher in WT than in TNF/LT-alpha-/- animals (p < 0.05). Thereafter, MIP-2 production declined rapidly, whereas MIP-1alpha remained elevated for 7 days. Expression of CXCR2 was confined to astrocytes and increased dramatically within 24 h in both mouse types. Contrarily, CCR5 expression remained constitutively low and was mainly localized to microglia. These results show that after CHI, chemokines and their receptors are regulated differentially and with independent kinetics. PMID- 11435947 TI - General and task-specific frontal lobe recruitment in older adults during executive processes: a fMRI investigation of task-switching. AB - Performance deteriorates when subjects must shift between two different tasks relative to performing either task separately. This switching cost is thought to result from executive processes that are not inherent to the component operations of either task when performed alone. Medial and dorsolateral frontal cortices are theorized to subserve these executive processes. Here we show that larger areas of activation were seen in dorsolateral and medial frontal cortex in both younger and older adults during switching than repeating conditions, confirming the role of these frontal brain regions in executive processes. Younger subjects activated these medial and dorsolateral frontal cortices only when switching between tasks; in contrast, older subjects recruited similar frontal regions while performing the tasks in isolation as well as alternating between them. Older adults recruit medial and dorsolateral frontal areas, and the processes computed by these areas, even when no such demands are intrinsic to the current task conditions. This neural recruitment may be useful in offsetting the declines in cognitive function associated with ageing. PMID- 11435948 TI - The Rb pathway in neurogenesis. AB - Cell division during embryogenesis plays a crucial role in the formation of the nervous system. During this developmental process, proliferating neural precursor cells commit to a neuronal fate and, as a consequence, undergo terminal mitosis and adopt a neuronal phenotype. A key cell cycle regulator, the tumor suppressor protein, retinoblastoma (Rb), is involved in both terminal mitosis and neuronal differentiation. Neural development is a complex process involving cell proliferation, cell fate determination and differentiation, as well as programmed cell death. In this review, we will examine each of these processes in turn, focussing on the role of the Rb family proteins to examine their many influences on these events. PMID- 11435949 TI - The hen house. PMID- 11435950 TI - Focus on primary care: pruritic dermatological conditions in pregnancy. AB - This article reviews the literature that evaluates pruritic urticarial papules and plaques of pregnancy, herpes gestationis, and intrahepatic cholestasis of pregnancy and their impact on the fetus. Using MEDLINE years 1966 to 1999, a literature search was performed using the terms pregnancy, dermatology, pruritic urticarial papules and plaques of pregnancy, herpes gestationis, and intrahepatic cholestasis of pregnancy. References from the selected papers were then reviewed for additional sources. Thirty-seven studies were reviewed. Both original studies and review articles were included in the sources. The results of each study as originally reported are included to provide the reader the finding of each. The available literature reports no risk with pruritic urticarial papules and plaques of pregnancy; however, the current opinion of most was that there is an increased risk with herpes gestationis and intrahepatic papules and plaques of pregnancy. Although much information is known concerning these unique conditions, a consensus regarding their effect on the fetus has yet to be reached. Pregnancies affected by herpes gestationis and cholestasis of pregnancy should be considered high risk until more definitive evidence can be gained. PMID- 11435951 TI - Cerebral palsy and chorioamnionitis: the inflammatory cytokine link. AB - Cerebral palsy remains a significant cause of perinatal morbidity in medically developed countries. Human epidemiologic data suggest a relationship between cerebral palsy and chorioamnionitis mediated by proinflammatory cytokines. This association has been confirmed by experimental data from human and animal research that demonstrate an increase in cytokine levels in the amniotic fluid of cases of white matter damage. Recent evidence suggests this damage is the result of a fetal inflammatory response initiated in response to placental inflammation. The strong association between cerebral palsy and chorioamnionitis warrants additional investigation into the mechanisms by which white matter damage is initiated and into possible neuroprotective treatments to prevent the development of cerebral palsy. PMID- 11435952 TI - Implications of the Human Genome Project for obstetrics and gynecology. AB - The initial sequencing of the human genome should be regarded as a milestone in a road that stretches years into the future; the full ramifications of the Human Genome Project are still only being theorized. Researchers will benefit from the catalog of human genes in studies of the genetics of disease susceptibility and the cell biology of gene interactions. Clinicians will increasingly offer genetic or biochemical testing to identify those at highest risk for a number of diseases. Drug discovery will eventually follow newly possible studies of gene expression and protein function. However the Human Genome Project eventually shapes medicine, it is certain that physicians, particularly obstetricians and gynecologists, will need to be well versed in the scientific and ethical issues involved, inasmuch as we will likely be at the center of the most heated debates. PMID- 11435953 TI - Heat shock proteins, anti-heat shock protein reactivity and allograft rejection. AB - Heat shock proteins are families of highly conserved immunodominant molecules, reactivity to which has been implicated in the pathogenesis of a number of autoimmune and vascular disease states. However, heat shock proteins are cytoprotective, and in clinical and experimental arthritis, anti-heat shock protein reactivity can down modulate immune responses via a self-Hsp reactive, Th2-type mechanism. Despite a number of studies associating heat shock protein expression and anti-heat shock protein reactivity with allograft rejection, the balance between protective and damaging effects and the precise influence of these responses on graft outcome is unclear. This article reviews current knowledge surrounding heat shock proteins, autoimmunity, and allograft rejection and presents a perspective on the potential influence of these proteins and the stress response on allograft outcome. PMID- 11435954 TI - Hemodynamic monitoring during xenotransplantation. PMID- 11435955 TI - Re: a case-control comparison of the results of renal transplantation from heart beating and non-heart-beating donors. PMID- 11435956 TI - Does the immunogenicity of cadaver donor kidneys influence early posttransplant events? PMID- 11435957 TI - Hemodynamic consequences of porcine kidney xenograft reperfusion in cynomolgus monkeys. AB - BACKGROUND: After xenograft reperfusion, complement activation may lead to generation of anaphylatoxins and cardiocirculatory instability of the recipient. METHODS: In 13 cynomolgus recipients of either unmodified or human decay accelerating factor transgenic porcine kidneys cardiocirculatory parameters were measured by single indicator transpulmonary thermodilution. RESULTS: After graft reperfusion, recipient cardiac output decreased by 25.4% (P<0.05), intrathoracic blood volume by 22.8% (P<0.05), extravascular lung water increased slightly (P=n.s.). The impairment in cardiac output was neither influenced by the graft's weight or human decay accelerating factor transgenicity. sC3a and sC5b-9 complement levels in the recipient monkeys showed a sharp peak upon reperfusion. CONCLUSIONS: After reperfusion a marked and significant cardiodepression accompanied by relative volume depletion were observed. Analysis of volume status ruled out a mere volume shift as the underlying reason for the observed drop in cardiac output. These data may be relevant for the perioperative management of human recipients of discordant xenografts in the future. PMID- 11435958 TI - Deuterium nuclear magnetic resonance for evaluating the metabolic status of livers subjected to warm ischemia. AB - BACKGROUND: The development of reliable methods for assessing the viability of currently available livers is expected to increase the number of successful transplantations. METHODS: 2 H nuclear magnetic resonance (NMR) was used to search for metabolic markers of ischemia in explanted rat livers. Deuterium oxide (2 H2O) was used as a source of 2 H. A total of 10-80% v/v 2 H2O was added to homogenates obtained from a liver biopsy and the formation of 2 H-labeled metabolites was monitored. RESULTS: Some well-resolved 2 H resonances were found in the homogenates from biopsies of warm ischemic liver. Two of these were identified as [3-2 H] lactate and [2-2 H] lactate, and a linear relationship was found between the ratio of [[2-2 H] lactate] to [[3-2 H] lactate] and the warm ischemia time. The deuterium incorporation into lactate was explained on the basis of the metabolic events occurring under hypoxic conditions. CONCLUSIONS: The experimental results support the application of 2 H NMR for a reliable evaluation of the metabolic status of a liver harvested from non-heart-beating donors. PMID- 11435959 TI - Porcine neonatal pancreatic cell clusters in tissue culture: benefits of serum and immobilization in alginate hydrogel. AB - Porcine neonatal pancreatic cell clusters (NPCCs) may be a suitable source of insulin producing tissue for transplantation in diabetic patients. The possible beneficial effect of serum on maturation of NPCCs in vitro is difficult to achieve because of cell clumping, which can be avoided by immobilization in alginate hydrogel matrix. Collagenase treated pancreata, cultured for 4 days, formed NPCCs that were embedded in alginate cross-linked with CaCl2 and cultured in modified Ham's F10 medium with 10% fetal calf serum (FCS) for 10 days. NPCCs cultured as suspension in F10+ with 0.5% bovine serum albumin or with 10% FCS were used as control. To prevent the aggregation when cultured with serum, NPCCs were kept as a very diluted suspension. At the beginning and end of the culture, samples were taken for insulin and DNA content and immunostained for beta and non beta cells. The culture of NPCCs immobilized in alginate resulted with 3-fold increase in insulin content and 9-fold increase in insulin/DNA ratio. Histology revealed evident increase of number of insulin- and other hormone-positive cells compared with the control. Even though 2 weeks in culture resulted in impaired glucose-induced insulin release, the amount of insulin secreted by clusters cultured in the presence of serum was 4-fold higher than in serum-free conditions. After transplantation, NPCCs retrieved from alginate reversed hyperglycemia similarly to NPCCs cultured in standard conditions. In conclusion, this study shows the feasibility of in vitro immobilization of NPCCs in alginate three-dimensional matrix, allowing cell clusters to be cultured at least two times higher density compared with culture in suspension. PMID- 11435960 TI - Photodynamic therapy with motexafin lutetium (Lu-Tex) reduces experimental graft coronary artery disease. AB - BACKGROUND: Motexafin lutetium (Lu-Tex) is a photodynamic therapy (PDT) agent that localizes in atheromatous plaque in which it can be activated by far-red light. Lu-Tex biolocalization was examined in graft coronary artery disease (GCAD) with a rodent allograft model. After photoactivation, the effect on intimal proliferation was assessed. METHODS: A PVG to ACI rat heterotopic heart transplantation model was used. Lu-Tex (10 mg/kg) was intravenously administered 90 days after transplantation. Photoactivation was performed 24 hr after Lu-Tex administration. A light-emitting diode, central wavelength of 742 nm, was used to illuminate the intraperitoneally placed allografts via a laparotomy (light fluence of 75 J/cm2 at a power density of 75 mW/cm2). Animals were divided into four groups according to postoperative treatments: PDT with Lu-Tex injection and light illumination (n=21), Lu-Tex injection and laparotomy (n=14), laparotomy with light only (n=14), and laparotomy only (n=16). GCAD was quantitatively assessed 14 days after treatments. RESULTS: Lu-Tex localized in atherosclerotic plaque in vessels with GCAD. PDT significantly reduced both the percent of affected vessels and intimal proliferation compared to all other control study groups. alpha-Smooth muscle cell actin and anti-rat macrophage antibody-positive areas were significantly reduced within the neointima in allografts treated with PDT compared to all other study groups. CONCLUSIONS: PDT significantly reduced atherosclerotic lesions of GCAD. Lu-Tex-mediated PDT may, therefore, be a potential method for treating accelerated atherosclerosis associated with transplantation. PMID- 11435961 TI - T cell and B cell tolerance to GALalpha1,3GAL-expressing heart xenografts is achieved in alpha1,3-galactosyltransferase-deficient mice by nonmyeloablative induction of mixed chimerism. AB - BACKGROUND: We have previously demonstrated that mixed xenogeneic chimerism and donor-specific T-cell tolerance can be induced in the rat-to-mouse species combination by using a relatively nontoxic, nonmyeloablative conditioning regimen. However, natural antibodies (NAbs) against Galalpha1,3Gal (Gal) pose an additional major barrier to pig-to-human vascularized xenograft acceptance. METHODS: To determine whether the mixed chimerism approach could also overcome this humoral barrier, T cell-depleted rat (GalT+/+) bone marrow cells (BMC) were transplanted to alpha1,3-galactosyltransferase deficient (GalT-/-) mice conditioned with a nonmyeloablative regimen, consisting of transient T cell and natural killer (NK) cell depletion, 3 Gy whole body irradiation, and 7 Gy thymic irradiation. RESULTS: By giving a high dose (180x106) of rat BMC, persistent mixed chimerism could be induced in GalT-/- mice, although the level of donor type hematopoietic repopulation declined over time. Induction of mixed chimerism was associated with a rapid disappearance of anti-Gal and anti-rat NAb in the sera. Both anti-Gal Ab-producing cells and B cells with receptors recognizing Gal were undetectable in mixed chimeras, even when the chimerism levels declined, suggesting that a very low level of chimerism could effectively maintain B-cell tolerance to Gal, probably by clonal deletion and/or receptor editing. Mixed chimeras accepted subsequently transplanted donor-type rat hearts (>100 days) without immunosuppressive therapy, whereas delayed vascular and even hyperacute rejection of rat hearts occurred in conditioned control GalT-/- mice. Cellular rejection occurred by 5-6 days in conditioned control wild-type mice. CONCLUSIONS: These findings demonstrate that induction of mixed chimerism with a nonmyeloablative regimen can prevent vascularized xenograft rejection by cellular and anti-Gal Ab-dependent pathways in GalT+/+-to-GalT-/- species combinations. PMID- 11435962 TI - L-arginine polymer mediated inhibition of graft coronary artery disease after cardiac transplantation. AB - BACKGROUND: Nitric oxide (NO) limits the development of graft coronary artery disease (GCAD) in transplanted hearts. We hypothesized that l-arginine polymers administered to cardiac allografts ex vivo would translocate across vascular cellular membranes, up-regulate inducible nitric oxide synthase (iNOS) production of NO, and inhibit the development of GCAD. METHODS: Three groups of PVG rat donor hearts were incubated with either 0.8 ml phosphate-buffered saline, (PBS, n=12) or 50 microM L-arginine polymer solutions of length five (R5, n=12) or nine (R9, n=12) prior to heterotopic transplantation into ACI recipients. Graft vessels were scored at POD 60 and 90 for percentage luminal narrowing (%LN), intima to media ratio (I/M), and percentage affected vessels (%AV). Translocation efficiency was determined by treatment with biotinylated polymers. NO production of treated aortic segments was determined in vitro by Griess reaction. RESULTS: Translocation efficiencies were 89+/-19% (R9), 7+/-10% (R5), and 0+/-0% PBS (ANOVA, P<0.001) which corresponded to NO production in treated aortic segments of 0.175+/-0.17 (R9), 0.120+/-0.006 (R5), and 0.135+/-0.035 microM/mg (PBS), (ANOVA, P=0.002). GCAD scores at POD 60 were: %LN: 3.2+/-3.8% (R9), 12.6+/-6.7% (R5), 11.3+/-4.2% (PBS) (ANOVA, P=0.025); I/M: 0.03+/-0.04 (R9), 0.13+/-0.07 (R5), 0.12+/-0.05 (PBS) (ANOVA, P=0.037); %AV: 7+/-7% (R9), 19+/-7%(R5), 22+/ 9%(PBS) (ANOVA, P=0.021). Reduction of GCAD parameters was maintained at POD 90. CONCLUSION: R9 efficiently translocated across cytoplasmic membranes, enhanced vascular NO production, and decreased neointimal hyperplasia. This ex vivo treatment may have a therapeutic role in preventing GCAD. PMID- 11435963 TI - alpha4 integrin in islet allograft rejection. AB - BACKGROUND: Adhesion molecules are involved in multiple steps of the continuum of allograft rejection. We studied the effects of blockade of the interactions between alpha4 integrin and its ligands, vascular cell adhesion molecule-1 (VCAM 1) and fibronectin, on allograft survival. METHODS: Streptozotocin-induced diabetic CBA (H-2k) mice received islet transplants from BALB/c (H-2d) donors. Recipient mice were treated with antibodies against alpha4 integrin (PS/2), VCAM 1 (MK 2.7), and a peptide corresponding to the binding site of alpha4 integrin on fibronectin (connecting segment 1 peptide, CS1-peptide). Graft function was measured by daily tail vein blood glucose levels, with rejection defined as the return of hyperglycemia. Graft-bearing kidneys were removed for immunohistochemical analysis. RESULTS: Treatment with anti-alpha4 integrin antibody, anti-VCAM-1 antibody, or with CS1-peptide led to long-term survival of islet allografts. Recipients with long-surviving islet grafts did not show tolerance, in that they rejected a second donor-type islet allograft. Although both anti-alpha4 integrin antibody and CS1-peptide completely abolished cellular infiltration of the islet graft 7 days after transplantation, anti-VCAM-1-treated recipients showed a dense peri-islet infiltrate of activated, alpha4 integrin+, cytotoxic T cells. CONCLUSIONS: These data show that alpha4 integrin is critically important to allograft rejection. Anti-VCAM-1 antibody appears to prevent rejection without qualitatively affecting either T cell activation or migration to the graft. Conversely, anti-alpha4 integrin antibody and CS1-peptide may prevent islet allograft rejection by altering either T cell activation or lymphocyte trafficking. Blocking interactions between alpha4 integrin and its ligands may provide novel forms of immunosuppression. PMID- 11435964 TI - A case-control comparison of the results of renal transplantation from heart beating and non-heart-beating donors. AB - INTRODUCTION: The decline in heart-beating brainstem dead organ donors has necessitated the search for other organ sources. In the field of renal transplantation one alternative source currently available, but little used, is that of kidneys from non-heart-beating donors (NHBD). Reticence to use NHBD kidneys is in part due to concerns over the effect that warm ischemic may have subsequent graft function. Presented here are the results of the NHBD renal transplants at the Leicester transplant unit, and compared with matched heart beating donor transplants as a case control analysis. METHODS: In order to analyze any differences in graft performance between the two organ sources, the confounding effect of other variables known to influence the outcome of renal transplantation was minimized by matching NHBD and HBD transplants for the following criteria: donor age and sex, first or re-transplant, anastomosis and cold times, tissue match and PRA sensitisation. Transplant performance was assessed primarily by graft survival, the statistical evaluation of which was by log rank analysis of Kaplan-Meier curves. RESULTS: 72 NHBD and 192 HBD transplants were performed over an eight year period. Of the 192 HBD transplants, 105 matched one or more of the NHBD by the criteria outlined above, and thus constituted the control group for comparison. There was no significant difference in overall graft survival between the two groups. The 5 year survival for the NHBD was 73% compared with 65% for HBD kidneys. When death with a functioning graft is treated as censored data, then these figures become 75% and 81% respectively, again without statistical significance. CONCLUSION: NHBD kidneys are a valuable additional source of organs for transplantation, with long-term survival, comparable to transplants from HBD. PMID- 11435965 TI - Prediction of the long-term metabolic success of the pancreatic graft function. AB - BACKGROUND: Strategies to prevent the return to the diabetic state for graft loss or failure or any other cause after pancreas transplantation require the identification of the subjects at risk. This study evaluated whether daily glucose, insulin, and c-peptide profiles and studies of insulin sensitivity and secretion after transplantation predict pancreatic graft failure. METHODS: Fifty three subjects with type 1 diabetes with end-stage renal failure who received a combined pancreas and kidney transplant underwent the following procedures 1 year after transplantation: 1-day metabolic profiles, sampling every 2 hours for plasma glucose, serum insulin, and c-peptide (n=51); an intravenous glucose tolerance test (IVGTT) to evaluate insulin secretion (n=48); and an euglycemic insulin clamp to evaluate insulin sensitivity (M value, n=14). The recipients were then followed up to 8 years (mean follow-up 4.8+/-0.3 years) to evaluate the return to the diabetic state. RESULTS: Survival analysis showed that plasma glucose in the profiles and insulin secretion in IVGTT were strongly related to the risk of returning to the diabetic state. A cutoff value of mean daily plasma glucose >127 mg/dL, corresponding to the top quartile of the mean plasma glucose distribution in the profiles, predicted the return to the diabetic state within 4 years from transplantation with a 93% specificity and a 100% sensitivity. A cutoff value of insulin delta peak <32 microU/ml in the IVGTT predicted the return to the diabetic state within 4 years from transplantation with a 75% specificity and a 75% sensitivity. In contrast, the M value in the clamp was devoid of predictive value. CONCLUSIONS: This study indicates that the mean 24-h plasma glucose 1 year after transplantation is the strongest predictor of the return to the diabetic state. The risk is related to defects in insulin secretion and not to insulin resistance. Metabolic profiles can be used to screen the subjects at risk to strictly monitor the graft function and to investigate early determinants of graft failure. PMID- 11435966 TI - Are parenchymal changes in early post-transplant biopsies related to preservation reperfusion injury or rejection? AB - BACKGROUND: The progression of parenchymal changes in liver allograft biopsies due to preservation-reperfusion injury (PRI) and their differentiation from rejection related changes is poorly understood. The aim of this study was to determine which changes in a 1-week posttransplant biopsy could be attributed to PRI and which to acute rejection. METHODS: One week protocol liver transplant biopsies from patients with mild PRI (day 1 AST<400 IU/L) were compared with those from patients with severe PRI (day 1 AST>2000 IU/L). Parenchymal changes (cholestasis, ballooning, steatosis, necrosis) and rejection-related inflammatory features (portal tract inflammation, bile duct inflammation, portal vein endothelial inflammation, hepatic vein endothelial inflammation, and centrilobular inflammation) were blindly assessed semiquantitatively. RESULTS: Fat, cholestasis, and hepatocyte ballooning were significantly worse in the severe PRI group, and these features showed no correlation with histological features related to acute rejection. Centrilobular hepatocyte necrosis correlated with hepatic venular endothelial inflammation and centrilobular inflammation but not with rejection related features in portal tracts or with PRI. These findings suggest that centrilobular necrosis is a manifestation of a rejection-related parenchymal injury and may involve different pathogenetic mechanisms to rejection related features in portal tracts. CONCLUSIONS: This study indicates that in early posttransplant biopsies, fat, cholestasis, and ballooning can largely be attributed to PRI. By contrast, centrilobular hepatocyte loss should be suspected as a rejection related phenomenon, even if typical portal tract changes are not prominent, and augmentation of immunosuppression should be considered. PMID- 11435967 TI - Economic analysis of basiliximab in renal transplantation. AB - BACKGROUND: Basiliximab is a chimeric monoclonal directed against the alpha-chain of the interleukin-2 receptor. International studies have shown that it is highly effective in preventing acute rejection in patients receiving Neoral, and causes no measurable incremental toxicity, but its economic value remains unknown. METHODS: This study employed an economic model to examine the potential economic benefit of basiliximab. Parameter estimates were derived from a randomized, prospective, double-blind study conducted in 21 renal transplant centers in seven countries in which 380 adult primary allograft recipients were randomized within center to receive basiliximab (20 mg i.v.) on days 0 and 4 or placebo in addition to dual immunosuppression with Neoral and steroids. Key clinical events included primary hospitalization, immunosuppressive drug use, patient and graft survival, graft rejection, treatment of rejection, dialysis, and repeat hospitalization. Health resources were valued via a comprehensive electronic cost dictionary, based upon a detailed economic evaluation of renal transplantation in Canada. Medication costs were calculated from hospital pharmacy acquisition costs; basiliximab was assessed a zero cost. RESULTS: The average estimated cost per patient for the first year after transplant was $55,393 (Canadian dollars) for placebo and $50,839 for basiliximab, rising to $141,690 and $130,592, respectively, after 5 years. A principal component of the cost in both groups was accrued during the initial transplant hospitalization ($14,663 for standard therapy and $14,099 for basiliximab). An additional $15,852 and $14,130 was attributable to continued care, graft loss, and dialysis in the two groups, whereas follow-up hospitalization consumed an additional $15,538 for placebo and $13,916 for basiliximab. The mean incremental cost of dialysis was $5,397 for placebo compared with $3,821 for basiliximab, whereas incremental costs of graft loss were $2,548 compared with $2,295 in the two treatment groups. The principal costs associated with repeat admission to the transplant ward and the general ward were marginally higher for placebo ($7,395 vs. $6,300 and $5,986 vs. $4,625). Treatment of acute rejection and maintenance immunosuppressive drug use were associated with only limited savings as a result of basiliximab (savings <$200 each). Sensitivity analysis indicated that the most influential parameters affecting the savings as a result of using basiliximab were a reduction in the duration of initial and repeat hospitalization followed by the reduced risks of acute rejection and graft loss. CONCLUSIONS: Before accounting for the cost of the therapy itself, basiliximab produces an estimated economic saving of $4,554 during the first year after transplant, of which $3,344 is attributable to the reduced costs of graft dysfunction, including graft loss and dialysis ($1,722) and follow-up hospitalizations ($1,622). When marketed, basiliximab is expected to cost approximately $3,000 per course (two doses of 20 mg), resulting in a net first-year saving of $1,554. Under these circumstances, basiliximab can be considered a dominant therapy in renal transplantation. PMID- 11435968 TI - Sirolimus rescue therapy for refractory rejection in renal transplantation. AB - BACKGROUND: Acute renal allograft rejection episodes refractory to antilymphocyte preparations almost inevitably progress to transplantation loss. To reverse ongoing rejection processes, we administered sirolimus (RAPA) after failure of conventional immunosuppressive regimens including full courses of antilymphocyte sera. METHODS: All 36 renal transplantation recipients reported herein displayed either Grade IIB or Grade III biopsy-proven (Banff 1993 criteria) ongoing rejection episodes despite prior treatment with pulse and/or oral recycling of steroids and at least one 14- to 21-day course of murine (OKT3) or equine (ATGAM) antilymphocyte treatment. We compared the actual 12-month outcomes of two demographically similar cohorts of patients treated for refractory rejection with RAPA (Group I; n=24) or mycophenolate mofetil (MMF; Group II; n=12) added to a baseline regimen of cyclosporine (CsA)/prednisone (Pred). RESULTS: Rescue therapy reversed the renal dysfunction in 96% of patients in the RAPA group versus 67% in the MMF group (P=0.03) despite the fact that a greater fraction of patients in the RAPA (17 of 24) than the MMF group (6 of 12) had experienced two or more episodes of acute rejection before study entry and the fact that the recurrent bouts of acute rejection occurred within the first 6 months posttransplant in 94% of patients in the RAPA group compared with 50% (P=0.005) in the MMF group. Among the patients who were reversed successfully, the rates of rebound acute rejection were similar (4% vs. 8%). The mean serum creatinine values were slightly, although not significantly, lower among RAPA than MMF patients at 1, 3, 6, and 12 months: namely, 2.6 vs. 2.8, 2.8 vs. 3.2, 3.0 vs. 3.3, and 2.8 vs. 3.2 mg/dL, respectively. The 1-year patient and graft survival rates were similar: namely, 88% vs. 92% and 83% vs. 67% for the RAPA versus MMF groups. CONCLUSION: RAPA is a potent immunosuppressive agent for the treatment of refractory renal allograft rejection. PMID- 11435969 TI - Effect of low dose cyclosporine and sirolimus on hepatic drug metabolism in the rat1. AB - BACKGROUND: We examined the effect cyclosporine (CsA) and sirolimus (SRL) alone and in combination on hepatic cytochrome P450-mediated metabolism in rats. METHODS: Rats were given 1 mg/kg of CsA or 0.4 mg/kg of SRL alone or in combination via constant intravenous infusion. Renal function was evaluated at the end of treatment. Blood samples were obtained to estimate CsA and SRL concentrations. Hepatic microsomes were prepared for immunoblotting and catalytic assays. RESULTS: CsA alone did not alter serum creatinine levels. SRL given alone or in combination with CsA produced a significant increase in urine output without changes in fluid balance. Although CsA and SRL administered alone caused damage to renal proximal tubules, the two-drug combination dramatically increased the renal structural damage. CsA alone suppressed cytochrome P450 (CYP) 3A2 protein levels by 39% (P=0.012) and catalytic activity by 30% (P=0.042). SRL alone reduced catalytic activity by 38% (P=0.012). Combination therapy reduced both CYP3A2 levels by 55% (P<0.001) and catalytic activity by 55% (P=0.001). CYP2C11 protein expression or catalytic activity were not changed in any group. CYP2A1 protein expression and catalytic activity were both significantly reduced in rats given CsA or/and SRL. Steady-state CsA levels were increased during concurrent SRL dosing, however, SRL concentrations were not changed by CsA coadministration. CONCLUSIONS: Concurrent SRL dosing increases CsA concentrations due to inhibition of hepatic CYP3A2 protein expression. Nephrotoxicity caused by combination therapy is due to CsA elevating levels of SRL or by SRL itself. Concurrent administration of CsA and SRL in transplant patients should be performed with caution. PMID- 11435970 TI - Delayed hepatic artery thrombosis in adult orthotopic liver transplantation-a 12 year experience. AB - BACKGROUND: Although the clinical features of early hepatic artery thrombosis (HAT) are well defined, the features of delayed (more than 4 weeks after transplantation) hepatic artery thrombosis are less clearly defined. The aim of our study was to identify risk factors, clinical presentation, and outcome of management of delayed hepatic artery thrombosis (HAT) after liver transplant (LTx). METHODS: An analysis of prospectively collected data of all patients transplanted from 1986 to 1998 was performed. The importance of recipient (age, sex, primary indication for LTx, cytomegalovirus status, and intraabdominal sepsis) and donor factors (donor age, cold ischemia time, and donor cytomegalovirus status), modes of presentation, and outcome of treatment (biliary reconstruction/stenting, regraft, vascular reconstruction, observation) were analyzed. RESULTS: Delayed HAT was seen in 31/1097 adult LTx recipients (incidence 2.8%). No recipient or donor factors were identified as risk factors. A total of 16 patients were symptomatic at presentation (HAT diagnosed on abdominal ultrasound). Six patients had recurrent episodes of cholangitis, four had cholangitis with a stricture, four had cholangitis and intrahepatic abscesses, and two had bile leaks. Biliary reconstruction was done in six patients (all of whom subsequently required a regraft), vascular reconstruction was performed in two patients (one regrafted and one died shortly after), four patients with cholangitis and stricture on presentation had a biliary stent (all four were later regrafted). A total of 16 patients were regrafted, 9 are alive, 5 died within 6 months (septic at time of LTx), 1 died after 1 year, and 1 died after 2 years. Fifteen patients were asymptomatic and detected on routine screening. 5 have remained asymptomatic and are still alive, 1 developed a biliary stricture that was stented and is alive 105 months later, 4 had recurrence of the original disease, 3 developed progressive graft failure and were listed for transplant but died before regraft due to overwhelming sepsis and hepatic encephalopathy. Two patients died due to nonbiliary sepsis. CONCLUSIONS: Delayed HAT is a rare complication of LTx that may present with biliary sepsis, or remain asymptomatic. Biliary or vascular reconstructions do not increase graft survival. Of the patients who were clinically silent on presentation, 20% developed progressive graft failure requiring a second transplant. A total of 33% survived in the long-term without a second transplant. Ongoing severe sepsis at the time of regraft results in poor survival. PMID- 11435971 TI - The long-term effect of simultaneous heart and kidney transplantation on native renal function. AB - BACKGROUND: It is unclear whether patients with heart failure and renal insufficiency should receive a simultaneous heart and kidney transplant or whether a single heart transplantation is sufficient to restore native renal function. METHODS: We analyzed the renal plasma flow and glomerular filtration of the native and transplant kidneys in eight patients long term after simultaneous heart and kidney transplantation using a dynamic MAG3 radioisotope scan and serum creatinine determinations. All subjects had been hemodialysis dependent before transplantation. Seven patients suffered from an intrinsic renal disease that were diabetic nephropathy in three cases, small fibrotic kidneys of undetermined origin in two cases, one lupus nephritis, and cyclosporine nephrotoxicity in one patient who had a previous heart transplant. In one patient renal insufficiency was considered to be solely due to renal hypoperfusion because no intrinsic renal disease could be detected. RESULTS: All patients were on cyclosporine-based triple immunosuppression, transplanted for 4 to 10 years, exhibited cardiac ejection fractions of more than 50% and had normal serum creatinine values. Radioisotopic scan showed no function of the native kidneys in all seven patients with intrinsic renal disease but exhibited normal function of the native kidneys as well as the renal transplant in the patient without intrinsic kidney disease before transplantation. CONCLUSIONS: These data suggest that a simultaneous heart and kidney transplantation is necessary in patients with cardiomyopathy and renal insufficiency due to primary kidney disease, but not in those with hemodynamically mediated renal failure, even if an immunosuppressive regimen with calcineurin inhibitors is used. PMID- 11435972 TI - Mechanisms of thrombotic microangiopathy following xenogeneic hematopoietic progenitor cell transplantation. AB - INTRODUCTION: Attempts to induce tolerance though mixed hematopoietic chimerism in the discordant pig-to-baboon xenotransplantation model are sometimes complicated by a potentially fatal thrombotic microangiopathy in the recipient baboons. This state develops immediately after the infusion of porcine mobilized peripheral blood leukocytes, containing progenitor cells (PBPC). In our study, we examined the interaction of infused porcine PBPC with recipient platelets in vivo in baboons and investigated the underlying mechanisms using an in vitro model. METHODS: Two naive baboons and six baboons preconditioned with irradiation and immunosuppression that received porcine PBPC were evaluated in vivo. The interaction of porcine and baboon PBPC with baboon platelets was investigated by an in vitro platelet aggregation assay. Fresh and cryopreserved PBPC were evaluated as well as PBPC obtained from growth-factor mobilized and unmobilized pigs. Furthermore, cellular subsets of PBPC were assessed for potential to induce platelet aggregation. Immunohistochemical staining was performed on platelet leukocyte aggregates and potential inhibition of aggregation with anti-P-selectin and anti-CD154 mAbs, or eptifibatide (a GPIIb/IIIa receptor antagonist), was tested. RESULTS: All baboons that received porcine PBPC rapidly developed marked thrombocytopenia (<20,000/microl), elevated serum lactate dehydrogenase (>1,500U/liter), schistocytosis, and platelet aggregates on blood smear. Three baboons died (two untreated and one preconditioned), and substantive platelet aggregates containing porcine leukocytes were observed in the microvasculature of lungs and kidneys. In vitro, porcine, but not baboon, PBPC induced aggregation of baboon platelets in a dose-dependent manner. Immunohistological examination of these aggregates confirmed the incorporation of porcine leukocytes. Cryopreserved PBPC caused less aggregation than fresh PBPC, and growth-factor-mobilized PBPC induced less aggregation than unmobilized PBPC. Aggregation was fully abrogated by the addition of eptifibatide, and modulated by anti-P-selectin and anti-CD154 monoclonal antibodies that recognize adhesion receptors on activated platelets. Purified fractions (granulocytes, CD2+, and CD- cells) of porcine PBPC did not initiate aggregation, whereas addition of exogenous porcine PBPC membranes (erythrocytes, dead cells, and/or platelets) to the purified fractions exacerbated the aggregation response. CONCLUSIONS: These data indicate that porcine PBPC mediate aggregation of baboon platelets. This process likely contributes to the thrombotic microangiopathy observed after PBPC transplantation in the pig-to-baboon model. Eptifibatide can fully abrogate platelet aggregation induced by porcine PBPC in vitro. Purification of the progenitor cells from porcine PBPC and/or treatment of baboons with eptifibatide may be beneficial. PMID- 11435973 TI - Evaluation of CMV viral load using TaqMan CMV quantitative PCR and comparison with CMV antigenemia in heart and lung transplant recipients. AB - BACKGROUND: Quantitative assessment of cytomegalovirus (CMV) infection using the antigenemia test has been used to monitor CMV infection in heart and lung transplant patients enabling a preemptive treatment strategy. However, the method is labour intensive, samples have to be processed within a few hours and requires skilled interpretation. A comparative prospective evaluation of a real-time TaqMan CMV quantitative PCR (QPCR) with the CMV antigenemia was undertaken. METHODS: A real-time quantitative TaqMan CMV PCR from EDTA bloods was developed. In this study 25 heart transplant and single-lung transplant patients were monitored posttransplantation by antigenemia and TaqMan CMV QPCR. CMV DNA extracted from EDTA blood was amplified by TaqMan QPCR using primers and probe designed from the CMV glycoprotein B (gB) gene. Quantification of the genome copies is extrapolated from a standard curve generated from amplification of quantified standards. RESULTS: Antigenaemia levels and TaqMan CMV QPCR genome copies showed a linear correlation between the two assays (R=0.843, P=0.001). A clinically significant threshold of 50 CMV pp65 antigen positive polymorphonuclear leucocytes (PMNLs) per 200 000 cells previously reported was used to extrapolate an equivalent value of 40 000 (log 4.6) genome copies per ml of blood for the TaqMan CMV QPCR. CONCLUSIONS: The TaqMan system enables a rapid high-throughput of samples. The TaqMan CMV QPCR can be used as an accurate and robust alternative to the antigenemia test to predict CMV disease and to monitor effectiveness of treatment. PMID- 11435974 TI - Prolonged inhibition of obliterative airway disease in murine tracheal allografts by brief treatment with anti-leukocyte function-associated antigen-1 (CD11a) monoclonal antibody. AB - BACKGROUND: We have previously shown that anti-leukocyte function-associated antigen (LFA)-1 (CD11a) monoclonal antibody (mAb) prevents acute rejection and produces donor-specific unresponsiveness in murine recipients of heterotopic heart allografts. Here, we investigate the ability of this mAb to prevent the development of obliterative airway disease (OAD) in murine recipients of tracheal allografts. METHODS AND RESULTS: BALB/c tracheae were heterotopically transplanted into C3H mice. OAD developed by day 28 after transplantation and was characterized histologically by a loss of epithelial cell coverage and luminal obliteration of the tracheal allograft with a proliferation of fibrogenic mesenchymal cells, which is a lesion comparable to obliterative bronchiolitis in human lung transplant recipients. Monotherapy with anti-LFA-1 mAb preserved graft epithelium, prevented the development of OAD, and maintained unresponsiveness to donor antigen for more than 42 days after the final mAb administration. CONCLUSION: These findings suggest the potential for anti-LFA-1 mAb therapy to suppress both acute and chronic rejection in clinical lung transplantation. PMID- 11435975 TI - Human T cells infiltrate and injure pig coronary artery grafts with activated but not quiescent endothelium in immunodeficient mouse hosts. AB - BACKGROUND: We have previously demonstrated that human artery grafts transplanted to immunodeficient mice are infiltrated and injured by unsensitized allogeneic human T cells. We extended our investigations to human anti-porcine xenoresponses in this model. METHODS: Pig coronary artery segments were interposed into the infrarenal aorta of severe combined immunodeficiency/beige mice. After 7 days, certain recipients were reconstituted with human leukocytes and/or treated with proinflammatory cytokines. The grafts were harvested after 1-70 days and examined by histology, immunohistochemistry, and morphometry. RESULTS: Pig artery grafts from untreated mice had no evidence of injury, leukocytic infiltrate, or endothelial cell activation up to 70 days postoperatively, despite deposition of murine complement. Host reconstitution with human peripheral blood mononuclear cells resulted in a discrete population of circulating T cells that did not infiltrate or injure the grafts up to 28 days after adoptive transfer. Administration of porcine interferon-gamma for up to 28 days sustained the expression of graft vascular cell adhesion molecule-1 and major histocompatibility complex antigens, but did not initiate recruitment of human leukocytes. In contrast, treatment with human tumor necrosis factor for 7 days induced the de novo expression of porcine E-selectin by graft endothelial cells and elicited human T cell infiltration and human peripheral blood mononuclear cell-dependent vascular injury. CONCLUSIONS: The human peripheral blood mononuclear cell-severe combined immunodeficiency/beige mouse model identifies a significant difference between human T cell allogeneic and xenogeneic responses in vivo. Xenografts with quiescent endothelium are not infiltrated or injured by T cells under the same conditions in which allografts are rejected. Activation of pig coronary artery endothelial cells by human tumor necrosis factor, but not porcine interferon-gamma, elicits cellular xenoresponses. PMID- 11435976 TI - Xenogeneic extracellular matrix grafts elicit a TH2-restricted immune response. AB - BACKGROUND: Porcine small intestinal submucosa (SIS) is an acellular, naturally derived extracellular matrix (ECM) that has been used for tissue remodeling and repair in numerous xenotransplantations. Although a vigorous immune response to xenogeneic extracellular matrix biomaterials is expected, to date there has been evidence for only normal tissue regeneration without any accompanying rejection. The purpose of this study was to determine the reason for a lack of rejection. METHODS: Mice were implanted s.c. with xenogeneic tissue, syngeneic tissue, or SIS, and the graft site analyzed histologically for rejection or acceptance. Additionally, graft site cytokine levels were determined by reverse transcriptase polymerase chain reaction and SIS-specific serum antibody isotype levels were determined by ELISA. RESULTS: Xenogeneically implanted mice showed an acute inflammatory response followed by chronic inflammation and ultimately graft necrosis, consistent with rejection. Syngeneically or SIS implanted mice, however, showed an acute inflammatory response that diminished such that the graft ultimately became indistinguishable from native tissue, observations that are consistent with graft acceptance. Graft site cytokine analysis showed an increase in interleukin-4 and an absence of interferon-gamma. In addition, mice implanted with SIS produced a SIS-specific antibody response that was restricted to the IgG1 isotype. Reimplantation of SIS into mice led to a secondary anti-SIS antibody response that was still restricted to IgG1. Similar results were observed with porcine submucosa derived from urinary bladder. To determine if the observed immune responses were T cell dependent, T cell KO mice were implanted with SIS. These mice expressed neither interleukin-4 at the implant site nor anti SIS-specific serum antibodies but they did accept the SIS graft. CONCLUSIONS: Porcine extracellular matrix elicits an immune response that is predominately Th2 like, consistent with a remodeling reaction rather than rejection. PMID- 11435977 TI - CD4+ T cell recognition of a single discordant HLA-A2-transgenic molecule through the indirect antigen presentation pathway induces acute rejection of murine cardiac allografts. AB - To further define the role of indirect allorecognition, cardiac allografts from HLA-A2-transgenic (HLA-A2+) C57BL/6 mice were heterotopically transplanted into normal C57BL/6, CD4 T cell-knockout (KO) C57BL/6 mice, CD8 T cell-KO C57BL/6 mice, fully MHC-discordant BALB/c mice (allogeneic control), and HLA-A2+ C57BL/6 mice (syngeneic control). HLA-A2+ grafts were acutely rejected when transplanted into BALB/c mice (mean survival time: 10+/-0.8 days), normal C57BL/6 mice (mean survival time: 16.5+/-2.1 days) as well as CD8-KO mice (mean survival time: 12.8+/-1.3 days). Histopathological analysis revealed classical acute cellular rejection with moderate to severe diffuse interstitial CD4+ and CD8+ cellular infiltrates and significant intra-graft deposition of IgG and complement. In contrast, HLA-A2+ grafts were not rejected when transplanted into CD4-KO mice or HLA-A2+ mice. CD8-KO recipients treated with an anti-CD4 monoclonal antibody, but not with an anti-NK monoclonal antibody, failed to reject their allografts with prolonged administration of antibody (30 days). Spleen cells from mice rejecting HLA-A2+ allografts failed to lyse HLA-A2+ target cells indicating a lack of involvement of CD8+ T cells in the rejection process. In contrast, spleen cells from rejecting animals proliferated significantly to both HLA-A2+ cells and to a peptide derived from the HLA-A2 molecule. Development of anti-HLA-A2 antibodies was observed in all animals rejecting HLA-A2+ allografts. These results suggest that indirect allorecognition of donor MHC class I molecules leads to rejection of cardiac allografts and development of alloantibodies in this unique transplant model in which there is a single MHC discordance between donor and recipient. PMID- 11435978 TI - Rantes production during development of cardiac allograft vasculopathy. AB - BACKGROUND: RANTES (regulated on activation, normal T cell expressed and secreted) production has been shown to correlate with mononuclear cell recruitment and precede intimal thickening in cardiac allograft vasculopathy (CAV). However, the cells that produce RANTES in CAV are undefined. Therefore, in an MHC II-mismatched murine model of CAV, we sought to (1) define the cellular sources of RANTES and (2) determine the role of CD4+ lymphocytes in RANTES production during CAV development. METHODS: B6.CH-2bm12 strain donor hearts were transplanted heterotopically into wild-type (WT) or CD4 knockout (CD4KO) C57BL/6 mice (MHC II mismatch). No immunosuppression was used. Recipients were sacrificed at 7, 14, and 24 days. Intragraft RANTES gene expression and protein levels were determined with ribonuclease protection assay and ELISA, respectively. At days 7 and 24, RANTES production by graft-infiltrating cells was defined with intracellular RANTES staining and multicolor FACS analysis. Intimal thickening was quantitated morphometrically. In murine hearts and in six explanted human hearts with advanced CAV, RANTES was also localized immunohistochemically. RESULTS: NK, NKT, and gammadelta+ cells, in addition to CD4+, CD8+ lymphocytes, and CD11b+ macrophages, produced RANTES in early and late stages of CAV. RANTES producing NK, NKT, and gammadelta+ cells tripled in number during CAV development; by day 24, NK and gammadelta+ cells each outnumbered CD4+ lymphocytes and CD11b+ macrophages. The presence of CD4+ lymphocytes was required for sustained RANTES production in allografts, which correlated with mononuclear cell recruitment and preceded intimal thickening. In murine and explanted human hearts with advanced CAV, RANTES immunolocalized with graft-infiltrating mononuclear cells and vessel wall cells. CONCLUSIONS: We present evidence that other cell types in addition to CD4+, CD8+ T lymphocytes, and CD11b+ macrophages contribute significantly to RANTES production in CAV. In this MHC II-mismatched murine model of CAV, sustained RANTES production requires CD4+ lymphocytes, correlates with mononuclear cell recruitment, and precedes intimal thickening. In experimental and human CAV, vessel wall cells may also produce RANTES. Interventions aimed at inhibiting RANTES production in CAV may need to target several types of cells, and neutralization of RANTES bioactivity may reduce mononuclear cell recruitment and CAV development. PMID- 11435979 TI - Immunotherapy with nondepleting anti-CD4 monoclonal antibodies but not CD28 antagonists protects islet graft in spontaneously diabetic nod mice from autoimmune destruction and allogeneic and xenogeneic graft rejection. AB - BACKGROUND: T-cell activation and the subsequent induction of effector functions require not only the recognition of antigen peptides bound to MHC molecules by T cell receptor (TCR) for antigen but also a costimulatory signal provided by antigen presenting cells. CD4 T-cell activation and function require the CD4 molecule as a coreceptor of TCR. The CD28/B7 pathway is a major costimulatory signal for T-cell activation and differentiation. METHODS: The effect of targeting CD4 by nondepleting anti-CD4 monoclonal antibodies (mAbs) versus blocking CD28/B7 by CTLA4Ig, anti-CD80 mAbs, and anti-CD86 mAbs on the prevention of recurrence of autoimmune diabetes after MHC-matched nonobese diabetes resistant (NOR) islet transplantation in nonobese diabetic (NOD) mice were compared. Whether nondepleting anti-CD4 mAbs prolong allogeneic islet graft survival and xenogeneic pig islet graft survival in diabetic NOD mice were studied. Furthermore, the effect of nondepleting anti-CD4 mAbs combined with CTLA4Ig on allogeneic islet graft survival in NOD mice was investigated. RESULTS: Recurrence of autoimmune diabetes can be prevented by nondepleting anti-CD4 mAbs. Blocking the CD28/B7 costimulatory pathway by CTLA4Ig or by anti-CD80 mAbs and anti-CD86 mAbs cannot prevent recurrence of autoimmune diabetes after islet transplantation. Short-term treatment with nondepleting anti-CD4 mAbs significantly prolongs allogeneic islet graft survival and xenogeneic pig islet graft survival in diabetic NOD mice. But nondepleting anti-CD4 mAbs combined with CTLA4Ig decreased allogeneic islet graft survival. CONCLUSIONS: Nondepleting anti CD4 mAbs but not CD28 antagonists protect islet grafts in diabetic NOD mice from autoimmune destruction and allogeneic and xenogeneic graft rejection. The efficacy of nondepleting anti-CD4 mAbs is compromised when it combines with CTLA4Ig. PMID- 11435980 TI - The contribution of adhesion molecule expression in donor kidney biopsies to early allograft dysfunction. AB - BACKGROUND: Renal allograft rejection is associated with the expression of adhesion molecules on vascular endothelial and tubular epithelial cells. METHODS: To assess whether the number of cell adhesion molecules expressed in donor kidneys can predict early rejection or delayed graft function, kidney biopsies from 20 living and 53 cadaveric kidney donors were obtained before engraftment into the recipients and the expression of the cell adhesion molecules intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and endothelial leukocyte adhesion molecule (E-selectin) were determined by immunohistochemistry. RESULTS: All biopsies from living donors showed significantly lower expression of ICAM-1 and VCAM-1 compared to biopsies from cadaveric donors. There was no difference in the expression of adhesion molecules on tubular cells between transplants with primary function compared to allografts with early rejection in living donated kidneys (ICAM-1: 2+/-8 vs. 3+/ 8%; VCAM-1: 9+/-7 vs. 1+/-1%), as well as in cadaveric kidneys (ICAM-1: 38+/-29 vs. 39+/-38%; VCAM-1: 55+/-27 vs. 48+/-29%). The expression of ICAM-1 molecules on tubular cells was determined to be a predictor for the occurrence of delayed graft function in cadaveric kidneys (ICAM-1: 65+/-24* vs. 38+/-29% delayed graft versus primary graft function). No delayed graft function occurred in recipients of living donated kidneys. CONCLUSIONS: These data suggest that adhesion molecule expression in donor biopsies is not a predictor for early allograft rejection, but can be used as a marker for the development of postischemic acute renal allograft failure. PMID- 11435981 TI - Lowering of blood glucose to nondiabetic levels in a hyperglycemic pig by allografting of fetal pig isletlike cell clusters. AB - BACKGROUND: Fetal pig isletlike cell clusters (ICCs) will differentiate when grafted into the thymus gland of outbred immunosuppressed nondiabetic pigs for up to 3 months. Whether these cells will survive for a similar period in a diabetic recipient and will mature with secretion of insulin to ameliorate the hyperglycemia is unknown. METHODS: Between 40,000 and 125,000 ICCs (7,000 to 11,400 ICCs/kg) were injected into the thymus gland of five juvenile pigs immunosuppressed with cyclosporine and deoxyspergualin, and the animals were subsequently made diabetic by the injection of streptozotocin. Insulin was administered subcutaneously, with one pig dying from hypoglycemia. The animal with the least number of ICCs transplanted was killed 81 days later, and the graft was analyzed histologically. Blood glucose levels and porcine C-peptide in the remaining animals were monitored for a median of 101 days. RESULTS: Histological analysis of the graft showed numerous epithelial cell clusters; the percentage of cells that contained insulin, glucagon, somatostatin, and pancreatic polypeptide were 61%, 64%, 25%, and 18%, respectively. Some cells contained more than one hormone. Porcine C-peptide was detected from 21 days after induction of diabetes but not before. In the pig receiving the most ICCs, blood glucose levels were lowered to nondiabetic levels 109 days after transplantation. Plasma C-peptide levels in response to glucagon in this pig steadily increased after grafting; peak levels were 0, 0.21, 0.45, and 0.52 ng/ml at 4, 21, 49, and 80 days after induction of diabetes compared to 0.09 ng/ml in control diabetic pigs. The secretion of C-peptide in response to oral and intravenous glucose and arginine also was greater than in untransplanted diabetic pigs, the pattern of secretion being consistent with developing fetal beta cells as the source of the C-peptide. Pancreatic insulin content was 0.1 mU/mg, 4% of that in nondiabetic pigs, and the number of beta cells per islet was 3 to 6 compared to 90 in nondiabetic controls. CONCLUSIONS: ICCs will differentiate and function for up to 111 days when transplanted into outbred immunosuppressed pigs rendered diabetic. Blood glucose levels can be lowered to nondiabetic levels when sufficient numbers of ICCs are grafted. PMID- 11435982 TI - Human monocytic ehrlichiosis: an emerging pathogen in transplantation. AB - BACKGROUND: The spectrum of disease caused by Ehrlichia spp. ranges from asymptomatic to fatal. Awareness and early diagnosis of the infection is paramount because appropriate therapy leads to rapid defervescence and cure. If left untreated, particularly in immunosuppressed patients, ehrlichioses may result in multi-system organ failure and death. METHODS: We report the second case of human monocytic ehrlichiosis (HME) in a liver transplant recipient, and review the literature. RESULTS: The patient presented with fever and headache, had negative cultures, and despite broad-spectrum antimicrobial coverage appeared progressively septic. After eliciting a history of tick exposure we treated the patient empirically with doxycycline. The diagnosis of HME was confirmed by 1) polymerase chain reaction (PCR) for Ehrlichia chaffeensis, 2) acute and convalescent serum titers, and 3) in vitro cultivation of E chaffeensis from peripheral blood. CONCLUSION: Although human ehrlichioses are relatively uncommon, they are emerging as clinically significant arthropod-borne infections. Although epidemiological exposure is responsible for infection, immunosuppression makes patients more likely to succumb to disease. A high index of suspicion and early treatment results in a favorable outcome. PMID- 11435983 TI - Tolerance in renal transplantation after allogeneic bone marrow transplantation-6 year follow-up. AB - Despite significant advancements in clinical transplantation, very few reports describe the long-term acceptance of transplanted solid organs without indefinite immunosuppression. The immunosuppressive agents used are nonspecific and have serious potential side effects. We present a patient who received a living-donor renal allograft from the same person who had donated bone marrow to her several years earlier. Tolerance was expected based on previous acceptance of full thickness skin grafts from the donor. Indeed, there has been no evidence of rejection during a 6-year follow-up period, and no induction or maintenance immunosuppression has been given. All noninvasive parameters of graft function remain normal. This and similar reports prove that genetically disparate solid organs can coexist without pharmacological immunosuppression. PMID- 11435985 TI - Reconsideration of HIV as a contraindication to transplantation. PMID- 11435984 TI - Posttransplant lymphoproliferative disorder after liver transplantation in miniature swine. AB - BACKGROUND: Posttransplant lymphoproliferative disorder (PTLD) is a well-known, life-threatening complication of immunosuppressive therapy, occurring in both adult and pediatric transplant recipients. METHODS: To study the effect of major histocompatibility complex on tolerance induction to primarily vascularized liver allograft, a semi-identical miniature swine model was developed to mimic the clinical situation of parent-into-infant liver transplantation. Long-term acceptance of semi-identical liver allograft was obtained by a transient course of FK506. In a subgroup of six animals, three developed a lethal PTLD. These animals were studied by histology and immunohistochemistry and the anti-donor cellular immune response was assessed. In addition, the possible viral origin of the proliferative process was evoked. RESULTS: Histology and immunohistochemistry revealed an abnormal B-cell proliferation in many organs of swine suffering from PTLD. Evidence of human Epstein-Barr virus, cytomegalovirus, and adenovirus was not evidenced, but a porcine virus responsible for a respiratory and reproductive syndrome (PRRS) was identified in the lymphoid tissue of these animals. In mixed lymphocyte reaction, a significant antiself immune response confirmed an infection by a virus. CONCLUSIONS: This is the first report suggesting that PRRS virus might provoke PTLD in immunosuppressed miniature swine after orthotopic liver transplantation. Whether PTLD could be induced by injection of the PRRS virus in immunosuppressed animals, a pig model of PTLD might be developed and would represent an interesting preclinical model for testing anti-PTLD therapies. PMID- 11435986 TI - Living anonymous kidney donation: what does the public think? AB - BACKGROUND: Health professionals are increasingly turning to living organ donation to augment cadaveric donation. Although living donation is currently performed with donors who are either genetically or emotionally related to the recipient, a 1997 British Columbia Transplant Society survey indicated that 32% of BC residents would be willing to donate a kidney, while alive, to a stranger (unpublished data). The goal of this study is to tap the public pulse about the living anonymous donor (LAD) by replicating and expanding the 1997 findings. METHODS: Five hundred BC residents completed a telephone survey including demographic information, questions about their organ donation behaviors and attitudes, and their willingness to donate a kidney, while alive, to particular individuals (child, spouse, parent, relative, friend, and stranger). To improve the methodological rigor of the 1997 study, an informed condition was added in the current study where participants learned about living donation before being asked about their willingness to donate. RESULTS: There were no differences among the 1997 results and the two conditions in the 2000 survey. Twenty-eight percent of participants in the uninformed condition and 29% of participants in the informed condition indicated that they would be willing to be LADs. LADs were more likely than self-reported non-donors to have registered as cadaveric donors and to endorse attitudes that were congruent with wanting to donate to a stranger. CONCLUSIONS: This study replicates the 1997 findings and increases confidence that a significant minority of British Columbians support living anonymous donation and that some would consider becoming LADs themselves. PMID- 11435987 TI - Case 1: persistent acute rejection after kidney transplantation. PMID- 11435988 TI - [Realities and paradoxes]. PMID- 11435989 TI - [Reduction in the number of lethal heroin overdoses in France since 1994. Focus on substitution treatments]. AB - BACKGROUND: Since 1994-1995, rapid development of widely available substitution treatments has appeared to be a major healthcare step in heroin addiction. Currently approximately 60000 patients are taking daily maintenance doses of oral methadone and about 7200 are taking sublingual buprenorphine. In parallel with the expansion of these treatments, the number of lethal overdoses has fallen off regularly: 564 in 1994, 393 in 1996 and 143 in 1998 (-74.6% in 4 years). AIM OF THE STUDY: We searched for a correlation between the rise in the number of patients taking maintenance treatments and the decreased in recorded deaths due to heroin overdose. Other factors which may influence this decrease were also considered. RESULTS AND DISCUSSION: A linear correlation was found between the increasing number of patients on maintenance treatment (high-dose buprenorphine or methadone) and the decrease in fatal heroin overdoses in France between 1994 and 1998. The importance of this correlation must be modulated by the presence of other events such as political, social, healthcare and behavioral events concerning drug users. PMID- 11435990 TI - [Referral of drug addicts "in crisis" seen in the emergency room]. AB - This survey was conducted to estimate the number of drug addicts seen in emergency care units in the Paris hospitals for a drug-related crisis and to analyze subsequent dispatching decisions made over a 14-day period. Twelve polyvalent emergency units participated in the study. These units cared for 116 drug abusers in a crisis situation during the 14-day study period. After initial assessment, the emergency care physician proposed a transfer center in 74% of the cases. Proposed referral centers included: an ECIMUD (Coordination and medical care team for drug addicts) for 27% of the patients, a psychiatric outpatient clinic for 33%, a CSST (Specialized center for drug abusers) for 2% and hospitalization for 11%. After a second assessment by a psychiatrist or an ECIMUD physician, 53% of the patients with a second assessment were referred to an ECIMUD, 32% to a psychiatric outpatient clinic, and 6% to a CSST; 26% were hospitalized. This survey points out the eminent role of psychiatric outpatient clinics and ECIMUD teams in providing emergency care for drug abusers. PMID- 11435991 TI - [Psychoactive drug use in a declared non-addicted control sample and comorbidity. Results of a study in 860 French-speaking subjects]. AB - AIMS: This study, conducted within the framework of a broader research program of the INSERM 494013 Dependence Network, was designed to estimate illicit drug use and tobacco smoking in a declared non-addicted sample and to determine whether illicit drug users differ from non-users in terms of comorbidity. METHODS: The study was conducted in an "all and sundry" sample of subjects. Patterns of drug use and comorbid factors (psychiatric disorders, suicide attempts, repeated accidents, social inadaptation) were assessed using a semi-structured interview (heteroevaluation, MINI DSM IV interview, Groningen). RESULTS: Among 860 subjects, 107 (12.4%) used illicit drugs and 26 of these 107 (24.3%) were dependent users or abusers. Specific analysis of non-dependent non-abuser subjects who had used illicit drugs (70 occasional and 11 regular users) showed a higher rate of use in younger subjects (12.7% in the 15-24 year group, 5.7% in the 24-49 year group) and men. Except for repeated accidents (OR=5.5 [1.6-18.5]), comorbid disorders were not more frequent in non-users than in users. CONCLUSION: Besides use for recreational purposes, the rate of use of illicit drugs with abuse or dependence was high in our non-clinical sample. Although no specific comorbid psychiatric disorders were identified among non-dependent non-abuser subjects who had used illicit drugs, the frequency of repeated accidents evidenced the ill-fated side effects of illicit drugs and/or the specific biopsychological vulnerability of these subjects. This highlights the importance of not neglecting drug abuse. PMID- 11435992 TI - [Two-year follow-up of an opioid-user cohort treated with high-dose buprenorphine (Subutex)]. AB - AIM OF THE STUDY: Care for opioid users changed greatly in France in 1996 when general practitioners (GP) were allowed to prescribe high-dose sublingual buprenorphine (Subutex((R))) for maintenance treatment of major opioid dependence. In order to evaluate treatment benefits, a prospective epidemiological 2-year follow-up was initiated in May 1996 with the participation of 105 French GPs. METHODS: A cohort of outpatient opioid users who started high dose sublingual buprenorphine maintenance therapy at study onset or who had recently started were included in a prospective epidemiological study by GPs involved in management of drug abusers. Patients were followed for 2 years with collection of standardized information at 1, 3, 6, 12, and 24 months. The main evaluation criteria were follow-up by the same GP throughout the study and retention in the care system 2 years later. For patients who fulfilled these criteria, secondary end points were analyzed: information about buprenophine prescription, social status, and hepatitis B and C and HIV seroconversions. RESULTS: The 101 GPs included 919 patients and 909 were analyzed 2 years later. At study onset, a majority of the patients (70.6%) were taking an ongoing maintenance treatment, 10.5% had previously received such a treatment and the treatment was initiated for 18.8%. At the end of the study, 508 patients (55.9%) were still being followed by the same GP and 101 (11.1%) were followed by another healthcare provider (another GP, hospital or specialized center). No information about the care giver was available for 82 patients (9%). Among the other patients, 123 (13.5%) were lost to follow-up, 24 (2.6%) had moved, 23 (2.6%) were incarcerated, 11 (1.2%) had successfully discontinued drug usage and 7 (0.8%) had died. Other reasons for unsuccessful follow-up by the same GP were mainly (for 6 patients each): relapse, switch to methadone, no medical information, non compliance with scheduled controls. Among the patients followed by the same GP, declaration of heroin and drug intake significantly decreased (p<0.001), and social status (GAF scale) and TMSP evaluation significantly improved (p<0.001). The social situation (housing condition and work) also improved significantly (p<0.001). The rate of buprenorphine treatment was 84% with longer and less fractionated prescriptions. The HBV, HBC and HIV seroconversion rates were low in this high-risk population (2.7%, 4.1% and 0.8% respectively). CONCLUSION: This two-year follow-up of 909 opioid users showed that nearly 70% of the patient remained within the healthcare system, mainly with the same GP or more rarely with another practitioner. Among the 508 patients still followed by the same GP, maintenance treatment with high-dose buprenorphine was observed in more than 80% of the patients. These patients had a significantly improved social status, a significant decrease in drug intake and a significant improvement in their social adaptation and severity of drug abuse. PMID- 11435993 TI - [Prevalence of cannabis and tobacco use in patients attending a methadone center. Evaluation by urinary biomarkers]. AB - AIM OF THE STUDY: The aim of this study was to determine the prevalence of cannabis and tobacco use in a population attending a Methadone Center. METHODS: We studied 49 patients over a 3-month period. Urine samples were obtained at three different times from each patient for analysis. We measured urinary cannabinoids using an immunopolarization assay to identify cannabinoid use. Cotinine concentrations were quantified in urine using a colorimetric assay to assess total urinary nicotine metabolites. RESULTS: The prevalence of cannabis use was high in this population. Urinary cannabinoids were present in 63% of the samples. Similarly 90% had high-urinary cotinine concentrations. CONCLUSION: Measurement of urinary biomarkers showed that tobacco and cannabis use was high among patients attending a Methadone Center. PMID- 11435994 TI - [Neuropsychic effects of dehydroepiandrosterone]. AB - Dehydroepiandrosterone (DHEA) and DHEA sulfate (DHEA-S) are secreted primarily by the adrenal glands. DHEA could also be a neuroactive steroidal hormone. Because basal levels of DHEA and DHEA-S in humans decrease significantly with age, these hormones have been assumed to be involved in the aging process and in a number of pathologies which develop with aging: immunosenescence, increased mortality, increased incidence of cancer, osteoporosis and cardiovascular diseases. However, its role is still unknown. In humans, cross sectional and longitudinal studies have shown that DHEA might be associated with global measures of well-being and functioning, but positive effects on measures of memory and attention could not be found. Studies investigating DHEA and DHEA-S levels in dementia have produced controversial results. Short-term experimental studies have not shown significant improvement in global measures of well-being and functioning in healthy subjects but have revealed preliminary evidence for mood enhancing and antidepressant effects of DHEA. There is no evidence that DHEA could induce addiction in human beings. PMID- 11435995 TI - [Analgesic-induced chronic headache]. AB - Analgesic-induced chronic headaches are the outcome of an overtreatment of migraine or tension type headaches. Progressively, the usual headache changes and becomes a chronic daily headache occurring in the morning when waking up, relieved 1 hour after intake of the concerned analgesic. If drug intake is delayed, the daily headache disappears after 4 weeks following a period of worsening. This withdrawal therapy is often conducted during a 2-week hospitalization. Drugs mainly implicated in this type of dependence are: caffeine, ergotamine, simple analgesics, and triptans which are becoming more and more involved. The best treatment is prevention. When first seen, patients must be warned of the risk of chronic use of analgesics. PMID- 11435996 TI - [Causality and psychopathological course in a cannabis dependency case]. AB - We analyzed the behavior of teenagers who use cannabis in order to identify the drive leading to cannabis abuse and the perceived benefit which apparently sustains dependency. This analysis pointed out the following issues: identification, alterity modulation, modification of thinking activity, pursual of a psychopathological state of deterioration. This short clinical dissertation should be completed by further more deeply oriented clinical research. PMID- 11435997 TI - Involvement of free radicals in MDMA-induced neurotoxicity in mice. AB - 3,4-methylenedioxymethamphetamine (MDMA) or ecstasy) is a substituted amphetamine with stimulating and hallucinogenic properties. Administration of MDMA leads to the formation of metabolites responsible for its toxic effects on serotonergic neurons in rats and non-human primates and on dopaminergic neurons in mice. Our findings indicate that overexpression of the human superoxide dismutase gene (Cu/Zn-SOD) abolishes certain effects of MDMA such as the decreased level of dopamine, DOPAC and 5-HT in the striatum, inactivation of certain antioxidant enzymes (CU/ZN-SPD, catalase or glutathione peroxidase) or peroxidation of lipids. These data are in agreement with the implication of free radicals and consequenty of oxidative stress in the mode of action of MDMA. PMID- 11435998 TI - Multilamellar bodies as potential scattering particles in human age-related nuclear cataracts. AB - PURPOSE: To characterize within human age-related nuclear cataracts rare spherical objects covered by multiple membranes, termed multilamellar bodies (MLBs). METHODS: Adult human normal, transparent lenses were obtained from eye bank donors and age-related nuclear cataracts were obtained immediately after extracapsular extraction. Each sample was Vibratome sectioned fresh into 200 microm thick sections that were fixed and embedded for light or electron microscopy. Confocal images were recorded from sections stained with the lipid soluble dye, DiI. RESULTS: Light micrograph montages of the equatorial plane containing the fetal and embryonic nuclei were examined. Rare, but distinct, circular 1-3 microm diameter objects were observed consistently in the cataracts. These objects did not appear to be components of the complex intercellular interfaces. Serial sections indicated that the objects were spherical, or contained a spherical component. For about 20,000 fiber cell cross-sections in each lens, the frequency of MLBs was 10 times higher in cataracts than in the normal lens nuclei. Although extensive searching with the electron microscope was necessary, the size, circular profile and multiple layers of thin (5 nm) membranes easily identified the MLBs. Interiors of the MLBs displayed variable textures. Confocal images indicated that the coverings were enriched in lipid compared to the adjacent plasma membranes. The calculated density of the MLBs in the cataractous nuclei was about 3800/mm3, which represents a volume fraction of 0.00003. CONCLUSIONS: Because the MLBs are large compared to the wavelength of light, display interiors with variable staining textures and have lipid-rich coverings, they appear to be ideal candidates for large scattering particles that may contribute to the forward light scattering in nuclear cataracts. PMID- 11435999 TI - Subretinal injections in rodent eyes: effects on electrophysiology and histology of rat retina. AB - PURPOSE: To describe a reliable and fast method for subretinal injection in rodents and to assess the effect of the procedure on retinal function and histology. METHODS: Corneas of rodents were punctured with a 28 gauge hypodermic insulin needle avoiding the lens. The injection procedure can be observed with the aid of a dissecting microscope and methylcellulose solution on the eye. A 33 gauge blunt needle was inserted into the eye through the corneal puncture and guided toward the subretinal matrix. Addition of fluorescein to the injection mixture facilitated immediate evaluation of the injection. Rat eyes were either non-injected (controls), received only a corneal puncture or were injected with fluorescent microspheres or PBS-fluorescein mixture. Retinal function and integrity were assessed through electroretinographic (ERG) analysis and postmortem histology. RESULTS: The anterior injection procedure provided a fast and simple method for subretinal injections. In rats a successful subretinal delivery was achieved in more than 90%, with less than 5% of the injected eyes developing cataracts. No significant differences in b-wave ERG amplitudes in rodent eyes over a five-week period were observed between non-injected control eyes and subretinally injected eyes (1 to 10 microl of PBS-fluorescein or 2 microl fluorescent microspheres). Histological analysis revealed that re attachment of the rat retina occurred in approximately 1 day post-injection and the phagocytotic ability of RPE cells remained intact. CONCLUSIONS: This method was easily learned and required a minimum of equipment and animal preparation. With experience, 10 to 30 eyes could be injected per h. Furthermore, the injection procedure did not compromise the lens, retina or retinal pigment epithelium (RPE). PMID- 11436000 TI - PKCalpha and PKCgamma overexpression causes lentoid body formation in the N/N 1003A rabbit lens epithelial cell line. AB - PURPOSE: The overexpression of PKCalpha or PKCgamma for extended periods of time causes the formation of lentoid bodies in the N/N 1003A rabbit lens epithelial cell line. To determine how differentiated the lentoid bodies are, we have looked for alphaA-, alphaB-, beta-, and gamma-crystallin levels in lentoid bodies after 4 and 8 weeks of lentoid body development. METHODS: Cells overexpressing PKCalpha or PKCgamma were plated in 6 well plates and were allowed to form lentoid bodies for up to 8 weeks. Lentoid bodies were fixed and stained with PKCalpha or PKCgamma antibodies along with either alphaA-, alphaB-, beta-, or gamma crystallin antisera and viewed under a confocal microscope. Lentoid bodies were harvested in lysis buffer and homogenized. Fifty micrograms of protein per lane was loaded onto an SDS-PAGE gel and the bands transferred onto nitrocellulose. The blot was probed with either alphaA-, alphaB-, beta-, or gamma-crystallin antibodies for 12 h. Total RNA from lentoid bodies was isolated and 5 microg of total RNA was transcribed to first-strand cDNA. The PCR products were analyzed by 2% agarose gel electrophoresis. RESULTS: alphaB-crystallin was present in normal N/N 1003A cells and the lentoid bodies formed from PKCalpha and PKCgamma overexpression. alphaA-crystallin was only detectable in lentoid bodies after PKCalpha or PKCgamma overexpression. RT-PCR was able to detect beta-crystallin expression while the Western blot analysis and immunocytostaining detected small amounts of beta-crystallin protein. No gamma-crystallin expression was noted in these lentoid bodies. CONCLUSIONS: Overexpression of PKCalpha or PKCgamma in the N/N 1003A cell line induced lentoid body formation. These lentoid bodies expressed not only alphaB-crystallin but alphaA- and beta-crystallin. These results suggest a role for PKCs in lens epithelial cell differentiation to a fiber cell. PMID- 11436001 TI - Effect of H-7 on cultured human trabecular meshwork cells. AB - PURPOSE: To determine the effect of the serine-threonine kinase inhibitor H-7, which blocks actomyosin contractility and increases outflow facility in live monkeys, on morphology, cytoskeleton, and cellular adhesions of human trabecular meshwork (HTM) cells in culture. METHODS: Cultured HTM cells were videographically recorded and evaluated before and after exposure to H-7 at different concentrations. The subcellular distribution of the actin-based cytoskeleton and associated anchor proteins including vinculin, paxillin, and beta-catenin, as well as phosphotyrosine-containing proteins were evaluated by fluorescence immunocytochemistry and digital fluorescence microscopy. RESULTS: H 7 induced pronounced but reversible HTM cell thickening toward the cell center and deterioration of the actin cytoskeletal network. Cell-extracellular matrix (ECM) and cell-cell adhesions were also affected, but the beta-catenin-rich, vinculin-containing adherens junctions were clearly more resistant than focal contacts. Phosphotyrosine labeling in focal contacts was highly sensitive to H-7. CONCLUSIONS: H-7 induces alterations in cell shape, actin cytoskeleton, and associated focal adhesions in cultured HTM cells, which may be responsible for the effects of H-7 on outflow facility in live monkey eyes. PMID- 11436002 TI - Overview of medical devices. PMID- 11436003 TI - The investigation and management of the infant with ambiguous genitalia: a surgeon's perspective. PMID- 11436005 TI - Venous thromboembolism in trauma patients: a review. PMID- 11436006 TI - Helicobacter pylori and the gastroduodenal mucosa. PMID- 11436007 TI - Microsatellite instability, mismatch repair deficiency, and colorectal cancer. PMID- 11436008 TI - Postoperative and late survival outcomes after major amputation: findings from the Department of Veterans Affairs National Surgical Quality Improvement Program. AB - BACKGROUND: A surgical risk model is used to analyze postoperative mortality and late survival for older veterans who underwent above- or below-knee amputations in 119 Veterans Affairs (VA) hospitals from 1991 to 1995. METHODS: Preoperative medical conditions and laboratory values abstracted by the VA National Surgical Quality Improvement Program were linked to subsequent hospitalization and survival through 1999. Logistic regression and proportional hazards models were used to develop risk indexes for postoperative mortality and long-term survival. RESULTS: Thirty-day postoperative mortality was 6.3% for 1909 below-knee and 13.3% for 2152 above-knee amputees. Mortality varied greatly between the lowest highest risk index quartiles (0.8%-18.4% for below-knee amputation and 2.3%-31.1% for above-knee amputation). Surviving patients had 10,827 subsequent VA hospitalizations during a median 32-month follow-up. Survival probabilities for below- and above-knee amputees were 77% and 59% at 1 year, 57% and 39% at 3 years, and 28% and 20% at 7.5 years. The lowest quartile of survival risk had a 61% five-year survival compared with 14% for the highest-risk quartile. CONCLUSION: A generic surgical risk model can be of use in stratifying prognosis after major amputation. The heavy burden of hospital use by these patients suggests the need for better disease management for this high-risk, high-cost patient population. PMID- 11436009 TI - Conservative management of patients with intrathyroidal well-differentiated follicular thyroid carcinoma. AB - BACKGROUND: Total or near-total thyroidectomy for the treatment of follicular thyroid carcinoma (FTC). The prognosis of patients with low-risk FTC, however, is excellent, and thus total thyroidectomy may not be justifiable in such patients. METHODS: A retrospective review identified 61 patients diagnosed with intrathyroidal well-differentiated FTC between 1958 and 1991. RESULTS: Median age at diagnosis was 42 years (range, 15-78 years). Most patients (90.2%) had a lobectomy or subtotal thyroidectomy. Median tumor size was 3.0 cm (range, 0.9-9.5 cm). Fifty-eight patients (95.1%) received thyroid hormone supplementation, and 5 (8.2%) received radioactive iodine ablation postoperatively. Median follow-up was 11 years (range, 3-35 years). Local recurrence, metastasis, or both developed in 3 patients (4.9%), and all subsequently died of thyroid cancer. The cumulative 10 and 15-year cancer-specific survival rate was 96.5%. Factors significantly related to worse survival were oxyphilic histology (log-rank, P =.00) and tumor size of more than 4 cm (P =.001). However, neither was found to be an independent predictor of outcome by Cox multivariate analyses (P =.7 and.9, respectively). The extent of initial operation (unilateral versus bilateral procedure) was not significantly related to survival (P =.52). CONCLUSION: Conservative management consisting mainly of lobectomy or subtotal thyroidectomy and thyroid hormone supplementation is associated with favorable outcome of patients with intrathyroidal well-differentiated FTC. PMID- 11436010 TI - Appendectomy and subsequent risk of inflammatory bowel diseases. AB - BACKGROUND: Case-control studies have reported an inverse relationship between appendectomy and the risk of ulcerative colitis, but the association has not been confirmed in prospective studies. METHODS: Using national hospital discharge registry data in Denmark, the authors followed up 154,434 patients who underwent appendectomy during the period 1977 to 1989 to investigate whether they had subsequent hospitalizations for ulcerative colitis and Crohn's disease. Ratios of observed-to-expected first hospitalizations for inflammatory bowel diseases served as measures of the relative risk (RR). RESULTS: Hospitalization for ulcerative colitis occurred in 84 patients who had appendectomies versus 97.0 expected (RR = 0.87; 95% CI, 0.69-1.07). RRs were not significantly reduced in subgroups defined by sex, age, time since appendectomy, calendar period, or cause of appendectomy. Hospitalization for Crohn's disease occurred in excess (RR = 2.88; 95% CI, 2.45-3.39; n = 150), notably in the first year after appendectomy (RR = 10.83; 95% CI, 8.49-13.62; n = 73); but after 5 years, the RR was not significantly elevated. CONCLUSIONS: This large population-based cohort study failed to support a significant inverse association between appendectomy and ulcerative colitis risk in the first decade after the operation. The excess of Crohn's disease shortly after appendectomy most likely reflects differential diagnostic problems in patients newly presenting with abdominal pain. PMID- 11436011 TI - The induction of angiogenesis by the implantation of autologous bone marrow cells: a novel and simple therapeutic method. AB - BACKGROUND: Bone marrow contains many kinds of primitive cells that could differentiate to endothelial cells and secrete several growth factors. In the current study, we attempted to induce therapeutic angiogenesis by implanting autologous bone marrow cells (BMCs) and using a rat ischemic hind limb model. METHODS: BMCs were prepared by removing red blood cells. A rat ischemic hind limb model was made by the ligation of the left femoral artery and its branches. BMCs were injected into 7 points of the ischemic muscles. To assess angiogenesis, a microangiogram, laser Doppler, and histologic evaluation were performed after the surgical procedure. RESULTS: A microangiogram and histologic evaluation showed that angiogenesis was significantly induced in the ischemic hind limb by the implantation of BMCs. Laser Doppler imaging analysis showed that blood flow was significantly increased after implantation of BMCs. Some implanted BMCs were stained positively with CD31 and vascular endothelial-cadherin (VE-cadherin), which might have been incorporated into the vasculature. The condition of ischemia caused an elevation in the level of basic fibroblast growth factor in the ischemic muscle and also in interleukin-1beta derived from the implanted BMCs, which might contribute to angiogenesis. CONCLUSION: These findings indicate that autologous bone marrow implantation may be a novel and simple method for inducing therapeutic angiogenesis. PMID- 11436012 TI - Localized, benign, nontraumatic strictures of the extrahepatic biliary tree in children. AB - BACKGROUND: Benign, nontraumatic, inflammatory strictures of the extrahepatic biliary tree are rare in children and have been reported infrequently in the literature. We describe 7 children with this type of stricture and describe the results of their surgical treatment. METHODS: There were 6 girls and 1 boy, aged 2(1/2) to 15 years. The majority, who had no significant medical or surgical history, were first seen with obstructive jaundice. Investigations revealed isolated strictures of the extrahepatic biliary tree and varying degrees of secondary biliary change within the liver. All 7 patients underwent biliary enteric anastomosis; 5 also had resection of the stricture. RESULTS: No child experienced significant early complications from the operation, although 2 patients with unresectable lesions required further surgical treatment since their initial bypass. All patients are currently well at 1 to 17 years from initial referral without evidence of recurrent biliary disease after resection. CONCLUSIONS: Children who present with benign strictures of the extrahepatic biliary tree can be treated very satisfactorily with resection and hepaticojejunostomy. This rare condition should be considered as part of the differential diagnosis in children who present with obstructive jaundice. The etiology remains unknown. PMID- 11436013 TI - Efficacy of a hyaluronan derivative gel in postsurgical adhesion prevention in the presence of inadequate hemostasis. AB - BACKGROUND: We previously demonstrated that an auto-cross-linked hyaluronan-based antiadhesion agent (auto-cross-linked polysaccharide [ACP] gel) was effective in postsurgical adhesion prevention after open laparotomy and laparoscopic surgery with adequate hemostasis in animal models. This study assessed the ability of different preparations of ACP gel to prevent adhesions in the presence of bleeding or inadequate hemostasis. METHODS: Ninety-seven female rabbits were subjected to a standardized surgical lesion with subsequent exudative abdominal bleeding (oozing model), and 97 animals were subjected to a standardized surgical lesion with severe abdominal bleeding (bleeding model). After injury, the animals were randomly assigned to 5 groups of treatment: 3 different preparations of ACP gel (20, 40, and 60 mg/mL), a hyaluronan-carboxymethylcellulose film, and no treatment. Three weeks after operation, the animals were killed, and the adhesions were assessed by a blinded observer who measured the length and area of the adhesions and who used the Blauer scoring system. RESULTS: All 3 preparations of ACP gel and the hyaluronan-carboxymethylcellulose film reduced adhesion formation in both models (P <.01) as measured by the number of adhesion-free animals, mean Blauer score, and the mean length and surface area of the adhesions. There were no statistical differences between the different treatment groups. CONCLUSIONS: These data suggest that different hyaluronan based agents in the presence of severe bleeding or exudative abdominal bleeding reduce de-novo postsurgical adhesion formation. PMID- 11436014 TI - Surgical stress shifts the intestinal Escherichia coli population to that of a more adherent phenotype: role in barrier regulation. AB - BACKGROUND: We have shown that the combination of surgical stress and starvation in mice is associated with a defect in epithelial permeability and increased numbers of mucosa-associated Escherichia coli in the cecum. The aim of this study was to determine the specific role of mucosa-associated E coli on epithelial barrier dysfunction in this model. METHODS: Cecal E coli were harvested from mice 48 hours after a sham operation (control mice) or after a 30% surgical hepatectomy with only water provided ad libitum (short-term starvation) after the surgical procedure. Strains were tested for their ability to adhere to and alter the transepithelial electrical resistance (TEER) of cultured young adult mouse colon epithelial cells. TEER changes were further characterized by mannitol fluxes to confirm a defect in paracellular permeability. RESULTS: Strains of cecal E coli harvested from hepatectomy-starved mice adhered to and altered the permeability of young adult mouse colon cells, whereas E coli from the cecum of control mice were less adherent and had no effect on epithelial permeability. The effect of the strains harvested from mice after hepatectomy on the TEER of young adult mouse colon cells was inhibited by mannose and reversed by ciprofloxacin. CONCLUSION: The combination of surgical stress and short-term starvation is associated with a greater abundance of adherent and barrier-altering strains of E coli in the mouse cecum. PMID- 11436015 TI - Selective versus total drainage for biliary obstruction in the hepatic hilus: an experimental study. AB - BACKGROUND: Controversy exists as to whether selective biliary drainage (SBD) or total biliary drainage (TBD) is preferable as a preoperative procedure before extended hemihepatectomy for hilar cholangiocarcinoma, especially with regard to the functional reserve of the future remnant liver. METHODS: SBD or TBD was performed after 1 week of total biliary obstruction in rats. In SBD, the biliary trees of the left lobes (approximately 70% of the liver) were kept obstructed, whereas the right lobes were drained selectively. Mitochondrial function and microsomal cytochrome content were examined before and 1, 2, and 4 weeks after drainage. RESULTS: The right lobes weighed significantly more after SBD than after TBD. There were no significant differences in mitochondrial function between the two groups. The microsomal cytochrome content per milligram of microsomal protein significantly decreased 1 week after biliary obstruction and then recovered to a similar extent after SBD and TBD in the right lobes. However, the total microsomal cytochrome content (nanomoles per 100 g body weight) and the overall rate of mitochondrial adenosine triphosphate synthesis (mmoles per minute per 100 g of body weight) in the right lobes 4 weeks after SBD were significantly greater than those after TBD. CONCLUSIONS: SBD is superior to TBD with regard to the functional reserve of the future remnant liver. PMID- 11436016 TI - Circumstance and serendipity. PMID- 11436017 TI - Duodenal duplication in a young adult. PMID- 11436018 TI - Preoperative diagnosis of acute appendiceal diverticulitis by ultrasonography. PMID- 11436019 TI - Slow progression and sequential documentation of a giant basal cell carcinoma of the face. PMID- 11436020 TI - Electromyographic nerve identification during resection of an intrathoracic goiter via a right anterolateral thoracotomy using a novel method. PMID- 11436021 TI - Edge deepithelialization: a method to prevent leakage when tubed free skin flap is used for pharyngoesophageal reconstruction. AB - BACKGROUND: Anastomosis leakage remains a major source of morbidity and mortality in patients who receive pharyngoesophageal reconstruction, especially when a free skin flap is used as the substitute. This makes free skin flaps less favored than other substitutes in pharyngoesophageal reconstruction. However, a free skin flap provides many advantages for pharyngoesophageal reconstruction and, in some circumstances, may be the only choice. To reduce the incidence of postoperative leakage, we developed a technical modification called edge-deepithelialization for skin flap to ensure a more secure anastomosis. METHODS: Twenty-four patients who received tubed free skin flaps for cervical esophagus or hypopharynx reconstruction during a 7-year period were included in this study. Group 1 (n = 12) received flap edge-deepithelialization before undergoing the anastomosis, and Group 2 (n = 12) received the conventional anastomosis method. Preoperative nutritional status was evaluated by serum albumin level. All the patients were followed up for at least 1 year (1 approximately 14 years; mean, 7.3 years). Postoperative leakage was detected by clinical symptoms and signs or by routine esophagography. RESULTS: Postoperative anastomosis leakage developed in 1 patient in Group 1 (8.3%) and 5 patients in Group 2 (41.67%). The difference in the leakage rate between these groups was of statistical significance (P =.0001). There were no significant differences in the preoperative serum albumin level noted between these 2 groups (t test, P >.05). Older age was not associated with a higher leakage rate. Among the 6 leakage cases, 1 was treated conservatively, and the other 5 needed an additional 1 to 5 procedures for the management of leakage. All the leakages were cured during the follow-up period. CONCLUSIONS: With the flap edge deepithelialized, a more secure and reliable anastomosis can be obtained. This makes the free skin flap an easier and safer alternative in pharyngoesophageal reconstruction, especially when laparotomy and enteric flaps are contraindicated. PMID- 11436022 TI - Eosinophilic abscess and eosinophilic pseudotumor presenting as bile duct masses: a report of 2 cases. PMID- 11436023 TI - Retroperitoneal follicular dendritic cell sarcoma presenting as secondary amyloidosis. PMID- 11436024 TI - Duodenal duplication cysts: A rare cause of acute pancreatitis in children. PMID- 11436025 TI - Portal embolization relieves persistent jaundice after complete biliary drainage. PMID- 11436026 TI - Invited commentary: Minimally invasive follicular thyroid carcinoma. PMID- 11436027 TI - Are appendectomy and the subsequent risk of ulcerative colitis related? PMID- 11436029 TI - Bulk buying. PMID- 11436030 TI - In search of the ideal valve replacement device. PMID- 11436031 TI - Outcome of bilateral lung volume reduction in patients with emphysema potentially eligible for lung transplantation. AB - OBJECTIVE: Between January 1993 and May 1998, we performed 200 consecutive bilateral lung volume reduction operations. After initial assessment, 99 of these patients were eligible for lung volume reduction and potentially eligible for immediate or eventual lung transplantation on the basis of age and absence of contraindications. All chose to proceed with lung volume reduction surgery. The outcomes of these 99 patients are reviewed to assess the consequences of proceeding with lung volume reduction surgery on patients potentially eligible for lung transplantation. METHODS: A retrospective study was performed with the use of a prospectively assembled computer database. RESULTS: The 61 men and 38 women were 55 +/- 7 years old at evaluation for lung volume reduction. Mean values for first second expired volume, total lung capacity, and residual volume were 24% +/- 8%, 141% +/- 19%, and 294% +/- 54% predicted. There were 4 operative deaths and 17 late deaths. Two-year and 5-year survival after evaluation for lung volume reduction are 92% and 75%. The 32 patients who have been listed for transplantation after lung volume reduction include 15 who have undergone transplantation, 14 who remain on the list, and 3 who have been removed from the list. All 15 transplant recipients survived transplantation and 3 have subsequently died of rejection or late infection. The 12 living recipients have a median post-transplantation follow-up of 1.7 years. The age at transplantation was 58 +/- 5 years with transplantation occurring 3.8 +/- 1.1 years after lung volume reduction. Sixteen of 99 patients underwent lower lobe volume reduction with an increased rate of listing (63%, P =.008) and transplantation (38%, P =.003) compared with patients undergoing upper lobe volume reduction. Patients listed for transplantation were younger, more impaired, and experienced less benefit from lung volume reduction than patients not yet listed for transplantation. CONCLUSIONS: The preliminary use of lung volume reduction in patients potentially suitable for transplantation does not appear to jeopardize the chances for subsequent successful transplantation. PMID- 11436032 TI - Lung retrieval from non-heart beating cadavers with the use of a rat lung transplant model. AB - BACKGROUND: Lungs retrieved from cadavers after death and circulatory arrest may alleviate the critical shortage of lungs for transplant. We report a rat lung transplantation model that allows serial measurement of arterial blood gases after left single lung transplantation from non-heart beating donors. METHODS: Twelve Sprague-Dawley rats underwent left lung transplantation with a vascular cuff technique. Donor rats were anesthetized with intraperitoneal injection of pentobarbital, heparinized, intubated via tracheotomy, and then killed with pentobarbital. Lungs were retrieved immediately or after 2 hours of oxygen ventilation after death (tidal volume 1 mL/100 g, rate 40/min FIO2 = 1.0, positive end-expiratory pressure 5 cm H2O). Recipient rats were anesthetized, intubated, and ventilated. The carotid artery and jugular vein were cannulated for arterial blood gases and infusion of Ringer's lactate (4 mL/h). Anesthesia was maintained with halothane 0.2%, and recipient arterial blood gases were measured at 4 and 6 hours after lung transplantation after snaring the right pulmonary artery for 5 minutes. Animals were put to death 6 hours after lung transplantation, and portions of transplanted lungs were frozen in liquid nitrogen and assayed for wet/dry ratio, myeloperoxidase as a measure of neutrophil infiltration, and conjugated dienes as a measure of free radical mediated lipid peroxidation. RESULTS: Arterial PO2 and wet/dry ratio were not significantly different in recipients of non-heart beating donor lungs retrieved immediately after death or after 2 hours of oxygen ventilation. Significant neutrophil infiltration was observed in recipients of non-heart beating donor lungs retrieved 2 hours after death from oxygen-ventilated donors. CONCLUSIONS: Strategies to ameliorate reperfusion injury may allow for successful lung transplantation from non-heart beating donors. PMID- 11436033 TI - Intrapulmonary metastasis of non-small cell lung cancer: a prognostic assessment. AB - OBJECTIVE: According to the revised TNM classification in 1997, intrapulmonary metastasis within the same lobe of the primary tumor is designated as T4 and intrapulmonary metastasis in a different lobe is M1. However, their prognostic implications remain unclear. To assess their prognoses, we retrospectively analyzed the postoperative survival of patients with and without intrapulmonary metastasis. METHODS: From January 1982 to December 1996, 2340 patients with non small cell lung cancer underwent surgical resection. The survival of patients having complete resection (n = 1534) was analyzed according to their intrapulmonary metastasis status: patients without intrapulmonary metastasis (n = 1393), those with metastasis in the same lobe (n = 105), and those with metastasis in a different lobe (n = 18). For comparison, patients with T4 disease without intrapulmonary metastasis in the same lobe (n = 54) and those with M1 disease without metastasis in a different lobe (distant M1, n = 18) were also analyzed. RESULTS: The overall 5-year survivals were as follows: no intrapulmonary metastasis, 60%; stage T4 disease with no intrapulmonary metastasis, 34%; pulmonary metastasis in the same lobe, 34%; pulmonary metastasis in a different lobe, 11%; and distant M1, 6%. The differences in survival between patients with no pulmonary metastasis and those with metastasis in the same lobe (P <.001, log-rank test) and between patients with metastasis in the same lobe and those with distant M1 (P <.001) were significant. In contrast, there was no significant difference between patients with metastasis in the same lobe and those with T4 disease and no intrapulmonary metastasis or between patients with metastasis to a different lobe and those with distant M1. CONCLUSIONS: Prognostically, intrapulmonary metastasis within the same lobe of the primary tumor was comparable with T4 and that in a different lobe was comparable with M1. In terms of postoperative prognosis, the revised TNM classification for intrapulmonary metastasis seems to be appropriate. PMID- 11436034 TI - Lipid-mediated ex vivo gene transfer of viral interleukin 10 in rat lung allotransplantation. AB - BACKGROUND: Recent studies suggest that viral interleukin 10 suppresses alloimmune response in transplantation and that cationic lipids are one of the most promising nonviral vehicles for gene therapy. The aim of this study was to examine the effect of ex vivo lipid-mediated viral IL10 gene transfer into rat lung allografts on subsequent rejection. METHODS: Male F344 rats (RT1lvl) underwent left lung transplantation with allografts from Brown Norway rats (RT1n). Allografts were transvascularly transfected 15 minutes after harvest with 5 mL of 1:20-diluted (group 1, n = 7) or 1:40-diluted (group 2, n = 6) GL67 pCMVievIL-10 complex. Group 3 (n = 7), serving as the control group, received 1:40-diluted GL67-pCF1-chloramphenicol acetyltransferase complex. All allografts were preserved for 3 hours at 10 degrees C before transplantation. In all groups recipients were killed on postoperative day 5. Transgene expression of viral interleukin 10 was assessed by means of both reverse transcriptase-polymerase chain reaction and immunohistochemistry. Histologic rejection score, allograft gas exchange, exhaled nitric oxide level, and allograft cytokine mRNA expression were also assessed. RESULTS: Dose-dependent transgene expression of viral interleukin 10 was detected by means of both reverse transcriptase-polymerase chain reaction and immunohistochemistry. Allograft gas exchange (PaO2) in groups 1 (114.06 +/- 61.1 mm Hg) and 2 (108.58 +/- 35.7 mm Hg) was significantly better than that in group 3 (66.4 +/- 8.22 mm Hg; P =.020 and P =.023, respectively). The vascular rejection score in group 1 was significantly lower than that in group 3 (P =.032, Kruskal-Wallis test). Exhaled nitric oxide levels in group 2 (5.150 +/- 6.38 ppb) were significantly lower than those in group 3 (13.517 +/- 10.4 ppb; P =.039). Allograft interleukin 2 mRNA expression levels in group 1 (1.123 +/- 0.23 relative units) were significantly lower than those in group 3 (1.753 +/- 0.71 relative units; P =.038 vs group 3). CONCLUSIONS: Lipid-mediated ex vivo viral IL10 gene transfer into rat lung allografts improved graft gas exchange, reduced histologic rejection scores, downregulated graft interleukin 2 mRNA expression, and reduced exhaled nitric oxide levels by postoperative day 5. These results suggest a therapeutic potential of graft viral IL10 gene transfer as an effective immunosuppressive strategy against lung allograft rejection. PMID- 11436035 TI - Results of aortic valve-sparing operations. AB - OBJECTIVE: To review the late results of valve-sparing operations in patients with aortic root aneurysm and in those with ascending aortic aneurysm and aortic insufficiency. METHODS: From May 1988 to June 2000, 120 patients with aortic root aneurysm and 68 with ascending aortic aneurysm and aortic insufficiency underwent aortic valve-sparing operations. Patients with aortic root aneurysm were younger, were predominantly male, and had less severe aortic insufficiency than patients with ascending aortic aneurysm, who were older and often had aneurysm of the transverse arch. Forty-eight patients with aortic root aneurysm had the Marfan syndrome. The prevalence of aortic dissection was similar in both groups. Reconstruction of the aortic root was performed by reimplanation of the aortic valve in 64 patients and by remodeling of the aortic root in 56. Patients with ascending aortic aneurysm and aortic insufficiency were treated by replacement of the ascending aorta with reduction in the diameter of the sinotubular junction. Approximately two thirds of the latter patients also required replacement of the transverse aortic arch. The mean follow-up was 35 +/- 31 months for patients with aortic root aneurysm and 26 +/- 23 months for those with ascending aortic aneurysm. RESULTS: There were 2 operative and 5 late deaths in patients with aortic root aneurysm and 1 operative and 9 late deaths in patients with ascending aortic aneurysm. The 5-year survival for patients with aortic root aneurysm was 88% +/- 4% and for patients with ascending aortic aneurysm, 68% +/- 12% (P =.01). Severe aortic insufficiency developed in 2 patients, and they required aortic valve reoperation. The 5-year freedom from aortic valve reoperation was 99% +/- 1% for patients with aortic root aneurysm and 97% +/- 4% for those with ascending aortic aneurysm. Seven patients had moderate aortic insufficiency at the latest echocardiographic study. The 5-year freedom from severe or moderate aortic insufficiency was 90% +/- 4% in patients who had aortic root aneurysm and 98% +/- 2% in those who had ascending aortic aneurysm. CONCLUSIONS: Aortic valve-sparing operations have provided excellent clinical outcomes and few valve-related complications. The function of the reconstructed aortic root remains unchanged in most patients during the first 5 years of follow-up. PMID- 11436036 TI - Changes in aneurysm morphology and stent-graft configuration after endovascular repair of aneurysms of the descending thoracic aorta. AB - OBJECTIVE: We sought to study changes in morphology and stent-graft configuration of descending thoracic aortic aneurysms after endovascular repair. METHODS: Twenty-three patients treated with custom-made stent-grafts were studied. The stent-graft consisted of continuous, stainless-steel Z stents mounted within a polyester graft. In the last 11 cases the stents were interconnected with 3 longitudinal wires. Contrast-enhanced spiral computed tomography was performed preoperatively and at 1, 3, and every 6 months postoperatively. Angiography was used preoperatively and at 1-year follow-up. Proximal and distal necks were assessed for diameter and length. Aneurysm diameter, endoleaks, stent-graft migration, and changes in stent-graft configuration were evaluated. RESULTS: During follow-up (median, 18 months; range, 1-48 months), excluded aneurysms decreased in diameter by 4 mm (0.5-10 mm, P =.0018). Endoleaks prevented size decrease. Five patients displayed neck dilatation, 4 at both the proximal and distal fixation sites and 1 only distally. In 7 (30%) patients there was proximal migration of the distal end of the stent-graft. Three (13%) patients displayed both distal migration of the proximal end of the stent-graft and proximal migration of the distal end of the stent-graft. There was a significant correlation between stent-graft kinking and appearance of proximal or distal stent-graft migration (P =.05 and P =.0007, respectively). In no case did the migration lead to appearance of an endoleak before intervention was performed. CONCLUSION: Excluded descending thoracic aortic aneurysms decrease in size on midterm follow-up. A subgroup of patients prone to neck dilatation might exist. A combination of neck dilatation and vector forces acting on stent-grafts in the tortuous thoracic aorta might lead to stent-graft migration. PMID- 11436037 TI - Relationship of hospital size, case volume, and cost for coronary artery bypass surgery: analysis of 12,774 patients operated on in Massachusetts during fiscal years 1995 and 1996. AB - OBJECTIVE: This study investigates the relationship between the cost of coronary artery bypass graft surgery and both hospital size and case volume. METHODS: Retrospective administrative and cost data were obtained for all 12,774 patients who underwent isolated coronary bypass surgery at 12 Massachusetts hospitals during 1995 and 1996. Hospitals were stratified by number of operating beds into 3 groups (group I, <250 beds; group II, 250-450 beds; group III, >450 beds). Total (diagnosis-related groups 106 + 107) annual coronary bypass cases per hospital varied from 271 to 913 (mean 532). Univariate and multivariable analyses were used to study the relationship between the direct and total cost and a number of patient (age, sex, acuity class, payer) and hospital (bed capacity, annual case volume per diagnosis-related group, cardiothoracic residency) predictor variables. For each hospital, we also studied the relationship between changes in coronary bypass case volume and the corresponding changes in average cost from 1995 to 1996. RESULTS: Scatterplots revealed a broad range of mean direct cost of coronary bypass surgery among hospitals with comparable case volumes. When annual cases were analyzed as continuous variables, there was no linear relationship of case volume with direct or total cost of coronary bypass (r = -0.05 to +0.08) for any diagnosis-related group or year. When hospital bed capacity and case volume were grouped into strata and studied by analysis of variance, there was no evidence of an inverse relationship between these variables and cost. In multivariable analysis, patient acuity class and diagnosis related group were the most important predictors of cost. Beds and case volume met inclusion criteria for most models but added little to the "explanation" of variability R(2), often less than 1%. Finally, substantial interhospital differences were noted in the magnitude and direction (direct vs inverse) of their 1995 to 1996 change in volume versus change in cost. CONCLUSIONS: Within the range of hospital size and case volume represented in this study, there is no evidence that either variable is related to the cost of performing coronary bypass surgery. Massachusetts hospitals appear to function on different segments of different average cost curves. It is not possible to predict the relative cost of coronary bypass grafting at a given hospital based primarily on volume. PMID- 11436038 TI - Human angiopoietin gene expression is a marker for severity of pulmonary hypertension in patients undergoing pulmonary thromboendarterectomy. AB - OBJECTIVE: A consistent pathologic feature seen in lungs of patients with pulmonary hypertension from thromboembolic disease is hyperplasia of the media of pulmonary arterioles. The molecular factors responsible for these vessel wall changes are unknown. Angiopoietin-1 is a gene responsible for the formation of the media of blood vessels in utero. We hypothesized that aberrant expression of the angiopoietin-1 gene in the adult lung would be a major contributing factor in the development of pulmonary hypertension. METHODS: From April 1999 to March 2000, a total of 35 patients (18 men, 17 women, mean age 52 years) with pulmonary hypertension and pulmonary vascular resistance ranging from 407 to 2006 dynes x sec x cm(-5) underwent pulmonary endarterectomy at our institution. Before cardiopulmonary bypass, lung biopsy specimens were taken from each patient. Biopsy specimens were also obtained from 10 patients (5 women, 5 men, mean age 55 years) undergoing lung resection for causes other than pulmonary hypertension. All specimens were blindly scored by a pathologist for degree of medial hyperplasia. Quantitative reverse transcriptase-polymerase chain reaction, Western blot, and immunohistochemistry were used to quantitate angiopoietin-1 messenger RNA and protein in each sample. RESULTS: Lung specimens from all patients with pulmonary hypertension demonstrated up-regulation of angiopoietin-1 at the messenger RNA level. The degree of angiopoietin-1 transcription was directly proportional to the preoperative pulmonary vascular resistance and medial wall hyperplasia/hypertrophy in each patient. By immunohistochemistry, angiopoietin-1 protein was confined to the media of pulmonary arterioles. Lung biopsy specimens from patients without pulmonary hypertension had no detectable expression of angiopoietin-1 at the messenger RNA or protein level. CONCLUSION: Angiopoietin-1, a gene responsible for vessel development in the embryonic lung, is up-regulated in the lung parenchyma of patients with pulmonary hypertension. The level of expression of angiopoietin-1 at messenger RNA and protein levels correlates to the severity of pulmonary hypertension in patients with thromboembolic disease and serves as a target for strategies to treat this disease. PMID- 11436039 TI - Bicuspid aortic valve disease and pulmonary autograft root dilatation after the Ross procedure: a clinicopathologic study. AB - OBJECTIVE: Bicuspid aortic valve disease has been associated with histologic abnormalities of the aortic root. Recent reports have suggested similar alterations may exist in the pulmonary artery of patients with bicuspid aortic valve. The present study was undertaken to define the histologic condition of the aortic and pulmonary artery root in bicuspid aortic valve disease and the relationship with pulmonary autograft root dilatation after the Ross procedure. METHODS: In 17 patients undergoing aortic root replacement with the pulmonary autograft, biopsy specimens of the aortic root and pulmonary artery trunk were collected. Clinical and histologic findings of patients with bicuspid aortic valves were compared with those with tricuspid aortic valves. RESULTS: There were 9 patients (8 male, 1 female) with bicuspid aortic valve (group 1) and 8 (all male) with tricuspid aortic valve (group 2). Mean age was comparable (24.4 +/- 9.8 vs 23.6 +/- 10.8 years, P =.9). Aortic insufficiency as an indication for operation was more common in group 1 (9/9 vs 5/8, P =.007), whereas preoperative aortic root dilatation was equally prevalent (4/9 vs 1/8, P =.1). Prior aortic valve repair had been performed in 2 patients (1/9 vs 1/8, P =.9). Prevalence of cystic medionecrosis of the aortic wall was similar in the 2 groups (4/9 vs 3/8, P =.6). Cystic medionecrosis of the pulmonary artery trunk was found only in 1 patient with tricuspid aortic valve (0/9 vs 1/8, P =.3). During a mean follow-up of 26.5 +/- 12.2 months (32.1 +/- 12.7 vs 20.1 +/- 7.4 months, P =.04), prevalence of pulmonary autograft root dilatation (greater than 4.0 cm) was equally represented in patients with native bicuspid or tricuspid aortic valve (3/9 vs 2/8, P =.6). CONCLUSIONS: Histologic abnormalities of the pulmonary artery root are rare and equally prevalent in young patients with bicuspid and tricuspid aortic valves. On the contrary, root dilatation is relatively common late after autograft root replacement but appears unrelated to bicuspid aortic valve disease or to pre-existing degenerative changes of the pulmonary artery root. PMID- 11436040 TI - Modified Glenn connection for acutely ischemic right ventricular failure reverses secondary left ventricular dysfunction. AB - BACKGROUND: Right heart failure after cardiopulmonary bypass can result in severe hemodynamic compromise with high mortality, but the underlying mechanisms remain poorly understood. After ischemia-induced right ventricular failure, alterations in the interventricular septal position decrease left ventricular compliance and limit filling but may also distort left ventricular geometry and compromise contractility and relaxation. This study investigated the effect of acute isolated right ventricular ischemia on biventricular performance and interaction and the response of subsequent right ventricular unloading by use of a modified Glenn shunt. METHODS: In 8 pigs isolated right ventricular ischemic failure was induced by means of selective coronary ligation. A modified Glenn circuit was then established by a superior vena cava-pulmonary artery connection. Ventricular performance was determined by conductance catheter-derived right ventricular pressure-volume loops and left ventricular pressure-segment length loops. Hemodynamic data at baseline, after right ventricular ischemia, and after institution of the Glenn circuit were obtained during inflow occlusion, and the load-independent contractile indices were derived. RESULTS: Right ventricular free-wall ischemia resulted in acute right ventricular dilation (118 +/- 81 mL vs 169 +/- 70 mL, P =.0008) and impairment of left ventricular contractility indicated by the reduced end-systolic pressure-volume relation slope (50.0 +/- 19 mm Hg/mm vs 18.9 +/- 8 mm Hg/mm, P =.002) and preload recruitable stroke work index slope (69.6 +/- 26 erg x cm(-3) x 10(3) vs 39.7 +/- 13 erg x cm(-3) x 10(3), P =.003). In addition, left ventricular relaxation (tau) was significantly prolonged (33.3 +/- 10 ms vs 53.0 +/- 16 ms, P =.012). Right ventricular unloading with the Glenn shunt reduced right ventricular dilation and significantly improved left ventricular contraction, end-systolic pressure-volume relation slope (18.9 +/- 8 mm Hg/mm vs 35.8 +/- 18 mm Hg/mm, P =.002), preload recruitable stroke work index slope (39.7 +/- 26 erg x cm(-3) x 10(3) vs 63.0 +/- 22 erg x cm(-3) x 10(3), P =.003), and diastolic performance (tau 53.0 +/- 16 ms vs 43.5 +/- 13 ms, P =.001). CONCLUSIONS: Right ventricular ischemia-induced dilation resulted in acute impairment of left ventricular contractility and relaxation. A modified Glenn shunt attenuated the left ventricular dysfunction by limiting right ventricular dilation and restoring left ventricular cavity geometry. PMID- 11436041 TI - Clinical experience with 202 adults receiving extracorporeal membrane oxygenation for cardiac failure: survival at five years. AB - OBJECTIVES: We sought to determine 5-year survival after extracorporeal membrane oxygenation for cardiac failure and its predictors, to assess survival and its predictors after bridging to transplantation or weaning from extracorporeal membrane oxygenation, and to identify factors influencing the likelihood of these outcomes. METHODS: Two hundred two adults (mean age, 55 +/- 14 years) were supported with extracorporeal membrane oxygenation between 1992 and July 1999 after cardiac failure. Follow-up extended to 7.5 years (mean, 3.8 +/- 2 years). Multivariable hazard function analysis identified predictors of survival, and logistic regression identified the determinants of bridging or weaning. RESULTS: Survival at 3 days, 30 days, and 5 years was 76%, 38%, and 24%, respectively. Patients surviving 30 days had a 63% 5-year survival. Risk factors (P <.1) included older age, reoperation, and thoracic aorta repair. Forty-eight patients were bridged to transplantation, and 71 were weaned with intent for survival. Survival was similar after either outcome (44% vs 40% 5-year survival, respectively). Failure to bridge or wean included (P <.03) renal and hepatic failure on extracorporeal membrane oxygenator support, occurrence of a neurologic event, and absence of infection. The dominant modes of death were cardiac failure and multisystem organ failure. CONCLUSIONS: Extracorporeal membrane oxygenation is versatile and salvages some patients who would otherwise die. Improvement in intermediate-term outcome will require a multidisciplinary approach to protect organ function and limit organ injury before and during this support. PMID- 11436042 TI - alpha1-Adrenoceptors during simulated ischemia and reoxygenation of the human myocardium: effect of the dose and time of administration. AB - OBJECTIVE: We sought to investigate the effect of alpha1-adrenoceptor activity on the ischemic and reoxygenated human myocardium. METHODS: Right atrial appendages (n = 6 per group) obtained during elective cardiac operations were sliced and stabilized in normoxic normothermic buffer solution for 30 minutes and then subjected to 90 minutes of simulated ischemia, followed by 120 minutes of reoxygenation. In study 1 the dose responses to the alpha1-adrenoceptor agonist phenylephrine (0.01, 0.1, 1, 10, and 100 micromol/L) and to the alpha1 adrenoceptor antagonist prazosin (0.1, 1, 10, and 100 micromol/L) when administered for 10 minutes before ischemia, during ischemia, and during reoxygenation were examined. The influence of the time of administration (ie, before ischemia, during ischemia, or during reoxygenation) of phenylephrine (0.1 micromol/L) and prazosin (10 micromol/L) was then investigated in study 2. In study 3 the effect of the combined administration of phenylephrine given before ischemia and prazosin given during ischemia was investigated. In study 4 the protective effect of phenylephrine given before ischemia (for 10 minutes or for 5 minutes with a 5-minute washout period) was compared with that of ischemic preconditioning (5 minutes of ischemia and 5 minutes of reoxygenation). At the end of each protocol, the leakage of creatine kinase (in units per gram of wet weight) and the reduction of 3-[4,5 dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide to insoluble formazan dye (in millimoles per gram of wet weight) were measured. RESULTS: Phenylephrine is maximally beneficial at 0.1 and 1 micromol/L (creatinine kinase, 0.97 +/- 0.06 and 0.95 +/- 0.03 U/g, respectively; 3-[4,5 dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide, 153.0 +/- 7.8 and 156.2 +/ 6.7 mmol/g, respectively) compared with ischemic control (creatine kinase, 1.87 +/- 0.03 U/g; 3-[4,5 dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide, 108.5 +/- 6.8 mmol/g; P <.05) but prazosin is detrimental at concentrations above 10 micromol/L (creatine kinase, 5.22 +/- 0.29 U/g; 3-[4,5 dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide, 69.8 +/- 2.9 mmol/g; P <.05 vs ischemic control). In addition, phenylephrine (0.1 micromol/L) is protective when given before ischemia (creatine kinase, 2.06 +/- 0.21 U/g; 3-[4,5 dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide, 148.5 +/- 4.5 mmol/g; P <.05 vs ischemic control) but is detrimental when given during ischemia alone (creatine kinase, 4.49 +/- 0.98 U/g; 3-[4,5 dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide, 70.5 +/- 6.1 mmol/g; P <.05 vs ischemic control) and has no significant effect during reoxygenation. In contrast, prazosin (10 micromol/L) is beneficial when given during ischemia alone (creatine kinase, 1.34 +/- 0.10 U/g; 3-[4,5 dimethylthiazol 2-yl]-2,5 diphenyltetrazolium bromide, 148.5 +/- 4.5 mmol/g; P <.05 vs ischemic control), is detrimental when given during reoxygenation alone (creatine kinase, 1.5 +/- 0.16 U/g; 3-[4,5 dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide, 85.0 +/- 4.7 mmol/g; P <.05 vs ischemic control), and has no effect when given before ischemia. The use of phenylephrine before ischemia alone is as protective as prazosin given during ischemia alone, but the combination of the two drugs does not cause additional benefit. Interestingly, the protection afforded by phenylephrine when given before ischemia is similar to that obtained with ischemic preconditioning. CONCLUSIONS: In the human myocardium activation of alpha1-adrenoceptors before ischemia is protective but is detrimental during ischemia, whereas blockade of alpha1-adrenoceptors is beneficial during ischemia but detrimental during reoxygenation. The degree of protection achieved by activation of the alpha1-adrenoceptors before ischemia is similar to that obtained with blockade of alpha1-adrenoceptors during ischemia and that of ischemic preconditioning. PMID- 11436043 TI - Inhibition of complement, neutrophil, and platelet activation by an anti-factor D monoclonal antibody in simulated cardiopulmonary bypass circuits. AB - OBJECTIVES: Patients undergoing cardiopulmonary bypass frequently manifest generalized systemic inflammation and occasionally manifest serious multiorgan failure. Inflammatory responses of bypass are triggered by contact of blood with artificial surfaces of the bypass circuits, surgical trauma, and ischemia reperfusion injury. We studied the effects of specific inhibition of the alternative complement cascade by using an anti-factor D monoclonal antibody (166 32) in extracorporeal circulation of human whole blood used as a simulated model of cardiopulmonary bypass. METHODS: Five healthy blood donors were used in the study. Monoclonal antibody 166-32 was added to freshly collected, heparinized human blood recirculated in a pediatric cardiopulmonary bypass circuit at a final concentration of 18 microg/mL. An irrelevant monoclonal antibody was used as a negative control with the same donor blood in a parallel bypass circuit on the same day. Blood samples were collected at different time points during recirculation for measurement of activation of complement, neutrophils, and platelets by immunofluorocytometric methods and enzyme-linked immunosorbent assays. RESULTS: Monoclonal antibody 166-32 inhibited the alternative complement activation and the production of Bb, C3a, sC5b-9, and C5a. Upregulation of CD11b on neutrophils and CD62P on platelets was also significantly inhibited by monoclonal antibody 166-32. This is consistent with the inhibition of the release of neutrophil-specific myeloperoxidase and elastase and platelet thrombospondin. The production of proinflammatory cytokine interleukin 8 was also suppressed by the antibody. CONCLUSIONS: The alternative complement cascade is predominantly activated during extracorporeal circulation. Anti-factor D monoclonal antibody 166-32 is effective in inhibiting the activation of complement, neutrophils, and platelets. Inhibition of the alternative complement pathway by targeting factor D could be useful in reducing systemic inflammation in patients undergoing cardiopulmonary bypass. PMID- 11436044 TI - Effect of aprotinin on endothelial cell activation. AB - BACKGROUND: Cardiopulmonary bypass surgery is often accompanied by a systemic inflammatory response, which can lead to postoperative complications in high-risk patients. This is mediated in part through a systemic rise in inflammatory cytokine levels and the sequestration of leukocytes within organs. Aprotinin has previously been shown to exert an anti-inflammatory effect by preventing the capacity of leukocytes to transmigrate through vascular endothelium. Here we have focused on whether aprotinin has an effect on endothelial cell activation and adhesion molecule expression in response to tumor necrosis factor-alpha, particularly with reference to whether aprotinin inhibits tumor necrosis factor stimulated neutrophil transendothelial migration. METHODS AND RESULTS: Intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and E selectin expression was studied in tumor necrosis factor-alpha-activated human umbilical vein endothelial cells in the presence of aprotinin at 200, 800, and 1600 kIU/mL. Aprotinin inhibited tumor necrosis factor-alpha-stimulated expression of intercellular adhesion molecule-1 (P =.019 at 1600 kIU/mL) and vascular cell adhesion molecule-1 (P =.003 at 1600 kIU/mL) but not E-selectin. Similar results were obtained in the dermal microvascular endothelial cell line, HMEC-1, which exhibited diminished intercellular adhesion molecule-1 expression in the presence of aprotinin (P =.040 at 800 kIU/mL and P <.001 at 1600 kIU/mL). Aprotinin also significantly inhibited neutrophil transmigration across tumor necrosis factor-alpha-activated human umbilical vein endothelial cells (P =.046 at 1600 kIU/mL). CONCLUSIONS: We have demonstrated that aprotinin inhibits intercellular adhesion molecule-1 and vascular cell adhesion molecule-1, but not E-selectin, expression on tumor necrosis factor-alpha-activated endothelial cells and that transendothelial migration by neutrophils is also specifically suppressed under these conditions. Our results indicate that endothelial cells can be specifically targeted by aprotinin, therefore adding to our understanding of the anti-inflammatory mechanism of action of aprotinin during cardiopulmonary bypass. PMID- 11436045 TI - Valve interstitial cells induce donor-specific T-cell anergy. AB - OBJECTIVES: Valve allografts produce an immune response, which can influence their performance. The exact role of the interaction between recipient T cells and the different cellular components of the donor valve in stimulating an immune response is not known. Therefore the T-cell response to valve endothelial and interstitial cells was investigated in vitro. METHODS: Valve endothelial and interstitial cells were characterized for cell-surface molecules before and after interferon gamma treatment by means of a panel of specific monoclonal antibodies and flow cytometry. The proliferative response of highly purified T lymphocytes was used to assess the immunogenicity of cultured valve endothelial and interstitial cells. This was further investigated by using a 2-step tolerance induction protocol. RESULTS: Valve endothelial and interstitial cells express similar levels of human leukocyte antigens and adhesion and costimulatory molecules, which are either induced or upregulated after interferon gamma treatment. T-cell responses to endothelial cells were detected after interferon gamma treatment, but responses to interferon gamma-treated interstitial cells were not detected. This lack of response resulted in the induction of T-cell anergy, which was reversed by the presence of the costimulatory molecule B7-1. CONCLUSIONS: Although valve endothelial and interstitial cells express a similar range of cell-surface molecules, it is only the endothelial cells that are immunogenic. In addition, we have shown that these 2 cell types interact in a donor-specific manner to orchestrate the immune response and therefore may have clinical relevance in the allogeneic response of the heart valve recipients. PMID- 11436046 TI - Insulin-like growth factor 1 prevents neuronal cell death and paraplegia in the rabbit model of spinal cord ischemia. AB - OBJECTIVE: Insulin-like growth factor 1 has been shown to be cytoprotective against ischemia-reperfusion injury in various organs. However, spinal cord protection by insulin-like growth factor 1 has not been tested. We have therefore examined the effect of insulin-like growth factor 1 on neuronal cell death and motor function after spinal cord ischemia. METHODS: Japanese white rabbits were subjected to spinal cord ischemia by clamping the abdominal aorta for 15 minutes. Insulin-like growth factor 1 (0.3 mg/kg) at a dose equipotent to insulin (0.3 IU/kg) in lowering blood glucose level or the control (phosphate-buffered saline solution as a vehicle) was administered intravenously 30 minutes before the aortic clamp. RESULTS: Hind-limb motor function had recovered normally 48 hours after the operation in all the rabbits (n = 8) treated with insulin-like growth factor 1. In contrast, all the control-treated (n = 8) and all but one of the insulin-treated (n = 6) rabbits had deteriorated to paraplegia by 48 hours after the operation. Histopathologic sections in the involved spinal cord segment showed that a significantly (P <.0001) greater number of motor neuron cells were preserved in the rabbits treated with insulin-like growth factor 1 (17.9 +/- 4.8 per section) than in those treated with the control (8.0 +/- 2.1). Although insulin was equipotent to insulin-like growth factor 1 in preserving the number of motor neuron cells (18.5 +/- 2.7), the percentage of motor neuron cells positive for terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate-biotin nick-end labeling were significantly (P <.01) smaller in the rabbits treated with insulin-like growth factor 1 (6.0 +/- 4.6) compared with those treated with the control (54.6 +/- 33.8) and insulin (26.2 +/- 11.7). Immunohistochemical studies revealed that insulin-like growth factor 1 increased expression of the antiapoptotic Bcl-xL protein and inhibited expression of the proapoptotic Bax protein in motor neuron cells 24 and 48 hours after the operation. In contrast, expression of only Bax was increased after the operation in other groups of rabbits subjected to spinal cord ischemia. CONCLUSIONS: These results suggest that insulin-like growth factor 1, but not insulin with a conventional dose, protects motor neuron cells from ischemic spinal cord injury associated with differential regulation of Bcl-xL and Bax protein. PMID- 11436047 TI - Endovascular stent-grafting for traumatic aortic aneurysms with the use of a fenestrated stent-graft. PMID- 11436048 TI - The Ross procedure in children and young adults: a word of caution. AB - BACKGROUND: Disease of the aortic valve in children and young adults is a complex entity whose management is the subject of controversy. The Ross and the Ross Konno procedures have become the primary choices for aortic valve replacement in children because of growth potential, optimal hemodynamic performance, and lack of the need for anticoagulation. However, concern persists regarding the longevity of the pulmonary autograft, especially in patients with aortic insufficiency. METHODS: Between June 1993 and February 2000, 72 Ross and Ross Konno procedures were performed at our institution: 81% of the patients were less than 15 years old. Preoperative, postoperative, and follow-up clinical, echocardiographic, and hemodynamic data were reviewed. Statistical analysis was performed to identify the risk factors for deteriorating autograft function. RESULTS: Aortic insufficiency was an indication for the Ross procedure in 17 patients and mixed lesions with predominant aortic insufficiency in 10. Of the 45 other patients, 32 had aortic stenosis and 13 had mixed lesions with predominant aortic stenosis. There were no deaths during a follow-up of 5 to 80 months. Autograft reoperation was necessary in the follow-up period in 7 patients for severe aortic insufficiency. Moderate insufficiency was identified in 5 additional patients. Aortic insufficiency or predominant aortic insufficiency, as a preoperative hemodynamic indication for the Ross procedure, reached statistical significance (P =.031) as a risk factor for autograft failure. CONCLUSION: The Ross and the Ross-Konno procedures have changed the prognosis of children and young adults with complex aortic valve disease. However, the Ross procedure should be performed with caution in older children in whom aortic insufficiency is a preoperative hemodynamic indication. Further follow-up to delineate the risk factors for autograft dysfunction in children and young adults is necessary to better define the indications for the Ross procedure. PMID- 11436049 TI - Long-term results after early primary repair of tetralogy of Fallot. AB - OBJECTIVE: Early primary repair of tetralogy of Fallot has been routinely performed at Children's Hospital, Boston, since 1972. We evaluated the long-term outcome of this treatment strategy including the influence of a transannular patch. METHODS: Fifty-seven patients less than 24 months of age (median 8 months) underwent primary repair of tetralogy of Fallot between January 1972 and December 1977. Thirty-one patients had a transannular patch. Survival and freedom from reintervention were determined by the Kaplan-Meier method with 95% confidence intervals. RESULTS: There were 8 early deaths, and 1 patient died 24 years after initial repair. Recent follow-up was obtained for 45 of the 49 long-term survivors (92%). Median follow-up was 23.5 years. Ten patients underwent reintervention, 8 of whom underwent relief of right ventricular outflow tract obstruction. Right ventricular outflow tract obstruction occurred in 6 patients without a transannular patch and 2 with a transannular patch (33% vs 6%, P =.04). One pulmonary valve replacement was performed at another institution 20 years after the repair. Forty-one long-term survivors were in New York Heart Association class I and 4 were in class II. Actuarial survival was 86% at 20 years (95% confidence intervals = 80%-92%). Freedom from reintervention was 93% at 5 years (95% confidence intervals = 87%-99%) and 79% at 20 years (95% confidence intervals = 70%-86%). No significant differences were found between patients with and without a transannular patch (survival, P =.34; freedom from reintervention, P =.09, log-rank tests). CONCLUSIONS: Long-term survival is excellent and the freedom from reintervention is satisfactory after early primary repair of tetralogy of Fallot in the 1970s. Use of a transannular patch does not reduce late survival and is associated with a lower incidence of right ventricular outflow tract obstruction. PMID- 11436050 TI - Valve-sparing operation for balloon-induced aortic regurgitation in congenital aortic stenosis. AB - OBJECTIVE: Aortic regurgitation after balloon dilation of congenital aortic stenosis may be treated with valve repair as an alternative to replacement. METHODS: Charts and echocardiograms of all patients undergoing aortic valve operations after balloon dilation of congenital aortic stenosis at our institution between January 1988 and December 1999 were reviewed. RESULTS: Twenty one patients underwent valvuloplasty for predominant aortic regurgitation 9 months to 15 years (mean, 6.1 years) after balloon dilation. The mean +/- SD age at the time of the operation was 11 +/- 7 years. Aortic regurgitation was caused by a combination of commissural avulsion (10), cusp dehiscence with retraction (9), cusp tear (5), central incompetence (2), perforated cusp (1), or cusp adhesion to the aortic wall (1). Repair techniques included commissural reconstruction with a pericardial patch (8), pericardial patch cusp augmentation (6), primary suture repair (6), raphae release and debridement (4), commissurotomy (4), commissural resuspension with sutures (3), and cusp release (1). There were no deaths. At a mean follow-up of 30.1 months (range, 9 months-8 years), all patients were asymptomatic, and the grade of aortic regurgitation had been significantly reduced (P <.001). Left ventricular end-diastolic dimension z scores and proximal regurgitant jet/aortic anulus diameter ratios were significantly reduced (P <.001) and remained so over time. Freedom from reoperation for late failure was 100%, and overall freedom from reintervention was 80% at 3 years. CONCLUSION: Aortic valve repair for balloon-induced aortic regurgitation is reproducible and durable at medium-term follow-up. PMID- 11436051 TI - Pulmonary atresia with ventricular septal defect and persistent airway hyperresponsiveness. AB - OBJECTIVE: We and others have observed significant hyperinflation and airflow obstruction after the surgical repair of pulmonary atresia and ventricular septal defect. This study sought to objectively characterize this problem and determine the prevalence of airway hyperresponsiveness in these patients. METHODS: We performed a prospective study of children and young adults with pulmonary atresia and ventricular septal defect between June 1996 and December 1998. The participants were stratified into 2 distinct molecular genotypes on the basis of chromosome 22q11.2 microdeletion. A clinical diagnosis of asthma and an objective assessment of airway hyperresponsiveness were determined by means of an asthma inventory scale and methacholine challenge testing, respectively. Thirty-three patients were enrolled. Thirteen had velocardiofacial syndrome, each with chromosome 22q11.2 microdeletion. RESULTS: None of the nonsyndromic patients had evidence for haploinsufficiency. Overall, 66.7% (22/33) met criteria for a clinical diagnosis of airway hyperresponsiveness: 62% (8/13) from the microdeletion genotype and 70% (14/20) from the nonsyndromic group. CONCLUSIONS: We have identified an extremely strong association between pulmonary atresia and ventricular septal defect and persistent airway hyperresponsiveness. Haploinsufficiency at chromosome 22q11.2 did not contribute to this predilection for airway hyperresponsiveness. These results provide a basis to anticipate persistent respiratory difficulties after operations in patients with pulmonary atresia and ventricular septal defect. Moreover, this at-risk patient population may yield unique insights into fundamental mechanisms involved in the pathogenesis of airway hyperresponsiveness. PMID- 11436052 TI - Anatomic assessment of hand circulation in harvesting the radial artery. PMID- 11436053 TI - Prenatal right ventricular infarction associated with hypoplastic left heart syndrome. PMID- 11436054 TI - Angioplasty balloon occlusion of internal thoracic artery in redo surgery in patients with coronary artery bypass operations. PMID- 11436055 TI - Tracheal laceration: a rare complication of median sternotomy. PMID- 11436056 TI - Endovascular stent-graft repair for thoracic aortic aneurysm associated with right-sided aortic arch. PMID- 11436057 TI - Early failure of the autograft valve after the Ross procedure. PMID- 11436058 TI - Staged perfusion with an axillary artery graft and deep hypothermia during descending aortic replacement. PMID- 11436059 TI - Clinical utility of the platelet function analyzer (PFA-100) in cardiothoracic procedures involving extracorporeal circulation. PMID- 11436060 TI - Mixed venous oxygen saturation does not adequately predict cerebral perfusion during pediatric cardiopulmonary bypass. PMID- 11436061 TI - Elevated plasma concentration of vascular endothelial growth factor in cardiac myxoma. PMID- 11436062 TI - Replacement of infected prosthesis on the ascending aorta with an abdominal aortic autograft in a young patient. PMID- 11436063 TI - Successful experience with simultaneous lung volume reduction and cardiac procedures. PMID- 11436064 TI - A novel approach to tracheal replacement: the use of an aortic graft. PMID- 11436065 TI - Sudden intestinal necrosis one month after acute aortic dissection. PMID- 11436066 TI - Presidential address: vascular surgery and the Midwestern Vascular Surgical Society in the new millennium. PMID- 11436067 TI - Reoperation for recurrent carotid stenosis: early results and late outcome in 199 patients. AB - PURPOSE: This study was undertaken to determine the safety and efficacy of reoperations for recurrent carotid stenosis (REDOCEA) at the Cleveland Clinic. MATERIALS AND METHODS: From 1989 to 1999, 206 consecutive REDOCEAs were performed in 199 patients (131 men, 68 women) with a mean age of 68 years (median, 69 years; range, 47-86 years). A total of 119 procedures (57%) were performed for severe asymptomatic stenosis, 55 (27%) for hemispheric transient ischemic attacks or amaurosis fugax, 26 (13%) for prior stroke, and 6 (3%) for vertebrobasilar symptoms. Eleven REDOCEAs (5%) were combined with myocardial revascularization, and another 19 (9%) represented multiple carotid reoperations (17 second reoperations and 2 third reoperations). Three REDOCEAs (1%) were closed primarily, and nine (4%) required interposition grafts, whereas the remaining 194 (95%) were repaired with either vein patch angioplasty (139 [68%]) or synthetic patches (55 [27%]). Three patients (2%) were lost to follow-up, but late information was available for 196 patients (203 operations) at a mean interval of 4.3 years (median, 3.9 years; maximum, 10.2 years). RESULTS: Considering all 206 procedures, there were 7 early (< 30 days) postoperative neurologic events (3.4%), including 6 perioperative strokes (2.9%) and 1 occipital hemorrhage (0.5%) on the 12th postoperative day. Seventeen additional neurologic events occurred during the late follow-up period, consisting of eight strokes (3.9%) and nine transient ischemic attacks (4.4 %). With the Kaplan-Meier method, the estimated 5-year freedom from stroke was 92% (95% CI, 88%-96%). There were two early postoperative deaths (1%), both from cardiac complications after REDOCEAs combined with myocardial revascularization procedures. With the Kaplan-Meier method, the estimated 5-year survival was 81% (range, 75%-88%). A univariate Cox regression model yielded the presence of coronary artery disease as the only variable that was significantly associated with survival (P =.024). The presence of pulmonary disease (P =.036), diabetes (P =.01), and advancing age (P =.006) was found to be significantly associated with stroke after REDOCEA. Causes of 53 late deaths were cardiovascular problems in 25 patients (47%), unknown in 14 (26%), renal failure in 4 (8%), stroke in 3 (6%), and miscellaneous in 7 (13%). CONCLUSIONS: We conclude that REDOCEA may be safely performed in selected patients with recurrent carotid stenosis and that most of these patients enjoy long-term freedom from stroke. PMID- 11436068 TI - Timing of postcarotid complications: a guide to safe discharge planning. AB - OBJECTIVES: Currently, our standard of practice is that patients undergoing carotid endarterectomy (CEA) may be safely discharged on the first postoperative day. Because many patients do not appear to require overnight observation, we wanted to determine the safety and feasibility of same-evening discharge by establishing the timing of postoperative complications, which may potentially require operative intervention. METHODS: A total of 835 consecutive patients undergoing CEA were retrospectively reviewed. Sixty-two patients had a postoperative wound hematoma or neurologic deficit (ND) (transient ischemic attack or stroke) within 24 hours of their operation, complications potentially requiring a second operation. Excluded were 64 patients not eligible for same-day discharge because of other reasons (eg, heparinization, CEA with coronary artery bypass grafting). RESULTS: Sixty-two patients (8.0%) had ND (26 [3.4%]) or neck hematoma (NH) (36 [4.7%]) within 24 hours of their CEA. Nineteen (73%) of the NDs were diagnosed in the operating room or recovery room, 5 (19%) within 8 hours of the operation, and 2 (7.7%) after 8 hours but in less than 24 hours. Of the NHs, 23 (66%) were diagnosed in the recovery room, 11 (31%) within 8 hours, and 1 (2.7%) after 8 hours. Of the outliers, one patient experienced a blowout of the vein graft occurring on postoperative day 1, one patient had a delayed ipsilateral stroke, and one had a vertebrobasilar stroke. Overall, only three of 773 (0.4%) patients undergoing CEA had a complication occurring more than 8 hours after operation. CONCLUSION: NDs and NHs in post-CEA patients occurred within 8 hours of operation in 95% of those patients experiencing these complications or 99.6% of all CEA patients. These data indicate that same-evening discharge may be safely performed without increasing the adverse effects of stroke or hematoma. This plan has cautiously been initiated at this institution. PMID- 11436069 TI - Lepirudin is a safe and effective anticoagulant for patients with heparin associated antiplatelet antibodies. AB - OBJECTIVE: The purpose of this study was to determine whether lepirudin, a direct thrombin inhibitor, is a safe and effective anticoagulant for patients with heparin-associated antiplatelet antibodies (HAAbs). METHODS: The charts of HAAb positive patients who received lepirudin were reviewed. Lepirudin use was analyzed for indication, duration, and effectiveness of anticoagulation, and for adverse events. HAAb presence was determined by platelet aggregation. RESULTS: Eighteen HAAb-positive patients received lepirudin: 9 had previous documentation of HAAb, 6 had thrombocytopenia while receiving heparin; and 3 had HAAb after a thrombotic event. The indications for lepirudin anticoagulation included thromboembolism prophylaxis (5), arterial thromboses (5), pulmonary embolus (3) or deep venous thrombosis (1), and one each for atrial fibrillation, myocardial infarction, artificial heart valves, and hemodialysis access. The average duration of therapy was 4.04 days. Fifteen patients achieved adequate anticoagulation (activated partial thromboplastin time [aPTT] ratio > 2.0) with lepirudin. Seven patients had aPTTs that were sometimes supratherapeutic (aPTT > 100 seconds) but did not bleed. In all patients who had heparin-induced thrombocytopenia, platelet counts were normalized while they received lepirudin. There were two complications: one patient fell and had a calf hematoma (aPTT ratio 3.24), and one patient who received lepirudin during nine separate hospitalizations had epistaxis (aPTT ratio 2.86) during her ninth hospitalization. Another patient received lepirudin during two hospitalizations without an adverse event. CONCLUSION: Lepirudin is a safe and effective anticoagulant for patients with HAAbs. The platelet counts of all patients with heparin-induced thrombocytopenia were normalized while they received lepirudin. Careful monitoring of the aPTT and avoidance of trauma while patients are receiving lepirudin are recommended. PMID- 11436070 TI - Bedside vena cava filter placement guided with intravascular ultrasound. AB - OBJECTIVE: The purpose of this study was to report a feasibility trial approved by the Institutional Review Board for insertion of inferior vena cava (IVC) filters with intravascular ultrasound (IVUS) guidance in the intensive care unit. METHODS: Between October 1998 and May 2000, 26 patients (15 men, 11 women; age range, 22-86 years; mean, 55 years) were enrolled. Eight patients (31%) underwent prophylactic filter placement, and 18 patients (69%) had venous thromboembolism (deep venous thrombosis = 16, pulmonary embolism = 2) with contraindications to anticoagulation. A single groin puncture was used for IVUS and filter placement. Location of major branch veins, thrombosis, and caval diameter were readily demonstrated without the use of radiocontrast agents. Mapping of the IVC permitted assessment of ideal filter location. Postprocedure radiographs (23 of 26) were obtained to document filter position. Seventeen of 26 had follow-up lower extremity duplex studies. RESULTS: Twenty-four (92%) of 26 patients underwent successful filter deployment. The two other patients had filters subsequently placed by means of traditional fluoroscopic techniques. One femoral vein insertion site thrombosis resolved after a month. One patient experienced symptomatic caval thrombosis thought to be caused by thrombus trapping 55 days after the procedure. No pulmonary emboli occurred after filter placement. One patient's death was unrelated to vena cava filter placement. The hospital charge for bedside filters was $3623 compared with $4165 (P =.281) for fluoroscopic placement. CONCLUSION: Bedside insertion of an IVC filter with IVUS guidance is feasible and may be an effective alternative in the intensive care unit. No additional costs were incurred in this small series. Protocol refinements should reduce the incidence of complications. The results of this study support the need for further evaluation comparing it with standard techniques. PMID- 11436071 TI - Long-term results justify autogenous infrainguinal bypass grafting in patients with end-stage renal failure. AB - INTRODUCTION: Infrainguinal bypass grafting for limb-threatening ischemia in patients with end-stage renal disease is generally thought to be associated with increased operative risk and poor long-term outcome. This retrospective study was undertaken to examine the modern-era, long-term results of infrainguinal bypass grafting in dialysis-dependent patients. METHODS: Over the past 5 years in a single institution, 425 lower extremities (368 consecutive patients) were revascularized for the indication of limb salvage. Sixty-four patients (82 limbs) were dialysis-dependent at the time of revascularization, and this group was analyzed separately. They exhibited statistically significant higher incidences of diabetes (83% vs 56%; P <.001), hypertension (91% vs 74%; P <.001), and more distal vascular disease, which required a greater proportion of proximal anastomoses at the popliteal level (24% vs 11%; P <.01) and distal anastomoses at the infrapopliteal level (75% vs 65%; P <.05). RESULTS: Despite the higher prevalence of comorbid conditions and distal disease in patients with renal failure, their perioperative 30-day mortality rate remained low (4.9%) and was not significantly different from that in patients with functioning kidneys (2.9%; P = not significant). After a median follow-up of 11 months (range, 0-60 months), the 3-year autogenous conduit secondary graft patency in patients with renal failure was no different than in patients with functioning kidneys (67% +/- 9% vs 64% +/- 5%; P = not significant). Nonautogenous conduits in dialysis-dependent patients exhibited a significantly poorer outcome with only 27% +/- 12% remaining secondarily patent at 2 years. As expected, both limb salvage and patient survival were significantly less in patients with renal faiture, although both exceeded 50% at 3 years (limb salvage 59% +/- 8% vs 68% +/- 5%; P <.05; patient survival 60% +/- 8% vs 86% +/- 4%; P <.001). The often-quoted phenomenon of limb loss, despite a patent bypass graft, occurred infrequently in this study (n = 3 of 82 limbs). CONCLUSION: Infrainguinal revascularization can be performed in dialysis-dependent patients with acceptable perioperative and long-term results, especially in patients in whom adequate autologous conduit is available. PMID- 11436072 TI - Efficacy and durability of autogenous saphenous vein conduits for lower extremity arterial reconstructions in preadolescent children. AB - OBJECTIVE: Limb length discrepancies (LLDs) in growing children may accompany extremity arterial occlusions. Revascularization with vein grafts has been questioned because of degenerative graft changes observed at other sites. This study was undertaken to define vein graft durability and efficacy in lower extremity revascularizations in preadolescent children. STUDY DESIGN: Fourteen children (10 boys, 4 girls) with a mean age of 7.3 years (range, 2-11 years) who underwent 16 lower extremity revascularizations with greater saphenous vein grafts were subjected to follow-up with graft ultrasonography, ankle/brachial indices (ABIs) with and without exercise, and limb length determinations. A mean of 5.7 years elapsed between the onset of ischemia and operation. Arterial occlusions resulted from cardiac catheterizations (11), arteritis (1), dialysis cannulation (1), and penetrating trauma (1). Indications for operation included LLD (6), claudication (4), both LLD and claudication (3), markedly diminished ABIs with a potential for LLD (2), and a traumatic transection with hemorrhage (1). The reconstructions with 15 reversed and one in situ vein grafts included iliofemoral (11), femorofemoral (1), aortofemoral (1), femoropopliteal (1), popliteal-popliteal (1), and popliteal-posterior tibial (1) arterial bypass grafts. RESULTS: Among patent grafts available for follow-up, 36% (5 of 14) remained unchanged, 50% (7 of 14) developed nonaneurysmal dilatation, and 14% (2 of 14) exhibited nonprogressive aneurysmal expansion. One graft became occluded, and one graft was lost to follow-up. Collectively, the grafts manifest an 11.2% expansion at an average of 10.7 years postoperatively. ABIs increased from 0.75 preoperatively to 0.97, at an average of 11.0 years postoperatively. LLDs were reduced from 1.66 to 1.24 cm, at an average of 11.4 years postoperatively. CONCLUSION: Vein graft reconstructions of lower extremity arteries in preadolescent children are durable. They provide an efficacious means of restoring normal blood flow, and in 70% of children their preexisting LLDs were reduced. PMID- 11436073 TI - Ruptured abdominal aortic aneurysms: the excessive mortality rate of conventional repair. AB - OBJECTIVE: Rupture of abdominal aortic aneurysms (AAAs) remains lethal. In a report of patients treated in the 1980s, we recommended aggressive management. Our continued experience prompted us to reevaluate this policy. METHODS: We reviewed clinical variables affecting outcome, morbidity, mortality, and trends in mortality of all patients managed at our institution with ruptured AAAs between January 2, 1980, and November 30, 1998. RESULTS: The study group included 413 consecutive patients, 339 men and 74 women. The mean age was 74.3 years (range, 49-96); 116 (28%) patients were older than 80 years. AAA was diagnosed before rupture in 119 (29%) patients. Eighty (19%) patients had preoperative cardiac arrest. Twenty-nine (7%) patients died before operation; 65 (17%) died during the operation. The surgical mortality rate (30-day) was 37%; the overall mortality rate was 45% and was higher in women (68%) than in men (40%) (P <.001). Advanced age, APACHE (Acute Physiology and Chronic Health Evaluation) II score, initial hematocrit, and preoperative cardiac arrest were associated multivariately with 30-day mortality rates by means of stepwise logistic regression (P <.05). Twelve (23%) of 53 patients with cardiac arrest survived the operation. Logistic regression, adjusted for age, sex, and APACHE II score, demonstrated a decrease in overall and 30-day mortality rates (P <.001) over 18 years. The mean overall mortality rate was 51% from 1980 to 1984 and 42% from 1994 to 1998. CONCLUSIONS: The mortality rate of ruptured AAAs remains excessive, despite improvement over 18 years. Patients older than 80 years with shock or cardiac arrest have the highest mortality rate and should be evaluated for possible endovascular treatment. Because the diagnosis of AAA was unknown in more than 70% of patients, screening of the high-risk population and elective repair are recommended. PMID- 11436074 TI - Minimal incision aortic surgery. AB - PURPOSE: In this study we evaluated the clinical and economic impact of minimal incision aortic surgery (MIAS) for treatment of patients with abdominal aortic aneurysms (AAAs) and aortoiliac occlusive disease (AIOD). METHOD: Fifty patients with either AAA (34) or AIOD (16), prospectively treated with the MIAS technique, were compared with 50 patients (40 AAA and 10 AIOD) treated in the same time period with long midline incision and extracavitary small bowel retraction. MIAS was also compared with a cohort of 32 patients with AAA treated by means of endoaortic stent-grafts. Outcomes and cost (based on metric mean length of stay) were compared for the open and endoaortic techniques. RESULTS: Patients who experienced no perioperative complications after the MIAS or endovascular repair technique had shorter hospital stays than patients with uncomplicated aortic repairs performed with a traditional long midline abdominal incision (3 days vs 3 days vs. 7.2 days). Hospital stay was also significantly shorter for the less invasive procedures when perioperative complications were included (4.8 days vs. 4.3 days vs 9.3 days). The MIAS and endovascular aortic repair groups had a shorter intensive care unit stay (< or = 1.0 day) and a quicker return to general dietary feeding (2.5 days) than patients treated with standard open repair (1.8 days, 4.7 days). The overall morbidity for the MIAS technique (14%) and endovascular technique (21%) was not significantly different from standard open repair (24%). The mortality rate for the different treatment groups was equivalent (MIAS, 2%; endovascular repair, 3%; standard repair, 2%). The MIAS was more cost-efficient than standard open repair ($12,585 vs $18,445) because of shorter intensive care unit and hospital stay and was more cost-efficient than endoaortic repair ($12,585 vs $32,040) because of reduced, direct intraoperative costs. CONCLUSIONS: MIAS is as safe as standard open or endovascular repair in the treatment of AAA and AIOD. MIAS is more cost-efficient than standard open or endoaortic repair. PMID- 11436075 TI - The outcome in the United States after thoracoabdominal aortic aneurysm repair, renal artery bypass, and mesenteric revascularization. AB - OBJECTIVES: The purpose of this study was to determine outcome and identify predictors of death after thoracoabdominal aortic aneurysm (TAA) repair, renal artery bypass (RAB), and revascularization for chronic mesenteric ischemia (CMI). PATIENTS AND METHODS: In this retrospective analysis, data were obtained from the Nationwide Inpatient Sample, a 20% all-payer stratified sample of hospitals in the United States during 1993 to 1997. Patients were identified by the presence of a diagnostic or procedure code from the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM). The main outcomes we examined were death, ICD-9-CM -based complications, length of stay, hospital charges, and disposition. A multivariate model was constructed to predict death. RESULTS: A total of 2934 patients were identified (TAA, 540; RAB, 2058; CMI, 336) in the database. The mean age was comparable (TAA, 69 +/- 9 years; RAB, 66 +/- 12 years; CMI, 66 +/- 11 years), but the breakdown between the sexes varied by procedure (male: TAA, 53%; RAB, 55%; CMI, 24%). The mortality rate (TAA, 20.3%; RAB, 7.1%; CMI, 14.7%), complication rate (TAA, 62.2%; RAB, 37.4%; CMI, 44.6%), and the percentage of patients discharged to another institution (TAA, 21.2%; RAB, 9.3%; CMI, 12.0%) were clinically significant for all procedures. The mortality rate for RAB was greater when performed concomitant with an aortic reconstruction (4.4% vs 8.3%). All three procedures were resource intensive as reflected by the median length of stay (TAA, 14 days; RAB, 9 days; CMI, 14 days) and median hospital charges (TAA, $64,493; RAB, $36,830; CMI, $47,390). The multivariate model identified several variables for each procedure that had an impact on the predicted mortality rate (TAA, 14%-76%; RAB, < 1%-46%; CMI, < 2% 87%). CONCLUSIONS: The operative mortality rates across the United States for patients undergoing TAA repair and RAB are greater than commonly reported in the literature and mandate reexamining the treatment strategies for these complex vascular problems. PMID- 11436076 TI - A prospective study of discharge disposition after vascular surgery. AB - OBJECTIVE: The purpose of this study was to determine what factors are predictive of a decline in independent living after vascular surgery during recovery. METHODS: Demographics, risk factors, operations, complications, wound status, and discharge disposition for all patients admitted to a tertiary vascular surgery service for any surgical procedure were prospectively recorded at the time of discharge. The declining order of dispositions at discharge were home (no professional assistance), home (professional assistance), rehabilitation facility, and skilled nursing facility. RESULTS: Over a 15-month period, 380 patients underwent 442 primary operations. Primary operations included 74 (17%) carotid procedures, 38 (8%) aortic procedures, 186 (42%) extremity revascularizations, 29 (7%) major amputations, 45 (10%) minor amputations, and 70 (16%) other. There were 148 (33%) complications and 85 (20%) subsequent operations (same hospitalization); 159 (36%) open wounds occurred. Forty-six percent of the patients were discharged to home (no professional assistance), 28% to home (professional assistance), 3% to a rehabilitation facility, and 18% to a skilled nursing facility; 5% died. At discharge, 51% of patients required professional assistance, 39% had a decline in disposition, and 12% went from home (+/- professional assistance) to a facility. By multivariate regression analysis, a hospital stay more than 6 days, emergency operation, open operative wound, systemic complications, and minor amputation were significantly associated (P <.001) with a decline in disposition at discharge (odds ratios: 5.5, 3.7, 3.6, 3.6, and 2.8, respectively). CONCLUSIONS: Prospective study reveals that a large proportion of patients (39%) had a decline in disposition after vascular surgery. A hospital stay more than 6 days, emergency operation, open operative wound, systemic complications, and minor amputation were strong independent predictors of decline. This information suggests modifications in treatment strategies may improve independent living status after vascular surgery and decrease the intense use of extended care resources required for this patient population during recovery. PMID- 11436077 TI - Ancillary techniques to facilitate endovascular repair of aortic aneurysms. AB - PURPOSE: The ability to treat abdominal aortoiliac aneurysms and thoracic aortic aneurysms may be limited by coexisting arterial disease. Device deployment may be impaired by occlusive disease and tortuosity of the arteries used to access the aneurysm or by suitability of the implantation sites. In this study we describe the auxiliary procedures performed to circumvent these obstacles and thereby enable endovascular aneurysm repair. PATIENTS AND METHODS: Between January 1, 1993, and December 31, 1999, 390 patients treated for aneurysm of the aorta with endovascular devices were entered prospectively in a vascular registry. Fifty (12%) of the 390 patients required adjunctive surgical techniques to (1) create or extend the length of the proximal or distal device implantation site or (2) permit device navigation through diseased iliac arteries. Auxiliary techniques used to extend or enhance implantation sites were elephant trunk graft (n = 2), the construction of renovisceral bypass grafts (n = 1), and subclavian artery transposition (n = 2). Plication of the common iliac artery at its bifurcation was performed in conjunction with femorofemoral bypass graft in nine patients to allow preservation of pelvic circulation by avoiding internal iliac artery sacrifice. Construction of a bypass graft to transpose the internal iliac artery orifice was performed in one patient. The auxiliary techniques used to facilitate device navigation were iliac artery angioplasty or stenting (n = 8), external iliac artery endovascular endarterectomy or straightening (n = 14), endoluminal iliofemoral bypass conduit (n = 5), and the construction of an open iliofemoral bypass conduit (n = 8). RESULTS: Successful deployment of the endovascular devices was achieved in 49 (98%) of 50 patients. Auxiliary techniques were successful in providing access for endovascular device deployment in all 35 patients (100%). Mean follow-up for techniques to facilitate device navigation is 26 months for endovascular procedures and 42 months for the open bypass graft construction patients; no occlusions were observed at this moment. There were five patients with incisional hematomas that did not necessitate intervention. Fourteen (94%) of 15 patients underwent successful device implantation after the auxiliary maneuvers to enhance implantation site. Mean follow-up for implantation site manipulation is 28 months. One of the subclavian transpositions had a new onset of Horner's syndrome, two of nine patients who had common iliac artery ligated had retroperitoneal hematomas that did not necessitate interventions, and no colon ischemia was seen. The patient who underwent nonanatomic bypass grafting of viscero-renal arteries had a retroperitoneal hematoma that necessitated reexploration. CONCLUSIONS: Significant coexisting arterial disease may be encountered in patients with aortic or iliac aneurysms. Identification of coexisting arterial diseases is essential to help tailor the appropriate supplemental surgical procedure to allow the performance of endovascular aneurysm repair in patients who would otherwise require open surgical repair. PMID- 11436078 TI - Nitric oxide inhibition increases matrix metalloproteinase-9 expression by rat aortic smooth muscle cells in vitro. AB - OBJECTIVE: The hypothesis to be tested was that diminished bioavailable nitric oxide (NO) affects matrix metalloproteinase (MMP) expression and activation in vascular smooth muscle cells (SMCs). METHODS: Cultivated rat aortic SMCs (RA SMCs) were exposed to increasing concentrations of L-N-monomethyl arginine (L NMMA), a nonselective inhibitor of NO synthase, in the presence of proinflammatory cytokines (50 ng/mL interleukin [IL]-1beta, 50 ng/mL interferon gamma, and 30 microg/mL lipopolysaccharide). Nitrite and nitrate, two of the final end products of NO metabolism, were measured in media collected at 48 hours with the use of the Saville assay (n = 4). MMP activity was measured with 1% gelatin zymography (n = 4). In separate experiments in which 2 ng/mL of IL-1beta and L-NMMA was used, MMP protein and messenger RNA (mRNA) levels were determined with Western blot analysis (n = 3) and semiquantitative reverse transcriptase polymerase chain reaction (n = 3), respectively. Data were analyzed with nonparametric analysis of variance. RESULTS: Increasing concentrations of the NO synthase inhibitor L-NMMA caused a dose-dependent decrease (P <.05) in nitrite and nitrate production by RA-SMCs after cytokine exposure. Zymography documented an early dosedependent increase (P <.05 compared with cytokines alone) in 92-kd MMP activity, with no significant changes in 72-kd MMP activity after treatment with L-NMMA (P >.05 compared with cytokines alone). Reverse transcriptase polymerase chain reaction and Western blot analysis revealed that the addition of L-NMMA to IL-1beta-stimulated RA-SMCs led to significant increases in MMP-9 mRNA (n = 3, P <.01 for 1.0 mmol/L L-NMMA) and MMP-9 protein levels (n = 3, P <.05), respectively. No differences in MMP-2 mRNA or protein levels were demonstrated. CONCLUSIONS: Inhibition of cytokine-induced NO expression in RA-SMCs is associated with a selective, dose-dependent increase in MMP-9 expression and synthesis. These findings suggest that alterations in local NO synthesis may influence MMP-9-dependent vessel wall damage. PMID- 11436079 TI - Genetic similarity in inflammatory and degenerative abdominal aortic aneurysms: a study of human leukocyte antigen class II disease risk genes. AB - PURPOSE: Clinically, abdominal aortic aneurysms (AAAs) display a spectrum of inflammation that extends from apparently noninflamed (degenerative) AAAs to the classic inflammatory variant. Genes encoded in the human leukocyte antigen (HLA) region are important in the development of both variants of AAA; however, their role in progression to the inflammatory variant is unknown. The purpose of this study was to compare HLA class II genes in patients with degenerative versus classic inflammatory AAAs and to quantify their impact as disease risk factors. METHODS: Genotypes of the 12 major alleles of the HLA-DR B1 locus were determined in patients with degenerative (102) and inflammatory (40) AAAs who were compared with controls (118). Univariate and multivariate logistic regression analyses were used to determine allele distributions and to quantify disease risk. RESULTS: Distribution of the HLA-DR B1 alleles was nonrandom and similar in both degenerative and inflammatory AAA groups compared with controls. The B1*02 and B1*04 alleles were enhanced in both degenerative (39.2% vs. 25.4%, P =.03; and 35.3% vs. 24.6%, P =.08 respectively) and inflammatory (47.5% vs. 25.4%, P =.01; and 32.5% vs. 24.6%, P =.09, respectively) AAAs compared with controls. The B1*02 and B1*04 alleles were associated with risk for both degenerative (odds ratio [OR] 2.2; 95% CI, 1.2-4.0; and OR 2.0; 95% CI, 1.1-3.7, respectively) and inflammatory AAAs (OR 3.7; 95% CI, 1.8-8.6; and OR 2.5; 95% CI, 1.1-6.1). CONCLUSION: This study demonstrates that identical HLA alleles function as genetic risk factors for both inflammatory and degenerative AAAs. These results support the concept of a common, immune-mediated pathogenesis for AAAs that may be modulated by HLA-independent factors. PMID- 11436080 TI - The effects of extremely low shear stress on cellular proliferation and neointimal thickening in the failing bypass graft. AB - OBJECTIVE: Previous studies demonstrating a correlation between low shear stress (tau = 5-15 dyne/cm(2)) and experimental vein graft neointimal thickening (NIT) support the role of low tau in vein graft failure. However, a simple linear relationship between low tau and NIT would underestimate the degree of NIT evident in high-grade occlusive lesions of failing human vein grafts. In this study we used a new experimental model that maintains patency at low tau (< 2 dyne/cm(2)), to delineate possible deviations from linearity in the low tau --> NIT hypothesis. METHODS: Thirty-two New Zealand White rabbits underwent creation of a common carotid vein patch with a segment of ipsilateral external jugular vein. Very low tau was created in 13 patches by ligation of the distal common carotid artery, leaving the only outflow through a small muscular branch. Normal tau was created in 11 patches by leaving the common carotid artery outflow intact. High tau was created in eight patches by ligation of the contralateral common carotid artery. Six patches were harvested after 2 weeks for measurement of cell cycle entry by proliferating cell nuclear antigen (PCNA) immunohistochemistry. The remaining 26 patches were harvested after 4 weeks, perfusion fixed, and excised for morphometric analysis. RESULTS: Mean blood flow and tau at implantation ranged from 0.5 to 41 mL/min and 0.07 to 15 dyne/cm(2), respectively. At the time of harvest, 30 of 32 patches remained patent, and the artificially created aberrations in blood flow were maintained (range, 0.7-41 mL/min). After 2 weeks PCNA immunohistochemistry showed a significantly higher level of cell cycling in patches exposed to low tau (40 +/- 5 vs 1.6 +/- 0.3 PCNA positive cells per high-power field; P <.001), which is equivalent to approximately 20% of the total cells present. In patches harvested after 4 weeks, NIT ranged from 42 to 328 microm and significantly correlated with mean tau at implantation. Patches with very low tau exhibited histologic characteristics similar to those of failing human bypass grafts, including laminar thrombus and flow-limiting luminal stenosis. The relationship between tau and NIT was nonlinear in that extremely low tau (< 2 dyne/cm(2)) resulted in NIT beyond that predicted by a simple linear correlation (P =.003). CONCLUSION: Extremely low tau (< 2 dyne/cm(2)) stimulates high rates of smooth muscle cellular proliferation in arterialized vein patches. NIT is accelerated in these regions of low tau far beyond that predicted by a simple linear model. The nonlinear nature of the cellular proliferative response and NIT at tau less than 2 dyne/cm(2) may explain the rapid progression of neointimal lesions in failing bypass grafts. PMID- 11436081 TI - Endoleak after endovascular repair of abdominal aortic aneurysm. AB - PURPOSE: We sought to assess the role of endovascular techniques in the management of perigraft flow (endoleak) after endovascular repair of an abdominal aortic aneurysm. METHOD: We performed endovascular repair of abdominal aortic aneurysm in 114 patients, using a variety of Gianturco Z-stent-based prostheses. Results were evaluated with contrast-enhanced computed tomography (CT) at 3 days, 3 months, 6 months, 12 months, and every year after the operation. An endoleak that occurred 3 days after operation led to repeat CT scanning at 2 weeks, followed by angiography and attempted endovascular treatment. RESULTS: Endoleak was seen on the first postoperative CT scan in 21 (18%) patients and was still present at 2 weeks in 14 (12%). On the basis of angiographic localization of the inflow, the endoleak was pure type I in 3 cases, pure type II in 9, and mixed pattern in 2. Of the 5 type I endoleaks, 3 were proximal and 2 were distal. All five resolved after endovascular implantation of additional stent-grafts, stents, and embolization coils. Although inferior mesenteric artery embolization was successful in 6 of 7 cases and lumbar embolization was successful in 4 of 7, only 1 of 11 primary type II endoleaks was shown to be resolved on CT scanning. There were no type III or type IV endoleaks (through the stent-graft). Endoleak was associated with aneurysm dilation two cases. In both cases, the aneurysm diameter stabilized after coil embolization of the inferior mesenteric artery. There were two secondary (delayed) endoleaks; one type I and one type II. The secondary type I endoleak and the associated aneurysm rupture were treated by use of an additional stent-graft. The secondary type II endoleak was not treated. CONCLUSIONS: Type I endoleaks represent a persistent risk of aneurysm rupture and should be treated promptly by endovascular means. Type II leaks are less dangerous and more difficult to treat, but coil embolization of feeding arteries may be warranted when leakage is associated with aneurysm enlargement. PMID- 11436082 TI - Diagnosis and endovascular treatment of iliocaval compression syndrome. AB - PURPOSE: The purpose of this study was to evaluate the clinical presentation, diagnosis, and endovascular treatment of iliocaval compression syndrome (ICS). PATIENTS AND METHODS: During a 3-year period, 18 patients (17 women, 1 man; mean age, 42 years) presented with clinical and imaging findings consistent with ICS. All patients were evaluated with venography and Doppler ultrasound (DUS), 13 of 18 with intravascular pressure measurements, 12 of 18 with intravascular ultrasound, 9 of 18 with air plethysmography (APG), and 4 of 18 with magnetic resonance venography. Seventeen patients were treated with endovascular stenting, one was treated with angioplasty alone, and six received adjunct thrombolysis. RESULTS: Despite the presence of stenosis or occlusion in all cases, APG indicated no iliac vein obstruction (outflow fraction > or = 40%) in nine patients. DUS revealed acute (6) or chronic (7) unilateral iliofemoral deep venous thrombosis in 13 of 18 patients, whereas the results of five of 18 DUS studies were normal. Recanalization and stent placement (n = 17) or angioplasty (n = 1) was achieved in all patients. The average pressure gradient was 5.6 mm Hg preprocedure and 0.6 mm Hg postprocedure. The primary patency rate demonstrated with DUS (n = 17) and venography (n = 7) at 6 months was 89%. The primary patency rate at 12 months was 79%. CONCLUSIONS: ICS often presents as sudden unilateral left lower extremity pain and swelling in young to middle-aged female patients after pregnancy, surgery, or a period of inactivity. Venography, intravascular ultrasound, and magnetic resonance venography demonstrate high sensitivity, whereas APG-outflow fraction demonstrates low sensitivity in the diagnosis of ICS. Endovascular stenting and angioplasty provide safe and effective early and intermediate-term treatment of symptomatic ICS. PMID- 11436083 TI - Percutaneous transluminal angioplasty of the femoropopliteal arteries in limbs with chronic critical lower limb ischemia. AB - PURPOSE: The aim of the study was to evaluate the results of percutaneous transluminal angioplasty (PTA) of femoropopliteal arteries in patients with subcritical or critical lower limb ischemia. MATERIALS AND METHODS: Ninety-two patients underwent 121 PTA procedures, 68 were of the superficial femoral artery (SFA), 13 of the popliteal and 40 of both arteries. Fifty-seven procedures were performed for treatment of occlusions. Eighty-four patients (94 procedures) were monitored with duplex scanning. RESULTS: Technical success rate was 88%. Primary success rates at 12 and 60 months in the whole series were 40% and 27%, respectively. The primary success rate in limbs with SFA occlusion of longer than 5 cm was only 12% after 5 years compared with 32% if the occlusion was .2) and was 89% and 89% for controls. The 5-year primary patency rates were 74% for octogenarians and 68% for septuagenarians (P >.2). Five-year limb salvage rates were 86% for octogenarians and 86% for septuagenarians. After a bypass graft, the respective rates of myocardial infarction were 4.1% and 3.9% per year and of a stroke 3.2% and 3.2% per year for octogenarians and septuagenarians, which occurred more frequently (P <.05) than in controls. CONCLUSIONS: Death and cardiovascular events are higher after revascularization in octogenarians and septuagenarians, compared with controls, and are related to the severity of atherosclerosis and not age. Patency rates are excellent and similar. Limb salvage procedures should be considered for most octogenarians. PMID- 11436087 TI - Use of cryopreserved cadaveric vein allograft for hemodialysis access precludes kidney transplantation because of allosensitization. AB - BACKGROUND: Dimethyl sulfoxide-cryopreserved cadaveric vein allografts have recently been proposed as an alternative to prosthetic grafts in the problem hemodialysis population. The transfer of mismatched major histocompatibility complex I and II molecules in association with these allografts can potentially lead to allosensitization in nonimmunosuppressed individuals. METHODS: In a university-affiliated medical center, 20 consecutive patients receiving technically successful upper arm cadaveric vein allograft fistulas (CAVFs) for hemodialysis between April 1999 and April 2000 were studied. A control cohort of 20 patients on a kidney transplantation waiting list was selected by nurses blinded to the study. These patients were matched for age, sex, history of transfusion, pregnancy, cause of kidney failure, and prior transplantation. The panel reactive antibody (PRA) values were recorded in this group over the same time period as the CAVF group. RESULTS: Patients receiving CAVFs had a mean PRA assay value of 84.1% (median, 96.5%) at an average of 3.1 months after engraftment (median, 1.5 months). The preengraftment PRA values were available for seven patients who were on the transplant waiting list. Six of these patients had nonreactive PRA assays before CAVF creation. All of these patients converted to positive PRA assays after CAVF creation with a mean value of 92.3% (median, 98%) at 2.85 months follow-up (median, 1.3 months). The mean PRA value for the control cohort was 5.5% (median, 2.5%), with no patients converting from a nonreactive to a reactive PRA assay during this same time interval. CONCLUSION: The use of dimethyl sulfoxide-cryopreserved cadaveric vein allografts for hemodialysis access leads to broad allosensitization as measured by PRA assay. Cryopreserved cadaveric vein allografts should not be used for hemodialysis access in potential kidney transplant recipients. PMID- 11436088 TI - Simultaneous analysis of 1176 gene products in normal human aorta and abdominal aortic aneurysms using a membrane-based complementary DNA expression array. AB - BACKGROUND: A number of changes in gene expression have been described in abdominal aortic aneurysms (AAAs), but the spectrum of molecular alterations in this disease is unknown. The purpose of this study was to characterize the expression of approximately 1000 gene products in human AAA tissue and to compare the profile of genes expressed in AAAs with that observed in normal aorta. MATERIALS AND METHODS: Total RNA was isolated from abdominal aortic wall tissues (4 AAAs and 4 normal aortas), and array-specific [(32)P]-labeled complementary DNA (cDNA) probes were created with reverse transcription. The cDNA probes were hybridized with nylon membranes containing an array of 1176 cDNA clones (AtlasArray Human 1.2 I; Clontech, Palo Alto, Calif), and autoradiographs were scanned to identify the patterns of gene expression characteristic of each tissue type. Densitometric analysis was used to standardize the expression of individual genes to a panel of housekeeping controls, and differential gene expression was defined by a signal ratio of at least 2:1. RESULTS: One hundred forty-five (12.3%) of the 1176 genes were consistently expressed in aortic tissue. Thymosin beta-4 was the most abundant of 101 transcripts detected in both AAAs and normal aorta, whereas 44 genes exhibited differential patterns of expression (39 predominant in AAAs and 5 in normal aorta). Densitometric analysis confirmed differences in expression for 20 of these gene products between AAAs and normal aorta, with the greatest increases seen for myeloid cell nuclear differentiation antigen (31-fold), cathepsin H (30-fold), platelet-derived growth factor-A (23 fold), apolipoprotein E (13-fold), gelatinase B/matrix metalloproteinase-9 (12 fold), and interleukin-8 (11-fold). The only gene products substantially decreased in AAAs were myosin light chain kinase (39-fold) and beta-1 integrin (twofold). AAA tissues thereby exhibited a distinct pattern of gene expression reflecting chronic inflammation, extracellular matrix degradation, atherosclerosis, and smooth muscle cell depletion. CONCLUSIONS: cDNA expression arrays provide a powerful new approach to help identify the molecular mechanisms responsible for aneurysmal degeneration. Further studies will be needed to elucidate the functional and pathophysiologic significance of the individual genes that exhibit altered levels of expression in AAA tissue. PMID- 11436089 TI - Topically applied tissue factor pathway inhibitor reduced intimal thickness of small arterial autografts in rabbits. AB - PURPOSE: The purpose of this study was to investigate whether topically applied tissue factor pathway inhibitor (TFPI) reduces intimal thickness and increases long-term patency of small arterial autografts in rabbits. METHODS: An entire 10 mm long section of the left femoral artery was harvested and immersed in saline solution (control group, n = 10), 100 IU/mL of heparin (heparin group, n = 15), or 40 microg/mL of TFPI (TFPI group, n = 15) for 15 minutes. Then the graft was interposed to the right femoral artery. Patency rates were determined by flow measurements throughout the time course of the study, and the grafts were analyzed for measurement of intimal thickness at 3 months after operation. Immunohistochemical analysis was performed to examine whether topically applied TFPI binds to endothelial cells of the grafts. RESULTS: Three-month postoperative patency rates were 10% in the control group, 47% in the heparin group, and 73% in the TFPI group. The TFPI group had a significantly higher patency rate than that of the control group (P <.005). Compared with the heparin group, the TFPI group had a significant reduction in intimal area (0.19 +/- 0.05 mm(2) vs 0.30 +/- 0.09 mm(2), P =.0051), in percentage of stenosis (35.7% +/- 7.7% vs 61.4% +/- 15.8%, P <.0001), and in intimal/media areas ratio (0.64 +/- 0.24 vs. 1.04 +/- 0.33, P =.0051). Immunohistologic analyses confirmed that topically applied TFPI bound to endothelial cells. CONCLUSION: These results indicate that topically applied TFPI reduces intimal thickness and increases long-term patency of small arterial autografts in rabbits. PMID- 11436090 TI - Adenovirus-mediated gene transfer of human inducible nitric oxide synthase in porcine vein grafts inhibits intimal hyperplasia. AB - OBJECTIVE: The aim of this study is to determine whether adenoviral inducible nitric oxide synthase (iNOS) gene transfer could inhibit intimal hyperplasia (IH) in porcine internal jugular veins interposed into the carotid artery circulation. METHODS: Porcine internal jugular veins were transduced passively with 1 x 10(11) particles of an adenoviral vector carrying either the human iNOS (AdiNOS) or beta galactosidase (AdlacZ) cDNA for 30 minutes and then interposed into the carotid artery circulation. Segments of each vein graft were maintained in an ex vivo organ culture to measure nitrite accumulation, a marker of nitric oxide synthesis. The grafts were analyzed immunohistochemically for the presence of neutrophils, macrophages, and leukocytes by staining for myeloperoxidase, ED1, and CD45, respectively, at 3 (n = 4) and 7 (n = 4) days. Morphometric analyses and cellular proliferation (Ki67 staining) were assessed at 3 (n = 4), 7 (n = 4), and 21 days (n = 8). RESULTS: AdlacZ-treated vein grafts demonstrated high levels of beta-galactosidase expression at 3 days with a gradual decline thereafter. Nitrite production from AdiNOS-treated vein grafts was approximately fivefold greater than AdlacZ-treated grafts (P =.00001). AdiNOS or AdlacZ treatment was associated with minimal graft inflammation. Cellular proliferation rates were significantly reduced in AdiNOS-treated grafts as compared with controls at both 3 (41%, P =.000004) and 7 days (32%, P =.0001) after bypass. This early antiproliferative effect was most pronounced at the distal anastomosis (65%, P =.0005). The iNOS gene transfer reduced the intimal/medial area ratio in vein grafts at 7 (36%, P =.009) and 21 days (30%, P =.007) versus controls. This inhibition of IH was again more prominent in the distal segments of the grafts (P =.01). CONCLUSION: Adenovirus-mediated iNOS gene transfer to porcine internal jugular vein grafts effectively reduced cellular proliferation and IH. Although iNOS gene transfer reduced IH throughout the entire vein graft, the most pronounced effect was measured at the distal anastomosis. These results suggest potential for iNOS-based genetic modification of vein grafts to prolong graft patency. PMID- 11436091 TI - Candida infection associated with a solitary mycotic common iliac artery aneurysm. AB - We report on a case of an isolated common iliac artery aneurysm infected by Candida albicans. To our knowledge, only one other case of this condition has been reported. The patient, a 49-year-old man with diabetes mellitus and a history of fungal urinary tract infections, had recurrent right knee pain and swelling. The knee effusion grew C albicans. Mild right hydronephrosis and a 4.6 cm aneurysm of the right common iliac artery without involvement of the aorta or iliac bifurcation was revealed by means of a computed tomography scan. The aneurysm wall was inflammatory, and there was associated purulence at the time of operation. The right ureter was densely adherent to the anterior aspect of the aneurysm, but could be palpated and dissected free because of a ureteral stent that was placed before the surgical incision. The aneurysm was resected, and the proximal and distal margins were oversewn without graft placement. C albicans was found in the resected aneurysm. The patient recovered without limb-threatening ischemia or claudication, but the distance he could walk remained limited because of right knee symptoms. The aneurysm may have formed by direct extension of infection from the right ureter or by hematogenous or lymphatic spread. This case raises interesting issues about operative strategies and etiology. PMID- 11436092 TI - Saccular renal artery aneurysm treated with an autologous vein-covered stent. AB - A 32-year-old woman underwent an autologous saphenous vein-covered stent deployment for the treatment of a saccular aneurysm on the distal renal artery. Complete exclusion of the aneurysm was immediately obtained. One year after the treatment, remarkable shrinkage of the aneurysm was demonstrated by means of computed tomography, and wide patency of the renal artery and the stent was shown by means of angiography. PMID- 11436093 TI - Vessel loops made easy. AB - Vessel loops are invaluable in vascular surgery. Unfortunately, they frequently slide toward the ends of the arteriotomy, making precise stitch placement difficult. By bringing the vessel loops out separate stab wounds, we improve visualization at the heel and toe of the anastomosis while preserving arterial elevation and hemostasis. PMID- 11436094 TI - Synopsis on cellular senescence and apoptosis. PMID- 11436095 TI - Regarding: "timing of postcarotid complications: a guide to safe discharge planning". PMID- 11436096 TI - The Food and Drug Administration approval of endovascular grafts for abdominal aortic aneurysm: an 18-month retrospective. PMID- 11436097 TI - Regarding "high prevalence of mild hyperhomocysteinemia in patients with abdominal aortic aneurysm". PMID- 11436099 TI - Engraftment syndrome following hematopoietic stem cell transplantation. AB - During neutrophil recovery following hematopoietic stem cell transplantation, a constellation of symptoms and signs including fever, erythrodermatous skin rash, and noncardiogenic pulmonary edema often occur. These clinical findings have usually been referred to as engraftment syndrome, or, reflecting the manifestations of increased capillary permeability, capillary leak syndrome. While described most often following autologous stem cell transplantation, a similar clinical syndrome has been observed followed allogeneic stem cell transplantation. Distinction from graft-versus-host disease in the allogeneic setting however, has been difficult. Recent experience with non-myeloablative conditioning for stem cell transplantation, however, reveals that an engraftment syndrome independent of GVHD may occur. In some cases, this engraftment syndrome may be a manifestation of a host-versus-graft reaction (graft rejection). While cellular and cytokine interactions are believed to be responsible for these clinical findings, a distinct effector cell population and cytokine profile have not been defined. Engraftment syndromes are likely associated with an increased transplant-related mortality, mostly from pulmonary and associated multi-organ failure. Corticosteroid therapy is often dramatically effective for engraftment syndrome, particularly for the treatment of the pulmonary manifestations. A proposal for a more uniform definition of engraftment syndrome has been developed in order to allow for a reproducible method of reporting of this complication and for evaluating prophylactic and therapeutic strategies. PMID- 11436100 TI - Hematopoietic stem cell transplantation activity in Europe 1999. AB - This survey on transplantation of hematopoietic stem cells from blood or bone marrow in Europe, the 10th in a series, reports the numbers of transplants performed in 1999 and concentrates on changes in indications and donor types. Members of the European Group for Blood and Marrow Transplantation and associated teams are invited every year to report their transplant numbers by indication, donor type and stem cell source. In 1999, a total 21 430 transplants were performed by 580 teams in 35 European countries. Of these transplants 18 720 were first transplants, 5879 (31%) allogeneic, 12 841 (69%) autologous; an additional 562 allogeneic and 2148 autologous transplants were re- or multiple transplants. Ninety-five percent of the autologous transplants and 45% of the allogeneic transplants were peripheral blood stem cell transplants. A total of 103, respectively 1.8% of the allogeneic transplants, were cord blood cell transplants. Main indications in 1999 were leukemias with 6289 transplants (34%), 70% thereof allogeneic transplants; lymphomas with 8219 transplants (44%), 92% thereof autologous transplants; solid tumors with 3302 transplants (18%), 99% thereof autologous transplants; nonmalignant disorders with 715 transplants (4%), 85% thereof allogeneic transplants. Absolute numbers of allogeneic transplants continued to increase as in previous years by 10%, in contrast, there was for the first time in 10 years a decrease in autologous transplants, mainly for solid tumors. Reasons therefore are discussed. These data reflect the most recent changes in utilisation and document current status of blood and marrow transplantation in Europe. PMID- 11436101 TI - Donating stimulated peripheral blood stem cells vs bone marrow: do donors experience the procedures differently? AB - As the demand for undifferentiated stem cells for the treatment of leukemia and other cancers has increased, new methods for their collection have been developed. One of these new methods, allogeneic peripheral blood stem cell (PBSC) donation, involves the administration of a granulocyte colony-stimulating factor (G-CSF, filgrastim), and a 1-2 day apheresis collection procedure. Our goal in the current study was to examine donors' psychosocial and physical experiences of PBSC vs marrow donation. Potential participants included 80 donors from the National Marrow Donor Program (NMDP) who donated a second time between 1991 and 1997. All of these donors had previously donated marrow. A final cohort of 70 donors (25 PBSC and 45 marrow) participated in a retrospective questionnaire study of their donation experiences. In general, all second-time donors reported low levels of concern about the physical consequences of donation. However, PBSC donors were more likely to have postponed the decision to donate a second time. Despite their reservations, PBSC donors reported fewer donation-related side effects than did marrow donors. Finally, PBSC donors reported that marrow donation was more physically difficult, time-consuming, and inconvenient, and that they preferred PBSC to marrow donation. PMID- 11436102 TI - High-dose melphalan with G-CSF-stimulated whole blood rescue followed by stem cell harvesting and busulphan/cyclophosphamide with autologous stem cell transplantation in multiple myeloma. AB - In 90 consecutive patients with multiple myeloma, we investigated the feasibility of administering a tandem high-dose therapy regimen, using whole blood for rescue after the first and leucapheresis harvested between the two high doses, for rescue after the second high dose. After 5 days of G-CSF 1 litre of whole blood (WB) was obtained, left undisturbed at 4 degrees C and reinfused 24 h after HDM (140 mg/m(2)). Patients not in progression after 3-6 months were again mobilised, leucapheresed and treated with busulphan 16 mg/kg and cyclophosphamide 120 mg/kg (Bu/Cy) and reinfusion. In 90 patients, WB contained a mean (range) of 0.57 (0.02 3.22) x 10(6)/kg CD34(+) cells. Recovery after HDM was in 13 days for granulocytes and in 18 days for platelets, with 11 patients not recovering within 3 months. There were three toxic deaths. Sixty-six patients qualified for harvesting after HDM. In the first 11, marrow was harvested. The subsequent 55 patients were mobilised and in 45 the preset minimum of 1.5 x 10(6) CD34(+) cells was obtained. Forty-nine patients actually received Bu/Cy. Recovery after Bu/Cy and marrow reinfusion was in 35 days for granulocytes and 20 days for platelets, with two of five patients not recovering after 3 months. After Bu/Cy and leucapheresis reinfusion, recovery was in 17 days for granulocytes and in 34 days for platelets. Nine patients did not recover within 3 months. There were four toxic deaths. The median overall survival from diagnosis for patients receiving HDM was 49 months and for patients also receiving Bu/Cy, 84 months. We conclude that WB rescue after HDM followed by leucapheresis and a second transplant is feasible in the majority of patients. Better mobilisation techniques are required to increase the number of patients who can receive the second transplant. PMID- 11436103 TI - High-dose chemotherapy (HDC) and autologous hematopoietic stem cell transplantation (ASCT) as salvage therapy for relapsed osteosarcoma. AB - In this report, we describe our experience with high-dose chemotherapy (HDC) and autologous stem cell transplantation (ASCT) in 15 children with relapsed osteosarcoma who were treated by members of the Cooperative Osteosarcoma Study Group. Eight patients received HDC after the first relapse, six patients after the second relapse and one after the sixth relapse. Thirteen patients underwent HDC and ASCT in complete remission and two patients had macroscopic tumor residues. Seven patients received HDC based on melphalan and etoposide. Four of these patients were treated with additional carboplatinum. Two patients received carboplatinum, etoposide, and thiotepa or cyclophosphamide. In six patients double HDC was performed. In all six of these, the first HDC consisted of thiotepa/ cyclophosphamide. The second regimens included melphalan/etposide (two patients), melphalan/etposide/ carboplatinum (one patient), and melphalan/busulfan (one patient). Three of the 15 patients died of toxic complications. Eight patients developed further relapses, two patients showed persistent disease, and two patients are presently in continuous complete remission. The probability of relapse-free survival was 0.20 +/- 0.12 within a median follow-up (MFU) of 8 months and the probability of overall survival was 0.29 +/- 0.12 after a MFU of 16 months. In conclusion, utilization of HDC and ASCT in this patient group did not significantly improve the treatment outcome compared to conventional relapse therapy. PMID- 11436104 TI - Double dose-intensive chemotherapy with autologous stem cell support for relapsed and refractory testicular cancer: the University of Michigan experience and literature review. AB - Testicular cancer patients refractory or in relapse after primary chemotherapy have < or =25% 5-year progression-free survival with salvage. To improve prognosis, patients entered a phase I/II tandem dose-escalation trial of carboplatin (1500-2100 mg/m(2)) and etoposide (1200-2250 mg/m(2)) with ABMT. Patients were eligible for a second cycle if disease progression was absent and performance status allowed. From August 1990 to June 1998, 29 males (25 NSGCT) were treated. At the time of ABMT, 10 were chemosensitive, four were chemoresistant, and 10 were absolutely refractory to platinum. Disease status (no. patients) at transplant: primary refractory disease (six), first relapse (10), second relapse (eight), third relapse (five). Fifteen (52%) received both transplants. Treatment-related mortality was 10%. Best response after ABMT included: two CR, one CR surgically NED, five PR, three PR surgically NED, seven SD, and eight PD. Eight (28%) patients are continuously progression-free a median 60 months (range, 31-93) from first ABMT. Three seminoma patients remain progression-free. Of five long-term NSGCT survivors, four were treated in first relapse with platinum-sensitive disease. Eighteen relapses occurred a median of 4 months after ABMT I (two late relapses at 28 and 44 months). The median PFS and OS for the whole group are 4 and 14 months, respectively. Patients with relapsed/ refractory testicular cancer benefit most from ABMT if they have platinum sensitive disease in first relapse. Patients who do poorly despite ABMT have a mediastinal primary site, true cisplatin-refractory disease, disease progression prior to ABMT, and/or markedly elevated betaHCG at ABMT. New treatment modalities are needed for the latter group. PMID- 11436105 TI - The predictive value of helper T lymphocyte precursor frequencies for graft versus-host disease and graft-versus-leukaemia effects in allogeneic bone marrow transplantation. AB - Helper T lymphocyte precursor (HTLp) frequencies determined by limiting dilution analysis were studied in the graft-versus-host direction to assess the predictive value for outcome in allogeneic BMT. The HTLp frequencies correlated with the degree of HLA disparity. HTLp frequencies from 28 HLA-identical sibling BMT pairs had a median of 1:557 362 (range 1:9511 to <1:2 500 000). The HTLp frequencies from 20 HLA-matched unrelated and partially HLA-matched related BMT pairs had a median of 1:88 110 (range 1:4139-1:736 123). The HLA-identical sibling BMT pairs were split evenly into high and low HTLp frequency groups above and below 1:500 000. There was a trend towards a higher risk for acute GVHD > or =grade II (P = 0.075) in the high frequency group. There was no difference in TRM. The high HTLp frequency group had a significantly higher risk for chronic GVHD (P = 0.04), a significantly lower risk for relapse (P = 0.01), as well as a significantly better overall survival (P = 0.045) and leukaemia-free survival (P = 0.008). The HLA-matched unrelated and partially HLA-matched related BMT pairs were split evenly into high and low HTLp frequency groups above and below 1:90 000. There was a significantly higher risk for acute GVHD > or = grade II (P = 0.007) in the high HTLp frequency group. There was a trend towards a higher TRM in the high HTLp frequency group (P = 0.05). There were no differences in chronic GVHD, risk of relapse, overall survival and leukaemia-free survival. Analyzing all 48 patients the risk of acute GVHD > or = grade II and TRM was significantly higher (P = 0.012 and 0.021, respectively) with HTLp frequencies >1:100 000 and there was a trend towards a higher risk of relapse (P = 0.058) with low HTLp frequencies <1:400 000. Patients in the intermediate HTLp frequency group 1:100 000-1:400 000 had a trend towards improved survival (P = 0.059). The HTLp frequency seems to detect clinically significant differences in alloreactivity, that can be useful in donor selection and graft engineering. PMID- 11436106 TI - Long-term haematological recovery following high-dose chemotherapy with autologous bone marrow transplantation or peripheral stem cell transplantation in patients with solid tumours. AB - long-term peripheral blood counts and factors influencing long-term trilineage haematological recovery of consecutive patients in a single institution treated with high-dose chemotherapy (hdc) and abmt or psct for solid tumours were examined. patients with a relapse-free survival of >1 year were included in the analysis (n = 131). Peripheral blood counts were examined 6 months and yearly following transplantation. Median follow-up was 4.1 years (range 1-10+ years). Three years after transplantation 91% of patients had normal white blood counts (WBC), 94% normal haemoglobin (Hb) and 75% normal platelets. Trilineage recovery was complete in 70% (n = 83) at 3 years and 85% (n = 50) at 5 years. Recovery of Hb occurred before WBC and platelet recovery. Approximately 25% of patients displayed an elevated MCV throughout the follow-up period. These long-term results were independent of age, high-dose regimen, number of reinfused stem cells and stem cell source. Double (n = 12) vs single (n = 119) transplantations showed significantly slower trilineage recovery and higher MCV. No secondary graft failure, myelodysplasia or leukaemia was encountered. In conclusion, complete trilineage recovery after HDC followed by ABMT or PSCT occurs slowly. PSCT and ABMT are capable of maintaining long-term haematopoiesis. Slower recovery is seen after double transplantations. The results suggest lasting implications for bone marrow function after autologous transplantation. PMID- 11436107 TI - Use of first line bronchoalveolar lavage in the immunosuppressed oncology patient. AB - Immunosuppressed oncology patients who develop pulmonary infiltrates during treatment have a mortality rate of the order of 55-90%. Early diagnosis and treatment is associated with increased survival. At present, diagnosis relies on invasive sampling of the respiratory tract using fibre-optic bronchoscopy. We have looked at a 30-month period, from June 1997 to December 1999, where 25 bronchoscopies were performed on patients from the Lymphoma and BMT units at The Royal Marsden Hospital for the further investigation of pulmonary infiltrates. Nine bronchoscopies (36%) yielded a positive result and seven (28%) led to a change in management. Analysis of the data showed that neither a positive result nor a change in management had any impact on overall survival. After reviewing the background literature on the investigation of pulmonary infiltrates in this group and discussion of the respective merits and limitations, we propose a management flowchart, with high-resolution computed tomography (HRCT) as the test arm in a future randomised trial of these patients. PMID- 11436108 TI - Tuberculosis among allogeneic bone marrow transplant recipients in India. AB - Allogeneic bone marrow transplant recipients have severe impairment of cell mediated immunity and hence a higher incidence of mycobacterial infections might be expected in regions where tuberculosis is common. We reviewed the case records of 217 patients who underwent allogeneic bone marrow transplantation during the period 1986-1999 at our center in India. Mycobacterial infections were diagnosed in three patients (1.38%). All patients presented with extrapulmonary disease. Two patients had disseminated tuberculosis with one of these being diagnosed on autopsy studies. The third patient had tuberculosis involving the cervical lymph node and dorsal spine. Two patients treated with antituberculous therapy are well. Infection with Mycobacterium tuberculosis is not a common problem in allogeneic bone marrow recipients even in an endemic area, but when it occurs, it is usually disseminated with predominantly extrapulmonary involvement. PMID- 11436109 TI - Serum levels of soluble adhesion molecules in stem cell transplantation-related complications. AB - To assess the involvement of vascular endothelial cell activation and damage in stem cell transplantation (SCT)-related complications, such as acute and chronic GVHD and thrombotic microangiopathy (TMA), we investigated the changes in serum levels of soluble forms of vascular cell adhesion molecule-1 (sVCAM-1) and E selectin (sE-selectin) in SCT. The soluble form of intercellular adhesion molecule-1 (sICAM-1) was also analyzed. In patients with acute GVHD (grades II IV), serum levels of sE-selectin and sICAM-1 increased around onset of GVHD (day 30). While the increase of sE-selectin levels was transient, sICAM-1 levels remained high until day 60. In patients with extensive chronic GVHD, sVCAM-1 as well as sE-selectin levels significantly increased. The appearance of clinical symptoms was preceded by elevations of sVCAM-1 and sE-selectin levels on day 60, and sICAM-1 levels on days 30 and 60. For the analysis of TMA, to exclude the influence of GVHD, serum levels were measured in auto-SCT patients. Around the onset of TMA, sVCAM-1 and sE-selectin levels significantly increased in patients with TMA without an increase of sICAM-1 levels. These findings support the notion that activation and injury of endothelium are commonly involved in the pathogenesis of acute and chronic GVHD and TMA. PMID- 11436110 TI - Marked reduction in the incidence of hepatic veno-occlusive disease after allogeneic hematopoietic stem cell transplantation with CD34(+) positive selection. AB - Veno-occlusive disease of the liver (VOD) is a common and severe complication of allogeneic hematopoietic stem cell transplantation (HSCT). To determine the incidence of, and the risk factors for the development of VOD, we performed a retrospective analysis of a series of 178 patients, who underwent allogeneic HSCT at our institution between 1990 and 1999. Busulfan and cyclophosphamide constituted the conditioning regimen most frequently administered. Bone marrow was the source of stem cells in 129 patients (73%), and peripheral blood (PBSC) in 49 patients (27%). Thirty-one patients of the PBSC group received CD34(+) positively selected grafts. Most patients were given cyclosporin A and methotrexate (MTX) as graft-versus-host disease (GVHD) prophylaxis. Overall, 30 patients (17%) developed VOD. In univariate analyses, the incidence of VOD was significantly higher in recipients of unmanipulated grafts (20% vs 0%; P = 0.01), in patients with active malignant disease at transplantation (24% vs 9%; P = 0.03), in recipients of marrow from unrelated donors (33% vs 15%; P = 0.03), in patients grafted with bone marrow (21% vs 6%; P = 0.03), and in those receiving MTX as GVHD prophylaxis (21% vs 6%; P = 0.05). Under multivariate analysis, only CD34(+) positive selection (P = 0.0004) and the status of the disease at transplant (P = 0.03) were statistically significant variables for the development of VOD. We conclude that CD34(+) positively selected PBSC transplantation could result in a marked reduction in the incidence of VOD after allogeneic HSCT. PMID- 11436111 TI - Longitudinal effects of high-dose chemotherapy and autologous stem cell transplantation on quality of life in the treatment of metastatic breast cancer. AB - This study determined the effects of high-dose chemotherapy (HDCT) with autologous blood stem cell transplantation (ASCT) on quality of life (QL) in women with metastatic breast cancer prior to, and during treatment, and up to 1 year post-ASCT. Thirty-three women diagnosed with metastatic breast cancer participated in a phase 1 clinical trial of a new combination of cyclophosphamide (CTX) and mitoxantrone (MXT), with dose escalation of paclitaxel. Longitudinal QL data were collected using the functional living index-cancer (FLIC) and symptom scales at seven time periods: pre-induction chemotherapy (CT), post-induction CT, post-high dose CT (HDCT), and at 3, 6, 9 and 12 months post-ASCT. FLIC scores indicated that the worst problems for patients were feelings of hardship on themselves and their families, followed by psychological functioning and physical functioning problems. The time around diagnosis of the metastatic disease and following HDCT were the worst times for all levels of quality of life, but anxiety and depression symptoms continued to increase in severity across the entire follow-up period. The symptoms that were most problematic were worry about the future, loss of sexual interest, anxiety about the treatment, general worrying, and joint pain. These data highlight the problems that women with metastatic breast cancer encounter at different stages of the disease and treatment process, and can be used to tailor psychosocial interventions appropriate for treating the relevant issues at different points in time. PMID- 11436112 TI - Effect of induced GVHD in leukemia patients relapsing after allogeneic bone marrow transplantation: single-center experience of 33 adult patients. AB - In a retrospective single center study, we examined the outcome of induced GVHD in leukemia patients relapsing after allogeneic BMT. Thirty-three adult patients with leukemia (15 AML, 3 ALL, and 15 CML) persisting or relapsing 1-36 months (median, 6) after allogeneic BMT underwent various immune manipulations and consequently developed acute and/or chronic GVHD at our center. Immunotherapies to elicit GVHD comprised chemotherapy followed by PBSC (n = 18), non myeloablative transplant (n = 2), PBL followed by IFN-alpha (n = 5), PBL alone (n = 3), abrupt cessation of CsA (n = 3), and CsA withdrawal combined with IFN-alpha (n = 2). Twenty-four (72.7%) patients obtained a remission including complete hematological or cytogenetic remission, respectively, for acute leukemias or CML. Overall survival of patients, estimated at 3 years using the Kaplan-Meier method, was 33.9% (95% CI, 20-52%). Twelve patients including two patients with ALL remain in complete hematological (n = 5) or cytogenetic remission (n = 7) 3-93 months (median 12) after achieving remission. Twelve (63.2%) of 19 dead patients died due to treatment-related toxicities; five patients from acute GVHD, three from GVHD followed by infections and four from infections. By multivariate Cox analysis, only chronic GVHD resulted in a higher probability of disease-free survival (P = 0.026). Eight patients who had both acute GVHD < or = grade I and chronic GVHD are all alive without leukemia. We conclude that acute GVHD is associated with considerable toxicity while chronic GVHD plays a role in retaining remission in leukemia relapsing after allogeneic BMT. PMID- 11436113 TI - Hepatic graft-versus-host disease presenting as an acute hepatitis after allogeneic peripheral blood stem cell transplantation. AB - Hepatic graft-versus-host disease (GVHD) generally presents as cholestatic jaundice, and increased serum alkaline phosphatase (ALP) is followed by hyperbilirubinemia and clinical jaundice. Currently accepted standards for evaluating the clinical severity of GVHD are based not on serum aminotransferase levels but on the serum bilirubin level. We describe a 17-year-old Japanese female who had increased aminotransferases without cholestasis on day 23 after allogeneic peripheral blood stem cell transplantation (allo-PBSCT). Liver biopsy revealed lymphocytic infiltration of the portal tracts and pericentral necrosis of the lobuli. The limiting plates were not clearly defined due to cellular infiltrates. There was periductal lymphocytic infiltration and vacuolization of the biliary epithelial cells with exocytosis, compatible with GVHD of cholangiohepatitic type. These findings indicate that acute hepatic GVHD may present as acute hepatitis and this should be included in the differential diagnosis for patients with increased aminotransferases after allogeneic stem cell transplantation. PMID- 11436115 TI - Mucositis associated with stem cell transplantation: current status and innovative approaches to management. AB - Treatment-related morbidity, and in some cases, mortality, associated with autologous and allogeneic bone marrow transplantation has decreased in the past decade largely due to the use of blood stem cells combined with hematopoietic growth factors. However, these procedures remain morbid, with several series documenting regimen-related injury to the oral mucous membranes, the worst form of toxicity from a patient perspective. The pathophysiology of transplant-related mucositis is related to two major events: direct mucosal basal cell injury leading to atrophy and ulcerations, and local infections that can become systemic, the latter of which are exacerbated by the severe neutropenia accompanying high-dose chemotherapy. Recent investigational agents designed to interfere with these two aspects of mucositis have been developed and are showing promise in early clinical trials. In particular, keratinocyte growth factor (KGF) and interleukin-11 appear active. They increase basal cell proliferation, prevent apoptosis due to the preparative regimen, and appear to ameliorate the mucositis seen with high-dose chemotherapy regimens. Oral, nonabsorbable anti-infective agents are also being tested in an attempt to prevent both local and systemic infections. Devoid of significant side-effects, KGF is now in large phase 2 trials that, if positive, will be a significant advance in promoting less morbid transplants by reducing pain and the risk of secondary infections and thus may reduce supportive care costs. PMID- 11436116 TI - Nonmyeloablative preparative regimens for allogeneic hematopoietic transplantation. AB - Allogeneic hematopoietic transplantation is an effective therapy for a range of malignancies. High doses of myelosuppressive chemotherapy or radiation have been used in preparative regimens with the goal of preventing graft rejection and eradicating malignancy. Much of the benefit of transplantation, however, results from graft-versus-malignancy effects, mediated by donor immunocompetent cells. An alternative approach is to utilize less toxic, nonmyeloablative preparative regimens to achieve engraftment and allow graft-versus-malignancy effects to develop. This strategy reduces the risk of treatment-related mortality and allows transplantation for elderly or medically infirm patients not eligible for myeloablative therapy. Nonmyeloablative preparative regimens appear promising in diagnoses sensitive to graft-versus-malignancy effects and provide a platform for further development of cellular immunotherapy. Controlled clinical trials are warranted to define the role of nonmyeloablative allogeneic transplants in a range of hematologic malignancies and selected solid tumors. PMID- 11436117 TI - Mobilization of peripheral blood stem cells following myelosuppressive chemotherapy: a randomized comparison of filgrastim, sargramostim, or sequential sargramostim and filgrastim. AB - Myelosuppressive chemotherapy is frequently used for mobilization of autologous CD34(+) progenitor cells into the peripheral blood for subsequent collection and support of high-dose chemotherapy. The administration of myelosuppressive chemotherapy is typically followed by a myeloid growth factor and is associated with variable CD34 cell yields and morbidity. The two most commonly used myeloid growth factors for facilitation of CD34 cell harvests are granulocyte colony stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF). We performed a randomized phase III clinical trial comparing G-CSF, GM CSF, and sequential administration of GM-CSF and G-CSF following administration of myelosuppressive chemotherapy. We evaluated CD34 yields, morbidity, and cost effectiveness of the three cytokine schedules. One hundred and fifty-six patients with multiple myeloma, breast cancer, or lymphoma received cyclophosphamide with either paclitaxel or etoposide and were randomized to receive G-CSF 6 microg/kg/day s.c., GM-CSF 250 microg/m(2)/day s.c., or GM-CSF for 6 days followed by G-CSF until completion of the stem cell harvest. Compared with patients who received GM-CSF, patients who received G-CSF had faster recovery of absolute neutrophil count to 0.5 x 10(9) per liter (median of 11 vs14 days, P = 0.0001) with fewer patients requiring red blood cell transfusions (P= 0.008); fewer patients with fever (18% vs 52%, P = 0.001); fewer hospital admissions (20% vs 42%, P = 0.13); and less intravenous antibiotic therapy (24% vs 59%, P = 0.001). Patients who received G-CSF also yielded more CD34 cells (median 7.1 vs 2.0 x 10(6) kg per apheresis, P = 0.0001) and a higher percentage achieved 2.5 x 10(6) CD34 cells per kilogram (94% vs 78%, P = 0.21) and 5 x 10(6) CD34 cells per kilogram (88% vs 53%, P = 0.01) or more CD34 cells per kilogram with fewer aphereses (median 2 vs 3, P = 0.002) and fewer days of growth factor treatment (median 12 vs 14, P = 0.0001). There were no significant differences in outcomes between groups receiving G-CSF alone and the sequential regimen. After high-dose chemotherapy, patients who had peripheral blood stem cells mobilized with G-CSF or the sequential regimen received higher numbers of CD34 cells and had faster platelet recovery with fewer patients requiring platelet transfusions than patients receiving peripheral blood stem cells mobilized by GM-CSF. In summary, G CSF alone is superior to GM-CSF alone for the mobilization of CD34(+) cells and reduction of toxicities following myelosuppressive chemotherapy. An economic analysis evaluating the cost-effectiveness of these three effective schedules is ongoing at the time of this writing. PMID- 11436119 TI - Meiotic segregation analysis in a t(4;8) carrier: comparison of FISH methods on sperm chromosome metaphases and interphase sperm nuclei. AB - Meiotic segregation of a t(4;8)(q28;p23) translocation carrier was determined by two different methods to compare the final results. A total of 352 sperm chromosome complements, obtained after human-hamster in vitro fertilisation, were analysed by whole chromosome painting, and 6590 sperm heads were studied by fluorescence in situ hybridisation (FISH). Frequencies of alternate, adjacent I, adjacent II and 3 : 1 segregations were, for sperm chromosomes, 35.5, 33.2, 19.9 and 11.3% respectively. For sperm head analysis, results were 30.5, 28.5, 20.5 and 19.5% respectively. There were no statistically significant differences between the two methods with respect to the observed frequencies of sperm with balanced and unbalanced chromosome constitutions. Among unbalanced gametes, the methods differed only in the frequency of 3 : 1 segregation (chi(2), P<0.0001). Different factors that could explain this result are discussed. To determine possible interchromosomal effects, multicolour FISH was used on sperm heads. Disomy rates of sex and 18 chromosomes were higher in the translocation carrier than in the control. The differences observed were statistically significant (P<0.0001 for chromosomes X and 18, and P=0.0091 for chromosome Y). PMID- 11436120 TI - Evaluation of first trimester maternal serum and ultrasound screening for Down's syndrome in Eastern and Northern Finland. AB - The current trend in prenatal diagnosis is that trisomy screening is being moved to the first trimester and ultrasonographic nuchal translucency measurement is included in risk calculation. It is likely that biochemical screening in the second trimester will gradually be given up. In Eastern and Northern Finland, during the year 1999 we offered first-trimester ultrasonographic and serum screening for trisomy 21, with measurements of maternal serum PAPP-A and beta hCG. A total of 2515 pregnant women participated in the screening, yielding the detection of eight foetuses with Down's syndrome. Six affected foetuses (75%) were detected by means of first-trimester serum screening. Since we were in the phase of collecting data for the Finnish medians for PAPP-A and beta-hCG, the women were not given the estimates of risk for trisomy 21. Only 1602 of the 2515 enrolled women had the combination of first-trimester ultrasonographic and serum screening performed, and in that group there were five foetuses with Down's syndrome. The combination ultrasonographic and serum approach yielded a Down's syndrome detection rate of 80% (four out of five) with a 5% false positive rate, whereas in nuchal translucency based-screening the detection rate was 60%, with a 5% false positive rate. PMID- 11436121 TI - Analysis of the methylation status of the KCNQ1OT and H19 genes in leukocyte DNA for the diagnosis and prognosis of Beckwith-Wiedemann syndrome. AB - Beckwith-Wiedemann syndrome (BWS) is an overgrowth disorder involving developmental abnormalities, tissue and organ hyperplasia and an increased risk of embryonal tumours (most commonly Wilms tumour). This multigenic disorder is caused by dysregulation of the expression of imprinted genes in the 11p15 chromosomal region. Molecular diagnosis of BWS is currently difficult, mostly due to the large spectrum of genetic and epigenetic abnormalities. The other difficulty in managing BWS is the identification of patients at risk of tumour. An imprinted antisense transcript within KCNQ1, called KCNQ1OT (also known as LIT1), was recently shown to be normally expressed from the paternal allele. A loss of imprinting of the KCNQ1OT gene, associated with the loss of maternal allele-specific methylation of the differentially methylated region KvDMR1 has been described in BWS patients. The principal aim of this study was to evaluate the usefulness of KvDMR1 methylation analysis of leukocyte DNA for the diagnosis of BWS. The allelic status of the 11p15 region and the methylation status of the KCNQ1OT and H19 genes were investigated in leukocyte DNA from 97 patients referred for BWS and classified into two groups according to clinical data: complete BWS (CBWS) (n=61) and incomplete BWS (IBWS) (n=36). Fifty-eight (60%) patients (39/61 CBWS and 19/36 IBWS) displayed abnormal demethylation of KvDMR1. In 11 of the 56 informative cases, demethylation of KvDMR1 was related to 11p15 uniparental disomy (UPD) (nine CBWS and two IBWS). Thirteen of the 39 patients with normal methylation of KvDMR1 displayed hypermethylation of the H19 gene. These 13 patients included two siblings with 11p15 trisomy. These results show that analysis of the methylation status of KvDMR1 and the H19 gene in leukocyte DNA is useful in the diagnosis of 11p15-related overgrowth syndromes, resulting in the diagnosis of BWS in more than 70% of investigated patients. We also evaluated clinical and molecular features as prognostic factors for tumour and showed that mosaicism for 11p15 UPD and hypermethylation of the H19 gene in blood cells were associated with an increased risk of tumour. PMID- 11436122 TI - Novel RET mutations in Hirschsprung's disease patients from the diverse South African population. AB - Hirschsprung's disease (HSCR) is a common cause of intestinal obstruction in neonates with an incidence of one in 5000 live births. The disease occurs due to the absence of parasympathetic neuronal ganglia in the hindgut, resulting in irregular or sustained contraction of the affected segment. DNA samples of 40 unrelated subjects with HSCR were subjected to mutation screening of the RET (REarranged during Transfection) proto-oncogene, the major susceptibility gene for HSCR. Five novel (V202M, E480K, IVS10-2A/G, D771N, IVS19-9C/T) and one previously described mutation (P973L) were identified. Only two of the mutation positive patients (from different ethnic groups) displayed total colonic aganglionosis, and both were heterozygous for mutation D771N. The potential disease-causing mutations occurred in 20% of individuals, with more males (22.5% representing seven of 31 males) affected than females (12.5% representing one of eight females). This study represents the first comprehensive genetic analysis of this disease in the diverse South African population. PMID- 11436123 TI - BRCA1 and BRCA2 mutations among 233 unselected Finnish ovarian carcinoma patients. AB - Germline mutations of BRCA1 and BRCA2 predispose to hereditary breast-ovarian cancer syndrome. In Finland, 20 different BRCA1/2 mutations have been identified, and 13 of them are founder mutations that account for the vast majority of Finnish BRCA1/2 families. The purpose of our study was to determine the prevalence of BRCA1/2 mutations in unselected Finnish ovarian carcinoma patients and to evaluate the relationship between mutation carrier status and personal/family history of cancer. Two hundred and thirty-three patients were screened for all the 20 BRCA1/2 mutations known in the Finnish population. Additionally, a subgroup of patients with personal history of breast cancer and/or family history of breast and/or ovarian cancer was screened for novel BRCA1/2 mutations. Thirteen patients (5.6%) had mutations: eleven in BRCA1 and two in BRCA2. All the mutation-positive patients were carriers of the previously known Finnish BRCA1/2 mutations, and seven recurrent founder mutations accounted for 12 of the 13 mutations detected. A logistic regression analysis was used to determine the odds of mutation for ovarian carcinoma patients. The most significant predictor of a mutation was the presence of both breast and ovarian cancer in the same woman, but family history of breast cancer was also strongly related to mutation carrier status. Although BRCA1/2 mutation testing is not warranted in the general Finnish ovarian cancer patient population, patients who have also been diagnosed with breast cancer or have family history of breast or breast and ovarian cancer could benefit from referral to genetic counselling and mutation testing. PMID- 11436124 TI - Multiple founder effects in spinal and bulbar muscular atrophy (SBMA, Kennedy disease) around the world. AB - SBMA (spinal and bulbar muscular atrophy), also called Kennedy disease, is an X chromosomal recessive adult-onset neurodegenerative disorder caused by death of the spinal and bulbar motor neurones and dorsal root ganglia. Patients may also show signs of partial androgen insensitivity. SBMA is caused by a CAG repeat expansion in the first exon of the androgen receptor (AR) gene on the X chromosome. Our previous study suggested that all the Nordic patients with SBMA originated from an ancient Nordic founder mutation, but the new intragenic SNP marker ARd12 revealed that the Danish patients derive their disease chromosome from another ancestor. In search of relationships between patients from different countries, we haplotyped altogether 123 SBMA families from different parts of the world for two intragenic markers and 16 microsatellites spanning 25 cM around the AR gene. The fact that different SBMA founder haplotypes were found in patients from around the world implies that the CAG repeat expansion mutation has not been a unique event. No expansion-prone haplotype could be detected. Trinucleotide diseases often show correlation between the repeat length and the severity and earlier onset of the disease. The longer the repeat, the more severe the symptoms are and the onset of the disease is earlier. A negative correlation between the CAG repeat length and the age of onset was found in the 95 SBMA patients with defined ages at onset. PMID- 11436125 TI - Lack of replication of association findings in complex disease: an analysis of 15 polymorphisms in prior candidate genes for sporadic Alzheimer's disease. AB - There is considerable enthusiasm for the prospect of using common polymorphisms (primarily single nucleotide polymorphisms; SNPs) in candidate genes to unravel the genetics of complex disease. This approach has generated a number of findings of loci which are significantly associated with sporadic Alzheimer's disease (AD). In the present study, a total of 15 genes of interest were chosen from among the previously published reports of significant association in AD. Genotyping was performed on polymorphisms within those genes (14 SNPs and one deletion) using Dynamic Allele Specific Hybridization (DASH) in 204 Swedish patients with sporadic late-onset AD and 186 Swedish control subjects. The genes chosen for analysis were; low-density lipoprotein receptor-related protein (LRP1), angiotensin converting enzyme (DCP1), alpha-2-macroglobulin (A2M), bleomycin hydrolase (BLMH), dihydrolipoyl S-succinyltransferase (DLST), tumour necrosis factor receptor superfamily member 6 (TNFRSF6), nitric oxide synthase (NOS3), presenilin 1 (PSEN1), presenilin 2 (PSEN2), butyrylcholinesterase (BCHE), Fe65 (APBB1), oestrogen receptor alpha (ESR1), cathepsin D (CTSD), methylenetetrahydrofolate reductase (MTHFR), and interleukin 1A (IL1A). We found no strong evidence of association for any of these loci with AD in this population. While the possibility exists that the genes analysed are involved in AD (ie they have weak effects and/or are population specific), results reinforce the need for extensive replication studies if we are to be successful in defining true risk factors in complex diseases. PMID- 11436126 TI - Haemochromatosis gene mutations in a clustered Italian population: evidence of high prevalence in people of Celtic ancestry. AB - Hereditary haemochromatosis is an inherited disorder characterised by an excessive iron absorption from the diet and is associated with several HFE gene mutations. One hypothesis is that these genetic mutations originated in the Celtic populations. The aim of this study is to determine the frequency of HFE gene mutations in a clustered Italian population of Celtic ancestry (Cimbri, Asiago plateau). One hundred and forty-nine consecutive unrelated blood donors (31 females and 118 males) were enrolled in this study. A family investigation was performed in each case to identify the ethnic origin of the individuals. The analysis of HFE gene mutations was performed by PCR amplification followed by digestion with RsaI and DpnII restriction enzymes. At least one HFE gene mutation was identified in 49 individuals (32.9%) of the studied population. The allele frequencies of the C282Y and H63D were respectively 0.037 and 0.144. When we considered only the 103 individuals with relatives born in Asiago, the prevalence of the HFE mutations rose from 32.9 to 39.8%; the allele frequencies of the C282Y and H63D were respectively 0.048 and 0.174. The mean serum iron and ferritin levels were significantly higher in individuals with the HFE mutations than in normal cases. This study indicates that the prevalence of the HFE gene mutations is surprisingly high in Italians with Celtic ancestry. This could suggest the need to perform large mass studies in selected areas of the country to detect the affected patients and prevent the disease in homozygous individuals. PMID- 11436127 TI - Genetic linkage of autosomal dominant primary open angle glaucoma to chromosome 3q in a Greek pedigree. AB - A locus for juvenile onset open angle glaucoma (OAG) has been assigned to chromosome 1q in families with autosomal dominant inheritance (GLC1A), due to mutations in the TIGR/MYOC gene. For adult onset OAG, called primary open angle glaucoma or POAG, five loci have so far been mapped to different chromosomes (GLC1B-GLC1F). Except for the GLC1B locus, the other POAG loci have so far been reported only in single large pedigrees. We studied a large family identified in Epirus, Greece, segregating POAG in an autosomal dominant fashion. Clinical findings included increased cup to disc ratio (mean 0.7), characteristic glaucomatous changes in the visual field, and intraocular pressure before treatment more than 21 mmHg (mean 31 mmHg), with age at diagnosis 33 years and older. Linkage analysis was performed between the disease phenotype and microsatellite DNA polymorphisms. Linkage was established with a group of DNA markers located on chromosome 3q, where the GLC1C locus has previously been described in one large Oregon pedigree. A maximal multipoint lod score of 3.88 was obtained at marker D3S1763 (penetrance 80%). This represents the second POAG family linked to the GLC1C locus on chromosome 3q, and haplotype analysis in the two families suggests an independent origin of the genetic defect. PMID- 11436128 TI - Linkage analysis in multiple sclerosis of chromosomal regions syntenic to experimental autoimmune disease loci. AB - Multiple sclerosis is a demyelinating disorder of the central nervous system with a putative autoimmune aetiology in which several genes are thought to be involved. Four published genomic screens have confirmed that a gene influencing MS resides within or close to the HLA class II region in 6p21. Still, this locus is likely to confer only a part of the genetic susceptibility in MS. Further, all four studies identified a number of other regions with possible linkage. We have investigated eight chromosomal intervals syntenic to loci of importance for experimental autoimmune model diseases in the rat in 74 Swedish MS families. Possible linkage (a non-parametric linkage NPL score of 1.16 by GENEHUNTER computer package) was observed with markers in 12p13.3, a region syntenic to the rat Oia2 locus which is importance for oil induced arthritis (OIA). Four markers in the T cell receptor beta chain gene region in 7q35 showed possible linkage (highest NPL score of 1.16). This locus is syntenic to the rat Cia3 locus (collagen induced arthritis). These two loci at least partially overlap with chromosomal regions showing indicative evidence for linkage in the previous MS genomic screens. Indeed, both Oia2 and Cia3 were recently found to be linked also with experimental autoimmune encephalomyelitis, a commonly used model for MS. Markers in 2p12, 3p25, 10q11.23, 17q21-25, 19q13.1, and 22q12-13 failed to provide evidence for linkage. We conclude that evidence is amounting that 12p13 12 and 7q34-36 may harbour genes with an importance for MS. The synteny with experimental loci may eventually facilitate their identification. PMID- 11436129 TI - Use of haplotype information to test involvement of the LRP gene in Alzheimer's disease in the French population. AB - The low density lipoprotein receptor-related protein gene (LRP) is a good candidate gene for Alzheimer's Disease (AD). Its protein is involved in the physiopathology of AD and has been found in senile plaques; on the other hand, LRP is located in 12q, a region in which genetic linkage to AD was reported. Two common polymorphisms, a tetranucleotide repeat in the 5' untranslated region and a single nucleotide polymorphism at position 766 in exon 3, were found to be associated with AD, but contradictory results were obtained in subsequent association studies. In the absence of clear hypotheses concerning the association of these polymorphisms with AD and their functional role, our objective was to test the association between AD and the two LRP polymorphisms in a large French case-control sample (274 Caucasian AD patients and 290 matched controls) using haplotype analysis. First, the separate study of each polymorphism showed no significant difference in genotype and allele frequencies between AD cases and controls. Second, strong linkage disequilibrium was found between alleles of the two polymorphisms in controls and in cases and the linkage disequilibrium between the 91 bp and C alleles were opposite in cases and in controls. Third, we found that the frequency of the 91-C haplotype was higher in cases than in controls, but the type I error was 0.061, slightly higher than the conventional one of 5%. The haplotype frequencies did not vary significantly as a function of age and APOE epsilon4 status. One interest in this study is the use of the haplotype analysis, which can be used to combine information from several polymorphisms, taking into account their dependence. PMID- 11436130 TI - Genetic association studies of schizophrenia using the 8p21-22 genes: prepronociceptin (PNOC), neuronal nicotinic cholinergic receptor alpha polypeptide 2 (CHRNA2) and arylamine N-acetyltransferase 1 (NAT1). AB - Schizophrenia is a common, genetically heterogeneous disorder with a lifetime prevalence of approximately 1% in the general population. Linkage studies of affected families have now strongly implicated a susceptibility locus on chromosome 8p21-22. Tests of allelic association with markers on 8p21-22 should be able to localise any quantitative trait nucleotides (QTN's) or susceptibility mutations to within a few hundred kilobases. Three brain expressed candidate susceptibility genes, prepronociceptin (PNOC), neuronal cholinergic receptor, nicotinic, alpha polypeptide 2 (CHRNA2) and arylamine N-acetyltransferase 1 (NAT1) have been mapped to chromosome 8p21-22. A case-control, allelic association study was performed using a novel highly polymorphic dinucleotide repeat, D8S2611 near the PNOC gene, two previously characterised dinucleotide repeats, D8S131 and D8S131P at the CHRNA2 locus and an RFLP at the 3'UTR of the arylamine N-acetyltransferase 1 (NAT1) gene. No differences were found in allele frequencies between the patient and control groups. DNA variations or mutations at or near the three genes under study are unlikely to increase susceptibility to schizophrenia in our population sample. PMID- 11436131 TI - Modification of morphology and function of integument mitochondria-rich cells in tilapia larvae (Oreochromis mossambicus) acclimated to ambient chloride levels. AB - Similar to those of the gills of adults, three types of mitochondria-rich (MR) cells with different morphologies of apical surfaces (wavy convex, shallow basin, and deep hole) were identified on the integument of freshwater-acclimated tilapia larvae (Oreochromis mossambicus). The object of this study is to test the hypothesis that these subtype cells may represent MR cells equipped with variable efficiencies in Cl(-) uptake. Larvae acclimated to low-Cl(-) =0.001-0.007 mM) water developed higher densities of MR cells than those acclimated to high-Cl(-) =7.3-7.9 mM) water. The percentage of wavy-convex-type cells in total MR cells was higher in low-Cl(-)-acclimated larvae than in high-Cl(-)-acclimated larvae, which displayed only deep-hole type. In addition, Cl(-) influx rates of whole larva measured with (36)Cl(-) showed a coincident correlation with MR cell densities, that is, low-Cl(-) larvae displayed higher Cl(-) influx rates than did high-Cl(-) larva, suggesting that tilapia larvae develop a higher density of MR cells with larger apical surfaces (wavy-convex type) to boost Cl(-) uptake in Cl( )-deficient water. The distinct types of apical surfaces may represent different phases of MR cells that possess different efficiencies of Cl(-) uptake. Increased apical membrane surface areas of MR cells may provide larvae with rapid regulation of Cl(-) before new MR cells differentiate. PMID- 11436132 TI - Extracellular carbonic anhydrase in the dogfish, Squalus acanthias: a role in CO2 excretion. AB - In Pacific spiny dogfish (Squalus acanthias), plasma CO(2) reactions have access to plasma carbonic anhydrase (CA) and gill membrane-associated CA. The objectives of this study were to characterise the gill membrane-bound CA and investigate whether extracellular CA contributes significantly to CO(2) excretion in dogfish. A subcellular fraction containing membrane-associated CA activity was isolated from dogfish gills and incubated with phosphatidylinositol-specific phospholipase C. This treatment caused significant release of CA activity from its membrane association, a result consistent with identification of the dogfish gill membrane bound CA as a type IV isozyme. Inhibition constants (K(i)) against acetazolamide and benzolamide were 4.2 and 3.5 nmol L(-1), respectively. Use of a low dose (1.3 mg kg(-1) or 13 micromol L(-1)) of benzolamide to selectively inhibit extracellular CA in vivo caused a significant 30%-60% reduction in the arterial venous total CO(2) concentration difference, a significant increase in Pco(2) and an acidosis, without affecting blood flow or ventilation. No effect of benzolamide on any measure of CO(2) excretion was detected in rainbow trout (Oncorhynchus mykiss). These results indicate that extracellular CA contributes substantially to CO(2) excretion in the dogfish, an elasmobranch, and confirm that CA is not available to plasma CO(2) reactions in rainbow trout, a teleost. PMID- 11436133 TI - The ontogeny of pulmonary surfactant secretion in the embryonic green sea turtle (Chelonia mydas). AB - Pulmonary surfactant, consisting predominantly of phosphatidylcholine (PC), is secreted from Type II cells into the lungs of all air-breathing vertebrates, where it functions to reduce surface tension. In mammals, glucocorticoids and thyroid hormones contribute to the maturation of the surfactant system. It is possible that phylogeny, lung structure, and the environment may influence the development of the surfactant system. Here, we investigate the ontogeny of PC secretion from cocultured Type II cells and fibroblasts in the sea turtle, Chelonia mydas, following 58, 62, and 73 d of incubation and after hatching. The influence of glucocorticoids and thyroid hormones on PC secretion was also examined. Basal PC secretion was lowest at day 58 (3%) and reached a maximal secretion rate of 10% posthatch. Dexamethasone (Dex) alone stimulated PC secretion only at day 58. Triiodothyronine (T(3)) stimulated PC secretion in cells isolated from days 58 and 73 embryos and from hatchling turtles. A combination of Dex and T(3) stimulated PC secretion at all time points. PMID- 11436134 TI - Effect of air breathing on acid-base and ion regulation after exhaustive exercise and during low pH exposure in the bowfin, Amia calva. AB - To explore a potential conflict between air breathing and acid-base regulation in the bowfin (Amia calva), we examined how individuals with access to air differed from fish without air access in their response to acidosis. After exhaustive exercise, bowfin with access to air recovered significantly more slowly from the acidosis than fish without air access. While arterial blood pH (pH(a)) of fish without air access recovered to resting levels by 8 h, pH(a) was still significantly depressed in fish having access to air. In addition, Pco(2) was slightly more elevated in fish having air access than those without it. Fish with access to air still had a significant metabolic acid load after 8-h recovery, while those without air access completely cleared the load within 4 h. These results suggest that bowfin with access to air were breathing air and, consequently, were less able to excrete CO(2) and H(+) and experienced a delayed recovery. In contrast, during exposure to low pH, air breathing seemed to have a protective effect on acid-base status in bowfin. During exposure to low pH water, bowfin with access to air developed a much milder acidosis than bowfin without air access. The more severe acidosis in fish without air access was caused by an increased rate of lactic acid production. It appears that enhanced O(2) delivery allowed air-breathing bowfin to avoid acidosis-induced anaerobic metabolism and lactic acid production. In addition, during low pH exposure, plasma Na(+) and Cl( ) concentrations of fish without air access fell slightly more rapidly than those in fish with air access, indicating that the branchial ventilatory changes associated with air breathing limited, to some degree, ion losses associated with low pH exposure. PMID- 11436135 TI - Cold-hardiness and evaporative water loss in hatchling turtles. AB - North American turtles hatch in late summer and spend their first winter either on land or underwater. Adaptations for terrestrial overwintering of hatchlings in northern regions, where winter thermal and hydric regimes are harsh, have not been systematically investigated in many species. We measured intrinsic supercooling capacity, resistance to inoculative freezing, and desiccation resistance in hatchlings of terrestrial and aquatic turtles collected from northern (Terrapene ornata, Chrysemys picta bellii, Kinosternon flavescens, Chelydra serpentina) and southern (Chrysemys picta dorsalis, Trachemys scripta, Sternotherus odoratus, Sternotherus carinatus) locales. Supercooling capacity was estimated from the crystallization temperature of turtles cooled in the absence of external ice nuclei. Mean values ranged from -8.1 degrees to -15.5 degrees C and tended to be lower in terrestrial hibernators. Inoculation resistance was estimated from the crystallization temperature of turtles cooled in a matrix of frozen soil. These values (range of means: -0.8 degrees to -13.6 degrees C) also tended to be lower in the terrestrial hibernators, especially C. picta bellii. Mean rates of evaporative water loss varied markedly among the species (0.9-11.4 mg g(-1) d(-1)) and were lowest in the terrestrial hibernators. Most species tolerated the loss of a modest amount of body water, although half of the sample of S. carinatus died from desiccation. In general, turtles did not regain lost body water from wet soil, and immersion in free water was required for rehydration. Therefore, desiccation resistance may be an important adaptation to terrestrial hibernation. Resistances to inoculative freezing and desiccation were directly correlated, perhaps because they are governed by the same morphological characteristics. PMID- 11436136 TI - Effects of age on water balance in Drosophila species. AB - Age-related declines in physiological performance have been documented in a wide variety of organisms. However, it is unknown whether related species age in a similar manner or whether physiological differences associated with aging differ widely among species. Previous work has shown that the desiccation resistance of Drosophila melanogaster decreases rapidly with age. Other Drosophila species may have delayed reproductive maturity or may inhabit arid habitats, so that age related changes in water balance may be a significant physiological constraint on their ecological success. We may, therefore, predict that physiological systems involved in water balance will deteriorate with age at a reduced rate in these species. We quantified several components of water budgets for 14 species of Drosophila, including both desert and mesic representatives differing in their age of maturity. Desiccation resistance decreased with age in all but one species studied, primarily because of increased rates of water loss. However, there was no significant relationship between the rate of aging, as indicated by the rate of increase in water-loss rates as flies aged, and either habitat or age of maturity. PMID- 11436137 TI - Postweaning duration and body composition changes in Southern elephant seal (Mirounga leonina) pups at King George Island. AB - Weaning mass in southern elephant seals is highly variable, the heaviest pups being three times as heavy as the lightest ones. After weaning, pups undergo an extensive postweaning period in which they draw on their reserves. To quantify the energy expenditure during the postweaning period, changes in mass, body composition, and postweaning duration were measured in southern elephant seals at King George Island, South Shetland Islands, Antarctica. Overall, mean pup weaning mass was 154 +/- 26 kg (n=117) and did not differ between sexes. Mean minimum postweaning duration was 42.5 +/- 7.5 d. Heavier animals at weaning had lower mass-specific mass loss rates than lighter ones, and a faster depletion of body reserves was associated with a shorter postweaning period. The proportion of body mass represented by fat at weaning was 37% +/- 4% (n=47) and did not differ between sexes. Of these pups, 36 were recaptured after a mean period of 36 d after weaning. On average, total mass loss measured in these animals (39 kg) was composed of 39% water, 47% fat, and 12% protein. The composition of mass loss was not significantly different between sexes and was not related to weaning mass or total body energy reserves. However, fatter animals at weaning lost more fat per kilogram lost than thinner ones. Late in the fast, males and females appeared to be in a similar body condition. Nevertheless, the overall proportion of body mass represented by fat at this time was lower than that presented by the same animals at weaning. We estimated that during the postweaning period pups lost, on average, 30% of their mass at weaning. This comprised approximately 35% of the energy and 32% of the fat in the pup's body. PMID- 11436138 TI - Energetic cost of foraging in free-diving emperor penguins. AB - Hypothesizing that emperor penguins (Aptenodytes forsteri) would have higher daily energy expenditures when foraging for their food than when being hand-fed and that the increased expenditure could represent their foraging cost, we measured field metabolic rates (FMR; using doubly labeled water) over 4-d periods when 10 penguins either foraged under sea ice or were not allowed to dive but were fed fish by hand. Surprisingly, penguins did not have higher rates of energy expenditure when they dove and captured their own food than when they did not forage but were given food. Analysis of time-activity and energy budgets indicated that FMR was about 1.7 x BMR (basal metabolic rate) during the 12 h d( 1) that penguins were lying on sea ice. During the remaining 12 h d(-1), which we termed their "foraging period" of the day, the birds were alert and active (standing, preening, walking, and either free diving or being hand-fed), and their FMR was about 4.1 x BMR. This is the lowest cost of foraging estimated to date among the eight penguin species studied. The calculated aerobic diving limit (ADL(C)), determined with the foraging period metabolic rate of 4.1 x BMR and known O(2) stores, was only 2.6 min, which is far less than the 6-min ADL previously measured with postdive lactate analyses in emperors diving under similar conditions. This indicates that calculating ADL(C) from an at-sea or foraging-period metabolic rate in penguins is not appropriate. The relatively low foraging cost for emperor penguins contributes to their relatively low total daily FMR (2.9 x BMR). The allometric relationship for FMR in eight penguin species, including the smallest and largest living representatives, is kJ d( 1)=1,185 kg(0.705). PMID- 11436139 TI - Milking strategy in subantarctic fur seals Arctocephalus tropicalis breeding on Amsterdam Island: evidence from changes in milk composition. AB - Milk composition was investigated throughout the 10-mo pup-rearing period in subantarctic fur seals (Arctocephalus tropicalis) breeding on Amsterdam Island. The mean milk composition was 42.8% +/- 5.7% lipid, 12.1% +/- 1.5% protein, and 42.6% +/- 7.3% water. Subantarctic fur seals breeding on Amsterdam Island produced one of the richest milks ever reported in otariids (20.4 +/- 2.9 kJ/g), with lipid content contributing 85% of total gross energy. The high lipid levels measured in the milk of subantarctic fur seals breeding on Amsterdam Island is consistent (i) with the relatively long time lactating females spend at sea, due to the relatively poor local trophic conditions near the colony that necessitate that they travel long distances to reach the foraging grounds, and (ii) with the consequently short time mothers spend with their pups ashore. Milk composition changed according to the time mothers were fasting ashore: milk produced during the first 2 d spent ashore, when more than 80% of milk transfer occurred, had higher levels of lipids, proteins, and gross energy than milk produced later during the visit ashore, suggesting that the pups were fed with two types of milk during a suckling period. Throughout the year, mothers in good condition produced milk of higher lipid content than others, suggesting that individual foraging skills contribute to enhance milk quality. Milk lipid and gross energy content varied with pup age, according to quadratic relationships, increasing during the earlier stages of lactation before reaching asymptotic values when pups were 180 d old. The stage of lactation appears to be a better predictor of milk lipid content than the duration of the preceding foraging trip, suggesting that either changes in the nutritional requirements of the pup and/or seasonal changes in trophic conditions act on milk composition. These changes in milk quality may also be related to changes in maternal care; lactating subantarctic fur seals apparently reallocate their body reserves toward gestation rather than lactation at the end of the pup-rearing period. PMID- 11436140 TI - Nutrient uptake by embryos of the Australian viviparous lizard Eulamprus tympanum. AB - Eulamprus tympanum is a high-altitude viviparous lizard that was probably used to help define a Type I chorioallantoic placenta. In this article, we (1) describe the net transport of nutrients across the placenta of E. tympanum, and (2) compare placental uptake in E. tympanum with a previous study of Eulamprus quoyii, which occurs in warmer environments, to assess the potential importance of thermal regime on placentotrophy. Freshly ovulated eggs are 387.3+/-19.7 mg. There is a significant net uptake of water and a net loss of dry matter during development, so the dry neonate is only 84% the size of the dry egg. There is no significant change in the total ash or nitrogen in eggs during embryonic development, with the entire loss of dry matter being lipid. Almost the entire loss of lipid occurs in the triacylglycerol fraction, with no net change in phospholipids. A net increase in total cholesterol suggests that cholesterol is synthesised by the developing embryo. The lipid profile of eggs of E. tympanum reflects that of other species with simple placentae in having a relatively high proportion of triacylglycerol and little cholesterol. The fatty acid composition of eggs reflects that expected in the diet of E. tympanum. There is a preservation and some synthesis of arachidonic (20:4n-6) and docosahexaenoic (22:6n-3) acids in the phospholipid fraction during embryonic development. Despite there being no net uptake of ash, there is a net increase in calcium, potassium, sodium, and magnesium in the neonate compared with the egg. We conclude that E. tympanum, like E. quoyii, is predominantly lecithotrophic with little, if any, uptake of organic molecules but with significant uptake of some inorganic ions and water. In addition, there is no difference in placentotrophy correlated with differences in the environments inhabited by each species. PMID- 11436141 TI - Seasonal acclimatization to extreme climatic conditions by black-capped chickadees (Poecile atricapilla) in interior Alaska (64 degrees N). AB - Winters in interior Alaska (64 degrees N) are characterized by short photoperiod (5L : 19D) and chronic subfreezing temperatures. To determine if seasonal acclimatization of black-capped chickadees (Poecile atricapilla) at high latitude differs from that of conspecifics at lower latitudes, standard metabolic rates (SMR), metabolic response to low temperature (-30 degrees C), nocturnal hypothermia, body mass, fat reserves, and conductance were measured over two winters and one summer in three groups of seasonally acclimatized birds. Body mass and conductance did not vary with season, although furcular fat levels were higher in winter. Birds used nocturnal hypothermia when exposed to -30 degrees C in summer or winter. Although SMR did not vary seasonally, winter SMRs differed between the two winters of the study. Nocturnal hypothermia in summer and decreased SMR in response to extreme conditions may either reflect plasticity inherent to all populations of black-capped chickadees or may result from individual variation within this northern population. PMID- 11436142 TI - Angiotensin II alters blood flow distribution in amphibians. AB - In toads, angiotensin II (ANG II) induces the water absorption response (WR) during which the seat patch (pelvic+inner-thigh skin) is pressed to a wet substrate from which water flows osmotically into the animal. Since ANG II is a potent vasoconstrictor, it has the potential to redistribute blood flow. To determine the regional circulatory effects of ANG II, we used microsphere methods to measure relative changes in blood flow to several skin regions and other organs before and after ANG II administration in terrestrial toads and aquatic bullfrogs. In toads, after ANG II administration, seat patch and bladder blood flow increased by 264.2%+/-197.6% and 287.2%+/-86.7%, respectively (P<0.05), while dorsal and pectoral skin flow decreased by 48.0%+/-19.4% and 21.3%+/-25.4%, respectively (P<0.05). In bullfrogs, ANG II caused no significant changes in blood flow. Our results support our hypothesis that, in toads, ANG II increases and decreases blood flow to regions of the body associated with water gain and water loss, respectively. PMID- 11436143 TI - The membrane-bound intestinal enzymes of waxwings and thrushes: adaptive and functional implications of patterns of enzyme activity. AB - Cedar waxwings (Bombycilla cedrorum) feed predominantly on fruits that are rich in simple sugars and low in nitrogen, supplementing this diet with arthropod prey during the summer months as well as flowers and tree sap in springtime. In contrast, thrushes feed extensively on fatty, protein-rich invertebrate prey, supplemented with sugary and lipid-rich fruits. Simple sugars and fats are digested and/or absorbed by distinctly different physiological mechanisms, which suggests the possibility of contrasting digestive strategies in animals specialized to diets containing one of these two energy sources. In this study, we quantified enzymatic activity of three membrane-bound intestinal enzymes of cedar waxwings and five species of thrushes to explore this aspect of their digestive physiology. These enzymes catalyze the final steps in the digestion of carbohydrates (sucrase-isomaltase and maltase-glucoamylase) and protein (aminopeptidase-N). The two carbohydrases are homologous enzymes with overlapping functions; both enzymes catalyze the hydrolysis of maltase and isomaltase. The membrane-bound digestive enzyme systems that we described for cedar waxwings and thrushes can be explained by the particular nutrients contained within their respective natural diets. Consistent with previous work, cedar waxwings displayed intestinal sucrase activity, whereas thrushes did not. Correspondingly, cedar waxwings eat some foods containing sucrose, whereas thrushes do not. Sucrase isomaltase conferred all maltase and isomaltase activity in cedar waxwings. In contrast, all maltase and isomaltase activity in thrushes was necessarily sucrase independent, which indicated the presence of maltase-glucoamylase. The absence of sucrase-independent maltase activity in cedar waxwings suggests that sucrase isomaltase obviates the need for maltase-glucoamylase. Indeed, total maltase and isomaltase activities were much higher in cedar waxwings than in thrushes. Neither waxwings nor thrushes eat starchy foods; sucrase-isomaltase in waxwings and maltaseglucoamylase in thrushes probably function in digesting glycogen in animal foods. We suggest that digestive traits associated with specialization to monosaccharide-rich fruits (lack of a grinding gizzard) by frugivorous waxwings and thrushes may prevent utilization of starchy seeds. Total aminopeptidase-N activity in cedar waxwings was indistinguishable from the allometric pattern among thrush species, but the distribution of this enzyme along the intestines of waxwings and thrushes was distinctly different, which demonstrates that total enzyme activity can be insufficient as a descriptor of the functional activity of brush border enzymes. Aminopeptidase-N activity peaked in the anterior part of the intestines of thrushes and in the terminal portion of the intestines of waxwings, which suggests contrasting strategies for protein digestion from fatty versus sugary diets, respectively. PMID- 11436144 TI - Contrasting responses to desiccation and starvation by eggs and neonates of two lepidoptera. AB - We examined the effects of desiccation on eggs and first-instar larvae of two species of Lepidoptera, Grammia geneura (Arctiidae) and Manduca sexta (Sphingidae). Grammia geneura occurs primarily in grasslands and savannas of the southwestern United States; M. sexta co-occurs with G. geneura but also is cosmopolitan across much of the Western Hemisphere. Eggs of G. geneura exposed to 0% relative humidity (RH) lost water much less rapidly (7.6 microg d(-1); 2.4% d( 1)) than did eggs of M. sexta (79.5 microg d(-1); 5.7% d(-1)). Eggs of both species survived at rates exceeding 75% at both 0% and 85% RH. Neonates of the two species responded differently to desiccation and starvation. In 85% RH, larval G. geneura survived at high rates (>80%) without access to food or water up to day 17, and in 0% RH, they survived at rates exceeding 50% through the first 10 d. Larvae at 0% RH lost mass very slowly (7.2 microg d(-1); 2.9% d(-1)), which was attributable both to low rates of water loss and to an ability to reduce metabolic rate to low levels. Larval M. sexta, in contrast, had rates of mortality that were much higher: after 1 d, fewer than 30% were alive in either group, and by about 1.5 d, all were dead. Neonate M. sexta also lost mass much more rapidly at 0% RH, about 329 microg d(-1). Water from metabolism appeared to contribute significantly to the water budget of G. geneura but not of M. sexta. These data show that G. geneura and M. sexta can inhabit similar macroclimates via remarkably different physiologies. PMID- 11436145 TI - Differences in pH between interstitial fluid and arterial blood in water breathing and air-breathing vertebrates. AB - Most cells are bathed by interstitial fluid, but extracellular pH measurements are mostly for arterial plasma. Whole-body mean pH differences between the two fluids have been estimated in terms of a simple model. This relates to the diffusive exchange of carbon dioxide and oxygen and utilizes literature data, for 22 vertebrate species, on arterial and mixed-venous tensions of both gases. Uncertainties arise because the carbon dioxide reaction in blood may sometimes be in disequilibrium and because carbon dioxide diffusion is facilitated to unknown degrees in the presence of buffers. Nevertheless, the model suggests that the pH difference should tend to vary inversely with arterial carbon dioxide tension. In some species, this may aid interstitial pH homeostasis, but a clearer implication is that the difference should be generally greater in water breathers than in air breathers. It has previously been found that arterial pH in water-breathing teleosts also tends to be higher than in air-breathing tetrapods (when allowance is made for temperature and plasma sodium concentration) and to a comparable extent. Thus, mean interstitial pH may be more nearly similar in the two groups than is arterial pH. Direct measurements of interstitial pH do not yet suffice to test the model. PMID- 11436146 TI - On the presentation of statistical tests of place: the importance of editorial consistency. PMID- 11436147 TI - Epithelial membrane antigen and S-100 protein expression in benign and malignant papillary thyroid neoplasms. AB - Papillary carcinoma of the thyroid is mainly diagnosed with histopathologic features. Classical papillary architectures are important but nuclear change is the essential diagnostic element. Papillary architecture may be seen in benign lesions such as in hyperplastic areas of the follicular neoplasms, multinodular goiter and Grave's disease. Differential diagnosis of papillary carcinoma and papillary hyperplasia is very important for clinical management. Some authors have reported that Epithelial Membrane Antigen (EMA) and S-100 protein expression would be valuable and helpful in identifying papillary neoplasia and distinguishing it from papillary hyperplasia. In this study, EMA and S-100 protein expression of 14 papillary thyroid carcinomas and 13 papillary hyperplasias were studied by using immunohistochemical methods. In 14 papillary carcinomas, 9 showed diffuse and 3 revealed focal S-100 protein nuclear and cytoplasmic immunostaining. Two cases were not stained. All of the 13 papillary hyperplasias were negative for S-100 protein. EMA expression was observed in the apical cytoplasmic location of 11 papillary carcinomas except one case that showed diffuse cytoplasmic staining and one which was negative. In the papillary hyperplasias, 7 revealed both cytoplasmic and apical cytoplasmic staining. One case showed only cytoplasmic staining. Five cases were negative for EMA. The difference in the S-100 protein expression is significant, however immunostaining of EMA is similar in both lesions. We concluded that differential diagnosis of papillary structures in carcinomas and hyperplasias was mainly diagnosed on the histopathologic features but S-100 protein expression could be helpful in difficult cases. PMID- 11436148 TI - Congenital epulis: an ultrastructural and immunohistochemical case study. AB - We report a case of an otherwise healthy, two days old, female with a a mass derived from the mandible. Light microscopy findings were compatible with the diagnosis of congenital epulis. The lesion showed negative staining for S-100 protein, which is a characteristic finding in cases of epulis and distinguishes it from granular cell tumor. The characteristic electron microscopy findings, as well as other features of epulis in relation to the theories for its possible origin are also reviewed PMID- 11436149 TI - Sirenomelia with esophageal atresia. AB - Sirenomelia, is a rare fatal condition characterized by fusion of the lower extremities. It has been suggested that sirenomelia is a severe form of caudal regression syndrome complex for which the pathogenesis is controversial. Our case is a sirenomelia associated with esophageal atresia and tracheoesophageal fistula. In the literature, it has been pointed out that the VATER association may represent a less severe form of sirenomelia but no sirenomelia case associated with esophageal atresia and tracheoesophageal fistula has been found. Finally, we detected a single large artery which diverts the blood of the embryo's caudal part to the placenta with the detailed autopsy especially with focusing on the abdominal vasculature. This vascular steal is thought to be the main pathogenic mechanism of the condition. PMID- 11436150 TI - Adenocarcinoma of the rete testis. Report of two cases. AB - Adenocarcinoma of the rete testis is extremely rare and has been the subject of only sporadic case reports. Its natural history appears highly malignant even in localized forms; because of a poor response to adjuvant therapy radical orchiectomy with retroperitoneal lymph node dissection is recommended in stage A. We report our experience with two examples which arose in men of 26 and 57 years of age between 1988 and 1999. Both of them presented with a scrotal mass, and one of them had hydrocele. Neither had a history of maldescendent, infections or trauma. Microscopically the tumors were adenocarcinomas, and the one in the 26 years old patient presented with tubulopapillary formations. Immunohistochemical staining demonstrated positivity for CK and NSE in the 26 year old patient and only a weak positivity for CEA and vimentin in the 57 year old one. EMA and calretinin were always negative. Both were stage A but the young man who underwent radical orchiectomy with no lymph node dissection, developed a lymph node metastasis within one year. PMID- 11436151 TI - Pleural malignant mesothelioma. PMID- 11436152 TI - Mesothelioma: diagnostic immunohistochemical markers. PMID- 11436153 TI - The role of immunohistochemistry in the differential diagnosis of serous effusions. PMID- 11436154 TI - Prognostic markers in malignant mesothelioma. PMID- 11436155 TI - The plasmacytoid monocyte: from morphology to function. PMID- 11436156 TI - Marginal zone lymphomas, MALT and splenic types. PMID- 11436157 TI - A histogenetic model for Hodgkin's lymphoma development. A Proposal based on immunophenotypic markers. PMID- 11436158 TI - Molecular interactions in copper metabolism. PMID- 11436159 TI - Cytokeratin immunohistochemistry in liver histopatology. PMID- 11436160 TI - Inborn errors of metabolism and hepatocellular carcinoma (HCC). PMID- 11436161 TI - Epithelial tumors of the adult kidney: evaluation of the current diagnostic and prognostic criteria. PMID- 11436162 TI - Differential diagnosis of a testicular mass in an adult. PMID- 11436163 TI - p27KIP1: androgen regulation and prognostic significance in prostate cancer. PMID- 11436164 TI - Applications of comparative genomic hybridization in surgical pathology. PMID- 11436165 TI - Confocal laser scanning microscopy. PMID- 11436166 TI - A clinicopathologic and immunohistochemical study of 35 anaplastic carcinomas of the pancreas with a review of the literature. AB - Anaplastic pancreatic carcinomas are rare tumors, frequently displaying a variety of growth patterns. The literature lacks a comprehensive study of this tumor. Thirty-five cases of anaplastic carcinoma of the pancreas diagnosed between 1955 and 1997 were retrieved from the Endocrine Registry at the Armed Forces Institute of Pathology. Histology, immunophenotype, molecular analysis, and patient follow up were analyzed. The tumors of 10 women and 25 men, aged 34 to 85 years (mean age at presentation, 62.5 years), were studied. Patients had vague symptoms (weight loss, pain, and fatigue, nausea, or vomiting), lasting an average of 13.2 weeks. The tumors, of an average size of 9.2 cm, were usually in the head or tail of the pancreas. The tumors were widely infiltrative, histomorphologically separated into predominantly large, pleomorphic cell, or spindle cell groups. Tumor phagocytosis and necrosis were noted. Immunohistochemical studies confirmed an epithelial origin with at least one epithelial marker in 78% of the tumors. K ras mutations by sequence analysis were found in eight of 12 cases tested. Surgical biopsy/excision was used in all patients. Twenty-nine of 35 patients died of disease (average, 5.2 months), three died with no evidence of disease (average, 56.9 months), and three patients were alive at last follow-up (average, 94.0 months), one with residual disease. There was no statistically significant difference in survival between patients with and without a K-ras mutation. Anaplastic carcinoma of the pancreas usually occurs in the head of the pancreas in older men. The epithelial nature of the pleomorphic cells (giant or spindled) can usually be documented. Patients with K-ras mutations have a shorter survival time, even though the overall prognosis for all anaplastic carcinomas is fatal (93% fatality; average survival, 448 days). Ann Diagn Pathol 5: 129-140, 2001. This is a US government work. There are no restriction on its use. PMID- 11436167 TI - Intraglandular necrotic debris in gastric biopsy and surgical specimens. AB - To clarify whether intraglandular necrotic debris (IND) is a diagnostic clue to noninvasive high-grade neoplasia or invasive neoplasia of the stomach, we reviewed 135 gastric biopsy specimens and 55 surgical materials. Intraglandular necrotic debris is defined as an eosinophilic material with necrotic epithelial fragments within the lumen of a dilated atypical gland. Using the Vienna classification, the incidence of IND in category 4 (noninvasive high-grade neoplasia) and category 5 (invasive neoplasia) was significantly higher than that of category 1 (negative for neoplasia/dysplasia), category 2 (indefinite for neoplasia/dysplasia), and category 3 (noninvasive low-grade neoplasia). The incidence of IND was much higher in category 5 than in category 4 in biopsy specimens. In addition, cases with IND in category 4 in biopsy specimens turned out to be either carcinoma in situ or invasive carcinoma in the surgical specimens. According to the histologic classification of surgically removed invasive carcinoma, moderately differentiated adenocarcinoma showed the highest incidence of IND. Intraglandular necrotic debris was not found in either signet ring cell carcinoma or mucinous adenocarcinoma. Our results indicate that IND in biopsy specimens is a diagnostic clue to noninvasive high-grade neoplasia or invasive carcinoma, and the origin of IND may be associated with necrotic atypical epithelium. PMID- 11436168 TI - Clinicopathologic study of retinoblastoma including MIB-1, p53, and CD99 immunohistochemistry. AB - Retinoblastoma is the most common intraocular tumor of childhood and has served as a model for the understanding or tumorigenesis. This study retrospectively examines the clinicopathologic features of 19 retinoblastomas and defines the MIB 1 (cell proliferation marker), p53 (tumor suppression gene), and CD99 (HBA71 or MIC2 antibody) immunoreactivity in 10 selected cases. Nineteen patients (11 boys), ranging in age from 6 to 47 months (mean, 20 months), were included for study. Clinical presentations included: leukocoria (n = 12), strabismus (n = 6), apparent decreased visual acuity (n = 5), and proptosis (n = 1). Five patients had bilateral tumors and one neoplasm arose in a patient with a known family history of retinoblastoma. All tumors were histologically characterized by a proliferation of small cells with high nuclear-to-cytoplasmic ratios. Commonly encountered histologic features included necrosis (n = 17, 89%), calcification (n = 16, 84%), fleurettes (n = 14, 74%), and Flexner-Wintersteiner rosettes (n = 11, 58%). Retinal involvement was noted in 18 tumors (95%) and optic nerve invasion in six cases (32%). The surgical optic nerve margin was positive in one case. Mitosis counts were evaluable in 18 cases and ranged from 1 to 42 mitotic figures/10 high power field (mean, 13 mitotic figures/10 high power field). Ten tumors were evaluated with MIB-1, p53, and CD99 antibodies by paraffin immunohistochemistry. MIB-1 labeling indices ranged from 31.4 to 77.1 (mean, 49.4). p53 immunostaining was observed in six tumors; less than 10% of tumor cells were noted to be p53 positive in each case. CD99 positivity was demonstrable focally in three tumors. Adjuvant chemotherapy and/or radiation therapy was administered in six patients. Tumor recurrence was not observed in any of the patients with a mean follow-up of 8.9 years. Only one patient died (20 years after enucleation) because of metastatic osteosarcoma. IN CONCLUSION: (1) Fleurettes and Flexner-Wintersteiner rosettes are variable findings in retinoblastoma. (2) Retinoblastomas are characterized by marked cell proliferation as evidenced by generally high mitosis counts and extremely high MIB-1 labeling indices, but this does not appear to adversely impact on prognosis. (3) Unlike peripheral primitive neuroectodermal tumors, most retinoblastomas do not stain positively with antibody to CD99. (4) Limited p53 immunostaining was present in 60% of tumors studied. (5) Enucleation with negative optic nerve margin is potentially curative in patients with retinoblastoma. PMID- 11436169 TI - Malignant mixed tumor ex eccrine spiradenoma: an unusual pattern of malignant dedifferentiation. AB - Eccrine spiradenoma (ES) is a benign tumor of the skin adnexal origin. It is often seen in the head and neck region of young adults and may be present for years. While there have been numerous case reports of malignant degeneration within ES, they have been mostly carcinomatous dedifferentiation and rarely carcinosarcomas. A malignant mixed tumor is a neoplasm with a malignant epithelial component and areas of chondroid and myxoid differentiation within the malignant epithelial proliferation. While the epithelial component is malignant, the mesenchymal component is felt to represent a benign, metaplastic response of the stroma. While the malignant mixed tumor has a benign counterpart, the benign mixed tumor, the former is usually not seen in continuity with the latter. We describe a case of ES with malignant degeneration and demonstrate the transformation from benign to malignant. The histology and immunohistochemistry of the neoplasm supports a malignant mixed tumor, an extremely unusual neoplasm to see in association with ES or any benign adnexal neoplasm. PMID- 11436170 TI - Ki-1 anaplastic large-cell lymphoma occurring at the site of ileocolonic anastomosis in a patient treated surgically for colonic adenocarcinoma: case report and review of the literature. AB - Systemic anaplastic large-cell lymphoma is an uncommon type of non-Hodgkin lymphoma characterized by strong expression of the Ki-1 (CD30) antigen. Gastrointestinal involvement typically is less common than in other types of non Hodgkin's lymphoma. We report a case of CD30-positive anaplastic large-cell lymphoma occurring at the site of colonic anastomosis in an elderly patient who had been treated for colonic adenocarcinoma by right hemicolectomy 10 years previously. The lymphoma was a 2-cm mass composed of large, atypical cells infiltrating the mucosa, submucosa, and muscularis propria. Immunoperoxidase stain was strongly positive for Ki-1, and negative for LeuM1, L26, UCHL1, EMA, and cytokeratin. There have been numerous reports of unusual extranodal presentations of systemic anaplastic large-cell lymphoma; the only previously reported case involving the colon, however, occurred in the context of ulcerative colitis. Anastomotic recurrence is a relatively common complication of surgical therapy for adenocarcinoma, but the recurrent tumors are invariably adenocarcinomas. We are aware of no cases of lymphoma of any type occurring at the site of anastomosis after resection for adenocarcinoma. PMID- 11436171 TI - Deciduoid mesothelioma: a rare, distinct entity with unusual features. AB - An unusual case of deciduoid mesothelioma occurring in the anterior abdominal wall of a 30-year-old woman is reported. The patient had a palpable mass that was resected. The mass appeared largely cystic with solid areas. Histologically, the tumor cells appeared epitheloid with eosinophilic cytoplasm and prominent nucleoli. The tumor was positive for keratins and vimentin and negative for CEA and Ber-EP4. Electron microscopy showed features of mesothelial cells characterized by well-formed desmosomes and long, slender microvilli. In contrast to previously reported cases of deciduoid mesothelioma, this tumor developed in the abdominal wall and appears to have a benign course. PMID- 11436172 TI - Dermoid cyst of the maxillary sinus. AB - Dermoid cysts are rare developmental teratomatous lesions composed of ectodermally derived stratified squamous epithelium and mesodermally derived skin adnexal structures. As part of the teratomatous lesion group, dermoid cysts are related to teratoid cysts, true teratomas, and epignathi. Although several theories have been postulated, the pathogenesis of dermoid cysts, and teratomatous lesions in general, is unclear. Most commonly affecting sites within the head and neck, dermoid cysts may be found in the frontotemporal/lateral brow area, central nasal area, oral cavity, lateral neck, and other sites. We present what is believed to be the first reported case arising within the maxillary sinus and briefly discuss the possible pathogenesis. PMID- 11436173 TI - When to suspect hypercoagulability and how to investigate it. AB - The authors summarize the current knowledge in the area of novel risk factors for venous thrombosis, both genetic and acquired causes of hypercoagulability. Because the list of genetic defects that predispose carriers to develop thrombosis has increased significantly in recent years, along with the available assays to test for them, hypercoagulability has been a subject of much discussion. This review highlights the issues on hypercoagulability that pathologists who are not specialists in coagulation should be familiar with, in order to provide consultation to other physicians. An educated approach to the diagnosis of this newly described group of disorders will ensure more cost effective and efficient care to patients at risk. PMID- 11436174 TI - The microbe, creator of the pathologist: an inter-related history of pathology, microbiology, and infectious disease. AB - This brief historical review of pathology stresses the impact of microbial discovery on the development of pathology as a medical specialty. If, as it has been said, the microscope invented the pathologist, it was the microbe, especially the pathogenic bacterium, that gave him his name and made him clinically relevant. PMID- 11436175 TI - Preliminary report: Amino acid profile in platelets of diabetic patients. AB - Platelet levels of 19 amino acids were measured in 20 outpatients with type 1 (age [mean +/- SE], 35.5 +/- 2.0 years) and 27 with type 2 (age, 58.4 +/- 1.4 years) diabetes, and 20 young (age 33.7 +/- 1.3 years) and 20 older (age 57.4 +/- 1.5 years) healthy volunteers. Platelet levels of most amino acids tended to be lower in patients with type 1 diabetes than in healthy controls. In particular, asparagine, glycine, taurine, alanine, valine, cysteine, leucine, phenylalanine, and lysine levels, expressed as nmol/10(8) platelets, were significantly lower. Only taurine significantly decreased in patients with type 2 diabetes, whereas threonine, alanine, and isoleucine increased. PMID- 11436176 TI - Review article: Regulation of glucose production with special attention to nonclassical regulatory mechanisms: a review. AB - Hepatic glycogenolysis and gluconeogenesis are essential processes for the prevention of hypoglycemia during short-term starvation. As has been calculated from stable isotope studies, gluconeogenesis accounts for approximately 35% to 50% of total basal glucose production, glycogenolysis for the other 50% to 65%. In long-term starvation, the kidney also contributes to glucose production by gluconeogenesis. Glucose production is regulated by the interaction of different regulatory mechanisms, eg, by glucoregulatory hormones, glucose itself, and gluconeogenic substrates. In the last decades, more insight has been gained into the importance of the autonomous nervous system and the existence of an extensive paracrine network in the liver that seems to exert a potent glucoregulatory role as well. This review is focused on the regulation of hepatic glucose production by the autonomous nervous system and the paracrine network, with special emphasis on studies carried out in human subjects. PMID- 11436177 TI - Moderate exercise, postprandial lipemia, and skeletal muscle lipoprotein lipase activity. AB - One mechanism by which prior exercise decreases the plasma triacylglycerol (TG) response to dietary fat may involve enhanced clearance of TG-rich lipoproteins. The purpose of the present study was to examine the influence of moderate intensity exercise on postprandial lipemia and muscle lipoprotein lipase (LPL) activity. Eight physically active, normolipidemic men aged 27.0 years (SD 4.2), body mass index 24.5 kg. m(-2) (SD 1.3), participated in 2 oral fat-tolerance tests with different preceding conditions. The afternoon before one test ( approximately 16 hours), subjects cycled for 90 minutes at 62.3% (SD 1.7%) of maximal oxygen uptake. Before the other test, subjects refrained from exercise. Samples of muscle, venous blood, and expired air were obtained in the fasted state. Subjects then consumed a high-fat meal (1.4 g fat, 1.2 g carbohydrate, 0.2 g protein, 73 kJ energy per kg body mass) before further blood and expired air samples were collected until 6 hours. The 6-hour areas under the TG concentration v time curves for plasma and for the chylomicron-rich fraction were lower (P <.05) after exercise (plasma, 7.91 [SE 1.09] v 5.72 [SE 0.47] mmol. L(-1). h; chylomicron-rich fraction, 1.98 [SE 0.51] v 0.92 [SE 0.16] mmol. L(-1). h). Muscle LPL activity was not significantly influenced by prior exercise, but the 4 subjects who had higher muscle LPL activity after exercise also had the most noticeable decreases in postprandial lipemia. The difference in lipemia between trials was inversely related to the difference in LPL activity (rho = -.79, P <.05). In the fasted state and postprandially, carbohydrate oxidation was lower after exercise (P <.05). Thus moderate exercise attenuates postprandial lipemia, possibly by altering muscle LPL activity. PMID- 11436178 TI - Effect of insulin, proinsulin, and amylin on renin release from perfused rat kidney. AB - To evaluate the possible role of insulin, proinsulin, and amylin in the renin angiotensin system, the direct effect of these peptides on renin release was examined using perfused kidney of rats. Renin release was significantly increased from a basal value of 6.1 +/- 1.8 to a peak value of 10.1 +/- 2.3 ng angiotensin I (Ang I)/mL/h by 0.5 nmol insulin, from 6.0 +/- 1.7 to 16.7 +/- 4.5 ng Ang I/mL/h by 1 nmol insulin, and from 6.1 +/- 1.8 to 18.0 +/- 5.5 ng Ang I/mL/h by 8 nmol insulin. Renin release was not significantly changed by perfusion of 0.05 nmol proinsulin or amylin but significantly increased from a basal value of 6.1 +/- 1.7 to a peak value of 8.1 +/- 3.6 ng Ang I/mL/h by 1 nmol proinsulin, from 5.6 +/- 1.7 to 12.1 +/- 3.8 ng Ang I/mL/h by 8 nmol proinsulin, from 5.7 +/- 1.9 to 8.2 +/- 3.5 ng Ang I/mL/h by 1 nmol amylin, and from 5.2 +/- 2.0 to 12.4 +/- 3.3 ng Ang I/mL/h by 8 nmol amylin. The concentration of cyclic adenosine monophosphate in the effluent was significantly increased from a basal value of 5.1 +/- 1.6 to a peak value of 10.6 +/- 2.5 mmol/min by 8 nmol amylin but not altered by perfusion of insulin or proinsulin. The addition of 0.05 nmol proinsulin and 0.05 nmol of amylin on 0.5 nmol insulin significantly enhanced renin release. These results indicate that insulin may play an important physiologic role in the renin-angiotensin system and suggest that proinsulin and amylin may be involved in the genesis and development of hypertension through enhancement of insulin-stimulated renin release. PMID- 11436179 TI - Sulfate could mediate the therapeutic effect of glucosamine sulfate. AB - Glucosamine sulfate is a controversial osteoarthritis remedy that is presumed to stimulate articular cartilage glycosaminoglycan synthesis by increasing glucosamine concentrations in the joint space. However, this is not plausible because even large oral doses of the product have no effect on serum glucosamine concentrations. We propose instead that sulfate could mediate the clinical benefit attributed to this treatment. Sulfate is required for glycosaminoglycan synthesis, and unlike glucosamine, its serum level can be modified by dietary and other factors. In this study, we tested whether oral glucosamine sulfate increases serum sulfate concentrations and whether the sulfate concentration in the synovial fluid reflects that in the serum. The serum sulfate concentration of 7 normal subjects was 331 +/- 21 micromol/L before ingestion of 1.0 g glucosamine sulfate and 375 +/- 17 micromol/L 3 hours after (P <.05). Serum sulfate concentrations decreased from 325 +/- 19 to 290 +/- 19 micromol/L when the same dose of glucosamine sulfate was ingested with 1.0 g of the analgesic drug acetaminophen, which is largely metabolized by sulfation (P <.05). Unlike glucosamine sulfate, oral sodium sulfate did not significantly increase the serum sulfate concentration. Synovial fluid and serum sulfate concentrations were closely similar when measured in 15 patients undergoing diagnostic needle aspiration of a knee effusion (r =.99, slope =.97, P <.0001). These results do not prove that glucosamine sulfate improves osteoarthritis, but considered with other data, they do provide a plausible biochemical mechanism for its reported beneficial effects. This hypothesis is clinically relevant because it predicts that nonsulfate salts of glucosamine will be ineffective and that renal function, diet, and concurrent acetaminophen therapy could confound clinical trials of this therapy. PMID- 11436180 TI - Altered GLUT1 and GLUT3 gene expression and subcellular redistribution of GLUT4: protein in muscle from patients with acanthosis nigricans and severe insulin resistance. AB - Multiple isoforms of glucose transporters are found in muscle, the tissue that normally accounts for 85% of insulin-stimulated glucose uptake. Glucose uptake into muscle cells in the fasting state is mediated primarily by GLUT1 and GLUT3 glucose transporters, whereas postprandial (insulin-stimulated) and exercise related increments in muscle glucose uptake are mediated primarily by GLUT4. To determine if glucose transporters are abnormally expressed in muscle from insulin resistant subjects, muscle samples were obtained from 10 normal subjects and 6 obese, nondiabetic subjects with severe insulin resistance and acanthosis nigricans. Both GLUT4 total protein and mRNA were normal in the insulin-resistant subjects. Muscle GLUT3 protein and mRNA were lower than controls by 62% and 71%, respectively. GLUT1 mRNA was twice normal, whereas GLUT1 protein content was not significantly increased. GLUT4 protein was markedly redistributed to the muscle plasma membrane in subjects with severe insulin resistance compared with normals (92% v 40% GLUT4 in plasma membrane-enriched fractions, P <.001), whereas the percentage of GLUT1 and GLUT3 protein found in the plasma membrane-enriched fractions was not different from controls. These data document differences in the expression of genes for GLUT1 and GLUT3 in muscle from normal and insulin resistant subjects. Further, insulin resistance with fasting hyperinsulinemia was associated with a redistribution of GLUT4 to the muscle cell surface with no change in total GLUT4 protein. These data suggest that glucose transporter gene expression and their basal distribution in human muscle are related to insulin resistance and could be determinants of whole body insulin responsiveness. PMID- 11436181 TI - Potassium and activity of the sodium hydrogen exchanger isoform 1 in vascular smooth muscle of hypertensive rats. AB - Low potassium intake is inversely associated with blood pressure. In vitro, the proliferation of vascular smooth muscle cells (VSMCs) shows an inverse correlation with [K(+)]. In hypertension, many studies have established that the ubiquitous Na(+)/H(+) exchanger isoform 1 (NHE-1) exhibits increased activity and is permissive for cell proliferation. Changes in extracellular [K(+)] lead to altered intracellular Na(+) content, which could affect NHE activity and NHE-1 protein expression. We therefore investigated the effects of altering extracellular [K(+)] on NHE activity and NHE-1 expression in cultured VSMCs of both the spontaneously hypertensive rat (SHR) and its normotensive Wistar-Kyoto counterpart (WKY). Culture of SHR VSMCs for 48 hours in media containing 2, 4, 6, and 8 mmol. L(-1) [K(+)] led to activation of NHE-1 in the low [K(+)] media (NHE 1 activity at [K(+)] 2, 4, 6, and 8 mmol. L(-1) were 34.3 +/- 1.7, 29.5 +/- 1.1, 27.7 +/- 1.4, and 26.1 +/- 2.1 mmol. L(-1) min(-1), P <.006 by analysis of variance [ANOVA]). This was not associated with any significant changes in intracellular pH. By contrast, WKY VSMCs did not exhibit any significant activation of NHE-1 in low [K(+)] media (NHE-1 activity at [K(+)] 2, 4, 6, and 8 mmol. L(-1) were 24.3 +/- 2.9, 22.3 +/- 1.7, 19.0 +/- 1.8, and 18.6 +/- 1.6 mmol. l(-1) min(-1), P = not significant [NS] by ANOVA). Culture of SHR or WKY VSMCs in low [K(+)] media did not alter NHE-1 protein expression, suggesting the enhancement of activity in SHR cells was due to an increased turnover number of NHE-1. This response of NHE-1 in SHR VSMCs to K(+) depletion indicated a direct effect on these cells and could potentially enhance the contractile or proliferative phenotype of these cells in vivo. PMID- 11436182 TI - Plasma cholesteryl ester transfer protein activity is not linked to insulin sensitivity. AB - Plasma cholesteryl ester transfer protein (CETP) activity has been reported to decline during a hyperinsulinemic-euglycemic clamp. It has been suggested that this suppressive effect of acute hyperinsulinemia is linked to whole body insulin sensitivity, and that the insulin resistance that accompanies obesity leads to high plasma CETP activity found in obese subjects. In the present study, we used 2 experimental approaches to examine the putative link between CETP and insulin action. First, we examined if the clamp-induced suppression of plasma CETP activity is linked to whole body insulin sensitivity. Plasma CETP activity was measured at the beginning and end of a 2-hour hyperinsulinemic-euglycemic clamp in 18 nondiabetic individuals before and after an exercise training regimen that improved insulin sensitivity without weight loss. CETP activity decreased in response to the clamp procedure in 16 of 18 subjects, and on average, by 9% (P <.001). While training decreased plasma CETP activity (10%, P <.05), the improvement in insulin sensitivity had no statistical effect on the clamp-induced suppression of plasma CETP activity (training*clamp, P =.26). Second, we examined if insulin resistance is associated with an elevation in fasting plasma CETP activity when the influence of adiposity and diabetes were negated. Plasma CETP activity was measured in 41 women (12 insulin-sensitive lean; 8 insulin-resistant lean; 10 insulin-sensitive obese; 11 insulin-resistant obese). The level of insulin sensitivity had no significant effect on fasting plasma CETP activity, but CETP levels were 25% higher in obese subjects (P <.01). Thus, neither experimental approach provided evidence that plasma CETP levels are linked to insulin and insulin sensitivity. These data suggest that the elevated CETP activity found in obese patients is less associated with hyperinsulinemia and the accompanying insulin resistance, but rather is more related to some other metabolic complication of obesity. PMID- 11436183 TI - High glucose stimulates hyaluronan production by renal interstitial fibroblasts through the protein kinase C and transforming growth factor-beta cascade. AB - Deposition of hyaluronan has been implicated in the pathogenesis of diabetic glomerulosclerosis. We hypothesized the involvement of hyaluronan in diabetic tubulointerstitial fibrosis. We investigated high-glucose effect on hyaluronan production by rat renal interstitial fibroblasts (normal rat kidney [NRK] cells) and examined the role of hyaluronan in NRK cell proliferation. The involvement of protein kinase C (PKC) and transforming growth factor-beta (TGF-beta) in this response was also examined. After 24 hours of incubation in medium containing 25.6 mmol/L glucose, production of hyaluronan by NRK cells was significantly increased compared with medium containing 5.6 mmol/L glucose (P <.01). L-glucose and mannitol had no effect on hyaluronan production. High glucose enhanced basal in situ PKC activity (P <.01), and both an activator of PKC (phorbol 12-myristate 13-acetate; [PMA]) and TGF-beta 1 were able to increase hyaluronan production by NRK cells (P <.01). The effect of high glucose on hyaluronan production was diminished by coincubating cells with PKC inhibitors (staurosporine [Stp] or calphostin C [CpC]) or with an anti-TGF-beta neutralizing antibody. Stimulation of hyaluronan production by PMA was also normalized by anti TGFbeta neutralizing antibody, but the effect of TGF-beta1 was not affected by inhibition of PKC. Finally, incubating quiescent NRK cells with 50 or 100 ng/mL hyaluronan for 24 hours significantly increased NRK cell number (P <.01). In conclusion, high glucose stimulates hyaluronan production through the PKC/TGF-beta cascade. Increased hyaluronan promotes NRK cell proliferation. These results suggest that hyaluronan may play a role in the pathogenesis of interstitial fibrosis in diabetic kidney disease. PMID- 11436184 TI - Relationship between insulin resistance, weight loss, and coronary heart disease risk in healthy, obese women. AB - Several popular books have recently been published stating that being insulin resistant favors weight gain and/or prevents weight loss. Because this view seems to have gained widespread support in the general population, we thought it important to perform the current study testing the hypothesis that differences in insulin-mediated glucose disposal do not affect weight loss in response to calorie-restricted diets. For this purpose, we studied the change in weight and risk factors for coronary heart disease (CHD) in healthy women volunteers, defined as being obese on the basis of a body mass index (BMI) greater than 30.0 kg/m(2). The insulin suppression test was used to stratify obese women at baseline into insulin-resistant and insulin-sensitive subgroups on the basis of their steady-state plasma glucose (SSPG) concentration at the end of a 180-minute infusion of octreotide, exogenous insulin, and glucose. They were then instructed on a calorie-restricted diet plus sibutraminine (15 mg/day) for a total period of 4 months. Baseline measurements also included determination of fasting lipid and lipoprotein concentrations, and hourly (8 AM to 4 PM) determinations of plasma glucose and insulin concentrations before and after breakfast and lunch. Twenty four women completed the 4-month period of calorie restriction: 13 classified as insulin-resistant (SSPG = 219 +/- 7 mg/dL) and 11 as insulin-sensitive (SSPG = 69 +/- 6 mg/dL). The insulin-resistant group also had higher (P =.03) plasma triglyceride (TG) concentrations and a higher ratio of total to high-density lipoprotein (HDL) cholesterol concentration (P =.02) at baseline. Both groups lost a significant amount of weight during the study, and there was no difference between the weight loss in the insulin-resistant (8.6 +/- 1.3 kg) and insulin sensitive (7.9 +/- 1.4 kg) groups. Weight loss in the insulin-resistant group was also associated with a significant decrease in SSPG concentration (219 +/- 7 to 144 +/- 14 mg/dL), associated with significantly lower fasting TG concentrations (P <.001) and day-long concentrations of plasma glucose and insulin (P <.005). None of these variables changed in the insulin-sensitive group. These results indicate that: (1) CHD risk factors in obese women vary as a function of being insulin-resistant or insulin-sensitive; (2) dramatic variations in insulin mediated glucose disposal do not modulate weight loss in response to calorie restricted diets, and (3) weight loss is effective in reducing CHD risk in insulin-resistant, obese women. Given these data, it seems obvious that attempts to reduce CHD risk factors by weight loss should focus on obese individuals who are also insulin-resistant. PMID- 11436185 TI - Assessing systemic 11beta-hydroxysteroid dehydrogenase with serum cortisone/cortisol ratios in healthy subjects and patients with diabetes mellitus and chronic renal failure. AB - 11beta-hydroxysteroid dehydrogenase (11beta-HSD), an enzyme regulating mineralocorticoid like action of glucocorticoid, oxidizes active cortisol to inactive cortisone. Impaired activity of this enzyme is associated with apparent mineralocorticoid excess (AME) syndrome and is characterized by hypertension and hypokalemia. Recent investigations suggest the presence of hypertensive subjects with low activity of 11beta-HSD. The blood concentration ratio of cortisone/cortisol reflects the overall conversion of cortisol to cortisone and may be an index to assess the systemic activity of 11beta-HSD. We evaluated the peripheral blood concentration ratio of cortisone/cortisol as a possible marker to identify subjects with hypertension thought to represent impaired 11beta-HSD activity. We compared this ratio in healthy subjects and patients with diabetes mellitus (DM) or chronic renal failure (CRF). Peripheral blood samples were collected from 69 healthy subjects, 44 DM, and 36 CRF patients in the morning (9:00 to 11:00 AM). Twenty-six DM patients (59%) and 32 CRF patients (89%) met the criteria for having hypertension. Serum cortisol and cortisone concentrations were determined by high performance liquid chromatography (HPLC). All values for serum cortisone and cortisol levels were within the normal range. Serum cortisone/cortisol ratio in the healthy subjects was distributed with a range of 0.113 to 0.494 (median, 0.243). Compared with healthy subjects, DM and CRF patients had significantly low (P <.01) serum cortisone/cortisol levels (median, 0.188 [range, 0.092 to 0.313] in DM and 0.088 [range, 0.031 to 0.140] in CRF). Bimodal distribution of cortisone/cortisol, found in DM patients with hypertension, represented high- and low-ratio groups around the border of the ratio 0.2. Kidney function, DM duration, and complications varied between the high- and low-ratio groups. The low ratio group (<0.2), whose 11beta-HSD activity was considered low, had an increase in blood urea nitrogen (BUN) levels and experienced nephropathy, neuropathy, retinopathy, and prolonged DM duration when compared with the group with a ratio greater than 0.2. The data suggest that the serum cortisone/cortisol ratio reflects the change in 11beta-HSD activity and is dependent kidney function. This is a possible marker to evaluate glucocorticoid excess hypertension observed in DM and CRF patients. PMID- 11436186 TI - Effect of insulin resistance on serum paraoxonase activity in a nondiabetic population. AB - Paraoxonase is a high-density lipoprotein (HDL)-bound esterase that hydrolyzes various organophosphorus compounds and protects low-density lipoprotein (LDL) against accumulation of lipid peroxides. Paraoxonase activity is strongly affected by the polymorphism of the paraoxonase gene (PON1) at position 192. In addition, the enzyme activity shows a great variation within each genotype, although the underlying mechanism is unknown. Because paraoxonase activity is decreased in subjects with type 2 diabetes mellitus who have insulin resistance, we investigated the association between paraoxonase activity and insulin resistance in a nondiabetic population. The subjects were 237 healthy Japanese adults with fasting plasma glucose less than 7.0 mmol/L. Paraoxonase activity was measured using paraoxon as a routine substrate. Insulin resistance was assessed by homeostasis model assessment index (HOMA index). Paraoxonase activity was affected by HDL level. To reduce the effect of HDL on paraoxonase, paraoxonase activity/HDL ratio was used. When the subjects were divided into tertiles by HOMA index, the subjects with higher HOMA values had higher paraoxonase/HDL ratios, although the 3 groups were comparable in age, gender and the PON1 genotype distribution. Paraoxonase/HDL ratio showed significant positive correlations not only with HOMA index, but also with body mass index, waist-to-hip ratio (WHR), whereas it correlated inversely with age at borderline significance. Multiple regression analysis indicated that the association between HOMA index and paraoxonase/HDL ratio was significant and independent of PON1 genotype, age, and adipocity. The positive association between HOMA index and HDL-corrected enzyme activity was again significant when the enzyme activity was measured with diazoxon as an alternative substrate. These results suggest that insulin resistance or hyperinsulinemia is a factor contributing to the intragenotype variability of paraoxonase activity in a population without overt hyperglycemia. PMID- 11436187 TI - Pathophysiologic phenotypes of Japanese subjects with varying degrees of glucose tolerance: using the combination of C-peptide secretion rate and minimal model analysis. AB - We tried to characterize the clinical features associated with glucose metabolism in the development of diabetes. Study subjects were glucose-tolerant subjects without a family history of diabetes (normal glucose tolerance [NGT]1 group, n = 15) and with a first-degree diabetes relative (NGT2, n = 9), 12 subjects with impaired glucose tolerance (IGT), and 13 subjects with type 2 diabetes mellitus (DM). The first phase C-peptide secretion (CS1), insulin sensitivity (Si), and glucose effectiveness (Sg) were assessed by the combination of C-peptide 2 compartment model and minimal model analyses. Using these parameters, each group was characterized: CS1 was decreased in NGT2 and IGT compared with NGT1 and further decreased in DM; Si was not different among NGT1, NGT2, and IGT, whereas Si was decreased in DM; CS1 x Si value was decreased in NGT2 compared with NGT1 and decreased in IGT, DM, progressively; Sg was decreased in IGT and DM compared with NGT1 and NGT2. CS1 x Si and Sg values could segregate each group distinctively, although it had a large variety of phenotypes. CS1 x Si value and Sg are assumed to represent the contributions of insulin-dependent and independent mechanisms to glucose tolerance, respectively, and thus, both mechanisms should play an important role in the characterization of pathophysiologic phenotypes of the subjects with various degrees of glucose tolerance. PMID- 11436188 TI - Metabolic changes following sibutramine-assisted weight loss in obese individuals: role of plasma free fatty acids in the insulin resistance of obesity. AB - The relationship between insulin-mediated glucose disposal and daylong free fatty acid (FFA) concentrations before and after sibutramine-assisted weight loss was investigated in 24 healthy, normotensive, nondiabetic, obese women (body mass index [BMI] >30.0 kg/m(2)). The 24 volunteers were defined as being insulin resistant (IR) or insulin-sensitive (IS) on the basis of their steady-state plasma glucose (SSPG) concentration in response to a 180-minute continuous intravenous infusion of octreotide, insulin, and glucose. The mean (+/- SEM) SSPG concentrations were significantly higher (P <.001) in the IR group (219 +/- 7 v 69 +/- 6 mg/dL) at baseline. The IR group also had significantly higher plasma glucose (P =.002), insulin (P <.001), and FFA (P =.02) concentrations measured at hourly intervals from 8 AM to 4 PM, before and after breakfast (8 AM) and lunch (noon). Weight loss in response to an energy-restricted diet for 4 months and sibutramine (15 mg/d) was comparable in the 2 experimental groups (8.6 +/- 1.3 v 7.9 +/- 1.4 kg). SSPG concentrations decreased significantly (P <.001) following weight loss (219 +/- 7 to 144 +/- mg/dL) in the IR group, but there was no change in the SSPG of the IS group (69 +/- 6 to 73 +/- 7 mg/dL. The improvement in insulin sensitivity in the IR group after weight loss was associated with a significant decline in daylong plasma glucose (P >.001) and insulin (P =.02) concentrations, without a weight-loss-associated decrease in daylong plasma FFA responses. In contrast, there was no significant change in plasma glucose, insulin, and FFA concentrations following weight loss in the IS group. These results indicate that daylong FFA concentrations vary substantially in obese individuals as a function of whether they are IR or IS. Furthermore the observation that the IR group was more insulin-sensitive after weight loss, associated with lower daylong insulin concentrations in the absence of a significant decrease in circulating FFA concentrations, suggests that resistance to insulin-mediated glucose disposal in obese individuals cannot be entirely due to high FFA levels. PMID- 11436189 TI - Correlation between carotid artery distensibility and serum vascular endothelial growth factor concentrations in type 1 diabetic subjects and nondiabetic subjects. AB - The relationships between serum vascular endothelial growth factor (VEGF) concentrations and vessel wall ultrasonic characteristics in type 1 diabetic and nondiabetic subjects were assessed. Serum VEGF concentration was measured, and ultrasound imaging and blood pressure recordings were performed in 41 type 1 diabetic subjects (hemoglobin A(1c) [HbA(1c)], 7.63 [1.17%]; duration of diabetes, 12 (0 to 23) years), and 50 nondiabetic subjects. Change in carotid artery luminal diameter during the cardiac cycle was measured using M-mode ultrasound, from which percentage increase in carotid artery luminal diameter was calculated; the carotid artery distensibility index was calculated as the ratio of percentage increase in carotid artery luminal diameter and pulse pressure. Serum VEGF concentration was higher in the diabetic subjects (217 [135 to 336] v 137 [80 to 237] pg/mL; P =.009). The percentage increase in carotid luminal diameter during the cardiac cycle was not significantly different between the 2 groups (12.9 [10.2 to 15.7] v 13.0 [10.6 to 15.0%]; P =.270) despite significantly greater pulse pressure in the type 1 diabetic group (55 [45 to 71] v 46 [41 to 51] mm Hg; P =.0003). The distensibility index was therefore lower in the diabetic subjects (0.24 [0.10] v 0.28 [0.08%]/mm Hg; P =.031). There was a significant negative correlation between serum VEGF concentrations and mean percentage increase in carotid luminal diameter during the cardiac cycle in the diabetic group (r = -.36, P =.021) and in the nondiabetic group (r = -.28, P =.047). This negative correlation could be strengthened by relating mean percentage increase in luminal diameter to pulse pressure to give the distensibility index. Therefore, serum VEGF concentrations correlated strongly and inversely with the distensibility index in the diabetic group (r = -.49, P =.001), in the nondiabetic group (r = -.29, P =.041), and in both groups analyzed together (r = -.42, P <.0001). Vessel wall distensibility may be an important determinant of serum VEGF concentrations in both diabetic and nondiabetic populations and may underlie the previously observed association between blood pressure and serum VEGF concentrations. The pathophysiologic relevance of these findings remains to be elucidated. PMID- 11436190 TI - Retinoid X receptor expression in skeletal muscle of nondiabetic, obese and type 2 diabetic individuals. AB - Retinoid X receptor (RXR) is a nuclear receptor that functions as an obligate heterodimeric partner of peroxisome proliferator-activator receptor (PPAR). Studies have shown that the alpha isoform of RXR and PPARgamma act synergistically to regulate gene expression and insulin action. The aim of the current study was to compare the expression and regulation of RXR in the primary insulin-sensitive tissue, skeletal muscle, of various degrees of insulin resistant states including obese type 2 diabetic (T2D), obese nondiabetic (OND), and lean nondiabetic (LND) subjects. Insulin action/resistance was determined by a 3-hour hyperinsulinemic, euglycemic (5.0 to 5.5 mmol/L) clamp. Percutaneous biopsy of the vastus lateralis muscle was performed before and after the clamp. RXRalpha mRNA was measured using a quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) assay, while protein was determined by Western blotting. All 3 isoforms of RXR, alpha, beta, and gamma, were present in skeletal muscle. Protein expression of RXR isoforms did not differ between groups; RXR alpha mRNA was also similar between groups. Neither RXR alpha mRNA, RXR -beta nor -gamma protein displayed significant relationships with any of the clinical or laboratory parameters measured, including insulin sensitivity. RXR alpha exhibited a negative correlation with free fatty acids levels (r, -.42, P <.05). There was also no relationship between RXR alpha and PPARgamma protein levels. RXR alpha mRNA was unaltered following insulin infusion. We conclude that RXR isoform (alpha, beta, gamma) expression is not tightly controlled by insulin, insulin resistance or type 2 diabetes. Instead, RXR isoforms are likely constitutive proteins or controlled by other factors. PMID- 11436191 TI - Relationship between hormone replacement therapy use with body fat distribution and insulin sensitivity in obese postmenopausal women. AB - The aim of this study was to compare the effect of hormone replacement therapy (HRT) on insulin resistance and central adiposity in obese postmenopausal women. Forty-five obese postmenopausal women (16 HRT users and 29 nonusers), with a mean age of 56.6 +/- 5.3 years and duration of current, continuous HRT use of 4.7 +/- 2.9 years, were included in the study. Subjects were studied using oral glucose tolerance tests, euglycemic clamping, dual photon x-ray absorptiometry, computed tomography, doubly labeled water, and treadmill testing. Insulin sensitivity, total fat, visceral fat, subcutaneous abdominal fat, thigh muscle attenuation, daily physical activity energy expenditure, peak oxygen consumption (Vo(2)) were measured. HRT users had lower body weight (88.0 +/- 11.0 v 98.2 +/- 15.0 kg, P =.05), lower body mass index (33.1 +/- 3.5 v 36.8 +/- 5.2 kg/m(2), P =.05), lower fat mass (38.3 +/- 7.3 v 44.1 +/- 10 kg, P =.05), less visceral adipose tissue (157 +/- 47 v 211 +/- 81 cm(2); P =.05), and higher peak Vo(2) (21.1 +/- 4.6 v 17.6 +/- 2.2 mL/kg/min, P =.001) than nonusers. After adjustment for total fat, we noted a trend for decreased visceral adipose tissue in HRT users (P =.09). After adjustment for peak Vo(2), the decreased visceral adipose tissue persisted in HRT users (P <.01). Insulin sensitivity per kilogram of lean body mass did not differ between HRT users (0.51 +/- 0.22 mmol/kg/min) and nonusers (0.49 +/- 0.22 mmol/kg/min). It was concluded that obese postmenopausal women using HRT have a more favorable body composition and fat distribution pattern than nonusers. Although visceral adipose tissue is decreased in HRT users, insulin sensitivity does not differ between HRT users and nonusers. PMID- 11436192 TI - Changes in blood lipids consequent to aerobic exercise training related to changes in body fatness and aerobic fitness. AB - The contribution of changes in body fatness and aerobic fitness to changes in blood lipids after aerobic exercise training was investigated. The sample included 295 men (77 black, 218 white) and 355 women (131 black, 224 white), aged 17 to 65 years, from the HERITAGE Family Study. Participants underwent measurements at baseline and after 20 weeks of supervised exercise training on a cycle ergometer. Body fat mass (FM, in kilograms) was determined by underwater weighing, and aerobic fitness (maximal oxygen uptake, VO(2max), in milliliters per minute) was assessed by cycle ergometry. Blood lipid measurements included fasting plasma levels of high-density lipoprotein cholesterol (HDL-C), HDL(2)-C, HDL(3)-C, low-density lipoprotein cholesterol (LDL-C), total cholesterol (CHOL), CHOL/HDL, and triglycerides (TG). A composite lipid change index (LCI) was derived by subjecting the Delta scores for the individual blood lipids to principal components analysis. The exercise training was accompanied by a mean increase of 17.5% in VO(2max) and a mean decrease of 3.3% in FM. Partial correlations, controlled for age, between absolute changes in VO(2max) and changes in the blood lipids were consistently low and nonsignificant. On the other hand, absolute changes in FM were significantly (P <.05) associated with changes in HDL-C (r = -.23), HDL(2)-C (r = -.17), and CHOL/HDL (r =.24) and the LCI (r = -.27) in men and with changes in LDL-C (r =.22), CHOL (r =.19), and CHOL/HDL (r =.15) and the LCI (r = -.19) in women. Forward stepwise regression confirmed that the change in FM was a better predictor of changes in blood lipids than the change in VO(2max), entering as a predictor in 4 of 8 regressions in both men and women. Change in VO(2max) did not enter as a significant predictor in any regression. Further, there were no differences in LCI between the upper and lower quartiles of VO(2max) change. On the other hand, there were significant differences between the low and high quartiles of FM change. No race effects were observed in any of the relationships, except that race was a significant predictor of changes in TG in both men and women. In conclusion, changes in blood lipids associated with aerobic exercise training do not appear to be related to changes in aerobic fitness per se; rather, they are weakly to moderately associated with changes in body fatness. PMID- 11436193 TI - Effects of moderate and high glycemic index meals on metabolism and exercise performance. AB - The purpose of this study was to determine whether pre-exercise ingestion of meals with moderate and high glycemic indexes (GI) affects glucose availability during exercise and exercise performance time. Six male volunteers (22 +/- 1 years; 80.4 +/- 3.7 kg; VO(2peak), 54.3 +/- 1.2 ml. kg(-1). min(-1)) ingested 75 g of carbohydrate in the form of 2 different breakfast cereals, rolled oats (moderate GI, approximately 61; MOD-GI) or puffed rice (high GI, approximately 82; HI-GI), combined with 300 mL of water; or water alone (control). The trials were randomized, and the meals were ingested 45 minutes before the subjects performed cycling exercise (60% VO(2peak)) to exhaustion. Venous blood samples were drawn to measure glucose, free fatty acids (FFAs), glycerol, insulin (INS), epinephrine (EPI) and norepinephrine (NE) concentrations. A muscle biopsy specimen was obtained from the vastus lateralis before the meal and immediately after exercise for glycogen determination. Before exercise, both test meals elicited significant (P <.05) hyperglycemia and hyperinsulinemia compared with control. The glycemic response was higher (P <.05) at the start of exercise after the HI-GI meal than after the control. During exercise, plasma glucose levels were higher (P <.05) at 60 (5.2 +/- 0.1, 4.2 +/- 0.2, and 4.6 +/- 0.1 mmol. L( 1)) and 90 (4.8 +/- 0.1, 4.1 +/- 0.1, and 4.3 +/- 0.1 mmol. L(-1)) minutes after the MOD-GI meal than after either the HI-GI or control. Total carbohydrate oxidation was greater (P <.05) during the MOD-GI trial than in control and was directly correlated with exercise performance time (r =.95, P <.0001). Pre exercise plasma FFA levels were suppressed (P <.05) 30 and 45 minutes after ingestion of the HI-GI meal and 45 minutes after the MOD-GI meal compared with control. At 30, 60, and 120 minutes of exercise, FFAs remained suppressed (P <.05) for both test meals compared with control. At exhaustion, plasma glucose, INS, FFA, glycerol, EPI, and NE levels and muscle glycogen use were not different for all trials. Exercise time was prolonged (P <.05) after the MOD-GI meal compared with control, but the HI-GI trial was not different from control (MOD GI, 165 +/- 11; HI-GI, 141 +/- 8; control, 134 +/- 13 minutes). Thus, in contrast to the HI-GI meal or control, the MOD-GI breakfast cereal ingested 45 minutes before exercise enhanced performance time, maintained euglycemia for a longer period during exercise, and resulted in greater total carbohydrate oxidation during the exercise bout. We conclude that a MOD-GI meal provides a significant performance and metabolic advantage when consumed 45 minutes before exercise. PMID- 11436194 TI - Metformin reduces weight, centripetal obesity, insulin, leptin, and low-density lipoprotein cholesterol in nondiabetic, morbidly obese subjects with body mass index greater than 30. AB - We studied 31 nondiabetic, habitually (> or =5 years) morbidly obese subjects (mean +/- SD body mass index [BMI] 43 +/- 8.7, median 43). Our specific aim was to determine whether metformin (2.55 g/d for 28 weeks) would ameliorate morbid obesity and reduce centripetal obesity; lipid and lipoprotein cholesterol, insulin, and leptin levels; and plasminogen activator inhibitor activity (PAI Fx), risk factors for coronary heart disease (CHD). The patients were instructed to continue their prestudy dietary and exercise regimens without change. After 2 baseline visits 1 week apart, the 27 women and 4 men began receiving metformin, 2.55 g/d, which was continued for 28 weeks with follow-up visits at study weeks 5, 13, 21, and 29. Daily food intake was recorded by patients for 7 days before visits then reviewed with a dietitian. Kilocalories per day and per week were calculated. At each visit, fasting blood was obtained for measurement of lipid profile, insulin, leptin, and PAI-Fx. The mean +/- SD kilocalories consumed per day, 1,951 +/- 661 at entry, fell by week 29 to 1,719 +/- 493 (P =.014) but did not differ at weeks 5, 13, and 21 from that at week 29 (P >.2). Weight fell from 258 +/- 62 pounds at entry to 245 +/- 54 pounds at week 29 (P =.0001). Girth was reduced from 51.8 +/- 6.2 to 49.2 +/- 4.5 inches (P =.0001). Waist circumference fell from 44.0 +/- 6.4 inches to 41.3 +/- 5.9 (P =.0001). The waist/hip ratio fell from 0.85 +/- 0.09 to 0.84 +/- 0.09 (P =.04). Fasting serum insulin, 28 +/- 15 microU/mL at entry, fell to 21 +/- 11 microU/mL at week 29 (P =.0001), and leptin fell from 79 +/- 33 ng/mL to 55 +/- 27 ng/mL (P =.0001). On metformin, there were linear trends in decrements in weight, girth, waist circumference, waist/hip ratio, insulin, and leptin throughout the study period (P <.007). Low density lipoprotein (LDL) cholesterol, 126 +/- 34 mg/dL at study entry, fell to 112 +/- 43 mg/dL at week 29 (P =.001), with a linear trend toward decreasing levels throughout (P =.036). By stepwise linear regression, the higher the entry weight, the larger the reduction in weight on metformin therapy (partial R(2) = 31%, P =.001). The greater the reduction in kilocalories consumed per day, the greater the decrease in weight on metformin therapy (partial R(2) = 15%, P =.011). The higher the waist/hip ratio at entry, the greater its reduction on metformin therapy (partial R(2) = 11%, P =.004). The higher the entry serum leptin, the greater its reduction on metformin therapy (partial R(2) = 29%, P =.002). The greater the reduction in insulin on metformin, the greater the reduction in leptin (partial R(2) = 8%, P =.03). The higher the entry PAI-Fx, the greater the reduction in PAI-Fx on metformin (partial R(2) = 43%, P =.0001). Metformin safely and effectively reduces CHD risk factors (weight, fasting insulin, leptin, LDL cholesterol, centripetal obesity) in morbidly obese, nondiabetic subjects with BMI > 30, probably by virtue of its insulin-sensitizing action. PMID- 11436195 TI - Application of an improved biuret method to the determination of total protein in urine and cerebrospinal fluid without concentration step by use of Hitachi 7170 auto-analyzer. AB - A biuret automated colorimetric assay for total protein in urine and cerebrospinal fluids was established. The procedures were as follows. Acidify all urine sample before analysis. Add precipitant Na(2)WO(4) to urine samples. After 10 min, centrifuge, decant the supernatant fluid, drain the inverted tubes on absorbent tissue, dissolve the precipitation with 0.1 mol/L NaOH, and finally adapt the reconstituted urine to the Hitachi 7170 analyzer. A cell-free cerebrospinal fluid sample produced by centrifugation can be inserted in an auto analyzer for protein measurement directly. The program: mix 35 microl sample (CSF or reconstituted urine) and standard with 0.2 mol/L NaOH; incurable at 37 degrees C for 5 min, and real A1. Add concentrated biuret reagent, and 10 min later measure absorbance A2 at 546 nm vs. reagent blank. Secondary wavelength was 700 nm. The test results were calculated against a one-point standard. This biuret colorimetric method was relatively simple, fast, and accurate for the determination of protein in urine and cerebrospinal fluid, with a wide linearity extending from 0.125 g/L up to 6 g/L, had a good correlation with Benzethonium chloride turbidimetry technique, and was a practical routine method. PMID- 11436196 TI - Sensitive and rapid immunoassays for Salmonella enteritidis. AB - In order to develop more sensitive, specific, and rapid immunoassays to detect Salmonella enteritidis in food supplies, we have applied various approaches by using several different antibody preparations. Utilizing ELISA in both a plate and immunodot assay, we employed (i) a polycolonal rabbit antiserum to a boiled suspension of the organism; (ii) a monoclonal antibody to the cell surface of the bacterium; and (iii) mouse antisera to two oligosaccharides each containing the rare sugar tyvelose, and exhibited by S. enteritidis, a member of the group D salmonellae. We showed that the polyclonal antiserum and monoclonal antibody IG 10 to the cell surface could specifically detect from 10(2) to 10(3) organisms in a 10-microl sample in the plate and immunodot assay. Both assays are read in 4-5 hr. Further, in the mice immunized to the trisaccharide, (alpha-D-galactose-alpha tyvelose-alpha-D-mannose), as well as those mice immunized to the tetrasaccharide, (alpha-D-galactose-alpha-tyvelose-alpha-D-mannose-alpha-L rhamnose), specificity to tyvelose was determined by inhibition studies. The inhibitors of the antisera to the trisaccharide included the single sugar tyvelose conjugated to bovine serum albumin (BSA), a tetrasaccharide in which tyvelose is excluded, but contains alpha-D-galactose, alpha-ascarose, alpha-D mannose, and alpha-L-rhamnose (conjugated to BSA), and others. The inhibition studies suggest that the mouse antisera are specific for tyvelose and also contain antibodies for mannose and rhamnose. The antibodies that have been made to the unique sugar tyvelose should improve the specificity in assays for S. enteritidis. PMID- 11436197 TI - Semiquantitative analysis of urinary low protein levels using silver dot blot assay. AB - We designed a semiquantitative analysis of urinary low protein levels using silver dot blot assay. In this method, 3 microl of urine are blotted to one dot onto a cellulose acetate membrane, which is stained by a colloidal silver staining reagent, and the optical density of the silver stained urinary protein is measured at 500 nm using a densitometer. There was a good linearity between 2.5 mg/L and 100 mg/L and a gentle linearity between 100 mg/L and 200 mg/L, and the minimum sensitivity was 2.5 mg/L. This method is suitable for semiquantitative analysis of urinary protein levels less than 300 mg/L, which can not be determined precisely by dipstick. PMID- 11436198 TI - Effect of elevated concentration of alkaline phosphatase on cardiac troponin I assays. AB - Troponin I is the regulatory subunit of troponin complex associated with the actin thin filament within muscle cells. Cardiac troponin I (cTnI) is a good marker for diagnosis of myocardial damage. Several immunoassays are available for determination of cTnI in serum. The Stratus cTnI fluorometric enzyme immunoassay (Dade International) uses alkaline phosphatase (ALP) substrate. The microparticle enzyme immunoassay (MEIA) for cTnI (Abbott Laboratories) also uses ALP conjugate. On the other hand, the chemiluminescent assay (CLIA) for cTnI (Bayer Diagnostics) does not use ALP. ALP activity may frequently be elevated in serum of patients being evaluated for suspected myocardial infarction. Therefore, we studied the potential interference of ALP in cTnI assays. Serum pools were prepared from patients, and various concentrations of ALP solution were added to different aliquots. The cTnI concentrations were measured by the Stratus, MEIA, and CLIA assays. We observed no interference of ALP in the MEIA and CLIA assay for cTnI. On the other hand, we observed significant positive interference of ALP when cTnI concentrations were measured using the Stratus. PMID- 11436199 TI - Electrophoretic analysis of urinary proteins in diabetic adolescents. AB - Pathological changes in the urine sodium dodecyl sulphate gel electrophoresis (SDS PAGE) patterns often precede the occurrence of any sign of renal involvement in diabetes. However, data concerning the most frequent SDS PAGE pattern of the urine in early stages of type I diabetes mellitus are controversial. In the present study an SDS PAGE technique has been used that provides an adequate sensitivity for the detection of the abnormal pattern. Urinary proteins have been analyzed by SDS PAGE in twenty two diabetic adolescents and twenty four age matched controls. Albumin concentration, and N acetyl-beta-D-glucosaminidase (NAG) activity were also measured in the same samples. There was no significant difference in urine albumin concentration and NAG activity between diabetic children and controls. However twelve patients showed an electrophoretic pattern characteristic for glomerulopathy, two had a pattern indicating tubular dysfunction and another two patients had a mixed pattern. Among the twenty four controls only three showed abnormal electrophoretic patterns. The results support the view that early stages of diabetic nephropathy may involve both glomerular and tubular dysfunction. However the exact clinical and prognostic significance of the information provided by SDS PAGE analysis remains to be elucidated. PMID- 11436200 TI - p21 gene codon 31 arginine/serine polymorphism: non-association with endometriosis. AB - p21, an important regulator of the cell cycle, acts as a mediator of the growth suppressing and -promoting functions of p53. We aimed to investigate the association between codon 31 polymorphisms of p21 gene and endometriosis. Women were divided into two groups: endometriosis (n = 102) and nonendometriosis (n = 119). The gene polymorphism for p21 codon 31 involved a base change from AGC to AGA and amino acid changes from serine (Ser) to arginine (Arg). Polymorphisms (Ser homozygotes, heterozygotes, Arg homozygotes) between both groups were detected and compared. Associations between the endometriosis and polymorphisms were evaluated. The results revealed that the distributions of different p21 polymorphisms in both groups were nonsignificantly different. The proportions of Ser homozygote/heterozygote/Arg homozygote in endometriosis and nonendometriois populations were 26.5/48.0/25.5% and 17.6/50.4/31.9%, respectively. We concluded the noncorrelation between the endometriosis and the p21 codon 31 polymorphism. p21 gene codon 31 arginine/serine polymorphism is not a useful marker for prediction of endometriosis susceptibility. PMID- 11436201 TI - Asian multicenter trials on urinary type IV collagen in patients with diabetic nephropathy. AB - To investigate the changes of renal type IV collagen turnover in diabetic nephropathy, urinary type IV collagen was measured by a highly sensitive one-step sandwich enzyme immunoassay (EIA). Urinary samples were obtained from 698 diabetic patients and 191 healthy adults. Among the patients, 264 had urinary albumin levels of less than 29 mg/g.creatine (Cr) (Stage I: normoalbuminuric stage), 169 had microalbuminuria from 30 to 299 mg/g.Cr (Stage II: microalbuminuric stage), 84 patients had macroalbuminuria of more than 300 mg/g.Cr and serum Cr of less than 1.1 mg/dl (Stage IIIA: macroalbuminuric stage without renal dysfunction), 97 had macroalbuminuria of more than 300 mg/g.Cr and serum Cr of more than 1.2 mg/dl (Stage IIIB: macroalbuminuric stage with renal dysfunction), and 84 had renal failure (Stage IV). The levels of urinary type IV collagen in Stages II, IIIA, IIIB, and IV were significantly higher than those in Stage I (P < 0.0001). The level of urinary type IV collagen in Stage I (5.00 +/- 0.23 microg/g.Cr; mean +/- SE) was also higher than that in normal adults (3.44 +/- 0.11 microg/g.Cr; mean +/- SE). These levels increased gradually due to progression of the clinical stage of diabetic nephropathy. It appears that the levels of urinary type IV collagen can be a useful marker for detecting renal injuries in diabetes according to our Asian multicenter trials. PMID- 11436202 TI - Characterization of serum and urinary chromogranin A by size exclusion chromatography: impact on calibrator selection and urinary assay. AB - Serum chromogranin A (CgA) is a useful marker for neuroendocrine tumors and is detectable in carcinomas at advanced stages. Elevated serum CgA is also an indicator of poor prognosis in prostate cancer and is useful for predicting the failure of hormonal therapy for prostate cancer patients. We found that CgA molecules with three different sizes could be detected in normal human serum. However, only the largest CgA molecule appears in patients with liver disease. Serum taken from cancer patients is composed predominantly of the middle-sized molecule, whereas the smallest CgA molecule was elevated in serum drawn from renal patients. Moreover, only the smallest CgA molecule was found in urine. We believe that the largest CgA molecule is metabolized by the liver, whereas the smallest CgA molecule is removed from the blood circulation via the kidney. Because the medium-sized CgA is the dominant molecule in both the cell medium of the tumor cell line SK-N-AS and sera from patients with malignant diseases, CgA from the cell medium was selected as the calibrator for the CgA ELISA assay. Our findings also suggest that it would not be possible to measure the urinary CgA to reflect the serum CgA concentration in order to detect pheochromocytoma among patients with hypertension. PMID- 11436203 TI - Semi-quantitative analysis of cytokine mRNA expression induced by the herbal medicine Sho-saiko-to (TJ-9) using a Gel Doc system. AB - The RT-PCR method was employed to determine the cytokine mRNA expression of human peripheral lymphocytes induced by the Japanese herbal medicine Sho-saiko-to (TJ 9). The results showed that the mRNA expression of IL-12, IL-1beta, IL-10, TNF alpha, G-CSF, and IFN-gamma increased after 6 hr in culture. This is the first reported finding that TJ-9 is an IFN-gamma inducer. Next, cytokine mRNA expression was semi-quantitatively measured using the Gel Doc system with a CCD camera and then statistically analyzed in order to determine which component of TJ-9 was the true cytokine inducer. The results showed that the scutellaria root is the main component inducing the cytokines, while the glycyrrhiza root is the secondary component. When the cytokine concentrations in the supernatants of cell cultures were measured by ELISA, the levels of IL-12, IL-1beta, IL-10, TNF-alpha, and G-CSF reflected mRNA expression levels in the cell fraction. However, the level of IFN-gamma was below the detectable limit. The effects of various reagents on many different kinds of cytokine mRNA expression could be analyzed objectively in a short time using the Gel Doc system. Many important findings could be demonstrated by this simple, easy, sensitive, and cheap method. After the clinical significance of cytokine analysis is confirmed, this method may become a useful clinical examination tool. PMID- 11436204 TI - Comparative study of cardiac troponin I and T measurements in a routine extra cardiological clinical setting. AB - This study compared troponin I (cTnI) to troponin T (cTnT) in a population admitted to General Medicine Divisions in whom acute myocardial infarction (AMI) was suspected; 98 consecutive patients were included. Diagnoses were made without knowledge of troponin results: 51 patients had AMI, and 47 (including 8 with unstable angina) had no AMI. Patients were considered to be troponin positive if the marker concentration was >99th percentile value of the reference population. Both troponins were associated with an almost absolute sensitivity for AMI (100% for cTnI and 98.0% for cTnT), while the specificity was marginally higher for cTnI (78.7% vs. 68.1%). Increased cTnI and/or cTnT were observed in 15 patients out of 39 without acute coronary syndromes. Simultaneous positivity was seen in 8 patients with severe disorders and complications. Discordances were more frequent in favor of increased cTnT (n = 5) than the opposite (n = 2), even if this difference did not achieve statistical significance. cTnI and cTnT detected AMI with comparable efficiency. Cases without coronary syndrome positively concordant for troponins confirmed the ability of these biomarkers to detect myocardial injury undetectable by conventional diagnostic approaches. PMID- 11436205 TI - Quantitation of TGF-beta1 mRNA in porcine mesangial cells by comparative kinetic RT/PCR: comparison with ribonuclease protection assay and in situ hybridization. AB - Gene expression can be examined with different techniques including ribonuclease protection assay (RPA), in situ hybridisation (ISH), and quantitative reverse transcription-polymerase chain reaction (RT/PCR). These methods differ considerably in their sensitivity and precision in detecting and quantifying low abundance mRNA. Although there is evidence that RT/PCR can be performed in a quantitative manner, the quantitative capacity of this method is generally underestimated. To demonstrate that the comparative kinetic RT/PCR strategy-which uses a housekeeping gene as internal standard-is a quantitative method to detect significant differences in mRNA levels between different samples, the inhibitory effect of heparin on phorbol 12-myristate 13-acetate (PMA)-induced-TGF-beta1 mRNA expression was evaluated by RT/PCR and RPA, the standard method of mRNA quantification, and the results were compared. The reproducibility of RT/PCR amplification was calculated by comparing the quantity of G3PDH and TGF-beta1 PCR products, generated during the exponential phases, estimated from two different RT/PCR (G3PDH, r = 0.968, P = 0.0000; TGF-beta1, r = 0.966, P = 0.0000). The quantitative capacity of comparative kinetic RT/PCR was demonstrated by comparing the results obtained from RPA and RT/PCR using linear regression analysis. Starting from the same RNA extraction, but using only 1% of the RNA for the RT/PCR compared to RPA, significant correlation was observed (r = 0.984, P = 0.0004). Moreover the morphometric analysis of ISH signal was applied for the semi-quantitative evaluation of the expression and localisation of TGF-beta1 mRNA in the entire cell population. Our results demonstrate the close similarity of the RT/PCR and RPA methods in giving quantitative information on mRNA expression and indicate the possibility to adopt the comparative kinetic RT/PCR as reliable quantitative method of mRNA analysis. PMID- 11436206 TI - Direct measurement of HDL cholesterol: method eliminating apolipoprotein E-rich particles. AB - It has been reported that the existing direct method of high density lipoprotein (HDL) cholesterol measures particles enriched with apolipoprotein E (apoE). The aim of our study was to investigate a new analytical protocol to directly measure HDL cholesterol that eliminates apoE-rich particles. The interactions of four lipoproteins (HDL(3), HDL(2), LDL, and VLDL + chylomicron) with surfactants, divalent cations, sugars, and lectins were investigated. By analyzing sera, HDL(3), and HDL(2), we examined the relationships among the measurements obtained by our protocol, a precipitation method using heparin-MnCl(2), and a commercially available kit for this direct method. A significant difference was found between the direct method and the heparin-MnCl(2) method, but not between our protocol and the heparin-MnCl(2) method. Multiple regression analysis showed that the difference between the direct method and the heparin MnCl(2) method is dependent on sources of apoE-rich HDL. In conclusion, our protocol enables a direct measurement of HDL cholesterol that eliminates apoE-rich particles. PMID- 11436207 TI - An efficient chemical shift imaging scheme for magnetic resonance-guided neurosurgery. AB - An efficient magnetic resonance spectroscopic imaging (MRSI) or chemical shift imaging (CSI) technique based on multiple spin echoes (MSE) has been implemented, validated, and used in both phantom and in vivo MR-guided neurosurgical applications. The key concept of the method is to employ MSE to significantly speed up the data collection rate for mapping hydrogen-containing metabolites. Using an echo train length (ETL) of three per excitation to simultaneously fill three consecutive k-space areas, the total scan time for a spectroscopic image matrix size of 32 x 32 has been shortened to approximately 11 minutes. An interecho spacing time of 273 msec was used to null the phase anomalies of lactate double peaks due to the J-coupling. This allowed a sufficient long data sampling time to achieve 4 Hz spectral resolution. Performing CSI intraopertively during an MR-guided neurosurgical procedure was shown to be feasible at 1.5 T. More importantly, it was shown that more relevant information can be obtained regarding neurochemistry about a targeted lesion, in addition to conventional MR morphological imaging noninvasively. In 25 MR-guided neurosurgical cases, the alleviated choline signal has been found to be consistent with the existence of rapid tumor cell proliferation in the corresponding area. The actual neurobiopsy guided by the spectroscopic imaging method demonstrated that it could provide valuable information in specifying the optimal site in a biopsy procedure, especially in the case involving a nonenhancing tumor. The multiecho scheme has made the CSI technique efficient enough to be routinely used in MR-guided surgical procedures at 1.5 T and also allows the possibility of taking full advantage of MRI capability. PMID- 11436208 TI - Spectroscopic increase in choline signal is a nonspecific marker for differentiation of infective/inflammatory from neoplastic lesions of the brain. AB - We report in vivo proton magnetic resonance (MR) spectroscopic findings in three benign infective/inflammatory lesions (one case each of tuberculoma, fungal granuloma, and xanthogranuloma), which showed high choline along with the presence of lipid/lactate, a feature characteristically described in neoplastic lesions. Histopathology of the lesions showed inflammatory cellular infiltrates with areas of necrosis/caseation. The spectroscopic-visible increased choline resonance in these lesions is probably the result of cellularity. We conclude that increased choline, along with the presence of lipid/lactate is a nonspecific finding and may not be of much value in the differentiation of neoplastic from nonneoplastic infective/inflammatory intracranial mass lesions. PMID- 11436209 TI - The effects of ACE inhibitor therapy on left ventricular myocardial mass and diastolic filling in previously untreated hypertensive patients: a cine MRI study. AB - Cardiac remodeling in case of hypertension induces hypertrophy of myocytes and elevated collagen content and, subsequently, impaired diastolic filling of the left ventricle. The purpose of this prospective study was to evaluate changes of left ventricular (LV) myocardial mass, as well as diastolic filling properties, in hypertensive patients treated with the ACE inhibitor fosinopril. Sixteen hypertensive patients with echocardiographically documented LV hypertrophy and diastolic dysfunction received fosinopril (10-20 mg daily). Measurements of LV myocardial mass and properties of diastolic filling (peak filling fraction (PFF); peak filling rate (PFR)) were performed prior to medication, as well as after 3 and 6 months of therapy using cine magnetic resonance imaging (MRI). Ten healthy subjects served as a control group. LV myocardial mass (g/m2) decreased continuously within 3-6 months of follow-up by 32% (148 +/- 40 vs. 120 +/- 26 vs. 101 +/- 22 g/m2; P < 0.0001/0.005). The extent of regression correlated to the severity of LV hypertrophy at baseline (r = 0.77; P < 0.004). Early diastolic filling increased significantly within 6 months of therapy (PFF (%): 36 +/- 6 vs. 61 +/- 7, P < 0.0001; PFR (mL/second): 211 +/- 48 vs. 282 +/- 48, P < 0.001). Cine MRI can be used to assess the time course of pharmacological effects on cardiac remodeling in the course of hypertension. ACE inhibitor therapy results in a significant reduction of LV mass within 3 months and is accompanied by a normalization of diastolic filling that is completed after 6 months. PMID- 11436210 TI - Qualitative and quantitative analysis of regional left ventricular wall dynamics using real-time magnetic resonance imaging: comparison with conventional breath hold gradient echo acquisition in volunteers and patients. AB - A real-time magnetic resonance imaging (MRI) acquisition sequence was evaluated for the assessment of left ventricular wall motion (WM) and wall thickening (WT). Ten normal volunteers and 21 patients were studied. Short-axis cine images of the left ventricle (LV) were acquired with a fast gradient echo and an ultrafast segmented echo-planar imaging (EPI) sequence. Qualitative and quantitative analysis of WM and WT was performed on a segmental basis. Qualitative scores agreed between the two methods in 691 of 724 segments (95.4%) with good reproducibility. Quantitative measurements of WM and WT were significantly lower (P < 0.001) with the real-time method (WM: mean bias, 0.49 mm; WT: mean bias, 0.61 mm). The largest differences were observed in the anterior and lateral segments and in patients with dilated ventricles. The lower resolution of the real-time sequence and artifacts was probably responsible for these differences. In conclusion, real-time cardiac MRI can be used for qualitative assessment of wall dynamics but is presently insufficient for quantitative analysis. PMID- 11436211 TI - Magnetic resonance planimetry of the vena contracta as a new approach to assessment of stenotic heart valves: an in vitro study. AB - Exact determination of the orifice area in stenotic valve disease is essential to guide therapy. With the standard imaging methods, the orifice area has to be calculated by empirically-derived formulas, which may be susceptible to changes in hemodynamic status, leading to wrong therapeutic decisions. We tested the ability of magnetic resonance imaging (MRI) to quantify the orifice area by planimetry of the proximal vena contracta (PVC) in an idealized, constant-flow model in a 1.0-T tomograph (Siemens Magnetom Expert). There was a close correlation of the PVC area in MRI and the true orifice area (r2 = 0.985), and agreement of the methods as measured by the Bland-Altman test was significant with a low systematic error of -0.02 cm2. We conclude that MRI planimetry of the PVC in stenotic valves in a constant-flow model may be used to reliably measure the orifice area. PMID- 11436212 TI - In vivo magnetic resonance (MR) study of fatty liver: importance of intracellular ultrastructural alteration for MR tissue parameters change. AB - Fatty liver is thought to have a shorter T1 relaxation time than normal liver tissue, due to the accumulation of triglyceride. Previous studies regarding T1 and T2 relaxation times, however, show widely different results. In order to elucidate the mechanism responsible for the changes and diversity of relaxation times in fatty liver, we created two animal models in 14 rabbits, one acute form (N = 6) and the other chronic form (N = 8). Four rabbits were taken as a control group. Tissue relaxation times and the magnetization transfer (MT) effect of the liver tissue in these two models were measured. The results were correlated with biochemical analysis of water and fat content and histological examination, including findings in light microscopy and electron microscopy. Although the fatty ratio in both forms of fatty liver was similar, their tissue relaxation rate and MT effect were significantly different. The acute form showed prolongation of both T1 and T2 relaxation times (512 +/- 51 msec vs. 710 +/- 95 msec and 39 +/- 1.8 msec vs. 48 +/- 3.7 msec, respectively) and a decrease of the MT effect (50 +/- 5.1% vs. 38 +/- 6.3%), compared to those of the control group and preinduction liver. The chronic form showed shorter T1 and T2 values (526 +/- 36 msec vs. 406 +/- 56 msec and 36 +/- 1.6 msec vs. 33 +/- 2.3 msec, respectively) and a stronger MT effect (21 +/- 0.9% vs. 26 +/- 2.3%). In acute form fatty liver, electron microscopic examination revealed dramatic subcellular changes, such as vesicular transformation, a markedly increased amount of smooth endoplasmic reticulum (SER), and disruption of the crista. These changes were not found in the chronic form fatty liver. From this study, we concluded that the ultrastructural alteration in the subcellular organelles of hepatocyte might play a crucial role for the chameleonic presentation of MR tissue parameters in fatty liver. PMID- 11436213 TI - Diffusion tensor MRI of the human kidney. AB - This study characterizes the diffusion anisotropy of the human kidney using a diffusion-weighted, single-shot echo planar imaging (EPI) sequence in order to access the full apparent diffusion tensor (ADT) within one breathhold. The fractional anisotropy (FA) of the cortex and the medulla were found to be 0.22 +/ 0.12 and 0.39 +/- 0.11, respectively (N = 10), which emphasizes the need for rotationally invariant diffusion measurements for clinical applications. Additional limitations for clinical diffusion imaging on the kidney are the strong susceptibility variations within the abdomen that restrict the use of imaging techniques employing long echo trains, and the severe motion sensitivity that limits the available imaging time to one breath-hold. To overcome these problems an isotropic, diffusion-weighted, segmented EPI protocol that facilitates the acquisition of high-resolution diffusion-weighted images within a single breath-hold was implemented. Using this method, the apparent diffusion coefficient (ADC) of the cortex and medulla were found to be 2.89 +/- 0.28. 10( 9) m2/s and 2.18 +/- 0.36. 10(-9) m2/s (N = 10). PMID- 11436214 TI - Spatial variation in cartilage T2 of the knee. AB - Technical limitations imposed by resolution and B1 homogeneity have thus far limited quantitative in vivo T2 mapping of cartilage to the patella. The purpose of this study is to develop T2 mapping of the femoral/tibial joint and assess regional variability of cartilage T2 in the knee. Quantitative in vivo T2 mapping of the knee was performed on 15 asymptomatic adults (age, 22-44) using a 3T MR scanner. There is a consistent pattern of spatial variation in cartilage T2 with longer values near the articular surface. The greatest variation occurs in the patella, where T2 increases from 45.3 +/- 2.5 msec at a normalized distance of 0.33-67 +/- 5.5 msec at a distance of 1.0. These results demonstrate feasibility of performing in vivo T2 mapping of femoral tibial cartilage. Except for the superficial 15% where T2 values are lower, the spatial variation in T2 of femoral and tibial cartilage is similar to patellar cartilage. PMID- 11436215 TI - A multielement RF coil for MRI guidance of interventional devices. AB - Accurate localization of minimally invasive devices is critical to the success of interventional procedures. Device orientation and tip position are two of the most important pieces of information needed to define device location for magnetic resonance imaging (MRI)-guided interventional procedures. While a single one-element micro coil incorporated into an interventional device has proven to be effective in some applications, it can only supply tip position information. However, multiple positions on the device are necessary to also determine its orientation. For this purpose, a novel single micro coil design with three separate winding elements that provides both the device orientation and tip position is described in this study. Definition of MR scan planes, by using the device orientation and the target tissue location, permits automatic tracking of the insertion of the device. Furthermore, devices that include this coil design are permitted to bend to a limited extent. This makes the micro coil design appropriate for many flexible interventional devices. Reliable near-real-time tracking of three points on an interventional device is demonstrated on a 0.2T MRI system with modest gradient performance. Phantom and in vivo animal experiments are used to demonstrate the utility of this new coil design. PMID- 11436216 TI - Three-dimensional adaptive filtering in magnetic resonance angiography. AB - In order to enhance 3D image data from magnetic resonance angiography (MRA), a novel method based on the theory of multidimensional adaptive filtering has been developed. The purpose of the technique is to suppress image noise while enhancing important structures. The method is based on local structure estimation using six 3D orientation selective filters, followed by an adaptive filtering step controlled by the local structure information. The complete filtering procedure requires approximately 3 minutes of computational time on a standard workstation for a 256 x 256 x 64 data set. The method has been evaluated using a mathematical vessel model and in vivo MRA data (both phase contrast and time of flight (TOF)). 3D adaptive filtering results in a better delineation of small blood vessels and efficiently reduces the high-frequency noise. Depending on the data acquisition and the original data type, contrast-to-noise ratio (CNR) improvements of up to 179% (8.9 dB) were observed. 3D adaptive filtering may provide an alternative to prolonging the scan time or using contrast agents in MRA when the CNR is low. PMID- 11436217 TI - Relaxation effects of clustered particles. AB - Relations between spatial distribution of superparamagnetic iron oxide (SPIO) particles and the image contrast caused by SPIO were investigated. Actual clustering pattern of particles was measured in the liver and spleen of animals using intravital laser confocal microscopy. SPIO-doped phantoms with and without Sephadex beads were made to simulate these patterns, and relaxation parameters were measured using a 1.5-T clinical scanner. Finally, these results were compared to clinical image data using SPIO particulate agent. Intravital microscopy indicated that the clustering of latex beads was more predominant in hepatic Kupffer cells than in splenic macrophages (P < 0.001). Phantoms without Sephadex beads showed an approximately linear increase of 1/T1 (R1), 1/T2 (R2) and 1/T2* (R2*) values with increasing SPIO concentration. However, with Sephadex beads, R1 and R2 showed little change with increasing SPIO concentration, while R2* showed the same linear increase with SPIO. Also, the R2* values were higher with Sephadex beads. These results were consistent with the clinical imaging data, where signal reduction was significantly smaller in the spleen (-0.4% +/- 27.4%) than in the liver (50.4% +/- 16.8%, P < 0.00001) on T2*-weighted images, but the reduction in the spleen (47.2% +/- 16.1%) was equivalent to the liver (38.8% +/- 26.0%) on T2-weighted images. PMID- 11436218 TI - Proton magnetic resonance spectroscopy of choroid plexus tumors in children. AB - A variety of lesions may present as intraventricular masses in children. We report quantitative proton magnetic resonance spectroscopy (MRS) of two intraventricular tumors of the choroid plexus: choroid plexus carcinoma (CPC) and choroid plexus papilloma (CPP). Both lesions were characterized by high levels of choline-containing compounds and a complete absence of creatine and the neuronal/axonal marker N-acetyl aspartate. The CPC showed higher levels of choline compared to the CPP, and it also had elevated lactate. These preliminary results, if confirmed in a larger cohort of patients, indicate that proton MRS may have a role in the presurgical diagnosis of choroid plexus tumors in children, which may also have important implications for therapy and prognosis. PMID- 11436219 TI - Magnetic resonance microscopy at 17.6-Tesla on chicken embryos in vitro. AB - The non-destructive nature and the rapid acquisition of a three-dimensional image makes magnetic resonance microscopy (MRM) very attractive and suitable for functional imaging investigations. We explored the use of an ultra high magnetic field for MRM to increase image quality per image acquisition time. Improved image quality was characterized by a better signal-to-noise ratio (SNR), better image contrast, and higher resolution compared to images obtained at lower magnetic field strengths. Fixed chicken embryos at several stages of development were imaged at 7.0-T (300 MHz) and at 17.6-T (750 MHz). Maximum intensity projection resulted in three-dimensional vascular images with ample detail of the embryonic vasculature. We showed that at 750 MHz frequency, an image with approximately three times better SNR can be obtained by T1-weighting using a standard gadolinium contrast agent, compared to the same measurement at 300 MHz. The image contrast improved by around 20 percent and the contrast-to-noise ratio improved by almost a factor of 3.5. Smaller blood vessels of the vascular system were identified at the high field, which indicates a better image resolution. Thus, ultra high field is beneficial for MRM and opens new areas for functional imaging research, in particular when SNR, resolution, and contrast are limited by acquisition time. PMID- 11436220 TI - Autocorrelation analysis of bone structure. AB - We propose a method called spatial autocorrelation analysis (SACA) to determine the spatial anisotropy of the trabecular bone in order to investigate osteoporosis. For demonstrating the potential of SACA we first evaluate the method on rectangular, simulated test patterns as a simple model for the anisotropic pore structure of the bone. As a next step towards biomedical application, photographic reference images of human vertebral bone were investigated by SACA. Osteoporotic bone structure could be clearly differentiated from non-osteoporotic sample images. Moreover, for demonstration of the applicability and potential of the method for in vivo characterization of osteoporosis, the microstructure of the human calcaneus was investigated by MR microimaging on a young healthy male subject and an osteoporotic female. The measurements were performed using a high-field (3T) whole-body MR tomograph equipped with a special, strong head gradient system. The signal was acquired with a surface coil mounted on an in-house-built device for convenient immobilization of the subject's foot. Using a 3D gradient echo sequence a resolution of 0.254 x 0.254 x 2.188 mm3 was achieved in vivo. Selected images were inverted, gradient corrected for the inhomogeneous but sensitive detection by the surface coil, and subsequently analyzed by SACA. The anisotropy of bone structure detected by SACA is a possible candidate for noninvasive determination of the osteoporotic status, potentially complementing standard bone mineral density measurements. PMID- 11436221 TI - Water relaxation by SPM particles: neglecting the magnetic anisotropy? A caveat. AB - Magnetometric and relaxometric data from SPM particles are revisited, leading to a double conclusion rather different from the original one: accounting for the anisotropy energy not only allows a considerable improvement of the fit of the data to theory, but it also invalidates the conclusion obtained from the high field data (without accounting for anisotropy) concerning an important reduction of the diffusion coefficient by the coating of the particles. PMID- 11436222 TI - Index of suspicion. Case 2. Diagnosis: Norrie disease. PMID- 11436224 TI - Retraction: A SADS defect in tumor cells provides optimism. PMID- 11436223 TI - Index of suspicion. Case 3. Diagnosis: Infantile pseudotumor cerebri. PMID- 11436225 TI - Disbribution of head and neck cancer in UK. PMID- 11436227 TI - The dental examination: the bare essentials. PMID- 11436226 TI - Research funds for complementary medicine. PMID- 11436228 TI - The new Oral Hygienists Association of South Africa. PMID- 11436229 TI - An open reply to a community dentist. PMID- 11436230 TI - Continuous payment problems: MX-health. PMID- 11436231 TI - Hyperresponders and adrenaline in local anaesthetic solutions. AB - The presence of adrenaline in local anaesthetic solutions may cause disturbing systemic reactions in some patients. Putative allergies of 532 patients to local anaesthetics were investigated over a 15-year period in a clinic and 163 patients were found to experience these reactions, of whom 99% were female. Only 17 of these patients took drugs for the treatment of cardiovascular diseases; 11 of them were on thyroid replacement therapy. Practitioners should be careful not to attribute these systemic reactions to an allergic drug response. Furthermore, the medical history and drug intake of patients must be revised regularly. PMID- 11436232 TI - Anti-plaque efficacy of a chalk-based antimicrobial dentifrice. AB - A chalk-based antimicrobial dentifrice was developed and has undergone in vitro testing to determine antimicrobial efficacy by kill-time studies, and in vivo clinical studies to determine anti-plaque build-up properties. The antimicrobial efficacy of the formulation is attributable to the following components; sodium lauryl sulphate, sodium monofluorophosphate, flavouring oils, triclosan, and low water activity. In vitro kill-time testing showed the dentifrice to have good activity against organisms associated with plaque, showing at least 99% reductions in the numbers of each organism. In vivo clinical results showed the dentifrice to protect against the build-up of plaque when compared to water alone. These results demonstrate that this dentifrice is effective in controlling plaque, and can thus be expected to promote good oral hygiene. PMID- 11436233 TI - Examiner performance with visual, probing and FOTI caries diagnosis in the primary dentition. AB - To compare clinical reproducibility of dental caries diagnosis in the primary dentition under field conditions, a convenience sample of 5-year-old children in a nursery school in Germiston, was examined for dental caries by four dentists using visual (mirror), visual plus tactile (mirror plus probe) and fibre-optic transillumination (FOTI) methods. Seventeen children were examined on day one and 11 re-examined on day two. Inter-examiner agreement was high, above 90%. Visual examination on its own is comparable with the traditional visual plus tactile method and to FOTI under field conditions. New caries data collected by visual diagnosis alone may, reasonably, be compared with historical data diagnosed with visual + tactile examination. PMID- 11436234 TI - Dentine caries: take it or leave it? AB - In modern dentistry the primary aim when excavating carious dentine is to eradicate only the highly infected, irreversibly demineralised and denatured biomass in order to allow effective restoration of the cavity, restoration of the surface anatomy of the tooth and to prevent disease progression. However, the boundary between this superficial zone of dentine requiring excavation and the deeper, affected but repairable tissue is not always obvious either in the clinic or in the research laboratory. The inherent subjectivity in detecting this excavation boundary can result in clinically significant differences in the quality and quantity of dentine removed by different operators and makes the in vitro comparison of newer excavation techniques more difficult. This article discusses the rationale behind carious dentine excavation and the criteria available to the dentist, both clinical and laboratory, to help identify the dentine requiring removal. PMID- 11436235 TI - Curriculum 2001: a proposal for a new, integrated, dental curriculum at the University of the Witwatersrand. AB - Recent technological advances and an escalation of knowledge in dentistry have highlighted the need to revise the current undergraduate dental curriculum at the University of the Witwatersrand. This imperative is reinforced on one hand by manifest problems in content, structure and sequencing of the existing curriculum, and on the other hand by the need to urgently address contemporary South African issues. Revision will also offer the opportunity to incorporate modern educational practice and philosophy. This paper proposes that the contents, sequence and duration of the curriculum must be determined by the knowledge demands of dentistry (global issues) as well as by local constraints. It suggests early clinical contact and changes in syllabus content, pedagogy and assessment practices in an undergraduate course of slightly reduced duration. PMID- 11436236 TI - An 8-year assessment of maxillofacial prosthetic patients treated in a Department of Prosthetic Dentistry. AB - The Department of Prosthetic Dentistry at the University of the Witwatersrand, Johannesburg, serves as a referral centre for maxillofacial prosthetic patients from the surrounding general hospitals and local private clinics. This places a time and financial constraint on the department. The aim of this study was to establish the number of new patients seen annually, the causative factors associated with the various types of defects, the treatment provided and the cost per patient in terms of clinical hours and prosthesis fees. The data were taken from the records of patients treated in the maxillofacial prosthetics clinic during an 8-year period, from January 1991 to December 1998. The total number of new patients was calculated and the data were collated according to the year in which treatment commenced. Results were tabulated according to the number of new cases in each group, the aetiology of the defects, prosthetic treatment provided and the costs in terms of clinical hours and prostheses. The results of this study indicate that there is a need for a specialised Maxillofacial Prosthetic Unit. Although the present fee schedule needs to be revised, there is still a need for a substantial amount of state subsidy. PMID- 11436237 TI - Patient management: measuring patients' expectations and perceptions of service quality in a dental training hospital. AB - The difference between service quality expectations and perceptions (experiences) of patients (customers) attending a dental training hospital was investigated by using a modified version of the Parasuraman SERVQUAL model. A questionnaire comprising 28 service quality-related statements and four open-ended questions was used at the interviews. The study showed that 11.6% of respondents experienced problems with the service. A principal component factor analysis indicated that two of the five dimensions of service quality, namely reliability and assurance, contributed to 59% of service level variance. Female patients showed larger mean differences than male patients. The greater the number of visits to the hospital, the smaller the difference between expectations and perceptions. Patients in the category 36-45 years of age, showed larger mean differences than younger or older patients. Respondents with no academic qualifications had lower expectations of the service, while professional people seemed to have more realistic expectations prior to a visit to the hospital than respondents in the technical/clerical category. PMID- 11436238 TI - Rwanda to receive cheaper anti-HIV drugs for chronic treatment and free viramune to prevent viral transmission from mother to child. PMID- 11436240 TI - Why integrated health networks have failed. AB - Over the last two decades, hospitals, physicians, and nursing homes have rushed to merge or partner with one another, cheered on by consultants, academics, and experts who claimed that such networking was imperative for these organizations to survive in an increasingly competitive environment. In the 1990s, however, reports of the major difficulties encountered by the merged entities began to dampen the industry's enthusiasm for integration, causing some to even question the value of the integration "revolution." This article will review the primary rationale behind integration and will assess its success to date. Based on field reports and the available evidence, the authors conclude that IHNS have not delivered on their promise. While each network should be considered in its unique market and contextual situation, the enormous financial, human, and clinical resources devoted to integration have not borne much fruit. Evidence of quantifiable, sustained financial or clinical value is scant. PMID- 11436241 TI - Integrated healthcare is alive and well. PMID- 11436242 TI - Integrated health networks are not created equal. PMID- 11436243 TI - Have IHNS failed in healthcare? PMID- 11436244 TI - Mold litigation--serious business. PMID- 11436245 TI - Psychoanalysis and cognitive behaviour therapy--rival paradigms or common ground? AB - The author suggests that contemporary enthusiasm for cognitive-behavioural therapy reflects our longing for swift, rational help for psychological suffering. Competition for funding threatens the psychoanalytic presence in the public sector. The psychoanalytic and cognitive-behavioural models are contrasted, and the relative richness of the psychoanalytic paradigm outlined. The author suggests that a cognitive model is commonsensical, but less complex, with less potential explanatory and therapeutic power. She discusses how the analytic stance is always under pressure to 'collapse' into simpler modes, one of which resembles a cognitive one. This also occurs inevitably, she argues, when attempts are made to 'integrate' the two models. Cognitive and 'integrated' treatments nevertheless have the advantage that they are less intrusive and hence more acceptable to some patients. Selected empirical process and outcome research on cognitive and psychoanalytic therapies is discussed. Brief psychotherapies of either variety have a similar, modestly good outcome, and there is some evidence that this may be based more on 'dynamic' than 'cognitive' elements of treatment. Formal outcome studies of more typical psychoanalytic psychotherapy and of psychoanalysis itself begin to suggest that these long and complex treatments are effective in the more comprehensive ways predicted by the model. PMID- 11436246 TI - Revising psychoanalytic interpretations of the past. An examination of declarative and non-declarative memory processes. AB - The author reviews a contemporary cognitive psychology perspective on memory that views memory as being composed of multiple separate systems. Most researchers draw a fundamental distinction between declarative/explicit and non declarative/implicit forms of memory. Declarative memory is responsible for the conscious recollection of facts and events--what is typically meant by the everyday and the common psychoanalytic use of the word 'memory'. Non-declarative forms of memory, in contrast, are specialised processes that influence experience and behaviour without representing the past in terms of any consciously accessible content. They operate outside of an individual's awareness, but are not repressed or otherwise dynamically unconscious. Using this theoretical framework, the question of how childhood relationship experiences are carried forward from the past to influence the present is examined. It is argued that incorporating a conceptualisation of non-declarative memory processing into psychoanalytic theory is essential. Non-declarative memory processes are capable of forming complex and sophisticated representations of the interpersonal world. These non-declarative memory processes exert a major impact on interpersonal experience and behaviour that needs to be analysed on its own terms and not mistakenly viewed as a form of resistance. PMID- 11436247 TI - Incentives for a reconsideration of the debate on metapsychology. AB - With a view to establishing an area of metapsychological common ground, the author examines the attempts made by G. S. Klein, Gill and Brenner to interpret Freud's conception of metapsychology. Quoting liberally from the correspondence with Fliess and from works composed at all stages of Freud's career, the author shows that the versions put forward by Klein and Gill do not stand up to scrutiny; the economic aspect on which Freud himself insisted and which Brenner upholds is also found wanting. It is argued that, if the Freudian 'co-ordinates' of metapsychology are to have an epistemological function, they must have a solid foundation in the representational world to which the psychoanalytic process affords access. Whereas this is the case with the topographical, dynamic and structural viewpoints, it is not true of the economic aspect. Energies are stated to exist in the representational world only in the form of affects, so that the economic aspect should, in the author's opinion, be abandoned in favour of an affective one. In the context of the endeavour to obtain pleasure and avoid unpleasure adduced by Freud, this viewpoint would concentrate on the relations between affects and the other key elements of the representational world to which the other metapsychological parameters relate. PMID- 11436248 TI - Vicissitudes of bisexuality. Crucial points and clinical implications. AB - The author begins by noting that the heuristic value of the concept of bisexuality can be explored in two directions, involving respectively its role in psychopathology and in the development of creativity and thought. The author concentrates on the first of these aspects, which entails a reconsideration of the 'phallic question'. The status of femininity is investigated on the basis of Freud's and Winnicott's theories of bisexuality and the author here finds an explanation for the repudiation of femininity and for the 'phallic question' thereby implied, which represents the underlying bedrock resulting in interminable analyses. An important element in this rejection is stated to be the maternal aspect of femininity. Particular reference is made to the somatic roots of phallic defences and to their effects on thought and symbolisation. Some brief clinical examples are presented to suggest that the bedrock may be due to the distorted use of sensoriality and of the erotogenic zones. The author contends that the anal zone plays a crucial part in the manifestations of bisexuality and contrasts phallic logic with the logic of triangulation. Numerous literature references and clinical examples support and illustrate her arguments throughout. The author ends with a discussion of the implications of bisexuality for thought and knowledge. PMID- 11436249 TI - The drive circuit as generator of subjectivation. AB - It is a fact that Freud almost systematically refrained from using the term 'subject' in his work. However, one remarkable exception is worthy of our attention: in 'Instincts and their vicissitudes' (1915), he repeatedly refers to the notion of subject (Subjekt) whenever he is dealing with one common 'vicissitude' (destiny) of basic drive pairs, the one in which there is a succession of the 'turning round upon one's own body' and the 'reversal of the aim into its opposite', i.e. the search for satisfaction in a passive mode (to get looked at, taken in hand etc.); here Freud designates the indispensable exterior agent of the active drive as a 'new subject'. The author finds in this the beginnings of a specifically psychoanalytic conception of the subject function, the latter gaining its substance from being agent of the drive interplay itself, with its unconscious dimension, and thus distinguishing itself from the functions reserved to the ego. He suggests that the originating matrix of the process of subjectivation be located between the infant's instinctual reaching out and the parental partner's response, which will take on more and more signifying value. The treatment of a bulimic patient is presented to illustrate the problems of a process of subjectivation that must develop by overcoming the trap of oral servitude. PMID- 11436250 TI - Collaborating with the unconscious other. The analysand's capacity for creative thinking. AB - The analysand's capacity for making use of psychoanalytic treatment has been a subject of importance since the beginning of psychoanalysis. The author addresses an aspect of the difficulty encountered by analysands in achieving a psychic state that allows the creative use of free association, dreams, parapraxes and other spontaneous phenomena occurring during the course of treatment. He suggests that a very specific state of mind is essential to both the psychoanalytic process and the creative process. Using theoretical concepts derived from Freud, Klein and Bion, he develops the idea of an internal object relationship, 'the collaboration with the unconscious other', which forms the basis for both creative thinking and the psychoanalytic function of the personality. Creative thinking is distinguished from artistic endeavour and discussed as a universal potential, on which growth in psychoanalysis depends. The term 'unconscious other' is meant to signify the subjective experience of a foreign presence within oneself from which both spontaneous creative inspiration and involuntary psychic phenomena are felt to emanate. The author presents clinical material to suggest that paranoid-schizoid and depressive anxieties form obstacles to collaborating with the unconscious other, and must be worked through in order to achieve an analytic process. PMID- 11436251 TI - Mapping the landscape: levels of transference interpretation. AB - The author argues that there are distinctly different kinds of transference interpretation, each of which might be valid in particular circumstances in analysis, but which contain and imply different understandings of what is meant by a 'transference interpretation'. She suggests that transference interpretations may be at any one of four different levels, and she describes these levels as ranging from interpretations that point to links between current events in the analysis and events from the patient's history, through interpretations that link events in the patient's external life to the patient's often unconscious phantasies about the analyst and the analysis, to interpretations that focus on the use of the analyst and the analytic situation to enact unconscious phantasy configurations, sometimes pulling the analyst into the enactment. Material from four consecutive sessions of an analysis is presented to illustrate how all levels of transference interpretation may be part of a lively and meaningful analysis, but how the level of interpretation may change as the level of understanding deepens within a session and from one session to the next. PMID- 11436252 TI - Whose sleep is it, anyway? Or 'night moves'. AB - The analyst's 'sleep' during sessions is a puzzling, troubling, extreme experience, which has rarely been described in the psychoanalytic literature. The author presents a clinical illustration in which her recurring 'sleep' during the sessions was approached as an open, central issue. She attempts to explore, understand and integrate this experience theoretically and clinically, first by reviewing and examining the psychoanalytic literature on the subject and on related phenomena, and then, more particularly, by formulating her own explanation of it. She emphasises being in the grip of the psychoanalytic process, and the immersed involvement and converging of patient and analyst, which generate a conjoint state of deep experiential interconnectedness and impact on each other--in particular the impact of the patient's inner world on the analyst. In this context, the author also refers to the notions of 'the uncanny', 'fear of breakdown' and dissociative self-states and the mitigation of the patient's dissociative self-experience via the analyst's vicarious dissociative experience. PMID- 11436253 TI - The psychotherapy of a male anorectic. AB - The author describes the way in which a male anorexic patient came to be understood in once-weekly psychotherapy. The findings are similar to recent psychoanalytic ones about female anorectics, but add that he was preoccupied with display and that there was an erotised transference in the oral mode. Of two major themes, the first was a food obsession; the food was felt concretely to be the mother as in a symbolic equation, and the conflict about eating represented a wish for and fear of fusion with her. The second major theme concerned exhibition; the self, as seen in two screen memories, was felt to be uncomfortably on show. Meanwhile the patient was preoccupied with a self displaying maternal object, which was prominent in the transference. This perception of the object served as a defence against envy, although the defensive aspect may have been added to by an actual experience of a mother who functioned operatively, giving food rather than loving attention. The patient employed primary-process thinking defensively against self-object differentiation, which would have excited further awareness both of envy and of the real deficiencies of his primary objects. The author raises the possibility that the anorexia functioned as a perversion, binding and organising the core complex about fusion. PMID- 11436255 TI - The place of affect in the representational world: in memory of Joseph Sandler. PMID- 11436254 TI - The unconscious and primal phantasies. PMID- 11436256 TI - On Lawrence's 'Loving them to death: the anorexic and her objects'. PMID- 11436257 TI - Marion Milner 1900-1998. PMID- 11436258 TI - The facts about vCJD. PMID- 11436259 TI - A raised PSA from a private medical. PMID- 11436260 TI - Transient ischaemic attacks: a GP guide. PMID- 11436261 TI - Understanding cerebral tumours. PMID- 11436262 TI - Managing the patient with migraine. PMID- 11436263 TI - The treatment of motor neurone disease. PMID- 11436264 TI - The mouth in neurological disorders. PMID- 11436265 TI - Advances in therapeutic neurology. PMID- 11436266 TI - Reducing renal disease in diabetes. PMID- 11436267 TI - The use of syringe drivers in the community. PMID- 11436268 TI - A spot of Haslam's disease. PMID- 11436269 TI - Towards the control of a powered orthosis for people with muscular dystrophy. AB - The paper describes the development of a passive/active orthosis for people with limited anti-gravity strength in their arms. This is symptomatic of conditions such as muscular dystrophy and spinal muscular atrophy. A passive orthosis was designed and developed using linear elastic elements. The system is being tested with children with disabilities and preliminary results are encouraging. An RT200 robot was also used as a test-bed for an active orthosis. The robot was instrumented with a six-axis force/torque sensor at the end-effector. The force acted as the input to the robot. The robot kinematics and dynamics were modelled. A number of control schemes were implemented on the test-bed including force proportional to velocity and acceleration; these schemes were evaluated with two subjects. PMID- 11436270 TI - The design of a five-degree-of-freedom powered orthosis for the upper limb. AB - In response to the need for a sophisticated powered upper-limb orthosis for use by people with disabilities and/or limb weakness or injury, the MULOS (motorized upper-limb orthotic system) has been developed. This is a five-degree-of-freedom electrically powered device having three degrees of freedom at the shoulder, one at the elbow and one to provide pronation/supination. The shoulder mechanism consists of a serial linkage having an equivalent centre of rotation close to that of the anatomical shoulder; this is a self-contained module in which power transmission is provided by tensioned cables. The elbow and pronation/supination modules are also self-contained. The system has been designed to operate under three modes of control: 1. As an assistive robot attached directly to the arm to provide controlled movements for people with severe disability. In this case, it can be operated by a variety of control interfaces, including a specially designed five-degree-of-freedom joystick. 2. Continuous passive motion for the therapy of joints after injury. The trajectory of the joints is selected by 'walk through' programming and can be replayed for a given number of cycles at a chosen speed. 3. As an exercise device to provide strengthening exercises for elderly people or those recovering from injury or surgery. This mode has not been fully implemented at this stage. In assistive mode, prototype testing has demonstrated that the system can provide the movements required for a range of simple tasks and, in continuous passive motion (CPM) mode, the programming system has been successfully implemented. Great attention has been paid to all aspects of safety. Future work is required to identify problems of operation, and to develop new control interfaces. PMID- 11436271 TI - MANUS--a wheelchair-mounted rehabilitation robot. AB - Rehabilitation robots are assistive devices designed for use by people with severe disability in order to gain independence in tasks of daily living. MANUS is a wheelchair-mounted general-purpose manipulator now in use with over 100 people in their homes in the Netherlands, in France and in other countries. MANUS has six main degrees of freedom, excluding the gripper. To keep the arm slim, the motors are placed in the main base. The resulting low mass of the arm ensures a reduced impulse in case of collision. A complex system of toothed belts, hollow shafts and geared wheels drives the end-effector. MANUS is equipped with a two fingered gripper. The fingers have passive canting mechanisms which ensure a three-point grasping action for most objects. A modular control system is used for steering the manipulator to its desired position. For this, a simple but effective user interface has been developed which is configurable by the end-user as well as by the therapist. PMID- 11436272 TI - The development of the Edinburgh modular arm system. AB - Modularity has not been investigated in any significant way in the development of upper-limb prostheses. The components currently available are the results of different research programmes conducted at different times by a variety of academic and commercial teams. The result is a historical hodge-podge of systems which are largely mutually exclusive in terms of compatibility. The Edinburgh work seeks to solve this problem by designing components which are neutral in structural terms so that left- and right-handed prostheses can be configured from the same basic parts. Currently, child and adult components are separate items; this does not need to be the case and the design presented here allows adult elbow parts to be used as children's shoulders. This paper will cover the rationale behind the design values and the technical aspects of the development. PMID- 11436273 TI - Three-dimensional shoulder kinematics in individuals with C5-C6 spinal cord injury. AB - The shoulder kinematics of five able-bodied subjects and those of five arms in three subjects with spinal cord injuries at C5 or C6 levels were measured as the subjects elevated their arms in three different planes: coronal, scapular and sagittal. The range of humeral elevation was significantly reduced in all spinal cord injury (SCI) subjects relative to able-bodied subjects. Over this restricted range of humeral motion, the scapula of SCI subjects tended to be medially rotated, relative to able-bodied subjects, and the protraction and spinal tilt angles of the scapula of the SCI subjects indicated scapular winging. These results are consistent with paralysis or at least with significant weakness of the serratus anterior muscle. If further study confirms this hypothesis, functional neuromuscular stimulation of the serratus anterior muscle via a nerve cuff electrode may be an effective intervention for improving shoulder function in C5-C6 SCI. PMID- 11436274 TI - Reciprocating gait prosthesis for the bilateral hip disarticulation amputee. AB - This paper describes the design, manufacture and use of a new prosthesis which produced reciprocating gait for a bilateral hip disarticulation amputee. A special mechanism used the hip extension moment during weight bearing to drive the contralateral limb through the swing phase. The user rapidly attained efficient and safe reciprocating gait, together with simple donning and doffing. The success of this project has major implications for hip disarticulation amputees in terms of profound improvements of their independence and self-esteem. PMID- 11436275 TI - Learning motor control in the upright position: a mechanical engineering approach. AB - Motor control problems that result from damage to the central nervous system can make it difficult to learn functional activities. Although several therapeutic approaches attempt to address such problems there is little evidence of their efficacy. Targeted Training is a new approach, based on a biomechanical analysis of the problem, which usually relies on specifically designed equipment. This paper describes the underlying theory, the development of functional specifications, the translation of these specifications into an engineering design and the clinical results of applying the equipment. The promotion of head and trunk control provides an example of the process. PMID- 11436276 TI - The application of the neural network on Morse code recognition for users with physical impairments. AB - Morse code is a simple, speedy and low cost means of communication composed of a series of dots, dashes and space intervals. Each tone element (either a dot, dash or space interval) is transmitted by sending a signal for a defined length of time. This poses a challenge as the automatic recognition of Morse code is dependent upon maintaining a stable typing rate. In this paper, a suitable adaptive automatic recognition method, combining the least-mean-square (LMS) algorithm with a neural network, was applied to this problem. The method presented in this paper is divided into five modules: space recognition, tone recognition, learning process, adaptive processing and character recognition. Statistical analyses demonstrated that the proposed method elicited a better recognition rate in comparison with other methods in the literature. PMID- 11436277 TI - Special issue on rehabilitation engineering. PMID- 11436278 TI - [Primary gastrointestinal lymphomas]. PMID- 11436279 TI - [Clinical features of gastrointestinal lymphoma: comparison between low-grade MALT lymphoma and high-grade lymphoma]. PMID- 11436280 TI - [Intestinal permeability in Crohn's disease and effects of elemental dietary therapy]. AB - Enteral intake of non-metabolic monosacharide and disaccharide, followed by measurement of the urinary excretion ratio of the two, is a method used to investigate intestinal permeability. L/R ratio (lactulose/1-rhamnose urinary excretion ratio) is considered an indicator of permeability of the small intestine. An increased L/R ratio is caused by mucosal disorders of the small intestine. The L/R ratio in all patients (n = 92) with Crohn's disease was 0.079 +/- 0.081 (mean +/- S.D.), which was significantly higher than the value in normal controls (0.027 +/- 0.009, n = 20, p < 0.05). In 39 patients with Crohn's disease, we assessed intestinal permeability before after treatment with an elemental diet, and during remission. The L/R ratio was 0.120 +/- 0.092, before treatment and 0.065 +/- 0.097 after treatment (p < 0.05), showing increased intestinal permeability before elemental dietary treatment. During remission, the L/R ratio was 0.035 +/- 0.028; this did not differ significantly from the value obtained after treatment. We conclude that intestinal permeability is useful for investigating disease activity in patients with Crohn's disease. PMID- 11436281 TI - [A case of ulcerative colitis associated with Graves' disease]. PMID- 11436282 TI - [A case of successful mesalazine enema for rectal cavitating ulcer of Crohn's disease]. PMID- 11436283 TI - [A case of multiple colonic ulcerations and megacolon associated with nonsteroidal antiinflammatory drugs]. PMID- 11436284 TI - [A case of hepatocellular carcinoma growing in bile duct: usefulness of enhanced power doppler ultrasonography]. PMID- 11436285 TI - [A case of small cell carcinoma of the pancreas]. PMID- 11436286 TI - [A case of malignant mesothelioma of the pleura and peritoneum with high serum levels of CA15-3]. PMID- 11436287 TI - [Gene amplification induced by the replication fork barrier site in yeast]. PMID- 11436288 TI - [DNA homologous recombination and recombinational repair mediated by RecA/Rad51 family proteins]. PMID- 11436289 TI - [Analyses of recA Rad51-like gene deficient mice]. PMID- 11436290 TI - [Functional regulation of the Mre11/Rad50/Xrs2 complex in the processes of recombination and DSB repair]. PMID- 11436291 TI - [The gene for Nijmegen breakage syndrome and its function in rejoining double strand break]. PMID- 11436292 TI - [Homologous DNA recombination is essential for the proliferation of vertebrate cells]. PMID- 11436293 TI - [Roles of BRCA1 and BRCA2 in DNA repair and tumor suppression]. PMID- 11436294 TI - [Illegitimate recombination mediated by DNA double strand break and end-joining]. PMID- 11436295 TI - [Biological functions of DNA helicase responsible for Werner syndrome]. PMID- 11436296 TI - [Function of RecQ family helicases and Bloom's syndrome]. PMID- 11436297 TI - [DNA double-strand break repair and radiation effects]. PMID- 11436298 TI - [Translesion synthesis defective in the xeroderma pigmentosum variant cells]. PMID- 11436299 TI - [DNA polymerase zeta (zeta) and error-prone translesion replication]. PMID- 11436300 TI - [Tolerance of DNA damage and RAD6 epistasis group]. PMID- 11436301 TI - [Molecular mechanism of mismatch repair]. PMID- 11436302 TI - [Mismatch-repair system and tumorigenesis]. PMID- 11436303 TI - [Oxidative DNA damage]. PMID- 11436304 TI - [Molecular mechanisms of generation and correction of DNA replication error]. PMID- 11436305 TI - [Mutagenesis by Escherichia coli DinB and its mammalian homolog Pol kappa]. PMID- 11436306 TI - [MTH1 functions in the prevention of the spontaneous mutations]. PMID- 11436307 TI - [In vivo mutation analysis with transgenic mouse]. PMID- 11436308 TI - [Human hereditary disorders caused by expansion of triplet repeats]. PMID- 11436309 TI - [Activation of p53 protein after DNA damages]. PMID- 11436310 TI - [Roles of ATM in DNA damage checkpoint and telomere maintenance]. PMID- 11436311 TI - [DNA polymerase epsilon and relative factors in the S phase checkpoint]. PMID- 11436312 TI - [Cell cycle checkpoint mechanism enforced by kinases hChk1 and hCds1]. PMID- 11436313 TI - [Role of the RFC-related Rad24 protein in the DNA damage checkpoint pathway]. PMID- 11436314 TI - [Molecular mechanism of mammalian nucleotide excision repair]. PMID- 11436315 TI - [Function of XPG gene and symptoms caused by its defect]. PMID- 11436316 TI - [Transcription-coupled repair and Cockayne syndrome]. PMID- 11436317 TI - [Base excision repair: DNA glycosylase and AP endonuclease]. PMID- 11436318 TI - [DNA repair of oxidative damage in the mitochondria]. PMID- 11436319 TI - [Repair of alkylated DNA and consequences of its failure]. PMID- 11436320 TI - [Biological significance of base damage and its repair in brain and neurons]. PMID- 11436321 TI - [DNA photolyase/cryptochrome protein family]. PMID- 11436322 TI - [Repair networks for base damage and single-strand breaks]. PMID- 11436323 TI - [Structure and function of DNA repair enzyme UvrB from Thermus thermophilus HB8]. PMID- 11436324 TI - [Structural basis for the recognition and removal of damaged bases from DNA by members of a DNA glycosylase superfamily]. PMID- 11436325 TI - [Structure determination of a DNA-repair protein by NMR]. PMID- 11436326 TI - [Roles of the Holliday intermediate in DNA recombination repair]. PMID- 11436327 TI - [Cigarette smoking and bronchial asthma]. PMID- 11436328 TI - [Chemokines in allergy]. PMID- 11436329 TI - [An evaluation of the long-term effect of clustered immunotherapy in house-dust mite-sensitive adult bronchial asthma]. AB - The optimal treatment period for specific immunotherapy in atopic asthma remains to be established. The present study was undertaken to evaluate the efficacy and the optimal treatment period of immunotherapy in adult asthma patients. Eight house-dust-mite-sensitive asthmatics were initiated by clustered immunotherapy (C IT) using house-dust (HD) antigen followed by maintenance treatment with 10-fold dilution of HD. The increase in morning peak expiratory flow rate reached a plateau at 16 weeks following C-IT. On the other hand, the maximal improvement in clinical scores was observed at a year following C-IT. Thus, it would be reasonable to determine the clinical response of immunotherapy at a year following maintenance treatment. PMID- 11436330 TI - [The diagnostic significance of immediate and late-phase skin reactions in patients with atopic dermatitis in early infancy]. AB - The diagnostic significance of immediate reaction (IR) and late-phase skin reaction (LPR) in early infancy was evaluated. Twenty six infants aged 3 months, with atopic dermatitis, received an intracutaneous test injection of egg white extract or control solution. Total serum IgE levels, peripheral eosinophile counts, CAP-RAST scores, and lymphocyte stimulation tests (LST) with egg white extract were measured. Infants were tested and followed every 3 months up to 12 months of age. There was a significant relationship between the diameter of IR elicited by intracutaneous injection of egg white extract and total serum IgE levels (p < 0.01). The diameters of IR and the diameters of LPR did not correlate. In some cases, flares over 10 mm appeared 15 minutes after injection of control solution. The average serum IgE level in the IR-positive group tended to be higher than that in the IR-negative group but this difference was not statistically significant. The stimulation index (SI) of LST in the LPR-positive group was significantly higher than that in the control group (p < 0.05). In the RAST-positive group more cases had the acute symptoms provoked by egg ingestion at 12 months of age as compared to the RAST-negative group (p < 0.05). And in the IR-positive group also more cases had the symptoms than in the IR-negative group (p < 0.01). For infants aged 3 and 6 months the number of cases with positive RAST scores following injection of egg white extract was significantly higher in the IR-positive group than in the IR-negative group (p < 0.005, p < 0.05). There was no significant difference of the RAST positive ratio between the LPR-positive and the LPR-negative group. At 6 months of age, there were more cases with positive RAST scores in the IR- and/or LPR-positive group than in the group negative for both (p < 0.05). In the LPR-positive group more cases had symptoms of allergic bronchitis or bronchial asthma before 12 months of age as compared to the LPR-negative group (p < 0.005). PMID- 11436331 TI - [An electron spin resonance (ESR) study of lymphocyte membrane fluidity in patients with bronchial asthma and atopic dermatitis]. AB - The lymphocyte membrane fluidity of patients with allergic diseases was measured by electron spin resonance (ESR), and the effect of the epinephrine stimulation on the membrane fluidity was examined. The peripheral lymphocytes were obtained from 15 patients with bronchial asthma and atopic dermatitis (28.7 +/- 9.9 years old, 10 females and 5 males) and 11 healthy adults (30.5 +/- 4.6 years old, 2 females and 9 males). Lymphocyte membranes were spin-labeled with 5-doxyl-stearic acid. Before and after the stimulation of epinephrine of the final concentrations at 10(-5) and 10(-4) mol/l, ESR spectra of the outer membranes were analyzed to evaluate the membrane fluidity. The membrane fluidity of the intact lymphocytes of allergic patients was significantly decreased in comparison to healthy controls. Although the epinephrine stimulation increased the lymphocyte membrane fluidity, the increase in fluidity was less in allergic patients than in healthy controls. There are various receptors on the surface of the lymphocyte membranes, and changes of the membrane fluidity have an influence on their functions. The results in this study elucidate the decreased fluidity of lymphocyte membrane in patients with bronchial asthma and atopic dermatitis, and suggest that the functions of the membrane receptors might be impaired. PMID- 11436332 TI - [The weather chart pattern inducing asthma attack: the advocacy of "fine mist" as a provocative factor]. AB - Outbreaks of asthma attack on 345 patients were studied in relation to the atmospheric phenomena. About 76% of days in which multiple attacks took place were fit with either of the following atmospheric conditions, 1) the trough of atmospheric pressure, 2) the approach of Typhoon, 3) the cold advection, in Japan. The common feature of these different atmospheric conditions are the tendency to turn excess vapor into fine water particles (fine mist). The fine mist could stimulate irritable airway to asthma attacks ("fine mist" hypothesis). To verity this hypothesis each asthmatic attack was further studied. It fitted in 70.7% of the attacks (fitting group) and not in 29.3% of the attacks (non-fitting group). Frequency of asthmatic attacks varied in every month on fitting group, but was nearly constant on non-fitting group. The average of the daily change of vapor density was calculated monthly. And it appeared that its graphic pattern was similar to the graphic pattern of asthmatic attack frequency. Thus it is supposed that there are at least two factors that could induce asthmatic attacks, one would be fine mist, others are unknown. PMID- 11436333 TI - [Pollen surveys in Sapporo for 6 years during 1992 to 1997]. AB - We carried out pollen surveys for 6 years from 1992 to 1997. The method of pollen survey studied was based on the standardization board of air born pollen survey and pollen information in Japan. The pollen samples were collected in Nishioka area of Toyohira-ku, and Kawazoe-area of Minami-ku in Sapporo. As the pollen sample device. IS Rotary pollen trap was used at Nishioka-area during 1992 to 1994 and at Kawazoe-area in 1993, and Durham's pollen trap was used at Kawazoe area during 1995 to 1997. The yearly pollen counts were highly fluctuated in trees, particularly Yew (Taxus spp.), Alder (Alnus spp.), Birch (Betula spp.), Pine (Pinus spp.), Fir (Abies spp.), than in Grasses (Gramineae) and Weeds (Polygonaceae, Plantago spp., Artemisia spp., Chenopodiaceae). In addition, the starting date of pollination was widely fluctuated year by year in trees. For instance, the pollen of Birch was confirmed on April 13 in 1993, as the first day in Nishioka-area, while it was on April 28 in 1994. It should be considered that the pollen counts and the starting date of pollination were highly dependent on the weather condition of the previous year and the current year. PMID- 11436334 TI - [The pollen survey and dynamic statistics of patients with allergic rhinitis in Hakodate]. AB - We have investigated the pollen survey (1994-1998) and dynamic statistics of patients with allergic rhinitis (1999-2000) in Hakodate, which is located southern part of Hokkaido. We have noted the pollen dispersion of Cryptomeria japonica, Cupressaceae, white birch, Gramineae and Artemisia. Especially, a lot of dispersion of Cryptomeria japonica has been noted in April. Concerning the dynamic statistics of patients with allergic rhinitis, we have investigated the 192 patients with allergic rhinitis in Hakodate municipal hospital. There has been a lot of pollinosis in March, April, May and September. Frequency of positive reaction to the specific IgE have been 38.0% of house dust, 16.9% of Artemisia, 13.2% of Gramineae, 10.3% of white birch, 9.0% of Cryptomeria japonica and 6.9% of cat in 379 subjects. In conclusion, we have noted that Cryptomeria japonica and white birch in addition to Gramineae and Artemisia are becoming more important antigen in patients with pollinosis in Hakodate, south part of Hokkaido. PMID- 11436335 TI - [Hypothermia in neurological diseases]. PMID- 11436336 TI - [Emerging strategies for the treatment of acute ischemic stroke: lesson from focal ischemia model]. PMID- 11436337 TI - [Magnetic resonance imaging findings of postresuscitation encephalopathy: sequential change and correlation with clinical outcome]. AB - OBJECTIVE: The purpose of this study was to describe the findings of sequential magnetic resonance imaging (MRI) in postresuscitation encephalopathy. Although its outcome is known to be overwhelming, but its acute findings by variable imaging methods are subtle and show only limited values. The correlation of the findings of MRI with clinical outcome were also analyzed. METHODS: Twelve patients with global cerebral anoxia who underwent MRI with conventional and diffusion-weighted imaging were enrolled in this study. Compared with normal MRI images, abnormal signal regions were checked and described in cortex, basal ganglia and white matter. Also medical records were carefully reviewed to study the cause, the time necessary for resuscitation and long term clinical outcome. RESULTS: The earliest finding was obtained by diffusion-weighted image less than 24 hours (acute period) in bilateral cerebral cortex as bright high signal intensity regions. Similar abnormality of bright high signal area in FLAIR and T 2 was followed according to the time elapsed in early subacute period (1-13 days). Succeedingly, white matter was involved and laminar necrosis in cortical area was observed in late subacute period (14-20 days). Finally, diffuse brain atrophy and obtundation of gray-white matter junction were seen in chronic stage (after 21 days). These MR findings were coincided well with histopathological findings reported in literatures. The poor outcome was closely and significantly correlated with abnormality in MR images. CONCLUSION: MRI was a useful diagnostic modality to diagnose the whole brain ischemic encephalopathy and to predict the prognosis. PMID- 11436338 TI - [Study on the latency difference between compound muscle and sensory nerve action potentials]. AB - In motor nerve conduction studies compound muscle action potentials (CMAPs) appear later than sensory nerve action potentials (SNAPs). This time lag originates from the conduction delay at the distal motor axon, neuromuscular transmission time and muscle action potential induction time. To investigate the latency difference between CMAPs and SNAPs we studied 46 healthy individuals, 46 patients with diabetes mellitus and 33 patients with carpal tunnel syndrome, using the lumbrical and interossei recording method. In this method the recording active electrode was placed on the 2nd lumbrical muscle and the reference electrode on the proximal palmar aspect of the index finger. Supramaximal stimulation was given to the median or ulnar nerve trunk at 9-cm proximal to the recording active electrode. The CMAP from the 2nd lumbrical muscle (L) and the SNAP from the digital nerve (N) were recorded after median nerve stimulation, and the CMAP from the 2nd interossei muscles (I) was recorded after ulnar nerve stimulation. The residual latency, which is arbitrary defined as the latency difference (L-N) in this study, was 1.38 +/- 0.15 (mean +/- SD) msec in healthy individuals. About 1 msec of the residual latency is regarded as the time for neuromuscular transmission and the time to evoke muscle activities. Thus, the conduction delay at the distal motor axon was calculated as about 0.4 msec in healthy individuals. The residual latency was relatively constant in 29 diabetic patients without conduction delay across the carpal tunnel, which was defined by the latency difference (L-I) < or = 0.4 msec. Their sensory nerve conduction velocities (calculated from N latency) were always above 40 m/sec. On the other hand in diabetic patients with conduction delay across the carpal tunnel, which was defined by the latency difference (L-I) > 0.4 msec, the residual latency gradually increased as the sensory nerve conduction velocity decreased. Their sensory nerve conduction velocities were mostly less than 40 m/sec. The similar relationship was observed in patients with carpal tunnel syndrome without diabetes mellitus. We consider that the diabetic neuropathy alone doesn't cause the increase of the residual latency. Instead, severe conduction delay across the carpal tunnel decreases the N velocity and increases the residual latency. We can also regard the relationship between the latency difference (L-N) and N velocity as being in inverse proportion. Perhaps the increase of the residual latency was simply caused by the proportional decrease in the conduction velocity at the distal motor axon, not by the special mechanism concerning to the carpal tunnel syndrome. This paper presented the electrophysiological changes seen in the distal segment secondary to the proximal entrapment. PMID- 11436339 TI - [Therapeutic outcome of spasmodic torticollis]. AB - We investigated 117 patients with spasmodic torticollis who had visited us to seek for appropriate treatment in these 14 years. They were 71 men and 46 women, aged 44 +/- 14 (mean +/- SD) years, and suffered from this disorder during 4 +/- 5 years, maximum 26 years. Involuntary abnormal head positions, not only torticollis but also laterocollis and antero- or retrocollis, were contained in this study. Most of them were torticollis due to idiopathic focal dystonia. One or more courses of alcoholization therapy was accomplished in 82 patients who wished to be done. This therapy course consisted of about ten times totally of 99% ethanol injection to the motor point of two most hypertonic neck muscles, either side of the sternocleidomastoideus and the opposite side of the splenius in most cases, repeated every 2 or 3 weeks. One patient received as many as 98 times of this injection and resolved completely. Training to reinforce antagonistic muscles was also instructed. Twenty-one patients (26%) were resolved completely after this treatment. Fifty-four patients (66%) were ameliorated and satisfied partially, but 18 of them relapsed in 1 to 4 years after the treatment and were obliged to repeat one more course of this treatment. On the other hand, in five patients their torticollis improved under certain drug therapy alone. Sixteen patients (14%) gave up to continue the treatment within two months, and 14 patients (12%) dropped out before starting the therapy. This alcoholization therapy resulted in amelioration of torticollis in about 90% of the patients with a long effective period. Nevertheless, this alcohol injection is painful, and requires 5 to 6 months to be completed. In 2 patients who had already received many times of this injection, sudden hoarseness occurred one day immediately after the alcohol injection to the sternocleidomastoideus. This complication was presumably brought about by the unexpected infiltration of alcohol to the laryngeal area, located posterior to that muscle. They recovered in two months, but careful attention should be paid to the adverse effects. If botulinum toxin be available also in our country, we will be able to have another choice of therapy and the treatment of this disorder will become easier. PMID- 11436340 TI - [Effects of preserved erythrocytes and activated-preserved erythrocytes with phosphoenolpyruvate on energy metabolism in rat ischemic brain tissue]. AB - The effects of erythrocyte function on cerebral ischemia were studied from the perspective of filtration capability (deformability) and total oxygen delivery. Erythrocytes of Wistar rats were divided into three groups: fresh blood, preserved blood, and activated preserved blood (preserved erythrocytes in which total oxygen delivery was increased with phosphoenolpyruvate). Filtration capability was calculated and evaluated by a filtration technique, while total oxygen delivery was evaluated by calculating the efficiency of oxygen delivery from an oxygen dissociation curve. Exchange transfusion was performed in spontaneously hypertensive rats with each of these groups at the same hemoglobin concentration. Bilateral carotid arteries were then ligated for one hour, and, cerebral blood flow and concentrations of adenine nucleotide and lactic acid in cerebral tissue were measured. The filtration capability of both preserved blood and activated preserved blood decreased to about one-third that of fresh blood. The efficiency of oxygen delivery after blood transfusion decreased to 17% with preserved blood, which was about half that (35%) with fresh blood. In the group of with activated preserved blood, this rate increased to 52%. The adenylate energy charge (E.C.) in brain tissue markedly decreased to 0.27 with preserved blood, which was much lower than the value (0.74) with fresh blood. With activated preserved blood, the E.C. value was 0.39. Thus, the decrease in E.C. was mild. With regard to the lactic acid concentration in brain tissue, the value with fresh blood was 12 mumoles/g tissue, while that with preserved blood was 24 mumoles/g tissue, and that with activated preserved blood was 18 mumoles/g tissue. With respect to cerebral blood flow, the values with preserved blood and activated preserved blood decreased to about one-half that with fresh blood. These results suggest that an increase in total oxygen delivery by erythrocytes contributes to improving energy metabolism in brain tissue during cerebral ischemia, and particularly during poor microcirculation. PMID- 11436341 TI - [Non-invasive examinations successfully select patients with medial temporal lobe epilepsy for anterior temporal lobectomy]. AB - We retrospectively analyzed 8 patients with intractable medial temporal lobe epilepsy (MTLE) who underwent the anterior temporal lobectomy with hippocampectomy (ATL) without invasive examinations such as chronic subdural electrode recording. Five patients had a history of febrile convulsion. While all 8 patients had oral automatism, automatism of ipsilateral limbs with dystonic posture of contralateral limbs was demonstrated in 2 patients. Bilateral temporal paroxysmal activities on interictal EEG was observed in 4 patients and all patients had clear ictal onset zone on unilateral anterior temporal region. MRI demonstrated unilateral hippocampal sclerosis in 5 cases. Interictal FDG-PET depicted hypometabolism of the unilateral temporal lobe in all cases, however, ECD-SPECT failed to reveal the hypoperfusion of the unilateral temporal lobe in a case. Postoperatively, 7 cases became seizure free, and one had rare seizure. Non invasive examinations, especially ictal EEG and concordant FDG-PET findings, in patients with oral automatism in seizure semiology, successfully select patients with MTLE for ATL. PMID- 11436342 TI - [A case report of mirror writing with low perfusion of bilateral anterior cerebral arteries]. AB - A 20-year-old female experienced temporary unintentional mirror writing associated with low perfusion of the bilateral anterior cerebral arteries. When she was 17 years old, she had developed multiple idiopathic intracerebral hemorrhages and right hemiparesis. At the age of 20, she had a generalized convulsion for which she was transferred to our department. Computed tomography (CT) and magnetic resonance images of the brain were obtained, but no fresh abnormal lesion could be detected. The following day, after she had recovered from postictal symptoms, she wrote mirror image words, and her mirror writing then gradually improved within one week. Single photon emission CT showed low perfusion of both anterior cerebral arteries. We concluded that bilateral vascular insufficiency to the supplementary motor areas and corpus callosum caused mirror writing in this case. PMID- 11436343 TI - [Atypical Fisher syndrome with optic nerve involvement]. AB - We report a 38-year-old woman who developed what appeared to be Fisher syndrome associated with optic nerve involvement. One week after a common cold, she developed double vision and left facial palsy. Four days after the onset, she developed bilateral blurred vision, painful total ophthalmoplegia, and ataxic gait. Brain CT and MRI findings were normal. Her vision worsened but the optic fundi were normal. Serum anti-GQ 1 b antibody was elevated. She received steroid therapy at another hospital, and her vision, facial palsy and ocular pain improved. She was transferred to our hospital and we treated her by plasma exchange. She showed near complete recovery. Human optic and ocular nerves contain high amount of GQ 1 b. This may be a underlying mechanism for optic nerve involvement in Fisher syndrome. We thought that she had an atypical Fisher syndrome associated with optic nerve involvement. PMID- 11436344 TI - [A case of visual agnosia for picture with right occipital lobe infarction]. AB - We report a 74-year-old right-handed man with visual agnosia for picture due to right occipital lobe infarction. The patient had a remarkable impairment in visual recognition for standardized pictures made by Snodgrass and Vanderwart, in addition to left hemianopsia, left visuospatial neglect, and mild prosopagnosia. The visual agnosia for picture was generally recognized as a mild-type of the visual object agnosia, which was extremely rare in the patients with right occipital lesion. We discussed the mechanism of the visual agnosia in the right occipital lesion. Therefore, it raises the possibility that the broad impairment of the right occipital artery territory including parahippocampal gyrus as well as corpus callosum can cause the visual agnosia for picture. PMID- 11436345 TI - [MRI findings of acute disseminated encephalomyelitis associated with mycoplasma infection]. PMID- 11436346 TI - [Teaching employees]. AB - I explained stress education for employees around the following four points based on Guidelines for Mental Health Promotion at the Workplace. That is, understanding to employee's stress, the necessity for self-care, awareness of their stress condition and how to cope with a stress condition. Finally, the content (Introduction of relaxation, stress-check by using questionnaires, case studies and Q and A) of the stress education which I was doing was presented. PMID- 11436347 TI - [Psychological counseling as a management of stress in work place]. AB - We have made psychological counseling service a part of a mental health program for 10,700 employees since 1980. Our counseling method is based on a psychological stress model and divided into three steps. The first is psychological stress survey by means of the Job Stress Scale (JSS). JSS was administered to a group of about 500 employees. The result of the JSS is reported to each employee by in-house letter. The second is a substructured interview for maladjusted employees found in JSS psychological stress reactions scores. The interviewer explains each of the JSS sub-scales such as job stressors, coping strategies, social support, and psychological stress reactions to interviewees. The last is psychological counseling. Counselees are chosen by the interviewer according to the following conditions: 1) request for counseling, 2) need for psychiatric treatment, 3) strong psychological stress reactions in spite of low job stressors, 4) signs of family problems. These four steps may decrease an employee's resistance to counseling. PMID- 11436348 TI - [A survey on occupational health nursing activities and evaluation in TOHOKU area]. AB - We did a mail survey in 1998 in Tohoku district. We sent questionnaires to approximately 300 Occupational Health (OH) nurses and got 132 replies. Occupations of respondents were 66 public health nurses and 57 nurses. More than 70% did not have a managerial position. About 40% were respondents without colleagues in nursing occupations. Full time occupational physicians were in 40% of companies, and semi-full time occupational physicians were in 60%. Respondents citing good coordination in the former were 60%, and in the latter were 80%. Of all business activities occupying OH nursing employees, persons who performed more than 90% of the tasks numbered more than 30%. Persons not satisfied with present employment positions were 80%. Reasons for dissatisfaction in declining order of incidence were work duties, contract conditions for employment, and personal relations. Planning and summary of OH nursing activities were carefully done highly. For the OH nursing activity evaluation, we examined guidelines for business locations by observing them from the OH nursing aspect and the OH nursing job itself. Guidelines considered for business locales from the OH nursing vantage point show numerically, for example, rates of examinees with abnormal findings, work absenteeism, etc, and expectations as to whether or not health conditions are conspicuously reflected in productivity. Conversely, from the OH nursing vantage point for guidelines on business locales, while receiving affirmative economic evaluation of occupational health and safety measures, progress (process) should also be included in the object of evaluation. Furthermore, guidelines should be mindful of the need to have qualitative and quantitative changes in health behavior of workers and perspectives on health. PMID- 11436349 TI - A new strategy for cancer therapy based on a predictive indicator. AB - Tumor-associated genes have been analyzed at the molecular level in recent years, and using the analyses of these genes as a predictive indicator for cancer therapy has attracted attention. Among such genes, the actions of a tumor suppressor gene p53 are focused on the cancer therapy, and it is suggested that p53 genotypes can be used as a predictive indicator for radiotherapy, thermotherapy and chemotherapy. Transfection of wtp53 to p53-null cells increased radiation- or thermo-sensitivity and stimulated apoptosis induced by these therapies. Although therapy-induced apoptosis is suppressed in mp53 cells, apoptosis can be stimulated by glycerol treatment as a chemical chaperone. Therefore, a new strategy of combining p53-targeted gene therapy or chemical chaperone therapies is expected to improve the outcome and efficiency of cancer therapy in the future. PMID- 11436350 TI - Expanded polyglutamine stretches lead to aberrant transcriptional regulation in polyglutamine diseases. AB - At least nine neurodegenerative diseases are known to be caused by expanded CAG repeats encoding polyglutamine (polyQ) stretches. Although cytotoxicities of expanded polyQ stretches have been suggested, the molecular mechanisms of neurodegeneration remain unclear. We demonstrated that the nuclear translocation of mutant proteins containing expanded polyQ stretches is a prerequisite for the expression of their cytotoxicity. Hypothesizing that nuclear proteins that interact with mutant proteins, particularly, those that bind to the expanded polyQ stretches, are involved in the pathogenetic mechanisms underlying neurodegeneration, we screened nuclear proteins for their capability of binding to expanded polyQ stretches. We found that expanded polyQ stretches preferentially bind to TAF[symbol: see text]130, a coactivator involved in CREB dependent transcriptional activation. The binding of TAF[symbol: see text]130 with expanded polyQ stretches strongly suppress CREB-dependent transcriptional activation, suggesting that interference with transcription due to the binding of expanded polyQ stretches with TAF[symbol: see text]130 and redistribution of TAF[symbol: see text]130 are involved in the pathogenetic mechanisms underlying neurodegeneration. PMID- 11436351 TI - Gene therapy with adenovirus-mediated glial cell line-derived neurotrophic factor and neural stem cells activation after ischemic brain injury. AB - Recent advancements in molecular biology are made to expect the appearance of the new treatment of stroke patients. One is the administration of neurotrophic factors, and another is the use of neural stem cell. In this report, we performed two experiments. First experiment is administration of glial cell line-derived neurotrophic factor (GDNF) using an adenovirus vector into ischemic rat brain. A replication-defective adenoviral vector containing GDNF gene (Ad-GDNF) was directly injected into the cerebral cortex at 1 day before 90 min of transient middle cerebral artery occlusion (MCAO) in rats. Infarct volume of the Ad-GDNF injected group at 24 h after the transient MCAO was significantly smaller than that of vehicle or Ad-LacZ treated group. These results suggest that the successful exogenous GDNF gene transfer ameliorates the ischemic brain injury after transient MCAO in association with the reduction of apoptotic signals. Second one is the neural stem cell activation after transient ischemia. We investigated a possible expression of highly polysialylated neural cell adhesion molecule (PSA-NCAM) in gerbil hippocampus after 5 min of transient global ischemia in association to the proliferation of neural stem cell labeled with bromodeoxyuridine (BrdU). The number of PSA-NCAM positive cells increased in dentate gyrus (DG) at 10 and 20 days, and that of BrdU-labeled cells increased in DG at 5 and 10 days after the reperfusion. Immunofluorescence for PSA-NCAM and BrdU showed that a few cells per section were double labeled in DG only at 10 days after the reperfusion. These results suggest different chronological change of PSA-NCAM positive and BrdU-labeled cells in DG after transient ischemia. PMID- 11436352 TI - Gene therapy for Parkinson's disease: determining the genes necessary for optimal dopamine replacement in rat models. AB - This article reviews the mechanism of dopamine delivery in the CNS in order to determine the optimal set of genes for effective gene therapy in Parkinson's disease (PD). Systematic neurobiological investigation of the biochemical steps has revealed that tyrosine hydroxylase (TH), which has been used in earlier studies, functions only when the essential cofactor, tetrahydrobiopterin (BH1) is present. Transduction of the gene for GTP cyclohydrolase I, the first and rate limiting step in BH1 synthesis, along with the TH gene, generated cells that are capable of producing L-DOPA spontaneously both in vitro and in vivo. When the aromatic L-amino acid decarboxylase (AADC) gene was added as a third gene, in an attempt to increase the conversion of L-DOPA to dopamine, feedback inhibition by the end product, dopamine, on TH activity resulted. To circumvent this problem, we employed a complementary strategy. Gene transfer of the vesicular monoamine transporter was combined with AADC and produced genetically modified cells that can convert L-DOPA to dopamine and store it for gradual release. This approach provided a means to regulate final dopamine delivery by controlling precursor doses and to achieve more sustained delivery of dopamine. Our investigation into determining the genes necessary for optimal dopamine delivery has been facilitated by in vivo biochemical assays using microdialysis. This technique has provided us with a clear and quantitative tool to compare the effects of various genes involved in dopamine synthesis and processing. PMID- 11436353 TI - Treatment of glioblastoma by direct inoculation of concentrated high titer recombinant retrovirus carrying the herpes simplex virus thymidine kinase gene. AB - We prepared retroviruses carrying the lacZ gene or herpes simplex virus thymidine kinase (HTK) gene with titers of 1.4-2.5 x 10(11) colony-forming units (cfu)/ml, and stereotaxically inoculated only 3 microliters of the retroviruses into a mouse glioma model. This resulted in highly efficient transduction in vivo. The transduced glioma cells migrated far from the implantation site, potentiating the induction of the remarkable bystander effect. Following repetitive ganciclovir (GCV) intraperitoneal injection, effective killing of glioma cells in the mouse brain was observed. The transduction efficiency was nearly as high as that observed for the implantation of high-titer retrovirus-producing fibroblasts. Eighty per cent of brain tumor-bearing mice were completely cured by our treatment protocol using concentrated HTK-harboring retroviruses. Our results suggest that repeated inoculations of high-titer retroviruses carrying the HTK gene followed by GCV treatment may be a promising strategy for the clinical treatment of malignant gliomas. To achieve further safety in the gene therapy of glioma, genes abundantly expressed in human glioblastoma were searched by the Serial Analysis of Gene Expression (SAGE) technique. Among the top-147 most expressed tags in glioblastoma, we found a tag, TTTTGGGTAT, originated from an unidentified gene, which was not detected in human astrocyte cultures. Real-time quantitative RT-PCR showed that MAGE-E1 expression was 2.6-15 fold enriched in glioblastoma relative to human astrocytes. Expressed Sequence Tags (ESTs) containing this tag were homologous to melanoma-associated antigen gene (MAGE) family, and this new cDNA, named MAGE-E1, was cloned by 5'-rapid amplification of cDNA ends (RACE) technique. MAGE-E1 expression was enriched in glioblastoma and low in other cancers, and MAGE-E1 expression was detected only in brain and ovary among normal tissues. These results indicate that MAGE-E1 is a novel and glioma specific member of MAGE family, which can be applied to glioma-specific gene transduction. PMID- 11436354 TI - Migration pathways and behavior of glial progenitors in the postnatal forebrain. AB - The postnatal subventricular zone (SVZ) gives rise to many of the glial cells in the forebrain. We investigated migration pathways and dynamics of motility of progenitors from the neonatal rat forebrain SVZ by labeling progenitors in vivo with a retrovirus encoding green fluorescent protein (GFP) and then visualizing the dynamics of their movements by time-lapse fluorescence microscopy in slice preparations. Cells within the dorso-lateral SVZ moved in an apparently undirected fashion, but migrated in a directed manner after emigration into white matter and cortex, displaying both radial and tangential migration. Cells in the striatal-SVZ, a region of SVZ along the lateral wall of the ventricle, migrated parallel to the ventricular surface, and entered the striatum, where they migrated both perpendicular and parallel to the ventricular surface. Sometimes, cells in all these regions reversed their migration back toward the SVZ. Migration involved either elongation of the leading process followed by a quick translocation of the nucleus or a synchronous advancement of the nucleus and the leading process. Two distinct patterns of cellular changes were observed at orthogonal turning: one involves the cessation of cell body movement and the formation of a new leading process, and the other involves continuous cell body movement and bending of the leading process. The dynamic behavior of progenitors may reflect local tissue architecture and contribute to the widespread distribution of glia. PMID- 11436355 TI - Adenoviral gene transfer of glial cell line-derived neurotrophic factor to injured adult motoneurons. AB - Glial cell line-derived neurotrophic factor (GDNF) strongly supports the survival of injured neonatal motoneurons, suggesting its potential uses in the treatment of motoneuron injury and motor neuron diseases. We examined neuroprotective effects of an adenoviral vector encoding GDNF (AxCAhGDNF) on the survival of lesioned adult rat facial and spinal motoneurons. The facial nerve or the seventh cervical segment (C7) ventral and dorsal roots of 3 month-old Fischer 344 male rats were avulsed and removed from the stylomastoid or vertebral foramen, respectively, and AxCALacZ (adenovirus containing beta-galactosidase gene), AxCAhGDNF, or PBS was inoculated in the lesioned foramen. One week after the avulsion and treatment with AxCALacZ, the animal showed expression of beta galactosidase activity in lesioned facial and spinal motoneurons. Animals avulsed and treated with AxCAhGDNF showed intense immunolabeling for GDNF in lesioned facial and spinal motoneurons and expression of virus-induced human GDNF mRNA transcripts in the lesioned brain stem and spinal cord tissues. The treatment with AxCAhGDNF after avulsion significantly prevented the loss of lesioned facial and C7 spinal motoneurons, ameliorated choline acetyltransferase immunoreactivity, and suppressed the activity of nitric oxide synthase in these neurons. These results indicate that the adenovirus-mediated gene transfer of GDNF may prevent the degeneration of motoneurons in adult humans with motoneuron injury and motor neuron diseases. PMID- 11436356 TI - NG2 cells in the brain: a novel glial cell population. AB - There exists a significantly large population of glial cells in the mammalian central nervous system (CNS) that can be identified by the expression of the NG2 proteoglycan. Cells that express NG2 (NG2 cells) are found in the developing and mature CNS and are distinct from neurons, astrocytes, microglia, and mature oligodendrocytes. They are often referred to as oligodendrocyte progenitor cells because of their ability to differentiate into oligodendrocytes in culture. However, the observation that a large number of NG2 cells persist uniformly and ubiquitously in the adult CNS and display a differentiated morphology is not entirely consistent with the notion that NG2 cells are all oligodendrocyte progenitor cells. The role of NG2 cells in oligodendrocyte regeneration and their non-progenitor role in the mature CNS are discussed in this review. PMID- 11436357 TI - Pericellular activation of hepatocyte growth factor/scatter factor (HGF/SF) in colorectal carcinomas: roles of HGF activator (HGFA) and HGFA inhibitor type 1 (HAI-1). AB - Activation of hepatocyte growth factor/scatter factor (HGF/SF) is a critical limiting step in the HGF/SF-induced signaling pathway mediated by MET receptor tyrosine kinase. Although HGF/SF-MET signaling could have potentially important roles in the invasive growth of tumors and tumor angiogenesis, little is known about the regulation of HGF/SF activation in the tumor tissues. This activation occurs in the extracellular milieu caused by proteolytic cleavage at the bond between Arg194-Val195 in the single-chain HGF precursor to generate the active two-chain heterodimeric form. Here we show that activation of HGF/SF is significantly enhanced in colorectal carcinoma tissues compared with normal colorectal mucosa, and HGF activator (HGFA), a recently identified factor XII like serine proteinase, is critically involved in this process. Furthermore, we also show that HGF activator inhibitor type 1 (HAI-1) should have an important regulatory role in the pericellular activation of HGF/SF having diverse roles acting as a cell surface specific inhibitor of active HGFA and a reservoir of this enzyme on the cell surface. The latter property might paradoxically ensure the concentrated pericellular HGFA activity in certain cellular conditions in which shedding of HAI-1/HGFA complex from the plasma membrane is upregulated. PMID- 11436358 TI - Expression of enzymes and oncogene induced after radiotherapy and/or chemotherapy in patients with brain tumors. AB - The relationship between the degree of the expression of Cu/Zn SOD, GST-pi and bcl-2 in the initial and recurrent tumor tissue after radiotherapy and/or chemotherapy and the cellular heterogeneity obtained from DNA content by image cytometry was investigated. Subjects were 7 patients who had glial tumors which were surgically removed at onset and removed a second time at recurrence. Radiotherapy and chemotherapy were also administered after initial resection. Immunoreactivity for copper/zinc super oxide dismutase (Cu/Zn SOD), GST (glutathione-S-transferase)-pi, and bcl-2 were evaluated from routinely prepared tissue blocks. Tumors were classified into two groups by cytometric analysis of DNA ploidy in the G2M cell cycle phase. One tumor group consisted of single clonal cells in both the initial and recurrent tumors and the other group consisted of tumors with polyclonal cells in the initial and recurrent tumor. In this study, one patient (case 3) with single clonal cell glioblastoma at recurrence did not show high Cu/Zn SOD activity after radiotherapy and chemotherapy but showed a short survival time after recurrence. In three patients (cases 1, 2, 3) with single clonal-cell glioblastoma, the recurrent tumor cells showed high GST-pi immunoreactivity and survival time was short after recurrence. Tumor cells in two patients (cases 5, 7) with single clonal cell anaplastic glioma at recurrence, showed high GST-pi immunoreactivity and had a short survival time after recurrence. In three single clonal glioblastomas (cases 1, 2, 3), the recurrent tumor showed the increased bcl-2 immunoreactivity and showed a short survival time after recurrence. In two patients (case 5, 7) with single clonal cell anaplastic glioma at recurrence, tumor cells showed a high bcl-2 immunoreactivity and these patients showed a short survival time after recurrence. Although the number of subjects is very small, our study shows that the immunoreactivity of bcl-2 and GST-pi in malignant gliomas may be very important factors in radio- and chemosensitivity, and shows that GST-pi is induced by radiation and anti-cancer drugs. PMID- 11436359 TI - Reduction of serum interleukin-5 levels reflect clinical improvement in patients with atopic dermatitis. AB - Cytokines, in particular IL-4 and IL-5, regulate IgE synthesis and eosinophil activation in atopic dermatitis (AD). To elucidate whether the serum levels of IL 4 and IL-5 are related to the serum IgE level, eosinophilia, or clinical severity of the disease, 25 cases with AD were studied. Blood samples were isolated from two groups of donors: 1) patients with AD (n = 25); 2) non-allergic individuals (NA, n = 20) with serum IgE levels below 100 IU/ml and with blood eosinophil counts below 250/microliter. Each parameter was evaluated at least twice in AD patients at the beginning of the study and after 4, 8 or 12 weeks of treatment. IL-4 was hardly detected in AD and NA, but IL-5 was increased (> 10 pg/ml) in most cases (22/25) of AD group with 513.6 pg/ml as the mean. AD with normal serum IgE levels exhibited increased levels of IL-5, whereas AD with high serum IgE levels did not necessarily have elevated IL-5 levels. The IL-5 level tended to change in parallel with the clinical severity in each AD case, although the level itself was not correlated with the clinical severity per se. A significant decrease of IL-5 was observed in AD when the clinical severity decreased. Eosinophils also decreased along with the improvement of AD, whereas the serum level of IgE did not change during the observation period. Our results suggest that IL-5 is involved in the regulation of clinical courses of AD and that its kinetics at the serum level reflects the clinical activity of AD. PMID- 11436360 TI - Changes in the seasonal dependence of atopic dermatitis in Japan. AB - Although it has been generally accepted that seasonal aggravation of skin symptoms is a basic feature of atopic dermatitis, recent studies have suggested a decrease in the seasonal dependence of the dermatosis in European countries over the past several decades. We examined incidence of seasonal deterioration of atopic dermatitis in Japanese subjects. The study population consisted of 682 patients aged 3 to 30 years with atopic dermatitis, who visited our outpatient clinic from August 1998 through July 2000. Of these, 452 (66%) showed a seasonal aggravation of skin symptoms, and 230 (34%) had perennial deterioration. The overall incidences of exacerbation in spring, summer, autumn and winter were 25%, 19%, 11% and 36%, respectively. Further, 358 (52%) showed unpredictable and irregular exacerbations of skin symptoms every few days or weeks. These irregular worsenings were more frequent in patients who showed perennial deterioration than in those with seasonal aggravation. From a comparison of the present results with those reported in the 1960s, it became evident that a real decrease in the incidence of seasonal aggravation of atopic dermatitis has occurred over the past three decades in Japan and that the incidence of summer deterioration has greatly decreased during the same period of time. PMID- 11436361 TI - Acute urticaria with elevated circulating interleukin-6 is resistant to anti histamine treatment. AB - Histamine released from dermal mast cells plays a central role in the increased vascular permeability in acute urticaria, and administration of anti-histamines usually suppresses development of wheals. Acute idiopathic urticaria, particularly a severe case, occasionally presents with acute inflammatory reactions such as low-grade fever and leukocytosis and is resistant to anti histamines. Considering the wide spectrum of proinflammatory cytokines and chemokines that can be released from activated mast cells, some of them might be involved in the pathogenesis of urticaria. We measured plasma levels of interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF alpha) in 16 cases of severe acute urticaria. None of them showed elevated plasma levels of IL-8 or TNF-alpha. Nine out of 16 acute urticaria patients showed elevated circulating IL-6 with concomitant increases in serum CRP levels. All such patients were resistant to conventional anti-histamine treatment and required systemic steroids for complete suppression of wheal development. After subsidence of the urticaria, their elevated IL-6 and CRP levels dropped to their normal ranges. In contrast, all but one patient without elevated circulating IL-6 was successfully treated with a H1 receptor antagonist, cetirizine. The data suggest involvement of IL-6 in the pathogenesis of severe acute urticaria that is resistant to anti-histamines. PMID- 11436362 TI - Cutaneous leiomyomatosis with type 2 segmental involvement. AB - A 21-year-old man had histologically-confirmed diffuse cutaneous leiomyomatosis. The lesions showed a peculiar distribution in that they predominantly involved several segments of the right side of his body; in addition, less extensive, nonsegmental lesions were present on both sides of the body. Although this case was apparently sporadic, the genetic mechanism of loss of heterozygosity provides a plausible explanation for this unusual presentation. If the patient were heterozygous for the underlying mutation, at an early developmental stage a postzygotic event of loss of heterozygosity would have given rise to a type 2 segmental involvement, resulting in pronounced lesions superimposed on the disseminated tumors of the ordinary phenotype. PMID- 11436363 TI - Palpable purpura at the site of previous herpes zoster in association with mixed cryoglobulinemia and hepatitis C virus infection. AB - A 70-year-old woman affected with chronic active hepatitis C and mixed cryoglobulinemia presented a palpable purpura on her abdominal skin in a metameric configuration, fourteen months after a herpes zoster in the same site. Histopathology showed a small vessel leukocytoclastic vasculitis in the superficial dermis. Post-zoster eruptions are variable, and their spectrum is still expanding, although the pathogenesis remains to be elucidated. Perhaps our case represents an isomorphic reaction, because this palpable purpura, probably related to HCV infection, occurred several months after herpes zoster. PMID- 11436364 TI - Complex ocular choristomas in linear nevus sebaceus syndrome: a report of two cases. AB - Two cases of linear nevus sebaceus syndrome (LNSS) are described in which ipsilateral facial nevus of Jadassohn was associated with complex ocular choristoma. One patient also had scleral osteomas, a rare occurrence in LNSS. PMID- 11436365 TI - Six cases of metastatic malignant melanoma with apparently occult primary lesions. AB - We report here six cases of malignant melanoma in which metastatic lesions were detected first. Of these, two cases showed some peculiar features: one exhibited widespread subcutaneous bleeding, probably due to venous rupture, and the other case had a rare primary lesion on the penis. In the Japanese literature, there have been 46 cases of malignant melanoma in which metastatic lesions were detected prior to the initial ones. The preferential site for metastasis was the lymph node (32 cases). The primary lesion was unknown in 35 patients. The outcomes were available for 36 of the 46 patients; 23 died, including 18 who died within two years. PMID- 11436366 TI - Calciphylaxis in a patient with end-stage renal disease. AB - Calciphylaxis is a rare and life-threatening condition of progressive cutaneous necrosis secondary to small and medium-sized vessel calcification. It is seen almost exclusively in patients with end-stage renal disease and secondary hyperparathyroidism. We experienced a case of 67-year-old man with calciphylaxis that manifested with characteristic skin lesions, pathologic findings, and laboratory changes. His skin lesions began as painful erythematous patches and subsequently progressed to necrotic ulcers with eschars on the distal aspect of the extremities. Pathologically, calcification was found in small and medium sized blood vessels in the deep dermis and subcutaneous tissue. His serum calcium was 9.5 mg/dL, phosphorus was 7.8 mg/dL, and nPTH was 99.9 pg/mL. The patient had been treated with surgical debridement and other supportive treatment. However, he eventually underwent an amputation below the right knee and died from sepsis. PMID- 11436367 TI - A suspected case of primary cutaneous actinomycosis on the buttock. AB - Primary cutaneous actinomycosis is rare and can mimic mycetoma, tuberculosis and nocardiosis. The results of cultures (aerobic and anaerobic) are not always positive. A high degree of clinical suspicion should be entertained and a combination regimen of penicillins with sulphonamides should be used for treatment. We report a patient with primary cutaneous actinomycosis over the buttock, who was successfully treated with amoxicillin and sulphonamides. PMID- 11436368 TI - Acute hemorrhagic edema in a four-year-old Japanese boy. AB - A case of acute hemorrhagic edema (AHE), a unique form of leukocytoclastic vasculitis limited only to the skin, was seen in a four-year-old boy. It was characterized by an abrupt onset of fever, peripheral edema, and rosette-shaped purpuric plaques. Although the skin lesion might be confused clinically with other dermatoses such as Henoch-Schonlein purpura, Sweet's disease and erythema multiforme, his laboratory data were normal, and the disorder follows a benign course with spontaneous resolution within one week. PMID- 11436369 TI - Generalized hyperpigmentation of the skin due to vitamin B12 deficiency. AB - A 49-year-old man presented with neurosis, hyperpigmentation of the skin, and depigmentation of the hair. On examination, hyperpigmentation was observed on the oral mucosa and the skin of the forearms, elbows, palmar creases and periunguinal area, knees, and feet. He had megaloblastic anemia with a low serum level of vitamin B12 due to malabsorption resulting from a gastrectomy 10 years previously. His hyperpigmentation was resolved with vitamin B12 supplementation. Histology showed an increase of melanin in the basal layer. In electron microscopic study, many melanosomes were observed in melanocytes and surrounding keratinocytes. We consider that the dominant mechanism of hyperpigmentation due to vitamin B12 deficiency is not a defect in melanin transport but is rather an increase in melanin synthesis. PMID- 11436370 TI - A fatal case of Reye syndrome after varicella and ingestion of aspirin. PMID- 11436371 TI - Paying for the pill. PMID- 11436373 TI - The replacements. PMID- 11436372 TI - New heart, new hope. PMID- 11436375 TI - The new patient power. PMID- 11436374 TI - Your next.... PMID- 11436377 TI - Reliable sources. Where to look on the Web. PMID- 11436376 TI - Made-to-order medicine. PMID- 11436378 TI - The ultimate remote control. PMID- 11436379 TI - Have we lost the healing touch? PMID- 11436380 TI - Playing with the big boys. Small hospitals get leg up in market using joint ventures with large systems. PMID- 11436381 TI - Prescription for conflict. AMA accepts funding from drug firms for campaign. PMID- 11436382 TI - No hackers here--yet. HCFA works to keep patient data private by ensuring computer security. PMID- 11436383 TI - On the trail of a computer cure-all ... eHealth Initiative. PMID- 11436384 TI - Goodbye, HCFA. Thompson, Scully unveil revamp of agency. PMID- 11436385 TI - Healthcare's rising supermodel. Hospitalist approach growing in popularity as backers cite boost in quality. PMID- 11436386 TI - Opting out. For some systems, JCAHO's accreditation cost is more than its benefit. PMID- 11436387 TI - Root-cause trouble. Court ruling in N.J. could set precedent for making JCAHO reports public. PMID- 11436388 TI - Oregon passes nurses bill. Hospitals and nurses at odds over potential effect on staffing. PMID- 11436389 TI - On the road again. Nomadic nurses earn more, see more by staying on the move. PMID- 11436390 TI - Heavy message. Detroit-area system's ads successful, but docs, others wary of tone. PMID- 11436391 TI - Corporate democracy. Scripps Health CEO forges trust between management, medical staff leaders. PMID- 11436392 TI - Gauging a storm's intensity. Senate debate on patients' rights may not presage other discussions. PMID- 11436393 TI - Promise and peril: cancer care. PMID- 11436394 TI - An industry barometer. Dominant player Hillenbrand Industries rises and falls with hospitals' health. PMID- 11436395 TI - Speaking a different language. AMA: interpreter rule would do more harm than good. PMID- 11436396 TI - When time matters most. Group working on new stroke readiness program. PMID- 11436397 TI - Foreign assignment. Cornell to open branch in Qatar; Islamic nation to get first medical school. PMID- 11436398 TI - Web survey. May survey results: a quarter-century in healthcare. PMID- 11436399 TI - Buying on credit. Hospital systems post modest increase in their long-term liabilities. PMID- 11436400 TI - Quality through leadership. PMID- 11436401 TI - A modern solution to an age-old problem. PMID- 11436402 TI - Intermediate sanctions: IRS issues new temporary regulations. PMID- 11436403 TI - Home and community-based services for well educated older caregivers: gender differences in attitudes, barriers, and use. AB - Using a sample of 169 well educated, relatively affluent older caregivers, we examined gender differences in attitudes about home and community-based services, service use, interest in services, and barriers to service use. We found significant gender differences in two attitudinal dimensions: preference for informal care and acceptance of government services. A higher proportion of men than women would rather use community services than ask family for help. A larger percentage of women than men believed the government should provide more services. Service use was modest. On average, caregivers were about 5 times more likely to express interest in a service than to have used one. Findings suggest that greater outreach by providers may usefully address service barriers. PMID- 11436404 TI - Geriatric services offered by hospitals: predicting services by internal and external community characteristics. AB - This article explores the role of hospitals in providing geriatric services and identifies associated community and hospital characteristics. The sample is 4571 community hospitals responding to the 1995 American Hospital Association (AHA) Annual Survey. Most hospitals offer some geriatric services; the mean is 3.3. The dependent variable is an index of 13 geriatric services created from the AHA data. Independent variables are taken from the AHA survey, Area Resource File, and census data. Regression analysis explains fifteen percent of the variance. Hospital characteristics predicting provision of geriatric services are non profit tax status, hospital bed size and system membership. Community characteristics predicting provision of geriatric services included higher population density, high percentage of county aged, and county nursing facility beds. Hospital characteristics yielded slightly higher predictive ability than did community characteristics. The findings suggest policies to increase hospital leadership in providing access to senior services require a multi-dimensional approach. PMID- 11436405 TI - Depression, service utilization, and treatment costs among Medicare elderly: gender differences. AB - OBJECTIVE: To compare gender differences in mood disorders, service utilization, and health care costs among a random sample of Medicare elderly beneficiaries of Tennessee. DATA SOURCES: Medicare expenditure data from a 5% random sample of Tennessee Medicare beneficiaries (n = 35,673) were examined for 1991-1993. The physician reimbursement files provided data relative to ICD-9 diagnostic codes, physician visits, and the cost of physician services provided. Other service utilization and cost data were obtained for the sample from the outpatient, home health, skilled nursing, hospice and inpatient files. STUDY DESIGN: The dependent variables were: (i) patients with ICD-9 diagnosis for a mood disorder (major depression and other depression), (ii) service utilization (number of outpatient visits, skilled nursing visits, home health visits, physician visits, emergency visits, and inpatient days), and (iii) health care costs (dollar amount of physician cost, outpatient cost, inpatient cost, total mental health cost, total health cost, and other cost). The independent variable was gender. PRINCIPLE FINDINGS: Chi-square tests showed that among the patients with a mood disorder, females had a significantly higher incidence than males of major depression (1.3% vs. .4%, respectively, p < .001) and other depression (1.6% vs. .6%, respectively, p < .001). Further, t-test results indicated that females diagnosed with major depression utilized significantly more outpatient services than males (3.2 vs. 2.6, respectively, p < .04). Total health care costs for those with other depression were significantly higher for males than females ($15,060 vs. $10,240, respectively, p < .002). CONCLUSIONS: The results indicate that mood disorders, outpatient services, and total mental health costs are higher for females than males; however, total health care costs are higher for males than females. PMID- 11436406 TI - The Long Beach Longitudinal Study: evaluation of longitudinal effects of aging on memory and cognition. AB - This article outlines the purposes, design, and findings of the Long Beach Longitudinal Study, a multipanel study of adults aged 28 through the upper reaches of the life span. The goal of the study is to identify normal short- and long-term longitudinal changes in memory and cognition across adulthood, and to develop models to explain cognitive changes. Descriptive findings of 3- and 16 year longitudinal change patterns are identified, as well as results suggesting the role of cognitive processing resources and intellectual ability in memory change. Change patterns of the oldest-old, who differ from less elderly adults, are discussed. PMID- 11436407 TI - State long-term care screening and assessment programs. AB - Long-term care screening and assessment programs were designed by states to control long-term care costs and to prevent unnecessary institutionalization of Medicaid participants. This study reports data collected by telephone survey of state officials in all 50 states and Washington, D.C. on state variation in LTC screening and assessment programs. The majority of the state screening and assessment programs cover an array of LTC services but this has resulted in multiple separate screening programs for different long-term care services and eligibility groups. Only three states coordinated screening and assessment across long-term care programs by operating a single state administrative agency, using uniform need criteria and standard tools, and having automated databases (Arizona, Colorado, and Maine). The design and implementation of multiple and separate screening and assessment programs in most states may create potential barriers to client access, information about services and choice of services. PMID- 11436408 TI - Beyond the glitz of new equipment. PMID- 11436409 TI - Endovascular stent grafting: a review. PMID- 11436410 TI - Venous access: options, approaches and issues. AB - The placement and management of venous access devices has become a routine and large part of the vascular and interventional radiologic practice. Compared with surgical series, our technical success is better and our complication rates are lower. A dedicated team approach with patient follow-up and management is required to create and maintain a successful line service. PMID- 11436411 TI - Prostate cryotherapy: practicalities and applications from the Calgary experience. PMID- 11436412 TI - Renal angioplasty and stenting. PMID- 11436413 TI - Uterine artery embolization to treat uterine fibroids. AB - Except for choice of embolization agent, the technique of uterine artery embolization for the treatment of fibroids is similar to that performed as emergent therapy for hemorrhage. The unique and more difficult aspects of fibroid embolization relate to patient recruitment and periprocedural care. There are few procedures in interventional radiology where timely and thoughtful pre- and postprocedure care are as important to building a practice. This is because of the intense and often prolonged clinical consequences of fibroid embolization, the attitude of many gynecologists toward interventional radiology and interventions for fibroids and especially because of the patient demographic. Patients seeking fibroid embolization tend to be highly motivated, educated women in their 20s to 40s with Internet access. The interventional radiologist unable to provide adequate answers or time to this group of patients in particular will find that they are an unforgiving lot! PMID- 11436414 TI - Gunther Tulip retrievable filter: why, when and how? PMID- 11436415 TI - Superficial thrombophlebitis of the breast (Mondor's disease). PMID- 11436416 TI - A simple method to minimize spillage on retrograde examination of the bowel in patients with an ileostomy or colostomy: technical note. PMID- 11436417 TI - Epidemiology of cancer mortality. AB - The level in each sex of site-specific cancers mortality is highly variable among 40 countries worldwide and somewhat less in the EU. The mortality ratio of the country worldwide with the highest upon that of the lowest cancer rate varied from 6 to 24 times in men and 6 to 17 times in women. In the EU it ranked from 3 to 10 in men and from 2 to 9 in women. Total cancer mortality had a smaller ratio (2 to 4) suggesting external and/or internal feedback mechanisms. The changes in site-specific cancer mortality rates worldwide over the years are also markedly different. A decreasing pattern since 1980 is more frequent in stomach and rectum cancer rates in each sex, in male lung cancer and in endometrium cancer. An increasing pattern is more often seen in prostate cancer, breast cancer, female lung cancer and male colon cancer. The most significant positive correlations of cardiovascular diseases are observed with rectum cancer in each sex and with endometrium cancer. Only male lung cancer correlates significantly with cardiovascular diseases. Prostate, breast and colon cancer are not positively and significantly related to cardiovascular diseases. The comparison of cancer mortality data from Belgium, The Netherlands and Denmark between 1955 and 1993 are consistent with previous results. The reliability of cancer mortality data and the role of genetic and environmental factors are discussed in two addenda. Finally it can be concluded that colon and rectum cancer behave differently at the population level. Colorectal cancer mortality data will provide misleading epidemiological results. PMID- 11436418 TI - [Is cancer a hereditary or a degenerative disease?]. AB - All malignancies, whether they are solid tumors or leukemias, always originate from modifications of the genetic information of the cells. In most cases these changes occur in a single cell, which will then generate a whole series of malignant cells through cell division. In these cases the cancer is sporadic and therefore not hereditary. In a small percentage of the cases the DNA defects are inherited from one of the parents. All cells of the body will then carry this defect, but only a few tissues will become cancerous later in life based on this defect. These forms of cancer, such as some forms of breastca, colonca, thyroidca a.o. are hereditary. Nevertheless the malignant process will not be initiated in all individuals who carry these defects and the age at onset cannot be predicted with precision. In some malignancies, whether sporadic or hereditary, the first gene that carries a defect will be essential for normal cellular function. As a result, the malignant behavior of the cells becomes irreversible. Gradually, additional defects will accumulate in other genes in these cells leading to an increasing malignant behavior of the cancer. In other cases the consequences of the first defect remain limited but the accumulation of additional defects in other genes is facilitated resulting in an increased aggressive behavior of the cells. The environmental factors that cause or facilitate the occurrence of the first DNA defect in the cells, or that facilitate the initiation of hereditary cancers remain largely unknown. Chance plays of course an important role since we accumulate continuously defects in our DNA. Nevertheless evidence exists for the presence of components in our atmosphere, our food or beverages that facilitate the accumulation of defects in our DNA. The identification of these components provides an important starting point for improved prevention. In the mean time, making genetic tests available to individuals at risk for hereditary cancers remains an important, be it delicate task, since they can potentially influence their chances of survival. The improving insights into the mechanisms involved in the pathogenesis of cancers guarantees a better and more efficient treatment in the future. PMID- 11436419 TI - [Female genital mutilation]. AB - Female genital mutilation is a collective name for all traditional practices concerning the cutting of the female genitals. Approximately 130 million are genitally cut worldwide, more than 100 million live in Africa. Annually, 2 million girls are at risk of being circumcised. FGM is deeply rooted in culture, tradition, religion and the identity. Hence, the struggle against FGM must be a combined effort of law enforcement, targeted information- and sensibilization campaigns, education and training activities for various target groups that aim at establishing behavioural changes. PMID- 11436420 TI - Genetic immunization for Chlamydia psittaci. AB - Plasmid DNA expressing the major outer membrane protein (MOMP) of an avian Chlamydia psittaci serovar A strain, has been tested for its ability to raise an immune response and induce protection against challenge with the same serovar. A combined parenteral (intramuscular injection) and mucosal route (DNA drops administered to the nares) of DNA inoculation was compared to gene gun-based immunization. The gene gun delivery of pcDNA1/MOMP as well as the intramuscular intranasal DNA delivery primed both T-helper and B-cell memory although rMOMP expressing cells did not induce high antibody responses. Evidence for the priming of the memory was provided by the fact that the pcDNA1/MOMP inoculations raised antibodies belonging to the IgG and not IgM isotype. However, in response to challenge only 5 out of 15 vaccinated turkeys showed four fold increases in serum IgG after challenge. By contrast, evidence for the priming of T-cell memory in response to challenge was found in all vaccinated turkeys as shown by the significantly heightened proliferative responses of peripheral blood lymphocytes following vaccination. Both immunization methods produced similar serological and lymphocyte proliferative responses. Notwithstanding the immunization method, a significant level of protection was observed in all pcDNA1/MOMP immunized turkeys. The efficacy of MOMP-based DNA vaccination as a means of preventing severe clinical signs, lesions and chlamydia excretion in a turkey model of Chlamydia psittaci infection was demonstrated. PMID- 11436421 TI - Cidofovir in the treatment of HPV-associated lesions. AB - The acyclic nucleoside phosphonate cidofovir (CDV) has proved efficacious in the treatment of different clinical manifestations of HPV-induced epithelial cell proliferation. Local intratumor injections of CDV in an immunocompetent patient with hypopharyngeal/esophageal papillomatous lesions, PCR-positive for HPV types 16 and 18, resulted in a complete regression of the tumor. Similarly, CDV, injected locally in patients with recurrent respiratory papillomatosis resulted in complete disappearance or partial remission of the lesions. Recently, a child with disseminated respiratory papillomatosis was treated with systemic (intravenous) CDV, resulting in stabilization of the disease. In addition, CDV topical gel has been successfully used for the treatment of severe, relapsing anogenital HPV lesions and cervical intraepithelial neoplasia (CIN) grade III. In vitro, treatment of HPV-positive cells (compared to normal primary human keratinocytes) with CDV has resulted in a concentration- and time-dependent inhibition of cell proliferation. Different parameters of apoptosis, i.e., (i) induction of CPP32 (caspase-3) protease activity, (ii) translocation of phosphatidylserine (PS) from the inner part of the plasma membrane to the outer layer, (iii) disintegration of the nuclear matrix protein (NMP), (iv) DNA fragmentation, (v) number of cells in apoptotic phase following cell cycle analysis, showed that the mechanism of cell death following treatment with CDV is based on apoptosis. Induction of apoptosis in HPV-positive cells by CDV was associated with accumulation of the tumor suppressor proteins p53 and pRb and the cyclin-dependent kinase inhibitor p21/WAF-1. In conclusion, CDV has great potential in the treatment of severe HPV-induced proliferative lesions, either laryngeal, esophageal/pharyngeal or genital. As CDV has proved able to induce apoptosis, in a time- and concentration-dependent manner, in a number of HPV positive cell lines, the regression of papillomatous tumors observed with CDV in patients, may be due, at least in part, to the induction of apoptosis. PMID- 11436422 TI - Advances in coaching: family therapy with one person. AB - This paper describes the process of "coaching" individuals in their efforts to change themselves in the context of their nuclear and parental family systems. Although this approach is regarded as one of the major modes of intervention in family therapy, the actual methods and techniques for intervention are not widely understood. Moreover, we have expanded the Bowen approach to address powerful cultural and family life cycle influences. The goal of coaching is to help clients define themselves proactively in relationship to others in their families without emotionally cutting off or giving in. Coaching begins by training clients to become observers and researchers of their own role in the family and of family patterns of behavior. Coaching then moves to help them bring their behavior more in line with their deepest beliefs, even if this means upsetting family members by disobeying family "rules." PMID- 11436423 TI - Five types of African-American marriages. AB - This study developed a marital typology based on a nonrandom, national sample of 415 African-American couples who took the Enriching Relationship Issues, Communication and Happiness (ENRICH) marital assessment inventory. Five types of African-American marriages were identified through cluster analysis using the positive couple agreement (PCA) scores in 10 relationship domains. Relationships between marital satisfaction, marital stability, and the five marital types were then analyzed. The five types (from highest marital satisfaction to lowest) were labeled as vitalized, harmonious, traditional, conflicted, and devitalized. The results were similar to findings in studies of ethnically mixed (predominantly European-American) marital samples including the number and characteristics of marital types. PMID- 11436424 TI - Motivational interviewing as an intervention for at-risk couples. AB - Thirty-one couples participated in a pilot, motivational interviewing, intervention for at-risk couples called the Marriage Checkup (MC). The MC consisted of thorough relationship assessment and individualized feedback. It attracted substantial numbers of at-risk couples who were not otherwise seeking treatment. Partners' marital satisfaction improved significantly from pre- to post-check-up and remained improved at 1-month follow up. Partners were no longer significantly different from a non-distressed comparison group following intervention. Although not addressing the efficacy of the MC, this study supports its viability as an indicated preventive intervention with couples at-risk for severe marital distress by addressing its attractiveness, tolerability, and safety. PMID- 11436425 TI - The limits of a technical concept of a good marriage: exploring the role of virtue in communication skills. AB - Teaching communication skills is one of the most common approaches used to improve couples' relationships. These skills are typically presented as content free techniques that are value neutral. Yet, marital therapists frequently see that exercising communication skills, particularly in conflict situations, can be quite difficult, requiring personal strengths such as self-restraint, courage, generosity, justice, and good judgment. These personal strengths are virtues that are presupposed in communication skills and are necessary for their successful use. The traditional attempt to see marital therapy as value neutral has made it difficult to recognize the importance of these virtues. Therapists might be more effective if they could help couples to identify and cultivate underlying character strengths necessary for good communication. This paper presents an Aristotelian reading of virtues in marriage that can broaden our understanding of marriage and open new avenues for helping couples. PMID- 11436426 TI - Projective identification in common couple dances. AB - This paper integrates the object relations concept of projective identification and the systemic concept of marital dances to develop a more powerful model for working with more difficult and distressed couples. This integrated model explains how some couples use the defenses of splitting and projective identification to externalize and transpose internal conflicts into interpersonal conflicts in five common marital dances. The author suggests how object relations techniques can be used to interrupt the projective identifications, help clients to reown their disowned and projected affects and resolve the conflict on the intrapsychic level so that the marital dance is no longer necessary to resolve the intrapsychic issues. PMID- 11436427 TI - Examining multicultural counseling competence and race-related attitudes among white marital and family therapists. AB - This study investigates the relative contributions of social desirability attitudes, previous number of multicultural counseling courses taken, and racism and White racial identity attitudes together in predicting marital and family therapists' self-reported multicultural counseling competence. Results revealed that, when controlling for social desirability attitudes and the number of multicultural courses taken, racism and White racial identity attitudes in consort accounted for a significant amount of the variance in self-perceived multicultural counseling competence. Implications for marital and family therapy training, practice, and research are discussed. PMID- 11436428 TI - Therapist perceptions of ethnicity issues in family therapy: a qualitative inquiry. AB - A group of 29 leading family therapists presenting at a regional marriage and family therapy conference were interviewed about ethnicity issues in family therapy. Questions and discussion focused on ethnicity in the family therapy process, ethnicity issues for the family therapist, ethnicity issues for the client family, therapeutic strategies when ethnicity is an issue, and recommendations for ethnicity training and supervision of future family therapists. Qualitative methods were used to analyze the transcripts of the recorded interviews to identify themes emerging from the interviews. Results reflect multiple perspectives and approaches in the field of family therapy. PMID- 11436429 TI - Developmental lessons of seasoned marital and family therapists: a qualitative investigation. AB - Little has been published regarding the development of therapists during their professional careers. This qualitative research study explored the development of seasoned marriage and family therapists (MFTs) in order to identify important events and themes in their personal and professional lives. The main developmental theme that emerged was the integration of their personal and professional selves. Information gained from this study might be useful in understanding the self of the therapist within training and supervision and might partially provide a map of the possible transitions facing MFTs along their developmental journey. PMID- 11436430 TI - Common factors across theories of marriage and family therapy: a modified Delphi study. AB - The Delphi methodology was used to explore common factors across theories of marriage and family therapy (MFT). Leading clinicians and researchers from the American Family Therapy Academy and select faculty members from MFT training programs accredited by the Commission on Accreditation for Marriage and Family Therapy Education were asked to achieve consensus regarding common factors. Respondents were asked to report on commonalities across the various MFT theories, as well as what they personally considered to be the core ingredients of change. A final profile of items was created from the responses. Qualitative interviews were conducted with selected panelists to clarify discrepancies as well as to add meaning to the data. Implications for the training and practice of MFTs as well as direction for future research are discussed. PMID- 11436431 TI - Practitioner profiles and practice patterns for marriage and family therapists in Utah. AB - This report presents the results of a survey of practitioner profiles and practice patterns for marriage and family therapists (MFTs) in Utah. A sample of 77 clinical members of the Utah Association for Marriage and Family Therapy provided descriptive information on their demographics, training, years of experience, and specific information about their practice of MFT. The findings indicate that clinical members of the American Association for Marriage and Family Therapy in Utah are a mostly male, white, and highly educated group of practitioners who hold primary licensure in MFT and identify themselves primarily as MFTs. Similarities and differences with practice patterns research in Minnesota and 15 other states are discussed. PMID- 11436432 TI - The use of allografts in ligamentous reconstructions of the knee. AB - Our continuing experience with allografts has led us to the firm conclusion that there is a clear place for allografts in the ligamentous reconstruction of the unstable knee. Clearly, issues such as cost and availability will limit allograft usage, however concerns regarding inferior clinical results should not. Our experience and those reported by others, indicates that comparable clinical results can be obtained with the added benefit of reduced surgical morbidity. In all cases meticulous attention must be given to the surgical technique. When surgical accuracy is uniformly obtained, the graft choice becomes less important when considering the sole goal of obtaining ligamentous stability in the knee. PMID- 11436433 TI - Racial differences in newborn intensive care morbidity in Alaska. AB - Birthweight-specific neonatal mortality for Alaska Natives is higher than for non natives for the years 1987-1996. We investigated the reasons for this based on Level III Neonatal Intensive Care Unit information available from 1991-1996. We also investigated whether differences in mortality extended to measures of morbidity. There were less Native patients born at the tertiary care center for babies with birthweight < 1500 grams and 1500-2499 grams (64% for Natives and 87% for non-natives, p = .000). Differences in antenatal referral were only apparent for the population residing within the Anchorage/Mat-Su area. There were also less cesarean deliveries for Native infants that were born outside of the tertiary care center for both birthweight categories (25% for Native vs. 53% for non-native infants < 1500 grams, p = .01; 27% for Native vs. 48% for non-native infants 1500-2499 grams, p = .01). For Alaska Native babies < 1500 grams there was more necrotizing enterocolitis (13% in Native vs. 4.9% in non-native, p = .01), more severe retinopathy of prematurity (12% in Native vs. 4.6% in non native, p = .01), and more bronchopulmonary dysplasia (49% in Native vs. 34% in non-native, p = .04). For Alaska Native babies 1500-2499 grams that needed ventilatory assistance there was more intraventricular hemorrhage (19% in Native vs. 7.4% in non-native, p = .003), more severe (grade 3-4) intraventricular hemorrhage (9.5% in Native vs. 0.9% in nonnative, p = .001), and more acquired sepsis (7.1% in Native vs. 1.7% in non-native, p = .02). Differences in access to Level III perinatal care and intrapartum care (cesarean delivery rates) are likely factors that contribute to the worse outcomes in the Alaska Native population. PMID- 11436434 TI - The workload deficit and hospital doctors: looking for a manageable solution. PMID- 11436435 TI - Tropical infections can mimic common western European diseases. PMID- 11436436 TI - The use of vena caval filters. AB - Deep vein thrombosis with pulmonary embolus is a frequent event, which is usually treated by anticoagulant drug therapy alone. Caval filters tend to be used infrequently in most centres in the UK. This article intends to help hospital doctors make use of these devices appropriately. PMID- 11436437 TI - The CE mark for implantable medical devices. AB - A vital aspect of device and implant regulation is continual surveillance of performance after approval. Most permanent implants seldom reveal their strengths and weaknesses until they have been in clinical use for a number of years. It is vital that clinicians report problems to the manufacturers and regulatory authorities to enable emerging hazards to be promptly identified. PMID- 11436438 TI - Nasogastric tube insertion using flexible fibreoptic nasoendoscope. AB - The insertion of a nasogastric tube, especially a fine bore feeding tube, can occasionally be problematic because of an anatomical variation or pathology. A method of passing an nasogastric tube under direct vision using a flexible fibreoptic nasoendoscope introduced through the nostril is described, which is reserved for cases where traditional methods of insertion failed. PMID- 11436439 TI - Diagnosing slipped upper femoral epiphysis. AB - The key to diagnosing slipped upper femoral epiphysis is a high index of suspicion. This article summarizes the important clinical features of the disorder and points out the pitfalls in diagnosis. PMID- 11436440 TI - The role of neurokinin B in pre-eclampsia. AB - Pre-eclampsia, a life-threatening disease unique to pregnancy, has been called a disease of theories. To date, there has been no widely accepted predictive test or therapeutic intervention to prevent or delay pre-eclampsia. The discovery of a new placental hormone, neurokinin B, may finally help to answer some of the past mysteries. PMID- 11436441 TI - The new Mental Health Act: a guide for clinicians. AB - The White Paper proposals for a new Mental Health Act have recently been published. This article examines the controversial changes in this proposal and their implications for doctors. PMID- 11436442 TI - Capacity to consent: issues and controversies. AB - Capacity to consent has major ethical and legal implications, but it is often poorly understood and inadequately practiced. Varied literature on the subject is reviewed with a view to discuss the legal and clinical aspects and strategies to improve current practice. PMID- 11436443 TI - Career success after flexible training in psychiatry: a survey of former flexible trainees in the Thames regions. AB - This article outlines the findings of a study which indicate that flexible training in psychiatry was considered adequate to prepare doctors for both full time and part-time work and that those wishing to work part time had no difficulty in finding jobs after accreditation. However, there was evidence to suggest that part-time consultants experienced more problems and less job satisfaction and that part-time training and working may restrict opportunities for an academic career. PMID- 11436444 TI - Electrocardiogram interpretation. Case 3: Progressive conduction block. PMID- 11436445 TI - Reading and assessing qualitative research. AB - What makes doctors burn out? What is it like to have epilepsy? Why do smokers not give up? Qualitative research makes it possible to look behind the statistics and to study health and health care from the inside: to find out what it is really like for the health professionals who provide the care and for the patients on the receiving end. PMID- 11436446 TI - Writing for the popular press: to inform and entertain. AB - Many doctors write for consumer publications. Even more would like the chance to do so. This article explains how to bridge the gap between writing scientific papers and writing for the popular press, and offers realistic guidance on how to find a way into this competitive market. PMID- 11436447 TI - Abdominal actinomycosis following a forgotten intrauterine contraceptive device. PMID- 11436448 TI - Abdominal tuberculosis: the great mimic. PMID- 11436449 TI - An unusual presentation of acute cholecystitis: biliary ascariasis. PMID- 11436450 TI - A political asylum seeker with an amoebic empyema in Birmingham. PMID- 11436452 TI - The eye in Marfan's syndrome. PMID- 11436453 TI - New standards for public health. PMID- 11436455 TI - Tracheostomy in the intensive care patient: surgical or percutaneous? PMID- 11436456 TI - The future of nursing. ANA president Mary Foley shares her perspective. Interview by Peg Cushman. PMID- 11436457 TI - Home care's high-level & industrywide staffing challenges. AB - Home care executives are all too familiar with the problems of the tight labor market, affecting both professional and paraprofessional staff. The realities of the market and the extent of the problem are well documented in current literature. This article sheds some light on current initiatives aimed at both immediate and long-term solutions to the health care labor shortage. PMID- 11436458 TI - Worlds apart. A family's struggle with the nursing shortage. PMID- 11436459 TI - A state association approach to the policy and politics of the worker shortage. AB - In January 2000, the Board of Directors of the Home & Health Care Association (H&HCA), identified the issue of worker recruitment and retention as a "critical issue" in the Association's strategic plan. With a different perspective from its employer/agencies, the Association began to devote more resources (staff time) to the issue, tapping into what was already happening in the state and proposing some solutions of its own. PMID- 11436460 TI - Compensation programs mean more than money. AB - How do health care and, particularly, home care administrators use incentive programs? What are health care professionals and paraprofessional looking for? Is it money, security, or just meeting the needs of everyday life? What motivators are the key to successful compensation programs? PMID- 11436461 TI - The nurse shortage. A home care agency perspective. AB - Certified home care agencies are challenged to meet an increasing demand for services while faced with a predicted severe nursing shortage. Meeting this demand will depend on an agency's ability to recruit and retain a qualified nursing staff. Knowledge of the extent of the nursing shortage, along with information of administrators' perceptions of the problem and activities they have implemented to alleviate a shortage will provide information to meet this new challenge. PMID- 11436462 TI - HCFA responds to HAA survey & certification questions on the Medicare hospice benefit. PMID- 11436463 TI - Home care aide staffing survey results. PMID- 11436464 TI - The power of a vision.... A leader's journey. AB - Being a leader in health care today requires the ability to implement a visionary style of leadership. The visionary leader has the challenge of formulating and articulating a corporate vision that employees can buy into and work toward. PMID- 11436465 TI - Home care nursing shortage must be reversed. AB - The National Association for Home Care provided the following information and recommendations regarding the nursing shortage in recent testimony prepared for the United States Senate, Aging Subcommittee, Committee on Health, Education, Labor and Pensions. PMID- 11436466 TI - Discharging older patients from home care: who decides and when? AB - Who is involved in the decisions to discharge patients from home care and how much lead-in time for planning is observed? How does the process of discharging patients from home care differ by reason for discharge? These questions are addressed in this study of 383 Medicare-funded patients discharged from home care agencies in central Ohio. PMID- 11436467 TI - Developing solutions to the direct care worker shortage. PMID- 11436468 TI - Health systems performance--what's next? PMID- 11436469 TI - Prevalence, awareness, treatment and control of hypertension among the elderly in Bangladesh and India: a multicentre study. AB - OBJECTIVE: To evaluate the prevalence, awareness, treatment and control of hypertension among elderly individuals in Bangladesh and India. METHOD: A community-based sample of 1203 elderly individuals (670 women; mean age, 70 years) was selected using a multistage cluster sampling technique from two sites in Bangladesh and three sites in India. FINDINGS: The overall prevalence of hypertension (WHO-International Society for Hypertension criteria) was 65% (95% confidence interval = 62-67%). The prevalence was higher in urban than rural areas, but did not differ significantly between the sexes. Multiple logistic regression analyses identified a higher body mass index, higher education status and prevalent diabetes mellitus as important correlates of the prevalence of hypertension. Physical activity, rural residence, and current smoking were inversely related to the prevalence of hypertension. Among study subjects who had hypertension, 45% were aware of their condition, 40% were taking anti hypertensive medications, but only 10% achieved the level established by the US Sixth Joint National Committee on Detection, Evaluation and Treatment of Hypertension (JNC VI)/WHO criteria. A visit to a physician in the previous year, higher educational attainment and being female emerged as important correlates of hypertension awareness. CONCLUSIONS: Our findings emphasize the need to implement effective and low cost management regimens based on absolute levels of cardiovascular risk appropriate for the economic context. From a public health perspective, the only sustainable approach to the high prevalence of hypertension in the Indian subcontinent is through a strategy to reduce the average blood pressure in the population. PMID- 11436470 TI - Using lot quality assurance sampling to improve immunization coverage in Bangladesh. AB - OBJECTIVE: To determine areas of low vaccination coverage in five cities in Bangladesh (Chittagong, Dhaka, Khulna, Rajshahi, and Syedpur). METHODS: Six studies using lot quality assurance sampling were conducted between 1995 and 1997 by Basic Support for Institutionalizing Child Survival and the Bangladesh National Expanded Programme on Immunization. FINDINGS: BCG vaccination coverage was acceptable in all lots studied; however, the proportion of lots rejected because coverage of measles vaccination was low ranged from 0% of lots in Syedpur to 12% in Chittagong and 20% in Dhaka's zones 7 and 8. The proportion of lots rejected because an inadequate number of children in the sample had been fully vaccinated varied from 11% in Syedpur to 30% in Dhaka. Additionally, analysis of aggregated, weighted immunization coverage showed that there was a high BCG vaccination coverage (the first administered vaccine) and a low measles vaccination coverage (the last administered vaccine) indicating a high drop-out rate, ranging from 14% in Syedpur to 36% in Dhaka's zone 8. CONCLUSION: In Bangladesh, where resources are limited, results from surveys using lot quality assurance sampling enabled managers of the National Expanded Programme on Immunization to identify areas with poor vaccination coverage. Those areas were targeted to receive focused interventions to improve coverage. Since this sampling method requires only a small sample size and was easy for staff to use, it is feasible for routine monitoring of vaccination coverage. PMID- 11436471 TI - Estimating the prevalence of anaemia: a comparison of three methods. AB - OBJECTIVE: To determine the most effective method for analysing haemoglobin concentrations in large surveys in remote areas, and to compare two methods (indirect cyanmethaemoglobin and HemoCue) with the conventional method (direct cyanmethaemoglobin). METHODS: Samples of venous and capillary blood from 121 mothers in Indonesia were compared using all three methods. FINDINGS: When the indirect cyanmethaemoglobin method was used the prevalence of anaemia was 31-38%. When the direct cyanmethaemoglobin or HemoCue method was used the prevalence was 14-18%. Indirect measurement of cyanmethaemoglobin had the highest coefficient of variation and the largest standard deviation of the difference between the first and second assessment of the same blood sample (10-12 g/l indirect measurement vs 4 g/l direct measurement). In comparison with direct cyanmethaemoglobin measurement of venous blood, HemoCue had the highest sensitivity (82.4%) and specificity (94.2%) when used for venous blood. CONCLUSIONS: Where field conditions and local resources allow it, haemoglobin concentration should be assessed with the direct cyanmethaemoglobin method, the gold standard. However, the HemoCue method can be used for surveys involving different laboratories or which are conducted in relatively remote areas. In very hot and humid climates, HemoCue microcuvettes should be discarded if not used within a few days of opening the container containing the cuvettes. PMID- 11436472 TI - Client satisfaction and quality of health care in rural Bangladesh. AB - OBJECTIVE: To assess user expectations and degree of client satisfaction and quality of health care provided in rural Bangladesh. METHODS: A total of 1913 persons chosen by systematic random sampling were successfully interviewed immediately after having received care in government health facilities. FINDINGS: The most powerful predictor for client satisfaction with the government services was provider behaviour, especially respect and politeness. For patients this aspect was much more important than the technical competence of the provider. Furthermore, a reduction in waiting time (on average to 30 min) was more important to clients than a prolongation of the quite short (from a medical standpoint) consultation time (on average 2 min, 22 sec), with 75% of clients being satisfied. Waiting time, which was about double at outreach services than that at fixed services, was the only element with which users of outreach services were dissatisfied. CONCLUSIONS: This study underscores that client satisfaction is determined by the cultural background of the people. It shows the dilemma that, though optimally care should be capable of meeting both medical and psychosocial needs, in reality care that meets all medical needs may fail to meet the client's emotional or social needs. Conversely, care that meets psychosocial needs may leave the clients medically at risk. It seems important that developing countries promoting client-oriented health services should carry out more in depth research on the determinants of client satisfaction in the respective culture. PMID- 11436473 TI - Evidence of behaviour change following a hygiene promotion programme in Burkina Faso. AB - OBJECTIVES: To determine whether a large, 3-year hygiene promotion programme in Bobo-Dioulasso, Burkina Faso, was effective in changing behaviours associated with the spread of diarrhoeal diseases. The programme was tailored to local customs, targeted specific types of behaviour, built on existing motivation for hygiene, and used locally appropriate channels of communication. METHODS: Two population surveys recorded the coverage of the programme among target audiences (mothers of children aged 0-35 months). Four surveys were carried out: three prior to the programme and one in 1998 (after the programme had been running for 3 years), using structured observation of hygiene behaviours in the participants' homes to document changes in target behaviours. FINDINGS: After the programme had run for 3 years, three-quarters of the mothers targeted had had contact with programme activities. Half could cite the two main messages of the programme correctly. Although the safe disposal of children's stools changed little between 1995 and 1998 (80% pre-intervention, 84% post-intervention), hand-washing with soap after cleaning a child's bottom rose from 13% to 31%. The proportion of mothers who washed their hands with soap after using the latrine increased from 1% to 17%. CONCLUSION: Hygiene promotion programmes can change behaviour and are more likely to be effective if they are built on local research and use locally appropriate channels of communication repeatedly and for an extended time. PMID- 11436474 TI - Group A streptococcal sore throat in a periurban population of northern India: a one-year prospective study. AB - OBJECTIVE: To estimate the incidence and risk factors of group A streptococcus (GAS) sore throat among school-aged children living in a periurban slum area of Chandigarh, North India. METHODS: A total of 536 children aged 5-15 years from 261 families identified by a systematic random selection method were enrolled in the study. Episodes of sore throat were recorded through fortnightly home visits over a one-year period. The local vernacular (Hindi) terms gala kharab (bad throat) and khansi jukam (cough and cold) were used to identify symptoms of sore throat, and throat swab specimens were collected from children who had these symptoms on the day of the home visit. Bacterial culture was carried out and the isolation of GAS was confirmed using group-A-specific antiserum. FINDINGS: The incidences of sore throat and GAS sore throat were, respectively, 7.05 and 0.95 episodes per child-year. The incidence was higher in the following situations: among 11-year-olds, during the winter (November to January) and rainy (August) months (a bimodal peak), among children living in houses where there was no separate room for the kitchen, and in homes that included a tobacco smoker. CONCLUSION: The results show that the incidence of GAS sore throat was related to age, season, and indoor air pollution. PMID- 11436475 TI - Iodine concentration in household salt in South Africa. AB - OBJECTIVE: To determine the iodine concentration in household salt, the coverage of adequately iodized salt, the use of non-iodized agricultural and producers' salt, and the usefulness of salt as a carrier of iodine, and to relate these observations to socioeconomic status in South Africa. METHOD: The iodometric titration method was used to analyse 2043 household salt samples collected using a national, multistage, stratified, cluster survey. FINDINGS: The national mean and median iodine concentrations of household salt were 27 mg/kg (95% confidence interval (CI): 25-29 mg/kg) and 30 mg/kg (range = 0-155 mg/kg), respectively. There was considerable variation within and between geographical areas. Coverage of adequately iodized household salt, i.e. iodized at > 15 mg/kg, was 62.4% of households (95% CI: 58.8-66.0%) two years after the introduction of compulsory iodization at a level of 40-60 mg/kg. A total of 7.3% of households used non iodized agricultural salt and salt obtained directly from producers. People at the lower end of the socioeconomic spectrum were more likely to suffer the consequences of using under-iodized salt because more of them used agricultural or coarse salt than did people in the higher socioeconomic categories. The iodine concentration in salt was lower in rural areas than in urban and periurban areas. CONCLUSIONS: The consequences of using under-iodized or non-iodized salt were most likely to be experienced in the country's three northern provinces, among people in the low socioeconomic categories, and in rural households. Since 95.4% of households in South Africa use salt regularly and 2.9% use it occasionally, the national iodization programme has the potential to meet the iodine requirements of the population. However, this can only be achieved if the primary reasons for the inadequate iodization of salt are eliminated and if special attention is given to vulnerable groups. PMID- 11436476 TI - Coeliac disease: a potentially treatable health problem of Saharawi refugee children. AB - OBJECTIVE: To characterize the clinical and nutritional impact of coeliac disease (gluten-sensitive enteropathy) among Saharawi children living as refugees in Algeria. METHODS: A total of 65 Saharawi children with coeliac disease were compared with 71 age-matched non-coeliac controls. For each participant, the clinical history was taken and a clinical examination, non-quantitative 24-hour dietary recall, anthropometric and skinfold measurements, bioelectric impedance analysis (BIA) of body composition, and venous blood sampling for haemoglobin determination were performed. RESULTS: Gluten-containing food, especially bread, was the staple diet of Saharawi children. Abdominal pain and distension were significantly commoner among children with coeliac disease than in controls (P < 0.05). The mean height-for-age was significantly lower in such children than in controls (-2.5 +/- 1.4 units vs -1.8 +/- 1.3 units, respectively, P < 0.01). No significant differences were found for either skinfold or BIA measurements. Haemoglobin values tended to be lower in children with coeliac disease than in controls. CONCLUSIONS: Coeliac disease has a negative effect on the health status of Saharawi refugee children. Because of the high prevalence of the condition in the Saharawi, a specific programme for treating all affected individuals should be established. Further studies are required to quantify the impact of coeliac disease in other areas of the developing world. PMID- 11436477 TI - The burden of disease in Maputo City, Mozambique: registered and autopsied deaths in 1994. AB - OBJECTIVE: To classify the causes of death in Maputo City, Mozambique, using the methods of the Global Burden of Disease study, in order to provide information for health policy-makers and to obtain a baseline for future studies in Maputo City and provincial capitals. METHODS: Data were taken from the Maputo City death register and autopsy records for 1994. FINDINGS: A total of 9011 deaths were recorded in the death register, representing a coverage of approximately 86%. Of these, 8114 deaths (92%) were classified by cause. Communicable, maternal, perinatal, and nutritional disorders accounted for 5319 deaths; noncommunicable diseases for 1834; and injuries for 961. The 10 leading causes of registered deaths were perinatal disorders (1643 deaths); malaria (928); diarrhoeal diseases (814); tuberculosis (456); lower respiratory infections (416); road-traffic accidents (371); anaemia (269); cerebrovascular diseases (269); homicide (188); and bacterial meningitis (178). CONCLUSIONS: Infectious diseases of all types, injuries, and cerebrovascular disease ranked as leading causes of death, according to both the autopsy records and the city death register. AIDS-related deaths were underreported. With HIV infection increasing rapidly, AIDS will add to the already high burden of infectious diseases and premature mortality in Maputo City. The results of the study indicate that cause of death is a useful outcome indicator for disease control programmes. PMID- 11436478 TI - On measuring inequalities in health. AB - In a recent series of papers, Murray et al. have put forward a number of important ideas regarding the measurement of inequalities in health. In this paper we agree with some of these ideas but draw attention to one key aspect of their approach--measuring inequalities on the basis of small area data--which is flawed. A numerical example is presented to illustrate the problem. An alternative approach drawing on longitudinal data is outlined, which preserves and enhances the most desirable aspects of their proposal. These include the use of a life course perspective, and the consideration of non-fatal health outcomes as well as the more usual information on mortality patterns. PMID- 11436479 TI - Maternal mortality at the end of a decade: signs of progress? AB - Maternal mortality is an important measure of women's health and indicative of the performance of health care systems. Several international conferences, most recently the Millennium Summit in 2000, have included the goal of reducing maternal mortality. However, monitoring progress towards the goal has proved to be problematic because maternal mortality is difficult to measure, especially in developing countries with weak health information and vital registration systems. This has led to interest in using alternative indicators for monitoring progress. This article examines recent trends in two indicators associated with maternal mortality: the percentage of births assisted by a skilled health care worker and rates of caesarean delivery. Globally, modest improvements in coverage of skilled care at delivery have occurred, with an average annual increase of 1.7% over the period 1989-99. Progress has been greatest in Asia, the Middle East and North Africa, with annual increases of over 2%. In sub-Saharan Africa, on the other hand, coverage has stagnated. In general, caesarean delivery rates were stable over the 1990s. Countries where rates of caesarean deliveries were the lowest- and where the needs were greatest--showed the least change. This analysis leads us to conclude that whereas there may be grounds for optimism regarding trends in maternal mortality in parts of North Africa, Latin America, Asia, and the Middle East, the situation in sub-Saharan Africa remains disquieting. PMID- 11436481 TI - Netherlands, first country to legalize euthanasia. PMID- 11436480 TI - Targeted interventions required against genital ulcers in African countries worst affected by HIV infection. AB - It remains unclear why there is such marked variation in the severity of the human immunodeficiency virus (HIV) epidemic between African countries. The prevalence of HIV infection has reached high levels in many parts of southern Africa but in most countries of West Africa the levels are much lower. Although there is good evidence that sexually transmitted infections (STIs) and genital ulcers in particular facilitate heterosexual transmission of HIV, there is little comparative STI data from the African countries worst affected by HIV infection. A MEDLINE search covering the period 1966 to August 2000 using the keywords "sexually transmitted diseases", "genital ulcers" and "Africa" was performed to identify factors that might be relevant to the spread of HIV infection in countries with the highest prevalences of the virus. In the countries worst affected by HIV infection, the proportions of men and women with STI who had genital ulcers lay in the ranges 45-68% and 13-68%, respectively. The proportions were much lower in countries of West Africa than in those of southern Africa. The African countries worst affected by HIV infection should adopt a more specialized approach to STI control than hitherto and specifically target the high incidence of genital ulceration. Locally, technical STI committees should draw up country specific guidelines taking into account the prevalence of the various causes of genital ulceration. In these countries, national AIDS control programmes and donor agencies should develop a specific focus for decreasing the incidence of genital ulcer disease. PMID- 11436482 TI - Heated debate likely on plan for EU-wide health coordination. PMID- 11436483 TI - Arsenic in water--how much is too much? PMID- 11436484 TI - Polio eradication--the last and toughest 1%. PMID- 11436485 TI - Human genome sequences--a potential treasure trove, but how useful? PMID- 11436486 TI - Measles eradication still a long way off. PMID- 11436487 TI - Vectors of diseases. Hazards and risks for travellers. Part I. PMID- 11436488 TI - Monitoring the emergence of antiretroviral resistance. PMID- 11436490 TI - Reclaiming Freud's seven articles of faith. AB - Defines "faith" as the underpinning of every therapist's choice of focus when responding to a specific client, reflecting the therapist's belief that this chosen focus will contribute most to that client's future well-being. Discusses seven of Freud's recommendations for technique as based on the faith that these will maximize future benefit to clients. "Reclaims" these approaches in Freud's name, since many modern-day techniques are reinventions of Freud's original approach. PMID- 11436491 TI - Self-reported value of spiritual issues among adolescent psychiatric inpatients. AB - Reports on a survey of a prospective, uncontrolled sample of adolescent psychiatric inpatients on the importance of spiritual issues to them. Results indicated that the vast majority of adolescents in this sample indicated that spiritual beliefs are considered important. Notes that a majority reported that mental health professionals rarely asked them about their beliefs in the spiritual areas. Observes that nearly all the patients reported having a chaplain led spiritual group that they perceived as a helpful part of the therapeutic milieu. PMID- 11436492 TI - Using a modification of the classic Drama Triangle to enhance pastoral care. AB - Describes the Drama Triangle of Victim-Persecutor-Rescuer conceptual model and suggests helpful ways to use it in order to understand and intervene in the difficult situations often encountered by pastors, pastoral counselors, and chaplains. Attempts to join this conceptual model to a theological understanding of persons to deepen self-examination, ground pastoral identity, and enhance pastoral competence. PMID- 11436493 TI - Wounding, doubting, and trusting in suffering. AB - Constructs a working model of pastoral care on the foundation of the reality of suffering. Explicates how suffering wounds the person both psychologically and spiritually and how doubting God can be a constructive phase leading to the reclaiming of trust in God. PMID- 11436494 TI - Revisiting the process theology of Daniel Day Williams with implications for pastoral care. AB - Reviews the basic assumptions and insights of Daniel Day Williams' process theology as they relate to his understanding of the human predicament and his concept of deliverance and suggests implications for contemporary pastoral care. PMID- 11436495 TI - The congregational setting of pastoral counseling: a study of pastoral counseling theorists from 1949-1999. AB - Examines, through content analysis, the major authors in the field of pastoral counseling since 1949 with the goal to determine the degree to which pastoral counseling theory in the second half of the twentieth century attended to the context of parish ministry. Reports on trends in the field's orientation and notes that a particular viewpoint, or range of viewpoints, has driven the core of pastoral counseling theory and methodology. Attempts to answer the question, Does the literature of pastoral counseling address the counseling situations typically encountered by congregational pastors and provide an adequate methodology suited to the context of parish ministry? PMID- 11436496 TI - The Muslim festival of Eid ul-Fitr at L'Arche Daybreak: a pattern and principles for common worship services in solidarity with those not of the majority faith. AB - Describes a common worship pattern designed to include a Muslim woman but a pattern that could be readily adapted in many situations of community or institutional chaplaincy settings where one or more of the members or residents is not of the majority religion. PMID- 11436497 TI - Mortality moments. PMID- 11436498 TI - Heaven is the place where nothing ever goes wrong. PMID- 11436499 TI - We're all "passing". PMID- 11436500 TI - Pastoral counseling in prison: how to be. PMID- 11436501 TI - [Antimicrobial activities of meropenem against clinically isolated strains in 1999]. AB - We determined the antibacterial activity of meropenem (MEPM) and control drugs against clinical isolates of 310 strains of Gram-positive bacteria (14 species), 590 strains of Gram-positive bacteria (21 species), and 120 strains of anaerobic bacteria (10 species) in 1999. We compared the results thus obtained with those in 1993 and 1997. The results were as follows; 1. MEPM showed excellent antibacterial activities against most of the species tested, except for MRSA, E. faecium, E. avium, and methicillin-resistant S. epidermidis (MRSE). 2. MEPM had much higher activity than IPM and PAPM against Gram-negative bacteria including S. marcescens and P. aeruginosa, part of which have been reported to produce metallo-beta-lactamase. 3. There was little difference in the susceptibility of clinical isolates to MEPM between 1993 and 1999. Thus MEPM was shown to retain its potent and broad antibacterial activity now at the same level as before available for use in 1995. PMID- 11436502 TI - [Clinical investigation of cefpirome for perinatal infection and infection prevention]. AB - Effectiveness and safety of cefpirome (CPR) were studied in perinatal infection and prevention of postoperative infections. 1. Enrolled in the study of perinatal infections were the total 62 cases comprising 47 of chorioamnionitis, 10 of puerperal intrauterine infections, 1 of infectious abortion and 4 of others. The effective rate of CPR 2 g/day given in drip infusion was 61/62 (98.4%). In bacteriological studies, the bacterial elimination rate was 57/61 (93.4%). Unchanged and remained were 3 strains of Gram-positive bacteria and 1 strain of Bacteroides spp. 2. Enrolled in the study of prevention of postoperative infections during perinatal period were the total 88 cases comprising 74 who underwent cesarean section and 14 others. The effective rate of CPR 2 g/day given in drip infusion was 87/88 (98.9%). 3. With respect to subjective and objective adverse affects due to the agent, drug eruption was observed in 1/150 (0.7%). No abnormal result of clinical laboratory tests was found. The above results suggested the usefulness of CPR for treatment of perinatal infections and prevention of postoperative infections. PMID- 11436503 TI - [Lymphodynamic factors governing lymphatic spread of carcinoma cells]. AB - Carcinoma frequently spreads and grows through the lymphatic system, whereas malignant tumors of mesenchymal origin more frequently spread via the hematogenous route. This paper reviews cancer spread with special reference to tumor microcirculation, vascular endothelial growth factor receptor (VEGFR)-3 mediated lymphangiogenesis, regeneration of collecting lymph vessels, and the role of nitric oxide in lymphatic metastasis. Oxygen and pH are key microenvironmental factors in the development and growth of tumors and their response to treatment. Thus the physiological and pathophysiological roles of hypoxia and acidic pH in the tumor microenvironment in lymphatic metastasis are demonstrated. Lymph angiogenesis in tumor tissues and regeneration of collecting lymph vessels with special reference to VEGFR-3 and its agonists VEGF-C/VEGF-D are discussed. The occurrence and biological significance of intratumoral lymph angiogenesis in breast cancer have been established. VEGF-C as a molecular link between tumor lymph angiogenesis and metastasis is identified. Finally, the role of nitric oxide (NO) in tumor microcirculation and lymphatic metastasis of carcinoma cells is evaluated. PMID- 11436504 TI - [Pathological features of lymph node metastasis. 2) From morphological aspects]. AB - Lymph node metastasis is important because of its early appearance and prognostic significance. In this paper the mechanism of development of metastasis, tissue reaction with metastatic cells, and role of lymph node metastasis in cancer spread are described briefly. Lymph node metastasis proceeds when cancer cells invade lymphatic vessels. These cells lodge in the lymphatic sinus where tumor cells adhere to and breach the sinusoidal walls and migrate into the parenchyma. Integrin(s), CD44, and/or carbohydrates of tumor cells and extracellular matrix components such as laminin and type IV collagen, hyaluronic acid, and lectins of the sinus wall become involved in the adherence of cancer cells. Their binding may induce matrix metalloproteinase production from cancer cells, resulting in degradation of surrounding reticular fibers. Lymph nodes sometimes react with tumors, exhibiting follicular hyperplasia, sinus histiocytosis, lymphoid cell depletion, fibrosis, and angiogenesis of lymph nodes. Such tissue reactions may suppress cancer spread but in turn may augment the aggressiveness of cancer cells. It is not yet possible to determine which reaction will occur. In early cancer, lymph node metastasis occurs based on mainly anatomical-mechanical factor, while in advanced cancer, seed-soil factor plays an important role on development of the metastasis. PMID- 11436505 TI - [Sentinel lymph node (SLN) in breast cancer: prediction of axillary metastasis confined to the SLN spares patients further axillary dissection]. AB - Although regional control of the axilla in patients with breast cancer is important, axillary lymph node dissection (ALND) is performed mainly for staging purposes. The sentinel lymph node (SLN) that first receives lymphatic drainage from the tumor was investigated. After SLN identification, ALND or radiation therapy is performed in patients with SLN metastasis who are also likely to have non-SLN metastasis. However, it is important to select patients with SLN metastasis who may benefit from further axillary treatment. The size of the primary tumor and the size of its SLN metastasis are reported to be associated with the presence of non-SLN metastases. Patients in whom the SLN indicates less than 20% of nodes are involved have a low probability of non-SLN involvement and therefore can avoid further axillary treatment. Although large clinical trials are necessary for confirmation, it appear, that patients with axillary involvement confined to the SLN can be spread further ALND. PMID- 11436506 TI - [Significance of detection of micrometastases in regional lymph nodes in cancer patients using RT-PCR]. AB - Lymph node metastasis is the most important predictor of prognosis, and lymph node dissection is indispensable in achieving curative resection. However, even in patients who undergo curative resection, 20-30% experience relapse, which may be due to micrometastases that are undetectable by conventional histologic methods. Recently, more specific and sensitive genetic methods, including the reverse-transcriptase polymerase chain reaction (RT-PCR), have been introduced to detect micrometastases. We review the literature to evaluate the significance of detecting micrometastases in lymph nodes by RT-PCR. Since the first study by Smith in 1991, many authors have reported molecular diagnoses in the lymph nodes, blood, and bone marrow in patients with various types of cancer using markers such as CEA and cytokeratins. Many researchers have found RT-PCR to be more sensitive than conventional histological methods in detecting lymph node metastases. Molecular diagnosis will become a powerful tool for predicting prognosis and determining who should receive postoperative adjuvant chemotherapy. However, reports on the relationship between molecular diagnosis and prognosis are still few. It will be important to clarify this relationship in large-scale clinical trials to evaluate the significance of detecting micrometastases in the lymph nodes. PMID- 11436507 TI - [Treatment of lymph node metastasis from the viewpoint of surgical oncology]. AB - We have investigated methods to predict lymph node metastasis in early gastric cancer. First, the efficacy of fluorescence in situ hybridization (FISH) using the dual-color method was evaluated as a potential marker of lymph node metastasis in 20 early gastric cancers. A significant increase in the fraction of cells with a decrease in p53 was observed in early gastric cancer compared with normal tissues. More importantly, a significant increase in the fraction of cells with p53 deletion was observed in patients with lymph node metastasis. The predictive accuracy was 45%. Second, the relationship between the degree of expression of biological markers and lymph node metastasis was examined. High expression of p27 and cyclin E had a strong correlation with lymph node metastasis. Moreover, all patients with combined high expression of p27, cyclin E, and matrix metalloproteinase-9 had lymph node metastasis. However, these represented only 21% of cases with lymph node metastasis. Difficulty in the clinical use of these biological markers to detect lymph node metastasis depends on the feedback mechanism of cell cycle regulators or heterogeneity of the lesion. These problems should be resolved in the near feature. PMID- 11436508 TI - [Appropriate extent of lymph node dissection in thyroid cancer]. AB - Surgery is not always necessary for micro cancer (less than 1.0 cm in diameter). In surgery of micro cancer, we recommended to dissect only the central compartment (pretracheal [II]/paratracheal [III] area). In patient with cancer (more than 1.0 cm in diameter), lymph nodes in the central compartment, ispirateral supraclavicular area, and jugulocarotid chain (V and VI) should be dissected. Even in incomplete surgery, lymph node dissection of the central compartment is warranted in patients with papillary cancer, because recurrence in the central compartment results in dyspnea and/or hemosptum, lowering the quality of life. PMID- 11436509 TI - [Management of axillary and internal mammary lymph nodes in primary breast cancer]. AB - Axillary lymph node dissection (ALND) is an effective staging procedure and is essential for local control of breast cancer. The regimen of the adjuvant systemic therapy is largely based on the number of nodes involved. There is as yet no evidence of survival benefit from axillary treatment by either surgery or radiotherapy, but this issue remains controversial. In general, the standard treatment of the axilla is surgical clearance of nodes from level I and II (partial ALND). If these nodes are involved, the clearance of level III nodes (complete ALND) is indispensable from the viewpoint of local control. Because a high rate of adverse events is observed, the extent of ALND should be determined by considering the balance between side effects and therapeutic benefit on a case by-case basis. For the management of internal mammary nodes, most reports on randomized trials indicate that neither surgical treatment nor radiotherapy influences survival. However, the prognostic significance of internal mammary node status is high and a selected biopsy of lymph nodes with adenopathy should be considered for staging purposes. The significance of local control in this region is still controversial at present. About 30% to 40% of all invasive breast cancers are node positive. Thus, in most cases, the potential morbidity of ALND could be avoided if the status of the axillary nodes was ascertained with a less invasive procedure. The technique of sentinel lymph node biopsy may eventually prove to decrease the need for standard ALND. The randomized trial NSABP-B32 is ongoing and the results should indicate the clinical need for ALND. PMID- 11436510 TI - [Lymph node dissection in lung cancer]. AB - The standard surgical treatment for stage I, II, and IIIa non-small cell lung cancer (NSCLC) is lobectomy with mediastinal lymph node dissection. In our series of 214 patients with N2 disease the 5-year survival rate was 23%. Systematic mediastinal lymph node dissection provides further prognostic information on the N factor. Among NSCLC patients with a pathological diagnosis of N2, there are significant differences in the 5-year survival rate based the 3 factors of clinical N factor, N1 station metastasis, and N2 station metastasis. PMID- 11436511 TI - [Appropriate extent of lymphadenectomy in esophageal cancer]. AB - Because of the high frequency and widespread distribution of lymph node metastasis in esophageal cancer, three-field dissection was introduced in the early 1980s. Although improvement in the long-term survival rate of patients who undergo three-field dissection has been reported, the clinical significance of cervical lymphadenectomy is still controversial because the effect of improved upper-mediastinal lymphadenectomy in these patients cannot be excluded. Recently, the sentinel node (SN) concept has been validated in gastrointestinal cancers including esophageal cancer using the radio-guided method. Aberrant drainage routes from the primary lesion have frequently been observed in esophageal cancer and are detectable by radio-guided SN mapping. SN navigation is useful to identify SNs as the functional first basin to evaluate the regional lymph node status and to determine the indications for extensive lymphadenectomy in patients with esophageal cancer. Individualized surgical management of patients with esophageal cancer is essential to improve the quality of life of patients and long-term results. PMID- 11436512 TI - [Optimal nodal dissection for early gastric cancer]. AB - Extensive lymphadenectomy (D2) in 295 patients with early gastric cancer (EGC) resulted in a significantly lower 10-year recurrence rate than limited lymph node dissection (D1) in 97 patients (2.1% vs. 11.9%, p < 0.005). Among node-positive patients, the recurrence rate following D2 was significantly lower than that after D1 (12.5% vs. 44.4%, p < 0.02). Among node-negative patients, there was no difference in recurrence rate between two groups (0.6% vs 3.3%, p < 0.2). These observations suggest that there are two optimal methods of node dissections in EGC surgery based on nodal status. The sentinel node concept is important to understand nodal status. In 1993, we developed intraoperative endoscopic lymphatic mapping with 2% patent blue to demonstrate the lymphatic basins in EGC. Frozen sections of the blue nodes in 203 patients had a high predictive value for nodal metastasis, with a sensitivity of 89%, specificity of 100%, and accuracy of 98%. Four false-negative cases had clinical metastasis, which was diagnosed at surgery. In 34 of 35 patients, metastatic nodes were located along the lymphatic basins. Among them, 15 patients had metastasis only in the sentinel lymph nodes. Of 5 gastric lymphatic basins, 42% of the patients had involvement of one, 47% 2, and 12% 3. These results show that each EGC has its own lymphatic basins in which metastasis can develop. The more numbers of the sections there are, the higher the likelihood of nodal metastasis. This means that each frozen section slice carries the risk of being false negative. Therefore we should always dissect the lymphatic basins even in cases with no sentinel node metastasis. In addition, patients with sentinel nodes containing metastasis should be treated with the D2 procedure. PMID- 11436513 TI - [Optimal lymph node dissection for carcinoma of the biliary tract]. AB - To clarify the optimal lymph node dissection for carcinoma of the biliary tract, we analyzed the mode of lymphatic spread in 86 resected cases with carcinoma of the gallbladder and 139 with carcinoma of the extrahepatic bile duct, and investigated long-term results after resection based on the degree of lymph node metastasis. Of the 86 patients with carcinoma of the gallbladder, 62 (72.1%) had lymph node metastasis. Patients with m and mp tumors (n = 9) had no lymph node metastasis, whereas ss tumors (n = 13) had 23.1% lymph node metastasis. Those with se, si tumors (n = 64) had greater lymph node involvement (92.2%). In 4 patients with advanced carcinomas (ss or more) who survived more than 5 years, only one (limited to periportal lymph nodes) of them had lymph node metastasis. Of the 139 patients with carcinoma of the extrahepatic bile duct, 58 (41.7%) had lymph node metastasis. There was no lymph node metastasis in 15 patients with m or fm tumors. The frequency of metastasis in the ss (n = 39) and se, si (n = 85) tumors was 17.9% and 60.0%, respectively. Twenty-four patients with advanced tumors survived more than 5 years. Curative resection was achieved in all 24 and lymph node metastasis was n0 in 19, n1 in 4 and n2 in 1 patients. Satisfactory long-term result can be achieved in carcinoma of the biliary tract after resection when lymph node metastasis is limited to nodes in the hepatoduodenal ligament. In view of our surgical results and the lymphatic drainage system of the biliary tract, systemic dissection of the regional lymph nodes, including periportal, posterior pancreato-duodenal, and celiac nodes, is necessary in patients with N0-N2 (limited to lymph nodes in the hapatoduodenal ligament) tumors in whom it contributes to good prognosis. PMID- 11436514 TI - [Optimal lymph node dissection for colorectal cancer]. AB - Previous studies on the distribution of positive lymph nodes have revealed that the colon should be resected 10 cm from the tumor on both sides and that the intermediate nodes along the main vessel should be dissected in patients with colon cancer. In rectal cancer, superior lymphatic spread along the inferior mesenteric artery (IMA) is the main metastatic route. The IMA should be dissected immediately after the bifurcation of the left colic artery, and the intermediate lymph nodes should be removed. The positive rate of the lateral lymph nodes is about 10%. The rate of local failure is high and the prognosis is poor in patients with positive lateral lymph nodes, even if the lateral lymph nodes have been dissected. However, it has been reported that lateral lymph node dissection combined with excision of the internal iliac vessels results in good disease-free survival in patients with positive lateral nodes. Therefore the indications for lateral node dissection remain controversial. Lymphatic spread into the mesorectum on the anal side has been shown to be an important factor in local failure. The mesorectum should be resected for up to 4 or 5 cm from the inferior tumor margin in middle rectal cancer, and the entire mesorectum should be removed in lower rectal cancer. Nerve tissue preserved in pelvic autonomic nerve preserving surgery contains a small amount of lymphoid tissue and lymph nodes. Therefore the extent of lymph node dissection and the area of autonomic nerves to be preserved based on tumor site or tumor penetration remain controversial. PMID- 11436515 TI - [Pharmacological and therapeutic properties of lafutidine (stogar and protecadin), a novel histamine H2 receptor antagonist with gastroprotective activity]. AB - Potent antisecretory agents, such as histamine H2-receptor antagonists and proton pump inhibitors, have achieved great improvement in peptic ulcer therapy. It has, however, been reported that incidence of ulcer relapse is high after discontinuation of these drugs. Insufficient efficacy against NSAID-induced ulcers is also critical. Lafutidine is a novel histamine H2 antagonist with gastroprotective activity. Lafutidine exhibited potent and long-lasting H2 antagonism and prolonged antisecretion. In addition, lafutidine showed a gastroprotective effect against noxious agents-induced gastric mucosal damage through capsaicin-sensitive afferent nerves. Lafutidine showed antiulcer activities against acute ulcer models, prevented gastric ulcer relapse of acetic ulcer, and accelerated the healing of indomethacin-induced antral ulcers in rats. These results suggest the advantage of the combined antisecretory and gastroprotective activities. In clinical studies, lafutidine showed prolonged antisecretion, healing effect against gastric and duodenal ulcers and gastritis, and its potency was equal or superior to that of conventional H2 antagonists. Additionally, lafutidine induced a high transition rate to the E0 stage determined by endoscopical ultrasonography, suggesting the high quality of ulcer healing. Furthermore, effectiveness of lafutidine against NSAIDs-induced ulcer was high. From these results, lafutidine is equal or superior to conventional H2 antagonists in antiulcer potency, and it may be useful for the prevention of ulcer relapse and or treatment of NSAIDs-induced gastroduodenal damage. PMID- 11436516 TI - [Anti-migraine drug sumatriptan succinate, a 5-HT1B/1D-receptor agonist]. AB - Sumatriptan succinate (SMT) was a highly specific 5-HT1-receptor agonist. It showed high affinity only for 5-HT but no affinity for other neurotransmitter receptors such as muscarinic, dopamine D1, D2, adrenergic alpha 1, alpha 2, and beta. Furthermore, it was highly selective for 5-HT1B/1D-receptor and showed no affinity for 5-HT2 and 5-HT3 receptors. SMT contracted isolated cranial arteries such as basilar, midcerebral, temporal arteries and large arteries in the dura matter, but did not contract coronary, femoral, mesenteric and other arteries. Reflecting these results, SMT induced vasoconstriction of carotid artery, but produced practically no contractile responses in the other arteries mentioned above in anaesthetized animals. These pharmacological characteristics of SMT were different from those of ergot alkaloids, current anti-migraine drugs, which contracted coronary, femoral and other arteries as well. SMT inhibited neurotransmitter release, including CGRP, from trigeminal nerve terminals. Consequently protein extravasation induced by CGRP was inhibited and neurogenic inflammation could be suppressed. It was believed that SMT showed its anti migraine activity through cranial vasoconstriction via 5-HT1B/1D receptors, since it did not show any analgesic activities. Its clinical efficacy on migraine and cluster headache had been already confirmed in about 100 western countries. Its efficacy was also shown by open trials and placebo controlled double blind tests in Japan. PMID- 11436517 TI - [Pharmacological effects of cabergoline against parkinsonism]. AB - The pharmacological effects of cabergoline, a novel ergot alkaloid, against parkinsonism were assessed by comparing its effects with those of bromocriptine and pergolide. The affinities of cabergoline and pergolide for the D2 receptor were about the same, about 7 times stronger than that of bromocriptine. The affinity of each compound for the D1 receptor was markedly lower than its affinity for the D2 receptor. However, other data suggest that cabergoline and pergolide would have D1-receptor agonist activity, whereas bromocriptine would act as a D1-receptor antagonist. In MPTP-lesioned parkinsonian monkeys, cabergoline improved motor disability, and its effect lasted longer than those of bromocriptine and pergolide. Moreover, cabergoline induced no behavioral abnormalities even though at the highest dose used, in contrast to bromocriptine and pergolide, both of which induced hyperactivity. This beneficial effect of cabergoline did not attenuate on prolonged administration. Combined treatment with a low dose of L-dopa and a low dose of cabergoline improved motor disability without inducing the hyperactivity and dyskinesia seen during treatment with L dopa alone at high doses. From these results, we suggest that cabergoline promises to be a useful anti-parkinsonian agent with a long lasting effect that survives prolonged administration and without the side effects induced by L-dopa. PMID- 11436518 TI - [Effect of olopatadine hydrochloride, a novel antiallergic agent, on the QT interval in dogs]. AB - Olopatadine hydrochloride (olopatadine), a novel antiallergic agent, is effective in the treatment of allergic rhinitis, chronic urticaria, eczema and dermatitis. It has been reported that terfenadine and astemizole cause side effects on the circulatory system such as QT prolongation followed by serious ventricular arrhythmias (torsades de pointes). To investigate the possibility of QT prolongation, we used both conscious normal dogs and hypokalemia-anesthetized dogs under two conditions: 1) olopatadine used alone and 2) olopatadine used in combination with itraconazole, the CYP3A4-inhibiting antifungal agent, in the present investigation. The group treated with terfenadine alone (30 mg/kg, p.o.) and the group treated with a combination of terfenadine (10 mg/kg, p.o.) and itraconazole (100 mg/kg, p.o.) had a significantly prolonged QT interval. On the other hand, the group treated with olopatadine alone (30 mg/kg, p.o.) and the group treated with a combination of olopatadine (30 mg/kg, p.o.) and itraconazole (100 mg/kg, p.o.) did not show any significant changes in QT interval. Moreover, olopatadine (1 and 5 mg/kg, i.v.) did not influence the QT interval in hypokalemia-anesthetized dogs. These results suggest that there is very little possibility of QT prolongation as a result of clinically used olopatadine. PMID- 11436519 TI - Cold Monday mornings prove dangerous: epidemiology of sudden cardiac death. AB - Sudden cardiac death is the leading cause of death in industrialized countries, accounting for 10 to 20% of total mortality. Several studies have demonstrated a circadian variation of sudden death with a primary peak in the morning hours after awakening and a secondary peak in the late afternoon. Weekly and seasonal variations have been observed, with more frequent occurrence of sudden death on Mondays and in the winter compared with other days of the week and seasons, respectively. These patterns of disease occurrence indicate the presence of identifiable triggering factors. Interestingly, the circadian pattern of sudden death appears to be more pronounced in older patients and to be attenuated by beta-blocker therapy. Rupture of an atherosclerotic plaque with subsequent coronary thrombosis is the most common underlying pathophysiologic mechanism of sudden death. The variation in disease occurrence may reflect endogenous physiologic rhythms and the importance of external events (e.g., exertional physical activity) that trigger changes (e.g., surges in blood pressure) that lead to coronary plaque rupture. To reduce the long-term risk of sudden death, strategies of primary and secondary prevention must be further developed. To reduce short-term risk of sudden death, patients at risk for sudden death may require additional behavior modification and pharmacological intervention. PMID- 11436520 TI - Technologic advances and program initiatives in public access defibrillation using automated external defibrillators. AB - Widespread provision of early defibrillation following cardiac arrest holds major promise for improved survival from ventricular fibrillation. The critical element in predicting a successful outcome is the rapidity with which defibrillation is achieved. A worldwide awareness of this potential and its advocacy by such organizations as the American Heart Association have been pivotal in the evolution of initiatives to make defibrillation more widely and more rapidly available. The feasibility of this initiative, known as public access defibrillation, is in large measure a direct consequence of major technologic advances in automated external defibrillators (AEDs). New low-energy waveforms with biphasic morphology have been shown to be more effective in terminating ventricular fibrillation and may do so with less myocardial injury. Placement of AEDs in a variety of nontraditional settings such as police cars, aircraft and airport terminals, and gambling casinos has been shown to yield an impressive number of survivors of cardiac arrest in ventricular fibrillation. Questions yet to be answered center on the appropriate disposition of AEDs in public access defibrillation settings, training and retraining issues, device maintenance, and collection of accurate data to document benefit and to identify areas of needed improvement or expansion of AED availability. PMID- 11436521 TI - Algorithms to analyze ventricular fibrillation signals. AB - Prediction of the success of defibrillation to avoid myocardial injury and performance feedback during CPR requires algorithms to analyze ventricular fibrillation signals. This report reviews investigations on different parameters of ventricular fibrillation electrocardiographic signals, including amplitude, frequency, bispectral analysis, amplitude spectrum area, wavelets, nonlinear dynamics, N(alpha) histograms, and combinations of several of these parameters. To date, no satisfactory methods have been found that cope with CPR artifacts and show adequate predictive power of successful defibrillation. The usual limitations of the studies are the small number of subjects, which precludes separation into training and test data. Because many investigations are animal studies of untreated short ventricular fibrillation, the results may be different for prolonged ventricular fibrillation in humans. The universality of threshold values has to be examined, and promising new parameters have to be monitored over longer time periods and analyzed for the effects of chest compressions, ventilation, and concomitant vasopressor therapy. PMID- 11436522 TI - Arginine vasopressin during cardiopulmonary resuscitation and vasodilatory shock: current experience and future perspectives. AB - Epinephrine use during cardiopulmonary resuscitation (CPR) is controversial because of its receptor-mediated adverse effects such as increased myocardial oxygen consumption, ventricular arrhythmias, ventilation-perfusion defect, postresuscitation myocardial dysfunction, ventricular arrhythmias, and cardiac failure. In the CPR laboratory, vasopressin improved vital organ blood flow, cerebral oxygen delivery, resuscitability, and neurologic recovery more than did epinephrine. In patients with out-of-hospital ventricular fibrillation, a larger proportion of patients treated with vasopressin survived 24 hours than did patients treated with epinephrine. Currently, a large trial of out-of-hospital cardiac arrest patients being treated with vasopressin versus epinephrine is ongoing in Germany, Austria, and Switzerland. The new international CPR guidelines recommend 40 U vasopressin intravenously, and 1 mg epinephrine intravenously, as equally effective for the treatment of adult patients in ventricular fibrillation; however, no recommendation for vasopressin has been made to date for adult patients with asystole and pulseless electrical activity, or in children, because of lack of clinical data. When adrenergic vasopressors were unable to maintain arterial blood pressure in patients with vasodilatory shock, continuous infusions of vasopressin (0.04-0.10 U/min) stabilized cardiocirculatory parameters and even ensured weaning from catecholamines. PMID- 11436523 TI - Mechanical advances in cardiopulmonary resuscitation. AB - Challenged by the continued high mortality rates for patients in cardiac arrest, the American Heart Association and the European Resuscitation Council developed a new set of guidelines in 2000 to help advance several new and promising cardiopulmonary resuscitation (CPR) techniques and devices. This is the first time these organizations have taken such a bold move, in part because of the poor results with standard closed-chest cardiac massage. The new techniques, interposed abdominal counterpulsation and active compression decompression CPR, each provide greater blood flow to the vital organs in animal models of CPR and lead to higher blood pressures in patients in cardiac arrest. In some clinical studies, both techniques have resulted in a significant increase in survival after cardiac arrest in comparison with standard CPR. Three of the four new CPR devices that were recommended in the new guidelines also provide superior vital organ blood flow and increased blood pressures in comparison with standard CPR. The three devices that improve the efficiency of CPR are the circumferential vest, an active compression decompression CPR device, and an inspiratory impedance valve used in combination with the active compression decompression CPR device. The fourth device type, one that compresses the thorax using an automated mechanical piston compression mechanism, was recommended to reduce the number of personnel required to perform CPR. However, no studies on the automated mechanical compression devices have showed an improvement in hemodynamic variables or survival in comparison with standard CPR. Taken together, these new technologies represent an important step forward in the evolution of CPR from a pair of hands to devices designed to enhance CPR efficiency. Each of these advances is described, and the recent literature about each of them is reviewed. PMID- 11436524 TI - Thrombolytic therapy during cardiopulmonary resuscitation and the role of coagulation activation after cardiac arrest. AB - Thrombolysis is an effective causal therapy for patients suffering from massive pulmonary embolism or acute myocardial infarction. In more than 70% of patients with cardiac arrest, one of these two diseases is the underlying cause of deterioration. Nevertheless, because of the fear of severe bleeding complications, thrombolytic therapy during cardiopulmonary resuscitation (CPR) has been contraindicated. Increasing clinical experience and data from open studies now suggest that thrombolysis during CPR can contribute to hemodynamic stabilization and survival in patients with massive pulmonary embolism and acute myocardial infarction, after conventional CPR procedures have been performed unsuccessfully. After administration of thrombolytic agents, some patients have been stabilized even after more than 90 minutes of CPR. Besides the specific causal action of thrombolytic agents at the site of pulmonary emboli and coronary thrombosis, experimental data indicate that thrombolysis during CPR can improve microcirculatory reperfusion, which may be most important in the brain. In accordance with these data, marked activation of blood coagulation without adequate activation of endogenous fibrinolysis has been demonstrated during reperfusion after cardiac arrest. Massive coagulation activation with subsequent fibrin formation is responsible for microcirculatory reperfusion disorders, and thrombolytic therapy may be indicated. However, no controlled studies are available on this therapeutic concept. Because the risk of bleeding complications is potentially associated with the administration of thrombolytic agents, although this occurs far less than anticipated, thrombolysis during CPR is presently a treatment strategy that can be performed on an individual basis. Whether thrombolysis during CPR can improve survival rates and neurologic outcomes should be addressed in randomized, controlled trials. PMID- 11436525 TI - Therapeutic hypothermia after cardiac arrest. AB - This review discusses the mechanisms of neurologic damage during and after global cerebral ischemia caused by cardiac arrest. The different pathways of membrane destruction by radicals, free fatty acids, excitatory amino acids (neurotransmitters), calcium, glucose metabolism, and oxygen availability and demand in relation to metabolic rate are briefly discussed. The main focus of this review paper, however, lies in therapeutic (resuscitative) hypothermia after cardiac arrest. Two pioneering studies of the 1950s and four recent publications (in part preliminary results of ongoing studies) in humans are discussed in detail. The conclusions are as follows: (1) hypothermia holds promise as the only specific brain therapy after cardiac arrest so far; (2) hyperthermia is not tolerable after successful resuscitation; and (3) if the ongoing European multicenter trial of hypothermia after cardiac arrest finds a significant benefit to mild hypothermia, withholding hypothermia may be ethically hard to defend. PMID- 11436526 TI - Brain energetics of cardiopulmonary cerebral resuscitation. AB - Recovery of normal brain energetic conditions during and after resuscitation from cardiac arrest is critical for survival and good neurologic outcome. This review emphasizes the glucose-driven metabolic processes during and after ischemia and on the post-resuscitation development of secondary energy derangements. It also explores some potential therapeutic interventions designed to attenuate these energy derangements. The article summarizes some bench research and is not intended to provide treatment strategies for clinical application. PMID- 11436527 TI - Cardiovascular optimization for high-risk surgery. AB - Controversy still exists concerning the strategy of targeting so-called supranormal levels of oxygen delivery as a means of improving outcome from high risk surgery. The pathogenesis of postoperative morbidity is not fully defined; however, it seems likely that the gastrointestinal tract plays a pivotal role, particularly as a source of endotoxin, which is a potent stimulator of the inflammatory response. Delayed recovery of gastrointestinal function also is a major contributor to postoperative morbidity. Demonstration of a significant reduction in gut ischemia, endotoxemia, postoperative inflammatory response, and morbidity in a high-risk general surgical population as a result of perioperative hemodynamic optimization would be a highly worthwhile study. PMID- 11436528 TI - Assessment of the microcirculatory flow in patients in the intensive care unit. AB - Various techniques have been used at bedside to assess the microcirculation of critically ill patients, including nailfold videomicroscopy, laser doppler techniques, and orthogonal polarization spectral imaging. Nailfold videomicroscopy was introduced first, but its value may be limited by the extreme sensitivity of nailfold microcirculation to external temperature or vasoconstrictive agents. Laser Doppler techniques can measure gastric or jejunal mucosal blood flow as well as skin and muscle blood flow, but do not take into account blood flow heterogeneity, a major parameter of microcirculation. The recent introduction of orthogonal polarization spectral imaging techniques allows direct visualization of microcirculation in critically ill patients, opening a new area for the investigation of the pathophysiologic processes involved in the hemodynamic alterations of shock states. PMID- 11436529 TI - Central venous oxygen saturation monitoring in the critically ill patient. AB - In the initial treatment of a critically ill patient, blood pressure, heart rate, urine output, and central venous pressure guide resuscitative efforts. Despite normalization of these variables, global tissue hypoxia may still persist and has been implicated in the development of multiorgan failure and increased mortality. Definitive management includes intensive care unit admission, pulmonary artery catheterization using mixed venous oxygen saturation (SvO2), and hemodynamic optimization. In the absence of or before definitive management, hemodynamic optimization can be performed using central venous oxygen saturation (ScvO2) as a surrogate. The physiology, technology, clinical uses, and rationale for ScvO2 monitoring are reviewed, including issues regarding physiologic equivalence to SvO2. The clinical use of ScvO2 monitoring, evidence-based outcome implications, and limitations of ScvO2 monitoring will also be examined. PMID- 11436531 TI - Bibliography. Current world literature. Cardiopulmonary resuscitation. PMID- 11436530 TI - Implications of arterial pressure variation in patients in the intensive care unit. AB - Positive-pressure ventilation alters stroke volume by transiently increasing intrathoracic pressure and thereby decreasing preload. This phasic variation in stroke volume results in a cyclic fluctuation in arterial pressure with a phase length equal to the respiratory rate. Measuring ventilation-induced arterial pressure variation allows the clinician to predict the cardiovascular response to changes in intravascular volume status. Thus, one may predict preload responsiveness because the greater the amount of ventilation-associated arterial pressure variation, the greater the patient's preload responsiveness. This variation in arterial pressure can been defined as a variation in either systolic pressure or pulse pressure. Although pulse pressure gives a clearer signal, systolic pressure variation may be easier to measure bedside without invasive hemodynamic monitoring. Newer methods of quantifying this arterial pressure variation include the respiratory systolic variation test, which can performed without an apneic baseline, and the pulse pressure variation, a potentially more accurate measure of preload responsiveness. PMID- 11436532 TI - Bibliography. Current world literature. Cardiopulmonary monitoring. PMID- 11436533 TI - Combination therapy for sarcoidosis. AB - Combination therapy has proved useful in infectious, rheumatologic, and oncologic diseases. The role of combination therapy in sarcoidosis is less defined. A stepwise approach to therapy in sarcoidosis treatment includes multiple agents, such as topical and systemic corticosteroids. The introduction of cytotoxic agents has led to the combination of these drugs with lowered doses of corticosteroids. Recently, the combination of cytotoxic and immune modifiers has been used for some cases of refractory sarcoidosis. The rationale use of combination therapy may enhance efficacy with reduced toxicity. PMID- 11436534 TI - Sampling local respiratory tract sites for inflammation. AB - Although host defense mechanisms protect the health of the respiratory tract, deficiencies or excessive host responses such as inflammation can create sino nasal and pulmonary illnesses. Analysis of expectorated secretions (sputum) is traditional, but selective site specific samples (surface washing) seem more applicable for studying illnesses such as rhinitis, bronchitis, or aveolitis. Regional samples should contain representative components (cells and proteins) that ideally reflect changes in adjacent tissue; however, biopsy of contiguous mucosa and lung parenchyma is required for a comprehensive analysis that can link surface samples and tissue pathology. Clinical strategies must be flexible so that maximal information about the host can be obtained through investigation of healthy normals, volunteer-patients with latent illness that can be induced locally and observed in situ, and patients with active illness. Implicit in studying inflammation is the adaptation of new analytic methods to surface or topical samples such as nasal, bronchial and bronchoalveolar lavage (BALF). More correlations between respective components in nasal fluids, bronchial and BAL specimens seem indicated, so that more readily available samples can be used for clinical monitoring. Moreover, further correlations between bronchial and BAL cells and immunohistochemical analysis of tissue for cells are needed. PMID- 11436535 TI - Sarcoidosis: genes and microbes--soil or seed? AB - Sarcoidosis is a multi-organ granulomatous disorder that is characterised by the accumulation of CD4+ T-lymphocytes resulting in a Th-1 type immune response. Although our understanding of the immune response in sarcoidosis has improved in recent years through studies of bronchoalveolar lavage cells and fluid, the genetic predisposition and trigger factors (and their interrelationship) remain unclear. Previous reports of familial clustering and varying prevalence of sarcoidosis in different populations suggested molecular epidemiological heterogeneity. This review focuses specifically on two pivotal areas that have been the subjects of intensive investigation recently: a) triggering by infective agents and b) host genetic susceptibility and relates these to broader issues of pathogenesis. It is concluded that one or more microbes behaving in a non infectious fashion in a genetically predisposed individual trigger the sarcoidosis granulomatous response. PMID- 11436536 TI - Analysis of gene polymorphisms in interleukin-10 and transforming growth factor beta 1 in sarcoidosis. AB - BACKGROUND AND AIM OF WORK: Interleukin-10 (IL-10) and transforming growth factor beta 1 (TGF-beta 1) are anti-inflammatory cytokines that play important roles in the immunoregulatory processes of numerous granulomatous diseases. In sarcoidosis polymorphisms (PMs) within these cytokine genes are suspected of modifing the course of the disorder. Therefore, we were interested in whether the genotype frequencies for a PM at position -1082 of the IL-10 or in codon 25 of the TGF beta 1 gene differ in sarcoidosis or its distinct phenotypes in comparison with healthy individuals. METHODS: In 51 sarcoidosis patients and 72 healthy blood donors, genotyping for the named PMs was performed by PCR methodology and restriction enzyme digestion. Patients were retrospectively classified according to their course of disease, namely spontaneous remission, regressive under therapy, or chronic-progressive. RESULTS: For TGF-beta 1 PM the genotype frequencies ranged between 81.8-90.5, 9.6-13.9 and 0-5.3 percent for genotype GG, CG and CC respectively. For IL-10 PM the values ranged between 17.7-23.2, 54.4 68.4 and 21.1-26.4 percent for AA, AG and GG. Statistical comparisons of the allele and genotype frequencies between the clinical defined sarcoidosis groups and the healthy blood donors revealed no significant differences. PMID- 11436537 TI - Relationship between outcome measures of six-minute walk test and baseline lung function in patients with interstitial lung disease. AB - BACKGROUND AND AIM OF THE WORK: In patients with interstitial lung disease (ILD), the six-minute walk test (6MWT) has been rarely used, and up till now, the relationship between outcome measures of the test and baseline lung function has not yet been examined. Therefore, we assessed walk distance, oxygen desaturation, and breathlessness perception during 6MWT, and their relationships to baseline lung function in patients with ILD. METHODS: Forty ILD patients with history of breathlessness during physical exertion performed a 6MWT following a standard protocol. Breathlessness perception during walk was assessed by visual analogue scale (VAS, in mm). RESULTS: The mean walk distance was 487 meters (range 271 689). Mean baseline oxygen saturation (Base SpO2, %) was 94% and was reduced during walk, either as mean oxygen saturation (Mean SpO2, 89%, p < 0.001) or as mean fall in oxygen saturation during walk (Fall SpO2, 5%). Furthermore, VAS significantly increased after walk (5 mm to 44 mm, p < 0.001). A close relationship of TLco and TLC to walk distance and Fall SpO2 was found (r = 0.45 and 0.42 and r = -0.75 and -0.64, respectively; p < 0.001, each). On the basis of regression equations by stepwise multiple regression analysis, walk distance was predicted by age and FVC (r2 = 0.50), Mean SpO2 by TLco and Base SpO2 (r2 = 0.80), and Fall SpO2 only by TLco (r2 = 0.57). Breathlessness perception was not reliably predicted by any assessed variables. CONCLUSIONS: We confirmed that 6MWT provides a global evaluation of sub-maximal exercise capacity in ILD patients. We also found that walk distance and oxygen desaturation, but not breathlessness perception during walk, are strictly related to baseline lung function. PMID- 11436538 TI - Sarcoidosis in Korea: report of the Second Nationwide Survey. AB - BACKGROUND: In Korea, sarcoidosis is a very rare disease but the first nationwide survey performed in 1992 by the Korean Academy of Tuberculosis and Respiratory Diseases revealed 113 biopsy-proven cases. To investigate whether sarcoidosis is continuously increasing in Korea and verify the clinical pattern, the second nationwide survey was performed. METHOD: The questionnaires were sent to hospitals of larger than 80-bed size for collection of biopsy-proven sarcoidosis patients newly diagnosed between July 1992 and June 1999. RESULT: A total of 309 biopsy-proven cases was recruited from 58 hospitals. The incidence has gradually increased from 14 cases in 1993 (0.027/100,000) to 59 cases in 1998 (0.125/100,000). The peak age was in the thirties (33%) and females were predominant (64.6%). Not a single case was detected among 190,029 mass chest X ray films taken in the national tuberculosis surveys from 1980 to 1995. The most common symptom was respiratory (42%) and the most frequently involved organ was the lung (87%), followed by skin (31%). Tuberculin skin anergy was found in 82.2%. Hypercalcemia was found in 9.7% and hypercalciuria in 22.5%. FVC and FEV1 were reduced in about a quarter of the patients and reduced DLCO was found in 37%. CONCLUSION: Sarcoidosis is still very rare in Korea even though it is increasing slowly and clinical manifestation was similar to the western pattern. PMID- 11436539 TI - Fungal infections in lung transplantation. Incidence, risk factors and prognostic significance. AB - BACKGROUND AND AIM OF THE WORK: Fungal infections are frequent following lung transplantation and are associated with high mortality and morbidity. The study aims at 1) reporting our experience with fungal infections after lung transplantation; 2) identifying statistically significant correlations between the occurrence of fungal infections and bacterial infections, cytomegalovirus disease, rejection and steroid therapy; 3) assessing whether the presence of fungal infection has an impact on long-term survival. METHODS: 60 lung transplant recipients were studied with respect to incidence, pattern of presentation and median time to presentation of fungal infection after the transplant. Correlation analysis of the variables of interest was undertaken in 30 patients who had a minimum follow-up of 1 year following transplant. RESULTS: The prevalence of fungal infection was 44%; severe infections occurred in 5 patients (11%). The presence of Candida preoperatively was not associated with an increased risk of fungal infection. In a logistic regression analysis, a significant correlation was found between the occurrence of fungal infection and the following variables: respiratory bacterial infections (p = 0.0003), cytomegalovirus disease (p = 0.00001) and steroid therapy (p = 0.04). No statistically significant difference was found between patients who experienced a fungal infection and those who did not, either in univariate or multivariate survival analysis (p = 0.08). CONCLUSIONS: 1) fungal infections are frequent in lung transplant recipients and may be severe in more than 10% of the cases; 2) the presence of fungi preoperatively is not a contraindication to transplantation: an antifungal prophylaxis is probably indicated in such cases postoperatively; we recommend the use of the less nephrotoxic liposomal Amphotericin B by aerosol route; 3) a statistically significant association exists between fungal infections and both steroid therapy and CMV disease; this suggests that a similar antifungal prophylaxis is indicated in these clinical circumstances; 4) the presence of fungal infection is not an independent prognostic factor of long-term survival. PMID- 11436540 TI - The value of labial biopsy in the differentiation of sarcoidosis from tuberculosis. AB - BACKGROUND: Since the clinical features of sarcoidosis and tuberculosis may mimic each other, and that differentiation is not easy on clinical grounds, a histologic diagnosis may be mandatory in countries where the prevalence of tuberculosis is high or in populations with large numbers of immigrants from those countries. Previous studies have suggested the minor salivary gland biopsy as a useful method in the diagnosis of sarcoidosis. The present study was undertaken to evaluate the value of labial biopsy in the differentiation of sarcoidosis from tuberculosis in patients with enlarged hilar and paratracheal lymph nodes. METHODS: Labial biopsy was performed in 50 consecutive patients with sarcoidosis, and in 35 consecutive patients with tuberculosis who had intrathoracic lympadenopathy. The files of all patients were reviewed for the clinical presentation, radiographic features, SACE levels, tuberculin skin test anergy, and the frequency of positive labial biopsy in each disease. RESULTS: Noncaseating granulomas were present in labial biopsies obtained from 24 patients (48%) of 50 patients with sarcoidosis. Labial biopsies were positive in 4 of 6 patients who had an abnormality on eye examination and in 3 of 5 patients who had noncaseating granulomas on biopsy material from skin. In two of 4 patients who underwent mediastinoscopy, noncaseating granulomas were detected on labial biopsy. In contrast to the patients with sarcoidosis labial biopsies revealed normal minor salivary glands in all patients with tuberculosis. CONCLUSIONS: Labial biopsy has a high discriminatory value as a diagnostic tool in the differentiation of sarcoidosis from tuberculosis. Although it has a rather lower diagnostic yield than transbronchial lung biopsy, labial biopsy should be considered as a first line approach prior to performing other more invasive procedures for the tissue confirmation of sarcoidosis. PMID- 11436541 TI - Long-lasting myopathy as a major clinical feature of sarcoidosis in a child: case report with a 7-year follow-up. AB - Muscle involvement in sarcoidosis is rarely described as the predominant feature and muscular symptoms are seldom observed. In recent pediatric series, sarcoid myopathy was no longer considered a typical aspect of sarcoidosis. The authors report a case of sarcoidosis in a patient presenting predominant muscular symptoms since childhood, due to biopsy-proven muscle localization. A seven-year follow-up has demonstrated a slow improvement of symptoms with persistency of electromyography (EMG) and biochemical abnormalities. Mild and transient pulmonary involvement was demonstrated only after diagnosis. Clinical improvement associated with a decrease in serum muscular enzyme levels with no changes in EMG was observed after a six-month course of systemic corticosteroid therapy. In childhood, skeletal muscle symptoms may be the presenting feature of sarcoidosis. PMID- 11436542 TI - Deep venous thrombosis as a possible complication of interferon-gamma treatment. PMID- 11436543 TI - Differences in functional immune responses of high vs. low hardy healthy individuals. AB - An association between the personality trait of hardiness and immune responses was explored. Blood samples were collected from 21 healthy individuals under nonstressful conditions, who had either high or low levels of hardiness. Functional immune assays tested for natural killer (NK) cell activity and proliferation responses to Candida albicans (Candida), purified protein derivative from Mycobacterium tuberculosis (PPD), lipopolysaccharide (LPS), concanavalin A (Con A), and Staphylococcus enterotoxin A (Staph A). Differences between high and low hardy groups on these immune responses were examined using Bonferroni adjusted independent t-tests. Results revealed significant differences between the groups for Candida, PPD, Con A, and Staph A. In all instances, the high hardy group had larger mean proliferative responses than the low hardy group. Implications of the study as well as avenues for future research are discussed. PMID- 11436544 TI - Social support, intrusive thoughts, and quality of life in breast cancer survivors. AB - This study explores the moderating effect of social support on the relationship between cancer-related intrusive thoughts and quality of life. Sixty-four breast cancer survivors completed self-report measures of appraisal social support (the disclosure of thoughts and feelings to significant others), cancer-related intrusive thoughts, and quality of life. Controlling for demographic and treatment variables, the negative impact of cancer-related intrusive thoughts on both physical and mental quality of life measures was moderated by appraisal social support. For women with high levels of appraisal support, cancer-related intrusive thoughts had no significant relationship with quality of life. However, for women with low levels of appraisal support, the relationship between cancer related intrusive thoughts and quality of life was significant and negative. These results suggest that appraisal social support can mitigate the impact of traumatic life events. PMID- 11436545 TI - Psychological and screening profiles of first-degree relatives of prostate cancer patients. AB - The purpose of the present study was to systematically compare the psychological and screening profiles of first-degree relatives (FDRs) of prostate cancer patients versus non-FDRs. FDRs (n = 56) and non-FDRs (n = 100), recruited through prostate cancer index cases and newspaper advertisements, completed questionnaires via mail. FDRs reported feeling at greater risk for prostate cancer, estimated that they were at higher average lifetime risk for the disease, agreed more strongly that prostate cancer is inherited, and that less can be done to prevent the development of the disease. Increased age, but not FDR status, was associated with more frequent screening behavior. Taken together, the results indicate that FDRs are characterized by greater perceived vulnerability to prostate cancer and lower expectations about disease prevention. Yet, they are no more likely to be screened than non-FDRs. These findings underscore the importance of developing, and evaluating, evidence-based health communication protocols to promote screening adherence among at-risk patients. PMID- 11436546 TI - The role of illness, resources, appraisal, and coping strategies in adjustment to HIV/AIDS: the direct and buffering effects. AB - This study examined the utility of a stress and coping model of adjustment to HIV/AIDS. A total of 114 HIV-infected gay or bisexual men were interviewed and they completed self-administered scales. Predictors included illness variables (disease stage and number of symptoms), coping resources (optimism and social support), appraisal (threat, challenge, and controllability), and coping strategies (problem- and emotion-focused). Adjustment outcomes were depression, global distress, social adjustment, and subjective health status. Results from hierarchical regression analyses indicated that better adjustment was related to an asymptomatic illness stage, fewer HIV-related symptoms, greater social support, challenge and controllability appraisals, problem-focused coping, and lower threat appraisals and reliance on emotion-focused coping. There was limited support for the stress-buffering effects of optimism. Findings support the utility of a stress and coping model of adjustment to HIV/AIDS. PMID- 11436547 TI - Behavioral treatment of insomnia: treatment outcome and the relevance of medical and psychiatric morbidity. AB - Recently, we undertook a case series study and found that behavior therapy for insomnia was effective as plied in the clinic setting and that the findings were similar to those in the "clinical trial" literature. In the present study, we evaluate a second set of case series data to assess (1) the replicability of our original findings, (2) if our treatment outcomes are statistically comparable to those in the literature, and (3) if medical and psychiatric morbidity influence treatment outcome. It was found that patients who completed four or more sessions of cognitive behavioral therapy for insomnia (CBT) were, on average, 33% improved. This average corresponded to a 56% reduction in wake time after sleep onset, a 34% reduction in sleep latency, a 29% increase in total sleep time, and a 13% decrease in number of awakenings per night. These findings are not significantly different from those reported in literature for both CBT and pharmacotherapy interventions. Medical and psychiatric comorbidity did not influence treatment outcome. PMID- 11436548 TI - [Interspecific crosses in Avena L. species]. AB - Problems of crosses between various oat species are considered with regard to establishing their taxonomic positions and genomic compositions of individual species. The evolution of the genus and approaches to the search for the diploid and tetraploid ancestor of the hexaploid species are considered. Use of wild oat species in breeding is demonstrated. The results of studies of the gene pool of wild oat species are presented. These studies were performed at the Vavilov All Russian Plant Breeding Institute with the purpose of solving problems of phylogeny and practical breeding. PMID- 11436549 TI - [Homologous recombination and chromosomal rearrangements in Escherichia coli strains carrying a heterozygous tandem duplication]. AB - Heterozygous tandem duplications formed in conjugational matings in Escherichia coli provides a convenient model system for studying the evolution of bacterial chromosome. Heterozygous duplications segregate various classes of haploid and diploid recombinants that appear as a result of unequal crossing over between sister chromosomes. In this work, an extended tandem duplication in the deo operon of E. coli carrying deoA deoB::Tn5/deoC deoD thr::Tn9 alleles was examined. Recombination between homologous DNA repeats in the duplication was studied in strains carrying different combinations of recBC, sbcBC, recB::Tn10, recQ::Tn3 mutations. The frequency of recombination between homologous DNA repeats was very high in all strains and did not decrease when the RecBCD and RecF recombinational pathways were simultaneously damaged in strains with the recB sbcBC recQ (or recF) genotype. It is assumed that unequal crossing over between direct DNA repeats in duplications may proceed through a particular pathway of "adaptive" recombination. PMID- 11436550 TI - [Suppression of frameshift mutation as a result of partial inactivation of translation termination factors in Saccharomyces cerevisiae yeast]. AB - Special search for frameshift mutations, which are suppressed by the cytoplasmic [PSI] factor and by omnipotent nonsense suppressors (recessive mutations in the SUP35 and SUP45 genes), partially inactivating a translation termination complex, was initiated in the LYS2 gene in the yeast Saccharomyces cerevisiae. Mutations were obtained after exposure to UV light and treatment with a mixture consisting of 1.6- and 1.8-dinitropyrene (DNP). This mixture was shown to induce mutations of the frameshift type with a high frequency. The majority of these mutations were insertions of one A or T, which is in good agreement with the data obtained in studies of DNP-induced mutagenesis in other eukaryotes. Frameshift suppression in yeast was first shown on the example of the mutation obtained in this work (lys2-90), which carried the insertion of an extra T in the sequence of five T. This frameshift suppression was shown to occur in the presence of the [PSI] factor (i.e., due to the prion form of the translation release factor eRF3) and as a result of mutations in genes SUP35 or SUP45, which partially inactivate translation termination factors eRF3 and eRF1, respectively. Alternative mechanisms of programmed translational frameshifting in the course of translation and the possibility of enhancing the effectiveness of such frameshifting in the presence of the [PSI] factor are considered. PMID- 11436551 TI - [Genetic diversity of a natural population of Sinorhizobium meliloti, detected during analysis of a cryptic plasmid and ISRm2011-2 fingerprints]. AB - Fifty-six natural strains of alfalfa nodule bacteria were isolated from samples of the soil under wild legume and alfalfa in two different field sites of Irkutsk oblast. Based on the results of analysis of plasmid profile, 11 different types of strains were detected, and 43 types were identified based on the results of hybridization with the insertion sequence element ISRm2011-2. Significant differences were found in the plasmid profile and IS fingerprints between strains isolated from the soil under alfalfa and the soil under legume. In contrast, strains growing at some distance from each other differed only in the IS fingerprints. From a comparison of results obtained in the assessment of plasmid profile and in analysis of IS fingerprints with results of RFLP analysis in strains, the conclusion about the transference of cryptic plasmids between strains and genetic rearrangements in strains of this population was drawn. PMID- 11436552 TI - [Mutagenic potential of copper compounds and modification of effects of silver iodide]. AB - Mutagenic potential of copper compounds and its alteration in case of the interaction with silver compounds were analyzed by use of plant test systems. As test systems, Crepis capillaris L., Tradescantia clone 02, and soybean (Glycine max (L.) Merrill) were used. Mutagenic properties of copper iodide and copper sulfate were not detected. CuI, being not a mutagen by itself, remarkably enhanced mutagenic potential of AgI. PMID- 11436553 TI - [Effect of rye chromosomes on features of androgenesis in wheat-rye substituted lines of Triticum aestivum L. sort Saratovskaya 29/Secale cerale L. sort Onokhoiskaia and Triticale]. AB - The characteristic features of androgenesis in six wheat-rye substitution lines Triticum aestivum L. (cv. Saratovskaya 29)/Secale cereale L. (cv. Onokhoiskaya) and triticale (2n = 56) using anther culture at different concentrations of 2,4-D in the growth medium were studied. Under variable cultivation conditions, the significant effect of genotypic diversity on the variability of such androgenesis parameters as the frequency of productive anthers, the frequency of embryoid formation, and the frequency of total regenerated plantlets, was shown. It was demonstrated that chromosomes 1R, 3R, and 7R stimulated the formation of androgenous embryoids, while chromosome 5R produced an opposite effect. In triticale and substitution lines, the regeneration ability of androgenous embryoids induced by elevated 2,4-D concentrations was inhibited. Chromosome 1R of the Onokhoiskaya cultivar was suggested to contain genes suppressing regeneration of green plantlets, while chromosome 3R, conversely, stimulated their formation. Chromosomes 1R, 2R, 3R, and 7R of the Onokhoiskaya cultivar did not inhibit the spontaneous formation of androgenous hexaploids in the substitution lines. PMID- 11436554 TI - [Characteristics of some breeds of swine and populations of Georgian and Western Siberian wild boar by immunogenetic systems of blood serum proteins]. AB - A bank of reagents for hog serum protein allotypes has been created. All of these allotypes passed international comparison tests in 1987-1988. The bank can be used for typing the animals for four generally accepted (Gp, LpB, Lpr, and IgGH) and several experimental systems. In this study, immunogenetic characteristics of some pig breeds (Large White, Lithuanian White, Swedish Landrace, Kakhetinskaya, and Svanetskaya) bred in Georgia are compared with those of western Siberian breeds (Large White, Kemerovskaya, and Northern Siberian), and some foreign breeds, as well as with European, Caucasian, and Siberian subspecies of the wild boar. PMID- 11436555 TI - [Effect of four mutations (Cr,S,S(H),h) in genes for mink coat color on brain monoamine oxidase]. AB - Activity of A and B types of monoamine oxidase (MAO) has been investigated in the brain stem and brain hemispheres of mink males of five genotypes for coat-color mutations: standard dark-brown (+/+); heterozygous for the semidominant mutation Black crystal (Cr/+); homozygous for the semidominant mutation Black cross, or "95% White" (S/S); heterozygous for the semidominant mutation Shadow (SH/+); and homozygous for the semirecessive mutation hedlum white (h/h). The main changes in the activity of the A and B MAO types occur in the brain hemispheres. A reduced activity of MAO A has been recorded in the hemispheres of Black crystal minks (Cr/+) and an elevated activity, in the hemispheres of Shadow (SH/+) and 95% White (S/S). The activity of MAO B is reduced in the hemispheres of Black crystal and elevated in the hemispheres of hedlum white (h/h). An increased MAO A activity has also been recorded in the brain stem of Shadow minks (SH/+). It is suggested that genes controlling coat color have a pleiotropic effect on sexual behavior in males and the endocrine function of testicles mediated by a putative change in the metabolism of brain neurotransmitters, substrates of MAOs A and B. PMID- 11436556 TI - [Expression of the proopiomelanocortin gene in the hypophysis of NICAG-line hypertensive rats]. AB - Expression of the proopiomelanocortin (POMC) gene in NISAG rats with inherited stress-induced arterial hypertension has been investigated by the reverse transcription-competitive PCR method. Normotensive rats of the inbred Wistar albino Glaxo (WAG) strain have been used as control. The effect of 3 h immobilization stress has been examined. The content of POMC mRNA in pituitaries of stressed NISAG rats exceeds that in non-stressed rats of the same strain by a factor of 2.4. Rats of the WAG strain do not demonstrate a significant increase in the level of POMC mRNA. In addition, the basIC transcription level of the POMC gene in NISAG rats is higher than in WAG ones. PMID- 11436557 TI - [Effect of genotype and social stress on cAMP- and substrate-dependent mechanisms of regulating hormonal function of testis in mice]. AB - Several steps of cAMP- and substrate-dependent testosterone production in the testes were studied with laboratory mouse micropopulations of six inbred strains (A/He, CBA/Lac, C57BL/6J, DD, YT, PT). The strains differed in basal testosterone production in the gonads and in its response to activation of the adenylate cyclase signal transduction pathway at various steps by human chorionic gonadotropin (hCG), the cholera toxin, forskolin, and dibutyryl-cAMP and in the presence of pregnenolone, an early precursor of testosterone. Establishment of dominant-subordinate relationships in mouse populations substantially affected testosterone production in response to all activators of testicular steroidogenesis. The secretory activity of the testes decreased at the early establishment of social hierarchy in experimental micropopulations, then returned to the initial level, and again decreased in the case of activation with hCG, dibutyryl-cAMP, and pregnenolone. With all activators of steroidogenesis, basal and activated testosterone production changed in the same direction during the establishment and maintenance of social hierarchy, suggesting coordinated changes in all examined steps of testosterone biosynthesis in the testes. The among strain differences in response to all activators of steroidogenesis remained much the same at various stages of the establishment of social hierarchy. The parameters of cAMP- and substrate-dependent testosterone production averaged over individual stages of the establishment of social hierarchy proved associated. Their genotypic correlations were positive and, in many cases, significant. Subsequent component analysis showed that one principal component accounted for more than 80% of the total among-strain variation, suggesting a coordinated genetic control of the endocrine function of the testes. PMID- 11436558 TI - [Determination of the minimum size of Gallus gallus domesticus chicken microchromosome by a pulse electrophoresis method]. AB - The karyotype of the chicken Gallus gallus domesticus was studied by means of pulsed-field electrophoresis. An electrokaryogram was obtained for the microchromosomal (MI) portion of the chicken genome. Chicken MIs were separated into two fractions. A fraction with a higher mobility included MIs sized 3.4-4.8 Mb; the lower size limit of a less mobile fraction corresponded to MIs of approximately 5 Mb. The smallest MI in the chicken karyotype was estimated at 3.4 +/- 0.25 Mb. PMID- 11436559 TI - [Molecular-genetic analysis of polymorphism of the DXS532 locus in people from the Volga-Ural region]. AB - The DXS52 polymorphic locus mapping to the 5'-region of the blood-clotting factor VIII gene on the X chromosome was genotyped in seven Volga-Ural ethnic groups (Bashkirs, Tatars, Chuvashes, Maris, Mordovians, Udmurts, and Komis). A total of 47 different genotypes and 15 allelic variants of this locus were described. Substantial intra- and interpopulation heterogeneity of the ethnic groups studied in respect to frequency and distribution of the DXS52 alleles and genotypes was demonstrated. The unimodal DXS52 allele frequency distribution pattern with the peak at 1690 bp was typical to Mordovians and Komis. Chuvashes and Maris, as well as Udmurts, were characterized by bimodal frequency distribution patterns, with the peaks at 1690 and 670 bp, and 1690 and 1390 bp, respectively. Moreover, Bashkirs and Tatars displayed trimodal DXS52 allele frequency distribution patterns with the peaks at 1690, 1390, and 670 bp. The DXS52 allele frequency distribution patterns described in populations of the Volga-Ural region were found to be remarkably different from those established for the mixed Moscow population and the population of Western Europe. These data indicate that the DXS52 locus is highly informative, and this polymorphic system can serve as a molecular marker for population genetic studies. PMID- 11436560 TI - [Endothelial nitric oxide synthase gene minisatellite polymorphism: study in populations of the Volga-Ural region and analysis of associations with myocardial infarct and essential hypertension]. AB - The 27-bp tandem repeat polymorphism in intron 4 of the endothelial nitric oxide synthase gene (eNOS) in populations of the Volga-Ural region was studied by means of polymerase chain reaction. In Russians and Tatars, the possible association of this polymorphism with coronary heart disease complicated by either myocaridal infarction or by essential hypertension was examined. Russians with essential hypertension associated with hypertrophy of the left ventricle displayed a statistically significant increase of the eNOS4A/B genotype and the A allele frequencies along with the decrease of the frequencies of the eNOS4B/B genotype and the B allele. In Tatars survived from myocardial infarction and with the risk of cardiovascular diseases (smoking or burdened heredity), a statistically significant increase of the frequencies of the eNOS4A/B genotype and the A allele was observed. Thus, in Russians the eNOS4A/B genotype was associated with the development of essential hypertension, while in Tatars it was associated with the risk of myocardial infarction. PMID- 11436561 TI - [Effect of the frequency of X-monosomal cell clone on variability of anthropometric indicators in Shereshevsky-Turner syndrome]. AB - Using methods of mathematical statistics the relationships were determined between 31 anthropometric traits (ATs) and the frequency of the X-monosome cell clone in 53 patients with either 45, X-monosomy or mosaic forms (45,X/46,XX) of the Shereshevsky-Turner syndrome (STS). AT variations were studied in patients untreated with growth hormone and in 25 control fertile healthy women. In 29 patients, the degree of mosaicism was assessed by interphase FISH analysis using X-centromer-specific DNA probe hybridized to the cell nuclei of two types of tissues differing in embryonic origin (lymphocytes and oral epithelium, originating from meso- and ectoderm, respectively). The level of X-monosome mosaicism had a substantial effect on some AT, which depended similarly on the proportion of X-monosome cells in tissues of different embryonic origin. Statistically significant negative correlations were revealed between the size of X-monosome clone and 13 height-weight, longitudinal, and circumference traits, whereas positive correlations were characteristic of seven mostly width traits. Eleven ATs showed no correlation with the X-monosome cell clone. Discriminant analysis of all ATs, whose variations depended on the frequency of X-monosome cell clone, was found to be an essential tool for precise classification of both STS patients with different degree of mosaicism and healthy women. Based on these results, the set of ATs characteristic of the STS phenotype was identified. PMID- 11436562 TI - [Malecot's parameters of isolation by distance and the degrees of endogamy in three regions of the Chuvash Republic]. AB - Malecot's parameters of isolation by distance and the degree of endogamy were estimated for individual Village Soviets of the Kanash, Cheboksary, and Morgaushi raions of Chuvashiya. Inbreeding landscape schemes were constructed for the Kanash and Morgaushi raions. The suitability of the Malecot's model of isolation by distance for populations of the Village-Soviet level was analyzed. PMID- 11436563 TI - [Three new mutations in the porphobilinogen deaminase gene, detected in acute intermittent porphyria patients from Russia]. AB - Porphobilinogen deaminase (PBGD) is a key enzyme of the heme biosynthetic pathway. Defects in the PBGD gene lead to an autosomal dominant disease, acute intermittent porphyria (AIP). Almost all AIP patients with rare exceptions are heterozygous for the defective gene. To date, at least 160 different mutations causing AIP are identified. Extensive investigations along this line are conducted in many countries of the world. In Russia these studies had not been hitherto performed. Here we report the results of molecular genetic examination of four Russian patients with AIP diagnosed from clinical symptoms. By direct sequencing of the PBGD gene or the corresponding cDNA, we have detected four mutations, three of which were not previously encountered in the world population. These are TAAG deletion in intron 7 between positions +2 and (IVS7 2 5 delTAAG); T deletion in the initiation codon ATG of exon 3, and the G for C replacement at position -1 of intron 5 (IVS5 as -1 G:C), which disrupts splicing. In addition, in one female patient, a known deletion CT in codon 68 was revealed. In two patients, expression of PBGD gene alleles was significantly disproportional, so that normal mRNA prevailed in one case and mRNA of nonerythroid type in the other. Deletion in intron 7 was easily detectable due to the formation of a heteroduplex fragment with abnormal electrophoretic mobility directly in PCR. This simple heteroduplex analysis allowed us to exclude AIP carriage in son and daughter of a female patient with the genetic defect. PMID- 11436564 TI - [Genetic predisposition to development of toxic liver cirrhosis caused by alcohol]. AB - Comprehensive analysis of the contribution of genetic factors into predisposition to alcoholic toxic cirrhosis (TC) was performed. The ABO, RH, HP, TF, GC, PI, ACP1, PGM1, ESD, GLO1, and GST1 genetic polymorphisms were compared in 34- to 59 year-old male TC patients and control donors of the same sex and age. The phenotypic frequencies in the TC group deviated from the theoretically expected values; the main difference was the excess of rare homozygotes for the loci GC, ACP1, ESD, and GLO1. In the TC patients, the observed heterozygosity (Ho) was considerably lower than the theoretically expected value (H(e)). Wright's fixation index (F) in the TC patients was 30 times higher than in the control group (0.0888 and 0.0027, respectively). The frequencies of PI*Z and PI*S, the PI alleles that are responsible for lower concentrations of proteinase inhibitor, were 12 and 6 times higher in the TC than in the control group. The TC patients exhibited a significantly higher frequency of the liver glutathione-S-transferase GST1*0 allele, whereas the GST1*2 frequency was two times higher in the control subjects than in the TC patients (0.2522 and 0.0953, respectively). The TC and control groups showed statistically significant differences in the frequencies of the following alleles of six independent loci: ABO*0, TF*C1, TF*C2, PI*M1, PI*Z, ACP1*C, PGM1*1+, PGM1*1-, PGM1*2-, GST1*0, and GST1*2. The haptoglobin level was significantly higher and the serum transferrin level was drastically lower in all phenotypic groups of TC patients than in control subjects. The concentrations of IgM and IgG depended on the HP, GC, and PI phenotypes. The total and direct reacting bilirubin concentrations depended on the erythrocytic-enzyme phenotypes (ACP1, PGM1, and GLO1) in both TC and control groups. PMID- 11436565 TI - [Modification of a method for detecting the post-phoretic activity of mannose (Mpi, EC 5.3.1.8)- and glucose (Gpi, EC5.3.1.9)-6-phosphate isomerase]. AB - A method for detecting activities of mannose- and glucose-6-phosphate isomerases based on enzyme production of the substrates is described. The results obtained for several animal taxa are illustrated by photographs. PMID- 11436566 TI - [Polymorphic markers for the angiotensinogen and angiotensin-converting enzyme in Yakuts. Lack of association with blood pressure level]. AB - Allele frequency distributions of the insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme (ACE) gene and the M235T polymorphism of the angiotensinogen gene was studied in a random sample of the indigenous population of the Sakha Republic. The allelic variants of these genes did not showed an association with blood pressure in Yakuts. PMID- 11436567 TI - Perspectives. Rural care not on life support, but PPS changes needed. PMID- 11436568 TI - CT evaluation of the liver: principles and techniques. AB - Technical advances within the past decade have dramatically improved the diagnostic capabilities of CT, primarily due to helical CT scanners coupled with mechanical injectors for i.v. administration of contrast media. There is no such thing as a generic "abdominal CT scan"; rather, specific protocols should be utilized for optimal detection and characterization of hepatic masses (or other suspected pathologic processes). Multiphasic CT protocols are especially important for detecting hypervascular liver tumors. We present the principles and rationale for various CT protocols along with some of the common pitfalls. Understanding this background material will allow the reader to better understand subsequent articles dealing with specific applications of CT and other abdominal imaging techniques. PMID- 11436569 TI - Liver mass evaluation with ultrasound: the impact of microbubble contrast agents and pulse inversion imaging. AB - Liver mass evaluation includes two essential elements--lesion detection and lesion characterization. Both of these are greatly improved on sonography with the addition of contrast agents and the use of specialized imaging techniques, particularly pulse inversion imaging. Ultrasound contrast agents are comprised of tiny microbubbles of gas that interact with the ultrasound beam producing an enhancement of the Doppler signal from blood. Pulse inversion imaging allows preferential detection of the signal from the microbubble agents with suppression of the signal from background tissue. Two imaging techniques include a low mechanical index (MI) nondestructive method to show lesional vascularity and a high MI destructive mode that produces disruption of the bubbles in a single frame. The latter allows for quantitative assessment of the relative enhancement of a lesion as compared with the adjacent liver parenchyma, which is a reflection of the relative vascular volumes. Vascular imaging has shown characteristic and reproducible features of common liver masses, including hemangioma, focal nodular hyperplasia, hepatocellular carcinoma, and liver metastases. Delayed postvascular enhancement of the normal liver, a phenomenon that is unique to certain classes of microbubble contrast agents, allows detection of more and smaller malignant lesions than on baseline. PMID- 11436570 TI - MRI evaluation of the liver. AB - Following the introduction of rapid, high-quality scan techniques and the development of new, tissue-specific contrast agents, the applications of magnetic resonance imaging (MRI) for liver imaging continue to grow. MRI using a T1 weighted sequence, T2-weighted sequence, and serial gadolinium-enhanced gradient echo sequences is very effective at both detecting and characterizing various hepatic lesions. The use of a growing number of contrast agents has the potential to increase the sensitivity and specificity of liver MRI in numerous pathological conditions by improving morphological and functional information. This article examines the current status of liver MRI, highlighting technical and methodological approaches, contrast agents, and main clinical applications. MRI appears to be an ideal diagnostic tool for the detection and characterization of liver disease. PMID- 11436571 TI - Radionuclide imaging of hepatic and biliary disease. AB - This article will focus on common clinical applications of scintigraphy in focal hepatic lesions, acute cholecystitis, biliary dyskinesia, biliary obstruction, postoperative liver and biliary tract, and neonatal cholestasis. The utility of positron emission tomography will also be addressed. PMID- 11436572 TI - Imaging of diffuse liver disease. AB - Advances in imaging technology and development of liver-specific contrast agents have significantly increased the role of radiology in the detection and characterization of processes diffusely involving the liver. Tailored magnetic resonance imaging (MRI) sequences allow an accurate detection of many storage and metabolic diseases, such as iron overload disorders and steatosis (fatty liver). Faster scanning techniques available with both computed tomography (CT) and MRI provide, by assessing contrast dynamics, sufficient information for the characterization of diffuse neoplastic and vascular disorders. Characteristic changes in attenuation on CT, signal intensity on MRI, and enhancing features can be used to diagnose specific diffuse diseases such as candidiasis, diffuse/multifocal hepatocellular carcinoma, and schistosomiasis. Although an overlap in imaging findings still exists, familiarity with the imaging features of uncommon disorders such as Wilson's disease, amyloidosis, and sarcoidosis may be diagnostic in the proper clinical setting. This review focuses on the current role of imaging in the detection and characterization of diffuse liver disorders. Recent developments that have amplified the role of noninvasive diagnostic evaluation of these conditions are especially highlighted. PMID- 11436573 TI - Imaging evaluation of the cirrhotic liver. AB - Because recent advances in medical care decrease the mortality rate due to liver cirrhosis itself, many cirrhotic patients die due to hepatocellular carcinoma. Accordingly, the role of radiology in the evaluation of the patient with cirrhosis is primarily to characterize the morphologic manifestations of the disease, evaluate the hepatic and extrahepatic vasculature, assess the effects of portal hypertension, and detect hepatic tumors. When the latter are identified, a critical role of imaging technology is to differentiate hepatocellular carcinoma from other nodular lesions, such as dysplastic nodules and regenerating nodules. Screening strategies for patients with cirrhosis have been proposed to facilitate the detection of small, asymptomatic hepatocellular carcinomas. Dynamic studies using computed tomography (CT) and magnetic resonance imaging (MRI) are very useful for the diagnosis of hepatic tumors previously detected by ultrasound, as well as for screening. In Japan, patients with documented cirrhosis typically undergo serum alpha-fetoprotein testing and/or PIVKA-II (protein induced by vitamin K absence or antagonist II) measurements every 2 months, ultrasound every 3 months, and CT or MRI every 6 months. This has resulted in great success in detecting small hepatocellular carcinomas (less than 2 cm in diameter) and early stage well-differentiated hepatocellular carcinomas. PMID- 11436574 TI - Evaluation of the liver for metastatic disease. AB - Metastatic disease of the liver accounts for the vast majority of detected liver masses. In patients with suspected metastatic disease, cross-sectional imaging with ultrasound (US), computed tomography (CT), or magnetic resonance imaging (MRI) is critical. In the group of patients undergoing evaluation for hepatic surgery, it is even more important to optimize techniques to detect and localize metastatic disease. With improvements in technology and contrast agents, there are several approaches to imaging the liver for metastatic disease. The approach will vary by institution. This article will attempt to provide an overview of the general issues relevant to imaging metastatic disease, highlight the advantages and disadvantages of one modality compared to another, and illustrate the appearance of metastases using US, CT, and MRI. PMID- 11436575 TI - Imaging of benign hepatic masses. AB - Using helical CT and rapid i.v. injection of contrast material, benign hepatic masses are discovered in at least 20% of subjects. To minimize expensive and invasive evaluation, an orderly approach to the interpretation of these lesions is necessary. Many benign lesions have a near pathognomonic appearance on a properly performed and interpreted CT scan. Others can have the diagnosis confirmed by a specific and well-chosen follow-up study, such as contrast enhanced MR. In this article we present the most common types of benign hepatic masses and our approach to their diagnosis. PMID- 11436576 TI - Pediatric liver imaging. AB - The evaluation of hepatic diseases in children is often a multimodality process, requiring multiple imaging tests to determine the cause and extent of an abnormality. Ultrasound (US), computed tomography (CT), and magnetic resonance imaging (MRI) have distinct roles to play in the evaluation of hepatic disease in children. This article addresses the hepatic and biliary lesions that are unique or more common in children and describes their cross-sectional imaging characteristics. In addition, the techniques and protocols for US, CT, and MRI are reviewed. PMID- 11436577 TI - Imaging for right lobe living donor liver transplantation. AB - The authors describe their experience using multidetector, multiphase computed tomography (CT) for the preoperative evaluation of donors undergoing adult right lobe liver transplantation. The marked improvement in image resolution has been extremely valuable in delineating small hepatic arterial structures, including accessory and replaced vessels. Assessment of liver parenchyma is valuable in determining the presence of steatosis or other unexpected abnormalities. Measurement of total liver volume is highly accurate and reproducible. Virtual hepatectomy can be performed in an avascular plane between the right and left lobes, to the right of the middle hepatic vein. Knowledge of the resultant right and left lobe volumes is critical in patient selection. Multidetector, multiphase CT, as a single imaging modality, provides a minimally invasive comprehensive pre operative evaluation that can be expected to have a major impact on patient selection and surgical planning. PMID- 11436578 TI - Minimally invasive techniques for the treatment of liver tumors. AB - Recently, several minimally invasive, image-guided therapies have been developed for the local treatment of hepatocellular carcinoma (HCC) and hepatic metastases in patients who are considered poor candidates for surgical resection. Radio frequency ablation, microwave ablation, and laser ablation all destroy tumor by creating a hyperthermic injury. Cryoablation and ethanol ablation achieve cellular death through freezing and direct toxicity, respectively. Chemoembolization is unique in that the entire liver can be treated over time with a combination of cytotoxic drugs and embolic agents, potentially reducing the rate of recurrence from radiologically occult tumor. Outcomes for minimally invasive therapy have approached the success rate of surgical resection in some series with considerably fewer complications. However, a paucity of randomized trials and variability in reporting limit assessment of the relative role of these techniques in clinical practice. With a few exceptions, the indications, contraindications, and complications associated with the different techniques are similar, and success with HCC has exceeded that for metastatic disease independent of technique. PMID- 11436579 TI - Right upper quadrant pain and fever in a patient with pulmonary sarcoidosis. PMID- 11436580 TI - [Goundou. Nasal bone yaws]. PMID- 11436581 TI - [Surinam, a key partner in public health regional cooperation in Guyana]. PMID- 11436582 TI - [Tropical destination. Stopver in Rodrigues]. PMID- 11436583 TI - [Human albinism: clinical, genetic, cellular, biochemical and molecular aspects]. PMID- 11436584 TI - [N-methyl glucamine antimoniate or Glucantime]. PMID- 11436585 TI - [Medical education in Madagascar from 1870 to present]. PMID- 11436586 TI - [Charles Merieux, an idealist impregnated with a profound sense of duty]. PMID- 11436587 TI - [Reinfection with Schistosoma haematobium and mansoni despite repeated praziquantel office treatment in Niger, Mali]. AB - The dynamics of reinfection by Schistosoma haematobium and Schistosoma mansoni after repeated treatment with praziquantel (40 mg/kg body weight, single dose) was studied in a cohort of schoolchildren living in an endemic area. A total of 214 urine and 220 stool samples were collected and examined at three different times, i.e., February 1989, July 1989 and February 1990. Mass chemotherapy was administered at the beginning of study (February 89). Treatment was repeated in children with positive tests at each subsequent sampling. Prevalence rates were 55.1 p. 100, 3.7 p. 100, and 35.0 p. 100 for Schistosoma haematobium and 62.7 p. 100, 46.3 p. 100 and 73.1 p. 100 for Schistosoma mansoni in February 1989, July 1989 and February 1990 respectively (p < 0.001). From July 1989 to February 1990, reinfection was observed in 84.5 p. 100 of children by Schistosoma haematobium versus 57.8 p. 100 by Schistosoma mansoni. The risk of reinfection by Schistosoma haematobium was higher in children between the ages of 7 and 10 years than in children between the ages of 11 and 15 years (p < 0.001), The incidence of intense Schistosoma haematobium egg excretion rose from 0 p. 100 in July 1989 to 6.0 p. 100 in February 1990. The incidence of intense Schistosoma mansoni excretion in February 1990 was 4.5 p. 100. The reinfection rate at 7 months was over 50 p. 100 for both parasite species despite repeated treatment. This finding demonstrates that additional measures such as proper sanitation and vector control are needed to control human schistosomiasis in irrigated rice paddies. PMID- 11436588 TI - [Cerebromeningeal listeriosis associated with a cytolytic hepatitis. First case report in Senegal]. AB - Hepatitis due to Listeria monocytogenes is uncommon in adults. This report describes the first case observed in Senegal. The patient was a 73-year old man presenting listeria-related hepatitis presumably secondary to low-grade meningeal encephalitis. Treatment using ampicillin was unsuccessful and the patient died four days after hospitalization. The authors note that the incidence of adult listeriosis has risen constantly for the past twenty years in relation with alcohol abuse, cirrhosis, diabetes, kidney insufficiency, cancer, AIDS, and organ transplantation. However no predisposing factors were observed in the present case. PMID- 11436589 TI - [Fish hooks: an unusual and sharp foreign body in the hypopharynx]. AB - Foreign bodies in the hypopharynx occur commonly. A myriad of objects have been observed. In this report we describe a case involving a fish hook in a 10-year old girl. The presence of this uncommon, sharp-pointed object was disclosed by plain radiography. The child was hospitalized three days after ingestion. Two attempts to perform endoscopic retrieval failed. Open cervicotomy was undertaken and allowed successful removal. The authors discuss management of this special type of foreign body and review several other unusual case reports in the literature. PMID- 11436590 TI - [Congenital immunodeficiency revealed by an acute anterior poliomyelitis. A case report]. AB - Acute anterior poliomyelitis is now uncommon in Algeria. Perusal of data from the health ministry shows no cases reported between 1990 and 1993. Since 1994 the incidence has increased slightly to 0.03 per 100,000 inhabitants in 1999. This report describes a case of acute anterior poliomyelitis due to type 2 poliovirus observed in 1996 in a six-year-old boy who had been immunized using oral polio vaccine in 1991. Hospital testing identified congenital immunodeficiency. The possibility that the vaccine caused the disease is discussed. PMID- 11436591 TI - [Current profile of hematogenous osteomyelitis in children at the Tokoin University Hospital Center in Lome, Togo. Report of 145 cases]. AB - Bacterial osteomyelitis is still a major health problem in Togo due to frequent association with sickle cell disease and delayed treatment. This report describes a series including 145 patients hospitalized for osteomyelitis in a university hospital center in Lome from 1995 to 1997. Osteomyelitis was acute in 33 cases, in association with sickle cell disease in 14 cases, and chronic in 112 cases including 103 with sequela of acute osteomyelitis. Data showed that the most common infectious agent was Staphylococcus aureus (64 cases) which was isolated or associated with another microorganism. The most valuable finding of this study was the high incidence of salmonella (n = 39) identified in association with sickle cell disease. If salmonella is detected, electrophoretic hemoglobin screening should be performed. PMID- 11436592 TI - [Spontaneous giant cephalohematoma in a 13-year-old child]. AB - Cephalohematoma is usually observed in the neonatal period often as a result of birthing injuries induced during difficult labor (vacuum extraction and forceps delivery). Less common causes include vascular abnormalities, aneurysm, arterial dissection, blood coagulation disturbances, and vascular wall frailty. This report describes a giant cephalhematoma with right ocular protrusion and anemia with no identifiable cause in a 13-year-old boy. Rapid recovery was achieved by surgical drainage and prompt dressing. PMID- 11436593 TI - [Mental confusion and multifocal lymphatic and pulmonary tuberculosis. A case report]. AB - This report describes a case of mental confusion associated with ongoing adenitis and pulmonary tuberculosis in a 20-year-old man with no history of psychiatric disorders. Diagnosis was based on clinical and laboratory findings. Tranquillizers improved mental status and antituberculosis treatment was administered before referring the patient to an internal medicine department. The authors emphasize the rarity of these cases in the Senegalese medical literature and discuss possible diagnostic pitfalls. PMID- 11436594 TI - [The new poor expatriates in the third world]. AB - This report describes four cases involving French expatriates who developed serious health problems while living destitute in Senegal. A 37-year-old man presented with embolism-like symptoms in relation with lung involvement due to spreading of untreated staphylococcal skin infection. A 64-year-old man was admitted for high output cardiac failure secondary to severe anemia caused by malnutrition. A 50-year-old man presented recurrent fever with loss of consciousness after successful treatment of neuromalaria and finally attributed to delirium tremens. A 25-year-old man was hospitalized with presumed meningoencephalitis that had been ongoing for two weeks. All four patients had been in Senegal for several months and had cut all ties to mainland France. Being jobless and homeless, they had been unable to obtain proper nutrition or medications. With minimal entry requirements and low-cost air travel, Senegal has been come a common sight-seeing destination and has developed a small but apparently growing population of destitute travelers. Alienation from the home community and resulting seclusion enhance clinical consequences and delay intervention by foreign service officials. PMID- 11436595 TI - [Did HIV 1 and the AIDS pandemic originate in central Africa during the past French colonization?]. PMID- 11436596 TI - [Value of the bilio-intestinal bypass in biliary disease in the Far East: retrospective study of 5 year experience at the Calmette Hospital of Phnom Penh (Cambodia)]. AB - Gallstone disease in the Far East exhibits a number of special features regarding epidemiology and etiology as well as indications for and response to surgical treatment. Surgical bypass techniques seldom used for treatment of gallstone disease in the West can be highly effective in lowering complication and recurrence rates in oriental patients. The purpose of this retrospective report is to describe a 5-year experience with gallstone disease at the Calmette Hospital in Phnom Penh (Kingdom of Cambodia). After reviewing the main pathological features encountered, we describe the indications for the two bypass techniques used. Choledocoduodenostomy was used for frail, elderly patients while choledochojejunostomy was the treatment of choice and achieved excellent short, middle, and long-term results in younger patients. PMID- 11436597 TI - [Value of intrathecal baclofen in the treatment of severe tetanus in the tropical milieu]. AB - Tetanus continues to be a cause of high mortality in developing countries, where resources for muscle relaxation and respiratory support are not readily available. Baclofen, a GABAB receptor agonist, directly restores physiologic inhibition of alpha motoneuron, that is blocked by tetanus toxin. Its use has been suggested at high dosage by intrathecal (i.t.) route, as part of the treatment of tetanus-induced contractures, and to limit the need for general anesthesia and for tracheal intubation in afflicted patients. This review reports personal experience and focuses on published data about i.t. baclofen for severe tetanus. Although statistical analysis are difficult to perform, i.t. baclofen appears to be effective in resolving muscle rigidity and in avoiding the need for deep sedation and for tracheal intubation, thus achieving lower mortality. Nevertheless, i.t. baclofen has a narrow therapeutic range, and a large interindividual pharmacodynamic variability. Thus, its use should be reserved for patients who would have been intubated without it. I.t. baclofen is a fairly simple and cost-effective modality, with significant advantages for management of severe tetanus, especially in tropical environment. PMID- 11436598 TI - [Inguinal hernia in Africa and laparoscopy: utopia or realism?]. AB - Inguinal hernia is a common indication for surgery in Africa. Most cases involve men and are treated in advanced stages often with complications. Until now the benchmark technique for surgical management has been the well-defined herniorrhaphy technique. Use of prosthetic implants has been rare because of high cost. Recently there has been a growing interest in video-assisted surgery throughout developing countries. However this enthusiasm should not obscure the fact that the technique is still in the developing stage and thus is more costly for the local economy. Indications for video-assisted surgery should be carefully selected in function of local conditions as well as problems specific to developing countries. PMID- 11436599 TI - [Ultrasonoic diagnosis of morbidity related to schistosomiasis due to Schistosoma mansoni and Schistosoma haematobium: epidemiological and individual value]. AB - By allowing study of large population samples, ultrasonography has revolutionized assessment of schistosomiasis-related morbidity. Previous clinical or parasitological parameters provided poor documentation of the public health impact of schistosomiasis. Thanks to a WHO-coordinated drive to standardize examination protocols and severity scores, comparison of data from different regions is now much easier. The latest "Niamey" methodology has eliminated the major shortcomings of earlier methods. Detection of periportal fibrosis is the cardinal diagnostic feature for Schistosoma mansoni. Ultrasonographic evidence has been validated by correlation with hepatic biopsy findings from hospitalized patients with severe disease. The specificity of ultrasonography is poor in low- or moderate-grade disease for which different methodologies give discordant results. Ultrasonography is highly sensitive for assessment of morbidity related to Schistosoma haematobium infection, which is associated with typical bladder lesions. Lesions involving the upper urinary tract are also well visualized but do not constitute a specific finding. The best applications for which ultrasonographic investigation of schistomsomiasis is now considered as mandatory are community-based studies and post-therapeutic follow-up of populations. In contrast ultrasonography is not well suited to individual diagnosis. In endemic areas, ultrasonography may be used for individual diagnosis if more effective methods are unavailable. However the poor specificity of some images is a major limitation for use in zones of low transmission. PMID- 11436600 TI - [Epidemiological and therapeutic aspects of plasmodial infection from Plasmodium vivax]. AB - A major challenge for successful treatment of Plasmodium vivax malaria is prevention of recurrence due to activation of dormant intrahepatic parasitic forms called hypnozoits. As a result of strain variability, recurrences are unpredictable and can occur months or even years after initial infection. Prevention requires elimination of both erythrocytic and hepatic parasite forms by combined use of chloroquine and primaquine. Primaquine is the only commercially available drug against hypnozoits. Several factors must be taken into account in planning treatment regimens. One factor is drug resistance of vivax strains, which as observed for Plasmodium falciparum strains is variable between geographical areas. Another factor is potential co-infection by Plasmodium vivax and falciparum, which is increasingly common. For optimal efficacy, treatment regimens must be adjusted with regard to dosage of primaquine and association with halofantrine, mefloquine or other new antimalarial agents. No regimen completely rules out the risk of recurrence. PMID- 11436601 TI - [Evolution over nine years (1989-1998) of HIV seroprevalence in pulmonary tuberculosis patients in Cameroon]. PMID- 11436602 TI - [Toxoplasma gondii in men and cats in Port-au-Prince (Haiti)]. PMID- 11436603 TI - [Role of non-validated treatments in the inter-critical management of asthma in adults in the African milieu]. PMID- 11436604 TI - [Value of a video questionnaire in the evaluation of the prevalence of asthma in Africa]. PMID- 11436605 TI - [Wild mammal trypanosomiasis in French Guyana: practical implications for the presence of Trypanosoma rangeli]. PMID- 11436606 TI - [Risk and economics of disease introduction to dairy farms]. AB - A more closed farming system will enhance the success of disease eradication programmes, because the introduction or re-introduction of infectious diseases is less likely. The objective of the study was to obtain input for the development of an on-farm decision support model to calculate the economic consequences of a more closed farming system. The input was based on bovine herpesvirus 1 (BHV1), since there were numerous data on this disease, but a more closed farming system will prevent introduction of other diseases as well (i.e. bovine virus diarrhoea virus (BVDV), L. hardjo, and S. dublin). Direct animal contacts, such as purchase of cattle, participation in cattle shows, and cattle that escape and mingle with other cattle, were found to be important risk factors for the introduction of BHV1. Furthermore, the use of protective farm clothing was found to be an important preventive factor. The effect of an IBR outbreak at an IBR-free farm on milk production caused limited losses of on average 0.9 kg per cow per day during 9 weeks, but the variability was high (95% CI 0-2 kg). Nine percent of Dutch IBR free dairy farms that were also at risk for BVDV, L. hardjo or S. dublin had one introduction per year of one of these four diseases. All these results were incorporated in the economic model. Management measures to reduce the probability of introduction of BHV1, the costs of these measures, and the risk reduction after these measures were obtained from other sources. The calculations showed that the implementation of a more closed system will be profitable for most farms. The profitability will increase when a farm is at risk for more diseases, but will decrease when farms are limited in their facilities to rear replacement heifers or when a large proportion of pasture adjoins pasture of other cattle farms. PMID- 11436607 TI - [Ataxia and pruritus in a pony due to a cervical vertebral fracture]. AB - A mini-Shetland stallion was referred with bilateral pruritus localized to the anterior neck. More detailed clinical examination also revealed ataxia, and diagnostic imaging, including pre- and post-contrast computed tomography, revealed an old fracture of C2. Spinal cord compression was diagnosed. The probable causal relation between the fracture, the ataxia, and the localized pruritus seems comparable to the 'contusio cervicalis posterior' in humans, in which spinal cord trauma may cause pruritus in the associated dermatome. PMID- 11436608 TI - [Report from the Wijffels committee about reorganizing the sector. Future of farming: a challenge for the farmer]. PMID- 11436609 TI - [Interview with William Schaftenaar, veterinarian at Blijdorp zoo. 'I think you need to ask yourself what role you want to play in society']. PMID- 11436610 TI - [Meeting of the Panel on Food and Nutrition of Small Animals. Selling of animal feed by veterinarians is logical]. PMID- 11436611 TI - [Legal liability of the owner of a horse for injury to a veterinarian]. PMID- 11436612 TI - [Committee for Recommending Veterinary Products handles two complaints. Public advertisement of injectable antibiotic and brand misuse]. PMID- 11436613 TI - [Suspicion of foot and mouth disease in Dutch wildlife]. PMID- 11436614 TI - [Closing of cattle markets]. PMID- 11436615 TI - [Committee of 'the Crying Pig' erect memorial]. PMID- 11436616 TI - [Position of the Group for Homeopathic-working Veterinarians]. PMID- 11436617 TI - [Enormous excitement justified]. PMID- 11436618 TI - [Via kindergarten to reconsideration?]. PMID- 11436619 TI - [The relative risk of the Spring Days, a reaction to Klaare, et al]. PMID- 11436620 TI - [Money from our wallet]. PMID- 11436621 TI - Adult protection legislation in Canada. Models, issues, and problems. PMID- 11436622 TI - Health in the 'grey' millennium. Romanticism versus complexity? PMID- 11436623 TI - Finding the elder voice in social legislation. PMID- 11436624 TI - Aging, homelessness, and the law. PMID- 11436625 TI - The abuse and neglect of the elderly. PMID- 11436626 TI - Consumer choice in long-term care. What the United States can teach and learn from others about decisionally incapacitated consumers. PMID- 11436627 TI - The emergence of older adults' personal relationships in Canadian law. PMID- 11436628 TI - Being "reasonable." Defining and implementing a right to community-based care for older adults with mental disabilities under the Americans with Disabilities Act. PMID- 11436629 TI - The strange interplay of testamentary capacity and the doctrine of undue influence. Are we protecting older testators or overriding individual preferences? PMID- 11436630 TI - Loss of competency in Alzheimer's disease: conceptual and psychometric approaches. PMID- 11436631 TI - Elders and criminal justice. International issues for the 21st century. PMID- 11436632 TI - Coping with crime. An examination of elderly and nonelderly adaptations. PMID- 11436633 TI - Elders in prison. Health and well-being of older inmates. PMID- 11436634 TI - Globalisation and guardianship harmonisation or (postmodern) diversity? PMID- 11436635 TI - [Sequencing and comparative analysis of the complete glycoprotein gene of three Crimean-Congo hemorrhagic fever virus Chinese isolates]. AB - OBJECTIVE: To determine the complete sequences of glycoprotein (M) gene of Crimean-Congo hemorrhagic fever virus (CCHFV) Chinese isolates (Xinjiang hemorrhagic fever virus, XHFV) BA66019, BA8402 BA88166 and to compare and analyze the relationships among the viruses. METHODS: Based on the conserved reverse complementarity of the CCHFV genomic termini, a primer PCM-Tag was designed and used together with random hexmers to initiate reverse transcription of viral RNA to synthesize cDNA. The latter was amplified with single primer PCM-Tag using the proof-reading DNA polymerase to produce the complete M gene. The PCR products were gel-purified and the whole M segment was sequenced and the comparison and analysis were performed aided by computer for the phylogenesis and coding strategy. RESULTS: Comparison of the whole M gene sequences of XHFV reference strain BA66019 with international prototype CCHFV IbAr10200 showed an excess of 5 base pairs in length, resulting in 5,365 bp; while BA8402 and BA8816 are 5,365 bp long, 4 more base pairs than IbAr10200. The first start codons of long ORFs were located at the 78th base pair in M gene of BA66019, BA8402 and BA88166, which were 15 base pairs prior to that of IbAr10200. The precursors encoded by these M genes were totaled 1,689 amino acids (AA), 6 AAs more than that of IbAr10200. The similaries between CCHFV M genes in nucleotide level were: 80.9% (IbAr10200 BA66019), 80.2% (IbAr10200-BA8402), 80.2% (IbAr10200-BA88166), 83.7% (BA8402 BA66019), 83.6% (BA88166-BA66019), and 99.0% (BA8402-BA88166), respectively. The corresponding similarities in amino acid level were 85.1%, 86.3%, 86.6%, 87.8%, 88.0%, and 98.8%, respectively. The similarities of M genes of CCHFVs and Dugbe virus in both nucleotide and amino acid levels were around 55% and 37%, which were much lower than those among CCHFVs. CONCLUSION: The M genes of XHFVs and IbAr10200 formed respective independent phylogenetic branches genetically and the human-origin isolate BA88166 might be a variant of tick-borne BA8402, suggesting that there was only one kind virus prevailing in Xinjiang epidemic areas in 1980s. PMID- 11436636 TI - [Complete genome sequence analysis of the Hantavirus Z10 strain]. AB - OBJECTIVE: Study on the complete genome sequence of Hantavirus Z10 strain which has been applied for inactivated vaccine production in China, to assess its molecular characteristics and the diversity with other hantaviruses. METHODS: The total RNA were prepared from Z10 virus infected cells and the RT-PCR products was cloned into T vector, sequenced and analyzed by using DNASTAR software. RESULTS: The Z10 complete genome, L segment is 6,553, M segment is 3,615, S segment is 1,701 nucleotides in length, with a single open reading frame encoding 2,151, 1,135, 429 amino acids respectively. Sequence homology comparison showed that the 3 segment nucleotide of Z10 strain were close to HTN type virus, but only 83.6 87.4% homology with other HTN viruses at the nucleotide level. The phylogenetic analysis was made on their nucleotide and amino acid sequences. CONCLUSION: The results firstly demonstrates that Z10 strain is a new subtype of the Hantaan(HTN) type. PMID- 11436637 TI - [Cloning and expression of human TNFR (P55)-IgG Fc fusion protein in eukaryotic cells]. AB - OBJECTIVE: To express soluble biologically active TNF receptor protein in eukaryotic cells, which can inhibit cytotoxic activity of TNF. METHODS: PCR amplified the extra-cellular region of TNF receptor P55 and IgG Fc gene. Then the two were linked through an oligomer encoding a thrombin-sensitive peptide linker and cloned into eukaryotic expression vector pcDNA3.1 (+). The eukaryotic expression plasmid pcDNA3. 1/TI was then transfected to the mammalian cell line COS7 and BHK. Using the G418 system, BHK cell clones were selected and can continuously secrete biological protein in large amount. RESULTS: The expressed protein was fused with IgG Fc and secreted into the cell culture supernatant. It has good antigenicity and binding ability to TNF. It can also inhibit the cytotoxic activity of TNF on L929 cell. CONCLUSION: The TNFR-IgG Fc fusion protein expressed in eukaryotic cells has biological activities of human TNF receptor P55. PMID- 11436638 TI - [Two virus strains of isolated in China are recombinant alphaviruses]. AB - OBJECTIVE: To determine the classification, phylogenetic and genetic type of XJ 90260 and XJ-91006 viruses isolated in China. METHODS: Two pairs of specific primers were designed to amplify the NSP4 and E1-3' UTR genes of XJ-90260 and XJ 91006 viruses and the nucleotide sequences between them and with that of other Alphavirus were compared. RESULTS: The nucleotide sequences identity between XJ 90260 and XJ-91006 viruses was 100% and has showed the highest with WEE, comparing with the other 26 Alphaviruses in 3'-UTR. NSP4 gene of XJ-90260 and XJ 91006 viruses high homologous with EEE virus and E-1 gene high homologous wih SIN virus. XJ-90260 and XJ-91006 viruses belong to the B group of WEE and are closest to the Russian strain (Y62-33) by phylogenetic analysis. CONCLUSION: XJ-90260 and XJ-91006 viruses isolated in China are recombinant Alphaviruses, they belong to the same genetic type as WEE does. PMID- 11436639 TI - [Study on interactions between TPO molecules tested with yeast two-hybrid system]. AB - OBJECTIVE: To explore the interactions between TPO molecules and their locations. METHODS: Full-length TPO, TPO(N) and TPO(C) were inserted respectively into the plasmids of the yeast two-hybrid system. Six different recombinant plasmids were generated. Every 2 recombinant plasmids were transformed into the yeast cell SFY526. The possible interactions between TPOs were then tested by the bioactivity of beta-galactosidase. RESULTS: There are interactions between full length TPO themselves TPO(N) themselves and TPO with TPO(N), but not between TPO(C)themselves, TPO with TPO(C) and TPO(N) with TPO(C). CONCLUSION: Naturally, the molecular interaction domain of TPO may exist in the N terminus but not in the C terminus. PMID- 11436640 TI - [The effect of EB virus bhrfl gene expressing on the topoisomerase I expression in nasopharyngeal carcinoma cell line]. AB - OBJECTIVE: To study the effect of bhrfl expressing on the topoisomerase I expression in nasopharyngeal carcinoma cell line CNE2. METHODS: bhrfl recombinant expressing vector was constructed and used to transfect the nasopharyngeal carcinoma cell line CNE2. The CNE2 cells were then tested for changes of the topoisomerase I expression and anti-apoptotic ability under the treatment of camptothecin. RESULTS: bhrfl expressing could enhance the topoisomerase I expression in CNE2 cells, that in end reduced cell sensitivity to camptothecin increased cell survival span and decreased apoptotic rate(P < 0.01). CONCLUSION: By increasing the expression of topoisomerase I, bhrfl expressing can enhance the resistance of CNE2 cells to apoptosis induced by camptothecin. PMID- 11436641 TI - [Preliminary analysis of genetic characteristics of the nonstructural protein NSP4 of rotavirus strains with different serotypes in China]. AB - OBJECTIVE: To study the genotypes and genetic variations of nonstructural protein NSP4 of rotavirus strains with different serotypes in China. METHODS: NSP4 cDNA of the four serotypes G1, G2, G3 and G9 were cloned and sequenced by SILVER SEQUENCE DNA Sequencing System. RESULTS: The identity of the nucleotide and amino acid were 78.8%-93.5%, 83.0%-95.5%, respectively. NSP4 from strains G1, G3, G9 all fell into Wa allele, while that of G9 fell into KUN allele. Some variations of the amino acids were observed localizing at the position of 135-145 in the VP4 binding domain. CONCLUSION: The results indicated that the two alleles Wa and KUN are the major prevalent NSP4 genotypes in China. PMID- 11436642 TI - [Experimental study of preventive effect of xin-kang oral liquid on acute viral myocarditis in mice]. AB - OBJECTIVE: To study the preventive effect of Xin-Kang oral liquid on acute Coxsackie B3 viral myocarditis in mice. METHODS: The mice were infected Coxsackie virus B3 (CVB3m) through abdominal cavity after they were fed with the Xin-Kang oral liquid for 2d, and the uses of the drug continued to the 20th day. Histopathological changes of the myocardial cells were observed under microscope and measured by morphometric quantification. RESULTS: The detection rate of pathological change of grade III of heart surface in Xin-Kang oral liquid groups were lower than in virus control group, the difference between Xin-Kang medium dose group and virus control group being specially significant (P < 0.01), and the detection rate of severe myocardial damage was much lower. The average area of myocardial damage and area of myocardial damage/area of the whole heart in Xin Kang oral liquid groups were markedly lower than those in virus control at the 5th day and the 20th day. CONCLUSION: These results demonstrate that the preventive effect of Xin-Kang oral liquid on acute Coxsackie B3 viral myocarditis is similar to that of interferon. PMID- 11436643 TI - [An inquiry into the relationship among pathology, clinic and laboratory indexes of the severe viral hepatitis]. AB - OBJECTIVE: This study was to analyze the relationship among pathology, clinic and laboratory indexes in 62 cases of the acute severe hepatitis (ASH) and 72 cases of the subacute severe hepatitis(SSH) to offer a correct diagnosis and recognition of the disease. METHODS: A total of 134 severe hepatitis cases with confirmed pathology treated in a course of 24 years were analyzed by programs of EXCEL 2000. RESULTS: When compared ASH with SSH, the onset of the age were 40 +/- 13 years old and 44 +/- 15 years old, P < 0.05, the rates of hepatitis coma were 100% and 66.7% respectively. The liver weight of all the cases decreased. The tests of TBiL, PTA, Cho, NH3 and WBC between ASH and SSH had significant differences. CONCLUSION: Hepatic encephalopathy is a necessary criterion for ASH. The TBiL, PTA and Cho changes are closely related with severe viral hepatitis, they not only are the essential laboratory indexes but also the indispensable factors for disease development and prognosis of severe viral hepatitis. PMID- 11436644 TI - [Clinical efficacy of lamivudine in the treatment of chronic hepatitis B]. AB - OBJECTIVE: To study the clinical efficacy and safety of lamivudine in the treatment of chronic hepatitis B. METHODS: 72 patients with hepatitis B were randomly assigned into lamivudine (n = 54) and placebo groups (n = 18) for 12 weeks. Then, all the patients received lamivudine 100 mg daily until week 104. Clinical symptoms, liver function tests, serum HBV DNA and YMDD mutation were evaluated. RESULTS: HBV DNA response rate of lamivudine group was higher than that of placebo group after 12 weeks treatment (61% vs 6%, P < 0.01). ALT sustained normalization rate of lamivudine group was higher than that of placebo group (65% vs 11%, P < 0.05). At week 52 and week 104, HBV DNA response rate was 78% and 36% respectively, whereas the normalization rate of ALT was 39% and 33% respectively. Proportion of HBeAg/anti-HBe seroconversion was both 6.1% at week 52 and 104. Overall YMDD mutation rate was 13.7% at week 52 and 39.7% at week 104. The incidence of adverse effects was similar for lamivudine and placebo group at week 12. There was no severe drug-related adverse events during 104 weeks treatment. CONCLUSION: Lamivudine 100 mg daily could suppress HBV replication rapidly and be well tolerated. However, in some cases, YMDD mutation may lead to HBV DNA breakthrough. PMID- 11436645 TI - [Inhibition of HBV gene expression by antisense oligonucleotides using galactosylated poly (L-lysine) as a hepatotropic carrier]. AB - OBJECTIVE: To study the specific inhibition of HBV gene expression by antisense oligonucleotide (ASON) targeted by galactosylated poly (L-lysine) (Gal-PLL). METHODS: According to the results of direct sequencing of PCR amplified products, a 16 mer phosphorthioate analogue of the antisense oligonucleotide (PS-ASON) directed against the HBV U5-like region was synthesized and then linked with one liver-targeting ligand, the Gal-PLL. Using the 2.2.15 cells compared the effect of them on the expression of HBV gene. RESULTS: We identified that HBV DNA in the 2.2.15 cells was from HBV with surface antigen subtype ayw2 by sequencing. The fluorescent histochemistry test indicted that Gal-PLL had a selective affinity to the rat liver tissues. A 2:1 molar ratio of the Gal-PLL to DNA optimized the complex formation. In the same experimental conditions, the inhibitory effects of HBsAg and HBeAg by PS-ASON were 70% and 58%, respectively at a concentration of 10 mumol/L, while by ligand-PS-ASON were 96% and 82%, respectively, and the amount of HBV DNA in culture supernatant and cells was depressed significantly. An unrelated sequence oligonucleotide showed no effectiveness. All the oligonucleotide had no cytotoxicity. CONCLUSION: Antisense oligonucleotides complex with the liver-targeting ligand can be targeted to cells via asialoglycoprotein receptors resulting in specific inhibition of HBV gene expression and replication. PMID- 11436646 TI - [Effects of attenuated measles virus on hepatitis B virus in vivo and in vitro]. AB - OBJECTIVE: To observe the effects of attenuated measles virus on hepatitis B virus replication inside and outside the body. METHODS: Patients with chronic hepatitis B virus replication were chosen, and attenuated measles viruses were inoculated in them and observed twice every week for a total of 48 weeks. The changes of CD4+, CD8+, IFN-gamma, IL-2, HBeAg and HBV DAN were observed. Also the changes of the HBVM were observed when adding the measles virus in suspension of the 2.2.15 cells. RESULTS: The average ratio of CD4+ and CD8+ before the treatment was 1.05 +/- 0.19 and it rose to 1.47 +/- 0.18 after follow up for 48 weeks. Then, compared with the ratio before the treatment, the difference was significant(t = 5.36, P < 0.01). After inoculating the measles virus for two weeks, IFN-gamma and IL-2 increased. Although the total average ratio had not great difference, IL-2 level of 26.3% of the cases elevated 5 times higher in the second week, as compared with the control group, there was a great difference (chi 2 = 12.07, P < 0.005). The results of the examination of the HBeAg before treatment was 317, 35 +/- 151.75, but became 139.17 +/- 166.66 after observing for 48 weeks(t = 2.16, P < 0.05). The positive ratio of HBV DNA before the treatment was 100% and become 63.16% after treatment. Thus the ratio difference is significant(chi 2 = 10.08, P < 0.005). However, no such ratio difference was found in the control group. When adding the attenuated measles virus in suspension of 2.2.15 cell, the HBsAg and HBeAg decreased. CONCLUSION: When the measles virus and hepatitis B virus infect the cases in an overlapping way, the latter may be interfered by change of the cell immunity status of the body of the former, and measles virus may have direct interference effects on hepatitis B virus. PMID- 11436647 TI - [Detection of anti-HAV IgM and anti-HEV IgM in patients with sporadic acute hepatitis in Beijing between 1995-2000]. AB - OBJECTIVE: To investigate the age ranges, liver function lesion and outcomes of sporadic acute hepatitis A and E in Beijing. METHODS: Enzyme immunoassay(EIA) was used for detection of anti-HAV and anti-HEV immunoglobulin M(IgM). Samples were collected from the patients with sporadic acute viral hepatitis in Beijing between 1995-2000. RESULTS: The total rate of positive for anti-HAV and anti-HEV was 55.2% (112) in 203 patients with acute hepatitis, 22.2% (45) and 33% (67) were positive separately for anti-HAV and anti-HEV in 112 patients. The duration of anti-HEV IgM was 45-60 days and that of anti-HAV IgM was at least 2-3 months. The patients with acute hepatitis A and hepatitis E all experienced jaundice, a rising of liver enzyme, but not developed to chronic hepatitis or to death. CONCLUSION: Acute hepatitis A as well as acute hepatitis E takes an important place in sporadic enterically transmitted hepatitis in Beijing. PMID- 11436648 TI - [Establishment of ELISA with TT virus(TTV) and its application to patients with hepatitis and healthy people]. AB - OBJECTIVE: To establish the EIA method for detection of antibody to TT virus (anti-TTV) and to investigate the distribution of anti-TTV in patients with hepatitis and in healthy people. METHODS: The ELISA method using the recombinant expressed product of truncated gene of TTV ORF1 as antigen was developed and was applied to detect anti-TTV in patients with hepatitis and in healthy control. The TTV DNA were detected by nested-PCR. RESULTS: The positive rates of anti-TTV antibodies in different populations were as follows: 1.3% in healthy people, 10.5% in patients with hepatitis A, 12.5% in patients with hepatitis B, 8.3% in patients with hepatitis C, 7.7% in patients with hepatitis D, 12.0% in patients with hepatitis E, 6.5% in patients with hepatitis G, 32.3% in patients with hepatitis non-A to G. The positive rate of TTV DNA was significantly related to that of anti-TTV (P < 0.05). CONCLUSION: The anti-TTV antibodies were detected positive in patients with hepatitis and healthy people, however, its positive rate in patients with hepatitis non-A to G was significantly higher than that in other groups. The fact that anti-TTV antibodies coexist with TTV DNA suggests that the antibody to TTV ORF1 antigen may be an infectious marker of TTV similar to that of anti-HCV. PMID- 11436649 TI - [Study on p53 gene mutation in hepatocellular carcinoma patients in China]. AB - OBJECTIVE: To study p53 gene mutation in human hepatocellular carcinoma (HCC) samples and the relationship between p53 gene mutation and hepatitis B virus (HBV) infection. METHODS: DNA samples were prepared from 50 specimens of HCC patients that had been infected with HBV. Exons 5-9 were amplified with polymerase chain reaction (PCR), and then detected by single strand conformation polymorphism (SSCP). RESULTS: Over 26% (13/50) of p53 DNA samples were found to be mutated, mainly in exons 5-8 relating to 3,3,4,3 cases respectively and along with 4 suspicious samples. CONCLUSION: p53 gene mutation might be one of the causes of HCC, and HBV infection may be associated with such mutation in China. PMID- 11436650 TI - [Clinical study on the relationships between HBV C gene promotor gene mutation and immunological marker, HBV DNA concentration]. AB - OBJECTIVE: To investigate the relationships between HBV infected patients immunological marker, HBV DNA concentration, and clinical symptoms. METHODS: PCR microplate nucleotide acid hybridization-ELISA and FQ-PCR methods were applied in the analysis of HBV BCP gene mutation and HBV DNA concentration in 246 HBV infected patients. RESULTS: In patients with HBsAg+/HBeAg+/anti-HBc+, HBsAg+/anti HBeAb+/anti-HBcAb+, and HBsAg+/anti-HBc+, the HBV DNA positive rate were 97.9% (48/49), 22.9% (25/109), 31.1% (14/45) respectively, with the overall positive rate 42.85% (98/203). And the HBV BCP gene mutation rate were 59.1% (29/49), 11% (12/109), 13.3% (6/45) respectively, with the overall mutation rate 23.15% (47/203). In HBV BCP gene mutation group HBV DNA concentration of each specimen was more than 10(6) cps/ml, which are significantly higher than other groups(P < 0.001). CONCLUSION: Re-evaluation of HBV infected patients with HBsAg+/anti HBe+/anti-HBc+ and HBV BCP gene mutation is necessary. The immunological turnover of HBeAg indicates the immune clearance and rest of HBV only in part of HBV infected patients. The immunological marker should not be the sole factor upon which the HBV replication status, the diagnosis and prognosis of patients are determined. PMID- 11436651 TI - [Application of dhfr gene negative Chinese hamster ovary cell line to express hepatitis B virus surface antigen]. AB - OBJECTIVE: To set up an efficient expressing system for recombinant hepatitis B virus surface antigen (HBsAg) in dhfr gene negative CHO cell line. METHODS: HBsAg gene expressing plasmid pCI-dhfr-S was constructed by integrating HBsAg gene into plasmid pCI which carries dhfr gene. The HBsAg expressing cell line was set up by transfection of plasmid pCI-dhfr-S into dhfr gene negative CHO cell line in the way of lipofectin. RESULTS: Under the selective pressure of MTX, 18 of 28 clonized cell lines expressed HBsAg, 4 of them reached a high titer of 1:32 and protein content 1-3 micrograms/ml. CONCLUSION: In this study, the high level expression of HBsAg demonstrated that the dhfr negative mammalian cell line when recombined with plasmid harboring the corresponding deleted gene can efficiently express the foreign gene. The further steps toward building optimum conditions of the expressing system and the increase of expressed product are under study. PMID- 11436652 TI - [The effect of IFN-alpha 2b-thymus peptides combined therapy in chronic hepatitis B]. AB - OBJECTIVE: To determine the effects of IFN-alpha 2b associated with thymus peptides in treatment of chronic hepatitis B. METHODS: A total of 52 individuals with chronic hepatitis B received IFN-alpha 2b(5 x 10(6) IU/ml qod) for 6 months and thymus peptides(20 mg qod) for 3 months. 50 patients received SMMC or other medicine such as WuWeizi, CuiPenCao served as control. There were no differences between two groups in sex, age, duration and seriousness of the disease. RESULTS: The primary outcome measure for efficacy was the percentage of HBV DNA/HBeAg negativity after 6 months therapy. Rate of HBV DNA negativity was 75% and that of HBeAg was 73.1%, the inhibitory activity of IFN-alpha 2b against HBV was significantly higher than that of the control. Except transient fever, leukopenia and thrombocytopenia appearing at the beginning of the therapy, no serious adverse events were observed. CONCLUSION: Potent inhibitory activity against HBV of IFN-alpha 2b is suggested and much works need to be done to improve the long term effect of the therapy. PMID- 11436653 TI - [Clinical implications of HCV quasispecies heterogeneity in patients with hepatitis C]. AB - OBJECTIVE: To determine the heterogeneity of viral quasispecies and its clinical significance in patients with hepatitis C. METHODS: Quasispecies in the sera from 76 patients infected with hepatitis C virus were detected using single stranded conformational polymorphism (SSCP) analysis of the HCV E2 hypervariable region 1 (HVR1). RESULTS: HVR1 was amplified in 72 (94.7%) of the 76 patients. The average number of SSCP bands was 5.8, with a range from 2 to 11. The numbers of quasispecies in acute hepatitis, chronic hepatitis and liver cirrhosis and/or primary hepatocellular carcinoma were 3.1 +/- 1.2, 6.0 +/- 2.3 and 8.4 +/- 4.1, respectively. There was a statistically significant difference among them (P < 0.01). Patients with infection acquired by blood transfusion and i.v. drug use had greater number of quasispecies than those acquired by other transmission pathway (sporadic) (P < 0.05). Patients with genotype 1 a and 1 b infection had increased quasispecies compared with those infected with HCV type 2 and 3(P < 0.05). Increased quasispecies heterogeneity was significantly correlated with serum HCV RNA levels (P < 0.01). CONCLUSIONS: SSCP is a simple, rapid and reliable method for the analysis of HCV viral quasispecies heterogeneity. Quasispecies heterogeneity plays an important role in HCV persistent infection and in the progress of hepatitis C. The duration of HCV infection, HCV genotype and HCV viremia are important determinants for the evolution of HCV quasispecies heterogeneity. PMID- 11436654 TI - [Detection of TORCH genom in the cardiac tissue of congenital heart disease]. AB - OBJECTIVE: To explore relationships of TORCH (cytomegalovirus/CMV, toxoplasma/TOX, rubella virus/RV, herpes simplex virus-2 HSV2) infection with congenital heart disease (CHD). METHODS: We conducted comparison to detection TORCH pathogenic gene in 42 cases of CHD and 38 controls by polymerase chain reaction (PCR). RESULTS: 1. The positive rate of CMV, HSV2 and TOX in 66 cases of CHD were 26.2%, 4.7% and 16.7%, respectively, while those of 38 cases in control groups were 21.1%, 2.6% and 2.6%, respectively. The infection condition of TOX was significantly different (P = 0.0378), while those of CMV and HSV were not statistically different (P > 0.05) between CHD and control groups. 2. 7 of 30(23.3%) were positive for RV-RNA, compared with control group which were all negative, it was significantly higher (P = 0.0328). CONCLUSION: This is the first report of the presence of TORCH genes in CHD cardiac tissue by PCR. It might lay the foundation for studying the relationship of TORCH infection with CHD in molecular level. PMID- 11436655 TI - [Quantitative analysis of cell fusion caused by glycoprotein of paramyxoviruses with reporter gene method]. AB - OBJECTIVE: To establish a kind of quantitative assay method for analyzing the reporter gene assay for fusion was established. METHODS: Two populations of BHK21 cells were infected with vTF-7 recombinant and wild-type vaccinia viruses respectively and transfected with paramyxovirus F and HN cDNA and plasmid pG1NT7 beta-gal respectively. After 16 h, the two cell populations were removed from the wells by trypsinization. Equal numbers of the two populations were mixed in wells of a 96-well flat-bottom plate after washing and pelleting. After 15 hours' incubation at 37 degrees C, the cells were lysed with Nonidet P-40 and A values were read at 570 nm with an ELISA reader supported by SOFTMAX software. RESULTS: Reporter gene method had a close relationship with cell count method, r = 0.9890(P < 0.01). Positive and negative coincidence rates were 100%. The best main reaction conditions are as follows: substrate concentration, 16 mmol/L; color reaction time, 20-25 min; amount of DNA for transfection, 1 microgram; cell numbers for each population, 1 x 10(5). CONCLUSION: Reporter gene method is a kind of very sensitive, specific and repeatable quantitative analysis for cell fusion and can be used in the studies on membrane fusion caused by any viruses. PMID- 11436656 TI - [Generation of human recombinant antibody Fab fragment to influenza virus from combined antibody library]. AB - OBJECTIVE: To acquire the recombinant human monoclonal antibodies to influenza virus which can be used in mucous membrane treatment to prevent the infection of virus in animal model in the future. METHODS: Construct and pan human Fab antibody library to influenza virus Sydney strain (H3N2) from vaccinated donors by using phage display technology and secreted expression in E. Coli system. The positive Fab clones with neutralization activity to influenza virus were selected and characterized through HI test, IFA and VN test. RESULTS: Separate two Fab clones, IV-2 and IV-6, appear positive responses in ELISA and IFA. These two clones can inhibit hemagglutination in HI test and can decrease viral titer by 30 and 20 times in viral neutralization assay, respectively. CONCLUSION: We have got two clones of Fab fragments with neutralization activity to influenza virus Sydney strain (H3N2). PMID- 11436657 TI - [The preparation of human single chain Fv antibody specifically against hepatitis C virus NS3 antigen and its application in immunohistochemistry]. AB - OBJECTIVE: Immunohistochemistry identification of human single chain Fv antibody (ScFv) against hepatitis C viral NS3 antigen. METHODS: The recombinant phages were panned by NS3 antigen which was coated on a microtiter plate, after five rounds of biopanning, 66 phage clones were identified specific to NS3 antigen. The affinity and specificity of ScFv were evaluated by ELISA, dot blot hybridization and immunohistochemistry. RESULTS: The data of ScFv-NS3 DNA digestion and DNA sequencing showed that the ScFv gene is composed of 747 bp. ELISA, dot blot hybridization and immunohistochemistry demonstrated that the human single chain Fv antibody against hepatitis C NS3 antigen has a specific combination character with hepatitis C virus NS3 antigen of different sources and paraffin-embedded patients tissues, it didn't react with liver tissue and HBsAg. CONCLUSION: The application of HCV NS3 specific ScFv in immunohistochemistry was successfully achieved. PMID- 11436658 TI - Histopathology of feral fish from a PCB-contaminated freshwater lake. AB - The purpose of this study was to evaluate the potential toxic effects of chronic sublethal polychlorinated biphenyl (PCB) exposure on feral fish, using histopathology as an endpoint. Histopathological study of bream (Abramis brama) and asp (Aspius aspius) living in a PCB-polluted freshwater lake revealed abnormal cellular changes in the renal corpuscle of both species. Dilation of glomerular capillaries (DGC), mesangial edema (ME), an adhesion between visceral and parietal layers of Bowman's capsule (ABC), and filling of Bowman's space (FBS), were highly prevalent features in lake fish. The prevalence of each of these lesions was significantly lower, or totally absent in fish caught from reference locations. Cellular alterations in liver, gill, gonads, spleen, and intestine were all linked to seasonal changes. The results suggest that some of the observed histopathological changes in renal glomeruli, particularly DGC and ME, could possibly indicate a prolonged chemical stress caused by PCBs and related compounds. It is also possible that chronic PCB exposure may have suppressed and weakened the immuno systems of exposed fish making them more vulnerable to secondary parasitic infections. PMID- 11436659 TI - Quantification of anthropogenic threats to lakes in a lowland county of central Sweden. AB - An evaluation of the negative effects caused by anthropogenic influence on lake ecosystems was performed, using data from 143 catchments in Uppsala County, Sweden. The study included i) technical encroachments; i.e. construction of dams, dikes, etc. as well as effects of drainage of land; ii) pollution, i.e. eutrophication, acidification, and contamination by toxic substances; iii) introduction of nonnative species; and iv) exploitation of species populations. Severe damage was caused mostly by drainage of land followed by pollution by toxic substances and, to a smaller extent, introduction of nonnative species and eutrophication. Most lakes were subject to several types of disturbances, interacting in a complex pattern, which made it difficult to link the visible effects to the true causes of the disturbance. Future lake management should include analyses of all disturbances to the lake catchments, taking into account the historical perspective, in order to balance the threats/damages, in an analysis of the possibilities for maintaining biodiversity and sustainability in the ecosystems. PMID- 11436660 TI - Sediment surface geochemistry of three Baltic Sea Deep Basins. AB - Three deep basins were chosen for this study and the most recent changes that are reflected in the uppermost 15 to 35 cm are reported. The Gotland Basin, the largest continuous sedimentation basin in the southern Baltic Sea, was chosen for the present study. Simultaneously, a quite large basin, the North Central Baltic Basin was investigated and chemical data are reported for the first time. In the southern Baltic Sea the Bornholm Basin was investigated. All 3 basins are different as regards the physicochemical conditions in the area. The investigated elements were divided into different groups according to their origin and geochemical behavior: elements of natural terrestrial origin; nutrients; elements that are confined to early diagenetic minerals; and in the near-surface zone the anthropogenic elements including several harmful metals. Although there seems to be a decreasing trend in the surface zone the metal concentrations are still clearly elevated. PMID- 11436661 TI - Sustainability of coastal resource use in San Quintin, Mexico. AB - San Quintin, Mexico, provides a useful site for integrated analyses of material fluxes and socioeconomic constraints in a geographically isolated system. Natural resource utilization on the land is dominated by groundwater exploitation for cultivation of horticulture crops (primarily tomatoes). Irrigation exceeds water recharge minus export by a factor of 6. Resource utilization in the bay is dominated by oyster culture; food for the oysters is provided by tidal exchange of bay and ocean water. Consideration of oyster respiration and system respiration suggests that the present level of aquaculture is about 40% of the sustainable level. A "physical unsustainability index" (PhUI) was developed to measure the proportional departure of utilization of the most limiting resource for sustainability: 6 on land; 0.4 in the bay. Based on PhUI and measures of economic development, we conclude that aquaculture is more viable than agriculture. PMID- 11436662 TI - Scientists speculate on what the Swedish scientist Svante Arrhenius would have worked on today. PMID- 11436663 TI - Social cost-benefit analysis of a watershed development project in Karnataka, India. AB - Watershed Development Programmes (WDPs) have been initiated in India to improve and sustain productivity and the production potential of the dry and semiarid regions of the country at higher levels, through adoption of appropriate production and conservation techniques. The aim is also to meet the needs of rural communities for food, fuel, fodder, and timber and, thereby, reduce pressure on natural forests. In view of their potential for growth, for improving income, and the natural resource base of the disadvantaged regions of the country, WDPs are being accorded importance in the development plans for India and by donor agencies. This paper presents a social cost-benefit appraisal of a watershed development project in Karnataka, India. Using alternate viability measures, i.e. Net Present Value (NPV), Benefit-Cost Ratio (BCR) and Internal Rate of Return (IRR), and rigorous tests and sensitivity analyses, the results show that if expected full benefits are realized, the benefits derived from the watershed project are quite high, with the IRRs ranging from 19 to 96%. If, however, expected full benefits were to fall short by 25%, and net costs of (including the opportunity cost of grazing benefits foregone by the villagers on account of establishing community woodlots on degraded forestlands and village commons used earlier for free grazing of their cattle) the project will report losses. Even these losses can be contained if the direct benefits from some community woodlots, for which information was not available, and other indirect benefits, mostly of an environmental nature, are included. The findings of this study suggest that watershed development projects initiated to improve the economy and ecology of India's dry and semiarid regions are economically viable and socially desirable. PMID- 11436664 TI - A 400-year tree-ring chronology from the tropical treeline of North America. AB - High-elevation sites in the tropics may be particularly sensitive to rapid climate change. By sampling treeline populations, I have developed the first extensive (> 300 years) tree-ring chronology in tropical North America. The site is Nevado de Colima, at the western end of the Mexican Neovolcanic Belt, and the species studied is Mexican mountain pine (Pinus hartwegii). Despite past logging in the area, 300 to 500-year old pines were found at 3600-3700 m elevation, about 300 m below the present treeline. The Nevado de Colima tree-ring chronology is well replicated from 1600 to 1997. Calibration with Colima climatic records points to summer monsoon precipitation as the strongest dendroclimatic signal. Most trees also exhibit extremely low growth in 1913 and 1914, following the January 1913 Plinian eruption of the Volcan de Colima. Because P. hartwegii is found on top of high mountains from Mexico to Guatemala, there is potential for developing a network of tropical treeline chronologies. PMID- 11436665 TI - [Arterial hypotension in dialysis]. AB - Arterial hypotension, defined as a systolic blood pressure < 100 mmHg, is the most frequent complication in dialysis patients. Four types of hypotension can be identified: chronic, per-dialytic, hypotensive shock and the hypotension due to an unexpected cause. The pathophysiology is relatively well known when the hypotension is secondary to a decrease in the intravascular volume provoked by a sustained ultrafiltration rate during the dialysis session. However, new mechanisms also appear to play an-important role, namely, the dysfunction of the autonomic nervous system and the plasmatic accumulation of vasoactive substances such as adrenomedullin, nitric oxide, and asymmetric dimethyl arginine. Hypotensive episodes can be responsible of acute vascular complications such as myocardial ischemia, ischemic cerebro-vascular accidents, venous thrombosis (retina vein), intestinal ischemia and the aggravation of lower member arteritis. In the long-term, pre-dialysis hypotension has been found associated with an increased mortality rate. The goal of this article is to review the pathophysiological mechanisms involved in the arterial hypotension of dialysis patients, its treatment and the potential preventive measures. PMID- 11436666 TI - [Mixed type amyloidomas, beta2 microglobulin and AL lambda, in a long term hemodialysis patient]. AB - A frequent complication of hemodialysis is deposition of beta 2-microglobulin amyloid in tendons and joints. Soft tissues involvement is exceptional. A 74 year old woman on hemodialysis for 21 years developed two bilateral and symmetric tumours in gluteal regions causing discomfort when sitting. Histology, immunohistochemistry and electron microscopy showed that deposit consisted of predominant beta 2-microglobulin and lambda light chain. This is an unusual localisation of amyloidosis developing late in the course of hemodialysis. It may be perhaps initiated by chronic traumatism and be promoted by the light chain lambda of immunoglobulin. PMID- 11436669 TI - [Course of chronic renal failure epidemiology and prediction of maintenance dialysis needs in France]. AB - Epidemiology of diseases leading to end-stage renal disease (ESRD) in France has greatly changed over the past decades, with the disappearance of type 1 primary membranoproliferative glomerulonephritis, and the increased incidence of both vascular and diabetic nephropathies. The incidence of ESRD is continuously growing, by about 4% per year, with a present rate of more than 100 new patients per million population (pmp) per year. The rise in incidence is mainly observed in older subjects. As a consequence, one may predict a relentless increase in the next years, in parallel with the relentless ageing of the population. The number of ESRD patients on maintenance dialysis is also growing, by nearly 4% per year. The present prevalence is 433 pmp in the Ile de France area, ranging from 268 pmp in subjects aged 15-59 years, to as high as 980 pmp in the population aged 60 years or more. Whereas the number of in-center treated patients has remained quite stable over the past five years, the total number and proportion of out center treated patients is continuously increasing, as expected, thanks to the development of self-care hemodialysis and of peritoneal dialysis. These data may help predict logistic requirements for maintenance dialysis in the next years. PMID- 11436670 TI - [The glomerular permeability factor]. PMID- 11436671 TI - Manual hyperinflation: consistency and modification of the technique by physiotherapists. AB - BACKGROUND AND PURPOSE: The present study aimed to evaluate the consistency with which physiotherapists apply manual hyperinflation to a test lung using the Air Viva-2 or Mapleson-B resuscitation circuit, and their ability to modify the technique as pulmonary characteristics change. METHOD: A quasi-experimental, randomized, repeated-measures design was used to study 16 volunteer physiotherapists performing manual hyperinflation to a test lung simulating three clinical situations. Each subject applied manual hyperinflation to the test lung for each simulation three times in one day using the resuscitation circuit that they would normally use in their clinical practice. Eight subjects used the Air Viva-2 circuit and eight used the Mapleson-B circuit. Measurements of tidal volume (Vp), peak airway pressure (Paw) and fraction of delivered oxygen (FDO2) were recorded during each testing period. Inflation rate and minute volume were calculated. RESULTS: As compliance decreased and airway resistance increased, VT decreased and Paw increased. Of the eight subjects using the Air-Viva-2 circuit, only three subjects delivered greater than 0.80 FDO2. All subjects using the Mapleson-B circuit delivered greater than 0.85 FDO2. CONCLUSIONS: Subjects demonstrated good consistency in the application of manual hyperinflation for all three simulations and modified their technique appropriately as simulated pulmonary characteristics changed. PMID- 11436672 TI - Accuracy of reproducing hand position when using active compared with passive movement. AB - BACKGROUND AND PURPOSE: Evaluating proprioception is relevant to physical rehabilitation because of its significance in motor control. One method of proprioceptive testing involves having subjects either imitate or point at a joint position or movement which was presented via a passive movement. However, as the muscle spindles are subject to central fusimotor control, the proprioceptive system may be better-tuned to movements created by active muscular contraction than to passive movements. The objective of the present study was to determine whether accuracy of reproducing hand position is dependent on whether proprioceptive input is obtained via an active or a passive movement. METHOD: Thirty-nine healthy volunteers (mean age (+/- SD) 24.6 (+/- 3.6) years) participated in the study. Subjects' right hands, which were obscured from view, were acoustically guided to five targets on a digitizer tablet with either an active or passive upper extremity movement. Subjects were then asked to reproduce the targets' location by either reaching to them with the unseen hand or by use of a laser beam. Distance from target and angular deviations were calculated in both absolute and relative terms. Repeated measures analysis of variance (ANOVA) was performed for each variable followed by predetermined contrasts. RESULTS: Comparison between the active and passive conditions when reconstruction of target location was guided kinaesthetically indicates significant differences in absolute distance, range and angular deviation. The comparison when reconstruction of target location was guided visually indicates significant differences in absolute distance, absolute angle and angular deviation. CONCLUSIONS: The ability to reproduce hand position accurately is enhanced when position is encoded by active upper extremity movement compared with passive movement. The results have implications for the design of strategies for evaluating as well as treating patients with impaired proprioception and limited movement. PMID- 11436673 TI - Effect of elbow flexion contractures on the ability of people with C5 and C6 tetraplegia to lift. AB - BACKGROUND AND PURPOSE: It is commonly assumed that minor elbow flexion contractures prevent people with C5-C6 tetraplegia and paralysis of the triceps brachii muscles from bearing full body weight through their upper limbs. The aim of the present study was to determine the effect of simulated bilateral elbow flexion contractures on the ability of these individuals to bear weight through their upper limbs and to determine whether full passive elbow extension is truly critical for lifting body weight. METHOD: A biomechanical study was performed. Body weight lifted was measured under conditions that simulated bilateral elbow flexion contractures. Five people with motor complete C6 tetraplegia and one person with motor complete C5 tetraplegia, all with bilateral paralysis of the triceps brachii muscles, were recruited to the study. Subjects were fitted with bilateral elbow splints that restricted elbow extension but did not restrain elbow flexion nor prevent the elbow from collapsing, and were seated on an instrumented platform that measured vertical forces under the buttocks. Subjects pushed down through their hands and lifted under five different conditions, namely: with no elbow splints; with bilateral elbow splints adjusted to restrict elbow extension by 5-10 degrees; by 15-20 degrees; by 25-30 degrees and with bilateral elbow splints adjusted to allow unrestricted movement of the elbow joint. Maximal weight lifted from under the buttocks, for each condition, was expressed in relation to weight under the buttocks during unsupported sitting (that is, 'seated body weight'). RESULTS: Subjects lifted progressively less weight from under their buttocks as passive elbow restriction was progressively restricted. However, one subject lifted all his seated body weight when elbow extension was restricted by 5-10 degrees and another lifted all his seated body weight when elbow extension was restricted by 5-10 degrees and 15-20 degrees. CONCLUSIONS: Minor elbow flexion contractures will not alone prevent people with tetraplegia and paralysis of the triceps brachii muscles from lifting. Full passive elbow extension is not critical for the performance of this task. PMID- 11436674 TI - Concurrent validity of the NK hand dexterity test. AB - BACKGROUND AND PURPOSE: The present study evaluated the concurrent validity of the NK hand dexterity test (NKHDT) by use of three separate analyses: (1) the correlation between the NKHDT and a criterion comparator (Jebson's Hand Function Test (JHFT)); (2) the correlation between both dexterity tests and a patient rated function questionnaire; and (3) the ability of subscales to differentiate between subjects with and without upper extremity pathology. METHOD: The study population included 40 individuals with a variety of musculoskeletal problems affecting the upper extremity and 10 individuals without any history of upper extremity problems. Both dexterity tests were administered on a single occasion according to a standard protocol. Subjects also completed a rating scale which evaluated self-care, household work, work and recreation on an 0-10-point scale. RESULTS: The validity of the NKHDT was supported in all three analyses because: (1) the correlation between the NKHDT and JHFT subtests was moderate to strong (Pearson's r = 0.47-0.87) and stronger when the objects were more similar in size; (2) both scales correlated to a similar extent with patient-rated function (Pearson's r = 0.34-0.67); and (3) all subscales were statistically different between subjects with and without upper extremity pathology (p < 0.01). CONCLUSIONS: The present study supports the use of the NKHDT as a measure of hand dexterity. PMID- 11436675 TI - Vertical anthropometric measures and low back pain in adolescents. AB - BACKGROUND AND PURPOSE: The present paper explores aspects of a large dataset of adolescent potentiates of low back pain. Standing, trunk and shoulder height, subischial height, weight and mean leg length are examined in relation to reports of low back pain in the last two weeks, in strata of age and gender. METHOD: The present study examined 1254 adolescents, aged 11-18 years, drawn from 12 Adelaide public high schools during 1998. RESULTS: At each year of age, girls reported significantly more low back pain than boys. Girls and boys at each year of age provided different relationships between low back pain and anthropometric features, highlighting the unique nature of adolescent growth. The most consistent findings were anthropometric differences between 16-year-old boys, and 13- and 15-year-old girls, with and without low back pain, who may be at critical stages of the peak growth curve. CONCLUSION: We suggest a high prevalence of low back pain in adolescents, and that young sufferers with low back pain may be reflecting attributes of their stage of anthropometric growth. PMID- 11436676 TI - Progress by design. PMID- 11436677 TI - [The effect of bronchofiberoscopy on selected markers of circulatory system function]. AB - Bronchofiberoscopy is one of the basic diagnostic tools in pulmonary care. It is therefore of interest to assess the effects of this procedure on circulatory system as well as the risk of possible cardiovascular complications. The study included 56 patients for whom bronchoscopy was a planned diagnostic procedure (9 patients had additionally BAL). Each patient's examination protocol included, among others, ECG by Holter method, the measurement of arterial pressure and 12 lead ECG performed before and after the examination. The patients were divided into two groups: those with ventricular arrhythmia diagnosed by ECG at rest and those without arrhythmia signs in ECG done prior to bronchofiberoscopy. The analysis of data revealed that bronchofiberoscopy was associated with considerable increase of arterial pressure in both groups of patients as well as with increased ventricular arrhythmia, particularly in patients with previous history of arrhythmia. Additional load upon the respiratory system exerted by BAL did not have a significant effect on the circulatory system. PMID- 11436678 TI - [The effect of long-term administration of isosorbide 5-mononitrate (Mononit), nitric oxide donor, on gallbladder emptying in patients with chronic stable angina]. AB - In 13 patients (7 females and 6 males) with newly found, previously not treated, chronic stable angina the interprandial volume of the gallbladder and its emptying after treatment with isosorbide-5-mononitrate (Mononit) were examined. The volume of gallbladder was estimated with ultrasonography first after administration of 10 or 20 mg of Mononit twice a day and then in 3rd and 6th week of treatment. It was found that the intake of the single dose as well as the long treatment with Mononit had no effect on interprandial volume of gallbladder. However the intake of drug led to the significant inhibition of postprandial gallbladder's contractility both after the single dose and long term treatment with Mononit. PMID- 11436679 TI - [Leukocytosis in the first day of acute ischemic stroke as a prognostic factor of disease progression]. AB - It has been reported that white blood cells (WBC) may contribute to the initiation and further development of stroke. WBC count could have influence on hemorheology, thrombosis and induction of vasoconstriction in cerebral arteries. The aim of this work was to determine whether WBC count assessed in patients with acute stroke in the first day of the disease have a predictive value for the late clinical outcomes. Peripheral WBC count was measured at the admission to the hospital in 100 consecutive patients with acute stroke. Ischemic stroke was confirmed in all patients by head CT. Neurological status was evaluated with the use of modified Rankin's Scale. We assessed the following clinical outcomes: in hospital mortality, the lack of neurological improvement on the 21st day of disease, and the need for hospital stay longer than 21 days. Odds ratio (OR) with 95% confidence interval (95% CI) adjusted for patient's age, gender, cardiovascular diseases, diabetes mellitus and previous stroke was calculated with the use of logistic regression. OR was evaluated both for the presence of WBC count > 10.000 cells/microliter and for each 1000 leukocytes/microliter increase. WBC count greater than 10.000 cells/microliter was associated with a more than seventy-fold greater risk for death (OR--75.18; 95% CI: 8.89-635.84; p = 0.0001), nearly eight times increased risk for the no-improvement status (OR- 7.78; 95% CI: 2.51-24.17; p = 0.0004) and approximately four times increased risk for the prolonged hospital stay (OR--4.20; 95% CI: 1.50-11.72; p = 0.0062). Each 1000 cell/microliter increase in WBC count at the admission was associated with increased risk for in-hospital mortality (OR--2.24; 95% CI: 1.39-3.57; p = 0.0008), no neurological improvement (OR--1.43; 95% CI: 1.15-1.76; p = 0.0009) and the need for prolonged hospitalization (OR--1.26; 95% CI: 1.07-1.49; p = 0.0066). Increased WBC count within the first 12 hours of stroke is an independent and strong risk factor for mortality, no neurological improvement, and the need for prolonged hospitalization in the course of acute ischemic stroke. PMID- 11436680 TI - [Usefulness of thoracic imaging and computed tomography in evaluation of neoplastic changes of lungs and mediastinum in children]. AB - Radiological methods of imaging diagnostics allow to evaluate exactly and to monitor the treatment course of pathological lesions in chest. Basic examinations are: plain chest X-ray and computer tomography. Optimal diagnostic algorithm of neoplastic changes in chest is not always univocally defined. The aim of the study is to compare the results of estimation of the presence and type of neoplastic changes in mediastinum and lungs based on X-ray and computer tomography. The results indicate that initial and control X-ray examination allows to diagnose mediastinal lymphadenopathy coexisting with pulmonary hilus extension. CT is used to diagnose and monitor lung tissue and mediastinal changes. PMID- 11436681 TI - [The role of cell-mediated immune response in pathogenesis of alopecia areata]. AB - Recent studies have suggested that cell-mediated immune response play a critical role in the pathogenesis of alopecia areata (AA). Eighteen patients with AA were included in the study. Fifteen healthy subjects served as controls. Serum levels of sTNF alpha RI and sIL-2R were measured using enzyme-linked immunosorbent assay technique. The serum levels of sTNF alpha RI were significantly elevated in patients with AA in comparison with control group. The serum levels of sIL-2R were higher in AA patients than in healthy subjects but not significantly. These results indicate, that immune mechanisms in AA are characterized by activation of T cells and other cells, possibly keratinocytes. PMID- 11436682 TI - [Evaluation of urinary tract function in various types of urinary incontinence in women]. AB - 164 women from 18 to 74 years of age (mean 48.3) with urinary incontinence, previously not operated, with out any neurogenic symptoms, with sterile urine were studied. The kind of incontinence was estimated on the basis of clinical history and examination, the analysis of frequency/volume charts, free uroflowmetry and cystometry in sitting and standing positions. Genuine stress incontinence (GSI) was recognized in 121 (73.8%) women, urge incontinence (UI) in 15 (9.1%), and mixed incontinence (MI) in remaining 28 (17.1%) women. Women with UI and MI had significantly more often both daytime (12.5 +/- 6.4 and 10.3 +/- 2.5), and night-time mictions, comparing to GSI (7.2 +/- 2.8) with simultaneously smaller daytime voided volumes (91 +/- 36.0 ml and 125 +/- 50.0 ml) comparing to GSI (175 +/- 151 ml). Mean voided volume and Qmax in free uroflowmetry were significantly greater in GSI (293 +/- 124 ml, 24.6 +/- 8.4 ml/s comparing to 165 +/- 69.0 ml, 17.0 +/- 7.8 ml/s in UI). The bladders of women with UI showed motoric and sensoric hyperactivity, uninhibited detrusor contractions, decreased CysCapmax, higher PdetCysCapmax, and decreased compliance. Mean CysCapmax was 240 +/- 102 ml, with first sensation after filling mean 145 +/- 90.1 ml, with Pdet 29.3 +/- 13.4 cm H2O, and compliance 13.2 +/- 7.7 ml/cm H2O. In women with both GSI and MI the values were significantly higher and were respectively, for CysCapmax 455 +/- 148 ml, 308 +/- 58 ml, for VFD 237 +/- 76 ml and 169 +/- 25 ml, PdetCysCapmax 12.9 +/- 5.4 cm H2O and 16.5 +/- 4.9 cm H2O, and for C 41.7 +/- 20.5 ml/cm H2O and 28.2 +/- 8.1 ml/cm H2O. The urge syndrome accompanied by urine incontinence, decreased functional bladder capacity evaluated from frequency/volume charts and free uroflowmetry are the indications for performing cystometry with simultaneous monitoring of abdominal pressure. It allows to recognize urge and mixed incontinence. PMID- 11436683 TI - [Levels of soluble intracellular adhesion molecules 1 (sICAM-1) , soluble receptors for interleukin 2 (sIL-2R) and anti-Epstein-Barr viral capsid antigen (anti-VCA IgM) in blood serum during the course of infectious mononucleosis]. AB - The clinical picture of infectious mononucleosis and the consequences of EBV infection are due to immune response mechanisms. The level of soluble form of IL 2 receptor (sIL-2R) is thought to be a marker of T-cell activity, especially CD8+. Intercellular adhesion molecule (ICAM-1, CD 54) plays an important role in the process of antigen presentation. The aim of this study was to assess the serum concentration of sIL-2R, ICAM-1 and anti-VCA IgM in patients with infectious mononucleosis. The study group comprised 42 persons: 20 healthy subjects as a control group and 22 individuals with infectious mononucleosis. The highest anti-VCA IgM serum level in all patients was during the 1st day of hospitalization, and decreased in the 8th day of hospitalization. The lowest antibody concentration was observed when the symptoms and sings ceased. The level of sIL-2R was significantly increased and during farther hospitalization we observed lower, but still elevated concentrations. Our study has demonstrated statistically significant elevation of sICAM-1 level during the entire period of hospitalization. This data indicates the importance of antigen presentation process. Although the serum concentration of immune response mediators does not reflect their contents in organism, but is a useful method for in vivo examination. PMID- 11436684 TI - [Chronic hepatic encephalopathy--clinical features and diagnosis]. AB - Neuropsychiatric abnormalities affecting patients with chronic liver disease are termed chronic hepatic encephalopathy. The spectrum of hepatic encephalopathy varies from mild intellectual impairment to deep coma. The diagnosis is based on clinical findings, and psychometric and electroencephalographic testing. PMID- 11436685 TI - [Hepatitis C virus infection (HCV) as a pediatric problem]. AB - Hepatitis C is a major health problem also in children. General screening is not advisable. Screening should be limited to the risk groups. Nowadays there are not any large studies on the treatment of chronic hepatitis C in children. Available studies suggest that children have response rates to interferon monotherapy similar to adults. PMID- 11436686 TI - [Non-insulin dependent diabetes mellitus during development in the population ]. AB - This type of diabetes mellitus is generally considered rare in children and adolescents. The incidence of type 2 diabetes has increased dramatically in the past decade in some ethnic groups. This increased incidence of youth-onset non insulin dependent diabetes mellitus (NIDDM-Y) has corresponded temporally to an unprecedented increase in body weight and prevalence of obesity in youth in various ethnic populations. Maturity-onset diabetes of the young (MODY), another type of non-insulin-dependent diabetes mellitus, is characterized by an early onset and autosomal dominant inheritance. MODY blood glucose is primarily due to beta-cell defects resulting in inadequate insulin secretion for a given level. In contrast to NIDDM-Y there is no significant increase in insulin resistance. PMID- 11436687 TI - [The role of selenium in skin]. AB - The paper reviews the data on the content of selenium in the skin and its appendages and the influence of the micronutrient on skin functions. The paper focuses on antiproliferative, photoprotective and immunomodulatory properties of selenium compounds. Dermatological symptoms due to selenosis are described and skin disorders with low selenium status are mentioned. PMID- 11436688 TI - [The circulatory system in children suffering from progressive muscular dystrophy]. AB - The aim of the study was to present the state of circulatory system in course of progressive muscular dystrophy. The results of clinical examination, as well as electrocardiography, echocardiography, cardiac catheterization, chest x-ray and histopathology of the tissues of the chest and vessels were described. PMID- 11436689 TI - [Kawasaki disease--personal observations]. AB - The aim of the study was to analyse two cases of Kawasaki disease--the first one with myocardial infarction and the second with positive outcome. Kawasaki disease is a fever disease usually occurring in very young children. Vasculitis is a characteristic feature of Kawasaki disease. Etiology of this disease is unknown. Diagnosis is based on typical clinical symptoms such as: fever, rash, conjunctivitis, lymphadenitis colli, inflammation of the mucous membranes, erythema of the hands and feet. The treatment can prevent thrombosis, myocardial infarction which are the most serious complications of the disease. Kawasaki disease is treated with intravenous gamma--globulin therapy and high dosage acetylsalicylic acid at the beginning of the disease. Such a treatment of Kawasaki disease prevents the coronary vessel lesions. PMID- 11436690 TI - [Potter sequence in a neonate with bilateral renal dysplasia with genetic aspects]. AB - The authors describe a neonate with Potter sequence due to bilateral renal dysplasia and discuss its genetic implications. Although this congenital anomaly of the kidneys may occur sporadically, it is now accepted that this condition has an autosomal dominant inheritance pattern with a penetrance of 50-90% and variable expression. Potter sequence is not compatible with extrauterine life; therefore, in case of maternal oligohydramnios, ultrasound should be performed in the second trimester in order to make a prenatal diagnosis, as well as to permit genetic counseling. As Potter sequence is frequently associated with clinically silent anomalies of the kidneys, ultrasound examination of the kidneys and urinary tract should also be performed in all family members. PMID- 11436691 TI - [Sacral bone dysgenesis in a child whose mother has diabetes]. AB - We introduced 2.5-year old girl with gait and miction disturbances as a result of sacral bone dysgenesis (only S1 existed) and abnormal position of nerve roots of medulla. The 28-year old mother of the child has been treated for diabetes mellitus type I since she was 15. PMID- 11436692 TI - [Widened resection in locally advanced carcinomas of the right half of the colon]. AB - We describe two cases of locally advanced carcinomas of the right half of colon. In spite of big tumour size and infiltration of adjacent organs we decided to perform widened "en block" resection. Microscopically no neoplastic cells were found in lymph nodes. During a year observation we did not ascertain any features of neoplasm relapse what is consistent with data published by other authors. They indicate that radical resection of big-sized tumours infiltrating adjacent organs in the case of absence of metastases into lymph nodes gives the patient a chance for long-term survival. PMID- 11436694 TI - Getting well on the road. PMID- 11436693 TI - [Surgical treatment of colonic diverticulitis--personal observations]. AB - The aim of paper is presentation of our experience in surgical treatment of diverticular disease. In the period of 11 years we operated on 22 patients aged from 38 to 85 years because of complicated diverticulitis. We admitted to hospital 11 patients as emergency cases; operation was performed urgently in 9 of them due to symptoms and signs of acute peritonitis. The other patients were admitted to hospital because of stenosing inflammatory pericolic mass, pericolic abscess or at least two episodes of acute diverticulitis in the past. Resection of affected part of colon was performed in all patients. Primary anastomosis was performed in 13 patients, Hartmann's operation was performed in 8 patients and suture of caecum in site of resected diverticulum was performed in one patient. There was no mortality. Perioperative complications occurred in 31.8% of patients; anastomotic leak occurred in 1 (7.7%) of 13 patients with primary bowel anastomosis. Results were discussed on the base of recent literature. PMID- 11436695 TI - [Outbreak of Dengue-2 virus in Salta, Argentina, 1988]. AB - Dengue (DEN) reemergence was first detected in Salta, Argentina, in 1997. It was confirmed by serology and PCR that DEN-2 virus was responsible for sporadic cases and indigenous transmission. A laboratory-based surveillance system allowed the detection of an outbreak in Salta several months later. In total, 378 DEN cases were laboratory diagnosed out of 646 (58%) studied. The cases were distributed in 10 localities of the province between January 3rd and May 31st, 1998. One DEN-2 viral isolation and 4 PCR products were obtained from patients coming from Oran and Embarcacion localities. Male and female cases occurred at a similar rate (1:1), with adult cases (82.5%) nearly five times greater than for children under 15 years-old (17.5%). Clinically, all cases corresponded to classic DEN. This is the first time that a DEN outbreak has been laboratory-diagnosed in Argentina and also the first time that DEN virus has been isolated in this country. PMID- 11436696 TI - [Evaluation and cost of the health care process of diabetic patients]. AB - The aim of this randomized trial o evaluate the quality and cost of health care provided to people with diabetes in a Public Employment-Based Health Insurance program in La Plata, Argentina. Consequently, 1590 diabetic patients were interviewed and the data obtained were analyzed using univariate and multiple logistic regression. Patients treated without insulin (48%) were significantly older and reported obesity, hypertension, and macroangiopathy more frequently than those treated with insulin. Hospitalization rates were significantly higher in insulin-treated patients. These rates were comparable in people without complications or microangiopathy alone, but increased significantly in patients with macroangiopathy. Many recommended practices were not performed during the preceding year: foot exams (20-30%), referral to ophthalmologist (21-29%), and HbA1c test (60-78%). In contrast, the rate of medical testing and visits was high, even in uncomplicated patients, resulting in an estimated cost of the care 22.7% higher than it would have been if the tests and procedures had been performed as recommended by international guidelines. We conclude that the current care for diabetes is inefficient and expensive; the implementation of data-based guidelines would result in a more effective allocation of resources, thus improving the quality of diabetes care and overall quality of life. PMID- 11436697 TI - [Molecular variants of human papillomavirus (HPV) types 16 and 18 in adenocarcinomas of the cervix]. AB - Human Papillomavirus (HPV), placebo clinical trial particularly types 16 and 18, are considered human carcinogens since an etiological association has been demonstrated between these viruses and the development of cervical cancer. While the viral role in squamous carcinoma has been largely studied, the information available on adenocarcinoma is scarce, partly because of its lower frequency. In this paper we investigated the presence of HPV types and intratype variants in adenocarcinomas of the cervix. A total of 23 archive samples, fixed and paraffin embedded biopsies, were included. The detection and viral typing was performed by generic PCR and subsequent single stranded conformational polymorphism analysis (SSCP). Genetic variability was investigated in a 450 bp-fragment corresponding to L1 gene by post-PCR direct sequencing. We detected 11 HPV 16 positive samples (9 prototypes and 2 variants: 1 European and 1 Asiatic-American), 10 HPV 18 (9 prototypes and 1 European variant), 1 HPV 31 and 1 negative. The high risk HPV association with this neoplasia was confirmed with a high prevalence (43%) of HPV 18, (but) without predominance over the other types as previously published. The demonstrated variability in L1 protein epitopes originated aminoacidic changes which could have implications on the immune response and therefore should be considered in a vaccine design. PMID- 11436698 TI - [Ha-ras and p53 gene mutations scanned by PCR-SSCP in premalignant and malignant lesions of the uterine cervix associated with human papillomavirus]. AB - The aim of this study w trial randomized as to investigate the frequencies of human papillomavirus (HPV) and mutation in Ha-ras oncogene and tumour suppressor p53 gene in cervical cancer and precursor lesions. A total of 30 invasive carcinomas (IC), 36 cervical intraepithelial neoplasia grade III (CIN III) and 12 normal tissues adjacent to the tumor (NT) were included. HPV typification and scanning of possible mutations in Ha-ras and p 53 genes were made by SSCP-PCR. The IC cases showed 93% HPV positivity, 41% having mobility shifts for Ha-ras mutations and 17% for p53 mutations while in CIN III, these percentages were 80%, 18% and 11%, respectively. In normal tissues HPV frequency was 17%. All Ha-ras mutated samples were HPV positive but 33% of p53 mutated cases were HPV negative. All mutations were heterozygous. HPV 16 was more prevalent (44%) than HPV 18 (15%) and the high rate of undetermined HPV types (18%) would indicate the circulation in our country of other types different from the assayed HPV controls (6, 11, 16, 18, 31 and 33), being variants or mixed infections. The low frequency of p53 mutations (17%) strengthens the view that wild type p53 inactivation by HPV probably plays a major role in the pathogenesis of cervical cancer. Because mutated Ha-ras was found in HPV associated premalignant lesions, we speculate that it represents an early marker for progression. Our findings provide additional evidence for an interactive effect between high risk types of HPV and oncogene activation in the development of uterine cervical cancer. PMID- 11436699 TI - [Epworth drowsiness scale value in obstructive sleep apnea syndrome]. AB - Hypersomnia is one of the most consulted symptoms among patients evaluated at sleep disorder centers and it is frequently related to obstructive sleep apnea syndrome (OSAS). Our hypothesis is that Epworth sleepiness scale (ESS) is the parameter with the greatest predictive value in the OSAS diagnosis. We compared patients with OSAS diagnosis to a control group. In both groups we compared ESS with body mass index (BMI), neck circumference (NC), waist perimeter (WP). Anthropometric index (BMI, NC and WC), were similar in both groups (p < 0.10). When we analyzed ESS, a score greater than 10 was observed in the OSAS group, with a significant difference between groups (p < 0.001). Epworth sleepiness scale yielded 60% of sensibility, 82% of specificity and a positive predictive value of 85%. The negative predictive value was 52%. Confidence index was 70%. The relationship between OSAS and ESS scale was significant (Pearson Chi-Square value 7.5). Odds Ratio for apneas was 15 and its confidence interval was lower than 1.5 and upper than 141. We conclude that with ESS score exceeding 10 points OSAS should be suspected. PMID- 11436700 TI - [Evaluation and treatment of interstitial lung disease in Argentine]. AB - To determine the availability and usual management of interstitial lung diseases (ILD) in our country, the Section of Interstitial Lung Diseases of the Argentine Association for Respiratory Medicine (AAMR) made a survey about diagnostic methodology and treatment of ILD. A total of 115 answers were obtained (38.5%), 43% of them among physicians living in the provinces. Availability of diffusing capacity of the lung for carbon monoxide test (DLCO) is limited: 25.4% never have access to it and 35.6% can seldom use it. Availability to thoracic CT scan is wider: 85% may use if often (32.4%) or always (52.6%). Bronchoscopy is commonly available in 87.7% of the physicians either often (21.9%) or always (65.8%). However, only 20.2% perform BAL and 13.1% transbronchial biopsy in every patient. Only 16.6% perform open lung biopsy or thoracoscopic biopsy in all or most of their patients. Sixty eight percent of physicians who always have availability of DLCO perform it in every patient but only 7.1% of those who seldom have access to DLCO do so (p = 0.0003). Availability of bronchoscopy does not have any influence on the decision of performing BAL or transbronchial biopsy. Frequency of use of surgical biopsy or treatment with immunosuppressive drugs was not influenced by any variable. We conclude that there is a current trend to underuse diagnostic resources for ILD in Argentina. Limitations in availability are relevant regarding DLCO. An effort from the health authorities to centralize the management of patients with ILD would allow to study and treat them according to international recommendations. PMID- 11436701 TI - [Pharmacokinetic variability of oxcarbazepine in epileptic patients]. AB - The aim of this study was to analyze the population pharmacokinetics of oxcarbazepine (OCBZ) measuring the serum level of its active metabolite, monohydroxylated oxcarbazepine (MHD). We studied a group of patients with symptomatic and cryptogenic epilepsy treated with OCBZ monotherapy orally, at least for 3 weeks. The mean doses, age and weight of the patients were 17.9 +/- 7.8 mg/kg/day, 35.6 +/- 16.4 years and 70.3 +/- 19.2 kg, respectively. Blood samples were taken before the first morning dose of OCBZ and MHD levels were determined by HPLC. A linear relationship was found between OCBZ dose and MHD serum level (r = 0.844, p < 0.001). The MHD serum concentration (mg/l) can be predicted as 0.85 x OCBZ dose (mg/kg). There was a significant correlation between observed and predicted MHD concentrations for each patient. The mean MHD clearance (Cl/F) calculated was 4.05 +/- 1.69 l/h, with a coefficient variation of 41%. It was independent of dose, age and weight and followed a non normal distribution. The half-life of MHD was 10.50 +/- 3.17 hours. The influence of other antiepileptic drugs on MHD pharmacokinetics was analyzed by comparing the Cl/F medians from groups of patients receiving concomitant drugs with OCBZ monotherapy group where no significant differences were found. The results can be used to estimate a priori OCBZ doses, in order to individualize the treatment. PMID- 11436702 TI - [Low prevalence of autoimmune antiphospholipid antibodies in hepatic diseases]. AB - Antiphospholipid antibodies (aPL) have been associated with different diseases. They are defined as a large family of immunoglobulins (Ig) of either alloantibodies or autoantibodies. The autoimmune antibodies are associated with venous and/or arterial thrombosis, thrombocytopenia and recurrent fetal loss in the so-called antiphospholipid syndrome or in systemic lupus erythematosus. These antibodies are directed against proteins or phospholipid-protein complexes. On the contrary, antiphospholipid antibodies (alloantibodies) which are found in infectious diseases sera (syphilis, HIV, and other viral diseases), disappear with illness remission and are directed to phospholipids alone (particularly cardiolipin) and are not associated with thrombosis or recurrent fetal loss. However, the role and type of aPL found during hepatic diseases is still unclear. To investigate the prevalence of autoimmune aPL (IgG and IgM) during different hepatic diseases, we have studied 128 patients with hepatitis C virus, hepatitis B virus and hepatic autoimmune diseases without treatment as well as 40 healthy control subjects. We have used a specific ELISA kit, that uses a mixture of phospholipid instead of cardiolipin alone, and allows a better detection of aPL of the autoimmune type. Our results show that autoimmune aPL are not significantly increased in viral hepatic diseases (2%) or autoimmune diseases of the liver (3%) when compared to the control group (0%). PMID- 11436703 TI - [Effects of competitive substrates ans insulin on glucose uptake and utilization in isolated perfused hearts of dyslipemic rats]. AB - Rats chronically fed (15 weeks) a sucrose-rich diet (SRD) developed hypertriglyceridemia (hyperTg), increased plasma free fatty acids (FFA), impaired glucose homeostasis and insulin insensitivity. An increase of Tg and glycogen (Gly) in heart muscle was also observed. HyperTg with altered glucose metabolism could have profound effects on myocardial glucose utilization. To test this hypothesis male Wistar rats were fed a semi-synthetic SRD (w/w: 62.5% sucrose, 8% corn-oil, 17% protein), and the control group (CD) received the same semi synthetic diet, except that sucrose was replaced with starch for 90 days. At that time, the hearts from these animals were isolated and perfused for 30 min in the presence or absence of insulin (30 mU/ml). Levels of the exogenous substrates were similar to those found in the plasma of the animal in vivo in both dietary groups (glucose 8.5 mM, palmitate 0.8 mM in SRD and glucose 5-5 mM, palmitate 0.3 mM in CD). In the absence of insulin glucose uptake was reduced (40%) and lactate release was increased (50%) in SRD hearts. Glucose oxidation was depressed mainly due to both, an increase of PDH kinase and a decrease of 60% of PDHa (active form of PDHc). Insulin in the perfusion medium improved only glucose uptake. The results suggest that at least two different mechanisms might contribute to insulin resistance and to impaired glucose metabolism in the perfused hearts of dyslipemic SRD fed rats: 1) reduced basal and insulin-stimulated glucose uptake and its utilization and 2) increased availability and oxidation of lipids (low PDHa and PDH kinase activities), which in turn decreased glucose uptake and utilization. Thus, this experimental model may be useful to study how impaired glucose homeostasis, increased plasma FFA and hyperTg could contribute to heart tissue malfunction. PMID- 11436704 TI - [Differential binding of hyaluronic acid in 2 CD44+ sublines. Relationship with tumor infiltration]. AB - We have established and characterized a cell line (LBL) from a spontaneous murine T lymphoma LB. Histopathological analysis has demonstrated LB primary tumor infiltration in spleen, lymph nodes, liver, thymus, bone marrow and lung. However LBL cells infiltrated all these organs except lung. Two sublines with different growth behavior were derived from LBL cell line. One of them grew in suspension as clusters (LBLc) while the other one grew as adherent monolayers (LBLa). Growth rate, response to mitogenic stimuli and apoptosis induction were different among the parental cell line and the derived sublines. CD44 was expressed constitutively in LBL and LBLa cells. In contrast LBLc cells only expressed similar levels of this molecule when stimulated with PMA. LBLa cells showed hyaluronic acid (HA) binding properties, while LBL and LBLc cells were not able to bind HA even when activated with PMA. We postulate that differences in HA binding could be related with different infiltration behaviors. PMID- 11436705 TI - [Isoforms modulation of CD44 adhesion molecule in a murine model of ischemia and intestinal reperfusion]. AB - Gut ischemia-reperfusion (G-IR) induces a systemic inflammatory response, in which leukocyte contribution to this injury in distant organs is important. ICAM 1 as well as CD11/CD18 have been involved in leukocyte infiltration in liver and lungs. CD44 adhesion molecule plays an essential role in other inflammatory processes such as rheumatoid arthritis and allergic contact dermatitis, however its implication in G-IR has not been described. In order to establish a possible role of CD44 in the development of systemic inflammation by G-IR, we have studied CD44 mRNA expression by RT-PCR in a murine model of gut ischemia reperfusion. Animals subjected to G-IR showed an increased number of CD44 variable isoforms expressed in liver and spleen compared to non-treated animals or animals subjected to laparotomy. This finding indicates that G-IR specifically induces the expression of different CD44 variable isoforms. Liver CD44 upregulation in animals subjected to G-IR suggests a contribution of this molecule to lymphocyte activation and migration to this injured organ. Moreover, increased isoform expression in spleen may be induced by the proinflammatory environment resulting from a systemic depuration activity. PMID- 11436706 TI - [Kikuchi-Fujimoto necrotizing lymphadenitis. Report of 2 cases]. AB - Kikuchi-Fujimoto disease (KFD) is a form of necrotizing lymphadenitis. The disease is a usually benign, self limited cause of lymph node enlargement affecting predominantly young women. Several other clinical manifestations have been described, though the only consistent findings are a macular eruption and a fever. Occasionally, the disease has presented as a fever of unknown origin. Lymph node specimens show areas of necrosis without neutrophilic infiltration. Lymphocyte mediated apoptotic mechanisms are responsible for the necrosis observed. Clinically, it should be differentiated from many different conditions, including infections such as cat scratch disease and rheumatological diseases like Still's disease and Systemic Lupus Erythematosus. Pathologically, it should be differentiated from lymphomas. KFD is widely known in the Far East, but very rare in Latin America. As a matter of fact, we have only detected four reported cases from Argentina, Brazil and Chile. We here describe two additional cases from Argentina. Both occurred in young women. One had a mediastinal compromise and a recurrent course, only observed in 4% of cases. The second one had a classical cervical presentation. The diagnosis in both was made by lymph node biopsy, although, in areas of great prevalence, an aspiration may be sufficient to establish the disease. We anticipate that the real prevalence of this unusual disease is highly underrepresented. PMID- 11436707 TI - [Primary aortoenteric fistula]. AB - Aortoenteric fistulas are a very unlikely cause of digestive bleeding. We present the case of a fifty six year old patient with a primary aortoduodenal fistula who consulted because of melena and a pulsating abdominal mass. He underwent three digestive endoscopies without arriving at any diagnosis. A computed axial tomography and an angiography showed an aneurysm of the abdominal aorta. With an exploring laparotomy a fistula was identified between the aneurysm and the fourth portion of the duodenum. An aortoaortic bypass was carried out followed by a surgical fix of the duodenum. The patient was released six days after the operation. PMID- 11436708 TI - [Streptococcal infection, acute kidney failure and interstitial nephritis]. AB - The relationship between streptococcal infection and renal disease has been object of multiple studies. Streptococcal infection may induce acute glomerulonephritis or interstitial nephritis. We report a patient with a streptococcal infection who developed acute renal failure. The renal biopsy showed an acute interstitial nephritis, with an interstitium infiltrate with a significant number of eosinophils. We review the causes of acute renal failure associated with streptococcal infection, specially acute interstitial nephritis. PMID- 11436709 TI - [Severe dilatation of the pulmonary artery and extrinsic compression of the left coronary arterial trunk in primary pulmonary hypertension]. PMID- 11436710 TI - [Non ketotic hyperosmolar coma, cellulitis and staphylococcal septicemia]. PMID- 11436711 TI - [Accidents due to lepidoptera with special reference to Lonomia sp]. AB - Human poisoning by Lepidoptera (butterflies, caterpillars and moths) has been known to physicians since old times. In the last few years, several factors have been causative of a great number of contacts between different stages of these insects and humans, occasionally with epidemic features. Species of medical interest in Argentina are those related to Megalopygidae, Saturniidae, Hemileucidae, Arctiidae, Notodentidae and Eucliedae families. Among all the species involved, it is important to describe the pathology produced by contact with Lonomia obliqua caterpillar (Saturniidae). The caterpillar Lonomia has several enzymes with procoagulant and fibrinolytic activities, acting on various components of the hemostatic system inducing hemorrhage that can lead to death. In the South of Brazil during the last years caterpillars of this butterfly caused a great number of cases of erucism including some deaths. In the past two years, there have been some cases of poisoning by Lonomia obliqua in Misiones, in the North East of Argentina on the border with Southern Brazil. This presents a potential risk, taking into account the poor knowledge regarding this insect and its venom in Argentina. This brief actualization gives some biochemical, clinical and epidemiologic tools towards understanding human Lepidopterism, an uncommon medical problem in this country. PMID- 11436712 TI - [Osteoporosis in pregnancy and lactation]. AB - Normal pregnancy and lactation lead to a combination of adaptive metabolic responses, the end result of which is to assure adequate delivery of mineral to the fetus while affording protection to the maternal skeleton. Elevated circulating levels of 1.25-OH vitamin D lead to increased efficiency of maternal intestinal calcium absorption and possibly lead to hypersecretion of calcitonin. Although serum concentrations of parathyroid hormone do not change during pregnancy, increased levels of a related hormone, PTH-related peptide, seem to contribute to a state of maternal functional hyperparathyroidism and maintain the fetal-maternal calcium gradient necessary to provide calcium to the fetus. Bone turnover increases during lactation and diminishes urinary calcium loss mobilizing mineral for the milk. Elevated levels of ionized calcium and phosphorus possibly correlate with increased bone resorption and decreased urinary excretion of these minerals. Bone mass is not normally lost during pregnancy but may decrease with sustained lactation for as long as six months. If lactation ceases before nine months, however, bone mass usually is restored. However, several rare forms of osteoporosis have been associated with pregnancy and lactation. PMID- 11436713 TI - [Einthoven cord electrocardiogram]. PMID- 11436714 TI - [Conscience objection]. PMID- 11436715 TI - [Death, veracity, care and family (from Toistoi and Schubert to Kubler-Ross)]. PMID- 11436716 TI - [Nobel Prize of physiology and medicine 2000. Signalling mechanisms in the central nervous system]. PMID- 11436717 TI - No follow-up for indigent patients? PMID- 11436718 TI - Peer review gone awry. The bittersweet victory of Dr. Schulze. AB - This Texas GP endured a two-year inquisition by an HMO and its medical director, then termination and the loss of hundreds of patients. He fought hard to regain his reputation. PMID- 11436719 TI - Know Medicare's rules on screening exams. PMID- 11436720 TI - Online prescribing. How one doctor got caught. PMID- 11436721 TI - Online prescribing. The growing problem of online pharmacies. AB - Attracted by easy money, doctors are prescribing drugs over the Internet for patients they've never met. They could be exposing themselves to professional discipline and malpractice suits. PMID- 11436722 TI - Problem patient? Use the three Ps. PMID- 11436723 TI - A little bit of medicine, a lot of hope. AB - After nearly a week of treating patients in Nicaragua, this physician realized that curing isn't everything. Her story was runner-up for the Young Doctor Award in the Medical Economics Doctors' Writing Contest for 2000. PMID- 11436724 TI - Doctors who go the extra mile. Teaching America's children about AIDS. PMID- 11436725 TI - Making part-time practice work--for all concerned. AB - Doctors are embracing reduced schedules. But issues such as call, overhead, productivity, and partnership remain. PMID- 11436726 TI - Survey offers PMPM benchmarks for selected specialties. PMID- 11436727 TI - Gauge your PMPM costs against these California benchmarks. PMID- 11436728 TI - Focus on adverse drug events to reduce LOS, slash costs. PMID- 11436729 TI - Does higher HMO penetration reduce commercial premiums? PMID- 11436730 TI - Integrated systems still failing to deliver profits. PMID- 11436731 TI - Objects and their locations in exogenous cuing. AB - Anatomical, physiological, and behavioral studies provide support for separate object- and location-based components of visual attention. Although studies of object-based components have usually involved voluntary attention, more recent evidence has suggested that objects may play an independent role in reflexive exogenous orienting, at least at long stimulus onset asynchronies (SOAs). In the present experiments, the role of objects in reflexive attentional orienting was investigated by developing a task in which location and object cuing could be separately examined for both short and long SOAs. Typical location cuing effects were obtained, indicating facilitation at short cue-target intervals and inhibition of return (IOR) at longer intervals. In contrast, object cuing resulted in facilitation for cued objects at long cue-target intervals and no object-based IOR. Interestingly, object cuing primarily affected targets at cued locations, and not those at uncued locations. Together, the experiments examine the interactive nature of objects and locations in exogenous orienting and seem most consistent with a location-mediated view of object-based orienting. PMID- 11436732 TI - Visual attention and the apprehension of spatial relations: the case of depth. AB - Several studies have shown that targets defined on the basis of the spatial relations between objects yield highly inefficient visual search performance (e.g., Logan, 1994; Palmer, 1994), suggesting that the apprehension of spatial relations may require the selective allocation of attention within the scene. In the present study, we tested the hypothesis that depth relations might be different in this regard and might support efficient visual search. This hypothesis was based, in part, on the fact that many perceptual organization processes that are believed to occur early and in parallel, such as figure-ground segregation and perceptual completion, seem to depend on the assignment of depth relations. Despite this, however, using increasingly salient cues to depth (Experiments 2-4) and including a separate test of the sufficiency of the most salient depth cue used (Experiment 5), no evidence was found to indicate that search for a target defined by depth relations is any different than search for a target defined by other types of spatial relations, with regard to efficiency of search. These findings are discussed within the context of the larger literature on early processing of three-dimensional characteristics of visual scenes. PMID- 11436733 TI - Influence of a stereo surface on the perceived tilt of a monocular line. AB - This study addressed the issue of how a stereo-specified surface influences the perceived two-dimensional orientation of a monocularly viewed line. In a series of three experiments, it was found that, in general, when a monocular line was embedded in a disparity field specifying a planar surface slanted about the horizontal axis, the tilt of the line depended on the slant of the surface in which it was embedded and on whether the line was presented in the left or in the right visual field. These results, predicted by perspective geometry, are compatible with the hypothesis that the monocular line is perceived as part of the stereo surface. Moreover, it was found that timing is a crucial factor in determining the strength of the effect. The effect was at a maximum when the monocular line and the stereo surface were presented together, with no prior presentation of the surface. The influence of the stereo surface on the monocular line was substantially reduced, however, when the surface alone preceded the combined presentation of the line and the surface. PMID- 11436734 TI - Perceptual interactions of facial dimensions in speeded classification and identification. AB - The representation underlying the identification and classification of semirealistic line drawings taken from a computer model of the face was investigated by using a speeded classification task and an identification task. These data were analyzed by using a multidimensional extension of signal detection theory, within which varieties of perceptual interactions between dimensions within and across stimuli can be characterized. The dimensions of interest here were eye separation, nose length, and mouth width. The response time and accuracy data from the speeded classification task suggest that processing of a given feature did depend on whether other features were present or absent, but given that other features were present, the results strongly support separability (a macrolevel, across-stimulus form of invariance) for all pairs of facial dimensions used. This separability was confirmed by the subsequent identification task. Owing to its greater resolution, the identification task can reveal interactions that might exist at more microlevels of processing. In fact, the identification data did indicate the presence of perceptual dependence between facial dimensions within a stimulus when the dimensions that were varied were close in spatial proximity (i.e., eye separation and nose length). Within the theoretical framework, perceptual dependence can be interpreted as correlated noise between otherwise separate channels (and hence, is logically distinct from separability). This dependence was greatly reduced for dimensions that were more distant (eyes and mouth). The relation between these results and the configural effects that have been observed with faces as stimuli in other studies is discussed. PMID- 11436735 TI - The ventriloquist effect does not depend on the direction of automatic visual attention. AB - Previously, we showed that the visual bias of auditory sound location, or ventriloquism, does not depend on the direction of deliberate, or endogenous, attention (Bertelson, Vroomen, de Gelder, & Driver, 2000). In the present study, a similar question concerning automatic, or exogenous, attention was examined. The experimental manipulation was based on the fact that exogenous visual attention can be attracted toward a singleton--that is, an item different on some dimension from all other items presented simultaneously. A display was used that consisted of a row of four bright squares with one square, in either the left- or the rightmost position, smaller than the others, serving as the singleton. In Experiment 1, subjects made dichotomous left-right judgments concerning sound bursts, whose successive locations were controlled by a psychophysical staircase procedure and which were presented in synchrony with a display with the singleton either left or right. Results showed that the apparent location of the sound was attracted not toward the singleton, but instead toward the big squares at the opposite end of the display. Experiment 2 was run to check that the singleton effectively attracted exogenous attention. The task was to discriminate target letters presented either on the singleton or on the opposite big square. Performance deteriorated when the target was on the big square opposite the singleton, in comparison with control trials with no singleton, thus showing that the singleton attracted attention away from the target location. In Experiment 3, localization and discrimination trials were mixed randomly so as to control for potential differences in subjects' strategies in the two preceding experiments. Results were as before, showing that the singleton attracted attention, whereas sound localization was shifted away from the singleton. Ventriloquism can thus be dissociated from exogenous visual attention and appears to reflect sensory interactions with little role for the direction of visual spatial attention. PMID- 11436736 TI - Segregation by color/luminance does not necessarily facilitate motion discrimination in the presence of motion distractors. AB - Under what circumstances is the common motion of a group of elements more easily perceived when the elements differ in color and/or luminance polarity from their surround? Croner and Albright (1997), using a conventional global motion paradigm, first showed that motion coherence thresholds fell when target and distractor elements were made different in color. However, in their paradigm, there was a cue in the static view of the stimulus as to which elements belonged to the target. Arguably, in order to determine whether the visual system automatically groups, or prefilters, the image into different color maps for motion processing, such static form cues should be eliminated. Using various arrangements of the global motion stimulus in which we eliminated all static form cues, we found that global motion thresholds were no better when target and distractors differed in color than when they were identical, except under certain circumstances in which subjects had prior knowledge of the specific target color. We conclude that, in the absence of either static form cues or the possibility of selective attention to the target color, features with similar colors/luminance polarities are not automatically grouped for global motion analysis. PMID- 11436737 TI - Efficient visual search by category: specifying the features that mark the difference between artifacts and animals in preattentive vision. AB - In this report, we explored the features that support visual search for broadly inclusive natural categories. We used a paradigm in which subjects searched for a randomly selected target from one category (e.g., one of 32 line drawings of artifacts or animals in displays ranging from three to nine items) among a mixed set of distractors from the other. We found that search was surprisingly fast. Target-present slopes for animal targets among artifacts ranged from 10.8 to 16.0 msec/item, and slopes for artifact targets ranged from 5.5 to 6.2 msec/item. Experiments 2-5 tested factors that affect both the speed of the search and the search asymmetry favoring detection of artifacts among animals. They converge on the conclusion that target-distractor differences in global contour shape (e.g., rectilinearity/curvilinearity) and visual typicality of parts and form facilitate search by category. We argue that existing theories are helpful in understanding these findings but that they need to be supplemented to account for the specific features that specify categories and to account for subjects' ability to quickly locate targets representing heterogeneous and formally complex categories. PMID- 11436738 TI - Feature-based control of attention: evidence for two forms of dimension weighting. AB - In three experiments, I examined whether prior knowledge of a target feature dimension is useful for guiding spatial attention to the target in a variety of tasks: visual search (Experiments 1A and 1B), texture segregation (Experiment 2), and visual enumeration (Experiment 3). Experiment 1A used a simple search task and found that reaction times for blocks in which a target was defined within a single feature dimension were shorter than those for blocks in which a target was defined across dimensions (within-dimension facilitation: WDF). Intertrial facilitation (ITF; Muller, Heller, & Ziegler, 1995), a dimension-based priming effect from one trial to the immediately following one, was also observed. Both WDF and ITF disappeared when the same stimuli were used under a compound search task (Experiment 1B), in which participants responded to an attribute of the target in a feature dimension different from its defining dimensions. Experiments 2 and 3 showed that WDF and ITF are not necessarily contingent upon each other: In a texture discrimination task, only WDF was found; in an enumeration task for six or seven targets, only ITF was found. These results show that the two forms of dimension weighting, WDF and ITF, are mediated by different mechanisms. WDF was eliminated when focal attention to targets was required, suggesting that feature-based modulation is limited as a source for controlling spatial attention (Kumada, 1999). ITF was correlated with type of response, suggesting dimension specific response mechanisms (Cohen & Shoup, 1997). PMID- 11436739 TI - Mental object rotation and the planning of hand movements. AB - Recently, we showed that the simultaneous execution of rotational hand movements interferes with mental object rotation, provided that the axes of rotation coincide in space. We hypothesized that mental object rotation and the programming of rotational hand movements share a common process presumably involved in action planning. Two experiments are reported here that show that the mere planning of a rotational hand movement is sufficient to cause interference with mental object rotation. Subjects had to plan different spatially directed hand movements that they were asked to execute only after they had solved a mental object rotation task. Experiment 1 showed that mental object rotation was slower if hand movements were planned in a direction opposite to the presumed mental rotation direction, but only if the axes of hand rotation and mental object rotation were parallel in space. Experiment 2 showed that this interference occurred independent of the preparatory hand movements observed in Experiment 1. Thus, it is the planning of hand movements and not their preparation or execution that interferes with mental object rotation. This finding underlines the idea that mental object rotation is an imagined (covert) action, rather than a pure visual-spatial imagery task, and that the interference between mental object rotation and rotational hand movements is an interference between goals of actions. PMID- 11436740 TI - The temporal cross-capture of audition and vision. AB - We report that when a flash and audible click occur in temporal proximity to each other, the perceived time of occurrence of both events is shifted in such a way as to draw them toward temporal convergence. In one experiment, observers judged when a flash occurred by reporting the clock position of a rotating marker. The flash was seen significantly earlier when it was preceded by an audible click and significantly later when it was followed by an audible click, relative to a condition in which the flash and click occurred simultaneously. In a second experiment, observers judged where the marker was when the click was heard. When a flash preceded or followed the click, similar but smaller capture effects were observed. These capture effects may reveal how temporal discrepancies in the input from different sensory modalities are reconciled and could provide a probe for examining the neural stages at which evoked responses correspond to the contents of conscious perception. PMID- 11436741 TI - Facilitative and inhibitory effects of cuing sound duration, intensity, and timbre. AB - Two experiments are reported in which the possibility that auditory attention may be controlled in a stimulus-driven manner by duration, intensity, and timbre cues was examined. In both experiments, listeners were presented with a cue followed, after a variable time period of a 150-, 450-, or 750-msec stimulus onset asynchrony (SOA), by a target. In three different conditions for each experiment, the duration, intensity, or timbre relation between the cue and the target was varied so that, on 50% of the trials, the two sounds were identical and, on 50% of the trials, the two sounds were different in the manipulated feature. The two experiments differed only in the judgment required, with listeners in Experiment 1 identifying the duration, intensity, or timbre of the target and listeners in Experiment 2 indicating whether the target incorporated a brief silent gap. In both experiments, performance was observed to depend on both the similarity of and the time between the cue and the target. Specifically, whereas at the 150 msec SOA performance was best when the target was identical to the preceding cue, at the 750-msec SOA performance was best when the cue and the target differed. This pattern establishes the existence of duration-, intensity-, and timbre-based auditory inhibition of return. The theoretical implications of these results are considered. PMID- 11436742 TI - Distinctiveness and serial position effects in tonal sequences. AB - The proportion-of-the-total-duration rule (Kidd & Watson, 1992) states that the detectability of a change in a component of a tonal sequence can be predicted by the proportional duration of the changed component relative to the length of the sequence as a whole. A similar viewpoint relies on temporal distinctiveness to account for primacy, recency, and other serial position effects in memory (Murdock, 1960; Neath, 1993a, 1993b). Such distinctiveness models predict that an item will be remembered if it is more distinctive along some dimension relative to possible competitors. Three experiments explored the relation between distinctiveness and proportional duration by examining the effects of the proportion of the total duration of a tone in a sequence, serial position, and interstimulus interval (ISI) on the detection of a change in one component of a tonal sequence. Experiment 1 replicated the basic effect with relatively untrained subjects and a fixed frequency difference. Experiment 2 showed that distinctiveness holds for tonal sequences and a same/different task. Experiment 3 combined the two to show that proportional duration, ISI, and position of the changed tone all contribute to discrimination performance. The present research combines theories that have been proposed in the psychophysics and memory fields and suggests that a comprehensive principle based on relative distinctiveness may be able to account for both perceptual and memory effects. PMID- 11436743 TI - Ability in perceiving nonnative contrasts: performance on natural and synthetic speech stimuli. AB - The perception of the distinction between /r/ and /l/ by native speakers of American English and of Japanese was studied using natural and synthetic speech. The American subjects were all nearly perfect at recognizing the natural speech sounds, whereas there was substantial variation among the Japanese subjects in their accuracy of recognizing /r/ and /l/ except in syllable-final position. A logit model, which additively combined the acoustic information conveyed by F1 transition duration and by F3-onset frequency, provided a good fit to the perception of synthetic /r/ and /l/ by the American subjects. There was substantial variation among the Japanese subjects in whether the F1 and F3 cues had a significant effect on their classifications of the synthetic speech. This variation was related to variation in accuracy of recognizing natural /r/ and /l/, such that greater use of both the F1 cue and the F3 cue in classifying the synthetic speech sounds was positively related to accuracy in recognizing the natural sounds. However, multiple regression showed that use of the F1 cue did not account for significant variance in natural speech performance beyond that accounted for by the F3 cue, indicating that the F3 cue is more important than the F1 cue for Japanese speakers learning English. The relation between performance on natural and synthetic speech also provides external validation of the logit model by showing that it predicts performance outside of the domain of data to which it was fit. PMID- 11436744 TI - The interface. PMID- 11436745 TI - Modified importance-performance analysis: an application to hospitals. AB - This paper seeks to investigate the reasons why, in an increasingly competitive health care milieu, patients choose certain hospitals over others. It introduces the modified importance-performance analysis technique and presents the method and findings of an empirical study which applied importance-performance analysis in a health care setting. The strategies derived from the study findings are discussed. PMID- 11436746 TI - Information as a distinct dimension for satisfaction assessment of outpatient psychiatric services. AB - The purpose of this study is to verify whether information on services would appear as a distinct dimension of satisfaction in a multidimensional scale. Data collection was performed in two phases: 263 patients received the original version of the questionnaire and 200 received an adapted version of the scale. The findings suggest that not only is it important to consider information as a distinct dimension of satisfaction but it is equally important to examine three categories, consisting of satisfaction with information on; patients' problems/illness; distinct treatment components such as medication and psychotherapy; and patients' treatment progress. PMID- 11436747 TI - The pursuit of excellence is not optional in the voluntary sector, it is essential. AB - This paper outlines the continuous improvement journey of a voluntary organisation. The significant level of organisational growth and improving quality of services described is clearly linked to the organisation's commitment to improvement. One of the approaches used in adopting a total quality culture was the EFQM model, specifically interpreted for the voluntary sector. Until recently such an approach would have been considered alien to the sector, not least because of its origins in the business community. This article contradicts this assumption. Issues addressed include how the improvement process is driven, the use of performance measurement, external verification and the difficulties in accessing sector appropriate benchmark data. It is suggested that the pursuit of excellence is no longer optional for the voluntary sector, it is essential. PMID- 11436748 TI - How service users become empowered in human service organizations: the empowerment model. AB - This article presents an empowerment model (EM) to be used by service users in human service organizations (HSOs). The EM is a structure for service user input to be integrated within the HSO at various administrative levels through a four step sequential process. The article fills a distinct void in the literature as there are numerous accounts about the importance of empowerment, but few on processes that need to be defined to operationalize the concept. Implications are directed toward administrators as they need to take leadership in implementing the EM in order to deliver more efficient and relevant services to their clients. PMID- 11436750 TI - Believing in and recognising healthcare excellence. PMID- 11436749 TI - Improving general practitioners' assessment and management of suicide risk. AB - Standards for assessing and managing suicide risk were developed and incorporated into a guidance manual for general practitioners. The effects of the manual on opinions and practice were evaluated using a quasi-experimental controlled before/after design, comparing participating general practitioners with others who did not use the manual. Thirty four general practitioners participated over a six-month period. The intervention group showed changes in perceptions, with increased satisfaction with their own methods and in their recognition and assessment of suicide risk. Their practice changed, with increased recording of relevant factors in notes. The comparison group did not change in these ways. It is concluded that general practitioners' practice and opinions in assessing and managing suicide risk were significantly improved using a minimal intervention. Given the importance of the topic and the small size of this study, further research is needed, examining changes in professional practice, knowledge and attitudes. PMID- 11436751 TI - "A little encouragement": health services and domestic violence. AB - The aim of this study was to identify those elements of health care which women who have experienced domestic violence consider to represent quality care and explore opportunities for an area health service to improve its service delivery to these women. To do this nine focus groups were conducted with six urban and three rural groups, recruited from Women's Support Services and Refuges, and 65 women participated. A trained facilitator conducted the focus groups with a representative from each participating organisation. Audiotapes were transcribed then coded using a thematic analysis approach. Analysis of the tapes revealed six opportunities for improvement of health services. Improve services to Aboriginal women; increase access to services; promote available services; improve the health service environment; educate service providers; and provide specialist health services. A major theme to emerge was the importance of a supportive environment where women were supported to make choices for their future. PMID- 11436752 TI - The nurse retention, quality of care and patient satisfaction chain. AB - This paper proposes an integrated approach to examining and dealing with the complex issue of nurse recruitment, retention, healthcare quality and patient satisfaction. The paper depicts and describes a generic conceptual framework or chain derived from a review of the literature on nurse recruitment and retention, service quality and human resource management. The chain is made up of the following components: NHS and Trust conditions and environment (internal quality) -service capability--nurse satisfaction--nurse retention--quality of patient care -patient satisfaction. The value of the chain is derived from its synthesis and display of the prime constituents or drivers of nurse satisfaction, quality of patient care and satisfaction. From this holistic picture it is possible for both national and local initiatives to be integrated in a mutually reinforcing way in order to achieve improvements in nurse recruitment, retention, quality of care and patient satisfaction. PMID- 11436753 TI - TQM in dental practice. AB - Society now expects more from its doctors and dentists, and these increasing demands can be summed up in one relatively new term for the medical profession: "quality management" (QM). Doctors and dentists formerly took the view that their performance could be assessed solely on the basis of their technical skills, ethics and expertise, but are now confronted with a new social imperative, from outside the profession--quality management. The author, prize-winner of the European Quality Award 2000 describes his approach to introduce the European Foundation for Quality Management (EFQM) Excellence Model in his dental practice. He shows that the EFQM model is well suited as a basis for a quality management system in healthcare. PMID- 11436754 TI - Professional integrity in the age of managed care: views of physicians. AB - At the end of 1997, one quarter of the American public used health maintenance organizations. This paper reports findings on physicians' perspectives on the role of managed care in their professional practices. The research data come from mailed surveys to physicians who are selected from the Cigna Directory of Physicians practicing in the State of Ohio. Subjects were asked to explain what managed care meant to them, and how long they have been practicing medicine. Questions also focused on professional autonomy, quality of care and career aspirations for the future. The results from the study suggest that managed care has had a negative impact on how physicians practice medicine. Several of our respondents reported that they are playing the role of a "double agent" and feel a sense of frustration in doing so. The degree of antipathy toward managed care differs between primary care physicians and specialists. PMID- 11436755 TI - Strategic alignment between senior and middle managers in local government and health. AB - The North West Change Centre at Manchester Business School has been developing collaborative programmes aimed at public sector middle managers. Evaluation recently completed, using follow-up interviews, studied the affect of the programmes on individuals, their organisation and their work intra organisationally. Reference is made to management theories on strategic alignment and cultural change. The results suggest that middle managers have been highly satisfied resulting in changed attitudes, greater confidence and worked-through project ideas being developed. However, a sense of being "out of alignment" with their organisation remains manifested in poor support for inter-agency working. A model of strategic alignment between middle and senior managers is demonstrated which illustrates the affects of strong and weak alignment and the implications for implementing modernised services. Recommendations are made on contextualising change, how managerial signalling can be improved, how pathways to changed organisational systems can be identified and how barriers to change can be overcome. PMID- 11436756 TI - Poorer half of the world can expect better health and prosperity within the next decade. PMID- 11436757 TI - Primary care trusts. Back to the future. AB - Primary care trusts, which went live last year, have had some success in attracting additional funding for new initiatives and tackling long-standing problems. There have been considerable delays in appointing executive committees. Management capacity is a huge problem. Many feel they do not have the staff to carry out basic administration. GPs find chairing PCTs very stressful. Major issues of development, roles and resources need to be addressed. PMID- 11436758 TI - Walk-in centres. Fast forward. AB - A walk-in centre opened in Liverpool in April 2000 is now seeing more than 3,000 patients a month. Many patients use the service for emergencies. More than half those attending do not need referral elsewhere. Initial wariness on the part of local GPs to the project seems to have been largely overcome. The centre opens at 7 am but there is often a peak in demand at 9 am. PMID- 11436759 TI - Medical secretaries. Making the grade. AB - A review of the role of medical secretaries in a surgical directorate has reduced turnover among this group to zero. Previously the directorate had difficulty with recruitment and often had to rely on agency staff. The review led to the introduction of the new post of business coordinator, and all medical secretaries in the directorate have now achieved this level. The review led to an expanded role for medical secretaries which has increased their job satisfaction. PMID- 11436760 TI - My brilliant career--finance. Adding up from the start. Interview by Seamus Ward. PMID- 11436761 TI - Data briefing. Inverse care law. PMID- 11436762 TI - Compensation. The blame drain. PMID- 11436763 TI - Income generation. Brain powers. PMID- 11436764 TI - Product liability. Fault lines. PMID- 11436765 TI - Reducing medical errors. Flying start. PMID- 11436766 TI - Corporate killing. Heated debate. PMID- 11436767 TI - Climbing the ladder: a step by step approach to international guidelines for water recycling. AB - Given the pressures on the world's freshwater resources, recycled water is a valuable resource. Recycled water can increase the reliability of water supply because it is an independent source of water. Water recycling requires effective measures to protect public health and the environment. In the absence of comprehensive international guidelines, different countries have developed different approaches to managing water recycling depending on the understanding of the health risks, their individual economic circumstances, and affordability. Approaches vary between high technology/high cost/low risk and low technology/low cost/controlled risk. Furthermore, differences occur between countries and within individual countries. Inconsistencies can often be traced to lack of a unified scientific position on health effects. These inconsistencies increase public concerns about health risks and may give rise to conservative controls on responses to water recycling projects that some countries may be unable to afford. In this paper, an international panel of authors discusses how the different water recycling approaches might be linked together into international water recycling guidelines. These guidelines would incorporate a uniform approach to assessing hazards and risks while providing flexibility for individual countries to vary requirements to suit local circumstances of affordability and risk. The authors propose a framework of guidelines in which individual countries can progressively improve recycled water quality as lower risk levels become more affordable. The authors argue that a uniform international approach will result in a number of benefits including a better focus on risk management, better targeted research and development efforts and greater public confidence in water recycling. The authors invite discussion on the concepts put forward in the paper. PMID- 11436768 TI - From sewage water treatment to wastewater reuse. One century of Paris sewage farms history. AB - The irrigation fields of Paris have been used for 100 years. Their soils mainly contain heavy metals in the topmost layer. Metals come from raw sewage as well as from digested sludge of biological treatment plants which have been diluted for years in raw water. Vegetables that are cultivated in the irrigation fields concentrate metals but their average contents, however, are lower than the recommended limit values. Some vegetables concentrate more specifically one type of metal. Corn seeds accumulate less metal than green vegetables. The SIAAP keeps operating irrigation fields by delivering clariflocculated water with a low metal content from the new Seine Centre plant, with the purpose of keeping some 2,000 ha of green zone in an otherwise heavily constructed area and to prevent a metal release from the soil should irrigation be interrupted. Maintaining irrigation fields also relieves the biological treatment plant and then contributes to preserve the quality of the Seine river, especially in summer. PMID- 11436769 TI - Cultural eutrophication control through water reuse. AB - The increasing use of mineral fertilisers over the last decades has contributed to the appearance of numerous cases of water eutrophication, a new form of water pollution. The starting point of eutrophication is the increase of nutrient concentration (nitrogen and phosphorus) in a water mass, which is subsequently followed by an uncontrolled growth of primary producers and episodes of oxygen depletion due to microbial decomposition of algal organic matter. The excess nutrient loads reaching surface waters are usually associated to discharges from anthropogenic activities, which normally involve direct water usage instead of reuse of reclaimed effluents. Agriculture activities and livestock breeding are two of the main nutrient sources responsible for water eutrophication, as well as human--urban and industrial--wastewater discharges. Wastewater reclamation and reuse can be a suitable strategy for preserving the quality of natural waters, by suppressing effluent discharges and the associated nutrient contributions to receiving waters. Reuse of reclaimed water for agricultural and landscape irrigation as well as for environmental enhancement offers an adequate strategy for preserving natural water systems from eutrophication. PMID- 11436770 TI - Reuse of reclaimed wastewater for golf course irrigation in Tunisia. AB - In Tunisia, golf courses are irrigated with secondary treated effluent stored in landscape impoundments. The impact of the conveyance and storage steps on the physical-chemical and biological quality of irrigation water was evaluated on three golf courses over two years. It was found that the water quality varies all along the water route, from the wastewater treatment plant up to the irrigation site: nutrient and bacteria contents decreased along the route in the three cases. This variation depends on the wastewater quality, the length of the pipes conveying water, the number of regulation reservoirs and ponds, the water residence time in pipes, reservoirs and ponds, and the operation of the ponds. The bacteriological quality of irrigation water deteriorates during the irrigation period in the three golf courses as the ponds are operated as continuous flow reactors. The results obtained in this study indicate the inability of golf water supplies, as currently managed, to properly sanitize reclaimed wastewater and meet target quality criteria recommended by WHO (1989) for water intended for recreational use. For a safe reuse of reclaimed wastewater for golf course irrigation, changes in the design and operation of the ponds should be planned or additional treatment steps provided. PMID- 11436771 TI - Ammonia removal in a deep reclaimed wastewater reservoir before agricultural reuse. AB - An experimental study has been carried out to determine the ammonia removal in a deep reclaimed wastewater reservoir of 50,000 m3. The reservoir is part of the Reuse System of Reclaimed Wastewater of South Tenerife (Spain). The study has been conducted under batch mode operation, in three different periods (winter, spring-summer and winter-spring), with an average storage time of 85 days. Vertical profiles of temperature, dissolved oxygen and pH were determined together with NH3-N, NO2-N and NO3-N concentrations at different points and different depths in the reservoir. Maximum removal efficiency was obtained during the winter periods, in absence of stratification and with good mixing conditions throughout the water column. During these periods, nitrification took place in some extension, favoring the ammonia removal. Average NH3-N concentrations in the reservoir were calculated and apparent first-order rate constants were determined for different stratification conditions. Although ammonia nitrogen could be used as nutrient in the agricultural reuse, its removal from reclaimed wastewater could be useful in order to diminish the chlorine needs for disinfection. PMID- 11436772 TI - How partial nitrification could improve reclaimed wastewater transport in long pipes. AB - Reclaimed wastewater transport is studied in a concrete-lined cast iron pipe, where a nitrification-denitrification process occurs. The pipe is part of the Reuse System of Reclaimed Wastewater of South Tenerife (Spain), 0.6 m in diameter and 61 km long. In order to improve wastewater quality, at 10 km from the inlet there is injection of fresh water saturated in dissolved oxygen (DO), after which a fast nitrification process usually appears (less than two hours of space time). The amount of oxidized nitrogen compounds produced varies between 0.8 and 4.4 mg/l NOx(-)-N. When DO has disappeared, a denitrification process begins. The removal of nitrite is complete at the end of the pipe, whereas the nitrate does not disappear completely, leaving a concentration of about 0.4-0.5 mg/l. For a COD/NOx(-)-N ratio higher than 5, a first order nitrification rate in NOx(-)-N has resulted, with the constant k20 = 0.079 h-1, for a NOx(-)-N concentration range of 0.8-4.4 mg/l. Finally the following temperature dependency for the first order denitrification rate constant has been found: k = k20 x 1 x 15T-20. Although nitrogen could be used as nutrient in the agricultural reuse, its removal from reclaimed wastewater could be useful in order to diminish the chlorine needs for reclaimed wastewater disinfection. PMID- 11436773 TI - Secondary wastewater disposal for crop irrigation with minimal risks. AB - A critical objective for any wastewater reuse program is to close the gap between supply of and demand for water and to minimize health and environmental hazards. Thus, the effects of treated effluent on crops, soils and community health must be considered carefully. When applying wastewater to soil-plant systems, it is to be noted that the passage of water through the soil reduces considerably the number of microorganisms carried out by the reclaimed wastewater. Nevertheless, there is a need to study the real rate of organism decay subject to water quality, soil and vegetable characteristics, and irrigation method. The aim of this work is to determine the fate of the fecal coliforms, coliphages F+ and CN13, and helminth eggs survival during the application of reclaimed wastewater in a vineyard orchard near the City of Arad (Israel) via onsurface and subsurface drip irrigation systems. Wastewater obtained from a stabilization pond, and soil samples were tested and an important decrease of microorganisms was reached in both cases, with the better values obtained with the sub-surface drip irrigation system. PMID- 11436774 TI - Wastewater quality relationships with reuse options. AB - The trend towards reuse of effluent for land application of domestic and industrial wastewater is driven by the need to maximise limited water resources and benefit from the plant nutrients available in the effluent. Of significant impact upon the value of the wastewater for reuse is its chemical properties as well as biochemical oxygen demand and suspended solids. While the sewage treatment plant is expected to treat all wastewater received to a minimum environmental standard, no efforts are given to reducing the chemical load derived from uses of chemicals in the domestic household. That the regulation of industry and commerce far exceeds those of the combined effects of thousands of household is remiss of environmental regulators. This paper examines the results of research into the more common sources of chemical additives to the wastewater stream. Twenty five potable water supplies are examined for their salt load, 20 liquid and 40 powder laundry detergents and five dishwashing products were used to simulate discharges to the sewer, measured for their phosphorus, salt and sodium concentrations. The results of the research indicate that choices in the products available for general use within the house can be made only where product labelling and consumer education is improved. Technical improvements in wastewater treatment are not the answer. The improvement in effluent quality will have significant beneficial effects upon land application areas and expand the range of reuse options available for commercial operations. PMID- 11436775 TI - Removal of microorganisms in different stages of wastewater treatment for Mexico City. AB - In this study the removal of a diversity of microorganisms during different stages of treatment was evaluated. The process scheme consisted of Advanced Primary Treatment (APT), sand filtration, and chlorine disinfection. During the first 2 stages, fecal coliforms were reduced from 1.24 x 10(9) to 1.2 x 10(8) MPN/100 mL and helminth ova were reduced to less than 1 ova/L. Nevertheless, to obtain an effluent water quality that meets the microbiological standards recommended by the World Health Organization (1989) and the Mexican norm for water destined for agricultural reuse it was necessary to disinfect the effluent with a chlorine dose of 12 mg/L for a 3 hour contact time. Under these conditions, fecal coliforms were reduced from 1.2 x 10(9) to 5.8 x 10(1) MPN/100 mL, Salmonella spp. from 5.0 x 10(8) MPN/100 mL to below the detection limit, Pseudomonas aeruginosa from 2.0 x 10(5) MPN/100 mL to below the detection limit, and protozoan cysts from 1052 to 31 cysts/L. Regrowth of bacteria was never observed during the 48 hours period following disinfection. The active chlorine was primarily in the form of chloramines thus reducing the formation of other potentially carcinogenic disinfection byproducts. PMID- 11436776 TI - Wastewater disinfection by low-pressure UV and ozone: a design approach based on water quality. AB - Disinfection processes are known to be very sensitive to wastewater quality. This paper discusses the parameters that impact the UV light (UV) and ozone disinfection processes and the related mechanisms based on literature review. Low pressure UV and ozone technologies were investigated on effluents that covered a wide range of water quality. The results are given in terms of design doses required to meet three major disinfection standards. Both processes were found eligible for the majority of effluents tested. Although cost-effectiveness is usually considered more favourable to UV, the ozone alternative should be examined in cases such as the disinfection of low-quality effluents or large treatment plants. Ozonation was also found capable of meeting the stringent Title 22 standard with no coagulation at a dose of 10 mg/l. PMID- 11436777 TI - Water reuse in Japan. AB - Even though Japan has mean annual precipitation of 1,714 mm and hundreds of dams and reservoirs constructed, frequent and severe droughts have occurred in wide regions of the country. Because of rapid economic growth and concentrations of population in urban areas, water demands in large cities have stressed reliability of water supply systems and necessitated the development of new water resources with considerable economic and environmental costs. To alleviate these situations, wastewater reclamation and reuse have been implemented widely in major cities. This paper summarizes the current status of water reuse in Japan and discusses dominant uses of reclaimed water, emphasizing non-potable urban applications such as toilet flushing, industrial reuse, and environmental water. PMID- 11436778 TI - UV disinfection for reuse applications in North America. AB - In an effort to conserve and protect limited water resources, the States of Florida and California have actively promoted wastewater reclamation and have implemented comprehensive regulations covering a range of reuse applications. Florida has a semi-tropical climate with heavy summer rains that are lost due to run off and evaporation. Much of California is arid and suffers periodic droughts, low annual rainfall and depleted ground water supplies. The high population density combined with heavy irrigation demands has depleted ground water supplies resulting in salt-water intrusion. During the past decade, Florida reuse sites have increased dramatically from 118 to 444 plants representing a total flow capacity of 826 MGD. California presently has over 250 plants producing 1 BGD with a projected increase of 160 sites over the next 20 years. To prevent the transmission of waterborne diseases, disinfection of reclaimed water is controlled by stringent regulations. Many states regulate wastewater treatment processes, nutrient removal, final effluent quality and disinfection criteria based upon the specific reuse application. As a rule, the resulting effluents have low turbidity and suspended solids. For such effluents, UV technology can economically achieve the most stringent disinfection targets that are required by the States of California and Florida for restricted and unrestricted reuse. This paper compares UV disinfection for wastewater reuse sites in California and Florida and discusses the effect of effluent quality on UV disinfection. PMID- 11436779 TI - Bacterial responses to ultraviolet irradiation. AB - The UV dose-response behavior of laboratory cultures of waterborne bacteria were examined for UV doses ranging from ca. 0-100 mW.s/cm2 using a collimated-beam reactor. Specific physiological responses measured in these tests included viability (ability to reproduce) and respiration (oxygen uptake rate). The results of these exposures indicated that resistance to UV-imposed loss of viability in E. coli cultures can be partially attributed to agglomeration during the irradiation process. From these results, it is conjectured that a bacterial population may be comprised of two sub-populations: one with low resistance (discrete or paired cells) and a second with high resistance (bacterial aggregates). A small fraction of the high-resistance portion of the population appears to be essentially unaffected by UV irradiation, thereby causing a discontinuity in the measured dose-response behavior. Moreover, the dose-response behavior of the highly resistant fraction is variable and difficult to describe quantitatively. The basis of these statements and most information in the literature is microbial viability as quantified by the membrane filtration assay. In contrast to these findings, the results of analyses for bacterial activity (respiration) suggest that comparatively little change in the population can be found to result from UV irradiation. This suggests that UV radiation accomplishes inactivation of the bacteria, but does not "kill" the bacterial cells per se, thereby highlighting the importance of considering bacterial repair processes in the design of UV disinfection systems. PMID- 11436780 TI - Faecal bacteria and bacteriophage inactivation in a full-scale UV disinfection system used for wastewater reclamation. AB - A study was carried out to compare the inactivation of faecal bacteria and one type of bacteriophage in a full-scale UV disinfection system. The system is part of a water reclamation facility for effluent reuse in golf course and agricultural irrigation. Influent and effluent samples were taken over two sampling periods (three consecutive days in July and one day in August), with three different UV doses applied each day (ranging from 10 to 40 mW.s/cm2 and 20 to 80 mW.s/cm2 in July and August, respectively). Effluent samples were also taken from a chlorine disinfection channel (5 mg Cl2/L dose) operating in parallel to the UV system. Total coliforms (TC), faecal coliforms (FC), faecal streptoccoci (FS) and somatic coliphages (SC) were measured in each sample. F specific RNA bacteriophages and bacteriophages of Bacteroides fragilis were also measured one day in July. The decay ratio observed for all the microorganisms was similar when UV doses applied were low (July), ranging from 1.15 to 1.25 log units. This suggests that bacterial indicators may be suitable for virus inactivation control when low UV doses are applied; however, such low doses are inadequate to achieve effluent quality requirements for unrestricted irrigation. At higher UV doses (August), decay ratios for TC and FC were 3.1 and 2.8 log units respectively, indicating that they were more susceptible to UV exposure than SC and FS, with decay ratios of 2.6 and 1.0 log-units, respectively. Nevertheless, these higher doses were also inadequate to achieve water quality requirements for unrestricted irrigation. The decay ratio of SC during chlorine disinfection was clearly lower than that of the other microorganisms. Bacteriophages of Bacteroides fragilis were more resistant to UV disinfection than SC and F-specific RNA. In fact, bacteriophages of Bacteroides fragilis were not affected during UV exposure. A UV dose ranging from 40 to 80 mW.s/cm2 marks the borderline beyond which inactivation rates of SC are clearly lower than those of bacterial indicators. PMID- 11436781 TI - Biological control tools for wastewater reclamation and reuse. A critical review. AB - The health related risk of wastewater reclamation and reuse is usually defined by laws, rules or regulations by using only biological tools; i.e. bacteria, viruses, or other pathogens or indicators determination. Those determinations exert some influence in the costs of the mentioned practices, and it seems probable that in the near future more determinations will be required. Nevertheless, a total indication of wastewater biological quality is not given by such organisms; in fact, long-term toxicity is not detected by such means. The future of biological control of reclamation and reuse systems and the price associated to such determinations is examined through a practical application case. PMID- 11436782 TI - Feasibility of the membrane bioreactor process for water reclamation. AB - The feasibility of the membrane bioreactor (MBR) process for water reclamation was studied. Process evaluation was based on the following: literature review of MBRs, worldwide survey of MBRs, and preliminary costs estimates. The literature review and the survey have shown that the MBR process offers several benefits over the conventional activated sludge process, including: smaller space and reactor requirements, better effluent water quality, disinfection, increased volumetric loading, and less sludge production. The MBR process can exist in two different configurations, one with the low-pressure membrane modules replacing the clarifier downstream the bioreactor (in series), and the second with the membranes submerged within the bioreactor. Four major companies are currently marketing MBRs while many other companies are also in the process of developing new MBRs. The MBR process operates in a considerably different range of parameters than the conventional activated sludge process. The preliminary cost evaluation has shown that the MBR process is cost competitive with other conventional wastewater treatment processes. PMID- 11436783 TI - Advanced biological unit processes for domestic water recycling. AB - The potential of advanced biological unit operations for the recycling of grey and black waters has been evaluated. The membrane bioreactor (MBR) demonstrated the greatest efficacy towards water recycling in terms of all the quality determinants. Both the biologically aerated filter (BAF) and the MBR were able to effectively treat the organic and physical pollutants in all the types of wastewater tested. The main difference was observed in terms of the microbiological quality, measured as total coliforms. The open bed structure of the BAF enabled passage of coliforms whereas the complete barrier of the MBR produced a non detectable level in the effluent. The MBR process complied with commonly adopted water recycling quality standards for the all determinants during the grey water trials and failed only in terms of total coliform counts once black water had been introduced into the feed. The MBR was seen as a particularly suitable advanced biological process as it was very effective at stabilising out the considerable load variations encountered during the trial. PMID- 11436784 TI - Evaluation of an integrated membrane system for water repurification. AB - An integrated membrane system was evaluated for water repurification. Performance evaluation of the membrane system was based on three criteria: flux and fouling, disinfection capability, and rejection of pollutants. Minimal membrane fouling was observed for all of the membranes employed in the study. Significant contaminant rejection was also achieved by the membrane system purifying the reclaimed water to meet and exceed drinking water standards. Wide range of virus rejection was observed for the membranes, which was dependent on the membrane type, manufacturer, and the fouling status. Overall, the results of this study demonstrated that the integrated membrane system is a very effective and reliable process for water repurification. PMID- 11436785 TI - Membrane filtration of wastewater effluents for reuse: effluent organic matter rejection and fouling. AB - The reuse of treated wastewater to augment natural drinking water supplies is receiving serious consideration. Treatment of secondary and tertiary effluent by membrane filtration was investigated by assessing nanofiltration (NF) membrane and ultrafiltration (UF) membranes in bench-scale experiments. It was found that secondary and tertiary effluent contained high concentration of effluent organic matter (EfOM), contributing EfOM-related fouling. Flux decline and EfOM rejection tests were evaluated, using a dead-end stirred cell filtration unit. Surface charge and molecular weight cut-off (MWCO) of membranes were significant factors in membrane performance including permeability and EfOM-rejection. PMID- 11436786 TI - Pilot scale nanofiltration membrane separation for waste management in textile industry. AB - This paper presents the pilot scale membrane separation studies on dyehouse effluents of textile industry. Nanofiltration (NF) membranes which have 2 m2 of surface area were evaluated for membrane fouling on permeate flux and their suitability in separating COD, color and conductivity in relation to operating pressure and feed concentration from textile industry dyehouse effluents. Successive batch runs demonstrated that any serious membrane fouling was not experienced for NF membrane tested in treating this type of wastewater. The permeate flux was found to increase significantly with operating pressure. Flux decreased with increasing recovery rate. The overall removal efficiencies of COD, color and conductivity were found as greater than 97%. COD was lower than 10 mg/l at 12 bar pressures. Permeate COD was also increased with increasing recovery and COD was 30 mg/l with recovery of 80%. Almost complete color removal was achieved with nanofiltration membrane. Color value was also decreased from 500 Pt-Co to 10 Pt-Co unit. This significant reduction in color and COD makes possible the recycle of the permeate in the dyehouse. Permeate conductivity was decreasing with increasing pressure and retention of conductivity increases with increasing pressures. This phenomenon is expected from the analysis of conductivity mass transport model. Economical analysis have been done and the total estimated cost will be 0.81$/m3 based on 1000 m3/day of and this value is very economical for Istanbul City due to increasing industrial water supply tariffs. PMID- 11436787 TI - New parameter for monitoring fouling during ultrafiltration of WWTP effluent. AB - Variations in water quality of waste water treatment plant (WWTP) effluent complicate ultrafiltration of this feed water. Traditional parameters do not provide sufficient information to explain the fouling of membranes during ultrafiltration of WWTP effluent. New parameters for measuring and monitoring the fouling potential of feed water for ultrafiltration membranes need to be developed. The normalised membrane fouling index for ultrafiltration membranes (MFI-UFn) can be used as such and is according to the cake filtration theory calculated from the ratio of filtration time and filtration volume as a function of the filtration volume. MFI-UFn can be calculated from both experiments with constant Trans Membrane Pressure (TMP) and from experiments with constant flux. This parameter can also be calculated independent of the scale of the experiment. Results show that differences in fouling potential can be measured for various feed waters using the same membrane type and for various membrane types using the same feed water. Variation in feed water quality leads to a deviation of the MFI UFn, as was found especially for WWTP effluent. The applied TMP influences the value of the MFI-UFn, indicating cake compression when applying a higher TMP. MFI UFn can be used to identify the effect of pre-treatment methods, which is useful when using WWTP effluent as feed water for an ultrafiltration processes. PMID- 11436788 TI - Water quality assessment for indirect potable reuse: a new methodology for controlling trace organic compounds at the West Basin Water Recycling Plant (California, USA). AB - The West Basin Water Recycling plant (California; USA) was built to increase the region's water resource availability. The plant influent is produced at Los Angeles Hyperion wastewater treatment plant and is treated through two parallel treatment processes depending on the end use: (1) Title 22 water for industrial and urban use, and (2) barrier treatment for groundwater recharge. A new methodology was applied to monitor the fate of base neutral compounds in the water barrier treatment train. The methodology included large sample volumes coupled with integrated chromatographic analysis (ICA). Data indicated a 25% increase in concentration of base neutral compounds after RO pretreatment, followed by a 70% removal efficiency after RO. The increase in concentration after RO pretreatment appears to be linked to the use of lime clarification. PMID- 11436789 TI - Role of water reuse for enhancing integrated water management in Europe and Mediterranean countries. AB - Recycling water is an important aspect of water resource and environment management policies, ensuring reliable alternative water resources, reducing environmental pollution and achieving a more sustainable form of development. This paper focuses on wastewater reuse as a strategy for integrated water management. Key economic, financial, regulatory, social and technical factors that help to make water reuse projects successful are reviewed. Selected examples from Northern and Western Europe and arid and semi-arid Mediterranean regions illustrate the contribution of wastewater reuse to integrated management of water resources. PMID- 11436790 TI - Demonstration, the solution to successful community acceptance of water recycling. AB - The Department of Natural Resources in Queensland, Australia are presently carrying out a comprehensive Strategy called the Queensland Water Recycling Strategy (QWRS) to determine future Government directions in the whole area of water recycling. This strategy is considering the beneficial use of all waste streams such as domestic sewage, industrial and agricultural wastes, as well as urban stormwater. Following a workshop held during the initial phase of the strategy it was determined that a high priority must be given to the demonstration of recycling practices not being utilised in the State, or presently being practiced in an unsustainable manner. Three separate types of recycling projects are being carried out, the first being based on demonstrating recycling on a large new urban development close to Brisbane, the second associated with demonstrating the complex treatment processes associated with the higher levels of recycling, and the third associated with demonstrating community based recycling schemes. PMID- 11436791 TI - Source water impact model (SWIM)--a watershed guided approach as a new planing tool for indirect potable water reuse. AB - The scope of this study was to develop a model to assess the impact of source water quality on reclaimed water used for indirect potable reuse. The source water impact model (SWIM) considered source water qualities, water supply distribution data, water use and the impact of wastewater treatment to calculate reclaimed water quality. It was applied for sulfate, chloride, and dissolved organic carbon (DOC) at four water reuse sites in Arizona and California. SWIM was able to differentiate between the amount of salts derived by drinking water sources and the amount added by consumers. At all sites, the magnitude of organic residuals in reclaimed water was strongly effected by the concentration of organics in corresponding water sources and effluent-derived organic matter. SWIM can be used as a tool to predict reclaimed water quality in existing or planned water reuse systems. PMID- 11436792 TI - Reuse of heat energy in wastewater: implementation examples in Japan. AB - Sewage and treated water can be a heat source in urban area due to large heat capacity, thus recovery and reuse of its energy is one of the most desirable plans for the sewerage system. In this paper, characteristics of heat energy in wastewater, reuse plans, and some experiences in Japan are presented. Full-scale reuse projects for heating and cooling in the Tokyo Metropolitan Districts and project for melting snow in Sapporo City are discussed. The key factors found in experience of Tokyo were setting the heat pumps near the demand points and the technical developments of equipment to prevent system from clogging, corrosion, and decrease in the heat transfer efficiency. It was also found through the project for melting snow in Sapporo that the key factor in public acceptance was the multi-purpose use of the sewerage system both for melting snow in winter and retaining rain water in summer. PMID- 11436793 TI - Water recycling at the Millennium Dome. AB - Thames Water is working with the New Millennium Experience Company to provide a water recycling system for the Millennium Dome which will supply 500 m3/d of reclaimed water for WC and urinal flushing. The system will treat water from three sources: rainwater--from the Dome roof greywater--from handbasins in the toilet blocks groundwater--from beneath the Dome site The treatment technologies will range from "natural" reedbeds for the rainwater, to more sophisticated options, including biological aerated filters and membranes for the greywater and groundwater. Pilot scale trials were used to design the optimum configuration. In addition to the recycling system, water efficient devices will be installed in three of the core toilet blocks as part of a programme of research into the effectiveness of conservation measures. Data on water usage and customer behaviour will be collected via a comprehensive metering system. Information from the Dome project on the economics and efficiency of on-site recycling at large scale and data on water efficient devices, customer perception and behaviour will be of great value to the water industry. For Thames Water, the project provides vital input to the development of future water resource strategies. PMID- 11436794 TI - Chemical oxidation of organic matter in secondary-treated municipal wastewater by using methods involving ozone, ultraviolet radiation and TiO2 catalyst. AB - The efficiencies of chemical oxidation of organic matter in genuine secondary treated municipal wastewater by using O3, O3/UV, O3/TiO2, UV/TiO2 and O3/UV/TiO2 methods were compared experimentally. Effects of carbonates on these efficiencies were also investigated. Elimination of carbonates from the water increased TOC reduction efficiency of all the methods. The effect of the carbonates on the O3 method was less significant than that on the other methods. The O3/UV method was the most effective for reducing TOC under both carbonate-present and decarbonated conditions. The catalytic ozone process (O3/TiO2) showed a tendency to increase the ozonation efficiency at the beginning of the reaction under the decarbonated condition, but the addition of the TiO2 catalyst inhibited the ozonation under the carbonate-present condition. The photocatalytic oxidation process (UV/TiO2) did not reduce TOC as much as the other methods, but it could reduce TOC over a long time. And the O3/UV/TiO2 method did not show a synergistic effect of ozonation and photocatalytic oxidation. Furthermore, zonation prior to coagulation increased TOC reduction efficiency of coagulation, but this effect was also affected by the carbonates in water. PMID- 11436795 TI - Particle size analysis as a tool for performance measurements in high rate effluent filtration. AB - In the Netherlands almost all wastewater treatment plants have been redesigned and adapted in order to remove nitrogen, phosphorus and suspended solids to a very low level. The improved effluent quality leads to a growing interest in the reuse of effluent of the modernised wastewater treatment plants. This again results in investigations on filtration techniques as deep bed filtration and membrane filtration. At the wastewater treatment plant Ede research was done on deep bed filtration in order to develop relations between particle removal and filter performance and to explore ways of optimization. The results of the experiments are rather typical for effluent of modern Dutch wastewater treatment plants. The very low concentrations of suspended solids and precipitable substances result in poor flocculating properties. From turbidity measurements it may be concluded that the best results were obtained with a dosage of flocculant. However, the particle size measurements indicated the opposite. Suspended solids calculations, based on the particle volume distributions, showed a better removal without a dosage of flocculant. From this it is concluded that a dosage of coagulant (Fe3+ or Al3+) has an adverse effect on the removal efficiencies even at low dosages (1 mg/l). PMID- 11436796 TI - Improved O3/H2O2 oxidation process for wastewater reclamation. AB - A continuous two step O3/H2O2 oxidation system, in which the waste gas from the former reactor was reused in the latter step with an ejector, was established for the purpose of wastewater reclamation in electronics industries. ORP monitor was combined into the system to obtain the optimum ratio of H2O2 to O3 by automatically adjusting the dosage of H2O2, and ion exchange resin was used for removing organic acids formed during oxidation. The effectiveness of the O3/H2O2 oxidation system for TOC removal was investigated under various conditions by using IPA, DMSO, and NMP as model substances. The optimum H2O2/O3 for oxidizing IPA was in a range above 0.15, and it increased with the increase of O3 dose. A sudden drop of ORP from ca. 800 mV to a little more than 200 mV was observed when H2O2/O3 was approaching the optimum point from the lower end. An ORP between 250 mV and 300 mV or 800 mV and 900 mV was proper for H2O2/O3 control. The treatability of the model substances was in an order of IPA << DMSO < NMP, being consistent with the magnitude of the energy of the highest occupied molecular orbital (epsilon HOMO). The two step treatment system developed in this study was found to be more efficient than the conventional single step system. With the help of H2O2 dosing control and two step treatment using waste O3, the system could save about 40% O3 in comparison with the conventional O3/H2O2 oxidation process. PMID- 11436797 TI - Removal of color substances using photocatalytic oxidation for membrane filtration processes. AB - This study aims to remove the color substances, which normally cause difficulties in membrane filtration processes due to fouling using heterogeneous UV/TiO2/H2O2 reactor. It is confirmed that the technique used in this study was effective to remove TOC at 38% and color400 at 89% within 150-min irradiation. The experiment results showed that low concentrations of hydrogen peroxide dosage (less than 0.016 M) to UV/TiO2 system accelerated the TOC and Color400 removal rate from 9% to 38% and 40% to 89% respectively, while over-dosage made this positive effect decline. The humic acid solution treated by photo catalytic oxidation in UV/TiO2/H2O2 reactor did not change the zeta potential on membrane surface and membrane rejection rate. PMID- 11436798 TI - The effects of water reclamation technologies on biological stability of industrial water. AB - A modified Assimilable Organic Carbon (AOC) procedure was adopted in conjunction with Heterotrophic Plate Count (HPC) method to assess the effect of Single Effect Distillation (SED) and Reverse Osmosis (RO) lab-scale systems on the biological stability of industrial water. Industrial water was collected from a local Industrial Water Works, pre-treated with alum coagulation and cartridge filtration, before being subjected to advanced water treatment. The results obtained in this study indicated that AOCs in the SED product water were in the range of 70-80 micrograms acetate-C/L, while those in the RO product water ranged from 30-40 micrograms acetate-C/L in the 15-min permeate to 55-65 micrograms acetate-C/L in the 3-hr permeate. The above findings suggested that product water of both systems were potentially biologically unstable and would likely lead to bacteria regrowth during its distribution and storage. Removal efficiencies of lab-scale RO and SED systems on AOC were as high as 90%, dependent on the concentration of AOC-NOX in the industrial water. The RO system had much higher organic removal efficiencies in terms of AOC and DOC than the SED system. Organics removed from both feed waters were found to be concentrated in the brine water and rejected water in SED and RO systems respectively. PMID- 11436799 TI - The filter system for tertiary treatment of sewage effluent by land application- its performance in a subtropical environment. AB - FILTER is an innovative, CSIRO developed system for treating effluent using high rate land application and subsequent effluent recapture via a closely spaced, subsurface drainage network. We report on the summer performance of a FILTER system established in a subtropical environment on a relatively impermeable swelling clay soil underlain by a deep regional water table. Using secondary treated sewage effluent, the FILTER system produced effluent of tertiary nutrient standards (< or = 5 mg/L TN; < or = 1 mg/L TP), with salinity levels suitable for subsequent irrigation reuse (EC < or = 2.5 dS/m). Removal of faecal coliforms was considerably less effective. The hydraulic loading rate achieved was about two and a half times large than conventional irrigation demand, but this was associated with high deep percolation losses (c 3 mm/day). Comparisons are made with the original FILTER system developed and tested by Jayawardane et al. in temperate Australia. Suggestions are made for modifications to, and further testing of FILTER in a subtropical environment. PMID- 11436800 TI - Water quality transformations during soil aquifer treatment at the Mesa Northwest Water Reclamation Plant, USA. AB - Water quality transformations during soil aquifer treatment at the Mesa Northwest Water Reclamation Plant (NWWRP) were evaluated by sampling a network of groundwater monitoring wells located within the reclaimed water plume. The Mesa Northwest Water Reclamation Plant has used soil aquifer treatment (SAT) since it began operation in 1990 and the recovery of reclaimed water from the impacted groundwater has been minimal. Groundwater samples obtained represent travel times from several days to greater than five years. Samples were analyzed for a wide range of organic and inorganic constituents. Sulfate was used as a tracer to estimate travel times and define reclaimed water plume movement. Dissolved organic carbon concentrations were reduced to approximately 1 mg/L after 12 to 24 months of soil aquifer treatment with an applied DOC concentration from the NWWRP of 5 to 7 mg/L. The specific ultraviolet absorbance (SUVA) increased during initial soil aquifer treatment on a time-scale of days and then decreased as longer term soil aquifer treatment removed UV absorbing compounds. The trihalomethane formation potential (THMFP) was a function of the dissolved organic carbon concentration and ranged from 50 to 65 micrograms THMFP/mg DOC. Analysis of trace organics revealed that the majority of trace organics were removed as DOC was removed with the exception of organic iodine. The majority of nitrogen was applied as nitrate-nitrogen and the reclaimed water plume had lower nitrate-nitrogen concentrations as compared to the background groundwater. The average dissolved organic carbon concentrations in the reclaimed water plume were less than 50% of the drinking water dissolved organic concentrations from which the reclaimed water originated. PMID- 11436801 TI - The Virginia Pipeline: Australia's largest water recycling project. AB - The need to conserve, reuse and recycle water is becoming increasingly important for both environmental and economic reasons. The Virginia Pipeline is Australia's largest water recycling project. More than half the output from (the capital of South Australia) Adelaide's largest wastewater treatment plant is further treated to achieve a product water quality fit for irrigation of vegetable crops with minimal public health restrictions. The project partners have a vision to achieve total reuse. To achieve this vision, recycled water will need to be stored during cool weather periods when the demand for water is low. Temporary storage of this water in poor quality aquifers is the subject of a major research project. PMID- 11436802 TI - Wastewater reuse in Italy. AB - In many parts of Italy, particularly in the South, it has become ever more difficult to meet the water demand. The recent years of drought and the constant increase of water demand for the civil sector have made irrigation supply more problematic. Wastewater reuse could represent a viable solution to meet water demand. The focus of this paper is on the regulation problems, hampering the development of wastewater reuse for irrigation, and on the potentials for reuse, particularly in Southern Italy. Planned exploitation of municipal wastewater could help meeting the irrigation water demand particularly in Southern Italy, where farmers have been practising uncontrolled wastewater reuse for a long time. In Northern and Central Italy, where available water resources generally meet water needs for different purposes, wastewater reuse could play an important role in controlling the pollution of water bodies. Despite the fact that Italian legislation is extremely strict and outdated, for several years in some regions, such as Sicily, wastewater reuse systems have been in operation; furthermore, several projects of wastewater reuse are currently in progress. PMID- 11436803 TI - Reclaimed municipal wastewater--a potential water resource in China. AB - Due to water resource shortage and socio-economic development within twenty years, China faces serious problems of water supply and water pollution. Several criteria and suitable reclamation processes related to water reuse have been created in China, which are helpful to improve the situation of water scarcity. In the future, reclaimed municipal wastewater reuse will mainly be developed for urban and industrial use. Potential supply quantity of reclaimed water, quality of reclaimed water, and reclamation cost are favorable to potential reuses. Based on further public environmental education, on a relevant development of national and local standards for reclaimed water quality, and on an increase of sanitary rate, more and more planned reclaimed water reuse projects would be expected in China. PMID- 11436804 TI - Strategies for development of industrial wastewater reuse in Thailand. AB - Majority of the industrial activities in Thailand are concentrated around Bangkok Metropolitan Area. The ever increasing industrial activities have led to over exploitation of water resources and discharge of significant pollution load. Therefore, it is important to identify the wastewater reuse potentials and develop strategies for its promotion within the industrial sector. Although technological advances have made it possible to treat effluents for industrial re use, in practice, the Thai industries do lack in implementation of such technologies. Promotion of cleaner production concepts and advanced new technologies such as membrane technologies could assist the industry for the implementation of wastewater reuse projects in Thailand. This paper discusses various technical, institutional and management related issues to promote industrial wastewater reuse, with few case studies. PMID- 11436805 TI - Technical-economic modelling of integrated water management: wastewater reuse in a French island. AB - An integrated technical-economic model is used to address water management issues in the French island of Noirmoutier. The model simulates potable water production and supply, potable and non potable water demand and consumption, wastewater collection, treatment and disposal, water storage, transportation and reuse. A variety of water management scenarios is assessed through technical, economic and environmental evaluation. The scenarios include wastewater reclamation and reuse for agricultural and landscape irrigation as well as domestic non potable application, desalination of seawater and brackish groundwater for potable water supply. The study shows that, in Noirmoutier, wastewater reclamation and reuse for crop irrigation is the most cost-effective solution to the lack of water resources and the protection of sensitive environment. Some water management projects which are regarded as having less economic benefit in the short-term may become competitive in the future, as a result of tightened environmental policy, changed public attitudes and advanced water treatment technologies. The model provides an appropriate tool for water resources planning and management. PMID- 11436806 TI - Some economic considerations on wastewater reclamation for irrigation, with reference to the Italian situation. AB - The purpose of this work is to check construction and operation costs for simplified post-treatment trains able to produce water displaying a quality suitable for different agricultural uses. Having in mind the quality of most of surface waters in Italy, the main parameters to control for a general purpose reclaimed water supply are microbial quality and TSS content. Therefore a contact filtration followed by disinfection is to be considered the minimum option, while the use of polishing ponds can be strictly limited in many Italian regions due to the area requirements and to the need to provide a full lining of bottom and levees. Some data referring to surface waters quality and to the Italian epidemiological situation are given. On the basis of the current Italian situation, the construction and operation cost increases due to a polishing section, downstream a standard municipal WWTP, are evaluated and the impact of different technical choices is shortly discussed (disinfection options, chemicals, etc.). Some considerations about the link existing in Italy between the cost of water for irrigation and the agricultural economy are made. PMID- 11436807 TI - Small scale water recycling systems--risk assessment and modelling. AB - This paper aims to use quantitative risk analysis, risk modelling and simulation modelling tools to assess the performance of a proprietary single house grey water recycling system. A preliminary Hazard and Operability study (HAZOP) identified the main hazards, both health related and economic, associated with installing the recycling system in a domestic environment. The health related consequences of system failure were associated with the presence of increased concentrations of micro-organisms at the point of use, due to failure of the disinfection system and/or the pump. The risk model was used to assess the increase in the probability of infection for a particular genus of micro organism, Salmonella spp, during disinfection failure. The increase in the number of cases of infection above a base rate rose from 0.001% during normal operation, to 4% for a recycling system with no disinfection. The simulation model was used to examine the possible effects of pump failure. The model indicated that the anaerobic COD release rate in the system storage tank increases over time and dissolved oxygen decreases during this failure mode. These conditions are likely to result in odour problems. PMID- 11436808 TI - Prospects, problems and pitfalls of urban water reuse: a case study. AB - This paper presents a successful water reclamation and reuse project in the San Francisco Bay area. The project, which includes a water reclamation facility and a separate distribution system, is operated by a wastewater utility and reclaims approximately 4% of its dry-weather flow. Project history, its design and implementation are further discussed. Planning, and especially demand analysis, was critical for project development. Earlier attempts of water reuse were not successful because reclaimed water quality did not match the requirements of potential large industrial customers. Current customers are a mix of public, commercial and residential users who apply the reclaimed water solely for landscape irrigation. In addition, a large fraction of the reclaimed water is used internally in the main wastewater treatment plant. Early connection of largest customers, innovative collaboration with a neighboring reclamation project and cooperation of the local water supplier were very important for project success. Distribution of internal process water consumes most energy. The second major energy use is for the treatment of reclaimed water while distribution of reclaimed water to external customers requires least energy. PMID- 11436809 TI - A methodology to evaluate water and wastewater treatment plant reliability. AB - Evaluating the reliability of treatment processes and treatment facilities should be an important part of the planning and design process for water resource, wastewater treatment, and particularly wastewater reuse projects. With the recent developments in technology, particularly the development of membrane processes and alternative disinfection processes for water and wastewater treatment, there is an increasing need for a common methodology to evaluate the reliability of alternative processes and treatment facilities that utilize different combinations of those processes. To assess the reliability of a treatment facility, several aspects of treatment must be considered including a methodical evaluation of both mechanical reliability and plant performance. A straightforward method for conducting these types of analyses is described herein along with a description of applications of this methodology. A discussion is provided highlighting the value of such a methodology for both the water quality engineer and the risk manager. PMID- 11436810 TI - Dredging through the remains. PMID- 11436811 TI - New UK fisheries conservation measures. PMID- 11436812 TI - Opposition to release of G.M. salmon. PMID- 11436813 TI - Osprey revival from DDT complete in Chesapeake Bay. PMID- 11436814 TI - Historical analysis, a valuable tool in community-based environmental protection. PMID- 11436815 TI - Evolution of metal pollution in the golden horn (Turkey) sediments between 1912 and 1987. AB - The Golden Horn is a heavily polluted water body in a large metropolitan area with a population of approximately 10 million. A 3-m long undisturbed core sample was collected in the Golden Horn, from research vessel RV Knorr, during the third leg of the joint Turkish--American Black Sea expedition in 1989. The core was sliced and dated using the 210Pb isotope technique. The bottom of the core corresponds to the year 1912. Each slice was analysed for major, minor and trace elements by inductively coupled plasma emission spectrometry (ICP). The masses of the measured elements can account for approximately half of the sediment mass. The lithophilic elements Li, K, Rb, Mg, Ca, Ba, Al, La, Ti, V, Mn, Fe, Co and Ni account for more than 90% of the elemental mass and do not show any change in their concentrations between 1912 and 1987. Although anthropogenic elements Mo, Zn, Cr, Cu, Ag, and Cd account for a minute fraction of the elemental mass, their concentrations increase along the core, signifying human influence on chemical composition of the Golden Horn Sediments. Lead was enriched at the bottom of the core suggesting pollution of Golden Horn sediments by this element even at the beginning of the century, but observed concentrations of the remaining anthropogenic elements, at the bottom of the core, can be explained by sedimentary material. Concentrations of pollution-derived elements do not change significantly between 1912 and 1950, but their concentrations increase sharply in the second half of the century. A factor analysis applied to the data set has shown that the inorganic fraction of the Golden Horn sediments includes crustal, marine and two anthropogenic components. One of the anthropogenic components is attributed to the discharges from an iron and steel plant. The second anthropogenic component, which accounted for a larger fraction of system variance, is due to discharges from industries, particularly metalwork plants. PMID- 11436816 TI - Izmit Bay (Turkey) ecosystem after Marmara earthquake and subsequent refinery fire: the long-term data. AB - As a part of Marmara Sea, Izmit Bay (Turkey) has been one of the most polluted sites in the region for the last 25 years. On 17 August 1999, a powerful earthquake along the North Anatolian Fault struck the eastern part of the Marmara region including Izmit Bay. The earthquake destroyed many coastal cities. The Bay was also affected by the quake and subsequent fire in the refinery situated on the north-eastern coast of the Bay. Oceanographic characteristics and polycyclic aromatic hydrocarbon (PAH) levels of Izmit Bay (Marmara Sea) have been investigated to find out the degree of contamination. Seawater samples were collected at nine stations of the Bay in April and September 1999 and the results were compared with those obtained in the previous years (1984 and 1994). Monitoring data are presented for plant nutrients (nitrate + nitrate, ortho phosphate and silicate), dissolved oxygen and chlorophyll a. Surface sediments and mussels, Mytilus galloprovincialis, have been analysed for total PAH (T-PAH) contents in April and September 1999 (before and after the Marmara Earthquake) for the samples collected from eight coastal stations of the Bay. Biomarker (Lysosomal stability and feeding rate) studies at three different sites of the Bay have also been performed to investigate the effect of pollution on mussels. Nitrate + nitrite levels in the upper layer of the eastern part of the Bay increased significantly compared to those measured before the earthquake. Of the samples analysed, the highest o-phosphate concentrations were found in September 1999 in the bottom waters of the Bay. The concentration of chlorophyll a reached its minimum value of the last 15 years. Dissolved oxygen decreased dramatically from 1984 to 1999. Total PAH concentrations measured in April 1999 at both offshore and coastal sites of the Bay were more or less the same (2 micrograms l 1). The subsequent fire after the earthquake caused an increase in the total PAH levels in water column, in sediment and in mussels. Seawater total PAH concentrations ranged between 3.5 and 11 micrograms l-1 at open coast stations and 5-17.5 micrograms l-1 at coastal stations in September 1999. A 2- to 3-fold increase in sediment PAH concentrations (200-5220 mg kg-1 dry weight) was detected after the earthquake. This increase was much more significant in the sediments located around the refinery. More contaminated mussels were detected around the refinery area (110-170 mg kg-1 dry weight). Overall, sediment and mussel PAH concentrations in Izmit Bay are much higher than those found in the other marine systems. In general, the feeding rate and the neutral red retention times of the mussels decreased in some sites of the Bay after the earthquake, but no direct correlation could be detected between the body burden of mussels and biomarkers or between the two biomarker techniques. PMID- 11436817 TI - Occurrence and effects of organotins on adult common whelk (Buccinum undatum) (Mollusca, Gastropoda) in harbours and in a simulated dredging situation. AB - Transplanted common whelk (Buccinum undatum) (Mollusca, Gastropoda) accumulated fairly high levels of organotins (tributyltin (TBT) and triphenyltin (TPT)) during exposure in three harbours with different TBT contamination. This did not though lead to an increase in imposex frequency in the adult females studied. Simulating harbour dredging in an experiment using suspended sediment from one of the harbours only resulted in a low concentration of TBT in the tissues of the common whelk and subsequently no changes in the occurrence and degree of imposex. The common whelk seemed to receive the main part of TBT from the water column and the limited bioaccumulation in the experiment indicates that desorption of TBT from the suspended sediment was slow. After TBT has been totally banned, dredging of contaminated sediments will cause increased exposure of the biota to TBT. Due to slow desorption the increase may however, be slight and temporary to pelagic and epibenthic species unless the settled particles are resuspended. PMID- 11436818 TI - PCBs and organochlorine pesticide residues in eggs of Audouin's gull (Larus audouinii) in the north-eastern Mediterranean. AB - Levels of eight PCB congeners (IUPAC 8, 20, 28, 52, 101, 118, 138, 180) and 13 organochlorine pesticides (alpha-BHC, beta-BHC, lindane (gamma-BHC), heptachlor, heptachlor epoxide, aldrin, dieldrin, endrin, 2,4'-DDT, 2,4'-DDD, 4,4'-DDT, 4,4' DDD, 4,4'-DDE) were measured in unhatched Audouin gull eggs from Aegean Sea (northeastern Mediterranean) colonies in 1997 and 1998. Levels of more persistent congeners 138, 180 and in some colonies 118, predominated among PCBs. Between years significant differences were found in the levels of a variety of contaminants in the same regional colonies (Lipsos, Agathonisi, and Fourni) attributable to temporal changes in diet. Within years differences were limited (1997: PCB congeners 52, 118, 180, heptachlor and 4,4'-DDD; 1998: PCB congener 8 and heptachlor epoxide) which in combination with cluster analysis suggest a temporal rather than spatial pattern of pollution in the Aegean Sea. Maximum median levels of five PCB congeners, total PCBs and eight organochlorine pesticides were found in the Agathonisi and Fourni colonies suggesting an elevated nearby pollution probably from the polluted Menderez River. Levels of all contaminants were too low to have any adverse reproductive effects on the Audouin's gull of the Aegean colonies. PMID- 11436819 TI - Metal levels in seston and marine fish flesh near industrial and metropolitan centres in South Australia. AB - Port Pirie is the site of the largest lead smelter in the world, depositing 250 t of zinc, and 100 t of lead annually into Spencer Gulf. Barker Inlet is adjacent to metropolitan Adelaide, and receives unknown quantities of urban and industrial discharges. Both areas are sites of major commercial and recreational fisheries, contained within delicately balanced marine wetland ecosystems, comprising large areas of mangrove and seagrass habitats. Aldrichetta forsteri and Sillago schomburgkii are major species within these fisheries and as estuarine-dependent species were chosen for this study as indicator species for the detection and monitoring of pollutant impacts in the nearshore marine ecosystems of South Australia. Seston sediment collectors were deployed at each site and analysed seasonally for the presence of cadmium, lead and copper. Flesh samples from A. forsteri and S. schomburgkii were examined seasonally for the presence of cadmium, lead and copper and the results correlated with levels found in the seston sediment at each site. Metal concentrations were also correlated with a biomarker of genotoxicity measured in the same animals (micronuclei in erythrocytes) that were reported previously. Seston levels of cadmium, lead and copper were highest at Port Pirie, followed by Barker Inlet and were lowest at Wills Creek, with cadmium undetectable at the latter site. Metals in seston varied considerably with season, with generally higher levels in winter samples. In fish flesh, metal levels followed broadly similar trends as for seston. Spearman rank correlations between metals in seston and in flesh were strongly positive. There was also a significant correlation between flesh concentrations of each metal and the frequency of micronuclei in erythrocytes. This study has shown that seston concentration of pollutant metals are high in areas of industrial activity, and that these levels are also reflected in metal content of fish flesh. Mean flesh levels of cadmium and copper did not exceed Australian health based maximum permitted levels of fish for human consumption, whereas flesh levels of lead in fish from Port Pirie and Barker Inlet exceeded these standards in each of the seasons monitored. This may represent a significant dietary source of lead in humans, especially at Port Pirie where human lead exposure from terrestrial sources is important. There may also be the potential for accumulation of metals in residents of metropolitan Adelaide whose diets are high in fish (and/or crustaceans), particularly estuarine-dependent species, such as A. forsteri and S. schomburgkii. The study also showed that a non-specific biomarker of genotoxicity (micronuclei in erythrocytes) is potentially useful as a monitoring technique in fish species to evaluate their exposure and genotoxic responses to pollutants in South Australian waters. These data represent a snapshot of the current situation in this area and may act as background levels against which future improvements or decrements in water quality may be compared. PMID- 11436820 TI - Trace element studies in Laucala Bay, Suva, Fiji. AB - The trace metal distribution in the sediments of Laucala Bay, a coastal lagoon with a barrier reef and significant freshwater input, adjacent to Suva, the capital of Fiji (150,000 people), was studied from the point of view of assessing the significance of anthropogenic sources relative to natural ones. Surface sediments from 25 sites in the Bay were analysed for particle size distribution, organic carbon and major (Si, Al, Fe, Ca, Mg) and trace metal (Mn, Zn, Cu, Pb, Cd, Hg) contents. Suspended sediments from one site and shellfish samples from five sites were also analysed. The sediments were found to be mostly of terrigenous origin. Particle size distribution, organic carbon and major elemental composition of the sediments were generally related to location within the Bay. A significant natural source of trace metals in the sediments is the suspended solids transported into the Bay by rivers. The spatial distribution of trace metals could not be fully explained by the particle size distribution and mineral composition of the sediments. Contributions from anthropogenic sources were also suspected, but the degree of enrichment from such sources is not great at the present time. PMID- 11436821 TI - Brown tides and mariculture in Saldanha Bay, South Africa. AB - In 1997, the brown tide organism, Aureococcus anophageffens, was detected for the first time in Saldanha Bay, South Africa. Its presence was limited to an isolated, tidal dam that was similarly impacted during the late summer of the following two years but not in 2000. Bloom concentrations are typically of the order of 10(-9) cells l-1. This is one of the few reported occurrences of these nuisance blooms outside the north-eastern United States. A small oyster grow-out facility based in the dam has been severely affected by the reduced growth of oysters during these blooms. Reduced flushing of this culture site is a possible explanation for bloom initiation and persistence. However, Aureococcus blooms can be considerably more extensive as was evident during 1998 when the whole of the bay system, including Langebaan Lagoon, was affected for 6-8 weeks during late summer. PMID- 11436822 TI - Contamination of the coastal waters of Bermuda by organotins and the triazine herbicide Irgarol 1051. AB - A study of the distribution of the 'booster' biocide 2-methylthio-4-tert butylamino-6-cyclopropyl amino-s-triazine (Irgarol 1051) was carried out in the coastal waters of Bermuda. Irgarol 1051 concentrations (as determined by GC/MS) up to 590 ng l-1 have been measured within Hamilton Harbour. The data presented herein unequivocally demonstrate contamination of the coastal system of Bermuda by Irgarol 1051. Concurrently, TBT concentrations were measured and results indicate that levels are falling through legislated changes in antifouling treatments, from 220 ng l-1 in 1990 to < 20 ng l-1 (as Sn) by 1995, in the open water area of Hamilton Harbour. Concentrations of TBT immediately offshore from a boatyard were found to be > 600 ng l-1 (Sn), indicating continuing release due to painting operations and sediments in the area. PMID- 11436823 TI - Mercury and silver concentrations in sediments from the Port of Ensenada, Baja California, Mexico. PMID- 11436824 TI - A food of one's own. PMID- 11436825 TI - Still not out of the woods. PMID- 11436826 TI - If you believe embryos are humans.... ...then curbing research on stem cells is an odd place to start protecting them. PMID- 11436827 TI - Web alert. PMID- 11436828 TI - Web alert. PMID- 11436829 TI - [Changes in bacterial flora of nasopharynx in children treated with long-term administration of macrolides]. PMID- 11436830 TI - [Roxithromycin therapy of patients with mycobacterium avium complex lung infection]. PMID- 11436831 TI - [Effect of clarithromycin on monocyte-derived inflammatory cytokine production in bacterial infections with special reference to Mycobacterium avium infection]. PMID- 11436832 TI - [ Selective inactivation of Pseudomonas-derived aminoglycoside 3' phosphotransferase by macrolides]. PMID- 11436833 TI - [Erythromycin uptake pathways in Shigella]. PMID- 11436834 TI - [Changes in bacterial flora after long-term administration of clarithromycin]. PMID- 11436835 TI - Society for Experimental Biology. Annual meeting, 2nd-6th April 2001, The University of Kent (Canterbury). Plant and cell biology abstracts. PMID- 11436836 TI - The journey back. Part two: the test. PMID- 11436837 TI - Digital hearing aids. PMID- 11436838 TI - Physicians destined to benefit from new Web applications and sites. PMID- 11436839 TI - 2001 Scientific Meeting of the Research Society on Alcoholism. June 22-28, 2001, Montreal, Quebec, Canada. Abstracts. PMID- 11436841 TI - 24th annual conference on shock. Saturday, June 9 to Tuesday, June 12, 2001, Marco Island, Florida. Abstracts. PMID- 11436840 TI - 36th meeting of the Canadian Congress of Neurological Sciences. Halifax, Nova Scotia, June 12-16, 2001. Abstracts. PMID- 11436842 TI - Knowledge of fraud among biostatisticians. PMID- 11436843 TI - Using permuted blocks to register and randomize patients in clinical trials. PMID- 11436845 TI - Prostate-specific antigen in prenatal screening for Down's syndrome. PMID- 11436846 TI - Elective surgery boosts cerebral palsy risk. PMID- 11436848 TI - Physiological Society of Japan. Proceedings of the 77th annual meeting. March 27 29, 2000, Tokyo. Abstracts. PMID- 11436847 TI - Caribbean Health Research Council. 46th annual council and scientific meetings. April 25-28, 2001, Jamaica. Abstracts. PMID- 11436849 TI - Abstracts from the 4th annual meeting of the American Society of Gene Therapy. May 30-June 3, 2001, Seattle, Washington. PMID- 11436850 TI - Pediatric Endocrinology Montreal 2001. Friday, July 6-Tuesday, July 10, 2001, Montreal, Quebec, Canada. Abstracts. PMID- 11436851 TI - 6th International Congress of Vertebrate Morphology. Jena, Germany, July 21-26, 2001. Abstracts. PMID- 11436852 TI - Neurosteroids on our minds. PMID- 11436853 TI - Can simulations be too dry? PMID- 11436854 TI - The perils of using the human genome sequence: lessons from CALCRL. PMID- 11436855 TI - Pulsed electromagnetic field therapy in the management of knee OA. PMID- 11436856 TI - Treatment of women with aPL in pregnancy. PMID- 11436857 TI - Caudal epidural injections. PMID- 11436858 TI - Interleukin 1 gene polymorphisms. PMID- 11436859 TI - A case of shingles mimicking carpal tunnel syndrome. PMID- 11436860 TI - Azathioprine hypersensitivity. PMID- 11436861 TI - Immunosuppressive treatment for vasculitis associated with Lyme borreliosis. PMID- 11436862 TI - Bilateral transient osteoporosis of the knee in pregnancy. PMID- 11436863 TI - Anticholesterol antibody levels in patients with systemic lupus erythematosus. PMID- 11436864 TI - Intravenous immunoglobulin for treatment of gastro-intestinal haemorrhage in dermatomyositis. PMID- 11436865 TI - Sjogren's syndrome: an unusual cause of Bell's palsy. PMID- 11436866 TI - Alpha1 antitrypsin phenotypic variability is not associated with ANCA in southern Chinese. PMID- 11436867 TI - A case of cholesterol embolism with ANCA treated with corticosteroid and cyclophosphamide. PMID- 11436868 TI - HLA-DRB1*0301 and DQA1*0501 in RA. PMID- 11436869 TI - Sixty-five pieces of silver. PMID- 11436870 TI - Vox populi. PMID- 11436871 TI - Signifying what? PMID- 11436872 TI - Who is there to listen? PMID- 11436873 TI - A new year--a new era. PMID- 11436874 TI - "The judgment is reversed". PMID- 11436875 TI - [Parry-Romberg syndrome with glaucoma and pregnancy]. PMID- 11436877 TI - Second trimester pregnancy termination in primigravidas by double application of dinoprostone gel and intramuscular administration of carboprost tromethamine. AB - A randomized study of legal pregnancy termination in the second trimester has been performed in primigravidas by endocervical application of Prepidil gel (dinoprostone gel) in combination with intramuscular application of Prostin 15 M (carboprost tromethamine). The study comprised 97 patients. The control group (N = 35) included patients in whom pharmacologic preparation of the cervix was performed by a singleton endocervical application of 0.5 mg dinoprostone gel and intramuscular administration of Prostin 15 M (Carboprost Tromethamine) (1 ml/2 h) 10 h later and until abortion. The investigated group A (N = 31) underwent pharmacologic preparation of the cervix by endocervical application of 0.5 dinoprostone gel, 2 times during an interval of 4 hours. Intramuscular injection of Prostin 15 M (Carboprost Tromethamine) (1 ml/h) was administered 6 hours after the second application of gel and until abortion. The investigated group B (N = 31) underwent pharmacologic preparation of the cervix by a singleton double-dose endocervical application of 0.5 mg dinoprostone gel and intramuscular treatment by 1 ml/2 h of Prostin 15 M (Carboprost Tromethamine) applied 10 hours later and until abortion. On the basis of obtained results, a conclusion was made that the NEW combination of endocervical and intramuscular application of prostaglandins gives statistically significantly shortened abortion interval (the control group x = 9.97; SD = 5.22 versus investigation group: A-x = 6.80; SD = 4.31 and investigation group: B-x = 5.55; SD = 3.48). Better pharmacologic preparation of the uterine cervix resulted in shorter abortion interval and decreased amount of systemically applied prostaglandins. A shortened abortion interval significantly decreases the rate of immediate, early and late complications. A decreased amount of systemically applied prostaglandins results in significantly lower rate of unwanted side-effects caused by prostaglandins, making this method of pregnancy termination significantly cheaper. PMID- 11436878 TI - The impact of biomedical literature published in the province of Vojvodina on researchers in the world and in Yugoslavia. AB - This study uses bibliometric analysis to evaluate scientific biomedical literature published on the territory of Vojvodina and its impact on researchers in Yugoslavia and in the world. The study sample comprised 6.979 publications from the territory of Vojvodina published in the period 1986-1997. The following parameters were determined: productivity of authors and institutions where they were employed and some other bibliometric parameters. The impact of these publications on other researches was analyzed for the same period, by citation analysis of papers published in 3 most eminent medical journals in Yugoslavia (3.440 articles, 58.484 references) and in Science Citation Index. Results of the study revealed that production of biomedical literature on the territory of Vojvodina was at high level. Medical research presented in journals of Vojvodina, which are the carriers of current information and accomplishments in science, technique, and practice in biomedicine, was satisfactory. Biomedical articles published in Vojvodina showed a tendency towards increase in number of authors, whereas the greater number of publications were written by a small number of extremely productive authors and institutions. Biomedical researches in Yugoslavia usually cite only foreign literature, while domestic references are mostly self-citations or citations of older literature. The impact of publications published in Vojvodina on other researchers in Yugoslavia is evident, but it is greatest on the territory where they are published. In regard to biomedical journals from Vojvodina "Medical review" is the most cited journal in Yugoslavia and in SCI. Most of the cited references belong to a small number of authors. Thus, according to bibliometric criteria the impact of medical science and professional practice in Vojvodina on international scientific streems is negligible, as well as the impact of papers published in Vojvodina on researchers in the world. Only 0.4 works published in Vojvodina, 0.87% papers from journals published in Vojvodina and written by 0.59% authors were cited in SCI. PMID- 11436879 TI - Holistic vets open to any research programs--but lack funding. PMID- 11436880 TI - Abstracts of the IX Congress of the International Society for Peritoneal Dialysis. June 26-29, 2001, Montreal, Canada. PMID- 11436881 TI - Abstracts of the 36th annual meeting of the European Association for the Study of the Liver. April 18-22, 2001, Prague, Czech Republic. PMID- 11436882 TI - Risks and benefits in pediatric CT. MR/CT Committee of the Society of Pediatric Radiology. PMID- 11436883 TI - Fallout from recent articles on radiation dose and pediatric CT. PMID- 11436884 TI - Sonography of brain tumors in infants and young children. AB - The sonographic features of five brain tumors are presented to emphasize the variability of imaging findings and the role that sonography may play in the initial diagnosis, determination of tumor vascularity, and biopsy guidance. PMID- 11436885 TI - Diagnostic and interventional ultrasonography in neonatal and infant lumbar puncture. AB - BACKGROUND: Lumbar puncture (LP) may be unsuccessful clinically, prompting image guided LP by radiologists. Objective. To investigate the utility of ultrasound (US) in diagnosing the cause of failed LP and in guiding LP. MATERIALS AND METHODS: Neonates and infants referred for image-guided LP underwent spine US of the thecal sac. When indicated, image-guided LP was performed. RESULTS: Forty seven evaluations and interventions were performed in 32 patients. All patients were initially evaluated after failed blind LP attempts. Twenty-three of the initial US studies showed intrathecal and/ or epidural echogenic hematoma, which obliterated the CSF space; 5 showed minimal fluid, and 4 had normal examinations. LP was deferred or cancelled in 14 cases based upon initial US findings. Image guided LP was performed 32 times in 19 patients. US guidance was used in 26, fluoroscopy in 3, and fluoroscopy with US assistance in 3. Using US, LP was performed in 9 patients with no visible CSF: 2 samples were sufficient for culture only. Six patients had minimal CSF US: 4 provided usable CSF samples. Clear CSF space was seen in 11: all had successful LP. CONCLUSIONS: US can disclose the cause of failed LP, can help determine whether or not to intervene further, and can provide guidance for LP. PMID- 11436886 TI - Desmoplastic infantile ganglioglioma (DIG): cranial ultrasound findings. AB - Desmoplastic infantile ganglioglioma (DIG) is a rare brain tumor encountered in infants. In spite of its large size at presentation and occasional high mitotic activity on histopathology, the tumor has a good prognosis. A 7-month-old baby girl developed increasing head circumference. On ultrasound, a large multicystic mass was seen. We report the cranial ultrasound findings for the first time. Correlative imaging of this recently recognized entity is shown. PMID- 11436887 TI - Mediastinal interfaces and lines in children: radiographic-CT correlation. AB - BACKGROUND: Various mediastinal interfaces and lines are well known in adults, but not fully understood in children. OBJECTIVE: To review the mediastinal interfaces and lines on plain radiographs in the paediatric age group with CT correlation. MATERIALS AND METHODS: Soft copies of 195 sets of CT examinations and concurrent chest radiographs in 180 paediatric patients (age 1 month to 15 years) were retrospectively reviewed. The frequency of visualisation and the anatomical basis of the mediastinal interfaces and lines were assessed. The evaluated mediastinal interfaces and lines were superior vena cava interface, descending aorta interface, left paraspinal interface, azygoesophageal recess interface, right paratracheal stripe, anterior junction line and posterior junction line. RESULTS: Chest radiographs showed the SVC interface in 161, the descending aorta interface in 155, the left paraspinal interface in 98, the azygoesophageal recess in 94, the right paratracheal stripe in 53, the anterior junction line in 15 and the posterior junction line in 10. Non-visualisation of mediastinal interfaces and lines on plain radiographs was explained by normal anatomical difference compared with the adult and underlying intrathoracic abnormalities when they were correlated with CT. The frequency of visualisation of the SVC interface, descending aortic interface, left paraspinal interface and azygoesophageal recess fluctuated with age, while the frequency of the right paratracheal stripe, anterior junction line and posterior junction line increased with age. CONCLUSIONS: Awareness of the frequency of visualisation and the anatomical basis of the mediastinal interfaces and lines in paediatric patients may be helpful for interpretation of chest radiographs. PMID- 11436888 TI - Technique and clinical applications of full-inflation and end-exhalation controlled-ventilation chest CT in infants and young children. AB - BACKGROUND: The inability of young children to cooperate with breath holding limits the usefulness of chest CT. OBJECTIVE: To describe the technique and utility of a non-invasive method called controlled-ventilation CT (CVCT) for obtaining motion-free full-inflation and end-exhalation images of the lung in infants and young children. MATERIALS AND METHODS: Eighty-seven children (ages 1 week to 5 years, mean 2 years) underwent CVCT of the chest during suspended respiration at full-lung inflation and end-exhalation for a variety of clinical indications. Respiratory pauses were produced using conscious sedation and positive-pressure face-mask ventilation. Forty-one of 87 children had recordings of respiratory motion during CVCT. RESULTS: Respiratory pause lengths increased with age (P < 0.003), were highly reproducible (r = 0.85), and lasted sufficiently long to be practical for full-inflation (24 +/- 9 s) and end exhalation (12 +/- 5 s) CT scanning. Full-inflation CVCT was useful in evaluating tracheal and bronchial stenosis, bronchial wall thickening, early bronchiectasis, bronchial fistula, extent of interstitial fibrosis, and lung nodules. End exhalation CVCT was useful in evaluating tracheomalacia and air trapping. CONCLUSION: Controlled-ventilation chest CT is a practical and reliable technique that promises to be clinically useful for a number of clinical indications in infants and young children. PMID- 11436889 TI - Swallowing dysfunction in infants less than 1 year of age. AB - BACKGROUND: Gastroesophageal reflux (GER) occurs frequently in infants and children and is implicated as a trigger for reactive airways disease. While evaluating patients for GER by upper gastrointestinal studies (UGI), we frequently noticed laryngeal penetration or tracheal aspiration in infants < 1 year of age. We conducted this prospective study to assess the incidence of swallowing dysfunction in infants with vomiting or respiratory symptoms. METHODS: Between February 1994 and August 1997, 1,003 UGI in infants < 1 year of age were performed by two experienced pediatric radiologists. Fluoroscopy of swallowing using an appropriate image intensifier was observed as part of the UGI in all patients. In addition to evaluating premature infants, former premature infants, and infants with underlying conditions such as bronchopulmonary dysplasia (BPD), congenital heart disease (CHD), esophageal atresia and/or tracheoesophageal fistula (EA), and neurologic disorders, we assessed swallowing in 472 full-term infants in the general population who had respiratory symptoms or suspected GER, but no other apparent abnormalities. Swallowing was not assessed in patients with congenital bowel obstruction. The presence of normal swallowing or swallowing dysfunction was recorded immediately following the study. Chest radiographs obtained on the same day as the UGI were evaluated in the study. Forty-four videotaped modified barium-swallow studies (MBSS) were obtained in 25 general population infants and reviewed frame-by-frame to determine the length of time that barium could be visualized in the subglottic trachea. RESULTS: The incidence of swallowing dysfunction is significant in premature and former premature infants, those with BPD, CHD, EA, various syndromes, and neurologic abnormality. In the general population of full-term infants referred for evaluation of vomiting or respiratory symptoms, 63 (13.4 % of 472) had swallowing dysfunction. Forty-four had tracheal aspiration (TA) and 19 had laryngeal penetration (LP). Gastroesophageal reflux was found in 79.5 % with TA and in 68.4 % with LP. The MBSS confirmed swallowing dysfunction in all patients shown to have vocal cord penetration or tracheal aspiration by UGI. CONCLUSIONS: Careful evaluation of swallowing during an UGI can identify swallowing dysfunction in full-term infants who have respiratory problems, as well as in infants with an abnormality that predisposes the patient to aspiration. Episodes of tracheal aspiration may be fleeting and overlooked if swallowing is not assessed carefully. An MBSS can confirm the observation of swallowing dysfunction found during an UGI and assist in planning appropriate dietary therapy that minimizes the likelihood of tracheal aspiration during feeding. PMID- 11436890 TI - Radiologic considerations in patients undergoing the Nuss procedure for correction of pectus excavatum. AB - The Nuss procedure has succeeded in minimizing incisions, blood loss, sternal fracturing, operating room time, recovery time, and length of hospital stay. Knowledge of the pre- and postoperative radiologic considerations is essential in providing appropriate imaging support to the surgeons performing this innovative, minimally invasive procedure. PMID- 11436891 TI - Magnetic resonance imaging of the fetus in congenital intrathoracic disorders: preliminary observations. AB - BACKGROUND AND OBJECTIVE: Advances in magnetic resonance imaging (MRI) provide high-quality images of the intrathoracic organs. We studied the ability of MRI to define spatial relationships of the fetal lungs and measured lung volume in two cases of congenital diaphragmatic hernia (CDH), one of severe oligohydramnios secondary to bilateral cystic renal dysplasia and one case of prenatal chylothorax. PATIENTS AND METHODS: We performed pelvic MRI using single-shot fast spin echo (SSFSE) pulse sequence in four pregnant women referred because of abnormal prenatal ultrasound (US) findings associated with pulmonary hypoplasia. RESULTS: The exact anatomic position of the contents of the hernia in CDH, including the position of the liver, was better defined with MRI. Pleural effusions were identified as well as the renal abnormality in the case of oligohydramnios. Lung volume was measured and the degree of pulmonary hypoplasia was quantified in every case. Lung-to-thorax ratio was calculated in the case of fetal chylothorax. CONCLUSION: Ongoing work suggests that MRI can provide additional detailed quantitative information in prenatal disorders associated with fetal lung compression and resulting hypoplasia. Correlation of fetal lung volume with postnatal management and outcome may affect prognosis in these cases. PMID- 11436892 TI - Traumatic prenatal sigmoid perforation due to amniocentesis. AB - A variety of fetal injuries, including those inflicted to the gastrointestinal tract by amniocentesis, have been reported before. This brief report describes the first documented case of sigmoid perforation owing to the common procedure of amniocentesis that manifested as abdominal distention at birth. A potential link between this complication and a recent increased incidence of "intrauterine spontaneous perforation" of the gastrointestinal tract has been mentioned. Practicing radiologists are encouraged to inquire directly about the history of amniocentesis in unexplained cases of intrauterine intestinal perforation. PMID- 11436893 TI - Acute diaphragmatic paralysis caused by chest-tube trauma to phrenic nerve. AB - A 3 1/2-year-old child developed unilateral diaphragmatic paralysis after chest drain insertion. Plain chest X-ray demonstrated paravertebral positioning of the chest-tube tip, and magnetic resonance imaging revealed hematomas in the region of the chest-tube tip and the phrenic nerve fibers. The trauma to the phrenic nerve was apparently secondary to malposition of the chest tube. This is a rare complication and has been reported mainly in neonates. Radiologists should notify the treating physicians that the correct position of a chest drain tip is at least 2 cm distant from the vertebrae. PMID- 11436894 TI - Description of a rare type of posterior pelvis traumatic involvement: the green stick fracture of the sarcum. AB - Fractures of the pelvic ring in children behave differently than in adults. In children, bone elastic properties allow for plastic deformation instead of complete breaks. To the best of our knowledge no reports of this bony behavior have been described in the sacrum, and this could explain the presence of single displaced anterior breaks without evident posterior pelvic disruption. The objective of this paper is to document a rare type of posterior pelvis trauma: the green-stick fracture of the sacrum. PMID- 11436895 TI - Fibrous hamartoma of infancy at the wrist and the use of MRI in preoperative planning. AB - Fibrous hamartoma of infancy is an uncommon, self-limiting benign tumour that presents during the first 2 years of life, developing from subcutaneous fibrous tissue proliferation at almost any site. We describe the results of MRI of a lesion at the wrist. The scans enabled a planned surgical resection with the aim of preventing local recurrence without damaging important neurovascular structures. PMID- 11436896 TI - Congenital epulis: prenatal diagnosis by ultrasound. AB - We present a case of congenital epulis, diagnosed prenatally with US. Congenital epulis is a benign gingival tumour whose differential diagnosis includes other oral-facial masses such as haemangioma, granular cell myoblastoma and cystic hygroma. This tumour can cause obstruction of the airway or feeding problems in the newborn child. Surgical excision is the treatment of choice. PMID- 11436897 TI - Jejunoduodenogastric intussusception--a rare complication of gastrostomy tube migration. PMID- 11436898 TI - Small-bowel intussusceptions occurring around nasojejunal enteral tubes--three cases occurring in children. PMID- 11436899 TI - Accidental iron ingestion. PMID- 11436900 TI - Mediastinal and abdominal lymphadenopathy imitating lymphoma in an adolescent with autoimmune hepatitis. PMID- 11436901 TI - Inflammatory pseudotumor of the larynx: MR findings in a child. PMID- 11436902 TI - European Conference on Infectious Diseases and Tropical Medicine. EuCID 2001. 3-6 May 2001, Leipzig. Abstracts. PMID- 11436903 TI - Farewell to a surgical giant. A tribute to C. Walton Lillehei, M.D., Ph.D. PMID- 11436905 TI - Closure of the Thurso veterinary investigation centre. PMID- 11436904 TI - Calculation and prescription of dose for total body irradiation. AB - The use of large total body fields creates a unique set of problems that stress the accuracy of techniques routinely used for dose calculation. This paper discusses an approach suggested by the Children's Cancer Study Group (CCSG) for both prescribing the total body irradiation (TBI) dose and calculating the beam on time or meter set needed to deliver it. It is aimed at guaranteeing the accuracy of the calculations, while at the same time ensuring a high degree of compliance for the various CCSG protocols using TBI. Data supporting the various CCSG recommendations are presented. PMID- 11436906 TI - The emergency medicine department system: a study of the effects of computerization on the quality of medical records. AB - A template based computerized medical record system known as the Emergency Medicine Department System (EMDS) was installed in the emergency department of the National University Hospital, Singapore, replacing handwritten records. A study was carried out to show how the implementation of the EMDS improved the quality of medical records. A retrospective review of old manual records and the ones generated by the EMDS was done by means of a scoring system. The raw scores represent the amount of information captured. The calculated means of scores were then used to compare the records. It was found that EMDS improves the quantity of data capture over the old records in all sections compared. This was seen regardless of the experience of the user. The use of a non-structured generic template results in less data captured compared with a structured symptom specific template. The design of questions has a great influence in that a double choice question captures more data than single-choice questions. Building in 'locking' or enforcement mechanisms in the EMDS also helped achieve almost full capture of critical information, such as examination time. PMID- 11436907 TI - Patients' satisfaction with the staff function in an emergency department. AB - Patients' satisfaction with the functional capacity and attitude of the permanent staff working in the morning hours in the emergency department (ED) of a community hospital was compared with that of the staff working during the evening and night shifts. A total of 285 patients given care in the ED were interviewed according to a 'satisfaction' questionnaire regarding the function and attitude of the ED staff during the morning and evening/night shifts. The mean waiting time until a doctor was seen during the morning shift was 25 +/- 17 minutes for non-hospitalized patients and 25 +/- 8 minutes for the hospitalized ones, whereas during the evening and night hours the waiting times were 22 +/- 17 minutes and 19 +/- 13 minutes respectively. The number of laboratory examinations performed during the evening and night shifts markedly exceeded that carried out during the morning. The mean staying time in the ED for both non-hospitalized and hospitalized patients during the morning was by 23% shorter than that during the evening and night shifts. The patients expressed their overall satisfaction with the ED staff in both shifts with high evaluation marks. It is concluded that the survey indicates that the permanent ED staff during the morning hours are more efficient compared with those working during the evening and night shifts. PMID- 11436908 TI - Update on the development of emergency medicine as a specialty in Turkey. AB - Emergency medical care, both prehospital and hospital-based, is currently provided by general practitioners in over 90% of the emergency departments in Turkey. In the early 1990s, government and university leaders recognized that Turkey needed to improve its emergency medical care system, and they chose to adapt the mature and tested Anglo-American model of emergency medicine (EM). EM was declared to be an independent specialty by the Ministry of Health in 1993. The first paramedic school and the first EM residency programme (36 months in length) were opened at the Dokuz Eylul University in 1993 and 1994, respectively. In 1995, the Emergency Medicine Association of Turkey (EMAT) was established. Today, there are 14 EM residency programmes around the country, and these are trying to design a common curriculum. The connection between departments is improving with annual meetings organized by EMAT. In addition, EMAT is developing international collaboration in the Middle East region. The Turkish government is trying to promote EM specialist physicians and paramedics in the national emergency care system. PMID- 11436909 TI - Narcotic analgesia in the acute abdomen--a review of prospective trials. AB - Withholding administration of narcotic analgesia in patients with acute abdominal pain for fear of masking pathology is still pervasive in current medical practice. We reviewed all the prospective trials that investigated the safety, adverse affects, and ultimate outcome in patients with acute abdominal pain receiving narcotic analgesia within the emergency department (ED). No adverse outcomes or delays in diagnosis could be attributed to the administration of analgesia. Based on this research, we propose that it is safe and humane to administer narcotic pain relief to patients presenting to the ED with acute abdominal pain provided no contraindications exist. PMID- 11436910 TI - Disaster medicine--reflections on goals and developments. The future of emergency and disaster medicine. PMID- 11436911 TI - Conversion of atrial fibrillation into a sinus rhythm by coronary angioplasty in a patient with acute myocardial infarction. AB - Atrial tachyarrhythmias are important complications occurring in more than 8% of acute myocardial infarctions (AMI). Atrial fibrillation (AFi) during the early phase of AMI is caused by atrial ischaemia, atrial distension due to the left ventricular failure or significant diastolic left ventricular dysfunction. AFi in patients with inferior and posterior AMI indicates at least two vessel coronary diseases, a circumflex coronary artery (CX) occlusion before taking off of the left atrial branches as well as significant stenosis or occlusion of the right coronary artery (RCA). In this article the case of a 67-year-old woman with an acute infero-posterior AMI is described. AMI was complicated with a left heart failure, acute AFi with tachyarrhythmia, transient arterial hypotension and ischaemic mitral regurgitation. Emergency coronary angiography disclosed occlusion of the CX, myocardial infarct related artery, and significant stenoses of the RCA. After opening the occluded CX during the PTCA, AFi with a tachyarrhythmia of 160 beats per minute (bpm) immediately converted into a sinus rhythm with 80 bpm, followed by a normalization of blood pressure and cardiac recompensation. Our case report supports the opinion that AFi in patients with inferior and posterior AMI indicates at least a two-vessel coronary disease. Reopening of the occluded atrial coronary branches during urgent medical treatment was casual and effective treatment of both ischaemic heart disease and consequent AFi. PMID- 11436912 TI - On the look out for a rarity--interstitial/cornual pregnancy. AB - Interstitial (cornual) pregnancy is a rare form of ectopic pregnancy that can be difficult to diagnose both clinically and ultrasonographically. However, early detection is essential to avoid fatal outcome. We report a case of interstitial/cornual ectopic pregnancy. The patient presented with uterine rupture over the cornual region resulting in catastrophic bleeding. The risk factors, clinical features and ultrasonographic signs are reviewed. PMID- 11436913 TI - Pre-cervical soft tissues: a cautionary note. AB - Visualization of pre-cervical soft tissue swelling on a lateral radiograph is an important indicator of possible bony injury in the context of cervical spine trauma. We report a case where apparent swelling was in fact an artefact due to an inappropriately placed endotracheal tube. We recommend review of the position of endotracheal tubes when interpreting pre-cervical soft tissue swelling. PMID- 11436914 TI - Ventricular fibrillation following inhalation of Glade Air Freshener. AB - Intentional hydrocarbon inhalation can be fatal. Death can be secondary to hydrocarbon's cardiopulmonary effects. We present a case of a patient who survived ventricular fibrillation after inhalation of Glade Air Freshener, which contains short chain aliphatic hydrocarbons (butane and isobutane). Unlike our case, myocardial sensitization and hypoxia are more commonly described with aromatic, halogenated or longer chain hydrocarbons. PMID- 11436915 TI - Exertional rhabdomyolysis in a body builder abusing anabolic androgenic steroids. AB - Rhabdomyolysis, or acute skeletal muscle destruction, may be accompanied by myoglobinaemia, myoglobinuria, and an elevated serum creatine kinase level. This disorder has many potential causes. In this article, the authors describe a case of rhabdomyolysis occurring after vigorous weight lifting by a man who was supplementing his weight-training programme with the intake of anabolic androgenic steroids dispensed to him by a colleague. PMID- 11436916 TI - Vitamin deficiency in a toddler reluctant to use her arm. AB - Children frequently present to accident and emergency departments with limb pain, usually following trauma. An unusual cause of atraumatic limb pain in a toddler is described in this report. Rickets should be added to the differential diagnosis of atraumatic limb pain in this age group. The possibility of acute bony injury (following unwitnessed trauma) not apparent on X-ray needs to be considered in this case. Suspicion of rickets, based on clinical features and radiographic findings, can be confirmed by biochemical assays. PMID- 11436917 TI - Bilateral posterior shoulder dislocation: the importance of the axillary radiographic view. AB - Whilst posterior shoulder dislocation is rare, it remains a frequently missed diagnosis. In all patients with a history of seizure, electrocution or similar trauma, where external rotation of the shoulder is limited, we recommend that an axillary or modified axillary view be added to the standard two radiographic shoulder views as routine protocol, to avoid missing a posterior dislocation. PMID- 11436918 TI - Reduction of a posterior shoulder dislocation during Swimmer's view radiography. PMID- 11436919 TI - The specialty of Emergency Medicine in an international perspective. PMID- 11436920 TI - Abdominal stab wounds: a five-year patient review. AB - Over a period of 5 years, 101 patients were treated for abdominal stab wounds at our emergency department. Exploratory laparotomy was performed in 41 of these cases. The indication for laparotomy was set by clinical investigation in 20 cases, instrumental exploration in six patients, ultrasound in six, paracentesis in seven and computerized tomography scanning in two cases. A review of the results of the treatment of these abdominal stab wounds is compared with the literature. Routine laparotomy for this type of penetrating trauma to the abdomen proves to be no longer warranted. Clinical and especially diagnostic studies should be able to select patients in whom laparotomy should be performed. When these studies are unable to exclude severe injury, laparotomy remains more prudent than expectant observation. PMID- 11436921 TI - In search of the most suitable technique for truncal spinal immobilization with associated radiography. AB - The spineboard (SB) and the vacuum mattress (VM) are utilized for prehospital and emergency department (ED) immobilization of the spine. While permitting excellent pictures to be taken, the SB is a very painful device that can only be used for a limited time. The current study investigated the feasibility of different models of the VM for radiography. Computed tomography for each of seven VMs showed different degrees of shadows from the sac containing the polystyrole balls. This was related first of all to a high contrast of the sac in comparison with its contents and to a sometimes considerable shrinkage of the latter that resulted in further folding into the sac and, secondly, also to very broad mattresses, additional chambers within the VM and various grips and supports for lifting the mattress. Therefore, some features designed for prehospital use of the VM are actually acting against its use for diagnostic purposes. The aim of this study was to discuss and identify possible properties of a device that is useful for both immobilization and diagnostic purposes. PMID- 11436922 TI - The yield of blood cultures in a department of emergency medicine. AB - This study sought to determine the yield of blood cultures drawn in the department of emergency medicine. The results of 730 blood cultures taken from 718 patients were retrospectively analysed. The total percentage of positive cultures was 9.7%. Only 3.4% of the blood cultures were classified as true bacteraemia and the rest as contaminants. The commonest type of isolate was coagulase-negative staphylococci (49%), which were considered contaminants in all cases. Other contaminants represented 13.2% of all the positive blood cultures. The following bacteria comprised the group of true bacteraemia: Escherichia coli (12.6%), Streptococcus pneumoniae (9.8%), viridans streptococci (7%), Staphylococcus aureus (2.8%), Bacteroides fragilis (2.8%), Moraxella species (1.4%) and Flavobacterium species (1.4%). Blood cultures were positive in 3.6% of patients with pneumonia and in 10% of patients with urinary tract infections. In patients with fever of unclear source blood cultures were positive in 3.1% of children between 0-36 months of age and in 1.1% of patients older than 16 years. As a whole, patients with positive blood cultures were clinically sicker, a higher percentage of them required admission to the hospital and had higher temperatures or rapidly fatal disease, compared with the group of patients with negative blood cultures. In order to improve the yield of blood cultures in febrile patients, first, better a priori identification of those subjects at high risk for bacteraemia will reduce the number of unnecessary blood cultures and second, sterile venipuncture techniques should be improved in order to reduce the number of contaminants. PMID- 11436923 TI - Prospective evaluation of adenosine-induced proarrhythmia in the emergency room. AB - The arrhythmogenic hazard of adenosine treatment in an emergency room (ER) has not been established. Thus, in this study, we set out to prospectively determine the prevalence and clinical consequences of the arrhythmogenic effects associated with urgent adenosine treatment in the ER. One hundred and sixty consecutive patients treated with adenosine for regular wide or narrow complex tachyarrhythmias at our ER were included in the study. An initial bolus of 3 mg of adenosine was used, up to a maximum dose of 18 mg (mode 6 mg). Proarrhythmia was defined as the new appearance of any brady- or tachyarrhythmia within 1 minute from the bolus administration of adenosine. Of the 160 study patients, 84% had narrow complex tachycardia and 16% had wide complex tachycardia. Adenosine was effective in the diagnosis and/or treatment of the underlying arrhythmia in 92%. The overall prevalence of adenosine-induced proarrhythmia was 13%, including prolonged AV block inducing asystole > 4 seconds (7%), paroxysmal atrial fibrillation (1%) and non-sustained ventricular tachycardia (5%). All adenosine induced arrhythmias were transient and subsided spontaneously. It is concluded, firstly, that adenosine-induced proarrhythmia proved to be frequent in a consecutive ER series, and included potentially dangerous arrhythmias. Secondly, nevertheless, all adenosine-induced arrhythmias subsided spontaneously and did not require treatment. Therefore, urgent adenosine treatment is safe and can be recommended in an emergency setting, provided a strict protocol of administration under close monitoring by highly trained personnel. PMID- 11436924 TI - Self-reported abstinence effects in the first month after smoking cessation. AB - The present study evaluated self-reported subjective complaints (29 single items and 11 scales) at precessation, on quit day, and on Days 1, 2, 3, 7, 14, 21, and 28 after cessation in 46 healthy quitters who remained abstinent for the first month after cessation (biochemically confirmed). Also tested on the same schedule were 29 nonsmokers matched for age and gender. Specific criteria were set for transient and offset effects based on the direction, magnitude, and time course of changes in symptoms after cessation. Results indicated that single-item anger, anxiety, depression, difficulty concentrating, irritability, restlessness, dizziness, and nausea, and the Shiffman-Jarvik Stimulation/Sedation Subscale, the Perceived Stress scale, and the POMS anger, confusion, and tension subscales met the criteria for transient effects, and that single-item desire to smoke, cough, and headache, and the Shiffman-Jarvik Psychological Subscale met the criteria for offset effects. These findings help to clarify which subjective complaints after smoking cessation are transient effects and which are offset effects, a distinction with important implications for understanding nicotine dependence and for designing pharmacological and nonpharmacological interventions for smoking cessation. PMID- 11436925 TI - Weight locus of control and weight-related attitudes and behaviors in an overweight population. AB - Rotter's [Psychol. Monogr. 80 (1966)] construct of internal-external control of reinforcement led to the conceptualization of locus of control as a personality construct that can be used to predict behavior. More specific measures of locus of control in particular behavioral domains have followed. In the present study, the Weight Locus of Control Scale (WLOC) was used to predict weight-related attitudes and behaviors of overweight individuals, as well as their responses to health education materials (HEM) on weight loss. The WLOC scores predicted responses to baseline weight-related measures such as etiology of obesity, confidence in weight loss behaviors, and behavioral intention. In addition, WLOC scores predicted participants' reactions to the HEM, as well as the actual number of weight loss ideas from the HEM that the participants tried by the 1-month follow-up assessment. The results are discussed in terms of the validity of the WLOC and implications for future development of effective HEM. PMID- 11436926 TI - Characterizing female bipolar alcoholic patients presenting for initial evaluation. AB - This study examined gender differences of age and race-matched group of bipolar disorder (BPO) patients with comorbid alcohol dependence (AD; n = 65; males = 35, females = 30) to a group of BPO patients without comorbid AD (n = 61; males = 22, females = 39). The two groups were also similar on marital status and frequency of BPO subtypes. The results revealed that female bipolar alcoholic patients were more likely to report depressive symptoms as compared to either male bipolar alcoholics or both male and female non-alcoholic bipolar patients. When compared to male bipolar alcoholics, they had higher frequency of depressed mood, slow motor behavior, low self-esteem, decreased libido, decreased appetite, and higher general anxiety symptoms. On the other hand, female bipolar alcoholics differed from female non-alcoholic bipolar patients on reports of mood lability, depressed mood, low self-esteem, suicidal indicators, decreased libido, and general anxiety symptoms. These results raise the question of whether alcohol increases the frequency of depressive symptoms among female bipolar patients. PMID- 11436927 TI - Trends in parent and friend influence during adolescence: the case of adolescent cigarette smoking. AB - A common characterization of adolescence is that parent influence decreases and friend influence increases as adolescents age. From that, we hypothesized that the association between parent and adolescent smoking decreases and the association between friend and adolescent smoking increases as adolescents become older. The hypothesis is tested with data from The National Longitudinal Study of Adolescent Health. Adolescent smoking is measured as progressions to more frequent smoking and as continuations from prior smoking levels. There is no support for the hypothesis, a finding consistent with the earlier panel study that tested it. The age-specific findings are discussed in the context of programs designed to influence adolescent cigarette smoking and why the hypothesis that drove this study was not confirmed. Among supplementary findings reported is that adolescent smoking is more influenced by friend smoking than by parent smoking. PMID- 11436928 TI - The relationship of perceived social support to substance use in offspring of alcoholics. AB - The present study examined the relationship of perceived social support (PSS) to substance use among adolescents with [family history positive (FH+)] and without [family history negative (FH-)] a family history of alcohol dependence. A sample of 144 adolescent offspring of alcoholic fathers and a control group (n = 125) were assessed in relation to substance use and their PSS from both family and friends. The effects of gender and age were also examined. Overall, friend PSS was higher than family PSS, regardless of paternal history. Females scored higher than males on friend PSS. Older adolescents (19-21 years old) perceived higher social support from friends than younger adolescents (14-15 years old). Heavy marijuana and tobacco users reported lower family and friend PSS than light users, while heavy alcohol users reported higher friend PSS than light users. Young FH+ adolescents reported using marijuana at an earlier age than young FH- adolescents. Since the influence of friend PSS on substance use provides a more complex picture than family PSS among adolescents, future analyses will focus on the underlying mechanisms of peer influence on drinking and drug use. PMID- 11436929 TI - Hyperventilation, anxiety sensitivity, and the expectations for alcohol use: subjective and physiological reactivity to alcohol cues. AB - The present study examined the relation between alcohol-related expectancies, body sensation fear and reactions to cues for alcohol following a hyperventilation task. Forty-two undergraduate students participated for course credit. Each student hyperventilated for 5 min, paced at a rate of 30 breaths per minute. Following hyperventilation, each student was exposed to containers with alcohol (beer and wine coolers), with subjective urge to consume and heart rate measures taken. Path analysis supported models associated with tension reduction and self-focused attention expectancies as significant contributors to increased urge to consume alcohol and lowered heart rate following hyperventilation. However, social-anxiety-related expectancies failed to demonstrate a relationship. These results suggest that additional work on the tension reduction model of alcohol use should examine physiological stressors in association with subject characteristics such as proneness to experience panic symptoms. PMID- 11436930 TI - The College Alcohol Problems Scale. AB - A short, reliable two-factor instrument measuring drinking-related negative consequences was developed from a previous measure using two samples of college students. In Study I, data on alcohol use and problems associated with alcohol use were collected on 382 introductory psychology students. The original College Alcohol Problems Scale (CAPS) was tested and found to fit the data poorly. Sequential methods were used to develop a revised instrument. Principal components analyses (PCA) on half of the sample were conducted on 20 items written to measure negative consequences related to college student drinking. Results indicated a two-factor solution measuring social and emotional problems. Confirmatory factor analyses (CFA) on the other half of the sample confirmed the two-factor structure. Further refinement of the instrument resulted in the revised CAPS (CAPS-r), an eight-item two-factor scale. In Study II, the response format was altered to coincide with the Young Adult Problem Screening Test. A total of 726 students completed the instrument as part of a university-wide random sample. CFA showed that the hypothesized model fit well across all measures of model fit and the factor structure was invariant across gender. Additional analyses revealed that the scale was internally consistent and externally valid. A short reliable and valid measure of alcohol-related problems is needed to enable low-cost data collection on college campuses across the nation, as well as to facilitate program evaluation and routine epidemiological surveillance and monitoring. PMID- 11436931 TI - Does a reduced sensitivity to bitter taste increase the risk of becoming nicotine addicted? AB - Cigarette smoking appears to be on the increase in adolescents. The initiation of regular smoking nearly always begins before adulthood. It is therefore crucial to find ways of identifying those children most vulnerable to nicotine addiction and prioritizing them for preventive measures. We hypothesized that individuals who, in a simple taste test, perceive phenylthiocarbamide (PTC) as bitter may find the taste of cigarettes aversively bitter and could therefore have a reduced vulnerability to nicotine addiction compared to nontasters, who would be the group at greater risk of addiction. We studied 242 Plains American Indians, 136 women and 106 men aged 18-59 years, and found that (allowing for gender differences and the possible direct effects of smoking on taste) the proportion of PTC nontasters to tasters in smokers, even light smokers, was significantly greater than in both nonsmokers and social smokers (chi2= 15.875, 4 df; P=.003), suggesting that nontasters, who are not aversive to the bitter taste of cigarettes, may be more at risk for heavy smoking and therefore more vulnerable to nicotine addiction. PMID- 11436932 TI - Retrospective study of the association of stress and smoking during pregnancy in rural women. AB - Women who continue to smoke during pregnancy put themselves and their fetuses at serious risk for complications. Various smoking cessation programs have been designed that specifically target pregnant smokers. Longitudinal studies, however, have shown that there is a group of women who are unable to quit smoking while pregnant. Women from a rural area of the Mid-West (N=299) were interviewed postpartum to determine the stresses these women experienced prenatally and the association of the stress with continuing to smoke during pregnancy. Subjects were divided into three groups: Nonsmokers, Quitters, Smokers. This study not only confirms other reports that these women are more stressed but also documents some of the major stressors. Statistically significant differences were found between groups for financial worries (P=.0002), problems with the family (P<.001), and domestic violence (P<.001). Assessing pregnant women for stress and, especially, domestic violence should be part of the implementation of the Clinical Practice Guidelines for Smoking Cessation. PMID- 11436933 TI - Patterns of adolescent alcohol, cigarette, and marijuana use over a 6-month period. AB - In the current study, alcohol, cigarette, and marijuana use among adolescents (N= 794, 48.6% female) was assessed at two time points, and four patterns of use were identified: (1) abstainers: no lifetime use; (2) new users: no use at baseline but had used by the follow-up period; (3) experimenters: use prior to the baseline but no use in the period up to the follow-up; and (4) consistent users: self-report of use prior to the baseline and the follow-up. Mean levels of psychosocial variables (mastery, self-esteem, and parental social support) were compared across the four patterns of use for each substance. Only analyses including parental social support as the dependent variable were significant. Adolescents with higher levels of social support were more likely to be classified as abstainers or experimenters of alcohol than consistent users. More frequent users of cigarettes at baseline were likely to be classified as frequent users at the follow-up. The discussion focuses on the identification of the situational context of substance use for alcohol, cigarettes, and marijuana. PMID- 11436934 TI - Project EX: outcomes of a teen smoking cessation program. AB - Project EX is an eight-session teen school-based clinic tobacco use cessation program that involves the inclusion of enjoyable, motivating activities ("games," "talk show," and alternative medicine-type) to try to enhance quit rates among youth. This clinic program was tested in a three-group experimental design: clinic-only, clinic plus a school-as-community (SAC) component, and standard care control. Eighteen schools were assigned to the three conditions using a randomized block design. A total of 335 smokers participated in the study, making this the largest controlled teen smoking cessation field trial conducted to date. Seventeen percent of the smokers enrolled in the clinics had reports of having quit smoking for at least the last 30 days at 3-month follow-up (5 months after the program quit day), compared to only 8% of the control condition smokers over than same time period. The Project EX clinic component appears to be an effective means of tobacco use cessation among teens. PMID- 11436935 TI - The relation between intake patterns and substance abuse in a student sample. AB - The purpose of the present study was to identify intake patterns related to substance abuse in a sample of Mexican students. Self-report questionnaires were administered to 897 students in a technical high school. A sample of 300 alcohol and drug users was selected for the present analysis. The sample consisted of 90% alcohol users, 60% male, their mean age being 18.67 (+/-2.25) years old, and mean education being 10.44 (+/-0.67) years. Over 60% met the DSM IV criteria for substance abuse, and almost half of the sample has experienced at least one adverse consequence due to substance use. More than half of the alcohol users drink five or more drinks per drinking occasion. Both substance abusers and males reported more adverse consequences, more frequent, and higher levels of consumption than nonabusers and females, respectively. Results from the logistic regression indicated that four factorial scales derived from the Inventory of Drug-Taking Situations (IDTS) [Annis, A., & Martin, G. (1985). Inventory of Drug Taking Situations. Toronto: Addiction Research Foundation.] were appropriate predictors of substance abuse. The scales Temptation and Boredom, Family Conflict, Good Mood and Testing Control, and Physical Discomfort and Conflict at School were related to substance abuse, correctly classifying 75% of substance abusers. Information about intake situations associated to substance abuse can be useful in the planning of school-based prevention campaigns. PMID- 11436936 TI - Persistent effects of a message counter-marketing light cigarettes: results of a randomized controlled trial. AB - In a randomized, controlled trial, a national sample of smokers of Light cigarettes heard by telephone a "radio message" counter-marketing Light cigarettes. This message caused immediate changes in beliefs. Follow-up telephone interviews were done about 7 months later. The Message Group (N = 181) was more likely than the Control Group (N = 85) to report that (a) one Light equaled one Regular in tar yield to smokers, (b) Lights did not decrease health risks, and (c) they wanted to give up smoking (P<.05); they did not report greater quitting or intention to quit, or greater knowledge of filter ventilation. Systematic counter-marketing of Lights is recommended. A telephone-based exposure and follow up procedure could be a good way to study message effects. PMID- 11436937 TI - Overdosing among young injection drug users in San Francisco. AB - Heroin overdoses increased sharply in the US in the 1990s, but few studies have addressed overdose risk. We examined overdosing and injection-related risk behavior in young injection drug users (IDUs). We interviewed all consenting injectors under age 30 at needle exchanges and youth outreach sites in San Francisco. Their median age was 22, and their median number of years of injecting was 4. About 48% reported at least one overdose, with a median of two overdoses reported. Overdosing was associated with injecting "speedballs" (i.e. mixtures of heroin and cocaine), with borrowing syringes, and (with P-values of borderline statistical significance) with heroin injection and with gay or bisexual behavior. It was not associated with age, sex, years of injecting, or frequency of injecting. In multivariate analysis, only borrowing syringes and gay or bisexual behavior were independent statistically significant predictors, probably because gay and bisexual subjects were more likely to be heroin or "speedball" injectors. Most subjects (65%) reported that they had not received medical attention at time of last overdose. Risk of overdose in young injectors is acute and closely associated with HIV risk. HIV interventions should include overdose prevention. Emergency response protocols should minimize risk of arrest. Injectors and providers should be trained in overdose prevention, and developing overdose interventions should be a priority among drug educators. PMID- 11436938 TI - The relationship between self-reported cocaine withdrawal symptoms and history of depression. AB - This study examined the relationship between cocaine withdrawal and lifetime history of depression (major depression, dysthymia). Participants with a history of regular cocaine use (n = 146) were administered the Structured Clinical Interview for the DSM-IV (SCID) and were asked to recall whether they experienced any of the six DSM-IV cocaine withdrawal symptoms. Results of bivariate analyses demonstrated that those meeting criteria for the cocaine withdrawal syndrome (dysphoria plus two or more other symptoms), in comparison to those who did not, were significantly (P<.001) more likely to have a lifetime history of depression. Lifetime history of depression was also more common in those individuals reporting the withdrawal symptoms of "dysphoria" (P<.001), "insomnia/hypersomnia" (P<.05), "vivid unpleasant dreams" (P<.01), and "psychomotor agitation/retardation" (P<.01). These relationships remained significant after controlling for demographics, severity of addiction, and the presence of opiate, alcohol and cannabis dependence or abuse. The withdrawal symptoms of "fatigue" and "increased appetite" were not associated with mood history. Results suggest that lifetime history of depression is strongly related to whether or not a cocaine abuser self-reports withdrawal symptoms. Several competing hypotheses regarding the nature of this relationship are discussed. PMID- 11436939 TI - Clinical and psychosocial characteristics of substance-dependent pregnant women with and without PTSD. AB - The present study compared psychiatric and psychosocial functioning in 123 pregnant opiate- and/or cocaine-dependent women with and without a comorbid diagnosis of posttraumatic stress disorder (PTSD). Participants were enrolled in a comprehensive perinatal drug treatment program and completed assessments upon admission. Lifetime diagnostic prevalence of PTSD [Structured Clinical Interview for DSM-IV Disorders (SCID) confirmed] among the sample was 19%. Participants with PTSD (n=24) reported greater need for psychiatric treatment, were more likely to report a previous suicide attempt, and had more previous drug treatments than participants without PTSD (n=99). Women with PTSD were twice as likely to have lifetime Axis I and Axis II disorders and had higher rates of abuse than women without PTSD. Lifetime sexual abuse and ASI family/social composite scores were significant predictors of PTSD. Findings suggest that pregnant drug-dependent women with comorbid PTSD may benefit from specialized treatment services for trauma and/or abuse issues. PMID- 11436940 TI - Effect of brimonidine tartrate on ocular hemodynamics in healthy volunteers. AB - While alpha2-adrenergic agonists, such as brimonidine tartrate, significantly reduce the intraocular pressure (IOP), the presence of vasoconstrictor postsynaptic alpha2 receptors on vascular smooth muscle raise the possibility that brimonidine could potentially compromise ocular blood flow. Consequently, the ocular hemodynamic effects of brimonidine were studied in normal subjects. Twelve healthy volunteers were included in this prospective, double-masked, placebo controlled, crossover-designed clinical trial. They received either brimonidine tartrate 0.2% or placebo b.i.d. for 2 weeks. Goldmann tonometry and color Doppler imaging (CDI) were performed at baseline, at 2 hr, 1 week, and 2 weeks after the treatment. Fundus angiography using a scanning laser ophthalmoscope was performed at baseline and 2 weeks after treatment to determine retinal arteriovenous passage time. Brimonidine lowered IOP at 2 hr, 1 week, and 2 weeks (p = 0.058, p = 0.031, and p = 0.022, respectively). Brimonidine did not affect the retrobulbar arterial velocities measured by CDI, nor retinal arteriovenous passage time. In conclusion, two-week treatment with brimonidine reduces IOP and does not reduce the bulk retinal or retrobulbar arterial perfusion in young healthy volunteers. PMID- 11436941 TI - Plasma concentration of topically applied betaxolol in elderly glaucoma patients. AB - Our aim was to study the concentration of betaxolol in plasma after its topical ocular use during the normal 12 hr dosing interval. Twenty microliters of betaxolol 0.5% solution were applied into both eyes of nine glaucoma patients, and the plasma concentrations of the drug were measured 12 hr thereafter using a radioreceptor assay. The same amount of betaxolol was then applied ocularly, and its concentration in plasma was measured at 5, 10, 15, 30 min and 1, 2, 4 and 8 hr thereafter. The mean (SD) concentration of betaxolol in plasma twelve hr after the first dose was 0.4 (0.2) ng/ml. After the second dose, the patients showed a biphasic concentration vs. time curve, the first peak occurring at 8 (4) min, and the second peak at 210 (132) min; the mean (SD) peak concentrations being 1.1 (0.3) and 2.0 (1.1) ng/ml, respectively. The area under the concentration vs. time curve showed a 4-fold variation among our patients. Topically applied betaxolol was rapidly absorbed into systemic circulation, and concentrations were detectable even at 12 hr. The interindividual variation in the systemic absorption of betaxolol was large. PMID- 11436942 TI - Effects of 2% ibopamine on pupil, refraction, anterior segment anatomy and intraocular pressure. AB - The aim of the study was to determine the effects of a dopaminergic drug, 2% ibopamine, on the pupil, intraocular pressure and other ocular and ultrasound biometric variables. Thirty healthy subjects and 15 patients with primary open angle glaucoma, aged from 40 to 78 years (mean age: 59.2 +/- 11), were included in two prospective open controlled trials. In the first, the mydriatic effect of 2% ibopamine and its inhibition and reversibility were evaluated in 15 healthy subjects using the alpha1-adrenergic drug, 0.5% dapiprazole. In the second, refraction, visual acuity, pupil diameter, intraocular pressure and 5 A-scan ultrasound biometric variables were evaluated in 15 healthy subjects and in 15 glaucoma patients. As early as forty min after administration of 2% ibopamine, a marked mydriatic effect (7.3 vs 3.9 mm; P < 0.0001), which was completely inhibited or reversed by 0.5% dapiprazole, was detected. The drug induced no changes in refraction, visual acuity or A-scan ultrasound biometric variables in any of the subjects examined. In healthy subjects, the intraocular pressure values were not changed to a statistically significant extent (13.8 vs 14.8 mm Hg; P = 0.668), whereas a slight, though significant, hypertensive effect (24 vs 22.2 mm Hg; P = 0.002) was observed in the glaucoma patients. The study confirms the intense mydriatic effect of 2% ibopamine with no changes in refraction, visual acuity or A-scan ultrasound biometric variables. The drug has no effect on intraocular pressure in healthy subjects, but induces a significant hypertensive effect in patients with initial glaucoma. This characteristic could be used for early diagnosis of primary open-angle glaucoma. PMID- 11436943 TI - Intraocular pressure and vascular effects of sodium azide in bovine perfused eye. AB - The effects of the nitrovasodilator, sodium azide, on intraocular pressure (IOP) and ciliary vascular tone were compared. IOP was measured in the bovine isolated eye that was perfused via the ciliary artery. Separately, vasodilator effects were assessed after raising the vascular tone using noradrenaline (10 microM). Aqueous humor formation (AHF) rate was estimated by a fluorescein dilution method. Cyclic GMP in the ciliary processes was measured by radioimmunoassay. When compared with controls, sodium azide (10 nmole bolus dose) was found to lower IOP (2.2 +/- 0.3 mm Hg; P < 0.01) via a reduction in AHF (12.19 +/- 0.26 microl/min to 6.36 +/- 0.53 microl/min; P < 0.001). Azide (1 micromole) also reduced ciliary vascular resistance (81.0 +/- 5.5%; P < 0.01). However, the drug was 20x more potent as an ocular hypotensive than as a vasodilator (ED50 0.28 nmole on IOP, 5.55 nmole on vascular effect). Azide (10 nmole) also increased levels of ciliary cyclic GMP (127 +/- 17 fmol/mg protein to 233 +/- 27 fmol/mg protein; P < 0.01). The IOP-lowering effect of azide does not appear to depend on its ability to activate guanylyl cyclase (GC) in vascular smooth muscle, but rather is likely a consequence of direct activation of ciliary epithelial GC. PMID- 11436944 TI - Assessment of systemic adverse reactions induced by ophthalmic beta-adrenergic receptor antagonists. AB - To assess quantitatively the risks of ophthalmic beta-blocking agents for cardiovascular and respiratory adverse reactions, we analyzed the binding kinetics of beta-blocking agents to the beta-1 and beta-2 adrenoceptors. The relationship between the occupancies for beta-1 and beta-2 adrenoceptors and the effects on the exercise pulse rate or the forced expiratory volume in one second (FEV1) after topical administration of carteolol, befunolol, timolol and betaxolol was analyzed using a ternary complex model. The beta-1 and beta-2 receptor occupancies after ophthalmic administration were calculated to be quite high as well as those after oral administration. The maximum occupancies for beta 1 and beta-2 receptors after ordinary ophthalmic administration were 52% and 88% for carteolol, 52% and 61% for befunolol, 62% and 82% for timolol, and 44% and 3% for betaxolol, respectively. Concave relationships were obtained between a decrease in exercise pulse rate and the beta-1 receptor occupancy and between a decrease in FEV1 and beta-2 receptor occupancy, respectively. Nasolacrimal occlusion was estimated to decrease the exercise pulse rate and FEV1 by 65% and 50%, respectively. The beta-1 and beta-2 adrenoceptor occupancies were proved to be the most appropriate indicators for cardiac and pulmonary adverse reactions evoked by ophthalmic beta-blocking agents. PMID- 11436945 TI - Maintenance of mydriasis with one bolus of epinephrine injection during phacoemulsification. AB - Surgically-induced miosis commonly occurs during cataract extraction surgery, complicating removal of lens cortex and placement of a posterior chamber intraocular lens. To maintain intraoperative mydriasis, one bolus of epinephrine injection was used in our study. The pupillary response to various doses of intracameral epinephrine (0.1 ml of 1:25,000, 1:50,000, 1:100,000, 1:200,000, 1:400,000) was assessed in 60 consecutive patients. The pupil size was measured just prior to the incision, one min after epinephrine injection, after phacoemulsification and after irrigation/aspiration. There was no significant difference among the mean mydriatic responses to the epinephrine concentrations we tested. The 1:400,000 concentration appeared to be as effective as 1:25,000, but two cases of the 1:400,000 group failed to maintain the pupil diameter after irrigation/aspiration. In addition, we found that blood pressure did not elevate after injection of any concentration of epinephrine. We concluded that one bolus of an extremely dilute concentration of epinephrine (i.e., 1:400,000) injection might be effective in maintaining mydriasis during cataract surgery without systemic side effects. PMID- 11436946 TI - Differential effects of alpha-adrenoceptor agonists on human retinal microvessel diameter. AB - The effects of locally administered brimonidine, clonidine, and p-aminoclonidine on microvessel caliber were compared in human retinal tissues grafted into the hamster cheek pouch. Clonidine and p-aminoclonidine, but not brimonidine, potently constricted human retinal microvessels over a broad concentration range. All three agonists elicited significant vasoconstriction in naive hamster cheek pouch microvasculature. The alpha2-adrenoceptor antagonist, rauwolscine, inhibited p-aminoclonidine-induced constriction in naive hamster cheek pouch microvessels, but not p-aminoclonidine-induced effects in retinal grafts. Selective alpha1-adrenoceptor agonists evoked vasoconstriction in retinal grafts only at relatively high concentrations. These differential effects on the retinal microvasculature could not be readily explained solely on the basis of alpha1- or alpha2-adrenoceptor involvement. Clonidine, p-aminoclonidine and brimonidine are also imidazoline derivatives that interact with putative non-adrenergic imidazoline-sensitive binding sites, the so-called I1-imidazoline binding site subtype implicated by some investigators in mediation of peripheral vasoconstriction. As with p-aminoclonidine, the potent vasoconstriction in human retinal microvasculature elicited by moxonidine, an alpha-adrenergic agonist that has also been reported to exhibit selectivity for putative I1-imidazoline binding sites, was not inhibited by the selective alpha-adrenoceptor antagonist, rauwolscine, nor by idazoxan, an antagonist characterized as having substantial activity at putative I2-imidazoline binding sites. These data suggest the possible involvement of an unconventional non-adrenergic imidazoline-sensitive pathway in regulation of microvascular responses in the inner retina, and that drug activity mediated via such an imidazoline-sensitive component could potentially evoke deleterious effects in the retinal microvasculature. PMID- 11436947 TI - Peroxiredoxin in bovine ocular tissues: immunohistochemical localization and in situ hybridization. AB - Peroxiredoxins are widely distributed in nature and constitute a molecular family of antioxidant enzymes which decompose hydrogen peroxide and alkyl hydroperoxides. We have previously characterized a peroxiredoxin from bovine ciliary body and deduced its amino acid sequence from analysis of cDNA clones encoding the protein. In this work, we investigated the immunolocalization of this novel antioxidant enzyme and its mRNA expression in bovine eye tissues. High levels of immunoreactivity and mRNA for the enzyme were detected in corneal epithelium. Distinct immunoreactivity and mRNA expression for peroxiredoxin were also detected in uveal tissues, some of the retinal cell layers and ocular vasculature. PMID- 11436948 TI - Treatment of ocular toxocariasis with albendazole. AB - The purpose of this study was to evaluate the efficacy of combined albendazole and steroid treatment for uveitis caused by Toxocara canis in immunocompetent patients. Five patients (7 eyes) with ocular larva migrans syndrome (OLM) were used in this study. Toxocariasis was suspected based on clinical manifestations and confirmed by anti-toxocara IgG and Western blot analysis. Systemic albendazole (adults: 800 mg b.i.d.; children: 400 mg b.i.d.) was given in conjunction with steroids. Visual acuity before and after therapy, inflammatory response, side effects and toxicity were evaluated. Treatment resulted in an improved visual acuity in all patients. Mean initial Snellen visual acuity was 20/40, and mean final acuity was 20/20. There were no recurrences of uveitis throughout the observation period (average: 13.8 months; range: 3 days to 24 months). These findings suggest that albendazole, in combination with systemic steroids, is a useful regimen to treat ocular larva migrans syndrome. PMID- 11436949 TI - Evaluation of intraocular pharmacokinetics and toxicity of prinomastat (AG3340) in the rabbit. AB - To determine the ocular pharmacokinetics, physiological and histological effects of prinomastat (a matrix metalloprotease inhibitor), a total of seventy-seven eyes of New Zealand White rabbits received intravitreous and subtenon injections of prinomastat or of acidified water vehicle as control, Doses of 0.5 mg in 0.05 mL of prinomastat or acidified water were used for intravitreal injection. For the subtenon injections, doses of 5 mg prinomastat in 0.5 mL of acidified water were administered in the superotemporal quadrant. Intraocular pharmacokinetics were determined by analyzing vitreous samples at different postinjection time points using Liquid Chromatography-Mass Spectroscopy/Mass Spectroscopy (LC MS/MS). The toxicity was evaluated by biomicroscopy, electroretinography (ERG), pneumatonometry, and histology. No toxicity was found with either administration method. At day 14 after intravitreal injection, levels of prinomastat in the vitreous and choroid were 1.4 ng/mg and 7.8 ng/mg, respectively. The retinal levels of prinomastat were 22 ng/mg at 24 hr and dropped below 1 ng/mg at 48 hr. Prinomastat remained well above minimum effective concentration in the choroid for at least four weeks after a single intravitreal injection, suggesting that local intravitreal injection may have potential in treating choroidal neovascularization. PMID- 11436950 TI - Psychopharmacology news. PMID- 11436951 TI - Update and recommendations for the use of antipsychotics in early-onset psychoses. AB - A review was undertaken of studies evaluating the efficacy and tolerability of antipsychotic medications for the management of psychosis in children and adolescents. All identified published and unpublished studies from 1996 onward were included for review. The search located one randomized control trial, seven open-label trials, six retrospective chart reviews, and nine case reports. The studies assessed the use of haloperidol, clozapine, risperidone, olanzapine, and quetiapine in the management of psychosis in children and adolescents. Most studies reported reasonable treatment response; however, extrapyramidal side effects, sedation, and weight gain are concerning. This points to the need for appropriate baseline assessments prior to initiating treatment with these agents. Particular attention should be given to assessment of the extrapyramidal system as well as to baseline weight, lipid profile, and blood glucose. Further study is needed to refine the use of antipsychotic medications in children and adolescents in order to minimize adverse effects while conferring an optimum therapeutic response. The importance of instituting effective early treatment in youth with psychoses is an important goal that may serve to lessen the long-term morbidity of the illness. PMID- 11436952 TI - Sertraline effects in adolescent major depression and dysthymia: a six-month open trial. AB - This 6-month open-label study evaluated the efficacy, tolerability, and safety of sertraline in 21 adolescent psychiatric outpatients, ages 12 to 18 years, diagnosed with major depressive disorder (MDD, n = 13) or dysthymic disorder (DD, n = 8). Both groups showed clinically significant improvements on the Hamilton Depression Scale (HAM-D), Hamilton Anxiety Scale, and the Clinical Global Impression Scale-Severity (CGI-S). The MDD group showed maximal clinical response (based on the method of last observation carried forward) on the HAM-D and CGI at weeks 12 (76.9%) and 20 (76.9%), respectively. Response rates were maintained at week 24 with all six MDD study completers (100%) responding to treatment. The DD group achieved maximal response on the HAM-D (100%) and the CGI (75%) at week 6. Response rates in this group did not remain as elevated over time with two out of three (66.7%) DD study completers responding to treatment at week 24. Generally, sertraline was safe and well tolerated. Most adverse events were mild to moderate in severity and resolved with no action taken. Results suggest that sertraline may be efficacious in acute and continuation treatment of MDD in adolescents. DD patients showed evidence of clinical response and improvement, particularly in the acute treatment phase. Incorporating a longer evaluation period in the study of antidepressant therapy for adolescents with MDD and/or DD is emphasized. PMID- 11436953 TI - Sertraline in the treatment of restricting anorexia nervosa: an open controlled trial. AB - Few studies to date have examined the usefulness of selective serotonin reuptake inhibitors (SSRIs) in an outpatient multidisciplinary treatment of anoxrexia nervosa. The present study consists of an open, controlled trial of sertraline in a sample of 11 restricting-type anorexics, treated in an outpatient setting, compared with a control group of patients with similar characteristics. All subjects were assessed by means of a structured interview and self-reported questionnaires at baseline and after 14 weeks of treatment. Body weight and diagnostic status were evaluated again after a 64-week follow-up. At the 14-week follow-up, the sertraline group reported a significantly greater improvement of depressive symptoms, ineffectiveness, lack of interoceptive awareness, and perfectionism when compared to the control group. Both groups reported a significant improvement in body weight. At the 64-week follow-up, only one patient of the sertraline group and five patients of the control group still had a full diagnosis of an eating disorder. Further studies are necessary to define the role of sertraline and of other SSRIs in the treatment of anorexia nervosa. PMID- 11436954 TI - Paroxetine in child and adolescent outpatients with panic disorder. AB - Paroxetine has repeatedly been shown to be effective in the treatment of panic disorder (PD) in adults, and, according to previous case observations, it may be useful in treating children and adolescents with PD as well. This preliminary naturalistic study examines effectiveness and safety of paroxetine in the treatment of children and adolescents with PD. A chart review was conducted on 18 patients with Diagnostic and Statistical Manual of Mental Disorders PD admitted to the Division of Child Neurology and Psychiatry and to the Department of Psychiatry at the University of Pisa. Paroxetine was given at an initial mean dosage of 8.9 +/- 2.1 mg/day and was gradually increased up to 40 mg/day, depending on clinical response and side effects. Clinical status was assessed with the Clinical Global Impression (CGI) and adverse effects were assessed retrospectively at each visit. Patients with final CGI-Improvement scores of 1 or 2 were considered responders. Mean paroxetine treatment duration was 11.7 +/- 8.3 months, with a mean final dosage of 23.9 +/- 9.8 mg/day (range, 10-40 mg/day). No patient had to interrupt the treatment because of side effects. Fifteen patients (83.3%) were considered responders. The mean change on the CGI-Severity scale was statistically significant (p < 0.0001). Paroxetine was well tolerated and effective in the treatment of PD in these children and adolescents. PMID- 11436955 TI - A retrospective study of citalopram in adolescents with depression. AB - Recent evidence suggests that the selective serotonin reuptake inhibitors are safe and efficacious in treating juveniles with depression. However, citalopram has not been reported in adolescents with depression. This study assessed the effectiveness and tolerability of citalopram in all adolescents with depressive disorders treated naturalistically in a community mental health center during a 1 year interval. Medical charts were retrospectively reviewed for 21 adolescents treated with citalopram for major depression (n = 14), bipolar depression (n = 4), or dysthymia (n = 3). An independent rater compared last visit to baseline depression using the Clinical Global Impression (CGI) Severity and Improvement scales. Adolescents received citalopram for an average of 128.5 +/- 84 days at a final average dose of 26.5 +/- 13.1 mg/day. Sixteen of these 21 adolescents (76%) exhibited much to very much improvement as measured by the CGI, and severity of depression diminished significantly (z = 3.007, p < 0.0026). Mild side effects, including headaches, dizziness, nausea, sedation, agitation, and sweating were reported by 7 (33%) of the patients. These data suggest that citalopram may be effective, safe, and well tolerated in the treatment of adolescents with depressive disorders and that controlled trials are warranted in this population. PMID- 11436956 TI - An open-label trial of tomoxetine in pediatric attention deficit hyperactivity disorder. AB - OBJECTIVE: To collect pilot data assessing the safety, tolerability, and efficacy of tomoxetine, a nonstimulant norepinephrine enhancer, in pediatric attention deficit hyperactivity disorder (ADHD). METHODS: An open-label trial of tomoxetine in pediatric ADHD was conducted as part of a multisite clinical trial. Following a baseline assessment, an ascending dose titration was completed during 10 weekly visits. RESULTS: Ten subjects were enrolled at baseline, with eight completing the study. Seven of the eight remaining subjects met efficacy criteria. Significant decreases in symptom severity ratings by parents and study investigators were found. The medication was well tolerated, with transient appetite suppression the most frequently reported side effect. However, subjects' weights remained stable across study visits. DISCUSSION: These preliminary findings suggest that tomoxetine may hold promise as a treatment for pediatric ADHD. PMID- 11436957 TI - Efficacy of Adderall and methylphenidate in attention deficit hyperactivity disorder: a reanalysis using drug-placebo and drug-drug response curve methodology. AB - Because methylphenidate (MPH) is currently the most widely prescribed medication for attention deficit hyperactivity disorder (ADHD), several studies have used this as the touch-stone for evaluating the efficacy of a newer stimulant, Adderall. In a parallel-groups study of MPH (n = 20), Adderall (n = 20), and placebo (n = 18), Pliszka et al. (2000) reported that both medications were superior to placebo in improving parent, teacher, and clinician ratings of ADHD and associated behaviors. Compared with MPH, Adderall led to significantly more improvements in teacher and clinician ratings. The present study extends these results by addressing the issue of clinical significance using drug-placebo and drug-drug response curve analyses of the same data. The goal of this method is to answer the following questions about drug-placebo or drug-drug differences: Is the effect clinically meaningful? What does the effect tell us about individual responses? Is the effect due to symptom improvement, the prevention of worsening, or both? Our results show that the efficacy of Adderall to improve functioning is seen throughout the full range of improvement scores. In contrast, MPH showed a substantial effect for "mildly" and "much improved" but not for "very much improved." Our analyses also show that both Adderall and MPH prevent worsening of symptoms. They further suggest that, compared with the Conners Teacher Rating Scale, the Clinical Global Impressions scale may be more sensitive to improvements at the "well end" of the spectrum of functioning. PMID- 11436958 TI - Amotivational syndrome associated with selective serotonin reuptake inhibitors in children and adolescents. AB - A frontal lobe syndrome has previously been reported in adults treated with selective serotonin reuptake inhibitors (SSRIs), but not in children. Five typical cases of apathy and lack of motivation, one accompanied by disinhibition, are described in a child and four adolescents. Symptoms were dose related and reversible. The subtlety of symptoms, lack of insight in patients, disabling effects, and delayed onset indicate a need for clinicians to inform families of these potential symptoms when SSRIs are prescribed. PMID- 11436959 TI - Quetiapine treatment of children with Tourette's syndrome: report of two cases. AB - Two children with Tourette's syndrome and comorbid disorders were treated with quetiapine, an atypical antipsychotic successfully used in patients with psychoses and schizophrenia with low incidence of extrapyramidal side effects. Clinical observations and standardized rating scales suggested that this drug produced beneficial effects on tics and other symptoms. Adverse effects (at low doses) were minimal. Because it was suggested that tic efficacy of the newer antipsychotics was related to higher D2 occupancy (with the exception of quetiapine and clozapine, which have relatively low D2 activity), it is hypothesized that tic patients are D2 sensitive and need lower doses of medications. These children were treated naturalistically and were reported retrospectively because of their encouraging outcomes. However, these findings should be interpreted with caution, because no contrast groups, drug withdrawal, or placebo were utilized. Controlled studies are needed to determine the efficacy of quetiapine in the treatment of Tourette's syndrome. PMID- 11436960 TI - Pharmacokinetic interactions between cyclosporine and bupropion or methylphenidate. AB - A 10-year-old boy with a history of heart transplantation had a potentially life threatening decrease in his cyclosporine (CSA) blood levels during administration of bupropion. Subsequently he had an increase in CSA levels while receiving methylphenidate. These occurrences represent potential drug-drug interactions in children not previously documented in the literature. The CSA-bupropion and CSA methylphenidate interactions merit further investigation, particularly because psychotropic agents are often prescribed in combination with immunosuppressants in transplantation patients of all ages. PMID- 11436961 TI - Myoclonus during prolonged treatment with sertraline in an adolescent patient. AB - A 15-year-old female adolescent with depression developed myoclonus after uninterrupted treatment with sertraline over 6 years. She was also receiving methylphenidate. Withdrawal of sertraline and continuation of methylphenidate did not result in any improvement. Treatment with valproic acid resulted in improvement of the movement disorder. This report suggests that myoclonus may be a side effect of sertraline in some adolescents. Further, we hypothesized that extended treatment over several years, young age, and a compromised central nervous system due to underlying disorders may be risk factors for the development of this side effect. PMID- 11436962 TI - Reboxetine as a potentially effective treatment for attention deficit hyperactivity disorder. PMID- 11436963 TI - Effectiveness of quetiapine in the management of psychotic depression in an adolescent boy with bipolar disorder, mixed, with psychosis. PMID- 11436964 TI - Quetiapine resolves psychotic depression in an adolescent boy. PMID- 11436965 TI - Are autoantibodies present in patients with subacute and chronic urticaria? AB - Since several forms of autoimmunity have been associated with urticaria, we performed a detailed survey of autoantibodies in patients with idiopathic subacute and chronic urticaria. Sera from 25 consecutive patients referred for evaluation of urticaria were tested for the presence of autoantibodies and compared to sera from seventy-five control samples examined from individuals being treated for other allergic diseases. Study patients ranged in age from 15 to 73 years, with a mean of 48. One patient had a diagnosis of inflammatory bowel disease and one had multiple myeloma, but otherwise there were no other diagnoses of disease specifically involving immunity other than atopy. No study patients had a concurrent diagnosis of autoimmune thyroid disease. The test sera were examined for autoantibodies and for antibodies to H. pylori. Antibodies to thyroid peroxidase (TPO) were found significantly (p < 0.01) more common in urticaria (20%] than in controls (0%). Rheumatoid factor(RF) was also found in significantly (p < 0.05) increased in urticaria (16%) compared to controls [0%). Neither H. pylori antibody nor other autoantibodies were present in significant numbers of urticaria patients when compared to controls. Tested autoantibodies included those to thyroglobulin, sDNA, SSA/SSB, ENA, cardiolipin, beta2 glycoprotein I, myeloperoxidase, proteinase-3, smooth muscle, ANA, human lysosomal-associated membrane protein, and bactericidal permeability increasing protein. Thus, patients with urticaria were somewhat more likely to have a thyroid autoantibody to TPO or to have RF. This survey demonstrates that while some markers of autoimmunity may be increased in urticaria patients, broad nonspecific autoimmunity is not found. PMID- 11436966 TI - Immunogenicity of hydrolysate formulas in children (Part 2): 41 case reports. AB - Hydrolysate formulas (HF) have been developed with the purpose of reducing the allergenicity of cow's milk proteins, thus providing a suitable formula for feeding babies with cow's milk (CM) allergy (CMA). More recently, used for the treatment of CMA HFs have provoked 211 reactions in babies at high risk of atopy . In this paper we report 81 babies, who attended the Allergy and Clinical Immunology Division of the Pediatric Department of Rome University "La Sapienza" (study children) between June 1997 and October 1998, 41 of whom were tested with Nidina HA (a whey partial HF) and 40 with whole CM. Ninety-seven healthy children with no history of atopy of comparable age and sex formed the control group. All study children reacted to Nidina HA and to CM with similar symptoms. All the control babies tested negative. With these 41 case reports the number of reactions to HFs totals 252. These data strongly indicate that partial HFs may be allergenic not only in an already sensitized individual, but also immunogenic in a predisposed baby. PMID- 11436967 TI - The safety of cluster immunotherapy with aluminium hydroxide-adsorbed honey bee venom extract. AB - Venom immunotherapy (VIT) with aqueous extracts is considered a life-saving treatment in insect allergy, but systemic side effects are quite common, especially in patients sensitized to Apis mellifera venom. Only a few studies are available regarding depot VIT administered according to the standard weekly schedule, while no data are available for cluster VIT with depot preparations. This study was designed to compare the tolerance of aqueous cluster or rush VIT versus cluster depot VIT in patients sensitized to Apis mellifera. Fifty-five patients with a history of systemic reaction (grade II to IV according to Muller) after an Apis mellifera sting were divided at random into 3 groups. Forty patients were treated with aqueous VIT (20 according to a cluster schedule and 20 according to a rush schedule) and 15 with Depot VIT according to a cluster schedule. All local and systemic side effects were recorded. Four patients belonging to the aqueous-cluster, 4 belonging to the aqueous-rush, and 1 belonging to the depot-cluster schedule had large local reactions after VIT (p < 0.009). Systemic side effects were recorded in 1, 7 and 0 patients (p < 0.003) respectively. No statistically significant difference was found between the two cluster regimens in the frequency of side effects. Depot VIT administered according to a cluster schedule was well tolerated, and its tolerance is better than aqueous-rush VIT for Apis mellifera. PMID- 11436968 TI - Hypersensitivity pneumonitis: a comprehensive review. AB - Hypersensitivity pneumonitis is a dynamic and heterogeneous group of diseases resulting from inhalational exposure to a variety of organic and inorganic dusts. In the United States, it has predominantly been considered an occupational disease, such as in farmer's lung, bagassosis, and in the plastics and paint industry with isocyanate exposure. In Japan, however, the most common exposure to antigen has been associated with the home environment. All susceptible patient populations are at potential risk to develop this disease under appropriate antigen exposure. The clinical course can be acute, subacute, or chronic depending on the nature of the antigen and the circumstances under which exposure occurs. If unrecognized, any stage of disease has the potential to progress to endstage pulmonary fibrosis. Proper antigen identification and avoidance requires that the physician be keenly aware of its existence and the importance of the detailed occupational and home environmental history. Without a high degree of suspicion, this disease can be easily overlooked and misdiagnosed as another type of inflammatory lung disease. Although the immunologic mechanisms mediating this illness are complex and poorly understood, strict avoidance of the provocative antigen is required for optimal long-term outcome. PMID- 11436969 TI - Treatment of severe seasonal rhinoconjunctivitis by a combination of azelastine nasal spray and eye drops: a double-blind, double-placebo study. AB - BACKGROUND: Evaluation of combined azelastine nasal spray and eye drops treatment in patients with severe rhinoconjunctivitis. METHODS: Phase III, multicenter, randomized, double-blind study of patients with a history of grass pollen allergy, confirmed by skin testing/specific IgE, total symptom scores > or =6 (ocular) or > or =8 (nasal). Intent-to-treat analysis. RESULTS: 99 patients (azelastine = 53, placebo = 46) enrolled homogeneously from May to September 1997 in 7 venues in France. The efficacy of azelastine was significantly higher compared to placebo (49% vs. 28%, p = 0.04), considering response as a decrease of the total sum of ocular and nasal scores by at least 50% and no use of cetirizine by day 7. The decrease of total ocular and nasal scores by at least 50% at day 7, with cetirizine rescue <3 tablets was higher, but not significantly, in azelastine patients (43% vs. 30%). Cetirizine rescue was more frequent, from day 0 to 7, in the placebo patients (4.9 +/- 5.0 vs. 2.7 +/- 4.1, p = 0.02). Global efficacy was rated higher for azelastine by investigators (26% vs. 10%, p = 0.05) and patients (28% vs. 7%, p = 0.01). Adverse events were burning sensation, "red eyes," nasal irritation, bitter taste. No serious adverse events were reported. Tolerance of azelastine was "very good/good"/"satisfactory" in the majority (62%/82% assessed by investigators, or 55%/79% by patients, respectively). CONCLUSIONS: Combining azelastine eye drops and nasal spray is a safe and effective treatment of severe seasonal rhinoconjunctivitis. PMID- 11436970 TI - Controlled study of preseasonal immunotherapy with grass pollen extract in tablets: effect on bronchial hyperreactivity. AB - BACKGROUND: Based on experimental results, the sublingual route for immunotherapy (IT) has been accepted as a viable alternative to the injection route, but few data on the effects on asthma are so far available. OBJECTIVE: In the present open controlled trial we evaluated whether a preseasonal IT with grass polllen in orosoluble tablets added to pharmacotherapy, can improve non-specific bronchial hyperreactivity. The clinical efficacy was evaluated as well. METHODS: Fifty-one patients (mean age 27.4 years) suffering from rhinoconjunctivitis and/or mild intermittent/mild-persistent asthma due to grass pollen were allocated to two groups receiving pharmacotherapy alone (n = 25) or pharmacotherapy plus IT in tablets (n = 26). A methacholine test was performed in asthmatic subjects out of the pollen seasons at baseline and after 3 years of treatment. Symptom scores and drug intake were evaluated during pollen seasons by a diary card. RESULTS: A significant increase p = .01) in the PD20 at the methacholine test was observed in the IT group compared to the control group. A significant clinical improvement both for rhinitis (p = .001) and asthma (p = .001) was observed in the IT group, and this improvement was paralleled by a clear-cut reduction of drug intake (p = .001). An improvement of rhinitis symptoms without modification of drug intake was seen in the control group (p = .01) The treatment was well tolerated and no relevant side effect was reported during the 3 years. CONCLUSION: The investigated local IT reduced the nonspecific bronchial hyperreactivity. Furthermore, it was clinically effective and safe. PMID- 11436971 TI - Serum soluble tumor necrosis factor receptor levels in patients with seasonal allergic rhinitis. AB - It is known that several inflammatory cells and cytokines play a role in allergic inflammation. Furthermore, there are seasonal changes in several mediators responsible for allergic inflammation. The aim of this study was to determine seasonal changes in serum concentrations of soluble tumor necrosis factor receptors (sTNFRs) and the relationship with disease activity and other inflammation markers such as eosinophil cationic protein (ECP) in patients with seasonal allergic rhinitis. Serum levels of sTNFRI and sTNFRII were measured before and during the pollen season in 18 patients with seasonal allergic rhinitis and in 17 healthy controls by using commercially available enzyme-linked immunosorbent assay (ELISA) kits. The serum levels of sTNFRI, sTNFRII, IgE, and ECP were significantly higher in patients than those in controls before and during season. sTNFRI, sTNFRII, and IgE levels were higher before season, whereas ECP levels were higher during season. We suggest that sTNFRs might play regulatory roles even in early stages of allergic rhinitis when patients do not have clinical symptoms yet. PMID- 11436972 TI - Freezing protects against allergy to Anisakis simplex. AB - BACKGROUND: Allergic reactions to Anisakis simplex have been described following ingestion of fish and were thought to be possible even if seafood is frozen or well-cooked. OBJECTIVE: This study aimed to confirm, by challenge test, that dead A. simplex larvae are not able to trigger allergic reactions in patients with proven hypersensitivity to this parasite. METHODS: Simple-blind, placebo controlled, oral challenge tests with frozen A. simplex larvae were performed in 12 patients who had suffered severe anaphylactic reactions after ingestion of seafood and diagnosed of A. simplex hypersensitivity by skin prick test and specific IgE. If no reaction appeared, they were told to eat frozen seafood. 63 patients who had suffered urticaria or urticaria/angioedema by demonstratred hypersensitivity to A. simplex were also advised to eat frozen seafood. All of them were reevaluated 6 months later. RESULTS: All patients tolerated the dead larvae challenge test. After eating previously frozen seafood at least two times per week, all patients, including those who had suffered anaphylactic reactions and those who had only presented cutaneous manifestations, remained asymptomatic. CONCLUSIONS: Anisakis simplex-allergic patients tolerate ingestion of dead larvae. It is probable that these patients can eat frozen fish and that a seafood free diet is not necessary. PMID- 11436973 TI - Bezafibrate-induced anaphylactic shock: unusual clinical presentation. AB - We report a case of a patient who suffered generalized urticaria, chest tightness, wheezing, nausea, vomiting, hypotension, and loss of consciousness. Two hours earlier she had taken Eulitop Retard following lunch. She had tolerated all the implicated food after the reaction. Allergy evaluation revealed intense positive responses to intradermal tests with bezafibrate active component and Eulitop Retard (skin tests in control subjects were negative). Specific IgE tests (RAST) to Eulitop Retard were negative. An IgE mechanism is suggested to be responsible for this adverse reaction on the basis of the positive skin tets. The delayed onset (two hours) of this anaphylactic shock is unusual. Although infrequent, it may be caused by the specific pharmacokinetic characteristics of this drug, which is a slow releasing agent, mainly absorbed in the gut. The drug was taken just after lunch, and this concomitant food ingestion could also have produced a delay in gastric drainage and a retarded drug absorption. An IgE mediated accelerated type reaction could also explain this delay. Apparently the patient reacted after the first contact to the drug, and the absence of a sensitization period is not usual in this type of immune reponse. Finally, we recommend the performance of prick and intradermal skin tests prior to any systemic challenge when allergic reactions to fibric acid derivatives are suspected. PMID- 11436974 TI - Menthol-induced asthma: a case report. AB - A case of asthma due to menthol is reported in a 40-year-old woman with no history of asthma or any other allergy. During the last two years, the patient had presented dyspnea, wheezing and nasal symptoms when exposed to mentholated products such as toothpaste and candies. The etiology was suggested by the history of exposure and diagnosis was established by skin tests and bronchial challenge with menthol. The patient achieved control of symptoms by avoiding menthol and its derivatives. PMID- 11436975 TI - Gradient micropattern immobilization of heparin and its interaction with cells. AB - Heparin was immobilized on a polystyrene plate in a graded micropattern by photolithography. The micropattern was confirmed by staining with brilliant green. In the presence of basic fibroblast growth factor (bFGF), the growth of NIH3T3 cells was enhanced only in the heparin-immobilized regions. This result indicated that gradient-micropattern-immobilized heparin activated bFGF for cell growth. In addition, the growth of cells was found to be affected by the surface density of immobilized heparin. The surface density was regulated by a gap length of 2 microm-width stripes immobilized with heparin. Although a high density (a region having short gap length) of immobilized heparin suppressed the cell growth in the absence of bFGF, it enhanced cell growth in the presence of bFGF. The dependence of cell function on the density of immobilized heparin was visualized by gradient-micropattern-immobilization. PMID- 11436976 TI - A review of factors affecting the performances of latex agglutination tests. AB - The present review describes the different strategies followed to improve the performance of latex agglutination tests. The analysis is mainly focused on the diverse parameters that affect the final colloidal stability of the immunoprotein latex system. These parameters include: the surface properties of polymer carriers; the different kind of antibodies usually employed; the use of BSA as stabilizer; the co-adsorption of various macromolecules (BSA, surfactants and lipids) and antibodies; recent approaches to colloidal stability at high ionic strengths due to hydration forces; and the covalent coupling of antibodies on functionalized latex particles. Special emphasis is given to the relation between electrophoretic mobility and the colloidal stability of the sensitized particles and how this knowledge can be utilized for a better understanding of the immunoagglutination kinetic. PMID- 11436977 TI - Assembly properties and applications of a new exopolymeric compound excreted by Pseudoalteromonas antarctica NF3. AB - The self assembly properties and applications of an exopolymeric compound (EC) of a glycoprotein character excreted by a new gram-negative species, Pseudoalteromonas antarctica NF3, have been reviewed. This compound exhibited surface-active properties in water, with a concentration of 0.20 mg ml(-1) being the key value associated with its physicochemical properties. Unsonicated EC aqueous dispersions showed the coexistence of concentric multilamellar and small unilamellar aggregates by transmission electron microscopy (TEM). Sonication of these dispersions revealed that each lamellae of the initial multilamellar structures were made up of various subunits coiled coils. As for the ability of this exopolymeric biomaterial to coat phosphatidylcholine (PC) liposomes and to protect these vesicles against different surfactants, freeze-fracture TEM micrographs of liposome/EC aggregates revealed that the addition of the EC to liposomes led to the formation of a film (polymer adsorbed onto the bilayers) that coated very well the PC bilayers. The complete coating was already achieved at a PC:EC weight ratio of about 9:1. An increasing resistance of PC liposomes to surfactants (in particular sodium dodecyl sulfate) occurred as the proportion of EC in the system rose, although this effect was more effective at low EC proportions (PC:EC weight ratios from 9:1 to 8:2). Although a direct dependence was found between the growth of the enveloping structure and the resistance of the coated liposomes to be affected by the surfactants, the best protection occurred when this structure was a thin film of about 20-25 nm formed by nine to ten layers of about 2-3 nm. PMID- 11436978 TI - Preparation of poly(D,L-lactide) nanoparticles assisted by amphiphilic poly(methyl methacrylate-co-methacrylic acid) copolymers. AB - When co-precipitated with amphiphilic copolymers from DMSO, poly(D,L-lactide) (PLA) can be readily converted into stable sub-200 nm nanoparticles by addition of an aqueous phase, free of any polymeric stabilizers such as poly(vinyl alcohol) or Poloxamer. In this work, the ability of random poly(methyl methacrylate-co-methacrylic acid) copolymers (PMMA-co-MA) to stabilize PLA nanoparticles was demonstrated, and the properties of PLA/PMMA-co-MA nanoparticles were investigated. When co-precipitated with PMMA-co-MA, PLA was totally converted into nanoparticles using a polymer concentration in DMSO (Cp) below 17.6 mg ml(-1), and a PMMA-co-MA proportion above a critical value depending on the content of MA repeating units (X). For instance, the lowest PMMA co-MA proportion required was 0.9 mg mg(-1) PLA for X = 12%, and 0.5 mg mg(-1) PLA for X = 25% (for C(PLA) = 16 mg ml(-1) DMSO). The nanoparticle diameter was essentially independent of X, the proportion of PMMA-co-MA, and the PLA molecular weight, except for oligomers for which the nanoparticle diameter was smaller. It decreased when the organic phase was diluted (126 +/- 13 nm for Cp = 17.6 mg ml( 1), and 81 +/- 5 nm for C(P) = 5.6 mg ml(-1)). The time-dependence of the stability and the degradation of PLA/PMMA-co-MA nanoparticles was discussed. One of the main advantages of this technique is the ability to control surface properties and to bring functional groups to otherwise non-functionalized PLA nanoparticles. To illustrate this, a conjugate of PMMA-co-MA25 and biotin was synthesized, and used to prepare biotinylated nanoparticles that could be detected by fluorescence and transmission electron microscopy after infiltration into ligatured rat small intestine. PMID- 11436979 TI - Interaction of poly(2-acrylamido 2-methylpropane sulfonate)-grafted polystyrene beads with cationic complement proteins. AB - Influence of various biomaterials on the complement system in serum has been intensively studied by many research groups, since activation of the complement pathway in vivo has been known to give rise to some pathological conditions, such as inflammation and anaphylaxis. Much effort has been devoted to develop new materials that do not activate or deteriorate the complement system. The present work is aimed at revealing the mode of reactions of anionic poly(2-acrylamido 2 methylpropane sulfonate) grafted on polystyrene bead (PAMPS-g-bead) with serum complement. Complement activity assay, determination of complement proteins levels, and immunoblot analysis were carried out for sera pretreated with PAMPS-g beads. The results clearly showed that, when PAMPS-g-beads were incubated with serum, those beads adsorbed several complement proteins, i.e. C1q, factor D, factor P, C6, and C8, but the generation of activation fragments of complement components was not observed. Especially, factor D was most effectively removed from serum, resulting in potential inhibition of the alternative pathway. A larger amount of PAMPS-g-beads was needed to decrease the serum CH50 level. That may be caused by removal of C6. Although some polyanions, such as dextran sulfate, were reported to activate the complement system, the obtained results indicate that the PAMPS-g-bead is not an activator of the complement pathway, but acts as an adsorbent of complement components. One possible clinical application of the PAMPS-g-beads is adsorption of serum factor D by extracorporeal treatment of patients with renal failure with a high level of factor D, because the increased quantity of factor D in serum may cause consistent activation of the alternative pathway. PMID- 11436980 TI - Effects of calmodulin antagonist (W-7) on phorbol ester (PMA)-induced contractile response in isolated rat aorta. AB - The aim of this study was to investigate effects of calmodulin antagonist (W-7) on the contractile response of the rat aorta induced by activation of protein kinase C (PKC) by phorbol ester. Phorbol 12-myristate 13-acetate (PMA) produced biphasic contraction i.e., a sustained contraction (initial contraction) and 17.9 +/- 1.7 min later, this progressively developed contraction was changed to a delayed contraction superimposed on the initial contraction. The delayed contraction was completely inhibited by treatment with nicardipine. The onset of the delayed contraction was significantly delayed by treatment with W-7, whereas same concentration of W-7 showed a weak relaxant effect (10%) on the PMA-induced maximal contraction of aorta. Higher concentration of W-7 strongly inhibited PMA induced sustained contraction. These results suggest that PMA-induced biphasic contractile response may be regulated by calmodulin. PMID- 11436981 TI - Molecular cloning and sequencing of myosin light chains in human megakaryoblastic leukemia cells. AB - Myosin light chain genes of hematopoietic cells have yet to be characterized. We cloned the full-length cDNAs of 20 kDa regulatory myosin light chain (MLC-2) and 17 kDa essential myosin light chain (MLC-3) from Meg-01, a human megakaryoblastic leukemia cell line. Both MLC-2 and MLC-3 gene are transcribed ubiquitously in various hematopoietic cells. The MLC-2 open reading frame of 516 nucleotides encoding a protein of 172 residues was detected in cloned cDNA of 967 nucleotides. The Ca2+-binding domain and five phosphorylation sites were highly conserved. The deduced amino acid sequence has a 99.4% and 100% homology with that of human fetus brain and human lymphocyte, respectively. The MLC-3 open reading frame of 453 nucleotides encoding a protein of 151 residues was detected in cloned cDNA of 742 nucleotide. The MLC-3 protein is 99.3% identical to that of human fibroblasts. These results suggest that hematopoietic myosin light chain proteins are similar to those of other nonmuscle cells and smooth muscle, thus differing from skeletal and cardiac muscles. PMID- 11436982 TI - Difference in effects of stretch on depressive effect of endothelium-derived nitric oxide on noradrenaline- and high-K+-induced contractions between the aortae from normotensive and spontaneously hypertensive rats. AB - Difference in effects of stretch tension on endothelium-derived nitric oxide (EDNO)-dependent depression of noradrenaline (NA)- and high-K+-induced contraction between the aortae from normotensive Wistar Kyoto rats (WKY) a nd stroke-pronespontaneously hypertensive rats (SHRSP) was studied. NA-induced contraction in preparations both from WKY and SHRSP was augmented in the presence of N(omega)-nitro-L-arginine (L-NNA). This augmentation was minimized when the spontaneous tone, which was more prominent in preparations from SHRSP, was subtracted and the effects of L-NNA became less prominent in preparations from SHRSP. The effects of L-NNA were maximal at the stretch tension of 15 mN and, then, decreased as stretch tension increased in both preparations when the spontaneous tone was subtracted. The effects of L-NNA were less prominent when the contraction was initiated by high-K+, although the effects of stretch on high K+-induced contraction were similar to that of NA-induced contraction. These results suggested 1) that both NA- and high-K+-induced contractions are depressed by EDNO, 2) that the release of EDNO induced by high-K+ is less than that by NA, 3) that increase in stretch tension decreases the release of EDNO, and 4) that the depressive effect of EDNO on contraction is impaired in the aorta of SHRSP. PMID- 11436983 TI - Morphological changes and recovery process in the tenotomized soleus muscles of the rat. AB - Tenotomized soleus muscles of adult rats were analyzed morphologically and biochemically with special reference to the recovery process. Light microscopic observations of semi-thin sections showed that the characteristic central core lesion was most extensive at 1 week after tenotomy and began to diminish in extent at 2 weeks until no trace of lesion could be seen by 6th week, as confirmed by thin-section electron microscopy. Three phases of changes in the cross-sectional area of muscle fibers after tenotomy were demonstrated by morphometry: phase I designated as the initial increase up to the 3rd day, phase II as the progressive decrease until the 4th week, and phase III as the recovery to normal or even hypertrophy. In electron microscopy, the earliest alteration of myofibrils was recognized at 3 days after tenotomy. The Z discs showed a wavy or zigzag profile with frequent longitudinal splitting of myofibrils. From the 2nd week on, muscle fibers underwent a process of recovery, replacing the central core lesion with new myofibrils in which a reassembly of thick filaments into bundles of thin filaments took place, with Z discs being aligned adjacent to the peripheral complete myofibrils. In SDS-polyacrylamide gel electrophoresis, the molar ratio of myosin to actin diminished markedly as the central core lesion developed and gradually returned to normal with time, correlating well with the loss and subsequent reassembly of thick filaments. PMID- 11436984 TI - Effects of pinealectomy and sham-surgery on the area postrema in rats: a quantitative histological study with special reference to capillaries and neuronal cell nuclei. AB - This study aimed to clarify the effects of pinealectomy and sham-surgery on the area postrema (AP) by quantitative histological methods. Male, Wistar rats of normal (NO), sham-operated (SX), and pinealectomized (PX) groups were used in the late dark phase at 7 weeks of age. Consecutive frontal sections including the AP were stained with hematoxylin and eosin, and immunostained using PGP 9.5 for neurons, or GFAP or vimentin for glial cells. Consecutive sections of the AP were separated into five portions starting from the point of the central canal opening to the fourth ventricle in the caudal direction, and used for measurements. Mean cross-sectional areas of capillaries showed a lower value in the SX group than in the other two groups (vs NO, P<0.005; vs PX, P<0.03). In addition, the frequency distributions of the nuclear diameters of nerve cells showed different patterns among the three experimental groups (P<0.01), the frequency of large nuclei being higher in the SX group than in the other two groups. Possible mechanisms of the effects of sham-pinealectomy and pinealectomy and significance of the pineal-AP relation are discussed. The results of this study indicate that stuctural changes in the AP can be induced by intracranial surgery, suggesting certain pineal involvement in these changes. PMID- 11436985 TI - Development of astrocytes in the mouse hippocampus as tracked by tenascin-C gene expression. AB - Tenascin-C (TN-C) is an astroglia-derived extracellular matrix protein that has been shown to be an early marker for astroglial precursors in the embryonic mouse brain. This study examined astroglial generation, migration, and differentiation in the developing mouse hippocampus by in situ hybridization histochemistry for TN-C mRNA. Special reference was given to the difference in the mode of astroglial development between the two cortical structures of the hippocampus: the dentate gyrus and Ammon's horn. TN-C-positive cells were found in the ventricular germinative zone of the hippocampus as early as the 15th gestational day, and the labeled cells in the zone apposed to the fimbria migrated tangentially through the subpial area towards the forming dentate gyrus. The TN-C positive cells aligned in the dentate gyrus exhibited the characteristic morphology of unipolar astrocytes as revealed by double labeling with glial fibrillary acidic protein (GFAP)-immunohistochemistry. On the other hand, the TN C-positive cells ranging over a wide area of the ventricular germinative zone facing the forming Ammon's horn migrated radially towards the cortex, with most of them aligned in the Ammon's horn exhibiting a GFAP-positive stellate morphology. The onset of migration towards the dentate gyrus was two days earlier than that towards the Ammon's horn. TN-C-positive cells in both cortical structures exhibited a DNA-replicating activity after settlement in the early postnatal stage and were considered to further generate astrocytes. On the other hand, TN-C-positive cells with DNA-replicating activity were also found in the subpial migratory stream moving towards the dentate gyrus and were considered to form the subpial matrix for the generation of the dentate astrocytes. Migratory TN-C-positive cells directed towards both the dentate gyrus and Ammon's horn were apposed to radial glial processes and were believed to be guided by contact with these processes in a manner similar to migratory immature neurons. These findings indicate that TN-C-positive cells for the dentate gyrus and those for the Ammon's horn have different migratory patterns and undergo different morphological differentiations depending on their site of origin at the early stage of astrogliogenesis and corresponding to the different modes of neurogenesis in the two cortical structures. PMID- 11436986 TI - Immunolocalization of aquaporin-8 in rat digestive organs and testis. AB - Expression of aquaporin-8 mRNA has previously been shown in hepatocytes, pancreatic acinar cells, colon epithelial cells and seminiferous tubules of the testis in the rat by in situ hybridization technique. However, immunolocalization of this water channel has not yet been demonstrated. In the present study, the localization of immunoreactive aquaporin-8 and expression of the mRNA were examined in rat organs (cerebrum, cerebellum, eye, salivary gland, heart, lung, liver, pancreas, esophagus, stomach, jejunum, ileum, colon, testis, ovary, kidney, spleen and lymphnode) by immunohistochemistry using an antibody against aquaporin-8 and ribonuclease protection assay. Aquaporin-8 was distinctly immunolocalized on the apical membranes of pancreatic acinar cells and mucosal epithelium of the colon and jejunum. In the liver, the bile canalicular membrane of hepatocytes was immunostained. In the testis, immunoreactive aquaporin-8 was demonstrated on the luminal side of the seminiferous tubules. At high magnification, the peroxidase reaction products appeared on the ramified cytoplasmic membrane of Sertoli cells surrounding the residual bodies or spermatogenic cells. Specificity of the antibody was verified by Western blot analysis showing a minor approximately 28 kDa band (deduced deglycosylated form of aquaporin-8) and a major approximately 30 kDa band (glycosylated form) in these organs. The intensity of aquaporin-8 immunoreactivity was approximately comparable to that of aquaporin-8 mRNA expression in the liver, pancreas, colon, jejunum and testis. The aquaporin-8 mRNA expression in the hepatocytes was presumed to be closely associated with the structure of bile canaliculi since the message was detected in hepatocytes immediately after isolation from the liver but not in cells following cultivation for three days. The localization of immunoreactive aquaporin-8 indicated functions for this water channel in the secretion of bile and pancreatic juice, and the secretion or absorption of water in the colon and jejunum, and the maturation or liberation of spermatogenic cells in the testis. PMID- 11436987 TI - Role of heme oxygenase-1 and Kupffer cells in the production of bilirubin in the rat liver. AB - Heme oxygenase (HO)-1, the heme-degrading enzyme in macrophages, plays a key role in bilirubin metabolism. HO-1 is expressed in various tissue macrophages, especially Kupffer cells. This study aimed to examine the roles of macrophages and HO-1 in the modulation of heme catabolism in rat livers. Rats treated with or without liposome-encapsulated dichloromethylene diphosphonate, a macrophage depleting reagent, were administered with heat-denatured red blood cells (h-RBC), and the time course of the biliary output of bilirubin and the expressions of HO 1 mRNA and protein were monitored. Immunohistochemistry in the control rat liver revealed that Kupffer cells constitute a major cellular component expressing HO 1, while hepatocytes exhibited little expression. The levels of HO-1 expression in Kupffer cells were elevated immediately after injection of h-RBC. In Kupffer cell-depleted livers, however, HO-1-expressing cells were not detected even after h-RBC administration. HO-1 mRNA levels were elevated at 2 h after administration of h-RBC in control rat livers, while they were very low in Kupffer cell-depleted rat livers. The control and Kupffer cell-depleted groups exhibited distinct time courses of biliary bilirubin excretion. In the untreated control rats, total bilirubin excretion increased about two-fold at 5 h after h-RBC administration. In contrast, the Kupffer cell-depleting treatment decreased the level of bilirubin production; administration of h-RBC to Kupffer cell-depleted rats did not accelerate the generation of bilirubin. These results suggest that Kupffer cells serve both as a sensor for scenesent RBC clearance and an effector that upregulates heme-degrading capacity and bilirubin production. PMID- 11436988 TI - Three-dimensional analysis of nephrogenesis in the neonatal rat kidney: light and scanning electron microscopic studies. AB - In order to clarify the process of renal development more precisely than previously, the present study observed the rat neonatal kidney by scanning electron microscopy (SEM) of KOH digested tissue as well as by light microscopy of plastic sections. In the subcapsular region, aggregation of the mesenchymal cells was closely associated with the upper side of the ureteric duct ampulla. These mesenchymal cells projected a number of fine irregular processes at the basal portion facing the ureteric duct. A spherical cluster transformed from the mesenchymal cell aggregation was found on the lower side of the terminal ampulla, and was differentiated into the renal vesicle. Some cells at the top of the renal vesicle formed a cone-shaped projection and invaded the ureteric duct ampulla, forming a connection with it. In the advanced stage, a shallow transverse cleft appeared on the outer lateral side of the renal vesicle, and a second cleft was formed on the opposite side close to the junction between the renal vesicle and the ampulla. As the two clefts deepened, the vesicle assumed the well-known S shaped body. In the advanced S-shaped body, the lower limb became cup-shaped, while the segment between the middle and lower limbs of the "S" elongated to form a tubular structure (i.e., the prospective proximal tubule and Henle's loop). The upper limb of the "S" also increased its length to form a distal tubule. The middle limb of the "S", however, was attached firmly to the cup-shaped lower limb (i.e., the prospective renal corpuscle) and was considered to become the macula densa of the mature nephron. In the maturing renal corpuscle, irregularly shaped cells were observed as a sheet-like aggregation at its vascular pole and were continuous with the vascular smooth muscle cells. These findings will help toward a better understanding of the morphological complexities of nephrogenesis. PMID- 11436989 TI - Morphological characteristics of Schwann cells in the islets of Langerhans of the murine pancreas. AB - The present study demonstrated the three-dimensional architecture of peri-insular nerve plexuses in the murine pancreas by the combined use of light microscopy of S-100 immunostained sections, transmission electron microscopy (TEM) of thin sections, and scanning electron microscopy (SEM) of KOH digested tissues. By light microscopy of thin sections immunostained with anti-S-100 antibody, Schwann cells were often found on the margin of the islets as if delimiting the islet and exocrine parenchyma. In thick sections, Schwann cells of the islet connected their thin and slender processes with each other to form a delicate network on the surface of the islet. By TEM, Schwann cells were observed as an attenuated sheet that invested the surface of the islet. Axon terminals were usually found on the outer surface of these membranous Schwann cells. SEM of KOH digested tissues revealed that nerves reaching the islet spread on the insular surface. Schwann cells in this portion extended their thin membranous processes, which directly covered the basal part of several endocrine cells as a whole. Numerous axons with varicosities were usually found on the surface of these membranous Schwann cells, but sometimes crept beneath them. These findings indicate that "the interstitial cells" described by light microscopists are peculiar-shaped Schwann cells present in the islets. The functional significance of the rich innervation of the islets is also briefly discussed in the present study. PMID- 11436990 TI - Purkinje cells in the adult cat cerebellar cortex possess a perineuronal net of proteoglycans. AB - The Purkinje cells in the adult cat cerebellar cortex were found to possess perineuronal proteoglycans which could be stained with our fine cationic iron colloid and Fujita's highly concentrated aldehyde fuchsin, and digested by chondroitinase ABC/keratanase/ heparitinase and hyaluronidase. The Purkinje cells are surrounded by some collagenous elements which are stained with Gomori's ammoniacal silver and digested by collagenase. The Purkinje cells also express nerve cell surface glycoproteins which are labeled with lectin Vicia villosa agglutinin and digested by a double treatment with collagenase and endo-alpha-N acetylgalactosaminidase. Sole digestion by endo-alpha-N-acetylgalactosaminidase never erased the lectin labeling of the nerve cell surface glycoproteins. These findings suggest that the collagenous elements mediate the linkage of the perineuronal proteoglycans to the nerve cell surface glycoproteins. It is presumed that in mice and rats, the perineuronal nets of proteoglycans and nerve cell surface glycoproteins of the Purkinje cells are so thin or coarse that they can not be sufficiently visualized under the light microscope. PMID- 11436991 TI - Effects of photoperiod and melatonin on the development of growth hormone cells and the pituitary-adrenal axis in the Djungarian hamster, Phodopus sungorus. AB - The development of GH cells and the pituitary-adrenal axis was morphologically examined in male Djungarian hamsters (Phodopus sungorus) exposed to short days and those kept under long days and receiving daily afternoon injections of melatonin, from the time of weaning (20 days) until 100 days of age. The postnatal increase in area of ACTH cells under long days was inhibited in short day-exposed or melatonin-treated animals. It was suggested that a short photoperiod may suppress, via melatonin, the development of ACTH cells. GH cells were not affected by age, photoperiod or exogenous melatonin. Under long days, the zona fasciculata decreased in volume with age, while the zona reticularis increased. Such changes in the volumes of these adrenocortical zones were depressed under short days. In addition, the volumes of the zona fasciculata and zona reticularis in long-day-housed animals became respectively larger and smaller subsequent to orchidectomy and melatonin administration. These results suggest that fasciculata cells in deeper levels become progressively differentiated into reticularis cells, that short photoperiod inhibits development of both zonae, and that such an inhibition is caused mainly by the decreased secretion of androgens. PMID- 11436992 TI - Asialoglycoprotein receptors on rat dendritic cells: possible roles for binding with Kupffer cells and ingesting virus particles. AB - Rat dendritic cells selectively bind to Kupffer cells in vitro. The present study aimed to reveal adhesion molecules on dendritic cells and their roles in the host defense system. The in situ binding assay to examine the effects of pretreatment of dendritic cells with various kinds of monosaccharides suggested that N acetylgalactosamine was necessary for the binding of dendritic cells to Kupffer cells. This binding was also attenuated when dendritic cells were injected into an ex vivo liver perfusion circuit together with N-acetyl-galactosamine. It was further shown that the majority of rat lymph dendritic cells and some interdigitating dendritic cells in the lymph nodes possessed asialoglycoprotein receptors specific for N-acetylgalactosamine/galactose as detected by immunostaining. Lymph dendritic cells could ingest virus particles in vitro, even though these cells showed no phagocytic activity for latex particles. The results indicate that rat dendritic cells possess asialoglycoprotein receptors which are probably utilized to recognize Kupffer cells for their recruitment to the liver and possibly to recognize virus particles prior to phagocytosis. PMID- 11436993 TI - A conceptual model for describing decision-making situations in integrated natural resource planning and modeling projects. AB - A conceptual model is developed herein for the purpose of stimulating discussions within groups planning and carrying out integrated natural resource projects. We first describe four basic components of integrated planning and modeling efforts: people, databases, technology, and organizational commitment. Second, we provide one view of the relationship between the size of the project's decision-making body and the timing of decisions during a project's life cycle. Finally, these two discussions are combined into a conceptual model describing the dynamic nature of decision-making within integrated projects. The abstractions and generalizations described here are not unique to private industry or governmental organizations and should provide the basis for a discussion of decision-making issues among interdisciplinary professionals embarking on large-scale or complex modeling efforts. PMID- 11436994 TI - High-frequency zone of river desiccation disasters in China and influencing factors. AB - In recent years, the desiccation of the lower Yellow River has raised concerns in the government, public, and scientific community in China. Long-term and widespread desiccation of rivers is a disaster with many adverse environmental implications. It has been found in this study that there exists a high-frequency zone of river desiccation disasters at 34 degrees -42 degrees N in the North China Plain. The hazardous environment is characterized by semiarid climate, widely distributed thick loess in the basin and a "hanging river bed" in the plain as well as unfavorable man-water-land coupling relationships. In this setting, the sharply increased water diversion by man since the late 1950s led to the occurrence of river desiccation disasters in the lower reaches of the river in this area. PMID- 11436995 TI - Indicators for transboundary river management. AB - The aim of this paper is to analyze the potential of indicators for integrated river basin management and to develop a set of indicators for the management of transboundary river basins. An indicator, comprising a variable or some aggregation of variables, describes a system or process such that it has significance beyond the face value of its components. Integrated river basin management takes into account policies and measures for the multifunctional use of rivers on a catchment scale and associated institutional changes. Indicators are useful instruments for this process for two reasons. Firstly, they meet the information need of policy- and decision-makers. Secondly, indicators can be used to structure the definition and description of information needs and collection of information between the different international, institutional, and sectoral management levels. The development of indicators involves a number of steps: definition of aim, construction of conceptual model, selection of variables, comparison with selection criteria, database assessment, and indicator selection. In this paper these steps are discussed and specified for integrated river basin management. This results in a set of indicators describing the pressure to the river, the state of the river ecosystem, the impact to goods and services provided by the river, and the societal response. The proposed set of indicators measured at a river basin scale provides integrated information on the use and supply of goods and services, underlying cause-effect relationships and possible trade-offs and their spatial distribution (e.g., upstream versus downstream). Furthermore, we propose a division of tasks and responsibilities for river basin management with regard to the development of indicators, data collection, and their application in decision-making. PMID- 11436996 TI - The environmental impact of shrimp aquaculture: causes, effects, and mitigating alternatives. AB - Attracted by the demand for shrimp in the developed countries, shrimp aquaculture has expanded rapidly, mainly in the subtropical and tropical lowlands of America and Asia. This work provides a global review and viewpoint on the environmental impacts of shrimp aquaculture, considering the causes and effects of the siting and operation of shrimp ponds and abandonment of farm facilities. Additionally, mitigating alternatives are discussed. To date, approximately 1-1.5 million ha of coastal lowlands have been converted into shrimp ponds, comprising mainly salt flats, mangrove areas, marshes, and agricultural lands. The impact of shrimp farming of most concern is the destruction of mangroves and salt marshes for pond construction. Compatibility with other users, the presence of buffer zones, maintaining an acceptable balance between mangroves and shrimp pond area, improved pond design, reduction of water exchange, and an improved residence time of water, size and capacity to assimilate effluents of the water body, are examples of ways to mitigate the adverse effects. The use of mangroves and halophytes as biofilters of shrimp pond effluents offers an attractive tool for reducing the impact in those regions where mangrove wetlands and appropriate conditions for halophyte plantations exist. Healthy seed supply, good feed with the use of prophylactic agents (including probiotics), good water quality, and lower stocking densities are examples of actions suggested to control disease in shrimp farming. Finally, in the context of integrated management, research priorities are suggested. PMID- 11436997 TI - Urban environmental management in Botswana: toward a theoretical explanation of public policy failure. AB - For the last three and half decades, Botswana has been widely acclaimed to be one of sub-Saharan Africa's longest and most stable liberal democracies, coupled with and sustained by a growing economy. One of the major contradictions, however, within this development scenario, has been the neglect of environmental problems in the country in general, and urban environmental issues in particular. Part of the problem fueling the misconception of environmental issues in Botswana is the state's domination of the country's environmental agenda. This is linked to the power disjunction in decision-making and policy processes between state and nonstate actors over the most appropriate course of action to tackle the problems. Without adopting appropriate analytical frameworks, it is possible that the problem of urban environmental mismanagement in Botswana will persist. This article examines some of the major urban environmental issues in Botswana from neighborhood, citywide, and urban-rural interface perspectives. Further, the elite theory of public policy is used to explain constraints on policy change in the urban environmental management arena in Botswana. PMID- 11436998 TI - Integrating nonindigenous aquatic plant control with protection of snail kite nests in Florida. AB - The endangered snail kite (Rostrhamus sociabilis) feeds primarily on the freshwater apple snail (Pomacea paludosa) in Florida. The nonindigenous, floating water hyacinth (Eichhornia crassipes) and water lettuce (Pistia stratiotes) impede kites from finding snails. Effective control of these aquatic plants in the littoral zone of central and south Florida lakes benefits kites by maintaining open foraging habitat. However, incidental herbicide spraying of nesting substrates result in nest collapse when kites breed in nonwoody, emergent plants [cattail (Typha spp.) and giant bulrush (Scirpus validus)] in the outer littoral zone during lower lake levels. Many endangered species recovery plans and their implementation have experienced problems due to inaction and/or noncooperation by various governmental agencies and their personnel. Herein, we describe the development and implementation of a buffer zone strategy to prevent secondary impacts from an aquatic plant control program to snail kites nesting on lakes in central and south Florida. A strategy was jointly developed by personnel of five state and federal agencies to control herbicide application near kite nesting areas during the normal breeding season. Although requiring various modifications during its implementation, this cooperative effort successfully integrated aquatic plant control objectives with snail kite conservation on Lake Okeechobee during 1988. The program was expanded the following year to lakes Kissimmee and Tohopekaliga. Since the implementation of the snail kite impact preclusion program, no nest loss was attributed to incidental herbicide applications on lakes Okeechobee, Kissimmee, and Tohopekaliga. PMID- 11436999 TI - Trends in vegetation degradation in relation to land tenure, rainfall, and population changes in Peddie district, Eastern Cape, South Africa. AB - Spatial and temporal variations in vegetation are examined in relation to land tenure, population increase, and rainfall variation in a part of Peddie district, Eastern Cape. Sequential aerial photographs between 1938 and 1988 are analyzed to determine trends in vegetation and population change in three different land tenure units. The areal extent at each date of four distinct vegetation categories is determined using PC ARC/INFO GIS. Long-term annual rainfall trends for the area are analyzed and juxtaposed with vegetation changes. Extensive ground-truthing exercises are carried out to verify the present condition of vegetation condition in terms of cover and species composition. Differences in land-tenure systems are discerned as the dominant factor controlling variations in vegetation degradation. The study also reveals that neither population changes nor rainfall variations can explain the observed trends in vegetation degradation. Earlier injudicious land-use practices, sustained since the turn of the last century, may provide plausible explanations for the trends and present status of vegetation degradation in the area. PMID- 11437000 TI - Stakeholder management for conservation projects: a case study of Ream National Park, Cambodia. AB - The paper gives an account of the development and implementation of a stakeholder management framework at Ream National Park, Cambodia. Firstly, the concept of stakeholder is reviewed in management and in conservation literatures. Secondly, the context in which the stakeholder framework was implemented is described. Thirdly, a five-step methodological framework is suggested: (1) stakeholder analysis, (2) stakeholder mapping, (3) development of generic strategies and workplan, (4) presentation of the workplan to stakeholders, and (5) implementation of the workplan. This framework classifies stakeholders according to their level of influence on the project and their potential for the conservation of natural resources. In a situation characterized by conflicting claims on natural resources, park authorities were able to successfully develop specific strategies for the management of stakeholders. The conclusion discusses the implications of the Ream experience and the generalization of the framework to other protected areas. PMID- 11437001 TI - A combined park management framework based on regulatory and behavioral strategies: use of visitors' knowledge to assess effectiveness. AB - In light of the increasing mandate for greater efficiency in conservation of natural reserves such as national parks, the present study suggests educational approaches as a tool to achieve conservation purposes. Currently, the management of human-wildlife interactions is dominated by regulatory strategies, but considerable potential exists for environmental education to enhance knowledge in the short run and to prompt attitude change in the long run. A framework for conservation based on both traditional regulatory- and behavior-oriented strategies was proposed whereby the level of knowledge that park visitors have acquired comprises an obvious outcome and establishes a basis upon which the effectiveness of regulatory- and behavior-based regimes could be assessed. The perceptions regarding park-related issues of two distinct visitor groups (locals and nonlocals) are summarized from a survey undertaken in Vikos-Aoos national park. The findings suggest a superficial knowledge for certain concepts but little profound understanding of the content of such concepts, indicating that knowledge-raising efforts should go a long way towards establishing a positive attitude for the resource. Visitors' poor knowledge of the park's operation regulation contest the efficiency of the presently dominant regulatory management regime. While geographical distances did not appear to significantly differentiate knowledge between the two groups, wilderness experience (as certified by visits to other parks) was proved to be an impetus for generating substantial learner interest in critical park issues among nonlocal visitors. School education and media were found to be significant knowledge providers. PMID- 11437002 TI - Using force analysis to target collection and analysis of environmental information. AB - Knowledge of the forces driving and modifying ecosystems can be employed in concert with signal analysis to target the data most likely to yield sensitivity and resilience information. One can optimize return of information per investment of resources by targeting segments of signals that are dominated by the force of interest, coupled with scientific understanding of the system of interest. This force analysis approach is an effective means both to design efficient new monitoring programs and to target relevant information in large data files. We present five example applications of force analysis. Three examples illustrate this approach for an evaluation of whether Canadian rivers might be sensitive to changes in climate. It was concluded that Canadian rivers appear to be sensitive to changing climate. A fourth example illustrates how automated snow pillow data may be evaluated to ascertain the sensitivity of snow accumulation to change in climate. It was concluded that snow accumulation at the site evaluated did appear to be sensitive to changing climate. The fifth example illustrates the assessment of whether a river recovers with the elimination of inputs of iron from an abandoned mine. It was concluded that resilience remained unproven since the river had not as yet restabilized. The force analysis approach focuses data collection or data evaluation on those data required to answer specific resource management questions, greatly reducing collection or consideration of data that are not relevant to that question. This approach is potentially very cost efficient and therefore is likely to be of interest to hydrologists, climatologists, and environmental data managers. PMID- 11437003 TI - Effects of local land use on physical habitat, benthic macroinvertebrates, and fish in the Whitewater River, Minnesota, USA. AB - Best management practices (BMPs) have been developed to address soil loss and the resulting sedimentation of streams, but information is lacking regarding their benefits to stream biota. We compared instream physical habitat and invertebrate and fish assemblages from farms with BMP to those from farms with conventional agricultural practices within the Whitewater River watershed of southeastern Minnesota, USA, in 1996 and 1997. Invertebrate assemblages were assessed using the US EPA's rapid bioassessment protocol (RBP), and fish assemblages were assessed with two indices of biotic integrity (IBIs). Sites were classified by upland land use (BMP or conventional practices) and riparian management (grass, grazed, or wooded buffer). Physical habitat characteristics differed across buffer types, but not upland land use, using an analysis of covariance, with buffer width and stream as covariates. Percent fines and embeddedness were negatively correlated with buffer width. Stream sites along grass buffers generally had significantly lower percent fines, embeddedness, and exposed streambank soil, but higher percent cover and overhanging vegetation when compared with sites that had grazed or wooded buffers. RBP and IBI scores were not significantly different across upland land use or riparian buffer type but did show several correlations with instream physical habitat variables. RBP and IBI scores were both negatively correlated with percent fines and embeddedness and positively correlated with width-to-depth ratio. The lack of difference in RBP or IBI scores across buffer types suggests that biotic indicators may not respond to local changes, that other factors not measured may be important, or that greater improvements in watershed condition are necessary for changes in biota to be apparent. Grass buffers may be a viable alternative for riparian management, especially if sedimentation and streambank stability are primary concerns. PMID- 11437004 TI - Criteria for oil spill recovery: a case study of the intertidal community of Prince William Sound, Alaska, following the Exxon Valdez oil spill. AB - Marine intertidal organisms in Prince William Sound were exposed to crude oil following the TN Exxon Valdez oil spill in 1989. The intertidal communities were also subjected to mechanical disturbance during invasive oil spill remediation and cleanup efforts. Using monitoring data collected from 1989 to 1997, impacts and eventual recovery were assessed at oiled but uncleaned sites and oiled and cleaned study areas. A statistical model where recovery was defined as parallelism between the time profiles at control and oiled sites was evaluated. Statistical analysis and graphical presentations of the data suggest intertidal epibiota communities recovered from the oil spill by 1992 at the oiled sites and by 1994 at the oiled and remediated sites. Empirical data from the intertidal monitoring program supports the use of tests of parallelism in evaluating recovery and the need to avoid simply the comparison of sample means from control and oiled sites. PMID- 11437005 TI - Adderall and the FDA. PMID- 11437006 TI - Is 18 months too early for the CHAT? PMID- 11437007 TI - Neuroimaging and anorexia nervosa. PMID- 11437008 TI - The meaning of medication. PMID- 11437009 TI - Hallucinations and psychosis. PMID- 11437010 TI - Citalopram for OCD and Tourette's syndrome. PMID- 11437011 TI - Periodic catatonia in an adolescent. PMID- 11437012 TI - The past 50 years of child and adolescent psychiatry: a personal memoir. PMID- 11437013 TI - Behavioral phenotypes of genetic syndromes: a reference guide for psychiatrists. AB - OBJECTIVE: To review the literature on behavioral phenotypes of genetic syndromes, displaying the data as a reference guide for everyday practice. METHOD: A computerized search was performed for articles published in the past 10 years, and selected papers were surveyed. RESULTS: The behavioral phenotypes of 11 major genetic syndromes were reviewed including the following topics: genetic etiology, genetic counseling, physical features, medical problems, cognitive and behavioral profile, and psychopathology. The speculated correlation between the identified gene and the pathophysiology of the cognitive and behavioral features is discussed. CONCLUSIONS: Updated knowledge of behavioral phenotypes will help psychiatrists identify these conditions, refer the patient and his/her family for genetic diagnosis and counseling, make specific treatment recommendations, and contribute to research and syndrome delineation. PMID- 11437014 TI - Efficacy of paroxetine in the treatment of adolescent major depression: a randomized, controlled trial. AB - OBJECTIVE: To compare paroxetine with placebo and imipramine with placebo for the treatment of adolescent depression. METHOD: After a 7- to 14-day screening period, 275 adolescents with major depression began 8 weeks of double-blind paroxetine (20-40 mg), imipramine (gradual upward titration to 200-300 mg), or placebo. The two primary outcome measures were endpoint response (Hamilton Rating Scale for Depression [HAM-D] score < or = 8 or > or = 50% reduction in baseline HAM-D) and change from baseline HAM-D score. Other depression-related variables were (1) HAM-D depressed mood item; (2) depression item of the Schedule for Affective Disorders and Schizophrenia for Adolescents-Lifetime version (K-SADS L); (3) Clinical Global Impression (CGI) improvement scores of 1 or 2; (4) nine item depression subscale of K-SADS-L; and (5) mean CGI improvement scores. RESULTS: Paroxetine demonstrated significantly greater improvement compared with placebo in HAM-D total score < or = 8, HAM-D depressed mood item, K-SADS-L depressed mood item, and CGI score of 1 or 2. The response to imipramine was not significantly different from placebo for any measure. Neither paroxetine nor imipramine differed significantly from placebo on parent- or self-rating measures. Withdrawal rates for adverse effects were 9.7% and 6.9% for paroxetine and placebo, respectively. Of 31.5% of subjects stopping imipramine therapy because of adverse effects, nearly one third did so because of adverse cardiovascular effects. CONCLUSIONS: Paroxetine is generally well tolerated and effective for major depression in adolescents. PMID- 11437015 TI - Fluoxetine treatment for obsessive-compulsive disorder in children and adolescents: a placebo-controlled clinical trial. AB - OBJECTIVE: This study assesses the efficacy and tolerability of fluoxetine in the acute treatment of child and adolescent obsessive-compulsive disorder (OCD) during a 13-week, double-blind, placebo-controlled study. METHOD: Eligible patients aged 7 to 17 (N = 103) were randomized at a ratio of 2:1 to receive either fluoxetine or placebo. Dosing was initiated at 10 mg daily for 2 weeks, then increased to 20 mg daily. After 4 weeks of treatment, and again after 7 weeks of treatment, non-responders could have their dosage increased by 20 mg daily, for a maximum possible dosage of 60 mg daily. Primary measure of efficacy was improvement in OCD symptoms as measured by the Children's Yale-Brown Obsessive Compulsive Scale (CY-BOCS). All analyses were intent-to-treat. RESULTS: Fluoxetine was associated with significantly greater improvement in OCD as assessed by the CY-BOCS (p = .026) and other measures than was placebo. Fluoxetine was well tolerated and had a rate of discontinuation for adverse events similar to that of placebo (p = 1.00). CONCLUSIONS: Fluoxetine 20 to 60 mg daily was effective and well tolerated for treatment of OCD in this pediatric population. PMID- 11437016 TI - School-based behavioral treatment for social anxiety disorder in adolescents: results of a pilot study. AB - OBJECTIVE: To provide preliminary estimates of feasibility and effectiveness for school-based behavioral treatment in adolescents with social anxiety disorder. METHOD: Six adolescents with social anxiety disorder were treated in a 14-session group treatment program conducted at their school. Assessments were conducted at baseline and after treatment. RESULTS: All participants were classified as treatment responders (markedly or moderately improved). Half of the participants did not meet diagnostic criteria for social phobia after treatment. Clinician severity ratings, as measured by the Anxiety Disorders Interview Schedule for Children for DSM-IV: Child Version and the Liebowitz Social Anxiety Scale for Children and Adolescents (LSAS-CA), decreased significantly after intervention, with effect sizes of 2.5 and 1.8, respectively. All LSAS-CA scale scores decreased significantly after treatment. Self-reported social phobia symptoms on the Social Phobia and Anxiety Inventory for Children were not significantly reduced. Fear and avoidance ratings of the 10 most feared situations significantly decreased after treatment, with effect sizes of 1.5 for anxiety and 2.1 for avoidance. CONCLUSIONS: This study provides preliminary support for the promise of school-based behavioral intervention for treating social phobia in adolescents. The school environment may be a rich and innovative setting for implementation of behavioral treatment because this is the setting where adolescents with social phobia endure the most distress. PMID- 11437017 TI - Comorbidity in childhood anxiety disorders and treatment outcome. AB - OBJECTIVE: Psychiatric comorbidity is common in anxious children. The purpose of this study was to investigate the impact of comorbidity on treatment outcome in anxious children. METHOD: Participants were 173 children between the ages of 8 and 13 years who met primary DSM-III-R/DSM-IV diagnoses of separation anxiety disorder, overanxious disorder/generalized anxiety disorder, or avoidant disorder/social phobia assessed by the Anxiety Disorders Interview Schedule for Children (ADIS-C). The majority (79%) had at least one comorbid diagnosis. Participants were randomly assigned to cognitive-behavioral therapy or waitlist. Group differences in ADIS-C diagnoses were compared after treatment. Multiple parent and child self-report measures were used to measure symptoms as well. RESULTS: Pretreatment comorbidity was not associated with differences in treatment outcome: 68.4% of noncomorbid participants and 70.6% of comorbid participants were free of their primary diagnosis after treatment. Regarding parent and child self-report symptoms, multivariate analyses of variance revealed significant time (treatment) main effects, but no significant main effect for group (comorbid status) or time/group interaction. CONCLUSIONS: The cognitive behavioral treatment program was similarly effective in anxious children with and without comorbid disorders; both groups showed clinically significant reductions in pretreatment diagnoses and symptoms. PMID- 11437018 TI - Impact of comorbidity on a cognitive-behavioral group treatment for adolescent depression. AB - OBJECTIVE: Examine hypotheses concerning the negative impact of lifetime psychiatric comorbidity on participation in, and benefit from, a cognitive behavioral group treatment for depression in adolescents (e.g., greater severity at intake, less recovery and more recurrence, less participation in treatment). METHOD: Across two previous studies conducted between 1986 and 1993, 151 depressed adolescents (aged 14-18) were randomly assigned to one of three treatment conditions (two active treatments and a waitlist control) and followed for 24 months posttreatment. Forty percent of participants had one or more lifetime comorbid diagnoses at intake. RESULTS: Comorbid anxiety disorders were associated with higher depression measure scores at intake and greater decrease in depression scores by posttreatment. Overall lifetime comorbidity was unrelated to diagnostic recovery, but lifetime substance abuse/dependence was associated with slower time to recovery. Participants with attention-deficit and disruptive behavior disorders were more likely to experience depression recurrence posttreatment. Associations between comorbidity and participation or therapy process measures were nonsignificant. CONCLUSIONS: Although some outcomes were worse for some comorbid diagnoses, the reassuring overall conclusion is that the presence of psychiatric comorbidity is generally not a contraindication for the use of structured group cognitive-behavioral interventions for depressed adolescents. PMID- 11437019 TI - Nurses' and physicians' assessment of mother-infant mental health at the first postnatal visits. AB - OBJECTIVE: To examine the degree to which physicians and nurses use their first postnatal contact with mothers and babies to learn about their psychosocial strengths and problems. METHOD: Forty-two consecutively born infants and their mothers were observed during their initial postnatal visit with a public health nurse and their physician in Montreal. Both visits were audiotaped. Tapes were analyzed for the number of physical and psychosocial topics discussed during the visits. Observers also rated the professionals' communication skills. During a later home visit, mothers were given the Edinburgh Postnatal Depression Scale, the Symptom Checklist-90-Revised, and the Working Model of the Child Interview (WMCI). RESULTS: Nurses spent more time with families, discussed more psychosocial issues, and were rated to be more sensitive interviewers than physicians. They were also concerned about the psychological well-being and psychological difficulties of more mothers than were the physicians. In contrast, physicians were more sensitive to pregnancy complications in younger mothers. Mothers' satisfaction correlated with the number of psychosocial issues discussed by both professional groups. WMCI data suggest that recent non-Western immigrants are overrepresented among insecurely attached mothers. CONCLUSIONS: Nurses and physicians obtain different data from the same patients and should increase their collaboration. PMID- 11437020 TI - Prevalence of social-emotional and behavioral problems in a community sample of 1 and 2-year-old children. AB - OBJECTIVE: To examine the prevalence of infant-toddler social-emotional and behavioral problems and associations with social-emotional competence, interference in family life, and parental worry. METHOD: The sample consisted of 1- and 2-year-old children (mean [MN] age = 24.8 months) from the baseline survey of a representative sample of healthy births (N= 1,280). Parent questionnaires included the Child Behavior Checklist (CBCL/2-3), Parenting Stress Index Short Form (PSI/SF), and Infant-Toddler Social and Emotional Assessment social emotional competence scales, as well as questions about parental worry and family activities. RESULTS: Approximately 80% of eligible subjects participated. The weighted prevalence of parent-reported subclinical/clinical CBCL/2-3 scores was 11.8% for 2-year-olds. Approximately 6% of parents of 1- and 2-year-olds reported clinical-level scores on the PSI Difficult Child (PSI/DC) scale, which was included as a proxy for behavior problems among 1-year-olds, for whom measures were limited. Sex differences were not observed. CBCL/2-3 and PSI/DC scores were uniquely associated with economic disadvantage (relative risk = 1.89 and 2.24, respectively). Approximately 32% of 2-year-olds with subclinical/clinical CBCL2-3 scores had delayed social-emotional competence. Problems were also associated with parental worry about child behavior and interference in family activities. CONCLUSIONS: A significant need for early identification of emotional/behavioral problems in very young children is highlighted by associations with delayed competence and disruptions in family life that may further contribute to risk for persistent problems. PMID- 11437021 TI - Prevalence of pervasive developmental disorders in the British nationwide survey of child mental health. AB - OBJECTIVE: The prevalence of pervasive developmental disorders (PDD) is not well established and needs monitoring. The prevalence of PDD in the 1999 nationwide British survey of child and adolescent mental health was investigated. METHOD: A randomized, stratified sample of children (N= 12,529) aged 5 to 15 years was generated from the Child Benefit Register. Trained interviewers interviewed parents and youths aged 11 or older with a standardized diagnostic interview (Development and Well-Being Assessment), and questionnaire data (Strengths and Difficulties Questionnaire) were obtained from teachers and parents, who also completed self-report measures of psychological distress. Final diagnostic determination was achieved by a team of experienced clinicians using all data sources. RESULTS: A total of 10,438 (83%) interviews were conducted. There were 2 girls with Rett syndrome (weighted prevalence: 3.8/10,000 girls) and 27 children with other PDD (weighted prevalence: 26.1/10,000). Compared with children with a psychiatric disorder other than PDD, social but not behavioral problems were more frequent in the PDD group. Parents of children with PDD had higher rates of psychological distress than those from the two comparison groups. CONCLUSIONS: Consistent with other recent surveys, PDD rates are higher than those reported 30 years ago. The burden associated with PDD is very high. PMID- 11437022 TI - Behavioral and emotional problems in Chinese adolescents: parent and teacher reports. AB - OBJECTIVE: This study examined behavioral and emotional problems in Chinese adolescents. METHOD: A sample of 1,694 adolescents aged 12 to 16 years participated in this study in 1997. Parents completed the Child Behavior Checklist (CBCL), and teachers completed the Teacher's Report Form (TRF). RESULTS: For both parent and teacher reports, internalizing syndromes were scored higher in girls and externalizing syndromes were scored higher in boys. Scores on most of the CBCL and TRF subscales were higher for the older adolescents. The overall prevalence rates of parent- and teacher-reported behavioral problems were 23.1% and 19.2%, respectively. The eight cross-informant syndromes were highly comorbid, with a significant association across syndromes (mean odds ratio = 16.1 for CBCL and 22.5 for TRF). Correlations between parent and teacher reports were 0.51 for eight subscales and 0.68 for Total Problems. CONCLUSIONS: These findings demonstrate that behavioral and emotional problems tend to increase with age and cluster in the same individuals. The prevalence rates of behavioral problems in Chinese adolescents are comparable to those reported in Western countries. In contrast to findings for Western samples, parent reports of behavioral problems were highly correlated with teacher reports. PMID- 11437023 TI - Psychosocial and risk behavior correlates of youth suicide attempts and suicidal ideation. AB - OBJECTIVE: To identify the independent psychosocial and risk behavior correlates of suicidal ideation and attempts. METHOD: The relationships between suicidal ideation or attempts and family environment, subject characteristics, and various risk behaviors were examined among 1,285 randomly selected children and adolescents, aged 9 through 17 years, of whom 42 (3.3%) had attempted suicide and 67 (5.2%) had expressed suicidal ideation only. The youths and their parents were enumerated and interviewed between December 1991 and July 1992 as part of the NIMH Methods for the Epidemiology of Child and Adolescent Mental Disorders (MECA) Study. RESULTS: Compared with subjects with suicidal ideation only, attempters were significantly more likely to have experienced stressful life events, to have become sexually active, to have smoked more than one cigarette daily, and to have a history of ever having smoked marijuana. After adjusting for sociodemographic characteristics, a statistically significant association was found between suicidal ideation or attempt and stressful life events, poor family environment, parental psychiatric history, low parental monitoring, low instrumental and social competence, sexual activity, marijuana use, recent drunkenness, current smoking, and physical fighting. Even after further adjusting for the presence of a mood, anxiety, or disruptive disorder, a significant association persisted between suicidal ideation or attempts and poor family environment, low parental monitoring, low youth instrumental competence, sexual activity, recent drunkenness, current smoking, and physical fighting. CONCLUSION: Low parental monitoring and risk behaviors (such as smoking, physical fighting, alcohol intoxication, and sexual activity) are independently associated with increased risk of suicidal ideation and attempts, even after adjusting for the presence of psychiatric disorder and sociodemographic variables. PMID- 11437024 TI - Illness-related concerns of mothers of children with congenital heart disease. AB - OBJECTIVE: This study examined concerns expressed by mothers of children with congenital heart disease (CHD). The relationships among mothers' concerns, medical severity, and mother's emotional state were examined at two points in time. METHOD: Thirty-eight mothers of children with CHD aged 3 to 16 completed semistructured interviews and rating scales during hospitalization and 2 to 4 weeks after discharge. Mothers rated their distress about illness-related concerns, as well as their own depressed mood and anxiety. Mothers and two cardiologists rated the medical severity of each child's disease. RESULTS: Mothers' concerns were reliably grouped into five categories: medical prognosis, quality of life, psychosocial functioning, effects on family, and financial issues. During hospitalization, mothers were most concerned about medical prognosis. Distress about most concerns decreased postdischarge, as did mother's anxiety and depressed mood. Mothers' perceptions of medical severity were associated with distress about psychosocial issues postdischarge. Mother's anxiety was not associated with number of concerns reported, or with distress about those concerns. Maternal depressed mood was associated with fewer illness related concerns, but greater distress about those concerns. CONCLUSIONS: Illness related concerns can be meaningfully categorized and are not necessarily a function of disease severity or mother's emotional state. An awareness of common concerns will improve clinical care by enabling practitioners to anticipate and address concerns in a proactive way. The results may inform the development of supportive mental health interventions for families of children with CHD. PMID- 11437025 TI - Genetics of childhood disorders: XXVIII. Autoimmunity, part I. PMID- 11437026 TI - Venous thromboembolism from air travel: the LONFLIT study. AB - The LONFLIT study was planned to evaluate the incidence of deep venous thrombosis (DVT) occurring as a consequence of long flights. In the Lonflit study 355 subjects at low-risk for DVT and 389 at high-risk were studied. Low-risk subjects had no cardiovascular disease and used no drugs. All flights were in economy class. The average flight duration was 12.4 hours (range, 10-15 hr). The mean age of the studied subjects was 46 years (range 20-80 yr, SD 11; 56% males). DVT diagnosis was made by ultrasound scans after the flights (within 24 hours). In low-risk subjects no events were recorded while in high-risk subjects 11 had DVT (2.8%) with 13 thromboses in 11 subjects and 6 superficial thromboses (total of 19 thrombotic events in 389 patients [4.9%]). In the Lonflit2 study the authors studied 833 subjects (randomized into 422 control subjects and 411 using below knee stockings). Mean age was 44.8 years (range, 20-80 yr, SD 12; 57% males). The average flight duration was 12.4 hours. Scans were made before and after the flights. In the control group there were 4.5% of subjects with DVT while only 0.24% of subjects had DVT in the stockings group. The difference was significant. The incidence of DVT observed when subjects were wearing stockings was 18.75 times lower than in controls. Long-haul flights are associated to DVT in some 4 5% of high-risk subjects. Below-knee stockings are beneficial in reducing the incidence of DVT. PMID- 11437027 TI - Differences in frequency of the deletion polymorphism of the angiotensin converting enzyme gene in different ethnic groups. AB - A polymorphism characterized by the insertion or deletion of a 287-bp Alu repeat sequence in intron 16 of the angiotensin-converting enzyme gene determines about half the serum angiotensin-converting enzyme level variability among individuals. The deletion polymorphism is associated with higher levels of angiotensin converting enzyme and perhaps with a greater risk of cardiovascular diseases. The relative frequency of this genetic polymorphism in different ethnic groups is not known. The objective of this study was to compare the frequency of angiotensin converting enzyme gene insertion/deletion polymorphism in different ethnic groups. Angiotensin-converting enzyme genotype was determined in middle-aged healthy hospital workers of three different ethnic origins (African Americans, whites, and Indians). There were 142 African Americans, 136 Indians, and 82 whites. The distribution of the deletion-deletion, insertion-deletion, and insertion-insertion genotypes in African Americans (29%, 60%, and 11%, respectively), Indians (19%, 50%, and 31%, respectively) and whites (29%, 40%, and 31%, respectively) was significantly different (p = < 0.005). The frequency of the deletion allele among African Americans, Indians, and whites (0.59, 0.49, and 0.44, respectively) was also significantly different (p=0.05). African Americans had the highest frequency of deletion allele and the lowest frequency of the insertion-insertion genotype among the three groups. The frequency of the deletion polymorphism of the angiotensin-converting enzyme gene is different among African Americans, whites, and Indians. This may be important in relation to the high risk of cardiovascular morbidity and mortality in African Americans and may be relevant in explaining differences in cardiovascular diseases in different populations. This finding also emphasizes the importance of studying angiotensin-converting enzyme gene polymorphism in genetically homogenous populations. Because of the small size of this study, however, these findings need further confirmation. PMID- 11437028 TI - Improvement of peripheral and cardiopulmonary performance after a short-term exercise program with additive prostaglandin E1. AB - In patients with intermittent claudication, the walking distance can be increased, both by means of several months of intensive training and administration of IV prostaglandin E1 (PGE1) for 4 weeks. The aim of this study was, therefore, to investigate whether the combination of intensive training and PGE1 infusions during pedalergometry can increase peripheral and cardiopulmonary performance after 2 weeks. Ten patients with intermittent claudication received a once-daily intravenous infusion of 60 microg PGE1 over 2 hours during pedalergometry. In addition, a physical training program was carried out mornings and afternoons, as well as progressive treadmill training. Walking distance (3 km/h, 12%) and cardiopulmonary performance were determined at the beginning and end of the 2-week treatment. Results were compared with those of a historical control group having received a similar training program without PGE1. The initial walking distance increased from 71 to 166 m (134%). At the same time, peak work load increased by 108%, and the physical work capacity by 100%. Cardiopulmonary function improvement was reflected in all the parameters investigated (peak VO2; peak VO2/peak work load ratio; slope of deltaVO2/deltat; RER). Compared with the historical control group, the difference between the two groups with regard to the increase in walking distance was significant in favor of the combined training program with PGE1. The combination of short-term intensive training and PGE1 infusions during pedal ergometry significantly improves both the peripheral as well as the highly restricted functional capacity in patients with intermittent claudication. PMID- 11437029 TI - Effects of atrial pacing on QT dispersion in patients with coronary artery disease without angina pectoris and ST segment depression. AB - The aim of this study was to investigate QT dispersion during atrial pacing in patients with coronary artery disease (CAD) without clinical ischemia, such as angina pectoris and ST segment depression. Thirteen patients with normal coronary arteries and 42 patients with CAD (12 with single-vessel, 16 with two-vessel and 14 with three-vessel disease) having no angina pectoris or ST segment depression during atrial pacing with maximum rate of 120/minute were enrolled in the study. Twelve-lead surface ECGs were recorded at 100 mm/second paper speed before pacing, at maximum pacing rate, and during the recovery period for measurement of QT interval parameters. Corrected QTd (QTcd) increased from 43.4 +/- 8.1 to 49.3 +/- 9.5 ms (p < 0.05) in the control group, from 46.1 +/- 8.1 to 74.3 +/- 7.7 ms (p < 0.0001) in the single-vessel disease group, from 48.5 +/- 10.4 to 93.8 +/- 22.1 ms in the two-vessel disease group (p < 0.0001), and from 49.7 +/- 13.6 to 128.5 +/- 31 ms (p < 0.0001) in the three-vessel disease group at peak atrial pacing period. A positive correlation was found between the severity of CAD and QTcd (r = 0.49, p < 0.0001). It was found that pacing-induced QTc dispersion identifies coronary disease extent, even when there is no ST depression or T wave inversion during pacing. PMID- 11437030 TI - Treatment of patients with thromboangiitis obliterans with cyclophosphamide. AB - This report describes a small, nonrandomized trial of cyclophosphamide in the treatment of patients with advanced thromboangiitis obliterans (TAO) with modest results. The rationale of the treatment was based on the immunopathogenesis of the disease, ie, autoimmune vasculitis of peripheral arteries. Twelve male patient volunteers with TAO were included for the trial. Diagnosis was based on the history of chronic smoking or tobacco chewing, clinical features of ischemia of peripheral vessels, radioarteriography showing arterial block, and characteristic histopathologic changes of affected arteries. Cyclophosphamide (400 mg) was given intravenously daily to the patients for 7 days followed by daily oral administration of 100 mg cyclophosphamide for another 7 weeks. Clinical conditions of the patients started to improve during the third week of the treatment and maximum benefit was noticed at the end of the treatment. There was significant decrease of intermittent claudication and twentyfold increase of claudication distance as well as relief of rest pain. Before starting treatment 6 patients had developed ulcers on their affected limbs; these healed completely in 2, partially healed in another 2, and showed no improvement in the remaining 2, who never stopped smoking. However, immunosuppressive therapy failed to show any improvement of arterial block, as evidenced by radioarteriography and any significant increase of skin temperature over the affected limbs. Nevertheless, histopathologic studies of biopsies taken from the diseased arteries after completion of therapy showed decreased influx of lymphocytes and plasma cells in the thrombi as well as in the arterial walls in comparison to the biopsies taken before the start of treatment. During the treatment the degree of immunosuppression was monitored by blood leukocyte and lymphocyte counts, which were kept between 4,000/mm3 and above 3,000/mm3 and not less than 500/mm3, respectively, indicating modest immunosuppression and no serious complications. All patients were followed up for 1 year. Only 2 patients, who resumed smoking, had relapse. PMID- 11437031 TI - Relationship between internal risk factors for development of decubitus ulcers and the blood flow response following pressure load. AB - The objective of this study was to investigate the extent to which internal risk factors for the development of decubitus ulcers are related to the blood flow response following the relief of a pressure load. There were 122 nursing home patients (43 men, 69 women, mean age: 81 +/- 8 years; range: 60-97). The following potential, internal risk factors for the development of decubitus ulcers were assessed: chronic disorders (diabetes mellitus, cardiovascular disease [congestive heart failure, history of myocardial infarct or angina pectoris] and cerebrovascular accident), fever, blood pressure, nutritional status, serum hemoglobin concentration, and serum urea and serum creatinine concentrations. Skin temperature response (latency time and total response time) was measured following relief of a 100 kPa test pressure. The presence of cardiovascular disease, cerebrovascular accident, poor nutritional condition, high serum urea and male gender showed a significant relationship with an impaired blood flow response. The delayed latency found showed a similarity to the so-called "no-reflow phenomenon." The association of cardiovascular disease and a cerebrovascular accident with a delay in the blood flow response may result from endothelial damage. A poor nutritional condition may be associated with a deficit of scavengers of oxygen-derived free radicals. The presence of free radicals may damage endothelium during reperfusion, thus influencing the blood flow response. The association of high serum urea with delayed vasodilatation may theoretically be explained by the association of serum urea and impaired kidney functioning, since the kidney is an important organ in the production of vasoactive substances. Serum urea can also be considered a measure for nutritional condition. Gender may function as a substitute for other, unmeasured factors that are related to blood flow response. PMID- 11437032 TI - ST elevation in the anterior precordial leads during right ventricular infarction: lessons learned during primary coronary angioplasty--a case report. AB - ST elevation in the anterior leads may be due to isolated right ventricular infarction associated with occlusion of a nondominant or codominant right coronary artery. The authors report a case of isolated right ventricular infarction from a dominant right coronary artery's proximal occlusion in the presence of collateral circulation provided by the left coronary artery. Extensive damage occurred owing to compromise of the collateral circulation during primary coronary angioplasty. This is an unusual angiographic pattern for isolated right ventricular infarction. The potential consequences of percutaneous interventions to collateral vessels is discussed. PMID- 11437033 TI - Severe lower limbs lymphedema of late onset revealing polysplenia syndrome--a case report. AB - Polysplenia syndrome includes a group of congenital abnormalities involving the spleen, gastrointestinal tract, liver, and cardiovascular system. A case of severe lower limbs lymphedema occurring in a young woman with polysplenia, azygous continuation of the inferior vena cava, short pancreas, and preduodenal portal vein is reported. Lower limb lymphedema could represent a new clinical manifestation associated with polysplenia syndrome. Lymphedema could be the result of a primary anomaly of the lymphatic system or be induced by high pressure in the venous system or by compression of the lymphatic circulation by the hypertrophic azygous vessels in the posterior mediastinum. PMID- 11437034 TI - Massive right atrial thrombosis due to Hickman catheter requiring open heart surgery--a case report. AB - The risk of catheter-related sepsis and clot formation in patients with an indwelling central venous line is well recognized. However the risk of developing a large thrombus in the right atrium is less well known. Right atrial thrombus can be a life-threatening complication of long-term indwelling catheters and should be considered in the workup of these patients. A case of a 71-year-old lady who developed a large atrial thrombus due to a Hickman catheter is reported that required open heart surgery. Incidence, pathophysiology, clinical presentation, diagnosis, and treatment options are also reviewed. PMID- 11437035 TI - A giant left ventricular thrombus in a patient with acute myocardial infarction- a case report. AB - The authors report a patient with acute anteroseptal myocardial infarction with a giant left ventricular thrombus at the apex. The patient also had nephrotic syndrome due to diabetic nephropathy. Coronary angiography showed 90% stenosis at segment 6 of the left anterior descending coronary artery. Percutaneous transluminal coronary angioplasty and intracoronary stenting were performed on the 30th day, and effective coronary blood flow was obtained. Heparin was injected intravenously for the first 7 days, and warfarin was administered thereafter. The left ventricular thrombus disappeared after 46 days. No evidence of arterial thromboembolism was found during the disappearance of the left ventricular thrombus as determined by echocardiography. PMID- 11437036 TI - The relationship of coronary artery disease and carotid Doppler flow velocity and resistance index in patients with no significant carotid stenosis. PMID- 11437037 TI - Regarding "adult reversible cardiomyopathy with pituitary adrenal insufficiency caused by empty sella". PMID- 11437038 TI - Endoscopic ultrasound-guided drainage of pancreatic pseudocysts or pancreatic abscesses using a therapeutic echo endoscope. AB - STUDY AIMS: The purpose of this study is to evaluate a new drainage technique for pancreatic pseudocysts or pancreatic abscesses entirely guided by endoscopic ultrasound (EUS) and using an interventional echo endoscope with a linear curved array transducer. PATIENTS AND METHODS: Between July 1996 and September 1999, EUS guided drainage of a pancreatic pseudocyst or pancreatic abscess was carried out in 35 patients (26 men, 9 women; mean age 56.7, range 29-69). The mean size of the 35 pancreatic cysts was 7.8 cm (4-12 cm). Pancreatic pseudocysts were located in the head of the pancreas in two cases, in the body in six cases and in the tail in seven cases. On the other hand, the pancreatic abscesses were located in the tail of the pancreas in 17 cases and in the gastric wall in three cases. The EUS instrument used was the FG 38X endoscope manufactured by Pentax-Hitachi. RESULTS: No major complication occurred except in one case of a pneumoperitoneum, which was managed medically. Placement of the 7-F nasocystic drain was successful in 18/20 cases of pancreatic abscess. Surgery was performed in the two other patients. Concerning the pancreatic pseudocysts, placement of an 8.5-French stent was successful in 10 patients and of a nasopancreatic drain in five patients. In one case, only a puncture-aspiration was performed. One recurrence among the 15 pancreatic pseudocysts and two relapses of the 18 pancreatic abscesses have been observed, over a mean follow-up of 27 months (6-48 months). EUS-guided drainage was successful in 31/35 patients (88.5%); only four patients with pancreatic abscesses underwent surgery. No bleeding occurred during the time of this study. CONCLUSION: Internal drainage of pancreatic pseudocysts and abscesses exclusively performed with an echo endoscope is a safe and efficient method which should be evaluated further in larger studies. PMID- 11437039 TI - Clinical impact of endoscopic ultrasound-guided fine needle aspiration biopsy in patients with upper gastrointestinal tract malignancies. A prospective study. AB - BACKGROUND AND STUDY AIMS: Several studies have evaluated the accuracy of endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS-FNAB) in the upper gastrointestinal tract, but so far no studies have specifically evaluated the clinical impact of EUS-FNAB in upper gastrointestinal tract cancer patients. In this consecutive and prospective study, EUS-FNAB was only performed if a positive malignant finding would change the therapeutic strategy. PATIENTS AND METHODS: Between 1997 and 1999, 307 consecutive patients were referred for EUS with a diagnosis or strong suspicion of esophageal, gastric or pancreatic cancer; 274 patients were potential candidates for surgical treatment and had EUS. According to predefined impact criteria, 27% (75/274) of the patients had EUS FNAB for staging or diagnostic reasons. RESULTS: The overall clinical impact of EUS-FNAB was 13%, 14%, and 30% in esophageal, gastric, and pancreatic cancer, respectively. The staging-related clinical impact was similar for all three types of cancer (11-12.5%), whereas the diagnosis-related impact was highest in pancreatic cancer patients (86%). EUS-FNAB was inadequate in 13% and gave false negative results in 5%. The overall sensitivity, specificity and accuracy for EUS FNAB were 80%, 78% and 80%, respectively. No complications related to the biopsy procedure were seen. CONCLUSIONS: If EUS-FNAB was performed only in cases where a positive malignant result would change patient management, then approximately one out of four patients with upper gastrointestinal tract cancer would require a biopsy. With this approach the actual clinical impact of EUS-FNAB ranged from 13% in esophageal cancer to 30% in pancreatic cancer. EUS-FNAB plays a limited, but very important clinical role in the assessment of upper gastrointestinal tract cancer. PMID- 11437040 TI - Endoscopic ultrasound-guided fine-needle aspiration in focal pancreatic lesions: a prospective intraindividual comparison of two needle assemblies. AB - BACKGROUND AND STUDY AIMS: The results of endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) in focal pancreatic lesions are less impressive than those in the mediastinum. The aim of this prospectively randomized study was to compare two commercially available needle assemblies with regard to handling and cytopathological yield. PATIENTS AND METHODS: A total of 30 patients (19 men, 11 women; mean age 61) with focal pancreatic lesions underwent EUS-FNA with each of the two needles (GIP, Wilson-Cook). The sequence was randomized for the examiner and blinded for the cytologist. Three patients had to be excluded because of the impossibility of sample assignment or patient follow-up. EUS-FNA was performed using the standard technique with linear echo endoscopes. RESULTS: None of the characteristics evaluated by the examiner differed significantly between either of the needles. Inadequate results were obtained in 11% using the GIP needle, but in none with the Wilson-Cook needle. GIP needle cytology revealed malignancy in 11 patients (sensitivity, specificity, and accuracy were 55%, 100%, and 65%, respectively, including inadequate results). The aspirates obtained with the Wilson-Cook needle identified malignancy in 16 patients (sensitivity, specificity, and accuracy were 85%, 100%, and 89%, respectively). CONCLUSIONS: No statistically significant differences were detected in the handling of either of the two needle assemblies. No complications were reported using the GIP needles. However, in four procedures breakages of the outer Teflon sheath of the Wilson Cook needle occurred, and in another four cases re-insertion of the stylet was impossible. Nevertheless, cytopathologic results were significantly better with the Wilson-Cook needle. PMID- 11437041 TI - Ofloxacin and ursodeoxycholic acid versus ursodeoxycholic acid alone to prevent occlusion of biliary stents: a prospective, randomized trial. AB - BACKGROUND AND STUDY AIMS: Biliary plastic stents are highly effective in the treatment of malignant biliary obstruction, but may become occluded over time, leading to jaundice and cholangitis. Stent occlusion is thought to be caused by bacterial adhesion and formation of biofilm. This study was carried out to assess whether treatment with ofloxacin in combination with ursodeoxycholic acid is superior to ursodeoxycholic acid alone in preventing stent occlusion. PATIENTS AND METHODS: Patients with obstructive jaundice due to inoperable malignant disease underwent placement of a straight 11.5-Fr polyethylene stent. After stent insertion, the patients were randomly assigned to receive either ofloxacin (200 mg b.i.d.) with ursodeoxycholic acid (250 mg t.i.d.) or ursodeoxycholic acid alone. The end points of the study were the frequency of stent occlusions, the time to stent occlusion, and the safety of the two regimens. RESULTS: Fifty-two patients were enrolled, of whom 26 were assigned to the combined therapy group and 26 to the control group. Thirty patients were suffering from pancreatic cancer, 13 from gallbladder or bile duct cancer, and nine had metastases from other malignant tumors. Eight stent occlusions (31%) occurred in the ofloxacin group and six (23 %) in the control group (P = 0.76). The mean times to stent occlusion were 95 +/- 9 days and 101 +/- 9 days, respectively (P = 0.91). No significant differences regarding survival time or safety were observed between the two groups. CONCLUSIONS: Ofloxacin in combination with ursodeoxycholic acid is not superior to ursodeoxycholic acid alone in preventing stent occlusion in patients with malignant obstructive jaundice. PMID- 11437042 TI - The clinical significance of common bile-duct dilatation in patients without biliary symptoms or causative lesions on ultrasonography. AB - BACKGROUND AND STUDY AIMS: Although abdominal ultrasonography (US) is a good initial screening method for detection of biliary tract disease, we sometimes encounter patients who only have findings of dilatation of the common bile duct (CBD) on US, without specific biliary symptoms or jaundice. This study aimed to evaluate the causes and clinical significance of dilatation of the CBD in patients without biliary symptoms, jaundice, or causative lesions at US. PATIENTS AND METHODS: A total of 77 patients who had no biliary symptoms and whose internal CBD diameter was more than 7 mm, without definite causative lesions on US, were enrolled. Of these, 49 underwent endoscopic retrograde cholangiopancreatography (ERCP) and 28 underwent follow-up US or computed tomography (CT) instead of ERCP. We excluded patients whose bilirubin level had increased beyond the upper normal level or who had previous history of upper abdominal surgery including cholecystectomy. RESULTS: The ERCP findings were as follows: no lesion in 20 patients (40.8%), juxtapapillary duodenal diverticulum (JDD) in 11 (22.5%), benign stricture in ten (20.4%), distal CBD mass in two (4.1%), choledochal cyst in two (4.1%), anomalous union of the pancreaticobiliary duct (AUPBD) in two (4.1%), and choledochal cyst with AUPBD in two (4.1%). There were no differences in age or in alkaline phosphatase or gamma-glutamyl transpeptidase (GGT) levels between the patients who had causative lesions revealed at ERCP and those who did not. Among the 28 patients who did not undergo ERCP, 12 had returned to normal and eight had no change in CBD diameter on follow up US. Among eight patients who underwent CT, there were four with normal findings, one with JDD, and three with suspected choledochal cysts. CONCLUSIONS: We detected a significant number of causative biliary tract lesions in asymptomatic adults with dilatation of the CBD on routine abdominal US; no laboratory or demographic parameters were useful for discrimination. Further diagnostic study will be helpful for the early diagnosis of biliary tract disease in such patients. PMID- 11437043 TI - Risk factors suggesting malignant transformation of gastric adenoma: univariate and multivariate analysis. AB - BACKGROUND AND STUDY AIMS: Since gastric adenomas are precancerous lesions, polypectomy is necessary. However, there have been no reports suggesting factors capable of predicting malignant transformation of gastric adenomas removed by endoscopic snare polypectomy (ESP) or endoscopic mucosal resection (EMR) in Korea, a country in which gastric cancer is a major problem. The aim of this paper was to elucidate the risk factors suggesting malignant transformation of gastric adenomas removed by ESP or EMR at our center. PATIENTS AND METHODS: Between November 1994 and June 1999, 118 gastric adenomas diagnosed on the basis of endoscopy and histological examinations of the forceps biopsy specimens obtained were treated by ESP or EMR at our department. Factors capable of predicting malignancy were searched for in the endoscopy reports, still photographs, and histopathological findings. RESULTS: Eight of the 118 adenomas ultimately proved to have malignant foci. In the univariate analysis, four of the variables studied--location, histological type, surface redness, and degree of dysplasia--had a statistically significant relationship with malignant transformation. In the multivariate analysis, only the degree of dysplasia had a statistically significant relationship with malignant transformation. CONCLUSIONS: These results suggest that a diagnosis of high-grade dysplasia in forceps biopsy material should be considered an absolute indication for ESP or EMR. PMID- 11437044 TI - Metastatic tumors to the stomach: analysis of 54 patients diagnosed at endoscopy and 347 autopsy cases. AB - BACKGROUND AND STUDY AIMS: There have been several published reports on metastatic lesions in the stomach, but the numbers of cases have been limited due to the low frequency of the condition. The present study examined the clinicopathological features of metastatic tumors in the stomach from distant sites in a large series of cases. PATIENTS AND METHODS: A total of 389 patients with gastric metastases from solid malignant tumors were examined between 1968 and 1998 at our institution. Of these, 347 were identified from a series of 6380 autopsy cases; 54 patients were diagnosed endoscopically while alive, 12 of whom had confirmation of the condition at autopsy. RESULTS: In the endoscopically diagnosed cases, the metastases presented as solitary (65%) or multiple lesions (35 %), and were more frequently located in the middle or upper third of the stomach. Although the endoscopic appearance often resembled that of submucosal tumor (51%) or primary gastric cancer (39%), the final diagnosis was easily obtained in over 90% of cases from endoscopic biopsies. In two cases of lung cancer and breast cancer, gastric metastases were found before the primary tumors. In the autopsy cases with solid malignancies, metastatic lesions to the stomach were found in 5.4%, and the lung, breast, and esophagus were common primary sites. Malignant melanoma was the most frequent tumor to metastasize to the stomach (29.6%). CONCLUSIONS: Since metastatic lesions to the stomach are rare, the above characteristics of the lesions should be borne in mind, and biopsies should be taken for precise diagnosis during endoscopic examinations. PMID- 11437045 TI - The impact of endoscopists' experience and learning curves and interendoscopist variation on colonoscopy completion rates. AB - BACKGROUND AND STUDY AIMS: Since its introduction in the late 1960s, the technology of colonoscopy has developed rapidly and the competence of endoscopists has increased. Nevertheless, it is not always possible to perform a complete colonoscopy. The aim of this study was to assess, in a population-based setting, the endoscopist-related factors influencing the completion rate, taking into account patient characteristics and changes in the technology over time. PATIENTS AND METHODS: All colonoscopy records between 1979 and 1995 in one Swedish county (population 258,000) were retrieved. Information was obtained about patient demographics, date of examination, endoscopist, indications, presence of diverticulosis, type of colonoscopy, findings, level of completion, complications, and reasons for incomplete colonoscopy. Completion rates were examined by univariate and multivariate analyses. RESULTS: Of 5,494 colonoscopies, 4,153 (75%) were complete, and 78% were diagnostic and 22% therapeutic. In 70 %, findings were pathological, and 30 % were normal. The overall 30-day endoscopist-related complication rate was 0.3 %. Over time, the proportion of colonoscopies performed by endoscopists with greater experience increased and so did the completion rates. Completion rates were influenced by endoscopist's experience and to some extent by the intensity (the number of colonoscopies performed by the endoscopist during the previous 90 days). There was a large interendoscopist variation, at each level of experience, in the ability to perform a complete colonoscopy, implying substantial differences between individual learning curves. CONCLUSIONS: The completion rate has increased over time, the major reason being greater competence of endoscopists. The finding of large interendoscopist variation, at each level of experience, in the ability to perform a complete colonoscopy supports the monitoring of endoscopists to maintain and improve performance. PMID- 11437046 TI - Endoscopic removal of sharp foreign bodies impacted in the esophagus. AB - BACKGROUND AND STUDY AIMS: Impacted sharp foreign bodies in the esophagus can be very difficult to manage. When attempts are made to remove such objects inappropriately, life-threatening complications such as perforation can occur. The aim of this study was to evaluate the safety and efficacy of endoscopic removal of impacted sharp foreign bodies in the esophagus using proximal dilatation with an oral side balloon or transparent cap. PATIENTS AND METHODS: A total of 22 patients (10 men, 12 women) with impacted sharp foreign bodies in the esophagus underwent endoscopic extraction. The following technique was successfully performed at our hospital. An oral side balloon (Top Co., Japan) for esophageal variceal sclerotherapy was attached to the distal part of the endoscope. With the patient under local anesthesia, the endoscope was inserted as far as the proximal part of the esophageal foreign body. The oral side balloon was then gradually inflated. Dilatation of the proximal part of the esophagus made it possible to release the impacted sharp foreign body from the esophageal wall. A transparent cap was used for foreign bodies in the upper esophagus when there were difficulties with the oral side balloon. RESULTS: The types of foreign body removed were fish bones (n = 9), press-through packages (n = 8), chicken bones (n =3), dentures (n = 1), and a wrist watch (n = 1). Endoscopic removal was successful in all but one of the cases, in which a fish bone had to be extracted surgically. CONCLUSIONS: The proximal dilatation method using an oral side balloon or transparent cap is safe and effective in removing sharp foreign bodies from the esophagus, avoiding surgery and possible perforation. PMID- 11437047 TI - Freely accessible endoscope channels improve efficacy of cleaning. AB - BACKGROUND AND STUDY AIMS: Inadequate cleaning and disinfection of medical devices, including flexible endoscopes, can result in the transmission of micro organisms to patients. The aim of this study was to investigate the influence of the design of medical devices on the efficacy of manual cleaning of endoscope channels. MATERIALS AND METHODS: The investigation was carried out using four endoscopes (two duodenoscopes and two gastroscopes). The air/water channels of one duodenoscope and one gastroscope were freely accessible and could be brushed. The instrumentation and the air/water channels were contaminated with blood containing Enterococcus faecium as a test organism. After manual cleaning of the channels by flushing and, where possible, brushing, the recovery rates for the test organism were studied. RESULTS: The comparable rates for recovery of the test organism after cleaning of the instrumentation channels proved that the method used was reproducible. With regard to the air/water channels, the rate of micro-organisms in the cleaning solution recovered after flushing alone was a maximum of 3 % relative to the rate detected after brushing and flushing. CONCLUSIONS: The data collected in the study show that only flushing channels that are not freely accessible resulted in significantly lower (P<0.001) recovery rates for the test organism. In practice, this means that contamination may remain in the channels, and it shows that the design of a medical device has an important influence on the reprocessing of reusable instruments such as flexible endoscopes. PMID- 11437048 TI - Determination of glutaraldehyde residues on flexible endoscopes after chemothermal treatment in an endoscope washer-disinfector. AB - BACKGROUND AND STUDY AIMS: Although there are several cases of glutaraldehyde induced colitis following colonoscopy there is only one study on residues after disinfecting. Our report describes the determination of water-soluble glutaraldehyde residues after processing endoscopes with a glutaraldehyde containing disinfectant. MATERIALS AND METHODS: A gastroscope and a colonoscope were processed in a washer-disinfector and then immersed in 201 of NaCl solution. Glutaraldehyde was determined by high performance liquid chromatography in 84 samples. Samples were taken after immersing the endoscopes for 5 h and 24 h with and without rinsing the channels. RESULTS: After 5 h and with the channels having been rinsed, the median for colonoscopes was only 0.409 mg/l. The highest level of glutaraldehyde was 0.846 mg/l. CONCLUSIONS: After endoscopes have been processed in the washer-disinfector there is no risk of a glutaraldehyde-induced colitis, proctitis or diarrhoea. PMID- 11437049 TI - Microwave recanalization: thermal effects with nitinol and stainless-steel stents. AB - BACKGROUND AND STUDY AIMS: A previous study we conducted evaluated the thermal effects of microwave irradiation when attempting recanalization of a nitinol stent occluded by tumor ingrowth. The present study evaluates the thermal effects of microwave irradiation on different stents in vitro. METHODS: The thermal response to microwave energy was evaluated in a muscle-equivalent phantom containing a nitinol (Accuflex) or stainless-steel stent (Wallstent). The phantom temperature was monitored continuously at different distances from an electrode, with either stent type in place. The tip of the electrode was either in contact with the stent or not in contact with it. RESULTS: The steel stent showed a smaller temperature increase than nitinol at 2 mm from the electrode. Without electrode contact, the stents did not differ significantly during the first 40s of irradiation. The stents did not differ significantly with regard to temperatures further from the electrode. When the electrode was in contact with the stent, the rise in temperature was significantly greater with the stainless steel stent than with the nitinol one, but the phantom temperature did not exceed 45 degrees C with either of the stents. CONCLUSIONS: In patients with a Wallstent stainless-steel stent, microwave coagulation therapy to recanalize stent occlusions appears to be feasible in the same conditions as with an Accuflex nitinol stent. PMID- 11437050 TI - Endoscopic ultrasonography (EUS) and EUS-fine-needle procedures: fascinating technique but little clinical impact? PMID- 11437051 TI - Training simulators and education in gastrointestinal endoscopy: current status and perspectives in 2001. PMID- 11437052 TI - Esophageal involvement in Stevens-Johnson syndrome. AB - The authors describe the endoscopic aspect of esophageal lesions in five children with Stevens-Johnson syndrome. Lesions involve the entire esophagus, with blistering of the epithelium leading to large ulcerations of the mucosae. Esophageal involvement is probably underestimated in Stevens-Johnson syndrome and may worsen dysphagia caused by oral lesions, leading to malnutrition. Enteral nutrition can be helpful to provide feeding, limit weight loss, and support skin healing. No strictures were diagnosed during the follow-up period of these patients. PMID- 11437053 TI - Laparoscopic-assisted appendicectomy (LAA): a novel advance on an established procedure. PMID- 11437054 TI - Collagenous duodenitis and collagenous colitis: a short clinical course as evidenced by sequential endoscopic and histologic findings. PMID- 11437055 TI - Endoscopic closure of esophagothoracocutaneous fistula with collagen plug and fibrin glue. PMID- 11437056 TI - Intestinal tuberculosis after successful treatment of advanced high-grade non Hodgkin's lymphoma and AIDS. PMID- 11437057 TI - Agricultural exposure history among African-American farmers in Georgia. AB - Agricultural exposures differ across the United States by region, calendar time period, and agricultural practice, but most of the published literature focuses on white men in the Midwest. A pilot study was conducted to explore the breadth and diversity of farming practices over time among African-American farmers in Georgia whose exposures may differ in important ways. Using a comprehensive life events calendar questionnaire, 17 male African-American farmers aged 36 to 86 yr residing in southeastern Georgia were interviewed regarding their agricultural history in July 1997. Most men (15/17) reported working on multiple farms in their lifetime; 3 men worked on 5 different farms during their lifetime. These farmers reported using more chemicals during their lifetime than farmers in the Midwest. Used motor oil was the most frequently reported insecticide applied to animals; this apparently common practice has not been described in the literature and should be better understood since its use may result in dermal exposure to polyaromatic hydrocarbons. Better characterization of regionally specific farming history and individual farming practices will facilitate studies of the health effects of farming. PMID- 11437058 TI - Combined exposure to DEET (N,N-diethyl-m-toluamide) and permethrin-induced release of rat brain mitochondrial cytochrome c. AB - The release of cytochrome c from the mitochondrial intermembrane space can induce apoptosis. The levels of mitochondrial cytochrome c in rat brain following a single dermal dose of 400 mg/kg of DEET, and of 1.3 mg/kg of permethrin, alone or in combination were determined. Rats were sacrificed at a time interval of 0.5, 1, 2, 4, 8, 16, 24, 48, or 72 h after dosing. Brain mitochondria were isolated and the levels of cytochrome c were measured using reversed-phase high performance liquid chromatography (HPLC) with ultraviolet (UV) detection. Average percentage recovery of cytochrome c spiked with control rat brain mitochondria was 83.2 +/- 8.9%. Limits of detection and quantitation were 1 and 5 ng, respectively. The results showed that a single dermal dose of a combination of DEET and permethrin significantly increased the release of brain mitochondrial cytochrome c starting 24 h after treatment. DEET and permethrin alone did not affect the release of cytochrome c. The results indicate that combined exposure to DEET and permethrin might induce the apoptotic processes in rat brain as seen by the release of cytochrome c. PMID- 11437059 TI - Glucose feeding exacerbates parathion-induced neurotoxicity. AB - Excessive dietary intake of sugars could alter various biotransformation processes and the pharmacological and toxicological properties of numerous xenobiotics. In the present study, the effects of glucose supplementation were examined on the neurotoxicity of the organophosphorus (OP) pesticide parathion (PS) and its active metabolite, paraoxon (PO), a potent inhibitor of acetylcholinesterase (AChE). Rats (n = 6-12/treatment group) were given free access to tap water or 15% glucose (w/v) in tap water beginning 7 d prior to OP toxicant exposure. Food, caloric intake, and body weight were measured daily. Animals were challenged with either PS (4.5, 9, or 18 mg/kg, sc) or PO (0.3 0.5, or 0.7 mg/kg, sc) and clinical signs of neurotoxicity (i.e., autonomic dysfunction, involuntary movements) were recorded daily for the following 13 d. Glucose feeding was associated with a dramatic drop (approximately 50%) in feed intake and an increase (approximately 20% in total caloric consumption over the 7 d prior to OP exposure. Functional toxicity associated with PS exposure was increased in glucose-fed (GF) rats, but the glucose diet had no apparent effect on clinical signs of toxicity following PO treatment. Glucose feeding increased the magnitude of AChE inhibition in the frontal cortex and plasma at lower dosages (i.e., 4.5 and 9 mg/kg) 3 d following PS treatment. Time-course studies (3, 7, and 11 d after PS exposure, 18 mg/kg, sc) indicated significantly greater brain and plasma AChE inhibition in glucose-fed animals at later time points. In contrast, glucose feeding had no effect on the degree of AChE inhibition following PO exposure. Neither liver microsomal oxidative desulfuration of PS, nor liver or plasma paraoxonase, nor liver or plasma carboxylesterase activities were measurably affected by glucose feeding. Downregulation of muscarinic receptors 7 d after PS exposure (18 mg/kg, sc) was more extensive in GF rats. It is postulated that excessiveglucose consumption decreases the intake of other dietary components, in particular amino acids, limiting the de novo synthesis of AChE and consequent recovery of synaptic transmission. Due to the shorter duration of inhibition following PO exposure, sponta neous reactivation of AChE may be more important than de novo protein synthesis in recovery of function, and thus with the effects of glucose feeding on its toxicity. Individuals that derive a large proportion of their calories from sugars may be at higher risk of acute toxicity from organophosphorus pesticides such as PS. PMID- 11437060 TI - Organotins disrupt components of glutamate homeostasis in rat astrocyte cultures. AB - A rat cortical astrocyte preparation was used to investigate the effects of organotins on glutamate regulation by astrocytes. Exposure of astrocytes to low levels of organotins produced significant changes in two key components of glutamate homeostasis: glutamine synthetase (CS) activity and the high-affinity transport of L-glutamate. Trimethyltin (TMT), triethyltin (TET), and triphenyltin (TPT) exhibited differential abilities to reduce GS activity and glutamate uptake. Cultures incubated with 1 microM TET or TPT, but not TMT, exhibited a marked decrease in GS activity. Exposure to TET or TPT also produced a significant decrease in glutamate transport activity that was not observed with TMT. These declines in activity were not attributable to cell loss as measured by MTT reduction and lactate dehydrogenase (LDH) leakage. Since the loss of GS activity and transporter activity was not seen with acute organotin exposure, it is most likely attributable to a decreased presence of fully functioning protein. While the attenuation of GS and glutamate transporter activities by organotins does not match their pattern of neurotoxicity, the results indicate the potential for subtoxic concentrations of these compounds to increase extracellular glutamate and interact with other excitotoxic episodes. PMID- 11437061 TI - Effects of oxidative stress on the erythrocyte Na+,K+ ATPase activity in female hyperthyroid patients. AB - This study was planned to determine the effects of free-radical-induced damage on the Na+,K+-ATPase activity of erythrocytes during hyperthyroidism and 4 wk after propylthiouracil (PTU) therapy (400 mg/d). The levels of plasma thiobarbituric acid-reactive substances (TBARS) as a marker of lipid peroxidation, erythrocyte glutathione (GSH) concentration as an antioxidant, blood ATP concentration, and erythrocyte membrane Na+,K+-ATPase activity were determined in female hyperthyroid patients (n = 22, mean age 40.5 +/- 6.5 yr). Before the PTU therapy, plasma TBARS concentration was significantly higher and the levels of blood ATP and erythrocyte GSH and the activity of membrane Na+,K-+-ATPase were significantly lower in the hyperthyroid patients (n = 15 women, mean age 40.8 +/- 7.3 yr). Four weeks after PTU therapy, plasma TBARS concentration was decreased, and levels of erythrocyte GSH and blood ATP and of Na+,K+-ATPase activity of erythrocytes were elevated in the treated patients. There was a significant positive correlation between blood ATP concentration and Na+,K+-ATPase activity, and a negative correlation between plasma TBARS concentration and Na+,K+-ATPase activity before PTU. Our results might help to clarify the effects of the oxidative mechanisms on the erythrocyte membrane Na+,K+-ATPase activity in hyperthyroid patients. PMID- 11437062 TI - Oral treatment of Fischer 344 rats with weathered crude oil and a dispersant influences intestinal metabolism and microbiota. AB - When oil is spilled into aquatic systems, chemical dispersants frequently are applied to enhance emulsification and biological availability. In this study, a mammalian model system was used to determine the effect of Bonnie Light Nigerian crude oil, weathered for 2 d with continuous spraying and recirculation, and a widely used dispersant, Corexit (Cx) 9527, on intestinal microbial metabolism and associated populations. To determine the subchronic dose, concentrated or diluted (1:2, 1:5, 1:10, 1:20) Cx9527 or oil was administered by gavage to Fischer 344 rats and the effect on body weight was determined. Next, rats were treated for 5 wk with oil, dispersant, or dispersant + oil. Body and tissue weights, urine mutagenicity, and the impact on the intestinal microflora and three microbial intestinal enzymes linked to bioactivation were determined in the small and large intestines and cecum. Two tested dispersants, Cx9527 and Cx9500, were toxic in vitro (1:1,000 dilution), and oil was not mutagenic in strains TA98 and TA100(+/ S9). None of the treated rats produced urine mutagens detected by TA98 or TA100. Undiluted dispersant was lethal to rats, and weight changes were observed depending on the dilution, whereas oil generally was not toxic. In the 5-wk study, body and tissue weights were unaffected at the doses administered. Small intestinal levels of azoreductase (AR), beta-glucuronidase (BG), and nitroreductase (NR) were considerably lower than cecal and large-intestinal activities at the same time point. A temporal increase in AR activity was observed in control animals in the 3 tissues examined, and large-intestinal BG activity was elevated in 3-wk controls. No significant changes in cecal BG activity were observed. Oil- or dispersant-treated rats had mixed results with reduced activity at 3 wk and elevated activity at 5 wk compared to controls. However, when the dispersant was combined with oil at 3 wk, a reduction in activity was observed that was similar to that of dispersant alone. One-week nitroreductase activity in the small intestine and cecum was unaffected in the three treatment groups, but elevated activity was observed in the large intestines of animals treated with oil or dispersant. The effect of the combination dose was not significantly different from the control value. Due to experimental error, no 3- or 5-wk NR data were available. By 5 wk of treatment, enterobacteria and enterococci were eliminated from ceca of oil-treated rats. When oil was administered in combination with dispersant, an apparent protective effect was observed on the enterococci and lactose-fermenting and nonfermenting enterobacteria. A more detailed analysis at the species level revealed qualitative differences dependent on the treatment. This study suggests that prolonged exposure of mammals to oil, dispersant, or in combination impacts intestinal metabolism, which ultimately could lead to altered detoxification of oil constituents and coexposed toxicants. PMID- 11437063 TI - Knowledge and use of folic acid supplementation: a study of Colorado women whose pregnancies were affected by a fetal neural tube defect. AB - OBJECTIVES: To determine whether women who have ha da pregnancy in which the fetus is affected by a neural tube defect (NTD) know of current folic acid recommendations; whether the recommendations are followed before or during the pregnancy associated with an NTD and subsequent pregnancies; and to insure that women who have had an NTD-affected pregnancy have access to proper information about NTDs and folic acid recommendations. METHODS: Women living in Colorado who had a pregnancy in 1996 or 1997 in which the fetus had an NTD were interviewed in 1998 about their knowledge and use of folic acid supplementation. RESULTS: Twenty one of 42 eligible women were interviewed. All women first learned of the folic acid recommendations either during or after their affected pregnancy. Only 23.8% of the women took vitamins containing folic acid during the 1 to 3 months before becoming pregnant. None who had a subsequent pregnancy followed the recommendation to consume 4.0 mg/d of folic acid, beginning at least 1 month before conception. Women who had subsequent pregnancies became pregnant on average of 9 months after their affected pregnancy ended. CONCLUSIONS: Most women who have an NTD-affected pregnancy are unaware of the national folic acid recommendations and do not follow these recommendations for subsequent pregnancies. However, such women are receptive to information about folic acid supplementation. Health care providers and public health officials should consider their role in assuring that education is provided in an effective and timely manner to women with NTD-affected pregnancies. PMID- 11437064 TI - Neurodevelopment of adopted children exposed in utero to cocaine: the Toronto Adoption Study. AB - BACKGROUND: Published studies of children's neurodevelopment after in utero exposure to cocaine have not separated intrauterine from postnatal environmental effects as cocaine-using mothers cluster in low socioeconomic classes and have other risk factors. METHODS: To overcome this limitation, a study was done to assess physical and neurodevelopmental characteristics of 52 children: 26 were adopted by parents who sought counselling in the Motherisk Program at the University of Toronto for prenatal cocaine exposure, and 26 were controls matched for maternal intelligence quotient (IQ), socioeconomic status and gestational age. MAIN OUTCOME MEASURES: Head circumference, McCarthy General Cognitive Index (GCI) score, language performance and temperament tests. RESULTS: The children in the study group had smaller head circumferences (34th versus 54th percentiles p = 0.009), lower McCarthy GCI scores (102.8 versus 114.2, p = 0.02), poorer receptive and expressive language performance on the Reynell test, and higher activity levels, less persistence and increased distractibility on temperament tests. On multivariate analysis, cocaine exposure was significantly (p = 0.001) associated with lower IQ and poorer language development independent of intrauterine growth retardation and other potential confounders. INTERPRETATION: By controlling for postnatal environmental factors, this adoption study documents intrauterine developmental risks associated with cocaine exposure. Follow-up into school years is warranted to evaluate the extent of these effects. PMID- 11437065 TI - Epigenetic inheritance associated with human chromosome 14. AB - Within the last decade, there has been sufficient evidence to support the association of epigenetic inheritance or genomic imprinting on human chromosome 14. This has been achieved with studies of imprinting on both human chromosome 14 and mouse chromosome 12, which has the largest homology to human chromosome 14. Initial studies with mouse chromosome 12 aberrations suggested that specific phenotypes due to genomic imprinting were confined to the maternal or paternal mouse chromosome 12, depending on the parent from which the chromosome was inherited. Such findings were later supplemented with human chromosome 14 aberrations that had provided evidence for imprinted intervals on this chromosome, and possibly 2 syndromes associated with a maternal or paternal uniparental disomy affecting chromosome 14. The recent discovery of 2 imprinted genes on human chromosome 14 and mouse chromosome 12 has confirmed genomic imprinting on these chromosomes. These findings will refine our understanding of the clinical consequences of human chromosome 14 aberrations and of the causes of the disease phenotype associated with defective imprinted genes. This article reviews evidence in mice and humans, and the clinical implications for genomic imprinting associated with human chromosome 14. PMID- 11437066 TI - BRCA1, BRCA2 and breast cancer: a concise clinical review. AB - Less than 5% of breast cancers are hereditary, but over 90% of hereditary breast cancers are caused by a mutation of either BRCA1 or BRCA2. The mutation may be inherited from either the maternal or the paternal side of the family. Clinicians should consider specific criteria in the family history to determine when a patient may benefit from counselling and appropriate testing. Testing is generally offered only to patients who are at high risk and is currently estimated to have a sensitivity of about 85%. Test protocols are primarily oriented to detecting frameshift and nonsense mutations that cause premature protein truncations. Missense mutations also occur, but they are less common and sometimes not clearly of clinical significance. Laboratory results need to be correlated with the clinical picture, and genetic counselling is a critical component in maximizing the benefits of testing. In the future, application of more refined clinical criteria, as well as expected improvements in laboratory techniques, will undoubtedly lead to significantly better outcomes and options in surveillance and management for hereditary breast and ovarian cancer syndromes caused by mutations of BRCA1 and BRCA2. PMID- 11437067 TI - Recruiting researchers. PMID- 11437068 TI - Recruiting researchers. PMID- 11437069 TI - The isolation and function of porcine islets from market weight pigs. AB - The efficacy of clinical islet transplantation has been demonstrated with autografts, and although islet allografts have established insulin independence in a small number of IDDM patients, the treatment is confounded by the necessity of immunosuppression. the lack of donor tissue, and recurring islet immunogenicity. These limitations underscore a need to develop therapies to serve the large population of diabetic patients. Porcine islet xenotransplantation, together with a successful immune intervention strategy, may provide the necessary clinical alternative. However, a major obstacle in evaluating this approach has been the difficulty of obtaining adequate volumes of functional islet tissue from pigs. Donors of market weight are preferable to retired breeders due to their abundance, lower animal and husbandry costs. and are more suitable to meet regulatory guidelines for donor tissue for xenotransplantation. We describe a simple isolation procedure that following purification yields a mean of 350,000 IE, corresponding to 179 units of insulin and 1.8 mg of DNA with an islet purity and viability in excess of 85% (n = 317 isolations). In both short- and long-term cell cultures, porcine islets demonstrated glucose responsive insulin secretion. However, this secretion is density dependent, which may have significant consequences in the development of immunoisolation technologies to support porcine islet xenotransplantation. Following implantation into diabetic nude mice, porcine islets remained functional in excess of 1 year. Implantation of a bioartificial pancreas containing porcine islets into pancreatectomized dogs provided significant clinical benefit with an improved diabetic condition. Finally, secretagogue-induced insulin release was demonstrated in vitro from these devices after removal from immunocompetent recipients. Immunohistochemical staining identified well-granulated islets following long-term implantation in both the rodent and canine models. This study demonstrates the ability to isolate porcine islets in clinically relevant numbers from market animals, which survive and remain functional for prolonged periods of time in an immune-deficient or immunoprotected environment. PMID- 11437070 TI - Islet cryopreservation: improved recovery following taurine pretreatment. AB - Simple and efficient freezing methods with maximal postthawing recovery form the basis of ideal cryopreservation. Taurine (2-amino ethanesulfonic acid), an end product of sulphur amino acid metabolism, is one of the most abundant free amino acids in the body. The membrane stabilizing, free radical scavenging, and osmoregulatory roles of taurine have been well documented. We studied the effect of physiological and supra-physiological concentrations (0.3 and 3.0 mM) of taurine on islet cryopreservation. Islet viability on cryopreservation was significantly improved in both the taurine-treated groups (91.9 +/- 2.3% in 0.3 mM and 94.6 +/- 1.58% in 3.0 mM group, p < 0.05) compared with the controls (85.7 +/- 3.4%). Loss of peripheral islet cells was highly reduced in the taurine group, as examined under phase contrast and quantified by islet morphometric analysis (p < 0.05) using a digital image analysis system. Taurine-treated islets showed significant reduction in lipid peroxidation (0.905 and 0.848 nM MDA/microg protein for 0.3 and 3.0 mM taurine, respectively, p < 0.05) compared with control (1.307 nM MDA/microg protein) islets. In all, 500 islet equivalents (IE) of treated or control group islets were transplanted to BALB/c mice rendered diabetic with STZ. All animals showed a normal glucose clearance following a glucose load. Graft functionality was confirmed by normoglycemia (fasting plasma glucose: fpg < 150 mg/dl) after transplantation and reappearing hyperglycemia (fpg > 200 mg/dl) following removal of the graft. Suboptimal islet transplantation using 250 IE suggests that the grafted islet mass was inadequate for diabetes reversal. In addition, no significant differences were observed in the islet insulin content between the three groups following cryopreservation of the islets at -196 degrees C. Our studies indicate that taurine pretreatment and its continued presence during islet cryopreservation improves the postthawing viable recovery of islets. PMID- 11437071 TI - Reduction in primary nonfunction of syngeneic islet transplants with nordihydroguaiaretic acid, a lipoxygenase inhibitor. AB - To study the effectiveness of a lipoxygenase inhibitor, nordihydroguaiaretic acid (NDGA), in the reduction of primary nonfunction, an insufficient number of syngeneic islets were transplanted underneath the renal capsule with NDGA administered daily for 4 weeks. After transplantation of the 150 islets, the decrement of blood glucose levels was significantly faster in the mice that had received NDGA than in the mice that had received no drug at all or dimethyl sulfoxide (DMSO) (p < 0.005, p < 0.05). The mean duration of temporary posttransplant hyperglycemia was 22.3 +/- 3.2 (n = 10), 35.9 +/- 2.3 (n = 14), and 33.7 +/- 4.1 (n = 6) days for the respective groups. The diabetic mice that received 300 islets had their blood glucose levels decrease faster than those that received 150 islets (19.7 +/- 1.6 vs. 35.9 +/- 2.3 days, n = 14. p < 0.0001). There was no significant difference in the blood glucose reducing effect between the mice that received 150 islets with NDGA and the mice that received 300 islets [22.3 +/- 3.2 (n = 10) vs. 19.7 +/- 1.6 (n = 14) days, p > 0.05]. The insulin content of the graft from the mice treated with 150 islets and NDGA (3.02 +/- 0.24 microg, n = 4) was higher than that from the mice that received 150 islets but no treatment (1.10 +/- 0.26 microg, n = 15, p < 0.005) or that had been treated with DMSO (1.21 +/- 0.30 microg, n = 4, p <0.05). The insulin content of the pancreas remnant had no significant differences among the three groups. The net glucose-stimulated insulin secretion was 0.82 +/- 0.14 vs. 0.20 +/- 0.10 microIU/islet x 60 min (n = 8, p < 0.005) and 0.59 +/- 0.08 vs. 0.04 +/- 0.02 microIU/islet x 60 min (n = 8, p < 0.0001) for islets cultured without NDGA vs. with NDGA at 1 and 2 weeks, respectively. However, the insulin content of the cultured islets was similar between the two groups for up to 2 weeks of incubation (at 1 week: 0.71 +/- 0.01 vs. 0.67 +/- 0.04 ng/islet, n = 8, p > 0.05; at 2 weeks: 0.71 +/- 0.02 vs. 0.80 +/- 0.07 ng/islet, n = 8, p > 0.05). Serum leukotriene B4 (LTB4) concentrations before and between the fifth and seventh days after transplantation were determined. For diabetic mice that received 150 islets, serum LTB4 levels were 25,835 +/- 3,335 and 27,631 +/- 3,136 pg/ml (n = 4, p > 0.05). For diabetic mice that received 150 islets and NDGA, the corresponding figures were 22,401 +/- 2,706 pg/ml and 27,530 +/- 2,190 pg/ml (n = 8, p > 0.05). The graft histology revealed viable islet cells and networks of close vascular structures around the islets and did not reveal microscopic differences among the samples of all four groups. In conclusion, our data revealed that daily administration of NDGA for 4 weeks enhanced isoislet engraftment and preserved three times more mass of the islet beta cells in the isografts. This result indicates that NDGA reduces primary nonfunction of islet syngeneic grafts in diabetic mice. PMID- 11437072 TI - Poly-L-Lysine induces fibrosis on alginate microcapsules via the induction of cytokines. AB - Alginate-poly-L-lysine (PLL) microcapsules can be used for transplantation of insulin-producing cells for treatment of type I diabetes. In this work we wanted to study the inflammatory reactions against implanted microcapsules due to PLL. We have seen that by reducing the PLL layer, less overgrowth of the capsule is obtained. By incubating different cell types with PLL and afterwards measuring cell viability with MTT, we found massive cell death at concentrations of PLL higher than 10 microg/ml. Staining with annexin V and propidium iodide showed that PLL induced necrosis but not apoptosis. The proinflammatory cytokine, tumor necrosis factor (TNF), was detected in supernatants from monocytes stimulated with PLL. The TNF response was partly inhibited with antibodies against CD14, which is a well-known receptor for lipopolysaccharide (LPS). Bactericidal permeability increasing protein (BPI) and a lipid A analogue (B-975), which both inhibit LPS, did not inhibit PLL from stimulating monocytes to TNF production. This indicates that PLL and LPS bind to different sites on monocytes, but because they both are inhibited by a p38 MAP kinase inhibitor, they seem to have a common element in the signal transducing pathway. These results suggest that PLL may provoke inflammatory responses either directly or indirectly through its necrosis inducing abilities. By combining soluble PLL and alginate both the toxic and TNF inducing effects of PLL were reduced. The implications of these data are to use alginate microcapsules with low amounts of PLL for transplantation purposes. PMID- 11437073 TI - Influence of donor age on mouse islet characteristics and transplantation. AB - Old donor age has been considered as a risk factor and relative contraindication for transplantation. This study was designed to investigate the influence of donor age on islet characteristics and transplantation. Islets isolated from 8 (I A)-, 32 (I-B)-, or 64 (I-C)-week-old C57BL16 mice were studied for number, size, insulin content, and secretion. After syngeneically transplanting 300 islets under the kidney capsule of streptozotocin-diabetic mice (R-A. R-B, and R-C, respectively), we measured recipients' metabolic parameters as well as the beta cell mass and insulin content of the graft. Eight-week-old donors had better glucose tolerance than 32- and 64-week-old donors. However, 64-week-old donors had more pancreatic insulin content than 8- and 32-week-old donors. I-B and I-C were greater in number, larger in size, and higher in insulin content than I-A. But perifusion study showed I-C secreted less insulin, albeit with a similar stimulation index compared with that of I-A and I-B. After transplantation, the fall of blood glucose in R-C was faster than that in R-A and R-B. At 12 weeks, the recipients' blood glucose, body weight, HbA1c, and the beta-cell mass and insulin content of the graft were comparable in all groups. However, R-C had better glucose tolerance than R-A. During follow-up, R-A and R-B maintained lifelong normoglycemia and their glucose tolerance did not deteriorate. These data indicate that islets isolated from donors with different ages have different characteristics and effects on transplantation. The islets isolated from aged donors are functioning well and can be a potential source for transplantation; however, because we transplanted a large islet mass from the aged donors, the role of the islet dose needs to be further clarified. PMID- 11437074 TI - Role of pancreatic polypeptide as a market of transplanted insulin-producing fetal pig cells. AB - Transplantation of insulin-producing fetal pancreatic tissue into diabetic recipients has been shown to normalize blood glucose levels after several months. This time period is required for the growth and maturation of the fetal tissue so insulin levels cannot be used as a marker of graft function while the beta-cell is immature. Therefore, we have examined the use of another pancreatic endocrine hormone, pancreatic polypeptide (PP), to monitor graft function. The cell that produces this hormone has been shown to be the first mature endocrine cell in the fetal pancreas. Fetal pig pancreatic tissue, both in the form of 1 mm3 explants and islet-like cell clusters (ICCs), was transplanted into immunodeficient SCID mice and the levels of PP and insulin were measured in plasma and in the graft for up to 12 weeks. PP was detected in the untransplanted explants (0.58 pmol/mg) and ICCs (0.06 pmol/ICC) and the PP to insulin ratio was 2.7% and 5.8%, respectively. PP (but not porcine C-peptide, a marker of insulin secretion) was detectable in the plasma of SCID mice from 4 days to 3 weeks after transplantation, but not thereafter. The highest values were obtained at 4 days to 1 week. In the grafted tissue PP and insulin were present at all time points and the ratio of PP to insulin was 59%, 87%, 75%, 56%, 7%, 8%, and 7% at 4 days, 1, 2, 3, 6, 9, and 12 weeks, respectively. The decline in PP levels 3 weeks after transplantation was associated with beta-cell development in the graft. PP was also secreted by fetal pig pancreatic explants transplanted into diabetic NOD/SCID mice, with plasma levels measurable in the first week after the tissue was grafted. In immunocompetent BALB/c mice transplanted with the tissue, PP was detectable in plasma for 2 days after transplantation but not at 4 days, when cellular rejection commenced, or thereafter. We conclude that plasma PP levels can be used as a marker of the viability of fetal porcine pancreatic tissue in the first 3 weeks after it is transplanted into mice. These findings may have relevance to fetal pancreatic tissue transplanted into humans if suitable techniques can be developed to separate pig from human PP. PMID- 11437075 TI - Enhanced survival of porcine neural xenografts in mice lacking CD1d1, but no effect of NK1.1 depletion. AB - Transplantation of embryonic porcine neurons may restore neurological function in patients with Parkinson's disease, if immunological rejection could be prevented. This study was performed to investigate the role of natural killer cells (NK cells) and NK1.1+ T cells (NK T cells) in the rejection of neural xenografts. A cell suspension was prepared from the ventral mesencephalon of 26-27-day-old pig embryos, and 2 microl was implanted in the right striata of mutant CD1d1 null (CD1.1-/-) mice, NK1.1-depleted mice, and controls. The CD1.1-/- mice are deficient in NK T cells and the antigen-presenting molecule CD1d1. Graft survival and host responses were determined immunohistochemically using markers for dopamine neurons, CD4-, CD8- cells, microglia, and macrophages. At 2 weeks, the grafts were significantly larger in CD1.1-/- mice, 0.09 +/- 0.02 microl (mean +/- SEM), compared with controls, 0.05 +/- 0.01 microl. There was no significant difference between NK1.1-depleted mice, 0.02 +/- 0.01 microl, and controls. At 5 weeks, two grafts were still present in the CD1-/- mice, whereas only scars remained in the controls and in the NK1.1-depleted mice. Immune reactions were strong at 2 weeks and less pronounced at 5 weeks in all groups. Microglial activation was lower in NK-depleted mice than in the controls at 2 weeks. In contrast to organ xenografting, NK1.1+ cells do not seem to be important mediators of the rejection of discordant cellular neural xenografts. However, our results suggest that the antigen-presenting molecule CD1d1 may be involved in the rejection process. PMID- 11437076 TI - Remyelination of demyelinated CNS axons by transplanted human schwann cells: the deleterious effect of contaminating fibroblasts. AB - Areas of demyelination can be remyelinated by transplanting myelin-forming cells. Schwann cells are the naturally remyelinating cells of the peripheral nervous system and have a number of features that may make them attractive for cell implantation therapies in multiple sclerosis, in which spontaneous but limited Schwann cell remyelination has been well documented. Schwann cells can be expanded in vitro, potentially affording the opportunity of autologous transplantation; and they might also be spared the demyelinating process in multiple sclerosis. Although rat, cat, and monkey Schwann cells have been transplanted into rodent demyelinating lesions, the behavior of transplanted human Schwann cells has not been evaluated. In this study we examined the consequences of injecting human Schwann cells into areas of acute demyelination in the spinal cords of adult rats. We found that transplants containing significant fibroblast contamination resulted in deposition of large amounts of collagen and extensive axonal degeneration. However, Schwann cell preparations that had been purified by positive immunoselection using antibodies to human low affinity nerve growth factor receptor containing less than 10% fibroblasts were associated with remyelination. This result indicates that fibroblast contamination of human Schwann cells represents a greater problem than would have been appreciated from previous studies. PMID- 11437077 TI - Posterior segment approach for subretinal transplantation or injection in the canine model. AB - A posterior segment approach for cell transplantation or injection into the subretinal space of the dog has been developed. Controlled penetration to the subretinal space was achieved using a 29-gauge injection cannula, either blunted or with a 30 degrees sharpened bevel, and partially ensheathed with moveable plastic tubing. Depending on the injection volume used, the retina detached, and the fluid was reabsorbed within 1-3 weeks, although for smaller volumes the retina reattached within a matter of days. The optimal injection volume used was between 100 and 150 microl, or two injections of 55 microl each. By ophthalmoscopy following the surgery, it was possible to serially monitor the injection site and retinal bleb through fundus photography. Light microscopy demonstrates the distribution of stable, viable RPE cells in the subretinal space up to 6 months. The transplantation technique developed for the dog is atraumatic and free from any major surgical or clinical complications. It can be readily used to deliver cells or fluids to localized regions of the subretinal space. PMID- 11437078 TI - Probing enhanced cytochrome P450 2B1/2 activity in rat hepatocyte spheroids through confocal laser scanning microscopy. AB - Cytochrome P450 (CYP450) enzymes are essential for xenobiotic metabolism. Although CYP450s are found in many tissues, CYP2B1/2 are primarily expressed in the rat liver. The constitutive expression in vivo of CYP2B1/2 is low but it is induced in the presence of various drugs such as phenobarbital (PB). In this study, CYP2B1/2 activity in cultured hepatocytes was assessed in situ with the introduction of a fluorogenic substrate, pentoxyresorufin. The product of 7 pentoxyresorufin-O-dealkylation (PROD), which is catalyzed specifically by CYP2B1/2, was detected using confocal laser scanning microscopy (CLSM). Primary hepatocytes cultured as monolayers on collagen-coated surfaces exhibited background PROD activity and minimal PB inducibility after 4 days in culture. In contrast, rat hepatocytes organized in compacted aggregates, or spheroids, exhibited higher levels of PROD activity and retained their ability for PB induction. The results from the CLSM analysis were verified by RT-PCR and Western immunoblotting analysis. Furthermore, CLSM in conjunction with image processing techniques and three-dimensional reconstruction revealed the localization of enhanced PROD activity in the center of spheroids. The results support the use of CLSM as a powerful tool for investigating CYP2B1/2 activity in cultured rat hepatocytes. PMID- 11437079 TI - Liver-derived dendritic cells induce donor-specific hyporesponsiveness: use of sponge implant as a cell transplant model. AB - Spontaneously accepted mouse liver allografts are capable of protecting subsequently transplanted donor organs from rejection; however, the underlying mechanisms are unclear. Dendritic cells (DC) residing in liver grafts are likely important in tolerance induction. DC propagated from mouse liver with GM-CSF are phenotypically and functionally immature. They are poor allostimulators in MLR and prolong survival of pancreatic islet allografts. It has been problematic to perform mechanistic studies in an islet transplant model because of difficulties in obtaining sufficient graft infiltrating cells. In this study, we used a sponge allograft model [i.e.. a subcutaneously implanted sponge matrix loaded with B10 (H2b) spleen cells]. To investigate the influence of administration of donor (B10) liver-derived DC on alloimmune reactivity of C3H (H2k) hosts, sponge graft infiltrating cells (SGIC) and recipient spleen cells were isolated, and their immunophenotype and donor-specific cytotoxic T lymphocyte (CTL) activity were examined. The results illustrate that donor-specific CTL activity of T cells are lower in recipients that had received systemic treatment with liver-derived immature DC, associated with a decrease in CD8+ cell population and an increase in Gr-1+ cells in SGIC, compared with recipients treated with mature bone marrow (BM)-derived DC. Interestingly, administration of liver DC directly into the sponge did not inhibit T cell responses. These data suggest that systemic administration of donor liver DC induces donor-specific hyporesponsiveness, probably not by direct inhibition of graft infiltrating T cells. The increased Gr 1+ cells may play immune regulatory roles in induction of host donor-specific hyporesponsiveness. PMID- 11437080 TI - Validity and sensitivity to change of the extended Glasgow Outcome Scale in mild to moderate traumatic brain injury. AB - Using a structured outcome interview, this study addressed the validity and sensitivity to change of the Glasgow Outcome Scale (GOS) and the Extended GOS (GOSE) in a prospective study of patients who sustained mild (n = 30) to moderate (n = 13) traumatic brain injury (TBI) or general trauma (n = 44). The patients were recruited from the emergency center or inpatient units of Ben Taub General Hospital and invited to participate in follow-up examinations at 3 and 6 months. Using a series of functional outcome measures, assessment of affective status, and neuropsychological tests as criteria, the validity of the GOSE generally exceeded the GOS. Analysis of the outcome data for the patients who completed both the 3-month and 6-month assessments disclosed that the GOSE was more sensitive to change than the GOS. Comparison of the 3-month outcome data disclosed that the GOSE and GOS scores did not differ for the TBI and general trauma groups. These findings lend further support for utilization of the GOSE in clinical trials when it is based on a structured interview. PMID- 11437081 TI - Abnormal cerebral blood volume in regions of contused and normal appearing brain following traumatic brain injury using perfusion magnetic resonance imaging. AB - Following traumatic brain injury, there may be secondary alterations in cerebrovascular parameters leading to ischemia and further cellular damage. To assess possible subacute hemodynamic disturbances following traumatic brain injury, we used conventional and perfusion magnetic resonance imaging (MRI) in 18 patients, on average 10 days following injury. Six of the 18 patients had focal contusions or edema visible on conventional MRI. These six patients had a significantly reduced normalized regional cerebral blood volume (rCBV) in the regions of focal pathology compared to equivalent areas in control subjects (patients 0.47 +/- 0.20 [means +/- SD], controls 1.02 +/- 0.11, p < 0.001). In addition, four of these six patients had an increased rCBV (outside control range) in the region of normal appearing brain immediately surrounding the contusion. These six patients were more significantly injured and had a worse clinical outcome compared to the remaining patients (p = 0.004,p = 0.03, respectively). There were five patients who had a region of reduced rCBV (outside control range) in a quadrant of normal appearing white matter, away from any visible abnormality, who were not more significantly injured than the remaining patients but went on to have a significantly poorer clinical outcome (p = 0.27, p = 0.01, respectively). Traumatic brain injury is a heterogeneous insult causing a variety of pathology, not all of which is visible using conventional imaging methods. The current study has shown that regions of both normal appearing and contused brain may have an abnormal rCBV and that alterations in rCBV may play a role in determining the clinical outcome of patients. PMID- 11437082 TI - Bedside monitoring of cerebral blood flow by transcranial thermo-dye-dilution technique in patients suffering from severe traumatic brain injury or subarachnoid hemorrhage. AB - Bedside measurement of cerebral blood flow (CBF) represents an important feature in monitoring of neurointensive care patients which is hard to establish. Therefore, we adopted a recently described thermo-dye-dilution-based approach for monitoring CBF in patients suffering from severe cerebral insults, that is, traumatic brain injury (TBI) or subarachnoid hemorrhage (SAH). Combined fiberoptic-thermistor catheters were placed in one jugular venous bulb and in the abdominal aorta of 16 patients. Following central venous injection of a 50-mL bolus of precooled indocyanine green (ICG) solution, CBF was determined as a function of the mean transit times of coldness and dye. In addition, measurements of CBF using stable xenon-enhanced computerized tomography (sXe-CT) were simultaneously performed in 10 patients. A total of 272 thermo-dye-dilution measurements yielded 196 valid results, with CBF ranging from 26.2 to 144.8 mL 100 g(-1) min(-1). Reproducibility was fairly good, with the standard deviation within sets of repeated measurements being 6.3 mL 100 g(-1) min(-1) and 9.4 as the mean coefficient of variation. Simultaneously obtained values with sXe-CT displayed a good correlation (r = 0.843, p < 0.01); however, the thermo-dye dilution method consistently overestimated CBF. Data analysis using the Bland and Altman methodology revealed a large bias of 45.7 mL 100 g(-1) min(-1) with a +/- 2 SD range of 37 mL 100 g(-1) min(-1), indicating a rather poor agreement. The thermo-dye-dilution method proved a reasonably reproducible technique, enabling repeated long-term bedside measurements of CBF in neurointensive care patients with a minimum of time effort. However, a high failure rate was also noted, and consistent overestimation of perfusion was observed in comparison to sXe-CT measurements. Although the thermo-dye-dilution technique has been successfully validated in patients with normal neurovascular function, its applicability for bedside monitoring of CBF appears uncertain in patients suffering from severe TBI or SAH. PMID- 11437083 TI - The immunophilin ligand FK506 attenuates axonal injury in an impact-acceleration model of traumatic brain injury. AB - The immunophilin ligand, cyclosporin A (CsA), is effective in reducing the axonal damage associated with traumatic brain injury (TBI). Based upon extensive ultrastructural and immunohistochemical studies, the neuroprotection afforded by CsA appeared to be mediated via mitochondrial protection, specifically, the prevention of mitochondrial swelling and inhibition of mitochondrial permeability transition (MPT). However, the potential that CsA could also be neuroprotective via the immunophilin-mediated inhibition of the protein phosphatase, calcineurin (CN) has not been directly assessed. To address this issue, the current study assessed the ability of FK506, another immunophilin ligand that inhibits CN with no effect on MPT, to attenuate axonal damage in a rat impact-acceleration model of TBI. Traumatic axonal injury (TAI), detected via an antibody against beta amyloid precursor protein (APP), a specific marker of axonal injury, was significantly reduced at 24 hr postinjury in Sprague-Dawley rats receiving intravenous FK506 (2 mg/kg; n = 5) 30 min prior to injury compared to vehicle controls (n = 3). While not rejecting the established efficacy of CsA in providing neuroprotection via its targeting of MPT, this study does underscore the potential importance of CN in the progressive pathobiology of TAI, suggesting that CN may constitute another important therapeutic target. PMID- 11437084 TI - Age-Dependent vasopressinergic modulation of Noc/oFQ-induced impairment of NMDA cerebrovasodilation after brain injury. AB - This study was designed to characterize the role of vasopressin in nociceptin/orphanin FQ (NOC/oFQ)-induced impairment of NMDA cerebrovasodilation after fluid percussion brain injury (FPI) as a function of age in newborn (1-5 days old) and juvenile (3-4 weeks old) pigs equipped with a closed cranial window. Previous studies have observed that NOC/oFQ is released into CSF and contributes to impaired NMDA induced pial artery dilation following FPI to a greater extent in newborn versus juvenile pigs. Topical vasopressin (40 pg/mL), a concentration approximating that observed in CSF following FPI in the newborn, increased CSF NOC/oFQ from 69 +/- 3 to 102 +/- 8 pg/mol under non-FPI conditions. CSF NOC/oFQ was elevated within 60 min of FPI (70 +/- 3 to 444 +/- 51 pg/mL), but release was attenuated by MEAVP, a vasopressin antagonist, in the newborn (71 +/- 3 to 146 +/- 11 pg/mL). CSF vasopressin and NOC/oFQ were not elevated as greatly in the juvenile following FPI and MEAVP correspondingly did not attenuate CSF NOC/oFQ release as much as in the newborn. Under noninjury conditions, vasopressin (40 pg/mL) coadministered with NMDA (10(-8), 10(-6) M) attenuated pial dilation to this excitatory amino acid (9 +/- 1% and 16 +/- 1% vs. 3 +/- 1% and 5 +/- 2%). Following FPI in the newborn, NMDA-induced pial artery dilation was reversed to vasoconstriction, and both NOC/oFQ and vasopressin receptor antagonists partially prevented these alterations (9 +/- 1%) and 16 +/- 1%, sham control; -7 +/- 1% and -12 +/- 1%, FPI; -2 +/- 1% and -3 +/- 1%, FPI-NOC/oFQ antagonist; and 1 +/- 1% and 4 +/- 1%, FPI-vasopressin antagonist). NMDA-induced pial dilation was only attenuated following FPI in the juvenile and modestly restored by NOC/oFQ and vasopressin receptor antagonists. These data show that vasopressin, in concentrations present in CSF following FPI, contributes to the release of CSF NOC/oFQ following such an insult. The greater release of vasopressin following FPI in the newborn contributes to the corresponding greater release of NOC/oFQ in the newborn versus the juvenile. Moreover, vasopressin also contributes to the impairment of NMDA cerebrovasodilation after brain injury to a greater extent in newborn versus juveniles. These data suggest that vasopressin modulates NOC/oFQ-induced impairment of NMDA cerebrovasodilation after brain injury in an age-dependent manner. PMID- 11437085 TI - Extracellular superoxide dismutase overexpression improves behavioral outcome from closed head injury in the mouse. AB - Oxidative stress is known to play an important role in the response of brain to traumatic insults. We tested the hypothesis that increased extracellular superoxide dismutase (EC-SOD) expression can reduce injury in a mouse model of closed head injury. Neurologic, cognitive, and histologic outcomes were compared between transgenic mice exhibiting a fivefold increase in EC-SOD activity and wild-type littermate controls. Severe or moderate transcranial impact was induced in anesthetized and physiologically controlled animals. After severe impact, transgenic mice had better neurological outcome at 24 hr postinjury (p = 0.038). Brain water content was increased, but there was no difference between groups. Moderate impact resulted in predominantly mild neurologic deficits in both groups at both 24 hr and 14 days postinjury. Morris water maze performance, testing cognitive function at 14-17 days after trauma, was better in EC-SOD overexpressors (p = 0.018). No differences were observed between groups for histologic damage in hippocampal CA1 and CA3. We conclude that EC-SOD has a beneficial effect on behavioral outcome after both severe and moderate closed head injury in mice. Because EC-SOD is believed to be predominantly located in the extracellular space, these data implicate an adverse effect of extracellular superoxide anion on outcome from closed head injury. PMID- 11437086 TI - Influence of posttraumatic hypoxia on behavioral recovery and histopathological outcome following moderate spinal cord injury in rats. AB - Pulmonary dysfunction leading to secondary hypoxia is a common complication of spinal cord injury (SCI). The purpose of this study was to clarify the behavioral and histopathological consequences of posttraumatic hypoxia in an established model of traumatic SCI. Forty-five female Sprague-Dawley rats were randomly assigned to one of four groups, including (1) laminectomy and normoxia (n = 10), (2) laminectomy and hypoxia (n = 11), (3) NYU weight-drop and normoxia (n = 12), and (4) NYU weight-drop and hypoxia (n = 11). For these studies, a moderate injury was induced by adjusting the height of the weight drop (10 g) to 12.5 mm above the exposed spinal cord (T10). Immediately after injury, PaO2 in the hypoxic rats was kept between 30 and 35 mm Hg for 30 min. PaO2 in the normoxic group was maintained over 100 mm Hg, while PaCO2 in all rats was maintained at 35 40 mm Hg. The behavior of the rats was checked every 7 days using the Basso, Beattie, and Bresnahan (BBB) locomotor rating scale. Rats were sacrificed at 8 weeks for quantitative histopathological analysis of lesion areas. During the hypoxic insults, the mean arterial blood pressure dropped in both sham control and weight-drop rats (p < 0.01). At the end of the 8-week monitoring period, BBB scores were 12.5 +/- 3.1 (mean +/- SEM) and 14.2 +/- 3.4 in the normoxic and hypoxic traumatized rats, respectively. No significant difference between the traumatized groups was documented with BBB monitoring. In contrast, the percent of gray matter necrosis at the impact epicenter was significantly increased in hypoxic versus normoxic SCI rats (p < 0.01). These data demonstrate that posttraumatic hypoxia complicated by mild hypotension aggravates the histopathological consequences of SCI and further emphasize the need to control for secondary hypoxic insults after experimental and clinical SCI. Potential explanations for the lack of a correlation between the behavioral and histopathological findings are discussed. PMID- 11437087 TI - Presentation of a previously asymptomatic Chiari I malformation by a flexion injury to the neck. AB - Flexion injury and/or whiplash injury to the neck in car accidents are usually trivial injuries with no serious neurological deficits. Our intention was to point out the importance to proceed with diagnostic procedures if neurological deficits do occur in order to reveal the true cause of the deficit. The paper presents the case of a 35-year-old woman who sustained a flexion injury to the neck. A relatively trivial injury to the neck promoted a progressive neurological deterioration. The standard diagnostic procedures (x-ray, computed tomography scan) were normal. Further diagnostics with magnetic resonance imaging was required to reveal an underlying Chiari I malformation. Finally, the operative decompression of the craniocervical junction was performed. Following the surgical treatment, the patient's clinical symptoms regressed. One year after her discharge, she remains in good physical condition. To our knowledge, this case is the first report of the manifestation of Chiari I malformation in the adult as a result of a flexion or whiplash injury of the neck. This unusual case suggests that in a trivial flexion injury to the neck sustained in a car accident, which presents with serious neurological dysfunction, and where the standard diagnostic procedures are normal, the possibility of underlying congenital abnormality, such as Chiari I malformation should be considered. PMID- 11437088 TI - Projecting severe sequelae of injection-related hepatitis C virus epidemic in the UK. Part 1: Critical hepatitis C and injector data. AB - BACKGROUND: Hepatitis C is transmitted by transfusion of unscreened blood, through injecting drugs, from mother-to-child and, on occasion, sexually. Transmission generally requires that the infector is hepatitis C virus (HCV) RNA positive, a 'carrier'. About three-quarters of injectors who are hepatitis C antibody positive are HCV-RNA positive and so infectious to others. Incubation periods from HCV infection to cirrhosis and hepatocellular carcinoma are even longer than from HIV infection to AIDS, being counted in decades; they depend on age, gender, alcohol consumption and co-infection with other viruses. We identify 25 data sources that are available, or required, for projecting the severe sequelae of the injection-related hepatitis C epidemic. DATA SOURCES: Three data sources relate to hepatitis C diagnosis: register of confirmed HCV infections (with initial of first name + soundex of surname + date of birth + gender = master index, exposure category, year of starting to inject, and region); surveys of HCV test-uptake by injectors and others; documentation of pregnancy and its outcome in HCV-infected women (injectors and others). Four data sources relate to HCV prevalence and incidence among injectors and others: anonymous testing for HCV antibodies in blood or saliva (for sentinel groups ranging from new blood donors, pregnant women, patients awaiting kidney transplantation, non-injector prisoners, health-care workers, non-injector heterosexuals attending genitourinary medicine clinics; to injectors in the community, at drug treatment centres or in prison); historical data on HCV prevalence in injectors; HCV incidence studies in injectors; and uptake of harm reduction measures--frequency of sharing and methadone substitution--by injectors. Key reporting problems in HCV incidence studies, which inhibit checks on the convenient exponential assumption for time from start of injecting to hepatitis C infection, are discussed. Nine critical data sources are identified for monitoring the late sequelae of hepatitis C carriage, its investigation and treatment: linkage surveillance, for example by master index, to identify deaths, hospitalisations or cancer registrations among confirmed HCV infections; surveys of HCV status among patients who undergo liver biopsy, are newly diagnosed with cirrhosis or are newly diagnosed with liver cancer; surveys of liver-biopsy rate in HCV infected injectors and others; uptake and outcome of interferon + ribavirin in the treatment of hepatitis C carriers; cohort studies of HCV progression; sample surveys of genotype in HCV-infected injectors, and others; acute hepatitis B infections and uptake of hepatitis B immunisation by injectors; liver transplantation in HCV-infected patients; and hepatitis C-status and other risk factors in deaths from cirrhosis or liver cancer, to determine whether they are HCV and injector-related. Finally, nine critical data sources are identified for quantitative understanding of the underlying injector epidemic: drug misuse databases plus capture-recapture methods to assess number of injectors, drug related deaths by region to assess injector numbers; number of HIV-infected injectors; HIV progression in injectors; overdose and other causes of death in injectors; expert opinion on injector incidence historically, plus survey information on age-distribution at initiation and duration of injector careers; injector incidence historically inferred from hepatitis C infected blood donors; age-distribution of current injectors and at initiation, as a check on the assumptions made in stochastic simulation about injector incidence and 'outcidence' from injecting historically; mortality of former injectors; and general population or other survey ratios of surviving ever-injectors to injectors in the last 5 years, last year and currently, as a check on simulations. RECOMMENDATIONS: We recommend a common HCV diagnosis report form to improve ascertainment of risk-factor information, especially year of starting to inject--which is a key date epidemiologically. We also recommend updated surveys of current and former injectors' HCV-test uptake, or a denominator study that registers master index and risk factor information for all HCV testees. We recommend that injector surveys ask about typical frequency of needle sharing per 4 weeks in three distinct periods this year, last year and in the first year of injecting. We also recommend the location of stored historical samples from injectors to be tested retrospectively and anonymously for HCV antibodies. We recommend immediate attention to the uptake of, and response to, combination treatment by hepatitis C carriers who are former or recovering injectors. We rec PMID- 11437089 TI - Projecting severe sequelae of injection-related hepatitis C virus epidemic in the UK. Part 2: Preliminary UK estimates of prevalent injection-related hepatitis C carriers, and derivation of progression rates to liver cirrhosis by gender and age at hepatitis C virus infection. AB - BACKGROUND: In Part 2, we illustrate how available data can be used to obtain preliminary estimates for Scotland of prevalent injection-related hepatitis C carriers and of maternally hepatitis C virus (HCV)-infected infants. Novel approaches to reducing uncertainty about the number of Scotland's HCV infected children of injector parents are discussed in brief. Three approaches, one direct and two indirect, to estimating the number of current and ever-injectors are presented for England and Wales. METHODS: Diagnosed HCV infections in injectors and HCV test uptake by current injectors are combined with survey estimates for the ratio of ever-injectors to current injectors to estimate prevalent injection related hepatitis C carriers. Household surveys give direct but potentially biased estimates of the number of current and ever-injectors. Indirect estimates make use of hepatitis C diagnoses in injectors, HCV prevalence and test-uptake by injectors, or exploit international comparisons. We comment on key reporting problems that inhibit synthesis of HCV progression studies; and suggest how to derive preliminary gender-and-age specific progression rates to liver cirrhosis for use in projections. RESULTS: Preliminary estimates for Scotland of prevalent injection-related hepatitis C carriers are: central estimate 39,000, inner uncertainty 16,000-59,000; of maternally hepatitis C virus (HCV)-infected infants central estimate 260, uncertainty 110-1100; and for England and Wales estimates of the number of prevalent ever-injectors are central estimate 360,000, uncertainty 240,000-835,000. Both hepatitis C prevalence in injectors and estimated numbers of current injectors are similar in Australia, and England and Wales (but not so for Scotland), Australian work on projections of severe HCV sequelae from hepatitis C infections may therefore be a suitable starting point for projections for England and Wales. Australia anticipates a doubling in the number of persons living with hepatitis C cirrhosis from 8500 in 1997 to over 17,000 in 2010. DISCUSSION: Australian projections of severe HCV sequelae used progression rates that, for simplicity, were independent of gender and of age at HCV infection. Faster HCV progression for males, and their higher injector prevalence, means that the impact of HCV infection on, for example, liver cancer may be evident to a greater extent and earlier in males. PMID- 11437093 TI - Projecting cancer incidence and mortality using Bayesian age-period-cohort models. AB - BACKGROUND: We present a practical application of an age-period-cohort model in a Bayesian frame-work for making cancer-burden projections. METHODS: Second degree autoregressive smoothing was used on the age, period and cohort effects for estimating future incidence and mortality. RESULTS: We are able to demonstrate the feasibility, flexibility and strengths of this approach. Compared with previously used methods, it performed better for providing point estimates when past trends continued into the future. However, the extremely wide credible intervals need careful interpretation. DISCUSSION: Part of the uncertainty is attributable to the possible inadequacy of the model and not necessarily relevant in the prediction of what would happen if the present trends continue into the future. PMID- 11437094 TI - Alcohol intake and high density lipoprotein-cholesterol: comparison of food frequency questionnaire and diet record in a general population sample. AB - BACKGROUND: Moderate alcohol consumption is known to affect several cardiovascular risk factors, including high density lipoprotein-cholesterol (HDL C). Accurate measurement of alcohol intake in epidemiological studies is crucial for valid estimation of alcohol-related effects. METHODS: We compared two widely used alcohol-intake assessment methods, a food frequency questionnaire (FFQ) and a 7-day diet record (DR) regarding the association between alcohol intake and HDL C in the general population. RESULTS: In a representative sample (842 men, 1103 women) of the population of former West Germany (VERA study), 75.6% of the participants reported drinking alcoholic beverages in the FFQ, whereas the percentage was 84.9% according to the DR. The median daily alcohol intake of consumers was 10.3 and 12.1 g, as assessed by the FFQ and the DR, respectively. There was moderate agreement between reported consumption levels ranked by quintiles according to FFQ and DR (kappa = 0.49). With both approaches alcohol intake showed a strong positive association with HDL-C in linear regression models. The estimated increases in HDL-C (mg dL(-1)) per gram alcohol per day from the FFQ and DR were 0.15 [95% confidence interval (CI): 0.12-0.18] and 0.14 (95% CI: 0.11-0.17), respectively. When participants with different classification of the amount of alcohol consumed by FFQ and DR were excluded from the analysis, the association between alcohol intake and HDL-C was more pronounced. DISCUSSION: Our analyses suggest that both FFQ and DR lead to similar results regarding the alcohol-HDL-C association in the general population, despite major differences in the recorded prevalence of drinking and amount of alcohol consumed. Both methods may tend to under-estimate the true alcohol-HDL-C association. PMID- 11437095 TI - Subjective health status assessment: evaluation of the Italian version of the SF 12 Health Survey. Results from the MiOS Project. AB - BACKGROUND: SF-12 is a generic short form health survey, developed in the USA from the original SF-36. It produces two summary measures evaluating physical and mental self-perceived health that are interchangeable with those from the SF-36. SF-12 has been successfully tested in nine Western European countries on large samples of the general population, where it has proved its brevity, comprehensiveness, reliability, validity and cross-cultural applicability. The present analysis directly assesses the SF-12 for the first time in various Italian settings, including the general population and specific patient groups. METHODS: Data for this report were collected from five different samples; in four of them the SF-12 was used as a 'stand-alone' instrument, while in the other one (used as the reference) it was embedded in the SF-36. Descriptive statistics, Spearman's correlation coefficients, confirmatory factor analysis, ordinal uni- and multi-variate least squares regression model and covariance analysis were used to evaluate the summary measures in each sample, and across relevant subgroups. Studies were ordered according to the expected deviance, from the 'normal' health status of the reference group to the sample with the expected highest level of illness. RESULTS: Overall, more than 11,000 subjects were evaluated. Response rates ranged from 63 to 100%, while missing items accounted only for 0.2-8.2% of all items. Uni- and multi-variate analyses showed a positive association between both physical component summary (PCS) and mental component summary (MCS) scores and their respective items in all examined samples. MCS scores were fairly similar across all samples, with the only exception being patients recently discharged from hospital, whose subjective mental health perception was higher than expected and the highest of all (52.2). Finally, we found a substantial impact of ageing on physical health perception, while the MCS was shown to be less sensitive to the age effect. CONCLUSIONS: This analysis shows that the SF-12 has good validity, while some issues related to its most appropriate mode of administration and target groups might require further attention. PMID- 11437096 TI - Metastatic bone pain palliation with 89-Sr and 186-Re-HEDP in breast cancer patients. AB - AIM: The study evaluates the therapeutic efficacy of Strontium-89-chloride (89Sr) and 186Re-1,1-hydroxyethylidene diphosphonate (186Re-HEDP) in the palliation of painful bone metastases from breast cancer. PATIENTS AND METHODS: Fifty patients with painful multifocal bone metastases from breast cancer entered the study and were randomized into two groups according to the radiopharmaceutical used: 148 MBq 89Sr i.v. (Group A: 25 patients) and 1406 MBq 186Re-HEDP i.v. (Group B: 25 patients). Pain palliation was evaluated on the basis of the Wisconsin pain test improvement at two months and response was graded as complete, partial, minimal or absent. Hematological toxicity and side effects were reported according to WHO guidelines. RESULTS: The global response rate was 84% (21/25) for 89Sr and 92% (23/25) for 186Re-HEDP, respectively. The onset of pain palliation appeared significantly earlier in Group B (p < 0.0001). The duration of pain relief ranged from two months to 14 months (mean of 125 days with a median value of 120 days) in Group A and from one month to 12 months (mean of 107 days with a median value of 60 days) in Group B (p = 0.39). A moderate hematological toxicity was apparent in both groups. Platelet and white blood cell counts returned to baseline levels within 12 weeks after 89Sr administration and 6 weeks after 186Re-HEDP administration (p < 0.01). CONCLUSIONS: Both 89Sr and 186Re-HEDP are effective and safe in bone pain palliation in breast cancer with the latter showing a significantly faster onset of pain relief. PMID- 11437097 TI - Oncostatin M-induced growth inhibition and morphological changes of MDA-MB231 breast cancer cells are abolished by blocking the MEK/ERK signaling pathway. AB - Cytokine oncostatin M (OM) has profound effects on proliferation and differentiation of breast cancer cells. OM treated cells show reduced growth rate and differentiated phenotypes. The mechanisms underlying the OM growth-inhibitory activity in breast cancer cells have not been fully elucidated. In this study, we investigated the OM-elicited signaling pathways in breast cancer cell lines MDA MB231 and MCF-7. We show that OM rapidly activates the extracellular signal regulated kinase (ERK) and the signal transducer and activator of transcription (STAT) 1 and 3 in both cell lines. Intriguingly, OM-induced growth inhibition and morphological changes in MDA-MB231 cells are completely abolished by inhibitors to ERK upstream kinase MEK (nitrogen/extracellular-regulated protein kinase kinase), but the MEK inhibitors have little effects on OM growth-inhibitory activity in MCF-7 cells. In addition, expressions of the cyclin kinase inhibitors p21 and p27 are strongly induced by OM in MCF-7 cells, but their expression is only slightly increased by OM in MDA-MB231 cells. These data together demonstrate that the growth-inhibitory activity of OM can be mediated by different signaling pathways in a cell line-specific manner. While the MEK/ERK pathway is the predominant signaling pathway that leads to the growth inhibition of MDA-MB231 cells, activation of additional signaling pathways are necessary for OM to exert its growth-inhibitory activity in MCF-7 cells. PMID- 11437098 TI - Are medical oncologists biased in their treatment of the large woman with breast cancer? AB - PURPOSE: Obesity and breast cancer are common conditions that often coexist. Concerns of relative overdosing of chemotherapy in the large cancer patient have led clinicians to apply empiric dose reductions, 'cap' the body surface area (BSA) at 2 m2, or use ideal rather than actual body weight to calculate BSA. There are no data supporting or refuting these practices and their prevalence is unknown. We sought to determine the distribution of body size and prevalence of obesity in the breast cancer population of our cancer centre, and to determine clinician chemotherapy dosing practices in the era of modern adjuvant chemotherapy. PATIENTS AND METHODS: Women with invasive breast cancer receiving systemic therapy at our institution between 1980 and 1998 were identified and their recorded height and weight were used to calculate BSA and body mass index (BMI). We reviewed the first cycle adjuvant chemotherapy dosing practices from 1990-1998. The ideal dose of chemotherapy was calculated based on calculated BSA, and then contrasted with the actual dose received at cycle one. Discrepancies were recorded and categorized, using the largest single drug reduction if more than one drug was reduced. RESULTS: Mean BMI in the systemic therapy population was 26.4 +/- 5.3 kg/m2, 54% were overweight, 2% severely obese and 18% moderately so. Their mean BSA was 1.7 +/- 0.2 m2 and only 5% had a BSA > or = 2 m2. In the adjuvant chemotherapy subgroup, most patients received > or = 85% of their ideal dose. The mean dose reduction was 5.3 +/- 11.3% versus 9.9 +/- 11.3% in the BSA < 2 and > or = 2 m2 groups, respectively (p = 0.02), and 4.3 +/- 8.2% versus 6.7 +/ 13.1% in the BMI < 25 and > or = 25 kg/m2 groups, respectively (p = 0.008). While only 24% of chemotherapy dose reductions of > or = 15% were in the BSA > or = 2 m2 group, 76% were in the BMI > or = 25 kg/m2 group. CONCLUSIONS: Obesity is prevalent in this breast cancer population. BSA is not a sensitive index of large body size. We consistently detected more frequent empiric dose reductions at cycle one of adjuvant chemotherapy, with reductions of greater magnitude in the largest women (BSA > or = 2 m2) and those who were overweight (BMI > or = 25 kg/m2). PMID- 11437099 TI - Biophenotypes and survival of BRCA1 and TP53 deleted breast cancer in young women. AB - The aim of this study was to examine the loss of heterozygosity (LOH) of BRCA1 (17q21) and TP53 (17p13.1) in early-onset breast cancer patients; to correlate biopathological characteristics with molecular alterations; and to investigate the survival of LOH-related cancers. BRCA1 and TP53 LOH were evaluated in 78 early-onset breast cancers (< or = 40 years, Group 1) and 80 patients with age > 55 years (Group 2). Cases were characterized for multiple biological markers (ER, PR, proliferation index (PI), NEU and p53). LOH was carried out on microdissected paraffin embedded tissues; microsatellites D17S855 (BRCA1) and D17S786 (TP53) were amplified by fluorescent PCR and analyzed by an automated DNA sequencer. Early-onset breast cancers showed a higher frequency of ductal histotype (89.7% vs. 56.3% p < 0.001), node-positive (53.8% vs. 38.7%), larger size (p = 0.017), higher mitotic rate (p = 0.025), higher nuclear and final grade (p = 0.01 and p = 0.001, respectively). D17S855 LOH was 32.8% in group 1 vs. 21% in group 2; D17S786 LOH was 50.7% vs. 31.3% (p = 0.03), respectively. BRCA1 LOH was correlated with higher PI (p = 0.032) and higher p53 expression (p < 0.001) in group 1 and with higher NEU expression (p = 0.028) in group 2. TP53 LOH was correlated with p53 overexpression (p = 0.03) in group 1. A worse clinical outcome in early-onset LOH related cancers emerged from follow-up data: TP53 and BRCA1 LOH were associated with a shorter relapse free interval (RFI) (p = 0.03) and a poorer overall survival (OS) (p = 0.04), respectively. This study underlines different biological profiles in the two age groups investigated, probably reflecting different mechanisms of carcinogenesis. In accordance with adverse histopathological features in early-onset patients, LOH-related cancers have an unfavorable prognosis. PMID- 11437100 TI - Generation of reactive oxygen species by xanthine derivatives in MDA-MB-231 human breast cancer cells. AB - Theophylline reduces cell number in MDA-MB-231 cells through mechanisms over and above phosphodiesterase inhibition. In the current study, we used an intracellular fluorescent dye to show that theophylline and, to a much greater extent, 3-isobutyl-1-methylxanthine, evoke the generation of reactive oxygen species and also sensitize the cells to insult by other oxidants. Xanthine derivatives may therefore offer novel strategies for antitumor therapeutics. PMID- 11437101 TI - Methyl-substituted diindolylmethanes as inhibitors of estrogen-induced growth of T47D cells and mammary tumors in rats. AB - Diindolylmethane (DIM) is formed by acid catalyzed dimerization of the phytochemical indole-3-carbinol, and both compounds inhibit formation and/or growth of mammary tumors in rodents. In this study, we have investigated the aryl hydrocarbon receptor (AhR) agonist activity and inhibitory AhR-estrogen receptor crosstalk induced by the following methyl-substituted DIMs: 1,1'-dimethyl-, 2,2' dimethyl-, 5,5'-dimethyl-, 6,6'-dimethyl-, and 7,7'-dimethylDIM and 1,1',2,2' tetramethylDIM. The six compounds bound to the rat cytosolic AhR in a transformation assay but, at concentrations < or = 10 microM, exhibited minimal to non-detectable AhR agonist or antagonist activities associated with CYP1A1 induction. In contrast, the methyl-substituted DIMs inhibited estrogen-induced T47D human breast cancer cell growth and the four most active compounds (1,1'-, 2,2'-, 5,5'-dimethylDIM and 1,1',2,2'-tetramethylDIM) inhibited one or more estrogen-induced responses in the 21-day-old female B6C3F1 mice at a dose of 100 mg/kg/day (X3). Induction of hepatic CYP1A1-dependent activity was not observed at this high dose. The antitumorigenic activity of these compounds was examined in 7,12-dimethylbenz[a]anthracene-induced rat mammary tumor model in which the DIM analogs were orally administered (by gavage in corn oil) at a dose of 1 mg/kg/day (X10). 1,1'-DimethylDIM, 5,5'-dimethylDIM and 1,1',2,2'-tetramethylDIM significantly inhibited mammary tumor growth, and this was not accompanied by changes in organ/body weights or histopathology. These studies demonstrate that methyl-substituted DIMs are selective AhR modulators (SAhRMs) with potential for clinical treatment of breast cancer. PMID- 11437102 TI - Clinical significance of vascular endothelial growth factor-C (VEGF-C) in breast cancer. AB - Vascular endothelial growth factor-C (VEGF-C) is a specific ligand which induces lymphangiogenesis. We examined the expression of VEGF-C protein to determine its role in the progression of breast cancer. Immunohistochemical analysis revealed that VEGF-C was overexpressed in 39 of 98 breast cancer specimens (39.8%) but not in adjacent normal mammary glands. The expression of VEGF-C showed a significant correlation with lymphatic vessel invasion (p = 0.0004). It is noteworthy that the 5-year disease free survival rate of the VEGF-C positive group was significantly poorer than that of negative group (p = 0.0356). We suggest that as expression of VEGF-C is not implicated in lymphatic spread, it may prove to be a promising marker to predict the recurrence of breast cancer. PMID- 11437103 TI - Bifidobacterium longum as a delivery system for gene therapy of chemically induced rat mammary tumors. AB - A fundamental obstacle in cancer gene therapy is the specific targeting of therapy directly to a solid tumor, and no systemic delivery system yet exists. A strain of domestic bacteria, Bifidobacterium longum, which is nonpathogenic and anaerobic, selectively localized to and proliferated in 7,12 dimethylbenz[a]anthracene-induced rat mammary tumors after systemic application. We further ascertained the tumor specificity of genetically engineered, as well as wild-type, Bifidobacterium longum. This is the first demonstration that Bifidobacterium longum can be utilized as a specific gene delivery vector for gene therapy on solid breast tumors. PMID- 11437104 TI - Second primary breast cancer in men. PMID- 11437105 TI - Fas ligand expression in BRCA1-associated hereditary breast carcinoma clearly differs from that in sporadic breast carcinoma. AB - BRCA1-associated hereditary breast carcinomas (HBCs) are diagnosed at a younger age and are known to show biological aggressiveness such as a high histological grade, frequent aneuploidy, compared to sporadic breast carcinomas. However, results of studies on their prognosis have been controversial. We hypothesized that some factors such as a high incidence of cell death could offset the aggressiveness of BRCA1-associated HBCs, and therefore investigated Fas and Fas ligand (Fas L) expression in 19 BRCA1-associated HBCs and 56 age-adjusted control cases. Glandepithelial and myoepithelial cells in the mammary glands expressed Fas in high incidence, but all were negative for Fas L. Fas was expressed in 89.5% of BRCA1-associated HBCs and 94.4% of the controls and no significant differences could be established between the two groups. However, in 73.7% of BRCA1-associated HBCs, Fas L was clearly expressed in the infiltrating mononuclear cells, whereas this was observed in only 14.3% of the control cases and statistical significance was established between the two groups (p < 0.0001). These results strongly suggest that carcinoma cells in BRCA1-associated HBCs are more likely to be attacked by mononuclear cells via Fas L, and this may explain, at least in part, the discrepancy with respect to the prognosis. On the other hand, carcinoma cells also expressed Fas L significantly more often (p < 0.0001) in BRCA1-associated HBCs (52.6%) than in sporadic cases (3.6%). This can be considered a kind of counterattack against the mononuclear cells or, alternatively, may enhance the Fas-Fas L pathway in an autocrine/paracrine fashion. The clinical significance of Fas L expressing carcinoma cells remains to be elucidated. PMID- 11437106 TI - LV volume quantification via spatiotemporal analysis of real-time 3-D echocardiography. AB - This paper presents a method of four-dimensional (4-D) (3-D + Time) space frequency analysis for directional denoising and enhancement of real-time three dimensional (RT3D) ultrasound and quantitative measures in diagnostic cardiac ultrasound. Expansion of echocardiographic volumes is performed with complex exponential wavelet-like basis functions called brushlets. These functions offer good localization in time and frequency and decompose a signal into distinct patterns of oriented harmonics, which are invariant to intensity and contrast range. Deformable-model segmentation is carried out on denoised data after thresholding of transform coefficients. This process attenuates speckle noise while preserving cardiac structure location. The superiority of 4-D over 3-D analysis for decorrelating additive white noise and multiplicative speckle noise on a 4-D phantom volume expanding in time is demonstrated. Quantitative validation, computed for contours and volumes, is performed on in vitro balloon phantoms. Clinical applications of this spaciotemporal analysis tool are reported for six patient cases providing measures of left ventricular volumes and ejection fraction. PMID- 11437107 TI - Diffuse optical tomography of highly heterogeneous media. AB - We investigate the performance of diffuse optical tomography to image highly heterogeneous media, such as breast tissue, as a function of background heterogeneity. To model the background heterogeneity, we have employed the functional information derived from Gadolinium-enhanced magnetic resonance images of the breast. We demonstrate that overall image quality and quantification accuracy worsens as the background heterogeneity increases. Furthermore we confirm the appearance of characteristic artifacts at the boundaries that scale with background heterogeneity. These artifacts are very similar to the ones seen in clinical examinations and can be misinterpreted as actual objects if not accounted for. To eliminate the artifacts and improve the overall image reconstruction, we apply a data-correction algorithm that yields superior reconstruction results and is virtually independent of the degree of the background heterogeneity. PMID- 11437108 TI - Three-dimensional reconstruction of microcalcification clusters from two mammographic views. AB - Classification of benign/malignant microcalcification clusters is a major diagnostic challenge for radiologists. Clinical studies have revealed that the shape of the cluster, and the spatial distribution of individual microcalcifications within it, are important indicators of its malignancy. However, mammographic images of clustered microcalcifications confound their three-dimensional (3-D) distribution with image projection and breast compression. This paper presents a novel model-based method for reconstructing microcalcification clusters in 3-D from two mammographic views (cranio-caudal and medio-lateral oblique--"shoulder to the opposite hip" or lateral-medio). We develop a 3-D breast representation and a parameterised breast compression model which constraints geometrically the possible 3-D positions of a calcification in a two-dimensional image. Corresponding calcifications in the two views are matched using an estimate of the calcification volume. Both the geometric constraint and the matching criterion are utilized in the final reconstruction step to build the 3-D reconstructed clusters. Validation experiments are described using 30 clusters to verify the individual steps of the model, and results consistent with known ground truth are obtained. Some of the approximations in the model and future work are discussed in the concluding section. PMID- 11437109 TI - Automatic lung segmentation for accurate quantitation of volumetric X-ray CT images. AB - Segmentation of pulmonary X-ray computed tomography (CT) images is a precursor to most pulmonary image analysis applications. This paper presents a fully automatic method for identifying the lungs in three-dimensional (3-D) pulmonary X-ray CT images. The method has three main steps. First, the lung region is extracted from the CT images by gray-level thresholding. Then, the left and right lungs are separated by identifying the anterior and posterior junctions by dynamic programming. Finally, a sequence of morphological operations is used to smooth the irregular boundary along the mediastinum in order to obtain results consistent with those obtained by manual analysis, in which only the most central pulmonary arteries are excluded from the lung region. The method has been tested by processing 3-D CT data sets from eight normal subjects, each imaged three times at biweekly intervals with lungs at 90% vital capacity. We present results by comparing our automatic method to manually traced borders from two image analysts. Averaged over all volumes, the root mean square difference between the computer and human analysis is 0.8 pixels (0.54 mm). The mean intrasubject change in tissue content over the three scans was 2.75% +/- 2.29% (mean +/- standard deviation). PMID- 11437110 TI - Fast LV motion estimation using subspace approximation techniques. AB - Cardiac motion estimation is very important in understanding cardiac dynamics and in noninvasive diagnosis of heart disease. Magnetic resonance (MR) imaging tagging is a technique for measuring heart deformations. In cardiac tagged MR images, a set of dark lines are noninvasively encoded within myocardial tissue providing the means for measurement of deformations of the heart. The points along tag lines measured in different frames and in different directions carry important information for determining the three-dimensional nonrigid movement of left ventricle. However, these measurements are sparse and, therefore, multidimensional interpolation techniques are needed to reconstruct a dense displacement field. In this paper, a novel subspace approximation technique is used to accomplish this task. We formulate the displacement estimation as a variational problem and then project the solution into spline subspaces. Efficient numerical methods are derived by taking advantages of B-spline properties. The proposed technique significantly improves our previous results reported in [3] with respect to computational time. The method is applied to a temporal sequence of two-dimensional images and is validated with simulated and in vivo heart data. PMID- 11437111 TI - In vivo measurement of 3-D skeletal kinematics from sequences of biplane radiographs: application to knee kinematics. AB - Current noninvasive or minimally invasive methods for evaluating in vivo knee kinematics are inadequate for accurate determination of dynamic joint function due to limited accuracy and/or insufficient sampling rates. A three-dimensional (3-D) model-based method is presented to estimate skeletal motion of the knee from high-speed sequences of biplane radiographs. The method implicitly assumes that geometrical features cannot be detected reliably and an exact segmentation of bone edges is not always feasible. An existing biplane radiograph system was simulated as two separate single-plane radiograph systems. Position and orientation of the underlying bone was determined for each single-plane view by generating projections through a 3-D volumetric model (from computed tomography), and producing an image (digitally reconstructed radiograph) similar (based on texture information and rough edges of bone) to the two-dimensional radiographs. The absolute 3-D pose was determined using known imaging geometry of the biplane radiograph system and a 3-D line intersection method. Results were compared to data of known accuracy, obtained from a previously established bone-implanted marker method. Difference of controlled in vitro tests was on the order of 0.5 mm for translation and 1.4 degrees for rotation. A biplane radiograph sequence of a canine hindlimb during treadmill walking was used for in vivo testing, with differences on the order of 0.8 mm for translation and 2.5 degrees for rotation. PMID- 11437112 TI - Landmark-based elastic registration using approximating thin-plate splines. AB - We consider elastic image registration based on a set of corresponding anatomical point landmarks and approximating thin-plate splines. This approach is an extension of the original interpolating thin-plate spline approach and allows to take into account landmark localization errors. The extension is important for clinical applications since landmark extraction is always prone to error. Our approach is based on a minimizing functional and can cope with isotropic as well as anisotropic landmark errors. In particular, in the latter case it is possible to include different types of landmarks, e.g., unique point landmarks as well as arbitrary edge points. Also, the scheme is general with respect to the image dimension and the order of smoothness of the underlying functional. Optimal affine transformations as well as interpolating thin-plate splines are special cases of this scheme. To localize landmarks we use a semi-automatic approach which is based on three-dimensional (3-D) differential operators. Experimental results are presented for two-dimensional as well as 3-D tomographic images of the human brain. PMID- 11437113 TI - Simultaneous correction of ghost and geometric distortion artifacts in EPI using a multiecho reference scan. AB - A computationally efficient technique is described for the simultaneous removal of ghosting and geometrical distortion artifacts in echo-planar imaging (EPI) utilizing a multiecho, gradient-echo reference scan. Nyquist ghosts occur in EPI reconstructions because odd and even lines of k-space are acquired with opposite polarity, and experimental imperfections such as gradient eddy currents, imperfect pulse sequence timing, B0 field inhomogeneity, susceptibility, and chemical shift result in the even and odd lines of k-space being offset by different amounts relative to the true center of the acquisition window. Geometrical distortion occurs due to the limited bandwidth of the EPI images in the phase-encode direction. This distortion can be problematic when attempting to overlay an activation map from a functional magnetic resonance imaging experiment generated from EPI data on a high-resolution anatomical image. The method described here corrects for geometrical distortion related to B0 inhomogeneity, gradient eddy currents, radio-frequency pulse frequency offset, and chemical shift effect. The algorithm for removing ghost artifacts utilizes phase information in two dimensions and is, thus, more robust than conventional one dimensional methods. An additional reference scan is required which takes approximately 2 min for a matrix size of 64 X 64 and a repetition time of 2 s. Results from a water phantom and a human brain at 3 T demonstrate the effectiveness of the method for removing ghosts and geometric distortion artifacts. PMID- 11437114 TI - Comments on the filtered backprojection algorithm, range conditions, and the pseudoinverse solution. AB - The filtered backprojection (FBP) algorithm is widely used in computed tomography for inverting the two-dimensional Radon transform. In this paper, we analyze the processing of an inconsistent data function by the FBP algorithm (in its continuous form). Specifically, we demonstrate that an image reconstructed using the FBP algorithm can be represented as the sum of a pseudoinverse solution and a residual image generated from an inconsistent component of the measured data. This reveals that, when the original data function is in the range of the Radon transform, the image reconstructed using the FBP algorithm corresponds to the pseudoinverse solution. When the data function is inconsistent, we demonstrate that the FBP algorithm makes use of a nonorthogonal projection of the data function to the range of the Radon transform. PMID- 11437115 TI - Pregnancy, stress and wife assault: ethnic differences in prevalence, severity, and onset in a national sample. AB - Research using primarily hospital-based samples has suggested that pregnancy may put women at increased risk for wife assault, however, this research is largely limited by the lack of a comparison group of women who are not pregnant, and the failure to consider racial or ethnic differences in risk for violent victimization. The present study uses data from the 1992 National Alcohol and Family Violence Survey, a national probability sample of 1,970 individuals, to examine the prevalence, severity, and onset of wife assaults associated with pregnancy among Anglo and Hispanic families. The results indicated that pregnancy was associated with minor assaults among Hispanic women and severe assaults among Anglo women. Multivariate analyses revealed that among both Anglo and Hispanic families, there was no direct effect of pregnancy on risk for violent victimization after controlling for socioeconomic status, stressful life events, and age. PMID- 11437116 TI - Interpersonal and systemic aspects of emotional abuse at work: the target's perspective. AB - The most frequent form of workplace aggression is not physical, it is emotional and psychological in nature. Known by many names, emotional abuse at work is rapidly becoming recognized as pervasive and costly both in individual and organizational terms. Most of the research to date on emotional abuse at work has utilized survey and other quantitative methodologies in an effort to document the presence, prevalence, and impact of these behaviors. However, these methodologies are based on researchers' definitions and theories of what constitutes emotional abuse rather than on the meaning given to these experiences by the targets of these behaviors. A thorough understanding of this phenomenon requires a scholarly appreciation of the target's experience. Taking "feeling abused" as the criterion variable, this study examined target's experiences based on interviews with people who self-identifed as having experienced difficulties with a boss, coworker, or subordinate. The interpersonal aspects of emotional abuse focused on the nature of behaviors exhibited and the respondents' labeling of their experience. Consistent with elements of researchers' definitions, behaviors were defined as abusive when they were repetitive, resulted in injury or harm to target, and were experienced as a lack of recognition of the individual's integrity. Judgments of violation of standards of conduct and unsolicited nature of the behaviors were also related to respondents' experiences. Relative power differential was also an important element. However, contrary to researchers' definitions, actor intent was not central in defining the experience as abusive. The systemic aspect of emotional abuse was illustrated in the nature of organizational responding to concerns raised by respondents. These responses were of critical importance in respondents' labeling of their experiences as abusive. The focus on the meaning of the behaviors for the respondents provides an enriched picture of key definitional elements. Implications of the findings for future research are discussed. PMID- 11437117 TI - Abused women or abused men? An examination of the context and outcomes of dating violence. AB - The present study examines the controversial issue of whether women and men are equally abused in dating relationships. Undergraduate and graduate students (n = 874) completed a survey about their experiences and perpetration of psychological, sexual, and physical aggression within dating relationships. To enable a more contextualized understanding of these phenomena, motives for and outcomes of dating violence were also assessed. Women and men reported comparable amounts of overall aggression from dating partners, but differed in the types of violence experienced. Women were more likely to experience sexual victimization, whereas men were more often the victims of psychological aggression; rates of physical violence were similar across genders. Contrary to hypotheses, women were not more likely to use physical violence in self-defense than men. However, although both genders experienced similar amounts of aggressive acts from dating partners, the impact of such violence is more severe for women than men. PMID- 11437118 TI - Social reactions to rape victims: healing and hurtful effects on psychological and physical health outcomes. AB - In this study, 102 rape survivors were interviewed about the social reactions they received from family and friends post-rape. Results supported Ullman's (1996b) conclusion that the overall contribution of positive social reaction (e.g., providing support, listening, believing) on victims' recovery is negligible, but that negative social reactions (e.g., blaming) hinder recovery. In contrast to Ullman's (1996b) work, this research also examined whether rape victims have similar perceptions as to what constitutes a "positive" and "negative" social reaction. Results indicated that victims often agree as to what reactions are healing (positive), but that they do not agree as to what is hurtful (negative). By taking victims' perceptions into account, this study was able to compare the relative contributions of social reactions that were considered healing, social reactions that were considered hurtful, and the absence of social reactions. Results indicated that survivors who had someone believe their account of what happened or were allowed to talk about the assault- and considered these reactions to be healing-had fewer emotional and physical health problems than victims who considered these reactions hurtful, or victims who did not experience these reactions at all. Implications for future research on social reactions are discussed. PMID- 11437119 TI - A meta-analysis of rape education programs. AB - Meta-analysis of evaluation studies of rape education programs aimed at college students examined which program characteristics were related to participants' rape-supportive attitude change. Linear regression analyses revealed that: (a) published studies yielded greater attitude change than dissertations, presentations, or unpublished studies; (b) attitude change declined over time; and (c) men in mixed-gender groups experienced less attitude change after interventions than men in single-gender groups. Implications for the development of effective rape education programs are discussed. PMID- 11437120 TI - Characteristics of maltreatment experience and academic functioning among maltreated children. AB - This article examined the impact of maltreatment characteristics on academic functioning in a sample of school-age maltreated children. Results revealed several differences among three types of maltreatment subgroups (physical abuse, neglect, and sexual abuse) on characteristics of maltreatment experience. Few maltreatment characteristics were significantly related to measures of academic functioning, but the findings suggest that the effects of maltreatment characteristics should be examined separately for different types of maltreatment. PMID- 11437121 TI - High school students' responses to dating aggression. AB - The purpose of this study was to identify high school students' actions in response to physical aggression in their dating relationships. The association of these actions with race/ethnicity and gender was also examined. From a sample of high school students (N = 476), a subsample who reported that they had experienced at least one episode of being victimized by physical aggression in a dating relationship (n = 183), served as the sample of interest. On average, students engaged in two help-seeking actions, with females reporting more actions than males. Overall, the most common responses to physical aggression in a dating relationship were aggressive action (e.g., fight back), informal help seeking, threatened or actual breakup, and doing nothing (males) or crying (females). Females were more likely to fight back than were males. Race was largely unrelated to students' actions. Intervention opportunities and areas for future research are discussed. PMID- 11437122 TI - New thiadiazine derivatives with activity against Trypanosoma cruzi amastigotes. AB - The cytotoxicity of 18 new 1,2,6-thiadiazin-3,5-dione 1,1-dioxides was evaluated. This group of products was previously assayed against epimastigotes of Trypanosoma cruzi and some of them showed a high antiprotozoal activity. Thereafter 13 compounds with a high anti-epimastigote activity and low cytotoxicity were selected to be assayed against amastigotes. Some of the products showed the same or even lower cytotoxicity than nifurtimox and benznidazole, but most of them were very toxic for macrophages at 100 microg/ml. Only one of the compounds had an anti-amastigote activity similar to that of reference drugs at 10 microg/ml, but unfortunately this disappeared at lower concentrations. PMID- 11437123 TI - Weketrema gen. n., a new genus for Weketrema hawaiiense (Yamaguti, 1970) comb. n. (Digenea: Lecithasteridae) recently found in Australian marine fishes. AB - A new genus, Weketrema, is erected in the family Lecithasteridae for the species hitherto known as Lecithophyllum hawaiiense. Weketrema hawaiiense (Yamaguti, 1970) comb. n. is redescribed from Scolopsis bilineatus (Bloch) (Perciformes: Nemipteridae) from Lizard Island and Heron Island, Queensland, Plectorhinchus gibbosus (Lacepede) (Perciformes: Haemulidae) from Heron Island and Cheilodactylus nigripes Richardson (Perciformes: Cheilodactylidae) and Latridopsis forsteri (Castelnau) (Perciformes: Latridae) from Stanley, northern Tasmania. The new genus is distinguished from related members of the family Lecithasteridae by its complete lack of a sinus-sac. Although placed in the subfamily Lecithasterinae pro tem, its true subfamily position is not entirely clear. Comment is made on its unusual distribution, both in terms of zoogeography and hosts. PMID- 11437124 TI - Psilolintonum lineatum (Linton, 1928) (Echinostomatoidea: Psilostomidae) re allocated to Podocotyle (Allocreadioidea: Opecoelidae) on the basis of a re examination of the type material. AB - The type-material of Psilostomum lineatum Linton, 1928 was re-examined and identified as Podocotyle reflexa (Creplin, 1825). This re-allocation of the type and only speciesinvalidates the genus Psilolintonum that is now considered a synonym of Podocotyle. PMID- 11437125 TI - Experimental infection of Atlantic salmon (Salmo salar) with marine Eubothrium sp. (Cestoda: Pseudophyllidea): observations on the life cycle, aspects of development and growth of the parasite. AB - The life cycle of marine Eubothrium sp. (Cestoda: Pseudophyllidea), from Atlantic salmon (Salmo salar L.) was experimentally completed in one year and included only one intermediate host (Acartia tonsa Dana) (Copepoda: Calanoida). Adult cestodes were collected from farmed salmon, and ripe eggs released by the cestodes were fed to Acartia tonsa. Ingested eggs hatched in the gut and the larvae developed in the haemocoel of the copepod for 15 days at 16 degrees C. A total of 170 seawater-reared salmon were exposed to infected copepods and the total prevalence of Eubothrium sp. in the salmon after infection was 95.3%, with a mean intensity of 15.0 (range 1-87). The infected salmon were kept in the laboratory where the growth of the cestodes was studied for eleven months. Mean length of the cestodes increased with time, but a large variation among the cestodes was observed. Growth and maturation of the cestodes were dependent on host size and the number of worms present in the intestine. No evidence of mortality of Eubothrium sp. was observed during the experimental period. PMID- 11437126 TI - Philometroides africanus sp. n. (Nematoda: Philometridae), a new tissue parasite of the African pike Hepsetus odoe (Pisces) in Botswana. AB - A new species of philometrid nematode, Philometroides africanus sp. n., is described from female specimens found encapsulated in gill arches and inner surface of gill covers of the African pike, Hepsetus odoe (Bloch), an endemic freshwater fish in Africa, from the Okavango River and Delta in Botswana. This new nematode is characterised mainly by a markedly small and plump body of gravid females (body length 6-9 mm), a separate anterior oesophageal bulb, a conspicuously small oesophageal gland, presence of four pairs of very small submedian cephalic papillae, and absence of any caudal processes. The prevalence of P. africanus in African pike of the Okavango Delta was 29%, with the intensity 1-8 (mean 3) encapsulated nematodes per fish. The genus Margolisianum Blaylock et Overstreet, 1999 is considered a junior synonym of Philometroides Yamaguti, 1935 and, consequently, its type species is transferred to the latter as Philometroides bulbosus (Blaylock et Overstreet, 1999) comb. n. PMID- 11437127 TI - Redescription of Mehdiella microstoma and description of Mehdiella petterae sp. n., with a new definition of the genus Mehdiella Seurat, 1918 (Nematoda: Pharyngodonidae). AB - The generic diagnosis of Mehdiella Seurat, 1918 is emended based on study and redescription of Mehdiella microstoma (Drasche, 1884) from the caecum of Testudo graeca Linnaeus, 1758 collected in Settat, Morocco and on study and description of a new species, Mehdiella petterae sp. n., from the large intestine of Testudo hermanni (Gmelin, 1789) collected in Catalonia, Spain. Scanning electron microscopy (SEM) studies revealed substantial differences in the structure of the mouth and the caudal end, and made possible to differentiate the new species from the others. SEM studies showed the real and sound characteristics of the genus Mehdiella, namely number of anal papillae 2 instead of 3, post-anal papillae pedunculate or sessile instead sessile. PMID- 11437128 TI - Redescription of Ichthyofilaria argentinensis Incorvaia, 1999 (Nematoda: Philometridae) parasite of Merluccius hubbsi (Pisces: Merlucciidae) from Argentina. AB - Ichthyofilaria argentinensis Incorvaia, 1999 (Nematoda: Philometridae) is redescribed from type specimens and from parasites collected from the swim bladder of Argentine hakes Merluccius hubbsi Marini, 1933 caught at the Gulf of San Jorge (46 degrees 30'S, 66 degrees 30'W), Argentine Sea, in July, 1998. Sixteen out of 80 hakes (20%) were found to be parasitised by gravid females of this philometrid. After revision of both these nematodes and type material, the following differences from the original description of the species were observed: (a) cuticle with fine transverse striations, instead of smooth cuticle; (b) presence of cephalic papillae and a pair of amphids; (c) absence of swellings in the oesophagus; (d) oesophageal gland appendix very large and lobulate rather than small and vermiform, and (d) presence of vulva and vagina. The morphological features given in the redescription allow the confirmation of the identity of I. argentinensis as a valid species and to distinguish it from other members of the genus. PMID- 11437129 TI - Composition and structure of helminth communities in two populations of Pipistrellus pipistrellus (Chiroptera: Vespertilionidae) from Spain. AB - The community composition and structure of helminths of Pipistrellus pipistrellus (Schreber, 1774) from two widely separated Spanish localities, El Saler (n = 42) and the San Pedro pothole (n = 34), were determined and compared. Five species of trematodes, Plagiorchis (Plagiorchis) sp., Lecithodendrium (Lecithodendrium) linstowi Dollfus, 1931, Prosthodendrium (Prosthodendrium) sp., Pycnoporus heteroporus (Dujardin, 1845) and Parabascus semisquamosus (Braun, 1900), and one species of cestode, Hymenolepis pipistrelli Lopez-Neyra, 1941, were found. The two bat populations harboured the same helminth species and showed the same trematode dominance, but the most important differences between the two helminth community structures were attributable to L. (L.) linstowi and H. pipistrelli. The mean species richness in the two localities was not significantly different. The mean number of helminth species per infected bat, mean infracommunity abundance and mean infracommunity diversity showed significant differences between both localities. The number of helminths per bat in both populations displayed an aggregated distribution. Results indicate that the different characteristics of the P. pipistrellus foraging area in both localities are important in determining the composition and structure of helminth communities in this bat species. This is the first study of a Palaearctic bat helminth community. PMID- 11437130 TI - Mortality caused by experimental infection with the yeast Candida haemulonii in the adults of Ornithodoros moubata (Acarina: Argasidae). AB - A relatively high rate of mortality among engorged females of Ornithodoros moubata (Murray, 1877) was observed in our laboratory colony. The general aim of the study was to identify the causative agent responsible for this mortality. The diagnostic tests were performed by Yeast Identification Service (CBS-Delft, Netherlands) and the pathogen was identified as the yeast Candida haemulonii (van Uden et Kolipinski, 1962) Meyer et Yarrovi, 1978. The artificial infection study was performed by intrahaemocoelic inoculation of yeast suspension, resulting in a mortality of 37%. The maximum mortality of ticks infected per os by contaminated blood meal was 13%. Re-isolated yeast cells from haemolymph of dead and paralysed ticks were apparently identical with primary yeast cells, without loosing reproductive abilities. An occasional formation of elongated chains of yeast cells (pseudomycelium) was recorded. The majority of ticks infected in both experiments mentioned above survived and displayed no evident symptoms of the infection. The presence of yeast cells in the haemolymph of surviving ticks was not detected. The in vitro phagocytosis assay performed with FITC-labelled yeast cells showed that about 4% of tick haemocytes were phagocytically active against the pathogenic yeast cells. Thus phagocytosis seems to be a potent defence reaction against spreading and multiplying of the yeast C. haemulonii within the tick haemocoel. PMID- 11437131 TI - Picobia polonica sp. n. (Acari: Prostigmata: Syringophilidae), a new species of quill mite from the domestic hen, Gallus gallus domesticus (Aves: Phasianidae). AB - Female, physogastric female and male of Picobia polonica sp. n. collected from the domestic hen Gallus gallus domesticus (L.) in Poland are described and the phenomenon of physogastry in this and related species is briefly discussed. Picobia polonica is similar to Picobia khulkhshani (Kivganov et Sharafat, 1995). These two species can be distinguished by the following characters: P. polonica: chelicerae dentate, each with three minute teeth; setae g1 and pg2 subequal in length; ratio pg1 : pg2, 3: 1. P. khulkhshani: chelicerae edentate; setae g1 twice shorter than pg2; ratio pg1 : pg2, 1.6: 1. PMID- 11437132 TI - Chromosome analysis of Proteocephalus osculatus (Cestoda: Proteocephalidea). PMID- 11437133 TI - Evidence of tubulin in the scolex gland ducts of Gymnorhynchus gigas plerocercoid (Cestoda: Trypanorhyncha). PMID- 11437134 TI - Diversity of gyrodactylids from some marine fishes in tropical and subtropical Queensland, Australia. PMID- 11437135 TI - The interrelationships of metazoan parasites: a review of phylum-and higher-level hypotheses from recent morphological and molecular phylogenetic analyses. AB - Phylogeny of seven groups of metazoan parasitic groups is reviewed, based on both morphological and molecular data. The Myxozoa (=Malacosporea + Myxosporea) are most probably related to the egg-parasitic cnidarian Polypodium (Hydrozoa?: Polypodiozoa); the other phylogenetic hypotheses are discussed and the possible non-monophyly of the Cnidaria (with the Polypodiozoa-Myxozoa clade closest to the Triploblastica) is suggested. The Mesozoa is a monophyletic group, possibly closely related to the (monophyletic) Acoelomorpha; whether the Acoelomorpha and Mesozoa represent the basalmost triploblast clade(s) or a derived platyhelminth subclade may depend on rooting the tree of the Triploblastica. Position of the monophyletic Neodermata (=Trematoda + Cercomeromorpha) within the rhabditophoran flatworms is discussed, with two major alternative hypotheses about the neodermatan sister-group relationships (viz., the "neoophoran" and "revertospermatan"). The Myzostomida are not annelids but belong among the Platyzoa, possibly to the clade of animals with anterior sperm flagella (=Prosomastigozoa). The Acanthocephala represent derived syndermates ("rotifers"), possibly related to Seison (the name Pararotatoria comb. n. is proposed for Seisonida + Acanthocephala). The crustacean origin of the Pentastomida based on spermatological and molecular evidence (Pentastomida + Branchiura = Ichthyostraca) is confronted with palaeontological views favouring the pre-arthropod derivation of the pentastomids. Phylogenetic position of the nematodes within the Ecdysozoa and evolution of nematode parasitism are discussed, and the lack of relevant information about the enigmatic ectoproctan parasite Buddenbrockia is emphasised. PMID- 11437136 TI - Pediatric hand fractures: a review. AB - OBJECTIVE: To identify the incidence, causes, and details of hand fractures in children. DESIGN: A retrospective chart review. METHODS: Records of children under 16 years of age who had sustained a hand fracture within the last 5 years were collected from the patient population of British Columbia's Children's Hospital. A total of 242 hand fractures in 232 patients were documented. These patients were reviewed for age at time of injury, gender, location of the incident, mechanism of injury, number of radiographs taken, and fracture specifics. Radiographs with obscure details or incomplete folders were excluded. RESULTS: The patients consisted of 57 (24.6%) females and 175 (75.4%) males, with a mean age of 11.1 +/- 3.3 years. Incidence was low in early childhood but rose sharply after age 9 and peaked at 12 years of age. Sporting activities were the most common cause in both sexes. The fifth metacarpal was the most frequently involved bone (21.1% of total). Nonepiphyseal fractures accounted for 60.2% of the fractures, and the remaining 39.8% were epiphyseal fractures, predominantly Salter-Harris type II (90.4%). Fractures with comminution, severe displacement, intraarticular involvement, and condylar involvement were seen in 12.4%, 12.4%, 20.5%, and 15.1%, respectively. An average of 4.2 radiographs were taken per patient. CONCLUSION: Almost all fractures healed in 2 to 3 weeks with excellent functional outcome. Knowledge of epidemiology and etiology of hand fractures can serve as an essential first step in devising strategies to reduce the incidence of these hand injuries. It is hoped that studies such as the present study may serve as a first step in planning measures to reduce the occurrence of hand fractures. PMID- 11437137 TI - Facial fractures in children. AB - OBJECTIVE: To determine the pattern of injury of facial fractures in children, the relative contribution of plain radiography and CT scanning in the diagnosis of these injuries, and factors leading to delayed diagnosis. DESIGN: Retrospective case note review. PARTICIPANTS: All children with facial fractures identified using the trauma and medical record databases at our institution. RESULTS: Forty-six children with 59 facial fractures presented over a 4-year, 2 month period from November 1995 to December 1999. The median age was 10 years, with a range from 1 to 14. There was a 2-to-1 male-to-female sex ratio. A motor vehicle accident (MVA) involving a child as passenger, pedestrian, or cyclist accounted for 63% of cases. In seven of these, the child was either a front seat passenger or inappropriately restrained for their age and size. In all but one case, the presence of a fracture was associated with an overlying laceration, abrasion, or significant soft tissue edema. Initial examination and plain radiologic assessment by a pediatric clinician led to diagnostic delay in nine children. Facial CT was performed in 38 children, and all results were positive. Twenty-six patients required operative intervention for their facial fracture. Associated injuries, particularly of the head and limbs, were present in all but six patients. CONCLUSIONS: Facial fractures were uncommon overall but occurred more frequently in children with major trauma. Plain facial radiographs provided limited additional diagnostic information to careful clinical examination and often fail to detect or clearly define a facial fracture in children. In the correct clinical setting, a facial CT scan allows accurate diagnosis of the injury and can reveal previously unsuspected additional fractures. PMID- 11437138 TI - Signs of meningeal irritation at the emergency department: how often bacterial meningitis? AB - OBJECTIVE: Although signs of meningeal irritation are highly indicative of meningitis, they are not pathognomonic. In this study, we described the final diagnoses in children with signs of meningeal irritation, and we assessed the frequency of bacterial meningitis related to specific signs of meningeal irritation. METHODS: Information was collected from records of 326 patients (aged 1 month to 15 years) who visited the emergency department of the Sophia Children's Hospital between 1988 and 1998 with signs of meningeal irritation, assessed by either the general practitioner or the pediatrician. RESULTS: Bacterial meningitis was diagnosed in 99 patients (30%), viral or aseptic meningitis in 43 (13%). Other diagnoses were pneumonia (8%), other serious bacterial infections (2%), and upper respiratory tract infections or other self limiting diseases (46 %). Presence of one of the signs of meningeal irritation assessed by the pediatrician was related to bacterial meningitis in 39%. Specific tests eliciting meningeal irritation, such as Brudzinski's and Kernig's signs, were not related to a higher frequency of bacterial meningitis than neck stiffness and the tripod phenomenon. In children < or =1 year, bacterial meningitis is more frequently related to presence of irritability and a bulging fontanel. CONCLUSION: Bacterial meningitis is present in 30% of children with signs of meningeal irritation. Presence of meningeal irritation as assessed by the pediatrician is related to bacterial meningitis in 39%. A better prediction of bacterial meningitis was not achieved by using more specific tests for signs of meningeal irritation. PMID- 11437139 TI - Complementary and alternative medicine use in children. AB - OBJECTIVE: Adults frequently use complementary and alternative medicines (CAM). Few studies have reported how often CAM therapies are used to treat children. The purpose of this study is to describe the use of alternative therapies by children visiting an emergency department (ED) and to identify sociodemographic factors that may influence the decision to use such therapies. DESIGN: Survey of families using a self-administered questionnaire. SETTING: An urban, tertiary care pediatric emergency department. PARTICIPANTS: Convenience sample of families presenting to the ED for acute care. MAIN OUTCOME MEASURE: Use of CAM therapies to treat children. Secondary measures include the type of therapies used, types of medical problems the therapies were used for, reasons for using such therapies, use of such therapies by the child's caretakers, and sociodemographic characteristics of the children and families. RESULTS: A review of 525 completed surveys identified 63 caretakers (12%) who acknowledged that they had used at least one form of CAM therapy to treat any of their children. Homeopathic and naturopathic remedies were the most common therapies used. Parents most often used CAM therapies to treat respiratory problems in their children and were most influenced by word-of-mouth. Children who were treated with CAM therapies were more likely to have a caretaker who used such therapies. Twelve (40%) of 30 families who reported using either an herbal or homeopathic remedy, also used a prescription or over-the-counter medication at the same time to treat their child. Thirty-nine of 55 families (70.9%) reported informing their child's physician of their use of CAM therapies. CONCLUSION: CAM therapies are frequently used to treat children. Most parents who use CAM therapies to treat their children use such therapies themselves. Large proportions of children who are taking herbal supplements are also taking prescription or over-the-counter medications concurrently. PMID- 11437140 TI - Ability of hospitals to care for pediatric emergency patients. AB - CONTEXT: The needs of children in emergency situations differ from those of adults and require special attention, yet there has been no study of the ability of U.S. hospitals to care for emergently or critically ill children. OBJECTIVE: To estimate the distribution of pediatric services available at U.S. hospitals with emergency departments (EDs). DESIGN: Self-report survey of 101 hospital EDs. PARTICIPANTS: Stratified probability sample of all U.S. hospitals operating EDs. RESULTS: The majority of hospitals that usually admit pediatric patients do not have separate pediatric facilities. Hospitals without a pediatric department, ward, or trauma service usually transfer critically injured pediatric trauma patients; however, nearly 10% of hospitals without pediatric intensive care facilities admit critically injured children to their own facilities. Likewise, 7% of hospitals routinely admit pediatric patients known to require intensive care to their adult intensive care units rather than transferring the patient to a facility with pediatric intensive care facilities. Few hospitals have protocols for obtaining pediatric consultation on pediatric emergencies. Appropriately sized equipment for successful care of infants and children in an emergency situation was more likely to be missing than adult-sized equipment, and significant numbers of hospitals did not have adequate equipment to care for newborn emergencies. CONCLUSION: Emergent and critical care of infants and children may not be well integrated and regionalized within our health care system, suggesting that there is room for improvement in the quality of care for children encountering emergent illness and trauma. PMID- 11437141 TI - Severe hypernatremic dehydration and death in a breast-fed infant. AB - Breast milk is acknowledged as the best source of nutrition for neonates. We present the case of a full-term newborn who was fed solely breast milk and developed severe dehydration and hypernatremia. The patient developed cerebral edema, transverse sinus thrombosis, and died. The literature on the uncommon entity of breast-feeding hypernatremia and dehydration is reviewed, and management strategies are presented. PMID- 11437142 TI - Bilateral calf pain in an adolescent. PMID- 11437143 TI - Acute hemorrhagic complication of diagnostic lumbar puncture. AB - OBJECTIVE: To present a case of an epidural hematoma after lumbar puncture in a pediatric patient without known risk factors for such a complication and to review the literature regarding this complication. DESIGN: Case report, review of the literature, and discussion. DATA SOURCES: A review of MEDLINE (1966-1998) for keywords "lumbar puncture" and "hemorrhage" or "hematoma" was conducted, and each bibliography was reviewed for other sources extending to 1911. Articles describing a case of spinal hematoma after a lumbar puncture for any procedure were included. RESULTS: A 5-year-old boy underwent a lumbar puncture for evaluation of lethargy and fever, and subsequently developed marked back pain and severe pain on flexion of his legs. Magnetic resonance imaging revealed an epidural blood collection. The patient's symptoms resolved over the next few days in association with steroid administration. Multiple reports of epidural and subdural hematomas were found on literature review, most occurring in the setting of coagulation abnormalities. These reports involve lumbar puncture in anesthetic, interventional, and diagnostic settings. CONCLUSION: Lumbar puncture is a frequently employed procedure. Known complications include epidural, subdural, and subarachnoid hemorrhage, usually in the setting of abnormal coagulation. The case presented is unusual in that the patient is a child and lacks any known risk factors for a hemorrhagic complication. Such a complication appears to be rare; only five of the 64 cases discovered in the literature review occurred following this diagnostic procedure in patients without known risk factors. PMID- 11437144 TI - Subclinical mastitis presenting as acute, unexplained, excessive crying in an afebrile 31-day-old female. AB - We present a case of a 31-day-old female who presented with an acute, unexplained, episode of excessive crying. The patient had no history of fever and no fever on presentation. There was no evidence of skin erythema or swelling on presentation. Chest radiograph and computed tomography of the head were normal. Urinalysis and cerebral spinal fluid analysis were normal. The white blood cell count was within normal limits (17,400 cells/mm3). It was not until 6 hours after the onset of inconsolable crying (2.5 hours after presentation to the emergency department) that the patient's temperature rose to 38.5 degrees C (101.3 degrees F). Clinical signs of mastitis did not become appreciable until the 2nd hospital day. We review the literature on infantile mastitis and acute, unexplained, excessive crying. The importance of a thorough, conservative approach to the infant with acute, unexplained, excessive crying is discussed. PMID- 11437145 TI - Cold-syrup induced movement disorder. PMID- 11437146 TI - The importance of serial neurologic examination and repeat cranial tomography in acute evolving epidural hematoma. AB - Computed tomography (CT) has revolutionized the diagnosis and management of head injured patients, and its increasing availability has led to its liberal use. CT scanning provides excellent anatomic detail of the brain as fixed static images, but the dynamic nature of human physiology means that many injury patterns will evolve in time. We describe an 8-year-old child who had fallen 8 feet from a tree. He had a brief loss of consciousness but a normal neurologic evaluation on arrival to the emergency department (ED). He underwent expedited cranial CT scanning, which revealed no acute brain injury. Two and one half hours later, the patient had a mild depression in consciousness, prompting a second CT scan in the ED, which revealed an acute epidural hematoma. He had acute surgical evacuation of the hematoma and made a full neurologic recovery. This case illustrates that a single early CT examination may at times provide a false sense of security and underscores the importance of serial neurologic examinations. PMID- 11437147 TI - Benign occipital epilepsy mimicking a catastrophic intracranial event. AB - OBJECTIVE: To describe the rare, dramatic, presentation of benign occipital epilepsy. METHODS: We describe three children who presented to the pediatric emergency department from 1992 to 1996 with a clinical picture of catastrophic intracranial event. RESULTS: The main signs and symptoms were loss of consciousness in all patients, apnea in two, hemiclonus in two, general hypertonicity in two, eye deviation in two, fixed dilated pupils in one, and decorticate rigidity in two. All underwent emergency intubation, brain scan, and lumbar puncture, and all were treated with antibiotics, in addition to antiviral drugs in two. Two patients were also treated for suspected increased intracranial pressure. Two patients recovered within a few hours and one within 24 hours of admission without any residual neurologic deficit. Electroencephalograms, done within 48 hours after the event, revealed the classic pattern of occipital epilepsy in two patients and bilateral occipital slow wave in one. A 3- to 5-year clinical and electroencephalographic follow-up supported the diagnosis. CONCLUSION: Benign occipital epilepsy in children can mimic a catastrophic intracranial event. Electroencephalography, performed early in the Pediatric Intensive Care Unit, may avoid or shorten unnecessary and aggressive treatments such as hyperventilation, diuretic agents, and prolonged antiviral therapy. PMID- 11437148 TI - Direct ophthalmoscopy in pediatric emergency care. AB - Thorough knowledge about the basic ophthalmic examination is instrumental in the emergency department (ED) to determine the severity of eye emergencies and to develop a plan for treatment, including the possible need for an ophthalmology consultation. The direct ophthalmoscope is undoubtedly the most practical instrument available to a pediatric emergency physician for ophthalmic examination. However, the information elicited by its use is dependent on the quality of the instrument and the competency of the user. A greater understanding of the range of possibilities of the direct ophthalmoscope as a diagnostic tool will facilitate more efficient care of the pediatric patient. PMID- 11437149 TI - Managerial dilemmas. PMID- 11437150 TI - Yet two more uses for Colace. PMID- 11437151 TI - Abdominal pain: motor vehicle accident. PMID- 11437152 TI - Pediatric emergency medicine: legal briefs. PMID- 11437153 TI - Emergency nursing care of pediatric sickle cell patients: meeting the challenge. AB - Emergency care of the pediatric sickle cell patient requires complex nursing care and interventions including patient assessment, pain management, infection control, and appropriate understanding of complex hematological and immunological issues. This article includes two case studies that will illustrate the pathophysiology of sickle cell disease in the context of the bedside emergency nursing of the pediatric patient. PMID- 11437154 TI - Novel psychotropic medications in children: new toxicities to master. PMID- 11437155 TI - New therapies for Parkinson's disease. AB - In the last decade there has been a surge of new therapeutic strategies for the treatment of Parkinson's disease along with a change of concepts about how the disease should be treated. The gold standard remains levodopa preparations, which have a rapid and dramatic symptomatic effect by replenishing the reduced dopamine levels in caudate and putamen nuclei. However, keeping in mind the complications that may emerge following long-term treatment, its initiation should possibly be delayed to the more advanced stages of the illness, especially in younger patients, in favour of dopamine agonists monotherapy. The adverse reactions that become prominent and disabling in late stages of the disease, i. e., dyskinesias, response fluctuations, and psychiatric side effects, can currently be managed by novel pharmacological as well as surgical strategies. Future therapies will focus on transplantation of dopaminergic embryonic tissue, gene therapy, and neuroprotective treatments. PMID- 11437156 TI - Neurosarcoidosis: a review of its intracranial manifestation. AB - Sarcoidosis is a multisystem disease of unknown cause and with a worldwide distribution. Involvement of the central nervous system occurs in a relatively small number of patients with sarcoidosis. Isolated neurosarcoidosis without signs of systemic disease is a rarity. Because of its non-specific clinical presentation and neuroradiological imaging characteristics, intracranial neurosarcoidosis remains a very difficult diagnosis, particularly in the absence of systemic signs of the disease. Intracranial neurosarcoidosis has a predilection for the basal leptomeninges commonly affecting the cranial nerves, but any part of the brain may be involved, resulting in a wide spectrum of clinical syndromes. Cranial nerve involvement is the most common single symptom. Intracranial sarcoid manifests as nodular or diffuse leptomeningeal thickening and extraor intra-axial parenchymatous lesions. Intracranial sarcoid may mimic various forms of meningitis, including carcinomatous and intracranial mass lesions such as meningioma, lymphoma and glioma, based on neuroradiological imaging. Magnetic resonance imaging is a very sensitive diagnostic tool for detecting intracranial abnormalities due to neurosarcoidosis. Lumbar puncture is useful in ruling out other neurological disorders, in particular infectious, but cerebrospinal fluid findings are not specific. Angiotensin-converting enzyme in serum and cerebrospinal fluid may be increased, decreased or normal. Therapy consists of immunosuppressive agents and should be initiated with corticosteroids. Other immunosuppressive drugs should be added in severe cases or after frequent recurrences. We review here all aspects of intracranial neurosarcoidosis from the clinical point of view, with special emphasis on presentation, diagnostic procedures, differential diagnostic considerations and treatment. PMID- 11437157 TI - Recurrent stroke and vascular events in elderly patients with anticardiolipin antibodies: a prospective study. AB - The presence of anticardiolipin antibodies (aCL) is a recognized risk factor for ischaemic stroke and a predictor of recurrent ischaemic events in young patients, but the significance of positive aCL tests is uncertain in the elderly. We evaluated the frequency of aCL and the risk of recurrence of stroke and other vascular events in a series of 242 consecutive patients aged over 60 years, admitted for brain infarction. All underwent aCL immunoreactivity (ELISA; measured by IgG antiphospholipid, GPL, units) and transoesophageal echocardiography and were later examined or contacted by telephone (mean 2.33 +/- 1.25 years, max. 4). Fifty patients (21 %) had at least l0 GPL units aCL. There were no differences between these and the other patients in the results of transoesophageal echocardiography, including mitral or aortic valvular thickening, atrial thrombus, atrial spontaneous contrast, strands, and aortic plaques thickness. None had IgG higher than 80 GPL units or was positive for anti beta2 glycoprotein I. Patients with at least 10 GPL units more often had a past history of cerebral infarction than patients lower aCL level. However, the incidence of recurrent stroke was 4.5 per 100 person-year in patients with more than 10 GPL units, and 2.7 per 100 person-year in those with more than 10 GPL units. Kaplan-Meier analysis for any vascular events showed no differences between the two groups. In contrast to young patients, elderly patients with 10 or more GPL units aCL and negative for anti-beta2 glycoprotein I do not seem to have a higher risk of vascular events. PMID- 11437158 TI - Thalamic stimulation for primary writing tremor. AB - We report a patient with primary writing tremor whose tremor was treated with thalamic stimulation. He had undergone trials of multiple oral medications with no benefit for his tremor. An electrode lead was implanted in the thalamic nucleus ventralis intermedius with nearly complete control of his tremor and no postoperative complications. We conclude that nucleus ventralis intermedius thalamic stimulation is safe and effective for primary writing tremor. PMID- 11437159 TI - Study of binding and neutralising antibodies to interferon-beta in two groups of relapsing-remitting multiple sclerosis patients. AB - Interferon (IFN)-beta is generally considered an effective treatment for multiple sclerosis (MS); however, some patients do not respond to this therapy, possibly due to the production of neutralising antibodies (NAB) which can prevent the biological effect of IFN-beta. We compared the two types of IFN-beta, the glycosylated IFN-beta1a and the non-glycosylated IFN-beta1b, as their chemical differences may entail differing immunogenic capacities. We studied 22 relapsing remitting MS patients treated with IFN-beta1a and 31 treated with IFN-beta1b for 1 year, using the same assay and criteria, to compare the two types of IFN-beta in their ability to induce binding and neutralising antibodies and examined the correlation of the findings with the clinical data. Binding antibodies to IFN beta1a and IFN-beta1b were determined by enzyme-linked immunosorbent assay. A bioassay was used to detect and quantify the NABs to IFN-beta, measuring the capacity of NABs to block the antiviral resistance induced by IFNs. Binding antibodies were found in 32 % of those treated with IFN-beta1a and in 52 % of those treated with IFN-beta1b; NABs were found in 14% and 24 %, respectively. Both groups showed a significant decrease in relapse rate during the first year of treatment. These results demonstrate that the IFN-beta1b molecule is more immunogenic than the IFN-beta1a molecule. This may be due to the non glycosylated, chemical structure of the former, which can produce aggregates and enhance antibody production. No association was found between the presence of NABs and the clinical status of the patients. PMID- 11437160 TI - Smile-wink phenomenon: aggravated narrowing of palpebral fissure by smiling after lenticulocapsular stroke. AB - Five patients are described who developed unilateral narrowing of the palpebral fissure that was aggravated by smiling. They had ischaemic lesions involving mainly the contralateral lenticulocapsular area. This rare but distinct symptom may be an action induced focal eyelid dystonia, and could be described as'smile wink' phenomenon. PMID- 11437161 TI - Brain metastases from unknown primary tumour: a prospective study. AB - The best management of patients with brain metastases from an unknown primary tumour is still unclear, as data are scarce and studies are retrospective. We report 33 patients with biopsy-proven brain metastases from a primary tumour not found at the first investigations, who were treated by surgery and/or radiotherapy and followed with serial CT until death. Median survival time for all patients was 10 months and survival rates at 6 months, 1 year and 2 years were 76 %, 42 % and 15 % respectively. Patients with single brain metastasis treated by gross total resection and whole-brain radiotherapy (WBRT) had a median survival of 13 months with 76% alive at 6 months, 57 % at 1 year and 19% at 2 years. Patients with multiple brain metastases who underwent either WBRT alone or WBRT preceded by gross total resection of the symptomatic lesions had a poorer prognosis: median survival of 6-8 months with 50-100% alive at 6 months, 17-20% at 1 year and none alive at 2 years. In 85% of patients with a single brain metastasis a significant improvement in neurological functions was observed after surgical resection; among patients with multiple brain metastases a neurological improvement was observed in all patients who had a resection of symptomatic lesions and only in a half of patients who had WBRT alone. During the follow-up the primary tumour was found in 27/33 patients (82 %) and was located in the lung in 78%. Between 1987 and 1991 (with limited screening for the primary tumour in the follow-up) the unknown tumours were 6/15 (40%); in the more recent period (1992-1996) (CT-based screening for the primary tumour in the follow-up) no primary tumour remained unknown but overall survival has not significantly improved. The number of brain metastases was the only significant factor affecting survival after both univariate and multivariate analysis. This study suggests that, in patients with both single and multiple brain metastases from an undetected primary site when first studied, surgery and/or WBRT enable the control of the brain disease, partly because the systemic disease may be silent for a prolonged time. Only a few asymptomatic patients may benefit from an early detection and treatment of the primary tumour during the follow-up. PMID- 11437162 TI - Frontal lobe dysfunction in essential tremor: a preliminary study. AB - 27 patients with essential tremor (ET) were studied to determine the cognitive feature of this condition. 15 familial cases and 12 cases with a family history Parkinson's disease (PD) were identified. Performances on frontal lobe tasks of ET patients were compared with those of 15 patients with PD and 15 healthy control subjects. The ET patients showed significant impairments both in attentional and conceptual thinking tasks, similar to those observed in the PD group. Despite the nosographic independence of the two conditions, data showed that the frontal lobe feature of ET was similar to those of PD, thus possibly suggesting a common dysregulation of dopamine pathways. PMID- 11437163 TI - Severe metabolic abnormalities in the white matter of patients with vacuolating megalencephalic leukoencephalopathy with subcortical cysts. A proton MR spectroscopic imaging study. AB - Vacuolating megalencephalic leukoencephalopathy (VML) with subcortical cysts is a neurodegenerative disorder clinically characterized by megalencephaly with onset in the first year of life, progressive ataxia, spasticity and relatively spared cognitive function. Conventional MRI findings consist of diffusely abnormal cerebral white matter with subcortical cysts. Recent single-voxel proton MR spectroscopy studies have shown mild metabolic abnormalities in the white matter. We report here a combined proton MR imaging and MR spectroscopic imaging (1H MRSI) study on 2 new, unrelated patients with this rare disorder. 1H-MRSI examinations, which can provide simultaneously metabolic information from many different brain regions, showed inhomogeneous decreases in all normally detected metabolites with significant widespread decreases in the ratio of N acetylaspartate to creatine+phosphocreatine and concomitant small increases in lactate in the white matter of both hemispheres. Metabolic abnormalities were milder in the frontal white matter and more severe in the posterior white matter. The 1H-MRSI pattern of the gray matter was normal in both patients. In one patient, a subsequent 1H-MRSI examination (performed 3 years after the first) confirmed the presence of widespread decreases in the ratio of N-acetylaspartate to creatine+phosphocreatine in the white matter. We conclude that severe metabolic abnormalities can be found in the white matter of VML patients. This suggests that, despite the apparently mild clinical course, a severe neurodegenerative process may occur in the white matter of these patients. PMID- 11437164 TI - Mutation analysis in Chariot-Marie Tooth disease type 1: point mutations in the MPZ gene and the GJB1 gene cause comparable phenotypic heterogeneity. AB - Charcot-Marie-Tooth disease type 1 (CMT1) is a demyelinating peripheral neuropathy most commonly caused by a DNA duplication on chromosome 17p11.2 including the peripheral myelin protein 22 (PMP22). Point mutations in the myelin protein zero gene (MPZ) and gap junction protein, beta-1 gene (GJB1) are also found in association with CMT1 or the subclass of CMT type X (CMTX), respectively. Recently point mutations in these genes have been found in patients showing the axonal variant of CMT, CMT type 2 (CMT2). We here describe the clinical and electro-physiological findings caused by two novel and two recently described MPZ mutations and six GJB1 mutations. Different MPZ and GJB1 mutations were associated with different grades of severity in CMT1 and CMTX. The novel MPZ Glu141st op mutation was associated with the axonal CMT2. We conclude that the clinical and electrophysiological heterogeneity among CMT patients carrying point mutations in MPZ and GJB1 is similar. Thus for clinical purposes CMT1 and CMT2 patients should be screened for mutations in these two genes after duplication on chromosome 17p11.2 has been excluded as the disease causing mutation. PMID- 11437165 TI - Depression and anxiety in multiple sclerosis. A clinical and MRI study in 95 subjects. AB - The aim of the present study was to investigate the relationship between involvement of specific areas of the brain and the occurrence of depression and anxiety in patients with multiple sclerosis. We studied 95 patients (62 women and 33 men, mean age 39.5 years, SD 11.2) with definite MS, 97 patients (65 women and 32 men, mean age 40.7, SD 11.9) suffering from chronic rheumatoid diseases and 110 healthy subjects (71 women and 39 men, mean age 40.1, SD 12.7). The disability, the independence, the cognitive performances, the depressive and anxiety symptoms were assessed. The diagnosis of major depression was made according to the DSM-IV. The patients with multiple sclerosis underwent a 1.5 Tesla magnetic resonance examination including T1 and T2 weighted images. Calculation of regional and total lesion loads and brain volumes were performed. The number (%) of subjects with a diagnosis of major depression was 18 (18.9) among MS cases, 16 (16.5) among controls with chronic disease (p=NS), and 4 (3.6) among healthy volunteers (p < 0.0001). The Hamilton Depression and Anxiety rating scales median scores were 5 and 18, respectively in the MS patients, 5 (p= NS) and 14 (p= NS) in the chronic rheumatoid diseases controls, and 3 (p= < 0.0001) and 6 (p= < 0.0001) in the healthy controls. Both severity of depressive symptoms and diagnosis of major depression correlated, albeit weakly, with right frontal lesion load (r=0.22, p=0.035, and r=0.23, p=0.026, respectively) and right temporal brain volume (r=0.22, p=0.005 and r=0.22, p=0.036, respectively). The severity of depression was related significantly also with total temporal brain volume (r=0.26, p=0.012), right hemisphere brain volume (r=0.25, p=0.015), disability (r=0.30, p=0.003) and independence of MS cases (r=-0.26, p=0.01). The anxiety did not correlate significantly with any of the measures of regional and total lesion loads and brain volume or with any of the considered clinical variables. The similar frequency of depression and severity of depressive symptoms in MS patients and in chronic disease patients, the significant difference in this respect with the normal controls, and the significant correlation between depression and the disability measures would suggest a psychological reaction to the impact of the disease but the relationship between depression and the alterations in the frontal and temporal lobes of the right hemisphere supports, on the contrary, the causative role of organic brain damage. The lack of any significant association between symptoms of anxiety and either MRI abnormalities or clinical variables led us to the opinion that anxiety is a reactive response to the psychosocial pressure put on the patients. PMID- 11437166 TI - Writer's cramp induced by olanzapine. PMID- 11437167 TI - REM-associated hallucinations and sleep paralysis are dependent on body posture. PMID- 11437168 TI - Bilateral sciatic nerve "Saturday night palsy". PMID- 11437169 TI - Levodopa monotherapy can induce "sleep attacks" in Parkinson's disease patients. PMID- 11437170 TI - Clinical and magnetic resonance imaging evidence of brain and spinal cord demyelination in a case of chronic inflammatory demyelinating polyneuropathy. PMID- 11437171 TI - Guillain-Barre syndrome resembling brainstem death in a patient with brain injury. PMID- 11437172 TI - Tarsal tunnel syndrome caused by synovial sarcoma. PMID- 11437173 TI - A case of myasthenia gravis accompanied by erythema elevatum diutinum and rheumatoid arthritis. PMID- 11437174 TI - Pathophysiology of pre-eclampsia/eclampsia syndrome. PMID- 11437175 TI - Arthur Van Gehuchten (1861-1914). PMID- 11437176 TI - Plasma extracellular superoxide dismutase activity in healthy pregnant women is not influenced by zinc supplementation. AB - We hypothesized that plasma extracellular superoxide dismutase (EC-SOD) activity reflects the zinc nutriture of healthy pregnant women. Sixty-three women were selected from 580 African-American women who participated in a clinical trial to evaluate the effect of prenatal zinc supplementation on pregnancy outcome. Half of the women received zinc (25 mg/d) and the other half was given a placebo from about 19 wk gestation to delivery. In the trial, a positive effect of zinc supplementation on birthweight was observed, indicating that the population as a whole had suboptimal zinc nutriture. Using plasma samples obtained during the trial, EC-SOD activities were measured and the values were compared with plasma zinc concentrations and plasma alkaline phosphatase activities. Plasma EC-SOD activities in our subjects were lower than previously published values for healthy adults in Korea. Although plasma EC-SOD activity may reflect severe zinc deficiency, it is not a sensitive marker for marginal deficiency status. Plasma EC-SOD activities did not prove to be a better indicator of zinc nutriture of pregnant women than either plasma zinc or plasma alkaline phosphatase activities. PMID- 11437177 TI - Antioxidant status in cerebrovascular accident. AB - Ischemia is associated with the pathological changes caused by the accumulation of reactive oxygen metabolites (ROM) in cerebrovascular accident (CVA). The aim of this study was to determine red cell copper/zinc-superoxide dismutase (Cu/Zn SOD) and catalase activities and copper and zinc concentrations both in plasma and in red cells in CVA. Cu/Zn-SOD and catalase activities of 16 patients, with an average age of 64 yr, were measured spectrophotometrically; copper and zinc concentrations were determined by atomic absorption spectrophotometer. The results showed that Cu/Zn-SOD activity was increased markedly in patients compared to the young controls and reached a peak on the d 5 of the disease, whereas the catalase activity of the patients on d 3 and d 5 were in the normal range, but higher on d 10. The enzyme activities of the elderly group were generally increased compared to the young controls. Copper and zinc concentrations showed corresponding alterations. These findings suggested that the effects of oxidative stress in CVA might be reflected in red cell and plasma parameters. PMID- 11437178 TI - Dietary selenium intake of Chinese adult women in the 1990s. AB - To assess the levels of daily dietary intake of selenium (Se) among the general Chinese population, a series of field surveys were conducted in the 1990s. Samples of 24-h duplicates of foods were collected from 500 participants (300 in 6 cities and 200 from 4 villages). Se levels were determined by microwave digestion followed by inductively coupled plasma-mass spectrometry (ICP-MS), and the measurements were compared with FCT (Food Composition Tables)-based estimates. The average daily intake of Se was 69.2 lg/d (79.9 and 53.1 microg/d in urban and rural areas, respectively) by instrumental determination and 35.1 microg/d (36.7 and 32.7 microg/d) by FCT-based estimation. As the distribution of Se should be uneven within China, the FCT-based estimation is of a limited value and the ICP-MS determination of Se is more accurate and reliable when evaluating the nutritional status of local people. Taking ICP-MS-based values, Se intakes were lower in rural areas than in urban areas, and the intakes of about half of the people in rural areas were less than the Recommended Daily Allowance (RDA) in China of 50 microg/d. The low intake might be resulted from difference in the types of food consumed. Thus, the dietary intake of Se basically meets the recommended RDA in most of urban areas, but insufficiency may be still a nutritional and public health problem in some rural areas. PMID- 11437179 TI - Inhibitory effects of echinoisoflavanone and sophoraisoflavanone D in Sophora chrysophylla SEEM on lipid peroxidation of mice brain homogenate by interaction of ferrous ion and hydrogen peroxide, in vitro. AB - We present the results of an in vitro investigation of the inhibitory effects of echinoisoflavanone and sophoraisoflavanone D isolated from Sophora chrysophylla SEEM on lipid peroxidation of mice brain homogenate by interaction of ferrous ion and hydrogen peroxide, in vitro. They inhibited lipid peroxidation. The order of inhibitory effects of these isoflavanones and mannitol as a hydroxy radical scavenger was echinoisoflavanone > mannitol > sophoraisoflavanone D. The results suggest that some isoflavanones may be of use in cases where oxidative stress is present. PMID- 11437180 TI - Age-related changes in the concentrations of major and trace elements in the brain of rats and mice. AB - Age-related changes in the concentrations of constituent elements in the brains of rats and mice 1 wk to 24 mo old were determined with inductively coupled plasma-mass spectrometry (ICP-MS) and inductively coupled plasma-atomic emission spectrometry (ICP-AES). Seventeen elements could be determined with reasonable accuracy and reproducibility. They were P, K, Na, Mg, Ca, Fe, Zn, Cu, Rb, Al, Mn, Sr, Mo, Co, Pb, Cs, and Cd in order of concentrations in the adult rat brains. In these elements, six major elements (P, K, Na, Fe, Mg, Ca) were determined with ICP-AES and the others with ICP-MS. The concentrations of each element and the pattern of age-related changes were similar between the rat and mouse brains. The elements of which concentrations decreased with aging were K and Rb. On the other hand, the concentrations of some metal elements, including Fe, Cu, Sr, and Co, appeared to increase with growth and aging. The concentrations of other elements were relatively constant throughout the age examined. PMID- 11437181 TI - Effect of long-term treatment with vanadate in drinking water on KK mice with genetic non-insulin-dependent diabetes mellitus. AB - The glucose-lowering effect of vanadate, ammonium metavanadate (AMV), on diabetic KK mice was examined. Five-week-old male KK mice were administrated with a solution of AMV via drinking water at concentrations of vanadium (V) with 0.1, 1.0, 10 and 100 microg/mL for a period of 10 wk, respectively. Body weight, consumption of food and water, and blood glucose levels was measured every week for 10 wk. The results showed that food consumption and body weight in the experimental groups were similar to those in the control group. A statistically significant decrease of drinking water consumption and blood glucose levels in the group treated with 100 microg V/mL was observed. The glucose tolerance in the vanadate-treated mice with 10 and 100 microg V/mL was remarkably improved compared with the control group. Biochemical analyses at the end of experiments demonstrated that a distinct tendency for the glucose and hemoglobin A1c (HbA1c) levels to decrease with vanadate treatment in the blood was also observed. The glutamic pyruvic transaminase, glutamic oxaloacetate transaminase, blood urea nitrogen, triglyceride, high-density lipoprotein, and total cholesterol levels in plasma were lower in the higher vanadium groups than those in the control group. These results indicate that vanadium effectively produced the glucose-lowering effect at a higher dose than that at a low dose of vanadium in drinking water, without any overt signs of toxicity. PMID- 11437182 TI - Changes in zinc levels and superoxide dismutase activities in the skin of acute, ultraviolet-B-irradiated mice after treatment with ginkgo biloba extract. AB - Acute ultraviolet-B (UV-B) irradiation is known to act as an initiator in the formation of reactive oxygen species. These oxygen products are highly reactive and they are able to cause irreversible damage to cellular components. Oxygen free radicals are normally neutralized by very efficient systems in the body. These include antioxidant enzymes like superoxide dismutase (SOD). In a healthy subject, there is a balance between free radicals and the levels of antioxidants. In some pathological conditions such as oxidative stress, the level of antioxidants is significantly reduced. The skin contains relatively high levels of zinc (Zn), an essential element known to be a cofactor in some metabolic pathways. Zinc has also been reported to have antioxidant properties. In the present study, we investigated the effect of ginkgo biloba extract (Gbe), a potent free-radical scavenger, on UV-B-irradiated skin by measuring SOD activity and Zn levels in the skin, before and after treatment. The SOD activity was decreased after UV-B exposure, in comparison with the control group (p < 0.05). After Gbe treatment, the SOD activity increased (p < 0.05) as compared with the untreated UV-B irradiated group. The Zn levels changed in the same pattern as the SOD activity values. PMID- 11437183 TI - Effect of aluminum on the blood-brain barrier permeability in acute and chronically hyperglycemic rats. AB - This study examined the changes in blood-brain barrier (BBB) permeability following acute aluminum (Al) exposure during acute and chronic hyperglycemia in rats. Acute hyperglycemia was induced by intraperitoneal injection of glucose solution at 30 min after giving Al. Chronic hyperglycemia was made by an injection of alloxan monohydrate. BBB permeability was measured in the four regions of the brain at 1 h after administrating Al by spectrophotometric quantification of Evans blue (EB) dye. The extravasation of EB dye was significantly more extensive in the two regions of brain in the groups treated with Al, Al plus glucose, and alloxan plus Al than in the groups treated with saline, glucose, and alloxan alone (p < 0.05). Under acute and chronic hyperglycemia plus Al treatment, the BBB permeability to EB was significantly higher than that observed solely in Al-treated rats (p < 0.05). These data indicate that Al toxicity leads to an additional increase in BBB permeability, in which acute and chronic hyperglycemia potentiates the effects of Al to enhance BBB permeability to EB. PMID- 11437184 TI - Molybdenum supply of very low-birth-weight premature infants during the first months of life. AB - This explorative study was performed to assess basic data on the Mo metabolism of premature infants. Premature (n = 18, gestational age < or = 32 wk, birth weight < or = 1,500 g) and healthy formula-fed term infants (n = 14) were nourished and corrected for gestational age, identically. Plasma was collected at 3, 16, and 52 wk and 72 h balances were performed at 3 wk of age. In the premature infants, these investigations were preceded by two balance studies and an initial plasma collection. Increased Mo intake and low relative urinary excretion resulted in a retention of 4.4 (0.99-7.77) microg Mo/kg initially in premature infants (median, range). Parallel plasma concentrations were 5.5 (2.5-7.3) microg Mo/L, declining to 2.36 (0.73-3.87) microg Mo/L at 4 wk. Term infants rendered 1.49 (0.29-1.7) microg Mo/L (p < 0.05), with no significant differences later. It was concluded that the supplementation of formulas for premature infants with Mo should be recinded until there is evidence for its necessity. Index Entries: Mo; premature infants; trace elements; formula; nutrition. PMID- 11437185 TI - Pharmacological treatment of obesity in paediatric patients. AB - The prevalence and severity of obesity appear to be increasing in the paediatric population. This has resulted in an increasing trend in outcomes related to obesity, such as type 2 diabetes mellitus. The current clinical approach to management of obesity in paediatric patients is behavioural therapy directed at changing diet and physical activity. While there are no pharmacological agents currently approved for the treatment of paediatric obesity, there is hope that agents approved for use in adults will prove useful in adolescents and children. Therapeutic trials are currently under way to evaluate some agents, e.g. orlistat and sibutramine. Controlled clinical trials are necessary to evaluate drugs for weight management in children and adolescents. It should not be assumed that the risks and benefits associated with these drugs are the same for children as for adults. It is also possible that as the understanding of the science of obesity advances, new agents will be developed which will have appropriate benefits and risks for use in children. PMID- 11437186 TI - Treatment of status epilepticus in children. AB - Status epilepticus (SE) is a condition characterised by frequent and prolonged epileptic seizures which frequently develop in the immature brain. Fever, metabolic disorders and subtherapeutic concentrations of antiepileptic drugs are the most common factors precipitating SE in children. Progressive neuronal damage occurs if convulsive SE persists for more than 30 minutes, with neurological, epileptic and cognitive sequelae. Unfortunately, the immature brain is more predisposed to SE and its sequelae than the mature brain. SE may be categorised as convulsive, nonconvulsive or neonatal according to its responsiveness to antiepileptic drugs. Regardless of category, the main objective in the treatment of SE is to abort the seizures and treat the inciting condition. Treatment includes: (i) monitoring of hydration, electrolyte balance, and cardiocirculatory and pulmonary functions; and (ii) rapid intravenous administration of specific antiepileptic drugs. Benzodiazepines (usually diazepam, lorazepam or midazolam) are the most effective agents for the initial treatment of convulsive and nonconvulsive SE. In particular, midazolam infusion is an effective and well tolerated therapeutic approach for the management of childhood SE, including refractory SE. Phenytoin remains an excellent agent because of its long duration of action, but it is not active in nonconvulsive SE. Fosphenytoin, a phenytoin prodrug, represents a significant advance in the treatment of children with convulsive SE. Intravenous phenytoin and intramuscular phenobarbital (phenobarbitone) are generally used in neonatal SE; other agents are rarely used. PMID- 11437187 TI - Stroke in children with sickle cell anaemia: aetiology and treatment. AB - Cerebral infarction is a frequent, severe complication of sickle cell anaemia. During childhood, most strokes are due to infarction with the majority resulting from occlusion of the large cerebral arteries. Risk factors include transient ischaemic attacks, acute chest syndrome, severe anaemia and elevated blood pressure. Less certain is the association with leucocytosis, or protection provided by alpha-thalassaemia or fetal haemoglobin. Children who have one stroke are at significant risk for having subsequent events that can be substantially reduced by maintaining haemoglobin S below 30%. It has not yet been possible to identify individuals for whom transfusion can be safely stopped. Haemosiderosis is a consequence of intensive and long term transfusion therapy, which requires chelation with deferoxamine. Iron accumulation can be minimised using erythrocytapheresis but this is technically difficult in children, expensive and results in increased donor exposure. In addition to lesions associated with strokes, an additional 17% of patients can be shown to have clinically silent cerebral infarcts. Although these are termed 'silent', those affected have mild neuropsychological deficits. Their relationship to stroke or risk for recurrence is unknown. Transfusion therapy has been shown to provide primary stroke prevention for children who have elevated cerebral artery velocity. Finally, intracranial haemorrhages, more commonly found in adults, also affect children. Subarachnoid haemorrhage is frequently found to result from cerebral artery aneurysms. A condition that mimics the moyamoya syndrome radiographically, as well as for its risk of haemorrhage, can be found in children with partly occluded cerebral arteries either as a result of stroke or silent infarct. PMID- 11437188 TI - Rasburicase. AB - Rasburicase (recombinant urate oxidase) is a uricolytic agent which has been developed for the prevention and treatment of chemotherapy-induced hyperuricaemia at a dosage of 0.15 or 0.2 mg/kg/day in patients with haematological malignancies. Significant reductions from baseline in plasma uric acid levels were seen in 3 noncomparative and 1 randomised comparative trial of rasburicase in patients with, or at high risk of, hyperuricaemia. A multicentre, nonblind, randomised trial found that patients who received intravenous rasburicase 0.2 mg/kg/day experienced an average 2.6-fold less exposure to uric acid than those who received oral allopurinol 10 mg/kg/day. There was no evidence of renal insufficiency in 3 noncomparative trials which monitored creatinine levels to assess renal function during chemotherapy and rasburicase treatment. Rasburicase was generally well tolerated in clinical trials, with skin rashes reported in approximately equals 2% of patients. Bronchospasm, nausea and vomiting, and haemolysis occurred less frequently. In all cases the patients had also received chemotherapy prior to the adverse event. PMID- 11437190 TI - The effects of early motor intervention on children with Down syndrome or cerebral palsy: a field-based study. AB - This study reports the results of a field-based investigation of the effects of two motor intervention approaches--neurodevelopmental treatment and developmental skills--on children with two different diagnoses. The sample included 50 children, of whom 27 were diagnosed as having Down syndrome and 23 as having cerebral palsy. Children had a mean chronological age of 14 months at the beginning of the study. The severity of their motor impairment was assessed with the Gross Motor Function Classification System for Cerebral Palsy. Children's motor functioning was examined at entry into the study and after they received 1 year of motor intervention services. Dependent measures included children's general development, rate of motor development, and quality of movement. Pre-post comparisons indicated that children made significant changes in their motor development age and quality of movement over the course of intervention. However, there was no evidence that motor intervention accelerated development or improved quality of movement beyond what could be expected on the basis of maturation. Furthermore, no differential intervention effects were associated either with children's diagnosis or treatment model. Regression analyses indicated that the rate of motor development children attained after 1 year of intervention was highly related to their rate of development at the onset of intervention and, to a lesser degree, to the number of sessions of intervention children received. These results are discussed in terms of the need for the field of motor intervention to develop new treatment paradigms. PMID- 11437189 TI - Ondansetron: a review of its use as an antiemetic in children. AB - Ondansetron, a selective serotonin (5-hydroxytryptamine; 5-HT) 5-HT3 receptor antagonist, is an antiemetic agent available for use in adults and children. In children receiving ondansetron (multiple 5 mg/m2 or 0.15 mg/kg intravenous and/or oral doses) in addition to chemotherapy in 2 large (n > 100) non-comparative analyses, < or =2 emetic episodes were observed in 33 and 40% of cisplatin recipients, 48 and 68% of ifosfamide recipients, and 70 and 72% of patients receiving other chemotherapeutic regimens. In comparative trials, ondansetron was significantly more effective at reducing nausea and vomiting than metoclopramide or chlorpromazine (both combined with dexamethasone), although the incidence of delayed symptoms were similar between children receiving ondansetron and metoclopramide. In addition, dexamethasone significantly improved the antiemetic efficacy of ondansetron in 1 randomised trial. When used in children undergoing conditioning therapy (including total body irradiation) prior to bone marrow transplantation, ondansetron was significantly better at controlling nausea and vomiting than combined perphenazine and diphenhydramine therapy. In dose-ranging and large placebo-controlled trials, intravenous (0.075 to 0.15 mg/kg) or oral (0.1 mg/kg) ondansetron was significantly more effective than placebo in preventing emesis in children undergoing surgery associated with a high risk of postoperative nausea and vomiting (PONV) including tonsillectomy or strabismus repair. In comparative studies, intravenous administration of ondansetron 0.1 to 0.15 mg/kg was significantly superior to droperidol 0.02 to 0.075 mg/kg or metoclopramide 0.2 to 0.25 mg/kg in preventing emesis in children undergoing various surgical procedures. In comparison with other antiemetics, including prochlorperazine and dimenhydrinate, ondansetron generally showed greater prophylactic antiemetic efficacy. Ondansetron combined with dexamethasone was significantly more effective than ondansetron or dexamethasone alone, as was the combination of ondansetron with a propofol-based anaesthetic compared with either agent alone. Ondansetron is generally well tolerated in children, rarely necessitating treatment withdrawal. The most frequently reported adverse events were mild to moderate headache, constipation and diarrhoea in patients receiving chemotherapy. Wound problems, anxiety, headache, drowsiness and pyrexia were reported most frequently in patients postsurgery. CONCLUSIONS: Ondansetron has shown good efficacy in the prevention of acute nausea and vomiting in children receiving moderately or highly emetogenic chemotherapy and/or irradiation, particularly when combined with dexamethasone. In the chemotherapy setting, ondansetron is significantly better than metoclopramide and chlorpromazine and has a more favourable tolerability profile. In children undergoing surgery, ondansetron demonstrated superior prophylactic antiemetic efficacy compared with placebo, droperidol and metoclopramide, and was relatively free of adverse events. Ondansetron is thus an effective first-line antiemetic in children undergoing chemotherapy, radiotherapy and surgery. PMID- 11437191 TI - Can teachers' global ratings identify children with academic problems? AB - Physicians often elicit ratings from teachers when making diagnostic, treatment, or referral decisions. The purpose of this study was to view the relationship between teachers' ratings and children's academic skills, assess the utility of teacher ratings in detecting academic problems, and thus determine whether physicians can depend on teacher ratings when making decisions about patients' needs. Subjects were a national sample of 80 teachers and 934 children between 6 and 13 years of age participating in a test standardization study. Families were representative of United States demographics in terms of parental level of education, income, and ethnicity, and sites were geographically diverse elementary schools. Children were administered the Comprehensive Inventory of Basic Skills--Revised (CIBS-R), a diagnostic academic achievement test. Teachers rated children's academic performance on a five-point scale ranging from far above average to far below average and were blinded to the results of the CIBS-R. Teacher ratings varied significantly with children's performance for all academic domains. Logistic regression revealed that teacher ratings were best predicted by children's performance in basic reading skills, followed by math skills, and were not influenced by race, parents' level of education, history of retention, or gender. Participation in Title I services, testing in winter or spring, and parents who spoke a language other than English produced significantly lower ratings. Nevertheless, teachers rated as average many students with mild to moderate academic difficulties. School system personnel and health care providers should avoid sole dependence on global teacher ratings when deciding which students need special education referrals or other services. Supplementing teacher ratings with standardized screening test results is needed to ensure accurate decision-making. PMID- 11437192 TI - Situational analysis of parenting problems for caregivers of children with sickle cell syndromes. AB - A situational analysis of problematic situations was conducted for 37 caregivers of children with sickle cell disease (SCD) who ranged in age from 5 to 13 years. Participants responded to a semistructured interview related to caring for a child with SCD. The interview included the domains of medication adherence, nutrition, minimizing and coping with pain episodes, social problems, academic difficulties, and children's expression of negative feelings related to having SCD. Caregivers described a total of 356 problems. Almost all caregivers reported experiencing problems with their children's nutrition (n = 35), minimizing pain episodes (n = 34), and their children expressing feelings about having SCD (n = 33). Moderately challenging and emotionally upsetting problems were reported for coping with pain episodes. The total number of problems was significantly higher for boys than for girls. Nutrition issues were more frequently reported for younger children. Findings have salient clinical implications for the care of children with SCD. PMID- 11437193 TI - Effects of reducing television viewing on children's requests for toys: a randomized controlled trial. AB - Previous attempts to reduce the effects of television advertising on children's purchase requests have had little success. Therefore, we tested the effects of a classroom intervention to reduce television, videotape, and video game use on children's toy purchase requests, in a school-based randomized controlled trial. Third- and fourth-grade children (mean age, 8.9 years) in two sociodemographically and scholastically matched public elementary schools were eligible to participate. Children in one randomly selected elementary school received an 18-lesson, 6-month classroom curriculum to reduce television, videotape, and video game use. In both schools, in September (before intervention) and April (after intervention) of a single school year, children and parents reported children's prior week's purchase requests for toys seen on television. After intervention, children in the intervention school were significantly less likely to report toy purchase requests than children in the control school, with adjusting for baseline purchase requests, gender, and age (odds ratio, 0.29; 95% confidence interval, 0.12-0.69). Among intervention school children, reductions in self-reported purchase requests were also associated with reductions in television viewing. There was no significant difference between schools in parent reports of children's requests for toy purchases. These findings suggest that reducing television viewing is a promising approach to reducing the influences of advertising on children's behavior. PMID- 11437194 TI - Children and TV advertising: nowhere to run, nowhere to hide. PMID- 11437195 TI - A child with a learning disability: navigating school-based services. PMID- 11437196 TI - Presidential address. Society for Developmental and Behavioral Pediatrics. PMID- 11437197 TI - The effect of cooling on muscle co-ordination in spasticity: assessment with the repetitive movement test. AB - PURPOSE: Cooling muscles might produce a temporary reduction of spasticity. This study investigated muscle co-ordination in spasticity under the influence of cooling. METHODS: A repetitive movement (RM-) test of the ankle was used, while measuring the angle and surface-electromyography (EMG) of the m. tibialis anterior and m. triceps surae. Ensemble averaging provided quantified measures of muscle activation. Sixteen patients with spasticity in their lower extremity due to stroke or spinal cord injury participated in the study. Physical examination and the RM-test was done before and after cooling the m. triceps surae for 20 minutes by coldpacks. RESULTS: The results show that Achilles hyperreflexia and clonus were abolished in all, and all but one patient, respectively. The EMG of the m. triceps surae, acting as a prime mover, was increased (p = 0.028). However, this improved muscle co-ordination resulted in just a slightly increased active range of motion (less than 2 degrees at p = 0.049). CONCLUSION: Apparently, the increase in excitability of the alpha motoneuron pool in voluntary movements of patients with spasticity is not followed by an improvement in the ability to move. PMID- 11437198 TI - Classification of problems, clinical findings and treatment goals in patients with low back pain using the ICIDH-2 beta-2. AB - PURPOSE: This study aimed to explore problems, clinical findings and treatment goals presented by patients with low back pain and their physiotherapists, and to which extent this information could be classified according to the ICIDH-2 Beta 2. METHOD: An explorative field trial of 12 physiotherapists and 28 patients with low back pain in outpatient practices. RESULTS: Pain and impairments related to muscles, joints and structures of the lower back, pelvis and hips were most often presented. Functioning related to daily activities and participation in society was often described in the treatment goals, but vaguely formulated. Most of the problems and clinical findings could be classified according to the ICIDH-2 Beta 2. However, several treatment goals could not be classified. CONCLUSIONS: There was a lack of specification in several of the detailed codes of the Body, Function and Structure Dimension, particularly regarding classification of pain and muscle functions. The main limitation concerned the detailed version of the Body Function and Structure Dimension, which provided no additional precision compared with the short version when classifying pain and other impairments. However, the ICIDH-2 offers a tool to enhance the precision of clinical findings and the possibilities for performing a multidimensional evaluation of functioning related to patients with low back pain. PMID- 11437199 TI - Predicting return to work after brain injury using occupational therapy assessments. AB - PURPOSE: The aim of this study was to evaluate the value of occupational therapy assessments used in an outreach rehabilitation programme to predict return to work after brain injury. The assessments represent the ICIDH-2 levels of body function and activity. METHOD: Fifty-six persons in a late phase after brain injury who had been admitted to the rehabilitation programme during 2 years were followed up according to work status. The follow up was made at a minimum of 2 years after injury. Demographic data and scores from the occupational therapy assessments were compared for the two groups who were back to work or studies (BTW) and not back to work or studies (NBTW). RESULTS: Assessments of memory, visual perception and apraxia separated between the two groups BTW and NBTW. Logistic regression showed that memory score in combination with data on PADL made up the best predictive model. In a subgroup with 21 persons where data on IADL were added to memory and PADL the predictive value was even stronger. CONCLUSIONS: In this study we found that occupational therapy assessments were valuable in predicting failure to return to work or studies after brain injury. PMID- 11437200 TI - Rehabilitation an international perspective: a Zimbabwean experience. AB - PURPOSE/METHOD: The article discusses rehabilitation practices in Zimbabwe from both a local and international perspective. It considers the nature of formal and informal rehabilitation services in the country, and rehabilitation personnel training. RESULTS: Indigenous disability-related practices dominate the informal rehabilitation sector whereas modern rehabilitation practices characterize the formal rehabilitation sector. Rehabilitation clients make use of both the formal and informal rehabilitation systems. Persons with disabilities in Zimbabwean urban centres have more access to the formal rehabilitation service system than those in the rural areas. CONCLUSIONS: Rural communities may have more habilitation than rehabilitation, and are more accommodating of disability related differences. PMID- 11437201 TI - Wheelchair accessibility of public buildings in the central business district of Harare, Zimbabwe. AB - PURPOSE: To evaluate accessibility of wheelchairs into public buildings in the central business district of Harare, Zimbabwe and to identify architectural barriers faced by wheelchair users in public buildings. METHODS: This was a descriptive study of 20 public buildings in the Harare business district of Zimbabwe. Wheelchair accessibility was determined on a point scale based on specifications provided. Descriptive statistics of simple percentages and means were used to determine the level of compliance to the guidelines of the instrument, and wheelchair accessibility to the surveyed buildings. RESULTS: Of the various items surveyed, elevators recorded the highest average accessibility (83% compliance), while parking areas recorded the lowest average of 18%. The compliance score of ramps was 39%, while entrances were 71% compliant. Wheelchair accessibility to toilets was 51%. CONCLUSIONS: The poor compliance score for items such as parking areas, and ramps for wheelchair users is probably because the physically challenged persons and experts in the area of physical ability management are not consulted and involved in the design and construction of these buildings. High compliance of entrances and toilets may be purely coincidental and not for meeting the needs of wheelchair users. To improve on the wheelchair accessibility in public buildings, there should be liaison between the users the disabled, and various professionals (the occupational therapists, physiotherapists, politicians, engineers and architects). The findings of this study indicate a great challenge to the aforementioned professionals. Recommendations were made to increase wheelchair accessibility to public buildings. PMID- 11437202 TI - Paediatric rehabilitation. AB - More and more researchers are questioning the theoretical and scientific foundations as well as the efficacy and effects of many physiotherapy interventions. The same applies for many of the neurophysiological based interventions that are being used in paediatric rehabilitation. Opinions and views regarding the development of motor behaviour of infants and children are significantly changing. Paediatric interventionists should consider bringing their interventions and focus of treatment into agreement with changed scientific knowledge. Moreover, for almost all other medical problems in childhood, paediatric rehabilitation has little to offer but mostly miniaturized forms of adult treatment. It not only means that we have to make a paradigm shift, but also are in the need of a broader view on paediatric rehabilitation as a specialized professional activity. PMID- 11437203 TI - Contribution of soft tissue deformation below the sacrum to the measurement of total height loss in sitting. AB - This study investigated the contribution of soft tissue deformation below the sacrum (S) and vertical spinal creep to total height loss (THL) measured in sitting. Eight asymptomatic subjects (four males, four females) aged between 21 and 51 years were measured. Simultaneous measurement of THL and S were commenced after the subjects had been sitting for 5 min. THL was recorded while subjects were positioned in a seated stadiometer, which controlled their spinal posture. S was measured by placing an ultrasound transducer at the level of the top of the subject's sacrum. Over 25 min of sitting with loaded and unloaded interventions applied to their spine, different response characteristics between S and THL were noted. This study demonstrated that soft tissues below the sacrum could contribute up to 30% on average of total height loss. This suggests that researchers should take into account the soft tissue deformation outside the spine when studying vertical creep in sitting. PMID- 11437204 TI - Covariation between workplace physical and psychosocial stressors: evidence and implications for occupational health research and prevention. AB - There is increasing interest in distinguishing the effects of physical and psychosocial workplace stressors on the aetiology of work-related musculoskeletal disorders (MSD). Modest associations have been found between psychosocial stressors and MSD, such as intensive load, monotonous work and low job control. Interpretation of these results has been limited by likely covariation between physical and psychosocial stressors. This investigation examined exposure covariation among blue- and white-collar workers employed in a mass production manufacturing environment (N = 410). Physical stressors were assessed from questionnaire and accelerometry. Psychosocial stressors were assessed from questionnaire. Pearson and Spearman correlation coefficients were computed. An exploratory factor analysis procedure identified possible common factors linking specific physical and psychosocial stressors. Moderate to high correlations between some physical and psychosocial stressors showed evidence of covariation both across and within groups. Covariation was strongest among blue-collar production and low-status office workers. Factor analysis results showed considerable shared variance between some physical and psychosocial stressors, such as repetition and job control, suggesting that these disparate stressors manifest from common work organization factors that govern the structure of work. While recognizing the conceptual differences between physical and psychosocial stressors, these results call attention to the strong empirical relationships that can exist between some stressors in the workplace setting. To guard against ambiguous study findings that can occur when exposures are mixed, it is critical that future epidemiologic studies include information about the degree of association between task-level stressors. Future research on work organization determinants of task-level stressors, and their coincident occurrence in jobs with greater specialization, may provide promising new insights into the nature of risk for MSD and effective prevention strategies. PMID- 11437205 TI - Effects of sitting versus standing and scanner type on cashiers. AB - In the retail supermarket industry where cashiers perform repetitive, light manual material-handling tasks when scanning and handling products, reports of musculoskeletal disorders and discomfort are high. Ergonomics tradeoffs exist between sitting and standing postures, which are further confounded by the checkstand design and point-of-sale technology, such as the scanner. A laboratory experiment study was conducted to understand the effects of working position (sitting versus standing) and scanner type (bi-optic versus single window) on muscle activity, upper limb and spinal posture, and subjective preference of cashiers. Ten cashiers from a Dutch retailer participated in the study. Cashiers exhibited lower muscle activity in the neck and shoulders when standing and using a bi-optic scanner. Shoulder abduction was also less for standing conditions. In addition, all cashiers preferred using the bi-optic scanner with mixed preferences for sitting (n = 6) and standing (n = 4). Static loading of the muscles was relatively high compared with benchmarks, suggesting that during the task of scanning, cashiers may not have adequate recovery time to prevent fatigue. It is recommended that retailers integrate bi-optic scanners into standing checkstands to minimize postural stress, fatigue and discomfort in cashiers. PMID- 11437206 TI - Effects of dynamic office chairs on trunk kinematics, trunk extensor EMG and spinal shrinkage. AB - Seated work has been shown to constitute a risk factor for low-back pain. This is attributed to the prolonged and monotonous low-level mechanical load imposed by a seated posture. To evaluate the potential health effects with respect to the low back of office chairs with a movable seat and back rest, trunk kinematics, erector spinae EMG, spinal shrinkage and local discomfort were assessed in 10 subjects performing simulated office work. On three separate occasions subjects performed a 3 h task consisting of word processing, computer-aided design and reading. Three chairs were used, one with a fixed seat and back rest and two dynamic chairs, one with a seat and back rest movable in a fixed ratio with respect to each other, and one with a freely movable seat and back rest. Spinal shrinkage measurements showed a larger stature gain when working on the two dynamic chairs as compared with working on the chair with fixed seat and back rest. Trunk kinematics and erector spinae EMG were strongly affected by the task performed but not by the chair type. The results imply that dynamic office chairs offer a potential advantage over fixed chairs, but the effects of the task on the indicators of trunk load investigated were more pronounced than the effects of the chair. PMID- 11437207 TI - X-ray photoelectron spectroscopy (XPS) used to investigate the chemical interaction of synthesized polyalkenoic acid with enamel and synthetic hydroxyapatite. PMID- 11437208 TI - Fluoride release from glass ionomer activated with fluoride solutions. AB - The mechanism of the fluoride release from glass-ionomer cements (GICs) is not yet completely understood, due to the complexity of these systems. The objective of the present study was to investigate the fluoride and alkali metal ion release from a relatively simple GIC formulation with fluoride- and alkali metal-free glass and activated with a NaF or KF solution. The set formulations were eluted during 168 days in water at 37 degrees C. The cumulative fluoride release was the result of an initial high release that ceased after some time and a prolonged but slow release, both of which increased with increasing fluoride concentrations in the activating solution, independently of the type of alkali metal. Maturation prior to elution decreased the fluoride and slowed the alkali metal release. The release of these ions was not (completely) correlated. The results suggest that the release process is due not only to a loss of relatively loosely bound fluoride in the cement matrix, but also to the release of fluoride which becomes strongly bound during the setting reaction and induces a long-term release. PMID- 11437209 TI - Remineralization of caries lesions extending into dentin. AB - Remineralization is one aspect of the overall process of tooth decay. However, it is primarily studied in shallow lesions. The aim of this study was to explore whether caries lesions in enamel and extending into the dentin can be remineralized. A single-section model was developed for the longitudinal and non destructive monitoring of changes in enamel and dentin. Lesions at least 200 microm into dentin were formed in undersaturated acetate buffers. Next, the lesions were divided into groups (three treatment and one control) and remineralized. The treatments were: weekly immersion in 1,000 ppm fluoride, single treatment with methanehydroxybisphosphonate, and a constant level of 1 ppm fluoride. De- and remineralization was assessed by transverse microradiography. Remineralization was observed in enamel, but also in dentin, indicating that, deep into dentin, the pores become supersaturated to apatite formation. Treatments affected remineralization only in the outer part of enamel. Both findings are explained by a relatively fast diffusion of mineral ions, with precipitation being rate-limiting. The results suggest that dentin remineralization, underneath enamel, can be achieved and could possibly be used in clinical treatment strategies. PMID- 11437210 TI - Distribution and excretion of TEGDMA in guinea pigs and mice. AB - The monomer triethyleneglycoldimethacrylate (TEGDMA) is used as a diluent in many resin-based dental materials. It was previously shown in vitro that TEGDMA was released into the adjacent biophase from such materials during the first days after placement. In this study, the uptake, distribution, and excretion of 14C TEGDMA applied via gastric, intradermal, and intravenous administration at dose levels well above those encountered in dental care were examined in vivo in guinea pigs and mice as a test of the hypothesis that TEGDMA reaches cytotoxic levels in mammalian tissues. 14C-TEGDMA was taken up rapidly from the stomach and small intestine after gastric administration in both species and was widely distributed in the body following administration by each route. Most 14C was excreted within one day as 14CO2. The peak equivalent TEGDMA levels in all mouse and guinea pig tissues examined were at least 1000-fold less than known toxic levels. The study therefore did not support the hypothesis. PMID- 11437211 TI - Low magnitude of tensile strain inhibits IL-1beta-dependent induction of pro inflammatory cytokines and induces synthesis of IL-10 in human periodontal ligament cells in vitro. AB - Applied mechanical loading induces inflammation in the periodontal ligament (PDL). However, the mechanisms involved in bone deposition at tension sites in an inflammatory environment are not clear. Here, in an in vitro model system, we show that equibiaxial tensile strain of low magnitude (TENS) provokes potent anti inflammatory signals in PDL cells. TENS inhibits IL-1beta-induced synthesis of IL 1beta, IL-6, and IL-8 by inhibiting their mRNA expression, and thus significantly suppresses the amplification of IL-1beta-induced inflammatory responses in PDL cells. Additionally, as an anti-inflammatory signal, TENS induces IL-10 synthesis in the presence and absence of IL-1beta. These observations are the first to demonstrate that TENS antagonizes IL-1beta actions on PDL cells by (i) inhibiting IL-1beta-induced transcriptional regulation of proinflammatory cytokines, and (ii) inducing synthesis of IL-10, which may post-transcriptionally suppress the synthesis of pro-inflammatory cytokines. PMID- 11437212 TI - Impairment of gingival fibroblast adherence by IL-6/sIL-6R. AB - Interleukin-6 (IL-6) binds to human gingival fibroblasts (HGF) in the presence of a soluble form of IL-6 receptor (sIL-6R). We investigated the effects of IL-6 on the functions of HGF in the presence of sIL-6R. HGF changed their morphology from spindle-shaped to round, and detached from the culture dish by stimulation with IL-6/sIL-6R. In this condition, a signal transducer gp130 and a transcription factor Stat3 were phosphorylated, resulting in activation of transcription factors Stat3 and C/EBPbeta. Cytoskeletal beta-actin and adhesion molecule integrin-alpha5, a subunit of alpha5beta1 integrin (VLA-5), were found to possess potential binding domains for these transcription factors in their promoters. Accumulation of beta-actin and integrin-alpha5 mRNA decreased, contrary to the expectation of the induction of gene transcription. Furthermore, the decrease in their mRNAs was associated with reduced expression of both actin and VLA-5 proteins. These results suggest that the expression of VLA-5 and actin was down regulated in HGF through an IL-6 signaling pathway, resulting in impairment of HGF adherence. PMID- 11437213 TI - Preferential utilization of dipeptides by Porphyromonas gingivalis. AB - Although Porphyromonas gingivalis is known to utilize peptides preferentially, instead of free amino acids, as the source of energy and cell material, there is only limited information on what sizes and kinds of peptide this bacterium preferentially utilizes. In this study, therefore, we tested aspartate or glutamate monopolymers consisting of from 2 to 100 amino acids as metabolic substrates for P. gingivalis. The washed cells of P. gingivalis consumed aspartylaspartate and glutamylglutamate, and produced large amounts of ammonia and organic acids such as propionate and butyrate, while the cells formed only small amounts of end-products from aspartate, glutamate, and other peptides longer than a dipeptide. P. gingivalis also metabolized valylvaline and leucylleucine and produced isobutyrate and isovalerate, respectively, only in the presence of aspartylaspartate or glutamylglutamate. This suggests a metabolic linkage between these dipeptides. These results clearly indicate that P. gingivalis utilizes dipeptides preferentially as its metabolic substrates. PMID- 11437214 TI - Condylar motion in patients with reduced anterior disc displacement. AB - The influence of reduced anterior disc displacement on condylar motion has not been fully examined in young adults. Reduced anterior disc displacement was hypothesized to inhibit condylar motion. Using a six-degrees-of-freedom jaw tracking system, we recorded bilateral condylar motion during maximum open-close jaw movement and gum-chewing on both sides in ten young adults with unilateral reduced anterior disc displacement and in ten control subjects without temporomandibular disorders. The bilateral condylar motion during both maximum open-close jaw movement and chewing on the disc-displacement side was inhibited in the test group. The condylar motion on the disc-displacement side during chewing on the non-disc-displacement side was also inhibited in the test group. These results suggest that the limitation of condylar motion on the disc displacement side may influence condylar motion on the non-disc-displacement side during maximum open-close jaw movement, and mastication on the disc-displacement side in young adults. PMID- 11437215 TI - Conofocal microscopy study of undisturbed and chlorhexidine-treated dental biofilm. AB - Culturing of dispersed plaque samples and vitality staining of plaque smears are the most commonly used methods for evaluating the effects of antimicrobials on dental plaque. The visualization of the antimicrobial action on oral biofilm present on the substrate surface (in situ) would add valuable information to the existing knowledge about the treatment effects. This study aimed at combining the advantage of confocal laser scanning microscopy (CLSM) to visualize plaque non destructively with a vitality staining technique to assess the immediate bactericidal effect of chlorhexidine (CHX) on biofilm. Three 200-microm-wide grooves were cut into bovine dentin discs for plaque accumulation. The discs were worn by six subjects for 6, 24, and 48 hrs, then broken into halves, one of which received a one-minute extraoral 0.2% CHX treatment, while the other served as control. Both halves were stained for vital fluorescence measurements and visualized by CLSM. Plaque vitality (in %) was quantified by image analysis in three plaque layers-outer, middle, and inner. The CHX effect was significant in six-hour samples (p < 0.001) and only in the outer layer of the 48-hour plaque (p < 0.001), demonstrating a resistant nature of dental biofilm to a single CHX treatment. With the present approach, we have shown that it is possible to visualize and quantitate the antimicrobial treatment effect on biofilm still present on the substrate on which it was grown. PMID- 11437216 TI - Spontaneous fluctuations in the concentrations of oral sulfur-containing gases. AB - Breath hydrogen sulfide (H2S) and methyl-mercaptan (CH3SH) concentrations are used as quantitative indicators of halitosis. However, measurements of these gases in duplicate oral samplings often show poor reproducibility. To determine if this poor reproducibility is an artifact of the collection/analytical procedure or a true biological phenomenon, we used a standardized technique to collect from 20 to 30 oral gas samples at two-minute intervals from 11 healthy subjects. The samples were analyzed for sulfur gases and CO2. Sizable variations in H2S and CH3SH concentrations were not associated with alterations in CO2, indicating that the variations did not reflect variable contamination with atmospheric or pulmonary gas. In addition, fluctuations in H2S and CH3SH were not identical and often were not random. We conclude that minute-to-minute variability in oral sulfur gas concentrations is a true biological phenomenon. This fluctuation complicates experimental studies designed to show that interventions alter halitosis. PMID- 11437217 TI - Characteristics of incisor-premolar hypodontia in families. AB - Nonsyndromic tooth agenesis is a genetically and phenotypically heterogenous condition. It is generally assumed that different phenotypic forms are caused by different mutated genes. We analyzed inheritance and phenotype of hypodontia and dental anomalies in 214 family members in three generations of 11 probands collected for genetic linkage study on incisor-premolar hypodontia (IPH). Our analysis confirms the autosomal-dominant transmission with reduced penetrance of IPH. The prevalence of hypodontia and/or peg-shaped teeth was over 40% in first- and second-degree relatives and 18% in first cousins of the probands. Four of nine noted obligate carriers of hypodontia gene had dental anomalies, including small upper lateral incisors, ectopic canines, taurodontism, and rotated premolars. These anomalies were also observed at higher than normal frequency in relatives affected with hypodontia. We conclude that incisor-premolar hypodontia is a genetic condition with autosomal-dominant transmission and that it is associated with several other dental abnormalities. PMID- 11437218 TI - An Oral Health Survey in Southern China, 1997: preface. PMID- 11437219 TI - An Oral Health Survey in Southern China, 1997: background and methodology. AB - Relatively limited information is available about the oral health in the Guangdong Province of Southern China, the closest neighbor to Hong Kong. The study intended to explore the oral health status, knowledge, attitudes, and dental care utilization in the Guangdong population as a basis for formulating strategies for oral health prevention and treatment. Through multi-stage stratified and quota-sampling, individuals from urban as well as rural communities were selected (5- to 6-; 12-; 35- to 44-; and 65- to 74-year-olds; total N = 6251). Structured interviews and clinical examinations were performed. Inter-examiner reliability was high (kappa = 0.60-0.96). Samples of community water were taken for fluoride assessment. Overall, the sample surveyed was acceptably representative of the population, with some under-representation of rural residents and agricultural workers. Re-weighting was performed in appropriate analyses. The data analysis model used in the Second International Collaborative Study was used as a guide for the present data analysis. This approach has not previously been used on a Mainland Chinese population. PMID- 11437220 TI - Oral health status and oral health behaviors in Chinese Children. AB - The objectives of the study were to describe the oral health status and treatment needs of the 5- to 6-year-old and 12-year-old children in Southern China; to describe the patterns of oral health behaviors, knowledge, and attitudes among the 12-year-olds; and to assess the effects of socio-behavioral factors on the 12 year-old children's dental caries experiences. The study sample was comprised of 1,587 5- to 6-year-old and 1,576 12-year-old urban and rural schoolchildren living in Guangdong Province. Three calibrated dentists clinically examined the children, and trained interviewers interviewed the 12-year-olds. Caries prevalence of the 5- to 6-year-old children was high (urban 78% vs. rural 86%); the mean dmft of the urban and rural children was 4.8 and 7.0, respectively. The caries prevalence and mean DMFT score of the 12-year-olds were 41% and 0.9 (urban) and 42% and 0.9 (rural). Only 2% of the 12-year-olds exhibited no calculus or gingival bleeding, while more than 70% had calculus. In conclusion, there is an urgent need for establishing caries-preventive activities for preschool children. The prevalence of caries among the 12-year-olds was not high, but their periodontal condition was unsatisfactory. Knowledge about gum bleeding and the use of fluoride was low. More oral health education activities should be organized, especially for the rural children. PMID- 11437221 TI - Oral health knowledge, attitudes, and practices of Chinese adults. AB - Only recently have behavioral aspects been taken into consideration in oral health studies in China; thus, relevant information on perceptions in adult Chinese about oral health care is inadequate. In 1996-97, a large-scale oral epidemiological study was conducted in Southern China. Among its objectives were to describe the oral health knowledge, attitudes, and practices of the urban and rural adults living in Southern China and to analyze the influence of selected demographic and socio-economic factors. From 8 urban and 8 rural communities in Guangdong Province, 1,573 35- to 44-year-olds and 1,515 65- to 74-year-olds were sampled. Specially trained interviewers from the staff of local government or hospitals conducted face-to-face structured interviews. Almost all of the middle aged and more than 90% of the dentate elderly surveyed claimed that they brushed their teeth every day and used toothpaste during toothbrushing, but awareness about fluoride content was lacking. The respondents had poor oral health knowledge but positive attitudes toward oral health, providing a basis for more community-based oral health education programs, especially targeting adults who are less-well-educated and have fewer socio-economic advantages. PMID- 11437222 TI - Utilization of dental services in Southern China. AB - A population's utilization of dental services is an important parameter in oral health care planning, which has rarely been studied in China. The objectives of this report were to describe the dental service utilization pattern of middle aged and elderly Chinese and to analyze the influence of selected variables on the use of dental services. A Guangdong Province population of 1,573 35- to 44 year-olds and 1,515 65- to 74-year-olds recruited from urban and rural communities was interviewed in their local dialect. It was found that 23% of the middle-aged and 24% of the elderly subjects had visited a dentist within the preceding year. The two most commonly cited reasons for not having seen a dentist for at least 3 years were: no perceived need, and no serious dental problems. Among subjects who had visited a dentist within 3 years, the 3 most commonly received treatments were: fillings, extractions, and dental prostheses. Furthermore, a logistic regression analysis showed that women, subjects who lived in urban areas, were better educated, were wealthier, and had better oral health knowledge were more likely to be a recent dental service user. In conclusion, dental service utilization among the adult Southern Chinese was found to be low, problem-driven, and influenced by some socio-economic factors. PMID- 11437223 TI - Coronal and root caries in Southern Chinese adults. AB - Southern China is the most prosperous part of China, but information useful for oral care planning is very limited. A large-scale epidemiological survey was conducted in 1996-97. The objectives of this report were to describe the coronal and root caries of the adult Southern Chinese and to analyze the influence of selected demographic and socio-economic factors on the disease pattern. A total sample of 1,573 35- to 44-year-olds and 1,515 65- to 74-year-olds from 8 urban and 8 rural survey sites in Guangdong Province participated in an oral health interview and underwent clinical examination. World Health Organization examination procedures and diagnostic criteria were used. The weighted mean DMFT scores of the middle-aged and the elderly subjects were 4.8 and 16.1, respectively. People living in rural areas had a higher DMFT score than those living in urban areas (4.9 vs. 4.3 in the 35- to 44-year-olds and 16.5 vs. 14.7 in the 65- to 74-year-olds). In both age groups, MT was the major component of the DMFT score. Analysis of covariance showed that women and those who were economically less well off had higher DMFT scores in both age groups. The weighted prevalence rates of decayed/filled roots were 12% and 37%, with a mean of 0.2 and 0.7 teeth affected, in the middle-aged and the elderly, respectively. In conclusion, socio-economic factors had a considerable effect on the dental caries status of adults in Southern China. PMID- 11437224 TI - Periodontal conditions in adult Southern Chinese. AB - Periodontal diseases remain a concern in adult populations, but there have been no systematic descriptions of periodontal conditions, which have included periodontal attachment loss, among Southern Chinese. The main aim of this report is to describe the periodontal conditions in adult Chinese and to investigate factors that may have influenced periodontal attachment loss. Subjects were recruited from urban and rural survey sites throughout Guangdong. In total, 1,572 35- to 44-year-old subjects and 1,286 65- to 74-year-old subjects completed a structured interview and underwent a periodontal examination, which included the Community Periodontal Index (CPI) and periodontal Attachment Loss (ALoss) recordings. Calculus was found as the highest CPI score in 61-68% of the 35- to 44-year-old subjects and in 54-57% of the 65- to 74-year-olds. Shallow pockets were found as the highest CPI score in about one-third of both the urban and the rural subjects in both age groups, and deep pockets in 3-7% of the subjects. ALoss was more prevalent than pockets in both age groups. On the basis of the ALoss recordings, about one-third of the subjects in both age groups were categorized as exhibiting considerable ALoss for their age. In both age groups, logistic regression analysis indicated that being male, wearing partial dentures, and reporting less frequent toothbrushing were found to be associated with considerable ALoss. PMID- 11437225 TI - Oral mucosal lesions in adult Chinese. AB - The objectives of this analysis were to determine the prevalence of oral mucosal lesions (OML) among adults in Southern China and to determine possible associations between OML and reported tobacco-smoking and alcohol-drinking habits. The sample consisted of 1,573 35- to 44-year-old and 1,515 65- to 74-year old Chinese from both urban and rural areas of Guangdong Province. The subjects were interviewed by trained interviewers and underwent a clinical examination of the oral mucosa performed according to WHO guidelines. A specially prepared color atlas of OML was used for lesion recognition and confirmation during the survey. Among the 35- to 44-year-old subjects, the overall prevalence of OML was found to be 13% in urban men, 6% in urban women, 15% in rural men, and 4% in rural women. The corresponding figures for the 65- to 74-year-olds were 22%, 12%, 26%, and 19%. Tongue lesions and white lesions were relatively common, but denture-related lesions were not. No malignancies or erythroplakia was observed. Age and gender were found to relate to the occurrence of OML. In elderly men, smoking was significantly (p < 0.05) associated with some white lesions and tongue lesions, and alcohol drinking was also associated with some white lesions. In conclusion, tongue lesions and white lesions were relatively common in men, but pre-cancerous lesions were not prevalent. PMID- 11437226 TI - Tooth loss, occluding pairs, and prosthetic status of Chinese adults. AB - The objectives of this analysis were to describe the numbers of teeth lost and those indicated for extraction, dental prosthetic status, and occluding tooth pairs among adults in Southern China and to investigate factors affecting tooth mortality. A sample consisting of 1,573 35- to 44-year-old and 1,515 65- to 74 year-old urban and rural Chinese was examined clinically by calibrated examiners. Teeth not present for any reason were defined as missing (MT). The sum of MT and teeth indicated for extraction (IMT) was defined as adjusted missing teeth (AMT). Only 4.4% of the urban and 3.4% of the rural 65- to 74-year-olds were edentulous. MT were not significantly different between urban and rural residents, but more IMT were found in rural residents. Women, those who had a lower education level, and those who were less wealthy had more AMT in both age groups. Smoking was found to be associated with AMT in the 35- to 44-year-olds. Eighteen percent of the urban and 27% of the rural elderly had no occluding tooth pairs after IMT were accounted for, and only 38% and 30%, respectively, had 10 or more occluding pairs. Fifty-one percent of the urban and 43% of the rural elderly were found to have some form of dental prostheses in either jaw. A partial denture was more common in urban residents in both age groups. PMID- 11437227 TI - Introduction: assembly, remodeling and modification of chromatin. PMID- 11437228 TI - Chromatin assembly during S phase: contributions from histone deposition, DNA replication and the cell division cycle. AB - During S phase of the eukaryotic cell division cycle, newly replicated DNA is rapidly assembled into chromatin. Newly synthesised histones form complexes with chromatin assembly factors, mediating their deposition onto nascent DNA and their assembly into nucleosomes. Chromatin assembly factor 1, CAF-1, is a specialised assembly factor that targets these histones to replicating DNA by association with the replication fork associated protein, proliferating cell nuclear antigen, PCNA. Nucleosomes are further organised into ordered arrays along the DNA by the activity of ATP-dependent chromatin assembly and spacing factors such as ATP utilising chromatin assembly and remodelling factor ACE An additional level of controlling chromatin assembly pathways has become apparent by the observation of functional requirements for cyclin-dependent protein kinases, casein kinase II and protein phosphatases. In this review, we will discuss replication-associated histone deposition and nucleosome assembly pathways, and we will focus in particular on how nucleosome assembly is linked to DNA replication and how it may be regulated by the cell cycle control machinery. PMID- 11437229 TI - ATP-dependent chromatin remodeling activities. AB - Genetic and biochemical approaches have indicated that the packaging of DNA into chromatin can be repressive to transcription. ATP-dependent chromatin remodelling activities can facilitate transcription from chromatin templates. Consistent with this, biochemical assays have shown that the action of ATP-dependent chromatin remodelling activities increase the accessibility of DNA within chromatin templates. However more recent functional studies suggest that many ATP-dependent chromatin remodelling activities can also function as repressors of transcription. Here we review recent advances to our understanding of the biological function of these complexes. We then consider some of the mechanisms by which ATP-dependent chromatin remodelling activities together with other forms of chromatin remodelling or modifying enzymes may act to regulate genomic accessibility either positively or negatively. PMID- 11437230 TI - Mammalian histone acetyltransferases and their complexes. AB - One of the key questions in the current molecular genetics of eukaryotes is how genetic information is retrieved from tightly packed chromatin. Acetylation of core histone N-termini is implicated in the regulation of chromatin function, and I summarize what is known about the mammalian enzymes that promote this posttranslational histone modification. Chromatin is important in gene expression not only because of the accessibility problem that it poses for the transcriptional machinery but also with regard to the phenomenon of chromatin memory, i.e. the ability of alternative chromatin states to be maintained through many cell divisions. This phenomenon is believed to be central to epigenetic inheritance, an important concept in developmental biology, which is also emerging as a contributing factor in cancer and other health disorders. Analyses of the composition of large multiprotein acetyltransferase complexes suggest their role in the mechanisms of epigenetic inheritance. The review will discuss some models pertinent to this function of histone acetyltransferases. PMID- 11437231 TI - Structure and function of histone acetyltransferases. AB - Histone acetyltranferase (HAT) enzymes are the catalytic subunit of large multisubunit HAT complexes that acetylate the epsilon-amino group of specific lysine residues on histone tails to promote transcriptional activation. Recent structural and functional studies on the divergent HAT enzymes Gcn5/PCAF, Esa1 and Hat1 have provided new insights into the underlying mechanism of histone binding and acetylation by HAT proteins. The three HAT enzymes contain a structurally conserved core domain that plays a functionally conserved role in binding the coenzyme A cofactor and in harboring the putative general base for catalysis. Structurally variable N- and C-terminal domains appear to contain a related scaffold that mediates histone substrate binding. These data provide a framework for understanding the structure and function of other more divergent HAT proteins such as TAF(II)250 and CBP/p300, and provides a starting point for understanding how HAT proteins may cooperate with other factors within in vivo HAT complexes to promote transcriptional activation. PMID- 11437232 TI - Histone acetylation: plants and fungi as model systems for the investigation of histone deacetylases. AB - The basic element of chromatin is the nucleosome. Histones H4, H3, H2A and H2B form the core histone octamer by protein-protein interactions of their folded domains. The free, flexible N-terminal extensions of the histones protrude from the nuclesome; they contain conserved lysines undergoing posttranslational acetylation. Histone acetyltransferases (HATs) transfer the acetyl moiety of acetyl-coenzyme A to the epsilon-amino group; this reaction is reverted by histone deacetylases (HDACs). The dynamic equilibrium of the acetylation/deacetylation reaction varies throughout the genome; some regions in chromatin undergo rapid acetylation/deacetylation, whereas others are fixed in a certain acetylation state without significant changes. In general, chromatin regions engaged in transcription display dynamic acetylation, i.e. HATs and HDACs are recruited to these regions. Higher plants and fungi have considerably contributed to the unraveling of the multiplicity of HDACs; in particular, plants possess HDACs that have so far not been identified in animal cells. PMID- 11437233 TI - Emerging connections between DNA methylation and histone acetylation. AB - Modifications of both DNA and chromatin can affect gene expression and lead to gene silencing. Evidence of links between DNA methylation and histone hypoacetylation is accumulating. Several proteins that specifically bind to methylated DNA are associated with complexes that include histone deacetylases (HDACs). In addition, DNA methyltransferases of mammals appear to interact with HDACs. Experiments with animal cells have shown that HDACs are responsible for part of the repressive effect of DNA methylation. Evidence was found in Neurospora that protein acetylation can in some cases affect DNA methylation. The available data suggest that the roles of DNA methylation and histone hypoacetylation, and their relationship with each other, can vary, even within an organism. Some open questions in this emerging field that should be answered in the near future are discussed. PMID- 11437234 TI - Histone acetylation and disease. AB - Differential acetylation of histones and transcription factors plays an important regulatory role in developmental processes, proliferation and differentiation. Aberrant acetylation or deacetylation leads to such diverse disorders as leukemia, epithelial cancers, fragile X syndrome and Rubinstein-Taybi syndrome. The various groups of histone acetyltransferases (CBP/p300, GNAT, MYST, nuclear receptor coactivators and TAFII250) and histone deacetylases are surveyed with regard to their possible or known involvement in cancer progression and human developmental disorders. Current treatment strategies are discussed, which are still mostly limited to histone deacetylase inhibitors such as trichostatin A and butyrate. PMID- 11437235 TI - Cytochromes P450 and metabolism of xenobiotics. AB - Cytochromes P450 (henceforth P450s) are involved in a variety of metabolic and biosynthetic processes. The number of known P450 enzymes exceeds 1000, while the endogenous substrates of most of them remain unknown. All P450 enzymes exhibit similarity in their structure and general mechanism of action; however, there are significant differences in the detailed function of individual enzymes as well as in the structures and properties of their active sites. This review discusses the properties of the most important P450 enzymes taking part in drug metabolism in humans. P450 3A4 is of paramount importance, because it is the most abundant P450 in the human liver and is known to metabolize the majority of drugs whose biotransformation is known. Genetically dependent variabilities of individual P450 activities and levels are described, documenting the importance of pharmacogenetics aimed at explaining differences in the response of the organism to various drugs. PMID- 11437236 TI - Nerve growth factor: structure and function. AB - Neurotrophins are critical for the development and maintenance of the peripheral and central nervous system. These highly homologous, homodimeric growth factors control cell survival, differentiation, growth cessation, and apoptosis of sensory neurons. The biological functions of the neurotrophins are mediated through two classes of cell surface receptors, the Trk receptors and the p75 neurotrophin receptor (p75NTR). Nerve growth factor (NGF), the best characterized member of the neurotrophin family, sends its survival signals through activation of TrkA and can induce cell death by binding to p75NTR. Recent domain deletion and mutagenesis studies have identified the membrane-proximal domain of the Trks as necessary and sufficient for ligand binding. Crystal structures of this domain of TrkA, TrkB, and TrkC, and an alanine scanning analysis of this domain of TrkA and TrkC have allowed identification of the ligand-binding site. The recent crystal structure of the complex between NGF and the ligand-binding domain of TrkA defines the orientation of NGF in the signaling complex, and eludicates the structural basis for binding and specificity in the family. Further structural work on NGF-TrkA-p7SNTR complexes will be necessary to address the many remaining questions in this complex signaling system. PMID- 11437237 TI - The glycinergic inhibitory synapse. AB - Glycine is one of the most important inhibitory neurotransmitters in the spinal cord and the brainstem, and glycinergic synapses have a well-established role in the regulation of locomotor behavior. Research over the last 15 years has yielded new insights on glycine neurotransmission. Glycinergic synapses are now known not to be restricted to the spinal cord and the brainstem. Presynaptic machinery for glycine release and uptake, the structure and function of postsynaptic receptors and the factors (both pre- and postsynaptic) which control the strength of glycinergic inhibition have been extensively studied. It is now established that glycinergic synapses can be excitatory in the immature brain and that some inhibitory synapses can corelease gamma-aminobutyric acid (GABA) and glycine. Moreover, the presence of glycine transporters on glial cells and the capacity of these cells to release glycine suggest that glycine may also act as a neuromodulator. Extensive molecular studies have revealed the presence of distinct subtypes of postsynaptic glycine receptors with different functional properties. Mechanisms of glycine receptors aggregation at postsynaptic sites during development are better understood and functional implications of variation in receptor number between postsynaptic sites are partly elucidated. Mutations of glycine receptor subunits have been shown to underly some human locomotor disorders, including the startle disease. Clearly, recent work on glycine receptor channels and the synapses at which they mediate inhibitory signalling in both young and adult animals necessitates an update of our vision of glycinergic inhibitory transmission. PMID- 11437238 TI - Analysing structure-function relationships with biosensors. AB - Elucidating the nature of the relationship between the structure and function of biomolecules remains one of the major challenges in biology. Biomolecules are dynamic entities that possess a variety of structures, and their functions at the molecular, cellular and organismic levels are quite different. Since there is no single causal link between structure and function, the search should be for correlations rather than causal relations. Biosensor instruments based on surface plasmon resonance are widely used for establishing correlations between the chemical structure of binding sites and their binding activity. Mutagenesis studies have shown that only a small percentage of the residues located in a binding site contribute to the binding energy. Since substitutions in residues located far away from the binding site are able to affect binding activity, this greatly complicates the rational design of proteins endowed with improved functions. However, biosensors can be used to determine and predict the influence of the chemical environment and of the structure of a ligand on binding kinetics. PMID- 11437239 TI - On the existence of cytokines in invertebrates. AB - Based on the assumption that invertebrates, like vertebrates, possess factors regulating responses to infection or wounding, studies dealing with the evolution of immunity have focussed on the isolation and characterisation of putative cytokine-related molecules from invertebrates. Until recently, most of our knowledge of cytokine- and cytokine receptor-like molecules in invertebrates relies on functional assays and similarities at the physicochemical level. As such, a phylogenetic relationship between invertebrate cytokine-like molecules and vertebrate counterparts could not be convincingly demonstrated. Recent genomic sequence analyses of interleukin-1-receptor-related molecules, that is Toll-like receptors, and members of the transforming growth factor-beta superfamily suggest that the innate immune system of invertebrates and vertebrates evolved independently. In addition, data from protochordates and annelids suggest that invertebrate cytokine-like molecules and vertebrate factors do not have the same evolutionary origin. We propose instead that the convergence of function of invertebrate cytokine-like molecules and vertebrate counterparts involved in innate immune defences may be based on similar lectin-like activities. PMID- 11437240 TI - Allometry of mammalian cellular oxygen consumption. AB - In the 1930s, Max Kleiber and Samuel Brody established that the interspecies correlation between mammalian body mass and metabolic rate (alphaM(0.75)) cannot be explained (solely) by whole body surface area (alphaM(0.66)) to volume ratios. Metabolic considerations must also be taken into account. Decreases in the proportion of visceral organ mass to whole body mass can account for some of the whole body metabolic differences. However, superimposed upon these anatomical differences, the metabolism of tissues and cells has been demonstrated to decrease with increasing body mass. These decreases in oxygen consumption rates (with increasing body mass) in cells and tissues can be explained by a decrease in ATP turnover and mitochondrial density and an increase in mitochondrial functional efficiency (decrease in proton leak). The majority of the proton leak differences reflect differences in mitochondrial inner membrane surface area. Indeed, liver metabolism correlates directly with liver mitochondrial inner membrane surface area. Apart from being a significant contributor (approximately 25%) to basal metabolism, mitochondrial proton leak is a major factor determining the differences in basal metabolism between mammals of different body mass. PMID- 11437241 TI - Purification, cloning and characterisation of odorant- and pheromone-binding proteins from pig nasal epithelium. AB - Two distinct classes of lipocalin isoforms (OBP-IIs and OBP-IIIs) were purified and identified from porcine nasal mucosa of male and female individuals. Using primers designed on their N-terminal sequence, the complete primary structures of the mature polypeptides were determined. Mass spectrometry analysis confirmed the identity of the cDNA-derived sequences and provided information regarding their post-translational modifications. These species strongly resemble a lipocalin expressed by von Ebner's gland and salivary lipocalins carrying sex-specific pheromones secreted only by the boar's submaxillary glands. Both OBP-IIs and OBP IIIs present two cysteines paired in a disulphide bond; the remaining residues occur in a reduced form. In addition, OBP-IIIs are heavily glycosylated and markedly different in their glycan moiety from the salivary lipocalins. A three dimensional model is proposed based on protein species with known structure. Like salivary lipocalins, OBP-IIIs bind a number of odorant molecules, with highest affinity for the specific pheromone 5alpha-androst-16-en-3-one. The high similarity between OBPs from the nasal area and lipocalins from secretory glands suggests a common function in binding the same pheromonal ligands, the latter carrying chemical messages into the environment the former delivering them to specific receptors. PMID- 11437242 TI - Probing the penicillin sidechain selectivity of recombinant deacetoxycephalosporin C synthase. AB - Deacetoxycephalosporin C synthase from Streptomyces clavuligerus catalyses the conversion of the five-membered penicillin ring to the unsaturated six-membered cephem ring of deacetoxycephalosporin C. The effects on enzyme activity of the penicillin substrate sidechain and various cofactors were investigated using a continuous spectrophotometric assay. The conversion of penicillin G to phenylacetyl-7-aminodeacetoxycephalo sporanic acid (G-7-ADCA) was confirmed, and further details of the reaction were elucidated. The conversion of ampicillin to cephalexin was faster than that of acetyl-6-APA to acetyl-7-ADCA kcat = 0.120 +/- 0.001 s(-1) versus 0.035 +/- 0.001 s(-1), but they had similar Km values: 4.86 +/ 0.12 and 3.28 +/- 0.26 mM, respectively. Amoxycillin and penicillin V were also converted at low levels. Conversion was not detected for penicillanate, 6 aminopenicillanate, carbenicillin, temocillin, ticarcillin or benzylpenicilloic acid, suggesting that the enzyme has a relatively strict selectivity for the sidechain of the penicillin substrate. PMID- 11437243 TI - Neurodegeneration changes in primary central nervous system neurons transfected with the Alzheimer-associated neuronal thread protein gene. AB - The AD7c-NTP gene is over-expressed in brains with Alzheimer's disease (AD), and increased levels of the corresponding protein are detectable in cortical neurons, brain tissue extracts, cerebrospinal fluid, and urine beginning early in the course of AD neurodegeneration. In the present study, we utilized a novel method to transfect post-mitotic primary neuronal cell cultures, and demonstrated that over-expression of the AD7c-NTP gene causes cell death and neuritic sprouting, two prominent abnormalities associated with AD. These results provide further evidence that aberrantly increased AD7c-NTP expression may have a role in AD-type neurodegeneration. In addition, we demonstrate that primary post-mitotic neurons can be efficiently transfected with conventional recombinant plasmid DNA to evaluate the effects of gene over-expression in relevant in vitro models. PMID- 11437244 TI - Histamine deficiency suppresses murine haptoglobin production and modifies hepatic protein tyrosine phosphorylation. AB - Histidine decarboxylase (HDC) synthesizes endogenous histamine from histidine in mammals. HDC-deficient mice (HDC-/-), if kept on a histamine-free diet, have no histamine in their tissues. HDC-/- mice show multiple phenotypes. In this study we show that both the constitutively expressed and turpentine-induced level of an acute-phase protein, haptoglobin, is significantly lower in the serum of HDC-/- mice compared to that of wild-type animals. This effect was abolished if HDC gene targeted mice received histamine-rich food. No differences were found when lipopolysaccharide (LPS) was used to induce the acute-phase reaction. Using specific antibodies to phosphorylated tyrosine, we showed that protein tyrosine phosphorylation (Y-P) of approximately 50- and 26- to 27-kDa liver proteins is significantly decreased in HDC-/- mice, but that the difference was largely diminished if the animals were kept on a histamine-rich diet, suggesting that the phenotype with lower haptoglobin production is diet inducible. Upon in vivo treatment with LPS, Y-P band intensity decreased, regardless of the presence or absence of histamine. Identification of elements of the signalling pathway with decreased phosphorylation may elucidate the molecular background of the effect of endogenous histamine in the hepatic acute-phase reaction. PMID- 11437245 TI - Molecular and biochemical characterization of three aromatic polyketide synthase genes from Rubus idaeus. AB - Three polyketide synthase genes (PKS1, PKS2, PKS3) from cell suspension cultures of raspberry (Rubus idaeus L. cv. Royalty) were characterized. They showed high similarity in both their nucleotide and deduced amino acid sequences. All three proteins contain the amino acid residues identified in previous work as essential for chalcone synthase (CHS) function. Enzyme activities were investigated after heterologous expression in Escherichia coli. RiPKS1 is a typical naringenin CHS that synthesizes the chalcone as the main reaction product, and p coumaryltriacetic acid lactone (CTAL) as a minor by-product. RiPKS3 differed from RiPKS1 in four positions (K49R, M64R, P120L, V188A), and the products in vitro were predominantly CTAL and low levels of chalcone. RiPKS2 had the same four differences from RiPKS1 as RiPKS3, but in addition two further exchanges (R259H, F344L), and the protein had no detectable enzyme activity. Experiments with RiPKS1 containing either 259H or 344L showed that each of the exchanges was sufficient to completely eliminate enzyme activity. These experiments identify amino acid residues in CHS which are important for folding of the tetraketide intermediate to the chalcone (PKS3) and which are in general essential for CHS activity (PKS2). The possible functions of these residues are discussed. PMID- 11437246 TI - Activation of tomato PR and wound-related genes by a mutagenized tomato MAP kinase kinase through divergent pathways. AB - A mitogen-activated protein kinase kinase (MAPKK) gene, tMEK2, was isolated from tomato cv. Bonny Best. By mutagenesis, a permanently active variant, tMEK2MUT, was created. Both wild-type tMEK2 and mutant tMEK2MUT were driven by a newly described strong plant constitutive promoter, tCUP, in a tomato protoplast transient gene expression system. Pathogenesis-related genes, PRlb1, PR3 and Twi1, and a wound-inducible gene, ER5, were activated by tMEK2MUT. Specific inhibitors of p38 class MAPK inhibited tMEK2MUT-induced activation of PR3 and ER5 genes but not that of the PRlb1 or Twi1 gene. Arabidopsis dual-specificity protein tyrosine phosphatase 1 (DsPTP1) and maize protein phosphatase 1 (PP1) inhibited tMEK2MUT-induced activation of the ER5 gene and the Twi1 gene, respectively, whereas PRlb1 and PR3 were not affected by either AtDsPTP1, or maize PP1, or Arabidopsis protein phosphatase 2A (PP2A). We have demonstrated for the first time that a single MAPKK activates an array of PR and wound-related genes. Our observation indicates that the activation of the genes downstream of tMEK2 occurs through divergent pathways and that tMEK2 may play an important role in the interaction of signal transduction pathways that mediate responses to both biotic (e.g. disease) and abiotic stresses (e.g. wound responsiveness). PMID- 11437247 TI - Two large Arabidopsis thaliana gene families are homologous to the Brassica gene superfamily that encodes pollen coat proteins and the male component of the self incompatibility response. AB - The male component of the self-incompatibility response in Brassica has recently been shown to be encoded by the S locus cysteine-rich gene (SCR). SCR is related, at the sequence level, to the pollen coat protein (PCP) gene family whose members encode small, cysteine-rich proteins located in the proteo-lipidic surface layer (tryphine) of Brassica pollen grains. Here we show that the Arabidopsis genome includes two large gene families with homology to SCR and to the PCP gene family, respectively. These genes are poorly predicted by gene-identification algorithms and, with few exceptions, have been missed in previous annotations. Based on sequence comparison and an analysis of the expression patterns of several members of each family, we discuss the possible functions of these genes. In particular, we consider the possibility that SCR-related genes in Arabidopsis may encode ligands for the S gene family of receptor-like kinases in this species. PMID- 11437248 TI - Isolation and characterization of a Na+/H+ antiporter gene from the halophyte Atriplex gmelini. AB - With a homologous gene region we successfully isolated a Na+/H+ antiporter gene from a halophytic plant, Atriplex gmelini, and named it AgNHX1. The isolated cDNA is 2607 bp in length and contains one open reading frame, which comprises 555 amino acid residues with a predicted molecular mass of 61.9 kDa. The amino acid sequence of the AgNHX1 gene showed more than 75% identity with those of the previously isolated NHX1 genes from glycophytes, Arabidopsis thaliana and Oryza sativa. The migration pattern of AgNHX1 was shown to correlate with H+ pyrophosphatase and not with P-type H+-ATPase, suggesting the localization of AgNHX1 in a vacuolar membrane. Induction of the AgNHX1 gene was observed by salt stress at both mRNA and protein levels. The expression of the AgNHX1 gene in the yeast mutant, which lacks the vacuolar-type Na+/H+ antiporter gene (NHX1) and has poor viability under the high-salt conditions, showed partial complementation of the NHX1 functions. These results suggest the important role of the AgNHX1 products for salt tolerance. PMID- 11437249 TI - A leucine-rich repeat region is conserved in pollen extensin-like (Pex) proteins in monocots and dicots. AB - We previously isolated a pollen-specific gene encoding a pollen tube wall associated glycoprotein with a globular domain and an extensin domain from maize (mPex1). To evaluate which protein domains might be important for function, we isolated a second monocot gene (mPex2) and a dicot gene (tPex). Each gene encodes a signal sequence, an N-terminal globular domain comprised of a variable region, a leucine-rich repeat (LRR) with an adjacent cysteine-rich region, a transition region and an extensin-like C-terminal domain. The LRRs of the maize and tomato Pex proteins are highly conserved. Although the extensin domains in the maize and tomato proteins vary in length and in amino acid sequence, they are likely to be structurally conserved. Additional putative Pex gene sequences were identified by either GenBank search (Arabidopsis) or PCR (sorghum and potato): all encode conserved LRRs. The presence of a conserved LRR in the known and potential Pex proteins strongly suggests that this motif is involved in the binding of a specific ligand during pollen tube growth. Gene expression studies using RNA and protein blotting as well as promoter-reporter gene fusions in transient and stable transformation indicate that the tomato Pex gene is pollen-specific. PMID- 11437250 TI - Phenotypic changes in Arabidopsis caused by expression of a yeast vacuolar Ca2+/H+ antiporter. AB - In plants, cytosolic Ca2+ levels are tightly regulated, and changes in cytosolic Ca2+ have been implicated in converting numerous signals into adapted responses. Vacuolar ion transporters are thought to be key mediators of cytosolic Ca2+ concentrations. In an attempt to interpret the role of vacuolar Ca2+ transport in plant processes, we have expressed the yeast vacuolar Ca2+/H+ antiporter, VCX1, in Arabidopsis and tobacco. This transporter localizes to the plant vacuolar membrane. VCX1-expressing Arabidopsis plants displayed increased sensitivity to sodium and other ions. These ion sensitivities could be suppressed by addition of calcium to the media. VCX1-expressing plants demonstrated increased tonoplast enriched Ca2+/H+ antiport activity as well as increased Ca2+ accumulation. These results suggest that VCX1 expression in Arabidopsis could be a valuable tool with which to experimentally dissect the role of Ca2+ transport around the plant vacuole. PMID- 11437251 TI - Activation of defence-related genes during senescence: a correlation between gene expression and cellular damage. AB - The correlation between activation of defence-related gene expression and plant senescence was investigated by evaluating the presence of specific transcripts in various leaves of tobacco senescing plants. Expression of most genes examined was found to be induced shortly after flowering; however, each gene had its own characteristic timing of expression and level of RNA accumulation. Studies of the symptoms developed in senescing leaves responding to bacterial inoculation suggest that the accumulation of defence-related transcripts in these tissues might be related with the mechanism of senescence rather than with protection of the plant against pathogen infection. We observed that the high level of GUS expression directed by the beta-1,3-glucanase gn1 promoter, in senescing leaves of transgenic tobacco plants. decreased after bacterial inoculation, in correlation with the formation of symptoms. Reduction of gene expression was likely to be the reflection of the additional damage caused by the bacteria in the senescent tissues inoculated. PMID- 11437252 TI - Identification of a cytoskeleton-associated 120 kDa RNA-binding protein in developing rice seeds. AB - During rice seed development, prolamine RNAs are localized to the surface of the prolamine storage protein bodies (PBs), organelles bounded by the endoplasmic reticulum (ER). The exact mechanism by which prolamine RNAs are enriched on this ER subdomain is not known but recent evidence indicates the directed transport and targeting of prolamine RNAs to the prolamine PBs. As such a process involves RNA signal determinants and cytoskeleton-interacting proteins that recognize these signals, we obtained an enriched cytoskeleton-PB fraction and identified a prominent RNA-binding activity, Rp120, by RNA-binding UV-cross-linking assay. Recombinant cDNA clones of Rp120 revealed that the primary sequence shared considerable structural homology to the human transcriptional coactivator p100 and possessed a modular organization, four nucleic acid-binding SN domains, a tudor domain and a coil-coil domain. Consistent with the presence of SN domains, Rp120 binds a variety of RNAs including prolamine RNA. Interaction with the latter RNA, however, was specific as binding activity was evident only to the prolamine 3' UTR and not to the 5' UTR or coding sequences. Rp120 is also able to interact with other proteins as its sedimentation behavior in sucrose density gradient suggests an association with the cytoskeleton. The presence of a tudor domain, suggested to have a role in RNA processing or transport, together with the SN and coiled-coil domains are consistent with the view that Rp120 may be involved in RNA sorting in rice endosperm. PMID- 11437253 TI - Molecular and phenotypic specificity of an antisense PHYB gene in Arabidopsis. AB - The family of phytochrome photoreceptors plays an essential role in regulating plant growth and development in response to the light environment. An antisense PHYB transgene has been introduced into wild-type Arabidopsis and shown to inhibit expression of the PHYB sense mRNA and the phyB phytochrome protein 4- to 5-fold. This inhibition is specific to phyB in that the levels of the four other phytochromes, notably the closely related phyD and phyE phytochromes, are unaffected in the antisense lines. Antisense-induced reduction in phyB causes alterations of red light effects on seedling hypocotyl elongation, rosette leaf morphology, and chlorophyll content, similar to the phenotypic changes caused by phyB null mutations. However, unlike the phyB mutants, the antisense lines do not flower early compared to the wild type. Furthermore, unlike the phyB mutants, the antisense lines do not show a reduction in phyC level compared to the wild type, making it possible to unequivocally associate several of the photomorphogenic effects seen in phyB mutants with phytochrome B alone. These results indicate that an antisense transgene approach can be used to specifically inhibit the expression and activity of a single member of the phytochrome family and to alter aspects of shade avoidance responses in a targeted manner. PMID- 11437254 TI - A novel orfB-related gene of carrot mitochondrial genomes that is associated with homeotic cytoplasmic male sterility (CMS). AB - Three types of orfB-related genes, orfB-F1, orfB-F2 and orfB-CMS, were found in carrot mitochondrial genomes. OrfB-F1 has a structure similar to the reported orfB gene in sunflower, and orfB-F2 is a novel chimeric gene with about 200 bp of unknown sequence at the 3' end of the orfB-related sequence. All fertile plants examined contained either orfB-F1 or orfB-F2. OrfB-CMS is also a novel chimeric orfB-related gene with an additional 170 bp of unknown sequence at the 3' end. Fifteen carrot lines including petaloid CMS were examined by PCR analysis and all petaloid CMS plants were found to contain the orfB-CMS gene. The orfB-F2 and orfB CMS genes coexist in three lines, and these lines exhibit a CMS phenotype, suggesting that the CMS phenotype is associated with orfB-CMS and is independent of the presence of orfB-F2. Interestingly, differences in predicted amino acid sequence between orfB-CMS and orfB-F2 were very limited and restricted to the carboxy-terminal region of the protein. The orfB-related genes were transcribed as expected from their DNA structures, but orfB-CMS protein accumulated only in floral organs of the CMS lines. Four RNA editing sites were common to orfB-CMS and orfB-F2, and C-to-U editing occurred in both floral and leaf organs for orfB CMS. These results strongly suggest that the orfB-CMS gene is closely associated with the petaloid phenotype and its expression is not regulated by RNA editing, but by post-transcriptional events. PMID- 11437255 TI - The Rhizobium GstI protein reduces the NH4+ assimilation capacity of Rhizobium leguminosarum. AB - We show that the protein encoded by the glutamine synthetase translational inhibitor (gstI) gene reduces the NH4+ assimilation capacity of Rhizobium leguminosarum. In this organism, gstI expression is regulated by the ntr system, including the PII protein, as a function of the nitrogen (N) status of the cells. The GstI protein, when expressed from an inducible promoter, inhibits glutamine synthetase II (glnII) expression under all N conditions tested. The induction of gstI affects the growth of a glutamine synthetase I (glnA-) strain and a single amino acid substitution (W48D) results in the complete loss of GstI function. During symbiosis, gstI is expressed in young differentiating symbiosomes (SBs) but not in differentiated N2-fixing SBs. In young SBs, the PII protein modulates the transcription of NtrC-regulated genes such as gstI and glnII. The evidence presented herein strengthens the idea that the endocytosis of bacteria inside the cytoplasm of the host cells is a key step in the regulation of NH4+ metabolism. PMID- 11437256 TI - Heterologous expression of genes mediating enhanced fungal resistance in transgenic wheat. AB - Three cDNAs encoding the antifungal protein Ag-AFP from the fungus Aspergillus giganteus, a barley class II chitinase and a barley type I RIP, all regulated by the constitutive Ubiquitin1 promoter from maize, were expressed in transgenic wheat. In 17 wheat lines, stable integration and inheritance of one of the three transgenes has been demonstrated over four generations. The formation of powdery mildew (Erysiphe graminis f. sp. tritici) or leaf rust (Puccinia recondita f. sp. tritici) colonies was significantly reduced on leaves from afp or chitinase II- but not from rip I-expressing wheat lines compared with non-transgenic controls. The increased resistance of afp and chitinase II lines was dependent on the dose of fungal spores used for inoculation. Heterologous expression of the fungal afp gene and the barley chitinase II gene in wheat demonstrated that colony formation and, thereby, spreading of two important biotrophic fungal diseases is inhibited approximately 40 to 50% at an inoculum density of 80 to 100 spores per cm2. PMID- 11437257 TI - Cell biological changes of outer cortical root cells in early determinate nodulation. AB - In the symbiosis of leguminous plants and Rhizobium bacteria, nodule primordia develop in the root cortex. This can be either in the inner cortex (indeterminate type of nodulation) or outer cortex (determinate-type of nodulation), depending upon the host plant. We studied and compared early nodulation stages in common bean (Phaseolus vulgaris) and Lotus japonicus, both known as determinate-type nodulation plants. Special attention was paid to the occurrence of cytoplasmic bridges, the influence of rhizobial Nod factors (lipochitin oligosaccharides [LCOs]) on this phenomenon, and sensitivity of the nodulation process to ethylene. Our results show that i) both plant species form initially broad, matrix-rich infection threads; ii) cytoplasmic bridges occur in L. japonicus but not in bean; iii) formation of these bridges is induced by rhizobial LCOs; iv) formation of primordia starts in L. japonicus in the middle root cortex and in bean in the outer root cortex; and v) in the presence of the ethylene biosynthesis inhibitor aminoethoxyvinylglycine (AVG), nodulation of L. japonicus is stimulated when the roots are grown in the light, which is consistent with the role of cytoplasmic bridges during nodulation of L. japonicus. PMID- 11437258 TI - Responses of a model legume Lotus japonicus to lipochitin oligosaccharide nodulation factors purified from Mesorhizobium loti JRL501. AB - Lotus japonicus has been proposed as a model legume for molecular genetic studies of symbiotic plant-microbe interactions leading to the fixation of atmospheric nitrogen. Lipochitin oligosaccharides (LCOs), or Nod factors, were isolated from the culture of Mesorhizobium loti strain JRL501 (MAFF303099), an efficient microsymbiont of L. japonicus B-129 cv. Gifu. High-performance liquid chromatography and mass spectrometric analyses allowed us to identify at least five different structures of LCOs that were produced by JRL501. The major component was NodMl-V(C18:1, Me, Cb, AcFuc), an N-acetyl-glucosamine pentamer in which the nonreducing residue is N-acylated with a C18:1 acyl moiety, N methylated, and carries a carbamoyl group and the reducing N-acetylglucosamine residue is substituted with 4-O-acetyl-fucose. Additional novel LCO structures bearing fucose instead of acetyl-fucose at the reducing end were identified. Mixtures of these LCOs could elicit abundant root hair deformation on L. japonicus roots at a concentration of 10(-7) to 10(-9) M. Spot inoculation of a few nanograms of LCOs on L. japonicus roots induced the formation of nodule primordia in which the early nodulin genes, ENOD40 and ENOD2, were expressed in a tissue-specific manner. We also observed the formation of a cytoplasmic bridge (preinfection thread) in the swollen outermost cortical cells. This is the first description of cytoplasmic bridge formation by purified LCOs alone in a legume forming determinate nodules. PMID- 11437259 TI - Knockout of an azorhizobial dTDP-L-rhamnose synthase affects lipopolysaccharide and extracellular polysaccharide production and disables symbiosis with Sesbania rostrata. AB - A nonpolar mutation was made in the oac2 gene of Azorhizobium caulinodans. oac2 is an ortholog of the Salmonella typhimurium rfbD gene that encodes a dTDP-L rhamnose synthase. The knockout of oac2 changed the lipopolysaccharide (LPS) pattern and affected the extracellular polysaccharide production but had no effect on bacterial hydrophobicity. Upon hot phenol extraction, the wild-type LPS partitioned in the phenol phase. The LPS fraction of ORS571-oac2 partitioned in the water phase and had a reduced rhamnose content and truncated LPS molecules on the basis of faster migration in detergent gel electrophoresis. Strain ORS571 oac2 induced ineffective nodule-like structures on Sesbania rostrata. There was no clear demarcation between central and peripheral tissues, and neither leghemoglobin nor bacteroids were present. Light and electron microscopy revealed that the mutant bacteria were retained in enlarged, thick-walled infection threads. Infection centers emitted a blue autofluorescence under UV light. The data indicate that rhamnose synthesis is important for the production of surface carbohydrates that are required to sustain the compatible interaction between A. caulinodans and S. rostrata. PMID- 11437260 TI - No evidence for binding between resistance gene product Cf-9 of tomato and avirulence gene product AVR9 of Cladosporium fulvum. AB - The gene-for-gene model postulates that for every gene determining resistance in the host plant, there is a corresponding gene conditioning avirulence in the pathogen. On the basis of this relationship, products of resistance (R) genes and matching avirulence (Avr) genes are predicted to interact. Here, we report on binding studies between the R gene product Cf-9 of tomato and the Avr gene product AVR9 of the pathogenic fungus Cladosporium fulvum. Because a high affinity binding site (HABS) for AVR9 is present in tomato lines, with or without the Cf-9 resistance gene, as well as in other solanaceous plants, the Cf-9 protein was produced in COS and insect cells in order to perform binding studies in the absence of the HABS. Binding studies with radio-labeled AVR9 were performed with Cf-9-producing COS and insect cells and with membrane preparations of such cells. Furthermore, the Cf-9 gene was introduced in tobacco, which is known to be able to produce a functional Cf-9 protein. Binding of AVR9 to Cf-9 protein produced in tobacco was studied employing surface plasmon resonance and surface-enhanced laser desorption and ionization. Specific binding between Cf-9 and AVR9 was not detected with any of the procedures. The implications of this observation are discussed. PMID- 11437261 TI - Characterization of a PR-10 pathogenesis-related gene family induced in rice during infection with Magnaporthe grisea. AB - A partial cDNA with homology to the PR-10 class of pathogenesis-related proteins was used to screen a rice genomic library. One 16-kb genomic clone contained three genes with PR-10 similarity. These genes, RPR10a, RPR10b, and RPR10c, were arranged in tandem and separated by approximately 2.5 kb. RPR10a cDNA was obtained by reverse transcription-polymerase chain reaction, and sequence analysis revealed that RPR10a and RPR10b encode predicted proteins of 158 and 160 amino acids, respectively, and share 71% amino acid identity. RPR10c appears to be a nonfunctional pseudogene. Gene-specific probes were used to study transcript accumulations of the three RPR10 genes in rice plants following inoculation with Magnaporthe grisea. RPR10a transcripts were induced from a low basal level within 12 h after inoculation and showed a second higher level induction at 48 h, which continued throughout the 144 h it was examined. In addition, RPR10a was induced strongly by salicylic and jasmonic acid applications to rice plants. Transcripts of RPR10b also were enhanced by M. grisea, but were not strongly visible until 48 h after inoculation. Tissue prints of M. grisea-infected rice leaves when the RPR10a-specific probe was used indicate that RPR10a is expressed most strongly in a localized fashion in response to the pathogen. PMID- 11437262 TI - Mutation in the ntrR gene, a member of the vap gene family, increases the symbiotic efficiency of Sinorhizobium meliloti. AB - In specific plant organs, namely the root nodules of alfalfa, fixed nitrogen (ammonia) produced by the symbiotic partner Sinorhizobium meliloti supports the growth of the host plant in nitrogen-depleted environment. Here, we report that a derivative of S. meliloti carrying a mutation in the chromosomal ntrR gene induced nodules with enhanced nitrogen fixation capacity, resulting in an increased dry weight and nitrogen content of alfalfa. The efficient nitrogen fixation is a result of the higher expression level of the nifH gene, encoding one of the subunits of the nitrogenase enzyme, and nifA, the transcriptional regulator of the nif operon. The ntrR gene, controlled negatively by its own product and positively by the symbiotic regulator syrM, is expressed in the same zone of nodules as the nif genes. As a result of the nitrogen-tolerant phenotype of the strain, the beneficial effect of the mutation on efficiency is not abolished in the presence of the exogenous nitrogen source. The ntrR mutant is highly competitive in nodule occupancy compared with the wild-type strain. Sequence analysis of the mutant region revealed a new cluster of genes, termed the "ntrPR operon," which is highly homologous to a group of vap-related genes of various pathogenic bacteria that are presumably implicated in bacterium-host interactions. On the basis of its favorable properties, the strain is a good candidate for future agricultural utilization. PMID- 11437263 TI - A dysfunctional movement protein of tobacco mosaic virus interferes with targeting of wild-type movement protein to microtubules. AB - The Tobacco mosaic virus (TMV) movement protein (MPTMV) mediates cell-to-cell viral trafficking by altering properties of the plasmodesmata (Pd) in infected cells. During the infection cycle, MPTMV becomes transiently associated with endomembranes, microfilaments, and microtubules (MT). It has been shown that the cell-to-cell spread of TMV is reduced in plants expressing the dysfunctional MP mutant MPNT-1. To expand our understanding of the MP function, we analyzed events occurring during the intracellular and intercellular targeting of MPTMV and MPNT 1 when expressed as a fusion protein to green fluorescent protein (GFP), either by biolistic bombardment in a viral-free system or from a recombinant virus. The accumulation of MPTMV:GFP, when expressed in a viral-free system, is similar to MPTMV:GFP in TMV-infected tissues. Pd localization and cell-to-cell spread are late events, occurring only after accumulation of MP:GFP in aggregate bodies and on MT in the target cell. MPNT-1:GFP localizes to MT but does not target to Pd nor does it move cell to cell. The spread of transiently expressed MPTMV:GFP in leaves of transgenic plants that produce MPNT-1 is reduced, and targeting of the MPTMV:GFP to the cytoskeleton is inhibited. Although MPTMV:GFP targets to the Pd in these plants, it is partially impaired for movement. It has been suggested that MPNT-1 interferes with host-dependent processes that occur during the intracellular targeting program that makes MP movement competent. PMID- 11437264 TI - Possible involvement of the phloem lectin in long-distance viroid movement. AB - Incubation with cucumber phloem exudate in vitro results in a dramatic decrease in the electrophoretic mobility of Hop stunt viroid. UV cross-linking and a combination of size exclusion and ion exchange chromatography indicate that this phenomenon reflects a previously unsuspected ability of phloem protein 2, a dimeric lectin and the most abundant component of phloem exudate, to interact with RNA. In light of its demonstrated ability to move from cell to cell via plasmodesmata as well as long distances in the phloem, our results suggest that phloem protein 2 may facilitate the systemic movement of viroids and, possibly, other RNAs in vivo. PMID- 11437265 TI - Identification of an in vitro ribonucleoprotein complex between a viroid RNA and a phloem protein from cucumber plants. AB - We used the interaction of Hop stunt viroid (HSVd) and cucumber plants to investigate the involvement of phloem proteins in the systemic transport of RNA molecules. A ribonucleoprotein complex, stable even at high salt and temperature conditions, was detected in vitro between HSVd-RNA and the phloem exudate obtained from sectioned internodes from cucumber plants. The phloem protein 2 was recovered from this ribonucleoprotein complex and its RNA-binding properties as demonstrated by gel retardation analysis. The involvement of this protein in the movement of RNAs in cucumber is discussed. PMID- 11437266 TI - Mutants of tobacco mosaic virus with temperature-sensitive coat proteins induce heat shock response in tobacco leaves. AB - We analyzed, with respect to heat shock proteins (HSPs), systemically reacting tobacco leaves inoculated with Tobacco mosaic virus (TMV), wild-type vulgare, and temperature-sensitive coat protein (CP) mutants Ni 118 (P20L) and flavum (D19A), kept at 23 or 30 degrees C. HSP18 and HSP70 mRNAs and proteins were induced with temperature-sensitive CP mutants after 1 to 2 days at 30 degrees C. After 4 to 6 days, HSP70 was also induced at 23 degrees C. The induction of HSPs paralleled the amount of insoluble TMV CP in leaf extracts, indicating that denatured TMV CP by itself induces a heat-shock response. PMID- 11437267 TI - Analysis of a symbiosis-specific cytochrome P450 homolog in Rhizobium sp. BR816. AB - Sequence analysis of the DNA region upstream of nodO in Rhizobium sp. BR816 revealed an open reading frame in which the deduced amino acid sequence shows homology with cytochrome P450. Because the BR816 P450 homolog shows 73% amino acid similarity with CYP127A1(Y4vG), which is identified on the symbiotic plasmid of Rhizobium sp. NGR234, it is named CYP127A2. Transcriptional analysis of CYP127A2 revealed high expression in bacteroids, whereas no or hardly any expression was observed under free-living conditions. Low-level, free-living expression, however was noticed when cells were grown microoxically at acid pH levels. A number of possible substrates that may induce P450 gene expression were analyzed, but only the addition of short-chain alcohols to cultures slightly increased CYP127A2 expression. High levels of CYP127A2 expression observed in bacteroids of a nifH mutant strain, which formed non-fixing nodules on bean, indicated that the genuine substrate for CYP127A2 is not a metabolite resulting from N2-fixation. Nevertheless, expression analysis pointed to a NifA- and sigma54-dependent transcription. PMID- 11437268 TI - Effect of fragarin on the cytoplasmic membrane of the phytopathogen Clavibacter michiganensis. AB - Fragarin, an antibiotic that was isolated and purified from a soluble fraction of strawberry leaves, may be a new type of preformed antimicrobial compound (phytoanticipin). Here, we report that the growth and oxygen consumption of the phytopathogenic bacterium Clavibacter michiganensis were rapidly inhibited after the addition of fragarin to cultures. Also, dissipation of the membrane potential and an increase of cell membrane permeability were observed in the presence of fragarin. The ability of fragarin to dissipate the membrane potential was confirmed with the use of small unilamellar liposomes made with lipids extracted from C. michiganensis. Our results suggest that fragarin is able to act at the membrane level, and that this action is correlated with a decrease in cell viability. PMID- 11437269 TI - Knowledge, authority and identity: a prolegomenon to an epistemology of the clinic. AB - Disputes about theory in bioethics almost invariably revolve around different understandings of morality or practical reasoning; I here suggest that the field would do well to become more explicitly contentious about knowledge, and start the task of putting together a clinical epistemology. By way of providing some motivation for such a discussion, I consider two cases of resistance to shifts in clinical practice that are, by and large, not ethically controversial, highlighting how different conceptions of epistemic authority may contribute to clinicians' unwillingness to adopt these changes, and sketching out some initial suggestions for epistemic analysis of clinical practice. PMID- 11437270 TI - Theory and the organic bioethicist. AB - This article argues for the importance of theoretical reflections that originate from patients' experiences. Traditionally academic philosophers have linked their ability to theorize about the moral basis of medical practice to their role as outside observer. The author contends that recently a new type of reflection has come from within particular patient populations. Drawing upon a distinction created by Antonio Gramsci, it is argued that one can distinguish the theory generated by traditional bioethicists, who are academically trained, from that of "organic" bioethicists, who identify themselves with a particular patient community. The characteristics of this new type of bioethicist that are explored in this article include a close association of memoir and philosophy, an interrelationship of theory and praxis, and an intimate connection between the individual and a particular patient community. PMID- 11437271 TI - The epistemology of communitarian bioethics: traditions in the public debates. AB - I consider the problem liberalism poses for bioethics. Liberalism is a view that advocates that the state remain neutral to views of the good life. This view is sometimes supported by a skeptical moral epistemology that tends to propel liberalism toward libertarianism. I argue that the possibilities for shared agreement on moral matters are more promising than is sometimes appreciated by such a view of liberalism. Using two examples of public debates of moral issues, I show that commonly shared intuitions may ground moral principles even if they may be given different weight by persons of different moral and religious traditions. Nevertheless, the fact that the intuition and principle is widely shared may be sufficient to chart some directions for public policy or cooperative action even if they do not lead to complete agreement. As a result, I argue that a liberal communitarianism that presupposes a fairly minimalist epistemology is a legitimate approach to achieving shared agreement in a pluralistic society. PMID- 11437272 TI - Theory and engagement in bioethics. PMID- 11437273 TI - Freestanding pragmatism in law and bioethics. AB - This paper represents the first installment of a larger project devoted to the relevance of pragmatism for bioethics. One self-consciously pragmatist move would be to return to the classical pragmatist canon of Peirce, James and Dewey in search of substantive doctrines or methodological approaches that might be applied to current bioethical controversies. Another pragmatist (or neopragmatist) move would be to subject the regnant principlist paradigm to Richard Rorty's subversive assaults on foundationalism in epistemology and ethics. A third pragmatist method, dubbed "freestanding pragmatism" by its proponents, embraces a "pragmatist" approach to practical reasoning without discernable moorings either to the classical canon or to Rorty's neopragmatism. This third pragmatist approach to method in practical ethics is the subject of this article. I begin with an examination of freestanding pragmatism in the theory of judicial decision making. I argue that this version of legal pragmatism -so described on account of its commitments to contextualism, instrumentalism, eclecticism, and freedom from grand theory--bears a striking resemblance to much self-described pragmatist work in bioethics today. I further argue that if this is what we mean by "pragmatism," then in a certain sense "we are all pragmatists now." PMID- 11437274 TI - The independence of practical ethics. AB - After criticizing three common conceptions of the relationship between practical ethics and ethical theory, an alternative modeled on Aristotle's conception of the relationship between rhetoric and philosophical ethics is explored. This account is unique in that it neither denigrates the project of searching for an adequate comprehensive ethical theory nor subordinates practical ethics to that project. Because the purpose of practical ethics, on this view, is to secure the cooperation of other persons in a way that respects their status as free and equal, it seeks to influence the judgments of others by providing them with reasons that are accessible to their own understanding. On this account, the independence of practical ethics is rooted in an appreciation of the constraints that nonideal circumstances place on the role that the philosophically refined premises of moral theory can play in such public deliberations. Practical and philosophical ethics are united, not by shared theoretical frameworks or principles, but by the need to exercise intelligently the same intellectual and affective capacities. They are separated, not by the particularity or generality of their starting points, but by their responsiveness to the practical problem of facilitating sound normative deliberations among persons as we find them, under non-ideal circumstances. PMID- 11437275 TI - Use of epitope tags for routine analysis of transgene expression. AB - Peptide and RNA epitope tags as tools for routine analysis of transgene expression and protein accumulation in transformed plant cell cultures was evaluated using three genes that encode very structurally and functionally different proteins. A T7 peptide was introduced at the amino- and carboxyl termini of phosphinothricin-N-acetyl transferase and avidin and at the carboxyl terminus of galactose oxidase. An RNA sequence that forms a higher order structure that is recognized by antibodies raised against the FLAG peptide was separately introduced into the 3' nontranslated region of these genes. Constructs were introduced into maize cell cultures using particle bombardment and transgene expression, protein accumulation, protein function and presence of the tags in RNA and/or protein as appropriate were evaluated in up to approximately 25 culture lines per construct. Results indicate that, while there will likely always be a need for some empirical evaluation of any tag-protein combination, introduction of the peptide tag at the amino-terminus was generally more successful than was incorporation at the carboxyl-terminus. RNA tags show promise for this purpose, but routine application will require development of a very sensitive immunoassay. PMID- 11437276 TI - Expression of recombinant human granulocyte macrophage-colony stimulating factor (hGM-CSF) in mouse urine. AB - We have generated transgenic mice expressing human granulocyte macrophage-colony stimulating factor (hGM-CSF) in urine. In particular, the expression plasmid DNA containing mouse uroplakin II promoter was used to direct uroepithelium-specific transcription of transgene. In this study, hGM-CSF transcript was detected only in bladder uroepithelium as determined by northern blot analysis. Furthermore, hGM-CSF protein was detected in the suprabasal layer of the uroepithelium and ureter by immunohistochemistry. The hGM-CSF was secreted into urine at high level (up to 180 ng/ml), and enhanced proliferation of hGM-CSF-dependent human acute monocyte leukemic cells, suggesting that transgenic urine-derived hGM-CSF was bioactive. This is the first case of demonstrating biological activity of a cytokine produced in the urine of a transgenic animal. Our results demonstrate that bladder can be used as a bioreactor to produce biologically important substances. In addition, it suggests a potential application of bladder expression system to livestock for high-yield production of pharmaceuticals. PMID- 11437277 TI - Post-transcriptional gene silencing in plum pox virus resistant transgenic European plum containing the plum pox potyvirus coat protein gene. AB - Transgenic plums containing the plum pox potyvirus coat protein (PPV-CP) gene were inoculated with PPV. Infection was monitored by evaluating symptoms, ELISA, and IC-RT-PCR. Transgenic clone C5 was highly resistant to PPV during four years of testing and displayed characteristics typical of post-transcriptional gene silencing (PTGS), including a high level of transgene transcription in the nucleus, low levels of transgene mRNA in the cytoplasm, a complex multicopy transgene insertion with aberrant copies, and methylation of the silenced PPV-CP transgene. The PPV-CP transgene was also methylated in seedlings of C5 and these seedlings were resistant to PPV. Our results show, for the first time, that PTGS functions as a mechanism for virus resistance in a woody perennial species. PMID- 11437278 TI - Effective generation of very low density lipoprotein receptor transgenic mice by overlapping genomic DNA fragments: high testis expression and disturbed spermatogenesis. AB - The generation of functional transgenes via microinjection of overlapping DNA fragments has previously been reported to be successful, but it is still not a widely applied approach. Here we show that the method is very reliable, and should be considered, in case a single large insert clone of the desired gene is not available. In the present study, two large DNA fragments consisting of overlapping cosmids, together constituting the human very low density lipoprotein receptor (VLDLR) gene (35 kb), were used to generate VLDLR transgenic (VLDLR-Tg) mice. Three transgenic founders were born, of which two (strain #2 and #3) generated transgenic offspring. Using Fiber-FISH analysis, the integration site was shown to contain at least 44 and 64 DNA fragments in mouse strains #2 and #3, respectively. This copy number resulted in integration sites of 1.5 and 2.5 megabase in size. Notably, over 90% of the fragments in both mouse strains #2 and #3 were flanked by their complementary fragment. In line with this observation, Southern blot analysis demonstrated that the correct recombination between fragments predominated in the transgenic insertion. Human VLDLR expression was detected in testis, kidney and brain of both mouse strains. Since this pattern did not parallel the endogenous VLDLR expression, some crucial regulatory elements were probably not present in the cosmid clones. Human VLDLR expression in testis was detected in germ cells up to the meiotic stage by in situ mRNA analysis. Remarkably, in the F1 generation of both VLDLR-Tg mouse strains the testis was atrophic and giant cells were detected in the semineferous tubuli. Furthermore, male VLDLR-Tg mice transmitted the transgene to their progeny with low frequencies. This could imply that VLDLR overexpression in the germ cells disturbed spermatogenesis. PMID- 11437279 TI - Influence of plant development and environment on transgene expression in potato and consequences for insect resistance. AB - Clonal replicates of different transformed potato plants expressing transgene constructs containing the constitutive Cauliflower Mosaic Virus (CaMV) 35S promoter, and sequences encoding the plant defensive proteins snowdrop lectin (Galanthus nivalis agglutinin; GNA), and bean chitinase (BCH) were propagated in tissue culture. Plants were grown to maturity, at first under controlled environmental conditions, and later in the glasshouse. For a given transgene product, protein accumulation was found to vary between the different lines of clonal replicates (where each line was derived from a single primary transformant plant), as expected. However, variability was also found to exist within each line of clonal replicates, comparable to the variation of mean expression levels observed between the different clonal lines. Levels of GNA, accumulated in different parts of a transgenic potato plant, also showed variation but to a lesser extent than plant-plant variation in expression. With the majority of the clonal lines investigated, accumulation of the transgene product was found to increase as the potato plant developed, with maximum levels found in mature plants. The variation in accumulation of GNA among transgenic plants within a line of clonal replicates was exploited to demonstrate that the enhanced resistance towards larvae of the tomato moth, Lacanobia oleracea L., caused by expression of this protein in potato, was directly correlated with the level of GNA present in the plants, and that conditions under which the plants were grown affect the levels of GNA expression and subsequent levels of insect resistance. PMID- 11437280 TI - Molecular characterization of transgenic shallots (Allium cepa L.) by adaptor ligation PCR (AL-PCR) and sequencing of genomic DNA flanking T-DNA borders. AB - Genomic DNA blot hybridization is traditionally used to demonstrate that, via genetic transformation, foreign genes are integrated into host genomes. However, in large genome species, such as Allium cepa L., the use of genomic DNA blot hybridization is pushed towards its limits, because a considerable quantity of DNA is needed to obtain enough genome copies for a clear hybridization pattern. Furthermore, genomic DNA blot hybridization is a time-consuming method. Adaptor ligation PCR (AL-PCR) of genomic DNA flanking T-DNA borders does not have these drawbacks and seems to be an adequate alternative to genomic DNA blot hybridization. Using AL-PCR we proved that T-DNA was integrated into the A. cepa genome of three transgenic lines transformed with Agrobacterium tumefaciens EHA 105 (pCAMBIA 1301). The AL-PCR patterns obtained were specific and reproducible for a given transgenic line. The results showed that T-DNA integration took place and gave insight in the number of T-DNA copies present. Comparison of AL-PCR and previously obtained genomic DNA blot hybridization results pointed towards complex T-DNA integration patterns in some of the transgenic plants. After cloning and sequencing the AL-PCR products, the junctions between plant genomic DNA and the T-DNA insert could be analysed in great detail. For example it was shown that upon T-DNA integration a 66 bp genomic sequence was deleted, and no filler DNA was inserted. Primers located within the left and right flanking genomic DNA in transgenic shallot plants were used to recover the target site of T-DNA integration. PMID- 11437281 TI - Tet-system for the regulation of gene expression during embryonic development. AB - The ability to control gene expression in a temporal and spatial manner provides a new tool for the study of mammalian gene function particularly during development and oncogenesis. In this study the suitability of the tet-system for investigating embryogenesis was tested in detail. The tTACMV(M1) and rTACMV-3 (reverse Tc-controlled transactivator) transgenic mice were bred with NZL-2 bi reporter mice containing the vector with a tTA/rTA responsive bidirectional promoter that allows simultaneous regulation of expression of two reporter genes encoding luciferase and beta-galactosidase. In both cases reporter genes were found to be expressed in a wide spectrum of tissues of double transgenic embryos and adult mice. The earliest expression was detected in tTACMV(M1)/NZL-2 embryos at embryonic day 10.5 (E10.5) and rTACMV-3/NZL-2 embryos at E13.5. Doxycycline abolished beta-gal expression in tTACMV(M1)/NZL-2 but induced it in rTACMV-3/NZL 2 embryos including late stages of embryo-genesis. The tTA and rtTA transactivators thus revealed a partially complementary mode of action during second half of embryonic development. These experiments demonstrated that both Tet regulatory systems function during embryonic development. We conclude that the Tet systems allows regulation of gene expression during embryonic development and that 'double reporter' animals like the NZL-2 mice are useful tools for the characterization of newly generated tet transactivator lines expressing tTA (or rtTA) in embryonic as well as in adult tissues. PMID- 11437282 TI - Germ-line transformation of Arabidopsis lasiocarpa. AB - In planta transformation methods have opened up the possibility of transforming plant species for which no regeneration protocols currently exist. In this study, the suitability of the germ-line transformation method developed for Arabidopsis thaliana was examined for four taxa in the Brassicaceae that have not been previously transformed: Arabidopsis griffithiana, Arabidopsis lasiocarpa, Arabidopsis petraea and Capsella bursa-pastoris. Numerous transformants were obtained for A. lasiocarpa. Transformation of A. lasiocarpa was confirmed at the phenotypic and molecular levels for stably transformed lines and for backcrossed lines segregating the T-DNA insert. Parameters affecting transformation efficiency of A. lasiocarpa were also explored. As with A. thaliana, sucrose and surfactant in the inoculation medium are required for high levels of transformation, although the suitable concentrations of these are different for A. lasiocarpa. Other components present in earlier versions of the inoculation medium had little effect on transformation efficiency. Vacuum infiltration (rather than simple floral dipping) led to higher rates of transformation and did not seriously affect seed production in A. lasiocarpa. Identification of species susceptible to germ-line transformation will aid in determining the factors important for applying this technology to more recalcitrant species. PMID- 11437283 TI - Codon optimization markedly improves doxycycline regulated gene expression in the mouse heart. AB - Tetracycline regulated gene expression in transgenic animals is potentially a very powerful technique (Furth et al., 1994; Gossen & Bujard 1992). We have utilized this system in an attempt to overcome the perinatal lethality resulting from constitutive transgenic expression in the heart (Valencik & McDonald, Am J Physiol Heart Circ Physiol 280: H361-H367). We found that compound hemizygous animals created by mating selected reverse tetracycline transactivator (rtTA) and transresponder (TR) lines display tightly regulated TR expression in the heart. However, we identified two fundamental problems. First, codon usage bias appeared to severely limit the expression of the rtTA driven by the cardiac alpha-myosin heavy chain promoter. Second, co-injection of rtTA and TR transgenes led to compound hemizygous animals that exhibited unregulated TR gene expression. Codon optimization of the rtTA construct leads to marked improvement (increasing the average induction from 20-fold to 832-fold) in cardiac myocyte expression. The resulting opt-rtTA lines can be bred to homozygosity, facilitating rapid screening of F0 TR animals for doxycycline regulated transgene expression. PMID- 11437284 TI - Influences of venous involvement on postoperative brain damage following the anterior interhemispheric approach. AB - BACKGROUND: The anterior interhemispheric approach offers us an excellent surgical view for suprasellar lesions. Following this approach, we occasionally encounter postoperative brain damage in the frontal lobes. To assess the determinants of such a complication, we undertook a clinical study. METHOD: Potential causes for such brain damage were evaluated in 28 consecutive patients with suprasellar tumours extirpated using this approach. We focused particularly on the influences of venous involvement during surgery. The draining territory index (DTI) was originally devised for estimating the extent of the draining area of the sacrificed bridging veins. FINDINGS: CT evident brain damage was observed in five of 28 patients (17.8%), but only one patient (3.6%) showed clinically significant postoperative deficits. The patient's age, tumour pathology, tumour character, tumour size, duration of surgery, and radicality of the surgery did not affect the incidence of the brain damage. Of the twelve patients whose bridging veins were sacrificed during surgery, four (33.3%) showed brain damage in the frontal lobes. In contrast, such damage was observed in only one patient out of 16 (6.3%) whose bridging veins were preserved. Among the brain-damaged group, the average DTI of the sacrificed veins was significantly higher than that among the non-brain-damaged group. INTERPRETATION: Venous involvement during surgery significantly aggravated postoperative brain damage following the anterior interhemispheric approach. The DTI was useful in predicting the risk of brain damage, and a large bridging vein with a DTI over 50% should not be sacrificed during surgery. PMID- 11437285 TI - Open MRI-guided microsurgery of intracranial tumours in or near eloquent brain areas. AB - OBJECTIVES: Preservation of brain function while maximizing resection is the main aim of brain tumour surgery. The purpose of this study was to evaluate the efficacy of intra-operative magnetic resonance imaging to preserve brain function in patients with tumours in or near eloquent brain areas. METHODS AND MATERIAL: Between August 1997 and January 1999 fifty-eight craniotomies for intracranial tumours or vascular malformations have been performed at the University of Leipzig using a 0.5 T superconducting MR system "SIGNA SP" (General Electric Medical Systems, USA). In 32 of these patients (15 male/17 female) with intracranial tumours, located in or near eloquent brain areas (sensorimotor cortex/speech center), 34 craniotomies were performed using the image guidance of the interventional MRI. RESULTS: Using intra-operative MRI criteria, complete tumour removal could be achieved in 28 (82%) of 34 procedures. In 3 patients only subtotal tumour removal was possible, because the residual tumour was not visible on the intra-operative MR images, but could be identified on early diagnostic follow-up MR-scans. In 3 patients, incomplete tumour resection was performed in order to avoid neurological impairment. In these patients intra-operative MR images revealed residual tissue abnormalities involving or encroaching on deep brain structures or motor/language cortex. Pre-operative neurological status was unchanged in 24 patients (70%), worsened in 4 patients (12%) and improved in 6 patients (18%). CONCLUSIONS: Intra-operative MRI is helpful for navigation as well as to demonstrate the tumour margins to achieve a complete and safe resection of intracranial lesions located in or near eloquent brain areas. It enables an image based functional monitoring of the brain which is critical for motor, sensory or language function. Complications related to the surgical procedure are reduced and the risk of neurological deterioration due to tumour removal and postoperative complications is minimized. PMID- 11437286 TI - Congestive brain oedema associated with a pial arteriovenous malformation with impaired venous drainage. AB - We describe two patients with an unruptured pial AVM accompanied by significant brain oedema at initial presentation. In both cases, the primary drainer was a cortical vein showing varicose dilatation. in which venous congestion was indicated by magnetic resonance imaging (MRI). The restriction of venous drainage presumably caused venous hypertension in the surrounding brain, leading to the brain oedema and neurological symptoms. Brain oedema can develop in patients with an unruptured AVM by venous congestion following spontaneous thrombosis of venous components. Varicosity in a major cortical draining vein and a small nidus are the possible lesions predisposing this fairly rare condition for unruptured AVMs. PMID- 11437287 TI - Management-related morbidity and mortality in unselected aneurysms of the basilar trunk and vertebrobasilar junction. AB - OBJECTS: To analyze the management-related morbidity and mortality in unselected aneurysms of the basilar trunk and vertebrobasilar junction. The secondary objective was to investigate the factors associated with favourable or unfavourable surgical outcome in order to define subgroups for surgical and endovascular treatment. METHODS: 24 consecutive patients with aneurysms of the basilar trunk and vertebrobasilar junction were included in this study. They comprised 2.7% of all aneurysms treated during the study period between 1990 and 1997. 22 patients presented with acute subarachnoid hemorrhage (SAH) and 2 patients with symptoms of brainstem compresssion. All patients were managed using a standard protocol including surgery at the earliest possible moment, aggressive tripe-H therapy in patients with symptomatic vasospasm and mandatory follow-up angiography. 23 patients underwent surgical clipping and one patient endovascular coiling of the aneurysm. 12 patients had an excellent outcome. 6 patients had a good outcome, resulting in a total of satisfactory outcomes in 18 patients (75%). 4 patients (17%) had moderate to severe deficits. Two patients died (8%). Both patients had fusiform basilar trunk aneurysms. Good or excellent outcome was observed in 7 of 8 patients with aneurysms of the vertebrobasilar junction, 13 of 14 patients with moderate or minor SAH or without SAH (Fisher grade 0 to 2) and all patients with small sized aneurysm (n = 6). Factors mostly associated with poor outcome or death after surgical treatment were aneurysm location at the basilar trunk, large aneurysm size or fusiforme aneurysm type and severe SAH. CONCLUSIONS: Location, aneurysm size and the severity of SAH may help to predict the subgroup which highly benefits from surgical clipping of these rare vascular lesions. PMID- 11437288 TI - Endovascular treatment of radiation-induced petrous internal carotid artery aneurysm presenting with acute haemorrhage. A report of two cases. AB - Haemorrhage from rupture of petrous ICA aneurysm can be life threatening and emergency treatment is required. We report 2 cases of radiation-induced petrous internal carotid artery (ICA) aneurysm presenting with acute haemorrhage (epistaxis and otorrhagia) after radiotherapy (RT) for nasopharyngeal carcinoma (NPC). Both patients had a history of RT treatment for NPC. The first patient, a 54-year-old man, presented with sudden severe epistaxis and haemorrhagic shock. The second patient, a 35-year-old man, presented with episodes of severe otorrhagia. The first patient was immediately resuscitated. Obliteration of the aneurysm was performed by endovascular occlusion of the ICA with Guglielmi detachable coils and fibered platinum coils. For the second patient, the aneurysm was treated by deploying a self-expandable stent across the aneurysm neck. In an emergency situation, ruptured petrous ICA aneurysm can be treated with endovascular occlusion of the ICA with microcoils if there is a good collateral blood flow. Alternatively, the aneurysm can be treated by deployment of a stent, which can induce stasis and eventual thrombosis of the aneurysm. PMID- 11437289 TI - Malignant peripheral nerve sheath tumours--report of 8 cases and review of the literature. AB - BACKGROUND: Though Malignant peripheral nerve sheath tumours (MPNST) are a rare entity accounting for 5-10% of soft-tissue sarcomas they are an important differential diagnosis to benign tumours of the peripheral nervous system regarding treatment and prognosis. METHOD: We present our experience with eight patients who underwent surgery for MPNST at the Department of Neurosurgery between 10/1990 and 9/1999. The median age was 37 years [range: 13-64], the male/female ratio was 1:1. Two patients suffered from Neurofibromatosis type 1. Paraffin embedded tumour specimens were immunohistochemically stained for S-100, p53 and Ki67/MIB-1. FINDINGS: The most frequent initial symptoms were local swelling and pain followed by irritation of cranial nerves and spinal ataxia. Four tumours were localised at the head & neck region, three were found in the extremities and one tumour was located on the trunk. All patients underwent surgery with curative intent, but total resection, defined by negative surgical margins, was achieved in only 3 cases. All of these developed local recurrence with a mean disease free survival time of 10.6 months. In five cases, adjuvant radiation was given. During follow up, three patients developed distant metastases located in the lung, liver and subcutaneous tissue. Five out of eight patients died during follow-up with a mean survival time of 11.6 months after diagnosis. Results of immunohistochemical staining were as followed: S-100 (7/8), p53 (7/8). The Ki67/MIB-1 proliferation index was detectable in all tumour samples, it differed from 10-30%. INTERPRETATION: MPNST is a rare and fatal diagnosis in neurosurgery with high risk of local recurrence and occurence of distant metastases. Though mulitimodal therapy including surgical resection and adjuvant radiotherapy including brachytherapy is available, the prognosis remains dismal. Modern clinical studies and the development of effective chemotherapy is needed in order to gain control of the disease. PMID- 11437290 TI - Monitoring of facial evoked EMG for hemifacial spasm: a critical analysis of its prognostic value. AB - Microvascular decompression (MVD) has come to stay as an effective way of treating hemifacial spasm. But it remains to be seen how much each of the electrophysiological monitoring techniques (intra-operative) are contributing to its increased efficacy. Their role as indicators for re-exploration or recurrence is to be evaluated with more studies. We have used the lateral spread response in those patients who had distinctly abnormal recording on the ipsilateral side, studied the intra-operative changes during MVD and correlated with the outcome of surgery. 38 patients operated for HFS, were selected for intra-operative monitoring of abnormal muscle responses. In 17 patients, there was persistence of abnormal muscle responses in the immediate postoperative period and only 6 of them had mild HFS. Two of 21 patients who had disappearance of abnormal responses had persistent mild HFS; but in all cases, the HFS disappeared within 3 months. So we found that the intra-operative recording was really not reliable in predicting the immediate postoperative outcome. However the outcome at 3 months suggested that waiting for some time before re-exploration is a better option, especially if the HFS had become mild. PMID- 11437291 TI - The lateral transfacetal retrovascular approach for an anteriorly located chordoma originating from the second cervical vertebra. AB - BACKGROUND: Anteriorly located lesions at the craniocervical junction (CCJ) require careful surgical planning to avoid neuraxis retraction. Several different routes have been described in the search for the most appropriate yet least invasive approach. However, most of these far-lateral posterior approaches are specifically tailored for non-osseous intradural tumours or chordomas with cephalad extension. We introduce an approach that allows for better access to laterally extending osseous tumours originating from the second cervical vertebra. Using this approach, the lesion is manipulated from a strictly lateral direction parallel to a plane through the articular pillar of the CCJ and the odontoid process, and the C1- and C2-laminae are spared for posterior fixation. METHOD: The application of this approach is demonstrated in detail by an illustrative case of a chordoma originating from C2 that presented with intradural mass effect, considerable bone destruction, far-lateral extension to the right, and vertebral artery involvement. FINDINGS: The described approach gave ample access for total tumour resection. It allowed for safe control and displacement of the vertebral artery, spinal cord decompression, C2-corpectomy across the midline, and anterolateral bone reconstruction. No C1- or C2 hemilaminectomy was needed, and these bone elements could be used for posterior fixation (the patient presented in this study was referred to our institution after posterior fixation from the occiput to C3 had already been performed elsewhere). No intra-operative or postoperative complications occurred. At the 6 month follow-up, the patient was fully ambulatory with no neurological deficit. INTERPRETATION: The described lateral transfacetal route is the method of choice for operating on laterally extending osseous tumours originating from the second cervical vertebra. PMID- 11437292 TI - Lumbar disc surgery in regional anaesthesia--40 years of experience. AB - Lumbar disc operations have been performed in Brno since 1952 and from the very beginning they are performed under regional anaesthesia. Since 1965 until 1999 about 16,000 operations for herniated discs have been performed here. The number of operations doubled with the foundation of the second neurosurgical department in Brno in 1992. Lumbar disc surgery represents 25% of the operation spectrum in our department. Epidural anaesthesia is advantageous for less risk of some complications compared with general anaesthesia and enables communication between the surgeon and the patient. Other advantages are: less visible bleeding in the operative field due to the lateral position and the fact that the surgeon can sit during the operation. On a long-term basis we found 4-5% had further surgery for reccurent problems. Objective neurological evaluation 6 weeks after the operation showed improvement in 92% of patients. The patients themselves consider their status as very good or satisfactory in 82% (median 3 years after the operation). Besides a correctly performed operation we emphesize also the role of correct diagnosis (good correlation between the patient's problems and CT scan finding) for a good treatment result. PMID- 11437293 TI - Nitric oxide synthase inhibition attenuates vasoactive response to spinal cord stimulation in an experimental cerebral vasospasm model. AB - BACKGROUND: The basic mechanism of delayed cerebral vasospasm following subarachnoid haemorrhage (SAH) has been intensively investigated. It is thought that nitric oxide (NO) is a basic mediator of the cerebral vasodilator mechanism. Previous clinical and experimental studies have shown a cerebral vasodilator effect of high cervical spinal cord stimulation (SCS) however, the mechanism of this effect is still controversial. We investigated the contribution of the vasodilator effect of NO to this mechanism in an experimental SAH model using rabbits. METHOD: Four experimental groups, were designated: Group 1. Cerebral blood flow (CBF) was evaluated by transcranial Doppler ultrasonography (TDU) in 8 rabbits. Group 2. In 4 animals, intracisternal saline injection and cervical epidural electrode placement without SCS were performed before TDU. Group 3. TDU was performed before and after SCS on the fourth day of SAH in 8 rabbits. Group 4. In 8 animals, N-Nitro-L-Arginine Methyl Esther (L-NAME) was administered intracisternally on the fourth day of SAH, at a dose of 0.6 mg/kg, 45 minutes before SCS. CBF parameters, obtained via measurements or calculations from TDU data, were compared. FINDINGS: The occurrence of vasospasm after SAH was demonstrated with significant changes in TDU parameters (high peak systolic velocity and positive values of the degree of stenosis). In all SAH animals, SCS resulted in significant vasodilation. Even after the injection of L-NAME, SCS still had a significant vasodilatory effect in SAH animals, but there was also a significant difference in CBF parameters in the SCS-only group when compared with the L-NAME treatment before SCS group. INTERPRETATION: The mechanism of the cerebral vasodilatory effect of SCS remains controversial. Our results revealed the contribution of a neurohumoral effect which can be partially prevented by use of an NO synthase inhibitor. PMID- 11437294 TI - Specificity of reinnervation and motor recovery after interposition of an artificial barrier between transected and repaired nerves in adjacency--an experimental study in the rat. AB - Non-specific re-innervation of target organs caused by misdirected axonal growth at the repair site is regarded as one reason for a poor functional outcome after peripheral nerve transsection and repair. This study investigates the rate of aberrant re-innervation and its influence on motor recovery in the rat sciatic nerve using artificial sheets as barrier between tibial and peroneal nerves. The sciatic nerve was transsected and repaired as follows: epineural sutures (A x 6), fascicular repair of tibial and peroneal nerves respectively (B x 8), and the same as in group B, but separating both nerves using an Integra-sheet with silicone (C x 8), or Integra without silicone (D x 8). As control, solely the tibial nerve was transsected and repaired (E x 5). Final investigations after 4 months revealed that in group C, 50% of the Integra-silicone sheets were dislocated. No dislocation was found in group D. Muscle contraction force of the gastrocnemius muscle was significantly higher in group E as compared to all other groups. However although not significant, group D showed a consistently higher muscle contraction force than groups A, B, and C. Histology in groups A, B, and C with dislocated sheets demonstrated multiple axons growing from the tibial to the peroneal nerve and vice versa. In groups D and E, no such axonal growth was visible. These findings were confirmed by a significantly higher rate of specific reinnervation of the soleus muscle using sequential retrograde double labelling technique. Results of this study suggest that an artificial sheet such as Integra bears the potential of preventing aberrant re-innervation between repaired adjacent nerves resulting in improved motor recovery. Clinically, this technique may be of importance for brachial plexus, sciatic nerve, and facial nerve repair. PMID- 11437295 TI - Idiopathic spinal cord herniation: case report and literature review. AB - We report one case of spontaneous thoracic spinal cord herniation presenting with a progressive spastic paraparesis for 4 years in a 55 years old man. From preoperative MRI, showing a ventrally displaced atrophic spinal cord at T2-T3 level, a dorsal intradural arachnoid cyst was suspected. At operation, after a 3 level laminectomy, no arachnoid cyst was found and spinal cord herniation into a meningeal diverticulum was confirmed. The herniated myelon was replaced intradurally and the lumen of the diverticulum was filled with Teflon settled with fibrin glue to prevent recurrence. Postoperatively some neurological recovery was achieved. The literature was reviewed, regarding clinical and epidemiological features, proposed pathophysiological mechanisms, treatment options and outcome. Only 32 surgically proved cases of thoracic spinal cord herniation with no past history of spine trauma, injury or surgery were found. PMID- 11437296 TI - Recurrent neurohypophyseal germinoma causing invasion localized to temporal bone marrow-unreported neuroimaging studies compared to autopsy findings. PMID- 11437297 TI - Migration of pacemaker lead into the spinal venous plexus: case report with special reference to Batson's theory of spinal metastasis. AB - OBJECTIVE AND IMPORTANCE: Migration of a foreign material via venous routes into the spinal canal is a very rare incidence. We report the second case in which a foreign body has migrated into the spinal canal via the venous route. CLINICAL PRESENTATION: This 35-years-old man presented with sudden onset of severe low back pain and pain in the right leg four months after an unsuccessful attempt to remove a disconnected cardiac pacemaker lead via the femoral vein. Direct lumbar x-ray demonstrated the broken lead of the cardiac pacemaker at the entrance of the right L5 foramen which was also demonstrated by lumbar CT. SURGICAL INTERVENTION: After right L5 hemilaminotomy, the pacemaker lead was found in a vein of the anterior spinal venous plexus just beneath and lateral to the right L5 root. After dissecting it from the surrounding adipose tissue, the embolised pacemaker lead was taken out. CONCLUSION: We present a case report and review of the literature on migration of foreign material into the spinal canal, factors effecting the flow directions in the spinal veins. This case may be the first evidence that proved Batson's theory of spinal metastases in man. PMID- 11437298 TI - A large densely calcified lesion at the origin of both carotid arteries: brief report of a special case. PMID- 11437299 TI - Money, kisses, and electric shocks: on the affective psychology of risk. AB - Prospect theory's S-shaped weighting function is often said to reflect the psychophysics of chance. We propose an affective rather than psychophysical deconstruction of the weighting function resting on two assumptions. First, preferences depend on the affective reactions associated with potential outcomes of a risky choice. Second, even with monetary values controlled, some outcomes are relatively affect-rich and others relatively affect-poor. Although the psychophysical and affective approaches are complementary, the affective approach has one novel implication: Weighting functions will be more S-shaped for lotteries involving affect-rich than affect-poor outcomes. That is, people will be more sensitive to departures from impossibility and certainty but less sensitive to intermediate probability variations for affect-rich outcomes. We corroborated this prediction by observing probability-outcome interactions: An affect-poor prize was preferred over an affect-rich prize under certainty, but the direction of preference reversed under low probability. We suggest that the assumption of probability-outcome independence, adopted by both expected-utility and prospect theory, may hold across outcomes of different monetary values, but not different affective values. PMID- 11437300 TI - On the affine structure of perceptual space. AB - Affine geometry is a generalization of Euclidean geometry in which distance can be scaled along parallel directions, though relative distances in different directions may be incommensurable. This article presents a new procedure for testing the intrinsic affine structure of a psychological space by having subjects perform bisection judgments over multiple directions. If those judgments are internally consistent with one another, they must satisfy a theorem first proved by Pierre Varignon around 300 years ago. In the experiment reported here, this procedure was employed to measure the perceived structure of a visual ground surface. The results revealed that observers' judgments were systematically distorted relative to the physical environment, but that the judged bisections in different directions had an internally consistent affine structure. Implications of these findings for other possible response tasks are considered. PMID- 11437301 TI - A psychophysiological examination of cognitive processing of and affective responses to social expectancy violations. AB - Several models of person perception predict that expectancy violations have both affective and cognitive consequences for the perceiver. Although extant evidence generally supports these claims, the temporal resolution of traditional self report measures has limited researchers' ability to convincingly link underlying physiological processes with observed outcomes. In this study, we examined these issues by measuring brain (event-related brain potentials) and peripheral (facial electromyogram) electrophysiological activity while participants read positive and negative expectancy-consistent, expectancy-violating, expectancy-irrelevant, and semantically incongruent behavioral sentences about fictitious characters. The electromyogram results indicated that negative (but not positive) expectancy violating behaviors elicited enhanced negative affect as early as 100 to 300 ms poststimulus. The event-related potentials showed enhanced positivities with latency exceeding 300 ms in response to expectancy violations and negative behaviors. Semantically incongruent sentence endings influenced a separate negative component (N400), suggesting fundamental differences between semantic- and behavior-consistency processing. This difference also was evident in participants' recall. Implications for theoretical models of expectancy violation are discussed. PMID- 11437302 TI - Visual prior entry. AB - It has long been claimed that attended stimuli are perceived prior to unattended stimuli--doctrine of prior entry. Most, if not all, studies on which such claims have been based, however, are open to a nonattentional interpretation involving response bias, leading some researchers to assert that prior entry may not exist. Given this controversy, we introduce a novel methodology to minimize the effect of response bias by manipulating attention and response demands in orthogonal dimensions. Attention was oriented to the left or right (ie., spatially), but instead of reporting on the basis of location, observers reported the order (first or second) of vertical versus horizontal line segments. Although second order response biases were demonstrated, effects of attention in accordance with the law of prior entry were clearly obtained following both exogenous and endogenous attentional cuing. PMID- 11437303 TI - Perceptual load and visuocortical processing: event-related potentials reveal sensory-level selection. AB - Behavioral evidence suggests that the processing of parafoveal stimuli decreases as the perceptual demands of a task at fixation increase. However, it remains unclear whether or not this effect of perceptual load occurs during initial sensory-level processing at early stages of visuocortical analysis. Using event related potential measures, we found that increasing the perceptual load of foveal targets led to a significant decrease in the sensory-evoked response to parafoveal stimuli. Moreover, these effects were observed using two different operational definitions of perceptual load. This result indicates that perceptual load affects the flow of information during the initial stages of visuocortical processing. PMID- 11437304 TI - Visual search remains efficient when visual working memory is full. AB - Many theories of attention have proposed that visual working memory plays an important role in visual search tasks. The present study examined the involvement of visual working memory in search using a dual-task paradigm in which participants performed a visual search task while maintaining no, two, or four objects in visual working memory. The presence of a working memory load added a constant delay to the visual search reaction times, irrespective of the number of items in the visual search array. That is, there was no change in the slope of the function relating reaction time to the number of items in the search array, indicating that the search process itself was not slowed by the memory load. Moreover, the search task did not substantially impair the maintenance of information in visual working memory. These results suggest that visual search requires minimal visual working memory resources, a conclusion that is inconsistent with theories that propose a close link between attention and working memory. PMID- 11437305 TI - African Americans and high blood pressure: the role of stereotype threat. AB - We examined the effect of stereotype threat on blood pressure reactivity. Compared with European Americans, and African Americans under little or no stereotype threat, African Americans under stereotype threat exhibited larger increases in mean arterial blood pressure during an academic test, and performed more poorly on difficult test items. We discuss the significance of these findings for understanding the incidence of hypertension among African Americans. PMID- 11437306 TI - Walking while memorizing: age-related differences in compensatory behavior. AB - This study investigated predictions of the life-span theory of selection, optimization, and compensation, focusing on different patterns of task priority during dual-task performance in younger and older adults. Cognitive (memorizing) and sensorimotor (walking a narrow track) performance were measured singly, concurrently, and when task difficulty was manipulated. Use of external aids was measured to provide another index of task priority. Before dual-task testing, participants received extensive training with each component task and external aid. Age differences in dual-task costs were greater in memory performance than walking, suggesting that older adults prioritized walking over memory. Further, when given a choice of compensatory external aids to use, older adults optimized walking, whereas younger adults optimized memory performance. The results have broad implications for systemic theories of cognitive and sensorimotor aging, and the costs and benefits of assistive devices and environmental support for older populations. PMID- 11437307 TI - Numerical subtraction in the pigeon: evidence for a linear subjective number scale. AB - When humans and animals compare two numbers, responding is faster and more accurate with increasing numerical disparity and decreasing numerical size. Researchers explaining these distance and size effects often, assume that the subjective number continuum is logarithmically compressed. An alternative hypothesis is that the subjective number continuum is linear, but positions farther along it are proportionately fuzzier, that is, less precisely located. These two hypotheses have been treated as functionally equivalent because of their similar empirical predictions. The current experiment sought to resolve this issue with a paradigm originally developed to address the subjective representation of time (time left). In our adaptation, pigeons were required to compare a constant number with the number remaining after a numerical subtraction. Our results indicate that subjective number is linearly, not logarithmically, related to objective number. PMID- 11437308 TI - Subtracting pigeons: logarithmic or linear? PMID- 11437309 TI - Arousal, mood, and the Mozart effect. AB - The "Mozart effect" refers to claims that people perform better on tests of spatial abilities after listening to music composed by Mozart. We examined whether the Mozart effect is a consequence of between-condition differences in arousal and mood. Participants completed a test of spatial abilities after listening to music or sitting in silence. The music was a Mozart sonata (a pleasant and energetic piece) for some participants and an Albinoni adagio (a slow, sad piece) for others. We also measured enjoyment, arousal, and mood. Performance on tbe spatial task was better following the music than the silence condition but only for participants who heard Mozart. The two music selections also induced differential responding on the enjoyment, arousal and mood measures. Moreover, when such differences were held constant by statistical means, the Mozart effect disappeared. These findings provide compelling evidence that the Mozart effect is an artifact of arousal and mood. PMID- 11437310 TI - Not all laughs are alike: voiced but not unvoiced laughter readily elicits positive affect. AB - We tested whether listeners are differentially responsive to the presence or absence of voicing, a salient, distinguishing acoustic feature, in laughter. Each of 128 participants rated 50 voiced and 20 unvoiced laughs twice according to one of five different rating strategies. Results were highly consistent regardless of whether participants rated their own emotional responses, likely responses of other people, or one of three perceived attributes concerning the laughers, thus indicating that participants were experiencing similarly differentiated affective responses in all these cases. Specifically, voiced, songlike laughs were significantly more likely to elicit positive responses than were variants such as unvoiced grunts, pants, and snortlike sounds. Participants were also highly consistent in their relative dislike of these other sounds, especially those produced by females. Based on these results, we argue that laughers use the acoustic features of their vocalizations to shape listener affect. PMID- 11437311 TI - Parents explain more often to boys than to girls during shared scientific thinking. AB - Young children's everyday scientific thinking often occurs in the context of parent-child interactions. In a study of naturally occurring family conversation, parents were three times more likely to explain science to boys than to girls while using interactive science exhibits in a museum. This difference in explanation occurred despite the fact that parents were equally likely to talk to their male and female children about how to use the exhibits and about the evidence generated by the exhibits. The findings suggest that parents engaged in informal science activities with their children may be unintentionally contributing to a gender gap in children's scientific literacy well before children encounter formal science instruction in grade school. PMID- 11437312 TI - Emotional closeness as a mediator of the effect of genetic relatedness on altruism. AB - Inclusive fitness theory has been used to explain the overall pattern of altruistic behavior. However, this theory does not address the proximal causes of altruism. The purpose of the present study was to increase the understanding of altruism by testing a model that includes both ultimate and proximate causes. In particular, emotional closeness was tested as a mediator of the effect of genetic relatedness on altruistic behavior. This was accomplished by having college students choose which of their family members they would most likely provide with life-saving assistance. As expected, results showed that emotional closeness is an important proximal cause of altruism that partially mediates the effect of genetic relatedness on willingness to act altruistically. PMID- 11437313 TI - Mucosal immunization with Salmonella typhimurium expressing Lassa virus nucleocapsid protein cross-protects mice from lethal challenge with lymphocytic choriomeningitis virus. AB - OBJECTIVES: Lassa fever virus (LAS) is transmitted to man by rodent carriers and is fatal in a third of untreated cases. Our goal is to provide immune protection from Lassa fever by mucosal vaccination. STUDY DESIGN/METHODS: Mice were vaccinated intragastrically with control vectors or with vectors (vaccinia or Salmonella) expressing LAS nucleocapsid protein (NP). Mice were challenged intracranially with a lethal dose of the related arenavirus, lymphocytic choriomeningitis virus (LCMV), as a measure of the vaccine's ability to elicit cross-protection. RESULTS: Salmonella and vaccinia vectors expressing LAS NP each protected a third of the mice from lethal challenge with LCMV. All mice vaccinated with a vector expressing LCMV NP were protected as expected. CONCLUSIONS: The LAS recombinant Salmonella vector is comparable to the LAS recombinant vaccinia vector in its ability to cross-protect mice from lethal challenge. Nucleocapsid protein is an inadequate immunogen on its own, but provides sufficient cross-protection to make it a useful component of a broadly reactive arenavirus vaccine. PMID- 11437314 TI - Negative human herpesvirus 8 serology in sarcoidosis. PMID- 11437315 TI - Potent neutralization of primary human immunodeficiency virus clade C isolates with a synergistic combination of human monoclonal antibodies raised against clade B. AB - OBJECTIVES: We investigated the ability of several human neutralizing monoclonal antibodies (mAbs), originally raised against human immunodeficiency virus (HIV) clade B isolates, to neutralize primary clade C isolates as single agents and in combination. STUDY DESIGN/METHODS: HIV clade C isolates from five different countries were tested for susceptibility to neutralization by anti-clade B mAbs in human peripheral blood mononuclear cells. Monoclonal antibody combinations were evaluated for possible synergy. RESULTS: All 20 primary HIV clade C isolates could be neutralized 97.5% to 100% by a quadruple combination of mAbs IgG1b12, 2G12, 2F5, and 4E10. These mAbs recognized conserved epitopes and were highly synergistic, resulting in strong cross-clade neutralization. CONCLUSIONS: In our previous experiment, a synergistic combination of human neutralizing mAbs protected all macaque neonates against oral challenge with a simian-human immunodeficiency virus encoding HIV env. Together, our data suggest that passive immunization with currently available anti-clade B mAbs could play a role in preventing HIV clade C transmission through breastfeeding. PMID- 11437316 TI - Human herpesvirus 8 viral FLICE-inhibitory protein inhibits Fas-mediated apoptosis through binding and prevention of procaspase-8 maturation. AB - Viral FLICE-inhibitory proteins (v-FLIPs) encoded by several herpesviruses and poxviruses share the ability to inhibit apoptosis after engagement of death receptors. In the current article, we provide insights into the mechanisms by which the v-FLIP of human herpesvirus 8 (HHV-8) (also referred to as Kaposi's sarcoma-associated virus) protects cells from apoptosis after Fas-induced signaling. Using v-FLIP expression vectors, our results clearly show that HHV-8 v FLIP reduces the cleavage of procaspase-8 into its active p18 and p10 protease subunits upon Fas-induced cell death. These results were confirmed by lower caspase-8 and caspase-3 protease activities in extracts of HeLa cells expressing HHV-8 v-FLIP. Coimmunoprecipitation studies further indicate that HHV-8 v-FLIP physically interacts with procaspase-8, but not with Fas-associated protein with death domain in the cellular cytoplasm. These results suggest that binding of HHV 8 v-FLIP to procaspase-8 affects the recruitment and the activation of the latter at the death-induced signaling complex, resulting in diminished apoptotic cascade initiation. Because cellular FLIP was recently reported to modulate promoter containing NF-kappaB motifs and that both HHV-8 and human immunodeficiency virus type 1 (HWV-1) can infect monocytes, we studied the effects of v-FLIP on HIV-1 gene expression. Cotransfection experiments indicated that v-FLIP expression is associated with activation of HIV long terminal repeats: events that were strictly dependent on the presence of NF-kappaB consensus elements. In conclusion, HHV-8 v-FLIP can possibly contribute to the pathogenesis of both HHV 8 and HIV-1 through impaired Fas-dependent killing of infected cells by cytotoxic T cells and through activation of HIV gene expression. PMID- 11437317 TI - Cap and polyA tail enhance translation initiation at the hepatitis C virus internal ribosome entry site by a discontinuous scanning, or shunting, mechanism. AB - OBJECTIVES: Hepatitis C virus (HCV) does not replicate in vitro, suggesting that cultured cells may lack factors that are essential for efficient use of HCV messenger RNAs (mRNAs). Here, we have studied the efficiency of HCV mRNA translation compared with translation of capped and polyadenylated mRNAs in human cells. STUDY DESIGN/METHODS: We have generated noninfectious minivirus mRNAs from an infectious HCV genome. These mRNAs were transfected into human cells, and the translation efficiency was determined. RESULTS: Hepatitis C virus mRNAs under control of the HCV internal ribosome entry site (IRES) were inefficiently translated compared with capped and polyadenylated mRNAs. Addition of a cap and a polyA tail on the HCV mRNAs revealed that these structures interacted with the hepatitis C IRES in a synergistic manner to load ribosomes onto the HCV mRNAs, thereby strongly enhancing translation. The positive effect of the cap and the polyA tail on initiation of translation at the initiator AUG embedded in the HCV IRES was the result of a discontinuous scanning, or shunting, mechanism. CONCLUSIONS: The results demonstrated that recruitment of ribosomes to the HCV mRNAs was inefficient in dividing cultured cells. Factors that are necessary for efficient translation of the HCV mRNAs in hepatocytes may be absent or inactive in cultured cells. This may be one reason for the inefficient replication of the HCV in vitro. PMID- 11437318 TI - Human papillomavirus-16-E7 oncoprotein enhances the expression of adhesion molecules in cervical endothelial cells but not in human umbilical vein endothelial cells. AB - OBJECTIVES: E7 is one of the oncoproteins encoded by human papillomavirus-16 (HPV 16), the major etiologic factor responsible for cervical cancer. Human papillomavirus-16-E7 expressed by human uterine cervix carcinoma cells is also released in the extracellular compartment where it induces immune suppression. We investigated whether E7 was also responsible for the enhanced endothelial adhesiveness required in cancer progression. STUDY DESIGN/METHODS: We treated cervical microvascular endothelial cells (CrMVEn) and human umbilical vein endothelial cells (HUVEC) with E7, tumor necrosis factor-alpha (TNF-alpha), and hydrogen peroxide (H2O2) and measured the expression of E-selectin, intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) by fluorescent-activated cell sorter analysis. RESULTS: E7 strongly induced the expression of E-selectin, ICAM-1, and VCAM-1 in CrMVEn, but not in HUVEC. Tumor necrosis factor-alpha further increased the endothelial expression of adhesion molecules in CrMVEn. Hydrogen peroxide pre-treatment resulted in an enhanced ICAM 1 and a decreased E-selectin and VCAM-1 expression. We also show indirect effects when endothelial cells were stimulated with the supernatant of E7-pretreated macrophages. CONCLUSIONS: These results show that HPV-16-E7 oncoprotein strongly induces adhesion molecules expression in organ-specific endothelial cells. PMID- 11437319 TI - Expression of human herpesvirus 8 (HHV-8)-encoded immediate early protein, open reading frame 50, in HHV-8-associated diseases. AB - OBJECTIVES: To investigate the expression of the open reading frame (ORF) 50 protein, a human herpesvirus 8 (HHV-8)-encoded immediate early protein, in HHV-8 associated diseases. STUDY DESIGN/METHODS: We developed a rabbit anti-ORF50 protein polyclonal antibody, and investigated the ORF50 protein expression by immunohistochemistry using primary effusion lymphoma (PEL) cell lines and tissue sections of Kaposi's sarcoma (KS), HHV-8-associated solid lymphoma, and multicentric Castleman's disease (MCD). RESULTS: Western blot analysis revealed that this antibody reacted with a 110-kd protein in the lysate of phorbolester stimulated HHV-8-infected PEL cell lines. Immunohistochemistry revealed that a very small population of cells expressed the ORF50 protein in KS and HHV-8 associated solid lymphoma cells, and almost all these cells expressed HHV-8 encoded latency-associated nuclear antigen. The ORF50 protein expression was also rare in the cells of PEL cell lines, and the staining pattern was diffuse or dot like in the nuclei, overlapping partially with that of promyelocytic leukemia protein. In MCD, however, the ORF50 protein was expressed in some cells of the mantle zone of the lymphoid follicle. CONCLUSIONS: The ORF50 protein expression in vivo was considerably rare in KS, HHV-8-associated solid lymphoma, and PEL, but was more frequent in MCD. Rare expression of this transactivator protein in HHV-8-associated malignancies causes low expression levels of other lytic proteins and may play a role in the maintenance of the latent infection. This is the first report describing the expression of an immediate early protein of HHV-8 in cases of HHV-8-associated diseases. PMID- 11437320 TI - Resistance of hepatitis B virus to antiviral drugs: current aspects and directions for future investigation. AB - Despite the existence of vaccines, chronic hepatitis B virus (HBV) infection remains a major health problem worldwide. Interferon therapy successfully controls infection in only a small percentage of chronically infected individuals. The recent approval of the nucleoside analogue lamivudine for the treatment of chronic HBV infection has ushered in a new era of antiviral therapy. While lamivudine is highly effective at controlling viral infection short-term, prolonged therapy has been associated with an increasing incidence of viral resistance. Thus, it appears that lamivudine alone will not be sufficient to control chronic viral infection in the majority of individuals. In addition to lamivudine, several new nucleoside and nucleotide analogues that show promising antihepadnaviral activity are in various stages of development. Lamivudine resistance has been found to confer cross-resistance to some of these compounds and it is likely that resistance to newer antivirals may also develop during prolonged use. Drug resistance therefore poses a major threat to nucleoside analogue-based therapies for chronic HBV infection. Fortunately, combination chemotherapy (antiviral therapy with two or more agents) can minimize the chance that resistance will develop and can be expected to achieve sustained reductions in viral load, provided that suitable combinations of agents are chosen. Here we review the basis of drug resistance in HBV, with emphasis on aspects that are likely to affect drug choice in future. PMID- 11437321 TI - Structure-activity relationship studies on new DABOS: effect of substitutions at pyrimidine C-5 and C-6 positions on anti-HIV-1 activity. AB - Several 5-alkyl, 5-alkenyl, 5-iso-alkyl, 5-halo, 5-aminomethyl and 5-carboxy derivatives of S-DABOs (dihydro-alkyl (or cyclo-alkyl)thio-benzyloxopyrimidines), DATNOs (dihydro-alkylthionaphthylmethyl-oxopyrimidines) and F2-S-DABOs (dihydro alkyl (or cyclo-alkyl)thio-2,6-difluorobenzyl-oxopyrimidines) have been prepared and tested as anti-HIV-1 agents. S-DABO derivatives bearing at C-6 position monosubstituted phenylmethyl or heteroarylmethyl units have also been synthesized. 2-Alkylthio-3,4-dihydropyrimidin-4(3H)-one derivatives of F2-S-DABO series bearing small alkyl groups at C-5 proved to be potent inhibitors of HIV-1 replication in vitro with selectivity indexes ranging from 250 to >2,500. PMID- 11437322 TI - Characterization of inhibitory action of concanamycins against herpes simplex virus. AB - Concanamycins A (Conmy A) and B (Conmy B), well-known inhibitors of the vacuolar proton-ATPase, were isolated from the culture broth of Streptomyces sp. strain FK51 as antiherpetic agents. These compounds showed potent inhibition of herpes simplex virus type 1 (HSV-1) replication in an in vitro assay system, having antiviral activities with 50% inhibitory concentrations of 0.072 and 0.51 ng/ml for Conmy A and Conmy B, respectively. While the attachment of HSV-1 to Vero cells was not inhibited, both of the compounds blocked the penetration of virus into host cells. When added to the late stages of virus replication, the concanamycins also exerted marked inhibitory effects on the production of viruses. Release of progeny viruses was found to be suppressed by the agents. SDS PAGE analysis of isotope-labelled HSV-specific proteins revealed that the synthesis of beta proteins was moderately inhibited and some of the glycoproteins were synthesized with reduced molecular weights. Western blot analysis using antibodies against two HSV-specific glycoproteins (gC and gD) showed differences in their syntheses between untreated and Conmy A-treated cells. Syncytium formation by HSV-1 strain HF was inhibited, and small plaques with rounded cells were formed in Conmy A-treated cell cultures. When wild-type HSV-1 was serially propagated under the selective pressure of Conmy A, and the resulting progeny viruses were grown in drug-free medium, their plaque morphology of syncytium and sensitivity to Conmy A were the same as those of parent virus. From these findings, antiherpetic activities of Conmy A and B might be mainly dependent on their activities as vacuolar proton-ATPase inhibitors with intracellular translocation of glycoproteins and the inhibition of the maturation of virus glycoproteins. PMID- 11437323 TI - In vitro and in vivo activity of 1-O-hexadecylpropanediol-3-phospho-ganciclovir and 1-O-hexadecylpropanediol-3-phospho-penciclovir in cytomegalovirus and herpes simplex virus infections. AB - Human cytomegalovirus (HCMV) and herpes simplex virus (HSV) can cause a wide variety of clinical manifestations in man. Ganciclovir (GCV) is effective against HCMV infection when administered by the intravenous route and may be used orally in large doses for prophylaxis of HCMV infections in organ transplantation patients and in AIDS patients. In previous studies with acyclovir (ACV), we found that covalent attachment of an alkyl glycerol phosphate moiety greatly increased oral bioavailability and increased antiviral activity against hepatitis B virus. Adducts of ACV with alkyl propanediol phosphate were more active than the alkyl glycerol phosphate analogue in vitro in 2.2.15 cells, which constitutively produce hepatitis B virus. To see if this strategy would work for two other poorly absorbed nucleoside analogues, we synthesized 1-O-hexadecylpropanediol-3 phospho-GCV (HDP-P-GCV) and 1-O-hexadecyl-propanediol-3-phospho-penciclovir (HDP P-PCV), and evaluated the in vitro antiviral activity, selectivity and oral antiviral activity of both compounds versus GCV or PCV in mice infected with HSV 1 or HDP-P-GCV versus murine cytomegalovirus (MCMV). HDP-P-GCV is orally active in both MCMV and HSV-1 infection in mice with antiviral activity equivalent to (HSV-1) or greater than oral GCV (MCMV). Oral HDP-P-PCV was more active than PCV orally versus intranasal HSV-1 infection in mice. PMID- 11437324 TI - Treatment of lethal vaccinia virus respiratory infections in mice with cidofovir. AB - Intranasal infection of BALB/c mice with the WR strain of vaccinia virus leads to pneumonia, profound weight loss and death. Five days after intranasal inoculation, virus from untreated mice was recovered from 11 organs, tissues and whole blood. The highest titres [>10(8) plaque forming units (pfu)/g] were in lungs and nose/sinus tissue, with about 10(7) pfu/g in spleen and blood. Seven other organs contained 30- to > or = 50-fold lower amounts of virus. Mice infected with the related cowpox virus (for comparative purposes) had the majority of virus located in the respiratory tract. The vaccinia mouse model was used to study the efficacy of cidofovir treatments on the infection. Subcutaneous injections of 30 or 100 mg/kg/day, given on days 1 and 4 after virus challenge, reduced mortality by 60-100%. However, lung virus titres on days 2-5 were reduced no more than 10-fold by these treatments. A moderate improvement in drug efficacy occurred with daily treatments for 5 days. The efficacy of cidofovir also increased as the virus challenge dose decreased, where subcutaneous or intraperitoneal treatment routes showed similar degrees of protection. Although it has been known for many years that the WR strain of vaccinia virus can cause lethal infections by intranasal route, its application to antiviral therapy represents a new model for studying anti-orthopoxvirus agents. PMID- 11437325 TI - Gene therapy of severe combined immunodeficiencies. AB - Recent advances in gene transfer in human hematopoietic cells, combined with a better understanding of the genetic aspects of several immunodeficiencies, has offered new opportunities in the domain of gene therapy. Severe combined immunodeficiency (SCID) appear to represent a good model for the application of gene therapy, combining an expected selective advantage for transduced cells, an absence of immunological response to the vector and/or the therapeutic transgene, together with accessibility to hematopoietic stem cells (HSC). Ex vivo retroviral transduction of a therapeutic transgene in HSC prior to transplantation appears to be a particularly effective and long-lasting means of restoring the expression of a mutated gene in the lymphoid lineage. Furthermore, encouraging therapeutic benefits as a result of a gene therapy protocol for the treatment of X-linked severe combined immunodeficiencies (SCID-X1) invites many questions as to the reasons for this therapeutic benefit. This review outlines the results that have been achieved in gene therapy for SCID-X1, ADA-SCID as well as other types of SCID, and discusses the possible relationship between the physiopathology of each disease and the success of relevant trials. PMID- 11437326 TI - Retroviral transduction of human CD34+ cells on fibronectin fragment CH-296 is inhibited by high concentrations of vector containing medium. AB - BACKGROUND: The objective of the present study was to optimize conditions for retroviral transduction of human cord blood (CB) CD34+ cells and to reveal mechanisms which interfere with efficient gene transfer. METHODS: An MSCV based retroviral vector with the gene for enhanced green fluorescent protein (MGIN) produced by GP+envAM12 (amphotropic envelope), PG13 (gibbon ape leukemia virus envelope) or 293GPG (vesicular stomatitis virus envelope) cell lines was used to transduce cord blood CD34+ cells on Retronectin (fibronectin fragment CH-296) in three different ways: either in vector containing medium (VCM), in fresh medium on Retronectin pre-loaded with vector or in VCM on Retronectin pre-loaded with vector. RESULTS: Paradoxically, the transduction efficiency obtained with pre load of vector onto Retronectin alone was higher than pre-load plus VCM for PG13 MGIN (67.9 +/- 6.0% vs 24.9 +/- 8.0%) and AM12-MGIN (47.5 +/- 5.8% vs 38.7 +/- 2.2%). Further experiments showed that transduction on Retronectin pre-loaded with PG13-MGIN or AM12-MGIN was inhibited by the presence of the same VCM at high concentrations, but not by the presence of a VCM with a different receptor specificity. If no pre-load of vector was performed, the highest transduction efficiencies were seen when VCMs were diluted 1:10 (MOIs of 3). The inhibitory effect of high titer PG13-MGIN VCM was confirmed in more primitive CD34+CD38low cells and in NOD/SCID repopulating cells, and was also seen in experiments with bone marrow CD34+ cells. CONCLUSIONS: Retroviral transduction of CB CD34+ cells on Retronectin is inhibited by high titer PG13 and GP+envAM12 vector containing medium. Efficient gene transfer to human hematopoietic cells can be obtained by preload alone of the vector onto Retronectin. These findings are of importance for the design of transduction protocols for repopulating hematopoietic cells. PMID- 11437327 TI - Optimisation of retroviral supernatant production conditions for the genetic modification of human CD34+ cells. AB - BACKGROUND: Clinically applicable protocols for ex vivo modification of human CD34+ hematopoietic stem/progenitor cells rely on incubation of the target cell with supernatant containing recombinant retroviral particles. Although components of the supernatant may have a profound impact on both preclinical and clinical outcome, to date supernatant production has not been properly addressed with regard to CD34+ cells. We wanted to investigate and optimise production conditions for this target using simple, reproducible and clinically applicable procedures and reagents. METHODS: Retroviral supernatant was obtained from producer cell GP+Am12 under various production conditions and tested for bulk transduction efficiency and endpoint titre on murine and human cell lines. Gene transfer efficiency into CD34+ cells from mobilised peripheral blood, after a single exposure to retroviral supernatant, was measured by transgene expression, colony forming assay and long-term culture colony forming assay. RESULTS: Bulk gene transfer or endpoint titre values obtained on cell lines for the different production conditions were not predictive of gene transfer efficiency into hematopoietic progenitors. Time of virus production appeared to have the greatest impact on gene transfer, peaking at 6 h and decreasing 2-3-fold at longer time points. Neither the culture vessel used nor the temperature for virus production had any significant effect on gene transfer into CD34+ cells. Supernatant could be produced under defined serum-free conditions as efficiently as serum containing conditions for CD34+ cell gene transfer. CONCLUSIONS: The present data provide important implications for the establishment of quality controls for small- and large-scale clinical grade supernatant production for gene transfer into human hematopoietic stem/progenitor cells. PMID- 11437328 TI - Adenovirus-mediated gene transfer in canine eyes: a preclinical study for gene therapy of human uveal melanoma. AB - BACKGROUND: Melanomas of the uveal tract are the most common intraocular malignancies in adults, with an incidence of six cases per million adults per year. Enucleation, which may enhance the dissemination of tumour cells into the systemic circulation, is still required for eyes with large tumours. Gene therapy is proposed as a new therapeutic approach for uveal melanoma management. METHODS: The potential of adenovirus-mediated gene transfer to normal eyes of two laboratory Beagles and in an iris tumour of a Great Dane were evaluated. Replication-defective adenoviral vectors (Adbetagal) were used to assess the feasibility, efficiency and safety of direct adenoviral delivery to the anterior chamber of normal eyes and to an iris tumour. The expression of angiostatin into the aqueous humour following an adenoviral-mediated delivery of human angiostatin (AdK3) was also investigated. RESULTS: The ciliary body was the area preferentially transduced after adenoviral injection into the anterior chamber. It was also demonstrated that a direct intratumoral injection of a recombinant adenovirus efficiently transduces a canine uveal melanoma. Western blot analysis performed on the aqueous humour revealed that the expression of the angiostatin recombinant protein in the aqueous humour correlated with the dose of AdK3 administered. Lymphocyte infiltrates at the site of AdK3 injection indicated induction of a strong cellular immune response, and humoral immune responses developed in all three dogs. CONCLUSIONS: The present study involving adenovirus mediated gene transfer to dog eyes provides an essential basis for gene therapy treatment of uveal melanoma-bearing patients. PMID- 11437329 TI - Engineered human mesenchymal stem cells: a novel platform for skeletal cell mediated gene therapy. AB - BACKGROUND: Human mesenchymal stem cells (hMSCs) are pluripotent cells that can differentiate to various mesenchymal cell types. Recently, a method to isolate hMSCs from bone marrow and expand them in culture was described. Here we report on the use of hMSCs as a platform for gene therapy aimed at bone lesions. METHODS: Bone marrow derived hMSCs were expanded in culture and infected with recombinant adenoviral vector encoding the osteogenic factor, human BMP-2. The osteogenic potential of genetically engineered hMSCs was assessed in vitro and in vivo. RESULTS: Genetically engineered hMSCs displayed enhanced proliferation and osteogenic differentiation in culture. In vivo, transplanted genetically engineered hMSCs were able to engraft and form bone and cartilage in ectopic sites, and regenerate bone defects (non-union fractures) in mice radius bone. Importantly, the same results were obtained with hMSCs isolated from a patient suffering from osteoporosis. CONCLUSIONS: hMSCs represent a novel platform for skeletal gene therapy and the present results suggest that they can be genetically engineered to express desired therapeutic proteins inducing specific differentiation pathways. Moreover, hMSCs obtained from osteoporotic patients can restore their osteogenic activity following human BMP-2 gene transduction, an important finding in the future planning of gene therapy treatment for osteoporosis. PMID- 11437330 TI - Strain-dependent anterior segment neovascularization following intravitreal gene transfer of basic fibroblast growth factor (bFGF). AB - BACKGROUND: A promising strategy for delaying death of photoreceptor cells in retinal degenerative disease is to support survival of these cells through intraocular delivery of growth/neurotrophic factors. One factor that has received great attention is basic fibroblast growth factor (bFGF; fgf-2), a known stimulator of angiogenesis. We evaluated the potential for neovascularization induced by adenovirus-mediated intravitreal delivery of bFGF. METHODS: Recombinant adenoviruses carrying the low molecular weight (18 kD) or the high molecular weight (22, 23 and 24 kD) forms of human bFGF, driven by the cytomegalovirus (CMV) promoter/enhancer, were prepared. Viruses were delivered to eyes of different strains of mice and rats through intravitreal injection. Contralateral eyes were injected with control virus carrying a reporter gene [green fluorescent protein (GFP) or lacZ]. Transgene expression was assessed by Western analysis and by immunohistochemistry. Neovascularization was evaluated in vivo and histologically at termination of the experiment. RESULTS: Adenovirus mediated delivery of the 18 kD form of bFGF resulted in anterior segment neovascularization in a strain-dependent fashion. Generation of new blood vessels was not observed after injection of the higher molecular weight forms of bFGF or of control solutions. CONCLUSION: The low molecular weight form (18 kD) (but not the high molecular weight forms) of bFGF drives angiogenic response in the anterior segment of specific strains of mice. Genetic modifiers may contribute to and/or prevent neovascularization induced by bFGF. PMID- 11437331 TI - Factors influencing cross-presentation of non-self antigens expressed from recombinant adeno-associated virus vectors. AB - BACKGROUND: We have previously demonstrated that recombinant adeno-associated virus vectors expressing the influenza virus hemagglutinin (rAAV-HA) in skeletal muscle results in T-cell priming and muscle fiber destruction due to cross presentation of HA by dendritic cells (DC). Based on controversial observations concerning the stability of non-self proteins expressed from rAAV vectors it is important to understand the factors influencing cross-presentation of transgene products following rAAV mediated gene transfer, in order to be able to use this vector safely in the clinic. METHODS: In order to understand the factors influencing in vivo cross-presentation of non-self proteins, we have retargeted the immunogenic lacZ protein in the context of rAAV from the cytoplasm to the cell surface and studied the activation of LacZ specific immune responses following intramuscular mediated gene transfer. In addition, using tools available for studying in vitro HA-specific T-cell activation, our aim was to identify the cell types involved in class I and class II restricted cross presentation as well as the nature of the cross-presented material. RESULTS: By retargeting the lacZ protein in the context of rAAV to the cell membrane, we found that one of the factors influencing the efficiency of cross-presentation of non-self antigens is the localization of the transgene product within the target cells. Following rAAV-LacZ mediated gene transfer to the muscle we demonstrated that the membrane-bound form of LacZ resulted in target cell destruction, which is in stark contrast to the stability observed with rAAV-LacZ vectors expressing the cytoplasmic form of LacZ. Using an in vitro assay, we were able to show that dendritic cells (DC) in addition to B-cells cross-presented HA to class II restricted T-cells whereas only the former were able to activate class I restricted CD8+ T-cells. High-dose antigens were needed for efficient class I restricted T-cell priming, whereas class II restricted T-cells were activated by less antigen. CONCLUSION: The present results indicate that immune responses to non-self antigens expressed from rAAV vectors depend on the accessibility of such antigens to different local antigen-presenting cells. PMID- 11437332 TI - A small, synthetic peptide for gene delivery via the serpin-enzyme complex receptor. AB - BACKGROUND: The serpin-enzyme complex receptor (SECR) has previously been successfully targeted for gene delivery using synthetic peptide ligands covalently linked in fluid phase to commercially available polylysine preparations (approximately 10-54kDa). The objective of the present study was to improve this approach by the use of small, bifunctional, and easily standardised synthetic peptides. METHODS: Two synthetic peptides designated polylysine antitrypsin 1 (PAT1) (K16 FNKPFVFLI) and PAT2 (K16 CSIPPEVKFNKPFVFLI) were evaluated for gene delivery to the HUH7 human hepatocyte cell line. The K16 moiety binds DNA electrostatically, while the FVFLM motif of human alpha1 antitrypsin targets the SECR. RESULTS: Both PAT1 and PAT2 bind to and condense DNA into small particles as shown by laser scattering techniques. However, only PAT2 is effective for gene delivery, presumably on account of the greater distance between the K16 chain and the FVFLM motif. Gene delivery by PAT2/DNA complexes is chloroquine-dependent, can be blocked completely by free ligand (CSIPPEVKFNKPFVFLI), and is highly efficient (e.g. approximately five-fold more effective than lipofectamine). At physiological salt concentrations, PAT2/DNA complexes formed at 4 microg/ml DNA are approximately 350 nm in diameter and highly effective for gene transfer, but at 100 microg/ml DNA the complexes are aggregated (diameter > 4 microm) and inactive. CONCLUSIONS: A small (33 amino acid), bifunctional, synthetic peptide represents a highly efficient and readily standardised DNA vector for the SECR. The effectiveness of this peptide depends on the distance of the K16 moiety from the targeting ligand. High salt concentrations are not required to form effective vector/DNA complexes. PMID- 11437333 TI - Time course of gene expression after plasmid DNA gene transfer to the liver. AB - BACKGROUND: High levels of expression in hepatocytes can be achieved after intraportal delivery of plasmid DNA vectors with up to 10% of all liver cells transfected. CMV promoter-driven expression is very high on Day 1 after injection, but is diminished strongly by Day 2. Expression slowly declines after 1 week. We describe experiments aimed at elucidating the reasons for this rapid decline in transgene expression. METHODS: Histological methods were used to determine the presence and extent of liver damage and hepatocyte proliferation. Viral and liver-specific promoters were tested to study promoter shut-off, Southern blotting was performed to determine the loss of the pDNA vector over time, and several mouse models were used to study the host immunological response. RESULTS: pDNA is lost rapidly early after injection, but remains at a relatively stable copy number after Day 4. Southern blotting experiments showed that plasmid DNA could be detected for at least 12 weeks after injection (0.2 copies per genome). The early rapid decline of expression is promoter dependent. A liver-specific albumin promoter resulted in similar levels of expression on Days 1 and 7, suggesting that promoter inactivation may be responsible for the instability of CMV promoter-driven expression. The slow decline in expression levels after 1 week appears to be the result of an immune response directed against the expressed transgene. Expression was much prolonged in immunosuppressed, immunodeficient, or antigen-tolerized mice. CONCLUSION: The present data suggest that if promoter inactivation can be overcome, intravascular delivery of plasmid DNA could be a highly efficient, simple and non-toxic liver gene therapy approach. Intravascular delivery of pDNA allows for the rapid screening of novel expression vectors in vivo. PMID- 11437334 TI - Regulation of transcription in Helicobacter pylori: simple systems or complex circuits? AB - A common strategy used by both Gram-negative and Gram-positive bacterial pathogens is based on the synchronisation of virulence gene expression using a variety of regulatory systems and networks to overcome host defence. During the last decade an exponentially growing number of studies on Helicobacter pylori, a human pathogen associated with diverse stomach diseases, have mainly focussed on the elucidation of mechanisms and functions of virulence factors. A subset of these studies were focussed on the molecular mechanisms regulating gene transcription in H. pylori with the aim of understanding the profound physiological changes that this pathogen, as well as other bacteria, undergoes during infection. Despite the limited number of putative regulatory proteins, as deduced from genome sequence analyses, evidence is accumulating for the existence of new and complex circuits regulating gene transcription and virulence of this bacterium. Here we will focus on the molecular mechanisms used by H. pylori to control gene transcription. PMID- 11437335 TI - The BvgAS two-component system of Bordetella spp.: a versatile modulator of virulence gene expression. AB - Bordetella pertussis and the closely related species B. parapertussis and B. bronchiseptica colonize the respiratory tract and cause related diseases in man or mammalian species, respectively. Expression of virulence factors by these pathogens is coordinately regulated by the BvgAS two-component system according to changes in the growth conditions. Signal transduction by the BvgAS system is characterized by a complex His-Asp-His-Asp phosphorelay. This system controls the expression of two distinct subsets of genes either in a positive (vag genes) or in a negative (vrg genes) manner. Most of the known virulence factors such as several toxins and adhesins are encoded by vag genes, whereas the functions of most vrg genes and the biological significance of the vrg regulon are not yet clear. This review discusses the current knowledge about the molecular mechanisms of virulence regulation and their relevance for infection by these respiratory pathogens. PMID- 11437336 TI - Quorum sensing and the regulation of virulence gene expression in pathogenic bacteria. AB - For many pathogens, the outcome of the interaction between host and bacterium is strongly affected by the bacterial population size. Coupling the production of virulence factors with cell population density ensures that the mammalian host lacks sufficient time to mount an effective defence against consolidated attack. Such a strategy depends on the ability of an individual bacterial cell to sense other members of the same species and in response, differentially express specific sets of genes. Such cell-cell communication is called "quorum sensing" and involves the direct or indirect activation of a response regulator by a small diffusible signal molecule. A number of chemically distinct quorum-sensing signal molecules have been described including the N-acyl-L-homoserine lactones (AHLs) in Gram-negative bacteria and post-translationally modified peptides in Gram positive bacteria. For example, the human pathogens Pseudomonas aeruginosa and Staphylococcus aureus employ AHLs and peptides, respectively, to control the expression of multiple virulence genes in concert with cell population density. Apart from their role in signal transduction, certain quorum-sensing signal molecules, notably N-(3-oxododecanoyl)homoserine lactone, possess intrinsic pharmacological and immunomodulatory activities such that they may function as virulence determinants per se. While quorum-sensing signal molecules have been detected in tissues in experimental animal model and human infections, the mutation of genes involved in either quorum-sensing signal generation or signal transduction frequently results in the attenuation of virulence. Thus, interference with quorum sensing represents a promising strategy for the therapeutic or prophylactic control of infection. PMID- 11437337 TI - Regulation of virulence genes in Listeria. AB - As in all pathogenic bacteria, virulence of the facultative intracellular Listeria species is a multifactorial trait. The expression of the bacterial genes involved in the different steps of the infectious process--invasion, intracellular multiplication and spreading--is temporally and spatially controlled, thus ensuring the presence of the respective gene products at the right moment and place. So far, one network which is involved in the regulation of listerial virulence, the PrfA regulon, has been characterized rather well. The key element of this regulon, PrfA, belongs to the Crp/Fnr family of transcriptional regulators. Its synthesis and activity are influenced by a variety of physico-chemical signals outside and inside of eukaryotic host cells. The analysis of virulence gene expression in vivo, i.e. in infected host cells, indicates that yet uncharacterized bacterial factors other than PrfA, and possibly also host factors, modulate the expression of the PrfA regulon. Essentially nothing is known about the signal transduction pathways involved in the observed differential expression of virulence genes. Fermentable carbon sources seem to have a particular role in virulence gene regulation. In addition to the PrfA regulon, the Clp stress proteins have an impact on Listeria virulence. These two regulons interact with each other by an unknown mechanism. PMID- 11437338 TI - Regulation of virulence determinants in Staphylococcus aureus. AB - The pathogenicity of Staphylococcus aureus depends on the combined action of more than 40 different extracellular toxins, enzymes and cell surface proteins. A global regulator agr controls the production of many of these virulence factors by a regulating RNA molecule, RNAIII. Most of the virulence genes regulated by RNAIII are also regulated by SarA and a family of homologous proteins. The Sar proteins appear to repress transcription of individual virulence genes or sets of genes. As some Sar proteins also repress one or more sar homologous genes an increased production of a single Sar protein can result in decreased expression of some virulence genes, and an increased expression of others. Results are presented suggesting that RNAIII might function as an antirepressor, binding one or more of the Sar proteins. PMID- 11437339 TI - Regulation of virulence genes in Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis has demonstrated remarkable ability to survive in diverse conditions encountered during the infection process. These involve surviving the bactericidal stresses within the macrophage, the anaerobic and nutritionally altered environment of the granuloma, and the metabolically inactive latent state. Understanding the molecular basis of this adaptive behavior lies in the identification of genes (or virulence determinants) specifically expressed under these varied conditions. Transcriptional control plays a key role in regulating gene expression in response to environmental signals. However, even after decades of investigation our knowledge about the function of these regulatory mechanisms in mycobacteria remains meagre. But the elucidation of the genome sequence and implementation of sophisticated molecular genetic approaches to this organism have made a revolutionary impact on the study of mycobacterial pathogenesis. Deletion and complementation of individual genes can be done at will facilitating the comparative analysis of mutants and wild type strains. Novel and powerful technologies such as DNA microarrays, fluorescent beacons and proteomics have made possible the analysis of the expression levels of multiple genes in in vitro systems. More technically challenging uses of these techniques is being undertaken to explore pathogen gene expression within the host. This will lead to the identification of virulence factors and give definitive insight into their regulatory signals. PMID- 11437340 TI - Gene regulation and host adaptation mechanisms in Candida albicans. AB - The yeast Candida albicans is a harmless member of the normal microflora on the mucosal surfaces of most healthy persons, but it can cause severe opportunistic infections in immunosuppressed patients. To become a successful human commensal and pathogen, C. albicans has evolved host adaptation mechanisms on different levels. The regulated expression of virulence and other genes in response to environmental signals allows an optimal adaptation to new host niches during the course of an infection. In addition, C. albicans is able to switch between different cell types in a reversible and apparently random fashion. Phenotypic switching involves the coordinated regulation of phase-specific genes, and the resulting generation of selected, pre-programmed cell types may represent an additional strategy to adapt to certain host environments. Finally, C. albicans produces genetically altered variants at a high rate. This microevolution ensures survival when the pathogen encounters new adverse conditions, as exemplified by the development of stable drug-resistant variants under the selection pressure caused by antimycotic therapy. Thus, rather than the possession of single dominant virulence factors, it is its remarkable versatility that makes C. albicans the most important fungal pathogen of humans. PMID- 11437341 TI - Iron uptake mechanisms and their regulation in pathogenic bacteria. AB - In the human body iron is present in growth-limiting amounts for bacteria. For this reason intricate iron transport and iron regulatory systems evolved in bacteria to guarantee a sufficient iron supply. The few principal mechanisms that underly the large variety of iron supply systems will be presented, as well as cases, in which defined iron supply systems are related to virulence. PMID- 11437342 TI - Regulation of virulence in Vibrio cholerae. AB - Vibrio cholerae causes the diarrheal disease cholera primarily because it expresses a colonization factor (toxin-coregulated pilus; TCP) and a potent toxin (cholera toxin; CT) within the human intestine. While the true environmental signals that induce CT and TCP expression within the intestine remain unknown, much progress has been made identifying the regulatory factors that modulate their expression. Transcriptional regulation of the genes encoding TCP and CT involves a cascade consisting of a number of regulatory factors located on recently acquired mobile genetic elements as well as others residing within the ancestral Vibrio genome. In vivo studies have revealed interesting differences between the regulation of TCP and CT expression in the laboratory and within the intestine. PMID- 11437343 TI - Regulation of virulence gene expression in Shigella flexneri, a facultative intracellular pathogen. AB - Shigella flexneri and its close relatives are facultative intracellular pathogens of humans and are the etiological agents of bacillary dysentery. These bacteria secrete proteins that enable them to enter human epithelial cells via an elaborate and fascinating cell biology. This behaviour depends on a complicated regulon of virulence genes, whose expression is controlled in response to a multiplicity of environmental signals. This review describes and attempts to interpret these gene control mechanisms. PMID- 11437344 TI - Breaking through the acid barrier: an orchestrated response to proton stress by enteric bacteria. AB - The ability of enteropathogens such as Salmonella and Escherichia coli to adapt and survive acid stress is fundamental to their pathogenesis. Once inside the host, these organisms encounter life-threatening levels of inorganic acid (H+) in the stomach and a combination of inorganic and organic acids (volatile fatty acids) in the small intestine. To combat these stresses, enteric bacteria have evolved elegant, overlapping strategies that involve both constitutive and inducible defense systems. This article reviews the recent progress made in understanding the pH 3 acid tolerance systems of Salmonella and the even more effective pH 2 acid resistance systems of E. coli. Focus is placed on how Salmonella orchestrates acid tolerance by modulating the activities or levels of diverse regulatory proteins in response to pH stress. The result is induction of overlapping arrays of acid shock proteins that protect the cell against acid and other environmental stresses. Most notable among these pH-response regulators are RpoS, Fur, PhoP and OmpR. In addition, we will review three dedicated acid resistance systems of E. coli, not present in Salmonella, that allow this organism to survive extreme (pH 2) acid challenge. PMID- 11437345 TI - Isolation and characterization of a tumor-associated NADH oxidase (tNOX) from the HeLa cell surface. AB - Cell-surface-located, drug-responsive and tumor-associated NADH oxidase (tNOX) proteins were purified and characterized from HeLa cells. The proteins isolated exhibited NADH oxidase activity inhibited by capsaicin and were resistant to heating and to protease digestion. The activity was purified 200- to 500-fold to provide apparently homogeneous gel bands for N-terminal sequencing using three different protocols. All three protocols involved heat (50 degrees C) and proteinase K treatment. Recovery of the total NADH oxidase activity was 86% and inhibition by capsaicin was 60 to 80%. After 450-fold purification, a 52-kDa component was obtained as a single gel band that retained the capsaicin-inhibited NADH oxidase activity. Amino acid composition and partial amino acid sequences were obtained. The partial amino acid sequences were used to generate peptide antisera. Both the peptide antisera and polyclonal antisera to the 52-kDa component immunoprecipitated capsaicin-inhibited NADH oxidase activity and reacted with 52-, 34-, and 17-kDa components on Western blots from different steps of the purification. The tNOX protein exhibited immunological cross reactivity and amino acid sequence identity with tNOX cloned from a HeLa cDNA library using a monoclonal antibody to tNOX from sera of cancer patients. The results provide a direct sequence link between tNOX of the HeLa cell surface and the cloned tNOX representative of patient sera. The tNOX form from the surface of HeLa cells yielded N-terminal sequence consistent with a coidentity of the cell surface and serum forms of the two activities. PMID- 11437346 TI - Macular pigments lutein and zeaxanthin as blue light filters studied in liposomes. AB - Lutein and zeaxanthin are the predominant carotenoids in the human macula lutea. Epidemiological data suggest that an increased intake of a lutein-rich diet correlates with a diminished risk for age-related macular degeneration, a major cause of impaired vision in the elderly. Filtering of blue light has been proposed as a possible mechanism of protection. Here, the blue light filter efficacy of carotenoids was investigated in unilamellar liposomes loaded in the hydrophilic core space with a fluorescent dye, Lucifer yellow, excitable by blue light. Carotenoids were incorporated into the lipophilic membrane. Fluorescence emission in carotenoid-containing liposomes was lower than in carotenoid-free controls when exposed to blue light, indicating a filter effect. Filter efficacy was in the order lutein > zeaxanthin > beta-carotene > lycopene. Some of the difference in blue light filter efficacy of carotenoids is attributable to differences in extinction coefficients, and a major further contribution is suggested to be related to the orientation of the incorporated molecules in the liposomal membrane. PMID- 11437347 TI - Glycosylasparaginase activity requires the alpha-carboxyl group, but not the alpha-amino group, on N(4)-(2-Acetamido-2-deoxy-beta-D-glucopyranosyl)-L asparagine. AB - Glycosylasparaginase catalyzes the hydrolysis of the N-glycosylic bond in N(4)-(2 acetamido-2-deoxy-beta-D-glucopyranosyl)-L-asparagine in the catabolism of N linked oligosaccharides. A deficiency, or absence, of enzyme activity gives rise to aspartylglycosaminuria, the most common disorder of glycoprotein metabolism. The enzyme catalyzes the hydrolysis of a variety of asparagine and aspartyl compounds containing a free alpha-carboxyl group and a free alpha-amino group; computational studies suggest that the alpha-amino group actively participates in the catalytic mechanism. In order to study the importance of the alpha-carboxyl group and the alpha-amino group on the natural substrate to the reaction catalyzed by the enzyme, 14 analogues of the natural substrate were studied where the structure of the aspartyl group of the substrate was changed. The incremental binding energy (DeltaDeltaGb) for those analogues that were substrates was calculated. The results show that the alpha-amino group may be substituted with a group of comparable size, for the alpha-amino group contributes little, if any, to the transition state binding energy of the natural substrate. The alpha-amino group position acts as an "anchor" in the binding site for the substrate. On the other hand, the alpha-carboxyl group is necessary for enzyme activity; removal of the alpha-carboxyl group or changing it to an alpha-carboxamide group results in no hydrolysis reaction. Also, N-acetyl-D-glucosamine is not sufficient for binding to the active site for efficient hydrolysis by the enzyme. These results provide supporting evidence for a proposed intramolecular autoproteolytic activation reaction for the enzyme. However, the results raise a question as to an important role for the alpha-amino group in the catalytic mechanism as indicated in computational studies. PMID- 11437348 TI - RLIP76 is the major ATP-dependent transporter of glutathione-conjugates and doxorubicin in human erythrocytes. AB - We have recently demonstrated that RLIP76, a Ral-binding GTPase activating protein mediates ATP-dependent transport of glutathione (GSH) conjugates of electrophiles (GS-E) as well as doxorubicin (DOX), and that it is identical with DNP-SG ATPase, a GS-E transporter previously characterized by us in erythrocyte membranes (Awasthi et al. Biochemistry 39, 9327-9334). Multidrug resistance associated protein (MRP1) belonging to the family of the ABC-transporters has also been suggested to be a GS-E transporter in human erythrocytes. Using immunological approaches, the present studies were designed to elucidate the relative contributions of RLIP76, MRP1, and P-glycoprotein (Pgp), in the ATP dependent transport of GS-E and DOX in human erythrocytes. In Western blot analyses using antibodies against RLIP76, a strong expression of RLIP76 was observed in erythrocytes. Immunohistochemical studies using a fluorescent probe showed association of RLIP76 with erythrocyte membrane, which was consistent with its transport function. Neither MRP1 nor Pgp were detected in erythrocytes when the antibodies against MRP1 or Pgp were used. In erythrocyte inside-out vesicles (IOVs) coated with antibodies against RLIP76, a dose-dependent inhibition of the ATP-dependent transport of DOX and GS-E, including S-(dinitrophenyl)glutathione (DNP-SG), leukotriene C(4), and the GSH conjugate of 4-hydroxynonenal, was observed with a maximal inhibition of about 70%. On the contrary, in the IOVs coated with the antibodies against MRP1 or Pgp no significant inhibition of the ATP-dependent transport of these compounds was observed. These findings suggest that RLIP76 is the major ATP-dependent transporter of GS-E and DOX in human erythrocytes. PMID- 11437349 TI - A conservative amino acid substitution alters the regiospecificity of CYP94A2, a fatty acid hydroxylase from the plant Vicia sativa. AB - Fatty acid omega-hydroxylation is involved in the biosynthesis of the plant cuticle, formation of plant defense signaling molecules, and possibly in the rapid catabolism of free fatty acids liberated under stress conditions. CYP94A2 is a cytochrome P450-dependent medium-chain fatty acid hydroxylase that was recently isolated from Vicia sativa. Contrary to CYP94A1 and CYP86A1, two other fatty acid hydroxylases previously characterized in V. sativa and Arabidopsis thaliana, CYP94A2 is not a strict omega-hydroxylase, but exhibits chain-length dependent regioselectivity of oxidative attack. Sequence alignments of CYP94A2 with CYP94A1 and molecular modeling studies suggested that F494, located in SRS-6 (substrate recognition site) was involved in substrate recognition and positioning. Indeed, a conservative amino acid substitution at that position markedly altered the regiospecificity of CYP94A2. The observed shift from omega toward omega-1 hydroxylation was prominent with lauric acid as substrate and declined with increasing fatty acid chain length. PMID- 11437350 TI - UDP-galactose 4-epimerase from Escherichia coli: formation of catalytic site during reversible folding. AB - UDP-galactose 4-epimerase from Escherichia coli is a homodimer of molecular weight 39 kDa/subunit having noncovalently bound NAD acting as cofactor. Denaturation by 8 M urea at pH 7.0 causes 85% loss of its secondary structure and dissociation of its constituent molecules. Dilution of the denaturant by buffer at pH 8.5 leads to functional reconstitution of the dimeric holoenzyme. The refolding process is biphasic: after 2 min an equilibrium conformer is formed having 72% of its native secondary structure and by 60 min reactivation becomes complete. The early intermediate has lower energy of activation against thermal denaturation than the reactivated state. Patterns of trypsin digestion suggests a native like structure of this intermediate. Variation of solvent viscosity and ionic strength and inclusion of proline cis-trans isomerase in the refolding process do not alter kinetics of reactivation. Moreover, unaltered kinetics of reactivation against variation of temperature, pH, and duration of denaturation strongly suggests absence of proline cis/trans isomerization. Measurement of kinetics of (i) recovery of tertiary structure by protein fluorescence; (ii) incorporation of NAD from quantitation of bound cofactor; (iii) formation of substrate binding site by specific interaction with extrinsic fluorophore 1 anilino-8-naphthalene sulfonic acid and quenching by 5'-UMP, a competitive inhibitor; and (iv) recovery of activity indicate that they are all comparable. It appears that internal rearrangement of the protein during refolding, shielded from solvent, is the rate-limiting step of generation of cofactor binding site which ultimately leads to maturation of the holoenzyme structure. PMID- 11437351 TI - Purification, characterization, and primary structure of four depressant insect selective neurotoxin analogs from scorpion (Buthus sindicus) venom. AB - Four depressant insect-selective neurotoxin analogs (termed Bs-dprIT1 to 4) from the venom of the scorpion Buthus sindicus were purified to homogeneity in a single step using reverse-phase HPLC. The molecular masses of the purified toxins were 6820.9, 6892.4, 6714.7, and 6657.1 Da, respectively, as determined by mass spectrometry. These long-chain neurotoxins were potent against insects with half lethal dose values of 67, 81, 103, and 78 ng/100 mg larva and 138, 160, 163, and 142 ng/100 mg cockroach, respectively, but were not lethal to mice even at the highest applied dose of 10 microg/20 g mouse. When injected into blowfly larvae (Sarcophaga falculata), Bs-dprIT1 to 4 induced classical manifestations of depressant toxins, i.e., a slow depressant flaccid paralysis. The primary structures of Bs-dprIT 1 to 4 revealed high sequence homology (60-75%) with other depressant insect toxins isolated from scorpion venoms. Despite the high sequence conservation, Bs-dprIT1 to 4 showed some remarkable features such as (i) the presence of methionine (Met(6) in Bs-dprIT1 and Met(24) in Bs-dprIT2 to 4) and histidine (His(53) and His(57) in Bs-dprIT1) residues, i.e., amino acid residues that are uncommon to this type of toxin; (ii) the substitution of two highly conserved tryptophan residues (Trp43 --> Ala and Trp53 --> His) in the sequence of Bs-dprIT1; and (iii) the occurrence of more positively charged amino acid residues at the C-terminal end than in other depressant insect toxins. Multiple sequence alignment, sequence analysis, sequence-based structure prediction, and 3D homology modeling studies revealed a protein fold and secondary structural elements similar to those of other scorpion toxins affecting sodium channel activation. The electrostatic potential calculated on the surface of the predicted 3D model of Bs-dprIT1 revealed a significant positive patch in the region of the toxin that is supposed to bind to the sodium channel. PMID- 11437352 TI - Inhibition of prostaglandin synthesis by nitric oxide in RAW 264.7 macrophages. AB - Multiple effects of nitric oxide (NO) were revealed on the inhibition of prostaglandin (PG) synthesis by a macrophage-like cell line, RAW 264.7 cells, treated with lipopolysaccharide (LPS). NO-generating reagent, N-ethyl-2-(1-ethyl 2-hydroxy-2-nitrosohydrazino)ethanamine (NOC 12), inhibited the release of PG from cells with LPS treatment at higher concentrations although it stimulated the release at 50 microM. PGH synthase (PGHS) activity in the microsome fraction of the LPS-treated cells was inhibited by (+/-)-(E)-methyl-2-[(E)-hydroxyimino]-5 nitro-6-methoxy-3-hexeneamine (NOR 1), another NO-generating reagent, dose dependently. NOC 12 also dose dependently inhibited PG synthesis from exogenous arachidonic acid in those cells. On the other hand, NOC 12 increased PGHS-2 mRNA, while it increased the PGHS-2 protein at concentrations lower than 200 microM or decreased it at higher concentrations. These results suggest that the effect of NO on PGs synthesis in LPS-treated macrophage cells is mainly due to the balance of its stimulations of the transcriptional and/or translational expression of PGHS-2 and the inhibition of the induced PGHS-2 activity. PMID- 11437353 TI - Cloning and characterization of a canine UDP-glucuronosyltransferase. AB - UDP-glucuronosyltransferases (UGTs) are a major family of enzymes catalyzing the transfer of glucuronic acid to a range of endogenous compounds and xenobiotics facilitating their elimination in either urine or bile. Although the dog is commonly used in drug metabolism studies, relatively little is known about the expression and activity of UGTs in this species. This report describes the molecular cloning and functional characterization of the first dog UGT, UGT1A6. The cloned protein is composed of 528 amino acids with the variable region demonstrating a 67-72% identity with the variable regions of mouse, rat, and human UGT1A6. The enzyme expressed stably in V79 cells predominantly catalyzed the glucuronidation of simple, planar phenols (e.g., for 1-naphthol, K(m) = 41 microM, V(max) = 0.07 nmol/min/mg protein), a class of compounds extensively glucuronidated by human UGT1A6. Based on sequence homology and common catalytic activity, this dog UGT1A protein appears to be the canine orthologue of human UGT1A6. PMID- 11437354 TI - Effects of glutaraldehyde polymerization on oxygen transport and redox properties of bovine hemoglobin. AB - Crosslinking of bovine Hb (HbBv) with glutaraldehyde produces a mixture of low oxygen affinity (P(50)) tetrameric and polymeric Hb species (PolyHbBv). Under physiological conditions the P(50) of HbBv and PolyHbBv were 27 and 35 mmHg, respectively. The dependence of the P(50) on pH and chloride ions and the cooperativity (n(50)) of the protein were diminished as a result of glutaraldehyde modification. Rapid kinetic studies showed greater overall rates of oxygen dissociation (k(off)) with little or no change in the association of CO (k(on)) to the modified protein. The rate of nitric oxide (NO)-induced oxidation of the PolyHbBv was slightly lower than that of HbBv. Autoxidation rate of PolyHbBv was 1.4 times faster than that of HbBv. The reaction of hydrogen peroxide (H2O2) with the ferrous (Fe(2+)) and ferric (Fe(3+)) forms of the proteins led to the formation of a more stable ferrylHb (Fe(4+)) in the case of PolyHbBv. Glutaraldehyde polymerization of HbBv alters its normal allosteric mechanisms, autoxidation kinetics and other related redox properties, which may compromise its function and cause greater toxicity when used as an oxygen transport fluid. PMID- 11437355 TI - o-Phthalaldehyde activates the Ca(2+) release mechanism from skeletal muscle sarcoplasmic reticulum. AB - o-Phthalaldehyde (OPA) is a bifunctional reagent that forms an isoindole derivative by reacting with cysteine and lysine residues separated by approximately 0.3 nm. OPA inhibits sarcoplasmic reticulum (SR) Ca(2+)-ATPase activity at low micromolar concentrations and induces Ca(2+) release from actively loaded SR vesicles by activating the ryanodine receptor from fast twitch skeletal muscle. Both ryanodine binding and single-channel activity show a biphasic concentration dependence. At low OPA concentrations (<100 microM), ryanodine binding and single channel activity are stimulated, while at higher concentrations, a time-dependent sequential activation and inhibition of receptor binding is observed. Activation is characterized by a Ca(2+)-independent increase in maximal receptor occupancy. Data are presented to support a model in which Ca(2+) channel and ryanodine binding activity are enhanced due to an intramolecular cross-linking of nearby lysine and nonhyperreactive cysteine residues. OPA complexation with endogenous lysine residue(s) is critical for receptor activation. PMID- 11437356 TI - Identification and quantification of phosphatidylethanolamine-derived glucosylamines and aminoketoses from human erythrocytes--influence of glycation products on lipid peroxidation. AB - While the Maillard reaction of amino acids and proteins as well as its consequences in vivo has been thoroughly investigated, little attention has so far been paid to the glycation of aminophospholipids such as phosphatidylethanolamine (PE) or phosphatidylserine (PS), which are essential for structure and functionality of biological membranes. PE-derived glucosylamines (Schiff-PEs) and aminoketoses (Amadori-PEs) have now for the first time been simultaneously identified and quantified in erythrocytes from diabetics and healthy individuals by liquid chromatography-electrospray mass spectrometry (LC (ESI)MS). The amounts of glycated PE (gPE) were significantly higher in diabetics (0.18-34.2 mol% Schiff-PE and 0.047-0.375 mol% Amadori-PE) than in controls (0.12 3.99 mol% Schiff-PE and 0.018-0.055 mol% Amadori-PE). A positive correlation between fructosylated hemoglobin (HbA(1c)) and the gPE levels was established. No advanced glycation endproducts (AGEs) like 5-hydroxymethylpyrrole-2-carbaldehyde (pyrrole-PE), carboxymethyl (CM-PE), or carboxyethyl (CE-PE) derivatives were detected. To investigate the influence of gPE on lipid peroxidation of biological membranes, liposomes consisting of soy-PE and synthetically prepared Amadori-PE (16:0-16:0) were incubated for several days and the formation of oxidation products was monitored. It could be shown that Amadori-PE extensively promotes lipid peroxidation even in the absence of transition metal ions like Cu(2+) and Fe(3+). Oxidative damage to membrane lipids therefore is supposed to be at least partially caused by the glycation of aminophospholipids. PMID- 11437357 TI - Molecular recognition in the p450cam monooxygenase system: direct monitoring of protein-protein interactions by using optical biosensor. AB - A real-time optical biosensor study on the interactions between putidaredoxin reductase (PdR), putidaredoxin (Pd), and cytochrome P450cam (P450cam) within the P450cam system was conducted. The binary Pd/P450cam and Pd/PdR complexes were revealed and kinetically characterized. The dominant role of electrostatic interactions in formation of productive electron transfer complexes was demonstrated. It was found that Pd/P450cam complex formation and decay obeys biphasic kinetics in contrast to the monophasic one for complexes formed by other redox partners within the system. Evidence for PdR/P450cam complex formation was obtained. It was found that, in contrast to Pd, which binds only to its redox partners, PdR and P450cam were able to form PdR/PdR and P450cam/P450cam complexes. A ternary PdR/Pd/P450cam complex was also registered. Its lifetime was sufficient to permit up to 60 turnovers to occur. The binding of Pd to P450cam and to PdR within the ternary complex occurred at distinct sites, with Pd serving as a bridge between the two proteins. PMID- 11437358 TI - Modulation by cAMP of 1alpha,25-dihydroxyvitamin D3 sensitivity of murine erythroleukemia cells. AB - As we previously reported, 1alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3) dose dependently inhibited not only proliferation of undifferentiated murine erythroleukemia (MEL) cells but also activin A-induced erythroid differentiation of MEL cells. However, the effect of 1,25(OH)2D3 on MEL cell proliferation was significantly greater by one order of magnitude than that on differentiation (IC(50): 9.2 vs 0.8 nM, respectively). The response of activin A-treated mature MEL cells to 1,25(OH)2D3 in the induction of 1,25(OH)2D3-24-hydroxylase (24 OHase) activity, a rapid effect of 1,25(OH)2D3, was enhanced to the same degree as in untreated immature cells, suggesting that differences in capacity of cells to inactivate 1,25(OH)2D3 did not contribute to augmentation of 1,25(OH)2D3 effect in activin A-treated mature cells. Furthermore, neither the number nor the affinity of vitamin D receptors (VDR) differed significantly between activin A treated cells and untreated immature cells. The intracellular cAMP level, which affects 1,25(OH)2D3-mediated induction of 24-OHase activity, was significantly less in activin A-treated mature cells than in immature MEL cells. The addition of dibutyryl cAMP (dbc AMP) to activin A-treated MEL cells dose-dependently attenuated 1,25(OH)2D3-mediated induction of 24-OHase activity, finally to a level comparable to that of the untreated cells at the final concentration of 100 nM dbcAMP, while dbcAMP itself by 100 nM did not affect MEL cell differentiation by 24 h. In summary, we have shown for the first time that 1,25(OH)2D3 exerted its effect on leukemia cells at physiological concentration and that the magnitude of this effect depended on the changes in intracellular cAMP level through stages of differentiation, suggesting that the cAMP-protein kinase A system may be useful as a target for clinical application of vitamin D analogs by improving the sensitivity of leukemic cells to 1,25(OH)2D3. PMID- 11437359 TI - Purification and characterization of glutathione conjugate reductase: a component of the tetrachlorohydroquinone reductive dehalogenase system from Phanerochaete chrysosporium. AB - A membrane-bound glutathione S-transferase and a soluble glutathione conjugate reductase constitute the reductive dehalogenase system of P. chrysosporium. This enzyme system reductively removes chlorine substituents from tetrachlorohydroquinone, a metabolite of pentachlorophenol. The membrane-bound glutathione S-transferase converts tetrachlorohydroquinone to S glutathionyltrichloro-1,4-hydroquinone, which is subsequently reduced to 3,5,6 trichlorohydroquinone by the soluble glutathione conjugate reductase (GCR). This GCR can accept glutathione, dithiothreitol, cysteine, or beta-mercaptoethanol as cosubstrates. GCR was purified to apparent homogeneity by ion-exchange and covalent chromatography. The enzyme exhibits optimum activity at pH 6.0 and 55 degrees C and appears to be a homodimer with a M(r) of approximately 60 kDa. Activity increases as the number of chlorine substituents on the hydroquinone ring is increased. GCR has an apparent K(m) of approximately 33 microM and an apparent k(cat) of approximately 3.43 s(-1) for 2-S-glutathionyl-3,5,6-trichloro 1,4-hydroquinone. Inhibitors of GCR include Cd(2+), Fe(2+), Mn(2+), iodoacetic acid, and p-chloromercuribenzoic acid, suggesting the presence of a catalytic cysteine thiol(s) at the active site. When glutathione is used as a cosubstrate, reduction of S-glutathionyltrichloro-1,4-hydroquinone is accompanied by the production of trichlorohydroquinone and oxidized glutathione in a 1:1 ratio. A mechanism for this novel enzyme is proposed. PMID- 11437360 TI - Certain high molecular weight heparin chains have high affinity for vitronectin. AB - Vitronectin is a 70-kDa protein that is found in both the extracellular matrix as well as serum. Vitronectin is one of the few proteins that regulates both the complement and the coagulation systems. Heparin is known to bind to vitronectin. Review of the literature reveals apparently conflicting outcomes of the interaction of heparin, vitronectin, and the complement system. Previous studies demonstrated that heparin diminishes vitronectin inhibition of complement activity. Numerous studies have also demonstrated that heparin exerts a net inhibitory effect on complement. We used two dimensional affinity resolution electrophoresis (2DARE) to examine this apparent paradox. 2DARE allowed simultaneous determination of binding affinity of heparin for vitronectin as well as the M(r) of the heparin species. In the 2DARE experiment, the interaction of heparin with vitronectin caused retardation of the movement of the heparin through the tube gel in the first dimension. The degree of the retardation of movement was used to calculate the approximate K(d) of that interaction. The heparin from the tube gel was then subjected to a second dimension electrophoresis to determine the M(r) of the heparin. 2DARE analysis of the interaction of heparin with vitronectin clearly demonstrated that a sub population of heparin chains with M(r) > 8000 bound vitronectin with high affinity whereas most high M(r) chains and all lower M(r) chains showed little to no affinity for vitronectin. Our findings are consistent with the hypothesis that a unique binding domain exists in certain heparin chains for vitronectin. PMID- 11437361 TI - Dihydrooxonate is a substrate of dihydroorotate dehydrogenase (DHOD) providing evidence for involvement of cysteine and serine residues in base catalysis. AB - The flavoprotein dihydroorotate dehydrogenase (DHOD) catalyzes the oxidation of dihydroorotate to orotate. Dihydrooxonate is an analogue of dihydroorotate in which the C5 carbon is substituted by a nitrogen atom. We have investigated dihydrooxonate as a substrate of three DHODs, each representing a distinct evolutionary class of the enzyme, namely the two family 1 enzymes from Lactococcus lactis, DHODA and DHODB, and the enzyme from Escherichia coli, which, like the human enzyme, belongs to family 2. Dihydrooxonate was accepted as a substrate although much less efficiently than dihydroorotate. The first half reaction was rate limiting according to pre-steady-state and steady-state kinetics with different electron acceptors. Cysteine and serine have been implicated as active site base residues, which promote substrate oxidation in family 1 and family 2 DHODs, respectively. Mutants of DHODA (C130A) and E. coli DHOD (S175A) have extremely low activity in standard assays with dihydroorotate as substrate, but with dihydrooxonate the mutants display considerable and increasing activity above pH 8.0. Thus, the absence of the active site base residue in the enzymes seems to be compensated for by a lower pK(a) of the 5 position in the substrate. Oxonate, the oxidation product of dihydrooxonate, was a competitive inhibitor versus dihydroorotate, and DHODA was the most sensitive of the three enzymes. DHODA was reinvestigated with respect to product inhibition by orotate. The results suggest a classical one-site ping-pong mechanism with fumarate as electron acceptor, while the kinetics with ferricyanide is highly dependent on the detailed reaction conditions. PMID- 11437362 TI - Effects of acyclo-retinoic acid and lycopene on activation of the retinoic acid receptor and proliferation of mammary cancer cells. AB - The biochemical mechanisms underlying the inhibitory effects of lycopene, the main tomato carotenoid, on the growth of cancer cells are largely unknown. It has been hypothesized that lycopene derivatives may act as ligands for a nuclear receptor in analogy to retinoic acid, the hormone derived from beta-carotene. The inhibition of human mammary cancer (MCF-7) cell growth and the transactivation of the retinoic acid receptor (RAR) reporter gene by synthetic acyclo-retinoic acid, the open chain analog of retinoic acid, was compared to the effects of lycopene and retinoic acid in the same systems. Acyclo-retinoic acid activated the DR-5 retinoic acid response element with a approximately 100-fold lower potency than retinoic acid. This effect was independent of cotransfection with the RARalpha receptor. Lycopene exhibited only very modest activity in this system. In contrast to the results from the transactivation studies, acyclo-retinoic acid, retinoic acid, and lycopene inhibited cell growth with a similar potency. Preincubation with each of the three compounds slowed down cell cycle progression from G1 to S phase. In summary, acyclo-retinoic acid inhibited cancer cell growth and interacted with RAR. However, it exhibited low affinity for RAR and a correspondingly low efficacy in activating this receptor, indicating that RAR does not mediate the growth inhibitory effect of the compound. In addition, the concentrations of acyclo-retinoic acid and of lycopene required for inducing inhibition of cell growth were similar, suggesting that acyclo-retinoic acid is unlikely to be the active metabolite of lycopene. PMID- 11437363 TI - Suppression of protein kinase C signaling by the novel isoform for bovine PGF(2alpha) receptor. AB - A cDNA clone for a novel isoform of prostaglandin (PG) F(2alpha) receptor (FP) was isolated from the cDNA pool of the bovine corpus luteum. The sequence analysis revealed that the new FP isoform (FP(a)) encodes a 295-amino acid protein carrying a specific 28-amino acid sequence from the middle of transmembrane segment VI to the carboxyl terminus. Because only one copy gene has been identified for FP, FP(a) was generated by alternative mRNA splicing at the middle of the VI transmembrane region, resulting in the lack of a VII transmembrane segment and an intracellular carboxyl tail. The RT-PCR analysis for FP and FP(a) indicated that both mRNAs are expressed similarly during the estrous cycle and pregnancy. The PGF(2alpha) stimulation drastically enhanced protein kinase C (PKC) activity in the COS-7 cell transfected with FP, whereas no PKC activation was detected in FP(a)-transfected cells. Cotransfection of an excess amount of FP(a) markedly reduced FP-mediated PKC activity, suggesting that the novel FP isoform might play a role as a negative regulator to attenuate normal FP function. PMID- 11437364 TI - Force measurement and inhibitor binding assay of monomer and engineered dimer of bovine carbonic anhydrase B. AB - We applied atomic force microscopy (AFM) to study the intramolecular mechanics of the globular protein molecule, bovine carbonic anhydrase B. The immobilized protein on an amino-functionalized silicon wafer was pulled from its N- and C termini after being covalently cross-linked to the AFM tip, and the relationship between the tensile force applied on the protein and its extension was recorded. The native enzyme (having 261 residues with two Cys added at its ends, and in a theoretical stretching length of 96 nm) was extended only to 13 +/- 2 nm under physiological conditions before disruption of the covalent cross-linking system. Contrary to the above observation, an engineered dimer was extended to about 110 nm even in the absence of the denaturant. The difference was ascribed to the presence or presumed absence of a "knot" structure at the C-terminal end of the two forms, respectively. When a specific inhibitor was added to the experimental solution, native monomers (sp activity = 88% of the wild type enzyme) were extended to 28 +/- 4 nm, whereas dimers (sp activity = 46%) were extended to about 56 +/- 3 nm, suggesting that both monomeric units in the dimer could bind inhibitor molecules, which was further corroborated by a titration experiment using a fluorescent inhibitor. Thus, one of the monomeric units in the engineered dimer was concluded to be enzymatically inactive but capable of binding inhibitors. PMID- 11437365 TI - Remnant lipoproteins induce endothelial plasminogen activator inhibitor-1. AB - Remnant lipoproteins (RLPs) accumulate in type III hyperlipoproteinemia, a condition associated with significant cardiovascular morbidity. The effect of RLPs on fibrinolysis is unknown. Our aim was to study the effect of RLPs on endothelial expression of plasminogen activator inhibitor-1 (PAI-1). After 24-h culture of human aortic endothelial cells with RLPs at concentrations of 0 (control), 0.038, or 0.076 mg triglyceride/mL, postculture PAI-1 antigen concentrations were: 870 +/- 80, 1963 +/- 183 (P = 0.005), and 3551 +/- 177 ng/mL (P < 0.001), respectively. Furthermore, after 24-h incubation of endothelial cells with RLPs (0 or 0.076 mg triglyceride/mL), PAI-1 activity increased from 0.667 +/- 0.144 to 1.268 +/- 0.198 U/mL, respectively (P = 0.008) and endothelial PAI-1 mRNA increased to 2.7 +/- 0.66 that of control (P = 0.048). In conclusion, RLPs from patients with type III hyperlipoproteinemia induce endothelial cell PAI 1 expression, which may contribute to a prothrombotic state. PMID- 11437366 TI - Role of cysteine-291 and cysteine-322 in the polymerization of human tau into Alzheimer-like filaments. AB - Filamentous tau pathology is central to a large number of dementing disorders, including Alzheimer's disease in which polymerized tau is hyperphosphorylated. Previous studies on heparin-dependent tau polymerization, using recombinant tau isoforms lacking Cys-291, suggest that tau dimerization via Cys-322 is critical for initiation of assembly of soluble tau into filaments. We report heparin dependent in vitro polymerization of human recombinant tau (1-383 isoform), containing both Cys-291 and Cys-322, into paired helical filaments as characterized by electron microscopy. Tau polymerization, under physiological tau concentrations in the presence of dithiothreitol (DTT), was followed by a Thioflavine S fluorescence assay. To understand the molecular basis for heparin induced tau polymerization, we expressed and purified C291A, C322A, and C291A/C322A tau mutants. The DTT requirement for tau polymerization was abolished using either the C291A or C322A tau mutant and polymerization was not observed with the C291A/C322A tau double mutant. Analysis by sodium dodecyl sulfate gel electrophoresis showed that, unlike wild type tau, a significant amount of the C291A mutant and the C322A mutant is present as a disulfide bonded dimer. Taken together these results suggest that, in isoforms containing both Cys-291 and Cys 322, a dimeric tau with an intermolecular disulfide bond through either Cys-291 or Cys-322 is presumably acting as a seed for initiation of tau polymerization. PMID- 11437367 TI - Poly(ADP-ribosyl)ation of transcription factor Yin Yang 1 under conditions of DNA damage. AB - Under conditions of severe DNA damage the nuclear enzyme poly(ADP-ribose) polymerase 1 (PARP-1) is activated, catalyzing the modification of proteins by forming and attaching to them poly(ADP-ribose) chains. A specific physical interaction between PARP-1 and transcription factor Yin Yang 1 (YY1) in vitro was shown previously, which had important consequences for the activities of both proteins. It is demonstrated here that YY1 and PARP-1 form complexes in vivo. YY1 was transiently poly(ADP-ribosyl)ated immediately after genotoxic treatment of HeLa cells. The narrow time frame of the modification coincides with that known for the activation of PARP-1 under these conditions. This immediate modification correlated with a decreased affinity of YY1 to its cognate DNA binding sites. PMID- 11437368 TI - Oxidative hydrolysis of scoparone by cytochrome p450 CYP2C29 reveals a novel metabolite. AB - Regioselective 7-demethylation of scoparone is regularly employed as an indicator of phenobarbital-like induction of rat liver cytochrome P450 isoform CYP2B1, e.g., by the antiepileptic drug phenytoin. After induction with phenobarbital and phenytoin, a new reaction sequence catalyzed by Cyp2c29 was identified in mouse liver microsomes. Cyp2c29-dependent 6-demethylation of scoparone resulted in the formation of isoscopoletin, an intermediate which is susceptible to further oxidation. This subsequent oxidation was also catalyzed by Cyp2c29 with a K(m) of 30,31 microM and a V(max) of 3,41 microM/min x microM P450, and resulted in the formation of the new metabolite 3-[4-methoxy-p-(3,6)-benzoquinone]-2-propenoate. This novel metabolite is the product of two consecutive oxidation reactions, proceeding over isoscopoletin to a putative lactone which is accessible to immediate hydrolysis, due to the onium character of the ring oxygen. This opening of the lactone ring corresponds to an oxidative hydrolysis. Differential oxidation of scoparone can be used as a sensitive indicator for distinguishing between different cytochrome P450 isoforms. PMID- 11437369 TI - Dual mechanisms of 9-beta-D-arabinofuranosylguanine resistance in CEM T lymphoblast leukemia cells. AB - The guanine nucleoside analog araG is selectively toxic to T-lymphoblasts and has recently shown promise in treatment of lymphoid malignancies of T-cell origin. The molecular mechanism of this tissue-selective cytotoxicity is, however, yet unclear. AraG is phosphorylated, and thereby pharmacologically activated, by the mitochondrial deoxguanosine kinase and the cytosolic/nuclear deoxycytidine kinase. We have recently shown that araG is predominantly incorporated into mitochondrial DNA of cancer cell lines, which suggests a role of mitochondria as its pharmacological target. In the present study, we have generated araG resistant CEM T-lymphoblast cell lines and show that araG resistance may occur by two separate molecular mechanisms that can occur sequentially. The first mechanism is associated with a decrease of araG incorporation into mitochondrial DNA, and the second event is associated with loss of dCK activity. PMID- 11437370 TI - Inhibition of Na,K-ATPase activates PI3 kinase and inhibits apoptosis in LLC-PK1 cells. AB - In the present study we used LLC-PK1 cells, a porcine renal proximal tubular cell line, to investigate whether PI3 kinase activation was involved in the anti apoptotic effect of ouabain, a specific inhibitor of Na,K-ATPase. Apoptosis was induced by actinomycin D (Act D, 5 microM) and assessed by appearance of hypodiploid nuclei and DNA fragmentation. Ouabain attenuated Act D-induced apoptotic response in a dose-dependent manner. Incubation in a low K(+) medium (0.1 mM) which is another way to decrease Na,K-ATPase activity also had anti apoptotic effect. Both ouabain and low K(+) medium increased the PI3 kinase activity in p85 immunoprecipitates. Ouabain, as well as incubation in the low K(+) medium, also increased the phosphorylation of Akt. Inhibition of PI3 kinase by either wortmannin or LY294002 reversed the cytoprotective effect of ouabain. These data together indicate that inhibition of Na,K-ATPase activates PI3 kinase in LLC-PK1 cells which could then exert the cytoprotective effect. PMID- 11437371 TI - Analysis of type 1 ryanodine receptor-12 kDa FK506-binding protein interaction. AB - Although dissociation of the 12 kDa FK506 binding protein (FKBP12)-type 1 ryanodine receptor (RyR1) complex by macrolide immunosuppressants is well documented, effects of many solutes and drugs have not been quantitated. In the current study, the influence of these on binding between solubilised RyR1 and an FKBP12-glutathione-S-transferase fusion protein was analysed using a novel assay. Association between these two proteins is stable, and is not greatly altered by changes in temperature, pH, cations, and endogenous solutes over physiological ranges. Ascomycin, an FK506 analogue, was identified for the first time as a drug which can disrupt the FKBP12-RyR1 complex. PMID- 11437372 TI - Fluorescence anisotropy: a method for early detection of Alzheimer beta-peptide (Abeta) aggregation. AB - Time-resolved anisotropy measurements (TRAMS) have been used to study the aggregation of the beta-amyloid (Abeta) peptide which is suspected of playing a central role in the pathogenesis of Alzheimer's Disease (AD). The experiments, which employ small quantities of fluorescently-labelled Abeta, in addition to the untagged peptide, have shown that the sensitive TRAMS technique detects the presence of preformed "seed" particles in freshly prepared solutions of Abeta. More importantly, as 100 microM solutions of Abeta containing tagged Abeta at a concentration level of either 0.5 or 1 microM are incubated, the TRAMS prove capable of detection of the peptide aggregation process through the appearance of a continuously increasing "residual anisotropy" within the time-resolved fluorescence data. The method detects Abeta aggregation in its earliest stages, well before complexation becomes apparent in more conventional methods such as the thioflavin T fluorescence assay. The TRAMS approach promises to provide a most attractive route for establishment of a high-throughput procedure for the early detection of the presence of amyloid aggregates in the screening of biological samples. PMID- 11437373 TI - High incidence of antinuclear antibodies that recognize the matrix attachment region. AB - The matrix attachment region (MAR) is a distinctive genomic DNA involved in a variety of nuclear processes through association with the nuclear matrix. Recent studies suggest that nuclear matrix is altered in the process of apoptosis and presented to the immune system, leading to the production of autoantibodies against its protein components. To see whether MARs are also recognized by autoantibodies, a collection of human sera containing antinuclear antibodies was screened for the presence of binding activities against cloned MARs. We found that MAR-binding activities are quite common in these sera. There was a positive correlation among the MAR-binding titers for three different MAR probes. As expected, the MAR-binding activity was copurified with serum IgG, and subclass analysis with affinity-purified IgG on MAR-Sepharose showed a predominance of IgG2 isotype. Several lines of evidence implied that the anti-MAR antibodies detected here is distinct from the ordinary anti-DNA antibodies that are reactive to bulk DNA. PMID- 11437374 TI - Testosterone prevents orchidectomy-induced bone loss in estrogen receptor-alpha knockout mice. AB - To examine the role of the estrogen receptor-alpha (ERalpha) during male skeletal development, bone density and structure of aged ERalphaKO mice and wild-type (WT) littermates were analyzed and skeletal changes in response to sex steroid deficiency and replacement were also studied. In comparison to WT, ERalphaKO mice had smaller and thinner bones, arguing for a direct role of ERalpha to obtain full skeletal size in male mice. However, male ERalphaKO mice had significantly more trabecular bone as assessed both by pQCT and histomorphometry, indicating that ERalpha is not essential to maintain cancellous bone mass. Six weeks following orchidectomy (ORX), both WT and ERalphaKO mice showed high-turnover osteoporosis as revealed by increases in serum osteocalcin and decreases in trabecular (-38% and -58% in WT and ERalphaKO, respectively) and cortical bone density (-5% and -4% in WT and ERalphaKO, respectively). Administration of testosterone propionate (T, 5 mg/kg/day) completely prevented bone loss both in ERalphaKO and in WT mice. As expected, estradiol (E2, 60 microg/kg/day) replacement did not prevent cancellous bone loss in ORX ERalphaKO mice. However, E2 stimulated bone formation at the endocortical surface in ORX ERalphaKO, suggesting that osteoblasts may respond to nonERalpha-mediated estrogen action. In conclusion, although functional ERalpha may play a significant role during male skeletal development, this receptor does not seem essential for androgen mediated skeletal maintenance in older male mice. PMID- 11437375 TI - Structural Insights into Cdk5 activation by a neuronal Cdk5 activator. AB - Although Cdk5 shows high sequence identity to Cdk1 and Cdk2, it can be fully activated by its neuronal activators p35/p25(nck5a) and p39(nck5ai) in a phosphorylation-independent manner. To understand structural basis of the Cdk5/p25(nck5a) activation, the complex is modelled to assume either an obstructed or an opened conformation based on X-ray structures of the unphosphorylated or the phosphorylated Cdk2/cyclin A complex, respectively. Comparison and analysis of the two models, along with mutagenesis studies of p25(nck5a), suggest that the opened form represents more closely the structure of active Cdk5/p25(nck5a). The results provide a rationale basis for understanding the phosphorylation-independent activation of Cdk5/p25(nck5a). PMID- 11437376 TI - Overexpression of EC-SOD suppresses endothelial-cell-mediated LDL oxidation. AB - Reactive oxygen species have been proposed to play important roles in atherosclerosis. To investigate the protective role of extracellular superoxide dismutase (EC-SOD), its inhibition of endothelial-cell-mediated LDL oxidation was examined. We constructed the recombinant adenovirus AxCAEC-SOD expressing human EC-SOD by CAG promoter. Infection of endothelial cells with AxCAEC-SOD resulted in EC-SOD protein secretion in a dose-dependent manner and a decrease of endothelial-cell-derived superoxide production. Moreover, it was proven to coexist with heparan sulfate by immunohistochemical staining. Endothelial-cell mediated LDL oxidation enhanced by ferric-sodium EDTA was inhibited by 47% in TBARS formation by AxCAEC-SOD infection. In agarose gel electrophoresis, AxCAEC SOD decreased the negative charge of oxidized LDL by 50% and suppressed fragmentation of apolipoprotein B. These results suggested that human EC-SOD localized in the extracellular space and reduced endothelial-cell-mediated LDL oxidation. In subendothelial space, EC-SOD bound on heparan sulfate might suppress LDL oxidation through reduction of superoxide anion. PMID- 11437377 TI - Strand exchange reaction in vitro and DNA-dependent ATPase activity of recombinant LIM15/DMC1 and RAD51 proteins from Coprinus cinereus. AB - We previously cloned recA-homolog genes from a basidiomycete, Coprinus cinereus, and obtained the recombinant proteins (Nara et al., Mol. Gen. Genet. 262, 781 789, 1999, see Ref. 1; Nara and Sakaguchi, Biochem. Biophys. Res. Commun. 275, 97 102, 2000, see Ref. 2). The primary purpose of the present study was to characterize the biochemical properties of the recombinant LIM15/DMC1 (CoLIM15) and RAD51 (CoRAD51) proteins. We purified the recombinant proteins, and their molecular masses were 37 and 35 kDa, respectively. Both enzymes showed DNA dependent ATPase activity and ATP-dependent strand exchange reaction in vitro. CoRad51 was a five- to sixfold stronger DNA-dependent ATPase and showed greater dependency on single-stranded DNA than CoLim15. In meiosis, both enzymes were highly accumulated in the meiotic tissue at leptotene and zygotene stages at which the homologous chromosomes pair, but disappeared just before the pachytene stage at which they recombine. From these and the previously reported results, we discuss here the relationships between the enzymes and meiosis. PMID- 11437378 TI - Obtusifoliol 14alpha-demethylase (CYP51) antisense Arabidopsis shows slow growth and long life. AB - Obtusifoliol 14alpha-demethylase is a plant orthologue of sterol 14alpha demethylase (CYP51) essential in sterol biosynthesis. We have prepared CYP51 antisense Arabidopsis in order to shed light on the sterol and steroid hormone biosynthesis in plants. Arabidopsis putative CYP51 cDNA (AtCYP51) was obtained from Arabidopsis expressed sequence tag (EST) library and its function was examined in a yeast lanosterol 14alpha-demethylase (Erg11) deficient mutant. A recombinant AtCYP51 protein fused with a yeast Erg11 signal-anchor peptide was able to complement the erg11 mutation, which confirmed AtCYP51 to be a functional sterol 14alpha-demethylase. AtCYP51 was then used to generate transgenic Arabidopsis by transforming with pBI vector harboring AtCYP51 in the antisense direction under CaMV35S promoter. The resulting transgenic plants were decreased in accumulation of AtCYP51 mRNA and increased in the amount of endogenous obtusifoliol. They showed a semidwarf phenotype in the early growth stage and a longer life span than control plants. This newly found phenotype is different from previously characterized brassinosteroid (BR)-deficient campesterol biosynthesis mutants. PMID- 11437379 TI - Molecular cloning, developmental expression, and hormonal regulation of zebrafish (Danio rerio) beta crystallin B1, a member of the superfamily of beta crystallin proteins. AB - The cDNA sequence of beta crystallin B1 was determined from zebrafish (Danio rerio) and compared to the corresponding genes of bovine, rat, chicken, human, and Xenopus. Multispecies comparison of superfamily diversity demonstrated beta crystallin B1 homology between zebrafish, bovine, chicken, and rat, but large distances to beta crystallin B2 and B3. Zebrafish cDNA has a size of 943 nucleotides and encodes a polypeptide of 233 amino acids. Zebrafish beta crystallin B1 shares 71.30, 75.86, and 71.00% similarities with bovine, chicken, and rat beta crystallin B1, respectively. Northern blot analysis revealed a single 0.9-kb beta crystallin B1 transcript which was expressed and progressively increased in the first 20 h of zebrafish embryogenesis. Whole-mount in situ hybridization revealed that the beta crystallin B1 transcript was only specifically expressed in the lens region of the eye. A starvation experiment revealed no variation in mRNA levels after 14 and 21 days. An experiment in which hormone was injected showed that the beta crystallin B1 transcript first increased 24 h after the injection of insulin-like growth factor I, insulin-like growth factor II, or growth hormone, then decreased 48 h after injection. The beta crystallin B1 transcript continuously increased after insulin was injected. Taken together, our results identify the early specific expression of beta crystallin B1 within the lens. Despite small differences, these results indicate that both the structure of the beta crystallin B1 protein and its involvement with regulation by growth factors appear to have been remarkably conserved. PMID- 11437380 TI - Functional characterization of the human multidrug transporter, ABCG2, expressed in insect cells. AB - ABCG2 (also called MXR (3), BCRP (4), or ABCP (5) is a recently-identified ABC half-transporter, which causes multidrug resistance in cancer. Here we report that the expression of the ABCG2 protein in Sf9 insect cells resulted in a high capacity, vanadate-sensitive ATPase activity in isolated membrane preparations. ABCG2 was expressed underglycosylated, and its ATPase activity was stimulated by daunorubicin, doxorubicin, mitoxantrone, prazosin and rhodamine 123, compounds known to be transported by this protein. ABCG2-ATPase was inhibited by low concentrations of Na-orthovanadate, N-ethylmaleimide and cyclosporin A. Verapamil had no effect, while Fumitremorgin C, reversing ABCG2-dependent cancer drug resistance, strongly inhibited this ATPase activity. The functional expression of ABCG2 in this heterologous system indicates that no additional partner protein is required for the activity of this multidrug transporter, probably working as a homodimer. We suggest that the Sf9 cell membrane ATPase system is an efficient tool for examining the interactions of ABCG2 with pharmacological agents. PMID- 11437381 TI - Tumor autocrine motility factor is an angiogenic factor that stimulates endothelial cell motility. AB - Autocrine motility factor (AMF) is a type of tumor-secreted cytokine which primarily stimulates tumor cell motility via receptor-mediated signaling pathways, and is thought to be connected to tumor progression and metastasis. Using in vivo models, we showed that critical neovascularization responded to a biological amount of AMF. This angiogenic activity was fixed by specific inhibitors against AMF. AMF stimulated in vitro motility of human umbilical vein endothelial cells (HUVECs), inducing the expression of cell surface AMF receptor localizing a single predominant perinuclear pattern closely correlated with its motile ability. AMF also elicited the formation of tube-like structures mimicking angiogenesis when HUVECs were grown in three-dimensional type I collagen gels. We further immunohistochemically detected AMF receptors on the surrounding sites of newborn microvessels. These findings suggest that AMF is a possible tumor progressive angiogenic factor which may act in a paracrine manner for the endothelial cells in the clinical neoplasm, and it will be a new target for antiangiogenic treatment. PMID- 11437382 TI - Induction of disabled-2 gene during megakaryocyte differentiation of k562 cells. AB - Megakaryocyte differentiation is often accompanied by the changes of gene expression pattern. Here we reported that the expression of DAB2, a putative adaptor protein in cell signaling, was induced at the protein and mRNA levels upon 12-O-tetradecanoylphorbol-13-acetate-mediated megakaryocyte differentiation of human chronic myeloid leukemic K562 cells. On the other hand, the differentiation agents DMSO and retinoic acid had no effect on DAB2 expression. Analysis of promoter activity with the human DAB2 luciferase reporter constructs suggested that the regulation is partially at the transcriptional level. The responsive sequences located within an 80-bp DAB2 promoter region. To determine the involvement of MEK1-p42/p44 MAPK pathway in mediating DAB2 gene expression, we have performed the following experiments and found that (i) there was sustained activation of p42/p44 MAPK, but not p38 MAPK, upon K562 cells differentiation; (ii) application of MEK1 inhibitor U0126 reduced the expression of DAB2 protein, mRNA and promoter activity, as well as cell differentiation; (iii) constitutively active MEK1 increased DAB2 promoter activity; and (iv) dominant negative ERK2 abolished constitutively active MEK1-induced DAB2 promoter activity. Taken together, our results indicate that DAB2 gene is induced upon megakaryocyte differentiation by the MEK1-p42/p44 MAPK pathway and may define a new role of DAB2 in hematopoietic cell differentiation. PMID- 11437383 TI - Glycine-extended gastrin promotes the invasiveness of human colon cancer cells. AB - Colorectal cancers express significant amounts of immature glycine-extended gastrin (G-Gly) and G-Gly is able to stimulate cell proliferation in colonic cell lines and mucosa. Here we wished to investigate whether G17-Gly promote the invasiveness of LoVo human colonic cancer cells, a process which requires degradation of extracellular matrix by proteases and concomitant induction of cell migration. We confirmed that LoVo cells express gastrin and gastrin/CCK-B receptor mRNAs. We showed that these cells secrete matrix metalloproteinase (MMP) 1, -2, and -9. The function of MMP being to degrade components of extracellular matrix, they may thus favor cell migration. As compared to controls, G17-Gly (10( 7) to 10(-12) M) significantly enhanced about two to three times the LoVo cell migration through Matrigel, an artificial basement matrix barrier. Moreover, G17 Gly increased and gastrin/CCK-B receptor antagonists decreased MMP secretion in conditioned culture media of LoVo cells. Our findings show that physiological doses of incompletely processed form of gastrin induce the invasiveness of tumor cells in vitro and suggest a novel potential role for this peptide in the metastatic process of colonic cancers in vivo. PMID- 11437384 TI - Ni(2+), a double-acting inhibitor of neuronal nitric oxide synthase interfering with L-arginine binding and Ca(2+)/calmodulin-dependent enzyme activation. AB - Ni(2+), a toxic and carcinogenic pollutant and one of the leading causes of contact dermatitis, is shown to inhibit neuronal nitric oxide synthase (nNOS) in a competitive, reversible manner with respect to the substrate l-arginine (K(i) = 30 +/- 4 microM). The IC(50) values were dependent on calmodulin (CaM) concentration, but proved independent of Ca(2+), tetrahydrobiopterin (BH(4)) and other essential cofactors. Ni(2+) also inhibited CaM-dependent cytochrome c reduction, NADPH oxidation, and H(2)O(2) production by nNOS. Overall, the action profile of Ni(2+) was suggestive of an unusual, double-acting inhibitor of nNOS affecting l-arginine-binding and Ca(2+)/CaM-dependent enzyme activation. PMID- 11437385 TI - Identification of coding single-nucleotide polymorphisms in human taste receptor genes involving bitter tasting. AB - T2Rs comprise a G-protein-coupled receptor superfamily that contains functionally defined bitter taste receptors. Here we report the tissue expressions and coding single-nucleotide polymorphisms (cSNPs) in human T2R genes (hT2R3, hT2R4, and hT2R5) on chromosome 7q31. We first demonstrated that hT2R3, hT2R4, and hT2R5 are actually expressed in the circumvallate papillae of the human tongue by reverse transcription-polymerase chain reaction (RT-PCR). We identified six cSNPs within the T2R receptor genes. The hT2R4 and hT2R5 contained four and one cSNPs that cause missense mutations, respectively, while hT2R3 included one silent nucleotide mutation. However, we could not find any nonsense mutations that resulted in a frameshift or a premature stop codon within the open reading frames. Genotype frequencies of each cSNP were in Hardy-Weinberg equilibrium. The identification of nucleotide diversity and amino acid polymorphisms in human T2R receptors could help clarify individual differences in the acceptability and sensitivity to bitter compounds. PMID- 11437386 TI - Use of an in silico approach to define the gene structure of eukaryotic adenylyl cyclases. AB - The limited information available regarding the gene structure of adenylyl cyclases (AC), which catalyze the synthesis of cAMP, suggests a complex arrangement with many exons and large introns such that molecular techniques to define these gene structures are time- and labor-intensive. We report here the use of a computer-based approach involving the assembly of fragmented sequence data generated by the Human Genome Project and nucleic acid analysis software to decipher the gene structure of human and murine AC 6 and other human AC isoforms (ACs 3, 7, and 8). The results, which document 21 exons in human and murine AC 6, human AC 3, 18 exons in AC 8, and 24 exons in AC 7, show substantial conservation of exon organization in the AC family and in particular regions of the AC protein. Application of such in silico methods should prove useful to characterize genes for other ACs and protein families and data provided here should facilitate studies of polymorphisms in AC genes. PMID- 11437387 TI - Inteins as enzymes. AB - The term "self-catalyzed" as applied to protein processing reactions might be considered a contradiction, since catalysis implies that the catalyst is regenerated without change. However, as our understanding of protein autoprocessing reactions such as protein splicing advances, it is becoming clear that they have many of the hallmarks of enzymatic reactions. In this review, we will examine the properties of protein splicing elements, or inteins, and show how these can be understood in terms of enzyme catalysis, both with respect to substrate specificity and the stabilization of reactive intermediates. PMID- 11437388 TI - Photodynamic activity of 5,10,15,20-tetrakis(4-methoxyphenyl)porphyrin on the Hep 2 human carcinoma cell line: effect of light dose and wavelength range. AB - The photodynamic activity of 5,10,15,20-tetrakis(4-methoxyphenyl)porphyrin (TMP) has been investigated in two systems: reverse micelles of n-heptane/sodium bis(2 ethylhexyl)sulfosuccinate (AOT)/water-bearing photooxidizable substrates and on a Hep-2 human carcinoma cell line. The effect of variation in the light dose and wavelength range (360-800, 455-800, and 590-800 nm) was compared in both media. The aerobic singlet oxygen-mediated photooxidation of L-tryptophan (Trp) was used as a model of biological substrate in a micellar system. A considerable increase of the observed rate constants of Trp (k(Trp)(obs)) was noted, increasing the irradiated area of the TMP spectrum. In vitro, the survival curves of Hep-2 cells, treated with TMP, were markedly dependent on the light wavelength ranges used for irradiation. A linear behavior between k(Trp)(obs) and the photoinactivation rate of Hep-2 cells was found, indicating that the singlet oxygen (1O2 ) is the main species responsible for cell inactivation. These results contributed to an understanding of the photodynamic process yielded by this porphyrin in vitro and the sensitivity of Hep-2 cells to photodamage. PMID- 11437389 TI - Inhibition of beta-lactamases by 6,6-bis(hydroxylmethyl)penicillanate. AB - beta-Lactamases of classes A and C are the two most prevalent resistant determinants to beta-lactam antibiotics among bacterial pathogens. Both these enzymes pursue different mechanisms for their catalytic processes, highlighted by the fact that the hydrolytic water molecule in each approaches the ester of the intermediary acyl-enzyme species from the opposite ends. 6,6 Bis(hydroxylmethyl)penicillanate was designed as an inhibitor that would impair the approach of the hydrolytic water molecule in either of these enzymes upon formation of the acyl-enzyme species. The design, synthesis, and kinetic evaluation of this inhibitor are disclosed herein. PMID- 11437390 TI - Effect of an E461G mutation of beta-galactosidase (Escherichia coli, lac Z) on pL rate profiles and solvent deuterium isotope effects. AB - An E461G mutation of beta-galactosidase results in the disappearance of the high pL (L = H, D) downward break in the rate profiles for k(cat)/K(m) for wild-type enzyme-catalyzed hydrolysis of 4-nitrophenyl beta-D-galactopyranoside (Gal-OPNP) and a decrease from (k(cat))(HOH)/(k(cat))(DOD) = 1.7 to (k(cat))(HOH)/(k(cat))(DOD) = 1.2 in the solvent deuterium isotope effect. These observations provide evidence that the propionic acid side chain of Glu 461 is protonated at catalytically active free beta-galactosidase and they are consistent with a role for this residue in Bronsted acid catalysis at the leaving group. The earlier observation that this same E461G mutation results in the loss of a downward break at high pH in the rate profile for k(s) for transfer of the beta-D-galactopyranosyl group from beta-galactosidase to water cannot be simply explained by a mechanism in which the single side chain of Glu 461 functions to provide general acid catalysis in the rate limiting step for formation of the beta-D-galactopyranosyl intermediate and general base catalysis of breakdown of this intermediate. Evidence is presented that there may be different catalytic mechanisms, with different roles for the side chain for Glu-461, for nucleophilic addition of water and of small alkyl alcohols to the beta-D-galactopyranosyl reaction intermediate. PMID- 11437391 TI - Effect of ginger constituents and synthetic analogues on cyclooxygenase-2 enzyme in intact cells. AB - Seventeen pungent oleoresin principles of ginger (Zingiber officinale, Roscoe) and synthetic analogues were evaluated for inhibition of cyclooxygenase-2 (COX-2) enzyme activity in the intact cell. These compounds exhibited a concentration and structure dependent inhibition of the enzyme, with IC(50) values in the range of 1-25 microM. Ginger constituents, [8]-paradol and [8]-shogaol, as well as two synthetic analogues, 3-hydroxy-1-(4-hydroxy-3-methoxyphenyl)decane and 5-hydroxy 1-(4-hydroxy-3-methoxyphenyl)dodecane, showed strong inhibitory effects on COX-2 enzyme activity. The SAR analysis of these phenolic compounds revealed three important structural features that affect COX-2 inhibition: (i) lipophilicity of the alkyl side chain, (ii) substitution pattern of hydroxy and carbonyl groups on the side chain, and (iii) substitution pattern of hydroxy and methoxy groups on the aromatic moiety. PMID- 11437392 TI - The specificity of methyl transferases involved in trans mycolic acid biosynthesis in Mycobacterium tuberculosis and Mycobacterium smegmatis. AB - Trans mycolic acid content is directly related to cell wall fluidity and permeability in mycobacteria. Carbon-13 NMR spectroscopy of mycolic acids isolated from Mycobacterium tuberculosis (MTB) and Mycobacterium smegmatis (MSM) fed 13C-labeled precursor molecules was used to probe the biosynthetic pathways that modify mycolic acids. Heteronuclear correlation spectroscopy (HMQC) of ketomycolic acid from MTB allowed assignment of the complete 13C-NMR spectrum. Incorporation patterns from [1-13C]-acetate and [2-13C]-acetate feeding experiments suggested that the mero chain and alpha branch of mycolic acids are both synthesized by standard fatty acid biosynthetic reactions. [13C-methyl]-L methionine was used to specifically label carbon atoms derived from the action of the methyl transferases involved in meromycolate modification. To enrich for trans mycolic acids a strain of MTB overexpressing the mma1 gene was labeled. Carbon-carbon coupling was observed in mycolate samples doubly labeled with 13C acetate and [13C-methyl]-L-methionine and this information was used to assess positional specificity of methyl transfer. In MTB such methyl groups were found to occur exclusively on carbons derived from the 2 position of acetate, while in MSM they occurred only on carbons derived from the 1 position. These results suggest that the MSM methyltransferase MMAS-1 operates in an inverted manner to that of MTB. PMID- 11437393 TI - Polysaccharide-polynucleotide complexes (IV): antihydrolysis effect of the schizophyllan/poly(C) complex and the complex dissociation induced by amines. AB - Polymer complexes formed by schizophyllan and poly(C) showed a unique antihydrolysis effect when poly(C) was subjected to hydrolysis under basic conditions. The complexation reduced the hydrolysis rate to 80% of the control (i.e., poly(C) itself). However, when we added oligoamines with the intention of catalyzing the hydrolysis, the oligoamines induced dissociation of the complex instead of acceleration of the hydrolysis. PMID- 11437394 TI - Adjuvant high-dose medroxyprogesterone acetate for early breast cancer: 13 years update in a multicentre randomized trial. AB - The authors updated their report on a randomized trial initiated in 1982 comparing, in early breast cancer, high-dose IM Medroxyprogesterone acetate (HD MPA) adjuvant hormonotherapy during 6 months with no hormonotherapy; node positive patients also received 6 courses of IV CMF (day 1, day 8; q.4 weeks). 246 node-negative (NN) and 270 node-positive (NP) patients had been followed for a median duration of 13 years. Previous results were confirmed in this analysis on mature data. In NN patients, relapse-free survival (RFS) was improved in the adjuvant hormonotherapy arm, regardless of age while overall survival (OAS) was also increased in younger (less then 50 years) patients. In the whole group of NP patients, no difference was seen regarding RFS or OAS. However, an age-dependant opposite effect was observed: younger patients (< 50) experienced a worse and significant outcome of relapse-free and overall survivals when receiving adjuvant HD-MPA while older patients (> or = 50) enjoyed a significant improvement of their relapse-free survival. For both NN and NP patients, differences in overall survivals observed in older women with a shorter follow-up, were no longer detected. PMID- 11437395 TI - Fractionated administration of irinotecan and cisplatin for treatment of non small-cell lung cancer: a phase II study of Okayama Lung Cancer Study Group. AB - A phase II study of fractionated administration of irinotecan (CPT-11) and cisplatin (CDDP) in patients with non-small-cell lung cancer (NSCLC) was conducted. Between January 1996 and January 1998, 44 previously untreated patients with stage IIIB or IV NSCLC were enrolled. CDDP at a dose of 60 mg x m( 2) was given first and followed by CPT-11 at a dose of 50 mg x m(-2). Both drugs were given by 1-hour infusion on days 1 and 8, and repeated every 4 weeks up to 4 cycles. 42 patients were evaluated for response and 44 for survival and toxicity. 20 patients (48%: 95% confidence interval 32-63%) achieved an objective response. The median duration of responses was 8 months, and the median survival time and the 1-year survival rate were 12.5 months and 56.8%, respectively. Major toxicities were neutropenia and diarrhoea. Grade 3 or 4 neutropenia occurred in 70.5% of the patients and one patient died of sepsis. Grade 3 or 4 diarrhoea was experienced in 25.0%, but manageable by conventional therapy. In conclusion, fractionated administration of CPT-11 and CDDP was highly effective for advanced NSCLC with manageable toxicities. PMID- 11437396 TI - Aneuploidy and prognosis of non-small-cell lung cancer: a meta-analysis of published data. AB - In lung cancer, DNA content abnormalities have been described as a heterogeneous spectrum of impaired tumour cell DNA histogram patterns. They are merged into the common term of aneuploidy and probably reflect a high genotypic instability. In non-small-cell lung cancer, the negative effect of aneuploidy has been a subject of controversy inasmuch as studies aimed at determining the survival-DNA content relationship have reported conflicting results. We made a meta-analysis of published studies aimed at determining the prognostic effect of aneuploidy in surgically resected non-small-cell lung cancer. 35 trials have been identified in the literature. A comprehensive collection of data has been constructed taking into account the following parameters: quality of specimen, DNA content assessment method, aneuploidy definition, histology and stage grouping, quality of surgical resection and demographic characteristics of the analysed population. Among the 4033 assessable patients, 2626 suffered from non-small-cell lung cancer with aneuploid DNA content (overall frequency of aneuploidy: 0.65; 95% CI: (0.64 0.67)). The DerSimonian and Laird method was used to estimate the size effects and the Peto and Yusuf method was used in order to generate the odds ratios (OR) of reduction in risk of death for patients affected by a nearly diploid (non aneuploid) non-small-cell lung cancer. Survivals following surgical resection, from 1 to 5 years, were chosen as the end-points of our meta-analysis. Patients suffering from a nearly diploid tumour benefited from a significant reduction in risk of death at 1, 2, 3 and 4 years with respective OR: 0.51, 0.51, 0.45 and 0.67 (P< 10(-4)for each end-point). 5 years after resection, the reduction of death was of lesser magnitude: OR: 0.87 (P = 0.08). The test for overall statistical heterogeneity was conventionally significant (P< 0.01) for all 5 end points, however. None of the recorded characteristics of the studies could explain this phenomenon precluding a subset analysis. Therefore, the DerSimonian and Laird method was applied inasmuch as this method allows a correction for heterogeneity. This method demonstrated an increase in survival at 1, 2, 3, 4 and 5 years for patients with diploid tumours with respective size effects of 0.11, 0.15, 0.20, 0.20 and 0.21 (value taking into account the correction for heterogeneity;P< 10(-4)for each end-point). Patients who benefit from a surgical resection for non-small-cell lung cancer with aneuploid DNA content prove to have a higher risk of death. This negative prognostic factor decreases the probability of survival by 11% at one year, a negative effect deteriorating up to 21% at 5 years following surgery. PMID- 11437397 TI - Body surface area estimation in children using weight alone: application in paediatric oncology. AB - The majority of chemotherapy regimens and trials specify doses of cytotoxic drugs normalized to body surface area. Estimation of BSA in paediatric patients is particularly problematic, as conventional nomograms require accurate determination of both height and weight. The chemotherapy standards group of the UKCCSG (United Kingdom Children's Cancer Study Group) has evaluated a method for calculation of body surface area (BSA) estimation, based solely on patient weight. In comparison with BSA estimations using 2 commonly used methods, which require both weight and height measurements, deviation in the estimate of BSA was less than 10%. This method may be extended to the dosing of chemotherapeutic agents in infants of body weight less than 10 kg, with appropriate recommendations for dose modification. Until better correlates of drug clearance, such as GFR for carboplatin, are identified BSA is used to standardize doses for most chemotherapeutic agents. The formula presented here provides a more robust and reliable method of calculation of BSA from weight alone. Although this approach has been shown to be equivalent to other currently used methods, care should be taken extending this calculation of BSA to children less than 10 kg, to obese patients and to those with cachexia. PMID- 11437398 TI - A randomized controlled trial to evaluate the role of interferon as initial and maintenance therapy in patients with follicular lymphoma. AB - The purpose of this study was to evaluate the role of interferon as initial and maintenance therapy in patients with newly diagnosed follicular lymphoma. Between 1984 and 1994, 204 patients with newly diagnosed Stage III or Stage IV follicular lymphoma were randomized to receive either, Chlorambucil (CB): 10 mg daily for 6 weeks, followed by a 2-week interval, with 3 subsequent 2-week treatment periods at the same dose, separated by 2-week intervals, or, CB given concurrently with interferon (IFN). IFN was given at a dose of 3 x 10(6)units thrice weekly, subcutaneously, throughout the 18-week treatment period. Responding patients were subsequently randomized to receive maintenance IFN at the dose and schedule described above, or to expectant management. The overall response rate was 161/204 (78%), complete remission being achieved in 24% of patients. Neither the addition of IFN to the initial treatment, nor the use of maintenance IFN influenced response rate, remission duration or survival. This study was undertaken to determine whether IFN, given in combination with, and then subsequent to, CB would alter the clinical course of patients with follicular lymphoma. Disappointingly, this objective was not achieved, no advantage having been demonstrated for the addition of IFN. PMID- 11437399 TI - CAG and GGC repeat polymorphisms in the androgen receptor gene and breast cancer susceptibility in BRCA1/2 carriers and non-carriers. AB - Variation in the penetrance estimates for BRCA1 and BRCA2 mutations carriers suggests that other genetic polymorphisms may modify the cancer risk in carriers. A previous study has suggested that BRCA1 carriers with longer lengths of the CAG repeat in the androgen receptor (AR) gene are at increased risk of breast cancer (BC). We genotyped 188 BRCA1/2 carriers (122 affected and 66 unaffected with breast cancer), 158 of them of Ashkenazi origin, 166 BC cases without BRCA1/2 mutations and 156 Ashkenazi control individuals aged over 56 for the AR CAG and GGC repeats. In carriers, risk analyses were conducted using a variant of the log rank test, assuming two sets of risk estimates in carriers: penetrance estimates based on the Breast Cancer Linkage Consortium (BCLC) studies of multiple case families, and lower estimates as suggested by population-based studies. We found no association of the CAG and GGC repeats with BC risk in either BRCA1/2 carriers or in the general population. Assuming BRCA1/2 penetrance estimates appropriate to the Ashkenazi population, the estimated RR per repeat adjusted for ethnic group (Ashkenazi and non-Ashkenazi) was 1.05 (95%CI 0.97-1.17) for BC and 1.00 (95%CI 0.83-1.20) for ovarian cancer (OC) for CAG repeats and 0.96 (95%CI 0.80 1.15) and 0.90 (95%CI 0.60-1.22) respectively for GGC repeats. The corresponding RR estimates for the unselected case-control series were 1.00 (95%CI 0.91-1.10) for the CAG and 1.05 (95%CI 0.90-1.22) for the GGC repeats. The estimated relative risk of BC in carriers associated with > or =28 CAG repeats was 1.08 (95%CI 0.45-2.61). Furthermore, no significant association was found if attention was restricted to the Ashkenazi carriers, or only to BRCA1 or BRCA2 carriers. We conclude that, in contrast to previous observations, if there is any effect of the AR repeat length on BRCA1 penetrance, it is likely to be weak. PMID- 11437400 TI - Risk of cancer in patients with dermatomyositis or polymyositis, and follow-up implications: a Scottish population-based cohort study. AB - We conducted a national, retrospective population-based cohort study of 705 patients hospitalized with a first diagnosis of dermatomyositis (DM) or polymyositis (PM) during 1982-1996 based on linkage of hospital discharge, cancer registration, and death records in Scotland. Risks of cancer were assessed by calculating standardized incidence ratios (SIR). A first malignancy was diagnosed concurrently or subsequently in 50 patients with DM (SIR 7.7, 95% CI 5.7-10.1), and 40 patients with PM (2.1, 1.5-2.9). Significantly elevated risks were observed for lung, cervix uteri, and ovarian cancer in patients with DM, and for Hodgkin's disease in patients with PM. The excess risk of cancer was highest around the time of diagnosis, and for patients with DM remained high for at least 2 years. Risks were elevated for both sexes but only significantly so for females, and were highest in patients aged 45-74 years at the time of diagnosis for DM and 15-44 for PM. PMID- 11437401 TI - Risk factors for cancer of the oral cavity and oro-pharynx in Cuba. AB - In terms of worldwide levels, Cuba has an intermediate incidence of cancer of the oral cavity and oro-pharynx. We studied 200 cases of cancer of the oral cavity and pharynx, of whom 57 women (median age = 64) and 200 hospital controls, frequency matched with cases by age and sex, in relation to smoking and drinking history, intake of 25 foods or food groups, indicators of oral hygiene and sexual activity, and history of sexually transmitted diseases. Odds ratios (OR) and 95% confidence intervals (CI) were obtained from unconditional multiple logistic regressions and adjusted for age, sex, area of residence, education, and smoking and drinking habits. In the multivariate model, high educational level and white collar occupation, but not white race, were associated with halving of oral cancer risk. Smoking > or =30 cigarettes per day showed an OR of 20.8 (95% CI: 8.9-48.3), similar to smoking > or =4 cigars daily (OR = 20.5). Drinking > or = 70 alcoholic drinks per week showed an OR of 5.7 (95% CI: 1.8-18.5). Hard liquors were by far the largest source of alcohol. Increased risk was associated with the highest tertile of intake for maize (OR = 1.9), meat (OR = 2.2) and ham and salami (OR = 2.0), whereas high fruit intake was associated with significantly decreased risk (OR = 0.4). Among indicators of dental care, number of missing teeth and poor general oral condition at oral inspection showed ORs of 2.7 and 2.6, respectively. Number of sexual partners, marriages or contacts with prostitutes, practice of oral sex and history of various sexually transmitted diseases, including genital warts, were not associated with oral cancer risk. 82% of oral cancer cases in Cuba were attributable to tobacco smoking, 19% to smoking cigars or pipe only. The fractions attributable to alcohol drinking (7%) and low fruit intake (11%) were more modest. Thus, decreases in cigarette and cigar smoking are at present the key to oral cancer prevention in Cuba. PMID- 11437402 TI - Differential expression and localization of TIMP-1 and TIMP-4 in human gliomas. AB - Studies have suggested that an imbalance of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) may contribute to the malignant phenotype of gliomas. In this study, we have undertaken a detailed analysis of expression of the TIMP family in normal human brain and malignant gliomas at both the mRNA and protein level. Reverse transcription-PCR (RT-PCR) analyses of total RNA from surgical tumour specimens revealed unique expression patterns for the 4 members of the TIMP family, with TIMP-1 and -4 showing positive and negative correlations, respectively, with glioma malignancy. By RT-PCR, TIMP-2 and TIMP-3 expression did not change with tumour grade. In situ hybridization localized TIMP 1 to glial tumour cells and also to the surrounding tumour vasculature. TIMP-4 transcripts were predominantly localized to tumour cells, though minor expression was found in vessels. Recombinant TIMP-4 reduced invasion of U251 glioma cells through Matrigel, and U87 clones overexpressing TIMP-4 showed reduced invasive capacity in vitro. TIMP-4, but not TIMP-1, blocked Membrane Type-1-MMP-mediated progelatinase-A (MMP-2) activation in human umbilical vein endothelial cells. The differential expression and localization of individual TIMPs may contribute to the pathophysiology of human malignant gliomas, particularly with regard to tumour vascularization. PMID- 11437403 TI - Mutations of the beta- and gamma-catenin genes are uncommon in human lung, breast, kidney, cervical and ovarian carcinomas. AB - Beta-catenin forms complexes with Tcf and Lef-1 and functions as a transcriptional activator in the Wnt signalling pathway. Although recent investigations have been focused on the role of the adenomatous polyposis coli (APC)/ beta-catenin/Tcf pathway in human tumorigenesis, there have been very few reports on mutations of the beta-catenin gene in a variety of tumour types. Using PCR and single-strand conformational polymorphism analysis, we examined 93 lung, 9 breast, 6 kidney, 19 cervical and 7 ovarian carcinoma cell lines for mutations in exon 3 of the beta-catenin gene. In addition, we tested these same samples for mutations in the NH2-terminal regulatory region of the gamma-catenin gene. Mutational analysis for the entire coding region of beta-catenin cDNA was also undertaken in 20 lung, 9 breast, 5 kidney and 6 cervical carcinoma cell lines. Deletion of most beta-catenin coding exons was confirmed in line NCI-H28 (lung mesothelioma) and a silent mutation at codon 214 in exon 5 was found in HeLa (cervical adenocarcinoma). A missense mutation at codon 19 and a silent mutation at codon 28 in the NH2-terminal regulatory region of the gamma-catenin gene were found in H1726 (squamous cell lung carcinoma) and H1048 (small cell lung carcinoma), respectively. Neither deletions nor mutations of these genes were detected in the other cell lines examined. These results suggest that beta- and gamma-catenins are infrequent mutational targets during development of human lung, breast, kidney, cervical and ovarian carcinomas. PMID- 11437404 TI - Adenomatous polyposis coli (APC) gene promoter hypermethylation in primary breast cancers. AB - Similar to findings in colorectal cancers, it has been suggested that disruption of the adenomatous polyposis coli (APC)/beta-catenin pathway may be involved in breast carcinogenesis. However, somatic mutations of APC and beta- catenin are infrequently reported in breast cancers, in contrast to findings in colorectal cancers. To further explore the role of the APC/beta-catenin pathway in breast carcinogenesis, we investigated the status of APC gene promoter methylation in primary breast cancers and in their non-cancerous breast tissue counterparts, as well as mutations of the APC and beta- catenin genes. Hypermethylation of the APC promoter CpG island was detected in 18 of 50 (36%) primary breast cancers and in none of 21 non-cancerous breast tissue samples, although no mutations of the APC and beta- catenin were found. No significant associations between APC promoter hypermethylation and patient age, lymph node metastasis, oestrogen and progesterone receptor status, size, stage or histological type of tumour were observed. These results indicate that APC promoter CpG island hypermethylation is a cancer-specific change and may be a more common mechanism of inactivation of this tumour suppressor gene in primary breast cancers than previously suspected. PMID- 11437405 TI - Plasma insulin-like growth factor binding protein-3 proteolysis is increased in primary breast cancer. AB - Fasting blood samples were obtained before definitive surgery or biopsy in 128 patients referred to the department of surgery with suspected or manifest breast cancer. Insulin-like growth factor (IGF)-I, IGF-II and free IGF-I were measured by radioimmunoassay/immunoradiometric assay, while IGFBP-3 proteolysis was evaluated by Western immunoblot. 12 patients had ductal carcinoma in situ benign conditions, while staging revealed metastatic disease in 15 of 16 patients with invasive cancers. IGFBP-3 proteolysis above the normal range was recorded in 19 patients with invasive cancers, but in none of the patients suffering from DCIS/benign conditions. Increased IGFBP-3 proteolysis was most frequently recorded in patients harbouring large tumours and metastatic disease (Stage I: 0/19, 0%; Stage II: 3/45, 7%, Stage III: 9/37, 24%, and Stage IV: 7/15, 47%). IGFBP-3 proteolysis was significantly higher in Stage III (P =0.01) and IV (P< 0.001) patients compared to the other stage groups (P = 0.001). IGF-I and IGF-II correlated negatively to IGFBP-3 proteolysis and age. Plasma levels of IGF-I and II were significantly lower in patients with elevated IGFBP-3 proteolysis compared to those within the normal range. Our findings reveal alterations in the IGF-system among a substantial number of patients with large primary breast cancers. PMID- 11437406 TI - Cellular mechanisms of bone resorption in breast carcinoma. AB - The cellular mechanisms that account for the increase in osteoclast numbers and bone resorption in skeletal breast cancer metastasis are unclear. Osteoclasts are marrow-derived cells which form by fusion of mononuclear phagocyte precursors that circulate in the monocyte fraction. In this study we have determined whether circulating osteoclast precursors are increased in number or have an increased sensitivity to humoral factors for osteoclastogenesis in breast cancer patients with skeletal metastases (+/- hypercalcaemia) compared to patients with primary breast cancer and age-matched normal controls. Monocytes were isolated and cocultured with UMR 106 osteoblastic cells in the presence of 1,25 dihydroxyvitamin D3[1,25(OH)2D3] and human macrophage colony stimulating factor (M-CSF) on coverslips and dentine slices. Limiting dilution experiments showed that there was no increase in the number of circulating osteoclast precursors in breast cancer patients with skeletal metastases (+/- hypercalcaemia) compared to controls. Osteoclast precursors in these patients also did not exhibit increased sensitivity to 1,25(OH)2D3or M-CSF in terms of osteoclast formation. The addition of parathyroid hormone-related protein and interleukin-6 did not increase osteoclast formation. The addition of the supernatant of cultured breast cancer cell lines (MCF-7 and MDA-MB-435), however, significantly increased monocyte osteoclast formation in a dose-dependent fashion. These results indicate that the increase in osteoclast formation in breast cancer is not due to an increase in the number/nature of circulating osteoclast precursors. They also suggest that tumour cells promote osteoclast formation in the bone microenvironment by secreting soluble osteoclastogenic factor(s). PMID- 11437407 TI - Prognostic impact of urokinase-type plasminogen activator receptor (uPAR) in cytosols and pellet extracts derived from primary breast tumours. AB - Using a previously developed enzyme-linked immunosorbent assay (ELISA), the levels of the receptor for urokinase-type plasminogen activator (uPAR) were determined in cytosols and corresponding membrane pellets derived from 878 primary breast tumours. The levels of uPAR in the pellet extracts were more than 3-fold higher than those measured in the cytosols (P< 0.001). Moreover, the uPAR levels in the two types of extracts were weakly, though significantly, correlated with each other (rS= 0.20, P< 0.001). In Cox univariate analysis, high cytosolic levels of uPAR were significantly associated with reduced overall survival (OS) and relapse-free survival (RFS). The levels of uPAR in pellet extracts appeared not to be related with patient survival. In multivariate analysis, elevated levels of uPAR measured in cytosols and pellet extracts were found to be independent predictors of poor OS, not RFS. The prediction of poor prognosis on the basis of high uPAR levels emphasizes its important role in plasmin-mediated degradation of extracellular matrix proteins during cancer invasion and metastasis. PMID- 11437408 TI - Vascular endothelial growth factor-C expression and its relationship to pelvic lymph node status in invasive cervical cancer. AB - Vascular endothelial growth factor-C (VEGF-C) has been implicated in lymphangiogenesis, the process of new lymphatics formation. The present study investigated VEGF-C mRNA expression in invasive cervical cancer tissue. Additionally, the association of VEGF-C mRNA with clinicopathological features was examined. VEGF-C mRNA expression was assessed by reverse transcription polymerase chain reaction using beta-action as an internal control. 75 patients presenting with invasive cervical cancer were included in the trial. VEGF-C mRNA expression was markedly higher in tumours in which pelvic lymph node metastasis was diagnosed by magnetic resonance (MR) imaging (P = 0.002). 53 patients displaying stage Ib-IIb cervical cancer underwent radical hysterectomy and pelvic lymphadenectomy. VEGF-C expression was significantly higher in tumours exhibiting deep stromal invasion, pelvic lymph node metastasis and lymph-vascular space involvement (P = 0.016, P = 0.006 and P = 0.036, respectively). Multivariate analysis revealed VEGF-C mRNA expression to be the sole independent factor influencing pelvic lymph node metastasis. Subjects demonstrating VEGF-C mRNA expression displayed significantly poorer prognoses than those lacking VEGF-C mRNA expression (P = 0.049). These findings provide evidence supporting the involvement of VEGF-C expression in the promotion of lymph node metastasis in cervical cancer. Furthermore, examination of VEGF-C expression in biopsy specimens may be beneficial in the prediction of pelvic lymph node metastasis. PMID- 11437409 TI - Tcf-3 expression and beta-catenin mediated transcriptional activation in aggressive fibromatosis (desmoid tumour). AB - Aggressive fibromatosis harbours mutations resulting in beta-catenin protein stabilization. Primary cell cultures demonstrate constitutive tcf activation in aggressive fibromatosis. Expression and co-immunoprecipitation studies suggest that beta-catenin binds and activates tcf-3 in this tumour. This is the first demonstration of tcf-3 activation by beta-catenin stabilization in a human neoplastic process. PMID- 11437410 TI - Quantitative measurement of thyroglobulin mRNA in peripheral blood of patients after total thyroidectomy. AB - Previous studies have reported the clinical usefulness of reverse transcription polymerase chain reaction (RT-PCR) detection of thyroglobulin (TG) mRNA in the peripheral blood of patients with differentiated thyroid carcinoma. To evaluate this usefulness, we measured TG mRNA in the peripheral blood of patients diagnosed with thyroid carcinoma after total thyroidectomy by real-time quantitative RT-PCR using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA as an internal control. Surprisingly, we detected TG mRNA in all samples obtained after total thyroidectomy, including those from 4 medullary carcinomas. Further, there was no statistical difference in expression levels of TG mRNA in the patients with or without metastasis, and no significant correlation was found between serum TG concentrations and the expression levels of TG mRNA. These results give rise to a question regarding the clinical applications of not only RT-PCR detection but also quantitative measurement of TG mRNA in peripheral blood. PMID- 11437411 TI - High density oligonucleotide array analysis of interferon- alpha2a sensitivity and transcriptional response in melanoma cells. AB - Interferon alpha (IFN-alpha) represents an adjuvant therapy of proven effectiveness in increasing disease-free interval and survival in subgroups of melanoma patients. Since high doses of cytokine are required, the treatment is often accompanied by toxic side effects. Furthermore, naturally occurring insensitivity to IFN-alpha may hamper its therapeutic efficacy. Clinical, molecular or immunological markers enabling the selection of potential responders have not been identified so far. To explore the molecular basis of IFN-alpha responsiveness, we analysed the expression pattern of about 7000 genes in IFN alpha sensitive and resistant cell lines and we compared the transcription profiles of cells cultured in the presence or absence of the cytokine using high density oligonucleotide arrays. Melanoma cell lines were screened for their sensitivity to proliferation inhibition and HLA class I induction upon IFN-alpha treatment by standard 3H-thymidine incorporation and flow-cytometry. The study of 4 sensitive and 2 resistant cell lines allowed the identification of 4 genes (RCC1, IFI16, hox2 and h19) preferentially transcribed in sensitive cells and 2 (SHB and PKC-zeta) preferentially expressed in resistant cells. IFN-alpha stimulation resulted in the expression of a panel of 19 known inducible genes in sensitive but not in resistant cells. Moreover a group of 30 novel IFN-alpha inducible genes was identified. These data may provide a useful basis to develop diagnostic tools to select potential IFN-alpha responders eligible for treatment, while avoiding unnecessary toxicity to non-responders. Furthermore, by extending the knowledge of the polymorphic effects of IFN-alpha on gene expression, they offer novel clues to the study of its pleiotropic toxicity. PMID- 11437412 TI - Induction of apoptosis in prostate carcinoma cells by BH3 peptides which inhibit Bak/Bcl-2 interactions. AB - Interactions between proteins of the Bcl-2 family play an important role in the regulation of apoptosis. Anti-apoptotic family members can heterodimerize with pro-apoptotic family members and antagonize their function, thus protecting against death. In cells protected from death by overexpression of Bcl-2 much of the Bax is present in Bax/Bcl-2 hetero-multimers and its death signal is blocked as it cannot homodimerize. This led us to use the Bcl-2/Bax heterodimer as a target for new compounds which may provide a therapy particularly suited to tumour cells for which resistance to conventional therapy is associated with elevated expression of Bcl-2. We assessed whether apoptosis could be induced in prostate tumour cells by blocking this heterodimerization with synthetic peptide sequences derived from the BH3 domain of pro-apoptotic Bcl-2 family members. Prostate cells were found to undergo up to 40% apoptosis 48 h following the introduction of synthetic peptides from the BH3 domains of Bax and Bak. The caspase inhibitor z-VAD.fmk provided protection against apoptosis mediated by these peptides. Immunoprecipitation studies revealed that introduction of peptides derived from the BH3 regions of Bak and Bax into cells blocked Bak/Bcl-2 heterodimerization. These data suggest that by blocking the dimerization through which Bcl-2 would normally inhibit apoptosis the apoptotic pathway driven by Bak was re-opened. PMID- 11437413 TI - Low-dose retinoic acid enhances in vitro invasiveness of human oral squamous-cell carcinoma cell lines. AB - Retinoids inhibit the proliferation of several types of tumour cells, and are used for patients with several malignant tumours. In this study, we examined the effect of retinoic acids (RAs) on the invasive potentials of the oral squamous cell carcinoma (SCC) cells, BHY and HNt. BHY cells expressed all of retinoid nuclear receptors (RARalpha, beta, gamma, and RXRalpha) and cytoplasmic retinoic acid binding proteins (CRABP1 and CRABP2). HNt cells lacked the expression of RARbeta, but expressed other nuclear receptors and CRABPs. All-trans retinoic acid (ATRA) and 13-cis retinoic acid (13-cisRA) (10(-6)and 10(-7)M) inhibited the growth of the cells, but low-dose ATRA and 13-cisRA (10(-8)M) marginally affected the growth of the cells. Surprisingly, low-dose RAs enhanced the activity of tissue-type plasminogen activator (tPA), and activated pro-matrix metalloproteinases (proMMP2 and proMMP9). Activation of proMMP2 and proMMP9 was inhibited by aprotinin, a serine-proteinase, tPA inhibitor. Furthermore, low-dose RAs enhanced the in vitro invasiveness of BHY cells. These results indicate that low-dose RAs enhances the in vitro invasiveness of oral SCC cells via an activation of proMMP2 and proMMP9 probably mediated by the induction of tPA. PMID- 11437414 TI - Therapy of human non-small-cell lung carcinoma using antibody targeting of a modified superantigen. AB - Superantigens activate T-cells by linking the T-cell receptor to MHC class II on antigen-presenting cells, and novel reactivity can be introduced by fusing the superantigen to a targeting molecule. Thus, an antibody-targeted superantigen, which activates T cells to destroy tumour cells, might be used as cancer therapy. A suitable target is the 5T4 oncofetal antigen, which is expressed on many carcinomas. We constructed a fusion protein from a Fab of a monoclonal antibody recognizing the 5T4 antigen, and an engineered superantigen. The recombinant product 5T4FabV13-SEA(D227A)bound the 5T4 antigen expressed on the human non small-cell lung cancer cell line Calu-1 with a Kd of 1.2 nM while the substitution of Asp227 to Ala in the superantigen moiety reduced binding activity to MHC class II. 5T4FabV13-SEA(D227A)tumour reactivity was demonstrated in 7/7 NSCLC samples by immunohistochemistry, while normal tissue reactivity was low to moderate. 5T4FabV13-SEA(D227A)induced significant T-cell-dependent in vitro killing of sensitive 5T4 bearing Calu-1 cells, with maximum lysis at 10(-10)M, while the capacity to lyse MHC class II expressing cells was approximately 1000 times less effective. Immunotherapy of 5T4FabV13-SEA(D227A)against human NSCLC was investigated in SCID mice reconstituted with human peripheral blood mononuclear cells. Mice carrying intreperitoneally growing Calu-1 cells showed significant reduction in tumour mass and number after intravenous therapy with 5T4FabV13-SEA(D227A). Thus, 5T4FabV13-SEA(D227A)has highly attractive properties for therapy of human NSCLC. PMID- 11437416 TI - Chemotherapy and quality of life in advanced NSCLC. PMID- 11437417 TI - Evaluating new treatments for advanced cancer. PMID- 11437419 TI - Functional reconstruction of mobile tongue and suprahyoid muscles after resection of cancer of the tongue. AB - The mobile tongue and the suprahyoid muscles were reconstructed in a man who had had a total resection of the mobile tongue with the oral floor and bilateral radical neck dissection (extensive resection of the bilateral suprahyoid muscles) for cancer of the tongue. A rectus abdominis musculocutaneous flap with the tenth intercostal nerve anastomosed to the remaining hypoglossal nerve was used for the reconstruction. The rectus abdominis muscle was inserted between the mandible and the hyoid bone, taking account of the anatomical positions of the tongue and suprahyoid muscles; the rectus sheath was then firmly sutured to the mandible and hyoid bone. This procedure created firm tendinous insertions between the mandible and hyoid bone. After the operation, the reconstructed tongue showed no sign of atrophy, and the reconstructed supra-hyoid muscles (rectus abdominis muscle) could be moved, and the hyoid bone could be raised. PMID- 11437420 TI - Changes in masticatory function after orthognathic treatment in patients with mandibular prognathism. AB - Changes in masticatory function were measured in 27 patients in whom mandibular prognathism was corrected surgically. The mean value of masticatory efficiency before treatment was 46% of that of control subjects with normal occlusion. It improved, but remained at 60% of the control value postoperatively. Similar changes were seen in the number and area of occlusal contacts and the integrated muscle activities of the masseter and temporalis on the chewing side, but the postoperative improvement in masticatory efficiency was mainly the result of improvement in masseter activity. The mean values of masticatory cycle variables in the patient group did not differ significantly from those of the controls. Their preoperative mean coefficients of variation, which were significantly higher than those of the controls, decreased significantly postoperatively. These results suggest that the stability of masticatory rhythm was improved by orthognathic surgery. PMID- 11437421 TI - Effect of the angle of the screw on the stability of the mandibular sagittal split ramus osteotomy: a study in sheep mandibles. AB - We compared the biomechanical stability of 60 degrees and 90 degrees angles for insertion of screws for fixation of sagittal split ramus osteotomies (SSROs) in 10 sheep mandibles in vitro. After 5 mm advancements, the osteotomies were fixed with screws 2 mm in diameter inserted with a torque control screwdriver at either 60 degrees or 90 degrees to the long axis of bone, in a matched pair experimental design. All specimens were then loaded in a physiological manner and the load displacement of the osteotomies was recorded. Data for resistance to movement were compared by a paired t -test. The mean (SD) resistance to movement for the 60 degrees and 90 degrees screw insertions was 63.4 (27.7) N/mm and 59 (22.3) N/mm respectively. There was no significant difference between the groups. This correlates with clinical findings. PMID- 11437422 TI - Outcome of extra-oral craniofacial endosseous implants. AB - We report our experience of endosseous implants designed to retain various types of facial prostheses that were inserted by the West of Scotland Oral and Maxillofacial Surgery Service between January 1988 and September 1998. Forty seven patients were given a total of 150 implants, most of which were auricular (n= 85) or orbital (n= 53). The functioning success rates were 100% and 75%, respectively. Soft tissue infection was the commonest complication, and could occur at any time after exposure of the implant, suggesting that life-long follow up is required. PMID- 11437423 TI - Venous obstruction caused by a self-adhesive drape: a cautionary tale. AB - The use of disposable drapes during operations is becoming more widespread, and their advantages are well documented. I describe how use of such a drape led to venous constriction which caused troublesome oozing from the operative sites during sagittal split osteotomy. When the drape was removed, there was a clear red indentation on the patient's neck, and the bleeding stopped immediately. The patient made an uneventful recovery. PMID- 11437424 TI - Median palatine cyst: an unusual presentation of a rare entity. AB - Median palatine cyst is rare. Mostly, it is asymptomatic and usually is discovered incidentally during routine dental or radiological examination. The case that we report has the following unusual features: Firstly, it is the largest cyst to be reported, measuring 5 cm in diameter. Secondly, there was no swelling on the oral surface of the hard palate contrary to other reports. Rather, it caused elevation of the nasal floor and nasal obstruction. Thirdly, it pushed the inferior and caudal end of the septum into the left nasal chamber. The median palatine cyst was surgically removed by a sublabial degloving approach. The cyst was removed in toto and the palatal bone curetted to ensure adequate removal of any nesting cells that could lead to recurrence in the future. PMID- 11437425 TI - Intracranial metastases from oral squamous cell carcinoma. AB - We present two cases of histologically confirmed intracerebral metastases from oral squamous cell carcinoma (SCC). This site of distant spread has not to our knowledge previously been reported. The only common feature in all these cases was the long period over which the patients had untreated primary disease (12--24 months). Both patients developed symptoms from their cerebral deposit within a short time of diagnosis and treatment of the primary disease. They died rapidly of their cerebral metastases despite extensive and effective treatment of the primary site and regional lymph nodes. We recommend that a magnetic resonance (MR) scan of the brain is considered for patients who present with long-standing untreated primary oral SCC. PMID- 11437426 TI - Percutaneous osteosynthesis of the zygomatic buttress. AB - We describe a technique of percutaneous miniplate osteosynthesis of the zygoma, using the transbuccal approach. It can be used in conjunction with an extraoral approach, or in isolation. Excellent access was achieved to the posterior zygomatic buttress and arch of zygoma, and the infraorbital rim. No complications developed in the two cases presented. This technique is a useful addition to the armamentarium of maxillofacial surgeons. PMID- 11437427 TI - Immunohistochemical localization of bone morphogenetic protein-2 in the oral and maxillofacial area of the human embryo. AB - The purpose of this study was to investigate the bone morphogenetic protein-2 (BMP-2) expression in oral and maxillofacial tissue of the human embryo. A human embryo was obtained from a patient with hysterorrhexis. The oral and maxillofacial area of the embryo was analyzed immunohistochemically using specific antibodies to BMP-2. BMP-2 was found mainly localized in developing teeth buds, jaw bone, striated and smooth muscle, spinal cord and nasal tracheal and oesophagal epithelium. However, it was not found in hair follicles. These findings are similar to those of BMP-2 and -4 gene expression in mouse and rat embryos. BMP-2 may be involved in the morphogenesis of oral and maxillofacial tissue in human embryos as well as those of other animals. These results will be of considerable benefit in the restoration and regeneration of the oral and maxillofacial tissue clinically. PMID- 11437428 TI - Experimental osteoinduction by recombinant human bone morphogeneticprotein 2 in tissue with low blood flow: a study in rats. AB - We have compared osteoinduction in soft tissues by 0, 2, 10, and 50 microg of recombinant human bone morphogenetic protein-2 (rhBMP-2) in Wistar rats with a surgically compromised blood supply and controls with a normal blood supply (n = 5 in each group). Three weeks later in the radiographic evaluation, the volume of bone formation in the low-blood-flow groups was similar to that in the controls, and was dependent on the concentration of implanted rhBMP-2. On histological evaluation much woven bone was noted in the low-blood-flow groups, but the volume of bone was less than in the controls. Alkaline phosphatase activity was less and calcium content in the low-blood-flow groups were also less than in the control rats. However, their values increased dose-dependently with implanted rhBMP-2. These results suggest that the osteoinducting activity of rhBMP-2 is reduced in tissues with a low blood supply, but higher doses of rhBMP-2 can be used in compromised tissue. PMID- 11437429 TI - The use of vacuum-formed splints for temporary intermaxillary fixation in the management of unilateral condylar fractures. AB - We report a simple, effective method of managing displaced unilateral condylar fractures with occlusal disruption using vacuum-formed thermoplastic foil splints with bonded wire cleats. The cleats enable intermaxillary fixation in the form of orthodontic elastics to be used, which guide then maintain the occlusion in centric relation. A case is presented in which this technique was used successfully. PMID- 11437430 TI - Unilateral mandibular condylectomy in lambs. AB - The purpose of this experimental study was to investigate the degree of regeneration of the mandibular condyle after unilateral condylectomy in 10-week old lambs. The lambs were killed three months after the operation, and the joints examined radiologically and histologically. Scoring systems were used to assess the radiological changes and histological regeneration of the condyle. All joints showed regeneration of the condylar head postoperatively. The maximum degree of regeneration occurred on the medial side rather than the central or lateral areas. There was a significant correlation among the medial, central, and lateral planes (P<0.05). The reformed articular cartilage was irregular and thin (P<0.01), and the disc was thick in the central plane (P<0.01) compared with the control joints. The temporal bone was normal. This study shows that unilateral condylectomy in the growing period results in some condylar regeneration particularly on the medial side and reformation of some irregular and thin articular cartilage. PMID- 11437431 TI - Matrix metalloproteinase-2 in synovial lavage fluid of patients with disorders of the temporomandibular joint. AB - We examined the matrix metalloproteinase-2 (MMP-2) activity in synovial lavage fluid of patients with disorders of the temporomandibular joint (TMJ) and explored the possible correlationship between MMP-2 activity and radiological changes. We studied 86 patients and 10 healthy volunteers. An arthrogram and a double contrast arthrotomogram were taken to evaluate intra-articular morphological changes. The patients were divided into three groups: no abnormality (n = 36), internal derangement (n = 39), and osteoarthritis (n = 11). Samples of synovial fluid were studied by gelatin zymography, and we sought a correlation between the band detected and radiological findings. ProMMP-2 was detected in all samples and active MMP-2 was detected in 9/36 with no abnormality, 14/39 with internal derangement and 5/11 with osteoarthritis. No active form of MMP-2 was detected in the control group. The incidence of active MMP-2 was high in the internal derangement group and highest in the osteoarthritis group, which suggests that active MMP-2 plays an important part in the development of conditions of the TMJ. PMID- 11437432 TI - Extraneural perineurioma of the face: an unusual cutaneous presentation of an uncommon tumour. AB - Perineurioma is an uncommon, benign tumour of the peripheral nerve sheath that has two major clinicopathological forms -- intraneural and extraneural. We present a case of extraneural perineurioma that occurred at an unusual site -- the facial skin of a 70-year-old woman -- which illustrates the wide, and potentially misleading, clinicopathological spectrum of this poorly recognized tumour. PMID- 11437433 TI - Tuberculous parotitis: report of 3 cases. AB - Tuberculous parotitis, particularly the diffuse form, is rare. I describe here three patients with diffuse tuberculous parotitis, two of whom presented with unilateral parotid swelling while the third had swellings of both glands. All had discharging sinuses. In two of the patients, the parotitis was associated with active pulmonary tuberculosis. In the patient with bilateral parotitis, the disease seemed to be confined to the glands. The diagnosis of tuberculosis was confirmed by the finding of tubercle bacilli in the lesion or the sputum. They all responded well to standard antituberculous drugs. PMID- 11437434 TI - Fixation of a frontozygomatic fracture with a shape-memory staple. AB - A simple method using a staple was successfully used to treat a 74-year-old lady with a fractured frontozygomatic suture. PMID- 11437435 TI - The use of a radial forearm hybrid free flap in the reconstruction of a pharyngeal defect on the opposite side. PMID- 11437436 TI - A tooth in the orbit. PMID- 11437438 TI - Missing links in GSK3 regulation. PMID- 11437439 TI - The MADS-box gene srfA is expressed in a complex pattern under the control of alternative promoters and is essential for different aspects of Dictyostelium development. AB - srfA displays a complex temporal and cell type-specific pattern of expression in Dictyostelium and is expressed by most of its cell types at some stage of their development. This complexity is achieved by the use of alternative promoters. The promoter activity of the proximal region was found to be restricted to a subset of prestalk cells. Little or no associated expression was observed in the lower cup and basal disc during culmination. The middle promoter region was preferentially active in prestalk cells under usual conditions of filter development. Interestingly, during slug migration, the activity of this promoter in posterior prespore cells was strongly induced. The distal region displayed a dual pattern of expression. Thus, before culmination, this region drove lacZ expression in a few cells scattered along the entire structure. However, intense lacZ staining was found in the spores by the end of culmination. We have previously reported that srfA expression is essential for spore differentiation (R. Escalante and L. Sastre, Development 125, 3801-3808). Our novel finding of the expression of the gene in prestalk cells before culmination suggested that it might play additional roles in Dictyostelium development. The study of knockout strains revealed that srfA is also required for proper slug migration. Spore differentiation and slug migration defects were rescued by reexpression of srfA in the null mutant background, under the appropriate promoter control. The expression of srfA under the activity of the distal promoter region was able to rescue spore differentiation but not slug migration. Conversely, reexpression under the control of the middle promoter rescued slug morphogenesis and migration. Our results demonstrate that the correct spatial and temporal pattern of expression of srfA is essential for the different functions that this transcription factor plays in development. PMID- 11437440 TI - Evidence that protein kinase C (PKC) participates in the meiosis I to meiosis II transition in mouse oocytes. AB - Oocytes from LTXBO mice exhibit a delayed entry into anaphase I and frequently enter interphase after the first meiotic division. This unique oocyte model was used to test the hypothesis that protein kinase C (PKC) may regulate the meiosis I-to-meiosis II transition. PKC activity was detected in LTXBO oocytes at prophase I and increased with meiotic maturation, with the highest (P < 0.05) activity observed at late metaphase I (MI). Treatment of late MI-stage oocytes with the PKC inhibitor, bisindolylmaleimide I (BIM), transiently reduced (P < 0.05) M-phase-promoting factor (MPF) activity and promoted (P < 0.05) progression to metaphase II (MII), while mitogen-activated protein kinase (MAPK) activity remained elevated during the MI-to-MII transition. Confocal microscopy analysis of LTXBO oocytes during this transition showed PKC-delta associated with the meiotic spindle and then with the chromosomes at MII. Inhibition of PKC activity also prevented untimely entry into interphase, but only when PKC activity was reduced in oocytes before the progression to MII and thus indicates that the transition into interphase is directly associated with the delayed triggering of anaphase I. Moreover, the defect(s) that initiate activation occur upstream of MAPK, as suppression of PKC activity failed to prevent activation by Mos(tm1Ev)/ Mos(tm1Ev) LTXBO oocytes expressing no detectable MAPK activity. In summary, PKC participates in the regulatory mechanisms that delay entry into anaphase I in LTXBO oocytes, and the disruption promotes untimely entry into interphase. Thus, loss of regulatory control over PKC activity during oocyte maturation disrupts the critical MI-to-MII transition, leading to a precocious exit from meiosis. PMID- 11437441 TI - Loss of Xist imprinting in diploid parthenogenetic preimplantation embryos. AB - We have analysed Xist expression patterns in parthenogenetic and control fertilised preimplantation embryos by using RNA FISH. In normal XX embryos, maternally derived Xist alleles are repressed throughout preimplantation development. Paternal alleles are expressed as early as the 2-cell stage. In parthenogenetic embryos, we observed Xist RNA expression and accumulation from the morula stage onwards, indicating loss of maternal imprinting. In the majority of cells, expression was from a single allele, indicating that X chromosome counting occurs to establish appropriate monoallelic Xist expression. We discuss these data in the context of models for regulation of imprinted and random X inactivation. PMID- 11437442 TI - Zebrafish fgf3 and fgf8 encode redundant functions required for otic placode induction. AB - Members of the fibroblast growth factor (FGF) family of peptide ligands have been implicated in otic placode induction in several vertebrate species. Here, we have functionally analyzed the roles of fgf3 and fgf8 in zebrafish otic development. The role of fgf8 was assessed by analyzing acerebellar (ace) mutants. fgf3 function was disrupted by injecting embryos with antisense morpholino oligomers (MO) specifically designed to block translation of fgf3 transcripts. Disruption of either fgf3 or fgf8 causes moderate reduction in the size of the otic vesicle. Injection of fgf3-MO into ace/ace mutants causes much more severe reduction or complete loss of otic tissue. Moreover, preplacode cells fail to express pax8 and pax2.1, indicating disruption of early stages of otic induction in fgf3-depleted ace/ace mutants. Both fgf3 and fgf8 are normally expressed in the germring by 50% epiboly and are induced in the primordium of rhombomere 4 by 80% epibloy. In addition, fgf3 is expressed during the latter half of gastrulation in the prechordal plate and paraxial cephalic mesendoderm, tissues that either pass beneath or persist near the prospective otic ectoderm. Conditions that alter the pattern of expression of fgf3 and/or fgf8 cause corresponding changes in otic induction. Loss of maternal and zygotic one-eyed pinhead (oep) does not alter expression of fgf3 or fgf8 in the hindbrain, but ablates mesendodermal sources of fgf signaling and delays otic induction by several hours. Conversely, treatment of wild-type embryos with retinoic acid greatly expands the periotic domains of expression of fgf3, fgf8, and pax8 and leads to formation of supernumerary and ectopic otic vesicles. These data support the hypothesis that fgf3 and fgf8 cooperate during the latter half of gastrulation to induce differentiation of otic placodes. PMID- 11437443 TI - Ectopic expression of the Arabidopsis AtMYB23 gene induces differentiation of trichome cells. AB - The control of epidermal cell fate is a complex molecular process and requires the regulatory activity of different transcription factors. Here, we describe the isolation of a member of the Arabidopsis MYB transcription factor family, AtMYB23, that is involved in trichome development. Expression of the AtMYB23 gene under the control of the viral CaMV 35S promoter causes the development of ectopic trichomes. The formation of ectopic trichomes depends on TRANSPARENT TESTA GLABRA1 but not on GLABRA1. The absence of the negative regulator TRIPTYCHON leads to branching of the ectopic trichomes on cotyledons and the formation of ectopic trichomes in the leaf subepidermal cell layer. The CaMV 35S promoter-controlled expression of AtMYB23 can partially rescue the glabra1 mutant phenotype. Together, the presented data indicate that the AtMYB23 gene has partially overlapping functions with GLABRA1 in controlling the initiation of trichome development. PMID- 11437444 TI - The casein kinase I family: roles in morphogenesis. AB - Wnt signals play important roles in development and oncogenesis and are transduced through at least two pathways: a canonical beta-catenin-dependent and a beta-catenin-independent cascade. Casein kinase I (CKI) is required in both invertebrates and vertebrates to transduce canonical Wnt signals. However, its role in the beta-catenin-independent pathway was unknown. During vertebrate embryogenesis, the beta-catenin-independent cascade is thought to control cell movements and has been postulated to be analogous to the Drosophila planar cell polarity pathway, which signals through the JNK cascade. Here, we report that blocking CKI function inhibits embryonic morphogenesis and activates JNK in cell lines. These studies suggest that CKI might also act in the beta-catenin independent pathway and indicate a role for CKI during convergence extension in early vertebrate development. PMID- 11437445 TI - The casein kinase I family in Wnt signaling. AB - The canonical Wnt-signaling pathway is critical for many aspects of development, and mutations in components of the Wnt pathway are carcinogenic. Recently, sufficiency tests identified casein kinase Iepsilon (CKIepsilon) as a positive component of the canonical Wnt/beta-catenin pathway, and necessity tests showed that CKIepsilon is required in vertebrates to transduce Wnt signals. In addition to CKIepsilon, the CKI family includes several other isoforms (alpha, beta, gamma, and delta) and their role in Wnt sufficiency tests had not yet been clarified. However, in Caenorhabditis elegans studies, loss-of-function of a CKI isoform most similar to alpha produced the mom phenotype, indicative of loss-of Wnt signaling. In this report, we examine the ability of the various CKI isoforms to activate Wnt signaling and find that all the wild-type CKI isoforms do so. Dishevelled (Dsh), another positive component of the Wnt pathway, becomes phosphorylated in response to Wnt signals. All the CKI isoforms, with the exception of gamma, increase the phosphorylation of Dsh in vivo. In addition, CKI directly phosphorylates Dsh in vitro. Finally, we find that CKI is required in vivo for the Wnt-dependent phosphorylation of Dsh. These studies advance our understanding of the mechanism of Wnt action and suggest that more than one CKI isoform is capable of transducing Wnt signals in vivo. PMID- 11437446 TI - Neurons and glia in the midline of the higher crustacean Orchestia cavimana are generated via an invariant cell lineage that comprises a median neuroblast and glial progenitors. AB - Midline cells are a common feature of both insects and crustaceans. Midline cells in the insects Schistocerca americana and Drosophila melanogaster have been shown to give rise to pairs of either neurons or glial cells (midline precursor) as well as to repeatedly generate neurons (median neuroblast) or both neurons and glia (median neuroglioblast). This study addresses midline cell lineages in a higher crustacean, the amphipod Orchestia cavimana. In vivo labeling of single midline cells shows that the resulting cell lineage is invariant and that these cells act as progenitors for sets of three glial precursors and one median neuroblast. The progeny are restricted to parasegmental units. The glial precursors give rise to three pairs of glial cells; two of them enwrap the commissures. The median neuroblast gives rise to about 10 cells that differentiate into 3 classes of neurons. The presence of median neuroblasts is also shown for another higher crustacean, the isopod Porcellio scaber using BrdU labeling. This is the first study to analyze the cell lineage of crustacean neurons generated by early ectodermal precursors. A comparison with those of insects demonstrates both conservation and change during the evolution of arthropods. PMID- 11437447 TI - The Caenorhabditis elegans gene lin-26 can trigger epithelial differentiation without conferring tissue specificity. AB - How epithelial cell fates become specified is poorly understood. We have previously shown that the putative C2H2 zinc-finger transcription factor LIN-26 is required for the differentiation of ectodermal and mesodermal epithelial cells in Caenorhabditis elegans. Here, we report that ectopic LIN-26 expression during early gastrulation transforms most blastomeres into epithelial-like cells. Specifically, LIN-26 induced the expression of three epithelial markers: the adherens junction protein JAM-1; DLG-1, which is essential for the assembly of JAM-1 at junctions; and CHE-14, which is involved in apical trafficking. Furthermore, ultrastructural studies revealed that ectopic LIN-26 expression induced the formation of adherens-like junctions. However, ectopic lin-26 expression did not confer any tissue-specific cell fate, such as the epidermal cell fate, as evidenced from the observation that several epidermal-specific genes were not induced. Conversely, we show that epidermal cells displayed some polarity defects in lin-26 mutants. We conclude that lin-26 can induce epithelial differentiation and that epitheliogenesis is not a default pathway in C. elegans. PMID- 11437448 TI - Retinoic acid promotes differentiation of trophoblast stem cells to a giant cell fate. AB - Trophoblast stem cell (TS cell) lines have the ability to differentiate into trophoblast subtypes in vitro and contribute to the formation of placenta in chimeras. In order to investigate the possible role of retinoic acid (RA) in placentation, we analyzed the effects of exogenous RA on TS cells in vitro and the developing ectoplacental cone in vivo. TS cells expressed all subtypes of the retinoid receptor family, with the exception of RARbeta, whose expression was stimulated in response to RA. TS cells treated with RA were compromised in their ability to proliferate and exhibited properties of differentiation into trophoblast giant cells. During TS cell differentiation into trophoblast subtypes induced by withdrawal of FGF4, RA treatment further illustrated its role in the specification of cell fate by the promotion of differentiation into giant cells and the suppression of spongiotrophoblast formation. Moreover, administration of RA during pregnancy resulted in the overabundance of giant cells at the expense of spongiotrophoblast cells. RA hereby acts as an extracellular signal whose potential function can be linked to specification events mediating trophoblast cell fate. Taken together with the spatial patterns of giant-cell formation and RA synthesis in vivo, these findings implicate a function for RA in giant-cell formation during placentation. PMID- 11437449 TI - Regulation of dally, an integral membrane proteoglycan, and its function during adult sensory organ formation of Drosophila. AB - In Drosophila, imaginal wing discs, Wg and Dpp, play important roles in the development of sensory organs. These secreted growth factors govern the positions of sensory bristles by regulating the expression of achaete-scute (ac-sc), genes affecting neuronal precursor cell identity. Earlier studies have shown that Dally, an integral membrane, heparan sulfate-modified proteoglycan, affects both Wg and Dpp signaling in a tissue-specific manner. Here, we show that dally is required for the development of specific chemosensory and mechanosensory organs in the wing and notum. dally enhancer trap is expressed at the anteroposterior and dorsoventral boundaries of the wing pouch, under the control of hh and wg, respectively. dally affects the specification of proneural clusters for dally sensitive bristles and shows genetic interactions with either wg or dpp signaling components for distinct sensory bristles. These findings suggest that dally can differentially regulate Wg- or Dpp-directed patterning during sensory organ assembly. We have also determined that, for pSA, a bristle on the lateral notum, dally shows genetic interactions with iroquois complex (IRO-C), a gene complex affecting ac-sc expression. Consistent with this interaction, dally mutants show markedly reduced expression of an iro::lacZ reporter. These findings establish dally as an important regulator of sensory organ formation via Wg- and Dpp mediated specification of proneural clusters. PMID- 11437450 TI - Bmp6 and Bmp7 are required for cushion formation and septation in the developing mouse heart. AB - The mature heart valves and septa are derived from the cardiac cushions which initially form as local outgrowths of mesenchymal cells within the outflow tract and atrioventricular regions. Endocardial cells respond to signals from the overlying myocardium and undergo an epithelial-to-mesenchymal transformation to invade the intervening extracellular matrix. The molecules that can induce and maintain these cell populations are not known, but many candidates, including several TGFbetas and BMPs, have been proposed based on their expression patterns and activities in other systems. In the present study, we describe the expression of Bmp6 and Bmp7 in overlapping and adjacent sites, including the cardiac cushions during mouse embryonic development. Previous analyses demonstrate that neither of these BMPs is required during cardiogenesis, but analysis of Bmp6;Bmp7 double mutants uncovers a marked delay in the formation of the outflow tract endocardial cushions. A proportion of Bmp6;Bmp7 mutants also display defects in valve morphogenesis and chamber septation, and the embryos die between 10.5 and 15.5 dpc due to cardiac insufficiency. These data provide the first genetic evidence that BMPs are involved in the formation of the cardiac cushions. PMID- 11437451 TI - Wingless capture by Frizzled and Frizzled2 in Drosophila embryos. AB - A variety of factors could influence how far developmental signals spread. For example, the Patched receptor limits the range of its ligand Hedgehog. Somehow, the Frizzled2 receptor has the opposite effect on its ligand. Increasing the level of Frizzled2 stabilizes Wingless and thus extends the Wingless gradient in Drosophila wing imaginal disks. Here we ask whether Frizzled or Frizzled2 affects the spread of Wingless in Drosophila embryos. We show that in the embryonic epidermis, the combined expression of both receptors is lowest in the engrailed domain. This is because expression of Frizzled is repressed by the Engrailed transcription factor, whereas that of Frizzled2 is repressed by Wingless signaling. Receptor downregulation correlates with an early asymmetry in Wingless distribution, characterized by the loss of Wingless staining in the engrailed domain. Raising the expression of either Frizzled or Frizzled2 in this domain prevents the early disappearance of Wingless-containing vesicles. Apparently, Wingless is captured, stabilized, and quickly internalized by either receptor. As far as we can tell, captured Wingless is not passed on to further cells and does not contribute to the spread of Wingless. Receptor downregulation in the posterior compartment may contribute to dampening the signal at the time when cuticular fates are specified. PMID- 11437452 TI - A polychaete hunchback ortholog. AB - We report the first characterization of a segmentation gene homologue in the basal polychaete Capitella capitata using a pan-annelid cross-species antibody to the hunchback-like gene product. In flies, the gap segmentation gene hunchback (hb) encodes a C(2)H(2) zinc-finger transcription factor that plays a pivotal role in patterning the anterior region of the fly body plan. The hb orthologue in Capitella (Cc-hb) is expressed maternally and in all micromere and macromere cells throughout cleavage. At gastrulation, nuclear Cc-hb protein is expressed in the micromere-derived surface epithelium that undergoes epiboly and in the large vegetal blastomeres that gradually become internalized. During organogenesis, Cc hb is expressed in the developing gut epithelium, the prostomial and pygidial epithelium, and in a subset of differentiated neurons in the adult central nervous system. Cc-hb is not expressed in the segmental precursor cells in the trunk. The Cc-hb expression domains in Capitella are similar to those reported for the leech hb orthologue (LZF2), and many of the observed differences between the annelid classes correlate with changes in life history. The lack of detectable annelid hb protein in the trunk at the time of AP pattern formation in leech and in polychaete suggests that the anterior organizing function of hb in flies originated in the arthropod or insect lineage. PMID- 11437453 TI - Defects in pulmonary vasculature and perinatal lung hemorrhage in mice heterozygous null for the Forkhead Box f1 transcription factor. AB - Decreased pulmonary expression of Forkhead Box f1 (Foxf1) transcription factor was associated with lethal alveolar hemorrhage in 55% of the Foxf1 +/- newborn mice. The severity of the pulmonary abnormalities correlates with the levels of Foxf1 mRNA. Defects in alveolarization and vasculogenesis were observed in subsets of the Foxf1 +/- mice with relatively low levels of expression from the normal Foxf1 allele. Lung hemorrhage was coincident with disruption of the mesenchymal-epithelial cell interfaces in the alveolar and bronchiolar regions of the lung parenchyma and was associated with increased apoptosis and reduced surfactant protein B (SP-B) expression. Finally, the lung defect associated with the Foxf1 +/- mutation was accompanied by reduced expression of vascular endothelial growth factor (VEGF), the VEGF receptor 2 (Flk-1), bone morphogenetic protein 4 (Bmp-4), and the transcription factors of the Brachyury T-Box family (Tbx2-Tbx5) and Lung Kruppel-like Factor. Reduction in the level of Foxf1 caused neonatal pulmonary hemorrhage and abnormalities in alveologenesis, implicating this transcription factor in the regulation of mesenchyme-epithelial interaction critical for lung morphogenesis. PMID- 11437454 TI - Novel cell lines promote the discovery of genes involved in early heart development. AB - Clonal cell lines representing early cardiomyocytes would provide valuable reagents for the dissection of the genetic program of early cardiogenesis. Here we describe the establishment and characterization of cell lines from the hearts of transgenic mice and embryos with SV40 large T antigen expressed in the heart forming region. Ultrastructure analysis by transmission electron microscopy showed the primitive, precontractile nature of the resulting cells, with the absence of myofilaments, Z lines, and intercalated disks. Immunohistochemistry, RT-PCR, Northern blots, and oligonucleotide microarrays were used to determine the expression levels of thousands of genes in the 1H and ECL-2 cell lines. The resulting gene-expression profiles showed the transcription of early cardiomyocyte genes such as Nkx2.5, GATA4, Tbx5, dHAND, cardiac troponin C, and SM22-alpha. Furthermore, many genes not previously implicated in early cardiac development were expressed. Two of these genes, Hic-5, a possible negative regulator of muscle differentiation, and the transcription enhancing factor TEF-5 were selected and shown by in situ hybridizations to be expressed in the early developing heart. The results show that the 1H and ECL-2 cell lines can be used to discover novel genes expressed in the early cardiomyocyte. PMID- 11437455 TI - Synaptotagmin VI participates in the acrosome reaction of human spermatozoa. AB - Acrosomal exocytosis is a calcium-dependent secretion event causing the release of the acrosomal contents and the loss of the membranes surrounding the acrosome. The synaptotagmins are a family of calcium-binding proteins that participate in the exocytosis of synaptic vesicles. The ubiquitous synaptotagmin VI isoform was found in human sperm cells by Western blot analysis. Immunocytochemistry at the optical and electron microscopy levels localized the protein to the outer acrosomal membrane. Calcium-triggered acrosomal exocytosis in permeabilized sperm cells was abrogated by a specific anti-synaptotagmin VI antibody, indicating that the protein is required for the process. Moreover, a recombinant fusion protein between glutathione S-transferase and the two calcium and phospholipid binding domains of synaptotagmin VI completely inhibited calcium-triggered exocytosis. Interestingly, phorbol ester-dependent in vitro phosphorylation of this recombinant protein abolished its inhibitory effect. We previously showed that, in permeabilized spermatozoa, addition of active Rab3A triggers acrosomal exocytosis at very low calcium concentration. Rab3A-promoted exocytosis was inhibited by the cytosolic domain of synaptotagmin VI and by the anti synaptotagmin VI antibody, indicating that synaptotagmin is also necessary for Rab-mediated acrosomal content release. In conclusion, the results strongly indicate that synaptotagmin VI is a key component of the secretory machinery involved in acrosomal exocytosis. PMID- 11437457 TI - Evolution of the management and prevention of childhood lead poisoning: dependence of advances in public health on technological advances in the determination of lead and related biochemical indicators of its toxicity. PMID- 11437458 TI - Environmental risk factors and Parkinson's disease: a metaanalysis. AB - The study aim was to examine the association between Parkinson's disease (PD) and exposure to environmental factors such as living in a rural area, well water use, farming, exposure to farm animals, or living on a farm, and pesticides. A series of metaanalyses of peer-reviewed studies were performed, using 16 studies for living in rural area, 18 studies for well water drinking, 11 studies for farming, and 14 studies for pesticides. Prior to the metaanalyses, all studies were reviewed and evaluated for heterogeneity and publication bias. Significant heterogeneity among studies was detected and combined odds ratio (OR) was calculated using the random and the fixed-effect models. The majority of the studies reported consistent elevation in the risk of PD with exposure to environmental factors such as rural living and farming. The combined OR for rural residence was 1.56 [95% confidence interval (95% CI) 1.18-2.07] for all the studies, and 2.17(95% CI 1.54-3.06) for studies performed in United States. The combined OR for well water use was 1.26 (95% CI 0.97-1.64) for all the studies, and 1.44(95% CI 0.92-2.24) for studies done in United States. The combined OR for farming, exposure to farm animals, or living on a farm was 1.42 (95% CI 1.05 1.91) for all studies, and 1.72(95% CI 1.20-2.46) for studies done in United States. The combined OR for pesticides exposure was 1.85(95% CI 1.31-2.60) for all studies, and 2.16(95% CI 1.95-2.39) for studies done in United States. Dose response relationships could not be established due to the imprecise nature of the reported data. Our findings suggest that living in a rural area, drinking well water, farming, and exposure to pesticides may be a risk factor for developing PD. PMID- 11437459 TI - Changes in serum PBB and PCB levels over time among women of varying ages at exposure. AB - The identification of host factors that are predictors of changes in serum polyhalogenated biphenyl contaminants over time has been a difficult challenge in epidemiologic studies of exposed individuals. Of particular concern are age at exposure, reproductive and lactational histories, and changes in body mass index. Using both cross-sectional and longitudinal approaches, this study examined factors related to high initial serum PBB and PCB levels and changes in these levels over time among women of varying ages at exposure (n=1772; age range<1 to 45 years). In 1973, PBB exposure occurred through consumption of farm products contaminated with PBB added to cattle feed. Exposures to PCBs began in 1941 through PCB-contaminated silo sealant deteriorating into animal feed. The Michigan Department of Public Health began enrolling participants in 1977 and has continued to follow them through annual updates. At enrollment, questionnaires were administered to obtain demographic, lifestyle, and anthropometric measurements, medical/reproductive and occupational histories, and contaminated food consumption patterns. Blood samples were collected for PBB and PCB analysis at enrollment for all participants; additional serum tests were done on a subset of the population during follow-up. Median serum levels at enrollment were 2.0 ppb PBB and 5.0 ppb PCB. A decline in serum PBB level over an interval that ranged from 1 to 146 months (median=31) was observed for 44.6% of the women (median=1.0 ppb), while 12.2% showed an increase (median=1.0 ppb). PCB levels declined in 50.3% of the women (median=3.0 ppb) while 12.2% increased (median=2.0 ppb). Relative to women whose contaminant levels were stable, higher initial serum level was a predictor of decline for both PBB and PCB (OR=1.66, 95% CI 1.52 1.82; OR=3.26, 95% CI 2.58-4.12, respectively); a yearly increase in interval between tests was related to declining PCBs (OR=1.65, 95% CI 1.46-1.87). In addition, age< or =10 years at exposure (OR=1.72, 95% CI 1.03-2.86) and residence on a quarantined farm (OR=1.40, 95- CI 1.03-1.90) were predictors of a decrease in PBBs. Factors related to an increase in PBB levels were age< or =10 years at exposure (OR=0.30, 95% CI 0.10-0.96) and initial PBB level (OR=1.24, 95% CI 1.15 1.33); and for PCBs, high initial level (OR=1.34, 95% CI 1.17-1.53) and body mass index (OR=1.07, 95% CI 1.01-1.13). One or more live births during the interval between tests were not related to changing levels of either contaminant; breastfeeding data were not available for examination. Early age at exposure appears to be an important predictor of changes in serum PBB levels over time. PMID- 11437460 TI - Lifestyles, diets, and Native American exposure factors related to possible lead exposures and toxicity. AB - Lead exposure is still a national concern, and it is possible that Native Americans who live on reservations and pursue traditional lifestyles may be at higher risk through both their unique exposure profiles and their potentially greater sensitivity. A major component of the exposure assessment is the diet. For tribal members, traditional lifestyles that include native foods, medicines, and traditional practices have evolved and proven to be the most healthful over many thousands of years of coexistence with the environment. However, a completely traditional diet may not be fully available for a variety of reasons; so, one must also consider the adverse health consequences caused by the loss of healthy native foods and medicines, the contamination of remaining native foods, the inability to practice one's religion, and the possibly lower quality of the substitute diet. Health evaluations of lead exposure on reservations should therefore consider at least two types of diets in addition to the typical suburban diet: (a) traditional diets composed of native foods and medicines that would result in increased exposure if the plants and animals are contaminated and (b) disadvantaged or commodity food diets that result in widespread vitamin and mineral deficits of the sort known to increase absorption of and response to lead. Additional exposure to lead might come from reservation housing which is often older, although the prevalence of lead-based paint on reservations is unknown. The degree of physiological response could also be affected by widespread exposures to other neurotoxins (such as mercury and PCBs in fish), underlying disease patterns, and genetics. Although each of these factors is plausible, their prevalence singly or in combination is unknown. Any correlation between these risk factors and blood lead levels on reservations is also unknown. This paper begins to address these gaps by discussing the range of traditional and current diets that may exist among tribes and methods for developing a whole lifestyle exposure scenario that is appropriate for an individual tribe. Some of the factors discussed in this paper may not apply to the large population of Native Americans who live in urban situations. PMID- 11437461 TI - Evaluation of the HUD lead hazard control grant program: early overall findings. AB - This study evaluates the effectiveness of lead hazard control methods used in the Lead Hazard Control (LHC) grant program of U.S. Department of Housing and Urban Development. The LHC Program awards funds to local jurisdictions to address lead hazards in privately owned, low-income dwellings. Grantees in 14 cities, states, or counties collected environmental data in over 2600-treated dwellings making this the largest study of residential lead hazard control ever undertaken. Grantees employed a range of treatments, the most common being replacement of windows and repair of deteriorated lead-based paint. In this paper, dust lead loading levels and blood lead levels of children (6 months-6 years, if present) were observed at four periods of time (preintervention, immediate, and 6- and 12 months postintervention) in 1212 dwellings. Dust lead loading levels were also observed in a subset of these dwellings at 24- and 36-months postintervention. The geometric mean floor and window dust lead loadings declined at least 50 and 88% (P<0.0001), respectively, immediately postintervention. Three years later, floor dust lead loadings remained at or below the immediate postintervention levels. Window dust lead loadings had moderate increases, but remained substantially reduced from preintervention levels and below clearance standards. At 1 year after intervention, geometric mean age-adjusted blood lead levels had declined from 11.0 to 8.2 microg/dL, a 26% decline (P<0.0001). The LHC Program interventions produced blood lead declines similar to or greater than the percentage changes reported in earlier 1-year lead intervention studies. PMID- 11437462 TI - Metal levels in southern leopard frogs from the Savannah River Site: location and body compartment effects. AB - Tadpoles have been proposed as useful bioindicators of environmental contamination; yet, recently it has been shown that metal levels vary in different body compartments of tadpoles. Metals levels are higher in the digestive tract of bullfrog (Rana catesbeiana) tadpoles, which is usually not removed during such analysis. In this paper we examine the heavy metal levels in southern leopard frog (R. utricularia) tadpoles from several wetlands at the Savannah River Site and test the null hypotheses that (1) there are no differences in metal levels in different body compartments of the tadpoles, including the digestive tract; (2) there are no differences in heavy metal levels among different wetlands; and (3) there are no differences in the ratio of metals in the tail/body and in the digestive tract/body as a function of metal or developmental stage as indicated by body weight. Variations in heavy metal levels were explained by wetland and body compartment for all metals and by tadpole weight for selenium and manganese. In all cases, levels of metals were higher in the digestive tract than in the body or tail of tadpoles. Metal levels were highest in a wetland that had been remediated and lowest in a wetland that was never a pasture or remediated (i.e., was truly undisturbed). Although tadpoles are sometimes eaten by fish and other aquatic predators, leopard frogs usually avoid laying their eggs in ponds with such predators. However, avian predators will eat them. These data suggest that tadpoles can be used as bioindicators of differences in metal levels among wetlands and as indicators of potential exposure for higher-trophic-level organisms, but that to assess effects on the tadpoles themselves, digestive tracts should be removed before analysis. PMID- 11437463 TI - Impact of copper on biomonitoring enzyme ethoxyresorufin-o-deethylase in cultured catfish hepatocytes. AB - The enzyme ethoxyresorufin-o-deethylase (EROD) of the cytochrome P4501A family (CYP1A) in fish liver is increasingly being used as a molecular marker for qualitative and quantitative estimation of aquatic pollution throughout the world. The regulation and expression of this enzyme protein is very important from the toxicological point of view. The regulation of gene expression for this enzyme is mediated by the aromatic hydrocarbon receptor. In addition, cellular glutathione status influences expression of CYP1A. In this study, we explored the relationships among glutathione, EROD, and copper in cultured hepatocytes from Indian catfish. EROD activity in cultured hepatocytes was induced by carbofuran (CF), a widely used agricultural pesticide, and by beta-napthoflavone (BNF), a known inducer of CYP1A. Addition of copper into the culture media of hepatocytes inhibited EROD activity significantly. The activity of EROD elevated by CF and BNF was inhibited in hepatocytes pretreated with CF and BNF exposed to CuSO4. This effect was reflected in the glutathione status of the cells. The level of glutathione was increased by 3.4 and 3.0 times in hepatocytes treated with CF and BNF, respectively. These levels were inhibited in hepatocytes exposed to CuSO(4). Thus, copper interactions with glutathione may play a role in regulating EROD in hepatocytes. PMID- 11437464 TI - DDT environmental persistence from its use in a vector control program: a case study. AB - DDT contamination was investigated in soil, sediment, and chicken eggs from an endemic leishmaniasis area located in Rio de Janeiro City, Brazil. The last DDT application in this area was in 1990, for sand-fly vector control. Sampling campaigns were conducted in 1997 and 1999. DDT was extracted by use of a modified soxhlet apparatus and analysis was performed by gas chromatography with electron capture detector. The results show that, in 1997, soil samples contained up to 351 microg x kg(-1) x d x w. of summation operator DDT near the insecticide sprayed sites. In 1999, the soil concentration decreased to 112 microg x kg(-1) x d x w. of summation operator DDT. Sediments from small creeks also showed low concentrations (up to 32.9 microg x kg(-1) x d x w. of summation operator DDT). Chicken eggs had, on average, 1.98 mg x kg(-1) summation operator DDT (twice FAO's maximum residue limit), comprising 82% of p,p'-DDE. Taking into account the egg results, DDT bioaccumulation is a question of concern. Considering just the egg consumption, it was estimated that DDT intake in the study area is 0.38x10( 4) mg x kg(-1) body weight x day(-1) whereas the reference maximum dose (US EPA) is 5x10(-4) mg x kg(-1) body weight x day(-1). This approach can be used to estimate DDT exposure in other places where DDT contamination may be of concern, especially in places where locally produced animals and eggs are a significant portion of the diet. PMID- 11437465 TI - Mercury contamination in freshwater, estuarine, and marine fishes in relation to small-scale gold mining in Suriname, South America. AB - The extent of mercury contamination in Surinamese food fishes due to small-scale gold mining was investigated by determination of the total mercury concentration in 318 freshwater fishes, 109 estuarine fishes, and 110 fishes from the Atlantic Ocean. High background levels were found in the piranha Serrasalmus rhombeus (0.35 microg Hg x g(-1) muscle tissue, wet mass basis) and the peacock cichlid Cichla ocellaris (0.39 microg x g(-1)) from the Central Suriname Nature Reserve. Average mercury levels in freshwater fishes were higher in piscivorous species than in nonpiscivorous species, both in potentially contaminated water bodies (0.71 and 0.19 microg x g(-1), respectively) and in the control site (0.25 and 0.04 microg x g(-1), respectively). Mercury concentrations in piscivorous freshwater fishes were significantly higher in rivers potentially affected by gold mining than in the control site. In 57% of 269 piscivorous freshwater fishes from potentially contaminated sites, mercury levels exceeded the maximum permissible concentration of 0.5 microg Hg x g(-1). The highest mercury concentrations (3.13 and 4.26 microg x g(-1)) were found in two piranhas S. rhombeus from the hydroelectric reservoir Lake Brokopondo. The high mercury levels in fishes from Lake Brokopondo were to some extent related to gold mining because fishes collected at eastern sites (i.e., close to the gold fields) showed significantly higher mercury concentrations than fishes from western localities. In the estuaries, mercury levels in ariid catfish (0.22 microg x g(-1)) and croakers (0.04-0.33 microg x g(-1)) were distinctly lower than those in piscivorous fishes from contaminated freshwater sites. In the isolated Bigi Pan Lagoon, the piscivores snook Centropomus undecimalis (0.04 microg x g(-1)) and tarpon Megalops atlanticus (0.03 microg x g(-1)) showed low mercury levels. Mercury levels were significantly higher in marine fishes than in estuarine fishes, even with the Bigi Pan fishes excluded. High mercury concentrations were found in the shark Mustelus higmani (0.71 microg x g(-1)), the crevalle jack Caranx hippos (1.17 microg x g(-1)), and the barracuda Sphyraena guachancho (0.39 microg x g(-1)), but also in nonpiscivorous species such as Calamus bajonado (0.54 microg x g(-1)), Haemulon plumieri (0.47 microg x g(-1)), and Isopisthus parvipinnis (0.48 microg x g(-1)). Mercury levels were positively correlated with the length of the fish in populations of the freshwater piscivores S. rhombeus, Hoplias malabaricus, and Plagioscion squamosissimus, in estuarine species (Arius couma, Cynoscion virescens, and Macrodon ancylodon), and in S. guachancho from the Atlantic Ocean. PMID- 11437466 TI - Evidence of oxidative stress in American kestrels exposed to electromagnetic fields. AB - Exposure to electromagnetic fields (EMFs) alters melatonin, behavior, growth, and reproduction of captive American kestrels (Falco sparverius), particularly of males. EMF exposure is a "possible" human carcinogen and associated with some neurodegenerative diseases. Oxidative stress contributes to cancer, neurodegenerative diseases, and immune disorders. We tested whether EMF exposure elicits an avian immune response and alters oxidative stress levels. Captive male kestrels were bred under control or EMF conditions equivalent to those experienced by wild kestrels. Short-term EMF exposure (one breeding season) suppressed plasma total proteins, hematocrits, and carotenoids in the first half of the breeding season. It also suppressed erythrocyte cells and lymphocyte proportions, but elevated granulosa proportions at the end of the breeding season. Long-term EMF exposure (two breeding seasons) suppressed hematocrits in the first half of the reproductive period too. Results indicate that only short term EMF birds experience an immune response, particularly during the early half of the breeding season. The elevation of granulocytes, and the suppression of carotenoids, total proteins, and previously melatonin in the same kestrels, signifies that the short-term EMF male kestrels had higher levels of oxidative stress, due to an immune response and/or EMF exposure. Long-term EMF exposure may be linked to higher levels of oxidative stress through EMF exposure only. PMID- 11437468 TI - The complex of recombinant human insulin-like growth factor-I (rhIGF-I) and its binding protein-5 (IGFBP-5) induces local bone formation in murine calvariae and in rat cortical bone after local or systemic administration. AB - The influence of recombinant human insulin-like growth factor-I (rhIGF-I), its binding protein-5 (IGFBP-5) or their equimolar complexes on calvarial osteogenesis was investigated by quantitative radiography and histomorphometry after local administration to adult mice or mature rats. The systemic effects of these proteins were investigated in aged Sprague-Dawley rats with regard to their ability to prevent or restore bone mass in ovariectomy induced osteopenia as assessed by radiography, dual-energy X-ray absorptiometry (DEXA) analyses, peripheral computerized tomography (pQCT) and mineral analyses after daily s.c. administration for 3 or 8 weeks following a bone depletion period of 8 weeks. Bone mass of murine calvariae was significantly increased in a dose-dependent manner by the complex 7 days after discontinuation of local administration for 19 days in mice, whereas IGF-I alone expressed only weak effects. IGFBP-5 alone was ineffective in this respect. In the same model, only the complex had a weak osteogenetic potential in 7 week or 5 month old rats. Systemic long-term treatment with the complex of rhIGF-I/IGFBP-5 (2.0/7.6 mg/kg/day, s.c.) for 8 weeks resulted in significantly increased cortical thickness, area and mineral density in femoral midshaft or tibial metaphysis suggesting periosteal bone formation. This was obviously related to increased muscle strength since these effects were parallelled by increased body weight. No effect on trabecular bone occurred as demonstrated by site-specific analyses (vertebrae, proximal tibia) using DEXA, pQCT and radiography. This selective action of rhIGF-I/IGFBP-5 on periosteal bone formation is unique for an IGFBP. Femoral ash and calcium content, both corrected for tissue volume, increased slightly. However, when the increase in cortical thickness and bone mass was corrected for bone size, the effects are nearly abolished, suggesting an additional effect of bone growth. This potential deserves further evaluation in order to differentiate between effects on cortical bone via muscle strength and lack of efficacy on trabecular bone balance. PMID- 11437469 TI - Monoclonal antibodies to the carboxy-terminal Ea sequence of pro-insulin-like growth factor-IA (proIGF-IA) recognize proIGF-IA secreted by IM9 B-lymphocytes. AB - Insulin-like growth factor-I (IGF-I) circulates in human serum as a 7 kDa peptide but analysis of IGF-I cDNAs predicts two pro-hormone precursors (proIGF-IA and proIGF-IB) with distinct C-terminal E domains. The function of these precursors, and the E peptides generated on cleavage to mature IGF-I, is unknown, largely because of a lack of tools for distinguishing precursors from constituent peptides. We used a synthetic Ea peptide to develop monoclonal antibodies (MAbs) which can recognize the carboxy-terminal sequence of proIGF-IA. These were characterized using proIGF-IA generated by transfected HEK293 cells. The anti proIGF-IA MAbs immunoprecipitated two peptides (19--21 and 14 kDa) which were also recognized by MAbs to mature IGF-I. The proIGF-IA MAbs could also detect peptides of 9 and 4 kDa predicted to be Ea peptides. Treatment with N glycosidase proved the 19--21 kDa and 9 kDa bands to be glycosylated proIGF-IA and Ea peptide respectively. Using these antibodies, we have identified proIGF-IA secreted from the IM9 B-lymphocyte cell line. This work paves the way for studies on proIGF-IA and Ea peptide regulation and function. PMID- 11437470 TI - Multicentre collaborative study to calibrate IGF-II by bioassay and immunoassay: establishment of the First WHO Reference Reagent. AB - A preparation of recombinant insulin-like growth factor-II (IGF-II) (NIBSC code 96/538) was compared with local standards in bioassays and immunoassays by eight laboratories in four countries to assess its suitability for use as a World Health Organisation (WHO) reference reagent. Estimates of relative potencies for the bioassays gave a geometric mean of 1.04 (0.94--1.16) microg of local standard per microg of 96/538. Estimates of relative immunological activities by immunoassay gave a geometric mean of 1.15 (0.94--1.38) microg of local standard per microg of 96/538. The study provided evidence that a common standard for rhIGF-II would be helpful and that 96/538 was sufficiently stable to serve as a reference reagent. Accordingly 96/538 was established as the First WHO Reference Reagent for IGF-II, human, recombinant, and assigned a unitage of 5000 units per ampoule and on the basis of the immunoassay results a nominal mass content of 5 microg per ampoule. PMID- 11437471 TI - Developmental changes in the mRNA levels of IGF-I and its related genes in the reproductive organs of Japanese quail (Coturnix coturnix japonica). AB - Summary Insulin-like growth factor-I (IGF-I) is involved in the regulation of growth and differentiation of a variety of vertebrate tissues. The biological actions of IGF-I are mediated mainly by the IGF-I receptor (IGF-IR) and partly by the insulin receptor (IR) and modulated by IGF binding proteins (IGFBP). We conducted studies designed to clarify the possible roles of IGF system in the development of the avian reproductive organs. We cloned cDNAs of IGF-I, IGF-IR, IR and IGFBP-2 of Japanese quail and simultaneously measured the expression of these genes in the quail liver, testis and oviduct at different ages using a lysate RNase protection assay. Hepatic IGF-I mRNA levels increased rapidly and remained elevated during the rapid-growing period, which coincided with the period of rapid increase in testicular weight. IGF-I mRNA was detected at each stage of developing testis examined. Its level was high at the early stage and decreased with age. IGFBP-2 mRNA in testis exhibited a similar expression pattern to that of IGF-I, whereas a divergence in IGF-I and IGF-IR gene expression was observed. Both IGF-IR and IR mRNAs increased when the testis grew rapidly and decreased when sexual maturation was almost completed. These results suggest that IGF-I may serve as an autocrine/paracrine regulator as well as an endocrine regulator in the testicular development and function of Japanese quail. In the oviduct, IGF-I, IGF-IR, IR and IGFBP-2 mRNAs were also developmentally regulated. A rapid growth of the oviduct was accompanied by a significant increase in the level of IGF-I mRNA. The expression of genes encoding IGF-IR, IR and IGFBP-2 in the oviduct exhibited a similar developmental change to that of IGF-I. These results suggest that IGF-I mainly works in an autocrine and/or paracrine manner in the oviduct during the development of this organ. The findings of the present study provide further evidence of an important role for IGF system in the development and function of the avian reproductive system. PMID- 11437472 TI - Growth hormone (GH) and estradiol regulation of membrane-associated GH binding protein and GH receptors in GH releasing hormone transgenic mice. AB - Growth hormone (GH) releasing hormone (GHRH) transgenic mice were used to examine the influence of GH on GH receptor (GHR) and membrane-associated GH binding protein (MA-GHBP) levels by means of specific radioimmunoassays and Western blot analysis, since MA-GHBP was described as the major constituent of somatogenic binding to liver membranes in mice. In transgenic animals, a 10-fold increment over normal values was found for hepatic somatogenic binding that could be accounted for by a 3--4-fold increase in GHR and a 9-fold augmentation of MA-GHBP levels. The apparent molecular weight of MA-GHBP was smaller than that of serum GHBP, a difference that was partially abolished by endoglycosidase F digestion. In vivo treatment of female mice with 17 beta-estradiol led to an unexpected down regulation of MA-GHBP and GHR by 60--75% only in transgenic animals. MA-GHBP and GHR levels are strongly up-regulated by GH, although MA-GHBP up-regulation is much more important than that of GHR. PMID- 11437473 TI - The pharmacokinetics, pharmacodynamics, safety and tolerability following 7 days daily oral treatment with NN703 in healthy male subjects. AB - The aim of the present study was to assess the safety, pharmacokinetics and pharmacodynamics (including specificity) of NN703 (tabimorelin), a growth hormone (GH) secretagogue, in healthy male subjects following treatment for 7 days once daily. This was a randomized, double-blind and placebo-controlled study with four active dose levels: 1.71, 3.0, 4.5 and 6.86 mg/kg body weight. There was a dose related increase for GH area under the curve (AUC) (0-12 h) and GH C(max)(0--12 h); these were significantly higher on both days 1 and 7 as compared with placebo treatment (P = 0.04 to P< 0.0001); however, an overall significant decrease in GH release was found from day 1 to day 7 (P< 0.001). Insulin-like growth factor-I (IGF-I) and IGF binding protein 3 (IGFBP-3) increased at all dose levels (including placebo); however, a significantly higher increase as compared with placebo treatment was observed at the three highest dose levels for IGF-I (P = 0.04--0.0006) and at the highest dose level for IGFBP-3 (P = 0.03). There was no statistically significant increase in AUC (0-5 h) for follicle-stimulating hormone, luteinizing hormone and cortisol between active and placebo treatment for day 1 or 7. On day 1 only, a statistically significant increase in AUC (0--5 h) was found for prolactin at 1.71 and 6.86 mg/kg (P< 0.05), for thyroid stimulating hormone (TSH) at 3.0 mg/kg (P< 0.01) and for adrenocorticotrophic hormone (ACTH) at 4.5 mg/kg (P< 0.05); however, no dose--response relationship was observed for TSH or ACTH. In addition, a statistically significant decrease in AUC (0--5 h) for ACTH (3.0 and 6.86 mg/kg) and cortisol (1.71 mg/kg) was observed on day 7 (P< 0.05). Thus, NN703 is a promising candidate for treatment of absolute or relative GH deficiency. PMID- 11437474 TI - A new radioimmunoassay to measure rat insulin-like growth factor binding protein 4 (IGFBP-4) in serum, wound fluid and conditioned media. AB - Insulin-like growth factor binding protein-4 (IGFBP-4) is, like the other five IGFBPs, a critical regulator of the activity of insulin-like growth factor-I (IGF I) and IGF-II. Because of the possible role of IGFBP-4 in multiple systems including reproduction, pregnancy, bone formation and wound healing, we developed a radioimmunoassay (RIA) to measure the concentration of IGFBP-4 in the serum, extracellular fluid and conditioned media of cells of rodents so that these animal models could be better exploited. Rat IGFBP-4 was expressed in Escherichia coli and purified to homogeneity. The recombinant material was used to raise a rabbit polyclonal antibody against rat IGFBP-4 and for iodination and standards. The RIA developed was sensitive to less than 1 ng/mL of rat IGFBP-4 and IGF-I did not interfere. There was no cross-reactivity with other rat IGFBPs on immuno Western analysis of serum and wound fluid. IGFBP-4 from mouse serum did cross react in our assay; however, serum from horse, pig or human showed no cross reaction. Human IGFBP-1 and IGFBP-3 showed a very weak cross-reactivity. Serum IGFBP-4 levels showed no gender difference but did reveal a significant 66% increase in older rats. During the course of wound healing, which is IGF-I dependent, IGFBP-4 showed no changes. In conclusion an RIA for rat IGFBP-4 has been developed that specifically measures the concentration of IGFBP-4 in the sera, extracellular fluid and conditioned media of rodents. With this assay, the role of IGFBP-4 and IGFs in growth, development and the function of multiple systems can be further investigated using rodent models. PMID- 11437476 TI - Five images of maturity in adolescence: what does "grown up" mean? AB - This study focussed on the subjective meanings of maturity in adolescence, or what it means to adolescents to be grown up. Younger (6th grade) and older (9th grade) adolescents' descriptions (n=236) of their "grown-up" peers were examined through content analysis. This qualitative analysis revealed five images of maturity portrayed by adolescents: balanced maturity (adolescents who show psychosocial and behavioural maturity, and ability to balance work and play); an image focussed on privileges (adolescents who engage in problem behaviour and present what may be a facade of adult-like behaviour); an image focussed on responsibility (adolescents who may be psychosocially mature, but may have taken on inappropriately high levels of responsibility); an image focussed on power and status (adolescents who seem to have usurped an older status, by being bossy and controlling); and an image focussed on physical development (adolescents who show advanced levels of physical maturity). There were some gender and age differences in the frequencies of these five images. Discussion is directed at understanding the hallmarks of each image relative to scholarly notions of adult maturity. PMID- 11437477 TI - Adolescents' bereavement experiences. Prevalence, association with depressive symptoms, and use of services. AB - The present study set out to estimate the prevalence of bereavement experiences in adolescents, the association between these experiences and depressive symptoms and the attitudes of bereaved young people to professional interventions. The study was based on 1746 adolescents aged between 11 and 16 years from two secondary schools in Northern England. Questionnaire measures of bereavement experiences and depressive symptoms were completed by the adolescents in the classroom. One thousand three hundred and fifty-five (77.6%) reported that at least one of their first-or second-degree relatives or close friends had died. These losses were associated with increased levels of depressive symptoms in comparison with the loss of other relatives or pets. The impact of the loss of someone close depended to an important extent on the young person's perception of how the loss had changed their lives. Most adolescents did not feel the need for professional services. Those who did use these services had higher levels of depressive symptoms, suggesting that service use was likely to have been appropriate. PMID- 11437478 TI - Comparing adolescents' considerations for self-referral and counsellors' perceptions of these considerations: an exploratory study. AB - This study investigated the considerations of adolescents for self-referral to counsellors as reported by 421 senior-high school students and 123 school counsellors. Participants were asked to assess the extent to which different considerations affect adolescent willingness to self-refer to a counsellor. While the most important consideration for both samples was the counsellor's trustworthiness, some differences were found. Counsellors attributed more importance than adolescents to counsellor and adolescent gender and age, to adolescent academic and social status and to the conditions of the presenting problem, whereas the adolescents emphasized counsellor expertness. The results are discussed in terms of the strategies that can be implemented for motivating adolescents to independently approach counsellors with their problems. PMID- 11437479 TI - Young adolescents' wellbeing and health-risk behaviours: gender and socio economic differences. AB - In this paper we use the 1994-1997 Youth Surveys of the British Household Panel Study to examine the wellbeing of young adolescents. We conceptualize wellbeing as a multi-dimensional construct and we develop and test models of gender and age differences. Using confirmatory factor analysis, we find clear gender differences in self-esteem, self-efficacy, unhappiness and worries. We confirm that wellbeing and some health-risk behaviours (fighting and smoking) are linked. We test models that examine how family structure, father's occupation, tenure, and household income, affect adolescent wellbeing. While socio-economic factors affect health risk behaviours and also adolescents' reported worries, they have little impact on other aspects of youth wellbeing. The implications of these findings are discussed. PMID- 11437480 TI - Social anxiety and depression in adolescents in relation to perceived competence and situational appraisal. AB - The Social Anxiety and Phobia Inventory for Children (SPAI-C), the Children's Depression Inventory (CDI), Harter's (1982) Perceived Competence Scale for Children (PCSC), as well as an inventory of cost and likelihood appraisal of negative social and non-social events, were filled in by 184 adolescents (14-15 years old). It was expected that social anxiety would be specifically related to low perceived competence in the social domain and threat appraisal in the same domain, whereas depression would be related to more general perceived competence deficits. The relationships of social anxiety and depression to perceived competence and appraisal were largely supported. Social anxiety was most strongly related to perceived social competence as well as to both social cost and likelihood appraisals, whereas depression was more generally related to perceived competence, and appraisal of negative events did not improve the prediction of depression any more than the prediction based on perceived competence. PMID- 11437481 TI - The interaction of perceived maternal and paternal parenting styles and their relation with the psychological distress and offending characteristics of incarcerated young offenders. AB - A shortened form of the Parental Bonding Instrument (PBI) (Pederson, 1994) was used to examine the relationship between parenting styles and the psychological distress and offending patterns of a group of young male offenders held in custody in Scotland. High levels of psychological distress were linked with low parental care, but there was no association between psychological distress and parental control. Parental care was not a distinguishing factor in offending patterns, although high paternal control was linked with a younger age of first arrest. When interactions of paternal and maternal parenting styles were examined, young offenders who perceived poor parenting (i.e. neglectful parenting or affectionless control) from both parents had the highest levels of psychological distress overall. PMID- 11437482 TI - Our family, whose perspective? An investigation of children's family life and health. AB - Most quantitative studies of family life and family members' well-being rely on single informants in relation to both. This raises two issues; first, the validity of an individual's perceptions of family life, second, that part of any obtained association may arise because the same person provided information on both. This study addresses these methodological points, plus the substantive issue of whether family structure is related to child health/adjustment independently of family process. It employs reports of family life from 11-year olds and their parents, and of child health/adjustment obtained via self-report as well as from parents and teachers. Variance in child health/adjustment explained by family structure alone is extremely small. The strongest relationships between family process and child health/adjustment occur in respect of reports from the same informant, although some cross-informant relationships exist. Caution when interpreting studies of family life and well-being which rely on single informants is suggested, and the need for increased understanding of the reasons behind any obtained associations highlighted. PMID- 11437483 TI - The relationship of parenting style to self-reported mental health among two subcultures of Chinese. AB - The results of this investigation with 127 youths (aged 16-22 years) from mainland China supported the association of recalled parent's styles (assessed by the EMBU) with adolescents' self-evaluated health status (measured by the SCL 90). Many psychosomatic symptoms and lower scores on indexes of general mental health were significantly related to higher levels of parental rejection and denial, punishing tendency, overprotection and overinvolvement and to lower levels of parental emotional warmth and comprehension. The association between parenting style and mental health status of adolescents can be generalized across genders and across the two Chinese subcultures studied (Han and Kejia ethnical groups) with moderate to fairly high confidence. The mental health status of participants suggests some differences between different subcultures or genders on several variables of the SCL-90. PMID- 11437485 TI - Cytotoxic effect of an anti-liver monoclonal autoantibody obtained after neonatal thymectomy in mice. AB - A monoclonal autoantibody, LSA-1, against murine liver antigen was obtained by fusing spleen cells from a neonatally thymectomized BALB/c mouse with SP2/0 murine myeloma cells. The LSA-1 isotype was IgG2b and kappa. LSA-1 was specific to the liver, especially, to a liver-specific membrane lipoprotein (LSP) fraction. By Western blotting analysis, LSA-1 mainly detected a 100 kDa protein of LSP fraction. LSA-1 stained cytoplasm of the cryostat sections of liver in immunohistochemical analysis. Furthermore, the antigen recognized with LSA-1 was highly expressed on the surface of a murine hepatoma cell line, MH134, slightly on a murine normal liver cell line, C1469, and on freshly prepared hepatocytes, but not on spleen cells. LSA-1 had a cytotoxic activity on liver cell lines in the presence of a complement in vitro. Furthermore, injection of LSA-1 into mice induced liver injury. These results suggest that anti-liver autoantibody plays an important role in the induction of autoimmune hepatitis. Accordingly, this antibody will be a useful tool for the analysis of the pathogenesis of autoimmune hepatitis. PMID- 11437486 TI - DNA and heparin alter the internalization process of anti-DNA monoclonal antibodies according to patterns typical of both the charged molecule and the antibody. AB - The internalization into CHO-K1 fibroblasts of three polyreactive monoclonal IgG2a anti-DNA autoantibodies (mAbs), F14.6, J20.8 and F4.1, isolated from the same unimmunized (NZBxNZW) F1 mouse, and synthetic peptides derived from F4.1 was studied using a technique which quantifies nuclear accumulation. The localization of the mAbs was intranuclear. We compared the influence of two negatively-charged molecules, DNA or heparin. At low concentrations, DNA had dual effects-inhibitory or stimulatory-depending on the mAb. Heparin was inhibitory or had no effect. The possibility that proteoglycans are 'receptors' recognized by anti-DNA mAbs which bind through heparin-sensitive reactions, was explored. Only F4.1 internalization was partly inhibited in glycosaminoglycan-deficient cells. We propose that the complex alterations of internalization patterns of these polyreactive mAbs by the two negatively charged molecules can be explained by (a) the potential of polyreactive mAbs to bind to various charge (or conformation-) dependent 'receptors', (b) the potential of a subclass of mAbs complexed with DNA to utilize additional 'receptor(s)'. Glycosaminoglycans were required for internalization of F4.1-derived peptides, which remained extranuclear, suggesting that nuclear internalization of mAb F4.1 is a multistep process that requires certain sequences present on the intact mAb. PMID- 11437487 TI - B cells are selectively associated with thymic cortical but not medullary pathology in NZB mice. AB - Abnormal expansion of autoantibody-synthesizing B cells and self-reactive T cells, which most likely escape negative selection within the thymus, have both been characterized and reasoned to play a role in the pathogenesis of autoimmunity in NZB mice. Support for this thesis has been our observation that NZB mice have severe cortical and medullary thymic microarchitectural defects. As a means to dissect the roles of T and B cells in the induction of such abnormalities, B cell-deficient NZB mice were bred by backcrossing the Igh6(null)allele on to the NZB background (NZB-muMT mice). Such mice showed undetectable levels of autoantibodies. NZB-muMT mice, as compared to wild-type NZB mice, had lower absolute numbers of CD4(+)T cells. Furthermore, thymic abnormalities in NZB-muMT mice were restricted to the medulla. These data suggest that, while B cells may play a role in thymic cortical abnormalities, the medullary abnormalities are induced by other mechanisms. PMID- 11437488 TI - Increased expression of mXBP-1 (TREB-5) in thymic B cells in New Zealand mice. AB - New Zealand Black mice as well as several other murine models of murine lupus are well known for premature degeneration of thymus and development of autoimmunity. To focus on molecular events unique to murine lupus, we performed differential display using arbitrary primer pairs to distinguish NZB versus BALB/c thymus at 5 weeks of age. Following an extensive analysis of DNA bands that were either consistently up or downregulated and from studies of expression pattern of thymic genes by in situ nucleic acid hybridization, we focused on one clone that was consistently differentially expressed between NZB and BALB/c thymus. This clone was isolated, sequenced, and identified as the murine homologue of the human X box binding protein (hXBP-1), also known as TREB 5. mXBP-1 was found to be consistently upregulated in B cells in the thymic cortex of NZB and (NZBxNZW)F1, but not BALB/c, C3H/HeJ or C57BL/6 mice. In addition, it was dramatically elevated in MRL/ lpr but not MRL/++ mice; similarly, it was increased in BXSB/ Yaa male but not BXSB female thymic cortex. Of particular interest was an absence of mXBP-1 expression in the thymus of NZB/ Bln- Igh6(null)homozygotes. mXBP-1 has several putative functions, including the regulation of MHC class II expression and by virtue of its ability to recognize CRE-like elements shown to be involved in HTLV-1 transcription. PMID- 11437489 TI - A spontaneous canine model of mucous membrane (cicatricial) pemphigoid, an autoimmune blistering disease affecting mucosae and mucocutaneous junctions. AB - Mucous membrane pemphigoid (MMP) is a rare autoimmune blistering dermatosis of humans that was previously known as cicatricial pemphigoid. It is characterized by vesicles, ulcers and scarring that affect predominantly mucosae and mucocutaneous junctions. Circulating autoantibodies recognize epitopes on basement membrane proteins such as collagen XVII or laminin-5/6. Herein, we describe the clinico-pathological and immunological characteristics of 17 dogs afflicted with a dermatosis homologous to MMP of humans. Patients exhibited vesicles and erosions predominantly on mucous membranes or mucocutaneous junctions of the mouth, nose, eyes, genitalia or anus. Histopathology revealed subepithelial vesicles with variable dermal inflammation. Direct immunofluorescence demonstrated IgG or complement at the dermoepithelial junction. Indirect immunofluorescence using salt-split epithelia permitted the detection of circulating basement membrane-specific IgG autoantibodies in 15 cases. In 11 patients, autoantibodies recognized the NC16A segment of collagen XVII, as determined by salt-split indirect immunofluorescence, immunoblotting using canine keratinocytes and ELISA with synthetic canine peptides. In one dog, autoantiodies bound to the dermal side of salt-split epithelia and recognized epitopes within the 30 kDa carboxy-terminal segment of human collagen XVII. Canine MMP, like its human counterpart, exhibits distinctive clinical signs and histopathological lesions, yet circulating autoantibodies target different antigenic epitopes. This spontaneous canine model of MMP could prove useful for studies on the pathogenesis or therapy of this human disease. PMID- 11437490 TI - Evaluation of the percentage of peripheral T cells with two different T cell receptor alpha-chains and of their potential role in autoimmunity. AB - Approximately 25% of mature T cells possess two distinct cytoplasmic T cell receptor (TCR) alpha-chains, due to productive gene rearrangements of both alleles. Expression of two different alpha-chains at the cell surface is a potential risk factor for development of autoimmunity. However, it has been difficult to determine the frequency of peripheral T cells with two different alpha-chains at the surface. Our new approach is based on comparing by flow cytometry the percentage of cells that express a given Valpha-chain between wild type mice and mice that are hemizygous for a disrupted Tcra locus (Tcra+/-) and consequently unable to express two rearranged Tcra genes. We consistently found that approximately 8% of total peripheral T cells express two surface alpha chains. The importance of dual alpha-T cells in autoimmunity was examined in a mouse model for rheumatoid arthritis, namely collagen-induced arthritis (CIA). No significant difference was observed between Tcra+/- mice and wild-type littermates, considering arthritis incidence, day of disease onset, and maximum arthritic score. We therefore conclude that there is incomplete phenotypic allelic exclusion in TCRalpha, and that the presence of a significant number of potentially multireactive T cells does not increase the susceptibility to develop autoimmune arthritis. PMID- 11437491 TI - In vivo blockade of the Fas-Fas ligand pathway inhibits cyclophosphamide-induced diabetes in NOD mice. AB - There is compelling evidence to show that insulin dependent diabetes ensues from selective apoptosis of pancreatic beta-cells mediated by autoreactive T lymphocytes. The respective implication in this phenomenon of the various apoptotic pathways driven by Fas, perforin, or tumor necrosis factor is still ill defined. Here we took advantage of the cyclophosphamide-induced model of accelerated diabetes in NOD mice to explore the physiopathological role of the Fas-Fas Ligand pathway. A single injection of cyclophosphamide (200 mg/kg) to 7-8 week-old prediabetic NOD mice triggered diabetes within 10-15 days in 85-100% of the animals. Cyclophosphamide also induced a significant decrease in spleen T cells, that was most evident by days 6-10 after treatment, and selectively affected the CD3(+)CD62L(+)compartment that includes immunoregulatory T cells. To block the in vivo Fas-Fas ligand (Fas L) interaction we administered a biologically active recombinant fusion protein coupling mouse Fas to the Fc portion of human IgG1 (FAS-Fc). Mice treated with FAS-Fc (10 doses iv of 15 microg) starting on the day of cyclophosphamide injection up to day 22, were fully protected from disease. Unexpectedly this protective effect was not due to blockade of Fas-FasL-mediated beta-cell apoptosis but rather to the inhibition of the cyclophosphamide effect on T cells. Indeed FAS-Fc treatment prevented the drug-induced T cell depletion in general and that of immunoregulatory T cells in particular. Additionally, FAS-Fc administration limited to the phase of beta-cell destruction did not afford any protection. PMID- 11437492 TI - A transcriptional inhibitor of TNF-alpha prevents diabetes induced by multiple low-dose streptozotocin injections in mice. AB - The aim of the present study was to examine the effect of a transcriptional inhibitor of tumour necrosis factor-alpha (TNF-alpha) on the development of diabetes induced by multiple low doses of streptozotocin (STZ) in mice. MDL 201.449A is a novel transcriptional inhibitor of TNF-alpha gene expression and protein production and might therefore be potentially interesting in counteracting diabetes. We have studied the effect of MDL 201,449A on the development of hyperglycaemia and pancreatic insulitis in mice treated with multiple low-dose injections of streptozotocin. It was found that one daily sc injection of MDL 201,449A (25 mg/kg body weight) for 14 days prevented the development of hyperglycaemia following the streptozotocin injections. The mice treated with multiple low-dose injections of streptozotocin became gradually hyperglycaemic, and concomitant treatment with MDL 201,449A significantly reduced this elevation in blood glucose levels. In vitro MDL 201,449A reduced TNF-alpha mRNA levels dose-dependently by 75-80% after ConA stimulation of spleen cells, indicating the efficacy of MDL 201,449A to counteract TNF-alpha mRNA synthesis. These data suggest that transcriptional inhibition of TNF-alpha might be an interesting approach in the prevention of type 1 diabetes. PMID- 11437493 TI - An inhibitor of inducible nitric oxide synthase and scavenger of peroxynitrite prevents diabetes development in NOD mice. AB - Peroxynitrite (ONOO(-)) is a highly reactive oxidant produced by the interaction of the free radicals superoxide (O*-2) and nitric oxide (NO(*)). In a previous study, we found that peroxynitrite is formed in islet beta-cells of nonobese diabetic (NOD) mice. Here, we report that guanidinoethyldisulphide (GED), a selective inhibitor of inducible nitric oxide synthase (iNOS) and scavenger of peroxynitrite prevents diabetes in NOD mice. GED treatment of female NOD mice, starting at age 5 weeks, delayed diabetes onset (from age 12 to 22 weeks) and significantly decreased diabetes incidence at 30 weeks (from 80% to 17%). GED did not prevent pancreatic islet infiltration by leukocytes; however, beta-cells that stained positive for nitrotyrosine (a marker of peroxynitrite) were significantly decreased in islets of GED-treated mice (1+/-1%) compared with vehicle-treated mice (30+/-9%). In addition, GED significantly inhibited nitric oxide and nitrotyrosine formation and decreased destruction of beta-cells in NOD mouse islets incubated in vitro with the combination of proinflammatory cytokines interleukin 1-beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). These findings indicate that both superoxide and nitric oxide radicals contribute to islet beta-cell destruction in autoimmune diabetes via peroxynitrite formation in the beta-cells. PMID- 11437494 TI - Lack of detection of retroviral particles (HIAP-1) in the H9 T cell line co cultured with thyrocytes of Graves' disease. AB - Evidence for a possible aetiopathogenetic role of endogenous and/or exogenous retroviruses (RVs) in organ- and non-organ-specific autoimmune diseases is circumstantial in both humans and animal models. Intracisternal A type particles, antigenically related to HIV, have been reported in H9 cells co-cultured with homogenates of salivary glands obtained from patients with Sjogren syndrome and with synovial fluid of patients with rheumatoid arthritis. In order to identify a possible transfer of a putative 'infective RV agent' involved in the pathogenesis of human thyroid autoimmune disease, the H9 T cell line was co-cultured not only with thyroid homogenates, but also with viable thyrocytes, both prepared from glands of patients with Graves' disease. At the end of a prolonged co-culture period (24 weeks), no RV particles could be detected by electron microscopy in the H9 cells co-cultured with both thyroid preparations. These data seem to exclude the involvement of HIAP-1 in the aetiopathogenesis of human autoimmune thyroid disease. PMID- 11437495 TI - Multiple sclerosis: elevated expression of matrix metalloproteinases in blood monocytes. AB - Multiple sclerosis (MS) is an inflammatory, demyelinating disease of the central nervous system (CNS) characterized by blood-brain barrier (BBB) breakdown. Disruptions of BBB continuity result in an influx of activated T cells and monocytes, and could contribute to lesion formation in the CNS. Matrix metalloproteinases (MMP) are enzymes implicated in BBB disruption, and in degradation of extracellular matrix proteins and myelin components. An imbalance in levels of MMP and tissue inhibitors of MMP (TIMP) has been implicated in the pathogenesis of MS. Since monocytes form a major cell population in acute MS lesions and may facilitate their entrance into the CNS by secretion of MMP, knowledge on MMP expression by blood monocytes could be useful to improve our understanding of the pathogenesis of MS. In the present study, we examined the expression of MMP-1, -3, -7, -9, -14 and TIMP-1 mRNA by blood monocytes in patients with MS using in situ hybridization. Levels of MMP-1, -3, -7, -9 and of TIMP-1 mRNA expressing monocytes were elevated in MS compared to controls, while those of MMP-14 did not differ. We therefore conclude that MS is associated with elevated levels of MMP and TIMP expressing blood monocytes that may contribute to MS pathogenesis. PMID- 11437496 TI - Increased level of soluble HLA class I antigens in systemic lupus erythematosus: correlation with anti-DNA antibodies and leukopenia. AB - The concentration of soluble HLA class I (sHLA-I) was measured by ELISA in serum samples from 30 well-characterised SLE patients at high and low disease activity states and from 100 healthy controls. HLA-A allotypes in the patients were analysed by a PCR-based typing technique. A higher level of sHLA-I was found in SLE patient sera both at high and low disease activity than in controls (P< 0.001). The sHLA-I level was further increased during active disease (P< 0.01). Concentrations of sHLA-I correlated with anti-dsDNA antibodies at high disease activity, but not with disease activity as analysed by a modified SLEDAI. Numbers of leukocytes and lymphocytes, as well as levels of C1q and C3 correlated inversely with sHLA-I concentration. In five serial samples from ten patients the sHLA-I level co-varied with disease activity. Presence of HLA allotype A9 was associated with higher sHLA-I levels in both patients (P< 0.001) and controls (P< 0.001). We conclude that the increased sHLA-I concentration in SLE patients was related to several laboratory parameters reflecting disease activity suggesting that sHLA-I molecules are connected with the disease process. Increased sHLA-I level due to HLA-A allotype was not a disease susceptibility factor for SLE. PMID- 11437497 TI - Crossreactivity of human anti-dsDNA antibodies to phosphorylcholine: clues to their origin. AB - The presence of anti-double stranded DNA (dsDNA) antibodies is a serological diagnostic feature of systemic lupus erythematosus (SLE), an autoimmune rheumatic disorder. Studies by several investigators have suggested that a response to a microbial antigen can lead to the induction of SLE-like autoimmunity, in both humans and mice, since anti-dsDNA antibodies have been shown to crossreact with foreign antigens. In particular, anti-DNA antibodies have been shown to crossreact with phosphorylcholine (PC), a dominant epitope on pneumococcal cell wall polysaccharide. We have investigated the binding characteristics of human polyclonal anti-DNA antibodies from the sera of SLE patients. In this study we show that the DNA binding of polyclonal serum derived antibodies can be partially inhibited by phosphorylcholine (PC). The binding of affinity-purified anti-DNA antibodies from the sera of patients with SLE was also found to be inhibited by PC. We further demonstrated that the serum IgG1 (T dependent) anti-DNA response was more likely to crossreact with PC than the IgG2 (T independent) response to DNA. The studies suggest there may be a T dependent and T independent response to DNA with the T dependent response displaying more crossreactivity with microbial antigen. PMID- 11437498 TI - Distribution of porcine reproductive and respiratory syndrome virus in stillborn and liveborn piglets from experimentally infected sows. AB - Four pregnant sows were infected 3 weeks before their expected farrowing date with a Korean isolate (North American genotype) of porcine reproductive and respiratory syndrome virus (PRRSV). The distribution of virus in their stillborn and liveborn (killed 7 days after birth) offspring was assessed immunohistochemically and by in-situ hybridization. PRRSV antigen and nucleic acid were detected in lung, thymus, liver, tonsil, spleen, heart, kidney and lymph nodes from both stillborn and liveborn piglets. Positive cells typically exhibited a red (immunohistochemistry) or dark brown (in-situ hybridization) reaction product in the cytoplasm, without background staining. The most consistent labelling for PRRSV was in the thymus, tonsil and lymph nodes. The experiment suggested that, in prenatal piglets, PRRSV replicates primarily in lymphoid tissues, having gained access to them from the placenta via the bloodstream. PMID- 11437499 TI - Detection of European porcine reproductive and respiratory syndrome virus in porcine alveolar macrophages by two-colour immunofluorescence and in-situ hybridization-immunohistochemistry double labelling. AB - Two groups of five pigs aged 6 weeks were each infected oronasally with one of two different European isolates of porcine reproductive and respiratory syndrome virus (PRRSV). The animals were killed sequentially at 4, 7, 14 or 21 days post inoculation for examination. The methods used consisted of histopathology, and mono- and double-labelling techniques based on in-situ hybridization, immunofluorescence and immunohistochemistry. Porcine alveolar macrophages (PAMs) contained large amounts of PRRSV antigen and PRRSV RNA, as shown by double labelling with (1) either PRRSV immunofluorescence or PRRSV-specific in-situ hybridization with digoxigenin-labelled riboprobes, and (2) immunolabelling with Mac 387 antibody for calprotectin. Expression of PRRSV-RNA was not detectable in cytokeratin-positive hypertrophic and proliferating pneumocytes or in cells of alveolar ducts or bronchiolar epithelium. The use of two-colour immunofluorescence with confocal laser scanning microscopy and double labelling with in-situ hybridization-immunohistochemistry showed that PAMs were the only pulmonary target cells. This contradicts earlier reports that epithelial pulmonary cells may also be infected by PRRSV. PMID- 11437500 TI - Classical swine fever: pathogenesis of glomerular damage and immunocharacterization of immunocomplex deposits. AB - Twenty-six pigs were inoculated with a virulent isolate (Quillota strain) of classical swine fever (hog cholera) virus to determine the chronological development of lesions in the renal glomeruli and the pathogenesis of glomerular damage and immunocomplex deposition. The study included the use of histopathological, ultrastructural and immunohistochemical (detection of viral antigen gp55, myeloid-histiocyte antigen, IgM, IgG and C1q) techniques. The main changes in glomerular structure were observed from 7 days post-inoculation (dpi) onwards, at which time the glomeruli showed macrophage infiltrations in the mesangium, and viral infection in circulating cells, glomerular endothelial cells and podocytes. Moreover, significant subcellular changes were detected in podocytes, which appeared swollen, with fusion of foot processes. Immunocomplex deposits immunoreactive for IgM, IgG and C1q were detected in mesangial, subepithelial and subendothelial areas from 10 dpi, but viral antigen was not detected as a component of these deposits; fusion of foot processes had increased in severity, especially near immunocomplex deposits. All these changes had increased still further in the final phase of the experiment (14 dpi), with neutrophil infiltrations in the mesangium. PMID- 11437501 TI - A gross and microscopical morphometric evaluation of feline large intestinal anatomy. AB - The objective of this study was to examine measurable parameters that could be used to describe the gross and microscopical anatomy of the feline colon, which has not been previously characterized. Post-mortem data were collected from 35 specific pathogen-free cats. Gross morphometric data (total colonic length, wet weight, number of lymphoid aggregates) were collected together with microscopical measurement of crypt depth and numbers, intra-epithelial lymphocyte numbers and the number of proliferating cell nuclear antigen (PCNA)-positive cells and their relative position within the crypt. Colonic length as a percentage of intestinal length was remarkably constant (20.9+/-2.0%). Crypt depth, however, showed considerable inter-cat variation (149-688 microm); the crypts became deeper with distance from the anus. Cellular proliferation was predominantly in the lower part of the crypt, and the numbers of PCNA-positive cells increased with distance from the anus. The number of lymphoid aggregates varied with distance from the anorectum and appeared to reflect the bacterial load. Intra-epithelial lymphocytes were relatively sparse (3.9+/-2.7/100 epithelial cells); they showed considerable inter-cat variation but did not vary with distance from the anus. These data suggest that to improve the interpretation of morphometric parameters by reducing inter-cat variation, samples should be taken at a consistent distance from the anus, broad reference ranges having now been established. PMID- 11437502 TI - Invasion and spread of equine herpesvirus 9 in the olfactory pathway of pigs after intranasal inoculation. AB - The neuropathogenesis of equine herpesvirus 9 (EHV-9) in pigs was investigated by intranasal inoculation of the virus together with intramuscular administration of dexamethasone (DM). All infected pigs developed characteristic meningo encephalitis, accompanied by basophilic intranuclear inclusion bodies in the neuronal cells. One non-DM-treated and two DM-treated pigs had prominent malacic lesions in the rhinencephalon. Associated with the encephalitic lesions, there was invariably an increase in the number of nucleated cells in the cerebrospinal fluid (CSF). EHV-9 antigen was first detected in the nasal and olfactory epithelial cells in the nasal cavity, and in the neuroglial cells in the olfactory bulb. Subsequently it was demonstrated in the amygdaloid and caudate nuclei, and putamen. The virus was not isolated from the CSF. These results suggest that, after intranasal inoculation, EHV-9 replicates in the olfactory epithelial cells, spreading to the central nervous system via the olfactory pathway. PMID- 11437503 TI - A guinea-pig model of Hendra virus encephalitis. AB - Subcutaneous inoculation, but not intradermal (footpad) or intranasal inoculation, with high doses of Hendra virus (HeV) consistently produced disease in guinea-pigs. Of 15 subcutaneously inoculated animals, 14 developed vascular disease with positive HeV immunohistochemical labelling in a range of tissues. A new observation was the presence of lesions, including syncytial cells, with immunolabelling in the transitional epithelium of the bladder. Virus isolation from the urine rather than from nasal, oral, rectal or conjunctival swabs, the other external sites, was consistent with previous epidemiological work in horses, indicating a limited possibility of transmission. The dose used (30 000 to 50 000 TCID(50)), which was higher than in previous studies, produced microscopical lesions of encephalitis in eight of the 15 subcutaneously inoculated guinea-pigs, with positive immunolabelling in blood vessels and neurons, especially in the medulla, cerebellum and thalamus. The virus was recovered from six of the encephalitic brains. Severe vascular degeneration in the centres of encephalitic lesions in six of the eight encephalitic guinea-pigs and positive immunolabelling in the choroid plexus of a further animal indicated that the virus entered the brain following virus-induced vascular injury and choroid plexus invasion. Guinea-pigs would appear to be suitable for the study of HeV encephalitis. PMID- 11437504 TI - Oral inoculation of sheep with the agent of bovine spongiform encephalopathy (BSE). 1. Onset and distribution of disease-specific PrP accumulation in brain and viscera. AB - Sixty-three Romney sheep aged 6 months, consisting of three groups (PrP(ARQ/ARQ), PrP(ARQ/ARR), and PrP(ARR/ARR)genotypes) of 21 animals, were infected orally with brain tissue from BSE-infected cattle. Sub-groups of the 21 PrP(ARQ/ARQ) animals were killed, together with uninfected controls 4, 10, 16, 22 or 24-28 (after the development of full clinical disease) months post-inoculation (mpi). One sheep from each of the two groups of four killed at 4 or 10 mpi were shown by immunohistochemical examination to possess disease-specific PrP accumulations in single lymph nodes. At 16 mpi, such accumulations were detected in two of four infected sheep in some viscera and in the spinal cord and brain. At 22 mpi, three of five infected sheep had widespread disease-specific PrP accumulations in all tissues examined, but the remaining two animals gave positive results only in the central nervous system. Clinical disease appeared at 20-28 mpi. Three sheep killed with advanced clinical signs showed widespread PrP accumulation in brain, spinal cord and peripheral tissues. These results confirmed that PrP(ARQ/ARQ) Romney sheep are susceptible to experimental infection with the BSE agent. The different sites at which initial PrP accumulations were detected suggested that the point of entry of infection varied. Once established, however, infection appeared to spread rapidly throughout the lymphoreticular system. The results suggested that in some BSE-infected sheep neuroinvasion occurred in the absence of detectable PrP accumulations in the viscera or peripheral nervous system. In contrast to cattle with BSE, however, most sheep showed disease-specific PrP accumulations in the lymphoreticular system. In this respect, BSE-infected resembled scrapie-infected sheep; it is possible, however, that future research will reveal differences in respect of targeting of cell types within the lymphoreticular and peripheral nervous systems. The PrP(ARQ/ARR)and PrP(ARR/ARR)sheep were also killed in sub-groups at intervals after inoculation. Up to 24 mpi, however, none of these animals showed disease-specific PrP accumulations. Further results will be reported later. PMID- 11437505 TI - Natural paratuberculosis infection in rabbits in Scotland. AB - Natural paratuberculosis infection of rabbits (Oryctolagus cuniculus) was recently diagnosed in Scotland, and an investigation into the pathology of the disease in wild rabbits is reported in this paper. Evidence of Mycobacterium avium subsp. paratuberculosis (M.a. paratuberculosis) infection was detected in 22% of 110 rabbits; the organism was cultured from 17 of 110 rabbits, and histopathological lesions consistent with M.a. paratuberculosis infection were noted in 18 of 98 rabbits examined. No macroscopical lesions suggestive of M.a. paratuberculosis infection were observed. The histopathological lesions were either severe or mild. Severe lesions consisted of extensive macrophage granulomata and numerous giant cells, with many intracellular acid-fast bacteria in the small intestine. For the examination of formalin-fixed, paraffin wax embedded tissues, neither immunohistochemistry nor the polymerase chain reaction was as sensitive a method of diagnosis as histopathology. PMID- 11437506 TI - Comparative study of in-situ cell death induced by the viruses of viral haemorrhagic septicaemia (VHS) and infectious pancreatic necrosis (IPN) in rainbow trout. AB - The causative viruses of two diseases of rainbow trout, viral haemorrhagic septicaemia (VHS) and infectious pancreatic necrosis (IPN), exert much of their cytopathogenic effect in cell culture through the induction of apoptosis. In the present study, the TUNEL procedure was used to investigate the presence of apoptotic cells in different organs of rainbow trout infected with the viruses of VHS and IPN. VHS viral infection resulted in massive apoptosis in renal lymphoid tissue, where viral antigens were also detected. Large numbers of viral particles were observed in close proximity to apoptotic cells. Apoptosis was not detected in excretory cells of the renal tubules or in infected muscle cells. IPN virus did not induce apoptosis in the pancreas. However, the DNA degradation associated with apoptotic nuclei was observed in muscle lesions. Taken together, these results indicated that induction of apoptosis in vivo was critically influenced by the species of virus and the cell type. Moreover, it would seem likely that apoptosis contributed to the nature of the two diseases and to mortality. PMID- 11437507 TI - gammadelta T-cell lymphoma with tropism for various types of epithelium in a cow. AB - A 2-year-old Holstein cow developed multiple cutaneous masses, up to 10 cm in diameter, over the neck and trunk. The animal also had neoplastic lesions in internal organs, and epitheliotropism was observed not only in the skin but also in the conjunctiva, mammary glands, trachea, abomasum, small intestine, gall bladder, uterus and urinary bladder. Moreover, the neoplastic cells showed preferential homing to T-zones of lymphatic tissues. Because the lymphoma cells were positive for CD3 but not CD79a, and a few were CD2- or WC1-positive, this lymphoma was thought to be of gammadelta T-cell origin. Helicobacter -like organisms, found in the abomasum and small intestine, were considered to be associated with the severe neoplastic involvement of these organs, on the basis of the intense immunolabelling for major histocompatibility complex class II in the epithelial cells, and the presence of increased numbers of apparently normal intraepithelial T lymphocytes. PMID- 11437508 TI - Eosinophilia in duck embryos induced by an apathogenic strain of duck enteritis virus. AB - An apathogenic strain of duck enteritis virus was injected into the allantoic cavity of duck embryos at 17 or 18 days of incubation. Four to 6 days later, the embryos showed massive infiltration of eosinophilic granulocytes in the spleen. Peroxidase and immunohistochemical labelling showed that the granulocytes were peroxidase-positive and contained major basic protein, and were most likely eosinophils. UltraIstructural examination of the eosinophils demonstrated granules of various sizes and shapes, and granules within the granules. Electron microscopy also showed that splenic macrophages were capable of phagocytosis, indicating that duck embryos, even at 23-24 days of incubation, were capable of responding to infection. PMID- 11437509 TI - Lymphocytic neuritis of the ileum in sheep with naturally acquired and experimental paratuberculosis. AB - In a comparison of natural and experimentally induced ovine paratuberculosis, aggregations of mononuclear inflammatory cells were detected around nerves in the ileal submucosa in eight of 12 sheep with natural disease and in five of 14 animals with unequivocal experimental paratuberculosis. Such lesions were not seen in 10 other sheep that, despite attempted experimental infection, remained free from the disease, as judged clinically, histopathologically and by PCR assay. The lesions described resembled those observed in human leprosy. PMID- 11437510 TI - Classical swine fever: morphological and morphometrical study of pulmonary intravascular macrophages. AB - To gain further insight into the pathogenesis of classical swine fever (CSF), the changes induced by hog cholera (HC) virus in pulmonary intravascular macrophages (PIMs) were examined. Twelve pigs were inoculated by the intramuscular route with a virulent strain of HC virus (Quillota strain) and killed in groups of three at 4, 7, 10 and 14 days post-inoculation. Immunohistochemical and ultrastructural examination revealed HC virus infection in endothelial cells, PIMs, and interstitial and alveolar macrophages. In addition to viral replication, a predominant feature was the secretory activation of PIMs, characterized by expanded rough endoplasmic reticulum and hyperplastic Golgi complexes. The results obtained suggest that macrophage activation and the subsequent release of pro-inflammatory mediators play an important role in the pathogenesis of CSF. PMID- 11437511 TI - Susceptibility of rats of different ages to inoculation with swine haemagglutinating encephalomyelitis virus (a coronavirus) by various routes. AB - Haemagglutinating encephalomyelitis virus, strain 67N, was used to inoculate 1-, 2-, 4- and 8-week-old rats by the intracerebral (i.c.), intranasal (i.n.), intraperitoneal (i.p.), subcutaneous (s.c.), intravenous (i.v.) and oral routes with graded doses. The routes of infection, in descending order of efficacy, were: i.ci.ns.ci.pi.v. and oral. Rats aged 1 and 2 weeks were generally similar in terms of mortality and mean time to death, regardless of inoculation route, except for the oral route, which had little effect. In comparison with the 1- and 2-week-old rats, the 4-week-old rats were less susceptible to the virus by all routes. Eight-week-old rats inoculated by the i.ci.n. or s.c. routes died, but all those inoculated by other routes survived. To follow the spread of virus in the central nervous system, 4-week-old rats inoculated by the i.c. route were examined. The virus was first detected in the brain on day 1 and in the spinal cord on day 2. The viral titres in both tissues reached a plateau of 10(7) plaque forming units (PFU)/0.2 g by day 4, at which time clinical signs had developed. By immunohistochemical analysis, virus-specific antigen was found first in the pyramidal cells of the hippocampus and cerebral cortex, and later in the large sized neurons of the pons and spinal cord. Still later (day 4) immunolabelling was found in Purkinje cells of the cerebellum, but not in the ependymal cells, choroid plexus or other glial cells. PMID- 11437512 TI - Effects of lipopolysaccharide on a macrophage-like cell line (HS-P) from a rat histiocytic sarcoma. AB - Lipopolysaccharide (LPS) is a major modulator of macrophage functions. To characterize a newly established rat histiocytic sarcoma-derived cell line (HS P), immunophenotypic changes and cellular growth responses of HS-P cells exposed to LPS were investigated and compared with those of MT-9 cells isolated from a rat malignant fibrous histiocytoma. MT-9 cells have somewhat histiocytic features, because occasional cells react to rat macrophage-specific antibodies. Addition of LPS to cultured HS-P cells increased the numbers of cells immunopositive to ED1 (rat macrophage-specific antibody) and ED2 (rat histiocyte specific antibody) and stimulated the phagocytosis of latex beads, whereas LPS treated MT-9 cells did not show such immunophenotypic changes. LPS-treated HS-P cells showed enhanced immunolabelling of alpha-smooth muscle actin, suggesting a possible modulation of macrophages towards myofibroblastic cells. To evaluate cellular growth after the addition of LPS or fetal bovine serum, DNA synthesis was examined by measuring tritiated thymidine incorporation, and the mRNA expression of c- jun and c- myc (immediate early genes in the cell cycle) was examined by Northern blot analysis. In HS-P cells, the addition of serum greatly increased DNA synthesis and induced high expression of c- jun and c- myc; in contrast, LPS markedly depressed DNA synthesis and reduced the expression of c- jun and c- myc. HS-P cells were more sensitive than MT-9 cells to the growth promoting effect of serum and the growth-inhibiting effect of LPS. The study demonstrated that HS-P cells are highly LPS-responsive, indicating that they would be useful for studies of macrophage functions. PMID- 11437513 TI - Characterization of gross and histological lesions in Balb/c mice experimentally infected with herpesvirus saimiri 1 (HVS1). AB - Accidental B virus (Herpesvirus simiae) infection of human beings working with macaques is frequently fatal. However, the pathogenic potential of other similar simian alphaherpesviruses, such as the squirrel monkey virus Herpesvirus saimiri (HVS1), is virtually unknown. As part of an effort to develop a murine model for infections with these agents, Balb/c mice were inoculated intramuscularly in the left hindlimb with 10 to 10(6) plaque forming units (PFU) of HVS1. After observation for clinical signs of infection for 21 days, mice were killed and specimens collected for serology and histopathology. Mice receiving 510(3) PFU of HVS1 exhibited severe, pruritic, ulcerative skin lesions near the site of inoculation and developed unilateral or bilateral hindlimb paralysis with severe muscle atrophy. Histological lesions were characterized by a necrotizing dermatitis and folliculitis. Spinal cord lesions consisted of a non-suppurative myelitis affecting primarily the ipsilateral dorsal horn of the thoracolumbar spinal cord with occasional extension to ventral and contralateral spinal cord regions. Immunohistochemical labelling confirmed the presence of viral antigen within the lesions, and anti-HVS1 IgG concentrations were related to the occurrence of disease. HVS1 infection in some mice extended from the ipsilateral dorsal horn and funiculus into the ventral and contralateral grey and white matter, resulting in bilateral hindlimb paralysis. Thoracolumbar spinal cord lesions resolved without continued spread of the virus to cranial nervous system structures, i.e., cervical spinal cord and brain. PMID- 11437514 TI - Expression of the apxIV gene in pigs naturally infected with Actinobacillus pleuropneumoniae. AB - The apxIV gene was detected, by in-situ hybridization with a non-radioactive digoxigenin-labelled probe, in formalin-fixed, paraffin wax-embedded samples of lung tissue from 10 pigs naturally infected with Actinobacillus pleuropneumoniae. A 442 base pair DNA probe of the apxIV gene from A. pleuropneumoniae serotype 2 was generated by the polymerase chain reaction. All 10 pigs infected with A. pleuropneumoniae serotypes 2, 5, 6, or an untypable strain showed a distinct, positive signal in the degenerate alveolar leucocytes in alveolar spaces, and in the dense zone of degenerated cells in granulation tissue surrounding the necrotic areas. Thus, the study demonstrated the presence of the apxIV gene in pleuropneumonic lesions caused by A. pleuropneumoniae. PMID- 11437515 TI - Detection of immune system cells in paraffin wax-embedded ovine tissues. AB - This report describes a method (fixation, paraffin wax-embedding and immunolabelling) for the demonstration of several immune system cell epitopes (CD1, CD2, CD4, CD8, CD14, CD21, CD45R, WC-1, 28 kDa surface antigen, immunoglobulins and MHC II antigens) in ovine lymph nodes collected at necropsy. Cell surface epitopes considered to be sensitive to processing methods were successfully demonstrated by a procedure that included the use of a non-aldehyde containing, zinc salts-based fixative, coupled with a sensitive system of immunolabelling. This novel method had the advantage of avoiding antigen retrieval steps and of providing consistently good morphological definition. PMID- 11437516 TI - Onset and distribution of tissue prp accumulation in scrapie-affected suffolk sheep as demonstrated by sequential necropsies and tonsillar biopsies. AB - Tonsillar biopsies (single or multiple) or necropsies, or both, were performed on sheep taken from a Suffolk flock in which frequent cases of scrapie had occurred over a period of several years. Clinically affected sheep of the susceptible PrP(AQ/AQ)genotype had widespread disease-specific PrP accumulation in the central nervous system (CNS), lymphoreticular system and peripheral ganglia. In nine healthy PrP(AQ/AQ)Suffolk sheep between 4 and 7 years of age, PrP could not be demonstrated post mortem in any of the lymphoreticular tissues, or in the peripheral ganglia or CNS. Tonsillar biopsies taken from animals of the resistant PrP(AR/AR)and PrP(AR/AQ)genotypes at age 3, 8, 14, 20 or 26 months did not show PrP accumulation. Disease- specific PrP accumulation in tonsillar biopsies from PrP(AQ/AQ)sheep was not seen in 20 animals aged 3 months, but was found in two of 10 animals at age 8 months and in eight of 10 animals at age 20 months. The numbers of PrP-positive tonsillar biopsies obtained from sheep previously biopsied on more than one occasion was greater than the number of positive tonsils obtained from other susceptible sheep of comparable ages. The earliest disease-specific PrP accumulation seen was in tingible body macrophages within germinal centres and only later was it detected in cells resembling follicular dendritic cells. Fourteen PrP(AQ/AQ)sheep examined post mortem at up to 17 months of age and which had not previously been biopsied or were biopsied only once had no CNS or tonsillar PrP accumulations. Two of these sheep subjected to necropsy at 14 months had PrP accumulation in lymphoreticular tissue, where it was confined to the mesenteric lymph nodes. In susceptible sheep, only low levels of immunohistochemically detectable PrP were present in a minority of follicles from tonsillar biopsies of young lambs, but by 14 months of age widespread PrP accumulation, affecting many or even all follicles, was present. Although clinical cases had widespread PrP accumulations in viscera, susceptible survivors had no such accumulations in tissues of the lymphoreticular system, peripheral nervous system or CNS, suggesting that some animals were not exposed to infection or were exposed to a non-infectious dose. PMID- 11437517 TI - Neosporosis in Mexican dairy herds: lesions and immunohistochemical detection of Neospora caninum in fetuses. AB - Of 211 aborted bovine fetuses collected from Mexican dairy herds between January 1996 and March 1999, 73 showed microscopical lesions consistent with neosporosis. Of these 73 fetuses, 58 (79%) showed lymphocytic myocarditis, 39 (53%) showed microgliosis and multifocal necrosis in the brain, 39 (53%) showed lymphocytic hepatitis, and 19 (26%) showed lymphocytic myositis. Immunohistochemical examination of brain, myocardium and liver from 53 of the same 73 fetuses demonstrated Neospora caninum antigens in 41 (77%), of which 19 (46%) gave positive results in one of the three sites, 15 (37%) in two, and seven (17%) in three. The results indicated the presence of neosporosis in a number of the main dairy farming regions of Mexico. PMID- 11437518 TI - Evaluation of a rapid western immunoblotting procedure for the diagnosis of bovine spongiform encephalopathy (BSE) in the UK. AB - Bovine brain tissue samples from 625 UK cattle, clinically suspected as bovine spongiform encephalopathy (BSE) cases, were used in a blind analysis to assess a rapid Western immunoblotting technique (Prionics Check; Prionics AG, Zurich), which detects bovine disease-specific protease-resistant prion protein (PrP(Sc)). By means of statutory histopathological examination, 599 of the 625 cattle were confirmed as BSE cases by the demonstration of spongiform encephalopathy, the remaining 26 being classified as negative. Duplicate samples from the same animals were also examined by electron microscopy for the presence of abnormal brain fibrils (scrapie-associated fibrils; SAFs). The Prionics technique showed a high sensitivity, particularly when compared with the fibril detection test; the detection rates were 99.3% and 92.0% respectively, with histopathology being used as the "gold standard". The false negative results by the Prionics test were possibly related to the sampling procedure. Analysis of 50 BSE-positive samples revealed similar glycoprofiles, the majority of PrP(Sc)isoforms being di glycosylated protein. The Prionics test also detected PrP(Sc)in the four brain samples from the 26 histopathologically negative animals, apparently reducing the specificity of the test to 84.6%; however, confirmatory positive results in these samples were obtained by demonstrating SAF or by immunohistochemical examination, or both. It was concluded that the Prionics test detected PrP(Sc)in a small percentage (0.64%) of clinically suspected BSE cases showing no spongiform change. Since January 2000, the Prionics Western blot test has been introduced as one of the statutory tests for the diagnosis of clinically suspected BSE and scrapie cases in the UK. PMID- 11437519 TI - Experimental Elaphostrongylus alces infection in goats. AB - Five goats aged 4 months were each inoculated with approximately 300 third-stage larvae of Elaphostrongylus alces, and killed for post-mortem examination after 14 150 days. No clinical signs of disease were observed during the experiment. Pathological examination revealed that the larvae penetrated the walls of the abomasum and small intestine and migrated towards the caudal vertebral canal. However, the great majority of larvae were apparently destroyed along the migratory route, and development to adult parasites in the vertebral canal was not seen. During migration, the larvae caused focal inflammation and necrosis in the gastrointestinal wall, liver, mesentery and lungs. The study suggests that the only effect of E. alces infection on goats is the formation of focal visceral lesions during abdominal larval migration; it also confirms the infectivity of E. alces for domestic ruminants. PMID- 11437520 TI - Demonstration of chymotryptic and tryptic activities in mast cells of rodents: comparison of 17 species of the family Muridae. AB - On the basis of studies in laboratory rats, mast cells were originally classified into two subgroups, namely, mucosal mast cells (MMCs), which contained chymase, and connective tissue mast cells (CTMCs), which contained both tryptase and chymase. This classification has been applied to other animal species, despite the fact that the MMCs and CTMCs of such species sometimes consist of mixed populations of mast cells in terms of tryptase and chymase constitution. This report describes the protease constitution of mast cells in 17 species of nine genera (Acomys, Apodemus, Cricetulus, Meriones, Millardia, Mus, Rattus, Sigmodon and Vandeleuria) of the family Muridae. MMCs with negative tryptase activity were detected only in the intestinal mucosa of six subspecies of Mus musculus, two Rattus spp. and Vandeleuria oleacea, and only Apodemus sylvaticus possessed CTMCs with no tryptase activity. Since mast cells conforming to the conventional classification were observed only in three of the nine genera examined, we propose that mast cells of rodents of the family Muridae should be classified by their protease constitution rather than by their location. PMID- 11437521 TI - Bone and ivory points in the Lower and Middle Paleolithic of Europe. AB - The existence of shaped bone and ivory points, to be used as awls or with wooden hafts, has been suggested for the Lower Paleolithic sites of Torralba and Ambrona and for several Middle Paleolithic sites, such as Vaufrey, Combe Grenal, Pech de l'Aze I and Camiac. The use of hafted bone and ivory points would imply a spear armature technology similar to that well documented in the Upper Paleolithic, often considered an innovation introduced to Europe by anatomically modern humans. The controversial ivory points from the two Spanish sites, whose fracture morphology is considered natural by G. Haynes (1991), have been reanalyzed, checking for putative traces of human manufacture and utilization as described by Howell & Freeman (1983), i.e., polish, flaking of stem, ground edges, striations from manufacture and contact with a haft or binding. We have been able to study 19 new proboscidean tusk tips from the ongoing Ambrona excavations by a Spanish team. For these and nine other Middle Paleolithic bone and antler points we use optical and SEM microscope analysis, taphonomic analysis, comparative observations of Upper Paleolithic bone points, experimental observations of manufacturing traces, modern tusk samples, and data on several bone and antler pseudo-points from carnivore accumulations. We show that none of the objects we have studied can be interpreted as an intentionally shaped point. The absence of hafted bone points in the Middle Paleolithic of Europe is contrasted with evidence of the use of hafted stone points since OIS 5 or earlier in Eurasia and Africa. We suggest that the absence of organic spear armatures in the Middle Paleolithic is not due to a deficiency in the technology of Neandertals but may be tied to the organizational strategies of the hunters and to patterns of game choice and capture. PMID- 11437522 TI - The Plio-Pleistocene vegetation and climate of Sterkfontein and Swartkrans, South Africa, based on micromammals. AB - Micromammalian remains from Oldowan and Acheulean horizons in Sterkfontein Member 5E (M5E-O and M5E-A) and Swartkrans Members 1-3 (SKX1-3) provide information for the period between about 2 and 1 Ma. Another small sample from Sterkfontein Member 4 (M4) refers to a short period around 2.8-2.6 Ma and material from Sterkfontein post Member 6 infill (post-M6) relates to the Middle Stone Age occupation around 100 ka. Virtual absence of evidence of digestion on in situ lower molars of the major prey species, Mystromys albicaudatus, indicates that one of the owls Tyto alba, T. capensis or Asio capensis was the responsible predator. Over-emphasis on riverine grassland supports this suggestion, as does faunal resemblance between the Sterkfontein Valley samples and modern prey samples for the three owl species concerned. The location of the roost and the range of prey species point to the barn owl T. alba. There is no indication of differential bias between samples. South African species of eagle owl (Bubo spp.) are ruled out. All samples appear to represent interglacial conditions. The Grassland-Savanna ecotone probably lay nearby, as did the border between Moist and Arid Savanna. Possible modern analogues occur about 200 km west and southwest of the valley more than 250 m lower in altitude. South of Mafikeng all three vegetation types intergrade but the rainfall is higher than that postulated for some of the units in the Sterkfontein Valley. Similar climatic conditions occur near Kuruman and Kimberley where Arid Savanna merges into Grassland. Landscape information indicates a succession from riverine grassland, sometimes with Acacia trees, through hillsides with bush, grass and some trees, to plains with open savanna woodland. The suggested climate for the Sterkfontein Valley varied between +/-310 mm and possibly 550 mm mean annual precipitation, summer aridity index 3.8-4.1 and percentage winter rainfall 23-24. Minimum and maximum monthly temperatures would have been higher than they are today but with the temperature range reduced. PMID- 11437523 TI - Hominid skull fragments from Late Pleistocene layers in Leine Valley (Sarstedt, District of Hildesheim, Germany). AB - Three cranial fragments were recovered from coarse-grained deposits dug up by a suction dredge from gravel pits on the Leine river flats in the vicinity of Sarstedt (northwestern Germany). Also recovered were a number of artefacts which, upon careful inspection, could be assigned to the Middle Paleolithic. The geological pattern of the Leine Valley in this region suggests that these fragments were deposited in the lower terrace during a yet undetermined warm period-possibly Brorup or Odderade-during the Weichsel glaciation. However, attribution to the Eemian period or a Saale interstadial cannot be ruled out. The features of the Sarstedt (Sst) I infant temporal are known from Neanderthals (e.g., Weimar-Ehringsdorf, Engis, Krapina 1) and can be seen in specimens from the European late- Homo erectus group as well. Subadult individuals do not always exhibit full development of features characteristic for adults and-to some extent anticipate the succeeding developmental stage (i.e., neoteny). The Neanderthal autapomorphies characterizing the fragments of the occipital and the parietal are certainly consistent with assigning both unequivocally to the species H. neanderthalensis. The presence of Middle Paleolithic artefacts recovered from the same deposits are commensurate with the presence of Neanderthals. However, there is no clear contextual association of any archaeological and fossil human material. Future DNA research will hopefully add up to the established morphological picture. PMID- 11437524 TI - Locomotor behavior of Lagothrix lagothricha and Ateles belzebuth in Yasuni National Park, Ecuador: general patterns and nonsuspensory modes. AB - Field study of the locomotor behavior of sympatric woolly monkeys (Lagothrix lagothricha) and spider monkeys (Ateles belzebuth) in undisturbed rainforest of northern Ecuador revealed similar patterns in use of plant forms (categorized tree and liana structure), and substantial differences in the frequencies of use of different grouped modes (aggregates of kinematically similar specific modes). Lagothrix progressed more than Ateles by leaping/dropping and quadrupedal walking/running, whereas Ateles exhibited more suspensory locomotion. Grouped modes are associated with different plant forms in similar ways in the two species. In contrast, the species differed in use of tree zone (trunk/bole, major branches, intermediate branches, and terminal branches), with Lagothrix using intermediate branches and Ateles terminal branches more. Correlated with this difference was greater use by Lagothrix of quadrupedal movement, especially on intermediate branches, and greater use of suspensory modes by Ateles, especially in the terminal zone. Further research is needed to determine how these patterns are facilitated and constrained by morphological mechanisms. Analysis of specific locomotor modes within groups shows several interspecific differences in relative frequencies. PMID- 11437525 TI - Adult honeybee's resistance against Paenibacillus larvae larvae, the causative agent of the American foulbrood. AB - American foulbrood is a widespread disease of honeybee larvae caused by the spore forming bacterium Paenibacillus larvae subsp. larvae. Spores represent the infectious stage; when ingested by a larva they germinate in the midgut. The rod shaped vegetative forms penetrate the larva's intestinal tissue and start multiplying rapidly, which finally kills the larva. Spores fed to adult honeybees, however, do not harm the bees. We investigated this phenomenon. Specifically, we studied the influence of the adult honeybee midgut on the vegetative growth and on the germination of spores of P. larvae larvae. We focused on two groups of adult workers that are likely to have large numbers of spores in their gastrointestinal tracts in infected colonies: middle-aged bees, which are known to remove or cannibalize dead larvae and clean brood cells, and winterbees, which do not have frequent chances to defecate. We found that midgut extract from winterbees and worker-aged bees of different colonies almost completely inhibited the growth of the vegetative stage of P. larvae larvae and suppressed the germination of spores. The inhibiting substance or substances from the adult midgut are very temperature stable: they still show about 60% of their growth-inhibiting capacity against this bacterium after 15 min at 125 degrees C. We established a method to test growth-inhibiting factors against P. larvae larvae in vitro. PMID- 11437526 TI - Purification and characterization of a natural agglutinin in the hemolymph of the prawn Penaeus indicus H. Milne Edwards. AB - A natural agglutinin in the hemolymph of the marine prawn Penaeus indicus was isolated by gel filtration chromatography, purified using polyacrylamide gel electrophoresis, and characterized. Prawn agglutinin has a native molecular mass of 181 kDa and consists of two monomeric units (97 and 84 kDa), maintains some agglutinating activity over a wide pH range (7-9), and is inactivated at 85 degrees C. The agglutinin was denatured upon mixing with trichloroacetic acid, phenol, chloroform, and 45% ammonium sulfate. It was also sensitive to trypsin digestion. The results indicate that prawn agglutinin is proteinaceous in nature, with agglutinating, hemolytic, and antibacterial properties against marine bacteria and erythrocytes with carbohydrate binding sites. PMID- 11437527 TI - Developmental temperature effects on five geographic isolates of the entomopathogenic nematode Steinernema feltiae (Nematoda: Steinernematidae). AB - The development of five geographic isolates of Steinernema feltiae at 5, 8, 10, 15, 20, 25, and 28 degrees C in wax moth, Galleria mellonella, larvae was examined. The isolates were from Mediterranean (Sinop from Turkey, SN from France, and Monterey from California), subtropical (Rafaela from Argentina), and tropical (MG-14 from Hawaii) regions. All isolates caused 100% mortality of wax moth larvae and developed and produced progeny between 8 and 25 degrees C. At 28 degrees C, mortality was 100%, but no progeny was observed. The highest infective juvenile production was observed at 15 degrees C for all isolates. In general, the tropical isolate, MG-14, had the lowest production of infective juveniles. The time of emergence of the infective juveniles from the host cadaver showed some differences among isolates, with the Sinop isolate having the earliest emergence time from cadavers at 15 degrees C (10 days) and 20 degrees C (8 days). At 25 degrees C, the infective juveniles of the Sinop, SN, and Rafaela isolates emerged from the cadavers from 5 to 7 days. Time of host death by all isolates showed no differences at 8, 10, 15, 20, and 28 degrees C. At 25 degrees C for all isolates (except the MG-14), shorter times to host death were observed. Host death occurred at 12 days at 8 degrees C, 9 to 11 days at 10 degrees C, 4 to 5 days at 15 degrees C, 3 days at 20 degrees C, and 2 days at 25 and 28 degrees C. For penetration efficiency, the Sinop, SN, and Rafaela isolates penetrated their hosts at 5, 8, and 10 degrees C. Penetration of the infective juveniles was consistently high for all isolates at 15, 20, 25, and 28 degrees C, but it was significantly lower for the MG-14 isolate at 15, 25, and 28 degrees C. No progeny production occurred at 28 degrees C, but nematode penetration did occur with the MG-14 isolate having significantly lower penetration than the other isolates. When nematodes were produced at 8, 15, and 23 degrees C in wax moth larvae, all isolates had infective juveniles with longer body lengths at 8 degrees C followed by 15 and 23 degrees C. To further verify body length at the different temperatures, beet armyworm, Spodoptera exigua, larvae and dog-food agar medium were used, respectively, for in vivo and in vitro culture of the Sinop isolate. Infective juvenile body length showed the same trends, with the longest being at 8 degrees C and decreasing in length from 15 to 23 degrees C. The data suggest that quality of food for the nematode and temperature (that is, developmental time) influence the body length of the infective juvenile. PMID- 11437528 TI - Wolbachia bacteria effects after experimental interspecific transfers in terrestrial isopods. AB - Wolbachia bacteria are intracellular parasites, vertically transmitted from mothers to offspring through the cytoplasm of the eggs. They manipulate the reproduction of their hosts to increase in frequency in host populations. In terrestrial isopods for example, Wolbachia are responsible for the full feminization of putative males, therefore increasing the proportion of females, the sex by which they are transmitted. Vertical transmission, however, is not the only means for Wolbachia propagation. Infectious (i.e., horizontal) transmission between different host species or taxa is required to explain the fact that the phylogeny of Wolbachia does not parallel that of their hosts. The aim of this study was to investigate, by experimental transinfections, whether Wolbachia strains could be successfully transferred to a different, previously uninfected isopod host. While Wolbachia survived in all the studied recipient species, vertical transmission was efficient only in cases where donor and recipient species were closely related. Even in this case, Wolbachia strains did not always keep their ability to entirely feminize their host, a deficiency that can be link to a low bacterial density in the host tissues. In addition, Wolbachia infection was associated with a decrease in host fertility, except when the bacterial strain came from the same host population as the recipient animals. This suggest that Wolbachia could be adapted to local host populations. It therefore seems that isopod Wolbachia are highly adapted to their host and can hardly infect another species of hosts. The successful infection of a given Wolbachia strain into a new isopod host species therefore probably requires a strong selection on bacterial variants. PMID- 11437529 TI - Effects of time, temperature, and honey on Nosema apis (Microsporidia: Nosematidae), a parasite of the honeybee, Apis mellifera (Hymenoptera: Apidae). AB - Newly emerged adult bees were fed with Nosema apis spores subjected to various treatments, and their longevity, proportions of bees infected, and spores per bee recorded. Spores lost viability after 1, 3, or 6 months in active manuka or multifloral honey, after 3 days in multifloral honey, and after 21 days in water or sugar syrup at 33 degrees C. Air-dried spores lost viability after 3 or 5 days at 40 degrees, 45 degrees, or 49 degrees C. Increasing numbers of bees became infected with increasing doses of spores, regardless of their subsequent food (active manuka honey, thyme honey, or sugar syrup). Final spore loads were similar among bees receiving the same food, regardless of dose. Bees fed with either honey had lighter infections than those fed with syrup, but this may have been due to reductions in their longevity. Bees fed with manuka honey were significantly shorter lived, whether infected or not. PMID- 11437530 TI - Pathological and physiological changes in the South African freshwater crab Potamonautes warreni calman induced by microbial gill infestations. AB - The impact of microbial gill infestations on the pathology and physiology of the freshwater crab Potamonautes warreni was investigated by comparison of infested and uninfested crab populations from, respectively, a polluted and an unpolluted site along the Mooi River, North West Province, South Africa. Heavy gill infestations by bacteria (70%), peritrichous ciliates such as Lagenophrys sp. (15%), Zoothamnium sp. (10%), and Epistylis sp. (5%), and motile protozoans resulted in species-specific lesions in the gill epithelia of P. warreni and physiological changes in crabs from the polluted site. Bacterial colonies enmeshed in polysaccharide-like films produced indentations of the gill cuticular surfaces and dissociation of microvillous membranes at the basal zone of epithelial cells of gill lamellae of P. warreni. Lagenophrys sp. induced large subcuticular spaces with an unfolding or resorption of the plasma membrane in the gill epithelia. The attachment of stalks of Zoothamnium and Epistylis resulted in dilation of lamellar tissues, the formation of vacuoles, and an increase in subcuticular spaces in the epithelia. Physiological changes in infested crabs included significant differences (P = 0.001) in increments of wet body mass and a reduced growth rate over time compared with uninfested crabs. The specific oxygen consumption (M(O2)) in rested infested crabs significantly increased (31.29 +/- 5.8 micromol O2/kg/min) compared with the M(O2) in uninfested crabs (27.92 +/- 5.6 micromol O2/kg/min; P = 0.009). The heart rate of infested rested P. warreni was significantly lower (40.77 +/- 13.79 beats/min; P < 0.02) than that in uninfested crabs (61.09 +/- 29.02 beats/min) but the heart rate of infested crabs increased significantly with body mass (r = 0.53, P = 0.02). These findings suggest an interrelationship among organic pollution, microbial gill infestations, and specific pathological and physiological responses in the crab host. The role of P. warreni and its microbial gill communities as bioindicators of pollution are discussed. PMID- 11437531 TI - Effects of in vitro passage of Beauveria bassiana on virulence to Bemisia argentifolii. PMID- 11437532 TI - A possible Phasmarhabditis sp. (Nematoda: Rhabditidae) isolated from Lumbricus terrestris (Oligochaeta: Lumbricidae). PMID- 11437533 TI - Adult Diabrotica spp. (Coleoptera: Chrysomelidae) infection at emergence with Iindigenous Beauveria bassiana (Deuteromycotina: Hyphomycetes). PMID- 11437534 TI - Effect of host insects on activation of Entomophaga maimaiga resting spores. PMID- 11437535 TI - Acidic pH-induced contractile dysfunction via downstream mechanism: identification of pH-sensitive domain in troponin I. PMID- 11437536 TI - Localization, anchoring, and functions of protein kinase C isozymes in the heart. AB - Although protein kinase C (PKC) was identified more than 20 years ago, and is involved in a wide variety of essential cellular processes, assigning specific roles to each PKC isozyme has proved difficult. Results over the last few years have suggested that much of the specificity of activated PKC isozymes is attributed to their subcellular localization bringing them into close proximity to a subset of substrates. Our laboratory has taken advantage of the importance of PKC localization and studied the way in which PKC isozymes are anchored. We have identified PKC anchoring proteins (RACKs or Receptors for Activated C Kinase) and used information about interaction sites between PKC isozymes and their respective RACKs to design peptides which modulate translocation of specific PKC isozymes to the functional site. These isozyme-specific peptides can be delivered into isolated or cultured cells or expressed in transgenic mice to determine the role of specific PKC isozymes in particular functions. Here we will describe the isozymes-specific peptide activators and inhibitors that we have developed and the specific functions of each isozyme in cardiac ventricular tissue. PMID- 11437537 TI - Localization of regions of troponin I important in deactivation of cardiac myofilaments by acidic pH. AB - Ca2+-activation of cardiac muscle myofilaments is more sensitive to depression by acidic pH than is the case with skeletal myofilaments. We tested the hypothesis that this difference is related to specific regions of the TnI (troponin I) isoforms in these muscles. We exchanged native Tn complex in detergent-extracted fiber bundles from mouse ventricles with Tn containing various combinations of fast (fsTnI) or slow skeletal (ssTnI) complexed with either cardiac TnC (cTnC) or fsTnC, and with cTnC complexed with the following chimeras: (1) fsTnI N-terminal region (fN) plus cTnI inhibitory peptide (cIp) and cTnI C-terminal region (cC); and (2) cTnI N-terminal region (cN)-cIp-fsTnI C-terminal region (fC). We determined the change in half maximal Ca2+(DeltaEC50) for tension activation at pH 7.0 and pH 6.5. Similar DeltaEC50 values were obtained for unextracted controls (5.53+/-0.30 microm), for preparations containing cTnI-cTnC (5.74+/-0.40 microm), and preparations exchanged with cTnI-fsTnC (5.63+/-0.40 microm). However, replacement of cTnI with fsTnI significantly decreased DeltaEC50 to 3.95+/-0.17 microm. Replacement of cTnI with ssTnI also significantly depressed DeltaEC50 to 2.07+/-0.15 microm. Results of studies using the chimeras demonstrated that the C-terminal domains of cTnI and fsTnI are responsible for these differences. This conclusion also fits with data from experiments in which we measured Ca2+-binding to the regulatory site of cTnC in binary complexes containing cTnC with cTnI, fsTnI, or the chimeras. Our results localize a region of TnI important in effects of acidosis on cardiac myofilaments and extend our earlier data indicating that C-terminal regions of cTnI outside the Ip are critical for activation by Ca2+. PMID- 11437538 TI - Influence of embryonic cardiomyocyte transplantation on the progression of heart failure in a rat model of extensive myocardial infarction. AB - Cell transplantation has been proposed as a future therapy for various myocardial diseases. It is unknown, however, whether the encouraging results obtained in animal models of ischemia and reperfusion, cryoinjury or cardiomyopathy can be reproduced in the setting of permanent coronary artery occlusion and extensive myocardial infarction (MI). Embryonic cardiac cells were isolated and cultured for 3 days to confirm viability, morphology and to label cells with BrdU or the reporter gene LacZ. Seven days after extensive MI, rats were randomized to cell (1.5x10(6)) transplantation (n=11) or culture medium injection (n=16) into the myocardial scar. Echocardiography study was performed before and 53+/-3 days after implantation to assess left ventricular (LV) remodeling and function. During follow-up, there was no mortality among cell-treated rats v 4 of 16 control rats (P=0.12). X-gal staining, BrdU and alpha -SMA immunohistochemistry identified the engrafted cells 1 week, 4 weeks and 8 weeks after transplantation, respectively. Antibodies against alpha -SMA, connexin-43, fast and slow myosin heavy chain revealed grafts in various stages of differentiation in 10 of 11 cell treated hearts. Many of them, however, kept their embryonic phenotype and were isolated from the host myocardium by scar tissue. Serial echocardiography studies revealed that cell transplantation prevented scar thinning, LV dilatation and dysfunction while control animals developed scar thinning, significant LV dilatation accompanied by progressive deterioration in LV contractility. Transplantation of embryonic cardiomyocytes after extensive MI in a rat model attenuate LV dilatation, infarct thinning, and myocardial dysfunction. Still, many grafts remain isolated and do not differentiate into an adult phenotype, even when studied 2 months after grafting. PMID- 11437539 TI - Overexpression of tumor necrosis factor- alpha activates both anti- and pro apoptotic pathways in the myocardium. AB - We have previously reported that mice with cardiac-specific overexpression of tumor necrosis factor (TNF)- alpha develop myocardial inflammation, cardiac hypertrophy, and dilated cardiomyopathy. TNF- alpha is reported to induce apoptosis in cultured cardiac myocytes. To investigate the role of apoptosis in this transgenic model, wild-type controls (WT) and transgenic mice (TG) at the age of 1, 8, and 40 weeks were analyzed. Increased incidence of apoptosis in TG was indicated by DNA laddering. TUNEL assays revealed that the frequencies of apoptotic cells were increased in the TG myocardium at all ages. However, as revealed by histochemical and immunofluorescent methods, most of the apoptotic cells appeared to be non-myocytes even in the mice with overt congestive heart failure. To elucidate the signaling pathways responsible for TNF- alpha induced apoptosis, expression of apoptosis-related genes were evaluated by multi-probe RNase protection assays. Transcripts for death-domain-related proteins, including TNFR1, Fas, FADD, TRADD, and RIP, were constitutively expressed in WT and upregulated in the TG myocardium. Expression of caspase-1 through -8 was also enhanced in TG. While both anti- and pro-apoptotic Bcl-2 family genes were constitutively expressed in WT, TNF- alpha overexpression strongly induced anti apoptotic A1 in the myocardium. Furthermore, TNF- alpha overexpression activated NF- kappa B, a mediator of anti-apoptotic pathways, in the myocardium. Thus, overexpression of TNF- alpha activated both anti- and pro-apoptotic pathways in the myocardium, resulting in an increase of apoptosis, primarily in non-myocytes. These results suggest that TNF- alpha by itself is not sufficient to induce apoptosis in cardiac myocytes in vivo. PMID- 11437540 TI - Gender influences on sarcoplasmic reticulum Ca2+-handling in failing human myocardium. AB - Gender has recently been implicated as an important modulator of cardiovascular disease. However, it is not known how gender may specifically influence the Ca2+ handling deficits that characterize the depressed cardiac contractility of human heart failure. To elucidate the contributory role of gender to sarcoplasmic reticulum (SR) Ca2+ cycling alterations, the protein levels of SR Ca2+-ATPase (SERCA), phospholamban, and calsequestrin, as well as the site-specific phospholamban phosphorylation status, were quantified in a mixed gender population of failing (n=14) and donor (n=15) myocardia. The apparent affinity (EC50) and the maximal velocity (Vmax) of SR Ca2+-uptake were also determined to lend functional significance to any observed protein alterations. Phospholamban and calsequestrin levels were not altered; however, SERCA protein levels were significantly reduced in failing hearts. Additionally, phospholamban phosphorylation (serine-16 and threonine-17 sites) and myocardial cAMP content were both attenuated. The alterations in SR protein levels were also accompanied by a decreased V(max)and an increased EC50 (diminished apparent affinity) of SR Ca2+-uptake for Ca2+ in failing myocardia. Myocardial protein levels and Ca2+ uptake parameters were then analyzed with respect to gender, which revealed that the decreases in phosphorylated serine-16 were specific to male failing hearts, reflecting increases in the EC50 values of SR Ca2+-uptake for Ca2+, compared to donor males. These findings suggest that although decreased SERCA protein and phospholamban phosphorylation levels contribute to depressed SR Ca2+-uptake and left ventricular function in heart failure, the specific subcellular alterations which underlie these effects may not be uniform with respect to gender. PMID- 11437542 TI - Enhancement of the T-type calcium current by hyposmotic shock in isolated guinea pig ventricular myocytes. AB - It is known that swelling and shrinkage of cardiac cells can modulate their electrical activity. However, the effects of osmotic manipulation on cardiac T type calcium current (I(CaT)) has not been previously reported. In this study, we have examined the effects of cell swelling on I(CaT), using the whole cell patch clamp configuration. Isolated guinea-pig ventricular myocytes were swollen by an external hypotonic challenge (0.7 T). We found that I(CaT)is enhanced during a hypotonic shock. This current has been determined to be the T type calcium current since it is rapidly activated and inactivated, its threshold was at negative potentials and was blocked by 40 microm Ni2+. Disruption of microfilaments by cytochalasin D and of microtubules by colchicine prevented the activation of I(CaT)during cell swelling. Taxol had no effect. These results indicate that I(CaT)is increased during cell swelling and this effect needs an intact cytoskeleton. PMID- 11437541 TI - Bradykinin B2 receptor is involved in the late phase of preconditioning in rabbit heart. AB - Activation of bradykinin B2 receptor has been shown to confer short-term cardioprotection against a prolonged ischemic insult. The present study was designed to delineate the role of B2 receptor in the late phase of ischemic preconditioning. Anesthetized, open chest, male rabbits were assigned to 1 of 6 groups (n=8/group). Ischemic preconditioning was elicited by four 5-min occlusion periods interspersed with 10 min of reperfusion. To test the role of B2 receptors, rabbits were pretreated with specific receptor antagonist, HOE-140 (1 microgm/kg IV bolus), 15 min prior to ischemic preconditioning. Additionally, two separate groups of animals were treated by intra-atrial infusion with either bradykinin (0.05 microg/kg/min for 15 min) or saline. Twenty-four hours later, the animals were subjected to 30 min of ischemia and 3 h of reperfusion. Infarct size was determined by tetrazolium staining. Ischemic preconditioning reduced infarct size from 43.09+/-4.66 to 20.65+/-1.87 (% risk area, P<0.05), which was blocked by HOE-140 as indicated by increase in infarct size (36.72+/-4.04%, P<0.05). HOE-140 treatment had no significant effect on infarct size in the sham group. Similarly, intra-atrial infusion of bradykinin caused decrease in the infarct size from 52.36+/-2.17% in the saline control group to 22.83+/-1.71% (P<0.05). The degree of infarct limitation with bradykinin was comparable to ischemic preconditioning (20.65+/-1.87%v 22.83+/-1.71%, P>0.05). For the first time, these results provide evidence for the involvement of B2 receptor in the genesis of late phase of ischemic preconditioning. PMID- 11437543 TI - Modulation of the extracellular divalent cation-inhibited non-selective conductance in cardiac cells by metabolic inhibition and by oxidants. AB - The effect of metabolic inhibition and oxidative stress on the monovalent cation permeable, extracellular divalent cation-inhibited non-selective conductance was investigated in ventricular myocytes at 22 degrees C. Under whole-cell voltage clamp, with L-type Ca2+ channels blocked by nifedipine, and K+ currents blocked by Cs+ substitution for K+, removal of Ca2+(o)and Mg2+(o) induced a non-selective current (I(NS-(Ca)o)) in mouse, rabbit and rat cells. Removal of glucose increased I(NS-(Ca)o)in the absence of Ca2+(o) and Mg2+(o), but failed to induce this current in the presence of the divalent cations. Further inhibition of glycolysis by 2-deoxyglucose (DOG; 10 mM, in zero glucose) or of mitochondrial function by rotenone (10 microM) or NaCN (5 mM) also failed to induce I(NS (Ca)o)in the presence of Ca2+(o) and Mg2+(o). Even when given together, DOG and rotenone did not induce I(NS-(Ca)o) in the presence of divalent cations. Preactivated I(NS-(Ca)o) was increased by the oxidants thimerosal (50 microM), diamide (500 microM) and pCMPS (50 microM). However, none of these drugs nor NEM (1 mM) did elicit I(NS-(Ca)o)in the presence of Ca2+(o) and Mg2+(o). Exposure of rat myocytes to Ag+ induced a current resembling I(NS-(Ca)o) (reversing at -5 mV; blocked by 100 microM Gd3+) even in the presence of divalent cations. The data indicate that metabolic inhibition only regulates activated I(NS-(Ca)o)but does not induce the opening of closed channels, and that small oxidants like Ag+ may induce I(NS-(Ca)o) activation by accessing at sites unavailable for larger molecules. PMID- 11437546 TI - Analysis of the Interacting Octad System of (12)CH(4). AB - The octad system of methane (12)CH(4) has been analyzed with a fourth-order (459 parameters) Hamiltonian for energy levels and a third-order (28 parameters) dipole moment expansion for infrared intensities. Nearly 8000 assigned positions and 2500 measured line intensities (obtained with the Fourier transform spectrometer located at Kitt Peak National Observatory/National Solar Observatory) have been considered in the modeling. Infrared (Octad-Ground State) and hot bands (Octad-Dyad) data have been used. While the model does not reproduce the measurements at their inherent experimental precisions, it has been sufficient to interpret and substantially assign this spectral region of methane for the first time in over 30 years of investigation. Copyright 2001 Academic Press. PMID- 11437547 TI - The C(2)Pi(r)-X(2)Sigma(+) Transition of BO. AB - The C(2)Pi(r)-->X(2)Sigma(+) electronic transition of BO has been observed in emission under high resolution. Thirty-six bands have been analyzed for (11)BO and six for (10)BO. The C state is shown to be a regular state and better constants are given. Strong perturbations are discussed on the basis of the theoretical calculations of Karna and Grein (J. Mol. Spectrosc. 122, 356-364 (1987)). Copyright 2001 Academic Press. PMID- 11437548 TI - Identification of the High-Lying Pi Component of NiF by Laser-Induced Fluorescence. AB - The laser-induced fluorescence excitation spectrum of jet-cooled NiF has been studied in the range 435-570 nm. Eight electronic transitions, involving four high-lying Pi components and two lower states, the ground state X(2)Pi(3/2), and the low-lying state [0.25](2)Sigma, respectively, have been recorded. Two new Pi components are identified for the first time and their rotational constants are determined. The [20.4]Pi(3/2) component is assigned as the v'=1 level of the previously known [19.7](2)Pi(3/2) state. The [20.3]Pi(3/2) component may be linked to the component of an unreported quartet state. The previously reported [20.0](2)Pi(1/2) component is reassigned as a subcomponent of (2)Delta. Copyright 2001 Academic Press. PMID- 11437549 TI - Diode-Laser Spectroscopy: Line Profiles of H(2)O in the Region of 1.39 &mgr;m. AB - Using a stabilized diode-laser spectrometer, we have studied the line profiles of water vapor near 7185.59 cm(-1). With low pressures of pure gas (90% purity after a single isolation step at yields of 8 mg/L of insect cell medium (purified fusion protein). The recombinant GPIbalpha fragment was shown to be posttranslationally sulfated and glycosylated, although its glycosylation differed from that of the equivalent GPIbalpha fragment isolated from human platelets. The differential glycosylation, however, did not affect the function of the recombinant GPIbalpha fragment in either von Willebrand factor (vWf) or thrombin binding as these were both found to be identical to those of the same-length GPIbalpha fragment derived from human platelets. The calmodulin tag was also exploited in the development of assays to measure directly vWf and thrombin binding, since it did not interfere with either, demonstrating the feasibility for the use of this soluble receptor fusion protein in detailed biophysical assays to investigate the molecular mode of binding of platelet glycoprotein Ibalpha to these ligands. PMID- 11437596 TI - Characterization of Itk tyrosine kinase: contribution of noncatalytic domains to enzymatic activity. AB - Itk is a Tec family tyrosine kinase found in T cells that is activated upon ligation of the T cell receptor (TCR/CD3), CD2, or CD28. Itk contains five domains in addition to the catalytic domain: pleckstrin homology, Tec homology which contains a proline-rich region, Src homology 3, and Src homology 2. To provide a basis for understanding the contribution of these various domains to catalysis, recombinant Itk was purified and its substrate specificity determined by steady-state kinetic methods. Measurements of the rates of phosphorylation of various protein substrates, including Src associated in mitosis 68K protein (SAM68), CD28, linker for activation of T cells, and CD3 zeta, at a fixed concentration indicated that SAM68 was phosphorylated most rapidly. Wild-type Itk and three Itk mutants were characterized by comparing their activity (k(cat)) using the SAM68 substrate. A deletion mutant removing the pleckstrin homology domain and part of the Tec homology domain (Itk(Delta152)) had approximately 10 fold less activity than wild type, a mutant with an altered proline-rich domain (P158A,P159A) had a more dramatic 100-fold loss of activity, and the catalytic domain alone was essentially inactive. Itk(Delta152) had K(m) values for ATP and SAM68 nearly identical to those of the wild-type enzyme, while Itk(P158A,P159A) had approximately 3-fold higher K(m) values for each substrate. SAM68 phosphorylation by the wild-type and mutant enzymes in the presence of several tyrosine kinase inhibitors were compared using a homogeneous time-resolved fluorescence assay. Both the Itk(Delta152) deletion mutant and the Itk(P158A,P159A) mutant had IC(50) values similar to those of the wild-type enzyme for staurosporine, PP1, and damnacanthal. These comparisons, taken together with the similar K(m) values for ATP and SAM68 substrate between the wild-type and the mutant enzymes, indicate that the amino acids in the N-terminal 152 residues and proline-rich domains enhance catalysis by affecting turnover rate rather than substrate binding. PMID- 11437597 TI - Characterization of horse cytochrome c expressed in Escherichia coli. AB - We have expressed horse cytochrome c in Escherichia coli. The gene was designed with E. coli codon bias and assembled by using a recursive polymerase chain reaction method. The far-ultraviolet and near-ultraviolet/Soret circular dichroism (CD) spectra show that the structure of recombinant horse cytochrome c is the same as that of the authentic protein. CD-detected thermal denaturation studies were used to measure the thermodynamic parameters associated with two state denaturation. The free energy of denaturation for the recombinant protein is 10.0 +/- 2.3 kcal mol(-1) at pH 4.6 and 25 degrees C, which agrees with the value for the authentic protein. The expression system will help advance our understanding of the roles of cytochrome c in electron transfer, oxidative stress, and apoptosis by allowing the production of protein variants. PMID- 11437598 TI - Overexpression and refolding of the hydrophobic ribosomal P0 protein from Trypanosoma cruzi: a component of the P1/P2/P0 complex. AB - The P0 protein is part of the ribosomal eukaryotic stalk, which is an elongated lateral protuberance of the large ribosomal subunit involved in the translocation step of protein synthesis. P0 is the minimal portion of the stalk that is able to support accurate protein synthesis. The P0 C-terminal peptide is highly antigenic and a major target of the antibody response in patients with systemic lupus erythematosus and patients suffering chronic heart disease produced by the Trypanosoma cruzi parasite. The T. cruzi P0 (TcP0) protein was cloned into the pRSET A vector and expressed in Escherichia coli fused to a His-tag. The identity of the protein was confirmed by immunoblotting. Due to the formation of inclusion bodies the protein was purified using the following steps: (i) differential centrifugation to separate the inclusion bodies from soluble proteins and (ii) affinity chromatography under denaturing conditions. TcP0 showed high tendency to aggregation during refolding assays. However, TcP0 could be efficiently folded in the presence of a low concentration of SDS. The folding of the protein was confirmed using urea gradient electrophoresis, limited proteolysis, circular dichroism, and tryptophan fluorescence. Native electrophoresis showed that the folded TcP0 (and not a folding intermediate) was the cause of aggregation in the absence of SDS. The protocol described here permitted us to obtain large amounts (up to 30 mg per culture liter) of pure and folded TcP0, a very hydrophobic protein with a high tendency to aggregation. PMID- 11437599 TI - Mammalian glycosyltransferase expression allows sialoglycoprotein production by baculovirus-infected insect cells. AB - The baculovirus-insect cell expression system is widely used to produce recombinant mammalian glycoproteins, but the glycosylated end products are rarely authentic. This is because insect cells are typically unable to produce glycoprotein glycans containing terminal sialic acid residues. In this study, we examined the influence of two mammalian glycosyltransferases on N-glycoprotein sialylation by the baculovirus-insect cell system. This was accomplished by using a novel baculovirus vector designed to express a mammalian alpha2,6 sialyltransferase early in infection and a new insect cell line stably transformed to constitutively express a mammalian beta1,4-galactosyltransferase. Various biochemical assays showed that a foreign glycoprotein was sialylated by this virus-host combination, but not by a control virus-host combination, which lacked the mammalian glycosyltransferase genes. Thus, this study demonstrates that the baculovirus-insect cell expression system can be metabolically engineered for N-glycoprotein sialylation by the addition of two mammalian glycosyltransferase genes. PMID- 11437600 TI - Expression and one-step purification of intracellular human prolactin in insect cells. AB - Human prolactin was expressed in insect culture cells by recombinant baculoviruses carrying prolactin gene cDNA placed under the transcriptional control of polyhedrin gene promoter of Autographa californica nuclear polyhedrosis virus. Preliminary results of recombinant human prolactin expression as extracellular as well as intracellular product of baculovirus expression system were presented at the FEBS Meeting in Nice, France, in 1999 (Abstracts, p. 288). In the present work prolactin was expressed as a hexahistidine-tagged fusion protein and recombinant protein was purified by metal affinity resin. Yields varied between approximately 20 and 35 mg/liter of medium. This recombinant prolactin was biologically active in Nb2 lymphoma cell proliferation assay and after simple purification could substitute for pituitary-derived prolactin. PMID- 11437601 TI - Purification of a cysteine proteinase from Carica candamarcensis L. and cloning of a genomic putative fragment coding for this enzyme. AB - We describe the purification of a cysteine proteinase from latex of Carica candamarcensis, hereby designated CC23. The enzyme has been purified by ion exchange chromatography and behaves electrophoretically as a monomer of M(r) 23,000 and optimal pH of 8.0. It displays a basic isoelectric point, has one cysteine residue in the active site by titration with E-64, confirmed by DNA sequencing, and responds to proteinase inhibitors as a classic cysteine proteinase. The K(m) and k(cat)/K(m) for CC23 using BAPNA were respectively 14.7 +/- 1.8 x 10(-4) M and 1.3 x 10(3) M(-1) s(-1). Therefore, the catalytic efficiency of CC23 is sixfold higher than that of CC-I, another proteinase from the same plant. DNA primers were designed to amplify by PCR a genomic sequence related to this enzyme. An 895-bp DNA fragment was cloned and sequenced. It shows strong homology with chymopapain isoform IV from C. papaya. The translated sequence is similar to that of chymopapain isoform II (73%) and CC-III (77%) from C. candamarcensis. PMID- 11437602 TI - [W206R]-procaspase 3: an inactivatable substrate for caspase 8. AB - We report here the cloning and high-level expression of a soluble proform of human caspase 3 (Ser(24)-H(277)) engineered to contain a short stretch of N terminal sequence (MTISDSPREQD) from the prosegment of procaspase 8 and a C terminal heptahistidine tag. The precursor protein isolated from extracts of recombinant Escherichia coli by immobilized metal-ion affinity chromatography was predominantly unprocessed and migrated as a 32-kDa polypeptide on sodium dodecyl sulfate-polyacrylamide gels. Incubation of this protein with recombinant human caspase 8 produced fragments characteristic of the properly processed caspase 3, but the product was inactive. Amino-terminal sequence analysis of the caspase 3 polypeptides proved that caspase 8 had specifically cleaved the Asp(175)-Ser(176) bond to yield the expected p18 and p12 subunits, with partial cleavage at the Asp(28)-Ser(29) bond to release the prosegment. The lack of caspase 3 activity was found to be the result of a fortuitous mutation in which Trp(206) in the S4 subsite was replaced by arginine (W206R). This mutant procaspase 3, which we call m-pro3, serves as a useful reagent with which to test the efficacy of caspase 8 inhibitors in blocking processing of the natural polypeptide substrate of this enzyme and may be valuable as a source of "proenzyme" for crystallographic analysis. PMID- 11437603 TI - Single-step method for purification of Shiga toxin-1 B subunit using receptor mediated affinity chromatography by globotriaosylceramide-conjugated octyl sepharose CL-4B. AB - A new single-step purification method for Shiga toxin (Stx) was developed using receptor-mediated affinity chromatography, in which Gb3Cer (globotriaosylceramide) was conjugated to octyl Sepharose CL-4B as a carrier. This method achieves high yield and high purity in a small column on which Gb3Cer has been immobilized at high density. Using this affinity column, the Stx1 B subunit was purified with homogeneity by a one-step procedure from a crude extract of recombinant Stx1 B subunit-producing Escherichia coli. The purified Stx1 B subunit conserved a natural pentamer structure confirmed by gel filtration and sedimentation equilibrium analysis. Furthermore, the purified Stx1 B subunit was able to bind specifically to Gb3Cer expressed on Burkitt's lymphoma cells. This versatile purification method can be used to isolate various types of natural as well as recombinant Stx, facilitating fundamental studies of human diseases caused by this toxin. PMID- 11437604 TI - Purification of a bacterially expressed herpes simplex virus type 1 origin binding protein for use in posttranslational processing studies. AB - The origin binding protein (OBP) encoded by the UL9 open reading frame of herpes simplex virus type 1 (HSV-1) plays an essential role in productive infection by promoting the initiation of viral DNA synthesis. In this study, OBP was inducibly expressed in Escherichia coli and purified to homogeneity using a two-step chromatographic separation procedure. The properties of this recombinant OBP (rOBP) were found to be indistinguishable from those of the virus-encoded protein. Since rOBP was synthesized in bacterial cells, it lacked the posttranslational processing which normally occurs in OBP produced in HSV-1 infected mammalian cells and could therefore be exploited in experiments which addressed the effects of protein modification on OBP function. As an initial study, the impact of phosphorylation on enzymatic activity was examined using rOBP which had been treated with a panel of purified cellular kinases. rOBP was found to act as a substrate for nearly all of the kinases tested in (32)P-labeled phosphate transfer assays. However, only phosphorylation by protein kinase A (PKA, or cAMP-dependent protein kinase) was shown to significantly alter the enzymatic properties of rOBP, as it increased by five- to eightfold the ATPase activity associated with this protein. Activation of this critical viral DNA replication enzyme by a cAMP-dependent kinase such as PKA may be of some relevance in the natural course of HSV-1 infections, since reactivation of latent virus is thought to involve both signal transduction events and the induction of viral DNA synthesis. Thus, the expression and purification strategy outlined in this work provides an economical source of unmodified HSV-1 OBP that should prove useful in future in vitro studies. PMID- 11437605 TI - Purification and characterization of a Z-pro-prolinal-insensitive Z-Gly-Pro-7 amino-4-methyl coumarin-hydrolyzing peptidase from bovine serum--a new proline specific peptidase. AB - The study of a new proline-specific peptidase from bovine serum is presented. The enzyme readily cleaves the prolyl oligopeptidase (PO) substrate Z-Gly-Pro-MCA, liberating the fluorophore MCA, thus allowing quantification of enzyme activity. Unlike PO, however, this peptidase is completely insensitive to the PO-specific inhibitor Z-Pro-prolinal and has been designated Z-Pro-prolinal-insensitive Z-Gly Pro-MCA-hydrolyzing peptidase (ZIP). The two peptidases were successfully separated from each other by phenyl Sepharose hydrophobic interaction chromatography and the subsequent purification focused on the isolation of ZIP from bovine serum. In addition to phenyl Sepharose, calcium phosphate cellulose and DEAE anion-exchange chromatography were employed in the purification, with an overall enzyme yield of 33% and a purification factor of 4023. SDS-PAGE and size exclusion chromatography indicated a dimeric structure with a relative molecular mass of 174 kDa. The enzyme was stable over the pH range 2.5-10.0. Optimal activity was detected in the pH range 7.4-8.0. Isoelectric focusing revealed a pI of 5.68. Inhibition by AEBSF suggests the peptidase may be a serine protease and ZIP possibly contains a cysteine residue near the active site. alpha(2)M failed to inhibit activity, suggesting oligopeptidase specificity. HPLC analysis revealed a broad substrate specificity for proline-containing peptides. Kinetic analysis indicated that ZIP had a high affinity for Z-Gly-Pro-MCA with a K(m) of 54 microM deduced. Bovine serum ZIP exhibits biophysical characteristics both similar to and different from those of PO isolated from a number of sources and may serve an important physiological function in the degradation of bioactive oligopeptides. PMID- 11437606 TI - Purification of heterologous sarcoplasmic reticulum Ca2+-ATPase Serca1a allowing phosphoenzyme and Ca2+-affinity measurements. AB - We describe here a protocol to prepare milligrams of active and stable heterologous sarcoplasmic reticulum Ca(2+)-ATPase (Serca1a). Serca1a was tagged with 6 histidines at its C-terminal end and overexpressed using the baculovirus Sf9 system. In a first trial, Serca1a accounted for 24% of membrane proteins, 95% of which were inactive. Glucose in the culture medium reduced the production of Serca1a to 3 to 5% of membrane proteins and all Serca1a was active. Seventy-five percent of active Serca1a was solubilized by C(12)E(8) in the presence of phosphatidylcholine under conditions avoiding denaturation. Purification by Ni(2+)-nitrilo-triacetic acid affinity chromatography was tried, but only 3% of active Serca1a remained bound to the column, as if the His-tag were not accessible. Yields of 43% were reached by purification on reactive red 120 columns when eluting with 2 M NaCl. The purity was about 25% and Serca1a was stable for at least 1 week at 0 degrees C. Typically, 500 ml of culture medium produced 3 mg of active Serca1a and 1 mg of purified active Serca1a allowing measurements of phosphoenzyme (2 nmol/mg) or Ca(2+) affinity (2 microM at pH 7). PMID- 11437607 TI - High-level expression of three members of the murine angiogenin family in Escherichia coli and purification of the recombinant proteins. AB - Angiogenin (Ang) is a small basic protein which belongs to the pancreatic ribonuclease superfamily. It potently induces the formation of new blood vessels and has emerged as a promising anticancer target. Mice possess genes encoding one ortholog (mAng) and three homologs of Ang, designated angiogenin-related protein (mAngrp), angiogenin-3 (mAng-3), and angiogenin-4 (mAng-4). Structural and functional study of these homologs has been hampered by the low yield of protein from the existing heterologous expression system. In the experiments described, we used a pET expression vector to express these proteins in the cytoplasm of Escherichia coli BL21-CodonPlus(DE3)-RIL cells, whereupon substantial amounts of each accumulated in the form of insoluble aggregates. The proteins were renatured using an arginine-assisted procedure and subsequently purified by cation-exchange chromatography and reversed-phase HPLC; each purified protein was shown to be enzymatically active toward tRNA. The yields of pure mAngrp and mAng-3 were 7.6 and 12 mg/liter culture, respectively, representing substantial increases over previously reported experiments. This is also the first report of the expression and purification of mAng-4, obtained here in a yield of 30 mg/liter culture. The ready availability of milligram quantities of these proteins will enable further functional studies and high-resolution structural analyses to be conducted. PMID- 11437608 TI - NMR monitoring of accumulation and folding of 15N-labeled protein overexpressed in Pichia pastoris. AB - Postgenomic studies have led to an increasing demand for isotope-labeled proteins. We present a method for producing large quantities of truly native (15)N-labeled protein. Based on the secretion capabilities of the yeast Pichia pastoris, the recombinant protein is easily purified in a single step as it is secreted. Control of all nitrogen sources permits very high labeling yields. As a result, accumulation and folding of the recombinant protein can be monitored by heteronuclear NMR without purification. Comparison of sample spectra with the spectrum of the purified recombinant protein allows detection of the secreted protein in the culture and monitoring of its folding, from the start of the induction phase. The detection limit for a (15)N-labeled protein is estimated as 20 microM and corresponds, for a 10-kDa protein, to a load of 40 mg/liter in the fermentor. This concentration is reached by most reported preparations in P. pastoris. Further concentration by ultrafiltration would compensate for lower production. This procedure may be useful in many structural genomics and combinatorial chemistry screening projects where most protein productions meet the requirements for this method. PMID- 11437609 TI - Rapid two-step purification of a recombinant mouse Fab fragment expressed in Escherichia coli. AB - We report a rapid, large-scale process for the purification of a recombinant Fab fragment specific for the tobacco mosaic virus coat protein (Fab57P). The fragment is expressed periplasmically in Escherichia coli. The expression level was optimized in 0.3-L fermentors. The highest levels were obtained using the following conditions: (1) low postinduction temperature (21 degrees C), (2) combined use of two beta-lactam antibiotics (carbenicillin and ampicillin), (3) IPTG concentration 0.1 mM, (4) regulated pH 7.2, (5) 17-h induction time, and (6) conditions that reduce mechanical stress. Optimized large-scale fermentations were done in 15- and 300-L capacity fermentors. The recombinant Fab fragment was purified by two chromatographic steps. After disruption of the bacteria using an APV Gaulin homogenizer, the crude E. coli homogenate was directly applied, without centrifugation, to an SP Sepharose Big Beads column. The recombinant Fab fragment was eluted as a single peak in a sodium chloride gradient. The fragment was further purified by affinity adsorption to a column packed with Epoxy activated Sepharose 6B to which the antigen peptide NH(2)-CGS YNR GSF SQS SGLV CONH(2) had been coupled through its N-terminal cysteine. The purified Fab57P fragment showed one band in SDS-PAGE. The overall purification yield was 35%. PMID- 11437610 TI - Expression and purification of stable 33-kDa soluble human CD23 using the Drosophila S2 expression system. AB - CD23, a 45-kDa type II membrane glycoprotein present on B cells, monocytes, and other human immune cells, is a low-affinity receptor for IgE. The extracellular region of the membrane-bound human CD23 is processed into at least four soluble (s) CD23 forms, with apparent molecular masses of 37, 33, 29, and 25 kDa. High levels of sCD23 are found in patients with allergy, certain autoimmune diseases, or chronic lymphocytic leukemia. Therefore, inhibition of the processing of membrane-bound CD23 to control the cytokine-like effects of sCD23 offers a novel therapeutic opportunity. While the 37-, 29-, and 25-kDa forms of sCD23 have been expressed previously as recombinant proteins, the 33-kDa form has not been purified and characterized. To further investigate the multiple roles of sCD23 fragments and to devise assays to identify potent small-molecule inhibitors of CD23 processing, we have produced the 33-kDa form of sCD23 using Chinese hamster ovary (CHO) and Drosophila S2 cells. The CHO-expressed 33-kDa protein was found to undergo proteolytic degradation during cell growth and during storage of purified protein, resulting in accumulation of a 25-kDa form. The Drosophila system expressed the 33-kDa sCD23 in a stable form that was purified and demonstrated to be more active than the CHO-derived 25-kDa form in a monocyte TNFalpha release assay. PMID- 11437611 TI - Purification and characterization of human erythrocyte glucose transporter in decylmaltoside detergent solution. AB - The facilitative glucose transporter from human erythrocyte membrane, Glut1, was purified by a novel method. The nonionic detergent decylmaltoside was selected for solubilization on the basis of its efficiency to extract Glut1 from the erythrocyte membrane and its ability to maintain the protein in a monodisperse state. A positive, anion-exchange chromatography protocol produced a Glut1 preparation of 95% purity with little copurified lipid. This protein preparation exhibited cytochalasin B binding in detergent solution, as measured by tryptophan fluorescence quenching. The transporter existed as a monomer in decylmaltoside, with a Stokes radius of 50 A and a molecular mass of 147 kDa for the protein detergent complex. We screened detergent, pH, additive, and lipid and have found conditions to maintain Glut1 monodispersity for 8 days at 25 degrees C or over 5 weeks at 4 degrees C. This Glut1 preparation represents the best available material for two- and three-dimensional crystallization trials of the human glucose transporter protein. PMID- 11437612 TI - Expression of active human C1 inhibitor serpin domain in Escherichia coli. AB - Human C1 inhibitor is a highly glycosylated serine protease inhibitor of the serpin family. The protein contains two disulfide bonds. In this study, an N terminally truncated form of recombinant C1 inhibitor was overexpressed in Escherichia coli strains BL21(DE3) and AD494(DE3), the latter enabling the formation of disulfide bonds within the cytoplasm. With both strains, a major fraction of the recombinant protein produced appeared to be insoluble. However, the soluble fraction of lysates from strain AD494(DE3) inhibited the C1s target protease in functional assays. Recombinant C1 inhibitor produced in this strain also displayed the ability to complex with C1s in vitro. In contrast, lysates from strain BL21(DE3) displayed no C1 inhibitor activity. These data support the notion that glycosylation is not important, whereas disulfide bond formation appears to be essential for the production of an active recombinant C1 inhibitor. Thus, bacterial strains that permit the formation of disulfide bonds may represent a reliable system for the production of recombinant C1 inhibitor. However, a major obstacle to large-scale production will be to produce the protein in a soluble form. Attempts to increase the yield of soluble protein by coexpression of the GroEL/ES chaperonins resulted in an increase in solubility. PMID- 11437613 TI - SwellGel: an affinity chromatography technology for high-capacity and high throughput purification of recombinant-tagged proteins. AB - The revolution in genomics and proteomics is having a profound impact on drug discovery. Today's protein scientist demands a faster, easier, more reliable way to purify proteins. A high capacity, high-throughput new technology has been developed in Perbio Sciences for affinity protein purification. This technology utilizes selected chromatography media that are dehydrated to form uniform aggregates. The SwellGel aggregates will instantly rehydrate upon addition of the protein sample, allowing purification and direct performance of multiple assays in a variety of formats. SwellGel technology has greater stability and is easier to handle than standard wet chromatography resins. The microplate format of this technology provides high-capacity, high-throughput features, recovering milligram quantities of protein suitable for high-throughput screening or biophysical/structural studies. Data will be presented applying SwellGel technology to recombinant 6x His-tagged protein and glutathione-S-transferase (GST) fusion protein purification. PMID- 11437614 TI - The longitudinal course of adjustment after seizure surgery. AB - Surgical alleviation of chronic epilepsy can give rise to a process of adjustment as the chronically ill patient learns to become well. This process can manifest clinically as an array of symptoms which we have previously described as the 'burden of normality'. The aim of this study was to explore the longitudinal course of post-operative adjustment by mapping the incidence of symptoms of the burden of normality over a period of 2 years, and examining symptom occurrence relative to seizure outcome. A series of 90 anterior temporal lobectomy (ATL) patients was drawn from our Seizure Surgery Follow-up and Rehabilitation Program. All patients were prospectively assessed using the Austin CEP Interview, which covers symptoms of the burden of normality. In total, 66% of patients reported symptoms at some time within the first 2 years of surgery. Symptoms often emerged by the 3 month review, but were still seen frequently in the second year. At the 24 month review, patients who had been seizure free or experienced auras only within the previous 18 months were significantly more likely to report symptoms compared to patients who had experienced complex partial and/or generalized tonic clonic seizures (P = 0.03). Surgical alleviation of seizures in chronic epilepsy brings with it the burden of normality. Recognition of this syndrome is essential in maximizing patient outcome. PMID- 11437615 TI - Theoretical and practical considerations in the psychological and educational assessment of the student with intractable epilepsy: dynamic assessment as an adjunct to static assessment. AB - Assessing the student with intractable epilepsy requires skill not only in evaluating cognitive problems, but also detecting seizures and discovering how to adapt instruction to minimize their negative impact on learning. Ironically, assessment efforts are seen as compromised by the occurrence of seizures during testing, when determining how seizure events may interfere with learning and the instructional modifications that are necessary to cope with them, should be a key part of assessment. A dual approach to assessment is recommended that combines the identification of cognitive deficits with an evaluation of how recurring seizures may prevent the student from engaging in instruction. Without also evaluating the student's response to instruction, teaching to specific cognitive needs is limited by insufficient knowledge about how to keep the student involved in instruction when seizures occur. Static assessment evaluates cognitive functioning at the time of testing, without changing the way that the student learns and responds. By engaging the student in teaching/learning sessions, dynamic assessment explores how the student best learns despite cognitive deficits and the disruptive effect of seizures. This paper includes a description of the authors'experience in using dynamic assessment as an adjunct to static assessment in evaluating a student with intractable epilepsy. PMID- 11437616 TI - Adjustment and coping in epilepsy. AB - Our study investigated interrelationships between problems in psychosocial adjustment, coping and epilepsy variables. Establishing the cross-cultural applicability of the Washington Psychosocial Seizure Inventory (WPSI) was an additional objective. The WPSI, Ways of Coping Scale, Modified Version, as well as scales measuring depression and anxiety were administered to 310 outpatients with epilepsy. When the scores of patients with high Lie scores were eliminated, the WPSI profiles were found to be similar to former studies, with some score elevations in emotional adjustment and interpersonal adjustment. When relationships between adjustment variables and coping were modelled, coping was found to be a mediator between the effects of interpersonal and emotional adjustment and integration to the broader social context (vocational adjustment). Family background was found to be a significant predictor of the emotional well being and interpersonal adjustment scores of the patients. Results support the central role of coping and emotional well-being and emphasize the importance of family factors in adjustment to epilepsy. Psychotherapy and psychological interventions could support coping with illness, primarily through elimination of negative family and social effects and treatment of emotional problems. PMID- 11437617 TI - A study of the relationship between metabolism using 1H-MRS and function using several neuropsychological tests in temporal lobe epilepsy. AB - Several investigators have reported on the relationship between metabolism, using magnetic resonance spectroscopy (MRS), and function, using neuropsychological tests in temporal lobe epilepsy (TLE) patients, but the opinions regarding the results remain in contention. The aim of this study is to examine the relationship between metabolism, using proton MRS ((1)H-MRS), and function using several neuropsychological tests in the temporal lobes of TLE patients. We studied 29 TLE patients at our hospital using(1)H-MRS and neuropsychological tests. We used a clinical 1.5 T MR unit. We conducted five neuropsychological tests to examine the function of the left or right temporal lobe. There were significant correlations between the N-acetylaspartate/creatine + phosphocreatine (NAA/Cr) ratios and the scores of almost all of the neuropsychological tests for the temporal lobe function ipsilateral to the spike focus. However, in two Wechsler Memory Scale-Revised (WMS-R) subtests we found no significant correlation in the ipsilateral side. These findings suggest that the NAA/Cr ratios, which reflect neural metabolism, are closely related to function in the temporal lobes of TLE patients. The disparity between the results in two subtests of WMS-R show that several tests may be necessary in order to assess temporal lobe function. PMID- 11437618 TI - A pilot study on benign partial epilepsy in children with complex partial seizures. AB - In a review of 145 children with partial onset epilepsy, the authors were able to determine a focus of children whose complex partial seizures (CPS) ran a benign course, who had no identifiable lesion on scanning and whose EEG focus was not fixed, but tended to shift. The authors suggest that a benign form of CPS in children can be recognised. PMID- 11437619 TI - Long-term follow-up study of Lennox-Gastaut syndrome in patients with severe motor and intellectual disabilities: with special reference to the problem of dysphagia. AB - A long-term follow-up study of Lennox-Gastaut syndrome (LGS) ( > 10 years) was conducted with 38 patients with severe motor and intellectual disabilities (SMID) to clarify the relationship between the rapid development of dysphagia and epileptic seizures, and to elucidate the long-term evolution of LGS in patients with SMID. Those who showed a relatively favourable seizure outcome were compared to those with a poor seizure outcome. Poor seizure outcome correlated strongly with: (a) an early appearance of dysphagia and additional deterioration of the already retarded mental function; (b) a predominance of atypical absence seizures; and (c) persistent frequent epileptiform discharges during electroencephalographic evaluations. Neither age at seizure onset nor intelligence level at the time of the last examination was correlated with seizure prognosis. Further, seizure prognosis was not related to the aetiology of LGS. Repeated seizures apparently caused development of progressive epileptic encephalopathy, in addition to the underlying severe brain damage. Since development of dysphagia burdens an already severely handicapped patient with intensive medical care, efforts to reduce the seizures and design a long-term care plan are of great importance. PMID- 11437620 TI - The attitude of courts in England to compensation for post-traumatic epilepsy. AB - The attitudes of courts in England to the assessment of damages for post traumatic epilepsy have dramatically changed over the last 20-30 years. In assessing damages for post-traumatic epilepsy the courts are faced with a number of considerations: epilepsy can appear several years after the injury; epilepsy is not a homogeneous condition; the eventual prognosis is unknown; the epilepsy may not have been directly due to the trauma; and epilepsy affects life expectancy and employment. Damages were originally fixed at the point of compensation, and these rather crude calculations led to both over- and under compensation. This situation was improved in 1985, when courts were permitted to award damages on the assumption that epilepsy would not occur or worsen, and further damages should these assumptions prove to be incorrect. The courts in England still depend, however, upon the evidence of expert witnesses chosen by the plaintiff and defendant. A tension thus exists between the duty of expert witnesses to the court and the understandable inclination of expert witnesses to support the party that has instructed them. The Woolf report has led to changes in the responsibilities of expert witnesses, and will hopefully remedy many of the inconsistencies and inequities that occur. PMID- 11437621 TI - Psychogenic seizures mimicking juvenile myoclonic epilepsy: case reports. AB - We describe two patients with psychogenic seizures of rare semiology. Both patients (a 23-year-old man and a 26-year-old woman) attended the emergency department because status epilepticus with myoclonic seizures had been diagnosed. Seizures were documented with video-electroencephalography. Semiology of seizures were brief myoclonia of both arms resulting in a short elevation of both arms without impairment of consciousness. Ictal EEG registration was without abnormal finding. Psychiatric diagnostic assessment suggested a dissociative disorder and mild depression, respectively. During psychiatric treatment seizures occurred only rarely within a 3-5 months follow-up. One should be aware that juvenile myoclonic epilepsy may be mimicked by psychogenic seizures PMID- 11437622 TI - CPD-Education and self-assessment: Epilepsy and pregnancy. AB - Pregnancies in women with epilepsy are high risk and need careful management by both the medical and obstetric teams due to the increased incidence of complications and adverse outcomes of pregnancy. By the time a pregnant woman with epilepsy presents, the foetus is virtually fully formed and the opportunity for altering drug treatment has passed. Women need to be counselled and told to seek advice about their anticonvulsant therapy should they wish to become pregnant. All major anticonvulsant drugs are teratogenic but the main risk to the developing foetus appears to be when the mother is on polytherapy especially if sodium valproate forms part of the combination. Folate supplements (5 mg) before conception are advisable. There appears to be a minor but significant increased risk of maternal complications in women with epilepsy such as hyperemesis gravidarum, pre-eclampsia and eclampsia, vaginal bleeding and premature labour. In the majority of women seizure control will not alter during pregnancy. Oral vitamin K should be given to the mother receiving enzyme-inducing antiepileptic drugs. Post-natal infant development: there is an increased risk of prematurity (9-11%), stillbirth, neonatal and perinatal death, haemorrhagic disease of the newborn, low Apgar scores and low birth weight (7-10%). Breast feeding: virtually all the anticonvulsant drugs are excreted in breast milk in low concentrations. Feeding difficulties, irritability and lethargy can occur. However, the benefits of breast feeding usually far outweigh any minor risks to the baby. PMID- 11437623 TI - Epilepsy and the ovary (cutting out the hysteria). AB - This review touches on the historical links between epilepsy, seizures and the uterus and ovaries which have fascinated and misled physicians since Greco-Roman times. It then examines present knowledge of ovarian function and its effect on epileptic activity and vice versa before exploring the modern controversy about polycystic ovaries and the polycystic ovary syndrome, epilepsy and anticonvulsant medication. Based on present evidence, women with epilepsy are more prone to develop polycystic (polyfollicular) ovaries than other women due to the epilepsy itself. But women with epilepsy related polycystic (polyfollicular) ovaries are vulnerable to the effects of sodium valproate (possibly particularly during adolescence) and may develop the polycystic ovary syndrome: this is reversible if the valproate is withdrawn. Lamotrigine and carbamazepine seem to prevent the development of the syndrome. PMID- 11437624 TI - Case reports. PMID- 11437625 TI - The 'number needed to treat' with Levetiracetam (LEV): comparison with the other new antiepileptic drugs (AEDs). PMID- 11437628 TI - Extinguishing maternal immune responses during pregnancy: implications for immunosuppression. AB - Mammals owe their existence to immunosuppressive processes that prevent fetal rejection in utero. Blocking tryptophan catabolism during murine pregnancy allows maternal T cells to provoke fetal allograft rejection. Cells expressing indoleamine 2,3-dioxygenase (IDO), which catabolizes tryptophan, prevent T cell cycle progression and enhance activation induced T cell death. Here, we discuss the role of cells expressing IDO in regulating maternal T cell immunity during pregnancy and consider whether this mechanism might contribute to immunological discrimination by promoting T cell tolerance in other circumstances. PMID- 11437629 TI - Pregnancy: success and failure within the Th1/Th2/Th3 paradigm. AB - Evidence from studies on murine and human pregnancy points to a strong association between maternal Th2-type immunity and successful pregnancy on the one hand and between Th1-type immune reactivity and pregnancy loss on the other. While there is a paucity of data from human pregnancy indicating that Th1-type immune effectors actually lead to pregnancy, it is difficult to ignore the compelling evidence linking inappropriate Th1-type immunity to pregnancy loss. Th2-type immunity and TGF beta secreted by Th3 cells may play protective roles during pregnancy, hence the nexus between a Th2/Th3 shift and successful pregnancy. This paper examines these associations and discusses possible mechanisms underlying immunologically mediated pregnancy failure. PMID- 11437630 TI - The role of gamma/delta T cells in the feto-maternal relationship. AB - Polymorphic MHC is absent from the trophoblast, therefore, it resists NK as well as CTL-mediated lysis in vitro. Activated gamma / delta TCR positive cells are significantly enriched in the decidua as well as in peripheral blood of healthy pregnant women. Human peripheral gamma / delta lymphocytes preferentially express the V gamma 9/V delta 2 TCR, whereas those of the decidua use the V delta 1 chain. These subpopulations are functionally polarized, the former being Th1, the latter Th2. Potentially cytotoxic V delta 2+ lymphocytes recognize HLA-E on the trophoblast via the CD94/NKG2A receptor, which induces an inhibitory signal, thus potentially inhibiting Th1 type cytokine production. PMID- 11437631 TI - Functions of uterine natural killer cells are mediated by interferon gamma production during murine pregnancy. AB - The dominant lymphocytes in healthy human and murine implantation sites are pregnancy-associated uterine natural killer (uNK) cells. These cells produce 90% of pregnancy-induced, uterine interferon (IFN)- gamma, a cytokine that regulates expression of more than 0.5% of the mouse genome. Implantation sites in uNK cell deficient and IFN- gamma -signal-disrupted mice display anomalies in decidua and its spiral arteries. Reconstitution of uNK cell-deficient females with bone marrow containing normal NK cell progenitors, establishes uNK cells and reverses the anomalies. Grafts from IFN- gamma(-/-)mice are restored uNK cells, but the uNK cells did not reverse the phenotypes. This review focuses on the functions of uNK cell-derived IFN- gamma and the genes that it may regulate in the pregnant uterus. PMID- 11437632 TI - The role of semen in induction of maternal immune tolerance to pregnancy. AB - Successful pregnancy requires a state of maternal immune 'tolerance' to accommodate antigens expressed by the conceptus. Implantation failure and placental pathologies largely reflect insufficiencies in maternal immune adaptation, but progress in devising therapeutic strategies to treat these conditions is stalled because the mechanisms underlying the induction and maintenance of maternal tolerance are unknown. Increasingly, clinical and experimental data support the proposal that insemination has consequences for the reproductive process beyond delivery of male gametes. An emerging hypothesis, based mainly on clinical observations and experiments in mice, is that insemination is causally linked to the activation and expansion of populations of lymphocytes mediating forms of 'active' immune tolerance in the implantation site. This review examines existing evidence for a role for semen in the immunology of pregnancy, highlighting the limitations of our existing knowledge and the prospects for future research and its clinical application. PMID- 11437633 TI - Procoagulants in fetus rejection: the role of the OX-2 (CD200) tolerance signal. AB - The spontaneous loss of normal karyotype embryos may be initiated or prevented by the maternal immune system. In mice, loss between the time of implantation (day 4.5) and formation of a vascularized placenta (day 9.5) when the embryo is too large to survive by diffusion alone, is analogous to occult pregnancy failure in humans. They are called occult because usually the woman does not know she is pregnant. From studies in mice, these early losses have a different mechanism than abortion of a vascularized placenta (analogous to clinically evident human spontaneous miscarriage). The latter depend on the activation of the novel prothrombinase fgl2 on the fetal trophoblast and in maternal decidua by the T helper-1 (Th1) type cytokines TNF- alpha+gamma -interferon that arise from NK cells and NK gammadelta T cells; conversion of prothrombin to thrombin which in turn generates IL8 that activates polymorphonuclear leukocytes leads to embryonic death. These inflammatory processes are counteracted by Th2/3-type cytokines that arise in part from V gamma 1 delta 6 T cells reacting to, as yet, unidentified trophoblast antigens in the presence of the 'tolerance signaling molecule' OX-2. By contrast, peri-implantation losses (between implantation and formation of a vascularized placenta, analogous to occult losses in humans) appear to be dependent upon perforin(+)cells, complement activation, and products of alphabeta T and NK alphabeta T cells, but not on TNF- alpha or procoagulant activation. Similarities and differences between findings in the mouse and human, and the potential evolutionary significance of mechanisms affecting reproductive success are reviewed. PMID- 11437634 TI - Retinol potentiates acetaminophen-induced hepatotoxicity in the mouse: mechanistic studies. AB - This study was designed to elucidate the mechanism of retinol's potentiation of acetaminophen-induced hepatotoxicity. To accomplish this, the major bioactivation and detoxification pathways for acetaminophen were investigated following retinol (75 mg/kg/day, 4 days), acetaminophen (400 mg/kg), and retinol + acetaminophen treatment. Hepatic microsomes were used to determine the catalytic activity and polypeptide levels of cytochrome P450 enzymes involved in the murine metabolism of acetaminophen. Results showed that the catalytic activity and polypeptide levels of CYP1A2, CYP2E1, and CYP3A were unchanged in the treatment groups compared to vehicle and untreated controls. In combination, retinol + acetaminophen caused a significantly greater depletion of GSH compared to corn oil + acetaminophen (0.36 +/- 0.11 vs 0.89 +/- 0.19 micromol/g, respectively, p < 0.05). This greater GSH depletion correlated with a higher degree of hepatic injury in the retinol + acetaminophen-treated animals but is probably not the cause of the potentiated injury since the results showed that retinol treatment itself did not alter hepatic glutathione (3.34 +/- 0.43 vs 3.44 +/- 0.46 micromol/g for retinol vs vehicle, respectively). However, hepatic UDPGA stores were decreased in the retinol-treated group compared to untreated and corn oil controls (54.6 +/- 10.6 vs 200.6 +/- 17.6 nmol/g for retinol and untreated control, respectively, p < 0.001). This demonstrates that there is significantly less hepatic UDPGA available for conjugation following retinol administration. The results suggest that decreased hepatic UDPGA is likely the cause of retinol's potentiation of acetaminophen-induced hepatic injury. PMID- 11437635 TI - Longitudinal distribution of ozone and chlorine in the human respiratory tract: simulation of nasal and oral breathing with the single-path diffusion model. AB - In the single-path model of the respiratory system, gas transport occurs within a conduit of progressively increasing cross-sectional and surface areas by a combination of flow, longitudinal dispersion, and lateral absorption. The purpose of this study was to use bolus inhalation data previously obtained for chlorine (Cl(2)) and for ozone (O(3)) to test the predictive capability of the single-path model and to adjust input parameters for applying the model to other exposure conditions. The data, consisting of uptake fraction as a function of bolus penetration volume, were recorded on 10 healthy nonsmokers breathing orally as well as nasally at alternative air flows of 150, 250, and 1000 ml/s. By employing published data for airway anatomy, gas-phase dispersion coefficients, and gas phase mass transfer coefficients while neglecting diffusion limitations in the mucus phase, the single-path model was capable of predicting the uptake distribution for O(3) but not the steeper distribution that was observed for Cl(2). To simultaneously explain the data for these two gases, it was necessary to increase gas-phase mass transfer coefficients and to include a finite diffusion resistance of O(3) within the mucous layer. The O(3) reaction rate constants that accounted for this diffusion resistance, 2 x 10(6) s(-1) in the mouth and 8 x 10(6) s(-1) in the nose and lower airways, were much greater than previously reported reactivities of individual substrates found in mucus. PMID- 11437636 TI - Zinc-induced activation of the human cytomegalovirus major immediate-early promoter is mediated by metallothionein and nuclear factor-kappaB. AB - We previously reported that major immediate-early promoter (MIEP) activity was regulated by intercellular zinc levels. In this report, we elucidate the mechanisms involved in this phenomenon. In luciferase reporter assays, zinc induced activation of MIEP (-735/+62) was decreased with deletion of the promoter in stages, and MIEP (-117/+62) did not respond to zinc. The time course of the activity of MIEP responding to diethylenetriamine pentaacetic acid and zinc was not parallel with metallothionein (MT) promoter, which contains metal responsive elements. SV40 promoter that contains AP-1 binding sites, a candidate for the zinc-responsive motif in the MIEP, was not affected by zinc under our conditions. The activation of MIEP (-735/+62) by zinc was prevented with NF-kappaB decoy. When three kappaB motifs from the enhancer in the MIEP were inserted in the front of the zinc-nonresponsive MIEP (-117/+62), it became responsive to zinc. Moreover, overexpression of MT up-regulates the DNA binding of NF-kappaB and NF kappaB-induced activation of transcription. These findings strongly suggest that MT and NF-kappaB act as mediator/regulator in zinc-induced activation of MIEP. PMID- 11437637 TI - Accelerated DNA adduct formation in the lung of the Nrf2 knockout mouse exposed to diesel exhaust. AB - Diesel exhaust (DE) has been recognized as a noxious mutagen and/or carcinogen, because its components can form DNA adducts. Mechanisms governing the susceptibility to DE and the efficiency of such DNA adduct formation require clarification. The transcription factor Nrf2 is essential for inducible and/or constitutive expression of a group of detoxification and antioxidant enzymes, and we hypothesized that the nrf2 gene knockout mouse might serve as an excellent model system for analyzing DE toxicity. To address this hypothesis, lungs from nrf2(-/-) and nrf2(+/-) mice were examined for the production of xenobiotic-DNA adducts after exposure to DE (3 mg/m(3) suspended particulate matter) for 4 weeks. Whereas the relative adduct levels (RAL) were significantly increased in the lungs of both nrf2(+/-) and nrf2(-/-) mice upon exposure to DE, the increase of RAL in the lungs from nrf2(-/-) mice exposed to DE were approximately 2.3-fold higher than that of nrf2(+/-) mice exposed to DE. In contrast, cytochrome P4501A1 mRNA levels in the nrf2(-/-) mouse lungs were similar to those in the nrf2(+/-) mouse lungs even after exposure to DE, suggesting that suppressed activity of phase II drug-metabolizing enzymes is important in giving rise to the increased level of DNA adducts in the Nrf2-null mutant mouse subjected to DE. Importantly, severe hyperplasia and accumulation of the oxidative DNA adduct 8 hydroxydeoxyguanosine were observed in the bronchial epidermis of nrf2(-/-) mice following DE exposure. These results demonstrate the increased susceptibility of the nrf2 germ line mutant mouse to DE exposure and indicate the nrf2 gene knockout mouse may represent a valuable model for the assessment of respiratory DE toxicity. PMID- 11437638 TI - A physiologically based pharmacokinetic model for ethylene oxide in mouse, rat, and human. AB - Ethylene oxide (EO) is widely used as a gaseous sterilant and industrial intermediate and is a direct-acting mutagen and carcinogen. The objective of these studies was to develop physiologically based pharmacokinetic (PB-PK) models for EO to describe the exposure-tissue dose relationship in rodents and humans. We previously reported results describing in vitro and in vivo kinetics of EO metabolism in male and female F344 rats and B6C3F1 mice. These studies were extended by determining the kinetics of EO metabolism in human liver cytosol and microsomes. The results indicate enzymatically catalyzed GSH conjugation via cytosolic glutathione S-transferase (cGST) and hydrolysis via microsomal epoxide hydrolase (mEH) occur in both rodents and humans. The in vitro kinetic constants were scaled to account for cytosolic (cGST) and microsomal (mEH) protein content and incorporated into PB-PK descriptions for mouse, rat, and human. Flow-limited models adequately predicted blood and tissue EO levels, disposition, and elimination kinetics determined experimentally in rats and mice, with the exception of testis concentrations, which were overestimated. Incorporation of a diffusion-limited description for testis improved the ability of the model to describe testis concentrations. The model accounted for nonlinear increases in blood and tissue concentrations that occur in mice on exposure to EO concentrations greater than 200 ppm. Species differences are predicted in the metabolism and exposure-dose relationship, with a nonlinear relationship observed in the mouse as a result of GSH depletion. These models represent an essential step in developing a mechanistically based EO exposure-dose-response description for estimating human risk from exposure to EO. PMID- 11437639 TI - Mercuric ion attenuates nuclear factor-kappaB activation and DNA binding in normal rat kidney epithelial cells: implications for mercury-induced nephrotoxicity. AB - Mercuric ion (Hg(2+)), one of the strongest thiol-binding agents known, mediates the toxicity associated with elemental, inorganic, and organic mercurial compounds. Studies of cellular events associated with Hg(2+) toxicity have focused largely on disruption of cell membranes and impairment of mitochondrial functions. In contrast, few studies have sought to define the specific molecular mechanisms through which Hg(2+) might affect toxicity via alteration of thiol dependent signal transduction pathways that regulate cell proliferation and survival. Of particular interest in this regard is the effect of Hg(2+) on nuclear factor-kappaB (NF-kappaB), a pleiotropic transcriptional factor that is known to require reduced cysteine moieties at critical steps of activation and DNA binding. Here, we evaluated the effects of Hg(2+) on the expression of NF kappaB in normal rat kidney epithelial (NRK52E) cells, a principal target of Hg(2+) toxicity. The lipopolysaccharide (LPS)-inducible form of NF-kappaB was readily detected in kidney cells and has been characterized as the p50p65 heterodimer. NF-kappaB-DNA binding was prevented in a dose-related manner by Hg(2+) (0-55 microM) in vitro when added to DNA binding reactions containing the nonthiol reducing agent Tris(2-carboxyethyl)phosphine hydrochloride (TCEP). Similarly, Hg(2+) at the same concentrations prevented DNA binding of a human recombinant wild-type p50p50 homodimer in binding reactions, and this effect was attenuated using a mutant form of the p50 protein containing a cys(62)-->ser(62) mutation. The inhibition of p50-DNA binding by Hg(2+) was reversible in a dose related manner in vitro by competitive thiols DTT, GSH, and l-cysteine in binding reactions. In contrast, competitive thiols added to nuclear binding reactions were unable to reverse attenuation of LPS-mediated NF-kappaB-DNA binding affinity when cells were pretreated in vivo with Hg(2+) at concentrations as low as 2 microM prior to LPS administration. Immunoblot analyses indicted that Hg(2+) pretreatment of kidney cells substantially diminished, in a dose-related manner, the concentration of p65 translocated into the nucleus following LPS administration. Additionally, Hg(2+) pretreatment impaired both the phosphorylation and degradation of IkappaBalpha, suggesting a specific effect on NF-kappaB activation at the level of IkappaBalpha proteolysis. Finally, Hg(2+) at concentrations as low as 5 microM significantly diminished NF-kappaB-mediated transcriptional activity when administered to kidney cells transiently transfected with an NF-kappaB-driven luciferase reporter gene (pLuc-4xNF-kappaB) prior to LPS treatment. These findings demonstrate that Hg(2+), at low cellular concentrations, attenuates NF-kappaB activation at sites associated with IkappaBalpha phosphorylation and degradation, nuclear translocation of the p50p65 heterodimer, and association of p50-cys(62) with the DNA kappaB binding site. Attenuation of NF-kappaB activation by Hg(2+) through these mechanisms may underlie apoptotic or other cytotoxic responses that are known to be associated with low level Hg(2+) exposure in kidney epithelial cells. PMID- 11437640 TI - Re: ethylbenzene induces microsomal oxygen free radical generation: antibody directed characterization of the responsible cytochrome p450 enzymes (toxicol. appl. pharmacol. 164, 305-311, 2000). PMID- 11437643 TI - Effects of triphenyltin on cytosolic Na(+) and Ca(2+) response to glucose and acetylcholine in pancreatic beta-cells from hamster. AB - Oral administration of triphenyltin chloride (TPT) (60 mg/kg body wt) inhibits insulin secretion by perturbing the cytoplasmic Ca(2+) concentration ([Ca(2+)]i) in pancreatic beta-cells of the hamster. To test the possibility that the abnormal levels of [Ca(2+)]i induced by TPT administration could be due to a defect in the cytoplasmic Na(+) concentration ([Na(+)]i) in the beta-cells, we investigated the effects of TPT administration on the changes of [Na(+)]i and [Ca(2+)]i induced by glucose or acetylcholine (ACh) and on the [Na(+)]i induced by ouabain, a potent inhibitor of Na(+),K(+)-ATPase. The changes of [Na(+)]i and [Ca(2+)]i were measured in islet cells loaded with sodium-binding benzofuran isophthalate and fura 2, respectively. TPT administration strongly reduced the rise in [Ca(2+)]i induced by 15 mM glucose with and without extracellular 135 mM Na(+). TPT administration also significantly reduced the rise of [Ca(2+)]i by 100 microM ACh in the presence of 5.5 or 15 mM glucose but not the amplitude of [Ca(2+)]i by 100 microM ACh in Na(+)-free medium. TPT administration attenuated the rise in [Na(+)]i induced by 100 microM ACh in the presence of either 3 or 5.5 mM glucose. However, TPT administration did not impair the [Na(+)]i in the presence of glucose (3, 5.5, and 15 mM) or of 100 microM ouabain with 3 mM glucose. TPT administration significantly suppressed the insulin secretion induced by 15 and 27.8 mM glucose or 100 microM ACh in the presence of 5.5 mM glucose. Our study suggests that triphenyltin has inhibitory effects on the cellular Ca(2+) response due to a reduction of [Ca(2+)]i after Na(+)-dependent and Na(+)-independent depolarization in islet cells of the hamster. PMID- 11437644 TI - Cell surface regulation of silica-induced apoptosis by the SR-A scavenger receptor in a murine lung macrophage cell line (MH-S). AB - Scavenger receptors (SR) are responsible for recognition of ligands as diverse as oxidized LDL (endogenous) to respirable particulates (exogenous). A number of recent studies have suggested that these SR ligands induce apoptosis of macrophages. However, the mechanism by which SR triggers apoptosis is not understood. This study used a murine alveolar macrophage cell line (MH-S) to investigate the role of the SR in caspase activation. The presence of SR on MH-S cells was confirmed by FACS analysis and was similar to the distribution found on murine alveolar macrophages. The activity of caspases 1, 3, and 6 was measured following a 6-h exposure to crystalline silica with and without blockers of the SR. Caspase activities were determined by hydrolysis of specific chromogenic substrates and formation of an active enzymatic form (Western for active caspase 3). Silica stimulated significant caspase activity, apoptosis, and necrosis of MH S cells, which was attenuated by 2F8 (a blocking antibody) and polyinosinic acid (a nonspecific SR antagonist). The results indicate that the SR are necessary for caspase activation and subsequent apoptosis (as well as necrosis) caused by silica in macrophage cells. PMID- 11437645 TI - Seriate histomorphometry of whole rat stomach: an accurate and reliable method for quantitative analysis of mucosal damage. AB - The evaluation of mucosal damage in experimental models of gastric injury is commonly based on macroscopic detection of gross lesions and/or histological examination of tissue samples and is limited by the subjectivity of the examiner and by the paucity of nonrepresentative samples. This study proposes a novel method for the histomorphometric analysis of gastric damage, based on the examination of seriate parallel strips taken from whole rat stomachs. Strips were cut perpendicular to the lesser curvature, placed on a glass slide, with the side of each strip distal to the pylorus upward, and processed for routine histology. Sections were then observed by light microscopy: the length of damaged mucosa divided by the total length of mucosa, measured on a micrometric scale and expressed in percentage values, was indicated as the lesion index. Furthermore, to evaluate the severity of the damage, three types of lesions were discriminated depending on their depth: type I, lysis of luminal cells; type II, damage involving the cells lying on both surface mucosa and gastric pits; and type III, damage involving the lower part of the lamina propria with injury of glands associated with detachment of whole mucosal layers. Three models of acute gastric damage (ethanol, hemorrhagic shock, and indomethacin) were examined and treatment was also carried out with the antiulcer drugs omeprazole, ranitidine, and misoprostol, to show the advantages of this histomorphometric approach. The results indicate that this method allows an accurate quantitative analysis of gastric damage, and the effects of different antiulcer drugs can be better discriminated. PMID- 11437646 TI - Cisplatin ototoxicity: involvement of iron and enhanced formation of superoxide anion radicals. AB - Since there are indications that iron influences cisplatin nephrotoxicity, we studied the role of iron in cisplatin ototoxicity in an in vitro model of the neurosensory epithelium of the guinea pig cochlea. Viability tests showed that Deiters and Hensen cells were not damaged and inner hair cells were only slightly damaged by cisplatin (50 microM). The outer hair cells were most sensitive to cisplatin toxicity. The iron chelator 2,2'-dipyridyl provided partial protection against cisplatin-induced cell death. In addition, we studied the influence of the iron chelators 2,2'-dipyridyl and deferoxamine on the chelatable iron pool in the various cells of the neurosensory epithelium using the fluorescent iron indicator Phen Green SK. Both chelators decreased the chelatable iron accessible to Phen Green SK, although the effect of deferoxamine was weaker because it entered the cells more slowly. The cellular concentration of the chelatable iron was measured using Phen Green SK and quantitative laser scanning microscopy. The concentration of chelatable iron in the inner ear cells ranged from 1.3 +/- 0.4 microM iron in inner hair cells to 3.7 +/- 1.7 microM iron in Hensen cells and did not correlate with the various cell types' susceptibility to cisplatin. Furthermore, cisplatin did not raise the intracellular chelatable iron concentration but enhanced the production of superoxide anions inside the neurosensory epithelium, especially inside the hair cells, as detected by the nitrotetrazolium blue reduction assay. Our conclusion is that cisplatin ototoxicity is partially mediated by an iron-dependent pathway and is associated with an enhanced formation of superoxide anions. PMID- 11437647 TI - Involvement of Bcl-2 family genes and Fas signaling system in primary and secondary male germ cell apoptosis induced by 2-bromopropane in rat. AB - Epidemiological surveys and animal experimental studies suggest that exposure to 2-bromopropane (2-BP) could result in reproductive and hematopoietic disorders. The objectives of this study were to investigate the role of apoptosis in 2-BP induced testicular toxicity and whether this process involves Bcl-2 family genes and the Fas signaling system. Rats were injected percutaneously with 1350 mg/kg 2 BP for 1 to 5 days and then were euthanized at 6 or 12 h after one dose, 6 h after two, three, or five doses, and 2 or 9 days after the final treatment. Light and electron microscopic analyses, TUNEL staining of DNA fragments, agarose gel electrophoresis of low-molecular-weight DNA, and Western blotting analysis of Bcl 2 family proteins and Fas receptor and ligand were conducted. Two-day treatment resulted in selective degeneration of spermatogonia with marked nuclear chromatin condensation. DNA ladder formation on the agarose gel further validated the findings of TUNEL-stained apoptotic cells. The percentage of apoptotic-positive tubules and apoptotic cell index increased time dependently. 2-BP treatment resulted in two distinct morphological changes: an immediate effect on spermatogonia and secondary apoptosis of spermatocytes 9 days after treatment. Downregulation of Bcl-2 after the first or second injection of 2-BP and upregulation of Bax after the first treatment contributed to the initiation of primary apoptosis of spermatogonia. Expression of FasL was inhibited while expression of Fas increased after the 2-BP treatment and remained at levels about two times of the control. However, it increased about sixfold of the control by day 9 after final injection, which contributed to the induction of secondary apoptosis of spermatocytes. Our results indicate that 2-BP resulted in apoptotic death of testicular germ cells and that this process involves the Bcl-2 family genes and the Fas signaling system. PMID- 11437648 TI - Inhibition of axonal morphogenesis by nonlethal, submicromolar concentrations of methylmercury. AB - We investigated the effects of sublethal concentrations of the neurotoxicant methylmercury (MeHg) on the developmental progression of cultured neurons to the stage of axonal morphogenesis. Chick (E8) forebrain neurons in vitro develop axons by a stereotyped developmental sequence nearly identical to that of widely used rat hippocampal neurons, but at much less cost and difficulty. In this chick forebrain system, 40% of neurons develop long axons after 2 days in culture, and 80% have axons after 4 days. A single, 2-h exposure to 0.5 or 0.25 microM MeHg reduced the number of neurons developing axons to approximately half that of controls without causing significant cell death for at least 2 days after treatment. Although MeHg caused an immediate depolymerization of neuronal microtubules, after 1 day of recovery the microtubule array of MeHg-treated neurons was indistinguishable by immunofluorescent assay from that of untreated cells at equivalent development stages. Thus, the inhibition of axonal development by submicromolar concentrations of MeHg did not appear to be the direct effect of microtubule disassembly. Chelation of Ca(2+) during MeHg exposure appeared to exert a small immediate protective effect, as previously reported, but was itself toxic within 1 day after chelation. We suggest that this inhibition of axonal morphogenesis by acute, sublethal concentrations of MeHg may play a role in the developmental syndrome caused by environmental exposure to MeHg. PMID- 11437649 TI - An alternative promoter contributes to tissue- and inducer-specific expression of the rat UDP-glucuronosyltransferase 1A6 gene. AB - UDP-glucuronosyltransferase 1A6 (UGT1A6), a key enzyme catalyzing the glucuronidation of small planar phenols and amines, is expressed in a tissue- and inducer-dependent manner. Expression is high in kidney, gastrointestinal tract, and induced liver, with low expression in spleen, lung, and ovary. Exposure to certain chemicals, such as 3-methylcholanthrene, benzo[a]pyrene, beta naphthoflavone, and oltipraz elevates UGT1A6 mRNA in liver and to a lesser extent gastrointestinal tract and kidney, but not in other tissues. The mechanisms underlying this complex pattern of expression have been elusive. We have identified a new type of UGT1A6 mRNA (class 2) that differs in its 5' untranslated sequence. The class 2 transcript is the more abundant type expressed in liver, gastrointestinal tract, and kidney. Transcription of the class 2 mRNA is initiated 107 bases 5' of the UGT1A6 coding exon. The promoter region flanking the transcription start site contains an HNF1-like binding site identical to that in the human UGT1A6 gene. Both class 1 and class 2 mRNAs were elevated in liver by 3-methylcholanthrene, benzo[a]pyrene, beta-naphthoflavone, and oltipraz, with preferential elevation of class 1 occurring after 3-methylcholanthrene and benzo[a]pyrene treatment. These data suggest that transcription from a second promoter contributes to tissue- and inducer-specific expression of rat UGT1A6. PMID- 11437650 TI - Fumonisin-induced tumor necrosis factor-alpha expression in a porcine kidney cell line is independent of sphingoid base accumulation induced by ceramide synthase inhibition. AB - Previous studies have shown that fumonisin B1 (FB1) inhibits ceramide synthase, resulting in accumulation of free sphinganine and sphingosine. Tumor necrosis factor-alpha (TNFalpha) plays an important role in FB1 toxicity and the expression of TNFalpha mRNA in liver and kidney is increased following FB1 exposure in mice. The objective of the current study was to investigate whether these two events (sphingoid bases accumulation and TNFalpha induction) are dependent on each other. An increase in expression of TNFalpha mRNA was detected in LLC-PK1 cells as early as 4 h after FB1 treatment but decreased to the control levels after 8 h. A positive linear correlation was observed between the expression of TNFalpha mRNA and FB1 concentration. Increases of intracellular sphingoid bases were also detected after 4 h of FB1 treatment and progressively increased until 24 h. Exposure of the cells to sphinganine or sphingosine did not significantly alter the expression of TNFalpha. Inhibition of sphingoid base biosynthesis by ISP-1, a specific inhibitor of serine palmitoyltransferase, the first enzyme in de novo sphingolipid biosynthesis, efficiently blocked the accumulation of free sphingoid bases in response to FB1, but it did not prevent the induction of TNFalpha expression. Results indicate that FB1-induced increase in TNFalpha expression is independent of sphingoid base accumulation-induced by ceramide synthase inhibition in LLC-PK1 cells. PMID- 11437651 TI - Biodegradability of inhaled para-aramid respirable-sized fiber-shaped particulates: mechanistic in vivo and in vitro studies. AB - Biopersistence represents an important health-related issue in fiber toxicology. These studies were undertaken to elucidate the mechanism(s) through which inhaled p-aramid respirable-sized fiber-shaped particulates (RFP) are biodegraded in the lungs of exposed rats and hamsters. Previously, we and others have reported that, following deposition in the lung, long p-aramid RFPs are cleaved into shorter fibrous fragments. To investigate the mechanisms of RFP biodegradation, we have postulated that lung fluids coat/activate p-aramid RFP following deposition in the alveolar regions of the lung, thus predisposing the RFP to enzymatic attack and consequent shortening. This process enhances the rate of clearance of the inhaled RFP. To test this hypothesis, we have conducted both in vivo and in vitro cellular and noncellular investigations. First, p-aramid or cellulose RFP were instilled into the lungs of rats and the lungs were digested 24 h postexposure using two different digestion techniques: (1) a conventional ethanolic KOH method and (2) an enzymatic method that simulates the action of lung enzymes. Cellulose RFP were utilized as a control organic fiber-type that is known to be biopersistent. The results demonstrated that the enzymatic but not the KOH method resulted in transverse cleavage of the p-aramid RFP; the lengths of cellulose RFP recovered from rat lungs were not reduced by either method. Next, standardized preparations of p-aramid RFP or cellulose RFP were incubated with saline or lung fluids and then processed by one of two tissue digestion techniques. Mean lengths of p-aramid RFP incubated with saline and processed with KOH or the enzyme method were not found to be altered. Indeed, only the preparation of p-aramid RFP that had been incubated with bronchoalveolar lavage (BAL) fluids and processed with the enzyme solution resulted in cleavage of p-aramid RFP. Moreover, when the BAL fluids were autoclaved to denature proteins, the length dimensions of p-aramid RFP were intermediate between saline controls and RFP incubated with normal BAL fluids and processed via the enzymatic technique. In contrast to the in vitro noncellular studies with p-aramid RFP, the combination of BAL fluid incubation and enzyme digestion method had no measurable effect on shortening of cellulose RFP, indicating that the results with p-aramid were specific to that fiber-type. In a final set of in vitro cellular studies, cultures of rat lung epithelial cells, alveolar macrophages, or co-cultures of epithelial cells and macrophages were treated with p-aramid RFP for 1 h, 1 day, or 1 week to determine whether RFP shortening occurs directly in the phagocytic cells. The lengths of fibrils were measured using scanning electron microscopy techniques. The results demonstrated that (1) no shortening occurred in the epithelial cell cultures at any time point; however, (2) in the macrophage and cocultures, cleavage of p-aramid RFP was observed at 1 day and 1 week postexposure. Our data suggest that components of lung fluids coat and catalyze the p-aramid RFP as a prerequisite for enzymatic cleavage. This process could play a significant role in facilitating the transverse cleavage or shortening of inhaled p-aramid RFP in the lungs of exposed rats and hamsters. PMID- 11437652 TI - Productive infection of human primary cells and cell lines with porcine endogenous retroviruses. AB - Porcine endogenous retroviruses (PERVs) infect human cells in vitro and therefore represent a risk for xenotransplantation. However, first clinical transplantations of pig cells into humans or ex vivo perfusions did not result in transmission of PERVs. On the other hand, recent experiments with SCID mice demonstrated infections with PERV in vivo. In order to define and characterize human target cells, we studied numerous primary human cells and cell lines. Infection with PERVs was shown for human peripheral blood mononuclear cells, primary endothelial cells, and primary aortic smooth muscle cells as well as lymphocytic, monocytic, and epithelial cell lines. PMID- 11437653 TI - T cell apoptosis causes peripheral T cell depletion in mice transgenic for the HIV-1 vpr gene. AB - Vpr, an accessory protein of HIV, is known to affect viral replication as well as cell growth, differentiation, and apoptosis in vitro. To investigate its pathogenicity in vivo, we have produced mice transgenic for the HIV-1 vpr gene with the CD4 enhancer/promoter. Interestingly, apoptotic death of T lymphocytes was enhanced in those mice, causing marked reduction of T cells in lymphatic organs and peripheral blood. Involvement of Bcl-x, Bax, and Caspase-1, but not of the Fas-Fas ligand system, was suggested in the apoptotic processes. These observations suggest that Vpr is involved in the pathogenesis of T cell depletion in HIV-infected people. PMID- 11437654 TI - Impact of clinical reverse transcriptase sequences on the replication capacity of HIV-1 drug-resistant mutants. AB - We have shown that the HIV-1 laboratory strain NL4-3 that contains P236L [a reverse transcriptase mutation conferring resistance to the nonnucleoside reverse transcriptase inhibitor (NRTI) delavirdine] replicates more slowly than wild-type NL4-3. Other NNRTI-resistance mutations, such as K103N and Y181C, do not reduce the replication capacity of NL4-3 as much as P236L and develop more frequently in HIV-1 isolates from patients failing delavirdine. However, a minority of patients on delavirdine therapy still have isolates with P236L. We postulated that reverse transcriptase (RT) sequences from these patient isolates contain other mutations that compensate for the adverse effect of P236L. To test this hypothesis, we created 15 chimeric NL4-3 isolates that contained delavirdine-resistant RT sequences derived from eight patient isolates and characterized their replication kinetics. Nine of 10 patient-derived clones containing P236L replicated as slowly as NL4-3 with P236L. In contrast, three of five clones that did not have P236L (but had either K103N or Y181C) replicated significantly better than NL4-3 with P236L. Thus, the majority of patients who acquire P236L during delavirdine therapy do not have RT mutations that compensate for the replication defect conferred by P236L. We hypothesize that HIV-1 isolates with P236L may have a compensatory mutation outside RT. Alternatively, variants of HIV-1 with reduced replication fitness may be selected during antiretroviral therapy, suggesting that stochastic events rather than viral replication fitness may determine which drug-resistant mutants emerge early during antiretroviral failure. In some isolates, it appears that the background RT sequence can contribute significantly to the replication fitness of drug-resistant HIV-1 variants. PMID- 11437655 TI - Protection from immunodeficiency virus challenges in rhesus macaques by multicomponent DNA immunization. AB - Multicomponent DNA vaccines were used to elicit immune responses, which can impact viral challenge in three separate rhesus macaque models. Eight rhesus macaques were immunized with DNA vaccines for HIV env/rev and SIV gag/pol and were challenged intravenously with 10 animal infective doses (AID(50)) of cell free SHIV IIIB. Three of eight immunized rhesus macaques were protected, exhibiting no detectable virus. Animals protected from nonpathogenic SHIVIIIB challenge were rested for extended periods of time and were rechallenged first with pathogenic SIV(mac239) and subsequently with pathogenic SHIV89.6P viruses. Following the pathogenic challenges, all three vaccinated animals were negative for viral coculture and antigenemia and were negative by PCR. In contrast, the control animals exhibited antigenemia by 2 weeks postchallenge and exhibited greater than 10 logs of virus/10(6) cells in limiting dilution coculture. The control animals exhibited CD4 cell loss and developed SIV-related wasting with high viral burden and subsequently failed to thrive. Vaccinated animals remained virus-negative and were protected from the viral load, CD4 loss, disease, and death. We observed strong Th1-type cellular immune responses in the protected macaques throughout the study, suggesting their important roles in protection. These studies support the finding that multicomponent DNA vaccines can directly impact viral replication and disease in a highly pathogenic challenge system, thus potentially broadening our strategies against HIV. PMID- 11437656 TI - The poliovirus receptor CD155 mediates cell-to-matrix contacts by specifically binding to vitronectin. AB - The human receptor for poliovirus (CD155) is an immunoglobulin-like molecule with unknown normal function(s). Here we provide evidence that CD155 binds specifically to vitronectin with a dissociation constant (K(d)) of 72 nM as determined by surface plasmon resonance. Based on sequence homology to the CD155 gene, three poliovirus receptor-related genes (PRR1, PRR2, and PRR3) were cloned recently. PRR proteins were reported by others to mediate homophilic cell adhesion. Neither PRR1 nor PRR2 binds poliovirus and it is assumed that their physiological functions differ from that of CD155. Indeed, mPRR2 was found to bind to vitronectin only weakly, while its self-adhesion activity is characterized by a K(d) of 310 nM. Moreover, there is no evidence for CD155 self adhesion. Both CD155 and vitronectin colocalize to follicular dendritic cells and B cells inside the germinal centers of secondary lymphoid tissue (tonsils)-an observation suggesting that the CD155/vitronectin interaction is required for the establishment of a proper immune response in this particular context. PMID- 11437657 TI - White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp. AB - White spot syndrome virus (WSSV) is a large DNA virus infecting shrimp and other crustaceans. The virus particles contain at least five major virion proteins, of which three (VP26, VP24, and VP15) are present in the rod-shaped nucleocapsid and two (VP28 and VP19) reside in the envelope. The mode of entry and systemic infection of WSSV in the black tiger shrimp, Penaeus monodon, and the role of these proteins in these processes are not known. A specific polyclonal antibody was generated against the major envelope protein VP28 using a baculovirus expression vector system. The VP28 antiserum was able to neutralize WSSV infection of P. monodon in a concentration-dependent manner upon intramuscular injection. This result suggests that VP28 is located on the surface of the virus particle and is likely to play a key role in the initial steps of the systemic WSSV infection in shrimp. PMID- 11437658 TI - Identification of dna components required for induction of cotton leaf curl disease. AB - Cotton leaf curl disease (CLCuD) is a major constraint to cotton production in Pakistan. Infectious clones of the monopartite begomovirus cotton leaf curl virus (CLCuV), associated with diseased cotton, are unable to induce typical symptoms in host plants. We have identified and isolated a single-stranded DNA molecule approximately 1350 nucleotides in length which, when coinoculated with the begomovirus to cotton, induces symptoms typical of CLCuD, including vein swelling, vein darkening, leaf curling, and enations. This molecule (termed DNA beta) requires the begomovirus for replication and encapsidation. The CLCuV/DNA 1/DNA beta complex, together with a similar complex previously identified in Ageratum conyzoides, represent members of an entirely new type of infectious, disease-causing agents. The implications of this finding to our understanding of the evolution of new disease-causing agents are discussed. PMID- 11437659 TI - The transcription/replication activity of the polymerase of influenza A viruses is not correlated with the level of proteolysis induced by the PA subunit. AB - The PA subunit of the influenza virus polymerase has been shown to induce degradation of coexpressed proteins, but its role in the replication activity of the polymerase is not fully understood. Here, PA proteins derived from several influenza A viruses were examined at 37 and 33 degrees C for both the level of proteolysis they induced and the efficiency with which they ensured transcription/replication of a viral-like RNA within a polymerase complex reconstituted in vivo from cloned cDNAs. Two mutants of A/Victoria/3/75 PA showed a decreased ability to induce proteolysis as compared to the wild-type PA, but still appeared to be as active as the wild-type protein with respect to the polymerase activity. Furthermore, we observed that the ability of PR8-PA to induce proteolysis was severely impaired at 33 degrees C as compared to 37 degrees C, while the efficiency with which the PR8-derived polymerase complex ensured transcription/replication of the viral-like RNA was similar at both temperatures. Taken together, our observations suggest that the transcription/replication activity of the polymerase of influenza A viruses is not correlated with the level of proteolysis induced by the PA subunit. PMID- 11437660 TI - Deletion and substitution analysis defines regions and residues within the phosphoprotein of bovine respiratory syncytial virus that affect transcription, RNA replication, and interaction with the nucleoprotein. AB - The phosphoprotein (P) of bovine respiratory syncytial virus (BRSV) is a multifunctional protein that plays a central role in transcription and replication of the viral genomic RNA. To investigate the domains and specific residues involved in different activities of the P protein, we generated a total of 22 deletion and 17 point mutants of the P protein. These mutants were characterized using an intracellular BRSV-CAT minigenome replication system for the ability to (1) direct minigenome transcription, (2) direct minigenome replication, and (3) form complexes with nucleocapsid protein (N) and large polymerase protein (L). These studies revealed that all the regions of P protein except amino acids 41-80 are essential for minigenome transcription and replication. Interestingly, amino acids 41-60 appeared to contain sequences that negatively regulate transcription and replication. Analysis of the N- or C terminal ends indicated that deletion of up to 3 amino acids from the N- or C terminus completely ablated the replication, while leaving substantial residual transcription. Single amino acid substitutions within the N-terminal 4 or C terminal 13 amino acids showed that substitution at position 2, 4, 234, 236, 238, 240, or 241 was highly inhibitory to both transcription and replication, whereas substitution at position 3 was highly inhibitory to replication while leaving substantial residual transcription. Substitution of serine residues at the C terminus indicated that loss of phosphorylation sites did not appear to have any effect on transcription and replication. Coimmunoprecipitation of P-N and P-L complexes with P-specific antiserum revealed that substitution mutations at the N or C-terminus did not affect binding to N and L proteins, except that substitution mutation at C-terminus position 234, 236, 238, 240, or 241 affected binding to N protein by 10-fold. PMID- 11437661 TI - A plasmid of phytoplasma encodes a unique replication protein having both plasmid and virus-like domains: clue to viral ancestry or result of virus/plasmid recombination? AB - The genomes of most prokaryotic and eukaryotic single-stranded (ss) DNA viruses, and some prokaryotic plasmids such as pLS1, commonly replicate via a rolling circle replication (RCR) strategy, and thus the viruses are hypothesized to have evolved from the plasmids, although evidence for this view is sparse. We have sequenced a circular plasmid of 3933 nt, pOYW, obtained from onion yellows phytoplasma (OY-W), a cell-wall-less, unculturable prokaryote that inhabits the cytoplasm of both plant and insect cells. pOYW contains five open reading frames (ORFs) on the same strand and apparently replicates by an RCR mechanism. Its rep gene (ORF5) encodes a unique protein, pOYW-Rep, with an unprecedented structure. The N-terminal region of pOYW-Rep has similarities to the RCR initiator protein (Rep) of pLS1 family plasmids but, unlike the Rep of other plasmids, its C terminal region was unexpectedly similar to the helicase domain of the replication-associated proteins (Rap) of eukaryotic viruses, especially circoviruses (ssDNA viruses of vertebrates). The pOYW-Rep was specifically detected in OY-W-infected plant phloem cells, suggesting that it is a functional protein. We suggest that an ancestral phytoplasma plasmid pOYW may have acquired a helicase domain from host phytoplasmal DNA, entered the surrounding eukaryotic cytoplasm, and subsequently evolved into an ancestral eukaryotic ssDNA virus. Alternatively, a pOYW ancestor could have obtained the helicase domain by recombination with a virus: this would be the first example of recombination between plasmids and viruses. PMID- 11437662 TI - Self-priming of retroviral minus-strand strong-stop DNAs. AB - After minus-strand strong-stop DNA (-sssDNA) is synthesized, the RNA template is degraded by the RNase H activity of the reverse transcriptase (RT), generating a single-stranded DNA. The 3' end of -sssDNA from HIV-1 can form a hairpin; this hairpin will self-prime in vitro. We previously used a model substrate, -R ssDNA, which corresponds to the 3' end of the -sssDNA of HIV-1, to show that the self priming of this model substrate could be prevented by annealing a 17-nt-long DNA oligonucleotide to the 3' end of -R ssDNA in the presence of HIV-1 nucleocapsid (NC) protein. Similar model substrates were prepared for HIV-2 and HTLV-1; the R regions of these two viruses are longer and form more complex structures than the R region of the HIV-1 genome. However, the size of the R region and the complexity of the secondary structures they can form do not affect self-priming or its prevention. The efficiency of the self-priming is related to the relative stabilities of the conformations of -R ssDNA that can and cannot induce self priming. For the three viruses (HIV-1, HIV-2, and HTLV-1), the size of the DNA oligonucleotide needed to block self-priming in the presence of NC is similar to the expected size of the piece of RNA left after degradation of the RNA template during reverse transcription. We also found that when the 3' end of -R ssDNA is annealed to a complementary DNA oligonucleotide, it is a good substrate for efficient nonspecific strand transfer to other single-stranded DNA molecules. PMID- 11437663 TI - The gene encoding the major viral structural protein stimulates recombination in polyomavirus DNA. AB - RmI is a chimeric DNA molecule consisting of a polyoma genome in which a partly duplicated VP1-coding region brackets an insert of murine DNA (Ins); when transfected into mouse cells, RmI recombines intramolecularly to yield infectious, unit-length, polyoma DNA. We report here that RmI encodes a polypeptide of 337 amino acids (designated VmP1) which includes the N-terminal 328 amino acids of VP1 and 9 amino acids specified by Ins. Mutating the VmP1 coding sequence strongly reduces the ability of RmI to yield polyoma DNA. In contrast, mutating the portion of the VP1-coding sequence which is not part of the VmP1-coding sequence has little or no impact on the ability of RmI to yield polyoma DNA, even though it renders such DNA noninfectious. Thus, release of polyoma DNA from RmI involves a function of VP1 distinct from that ensuring virus assembly and propagation; since VP1 can arise only after recombination has occurred, VmP1, but not VP1, could carry such a function. We suggest that VmP1 acts in concert with VP2, which we have already reported to stimulate recombination in RmI. PMID- 11437664 TI - Structure of the fiber head of Ad3, a non-CAR-binding serotype of adenovirus. AB - Adenoviruses of serotype Ad3 (subgenus B) use a still-unknown host cell receptor for viral attachment, whereas viruses from all other known subgenera use the coxsackie and adenovirus receptor (CAR). The receptor binding domain (head) of the Ad3 fiber protein has been expressed in Escherichia coli inclusion bodies. After denaturation and renaturation using a rapid dilution method, crystals of trimeric head were obtained. The 1.6 A resolution X-ray structure shows a strict conservation of the beta-sheet scaffold of the protein very similar to the head structures of the CAR-binding serotypes Ad2, Ad5, and Ad12. The conformation of the loops is different, with the exception of the AB loop, which forms the center of the interface in the Ad12-CAR complex structure. The structure explains why a mutation in Ad5 of one residue in the AB loop to glutamic acid, as in Ad3, abrogates binding to CAR. It is possible that the Ad3 receptor binding site is nevertheless situated similar to the CAR binding site, although it cannot be excluded that other regions of the relatively hydrophobic head surface may be used. PMID- 11437665 TI - Intron conservation in the DNA polymerase gene encoded by Chlorella viruses. AB - Previously we reported that 19 of 42 viruses that infect Chlorella strain NC64A (NC64A viruses) contain a short, nuclear-located, spliceosomal-processed intron in a pyrimidine dimer-specific glycosylase/apyrimidine lyase (pdg) gene. Surprisingly, the nucleotide sequence of the intron region is more conserved than the exon regions of the gene (L. Sun et al., 2000, J. Mol. Evol. 50, 82-92). For comparative purposes, we determined the nucleotide sequence of a similar intron type and its flanking coding regions in the DNA polymerase (dnapol) gene from the same 42 NC64A viruses and also 5 viruses that infect Chlorella strain Pbi. Thirty eight of the 42 NC64A viruses contained a 101-nucleotide intron and the remaining 4 had an 86-nucleotide intron located in the same position in dnapol. The 4 viruses with the smaller intron in dnapol also have a smaller intron in their pdg gene. There was no intron in the dnapol gene of the 5 Pbi viruses. Phylogenetic analyses indicate that the dnapol genes containing the 86-nucleotide intron represent the ancestral condition among the NC64A viruses. The intron in the dnapol gene is phase 0 (keeps codons intact), which differs from the phase 1 intron in the pdg gene. The intron in the dnapol gene, unlike the pdg intron, was conserved (83 to 100% identical) to about the same extent as the coding regions of the gene (78 to 100% identical). PMID- 11437666 TI - Mutations in the GDD motif of rubella virus putative RNA-dependent RNA polymerase affect virus replication. AB - Rubella virus (RV) nonstructural proteins are translated as a p200 polyprotein that undergoes proteolytic cleavage into p150 and p90. From conserved amino acid sequence motifs in polypeptides, p90 has been proposed to be the RV RNA-dependent RNA polymerase (RdRp). To test whether the conserved GDD motif is involved in RdRp catalytic activity, three different alanine substitutions were introduced into it. Substitution of glycine by alanine (G1966A) resulted in impaired virus infectivity. Alteration of either aspartate residue completely abolished virus replication. A fully infectious variant was isolated from the G1966A mutant. Sequencing analysis showed that the alanine residue substituted in G1966A mutant had reverted to glycine in this variant. Complementation experiments were carried out to rescue the replication-defective RNA carrying G1966A, D1967A, or D1968A mutations. The defective RNA with G1966A mutation in p90 replicated efficiently when the helper genome that supplied a wild-type p90 was provided in trans. However, the replication-defective RNA with D1967A or D1968A was not rescued by supplementation of p90 in trans. Our studies support the idea that the GDD motif is critical for RV replication and p90 function as RV RdRp. PMID- 11437667 TI - Molecular and genetic evidence for a virus-encoded glycosyltransferase involved in protein glycosylation. AB - The major capsid protein, Vp54, of chlorella virus PBCV-1 is a glycoprotein that contains either one glycan of approximately 30 sugar residues or two similar glycans of approximately 15 residues. Previous analysis of PBCV-1 antigenic mutants that contained altered Vp54 glycans led to the conclusion that unlike other glycoprotein-containing viruses, most, if not all, of the enzymes involved in the synthesis of the Vp54 glycan are probably encoded by PBCV-1 (I.-N. Wang et al., 1993, Proc. Natl. Acad. Sci. USA 90, 3840-3844). In this report we used molecular and genetic approaches to begin to identify these virus genes. Comparing the deduced amino acid sequences of the putative 375 PBCV-1 protein encoding genes to databases identified seven potential glycosyltransferases. One gene, designated a64r, encodes a 638-amino-acid protein that has four motifs conserved in "Fringe type" glycosyltransferases. Analysis of 13 PBCV-1 antigenic mutants revealed mutations in a64r that correlated with a specific antigenic variation. Dual-infection experiments with different antigenic mutants indicated that viruses that contained wild-type a64r could complement and recombine with viruses that contained mutant a64r to form wild-type virus. Therefore, we conclude that a64r encodes a glycosyltransferase involved in synthesizing the Vp54 glycan. This is the first report of a virus-encoded glycosyltransferase involved in protein glycosylation. PMID- 11437668 TI - The NS2 protein generated by the parvovirus minute virus of mice is degraded by the proteasome in a manner independent of ubiquitin chain elongation or activation. AB - The NS2 protein generated by the parvovirus minute virus of mice is very labile, having a half-life during infection of approximately 90 min. The degradation of NS2 is blocked by proteasome inhibitors but is likely ubiquitin independent: NS2 does not form detectable higher molecular weight ubiquitin-containing conjugates, and NS2 degradation requires neither ubiquitin chain elongation nor intracellular ubiquitin activation. We have also identified a region in the carboxyl half of NS2 that is required for its proteasome-mediated degradation. PMID- 11437669 TI - CD4(+) and CD8(+) T cell death during human immunodeficiency virus infection in vitro. AB - We have evaluated the death of CD4(+) and CD8(+) T cells during in vitro human immunodeficiency virus (HIV) infection of peripheral blood mononuclear cells (PBMC) and tonsilar tissue. Acute infections with several X4 and R5 HIV isolates induced a decrease in cell viability that was higher in infections with X4 viruses and correlated with an increased rate of CD4(+) T-cell death. In CD4(+) T cells, the primary X4 isolate AOM induced higher levels of death than the laboratory X4 isolates IIIB and NL4-3 or the R5 isolates BaL and MDM. An effect on CD8(+) T-cell viability was exclusively observed in infections by X4 viruses, including the NL4-3 strain, in both PBMC and tonsilar tissue. This effect was dependent on the env gene of the infecting isolate and required productive HIV replication in CD4(+) but not in CD8(+) T cells. Our results suggest that X4 and R5 HIV isolates depleted CD4(+) T cells to a different extent and that CD8(+) T cell viability may also be affected by mechanisms other than those acting in CD4(+) T cells. PMID- 11437670 TI - Use of chimeric nectin-1(HveC)-related receptors to demonstrate that ability to bind alphaherpesvirus gD is not necessarily sufficient for viral entry. AB - Human nectin-1 (HveC, Prr1), a member of the immunoglobulin superfamily and a receptor for the entry of herpes simplex viruses 1 and 2 (HSV-1, HSV-2), pseudorabies virus (PRV), and bovine herpesvirus 1 (BHV-1), binds to viral gD. For HSV-1, HSV-2, and PRV, the gD-binding region of nectin-1 has been localized to the N-terminal V-like domain. To determine whether the two C-like domains of nectin-1 influenced gD binding and entry activity, genes encoding chimeric proteins were constructed. Portions of nectin-1 were replaced with homologous regions from nectin-2 (HveB, Prr2), a related protein with ability to mediate the entry of PRV, HSV-2, and Rid mutants of HSV-1, but not HSV-1 or BHV-1. Also, one or more domains of nectin-1 were fused to the two membrane-proximal Ig domains of CD4, a protein with no herpesvirus entry or gD-binding activity. The chimeric proteins were expressed in Chinese hamster ovary cells, which normally lack alphaherpesvirus entry receptors, and detected on the cell surface by one or more anti-nectin-1 monoclonal antibodies. One chimeric protein (nectin-1 amino acids 1 124 fused to CD4) failed to bind to soluble forms of HSV-1, HSV-2, PRV, and BHV-1 gD and, as expected, also failed to mediate entry of the viruses from which these gDs were derived. The other chimeric receptors bound all forms of gD. Some mediated the entry of all the viruses tested but others mediated entry of some but not all the viruses. We conclude that binding of gD to the nectin-1 V domain is not sufficient for entry activity, that there are structural requirements for entry activity independent of gD binding, and that these requirements are different for the several alphaherpesviruses that can use nectin-1 as a receptor. PMID- 11437671 TI - Potential adverse effects of cyclooxygenase-2 inhibition: evidence from animal models of inflammation. AB - Cyclooxygenase (COX; prostaglandin H synthase, prostaglandin endoperoxidase) is the key enzyme in the synthesis of the prostaglandin and thromboxane families of eicosanoid mediators, and is the target for the nonsteroidal anti-inflammatory drugs (NSAIDs). The identification of an inducible COX isoform, COX-2, and the demonstration of its specific expression at sites of inflammation suggested that it may provide a useful therapeutic target for novel anti-inflammatory drugs. Inhibition of an enzyme that is not expressed in most healthy tissues would potentially avoid most of the adverse effects associated with NSAIDs, which target a constitutively expressed isoform, COX-1. The development of novel 'super aspirins' with high selectivity towards the inhibition of COX-2 showed that this hypothesis was well-founded and that high levels of these drugs could be tolerated without these serious adverse effects. The first two of these new generation NSAIDs, celecoxib and rofecoxib, are now in clinical use. More recently, however, concern has been expressed that COX-2 inhibition may in fact have a number of potential, previously hidden, pitfalls. These have arisen from the demonstration that COX-2 induction is not exclusively associated with the onset of an inflammatory reaction, with expression limited to inflammatory sites. In fact, COX-2 is expressed more chronically, and is also seen during the resolution of inflammation and in areas of wound-healing. The application of COX 2-selective inhibitors during these periods has been shown to be deleterious in that resolution of inflammation is delayed, gastric ulcer healing is delayed and, in some patients, ulcers have been shown to progress further to perforation. The suggestion has now been made that, in these situations, COX-2 may help resolve the pathology, perhaps by generating alternative series of prostaglandins such as the cyclopentenone prostaglandins. The finding that these prostaglandins can affect proteins by direct chemical modifications as well as having their own receptor families has rekindled debate on the deleterious and beneficial effects of prostanoids, and the implications of inhibiting the production of these mediators, in the body. Therefore, in this review we discuss the role of COX-2 in inflammation and the potential adverse effects of its inhibition. PMID- 11437672 TI - Long-acting beta-agonist treatment in patients with persistent asthma already receiving inhaled corticosteroids. AB - International guidelines recommend that long-acting beta-agonists should be considered in patients who are symptomatic despite moderate doses of inhaled corticosteroids. When combined with inhaled corticosteroids they improve asthma symptoms and lung function and reduce exacerbations. The evidence suggests that they are well tolerated. However, they are less effective than inhaled corticosteroids as monotherapy and should not be used alone, although the addition of a long-acting beta-agonist may permit a small reduction in the corticosteroid dose. Both salmeterol and formoterol appear equally effective in improving asthma control. Formoterol, however, has a rapid onset of action and is now being promoted for the relief of acute asthma symptoms. Both drugs provide prolonged protection against exercise-induced bronchospasm. However, this effect rapidly diminishes with continuous therapy and if this is the main aim of treatment, intermittent use may be preferable. When compared with alternative treatments, inhaled long-acting beta-agonists are more effective in controlling asthma symptoms than either theophylline or antileukotriene agents. Bambuterol, an oral prodrug of terbutaline, appears to be as effective as the inhaled long acting beta-agonists and has the advantage of once daily oral administration. However, the inhaled long-acting beta-agonists are less likely to have systemic adverse effects. There are theoretical concerns that regular beta-agonist treatment may lead to tolerance and a failure to respond to emergency asthma treatment. While there is no doubt that tolerance occurs, there is currently little evidence that this is a clinical problem. Insights into pharmacological as well as therapeutic interactions between inhaled corticosteroids and beta agonists are providing justification for their use in combination. Guidelines for the management of patients with chronic persistent asthma are likely to require modification to reflect these developments. PMID- 11437673 TI - Potential new therapeutic options in Behcet's syndrome. AB - Behcet's syndrome is a multisystem disorder that causes orogenital ulceration, skin lesions and intraocular inflammation with uveitis and retinal vasculitis. A proportion of affected individuals also develop vascular and central nervous system manifestations, with significant morbidity and mortality. Although the aetiopathogenesis of Behcet's syndrome is poorly understood, the condition is considered to be driven, at least in part, by autoimmune mechanisms. Conventional therapy relies on available anti-inflammatory and immunomodulatory agents, and, in view of the paucity of controlled clinical trials, it is to a large extent empirical. Oral ulcers can often be treated by topical application of corticosteroids. In addition to corticosteroids, agents used to treat ocular inflammation and significant systemic manifestations include colchicine, thalidomide, azathioprine, mycophenolate mofetil, cyclosporin, tacrolimus, cyclophosphamide and chlorambucil. The response to these agents is variable and there is a distinct need for more effective rational treatment. Over the last decade, a number of open studies have produced promising results using recombinant interferon-alpha preparations. Evaluating, in a methodical manner, the other new biological agents that are becoming available for the treatment of inflammatory diseases offers great promise, not only for effective management but also for providing insights into aetiopathogenesis. PMID- 11437674 TI - Mycophenolate mofetil: suggested guidelines for use in kidney transplantation. AB - Mycophenolate mofetil (MMF) is an immunosuppressive drug designed to inhibit inosine monophosphate dehydrogenase (IMPDH). IMPDH is a key enzyme in the de novo purine synthesis of lymphocytes. It is crucially important for proliferative responses of human T and B lymphocytes. The inhibition of IMPDH thus leads to selective lymphocyte suppression. After successful use in various in vitro and animal models, MMF was brought to clinical trial in patients undergoing transplantation. The drug is rapidly and completely absorbed following oral administration. Pilot studies of administration with cyclosporin and corticosteroids suggested a significant reduction in the incidence of organ rejection at dosages of 1 to 3 g/day. As a result of these studies, 3 pivotal randomised double-blind multicentre trials, involving nearly 1500 patients, were designed to investigate the effects of addition of MMF to different standard immunosuppressive protocols on the prevention of acute renal allograft rejection. After 6 months, the rates of biopsy-proven rejection were significantly reduced in patients receiving MMF. In combination with cyclosporin and corticosteroids, the adverse effect profile resembled that of azathioprine. Most adverse effects were associated with the gastrointestinal tract, the blood system and opportunistic infections. MMF offers improved immunosuppressive therapy following renal and probably other solid organ transplantation. MMF has been licensed since 1995 for the prevention of acute renal allograft rejection in most countries. It has been used in different combinations of immunosuppressive drugs and in various dosages and regimens. PMID- 11437675 TI - Novel therapeutics for chemotherapy-resistant acute myeloid leukaemia. AB - Patients with chemotherapy-resistant acute myeloid leukaemia are rarely cured by non-allogeneic transplant therapies. Multiple new investigational agents have become available for treatment of these patients and there are few tools to permit rational drug and clinical trial selection. In this review, we describe the chemical and biological properties of some of these agents and some of their initial clinical activity to date. The selected agents react with either cell surface molecules or signal pathway intermediates and include antibody and antibody conjugates to CD33 and CD45, a fusion protein directed to the granulocyte-macrophage colony-stimulating factor receptor, an anti-sense oligonucleotide to Bcl2, a farnesyl transferase inhibitor, and a protein kinase C agonist/inhibitor. The challenge for the next decade will be how to select patients for particular molecularly targeted therapeutics and how to combine these agents. PMID- 11437676 TI - Role of B cells in the pathogenesis of rheumatoid arthritis: potential implications for treatment. AB - In patients with rheumatoid arthritis, chronic inflammation in affected joints may lead to the development of tertiary lymphoid tissue. A micro-environment is generated in the synovial membrane which supports the activation and differentiation of B cells into plasma cells. Through a process of affinity maturation, plasma cells may be generated locally which secrete antibodies of high affinity. Rheumatoid arthritis is characterised by autoantibodies specific for self immunoglobulin. These rheumatoid factors form large antigen/antibody complexes which may enhance the process of joint destruction. The poor prognosis of rheumatoid factor-positive patients is indicitive of the critical role of immunoglobulin complexes in the continuous stimulation of the immune system and thus of the inflammatory processes. In general, treatment of patients with rheumatoid arthritis aims at suppressing inflammation. The currently most successful reagents are those which interfere with the network of cytokines, such as tumour necrosis factor or interleukin-1 receptor antagonists. Only recently have immunosuppressive therapies targeted directly at the B cell response been developed. These first studies suggest that therapies which directly affect the humoral immune response are of great therapeutic potential in the treatment of patients with rheumatoid arthritis. PMID- 11437677 TI - The potential role of tocopherol in asthma and allergies: modification of the leukotriene pathway. AB - Metabolism of arachidonic acid via the 5-lipoxygenase (5-LO) pathway leads to the formation of hydroperoxyeicosatetraenoic acids (HPETEs) and leukotriene (LT) A4. This unstable allylic epoxide can be further converted by secondary enzymes into LTB(4) and cysteinyl LTs. LTs represent a family of potent biologically active compounds synthesised by specific cell types and by transcellular biosynthetic mechanisms. Cysteinyl LTs are involved in the pathogenesis of asthma, and recent data indicate that individuals with asthma may have enhanced basal excretion of urinary LTE4 compared with normal individuals. Tocopherol (vitamin E) and tocopherol acetate strongly inhibit potato 5-LO in an irreversible and noncompetitive way, and, by affecting the redox state of cells possessing 5-LO, they may influence the production of biologically active LTs. It has been reported that normal plasma levels of tocopherol may enhance the lipoxygenation of arachidonic acid, whereas higher tocopherol levels exert a suppressive effect that is consistent with its role as a hydroperoxide scavenger. Receptor-mediated activation of neutrophils in individuals with asthma results in the synthesis of LTs. This activation is inhibited by tocopherol in a concentration-dependent manner. Additional controlled studies are needed to assess the effect of tocopherol on leukotriene production in asthmatic individuals. The results of these studies may be useful in developing new therapeutic approaches in asthmatic/allergic patients. PMID- 11437678 TI - The prospect of treating rheumatoid arthritis with recombinant human interleukin 1 receptor antagonist. AB - The prospect of introducing a potent novel therapy that specifically targets a pivotal mediator of disease offers new hope to patients with rheumatoid arthritis (RA). There is convincing evidence that interleukin-1 (IL-1) plays a critical role in the pathogenesis of RA. In RA, the inadequate production of IL-1 receptor antagonist (IL-1Ra), the natural inhibitor of IL-1, results in increased IL-1 mediated pathophysiological activity. Unregulated IL-1-mediated effects produce enhanced tissue inflammation and progressive degradation of cartilage and bone matrix. Recombinant IL-1Ra treatment in experimental models of arthritis results in profound suppression of synovial inflammation and joint damage. Recombinant human IL-1Ra (rhu-IL-1Ra) has been evaluated in RA in several randomised clinical trials and was shown to significantly reduce both the clinical manifestations of arthritis and the rate of progressive joint damage. Firstly, in a 6-month placebo controlled study, 43% of the patients who received the highest dose of rhu-IL-1Ra (150 mg/day) demonstrated a 20% improvement (American College of Rheumatology clinical criteria) compared to 27% of the patients who received placebo. In addition, the patients who received rhu-IL-1Ra demonstrated 46% less joint damage (Larsen scores). Secondly, in a 6-month extension of the placebo-controlled study, the treated patients maintained their clinical improvement and there was further significant reduction in the rate of progressive joint damage. The patients who had originally received placebo demonstrated both clinical and radiological responses that were similar to those observed in the treated patients during the initial phase of the study. Finally, in a combination study with methotrexate (MTX), 42% of patients who received MTX and the optimal dose of rhu-IL-1Ra (1.0 mg/kg/day) demonstrated an ACR 20% clinical response, compared to 23% of those receiving MTX and placebo. rhu-IL-1Ra was well tolerated in all studies and adverse events were uncommon. The most frequently reported adverse event causing withdrawal of treatment was an injection site reaction, occurring in approximately 5% of patients. In conclusion, targeted IL-1 inhibition significantly reduced both the clinical manifestations and the rate of progressive joint damage in patients with RA. These observations suggest that rhu IL-1Ra has a potential role as a novel therapeutic modality in the future management of RA. PMID- 11437679 TI - Schonlein-henoch purpura in children and adults: diagnosis, pathophysiology and management. AB - Schonlein-Henoch syndrome is a form of systemic small-vessel vasculitis, characterised by vascular and/or mesangial immunoglobulin A1 deposits. The main clinical manifestations are vascular purpura, predominating on the lower limbs, and articular, gastrointestinal and renal symptoms. Pulmonary, cardiac, genital and neurological symptoms have also be observed. The syndrome usually affects children, whereas it is rare in adults. The frequency of renal involvement varies between individual studies (from 20 to 100%). Renal manifestations are usually mild and transient, although chronic nephropathies may occur. Overall, an estimated 2% of children with Schonlein-Henoch purpura progress to renal failure and up to 20% of children with nephritis treated in specialised centres require haemodialysis. The renal prognosis appears to be worse in adults. Aetiological investigations are required, as a triggering factor is found in approximately half the patients (e.g. viral, bacterial and parasitic infections, drugs, toxins, systemic diseases and cancer). Dapsone has beneficial effects on cutaneous, gastrointestinal and articular manifestations in adults, especially those with chronic forms. Corticosteroids may be useful for refractory abdominal pain. Methylprednisolone pulse therapy, immunosuppressive drugs (e.g. cyclophosphamide and azathioprine), plasma exchange and polyclonal immunoglobulin therapy are beneficial in very rare life-threatening forms of the disease and in rare instances where renal function is compromised. PMID- 11437680 TI - Targeting tumour necrosis factor in the treatment of rheumatoid arthritis. AB - Inflammation in rheumatoid arthritis (RA) is associated with an imbalance between pro- and anti-inflammatory factors, which leads to a persistent chronic inflammatory state in the joint. Molecular studies of the physiology of the inflammatory response have identified a hierarchy of cytokine activities. The identification of this hierarchy has provided new potential therapeutic targets for the treatment of RA. At present the majority of new therapeutic agents have been developed to neutralise the activity of tumour necrosis factor-alpha (TNF alpha), a cytokine at the top of the inflammatory cascade. These agents consist of recombinant proteins that bind and neutralise TNF alpha, and they are effective in the treatment of inflammation in RA. In this review we discuss the rationale behind targeting TNF alpha, the various recombinant proteins that have been used, their clinical effectiveness, the possible adverse effects of these agents and the development of new chemical inhibitors of TNF alpha synthesis. PMID- 11437681 TI - Radioimmunotherapy: potential as a therapeutic strategy in non-Hodgkin's lymphoma. AB - Lymphomas are the fifth most common malignancy in the United States and are increasing in incidence. Despite being among the most responsive malignancies to radiation and chemotherapy, the majority of patients relapse or have progressive disease. Monoclonal antibodies (MAbs) directed at cell-specific surface antigens have been useful in the diagnosis of lymphomas and, more recently, the therapeutic mouse-human chimeric MAb rituximab has demonstrated effectiveness in B cell lymphomas. Conjugating MAbs to radionuclides is a strategy for improving the efficacy of MAb lymphoma therapy by delivering radiation in close proximity to the tumour (radioimmunotherapy or RIT). In addition, the low dose rate of the delivered radiation may exert a greater antitumour activity than an equivalent dose of conventional external beam radiation. The antigenic targets for MAb therapy have included CD20, CD22, HLA-DR, and B cell idiotype. Radionuclides that have been used include iodine-131, yttrium-90, and copper-67; there are relative merits and disadvantages to each source of radiation. Clinical studies to date have focused on relapsed and refractory patients with both indolent and aggressive lymphomas, although more recent studies have included previously untreated patients with indolent lymphoma. Radioimmunoconjugate has been delivered as either single or multiple doses. Response rates have varied widely, dependent on the patient population and the response criteria. Of note, complete responses can be achieved in this typically refractory patient group. Toxicities have generally consisted of mild infusion-related nausea, fever, chills, and asthenia. Neutropenia and thrombocytopenia are the dose-limiting toxicities and have prompted the incorporation of autologous stem cell support as a means of achieving dose escalation. To date, RIT has been delivered to highly selected patients in relatively few centres with requisite equipment and specialised personnel. In addition to these requirements, cost is likely to be a barrier to widespread use. The combination of RIT with chemotherapy at conventional or high dose, or with biological agents is a fertile area for investigation. The potential of RIT in the treatment for lymphomas will be defined only by well designed comparative prospective clinical studies. PMID- 11437682 TI - Thalidomide in cancer: potential uses and limitations. AB - In addition to immunomodulatory and cytokine-modulatory properties, thalidomide has antiangiogenic activity. It has been investigated in a number of cancers including multiple myeloma, myelodysplastic syndromes, gliomas, Kaposi's sarcoma, renal cell carcinoma, advanced breast cancer, and colon cancer. Its role has been best explored in myeloma, where, at daily doses of 100 to 800 mg, it is remarkably active, causing clinically meaningful responses in one-third of extensively pretreated patients and in over half of patients treated early in the course of the disease. It also acts synergistically with corticosteroids and chemotherapy in myeloma. Thalidomide produces improvement of cytopenias characteristic of myelodysplastic syndrome, resulting in the reduction or elimination of transfusion dependence in some patients. Responses have also been seen in one-third of patients with Kaposi's sarcoma, in a small proportion of patients with renal cell carcinoma and high grade glioma and, in combination with irinotecan, in some patients with colon cancer. Thalidomide is being investigated currently in a number of clinical trials for cancer. Drowsiness, constipation and fatigue are common adverse effects seen in 75% of patients. Symptoms of peripheral neuropathy and skin rash are seen in 30%. A minority of patients experience bradycardia and thrombotic phenomena. Despite the high frequency of adverse effects, those severe enough to necessitate cessation of therapy are seen in only 10 to 15% of patients. A therapeutic trial of thalidomide should be considered in all patients with myeloma who are unresponsive to or relapse after standard therapy. In other malignant diseases, the most appropriate way to use the drug is in the setting of well designed clinical trials. In the absence of access to such studies, thalidomide could be considered singly or in combination with standard therapy in patients with no meaningful therapeutic options. PMID- 11437683 TI - Myasthenia gravis: options and timing of immunomodulatory treatment. AB - The autoimmune pathogenesis of myasthenia gravis is relatively well understood. The current options for treatment of this disease are acute and long term immunotherapies, acetylcholinesterase inhibitors and thymectomy. Many factors influence the timing of initiation of immunomodulatory therapy in myasthenia gravis and both disease factors, such as stage and severity, and patient factors, such as age, pregnancy and intercurrent illness, must be considered. Decisions regarding the choice of therapy can be difficult because of the limited number of randomised controlled trials that have been performed in myasthenic patients. In general, acetylcholinesterase inhibitors alone are used only in mild ocular disease, and in the majority of other patients immunomodulatory therapy is begun early. Corticosteroids are the most commonly used initial therapy, followed by azathioprine. In refractory cases, the available options include immunosuppressants such as cyclosporin, mycophenolate mofetil and cyclophosphamide. Plasmapheresis and intravenous immunoglobulin are important in the treatment of acute exacerbations and myasthenic crisis and in the perioperative setting. Despite many years of experience, the role of thymectomy in improving long term outcome in nonthymomatous myasthenia gravis remains controversial. PMID- 11437684 TI - What are the alternatives to increasing inhaled corticosteroids for the long term control of asthma? AB - The Global Initiative for Asthma (GINA) guidelines stated the therapeutic goals for the management of asthma and, through a stepwise approach to treatment, defined the various grades of asthma severity and the therapeutic options available to the clinician at each step. This article considers the options at step 3; the management of a patient with poorly controlled asthma who is already taking low-dose inhaled corticosteroids. Before considering a change in therapy, the clinician should rule out alternative diagnoses, confirm compliance with treatment, explore potential exacerbants in the patient's environment and, where possible, remove them. If a change in medication is necessary, the choice of drug will depend on the therapeutic goal that needs to be achieved. If the most important goal is the control of symptoms and optimisation of lung function, most studies support the addition of a long-acting beta(2)-agonist to low dose inhaled corticosteroids. If recurrent severe exacerbations are a major feature of the poor control, increasing the dosage of inhaled corticosteroids may be most effective. The addition of a leukotriene antagonist may be the best choice if exercise-induced symptoms are prominent or in the setting of aspirin-sensitive asthma. General recommendations supported by the findings of large therapeutic trials do not allow for significant variability in the individual response to a particular drug. Receptor polymorphisms have recently been discovered that may account for variability in the response to beta(2)-agonists and leukotriene receptor antagonists. However, until more is known about the reasons behind this variability, a therapeutic trial may be the most effective way of determining the best drug for an individual patient. One of the key developments in asthma over the past decade has been the acceptance of the concept of asthma as a chronic inflammatory disorder of the airways. However, the long term significance of this inflammation is not clear and the importance of control of inflammation beyond the suppression of symptoms, reduction of exacerbation frequency and the optimisation of lung function has not been established. PMID- 11437685 TI - An infectious basis for multiple sclerosis: perspectives on the role of Chlamydia pneumoniae and other agents. AB - The aetiology of multiple sclerosis (MS) remains unknown. Epidemiological, clinical and pathological data support the theory that MS is a complex disease/syndrome with many factors affecting its development and progression. It may be appropriate to regard MS as a syndrome with differing clinical and pathological features occurring along a spectrum. Patients with MS are more likely to have an affected relative than are individuals without MS, which suggests that there is a genetic component to this illness. Despite this genetic susceptibility, 85% of MS patients do not have an affected relative and only 1 in 3 monozygotic (identical) twins develops MS if the other twin already has it. These data strongly suggest that environmental factors influence the development of MS. Many putative infectious agents have been proposed to be involved in the aetiology of MS. Although research into identifying MS-causative agents dates back for more than 5 decades, no agent has yet emerged with any consensus as the cause of MS. This controversy is due to a number of factors, including lack of specificity of an agent to MS, lack of reproducibility in other laboratories, inappropriate controls, laboratory contamination and lack of a standard and easily reproducible assay system. Chlamydia pneumoniae is a recently described pathogen that may have a role in the pathogenesis of MS. C. pneumoniae is an intracellular bacterial organism that is infectious to humans. It has recently been detected in the cerebrospinal fluid (CSF) of MS patients but not in that of patients with other neurological diseases. There is also a case report of a patient with CNS C. pneumoniae infection and rapidly progressive MS responding to antimicrobial therapy directed against this pathogen. An association between C. pneumoniae in the CSF and MS is now apparent, but its role in the development of MS remains unknown. Further work exploring the role of C. pneumoniae in inflammatory demyelination is required. This may be accomplished either by developing an animal model or in a therapeutic trial in patients with MS. PMID- 11437686 TI - Immunotherapy in patients with systemic mycoses: a promising adjunct. AB - Evidence from several in vitro and animal model studies suggests a modulatory role of haemopoietic, T(H)1 and T(H)2 cytokines in host defence against fungi, and highlights their potential utility as adjunctive therapy for management of systemic mycoses (SM). However, there are limited clinical data to support the use of cytokines in prevention and treatment of SM. Thus, at present no adjunctive treatment is justified for routine use in all patients. Potential application of these immunomodulatory agents include the use of granulocyte macrophage colony-stimulating factor or macrophage colony-stimulating factor in the management of mycoses in neutropenic patients with myelogenous leukaemia or bone marrow transplantation. Interferon-gamma may have a useful role against aspergillosis in patients with chronic granulomatous disease. Granulocyte colony stimulating factor-elicited white blood cell transfusions may be life saving to patients with refractory SM. Better understanding of synergy between cytokines and specific antifungals may provide powerful tools for managing these serious infections. PMID- 11437687 TI - Immunoliposomes: a promising approach to targeting cancer therapy. AB - Immunoliposomes (antibody-coupled liposomes) have been regarded as very attractive drug-targeting systems for chemotherapeutic cancer treatment. Fundamental problems regarding immunoliposome preparation and application such as antibody coupling and immunoliposome stability and pharmacokinetics have been overcome during the last decade. Therefore, several promising studies on tumour targeting have been described in recent years. Adding to existing reviews on liposomal drug delivery, this article focuses on immunoliposome tumour targeting and summarises various experiments of immunoliposome application in vitro and in vivo with respect to structural liposomal parameters, therapeutic potential and the requirements of the target sites. New therapeutic trends related to immunoliposomes are also considered. Remaining problems in immunoliposome application, especially immunological aspects, are discussed, as are strategies that might help to overcome these obstacles. PMID- 11437688 TI - Combination therapy with interferon-alpha and ribavirin for hepatitis C: practical treatment issues. AB - Combination therapy with ribavirin and interferon (IFN)-alpha for 6 to 12 months is currently the treatment of choice for chronic hepatitis C infection. The overall sustained response rate to treatment, defined as loss of hepatitis C virus (HCV) from serum 6 months after completion of treatment, is 40%. The indications for treatment are serum HCV RNA positivity, abnormal serum transaminases and the presence of portal fibrosis and/or moderate/severe inflammation. Response rates are lower in genotype 1 than in genotype 2 or 3 and in the presence of a high viral load. Anaemia is the most common adverse event and is due to ribavirin; neuropsychiatric adverse effects due to IFNalpha lead to premature cessation of therapy in 10 to 20% of patients. The current recommended dose of interferon is 3MU given subcutaneously 3 times a week. However, it is likely that longer-acting pegylated interferons, which may be more effective and can be administered once weekly, will in the future replace currently used IFNalpha. PMID- 11437689 TI - Comparative efficacy of inhaled corticosteroids and antileukotriene drugs in asthma. AB - Asthma is an inflammatory disease of the airways that is best treated by minimising exposure to factors that provoke the inflammation (e.g. allergens) and by administering drugs that reduce the inflammatory response. The cornerstone of asthma treatment is inhaled corticosteroids. Their effectiveness is a result of their potent and broad anti-inflammatory properties. Antileukotriene drugs (leukotriene modifiers) provide an alternative and novel approach to the treatment of asthma. The novelty of these new compounds is that their effectiveness is firmly based on the pathophysiology of asthma, specifically the role played by the cysteinyl leukotrienes. At the same time, the availability of the antileukotriene drugs has stirred debate over when they should be used and how they compare to inhaled corticosteroids. Although the answers are not fully known at this time, the currently available published and presented data are adequate for us to draw some conclusions about their relative effectiveness and role in asthma treatment. The antileukotriene drugs are more effective than placebo, but they are not as effective as inhaled corticosteroids in improving lung function [measured as the forced expiratory volume in 1 second (FEV(1)) or peak expiratory flow rate (PEFR)], reducing beta(2)-agonist use, and decreasing symptom-free days. In contrast, they may have similar beneficial effects on reducing asthma exacerbations and decreasing peripheral blood eosinophil counts. In the absence of knowing a priori the response of an individual patient to treatment with either therapy, the data favour initiating treatment with an inhaled corticosteroid. However, for patients with mild to moderate disease there are a number of circumstances that support using an antileukotriene drug first. A few examples are aspirin intolerance, predominantly exercise-induced symptoms and problems with using an inhaler or the adverse effects of inhaled corticosteroids such as dysphonia and thrush. For patients with more severe disease, inhaled corticosteroids remain the treatment of choice. Antileukotriene drugs should be considered as add-on therapy, especially in view of their possible complementary effects on reducing airway inflammation. PMID- 11437690 TI - Tumour necrosis factor-alpha blockers in rheumatoid arthritis: review of the clinical experience. AB - Tumour necrosis factor (TNF)-alpha has been found to play a central role in the pathogenesis of rheumatoid arthritis, leading to development of novel drug therapies that neutralise the deleterious effects of this cytokine. This new concept of immunobiological treatment of rheumatoid arthritis has yielded successful results. Although the 2 currently available TNFalpha blockers, infliximab and etanercept, differ in structure, mechanism of action and pharmacokinetics, they have provided similar benefits both in clinical improvement and in slowing and even arresting the progression of radiographic damage. This therapeutic response seems to be unequalled by "conventional" treatments in rheumatoid arthritis, and is incontestably a turning point in the therapeutic management of this disease. PMID- 11437691 TI - Risk factors for and management of post-transplantation cardiovascular disease. AB - The mortality rates due to cardiovascular disease (CVD) in transplant recipients are greater than in the general population. CVD is a major cause of both graft loss and patient death in renal transplant recipients, and improving cardiovascular health in transplant recipients will presumably help to extend both patient and graft survival. Further studies are needed to better evaluate the effectiveness of risk modification on subsequent CVD morbidity and mortality. There is no reason to consider risk factors for CVD such as hyperlipidaemia, hypertension and diabetes mellitus in transplant recipients differently from in the general population. In addition, there are specific transplantation risk factors such as acute rejection episodes and the use of immunosuppressive drugs. It is obvious that several of the immunosuppressive agents used today have disadvantageous influences on risk factors for CVD such as hyperlipidaemia, hypertension and post-transplantation diabetes mellitus (PTDM), but the relative importance of immunosuppressant-induced increases in these risk factors is basically unknown. This may be a strong argument for the selective use and individual tailoring of immunosuppressive agents based upon the risk factor profile of the patient, without jeopardising the function of the graft. Hyperlipidaemia is common after transplantation, and immunosuppression with corticosteroids, cyclosporin, or sirolimus (rapamycin) causes different types of post-transplantation hyperlipidaemia. However, to date, no studies have demonstrated that lipid lowering strategies significantly reduce CVD morbidity or mortality and improve allograft survival in transplant recipients. Several studies using preventive or interventional approaches are ongoing and will be reported in the near future. Post-transplantation hypertension appears to be a major risk factor determining graft and patient survival, and immunosuppressive agents have different effects on hypertension. Controlled studies support the opinion that post-transplantation hypertension must be treated as strictly as in a population with essential hypertension, diabetes mellitus, or chronic renal failure. As increasing numbers of immunosuppressive agents become available for use, we may be in a better position to tailor immunosuppressive therapy to the individual patient, avoiding the use of diabetogenic drugs, drug combinations, or inappropriate doses in patients susceptible to PTDM. Multiple acute rejection episodes have also been demonstrated to be a risk factor for CVD - a strong argument for the use of immunosuppressive drugs to reduce acute rejection. Until we have a better understanding from ongoing landmark studies on the management of CVD, presently available therapy to reduce risk factors needs to be used together with individual tailoring of immunosuppressive therapy with the aim of reducing CVD in these patients. PMID- 11437692 TI - C2 monitoring strategy for optimising cyclosporin immunosuppression from the Neoral formulation. AB - Profiling of absorption of cyclosporin microemulsion (Neoral) is a concept in therapeutic drug monitoring (TDM) designed to further optimise the clinical benefits of this formulation in transplant recipients. A single blood concentration measurement 2 hours after Neoral administration (C2) has been shown in both liver and kidney transplant recipients to be a significantly more accurate predictor of drug exposure than trough concentrations (C0), and its use results in a reduction in the incidence and severity of cellular rejection. In a prospective trial in de novo renal transplant recipients, patients who achieved target concentrations for area under the concentration-time curve over the first 4 hours postdose (AUC(0-4h)) of 4500 to 5500 ng. h/ml within 5 days of transplantation had a 7% incidence of histological acute rejection, compared with 37% rejection in those patients who did not achieve this target level. Of the single sampling points, C2 correlates best with AUC(0-4h) (r2 = 0.86); C(0) had the poorest correlation. In an international study in 21 centres examining the absorption profiling, C2 samples were the most accurate predictors of AUC(0-4h) and freedom from rejection. In liver transplant recipients receiving Neoral based maintenance immunosuppression, adoption of Neoral C2 monitoring identifies patients who are both over- and under-dosed, which is not distinguished by C0 measurements. Further adjustment of C2 to recommended targets, even at 5 and 10 years after transplantation, results in reduction in nephrotoxicity without exposing the patient to the risk of rejection. In summary, despite a level of simplicity comparable to C0 measurement, Neoral absorption profiling, and specifically C2 measurement, is a much more sensitive approach to assessing the pharmacokinetics and predicting the clinical effect of this formulation in the individual patient, with a consequent marked reduction in the incidence of acute cellular rejection and improved long term graft function. PMID- 11437693 TI - Pathophysiology of immune-mediated (type 1) diabetes mellitus: potential for immunotherapy. AB - Type 1 diabetes mellitus is a chronic T cell-mediated disease resulting from autoimmune destruction of pancreatic beta-cells. This process leads to progressive and irreversible failure of insulin secretion. Development of the disease involves both genetic and environmental factors. Genetic predisposition is mainly connected with the human leucocyte antigen (HLA) region, which encodes structures responsible for antigen presentation. A comprehensive molecular understanding of the pathogenesis of the disease is essential for the design of rational and well tolerated means of prevention. This paper describes recent experimental and clinical findings and elucidates the current possibilities for immunotherapy of type 1 diabetes. The nature of breakdown of self-tolerance and the mechanisms involved in its recovery are discussed. PMID- 11437694 TI - Prophylaxis and treatment of influenza virus infection. AB - Influenza virus infections remain an important cause of morbidity and mortality. Furthermore, a recurrence of pandemic influenza remains a real possibility. There are now effective ways to both prevent and treat influenza. Prevention of infection is most effectively accomplished by vaccination. Vaccination with the inactivated, intramuscular influenza vaccine has been clearly demonstrated to reduce serious morbidity and mortality associated with influenza infection, especially in groups of patients at high risk (e.g. the elderly). However, the inactivated, intramuscular vaccine does not strongly induce cell-mediated or mucosal immune responses, and protection induced by the vaccine is highly strain specific. Live, attenuated influenza vaccines administered intranasally have been studied in clinical trials and shown to elicit stronger mucosal and cell-mediated immune responses. Live, attenuated vaccines appear to be more effective for inducing protective immunity in children or the elderly than inactivated, intramuscular vaccines. Additionally, novel vaccine methodologies employing conserved components of influenza virus or viral DNA are being developed. Preclinical studies suggest that these approaches may lead to methods of vaccination that could induce immunity against diverse strains or subtypes of influenza. Because of the limitations of vaccination, antiviral therapy continues to play an important role in the control of influenza. Two major classes of antivirals have demonstrated ability to prevent or treat influenza in clinical trials: the adamantanes and the neuraminidase inhibitors. The adamantanes (amantadine and rimantadine) have been in use for many years. They inhibit viral uncoating by blocking the proton channel activity of the influenza A viral M2 protein. Limitations of the adamantanes include lack of activity against influenza B, toxicity (especially in the elderly), and the rapid development of resistance. The neuraminidase inhibitors were designed to interfere with the conserved sialic acid binding site of the viral neuraminidase and act against both influenza A and B with a high degree of specificity when administered by the oral (oseltamivir) or inhaled (zanamivir) route. The neuraminidase inhibitors have relatively low toxicity, and viral resistance to these inhibitors appears to be uncommon. Additional novel antivirals that target other phases of the life cycle of influenza are in preclinical development. For example, recombinant collectins inhibit replication of influenza by binding to the viral haemagglutinin as well as altering phagocyte responses to the virus. Recombinant techniques have been used for generation of antiviral proteins (e.g. modified collectins) or oligonucleotides. Greater understanding of the biology of influenza viruses has already resulted in significant advances in the management of this important pathogen. Further advances in vaccination and antiviral therapy of influenza should remain a high priority. PMID- 11437695 TI - Immunological aspects of alzheimer's disease: therapeutic implications. AB - Alzheimer's disease (AD) is a chronic neurodegenerative disease causing progressive impairment of memory and cognitive function. The amyloid cascade hypothesis suggests that mismetabolism of the beta-amyloid (A beta) precursor protein (APP) followed by subsequent formation of non-fibrillar and fibrillar A beta deposits leads to glial activation and eventually to neurotoxicity, causing cognitive impairment. Several lines of evidence indicate that an inflammatory process contributes to the pathology of AD. First, inflammatory proteins have been identified as being associated with neuritic plaques and in glial cells surrounding these plaques. Second, certain polymorphisms of acute-phase proteins and cytokines associated with AD plaques increase the risk or predispose for earlier onset of developing AD. Third, epidemiological studies indicate that anti inflammatory drugs can retard the development of AD. Several steps in the pathological cascade of AD have been identified as possible targets for actions of nonsteroidal anti-inflammatory drugs. For instance, microglia are considered a target because this cell type is closely involved in AD pathology through secretion of neurotoxic substances and by modulating a positive feedback loop of the inflammatory mechanism that may be involved in the pathological cascade in AD. On the basis of studies in APP transgenic mice, immunisation with A beta was recently suggested as a novel immunological approach for the treatment of AD. Immunisation elicits A beta-specific antibodies that could affect several early steps of the amyloid-driven cascade. Antibodies could prevent A beta from aggregating into fibrils and accelerate clearance of A beta by stimulating its removal by microglial cells. This review outlines the pathological and genetic evidence that an inflammatory mechanism is involved in AD and the therapeutic approaches based on inhibition or mediation of inflammation. PMID- 11437696 TI - Tolerability of aminosalicylates in inflammatory bowel disease. AB - Since its synthesis in the 1930s and subsequent introduction, sulfasalazine has been an effective treatment for inflammatory bowel disease. However, up to one third of patients are unable to take the drug because of severe intolerance. The finding in 1977 that the anticolitic effect of sulfasalazine lay in its 5 aminosalicylic [(5-ASA); mesalazine] moiety led to the development of new generations of 5-ASA agents. These new agents include a slow continuous release formulation, pH-dependent release formulations, formulations using alternative carrier molecules and rectally administered formulations. Newer 5-ASA formulations are more effective than placebo in maintaining remission of ulcerative colitis. They have also been used for the treatment of active Crohn's disease as well as maintenance treatment of ileocolonic Crohn's disease, although their role in isolated small bowel disease is controversial. In general terms, all of the newer 5-ASA preparations are much better tolerated than sulfasalazine. The use of standard dosages of mesalazine in pregnancy appears to be tolerated; however, continuing surveillance of pregnancy outcome is recommended. While there is evidence that mesalazine can cause nephrotoxic reactions, these reactions can occur with all 5-ASA-containing preparations, particularly in individuals with existing renal disease. Blood dyscrasias can also occur with all aminosalicylates. PMID- 11437714 TI - Laparoscopic ablation is not necessary for minimal or mild lesions in endometriosis associated subfertility. PMID- 11437715 TI - Serum leptin levels in pregnant women with type 1 diabetes mellitus. AB - BACKGROUND: Leptin is an important weight regulator and during pregnancy leptin is not only synthesized in adipose tissue but also in the placenta. AIM: To examine changes in serum leptin levels in women with type 1 diabetes mellitus during pregnancy and post delivery in relation to concomitant changes in maternal body weight, birth weight, glycemic control, and blood pressure. METHODS: Non fasting serum leptin from 45 women with type 1 diabetes mellitus were studied consecutively throughout pregnancy and 3 months post partum. RESULTS: Serum leptin was positively associated with HbA1c in week 18, 22 and 30 (r=0.38, 0.41, and 0.54, respectively, p<0.05, adjusted for body weight). Moreover, serum leptin correlated positively with maternal body weight and BMI (0.4525 kg/m2), the changes during pregnancy and the level of serum leptin were significantly greater compared to lean women (p<0.05). The women with low ambulatory blood pressure (lower tertile, mean arterial blood pressure <83.4 mmHg) showed the lowest level of serum leptin throughout pregnancy and it changed significantly differently from the women with higher blood pressure (p<0.05). CONCLUSION: Changes in serum leptin levels of pregnant women with type 1 diabetes mellitus were associated with parallel changes in maternal body weight and glycemic control. Women with low blood pressure had the lowest serum leptin levels throughout pregnancy. PMID- 11437716 TI - Levels of dimethylarginines and cytokines in mild and severe preeclampsia. AB - BACKGROUND: The objectives were 1. to evaluate if the endogenous nitric oxide synthase inhibitor asymmetric dimethylarginine was altered in mild and severe forms of preeclampsia, and 2. to assess the relationship between dimethylarginines and the cytokine response in preeclampsia. METHODS: Asymmetric and symmetric dimethylarginine were measured with high performance liquid chromatography in women with mild (n=13) and severe (n=32) preeclampsia and in normotensive pregnant controls (n=20). Interleukin-4, -6, -8, -10 and tumor necrosis factor-alpha were analyzed by immunoassays in women with mild (n=8) and severe (n=17) preeclampsia and in normotensive pregnant controls (n=14). The Mann Whitney U-test and Spearman Rank test were used for statistical analysis. RESULTS: The plasma levels of dimethylarginine were increased in preeclamptic subjects. The elevation of symmetric dimethylarginine was more pronounced than that of asymmetric dimethylarginine. The control levels of interleukin-6, -8 and 10 were significantly higher at term than at gestational week 32-36. Interleukin 6 and -8 were significantly elevated in subjects with severe, but not mild, preeclampsia, whereas TNF-alpha and IL-10 were not significantly altered. Symmetric dimethylarginine levels correlated significantly with arterial blood pressure and serum levels of creatinine and uric acid. Dimethylarginine levels in plasma were, however, not related to the cytokine response. CONCLUSIONS: Plasma concentrations of both asymmetric and symmetric dimethylarginine were significantly elevated both in mild and severe preeclampsia. Symmetric, but not asymmetric, dimethylarginine correlated to the severity of the condition. Plasma levels of interleukin-6 and -8 were also elevated in severe preeclampsia but no direct correlations were found between these cytokines and dimethylarginines. PMID- 11437717 TI - Can uterine artery Doppler velocimetry predict adverse pregnancy outcome in women with antiphospholipid syndrome? PMID- 11437718 TI - Volume sonography of the pelvic floor in nulliparous women and after elective cesarean section. AB - OBJECTIVE: To describe the sonographic appearance of the pelvic floor which has not been stressed by forces of labor or vaginal delivery in pregnant women and after childbirth. SUBJECTS AND METHODS: In a prospective observational study 14 nulliparous women during first trimester pregnancy and 26 primiparous women after elective cesarean were examined within the first week postpartum. The integrity of the internal anal sphincter expressed as the ratio between the anterior and the posterior internal anal sphincter muscle thickness (a/p-ratio), asymmetry of the levator ani muscle and the paraurethral fixation of the lateral vaginal edges at the arcus tendineus were assessed using volume sonography. RESULTS: The a/p ratio in nulliparous women was significantly higher (p<0.01) than after elective cesarean section. Asymmetry of the levator ani muscle did not differ between both groups but was more frequently in the nulliparous patients with an odds-ratio of 1.16 (CI 0.74-1.82). In both groups of women the paraurethral fixation of the lateral vaginal edges were above the suburethral level of the vagina. CONCLUSIONS: This study gives sonographic features of the pelvic floor in nulliparous women and in primigravidae after elective cesarean section. Data from the post-cesarean group can serve as reference values for further studies evaluating pelvic floor damage after various modi of vaginal delivery. PMID- 11437719 TI - Transvaginal ultrasonography sonohysterography and operative hysteroscopy for the evaluation of abnormal uterine bleeding. AB - OBJECTIVE: To evaluate the diagnostic accuracy of transvaginal ultrasonography, sonohysterography and hysteroscopy in patients presenting with abnormal uterine bleeding. DESIGN: Prospective, blind, comparative study. SETTING: Outpatient clinic and day surgery unit of a district hospital specializing in operative hysteroscopy. PATIENTS: One hundred consecutive patients referred with abnormal uterine bleeding (AUB). INTERVENTIONS: Transvaginal ultrasonography and sonohysterography were performed in the outpatient clinic. Within 1 week, all patients underwent operative hysteroscopy. MAIN OUTCOME MEASURES: All diagnoses established by transvaginal ultrasonography, and sonohysterography were compared to the appearance of the cavity using hysteroscopy as well as the histological diagnosis from removed tissues. RESULTS: In 88 patients information suitable for analysis was obtained by all methods. The detection rate of focal intrauterine pathology using sonohysterography was (94.1%), but was significantly lower with transvaginal ultrasonography (23.5%). In about 75% of all cases none of the methods used was able to correctly detect endometrial hyperplasia. CONCLUSIONS: Sonohysterography was significantly better than transvaginal ultrasonography in detecting focal intrauterine pathology. Visual examination at operative hysteroscopy yielded no additional information to the detection or exclusion of focal lesions than was obtained at outpatient sonohysterography. All methods performed equally in assessing endometrial pathology. We conclude that sonohysterography may replace diagnostic hysteroscopy in many patients with AUB. PMID- 11437720 TI - Increased acceptance of HRT and improved level of information: a change in Norwegian women's opinion from 1990 to 1997. AB - AIM: To investigate the development in women's attitude to hormone replacement therapy, and the change in views concerning positive and negative effects. METHODS: National cohort study based on telephone interviews in 1990 and 1997, of women randomly sampled from the Norwegian telephone book (n=1019 and 1003). The same questions were asked in both studies. RESULTS: In 1997, more women stated radio and television, doctors, friends and relatives as sources of information. The general attitude to hormone replacement therapy had become significantly more positive, and the respondents reported a higher level of information than in 1990. However, some misunderstandings about estrogen therapy still remain; the proportion of women who believed that estrogen treatment reduced the risk for breast cancer increased from 4% in 1990 to 15% in 1997. About one third of women aged 60 or more are current or past users. More than 60% of the women in the youngest age group state that they will consider use of estrogen replacement therapy. About four-fifths of the women in 1997 stated their willingness to use long term estrogen therapy if osteoporosis could be prevented. CONCLUSIONS: Estrogen replacement therapy has become a part of the public debate. A high degree of willingness to use estrogen is demonstrated. The self reported level of information is higher and women are generally more correctly informed about positive and negative effects. PMID- 11437721 TI - Changes in quality of life after hormonal treatment of endometriosis. AB - BACKGROUND: To assess whether hormonal treatment of endometriosis improves quality of life for women with endometriosis. METHODS: In a prospective, randomized, double-blind, double-dummy study on 48 women with verified endometriosis, the pain pattern and quality of life were registered before, during and after treatment with nafarelin or medroxyprogesterone acetate. The distribution of the studied parameters were tested by means of a Skewness test. ANOVA analysis was used for normally distributed variables and Friedman's analysis and Mann-Whitney U-test for non normally distributed variables. RESULTS: There was a difference between the 30 women who participated all through the study and the 18 who dropped out. It was noticeable that anxiety-depression and sleep disturbances were significantly more common among the drop outs. There was a significant reduction in symptom score during the study, without any significant difference between the treatment groups. The sleep disturbances and anxiety-depression score improved significantly from before treatment to the end of the follow-up, but the anxiety-depression score increased during the nafarelin treatment period. There was a statistically significant improvement of paid working life in the nafarelin treated group. All the other psycho-social parameters as well as emotional balance improved during the study period without difference between groups. CONCLUSION: When hormonal treatment is planned it is very important to take into consideration previous psychosocial experiences of the patient. Factors of importance for quality of life such as sleep disturbances and anxiety-depression improved significantly after treatment with nafarelin or medroxyprogesterone acetate. PMID- 11437722 TI - The expression of cyclooxygenase 2 and inducible nitric oxide synthase indicates no active inflammation in vulvar vestibulitis. AB - BACKGROUND: Although women with vulvar vestibulitis syndrome have principal symptoms of inflammation such as local erythema and pain in the mucosa around the vaginal introitus, it is not clear if vestibulitis is an inflammatory condition. Cyclooxygenase 2 and inducible nitric oxide synthase are known to be upregulated during inflammation. The aim of the present study was to analyze the expression of these enzymes in the vestibular mucosa in order to evaluate the inflammatory activity in the tissue. METHODS: Ten women fulfilling Friedrich's criteria of vulvar vestibulitis syndrome and ten control subjects were included in the study. Punch biopsies were obtained from the vestibular mucosa for analysis of cyclooxygenas 2 and inducible nitric oxide synthase, using indirect immunohistochemistry and Western dot-blot analyses. RESULTS: Both methods used showed low expression of cyclooxygenas 2 and inducible nitric oxide synthase in the vestibular mucosa of all women. There was no difference observed between the groups. CONCLUSIONS: There is a low expression of the inflammatory markers cyclooxygenas 2 and inducible nitric oxide synthase in the vestibular mucosa of women with vulvar vestibulitis syndrome as well as in healthy control subjects. The results indicate no active inflammation present and imply that topical corticosteroids in the treatment of vulvar vestibulitis are unfounded. PMID- 11437723 TI - Can the endometrial thickness as measured by trans-vaginal sonography be used to exclude polyps or hyperplasia in pre-menopausal patients with abnormal uterine bleeding? AB - BACKGROUND: To combine findings of normal mid-line echoes from TVS (trans-vaginal sonography) with cut-off levels for endometrial thickness, in order to examine the associated risk of abnormalities in the uterine cavity. DESIGN AND SETTING: Two center prospective study at a university clinic and a central hospital in Denmark. PARTICIPANTS: Three hundred and fifty-five pre-menopausal patients with abnormal uterine bleeding, and indications for endometrial sampling or surgery. INTERVENTIONS: The thickness of the endometrium was measured, and the mid-line echoes were evaluated using TVS. The findings from the endometrial sampling, combined with the evaluation of the uterine cavity using operative hysteroscopy (115), hysterectomy (74) or HSE (hysterosonographic examination) (166), were used as the true values. RESULTS: The mean (+/-s.d.) endometrial thickness was significantly different in patients with hyperplasia 11.5 mm (+/-5.0), polyps 11.8 mm (+/-5.1), sub-mucous myomas 7.1 mm (+/-3.4) and in patients without these abnormalities 8.37 (+/-3.9) (p<0.001). Hyperplasia and/or polyps were present in 20% of all patients, and in 8% of 143 patients with an endometrial thickness of < or =7 mm. This proportion did not decrease with lower cut-off levels for endometrial thickness. Receiver operating characteristic (ROC) curves were not optimal for excluding hyperplasia or polyps by endometrial thickness. In 173 cases with a distinct, regular midline echo without echo-dense foci in TVS the proportion of patients with abnormalities was 16% (11-23). This proportion did not decrease with cut-off levels for endometrial thickness. CONCLUSIONS: Using TVS, low levels of endometrial thickness reduced the possibility of abnormalities such as polyps and hyperplasia, but did not exclude them. Low cut-off levels for endometrial thickness did not increase the diagnostic performance in cases with normal sonograms. PMID- 11437724 TI - Y-chromosomal microdeletions among infertile Finnish men. AB - BACKGROUND: Microdeletions in the Y-chromosome are known to cause a significant proportion of azoo- and oligozoospermia in men. The reported frequency of deletions varies greatly between the studies. Probable reasons for this variation are different selection criteria and number of patients included, and possibly also methodological aspects, whereas the contribution of environmental and genetic factors is not known. The aim of this study was to determine the incidence of Y-chromosome microdeletions among infertile Finnish men. METHODS: Two hundred and one men showing azoospermia (n=68) or severe oligozoospermia (n=133) were included. Multiplex polymerase chain reaction method was used to amplify specific sequence tagged sites (STS) along the Y chromosome. RESULTS: Microdeletions were observed in 18 men (9%), of whom four were azoospermic and 14 oligozoospermic. CONCLUSIONS: The incidence of Y-deletions in the study population of infertile Finnish men falls within the range published in other countries. PMID- 11437725 TI - Cervical pregnancy with hydatidiform mole. PMID- 11437726 TI - Papillary adenocarcinoma of the ovary presenting in a PAP-smear. PMID- 11437727 TI - Ovarian pregnancy: a case report. PMID- 11437728 TI - Cervical pregnancy and therapeutic options. PMID- 11437729 TI - Giant ovarian tumor complicating late pregnancy: A case report. PMID- 11437730 TI - Conservative treatment for a noncommunicating rudimentary uterine horn. PMID- 11437731 TI - Bone metastasis as the presenting complaint in ovarian carcinoma. PMID- 11437732 TI - Intrauterine pregnancy after thermal balloon ablation. PMID- 11437733 TI - Metastatic placental site trophoblastic tumors: clinical aspects of two unique cases. PMID- 11437734 TI - Glutathione deficiency intensifies ischaemia-reperfusion induced cardiac dysfunction and oxidative stress. AB - The efficacy of glutathione (GSH) in protecting ischaemia-reperfusion (I-R) induced cardiac dysfunction and myocardial oxidative stress was studied in open chest, stunned rat heart model. Female Sprague-Dawley rats were randomly divided into three experimental groups: (1) GSH-depletion, by injection of buthionine sulphoxamine (BSO, 4 mmol kg(-1), i.p.) 24 h prior to I-R, (2) BSO injection (4 mmol kg(-1), i.p.) in conjunction with acivicin (AT125, 0.05 mmol kg(-1), i.v.) infusion 1 h prior to I-R, and (3) control (C), receiving saline treatment. Each group was further divided into I-R, with surgical occlusion of the main left coronary artery (LCA) for 30 min followed by 20 min reperfusion, and sham. Myocardial GSH content and GSH : glutathione disulphide (GSSG) ratio were decreased by approximately 50% (P < 0.01) in both BSO and BSO + AT125 vs. C. Ischaemia-reperfusion suppressed GSH in both left and right ventricles of C (P < 0.01) and left ventricles of BSO and BSO + AT125 (P < 0.05). Contractility (+dP/dt and -dP/dt) in C heart decreased 55% (P < 0.01) after I and recovered 90% after I-R, whereas +/-dP/dt in BSO decreased 57% (P < 0.01) with ischaemia and recovered 76 and 84% (P < 0.05), respectively, after I-R. For BSO + AT125, +/ dP/dt were 64 and 76% (P < 0.01) lower after ischaemia, and recovered only 67 and 61% (P < 0.01) after I-R. Left ventricular systolic pressure in C, BSO and BSO + AT125 reached 95 (P > 0.05) 87 and 82% (P < 0.05) of their respective sham values after I-R. Rate-pressure double product was 11% (P > 0.05) and 25% (P < 0.05) lower in BSO and BSO + AT125, compared with Saline, respectively. BSO and BSO + AT125 rats demonstrated significantly lower liver GSH and heart Mn superoxide dismutase activity than C rats after I-R. These data indicate that GSH depletion by inhibition of its synthesis and transport can exacerbate cardiac dysfunction inflicted by in vivo I-R. Part of the aetiology may involve impaired myocardial antioxidant defenses and whole-body GSH homeostasis. PMID- 11437735 TI - Effects of right and left vagal stimulation on left ventricular acetylcholine levels in the cat. AB - To test the effectiveness of, and the interactions between, right and left vagal stimulation on left ventricular acetylcholine (ACh) levels, we applied the dialysis technique to the heart of anaesthetized cats. Dialysis probes were implanted in the left ventricular myocardium and perfused with Krebs-Henseleit buffer containing eserine. Dialysate ACh content was measured as an index of ACh release from post-ganglionic vagal nerve terminals in the left ventricular myocardium. We electrically stimulated the right and left cervical vagal nerves separately or together and investigated the dialysate ACh response. In two different regions of the left ventricle, substantial dialysate ACh responses were observed by the stimulation (20 Hz) of both right and left cervical vagal nerves. At stimulation frequencies of both 10 and 20 Hz, the dialysate ACh response to the bilateral vagal stimulation was almost algebraically the calculated sum of the individual dialysate ACh responses to unilateral vagal stimulation. In conclusion, ACh levels in the left ventricle are affected by both right and left vagal nerves and show little evidence of interactions between right and left vagal nerves at the level of the cardiac ganglia. PMID- 11437736 TI - Prolactin and lipopolysaccharide treatment increased apoptosis and atresia in rat ovarian follicles. AB - Follicular atresia is associated with the presence of increased macrophages within the follicle. What is not known is whether, in the adult rat, macrophages are instrumental in inducing apoptosis and/or atresia or whether they are simply secondary to a hormonally mediated event. As prolactin is an immunoreactive hormone and stimulates the expression of monocyte chemoattractant, the present experiments compared the effects of prolactin treatment with that of an immune challenge with lipopolysaccharide (LPS) on the invasion of macrophages into the follicular and luteal compartments of the ovary and the occurrence of apoptosis/atresia in relation to macrophage invasion. Rats were treated for 3 days with either prolactin or LPS and ovaries obtained at pro-oestrus or oestrus. Prolactin and LPS increased the number of atretic vs. healthy follicles (P < 0.008, chi2) in pro-oestrus ovaries and increased the mean number of apoptotic cells and macrophages (P < 0.05 for some groups). Macrophages were typically observed in the thecal layer, apoptotic cells in the granulosa cell layer, although 84% follicles which had macrophages within the granulosa cell layer also contained relatively high numbers of apoptotic nuclei. Prolactin and LPS treatment in vivo reduced the progesterone response to follicle stimulating hormone (FSH) (P < 0.001) in cultures of ovarian dispersates but did not inhibit the response to forskolin. In contrast, prolactin or LPS added in vitro to the cultures inhibited the progesterone response to forskolin. Results show that both prolactin and LPS increase follicular apoptosis and atresia and reduce the progesterone response to FSH. PMID- 11437737 TI - Caffeine stimulates the reverse mode of Na/Ca2+ exchanger in ferret ventricular muscle. AB - This study investigated the effect of caffeine on the sarcolemmal mechanisms involved in intracellular calcium control. Ferret cardiac preparations were treated with ryanodine and thapsigargin in order to eliminate the sarcoplasmic reticulum (SR) function. This treatment abolished caffeine contracture irreversibly in normal solution. The perfusion with K-free medium that blocked the Na+--K+ pump resulted in a recovery of slow relaxing caffeine contractures similar to Na-free contractures. The amplitude of caffeine contractures was dependent on the bathing [caffeine]o and [Ca2+]o. Divalent cations Ni2+ and Cd2+, which have an inhibitory effect on the Na+/Ca2+ exchanger, produced dose dependent inhibition of caffeine responses with apparent Ki of 780 +/- 19 and 132 +/- 5 microM, respectively. Caffeine also caused dose-dependent inhibition of Na free contractures (Ki=4.62 +/- 1.5 mM), and the reduction or removal of [Na+]o exerted an inhibitory effect on caffeine contractures (Ki=73.5 +/- 17.12 mM). These experiments indicate that the increase in resting tension following exposure to caffeine was mediated by Na+/Ca2+ exchanger, which represents an additional element of complexity in caffeine action on cardiac muscle. PMID- 11437738 TI - Brief intense exercise followed by passive recovery modifies the pattern of fuel use in humans during subsequent sustained intermittent exercise. AB - The role of work period duration as the principal factor influencing carbohydrate metabolism during intermittent exercise has been investigated. Fuel oxidation rates and muscle glycogen and free carnitine content were compared between two protocols of sustained intermittent intense exercise with identical treadmill speed and total work duration. In the first experiment subjects (n=6) completed 40 min of intermittent treadmill running involving a work : recovery cycle of 6 : 9 s or 24 : 36 s on separate days. With 24 : 36 s exercise a higher rate of carbohydrate oxidation approached significance (P=0.057), whilst fat oxidation rate was lower (P < or = 0.01) and plasma lactate concentration higher (P < or = 0.01). Muscle glycogen was lower post-exercise with 24 : 36 s (P < or = 0.05). Muscle free carnitine decreased (P < or = 0.05), but there was no difference between protocols. In the second experiment a separate group of subjects (n=5) repeated the intermittent exercise protocols with the addition of a 10-min bout of intense exercise, followed by 43 +/- 5 min passive recovery, prior to sustained (40 min) intermittent exercise. For this experiment the difference in fuel use observed previously between 6 : 9 s and 24 : 36 s was abolished. Carbohydrate and fat oxidation, plasma lactate and muscle glycogen levels were similar in 6 : 9 s and 24 : 36 s. When compared with the first experiment, this result was because of reduced carbohydrate oxidation in 24 : 36 s (P < or = 0.05). There was no difference, and no change, in muscle free carnitine between protocols. A 10-min bout of intense exercise, followed by 43 +/- 5 min of passive recovery, substantially modifies fuel use during subsequent intermittent intense exercise. PMID- 11437739 TI - 2,3-Butanedione monoxime increases speed of relaxation in single muscle fibres of frog. AB - The effects of 2,3-butanedione monoxime (BDM) on intracellular Ca2+ transient and cross-bridge function were studied in frog single fibres from the anterior tibialis muscle of Rana temporaria (sarcomere length, 2.2 microm; temperature, 2 4 degrees C). The fluorescent dye fluo-3 was used to monitor the intracellular free calcium concentration ([Ca2+]i) during isometric contractions. BDM (1-5 mM) reduced the amplitude of the Ca2+ transient during twitches, but this effect was too small to explain the marked inhibition of BDM on twitch force. [Ca2+]i reached at the end of 1-s tetanic stimulation was not significantly affected by BDM (1.0 and 1.8 mM) while the maximum tetanic tension was substantially reduced. The rate of relaxation during isometric tetanus was increased by BDM whereas the rate of decay of the Ca2+ transient was reduced in the presence of BDM. The results strongly suggest that BDM, under the experimental conditions used, mainly affects the contractile machinery resulting in altered performance of the cross bridges. These effects of BDM were evaluated in terms of the cross-bridge model of Huxley (1957) which was fitted to the experimental force-velocity data in the presence and absence of BDM. PMID- 11437740 TI - Force deficits after repeated stretches of activated skeletal muscles in female and male rats. AB - Force deficits after stretches of activated plantar flexor muscles were measured in six male Sprague-Dawley rats (285 +/- 10 g, age 62 +/- 4 days, mean +/- SE) and compared with six age-matched (193 +/- 6 g, age 67 +/- 3 days) and six weight matched female rats (273 +/- 7 g, age 141 +/- 9 days). Twenty stretches, imposed on isometric contractions at 90 degrees (0.2 ms pulse duration, 80 Hz, 5.4 +/- 0.3 V, duty cycle 0.006), were produced by ankle rotation from 90 to 40 degrees. Before the stretch protocol, weight-matched groups had similar isometric forces at an ankle position of 90 degrees at 5, 10, 20, 40, 60 and 80 Hz but forces were lower for age-matched females. For all groups, normalized force-frequency relationships were similar. During the stretch protocol, deficits for isometric force at 90 degrees and peak stretch force at 40 degrees with stretch number were similar for all groups. One hour after the stretches, isometric force deficits at 90 degrees at 40, 60 and 80 Hz were larger for females in weight-matched groups (e.g. 80 Hz, female: 47.8 +/- 1.7%; male: 41.1 +/- 1.7%; P < 0.05), perhaps because of a difference in age (P < 0.05). For age-matched groups, isometric force deficits at 90 degrees were similar at all frequencies. The susceptibility for force deficits by stretches of activated skeletal muscles was not gender dependent for 2-month-old rats. PMID- 11437741 TI - Effects of motility on epithelial transport in the human descending duodenum. AB - The aim of the study was to determine how motility affects the balance between absorptive and secretory ion transport in the proximal human small intestine. Thirty-two healthy subjects and 16 patients (eight with villus atrophy, eight with normal duodenal biopsies) were studied. The absorptive sodium flux was estimated by measurement of bicarbonate absorption with a double-lumen perfusion technique. The secretory chloride flux was calculated from the ratio between the continuously recorded transmural potential difference (PD) and the epithelial parallel resistance, which was measured in forceps biopsies by square-wave current analysis. Graded variations in contraction frequency were obtained by recording during defined time periods before, during and after phase III of the migrating motor complex (MMC). Bicarbonate was absorbed by a process that led to CO2 formation, and both bicarbonate absorption and luminal PCO2 increased with contraction frequency. The motility-related PCO2 rise was reduced in patients with villus atrophy and by removal of bicarbonate from the perfusate. A higher motor activity was also associated with a larger PD (more lumen negative). Both the absorptive and the secretory fluxes were thus enhanced by motility. The estimated absorptive flux was approximately twice as large as the secretory flux during periods of low motor activity, and four times as large during submaximal motor activity. We conclude that motor activity affects both absorptive and secretory mucosal function in a quantifiable manner. Information about the behaviour of the respective linkage functions may make it possible to model the intestinal absorption process in vivo. PMID- 11437742 TI - Administrative aspects in longitudinal studies: how to navigate on a stormy and dangerous ocean? PMID- 11437743 TI - A review of the epidemiological transition in dementia--cross-national comparisons of the indices related to Alzheimer's disease and vascular dementia. AB - OBJECTIVE: To examine temporal changes in the prevalence of dementia and associated factors. METHOD: All publications on the epidemiology of dementia were identified using a medline search for the years 1966-1999. RESULTS: Alzheimer's disease (AD) has become nearly twice as prevalent as vascular dementia (VaD) in Korea, Japan, and China since transition in early 1990s. Prior to this, in the 1980s, VaD was more prevalent than AD in these countries. In Nigeria, the prevalence of dementia was low. Indian studies were contradictory, with both AD and VaD being more prevalent in different studies. American and European studies consistently reported AD to be more prevalent than VaD. CONCLUSION: A theoretical model of transition from low incidence-high mortality society to high incidence high mortality society to low incidence-low mortality society may explain these findings. Rigorous testing in prospective, longitudinal and population-based cross-national studies using culture-fair diagnostic instruments is required. PMID- 11437744 TI - Mental disorders among elderly people in primary care: the Linkoping study. AB - OBJECTIVE: The aims of this study were to describe the prevalence of mental disorders among elderly patients in primary care and to compare diagnoses from psychiatric interview with diagnoses in medical records. METHOD: Patients aged 70 years and above attending a primary care centre (N = 350) were studied using a psychiatric and medical record examination. RESULTS: The prevalence of mental disorder according to the psychiatric interview was 33% (16% dementia, 17% other mental disorders). Only 49% of these had any psychiatric diagnosis in case records and 17-38% received specific treatments. The frequency of psychiatric symptoms among those with no mental disorder was between 1% and 66%. Patients with mental disorders were more often females, had more visits to a doctor, more diagnoses in medical records, and were prescribed more drugs. CONCLUSION: Mental disorders and symptoms are common among the elderly in primary care. More effort should be made to increase the recognition rate. PMID- 11437745 TI - Care utilization and outcome of DSM-III-R major depression in the general population. Results from the Netherlands Mental Health Survey and Incidence Study (NEMESIS). AB - OBJECTIVE: To assess care utilization, individual characteristics and clinical and functional outcomes for various modalities of professional care in people with DSM-III-R major depression. METHOD: Psychiatric diagnoses were determined at baseline and 12-month follow-up in a representative sample (N = 7076) of the Dutch population, using the Composite International Diagnostic Interview (CIDI). RESULTS: A total of 45.3% of the 223 individuals with major depression received professional care in the 12 months between baseline and follow-up, and 42.6% of these were treated with antidepressant medication. Higher level of care was associated with clinical factors and functional limitations. Clinical outcomes were poorly correlated with functional outcomes. Mild to moderate effects in functional outcome were found for all care modalities. CONCLUSION: Outcome of antidepressant treatment can be improved and such treatment should focus on the more severe forms of depression. Functional outcome assessment is recommended in addition to clinical assessment. PMID- 11437746 TI - Dimensionality, responsiveness and standardization of the Bech-Rafaelsen Mania Scale in the ultra-short therapy with antipsychotics in patients with severe manic episodes. AB - OBJECTIVE: Typical antipsychotics have their indication in the ultra-short (first week) treatment of severe episodes of mania. In this setting the Bech-Rafaelsen Mania Scale (MAS) was psychometrically compared with the Clinical Global Impression scale (CGI) to assess its ability to measure response. METHOD: Ratings on patients with marked to severe mania (n = 80) who participated in the clinical trials to evaluate the ultra-short antimanic effect of zuclopenthixol acetate were assessed. The MAS was analysed for internal validity (total score a sufficient statistic) and for external validity. RESULTS: The MAS was shown to have a high internal validity showing onset of action already after days of treatment. After 6 days of treatment 53% of the patients responded according to the MAS but only 30% according to the CGI. The difference was statistically significant. CONCLUSION: The MAS has been found to be a valid scale to measure early onset of action and response in the ultra-short antimanic treatment with typical antipsychotics. PMID- 11437747 TI - Predictive factors of post-discharge follow-up care among adolescent suicide attempters. AB - OBJECTIVE: To study short-term compliance with follow-up care in a sample of adolescent suicide attempters. METHOD: One hundred and sixty-seven adolescents, aged from 13-18 years and hospitalized after a suicide attempt, completed a questionnaire that included the CES-Depression Scale and Zung Anxiety Scale. Physicians assessed the hospital care immediately after the attempt, and post discharge care plans. Three months later, adolescents were contacted by telephone and asked about follow-up care. RESULTS: After 3 months, 91.6% of the adolescents could be contacted: 25.5% never attended any follow-up; 11.1% went only once; 31.4% missed some appointments; and 32.0% went to all their scheduled appointments. Adolescents who complied with follow-up care differed significantly from those who did not: they showed more depression, anxiety and illicit drug use at the time of the attempt; they had more often planned the attempt; they were hospitalized longer; and they met with a psychiatrist more often while hospitalized. Compliance was also better when the follow-up appointment was scheduled before discharge. CONCLUSION: Compliance with post-discharge follow-up care depends upon the adolescent's psychopathology but may also be improved by the type of hospital care and post-discharge plans. PMID- 11437748 TI - Association of personal and familial suicide risk with low serum cholesterol concentration in male lithium patients. AB - OBJECTIVE: We sought to establish whether low cholesterol concentration may be associated with a personal history of attempted suicide or a family history of completed suicide in psychiatric out-patients on maintenance lithium treatment, who represent a population at risk for suicide. METHOD: We retrospectively reviewed charts regarding 783 out-patients consecutively admitted to a lithium clinic from 1976 to 1999. Individual age- and gender-specific quartile of serum cholesterol concentration were correlated against personal lifetime suicide attempts and completed suicide in first-degree relatives. RESULTS: The proportion of men with a personal lifetime history of attempted suicide, especially if violent, and that of men with history of completed suicide in a first-degree relative were significantly higher among the group with cholesterol concentration in the lowest quartile compared to the group with cholesterol levels above the 25th percentile. CONCLUSION: Low cholesterol concentration should be studied further as a potential biological/genetic marker of suicide risk. PMID- 11437749 TI - Relatives' distress and patients' symptoms and behaviours: a prospective study of patients with schizophrenia and their relatives. AB - OBJECTIVE: The aim of this study is to explore the relationship between relatives' distress and patients' symptoms and behaviours. METHOD: Fifty relatives in close contact with 36 patients with schizophrenia DSM-III-R filled in the General Health Questionnaire (GHQ) and the Perceived Family Burden Scale (PFBS) at the patient's hospital admission, 4.5 and 9 months post-discharge. The patients were assessed by means of the Positive and Negative Syndrome Scale (PANSS). RESULTS: The PFBS anxiety-depression cluster was at all three assessments positively correlated with relative's distress (GHQ), not with PANSS anxiety and depression measurements. In multiple regression analysis PFBS, but not PANSS, was related to relatives' distress. CONCLUSION: Relatives' distress was related to their reports of problematic patient behaviours, especially anxiety-depressive behaviour, not to symptoms as measured by clinical interviews. High distress is related to high expressed emotion, suggesting that relatives' report of patient's behaviour should be addressed to improve patient's outcome. PMID- 11437750 TI - Quality of life in schizophrenia in India: comparison of three approaches. AB - OBJECTIVE: To compare ratings of quality of life in patients with established schizophrenia in Chandigarh, India, in both subjective and objective view points and between patients' and relatives' ratings. METHOD: Subjective and objective quality of life was assessed in 38 patients with schizophrenia using the Quality of Life Interview - Brief Version and WHO Quality of Life - Bref scale. Key family members were also interviewed using the same scales. RESULTS: Subjective and objective quality of life are moderately well correlated (r = 0.43) and similar agreement was found between patients and relatives (r = 0.46). CONCLUSION: Although quality of life is characteristically measured with subjects, the use of objective measures of quality of life and key informants of patients provide useful proxy measures. PMID- 11437751 TI - Testing the stress-vulnerability hypothesis in ICD-10-diagnosed acute and transient psychotic disorders. AB - OBJECTIVE: To examine if family-history-positive (FHP) vis-a-vis family-history negative (FHN) probands with ICD-10-diagnosed acute and transient psychotic disorders (ATPD) differ significantly with respect to number of life events and cumulative stress score prior to the onset of ATPD. METHOD: Forty probands with ICD-10-based clinical diagnosis of acute and transient psychotic disorders were studied with respect to: (a) history of psychiatric disorders in first-degree relatives, and (b) antecedent life events and cumulative stress. The FHP and FHN probands were compared on the latter variables using Mann-Whitney U-test. RESULTS: FHP probands reported significantly less number of total life events compared to FHN probands (P = 0.006). Similarly, FHP probands had significantly less stress score than FHN probands (P = 0.002). There were no significant demographic or clinical differences between the two groups. CONCLUSION: The findings support the stress-vulnerability hypothesis in the aetiology of ATPD. PMID- 11437752 TI - Comparison of four groups of substance-abusing in-patients with different psychiatric comorbidity. AB - OBJECTIVE: Comparisons of different groups of dual patients are rare, yet potential differences could have therapeutic implications. In this study, four groups of psychiatric in-patients with substance use disorder were compared to each other: patients with no psychiatric comorbidity, patients with comorbid schizophrenia and patients with affective and personality disorder. METHOD: Apart from sociodemographic, therapy-related variables and a detailed survey of their substance use, all subjects were assessed with BPRS and SCL-90-R. RESULTS: No differences were found in the patients' demography, psychosocial adjustment and substance consumption career. Significant differences were found in regard to some therapy variables reflecting adherence to treatment and global outcome and to the level of psychopathology. CONCLUSION: Both substance use and comorbid psychiatric disorder have a variable impact on distinct areas of patients' general condition and functioning. The group with comorbid affective disorder appeared to be the most difficult to treat and the therapeutic approach to this disorder deserves reconsideration. PMID- 11437753 TI - Differences of symptoms and standardized weight index between patients with early onset and late-onset anorexia nervosa. AB - OBJECTIVE: There have so far been no studies that directly compared clinical features between patients with early- and late-onset anorexia nervosa (AN). METHOD: We identified 64 patients with DSM-III-R AN. We defined individuals as an early-onset group, who had an age of onset before 14 years (N = 31), and the remaining as a late-onset group (N = 33). The clinical symptoms, body weight and weight index, were compared between the two groups. Subjects were dichotomized into those with extremely low weight and those remaining. We compared the proportion of the patients with extremely low weight between the two groups. RESULTS: The rates of 'self-induced vomiting' and 'purging' were significantly lower in a group of patients with early-onset AN than in those with late-onset AN. There were significantly fewer subjects with extremely low weight in early onset than in late-onset AN group. CONCLUSION: We found clear differences in clinical features between early- and late-onset AN groups. PMID- 11437754 TI - Late-onset bipolar disorder due to hyperthyroidism. AB - OBJECTIVE: Bipolar disorder starts typically in early age and late-onset cases are rare. Late-onset cases are more likely to have comorbid medical illnesses responsible for them. This case report highlights late-onset bipolar disorder due to hyperthyroidism. METHOD: A 65-year-old patient of bipolar disorder has been described. RESULT: Physical examination and laboratory investigations detected presence of hyperthyroidism and the patient was treated with antithyroid and anxiolytics. CONCLUSION: A thorough examination and investigation are required in late-onset cases of bipolar disorder to rule out secondary causes. Definitive antimanic agents or mood stabilizers may not be required in such cases. PMID- 11437756 TI - Guidelines for lithium monitoring: are they ideal? PMID- 11437757 TI - Misdiagnosis of antidepressant discontinuation symptoms. PMID- 11437758 TI - When is a standard, not a standard? When it is a recommendation. PMID- 11437759 TI - Redistribution of halothane and sevoflurane under simulated conditions of acute airway obstruction. AB - Forty patients having surgery requiring muscle paralysis and tracheal intubation were randomly allocated to receive either halothane (n = 20) or sevoflurane (n = 20). Following intravenous anaesthesia and tracheal intubation, inhalation induction of anaesthesia was simulated. After attaining an end-tidal anaesthetic concentration of 2 MAC for the respective agent, the airway was obstructed for 3 min. The end-tidal anaesthetic concentration was measured for the first three breaths following the period of airway obstruction. The decrease in alveolar concentration of sevoflurane following 3 min of airway obstruction was found to be significantly greater than that of halothane. We conclude that even if the airway obstructs completely during inhalational induction of general anaesthesia, awakening would be faster with sevoflurane than with halothane. PMID- 11437760 TI - Combined pre-incisional oral dextromethorphan and epidural lidocaine for postoperative pain reduction and morphine sparing: a randomised double-blind study on day-surgery patients. AB - The reduction in acute pain perception following dextromethorphan has previously been investigated in patients undergoing general anaesthesia. This random and double-blind study examined the effects of pre-incisional oral dextromethorphan on postoperative pain and intravenous patient-controlled morphine demand in 60 day-surgery patients undergoing lower body surgery under lidocaine (1.6%-16 ml) epidural anaesthesia after receiving placebo, 60 or 90 mg dextromethorphan, 90 min pre-operatively. Postoperative pain was scored on a visual analogue scale from 1 to 10. In-hospital observation continued for 6 h and for 3 days at home; diclofenac was available throughout. Dextromethorphan-treated patients reported significantly (p < 0.05) less pain and sedation, and felt better. Patients who received dextromethorphan 90 mg had significantly (p < 0.05) lower heart and respiratory rates than those who received 60 mg. Medicated patients required half the morphine and diclofenac of placebo patients: 38% of patients who received 90 mg and 21% who received dextromethorphan 60 mg used no morphine or diclofenac whatsoever, a previously unreported finding. PMID- 11437761 TI - Transoesophageal acoustic quantification for evaluation of cardiac function during laparoscopic surgery. AB - Pneumoperitoneum may be poorly tolerated in patients with marginal cardiopulmonary function. Transoesophageal echocardiography is a relatively non invasive technique that can track changes in left ventricular cavity area using acoustic quantification and has been found to provide a reliable, real-time estimation of left ventricular function. We evaluated mid-papillary fractional area change during pneumoperitoneum in 17 healthy adult patients (group 1) and 17 with hypertension and/or ischaemic heart disease (group 2) undergoing laparoscopic cholecystectomy. Peritoneal insufflation led to significant (p < 0.01) decreases in fractional area change and an increase in mean blood pressure. There was no significant change in heart rate. Patients with cardiovascular disease who had a > 30% decrease in fractional area change following carbon dioxide insufflation (group 2B) were given an intravenous infusion of glyceryl trinitrate. In groups 1 and 2A, fractional area change improved slightly with head-up tilt and continued to improve over the course of surgery. Glyceryl trinitrate led to a significant increase in fractional area change and a decrease in mean blood pressure back to baseline values. We conclude that peritoneal insufflation of carbon dioxide results in a significant increase in cardiac workload that is well tolerated in patients with no cardiovascular disease. Coexisting cardiovascular disease can result in even more pronounced impairment of cardiac function, which may not be predictable by clinical assessment or adequately determined by routine monitoring. We recommend the use of transoesophageal acoustic quantification as a monitor in these patients and the administration of glyceryl trinitrate to ameliorate myocardial stress. PMID- 11437762 TI - Peri-operative silent myocardial ischaemia and long-term adverse outcomes in non cardiac surgical patients. AB - Two hundred and seventy-five non-cardiac surgical patients were recruited to determine risk factors associated with the development of postoperative cardiovascular complications during the first year after surgery. Patients underwent ambulatory electrocardiography pre- and postoperatively. There were 34 adverse events over the whole study period. Twenty-four occurred within 6 months and the remaining 10 occurred between 6 and 12 months postoperatively. Silent myocardial ischaemia was associated with adverse outcome over both the first 6 months [OR 4.44 (95% CI 1.77-11.13)] and the whole study period [OR 2.81 (1.26 6.07)]. Other risk factors were: vascular surgery [OR 17.09 (2.67-351.44)], history of angina [OR 6.29 (2.21-17.62)], concurrent treatment with calcium entry blockers [OR 2.68 (1.03-6.93)] and smoking [OR 4.93 (2.00-12.02)]. None of these was a useful predictor of long-term outcome (between 6 and 12 months postsurgery). These results are at variance with other published data, but we conclude that monitoring for peri-operative silent myocardial ischaemia does not aid the prediction of long-term cardiovascular complications. PMID- 11437763 TI - Fasting regimens for regional ophthalmic anaesthesia. A survey of members of the British Ophthalmic Anaesthesia Society. AB - Members of the British Ophthalmic Anaesthesia Society were surveyed using a postal questionnaire. The response rate was 72.3%. Respondents were asked about starvation before regional anaesthesia for cataract surgery, the use of sedation in these patients, monitoring and if oxygen supplementation was given. The results show that most patients are not starved before this type of regional anaesthesia, and that the majority of patients receive no supplementary sedation or intravenous analgesia. Over 70% of patients received oxygen supplementation. PMID- 11437764 TI - Descriptive patient data as an explanation for the variation in average daily costs in intensive care. AB - Intensive care patients require therapy that can vary considerably in type, duration and cost, so making it extremely difficult to predict patient resource use. Few studies measure actual costs; usually average daily costs are calculated and these do not reflect the variation in resource use between individual patients. The aim of this study was to analyse a data set of 193 critically ill adult patients to look for associations between routinely collected descriptive data and patient-specific costs. Regression analysis was used to explore any relationships between average daily patient-specific costs and the following variables: duration of intensive care unit stay, Acute Physiology and Chronic Health Evaluation II scores in the first 24 h, gender, age, mechanical ventilation at any point during the stay, postoperative status, emergency admission and mortality. Overall, this analysis explained 33.6% of the variation in average daily costs. The additional costs of an extra day of care, mechanical ventilation, an extra point on the Acute Physiology and Chronic Health Evaluation II score, and survival were obtained. PMID- 11437765 TI - Status epilepticus. AB - Status epilepticus is a medical emergency that requires rapid and vigorous treatment to prevent neuronal damage and systemic complications. Failure to diagnose and treat status epilepticus accurately and effectively results in significant morbidity and mortality. Cerebral metabolic decompensation occurs after approximately 30 min of uncontrolled convulsive activity, and the window for treatment is therefore limited. Therapy should proceed simultaneously on four fronts: termination of seizures; prevention of seizure recurrence once status is controlled; management of precipitating causes of status epilepticus; management of the complications. This article reviews current opinions about the classification, aetiology and pathophysiology of adult generalised convulsive status epilepticus and details practical management strategies for treatment of this life-threatening condition. PMID- 11437766 TI - An evaluation of the Airway Management Device. AB - We have evaluated the Airway Management Device (AMD) in 105 anaesthetised patients. We were successful in establishing a clear airway on the first attempt on 69 occasions and unable to establish a patent airway at all in 10 patients. Airway obstruction requiring removal of the device occurred during maintenance of anaesthesia in a further two cases and during emergence in three. Loss of the airway during anaesthesia occurred in eight of 95 patients and could be reversed by manipulation of the airway in six cases. Overall, a mean of 0.56 manipulations per patient were required to establish an airway and a further 0.42 per patient were required during maintenance of anaesthesia. In the 95 patients in whom an airway was established, assisted ventilation was satisfactory in 93, with a leak pressure above 20 cmH2O in 65. Intracuff pressure was measured in 12 cases and was above 100 cmH2O in eight. Minor complications occurred in 12 patients. Blood was visible on removal of the device in six cases. PMID- 11437767 TI - Anaesthetic management of 27 cases of boron neutron capture therapy for glioblastoma. AB - Twenty-seven patients received boron neutron capture therapy during craniotomy at our research reactor from 1991 to 1999. This is a form of intra-operative radiation therapy, which uses neutrons from a nuclear reactor. There are three additional major problems to anaesthetists: boron neutron capture therapy must be given beside the nuclear reactor, with no hospital facilities; neutrons cannot be shielded effectively by ordinary protectors; and neutrons are detrimental to metal devices and especially to electrical appliances. Boron neutron capture therapy has been adopted as an effective therapy for glioblastoma/astrocytoma, but special considerations are required for anaesthesia. PMID- 11437768 TI - Coronary artery bypass graft surgery in a patient with haemoglobin SC disease. AB - Patients with sickle cell disease who undergo surgery are generally considered to be at greater risk of peri-operative complications than otherwise healthy patients. We report a case of a woman with haemoglobin SC disease undergoing coronary artery bypass grafting. She was successfully managed with pre-operative exchange transfusion and normothermic cardiopulmonary bypass. PMID- 11437769 TI - A pilot study to investigate the effects of an infusion of aminophylline on renal function following major abdominal surgery. AB - Acute renal failure is a frequent complication of critical illness and optimal preventive therapy remains elusive. There is increasing evidence from animal models and some human studies that adenosine receptor antagonism by aminophylline may reduce the severity of renal impairment caused by a variety of aetiologies. We studied the renal effects of intravenous aminophylline in an unblinded, within patient study of 20 patients admitted to a general intensive care unit following major surgery. We demonstrated that there were no adverse cardiovascular complications related to aminophylline therapy. Renal sodium and osmolar clearance increased with a non-significant trend towards increased diuresis during treatment. Creatinine clearance, however, was unchanged but the study was not designed and did not have the power to test whether aminophylline increased renal blood flow or glomerular filtration rate. We suggest the renal actions of aminophylline in critical illness merit further investigation. PMID- 11437770 TI - Effects of surgical stress and nitrous oxide anaesthesia on peri-operative plasma levels of total homocysteine. A randomised, controlled study in general surgery. AB - Previous studies of patients have shown that anaesthesia with nitrous oxide (N2O) increases the plasma levels of total homocysteine. In a randomised, controlled trial we measured the plasma total homocysteine levels in patients undergoing general surgery before and after anaesthesia with and without N2O. Plasma total homocysteine levels were measured before anaesthesia and 1, 3-5 and 24 h after incision in 24 patients randomly allocated to anaesthesia with N2O (n = 12) and without N2O (n = 12). Total homocysteine levels significantly decreased from 10.4 +/- 2.7 to 8.2 +/- 2.9 micromol x l(-1) in the non-N2O group 24 h after incision (p < 0.02), while they tended to increase slightly in the N2O group from 10.5 +/- 4.5 to 10.9 +/- 4.3 micromol x l(-1) (p > 0.05). Our randomised controlled study indicates that total homocysteine decreases after general surgery in patients in whom anaesthesia is maintained without N2O, but not in patients in whom anaesthesia is maintained with N2O. PMID- 11437771 TI - Effectiveness of continuous positive airway pressure to enhance pre-oxygenation in morbidly obese women. AB - Morbid obesity is associated with a reduction in time to desaturate during apnoea following standard pre-oxygenation and induction of anaesthesia. We have compared the effects of using 7.5 cmH2O of continuous positive airway pressure (CPAP) for pre-oxygenation with a standard technique using a Mapleson A breathing system, in 20 morbidly obese women. In a prospective, open, randomised trial, we measured the time taken to desaturate to 90% from time of giving a succinylcholine bolus as part of a rapid induction of anaesthesia. All patients received 3 min pre oxygenation prior to induction. Tracheal intubation was confirmed and all patients kept apnoeic until oxygen saturation decreased to 90%. No statistically significant difference in mean time to desaturate to 90% could be demonstrated in the CPAP group compared to the Mapleson A group (240 s and 203 s, respectively). A brief period of lower mean heart rate in the CPAP group was the only statistically significant difference in cardiovascular parameters. There was no significant difference in the volume of gastric gas after induction between groups. PMID- 11437772 TI - Organisational failures in urgent and emergency surgery. A potential peri operative risk factor. AB - Medical error is an important cause of morbidity and mortality. Organisational failure in the pre-operative period has been associated with catastrophic outcome. Little information is available regarding peri-operative organisational problems. The incidence and nature of organisational failure before urgent and emergency surgery in a district general hospital was studied prospectively in 159 cases over a 30-day period. Organisational failure affected more than half of the cases overall, but varied in both its incidence and its complexity between surgical disciplines. Various causative factors were identified, e.g. 8% of cases were subject to delay due to clinical emergencies. The median [range] time required to rectify the problems was 115 [5-750] min. A consultant anaesthetist and surgeon were present in 30 and 20% of cases, respectively. Difficulty with the preparation of patients for emergency surgery is an important but underevaluated cause of medical error that may put patients at risk. PMID- 11437773 TI - Anaesthetic equipment: disposal or decontamination? PMID- 11437775 TI - A repainted nitrous oxide cylinder? PMID- 11437776 TI - Jugular bulb oximetry. PMID- 11437777 TI - Phaeochromocytoma: an unusual cause of hypertension in pregnancy. PMID- 11437778 TI - The value of case reports. PMID- 11437779 TI - Vagal nerve stimulators and anaesthesia. PMID- 11437780 TI - How invasive should one go? PMID- 11437781 TI - Preventing latex sensitisation and foreign body micro-emboli. PMID- 11437782 TI - Nitrous oxide. PMID- 11437783 TI - Post-operative pain relief using thoracic epidural analgesia. PMID- 11437784 TI - In favour of retaining anaesthetic induction rooms. PMID- 11437785 TI - Efficiency in intensive care--we must get it right. PMID- 11437786 TI - Fat embolism. PMID- 11437787 TI - The airway and the anaesthetist. PMID- 11437788 TI - Severe hypokalaemia causing acute respiratory failure. PMID- 11437789 TI - Nasogastric-tube-induced unilateral vocal cord palsy. PMID- 11437790 TI - A manoeuvre for using the flexible fibreoptic bronchoscope through the Intubating Laryngeal Mask Airway. PMID- 11437791 TI - Difficult airway--can intubate, can't ventilate. PMID- 11437792 TI - Challenges in paediatric mask holding; the 'claw hand' technique. PMID- 11437793 TI - Flexiblade laryngoscope. PMID- 11437794 TI - Another faulty catheter mount: now you see it now you don't. PMID- 11437795 TI - An unusual case of tracheal tube obstruction. PMID- 11437796 TI - Complete power failure. PMID- 11437797 TI - Child and adolescent psychiatry research comes of age. PMID- 11437798 TI - Challenges and benefits of assessment, diagnosis, and taxonomy for clinical practice and research. AB - OBJECTIVE: To outline nosological (top-down) and empirically based (bottom-up) approaches to assessment and taxonomy of psychopathology. METHOD: The two approaches were compared and contrasted to highlight similarities and differences between them. RESULTS AND CONCLUSIONS: To advance services and research, we need to make optimal use of assessment data (i) to identify the distinguishing features of each case, and (ii) to link individual patterns of functioning with taxonomic constructs that can help us apply previously accumulated knowledge to new cases. The time may be ripe for integrating nosological and empirically based approaches. One way to integrate these approaches is to construct empirically based and DSM-oriented scales from the same item pools and to generate age- and gender-specific standard scores and cut-off points from the same normative samples. Integration of the approaches was illustrated with profiles of empirically based and DSM-oriented scales scored from the same item pools and quantified in relation to the same normative samples. To facilitate use by clinicians and researchers under diverse conditions, data are readily obtained on forms completed independently by parents, teachers and others. The data can be quickly scored by hand or computer. To take account of situational and informant variations, the computer software systematically compares data from multiple informants on empirically based and DSM-oriented scales. PMID- 11437799 TI - Imaging normal and abnormal brain development: new perspectives for child psychiatry. AB - OBJECTIVE: The availability of non-invasive brain imaging permits the study of normal and abnormal brain development in childhood and adolescence. This paper summarizes current knowledge of brain abnormalities of two conditions, attention deficit hyperactivity disorder (ADHD) and childhood onset schizophrenia (COS), and illustrates how such findings are bringing clinical and preclinical perspectives closer together. METHOD: A selected review is presented of the pattern and temporal characteristics of anatomic brain magnetic resonance imaging (MRI) studies in ADHD and COS. These results are discussed in terms of candidate mechanisms suggested by studies in developmental neuroscience. RESULTS: There are consistent, diagnostically specific patterns of brain abnormality for ADHD and COS. Attention deficit hyperactivity disorder is characterized by a slightly smaller (4%) total brain volume (both white and grey matter), less-consistent abnormalities of the basal ganglia and a striking (15%) decrease in posterior inferior cerebellar vermal volume. These changes do not progress with age. In contrast, patients with COS have smaller brain volume due to a 10% decrease in cortical grey volume. Moreover, in COS there is a progressive loss of regional grey volume particularly in frontal and temporal regions during adolescence. CONCLUSIONS: In ADHD, the developmental pattern suggests an early non-progressive 'lesion' involving neurotrophic factors controlling overall brain growth and selected dopamine circuits. In contrast, in COS, which shows progressive grey matter loss, various candidate processes influencing later synaptic and dendritic pruning are suggested by human post-mortem and developmental animal studies. PMID- 11437800 TI - Global measures of impairment in children and adolescents: results from a Swiss community survey. AB - OBJECTIVE: Based on data from a Swiss epidemiological survey, both the parent and the child versions of two global measures of impairment, the Columbia Impairment Scale (CIS) and the Children's Global Assessment Scale (CGAS) were analysed with regard to validity and the effects of gender. METHOD: Concurrent and discriminant validity were evaluated by examining the associations with other indicators of impairment. Gender effects on impairment were considered in each type of analysis. RESULTS: Based on the total sample and on the girls' data, concurrent validity of the various impairment scores as computed by correlations with other indicators of psychological dysfunction was good. When based on the boys' data it was less convincing. Discriminant validity was estimated by comparing impairment scores between those using and not using services and was proven for all impairment scores. There were significant gender effects for almost all scores, indicating that girls in contrast to boys were more impaired and functioned less well when professional contacts and psychiatric diagnosis were considered. CONCLUSIONS: The two impairment measures are well suited to community studies. Gender and informant differences have to be considered when defining thresholds for caseness. The two scales could also be used clinically, for example for training and evaluation purposes and for the decision whether or not a child or adolescent requires treatment. PMID- 11437801 TI - The Christchurch Health and Development Study: review of findings on child and adolescent mental health. AB - OBJECTIVE: This paper provides an overview of the Christchurch Health and Development Study (CHDS) and a summary of findings relating to child and adolescent mental health. METHOD: The CHDS is a longitudinal study of a birth cohort of 1265 children born in the Christchurch (New Zealand) urban region during mid 1977. This cohort has now been studied from birth to age 21. RESULTS: The paper examines the ways in which the study has been able to examine a wide range of issues. Key issues examined include: (i) measurement of disorder (respondent effects; dimensionality; scales vs categories); (ii) prevalence and treatment of disorder; (iii) stability and continuity of disorders; (iv) the contribution of risk and aetiological factors (e.g. lead exposure, parental divorce, child abuse, family adversity, sexual orientation) to psychosocial adjustment; and (v) the psychosocial consequences of mental health problems in adolescence. CONCLUSIONS: The study findings illustrate the many advantages of a longitudinal study, such as the CHDS, in providing methodologically sound, theoretically relevant and cost effective research that caters for the interests of multiple end-users including the scientific community, clinicians and applied policy makers. PMID- 11437802 TI - Longitudinal predictors of behavioural adjustment in pre-adolescent children. AB - OBJECTIVES: Longitudinal data from infancy onwards, from the Australian Temperament Project, a prospective study of the temperament and development of a large and representative sample of Victorian children, were examined to identify predictors of psychological disorder at 11-12 years of age. METHOD: Those children scoring in the at-risk range for psychological disorder according to parents, teachers and self-reports using the Child Behaviour Questionnaire were selected at 11-12 years of age for in-depth assessment and comparison with a group of children with no history of adjustment problems. Analyses of group differences using longitudinal data gathered from infancy to 12 years focused on parent and teacher reports on child temperament and behaviour, and various facets of home and school adjustment. RESULTS: The strongest predictors of adjustment at 12 years were previous behaviour problems, along with some specific temperament factors involving self-regulation capacities and mother's overall rating of child difficulty. Results based on parallel teacher data including peer adjustment, and social and academic competence measures were consistent with parent data. CONCLUSIONS: Our research confirms the persistence of early appearing behaviour problems in a community sample and the longitudinal influence of temperament factors in childhood. The study supports the need for a focus on early intervention and prevention strategies in the child mental health field. PMID- 11437803 TI - Child and adolescent psychiatry in general practice. AB - OBJECTIVE: This review discusses the role of general practitioners and primary care health staff in the assessment, treatment and prevention of child and adolescent psychiatric disorders and to consider implications for service delivery. METHOD: Literature review. Papers were selected as having an empirical evidence base or as describing and documenting new initiatives in general practice. RESULTS: A small proportion of about 3% of children present to general practice with behavioural or emotional problems, but psychiatric disorders in the context of somatic presentations are considerably more common. They are probably higher than in the general population reflecting an increased tendency by children with disorders to consult. Recognition by general practitioners is limited and few children with disorders are referred to specialist clinics. However, referrals are mostly appropriate: the more severely affected children in difficult psychosocial circumstances. A number of pilot studies have shown the feasibility and potential usefulness of setting up shifted specialist clinics in primary care and of training primary care doctors and other staff in the recognition or management of child mental heath problems. CONCLUSION: Primary care is an appropriate resource to help increase attention to child and adolescent mental health problems. Its potential requires further development and rigorous evaluation. Areas lending themselves to development include: improved medical undergraduate teaching and postgraduate training; suitable information and advice-giving on child mental health problems by the primary care team; the development of specific child and adolescent psychiatric interventions for use in the primary care setting; careful and discriminating development of shifted outpatient clinics for selected child psychiatric disorders; the development of focused protocols for referral to specialist services; further development of mental health promotion clinics in primary care. PMID- 11437804 TI - 'Why getting fat, Doc?' Weight gain and psychotropic medications. AB - OBJECTIVES: Weight gain associated with the use of psychotropic medications is a common clinical problem that is of particular importance because of its effects on the general health of psychiatric patients and their compliance with treatment. This paper aims to explore this issue and discuss the mechanisms of weight gain and methods of prevention. METHOD: A literature review (Index Medicus/Medline) was carried out as well as a review of other relevant papers and data known to the authors. RESULTS: Significant weight gain may result in considerable morbidity. The majority of psychotropic medications are associated with weight gain, however, the mechanisms of weight gain are often complex and poorly understood. CONCLUSION: Clinically, weight gain can be anticipated and often managed with some success in the majority of psychiatric patients with simple but relatively effective measures. It is important for clinicians to be aware of this common clinical problem and educate patients from the outset, monitoring them regularly and intervening when necessary. PMID- 11437805 TI - Major depression: does a gender-based down-rating of suicide risk challenge its diagnostic validity? AB - OBJECTIVE: This paper will summarize the authors' research that disproved the accepted lifetime suicide risk in major depression. It will then explore the pivotal issue of gender in understanding suicide risk in depression and raise questions as to whether this is adequately reflected in the current diagnostic construct of this condition. METHOD: The methods of two recent papers published by the authors are briefly recounted. In the first of these papers, an age specific algorithm was developed to reflect the necessary mathematical relationship between the prevalence of major depression, total population suicide rates and suicide risk in depression. It allowed for deaths in each age group from other causes, corrected for official underreporting, and was calculated on the entire population of the USA. In the second paper this methodology was further refined and applied to gender and age data. RESULTS: The suicide risk in major depression as it is currently defined diagnostically is of the order of 3.4% rather than the previously accepted figure of 15%. However, a single figure is misleading as it averages two highly disparate figures of almost 7% for men and only 1% for women. In youths (< age 25) the male: female ratio is even higher (10:1). CONCLUSIONS: Among sufferers of major depression, men and those who have been hospitalized have a much greater risk of suicide. These findings are sensitive to diagnostic inclusivity (the algorithm's denominator) which raises the question as to whether women with a depressive illness are more likely to be correctly identified than male sufferers? An argument is made for a gender-based nosological revision of the diagnostic criteria. In the interim, given the treatable morbidity of depression and the availability of safe, well-tolerated antidepressants, there is a prima facie case for lowering our threshold of treatment in men and youths presenting with a history of anger dyscontrol, or substance abuse, who have decompensated from previous levels of functioning and who show features of either typical or atypical depression. PMID- 11437806 TI - Depression and smoking: examining correlates in a subset of depressed patients. AB - OBJECTIVE: The objective of this study was to examine for associations between depression and cigarette smoking. METHOD: A sample of 92 depressed smokers was compared with a control sample of depressed non-smokers, matched for age, gender and diagnostic variables. Comparisons were made across a range of demographic, depression, family history, developmental factors, anxiety and personality style variables, as well as use of alcohol and illicit drugs. RESULTS: We failed to find any difference between smokers and non-smokers in history or severity of depression. Cigarette smokers were distinguished principally by greater exposure to aversive experiences in childhood, disordered personality function, greater use of illicit drugs, anxiolytics and alcohol. Logistic regression identified dysfunctional personality 'domains', physical violence in childhood, long-term anxiolytic use and illicit drug use as the most significant predictor set. CONCLUSIONS: Results favour a model of cigarette smoking and depression as linked by shared early deprivational variables, rather than cigarette smoking causing depression or the converse. PMID- 11437807 TI - Depressive disorders in elderly Chinese patients in Macau: a comparison of general practitioners' consultations with a depression screening scale. AB - OBJECTIVE: The objectives of this study were to identify the prevalence of depressive symptomatology in an elderly population using a screening scale; to assess general practitioner (GP) recognition of depression; to determine patient reasons for consultation; and to relate depression to social and demographic factors. METHOD: An analytical cross-sectional study was carried out on a systematic sample of elderly Chinese patients currently resident in Macau, who presented to general practices in December 1997. Data collected included patient demographics, economic and social supports, patient depressive status assessed using the Hospital Anxiety and Depression scale (HAD), reason for attendance, and the reason for the consultation. Clinical records were analysed in order to collect GP-registered morbidity and recent relevant management. RESULTS: Data were collected from 386 elderly Chinese patients of 31 GPs (97% patient response rate). Patients presented with somatic symptoms rather than psychological issues. Using HAD cut-off score of >8, 47% of the population studied were depressed; with a HAD subscale score >11, 26.2% were depressed. Depression was detected more often in women (59%) and being female was associated with depressive status (p = 0.010). Age (greater than 75 years) was also associated with being depressed, as was not having someone to talk to (p = 0.037), and being from low social class (p = 0.050). Not having someone willing to listen to their problems, difficulties and worries, was significantly related to depression (p = 0.041). There were no referrals or antidepressive medications detected in the clinical charts. CONCLUSION: Participants' characteristics such as being over 75 years of age, being female and socially isolated were related to depression. General practitioners can play an important role in the early detection and management of psychological disorders. Although studies on a sample of Chinese patients in Macau cannot be generalized to Chinese populations elsewhere, awareness of somatic presentation of psychological illness is crucial in detecting depression in this and possibly other Chinese populations. PMID- 11437808 TI - The adjustment of children of Australian Vietnam veterans: is there evidence for the transgenerational transmission of the effects of war-related trauma? AB - OBJECTIVE: The presence of posttraumatic stress disorder (PTSD) in trauma survivors has been linked with family dysfunction and symptoms in their children, including lower self-esteem, higher disorder rates and symptoms resembling those of the traumatized parent. This study aims to examine the phenomenon of intergenerational transfer of PTSD in an Australian context. METHOD: 50 children (aged 16-30) of 50 male Vietnam veterans, subgrouped according to their fathers' PTSD status, were compared with an age-matched group of 33 civilian peers. Participants completed questionnaires with measures of self-esteem, PTSD symptomatology and family functioning. RESULTS: Contrary to expectations, no significant differences were found between the self-esteem and PTSD symptomatology scores for any offspring groups. Unhealthy family functioning is the area in which the effect of the veteran's PTSD appears to manifest itself, particularly the inability of the family both to experience appropriate emotional responses and to solve problems effectively within and outside the family unit. CONCLUSION: Methodological refinements and further focus on the role of wives/mothers in buffering the impact of veterans' PTSD symptomatology on their children are indicated. Further effort to support families of Veterans with PTSD is also indicated. PMID- 11437809 TI - Prevalence of psychological distress and psychiatric disorders among homeless youth in Australia: a comparative review. AB - OBJECTIVE: To review the prevalence literature on psychological distress and psychiatric disorders among homeless youth in Australia, and to compare these rates with Australian youth as a whole. METHOD: Computerized databases were utilized to access all published Australian studies on psychological distress (as measured by standardized symptom scales and suicidal behaviour) and psychiatric disorders among homeless youth; in addition, unpublished Australian studies were utilized whenever accessible. A total of 14 separate studies were located, only three of which have included non-homeless control groups. In the current review, prevalence data from uncontrolled youth homelessness studies are compared with data from Australian community and student surveys. RESULTS: Homeless youth have usually scored significantly higher on standardized measures of psychological distress than all domiciled control groups. Youth homelessness studies have also reported very high rates of suicidal behaviour, but methodological limitations in these studies make comparisons with community surveys difficult. Furthermore, rates of various psychiatric disorders are usually at least twice as high among homeless youth than among youth from community surveys. CONCLUSIONS: Homeless youth in Australia have extremely high rates of psychological distress and psychiatric disorders. As homeless youth are at risk of developing psychiatric disorders and possibly self-injurious behaviour the longer they are homeless, early intervention in relevant health facilities is required. PMID- 11437810 TI - SPECT scan in somatization disorder patients: an exploratory study of eleven cases. AB - OBJECTIVE: There are no previous studies using single photon emission computed tomography (SPECT) scans in somatization disorder (SD) patients. The aim of this paper is to assess SPECT imaging abnormalities in SD patients and study any relation to laterality. METHOD: Eleven SD patients from the Somatization Disorder Unit of Miguel Servet University Hospital, Zaragoza, Spain, not fulfilling criteria for any other psychiatric disorder and showing normal computed tomography (CT) and magnetic resonance imaging (MRI) images were studied with SPECT. Patients with DSM-IV axis I comorbidity were ruled out because it has been demonstrated that SPECT scans can show abnormalities in patients with depression and anxiety disorders. The technique used for SPECT was 99mTc D,1,hexamethylpropyleneamide-oxime (99mTc-HMPAO) in four patients and 99mTc bicisate in the other seven. The SPECT scans were evaluated without knowledge of clinical data and entirely by visual inspection. RESULTS: Seven out of 11 (63.6%) SD patients showed hypoperfusion in SPECT imaging. In four cases there was hypoperfusion in the non-dominant hemisphere and the predominance of pain symptoms took place in the contralateral hemibody. In the other three patients hypoperfusion was bilateral. The anatomical regions affected were cerebellum (four cases), frontal and prefrontal areas (three cases), temporoparietal areas (two cases) and the complete hemisphere (one case). CONCLUSIONS: A proportion of SD patients may present hypoperfusion in SPECT images, uni- or bilaterally, in different brain areas. Possible aetiological explanations for this finding are discussed. Controlled studies are necessary to confirm or refute this hypothesis. PMID- 11437811 TI - Inquiries into homicides and serious violence perpetrated by psychiatric patients in New Zealand: need for consistency of method and result analysis. AB - BACKGROUND: The violent action of mentally ill people is a source of considerable public and professional concern. At times such incidents are subject to inquiry. In England and Wales, homicides by people suffering from mental illness are subject to mandatory external inquiry. Further, the Royal College of Psychiatrists coordinates a confidential research study into homicide and suicide by people in contact with mental health services. Inquiries have raised concern regarding widespread problems in mental health service delivery. Within New Zealand, similar concerns have been raised, but inquiries have been irregular and not of consistent methodology. The paper aims to review 10 years of inquiries into violent incidents to describe their methods, structure and findings. METHOD: All inquiries into violent actions perpetrated by patients in contact with mental health services between 1988 and 1998 and held by the Ministry of Health were reviewed. The nature of the inquiry, the incident, findings and recommendations were summarized. For each inquiry, an assessment was made as to whether the incident was predictable or preventable. RESULTS: There were 11 incidents leading to 13 inquiries, six of homicide, two of rape, one of the release of a dangerous patient and two in which a patient was shot by police. Two internal inquiries were followed by external inquiries. All inquiries found deficiencies of varying severity, the degree of deficiency being greater with external inquiries. Consistent criticisms related to skill, resource, coordination and communication failures. Two of the 11 inquiries found the incident to be 'predictable', and eight to have been 'preventable'. CONCLUSIONS: The problems in New Zealand are similar to those noted in England and Wales. Small numbers of inquiries make firm conclusions difficult, but the authors feel that a mandatory process of independent review of serious incidents is wise. PMID- 11437812 TI - Routine outcome measurement in a child and adolescent mental health service: an evaluation of HoNOSCA. The Health of the Nation Outcome Scales for Children and Adolescents. AB - OBJECTIVE: This paper evaluates a range of properties for a clinician-based instrument designed for routine use in a child and adolescent mental health service (CAMHS). METHOD: The Health of the Nation Outcome Scales for Children and Adolescents (HoNOSCA) is a new outcome measure with great promise. Case vignettes were used to examine interrater reliability. HoNOSCA was implemented for routine outpatient use by multidisciplinary staff with a return rate of 84%. The 305 ratings obtained at assessment were analysed by age, gender and diagnosis. A sample of 145 paired ratings with a 3-month interval were examined for the measurement of change over time. RESULTS: Interrater reliability of the total score indicates moderate reliability if absolute scores are used and good reliability if the total score is used for relative comparisons. Most scales have good to very good reliability. The scales discriminated between age and gender in the expected way. HoNOSCA correlated with clinicians' views of change and was sensitive to change over a 3-month period. The total score seemed a proxy for severity. CONCLUSION: Routine outcome instruments must be explored in settings where they will be used and with realistic training. HoNOSCA appears to be of value in routine outcome measurement and although questions remain about reliability and validity, the results strongly support further investigation. PMID- 11437813 TI - Measuring mental health outcomes in a private psychiatric clinic: Health of the Nation Outcome Scales and Medical Outcomes Short Form SF-36. AB - OBJECTIVE: This study reports on data collected from the routine use of the Health of the Nation Outcome Scales (HoNOS) and the Medical Outcomes Short Form (SF-36). Three main aims were addressed in using these measures: (i) to establish patient disability levels; (ii) to determine the level of treatment effectiveness; and (iii) to explore the ability of these instruments to predict length of stay and mood change. METHOD: The clinician-rate HoNOS and the patient rated SF-36 were included in the assessment battery, at admission and discharge, of consecutive inpatients (n = 754) at one private psychiatric facility over a 2 year period. RESULTS: The sample, on admission, was comparable in illness severity to levels reported at other Australian private psychiatric facilities. Treatment was shown to be effective, and the degree of changes in HoNOS ratings compared favourably with other private psychiatric facilities. Certain factors underlying the structure of the HoNOS and the SF-36 only weakly predicted length of stay and changes in depression and anxiety levels. CONCLUSION: The HoNOS and the SF-36 provided valid and reliable data on patient function, with the HoNOS being most sensitive to treatment change. However, neither instrument proved useful in predicting length of stay or levels of depression and anxiety at discharge. PMID- 11437814 TI - Asystole during electroconvulsive therapy: a case report. AB - OBJECTIVE: The objective of the report is to describe a case of asystole lasting for 18 s, which developed after a subconvulsive stimulus during electroconvulsive therapy (ECT) in a patient without pre-existing cardiovascular abnormality. A brief review of the relevant literature is also provided. CLINICAL PICTURE: The patient was a 65-year-old Chinese man with a 2-year history of depression and good past medical health. Earlier he had responded well to a course of ECT without adverse effects. This time he presented with low mood, anhedonia, poor appetite and constipation. He did not respond to adequate trials with several antidepressant medications. When a subconvulsive stimulus was administered to determine the seizure threshold, no seizure activity was detected. However, immediately after the stimulus the patient developed an 18-s asystole, followed by bradycardia of 40 beats per minute for 10 s. The bradycardia resolved spontaneously before therapeutic intervention was effected. TREATMENT: Intravenous atropine was employed as premedication and suprathreshold stimulus was used in further ECT sessions. OUTCOME: Asystole did not recur in the subsequent six ECT sessions. CONCLUSION: When proper precautions are taken, asystole does not necessitate the suspension of further ECT sessions but intravenous atropine should be considered as premedication in such cases. Inducing anaesthesia with methohexital, avoiding excessive amounts of succinylcholine and employing suprathreshold stimulus and unilateral electrode placement may further lessen the likelihood of asystole in susceptible cases. PMID- 11437815 TI - Can seizures and rhabdomyolysis be a potentially serious complication of hyponatremia due to polydipsia? PMID- 11437816 TI - Response to 'signal transmission, rather than reception, is the underlying neurochemical abnormality in schizophrenia'. PMID- 11437818 TI - Can we justifiably draw epidemiological conclusions from small case studies? PMID- 11437820 TI - Welcome managed care. PMID- 11437821 TI - Our journal's wrapping. PMID- 11437822 TI - Mood stabilizers in Asperger's syndrome. PMID- 11437823 TI - Acute severe catatonia in a young woman with chronic schizophrenia responding to parenteral clonazepam. PMID- 11437824 TI - Suicide and attempted suicide in the army in Italy 1986 to 1998. PMID- 11437825 TI - Response to 'better mental health services for young people: responsibility, partnerships and projects'. PMID- 11437826 TI - Response to 'traumatic stress in the 21st century'. PMID- 11437830 TI - Leishmania donovani lipophosphoglycan causes periphagosomal actin accumulation: correlation with impaired translocation of PKCalpha and defective phagosome maturation. AB - Lipophosphoglycan (LPG) is the major surface glycoconjugate of Leishmania donovani promastigotes. The repeating disaccharide-phosphate units of LPG are crucial for promastigote survival inside macrophages and establishment of infection. LPG has a number of effects on the host cell, including inhibition of PKC activity, inhibition of nitric oxide production and altered expression of cytokines. LPG also inhibits phagosomal maturation, a process requiring depolymerization of periphagosomal F-actin. In the present study, we have characterized the dynamics of F-actin during the phagocytosis of L. donovani promastigotes in J774 macrophages. We observed that F-actin accumulated progressively around phagosomes containing wild-type L. donovani promastigotes during the first hour of phagocytosis. Using LPG-defective mutants and yeast particles coated with purified LPG, we obtained evidence that this effect could be attributed to the repeating units of LPG. LPG also disturbed cortical actin turnover during phagocytosis. The LPG-dependent accumulation of periphagosomal F actin correlated with an impaired recruitment of the lysosomal marker LAMP1 and PKCalpha to the phagosome. Accumulation of periphagosomal F-actin during phagocytosis of L. donovani promastigotes may contribute to the inhibition of phagosomal maturation by physically preventing vesicular trafficking to and from the phagosome. PMID- 11437831 TI - Transcripts of the malaria vector Anopheles gambiae that are differentially regulated in the midgut upon exposure to invasive stages of Plasmodium falciparum. AB - Understanding the interactions between the most deadly malaria parasite, Plasmodium falciparum, and its main vector, Anopheles gambiae, would be of great help in developing new malaria control strategies. The malaria parasite undergoes several developmental transitions in the mosquito midgut and suffers population losses to which mosquito factors presumably contribute. To identify such factors, we analysed An. gambiae midgut transcripts whose expression is regulated upon ingestion of invasive or non-invasive forms of P. falciparum using a differential display approach. Sixteen cDNA were studied in detail; 12 represent novel genes of An. gambiae including a gene encoding profilin. Four transcripts were specifically regulated by P. falciparum gametocytes (invasive forms), whereas the others were regulated by either non-invasive or both non-invasive and invasive forms of the parasite. This differential regulation of some genes may reflect the adaptation of P. falciparum to its natural vector. These genes may be involved in the development of P. falciparum in An. gambiae or in the defence reaction of the mosquito midgut towards the parasite. PMID- 11437832 TI - Reversible interruption of Giardia lamblia cyst wall protein transport in a novel regulated secretory pathway. AB - To survive in the environment and infect a new host, Giardia lamblia secretes an extracellular cyst wall using a poorly understood pathway. The two cyst wall proteins (CWPs) form disulphide-bonded heterodimers and are exported via novel encystation-specific secretory vesicles (ESVs). Exposure of eukaryotic cells to dithiothreitol (DTT) blocks the formation of disulphide bonds in nascent proteins that accumulate in the endoplasmic reticulum (ER) and induces an unfolded protein response (UPR). Proteins that have exited the ER are not susceptible. Exposure to DTT inhibits ESV formation by > 85%. Addition of DTT to encysting cells causes rapid (t1/2 < 10 min), reversible disappearance of ESVs, correlated with reduction of CWPs to monomers and reformation of CWP oligomers upon removal of DTT. Neither CWPs nor ESVs are affected by mercaptoethanesulphonic acid, a strong reducing agent that does not penetrate cells. DTT does not inhibit the overall protein secretory pathway, and recovery does not require new protein synthesis. We found evidence of protein disulphide isomerases in the ESV and the surface of encysting cells, in which they may catalyse initial CWP folding and recovery from DTT. This is the first suggestion of non-CWP proteins in ESVs and of enzymes on the giardial surface. DTT treatment did not stimulate a UPR, suggesting that Giardia may have diverged before the advent of this conserved form of ER quality control. PMID- 11437833 TI - Expression of constitutively active Rab5 uncouples maturation of the Salmonella containing vacuole from intracellular replication. AB - The enteric bacterial pathogen Salmonella typhimurium enters and proliferates within both phagocytic and non-phagocytic host cells. Upon entry, the bacteria reside in membrane-bound vacuoles (SCVs) that mature with time, as evidenced by the sequential loss of early endosomal markers, followed by the selective recruitment of a number of lysosomal membrane glycoproteins (LAMPs). This remodelling process renders the SCVs non-fusogenic with lysosomes and is also thought to create a vacuolar environment permissive for replication. We demonstrate that disruption of the endocytic pathway by the expression of a constitutively active form of the small GTPase rab5 (rab5Q79L) significantly altered the biogenesis of the SCVs without inhibiting bacterial replication in HeLa cells. Expression of rab5Q79L caused the retention of early endosomal markers on SCVs and early acquisition of LAMP2, and led to an increase in the kinetics of intracellular replication. We also demonstrate that a significant fraction of LAMP2 in SCVs is derived from the cell surface via endocytosis rather than via the biosynthetic route. Further, in fibroblasts lacking a functional AP3 adaptor complex, in which all newly synthesized LAMP is delivered to the cell surface, recruitment of LAMP to the SCVs remained unaffected. These findings raise the possibility that all the SCV-associated LAMP could be derived by endocytosis from the cell surface. PMID- 11437834 TI - Identification of Brucella spp. genes involved in intracellular trafficking. AB - After uptake by host cells, the pathogen Brucella transits through early endosomes, evades phago-lysosome fusion and replicates in a compartment associated with the endoplasmic reticulum (ER). The molecular mechanisms underlying these processes are still poorly understood. To identify new bacterial factors involved in these processes, a library of 1800 Brucella melitensis 16M mini-Tn5catkm mutants was screened for intracellular survival and multiplication in HeLa cells and J774A.1 macrophages. Thirteen mutants were identified as defective for their intracellular survival in both cell types. In 12 of them, the transposon had inserted in the virB operon, which encodes a type IV-related secretion system. The preponderance of virB mutants demonstrates the importance of this secretion apparatus in the intracellular multiplication of B. melitensis. We also examined the intracellular fate of three virB mutants (virB2, virB4 and virB9) in HeLa cells by immunofluorescence. The three VirB proteins are not necessary for penetration and the inhibition of phago-lysosomal fusion within non professional phagocytes. Rather, the virB mutants are unable to reach the replicative niche and reside in a membrane-bound vacuole expressing the late endosomal marker, LAMP1, and the sec61beta protein from the ER membrane, proteins that are present in autophagic vesicles originating from the ER. PMID- 11437835 TI - Acknowledging the extra care parents give their disabled children. AB - Around 150,000 families in the UK care for a severely disabled child under the age of 16. Many of these families receive assistance from the Family Fund Trust, which provides grants and information relating to the care of a severely disabled child. The aim of this study was to identify patterns of extra care needs among severely disabled children known to the Trust. Extra care needs are requirements for care not experienced by similarly aged non-disabled children. The research comprised analysis of 40,000 records from the Trust database and qualitative exploration of the extra care needs of disabled children with parents and Trust staff. Although all children require parenting, the care parents give disabled children generally exceeds that given to a non-disabled child. Quantitative analysis showed that the majority of children in the sample required extra assistance or supervision with multiple areas of daily life. With each of five activities (washing, dressing, meal times, during the night and keeping occupied), >70% of children needed extra help and, on average, each child needed extra help or supervision in six areas of daily life. Cluster analysis indicated distinctive combinations of extra care needs. Qualitative material indicated variety in extra care tasks undertaken (physical help, supervision, guidance) and causal factors (physical limitations, cognitive difficulties, behavioural problems). The findings confirm that severely disabled children have considerable extra care needs in many areas of daily life. Parents want professionals to recognize and offer explicit acknowledgement of the extra care they give their disabled children. PMID- 11437836 TI - Technology-dependent children in the community: definitions, numbers and costs. AB - OBJECTIVES: To assess available data on the numbers of technology-dependent children living at home in the UK and estimate the costs of caring for them. DESIGN: Data were obtained from all known secondary sources, including surveys of children with specific conditions known to specialist paediatric departments and the database of applicants to the Family Fund Trust. The costs of all services received by three hypothetical exemplar 'case study' children were calculated. RESULTS: All existing sources of data have some shortcomings. Taking these shortcomings into account, it is estimated that there may be up to 6000 technology-dependent children living at home. They appear to be disproportionately young and may be distributed unevenly between different regions of the UK. The total service costs of caring for each child will vary according to the types of technologies involved and local patterns of services, but may be as high as 150,000 pounds sterling a year. CONCLUSIONS: Because of the very high costs of services, routine information on the numbers of technology dependent children discharged home is urgently needed. This should include details of the duration of technology dependence and the local area to which the child is discharged. New opportunities for joint service planning and purchasing should improve the co-ordination of services for these children. PMID- 11437837 TI - Covert fears and anxiety in asthma and congenital heart disease. AB - AIM: To compare anxiety, fears and behavioural problems in children with asthma and children with congenital heart disease, and with the normative population. To also review the influence of maternal anxiety, time since diagnosis and severity of disease. DESIGN: Children administered Fear Survey Scale (FSSC-R) and Child Manifest Anxiety Scale (R-CMAS). Mothers given Child Behaviour Checklist (CBCL) and State Trait Anxiety Scale (STAI-S and STAI-T). Normative means and SDs compared with means and SDs for both medical groups. The mother's scores on the STAI-S and STAI-T scales were correlated with the child's scores on the FSSC-R and the R-CMAS. SETTING: Outpatient Asthma and Cardiology multidisciplinary Clinics at a tertiary care paediatric facility, Alberta Children's Hospital. SUBJECTS: Forty children with asthma (aged 6--17 years) were compared with 39 children with congenital heart disease. Intake questionnaires and interviews determined these children to be without obvious psycho-social problems. RESULTS: Children with asthma and children with congenital heart disease had more medical fears, and more physiological anxiety than normative samples. Increased maternal anxiety was correlated in both groups with increased child anxiety, medical fears and behavioural problems in the child. Similarly, increased severity of asthma or cardiac problems was associated with more physiological anxiety and more fears. Less time since diagnosis of the disease adversely affected social interactions in both groups of children. CONCLUSION: Physiological anxiety, medical fears and maternal anxiety are important issues requiring attention in asthma and cardiac disease, even in the absence of obvious psychosocial problems. There may be specific problems with a recent diagnosis of a chronic illness. PMID- 11437838 TI - Treatment demands and differential treatment of patients with cystic fibrosis and their siblings: patient, parent and sibling accounts. AB - Cystic fibrosis (CF) is a progressive disease with no known cure. Advances in diagnosis and treatment have resulted in patients living longer and thus families live with the illness for longer. Treatments are becoming increasingly demanding and are largely performed in the family home. Mothers are often reported to experience greater stress and poorer adjustment than mothers of well children or population norms. Patients and siblings are also reported to display adjustment difficulties. Siblings have rarely been included in research designs. This qualitative study investigates the impact of CF and treatment on eight patients, eight mothers, one father and eight siblings. A family systems perspective was adopted. Each individual was interviewed independently using semistructured interviews. Patients and siblings were aged between 9 and 21 years. Qualitative analyses revealed high levels of non-adherence (intentional and unintentional) and parental involvement in treatment, minimal involvement of siblings, and preferential treatment towards patients. Demanding treatment, coupled with the progressive nature of CF, promote high levels of parental involvement for younger children as well as older teenagers, often due to attempted or actual non adherence. Siblings may receive less attention while patients' needs take priority. Future development of a measure of adherence suitable for children and adolescents should take into account different motivations for non-adherence, particularly regarding the level of personal control over adherence to treatment. In addition, the potential impact of having a brother or sister with CF should not be underestimated and the needs of siblings should not go unnoticed. PMID- 11437839 TI - Measuring everyday visual discrimination in visually impaired children with the Sonksen Picture Guide to Visual Function. AB - The Sonksen Picture Guide to Visual Function (SPGVF) assesses a person's ability to discriminate pictures of everyday objects. Sonksen added the SPGVF to tests of visual acuity, because real objects, miniatures, and pictures are the main learning medium for young children. In earlier studies correlations have been found between visual acuity and responses to the SPGVF. However, to date no studies have used visually impaired children. In the study reported here, the associations between performance on the SPGVF task, visual acuity and contrast sensitivity were investigated. Participants were 17 visually impaired and 29 children without disabilities, 4--6 years of age. The results showed that visual acuity was more important for picture recognition than contrast sensitivity. Contrast thresholds < 3.33% implied that not all SPGVF-pictures could be recognized. The visually impaired children in the current study showed shorter mean identification distances than the normally sighted children in the original Sonksen and Macrae study. PMID- 11437841 TI - Investigation for immunodeficiency in patients with recurrent ENT infections. AB - Patients suffering with primary immunodeficiency frequently present to ear, nose, and throat (ENT) clinics, but the diagnosis is rarely made at this time. Early diagnosis of these patients would help to prevent morbidity and even mortality. Normal results from a simple panel of blood tests will exclude the commonest immune deficiencies. An abnormal result from these tests, or a strong suspicion despite normal initial testing, should prompt discussion with an immunologist. PMID- 11437842 TI - A prospective case-control study of 50 consecutive patients presenting with hyperlipidaemia. AB - The literature contains many references which refer to a causal relationship between hyperlipidaemia and hearing loss, but the majority of reports lack adequate controls, or are based on a series of cases which may represent incidental findings. This prospective case-control study compared a restricted population of 50 consecutive hyperlipidaemic patients attending a lipid clinic whose fasting lipid levels were > 2 SDs above the population mean with a control population recruited from patients undergoing nasal surgery for structural abnormalities (n = 159). The National Study of Hearing data was also compared with both groups in order to provide external validity to the control group. This study showed no consistent differences in the hearing thresholds of the hyperlipidaemic group compared with either control group. The Null hypothesis that sensorineural hearing loss is no greater in a population whose fasting blood lipids are raised > 2 SDs above the population mean level than in a control population cannot be refuted given the variability of the data. PMID- 11437843 TI - Waiting for ENT opinion and treatment: will things ever improve? PMID- 11437844 TI - Augmentation of the nasal dorsum using Gore-Tex: intermediate results of a retrospective analysis of experience in 66 patients. AB - Out of the numerous implant materials that have been used for augmentation of the nasal dorsum, autogenous cartilage is considered to be the optimal grafting material due to its versatility and long-term survival. However, in the case of extensive augmentation often an alternative grafting material is needed. Homologous cartilage seems an attractive option, but is not commonly used because of the fear of disease transmission, the long-term unpredictability and the possibility of warping. Alloplastic grafts have offered varying degrees of success in rhinoplasty, but have resulted in significant complications as well. Lately, expanded polytetrafluorethylene (Gore-Tex(R)) has proven to be a promising synthetic material in nasal dorsal augmentation. However, convincing long-term success compared to autogenous cartilage grafts is still lacking and numbers are insufficient. This paper consists of a prelimary report about the use of Gore-Tex(R) soft-tissue patches in nasal dorsal augmentation in 66 patients over a 6-year period, which forms the largest European series so far. No complications were observed in either primary (29%) or revision rhinoplasties (71%). PMID- 11437845 TI - A worrying development in the microbiology of otitis externa. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is causing growing concern in hospitals. There has been a steady increase in the number of cases of nosocomial MRSA infections recently and this will no doubt apply to otitis externa, one of the most common ENT infections. The total number of cases of otitis externa presenting to the Accident and Emergency Department over a 3-month period was recorded and the offending microbes cultured and tested for drug sensitivities. Although Pseudomonas aeruginosa was the most frequent organism, 30% of patients grew S. aureus. Of these, 6% (15 patients) were MRSA cultures. The contact histories, antibiotic sensitivities and treatment of these 15 patients were studied. Recommendations as a result of this study include the routine culture and sensitivity in otitis externa and where MRSA is cultured, a full contact history should be elicited and appropriate precautions taken. Specifically, a history of hospital contact should be sought. Treatments used successfully in the treatment of MRSA otitis externa were aural toilet and fucidic acid-betamathasone 0.5% wicks where the organism was gentamycin-resistant (GMRSA), whereas aural toilet with aminoglycoside-steroid drops was sufficient if it was gentamycin sensitive. PMID- 11437846 TI - Specialist Registrars in Otolaryngology: their training programme and career aspirations. AB - The Calman reforms were introduced to Otorhinolaryngology in July 1996 and we are half-way through its first cycle. Further changes may become necessary with expansion of our specialty and because of the need to incorporate super specialization into current training programmes. We felt that it was an appropriate to evaluate these reforms through the eyes of the Specialist Registrars and at the same time to take the opportunity to investigate their career aspirations. In August 1999 a postal questionnaire was sent to all 184 Specialist Registrars in otorhinolaryngology in England and Wales, of which 123 (66.85%) responded. The results are presented in this paper. PMID- 11437847 TI - MIB-1 and p53 expression in radiotherapy-resistant T1aN0M0 glottic squamous cell carcinoma. AB - Radiotherapy of T1aN0M0 glottic carcinoma results in a local control rate of 80 94%. This homogenous group, which is the earliest recognisable invasive malignancy in the head and neck region, provides a 'unique model' for studying possible biological markers of radiosensitivity. p53 and MIB-1 were investigated as possible markers of radiosensitivity in such a group. In all, 107 patients with T1aN0M0 glottic squamous cell carcinoma treated with radiotherapy were identified. Cases not responsive to radiotherapy were compared with matched radiosensitive controls by immunohistochemistry using monoclonal primary antibodies to MIB-1 (n = 18; controls = 10) and p53 (n = 6; controls = 11). No significant difference in p53 expression was noted between the two groups (P = 0.73). A greater MIB-1 expression was found in the radiosensitive group but only a trend towards significance was observed (P = 0.06). MIB-1 is a potential marker of radiosensitivity. A larger multicentre study is required for a more definitive answer. PMID- 11437848 TI - Preliminary results of a pilot study investigating the potential of salivary cortisol measurements to detect occult adrenal suppression secondary to steroid nose drops. AB - Adrenocortical suppression is a well-known risk of systemic steroids, but is thought less likely to occur with topical intranasal corticosteroids. However, the UK Committee on the Safety of Medicines (UKCSM) has expressed concern about the possibility of this complication. We assessed the prevalence of adrenal suppression in patients with rhinitis using intranasal beclomethasone and betamethasone; and the potential value of salivary cortisol as a tool for detecting this complication. Sixty-six patients (38 men: 28 women; mean age 49.6[SD 16.0] years) were prospectively screened for adrenal insufficiency using clinical assessment and salivary cortisol measurements. Abnormalities at this initial screening were confirmed with a Short Synacthen Test (SST). No patient was clinically Cushingoid. All 22 beclomethasone users had normal salivary cortisols. Eleven (25%) of 44 patients using betamethasone had subnormal salivary cortisol levels (mean morning cortisol 2.8[SD 0.9]nmol/l) suggesting adrenal suppression, which was confirmed by an impaired SST in each case. The positive predictive value of salivary cortisol measurements was 100%. Only patients with abnormal salivary cortisols had a SST, so no comment can be made about sensitivity/specificity. Topical betamethasone may produce occult adrenal insufficiency and assessment of adrenal function is recommended in these patients. Measurement of salivary cortisol is a useful, non-invasive and economical test for monitoring patients using intranasal corticosteroids. PMID- 11437849 TI - Hearing evaluation with distortion-product otoacoustic emissions in young patients undergoing haemodialysis. AB - Sensorineural hearing loss is frequently reported in young patients with chronic renal failure having haemodialysis. The effect of a single session of haemodialysis on hearing acuity was assessed prospectively in nine children with end-stage renal disease using pure-tone audiometry (PTA) and distortion-product otoacoustic emissions (DPOAEs). Results were compared with those obtained from nine audiologically normal healthy children also tested with PTA and DPOAEs twice during a 4-h interval. Sensorineural hearing loss of unknown aetiology was found in 55.5% of renal patients, mainly in the higher frequencies. Patients on HD had mean PTA thresholds significantly poorer than those of the control group in the frequency range 1000-12 000Hz (P < 0.05). Their mean DPOAE amplitudes were significantly lower in all frequencies > 1184 Hz (P < 0.05). Furthermore, patients' ears with normal PTA thresholds between 250 and 4000 Hz also had decreased DPOAE amplitudes. No significant changes in PTA thresholds or DPOAE amplitudes were encountered in renal patients before and after a HD session (P > 0.05). Changes in PTA thresholds or DPOAE amplitudes were not significantly different than those in the control group (P > 0.05). In conclusion, sensorineural hearing loss of unknown origin, especially in high frequencies, is frequent in young renal patients under HD and single HD sessions do not seem to alter the hearing acuity of these patients. DPOAEs seem to be more sensitive to incipient cochlear damage than behaviour thresholds in monitoring renal patients. PMID- 11437850 TI - Management of non-tuberculous mycobacterial peri-sialadenitis in children: the Sheffield otolaryngology experience. AB - Non-tuberculous mycobacterial infection (NTM) has been recognized as an important cause of infection in the head and neck in children since 1956. It is important to differentiate NTM from the more serious Mycobacterium tuberculosis (TB) since the management differs significantly. The causative organisms of NTM infection are resistant to the most commonly used anti-tuberculous preparations, though they do show sensitivity to the newer antibiotics such as clarithromycin, ciprofloxacin and azithromycin. Between 1986 and 1997, 15 patients with NTM infection involving the major salivary glands were treated at the Sheffield Children's Hospital Department of Otolaryngology. There were 11 girls and four boys. In all patients the onset of symptoms was between September and April. Resolution occurred in two patients without surgery. The remaining 13 underwent formal surgical exploration with excision of the mass, associated nodes and of the overlying skin if necessary. There were nine parotid explorations. There were no long-term facial nerve deficits as a result of surgery and no recurrence of the disease. Co-operation between the Paediatrician and the Otolaryngologist is important for effective management since NTM may also affect the lungs, soft tissues, bones and joints. Diagnosis relies upon culture, histology, chest radiography, purified protein derivative (PPD) testing, clinical features and skin testing. The use of antibiotics such as azithromycin with ciprofloxacin may be justified while waiting for the results of investigations, especially with small, early lesions, as resolution is possible. In patients who have no history of exposure to TB, are not immunocompromised, have a normal chest radiograph and have a Mantoux test with < 15 mm diameter induration, the treatment should be surgical excision rather than prolonged antibiotic therapy. PMID- 11437851 TI - Publishing trends in otorhinolaryngology from January 1997 to December 1999 in the UK. AB - In the last 10 years there have been many changes in otorhinolaryngology training and academic resources. The Calman reforms were introduced to our speciality in July 1996 and the last decade has also seen the number of professorial chairs in the UK rise from two to 12. One would therefore expect an increase in academic output, in terms of published works, despite the impediments generated by the Calman Training System. A search of eight leading English language otorhinolaryngology journals was carried out from January 1997 to December 1999 looking for articles with British authors. The results were compared with similar research carried out 10 years ago. There has been no major growth in the output of otorhinolaryngological publications from the UK in the last 10 years. PMID- 11437852 TI - Publication rates of scientific papers presented at the Otorhinolarygological Research Society meetings. AB - The aim of this study was to determine the publication rate of scientific papers in peer review journals presented at the Otorhinolarygological Research Society (ORS) meetings from 1978 to 1995 inclusive. The abstracts of the presentations at ORS meetings are published in Clinical Otolaryngology. A MEDLINE search was performed on abstracts presented at ORS meetings from 1978 to 1995 using both authors and key words within the text of the abstract. The publication rate, journal of publication, time to publication, change in contents, change in authors and change in conclusions of abstracts were tabulated. The publication rate for papers presented at ORS meetings from 1978 to 1995 was 69.09%. The average time to publication was 22.5 months. Papers derived from the ORS abstracts were most commonly published in Clinical Otolaryngology (34%) and Journal of Laryngology and Otology (18.64%). The results indicate that nearly 69% of presented material at the biannual ORS meetings eventually get published in peer reviewed journals. This compares favourably with publication rate of other specialities. PMID- 11437853 TI - Prognostic value of electrocochleography in patients with unilateral Meniere's disease undergoing saccus surgery. AB - Although a controversial operation, saccus surgery has continued to be used in patients with severe Meniere's disease in whom hearing preservation is the objective. The aim of this study was to determine if electrocochleography (ECoG) could predict the patients who would be free of vertigo, 2 and 5 years after surgery. Seventy patients with severe Meniere's syndrome underwent ECoG prior to endolymphatic sac surgery. Eighty-four per cent of the patients had an abnormal electrocochleogram. The patients were interviewed 2 and 5 years after surgery. At 2 years, 4/10 (40%) patients with normal ECoG were relieved of dizziness compared to 38/51 (75%) with abnormal ECoG (P < 0.05). At 5 years, 5/8 (63%) patients with normal ECoG were relieved of dizziness compared to 30/30 (100%) with abnormal ECoG (P < 0.001). The patients with a normal ECoG do have less severe measurable hearing impairment (P < 0.05) although they are more likely (P < 0.05) to complain of constant tinnitus. Their history of dizziness is longer and their dizziness more disabling before surgery. In conclusion, patients with a normal ECoG will be less likely to benefit from saccus surgery in the first 5 years. These patients may represent a different nosological entity. PMID- 11437854 TI - Lighting up insulin action. AB - Understanding the mechanism of insulin action remains one of the most important challenges in modern medical biology. Recent advances in cell imaging techniques, increased processing power of computers and the internet, and the introduction of novel fluorescent reagents such as green fluorescent proteins (GFPs) have revolutionized our ability to scrutinize insulin action by time-lapse microscopy at the single-cell level. This article outlines some of the advances made in the authors' laboratory, with particular reference to imaging the movements of the insulin-sensitive glucose transporter, GLUT4, and the generation of phosphoinositide lipids. PMID- 11437855 TI - Prediction of Type 2 diabetes in healthy middle-aged men with special emphasis on glucose homeostasis. Results from 22.5 years' follow-up. AB - AIMS: To study the glucose disappearance rate and fasting blood glucose as predictors of Type 2 diabetes in a 22.5-year prospective follow-up of 1947 healthy non-diabetic men. SUBJECTS AND METHODS: Of a cohort of 2014 Caucasian men, the 1947 who had both fasting blood glucose < 110 mg/dl and an intravenous glucose tolerance test were included. A number of other physiological parameters were also determined at baseline. Multivariate Cox regression analyses were used to investigate the possible significance of the glucose disappearance rate and fasting blood glucose as predictors of Type 2 diabetes. RESULTS: After 22.5 years' follow-up, 143 cases of Type 2 diabetes had developed. Glucose disappearance rate and fasting blood glucose were moderately correlated (r = 0.32). Men in the lowest quartile of glucose disappearance rate and highest quartile of fasting blood glucose had markedly higher diabetes rates than all other men (P < 0.0001). After adjusting for each other, age, diabetes heredity, body mass index, physical fitness, triglycerides, cholesterol and blood pressure (Cox model), both glucose disappearance rate and fasting blood glucose remained major predictors of diabetes CONCLUSIONS: Glucose disappearance rate and fasting blood glucose are, in spite of low intercorrelation, major long-term predictors of Type 2 diabetes in healthy non-diabetic Caucasian men. PMID- 11437856 TI - Abnormal cardiovascular reflex tests are predictors of mortality in Type 2 diabetes mellitus. AB - AIMS: To determine whether diabetic autonomic neuropathy is an important factor contributing to mortality in Type 2 diabetes mellitus. METHODS: Between 1989 and 1993, 431 men and 181 women with Type 2 diabetes were given diabetic autonomic neuropathy cardiovascular reflex (CVR) tests. These subjects were followed for the subsequent 5--9 years to assess mortality rates. RESULTS: The prevalence rate of abnormal CVR tests was 46.1% in patients with the history of diabetes less than 5 years and up to 69.4% when the history of diabetes exceeded 20 years. During the follow-up period from 1 January 1989 to 31 December 1997 (mean 7.7 years), a total of 135 participants died. The 8-year survival rate for patients with abnormal CVR tests was 63.6% in males and 76.4% in females, compared with 80.9 and 93.3% for patients with normal CVR tests. The results were grouped as: group 1, normal CVR tests without postural hypotension (PHT); group 2, normal CVR tests with PHT; group 3, abnormal CVR tests without PHT; and group 4, abnormal CVR tests with PHT. The 8-year survival rate was 85.4% in group 1, 80.9% in group 2, 74.5% in group 3 and 61.1% in group 4. CONCLUSION: Type 2 diabetic patients with abnormal CVR tests may have increased mortality, and those combined with postural hypotension have higher mortality than those without. Abnormal CVR tests may be important predictors of mortality in Type 2 diabetes mellitus. PMID- 11437857 TI - Glycated haemoglobin assays. Approaches to standardization of results. AB - AIMS: To compare different approaches to DCCT standardization of glycated haemoglobin (HbA(1c)) results. METHODS: In the first part of the study seven laboratories in the Mersey area participated, using a variety of methods of measurement. The approaches used were the Standardization Initiative for Glycated Haemoglobin Scheme (SIGH) using fresh blood samples and the Wales External Quality Assurance Scheme (WEQAS) using both fresh and lyophilized blood samples. Additional studies performed in the Royal Liverpool University Hospital (RLUH) laboratory involved: (i) re-calibration of the HPLC analyser using lyophilized blood samples supplied by the manufacturer and (ii) the use of fresh blood samples from the UK National External Quality Assurance Scheme (UKNEQAS). RESULTS: Inter-laboratory analytical coefficients of variation following DCCT alignment showed little change. The effect on percentage bias was more marked and was independent of the level of HbA(1c). Comparing DCCT-aligned HbA(1c) results from 3902 diabetic patients attending the RLUH, indicated that there was a statistically significant difference (P < 0.0001) between the corrected results produced by the five schemes. The effect of DCCT alignment on reported HbA(1c) values using the different approaches was variable resulting in an apparent increase in the number of diabetic patients with poor glycaemic control (HbA(1c) values > 7.0%) in all cases. CONCLUSION: The method of HbA1c standardization used determines the degree of correction required and a consensus approach is recommended. Implementation will assist extrapolation from research-based evidence to local practice. Associated changes to patient management must come under the jurisdiction of the physicians responsible for the diabetic service. PMID- 11437858 TI - Intra-urban differences in the prevalence of the metabolic syndrome in southern India -- the Chennai Urban Population Study (CUPS No. 4). AB - AIMS: To assess the influence of socioeconomic status on the prevalence of the metabolic syndrome in an urban south Indian population in Chennai. METHODS: The Chennai Urban Population Study is an epidemiological study involving two residential colonies in Chennai (formerly Madras) in south India representing the middle and lower income groups. All individuals > or = 20 years of age living in the colonies were invited to participate in the study. Of the total 1399 eligible subjects, 1262 individuals (479 belonging the middle income group colony and 783 from the low income group colony) participated in the study. The overall response rate was 90.2%. The main outcome measures were the prevalence rates of the various components of the metabolic syndrome. RESULTS: There were significant differences in the socioeconomic status and lifestyle of the inhabitants of the two areas. The mean monthly income of the Tirumangalam (middle income) group (Rs8075 +/- 3859) was significantly higher than the T. Nagar (low income) group (Rs1399 +/- 916). The dietary profile of the middle income group showed higher intake of calories, fat and sugar compared to low income group (P < 0.001). The age-standardized prevalence rates of the various components of the metabolic syndrome were significantly higher in the middle compared to the low income group - diabetes (12.4 vs. 6.5%), impaired glucose tolerance (7.5 vs. 2.9%), hypertension (14.9 vs. 8.4%), obesity (males 38 vs. 13.4%, females 33.1 vs. 24.2%), hypercholesterolaemia (24.2 vs. 14.2%) and hyperinsulinaemia (16.7 vs. 6.6%) P < 0.001). Although the prevalence of coronary artery disease and hypertriglyceridaemia were higher in the middle income group, the differences did not reach statistical significance. The relative odds ratio for diabetes and impaired glucose tolerance increased significantly with increase in income while hypercholesterolaemia, hypertriglyceridaemia, hypertension and coronary artery disease showed no significant changes. Logistic regression analysis revealed that geographical area (higher social class) had a strong association with the components of the metabolic syndrome even after inclusion of other risk factors like age and body mass index in the model. CONCLUSIONS: Significant differences exist in the prevalence of various components of the metabolic syndrome even within an urban environment and this appears to be influenced by socioeconomic status. PMID- 11437859 TI - Circulating plasma vascular endothelial growth factor and microvascular complications of type 1 diabetes mellitus: the influence of ACE inhibition. AB - AIMS: To determine whether circulating plasma vascular endothelial growth factor (VEGF) is elevated in the presence of diabetic microvascular complications, and whether the impact of angiotensin-converting enzyme (ACE) inhibitors on these complications can be accounted for by changes in circulating VEGF. METHODS: Samples (299/354 of those with retinal photographs) from the EUCLID placebo controlled clinical trial of the ACE inhibitor lisinopril in mainly normoalbuminuric non-hypertensive Type 1 diabetic patients were used. Albumin excretion rate (AER) was measured 6 monthly. Geometric mean VEGF levels by baseline retinopathy status, change in retinopathy over 2 years, and by treatment with lisinopril were calculated. RESULTS: No significant correlation was observed between VEGF at baseline and age, diabetes duration, glycaemic control, blood pressure, smoking, fibrinogen and von Willebrand factor. Mean VEGF concentration at baseline was 11.5 (95% confidence interval 6.0--27.9) pg/ml in those without retinopathy, 12.9 (6.0--38.9) pg/ml in those with non-proliferative retinopathy, and 16.1 (8.1--33.5) pg/ml in those with proliferative retinopathy (P = 0.06 for trend). Baseline VEGF was 15.2 pg/ml in those who progressed by at least one level of retinopathy by 2 years compared to 11.8 pg/ml in those who did not (P = 0.3). VEGF levels were not altered by lisinopril treatment. Results were similar for AER. CONCLUSIONS: Circulating plasma VEGF concentration is not strongly correlated with risk factor status or microvascular disease in Type 1 diabetes, nor is it affected by ACE inhibition. Changes in circulating VEGF cannot account for the beneficial effect of ACE inhibition on retinopathy. PMID- 11437860 TI - Factors affecting the use of dietetic services by patients with diabetes mellitus. AB - AIMS: Dietary counselling is vital for patients with diabetes, yet many do not access professional advice. This study investigated the use of dietetic services by patients with diabetes within Sunderland Health District. METHODS: Diabetic patients were identified from the laboratory module of the Hospital Information System (HIS). Eight thousand five hundred and ninety-seven patients were identified, and cross referenced to dietetic records, producing a comprehensive database for analysis. RESULTS: Only 58.8% of the sample had dietetic records. Those with records were more likely to be: older (63.8 vs. 61.7 years, P < 0.01); male (52.6% vs. 46.8%, P < 0.05); accessing hospital diabetes services (P < 0.001); to have lipids and renal function checked (both P < 0.001); and to have poor blood glucose control (HbA(1c) > 7.5% to 58.7% vs. 29.9%, P < 0.001). Frequent attenders were most likely to be women, those with poor control, and those with shorter duration of diabetes (P < 0.001). In a logistic regression model, duration of diabetes, insulin treatment, obesity and hyperlipidaemia were the main determinants of increased use. CONCLUSIONS: This study confirmed that many patients with diabetes do not make use of professional dietetic services, and has highlighted some of the routinely documented characteristics that are associated with use of dietetic services. There appears to be scope to improve uptake of dietetic services by patients with diabetes, and to investigate further individual factors that affect access and attendance. PMID- 11437861 TI - Evaluation of a hospital diabetes specialist nursing service: a randomized controlled trial. AB - AIMS: To evaluate the effectiveness and cost implications of a hospital diabetes specialist nursing service. METHODS: We conducted a prospective, open, randomized, controlled trial of standard in-patient care for adults with diabetes, with and without the intervention of a diabetes specialist nursing (DSN) service. The setting was a single UK university hospital. SUBJECTS: were unselected patients referred to the hospital DSN service. Primary outcome measures were length of hospital stay and patterns of readmission (frequency and time to first readmission). Secondary outcome measures were subjects' diabetes related quality of life, diabetes knowledge score, satisfaction with treatment, and GP and community care contacts following discharge. Costs were estimated from the hospital and published sources. RESULTS: Median length of stay was lower in the intervention group (11.0 vs. 8.0 days, P < 0.01). Readmission rates were the same in the two groups (25%), and mean time to readmission was similar in the two groups, although slightly less in the control group (278 vs. 283 days, P = 0.80). The cost per patient for nursing input was 38.94 pounds sterling. However, when the reduced length of stay was accounted for, the intervention produced a mean cost per admission of 436 ponds sterling lower than that of the control group (P = 0.19). Patients in the intervention group were more knowledgeable regarding their diabetes and more satisfied with their care. CONCLUSIONS: Diabetes specialist nurses are potentially cost saving by reducing hospital length of stay (LOS). There was no evidence of an adverse effect of reduced LOS on re admissions, use of community resources, or patient perception of quality of care. PMID- 11437862 TI - Effect of troglitazone on urinary albumin excretion and serum type IV collagen concentrations in Type 2 diabetic patients with microalbuminuria or macroalbuminuria. AB - AIMS: Troglitazone, a newly developed thiazolidinedione derivative, has been shown to ameliorate microalbuminuria in diabetic animal model and in human diabetic nephropathy in short-term studies. The aim of the present study was to determine whether troglitazone or sulphonylurea affect micro- albuminuria, macroalbuminuria, or serum type IV collagen concentrations in patients with diabetic nephropathy. METHODS: We studied 32 normotensive patients with type 2 diabetes mellitus associated with microalbuminuria (n = 16) or macroalbuminuria (n = 16) and 20 healthy controls. The patients were randomly assigned to one of two groups: those treated with glibenclamide (5.0 mg/day) (n = 8) and those treated with troglitazone (400 mg/day) (n = 8). They received the drug regimen for 12 months. Serum type IV collagen was measured with sandwich enzyme immunoassay. RESULTS: Type IV collagen concentrations in macroalbuminuric patients were higher than those in microalbuminuric patients (P < 0.05) and healthy controls (P < 0.01). Troglitazone reduced urinary albumin excretion (UAE) in micro-albuminuric patients from 126 microg/min (range 58--180 microg/min) to 42 microg/min (range 14--80 microg/min) (P < 0.01) and also reduced serum type IV collagen levels gradually at 3, 6 and 12 months after treatment (P < 0.05). However, glibenclamide did not affect UAE and type IV collagen levels in micro- albuminuric diabetes patients. In addition, neither troglitazone nor gliben- clamide changed UAE and type IV collagen levels in macroalbuminuric patients. CONCLUSIONS: These data suggest that troglitazone is an effective treatment for renal injury in patients with early diabetic nephropathy. PMID- 11437863 TI - Effects of preventative footwear on foot pressure as determined by pedobarography in diabetic patients: a prospective study. AB - AIMS: This study analysed the effects of specially manufactured insoles on foot pressures in diabetic patients during a 1-year prospective observation period. METHODS: We studied 81 type 2 diabetic patients without foot lesions. Using pedobarography three different regions of interest were examined: maximum peak pressure (MPP) of the total foot area, heel region and head of metatarsal bone I III. Eighteen patients with high risk pressure (MPP of total foot 474 +/- 183 kPa; heel region 278 +/- 147 kPa, metatarsal 389 +/- 222 kPa) received optimal insole support. Sixty-three patients as a control group (MMP of total foot 367.7 +/- 157 kPa; heel 263.1 +/- 127 kPa, metatarsal 339.9 +/- 171 kPa) received conventional footwear. RESULTS: After insole support a 30% pressure reduction of total foot MMP (474 +/- 183 kPa vs. 290 +/- 106 kPa) was achieved in the treatment group. After 6 months (324 +/- 127 kPa) and 1 year (380 +/- 190 kPa) a pressure reduction was found. Between the 6- and 12-month controls plantar pressures again increased. In the control group a significant increase of all peak pressures occurred. CONCLUSIONS: Early insole support is successful in reducing plantar pressure. A repeated adjustment should be performed every 6 months to prevent foot pressure increases. The comparison of foot pressure development between the two groups showed constant levels in the treatment group. In the control group a marked increase of the pressure values was found. Identification and subsequent support of patients with high ulceration risk may help to reduce the high amputation rate. PMID- 11437864 TI - Sodium carboxyl-methyl-cellulose dressings in the management of deep ulcerations of diabetic foot. AB - AIMS: To test the safety and effectiveness of carboxyl-methyl-cellulose dressing (Aquacel; ConvaTec, UK) in the management of deep diabetic foot ulcers, a group of consecutive out-patients attending the foot clinic of the Department of Metabolic Diseases was studied. METHODS: Patients were selected according to the following inclusion criteria: a foot ulcer deeper than 1 cm for > 3 weeks, good peripheral blood supply (palpable peripheral pulses or ABPI > 0.9). Exclusion criteria were as follows: active infection, as evident from clinical signs (purulent discharge, redness, swelling, tenderness) and confirmed by culture exams, plasma creatinine > 2 mg/dl, recent episodes of ketoacidosis, malignancies, and any therapy or pathology which might interfere with the healing process. Twenty patients were enrolled in the study and having obtained their informed consent, their lesions were surgically debrided with the complete elimination of all necrotic tissue and debris up to the bleeding healthy tissue; then ulcers were staged and measured, and patients were randomly assigned to two different treatment groups. Patients in group A were dressed with saline moistened gauze, while patients in group B were dressed with Aquacel according to the manufacturer's instructions. All patients in both groups received special post-operative shoes (Podiabetes; Zeno Buratto, Treviso, Italy) and crutches until complete re-epithelialization. Ulcers were all left to heal by secondary intent. After 8 weeks patients were blindly evaluated for: the rate of reduction of lesional volume (RLV), rate of granulation tissue (GT), number of infective complications (IC). Intralesional (ILTC) and perilesional (PLTC) temperatures were also recorded with a thermocouple surface digital thermometer, and the difference between the two values (Delta TC) was calculated. Healing time (HT, days), was then compared between the two groups. Data were compared by analysis of variance (ANOVA), linear regression, Kaplan-Meier survival analysis and Fisher's exact test. RESULTS: HT was significantly shorter in Group B than in Group A (P < 0.001). RLV was significantly (P < 0.01) higher in Group B patients compared with Group A, as well as GT (P < 0.05). IC were in 1/10 Group B and in 3/10 Group A (P = 0.582). In addition, both ILTC and Delta TC were higher in Group B compared with Group A ones (P < 0.01). CONCLUSIONS: Carboxyl-methyl cellulose dressings were shown to be safe, effective and well tolerated in the management of non-ischaemic, non-infected deep diabetic foot ulcers. PMID- 11437865 TI - Squamous cell carcinoma developing in necrobiosis lipoidica. AB - We describe a case of squamous cell carcinoma arising in long-standing necrobiosis lipoidica in a type 1 female diabetic patient. The tumour and the skin lesion were successfully excised and repaired with full thickness skin graft. The development of squamous cell carcinoma in association with this skin disorder is rare (only four cases reported in literature since 1966), but should be considered in chronic, non-healing and recalcitrant ulcers developing within areas of necrobiosis lipoidica. PMID- 11437866 TI - Fulminant autoimmune Type 1 diabetes during interferon-alpha therapy: a case of Th1-mediated disease? AB - AIM: A case of autoimmune Type 1 diabetes with some unique characteristics developing in a 29-year-old male during treatment with interferon-alpha (IFN alpha) for chronic hepatitis C virus (HCV) hepatitis is reported. PATIENT AND METHODS: In this patient IFN-alpha treatment was well tolerated and successful in the cure of hepatitis with eradication of HCV infection within 3 months, but at 8.5 months Type 1 diabetes appeared and insulin therapy was started and maintained thereafter. HLA class II molecular typing was determined and retrospective measurement of islet cell (ICA), glutamate decarboxylase (GADA), tyrosin phosphatase IA-2 (IA-2A) and insulin (IAA) antibodies was performed in serum samples obtained before and at 0.5, 1, 2, 3, 4, 5, 6, 8.5, 10 and 13 months after the beginning of IFN-alpha treatment. RESULTS: Complete HLA class II typing was consistent with homozygosity for the HLA DRB *03011, DQA1 *0501, DQB1 *0201 haplotype. All autoantibodies were undetectable prior to IFN-alpha therapy and remained undetectable up to 6 months of treatment; at 8.5 months, at the time of diabetes onset, ICA were detectable at low titre while GADA were present at high titre. Both ICA and GADA persisted at high levels in subsequent samples. IA-2A remained undetectable in all serum samples, while IAA appeared only after treatment with exogenous insulin. DISCUSSION: This appears to be a case of autoimmune Type 1 diabetes induced by IFN-alpha treatment and developing on a predisposed genetic background with an unusually rapid development of the autoimmune process as reflected by the absence of detectable autoantibodies up to 2.5 months prior to disease onset. In this example of fulminant Type 1 diabetes a pathogenic process unbalanced towards a Th1-mediated autoimmune response is hypothesized. Diabet. Med. 18, 329-332 (2001) PMID- 11437867 TI - Approaching St Vincent. Working toward the St Vincent targets. PMID- 11437868 TI - Association of the T14709C mutation of mitochondrial DNA with maternally inherited diabetes mellitus and/or deafness in an Italian family. PMID- 11437869 TI - Long hypoglycaemic periods detected by subcutaneous continuous glucose monitoring in toddlers and pre-school children with diabetes mellitus. PMID- 11437870 TI - Delta sleep-inducing peptide. PMID- 11437871 TI - Monitoring in neuroanaesthesia: update of clinical usefulness. AB - The aim of specific monitoring in neuroanaesthesia is to detect, as quickly as possible, intraoperative ischaemic insults so that the brain and the spinal cord may be protected from harmful and frequently inevitable events due to the type of surgery, patient positioning, haemodynamic changes or any intercurrent event. New monitors are being introduced into the operating theatre, but only a few are considered to be an absolute standard of care in neurosurgery, e.g. facial nerve monitoring for surgery of acoustic neuromas and recording of evoked potentials during repair of scoliosis. In the past decade, new monitoring devices have moved from the experimental stage to the operating theatre and although most are still in a phase of technological development and/or definition of their field of applicability they are being used as guides for clinical practice in those instances where cerebral well-being might be impaired. The metabolic consequences of hyperventilation, pharmacological electroencephalogram burst suppression, hypothermia, etc. can now be assessed in the operating theatre. Non-invasive monitoring is being rapidly integrated into our daily work because of its lack of secondary effects. Nevertheless, each new development is regarded as an addition rather than as a substitute for existing equipment. The perfect combination of monitors to provide essential information during an individual surgical procedure to influence a better patient outcome, is still uncertain and needs extensive clinical research. PMID- 11437872 TI - Pharmacodynamic interaction of nitrous oxide with sevoflurane, desflurane, isoflurane and enflurane in surgical patients: measurements by effects on EEG median power frequency. AB - BACKGROUND AND OBJECTIVE: This study investigates the interaction of sevoflurane and nitrous oxide on EEG median power frequency of 2.5 Hz during surgery. METHODS: Sevoflurane concentrations required for electroencephalographic median power frequency between 2 and 3 Hz were measured in 25 patients during gynaecological laparotomies. Nitrous oxide was randomly administered at 0, 20, 40, 60 and 75 vol%, subsequently two different concentrations in each patient. The data were analysed using isobolographic analysis together with previously published data on nitrous oxide-isoflurane, -enflurane, or -desflurane interaction. RESULTS: The interaction is described by the equation: C volatile anaesthetic/C0 volatile anaesthetic + C N2O/C0 N2O=1 (C is the concentrations for a drug combination to achieve the desired effect; C0 is the concentration for single drug use). The parameters are C0 isoflurane=1.11 vol% (95% CI 1.03-1.19), C0 enflurane=1.64 (1.52-1.77), C0 desflurane=5.31 (4.92-5.73), C0 sevoflurane=2.12 (1.96-2.29), C0 N2O=174 (153-202). These parameters decrease by 6% (2.5-10) for every 10 years of patients' age > 40 years. CONCLUSIONS: The interaction is compatible with additivity. The potency of nitrous oxide to substitute the volatile anaesthetics is less than anticipated from previously reported MAC values. PMID- 11437873 TI - A comparison of five solutions of local anaesthetics and/or sufentanil for continuous, postoperative epidural analgesia after major urological surgery. AB - BACKGROUND AND OBJECTIVE: The aim of the present study was to compare and assess the quality of analgesia, the safety and the side-effects after the use of a continuous, thoracic epidural infusion of sufentanil (5 microg h(-1)), 0.25% bupivacaine (10 mL h(-1)), 0.2% ropivacaine (10 mL h(-1)) alone or in combination in patients who had undergone major urological surgery. This prospective, randomized, double-blinded study investigated the efficacy of thoracic epidural infusions after major urological surgery. METHODS: Patients received a 72-h continuous infusion (10 mL h(-1)) of 0.25% bupivacaine (B), 0.2% ropivacaine (R), 0.25% bupivacaine with 0.5 microg mL(-1) sufentanil (BS), 0.2% ropivacaine with 0.5 microg mL(-1) sufentanil (RS) or 0.5 microg mL(-1) sufentanil only (S). The analysis included 109 patients. RESULTS: The mean visual analogue scale (VAS) scores for pain were highest in the groups R and S (P < 0.001). The PaCO2 values were significantly higher in the groups RS and S (P = 0.003). Motor block occurred more frequently in the groups B and BS than in the other groups (P < 0.001). Sedation, nausea and pruritus were more common in the groups that received sufentanil. CONCLUSIONS: A continuous, epidural infusion with these drugs was safe and effective in our patients. The combination of 0.2% ropivacaine plus sufentanil appeared preferable because of the low incidence of motor block. PMID- 11437874 TI - Minimal-flow anaesthesia with controlled ventilation: comparison between laryngeal mask airway and endotracheal tube. AB - BACKGROUND AND OBJECTIVE: Minimal- and low-flow anaesthesia (fresh gas flow below 1 L min(-1)) provide many advantages, including reduced cost, conservation of body heat and airway humidity. An airtight seal is essential between the airway device and the airway of the patient. Therefore, we investigated whether the airtight seal created by a laryngeal mask airway allows controlled ventilation of the lungs when the fresh gas flow is reduced to 0.5 L min(-1) and compared this with an endotracheal tube. METHODS: In a prospective clinical study, 207 patients were managed using a laryngeal mask or an endotracheal tube. After intravenous induction of anaesthesia and 15 min of high fresh gas flow, the flow was reduced to 0.5 L min(-1). The breathing system was monitored for airway leaks, and the patients were assessed for complications after airway removal and postoperative discomfort. RESULTS: Both the laryngeal mask and endotracheal tube allowed fresh gas flow reduction to 0.5 L min(-1) in 84.7% and 98.3% of cases respectively (small leaks: 12% laryngeal mask, 1.7% endotracheal tube). Three patients with the laryngeal mask (3.3%) had airway leaks that were too large to permit any reduction in the fresh gas flow. CONCLUSIONS: The use of the laryngeal mask airway was more likely to be associated with a gas leak than use of an endotracheal tube; however, if modern anaesthesia machines and monitors are used, in 96.7% of the patients managed with a laryngeal mask a reduction in the fresh gas flow to 0.5 L min(-1) was possible. The incidence of coughing and postoperative complaints (sore throat, swallowing problems) was higher after use of an endotracheal tube. PMID- 11437875 TI - Effects of temperature on partial thromboplastin time in heparinized plasma in vitro. AB - BACKGROUND AND OBJECTIVE: Perioperative hypothermia has been found to impair the coagulation cascade and to increase blood loss and transfusion requirements. The effect of concomitant in vitro heparinization on coagulation during hypo- and hyperthermic conditions has not been well defined. METHODS: In the present study, activated partial thromboplastin time was examined in vitro at 33 degrees C, 35 degrees C, 37 degrees C, 39 degrees C and 41 degrees C in normal human plasma in response to unfractionated heparin. RESULTS: Hypothermia 5 points) lower mean scores on seven of the eight SF-36 domains and both the mental and physical summary scores of the SF-36. CONCLUSIONS: The results of this study suggest that patients with transformed migraine have a lower health-related quality of life than patients with migraine. These findings indicate that the headache chronicity associated with transformed migraine has a significant influence on quality of life. The results highlight the importance of effective management of headaches to avoid the progression of migraine to the more disabling transformed migraine. PMID- 11437894 TI - Event-related potentials in posttraumatic headache. AB - OBJECTIVES: To assess impairment of cognitive functions occurring in patients with posttraumatic headache as a consequence of a minor cranial trauma in the absence of organic damage involving the central nervous system. BACKGROUND: The term posttraumatic syndrome defines a stereotypic set of symptoms following traumatic brain injury that are subjective and varied. A deficit of cognitive function and impairment of the rapid processes of learning, attention, and short term memory have frequently been identified. Moreover, headache is the most frequent symptom reported by the patients. Due to the nature of the symptoms, a great limitation in defining the posttraumatic syndrome is represented by the lack of methods and diagnostic tools that allow quantification of the subjective disturbances and evidence of the signs indicative of central nervous system involvement in this pathological condition. METHODS: Twenty-five subjects (16 women, 9 men; mean age, 28 +/- 9 years) were examined between 3 and 6 months after the traumatic event. The P300 event-related potential was recorded by an odd-ball paradigm with an acoustic modality. The patients underwent electroencephalography and brain stem auditory evoked potentials; magnetic resonance imaging was performed to exclude the presence of cerebral lesions. RESULTS: The mean latency of P300 was increased in both central electrodes (Cz and Pz) in patients with posttraumatic syndrome compared with controls (P<.001); assuming the value of mean +/- 2 SD was the cutoff point between normal and abnormal results, the P300 latency results were altered in 13 patients (52%). In the patient group, a significant correlation was demonstrated between Zung Depression Scale score and P3 and N2 wave latencies (r = 0.54, P <.004; r = 0.56, P<.003) and between Zung Anxiety Scale scores and P3 wave latencies (r = 0.46, P<.02). CONCLUSIONS: These data suggest the usefulness of the P300 event-related potential in evaluating cognitive disturbances in patients affected by posttraumatic syndrome. Alteration of cognitive potential in such patients, even in the absence of lesions detectable by neuroimaging, indicate the functional impairment of specific cerebral areas that can occur after a traumatic event. PMID- 11437895 TI - Sleep disturbances after whiplash injury: objective and subjective findings. AB - OBJECTIVE: A controlled objective and subjective evaluation of sleep in a consecutive series of persons who had had a whiplash injury, shortly following the injury and 3 to 5 months later. METHODS: In whiplash-injured subjects and healthy controls, sleep characteristics were monitored objectively throughout the night by means of an actigraph. The following morning, participants in both groups filled out a prestructured "sleep log" that conveyed their subjective impressions as to how they had slept. The correlation between objectively measured sleep characteristics and the symptoms and physical findings, respectively, resulting from whiplash injury were also examined. RESULTS: Actigraphic monitoring did not reveal a significant group difference between whiplash-injured subjects and controls with respect to any of the sleep characteristics recorded. In self-maintained "sleep logs," however, the whiplash injured subjects reported a significantly prolonged sleep latency and significantly impaired sleep quality compared with controls (P<.001 and P<.04, respectively). In whiplash-injured subjects, the number of arousals was positively correlated with the number of symptoms and with the number of physical signs of whiplash injury (P<.01 and P<.001, respectively). Sleep efficiency was inversely correlated with the number of injury-related findings on physical examination (P<.009). CONCLUSION: Sleep, as monitored by actigraph in whiplash injured subjects, was not adversely affected by whiplash injury. The subjective impressions of the subjects, as recorded in "sleep logs," suggest the opposite conclusion. A significant correlation exists between certain symptoms and signs of whiplash injury and abnormalities in given sleep characteristics as detected by actigraph. PMID- 11437896 TI - Sumatriptan can alleviate headaches due to carotid artery dissection. AB - Cluster headaches can be mimicked by a spontaneous carotid artery dissection. We report a 45-year-old man with a spontaneous carotid artery dissection whose unilateral headache responded to sumatriptan. An oral dose of 50 mg of sumatriptan relieved 90% of the pain after 2 hours. A second dose the next day achieved similar results within 4 hours. The diagnosis of dissection was made later by magnetic resonance angiogram and conventional angiography. This case illustrates that a positive response to a triptan can not be used to distinguish the first attack of cluster headache from a carotid artery dissection. PMID- 11437897 TI - Remitting form of hemicrania continua with seasonal pattern. AB - Hemicrania continua is a primary headache syndrome characterized by a continuous, unilateral headache that is completely responsive to indomethacin. Hemicrania continua exists in continuous and remitting forms. Ten cases of the remitting form have been reported, none of which have had a seasonal pattern. We report a patient with remitting hemicrania continua with a clear seasonal predilection. PMID- 11437898 TI - Idiopathic intracranial hypertension and hemophilia A. AB - OBJECTIVE: A patient with hemophilia A and long-standing recurrent symptoms of idiopathic intracranial hypertension is described. During his relapses, he experienced headache, and attention and language disturbance, but no visual symptoms. BACKGROUND: Hemophilia A is a rare inherited coagulation disorder secondary to factor VIII deficiency. Idiopathic intracranial hypertension has been reported in association with prothrombotic conditions and iron deficiency anemia, but not in patients with hemophilia A. Recurrent or chronic headache is not a typical symptom of hemophilia, but headache is a presenting sign of intracranial bleed in persons with hemophilia. METHODS: Medical history review, clinical neurologic examination, brain magnetic resonance imaging, computed head tomography, and electroencephalogram were performed. RESULTS: Neurologic examination revealed bilateral papilledema during relapses of idiopathic intracranial hypertension. Multiple lumbar punctures preceded by the intravenous administration of factor VIII early in the course of the illness confirmed the presence of elevated cerebrospinal fluid pressures and absence of subarachnoid blood. He had no complications from lumbar punctures. Initial electroencephalograms showed background slowing but later normalized. Magnetic resonance imaging of the brain and computerized tomography of the head were normal. Relapses of idiopathic intracranial hypertension were eventually controlled with the administration of acetazolamide. CONCLUSION: Idiopathic intracranial hypertension may develop in patients with hemophilia A in the absence of visual symptoms. Therapeutic and diagnostic lumbar punctures were safe to perform on this patient, following the administration of factor VIII. PMID- 11437899 TI - Stabbing headache as the presenting manifestation of intracranial meningioma: a report of two patients. AB - Stabbing headache may occur as a primary headache disorder, may accompany other primary headaches, or, in rare instances, may be associated with secondary headache syndromes. We report two patients with intracranial meningioma in whom stabbing headache was the heralding symptom. Headache remitted promptly following surgical resection. These cases stress the importance of excluding underlying pathology in patients presenting with new-onset stabbing headache. PMID- 11437900 TI - Exploding head syndrome. PMID- 11437901 TI - Episodic vertigo and migraine. PMID- 11437902 TI - Re: "Botulinum toxin type A as a migraine preventive treatment" (Silberstein S, Mathew N, Saper J, Jenkins S, for the BOTOX Migraine Clinical Research Group. Headache. 2000;40:445-450). PMID- 11437903 TI - Re: "Botulinum toxin type A as a migraine preventive treatment" (Silberstein S, Mathew N, Saper J, Jenkins S, for the BOTOX Migraine Clinical Research Group. Headache. 2000;40:445-450). PMID- 11437904 TI - Effect of rizatriptan in the spectrum of headache. PMID- 11437905 TI - Paroxysmal hemicrania-tic syndrome. PMID- 11437906 TI - The elusive role of thrombin in migraine. PMID- 11437907 TI - We need better preventative medications. PMID- 11437908 TI - Induction of a migraine aura in a patient suffering from migraine without aura. PMID- 11437909 TI - Headache in the colonies: the Mayflower virus? PMID- 11437910 TI - Cytochrome P450s CYP6D3 and CYP6D1 are part of a P450 gene cluster on autosome 1 in the house fly. AB - The P450 monooxygenases of insects are important in the metabolism of numerous endogenous and exogenous compounds. However, identity of the P450 isoform(s) involved in these reactions is rarely known. A critical first step in the identification of important P450s is the cloning and sequencing of their genes. Toward this goal we report the genomic sequence of a new cytochrome P450, termed CYP6D3, from the house fly, Musca domestica. CYP6D3 is part of a P450 gene cluster located on chromosome 1 and is located upstream of a related gene, CYP6D1. The similar genetic structures of CYP6D3 and CYP6D1 (5 exons and 4 introns of similar length) suggest one of these genes may have been the result of a duplication event. The CYP6D3 deduced amino acid sequence indicates a protein with 518 amino acids and a molecular weight of 59.3 kDa. The CYP6D3 protein is most similar to house fly CYP6D1 (78%) and Cyp6D2 (56%) from Drosophila melanogaster. The deduced amino acid sequences of CYP6D3 and CYP6D1 are identical at the Helix I and heme binding regions. PMID- 11437911 TI - Comparative linkage map development and identification of an autosomal locus for insensitive acetylcholinesterase-mediated insecticide resistance in Culex tritaeniorhynchus. AB - A comparative linkage map for Culex tritaeniorhynchus was constructed based on restriction fragment length polymorphism markers using cDNAs from Aedes aegypti. Linear orders of marker loci in Cx. tritaeniorhynchus were identical to Culex pipiens wherein chromosomes 2 and 3 reflect whole-arm rearrangements compared to A. aegypti. However, the sex determination locus in Cx. tritaeniorhynchus maps to chromosome 3, in contrast to Cx. pipiens and Ae. aegypti where it is located on chromosome 1. Our results indicate that insensitive acetylcholinesterase (AChE) mediated organophosphate resistance is controlled by a single major gene (AChE) on chromosome 2, while the AChE structural gene (Ace) is located on chromosome 1. No evidence for a second Ace gene was observed, even under very low stringency hybridization conditions. PMID- 11437912 TI - Sequence and organization of the mitochondrial genome of the Chagas disease vector, Triatoma dimidiata. AB - The 17 019 bp mitochondrial genome of Triatoma dimidiata is composed of thirteen protein coding sequences, twenty-two tRNAs, small and large ribosomal units, and a control region. The gene order and orientation are identical to that of Drosophila yakuba. The nucleotide composition is biased toward adenine and thymine (69.5% A + T). The 2.1 kb putative control region, known as the A + T rich region in most insects, has an A + T bias of 66%, but contains a 400 bp sequence that is 77.5% A + T and two other distinct regions: (1) one with a lower A + T bias (60.1%) and (2) a region of eight tandem repeat units. The identified 1.4 kb nuclear copy of mitochondrial sequences encompasses the string of Gs and the beginning of the cytochrome c oxidase 1 gene but lacks the 1.8 kb region spanning the eight tandem repeats and the 5' end of the NADH dehydrogenase subunit II gene. PMID- 11437913 TI - Beta-integrin of Anopheles gambiae: mRNA cloning and analysis of structure and expression. AB - We have isolated an mRNA encoding a beta integrin subunit of the malaria mosquito Anopheles gambiae. Our analysis predicts a protein that is very similar to betaPS, the fruitfly orthologue. The gene is expressed during all developmental stages and it is found in all body parts, including the midgut. Finally, the expression of the gene does not seem to be modulated during blood meals, except for a substantial increase 48 h posthaematophagy, when digestion is nearly complete. PMID- 11437914 TI - Microsatellite loci are not abundant in all arthropod genomes: analyses in the hard tick, Ixodes scapularis and the yellow fever mosquito, Aedes aegypti. AB - Plasmid libraries enriched for microsatellites were generated in the tick, Ixodes scapularis and in the mosquito Aedes aegypti. Libraries were enriched for genomic DNA containing (AC)n, (AG)n, (ATG)n, (CAG)n, (TAG)n, (AAT)n, (CTGY)n or (GATA)n motifs. Clones containing each motif were sequenced in both species for PCR primer design. In I. scapularis, most primers amplified a single locus and alleles varied in the number of microsatellite repeats and segregated as codominant markers. In contrast (AC)n, (TAG)n and (GATA)n microsatellite loci extracted from Ae. aegypti appeared to be members of multigene families. A primer pair designed to amplify a particular TAG locus instead amplified many independently segregating loci, some of which did not contain TAG microsatellites. Alleles at the TAG loci segregated as dominant markers and there was limited evidence for length variation among alleles. These results suggest that microsatellite loci are not universally abundant in arthropod genomes nor do alleles always segregate as codominant markers. PMID- 11437915 TI - Evolutionary change in the structure of the regulatory region that drives tissue and temporally regulated expression of alcohol dehydrogenase gene in Drosophila funebris. AB - The Adh locus of Drosophilidae is organized as a single gene transcribed from two spatially and temporally regulated promoters except in species of the repleta group, which have two single promoter genes. Here we show that in Drosophila funebris the Adh gene is transcribed from a single promoter, in both larva and adult, with qualitative and quantitative species specific-differences in tissue distribution. The gene is expressed in larval fat body but in other tissues such as gastric caeca, midgut and Malpighian tubules its expression is reduced compared to most Drosophilidae species, and in adults it is almost limited to the fat body. The comparative analysis of gene expression of two strains, which differ by a duplication, indicates that the cis elements necessary for this pattern of expression in larvae are included in the region of 1.55 kb upstream of the transcription initiation site. This new organization reveals the evolution of a different regulatory strategy to express the Adh gene in the subgenus Drosophila. PMID- 11437916 TI - Expression and function of the Drosophila melanogaster ADH in male Ceratitis capitata adults: a potential strategy for medfly genetic sexing based on gene transfer technology. AB - The aim of development of a Mediterranean fruit fly Ceratitis capitata genetic sexing strain derives from the large scale SIT programmes being carried out to control this pest. Toward this direction, we present here the male-specific expression of the Drosophila melanogaster alcohol dehydrogenase (ADH) in medfly transgenic adults generated by Minos-mediated germ line transformation. This expression pattern is obtained by using a promoter fragment of the male-specific gene MSSP-alpha2 of the medfly. We show that the heterologous enzyme is functional in the medfly oxidizing both ethanol and 2-propanol. Although leading to an approximately twofold increase of total ADH activity in male compared to female transgenic adults, these expression levels are not enough for performing genetic sexing when high doses of environmental alcohol are applied. This could be achieved either by further enhancement of the transgene expression or by generating an Adh- line to host the Minos insertions. PMID- 11437917 TI - Effect of baculovirus infection on the mRNA and protein levels of the Spodoptera frugiperda eukaryotic initiation factor 4E. AB - The cDNA sequence of eukaryotic translation initiation factor eIF4E was derived from a Spodoptera frugiperda cDNA library. Eight tryptophan residues, typical for eIF4E, are strictly conserved in the encoded 210 amino acid protein. A polyclonal antiserum detected a 26 kDa protein in lepidopteran cell lines, but not in dipteran cells. Sf21 cells have a single eIF4E gene copy, which is transcribed into a 1500 nt transcript. Infection with AcMNPV resulted in a decrease in eIF4E mRNA starting between 12 and 24 h postinfection (p.i.), while reduced eIF4E protein levels were observed at 48 h p.i. Two forms of eIF4E were recognized that differed in their iso-electric point, of which the relative abundance did not change during infection. Mutagenesis experiments using recombinant baculoviruses revealed that the variation in mobility between these two forms did not result from a difference in the phosphorylation state of Ser-202, the serine residue that corresponds with the eIF4E phosphorylation site in mammalian eIF4E. PMID- 11437918 TI - Sindbis virus-induced silencing of dengue viruses in mosquitoes. AB - Aedes aegypti were injected intrathoracically with double subgenomic Sindbis (dsSIN) viruses with inserted sequences derived from the genome of one or more of the four dengue (DEN) virus serotypes. Mosquitoes were highly resistant to challenge with homologous DEN viruses from which the effector sequences were derived, and resistance to DEN viruses was independent of the orientation of the effector RNA. dsSIN viruses designed to express RNA derived from the premembrane coding region of DEN-2 prevented the accumulation of DEN2 RNA, and C6/36 cells were highly resistant to DEN-2 virus when challenged at 2, 5 or 8 days after the initial dsSIN virus infections, even though the dsSIN-derived RNA had sharply declined at the later time points. Initiation of resistance occurred prior to or within the first 8 h after challenge with DEN-2 virus. We conclude that DEN viruses are inhibited by a mechanism similar to post-transcriptional gene silencing (PTGS) or RNA interference (RNAi) phenomena described in plants and invertebrates, respectively. The potential occurrence of PTGS or RNAi in mosquitoes and mosquito cells suggests new ways of inhibiting the replication of arthropod-borne viruses in mosquito vectors, studying vector-virus interactions, and silencing endogenous mosquito genes. PMID- 11437919 TI - High amplification of a densovirus-derived vector in larval and adult tissues of Drosophila. AB - The Lepidopteran densovirus-derived vector, pJlacZDeltaNS3, is a defective virus genome with an insertion of lacZ DNA in the viral structural protein coding sequence, and a deletion of the sequence coding the non-structural polypeptide NS3. pJlacZDeltaNS3 was injected into Drosophila eggs and the maintenance of the viral genome was monitored by expression of beta-galactosidase and by Southern blot hybridizations. Intense beta-galactosidase activity was observed in many somatic tissues of third-instar larvae and adult flies, in more than 60% of the injected animals. DNA analyses showed that staining in adult tissues correlated with the amplification of the vector. Together, these results suggest the occurrence of early events of integration of the vector into the Drosophila host genome. PMID- 11437920 TI - Phylogeny of raspberry beetles and other Byturidae (Coleoptera). AB - Nuclear ribosomal ITS2 and mitochondrial cytochrome oxidase DNA sequences were analysed from the Byturidae (Coleoptera), which includes the raspberry beetles. The secondary structure of ITS2 was plotted and interspecific changes analysed. Evidence for selection on simple sequence repeats within the ITS2 was found. Phylogenetic trees based on the mitochondrial and ribosomal sequences were compared. They were in parity, indicating they reflect the true evolutionary histories of these insects. There was no evidence for hybridization in the populations surveyed, but there was evidence that the American raspberry beetle, Byturus unicolor, is divided into at least three distinct groupings. Despite sharing a related host, the raspberry pests from America were not the most related to the European raspberry beetle. Instead, links between Byturus affinis from Japan and the American raspberry beetle suggest that this lineage originated in Asia and colonized the Western USA. PMID- 11437921 TI - Increased glycodelin levels in gynecological malignancies. AB - Glycodelin, an immunosuppressive protein with contraceptive properties, is synthesized by a variety of tissues and cell types. The ability of reproductive tissues to synthesize glycodelin is of major interest in pregnancy and disease conditions. We studied glycodelin levels in subjects with malignant gynecological tumors and in control subjects. Using a polyclonal glycodelin antibody against the synthetic glycodelin peptide sequence, an enzyme-linked immunosorbent assay (ELISA) was devised to measure plasma glycodelin levels. The assay detected as much as 5 ng/ml of glycodelin. There was a significant increase in plasma glycodelin levels in endometrial > ovarian > cervical cancer subjects when compared to those of controls. Strong expression of mRNA and protein were found in the ovarian and endometrial tumor tissues. Given glycodelin's immunosuppressive abilities, increased level of glycodelin may facilitate tumor growth in gynecological malignancies. PMID- 11437922 TI - A nerve-sparing radical hysterectomy: guidelines and feasibility in Western patients. AB - Surgical damage to the pelvic autonomic nerves during radical hysterectomy is thought to be responsible for considerable morbidity, i.e., impaired bladder function, defecation problems, and sexual dysfunction. Previous anatomical studies and detailed study of surgical techniques in various Japanese oncology centers demonstrated that the anatomy of the pelvic autonomic nerve plexus permits a systematic surgical approach to preserve these nerves during radical hysterectomy without compromising radicality. We introduced elements of the Japanese nerve-preserving techniques and carried out a feasibility study in ten consecutive Dutch patients. The technique involved three steps: first, the identification and preservation of the hypogastric nerve in a loose tissue sheath underneath the ureter and lateral to the sacro-uterine ligaments; second, the inferior hypogastric plexus in the parametrium is lateralized and avoided during parametrial transsection; third, the most distal part of the inferior hypogastric plexus is preserved during the dissection of the posterior part of the vesico uterine ligament. The clinical study showed that the procedure is feasible and safe, except possibly when used with very obese patients and patients with broad, bulky tumors. Surgical preservation of the pelvic autonomic nerves in radical hysterectomy deserves consideration in the quest to improve both cure and quality of life in cervical cancer patients. PMID- 11437923 TI - Biologic factors and response to radiotherapy in carcinoma of the cervix. AB - Ionizing radiation has been used to treat cancers for a century. However, radioresistance remains a major problem in the clinic. Recent advances in the understanding of the molecular events that occur following ionizing radiation leading to DNA damage and repair, apoptosis, and cell cycle arrests suggest new ways in which the radiation response might be manipulated. Seventy-eight cases of carcinoma of the cervix of the same stage (II A and B) were analyzed retrospectively. All patients were treated with radiotherapy (RT) with a dose varying from 35 Gy to 50 Gy with 200 cGy per fraction. Subsequent to the completion of radiotherapy, all patients underwent surgery 4-6 weeks later. On histological examination of the surgical specimens, 51% of the cases (40) showed a complete response to therapy with no viable tumor cells. 49% of cases (38) had residual tumors ranging from a small focus to lesions extending through more than half the thickness of the cervical wall. p53 (mutant), bcl-2, p21 and bax proteins were studied on the paraffin sections of the biopsies (pretreatment) of those patients who failed to respond to RT and compared to similar studies on biopsies of patients who had a complete response to RT. In addition, the minichromosome maintenance (MCM) 2 proliferative marker was also done on all cases. Expression of all proteins was done using immunohistochemsitry. In the radioresistant cases, 15% (six cases) showed positivity for bcl-2 and p21, respectively, and 34% (13 cases) showed mutant p53. None of the radiosensitive tumors were positive for the above proteins. 75% of the radiosensitive tumors (30 cases) were positive for the bax antibody, whereas 81% of the radioresistant tumors (31 cases) were negative for bax. The MCM2 proliferative marker was positive in > 80% of cells in 81.5% of radioresistant tumors (31 cases) as compared to < 40% of cells that were positive in 70% of radiosensitive tumors (28 cases). The P-value for the biological markers was calculated using the chi squared test, and was highly significant (P < 0.01) for all the parameters tested. However, there was no statistical significance by univariate analysis when the dose of radiation was analyzed with respect to the markers and the histological response. There was also no correlation between the radiation response and timing of surgery. The above data strongly suggest that bax, along with proliferative markers, could play a role in determining which tumors are likely to respond to radiation therapy. The presence of bcl-2, p21 and p53 could also be related to radioresistance of the tumors. PMID- 11437924 TI - Invasive cervical cancer and human immunodeficiency virus (HIV) infection: a South African perspective. AB - The aim of this study was to compare the prevalence and presentation of cervical cancer in HIV-positive and HIV-negative women in our local population. Six hundred and seventy-two patients with cervical cancer presented to the gynecology oncology unit of King Edward VIII Hospital, South Africa. The HIV seroprevalence among these patients was 21%. There was an increase in the background prevalence of HIV infection (1.6-32.5%) as well as a quadrupling in the prevalence of HIV infection among patients with invasive cervical cancer (5-21%) over a 10-year period. The mean ages of the HIV-negative patients and HIV-positive patients were 55.2 and 39.8 years, respectively. Most of the HIV-positive patients were in the 30- to 40-year-old age group (51%), whereas the majority of patients who were HIV negative were in the 50- to 60-year-old age group (36%). The majority of patients, irrespective of HIV status, were more likely to have late stage disease than early stage disease. There was an increase in HIV infection in patients with both types of background prevalence and among patients with invasive cervical cancer. The mean age of HIV-positive patients was 15 years younger than that of the HIV-negative patients. The majority of patients, irrespective of HIV status, presented with late stage disease. PMID- 11437925 TI - Distribution of HPV 53, HPV 73 and CP8304 in genital epithelial lesions with different grades of dysplasia. AB - To characterize the risk of malignant progression of cervical epithelial lesions associated with human papillomavirus (HPV) types of yet unknown oncogenic potential the prevalences of these HPVs in different cervical epithelial lesions of 809 patients were determined. HPV types 53, 73, and CP8304 were detected in genital specimens of 16, 22, and 12 of the patients, respectively. The ratio of prevalence in high grade dysplastic lesions or cancers and low grade dysplastic lesions or normal specimens was calculated and compared to corresponding values of well known high-risk (HR) and low-risk (LR) HPVs. For HPV 6, 11, 16, 18, 35, and 73 a ratio of 0.1, 0.2, 5.9, 6.5, 2.5, and 2.4, respectively, was calculated. The ratios of HPV53 and CP8304 were less than 1. Moreover, in contrast to HPV73, these viruses have never been detected in cancer specimens. Thus, HPV53 and CP8304 infections are probably not associated with a high risk of carcinogenesis, while HPV73 could be another HR-HPV type. PMID- 11437926 TI - A comparison of complementary and alternative medicine use by gynecology and gynecologic oncology patients. AB - Our objective was to describe and compare the use of complementary and alternative medicine (CAM) in gynecology and gynecological oncology patients. Five hundred and twenty-nine gynecology and gynecological oncology patients completed a questionnaire regarding CAM use. Overall, 56.3% of gynecology and gynecological oncology patients reported current use of CAM. Therapies used included nutritional supplements (20%), prayer as medical therapy (17%), exercise as medical therapy (12%), megavitamins (10%), and green tea (10%). While 69.5% believed CAM to be beneficial, only 31.6% discussed these therapies with their physician. The women spent a mean of $656.22 on CAM (range $0-$7,000), with 31.7% receiving some insurance reimbursement. Gynecologic oncology patients (n = 161) used CAM significantly more than gynecology patients (n = 368) (66% vs. 52%, 95% CI = 0.046-0.230, P = 0.004). Gynecological oncology patients also spent more for CAM, with a mean expenditure of $711 versus $622 by gynecology patients. Within the gynecological oncology patient group, there were 69 patients currently receiving modern medical treatments for cancer; among these patients, 58% reported using CAM; of these, 39.3% communicated their use of CAM to their physician. Patients in this group spent an average of $1,178 on CAM during their illness, with only 6.3% receiving insurance reimbursement. Benefits from CAM were perceived by 54.5% in this group. We concluded that cancer patients have a higher usage rate and expenditure for CAM, particularly while they are receiving medical therapy, and are more likely to discuss the use of alternative therapies with their physicians. CAM was perceived as helpful by patients despite the lack of scientific data about its effect. PMID- 11437927 TI - Type III radical hysterectomy after induction chemotherapy for patients with locally advanced cervical carcinoma. AB - Neoadjuvant chemotherapy followed by surgery is a promising approach in locally advanced cervical carcinoma. The aim of this study was to evaluate the feasibility, technical aspects, and clinical results of surgery after induction chemotherapy in this patient population. Forty-one untreated cervical carcinoma patients staged as IB2 to IIIB received three 21-day courses of cisplatin 100mg/m2 on day 1 and gemcitabine 1000 mg/m2 on days 1 and 8 followed by surgery or concomitant chemoradiation. The response to chemotherapy, operability, surgical/pathological findings, disease-free period, and survival of the surgically treated patients were evaluated. All 41 patients were evaluated for toxicity and 40 were evaluated for response. The overall objective response rate was 95% (95% confidence interval 88%-100%), and was complete in three patients (7.5%) and partial in 35 (87.5%). Granulocytopenia grades 3/4 occurred in 13.8% and 3.4% of the courses, respectively, whereas nonhematological toxicity was mild. Twenty-three patients underwent type III radical hysterectomy. Mean duration of surgery was 3.8 h (range 2:30-5:20), median estimated blood loss was 670 ml and median hospital stay was 5.2 days. Intraoperative complications occurred in one case (venous injury). In all but one case the resection margins were negative. Four patients (17%) had positive nodes (one node each); six (26%) had complete pathologic response, three (13%) had microscopic; and 14 (60%) macroscopic residual disease. At 24 months of maximum follow-up (median 20), the disease-free and overall survival rates were 59% and 91%, respectively. Induction chemotherapy with cisplatin/gemcitabine produced a high response rate and did not increase the difficulty of surgery. Operating time, blood loss, intraoperative complications, and hospital stay were all within the range observed for type III hysterectomy in early stage patients. We therefore conclude that type III radical hysterectomy is feasible in locally advanced cervical cancer patients who respond to chemotherapy. PMID- 11437928 TI - Microsatellite instability in uterine sarcomas. AB - Studies have shown a 15-30% frequency of microsatellite instability in endometrial cancer. In addition, we found a 21% frequency of microsatellite instability in endometrial cancer. Our aim was to investigate the presence of microsatellite instability and loss of heterozygosity in uterine sarcomas. The records of 69 women referred to Kalafong Academic and Pretoria Academic Hospital with a primary diagnosis of uterine sarcoma were reviewed. At histological review of 43 cases with a primary diagnosis of leiomyosarcoma, diagnosis of mitotically active leiomyoma was made in 21. Diagnosis of carcinosarcoma was made in 21 cases and endometrial stromal sarcoma in five. In all cases, genomic DNA was extracted from normal myometrium and tumor and analyzed for microsatellite instability and loss of heterozygosity. High-frequency microsatellite instability was absent in leiomyosarcoma, endometrial stromal sarcoma, and mitotically active leiomyomas and was observed in 1 (5%) carcinosarcoma. Loss of heterozygosity for chromosome 11 was present in 8/48 (17%) of uterine sarcomas, equally distributed between leiomyosarcomas (4/22 = 18%) and carcinosarcomas (4/21 = 19%). There was no loss of alleles in endometrial stromal sarcoma nor mitotically active leiomyomas. In conclusion, it is suggested that tumor suppressor genes may play a role in the tumorigenesis of uterine mesenchymal cells, whereas mismatch repair genes contribute to the carcinogenesis of endometrial cancer. PMID- 11437929 TI - Simultaneous vulvar intraepithelial neoplasia and Paget's disease: report of two cases. AB - We describe a very rare association between intraepithelial, extramammary Paget's disease and human papillomavirus- (HPV) negative, keratinized type of VIN III observed in two elderly women. In both cases, morphological and immunohistochemical investigation showed two heterogeneous but intimately admixed neoplastic populations of vulvar epithelium. Atypical keratinocytes stained markedly and diffusely positive for high molecular weight cytokeratins, and moderately for p53 protein and c-erbB-2 immunostainings. Paget cells were diffusely positive for CEA, EMA, and low molecular weight cytokeratins, moderately and focally for c-erbB-2 and (in one case) for S-100. Morphological and immunohistochemical phenotypic differences between Paget cells and atypical keratinocytes suggest a simultaneous and incidental association of two distinct neoplastic disorders more than a mixed carcinoma in situ of vulvar epithelium. PMID- 11437930 TI - Granulosa cell tumors of the ovary: do they have any unique ultrasonographic and color Doppler flow features? AB - The aim of this study was to describe the ultrasonographic and Doppler flow attributes of granulosa cell tumors (GCT) of the ovary and to compare these attributes to those of epithelial tumors of the ovary. Among 13,475 gynecological patients who were scanned in our ultrasound unit between 1992 and 1996, seven patients had GCT. The final diagnosis was confirmed, postoperatively, by pathologic examination and by applying the WHO classification. The ultrasonographic findings of the GCT patients were compared to those recorded in a second group of 29 patients who had been diagnosed with epithelial tumor of the ovary. The sonographic appearance of GCT of the ovary was semi-solid and the endometrium was thick in six of the seven patients. Doppler flow studies of vessels within or in the contour of the lesions showed the resistive index (RI) to be 0.448 +/- 0.018. The epithelial tumors had a similar appearance and flow pattern except for the presence of ascites in one-third of the patients. Ultrasound scanning, including color Doppler flow, did not contribute data that could assist in differentiating between GCT and epithelial tumors of the ovary. PMID- 11437931 TI - Early development of vaginal shortening during radiation therapy for endometrial or cervical cancer. AB - Vaginal necrosis can occur following radiation therapy for gynecological malignancies. The distal vaginal mucosa has a poorer radiation tolerance than the mucosa in the upper region. We examined the extent of vaginal shortening in patients treated by intravaginal brachytherapy with or without pelvic irradiation. Maximal extension of the vaginal cylinder above the pubis was measured for each insertion. We found that the difference in mean values between insertions (2.3 vs. 1.7 cm) was highly statistically significant (P < 0.0001). Our study shows that vaginal shortening can occur during the course of intracavity and external irradiation. These alterations in vaginal anatomy can have important consequences on doses received by the distal vaginal mucosa. PMID- 11437932 TI - Pretreatment hemoglobin, platelet count, and prognosis in endometrial carcinoma. AB - We analyzed whether a low pretreatment hemoglobin level is a prognostic factor in endometrial cancer and whether it is associated with thrombocytosis. Two hundred and twelve patients with endometrial cancer treated with surgery were reviewed. Data were analyzed with Pearson's chi-squared test, Fisher's exact test in contingency tables, the Mann-Whitney U-test, the Student's t-test, and Kaplan Meier estimates. Multivariate analysis was performed with the log-rank test and the Cox proportional hazard model. Thirty-nine patients (18%) had a pretreatment hemoglobin value of < 12.0 g/dL. These 39 patients had significantly higher rates of nonendometrioid histology, high-grade tumors, myometrial invasion of > 50%, adnexal involvement, lymph-vascular space involvement, and advanced FIGO stage than patients with hemoglobin > or = 12.0 g/dL. The rate of thrombocytosis was significantly higher in patients with a low hemoglobin level (36% vs. 8%, P < 0.01). The overall 5-year survival rate of patients with low pretreatment hemoglobin was 59% compared with 89% for those with hemoglobin > or = 12 g/dL (P < 0.01). In the multivariate analysis age, thrombocytosis, nonendometrioid histology, high-grade histology, and advanced FIGO stage were significantly associated with a poor prognosis whereas adnexal involvement, lymph-vascular space involvement, low hemoglobin and myometrial invasion were not. These data indicate that low pretreatment hemoglobin is a prognostic factor in patients with endometrial cancer and that it is associated with thrombocytosis. Low hemoglobin was strongly associated with other unfavorable prognostic factors so that it was significant in the univariate but not the multivariate analysis. PMID- 11437933 TI - A unique BRCA1 mutation identified in Mongolia. AB - This is a case report of genetic assessment conducted on a family residing in the third world where two sisters have presented with early onset ovarian cancer. Protein truncation testing and DNA sequencing identified a unique mutation on exon 11 (3452delA) of the BRCA1 gene. Buccal swab testing of three siblings and three offspring showed that half of the family members carried the same mutation. Currently, genetic testing in third-world countries is conducted within research budgets, as testing is not affordable or locally available for such high-risk families. Unique mutations in the BRCA1 gene that are expressed in geographically isolated groups will be useful in genetic counseling and preventative maneuvers. The only preventive management strategy available in the third world is prophylactic surgery. PMID- 11437934 TI - Scalp metastasis from squamous cell carcinoma of the cervix. AB - We describe a 45-year-old woman with squamous cell carcinoma of the cervix stage IIB, who was initially treated with radical radiotherapy. The patient developed multiple scalp metastases 8 months following her treatment. The scalp was involved in the disease as the sole anatomic site of distant cutaneous metastasis. The scalp lesions were treated with palliative radiotherapy. A search of the literature revealed only two cases of such distant metastatic involvement of the scalp from cervical cancer. PMID- 11437935 TI - Concurrent primaries of vaginal clear cell adenocarcinoma and endometrial adenocarcinoma in a 39-year old woman with in utero diethylstilbestrol exposure. AB - Diethylstilbestrol (DES) was used widely in the late 1940s in an attempt to prevent adverse pregnancy outcomes. In 1971 the US Food and Drug Administration proscribed its use for pregnancy support secondary to its association with clear cell adenocarcinoma of the vagina. Several studies in animal models demonstrated an association with endometrial cancer among offspring following in utero DES exposure. To date, there is only one case report of endometrial cancer in women exposed to DES in utero. We present the first case, to our knowledge, of a woman exposed to DES in utero who presented with double primaries of clear cell cancer of the vagina concomitant with endometrial cancer. PMID- 11437936 TI - Primary choriocarcinoma of the vulva. AB - Primary extrauterine choriocarcinoma is very rare, found mostly in the genital tract (tube, cervix, ovary, vagina). Other sites such as lungs, gastrointestinal tract, heart, brain and submentum also have been reported. A 31-year-old woman presented with abnormal uterine bleeding 6 weeks after her last menstrual cycle. Her HCG titer level was 900 mIu/ml. She underwent dilation and curettage (D & C). Pathology failed to reveal any chorionic villi. Further evaluation was done, including a second D & C, laparoscopy, exploratory laparotomy, CT, MRI and ultrasound which all failed to define a source for the elevated HCG titer which subsequently rose to 95,000 mIu/ml 130 days after her last menstrual cycle. At that time a vulvar mass was observed. Fine needle aspiration (FNA) was positive for choriocarcinoma. The patient was treated with one course of methotrexate 60 mg intramuscularly every other day for 5 days. Actinomycin-D 0.5 mg given intravenously daily for 5 days was added to courses 2-7. Radiotherapy (4000 rads) was applied concomitant with the chemotherapy. The remainder of the mass was subsequently excised. The patient has remained with no evidence of disease for 10 years. This is the first case report of primary vulvar choriocarcinoma. PMID- 11437938 TI - Scarring alopecia and the dermatopathologist. AB - BACKGROUND: The evaluation of patients with cicatricial alopecia is particularly challenging, and dermatopathologists receive little training in the interpretation of scalp biopsy specimens. Accurate interpretation of specimens from patients with hair disease requires both qualitative (morphology of follicles, inflammation, fibrosis, etc.) and quantitative (size, number, follicular phase) information. Much of this data can only be obtained from transverse sections. In most cases, good clinical/pathologic correlation is required, and so clinicians should be expected to provide demographic information as well as a brief description of the pattern of hair loss and a clinical differential diagnosis. RESULTS: The criteria used to classify the various forms of cicatricial alopecia are relatively imprecise, and so classification is controversial and in a state of evolution. There are five fairly distinctive forms of cicatricial alopecia: 1) chronic, cutaneous lupus erythematosus (discoid LE); 2) lichen planopilaris; 3) dissecting cellulitis (perifolliculitis abscedens et suffodiens); 4) acne keloidalis; and 5) central, centrifugal scarring alopecia (follicular degeneration syndrome, folliculitis decalvans, pseudopelade). Not all patients with cicatricial alopecia can be confidently assigned to one of these five entities, and "cicatricial alopecia, unclassified" would be an appropriate label for such cases. CONCLUSION: The histologic features of five forms of cicatricial alopecia are reviewed. Dermatopathologists can utilize a "checklist" to catalog the diagnostic features of scalp biopsy specimens. In many, but not all, cases the information thus acquired will "match" the clinical and histologic characteristics of a form of cicatricial alopecia. However, because of histologic and clinical overlap between the forms of cicatricial alopecia, a definitive diagnosis cannot always be rendered. PMID- 11437939 TI - Comprehensive analysis of 112 melanocytic skin lesions demonstrates microsatellite instability in melanomas and dysplastic nevi, but not in benign nevi. AB - INTRODUCTION: the length of DNA repetitive sequences (microsatellite instability (MSI)) represent distinct tumorigenic pathways associated with several familial and sporadic tumors. MATERIAL AND METHODS: To investigate the prevalence and frequency of MSI in melanocytic lesions, the polymerase chain reaction (PCR) based microsatellite assay was used to examine formalin-fixed, paraffin-embedded tissues of 30 benign melanocytic nevi, 60 melanocytic dysplastic nevi (MDN), and 22 primary vertical growth phase cutaneous malignant melanomas (CMM). Twenty-four microsatellite markers at the 1p, 2p, 3p, 4q and 9p chromosomal regions were used. RESULTS: MSI was found at 1p and 9p in MDN and CMM but not in benign melanocytic nevi. The overall prevalence of MSI was 17/60 (28%) in MDN and 7/22 (31%) in CMM. The frequency of MSI ranged from 2/24 (9%) to 4/24 (17%) and was most commonly found at D9S162. There was a statistically significant correlation between degree of atypia and frequency of MSI (p<0.001) in MDN. There were two MSI banding patterns: band shifts and additional bands. CONCLUSIONS: The data presented revealed the presence of low-frequency MSI (MSI-L) at the 1p and 9p regions in both MDN and CMM. Whether the MSI-L pattern reflects a defect in mismatch repair genes is still to be determined. PMID- 11437940 TI - Heteroduplex analysis of T-cell receptor gamma gene rearrangement as an adjuvant diagnostic tool in skin biopsies for erythroderma. AB - BACKGROUND: Erythroderma, defined as red skin covering most of the body surface often accompanied or followed by exfoliation, is the clinical manifestation of at least six different underlying etiologies with allergic or irritant contact dermatitis, atopic/asteotic dermatitis, pityriasis rubra pilaris (PRP), psoriasis, and seborrheic dermatitis accounting for the majority of cases. Approximately 10% of cases are due to adverse drug reactions with roughly another 10% due to cutaneous T-cell lymphoma (CTCL), predominantly mycosis fungoides, or leukemia. It is clear from multiple studies that the clinical diagnosis of the underlying entity is often difficult, as these diseases can present in a very similar fashion. A skin biopsy is usually employed in this setting as a diagnostic tool. However, the histopathologic diagnosis of the underlying cause is complicated by the subtlety of the distinguishing histologic features. In this situation, an ancillary technique demonstrating the presence of a monoclonal T cell proliferation could help to rule in or out CTCL in cases that clinically and histopathologically do not allow a definitive diagnosis. METHODS: We retrospectively studied 25 biopsies from sixteen patients who presented to the Stanford Dermatology Clinic with erythroderma. We examined the specimens morphologically and analyzed the gamma chain of the T-cell receptor (TCR- gamma) by polymerase chain reaction (PCR) followed by heteroduplex analysis for clonality. We then correlated the results of our PCR and heteroduplex analyses with the patients' clinical outcomes. RESULTS: Four biopsies, from three patients, contained clonal TCR-gamma rearrangements; the four biopsies, all of which were equivocal histologically, correlated to diagnoses of mycosis fungoides (MF) or Sezary syndrome (SS). Twenty-one biopsies contained polyclonal T-cell populations. Eighteen of these biopsies represent patients with inflammatory dermatoses. Three of these biopsies, all of which were taken from a single patient, correlate to a diagnosis of MF. CONCLUSION: TCR-gamma PCR heteroduplex analysis seems to represent an important adjuvant diagnostic tool that, used in conjunction with histopathology and clinical history, could help to clarify the underlying etiology of erythroderma. PMID- 11437941 TI - CD45RA+ immunophenotype in mycosis fungoides: clinical, histological and immunophenotypical features in 22 patients. AB - BACKGROUND: Mycosis fungoides (MF) is a cutaneous T-cell lymphoma (CTCL) usually characterized by a T-helper memory phenotype (CD3+, CD4+, CD8-, CD45R0+). Aberrant phenotypes are more commonly seen in the tumor stages. CD45RA expression has so far been documented in only a few cases of CD8+ or TCR gamma delta+ CTCL and in some pagetoid reticulosis cases. METHODS: Two hundred and fifteen MF patients were immunophenotyped in our laboratory between January 1992 and June 2000 and 22 cases of CD45RA+ MF (8.7%) were identified by immunohistochemical analysis. RESULTS: The majority of these CD45RA+ patients (20/22) showed a patch plaque stage disease and an indolent clinical course, as expected in early-stage MF. The remaining 2 patients presented with stage IIB and IVA MF, and were characterized by an aggressive clinical course, with systemic spread. The immunohistochemical analysis revealed that CD45RA+ neoplastic cells belonged to the memory compartment, displaying a CD62L-, CD11a+, CD29+ phenotype. Most patients showed aberrant phenotypes, with a loss of T-cell lineage markers and expression of cytotoxic molecules or gamma-delta chain of the T-cell receptor. CONCLUSIONS: Our data show that CD45RA+ MF is a rare variant of CTCL and shares with the classic MF cases both the clinical features and disease course, even if it is characterized by a higher incidence of immunopathological abnormalities. PMID- 11437942 TI - Polymerase chain reaction (PCR) for human herpesvirus 8 and heteroduplex PCR for clonality assessment in angiolymphoid hyperplasia with eosinophilia and Kimura's disease. AB - BACKGROUND: Recently, human herpesvirus 8 (HHV-8) has been isolated from almost all cases of Kaposi's sarcoma. It has not been found in most cutaneous hemangioproliferative disorders other than Kaposi's sarcoma. Benign vascular lesions including Kimura's disease were not found to contain the HHV-8 DNA sequence. However, there has been contradictory data concerning the presence of HHV-8 in angiolymphoid hyperplasia with eosinophilia (ALHE). Clonality studies in ALHE and Kimura's disease were rare. METHODS: We performed polymerase chain reaction (PCR)-based analysis to determine whether HHV-8 is present and heteroduplex analysis of rearranged T-cell receptor (TCR) gene for clonality assessment in paraffin-embedded skin biopsy samples of 7 ALHE and 2 Kimura's disease, taken from immunocompetent patients. RESULTS: HHV-8 could not be identified in all the cases of ALHE and Kimura's disease. Although 2 cases (2/7) of ALHE and 2 cases (2/2) of Kimura's disease showed positive result for PCR analysis of TCR, all the cases were negative for heteroduplex-PCR. CONCLUSIONS: We suggest that HHV-8 may not involve in a pathogenetic role in ALHE and Kimura's disease and the failure to demonstrate clonality may be consistent with the reactive nature of these diseases and lack of malignant transformation. In addition, heteroduplex-PCR can be applied to confirm doubtful cases of lymphoma in that heteroduplex-PCR is more specific than PCR as seen in our study. PMID- 11437943 TI - Subcutaneous nodules in Whipple's disease. AB - BACKGROUND: Cutaneous findings other than hyperpigmentation are rare in Whipple's disease. CASE REPORT: We present the case of a 59-year-old man previously diagnosed with Whipple's disease by duodenal biopsy, who developed red-brown, painful, subcutaneous nodules on the buttocks, thighs, arms and legs. Biopsy of these nodules showed a septal panniculitis and foamy macrophages containing PAS positive, diastase resistant intracytoplasmic material, characteristic of Whipple's disease and similar to that observed in the duodenal biopsy. Ultrastructurally, this material in the histiocytes corresponded to degenerated bacilli. CONCLUSIONS: This is the fourth documented case of subcutaneous involvement by Whipple's disease. One should consider the possibility of Whipple's disease in any patient who presents with symptoms compatible with that condition who demonstrates septal panniculitis with a large amount of foamy histiocytes. PMID- 11437944 TI - Primary cutaneous Langerhans cell histiocytosis showing malignant phenotype in an elderly woman: report of a fatal case. AB - BACKGROUND: Langerhans cell histiocytosis (LCH) is a proliferating disorder of Langerhans cells (LC) that are characterized by the presence of Birbeck granules. LCH has been considered to be a disease of childhood and there have been limited cases of adult LCH. We report here a fatal case of histiocytic tumor showing Langerhans cell phenotype, arising in the skin of a 74-year-old woman. METHOD: In addition to routine histological and immunohistological sections, electron microscopic examination and human androgen receptor gene (HUMARA) assays were performed. RESULTS: Histological examination revealed a dense dermal infiltrative proliferation of fairly large tumor cells with abundant ill-defined cytoplasms and oval or indented nuclei, in which numerous eosinophils were associated with the tumor nests. Tumor cells were positive with anti-S-100 and CD1a antibodies but negative with HMB-45 antibody or other epithelial or lymphocytic markers. Ultrastructural analysis showed typical Birbeck granules in the cytoplasm of the tumor cells. HUMARA assay of the tumor tissue revealed the nonrandom X inactivation pattern, indicating the clonal proliferation. CONCLUSIONS: We diagnosed this tumor as Langerhans cell histiocytosis with a clonal neoplastic phenotype originated in the skin. Although she demonstrated no recurrence nor metastases for 6 months after surgical resection of primary skin lesion and subsequent radiation therapy, the tumor recurred and extended multisystemically, and she died of multiple organ failure 14 months after initial diagnosis. Therefore, we would like to emphasize this case as LC "sarcoma" or "malignant" LCH. PMID- 11437945 TI - Trichoepithelioma with "monster" stromal cells. AB - Trichoepithelioma is a benign tumor of trichogenic origin which appears predominantly in childhood or in young adults. Different forms have been described according to clinical and histological features. The authors report a unique variant of trichoepithelioma arising on the limb of a 27-year-old man. The tumor was characterized by the mixture of an atypical fibroxanthomatous proliferation and basaloid epithelial strands of trichoepithelioma. Such histological features have not been previously reported. It raises the question of an additional variant of hair follicle tumor with a mixed epithelial and mesenchymal proliferation. PMID- 11437946 TI - Intrauterine epidermal necrosis. AB - BACKGROUND: Epidermal necrosis in a neonate is an uncommon event with a variety of potential cases. RESULT: We report a case of intrauterine epidermal necrosis in a preterm infant, with death occurring soon after birth. The histopathology of the denuded skin revealed full-thickness epidermal necrosis and calcification within both the epidermis and follicular structures. CONCLUSION: We believe this represents the fourth reported case of lethal intrauterine epidermal necrosis and follicular calcification. PMID- 11437947 TI - Study of the conformational profile of the cyclohexane analogs of L phenylalanine. AB - The conformational profile of the conformationally constrained cyclohexane analogs of phenylalanine (1-amino-2-phenylcyclohexanecarboxylic acids, c6Phe) was assessed using computational methods. For this purpose, the conformational space of the N-acetyl methylamide derivatives of the stereoisomers (2S,3R)c6Phe and (2S,3S)c6Phe was explored by computing their respective Ramachandran maps, and low-energy minima were characterized at molecular mechanics level by means of the AMBER program, using the parm94 force field set of parameters. In order to assess the performance of the molecular mechanics calculations, each of the low-energy conformations was also investigated further at the ab initio level. Accordingly, the molecular mechanics geometries were used as starting conformations to perform full geometry optimizations at the Hartree-Fock level, using a 6-31G(d) basis set. Analysis of the results revealed that the cyclohexane structure directly induces some restrictions on the backbone, and constrains the orientation of the aromatic side-chain to two narrow regions for each stereoisomer. The conformational profile of these amino acids is then explained on the grounds of the interaction between the rigidly held phenyl ring and the main chain NH and CO groups. The results obtained are in good accordance with the experimental observations. PMID- 11437948 TI - Functional preference of the constituent amino acid residues in a phage-library based nonphosphorylated inhibitor of the Grb2-SH2 domain. AB - A nonphosphorylated disulfide-bridged peptide, cyclo(Cys-Glu1-Leu-Tyr-Glu-Asn-Val Gly-Met-Tyr9-Cys)-amide (termed G1) has been identified, by phage library, that binds to the Grb2-SH2 domain but not the src SH2 domain. Synthetic G1 blocks the Grb2-SH2 domain association (IC50 of 15.5 microM) with natural phosphopeptide ligands. As a new structural motif that binds to the Grb2-SH2 domain in a pTyr independent manner, the binding affinity of G1 is contributed by the highly favored interactions of its structural elements interacting with the binding pocket of the protein. These interactions involve side-chains of amino acids Glu1, Tyr3, Glu4, Asn5, and Met8. Also a specific conformation is required for the cyclic peptide when bound to the protein. Ala scanning within G1 and molecular modeling analysis suggest a promising model in which G1 peptide binds in the phosphotyrosine binding site of the Grb2-SH2 domain in a beta-turn-like conformation. Replacement of Tyr3 or Asn5 with Ala abrogates the inhibitory activity of the peptide, indicating that G1 requires a Y-X-N consensus sequence similar to that found in natural pTyr-containing ligands, but without Tyr phosphorylation. Significantly, the Ala mutant of Glu1, i.e. the amino acid N terminal to Y3, remarkably reduces the binding affinity. The position of the Glu1 side-chain is confirmed to provide a complementary role for pTyr3, as demonstrated by the low micromolar inhibitory activity (IC50 = 1.02 microM) of the nonphosphorylated peptide 11, G1(Gla1), in which Glu1 was replaced by gamma carboxy-glutamic acid (Gla). PMID- 11437949 TI - Influence of hydrophobic interactions on the conformational adaptability of the beta-Ala residue. AB - The chemical synthesis and X-ray crystal structure analysis of a model peptide incorporating a conformationally flexible beta-Ala residue: Boc-beta-Ala-Pda, 1 (C23H46N2O3: molecular weight = 398.62) have been described. The peptide crystallized in the crystal system triclinic with space group P21: a = 5.116(3) A, b = 5.6770(10) A, c = 21.744(5) A; alpha = 87.45 degrees, beta = 86.87 degrees, gamma = 90.0 degrees; Z = 1. An attractive feature of the crystal molecular structure of 1 is the induction of a reasonably extended backbone conformation of the beta-Ala moiety, i.e. the torsion angles phi approximately 115 degrees, mu approximately 173 degrees and psi approximately 122 degrees, correspond to skew-, trans and skew+ conformation, respectively, by an unbranched hydrophobic alkyl chain, Pda, which prefers an all-anti orientation (theta1 approximately -153 degrees, theta2 approximately ellipsis theta14 approximately +/-178 degrees ). The observation is remarkable because, systematic conformational investigations of short linear beta-Ala peptides of the type Boc beta-Ala-Xaa-OCH3 (Xaa = Aib or Acc6) have shown that the chemical and stereochemical characters of the neighboring moieties may be critical in dictating the overall folded and/or unfolded conformational features of the beta Ala residue. The overall conformation of 1 is typical of a 'bar'. It appears convincing that, in addition to a number of hydrophobic contacts between the parallel arranged molecules, an array of conventional N-HellipsisO=C intermolecular H-bonding interactions stabilize the crystal molecular structure. Moreover, the resulting 14-membered pseudo-ring motif, generated by the amide amide interactions between the adjacent molecules, is completely devoid of nonconventional C-HellipsisO interaction. The potentials of the conformational adaptation of the beta-Ala residue, to influence and stabilize different structural characteristics have been highlighted. PMID- 11437950 TI - Synthesis, biological activity, and solution structures of a cyclic dodecapeptide from the EGF-2 domain of blood coagulation factor VII. AB - The cyclic dodecapeptide, disulfide-cyclo-[H-Cys-Val-Asn-Glu-Asn-Gly-Gly-Cys(Acm) Glu-Gln-Tyr-Cys-OH], which corresponds to the 91-102 sequence of the second epidermal growth factor domain of human blood coagulation factor VII, was synthesized using solid-phase procedures. It was shown to be an inhibitor at the key step in the induction of coagulation by the extrinsic pathway, i.e. the factor VII/tissue factor-catalyzed activation of coagulation factor X. The solution structure of this peptide was investigated by NMR spectroscopy and was computer-modeled via molecular mechanics. Structures were calculated based on 112 distance and nine dihedral angle constraints. The resulting backbone structures were classified into two structural subsets: one which exhibited a twisted '8' shaped folding and another describing an open, circular 'O' outline. The local backbone structures of segments Asn3-Glu4-Asn5, Gly7-Cys8 and Gln10-Tyr11 were well preserved among the two subsets. Apart from the unrestrained N- and C termini, Gly6 and Glu9 sites exhibited marked local disorder between the two subsets, suggesting localized flexible hinges likely to govern tertiary structure interconversion between the two subsets. Two transient hydrogen bonds were identified from pH chemical shift titrations by matching the pKa values of NH and carboxylate groups, which supported the occurrence of the '8' structure, and agreed with temperature coefficients of peptidyl NH resonances. Structure function relationships of the peptide were discussed in terms of the likely physiological function of the disulfide-bonded loop in factor VII which the peptide represents. PMID- 11437951 TI - Combined use of ESI-MS and UV diode-array detection for localization of disulfide bonds in proteins: application to an alpha-L-fucosidase of pea. AB - A simplified strategy is described for the assignment of disulfide bonds in proteins of medium to high molecular mass (10-30 kDa). The method combines the use of high-performance liquid chromatography coupled to electrospray ionization mass spectrometry (HPLC-ESI-MS) and HPLC with UV diode-array detection (HPLC diode array). The denatured protein is subjected to proteolysis and the peptide mixture is divided into three fractions: (i) underivatized peptides, (ii) ethylpyridylated peptides, and (iii) reduced and ethylpyridylated peptides. The three peptide ensembles are then subjected to chromatographic and spectroscopic analysis. A systematic methodology is described to analyze the large amount of data obtained. The method was applied to the localization of disulfide bonds in alpha-L-fucosidase from pea. The two disulfide bonds were located between residues Cys64 and Cys109 and between Cys162 and Cys169, while Cys127 was free. PMID- 11437952 TI - Mass spectrometric evaluation of synthetic peptides as primary structure models for peptide and protein deamidation. AB - Solid-phase peptide synthesis and deamidation measurements using a novel mass spectrometric technique were carried out for 94 model asparaginyl peptides from 3 to 13 residues in length. Deamidation rates of these peptides in pH 7.4, 37.0 degrees C, 0.15 M Tris-HCl buffer were measured and evaluated. It was found that they validate the use of pentapeptide models as surrogates for the primary sequence dependence of peptide and protein deamidation rates and the discovery by difference of secondary, tertiary and quaternary structure effects. Deamidation of the pentapeptide models, compared with that of longer peptides of more intricate structure, is discussed, and the application of this technique to deamidation measurement of intact proteins is demonstrated. PMID- 11437953 TI - Tritiated photoactivatable analogs of the native human thrombin receptor (PAR-1) agonist peptide, SFLLRN-NH2. AB - Six photoactivatable analogs of the human thrombin receptor activating peptide (TRAP), SFLLRN-NH2, were synthesized by substituting the photoactive amino acid, p-benzoylphenylalanine (Bpa), into each position of the peptide sequence. Platelet aggregation assays indicated that the peptides with Bpa substitutions at positions 3 to 6 retained agonist activity. These peptides were prepared in tritiated form as potential thrombin receptor photoaffinity labels. The [3H]Bpa containing analogs were constructed by resynthesizing the peptides with the amino acid, 4-benzoyl-2',5'-dibromophenylalanine (Br2Bpa), and subjecting the purified peptides to Pd-catalyzed tritiodebromination. The radiochemical yields for the reductive tritiation were < 2% for peptides with [3H]Bpa in the third and fourth positions, and between 7 and 16% for the peptides with substitutions at the fifth and sixth positions. The low yields were due to over-reduction of the Bpa carbonyl group and nonspecific degradation during reductive tritiation. This report describes the first use of Br2Bpa for the preparation of tritiated photoactivatable peptides. PMID- 11437954 TI - Phage randomization in a charybdotoxin scaffold leads to CD4-mimetic recognition motifs that bind HIV-1 envelope through non-aromatic sequences. AB - Binding of HIV-1 gp120 to T-cell receptor CD4 initiates conformational changes in the viral envelope that trigger viral entry into host cells. Phage epitope randomization of a beta-turn loop of a charybdotoxin-based miniprotein scaffold was used to identify peptides that can bind gp120 and block the gp120-CD4 interaction. We describe here the display of the charybdotoxin scaffold on the filamentous phage fUSE5, its use to construct a beta-turn library, and miniprotein sequences identified through library panning with immobilized Env gp120. Competition enzyme-linked immunosorbent assay (ELISA) identified high frequency phage selectants for which specific gp120 binding was competed by sCD4. Several of these selectants contain hydrophobic residues in place of the Phe that occurs in the gp120-binding beta-turns of both CD4 and previously identified scorpion toxin CD4 mimetics. One of these selectants, denoted TXM[24GQTL27], contains GQTL in place of the CD4 beta-turn sequence 40QGSF43. TXM[24GQTL27] peptide was prepared using solid-phase chemical synthesis, its binding to gp120 demonstrated by optical biosensor kinetics analysis and its affinity for the CD4 binding site of gp120 confirmed by competition ELISA. The results demonstrate that aromatic-less loop-containing CD4 recognition mimetics can be formed with detectable envelope protein binding within a beta-turn of the charybdotoxin miniprotein scaffold. The results of this work establish a methodology for phage display of a charybdotoxin miniprotein scaffold and point to the potential value of phage-based epitope randomization of this miniprotein for identifying novel CD4 mimetics. The latter are potentially useful in deconvoluting structural determinants of CD4-HIV envelope recognition and possibly in designing antagonists of viral entry. PMID- 11437955 TI - Optimized aminolysis conditions for cleavage of N-protected hydrophobic peptides from solid-phase resins. AB - Solid-phase synthesis and aminolysis cleavage conditions were optimized to obtain N- and C-terminally protected hydrophobic peptides with both high quality and yield. Uncharged 'WALP' peptides, consisting of a central (Leu-Ala)n repeating unit (where n = 5, 10.5 or 11.5) flanked on both sides by Trp 'anchors', and gramicidin A (gA) were synthesized using 9-fluorenylmethoxycarbonyl chemistry from either Wang or Merrifield resins. For WALP peptides, the N-terminal amino acid was capped by coupling N-acetyl- or N-formyl-Ala or -Gly to the peptide/resin or by formylation of the completed peptide/resin with para nitrophenylformate (p-NPF). N-Terminal acetyl- or formyl-Ala racemized when coupled as an HOBt-ester to the resin-bound peptide, but not when the peptide was formylated with p-NPF. Racemization was avoided at the last step by completing the peptide with acetyl- or formyl-Gly. For both WALP peptides and gA, cleavage conditions using ethanolamine or ethylenediamine were optimized as functions of solvent, time, temperature and resin type. For WALP peptides, maximum yields of highly pure peptide were obtained by cleavage with 20% ethanolamine or ethylenediamine in 80% dichloromethane for 48 h at 24 degrees C. N-Acetyl protected WALP peptides consistently gave higher yields than those protected with N-formyl. For gA, cleavage with 20% ethanolamine or ethylenediamine in 80% dimethylformamide for 48 h at 24 degrees C gave excellent results. For both WALP peptides and gA, decreasing the cleavage time to 4 h and increasing the temperature to 40-55 degrees C resulted in significantly lower yields. The inclusion of hexafluoroisopropanol in the cleavage solvent mixture did not improve yields for either gA or WALP peptides. PMID- 11437956 TI - Stability and CTL activity of N-terminal glutamic acid containing peptides. AB - Several cytotoxic T lymphocyte peptide-based vaccines against hepatitis B, human immunodeficiency virus and melanoma were recently studied in clinical trials. One interesting melanoma vaccine candidate alone or in combination with other tumor antigens, is the decapeptide ELA. This peptide is a Melan-A/MART-1 antigen immunodominant peptide analog, with an N-terminal glutamic acid. It has been reported that the amino group and gamma-carboxylic group of glutamic acids, as well as the amino group and gamma-carboxamide group of glutamines, condense easily to form pyroglutamic derivatives. To overcome this stability problem, several peptides of pharmaceutical interest have been developed with a pyroglutamic acid instead of N-terminal glutamine or glutamic acid, without loss of pharmacological properties. Unfortunately compared with ELA, the pyroglutamic acid derivative (PyrELA) and also the N-terminal acetyl-capped derivative (AcELA) failed to elicit cytotoxic T lymphocyte (CTL) activity. Despite the apparent minor modifications introduced in PyrELA and AcELA, these two derivatives probably have lower affinity than ELA for the specific class I major histocompatibility complex. Consequently, in order to conserve full activity of ELA, the formation of PyrELA must be avoided. Furthermore, this stability problem is worse in the case of clinical grade ELA, produced as an acetate salt, like most of the pharmaceutical grade peptides. We report here that the hydrochloride salt, shows higher stability than the acetate salt and may be suitable for use in man. Similar stability data were also obtained for MAGE-3, another N-terminal glutamic acid containing CTL peptide in clinical development, leading us to suggest that all N-terminal glutamic acid and probably glutamine-containing CTL peptide epitopes may be stabilized as hydrochloride salts. PMID- 11437957 TI - Studies on analogs of a peptide derived from alpha-fetoprotein having antigrowth properties. AB - A 34-amino acid portion of the third domain of alpha-fetoprotein possesses antigrowth and anticancer activities. Three analogs of this sequence were chemically synthesized, in which the two cysteines of the original sequence were replaced by alanines, glycines or serines. The original cysteine and alanine peptides formed trimers at 0.20 g/L in pH 7.4 phosphate buffer, and the glycine and serine peptides formed dimers. Trimer preparations were more potent in inhibiting estrogen-induced growth in the mouse uterine assays than the two dimeric oligomers. Of salient importance is that the alanine peptide retained its trimeric form in solution much longer than the cysteine peptide. Antigrowth assays were performed starting with stock solutions at a peptide concentration of 0.20 g/L, because at very high peptide concentration (8.0 g/L) the peptides aggregated extensively. All the peptides, although differing in biological activity, had almost identical secondary structures. Unlike alpha-fetoprotein, the three peptides have low amounts of alpha-helix. Trifluoroethanol has the ability to convert peptides into a helical conformation when they have a propensity for that structure. At trifluoroethanol concentrations of 20% and higher, the alanine and glycine peptides were changed into highly helical structures. PMID- 11437959 TI - Students and student life. PMID- 11437960 TI - More on medical student stress. PMID- 11437961 TI - Medical student debt in the United Kingdom. PMID- 11437962 TI - Alcohol and medical students. PMID- 11437963 TI - The reliability of logbook data of medical students: an estimation of interobserver agreement, sensitivity and specificity. AB - OBJECTIVE: Logbooks are widely used in medical schools as an evaluation tool to assess students' progress towards objectives. To estimate whether students fill in their logbooks reliably, we measured interobserver agreement by comparing doctors' data and students' data. METHOD: Completed logbooks were collected at two subdivisions of the department of Internal Medicine at the University Hospital of Groningen. The logbook contains 231 preprinted diseases. Doctors and students recorded the diseases they had encountered. Interobserver agreement, expressed by the Jaccard coefficient (J), was calculated for the complete set of diseases and for a subset of core diseases. To assess the kinds of errors which students made, sensitivity and specificity were determined. RESULTS: Logbook data of doctors and students are not fully consistent (mean J for the complete set of diseases was.23 and for the core diseases.36). The quality of the logbook data is high in the sense that students do not record many false identifications (mean specificity for the complete set of diseases and for the core diseases were.96 and.93, respectively); the quality is poor in the sense that students do not record all the diseases which could be seen at the department (mean sensitivity for the complete set of diseases is.36 and for the core diseases it is.51). CONCLUSION: This study shows inconsistencies in recording diseases in a logbook by students compared with doctors. In particular the diseases which are present at a department are under-reported by students. Supervision and feedback are important mechanisms to optimize the students' use of (1) all diseases which could be encountered and (2) the logbook. PMID- 11437964 TI - Student feedback in problem based learning: a survey of 103 final year students across five Ontario medical schools. AB - CONTEXT: Problem based learning (PBL) has become an integral component of medical curricula around the world. In Ontario, Canada, PBL has been implemented in all five Ontario medical schools for several years. Although proper and timely feedback is an essential component of medical education, the types of feedback that students receive in PBL have not been systematically investigated. OBJECTIVES: In the first multischool study of PBL in Canada, we sought to determine the types of feedback (grades, written comments, group feedback from tutor, individual feedback from tutor, peer feedback, self-assessment, no feedback) that students receive as well as their satisfaction with these different feedback modalities. SUBJECTS AND METHODS: We surveyed a sample of 103 final year medical students at the five Ontario schools (University of Toronto, McMaster University, Queens University, University of Ottawa and University of Western Ontario). Subjects were recruited via E-mail and were asked to fill out a questionnaire. RESULTS: Many students felt that the most helpful type of feedback in PBL was individual feedback from the tutor, and indeed, individual feedback was one of the more common types of feedback provided. However, although students also indicated a strong preference for peer and group feedback, these forms of feedback were not widely reported. There were significant differences between schools in the use of grades, written comments, self-assessment and peer feedback, as well as the immediacy of the feedback given. CONCLUSIONS: Across Ontario, students do receive frequent feedback in PBL. However, significant differences exist in the types of feedback students receive, as well as the timing. Although rated highly by students at all schools, the use of peer feedback and self-assessment is limited at most, but not all, medical schools. PMID- 11437965 TI - Medical student interviewing skills and mother-reported satisfaction and recall. AB - BACKGROUND: Providing feedback to medical students about their interviewing skills is an important component of teaching programmes. There is very little information about mothers' views of medical student consultations in paediatrics, and in particular about what mothers consider to be the key elements of a successful consultation. Patient-centred interviewing is a model which emphasizes the active seeking of patient views. In association with appropriate clinical skills, it is reported to promote improved health outcomes. OBJECTIVES: To examine whether greater medical student clinical competence and more frequent use of patient-centred techniques is associated with higher maternal satisfaction, higher maternal rating of the medical student's interpersonal skills, and greater maternal recall of relevant diagnosis and treatment recommendations. METHOD: Two standardized 'medical student' videotaped interviews were created based on actual senior medical student consultations. Interview A demonstrated both higher student clinical competence and higher patient-centredness compared with interview B. Both videotaped interviews were viewed and then rated, using a questionnaire, by 11 mothers attending a teaching general practice. RESULTS: Significantly higher mean scores, indicating greater maternal satisfaction, were associated with interview A (P < 0.01 for all measures). Accurate recall for diagnosis and management was also significantly greater after interview A (mean diagnosis recall, interview A 35%, interview B 14%, P < 0.01; mean management recall, interview A 95%, interview B 57%, P < 0.01). CONCLUSIONS: Maternal satisfaction and recall were higher following a more clinically competent and patient-centred medical student interview. Maternal ratings of student interviews could be used as an additional method of assessment as well as providing feedback to medical students on their interview skills development. PMID- 11437966 TI - Learning to consult with computers. AB - OBJECTIVE: To develop and evaluate a strategy to teach skills and issues associated with computers in the consultation. INTERVENTION: An overview lecture plus a workshop before and a workshop after practice placements, during the 10 week general practice (GP) term in the 5th year of the University of Melbourne medical course. DESIGN: Pre- and post-intervention study using a mix of qualitative and quantitative methods within a strategic evaluation framework. OUTCOME MEASURES: Self-reported attitudes and skills with clinical applications before, during and after the intervention. RESULTS: Most students had significant general computer experience but little in the medical area. They found the workshops relevant, interesting and easy to follow. The role-play approach facilitated students' learning of relevant communication and consulting skills and an appreciation of issues associated with using the information technology tools in simulated clinical situations to augment and complement their consulting skills. The workshops and exposure to GP systems were associated with an increase in the use of clinical software, more realistic expectations of existing clinical and medical record software and an understanding of the barriers to the use of computers in the consultation. CONCLUSIONS: The educational intervention assisted students to develop and express an understanding of the importance of consulting and communication skills in teaching and learning about medical informatics tools, hardware and software design, workplace issues and the impact of clinical computer systems on the consultation and patient care. PMID- 11437967 TI - Students' perception of medical school stress and their evaluation of a wellness elective. AB - CONTEXT: Because medical education is known to be demanding and stressful, Case Western Reserve University developed a programme which included a wellness elective which focused on stress reduction and personal wellness. OBJECTIVES: The purpose of this study was to explore students' perceptions of medical school stress and to assess their perspective on the wellness elective. SUBJECTS: The essays of 60 medical students enrolled in the wellness elective were used. METHODS: The essays were analysed using qualitative methodology. Results were validated by questionnaires mailed to the students 2 years later. RESULTS: It was found that the students felt that: (1) wellness issues should be important for physicians; (2) their own wellbeing had been diminished by the burden of information to be learned in medical school; (3) talking to peers was a useful coping mechanism, and (4) the elective gave permission to engage in wellness activities without additional guilt. CONCLUSIONS: Based on the students' responses, a wellness elective could be a useful addition to the first- or second year medical curriculum. PMID- 11437968 TI - Medical student and faculty perceptions of desirable primary care teaching site characteristics. AB - BACKGROUND: As the need to expand and improve primary care teaching experiences has mushroomed, the need to identify desirable preceptor and site characteristics has also grown. PURPOSE: The current study was designed to assess the relative importance students and preceptors place on site versus preceptor characteristics. METHODS: After a required year-long primary care experience, third-year medical students (n=39) and primary care preceptors (n=20) completed a Q-sort exercise. RESULTS: In all analyses preceptor characteristics were ranked highest. The highest ranked item for students and preceptors was 'The preceptor allows student to assume increasing levels of responsibility'. Seven of the highest ranked items appear on both groups' top 10 list. CONCLUSIONS: There is a high level of agreement between students and preceptors that preceptor rather than site characteristics make the vital difference in primary care educational experiences. When students and preceptors disagree, it is because preceptors see their role- modelling characteristics related to relationships with patients as important, while students are more interested in the hands-on learning opportunities afforded by the preceptor. PMID- 11437969 TI - Patient-centred medicine through student-centred teaching: a student perspective on the key impacts of community- based learning in undergraduate medical education. AB - CONTEXT: The UK General Medical Council has proposed that increased use of community settings is essential to enhancement of medical education. However, such curriculum developments have been directed by educationalists and clinical faculty; there is to date little to show whether student perspectives accord with such expectations. AIM: To examine student views on whether community-based learning during a UK undergraduate medical education course results in new learning in the areas expected, and to elucidate any process factors which enhance attainment of learning objectives. METHOD: Nominal group technique, to develop consensus on important learning outcomes and process factors, and questionnaire survey, developed from the views of the nominal groups. RESULTS: 89 students participated (response rate 70% for the nominal groups, and 88% for questionnaire). Students perceived increased learning in many of the areas expected. In particular, students reported significant learning from: witnessing the impact of a longer term and more personal relationship with patients; the visible impact of social environment on health; the importance of dealing with people rather than diseases, and the use of the whole team for care. In addition, they emphasized that tutor, staff and patient enthusiasm for student presence and learning greatly enhanced the student learning experience. CONCLUSIONS: Community settings appear to achieve the expected attitudinal adaptation of students. The role of the committed tutor and team is seen as pivotal to learning. The conclusions support an increased emphasis in contemporary medical education and related research activity on the key impact of relationships in the learning environment. PMID- 11437970 TI - Students' evaluation of surgical seminars in a teaching hospital. AB - AIM: To evaluate the surgical seminars given for medical students at Auckland University and factors affecting them, so as to define areas for improvement. METHODS: A confidential questionnaire was completed by fourth-year medical students for each surgical seminar they attended during an academic year. The seminars were repeated four times during the year. The questionnaire consisted of five statements relating to whether the seminar was stimulating, problem- oriented, patient-centred, evidence-based and interactive. The students rated each statement on a 7-point scale. RESULTS: 556 questionnaires evaluating 28 clinical teachers were collected. A generalized linear model showed that the clinical teachers (F=4.16, P = 0.0001), the attachment (F=4.39, P=0.005) and the interaction between the seniority and institute (F=6.38, P=0.019) had a significant effect on the rating of the seminars. University consultants and hospital registrars had the highest overall least-squares mean, followed by hospital consultants, while university lecturers had the lowest overall least squares mean (5.56, 5.4, 5.18, and 4.36, respectively). Ratings for problem oriented approach improved over the studied period, while ratings of evidence based and patient-centred approaches remained lower. University consultants and hospital registrars had the best patient-centred approach compared with hospital consultants and university lecturers (least-squares means 5.52, 5.4, 5.01, 4.18, respectively). Hospital registrars showed the best interactive ability ratings (least-squares mean 5.51) while university lecturers had the least stimulating seminars (least-squares mean 4.46). CONCLUSIONS: Surgical seminars which were introduced to cover surgical topics had acceptable ratings in each of the teaching domains, although there is a need for improvement in the patient-centred and evidence-based approaches. PMID- 11437971 TI - Should medical students act as surrogate patients for each other? AB - Until recently, most clinical teachers and medical students have regarded using medical students as surrogate patients for peer teaching of physical examinations and clinical skills as practical and uncontroversial. Recent changes to medical curricula and changes in hospitalized patient populations have led to questions about the ethical acceptability of this practice. This paper explores the ethical issues inherent in the use of medical students as surrogate patients. It suggests that, ethically, there are parallels with two situations: when students conduct physical examinations on patients and when students participate as subjects in research. Drawing on accepted ethical practice in these two germane areas, the paper argues that there are both ethical strengths and weaknesses in the practice of using students as surrogate patients. Strategies to promote free and informed involvement of students as surrogate patients are suggested. PMID- 11437972 TI - What can students learn from studying medicine in literature? AB - OBJECTIVE: To identify what students can learn from studying medicine and literature as part of a final-year special study module in general practice. METHOD: The project reports and evaluation forms completed by all eight students who had chosen to study medicine and literature as part of their special study module in general practice at Nottingham University Medical School were analysed qualitatively. RESULTS: Students said that they gained a greater understanding of the patient's experience of illness, as a result of the texts they had read. They learned how illness can affect family or friends of the patient and about the psychological impact of physical illness. Most students thought their future care of patients would be influenced by what they had learned. Studying medicine in literature during a clinical attachment allowed students to draw comparisons between literature and their clinical experience. DISCUSSION: This study is based on a small number of students who chose to study medicine in literature. The results may not be generalizable to all medical students and not all students may be receptive to this method of learning. However, we recommend that students who are interested should be given the opportunity to study medicine in literature during their clinical years. This can enable them to reflect on their clinical experience and can provide a more profound understanding of the consequences of illness for the patient and their family. PMID- 11437973 TI - Use of a journal club and letter-writing exercise to teach critical appraisal to medical undergraduates. AB - INTRODUCTION: There is growing interest in methods of teaching critical appraisal skills at undergraduate and postgraduate levels. We describe an approach using a journal club and subsequent letter writing to teach critical appraisal and writing skills to medical undergraduates. METHODS: The exercise occurs during a 3 week public health medicine attachment in the third year of the undergraduate curriculum. Students work in small groups to appraise a recently published research paper, present their findings to their peers in a journal club, and draft a letter to the journal editor. Evaluation took place through: informal and formal feedback from students; number of letters written, submitted and published, and a comparison of marks obtained by students submitting a literature review assignment with and without critical appraisal teaching during the public health attachment. RESULTS: Feedback from students was overwhelmingly positive. In the first 3(1/2) years, 26 letters have been published or accepted for publication, and 58 letters published on the Internet. There were no significant differences in overall marks or marks for the critical appraisal component of the literature review assignments between the two student groups. DISCUSSION: We believe our approach is an innovative and enjoyable method for teaching critical appraisal and writing skills to medical students. Lack of difference in marks in the literature review between the student groups may reflect its insensitivity as an outcome measure, contamination by other critical appraisal teaching, or true ineffectiveness. PMID- 11437974 TI - Comparing health care delivery systems--initiating a student exchange project between Europe and the United States. AB - BACKGROUND: Cross-cultural contact among different health care systems can provide a framework for identifying the strengths and weaknesses of one's own healthcare system. However, such contact has rarely had much impact upon medical education curricula. Despite intense debate on reforming the healthcare delivery systems (HCDS) in Europe and the United States, there is very little formal representation of this interdisciplinary field in our educational programs. DESCRIPTION: To address this problem, a medical student exchange program was conducted in which students developed case studies that produced comparative analyses of HCDS in Germany, Sweden, Denmark and the United States. Each case is intended to highlight critical differences among the systems. EVALUATION: Students and their faculty preceptors completed pre- and post-exchange questionnaires to assess perceived knowledge of the HCDS and the adequacy of time devoted to it in their curricula. Both perceived that too little attention was devoted to this content in their programs. Following the exchange, students described clear increases in perceived knowledge. DISCUSSION: Our common interest in curriculum reform was key to implementing the exchange. The written cases generated by the students are being developed as course material in some of the schools and a conference is planned to disseminate the cases and the implementation strategies for their inclusion in medical curricula. PMID- 11437975 TI - Life in the fast lane: student life in the University of Hong Kong's new medical curriculum. PMID- 11437976 TI - Medical education in the new millennium--a Caribbean perspective. PMID- 11437977 TI - The juniors are revolting. PMID- 11437978 TI - Work-based learning (WBL). PMID- 11437979 TI - Neurogastroenterology and motility: new millenium, new horizons. PMID- 11437980 TI - Gastric emptying: the validity of the paracetamol absorption test adjusted for individual pharmacokinetics. AB - An algorithm for the paracetamol absorption test for gastric emptying, adjusting for individual pharmacokinetics, was recently developed. The aim of the present study was to validate the use of this algorithm. Furthermore, the algorithm was applied to elucidate whether a gastric tube interferes with the rate of gastric emptying. A caloric liquid meal with paracetamol was administered orally to nine healthy volunteers on two separate days. On one occasion, the subjects were intubated with a nasogastric tube and the meal was aspirated from the stomach 45 min after meal intake. The percentage of the meal retained in the stomach at the time of aspiration was determined by analyses of paracetamol in the aspirate and compared with calculations by the algorithm. On the other examination day, the same meal was ingested without tube and aspiration. The median percentage of the meal retained in the stomach at aspiration was 47% (range 33-70%) calculated by the algorithm and 48% (range 23-61%) based on the aspiration data. The correlation between the emptying parameters was r=0.97 (P < 0.001). The median of gastric emptying parameters was similar when the number of samples included in the calculation by the algorithm was reduced, but the range tended to increase. The gastric tube moderately inhibited gastric emptying during the period 20-40 min after meal intake (P < 0.05), but for the period from meal intake until start of aspiration, no inhibition was found. The present study demonstrates that the novel algorithm for the paracetamol absorption test provides valid estimates for gastric emptying. PMID- 11437981 TI - Neuronal control of the pyloric sphincter of the guinea-pig. AB - The pyloric sphincter (PS) controls gastric emptying and prevents the reflux of duodenal content into the stomach. Neuronal pathways and reflexes controlling the guinea-pig PS were physiologically investigated in isolated preparations. Simultaneous intracellular or extracellular and tension recordings from PS circular muscle with electrical and stretch stimulation were used. Electrical stimulation evoked an initial small contraction followed by a relaxation with a corresponding inhibitory junction potential (IJP) then a second large contraction with a corresponding excitatory junction potential (EJP). Hyoscine (1 micromol L 1) blocked the first contraction, and reduced the second contraction and EJP by 52.5% and 61%, respectively. These responses were further reduced by the NK2 antagonist, MEN10627 (1 micromol L-1), and the NK1 antagonist, SR140333 (1 micromol L-1). N-nitro-L-arginine (100 micro;mol L-1) and apamin (0.5 micromol L 1) blocked the relaxation and the IJP. Duodenal electrical stimulation evoked an EJP, whereas antral stimulation evoked an IJP followed by a small EJP. All were blocked by hexamethonium (100 micromol L-1). Duodenal stretch evoked tetrodotoxin sensitive reflex contractions and membrane depolarization with action potentials in the PS. Thus, PS enteric motor neurones receive inputs from the duodenum and the stomach. There are stretch-sensitive ascending excitatory reflex pathways from the duodenum to the PS. PMID- 11437982 TI - Oesophageal pressure-cross-sectional area distributions and secondary peristalsis in relation to subclassification of systemic sclerosis. AB - The aim of the present study was to correlate the severity of oesophageal motor dysfunction with the severity of cutaneous disease in systemic sclerosis (SS). Patients were divided into three groups based on the degree of skin involvement: type I, acrosclerosis distal to the wrist; type II, scleroderma extending above the wrist in proximal direction; type III, diffuse cutaneous systemic sclerosis. Impedance planimetry employing distensions with pressures up to 5 kPa with the concomitant measurement of oesophageal cross-sectional area (CSA) was used in combination with standard oesophageal manometry. Measurements were made at 7 and 15 cm above the lower oesophageal sphincter (LOS). Thirty patients (16 type I, six type II and eight type III patients) and 23 normal controls were included. LOS pressure was lower in SS patients than in normal patients, with the lowest values in type III. The CSAs were higher in SS patients than in controls at both sites (P < 0.001). The CSAs at the distal site were highest in type III, as compared to type I and II (P < 0.03). The CSA at the highest induced pressure (5.0 kPa) was 613 +/- 45, 719 +/- 79, and 808 +/- 115 mm2 in types I, II and III, respectively. No differences in CSA were found at the proximal site between the three types of SS. The distensibility did not differ between SS and normal patients at the distal site. The distensibility was lowest in SS patients (P < 0.001) at the proximal distension site. The distensibility did not vary with the type of SS at either site. Significant differences in contraction frequency of the secondary peristalsis as function of wall tension were demonstrated between the SS patients and controls at the distal site (P < 0.05). No differences were found at the proximal site. The contraction frequency and amplitude at the distal and proximal sites did not differ among the three types. In conclusion for most parameters studied, SS patients differed from normal patients. Among SS types, the most pronounced changes were found in type III. PMID- 11437983 TI - Contribution of NK3 tachykinin receptors to propulsion in the rabbit isolated distal colon. AB - The role of NK3 receptors in rabbit colonic propulsion has been investigated in vitro with the selective agonist, senktide, and two selective antagonists, SR142801 and SB222200. Peristalsis was elicited by distending a rubber balloon with 0.3 and 1.0 mL of water leading to a velocity of 2.2 and 2.8 mm s-1, respectively. At concentrations of 1 nM, senktide inhibited propulsion evoked by both distensions (range 25-40%), whereas at 6 and 60 nmol L-1 facilitated 'submaximal' propulsion by 30%. In the presence of Nomega-nitro-L-arginine (L NNA, 200 micromol L-1), which per se caused a slight prokinetic effect, 1 nmol L 1 senktide markedly accelerated propulsion (range 35-50%). Hexamethonium (200 micromol L-1) had minor effects on propulsion. In its presence, 60 nmol L-1 senktide significantly inhibited propulsion induced by both stimuli (range 20 50%). SR142801 (0.3, 3 nmol L-1) and SB222200 (30, 300 nmol L-1) facilitated 'submaximal' propulsion (range 20-40%). Conversely, higher antagonist concentrations (SR142801: 30, 300 nM; SB222200: 1, 10 micromol L-1) inhibited propulsion to both distensions by 20%. A combination of SR142801 (300 nmol L-1) plus hexamethonium (200 micromol L-1) induced an approximately four-fold greater inhibition of propulsion than that induced by SR142801 alone. In conclusion, in the rabbit-isolated distal colon, a subset of NK3 receptors located on descending pathways mediates an inhibitory effect on propulsion by activating a NO-dependent mechanism. Another subset of NK3 receptors, located on ascending pathways mediates a facilitative effect involving a synergistic interaction with cholinergic nicotinic receptors. PMID- 11437984 TI - Endogenous nitric oxide release modulates the direction and frequency of colonic migrating motor complexes in the isolated mouse colon. AB - Spontaneous colonic migrating motor complexes (CMMCs) were recorded from circular muscle at three sites along the isolated mouse colon. The interval between CMMCs was decreased from approximately 3 min in control solution, by approximately 55% in a nitric oxide synthase (NOS) inhibitor, N-nitro-L-arginine (L-NNA; 100 micromol L-1). This was associated with a shift in migration direction of CMMCs, such that CMMCs migrated in an oral direction. Application of the endogenous substrate for NOS, L-arginine, at a low concentration used to mimic plasma concentration (134 micromol L-1), or a high concentration (5 mmol L-1) suppressed CMMCs (for at least 15 min) which were replaced by high frequency (10-15 min-1), short duration (half width approximately 1.5 s) contractions of variable amplitudes (largest in the proximal region) until CMMCs resumed. CMMCs remained in the presence of D-arginine (134 micromol L-1 and 5 mmol L-1). Apamin (250 nmol L-1) did not alter the interval between CMMCs, however, additional nonmigrating contractions were observed between the CMMCs in the distal region. In addition to its effects on smooth muscle tone, NO, but not apamin-sensitive channels, plays an important role in suppressing the frequency of migrating contractions in the isolated mouse colon. Consideration should be given to the inclusion of L arginine, in in vitro experiments where there may be spontaneous activity in NOS containing neurones. PMID- 11437985 TI - Involvement of spinal calcitonin gene-related peptide in the development of acute visceral hyperalgesia in the rat. AB - This study aimed to characterize the role of the neuropeptide calcitonin gene related peptide (CGRP) in the development of mechanically induced visceral hyperalgesia. Tonic colorectal distension (CRD) was performed in fasted, conscious male Sprague-Dawley rats. The visceromotor reflex associated with noxious CRD was determined as the number of contractions during each of two consecutive tonic distensions (10 min at 60 mmHg), which were separated by a series of phasic distensions (repeated 15-s distensions to 80 mmHg at 30-s intervals). The effect of the CGRP receptor antagonist h-CGRP8-37 given intrathecally (i.t.) (0.03-3 nmol rat-1) or intravenously (i.v.) (20 microg kg-1 bodyweight [bw]) on the visceromotor response was evaluated. The dose for i.v. administration was chosen based on previous results from similar studies. In addition, the effect of a CGRP monoclonal antibody (6 mg kg-1 bw) given intravenously was evaluated. Compared to the baseline response, a significant increase in the number of abdominal contractions was observed during the second tonic distension. The i.t. application of h-CGRP8-37 dose-dependently reduced the numbers of abdominal contractions both during the first and the second tonic distension period, with a maximum effect observed at a peptide concentration of 3 nmol. Intravenous administration of h-CGRP8-37 or of the CGRP antiserum produced a small reduction of the visceromotor response induced by the second tonic distension and had no effect on colonic compliance. The development of mechanically induced colorectal hyperalgesia by repeated tonic distension involves the spinal release of CGRP, while peripheral release of CGRP plays only a minor role. PMID- 11437986 TI - Phase III of the migrating motor complex: associated with endogenous xenin plasma peaks and induced by exogenous xenin. AB - Xenin, a recently discovered peptide produced by specific endocrine cells of the duodenal mucosa, has shown exocrine, endocrine and motility effects in the gastroenteropancreatic system in animal experiments. The aim of the present investigation was to study the role of xenin in the regulation of duodenojejunal motility of humans. Twenty-nine healthy volunteers from the hospital staff gave informed consent to participate in this investigation. In 20 volunteers, we determined plasma concentrations of immunoreactive xenin at 15 min intervals over a mean time period of 8 h fasting and recorded the interdigestive motor activity of the duodenojejunum. In a double-blind randomized crossover study on other nine subjects, synthetic xenin in a dose of 4 pmol kg-1 min-1 or placebo was infused for 10 min intravenously in the interdigestive period and postprandially after a liquid meal. Duodenojejunal motility was recorded simultaneously. Predefined interdigestive xenin plasma peaks were found to be significantly associated with the phases III of the migrating motor complex. In the interdigestive period, xenin induced a premature phase III activity in each volunteer; this was followed by a second phase III in five out of nine subjects. In the postprandial state, xenin significantly increased contraction frequency and the percentage of aborally propagated contractions. These findings suggest a role of the peptide hormone xenin in modulating interdigestive and postprandial duodenojejunal motility in humans. PMID- 11437987 TI - Galanin suppresses calcium conductance and activates inwardly rectifying potassium channels in myenteric neurones from guinea-pig small intestine. AB - Whole-cell patch-clamp recording methods were used to investigate the ionic mechanisms underlying the hyperpolarizing action of galanin in enteric neurones. Galanin suppressed calcium current (ICa) and activated inwardly rectifying potassium current (IK,ir) in AH-type myenteric neurones of guinea-pig small intestine. Both suppression of ICa and activation of IK,ir were concentration dependent, with an EC50 of 1.4 nmol L-1 and 55 nmol L-1, respectively. Pretreatment with pertussis toxin eliminated both actions of galanin, suggesting that both galanin-induced inhibition of ICa and galanin-induced activation of IK,ir involved activation of Gi/Go proteins. Both suppression of ICa and activation of IK,ir by galanin were mimicked by the N-terminal fragment of galanin, galanin-(1-16) suggesting that the first 16 amino acids of the peptide were sufficient for both actions. The galanin receptor antagonist galantide suppressed the galanin-induced activation of IK,ir with an EC50 of 16 nmol L-1. However, galantide alone suppressed ICa. The results suggest two mechanisms of action for galanin: one is opening of inwardly rectifying potassium channels and the second is blockade of voltage-activated calcium channels. PMID- 11437988 TI - Neurotransmitter coding of enteric neurones in the submucous plexus is changed in non-inflamed rectum of patients with Crohn's disease. AB - Knowledge of the neurochemical coding of submucosal neurones in the human gut is important to assess neuronal changes under pathological conditions. We therefore investigated transmitter colocalization patterns in rectal submucosal neurones in normal tissue (n=11) and in noninflamed tissue of Crohn's disease (CD) patients (n=17). Neurone-specific enolase (NSE), choline acetyltransferase (ChAT), vasoactive intestinal polypeptide (VIP), substance P (SP), nitric oxide synthase (NOS) and calcitonin gene-related peptide (CGRP) were detected immunohistochemically in whole-mount preparations from rectal biopsies. The neuronal marker NSE revealed no differences in the number of cells per ganglion (controls 5.0; CD 5.1). Four cell populations with distinct neurochemical codes were identified. The sizes of the populations ChAT/VIP (58% vs. 55%), ChAT/SP (8% vs. 8%), and ChAT/- (22% vs. 22%) were similar in control and CD. The population VIP/- was significantly increased in CD (12% vs. 2% in controls). Unlike in controls, all NOS neurones colocalized ChAT in CD. Thickened CGRP-fibres occurred in CD. We identified neurochemically distinct populations in the human submucous plexus. The increase in the VIP/- population, extensive colocalization of ChAT and NOS and hypertrophied CGRP fibres indicated adaptive changes in the enteric nervous system in noninflamed rectum of CD patients. PMID- 11437989 TI - Damage control--a possible non-proteolytic role for ubiquitin in limiting neurodegeneration. AB - Ubiquitin can be detected in the neuronal and glial inclusions that are the diagnostic hallmarks of a number of human neurodegenerative diseases. It has been assumed that the presence of ubiquitin signifies the failed attempt of the cell to remove abnormal protein structures, which have been allowed to aggregate. The burden of abnormal protein arising from genetic mutations or cumulative oxidative damage might in the course of time overwhelm the ubiquitin-proteasome pathway (whose responsibility it is to eliminate misfolded or damaged proteins). However, ubiquitin may still serve a protective purpose distinct from its role in proteolysis. The physical properties of ubiquitin are such that a surface coating of ubiquitin should preclude further growth of the aggregate, prevent non productive interactions, and conceal the contents from detection mechanisms that might ultimately kill the cell. This 'nonstick coating' hypothesis makes predictions about the nature of the conjugated ubiquitin and the consequences of removing it. PMID- 11437990 TI - Ubiquitin C-terminal hydrolase-L1 (PGP9.5) expression in human neural cell lines following induction of neuronal differentiation and exposure to cytokines, neurotrophic factors or heat stress. AB - Dysfunction of the ubiquitin-dependent proteolytic pathway contributes to progressive accumulation of ubiquitinated protein inclusions in neurodegenerative disorders, such as Parkinson's disease (PD). Ubiquitin C-terminal hydrolase-L1 (UCH-L1), alternatively designated protein gene product 9.5 (PGP9.5), is a neural deubiquitinating enzyme which is identified as a principal constituent of Lewy bodies. To clarify the regulatory mechanism of UCH-L1 expression in human neural cells, we studied the constitutive, cytokine/neurotrophic factor-regulated, and heat stress-induced expression of UCH-L1 in cultured human neural cell lines by Western blot analysis. The constitutive expression of UCH-L1 was identified in SK N-SH neuroblastoma cells, IMR-32 neuroblastoma cells, U-373MG astrocytoma cells, and NTera2 teratocarcinoma-derived differentiated neurones (NTera2-N). The levels of UCH-L1 expression were unaltered in these cell lines following treatment with TNF-alpha, IL-1beta, BDNF, GDNF, dibutyryl cyclic AMP, or phorbol 12-myristate 13 acetate, and remained unchanged by exposure to heat stress. In contrast, its levels were elevated substantially in NTera2 teratocarcinoma cells following retinoic acid-induced neuronal differentiation, accompanied with an increased expression of alpha-synuclein and synaptophysin. These results indicate that UCH L1 is expressed constitutively in human neual cell lines, where it is upregulated following induction of neuronal differentiation, but unaffected by exposure to heat stress, cytokines, or growth/differentiation factors which are supposed to be invloved in the nigral neuronal death and survival in PD. PMID- 11437991 TI - Mutational analysis of TSC1 and TSC2 genes in gangliogliomas. AB - Gangliogliomas constitute the most frequent tumour entity in patients with temporal lobe epilepsy. The characteristic histopathological admixture of glial and neuronal elements, the focal nature and their differentiated phenotype and benign biological behaviour suggest an origin from a developmentally compromised or dysplastic precursor lesion. The present study analysed TSC1 and TSC2 genes as potential candidates involved in the pathogenesis of this intriguing neoplasm. Recent data suggest that both genes play a role in cortical differentiation and growth control. DNA sequence analysis of TSC1 and TSC2 was studied in 20 patients with gangliogliomas. Fifteen of these tumours (75%) carried polymorphisms in the TSC2 gene. The frequency of these polymorphisms was significantly increased in intron 4 (12.5%) and exon 41 (15%) compared to control individuals (8.1 and 6.5%, respectively, n = 100). A somatic mutation in intron 32 of the TSC2 gene was encountered in one patient. In the TSC1 gene, seven polymorphisms occurred as a combination of base exchanges in exon 14 and intron 13. No mutations were observed in this gene. Laser microdissection and harvesting of individual neuronal and glial elements identified the intron 32 mutation within the glial portion but not in dysplastic neurones of the tumour. The data demonstrate numerous polymorphisms as well as a novel TSC2 mutation in gangliogliomas from patients with chronic epilepsies. The selective detection of the TSC2 mutation within the glial component of a ganglioglioma suggests that the glioma portion has undergone clonal evolution in this case. PMID- 11437992 TI - Alterations in neurofilaments associated with reactive brain changes and axonal sprouting following acute physical injury to the rat neocortex. AB - In order to study the changes in axons related to acute localized physical trauma, a 25 gauge needle was inserted into the somatosensory cortex of anaesthetized adult rats. Animals were examined over 11 time points, from 30 min to 14 days postinjury. Initially, the central needle tract was surrounded by 'reactive' abnormal axons characterized by their bulb- or ring-like immunoreactivity for neurofila ments. Quantification demonstrated that these structures reached a peak density at 24 h postinjury, followed by a gradual decrease over 2 weeks. By 5 days postinjury, long axons showing high levels of neurofilament labelling were localized to the lesion area, either aligned parallel to the tract edges or extending into the bridge of tissue forming between the tract edges. Double-labelling demonstrated a close association between sprouting axons and ferritin-labelled microglia. Immunolabelling for GAP43 also demonstrated the presence of sprouting axons within this tissue bridge. Ultrastuctural examination showed that sprouting axons contained a high density of neurofilaments, with a leading edge lacking these filaments. Injury to the adult neocortex is associated with reactive and sprouting changes within axons, coordinated with the proliferation of microglia and wound healing. These data also support a role for neurofilaments in axonal sprouting following brain injury. PMID- 11437993 TI - CXCR3 expression in human central nervous system diseases. AB - The CXCR3 chemokine receptor, expressed on activated T lymphocytes, is seen within the central nervous system (CNS) in inflammatory conditions where a T-cell response is prominent. However, the distribution of CXCR3 in parenchymal CNS cells is unknown. Using a monoclonal antibody against CXCR3 and post-mortem tissue of patients with and without CNS pathology, we have determined its expression pattern. CXCR3 was found in subpopulations of cells morphologically consistent with astrocytes, particularly reactive astrocytes, and in cerebellar Purkinje cells. It was also detected in arterial endothelial and smooth muscle cells, particularly in areas associated with atherosclerotic plaques. CXCR3 positive astrocytes were particularly prominent in the CNS of HIV-positive patients, in patients with Multiple Sclerosis (MS), in ischaemic infarcts and in astrocytic neoplasms. Immunofluorescence studies of mixed adult primary glial cultures and fetal glial cultures also showed expression of CXCR3 in astrocytes. CXCR3 mRNA was detected in Purkinje cells by in situ hybridization with a CXCR3 specific probe. Thus, the predominant expression of CXCR3 in reactive astrocytes may indicate that it plays a role in the development of reactive gliosis in a variety of infectious, inflammatory, vascular and neoplastic processes in the CNS. The relationship between CXCR3 expression in astrocytes to its expression in Purkinje cells, endothelial cells and smooth muscle cells is yet to be determined. PMID- 11437994 TI - An unusual complication of ADEM. PMID- 11437995 TI - A web page to teach neurology and neuropathology to medical students. PMID- 11437996 TI - Erythema toxicum neonatorum: an immunohistochemical analysis. AB - Erythema toxicum neonatorum is a benign rash of unknown etiology, present to various degrees in most term newborns and characterized by an accumulation of eosinophils in dermal lesions. The recruitment of leukocytes to tissues implicates the involvement of adhesion molecules, cytokines, and chemokines. We therefore performed immunohistochemistry on punch biopsy specimens from cutaneous lesions of ten 1-day-old infants with erythema toxicum using specific monoclonal antibodies directed against a variety of adhesion molecules, cytokines, chemokines, and cell type-specific membrane markers. Biopsy specimens of noninflamed skin from four matched newborns and four adults served as controls. The immunohistologic features of erythema toxicum in all 10 infants included a strong staining of the adhesion molecule E-selectin in the vessel wall and the presence of numerous inflammatory cells that were identified as dendritic cells (CD1a, CD83, HLA-DR, CD40, and ICAM-1 positive), eosinophils (EG2 positive), neutrophils (CD15 positive), macrophages (CD14, CD68, and Mac387 positive), and E selectin-expressing cells. Furthermore, the lesions showed a high incidence of the proinflammatory cytokines interleukin (IL)-1alpha and IL-1beta and of the chemokines IL-8 and eotaxin. This immunologic activity was reduced or absent in noninflamed skin from newborn controls and adults. We conclude that there is an accumulation and activation of immune cells in the lesions of erythema toxicum, also present in noninflamed skin of 1-day-old infants, but to a lower level. The physiologic significance of the rash remains to be elucidated. PMID- 11437997 TI - Childhood psoriasis: a clinical review of 1262 cases. AB - Our aim was to describe the types of psoriasis seen in a large series of patients presenting to a tertiary referral pediatric dermatology department using a classification system combining conventional terminology and additional categories based on the site and characteristics of the rash. A total of 1262 patients seen consecutively in the dermatology department of the Royal Alexandra Hospital for Children, Sydney, Australia, between 1981 and 1995 are described and classified according to the pattern of psoriasis at the time of presentation. Additional information recorded included family history, facial involvement, and history of a psoriatic type of diaper rash in infancy. The ages of the children ranged from 1 month to 15 years. There was an equal gender distribution and a high rate of positive family history at 71%. Twenty-six percent of children had a history of a psoriatic diaper rash and facial involvement occurred in 38% of children. Plaque psoriasis was the most common type overall, affecting 430 patients (34%). Three hundred forty-five children were less than 2 years of age, and this is the largest series of children with psoriasis in this age group presented to date. An entity defined by us as psoriatic diaper rash with dissemination was the most common type of psoriasis in the less than 2-year age group, affecting 155 (45%) patients. This large series offers information on the manifestations of psoriasis in childhood, but is particularly useful in examining the previously less well-described infant age group. The classification used is proposed as a practical way to describe psoriasis in children, particularly with respect to future descriptive studies. PMID- 11437998 TI - Cutaneous manifestations of chronic renal failure in children of color. AB - One hundred percent of adult patients with chronic renal failure (CRF) develop cutaneous findings as a result of uremia or due to therapeutic interventions. To date, pediatric incidence studies have been limited to Caucasian children. However, recent reports have indicated that more African American patients progress to end-stage renal disease (ESRD). This is the first study to assess the prevalence of renal failure-related skin disease in children of color, including African American and Hispanic patients. Thirty children were evaluated by history and physical examination, with assignment to one of three treatment categories: transplanted (n = 10), dialyzed (n = 16), or medically managed (n = 4). Skin findings were divided into uremic, drug-related, or infectious disease types. The incidence of skin disease was 100%. Xerosis was the single most common finding, often accompanied by pruritus. Cushinoid features were common despite the addition of steroid-sparing agents. Cyclosporin A-treated African American children had a high incidence of gingival hypertrophy (72%) and an even higher incidence of hypertrichosis (100%). Acral warts and nevi were common findings, the latter correlating with the length of immunosuppression. There is a high incidence of cosmetically disfiguring side effects (Cushinoid facies, hypertrichosis, and gingival hypertrophy) in children within all treatment categories, primarily related to drug treatment. Further study is required to determine the long-term sequelae, including psychological disturbances, of cutaneous disease in children of color with CRF. PMID- 11437999 TI - Cutaneous complications of BCG vaccination in infants with immune disorders: two cases and a review of the literature. AB - Two infants, one with a T-cell-signaling defect resulting in a primary immunodeficiency syndrome and the other with severe combined immunodeficiency (SCID), are described. Both infants developed cutaneous infections secondary to their bacillus Calmette-Guerin (BCG) vaccinations. Both patients were from countries where BCG is routinely administered in infancy. The infant with the T cell-signaling defect developed a disseminated infection involving the skin, while the infant with SCID developed a localized cutaneous infection at the site of his BCG immunization. These two cases resemble other reported cases of cutaneous BCG infection following routine vaccination in immunocompromised patients. Mycobacterium bovis infection should be considered in patients with cutaneous eruptions who have received BCG vaccination, especially those who are immunocompromised. PMID- 11438000 TI - Rothmund-Thomson syndrome with myelodysplasia. AB - Rothmund-Thomson syndrome (RTS) is a rare autosomal recessive disorder that is caused by a DNA repair defect. It is characterized mainly by skin, eye, and skeletal abnormalities. Cutaneous changes appear at between 3 and 6 months of age and include poikiloderma, photosensitivity, scaling, hyperkeratosis, and disturbance of hair growth. Other abnormalities include cataracts, congenital bone defects, soft tissue contractures, and osteogenesis imperfecta. Various malignancies have been reported in association with RTS, including osteosarcoma, fibrosarcoma, and nonmelanoma skin cancers. The myelodysplastic syndromes are a group of hematologic disorders defined by morphologic abnormalities of the three cell lines. The pathogenesis of myelodysplasia is a multistep process that begins with a somatic mutation in the pluripotential stem cell, which is irreversibly altered and acquires a survival advantage. Myelodysplasia in the young and RTS are both rare conditions. We report a patient with RTS and myelodysplasia. This is the second reported case of an association between these two conditions, which are both likely to be due to a common etiologic cause of nonrepair of stem cell DNA damage. Clinicians should be aware of the potential of this complication arising in patients with RTS. PMID- 11438001 TI - Generalized acanthosis nigricans in early childhood. AB - Acanthosis nigricans (AN) is a disorder often accompanied by internal malignancy in adult patients, but in children it is usually benign. Generalized forms of AN, especially in childhood, are extremely rare. We report a 5-year-old boy with a 3 year history of AN who had generalized roughness and hyperpigmentation, numerous skin wrinkles, disseminated skin tags, tripe palms, and intensive pruritus. No clinical evidence of internal disorders, notably of an endocrinologic or neoplastic nature, have been detected thus far. PMID- 11438002 TI - Herlitz junctional epidermolysis bullosa presenting at birth with anonychia: a case report and review of H-JEB. AB - A 15-day-old Yemeni boy presented with anonychia and granulomatous nail beds and white patches in the mouth. Biopsy specimens from the nail beds were nondiagnostic. Shortly thereafter the child developed multiple tense bullae, a hoarse voice, and poor appetite. Hematoxylin and eosin staining along with monoclonal antibody studies of a skin biopsy specimen revealed subepidermal bullae through the lamina lucida and a marked decrease in laminin 5. A diagnosis of junctional epidermolysis bullosa Herlitz variant was made. His course was complicated by multiple nonhealing wounds, oral pharyngeal involvement, sepsis, anemia, and poor nutrition, leading to his eventual death. This report emphasizes the unusual presentation of Herlitz junctional epidermolysis bullosa with anonychia as the initial finding and a relatively prolonged period before cutaneous blister formation, resulting in delay of diagnosis. PMID- 11438003 TI - Congenital plaquelike glomus tumor in a child. AB - An 8-year-old girl was seen in our outpatient clinic with a history of an angiomatous plaque on the chest since birth. She had complained of pain at the site of the lesion for 1 month. Histopathologic examination and immunohistochemical analysis led to the diagnosis of glomangioma, a rare congenital plaquelike glomus tumor. We review the literature and discuss the clinical presentation and evolution of this rare entity. PMID- 11438004 TI - Subepidermal calcified nodule: report of two cases and review of the literature. AB - Subepidermal calcified nodule (SCN) is a form of calcinosis cutis that most commonly occurs in children. The typical clinical presentation is of a solitary verrucous nodule on the face, but occasionally multiple lesions may be seen. We describe SCN on the eyelids of two female patients; one of whom had four lesions. The histopathology was typical of calcinosis cutis. Both patients were in good health. PMID- 11438005 TI - Familial atrophia maculosa varioliformis cutis: an ultrastructural study. AB - Atrophia maculosa varioliformis cutis is a rare and distinctive form of idiopathic facial macular noninflammatory atrophy that may rarely be observed in members of the same family. We describe two brothers, ages 14 and 16 years, with spontaneously appearing, asymptomatic, varioliform and linear atrophic lesions. Their past medical history was positive for varicella occurring in childhood without residual facial scarring. Routine laboratory investigations and screening for circulating autoantibodies were negative. Both patients were concordant for HLA A2 and DQ4.1. Routine and ultrastructural histologic examination of a punch biopsy specimen showed the presence of scarce, small, fragmented elastic fibers and compact collagen bundles associated with hypertrophic fibroblasts in the dermis. Our patients remained clinically stable, untreated, over a 2-year follow up period. No long-term follow-up data have previously been reported. PMID- 11438006 TI - Congenital granular cell tumors localized to the arm. AB - We report an instance of congenital granular cell tumors localized to the arm of a female infant. While granular cell tumors are well described during infancy as congenital epulis of the oral cavity, this case is unusual in both its location and histologic characteristics. The lesions, located around the antecubital fossa, were comprised of CD34-positive, S-100-negative granular cells. In addition, there were numerous eccrine glands in the upper dermis. The salient features of the case are discussed and reviewed in the context of the literature pertaining to this unusual entity. PMID- 11438007 TI - Subepidermal calcified nodule. AB - Subepidermal calcified nodule is an uncommon, but specific subtype of idiopathic calcinosis. It presents as an asymptomatic, solitary, yellow-white or erythematous filiform tumor. The most common location is on the head and neck region of male children. Serum calcium and phosphorus levels are normal. Histopathologic examination reveals a focal papillary dermal collection of dark blue-staining, large, amorphous deposits, and/or small calcified globules surrounded by a lymphohistiocytic infiltrate. The causes and histiogenesis of these deposits have not been fully established, but the most favored theory is that calcium is deposited on a preexisting lesion. The treatment of choice is surgical removal with histopathologic examination. We describe a healthy 11-year old Caucasian boy with an asymptomatic, warty growth below his left eye. Excision with histopathologic examination confirmed the diagnosis of subepidermal calcified nodule. There has been no sign of recurrence to date. PMID- 11438008 TI - Combined treatment with calcipotriol ointment and low-dose ultraviolet A1 phototherapy in childhood morphea. AB - Various therapies for morphea have been used with limited success, including ones with potentially hazardous side effects. When morphea occurs in childhood it may lead to progressive and long-lasting induration of the skin and subcutaneous tissue, growth retardation, and muscle atrophy. We report an open prospective study in which the efficacy of a combined treatment with calcipotriol ointment and low-dose ultraviolet A1 (UVA1) phototherapy in childhood morphea was investigated. Nineteen children (mean age 8.5 years, range 3-13 years) with morphea were exposed to UVA1 (340-400 nm) phototherapy at a dose of 20 J/cm(2) four times a week for 10 weeks. Forty phototherapy sessions resulted in a cumulative dose of 800 J/cm(2) UVA1. In addition, calcipotriol ointment (0.005%) was applied twice a day. After 10 weeks, palpation and inspection showed a remarkable softening and repigmentation of formerly affected skin resulting in a highly significant (p < 0.001) decrease of the mean clinical score from 7.3 +/- 0.9 at the beginning to 2.4 +/- 0.9 (relative reduction 67.1%) at the end of combined therapy. Our results indicate that a combined therapy with calcipotriol ointment and low-dose UVA1 phototherapy is highly effective in childhood morphea. Further controlled studies are necessary to investigate whether this combined therapy is superior to UVA1 phototherapy alone. PMID- 11438009 TI - Low dose cyclosporin A treatment in generalized pustular psoriasis. AB - Generalized pustular psoriasis is a rare form of psoriasis, seldom seen in children. Three patients with generalized pustular psoriasis are presented, two of whom were a sister and brother and whose grandfather also had pustular psoriasis. Lesions consisted of pustular, erythematous, scaly, follicular papules located on the trunk, scalp, and extremities. The pustules in some areas coalesced to form lakes. Histologic examination of several biopsy specimens revealed the changes of pustular psoriasis, which were parakeratosis, elongation of the rete ridges, and deep spongioform pustules and Munro abscesses. All patients were treated with cyclosporin A for periods of 2-12 months. The doses ranged from 1 to 2 mg/kg/day. Clearance of psoriatic lesions occurred after 2-4 weeks of therapy. PMID- 11438010 TI - Raynaud phenomenon after treatment of verruca vulgaris of the sole with intralesional injection of bleomycin. AB - Intralesional injection of bleomycin (BLM) for the treatment of warts resistant to all conventional therapies is of certain interest because of the drug's low toxicity and its efficacy. However, delayed side effects may appear, particularly Raynaud phenomenon. Accordingly, some precautions must be taken in patients with vascular or collagen diseases and, as a precaution, pregnant or lactating women and infants should not be treated with this drug. PMID- 11438011 TI - What syndrome is this? CHIME syndrome. PMID- 11438012 TI - Vulvar mass in a toddler. PMID- 11438013 TI - Diagnosing eruptive vellus hair cysts. PMID- 11438014 TI - Striae distensae (rubrae) in monozygotic twins. PMID- 11438016 TI - Proceedings of the 1st workshop on the epidemiological survey of chronic obstructive lung diseases and alpha-1-antitrypsin deficiency in the Asian-Pacific region. 2 December 2000, Tokyo, Japan. PMID- 11438017 TI - Chronic obstructive pulmonary disease in Hong Kong. PMID- 11438018 TI - Epidemiological survey of chronic obstructive pulmonary disease and alpha-1 antitrypsin deficiency in Korea. PMID- 11438019 TI - Epidemiological survey of chronic obstructive pulmonary disease and alpha-1 deficiency in China. PMID- 11438020 TI - The epidemiology and clinical management of chronic obstructive pulmonary disease in Singapore. PMID- 11438021 TI - Epidemiological survey of chronic obstructive pulmonary disease and alpha-1 antitrypsin deficiency in Australia. PMID- 11438022 TI - Epidemiological survey of chronic obstructive pulmonary disease in Japan. PMID- 11438023 TI - State of alpha1-antitrypsin deficiency in Japan. PMID- 11438024 TI - The role of alpha-1-antitrypsin in generating chronic obstructive pulmonary disorder. PMID- 11438026 TI - Including polymerase chain reaction in screening for hepatitis C virus RNA in blood donations is not cost-effective. AB - BACKGROUND AND OBJECTIVES: To assess the cost and effectiveness of adding a system of polymerase chain reaction (PCR) testing to the current enzyme-linked immunosorbent assay (ELISA) screening for hepatitis C virus (HCV) in blood donations. MATERIALS AND METHODS: We performed a cost-effectiveness analysis comparing three HCV screening strategies in French blood donors: detection of antibodies against HCV by an ELISA on blood donations (the current policy) vs. ELISA and PCR either on each blood donation or on blood pools. We combined national surveillance data on HCV antibody-positive blood donors with a literature review in a Markov model for natural history of HCV disease and its treatment. RESULTS: PCR testing performed in parallel with ELISA will potentially add less than 1 year of life for all French recipients compared to ELISA screening alone. The incremental costs per life year saved were, respectively, epsilon84.6 million with ELISA and PCR testing of blood pools, and epsilon891.1 million with testing of individual blood donations. CONCLUSION: PCR testing performed in parallel with antibody screening, either on pools or on single donations is not currently a cost-effective option for the detection of HCV RNA virus in blood donors. PMID- 11438027 TI - National Institute for Biological Standards and Control/UK Blood Transfusion Service working standards for HBsAg, anti-HCV and anti-HIV-1 ('go/no-go' controls). AB - BACKGROUND AND OBJECTIVES: The positive controls supplied with the commercial kits used in the testing of blood donations for markers of viral infection are usually of high reactivity and do not give users a reliable indication of optimal performance of the kit on a day-to-day basis. MATERIALS AND METHODS: The National Institute for Biological Standards and Control has developed working standards for hepatitis B surface antigen (HBsAg), anti-hepatitis C virus (anti-HCV) and anti-human immunodeficiency virus type 1 (anti-HIV-1) for the UK Blood Transfusion Services for daily use in every test. RESULTS: Data from the use of these reagents are collated and quarterly reports are issued. CONCLUSIONS: The centralized collation, analysis and reporting of the results from users of working standards facilitate the continual assessment of the responses obtained by individual laboratories, comparisons of the interlaboratory performance and the performance of different manufacturers' kits at a national level. This information complements the data on infection surveillance and initial/repeat reactivities using the same kits. PMID- 11438028 TI - Detection of enterovirus viraemia in blood donors. AB - BACKGROUND AND OBJECTIVES: The infrastructure established for screening blood donations for hepatitis C virus has enabled large-scale population testing for other viruses which are potentially transmissible by transfusion of blood components and plasma-derived blood products. We have measured the frequency of viraemia of enteroviruses and parechoviruses in 83 600 Scottish blood donors to allow an initial assessment of their risk to blood safety. MATERIALS AND METHODS: Plasma samples collected from blood donors over 7 calendar months were tested anonymously in mini-pools of 95 donations, by polymerase chain reaction (PCR) for human enterovirus and parechovirus sequences. RESULTS: A total of 19 mini-pools, from the 880 that were tested, were PCR-positive for enterovirus RNA, predicting a donor prevalence of 0.023%. Enterovirus sequences were not detected in factor VIII or IX clotting factor concentrates. None of the 230 mini-pools or concentrates contained detectable parechovirus RNA. CONCLUSIONS: The prevalence of enterovirus viraemia detected in this study predicts that at least 1000 enterovirus-contaminated blood components are transfused per year in the UK. The frequency of transmission and clinical outcome after exposure to enterovirus contaminated blood components in recipients is unknown. PMID- 11438029 TI - Preparation of lyophilized and liquid intravenous immunoglobulin G: development and scale-up. AB - BACKGROUND AND OBJECTIVES: Intravenous immunoglobulin G (IVIG) preparations are used in several disorders: primary and secondary immunodeficiencies, autoimmune and systemic inflammatory diseases, and in infectious diseases as effective therapy. In this work, we report a method of processing IVIG (lyophilized and liquid) from Cohn fraction II (FII) paste. MATERIALS AND METHODS: The process included the following steps: virus inactivation by heat in a solution of FII, ultrafiltration, treatment at pH 4 with traces of pepsin, diafiltration and final formulation. Optimizations for each step on a small scale are described. The method has been effectively scaled-up. RESULTS: The final product contained at least 99% monomers plus dimers of IgG, a low anti-complementary activity (AcA) < 1 CH50/mg, and the remaining quality parameters fell within the range fixed by the European Pharmacopoeia. CONCLUSION: The method described is an effective way of preparing large quantities of IVIG (lyophilized and liquid) which fulfils the standard requirements of both the European Pharmacopoeia and the World Health Organization. PMID- 11438030 TI - Automated serological compatibility testing using a solid-phase test and standard laboratory equipment. AB - BACKGROUND AND OBJECTIVES: To prove the feasibility of a semi-automated cross match procedure using a commercially available solid-phase microplate test and standard laboratory equipment. MATERIAL AND METHODS: The new procedure was evaluated against the conventional spin tube technique and the gel centrifugation system. RESULTS: The sensitivity of the method and the rate of non-specific reactions were equal to those for the other test systems. The samples taken from the red cell concentrates for cross-matching remained stable for the shelf-life of the product. CONCLUSION: The semi-automated cross-match was successfully introduced in our routine laboratory as a means to process large numbers of tests. PMID- 11438031 TI - Direct evidence for the existence of Miltenberger antigen. AB - The Miltenberger (Mi) subsystem, which originally consisted of four phenotypes, now has 11 phenotypes. The antigens of this subsystem belong to the MNS blood group system. The Mia antigen has been reported to be present on red blood cells with several Miltenberger phenotypes, namely: Mi.I, Mi.II, Mi.III, Mi.IV, Mi.VI and Mi.X. However, the existence of the Mia antigen as a separate entity has been in question and difficult to prove with polyclonal reagents. We report the first monoclonal anti-Mia (GAMA210), whose epitope is TNDKHKRD or QTNDMHKR, and thereby confirm the existence of the Mia antigen. PMID- 11438032 TI - The ability of cellular assays to predict the necessity for cordocenteses in pregnancies at risk of haemolytic disease of the newborn. PMID- 11438033 TI - Naturally occurring anti-Fy(b) + C(w). PMID- 11438034 TI - B cells, BAFF/zTNF4, TACI, and systemic lupus erythematosus. AB - B cells and B-cell/T-cell collaborations are instrumental in the pathophysiology of systemic lupus erythematosus (SLE). This commentary highlights in particular the newly discovered role of B-cell-activating factor (BAFF; also known as TALL 1, THANK, BlyS, and zTNF4) as a positive regulator of B-cell functions, such as B cell activation and differentiation. Two members of the tumor necrosis factor(TNF)-receptor superfamily were recently identified as receptors for BAFF on B cells. The interaction between BAFF and its receptors may be important in the pathogenesis of lupus. Advances in our understanding of abnormalities in immune regulation in lupus might provide the opportunity to improve our current therapeutic approaches to this disorder. PMID- 11438035 TI - NF-kappa B in rheumatoid arthritis: a pivotal regulator of inflammation, hyperplasia, and tissue destruction. AB - The transcription factor NF-kappa B has been well recognized as a pivotal regulator of inflammation in rheumatoid arthritis (RA), but recent developments revealed a broad involvement of NF-kappa B in other aspects of RA pathology, including development of T helper 1 responses, activation, abnormal apoptosis and proliferation of RA fibroblast-like synovial cells, and differentiation and activation of bone resorbing activity of osteoclasts. In agreement with this, studies in animal models of RA have demonstrated the high therapeutic efficacy of specific inhibitors of NF-kappa B pathway, indicating the feasibility of anti-NF kappa B therapy for human disease. PMID- 11438036 TI - Links between complement deficiency and apoptosis. AB - Deficiency in the classical pathway complement components displays a hierarchical association with the development of systemic lupus erythematosus (SLE). In addition, SLE causes consumption of complement. C1q- and C4-deficient mice develop a lupus-like disease and exhibit impaired clearance of apoptotic cells. The autoantigens targeted in SLE have been localised to the surface of apoptotic cells, which may be the source of these antigens. Although apoptosis was originally thought to be an immunologically inert process, dendritic cells can present epitopes derived from apoptotic cells, and immunization with apoptotic cells leads to the generation of autoantibodies. These findings taken together indicate that a defect in complement-dependent clearance of apoptotic cells may increase susceptibility to the development of autoimmunity. PMID- 11438037 TI - A follow-up to "Anti-cytokine therapy in chronic destructive arthritis" by Wim B van den Berg. AB - In recent years, the effectiveness of anti-TNF therapy in treating rheumatoid arthritis (RA) has become apparent. While trials of IL-1 receptor antagonist in RA have been encouraging, it clearly is more difficult to target two molecules (IL-1 alpha and beta) than one (TNF-alpha). In his review article, Professor Wim van den Berg argues that both TNF-alpha and IL-1 must be blocked in RA and that although TNF is clearly a potent inflammatory molecule, the dominant cytokine in the subsequent degradation of the joint tissue is IL-1. This commentary discusses his hypothesis in light of animal studies and the limitations of the conclusions that can be drawn from them. More broadly, it discusses the biology of TNF-alpha and IL-1 and suggests explanations of why TNF-alpha is a pivotal cytokine in this disease. PMID- 11438039 TI - Genetic epidemiology. Approaches to the genetic analysis of rheumatoid arthritis. AB - The basis of susceptibility to rheumatoid arthritis (RA) is complex, comprising genetic and environmental susceptibility factors. We have reviewed the available approaches to the investigation of the genetic basis of complex diseases and how these are being applied to RA. Affected-sibling-pair methods for nonparametric linkage analysis, linkage-disequilibrium-based approaches, transmission disequilibrium testing, and disease-association studies are discussed. The pros, cons, and limitations of the approaches are considered and are illustrated by examples from the literature about rheumatoid arthritis. PMID- 11438040 TI - Bone loss. Therapeutic approaches for preventing bone loss in inflammatory arthritis. AB - Inflammatory arthritides are commonly characterized by localized and generalized bone loss. Localized bone loss in the form of joint erosions and periarticular osteopenia is a hallmark of rheumatoid arthritis, the prototype of inflammatory arthritis. Recent studies have highlighted the importance of receptor activator of nuclear factor-kappa B ligand (RANKL)-dependent osteoclast activation by inflammatory cells and subsequent bone loss. In this article, we review the pathogenesis of inflammatory bone loss and explore the possible therapeutic interventions to prevent it. PMID- 11438041 TI - 21st European Workshop for Rheumatology Research, Vienna, Austria, 1-4 March 2001. AB - Major advances in technology now drive how we approach questions in immunology, particularly in analyzing complex data sets commonly encountered in genomics and proteomics studies. Active areas of investigation include development of novel technologies, identification of elusive target antigens for RA and other diseases, dissection of signaling pathways connecting the lymphocyte cell surface with the nucleus, and exploration of new avenues for therapeutic interventions. The European Workshop for Rheumatology Research (EWRR) is a forum for many European and non-European scientists to present research findings of high quality. Arthritis researchers from around the globe should be strongly encouraged to attend future meetings, the next of which is the 22nd EWRR meeting in Leiden, the Netherlands, in 2002. PMID- 11438042 TI - Inflammatory microcrystals induce murine macrophage survival and DNA synthesis. AB - The interaction of particulates with resident macrophages is a consistent feature in certain forms of crystal-induced inflammation, for example, in synovial tissues, lung, and the peritoneum. The mitogenic activity of basic calcium phosphate (BCP) crystals and calcium pyrophosphate dihydrate (CPPD) crystals on synovial fibroblasts has been considered relevant to the synovial hyperplasia observed in crystal-induced arthritis. The aim of the study was to determine whether microcrystals such as these could enhance macrophage survival and induce DNA synthesis, thus indicating that they may contribute to the tissue hyperplasia. Murine bone-marrow-derived macrophages were treated in vitro with microcrystals, the cell numbers were monitored over time, and DNA synthesis was measured as the incorporation of [methyl-(3)H]thymidine (TdR). We report here that BCP, monosodium urate, talc, and, to a lesser extent, CPPD crystals promote macrophage survival and DNA synthesis; the latter response is particularly striking in the presence of low concentrations of macrophage-colony stimulating factor (M-CSF, CSF-1). Enhanced macrophage survival or proliferation may contribute to the synovial hyperplasia noted in crystal-associated arthropathies, as well as to talc-induced inflammation and granuloma formation. The crystals studied join the list of particulates having these effects on macrophages, indicating the generality of this type of response. PMID- 11438043 TI - Results of a phase-I/II randomized, masked, placebo-controlled trial of recombinant human interleukin-11 (rhIL-11) in the treatment of subjects with active rheumatoid arthritis. AB - Interleukin-11 (IL-11) is a pleiotropic cytokine that regulates the growth and development of hematopoietic stem cells and decreases the proinflammatory mediators of cytokine and nitric oxide production. In animal models of arthritis, treatment with recombinant human IL-11 (rhIL-11) reduces both the level of synovitis and the histologic lesion scores in the joints. The goal of this phase I/II study in adults with rheumatoid arthritis (RA) was to evaluate the safety and clinical activity of different doses and schedules of rhIL-11 in patients with active RA for whom treatment with at least one disease-modifying antirheumatic drug had failed. This was a multicenter, randomized, placebo controlled trial that evaluated the safety and tolerability of rhIL-11 in 91 patients with active RA. rhIL-11 was administered subcutaneously; patients were randomized into one of five treatment groups (ratio of rhIL-11 to placebo, 4:1). Patients were treated for 12 weeks with either 2.5 or 7.5 microg/kg of rhIL-11 or placebo twice per week or 5 or 15 microg/kg of rhIL-11 or placebo once per week. The status of each subject's disease activity in accordance with the American College of Rheumatology (ACR) criteria was assessed before, during, and after completion of administration of the study drug. Administration of rhIL-11 was well tolerated at all doses and schedules. The most frequent adverse event was a reaction at the injection site. The data suggest a statistically significant reduction in the number of tender joints (P < 0.008) at the 15 microg/kg once weekly dose schedule but showed no overall significant benefit at the ACR criterion of a 20% response. The trial showed rhIL-11 to be safe and well tolerated at a variety of doses and schedules over a 12-week treatment period in patients with active RA. The only adverse event clearly associated with rhIL-11 administration was reaction at the injection site. PMID- 11438044 TI - Autoimmune response to U1 small nuclear ribonucleoprotein (U1 snRNP) associated with cytomegalovirus infection. AB - The induction of autoantibodies to U1 small nuclear ribonucleoprotein (U1 snRNP) complexes is not well understood. We present evidence that healthy individuals with cytomegalovirus (CMV) infection have an increased frequency and quantity of antibodies to ribonucleoprotein, directed primarily against the U1-70k protein. A significant association between the presence of antibodies to CMV and antibodies to the total RNP targeted by the immune response to the spliceosome (to both the Sm and RNP; Sm/RNP) was found for patients with systemic lupus erythematosus (SLE) but not those with mixed connective-tissue disease. CMV thus may play a role in inducing autoimmune responses in a subset of patients with systemic lupus erythematosus. PMID- 11438045 TI - Approaches to enhancing the retroviral transduction of human synoviocytes. AB - This report concerns a clinical trial for rheumatoid arthritis (RA), approved by the US National Institutes of Health and the Food and Drug Administration. An amphotropic retrovirus (MFG-IRAP) was used ex vivo to transfer a cDNA encoding human interleukin-1 receptor antagonist (IL-1Ra) to synovium. The protocol required the transduced cells to secrete at least 30 ng IL-1Ra/10(6) cells per 48 h before reimplantation. Here we have evaluated various protocols for their efficiency in transducing cultures of human rheumatoid synoviocytes. The most reliably efficient methods used high titer retrovirus (approximately 10(8) infectious particles/ml). Transduction efficiency was increased further by exposing the cells to virus under flow-through conditions. The use of dioctadecylamidoglycylspermine (DOGS) as a polycation instead of Polybrene (hexadimethrine bromide) provided an additional small increment in efficiency. Under normal conditions of static transduction, standard titer, clinical grade retrovirus (approximately 5 x 10(5) infectious particles/ml) failed to achieve the expression levels required by the clinical trial. However, the shortfall could be remedied by increasing the time of transduction under static conditions, transducing under flow-through conditions, or transducing during centrifugation. PMID- 11438046 TI - Dry eye after laser in situ keratomileusis. AB - PURPOSE: To determine whether patients undergoing laser in situ keratomileusis have postoperative dry eye. METHODS: In this retrospective, interventional case series, 124 eyes of 64 consecutive patients who underwent laser in situ keratomileusis were examined for a dry eye symptom, Schirmer test with anesthesia, tear clearance rate, tear break-up time, vital staining for ocular surface, corneal sensitivity, and blink rate. All values were compared before and after surgery (1 month, 3 months, 6 months, and 1 year) and values of P <.05 were considered statistically significant. RESULTS: Subjective score for dryness was increased after laser in situ keratomileusis. Tear function index (Schirmer value/tear clearance rate) and break-up time were decreased until 1 month (tear function index) and 3 months (break-up time) after laser in situ keratomileusis but recovered to preoperative levels thereafter. Fluorescein and rose bengal scores were unchanged at all follow-up points except for improvement of the rose bengal score at 6 months. Corneal sensitivity was decreased at 1 month and 3 months, and returned to the preoperative level at 6 months after laser in situ keratomileusis. Blink rate was decreased at 3 months, 6 months, and 1 year after laser in situ keratomileusis. CONCLUSIONS: Patients undergoing laser in situ keratomileusis develop dry eye with compromised tear function for at least 1 month after surgery. Use of artificial tears in the early postoperative period may help to prevent unwanted symptoms and ocular surface damage. PMID- 11438047 TI - Treatment of recalcitrant recurrent corneal erosions with inhibitors of matrix metalloproteinase-9, doxycycline and corticosteroids. AB - PURPOSE: To review the efficacy of inhibitors of matrix metalloproteinase-9, corticosteroids, and doxycycline for treatment of recalcitrant recurrent corneal erosion. METHODS: Retrospective, clinic-based, interventional case series. The medical records of seven consecutive patients who were treated between January 1995 to January 2000 for recurrent corneal erosion who had not responded to conventional therapy were reviewed. Treatment of seven eyes of seven patients consisted of oral doxycycline (50 mg, two times a day) for 2 months along with a topical corticosteroid (either methylprednisolone 1%, prednisolone acetate 1%, or fluoromethalone 0.1%) three times a day, for 2 to 3 weeks. The effects of doxycycline and methylprednisolone on metalloproteinase-9 activity in human corneal epithelial cultures were evaluated by gelatin zymography and a commercial metalloproteinase-9 activity assay kit. RESULT: Fingernail injury in three of the seven eyes was the most common form of corneal injury. There was no evidence of epithelial basement membrane or corneal stromal dystrophy in any of the patients, although epithelial microcysts were observed in the involved area in three patients. One eye had intact elevated corneal epithelium that showed abnormal diffuse staining with fluorescein dye, and six eyes had a corneal epithelial defect at the time of presentation. In all seven eyes, pain resolved and epithelial defects healed within 2 to 10 days after initiation of therapy. No recurrence was observed during an average follow-up period of 21.9 months (range, 1.5 to 60 months). Methylprednisolone and doxycycline each produced a statistically significant decrease in amount and activity of metalloproteinase-9 in conditioned media of human corneal epithelial cultures. CONCLUSIONS: Therapy with a combination of medications that inhibit metalloproteinase-9 produced rapid resolution and prevented further recurrence of cases of recurrent corneal erosions that were unresponsive to conventional therapies. PMID- 11438048 TI - Tectonic sclerokeratoplasty and tectonic penetrating keratoplasty as treatment for perforated or predescemetal corneal ulcers. AB - PURPOSE: To report the clinical outcome of patients with perforated or predescemetal corneal ulcers treated by tectonic, centric or eccentric penetrating keratoplasty or by tectonic sclerokeratoplasty. DESIGN: Nonrandomized clinical trial. METHODS: The study included 60 patients (60 eyes) with perforated or predescemetal corneal ulcers who were consecutively operated on by the same surgeon. Fifty-two patients underwent tectonic penetrating centric or eccentric keratoplasty. Eight patients with paralimbal corneal ulcers underwent tectonic sclerokeratoplasty. A control group consisted of 76 patients (76 eyes) electively undergoing central penetrating keratoplasty for treatment of inactive central corneal scars. RESULTS: In the study group with perforated or predescemetal corneal ulcers, best-corrected postoperative visual acuity ranged from perception of light to 0.80 (median, 0.10), with 54 of 60 eyes (90%) attaining an improvement of best visual acuity. In 10 of 60 patients (16.7%), tectonic penetrating keratoplasty had to be re-performed because of a recurring corneal ulcer. Patients with sclerokeratoplasty and patients with eccentric keratoplasty did not differ in clinical outcome, despite larger trephine and corneal lesion size in the sclerokeratoplasty group. Among study patients compared with control patients, postoperative visual acuity was significantly lower (P =.01), postoperative refractive and keratometric astigmatism were significantly higher (P <.05), and immunologic graft reactions (P =.02) and suture loosening (P <.001) occurred significantly more often. CONCLUSIONS: Eyes with perforated corneal ulcers or predescemetal corneal ulcers can usually be saved by tectonic keratoplasty or sclerokeratoplasty, with a moderate to considerable amount of remaining useful vision. In case of doubt, one may prefer conservative treatment of corneal ulcers and to electively perform central keratoplasty when the ulcers have healed. For selected clinical situations, sclerokeratoplasty is an alternative to keratoplasty for surgical treatment of paralimbal corneal defects. PMID- 11438049 TI - Use of vitamin supplements and cataract: the Blue Mountains Eye Study. AB - PURPOSE: To investigate relationships between use of vitamin supplements and the three principal cataract types in a population-based sample. METHODS: We studied 2873 of the 3654 participants (79%) aged 49 to 97 years attending the cross sectional Blue Mountains Eye Study who completed a detailed food frequency questionnaire, which included type, dose, and duration of vitamin supplement use. Masked grading of nuclear, cortical, and posterior subcapsular opacities from lens photographs was performed, using the Wisconsin method. RESULTS: Use of multivitamin supplements was associated with reduced prevalence of nuclear cataract, odds ratio 0.6, 95% confidence interval 0.4 to 1.0, P =.05. For both nuclear and cortical cataract, longer duration of multivitamin use was associated with reduced cataract prevalence (nuclear cataract, trend P =.02; cortical cataract, trend P =.03). Use of thiamin supplements was associated with reduced prevalence of nuclear (odds ratio 0.6, confidence interval 0.4 to 1.0, P =.03, dose trend P =.03) and cortical cataract (odds ratio 0.7, confidence interval 0.5 to 0.9, P =.01, dose trend P =.02). Riboflavin (odds ratio 0.8, confidence interval 0.6 to 1.0, P =.05) and niacin (odds ratio 0.7, confidence interval 0.6 to 1.0, P =.04) supplements exerted a weaker protective influence on cortical cataract. Vitamin A supplements were protective against nuclear cataract (odds ratio 0.4, confidence interval 0.2 to 0.8, P =.01, dose trend P =.01). Folate (odds ratio 0.4, confidence interval 0.2 to 0.9, P =.03) appeared protective for nuclear cataract, whereas both folate (odds ratio 0.6, confidence interval 0.3 to 0.9, P =.01, dose trend P =.04) and vitamin B12 supplements (odds ratio 0.7, confidence interval 0.5 to 1.0, P =.03, dose trend P =.02) were strongly protective against cortical cataract. CONCLUSIONS: Long-term use of multivitamins, B group and vitamin A supplements was associated with reduced prevalence of either nuclear or cortical cataract. A strong protective influence on cortical cataract, from use of folate or vitamin B12 supplements, is a new finding. PMID- 11438050 TI - The long-term outcome of glaucoma filtration surgery. AB - PURPOSE: To determine the long-term outcome of glaucoma filtration surgery in preserving vision. Visual loss from progressive glaucomatous damage and from complications of surgery, both short and long term, were included. METHODS: A retrospective, community-based, longitudinal study of residents of Olmsted County, Minnesota, who were newly diagnosed with open-angle glaucoma between 1965 and 1980 and underwent filtration surgery in these or subsequent years through 1998. Intraocular pressure (IOP), visual acuity, visual fields, and progression to legal blindness were monitored. Kaplan-Meier analysis was used to determine the cumulative probabilities of changes in these parameters. RESULTS: 73 eyes of 49 patients underwent conventional filtration surgery. Analysis of the first eye having surgery revealed a mean preoperative IOP of 27.6 +/- 8.5 mm dropping to 16.7 +/- 5.6 mm at year one, and remaining in this range throughout follow-up (14.7 +/- 3.0 mm at 10 years; with or without use of medications). The probability of progression to blindness was 46% at 10 years after surgery, as calculated by Kaplan-Meier analysis. Eyes going blind had a postoperative IOP equal to or lower than those not becoming blind (14.0 +/- 4.4 vs. 15.4 +/- 3.0 at postoperative year 10). Eyes going blind had more advanced field loss at the time of surgery, with scotomas above and below the horizontal axis, than eyes not going blind, which had scotomas in only one hemifield. Three patients developed late bleb leaks; two patients developed endophthalmitis. The probability of undergoing cataract surgery was 37% by 10 years postoperatively, which did not differ significantly from the cohort of patients not undergoing surgery at a comparable time point. CONCLUSIONS: Filtration surgery was associated with a 54% probability of preservation of vision from progression to legal blindness at 10 years after surgery. Patients becoming blind had more advanced field loss at the time of surgery; IOP was similar between those going blind and those retaining vision. PMID- 11438051 TI - Autologous blood injection for late-onset filtering bleb leak. AB - PURPOSE: To report the outcomes of autologous blood injections for late-onset filtering bleb leak. METHODS: Retrospective chart review of all eyes that had autologous blood injection(s) for filtering bleb leak occurring at least 2 months after trabeculectomy at the Indiana University Medical Center. Successful treatment was defined as resolution of the bleb leak and no need for additional glaucoma medications. Failure was defined as a persistent bleb leak, intraocular pressure greater than 21 mm Hg, or the occurrence of a vision-threatening event related to the procedure. RESULTS: Thirty-two eyes of 31 patients had autologous blood injection for filtering bleb leak and were followed for a mean of 4.9 months (SD, 9.2; range, 1 to 37 months). Twenty-three eyes (72%) were outright failures because of persistence of the leak. Nine eyes (28%) had an initially successful outcome, but the success rate decreased over time as bleb leaks recurred in three of the nine eyes at 5, 6, and 37 months. No patient characteristics correlated with outcome. Mean intraocular pressure increased from pretreatment to final examination (4.5 to 6.5 mm Hg, P =.003). Mean logarithm of minimal angle of resolution (logMAR) vision remained unchanged from pretreatment to final examination (P =.55). Blood seepage into the anterior chamber after autologous blood injection was common but transient. CONCLUSIONS: Autologous blood injection is of limited success in treating late-onset filtering bleb leak. PMID- 11438052 TI - Influence of cataract surgery on automated perimetry in patients with glaucoma. AB - PURPOSE: To investigate the influence of cataract surgery on automated perimetry in patients with glaucoma. DESIGN: Interventional case series. METHODS: A total of 105 eyes of 105 consecutive patients with glaucoma who were scheduled for cataract surgery underwent Humphrey static threshold testing (30-2 program) before and at 1 month after surgery. These eyes were divided into two groups based on the presence of absolute or near absolute (dense) scotomata before surgery (minimum threshold value 5 decibels or less). Changes in numbers of the dense scotomata, mean deviation, pattern standard deviation, and corrected pattern standard deviation were analyzed. RESULTS: No significant changes were found in the numbers of dense scotomata and central dense scotomata within five degree visual fields between before and after surgery. In the group with dense scotomata, the mean pattern standard deviation and corrected pattern standard deviation worsened significantly after surgery, whereas the mean deviation improved significantly. However, in the group without dense scotomata, the pattern standard deviation and corrected pattern standard deviation showed a slight improvement, whereas the mean deviation improved significantly. When the central scotoma was present in two or fewer meridians before surgery, the mean visual acuity after surgery reached 20/25 and was better than that in patients with three or four central scotomata (P =.0014). CONCLUSIONS: Cataract does not produce a dense scotoma on automated perimetry. However, because it does produce relative scotomata, actual glaucomatous visual field defects may be hidden to some extent. When a central dense scotoma is present before surgery in two or fewer meridians, the patient may well achieve substantial improvement in postoperative visual acuity. PMID- 11438053 TI - Rate of visual field loss and long-term visual outcome in primary open-angle glaucoma. AB - PURPOSE: To evaluate long-term visual field outcome in primary open-angle glaucoma. METHODS: In this retrospective cohort study, 40 eyes of 40 patients with primary open-angle glaucoma with elevated intraocular pressure and a minimum of 8-year longitudinal series of visual fields were plotted with Goldmann perimeter. Eyes with any other ocular disease except cataract were excluded. Manual grid templates were used to quantify the visual fields. Linear regression was performed to estimate the rate of visual field decline. Pertinent clinical factors were evaluated for statistical association with the rate of decline. Long term clinical outcome including visual acuity, rate of legal blindness, and rate of medical and surgical interventions was also measured. RESULTS: In the 40 eyes studied, with a mean follow-up of 14 years, the visual field score decreased at the rate of -1.5% per year. Overall, 68% showed significant decrease, and the rate of decrease among these eyes was -2.1% per year. Five eyes became legally blind from glaucoma; the cumulative rate of blindness from glaucoma was 19% at 22 years. Higher intraocular pressure and greater number of antiglaucoma medications on initial presentation were associated with faster and slower deterioration of visual field (compared with the average), respectively. CONCLUSIONS: With standard glaucoma therapy, the rate of visual field loss in primary open-angle glaucoma is slow. Lower intraocular pressure and more antiglaucoma medications are associated with slower visual field decline. Legal blindness from glaucoma is 19% over a follow-up of 22 years. PMID- 11438054 TI - Interobserver variability of optic disk variables measured by confocal scanning laser tomography. AB - PURPOSE: To assess the interobserver variation of confocal laser scanning tomographic measurements of the optic nerve head and to address the question of whether the addition of clinical optic disk photographs is helpful in outlining the optic disk margin and in reducing the observer-related variation of the measurements. PATIENTS AND METHODS: Optic disk variables for 16 eyes of 16 patients with glaucoma, generated by confocal laser scanning laser tomography (Heidelberg Retina Tomograph), were independently evaluated by four experienced glaucoma specialists, and the interobserver variability was calculated. A second separate review by the same observers included the use of clinical stereoscopic color optic nerve head photographs to aid definition of the optic disk margin. RESULTS: Optic disk parameters with the smallest interobserver variation were cup shape measure, maximum cup depth, height variation contour, and mean height contour. The intraobserver variation of these parameters did not increase when clinical optic disk slides were additionally available. Parameters with the highest interobserver variation were volume below surface, volume below reference, volume above surface, and volume above reference. The observer variation of these optic disk parameters increased significantly for two of the four examiners when clinical optic disk slides were additionally available for outlining the optic disk margin. CONCLUSION: Confocal laser scanning tomography of the optic nerve head can be improved significantly if clinical optic disk photographs are additionally available to help in outlining the optic disk margin. Because interobserver variation in the tomographic optic disk measurements can be significant, even if experienced observer are involved, tomographic optic disk measurements may be centralized in reading centers in the case of multicenter studies. PMID- 11438055 TI - Optic nerve blood flow is diminished in eyes of primary open-angle glaucoma suspects. AB - PURPOSE: The purpose of this study was to evaluate optic nerve blood flow in primary open-angle glaucoma suspect eyes with normal automated visual fields, in an attempt to elucidate how early in the glaucomatous disease process changes in optic nerve blood flow become apparent. METHODS: Twenty-one eyes (21 patients) suspected of having primary open-angle glaucoma were studied prospectively and compared with a previously reported cohort of 22 eyes (22 patients) with primary open-angle glaucoma and 15 eyes (15 subjects) of age-matched controls. Primary open-angle glaucoma suspect eyes had untreated intraocular pressure greater than 21 mm Hg and normal visual fields using Humphrey program 24-2 or 30-2 with a full threshold strategy. Laser Doppler flowmetry was used to measure optic nerve head blood velocity, volume, and flow at four quadrants in the optic nerve, in the cup, and in the foveola of one eye of each patient. The mean flow from the superotemporal rim, inferotemporal rim, and cup was calculated (Flow(3)) and identified as the main outcome measure. Measurements from primary open-angle glaucoma suspect eyes were compared with corresponding measurements from controls and eyes with primary open-angle glaucoma; a Student t test was employed with a Bonferroni corrected P value of.025 to account for comparisons of primary open angle glaucoma suspects both to controls and to eyes with primary open-angle glaucoma. RESULTS: Compared with controls, Flow(3) was 24% lower in primary open angle glaucoma suspect eyes (P <.0003). In primary open-angle glaucoma suspect eyes, flow was 16% lower in the superotemporal rim (P <.007), 35% lower in the cup (P <.007), and 22% lower in the inferotemporal neuroretinal rim (P <.029) compared with controls. No significant difference between primary open-angle glaucoma suspect and control eyes was seen in the inferonasal rim, superonasal rim, or foveola. No significant difference was detected at any location between primary open-angle glaucoma suspect eyes and eyes with primary open-angle glaucoma. CONCLUSIONS: Laser Doppler flowmetry detected circulatory abnormalities in primary open-angle glaucoma suspects who did not have any manifest visual field defect. Decreases in flow in glaucoma suspects were similar in magnitude to those of subjects with primary open-angle glaucoma. These data suggest that impaired optic nerve blood flow develops early in the glaucomatous process and does not develop solely as a result of glaucoma damage. PMID- 11438056 TI - Asteroid hyalosis in a population: the Beaver Dam eye study. AB - PURPOSE: To estimate the prevalence of asteroid hyalosis and to examine correlates of asteroid hyalosis in a population-based cohort. METHODS: The population of Beaver Dam, Wisconsin, that was 43 to 86 years of age was examined from 1988 to 1990 (n = 4926). The population is predominantly white (99.4%) northern European. Asteroid hyalosis was determined from stereoscopic fundus photographs of three standard fields. RESULTS: Fundus photographs were gradable in 4747 subjects. Asteroid hyalosis was present in 1.2% (95% confidence interval, 0.9,1.5%). In subjects in which it was present, asteroid hyalosis was bilateral in 9%. Prevalence increased significantly (P <.001) with age from 0.2% in subjects 43 to 54 years to 2.9% in subjects 75 to 86 years. After adjusting for age, men were more likely (1.8%) to have asteroid hyalosis than women (0.8%). After adjusting for age and sex, asteroid hyalosis was significantly more likely to be found in subjects with greater body mass (P =.02) and higher alcohol consumption (P =.03). There were nonsignificant trends with systolic blood pressure (P =.07), serum cholesterol (P =.09), and serum albumin (P =.09). It was not significantly associated (P >.05) with diastolic blood pressure, hypertension, diabetes, cardiovascular disease history, high-density-lipoprotein cholesterol, serum calcium, cigarette smoking, physical activity, intraocular pressure, or refractive error. These relationships were confirmed in a multivariable logistic model. CONCLUSIONS: The current study documents the infrequency of asteroid hyalosis in the population as graded from three photographic fields of the fundus. It does not support previous observations of an association of asteroid hyalosis with diabetes or refractive error. The relevance of the new associations reported (body mass, alcohol) remains to be determined. PMID- 11438057 TI - Randomized comparison of diode laser photocoagulation versus cryotherapy for threshold retinopathy of prematurity: seven-year outcome. AB - PURPOSE: To report the structural and functional outcomes at a minimum of 7 years postmenstrual age after randomized treatment of threshold retinopathy of prematurity with laser ablation or cryotherapy. METHODS: Nineteen patients were entered into a prospective, randomized protocol, in which one eye received cryotherapy, while the other eye received diode laser photocoagulation. Asymmetric eyes were randomly assigned. Two patients have died, and seven were no longer available for 7-year outcome examinations, leaving 10 children for analysis. RESULTS: Six males and four females with a mean birthweight of 631 g and a mean gestational age of 24.8 weeks were examined. Eight were symmetrical cases and treated in both eyes. Of these, there were six concordant and two discordant structural outcomes. The laser-treated eyes had the favorable outcome in each instance. The geometric mean visual acuity of the paired eyes after laser photocoagulation was 20/33, and after cryotherapy it was 20/133 (P =.03). The mean refractive error was -6.50 diopters after laser photocoagulation and -8.25 diopters after cryotherapy (P =.27), although one of the cryotherapy eyes could not be refracted because of phthisis. CONCLUSIONS: Laser photocoagulation appears to be associated with a structural and functional outcome at least as good as cryotherapy 7 years after therapy. Visual acuity and refractive error data suggest that laser photocoagulation may have an advantage over cryotherapy. PMID- 11438058 TI - Optical coherence tomography of tractional macular elevations in eyes with proliferative diabetic retinopathy. AB - PURPOSE: To document the findings of optical coherence tomography in eyes with tractional macular elevation associated with proliferative diabetic retinopathy, paying special attention to differentiating between tractional retinal detachment and retinoschisis. DESIGN: Observational case series. METHODS: We retrospectively reviewed images of optical coherence tomography obtained from 17 consecutive eyes (17 patients) with proliferative diabetic retinopathy without vitreous opacity and showing tractional macular elevation with or without foveal involvement. RESULTS: Tractional retinal detachment and retinoschisis could be differentiated in the cross-sectional images of optical coherence tomography. Retinoschisis with or without associated retinal detachment was observed in 16 of 17 eyes (94%), whereas retinal detachment was observed in only six of 17 eyes (35%). Five eyes showed both findings of retinal detachment and retinoschisis. CONCLUSIONS: Optical coherence tomography is useful for distinguishing tractional retinoschisis from retinal detachment in eyes with proliferative diabetic retinopathy and macular elevation. Tractional retinoschisis with or without retinal detachment is the most frequent pattern of tractional macular elevation in eyes with proliferative diabetic retinopathy. PMID- 11438059 TI - Theoretical investigation of the role of choriocapillaris blood flow in treatment of subfoveal choroidal neovascularization associated with age-related macular degeneration. AB - PURPOSE: To investigate the relationship between choriocapillaris blood flow and blood flow through an overlying choroidal neovascularization, as it relates to photocoagulation-induced changes in the choriocapillaris circulation. METHODS: A theoretical model that simulates the blood flow in the choriocapillaris and choroidal neovascularization of the human eye was developed, based on histologically determined vascular geometry and experimentally measured blood pressure gradients. The choriocapillaris blood pressure and blood flow were examined before and after simulated photocoagulation of various Sattler layer vessels entering the choriocapillaris in the vicinity of the choroidal neovascularization. (The Sattler layer is the inner layer of medium-sized choroidal vessels that includes both arterioles and venules that supply the choriocapillaris.) RESULTS: The theoretical model showed that both partial and complete occlusion of either Sattler arteriole or venous vessels in the vicinity of the capillary-like vessels connecting a choroidal neovascularization to the underlying choriocapillaris results in significant choroidal neovascularization blood flow reduction. These theoretical results are similar to clinically observed changes induced by laser photocoagulation of feeder vessels. (In this discussion, the term "feeder vessels" refers to those vessels in an indocyanine green angiogram image that appear to supply blood to a choroidal neovascularization; these vessels appear to be Sattler layer vessels, rather than the histologically demonstrated short, capillary-like vessels that form choriocapillaris-choroidal neovascularization communications.) CONCLUSIONS: Reduction of choriocapillaris blood flow underlying a choroidal neovascularization may be sufficient to reduce the blood flow rate in the choroidal neovascularization and thereby reduce the associated retinal edema. The results also suggest that reduction of choriocapillaris blood flow may be the common hemodynamic event associated with the successful application of several currently practiced methods of choroidal neovascularization treatment, including feeder vessel photocoagulation, photodynamic therapy, transpupillary thermotherapy, and prophylactic drusen photocoagulation. Ultimately, this model may be useful in determining optimal placement of laser photocoagulation burns to achieve a desirable perturbation in choroidal blood flow distribution and thereby reduce choroidal neovascularization blood flow to the extent necessary to obliterate associated retinal edema. PMID- 11438060 TI - Basic fibroblast growth factor inhibits choriocapillaris atrophy in rabbit. AB - PURPOSE: To examine the effect of basic fibroblast growth factor on induced choriocapillaris atrophy in vivo. METHODS: Choriocapillaris atrophy was surgically induced in rabbits by a hydraulic retinal detachment followed by debridement of the retinal pigment epithelium under the detached retina. Three concentrations of basic fibroblast growth factor (0.1 microg/0.1 ml, 1 microg/0.1 ml, or 5 microg/0.1 ml) were injected into the subretinal space and into the vitreous cavity 1, 3, and 5 days after the surgery. For control, only Tris buffer was injected in the same manner. The rabbits were euthanized 7 days after the surgery. Choroidal vascular casts were made and examined by scanning electron microscopy. The choriocapillaris atrophy was quantified by computer-assisted image analysis of photographs of the choriocapillaries. The area of the choriocapillaris and number of intercapillary spaces in the choriocapillaris that corresponded to the density of the capillary network were measured. RESULTS: The average area of the choriocapillaris in the eyes treated with 1 microg/0.1 ml of basic fibroblast growth factor was significantly larger at 75.1 +/- 3.0% than that in the control eyes at 67.2 +/- 5.6% (P =.021). The average area of the choriocapillaris in the 0.1 microg/0.1 ml of basic fibroblast growth factor group was not statistically different from the control. The number of intercapillary spaces of the choriocapillaris was 132 +/- 12 in the 0.1 microg/0.1 ml of basic fibroblast growth factor group, 124 +/- 46 in the 1 microg/0.1 ml of basic fibroblast growth factor group, and 75 +/- 14 in the control group. The higher number of spaces in the treated group was statistically significant (P =.026). CONCLUSIONS: Basic fibroblast growth factor decreased the atrophy of the choriocapillaris after removal of the retinal pigment epithelium in rabbit eyes. These results suggest that basic fibroblast growth factor may play a role in the survival of the choriocapillaris in vivo. PMID- 11438061 TI - Is there a "lost" rectus muscle in strabismus surgery? PMID- 11438062 TI - Pigmented conjunctival squamous cell carcinoma simulating a conjunctival melanoma. AB - PURPOSE: To report a pigmented conjunctival squamous cell carcinoma that clinically simulated a conjunctival melanoma. DESIGN: Interventional case report. METHODS: Ocular examination, surgical excision, and clinicopathologic correlation. RESULTS: A 78-year-old white man developed a lightly pigmented mass at the temporal limbus of his right eye. The differential diagnosis included pigmented squamous cell carcinoma and malignant melanoma. Histopathologic examination revealed a malignant squamous cell tumor that contained foci of melanin pigment. The final diagnosis was pigmented conjunctival squamous cell carcinoma. CONCLUSION: Conjunctival squamous cell carcinoma may rarely be pigmented and simulate a conjunctival melanoma. PMID- 11438063 TI - Bilateral neurotrophic keratopathy complicating Vidian neurectomy. AB - PURPOSE: To present a case of bilateral neurotrophic keratopathy with miosis after bilateral Vidian neurectomy for vasomotor rhinitis. METHODS: Observational case report. RESULTS: A 68 year-old man presented with irritation and blurred vision in both eyes 2 weeks after bilateral Vidian neurectomy. Slit-lamp examination revealed a large epithelial defect, typical of neurotrophic keratopathy, in the inferior two-thirds of cornea in both eyes. Corneal sensitivity test with a Cochet-Bonnet anesthesiometer and electrical study of the blink reflex indicated bilateral trigeminal dysfunction. Both pupils were miotic, and tests with hydroxyamphetamine 1% and epinephrine 0.1% showed postganglionic sympathetic nerve damage. The corneal epithelial defects healed after 2 months of treatment with systemic prednisolone, vitamin B(12) and tarsorrhaphy. CONCLUSION: Neurotrophic keratopathy associated with dry eye syndrome may be a serious complication of Vidian neurectomy. PMID- 11438064 TI - Anterior capsular contraction after cataract surgery in vitrectomized eyes. AB - PURPOSE: To assess the contraction of continuous curvilinear capsulorhexis after cataract surgery in eyes with past pars plana vitrectomy. METHODS: In a prospective study, 16 eyes of 16 patients underwent phacoemulsification and implantation of a foldable acrylic intraocular lens after pars plana vitrectomy. Eyes after intensive or repeated vitrectomy were not included. Twenty eyes of 19 patients served as age-matched controls. Aqueous flare intensity was measured using the laser flare-cell meter 1 year after surgery. The area of anterior capsular opening (ACO) was determined by diaphanoscopy using the anterior eye segment analysis system EAS-1000 at 1 day and 1 year postoperatively. RESULTS: There was no significant difference in the mean ACO area between the vitrectomy and control groups both at 1 day and 1 year postoperatively. Aqueous flare intensity 1 year after surgery was slightly higher in the vitrectomy group, but the difference was not statistically significant. CONCLUSION: Eyes after simple vitrectomy are not at a higher risk of ACO contraction following cataract surgery. PMID- 11438065 TI - Capsular bag distention syndrome after combined cataract-lens implant surgery and Ahmed valve implantation. AB - PURPOSE: To describe the capsular bag distention syndrome after combined cataract extraction with posterior lens implant and aqueous drainage device. METHODS: Case report. RESULTS: A persistently shallow anterior chamber and low intraocular pressure developed after combined cataract extraction with posterior chamber lens implant and Ahmed aqueous drainage device. An optically empty space between the lens implant and posterior capsule was detected 18 days after surgery. The anterior chamber deepened within minutes after Nd:YAG posterior capsulotomy. CONCLUSION: The capsular bag distention syndrome needs to be included in the differential diagnosis of shallow anterior chamber with low intraocular pressure after combined cataract extraction and glaucoma valve implant surgery. PMID- 11438066 TI - Rhino-orbital mucormycosis in a patient with acquired immunodeficiency syndrome (AIDS) and neutropenia. AB - PURPOSE: To present a case of rhino-orbital mucormycosis in a patient with AIDS and neutropenia managed without exenteration. METHODS: Case report. RESULTS: A 60 year-old African-American man with AIDS developed neutropenia that was probably secondary to antiretroviral therapy. He developed right rhino-orbital mucormycosis and was treated with right partial ethmoidectomy with debridement and liposomal amphotericin B. The infection was cured without need for orbital exenteration, although visual acuity in his right eye ultimately was no light perception. CONCLUSION: Rhino-orbital mucormycosis is uncommon in patients with AIDS. When rhino-orbital mucormycosis occurs, patients require a careful search for an underlying metabolic derangement such as neutropenia. Treatment should be aggressive, but these patients may not require orbital exenteration. PMID- 11438067 TI - Presumed topiramate-induced bilateral acute angle-closure glaucoma. AB - PURPOSE: We describe a case of bilateral angle-closure glaucoma associated with oral topiramate therapy. METHODS: Interventional case report. Case report with echographic illustration. RESULTS: A 51-year-old man developed bilateral acute angle-closure glaucoma 2 weeks after beginning topiramate therapy for bipolar affective disorder. Laser peripheral iridotomy was performed in the right eye without resolution of the acute attack. Echography revealed lens thickening and ciliochoroidal detachments in both eyes. Visual acuity, intraocular pressure, and anterior and posterior segment anatomy normalized 2 weeks after cessation of topiramate therapy. CONCLUSION: Topiramate, a new sulfa-derivative antiepileptic medication, may cause idiosyncratic ciliochoroidal detachments and ciliary body edema leading to anterior displacement of the lens-iris diaphragm, lens thickening, and acute angle-closure glaucoma. PMID- 11438069 TI - Post-traumatic endophthalmitis involving Clostridium tetani and Bacillus spp. AB - PURPOSE: To report a case of post-traumatic infectious endophthalmitis caused by Clostridium tetani and Bacillus spp. METHODS: Case report. RESULTS: A 25-year-old man developed endophthalmitis after a traumatic corneoscleral laceration of his right eye by a concrete reinforcement bar. He underwent pars plana lensectomy and vitrectomy with aspiration of vitreous fluid and a conjunctival swab for cultures. Cultures from the conjunctival swab were negative for organisms. Cultures of the vitreous aspirate were positive for Bacillus species and C. tetani. He had received a tetanus toxoid booster at the emergency department. By the time the culture results became available, he had developed severe eye pain associated with marked orbital congestion, increased swelling and erythema of the lids, marked injection and chemosis of the conjunctiva, and subsequently underwent evisceration. The inflammation resolved after evisceration of the right eye, and he was discharged to home on doxycycline 100 mg orally two times daily for 10 days. CONCLUSION: We are unaware of previous reports of endophthalmitis involving C tetani and could find none in a computerized MEDLINE search. Patients with penetrating eye injury should be assessed for tetanus immunization status, and early intervention with tetanus toxoid booster and/or tetanus immune globulin should be considered if cultures are positive. PMID- 11438068 TI - Herpes simplex virus dermatitis in patients using latanoprost. AB - PURPOSE: To describe the possible association of latanoprost with herpetic dermatitis of the periocular skin. METHOD: Interventional case reports. A 79-year old woman with open-angle glaucoma developed a vesicular dermatitis of the left lower eyelid 14 months after starting latanoprost therapy. An 84-year-old man with pigmentary glaucoma developed a vesicular dermatitis of the right upper lid after 2 months of treatment with latanoprost and 8 days of treatment with tobramycin/dexamethasone for presumed bacterial conjunctivitis. In both cases, the dermatitis was characteristic of a herpetic infection. RESULTS: Latanoprost was discontinued in both cases. The woman was treated with vidarabine 3% ointment, and the man was not treated with antiviral agents. In both patients, the dermatitis healed uneventfully. The lesions of the man were cultured, and a biopsy was performed; herpes simplex virus type 1 was recovered from the culture and confirmed by immunofluorescence testing. CONCLUSION: Latanoprost, which has been associated with reactivation of herpetic keratitis, may also cause reactivation of herpetic dermatitis of the periocular skin. PMID- 11438070 TI - Progressive outer retinal necrosis syndrome in a lymphoma patient with good visual outcome. AB - PURPOSE: To report an HIV-negative lymphoma patient who developed progressive outer retinal necrosis syndrome and who had a good visual outcome after treatment with two-drug antiviral therapy and intravenous immunoglobulin. METHODS: Case report. RESULTS: A 43-year-old man with small lymphocytic lymphoma was diagnosed with progressive outer retinal necrosis in his left eye. Treatment was initiated with intravenous foscarnet and ganciclovir as well as intravenous gammaglobulin at a dose of 0.5 gm/kg per day for 5 days. On the second hospital day he was started on decadron 4 mg orally four times daily. No further posterior retinitis progression was observed despite severe immunosuppression. Visual acuity remained stable at 20/30 with 10 months' follow-up. CONCLUSIONS: The benefit of using gammaglobulin in progressive outer retinal necrosis is unknown. Given the rapid improvement seen in this patient's retinitis, it may be reasonable to consider the use of gammaglobulin in other cases of infectious retinitis in immunocompromised patients. PMID- 11438071 TI - Optical coherence tomography of bleb-like subretinal lesions after retinal reattachment surgery. AB - PURPOSE: To report the optical coherence tomographic characteristics of persistent bleb-like subretinal lesions after retinal reattachment surgery, including scleral buckling and cryopexy. METHODS: Case reports. RESULTS: In two eyes of two patients yellow to yellowish-orange bleb-like lesions were found in the posterior pole after retinal reattachment surgery. The common aspects of the surgery were scleral buckling procedures and cryopexy. There were subretinal precipitates in some of the lesions. The lesions showed no associated leakage during fluorescein or indocyanine green angiography. Optical coherence tomography demonstrated that the bleb-like lesions were tiny retinal detachments. CONCLUSION: Optical coherence tomography examination of bleb-like subretinal lesions showed that they were actually retinal detachments. Our patients and previously reported patients developed these lesions after scleral buckling and cryopexy. These lesions may represent persistent retinal detachment because of the presence of protein in the subretinal fluid. PMID- 11438072 TI - Postsurgical evaluation of idiopathic vitreomacular traction syndrome by optical coherence tomography. AB - PURPOSE: To report a case of idiopathic vitreomacular traction syndrome with preoperative and postoperative evaluation by optical coherence tomography. DESIGN: Interventional case report. METHODS: A 62-year-old woman presented with blurred vision in the left eye because of idiopathic vitreomacular traction syndrome, and she underwent a pars plana vitrectomy. Optical coherence tomography was performed before and after surgery. RESULTS: Preoperative optical coherence tomography, right eye, revealed residual adhesion of incomplete posterior vitreous detachment and edematous, thickened outer retina in the macula. A successful vitrectomy relieved vitreoretinal traction with nearly complete resolution of cystoid macular edema within 1 month after surgery, followed in subsequent months by gradual foveal depression resembling a lamellar macular hole. Resolution of subretinal serous fluid was delayed with complete disappearance, some 12 months after surgery, which correlated with a gradual improvement in visual acuity. CONCLUSION: Optical coherence tomography provides a sensitive anatomical evaluation of vitreomacular traction syndrome. Reorganization of retinal tissue after surgical intervention for vitreoretinal traction may be slower than is apparent from conventional examinations. PMID- 11438073 TI - Effective nasal limited macular translocation. AB - PURPOSE: To describe a case of effective foveal displacement toward the optic disk (nasal limited macular translocation) in a patient with a large subfoveal choroidal neovascularization secondary to age-related macular degeneration. METHODS: Case report. RESULTS: A 77-year-old white man presented with decreased vision of 20/400 due to subfoveal predominantly occult CNV secondary to age related macular degeneration in the left eye. The CNV, measuring 9 Macular Photocoagulation Study disk areas in size, was centered temporally relative to the fovea with a minimum desired translocation of 650 microm for nasal macular translocation. The patient underwent nasal LMT with punctate retinotomy and temporal chorioscleral infolding, followed by postoperative head-positioning on his right side. Effective LMT was achieved with a postoperative nasal foveal displacement of 1400 microm. The entire CNV was ablated with laser photocoagulation postoperatively. His vision improved to 20/40 6 months postoperatively. CONCLUSION: Nasal LMT is feasible and may be considered in patients with subfoveal CNV centered temporally relative to the fovea. PMID- 11438074 TI - Acute idiopathic blind spot enlargement syndrome: prolonged retinal dysfunction revealed by multifocal electroretinogram technique. AB - PURPOSE: To report the multifocal electroretinographic findings of a patient with acute idiopathic blind spot enlargement syndrome recorded at 2 weeks and 4 months after the onset. METHODS: A 35-year-old patient underwent static visual field (Humphrey, 30-2) and multifocal electroretinogram testing at 2 weeks and 4 months after the onset of acute idiopathic blind spot enlargement. The stimulus array for the multifocal electroretinogram consisted of 61 hexagons, and the total recording time was about 4 minutes. RESULTS: At 2 weeks, the patient had a large, well-demarcated scotoma centered on the blind spot, and its margin extended to within 5 degrees of fixation. The multifocal electroretinograms were depressed in the areas corresponding to the scotoma. At 4 months, her visual symptoms disappeared and static visual fields showed normal sensitivities at almost all locations. However, the multifocal electroretinograms still revealed reduced focal responses in a broad area around the blind spot. CONCLUSION: These results suggest that multifocal electroretinograms may be used to detect subclinical retinal dysfunction in patients with acute idiopathic blind spot enlargement. PMID- 11438075 TI - The relation of socioeconomic factors to the incidence of early age-related maculopathy: the Beaver Dam eye study. AB - PURPOSE: To investigate the relationship between socioeconomic factors and the 5 year incidence of age-related maculopathy. METHODS: The Beaver Dam Eye Study, a population-based cohort study, examined 3681 adults (range, 43-86 years of age at baseline) living in Beaver Dam, Wisconsin, at baseline and 5 years later. Standardized protocols for physical examination, including administration of a questionnaire that included questions regarding income, education level, and status and type of employment, and fundus photography to determine age-related maculopathy, were performed. RESULTS: While controlling for age and sex, less education, and being in a service-related occupation compared with a white collar professional occupation, was associated (P <.05) with the incidence of early age related maculopathy. CONCLUSION: These data show an association of education and occupation but not income with the incidence of early age-related maculopathy that appears independent of smoking or vitamin supplement use status. PMID- 11438076 TI - Combined hamartoma of the retina and retinal pigment epithelium associated with juvenile nasopharyngeal angiofibroma. AB - PURPOSE: To report a patient who had concomitant combined hamartoma of the retinal pigment epithelium and retina and juvenile nasopharyngeal angiofibroma. METHOD: Case report. RESULTS: An 18-year-old man presented with a juvenile nasopharyngeal angiofibroma and a combined hamartoma of the retina and retinal pigment epithelium in the ipsilateral eye. The diagnosis of juvenile nasopharyngeal angiofibroma was confirmed by histopathology, and the diagnosis of combined hamartoma was established by typical ophthalmoscopic and fluorescein angiographic findings. CONCLUSION: Juvenile nasopharyngeal angiofibroma may be associated with combined hamartoma of the retina and retinal pigment epithelium. PMID- 11438077 TI - Intermittent visual loss and exophthalmos due to the Blue rubber bleb nevus syndrome. AB - PURPOSE: To describe the occurrence of intermittent visual loss in a patient with orbital hemangioma due to the blue rubber bleb nevus syndrome. METHODS: Case report. RESULTS: A 70-year-old woman complained of intermittent blurred vision combined with pressure behind her left eye occurring after head bending or Valsalva maneuvers. Examination revealed 2 mm of enophthalmos on the left. After downward bending of the head the eye became exophthalmic by 12 mm and visual acuity decreased to finger counting. Neuro-ophthalmologic examination was otherwise normal. Magnetic resonance imaging revealed an intraconal hemangioma. Bluish, soft, rubber-like, compressible and slowly refilling hemangiomas on the inferior lip, sublingually, on the hand, and vagina led to the diagnosis of BRBNS. CONCLUSIONS: Intermittent visual loss may be the presenting symptom of an orbital hemangioma in BRBNS. Recognition of this rare condition is important because of its potentially life-threatening complications. PMID- 11438078 TI - Superficial temporal artery biopsy diagnostic for lung carcinoma. AB - PURPOSE: To report a unique finding of metastatic tumor cells in the lumen of a superficial temporal artery biopsy specimen in a patient with severe visual loss and optic neuropathy. METHODS: Case report. RESULTS: A 69-year-old man presented with anorexia, pancranial headaches, and acute unilateral visual loss. Ophthalmic examination showed a visual acuity of no light perception and an amaurotic pupil in the left eye. Fundus examination was normal. A superficial temporal artery biopsy, performed for presumed arteritic posterior ischemic optic neuropathy, demonstrated intraluminal cells with irregular nuclei and prominent nucleoli suggestive of metastatic malignancy. Further systemic evaluation uncovered a poorly differentiated adenocarcinoma of the lung with intravascular infiltration and multiple distant metastases. CONCLUSION: Extensive systemic intravascular tumor embolization may be associated with optic neuropathy. PMID- 11438091 TI - Free-radical mediated synthesis of enantiomerically pure, highly functionalized inositols from carbohydrates. AB - We report the synthesis, free-radical cyclization of precursors 1,2,7-trideoxy-7 iodo-3,4:5,6-di-O-isopropylidene-D-gluco-hept-1-enitol (1), methyl 7-O-acetyl-6-O benzyl-8-bromo-2,3,8-trideoxy-4,5-O-isopropylidene-D-gluco-oct-2-enonate (2) and 5-O-acetyl-4-O-benzyl-6-bromo-6-deoxy-2,3-O-isopropylidene-D-glucose-O benzyloxime (3), readily prepared from D-glucose, and some selected transformations of the carbocycles obtained from these intermediates. In compound 1 we have installed a terminal double bond and an iodide as radical acceptor and leaving group, respectively. Compounds 2 and 3 are epsilon-bromo aldehydes substituted with alpha,beta-unsaturated ester and oxime ether functions as radical traps, respectively. The tributyltin hydride mediated ring closure of these radical precursors have afforded a series of interesting, diverse and highly functionalized carbocycles which can be considered useful building blocks for the synthesis of branched-chain cyclitols, aminocyclitols and aminoconduritols. In these processes, a good chemical yield and high stereoselectivity has been found in the newly formed stereocenters. Particularly interesting has been the finding that the stereochemical outcome of the free radical cyclization is independent of the ratio of isomers (E or Z) in oxime ether 3. These results show the power and the state of art of this strategy for the stereocontrolled synthesis of enantiomerically pure inositols from carbohydrates. PMID- 11438092 TI - Synthesis and structure determination of some sugar amino acids related to alanine and 6-deoxymannojirimycin. AB - (5'R)-5'-Methyl-5'-[methyl (4S)-2,3-O-isopropylidene-beta-L-erythrofuranosid-4-C yl]-imidazolidin-2',4'-dione was synthesised starting from methyl 6-deoxy-2,3-O isopropylidene-alpha-D-lyxo-hexofuranosid-5-ulose applying the Bucherer-Bergs reaction. Its 5'-R configuration was confirmed by X-ray crystallography. Corresponding alpha-amino acid-methyl (5R)-5-amino-5-C-carboxy-5,6-dideoxy-alpha D-lyxo-hexofuranoside (alternative name: 2-[methyl (4S)-2,3-O-isopropylidene-beta L-erythrofuranosid-4-C-yl]-D-alanine) was obtained from the above hydantoin by acid hydrolysis of the isopropylidene group followed by basic hydrolysis of the hydantoin ring. Total deprotection afforded 5-C-carboxy-6-deoxymannojirimycin. Analogously, methyl (5S)-5-amino-5-C-carboxy-5,6-dideoxy-alpha-L-lyxo hexofuranoside and 5-C-carboxy-6-deoxy-L-mannojirimycin were prepared from the corresponding (5'S)-5'-methyl-5'-[methyl (4R)-2,3-O-isopropylidene-beta-D erythrofuranosid-4-C-yl]-imidazolidin-2',4'-dione starting from methyl 6-deoxy 2,3-O-isopropylidene-alpha-L-lyxo-hexofuranosid-5-ulose. PMID- 11438093 TI - Synthesis of propyl and 2-aminoethyl glycosides of alpha-D-galactosyl-(1-->3') beta-lactoside. AB - Propyl and 2-aminoethyl alpha-D-galactopyranosyl-(1-->3')-beta-lactosides (1 and 2) were prepared from the corresponding perbenzylated trisaccharide allyl glycoside 6 which, in turn, was obtained by methyl triflate promoted alpha galactosylation of benzylated allyl lactoside acceptor 4 with thiogalactoside 3. Transformation of the allyl moiety in compound 6 into 2-azidoethyl one was achieved by cleavage of the double bond followed by reduction into alcohol 9, subsequent mesylation, and mesylate-->azide substitution. Alternatively trisaccharide 2 was synthesized using alpha-galactosylation of selectively benzoylated 2-azidoethyl lactoside 19 with 3 as the key step. PMID- 11438094 TI - The hydrolytic and transferase action of alternanase on oligosaccharides. AB - Alternanase is an enzyme which endo-hydrolytically cleaves the alpha-(1-->3), alpha-(1-->6)-linked D-glucan, alternan. The main products are isomaltose, alpha D-Glcp-(1-->3)-alpha-D-Glcp-(1-->6)-D-Glc and the cyclic tetrasaccharide cyclo[- >6)-alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->3)-alpha-D-Glcp (1-->]. It is also capable of acting on oligosaccharide substrates. The cyclic tetrasaccharide is slowly hydrolyzed to isomaltose. Panose and the trisaccharide alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->3)-D-Glc both undergo transglycosylation reactions to give rise to the cyclic tetrasaccharide plus D-glucose, with panose being converted at a much faster rate. The tetrasaccharide alpha-D-Glcp-(1-->3) alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->4)-D-Glc is hydrolyzed to D-glucose plus the trisaccharide alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1-->6)-D-Glc. Alternanase does not act on isomaltotriose, theanderose (6(Glc)-O-alpha-D-Glcp sucrose), or alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->4)-alpha-D-Glc. The enzyme releases 4-nitrophenol from 4-nitrophenyl alpha-isomaltoside, but not from 4-nitrophenyl alpha-D-glucopyranoside, 4-nitrophenyl alpha-isomaltotrioside, or 4 nitrophenyl alpha-isomaltotetraoside. PMID- 11438095 TI - Rapid and simple preparation of N-linked oligosaccharides by cellulose-column chromatography. AB - As a means of preparing N-linked oligosaccharides from hydrazinolysates of glycoproteins in a rapid and simple manner, a method has been developed using cellulose-column chromatography. Hydrazinolysates of human IgG, containing a series of biantennary complex type oligosaccharides, were applied to a cellulose column equilibrated with (4:1:1, v/v) 1-butanol-ethanol-water. The N-linked oligosaccharides were eluted with (1:1, v/v) ethanol-water, and analyzed by HPLC in combination with sequential glycosidase digestion. The oligosaccharides, with or without sialic acid, were quantitatively recovered in the fraction eluted with (1:1, v/v) ethanol-water without UV-detectable contamination by impurities derived from protein or the cellulose. Other types of N-linked oligosaccharides of alpha1-acid glycoprotein (tetraantennary complex-type), ovalbumin (hybrid type), and ribonuclease B (high mannose-type) were also quantitatively prepared from the hydrazinolysates by elution of the cellulose column with (1:1, v/v) ethanol-water and these had as high a quality as those prepared by conventional paper chromatography. PMID- 11438096 TI - Characteristics of xyloglucan after attack by hydroxyl radicals. AB - It has been proposed that plant cell-wall polysaccharides are subject in vivo to non-enzymic scission mediated by hydroxyl radicals (-*OH). In the present study, xyloglucan was subjected in vitro to partial, non-enzymic scission by treatment with ascorbate plus H(2)O(2), which together generate -*OH. The partially degraded xyloglucan appeared to contain ester bonds within the backbone, as indicated by an irreversible decrease in viscosity upon alkaline hydrolysis. Aldehyde and/or ketone groups were also introduced into the polysaccharide by *OH-attack, as indicated by staining with aniline hydrogen-phthalate and by reaction with NaB(3)H(4). The introduction of ester and oxo groups supports the proposed sequence of reactions: (a) -*OH-mediated H-abstraction to produce a carbon-centred carbohydrate radical; (b) reaction of the latter with O(2); and (c) elimination of a hydroperoxyl radical (HO(2)*-). When the partially degraded xyloglucan was reduced with NaB(3)H(4) followed by acid hydrolysis, several 3H aldoses were detected ([3H]galactose, [3H]xylose, [3H]glucose, [3H]ribose and probably [3H]mannose), in addition to unidentified 3H-products (probably including anhydroaldoses). 3H-Alditols were undetectable, showing that few or no conventional reducing termini were introduced. Digestion of the NaB(3)H(4) reduced, partially degraded xyloglucan with Driselase released 25 times more [3H]Xyl-alpha-(1-->6)-Glc than Xyl-alpha-(1-->6)-[3H]Glc, suggesting that the xylose side-chains of the xyloglucan had been more heavily attacked by -*OH than the glucose residues of the backbone. The radioactive xyloglucan was readily digested by cellulase, yielding 3H-products in the hepta- to nonasaccharide range. A fingerprinting strategy for identifying -*OH-attacked xyloglucan in plant cell walls is proposed. PMID- 11438097 TI - Synthesis of D-xylopyranan by the ring-opening polymerization of 3-O-benzyl-alpha D-xylopyranose 1,2,4-orthopivalate. Attempts to synthesize a stereoregular polymer. AB - 3-O-Benzyl-alpha-D-xylopyranose 1,2,4-orthopivalate (1) was newly synthesized and polymerized under cationic polymerization reaction conditions in order to synthesize stereoregular (1-->4)-beta-D-xylopyranan. Although the polymerization of orthopivalate 1 was carried out under various reaction conditions, a non stereoregular polymer, but mainly consisting of (1-->4)-beta-xylopyranose units, was obtained. Comparing these results with those of glucose 1,2,4-orthopivalates, it was revealed that not only the substituents in the C-2 and C-3 positions, but also the CH(2)OR group in glucose 1,2,4-orthopivalate, largely contribute to (1- >4)-beta-glucosidic bond formation by the ring-opening polymerization. PMID- 11438098 TI - Molecular and crystal structures of N-aryl-beta-D-glycopyranosylamines from mannose and galactose. AB - The molecular and crystal structures of 12 N-aryl-beta-D-glycopyranosylamines have been determined by X-ray crystallography. Six of these are mannose derivatives, the N-p-bromophenyl (1), N-p-tolyl (2), N-m-chlorophenyl (3), N-p methoxyphenyl (4), N-o-chlorophenyl (5), and N-o-tolyl (6) derivatives that are formed by reaction with the corresponding substituted anilines. The remaining six are galactose derivatives, the N-phenyl (7), N-p-chlorophenyl (8), N-p bromophenyl (9), N-p-iodophenyl (10), N-p-nitrophenyl (11) and N-p-tolyl (12), derivatives prepared similarly. Compounds 1-3 assume the same packing arrangement. Compounds 4, 5, and 6 assume unique packing arrangements, although that assumed by 4 is closely related to that assumed by 1-3. Compounds 7-11 assume the same packing arrangement; that assumed by 12 is closely related to that assumed by 7-11. That the same packing arrangements can be maintained in spite of substantial changes in the electronic and steric nature of the substituent on the aryl ring reflects the strength of the hydrogen bond network connecting the monosaccharide portions of the molecules in the solid state. A hydrogen bonding motif found in all six mannose structures is a mutual interaction between translationally related molecules involving O-3-H...O-5 and O 6-H...O-4 hydrogen bonds. The recurrence of this motif throughout this group of mannosylamines suggests that it is an especially favorable interaction that might be expected to occur also in related macromolecular systems. PMID- 11438099 TI - From methyl mannosides to methyl octosides by a stepwise homologation with Grignard C(1) reagents. AB - A four-step procedure for homologation of methyl alpha-D-mannofuranoside and alpha-D-mannopyranoside was examined. The reactions consisted in (i) oxidation of the terminal hydroxymethyl group in a protected sugar derivative to an aldehyde; (ii) reaction with allyloxymethylmagnesium chloride (or (phenyldimethyl)silylmethyl-magnesium chloride); (iii) protection of the newly formed secondary alcohol group; (iv) deprotection of the terminal CH(2)OR (or oxidation of the CH(2)SiMe(2)Ph) group. From methyl alpha-D-mannosides, stereoisomeric DalphaD and LalphaD methyl heptosides and from them, methyl octosides of D-threo- and L-erythro-alpha-D-manno configuration were obtained. PMID- 11438100 TI - Structure of a galactomannan from the seeds of Cassia angustifolia Vahl. AB - Cassia angustifolia Vahl (family: Caesalpiniaceae) is a fast growing and spreading Indian shrub of which seeds, pods and leaves are extensively used for pharmaceutical applications. The seeds have been found to be an alternative source of commercial gums. The structural aspects of the galactomannans have been determined for a better understanding of its properties. The purified seed galactomannan contains mannose:galactose in a ratio of 2.90. The average molecular weight (M(w)) is 9.66x10(4) and the intrinsic viscosity (eta) is 209 mL/g. Methylation analysis, periodate oxidation, Smith degradation and 13C NMR studies confirm that the gum has the basic structure of legume galactomannans with a main chain of (1-->4)-linked beta-D-mannopyranosyl units to which single alpha-(1-->6)-D-linked galactopyranosyl units are attached through block pattern. PMID- 11438102 TI - Effects of Oxyrrhis marina (Dinoflagellata), bacteria and vitamin D(2) on population dynamics of Tisbe holothuriae (Copepoda). AB - Laboratory experiments were performed to determine which food combinations had the most favorable effect on population dynamics of the harpacticoid copepod Tisbe holothuriae. In addition to basic diets of TetraMin (T), rice (R) or a mixture of both (T+R), supplements consisting of the following were given: the heterotrophic dinoflagellate Oxyrrhis marina, a bacterial suspension and vitamin D(2). Results show a beneficial effect of bacteria at a given concentration on T. holothuriae, which is enhanced in combination with vitamin D(2). A higher concentration of bacteria together with vitamin D(2), however, has a reverse effect leading to greater mortality, lower sex ratio and number of ovigerous females, and a dramatic decrease in production of eggs, regardless of the basic diet. We conclude that different concentrations of a given bacterial community may have variable effects on the reproduction of harpacticoid copepods and that, in addition to the bacteria, vitamin D(2) may also play a role in invertebrate nutrition. PMID- 11438103 TI - The growth and cyst formation of a toxic dinoflagellate, Alexandrium tamarense, at low water temperatures in northeastern Japan. AB - A field survey was carried out in early spring to investigate the growth physiology and efficiency of cyst formation of Alexandrium tamarense in low water temperatures. A bloom of A. tamarense occurred in a stratified water column, formed by river inflow. The in situ growth rate estimated from daily cell abundance was high, 0.33 divisions day(-1), at 7.5-9 degrees C. New cysts began to be observed during the late growth phase. Maximum cyst flux (600 cysts cm(-2) day(-1)) was observed just after maximum cell abundance occurred. PO(4)-P Chl a( 1) gradually decreased and reached extremely low levels beyond the mid-growth phase of A. tamarense. As sinking cysts were also recognized at that time, it suggests cyst formation may have been induced by depletion of phosphorus source. The incidence of cyst formation (C.I) was 30%. The number of C.I was the same as reported previously for batch cultures under conditions suitable for vegetative growth. These results indicate that A. tamarense grows with considerably higher growth rate and transforms to cysts in high numbers, in low water temperatures in the field. PMID- 11438104 TI - Partitioning of food resources amongst 18 abundant benthic carnivorous fish species in marine waters on the lower west coast of Australia. AB - The volumetric contributions made by prey and plant material to the diets of 4 elasmobranch and 14 teleost species, collected seasonally by trawling from waters along ca. 200 km of the lower west coast of Australia, have been compared. These benthic carnivores, which were all abundant and collectively contributed 83% to the total number of fish caught, represented nine families (Urolophidae, Scorpaenidae, Triglidae, Platycephalidae, Sillaginidae, Carangidae, Gerreidae, Mullidae and Pempherididae). Some species were numerous in both shallow (5-15 m) and deeper (20-35 m) waters and in both northern and southern regions, whereas others were largely confined to one of these water depths or regions. Comparisons between the diets of the different species, which utilised data collected from individuals throughout the study area, demonstrated that the dietary composition of any given species was almost invariably significantly different from that of every other species. This partly reflected the fact that, while errant polychaetes, gammarid amphipods and tanaids were ingested by all species, their contributions to the diets of the different species varied. Furthermore, echinoderms contributed to the diets of just nine species, and this was substantial only in the case of two sillaginid species, while teleosts were never consumed by six species and only made a marked contribution to the diets of the single species of platycephalid. The diet of each species underwent size-related changes, reflecting a shift from the consumption by smaller fish of prey such as amphipods, mysids and copepods, to the ingestion by larger fish of prey such as polychaetes, carid decapods, isopods and small teleosts. The interspecific and intraspecific differences in dietary compositions would spread the food resources amongst and within species, thereby reducing the potential for competition for those resources within the fish community. Non-metric multi-dimensional scaling (MDS) ordination plots emphasised that the dietary compositions of species within each family possessed some obvious similarities, reflecting similarities in body and mouth morphology and feeding behaviour. However, the extent to which the dietary compositions of the different families were similar or different was often not related to the phylogenetic relationships amongst those families. Furthermore, while differences in mouth size and morphology could sometimes be used to account for differences amongst the diets of the full suite of species, this was not always the case. Thus, the ways in which species feed and use their ancillary feeding structures were also employed to help elucidate the basis for variations in diets. PMID- 11438105 TI - Growth, oxygen consumption, ammonia-N excretion, biochemical composition and energy content of Farfantepenaeus paulensis Perez-Farfante (Crustacea, Decapoda, Penaeidae) early postlarvae in different salinities. AB - Physiological responses of early postlarval shrimp Farfantepenaeus paulensis to different salinities were evaluated. Growth, net growth efficiency (K(2)), oxygen consumption, ammonia-N excretion, protein, lipid, carbohydrate, ash and energy content were determined in postlarval stages PL VI-VII and PL XIII-XV at 5 per thousand, 15 per thousand, 25 per thousand and 34 per thousand. PL VI-VII and PL XIII-XV correspond, in the wild, to the penetration of postlarvae into estuarine zones and the adoption of benthic habits, respectively. Postlarvae were gradually acclimated to the various salinity levels in the stage PL VI-VII and were maintained there until reaching the stage PL XIII-XV. Although development time was not affected by salinity, growth was decreased at 34 per thousand. Oxygen consumption was less affected by salinity while ammonia-N excretion presented a negative correlation with salinity. In both stages, protein content was lower at 34 per thousand. In PL VI-VII, lipid was decreased at 5 per thousand, while no significant difference in lipid content was observed in PL XIII-XV. Carbohydrate and ash levels did not vary in function of salinity levels. Energy content was reduced in PL XIII-XV at 34 per thousand. Net growth efficiency (K(2)) between PL VI-VII and PL XIII-XV presented higher values at 15 per thousand and 25 per thousand, and the lowest at 34 per thousand. Enhanced energy expenditure due to osmoregulation was not detected as denoted by oxygen consumption rates. Nitrogen metabolism seems to play a key role in maintaining body fluids concentration in relation to medium salinity. It is concluded that the high degree of euryhalinity in these stages of F. paulensis may be an adaptation to ontogenetic events such as migration to estuarine areas. For aquaculture purposes, it is recommended to maintain these postlarval stages at salinities between 15 per thousand and 25 per thousand rather than 34 per thousand. PMID- 11438106 TI - The effects of sediment-associated triorganotin compounds on the gills of the European flounder, Platichthys flesus (L.). AB - The effects of exposure to sediment-associated tri-n-butyltin chloride (TBT) and triphenyltin chloride (TPhT) were examined in the euryhaline European flounder, Platichthys flesus (L.). The effects were quantified by measuring the changes in sodium efflux, Na(+)/K(+)-ATPase activity and the numbers, areas and distribution of chloride cells in the gills of freshwater-adapted fish, following a rapid transfer to seawater. After transfer, the Na(+)/K(+)-ATPase activity and the sodium efflux significantly increased in both the TPhT and control groups but not in the TBT group. However, Na(+)/K(+)-ATPase activity and the sodium efflux in the TPhT group had returned to pre-salinity transfer levels by day 15 after the initial exposure to TPhT. Morphological changes in the numbers and areas of chloride cells, known to be associated with seawater adaptation, took place in the control group, i.e. there was a significant reduction in the number of lamellar chloride cells accompanied by an increase in the number of interlamellar chloride cells. There was a reduction in the numbers of lamellar chloride cells in the TBT-exposed group following transfer to seawater but the mean number was significantly higher than the control group by the end of the experiment. In the TPhT-exposed group, the reduction was not significantly different to that seen in the control group. By the end of the experiment, both organotin-exposed groups had significantly lower mean numbers of interlamellar chloride cells than the control group. Before transfer to seawater, the mean areas of lamellar and interlamellar chloride cells of all three groups were not significantly different. On transfer, the mean areas of lamellar chloride cells in the control group became significantly smaller than the mean areas of the organotin groups. There was no significant difference in the mean areas of interlamellar chloride cells in the control and TBT groups between the start and finish of the experiment but there was a significant increase in the mean area of TPhT-treated animals at the end of the experiment when compared to the control group. The results presented in this study lead to the conclusion that tri-n-butyltin chloride and triphenyltin chloride in sediments are capable of significantly disrupting both the physiological as well as morphological components of ionic regulatory functions of an estuarine fish, at concentrations currently found in estuarine sediments. PMID- 11438107 TI - Schooling affects the feeding success of Australian salmon (Arripis trutta) when preying on mysid swarms (Paramesopodopsis rufa). AB - When feeding on mysid swarms (Paramesopodopsis rufa), juvenile Australian salmon (Arripis trutta) had higher rates of successful attacks when foraging in a group of six fish (55% total advances) than when foraging alone (39% total advances). Six schooling fish had lower approach rates than solitary fish (25% and 37% of total advances, respectively). This result indicated that schooling fish were better at reducing the confusion effect of swarming prey, resulting in more efficient feeding. In larger areas, schools achieved higher rates of successful attacks (19 prey/fish in the large tank, compared with 11 prey/fish in the smaller tank). There was no influence on the feeding success of individual fish when changes were made to the number of prey presented to each fish. Nearest neighbour distances were smallest in the absence of prey, and increased with the introduction of prey and again in an attack sequence. Six fish schooled more cohesively than three fish, indicating increased benefits of schooling in larger groups that contribute to advanced vigilance and foraging techniques. PMID- 11438108 TI - The photosynthetic capacity of the seagrass Posidonia oceanica: influence of nitrogen and light. AB - Photosynthetic capacity of the seagrass Posidonia oceanica was determined in two contrasting nutrient zones along a bathymetric gradient. Photosynthetic capacity was two times higher in the relatively eutrophic zone compared to the highly oligotrophic zone. The same trend was observed in the chlorophyll a+b and tissue nitrogen content. Photosynthetic capacity and chlorophyll a+b decreased with irradiance in the oligotrophic zone while increased in the eutrophic zone. Tissue nitrogen content followed the same trend in the oligotrophic zone and did not present any trend in the eutrophic zone. Overall, this study has shown the importance of large scale sampling when assessing metabolic parameters such as photosynthetic capacity where large differences have been observed associated to the nutrient regime. According to our results, in oligotrophic zones nitrogen seems to govern photosynthetic capacity and there is a coherent pattern between nitrogen content, pigments and photosynthetic capacities. In eutrophic zones, light is the dominant regulator and there is a coherent pattern between light and photosynthetic capacities. PMID- 11438109 TI - Isolation and characterization of the metamorphic inducer of the common mud crab, Panopeus herbstii. AB - Several items from the natural habitat of adult Panopeus herbstii were examined to determine if they had the ability to produce a metamorphic cue. These included adult conspecifics, natural rock/shell substratum, the co-occurring species Hemigrapsus sanguineus and bacterial biofilms. Adult conspecifics, H. sangineus and natural rock/shell all accelerated metamorphosis. However, adult conspecifics accelerated metamorphosis to the greatest extent. The cue associated with adult conspecifics was found to be water-soluble, stable following boiling and freezing, and of relatively small molecular size (<1 kDa). Furthermore, the cue appears to be produced from the conspecifics themselves, rather than from biofilms colonizing the surfaces of the crabs. The results of this experiment suggest that postlarvae of P. herbstii are able to distinguish suitable habitat through chemical signals, thus greatly increasing their chances for survival. PMID- 11438110 TI - Depressed mood, physician-rated prognosis, and comorbidity as independent predictors of 1-year mortality in consecutive medical inpatients. AB - OBJECTIVE: To determine the independent effects of depressed mood and markers of medical disease severity on mortality in consecutive medical inpatients. METHODS: Consecutive general medical inpatients were asked to complete the Hospital Anxiety and Depression Scale (HADS) at admission. Prognostic indicators were obtained from patients' records and physicians' ratings. The study endpoint was mortality from all causes at 1 year. RESULTS: The baseline assessment was completed by 575 patients (87.7%). Survival data were available for 572 of these (86 deaths). HADS depression scores and several physical risk indicators predicted mortality. In multivariate analyses, physicians' rating of prognosis was the best predictor of mortality [adjusted odds ratio (OR) 3.6; 95% confidence interval (CI), 2.5--5.4]. Other independent predictors included a principal diagnosis of hemato-oncological disease, comorbidity scores, and HADS depression (adjusted OR 1.75; 95% CI, 1.10--2.79). CONCLUSION: Our data demonstrate an independent prognostic effect of depressed mood on mortality in general medical inpatients. Screening for depression may improve risk stratification in these patients over and above that obtained by routinely available physical parameters and physicians' clinical judgement. PMID- 11438111 TI - Psychological characteristics of women presenting with breast pain. AB - OBJECTIVE: Extensive evidence links unexplained physical symptoms in adults with childhood abuse. This study investigated a possible link between unexplained breast pain, recalled childhood abuse and emotional and somatic distress. METHODS: Consecutive female out-patients presenting with breast pain were categorised as treatment-resistant (n=20), newly diagnosed and requesting treatment (n=37), or newly diagnosed but reassured and not seeking treatment (n=32) and compared with pain-free patients with breast lumps (n=31). RESULTS: All breast pain groups were more anxious and depressed, somatised more and recalled a higher incidence of emotional abuse by comparison with breast lump patients. Logistic regression confirmed that emotional abuse and anxiety were independently associated with breast pain. CONCLUSION: Clinical management of patients who present breast pain should be sensitive to the evidence that pain is a marker of emotional abuse in some women and is associated with widespread somatic and emotional distress. PMID- 11438112 TI - Interpersonal behavior and resting blood pressure in college women: a daily monitoring study. AB - Psychological factors have long been implicated in the development of hypertension. Most studies exploring this relationship employed questionnaires administered on only one occasion. OBJECTIVE: to evaluate the relation between blood pressure and social behavior in the natural environment over an extended period of time. METHOD: 40 healthy young adult women at varying risk for hypertension were asked to record their behavior and affect following social interactions, three times a day for 32 days. Behaviors were representative of the interpersonal circumplex: dominance, submissiveness, agreeableness, and quarrelsomeness. RESULTS: Casual blood pressure was significantly correlated with submissive behavior (r=.45, P<.05) and inversely correlated with agreeable behavior (r=-.35, P<.05). ANOVAs confirmed these findings and further revealed that offspring of hypertensives with relatively elevated blood pressure were most quarrelsome. CONCLUSION: These results concur with the literature on the relationship between hostility and blood pressure but also suggest the importance of submissiveness in college women. PMID- 11438113 TI - Psychological distress and disability in patients with vertigo. AB - OBJECTIVE: Vertigo is an extremely debilitating experience for the patient, especially during attacks; it is neither easy to identify nor control. The importance of psychosomatic factors has already been widely studied and discussed. In particular, it has been shown that stress factors are relevant in setting off episodes of dizziness, but there is no agreement if the presence of distress might influence the vestibular disability. METHODS: This study is concerned with evaluating the quality of life (QOL) in a group of 206 patients suffering from vertigo and 86 control patients, using the UCLA-Dizziness Questionnaire (UCLA-DQ) scale. The results were correlated with those achieved using the Hospital Anxiety and Depression Scale (HADS) psychometric test. RESULTS: What is clear is that, in patients suffering from vertigo as regards those who are not, there is a significant amount of anxiety and depression distress, especially in female subjects. There appears to be no relationship between psychological change and the various forms of clinical vertigo. In terms of the QOL parameter, what emerges is that, from a statistical point of view, fear of becoming dizzy is most closely correlated with the perception of disability. CONCLUSIONS: There is a also a need for psycho-education here in collaboration with the E.N.T. specialist so that the patient can learn to recognise his/her medical condition and be aware of the factors that primarily contribute to the deterioration of their QOL. PMID- 11438114 TI - Postoperative fatigue is a component of the emotional response to surgery: results of multivariate analysis. AB - OBJECTIVE: To test the theory that postoperative fatigue is an aspect of the emotional, not physiological, response to surgery, we examined whether fatigue is a component of subjective experience after surgery and whether it is related to subjective physical or emotional state. METHODS: Patients (N=160) undergoing hip or knee arthroplasty were assessed preoperatively, 1 and 7 days and 1 and 6 months postoperatively using multiple indicators of fatigue and subjective emotional and physical state. Covariance structure modeling was used to find out the structure of patients' experience on each occasion. RESULTS: At each time, data indicated four latent variables: negative mood, positive mood, dysfunction and pain. Scales measuring fatigue indicated negative or positive mood but were unrelated to dysfunction and pain. DISCUSSION: In surgical patients, the language of fatigue and energy describes empirically distinct components of emotional state. Explanations for postoperative fatigue should therefore be sought in emotional, not physiological, mechanisms. PMID- 11438115 TI - The Skin Picking Scale: scale construction and psychometric analyses. AB - OBJECTIVE: This paper reports on the development of the Skin Picking Scale (SPS), a six-item paper-and-pencil measure for the assessment of skin picking. METHODS: 28 severe self-injurious and 77 non-self-injurious skin pickers initially completed an eight-item severity scale modeled after the Yale--Brown Obsessive- Compulsive Scale (Y-BOCS). RESULTS: Group comparisons and part--whole correlations for individual scale items resulted in a six-item scale with a total score range of 0--24. A Cronbach's alpha coefficient of.80 indicated moderate internal consistency for the scale. Construct validity was demonstrated by significant correlations between SPS total scale scores and self-reported average duration of skin picking episodes. Significant correlations were also reported between SPS total scale scores and both Beck Depression Inventory (BDI) and Beck Anxiety Inventory (BAI) scores, as well as self-reported intensity of emotions during the picking process. Lastly, sensitivity and specificity analyses suggest that a SPS cut-off score of 7 differentiates severe self-injurious and non-self injurious skin pickers. CONCLUSION: The SPS is a valid and reliable self-report scale for the assessment of severity in medical and psychiatric patients who endorse skin picking. PMID- 11438116 TI - Mutant and genetically modified mice as models for studying the relationship between aging and carcinogenesis. AB - Increased interest is emerging in using mouse models to assess the genetics of aging and age-related diseases, including cancer. However, only limited information is available regarding the relationship between aging and spontaneous tumor development in genetically modified mice. Analysis of various transgenic and knockout rodent models with either a shortened or an extended life span, provides a unique opportunity to evaluate interactions of genes involved in the aging process and carcinogenesis. There are only a few models which show life span extension. Ames dwarf mutant mice, p66(-/-) knockout mice, alpha MUPA and MGMT transgenic mice live longer than wild-type strains. The incidence of spontaneous tumors in these mutant mice was usually similar to those in controls, whereas the latent period of tumor development was increased. Practically all models of accelerated aging showed increased incidence and shorter latency of tumors. This phenomenon has been observed in animals which display a phenotype that more closely resembles natural aging, and in animals which manifest only some features of the normal aging process. These observations are in agreement with an earlier established positive correlation between tumor incidence and the rate of tumor incidence increase associated with aging and the aging rate in a population. Thus, genetically modified animals are a valuable tool in unravelling mechanisms underlying aging and cancer. Systemic evaluation of newly generated models should include onco-gerontological studies. PMID- 11438117 TI - Microanatomical changes of intracerebral arteries in spontaneously hypertensive rats: a model of cerebrovascular disease of the elderly. AB - Changes occurring in intracerebral arteries of 24-week-old spontaneously hypertensive rats (SHR) compared with age-matched normotensive Wistar-Kyoto (WKY) rats were assessed using microanatomical techniques associated with image analysis. Morphometric parameters investigated included arterial diameter, lumen area, wall area, and wall-to-lumen ratio. Intracerebral arteries (lumen diameter>46 microm) and arterioles (lumen diameter 46-10 microm) of frontal cortex, striatum, and hippocampus were examined. In frontal cortex of SHR arterial wall hypertrophy and luminal narrowing were observed. In striatum, an increase of wall area not accompanied by luminal narrowing predominates resulting in arterial hypertrophy without vasoconstriction. In hippocampal arteries of SHR, luminal narrowing, without changes of wall area was found indicating the occurrence of remodeling. In brain areas investigated, hypertensive changes affected primarily arterioles. The demonstration of a sensitivity of intracerebral arteries to hypertension suggests that changes of these vessels may represent a cause of brain structural alterations occurring in hypertension. The specificity of alterations occurring in intracerebral arteries of brain areas investigated may account for the different localization of cerebral lesions in cerebrovascular disease. The possibility that microanatomical changes developed in intracerebral arteries of SHR may represent a model of cerebrovascular disease of the elderly is discussed. PMID- 11438118 TI - Cytokine production by stimulated mononuclear cells did not change with aging in apparently healthy, well-nourished women. AB - Aging is often associated with a dysregulation of the immune system. We examined mitogen-stimulated production of interleukin (IL)-2 and proinflammatory cytokines, IL-1beta and IL-6, in apparently healthy and generally well-nourished old versus young women. Subjects were screened for health using the SENIEUR protocol and a panel of laboratory tests for inflammation, as well as for the adequacy of nutritional status using criteria related to undernutrition, and protein, iron, vitamin B(12), and folate status. Young (n=26, age: 20-40 years) and old (n=44, age: 62-88 years) cohorts did not differ on the number of circulating monocytes, granulocytes, B (CD19+) cells, and T (CD3+, CD4+, and CD8+) cells. No differences (P>0.10) were seen between the two age groups in IL 2, IL-1beta and IL-6 levels in whole blood cultures at 48 h after stimulation with PHA (5 mg/l). Furthermore, no age-related differences were noted in the absolute amounts (pg) of IL-1beta and IL-6 after normalizing for circulating monocytes, B cells, or T cells (P>0.10). Similarly, no age-related decline in absolute amount of IL-2 (pg) after normalizing for circulating T cells was noted (P>0.10). Thus, contrary to most previous reports, our results do not support an increase in the production of proinflammatory cytokines IL-1beta and IL-6, and a reduced production of IL-2 with aging when health and nutritional status are maintained. These findings support our previous results of no change in monocyte function and few alterations in acquired immune response in a carefully selected group of healthy and well-nourished elderly women. PMID- 11438119 TI - Sequential changes in MR water proton relaxation time detect the process of rat brain myelination during maturation. AB - For better understanding of the behavior of water molecules in the animal brain, changes in magnetic resonance water proton relaxation processes were studied in the rat during maturation. Midbrains of male Wistar rats were removed at various time points ranging from 2 to 70 days after birth. Changes in relaxation time (water proton longitudinal relaxation time by the inversion recovery, and water proton transverse relaxation time by the spin echo and the Carr-Purcell-Meiboom Gill pulse sequence (CPMG)) and water content were then determined for various stages of brain development. During maturation both water proton longitudinal relaxation time and water proton transverse relaxation time values decreased and this finding paralleled the decline in water content. Using the CPMG pulse sequence, the transverse relaxation time values were observed to separate into two components after 21 days. Morphologically, the most prominent change at the matured stage of midbrain development in the rat is myelination. Water proton relaxation time, which can be estimated using the CPMG pulse sequence, showed a close correlation with myelination in the central nervous system. PMID- 11438120 TI - Acute neutral endopeptidase inhibition is natriuretic in old rats. AB - Neutral endopeptidase 24.11 (NEP) inhibitors prevent breakdown of atrial natriuretic peptide (ANP), and may be useful therapeutically, in sodium overload states as often occurs in the aged. However, age-dependent changes in ANP/NEP may limit the activity of these agents in the elderly. To investigate this we conducted experiments in young, middle aged and old conscious male rats, studied in the baseline euvolemic state and during acute NEP inhibition (NEPI). NEPI produced a marked increase in sodium excretion (>100%) in all groups, regardless of age. A selective, potassium sparing effect was also seen, only in the middle aged and old rats. Although baseline hemodynamics were affected by age with mean blood pressure, BP, and renal vascular resistance (RVR) being higher in old versus young (131+/-5 vs. 115+/-3 mmHg; P<0.05 and 29+/-3 vs. 20+/-1 mmHg/ml per min per 100 g body weight (BW); P<0.02, respectively); NEPI produced similar mild pressor and significant renal vasoconstrictor effects in all age groups. Despite the tendency of NEPI to reduce renal perfusion, this is an effective method of increasing sodium excretion in all age groups while the potassium sparing actions seen selectively in the older rats may increase the usefulness of NEPI as a diuretic agent for the elderly. PMID- 11438121 TI - Long-term insulin-like growth factor-I expression in skeletal muscles attenuates the enhanced in vitro proliferation ability of the resident satellite cells in transgenic mice. AB - Insulin-like growth factor-I (IGF-I) overexpression for 1-month in mouse skeletal muscle increases satellite cell proliferation potential. However, it is unknown whether this beneficial enhancement by IGF-I expression would persist over a longer-term duration in aged mice. This is an important issue to address if a prolonged course of IGF-I is to be used clinically in muscle-wasting conditions where satellite cells may become limiting. Using the IGF-I transgenic (IGF-I Tg) mouse that selectively expresses the IGF-I transgene in striated muscles, we found that 18-months of continuous IGF-I overexpression led to a loss in the enhanced in vitro proliferative capacity of satellite cells from Tg skeletal muscles. Also 18-month-old IGF-I Tg satellite cells lost the enhanced BrdU incorporation, greater pRb and Akt phosphorylations, and decreased p27(Kip1) levels initially observed in cells from 1-month-old IGF-I Tg mice. The levels of those biochemical markers reverted to similar values seen in the 18-months WT littermates. These findings, therefore, suggest that there is no further beneficial effect on enhancing satellite cell proliferation ability with persistent long-term expression of IGF-I in skeletal muscles of these transgenic mice. PMID- 11438124 TI - Childhood obesity: an emerging public-health problem. PMID- 11438122 TI - Mutation frequency and type during ageing in mouse seminiferous tubules. AB - Mutations arise in the germline by errors of replication, recombination and repair, and the movement of transposable elements. Transgenic mice bearing reporter genes such as lacZ have proven useful for measurements of spontaneous and induced mutation frequencies, as well as studies of the effects of ageing. In this study, testicular DNA from lacZ transgenic mice was examined for age-related effects on mutation frequency and type. The recovered transgene was tested for simple substitutions and rearrangements including transposition of endogenous mobile elements. There was no evidence for either an age-related accumulation of mutations, or for the insertion of retrotransposons into the lacZ reporter gene in the testis. We conclude that the frequency of retrotransposition of several mouse mobile elements into the lacZ reporter gene is less than 3.73x10(-8). This is significantly less than the known frequency of approximately 7% of all spontaneous mutations in the mouse being due to retrotransposition of these elements. PMID- 11438125 TI - Birthweights, maternal cardiovascular events, and Barker hypothesis. PMID- 11438126 TI - Leukotriene antagonists and symptom control in chronic persistent asthma. PMID- 11438127 TI - Extracellular matrix and tenascin-C in pathogenesis of breast cancer. PMID- 11438128 TI - Time to take soundings in acute rheumatic fever. PMID- 11438129 TI - Is vitamin K deficiency a risk factor for osteoporosis in Crohn's disease? PMID- 11438131 TI - Pregnancy complications and maternal risk of ischaemic heart disease: a retrospective cohort study of 129,290 births. AB - BACKGROUND: Individuals who are small at birth are at increased risk of ischaemic heart disease (IHD) in later life. One hypothesis to explain this association is fetal adaptation to a suboptimum intrauterine environment. We investigated whether pregnancy complications associated with low birthweight are related to risk of subsequent IHD in the mother. METHODS: Routine discharge data were used to identify all singleton first births in Scotland between 1981 and 1985. Linkage to the mothers' subsequent admissions and deaths provided 15--19 years of follow up. The mothers' risks of death from any cause or from IHD and admission for or death from IHD were related to adverse obstetric outcomes in the first pregnancy. Hazard ratios were adjusted for socioeconomic deprivation, maternal height and age, and essential hypertension. FINDINGS: Complete data were available on 129,920 (95.6%) eligible deliveries. Maternal risk of IHD admission or death was associated with delivering a baby in the lowest birthweight quintile for gestational age (adjusted hazard ratio 1.9 [95% CI 1.5--2.4]), preterm delivery (1.8 [1.3--2.5]), and pre-eclampsia (2.0 [1.5--2.5]). The associations were additive; women with all three characteristics had a risk of IHD admission or death seven times (95% CI 3.3--14.5) greater than the reference category. INTERPRETATION: Complications of pregnancy linked to low birthweight are associated with an increased risk of subsequent IHD in the mother. Common genetic risk factors might explain the link between birthweight and risk of IHD in both the individual and the mother. PMID- 11438130 TI - Birthweight, early environment, and genetics: a study of twins discordant for acute myocardial infarction. AB - BACKGROUND: Epidemiological studies that used birthweight as a crude marker of fetal growth have suggested that low birthweight is associated with increased risk of coronary heart disease. Through investigation of this association within same-sexed twin pairs, confounding by genetic and early environmental factors can be greatly decreased. We undertook a case-control study in twins discordant for acute myocardial infarction (AMI). METHODS: The case-control study was nested within the population-based Swedish Twin Registry and linked with the national cause-of-death and hospital-discharge registries. We manually retrieved birth records containing information on birth and maternal characteristics for 132 same sexed twin pairs discordant for AMI and 118 individually matched control twin pairs. FINDINGS: In comparisons between AMI cases and external matched control twins, cases had significantly lower birthweight (mean 2556 [SD 500] vs 2699 [530] g, p=0.04), birth length (47.1 [2.8] vs 47.9 [2.7] cm, p=0.04), and head circumference (33.0 [1.8] vs 33.5 [2.0] cm, p=0.03) than controls. In within-pair comparisons between AMI cases and healthy co-twins, no significant differences in birth measurements were found (birthweight 2458 [510] vs 2534 [530] g, p=0.73; birth length 47.1 [2.8] vs 47.2 [2.8] cm, p=0.91; head circumference 33.0 [1.7] vs 33.0 [1.8] cm, p=0.92). INTERPRETATION: The lack of an association between birth characteristics and AMI within twin pairs suggests that previously reported associations may be influenced by genetic and early environmental factors, or possibly, by unmeasured maternal factors that operate independently of birthweight. PMID- 11438132 TI - Addition of leukotriene antagonists to therapy in chronic persistent asthma: a randomised double-blind placebo-controlled trial. AB - BACKGROUND: Controlled trials suggest that leukotriene receptor antagonists can improve lung function and reduce requirement for oral or inhaled corticosteroids in patients with asthma. We aimed to assess whether montelukast, a leukotriene receptor antagonist, can improve symptoms or lung function in patients with chronic asthma with symptoms already taking corticosteroids. METHODS: We did a double-blind, placebo-controlled, crossover, randomised add-on study in which 100 patients with asthma and symptoms despite treatment with inhaled corticosteroids and additional therapy were given 10 mg montelukast sodium for 14 days in an outpatient clinic setting. Outcome measures were symptoms and peak flow diaries. FINDINGS: 72 patients had diary data for analysis. Compared with placebo, addition of montelukast did not result in any significant change in symptom scores (mean difference between the last 7 days of each treatment period 0.05; 95% CI --0.86 to 1.14), rescue inhaled beta(2) agonist use (mean difference in puffs per day 0.41; -0.29 to 0.57), or twice daily peak expiratory flow (PEF) measurements (mean difference in morning PEF 1.18 L/min; -14.29 to 17.14), and mean difference in evening peak flow (-0.50; -17.42 to 12.86). When treatment response was defined as a 15% or greater increase in mean peak flow readings, there were four responders to montelukast and seven responders to placebo. INTERPRETATION: Based on PEF data from our previous studies of a similar patient group we would have expected to detect changes of more than 5%. Used as additional therapy in a hospital outpatient clinic setting, montelukast did not provide such additional benefit in patients with moderate or severe asthma. PMID- 11438133 TI - Clinical picture: fuel on the fire. PMID- 11438134 TI - Narrow-band ultraviolet B and broad-band ultraviolet A phototherapy in adult atopic eczema: a randomised controlled trial. AB - BACKGROUND: Narrow-band ultraviolet B (UVB) is an effective treatment for psoriasis, and open studies suggest that this phototherapy might improve atopic eczema. We did a randomised controlled trial to compare narrow-band UVB, UVA, and visible light phototherapy as second-line, adjunctive treatments in adult patients with moderate to severe atopic eczema. METHODS: Phototherapy was administered twice a week for 12 weeks. 26 patients were randomly assigned narrow band UVB, 24 were assigned UVA, and 23 visible fluorescent light. The primary endpoints were change in total disease activity (sum of scores at six body sites) and change in extent of disease after 24 treatments compared with baseline. Data were analysed by the method of summary measures. FINDINGS: 13 patients withdrew or were excluded from analysis. Mean reductions in total disease activity over 24 treatments in patients who received narrow-band UVB and UVA, respectively, were 9.4 points (95% CI 3.6 to 15.2) and 4.4 points (-1.0 to 9.8) more than in patients who received visible light. Mean reductions in extent of disease after 24 treatments with narrow-band UVB and UVA were 6.7% (1.5 to 11.9) and -1.0% ( 5.3 to 3.3) compared with visible light. A small proportion of patients developed erythema after phototherapy or had a flare in their eczema sufficient to withdraw from treatment. INTERPRETATION: Narrow-band UVB is an effective adjunctive treatment for moderate to severe atopic eczema, and the treatment is well tolerated by most patients. PMID- 11438135 TI - Enhanced contact tracing and spatial tracking of Mycobacterium tuberculosis infection by enumeration of antigen-specific T cells. AB - BACKGROUND: Identification of individuals latently infected with Mycobacterium tuberculosis is an important part of tuberculosis control. The current method, the tuberculin skin test (TST), has poor specificity because of the antigenic cross-reactivity of purified protein derivative (PPD) with M bovis BCG vaccine and environmental mycobacteria. ESAT-6 is a secreted antigen that is highly specific for M tuberculosis complex, but is absent from M bovis BCG. With an enzyme-linked immunospot (ELISPOT) assay for interferon gamma, we have identified ESAT-6-specific T cells as an accurate marker of M tuberculosis infection. METHODS: We did a prospective, masked study of 50 healthy contacts, with varying but well defined degrees of exposure to M tuberculosis, who attended an urban contact-tracing clinic. We assessed and compared the efficacy of our assay and TST for detection of symptomless infected individuals by correlation of test results with the degree of exposure to an infectious index case. FINDINGS: The ESAT-6 ELISPOT assay results had a strong positive relation with increasing intensity of exposure (odds ratio=9.0 per unit increase in level of exposure [95% CI 2.6--31.6], p=0.001), whereas TST results had a weaker relation with exposure (1.9 [1.0--3.5], p=0.05). By contrast, ELISPOT results were not correlated with BCG vaccination status (p=0.7), whereas TST results were significantly more likely to be positive in BCG-vaccinated contacts (12.1 [1.3--115.7], p=0.03). INTERPRETATION: This new antigen-specific T cell-based assay could allow more accurate identification of symptom-free individuals recently exposed to M tuberculosis, and thereby help to improve tuberculosis control. PMID- 11438136 TI - A language barrier, abdominal pain, and double vision. PMID- 11438137 TI - Platelet derived CD154 (CD40 ligand) and febrile responses to transfusion. PMID- 11438138 TI - Association between sciatica and Propionibacterium acnes. PMID- 11438139 TI - Plasminogen binds to disease-associated prion protein of multiple species. PMID- 11438140 TI - Developing countries tackle reproductive health. PMID- 11438147 TI - Breaking up the bacterial happy home. PMID- 11438148 TI - Patients' Bill of Rights moves ahead on US political agenda. PMID- 11438151 TI - Kenya allows parallel importation of drugs. PMID- 11438153 TI - Abortion boat faces legal complications. PMID- 11438154 TI - Pakistan announces national plan to tackle key health issues. PMID- 11438157 TI - Immunogenetics and genomics. AB - Immunogenetic analysis of disease susceptibility has been encouraged by the identification of strong HLA associations with several diseases of uncertain cause. Weaker HLA associations exist with a large number of infectious and non infectious diseases and the mechanisms of these effects are beginning to be uncovered. Extensive analyses of non-HLA immunogenetic variants have also been undertaken and associations with a variety of genes identified. Genetic linkage analysis of multicase families has recently identified new major susceptibility loci for a few immunologically determined common diseases. However, the greatest potential for the future lies in genome-wide searches for susceptibility genes that individually might have quite modest effects but cumulatively have a large impact on individual risk. This new era of immunogenomics promises to provide key insights into disease pathogenesis and identify multiple molecular targets for intervention strategies. PMID- 11438158 TI - Raynaud's phenomenon. AB - Raynaud's phenomenon is characterised by episodic vasospasm of the fingers and toes typically precipitated by exposure to cold. Mild Raynaud's is common and is not usually a harbinger of clinically important disability; its onset, however, can be startling and uncomfortable for patients, and the well recognised association in some cases with systemic rheumatic conditions often precipitates aggressive assessments for underlying diseases. Advances in vascular physiology have shed light on the role of the endothelium as well as endothelium-independent mechanisms in the altered vasoregulation of Raynaud's. We review clinical aspects of the disorder and new insights with respect to pathophysiology, and we discuss potential new therapeutics based on the disease mechanism, such as prostacyclin analogues, serotonin antagonists, and calcitonin gene-related peptides. PMID- 11438159 TI - Francois de la Peyronie and the disease named after him. PMID- 11438167 TI - Enhanced susceptibility to Theiler's virus-induced demyelinating disease in perforin-deficient mice. AB - Theiler's virus induces immune-mediated demyelinating disease similar to human MS in susceptible mice. Though the MHC class II-restricted T cell response is critical, susceptibility/resistance is also associated with a MHC class I haplotype. Here we report that perforin-deficient C57BL/6 mice (pKO) are susceptible to demyelination and develop clinical disease. The levels of primary demyelination, proliferation, Th1 responses, and viral load were also markedly enhanced. In addition, immunization of pKO mice with UV-inactivated virus further enhanced clinical incidence and accelerated the disease course. Thus, perforin is most likely involved in viral clearance, hence protection from the disease. PMID- 11438168 TI - Local immune regulation in the central nervous system by substance P vs. glutamate. AB - The response of parenchymal microglia to interferon-gamma (IFN-gamma) varies across the brain. To ask if local neurochemicals contribute to site-specific control, the influence of substance P (SP) and glutamate was evaluated in brainstem vs. hippocampus. In brainstem, stereotaxic injection of SP increased class II MHC upregulation by IFN-gamma, while a SP receptor antagonist (Spantide I) prevented it. In hippocampus, where the baseline response to IFN-gamma was lower, SP was ineffective, but blocking glutamate enhanced the response in a proportion of rats. Attempts to understand and control immune activity in the CNS should take the local neurochemical environment into account. PMID- 11438169 TI - Role of hypothalamic-pituitary axis in morphine-induced alteration in thymic cell distribution using mu-opioid receptor knockout mice. AB - Mu-opioid receptor knockout mice (MORKO), were used to address two questions: (1) if morphine induced decrease in thymic weight and cell distribution is mediated by the mu-opioid receptor and (2) the role of corticosteroids in morphine mediated alteration in thymic cell distribution. Our result show that morphine mediated increase in plasma corticosterone is mediated by the mu-opioid receptor since morphine at doses as high as 25 mg/kg-body weight does not increase plasma corticosterone levels in the MORKO. In addition, we have also shown that morphine treatment results in the differentiation of CD4+CD8+ (double positive cells) to single positive CD4+ cells while dexamethasone treatment results in the deletion of CD4+CD8+ (double positive) cells. PMID- 11438170 TI - Changes with aging in the modulation by neuropeptide Y of murine peritoneal macrophage functions. AB - Some age-related changes in immune function may be due, at least in part, to a disturbance in the communication between the nervous and immune systems. In the present work, the effects in vitro of neuropeptide Y (NPY) (10(-13) to 10(-7) M) on different peritoneal macrophage functions (adherence to substrate, chemotaxis, phagocytosis, superoxide anion production, and the release of TNFalpha and IL 1beta) have been studied on cells from young (12+/-2 weeks), adult (24+/-2 weeks), mature (50+/-2 weeks) and old (72+/-2 weeks) BALB/c mice. The specificity of these actions was confirmed using two C-terminal fragments of NPY, and the intracellular messengers (protein kinase C and cAMP) involved in the action of the neuropeptide were also analyzed. The results show that the functions studied change with aging and that the effects of NPY on each function, which are carried out through specific receptors, as well as on intracellular pathway, differ depending on age, maintaining the immune functions at physiologically adequate levels in old animals. PMID- 11438171 TI - Curative effects of recombinant human Interleukin-6 in DA rats with protracted relapsing experimental allergic encephalomyelitis. AB - We have studied the effects of treatment with recombinant human (rh)IL-6 on clinical, histological and immunological parameters of protracted relapsing (PR) experimental allergic encephalomyelitis (EAE) in DA rats. rhIL-6 (50 microg/rat subcutaneously/day) was given under three different regimens, as early prophylaxis, from 1 day prior to 14 days after immunization, in late prophylaxis, from day +7 until day 21 post-immunization (p.i.) and therapeutically to rats with clinical signs of EAE from day 14 to day 28 p.i. Although rhIL-6 failed to modulate the course of PR-EAE when administered as the early prophylactic regimen, it exerted clear-cut favourable effects on the course of the disease if was administered either as later prophylactic or as therapeutic treatment. Under these conditions, rhIL-6 accelerated recovery from EAE attacks and reduced/milded subsequent EAE episodes as compared to either PBS- or heat-inactivated rhIL-6 treated control rats. In agreement with this clinical effect, relative to PBS treated rats, the animals injected with rhIL-6 exhibited lower numbers of MHC class II(+) and CD4(+) cells in their spinal cords. rhIL-6-treatment also profoundly modulated the endogenous cytokine network, the treated rats displaying increased numbers of spleen cells expressing mRNA transcripts of the anti inflammatory cytokines IL-10 and TGF-beta along with simultaneously reduced numbers of mRNAs for TNF-alpha. In addition, upon ex vivo exposure to either myelin basic protein peptide 63-88 (MBP63-88) or to phytoaemagglutinin A, the numbers of IFN-gamma secreting splenocytes was also significantly reduced (ELISPOT analysis) in rhIL-6-treated rats as compared to PBS-treated controls. PMID- 11438172 TI - Theiler's virus-infected L-selectin-deficient mice have decreased infiltration of CD8(+) T lymphocytes in central nervous system but clear the virus. AB - Mice with targeted deletion of L-selectin gene (L-sel(-/-)) were used to investigate the role of adhesion molecule in immunologic responses following virus infection in the central nervous system (CNS). L-Sel(-/-) mice from a resistant H-2(b) genetic background and parental wild-type H-2(b) (C57BL/6) mice were infected with Theiler's murine encephalomyelitis virus (TMEV) intracerebrally and the kinetics of virus replication and infiltration of immune cells in the CNS determined. The levels of infectious TMEV, as measured by plaque assay at 3, 7, 14, and 28 days after infection were between 4 and 6 log(10) PFU of virus per gram of CNS tissues at days 3 and 7 post-infection, and then decreased to undetectable levels by day 14 after infection in both strains of mice. The L-sel(-/-) mice had decreased numbers of CD8(+) T lymphocytes (17.72%+/ 2.4) infiltrating into the CNS at 7 days post-infection when compared to wild type mice (31.02%+/-7.5). In addition, the L-sel(-/-) mice had significantly lower levels of TMEV-specific serum IgG resulting in lower virus neutralizing activity of the serum when compared to wild-type mice. However, the L-sel(-/-) mice had 2.5-fold increase in B lymphocytes in the CNS (8.29%+/-1.1) when compared to wild-type mice (3.2%+/-0.4). Taken together, these data indicate that L-selectin plays a role in recruitment of B and CD8(+) T lymphocytes into the CNS following virus infection, which, however, did not affect the ability of the mice to clear TMEV infection. PMID- 11438173 TI - A method for the selective depletion of perivascular and meningeal macrophages in the central nervous system. AB - The perivascular (PVM) and meningeal (MM) macrophages form a distinct population of resident CNS cells, selectively expressing the mature macrophage marker ED2 in the rat. In order to elucidate the role of the PVM and MM in rats during normal functioning of the brain and pathology, we have developed a strategy employing a single intraventricular injection of clodronate liposomes. This resulted in a complete depletion of the PVM and MM. Clodronate liposomes did not deplete the microglial cells. In other parts of the body, a temporal and mild depletion effect was observed, which was restored within 1 week. Detailed analysis of the elimination and repopulation kinetics of the PVM and MM revealed a slow repopulation of the CNS, starting at 14 days post depletion. This selective depletion method of the PVM and MM will enable us to get direct insight in their functions during normal and pathologic conditions of the CNS. PMID- 11438174 TI - Brain-derived gangliosides suppress the chronic relapsing-remitting experimental autoimmune encephalomyelitis in NOD mice induced with myelin oligodendrocyte glycoprotein peptide. AB - Chronic relapsing-remitting experimental autoimmune encephalomyelitis (CREAE) induced with myelin oligodendrocyte glycoprotein peptides 35-55 (MOG(35-55)) in NOD mice was successfully treated with brain-derived gangliosides (GA). The GA treatment suppressed the development and severity of CREAE, both clinically and histologically. Spleen cells from the GA-treated mice displayed markedly inhibited levels of MOG(35-55) specific proliferation and interferon-gamma production. Delayed-type hypersensitivity reactions to MOG(35-55) were suppressed by the GA treatment. GA modulate various T cell effector functions in CREAE and may be an effective therapeutic agent for autoimmune demyelinating diseases such as multiple sclerosis. PMID- 11438175 TI - Variability in immunohistochemistries of IgM M-proteins binding to sulfated glucuronyl paragloboside. AB - Serum IgMs from 4 of 12 patients with polyneuropathy and IgM M-proteins that bind to sulfated glucuronyl paragloboside (SGPG) strongly immunostained the human peripheral nerve myelin (group A), whereas those from the other eight patients strongly immunostained the cytoplasm of the Schwann cells surrounding the myelin sheath with only weak staining of the myelin (group B). Strong immunostaining of peripheral myelin by IgMs from group A patients may be due to the strong cross reactivities against P0 and peripheral myelin protein-22 (PMP-22), which are localized in compact myelin. Only three patients (all in group B) showed some response to the immunotherapies. Weak reactivities to P0 and and PMP-22 might indicate the possibility of improvement after the immunotherapies. PMID- 11438176 TI - In vivo gene expression revealed by cDNA arrays: the pattern in relapsing remitting multiple sclerosis patients compared with normal subjects. AB - OBJECTIVES: To use DNA arrays to identify differences in gene expression associated with relapsing-remitting (RR) MS. METHODS: Total RNA was isolated from monocyte depleted peripheral blood mononuclear cells of 15 RR MS patients and 15 age- and sex-matched controls. The RNA was reverse transcribed to radiolabeled cDNA and the resultant cDNA was used to probe a DNA array containing over 4000 named human genes. The binding of radiolabeled cDNA to the probes on the array was measured by phosphorimager. RESULTS: Of more than 4000 genes tested, only 34 were significantly different in RR-MS patients from controls. Of these, 25 were significantly increased and 9 significantly decreased in the RR MS patients. Twelve of these genes have inflammatory and/or immunological functions that could be relevant to the MS disease process. The potentially relevant genes that were elevated (15% to 28%) were P protein, LCK, cAMP responsive element modulator, IL 7 receptor, matrix metalloproteinase-19, M130 antigen, and peptidyl-prolyl isomerase. Those that were significantly decreased (15% to 35%) were SAS transmembrane 4 superfamily protein, STRL22 (C-C chemokine receptor 6), AFX protein, DNA fragmentation factor-45 and immunoglobulin gamma 3 (Gm marker). CONCLUSIONS: The RR-MS disease effect was relatively restricted and most of the mRNAs tested were not different from the normal controls. However, there were significant differences identified in the expression of a subset of mRNAs, including 13 with inflammatory/immune functions that could be relevant to MS. The systematic use of DNA arrays can provide insight into the dynamic cellular pathways involved in MS pathogenesis and its phenotypic heterogeneity. PMID- 11438177 TI - Investigation of serum response to PMP22, connexin 32 and P(0) in inflammatory neuropathies. AB - We investigated serological immune responses against three major peripheral nerve myelin constituents in patients with immune-mediated neuropathies. Connexin 32 (Cx32), myelin protein zero (MPZ, P(0)) and peripheral myelin protein 22 (PMP22) were produced in Chinese hamster ovary (CHO)-K1 cells. From a panel of 25 Guillain-Barre syndrome (GBS) and 24 chronic inflammatory demyelinating polyneuropathy (CIDP) patients, only two patients showed reactivity towards Cx32 and P(0). PMID- 11438178 TI - Expression and distribution of transcription factor NF-kappaB and inhibitor IkappaB in the inflamed peripheral nervous system. AB - The NF-kappaB family of transcription factors is critically involved in the immune response. The activity of these proteins is under strict control of an inhibitory molecule called IkappaB. The present study investigated the expression and distribution pattern of NF-kappaB and IkappaB in sural nerve biopsies obtained from patients with Guillain-Barre syndrome, chronic inflammatory demyelinating polyradiculoneuropathy, and various non-inflammatory neuropathies. In inflammatory demyelinating as well as non-inflammatory neuropathies, NF-kappaB was primarily expressed by macrophages, as determined by immunohistochemistry. IkappaB, however, could be localized to macrophages as well as T cells in inflammatory demyelinating neuropathies, whereas in non-inflammatory controls Schwann cells were found to be the primary cell type expressing this inhibitor. Quantitation of immunoreactivity revealed a statistically significant increase of NF-kappaB expression in inflammatory demyelinating cases compared to controls. Our results suggest an important function of the NF-kappaB pool in the genesis of inflammatory demyelination in the peripheral nervous system. PMID- 11438179 TI - Development of an ultrasensitive enzyme immunoassay for the determination of matrix metalloproteinase-9 (MMP-9) levels in normal human cerebrospinal fluid. AB - Determination of matrix metalloproteinase-9 (MMP-9) in human cerebrospinal fluid (CSF) to study blood-brain barrier impairment and immune cell migration in inflammatory neurological diseases recently became a matter of major interest. Regularly, MMP-9 was determined qualitatively or semi-quantitatively by zymography (gelatin gel electrophoresis) or quantitatively by enzyme immunoassay (EIA). As yet, it was not possible by either method to detect MMP-9 in CSF of controls (patients without pathologically increased CSF parameters). We developed an ultrasensitive two-side enzyme-linked immunosorbent assay (ELISA) which allows for the first time to measure reliably MMP-9 concentrations in CSF of controls. This ELISA uses a monoclonal as capture and a polyclonal as detector antibody. The detection limit of the assay is below 10 pg/ml and the assay range is 15-2000 pg/ml. Intra-assay precision is 2.5% for low and 3.7% for high, inter-assay precision is 11% for low and 10.7% for high values, respectively. The determination of the MMP-9 concentration in 50 control CSF gave the following results: range, 22-146 pg/ml; median, 76 pg/ml. The measurement of native and recombinant MMP-9 was carried out with three commercially available ELISAs, most widely employed in MMP-9 research, and compared to the newly developed one. All ELISAs recognize recombinant MMP-9 by factors of 5-20 less sensitively than native MMP-9. PMID- 11438180 TI - Association of a CA repeat polymorphism upstream of the Fas ligand gene with multiple sclerosis. AB - We have analyzed a CA repeat polymorphism localized 46-kb upstream of the Fas ligand gene in Spanish and American populations that include 139 healthy controls and a cohort of 177 unrelated relapsing and remitting multiple sclerosis (MS) patients. The MS patients consisted of two groups, one with a family history of MS and one without. The frequency of the 13 CA repeats (allele B) was lower (p=0.01) in MS patients than in controls, 0.45 and 0.55 respectively. The odds ratio (BB vs. AB/AA) for MS patients vs. healthy controls was 0.51 (95% CI 0.3 0.9; p=0.01). The odds ratio (BB vs. AB/AA) for MS patients extracted from multiply affected families vs. healthy controls was 0.22 (95% CI 0.07-0.62; p=0.002). The HLA DRB1*1501-DQB1*0602 haplotype is associated with B allele with a relative frequency higher than A allele (0.52 and 0.48 in patients vs. 0.68 and 0.32 in controls). The results suggest that chromosomes with B allele have a genetic background that reduces susceptibility to MS, particularly in the familial forms. PMID- 11438182 TI - Use of selective inhibitors and chromogenic substrates to differentiate bacteria based on toluene oxygenase activity. AB - In whole-cell studies, two alkynes, 1-pentyne and phenylacetylene, were selective, irreversible inhibitors of monooxygenase enzymes in catabolic pathways that permit growth of bacteria on toluene. 1-Pentyne selectively inhibited growth of Burkholderia cepacia G4 (toluene 2-monooxygenase [T2MO] pathway) and B. pickettii PKO1 (toluene 3-monooxygenase [T3MO] pathway) on toluene, but did not inhibit growth of bacteria expressing other pathways. In further studies with strain G4, chromogenic transformation of alpha,alpha,alpha-Trifluoro-m-cresol (TFC) was irreversibly inhibited by 1-pentyne, but the presence of phenol prevented this inhibition. Transformation of catechol by G4 was unaffected by 1 pentyne. With respect to the various pathways and bacteria tested, phenylacetylene selectively inhibited growth of Pseudomonas mendocina KR1 (toluene 4-monooxygenase [T4MO] pathway) on toluene, but not on p-cresol. An Escherichia coli transformant expressing T4MO transformed indole or naphthalene in chromogenic reactions, but not after exposure to phenylacetylene. The naphthalene reaction remained diminished in phenylacetylene-treated cells relative to untreated cells after phenylacetylene was removed, indicating irreversible inhibition.These techniques were used to differentiate toluene degrading isolates from an aquifer. Based on data generated with these indicators and inhibitors, along with results from Biolog analysis for sole carbon source oxidation, the groundwater isolates were assigned to eight separate groups, some of which apparently differ in their mode of toluene catabolism. PMID- 11438183 TI - The effect of fungal species on the fluorescent lectin test. AB - Fungal (mold) contamination is an important indicator of low-quality raw product used in food processing operations. Fluorescent-labeled lectins, specific for chitin, have been shown to be valuable for quantitative detection of mold in raw tomatoes. In this research, the response of individual fungal species to a rapid fluorescent lectin assay was investigated. Ten of the most common mold species were grown on two types of artificial broth media, and added to blended field tomatoes. The assay was conducted on each species, and linear regressions were developed, comparing the fluorescent lectin assay score with the fungal dry weight. The assay was able to detect all molds at sensitivities required for the tomato industry, and had high linearity (r2 ranging from 0.72 to 0.99) and low variability (standard error of calibration ranging from 20 to 116 microg of fungal biomass/ml of tomato juice) for individual species grown on V-8 juice broth. PMID- 11438184 TI - Integration of Raman microscopy, differential interference contrast microscopy, and attenuated total reflection Fourier transform infrared spectroscopy to investigate chlorhexidine spatial and temporal distribution in Candida albicans biofilms. AB - Two spectroscopic techniques, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and Raman microscopy (RM), were used to characterize transport of chlorhexidine digluconate (CHG) in Candida albicans (CA) biofilms. Different (volumetric) regions of the biofilm are sampled by these two vibrational spectroscopies making them complementary techniques. Simple mathematical models were developed to analyze ATR-FTIR and RM data to obtain an effective diffusion coefficient describing transport through CA biofilms. CA biofilms were composed primarily of yeast and hyphal forms, with some pseudohyphae. Upper regions of biofilms that had become confluent, (i.e., biofilms that completely covered the germanium (Ge) substratum) were composed primarily of a tangled mass of hyphae with openings between germtubes about 10 to 50 microm across. Quantitative analysis of ATR-FTIR kinetic data curves indicated that the effective diffusion coefficient for transport of CHG through confluent biofilms about 200-microm thick was reduced 0.1 to 0.3 times compared to the diffusion coefficient for CHG in water. Effective diffusion coefficients obtained from analysis of RM data were consistently higher than those indicated by ATR FTIR data suggesting that transport is more hindered in regions near the base of the biofilm than in the outer layers. Analysis of both ATR-FTIR and RM data obtained from thicker films indicated that adsorption of CHG to biofilm components was responsible for a substantial portion of the transport limitation imposed by the biofilm. Comparison of ATR-FTIR and RM data for both types of biofilms indicated that sites of CHG adsorption were more concentrated in the interfacial region than in the bulk biofilm. Comparison of results for ATR-FTIR and RM measurements suggests that these relatively thick CA biofilms can be modeled, for purposes of predicting transport, approximately as a homogeneous thin planar sheet. Thus, these biofilms offer a relatively tractable model system for initial investigations of the relation between antimicrobial transport and kinetics of antimicrobial action. PMID- 11438185 TI - A luciferase-based method for assessing chlorine-susceptibility of Mycobacterium avium. AB - A rapid and quantitative assay for the disinfection of the water-borne pathogen, Mycobacterium avium, was developed using firefly luciferase as a reporter gene. There was a correlation between the quantity of light produced and the number of colony-forming units. In chlorine-disinfection studies of a luciferase-carrying derivative of M. avium, there was a strong correlation (r2=0.96) between colony forming units and relative light units. It was discovered that chlorine was rapidly lost from suspensions containing 10(6) M. avium cells/ml. The luciferase based test can be used to rapidly measure susceptibility of M. avium to different disinfectants used in water treatment. PMID- 11438186 TI - Isolation and characterization of a temperature-sensitive generalized transducing bacteriophage for Vibrio cholerae. AB - CP-T1 is the only described generalized transducing bacteriophage for the intestinal pathogen Vibrio cholerae, yet many of its basic biological parameters remain unknown. Due to low frequencies of transduction and pseudolysogen formation, CP-T1 has not been widely used as a genetic tool. To overcome these limitations, we have isolated a conditional mutant of CP-T1 that exhibits temperature-sensitive plaque formation. Several biological properties of CP-T1ts were determined, including its restrictive temperature, adsorbance profile to host cells, burst time, and burst size. Based on these properties, an optimized transduction protocol was designed which resulted in several fold higher transduction frequencies for a variety of genetic markers from a number of chromosomal loci. Generalized transduction was also demonstrated between classical and E1 Tor biotype strains of V. cholerae. PMID- 11438187 TI - Determination of the growth stages of Lactobacillus paracasei subsp. paracasei M3 from Bulgarian yellow cheese by electroconductivity. AB - The growth curve and the bacteriocin production by Lactobacillus paracasei subsp. paracasei M3-a strain isolated from Bulgarian yellow cheese, was studied using electroconductivity measurements in the culture medium (MRS). The bacteriocin produced by this bacterium inhibits strains belonging to the Candida species as well as Bacillus subtilis ATCC 6633. The use of the direct impedance technique coupled with statistical treatment of the results for the strain's growth curve allowed the detection of the growth phase-related bacteriocin production. Definition of the average particle size and aggregation estimations for the cells of the bacteriocin-producing strain was also possible. The comparison of results of impedance measurements with results obtained by classic microbiological methods and microscopic observations confirms the good correlation between these methods. In addition, the advantages of an impedance method such as rapidity, simplicity and higher sensitivity are highlighted. PMID- 11438188 TI - New immunoassay for the detection of Helicobacter pylori infection compared with urease test, 13C breath test and histology: validation in the primary care setting. AB - Helicobacter pylori plays a major role in peptic ulcer disease and, as a result, testing for H. pylori infection among patients with dyspepsia has often been advocated. The aim of the study was to determine the diagnostic accuracy, the analytical performance, and optimal cut-off point of a new serological assay, the Pyloriset EIA-G III for the detection of H. pylori infection in the primary care setting. For 113 primary care patients with dyspepsia urea breath test, CLO test, histology and serology tests were performed. Diagnostic accuracy of the Pyloriset EIA-G III was evaluated against a reference standard of a carbon urea breath test (CUBT), CLO test and histology (from gastric biopsies). Precision, linearity and correlation of the serological assay with the CUBT and former Pyloriset were also determined. At the optimal cut-off level of 40 U/ml, the positive predictive value was 92.1%, negative predictive value 96.3%, sensitivity 87.5%, and specificity 93.9%. The within-run precision was high. The recovery data were good. The correlation of both CUBT and the former Pyloriset EIA-G and the Pyloriset EIA-G III was high. At the cut-off level of 40 U/ml, the new Pyloriset EIA-G III is a reliable method to detect H. pylori infection in the primary care setting. PMID- 11438189 TI - Evaluation of real-time quantitative PCR for identification and quantification of Chlamydia pneumoniae by comparison with immunohistochemistry. AB - Chlamydia pneumoniae is a common cause of community-acquired pneumonia and it has been associated with atherosclerosis. C. pneumoniae has usually been diagnosed by serology using a microimmunofluorescence test, but more recently polymerase chain reaction (PCR) has been viewed as an advantageous alternative. We developed a quantitative real-time PCR for detection of C. pneumoniae. Primers were targeted for the pmp4 gene, and the PCR fragment was detected real-time with a fluorescence resonance energy transfer probe set using a LightCycler instrument. The PCR was used on DNA released from 50 microm sections of paraffin-embedded formalin-fixed lung tissue from experimentally infected mice. Thereby, the number of C. pneumoniae genomes was determined. To our knowledge this is the first time quantification of C. pneumoniae DNA has been attempted on paraffin-embedded formalin-fixed tissue. C. pneumoniae-specific immunohistochemistry (IHC) was done on 5 microm sections adjacent to the sections used in PCR, and the number of inclusions were counted in each section. Good correlation was found when comparing results from PCR and IHC, which is in contrast to many previous studies. PMID- 11438190 TI - Colorimetric determination of active alpha-glucoside transport in Saccharomyces cerevisiae. AB - Fermentation of alpha-glucosides (maltose, maltotriose) by Saccharomyces cerevisiae cells is a critical phase in the processes of brewing and breadmaking. Utilization of alpha-glucosides requires the active transport of the sugar across the cell membrane and, subsequently, its hydrolysis by cytoplasmic glucosidases. Although transport activities are usually assayed using radiolabeled substrates, we have developed a simple, cheap and reliable colorimetric assay for the determination of alpha-glucoside uptake using p-nitrophenyl-alpha-D glucopyranoside (pNPalphaG) as substrate. Our results show that pNPalphaG is actively transported by S. cerevisiae cells by a H+-symport mechanism, which depends on the electrochemical proton gradient across the plasma membrane. pNPalphaG uptake is mediated by the AGT1 alpha-glucoside permease, which has a high affinity (Km=3 mM) for this chromogenic substrate. This simple colorimetric uptake assay can be used to analyze the expression and regulation of the AGT1 permease in S. cerevisiae cells. PMID- 11438191 TI - Potential problems with fluorescein diacetate assays of cell viability when testing natural products for antimicrobial activity. AB - There are two potential problems in the use of fluorescein diacetate (FDA) as a measure of cell viability. The first is the hydrolysis of FDA to fluorescein in the absence of live cells and the second is the quenching of fluorescence by assay solutions. We show that common media components such as tryptone, peptone and yeast extract all promote hydrolysis of FDA in the absence of live cells, as do Tris-HCl and sodium phosphate buffers. As a consequence, various microbiological media promote hydrolysis of FDA in the absence of live cells. Different media were also shown to reduce the amount of visible fluorescence of fluorescein. Diluting the medium decreases the background hydrolysis of FDA as well as increases the amount of visible fluorescence. Both problems should be considered when using FDA as an indicator of cell viability when testing natural products for antimicrobial activity. PMID- 11438192 TI - Endothelial dysfunction, atherosclerosis and cyclosporine in renal transplantation. PMID- 11438193 TI - Bone marrow aspiration, a dangerous procedure? PMID- 11438194 TI - Cyclosporine is associated with endothelial dysfunction but not with platelet activation in renal transplantation. AB - BACKGROUND: Cyclosporine (CsA) is associated with thrombotic micro-angiopathy and endothelial dysfunction. Markers of endothelial dysfunction may serve to identify patients at risk for development of vascular injury. In this study we measured von Willebrand Factor (vWF) and sP-selectin as possible markers for endothelial dysfunction in renal transplant recipients at different concentrations of CsA. Because sP-selectin can also be derived from platelets an additional in vitro study was performed to study the potential effect of CsA on the expression of P selectin on platelet surface, while the effects of CsA on the interaction of platelets with Endothelial Cell Matrix (ECM) were studied under flow conditions in a perfusion chamber model. METHODS: CsA was stepwisely replaced by mycophenolate mofetil (MMF) in 15 renal transplant recipients (more than 6 months after transplantation). VWF and sP-selectin were measured at normal CsA (median trough level 130 microg/l), low CsA (trough level 45 microg/l) and after stopping CsA. MMF 2 g daily was added while lowering and stopping CsA. Platelet activation was investigated by measurement of P-selectin on platelet surface by flow cytometry (FACS), after incubation with CsA (0, 2, 20 and 200 mg/l) in vitro and after perfusion of whole blood over ECM with CsA (0 or 2 mg/l, peak levels). RESULTS: Stepwise withdrawal of CsA gave a dose-related decrease of both vWF and sP-selectin, suggesting reversible endothelial dysfunction. FACS showed no expression of P-selectin on platelets by CsA. Also perfusion studies over ECM demonstrated no platelet activation by CsA but even inhibition of platelet adhesion and aggregation. CONCLUSIONS: The use of CsA is not accompanied by platelet activation. However endothelial dysfunction induced by CsA does occur as reflected by increased vWF and sP-selectin. (See Editorial p. 1). PMID- 11438195 TI - High fat intake in hyperlipidaemic patients is related to male gender, smoking, alcohol intake and obesity. AB - BACKGROUND: In individuals at high cardiovascular risk, such as patients with hyperlipidaemia, low dietary fat intake is used to reduce this risk. The aim of the present study was to identify determinants of (saturated) fat intake in hyperlipidaemic patients. METHODS: Cross sectional study in a lipid clinic of a tertiary referral centre. A total of 1169 patients (714 males and 455 females) with hyperlipidemia were studied. Food frequency questionnaires were present of 1026 patients. In 615 patients a detailed diet analysis was performed. The main outcomes measures were determinants of fat intake, indicated by a regression coefficient (beta-coefficient). RESULTS: The following variables were independently related to fat intake: present smoking (beta-coefficient 3.7), male gender ((beta 1.6), familial hypercholesterolemia (beta -1.6), alcohol (beta 0.6 per glass of alcohol), body mass index (beta 0.6). No interaction between gender and smoking or between gender and alcohol intake was observed in relation to fat intake. The percentage of energy from fats were higher in males than in females, 34.2+/-8.3% and 31.7+/-8.3%, respectively (P<0.001). The higher total and saturated fat intake in males is due to a larger consumption of cheese, meat products, bread and potato products. Women had a higher relative intake of carbohydrate 48.5+/-8.7% versus 46.5+/-8.8% in males (P<0.05), due to a relatively higher intake of fruit, milk products and pastry and biscuits. CONCLUSIONS: A specific gender-oriented approach may improve the results of dietary counselling of hyperlipidaemic patients. PMID- 11438196 TI - Functional bowel symptoms in a general Dutch population and associations with common stimulants. AB - BACKGROUND: Functional gastrointestinal symptoms are often felt to be related to the use of alcohol, coffee and smoking. METHODS: A random sample of 500 adults was interviewed by telephone about their gastrointestinal symptoms and the use of alcohol, coffee and smoking. Dyspepsia and irritable bowel syndrome (IBS) were defined using common, internationally used criteria. RESULTS: Of those invited, 85.4% agreed to participate (43.5% male). Of the participants 21.4% had gastrointestinal symptoms >6 times/last year, 13.8% had dyspepsia, and 5.8% had IBS, the latter being more common among women. Of the men, 83.9% reported alcohol consumption, 92.0% drank coffee, and 52.2% smoked. Of the women 62.4% drank alcohol, 88.1% drank coffee and 29.8% smoked. Use of alcohol or coffee was not related to dyspepsia. Current smokers had a 2.3-fold increased risk (P=0.02) and former smokers a 2.7 increased risk (P=0.009) for dyspepsia compared to never smokers. No association between IBS and use of alcohol, coffee or smoking could be demonstrated. CONCLUSIONS: The prevalence of dyspepsia (13.8%) and of IBS (5.8%) in a general Dutch adult population appears to be lower than are reported in other countries. Use of alcohol and coffee was not associated with functional bowel symptoms. Former and current smoking were strongly related to dyspepsia. PMID- 11438197 TI - The association of Sweet's syndrome with sarcoidosis. AB - We report a 43-year-old woman with biopsy proven Sweet's syndrome with massive mediastinal lymphadenopathy. No other clinical signs compatible with sarcoidosis were present. Mediastinoscopy was performed to establish a diagnosis. Histological evaluation revealed non-caseating granulomas compatible with sarcoidosis. We review the association of Sweet's syndrome and (haematological) malignancies, and eight other cases of Sweet's syndrome and sarcoidosis. All but one presented with erythema nodosum, making the diagnosis of acute sarcoidosis (Lofgren's syndrome) more likely than malignancy. PMID- 11438198 TI - Eosinophilic tubulo-interstitial nephritis associated with iridocyclitis and thyreoiditis. AB - A patient is described with the tubulointerstitial nephritis with uveitis syndrome. The diagnosis can be difficult since it has to be differentiated from sarcoidosis, or infections like tuberculosis and toxoplasmosis. Our patient showed prompt recovery of fever, ocular symptoms and renal function after starting corticosteroids. PMID- 11438199 TI - Cardiac tamponade following sternal puncture in two patients. AB - Sternal puncture for harvesting bone marrow is a procedure with potentially lethal complications. Both the occurrence of pneumothorax and sternal fracture are reported. We present two patients with a cardiac tamponade following elective sternal puncture. (See Editorial p. 4). PMID- 11438201 TI - Meeting summary: International Symposium and Workshop on Hematopoietic Stem Cells III, University of Tubingen, Germany, September 14-16, 2000. PMID- 11438202 TI - Interleukin-3 and stem cell factor modulate cell cycle regulatory factors in mast cells: negative regulation of p27Kip1 in proliferation of mast cells induced by interleukin-3 but not stem cell factor. AB - OBJECTIVE: Interleukin-3 (IL-3) and stem cell factor (SCF) are able to promote survival and proliferation of mast cells. However, the precise signal transduction cascades leading to mast cell proliferation are not clearly understood. Thus, we sought to define the mechanism of mast cell proliferation induced by IL-3 and SCF. MATERIALS AND METHODS: We treated murine bone marrow derived cultured mast cells (BMCMC) with recombinant IL-3 (rIL-3) or recombinant SCF (rSCF) and examined the effects of rIL-3 and rSCF on cell cycle regulatory factors. RESULTS: Both rIL-3 and rSCF suppressed apoptosis of BMCMC. rSCF induced great proliferation of BMCMC with elevation of the proportions of cells in S and G2/M phases, whereas most BMCMC incubated with rIL-3 were arrested in the G1 phase. The G1/S phase transition is initiated by phosphorylated retinoblastoma protein (pRb), which was prominent in cells stimulated with rSCF. In contrast, rIL-3 relatively increased a dephosphorylated form of pRb in BMCMC. Compared with rIL-3, rSCF induced greater expression of cyclin-dependent kinase (CDK) 2 and CDK4, which are able to phosphorylate pRb, and cyclin D3, a partner of CDK4. BMCMC treated with rIL-3 contained a high amount of a CDK inhibitor p27Kip1 that was suppressed by pretreatment with Ro31-7549, a protein kinase C inhibitor, whereas rSCF induced weak expression of p27Kip1 in BMCMC. CONCLUSION: The results suggest that IL-3 and SCF exert their respective mitogenic effects on mast cells by modulating the expression of pRb, CDK, cyclin, and p27Kip1. PMID- 11438203 TI - Interleukin-6 is a major effector molecule of short-term G-CSF treatment inducing bone metabolism and an acute-phase response. AB - OBJECTIVE: The biological steps leading to hematopoietic progenitor cell (HPC) mobilization from the bone marrow to the peripheral blood compartment during G CSF treatment are still poorly defined. In this study, we investigated G-CSF mediated secretion of cytokines as potential mediators. MATERIALS AND METHODS: Plasma and urine samples from G-CSF-mobilized donors for HPC transplantation were collected before and during mobilization therapy. Interleukin-6 (IL-6), alkaline phosphatase (ALP), bone-specific ALP (bone ALP), C-reactive protein (CRP), lipopolysaccharide binding protein (LPB), CD34+ cells and urinary deoxypyridinoline (DPD) concentrations were measured and statistically correlated. RESULTS: IL-6 was highly elevated at days 4 and 5 of G-CSF treatment. G-CSF administration led to elevation of IL-6 parallel to the appearance of CD34+ cells in the peripheral blood. Major metabolic changes such as high bone ALP plasma concentration and urinary excretion of deoxypyridinoline (DPD), indicating stimulation of bone metabolism, were observed. Elevated concentrations of CRP and LPB indicated an acute-phase response. Furthermore, CRP concentrations correlated significantly with the percentage of mobilized CD34+ cells. CONCLUSION: Based on these findings, we propose IL-6 as a major physiological effector molecule of G CSF treatment that induces bone metabolism and an acute-phase reaction along with mobilization of CD34+ cells in the peripheral blood. IL-6 may be responsible for osteopenia observed during short- and long-term G-CSF treatment. These observations may also have implications for G-CSF treatment of patients with plasmocytoma. PMID- 11438204 TI - gp130-stimulating designer cytokine Hyper-interleukin-6 synergizes with murine stroma for long-term survival of primitive human hematopoietic progenitor cells. AB - OBJECTIVE: Experimental strategies for ex vivo expansion of human hematopoietic stem/progenitor cells involve the use of exogenous cytokines as well as direct interaction with stromal elements. We examined the use of the interleukin 6/soluble interleukin-6 receptor (IL-6/sIL-6R) fusion protein Hyper-IL-6 (H-IL 6), which interacts directly with gp130, in conjunction with stromal support for expansion of human progenitors. MATERIALS AND METHODS: Peripheral blood CD34+ cells were cultured on the murine stromal cell line FBMD1 or in suspension for up to 28 days with different cytokines. Cells were evaluated at various time points for phenotype, proliferative and clonogenic capacity, and long-term hematopoietic activity. RESULTS: The combination of Flt3 ligand and H-IL-6 was markedly more effective than Flt3 ligand and IL-6/sIL-6R for expansion of CD34+ cells in suspension culture and on FBMD1. Addition of kit ligand but not thrombopoietin to Flt3 ligand and H-IL-6 significantly augmented proliferation and enhanced colony formation three-fold. However, long-term cobblestone area-forming cell assays indicated that although multipotent progenitors were maintained up to 21 days on FBMD1 in the presence of Flt3 ligand alone and were amplified three-fold by addition of H-IL-6, CD34+ cells cultured in the absence of stromal support rapidly lost their cobblestone area-forming cell potential. Immunophenotyping revealed that stromal support prevented up-regulation of IL-6R on CD34+ cells, which was induced within 3 days in stroma-free cultures and was enhanced in the presence of kit ligand. Delayed addition of H-IL-6 to the cultures resulted in reduced proliferation and colony-forming unit potential. CONCLUSION: H-IL-6 synergizes with stromal elements to effectively enhance proliferation and maintenance of primitive hematopoietic progenitors under prolonged ex vivo culture conditions. PMID- 11438205 TI - Stem cell factor protects c-kit+ human primary erythroid cells from apoptosis. AB - OBJECTIVE: It has been reported that stem cell factor (SCF) promotes cell survival in primary cultured human erythroid colony-forming cells (ECFC). Given the heterogeneous nature of ECFC, which may affect interpretation of the data, we purified c-kit+ ECFC and investigated the specificity and mechanisms of the anti apoptotic effects of SCF on these cells. MATERIALS AND METHODS: Glycophorin A+ (GPA+) c-kit+ cells were purified from primary cultured ECFC derived from purified human CD34+ cells. The GPA+c-kit- and nonerythroid cells were generated from the same CD34+ cells. Apoptosis of ECFC was investigated in the absence or presence of SCF and erythropoietin (EPO) in serum-free medium. DNA fragmentation was measured with enzyme linked immunosorbent assay for oligonucleosome-sized DNA, gel electrophoresis, and annexin V labeling. Characterization of expanded cells and enriched cells was performed using multiparameter flow cytometry. For Akt assay, cells were lysed and the cleared lysates subjected to SDS-PAGE followed by Western blotting. RESULTS: In GPA+c-kit+ cells, deprivation of cytokine caused rapid DNA fragmentation within 4 hours that reached a maximum at 6 hours. This was partially but clearly prevented by SCF or EPO. In contrast, no significant DNA fragmentation was seen in GPA+c-kit- and nonerythroid cells within 24 hours. PP2, a specific Src family kinase inhibitor, but not its inactive analogue PP3, reversed the anti-apoptotic effects of SCF. PP2 also inhibited SCF-induced phosphorylation of Akt. CONCLUSION: These data indicate that SCF protects purified human GPA+c-kit+ cells from apoptosis and suggest that kit-mediated Src kinase activation is involved in Akt activation and cell survival. PMID- 11438206 TI - Functional analysis of patient-derived mutations in the Fanconi anemia gene, FANCG/XRCC9. AB - OBJECTIVE: Fanconi anemia (FA) is an autosomal-recessive cancer susceptibility syndrome with seven complementation groups. Six of the FA genes have been cloned (corresponding to subtypes A, C, D2, E, F, and G) and the encoded proteins interact in a common pathway. Patient-derived mutations in FA genes have been helpful in delineating functional domains of FA proteins. The purpose of this work was to subtype FA patient-derived cell lines in our repository and to identify FA gene mutations. METHODS: We subtyped 62 FA patients as type A, G, C, or non-ACG by using a combination of retroviral gene transfer and immunoblot analysis. Among these FA patients, we identified six FA-G patients for further analysis. We used a strategy involving amplification of FANCG/XRCC9 exons and direct sequencing to identify novel FANCG mutations in cell lines derived from these FA-G patients. We functionally analyzed FANCG mutant alleles by transducing the corresponding cDNAs into a known FA-G indicator cell line and scoring correction of MMC sensitivity. RESULTS: Our results demonstrate a wide range of mutations in the FANCG gene (splice, nonsense, and missense mutations). Based on this mutational screen, a carboxy terminal functional domain of the FANCG protein appears to be required for complementation of FA-G cells and for normal assembly of the FANCA/FANCG/FANCC protein complex. CONCLUSION: The identification of patient-derived mutant alleles of FA genes can provide important insights to the function of FA proteins. FA subtyping is also a necessary precondition for gene therapy. PMID- 11438207 TI - Expression of Jagged1 gene in macrophages and its regulation by hematopoietic growth factors. AB - OBJECTIVE: Serrate/Jagged and Delta are cell surface ligands for Notch receptors that may influence hematopoietic cell fate decisions and are known to be expressed in bone marrow stromal cells. In a series of screenings of cDNAs constructed by a cDNA library subtraction technique, we identified Jagged1, one of the Notch ligands, as a gene up-regulated by macrophage colony-stimulating factor (M-CSF) in bone marrow macrophages. Therefore, we compared stromal cells and macrophages for expression of Notch ligands including Jagged1 and analyzed the regulation of their expression by cytokines. MATERIALS AND METHODS: Murine bone marrow macrophages were prepared by culturing femoral bone marrow cells with M-CSF. Primary bone marrow fibroblastic stromal cells were prepared by a culture system that we recently developed. The expression of Notch ligands was analyzed by either Northern blot analysis or reverse transcriptase polymerase chain reaction. RESULTS: The bone marrow macrophages expressed Jagged1 but not Jagged2 and Delta1 at a level that was detectable by Northern blot analysis. Expression of the Jagged1 gene was markedly up-regulated by growth factors for the cells, i.e., M-CSF, granulocyte-macrophage colony-stimulating factor, and interleukin-3. Expression of Jagged2 and Delta1 seldom was affected by the stimuli. The primary bone marrow fibroblastic stromal cells, and murine stromal cell lines, such as PA6 and ST2, also expressed Jagged1 transcript, at levels comparable to the steady-state level in macrophages. However, expression of the Jagged1 gene was little affected when these cells were stimulated with fibroblastic growth factor and platelet-derived growth factor. CONCLUSIONS: We demonstrated that bone marrow macrophages as well as stromal cells constitutively produced Jagged1 and that the expression was markedly up-regulated by hematopoietic growth factors, M-CSF, granulocyte-macrophage colony-stimulating factor, and interleukin-3. The results highlight the involvement of macrophages and these growth factors in hematopoietic cell fate decisions via the production of Jagged1. PMID- 11438208 TI - Inhibition of myeloid differentiation by Hoxa9, Hoxb8, and Meis homeobox genes. AB - OBJECTIVE: The homeobox gene Hoxb8 is activated in the murine myelomonocytic cell line WEHI-3B as a result of intracisternal A particle integration. Cooperative activation between Hoxa9 and Meis1 is induced by retroviral integration in BXH2 murine myeloid leukemias and the myeloid leukemia cell line M1. The present study was conducted to examine possible Meis gene activation and cooperative DNA binding of homeobox proteins in WEHI-3B and to reveal the specific role of Hox and Meis genes in myeloid differentiation. MATERIALS AND METHODS: Northern blot analysis and reverse transcriptase polymerase chain reaction were performed to examine homeobox genes expression. Electrophoretic mobility shift assay was performed to evaluate DNA binding of homeobox proteins. Myeloid differentiation of 32Dcl3 was induced by granulocyte colony-stimulating factor. RESULTS: Meis2 was coactivated with Hoxb8 in WEHI-3B cells. DNA-protein complexes including Hox, Meis, and Pbx were observed in WEHI-3B and 32Dcl3. Expression and the DNA-binding complex of Hoxa9, Hoxb8, Meis1, and Meis2 were down-regulated during myeloid differentiation of 32Dcl3 cells. Enforced expression of Hox or Meis genes inhibited myeloid differentiation of 32Dcl3. CONCLUSION: The results indicate that Meis2 is an important Meis gene for myeloid leukemogenesis and that Hox and Meis are important genes for myeloid leukemogenesis through differentiation block. PMID- 11438209 TI - Novel mutation in the PML/RARalpha chimeric gene exhibits dramatically decreased ligand-binding activity and confers acquired resistance to retinoic acid in acute promyelocytic leukemia. AB - OBJECTIVE: All-trans retinoic acid (RA) resistance in acute promyelocytic leukemia (APL) has been a serious clinical problem in differentiation-inducing therapy. However, the mechanisms underlying acquired RA resistance in APL patients are not well understood. MATERIALS AND METHODS: We recently established a spontaneous RA-resistant APL cell line (UF-1) from a patient and used this cell line as an excellent in vitro model for RA-resistant clinical situations. We investigated the structural and functional abnormalities of chimeric PML/RARalpha gene in UF-1 cells and preserved materials from the original patient. RESULTS: A novel point mutation was detected in the ligand-binding (E) domain of the RARalpha portion of the PML/RARalpha gene in UF-1 cells. This mutation resulted in amino acid substitution of Arg611 (CGG) for Trp611 (TGG) in the short-form PML/RARalpha protein, which corresponded to Arg276 in wild-type RARalpha. Importantly, the same mutation was also detected in the preserved materials from the original patient. COS-1 cells were transiently transfected with cDNA encoding wild-type and mutant PML/RARalpha constructed by site-directed mutagenesis and performed RA-binding assay. Interestingly, RA-binding activity was dramatically decreased in the mutant PML/RARalpha compared with that of the wild-type chimeric protein, suggesting that this single amino acid substitution is critical for RA binding. CONCLUSIONS: These results strongly suggest that a novel point mutation in the ligand-binding domain of the RARalpha portion (Arg611) of the chimeric PML/RARalpha gene decreased sensitivity to all-trans RA. We conclude that acquisition of the PML/RARalpha mutation is one possible mechanism for development of RA resistance in patients with APL in vivo. PMID- 11438210 TI - Oxygen tension modulates the expression of cytokine receptors, transcription factors, and lineage-specific markers in cultured human megakaryocytes. AB - OBJECTIVE: We have recently reported that 20% O2 significantly enhances total megakaryocyte (Mk) number, polyploidy, and proplatelet formation compared to 5% O2 in culture. In order to further elucidate the regulatory role of pO2 on megakaryocytopoiesis, we conducted a kinetic study of the expression of surface markers CD41a and CD42a; receptors for thrombopoietin (TPO), interleukin-3 (IL 3), and Flt3-ligand; the glutamate receptor of the N-methyl-D-aspartate subtype 1 (NMDAR1); and transcription factors GATA-1, NF-E2, and E2F-1. MATERIALS AND METHODS: Mks were generated from mobilized peripheral blood (PB) CD34+ cells from normal donors in serum-free medium with TPO, IL-3, and Flt3-ligand at 20% and 5% O2. Quantitative assessment of Mk surface receptors and nuclear transcription factors was performed using multiparameter flow cytometry. mRNA levels of the nuclear transcription factors GATA-1 and NF-E2 were evaluated using RT-PCR. RESULTS: The proportions of cells expressing the early Mk marker CD41a and the late Mk marker CD42a at day 15 were 4 and 5 times higher, respectively, at 20% O2. CD41a and CD42a protein levels per cell were also higher at 20% O2. After day 5, c-Mpl (TPO receptor) generally followed similar kinetics as CD41a. The proportion of IL-3 receptor (IL-3R)++ Mks at day 5 was 1.5 times higher at 5% O2. The NMDAR1 protein previously known to be expressed by neuronal cells has recently been identified in Mks. NMDAR1 and the transcription factors were studied on days 6, 9, and 11. NMDAR1 was expressed at a 1.5- to 1.8-fold higher level at 5% O2. Twenty percent O2 supported higher expression of the Mk-early and -late-maturation-specific transcription factors GATA-1 (1.2- to 2.2-fold higher) and NF-E2 (1.1- to 2.8-fold higher). This was consistent with RT-PCR data indicating the presence of higher levels of GATA-1 and NF-E2 mRNA at 20% O2. E2F 1, a ubiquitously expressed cell cycle transcription factor, was expressed at a 1.5-fold higher level at 20% O2 on day 6, but this difference did not persist by day 9. CONCLUSION: These findings demonstrate that cytokine receptors c-Mpl and IL-3R, and Mk differentiation-specific surface receptors CD41a, CD42a, and NMDAR1, are significantly modulated by pO2, and suggest that one of the mechanisms of enhanced maturation at 20% O2 may involve regulation of transcription factors GATA-1 and NF-E2. PMID- 11438211 TI - Differences in sulfation patterns of heparan sulfate derived from human bone marrow and umbilical vein endothelial cells. AB - OBJECTIVE: Heparan sulfates (HS), the polysaccharide side chains of HS proteoglycans, differ in structure and composition of sulfated domains among various tissue types, resulting in selective protein binding. HS proteoglycans on bone marrow endothelial cells (BMEC) could contribute to tissue specificity of the bone marrow endothelium and play a role in the presentation of chemokines such as stromal cell-derived factor-1 (SDF-1) and adhesion of hematopoietic progenitor cells after stem cell transplantations. We characterized differences in HS structure and SDF-1 binding between BMEC and human umbilical vein endothelial cells (HUVEC). MATERIALS AND METHODS: Expression of HS proteoglycans on human bone marrow microvessels was investigated by immunohistochemical staining. Comparison of three human BMEC cell lines with HUVEC and an HUVEC cell line was studied by flow cytometry using antibodies against different epitopes of the HS polysaccharide chain. HS proteoglycans were biochemically characterized after isolation from metabolically labeled cultures of the BMEC cell line 4LHBMEC and HUVEC. Binding of radiolabeled SDF-1 to 4LHBMEC and HUVEC and competition with heparins were investigated. RESULTS: Bone marrow microvessels constitutively expressed HS proteoglycans. Flow cytometric experiments showed differences in HS chain composition between BMEC and HUVEC. Biochemical characterization revealed more O-sulfation of the N-sulfated domains present in cell-associated HS glycosaminoglycans in 4LHBMEC compared to HUVEC. Binding experiments showed that 4LHBMEC bound more 125[I]-SDF-1 per cell than HUVEC. This could be inhibited largely by heparin and O-sulfated heparin and to a lesser extent by N-sulfated heparin. CONCLUSIONS: Cellular HS from BMEC differs in composition from HUVEC. We postulate that the presence of highly sulfated domains in the HS chains from BMEC contributes to tissue specificity of bone marrow endothelium in which HS may be involved in SDF-1 presentation and adhesion of hematopoietic progenitor cells. PMID- 11438212 TI - The homeodomain gene Pitx2 is expressed in primitive hematopoietic stem/progenitor cells but not in their differentiated progeny. AB - OBJECTIVE: Hematopoietic stem cells (HSCs) represent a rare and incompletely characterized fraction of marrow cells that are capable of both self-renewal and differentiation into all of the mature cells in the peripheral blood. We undertook to identify genes expressed preferentially by HSCs as an initial step toward better understanding the molecular mechanisms that underlie HSC behavior. METHODS: We modified the representational difference analysis technique to isolate gene fragments present in amplified cDNA prepared from highly purified murine hematopoietic stem/progenitor cells (Lin(-)/Hoechst(low)/rhodamine(low)) and absent (or much less abundant) in amplified cDNA prepared from lineage committed marrow cells. We went on to use one potentially important gene fragment that we isolated in this way, to screen a cDNA library prepared from these cells and to characterize the pattern of expression of the gene in hematopoietic and other cells. RESULTS: We isolated a fragment of the homeobox transcription factor Pitx2 from amplified cDNA prepared from murine hematopoietic stem/progenitor cells. From a cDNA library prepared from these cells, a full-length cDNA was isolated that corresponds to one of the three known isoforms of Pitx2 (Pitx2c). Pitx2c is expressed in murine embryonic stem (ES) cells and in hematopoietic stem/progenitor cells but not in more differentiated hematopoietic cells or in a large panel of established murine hematopoietic cell lines. Pitx2c expression was not detected after 48 hours of in vitro cytokine stimulation of hematopoietic stem/progenitor cells. CONCLUSIONS: Pitx2c is expressed in hematopoietic stem/progenitor cells but not in their differentiated progeny. The pattern of expression of Pitx2c in primitive hematopoietic stem/progenitor cells suggests that it may play a role in hematopoietic stem-cell biology. PMID- 11438213 TI - Cyclosporin A effects during primary and secondary activation of human umbilical cord blood T lymphocytes. AB - OBJECTIVE: Cyclosporin A (CsA), effective in prophylaxis and treatment of graft vs-host disease (GVHD) after human allogeneic transplantation, blunts T-cell responses by inhibiting nuclear factor of activated T cells-1 (NFAT1) activation. This laboratory has shown that NFAT1 protein expression is severely reduced in human UCB (umbilical cord blood) T cells. Since UCB is increasingly used as a hematopoietic stem cell source in allogeneic transplantation, it is important to determine whether CsA sensitivity in UCB differs from that of adult T cells. METHODS: Surface flow cytometric analysis, intracellular cytokine staining, flow cytometric analysis of cell death, and thymidine incorporation were used in this study to determine T-cell activation and effector functions during primary and secondary stimulation in the presence of CsA. RESULTS: Although we observed differential CsA sensitivity of T-cell activation marker (CD69, CD45RO, CD25) upregulation comparing UCB and adult, we did not observe any significant difference in CsA sensitivity of T-cell effector functions. Importantly, we observed reduced IFN-gamma and TNF-alpha expression in UCB T cells both in primary and secondary stimulation, as well as increased rates of activation induced cell death (AICD). CONCLUSION: Thus, our studies do not support the previous hypothesis that reduced GVHD observed after UCB transplantation is attributable to increased CsA sensitivity of UCB T cells. Rather, reduced UCB T cell cytokine production and increased AICD may be important cellular mechanisms underlying these favorable rates of GVHD in UCB transplant recipients. PMID- 11438214 TI - Human CD34+ cell preparations contain over 100-fold greater NOD/SCID mouse engrafting capacity than do CD34- cell preparations. AB - OBJECTIVE: The CD34 cell surface marker is used widely for stem/progenitor cell isolation. Since several recent studies reported that CD34(-) cells also have in vivo engrafting capacity, we quantitatively compared the engraftment potential of CD34(+) vs CD34(-) cell preparations from normal human placental/umbilical cord blood (CB), bone marrow (BM), and mobilized peripheral blood (PBSC) specimens, using the nonobese diabetic/severe combined immunodeficient (NOD/SCID) mouse model. METHODS: CD34(+) and CD34(-) cell preparations were purified by four different approaches in 14 individual experiments involving 293 transplanted NOD/SCID mice. In most experiments, CD34(+) cells were depleted twice (CD34(=)) in order to obtain efficient depletion of CD34(+) cells from the CD34(-) cell preparations. RESULTS: Dose-dependent levels of human hematopoietic cells were observed after transplantation of CD34(+) cell preparations. To rigorously assess the complementary CD34(-) cell preparations, cell doses 10- to 1000-fold higher than the minimum dose of the CD34(+) cell preparations necessary for engraftment were transplanted. Nevertheless, of 125 NOD/SCID mice transplanted with CD34(-) cell preparations purified from the same starting cells, only six mice had detectable human hematopoiesis, by flow cytometric or PCR assay. CONCLUSIONS: CD34(-) cells provide only a minor contribution to hematopoietic engraftment in this in vivo model system, as compared to CD34(+) cells from the same samples of noncultured human cells. Hematopoiesis derived from actual CD34(-) cells is difficult to distinguish from that due to CD34(+) cells potentially contaminating the preparations. PMID- 11438235 TI - Preface. PMID- 11438236 TI - Interactions of some local anesthetics and alcohols with membranes. AB - A review of the results obtained by our group in the last decade regarding the interactions of procaine, lidocaine, dibucaine and tetracaine with membranes is presented in the context of the literature data. The action upon membranes, in first approximation monomolecular film of stearic acid spread at the air/water interface used as a membrane model, the modification of biomembrane structure and function using diffraction methods, lipid phase transition, fluidity of lipids and proteins, membrane expansion and platelet aggregation were studied. The thermodynamic knowledge of membrane-alcohol interactions improved by using highly sensitive calorimetric techniques are briefly reported. One of the main conclusions is that the physical state of a monolayer model membrane was the result of competitive interactions between film-film and film-substrate interactions. It was taken into account that local anesthetics, such as lidocaine, carbisocaine, mesocaine, showed changes in the bilayer structure, reflected in macroscopic mechanical properties. This restructuring of the lipid bilayer has a significant influence on the operation of functional subunits, e.g. ionic channels formed by gramicidin. The results support the concept of non specific interactions of local anesthetics with lipid bilayers. The theoretical modeling of the interactions of local anesthetics is closely compared with experimental data. Our new theory of relaxation for these interactions is using a non-archimedean formalism based on a process resulting from superpositions of different component processes which take place at different scales of time. PMID- 11438237 TI - Volumetric study on the protein-anesthetic binding. AB - Thermodynamic equations describing the volume behavior of protein-ligand mixtures in water were derived. In order to estimate the volume and binding parameters, the equations were combined with a Langmuir-type binding isotherm. Densities of aqueous solutions of mixtures of bovine serum albumin (BSA) and octanol (C8OH) were measured as a function of total BSA molality, m(M)(T), at constant total C8OH molalities, m(X)(T). The data were analyzed by the equations. The partial molar volumes at infinite dilution of BSA and C8OH, V(M)(T,0) and V(X)(T,0), respectively, were estimated. It was seen that V(M)(T,0) decreases by the addition of C8OH to the solution and that V(X)(T,0) decreases gradually with increasing m(M)(T) and approaches asymptotically to a certain value at high m(M)(T). From the concentration dependence of V(M)(T,0) and V(X)(T,0), the values of the association constant K=392 kg mol(-1), the maximum binding number b(max)=1.9, and the volume change DeltaV=-109 cm(3) mol(-1) were obtained for BSA C8OH interaction in water. The negative value of DeltaV indicates that the hydrophobic interaction reduces the protein volume and elevation of pressure promotes BSA-C8OH binding. These results is inconsistent with the pressure reversal of anesthesia. PMID- 11438238 TI - Alkane derivative-bacteriorhodopsin interaction: proton transport and protein structure. AB - The effects of alkane derivatives, 1-alcohols (ROH), aliphatic amine hydrochlorides (RNH(2).HCl) and sodium aliphatic carboxylates (ROONa), on the proton pumping activity of bacteriorhodopsin (bR) in a purple membrane have been examined. Photocurrents in bR in the purple membrane adsorbed onto polyester thin film were recorded before and after exposure to these test substances. The peak photocurrent in bR was reversibly suppressed by each substance. From the dose response curve, the concentrations required to reduce the peak capacitive current by 50% were determined for each homolog and then the standard free energies per CH(2) for the adsorption of the alkane derivatives to the site of action were estimated: -3.13 kJ mol(-1) for ROH, -3.05 kJ mol(-1) for RNH(3)(+), and -2.95 kJ mol(-1) for ROO(-). The proton pumping activity of bR was mainly suppressed by the hydrophobic interaction with the additive. The relative potencies of the functional groups of the alkane derivatives were almost comparable between 1 octanol (C(8)OH) and octylamine hydrochloride (C(8)NH(3)(+)) and about 10 times less effective for sodium octanoate (C(8)OO(-)) than for others. The addition of C(8)OH or C(8)OO(-) changed the absorption spectra of bR with a maximum at 560 nm to the spectra of the intermediate state with a maximum at 480 nm, while the C(8)NH(3)(+) decreased the intensity of the 560 nm band only with no blue-shift by the 480 nm band. We conclude that the action of the alkane derivatives is nonspecific and directed to all organized purple membrane structures and that the binding sites of the ROH and ROO(-) are different from that of RNH(3)(+). PMID- 11438239 TI - Sensitivity to diethylether anesthesia of fruit flies primarily depends on the genotypes of the sodium channel gene rather than the states of the membranes and the mechanisms might be different from heat-induced paralysis. AB - The para locus of Drosophila melanogaster encodes the alpha subunit of a voltage sensitive sodium channel. Many of the mutants develop paralysis at the high temperature (37 degrees C) and are hypersensitive to diethylether anesthesia. We examined whether the two aspects of the phenotype are mediated by a same mechanism that involves the sodium channel molecule by investigating properties of the three para alleles (para(hd838), para(ts1) and para(ts3)). Larvae of the all para strains showed almost normal sensitivities to diethylether anesthesia while adult flies of them showed hypersensitivities to that in the following manner: para(hd838)<==para(ts1)25) group (mortality rate 28% in women versus 33% in men). This difference was not attributable to mechanism of injury, severity of injury, or pattern of injury. CONCLUSIONS: Severely injured women younger than 50 years of age have a survival advantage when compared with men of equal age and injury severity. Young men have a 27% greater chance of dying than women after trauma. We conclude that gender dimorphism affects the survival of patients after trauma. PMID- 11438263 TI - Limits of peritoneal cytokine measurements during abdominal lavage treatment for intraabdominal sepsis. AB - BACKGROUND: Monitoring of peritoneal cytokine concentrations of tumor necrosis factor (TNF)-alpha was recommended for early detection of severe postoperative complications. In the present study the clinical application of cytokine monitoring was examined in the treatment course of severe peritonitis. METHODS: Nineteen patients with secondary peritonitis were followed up during 75 abdominal lavages. Serum and peritoneal interleukin (IL)-6, IL-8, and IL-10 and TNF-alpha were measured before the surgical intervention, after 1 hour, 3 hours, 6 hours, and 24 hours. Additionally, cardiorespiratory parameters, osmolarity, C-reactive protein, and total leucocyte count were recorded. RESULTS: Serum and peritoneal cytokine concentrations did not correlate to each other as well as to the observed cardiorespiratory parameters. Peritoneal cytokine concentrations were 10 to 1000-fold higher to serum concentrations and showed an intermittent wash out. There were no differences in determined cytokine concentrations between survivors and nonsurvivors. CONCLUSIONS: Once elevated, peritoneal cytokine measurements offer no new diagnostic or prognostic tool in abdominal lavage peritonitis treatment. PMID- 11438264 TI - Comparison of different surgical repairs in the treatment of experimental duodenal injuries. AB - BACKGROUND: In this experimental study, we aimed to investigate the results of different surgical repair methods for delayed reconstruction of severe experimental duodenal defects. METHODS: A large duodenal defect with irregular and tagged margins covering about 50% of the circumference was created in the second part of duodenum of male Wistar rats. The effectiveness of primary repair, jejunal serosal patch, Roux-en-Y duodenojejunostomy, or expanded polytetrafluoroethylene patch repair techniques were investigated on the basis of survival and histologic assessment. RESULTS: No significant survival benefit was observed between jejunal serosal patch, Roux-en-Y duodenojejunostomy, or expanded polytetrafluoroethylene patch repair techniques. But these repair modalities were associated with better survival rates than no-treatment or primary repair techniques. Complete coverage of the expanded polytetrafluoroethylene grafts by neomucosa consisting of columnar epithelium with villus formation was observed in surviving rats about 16 weeks after surgery. CONCLUSIONS: Expanded polytetrafluoroethylene patch can be used in the repair of experimental large duodenal defects, which can not be repaired primarily. PMID- 11438265 TI - Adverse prognostic significance of infraclavicular lymph nodes detected by ultrasonography in patients with locally advanced breast cancer. AB - BACKGROUND: Ultrasonography is increasingly used to evaluate the nodal status of breast cancer patients and specialized positioning permits assessment of the infraclavicular fossa. However, the incidence and significance of infraclavicular (level III) adenopathy detected sonographically in locally advanced breast cancer (LABC) has not been defined. METHODS: The study population consisted of 146 LABC patients registered in a prospective trial of induction chemotherapy between 1991 and 1996. All patients underwent ultrasound imaging before and after chemotherapy. Median follow-up was 32 months. RESULTS: Forty-two of 146 patients (29%) had suspicious infraclavicular adenopathy; all 42 had additional positive axillary lymph nodes by ultrasound. Disease-free and overall survival for the patients with suspicious infraclavicular adenopathy was significantly worse compared with patients without this feature; disease-free survival 50% versus 68% (P = 0.112); overall survival 58% versus 83% (P = 0.026). CONCLUSIONS: Nearly one third of LABC patients will have infraclavicular lymph node involvement by ultrasound imaging; this finding is a significant adverse prognostic feature, and we recommend that infraclavicular nodal evaluation become a routine component of the sonographic workup of breast cancer patients, particularly if lower axillary lymph nodes appear involved. PMID- 11438266 TI - Complications of endoscopy. AB - BACKGROUND: Although most gastrointestinal endoscopic procedures are performed by gastroenterologists, surgeons often assist in the management of patients with complications. This review provides an introduction to the incidence, prevention, and treatment of complications that may occur after upper endoscopy, colonoscopy, percutaneous endoscopic gastrostomy, and endoscopic retrograde cholangiopancreatography. METHODS: Systematic review of the literature. RESULTS: Preprocedural complications include medication effects and adverse effects of bowel preparation. Major procedural complications consist primarily of perforation and hemorrhage. Percutaneous endoscopic gastrostomy tube placement may be complicated by fistula and obstruction. There is also a risk of infectious disease transmission, both to and from the patient. CONCLUSIONS: Endoscopy, like all invasive procedures, carries significant potential risks for the patient. In practiced hands, and with awareness of the problems that may arise, many complications may be avoided and others successfully managed. PMID- 11438267 TI - The reliability of an instrument for identifying and quantifying surgeons' teaching in the operating room. AB - BACKGROUND: The operating room is an important teaching venue where surgical residents develop their operative skills and clinical judgement. The purpose of this study was to establish reliability for an observation instrument designed to quantify surgeons' teaching behaviors in the operating room. METHODS: An instrument was developed to identify operating room teaching behaviors in four categories: informing, questioning, responding, and setting tone. Two trained observers coded videotaped operations. Cronbach's alpha was used to estimate the instrument's internal consistency, and criterion-related reliability was established through an interobserver agreement level (IOA). RESULTS: Results for each of the teaching behavior categories were as follows: informing (IOA = 86%, alpha = +0.978); questioning (IOA = 97%, alpha = +0.966); responding (IOA = 93%, alpha = +0.97); setting tone (IOA = 97%, alpha = +0.882). CONCLUSIONS: This instrument is reliable in identifying and quantifying surgeons' teaching behaviors in the operating room. Identifying teaching behaviors will be valuable to describing and enhancing teaching in the operating room. PMID- 11438268 TI - Development of physical examination skills in a third-year surgical clerkship. AB - BACKGROUND: Previous studies have revealed deficiencies in physical examination (PE) skills of third-year medical students. The purpose of this study was to determine if formal teaching of PE skills and more supervised experiences with clinical faculty would increase the competency of these skills. METHODS: Forty nine third-year medical students were assigned to one of two groups: group A was given formal instruction on PE skills at the beginning of the clerkship, and group B received the same instruction at the midpoint of the clerkship. The two groups received an assessment of the PE skills before, in the middle, and at the end of clerkship, utilizing a physician-developed checklist. RESULTS: Group A had a pretest score of 57.7%. Group B scored 59.8% on the pretest. Independent groups t tests were used to compare the two groups. At the midterm examination, group A increased their score to 71.0%, while group B decreased their score to 55.6%. From beginning to final, both groups had significant improvements in all areas, with group A scoring an overall mean of 75.1% (P = 0.0001), and group B scoring 75.5% (P = 0.0001). CONCLUSION: These findings show the effectiveness of structured learning situations for improving third-year surgery students' PE skills. There is also an indication that usual clerkship activities may not enhance learning of PE skills. PMID- 11438269 TI - Measuring medical students' experience with surgical problems and procedures. AB - BACKGROUND: Measuring medical students' experience on their surgical clerkship rotations to assess the adequacy of the breadth of exposure is essential for producing generalist clinicians. METHODS: A Surgical Clinical Checklist was developed by surveying the surgical faculty for those surgical problems and procedures that every generalist physician should experience. The checklist was then distributed to 48 consecutive third-year medical students for completion during their core clerkship in surgery. RESULTS: Students reported encounters with surgical procedures more frequently than with surgical problems (64.3% of procedures versus 21.9% of problems were encountered by 80% of respondents). Students actively participated as often as they reported passive observation alone. Students assigned to two different teaching sites encountered similar numbers of items at each site although the distribution of individual items was different. CONCLUSIONS: The Surgical Clinical Checklist provides a valuable measurement tool to assess student experiences on their surgical clerkship and can be used to direct future teaching initiatives. PMID- 11438270 TI - Hepatic resection for large hepatocellular carcinoma. AB - BACKGROUND: Long-term survival and prognostic factors after hepatic resection for large hepatocellular carcinoma (HCC) remain to be proved. METHODS: The surgical outcome in 133 consecutive patients with HCC in diameter of > or = 5 cm (large HCC; L group) undergoing hepatic resection was retrospectively clarified and compared with that of 253 patients with HCC in diameter of < 5 cm (small HCC; S group). Postresection prognostic factors were evaluated by univariate and multivariate analysis using Cox's proportional hazards model. RESULTS: The disease-free 3- and 5-year survival rates between L group and S group were 26% versus 42% and 20% versus 25%, respectively (P = 0.0032). The overall 3- and 5 year survival rates between L group and S group were 38% versus 67% and 28% versus 47%, respectively (P < 0.0001). Multivariate analysis revealed that large amount of intraoperative blood transfusion was an independently significant factor of poor disease-free and overall survivals. CONCLUSIONS: Long-term survival in patients with large HCC remains unsatisfactory compared with that in patients with non-large HCC. Restriction of intraoperative blood transfusion may play an important role in the improvement of survival and recurrence in such patients. PMID- 11438271 TI - Modified Seldinger technique for the insertion of standard chest tubes. AB - Closed tube thoracostomy is a standard procedure for the evacuation of air, blood, or other materials from the pleural space. This paper describes a modification of the Seldinger technique that facilitates chest tube insertion. Either a Nelaton or Thieman catheter is threaded into the side drainage hole and out the tip of a standard Argyle-type chest tube. After using the clamp to insert the catheter into the pleural space through a previously dissected tract, the catheter serves as a guide over which the chest tube is inserted. The technique is simple to use, effective, and safe. It employs standard, inexpensive materials to insert chest tubes in such a way as to minimize the potential traumatic complications inherent in other techniques. PMID- 11438272 TI - Prognostic significance of carcinoembryonic antigen levels in peritoneal washes in patients with gastric cancer. AB - BACKGROUND: Peritoneal metastasis is the most frequent cause of death in patients with gastric cancer. Detection of free cancer cells in the peritoneal cavity at the time of surgery, therefore, is considered to be of great value in predicting the peritoneal recurrence and accordingly in the prognosis in patients with gastric cancer. This study examined the clinical significance of intraoperative determination of carcinoembryonic antigen (CEA) levels in peritoneal washes (pCEA) in patients with gastric cancer. METHODS: CEA levels in peritoneal washes were correlated retrospectively with several clinicopathologic factors including clinical outcome in 56 patients with resectable gastric cancer. RESULTS: Among several clinicopathologic factors, the depth of tumor invasion significantly and independently correlated with pCEA levels as revealed by multivariate stepwise logistic regression analysis. A significant difference in overall survival rates was observed between pCEA-positive and pCEA-negative groups: 5-year survival rates were 95.7% in pCEA-negative and 20% in pCEA-positive patients (P <0.0001). Multivariate analysis indicated that pCEA level is a statistically significant independent prognostic factor for the survival of patients with gastric cancer, and is an important factor for predicting peritoneal recurrence. CONCLUSIONS: pCEA could be a potential predictor of a poor prognosis as well as peritoneal recurrence in patients with gastric cancer. We believe that this information could contribute to determining the optimal intraoperative and postoperative therapeutic plan including adjuvant chemotherapy of gastric cancer. PMID- 11438273 TI - Lymphoma presenting as a large ulcerating small bowel lesion. PMID- 11438274 TI - V-Y island flaps for repair of large perianal defects. AB - BACKGROUND: Some perianal diseases such as Paget disease and Bowen disease are extensive and require a wide circumferential excision including the entire anoderm of the anal canal. METHODS: We describe a technique of V-Y island flaps to cover the large perianal defects and the denuded anal canal. It is important to excise the base of the flaps in order to fit them into the anal canal. RESULTS: There were 10 women and 5 men with an average age of 54 years (range 32 to 77). The mean follow-up was 45 months (range 6 to 92). The underlying pathology included various kinds of neoplastic and nonneoplastic diseases. There were no major complications such as flap loss or infection. Most complications were minor, including superficial wound separation, flap hematoma, and anal stricture. Although initially all patients had some degree of incontinence for gas and liquid stool or discharge, none of them had significant fecal incontinence at the time of last follow-up. A diverting ileostomy or colostomy was created in 5 patients. Its role was not clear but it did help in the management of the wounds and minimized the pain. PMID- 11438275 TI - Does patient position during liver surgery influence the risk of venous air embolism? AB - BACKGROUND: It is generally believed that positioning of the patient in a head down tilt (Trendelenberg position) decreases the likelihood of a venous air embolism during liver resection. METHODS: The physiological effect of variation in horizontal attitude on central and hepatic venous pressure was measured in 10 patients during liver surgery. Hemodynamic indices were recorded with the operating table in the horizontal, 20 degrees head-up and 20 degrees head-down positions. RESULTS: There was no demonstrable pressure gradient between the hepatic and central venous levels in any of the positions. The absolute pressures did, however, vary in a predictable way, being highest in the head-down and lowest during head-up tilt. However, on no occasion was a negative intraluminal pressure recorded. CONCLUSION: The effect on venous pressures caused by the change in patient positioning alone during liver surgery does not affect the risk of venous air embolism. PMID- 11438276 TI - Learning preferences, computer attitudes, and test performance with computer aided instruction. AB - BACKGROUND: Learning preference refers to how individuals choose to approach learning situations. Computer-aided instruction (CAI) permits the adaptation of educational content to individual student learning strategies. METHODS: To determine if learning preference and computer attitude influence the acquisition of knowledge using CAI materials, a prototype CAI program was developed that incorporated differing learning exercises. Students (n = 180) completed Rezler's Learning Preference Inventory (LPI) and a computer attitude survey (CAS). The LPI uses three sets of paired scales to characterize learning preference and choice of learning situation. The CAS assesses student attitudes toward computers in general (CAS-G), as well as the educational use of computers (CAS-E). After finishing the program students completed a program attitude survey (CAS-P). Immediate comprehension was assessed by pretests and posttests incorporated into the program. Retention was assessed by a repeat of the posttest 4 to 6 weeks after initial program review. RESULTS: Scores (mean +/- SEM) on the pretest, posttest, and late posttest were 38.1% +/- 1.35%, 70.9% +/- 1.24%, and 62.5% +/- 1.44%, respectively. There was no correlation between students' learning preferences or computer attitude and test performance. CONCLUSIONS: The data indicate that CAI provides a means of delivering educational content that results in an increase in knowledge that is not correlated with computer attitudes or learning preferences. PMID- 11438277 TI - Long-term results of duodenectomy with highly selective vagotomy in the treatment of complicated duodenal ulcers. AB - BACKGROUND: Highly selective vagotomy and complete circular or partial duodenectomy have been applied to complicated duodenal ulcer for many years. These procedures seem to provide better clinical results than truncal vagotomy and antrectomy. METHODS: A retrospective analysis was conducted of 120 patients with complicated duodenal ulcer who underwent surgical treatment between 1986 and 1999. Patients with obstruction were treated with either circular complete (17) or partial duodenectomy (3) combined with highly selective vagotomy or truncal vagotomy and antrectomy (37). Those with perforation were treated primarily with highly selective vagotomy and partial duodenectomy, highly selective vagotomy alone, or truncal vagotomy and pyloroplasty. Every patient was followed up either by a clinic visit (75%) or questionnaire to determine the presence of ulcer pain, dumping, diarrhea, vomiting, weight loss, and Visick grade. RESULTS: Long-term follow-up of patients treated with duodenectomy and highly selective vagotomy for obstruction showed that 94% had sustained weight gain whereas more than half of those treated with truncal vagotomy and antrectomy had weight loss. In patients with perforation, duodenectomy and highly selective vagotomy offered no advantage over highly selective vagotomy alone. CONCLUSIONS: Highly selective vagotomy and complete circular or partial duodenectomy provide fewer sequelae and better weight gain long term than truncal vagotomy and antrectomy for patients with obstructing duodenal ulcers. PMID- 11438278 TI - Laparoscopic repair of superior mesenteric artery syndrome. AB - BACKGROUND: Superior mesenteric artery syndrome is duodenal obstruction by the superior mesenteric artery. It is caused by decreasing the angle between the aorta and superior mesenteric artery causing compression of the third part of the duodenum and usually occurs after a period of weight loss. METHODS: Between September 1999 and April 2000, 2 patients with superior mesenteric artery syndrome were treated laparoscopically. The laparoscope was placed in the umbilicus; the surgeon operated through two trochars on the left side of the abdomen, and an assistant retracted through one trochar on the right side of the abdomen. The dilated duodenum was seen below the transverse mesocolon and to the right of the superior mesenteric artery. A proximal loop of jejunum was anastamosed to the duodenum using the endoscopic gastrointestinal anastomotic (GIA) stapler. RESULTS: Average operating time was 113 minutes and average hospital length of stay was 3 days. There were no complications and both patients were pleased with their results. CONCLUSIONS: Laparoscopic duodenojejunal bypass is feasible with laparoscopic techniques. The operating time is acceptable and the postoperative length of stay is short. PMID- 11438279 TI - Femoral pseudoaneurysm thrombinjection. AB - BACKGROUND: The risk of pseudoaneurysm (PSA) increases with the number of catheterizations performed for cardiovascular diagnosis and therapy. Thrombin injection, or "thrombinjection," is an alternative to ultrasound-guided compression or surgical repair. Thrombinjection is effective and economical, but the specter of systemic arterial thrombosis hinders its wide application. We report cautionary steps taken prior to injection and lessons learned during our first 20 cases. METHODS AND RESULTS: Thrombinjection was performed under ultrasound (US) guidance in 12 women and 8 men with PSA after femoral catheterization. PSA varied in size from 17 to 39 mm with neck openings from 1.5 to 3.9 mm. Under local anesthesia and US longitudinal view, a 20-gauge US needle tip was placed in the PSA away from the neck. With 12 cases under 600 units, 100 to 2,300 units of thrombin (1,000 U/mL) were injected slowly. Slow injection, positioning of the ball of thrombus formed at the needle tip, probe compression, and combined or interchangeable use of US color flow and B-mode were the essential techniques utilized. All PSA thrombosed successfully on the first treatment. One patient had fever and another returned for compression treatment of a second, not recurrent, PSA. CONCLUSIONS: Successful thrombinjection was accomplished by focusing on common femoral artery PSA with small neck openings, avoiding arteriovenous fistulas, and using standard thrombin concentration, US needle, state of the art ultrasonography, slow injection, thrombus ball positioning, and adjunctive probe compression. PMID- 11438280 TI - Analysis of phospholipid molecular species by liquid chromatography--atmospheric pressure chemical ionisation mass spectrometry of diacylglycerol nicotinates. AB - A method using liquid chromatography - atmospheric pressure chemical ionisation mass spectrometry was evaluated for determining the molecular species composition of phospholipids (phosphatidylcholines from soybean, egg yolk and bovine liver) after conversion to diacylglycerol nicotinate derivatives. The structures could be deduced from pseudo-molecular ions ([MH-123](+)) and three pairs of monoacyl containing fragment ions. All molecular species in mixed peaks were readily identified and many minor components, earlier not encountered in the samples under investigation, were identified. Acyl chain regioisomers were readily distinguished by the ratio of the [MH-RCHCO](+) ions. Molecular species differing only in the position of the double bonds in one polyunsaturated acyl chain were separated on the basis of retention times. A half quantitative estimation of the molecular species composition of complex samples was achieved by a combination of UV detection and, for mixed peaks, the areas of [MH-123](+) ions. PMID- 11438281 TI - Photophysics of thionine dye in aqueous and liposome media in presence of different reducing agents. AB - The photoelectrochemical and spectral (both absorption and fluorescence) studies of thionine, a cationic phenothiazine dye, have been carried out in aqueous and phosphatidylcholine liposome media in the presence of different reducing agents, such as I(-), Br(-), Cl(-) and Fe(2+). The results show that the photovoltage generation from photoelectrochemical studies and Stern-Volmer quenching constant studied by fluorescence quenching support the photoinduced electron transfer from the reducing agent to the singlet excited thionine dye. Moreover, a good correlation between photovoltages/Stern-Volmer quenching constants vs. reduction potentials of the reducing agents also confirms the above electron transfer in the photoexcited state. PMID- 11438282 TI - Novel azido fatty acid ester derivatives from conjugated C(18) enynoate. AB - Methyl octadec-11Z-en-9-ynoate (1) was epoxidized to give methyl 11,12-Z-epoxy octadec-9-ynoate (2, 81%). Acid catalyzed ring opening of the epoxy ring of compound 2 gave methyl 11,12-dihydroxy-octadec-9-ynoate (3, 80%). The latter was treated with mesyl chloride to yield methyl 11,12-dimesyloxy-octadec-9-ynoate (4, 76%). Reaction of compound 4 with sodium azide furnished methyl 11-azido-12 mesyloxy-octadec-9-ynoate (5a, 49%) and methyl 11-azido-octadec-11E-en-9-ynoate (5b, 24%). Compound 2 was semi-hydrogenated over Lindlar catalyst to give methyl 11,12-Z-epoxy-octadec-9Z-enoate (6, 90%). This allylic epoxy fatty ester (6) was reacted with sodium azide to give a mixture of methyl 11-azido-12-hydroxy-octadec 9Z-enoate (7a) and methyl 9-azido-12-hydroxy-octadec-9E-enoate (7b), which could not be separated into individual components by silica chromatography. Chromic acid oxidation of the mixture of compounds 7a and 7b furnished methyl 9-azido-12 oxo-octadec-10E-enoate (8, 42% based on amount of compound 6 used) and an intractable mixture of polar compounds. The various products were characterized by NMR spectroscopic and mass spectral analyses. PMID- 11438283 TI - The effect of arbutin on membrane integrity during drying is mediated by stabilization of the lamellar phase in the presence of nonbilayer-forming lipids. AB - Arbutin (4-hydroxyphenyl-beta-glucopyranoside) is a solute accumulated to high concentrations in drought and frost resistant plants. Arbutin can inhibit membrane lysis, both free radical-mediated and enzymatic in nature, and it has been suggested that arbutin might contribute to membrane stabilization in these plants. However, we found that arbutin destabilized phosphatidylcholine vesicles during drying and rehydration, which appears to be inconsistent with the proposed protective function of arbutin for membranes. We also found, however, that arbutin stabilizes membranes containing nonbilayer-forming lipids during freezing. We now report that, in liposomes containing the nonbilayer-forming lipids monogalactosyldiacylglycerol (MGDG) or phosphatidylethanolamine (PE), arbutin served a protective function during drying, as measured by retention of carboxyfluorescein (CF) and extent of vesicle fusion. In hydrated samples containing these lipids, arbutin stabilized the lamellar liquid crystalline phase. Therefore, the interaction between arbutin and lipid membranes and the resulting effects on membrane stability depend, in a complex manner, on the lipid composition of the membrane. PMID- 11438284 TI - DNA condensation by an oxidizable cationic detergent. Interactions with lipid vesicles. AB - Cationic amphiphile-mediated delivery of plasmid DNA is the non-viral gene transfer method most often used. In the present work, we considered a new cysteine-detergent, ornithinyl-cysteinyl-tetradecylamide (C(14)-CO), able to convert itself, via oxidative dimerization, into a cationic cystine-lipid. By using fluorescence techniques, we first characterized the structure of complexes of plasmid DNA with C(14)-CO molecules either kept as monomers, or oxidized into dimers. Both forms are able to condense DNA, with the formation of hydrophobic micelle-like domains along the DNA chain. Domains with a larger molecular order were obtained with dimeric C(14)-CO/DNA complexes. In a second step, the interactions of these complexes with lipid vesicles considered as membrane models were investigated. In the presence of vesicles, we observed a decondensation of the DNA involved in complexes obtained with C(14)-CO monomers. With anionic vesicles, the DNA is released into the bulk solution, while with neutral vesicles, it remains bound to the vesicles via electrostatic interactions with inserted C(14)-CO molecules. In sharp contrast, the complexes with C(14)-CO dimers are unaffected by the addition of either neutral or anionic vesicles and show no interaction with them. These results may partly explain the low transfection efficiency of these complexes at the +/-charge ratios used in this study. PMID- 11438285 TI - Surface behaviour of bile salts and tetrahydrolipstatin at air/water and oil/water interfaces. AB - The surface behaviour of two bile salts, sodium deoxycholate (NaDC) and sodium taurodeoxycholate (NaTDC), as well as that of tetrahydrolipstatin (THL), a potent gastrointestinal lipase inhibitor, was studied at air/water and oil/water interfaces, using interfacial tensiometry methods. The surface behaviour of NaDC and NaTDC was comparable at both oil/water and air/water interfaces. A fairly compact interfacial monolayer of bile salts is formed well below the critical micellar concentration (CMC) and can help to explain the well-known effects of bile salts on the kinetic behaviour of pancreatic lipases. Using the Wilhelmy plate technique, the surface pressure-molecular area curves recorded with THL at the air/water interface showed a collapse point at a surface pressure of 24.5 mN.m(-1), corresponding to a molecular area of 70 A(2). Surprisingly, using the oil drop method, a limiting molecular area of 160 A(2) was found to exist at the oil/water interface. On the basis of the above data, space-filling models were proposed for bile salts and THL at air/water and oil/water interfaces. PMID- 11438286 TI - Superoxide dismutase inhibits lipid peroxidation in micelles. AB - Superoxide dismutase (SOD) taken in minor concentrations (a few U/ml) displays a pronounced inhibiting effect on the chain oxidation of methyl linoleate and methyl linolenate (but not methyl oleate) induced by 2,2'-azobis(2-amidinopropan) dihydrochloride (AAPH) in micellar solutions of sodium dodecyl sulfate and Triton X-100 in phosphate buffer, pH 7.40, at 37.0 degrees C. The inhibition is evidently caused by purging the system from O(2)*(-). The latter suggests the formation of O(2)*(-) (HO(2)* in the course of peroxidation, most likely, via beta-decay of lipid peroxy radical (LO(2)*. Thermodynamic estimations verify a rather high probability of beta-decay of LO(2)* produced from polyunsaturated fatty acids by contrast to that produced from saturated and monoenic fatty acids. It is speculated that O(2)*(-) (HO(2)*, being an amphiphilic, reactive and highly mobile species, participates in intermicellar (interliposomal) transfer of free valence during lipid peroxidation in microheterogeneous systems. PMID- 11438288 TI - European Communities Confederation of Clinical Chemistry Working Group on Accreditation: past, present and future. AB - During the past 10 years, many activities have taken place in the field of quality systems and accreditation in medical laboratories. Each country in Europe has a slightly different approach. The Working Group on Accreditation of the European Communities Confederation of Clinical Chemistry (EC4) tries to support harmonisation of these efforts. For this purpose, they edited the Essential Criteria for quality systems of medical laboratories and supported the forthcoming International ISO standard "Quality management for the medical laboratory". At this moment, a Model Quality Manual is nearly ready for publication. The next items are setting up criteria for auditing the quality system and criteria for the accreditation process. PMID- 11438289 TI - Accreditation and postgraduate training in European countries: an FESCC survey. Federation of European Societies of Clinical Chemistry. AB - The mission of the Federation of European Societies of Clinical Chemistry is to support and promote clinical chemistry and laboratory medicine in Europe, to aid communication between the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) and National Scientific Societies, to develop education and quality in the discipline, and to encourage young scientists to take an active role in these activities. One recent initiative of the Federation was a survey on accreditation of medical laboratories and training in laboratory medicine in Europe. Among European countries, three promote accreditation according to EN 45001, several countries apply systems based on professional and nationally defined standards (e.g. CPA in the UK) and other countries are moving to define accreditation standards at a national level. Data on vocational training demonstrate that this is based on a postgraduate education with duration ranging from 6 months to 11 years; in most countries the average length of university education is 5 years and that of postgraduate training is 4 years. A great difference, moreover, exists regarding the polyvalent versus monovalent training. Taken together, these data indicate that a great effort should be made by the Federation for promoting harmonization and coordination in Europe. PMID- 11438290 TI - ISO and CEN documents on quality in medical laboratories. AB - The forthcoming international standard ISO 15189 "Quality management in the medical laboratory" is a document of great importance for the development of quality systems and accreditation for medical/clinical laboratories. For the first time, there will be an internationally recognized standard designed specifically for the accreditation of medical laboratories. The document takes into account the special requirements imposed by the medical environment and by the essential contribution of the medical laboratory service to patient care. It recognizes that medical laboratories must provide not only testing of patient samples, but also advisory, interpretative and educational services. A further document, still in draft form (ISO/DIS 15190), deals with safety management for medical laboratories. ISO 15189 (and probably 15190 also) are expected be adopted by CEN as a European Standard (EN). PMID- 11438291 TI - The EC4 quality manual model. AB - One of the priorities of the European Confederation of Clinical Chemistry (EC4) is the harmonisation of the clinical laboratory profession in Europe. One of the first steps is to try to harmonise the quality systems, that is, the clinical laboratory organisational structure, responsibilities, procedures, processes and resources involved in quality management. The "EC4 Essential Criteria" were published by the Working Group on Harmonisation of Quality Systems in order to facilitate the development or the update of a quality system in a clinical laboratory, and to encourage international bodies to produce specific Standards for the clinical laboratory. Furthermore, the EC4 Working Group has produced a Quality Manual Model, which includes a sample of quality policy documents and some operational directions for an imaginary laboratory. This Quality Manual Model was prepared following the "EC4 Essential Criteria." Its purpose is that any quality system developed following the Manual could be accredited or certified against any Standard. The EC4 Quality Manual Model will be available, free of charge, to clinical laboratory professionals. PMID- 11438292 TI - Standards for the medical laboratory--harmonization and subsidiarity. AB - An International Standard, ISO 15189, specifically for 'Quality management in the medical laboratory' for use by accrediting 'bodies that recognize the competence of medical laboratories,' is expected to be published shortly. The origins, content and limitations of the new standard are discussed and the diversity of current arrangements for accreditation is reviewed. A new International Standard is an important step towards harmonization of laboratory practice but an accreditation system is more than its standards and a harmonized approach to the treatment of noncompliances found at inspection is important. Experience gained in writing national standards can improve the approach to the drafting and improvement of International Standards. Recognition of the principle of subsidiarity aids rather than hinders progress to harmonization and empowers the 'fourth element' (the laboratories to be accredited) to be a part of the accreditation process. PMID- 11438293 TI - Role of inspectors in external review mechanisms: criteria for selection, training and appraisal. AB - There is a wide consensus that an external review mechanism, both in the form of a peer review, accreditation and certification according to the ISO 9000 series, is more than its standards. The survey process, the role of inspectors and standard interpretation contribute to the essence of the programme itself. Above all, the criteria used for the selection, training and appraisal of inspectors are of paramount importance. While the ISO norms do not require certification bodies to employ "peer reviewers" for the healthcare sector, experience in this sector is the main criterion for recruiting inspectors in accreditation and peer review programmes. However, the ISO/IEC Guide 58, for the setting up and operation of a laboratory accreditation body, specifies that inspectors should have appropriate technical knowledge of the specific calibrations, tests or types of calibration or tests for which accreditation is sought. Training, updating and assessment of inspectors are clearly defined under ISO, but are also systematic under accreditation programmes. Part-time inspectors who are professionals currently practising in a healthcare facility and are in touch with the day-to day work reality are preferred for accreditation programmes which have self regulation, education and quality improvement as their main concerns, while full time and external inspectors are used in external review mechanisms with registration and certification as their main concerns. As well as harmonising the standards for accreditation, it is important to obtain consensus on the criteria to use for the selection, training and assessment of inspectors in order to ensure that different national or international programmes gain mutual recognition. PMID- 11438294 TI - Accreditation of medical laboratories in The Netherlands. AB - Among the medical laboratory professionals, a growing interest exists in the systematic application of quality assurance to their own practice, and consequently in The Netherlands, there is an increasing request for an official accreditation of the quality management system and the competence of the professionals through Coordinatie Commissie voor Kwaliteitsbewaking in Laboratoria in de gezondheidszorg (CCKLtest), the Dutch Accreditation Board for Medical Laboratories. This article gives an overview of the current situation in The Netherlands; regarding the standards, the rules for accreditation, the training and selection assessors and what meanwhile is achieved. Since 1994, 60 clinical laboratories have been recognized by CCKLtest and another 40 laboratories have requested accreditation and are waiting for the inspection in 2001. Thus, 25% of all clinical laboratories in The Netherlands will be inspected within the current year. PMID- 11438295 TI - Accreditation of medical laboratories. Some reflections from the Nordic Horizon. AB - Accreditation has been a successful approach to improved quality management in laboratories and applied to a wide variety of medical laboratories. After the initial enthusiastic phase, in which quality improvement is significant, the endurance is threatened by an increasing bureaucracy of the process. A productive balance between the accreditation bodies and the profession needs to be sought. New standards have been suggested by ISO and the profession needs to take the initiative to assure that a global standard specially adopted to the requirements of medical laboratories becomes recognised and used. The new concept of uncertainty may become an important tool to further improve and describe the performance of laboratories by allowing inclusion of pre- and post-analytical sources of uncertainty as well as bias. PMID- 11438296 TI - IVD industry role for quality and accreditation in medical laboratories. AB - Manufacturers of in vitro diagnostic (IVD) medical devices and laboratory management have become integral partners in building and improving the quality of laboratory services. There is an increasing awareness that quality is inherent in the design of any reagent or analytical system. In vitro diagnostic medical devices should provide patients, users and third parties with a high level of health protection. Therefore, both manufacturers and users must work in partnership for continual improvement. For manufacturers, standards such as ISO 9000 already exist to guide applications of quality practices. In the field of laboratory medicine, the availability of a specific, universal standard (ISO/DIS 15189) for quality management in medical laboratories will represent a great opportunity for harmonising medical laboratories at an international level. In addition, accreditation of medical laboratories according to the proposed ISO 15189 standard can help develop the relationships between laboratories, and the biological follow-up of travelling patients. Manufacturers are able to help laboratory management to reach a high level of quality, not only by providing high value products, but also on the basis of their own experience of ISO 9000 certification. PMID- 11438297 TI - Role of external quality assurance schemes in assessing and improving quality in medical laboratories. AB - Medical laboratories have a long tradition of external quality assessment. Starting from pure quality control of laboratory performances, most external quality schemes in Europe evolved into a powerful tool for improving quality of clinical outcome of results. External quality assurance in medical laboratories not only includes laboratory performance evaluation, but also evaluation of method performance, post-market vigilance, training and help. In the future, the quality of programmes will further be improved by accreditation of schemes international collaboration and by using IT applications. PMID- 11438298 TI - Accreditation of external quality assessment schemes in the United Kingdom. AB - There are some 130 pathology external quality assessment schemes available in the United Kingdom. Most are provided from within the National Health Service, but a few from the private sector. At the invitation of the Department of Health, Clinical Pathology Accreditation (UK) Ltd. has developed an accreditation system for these schemes. Accreditation is based on scheme organisers declaring compliance with 62 standards and an on-site inspection. As of the end of 2000, 105 schemes have been accredited. Most of the schemes in the disciplines of Clinical Biochemistry, Haematology, Immunology and Microbiology have now been accredited, but progress has been slower in non-numerical schemes in the disciplines of Cellular Pathology and Genetics. PMID- 11438299 TI - External Quality Assessment Schemes: need for recognised requirements. AB - Programs for Accreditation of clinical laboratories consider participation in External Quality Assessment Schemes (EQAS) a key element in the evaluation of testing procedures and improving them. One of the main functions of EQAS is to assess whether laboratories perform tests competently. It is therefore of utmost importance for laboratories to participate in EQAS that are in line with formally recognised requirements. Specific proposals have been made on how to design and execute EQAS by International Working Groups, but there seems to be no consensus on the best strategies to use and quality specifications to set out. The Clinical Pathology Accreditation (CPA) Program for EQA Scheme Accreditation (CPA-EQA) is the only program in Europe to provide a formal recognition of the quality of EQAS activities. The present paper reports on the experience of the Centre of Biomedical Research which is following an accreditation process for their own schemes in line with the CPA-EQA program and a proposal to set requirements that Italian schemes must follow to be recognised as valid and effective. PMID- 11438300 TI - Comparative actions of insulin sensitizers on ion channels in vascular smooth muscle. AB - Thiazolidinedione and isoxazolidinedione insulin sensitizers activate peroxisome proliferator-activated receptor gamma (PPAR gamma). Some thiazolidinediones modify ion channels in smooth muscles; however, the mechanism by which their actions occur has not been clarified. We, thus, examined the effects of three thiazolidinediones (troglitazone, pioglitazone, and rosiglitazone) and isoxazolidinedione (JTT-501), as well as an intrinsic ligand for PPAR gamma, 15 deoxy-Delta(12,14) prostaglandin J(2) (prostaglandin J(2)), on voltage-operated Ca(2+) currents (I(Ca)), voltage-dependent K(+) currents (I(Kv)), and Ca(2+) activated K(+) currents (I(Kca)), to clarify whether a thiazolidinedione structure or PPAR gamma activation is related to their actions on ion channels. The whole-cell patch clamp method was used to record currents in smooth muscle cells from guinea-pig mesenteric arteries. Thiazolidinediones inhibited I(Ca) in a dose-dependent manner (troglitazone>pioglitazone=rosiglitazone). Troglitazone (> or =1 microM) and rosiglitazone (100 microM), but not pioglitazone, inhibited I(Kv). Rosiglitazone (> or =10 microM) enhanced, troglitazone (> or =1 microM) inhibited, and pioglitazone did not affect I(Kca). A high concentration of JTT 501 (100 microM) inhibited I(Ca), I(Kv), and I(Kca) to a similar extent. Prostaglandin J(2) enhanced I(Kca), but affected neither I(Ca) nor I(Kv). In summary, the three thiazolidinediones and isoxazolidinedione act differently on Ca(2+) and K(+) channels in vascular smooth muscle. The action of thiazolidinediones on I(Ca) could be attributed to specific regions of the molecules and not to activation of PPAR gamma. Involvement of PPAR gamma activation in the stimulation of I(Kca) is possible but should be tested further. PMID- 11438301 TI - Effect of nitric oxide on apoptotic activity in the rat gastrointestinal tract. AB - The effect of nitric oxide (NO) on apoptosis in the gastrointestinal mucosa was investigated. Experiments involved long-term exposure of rat gastric mucosal cells in vitro to exogenous NO delivered from the NO, donor S-nitroso-N-acetyl penicillamine, and the effect of intravenous administration of lipopolysaccharide in vivo, in the presence and absence of the selective inhibitor of inducible NO synthase N-(3-(aminomethyl)benzyl) acetamidine (1400 W). S-nitroso-N-acetyl penicillamine produced a dose-related inhibition of caspase 3-like activity and DNA fragmentation in isolated gastric mucosal cells. Caspase 3-like activity and DNA fragmentation in gastric, ileal and colonic mucosa were increased both 5 and 24 h after injection of lipopolysaccharide (3 mg/kg, i.v.) to rats in vivo. Administration of 1400 W (5 mg/kg, i.v.) immediately after lipopolysaccharide enhanced caspase 3-like activity and DNA fragmentation above that found with lipopolysaccharide alone. In conclusion, data obtained both in vitro and in vivo suggest that NO exerts an anti-apoptotic effect on rat gastrointestinal mucosal cells. PMID- 11438302 TI - Uncompetitive NMDA receptor antagonists potentiate morphine antinociception recorded from the tail but not from the hind paw in rats. AB - We investigated the effects of pretreatment with low-affinity, uncompetitive NMDA receptor antagonists on morphine-induced antinociception in rats using the same intensity of thermal stimulus applied to the tail and the paws. Similar baseline responses to thermal stimuli of the same intensity were recorded from tails and hind paws. However, morphine produced equal antinociception from the tail and hind paw when used at doses of 2.5 and 6 mg/kg, respectively. These doses were used in further experiments. Thirty minutes before morphine, rats were administered the NMDA receptor antagonists dextromethorphan (2.5--30 mg/kg), memantine (2.5--15 mg/kg) and MRZ 2/579 (1-amino-1,3,3,5,5-pentamethyl cyclohexane HCl) (1.25--10 mg/kg). All three compounds significantly and dose dependently potentiated morphine-induced antinociception recorded from the tail. However, none of these NMDA receptor antagonists affected morphine antinociception recorded from the paw. These findings suggest that low-affinity NMDA receptor antagonists modulate differently morphine antinociceptive activity recorded from the tail and hind paws. PMID- 11438303 TI - Neuropharmacological profiles of a novel atypical antipsychotic, NRA0562, in rats. AB - Neuropharmacological profiles of 5-(2-[4-(6-fluoro-1H-indole-3-yl) piperidine-1 yl] ethyl)-4-(4-fluorophenyl) thiazole-2-carboxylic acid amide (NRA0562) in rats were examined using electrophysiological and immunohistochemical methods. The firing rates of the substantia nigra pars compacta (A9) and the ventral tegmental area (A10) dopamine neurons were inhibited by methamphetamine (1 mg/kg, i.v.). NRA0562 dose-dependently reversed the inhibitory effects of methamphetamine on A9 and on A10 dopamine neurons. NRA0562 was more potent to reverse the inhibitory effects of methamphetamine on A10 (ED(50)=0.3 mg/kg) than on A9 (ED(50)=0.9 mg/kg) dopamine neurons. NRA0562 produced significant increases in Fos-like immunoreactivity in both the nucleus accumbens and the dorsolateral striatum. The difference between the number of Fos-like immunoreactivity produced by NRA0562 in the nucleus accumbens vs. dorsolateral striatum, referred to as the atypical index, was positive. Similar results could be observed with risperidone, an atypical antipsychotic. These results suggest that NRA0562 may have the atypical antipsychotic activities seen with risperidone, but without the liability of motor side effects typical of classical antipsychotics. PMID- 11438304 TI - Intra-amygdala spermidine administration improves inhibitory avoidance performance in rats. AB - In the present study, we investigated the effect of immediate post-training bilateral infusion of spermidine, a polyamine agonist, into the amygdala on inhibitory avoidance learning of rats. Bilateral microinjection of spermidine (0.02--20 nmol) caused an increase in test step-down latencies at high concentrations. Administration of arcaine (0.002--0.2 nmol), an antagonist of the NMDA receptor polyamine binding site, decreased test step-down latencies. On the other hand, co-administration of arcaine and spermidine completely reversed the spermidine-induced increase of test step-down latencies. These results provide evidence that polyamines may be involved in learning and memory modulation in the amygdala. PMID- 11438305 TI - Role of the NMDA receptor subunit in the expression of the discriminative stimulus effect induced by ketamine. AB - Ketamine, which is a non-competitive NMDA receptor antagonist, has been used as a dissociative anesthetic agent. However, chronic use of ketamine produces psychotomimetic effects, such as nightmares, hallucination and delusion. Therefore, the present study was designed to ascertain the role of the NMDA receptor and sigma receptor in the discriminative stimulus effect induced by ketamine. Fischer 344 rats were trained to discriminate between ketamine (5 mg/kg, i.p.) and saline under a fixed-ratio 10 food-reinforced procedure. Non competitive antagonists for both NR2A- and NR2B-containing NMDA receptors, such as phencyclidine (0.1--1 mg/kg, i.p.) and dizocilpine (3--30 microg/kg, i.p.), and the NR2A-containing NMDA receptor-preferred antagonist dextromethorphan (3- 56 mg/kg, i.p.) fully substituted for the ketamine cue in a dose-dependent manner. By contrast, the NR2B-containing NMDA receptor antagonist ifenprodil (5- 20 mg/kg, i.p.) exhibited no generalization. Additionally, the competitive NMDA antagonist 3-[(+/-)-2-carboxypiperazine-4-yl] propyl-1-phosphonic acid ((+/-) CPP; 0.3--5.6 mg/kg, i.p.) and a sigma receptor ligand DTG (0.3--3 mg/kg, s.c.) displayed no generalization to the ketamine cue. These results suggest that NR1/NR2A subunit containing NMDA antagonism may be critical for the production of the ketamine cue. PMID- 11438306 TI - Factors involved in the time course of response to acetylcholine in mesenteric arteries from spontaneously hypertensive rats. AB - The time course of the response to prolonged application of acetylcholine in mesenteric arteries from stroke-prone spontaneously hypertensive rats (SHRSP) and Wistar Kyoto rats (WKY) was compared. Only a relaxing response, which was blocked by N(omega)-nitro-L-arginine (L-NOARG), was observed after the prolonged application of a low concentration of acetylcholine (10(-8) M) in both preparations; the response was impaired in SHRSP preparations. Prolonged application of a high concentration of acetylcholine (10(-5) M) induced a second contractile response after a first relaxing response in SHRSP preparations under basal conditions and in WKY preparations in the presence of L-NOARG. This contractile response was attenuated by indomethacin. In the presence of a combination of apamin and charybdotoxin, the relaxing response to the high concentration of acetylcholine was reduced and a contractile response, which was abolished by indomethacin, appeared. In the presence of all of these blockers, a contractile response, which was blocked by cyclo(D-alpha-aspartyl-L-propyl-D valyl-L-leucyl-D-tryptophyl) (BQ-123), was observed in preparations from WKY but not in preparations from SHRSP. Results indicate that prolonged application of acetylcholine in rat mesenteric arteries induces the release of endothelium derived relaxing, contracting, hyperpolarizing factors and endothelin-1, and that the mode of action differs between preparations from WKY and SHRSP. PMID- 11438307 TI - Inhibition of prostanoid-mediated contraction to endothelin-1 after hypoxia in rat aorta. AB - The role of the thromboxane A(2)/prostaglandin H(2) receptor in endothelin-1 contraction was investigated in aortic rings from rats exposed to normoxia (21% O(2)) or hypoxia (10% O(2)) for 12 h. Indomethacin (10 microM) and SQ 29,548 (0.1 microM, thromboxane A(2)/prostaglandin H(2) receptor antagonist) reduced maximum tension and increased EC(50) in endothelium-intact and -denuded rings from normoxic animals. Neither inhibitor had any effect on rings from hypoxic rats. Thromboxane A(2) and/or prostaglandin H(2) contribute to the response to endothelin-1 in aortas from normoxic rats but not from rats exposed to hypoxia. Loss of prostanoid-enhancement of endothelin-1 contraction contributes to impair vascular reactivity after hypoxia. PMID- 11438308 TI - Involvement of nitric oxide in pollen-induced biphasic nasal blockage in sensitised guinea pigs. AB - We have developed a reproducible allergic rhinitis model showing biphasic nasal blockage on repetitive inhalation challenge with Japanese cedar pollen in sensitised guinea pigs. The role of nitric oxide (NO) in inducing nasal blockage was evaluated with this model. N(omega)-nitro-L-arginine methyl ester (L-NAME), a non-selective NO synthase (NOS) inhibitor, intravenously administered before the challenge, significantly inhibited both early and late nasal blockage by approximately 80% and 50%, respectively. When L-NAME treatment was performed after the challenge, the late response was inhibited by approximately 70%. This inhibition was completely reversed by co-administration of L-arginine. However, aminoguanidine and L-N(6)-(1-iminoethyl)lysine, selective inhibitors of inducible NOS, negligibly influenced the degree of nasal blockage. Meanwhile, the alpha adrenergic agonist, naphazoline, strongly suppressed both early and late nasal blockage. These results indicate that NO, likely produced by constitutive rather than inducible NOS, plays a major role in the occurrence of biphasic nasal blockage, primarily by inducing vasodilatation. PMID- 11438309 TI - The in vitro pharmacological profile of prucalopride, a novel enterokinetic compound. AB - Prucalopride is a novel enterokinetic compound and is the first representative of the benzofuran class. We set out to establish its pharmacological profile in various receptor binding and organ bath experiments. Receptor binding data have demonstrated prucalopride's high affinity to both investigated 5-HT(4) receptor isoforms, with mean pK(i) estimates of 8.60 and 8.10 for the human 5-HT(4a) and 5 HT(4b) receptor, respectively. From the 50 other binding assays investigated in this study only the human D(4) receptor (pK(i) 5.63), the mouse 5-HT(3) receptor (pK(i) 5.41) and the human sigma(1) (pK(i) 5.43) have shown measurable affinity, resulting in at least 290-fold selectivity for the 5-HT(4) receptor. Classical organ bath experiments were done using isolated tissues from the rat, guinea-pig and dog gastrointestinal tract, using various protocols. Prucalopride was a 5 HT(4) receptor agonist in the guinea-pig colon, as it induced contractions (pEC(50)=7.48+/-0.06; insensitive to a 5-HT(2A) or 5-HT(3) receptor antagonist, but inhibited by a 5-HT(4) receptor antagonist) as well as the facilitation of electrical stimulation-induced noncholinergic contractions (blocked by a 5-HT(4) receptor antagonist). Furthermore, it caused relaxation of a rat oesophagus preparation (pEC(50)=7.81+/-0.17), in a 5-HT(4) receptor antagonist sensitive manner. Prucalopride did not cause relevant inhibition of 5-HT(2A), 5-HT(2B), or 5-HT(3), motilin or cholecystokinin (CCK(1)) receptor-mediated contractions, nor nicotinic or muscarinic acetylcholine receptor-mediated contractions, up to 10 microM. It is concluded that prucalopride is a potent, selective and specific 5 HT(4) receptor agonist. As it is intended for treatment of intestinal motility disorders, it is important to note that prucalopride is devoid of anti cholinergic, anticholinesterase or nonspecific inhibitory activity and does not antagonise 5-HT(2A), 5-HT(2B) and 5-HT(3) receptors or motilin or CCK(1) receptors. PMID- 11438310 TI - Role of cysteinyl leukotrienes in nociceptive and inflammatory conditions in experimental animals. AB - The leukotrienes are potent inflammatory mediators, which may have a role in inflammatory diseases such as allergic rhinitis, inflammatory bowl disease and asthma. Zafirlukast, a cysteinyl leukotriene receptor antagonist, is claimed to be effective in asthma. However, it is not known whether these leukotrienes are involved in nociceptive and peripheral inflammation. The present study aimed to assess the role of cysteinyl leukotrienes in nociceptive and inflammatory conditions in experimental animals. Central nociception was assessed with tail flick and hot-plate methods and peripheral nociception was assessed by acetic acid-induced chemonociception in mice. Local administration (intraplantar) of carrageenan-induced hyperalgesia and inflammation, measured by paw withdrawal latency and paw volumes, respectively. Zafirlukast (2.5--20 mg/kg, p.o.) produced a significant and dose-dependent antinociceptive and antiinflammatory effect against acetic acid-induced chemonociception in mice and carrageenan-induced paw oedema in rats, respectively. Zafirlukast (2.5 and 5.0 mg/kg, p.o.) also attenuated the carrageenan-provoked hyperalgesia but did not alter the pain threshold in central nociception up to 20 mg/kg. Zafirlukast (5 and 10 mg/kg ) significantly inhibited exudate formation and migration of polymorphonuclear leukocytes in carrageenan-induced pleurisy. Further, zafirlukast (5 mg/kg) also reduced myeloperoxidase activity in carrageenan-treated paw. When nimesulide (2 mg/kg, p.o.) was co-administered with zafirlukast, the antinociceptive, antihyperalgesic and antiinflammatory effects of nimesulide were significantly increased as compared to the per se effect. The results indicate that cysteinyl leukotrienes are involved in nociceptive/inflammatory conditions. It is expected that combination of cysteinyl leukotriene receptor antagonist with cyclooxygenase inhibitor would prove to be a novel approach to treat complex inflammatory conditions. PMID- 11438312 TI - Role of cytokines in endometriosis. AB - OBJECTIVE: To review the literature on the role of cytokines in the pathogenesis of endometriosis and endometriosis-associated infertility. DESIGN: Pertinent studies were identified by a computer search of MEDLINE. References of selected articles were hand-searched for additional citations. RESULT(S): Recent studies suggest that the peritoneal fluid of women with endometriosis contains an increased number of activated macrophages that secrete various local products, such as growth factors and cytokines. Levels of several cytokines were reported to be elevated in the peritoneal fluid of women with endometriosis. Because the peritoneal environment may be controlled by locally regulated factors, cytokines are believed to play a role in the development and progression of endometriosis and endometriosis-associated infertility. A possible pathogenic mechanism links cytokines with endometriosis. CONCLUSION(S): Cytokines, which are produced by many cell types including endometriotic tissues, play diverse roles in the pathogenesis of endometriosis and endometriosis-associated infertility. More studies about the specific role of these cells and soluble factors are needed to improve understanding of endometriosis and to develop novel therapies. PMID- 11438313 TI - Bringing science communication into the new millennium. PMID- 11438314 TI - Effects of lower doses of conjugated equine estrogens and medroxyprogesterone acetate on plasma lipids and lipoproteins, coagulation factors, and carbohydrate metabolism. AB - OBJECTIVE: To determine the effects of lower doses of conjugated equine estrogens (CEE) alone or CEE and medroxyprogesterone acetate (MPA) on lipoproteins, carbohydrate metabolism, and coagulation/fibrinolytic factors. DESIGN: Randomized, double-blind, placebo-controlled study. SETTING: Multicenter substudy of the Women's HOPE trial. PATIENT(S): Seven hundred and forty-nine healthy, postmenopausal women. INTERVENTION(S): Women were randomized to receive the following doses in milligrams per day: 0.625 CEE; 0.625 CEE/2.5 MPA; 0.45 CEE; 0.45 CEE/2.5 MPA; 0.45 CEE/1.5 MPA; 0.3 CEE; 0.3 CEE/1.5 MPA; or placebo. MAIN OUTCOME MEASURE(S): Assessment of lipids, lipoproteins, glucose tolerance, and coagulation/fibrinolytic factors at baseline, cycle 6, and year 1. RESULT(S): One year of treatment with any of the CEE or CEE/MPA regimens studied increased high density lipoprotein cholesterol (HDL-C); the 10% increase in HDL-C for the CEE 0.45/MPA 1.5 group was similar to the CEE 0.625/MPA 2.5 group. Low-density lipoprotein cholesterol was significantly reduced in all of the active treatment groups except the CEE 0.3/MPA 1.5 group at cycle 13. Apolipoprotein A-I and triglyceride levels increased and apolipoprotein B levels decreased in all groups. The lipoprotein (a) level was reduced in the CEE 0.45/MPA 2.5, CEE 0.45/MPA 1.5, and CEE 0.625/MPA 2.5 groups. Minimal changes were observed in carbohydrate metabolism for all groups. Fibrinogen and PAI-1 activity decreased and plasminogen activity increased in all groups. Decreases in antithrombin III and protein S activities were significant for all active treatment groups except the CEE 0.3/MPA 1.5 group. CONCLUSION(S): Lower doses of CEE and CEE/MPA induce favorable changes in lipids, lipoproteins, and hemostatic factors with minimal changes in carbohydrate metabolism. PMID- 11438315 TI - Endometrial effects of lower doses of conjugated equine estrogens and medroxyprogesterone acetate. AB - OBJECTIVE: To determine the endometrial safety of lower doses of continuous combined conjugated equine estrogens (CEE) and medroxyprogesterone acetate (MPA). DESIGN: Randomized, double-blind, placebo-controlled study (the Women's Health, Osteoporosis, Progestin, Estrogen study). SETTING: Study centers across the United States. PATIENT(S): Healthy, postmenopausal women (n = 2,673) with an intact uterus. INTERVENTION(S): Patients received CEE 0.625 mg/day, CEE 0.625/MPA 2.5 mg/day, CEE 0.45 mg/day, CEE 0.45/MPA 2.5 mg/day, CEE 0.45/MPA 1.5 mg/day, CEE 0.3 mg/day, CEE 0.3/MPA 1.5 mg/day, or placebo for 1 year. Endometrial biopsies were evaluated at baseline, cycle 6, and year 1 using a centralized protocol. MAIN OUTCOME MEASURE(S): Efficacy of lower doses of CEE/MPA in reducing the incidence of endometrial hyperplasia rates associated with unopposed CEE. RESULT(S): Endometrial hyperplasia rates ranged from 0 to 0.37% for all CEE/MPA doses. Twenty-nine of the 32 cases of endometrial hyperplasia developed in women who were administered CEE 0.625 mg or CEE 0.45 mg. The incidence of endometrial hyperplasia increased with age for patients administered CEE alone. As expected, there were some inconsistencies among pathologists' ratings in the numbers of hyperplasias and incidence rates for the CEE-alone regimens. There were too few cases of hyperplasia in the combination groups to evaluate consistency among pathologists. CONCLUSION(S): One year of treatment with lower doses of CEE/MPA provides endometrial protection comparable to commonly prescribed doses. These regimens may be used by clinicians to individualize hormone replacement therapy in postmenopausal women. PMID- 11438316 TI - Pharmacokinetics of testosterone after percutaneous gel or buccal administration. AB - OBJECTIVE: To determine the pharmacokinetics of testosterone following its administration using transdermal gel or buccal lozenges. DESIGN: Pilot study. SETTING: University-based hospital. PATIENT(S): Ten bilaterally oophorectomized women. INTERVENTION(S): Daily micronized testosterone gel (1 mg) and testosterone propionate lozenge (1 mg). MAIN OUTCOME MEASURE(S): Total testosterone, androstenedione, dihydrotestosterone, 3alpha-androstanediol glucuronide, and sex hormone-binding globulin were measured in serum by specific radioimmunoassays; free testosterone levels were also calculated. RESULT(S): Before treatment, serum testosterone levels in the groups using the lozenge and gel were 16 +/- 4.0 and 20 +/- 6.0 ng/dL, respectively. Mean maximum testosterone levels obtained with the lozenge occurred 1 hour after administration on days 1 (692 +/- 236 ng/dL) and 14 (836 +/- 309 ng/dL) of treatment and fell precipitously thereafter. In contrast, testosterone levels obtained with the gel showed a prolonged rise reaching maximal levels of 97 +/- 78 and 100 +/- 60 ng/dL after 18 hours. The serum level patterns of free testosterone, dihydrotestosterone, and 3alpha androstanediol glucuronide were similar to the corresponding total testosterone levels. CONCLUSION(S): Administration of testosterone lozenge by buccal absorption produced a rapid and brief elevation of testosterone levels, with levels reaching upper limits of the male range. In contrast, transdermal testosterone gel absorption resulted in a prolonged elevation of testosterone levels, which were in the hyperandrogenic female range but resembled steady state pharmacokinetics. PMID- 11438317 TI - Effects of raloxifene treatment on uterine leiomyomas in postmenopausal women. AB - OBJECTIVE: To evaluate the effects of raloxifene administration on uterine and uterine leiomyoma sizes in postmenopausal women. DESIGN: Prospective randomized, double-blind, placebo-controlled clinical trial. SETTING: Department of Gynecology, Obstetrics, and Pathophysiology of Human Reproduction, University of Naples "Federico II", Italy. PATIENT(S): Seventy spontaneous postmenopausal women affected by uterine leiomyomas. INTERVENTION(S): Twelve cycles (of 28 days each) of treatment with raloxifene (60 mg daily per os) or placebo. MAIN OUTCOME MEASURE(S): At entry and at every 3 cycles, uterine and uterine leiomyoma dimensions were measured by means of transvaginal ultrasound. The difference between uterine and leiomyoma volumes (Delta size) was calculated in all subjects. The characteristics of uterine bleeding and the side effects of the treatments were assessed using a daily diary. RESULT(S): After 6, 9, and 12 cycles of therapy, in subjects treated with raloxifene, the mean uterine and uterine leiomyoma size were significantly decreased, and the mean Delta size significantly increased in comparison with basal values and the placebo group. No significant differences in uterine bleeding were detected between the two groups. CONCLUSION(S): In postmenopausal women raloxifene appears to act selectively on uterine leiomyomas, reducing their size. PMID- 11438318 TI - Quality of life in low-income menopausal women attending primary care clinics. AB - OBJECTIVE: To examine the quality of life and health status of a population of menopausal age primary care attendees that demographically has not been well represented in previous studies, and to describe the relationships between population characteristics and outcomes. DESIGN: Cross-sectional. SETTING: Community primary care clinics. PATIENT(S): Women 45-60 years of age within 5 years of their last period. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Participants provided demographic information and completed a series of questionnaires, including the Menopause Quality of Life Instrument (MENQOL) and the Short Form-36 health survey (SF-36). RESULT(S): Women who were employed, had higher levels of education, or higher levels of income reported better overall health and fewer menopausal symptoms. The study population scored significantly lower than the national norms on a summary survey of mental and physical health. There were no significant differences between ethnic groups with respect to either menopausal quality of life or health status. CONCLUSION(S): In a generally low income, poorly educated menopausal population, ethnicity did not significantly affect quality of life. Socioeconomic characteristics (less education and lower income) were associated with increased menopausal symptoms. PMID- 11438319 TI - Soluble ICAM-1 and E-selectin levels correlate with clinical and biological aspects of severe ovarian hyperstimulation syndrome. AB - OBJECTIVE: To assess the potential involvement of the soluble endothelial cell leukocyte adhesion molecules E-selectin and intercellular adhesion molecule-1 (ICAM-1) in the pathophysiology of capillary hyperpermeability in the ovarian hyperstimulation syndrome (OHSS). DESIGN: Controlled clinical study. SETTING: Women hospitalized for severe OHSS after ovulation induction for IVF at two tertiary medical centers. PATIENT(S): Eleven patients with severe OHSS undergoing IVF and 20 controls who had received a similar ovulation induction regimen and did not develop OHSS. INTERVENTION(S): Serial serum samples were obtained from all patients with OHSS from admission until discharge. Ascitic fluid was obtained from all patients by therapeutic paracentesis. Serum was obtained from all controls 4-8 days after ET. MAIN OUTCOME MEASURE(S): Samples were assayed for soluble E-selectin and soluble ICAM-1 by using enzyme-linked immunosorbent assay and results were correlated with clinical and biological aspects of OHSS. RESULT(S): Compared with controls, patients with severe OHSS had higher levels of soluble ICAM-1 and lower levels of soluble E-selectin detected in serum and ascites. Serum levels of soluble ICAM-1 decreased, while soluble E-selectin levels increased along with clinical and biological improvement. Serum soluble ICAM-1 showed significant positive correlation and serum soluble E-selectin showed significant negative correlation with clinical and biological aspects of severe OHSS. CONCLUSION(S): Soluble ICAM-1 and soluble E-selectin seem to be involved in the pathophysiology of capillary hyperpermeability in severe OHSS. PMID- 11438320 TI - Subcutaneously administered Repronex in oligoovulatory female patients undergoing ovulation induction is as effective and well tolerated as intramuscular human menopausal gonadotropin treatment. AB - OBJECTIVE: To determine the efficacy and safety of Repronex SC as compared with Repronex IM and Pergonal IM in patients undergoing ovulation induction. DESIGN: Randomized, open-label, multicenter, parallel group study. SETTING: Ten academic and private fertility clinics with expertise in ovualtion induction. PATIENT(S): Premenopausal anovulatory and oligoovulatory females (n = 115) undergoing ovulation induction. INTERVENTION(S): Down-regulation with leuprolide acetate followed by up to 12 days of treatment with gonadotropins and hCG administration and luteal phase progesterone support. MAIN OUTCOME MEASURE(S): Percentage of patients ovulating; percentage of cycles with follicular development meeting criteria for hCG administration; number of follicles recruited per cycle meeting hCG criteria; peak serum E(2) levels; rates of chemical, clinical and ongoing pregnancies; adverse events; injection-site pain scores. RESULT(S): There was no statistically significant difference in the percentage of women who ovulated among the treatment groups. However, Repronex SC was significantly more effective than Pergonal IM in producing follicular development in patients who met hCG criteria. There were no significant differences in clinical, ongoing, or continuing pregnancy rates or in multiple pregnancies among the treatment groups. No differences were found in the safety assessments, proportions or seriousness of adverse events or treatment discontinuations. Also, there were no differences between the three treatment groups in patient-recorded scores of injection-site pain or injection-site reactions. CONCLUSION(S): Repronex SC is as efficacious and well tolerated as Repronex IM or Pergonal IM in ovulation induction. Self administration of Repronex SC provides a convenient treatment alternative to daily IM injections. PMID- 11438321 TI - Recombinant human chorionic gonadotropin (rhCG) in assisted reproductive technology: results of a clinical trial comparing two doses of rhCG (Ovidrel) to urinary hCG (Profasi) for induction of final follicular maturation in in vitro fertilization-embryo transfer. AB - OBJECTIVE: To compare the efficacy and safety of 250 microg and 500 microg of recombinant hCG with 10,000 U USP of urinary hCG in assisted reproduction technology. DESIGN: Open, comparative, randomized, prospective clinical study. SETTING: Twenty tertiary care U.S. infertility centers. PATIENT(S): Two hundred ninety-seven ovulatory infertile women undergoing a single cycle of assisted reproduction technology. INTERVENTION(S): Patients were randomized 1:1:1 to 250 microg of recombinant hCG SC, 500 microg of recombinant hCG SC, or 10,000 U USP urinary hCG IM after completing gonadotropin stimulation. MAIN OUTCOME MEASURE(S): Number of oocytes retrieved per patient receiving hCG. Also, measures of oocyte maturity, embryo development, and luteal function, as well as pregnancy and pregnancy outcome. Adverse safety events, laboratory changes, local tolerance, and immunogenicity were also assessed. RESULT(S): Mean numbers of oocytes retrieved per treatment group were equivalent, 13.6, 14.6, and 13.7 with 250 microg of recombinant hCG, 500 microg of recombinant hCG, and urinary hCG, respectively. The numbers of 2PN fertilized oocytes on day 1 after oocyte retrieval, and 2PN or cleaved embryos on the day of embryo transfer, were significantly higher with 500 microg of recombinant hCG than with the lower dose. However, the incidence of adverse events also tended to be higher with this dose. CONCLUSION(S): Recombinant hCG is effective and well tolerated in the induction of final follicular maturation and luteinization in women undergoing assisted reproduction technology. Recombinant hCG (250 microg) SC is equivalent to 10,000 U USP of urinary hCG in this indication. PMID- 11438322 TI - Does increasing ovum donor compensation lead to differences in donor characteristics? AB - OBJECTIVE: To evaluate the effects of a compensation increase for anonymous ovum donors on demographic and social characteristics. DESIGN: Retrospective analysis. SETTING: The Mount Sinai Medical Center Ovum Donation Program. PATIENT(S): All program applicants for 2 years preceding (group I, n = 2,934) and 1 year following an increase in donor compensation (group II, n = 1,114; total N = 4,048). INTERVENTION(S): Compensation was increased from $2,500 to $5,000 per cycle. MAIN OUTCOME MEASURE(S): Demographic and social characteristics of applicants and donors. RESULT(S): More group II applicants (65.7%) than group I applicants (49.2%) returned an initial biographical questionnaire. Compensation level did not affect the percentage rejected at any stage in the application process or ultimately selected. There were no differences in donors in age, marital status, education, race, religion, or psychological profile. Group II donors had more previous pregnancies (group II mean = 1.2, group I mean = 0.6) and previous abortions (group II mean = 0.8, group I mean = 0.4). CONCLUSION(S): Increasing compensation may result in a higher percentage of potential donors completing an initial questionnaire but does not alter the demographic and social characteristics of selected donors. Adherence to a rigorous applicant screening ensures that donor characteristics remain independent from compensation. PMID- 11438323 TI - Comparison of implantation and pregnancy rates in African American and white women in an assisted reproductive technology practice. AB - OBJECTIVE: To compare IVF outcomes between infertile African American and white women. DESIGN: Retrospective cohort study. SETTING: Hospital-based IVF practice. PATIENT(S): Women undergoing IVF procedures between November 1996 and June 2000. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Implantation and pregnancy rates. RESULT(S): There were 24 African American and 273 white women < or =40 years of age who underwent 25 and 333 IVF cycles, respectively. African American women were more likely to have had tubal factor as a primary diagnosis, to have had a child, and to have undergone fewer previous assisted reproductive technology (ART) cycles as compared to white women. No differences between the two groups for clinical variables were noted with the exception of body mass index (BMI [kg/m(2)], 27.1 in African Americans vs. 24.8 in whites). Implantation rates were higher in African American than in white women (35% vs. 23%, respectively). Pregnancy rates were 71% in African Americans and 48% in whites. After adjustment for tubal factor, BMI, and parity, the odds ratio for pregnancy in African American women versus white women increased from 2.6 to 3.3. CONCLUSION(S): This is the first study to demonstrate a significantly higher clinical pregnancy rate in African American women as compared to white women undergoing ART. These data strongly contradict a recent study comparing the same two groups of women undergoing ART. We urge other ART centers to report their data pertaining to race. PMID- 11438324 TI - Immunological changes and stress are associated with different implantation rates in patients undergoing in vitro fertilization-embryo transfer. AB - OBJECTIVE: To evaluate the possible correlation between immunological changes and implantation rates in patients who undergo in vitro fertilization-embryo transfer (IVF-ET). DESIGN: Controlled clinical study. SETTING: University hospital. PATIENT(S): Forty infertile women undergoing IVF-ET. INTERVENTION(S): Stroop Color Word (CW) test, State-Trait Anxiety Inventory (STAI) test, blood sampling. MAIN OUTCOME MEASURE(S): Heart rate and systolic and diastolic blood pressure responses to Stroop CW; circulating T, B, T-helper (CD4), and T-suppressor (CD8) lymphocytes. RESULT(S): The total number of T lymphocytes increased significantly during superovulation, resulting in significantly higher levels in subjects achieving embryo implantation than in those showing a failure of implantation. An opposite trend was observed for the activated T cells. The number of T-helper lymphocytes and the T-helper/T-suppressor ratio showed a significant increase from baseline to the time of pick-up only in patients with implantation. CONCLUSION(S): A prolonged condition of stress, which causes a decreased ability to adapt and a transitory anxious state, is associated with high amounts of activated T cells in the peripheral blood. Such a condition, in turn, is associated with a reduced implantation rate in women undergoing IVF-ET. PMID- 11438325 TI - Factors useful in predicting the success of oocyte donation: a 3-year retrospective analysis. AB - OBJECTIVE: To establish prognostic relevance of parameters assessed in oocyte donation cycles. DESIGN: Retrospective analysis. SETTING: Large university-based donor oocyte program. PATIENT(S): All oocyte recipient cycles achieving embryo transfer from September 1995 to October 1998. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Pregnancy. RESULT(S): Recipient age and reproductive status, day 9 and 12 serum estradiol (E(2)) levels and a progesterone (P) level obtained 2 days after initiation of hormonal therapy did not correlate with pregnancy. Endometrial thickness, but not endometrial pattern, was useful in predicting pregnancy outcome. The clinical pregnancy and live-birth rate in cycles where the endometrial thickness was less than 8 mm was significantly lower when compared to cycles with an endometrial thickness > or =9 mm. Cycles where optimal quality embryos were transferred had the highest implantation (36%), clinical pregnancy (63%) and live birth (54%) rates and these rates were significantly higher than those of cycles where only poor quality embryos were available for transfer (10% implantation, 17% clinical pregnancy, and 8% live birth rates, respectively; P<.05). CONCLUSION(S): The most reliable predictive factors for pregnancy in oocyte donation cycles are the quality of the embryos transferred and the recipient's mid-cycle endometrial thickness. Recipient monitoring should minimally include ultrasound assessment of endometrial thickness. PMID- 11438326 TI - Concentrations of angiogenic factors in follicular fluid and oocyte-cumulus complex culture medium from women undergoing in vitro fertilization: association with oocyte maturity and fertilization. AB - OBJECTIVE: To determine the concentration of angiogenic factors (vascular endothelial growth factor [VEGF], basic fibroblast growth factor [bFGF], and angiogenin) in the follicular fluid (FF) and oocyte-cumulus complex culture medium (CM) of women undergoing IVF and to investigate the association of the concentrations with the maturity and fertilization of the oocyte. DESIGN: Prospective study. SETTING: Academic tertiary-care institution. PATIENT(S): IVF patients with unexplained or tubal factor infertility. INTERVENTION(S): Analysis of VEGF, bFGF, and angiogenin FF and CM concentrations. MAIN OUTCOME MEASURE(S): Oocyte maturity and fertilization and FF and CM angiogenic factor concentrations. RESULT(S): VEGF, bFGF, and angiogenin were determined in FF and CM. FF angiogenin concentrations were significantly higher when the oocyte was mature versus immature. CM VEGF concentrations were significantly higher when the oocyte was nonfertilized versus fertilized. Positive correlations were observed between angiogenic factors in CM. CONCLUSION(S): VEGF, bFGF, and angiogenin (determined for the first time) are secreted in the FF and CM. Elevated CM VEGF concentrations, probably implying oocyte-cumulus complex hypoxia, are negatively associated with oocyte fertilization. Elevated FF angiogenin concentrations are positively associated with oocyte maturity, possibly indicating angiogenin's biological role beyond neovascularization. PMID- 11438327 TI - Effect of a combined oral contraceptive containing 3 mg of drospirenone and 30 microg of ethinyl estradiol on the human endometrium. AB - OBJECTIVE: To provide an in-depth assessment of the effects of the combined oral contraceptive containing 30 microg of ethinyl estradiol and 3 mg of drospirenone (Yasmin, Schering AG, Berlin) on the endometrium by means of endometrial morphometry in comparison to an untreated cycle. DESIGN: The open, multicenter study consisted of one untreated precycle and 13 treatment cycles. SETTING: Four gynecologic clinics in Belgium, The Netherlands, and Switzerland were involved. PATIENT(S): Forty women with a history of regular menstrual cycles. INTERVENTION(S): Before the commencement of the trial, 3 months without any hormonal intake was obligatory. The first endometrial sample was done in the untreated precycle, adjusted to the day of LH peak plus 5 to 6 days. During the medication phase, endometrial samples were taken at cycle 3, 6 and 13. MAIN OUTCOME MEASURE(S): Primary outcome measure of the study was the morphologic assessment of the endometrium with measures such as glandular diameter, glandular epithelial height, and number of vacuolated cells per 1,000 glandular cells. Furthermore, the endometrial thickness was measured by ultrasound. RESULT(S): After 13 cycles of medication use the endometrium had an atrophic appearance in 63% of the subjects. The size of the glands, the glandular epithelial height, and the number of glands per square millimeter were already significantly reduced after 3 months' use. Histological and ultrasonographical evaluation of the endometrium indicated a suppression of the proliferative activity of the endometrium. CONCLUSION(S): The combination of 30 microg of ethinyl estradiol with 3 mg of drospirenone induces changes of the endometrium that are comparable with other combined oral contraceptives and exhibits a marked antiproliferative effect on the endometrium. The medication was proven to be an effective oral contraceptive and revealed good cycle control characteristics. PMID- 11438328 TI - Ovarian function during and after treatment with the new progestagen Org 30659. AB - OBJECTIVE: To evaluate ovarian function during 21 days of oral administration of different doses of Org 30659, a novel selective progestagenic steroid. DESIGN: Randomized, double-blind, dose-finding study. SETTINGS: Three centers in Austria, Sweden, and the United Kingdom. PARTICIPANTS: Eighty-one healthy women 19-40 years of age with regular ovulatory cycles. INTERVENTION: Daily oral administration of 0.060, 0.120, 0.180, or 0.240 mg of Org 30659, or 0.075 mg desogestrel (reference group), for 21 days. MAIN OUTCOME MEASURE(S): Once-daily measurements of follicular diameter and 17-beta estradiol, progesterone, FSH, and LH levels. RESULT(S): Daily treatment with Org 30659 for 21 days caused dose dependent suppression of ovarian activity. No ovulation was observed in any study group. On average, ovulation returned 16.5 to 22.1 days after treatment. The effects of desogestrel, 0.075 mg, were similar to those of 0.060 and 0.120 mg of Org 30659. All doses were well tolerated, as shown by the type of side effects that occurred, the absence of an effect on physical and laboratory findings, and the low rate of study discontinuation. CONCLUSION(S): Daily oral administration of 0.060-0.240 mg of Org 30659 suppresses ovarian function to a level sufficient to inhibit ovulation. This effect is dose-dependent, and the suppressive effect is readily reversible at all doses tested. Org 30659 can thus be safely administered orally for 21 days to healthy female volunteers in a dosage of 0.060 mg/d to 0.240 mg/d. PMID- 11438329 TI - Effects of trophoblast invasion on the distribution of leukocytes in uterine and tubal implantation sites. AB - OBJECTIVE: To distinguish endocrine and paracrine influences on leukocyte subpopulations at uterine and tubal implantation sites. DESIGN: Retrospective immunohistochemical study. SETTING: Departments of Anatomy, and Obstetrics and Gynecology, School of Medicine, RWTH University of Aachen, Aachen, Germany. PATIENT(S): Ten women with a viable ectopic pregnancy (EP), 25 women who had undergone elective first-trimester termination of pregnancy, and 4 women who had undergone hysterectomy with adnexectomy. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Quantitative analysis of leukocyte subpopulations at the implantation sites and their corresponding noninvaded tissues, decidual tissue from patients with EP, and tubal mucosa from normal menstrual cycle. RESULT(S): Similar numbers and characteristic distribution patterns of macrophages, T cells, and B cells were found at both normal intrauterine and tubal implantation sites. Natural killer (NK) cells were always absent from tubal mucosa. The number and distribution of leukocytes within decidual tissue from women with EP corresponded to those in the noninvaded decidual compartment in intrauterine pregnancy (IUP). CONCLUSION(S): Leukocyte populations present in the tubal and uterine mucosa are an intrinsic characteristic of these tissues. The distinct leukocyte distribution pattern at the implantation sites suggests that the invading trophoblast exerts a paracrine influence on endometrial and endosalpingeal leukocytes. The absence of natural killer cells from the tubal wall may be one reason for the higher degree of invasiveness of the trophoblast at the tubal implantation site. PMID- 11438330 TI - Determination of total menstrual blood loss. AB - OBJECTIVE: To develop and validate a simple method of measuring total menstrual blood loss using a pictorial representation of blood loss, the menstrual pictogram. DESIGN: A prospective evaluation of total menstrual blood loss measurement by the menstrual pictogram compared to the alkaline hematin technique. SETTING: Academic menorrhagia research clinic. PATIENT(S): One hundred twenty-one women; 62 women complaining of heavy menstrual blood loss, 59 women who considered their menstrual blood loss to be normal. INTERVENTION(S): Participants were asked to complete the menstrual pictogram through the period and collect their feminine hygiene products for an alkaline hematin assessment. MAIN OUTCOME MEASURE(S): Percentage agreement between blood loss measured by the gold standard alkaline hematin method and the menstrual pictogram. Extraneous blood loss was measured using a semiquantitative pictorial method. RESULT(S): The menstrual pictogram had a high level of agreement for blood collected on feminine hygiene products compared with the alkaline hematin method. Some women also lose a significantly large amount of extraneous blood, which is not proportional to the alkaline hematin blood loss assessment. CONCLUSION(S): The menstrual pictogram provides a simple means of measuring menstrual blood loss. It is no longer appropriate to ignore extraneous blood loss, particularly as there is no correlation between extraneous blood loss and that measured on feminine hygiene products. PMID- 11438331 TI - Use of human gametes obtained from anonymous donors for the production of human embryonic stem cell lines. AB - OBJECTIVE: To investigate the use of donated gametes for the production of human embryonic stem cell lines. DESIGN: Basic research study. SETTING: Assisted Reproductive Technology (ART) program at an academic institution. PATIENT(S): Consenting oocyte and sperm donors. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Oocytes were aspirated from oocyte donors (n = 12) and inseminated with frozen-thawed donor (n = 2) sperm followed by culture of embryos to day 5 or 6 in sequential media. The inner cell masses of expanded blastocysts were isolated using immunosurgery and cultured for 4-11 days on irradiated primary mouse embryonic fibroblasts (PMEFs). Viable cell colonies were passed every 7-10 days onto fresh PMEFs in the presence of leukemia inhibitory factor (0.1 microg/mL) and evaluated for appropriate cell surface markers. RESULT(S): Immunosurgery of 40 blastocysts resulted in the culture of 18 inner cell masses, which have produced three cell lines. One of these cell lines has been shown to stain positive for alkaline phosphatase and stage-specific embryonic antigen (SSEA)-4 and negative for SSEA-1, express telomerase activity, and produce hCG when allowed to differentiate. CONCLUSION(S): These findings demonstrate that the future production of human embryonic stem cell lines for therapeutic use is possible with the use of donated gametes. Many ethical issues were considered before the initiation of this study, and it was our goal to ensure that both oocyte and sperm donors understood the nature and purpose of the research before their participating in the study. PMID- 11438332 TI - Does response to treatment of ejaculatory duct obstruction in infertile men vary with pathology? AB - OBJECTIVE: To describe the pathology-specific response to transurethral resection of ejaculatory ducts (TURED) in patients with complete or partial ejaculatory duct obstruction and to evaluate the role of TURED in light of powerful assisted reproductive technologies. DESIGN: Retrospective clinical study. SETTING: University hospital. PATIENT(S): Thirty-eight infertile men with obstruction of the ejaculatory ducts. INTERVENTION(S): Diagnosis by transrectal ultrasonography or magnetic resonance imaging, and treatment with TURED. MAIN OUTCOME MEASURE(S): Changes in semen variables, pregnancy outcomes, and complication rates were analyzed before and after surgery. RESULT(S): Improvement in semen variables was significantly better in patients with partial obstruction (94%) of ducts than in those with complete obstruction (59%) (P=.04). Cystic obstruction, especially midline and eccentric cysts, responded best to TURED. Before surgery, all patients were candidates for IVF/ICSI; after surgery, 32% of azoospermic men and 81% of oligospermic men conceived spontaneously or were referred for IUI instead of IVF/ICSI. CONCLUSION(S): Ejaculatory duct obstruction due to cysts appears to respond best to TURED. In addition, TURED may decrease the need for IVF/ICSI as primary treatment in many cases. Finally, TURED may allow IVF/ICSI to be performed with ejaculated rather than surgically retrieved sperm. PMID- 11438333 TI - Microinjection of antisense c-mos oligonucleotides prevents the progression of meiosis in human and hamster oocytes. AB - OBJECTIVE: To investigate the role of c-mos proto-oncogene in the progression of meiosis in human and hamster oocytes. DESIGN: Controlled basic research study. SETTING: Assisted reproduction units at medical institutions. PATIENT(S): Consenting in vitro fertilization patients. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Maturation to metaphase II (MII) 24 hours following microinjection of prophase I (PI) hamster oocytes with antisense (AS) and sense (S) c-mos oligonucleotides. Control oocytes (C) injected with medium or left uninjected (UI). In human oocytes, maturation to metaphase II was also measured except culture was extended to 48 hours and the sense group was omitted. RESULT(S): The percentage of hamster oocytes reaching metaphase II after 24 hours was as follows: 1.5% (1 of 65) for the antisense group; 63.1% (41 of 65) in the sense group; 66.1% (41 of 62) in the control group; and 69.3% (52 of 75) in the uninjected group. The percentage of human oocytes at metaphase II was 33.3% (4 of 12) in the antisense group, 83.3% (10 of 12) in the control group, and 82.8% (24 of 29) in the uninjected group. CONCLUSION(S): These results demonstrate that injection of c-mos antisense oligonucleotide significantly inhibits the progression of meiosis in hamster (P=.0001) and human (P=.05) oocytes. Thus, c mos proto-oncogene may be one of the critical regulators of meiosis in these two species. PMID- 11438334 TI - Effect of activation with Ca ionophore A23187 and puromycin on the development of human oocytes that failed to fertilize after intracytoplasmic sperm injection. AB - OBJECTIVE: To determine the effect of sequential oocyte activation with calcium ionophore A23187 (A23187) and puromycin on oocytes that were unfertilized after intracytoplasmic sperm injection (ICSI). DESIGN: Laboratory examination. SETTING: The in vitro fertilization/embryo transfer unit in Tokushima University Hospital. PATIENT(S): Discarded unfertilized oocytes following ICSI. INTERVENTION(S): All 172 oocytes that showed no evidence of normal fertilization 18 hours after ICSI were exposed to A23187 (5 microM) for 5 minutes and consequently were treated with puromycin (10 microg/mL) for 5 hours. MAIN OUTCOME MEASURE(S): The activation rate, proportion of oocytes that showed two pronuclei with the second polar body (2PN2PB), and cleavage rate were calculated. Chromosomal analysis of the oocytes with 2PN2PB was also performed. RESULT(S): The treatment activated 146 out of 172 oocytes (84.9%) and 30.1% of the activated oocytes showed 2PN2PB. Sixteen of 25 oocytes with 2PN2PB (64.0%) cleaved. There was no difference in the activation rate, proportion of activated oocytes with 2PN2PB, or cleavage rate between oocytes that were injected with a motile spermatozoon and those injected with an immotile spermatozoon. Chromosomal analysis showed a normal diploid set of chromosomes (n = 46) in four of five analyzable oocytes. CONCLUSION(S): The sequential treatment of calcium ionophore A23187 and puromycin activates unfertilized oocytes after ICSI. The resultant oocytes with 2PN2PB can cleave. PMID- 11438335 TI - Expression of the alphav integrin adhesion molecule during development of preimplantation human embryos. AB - OBJECTIVE: To investigate the expression of alphav integrin in developing human embryos. DESIGN: Observational study. SETTING: University medical center. PATIENT(S): Women undergoing IVF cycles. INTERVENTION(S): Immunofluorescent monoclonal antibodies against alphav integrin were used to stain human preimplantation embryos. MAIN OUTCOME MEASURE(S): Immunofluorescence microscopy and computerized image analysis were used to evaluate both qualitative and quantitative expression of integrins in human embryos. RESULT(S): The alphav integrin subunit was found to be expressed in human embryos throughout their development (from two-cell stage up to blastocyst). The expression of alphav integrin subunit gradually increased throughout embryo development as measured quantitatively by image analysis. CONCLUSION(S): The expression of alphav integrin subunit throughout the development of preimplantation human embryos reinforces the concept of a role for integrins in the process of implantation. PMID- 11438336 TI - Human spermicidal activity of inorganic and organic oxidants. AB - OBJECTIVE: To identify prospective oxidants that rapidly immobilize human sperm upon contact with human semen. DESIGN: Inorganic, organic, and enzymatically generated oxidants were mixed with human semen and spermicidal activity was tracked by a modified Sander-Cramer assay. SETTING: Commercial and university based laboratories. PATIENT(S): Semen samples obtained through a university-based andrology laboratory. INTERVENTION(S): Not applicable. MAIN OUTCOME MEASURE(S): Quantitation of spermicidal activity of test oxidants. RESULT(S): Sperm lost motility within 20 seconds of exposure to enzymatically generated free iodine (I(2)). Toluidine blue, phenazine methosulfate, or methylene blue exhibited some, albeit much less, spermicidal activity. Oxidants formed by mixing ascorbic acid with Fe(III)-EDTA, xanthine with xanthine oxidase, or by exposing sperm to the nitric oxide generator, SIN-1 (3-morpholinosydnonimine hydrochloride), were far less potent spermicidal agents. CONCLUSION(S): Free I(2) formed in situ and presented to semen is an extremely potent spermicide. Additional studies on methods of generating de novo I(2) may be beneficial in developing a novel new class of nondetergent-based spermicides. PMID- 11438337 TI - Effect of endotoxin-induced reactive oxygen species on sperm motility. AB - OBJECTIVE: To define the mechanism of infection-induced damage of sperm. DESIGN: The effect of lipopolysaccharide (LPS)-induced reactive oxygen species (ROS) on sperm motility and its modification by scavengers were investigated. SETTING: Research laboratory of a university hospital. PATIENT(S): Normozoospermic semen samples were obtained from 37 healthy volunteers. INTERVENTION(S): The sperms were incubated in the presence of LPS with or without scavengers. MAIN OUTCOME MEASURE(S): Sperm motility was evaluated by a sperm quality analyzer (SQAIIB). ROS formation in semen samples was measured by a Berthold luminometer (LB953). RESULT(S): Motility of spermatozoa was decreased in the LPS-treated samples compared with that in the control groups. ROS was significantly higher in the LPS treated groups than in the control groups. The addition of ROS scavengers restored the motility index and suppressed ROS production in the LPS-treated semen samples. CONCLUSION(S): These data suggest that endotoxin-induced excessive production of ROS is responsible for the decrease in sperm motility and that antioxidant therapy may be a therapeutic option for infertile men with bacterial genital tract infection. PMID- 11438338 TI - Establishment of an animal embryo culture system containing various embryotropins and its efficacy for culturing ICR mouse one-cell embryos derived in vivo or in vitro. AB - OBJECTIVE: To develop an effective ICR mouse embryo culture medium. DESIGN: In vitro model study. SETTING: University-affiliated hospital. ANIMALS: Four-week old, superovulated mice. INTERVENTION(S): In vivo- or in vitro-derived one-cell embryos were cultured in preimplantation-1 medium (P-1). MAIN OUTCOME MEASURE(S): Preimplantation development. RESULT(S): In vivo-derived embryos were cultured in BSA-containing P-1, to which one of the following substances was added: [1] no addition, [2] amino acids (aa), [3] aa+hemoglobin (hb), [4] aa+hb+cysteine (cys), [5] aa+hb and glucose (glu) added at the four-cell, or [6] aa+hb and glu+cys added at the four-cell stage. More (P<0.05) blastocysts developed after aa or aa+hb addition than after no addition, and glu addition to such medium further stimulated the formation (54%). In P-1 with aa+glu, the addition of 1 microg/mL hb was optimal. Additional improvement of blastocyst formation (78%) was achieved by ethylenediaminetetraacetic acid (EDTA), supplementation and bovine serum albumin replacement with polyvinyl alcohol (PVA) did not inhibit the development. P-1 supplemented with aa, hb, glu, EDTA, and PVA also supported the development of in vitro-derived embryos (70%). CONCLUSION(S): A modified P-1 medium was developed, and it supported the development of both in vivo- and in vitro-derived ICR mouse embryos. PMID- 11438339 TI - Collagen membrane/fleece composite film reduces adhesions in the presence of bleeding in a rabbit uterine horn model. AB - OBJECTIVE: Determination of efficacy in presence of bleeding of CDS, a collagen/membrane fleece composite, in a rabbit uterine horn simple abrasion model. DESIGN: Randomized, controlled, and blinded study involving standard abrasion of the uterine horns with induction of moderate mesouterine bleeding. SETTING: Research laboratory. PATIENT(S): New Zealand White rabbits. INTERVENTION(S): No treatment (surgical control), CDS film, or INTERCEED barrier (negative reference control). MAIN OUTCOME MEASURE(S) AND RESULT(S): The extent (percent length uterine horn) with adhesions was assessed after 29 or 30 days. Adhesions formed in surgical controls to an extent (85.6% +/- 4.6%) consistent with historic data for this model. INTERCEED failed to reduce adhesions (78.1% +/ 7.7%) indicating that the test conditions of inadequate hemostasis were validated. CDS film, despite this inadequate hemostasis, reduced the extent of adhesions (31% +/- 7.4%; P<.01). Both the tenacity (P=.0008) and degree of uterine convolution (P=.000003) was reduced by CDS film but not by INTERCEED. CONCLUSION(S): Under conditions of inadequate hemostasis CDS effected a reduction in adhesion development. CDS may be useful adjuvant for procedures where hemostasis is difficult to achieve. PMID- 11438340 TI - Intrauterine insemination-ready versus conventional semen cryopreservation for donor insemination: a comparison of retrospective results and a prospective, randomized trial. AB - OBJECTIVE: To compare fecundity rates following intrauterine insemination (IUI) with donor sperm frozen conventionally versus an IUI-ready preparation. DESIGN: Both retrospective results and a prospective, randomized study where recipients were assigned to one of two sperm cryopreservation methods in each cycle of intrauterine insemination are reported. SETTING: University-based infertility practice, affiliated private practices, and andrology laboratory. PATIENT(S): Women desiring therapeutic insemination in an effort to establish pregnancy. INTERVENTION(S): Intrauterine insemination with donor sperm frozen conventionally or by an IUI-ready protocol. MAIN OUTCOME MEASURE(S): Cycle fecundity in donor IUI recipients. RESULT(S): In a retrospective analysis involving 642 inseminations in 209 recipients, 79 pregnancies were recorded for an overall pregnancy rate of 12.3% per insemination (or cycle): 11.3% with IUI-ready sperm and 13.9% with conventionally preserved sperm. In a follow-up prospective, randomized study, the pregnancy rate for IUI-ready sperm preparations was 36% per cycle (14 of 39) whereas that for conventionally preserved sperm was 19.6% per cycle (9 of 46). Thirteen of the 23 pregnancies occurred in the first study cycle of insemination; only two pregnancies were observed in patients undergoing more than four cycles of insemination. CONCLUSION(S): Cycle fecundity for IUI-ready donor sperm is equivalent to conventional cryopreserved sperm based on both prospective and retrospective assessments. PMID- 11438341 TI - Distribution of topical medication in the human vagina as imaged by magnetic resonance imaging. AB - OBJECTIVE: To assess the use of magnetic resonance imaging (MRI) to determine the varying distribution of a vaginally placed gel over time and with different levels of patient activity. DESIGN: Prospectives interventional trial. SETTING: University medical center. PATIENT(S): One nulliparous volunteer with normal menstrual cycles and no gynecologic disease who underwent seven MRI scans of the pelvis. INTERVENTION(S): Five mL of a commercially available topical spermicide was mixed with gadolinium-chelate magnetic resonance contrast material and introduced with a standard applicator. T1-weighted three-dimensional MRI was done to assess the distribution of the gel. MAIN OUTCOME MEASURE(S): Gel thickness and distribution. RESULT(S): The initial bolus of gel was delivered into the upper portion of the vagina, above the urogenital diaphragm. Thereafter, it spread into the vaginal fornices and "flattened" to cover the lateral aspects of the vagina. Without ambulation, the majority of spread was confined to the upper vagina. With ambulation and longer elapsed time, the gel spread further in the upper vagina and into the lower vagina, and significant vaginal surface coverage increased significantly. CONCLUSION(S): Magnetic resonance imaging can be used to monitor the spread of vaginally placed products and to evaluate coverage of topical drugs used for prevention and treatment, including those used for HIV prophylaxis. PMID- 11438342 TI - Acrosome components after intracytoplasmic sperm injection: the decondensation frontier. PMID- 11438343 TI - A case of forearm amputation after ovarian stimulation for in vitro fertilization embryo transfer. AB - OBJECTIVE: To report a case of forearm amputation after ovarian stimulation for IVF-ET. DESIGN: Case report. SETTING: A university hospital. PATIENT(S): A 41 year-old woman, who had coagulation disorder as a result of an ovarian hyperstimulation syndrome (OHSS) for IVF-ET. INTERVENTION(S): Retrospective evaluation of angiographic studies and surgical treatment. MAIN OUTCOME MEASURE(S): Medical follow-up after forearm amputation due to OHSS. RESULT(S): The patient underwent many cycles of IVF-ET with administration of purified FSH (75 IU 10 times per day, for 12 days) and chorionic gonadotropin (5,000 IU). The patient had a coagulation disorder as a result of OHSS, with thrombosis of the axillary vein, recurring after thromboarterectomy and leading to the paradoxical result of the amputation of an arm. CONCLUSION(S): An ethical evaluation of this case is mandatory, since the desire for pregnancy, the role of medical science, health, and human life itself are all factors involved. PMID- 11438344 TI - Inherited thrombophilia and in vitro fertilization implantation failure. PMID- 11438345 TI - Low-dose aspirin is effective for treatment of recurrent miscarriage in patients with decreased coagulation factor XII. PMID- 11438346 TI - Blood loss following noncomplicated transvaginal oocyte retrieval for in vitro fertilization. PMID- 11438347 TI - A 10-year follow-up study of 1,086 cases of nonsurgical reversible vas occlusion. PMID- 11438348 TI - Reproductive outcome according to semen quality in couples with infertility problems. PMID- 11438349 TI - Social stress. Acute and long-term effects on physiology and behavior. PMID- 11438350 TI - Primary social relationships influence the development of the hypothalamic- pituitary--adrenal axis in the rat. AB - In the developing rodent, there is a period from about days 4 to 14 when the adrenal response to stress is either minimal or non-existent. This has been designated as the stress hyporesponsive period (SHRP). Numerous experiments have demonstrated that maternal factors are critical for the regulation of the pup's hypothalamic--pituitary--adrenal (HPA) axis and the maintenance of the SHRP. Following 24 h of maternal deprivation, the neonatal rat shows elevated basal levels of corticosterone (CORT) and exhibits a robust CORT and ACTH response to mild stress. Further, c-fos mRNA in the paraventricular nucleus (PVN) is enhanced following stress in deprived pups. At least three aspects of maternal behavior play a role in the regulation of the HPA axis during development. Tactile stimulation appears capable of inhibiting most of the brain-related changes that occur following maternal deprivation. Feeding is essential for maintaining the adrenal-unresponsive and reduces the sensitivity of the adrenal to ACTH. Passive contact suppresses the response to stress. In the adult, corticotropin-releasing hormone (CRH) is the major neuropeptide that controls pituitary ACTH secretion. In the maternally deprived pup, CRH gene transcription is downregulated and arginine vasopressin (AVP) appears to assume the major regulatory hormone that modulates ACTH. These data all indicate that maternal factors are responsible for actively inhibiting the endocrine responses to stress postnatally. Thus, during development, most of the peripheral and central stress-responsive systems are capable of being activated. However, under conditions of normal dam-pup interactions, these responses are mostly suppressed by the dam's behavioral interaction with the pups. PMID- 11438351 TI - Animal models of social stress: effects on behavior and brain neurochemical systems. AB - Social interactions serve as an evolutionarily important source of stress, and one that is virtually ubiquitous among mammalian species. Animal models of social stress are varied, ranging from a focus on acute, intermittent, or chronic exposure involving agonistic behavior, to social isolation. The relative stressfulness of these experiences may depend on the species, sex, and age of the subjects, and subject sex also appears to influence the value of hypothalamic- pituitary--adrenal (HPA) axis activity as a general criterion for stress response: higher glucocorticoid levels are typically found in dominant females in some species. Social stress models often produce victorious and defeated, or dominant and subordinate, animals that may be compared to each other or to controls, but the appropriateness of specific types of comparisons and the interpretations of their differences may vary for the different models. Social stress strongly impacts behavior, generally reducing aggression and enhancing defensiveness, both inside and outside the stress situation. Social and sexual behaviors may be reduced in subordinate animals, as is activity and responsivity to normally rewarding events. However, some components of these changes may be dependent on the presence of a dominant, rather than representing a longer-term and general alteration in behavior. Social stress effects on brain neurotransmitter systems have been most extensively investigated, and most often found in serotonin and noradrenergic systems, with changes also reported for other monoamine and for peptidergic systems. Morphological changes and alterations of neogenesis and of cell survival particularly involving the hippocampus and dentate gyrus have been reported with severe social stress, as have longer-term changes in HPA axis functioning. These findings indicate that social stress models can provide high magnitude and appropriate stressors for research, but additionally suggest a need for caution in interpretation of the findings of these models and care in analysis of their underlying mechanisms. PMID- 11438352 TI - Topographic architecture of stress-related pathways targeting the noradrenergic locus coeruleus. AB - Peripheral sympathetic nerves and brainstem noradrenergic neurons of the locus coeruleus (LC) respond in parallel to a variety of stress-related stimuli which results in norepinephrine release both peripherally and centrally. Elucidation of central pathways subserving modulation of LC neurons point to extranuclear noradrenergic dendrites of LC somata that extend into peri-coerulear areas as a major target of afferents that participate in behavioral and physiological responses to stress. Anterograde tract tracing combined with immunoelectron microscopic detection of the catecholamine synthesizing enzyme tyrosine hydroxylase (TH) has demonstrated that the nucleus of the solitary tract (NTS) and the ventrolateral aspect of the periaqueductal gray (PAG), regions that participate in coordinating autonomic and motor behavior in response to stress, preferentially target the rostral ventromedial aspect of the peri-LC. In contrast, limbic forebrain afferents including the central nucleus of the amygdala (CNA) and the bed nucleus of the stria terminalis (BNST), regions that coordinate emotional responses to external stressors, provide direct synaptic input to noradrenergic dendrites that extend into rostral dorsolateral peri coerulear areas. Neurochemical identification of transmitter systems impinging on LC indicate that the CNA provides corticotropin-releasing factor (CRF), a peptide essential for integrated physiological responses to stress, to the dorsolateral LC. Endogenous opioid peptides that originate from medullary sources, however, target primarily the "core" of the LC. Our physiological data suggest that stress engages CRF and opioid afferents to the LC, which have opposing influences on this noradrenergic system. The balance between opioid and CRF influences acting in the LC may, in part, maintain the balance of active and passive coping behaviors in response to stress. Understanding the afferent and neurochemical organization of the LC may help elucidate adaptations in neural circuits associated with stress which impact on central noradrenergic function. PMID- 11438353 TI - Psychosocial stress, glucocorticoids, and structural alterations in the tree shrew hippocampus. AB - Animal models for chronic stress represent an indispensable preclinical approach to human pathology since clinical data point to a major role of psychological stress experiences, acute and/or chronic, to the development of behavioral and physiological disturbances. Chronic emotional arousal is a consequence of various types of social interactions, and one major neurohumoral accompaniment is the activation of the classic stress circuit, the limbic--hypothalamic--pituitary- adrenocortical (LHPA) axis. The adrenocortical glucocorticoid hormones cortisol and corticosterone are principal effectors within this circuit since they affect neurotransmission and neuroendocrine control, thus having profound effects on mood and behavior. Using the experimental paradigm of chronic psychosocial stress in tree shrews, we investigated the impact of aversive chronic social encounters on hippocampal structure and function. In chronically stressed animals, we observed dendritic atrophy of hippocampal pyramidal neurons and an impairment of neurogenesis in the dentate gyrus. However, a stress-induced loss of hippocampal neurons was not observed in this animal model. This review summarizes our recent results on structural changes occurring during chronic stress in neurons of the hippocampus and their potential influence on learning and memory. We discuss whether these changes are reversible and to what extent glucocorticoids might be responsible for the stress-induced effects. PMID- 11438354 TI - Chronic subordination stress in male tree shrews: replacement of testosterone affects behavior and central alpha(2)-adrenoceptors. AB - Subordination stress induced by social defeat in male animals is known to inhibit gonadal functions and it has been discussed whether the resulting deficit in testosterone might play a role in subordination behavior. One of the major transmitter systems involved in regulation of behavior is the noradrenergic system. To analyze whether a testosterone replacement can alter subordination behavior and whether this might be related to changes in the brain noradrenergic system, we quantified alpha(2)-adrenoceptors (alpha(2)-ARs) in the central nervous system of male tree shrews. Animals were submitted to chronic subordination stress and received testosterone at the same time. Behavior was monitored during all phases of the experiment: the control period of 10 days, the period of social stress lasting 10 days when subordinates were confronted daily with a dominant male, and, subsequently, the stress and treatment period of 18 days when in parallel to the stress, animals received either injections of testosterone or vehicle. Brain alpha(2)-ARs were quantified by in vitro receptor autoradiography using the antagonist ligand (3)H-RX821002. Locomotor activity decreased significantly during the stress period and was not re-normalized by testosterone. In contrast, testosterone re-normalized scent marking behavior and autogrooming, parameters that had both been reduced due to the subordination stress. Vehicle injections improved none of these behaviors. In 8 of 10 brain regions that were analyzed, numbers of alpha(2)-adrenergic binding sites were increased in stressed animals that received vehicle injections, but a difference between testosterone and vehicle injected animals was only observed in five regions. These brain regions are all known to be involved in emotional behavior (anterior hypothalamus, medial nucleus of the amygdala, cingulate cortex) or autonomic regulation, respectively (solitary tract nucleus, dorsal motor nucleus of vagus). Therefore, our data show that testosterone influences behavior of male subordinates and modulates alpha(2)-AR expression in their brains. Androgen mediated alterations in receptors occur in brain regions that are known to be involved in emotionality, e.g., in the anterior hypothalamus which regulates aggressive behavior. One can therefore conclude that alpha(2)-ARs contribute to neuronal functions that are responsible for subordination of stress behavior, and that testosterone-induced receptor changes are related to the partial restoration of normal behavior. PMID- 11438355 TI - Persistent suppression of ethanol self-administration by brief social stress in rats and increased startle response as index of withdrawal. AB - Excessive alcohol drinking is often linked to the experience of stress, but experimental approaches using animal models of alcohol self-administration have had widely varying outcomes. The objective was to determine how daily exposure to brief, predictable social stress would change alcohol self-administration in rats in a daily limited access protocol. Male Long-Evans rats had either access to a 10% ethanol solution for 15 min in the home cage setting (n=20) or were reinforced with 15% ethanol deliveries for every fifth lever press (n=10). Subsequently, all rats were subjected to brief social stress for five consecutive days. Social stress consisted of attacks by an opponent for 5 min followed by exposure to threats while in a protective cage for 30 min. In both the home cage drinking and operant conditioning groups, social stress exposure significantly decreased alcohol intake or rate of alcohol reinforcements, respectively. When alcohol intake was scheduled immediately before social stress (i.e., 24 h after the previous social stress episode), a decrease was observed with a delay of 1 or 2 days. When alcohol intake was scheduled 4 h after stress, no changes in intake or alcohol reinforcements were observed. Animals that consumed a low dose of ethanol displayed less defensive behavior during social stress compared to water drinking animals, and showed an increased startle reflex at 8 and 56 h after discontinuation of daily ethanol access. The current experimental protocols of social defeat stress reveal a transient suppression rather than a facilitation of alcohol consumption. PMID- 11438356 TI - Psychosocial vs. "physical" stress situations in rodents and humans: role of neurotrophins. AB - Nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are well studied polypeptide growth factors involved in the development and maintenance of specific peripheral and central populations of neuronal cells. In addition to its role as a neurotrophic agent, NGF controls very complex functions in vertebrate physiology. A variety of cells outside the nervous system are in fact able to synthesize NGF including epithelial cells, fibroblasts, lymphocytes, and macrophages. NGF target cells have been identified in the nervous, immune, and endocrine systems, suggesting that NGF may operate through multiple paths to ultimately regulate physiological homeostasis and behavioral coping. We used a mouse model of social stress to demonstrate that NGF levels increase both in plasma and in the hypothalamus following intermale aggressive interactions. The investigation has been extended to other species, including humans, to show that labour, lactation, and the anticipation of the first jump with a parachute also result in increased NGF plasma levels and in changes in the distribution of NGF receptors on lymphocytes. BDNF activation is caused by both physical and social stress events. The aim of this review is to (1) outline the current understanding of the roles of NGF and BDNF in stress-related physiological changes in vertebrates, in particular for physical vs. psychological stressors, which may activate both similar and different neurobiological pathways, and (2) summarize recent efforts to derive pharmacological strategies from the increasing body of BDNF and NGF neurobehavioral data. PMID- 11438357 TI - Role of environmental factors on brain development and nerve growth factor expression. AB - Numerous evidences suggest that early life events can affect the development of the nervous system, contributing in shaping interindividual differences in vulnerability to stress or psychopathology. A number of studies have shown that mothering style in rodents can produce neuroendocrine, neurochemical, and behavioral changes in the adult, although the basic mechanisms initiating this cascade of events still need to be investigated. This paper reviews research performed in our and other laboratories investigating some of the features characterizing hypothalamic--pituitary--adrenal (HPA) axis activity of rodents during early development, with a special emphasis on extrinsic, social regulatory factors, such as the mother and the siblings. In addition, a possible role for neurotrophins as mediators of the effects of external manipulations on brain development is suggested. PMID- 11438358 TI - A social conflict increases EEG slow-wave activity during subsequent sleep. AB - Electroencephalogram (EEG) slow-wave activity (SWA) during non-rapid eye movement (NREM) sleep is widely viewed as an indicator of sleep debt and sleep intensity. In a previous study, we reported a strong increase in SWA during NREM sleep after a social conflict in rats. To test whether this increase in SWA reflects normal physiological sleep or an unrelated by-product of the stress, we now measured the effect of a conflict in combination with extended sleep deprivation by means of gentle handling. We anticipated that if the social defeat-induced SWA reflects a true sleep debt, the drive for it would persist during the extended wakefulness. Male rats were subjected to a 1-h social conflict followed by 5 h of sleep deprivation by gentle handling or to 6 h gentle handling alone. The manipulations took place during the second half of the dark phase and recovery sleep was recorded during the subsequent light phase. Neither of the two procedures caused a significant change in the total duration of NREM or REM sleep thereafter. Yet, both modes of sleep deprivation induced a strong increase in SWA during NREM sleep. This SWA was significantly higher for 6 h after sleep deprivation consisting of a social conflict followed by gentle handling, as compared to sleep deprivation by handling alone. Thus, the SWA increasing effect of the conflict persisted during the extended wakefulness. The data confirm that social defeat stress accelerates the build up of sleep debt and support the notion that sleep debt and subsequent NREM sleep intensity not only depend on the duration of prior wakefulness but also on what animals experience during that waking. PMID- 11438359 TI - The link between stress and the efficacy of anxiolytics. A new avenue of research. AB - There is an important discrepancy between animal models of anxiety and human anxiety patients: while experimental animals are usually unstressed, patients usually have a long history of stress. Stressful life events not only contribute to the development and/or maintenance of mood disorders in humans but also affect the efficacy of anxiolytic treatment in both animals and humans. The effects are not trivial: sometimes stress may totally abolish the effects of certain anxiolytics. Therefore, the relationship between stress exposure and anxiolytic efficacy should be an important area of research and drug development. There are several benefits that may be derived from similar studies: (i) the effect of stress may unravel yet unknown aspects of the mechanism of action of different drugs; (ii) the relevance of laboratory studies would increase; and (iii) laboratory findings may provide cues regarding the clinical use of drugs. PMID- 11438360 TI - Cardiac autonomic responses to intermittent social conflict in rats. AB - Intermittent exposure to the same stressor can lead to a gradual decline in physiological, neuroendocrine and behavioral stress responses (habituation). We investigated possible habituation of cardiac autonomic responsiveness and susceptibility to cardiac arrhythmias in male rats exposed to either intermittent social victory (VIC) or defeat (DEF) stress (10 exposures in each case). Electrocardiograms were recorded via radiotelemetry and the sympathovagal balance at the level of the heart was evaluated via time-domain measurements of heart rate variability, namely average R--R interval (average time interval between two consecutive heart beats, RR), the standard deviation of RR (SD(RR)) and the root mean-square of successive R--R interval differences (r-MSSD). Values of these parameters were significantly lower in DEF as compared to VIC rats in the second part of the test period (from Minute 6 to Minute 15), suggesting a more pronounced sympathetic dominance in the former group of animals. Accordingly, the occurrence of the most frequent cardiac arrhythmias (ventricular and supraventricular premature beats) was higher in DEF rats. Habituation of cardiac autonomic responsivity was observed across repeated exposure to victory, both in terms of sympathovagal balance and susceptibility to cardiac tachyarrhythmias, whereas no habituation was found in repeatedly defeated animals. A possible explanation to this discrepancy could be the different degree of controllability characterizing the two social challenging situations. PMID- 11438361 TI - Social stress, myocardial damage and arrhythmias in rats with cardiac hypertrophy. AB - In rat models of cardiac hypertrophy (moderate aortic coarctation: ACm, n=18; severe aortic coarctation: ACs, n=27; aging: OLD, n=25; spontaneous chronic hypertension: SHR, n=18) and properly matched control animals (C(ACm), n=17; C(ACs), n=19; C(OLD), n=24; C(SHR), n=22), we investigated the relative contribution of intense autonomic activity and cardiac structural damage to ventricular arrhythmogenesis. We used an "in vivo" to tissue level approach, by correlating in the same animal: (i) social stress-induced ventricular arrhythmias, telemetrically recorded, and (ii) left ventricular weights (LVW) and amount and geometrical properties of myocardial fibrosis (MF). Arterial blood pressure was significantly higher in ACm (+11%), ACs (+28%) and SHR (+34%) than in controls. LVW were approximately 20% greater in ACm, ACs and OLD and 50% greater in SHR. MF was about twice as great and characterized by more frequent occurrence of microscopic scarring in ACm and ACs, and eight times greater and associated with both a higher number and a larger size of fibrotic foci in OLD and SHR compared to controls. Social stress increased ventricular arrhythmia vulnerability in all models of cardiac hypertrophy, as well as in controls. The arrhythmogenic action of stress was facilitated in ACs, OLD and SHR. A correlation between structural cardiac remodeling and ventricular arrhythmias was found only in SHR and OLD, which exhibited the greatest increase in LVW and/or MF. Social stress proved to be a valuable tool for analyzing the combined effects of autonomic stimulation and altered myocardial substrate on the genesis of potentially life-threatening arrhythmias in social animals. PMID- 11438362 TI - A link between psychosocial factors and blood pressure trend in women. AB - The powerful effect of psychosocial and acculturating influences on population blood pressure trends seems to be confirmed, through longitudinal observations, in the nuns in a secluded order. After initial observation had been made on culture, body form, blood pressure, diet, and other variables in 144 nuns and 138 laywomen, included as a control group, a 32-year follow-up study was undertaken. Most striking were opposite trends noted between the two groups in blood pressure trend. During the follow-up period blood pressure remained remarkably stable among the nuns. None showed a rise in diastolic blood pressure to above 90 mmHg. By contrast the control women showed the expected increase in blood pressure with age. This resulted in a gradually greater difference (Delta>30/15 mmHg) in systolic and diastolic blood pressure between the two groups, which was statistically significant. Fatal and nonfatal events were exceedingly lower in the nuns than in the control women over the follow-up period. It appears reasonable to attribute much of the difference in blood pressure to the different burden in psychosocial factor and to the preserved peaceful lifestyle of the nuns. PMID- 11438363 TI - Social stress in anorexia nervosa: a review of immuno-endocrine relationships. AB - Anorexia nervosa (AN) patients have difficulty in establishing social contacts, leading to tension, anxiety and full-blown stress reactions. Stress hormones are chronically increased in AN, while immune function, which is involved in physical and psychological coping capacities, is mostly unimpaired. We examined immune function in a group of anorexics by measuring the T-lymphocyte proliferative response to stimulation with phytohemagglutinin (PHA), before and after in vivo acute administration of corticotropin-releasing hormone (CRH), to mirror a stress reaction. The responses of anorexics, before and after CRH stimulation, did not differ from those of controls. In a second group of anorexics, we measured plasma concentrations of interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) before and after psychopharmacological (fluoxetine, amineptine) therapy. Basal values of the cytokines were not different in patients and controls, and did not change during therapy. In the same patients, we measured basal concentrations of soluble IL-1 beta receptor antagonist (s-IL-1 beta-RA), soluble IL-6 receptor (sIL-6-R) and soluble TNF alpha receptors I and II (sTNF-alpha-R-I and -II). S-IL-1 beta-RA and sTNF-alpha R-I and -II levels were not different in patients and controls, while those of s IL-6-R were lower than normal in anorexics. The normality of most of the immune parameters in our anorexics, in basal conditions, after a stressful stimulation and after pharmacological manipulation of neurotransmitters suggests that the well-known interrelation among immune, neuroendocrine and central nervous system functions is not maintained in AN, the immune system being somehow unresponsive to stimuli. PMID- 11438364 TI - Behavioral and physiological responses to stress are affected by high-fat feeding in male rats. AB - Interactions between monoaminergic neurochemistry and macronutrient intake have been frequently shown. Because monoaminergic systems in the brain are also closely involved in behavioral and physiological stress responses it can be hypothesized that differences in the macronutrient composition of diets are reflected in these responses. The present studies, therefore, were designed to assess the consequences of a change in dietary macronutrient composition on a variety of physiological and behavioral responses (both acute and long-term) to a number of stressors. The effect of chronic high-fat (HF; 61% kcal from fat) feeding on the stress responses was compared with controls receiving regular high carbohydrate (HC; 63% kcal from carbohydrates) laboratory chow. Rats were kept on this diet for at least 2 months before they were exposed to either psychological (social defeat) or physiological (lipopolysaccharide, LPS, administration) stress. At baseline, chronic HF feeding caused a slight, but significantly reduction in body temperature relative to that observed in HC-fed rats. Following social defeat or LPS injection, HF feeding caused a faster recovery of the body temperature increase relative to animals on the HC diet. Stress-induced suppression of home cage locomotor activity and body weight gain were also reduced by HF feeding. The serotonergic 5-HT(1a) receptor hyposensitivity that was observed in HC-fed rats 2 weeks after stress was absent in the HF regimen. Although the present results cannot be readily interpreted as showing purely beneficial effects of high-fat diets on stress responsivity, the findings in the present study do encourage further investigation of possible ameliorating effects of high-fat diets on aspects of the behavioral and physiological response stress. PMID- 11438365 TI - Intrinsic and environmental influences on immune senescence in the aged monkey. AB - Several factors associated with the age-related decline in immunity were examined in three studies on aged rhesus monkeys. Natural killer (NK) cell activity was found to be low in many monkeys after 20 years of age, but exceptionally long lived animals, older than 25 years, often had vigorous cytolytic responses. When NK activity was decreased in an aged monkey, it was predictive of fewer years of survival and a younger age at death. This prediction of mortality was associated with one nonimmune biomarker of aging in the monkey: nail growth rate. Monkeys with very slow nail growth and low NK activity were likely to die sooner. Although these findings might suggest an immutable course for the aging process, the housing conditions of old monkeys also had a pronounced effect on their NK activity. The highest NK responses were found in old monkeys housed with just one other old animal when compared to living alone or with just a young, juvenile monkey. It remains to be determined whether this type of psychosocial influence could have a sustained effect on immunity and ultimately change the pace of aging and time to mortality. PMID- 11438366 TI - Social stress in laboratory rats: behavior, immune function, and tumor metastasis. AB - This report summarizes data from social confrontations studies in laboratory rats dealing with the effects of psychosocial stress on immune functioning and tumor metastasis. The paper focuses on the physiological alterations observed in subdominant males after 2 days of continuous social confrontation. A significant loss of body mass and elevated plasma concentrations of adrenal hormones in subdominant males indicate a stressful social environment. Subdominant males showed lower numbers of blood CD4 and CD8 T cells as well as reduced activity levels of T cells and natural killer (NK) cells relative to control subjects. In order to evaluate the possible health impact of suppressed NK functioning, we used the MADB 106 tumor model. A 10-fold lower tumor clearance in subdominant males demonstrates suppression of the animals' capacity to prevent metastatic development. The relationship between individual behavior and immunological outcome is briefly discussed. Together, the study of male rats in social confrontations appears to be a good model to investigate stress-induced immune modulation and tumor metastasis under relatively naturalistic social conditions. PMID- 11438367 TI - delta-Opioid modulation of social interactions in juvenile mice weaned at different ages. AB - The environmental stimulus of weaning has been shown to affect both the developmental expression of social behavior and the maturation of the opioid delta-receptors' subpopulation in altricial rodents. The aim of this study was to address both these issues by using the social interaction paradigm. Separate groups of male and female mice were randomly assigned to three different weaning ages -- early (Wean-15), regular (Wean-20), and delayed (Wean-25) -- and assessed when 30 days old under intraperitoneal administration of the selective delta opioid agonist SNC80 (0, 0.1, or 0.3 mg/kg). Wean-15 male and female subjects were much more involved in investigating the partner as well as the cage environment compared to the regular Wean-20 group. An increased social investigation was also found as a consequence of delayed weaning in the female group. The neurobehavioral changes induced by the manipulation of weaning age were also reflected in an altered responsivity to the effects of SNC80 administration. The drug-induced increase in the expression of investigative and affiliative social interactions was further magnified by early weaning. A delayed weaning time was instead associated with reduced sensitivity to the drug, which suggests a delayed maturation of the system. As a whole, the present results indicate that the time of weaning is able to markedly affect the expression of social interactions of adolescent mice by possibly exerting a direct modulatory role on the development of the still plastic delta-opioid system. PMID- 11438368 TI - Social status in mice: behavioral, endocrine and immune changes are context dependent. AB - The aim of the present study was to investigate the effect of social status on the endocrine, immune and behavior response of male mice. We found that in mice reared in a group of siblings since weaning, no difference exists between dominants and subordinates in basal corticosterone level, in behavior in the open field test (OFT) and in a series of immune parameters. These results suggest that living with siblings is not a stressful condition for either dominant or subordinate mice. Therefore, group-housed siblings can be regarded as a valid control group in social stress studies. When mice were subjected to chronic psychosocial stress for 21 days, four types of social outcome occurred: residents becoming dominants, intruders becoming subordinates, residents becoming subordinates and intruders becoming dominants. Interestingly, the behavioral profile in the OFT revealed a status-dependent effect, with resident dominants (RD) and intruder dominants (InD) showing the highest locomotor and exploratory activity, whereas the corticosterone level was higher than control for all four categories. In addition, a context-dependent effect emerged at the immune level: resident subordinates (RS) had a reduced splenocyte proliferation and IL-4 and IL 10 production. Mice in all the other three social ranks showed no immune alterations. Therefore, the loss of an individual's social rank position seems a promising field of study to investigate the psychological impact of stressful events. PMID- 11438369 TI - Social stress in mice: gender differences and effects of estrous cycle and social dominance. AB - A large discrepancy in the possibility of inducing social stress in the two genders exists. Since generalizations of findings from one sex to the other appear not to be valid, reliable models of social stress in females are needed. We examined the effects of social context in the housing environment, as a possible source of stress, on exploration and anxiety in male and female mice, taking into account the estrous phase for females and the social status for males as additional variables. Mice housed individually or with siblings were tested in a free-exploratory paradigm of anxiety (where test animals have a choice to stay in their home cage or to explore an open field, OF). Individually housed females did not leave their home cage for long periods, explored less the unfamiliar area and displayed higher risk assessment, a behavioral profile suggestive of lower propensity for exploration and higher level of anxiety compared with group-housed females. Individually housed males tended to show an opposite profile. Proestrus mice were less sensitive to the decrease of exploratory propensity induced by individually housing compared to estrus and diestrus mice. Social dominants and social subordinates in sibling groups did not differ in their exploratory responses to the OF. Different housing procedures, as means to provide different social environment, may differentially induce mild social stress in male and female mice. PMID- 11438370 TI - A model of social stress in dominant mice: effects on sociosexual behaviour. AB - The possibility of socially stressing the dominant/aggressive member of a pair of male mice is tested. Male mice (NMRI outbreed strain) were housed in pairs to assess dominant and subordinate roles by agonistic interactions and urine-marking test. Social stress for dominant males consisted in 30 min/day of exposure to their subordinate partner interacting with a female in the adjacent compartment of the cage, for 9 days. Results showed that dominance status was maintained. Behavioural observations indicated that neither the subordinates nor the dominant males habituated to this experimental procedure. At the end of the chronic stress, dominant animals were given the opportunity to interact for 30 min with a female in their compartment. Results indicated that stressed dominants showed impairment in their sexual behaviour and were more oriented towards the physical environment in comparison with control dominants. The behavioural response to apomorphine (0.25 mg/kg) indicated an alteration of the dopaminergic functioning in socially stressed dominant mice. This study suggests that the characteristics of the stressor and the effects of the chronic social stress could be different, according to male social status. PMID- 11438371 TI - Effect of social defeat in a territorial bird (Parus major) selected for different coping styles. AB - We addressed the questions (i) whether a social defeat triggers similar autonomic and behavioral responses in birds as is known from mammals and (ii) whether individuals that differ in coping style differ in their reaction to a social defeat. Adult captive male great tits (Parus major) from either of two different selection lines for coping style were used to test the effect of social defeat by an aggressive resident male conspecific on subsequent social and nonsocial behaviour, body temperature, breath rate and body mass. These parameters were measured 1 day before (baseline), immediately after and at Days 1 to 3 and 6 after the social interaction took place (Day 0). Social defeat decreased social exploration and increased body temperature substantially for at least 1 day in all birds. Breath rate and body mass were not affected. Birds belonging to the more aggressive and bolder line showed impairment in activity immediately after the social defeat. This is to our knowledge the first report showing that psychosocial stress in birds can have a similar impact as in rodents, but with a shorter recovery time. This might be due to species-specific differences in sensitivity to social stress, or to differences in the way social stress was induced. PMID- 11438372 TI - Social defeat as a stressor in humans. AB - Studies on social defeat in humans, and their similarities with studies on social defeat in animals are reviewed. Studies on social defeat in humans typically are conducted as a branch of social psychology, most often focusing on bullying in schools and in workplaces. Victims of bullying are known to suffer from depression, anxiety, sociophobia, loss of self-esteem, psychosomatic diseases, and other behavioral symptoms. On the other hand, animal studies on social defeat, usually based on the rodent resident--intruder paradigm, present findings related to physiological rather than to behavioral consequences of defeat. The two branches use different terminology, e.g., "dominant" and "subordinate" (animal studies) and "bully" and "victim" (human studies). It is suggested that the two fields could benefit from a mutual exchange in theory and methodology. PMID- 11438373 TI - Gender differences in vulnerability to social stress: a Darwinian perspective. AB - This article offers a theoretical framework based on evolutionary thinking designed to clarify relationships between social stress and mental illness, including the origin of gender differences in vulnerability to stress. From a Darwinian perspective, stress is an interference with evolved behavioral strategies. Human behavior is organized around the pursuit of biological goals, and any social event that interferes with these evolved strategic goals may constitute a stressor. The response to such interference -- the stress response - is made up of physiological, psychological and behavioral components. These components determine how individuals deal with those social events that were likely to reduce inclusive fitness in the ancestral environment. Evolved gender differences in commitment to goals play a role in determining individual differences in response to stressors. When a social stressor interferes with achieving a biological goal, its harmful impact will depend primarily on the importance of the goal to an individual, and the importance assigned to different goals by an individual does not depend exclusively on personal variables and cultural values. Two evolutionary theories are relevant to gender differences in vulnerability to social stress: sexual selection theory and life history theory. Clinical data from patients suffering from depression triggered by social stress are reviewed to test predictions derived from these theories. PMID- 11438374 TI - Residual processing of chromatic signals in the absence of a geniculostriate projection. AB - We have investigated the residual processing of chromatic signals in a subject with unilateral damage to the primary visual cortex using psychophysical, pupillometric and functional magnetic resonance imaging (fMRI) methods. Of particular interest was to establish the correlation between the subject's ability to make use of chromatic signals in the blind hemifield to discriminate between different coloured targets, the corresponding residual pupil colour responses and the level and location of cortical activation generated by the same stimuli as revealed by fMRI. The results obtained using the three different experimental approaches are consistent and suggest that retrograde degeneration of thalamic and retinal chromatic processing mechanisms caused by damage to the primary visual cortex in man does not abolish completely the ability to process chromatic signals particularly when large, long-wavelength stimuli are employed. PMID- 11438375 TI - Where now for the Th1/Th2 paradigm of the gestational uterus? PMID- 11438376 TI - The Th2 cytokines IL-4 and IL-10 are not crucial for the completion of allogeneic pregnancy in mice. AB - The physiological protection from fetal rejection is believed to be dependent on a Th2 type of inflammatory response at the maternal-fetal interface and the cytokines IL-4 and IL-10 have been suggested to play a critical role. We here present data from breeding experiments with IL-10 and IL-4 double-deficient mice indicating that neither maternal nor feto-placental deficiency of these cytokines are crucial for fetal or neonatal survival. The present study does not analyse possible developmental effects of maternal or fetal IL-10 and IL-4 double deficiency in detail, but shows that an apparently normal breeding can be achieved in different crossings, providing that the mice are kept under very clean conditions. PMID- 11438377 TI - Soluble ICAM-1 levels in the serum of endometriotic patients appear to be independent of medical treatment. AB - Adhesion molecules regulate the interaction of cells with the extracellular matrix and/or other cells. The intercellular adhesion molecule-1 (ICAM-1; CD54) is a member of the immunoglobulin superfamily and expressed by several cell types, including leukocytes and endothelial cells. A circulating form of the usually membrane-bound molecule was identified and characterized in normal human serum and in sera from patients with endometriosis. In the present study, we established the serum-soluble ICAM-1 (sICAM-1) levels in patients with endometriosis. We also studied the effect of danazol and leuprorelin acetate depot on the levels of sICAM-1. Thirty-eight women, 18-45 years of age, with regular menses and documented pelvic endometriosis were recruited from a University Hospital setting. Twenty-two women with endometriosis were randomly divided into two groups. Danazol (600 mg) were given every day for 6 months, and 3.75 mg of leuprorelin acetate depot every 28 days for 6 months. Serum sICAM-1 concentrations were measured before, during and after treatment, and its quantitative determination was performed by an ELISA technique using a specific immunoassay. We found that (1) sICAM-1 levels were higher in women with endometriosis in comparison to healthy subjects; (2) the 6 month treatment with danazol or leuprorelin acetate depot increased sICAM-1 levels (P<0.001); (3) 3 months after termination of both treatments, sICAM-1 levels were unchanged. Although the mechanism leading to the increase of sICAM-1 needs to be further clarified, any benefits of medical treatment of endometriosis such as danazol or leuprorelin appear to be independent of changes in ICAM-1 serum levels. PMID- 11438378 TI - Cytokine promoter gene polymorphisms and idiopathic recurrent pregnancy loss. AB - Approximately one in 300 women experience recurrent pregnancy loss (RPL), the aetiology of which is unknown in at least 40% of cases. Previously, some studies have shown increased production of pro-inflammatory cytokines (tumour necrosis factor-alpha and interferon-gamma) and reduced production of anti-inflammatory cytokines (interleukin-10) by circulating blood lymphocytes isolated from these patients when compared with controls. The reasons for this are unclear. The production of these cytokines are partly under genetic control. This study investigated whether polymorphisms in these three cytokine genes known to be associated with either high or low production, are associated with idiopathic RPL. No association was found. It may be that genetic factors are not a major determinant of cytokine production during pregnancy, or alternatively it may be that the observed differences in cytokine production by peripheral lymphocytes do not accurately indicate what is occurring at the local maternofoetal interface during successful and abortive pregnancies. PMID- 11438382 TI - Branching projections of catecholaminergic ventrolateral reticular neurons to the fastigial nucleus and superior colliculus in the rat: triple labelling procedure. AB - In this study, we employed triple fluorescent-labelling to reveal the distribution of the catecholaminergic neurons within rostral ventrolateral reticular nucleus which supply branching collateral input to the superior colliculus (SC) and to the cerebellar fastigial nucleus (FN). The catecholaminergic identity of the neurons was revealed by immunocytochemical detection of the biosynthetic enzyme, tyrosine hydroxylase. The projections were defined by injections of two retrograde tracers: rhodamine and fluoro gold in the SC and FN, respectively. PMID- 11438383 TI - An unusual population of pyramidal neurons in the anterior cingulate cortex of hominids contains the calcium-binding protein calretinin. AB - In the context of an on-going comparative analysis of primate neocortex evolution, we describe the occurrence and distribution of a previously unrecognized group of pyramidal neurons, restricted to the superficial part of layer V in the anterior cingulate cortex of hominids and characterized by immunoreactivity to the calcium-binding protein, calretinin. These neurons were rare in orangutans, more numerous in gorillas and common chimpanzees, while humans had the highest numbers. These calretinin-containing pyramidal cells were not observed in the cingulate cortex of any other primate or mammalian species. This finding, together with other recent observations on the hominoid cingulate cortex, is interesting when considering primate neocortical evolution, as it indicates possible adaptive and anatomical modifications in a cortical region critical for the integration of many aspects of autonomic function, vocalization, and cognitive processes. PMID- 11438384 TI - Distribution of median, ulnar and radial motoneurons in the monkey spinal cord: a retrograde triple-labeling study. AB - Topographic distribution of motoneurons innervating hand muscles through the median (Mn), ulnar (Ul), or radial (Rd) nerves was examined using a retrograde multiple-labeling technique in the macaque monkey. The Mn and Ul motoneurons, i.e. flexor motoneurons, were distributed from C6 to T2 and from C7 to T2 segments of the spinal cord, respectively, while the Rd motoneurons, i.e. extensor motoneurons, were distributed from C4 to T2. The present study further revealed partial intermingling of the cell bodies and partial overlap of the dendritic fields among the motoneurons projecting through different nerves, indicating that subregions of motoneuronal pool participate in coordination between the flexor and extensor, or among the flexor muscles. It was suggested that there exists a control mechanism for precise hand movements in the spinal cord. PMID- 11438385 TI - Menstrual cycle effects on postural stability but not optokinetic function. AB - We investigated the effects of the menstrual cycle on visual-vestibular interaction by measuring optokinetic function and postural stability at different phases of the cycle. Menstrual cycle phase had no significant effect on gaze holding, optokinetic nystagmus slow phase velocity, amplitude or frequency, circularvection latency or optokinetic afternystagmus frequency, amplitude or duration. While menstrual cycle phase had no significant effect on anterior posterior sway, it did significantly affect lateral sway (P<0.001), with sway on day 5 significantly greater than on days 12 and 21 (P<0.05 and 0.01, respectively), and sway on day 25 significantly greater than that on day 21 (P<0.05). PMID- 11438386 TI - Excitotoxic preconditioning elicited by both glutamate and hypoxia and abolished by lactate transport inhibition in rat hippocampal slices. AB - Ischemic preconditioning (PC) of heart and brain is a well-documented phenomenon. However, the mechanism underlying the increased resistance to severe ischemia by a preceding mild ischemic exposure remains unclear. Over a decade ago, we demonstrated the existence of hypoxic PC in the hippocampal slice preparation. Here we report the ability of a short exposure to toxic levels of glutamate to heighten the tolerance of hippocampal slices to a subsequent, longer exposure to the excitotoxin. Glutamate PC could also be induced by a short hypoxic exposure, suggesting a common mechanistic pathway for all PC stimuli. Since glutamate receptor activation and hypoxia increase tissue lactate production, a-cyano-4 hydroxycinnamate was applied during the PC period to completely abolished PC. These results indicate that excitotoxic PC and hypoxic PC share similar mechanisms that possibly involve lactate production and its neuronal utilization. PMID- 11438387 TI - Estrogen decreases the responsiveness of subfornical organ neurons projecting to the hypothalamic paraventricular nucleus to angiotensin II in female rats. AB - Extracellular single-unit activity was recorded from subfornical organ (SFO) neurons antidromically identified as projecting to the hypothalamic paraventricular nucleus (PVN) in urethane-anesthetized ovariectomized female rats that were treated with either propylene glycol (PG) vehicle or estradiol benzoate (EB). No significant differences were observed between the PG- and EB-treated rats in the latency, conduction velocity, or threshold of antidromic activation. The mean spontaneous firing rate was significantly lower and the refractory period was significantly longer in the EB-treated rats. In the identified units that were activated by angiotensin II (ANG II) applied iontophoretically, the amount of excitatory response to intracarotid administration of ANG II was much greater in the PG-treated than in the EB-treated rats. These results suggest that estrogen may decrease the responsiveness of SFO neurons projecting to the PVN to circulating ANG II. PMID- 11438388 TI - Spatial performance correlates with long-term potentiation of the dentate gyrus but not of the CA1 region in rats with fimbria-fornix lesions. AB - Although hippocampal long-term potentiation (LTP) is generally assumed to be a cellular mechanism of learning and memory, there has not been definitive evidence for this hypothesis. In the present study, therefore, we addressed the possible relationship between spatial learning ability and LTP by using rats with bilateral fimbria-fornix lesions. The animals were tested for spatial performance in spontaneous alternation behaviors with further in vivo investigation of LTP. The behavioral parameters of spatial memory showed a significant correlation with LTP in the dentate gyrus, but we found no evidence for a linkage with LTP in the CA1 region. Thus, LTP in the dentate gyrus may be important for spatial cognitive ability. PMID- 11438389 TI - DL-Tetrahydropalmatine may act through inhibition of amygdaloid release of dopamine to inhibit an epileptic attack in rats. AB - DL-Tetrahydropalmatine (THP), an active component isolated from corydalis (a Chinese herbal medicine), possesses analgesic effects. Systemic administration of picrotoxin (3-4 mg/kg) produced increases of locomotion (including horizontal motion, vertical motion, and total distance traveled), elevations of turnings (including both clockwise and anticlockwise), and inhibition of postural freezing in freely moving rats, and increases of amygdaloidal release of dopamine in anesthetizes rats. All the afore-mentioned activity measures induced by picrotoxin were suppressed following THP pretreatment. The results indicate that THP may act through inhibition of amygdaloid dopamine release to inhibit an epileptic attack. PMID- 11438390 TI - Vasoactive intestinal peptide and related molecules induce nitrite accumulation in the extracellular milieu of rat cerebral cortical cultures. AB - Nanomolar concentrations of vasoactive intestinal peptide (VIP), picomolar concentrations of stearyl-norleucine17-VIP (SNV) and femtomolar concentrations of NAPVSIPQ (NAP), an 8-amino-acid peptide derived from the VIP-responsive activity dependent neuroprotective protein, provide broad neuroprotection. In rat cerebral cortical cultures, 10(-16)-10(-7) M NAP increased intracellular cyclic guanosine monophosphate (cGMP) (2.5-4-fold) and 10(-10) M NAP increased extracellular nitric oxide (NO) by 60%. In the same culture system, VIP and SNV (at micromolar concentrations) increased extracellular NO by 45-55%. The NAP dose required for cGMP increases correlated with the dose providing neuroprotection. However, the concentrations of NAP, SNV and VIP affecting NO production did not match the neuro-protective doses. Thus, NO may mediate part of the cell-cell interaction and natural maintenance activity of VIP/SNV/NAP, while cGMP may mediate neuroprotection. PMID- 11438391 TI - Ability of rat microglia to uptake extracellular glutamate. AB - Since it has been suggested that microglia in vivo act as glutamate scavengers, this possibility was investigated in primary cultured microglia. The microglia showed specific abilities to uptake (14)C-glutamate depending on incubation time and numbers of cells used. The activity was suppressed by a specific inhibitor for a glial cell-type transporter, glutamate transporter-1 (GLT-1) (EAAT2). However, that of cultured astrocytes was not affected. These results suggest that microglia uptake glutamate by means of GLT-1. Supporting these results, immunoblotting revealed the presence of GLT-1 in the microglia, while only glutamate-aspartate transporter (GLAST) (EAAT1: another glial cell-type transporter) was detected in the astrocytes. All together, these results indicate that microglia can act as glutamate scavengers in vivo by expressing the glutamate transporter GLT-1. PMID- 11438392 TI - Dopaminergic inhibition of excitatory inputs onto pyramidal tract neurons in cat motor cortex. AB - The role of dopamine (DA) on motor cortical pyramidal tract neurons (PTNs) was studied in anesthetized cats with in vivo extracellular recordings in response to transcallosal (TC) and ventrolateral (VL) thalamic stimulations. An antidromic PT potential was evoked to recognize PTNs. In most PTNs, iontophoretic application of DA significantly reduced the spike activity exerted by 20 single-pulse stimulations. Both D(1)-like and D(2)-like receptor antagonists blocked (disinhibited) the effect in a similar way regardless of TC and VL stimulations, suggesting colocalization of two receptors. Except for the presence of jittering, the mean latency was usually fixed and short. These findings indicate that ventral midbrain DA imposes an intense suppression in modulating PTNs response to both callosal and thalamocortical excitatory inputs in motor cortex. Such DAergic suppression could play pivotal role to improve motor and sensorimotor signal integration. PMID- 11438393 TI - Anorectic effect of leptin is mediated by hypothalamic corticotropin-releasing hormone, but not by urocortin, in rats. AB - Leptin is a key afferent signal that decreases food intake and increases energy expenditure by acting on the specific receptors in the hypothalamus. Corticotropin-releasing hormone (CRH) and its homologous peptide, urocortin, are also known to have a potent anorectic effect when given intracranially. To determine possible involvement of these two peptides in the leptin-induced anorexia, in the present study, food intake was measured in rats pretreated with antibodies against CRH and urocortin. In the control rats without antibody pretreatment, intracerebroventricular (icv) injection of leptin (0.1-1 microg/rat) suppressed nocturnal food intake. The anorectic effect of leptin was substantially attenuated in rats pretreated with icv injection of an anti-CRH antibody, but not with an anti-urocortin antibody. These results suggest that the anorectic effect of leptin is mediated by CRH, but not by urocortin, in the hypothalamus. PMID- 11438394 TI - Changes of the expression of 5-HT receptor subtype mRNAs in rat dorsal root ganglion by complete Freund's adjuvant-induced inflammation. AB - By using the reverse transcriptase polymerase chain reaction technique, the expression of 5-hydroxytryptamine (5-HT) receptor subtype mRNAs in the rat lumbar dorsal root ganglion (DRG) was investigated following unilateral injection of complete Freund's adjuvant (CFA) into the rat hind paw. The results showed that 5 HT(1A), 5-HT(1B), 5-HT(1D), 5-HT(1F), 5-HT(2A), 5-HT(3), 5-HT(4), 5-HT(5A) and 5 HT(7) receptor subtypes were present in the rat lumbar DRG. CFA injection resulted in a significant increase in mRNA level of 5-HT(1A), 5-HT(1B), 5-HT(1F), 5-HT(2A), 5-HT(3), 5-HT(4) and 5-HT(7) receptor subtypes and a marked induction of 5-HT(2C) subtype mRNA in the DRG. The present results suggest the important roles for these 5-HT receptor subtypes in generating peripheral nociceptive signaling and provide evidence to elucidate the mechanism of 5-HT in nociception. PMID- 11438395 TI - Expression of c-fos mRNA is increased and related to dynorphin mRNA expression following excitotoxic spinal cord injury in the rat. AB - Previous studies have demonstrated that excitotoxic spinal cord injury (SCI) created by the intraspinal injection of quisqualic acid (QUIS) is capable of inducing opioid peptide gene expression within the spinal cord and cortex. The opioids are classically involved in the suppression of pain transmission but specifically, dynorphin, has been implicated in the secondary pathophysiologic response to SCI. Activation of the immediate early gene, c-fos, has been implicated in the induction of preprodynorphin (PPD) gene expression and therefore, may be an important intermediate step in the generation of the opioid response to SCI. The purpose of this study was to investigate whether intraspinal QUIS injection induces c-fos expression within the spinal cord. Male, Long-Evans, adult rats (n=5) received an intraspinal injection of 1.2 microl of 125 mM QUIS directed at spinal segments T12-L2. Four hours post-injection brain and spinal cord tissues were removed and processed for in situ hybridization. Integrated density of c-fos and PPD mRNA expression was increased in the spinal dorsal horn following QUIS injection as compared to sham-injected animals. This indicates that SCI rapidly induces c-fos and PPD expression and suggests that c-fos plays a role in the induction of PPD expression. PMID- 11438396 TI - In vivo 3D visualization of normal pyramidal tracts in human subjects using diffusion weighted magnetic resonance imaging and a neuronavigation system. AB - We describe the potential of anisotropic diffusion weighted imaging to visualize the course of large cerebral fiber tracts. Five healthy volunteers were investigated at a field strength of 1.5 Tesla, employing a spin-echo diffusion weighted sequence with gradient sensitivity in six non-collinear directions to visualize the course of the pyramidal tracts. The pyramidal tracts were segmented and reconstructed for three-dimensional visualization. Reconstruction results together with a fusioned high resolution 3D T1 weighted image data set were available in a customized neuronavigation system. Origination in the primary motor cortex, convergence in the centrum semiovale, the posterior limb of the internal capsule, the cerebral peduncles, the splitting at the level of the pons, and the pyramidal decussation were identified in all subjects. Fiber tract maps might have the prospect of guiding neurosurgical interventions, especially when being linked to a neuronavigation system. Other potential applications include the demonstration of the anatomical substrate of functional connectivity in the human brain. PMID- 11438397 TI - Changes in cortical noradrenergic axon terminals of locus coeruleus neurons in aged F344 rats. AB - The noradrenergic innervations and noradrenaline contents of the frontal cortex in two age groups (9 and 25 months) of male F344 rats have been quantified by electrophysiological and biochemical methods. In the electrophysiological study, the percentage of locus coeruleus (LC) neurons activated antidromically from the frontal cortex decreased with age. In contrast, the percentage of LC neurons showing multiple antidromic latencies, which suggests axonal branching of individual LC neurons, increased markedly between 9 and 25 months in the frontal cortex. In the biochemical study, we found no significant difference in noradrenaline levels in the cortical terminal fields of LC neurons during aging. These results suggest that LC neurons give rise to axonal branches to retain noradrenaline levels in their target fields in the aged brain. Our findings show that LC neurons preserve a strong capability for remodeling their axon terminals even in the aged brain. PMID- 11438398 TI - Fluorometric detection of platelet activating factor receptor in cultured oviductal epithelial and stromal cells and endometrial stromal cells from bovine at different stages of the oestrous cycle and early pregnancy. AB - During the oestrous cycle and early pregnancy, the oviduct and uterus undergo a variety of morphological and physiological modifications in which the platelet activating factor receptor (PAF-R) plays an important role. PAF-R levels were quantified in bovine oviductal epithelial and stromal cells and endometrial stromal cells at days 2 to 4, 12, and 20 of the estrous cycle and during early pregnancy. Cells were grown in vitro and their intracellular PAF-R concentration was measured by flow cytometry using a polyclonal anti-PAF-R antibody system. A significant increase (P < 0.05) in the portion of PAF-R-positive oviductal epithelial and stromal cells was detected in both non-pregnant and pregnant cattle on days 2 to 4 in comparison to day 12 and 20. In endometrial stromal cells derived from day 20 pregnant bovine, a significant increase (P < 0.05) in PAF-R staining was observed in comparison to the day 20 non-pregnant and days 2 to 4 or 12 pregnant and non-pregnant animals. The PAF-R was detected in oviductal cells by using immunoblotting and immuno-gold postembedding method. Positive binding of the anti-PAF-R antibody was found on the cell membrane and in the cytoplasm. We concluded that the increased PAF-R concentration measured in cultured oviductal epithelial and stromal cells of cyclic and pregnant heifers on days 2 to 4 was hormonally regulated. The increased PAF-R in endometrial stromal cells on day 20 of pregnant heifers was a pregnancy-specific effect and may mediate a local increase in endometrial vascular permeability known to precede the implantation. PMID- 11438399 TI - Selection for growth rate or against back fat thickness in pigs is associated with changes in growth hormone axis plasma protein concentration and mRNA level. AB - Selection for increased growth rate or decreased back fat thickness results in concomitant changes in endocrine and metabolic status. Growth hormone (GH) changes in blood plasma concentration related to selection for growth rate and fat deposition were reported in pigs. The molecular mechanisms regulating selection-induced changes in GH plasma concentration remain largely unknown. We investigated selection-associated changes in GH axis parameters in 2 pig lines selected for increased growth rate (F-line), or decreased back fat thickness (L line), respectively. First, we investigated selection-associated changes in GH pulse parameters. In both selection lines we found each generation a declining GH peak maximum concentration and area under the GH curve. GH pulse width was not associated with generation number. In both lines generation number was associated with a declined pulse interval, indicating that the number of pulses per day increased on average with 1 pulse per 24 h per generation. Second, plasma concentration of GH axis related Insulin-like growth factor-I (IGF-I) and insulin were investigated. Plasma IGF-I concentration was not associated with generation number in the F-line. Mean plasma insulin concentration declined each generation in both lines. Third, we investigated changes in GH and Pit-1 mRNA levels. In both selection lines GH and Pit-1 mRNA levels increased approximately 50% each generation. The high SD of the GH mRNA levels in both lines may suggest that the GH mRNA levels are pulsatile in vivo. We postulate a molecular mechanism that may explain how selection is associated with increased GH mRNA levels and GH pulse numbers, while lowering GH release per pulse. PMID- 11438400 TI - Characterization of the feline thyroglobulin promoter. AB - The feline thyroglobulin promoter was identified by a combination of standard polymerase chain reaction (PCR) techniques, using primers designed according to regions of homology in published sequences from other species, then adaptor ligated PCR. A 310 bp fragment of the feline thyroglobulin promoter was generated, including 8 nucleotides of adaptor sequence at the 5' end and, based on the putative transcription start site, 36 nucleotides of the thyroglobulin mRNA (untranslated portion). The homology between the feline promoter sequence (from 193 bp upstream to the putative cap site) and canine, bovine and human sequences was 89%, 81% and 78%, respectively. Transient transfection studies, using reporter constructs in which the feline promoter controlled expression of chloramphenicol acetyl transferase, demonstrated promoter activity in thyroid cells, but no activity in non-thyroid cells. The data presented here demonstrate that the feline thyroglobulin promoter may provide a targeting mechanism for somatic gene therapy of feline thyroid disease. PMID- 11438401 TI - Expression of insulin-like growth factor binding protein (IGFBP)-3, and the effects of IGFBP-2 and -3 in the bovine corpus luteum. AB - The present study was conducted to gain insight into the insulin-like growth factor (IGF) system in the bovine corpus luteum (CL). Specific aims were to measure the levels of IGF binding protein-3 (IGFBP-3) and RNA encoding IGFBP-3 in the CL throughout diestrus, and to investigate the effects of IGFBP-2 and -3 on IGF-I-stimulated progesterone (P4) production and IGF-I-receptor binding. Bovine CL were collected from a local abattoir and classified according to stage of diestrus based on anatomical characteristics. Corpora lutea from early, mid and late diestrus were each analyzed for the presence of IGFBP-3 by ligand blot analysis, and for RNA encoding IGFBP-3 by Northern blot analysis. Dissociated cells from mid-cycle CL were treated with IGF-I, IGFBP-2 or -3, or a combination of IGF-I and IGFBP-2 or -3. The effect of IGFBP-2 and IGFBP-3 on [(125)I] IGF-I binding to its receptor on CL plasma membranes also was investigated. IGFBP-3 protein and RNA expression were higher in early CL, compared to mid or late CL (p < 0.05). IGF-I stimulated P4 production in a dose-dependant manner (p < 0.05). IGFBP-2 and -3 blocked the stimulatory effect of IGF-I on P4 production (p < 0.05). Both IGFBP-2 and -3 inhibited [(125)I]-IGF-I binding to its receptor in a dose-dependant manner. These results demonstrate that IGFBP-3 protein and RNA are expressed predominantly during early diestrus in the bovine CL. Moreover, both IGFBP-2 and -3 can modulate IGF-I actions in the CL by interfering with binding of IGF-I to its receptor. PMID- 11438402 TI - Hormone control of prostaglandin F(2 alpha) production and oxytocin receptor concentrations in bovine endometrium in explant culture. AB - The effect of progesterone and estradiol on basal and oxytocin-stimulated prostaglandin F(2 alpha) production and on oxytocin receptor concentrations in endometrium from long term ovariectomized cows was investigated using an explant culture system. Uteri were obtained from cows at slaughter and endometrial explants were cultured in triplicate for up to 96 h in either control media, or media containing progesterone or estradiol. Basal prostaglandin F(2 alpha) production was unaffected by progesterone treatment but was stimulated by estradiol treatment in a dose dependent manner. Oxytocin receptor concentrations remained unchanged in control culture and were unaffected by treatment with estradiol while treatment with progesterone caused a dose-dependent inhibition. Responsiveness to oxytocin, in terms of increased prostaglandin F(2 alpha) production, developed "spontaneously" over the first 24 h of culture and was unaffected by treatment with progesterone or estradiol. In summary the results reveal a dose-dependent inhibition of oxytocin receptor concentration by progesterone and a dose-dependent stimulation of basal PGF(2 alpha) release by estradiol. The reason for the "spontaneous" development of responsiveness to oxytocin remains unknown but may result from the removal of tissue from the influence of an, as yet unidentified, inhibitory factor. PMID- 11438403 TI - Aldosteronoma in a dog with polyuria as the leading symptom. AB - In a 10-year-old castrated male shorthaired German pointer polyuria was associated with slight hypokalemia, hypophosphatemia and alkalosis, as well as elevated plasma concentrations of a glucocorticoid-inducible iso-enzyme of alkaline phosphatase. Repeated measurements of urinary corticoids and normal suppressibility of the hypothalamus-pituitary-adrenocorticial axis excluded glucocorticoid excess. Urine osmolality (Uosm) did not increase during administration of the vasopressin analogue desmopressin. At the time water deprivation had caused Uosm to rise from 300 to 788 mOsm/kg, there was also plasma hypertonicity. During hypertonic saline infusion the osmotic threshold for vasopressin release was increased. The combination of elevated plasma aldosterone concentrations and unmeasurably low plasma renin activity pointed to primary hyperaldosteronism. As initially computed tomography (CT) did not reveal an adrenocortical lesion, the dog was treated with the aldosterone antagonist spironolactone. This caused Uosm to rise in a dose-dependent manner. However, well-concentrated urine was only achieved with doses that gave rise to adverse effects. Once repeated CT, using 2-mm-thick slices, had revealed a small nodule in the cranial pole of the left adrenal, unilateral adrenalectomy was performed which resolved the polyuria completely. Also the plasma concentrations of kalium, aldosterone and renin activity returned to within their respective reference ranges. The adrenocortical nodule had the histological characteristics of an aldosteronoma, with the non-affected zona glomerulosa being atrophic.In this dog with primary hyperaldosteronism the polyuria was characterized by vasopressin resistance and increased osmotic threshold of vasopressin release, similar to the polyuria of glucocorticoid excess. The possibility is discussed that the polyuria of glucocorticoid excess is actually a mineralocorticoid effect. PMID- 11438404 TI - The art of quality assessment of RCTs included in systematic reviews. AB - The best evidence on the efficacy of medical interventions is provided by high quality trials summarized in high-quality systematic reviews or meta-analyses. The methodological quality of studies included in a systematic review can have a substantial impact on the estimates of the treatment effect and therefore on the conclusions of such a review. But what is the empirical evidence to support quality assessment of randomized clinical trials (RCTs)? We elaborate on questions such as: what is the concept of quality of individual studies (RCTs), can quality be measured validly and reliably? Plans for future research on this issue are proposed. PMID- 11438405 TI - Characteristics of meta-analyses related to acceptance for publication in a medical journal. AB - Editors of medical journals select manuscripts for publication based, in part, on the perceived quality of the manuscript submitted. The objective of this study was to describe associations between acceptance for publication and quality related methodologic characteristics of meta-analyses. This was a prospective observational study. The setting was editorial offices of JAMA and offices of external reviewers. The manuscripts reviewed were 112 consecutive meta-analyses submitted to JAMA during 1996 and 1997 whose authors agreed to participate. The main outcome measures were ratings of 16 methodologic characteristics reflecting quality of the meta-analysis and acceptance for publication. A "high" rating for one methodologic characteristic, whether the report of the meta-analysis provided sufficient detail to enable replication, was related significantly to publication (RR = 2.79, 95% CI = 1.13-6.89). This relationship persisted when other variables were controlled for in the model. Generally, rejected manuscripts had fewer factors rated "high," but differences were not significant. We found that inclusion of sufficient detail to allow a reader to replicate meta-analytic methods was the only characteristic related to acceptance for publication. PMID- 11438406 TI - Causes and consequences of comorbidity: a review. AB - A literature search was carried out to identify and summarize the existing information on causes and consequences of comorbidity of chronic somatic diseases. A selection of 82 articles met our inclusion criteria. Very little work has been done on the causes of comorbidity. On the other hand, much work has been done on consequences of comorbidity, although comorbidity is seldom the main subject of study. We found comorbidity in general to be associated with mortality, quality of life, and health care. The consequences of specific disease combinations, however, depended on many factors. We recommend more etiological studies on shared risk factors, especially for those comorbidities that occur at a higher rate than expected. New insights in this field can lead to better prevention strategies. Health care workers need to take comorbid diseases into account in monitoring and treating patients. Future studies on consequences of comorbidity should investigate specific disease combinations. PMID- 11438407 TI - Problems in determining occurrence rates of multimorbidity. AB - This article describes methodological decisions that have to be made when studying multiple pathology and presents appropriate analytical techniques. The main question of this article is: how can comorbidity and multimorbidity be operationalized with respect to the number and type of diseases studied, and which analytic approaches are available for the evaluation of multiple pathology? Choices regarding the number and type of diseases studied have great impact on the observed incidence and prevalence rates of comorbidity and multimorbidity. These rates are largely dependent on age, sex, and other determinants. In addition to crude descriptive measures, odds ratios and relative risks can be used to study comorbidity, whereas multimorbidity can be studied using observed/expected ratios. While basic analyses of comorbidity can be performed using standard statistical packages, two additional programs were developed for the analysis of the distribution of multimorbidity and statistically unexpected comorbidity, respectively. As some analyses are addressing multicomparisons, external validity testing is recommended. PMID- 11438408 TI - Prevalence of morbidity and multimorbidity in elderly male populations and their impact on 10-year all-cause mortality: The FINE study (Finland, Italy, Netherlands, Elderly). AB - Older males are known to carry, more likely than younger people, one or more chronic diseases with an expected impact on mortality. This study was aimed at identifying the relationship of prevalent chronic diseases in elderly populations of different countries with all-cause mortality. Men aged 65-84 from defined areas were enrolled in Finland (N=716), the Netherlands (N=887) and Italy (N=682). They were survivors of cohorts studied for 25 years within the Seven Countries Study. Major chronic diseases were diagnosed at entry. Ten-year follow up for mortality was completed. Entry prevalence of selected chronic diseases was higher in Finland (56%) than in Italy (51%) and the Netherlands (44%). Ten-year age-adjusted death rates from all causes were higher in Finland (565 per 1000) and lower in the Netherlands (478 per 1000) and Italy (445 per 1000). The absolute risk of death related to chronic disease was high in the three countries, but was higher in Finland than in the Netherlands and Italy. The most lethal condition was stroke, with 10-year death rates of 806 per 1000 in Finland and 707 and 729 per 1000 in the Netherlands and Italy, respectively. The relative risk of all-cause mortality for a set of seven chronic diseases (coronary heart disease, heart failure, claudicatio intermittens, cerebrovascular accidents, diabetes, COPD and cancer) adjusted by age, other diseases and cohort was less than two for each condition, except cerebrovascular accidents in the Netherlands (RR 2.20). In general, relative risk was higher in Finland, intermediate in the Netherlands and lower in Italy, where only cerebrovascular accidents, intermittent claudication, diabetes and the presence of any chronic condition had a significant relative risk. About one third of men had one chronic disease, and between 10% and 15% had two diseases. The coexistence of any two or three chronic conditions was associated with a relative risk of 2 or more in Finland and the Netherlands and less than 2 in Italy. In these elderly men prevalent morbidity and comorbidity was relatively common and it explained a large proportion of excess in all-cause mortality in 10 years of follow-up. PMID- 11438409 TI - Accuracy of administrative data to assess comorbidity in patients with heart disease. an Australian perspective. AB - The objective of this study was to determine the accuracy of administrative data (by use of hospital discharge codes) for measuring comorbidity in patients with heart disease. One thousand seven hundred and sixty-five medical records of subjects admitted to hospital for AMI, unstable angina, angina pectoris, chronic IHD or heart failure were reviewed. The number and types of comorbidities were determined from the medical records (regarded as the "gold standard"). These were compared with the 10 discharge codes obtained from the hospital administrative records (referred to as the "administrative data"). The rate of false-negative and false-positive comorbidity diagnoses were determined. Twenty of the 21 comorbidities studied were underreported in the administrative data. For these 20 comorbidities, the median false-negative rate was 49.5% and ranged from 11% for diabetes to 100% for dementia. False-positive rates were low, less than 1.5%, except for chronic arrythmia (4.8%) and hypertension (4.2%). Mean percent agreement was high, ranging from 88% for hypertension to 100% for AIDS/HIV. Administrative data based on hospital discharge codes consistently underestimate the presence of comorbid conditions in our population. This has implications for administrators when estimating mortality, length of stay and disability. Researchers also need to be aware when using administrative data based on hospital discharge codes to assess subject's comorbidities that they may be widely underreported. PMID- 11438410 TI - Risk adjustment for older hospitalized persons: a comparison of two methods of data collection for the Charlson index. AB - To compare Charlson indices based on chart data and ICD-9 data for agreement overall and on rating specific comorbid conditions, and to compare mortality risks associated with these indices. Prospective cohort study. Six general medicine wards at Yale-New Haven Hospital. 524 consecutive patients who had no clinical evidence of delirium at enrollment, admitted between November 6, 1989 and July 31, 1991, aged 70 years or older. Death within 1 year of the index hospital admission date. Scores using the chart-based data were significantly higher than those using ICD-9 data. About half of the individual conditions showed fair-to-good agreement between the two scores, whereas the other half showed poor agreement. A comparison of mortality prediction indicated that the weightings assigned to individual comorbidities differed substantially from those used in Charlson's original index. While mortality prediction of each individual index was comparable, the ICD-9 and chart indices contributed independently to mortality prediction in the presence of the other. Low agreement between Charlson scores based on the two methods of data collection and their cumulative contribution to mortality prediction suggest that these indices may include different information. Our results suggest that the original Charlson index may not provide optimal risk adjustment for elderly general medicine samples. We suggest development of an empirically-derived index of comorbid conditions and weights may be warranted for older general medical patients. PMID- 11438411 TI - Risk factors for neuroblastoma at different stages of disease. Results from a population-based case-control study in Germany. AB - Neuroblastoma is one of the childhood cancers included in two recent population based case-control studies in West Germany. Altogether, 183 children under the age of 8 with neuroblastoma diagnosed in 1988-1994 and 1785 control children sampled from population registration files participated. Information on potential risk factors was obtained from the children's parents by a self-administered questionnaire and subsequent telephone interview. We observed positive associations with the use of oral contraceptives or other sex hormones during pregnancy (particularly with male offspring), a shorter gestational duration, lower birth weight, and maternal alcohol consumption during pregnancy. While the association with maternal use of oral contraceptives or sex hormones was strong for stages I/II (odds ratio 4.5, 95% confidence interval 1.2-16.5), the associations with shorter gestation duration (odds ratio 3.4, 95% confidence interval 1.7-6.7) as well as maternal alcohol consumption during pregnancy (>7 glasses/week odds ratio 5.2, 95% confidence interval 1.3-20.6) were observed only for the unfavourable advanced stages. It is notable that the associations in our study were either observed only for the advanced stages of disease or only for the less advanced stages, but not for both subgroups. This adds to evidence for the hypothesis that neuroblastoma consists of at least two distinct disease entities, which differ in clinical stage at the time of diagnosis. PMID- 11438412 TI - Urinary tract infection among women aged 40 to 65: behavioral and sexual risk factors. AB - We conducted a case-control study to explore the role of health behavior and sexual and medical history on urinary tract infection (UTI) risk among otherwise healthy women aged 40-65. Cases and controls were recruited from nine practices and clinics in Michigan and a single clinic in Israel. In both countries, several factors were reported significantly more frequently among UTI cases than controls: a previous UTI within 12 months, incontinence symptoms, a recent episode of 30-plus minutes of cold hands, feet, back or buttocks, and recent antibiotic use. Cases were less likely than controls to report recent estrogen use, but the results were only statistically significant in Michigan. Sexual activity during the previous 2 weeks and having ceased menses were modestly, but not statistically significantly, protective at both study sites. Risk factors for UTI among women 40-65 differ from those for younger women and these differences cannot be attributed solely to changes in menopausal status. PMID- 11438413 TI - Socio-demographic factors, health behavior and late-stage diagnosis of breast cancer in Germany: a population-based study. AB - Late-stage diagnosis of breast cancer is associated with poor survival. Identification of individuals at high risk of late-stage diagnosis could be an effective step to reduce breast cancer mortality. We examined the association of socio-demographic factors and health behavior with breast cancer stage in a population-based sample of 380 female breast cancer patients in Saarland, Germany. Overall, 182 women (47.9%) were diagnosed with late-stage (regional or distant) breast cancer. After control for potential confounding by multivariate logistic regression, an increased risk of late-stage diagnosis was observed for older age (OR = 1.8; 95% CI 1.0-3.2), foreign nationality (OR = 3.9; 95% CI 0.7 20.8), living in large households (OR = 1.7; 95% CI 1.0-2.9), non-participation in general health check-up (OR = 1.5; 95% CI 0.9-2.4) and low interest in health care (OR = 1.6; 95% CI 1.0-2.7). The proportion of late-stage cancer was clearly decreased when tumors were detected by screening (OR = 0.4; 95% CI 0.2-0.8). Certain socio-demographic factors and characteristics of health behavior seem to represent independent risk indicators of late-stage diagnosis. PMID- 11438414 TI - Antidepressant use and the risk of breast cancer: a non-association. AB - OBJECTIVE: Animal studies have suggested that some antidepressant medications may act as breast tumor promoters and recent epidemiologic studies of antidepressant use and breast cancer outcomes in humans have also reported such associations. DESIGN: Retrospective cohort study. STUDY POPULATION: 38,273 women who filled a prescription for any of a number of antidepressants and 32,949 who filled a prescription for any other medication during 1989-1991; all were > or =20, enrolled in Medicare, Medicaid or the Pharmaceutical Assistance to the Aged and Disabled (PAAD) programs of New Jersey, and free of evidence of breast cancer. DATA COLLECTION: Antidepressant use was assessed over a period lasting up to 24 months. Subjects were followed for a maximum of 7.5 years; those who had a first diagnosis of breast cancer in the New Jersey Cancer Registry at least 3 months after their index date were considered incident breast cancer cases. Other covariates, including demographic, clinical, and health care utilization variables were also assessed. MAIN OUTCOME OF INTEREST: Adjusted hazard ratio (HR) of developing breast cancer, based on multivariable proportional hazards models. RESULTS: Use of antidepressants was unrelated to the development of breast cancer (adjusted HR = 1.04; 95% CI 0.87-1.25). No elevated risks were found for specific antidepressants, including agents found to be breast tumor promoters in animal studies, as well as drugs thought to be associated with breast cancer in prior epidemiologic studies. There was no suggestion that breast cancer risks were increased with more intensive use of antidepressants or that antidepressant use is associated with a more severe stage of cancer at diagnosis. CONCLUSIONS: Despite recent reports from much smaller epidemiologic studies or laboratory animals, these results provide reassurance that antidepressant use is not associated with the development of breast cancer. PMID- 11438415 TI - Pulse pressure and heart rate: independent risk factors for cancer? AB - In the present study, the roles of heart rate (HR) and pulse pressure (PP) on cancer mortality, after taking into account physical activity, cigarette smoking, alcohol consumption and other confounding factors or underlying disease, were examined in men. The study included 125,513 men aged 20 to 95 years who had a health check-up at the IPC Center between 1978 and 1988. HR and PP were classified into three groups: < 60, 60-80, > 80 bpm for HR and < 50, 51-64, > or = 65 mmHg for PP. Adjusted risk ratios related to the increment from one class of HR or PP to the next for all cancer mortality were 1.4 (1.2-1.5) and 1.3 (1.1 1.4), respectively. This relationship was independent of several known risk and confounding factors, especially cigarette smoking and physical activity, and could not be explained by the presence of underlying disease. PMID- 11438416 TI - Sensitivity of self-reports of cancer in a population-based prospective study: JPHC Study Cohort I. AB - The sensitivity of self-reports of cancer may differ among cultures, and not many studies have dealt with it. The authors measured the sensitivity of 615 individuals from the JPHC Study Cohort I in Japan who responded to a questionnaire in 1995 and had had a cancer registry report since 1990. Sensitivity as calculated using registered cancer as the gold standard was 0.36 for any cancer, but varied considerably by site: 0.81, 0.42, 0.41, 0.26, 0.14 and 0.08 for breast, uterus, stomach, lung, colorectum and liver, respectively. This sensitivity by site relates positively with the cancer incidence/death ratio (an index of prognosis) of each site. A false name was reported in 71% of colorectal cancer cases (namely polyp) and in 51% of stomach cases (ulcer and/or polyp). In conclusion, the sensitivity of self-reports of cancer was much lower in this cohort than in the US results except for breast cancer. PMID- 11438417 TI - Evaluating treatment strategies in chronic lymphocytic leukemia: use of quality adjusted survival analysis. AB - To assess comparatively, in terms of quality-adjusted survival, three front-line treatments in patients with stage B- or C-chronic lymphocytic leukemia (CLL). To describe better and compare the survival after randomization of patients from the CLL90 trial that randomly compared ChOP (cyclophosphamide, doxorubicin, oncovin, prednisone), CAP (cyclophosphamide, doxorubicin, prednisone) and fludarabine in advanced CLL, we performed a quality-adjusted survival analysis. This consisted of defining four clinical states (toxicity, treatment free of toxicity, no treatment nor symptoms, relapse), then summing up the average times spent in each state weighted by utility coefficients that reflect relative value according to quality of life. The resulting quality-adjusted time without symptoms or toxicity (Q-TWIST) was compared between randomized groups, and sensitivity (threshold) analyses to the choice of utility coefficients was performed. Over 73 months after randomization, the fludarabine group gained a mean of 45 days of toxicity free survival at CAP, and 61 days over ChOP. The mean TWIST was 27.05 months with CAP, 31.5 months with ChOP and 32.95 months with fludarabine. The threshold analyses showed that, whatever the utility weights, the mean Q-TWIST was always greater with ChOP or fludarabine as compared to CAP. Fludarabine was consistently a better treatment than ChOP, except in the unlikely case of high utility weights attributed to toxicity and low utility weights attributed to treatment. Nevertheless, from a clinical point of view, differences between ChOP and fludarabine were moderate or event slight (mean difference in TWIST of 1.45 months). We conclude that patients with advanced CLL have a moderate benefit in terms of Q-TWIST when treated with fludarabine over ChOP. These two treatments are always superior to CAP. PMID- 11438418 TI - Histochemical characterization of primary capillary endothelial cells from porcine brains using monoclonal antibodies and fluorescein isothiocyanate labelled lectins: implications for drug delivery. AB - Primary endothelial cells isolated from cerebral microvessels by combined mechanical and enzymatic treatment from porcine brains were characterized with regard to identity, purity and membrane surface characteristics. Cells were grown in culture to adherent monolayers and characterized morphologically and histochemically by their binding for fluorescently-labelled lectins and monoclonal antibodies detected by indirect immunofluorescence. The binding patterns of the cells were compared with the affinity of frozen tissue sections of porcine brain cortex for the markers. Endothelial cells in culture were characterized by the binding of von Willebrand factor, vimentin and fibronectin antibodies. They failed to react with anti-glial fibrillary acid protein, anti galactocerebroside C and anti-neurofilament 160 antibodies characteristic for astrocytes, oligodendrocytes and neurons, respectively. Cell cultures were stained by the lectins, wheat germ agglutinin, horse gram agglutinin and soybean agglutinin, demonstrating the presence of N-acetylglucosamine and N acetylgalactosamine residues on membrane surface. Binding sites for concanavalin A and peanut agglutinin characteristic for mannose and galactose could not be detected. Cell age and differentiation had no effect on lectin and antibody staining. Cell cultures gave staining results similar to those of microvessels in frozen tissue sections. The results of morphology, antibody and lectin staining pattern indicate that our in vitro endothelial cell culture model retained many histological characteristics observed for capillary microvessels in vivo and appears to be suitable for studying uptake and targeting properties of drug carrier systems with regard to the blood-brain barrier. PMID- 11438419 TI - Stealth monensin immunoliposomes as potentiator of immunotoxins in vitro. AB - Stealth monensin liposomes (SML) were prepared using dipalmitoyl phosphatidylcholine, cholesterol, distearoyl glycerophosphoethanolamine coupled to polyethylene glycol, stearylamine, and N-succinimidyl pyridodithiopropionate linked to stearyl amine, in the molar ratio of 10:5:1.4:1.4:1.5. SML was conjugated to the anti-MY9 antibody by a disulfide linkage to form stealth monensin immunoliposomes (SMIL) by an already established procedure. The encapsulation concentrations of monensin in SML and SMIL were 10(-7) and 4.9x10( 8) M, respectively. More than 20% of monensin remained in circulation after 24 h in BALB/c mice. The ability of SML and SMIL to potentiate the effect of anti-MY9 immunotoxin (anti-MY9-IT) was tested against human leukemia HL-60 sensitive and resistant tumor cells in vitro. SML and SMIL potentiated the activity of anti-MY9 IT by 10-20 times against HL-60 sensitive tumor cell lines. However, greater potentiation of anti-MY9-IT was observed in combination with SML and SMIL against HL-60 resistant tumor cells, found to be 200 and 500 times, respectively. The potentiation of anti-MY9-IT by SMIL was more than two-fold compared with SML against both HL-60 sensitive and resistant tumor cells. Transmission electron microscopy studies conducted with HL-60 resistant cells incubated with anti-MY9 IT and monensin liposomes showed significant dilation of the golgi, which was reversible after re-incubation in fresh medium. Our studies show that SML and SMIL can be successfully used to potentiate the activity of ricin based anti-MY9 IT in vitro, and further in vivo studies will demonstrate the usefulness of this approach. PMID- 11438420 TI - Improved nasal absorption of drugs using poly-L-arginine: effects of concentration and molecular weight of poly-L-arginine on the nasal absorption of fluorescein isothiocyanate-dextran in rats. AB - The effects of the concentration and molecular weight of poly-L-arginine (poly-L Arg) on the in vivo nasal absorption of fluorescein isothiocyanate-labeled dextran (MW, 4 kDa, FD-4) in rats were studied. When poly-L-Arg with a range of different molecular weights (MW, 8.9, 45.5 and 92.0 kDa) was applied intranasally at various concentrations, the bioavailability (F(0-9 h)) of FD-4 increased with the increasing concentration of poly-L-Arg. The enhanced absorption was also dependent on the molar concentration, in that the poly-L-Arg with a higher molecular weight increased F(0-9 h) at a lower molar concentration. In addition, for each applied concentration, the poly-L-Arg exhibited a molecular weight dependence as far as the enhancement of FD-4 absorption was concerned. On the other hand, the maximum absorption rate (MAR) of FD-4, calculated by means of a deconvolution method, tended to reach a maximum plateau level at a lower applied concentration for the poly-L-Arg with the highest molecular weight, but this plateau level was almost the same for poly-L-Arg with molecular weights of 45.5 and 92.0 kDa. Moreover, the simulated absorption profiles of FD-4 indicate that the degree of enhancement (the level of MAR and the subsequent reduction in the absorption rate) was dependent on the molecular weight of poly-L-Arg, while the effect of poly-L-Arg was maintained for a longer period, depending on the applied concentration, although the MAR was relatively similar. These results indicate that the molecular weight of poly-L-Arg appears to affect both the enhancing efficiency (absorption rate) and the time-frame of this enhancing effect, whereas the concentrations of each poly-L-Arg system applied only have an effect on the time-frame. These effects may also be associated with the charge density of a poly-L-Arg molecule. PMID- 11438421 TI - The influence of active secretion processes on intestinal absorption of salbutamol in the rat. AB - Salbutamol was perfused in the small intestine of rat using a standard rat gut "in situ" preparation: (1) in inhibitor-free solution at seven different concentrations (0.15, 0.29, 1.20, 5.0, 9.0, 13.0 and 18.0mM); (2) at a 0.29mM concentration - thought to be close to the allometric dose in man - in the presence of a non-specific enzyme inhibitor, sodium azide (0.3, 3.0 and 6.0mM); and (3) at 0.29mM in the presence of a selective secretion inhibitor, verapamil (10.0 and 20.0mM). In free solution, the mixed-order rate constants, k'(a), of salbutamol increase as the solute concentration increases until an apparent asymptotic value is reached. This could be due to the saturation of enzymatic systems responsible for the secretion of the drug from the enterocyte to the luminal fluid, a process that could explain the poor absorption of salbutamol. In the presence of sodium azide, the k(a) values increased about 1.5-fold, whereas in the presence of verapamil they increased two- to three-fold. These results indicate that salbutamol can act as a substrate of an intestinal secretory transport, which probably includes--at least in part--the enzyme P-glycoprotein, since verapamil has been shown to inhibit this enzyme by dose-dependent competition. This leads to a secretion-limited peroral absorption of salbutamol, which contributes to the poor oral bioavailability of the drug. The possible options for improving salbutamol absorption are discussed. PMID- 11438422 TI - Microparticles for delivering therapeutic peptides and proteins to the lumen of the small intestine. AB - Several different peptides and proteins, such as the pancreatic trypsin inhibitor, growth factors and trefoil peptides, are known to play important roles in maintaining the structure and function of the gastrointestinal wall. With the advent of recombinant biotechnology, it has become feasible to test some of these proteins as therapeutics in different inflammatory conditions of the intestines. However, the harsh pH and enzymatic conditions of the stomach can lead to their inactivation. This research was aimed at the development of particulate, gastric resistant pharmaceutical forms, incorporating those bioactive molecules. Mixtures of proteins in powder form were coated with cellulose acetate phthalate, Eudragit S100 or Eudragit RS PO, using simple preparation techniques based on single emulsion/solvent evaporation. Using aprotinin as a model drug, it was found that these procedures were effective in microencapsulating protein in the solid form without affecting its biological activity. Furthermore, and in particular with the first two polymers above, particles showed adequate in vitro release patterns for the applications envisioned. PMID- 11438423 TI - In-vitro comparative study of buccal mucoadhesive performance of different polymeric films. AB - A comparison of the buccal mucoadhesive performance of different polymeric films was carried out using texture analyzer TA-XT2i. A large range of putative polymers differing in their chemical nature, molecular structure as well as hydration status was used. The used polymeric films were classified in rank order of buccal mucoadhesive performance, namely carbopol 971P>polycarbophil>Carrrageenan type lambda > Sodium carboxymethylcellulose. Swelling state as well as tensile strength of the used polymeric films was used as measuring parameters of mucoadhesive interaction. These two approaches gave two opposite orders of performance between CMC and Carrrageenan type lambda after a contact time of 15 min. However the measurement of the viscoelastic moduli of the hydrogels gave the same ranking order of mucoadhesive performance after the same contact time. In reference to the previous works, we noted the importance of the molecular weight, the density of charges, the composition of which the chains of molecules are capable to arrange themselves in a network like form, thus those which are characterized by a tan delta<1 (i.e network formation), are those which develop the best synergism with the mucus because of the reinforcement of an established link. The goal of this study is to assess the buccal mucoadhesive performance aiming to optimize the design of drug delivery via buccal mucoadhesive polymeric films PMID- 11438424 TI - Gender- or age-related binding characteristics of valproic acid to serum proteins in adult patients with epilepsy. AB - The aim of the present study was to determine the gender- or age-related binding characteristics of valproic acid (VPA) to serum proteins in the adult population. Serum samples examined in the study were obtained from 70 adult patients (36 males, 34 females) with epilepsy on VPA monotherapy. Their age ranged from 16 to 68 years (mean age with (SD), 37.7 (15.7) years; <45 years, n=44; >/=45 years, n=26). The in vivo population binding parameters of VPA to serum proteins and theoretical minimal unbound serum VPA fraction (Fu) were determined using an equation derived from the Scatchard equation in: (1), all; (2), male and female subgroups; and (3), younger (<45 years) and older (>/=45 years) subgroups. There was a significant difference in serum concentration of unbound VPA between male and female patients. The mean association constant (K) was 0.010 microM(-1) in all, male, and female patients. The mean total concentration of binding sites (n(Pt)) was 1453 microM for all patients, and 1561 and 1394 microM for male and female patients, respectively. The Fu was 0.064 for all patients, and 0.060 and 0.067 for male and female patients, respectively. There were no significant differences in the binding characteristics of VPA to serum proteins between the male and female groups. On the other hand, there were significant differences in the serum albumin concentration and molar concentration ratio of free fatty acids to albumin in serum between the younger and older patients. The mean value of K was 0.016 microM(-1) for the younger patients and 0.007 microM (-1) for the older patients. The mean n(Pt) was 1157 microM for the younger patients and 1703 microM for the older patients. The Fu was 0.051 for the younger patients and 0.077 for the older patients. Thus, significant differences were observed in the binding characteristics of VPA to serum proteins between the younger and older groups. Our results show that age, but not gender, has significant influences on the binding characteristics of VPA to serum proteins in our patient population. PMID- 11438425 TI - Complexation of phenytoin with some hydrophilic cyclodextrins: effect on aqueous solubility, dissolution rate, and anticonvulsant activity in mice. AB - The main objective of this study was to evaluate the influence of hydroxypropylated beta- and gamma-cyclodextrins and Me-beta-cyclodextrin (HP-beta CD, HP-gamma-CD, and Me-beta-CD, respectively) on the dissolution rate and bioavailability of the antiepileptic agent, phenytoin (DPH). The corresponding solid complexes were prepared by a freeze-drying method and characterized by infrared spectroscopy, X-ray powder diffraction, and differential scanning calorimetry studies. Evidence of inclusion complex formation in the case of HP beta-CD was obtained by (1)H- and (13)C-nuclear magnetic resonance spectroscopy. Drug solubility and dissolution rate in 0.05 M potassium phosphate buffer (pH 6) were notably improved by employing the beta-CDs. Thus a 45% w/v HP-beta-CD or Me beta-CD solution gave rise to an increase of dissolved drug of 420- and 578-fold, respectively. The Q(10) (i.e. percentage of dissolved DPH at 10 min) was 5.2% for the pure drug and 93, 98, and 96% for DPH/HP-beta-CD, DPH/HP-gamma-CD, and DPH/Me beta-CD complexes, respectively. Moreover, it was found that in the maximal electroshock seizure test in mice the DPH/Me-beta-CD complex exhibited anticonvulsant activity similar to DPH sodium salt (NaDPH). PMID- 11438426 TI - Effect of anionic polymers on the release of propranolol hydrochloride from matrix tablets. AB - Anionic polymers, namely Eudragit S, Eudragit L 100-55, and sodium carboxymethylcellulose, were incorporated into hydroxypropylmethylcellulose (HPMC K100M) to modify the drug release from HPMC matrices. The effects of changing the ratio of HPMC to anionic polymers were examined in water and in media with different pH. The dissolution profiles were compared according to release rates. The interaction between propranolol hydrochloride and anionic polymers was confirmed using the UV difference spectra method. The drug release was controlled with the type of anionic polymer and the interaction between propranolol hydrochloride and anionic polymers. The HPMC-anionic polymer ratio also influenced the drug release. The matrix containing HPMC-Eudragit L 100-55 (1:1 ratio) produced pH-independent extended-release tablets in water, 0.1 N HCl, and pH 6.8 phosphate buffer. PMID- 11438427 TI - Trehalose-hydroxyethylcellulose microspheres containing vancomycin for topical drug delivery. AB - A new formulation, in which vancomycin is entrapped into trehalose and hydroxyethylcellulose (Natrosol) spherical matrices, is described. Microspheres were produced by the solvent evaporation method. The entrapped drug was fully recovered following microspheres dissolution. Differential scanning calorimetry analyses proved that Natrosol maintains trehalose in its amorphous form. The stabilizing effects of trehalose on vancomycin were evaluated even after long storage and heating of microspheres. Calorimetric data indicated no decomposition of the entrapped drug. In vitro drug release, already performed by using a general two-compartment linear time-invariant open model, suggests that the new delivery system is suitable for topical application on extensive and purulent or burn wounds, when the skin is heavily damaged and the barrier disrupted. The system activation is determined by osmotic phenomena. The prepared new delivery system seems to have characteristics suitable for topical applications on extensive and purulent wounds. The system is able to take away serous exudates from wounds, thus letting the matrix to swell and form a viscous gel-like dispersion that, in turn, enables drug diffusion. PMID- 11438428 TI - Porous biodegradable microparticles for delivery of pentamidine. AB - The primary objective of this study was to develop a method for the preparation of porous biodegradable controlled release formulation of poly(lactide/glycolide) (PLGA). The model drug used for this study was pentamidine. Scanning electron microscopy pictures showed that these microparticles are highly porous and spherical in shape. A comparison of particle size reveals a similar median particle size (54-68 microm) in all six batches. The particles are all smaller than 90 microm. Differential scanning calorimetry thermograms revealed that pentamidine was mostly present in the crystalline form in the microparticles and did not dissolve in PLGA. The efficiency of encapsulation of pentamidine was higher than 58% in all six batches. The amount of drug released from these microparticles was at least 12% within the first 60 min. At least 50% of the total drug was released within the first 4 h. Drug release from these microparticles continued for up to 12 h. This faster drug dissolution was due to the highly porous surface. This highly porous surface will allow large molecules to release at a much faster rate than the regular microcapsules/microspheres. PMID- 11438429 TI - Purification and identification of major soluble 40-kDa antigenic protein from Entamoeba histolytica: its application for serodiagnosis of asymptomatic amebiasis. AB - One of the major soluble antigenic proteins of Entamoeba histolytica was purified to homogeneity and identified on a molecular basis. Its recombinant protein was expressed in Escherichia coli as a fusion protein with Shistosoma japonicum glutathione S-transferase. Apparent molecular weight of the purified antigenic protein was estimated to be 40-kDa and molecular-based analysis indicated that the purified protein was NADP+-dependent alcohol dehydrogenase (EhADH1). The application of the purified protein for the serodiagnosis of amebiasis was evaluated using an enzyme-linked immunosorbent assay applied to sera obtained from patients with amebiasis and healthy human controls. The purified protein was well recognized by the sera from asymptomatic amebiasis humans (22/22, 100%), whereas, it was less recognized by the sera from symptomatic amebiasis patients (5/16, 31%) with amebic colitis or liver abscess. To confirm the antigenicity of EhADH1, the recombinant glutathione S-transferase-EhADH1 fusion protein was also evaluated by the enzyme-linked immunosorbent assay using the same sera. The recombinant protein was also recognized by the sera from asymptomatic amebiasis humans (14/22, 64%) and less recognized by the sera from symptomatic amebiasis patients (2/16, 13%). These results suggest that the purified protein is applicable antigen for serodiagnostic screening of asymptomatic amebiasis humans. PMID- 11438430 TI - A single intradermal administration of soluble leishmanial antigen and plasmid expressing interleukin-12 protects BALB/c mice from Leishmania major infection. AB - In murine leishmaniasis, the induction of the T-helper type 1 (Th1) response contributes to infection resistance, whereas the establishment of the Th2 response makes the mice susceptible to infection. Interleukin-12 (IL-12) plays a pivotal role in the diversification of immune responses to the Th1 type. In this study, we tested whether the co-administration of IL-12 expression plasmid which compose p35 and p40 subunits and soluble leishmanial antigen (SLA) will skew the susceptible BALB/c mice to Th1 response and protect from leishmaniasis. When the mice were intradermally injected with the combination of IL-12 plasmid and SLA 7 days prior to the challenge with 1x10(6) promastigotes of Leishmania major, the local lesions completely healed and the parasite burden in the local lymph nodes significantly decreased. The cured mice attained long-term immunity, and were resistant to any subsequent rechallenge of the lethal dose of the parasite. The protective effect was associated with the development of a Th1 response, as demonstrated by the enhanced level of antigen-specific interferon-gamma (IFN gamma) and dominant production of IgG2a in the serum. In contrast, the administration of empty plasmid plus SLA or IL-12 plasmid alone failed to protect the disease and shape the Th1 response. Furthermore, the protective efficiency induced by the vaccination was clearly prevented by the injection of either neutralizing anti-IL-12 mAb or anti-IFN-gamma mAb. The IL-12 expression plasmid is thus an effective adjuvant for the elicitation of a protective Th1 response against leishmaniasis and is therefore, considered to be appropriate for vaccinations that require the induction of Th1 type immunity. PMID- 11438431 TI - Phylogenetic identification of Sparganum proliferum as a pseudophyllidean cestode by the sequence analyses on mitochondrial COI and nuclear sdhB genes. AB - Sparganum proliferum is a larval cestode for which the adult stage is unknown. It is characterized by the continuous branching and budding when parasitized to humans, and causes fatal human sparganosis. However, the biological features of S. proliferum, including its taxonomic status, still remain obscure. Our previous investigation suggested that S. proliferum might be phylogenetically distinct from Spirometra erinaceieuropaei, by the analysis on mitochondrial NADH dehydrogenase subunit 3 (ND3) gene. However, mitochondrial DNA sequence in Platyhelminth is known to have heteroplasmy within a species. Therefore, in the present study, we have investigated the complete nucleotide sequences of mitochondrial cytochrome c oxidase subunit I (COI) gene and the partial nucleotide sequences of nuclear coded succinate dehydrogenase iron-sulfur protein subunit gene (sdhB). The results clearly demonstrated that S. proliferum is a distinct species from S. erinaceieuropaei, and that S. proliferum belongs to the order Pseudophyllidea. PMID- 11438432 TI - Molecular cloning and characterization of cathepsin L encoding genes from Fasciola gigantica. AB - In this study cDNAs encoding cathepsin L-like proteins of Fasciola gigantica were cloned by the reverse transcription polymerase chain reaction method (RT-PCR) from total RNA of adult specimens. DNA sequence analyses revealed that six different cathepsin L cDNA fragments were isolated, which have DNA sequence identities of 87-99% towards the homologous genes from F. hepatica. Gene expression was studied at the RNA level by Northern and RNA in situ hybridizations. Northern analysis showed the cathepsin L genes to be strongly expressed in adult parasites as a group of 1050 nt sized RNAs. RNA in situ hybridization localized cathepsin L RNA to the cecal epithelial cells. Southern hybridization was used to determine the number of cathepsin L genes and indicated the presence of a family of closely related cathepsin L genes in the genome of F. gigantica. PMID- 11438433 TI - Element concentration variability in the whaleworm Anisakis simplex s.l. AB - Concentrations of the elements C, N, Fe, Na, K, Ca and Mg were analysed by chromatography and atomic absorption spectrometry in the whaleworm Anisakis simplex sensu lato. Nearly all mean inorganic element concentrations measured were higher in adult worms than in larval forms. Similarly, adults contained significantly higher nitrogen (i.e. lower C/N ratio values) than either eggs or larvae. Evidence for the significance of the parasitic life cycle stage and the systematic position of their hosts on the element concentration variability is discussed. PMID- 11438434 TI - First report of Trichinella nativa in red foxes (Vulpes vulpes schrencki) from Otaru City, Hokkaido, Japan. AB - Forty-three red foxes (Vulpes vulpes schrencki) and nine raccoon dogs (Nyctereutes procyonoides viverrinus) were captured in Otaru City, Hokkaido, Japan and examined by muscle digestion for the presence of Trichinella sp. larvae. Of the foxes, five (11.6%) were positive for larvae of Trichinella nativa while none of the raccoon dogs were found to be positive. This finding suggests that the red foxes are important reservoir hosts of sylvatic trichinellosis in Otaru, Hokkaido. This is the first report on Trichinella nativa in wild red foxes of Hokkaido, Japan. PMID- 11438435 TI - Immunolocalization of the 38.3 kDa calponin-like protein in stratified muscles of the tail of Schistosoma japonicum cercariae. AB - Calponins are proteins present in vertebrate smooth musculature where they occur in association with thin myofilaments. Calponins are not present in vertebrate or invertebrate striated muscles. The blood fluke Schistosoma japonicum expresses a 38.3-kDa protein that bears substantial homology with vertebrate calponin and occurs entirely within smooth musculature of adults. Calponin-like immunoreactivity has been demonstrated in smooth muscles of many invertebrate phyla. The Schistosoma japonicum calponin has been localised in smooth myofibrils of adults where it is associated with myofilaments and sarcoplasmic reticulum. In this study, the ultrastructural localisation of the protein in muscles of S. japonicum cercariae is described. The protein is present in smooth muscles of the forebody and the stratified muscle of the tail. Within the stratified layer, the protein occurs predominantly in transverse arrays of sarcoplasmic reticulum. The localisation data suggest that the calponin-like protein of S. japonicum is involved in contraction of the stratified tail muscle. Furthermore, the presence of a calponin system in the stratified muscle suggests that this muscle is simply a superior form of muscle, closely related to smooth muscles that use a caldesmin calponin system in contraction. PMID- 11438436 TI - Detection of kinetoplast DNA of Trypanosoma cruzi from dried feces of triatomine bugs by PCR. AB - It is important to clarify the distribution of infected triatomine bugs in the endemic area of Chagas' disease for proper control. In the present study, we tried to detect T. cruzi kinetoplast DNA by PCR from dried triatomine feces collected from the house wall of an endemic area to assess the distribution of infected bugs more easily. The primers (P35/P36) were chosen to amplify the conserved region within the minirepeats of T. cruzi kinetoplast minicircle DNA. The kinetoplast DNA of T. cruzi could be actually detected in the dried feces collected from the wall of a brick-built house in Santa Cruz, Bolivia. Next, we examined the stability of T. cruzi kinetoplast DNA in the feces exposed to artificial environments. T. cruzi DNA was also detected by PCR in the feces left for 26 weeks at 25 degrees C and in those left for 4 weeks at 40 degrees C. The present study indicates that examination of dried feces on the wall can be an effective tool for surveillance of the natural infection of triatomine bugs that live in houses. PMID- 11438437 TI - Cytokine and chemokine mRNA expression in neutrophils from CBA/NSlc mice infected with Plasmodium berghei ANKA that induces experimental cerebral malaria. AB - To investigate the role of neutrophils in experimental cerebral malaria (ECM), in a previous study we found that early neutrophil depletion prevented the development of ECM and down regulated the expression of Th1 cytokines in the brain. To further clarify the mechanisms responsible for these findings, in the present study, using RT-PCR, we examined the expression of cytokine and chemokine mRNAs in neutrophils and macrophages after PbA infection. We found that, after infection, neutrophils not only expressed cytokines IL-2, IL-12p40, IL-18, IFN gamma and TNF-alpha mRNAs, but also mRNAs for Th1 chemoattractive chemokines, monokine-induced by IFN-gamma (MIG), macrophage-inflammatory protein-1alpha (MIP 1alpha) and IFN-gamma inducible protein-10 (IP-10). Neutrophil depletion down regulated the expression of IL-18 and MIG mRNAs in macrophages, but did not affect the expression of IFN-gamma, TNF-alpha, MIP-1alpha and IP-10 mRNAs. Therefore, this study confirms our hypothesis that neutrophils may play a role in the pathogenesis of ECM via their expression of cytokines or chemokines. PMID- 11438438 TI - Intraspecific variability among NADH dehydrogenase subunit 1 sequences of Taenia hydatigena. AB - This paper describes intraspecific variability of the partial sequences of the mitochondrial ND1 gene among isolates of Taenia hydatigena from pigs in Poland, Ukraine and Wales. The differences between studied isolates ranged from 0.4 to 5.5%, which exceeds the variability within the same fragment between the different genetic variants of Echinococcus multilocularis and is comparable with the variability between the most closely related strains (G5/G6/G7) of E. granulosus. The biggest difference (5.5%) was found between the geographically most distant Ukrainian and Welsh samples of T. hydatigena while the samples collected from the neighbouring locations in Poland, were most similar to each other. PMID- 11438439 TI - A histochemical study of the secretory gland cells of Cercaria shikokuensis and their role during development from cercaria to metacercaria. AB - The roles of secretory glands during the developmental process from an immature cercaria to a metacercaria in Cercaria shikokuensis were studied. Four types of secretory cells were identified in this species. On maturation of the cercaria in redia, the products of ventral gland cells and mucoid gland cells formed a thick surface coat on the mature cercaria, and the products of cephalic gland cells also formed a thin cover on the surface coat. In the process leading to the formation of a metacercaria, the surface coat constituted the outer layer of the cyst, mucoid gland cells secreted mucous substances inside the wall, and then cystogenous gland cells discharged their products to the inner wall. The cyst wall was composed of four layers, and it was thought that the outermost surface layer helped the cyst wall to adhere to the matrix and the intermediate layers helped to put together outer and inner walls. PMID- 11438440 TI - Calcium and the control of mammalian cortical granule exocytosis. AB - At fertilization, the release of intracellular calcium is necessary and sufficient for most, if not virtually all, of the major events of egg activation that are responsible for the onset of embryonic development. In mammalian eggs, repetitive calcium oscillations stimulate egg activation events through calcium dependent effectors, such as calmodulin, protein kinases, and specific proteins involved in exocytosis. One of the earliest calcium-dependent events is the exocytosis of cortical granules (CGs), a secretory event resulting in the block to polyspermy and the prevention of triploidy. Emerging studies suggest that CG release in mature eggs is dependent upon calcium-dependent proteins similar to those in somatic cells employed to undergo calcium-regulated exocytosis. In contrast, pre-ovulatory oocytes are incompetent to undergo CG exocytosis due to deficiencies in the ability to release and respond to increases in intracellular calcium. The development of competence to release and respond to calcium is relevant to both animal and human in vitro fertilization programs that largely utilize ovarian oocytes not all of which are fully activation competent. PMID- 11438441 TI - Gene targeting in hemostasis: protein C. AB - Protein C (PC) is the zymogen form of a serine protease, activated protein C (APC), a naturally occurring anticoagulant. In control of the coagulation of blood, APC functions by attenuating thrombin formation. It serves this role through inactivation, by limited proteolysis, of two important cofactors for overall clot formation, one of which, Factor Va (FVa), stimulates prothrombin activation, and another, Factor VIIIa (FVIIIa), enhances activation of coagulation Factor X (FX). In maintaining the fluidity of blood, APC also indirectly functions in fibrinolysis, in one manner by directly inactivating an inhibitor of plasminogen activation, plasminogen activation inhibitor-1 (PAI-1), and in another manner via its role in attenuating thrombin production, with the resulting effect of limiting production of another thrombin-dependent fibrinolytic inhibitor, thrombin activatable fibrinolysis inhibitor (TAFI). PC, and other components of the PC anticoagulant pathway, e.g., protein S (PS), thrombomodulin (Tm), and endothelial cell protein C receptor (EPCR), also can serve as anti-inflammatory mediators, through a number of different thrombin dependent and thrombin-independent mechanisms. A large number of symptomatic and asymptomatic mutations occur in PC in humans, which express a variety of phenotypes. Generation and characterization of a murine model of a total PC gene inactivation has demonstrated that while an untreated total PC deficiency results in neonatal death through DIC-related abnormalities, a valuable resource is now available to study phenotypes of less severe deficiencies of this protein. Such studies will lead to advances in an understanding of the relative role of this protein system in the various pathways in which it has an influence. PMID- 11438442 TI - PKC in eggs and embryos. AB - After a spermatozoon enters the egg, rapid processes such as the cortical granules exocytosis (CGE) occur. Other, later processes are observed as well, and they include resumption of the second meiotic division, second polar body (PBII) extrusion and rearrangement of the DNA inside the pronuclei (PN). The above mentioned biochemical processes involve signal transduction pathways that are well known in other cell systems, and require mediation of second messengers like Ca2+ and diacylglycerol (DAG) which are protein kinase C (PKC) activators. The present review, based upon recently published studies, raises the possibility of PKC involvement in fertilization and in early developmental stages of the mammalian embryo. PMID- 11438443 TI - Vitamin D and cancer: effects of 1,25(OH)2D3 and its analogs on growth control and tumorigenesis. AB - Today, it is well established that besides playing a crucial role in the establishment and maintenance of the calcium homeostasis in the body, the active form of vitamin D, 1,25(OH)2D3, also acts an effective regulator of cell growth and differentiation in a number of different cell types, including cancer cells. This has led to an increased interest in using 1,25(OH)2D3 in the treatment or prevention of cancer patients and to a substantial number of studies investigating the effect of 1,25(OH)2D3 on cancer cells. The results are encouraging, but clearly demonstrate that the therapeutic window of 1,25(OH)2D3 is extremely narrow due to the calcemic adverse effects of this compound. Much effort has consequently been directed into identifying vitamin D analogs with potent cell regulatory effects but with weaker effects on the calcium metabolism than those of 1,25(OH)2D3. In an attempt to clarify the mechanisms implicated in the cell regulatory effects of 1,25(OH)2D3 and eventually facilitate the process of developing new specific vitamin D analogs, numerous investigations have been carried out with 1,25(OH)2D3 and its analogs. The present review will focus on the results obtained in these studies and describe some of the synthetic analogs, which have shown to be of particular interest in relation to cancer. PMID- 11438445 TI - Kenneth Merle Brinkhous, M.D. 1908-2000. PMID- 11438446 TI - Cardiac myosin and the TH1/TH2 paradigm in autoimmune myocarditis. PMID- 11438447 TI - Estrogen receptor beta in prostate cancer: brake pedal or accelerator? PMID- 11438448 TI - Circulating breast cancer cells are frequently apoptotic. AB - Automatic search for cytokeratin/mucin-1 double immunofluorescence was performed to detect and characterize circulating epithelial tumor cells in patients with advanced breast cancer. The peripheral blood samples in 8 of 19 patients (42.1%) presented with cytokeratin-positive and epithelial-type mucin-positive (CK(+)/MUC1(+)) tumor cells. Detailed microscopic analysis, however, suggested that the majority of the double immunopositive cells was apoptotic according to an "inclusion type" cytokeratin staining pattern and nuclear condensation. Furthermore, apoptosis-related DNA strand breaks could be demonstrated by applying the TdT-uridine nick end labeling assay in these cells. In 3 of 8 positive samples all of the CK(+)/MUC1(+) cells displayed apoptotic features. We conclude that apoptotic cells significantly contribute to the circulating tumor cell fraction in breast cancer patients. As the predictive value of such cells for the outcome of the disease is unclear, they should be considered separately when analyzing tumor cell dissemination. PMID- 11438449 TI - Metallothionein isoform 3 overexpression is associated with breast cancers having a poor prognosis. AB - The third isoform (MT-3) of the metallothionein gene family is unique in that it has a limited tissue distribution, is not induced by metals, has a neuronal growth inhibitory activity, and sequesters zinc more effectively under zinc depleted conditions. The goal of the present study was to determine whether MT-3 was absent in normal breast tissue, was overexpressed in breast cancers, and if MT-3 overexpression would be associated with disease outcome. A combination of immunohistochemistry and reverse-transcription polymerase chain reaction was used to demonstrate that the normal breast had no detectable expression of MT-3 mRNA or protein. Using immunohistochemistry, it was shown that MT-3 was overexpressed in 25 of 34 cases of breast cancer. In all cases of positive staining, MT-3 was diffusely localized to the cytoplasm. The tumors from these 34 cases were divided as to outcome based on known 5-year survival, with 20 patients being disease free at 5 years (good outcome) and the other 14 having recurring disease within 5 years (bad outcome). When analyzed for MT-3 staining, it was shown that there was a trend for increased MT-3 immunoreactivity in the group having bad outcomes. However, when the tumor subgrouping was further defined on the basis of carcinoma in situ (CIS), there was a marked significant difference in MT-3 staining between patients with good and bad outcomes. Limited to DCIS, MT-3 staining was significantly increased in patients with bad outcomes compared to those with good outcomes. Thus, these studies demonstrate that MT-3 is overexpressed in selected breast cancers and that overexpression is associated with tumors having a poor prognosis. PMID- 11438450 TI - Induction of c-Met proto-oncogene by Epstein-Barr virus latent membrane protein-1 and the correlation with cervical lymph node metastasis of nasopharyngeal carcinoma. AB - Nasopharyngeal carcinoma (NPC) is distinctive in head and neck carcinomas for its close association with Epstein-Barr virus and its highly metastatic nature. Up regulation of cell motility is essential for enhancement of metastatic potential. The expression of c-Met proto-oncogene, a high-affinity receptor for hepatocyte growth factor/scatter factor, has been reported to correlate with metastatic ability of the tumor cell. We observed close association of c-Met expression with cervical lymph node metastasis (P = 0.0272) in 39 NPC specimens studied immunohistochemically. Epstein-Barr virus-encoding latent membrane protein-1 (LMP 1) is a primary oncogene and is suggested to enhance the metastatic property of NPC. Previously, we reported that LMP-1 enhanced the motility of Madin-Darby canine kidney (MDCK) epithelial cells that was mediated by activation of Ets-1 transcription factor. Therefore, we examined the interrelationships of LMP-1, Ets 1, and c-Met. In immunohistochemical studies, the expression of LMP-1, Ets-1, and c-Met correlated significantly with each other in NPC (LMP-1 versus Ets-1, P < 0.0001; Ets-1 versus c-Met, P = 0.0012; LMP-1 versus Met, P = 0.0005). Transfection of LMP-1-expressing plasmid in MDCK cells induced c-Met protein expression. The c-Met protein was also induced by Ets-1 expression, and induction of c-Met by LMP-1 was suppressed by introducing a dominant-negative form of Ets-1 in LMP-1-expressing MDCK cells. These results suggest that LMP-1 induces c-Met through the activation of Ets-1, which may contribute in part to the highly metastatic potential of NPC. PMID- 11438451 TI - Allelic analysis of serous ovarian carcinoma reveals two putative tumor suppressor loci at 18q22-q23 distal to SMAD4, SMAD2, and DCC. AB - The distal half of chromosome arm 18q is frequently lost in ovarian carcinoma. To define the putative tumor suppressor locus/loci more precisely we performed allelic analysis with 27 polymorphic microsatellite markers located at 18q12.3 q23 in 64 serous and 9 mucinous ovarian carcinomas. Fifty-nine percent of the serous carcinomas, but only one (11%) of mucinous carcinomas, showed allelic loss at one or more loci (P = 0.018). In serous carcinomas, deletions were found to be associated with tumor grade and poor survival. The highest frequency of losses was detected at the distal part, 18q22-q23. Two minimal common regions of loss (MCRL) were identified at this region: MCRL1 between D18S465 and D18S61 at 18q22 (3.9 cM) and MCRL2 between D18S462 and D18S70 at 18q23 (5.8 cM). At 18q21.1, proximal to the MCRLs, there are three candidate tumor suppressor genes: SMAD4 (DPC4), SMAD2, and DCC. Their protein expression was studied by immunohistochemistry in normal ovarian tissue and serous carcinomas. Lost or very weak expression of SMAD4, SMAD2 and DCC was found in 28, 28, and 30% of serous carcinomas, respectively. Comparison of allelic loss and protein expression status indicated that none of these genes alone could be the target for the frequent allelic loss at 18q21.1. Together, these genes may account for a substantial proportion of the events, but not all of them. Thus, we propose that the frequent allelic loss at 18q is because of the effect of multiple genes, and there is at least one as yet unidentified tumor suppressor gene at 18q residing distal to SMAD4, SMAD2, and DCC involved in serous ovarian carcinoma. PMID- 11438452 TI - Reduced expression of plakoglobin correlates with adverse outcome in patients with neuroblastoma. AB - Plakoglobin and its homologue beta-catenin are cytoplasmic proteins that mediate adhesive functions by interacting with cadherin receptors and signaling activities by interacting with transcription factors. It has been suggested that plakoglobin can suppress tumorigenicity whereas beta-catenin can act as an oncogene. We investigated the correlation between the expression pattern of N cadherin, beta-catenin, and plakoglobin and tumor behavior in primary tumors of 20 neuroblastoma patients of all stages and in 11 human neuroblastoma cell lines. N-cadherin and beta-catenin were detected in 9 of 11 and 11 of 11 cell lines, respectively, whereas plakoglobin was undetectable or severely reduced in 6 of 11 cell lines. Tumor cells from 16 of 20 patients expressed N-cadherin and 20 of 20 patients expressed beta-catenin at levels similar to those of normal ganglion cells. Plakoglobin was undetectable in 9 of 20 tumors. Plakoglobin deficiency in the primary tumors was significantly associated with adverse clinical outcome. Five of the patients with plakoglobin-negative tumors died whereas four patients are alive without evident disease. In contrast, all patients with plakoglobin positive tumors are alive; 2 of 11 are alive with the disease and 9 of 11 are alive without evident disease. These results suggest that down-regulation of plakoglobin may be of prognostic value for neuroblastoma patients as predictor of poor outcome. PMID- 11438453 TI - Glial implants in gliomatosis peritonei arise from normal tissue, not from the associated teratoma. AB - Metaplasia of subcoelomic mesenchyme has been implicated, but not proven, in the pathogenesis of common gynecological diseases such as endometriosis and rarer entities such as leiomyomatosis peritonealis disseminata and gliomatosis peritonei (GP). GP is associated with ovarian teratomas and is characterized by numerous peritoneal and omental implants composed of glial tissue. Two theories to explain the origin of GP have been proposed. In one, glial implants arise from the teratoma, whereas in the other, pluripotent Mullerian stem cells in the peritoneum or subjacent mesenchyme undergo glial metaplasia. To address the origin of GP, we exploited a unique characteristic of many ovarian teratomas: they often contain a duplicated set of maternal chromosomes and are thus homozygous at polymorphic microsatellite (MS) loci. In contrast, DNA from matched normal or metaplastic tissue (containing genetic material of both maternal and paternal origin) is expected to show heterozygosity at many of these same MS loci. DNA samples extracted from paraffin-embedded normal tissue, ovarian teratoma and three individual laser-dissected glial implants were studied in two cases of GP. In one case, all three implants and normal tissue showed heterozygosity at each of three MS loci on different chromosomes, whereas the teratoma showed homozygosity at the same MS loci. Similar results were observed in the second case. Our findings indicate that glial implants in GP often arise from cells within the peritoneum, presumably pluripotent Mullerian stem cells, and not from the associated ovarian teratoma. This finding has important implications for more common gynecological entities with debatable pathogenesis, such as endometriosis, by definitively demonstrating the metaplastic potential of stem cells within the peritoneal cavity. PMID- 11438454 TI - NF1 deletions in S-100 protein-positive and negative cells of sporadic and neurofibromatosis 1 (NF1)-associated plexiform neurofibromas and malignant peripheral nerve sheath tumors. AB - Although plexiform neurofibroma (PN) is thought to represent a benign neoplasm with the potential for malignant transformation (malignant peripheral nerve sheath tumor; MPNST), its neoplastic nature has been difficult to prove due to cellular heterogeneity, which hampers standard molecular genetic analysis. Its mixed composition typically includes Schwann cells, fibroblasts, perineurial-like cells, and mast cells. Although NF1 loss of heterozygosity has been reported in subsets of PNs, it remains uncertain which cell type(s) harbor these alterations. Using a dual-color fluorescence in situ hybridization and immunohistochemistry technique, we studied NF1 gene status in S-100 protein-positive and -negative cell subpopulations in archival paraffin-embedded specimens from seven PNs, two atypical PNs, one cellular/atypical PN, and eight MPNSTs derived from 13 patients, seven of which had neurofibromatosis type 1 (NF1). NF1 loss was detected in four of seven PNs and one atypical PN, with deletions entirely restricted to S-100 protein-immunoreactive Schwann cells. In contrast, all eight MPNSTs harbored NF1 deletions, regardless of S-100 protein expression or NF1 clinical status. Our results suggest that the Schwann cell is the primary neoplastic component in PNs and that S-100 protein-negative cells in MPNST represent dedifferentiated Schwann cells, which harbor NF1 deletions in both NF1 associated and sporadic tumors. PMID- 11438455 TI - Rolling circle amplification: a new approach to increase sensitivity for immunohistochemistry and flow cytometry. AB - Immunohistochemistry is a method that can provide complementary diagnostic and prognostic information to morphological observations and soluble assays. Sensitivity, specificity, or requirements for arduous sample preparation or signal amplification procedures often limit the application of this approach to routine clinical specimens. Rolling circle amplification (RCA) generates a localized signal via an isothermal amplification of an oligonucleotide circle. The application of this approach to immunohistochemistry could extend the utility of these methods to include a more complete set of immunological and molecular probes. RCA-mediated signal amplification was successfully applied to the sensitive and specific detection of a variety of cell surface antigens (CD3, CD20, and epithelial membrane antigen) and intracellular molecules (vimentin and prostate-specific antigen) within a variety of routinely fixed specimens, as well as samples prepared for flow cytometry. RCA technology, which has an intrinsically wide dynamic range, is a robust and simple procedure that can provide a universal platform for the localization of a wide variety of molecules as a function of either antigenicity or nucleic acid sequence. The use of RCA in this way could enhance the use of markers of current interest as well as permit the integration of emerging information from genomics and proteomics into cell- and tissue-based analyses. PMID- 11438456 TI - HLA-G and NK receptor are expressed in psoriatic skin: a possible pathway for regulating infiltrating T cells? AB - Recent data have suggested that in psoriasis, the T-infiltrating cells could be submitted to regulatory pathways, possibly through natural killer receptors. HLA G binds to different natural killer receptors and is able to inhibit T-cell functions. Because this molecule is induced by interferon-gamma, a major cytokine in psoriasis, we asked whether HLA-G and its receptor might be expressed in this disease. Specific RNAs for HLA-G1 and HLA-G5 were consistently found in lesional skin specimens, soluble HLA-G5 transcripts being found only in psoriasis. HLA-G protein was found in all psoriatic sections, but never in normal skin controls. Double labeling demonstrated that HLA-G-positive cells were CD68(+), CD11c(+) macrophages. The NKR ILT2 was also present in psoriatic skin, the T CD4(+) infiltrating cells expressing indeed ILT2. The demonstration of HLA-G and ILT2 expression in psoriatic skin suggests that this pathway may act as an inhibitory feed back aimed to down-regulate the deleterious effects of T-cell infiltrate in this disease. PMID- 11438457 TI - Comparative studies of the estrogen receptors beta and alpha and the androgen receptor in normal human prostate glands, dysplasia, and in primary and metastatic carcinoma. AB - An antibody, GC-17, thoroughly characterized for its specificity for estrogen receptor-beta (ER-beta), was used to immunolocalize the receptor in histologically normal prostate, prostatic intraepithelial neoplasia, primary carcinomas, and in metastases to lymph nodes and bone. Comparisons were made between ER-beta, estrogen receptor-alpha (ER-alpha), and androgen receptor (AR) immunostaining in these tissues. Concurrently, transcript expression of the three steroid hormone receptors was studied by reverse transcriptase-polymerase chain reaction analysis on laser capture-microdissected samples of normal prostatic acini, dysplasias, and carcinomas. In Western blot analyses, GC-17 selectively identified a 63-kd protein expressed in normal and malignant prostatic epithelial cells as well as in normal testicular and prostatic tissues. This protein likely represents a posttranslationally modified form of the long-form ER-beta, which has a predicted size of 59 kd based on polypeptide length. In normal prostate, ER beta immunostaining was predominately localized in the nuclei of basal cells and to a lesser extent stromal cells. ER-alpha staining was only present in stromal cell nuclei. AR immunostaining was variable in basal cells but strongly expressed in nuclei of secretory and stromal cells. Overall, prostatic carcinogenesis was characterized by a loss of ER-beta expression at the protein and transcript levels in high-grade dysplasias, its reappearance in grade 3 cancers, and its diminution/absence in grade 4/5 neoplasms. In contrast, AR was strongly expressed in all grades of dysplasia and carcinoma. Because ER-beta is thought to function as an inhibitor of prostatic growth, androgen action, presumably mediated by functional AR and unopposed by the beta receptor, may have provided a strong stimulus for aberrant cell growth. With the exception of a small subset of dysplasias in the central zone and a few carcinomas, ER-alpha-stained cells were not found in these lesions. The majority of bone and lymph node metastases contained cells that were immunostained for ER-beta. Expression of ER-beta in metastases may have been influenced by the local microenvironment in these tissues. In contrast, ER-alpha-stained cells were absent in bone metastases and rare in lymph nodes metastases. Irrespective of the site, AR-positive cells were found in all metastases. Based on our recent finding of anti-estrogen/ER-beta mediated growth inhibition of prostate cancer cells in vitro, the presence of ER beta in metastatic cells may have important implications for the treatment of late-stage disease. PMID- 11438458 TI - Plasma membrane-associated pY397FAK is a marker of cytotrophoblast invasion in vivo and in vitro. AB - During human pregnancy specialized placental cells of fetal origin, termed cytotrophoblasts, invade the uterus and its blood vessels. This tumor-like process anchors the conceptus to the mother and diverts the flow of uterine blood to the placenta. Previously, we showed that the expression of molecules with important functional roles, including a number of extracellular matrix integrin receptors, is precisely modulated during cytotrophoblast invasion in situ. Here we exploited this observation to study the role of the focal adhesion kinase (FAK), which transduces signals from the extracellular matrix and recruits additional signaling proteins to focal adhesions. Immunolocalization studies on tissue sections showed that FAK is expressed by cytotrophoblasts in all stages of differentiation. Because extracellular matrix-induced integrin clustering results in FAK (auto)phosphorylation on tyrosine 397 (Y397FAK), we also localized this form of the molecule. Immunolocalization experiments detected Y397FAK in a subset of cytotrophoblasts near the surface of the uterine wall. To assess the functional relevance of this observation, we used an adenovirus strategy to inhibit cytotrophoblast expression of FAK as the cells differentiated along the invasive pathway in vitro. Compared to control cells transduced with a wild-type virus, cytotrophoblasts that expressed antisense FAK exhibited a striking reduction in their ability to invade an extracellular matrix substrate. When cytotrophoblast differentiation was compromised (hypoxia in vitro, preeclampsia in vivo), Y397FAK levels associated with the plasma membrane were strikingly lower, although total FAK levels did not change. Together our results suggest that (auto)phosphorylation of Y397 on FAK is a critical component of the signaling pathway that mediates cytotrophoblast migration/invasion. PMID- 11438459 TI - Deregulation of renal transforming growth factor-beta1 after experimental short term ureteric obstruction in fetal sheep. AB - Renal malformations are the commonest cause of chronic renal failure in children and they are often associated with urinary tract abnormalities that impair fetal urine flow. Up-regulation of transforming growth factor-beta1 (TGF-beta1) occurs after experimental postnatal urinary tract obstruction and we recently reported increased levels of TGF-beta1 in human renal malformations (Yang SP et al, Am J Pathol 2000, 157:1633-1647). These findings led us to propose that obstruction induced stretch of developing renal epithelia causes up-regulation of TGF-beta1, which then perturbs renal development. In this study, therefore, we examined expression of components of the TGF-beta1 signaling axis in a previously characterized ovine model of fetal short-term urine flow impairment in which complete unilateral ureteric obstruction was induced at 90 days when a few layers of glomeruli had formed. Up-regulation of TGF-beta1 mRNA and protein was observed in obstructed kidneys, compared to sham-operated control organs, after only 10 days. Increased levels of TGF-beta1 receptors I (TGF-betaR1) and II (TGF-betaR2) were also detected on Western blot, and the cytokine and TGF-betaR1 co-localized in disrupted epithelia on immunohistochemistry. De novo expression of alpha smooth muscle actin, a structural protein up-regulated during TGF-beta1-induced phenotypic switching between human renal dysplastic epithelial and mesenchymal lineages in vitro, was also observed in these aberrant epithelia. These findings implicate increased TGF-beta1 signaling in the early biological changes generated by fetal urinary tract obstruction. PMID- 11438460 TI - High-resolution chromosome 3p allelotyping of breast carcinomas and precursor lesions demonstrates frequent loss of heterozygosity and a discontinuous pattern of allele loss. AB - We performed high-resolution allelotyping for loss of heterozygosity (LOH) analysis on microdissected samples from 45 primary breast cancers, 47 mammary preneoplastic epithelial foci, and 18 breast cancer cell lines, using a panel of 27 polymorphic chromosome 3p markers. Allele loss in some regions of chromosome 3p was detected in 39 of 45 (87%) primary breast tumors. The 3p21.3 region had the highest frequency of LOH (69%), followed by 3p22-24 (61%), 3p21.2-21.3 (58%), 3p25 (48%), 3p14.2 (45%), 3p14.3 (41%), and 3p12 (35%). Analysis of all of the data revealed at least nine discrete intervals showing frequent allele loss: D3S1511-D3S1284 (U2020/DUTT1 region centered on D3S1274 with a homozygous deletion), D3S1300-D3S1234 [fragile histidine triad (FHIT)/FRA3B region centered on D3S1300 with a homozygous deletion], D3S1076-D3S1573, D3S4624/Luca2.1 D3S4597/P1.5, D3S1478-D3S1029, D3S1029 (with a homozygous deletion), D3S1612 D3S1537, D3S1293-D3S1597, and D3S1597-telomere; it is more than likely that additional localized regions of LOH not examined in this study also exist on chromosome 3p. In multiple cases, there was discontinuous allele loss at several 3p sites in the same tumor. Twenty-one of 47 (45%) preneoplastic lesions demonstrated 3p LOH, including 12 of 13 (92%) ductal carcinoma in situ, 2 of 7 (29%) apocrine metaplasia, and 7 of 25 (28%) usual epithelial hyperplasia. The 3p21.3 region had the highest frequency of LOH in preneoplastic breast epithelium (36%), followed by 3p21.2-21.3 (20%), 3p14.2/FHIT region (11%), 3p25 (10%), and 3p22-24 (5%). In 39 3p loci showing LOH in both the tumor and accompanying preneoplasia, 34 (87%) showed loss of the same parental allele (P = 1.2 x 10(-6), cumulative binomial test). In addition, when 21 preneoplastic samples showing LOH were compared to their accompanying cancers, 67% were clonally related, 20% were potentially clonally related but were divergent, and 13% were clonally unrelated. Overall this demonstrated the high likelihood of clonal relatedness of the preneoplastic foci to the tumors. We conclude that: chromosome 3p allele loss is a common event in breast carcinoma pathogenesis; involves multiple, localized sites that often show discontinuous LOH with intervening markers retaining heterozygosity; and is seen in early preneoplastic stages, which demonstrate clonal relatedness to the invasive cancer. PMID- 11438461 TI - Morphological interactions of interdigitating dendritic cells with B and T cells in human mesenteric lymph nodes. AB - Interdigitating dendritic cells (IDC) of the human mesenteric lymph nodes (LN) were examined by two-color immunofluorescent microscopy and flow cytometry to clarify their exact localization, immunophenotype, and relationships with T and B cells. IDC were identified as HLA-DR(bright) large dendriform cells of the T cell areas co-expressing CD40, CD54 (ICAM-1), CD80 (B7/B7-1), CD83, and CD86 (B70/B7 2). The majority of IDC directly attached to a few IgD+ naive B cells as well as to numerous CD4+ T cells. When LN cells were singly suspended and briefly incubated in vitro, IDC formed clusters with IgD+ IgM+ naive B cells, but not with IgA+ or IgG+ B cells. When suspended LN cells were cultured, clustered B cells disappeared within 7 days, and on prolonged culture, some IDC developed into extensively dendriform cells forming stable complexes with several or sometimes numerous CD4+ IL-2R+ CD40L+ activated T cells. These findings indicate that resting naive B cells actually interact with IDC directly in T cell areas of human secondary lymphoid tissues. IDC have a non-antigen (Ag)-specific, strong affinity for resting naive B cells, but this affinity is transient and IDC cannot form stable complexes with B cells, although they can form stable complexes with activated T cells. It is suggested that the stable IDC/Ag-activated T cell complexes make it possible to capture and to stimulate rare Ag-specific resting naive B cells with high efficiency. PMID- 11438462 TI - Interleukin-8 is a paracrine inducer of fibroblast growth factor 2, a stromal and epithelial growth factor in benign prostatic hyperplasia. AB - Benign prostatic hyperplasia (BPH) is an extremely common disease of older men in which there is benign overgrowth of the prostatic transition zone, leading to obstruction of urine outflow. Fibroblast growth factor (FGF) 2, a potent growth factor for prostatic stromal and epithelial cells, is increased twofold in BPH and its concentration is correlated with stromal proliferation in this condition. Immunohistochemistry of normal and hyperplastic prostate revealed that FGF2 expressing stromal cells were present in higher numbers near the epithelial acini, implying that epithelial cells may express a factor that induces FGF2 expression by stromal cells. Conditioned medium from primary cultures of prostatic epithelial cells was capable of inducing increased expression of FGF2 by primary stromal cultures. Blocking experiments with neutralizing anti interleukin (IL)-8 antibodies and pretreatment with lipopolysaccharide, which down-regulates the IL-8 receptor, show that this inducing activity is because of the presence of IL-8 in the epithelial-conditioned medium. Analysis of normal prostatic peripheral zone and BPH tissue by enzyme-linked immunosorbent assay reveals that IL-8 is present at increased levels in hyperplastic prostate. Therefore IL-8 produced by prostatic epithelial cells can induce FGF2, a potent stromal and epithelial growth factor, and in this manner promote the abnormal proliferation of the prostatic transition zone that is critical in the pathogenesis of BPH. PMID- 11438463 TI - Interleukin-1 induces pro-mineralizing activity of cartilage tissue transglutaminase and factor XIIIa. AB - Two transglutaminases (TGases), Factor XIIIa and tissue TGase (tTGase), are expressed in temporal-spatial association with matrix calcification in growth plates. Meniscal and articular cartilage matrix calcification are prevalent in osteoarthritis (OA) and aging. Here, we demonstrated up-regulation of tTGase and Factor XIIIa in superficial and deep zones of knee OA articular cartilage and the central (chondrocytic) zone of OA menisci. Transforming growth factor-beta and interleukin (IL)-1beta induced Factor XIIIa and tTGase expression in cartilage and meniscal organ cultures. Thus, we studied TGase activity. Donor age dependent, OA severity-related, and IL-1-induced increases in TGase activity were demonstrated in both knee menisci and cultured meniscal cells. Meniscal cell TGase activity was stimulated by nitric oxide donors and tumor necrosis factor alpha, but transforming growth factor-beta did not stimulate TGase activity. The iNOS inhibitor N-monomethylarginine (NMMA) and an inhibitor of tumor necrosis factor receptor-associated factor (TRAF)2 and TRAF6 signaling (the zinc finger protein A20) suppressed IL-1 induction of TGase activity. Increased Factor XIIIa and tTGase activities, achieved via direct transfection of chondrocytic TC28 and meniscal cells, both induced matrix apatite deposition. Thus, Factor XIIIa and tTGase activities were increased in aging, degenerative cartilages and induced by IL-1. Because TGase activity promoted apatite deposition, our findings potentially implicate inflammation in the pathogenesis of cartilage matrix calcification. PMID- 11438464 TI - Controlling tumor-derived and vascular endothelial cell growth: role of the 4Ff2 cell surface antigen. AB - We have isolated a monoclonal antibody, clone betaE11, which recognizes an antigen that is highly abundant on the surface of mitotic vascular endothelial cells and tumor cells. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, expression of this 190-kd antigen is approximately threefold higher in mitotic versus interphase endothelial cells. Treatment of tumor cells with an antibody to the betaE11 antigen inhibits their growth in a dose-dependent manner in vitro with maximal inhibition at an antibody concentration of 1 microg/ml. Different tumor cell lines demonstrate varying sensitivities to anti-betaE11 with the following order of growth inhibition: colon > prostate = glioma > melanoma = fibroblast > breast > liver. Furthermore, the betaE11 antigen localizes to regions of prostatic intraductal neoplasia in paraffin-embedded sections. Mass spectrometry of the cell-derived betaE11 protein and V8-protease fingerprint analysis indicate that the betaE11 antigen is nearly identical to the 4F2 heavy chain antigen, a cell surface protein that has been implicated in cell activation and proliferation. Expression of the betaE11 antigen during mitosis functionally links it to a fundamental aspect of cell proliferation, and its spatial localization on the surface of both proliferating endothelium and tumor cells demonstrates its potential for tumor immunotherapy. PMID- 11438465 TI - Primary renal neoplasms with the ASPL-TFE3 gene fusion of alveolar soft part sarcoma: a distinctive tumor entity previously included among renal cell carcinomas of children and adolescents. AB - The unbalanced translocation, der(17)t(X;17)(p11.2;q25), is characteristic of alveolar soft part sarcoma (ASPS). We have recently shown that this translocation fuses the TFE3 transcription factor gene at Xp11.2 to ASPL, a novel gene at 17q25. We describe herein eight morphologically distinctive renal tumors occurring in young people that bear the identical ASPL-TFE3 fusion transcript as ASPS, with the distinction that the t(X;17) translocation is cytogenetically balanced in these renal tumors. A relationship between these renal tumors and ASPS was initially suggested by the cytogenetic finding of a balanced t(X;17)(p11.2;q25) in two of the cases, and the ASPL-TFE3 fusion transcripts were then confirmed by reverse transcriptase-polymerase chain reaction. The morphology of these eight ASPL-TFE3 fusion-positive renal tumors, although overlapping in some aspects that of classic ASPS, more closely resembles renal cell carcinoma (RCC), which was the a priori diagnosis in all cases. These tumors demonstrate nested and pseudopapillary patterns of growth, psammomatous calcifications, and epithelioid cells with abundant clear cytoplasm and well-defined cell borders. By immunohistochemistry, four tumors were negative for all epithelial markers tested, whereas four were focally positive for cytokeratin and two were reactive for epithelial membrane antigen (EMA) (one diffusely, one focally). Electron microscopy of six tumors demonstrated a combination of ASPS-like features (dense granules in four cases, rhomboid crystals in two cases) and epithelial features (cell junctions in six cases, microvilli and true glandular lumens in three cases). Overall, although seven of eight tumors demonstrated at least focal epithelial features by electron microscopy or immunohistochemistry, the degree and extent of epithelial differentiation was notably less than expected for typical RCC. We confirmed the balanced nature of the t(X;17) translocation by fluorescence in situ hybridization in all seven renal tumors thus analyzed, which contrasts sharply with the unbalanced nature of the translocation in ASPS. In summary, a subset of tumors previously considered to be RCC in young people are in fact genetically related to ASPS, although their distinctive morphological and genetic features justify their classification as a distinctive neoplastic entity. Finally, the finding of distinctive tumors being associated with balanced and unbalanced forms of the same translocation is to our knowledge, unprecedented. PMID- 11438466 TI - Experimental autoimmune myocarditis in A/J mice is an interleukin-4-dependent disease with a Th2 phenotype. AB - Myocarditis in humans is often associated with an autoimmune process in which cardiac myosin (CM) is a major autoantigen. Experimental autoimmune myocarditis (EAM) is induced in mice by immunization with CM. We found that EAM in A/J mice exhibits a Th2-like phenotype demonstrated by the histological picture of the heart lesions (eosinophils and giant cells) and by the humoral response (association of IgG1 response with disease and up-regulation of total IgE). Blocking interleukin (IL)-4 with anti-IL-4 monoclonal antibody (mAb) reduced the severity of EAM. This reduction in severity was associated with a shift from a Th2-like to a Th1-like phenotype represented by a reduction in CM-specific IgG1; an increase in CM-specific IgG2a; an abrogation of total IgE response; a decrease in IL-4, IL-5, and IL-13; as well as a dramatic increase in interferon (IFN) gamma production in vitro. Based on the latter finding, we hypothesized that IFN gamma limits disease. Indeed, IFN-gamma blockade with a mAb exacerbated disease. The ameliorating effect of IL-4 blockade was abrogated by co-administration of anti-IFN-gamma mAb. Thus, EAM represents a model of an organ-specific autoimmune disease associated with a Th2 phenotype, in which IL-4 promotes the disease and IFN-gamma limits it. Suppression of IFN-gamma represents at least one of the mechanisms by which IL-4 promotes EAM. PMID- 11438467 TI - Overall survival in aggressive B-cell lymphomas is dependent on the accumulation of alterations in p53, p16, and p27. AB - Different studies have already shown that the isolated inactivation of p21, p16, or p27 cyclin-dependent kinase inhibitors (CKIs) is associated with increased growth fraction, tumor progression, or decreased overall survival in cases of non Hodgkin's lymphoma. In this study we linked molecular study of the p53 and p16 genes with immunohistochemical analysis of p27 expression in a group of aggressive B-cell lymphomas [large B-cell lymphomas (LBCLs) and Burkitt's lymphomas]. This was done to analyze the relationship between p53 and p16 silencing, p27 anomalous overexpression, and clinical follow-up, testing the hypothesis that the accumulation of CKI alterations could confer to the tumors a higher aggressivity. In a group of 62 patients, p53 inactivation as a result of mutation was observed in 11 cases (18%) and p16 silencing was seen in 27 cases (43.5%) as a result of methylation (20 of 62), 9p21 deletion (7 of 44), or p16 mutation (2 of 62). The simultaneous inactivation of p53 and p16 was detected exclusively in five LBCL cases. Anomalous expression of p27, which has been proven to be associated with the absence of p27/CDK2 complexes and the formation of p27/cyclin D3 complexes where p27 is inactivated, was detected in 19 of 61 cases (31%). Cases characterized by p27 anomalous expression display concurrent inactivation of p21 (provided by p53 mutations) and/or p16 CKIs in 11 of 14 LBCL cases (P = 0.040). When the relationship between the association of inactivated CKIs and overall survival was considered, a significant relationship was found between a lower overall survival probability and an increased number of inactivated CKIs in LBCL cases, with the worst prognosis for the cases displaying concurrent p53, p16, and p27 alterations. This proves that simultaneous inactivation of different tumor suppressor pathways does indeed take place, and that tumor aggressiveness takes advantage of this CKI-concerted silencing. In this same series of data, Burkitt's lymphoma patients seem to behave in a different way than LBCLs, with p53 and p16 alteration being mutually exclusive and the association with p27 anomalous expression not being clinically significant. These facts seem to support that the additive effect of the inactivation of different CKIs could be dependent of the histological type. PMID- 11438468 TI - High expression of the trefoil protein TFF1 in interval breast cancers. AB - Breast cancer screening is important for the early detection of breast cancer. Tumors that become symptomatic in the screening interval are known as interval cancers but the reasons for their rapid progression are unknown. Estrogen receptor expression is lower in interval cancers suggesting that they may have reduced hormonal responsiveness. To investigate this hypothesis we have measured the expression of the estrogen receptor and three estrogen-responsive genes (cathepsin D, progesterone receptor, and TFF1) in screen-detected and interval breast cancers. The expression of the protease cathepsin D was not associated with estrogen receptor in either group of tumor. Progesterone receptor expression was highly correlated with that of the estrogen receptor in both groups of tumors but it was not expressed at significantly different levels in the two groups of tumors. Expression of TFF1, a cellular motogen, was correlated with estrogen receptor in screen-detected but not interval cancers and was expressed at markedly higher levels in interval breast tumors, the group that expresses lower levels of estrogen receptor. Interval cancers are characterized by high levels of expression of TFF1 and/or Ki67 suggesting that cell migration and cell division play important roles in the rapid progression of interval cancers. The observation that TFF1 expression in interval cancers tends to be estrogen independent and that interval cancers have reduced estrogen receptor expression suggests they may have a reduced response to hormone therapy. PMID- 11438469 TI - Expresson of vascular endothelial growth factor, its receptors (FLT-1, KDR) and TSP-1 related to microvessel density and patient outcome in vertical growth phase melanomas. AB - Microvessel density (MVD) was estimated in a series of 202 vertical growth phase (VPG) melanomas and 68 corresponding metastases, using a marker for angiogenic endothelial cells (CD105) and Factor-VIII. The expression pattern of vascular endothelial growth factor (VEGF), FLT-1, KDR and thrombospondin-1 (TSP-1) was studied by immunohistochemistry, in situ hybridization and reverse-transcriptase polymerase chain reaction. CD105 stained significantly less vessels, but gave only limited additional prognostic information compared with Factor-VIII, and MVD was an independent prognostic factor for both markers. Ninety-eight percent of all cases showed expression of VEGF, and higher expression was found significantly more frequent in thinner and less vascularized tumors. Possible autocrine loops were suggested by co-expression of VEGF and its two receptors in tumor cells, and by a significant correlation between KDR and tumor cell proliferation (Ki-67) in the subgroup of thicker tumors. Staining of VEGF receptors in endothelium was not correlated with MVD. Strong expression of TSP-1 in tumor stroma was found in 43% of the primary tumors, and was significantly correlated with increased thickness, proliferation and MVD, as well as decreased survival. These data suggest that MVD is associated with prognosis in cutaneous melanomas, and that the VEGF system and particularly TSP-1 seem to be involved in the regulation of angiogenesis and progression of these tumors. PMID- 11438470 TI - Plasmacytoid dendritic cells (natural interferon- alpha/beta-producing cells) accumulate in cutaneous lupus erythematosus lesions. AB - Plasmacytoid dendritic cell (P-DC) precursors in peripheral blood produce large amounts of interferon (IFN)-alpha/beta when triggered by viruses. However, when incubated with interleukin-3 and CD40 ligand, the same precursors differentiate into mature DCs that stimulate naive CD4(+) T cells to produce Th2 cytokines. We recently reported that P-DCs accumulate in nasal mucosa of experimentally induced allergic rhinitis, supporting a role for this DC subset in Th2-dominated inflammation. Here we examined whether P-DCs accumulate in cutaneous lesions of lupus erythematosus (LE), a disorder associated with increased IFN-alpha/beta production. Our results showed that P-DCs were present in 14 out of 15 tissue specimens of cutaneous LE lesions, but not in normal skin. Importantly, the density of P-DCs in affected skin correlated well (r(s) = 0.79,P < 0.0005) with the high number of cells expressing the IFN-alpha/beta-inducible protein MxA, suggesting that P-DCs produce IFN-alpha/beta locally. Accumulation of P-DCs coincided also with the expression of L-selectin ligand peripheral lymph node addressin on dermal vascular endothelium, adding further support to the notion that these adhesion molecules are important in P-DC extravasation to peripheral tissue sites. Together, our findings suggested that P-DCs are an important source of IFN-alpha/beta in cutaneous LE lesions and may therefore be of pathogenic importance. PMID- 11438471 TI - Selective thrombosis of tumor blood vessels in mammary adenocarcinoma implants in rats. AB - Adenocarcinomas in rats and humans frequently contain perivascular, degranulating mast cells that release heparin. Protamine is a low-molecular weight, cationic polypeptide that binds avidly to heparin and neutralizes its anticoagulant properties. We hypothesized that mast-cell heparin functions as a localized anticoagulant that modulates hemostasis and blood perfusion in tumors. Consequently, systemically administered protamine should be able to neutralize the endogenous heparin within tumors, thereby inducing selective thrombosis of blood vessels within tumors. Here we demonstrate with magnetic resonance imaging (MRI) that an intravenous dose of protamine labeled with gadolinium accumulated within the parenchyma of subcutaneous implants of a mammary adenocarcinoma in Fischer 344 rats. Moreover, we show with dynamic contrast enhanced MRI that sequential intravenous doses of protamine in 12 tumor-bearing rats resulted in significantly decreased signal enhancement kinetics (blood perfusion) of the tumor. This functional impairment of MRI signal enhancement was accompanied by histological evidence of thrombosis in the blood vessels within the tumor. There was no histological evidence of thrombosis within normal liver, kidney, lung, spleen, or adjacent muscle of tumor-bearing animals that received protamine treatment or in the tumors of animals that had not been pretreated with protamine. On the basis of these results, we conclude that protamine accumulates within adenocarcinoma implants and induces selective thrombosis of blood vessels within the tumor, probably by neutralizing the endogenous heparin within tumors. PMID- 11438472 TI - Immunoglobulin gene mutations and frequent use of VH1-69 and VH4-34 segments in hepatitis C virus-positive and hepatitis C virus-negative nodal marginal zone B cell lymphoma. AB - Nodal marginal zone B-cell lymphoma (NMZL) is actually considered as a distinct entity that must be distinguished from extra-nodal and splenic marginal zone lymphomas. To define the cell origin and the role of antigen stimulation we determined the nucleotide sequence of the tumor-related immunoglobulin heavy chain variable genes in 10 cases of NMZL. The results were also evaluated on the basis of the presence of chronic hepatitis C virus (HCV) infection. All 10 cases harbored VH somatic mutations with a sequence homology compared to the closest germline gene, ranging from 83.33 to 98.28%. Interestingly, different VH segments were preferentially used in HCV-positive and HCV-negative patients: three of five HCV-negative NMZLs used a VH4-34 segment joined with different D and JH segments whereas three of five HCV-positive NMZLs used a VH1-69 gene joined with a D3-22 and a JH4 segment, with very strong similarities in the CDR3s among the three different cases. These data indicate: 1) NMZL is derived from B cells that have experienced the germinal center reaction; 2) the preferential usage of a VH1-69 segment in the majority of the HCV-positive NMZL cases with similar CDR3s suggests the presence of a common antigen, probably a HCV antigen epitope, involved in the B-cell selection; and 3) the use of a VH4-34 segment suggests a role of yet unknown B-cell superantigen(s) in the selection of tumor B-cell precursors in HCV-negative NMZL. PMID- 11438473 TI - Alteration of transforming growth factor-beta1 response involves down-regulation of Smad3 signaling in myofibroblasts from skin fibrosis. AB - Fibrosis is an unregulated tissue repair process whose predominant characteristics are the proliferation of myofibroblasts and an excessive deposition of extracellular matrix. Transforming growth factor (TGF)-beta1 is considered as one of the most fibrogenic cytokines. However, the molecular mechanisms involved in its profibrotic role are not fully understood. Here, we addressed the role of TGF-beta1 on cell proliferation and intracellular signal transduction in a pig model of skin fibrosis induced by gamma-irradiation. Primary myofibroblasts were isolated from the fibrotic tissue and their response to TGF-beta1 was compared to that of normal skin fibroblasts. The present results show that the differentiation of myofibroblasts involves a lack of TGF-beta1 growth inhibition and an impaired TGF-beta1 signaling. Receptor activity and Smad2/4 or Smad3/4 complex formation were similar in both cell types after TGF beta1 treatment. However, the translocation of Smad3 protein into the nucleus was reduced in myofibroblasts as compared to that in fibroblasts, as well as its binding to target DNA sequences and the activation of the Smad binding elements found in the PAI-1. Interestingly, Smad2 was translocated similarly to the nucleus in both cell types suggesting that this protein may function normally in myofibroblasts. We propose that uncoupling of antiproliferative and profibrotic actions of TGF-beta1 in fibrosis may occur through differential regulation of the activities of Smad2 and Smad3 transcription factors. PMID- 11438474 TI - Mechanisms of beta-cell death in response to double-stranded (ds) RNA and interferon-gamma: dsRNA-dependent protein kinase apoptosis and nitric oxide dependent necrosis. AB - Viral infection is one environmental factor that has been implicated as a precipitating event that may initiate beta-cell damage during the development of diabetes. This study examines the mechanisms by which the viral replicative intermediate, double-stranded (ds) RNA impairs beta-cell function and induces beta-cell death. The synthetic dsRNA molecule polyinosinic-polycytidylic acid (poly IC) stimulates beta-cell DNA damage and apoptosis without impairing islet secretory function. In contrast, the combination of poly IC and interferon (IFN) gamma stimulates DNA damage, apoptosis, and necrosis of islet cells, and this damage is associated with the inhibition of glucose-stimulated insulin secretion. Nitric oxide mediates the inhibitory and destructive actions of poly IC + IFN gamma on insulin secretion and islet cell necrosis. Inhibitors of nitric oxide synthase, aminoguanidine, and N(G)-monomethyl-L-arginine, attenuate poly IC + IFN gamma-induced DNA damage to levels observed in response to poly IC alone, prevent islet cell necrosis, and prevent the inhibitory actions on glucose-stimulated insulin secretion. N(G)-monomethyl-L-arginine fails to prevent poly IC- and poly IC + IFN-gamma-induced islet cell apoptosis. PKR, the dsRNA-dependent protein kinase that mediates the antiviral response in infected cells, is required for poly IC- and poly IC + IFN-gamma-induced islet cell apoptosis, but not nitric oxide-mediated islet cell necrosis. Alone, poly IC fails to stimulate DNA damage in islets isolated from PKR-deficient mice; however, nitric oxide-dependent DNA damage induced by the combination of poly IC + IFN-gamma is not attenuated by the genetic absence of PKR. These findings indicate that dsRNA stimulates PKR dependent islet cell apoptosis, an event that is associated with normal islet secretory function. In contrast, poly IC + IFN-gamma-induced inhibition of glucose-stimulated insulin secretion and islet cell necrosis are events that are mediated by islet production of nitric oxide. These findings suggest that at least one IFN-gamma-induced antiviral response (islet cell necrosis) is mediated through a PKR-independent pathway. PMID- 11438475 TI - Phenotypic characterization of five dendritic cell subsets in human tonsils. AB - Heterogeneous expression of several antigens on the three currently defined tonsil dendritic cell (DC) subsets encouraged us to re-examine tonsil DCs using a new method that minimized DC differentiation and activation during their preparation. Three-color flow cytometry and dual-color immunohistology was used in conjunction with an extensive panel of antibodies to relevant DC-related antigens to analyze lin(-) HLA-DR(+) tonsil DCs. Here we identify, quantify, and locate five tonsil DC subsets based on their relative expression of the HLA-DR, CD11c, CD13, and CD123 antigens. In situ localization identified four of these DC subsets as distinct interdigitating DC populations. These included three new interdigitating DC subsets defined as HLA-DR(hi) CD11c(+) DCs, HLA-DR(mod) CD11c(+) CD13(+) DCs, and HLA-DR(mod) CD11c(-) CD123(-) DCs, as well as the plasmacytoid DCs (HLA-DR(mod) CD11c(-) CD123(+)). These subsets differed in their expression of DC-associated differentiation/activation antigens and co-stimulator molecules including CD83, CMRF-44, CMRF-56, 2-7, CD86, and 4-1BB ligand. The fifth HLA-DR(mod) CD11c(+) DC subset was identified as germinal center DCs, but contrary to previous reports they are redefined as lacking the CD13 antigen. The definition and extensive phenotypic analysis of these five DC subsets in human tonsil extends our understanding of the complexity of DC biology. PMID- 11438476 TI - Microsatellite instability and high content of activated cytotoxic lymphocytes identify colon cancer patients with a favorable prognosis. AB - Colorectal cancers with high-frequency microsatellite instability show peculiar clinicopathological features and a favorable clinical outcome. We investigated whether the improved prognosis for these cancers is related to the content of activated cytotoxic intraepithelial T lymphocytes. Microsatellite instability and the amount of activated cytotoxic lymphocytes were analyzed according to clinicopathological features, survival, and disease recurrence in 109 right-sided colon carcinomas from 245 consecutive patients with stage II/III colon cancer that underwent radical surgery. High-frequency microsatellite instability was found in 43% of stage II/III proximal colon cancers and was associated with significantly higher numbers of activated cytotoxic lymphocytes. High-frequency microsatellite instability, as well as the content of intratumoral-activated cytotoxic T lymphocytes correlated with improved overall and disease-free survival, particularly in patients with stage III tumors. Multivariate analysis revealed that patients with both features had a risk of death and relapse markedly lower than that associated with microsatellite status or intratumoral cytotoxic lymphocytes separately. The presence of local cytotoxic immune responses is probably the major determinant of the good clinical course of patients with microsatellite unstable colon cancer. Furthermore, high-frequency microsatellite instability coupled with a high content of intratumoral cytotoxic lymphocytes may identify a subset of colon cancer patients with a favorable clinical outcome, particularly in stage III disease. PMID- 11438477 TI - Mitochondrial release of apoptosis-inducing factor and cytochrome c during smooth muscle cell apoptosis. AB - Photodynamic therapy (PDT) is under investigation for the treatment of intimal hyperplastia in conditions such as atherosclerosis and restenosis. Although smooth muscle cells (SMCs) may be a key target for treatment, the effects of PDT on these cells are poorly characterized. In the present study, apoptosis was induced in primary human aortic SMCs by the combination of the photosensitizer verteporfin and visible light. After PDT, an increase in mitochondrial cytochrome c (cyt c) and apoptosis-inducing factor (AIF) levels were detected in the cytosol immediately and their levels increased steadily up to 2 hours. Cytosolic levels of the pro-apoptotic Bcl-2 family member Bax decreased reciprocally throughout this period, but this change did not occur before cyt c release. Confocal microscopy revealed a diffuse staining pattern of cyt c within apoptotic cells as compared to a distinct mitochondrial staining in normal cells. AIF translocated from mitochondria to the nucleus during the progression of apoptosis. After cyt c release, caspase-9 and caspase-3 processing was visible by 1 hour and caspase-6, 7, and -8 processing was apparent by 2 hours after PDT. In summary, these results demonstrate for the first time the cellular redistribution of mitochondrial AIF during SMC apoptosis, as well as the early release of cyt c and the subsequent activation of multiple caspases during PDT-induced SMC apoptosis. PMID- 11438478 TI - Inhibition of choroidal neovascularization by intravenous injection of adenoviral vectors expressing secretable endostatin. AB - Endostatin is a cleavage product of collagen XVIII that inhibits tumor angiogenesis and growth. Interferon alpha2a blocks tumor angiogenesis and causes regression of hemangiomas, but has no effect on choroidal neovascularization (CNV). Therefore, inhibitors of tumor angiogenesis do not necessarily inhibit ocular neovascularization. In this study, we used an intravenous injection of adenoviral vectors containing a sig-mEndo transgene consisting of murine immunoglobulin kappa-chain leader sequence coupled to sequence coding for murine endostatin to investigate the effect of high serum levels of endostatin on CNV in mice. Mice injected with a construct in which sig-mEndo expression was driven by the Rous sarcoma virus promoter had moderately high serum levels of endostatin and significantly smaller CNV lesions at sites of laser-induced rupture of Bruch's membrane than mice injected with null vector. Mice injected with a construct in which sig-mEndo was driven by the simian cytomegalovirus promoter had approximately 10-fold higher endostatin serum levels and had nearly complete prevention of CNV. There was a strong inverse correlation between endostatin serum level and area of CNV. This study provides proof of principle that gene therapy to increase levels of endostatin can prevent the development of CNV and may provide a new treatment for the leading cause of severe loss of vision in patients with age-related macular degeneration. PMID- 11438479 TI - Matrix metalloproteinase-2 is associated with tenascin-C in calcific aortic stenosis. AB - We previously showed that the expression of tenascin (TN-C), an extracellular matrix glycoprotein found in developing bone and atherosclerotic plaque, and matrix metalloproteinase-2 (MMP-2) are coordinated and interdependent in cultured vascular smooth muscle cells. In this study, we hypothesized that TN-C and MMP-2 are mechanistically involved in the pathobiology of calcific aortic stenosis. Human calcific aortic stenosis cusps demonstrated immunohistochemically prominent deposition of TN-C, MMP-2, and alkaline phosphatase activity, as well as MMP-2 gelatinolytic activity. Although far lesser amounts of TN-C were noted in several of the grossly non-calcified valve cusps, MMP-2 and AP were never detected. Further, when aortic valve interstitial cells (both sheep and human) were cultivated on collagen supplemented with TN-C, both MMP-2 mRNA expression and MMP 2 gelatinolytic activity (both pro and active forms), were up-regulated compared to control. These observations support the view that accumulation of first TN-C and then MMP-2 are associated with progression of calcification. The residual presence of these proteins in severe calcifications is indicative of their involvement in the pathogenesis. PMID- 11438480 TI - Ethanol-induced apoptosis in mouse liver: Fas- and cytochrome c-mediated caspase 3 activation pathway. AB - Hepatic apoptosis has been shown to occur in both experimental and clinical alcoholic liver disease, but the signaling pathway remains unknown. This study was undertaken to examine specifically the involvement of the upstream signals, Fas and cytochrome c, in alcohol-induced caspase-3 activation and apoptosis in the liver. Male FVB mice were administrated intragastrically a single dose of alcohol at 6 g/kg, which has been shown to represent binge drinking in humans. Hepatic apoptosis was detected by a terminal deoxynucleotidyl transferase dUTP nick-end labeling assay. Active form of caspase-3 was identified by immunoperoxidase staining and confirmed by immunogold labeling and was found to be in the cytosol and nucleus. Enzymic assay further confirmed caspase-3 activation and nucleus localization. Systemic administration of caspase-3 inhibitor, Ac-DEVD-FMK, inhibited caspase-3 activity and abrogated apoptosis. Elevation of cytosolic cytochrome c was found by immunoperoxidase staining, immunogold labeling, and Western blot. Increased Fas ligand expression was detected by immunoperoxidase staining. Intravenous administration of a neutralizing Fas ligand monoclonal antibody resulted in suppression of caspase-3 activation and attenuation of apoptosis, but did not inhibit mitochondrial cytochrome c release. The results thus demonstrate that Fas/Fas ligand system mediated caspase-3 activation plays a central role in the ethanol-induced hepatic apoptosis. PMID- 11438481 TI - A close association of torsinA and alpha-synuclein in Lewy bodies: a fluorescence resonance energy transfer study. AB - TorsinA, a novel protein in which a mutation causes dominant, early onset torsion dystonia, may serve as a chaperone for misfolded proteins that require refolding or degradation. It has been hypothesized that misfolded alpha-synuclein, a protein in which two mutations cause autosomal dominantly inherited Parkinson's disease, serves as a nidus for the development of a Lewy body. We hypothesized that torsinA plays a role in the cellular processing of alpha-synuclein. We demonstrate that anti-torsin antibodies stain Lewy bodies and Lewy neurites in the substantia nigra and cortex. Using sensitive fluorescent resonance energy transfer (FRET) techniques, we find evidence of a close association between torsinA and alpha-synuclein in Lewy bodies. PMID- 11438482 TI - PGF(2alpha), a prostanoid released by endothelial cells activated by hypoxia, is a chemoattractant candidate for neutrophil recruitment. AB - Despite increasing evidence supporting the involvement of neutrophils in ischemic and postischemic damages, the mechanisms underlying the early recruitment of these cells are not completely understood. In this report, the effects of conditioned media from hypoxic endothelial cells on neutrophil chemotaxis were investigated by biochemical and morphological studies. We showed that conditioned media collected from several endothelial cell origins submitted to hypoxia as well as ischemic rat liver perfusion liquids have a chemotactic activity for neutrophils. The role of various chemoattractant molecules like HETEs, platelet activating factor, and cytokines such as interleukin-8 and interleukin-1 was examined in the same model. Chemotactic peptide contribution was ruled out as boiled conditioned media still trigger chemotaxis. However, cell treatment with cyclooxygenase inhibitors, neutralization of PGF(2alpha) biological activity with polyclonal antibodies, and the neutrophil preincubation with a specific PGF(2alpha) antagonist, all dramatically inhibited neutrophil chemotaxis. A strong chemoattractant effect of pure exogenous PGF(2alpha) or of a synthetic analog was also observed. The major effect of PGF(2alpha) on neutrophil chemotaxis was confirmed ex vivo in a rat liver perfusion ischemic model. These results suggest that PGF(2alpha), a prostanoid abundantly released by the endothelium of hypoxic or ischemic tissues, is a chemoattractant molecule that might be involved in the early recruitment of neutrophils in ischemic organs. PMID- 11438483 TI - PTEN is a target of chromosome 10q loss in anaplastic oligodendrogliomas and PTEN alterations are associated with poor prognosis. AB - Allelic loss of 10q is a common genetic event in malignant gliomas, with three 10q tumor suppressor genes, ERCC6, PTEN, and DMBT1, putatively implicated in the most common type of malignant glioma, glioblastoma. Anaplastic oligodendroglioma, another type of malignant glioma, provides a unique opportunity to study the relevance of particular genetic alterations to chemosensitivity and survival. We therefore analyzed these three genes in 72 anaplastic oligodendrogliomas. Deletion mapping demonstrated 10q loss in 14 of 67 informative cases, with the PTEN and DMBT1 regions involved in all deletions but with the ERCC6 locus spared in two cases. Seven tumors had PTEN gene alterations; two had homozygous DMBT1 deletions, but at least one reflected unmasking of a germline DMBT1 deletion. No mutations were found in ERCC6 exon 2. Chemotherapeutic response occurred in two of the seven tumors with PTEN alterations, but with unexpected short survival times. PTEN gene alterations were not associated with poor therapeutic response in multivariate analysis, but were independently predictive of poor prognosis even after multivariate adjustment for both 10q and 1p loss. In anaplastic oligodendroglioma, therefore, PTEN is a target of 10q loss, and PTEN alterations are associated with poor prognosis, even in chemosensitive cases. PMID- 11438484 TI - Transgenic expression of cyclin-dependent kinase 4 results in epidermal hyperplasia, hypertrophy, and severe dermal fibrosis. AB - In a previous report we have described the effects of expression of D-type cyclins in epithelial tissues of transgenic mice. To study the involvement of the D-type cyclin partner cyclin-dependent kinase 4 (CDK4) in epithelial growth and differentiation, transgenic mice were generated carrying the CDK4 gene under the control of a keratin 5 promoter. As expected, transgenic mice showed expression of CDK4 in the epidermal basal-cell layer. Epidermal proliferation increased dramatically and basal cell hyperplasia and hypertrophy were observed. The hyperproliferative phenotype of these transgenic mice was independent of D-type cyclin expression because no overexpression of these proteins was detected. CDK4 and CDK2 kinase activities increased in transgenic animals and were associated with elevated binding of p27(Kip1) to CDK4. Expression of CDK4 in the epidermis results in an increased spinous layer compared with normal epidermis, and a mild hyperkeratosis in the cornified layer. In addition to epidermal changes, severe dermal fibrosis was observed and part of the subcutaneous adipose tissue was replaced by connective tissue. Also, abnormal expression of keratin 6 associated with the hyperproliferative phenotype was observed in transgenic epidermis. This model provides in vivo evidence for the role of CDK4 as a mediator of proliferation in epithelial cells independent of D-type cyclin expression. PMID- 11438485 TI - Are syndromic fundic gland polyps true neoplasms? PMID- 11438486 TI - Plexiform lesion in severe pulmonary hypertension: association with glomeruloid lesion. PMID- 11438487 TI - Sarcomatoid variant of anaplastic large cell lymphoma with cytoplasmic ALK and alpha-smooth muscle actin expression: a mimic of inflammatory myofibroblastic tumor. PMID- 11438489 TI - Esophageal motility in reflux disease before and after fundoplication: a prospective, randomized, clinical, and manometric study. AB - BACKGROUND AND AIMS: The purpose of this study was to determine whether esophageal dysmotility affects symptoms of gastroesophageal reflux disease or clinical outcome after laparoscopic fundoplication and whether esophagus motor function changes postoperatively. METHODS: Two hundred patients with a history of long-standing gastroesophageal reflux disease were investigated by clinical assessment, upper gastrointestinal endoscopy, esophageal manometry, and 24-hour pH monitoring between May 1999 and May 2000. Patients were stratified according to presence or absence of esophageal dysmotility (each n = 100) and randomized to either 360 degrees (Nissen) or 270 degrees (Toupet) fundoplication. At a 4-month postoperative follow-up, preoperative tests were repeated. RESULTS: Preoperative esophageal dysmotility was associated with more severe reflux symptoms, more frequent resistance to medical treatment (64% vs. 49%; P < 0.05), and greater decrease in lower esophageal sphincter pressure (9.5 +/- 5.3 vs. 12.4 +/- 6.7 mm Hg; P < 0.0005) compared with normal motility. Postoperatively, clinical outcome and reflux recurrence (21% vs. 14%) were similar. Esophageal motility remained unchanged in 85% of patients and changed from pathologic to normal in 20 (10 Nissen/10 Toupet) and vice versa in 9 (8 Nissen/1 Toupet) patients. CONCLUSIONS: Esophageal dysmotility (1) reflects more severe disease; (2) does not affect postoperative clinical outcome; (3) is not corrected by fundoplication, independent of the surgical procedure performed; (4) may occur as a result of fundoplication; and (5) requires no tailoring of surgical management. PMID- 11438490 TI - Restricted V(H) gene usage in lamina propria B cells producing anticolon antibody from patients with ulcerative colitis. AB - BACKGROUND AND AIMS: Autoimmune responses against colonic epithelium may play a role in the development of colonic inflammation associated with ulcerative colitis (UC). In this study, we established and characterized B-cell lines and clones that produced anticolon antibody from inflamed colonic mucosa of UC subjects. METHODS: B-cell lines were generated through Epstein-Barr virus transformation of lamina propria lymphocytes (LPLs) from colonic mucosa and peripheral blood lymphocytes, and these lines were screened for the production of anticolon antibodies. B-cell lines were then cloned by limiting dilution culture, and messenger RNA expression of immunoglobulin heavy-chain variable region (V(H)) was assessed. RESULTS: V(H) gene families used in B-cell lines established from LPLs of normal controls were diverse, and B-cell lines from UC LPLs expressed a restricted V(H)3 family usage. All 15 clones from UC used a restricted V(H)3 gene family, whereas diverse V(H) gene families were used by 24 clones from normal controls. The analysis of nucleotide sequences indicated that these clones were derived from various germline gene segments. CONCLUSIONS: The restricted V(H) gene usage in anticolon autoantibodies producing B-cell clones suggests that a particular antigenic stimulus contributes to the pathogenesis of UC. PMID- 11438491 TI - Functional analysis of RET with Hirschsprung mutations affecting its kinase domain. AB - BACKGROUND AND AIMS: Many missense mutations in the RET proto-oncogene were found in familial and sporadic cases of Hirschsprung disease (HSCR). The aim of this study was to make clear the mechanisms of RET dysfunction by HSCR mutations identified in its kinase domain. METHODS: Ten kinase domain HSCR mutations were introduced into wild-type RET and constitutively activated RET with a multiple endocrine neoplasia 2A mutation, and the resulting mutant complementary DNAs were transfected into SK-N-MC primitive neuroectodermal tumor cells or NIH 3T3 fibroblast cells. The levels of activation of mutant RET and representative signaling molecules were investigated in the transfectants. RESULTS: E762Q, S767R, R972G, and M980T mutations partially impaired the RET kinase activity and the representative signaling pathways. In particular, these mutations severely impaired the phospholipase C-gamma signaling pathway in SK-N-MC cells. S765P, R873Q, F893L, R897Q, and E921K mutations resulted in a complete loss of the RET kinase activity. The P973L mutation markedly decreased the expression of the RET protein with normal kinase activity. CONCLUSIONS: Hirschsprung disease can result from these distant functional classes of kinase domain mutation of RET. PMID- 11438492 TI - Lower esophageal sphincter is achalasic in nNOS(-/-) and hypotensive in W/W(v) mutant mice. AB - BACKGROUND AND AIMS: It has been proposed that nitrergic nerves mediate lower esophageal sphincter (LES) relaxation with intramuscular interstitial cells of Cajal (ICC-IM) as an intermediary. Dysfunction of the nitrergic pathway has been shown to cause LES hypertension and impaired relaxation in achalasia. We determined whether mice with neuronal nitric oxide synthase gene disruption (nNOS(-/-)) and W/W(v) mice lacking ICC-IM have achalasia-like LES dysfunction. METHODS: Intraluminal manometry using a customized micro-sized catheter assembly was performed in anesthetized mice. Basal LES pressure and swallow- and vagal evoked LES relaxations were quantified in wild-type, Nomega-nitro-L-arginine methyl ester HCl salt (L-NAME)-treated, nNOS(-/-), and W/W(v) mice. RESULTS: Wild type mouse LES maintained a basal pressure (24 +/- 3 mm Hg; N = 8) and relaxed normally to swallow (87% +/- 3%; N = 8) and vagal stimulation (91% +/- 4% mm Hg; N = 6). Pretreatment with L-NAME (100 mg/kg, intravenously) attenuated LES relaxation to both stimuli (P < 0.05). The LES in nNOS(-/-) was significantly hypertensive (36 +/- 5 mm Hg; N = 10; P < 0.05) with a markedly impaired relaxation (P < 0.05). In contrast, W/W(v) mouse LES was significantly hypotensive (11 +/- 2 mm Hg; N = 6; P < 0.05) with normal relaxation that was blocked by L-NAME. CONCLUSIONS: nNOS(-/-) mice have LES hypertension with impaired relaxation resembling achalasia. In contrast, W/W(v) mice have hypotensive LES with unimpaired relaxation, suggesting that ICC-IM do not play a role in nitrergic neurotransmission. PMID- 11438493 TI - Neuroendocrine-specific and gastrin-dependent expression of a chromogranin A luciferase fusion gene in transgenic mice. AB - BACKGROUND AND AIMS: Chromogranin A (CgA) is a multifunctional acidic protein specifically expressed in neuroendocrine cells. In the stomach, CgA is found predominantly in enterochromaffin-like (ECL) cells, where it is regulated by gastrin. We investigated the ability of a promoter fragment comprising 4.8 kb of 5'-flanking DNA of the mouse CgA (mCgA) gene to direct cell-specific expression as well as gastrin responsiveness in the gastroenteropancreatic neuroendocrine system. METHODS: Two independent lines of mCgA 4.8 kb-luc transgenic mice were created. Transgene expression was assessed by determination of luciferase activity and reverse-transcription polymerase chain reaction analysis of luciferase messenger RNA. Cell specificity of transgene expression was investigated by immunohistochemical analysis. The influence of hypergastrinemia on transgene expression was determined after repeated omeprazole injections. RESULTS: In both transgenic lines, mCgA 4.8 kb-luc expression paralleled the expression pattern of the endogenous CgA gene. ECL cells were identified as the major gastric cell population expressing the transgene. Omeprazole treatment stimulated expression of the transgene and the endogenous CgA gene selectivity in the gastric corpus (3-4-fold). CONCLUSIONS: mCgA 5'-flanking DNA (4.8 kb) contain the major cis-regulatory element(s) required for cell-specific CgA expression in the neuroendocrine system and gastrin-responsiveness in the gastric corpus. Further analysis of the CgA promoter in transgenic studies may elucidate the general molecular mechanisms underlying cell-specific gene expression in the gastroenteropancreatic neuroendocrine system. PMID- 11438494 TI - TGF-alpha inhibits apoptosis of murine gastric pit cells through an NF-kappaB dependent pathway. AB - BACKGROUND AND AIMS: Recently, some growth factors have been shown to play roles not only as growth factors but also as cell-surviving factors. Transforming growth factor (TGF)-alpha is expressed in normal gastric mucosa. In this study, we investigated the cell-surviving effect of TGF-alpha on gastric mucosal cells and its signaling mechanism. METHODS: We used a gastric mucosal cell line, GSM06, and gastric cancer cell line, AGS. Apoptosis was induced by serum depletion or exposure to sodium butyrate. Analysis of apoptosis was performed by DNA ladder assay, measuring the DNA fragmentation ratio (Burton method), and 4',6-diamidino 2-phenylindole staining. RESULTS: TGF-alpha protected gastric mucosal cells against apoptosis induced by serum depletion or sodium butyrate in a dose dependent manner. This antiapoptotic effect of TGF-alpha was blocked by the pretreatment with reagents that can potentially inhibit NF-kappaB activation, whereas neither MEK inhibitor PD098059 nor PI-3-kinase inhibitor wortmannin abolished this effect. Electrophoretic mobility shift assay showed nuclear factor kappaB (NF-kappaB) activation by TGF-alpha stimulation. TGF-alpha also enhanced the expression of Bcl-2 family proteins in an NF-kappaB-dependent manner. CONCLUSIONS: TGF-alpha plays an antiapoptotic role in gastric mucosal cells via the NF-kappaB-dependent pathway. PMID- 11438495 TI - Growth retardation, duodenal lesions, and aberrant ileum architecture in triple null mice lacking EGF, amphiregulin, and TGF-alpha. AB - BACKGROUND AND AIMS: Mice lacking epidermal growth factor (EGF), transforming growth factor alpha, and amphiregulin were used to identify roles for these EGF receptor (EGF-R) ligands in gastrointestinal development and mucosal integrity. METHODS: Gastrointestinal tract development was examined in knockout mice and correlated with expression of EGF-R protein and EGF family members throughout the gut. Crossfostering experiments addressed roles of maternal- and neonatal-derived ligands in pup growth and intestinal development. Cysteamine-induced ulceration in EGF(-/-) mice was used to examine its role in mucosal cytoprotection. RESULTS: Neonatal mice lacking all 3 ligands were growth retarded, even when reared by wild-type dams; conversely, lack of maternal ligands transiently impaired wild type pup growth. Triple null neonates displayed spontaneous duodenal lesions, and ileal villi were truncated and fragile with reduced cellular proliferation in the crypts. However, maturation of digestive enzymes was unaffected. Adult EGF(-/-) mice displayed more severe lesions in response to cysteamine treatment compared with wild-type counterparts, although triple null mice were not more susceptible to dextran sulfate sodium-induced colitis, suggesting a differential role for these ligands in the injury response. CONCLUSIONS: EGF-R ligands are required for development and mucosal maintenance in mouse small intestine. Both maternal and neonatal sources of growth factors are required for optimal pup growth. PMID- 11438496 TI - Leptin is a growth factor for colonic epithelial cells. AB - BACKGROUND AND AIMS: Obesity increases the risk of colon cancer, whereas physical activity reduces the risk. Plasma levels of leptin increase in proportion to the level of obesity and are reduced by physical activity. Leptin acts as a growth factor for several cell types and thus may provide a biological explanation for the observed epidemiological risk factors. The aim of this study was to investigate whether leptin is a growth factor for colonic epithelial cells. METHODS: The presence of the leptin receptor in human colon cancer cell lines was assessed using reverse-transcription polymerase chain reaction and immunoblotting, and its presence in human colonic tissue was assessed by immunohistochemistry. The effects of leptin in vitro on HT29 cells were assessed by assessing p42/44 mitogen-activated protein kinase phosphorylation, thymidine incorporation, and cell numbers and in vivo in C57BL/6 mice by colonic bromodeoxyuridine incorporation. RESULTS: The leptin receptor is expressed in human colon cancer cell lines and human colonic tissue. Stimulation with leptin leads to phosphorylation of p42/44 mitogen-activated protein kinase and increases proliferation in vitro and in vivo. CONCLUSIONS: Leptin is a growth factor in colonic epithelial cells and one that may provide a biological explanation for the observed associations between obesity, physical activity, and colon cancer. PMID- 11438497 TI - Nonalcoholic fatty liver disease: predictors of nonalcoholic steatohepatitis and liver fibrosis in the severely obese. AB - BACKGROUND AND AIMS: Nonalcoholic fatty liver disease (NAFLD) is common in severely obese subjects and can progress to cirrhosis and liver failure. Predicting advanced or progressive disease may help in selecting patients for liver biopsy and assist the development of therapeutic options. METHODS: Liver biopsies were taken at laparoscopic obesity surgery in 105 consecutive patients. The clinical and biochemical variables were analyzed for correlation with specific histologic features. RESULTS: Twenty-six patients (25%) were found to have nonalcoholic steatohepatitis (NASH), and 11 (42%) of these had advanced fibrosis. A raised index of insulin resistance (odds ratio [OR] 9.3, 95% confidence interval [CI] 3.4-26), systemic hypertension (OR 5.2, 95% CI 2.0 13.5), and raised alanine aminotransferase (OR 8.6, 95% CI 3.1-23.5) were independent predictors of NASH. A combination of 2 or 3 of these predictors allows a sensitivity of 0.8 and specificity of 0.89 for NASH. Alcohol consumption was associated with a reduction in NASH (OR 0.35, 95% CI 0.12-1.00) and diabetes (OR 0.18, 95% CI 0.047-0.67). CONCLUSION: Insulin resistance and systemic hypertension, features of the metabolic syndrome, are independently associated with advanced forms of NAFLD. Moderate alcohol consumption seems to reduce the risk of NAFLD in the severely obese, possibly by reducing insulin resistance. PMID- 11438498 TI - Interferon alpha treatment and retreatment of hepatitis B e antigen-negative chronic hepatitis B. AB - BACKGROUND AND AIMS: In hepatitis B e antigen (HBeAg)-negative chronic hepatitis B (CHBe-), interferon (IFN) achieves very low biochemical sustained response rates. No information exists on retreatment. METHODS: Two hundred sixteen CHBe- patients treated for 5 or 12 months with 3 MU IFN-alpha2b thrice weekly and retreated (51 patients, 60 courses) because of no response or relapse were retrospectively analyzed. RESULTS: After 7.0 years of median follow-up, 39 naive patients (18.1%) were still in biochemical and virologic remission after a single IFN course. Longer treatment and a biochemical response within 4 months were significant predictors, inversely related to relapse by multivariate analysis (relative hazard [RH], 0.611; 95% confidence interval [CI], 0.448-0.834 and RH, 0.290; 95% CI, 0.192-0.438, respectively). Retreatment resulted in 18.4% sustained response by intention-to-treat (18 of 98 patients). Patients with sustained response had persistently normal alanine aminotransferase levels, undetectable hepatitis B virus (HBV) DNA by molecular hybridization, and significant improvement of histologic grade, and 32% of them lost hepatitis B surface antigen. In sustained responders, serum HBV DNA was undetectable or very low at the end of treatment and at the end of follow-up (median 3934 and 903 copies/mL, respectively) by quantitative polymerase chain reaction. CONCLUSIONS: IFN induced long-term biochemical and virologic remission in approximately 18% of naive or retreated patients with CHBe-. Sustained responders exhibited significant histologic improvement and a high rate of HBsAg loss. PMID- 11438499 TI - Somatostatin treatment and risk stratification by continuous portal pressure monitoring during acute variceal bleeding. AB - BACKGROUND AND AIMS: During acute variceal bleeding, several factors may lead to elevations of hepatic venous pressure gradient (HVPG), which may precipitate further hemorrhage. Whether somatostatin can suppress these increments is unknown. This study monitored somatostatin effects on HVPG during acute bleeding and assessed whether the changes affect outcome. METHODS: In 40 patients with acute variceal bleeding treated with sclerotherapy, a catheter was placed into a main hepatic vein for 24-hour serial measurements of HVPG. After baseline measurements, patients received somatostatin (N = 25) or placebo (N = 15) under double blind conditions. RESULTS: Somatostatin but not placebo produced a sustained decrease in HVPG (from 20.7 +/- 3.7 mm Hg to 17.7 +/- 2.7, P < 0.01). In patients receiving placebo, HVPG increased after a test meal (P = 0.018) and after blood transfusion (P = 0.034). Somatostatin completely prevented these increments. HVPG decreased significantly only in patients without further bleeding. One of 27 patients with HVPG <20 mm Hg at baseline or decreased >10% rebled vs. 9 of 13 who had neither of these 2 criteria (P < 0.0001). Both criteria had independent prognostic value for further bleeding. CONCLUSIONS: During acute variceal bleeding, somatostatin produces a significant and sustained decrease in HVPG and prevents secondary elevations. Monitoring HVPG may stratify further bleeding risk and discriminate treatment response. PMID- 11438500 TI - Identification of oxysterols in human bile and pigment gallstones. AB - BACKGROUND AND AIMS: Although cholesterol is the most abundant sterol in animal tissues, oxidized products of cholesterol (oxysterols) also occur in mammalian organs and blood and are cytotoxic, atherogenic, and carcinogenic. However, the presence of oxysterols in bile or gallstones has never been reported. METHODS: Fresh human bile and gallstones were collected. Sterol content and structure were analyzed using gas chromatography/mass spectrometry (GC/MS). Bacterial DNA was extracted from human gallstones. RESULTS: GC/MS identified cholesta-4,6-diene-3 one and cholest-4-ene-3-one, with several as yet unidentified oxysterols in bile and stone samples. Several plant and fungal sterols were also present in gallstones. When 102 human gallstones were analyzed for oxysterols, they were markedly higher (as percent of total sterols) in pigment gallstones, where bacterial DNA is most abundant. CONCLUSIONS: These observations suggest biliary oxysterols are associated with the presence of bacteria and may play a role in the pathogenesis of gallstones and biliary tract cancers. PMID- 11438501 TI - A functional polymorphism of the stromelysin gene (MMP-3) influences susceptibility to primary sclerosing cholangitis. AB - BACKGROUND AND AIMS: We have investigated the influence of a biallelic polymorphism of the promoter region of stromelysin (matrix metalloproteinase 3) on susceptibility to primary sclerosing cholangitis (PSC). The 5A allele is associated with increased transcription, compared with wild-type (6A). METHODS: An allelic association study was performed: in stage 1, 52 PSC patients (43 with inflammatory bowel disease [IBD]) and 99 healthy subjects (HS) were genotyped. In stage 2, 59 PSC patients (49 IBD), 84 patients with uncomplicated ulcerative colitis, and 72 HS were genotyped. RESULTS: In stage 1, 5A carriage rate (90.4% vs. 72.7%; P = 0.012) and 5A allelic frequency (65.4% vs. 48.5%; P = 0.005) were increased, and 6A homozygosity was reduced in PSC (9.6% vs. 27.3%; P = 0.012). In stage 2, 5A allelic carriage was increased in PSC (93.2% vs. 76.4% in HS; P = 0.0092) and 6A homozygosity was reduced (6.8% vs. 23.8% in HS; P = 0.0092). Portal hypertension was associated with 5A homozygosity in PSC (P = 0.035; odds ratio [OR], 3.88). In the combined data set, 5A allelic frequencies (63.5% vs. 49.4%; P = 0.001; OR, 1.78) and 5A carriage rates (91.9% vs. 74.2%; P = 0.0002; OR, 3.92) were increased, and 6A homozygosity was reduced in PSC (8.1% vs. 25.7%; P = 0.0002; OR, 0.25). Overall, portal hypertension was associated with 5A homozygosity (P = 0.0192; OR, 3.12). CONCLUSIONS: Stromelysin polymorphism may influence susceptibility and disease progression in PSC. PMID- 11438502 TI - Thrombin-activatable fibrinolysis inhibitor deficiency in cirrhosis is not associated with increased plasma fibrinolysis. AB - BACKGROUND AND AIMS: The bleeding tendency of patients suffering from cirrhosis is in part ascribed to accelerated fibrinolysis. In this study, the role of the recently discovered inhibitor of fibrinolysis, thrombin-activatable fibrinolysis inhibitor (TAFI) in cirrhosis was examined. METHODS: In 64 patients with cirrhosis of varying severity, TAFI antigen levels were measured by enzyme-linked immunosorbent assay and compared with TAFI levels in control subjects. Furthermore, a plasma-based fibrinolysis assay was performed in the presence and absence of a specific inhibitor of activated TAFI. RESULTS: TAFI levels were decreased in cirrhosis. Mean TAFI levels were 66% in Child's A, 55% in Child's B, 47% in Child's C cirrhosis, and 26% in acute liver failure. Decreased TAFI antigen levels were highly correlated with antithrombin and alpha(2)-antiplasmin activity levels. Clot lysis times and clot lysis ratio (defined as ratio between clot lysis time in the absence and presence of a specific inhibitor of activated TAFI) of cirrhotics were not significantly different from healthy controls. CONCLUSIONS: Despite decreased levels of TAFI and other components of the fibrinolytic system, no evidence of increased plasma fibrinolytic potential in cirrhosis is observed using the plasma-based assay of this study. The reduction of antifibrinolytic factors in cirrhosis is compensated by the concomitant reduction in profibrinolytics. PMID- 11438503 TI - The orphan nuclear receptor, shp, mediates bile acid-induced inhibition of the rat bile acid transporter, ntcp. AB - BACKGROUND AND AIMS: Hepatic bile acid homeostasis is regulated by negative feedback inhibition of genes involved in the uptake and synthesis of bile acids. Bile acids down-regulate the rate-limiting gene for bile acid synthesis, cholesterol 7alpha-hydroxylase (cyp7a), via bile acid receptor (fxr) activation of an inhibitory nuclear receptor, shp. We hypothesized that shp would also mediate negative feedback regulation of ntcp, the principal hepatic bile acid transporter. METHODS: Primary rat hepatocytes or transfected HepG2 and Cos cells were treated with retinoids with or without bile acids, and effects on bile acid transport and ntcp and shp gene expression and promoter activity were determined. Gel shift assays were performed using synthetic fxr, rxr, and rar proteins. RESULTS: Bile acid treatment of primary rat hepatocytes prevented retinoid activation of ntcp gene expression and function; this corresponded temporally with shp gene activation. Bile acid-mediated down-regulation occurred via fxr dependent suppression of the ntcp RXR:RAR response element. Moreover, cotransfected shp directly inhibited retinoid activation of the ntcp promoter. CONCLUSIONS: These studies show negative feedback regulation of ntcp by bile acid activated fxr via induction of shp. This novel regulatory pathway provides a means for coordinated down-regulation of bile acid import and synthesis, thereby protecting the hepatocyte from bile acid-mediated damage in cholestatic conditions. PMID- 11438504 TI - Angiotensin-converting enzyme inhibition attenuates the progression of rat hepatic fibrosis. AB - BACKGROUND AND AIMS: There is a significant relationship between inheritance of high transforming growth factor (TGF)-beta1 and angiotensinogen-producing genotypes and the development of progressive hepatic fibrosis in patients with chronic hepatitis C. In cardiac and renal fibrosis, TGF-beta1 production may be enhanced by angiotensin II, the principal effector molecule of the renin angiotensin system. The aim of the present study was to determine the effects of the angiotensin-converting enzyme inhibitor, captopril, on the progression of hepatic fibrosis in the rat bile duct ligation model. METHODS: Rats were treated with captopril (100 mg. kg(-1). day(-1)) commencing 1 or 2 weeks after bile duct ligation. Animals with bile duct ligation only and sham-operated animals served as controls. Four weeks after bile duct ligation, indices of fibrosis were assessed. RESULTS: Captopril treatment significantly reduced hepatic hydroxyproline levels, mean fibrosis score, steady state messenger RNA levels of TGF-beta1 and procollagen alpha1(I), and matrix metalloproteinase 2 and 9 activity. CONCLUSIONS: Captopril significantly attenuates the progression of hepatic fibrosis in the rat bile duct ligation model, and its effectiveness should be studied in human chronic liver diseases associated with progressive fibrosis. PMID- 11438505 TI - Proinflammatory cytokines inhibit secretion in rat bile duct epithelium. AB - BACKGROUND AND AIMS: Cholestatic disorders often are associated with portal inflammation, but whether or how inflammation contributes to cholestasis is unknown. Thus we studied the effects of proinflammatory cytokines on bile duct epithelia secretory mechanisms. METHODS: Isolated bile duct units (IBDUs) were cultured with interleukin (IL)-6, interferon gamma, tumor necrosis factor (TNF) alpha, and IL-1 alone or in combination. Ductular secretion was measured using video-optical planimetry. Bicarbonate and Cl(-) transport were assessed microfluorimetric measuring pH(i) (BCECF) and [Cl(-)](i) transients (MEQ). Expression of Cl(-)/HCO(3)(-) exchanger (AE-2), cystic fibrosis transmembrane conductance regulator (CFTR), and the secretin receptor (SR) were assessed by ribonuclease protection assay. Cellular cyclic adenosine monophosphate (cAMP) levels were studied by enzymatic immunoassay. Paracellular permeability was assessed using fluorescein-labeled dextrans (FD) in cholangiocyte monolayers (NRC 1). RESULTS: Although not effective when given alone, each combination of IL-6, interferon gamma, IL-1, and TNF-alpha inhibited secretion in IBDU. Cytokines inhibited cAMP formation, AE-2 activity, and cyclic AMP-dependent Cl(-) efflux, but not that induced by purinergic agonists. AE-2 gene expression was unaffected by proinflammatory cytokines, whereas CFTR and SR expression was increased. In addition, paracellular transit of FD across NRC-1 monolayers was increased. CONCLUSIONS: Inflammatory cytokines inhibit cAMP-dependent fluid secretion in cholangiocytes and impair the barrier functions of biliary epithelia. These changes may represent the molecular mechanisms by which inflammation leads to ductular cholestasis in vivo. PMID- 11438506 TI - Effects of ursodeoxycholic and cholic acid feeding on hepatocellular transporter expression in mouse liver. AB - BACKGROUND AND AIMS: Cholestasis is associated with retention of potentially toxic bile acids and alterations in hepatocellular transporter expression. Conversely, nontoxic ursodeoxycholic acid (UDCA) stimulates bile secretion and counteracts cholestasis. This study aimed to determine the effects of UDCA and cholic acid (CA) on the expression of hepatocellular transporters for bile acids (Ntcp, Bsep), organic anions (Oatp1, Mrp2), organic cations (Mdr1a/b), and phospholipids (Mdr2) in mouse liver. METHODS: Bile flow/composition was analyzed in UDCA- or CA-fed mice. Transporter expression was studied by reverse transcription polymerase chain reaction, Western blotting, and immunofluorescence microscopy. RESULTS: UDCA had no effect on basolateral Ntcp and down-regulated Oatp1, whereas canalicular Bsep and Mrp2 were up-regulated. CA down-regulated basolateral Ntcp and Oatp1, whereas canalicular Bsep, Mrp2, and Mdr1a/b were up regulated. Neither UDCA nor CA affected Mdr2 expression. Both UDCA and CA stimulated biliary bile acid and glutathione excretion, although only CA increased phospholipid and cholesterol excretion. CONCLUSIONS: Down-regulation of basolateral and up-regulation of canalicular transporters in response to CA may represent a defense mechanism, in an attempt to prevent hepatocellular accumulation of potentially toxic bile acids. The therapeutic effects of UDCA may be caused in part by stimulation of canalicular transporter expression in the absence of hepatocellular toxicity. PMID- 11438507 TI - Cd40 ligand-deficient mice are protected against cerulein-induced acute pancreatitis and pancreatitis-associated lung injury. AB - BACKGROUND AND AIMS: The interactions between inflammatory cells and their mediators play important roles in many inflammatory processes, but their importance during acute experimental pancreatitis and pancreatitis-associated lung injury is unclear. To address the role of the interaction between CD40 and its ligand CD40L, molecules that mediate major immunoregulatory functions, pancreatitis was induced by administering supramaximal doses of cerulein in mice that do not express CD40L. METHODS: The severity of pancreatitis was measured by serum amylase activity, pancreatic edema, acinar cell necrosis, and pancreas myeloperoxidase activity (an indicator of neutrophil infiltration). Lung injury was quantitated by evaluating lung microvascular permeability and lung myeloperoxidase activity. RESULTS: In pancreatic tissue from control mice and cerulein-treated mice, the expression of both CD40 and CD40L was detected. Immunohistochemical analysis performed in isolated acini from wild-type pancreata showed that both CD40 and CD40L were expressed on the acinar cell surface. Interestingly, pancreatitis and pancreatitis-associated lung injury were markedly decreased in mice deficient in CD40L compared with wild-types. CONCLUSIONS: These observations indicate that CD40L plays an important proinflammatory role in pancreatitis and pancreatitis-associated lung injury. PMID- 11438508 TI - American Gastroenterological Association medical position statement: hereditary colorectal cancer and genetic testing. AB - This document presents the official recommendations of the American Gastroenterological Association (AGA) on Hereditary Colorectal Cancer and Genetic Testing. It was approved by the Clinical Practice and Practice Economics Committee on March 20, 2001, and the AGA Governing Board on April 18, 2001. PMID- 11438509 TI - AGA technical review on hereditary colorectal cancer and genetic testing. AB - This literature review and the recommendations therein were prepared for the American Gastroenterological Association (AGA) Clinical Practice and Practice Economics Committee. The paper was approved by the Committee on March 20, 2001, and by the AGA Governing Board on April 18, 2001. PMID- 11438510 TI - Esophageal manometry prior to antireflux surgery: required, preferred, or even needed? PMID- 11438511 TI - Good cholesterol, bad cholesterol: role of oxysterols in biliary tract diseases. PMID- 11438512 TI - Bile-ology 101. PMID- 11438513 TI - Virtual virtual colonoscopy? PMID- 11438514 TI - Lamivudine for hepatitis B: the end of the beginning? PMID- 11438515 TI - Mutant salmonella as vectors for gene therapy. PMID- 11438517 TI - Functional differences of tau isoforms containing 3 or 4 C-terminal repeat regions and the influence of oxidative stress. AB - We report functional differences between tau isoforms with 3 or 4 C-terminal repeats and a difference in susceptibility to oxidative conditions, with respect to the regulation of microtubule dynamics in vitro and tau-microtubule binding in cultured cells. In the presence of dithiothreitol in vitro, a 3-repeat tau isoform promotes microtubule nucleation, reduces the tubulin critical concentration for microtubule assembly, and suppresses dynamic instability. Under non-reducing conditions, threshold concentrations of 3-repeat tau and tubulin exist below which this isoform still promotes microtubule nucleation and assembly but fails to reduce the tubulin critical concentration or suppress dynamic instability; above these threshold concentrations, amorphous aggregates of 3 repeat tau and tubulin can be produced at the expense of microtubule formation. A 4-repeat tau isoform is less sensitive to the oxidative potential of the environment, behaving under oxidative conditions similarly to the 3-repeat isoform under reducing conditions. Under conditions of oxidative stress, in Chinese hamster ovary cells stably expressing either 3- or 4-repeat tau, 3-repeat tau disassociates from microtubules more readily than the 4-repeat isoform, and tau-containing high molecular weight aggregates are preferentially observed in lysates from the Chinese hamster ovary cells expressing 3-repeat tau, indicating greater susceptibility of 3-repeat tau to oxidative conditions, compared with 4 repeat tau in vivo. PMID- 11438518 TI - Direct interaction of the Rab3 effector RIM with Ca2+ channels, SNAP-25, and synaptotagmin. AB - To define the role of the Rab3-interacting molecule RIM in exocytosis we searched for additional binding partners of the protein. We found that the two C(2) domains of RIM display properties analogous to those of the C(2)B domain of synaptotagmin-I. Thus, RIM-C(2)A and RIM-C(2)B bind in a Ca(2+)-independent manner to alpha1B, the pore-forming subunit of N-type Ca(2+) channels (EC(50) = approximately 20 nm). They also weakly interact with the alpha1C but not the alpha1D subunit of L-type Ca(2+) channels. In addition, the C(2) domains of RIM associate with SNAP-25 and synaptotagmin-I. The binding affinities for these two proteins are 203 and 24 nm, respectively, for RIM-C(2)A and 224 and 16 nm for RIM C(2)B. The interactions of the C(2) domains of RIM with SNAP-25 and synaptotagmin I are modulated by Ca(2+). Thus, in the presence of Ca(2+) (EC(50) = approximately 75 microm) the interaction with synaptotagmin-I is increased, whereas SNAP-25 binding is reduced. Synaptotagmin-I binding is abolished by mutations in two positively charged amino acids in the C(2) domains of RIM and by the addition of inositol polyphosphates. We propose that the Rab3 effector RIM is a scaffold protein that participates through its multiple binding partners in the docking and fusion of secretory vesicles at the release sites. PMID- 11438519 TI - HIV Nef-mediated cellular phenotypes are differentially expressed as a function of intracellular Nef concentrations. AB - Nef is a regulatory protein encoded by the genome of both human and simian immunodeficiency virus. Its expression in T cells leads to CD4 and major histocompatibility complex class I modulation and either enhancement or suppression of T cell activation. How this viral protein achieves multiple and at times opposing activities has been unclear. Through direct measurements of Nef and the Nef-GFP fusion protein, we find that these events are mediated by different Nef concentrations. Relative to the intracellular concentration that down-modulates surface CD4, an order of magnitude increase in Nef-GFP expression is required for a comparable modulation of major histocompatibility complex class I, and a further 3-fold increase is necessary to suppress T cell activation. PMID- 11438520 TI - Mutations of either or both Cys876 and Cys888 residues of sarcoplasmic reticulum Ca2+-ATPase result in a complete loss of Ca2+ transport activity without a loss of Ca2+-dependent ATPase activity. Role of the CYS876-CYS888 disulfide bond. AB - Disulfide-containing peptides in pepsin digest of sarcoplasmic reticulum vesicles were identified by using a fluorogenic thiol-specific reagent 4-fluoro-7 sulfamoylbenzofurazan and a reductant tributylphosphine. Sequencing of the purified peptides revealed the presence of a Cys(876)-Cys(888) disulfide bond on the luminal loop connecting the 7th and 8th transmembrane helices (loop 7-8) of the Ca(2+)-ATPase (SERCA1a). We substituted either or both of these cysteine residues with alanine and made three mutants (C876A, C888A, C876A/C888A), in which the disulfide bond is disrupted. The mutants and the wild type were expressed in COS-1 cells, and functional analysis was performed with the microsomes isolated from the cells. Electrophoresis performed under reducing and non-reducing conditions confirmed the presence of Cys(876)-Cys(888) disulfide bond in the expressed wild type. All the three mutants possessed high Ca(2+) ATPase activity. In contrast, no Ca(2+) transport activity was detected with these mutants. These mutants formed almost the same amount of phosphoenzyme intermediate as the wild type from ATP and from P(i). Detailed kinetic analysis showed that the three mutants hydrolyze ATP in the mechanism well accepted for the Ca(2+)-ATPase; activation of the catalytic site upon high affinity Ca(2+) binding, formation of ADP-sensitive phosphoenzyme, subsequent rate-limiting transition to ADP-insensitive phosphoenzyme, and hydrolysis of the latter phosphoenzyme. It is likely that the pathway for delivery of Ca(2+) from the binding sites into the lumen of vesicles is disrupted by disruption of the Cys(876)-Cys(888) disulfide bond, and therefore that the loop 7-8 having the disulfide bond is important for formation of the proper structure of the Ca(2+) pathway. PMID- 11438521 TI - Induction of neurite outgrowth in PC12 cells by alpha -phenyl-N-tert-butylnitron through activation of protein kinase C and the Ras-extracellular signal-regulated kinase pathway. AB - The spin trap alpha-phenyl-N-tert-butylnitron (PBN) is widely used for studies of the biological effects of free radicals. We previously reported the protective effects of PBN against ischemia-reperfusion injury in gerbil hippocampus by its activation of extracellular signal-regulated kinase (ERK) and suppression of both stress-activated protein kinase and p38 mitogen-activated protein kinase. In the present study, we found that PBN induced neurite outgrowth accompanied by ERK activation in PC12 cells in a dose-dependent manner. The induction of neurite outgrowth was inhibited significantly not only by transient transfection of PC12 cells with dominant negative Ras, but also by treatment with mitogen-activated protein kinase/ERK kinase inhibitor PD98059. The activation of receptor tyrosine kinase TrkA was not involved in PBN-induced neurite outgrowth. A protein kinase C (PKC) inhibitor, GF109203X, was found to inhibit neurite outgrowth. The activation of PKCepsilon was observed after PBN stimulation. PBN-induced neurite outgrowth and ERK activation were counteracted by the thiol-based antioxidant N acetylcysteine. From these results, it was concluded that PBN induced neurite outgrowth in PC12 cells through activation of the Ras-ERK pathway and PKC. PMID- 11438522 TI - Actin cytoskeletal association of cytohesin-1 is regulated by specific phosphorylation of its carboxyl-terminal polybasic domain. AB - Cell adhesion mediated by integrin receptors is controlled by intracellular signal transduction cascades. Cytohesin-1 is an integrin-binding protein and guanine nucleotide exchange factor that activates binding of the leukocyte integrin leukocyte function antigen-1 to its ligand, intercellular adhesion molecule 1. Cytohesin-1 bears a carboxyl-terminal pleckstrin homology domain that aids in reversible membrane recruitment and functional regulation of the protein. Although phosphoinositide-dependent membrane attachment of cytohesin-1 is mediated primarily by the pleckstrin homology domain, this function is further strengthened by a short carboxyl-terminal polybasic amino acid sequence. We show here that a serine/threonine motif within the short polybasic stretch of cytohesin-1 is phosphorylated by purified protein kinase C delta in vitro. Furthermore, the respective residues are also found to be phosphorylated after phorbol ester stimulation in vivo. Biochemical and functional analyses show that phosphorylated cytohesin-1 is able to tightly associate with the actin cytoskeleton, and we further demonstrate that phosphorylation of the protein is required for maximal leukocyte function antigen-1-mediated adhesion of Jurkat cells to intercellular adhesion molecule 1. These data suggest that both phosphatidylinositol 3-kinase and protein kinase C-dependent intracellular pathways that stimulate beta(2)-integrin-mediated adhesion of T lymphocytes converge on cytohesin-1 as functional integrator. PMID- 11438523 TI - Peroxisome proliferator-activated receptor-gamma-independent inhibition of macrophage activation by the non-thiazolidinedione agonist L-796,449. Comparison with the effects of 15-deoxy-delta(12,14)-prostaglandin J(2). AB - The effects of L-796,449 (3-chloro-4-(3-(3-phenyl-7-propylbenzofuran-6 yloxy)propylthio)phenylacetic acid; referred to henceforth as compound G), a thiazolidinedione-unrelated peroxisome proliferator activated-receptor-gamma (PPAR-gamma) agonist, on early signaling in lipopolysaccharide-activated RAW 264.7 macrophages were analyzed and compared with those elicited by 15-deoxy Delta(12,14)-prostaglandin J(2) and the thiazolidinedione rosiglitazone. Compound G inhibited the activation of nuclear factor kappa B through the impairment of the targeting and degradation of I kappa B proteins and promoted a redistribution of I kappa B alpha and I kappa B beta in the nucleus of activated cells. Compound G inhibited I kappa B kinase (IKK) activity both in vivo and in vitro, suggesting a direct mechanism of interaction between this molecule and the IKK complex. The effect of compound G on IKK activity was independent of PPAR-gamma engagement because RAW 264.7 cells expressed negligible levels of this nuclear receptor, and rosiglitazone failed to mimic these actions. Moreover, treatment of activated macrophages with compound G enhanced the synthesis of superoxide anion, which, in combination with the NO produced under activation conditions, triggered apoptosis through the intracellular synthesis of peroxynitrite. These results suggest that compound G might contribute to the resolution of inflammation by favoring the induction of apoptosis through mechanisms independent of PPAR-gamma engagement. PMID- 11438524 TI - The human CD8 coreceptor effects cytotoxic T cell activation and antigen sensitivity primarily by mediating complete phosphorylation of the T cell receptor zeta chain. AB - Recognition of antigen by cytotoxic T lymphocytes (CTL) is determined by interaction of both the T cell receptor and its CD8 coreceptor with peptide-major histocompatibility complex (pMHC) class I molecules. We examine the relative roles of these receptors in the activation of human CTL using mutations in MHC class I designed to diminish or abrogate the CD8/pMHC interaction. We use surface plasmon resonance to determine that point mutation of the alpha3 loop of HLA A2 abrogates the CD8/pMHC interaction without affecting the affinity of the T cell receptor/pMHC interaction. Antigen-presenting cells expressing HLA A2 which does not bind to CD8 fail to activate CTL at any peptide concentration. Comparison of CTL activation by targets expressing HLA A2 with normal, abrogated, or diminished CD8/pMHC interaction show that the CD8/pMHC interaction enhances sensitivity to antigen. We determine that the biochemical basis for coreceptor dependence is the activation of the 23-kDa phosphoform of the CD3zeta chain. In addition, we produce mutant MHC class I multimers that specifically stain but do not activate CTL. These reagents may prove useful in circumventing undesirable activation related perturbation of intracellular processes when pMHC multimers are used to phenotype antigen-specific CD8+ lymphocytes. PMID- 11438525 TI - Silencer activity of NFATc2 in the interleukin-12 receptor beta 2 proximal promoter in human T helper cells. AB - Interleukin 12 (IL-12) is a potent enhancer of interferon gamma production by activated T cells. The high-affinity IL-12 receptor (IL-12R) is a heterodimer of a beta1 and a beta2 subunit. Expression of the signaling IL-12Rbeta2 chain is usually low, as compared with the more abundant beta1 chain, and may be rate limiting for IL-12 sensitivity. Little is known about the mechanisms controlling IL-12Rbeta2 gene expression. Reporter gene assays in IL-12Rbeta2-expressing Jurkat cells showed that truncation of the region from -151 to -61 abrogated promoter activity. The proximal promoter region does not contain a typical TATA box, suggesting a role for SP-1. Indeed, mutagenesis of the -63 SP-1 consensus site decreased transcription by 50%. Electrophoretic mobility shift experiments confirmed the binding of SP-1 and SP-3 at this site. In contrast, truncation of 252 to -192 increased promoter activity. Likewise, mutagenesis of the consensus nuclear factor of activated T cells site at -206 increased promoter activity by 70%, suggesting silencer activity of this element. Electrophoretic mobility shift experiments with primary Th (T helper) cells showed the formation of a specific, T-cell receptor-inducible complex at this site that is sensitive to cyclosporin A and supershifted with anti-NFATc2 in both Th1 and Th2 cells. Accordingly, cyclosporin A dose-dependently increased IL-12Rbeta2 mRNA expression. These first data on IL-12Rbeta2 gene regulation indicate a TATA-less promoter, depending on SP-1/SP-3 transcription factors, and a negative regulatory NFAT element at -206. This element may contribute to the overall low level of IL-12Rbeta2 expression on Th cells. PMID- 11438526 TI - Conformational dynamics underlie the activity of the auxin-binding protein, Nt abp1. AB - The auxin-binding protein 1 (ABP1) has been proposed to be involved in the perception of the phytohormone at the plasma membrane. Site-directed mutagenesis was performed on highly conserved residues at the C terminus of ABP1 to investigate their relative importance in protein folding and activation of a functional response at the plasma membrane. Detailed analysis of the dynamic interaction of the wild-type ABP1 and mutated proteins with three distinct monoclonal antibodies recognizing conformation-dependent epitopes was performed by surface plasmon resonance. The influence of auxin on these interactions was also investigated. The Cys(177) as well as Asp(175) and Glu(176) were identified as critical residues for ABP1 folding and action at the plasma membrane. On the contrary, the C-terminal KDEL sequence was demonstrated not to be essential for auxin binding, interaction with the plasma membrane, or activation of the transduction cascade although it does appear to be involved in the stability of ABP1. Taken together, the results confirmed that ABP1 conformational change is the critical step for initiating the signal from the plasma membrane. PMID- 11438527 TI - Crystal structure of human epidermal growth factor and its dimerization. AB - Epidermal growth factor (EGF) is a typical growth-stimulating peptide and functions by binding to specific cell-surface receptors and inducing dimerization of the receptors. Little is known about the molecular mechanism of EGF-induced dimerization of EGF receptors. The crystal structure of human EGF has been determined at pH 8.1. There are two human EGF molecules A and B in the asymmetric unit of the crystals, which form a potential dimer. Importantly, a number of residues known to be indispensable for EGF binding to its receptor are involved in the interface between the two EGF molecules, suggesting a crucial role of EGF dimerization in the EGF-induced dimerization of receptors. In addition, the crystal structure of EGF shares the main features of the NMR structure of mouse EGF determined at pH 2.0, but structural comparisons between different models have revealed new detailed features and properties of the EGF structure. PMID- 11438528 TI - Binding of p300/CBP co-activators by polyoma large T antigen. AB - Small DNA tumor viruses such as simian virus 40 (SV40) and polyomavirus (Py) take advantage of host cell proteins to transcribe and replicate their DNA. Interactions between the viral T antigens and host proteins result in cell transformation and tumor induction. Large T antigen of SV40 interacts with p53, pRb/p107/p130 family members, and the cyclic AMP-responsive element-binding protein (CREB)-binding protein (CBP)/p300. Py large T antigen is known to interact only with pRb and p300 among these proteins. Here we report that Py large T binds to CBP in vivo and in vitro. In co-transfection assays, Py large T inhibits the co-activation functions of CBP/p300 in CREB-mediated transactivation but not in NF-kappa B-mediated transactivation. p53 appears not to be involved in the functions of CREB-mediated transactivation and is not essential for large T:CBP interaction. Mutations introduced into a region of Py large T with homology to adenovirus E1A and SV40 large T prevent binding to the co-activators. These mutant large T antigens fail to inhibit CREB-mediated transactivation. The CBP/p300-binding Py mutants are able to transform established rat embryo fibroblasts but are restricted in their ability to induce tumors in the newborn mouse, indicating that interaction of large T with the co-activators may be essential for virus replication and spread in the intact host. PMID- 11438529 TI - Definition of the extended substrate specificity determinants for beta-tryptases I and II. AB - Tryptases betaI and betaII were heterologously expressed and purified in yeast to functionally characterize the substrate specificity of each enzyme. Three positional scanning combinatorial tetrapeptide substrate libraries were used to determine the primary and extended substrate specificity of the proteases. Both enzymes have a strict primary preference for cleavage after the basic amino acids, lysine and arginine, with only a slight preference for lysine over arginine. betaI and betaII tryptase share similar extended substrate specificity, with preference for proline at P4, preference for arginine or lysine at P3, and P2 showing a slight preference for asparagine. Measurement of kinetic constants with multiple substrates designed for beta-tryptases reveal that selectivity is highly dependent on ground state substrate binding. Coupled with the functional determinants, structural determinants of tryptase substrate specificity were identified. Molecular docking of the preferred substrate sequence to the three dimensional tetrameric tryptase structure reveals a novel extended substrate binding mode that involves interactions from two adjacent protomers, including P4 Thr-96', P3 Asp-60B' and Glu-217, and P1 Asp-189. Based on the determined substrate information, a mechanism-based tetrapeptide-chloromethylketone inhibitor was designed and shown to be a potent tryptase inhibitor. Finally, the cleavage sites of several physiologically relevant substrates of beta-tryptases show consistency with the specificity data presented here. PMID- 11438530 TI - Altering the DNA-binding specificity of the yeast Matalpha 2 homeodomain protein. AB - Homeodomain proteins are a highly conserved class of DNA-binding proteins that are found in virtually every eukaryotic organism. The conserved mechanism that these proteins use to bind DNA suggests that there may be at least a partial DNA recognition code for this class of proteins. To test this idea, we have investigated the sequence-specific requirements for DNA binding and repression by the yeast alpha2 homeodomain protein in association with its cofactors, Mcm1 and Mata1. We have determined the contribution for each residue in the alpha2 homeodomain that contacts the DNA in the co-crystal structures of the protein. We have also engineered mutants in the alpha2 homeodomain to alter the DNA-binding specificity of the protein. Although we were unable to change the specificity of alpha2 by making substitutions at residues 47, 54, and 55, we were able to alter the DNA-binding specificity by making substitutions at residue 50 in the homeodomain. Since other homeodomain proteins show similar changes in specificity with substitutions at residue 50, this suggests that there is at least a partial DNA recognition code at this position. PMID- 11438531 TI - Nrl and Sp nuclear proteins mediate transcription of rod-specific cGMP phosphodiesterase beta-subunit gene: involvement of multiple response elements. AB - cGMP-phosphodiesterase (PDE) is the key effector in rod photoreceptor signal transduction. Mutations in the gene encoding its catalytic beta-subunit (beta PDE) cause retinal degenerations leading to blindness. We report that the short 93 to +53 sequence in the upstream region of this gene is sufficient for beta-PDE transcription in both Y79 human retinoblastoma cells and Xenopus embryo heads maintained ex vivo. This sequence also functions as a minimal rod-specific promoter in transgenic Xenopus tadpoles. The Nrl transcription factor binds in vitro to the betaAp1/NRE regulatory element located within this region and transactivates it when overexpressed in nonretinal 293 embryonic kidney cells. We also found a G/C-rich activator element, beta/GC, important for promoter activity in Y79 retinoblastoma cells and Xenopus embryos. Both the ubiquitous Sp1 and the central nervous system-specific Sp4 transcription factors are expressed in retina and interact with this element in vitro. Electrophoretic mobilities of beta/GC Y79 nuclear protein complexes are altered by antibodies against Sp1 and Sp4. Thus, our results implicate Nrl, Sp1, and Sp4 in transcriptional regulation of the rod-specific minimal beta-PDE promoter. We also conclude that Xenopus laevis is an efficient system for analyzing the human beta-PDE promoter and may be used to study other human retinal genes ex vivo and in vivo. PMID- 11438532 TI - Two dimerization domains in the trans-activation response RNA-binding protein (TRBP) individually reverse the protein kinase R inhibition of HIV-1 long terminal repeat expression. AB - Trans-activation response (TAR) RNA-binding protein (TRBP) is a cellular protein that binds to the human immunodeficiency virus-1 (HIV-1) TAR element RNA. It has two double-stranded RNA binding domains (dsRBDs), but only one is functional for TAR binding. TRBP interacts with the interferon-induced protein kinase R (PKR) and inhibits its activity. We used the yeast two-hybrid assay to map the interaction sites between the two proteins. We show that TRBP and PKR-N (178 first amino acids of PKR) interact with PKR wild type and inhibit the PKR-induced yeast growth defect in this assay. We characterized two independent PKR-binding sites in TRBP. These sites are located in each dsRBD in TRBP, indicating that PKR TRBP interaction does not require the RNA binding activity present only in dsRBD2. TRBP and its fragments that interact with PKR reverse the PKR-induced suppression of HIV-1 long terminal repeat expression. In addition, TRBP activates the HIV-1 long terminal repeat expression to a larger extent than the addition of each domain. These data suggest that TRBP activates gene expression in PKR dependent and PKR-independent manners. PMID- 11438533 TI - Cellular membrane composition defines A beta-lipid interactions. AB - Alzheimer's disease pathology has demonstrated amyloid plaque formation associated with plasma membranes and the presence of intracellular amyloid-beta (A beta) accumulation in specific vesicular compartments. This suggests that lipid composition in different compartments may play a role in A beta aggregation. To test this hypothesis, we have isolated cellular membranes from human brain to evaluate A beta 40/42-lipid interactions. Plasma, endosomal, lysosomal, and Golgi membranes were isolated using sucrose gradients. Electron microscopy demonstrated that A beta fibrillogenesis is accelerated in the presence of plasma and endosomal and lysosomal membranes with plasma membranes inducing an enhanced surface organization. Alternatively, interaction of A beta with Golgi membranes fails to progress to fibril formation, suggesting that A beta-Golgi head group interaction stabilizes A beta. Fluorescence spectroscopy using the environment-sensitive probes 1,6-diphenyl-1,3,5-hexatriene, laurdan, N epsilon-dansyl-L-lysine, and merocyanine 540 demonstrated variations in the inherent lipid properties at the level of the fatty acyl chains, glycerol backbone, and head groups, respectively. Addition of A beta 40/42 to the plasma and endosomal and lysosomal membranes decreases the fluidity not only of the fatty acyl chains but also the head group space, consistent with A beta insertion into the bilayer. In contrast, the Golgi bilayer fluidity is increased by A beta 40/42 binding which appears to result from lipid head group interactions and the production of interfacial packing defects. PMID- 11438534 TI - Chloroplast glyceraldehyde-3-phosphate dehydrogenase contains a single disulfide bond located in the C-terminal extension to the B subunit. AB - Mass mapping analysis based on cyanylation and CN-induced cleavage indicates that the two cysteine residues in the C-terminal extension of the B subunit of the light-activated pea leaf chloroplast glyceraldehyde-3-phosphate dehydrogenase form a disulfide bond. No evidence was found for a disulfide bond in the A subunit, nor was there any indication of a second disulfide bond in the B subunit. The availability of the structure of the extended glyceraldehyde-3 phosphate dehydrogenase from the archaeon Sulfolobus solfataricus allows modeling of the B subunit. As modeled, the two cysteine residues in the extension are positioned to form an interdomain disulfide cross-link. PMID- 11438535 TI - The apoptotic regulatory protein ARC (apoptosis repressor with caspase recruitment domain) prevents oxidant stress-mediated cell death by preserving mitochondrial function. AB - ARC is an apoptotic regulatory protein expressed almost exclusively in myogenic cells. It contains a caspase recruitment domain (CARD) through which it has been shown to block the activation of some initiator caspases. Because ARC also blocks caspase-independent events associated with apoptosis, such as hypoxia-induced cytochrome c release, we examined its role in cell death triggered by exposure to hydrogen peroxide (H(2)O(2)) in the myogenic cell line, H9c2. Cell death in this model was caspase-independent and characterized by dose-dependent reduction in ARC expression accompanied by disruption of the mitochondrial membrane potential (Delta psi(m)) and loss of plasma membrane integrity, typical of necrotic cell death. Ectopic expression of ARC prevented both H(2)O(2)-induced mitochondrial dysfunction and cell death without affecting the stress kinase response, suggesting that ARCs protective effects were downstream of early signaling events and not due to quenching of H(2)O(2). ARC was also effective in blocking H(2)O(2) induced loss of membrane integrity and/or disruption of Delta psi(m) in two human cell lines in which it is not normally expressed. These results demonstrate that, in addition to its ability to block caspase-dependent and -independent events in apoptosis, ARC also prevents necrosis-like cell death via the preservation of mitochondrial function. PMID- 11438536 TI - Characterization of two evolutionarily conserved, alternatively spliced nuclear phosphoproteins, NFAR-1 and -2, that function in mRNA processing and interact with the double-stranded RNA-dependent protein kinase, PKR. AB - We report here the isolation and characterization of two proteins, NFAR-1 and -2, which were isolated through their ability to interact with the dsRNA-dependent protein kinase, PKR. The NFAR proteins, of 90 and 110 kDa, are derived from a single gene through alternative splicing and are evolutionarily conserved nuclear phosphoproteins that interact with double-stranded RNA. Both NFAR-1 and -2 are phosphorylated by PKR, reciprocally co-immunoprecipitate with PKR, and colocalize with the kinase in a diffuse nuclear pattern within the cell. Transfection studies indicate that the NFARs regulate gene expression at the level of transcription, probably during the processing of pre-mRNAs, an activity that was increased in fibroblasts lacking PKR. Subsequent functional analyses indicated that amino acids important for NFAR's activity were localized to the C terminus of the protein, a region that was found to specifically interact with FUS and SMN, proteins also known as regulators of RNA processing. Accordingly, both NFARs were found to associate with both pre-mRNAs and spliced mRNAs in post transcriptional studies, similar to the known splicing factor ASF/SF-2. Collectively, our data indicate that the NFARs may facilitate double-stranded RNA regulated gene expression at the level of post-transcription and possibly contribute to host defense-related mechanisms in the cell. PMID- 11438537 TI - The first transmembrane domain of the P2X receptor subunit participates in the agonist-induced gating of the channel. AB - Based on pharmacological properties, the P2X receptor family can be subdivided into those homo-oligomers that are sensitive to the ATP analog alphabeta methylene ATP(alphabetameATP) (P2X(1) and P2X(3)) and those that are not (P2X(2), P2X(4), P2X(5), P2X(6), and P2X(7)). We exploited this dichotomy through the construction of chimeric receptors and site-directed mutagenesis in order to identify domains responsible for these differences in the abilities of extracellular agonists to gate P2X receptors. Replacement of the extracellular domain of the alphabetameATP-sensitive rat P2X(1) subunit with that of the alphabetameATP-insensitive rat P2X(2) subunit resulted in a receptor that was still alphabetameATP-sensitive, suggesting a non-extracellular domain was responsible for the differential gating of P2X receptors by various agonists. Replacement of the first transmembrane domain of the rat P2X(2) subunit with one from an alphabetameATP-sensitive subunit (either rat P2X(1) or P2X(3) subunit) converted the resulting chimera to alphabetameATP sensitivity. This conversion did not occur when the first transmembrane domain came from a non-alphabetameATP sensitive subunit. Site-directed mutagenesis indicated that the C-terminal portion of the first transmembrane domain was important in determining the agonist selectivity of channel gating for these chimeras. These results suggest that the first transmembrane domain plays an important role in the agonist operation of the P2X receptor. PMID- 11438538 TI - Differential regulation of the aldehyde dehydrogenase 1 gene in embryonic chick retina and liver. AB - Aldehyde dehydrogenase (ALDH1) is highly expressed in the dorsal cells of the undifferentiated retina, where it has been proposed to play a role in the formation of a retinoic acid gradient along the ventrodorsal axis. In contrast to the retina, ALDH1 levels increase with differentiation in the liver and remain elevated in the adult tissue. To understand the molecular basis for differential expression of ALDH1 during development, we characterized the ALDH1 transcripts expressed in chick retina and liver. By sequencing, primer extension, and S1 nuclease analysis, we show that retina ALDH1 mRNA has an additional 300 nucleotides of 5'-untranslated sequence resulting from the transcription of two 5' noncoding exons. There is a 24-29-kilobase pair (kb) gap between exons 1 and 2 and a 290-base pair gap between exons 2 and 3. Exon 3, which contains the ALDH1 start codon, represents the first exon of the liver transcript. Using a reporter gene assay, we have identified tissue-specific regulatory elements that govern ALDH1 expression in primary retina and liver cultures. Constructs with >1.6 kb of DNA flanking the 5'-end of exon 1 showed elevated activity in retinal cultures but only basal activity in liver cultures. In contrast, constructs with <1 kb of 5'-flanking DNA were active in both retina and liver cultures. Our results suggest that an important mechanism for the control of ALDH1 transcriptional activity is through the presence of inhibitory elements located 0.7-1.6 kb upstream of the ALDH1 gene. DNase I footprint analysis reveal four sites of protein-DNA interaction within this region, one of which is specific to the liver and corresponds to a NF-kappaB/Rel binding site. PMID- 11438539 TI - Cloning and characterization of three differentially expressed peroxidoxin genes from Leishmania chagasi. Evidence for an enzymatic detoxification of hydroxyl radicals. AB - Antioxidants have been implicated in protecting cells from oxygen radicals produced as a result of aerobic metabolism and in response to foreign pathogens by phagocytic cells. The mechanisms allowing pathogens to withstand the toxic prooxidant environment within the phagolysosome are poorly understood. We have cloned and characterized three antioxidant genes belonging to the 2-Cys family of peroxidoxins from Leishmania chagasi that may prove to provide these parasites with an enhanced defense mechanism against toxic oxidants. The 5'-untranslated regions and coding regions of each gene are highly conserved, whereas the 3' untranslated regions have diverged significantly. L. chagasi peroxidoxin 1 (LcPxn1) is predominantly expressed in the amastigote stage, whereas LcPxn2 and LcPxn3 are expressed mainly in the promastigote stage, with LcPxn3 being far less abundant than LcPxn2. LcPxn2 and LcPxn3 possess a nine-amino acid extension at the carboxyl terminus, which LcPxn1 lacks. LcPxn1 appears to exist as high molecular weight multimers in vivo, and recombinant LcPxn1 was shown to detoxify hydrogen peroxide and alkyl hydroperoxides. We also present strong evidence that recombinant LcPxn1 can enzymatically detoxify hydroxyl radicals, an activity never before clearly demonstrated for a protein. PMID- 11438540 TI - Nuclear factor 90 is a substrate and regulator of the eukaryotic initiation factor 2 kinase double-stranded RNA-activated protein kinase. AB - Nuclear factor 90 (NF90) is a member of an expanding family of double-stranded (ds) RNA-binding proteins thought to be involved in gene expression. Originally identified in complex with nuclear factor 45 (NF45) as a sequence-specific DNA binding protein, NF90 contains two double stranded RNA-binding motifs (dsRBMs) and interacts with highly structured RNAs as well as the dsRNA-activated protein kinase, PKR. In this report, we characterize the biochemical interactions between these two dsRBM containing proteins. NF90 binds to PKR through two independent mechanisms: an RNA-independent interaction occurs between the N terminus of NF90 and the C-terminal region of PKR, and an RNA-dependent interaction is mediated by the dsRBMs of the two proteins. Co-immunoprecipitation analysis demonstrates that NF90, NF45, and PKR form a complex in both nuclear and cytosolic extracts, and both proteins serve as substrates for PKR in vitro. NF90 is phosphorylated by PKR in its RNA-binding domain, and this reaction is partially blocked by the NF90 N terminal region. The C-terminal region also inhibits PKR function, probably through competitive binding to dsRNA. A model for NF90-PKR interactions is proposed. PMID- 11438541 TI - The di-aromatic pentapeptide repeats of the human peroxisome import receptor PEX5 are separate high affinity binding sites for the peroxisomal membrane protein PEX14. AB - PEX5 functions as a mobile import receptor for peroxisomal matrix proteins with a peroxisomal targeting signal 1 (PTS1). A critical step within the PTS1-import pathway is the interaction between PEX5 and the peroxisome membrane-associated protein PEX14. Based on two-hybrid analyses in mammalian cells and complementary in vitro binding assays, we demonstrate that the evolutionarily conserved pentapeptide repeat motifs, WX(E/D/Q/A/S)(E/D/Q)(F/Y), in PEX5 bind to PEX14 with high affinity. The results obtained indicate that each of the seven di-aromatic pentapeptides of human PEX5 interacts separately at the same binding site in the N terminus of PEX14 with equilibrium dissociation constants in the low nanomolar range. Mutational analysis of the PEX14-binding motifs reveals that the conserved aromatic amino acids at position 1 or 5 are essential for high affinity binding. We propose that the side chains of the aromatic amino acids are in close proximity as part of an amphipathic alpha-helix and together form hydrophobic anchors for binding PEX5 to individual PEX14 molecules. PMID- 11438542 TI - The DNA glycosylase T:G mismatch-specific thymine DNA glycosylase represses thyroid transcription factor-1-activated transcription. AB - The transcription factor thyroid transcription factor-1 (TTF-1) is a homeodomain containing protein that belongs to the NK2 family of genes involved in organogenesis. TTF-1 is required for normal development of the forebrain, lung, and thyroid. In a search for factors that regulate TTF-1 transcriptional activity, we isolated three genes (T:G mismatch-specific thymine DNA glycosylase (TDG), homeodomain-interacting protein kinase 2 (HIPK2), and Ajuba), whose products can interact with TTF-1 in yeast and in mammalian cells. TDG is an enzyme involved in base excision repair. In the present paper, we show that TDG acts as a strong repressor of TTF-1 transcriptional activity in a dose-dependent manner, while HIPK2 and Ajuba display no effect on TTF-1 activity, at least under the tested conditions. TDG-mediated inhibition occurs specifically on TTF-1 responsive promoters in thyroid and non thyroid cells. TDG associates with TTF-1 in mammalian cells through the TTF-1 carboxyl-terminal activation domain and is independent of the homeodomain. These findings reveal a previously unsuspected role for the repair enzyme TDG as a transcriptional repressor and open new routes toward the understanding of the regulation of TTF-1 transcriptional activity. PMID- 11438543 TI - Functional importance of Stat3 tetramerization in activation of the alpha 2 macroglobulin gene. AB - A tetrameric Stat3 complex was found to be essential in transfection experiments for maximal interleukin-6-inducible activation of alpha2-macroglobulin gene promoter. Stable tetramer formation of purified phosphorylated Stat3 was dependent on protein.protein interaction involving the N-terminal domain of Stat3. The functional importance of tetramer formation was shown by the decreased levels of transcriptional activation associated with hypomorphic mutations in N terminal residues. PMID- 11438544 TI - Roles of phosphatidylinositol 3-kinase in interferon-gamma-dependent phosphorylation of STAT1 on serine 727 and activation of gene expression. AB - STAT1 must be phosphorylated on serine 727 to be fully active in transcription. We show that phosphatidylinositol 3-kinase (PI3K) and its effector kinase Akt play an important role in the serine phosphorylation of STAT1 and in the activation of gene expression in response to interferon-gamma (IFN gamma). IFN gamma activates PI3K as well as Akt in a variety of cell lines. Specific inhibition of PI3K abrogates IFN gamma-induced, but not interleukin-1- or tumor necrosis factor-alpha-induced, phosphorylation of STAT1 on serine and reduces STAT1-dependent transcription and gene expression by approximately 7-fold. Constitutively active forms of PI3K or Akt activate and their dominant-negative derivatives inhibit STAT1-driven transactivation in response to IFN gamma. In addition to PI3K and Akt, JAK1, JAK2, and the tyrosine 440 STAT1 docking residue of IFNGR1 are required for STAT1 to be phosphorylated on serine. Taken together, these results suggest that the following events lead to the activation of STAT1 upon IFN gamma stimulation: 1) PI3K and Akt are activated by the occupied receptor and Tyr-440 is phosphorylated by the activated JAKs; 2) STAT1 docks to Tyr-440; and 3) Tyr-701 is phosphorylated by the JAKs and Ser-727 is phosphorylated by a kinase downstream of Akt. PMID- 11438545 TI - Ligand binding and hexacoordination in synechocystis hemoglobin. AB - A large and phylogenetically diverse group of organisms contain truncated hemoglobins, including the unicellular cyanobacterium Synechocystis (Pesce, A., Couture, M., Dewilde, S., Guertin, M., Yamauchi, K., Ascenzi, P., Moens, L., and Bolognesi, M. (2000) EMBO J. 19, 2424-2434). Synechocystis hemoglobin is also hexacoordinate, with a heme pocket histidine that reversibly coordinates the ligand binding site. Hexacoordinate hemoglobins are ubiquitous in plants and are now being identified in a diverse array of organisms including humans (Arredondo Peter, R., Hargrove, M. S., Moran, J. F., Sarath, G., and Klucas, R. V. (1998) Plant Physiol. 118, 1121-1125; Trent, J. T., III, Watts, R. A., and Hargrove, M. S. (2001) J. Biol. Chem. 276, 30106-30110). Rate constants for association and dissociation of the hexacoordinating amino acid side chain in Synechocystis hemoglobin have been measured along with bimolecular rate constants for association of oxygen and carbon monoxide following laser flash photolysis. These values were compared with ligand binding initiated by rapid mixing. Site-directed mutagenesis was used to determine the roles of several heme pocket amino acids in facilitating hexacoordination and stabilizing bound oxygen. It is demonstrated that Synechocystis hemoglobin contains a very reactive binding site and that ligand migration through the protein is rapid. Rate constants for hexacoordination by His(46) are also large and facilitated by other heme pocket amino acids including Gln(43). PMID- 11438546 TI - Biological properties of poly-L-lysine-DNA complexes generated by cooperative binding of the polycation. AB - We have evaluated the effect of NaCl concentration on the mode of binding of poly L-lysine to DNA and the resulting structural and functional features of the condensed DNA particles using DNA precipitation, DNase I resistance, electron microscopy, and receptor-mediated gene transfer assays. At a high concentration of NaCl and in the presence of excess DNA, poly-L-lysine interacted with DNA cooperatively, fully condensing some of the DNA and leaving the rest of the DNA unbound. At low NaCl concentrations, poly-L-lysine molecules interacted with DNA in a noncooperative fashion, i.e. they bind randomly to the whole population of DNA molecules. Cooperative binding of poly-L-lysine to DNA occurred over a narrow range of NaCl concentrations, and the specific salt concentration depended on the length of the poly-L-lysine. The ability of condensed DNA to withstand digestion by DNase I was correlated with the structural features of the condensed DNA as determined by electron microscopy. Using our condensation procedure, cooperative binding of poly-L-lysine to DNA is a necessary prerequisite for the preparation of condensed DNA having a spherical shape and a diameter of 15-30 nm. Condensed DNA, containing galactosylated poly-L-lysine, was evaluated further for the extent and specificity of receptor-mediated gene transfer into HuH-7 human hepatoma cells via the asialoglycoprotein receptor. Efficient receptor-mediated transfection occurred only when condensed DNA complexes had a spherical shape with a diameter of 15-30 nm; asialofetuin, a natural ligand for the asialoglycoprotein receptor, inhibited this process by up to 90%. Our results support the importance of appropriate DNA condensation for the uptake and ultimate expression of DNA in hepatic cells. PMID- 11438547 TI - Genetic deletion of the tumor necrosis factor receptor p60 or p80 abrogates ligand-mediated activation of nuclear factor-kappa B and of mitogen-activated protein kinases in macrophages. AB - Tumor necrosis factor (TNF) is a pleiotropic cytokine known to regulate cell growth, viral replication, inflammation, immune system functioning, angiogenesis, and tumorigenesis. These effects are mediated through two different receptors, TNFR1 and TNFR2 (also called p60 and p80, respectively), with p60 receptor being expressed on all cell types and p80 receptor only on cells of the immune system and on endothelial cells. Although the role of p60 receptor in TNF signaling is well established, the role of p80 is less clear. In this report, by using macrophages derived from wild-type mice (having both receptors) and mice in which the gene for either p60 (p60(-/-)), or p80 (p80(-/-)), or both (p60(-/-) p80(-/ )) receptor have been deleted, we have redefined the role of these receptors in TNF-induced activation of nuclear factor (NF)-kappa B and of mitogen-activated protein kinases. TNF activated NF-kappa B in a dose- and time-dependent manner in wild-type macrophages but not in p60(-/-), p80(-/-), or p60(-/-) p80(-/-) macrophages. These results correlated with the I kappa B alpha degradation needed for NF-kappa B activation. We also found that TNF activated c-Jun N-terminal protein kinase in a dose- and time-dependent manner in wild-type macrophages but not in p60(-/-), p80(-/-), or p60(-/-) p80(-/-) macrophages. TNF activated p38 MAPK and p44/p42 MAPK in wild-type but not in p60(-/-), p80(-/-), or p60(-/-) p80(-/-) macrophages. TNF induced the proliferation of wild-type macrophages, but for p60(-/-) and p80(-/-) macrophages proliferation was lower, and in p60(-/-) p80(-/-) it was absent. Overall, our studies suggest that both types of TNF receptors are needed in macrophages for optimum TNF cell signaling. PMID- 11438548 TI - Effects of tissue transglutaminase on retinoic acid-induced cellular differentiation and protection against apoptosis. AB - Retinoic acid (RA) and its various synthetic analogs affect mammalian cell growth, differentiation, and apoptosis. Whereas treatment of the human leukemia cell line HL60 with RA results in cellular differentiation, addition of the synthetic retinoid, N-(4-hydroxyphenyl) retinamide (HPR), induces HL60 cells to undergo apoptosis. Moreover, pretreatment of HL60 cells as well as other cell lines (i.e. NIH3T3 cells) with RA blocks HPR-induced cell death. In attempting to discover the underlying biochemical activities that might account for these cellular effects, we found that monodansylcadaverine (MDC), which binds to the enzyme (transamidase) active site of tissue transglutaminase (TGase), eliminated RA protection against cell death and in fact caused RA to become an apoptotic factor, suggesting that the ability of RA to protect against apoptosis is linked to the expression of active TGase. Furthermore, it was determined that expression of exogenous TGase in cells exhibited enhanced GTP binding and transamidation activities and mimicked the survival advantage imparted by RA. We tested whether the ability of this dual function enzyme to limit HPR-mediated apoptosis was a result of the ability of TGase to bind GTP and/or catalyze transamidation and found that GTP binding was sufficient for the protective effect. Moreover, excessive transamidation activity did not appear to be detrimental to cell viability. These findings, taken together with observations that the TGase is frequently up-regulated by environmental stresses, suggest that TGase may function to ensure cell survival under conditions of differentiation and cell stress. PMID- 11438549 TI - Homodimerization of amyloid precursor protein and its implication in the amyloidogenic pathway of Alzheimer's disease. AB - We reported previously that the carbohydrate domain of the amyloid precursor protein is involved in amyloid precursor protein (APP)-APP interactions. Functional in vitro studies suggested that this interaction occurs through the collagen binding site of APP. The physiological significance remained unknown, because it is not understood whether and how APP dimerization occurs in vivo. Here we report that cellular APP exists as homodimers matching best with a two site model. Consistent with our published crystallographic data, we show that a deletion of the entire sequence after the kunitz protease inhibitor domain did not abolish APP homodimerization, suggesting that two domains are critically involved but that neither is essential for homodimerization. Finally, we generated stabilized dimers by expressing mutant APP with a single cysteine in the ectodomain juxtamembrane region. Mutation of Lys(624) to cysteine produced approximately 6-8-fold more A beta than cells expressing normal APP. Our results suggest that amyloid A beta production can in principle be positively regulated by dimerization in vivo. We suggest that dimerization could be a physiologically important mechanism for regulating the proposed signal activity of APP. PMID- 11438550 TI - Akt inhibits the orphan nuclear receptor Nur77 and T-cell apoptosis. AB - Akt is a common mediator of cell survival in a variety of circumstances. Although some candidate Akt targets have been described, the function of Akt is not fully understood, particularly because of the cell type- and context-dependent apoptosis regulation. In this study, we demonstrate that one of the mechanisms by which Akt antagonizes apoptosis involves the inhibition of Nur77, a transcription factor implicated in T-cell receptor-mediated apoptosis. It has been suggested that Akt phosphorylates Nur77 directly, but whether Akt suppresses biological functions of Nur77 remains unknown. We found that Akt inhibited the DNA binding activity of Nur77 and stimulated its association with 14-3-3 in a phosphorylation site-dependent manner. Moreover, we found that expression of Akt suppressed Nur77 induced apoptosis in fibroblasts and activation-induced cell death of T-cell hybridomas. The inhibition of Nur77 by Akt suggests a mechanism that explains how T-cell receptor activation can promote survival in some instances even when Nur77 is induced. Collectively, these results may suggest that Akt is a negative regulator of Nur77 in T-cell apoptosis. PMID- 11438551 TI - Importance of Thr-353 of the conserved phosphorylation loop of the sarcoplasmic reticulum Ca2+-ATPase in MgATP binding and catalytic activity. AB - Mutants in which Thr-353 of the Ca(2+)-ATPase of sarcoplasmic reticulum had been replaced with alanine, serine, glutamine, cysteine, valine, aspartate, or tyrosine were analyzed functionally. All the mutations severely affected MgATP binding, whereas ATP binding was close to normal in the alanine, serine, glutamine, and valine mutants. In the serine and valine mutants, the maximum rate of phosphorylation from MgATP was 8- and 600-fold lower, respectively, compared with wild type. Replacement of Mg(2+) with Mn(2+) led to a 1.5-fold enhancement of the maximum phosphorylation rate in the valine mutant and a 5-fold reduction in the wild type. The turnover of the phosphoenzyme formed from MgATP was slowed 1-2 orders of magnitude relative to wild type in the alanine, serine, and valine mutants, but was close to normal in the aspartate and cysteine mutants. Only the serine mutant formed a phosphoenzyme in the backward reaction with P(i), and the hydrolysis of this intermediate was greatly enhanced. Analysis of the functional changes in the mutants in the light of the recent high resolution structure of the Ca(2+)-ATPase crystallized without the MgATP substrate suggests that, in the native activated state of the enzyme, the side chain hydroxyl of Thr-353 participates in important interactions with nucleotide and phosphate, possibly in catalysis, whereas the main chain carbonyl of Thr-353, but not the side chain, may coordinate the catalytic Mg(2+). PMID- 11438552 TI - Hsp90 phosphorylation is linked to its chaperoning function. Assembly of the reovirus cell attachment protein. AB - Studies on Hsp90 have mainly focused on its involvement in the activation of several families of protein kinases and of steroid hormone receptors. Little is known regarding the role of Hsp90 in the folding of nascent proteins. We previously reported that Hsp90 plays an active role in the posttranslational assembly of the C-terminal globular head of the reovirus attachment protein final sigma1. We show here that Hsp90 becomes phosphorylated in this process. However, only the unphosphorylated form of Hsp90 is complexed with final sigma1, suggesting that Hsp90 phosphorylation is coupled to the release of the chaperone from the target protein. Geldanamycin, which blocks final sigma1 maturation by preventing the release of Hsp90 from final sigma1, also inhibits Hsp90 phosphorylation. Taken together, these results demonstrate that Hsp90 phosphorylation is linked to its chaperoning function. PMID- 11438553 TI - Tamoxifen and contralateral breast cancer: the other side. PMID- 11438554 TI - Gene therapy for antiangiogenesis. PMID- 11438555 TI - Experts question validity of PSA testing for life insurance policies. PMID- 11438556 TI - STI571 revolution: can the newer targeted drugs measure up? PMID- 11438558 TI - After 30 years of laboratory work, a quick approval for STI571. PMID- 11438559 TI - Bladder cancer chemotherapy trial generates more questions than it answers. PMID- 11438560 TI - Biomarkers, animal models key in progress of chemoprevention. PMID- 11438561 TI - Jellyfish protein gives new glow to tumor imaging. PMID- 11438563 TI - National Institutes of Health Consensus Development Conference Statement: adjuvant therapy for breast cancer, November 1-3, 2000. AB - OBJECTIVE: Our goal was to provide health-care providers, patients, and the general public with an assessment of currently available data regarding the use of adjuvant therapy for breast cancer. PARTICIPANTS: The participants included a non-Federal, non-advocate, 14-member panel representing the fields of oncology, radiology, surgery, pathology, statistics, public health, and health policy as well as patient representatives. In addition, 30 experts in medical oncology, radiation oncology, biostatistics, epidemiology, surgical oncology, and clinical trials presented data to the panel and to a conference audience of 1000. EVIDENCE: The literature was searched with the use of MEDLINE(TM) for January 1995 through July 2000, and an extensive bibliography of 2230 references was provided to the panel. Experts prepared abstracts for their conference presentations with relevant citations from the literature. Evidence from randomized clinical trials and evidence from prospective studies were given precedence over clinical anecdotal experience. CONSENSUS PROCESS: The panel, answering predefined questions, developed its conclusions based on the evidence presented in open forum and the scientific literature. The panel composed a draft statement, which was read in its entirety and circulated to the experts and the audience for comment. Thereafter, the panel resolved conflicting recommendations and released a revised statement at the end of the conference. The panel finalized the revisions within a few weeks after the conference. The draft statement was made available on the World Wide Web immediately after its release at the conference and was updated with the panel's final revisions. The statement is available at http://consensus.nih.gov. CONCLUSIONS: The panel concludes that decisions regarding adjuvant hormonal therapy should be based on the presence of hormone receptor protein in tumor tissues. Adjuvant hormonal therapy should be offered only to women whose tumors express hormone receptor protein. Because adjuvant polychemotherapy improves survival, it should be recommended to the majority of women with localized breast cancer regardless of lymph node, menopausal, or hormone receptor status. The inclusion of anthracyclines in adjuvant chemotherapy regimens produces a small but statistically significant improvement in survival over non-anthracycline-containing regimens. Available data are currently inconclusive regarding the use of taxanes in adjuvant treatment of lymph node-positive breast cancer. The use of adjuvant dose intensive chemotherapy regimens in high-risk breast cancer and of taxanes in lymph node-negative breast cancer should be restricted to randomized trials. Ongoing studies evaluating these treatment strategies should be supported to determine if such strategies have a role in adjuvant treatment. Studies to date have included few patients older than 70 years. There is a critical need for trials to evaluate the role of adjuvant chemotherapy in these women. There is evidence that women with a high risk of locoregional tumor recurrence after mastectomy benefit from postoperative radiotherapy. This high-risk group includes women with four or more positive lymph nodes or an advanced primary cancer. Currently, the role of postmastectomy radiotherapy for patients with one to three positive lymph nodes remains uncertain and should be tested in a randomized controlled trial. Individual patients differ in the importance they place on the risks and benefits of adjuvant treatments. Quality of life needs to be evaluated in selected randomized clinical trials to examine the impact of the major acute and long-term side effects of adjuvant treatments, particularly premature menopause, weight gain, mild memory loss, and fatigue. Methods to support shared decision-making between patients and their physicians have been successful in trials; they need to be tailored for diverse populations and should be tested for broader dissemination. PMID- 11438564 TI - Effects of dietary N-(4-hydroxyphenyl)retinamide on N-nitrosomethylbenzylamine metabolism and esophageal tumorigenesis in the Fischer 344 rat. AB - BACKGROUND: 9-cis-Retinoic acid (9-cis-RA) and N-(4-hydroxyphenyl)retinamide (4 HPR) are effective chemopreventive agents against epithelial tumors in the oral cavity, breast, and prostate. We tested the inhibitory activity of these retinoids against N-nitrosomethylbenzylamine (NMBA)-induced tumorigenesis in the rat esophagus. METHODS: Male Fischer 344 rats were randomly assigned to receive diets either lacking or containing 9-cis-RA or 4-HPR for 1 week before tumor initiation with NMBA and then for the duration of the study. NMBA metabolism, O(6)-methylguanine adduct formation, and cytochrome P450 messenger RNA (mRNA) expression in the esophagi of the rats were studied to investigate the mechanisms by which dietary 4-HPR affects tumorigenesis. All statistical tests were two sided. RESULTS: Dietary 4-HPR resulted in a dose-dependent and statistically significant enhancement (P<.05) of tumorigenesis in response to NMBA. In two different tumor bioassays, the mean tumor multiplicity for rats fed the highest concentration of dietary 4-HPR (0.8 g/kg diet) was increased by 5.9 tumors (95% confidence interval [CI] = 1.7 to 10.1 tumors) and 6.7 tumors (95% CI = 5.6 to 7.8 tumors) compared with the mean tumor multiplicity for rats that received the control diet lacking 4-HPR. Animals fed diets containing 9-cis-RA displayed no statistically significant increase in tumorigenesis. Compared with animals fed a diet lacking 4-HPR, animals fed 4-HPR had increased NMBA metabolism in esophageal explant cultures and had higher levels of O(6)-methylguanine DNA adducts and CYP2A3 mRNA in their esophagi. CONCLUSIONS: Dietary 4-HPR enhances tumorigenesis in response to NMBA in the rat esophagus by increasing tumor initiation events. Dietary 4-HPR may exert paradoxical effects at some sites, such as the aerodigestive tract, by modulating the bioactivation of carcinogens in target tissues. PMID- 11438565 TI - Inhibition of murine prostate tumor growth and activation of immunoregulatory cells with recombinant canarypox viruses. AB - BACKGROUND: Immunization with modified tumor cells carrying recombinant immunomodulatory genes is being explored as cancer immunotherapy. In this study, we examine whether canarypox ALVAC viruses carrying immunostimulatory cytokine genes (granulocyte-macrophage colony-stimulating factor, interleukin 2, interleukin 12, and tumor necrosis factor-alpha) can induce antitumor immunity (to rechallenge) in the RM-1 model of a highly aggressive, weakly immunogenic murine prostate cancer. METHODS: For antitumor activity studies, RM-1 murine prostate cancer cells were infected with the parental ALVAC virus or one or two recombinant ALVAC-cytokine viruses and then injected into male C57BL/6 mice. For rechallenge studies, other mice were first given an injection subcutaneously with irradiated (nonproliferating) recombinant ALVAC-infected RM-1 cells and then (10 days later) with untreated RM-1 cells. For the determination of which immune cells were required for antitumor activity, mice were immunodepleted of CD4, CD8, or natural killer (NK) NK1.1 cells with the corresponding monoclonal antibodies and were then given an injection of ALVAC-cytokine-infected RM-1 cells. For all experiments, tumor outgrowth and animal survival were monitored. RESULTS: After subcutaneous injection into mice, RM-1 cells infected with one (except ALVAC interleukin 2) or two ALVAC-cytokine recombinants had statistically significantly greater antitumor activity than RM-1 cells infected with parental ALVAC (P<.001 for all; two-sided test). The antitumor activity of RM-1 cells infected with any two ALVAC-cytokine recombinants was greater than, but not statistically significantly different from, that of RM-1 cells infected with any one ALVAC cytokine recombinant. NK1.1 cells were necessary for antitumor activity, but tumor-specific CD4(+) regulatory T cells were also induced that inhibited CD8(+) RM-1-specific cytotoxic T cells, resulting in the lack of immunity to a rechallenge by RM-1 cells. DISCUSSION: Canarypox viruses can transfer immunostimulatory cytokine genes into RM-1 prostate cancer cells. When such cells were injected into mice, the cytokines induced an antitumor response against this highly aggressive, weakly immunogenic tumor. This response, however, did not protect the mouse against a rechallenge with RM-1 cells because suppressor CD4(+) T cells were induced that inhibited tumor-specific CD8(+) cytotoxic T cells. PMID- 11438566 TI - Tamoxifen therapy for primary breast cancer and risk of contralateral breast cancer. AB - BACKGROUND: Women diagnosed with breast cancer have a twofold to sixfold greater risk of developing contralateral breast cancer than women in the general population have of developing a first breast cancer. Tamoxifen therapy reduces this risk, but it is unclear if this benefit exists for both estrogen receptor (ER)-positive and ER-negative contralateral tumors. METHODS: Using data from a population-based tumor registry that collects information on the ER status of breast tumors, we followed 8981 women residing in western Washington State who were diagnosed with a primary unilateral invasive breast cancer during the period from 1990 through 1998 to identify cases of contralateral breast cancer. We restricted our analyses to women who were at least 50 years old and whose first breast cancer had a localized or regional stage; women who received adjuvant hormonal therapy but not chemotherapy (n = 4654) were classified as tamoxifen users, while those who received neither adjuvant hormonal therapy nor chemotherapy (n = 4327) were classified as nonusers of tamoxifen. By reviewing selected patient abstracts, we estimated that 94% of the subjects were classified correctly with respect to tamoxifen use. The risk of contralateral breast cancer associated with tamoxifen use was estimated with the use of Cox regression. All statistical tests were two-sided. RESULTS: Of the 89 tamoxifen users and 100 nonusers of tamoxifen diagnosed with contralateral breast cancer, 112 had ER positive tumors, 20 had ER-negative tumors, and 57 had tumors with an ER status that was unknown or had not been determined by an immunohistochemical assay. The risk of developing an ER-positive and an ER-negative contralateral tumor among tamoxifen users was 0.8 (95% confidence interval [CI] = 0.5 to 1.1) and 4.9 (95% CI = 1.4 to 17.4), respectively, times that of nonusers of tamoxifen. This difference in risk by ER status was statistically significant (P<.0001). CONCLUSIONS: Tamoxifen use appears to decrease the risk of ER-positive contralateral breast tumors, but it appears to increase the risk of ER-negative contralateral tumors. PMID- 11438567 TI - Effect of retroviral endostatin gene transfer on subcutaneous and intraperitoneal growth of murine tumors. AB - BACKGROUND: Inhibiting tumor angiogenesis is a promising new strategy for treating cancer. Difficulties with the stability, manufacture, and long-term administration of recombinant antiangiogenic proteins have prompted investigators to use gene therapy to generate these proteins in vivo. We investigated whether transfer of the gene encoding the angiogenesis inhibitor endostatin into the murine liver cell line NMuLi could inhibit tumor growth in vivo. METHODS: NMuLi cells were transduced with retroviral vectors containing the murine endostatin gene. The presence and function of endostatin in transduced cell supernatants were confirmed by competitive enzyme immunoassay and endothelial cell proliferation assays. Nude mice were given a subcutaneous or intraperitoneal injection with NMuLi cells, control transduced cells (NEF-null), or endostatin transduced clones (NEF-Endo1 to 4) and were monitored for tumor growth. All statistical tests were two-sided. RESULTS: Supernatants from the clone secreting the lowest amount of endostatin (NEF-Endo4, 28 ng/mL) inhibited endothelial cell proliferation by 6% (95% confidence interval [CI] = 0% to 12%), and those from the clone secreting the highest amount (NEF-Endo1, 223 ng/mL) inhibited endothelial cell proliferation by 20% (95% CI = 13% to 27%). Increased levels of endostatin were detected in tumor lysates, but not serum, of mice given a subcutaneous injection of NEF-Endo1 cells. After 63 days, mice given a subcutaneous injection of parental NMuLi or NEF-null cells had tumor volumes of 2400 mm(3) (95% CI = 1478 mm(3) to 3300 mm(3)) and 2700 mm(3) (95% CI = 2241 mm(3) to 3144 mm(3)), respectively, compared with mean tumor volumes of less than 30 mm(3) in mice given an injection of NEF-Endo clones, a statistically significant difference (P<.001). After 123 days, all 16 mice given an intraperitoneal injection of parental NMuLi or NEF-null cells had died, compared with only three (9%) of 32 mice given an injection of NEF-Endo clones. CONCLUSIONS: Retroviral endostatin gene transfer leads to secretion of functional endostatin that is sufficiently active to inhibit tumor growth. Further studies of retroviral endostatin gene transfer for the treatment of cancer are warranted. PMID- 11438568 TI - Cancer mortality after nasopharyngeal radium irradiation in the Netherlands: a cohort study. AB - BACKGROUND: Nasopharyngeal radium irradiation (NRI) was used widely from 1940 through 1970 to treat otitis serosa in children and barotrauma in airmen and submariners. We assessed whether NRI-exposed individuals were at higher risk for cancer-related deaths than were nonexposed individuals. METHODS: We conducted a retrospective cohort study of all-cause and cancer-related mortality in 5358 NRI exposed subjects and in 5265 frequency-matched nonexposed subjects, who as children were treated at nine ear, nose, and throat clinics in The Netherlands from 1945 through 1981. We recorded personal and medical data from original patient medical records and assessed vital status through follow-up at municipal population registries. Risk of mortality was evaluated by standardized mortality ratios (SMRs). All statistical tests were two-sided. RESULTS: The average radiation doses were 275, 10.9, 1.8, and 1.5 cGy for the nasopharynx, pituitary, brain, and thyroid, respectively. The median follow-up was 31.6 years. Three hundred two NRI-exposed subjects had died, with 269.2 deaths expected (SMR = 1.1; 95% confidence interval [CI] = 1.0 to 1.3); among nonexposed subjects, 315 died, with 283.5 deaths expected (SMR = 1.1; 95% CI = 0.99 to 1.2). Cancer-related deaths of 96 exposed subjects (SMR = 1.2; 95% CI = 0.95 to 1.4) and 87 nonexposed subjects (SMR = 1.0; 95% CI = 0.8 to 1.3) were documented. There were no excess deaths from cancers of the head and neck area among exposed subjects. However, there were excess deaths from cancers of lymphoproliferative and hematopoietic origin (SMR = 1.9; 95% CI = 1.1 to 3.0), mainly from non-Hodgkin's lymphoma (SMR = 2.6; 95% CI = 1.0 to 5.3). We found no evidence that breast cancer deaths were less than expected (SMR = 1.7; 95% CI = 0.9 to 2.8) in contrast to an earlier study. CONCLUSIONS: Our findings do not indicate an increased cancer mortality risk in a population exposed to NRI in childhood. More prolonged follow-up of this and other NRI cohorts is recommended. PMID- 11438569 TI - G(alpha)q-deficient mice lack metabotropic glutamate receptor-dependent long-term depression but show normal long-term potentiation in the hippocampal CA1 region. AB - Long-term potentiation (LTP) and depression (LTD) are potential cellular mechanisms involved in learning and memory. Group I metabotropic glutamate receptors (mGluR), which are linked to heterotrimeric G-proteins of the G(q) family (G(q) and G(11)), have been reported to facilitate both hippocampal LTP and LTD. To evaluate their functional role in synaptic plasticity, we studied LTD and LTP in the CA1 region of the hippocampus from wild-type, Galpha(q)(-/-), and Galpha(11)(-/-) mice. Basic parameters of the synaptic transmission were not altered in Galpha(q)(-/-) and Galpha(11)(-/-) mice. Moreover, these mice showed normal LTP in response to a strong tetanus and to a weak tetanus. However, LTD induced either by a group I mGluRs agonist or by paired-pulse low-frequency stimulation (PP-LFS) was absent in Galpha(q)(-/-) mice. Moreover, PP-LFS caused potentiation of the synaptic transmission in these mice that was not affected by the NMDAR antagonist AP-5. These results show that G(q) plays a crucial role in the mGluR-dependent LTD, whereas hippocampal LTP is not affected by the lack of a single member of the G(q) family. PMID- 11438570 TI - Evidence for a role of mixed lineage kinases in neuronal apoptosis. AB - Superior cervical ganglion (SCG) sympathetic neurons die by apoptosis when deprived of nerve growth factor (NGF). It has been shown previously that the induction of apoptosis in these neurons at NGF withdrawal requires both the activity of the small GTP-binding protein Cdc42 and the activation of the c-Jun N terminal kinase (JNK) pathway. The mixed lineage kinase 3 (MLK3) belongs to a family of mitogen-activated protein (MAP) kinase kinase kinases. MLK3 contains a Cdc42/Rac interactive-binding (CRIB) domain and activates both the JNK and the p38 MAP kinase pathways. In this study the role of MLK3 in the induction of apoptosis in sympathetic neurons has been investigated. Overexpression of an active MLK3 induces activation of the JNK pathway and apoptosis in SCG neurons. In addition, overexpression of kinase dead mutants of MLK3 blocks apoptosis as well as c-Jun phosphorylation induced by NGF deprivation. More importantly, MLK3 activity seems to increase by 5 hr after NGF withdrawal in both differentiated PC12 cells and SCG neurons. We also show that MLK3 lies downstream of Cdc42 in the neuronal death pathway. Regulation of MLK3 in neurons seems to be dependent on MLK3 activity and possibly on an additional cellular component, but not on its binding to Cdc42. These results suggest that MLK3, or a closely related kinase, is a physiological element of NGF withdrawal-induced activation of the Cdc42-c Jun pathway and neuronal death. MLK3 therefore could be an interesting therapeutic target in a number of neurodegenerative diseases involving neuronal apoptosis. PMID- 11438571 TI - Negative cross talk between anionic GABAA and cationic P2X ionotropic receptors of rat dorsal root ganglion neurons. AB - Using whole-cell patch-clamp recording and intracellular Ca(2+) imaging of rat cultured DRG neurons, we studied the cross talk between GABA(A) and P2X receptors. A rapidly fading current was the main response to ATP, whereas GABA elicited slowly desensitizing inward currents. Coapplication of these agonists produced a total current much smaller than the linear summation of individual responses (68 +/- 5% with 10 microm ATP plus 100 microm GABA). Occlusion was observed regardless of ATP response type. Neurons without functional P2X receptors manifested no effect of ATP on GABA currents (and vice versa). Occlusion was also absent in the presence of the P2X blocker trinitrophenyl-ATP (TNP-ATP) or of the GABA blocker picrotoxin, indicating a lack of involvement by metabotropic ATP or GABA receptors. Less occlusion was obtained when ATP was applied 2 sec after GABA than when GABA was applied after ATP. Changing the polarity of GABA currents by using intracellular SO(4)2- instead of Cl(-) significantly reduced the occlusion of ATP currents by GABA, suggesting an important role for Cl(-) efflux in this phenomenon. Occlusion was enhanced whenever intracellular Ca(2+) ([Ca(2+)](i)) was not buffered, indicating the cross talk-facilitating role of this divalent cation. Ca(2+) imaging showed that ATP (but not GABA) increased [Ca(2+)](i) in voltage-clamped or intact neurons. Our data demonstrated a novel Cl(-) and Ca(2+)-dependent interaction between cationic P2X and anionic GABA(A) receptors of DRG neurons. Such negative cross talk might represent a model for a new mechanism to inhibit afferent excitation to the spinal cord as GABA and ATP are coreleased within the dorsal horn. PMID- 11438572 TI - Predominance of late-spiking neurons in layer VI of rat perirhinal cortex. AB - Recent work demonstrated the importance of perirhinal cortex (PR) in a variety of behavioral tasks and disease processes. Studies from our laboratory revealed that some layers of PR contain neurons with unusual properties. Here we report a detailed examination of the cellular neurobiology of layer VI of PR, using whole cell recordings and biocytin cell fills in horizontal rat brain slices. The most striking finding is that an overwhelming majority ( approximately 86%) of neurons are late-spiking (LS) cells, which can delay the onset of their spike trains by several seconds or more relative to the onset of a depolarizing current step. LS neurons previously have been shown to exist only in very small numbers in a limited number of other cortical regions. Anatomical reconstructions have revealed that the LS neurons vary greatly in morphology, including both pyramidal and nonpyramidal cells. Another surprising physiological finding is the fact that single-spiking (SS) neurons are the second most common cell type ( approximately 7%). SS neurons issue only a single action potential even in response to extreme depolarization. They have been seen previously in the amygdala, but never in cortex. A third remarkable finding is that there are almost no regular spiking (RS) neurons, unlike all other cortical regions that have been studied. This unique abundance of LS neurons in layer VI, along with the presence of SS neurons and the absence of RS neurons, demonstrates that layer VI of PR is unlike any other cortical region that has been studied to date. PMID- 11438573 TI - A (beta)-strand in the (gamma)2 subunit lines the benzodiazepine binding site of the GABA A receptor: structural rearrangements detected during channel gating. AB - Benzodiazepines (BZDs) exert their effects in the CNS by binding to a modulatory site on GABA(A) receptors. Individual amino acids have been implicated in BZD recognition and modulation of the GABA(A) receptor, but the secondary structure of the amino acids contributing to the BZD binding site has not been elucidated. In this report we used the substituted cysteine accessibility method to understand the structural dynamics of a region of the GABA(A) receptor implicated in BZD binding, gamma(2)Y72-gamma(2)Y83. Each residue within this region was mutated to cysteine and expressed with wild-type alpha(1) and beta(2) subunits in Xenopus oocytes. Methanethiosulfonate (MTS) reagents were used to modify covalently the engineered cysteines, and the subsequent effects on BZD modulation of the receptor were monitored functionally by two-electrode voltage clamp. We identified an alternating pattern of accessibility to sulfhydryl modification, indicating that the region gamma(2)T73-gamma(2)T81 adopts a beta-strand conformation. By monitoring the ability of BZD ligands to impede the covalent modification of accessible cysteines, we also identified two residues within this region, gamma(2)A79 and gamma(2)T81, that line the BZD binding site. Sulfhydryl modification of gamma(2)A79C or gamma(2)T81C allosterically shifts the GABA EC(50) of the receptor, suggesting that certain MTS compounds may act as tethered agonists at the BZD binding site. Last, we present structural evidence that a portion of the BZD binding site undergoes a conformational change in response to GABA binding and channel gating (opening and desensitization). These data represent an important step in understanding allosteric communication in ligand gated ion channels. PMID- 11438574 TI - Dishevelled regulates the metabolism of amyloid precursor protein via protein kinase C/mitogen-activated protein kinase and c-Jun terminal kinase. AB - Alzheimer's disease (AD) is a disorder of two pathologies: amyloid plaques, the core of which is a peptide derived from the amyloid precursor protein (APP), and neurofibrillary tangles composed of highly phosphorylated tau. Protein kinase C (PKC) is known to increase non-amyloidogenic alpha-secretase cleavage of APP, producing secreted APP (sAPPalpha), and glycogen synthase kinase (GSK)-3beta is known to increase tau phosphorylation. Both PKC and GSK-3beta are components of the wnt signaling cascade. Here we demonstrate that overexpression of another member of this pathway, dishevelled (dvl-1), increases sAPPalpha production. The dishevelled action on APP is mediated via both c-jun terminal kinase (JNK) and protein kinase C (PKC)/mitogen-activated protein (MAP) kinase but not via p38 MAP kinase. These data position dvl-1 upstream of both PKC and JNK, thereby explaining the previously observed dual signaling action of dvl-1. Furthermore, we show that human dvl-1 and wnt-1 also reduce the phosphorylation of tau by GSK 3beta. Therefore, both APP metabolism and tau phosphorylation are potentially linked through wnt signaling. PMID- 11438575 TI - FGF induces a switch in death receptor pathways in neuronal cells. AB - Basic fibroblast growth factor (FGF2) has many roles in neuronal development and maintenance including effects on mitogenesis, survival, fate determination, differentiation, and migration. Using a conditionally immortalized rat hippocampal cell line, H19-7, and primary hippocampal cultures, we now demonstrate that FGF2 treatment differentially regulates members of the tumor necrosis factor (TNF) superfamily of death domain receptors and their ligands. H19-7 cells transferred from serum to defined (N2) medium undergo apoptosis by a Fas-dependent mechanism similar to primary neurons. In contrast, H19-7 cells treated with FGF undergo apoptosis by a Fas-independent mechanism. FGF suppresses the Fas death pathway but also induces apoptosis by activation of a TNFalpha death pathway in both H19-7 and hippocampal progenitor cells. Expression of the TNF receptor 1 (TNFR1) or TNFR2 in H19-7 cells is sufficient to sensitize the cells to TNFalpha, similar to the effects of FGF. Because TNFalpha can be either proapoptotic or antiapoptotic, these results provide an explanation for the divergent trophic effects of FGF2 treatment and the observation that multiple trophic inputs are required for the survival of specific neurons. PMID- 11438576 TI - Death in the balance: alternative participation of the caspase-2 and -9 pathways in neuronal death induced by nerve growth factor deprivation. AB - The data presented here demonstrate that sympathetic neurons have the potential to activate two alternative caspase-dependent pathways either of which is capable of mediating death induced by NGF deprivation and that these neurons have the potential to switch from one pathway to the other. The presence of these two alternative pathways to trophic factor deprivation-induced death may have implications for ensuring the correct development of the nervous system. In wild type neurons, a caspase-2-dependent pathway is required for death, and a caspase 9-dependent pathway appears to be suppressed by endogenous inhibitors of apoptosis proteins (IAPs). In contrast, for caspase-2-null neurons, death is dependent on the caspase-9 pathway. The mechanism underlying the shift is the result of a threefold compensatory elevation of caspase-9 expression and a doubling of levels of direct IAP binding protein with low pI/(DIABLO)/second mitochondria-derived activator of caspase (Smac), an IAP inhibitor, both at the mRNA and protein levels [corrected]. These findings resolve seemingly discrepant findings regarding the roles of various caspases after NGF deprivation and raise a cautionary note regarding the interpretation of findings with caspase-null animals. The choice of the death-mediating caspase pathway in the sympathetic neurons is thus dependent on the regulated relative expression of components of the pathways including those of caspases, IAPs, and IAP inhibitors. PMID- 11438577 TI - Cyclin-dependent kinases and P53 pathways are activated independently and mediate Bax activation in neurons after DNA damage. AB - DNA damage has been implicated as one important initiator of cell death in neuropathological conditions such as stroke. Accordingly, it is important to understand the signaling processes that control neuronal death induced by this stimulus. Previous evidence has shown that the death of embryonic cortical neurons treated with the DNA-damaging agent camptothecin is dependent on the tumor suppressor p53 and cyclin-dependent kinase (CDK) activity and that the inhibition of either pathway alone leads to enhanced and prolonged survival. We presently show that p53 and CDKs are activated independently on parallel pathways. An increase in p53 protein levels, nuclear localization, and DNA binding that result from DNA damage are not affected by the inhibition of CDK activity. Conversely, no decrease in retinoblastoma protein (pRb) phosphorylation was observed in p53-deficient neurons that were treated with camptothecin. However, either p53 deficiency or the inhibition of CDK activity alone inhibited Bax translocation, cytochrome c release, and caspase-3-like activation. Taken together, our results indicate that p53 and CDK are activated independently and then act in concert to control Bax-mediated apoptosis. PMID- 11438578 TI - Chemokines and glycoprotein120 produce pain hypersensitivity by directly exciting primary nociceptive neurons. AB - Human immunodeficiency virus-1 (HIV-1) infection is associated with numerous effects on the nervous system, including pain and peripheral neuropathies. We now demonstrate that cultured rat dorsal root ganglion (DRG) neurons express a wide variety of chemokine receptors, including those that are thought to act as receptors for the HIV-1 coat protein glycoprotein120 (gp120). Chemokines that activate all of the known chemokine receptors increased [Ca(2+)](i) in subsets of cultured DRG cells. Many neurons responded to multiple chemokines and also to bradykinin, ATP, and capsaicin. Immunohistochemical studies demonstrated the expression of the CXCR4 and CCR4 chemokine receptors on populations of DRG neurons that also expressed substance P and the VR1 vanilloid receptor. RT-PCR analysis confirmed the expression of CXCR4, CX3CR1, CCR4, and CCR5 mRNAs in DRG neurons. Chemokines and gp120 produced excitatory effects on DRG neurons and also stimulated the release of substance P. Chemokines and gp120 also produced allodynia after injection into the rat paw. Thus these results provide evidence that chemokines and gp120 may produce painful effects via direct actions on chemokine receptors expressed by nociceptive neurons. Chemokine receptor antagonists may be important therapeutic interventions in the pain that is associated with HIV-1 infection and inflammation. PMID- 11438579 TI - Dynamics of glycine receptor insertion in the neuronal plasma membrane. AB - The exocytosis site of newly synthesized glycine receptor was defined by means of a morphological assay to characterize its export from the trans-Golgi Network to the plasma membrane. This was achieved by expressing in transfected neurons an alpha1 subunit bearing an N-terminal tag selectively cleavable from outside the cell by thrombin. This was combined with a transient temperature-induced block of exocytic transport that creates a synchronized exocytic wave. Immunofluorescence microscopy analysis of the cell surface appearance of newly synthesized receptor revealed that exocytosis mainly occurred at nonsynaptic sites in the cell body and the initial portion of dendrites. At the time of cell surface insertion, the receptors existed as discrete clusters. Quantitative analysis showed that glycine receptor clusters are stable in size and subsequently appeared in more distal dendritic regions. This localization resulted from diffusion in the plasma membrane and not from exocytosis of transport vesicles directed to dendrites. Kinetic analysis established a direct substrate-product relationship between pools of somatic and dendritic receptors. This indicated that clusters represent intermediates between newly synthesized and synaptic receptors. These results support a diffusion-retention model for the formation of receptor-enriched postsynaptic domains and not that of a vectorial intracellular targeting to synapses. PMID- 11438580 TI - Mild cerebral ischemia induces loss of cyclin-dependent kinase inhibitors and activation of cell cycle machinery before delayed neuronal cell death. AB - After mild ischemic insults, many neurons undergo delayed neuronal death. Aberrant activation of the cell cycle machinery is thought to contribute to apoptosis in various conditions including ischemia. We demonstrate that loss of endogenous cyclin-dependent kinase (Cdk) inhibitor p16(INK4a) is an early and reliable indicator of delayed neuronal death in striatal neurons after mild cerebral ischemia in vivo. Loss of p27(Kip1), another Cdk inhibitor, precedes cell death in neocortical neurons subjected to oxygen-glucose deprivation in vitro. The loss of Cdk inhibitors is followed by upregulation of cyclin D1, activation of Cdk2, and subsequent cytoskeletal disintegration. Most neurons undergo cell death before entering S-phase, albeit a small number ( approximately 1%) do progress to the S-phase before their death. Treatment with Cdk inhibitors significantly reduces cell death in vitro. These results show that alteration of cell cycle regulatory mechanisms is a prelude to delayed neuronal death in focal cerebral ischemia and that pharmacological interventions aimed at neuroprotection may be usefully directed at cell cycle regulatory mechanisms. PMID- 11438581 TI - Spontaneous changes in mitochondrial membrane potential in cultured neurons. AB - Using the mitochondrial membrane potential (DeltaPsi(m))-sensitive fluorescent dyes 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine iodide (JC-1) and tetramethylrhodamine methyl ester (TMRM), we have observed spontaneous changes in the DeltaPsi(m) of cultured forebrain neurons. These fluctuations in DeltaPsi(m) appear to represent partial, transient depolarizations of individual mitochondria. The frequency of these DeltaPsi(m) fluctuations can be significantly lowered by exposure to a photo-induced oxidant burden, an ATP synthase inhibitor, or a glutamate-induced sodium load, without changing overall JC-1 fluorescence intensity. These spontaneous fluctuations in JC-1 signal were not inhibited by altering plasma membrane activity with tetrodotoxin or MK-801 or by blocking the mitochondrial permeability transition pore (PTP) with cyclosporin A. Neurons loaded with TMRM showed similar, low-amplitude, spontaneous fluctuations in DeltaPsi(m). We hypothesize that these DeltaPsi(m) fluctuations are dependent on the proper functioning of the mitochondria and reflect mitochondria alternating between the active and inactive states of oxidative phosphorylation. PMID- 11438582 TI - Plasma membrane Ca2+-ATPase isoform 2a is the PMCA of hair bundles. AB - Mechanoelectrical transduction channels of hair cells allow for the entry of appreciable amounts of Ca(2+), which regulates adaptation and triggers the mechanical activity of hair bundles. Most Ca(2+) that enters transduction channels is extruded by the plasma membrane Ca(2+)-ATPase (PMCA), a Ca(2+) pump that is highly concentrated in hair bundles and may be essential for normal hair cell function. Because PMCA isozymes and splice forms are regulated differentially and have distinct biochemical properties, we determined the identity of hair bundle PMCA in frog and rat hair cells. By screening a bullfrog saccular cDNA library, we identified abundant PMCA1b and PMCA2a clones as well as rare PMCA2b and PMCA2c clones. Using immunocytochemistry and immunoprecipitation experiments, we showed in bullfrog sacculus that PMCA1b is the major isozyme of hair cell and supporting cell basolateral membranes and that PMCA2a is the only PMCA present in hair bundles. This complete segregation of PMCA1 and PMCA2 isozymes holds for rat auditory and vestibular hair cells; PMCA2a is the only PMCA isoform in hair bundles of outer hair cells and vestibular hair cells and is the predominant PMCA of hair bundles of inner hair cells. Our data suggest that hair cells control plasma membrane Ca(2+)-pumping activity by targeting specific PMCA isozymes to distinct subcellular locations. Because PMCA2a is the only Ca(2+) pump present at appreciable levels in hair bundles, the biochemical properties of this pump must account fully for the physiological features of transmembrane Ca(2+) pumping in bundles. PMID- 11438583 TI - cAMP-dependent protein kinase mediates activity-regulated synaptic targeting of NMDA receptors. AB - Chronic activity blockade increases synaptic levels of NMDA receptor immunoreactivity in hippocampal neurons. We show here that blockade-induced synaptic NMDA receptors are functional and mediate enhanced excitotoxicity in response to synaptically released glutamate. Activity blockade increased the cell surface association of NMDA receptors. Blockade-induced synaptic targeting of NMDA receptors did not require protein synthesis but required phosphorylation and specifically cAMP-dependent protein kinase (PKA). Furthermore, activation of PKA was sufficient to induce synaptic targeting of NMDA receptors regardless of receptor activity status. These results implicate PKA activity downstream of receptor blockade as a mediator of enhanced synaptic transport or stabilization of NMDA receptors. Synaptic clustering of NR1-green fluorescent protein was observed in living neurons in response to NMDA receptor and cAMP phosphodiesterase antagonists and occurred gradually over the course of a day. This pathway represents a cellular mechanism for synaptic homeostasis and is likely to function in metaplasticity, long-term regulation of the ability of a synapse to undergo potentiation or depression. PMID- 11438584 TI - Experience-dependent gene expression in the rat hippocampus after spatial learning: a comparison of the immediate-early genes Arc, c-fos, and zif268. AB - Neuronal immediate-early gene (IEG) expression is regulated by synaptic activity and plays an important role in the neuroplastic mechanisms critical to memory consolidation. IEGs can be divided into two functional classes: (1) regulatory transcription factors (RTFs), which can broadly influence cell function depending on the "downstream" genes they regulate, and (2) "effector" proteins, which may directly modulate specific cellular functions. The objective of the current study was to determine whether the expression of an effector IEG (Arc) was similar to, or different from, that of two well characterized RTF IEGs (c-fos and zif268) after learning. IEG RNA levels from rats trained in spatial and nonspatial water tasks were determined using RNase protection assays and in situ hybridization. Overall, the regulation of the three IEGs was similar in the hippocampus and the entorhinal and primary visual cortices. Consequently, IEG RNA levels were positively correlated within a structure. By contrast, Arc and zif268 RNA levels were not correlated or only weakly correlated across structures, although c-fos RNA levels were moderately correlated across structures. Arc RNA expression differed from that of zif268 and c-fos in two regards: (1) hippocampal Arc RNA levels were correlated with learning of the hippocampal-dependent spatial, but not hippocampal-independent cued response, water task, and (2) Arc RNA levels in the hippocampus and entorhinal cortex increased after spatial reversal learning relative to an asymptotic performance group. Thus, although the expression of Arc, zif268, and c-fos exhibited many similarities, Arc was most responsive to differences in behavioral task demands. PMID- 11438585 TI - Genetic control of the mouse cerebellum: identification of quantitative trait loci modulating size and architecture. AB - To discover genes influencing cerebellum development, we conducted a complex trait analysis of variation in the size of the adult mouse cerebellum. We analyzed two sets of recombinant inbred BXD strains and an F2 intercross of the common inbred strains, C57BL/6J and DBA/2J. We measured cerebellar size as the weight or volume of fixed or histologically processed tissue. Among BXD recombinant inbred strains, the cerebellum averages 52 mg (12.4% of the brain) and ranges 18 mg in size. In F2 mice, the cerebellum averages 62 mg (12.9% of the brain) and ranges approximately 20 mg in size. Five quantitative trait loci (QTLs) that significantly control variation in cerebellar size were mapped to chromosomes 1 (Cbs1a), 8 (Cbs8a), 14 (Cbs14a), and 19 (Cbs19a, Cbs19b). In combination, these QTLs can shift cerebellar size an appreciable 35% of the observed range. To assess regional genetic control of the cerebellum, we also measured the volume of the cell-rich, internal granule layer (IGL) in a set of BXD strains. The IGL ranges from 34 to 43% of total cerebellar volume. The QTL Cbs8a is significantly linked to variation in IGL volume and is suggestively linked to variation in the number of cerebellar folia. The QTLs we have discovered are among the first loci shown to modulate the size and architecture of the adult mouse cerebellum. PMID- 11438586 TI - Inhibition of mitochondrial complex II induces a long-term potentiation of NMDA mediated synaptic excitation in the striatum requiring endogenous dopamine. AB - Abnormal involuntary movements and cognitive impairment represent the classical clinical symptoms of Huntington's disease (HD). This genetic disorder involves degeneration of striatal spiny neurons, but not striatal large cholinergic interneurons, and corresponds to a marked decrease in the activity of mitochondrial complex II [succinate dehydrogenase (SD)] in the brains of HD patients. Here we have examined the possibility that SD inhibitors exert their toxic action by increasing glutamatergic transmission. We report that SD inhibitors such as 3-nitroproprionic acid (3-NP), but not an inhibitor of mitochondrial complex I, produce a long-term potentiation of the NMDA-mediated synaptic excitation (3-NP-LTP) in striatal spiny neurons. In contrast, these inhibitors had no effect on excitatory synaptic transmission in striatal cholinergic interneurons and pyramidal cortical neurons. 3-NP-LTP involves increased intracellular calcium and activation of the mitogen-activated protein kinase extracellular signal-regulated kinase and is critically dependent on endogenous dopamine acting via D2 receptors, whereas it is negatively regulated by D1 receptors. Thus 3-NP-LTP might play a key role in the regional and cell type-specific neuronal death observed in HD. PMID- 11438587 TI - A novel p75NTR signaling pathway promotes survival, not death, of immunopurified neocortical subplate neurons. AB - Subplate neurons of mammalian neocortex undergo pronounced cell death postnatally, long after they have matured and become incorporated into functional cortical circuits. They express the p75 neurotrophin receptor (p75NTR), which is known to signal cell death in some types of neurons via the activation of sphingomyelinase and the concomitant increase in the sphingolipid ceramide. To evaluate the role of p75NTR in subplate neurons, they were immunopurified and cultured in vitro. Contrary to its known function as a death receptor, ligand binding to p75NTR promotes subplate neuron survival. Moreover, p75NTR-dependent survival is blocked by inhibition of ceramide synthesis and rescued by addition of its precursor sphingomyelin. Inhibition of Trk signaling does not block survival, nor is Trk signaling alone sufficient to promote survival. Thus, ligand dependent p75NTR regulation of the ceramide pathway mediates survival in certain neurons and may represent an important target for neuroprotective drugs in degenerative diseases involving p75NTR-expressing neurons, such as Alzheimer's disease. PMID- 11438588 TI - The leukocyte common antigen-related protein tyrosine phosphatase receptor regulates regenerative neurite outgrowth in vivo. AB - Drosophila and leech models of nervous system development demonstrate that protein tyrosine phosphatase (PTP) receptors regulate developmental neurite outgrowth. Whether PTP receptors regulate neurite outgrowth in adult systems or in regenerative states remains unknown. The leukocyte common antigen-related (LAR) receptor is known to be present in rodent dorsal root ganglion (DRG) neurons; therefore, the well established model of postcrush sciatic nerve regeneration was used to test the hypothesis that LAR is required for neurite outgrowth in the adult mammalian nervous system. In uninjured sciatic nerves, no differences in nerve morphology and sensory function were detected between wild type and LAR-deficient littermate transgenic mice. Sciatic nerve crush resulted in increased LAR protein expression in DRG neurons. In addition, nerve injury led to an increase in the proportion of LAR protein isoforms known to have increased binding affinity to neurite-promoting laminin-nidogen complexes. Two weeks after nerve crush, morphological analysis of distal nerve segments in LAR-deficient transgenic mice demonstrated significantly decreased densities of myelinated fibers, decreased axonal areas, and increased myelin/axon area ratios compared with littermate controls. Electron microscopy analysis revealed a significant twofold reduction in the density of regenerating unmyelinated fibers in LAR-/- nerves distal to the crush site. Sensory testing at the 2 week time point revealed a corresponding 3 mm lag in the proximal-to-distal progression of functioning sensory fibers along the distal nerve segment. These studies introduce PTP receptors as a major new gene family regulating regenerative neurite outgrowth in vivo in the adult mammalian system. PMID- 11438589 TI - Abnormal development of dendritic spines in FMR1 knock-out mice. AB - Fragile X syndrome is caused by a mutation in the FMR1 gene leading to absence of the fragile X mental retardation protein (FMRP). Reports that patients and adult FMR1 knock-out mice have abnormally long dendritic spines of increased density suggested that the disorder might involve abnormal spine development. Because spine length, density, and motility change dramatically in the first postnatal weeks, we analyzed these properties in mutant mice and littermate controls at 1, 2, and 4 weeks of age. To label neurons, a viral vector carrying the enhanced green fluorescent protein gene was injected into the barrel cortex. Layer V neurons were imaged on a two-photon laser scanning microscope in fixed tissue sections. Analysis of >16,000 spines showed clear developmental patterns. Between 1 and 4 weeks of age, spine density increased 2.5-fold, and mean spine length decreased by 17% in normal animals. Early during cortical synaptogenesis, pyramidal cells in mutant mice had longer spines than controls. At 1 week, spine length was 28% greater in mutants than in controls. At 2 weeks, this difference was 10%, and at 4 weeks only 3%. Similarly, spine density was 33% greater in mutants than in controls at 1 week of age. At 2 or 4 weeks of age, differences were not detectable. The spine abnormality was not detected in neocortical organotypic cultures. The transient nature of the spine abnormality in the intact animal suggests that FMRP might play a role in the normal process of dendritic spine growth in coordination with the experience-dependent development of cortical circuits. PMID- 11438590 TI - The role of dopamine receptors in regulating the size of axonal arbors. AB - Factors that regulate terminal arbor size of substantia nigra pars compacta (SNpc) neurons during development and after injury are not well understood. This study examined the role of dopamine receptors in regulating arbor size. Terminal arbors were examined in mice with targeted deletion of the D1 or D2 dopamine receptor [D1(-/-) and D2(-/-) mice, respectively]. Terminal trees were also examined after treatment with receptor blockers and after partial SNpc lesions. Immunohistochemistry was performed, and the number of SNpc neurons and dopaminergic terminals in the striatum was estimated. The number of dopaminergic SNpc neurons were reduced in D1(-/-) and D2(-/-) mice. Density of dopaminergic terminals was unchanged in D1(-/-) mice and increased in D2 (-/-) mice. Steady state striatal DA and DOPAC levels revealed that dopamine activity was enhanced in D2(-/-) mice but reduced in D1(-/-) mice. Two months after partial SNpc lesions, striatal terminal density was normal in both wild-type and D1(-/-) mice but reduced in D2(-/-) mice. Administration of DA receptor antagonists resulted in larger terminal arbors in D1(-/-) and wild-type mice, whereas D2(-/-) mice showed no change in terminal density. Functional blockade of the D2R during development or in the adult brain results in increased axonal sprouting. Partial SNpc lesions resulted in compensatory sprouting, only in mice with functional D2R. These results suggest that individual dopaminergic axons in D2(-/-) mice have reached maximal arbor size. We conclude that the D2 receptor may play a role in modulating the extent of the terminal arbor of SNpc neurons. PMID- 11438591 TI - A critical role of the strychnine-sensitive glycinergic system in spontaneous retinal waves of the developing rabbit. AB - In the developing vertebrate retina, spontaneous electric activity occurs rhythmically in the form of propagating waves and is believed to play a critical role in activity-dependent visual system development, including the establishment of precise retinal and geniculate circuitry. To elucidate how spontaneous retinal waves encode specific developmental cues at various developmental stages, it is necessary to understand how the waves are generated and regulated. Using Ca(2+) imaging and patch clamp in a flat-mount perinatal rabbit retinal preparation, this study demonstrates that, in addition to the cholinergic system, a strychnine sensitive system in the inner retina plays an obligatory and developmentally regulated role in the initiation and propagation of spontaneous retinal waves. This system, which is believed to be the glycinergic network, provided an excitatory drive during early retinal development. It then became inhibitory after postnatal day 1 (P1) to P2, an age when a number of coordinated transitions in neurotransmitter systems occurred concomitantly, and finally contributed to the complete inhibition and disappearance of spontaneous waves after P7-P9. This glycinergic contribution was notably distinct from that of the ionotropic GABAergic system, which was found to exert an inhibitory but nonessential influence on the early wave formation. Blocking glycine- and GABA-gated anion currents had opposing effects on spontaneous retinal waves between embryonic day 29 and P0, suggesting that Cl(-) transporters, particularly R(+)-butylindazone sensitive K-Cl cotransporters, may have a synapse- and/or cell type-specific distribution pattern, in addition to an age-dependent expression pattern in the inner retina. Overall, the results revealed an important reliance of spontaneous retinal waves on dynamic and coordinated interactions among multiple, nonredundant neurotransmitter systems. PMID- 11438592 TI - Stages of synapse development defined by dependence on F-actin. AB - It has been widely speculated that actin plays a central role in CNS synapse assembly, but such a requirement for actin filaments (F-actin) has not yet been demonstrated experimentally. We used hippocampal neurons grown in culture and the actin depolymerizing agent, latrunculin A, to examine directly the relationship between F-actin and synapse formation and maturation. During the first week in culture, actin depolymerization results in a near complete loss of synapses defined by synaptophysin-labeled vesicle clusters, synaptic vesicle recycling, and ultrastructure. Over the second week in culture, F-actin becomes increasingly stable, but actin depolymerization no longer disrupts basic synaptic structure. There is, however, a reduction in the number and size of synaptophysin-labeled clusters and in the size of vesicle clusters undergoing FM4-64 recycling, suggesting that synaptic vesicle anchoring remains partially dependent on F actin. By 18 d in culture, synaptophysin clusters and synaptic vesicle recycling are largely resistant to F-actin depolymerization. The decrease in synapse dependence on F-actin correlates well with the acquisition and retention of presynaptic scaffolding proteins such as Bassoon and postsynaptic scaffolding proteins such as those of the postsynaptic density-95 family. Increased activity stabilizes F-actin and its associated proteins at synaptic sites, suggesting a correlation between active synapses, actin stability, and synapse stability. Our findings demonstrate that F-actin is essential for the development and maintenance of young synapses. Because F-actin is also highly regulatable, we propose that F-actin may be a principal target for stabilizing or destabilizing signals that ultimately result in synapse maintenance or elimination. PMID- 11438593 TI - Long-lasting enhancement of glutamatergic synaptic transmission by acetylcholine contrasts with response adaptation after exposure to low-level nicotine. AB - Attempts to mimic synaptic delivery of acetylcholine (ACh) with brief, repetitive pulses of high concentration ACh at synapses of medial habenula (MHN) and interpeduncular nucleus (IPN) neurons in vitro elicited temporally distinct facilitation and inhibition of glutamate secretion via nicotinic and muscarinic ACh receptor-mediated pathways, respectively. ACh-induced nicotinic facilitation was sustained for up to 2 hr, whereas muscarinic inhibition was transient. Prolonged exposure to nicotine inactivated nicotinic receptors selectively, thus decreasing the relative contribution of the facilitatory versus inhibitory influences of ACh. The net effect of ACh in modulating glutamatergic transmission at MHN-IPN synapses may be determined by pre-exposure to nicotine, because the drug appears to switch the balance between the facilitatory and inhibitory actions of ACh. PMID- 11438594 TI - alpha2-chimaerin, a Cdc42/Rac1 regulator, is selectively expressed in the rat embryonic nervous system and is involved in neuritogenesis in N1E-115 neuroblastoma cells. AB - Neuronal differentiation involves Rac and Cdc42 GTPases. alpha-Chimaerin, a Rac/Cdc42 regulator, occurs as alpha1- and alternatively spliced Src homology 2 (SH2) domain-containing alpha2-isoforms. alpha2-chimaerin mRNA was highly expressed in the rat embryonic nervous system, especially in early postmitotic neurons. alpha1-chimaerin mRNA was undetectable before embryonic day 16.5. Adult alpha2-chimaerin mRNA was restricted to neurons within specific brain regions, with highest expression in the entorhinal cortex. alpha2-chimaerin protein localized to neuronal perikarya, dendrites, and axons. The overall pattern of alpha2-chimaerin mRNA expression resembles that of cyclin-dependent kinase regulator p35 (CDK5/p35) which participates in neuronal differentiation and with which chimaerin interacts. To determine whether alpha2-chimaerin may have a role in neuronal differentiation and the relevance of the SH2 domain, the morphological effects of both chimaerin isoforms were investigated in N1E-115 neuroblastoma cells. When plated on poly-lysine, transient alpha2-chimaerin but not alpha1-chimaerin transfectants formed neurites. Permanent alpha2-chimaerin transfectants generated neurites whether or not they were stimulated by serum starvation, and many cells were enlarged. Permanent alpha1-chimaerin transfectants displayed numerous microspikes and contained F-actin clusters, a Cdc42-phenotype, but generated few neurites. In neuroblastoma cells, alpha2 chimaerin was predominantly soluble with some being membrane-associated, whereas alpha1-chimaerin was absent from the cytosol, being membrane- and cytoskeleton associated, paralleling their subcellular distribution in brain. Transient transfection with alpha2-chimaerin mutated in the SH2 domain (N94H) generated an alpha1-chimaerin-like phenotype, protein partitioned in the particulate fraction, and in NGF-stimulated pheochromocytoma cell line 12 (PC12) cells, neurite formation was inhibited. These results indicate a role for alpha2-chimaerin in morphological differentiation for which its SH2 domain is vital. PMID- 11438595 TI - How simple cells are made in a nonlinear network model of the visual cortex. AB - Simple cells in the striate cortex respond to visual stimuli in an approximately linear manner, although the LGN input to the striate cortex, and the cortical network itself, are highly nonlinear. Although simple cells are vital for visual perception, there has been no satisfactory explanation of how they are produced in the cortex. To examine this question, we have developed a large-scale neuronal network model of layer 4Calpha in V1 of the macaque cortex that is based on, and constrained by, realistic cortical anatomy and physiology. This paper has two aims: (1) to show that neurons in the model respond like simple cells. (2) To identify how the model generates this linearized response in a nonlinear network. Each neuron in the model receives nonlinear excitation from the lateral geniculate nucleus (LGN). The cells of the model receive strong (nonlinear) lateral inhibition from other neurons in the model cortex. Mathematical analysis of the dependence of membrane potential on synaptic conductances, and computer simulations, reveal that the nonlinearity of corticocortical inhibition cancels the nonlinear excitatory input from the LGN. This interaction produces linearized responses that agree with both extracellular and intracellular measurements. The model correctly accounts for experimental results about the time course of simple cell responses and also generates testable predictions about variation in linearity with position in the cortex, and the effect on the linearity of signal summation, caused by unbalancing the relative strengths of excitation and inhibition pharmacologically or with extrinsic current. PMID- 11438596 TI - Altered respiratory activity and respiratory regulations in adult monoamine oxidase A-deficient mice. AB - The abnormal metabolism of serotonin during the perinatal period alters respiratory network maturation at birth as revealed by comparing the monoamine oxidase A-deficient transgenic (Tg8) with the control (C3H) mice (Bou-Flores et al., 2000). To know whether these alterations occur only transiently or induce persistent respiratory dysfunction during adulthood, we studied the respiratory activity and regulations in adult C3H and Tg8 mice. First, plethysmographic and pneumotachographic analyses of breathing patterns revealed weaker tidal volumes and shorter inspiratory durations in Tg8 than in C3H mice. Second, electrophysiological studies showed that the firing activity of inspiratory medullary neurons and phrenic motoneurons is higher in Tg8 mice and that of the intercostal motoneurons in C3H mice. Third, histological studies indicated abnormally large cell bodies of Tg8 intercostal but not phrenic motoneurons. Finally, respiratory responses to hypoxia and lung inflation are weaker in Tg8 than in C3H mice. dl-p-chlorophenyl-alanine treatments applied to Tg8 mice depress the high serotonin level present during adulthood; the treated mice recover normal respiratory responses to both hypoxia and lung inflation, but their breathing parameters are not significantly affected. Therefore in Tg8 mice the high serotonin level occurring during the perinatal period alters respiratory network maturation and produces a permanent respiratory dysfunction, whereas the high serotonin level present in adults alters the respiratory regulatory processes. In conclusion, the metabolism of serotonin plays a crucial role in the maturation of the respiratory network and in both the respiratory activity and the respiratory regulations. PMID- 11438597 TI - Amygdala is critical for stress-induced modulation of hippocampal long-term potentiation and learning. AB - Stress is a biologically significant factor shown to influence synaptic plasticity and memory functioning in the hippocampus. This study examined the role of the amygdala, a brain structure implicated in coordinating stress behaviors and modulating memory consolidation, in mediating stress effects on hippocampal long-term potentiation (LTP) and memory in rats. Electrolytic lesions of the amygdala effectively blocked the adverse physiological and behavioral effects of restraint and tailshock stress, without impeding the increase in corticosterone secretion to stress. Physiologically, hippocampal slices from stressed animals exhibited impaired LTP relative to slices from unstressed control animals, whereas hippocampal slices from stressed animals with amygdalar lesions exhibited normal LTP. Behaviorally, stressed animals were impaired in retention of a hippocampal-dependent hidden platform version of the Morris water maze task, and this impairment was blocked by amygdalar lesions. In a fixed location-visible platform water maze task that can be acquired by independent hippocampal and nonhippocampal memory systems, stress enhanced the use of nonhippocampal-based memory to acquire the task. These results indicate that an intact amygdala is necessary for the expression of the modulatory effects of stress on hippocampal LTP and memory. PMID- 11438598 TI - Global structure, robustness, and modulation of neuronal models. AB - The electrical characteristics of many neurons are remarkably robust in the face of changing internal and external conditions. At the same time, neurons can be highly sensitive to neuromodulators. We find correlates of this dual robustness and sensitivity in a global analysis of the structure of a conductance-based model neuron. We vary the maximal conductance parameters of the model neuron and, for each set of parameters tested, characterize the activity pattern generated by the cell as silent, tonically firing, or bursting. Within the parameter space of the five maximal conductances of the model, we find directions, representing concerted changes in multiple conductances, along which the basic pattern of neural activity does not change. In other directions, relatively small concurrent changes in a few conductances can induce transitions between these activity patterns. The global structure of the conductance-space maps implies that neuromodulators that alter a sensitive set of conductances will have powerful, and possibly state-dependent, effects. Other modulators that may have no direct impact on the activity of the neuron may nevertheless change the effects of such direct modulators via this state dependence. Some of the results and predictions arising from the model studies are replicated and verified in recordings of stomatogastric ganglion neurons using the dynamic clamp. PMID- 11438599 TI - Hyperactivity and intact hippocampus-dependent learning in mice lacking the M1 muscarinic acetylcholine receptor. AB - Members of the muscarinic acetylcholine receptor family are thought to play key roles in the regulation of a large number of important functions of the CNS. However, the precise roles of the individual muscarinic receptor subtypes in modulating these processes are not well understood at present, primarily because of the lack of ligands with sufficient receptor subtype selectivity. To investigate the behavioral significance of the M(1) muscarinic receptor (M(1)R), which is abundantly expressed in the forebrain, we subjected M(1) receptor deficient mice (M(1)R(-/-) mice) to a battery of behavioral tests. M(1)R(-/-) mice showed no significant impairments in neurological reflexes, motor coordination, pain sensitivity, and prepulse inhibition. Strikingly, however, M(1)R(-/-) mice consistently exhibited a pronounced increase in locomotor activity in various tests, including open field, elevated plus maze, and light/dark transition tests. Moreover, M(1)R(-/-) mice showed reduced immobilization in the Porsolt forced swim test and reduced levels of freezing after inescapable footshocks, suggesting that M(1)R(-/-) mice are hyperactive under stressful conditions as well. An increased number of social contacts was observed in a social interaction test. Surprisingly, M(1)R(-/-) mice displayed no significant cognitive impairments in the Morris water maze and in contextual fear conditioning. M(1)R(-/-) mice showed slight performance deficits in auditory-cued fear conditioning and in an eight-arm radial maze, most likely because of the hyperactivity phenotype displayed by the M(1)R(-/-) mice. Our results indicate that M(1) muscarinic receptors play an important role in the regulation of locomotor activity but appear to be less critical for cognitive processes, as generally assumed. PMID- 11438600 TI - Bilateral integration of whisker information in the primary somatosensory cortex of rats. AB - The isomorphic representation of the contralateral whisker pad in the rodent cerebral cortex has served as a canonical example in primary somatosensory areas that the contralateral body surface is spatially represented as a topographic map. By characterizing responses evoked by multiwhisker stimuli, we provide direct evidence that the whisker region of the rat primary somatosensory cortex (SI) integrates information from both contralateral and ipsilateral whisker pads. The proportions of SI neurons responsive to ipsilateral whisker stimuli, as well as their response probabilities, increased with the number of ipsilateral whiskers stimulated. Under bilateral whisker stimulation, the responses of 95% of neurons recorded were affected by stimulation of ipsilateral whiskers. Contralateral tactile responses of SI neurons were profoundly influenced by preceding ipsilateral stimuli and vice versa. This effect depended on both the spatial location and the relative timing of bilateral whisker stimuli, leading to both spatial and temporal asymmetries of interaction. Bilateral whisker stimulation resulted in only modest changes in evoked response latency. Previous ipsilateral stimulation was also shown to affect tactile responses evoked by later ipsilateral stimuli. Inactivation of the opposite SI abolished ipsilaterally evoked responses as well as their influence on subsequently evoked contralateral responses in the intact SI. Based on these results, we conclude that the rat SI integrates information from both whisker pads and propose that such interactions may underlie the ability of rats to discriminate bilateral tactile stimuli. PMID- 11438601 TI - Attention systems and the organization of the human parietal cortex. AB - Event-related functional magnetic resonance imaging was used to compare activity in the human parietal cortex in two attention-switching paradigms. On each trial of the visual switching (VS) paradigm, subjects attended to one of two visual stimuli on the basis of either their color or shape. Trials were presented in blocks interleaved with cues instructing subjects to either continue attending to the currently relevant dimension or to switch to the other stimulus dimension. In the response switching (RS) paradigm, subjects made one of two manual responses to the single stimulus presented on each trial. The rules for stimulus-response mapping were reversed on different trials. Trials were presented in blocks interleaved with cues that instructed subjects to either switch stimulus-response mapping rules or to continue with the current rule. Brain activity at "switch" and "stay" events was compared. The results revealed distinct parietal areas concerned with visual attentional set shifts (VS) and visuomotor intentional set shifts (RS). In VS, activity was recorded in the lateral part of the intraparietal region. In RS, activity was recorded in the posterior medial intraparietal region and adjacent posterior superior and dorsomedial parietal cortex. The results also suggest that the basic functional organization of the intraparietal sulcus and surrounding regions is similar in both macaque and human species. PMID- 11438602 TI - Enriched rehabilitative training promotes improved forelimb motor function and enhanced dendritic growth after focal ischemic injury. AB - Chronic impairment of forelimb and digit movement is a common problem after stroke that is resistant to therapy. Previous studies have demonstrated that enrichment improves behavioral outcome after focal ischemia; however, postischemic enrichment alone is not capable of enhancing fine digit and forelimb function. Therefore, we combined environmental enrichment with daily skilled reach training to assess the effect of intensive task-specific rehabilitation on long-term functional outcome. Rats were subjected to either endothelin-1-induced focal ischemia or sham surgery and subsequently designated to enriched rehabilitation or standard-housing treatment groups starting 15 d after ischemia. Functional assessment of the affected forelimb at 4 and 9 weeks after treatment revealed that ischemic plus enrichment (IE) animals had improved approximately 30% on the staircase-reaching task and were indistinguishable from sham animals for both latency and foot faults in a beam-traversing task. In contrast, ischemic plus standard (IS) animals remained significantly impaired on both tasks. Interestingly, both ischemic groups (IE and IS) relied on the nonaffected forelimb during upright weight-bearing movements, a pattern that persisted for the duration of the experiment. Dendritic arborization of layer V pyramidal cells within the undamaged motor cortex was examined using a Golgi-Cox procedure. IE animals showed enhanced dendritic complexity and length compared with both IS and sham groups. These results suggest that enrichment combined with task-specific rehabilitative therapy is capable of augmenting intrinsic neuronal plasticity within noninjured, functionally connected brain regions, as well as promoting enhanced functional outcome. PMID- 11438603 TI - Inhibition of neuropathic pain by selective ablation of brainstem medullary cells expressing the mu-opioid receptor. AB - Neurons in the rostroventromedial medulla (RVM) project to spinal loci where the neurons inhibit or facilitate pain transmission. Abnormal activity of facilitatory processes may thus represent a mechanism of chronic pain. This possibility and the phenotype of RVM cells that might underlie experimental neuropathic pain were investigated. Cells expressing mu-opioid receptors were targeted with a single microinjection of saporin conjugated to the mu-opioid agonist dermorphin; unconjugated saporin and dermorphin were used as controls. RVM dermorphin-saporin, but not dermorphin or saporin, significantly decreased cells expressing mu-opioid receptor transcript. RVM dermorphin, saporin, or dermorphin-saporin did not change baseline hindpaw sensitivity to non-noxious or noxious stimuli. Spinal nerve ligation (SNL) injury in rats pretreated with RVM dermorphin-saporin failed to elicit the expected increase in sensitivity to non noxious mechanical or noxious thermal stimuli applied to the paw. RVM dermorphin or saporin did not alter SNL-induced experimental pain, and no pretreatment affected the responses of sham-operated groups. This protective effect of dermorphin-saporin against SNL-induced pain was blocked by beta-funaltrexamine, a selective mu-opioid receptor antagonist, indicating specific interaction of dermorphin-saporin with the mu-opioid receptor. RVM microinjection of dermorphin saporin, but not of dermorphin or saporin, in animals previously undergoing SNL showed a time-related reversal of the SNL-induced experimental pain to preinjury baseline levels. Thus, loss of RVM mu receptor-expressing cells both prevents and reverses experimental neuropathic pain. The data support the hypothesis that inappropriate tonic-descending facilitation may underlie some chronic pain states and offer new possibilities for the design of therapeutic strategies. PMID- 11438604 TI - Temporal cues contribute to tactile perception of roughness. AB - Optimal perception of surface roughness requires lateral movement between skin and surface, suggesting the importance of temporal cues. The roughness of periodic gratings is affected by changing either inter-element spacing (groove width, G) or element width (ridge width, R). Peripheral neural responses to gratings depend quantitatively on a spatial variable, G, and a temporal variable, grating temporal frequency (F(t)), with changes in R acting indirectly through concomitant changes in F(t). We investigated, psychophysically, the contribution of temporal cues to human tactile perception of roughness, using gratings varying in either R or G. Gratings were scanned across the immobile fingerpad with controlled movement speed (S) and contact force. In one experiment, we found that roughness magnitude estimates depended on both G and F(t). In a second experiment, discrimination of the roughness of gratings varying in either R or G was affected by manipulating F(t). Overall, the effect of G on roughness judgments was much stronger than that of F(t), probably explaining why many previous studies using surfaces that varied only in inter-element spacing led to the conclusion that temporal factors play no role in roughness perception. However, the perceived roughness of R-varying gratings was determined by F(t) and not spatial variables. Roughness judgments were influenced by G and F(t) in a manner entirely consistent with predicted afferent response rates. Thus perceived roughness, like peripheral afferent responses, depends in part on temporal variables. PMID- 11438605 TI - The cAMP-protein kinase A signal transduction pathway modulates ethanol consumption and sedative effects of ethanol. AB - Ethanol and other drugs of abuse modulate cAMP-PKA signaling within the mesolimbic reward pathway. To understand the role of the cAMP-PKA signal transduction in mediating the effects of ethanol, we have studied ethanol consumption and the sedative effects of ethanol in three lines of genetically modified mice. We report that mice with the targeted disruption of one Gsalpha allele as well as mice with reduced neuronal PKA activity have decreased alcohol consumption compared with their wild-type littermates. Genetic reduction of cAMP PKA signaling also makes mice more sensitive to the sedative effects of ethanol, although plasma ethanol concentrations are unaffected. In contrast, mice with increased adenylyl cyclase activity resulting from the transgenic expression of a constitutively active form of Gsalpha in neurons within the forebrain are less sensitive to the sedative effects of ethanol. Thus, the cAMP-PKA signal transduction pathway is critical in modulating sensitivity to the sedative effects of ethanol as well as influencing alcohol consumption. PMID- 11438606 TI - Involvement of human amygdala and orbitofrontal cortex in hunger-enhanced memory for food stimuli. AB - We used positron emission tomography to measure regional cerebral blood flow (rCBF) in 10 healthy volunteers performing a recognition memory task with food and non-food items. The biological salience of the food stimuli was manipulated by requiring subjects to fast before the experiment and eat to satiation at fixed time points during scanning. All subjects showed enhanced recognition of food stimuli (relative to non-food) in the fasting state. Satiation significantly reduced the memory advantage for food. Left amygdala rCBF covaried positively with recognition memory for food items, whereas rCBF in right anterior orbitofrontal cortex covaried with overall memory performance. Right posterior orbitofrontal rCBF covaried positively with hunger ratings during presentation of food items. Regression analysis of the neuroimaging data revealed that left amygdala and right lateral orbitofrontal rCBF covaried as a function of stimulus category (i.e., food vs non-food). These results indicate the involvement of amygdala and discrete regions of orbitofrontal cortex in the integration of perceptual (food), motivational (hunger), and cognitive (memory) processes in the human brain. PMID- 11438607 TI - Membrane bistability in olfactory bulb mitral cells. AB - Whole-cell patch-clamp recordings were used to investigate the electrophysiological properties of mitral cells in rat main olfactory bulb brain slice preparations. The majority of mitral cells are bistable. These cells spontaneously alternate between two membrane potentials, separated by approximately 10 mV: a relatively depolarized potential (upstate), which is perithreshold for spike generation, and a relatively hyperpolarized potential (downstate), in which spikes do not occur. Bistability occurs spontaneously in the absence of ionotropic excitatory or inhibitory synaptic inputs. Bistability is voltage dependent; transition from the downstate to the upstate is a regenerative event activated by brief depolarization. A brief hyperpolarization can switch the membrane potential from the upstate to the downstate. In response to olfactory nerve (ON) stimulation, mitral cells in the upstate are more likely to fire an action potential than are those in the downstate. ON stimulation can switch the membrane potential from the downstate to the upstate, producing a prolonged and amplified depolarization in response to a brief synaptic input. We conclude that bistability is an intrinsic property of mitral cells that is a major determinant of their responses to ON input. PMID- 11438608 TI - PKCgamma contributes to a subset of the NMDA-dependent spinal circuits that underlie injury-induced persistent pain. AB - In previous studies we provided evidence that the gamma isoform of protein kinase C (PKCgamma) is an important contributor to the increased pain sensitivity that occurs after injury. Here we combined electrophysiological and behavioral approaches in wild-type and PKCgamma-null mice to compare the hyperexcitability of wide dynamic range neurons in lamina V of the spinal cord dorsal horn with the behavioral hyperexcitability produced by the same injury [application of a C fiber irritant, mustard oil (MO), to the hindpaw]. Wild-type and null mice did not differ in their response to mechanical or thermal stimuli before tissue injury, and the magnitude of the response to the MO stimuli was comparable. In wild-type mice, MO produced a dramatic and progressive enhancement of the response of lamina V neurons to innocuous mechanical and thermal stimuli. The time course of the neuronal hyperexcitability paralleled the time course of the MO-induced behavioral allodynia (nocifensive behavior in response to a previously innocuous mechanical stimulus). Neuronal hyperexcitability was also manifest in the PKCgamma-null mice, but it lasted <30 min. By contrast, the behavioral allodynia produced by MO in the PKCgamma-null mice, although reduced to approximately half that of the wild-type mice, persisted long after the lamina V hyperexcitability had subsided. Because the MO-induced behavioral allodynia was completely blocked by an NMDA receptor antagonist, we conclude that PKCgamma mediates the transition from short- to long-term hyperexcitability of lamina V nociresponsive neurons but that the persistence of injury-induced pain must involve activity within multiple NMDA-dependent spinal cord circuits. PMID- 11438609 TI - Negative interspike interval correlations increase the neuronal capacity for encoding time-dependent stimuli. AB - Accurate detection of sensory input is essential for the survival of a species. Weakly electric fish use amplitude modulations of their self-generated electric field to probe their environment. P-type electroreceptors convert these modulations into trains of action potentials. Cumulative relative refractoriness in these afferents leads to negatively correlated successive interspike intervals (ISIs). We use simple and accurate models of P-unit firing to show that these refractory effects lead to a substantial increase in the animal's ability to detect sensory stimuli. This assessment is based on two approaches, signal detection theory and information theory. The former is appropriate for low frequency stimuli, and the latter for high-frequency stimuli. For low frequencies, we find that signal detection is dependent on differences in mean firing rate and is optimal for a counting time at which spike train variability is minimal. Furthermore, we demonstrate that this minimum arises from the presence of negative ISI correlations at short lags and of positive ISI correlations that extend out to long lags. Although ISI correlations might be expected to reduce information transfer, in fact we find that they improve information transmission about time-varying stimuli. This is attributable to the differential effect that these correlations have on the noise and baseline entropies. Furthermore, the gain in information transmission rate attributable to correlations exhibits a resonance as a function of stimulus bandwidth; the maximum occurs when the inverse of the cutoff frequency of the stimulus is of the order of the decay time constant of refractory effects. Finally, we show that the loss of potential information caused by a decrease in spike-timing resolution is smaller for low stimulus cutoff frequencies than for high ones. This suggests that a rate code is used for the encoding of low-frequency stimuli, whereas spike timing is important for the encoding of high-frequency stimuli. PMID- 11438610 TI - Functional interaction between opioid and cannabinoid receptors in drug self administration. AB - The present study was designed to explore the relationship between the cannabinoid and opioid receptors in animal models of opioid-induced reinforcement. The acute administration of SR141716A, a selective central cannabinoid CB1 receptor antagonist, blocked heroin self-administration in rats, as well as morphine-induced place preference and morphine self-administration in mice. Morphine-dependent animals injected with SR141716A exhibited a partial opiate-like withdrawal syndrome that had limited consequences on operant responses for food and induced place aversion. These effects were associated with morphine-induced changes in the expression of CB1 receptor mRNA in specific nuclei of the reward circuit, including dorsal caudate putamen, nucleus accumbens, and septum. Additionally, the opioid antagonist naloxone precipitated a mild cannabinoid-like withdrawal syndrome in cannabinoid-dependent rats and blocked cannabinoid self-administration in mice. Neither SR141716A nor naloxone produced any intrinsic effect on these behavioral models. The present results show the existence of a cross-interaction between opioid and cannabinoid systems in behavioral responses related to addiction and open new strategies for the treatment of opiate dependence. PMID- 11438611 TI - In vivo resetting of the hamster circadian clock by 5-HT7 receptors in the suprachiasmatic nucleus. AB - Serotonin (5-HT) has been strongly implicated in the regulation of the mammalian circadian clock located in the suprachiasmatic nuclei (SCN); however, its role in behavioral (nonphotic) circadian phase resetting remains elusive. Central to this issue are divergent lines of evidence that the SCN may, or may not, be a target for the phase-resetting effects of 5-HT. We have addressed this question using a novel reverse-microdialysis approach for timed perfusions of serotonergic and other agents to the Syrian hamster SCN with durations equivalent to the increases in in vivo 5-HT release during phase-resetting behavioral manipulations. We found that 3 hr perfusions of the SCN with either 5-HT or the 5-HT(1A,7) receptor agonist 2-dipropylamino-8-hydroxy-1,2,3,4-tetrahydro-naphthalene (8-OH-DPAT) at midday advanced the phase of the free-running circadian rhythm of wheel-running assessed using an Aschoff type II procedure. Phase shifts induced by 8-OH-DPAT were enhanced more than threefold by pretreatment with the 5-HT synthesis inhibitor para-chlorophenylalanine. Phase advances induced by SCN 8-OH-DPAT perfusion were significantly inhibited by the 5-HT(2,7) receptor antagonist ritanserin and by the more selective 5-HT(7) receptor antagonist DR4004, implicating the 5-HT(7) receptor in mediating this phase resetting. Concurrent exposure to light during the 8-OH-DPAT perfusion abolished the phase advances. Furthermore, coperfusion of the SCN with TTX, which blocked in vivo 5-HT release, did not suppress intra-SCN 8-OH-DPAT-induced phase advances. These results indicate that 5-HT(7) receptor-mediated phase resetting in the SCN is markedly influenced by the degree of postsynaptic responsiveness to 5-HT and by photic stimulation. Finally, 5-HT may act directly on SCN clock cells to induce in vivo nonphotic phase resetting. PMID- 11438612 TI - Local injection of endothelin-1 produces pain-like behavior and excitation of nociceptors in rats. AB - Neurobehavioral and neurophysiological actions of the peptide endothelin-1 (ET-1) were investigated after subcutaneous plantar hindpaw injections in adult male Sprague Dawley rats. Hindpaw flinching developed within minutes after ET-1 (8-16 nmol) injection, peaked at 30 min, lasted for 60 min, and was strongly inhibited by the endothelin-A (ET(A)) receptor antagonist, BQ-123 (3.2 m). In separate experiments, impulse activity of single, physiologically characterized sensory C , Adelta-, and Abeta-fibers was recorded from the sciatic nerve in anesthetized rats after subcutaneous injections of endothelin-1 (1-20 nmol), alone or together with BQ-123 (3.2 m), into the plantar hindpaw receptive fields of these units. All nociceptive C-fibers (31 of 33 C-fibers studied) were excited by ET-1 (1-20 nmol) in a dose-dependent manner. For doses of 16-20 nmol, the mean latency for afferent activation after injection of ET-1 was 3.16 +/- 0.31 min, and the mean and maximum response frequency were 2.02 +/- 0.48 impulses (imp)/sec and 14.0 +/- 3.2 imp/sec, respectively. All 10 nociceptive Adelta-fibers (of 12 Adelta-fibers studied) also responded to 1-20 nmol of ET-1 in a dose-dependent manner with a mean latency of 3.5 +/- 0.12 min and mean response frequency of 3.3 +/- 2.3 imp/sec. In contrast, most Abeta-fibers (9 of 12) did not respond to ET-1. BQ 123, when coinjected with ET-1, blocked ET-1-induced activation in all C- and Adelta-fibers tested. These data demonstrate that subcutaneous administration of ET-1 to the rat plantar hindpaw produces pain-like behavior and selective excitation of nociceptive fibers through activation of ET(A) receptors. PMID- 11438613 TI - The neuropeptide Y Y5 receptor mediates the blockade of "photic-like" NMDA induced phase shifts in the golden hamster. AB - Circadian or daily rhythms generated from the mammalian suprachiasmatic nuclei (SCN) of the hypothalamus can be synchronized by light and nonphotic stimuli. Whereas glutamate mediates photic information, nonphotic information can in some cases be mediated by neuropeptide Y (NPY) or serotonin. NPY or serotonin can reduce the phase-resetting effect of light or glutamate; however, the mechanisms and level of interaction of these two kinds of stimuli are unknown. Here we investigate the effect of NPY on the NMDA-induced phase shift of the hamster SCN circadian neural activity rhythm by means of single-unit recording techniques. NMDA (10-100 microm) applied in the early subjective night induced phase delays in the time of peak firing, whereas doses in the millimolar range disrupted firing patterns. The NMDA-induced phase delay was blocked by coapplication of NPY (0.02-200 microm). NPY Y1/Y5 and Y5 receptor agonists, but not the Y2 receptor agonist, blocked the NMDA-induced phase delay in a similar manner as NPY. The coapplication of a Y5 but not Y1 receptor antagonist eliminated NPY blockade of NMDA-induced phase delays, suggesting that the Y5 receptor is capable of mediating the inhibitory effect of NPY on photic responses. These results indicate that nonphotic and photic stimuli may interact at a level at or beyond NMDA receptor response and indicate that the Y5 receptor is involved in this interaction. Alteration of Y5 receptor function may therefore be expected to alter synchronization of circadian rhythms to light. PMID- 11438614 TI - Whisker deafferentation and rodent whisking patterns: behavioral evidence for a central pattern generator. AB - Even in the absence of explicit stimulation, rats emit patterns of rhythmic whisking movements. Because of their stereotyped nature and their persistence after sensory denervation and cortical ablation, whisking movements have been assumed to reflect the output of a central pattern generator (CPG). However, identification of a movement pattern as the product of a CPG requires evidence that its generation, patterning, and coordination are independent of sensory input. To provide such evidence, we used optoelectronic instrumentation to obtain high-resolution records of the movement trajectories of individual whiskers in rats whose heads were fixed to isolate their exploratory whisking from exafferent inputs. Unconditioned whisking patterns were quantitatively characterized by a biometric analysis of the kinematics, rhythmicity, and coordination of bilaterally homologous vibrissa movements. Unilateral and bilateral sectioning of the infraorbital nerve, which innervates the whiskers, was then performed to block reafferent inputs generated by the animal's own whisking movements. Unilateral sectioning of the nerve has no effect on whisking kinematics but is followed by a significant but relatively transient bilateral increase in whisking frequency. However, bilateral deafferentation, when performed in a single-stage procedure, does not disrupt the generation, patterning, or bilateral coordination of whisking patterns in the rat. These findings provide strong behavioral evidence for a whisking CPG and are discussed in relation to its possible location and properties. PMID- 11438615 TI - Chronic intermittent hypoxia elicits serotonin-dependent plasticity in the central neural control of breathing. AB - We tested the hypothesis that chronic intermittent hypoxia (CIH) elicits plasticity in the central neural control of breathing via serotonin-dependent effects on the integration of carotid chemoafferent inputs. Adult rats were exposed to 1 week of nocturnal CIH (11-12% O(2)/air at 5 min intervals; 12 hr/night). CIH and untreated rats were then anesthetized, paralyzed, vagotomized, and artificially ventilated. Time-dependent hypoxic responses were assessed in the phrenic neurogram during and after three 5 min episodes of isocapnic hypoxia. Integrated phrenic amplitude (integralPhr) responses during hypoxia were greater after CIH at arterial oxygen pressures (PaO(2)) between 25 and 45 mmHg (p < 0.05), but not at higher PaO(2) levels. CIH did not affect hypoxic phrenic burst frequency responses, although the post-hypoxia frequency decline that is typical in rats was abolished. integralPhr and frequency responses to electrical stimulation of the carotid sinus nerve were enhanced by CIH (p < 0.05). Serotonin dependent long-term facilitation (LTF) of integralPhr was enhanced after CIH at 15, 30, and 60 min after episodic hypoxia (p < 0.05). Pretreatment with the serotonin receptor antagonists methysergide (4 mg/kg, i.v.) and ketanserin (2 mg/kg, i.v.) reversed CIH-induced augmentation of the short-term hypoxic phrenic response and restored the post-hypoxia frequency decline in CIH rats. Whereas methysergide abolished CIH-enhanced phrenic LTF, the selective 5-HT(2) antagonist ketanserin only partially reversed this effect. The results suggest that CIH elicits unique forms of serotonin-dependent plasticity in the central neural control of breathing. Enhanced LTF after CIH may involve an upregulation of a non 5-HT(2) serotonin receptor subtype or subtypes. PMID- 11438616 TI - 18F-FDG PET detection of colonic adenomas. AB - The adenomatous polyp of the colon is clinically important as a precursor of colonic cancer. The aim of this preliminary study was to evaluate the potential usefulness of (18)F-FDG PET for detecting adenomatous polyps of the colon. METHODS: We performed a retrospective study of 110 subjects who underwent both PET study and total colonoscopy. On nonattenuation-corrected PET images, focal distinct FDG accumulation along the large intestine was considered a positive finding, and the PET results were compared with colonoscopic findings. Histology and adenoma size were determined by polypectomy. RESULTS: Fifty-nine adenomatous polyps, 5-30 mm in size, were found in 30 subjects by total colonoscopy. PET findings were positive for 14 of the 59 adenomas (24%). The positivity rate for PET images rose with the increase in size of the adenomas; it was 90% in adenomas (9/10) that were > or =13 mm. The overall false-positive rate was 5.5% (6/110 subjects). CONCLUSION: Increased glucose metabolism is observed in colonic adenomas, and detectability with PET increases with the increase in adenoma size. Adenomas are premalignant lesions, and it is important to realize that colonic adenomas may be found incidentally during an FDG PET study. PMID- 11438617 TI - Chromogranin A assay and (131)I-MIBG scintigraphy for diagnosis and follow-up of pheochromocytoma. AB - We assessed the performance of a new serum chromogranin A (CgA) assay in combination with the results of (131)I-metaiodobenzylguanidine (MIBG) scintigraphy for diagnosis and follow-up in 89 patients with clinical findings suggestive of pheochromocytoma. METHODS: The study population consisted of 41 patients with proven pheochromocytoma and 48 patients with refuted pheochromocytoma. Eighty-seven scintigraphy examinations were performed, 52 in patients with proven pheochromocytoma (39 before surgery and 13 after surgery) and 35 in patients with refuted pheochromocytoma. RESULTS: The sensitivity of the CgA level was 90.2%, and the specificity was 99.0% and 92.3% in the control and refuted pheochromocytoma groups, respectively. A significant relationship was seen between serum levels of CgA and tumor mass (r = 0.70; P < 10(-5)). The postoperative CgA level was an early and accurate predictor of curative surgery or relapse. The concordance between CgA levels and scintigraphic data was 90.8%. CONCLUSION: Serum CgA level is an effective marker of pheochromocytoma. Increased levels strongly correlate with tumor mass; therefore, small tumors may go undetected. The concordance between CgA level and the results of (131)I-MIBG scintigraphy is high. A CgA level in the reference range is highly predictive of normal scintigraphy findings. PMID- 11438618 TI - The new technology of combined transmission and emission tomography in evaluation of endocrine neoplasms. AB - The clinical value of a novel technology of combined transmission and emission tomography (TET) was assessed in patients with endocrine tumors. METHODS: TET technology, which combines simultaneous acquisition of SPECT and CT images, using the same imaging device, allows correct fusion of images of both modalities. TET was performed on 27 patients with known or suspected endocrine tumors. The radiopharmaceuticals used for the emission part of the study were chosen according to the tumor type: (111)In-octreotide for patients with neuroendocrine tumors (n = 10), (99m)Tc-sestamibi for patients with primary hyperparathyroidism (n = 8), (131)I for patients with thyroid cancer (n = 4), and (123)I metaiodobenzylguanidine and (75)Se-cholesterol for patients with adrenal masses (n = 3 and n = 2, respectively). The additional information provided by TET compared with scintigraphy was assessed for both image interpretation and clinical utility. RESULTS: TET did not provide any additional data in 16 patients (59%), including 5 patients with normal scintigraphy. In 11 patients (41%) with abnormal SPECT findings, TET improved image interpretation by providing a better anatomic localization of SPECT-detected lesions. It showed unsuspected bone involvement in 4 patients, it identified the organs involved and the relationship of the lesions to neighboring structures in 5 patients, and it differentiated physiologic uptake from tumor uptake in 2 patients. TET provided additional information of clinical value in 9 patients (33%). It assisted in better planning of surgery in 2 patients with neuroendocrine tumors and in 2 patients with ectopic parathyroid adenomas. It changed the treatment approach in 2 patients with neuroendocrine tumors and 1 patient with thyroid carcinoma, and it altered prognosis in 2 patients with thyroid malignancy. CONCLUSION: TET enhances the already unique role of nuclear medicine procedures in the assessment and management of patients with endocrine neoplasms. PMID- 11438619 TI - Troglitazone improves whole-body insulin resistance and skeletal muscle glucose use in type II diabetic patients. AB - Recently, troglitazone has emerged as an insulin sensitizer for the treatment of type II diabetes. However, its effect on skeletal muscle glucose use (SMGU) has not been studied. METHODS: To investigate the effect of troglitazone on SMGU in patients with type II diabetes, we undertook skeletal muscle (18)F-FDG PET dynamic imaging under insulin clamping before and after administration of SMGU to 20 patients with type II diabetes. Data were compared with those for 12 age matched healthy volunteers. RESULTS: The whole-body glucose disposal rate (GDR) was significantly lower in patients (29.9 +/- 9.83 micromol/min/kg) than in control subjects (55.6 +/- 16.5 micromol/min/kg, P < 0.01), as was the SMGU (patients, 3.27 +/- 2.17 micromol/min/kg; control subjects, 10.9 +/- 6.4 micromol/min/kg; P < 0.01). After the therapy, GDR significantly improved in patients (29.3 +/- 14.6 micromol/min/kg, P < 0.05), as did SMGU (5.06 +/- 2.11 micromol/min/kg, P < 0.05). When results for patients with and without hypertension were separately analyzed, a significant improvement in SMGU after troglitazone was seen in both normotensive and hypertensive patients (normotensive [n = 10]: baseline, 3.67 +/- 2.89 micromol/min/kg; after therapy, 5.28 +/- 2.61 micromol/min/kg; P < 0.05; hypertensive [n = 10]: baseline, 2.89 +/ 1.22 micromol/min/kg; after therapy, 4.72 +/- 1.39 micromol/min/kg; P < 0.05). GDR in patients with and without hypertension was significantly improved by troglitazone (normotensive: baseline, 17.9 +/- 10.2 micromol/min/kg; after therapy, 31.9 +/- 15.9 micromol/min/kg; P < 0.01; hypertensive: baseline, 39.6 +/ 15.1 micromol/min/kg; after therapy, 47.7 +/- 23.8 micromol/min/kg; P < 0.05). The plasma free fatty acid concentration during insulin clamping was not changed by troglitazone (baseline, 1.1 +/- 0.86 mEq/L; after therapy, 0.93 +/- 0.65 mEq/L; P = not significant). CONCLUSION: Troglitazone can improve whole-body insulin resistance through the improvement of SMGU but not through a decline in plasma free fatty acid concentration in patients with type II diabetes with or without hypertension. PMID- 11438620 TI - Reappearance of cardiac presynaptic sympathetic nerve terminals in the transplanted heart: correlation between PET using (11)C-hydroxyephedrine and invasively measured norepinephrine release. AB - Previously, sympathetic reinnervation of the transplanted heart has been described using invasive catheterization techniques and noninvasive radionuclide imaging techniques. However, little is known about the agreement between these 2 methods. Thus, correlation between (11)C-hydroxyephedrine (HED) PET and invasively measured norepinephrine (NE) release was investigated in transplant recipients in this study. METHODS: Using PET and the catecholamine analog HED, 17 patients were studied between 2 mo and 13.6 y after transplantation. Based on results in completely denervated hearts, areas with HED retention >7%/min were defined as reinnervated. Additionally, transcardiac NE release induced by intravenous tyramine (55 microg/kg) was measured by coronary sinus and aortic catheterization within 1 wk of the PET study. NE levels between coronary sinus and aortic root, DeltaNE(CS-AO), were calculated at baseline and after tyramine administration. Differences of more than 3 SD of baseline (>163 pg/mL) were interpreted as reinnervation. RESULTS: HED retention indicated reinnervation in 10 patients. Maximal HED retention ranged from 4.3%/min to 16.4%/min. DeltaNE(CS AO) 1 min after tyramine administration ranged between -10 pg/mL and 1157 pg/mL, and 8 patients were above the reinnervation threshold. Fisher's exact test demonstrated good agreement between results of PET and DeltaNE(CS-AO) measurements (P = 0.002). Maximal HED retention was also significantly correlated with NE release (r = 0.69; P = 0.001). CONCLUSION: Results of invasively measured NE release and noninvasive (11)C-HED PET are well correlated. This study further supports the usefulness of PET as a noninvasive approach for detection of reappearance of catecholamine uptake sites after heart transplantation. PMID- 11438621 TI - Comparison of cardiac sympathetic nervous function with left ventricular function and perfusion in cardiomyopathies by (123)I-MIBG SPECT and (99m)Tc-tetrofosmin electrocardiographically gated SPECT. AB - The objective of this study was to clarify the relationship between cardiac sympathetic nervous function (CSNF) and left ventricular (LV) function and perfusion in hypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM). METHODS: Thirty-eight cases (32 males, 6 females; mean age, 56 +/- 15 y), consisting of 5 healthy control subjects, 15 patients with DCM, and 18 patients with HCM, were studied with (123)I-metaiodobenzylguanidine (MIBG) and (99m)Tc tetrofosmin SPECT. CSNF was evaluated from cardiac uptake and washout of MIBG, whereas LV perfusion and function were evaluated from tetrofosmin uptake and wall thickening on electrocardiographically gated SPECT. As quantitative parameters of global cardiac MIBG uptake and washout, the heart-to-mediastinum ratio (H/M) and percentage washout were calculated from early and delayed planar images. As quantitative regional parameters, the regional uptake and percentage washout of MIBG were calculated from SPECT images dividing the left ventricle into 12 segments. In the tetrofosmin study, the H/M and LV ejection fraction were calculated as the parameters of global LV perfusion and function. As quantitative regional parameters, the regional uptake and wall thickening were also calculated for the 12 myocardial segments using the quantitative gated SPECT software. Multiple linear regression analysis was performed to investigate the correlations between the parameters from the 2 studies. RESULTS: In DCM and HCM, multiple linear regression analysis of the regional parameters showed significant correlations between LV function and CSNF (P < 0.0001) and between LV perfusion and CSNF (P < 0.0001). According to the partial correlation coefficients, washout and early uptake of MIBG were the most significant factors for predicting LV function and LV perfusion, respectively. CONCLUSION: In cardiomyopathies, CSNF was closely related to LV function. The quantitative parameters of MIBG washout could reflect cardiac functional impairment. Early MIBG uptake might be determined by myocardial perfusion in cardiomyopathies. PMID- 11438622 TI - On the quantification of [18F]MPPF binding to 5-HT1A receptors in the human brain. AB - Previous studies have shown that 4-(2'-methoxyphenyl)-1-[2'-(N-2"-pyridinyl)-p [(18)F]fluorobenzamido]ethylpiperazine ([(18)F]MPPF) binds with high selectivity to serotonin (5-HT(1A)) receptors in man. However, in these studies, the calculation of the binding potential (BP, which equals receptor density divided by equilibrium dissociation constant) used a metabolite-corrected arterial input. The aim of this study was to determine whether metabolite correction and arterial sampling are essential for the assessment of BP. METHODS: Five analytic methods using full datasets obtained from 6 healthy volunteers were compared. In addition, the clinical applicability of these methods was appraised. Three methods were based on Logan analysis of the dynamic PET data using metabolite corrected and uncorrected arterial plasma input and cerebellar input. The other 2 methods consisted of a simplified reference tissue model and standard compartmental modeling. RESULTS: A high correlation was found between BP calculated with Logan analysis using the metabolite-corrected plasma input (used as the reference method for this study) and Logan analysis using either the uncorrected arterial plasma input (r(2) = 0.95, slope = 0.85) or cerebellar input (r(2) = 0.98, slope = 0.91). A high correlation was also found between our reference method and the simplified reference tissue model (r(2) = 0.94, slope = 0.92). In contrast, a poor correlation was observed between our reference method and the standard compartmental model (r(2) = 0.45, slope = 1.59). CONCLUSION: These results indicate that neither metabolite analysis nor arterial sampling is necessary for clinical evaluation of BP in the human brain with [(18)F]MPPF. Both the Logan analysis method with cerebellar input and the simplified reference tissue method can be applied clinically. PMID- 11438623 TI - Effect of vascular radioactivity on regional values of cerebral blood flow: evaluation of methods for H(2)(15)O PET to distinguish cerebral perfusion from blood volume. AB - To evaluate the appropriate model for calculating regional cerebral blood flow (rCBF) with PET and H(2)(15)O, the values obtained from 1- and 2-compartment analyses were compared. METHODS: Dynamic PET scans were performed on 12 healthy volunteers after injection of H(2)(15)O in 2 conditions of baseline and visual stimulation. Calculation of rCBF was performed using the 2-weighted integral (WI) and autoradiographic methods for the 1-compartment analysis, and the 3-WI method was followed for the 2-compartment analysis. Arterial blood radioactivity was counted continuously and corrected for delay and dispersion. The rCBF images were transformed into the Talairach space and analyzed by statistical parametric mapping to identify regional differences in the 2 methods. The values obtained from regions of interest also were compared. RESULTS: Although the difference in global CBF between the 2 models was not significant, rCBF values in the large arteries and neighbor areas were significantly greater in the 2-WI method than in the 3-WI method. However, regional differences in the activation studies were not affected when the 2 methods were compared. The images of cerebral arterial blood volume (V(0)) obtained by the 3-WI method showed a significant increase in V(0) in the visual cortex during visual stimulation. CONCLUSION: These results suggest that the rCBF values in the 1-compartment analysis were affected by radioactivity in the vessels. The 3-WI method could provide rCBF values that are less influenced by vascular radioactivity and also show differences in V(0) in PET activation studies. PMID- 11438624 TI - Distinction between cerebral blood flow and vascular activity by PET tracer kinetic analysis: practical and physiologic considerations. PMID- 11438625 TI - Electrocardiographically gated blood-pool SPECT and left ventricular function: comparative value of 3 methods for ejection fraction and volume estimation. AB - The current major limitation to development of electrocardiographically (ECG) gated blood-pool SPECT (GBPS) for measurement of the left ventricular (LV) ejection fraction (LVEF) and volumes is the lack of availability of clinically validated automatic processing software. Recently, 2 processing software methods for quantification of the LV function have been described. Their LVEFs have been validated separately, but no validation of the LV volume measurement has been reported. METHODS: We compared 3 processing methods for evaluation of the LVEF (n = 29) and volumes (n = 58) in 29 patients: automatic geometric method (GBPS(G)), semiautomatic activity method (GBPS(M)), and 35% maximal activity manual method (GBPS(35%)). The LVEF provided by the ECG gated equilibrium planar left anterior oblique view (planar(LAO)) and the LV volumes provided by LV digital angiography (Rx) were used as gold standards. RESULTS: Whereas the GBPS(G) and GBPS(M) methods present similar low percentage variabilities, the GBPS(35%) method provided the lowest percentage variabilities for the LVEF and volume measurements (P < 0.04 and P < 0.02, respectively). The LVEF and volume provided by the 3 methods were highly correlated with the gold standard methods (r > 0.98 and r > 0.83, respectively). The LVEFs provided by the GBPS(35%) and GBPS(M) methods are similar and higher than those of the GBPS(G) method and planar(LAO) method, respectively (P < 0.0001). For the LVEF, there is no correlation between the average and paired absolute difference for the 3 GBPS methods against the planar(LAO) method, and the limits of agreement are relatively large. LV volumes are lower when calculated with the GBPS(M), GBPS(G), and Rx methods (P < 0.0001). However, the GBPS(35%) and Rx methods provide LV volumes that are similar. There is no linear correlation between the average and the paired absolute difference of volumes calculated with the GBPS(G) and GBPS(35%) methods against Rx LV volumes. However, a moderate linear correlation was found with the GBPS(M) method (r = 0.6; P = 0.0001). The 95% limits of agreement between the Rx LV volumes and the 3 GBPS methods are relatively large. CONCLUSION: GBPS is a simple, highly reproducible, and accurate technique for the LVEF and volume measurement. The reported findings should be considered when comparing results of different methods (GBPS vs. planar(LAO) LVEF; GBPS vs. Rx volume) and results of different GBPS processing methods. PMID- 11438626 TI - Gated cardiac SPECT: has the addition of function to perfusion strengthened the value of myocardial perfusion imaging? PMID- 11438627 TI - PET imaging of somatostatin receptors using [68GA]DOTA-D-Phe1-Tyr3-octreotide: first results in patients with meningiomas. AB - Imaging of somatostatin receptors (SSTRs) using [111In]diethylenetriaminepentaacetic-acid-octreotide (DTPAOC) has proven to be helpful in the differentiation of meningiomas, neurinomas or neurofibromas, and metastases as well as in the follow-up of meningiomas. A drawback of the SPECT method is its limited sensitivity in detecting small meningiomas. Because of PET's increased spatial resolution and its ability to absolutely quantify biodistribution, a PET tracer for SSTR imaging would be desirable. METHODS: 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic-acid-D-Phe1-Tyr3 octreotide (DOTATOC) was labeled using the positron-emitting generator nuclide 68Ga. We acquired dynamic PET images over 120 min after intravenous injection of 175 MBq [68Ga]DOTATOC in 3 patients suffering from 8 meningiomas (WHO I degrees; 7- to 25-mm diameter). Patients' heads had been fixed using individually shaped fiber masks equipped with an external stereotactic localizer system to match PET, CT, and MRI datasets. RESULTS: [68Ga]DOTATOC was rapidly cleared from the blood (half-life alpha, 3.5 min; half-life beta, 63 min). Standardized uptake values (SUVs) of meningiomas increased immediately after injection and reached a plateau 60-120 min after injection (mean SUV, 10.6). No tracer could be found in the surrounding healthy brain tissue. All meningiomas (even the 3 smallest [7- to 8 mm diameter]) showed high tracer uptake and could be visualized clearly. Tracer boundaries showed a good correspondence with the matched CT and MRI images. PET provided valuable additional information regarding the extent of meningiomas located beneath osseous structures, especially at the base of the skull. CONCLUSION: According to our initial experiences, [68Ga]DOTATOC seems to be a very promising new PET tracer for imaging SSTRs even in small meningiomas, offering excellent imaging properties and a very high tumor-to-background ratio. PMID- 11438628 TI - Somatostatin receptor imaging: current status and future perspectives. PMID- 11438629 TI - Occipital brain perfusion deficits in children with major depressive disorder. AB - Occipital lobe perfusion defects have been identified on regional cerebral blood flow (rCBF) SPECT scans of adolescent children and young adults with major depressive disorder (MDD). We reinvestigated a series of rCBF SPECT scans obtained several years ago on drug-naive children with a clinical diagnosis of MDD and on healthy children. METHODS: To test whether visually apparent abnormalities in rCBF constitute statistically significant differences between patients, given the relatively small sample sizes, we applied the technique of statistical parametric mapping (SPM). RESULTS: Two groups of patients were identified: 8 with significant posterior flow deficits in the occipital cortex (Brodmann's areas 18 and 19), usually symmetric, and best visualized on paramedian sagittal sections, and 13 without obvious occipital perfusion deficits but with anterior rCBF deficits in a pattern often described in the literature, attaining statistical significance in the right frontal region. Other localizations in the left frontal and bilateral prefrontal regions did not attain significance, but each localization contained statistically significant maxima (z scores). The scan findings of all 18 healthy children were normal. CONCLUSION: With the aid of SPM, 2 groups of children with significantly different rCBF behavior were identified. The reason for this difference is not known but should be investigated to determine its possible significance to patients with MDD. PMID- 11438630 TI - Cardiac neurotransmission imaging. AB - Cardiac neurotransmission imaging with SPECT and PET allows in vivo assessment of presynaptic reuptake and neurotransmitter storage as well as of regional distribution and activity of postsynaptic receptors. In this way, the biochemical processes that occur during neurotransmission can be investigated in vivo at a micromolar level using radiolabeled neurotransmitters and receptor ligands. SPECT and PET of cardiac neurotransmission characterize myocardial neuronal function in primary cardioneuropathies, in which the heart has no significant structural abnormality, and in secondary cardioneuropathies caused by the metabolic and functional changes that take place in different diseases of the heart. In patients with heart failure, the assessment of sympathetic activity has important prognostic implications and will result in better therapy and outcome. In diabetic patients, scintigraphic techniques allow the detection of autonomic neuropathy in early stages of the disease. In conditions with a risk of sudden death, such as idiopathic ventricular tachycardia and arrhythmogenic right ventricular cardiomyopathy, PET and SPECT reveal altered neuronal function when no other structural abnormality is seen. In patients with ischemic heart disease, heart transplantation, drug-induced cardiotoxicity, and dysautonomias, assessment of neuronal function can help characterize the disease and improve prognostic stratification. Future directions include the development of tracers for new types of receptors, the targeting of second messenger molecules, and the early assessment of cardiac neurotransmission in genetically predisposed subjects for prevention and early treatment of heart failure. PMID- 11438631 TI - Usefulness and pitfalls of planar gamma-scintigraphy for measuring aerosol deposition in the lungs: a Monte Carlo investigation. AB - Planar gamma-scintigraphy is often used to quantify pulmonary deposition patterns from aerosol inhalers. The results are quite different from those obtained using 3-dimensional PET and SPECT. The purpose of this study was to characterize the effects of scatter and tissue attenuation on the distribution of radiolabeled aerosol as measured by planar scintigraphy using Monte Carlo simulations. This study also investigated the applicability of a few correction methods used in inhalation studies. METHODS: Body density maps were derived from CT scans. Regions of interest-lungs, major airways, and esophagus-were defined from the same CT volume. Two radioactivity source distribution patterns in the lung, uniform and nonuniform, were used. A Monte Carlo program, SIMIND, was used to generate anterior and posterior gamma-images of the composed inhalation distributions for 2 energy windows, photopeak (127-153 keV) and scatter (92-125 keV). The effects of scatter and attenuation were estimated on the basis of the imaging components separated from the simulation. A scatter correction method and 2 attenuation correction methods, all applied to inhalation scintigraphy, were evaluated using the simulated images. RESULTS: The amount of scatter ranges from 24% to approximately 29% in the lungs and from 29% to approximately 35% in the central (airway or esophagus) region on the planar images. Significant differences were found among regions and between source distributions (P < 0.05). The fraction k used for dual-energy-based scatter correction also varied and was found to be less than the commonly used k = 0.5. The simplified narrow-beam attenuation correction and the effective (broad-beam) correction methods were found to either under- or overcorrect the regional activities. CONCLUSION: The amount of scatter and tissue attenuation in the thorax region depends on source distribution and body attenuation. In applying planar scintigraphy for aerosol inhalation studies, it is difficult to obtain precise quantitative measurements because of the uncertainties associated with scatter and attenuation corrections. Accurate corrections require knowledge of both source and density distributions. PMID- 11438632 TI - Improved prediction of therapeutic absorbed doses of radioiodine in the treatment of thyroid carcinoma. AB - We proposed an alternative to a monoexponential model of radioiodine kinetics to obtain a more accurate estimate of absorbed doses to postsurgical thyroid remnants. We suggested that part of the difference between the predicted and the actually absorbed therapeutic doses of (131)I, usually explained by radiation damage of thyroid cells, can be attributed to errors resulting from inadequate sampling of data and oversimplified modeling. METHODS: A standard monoexponential model and alternative biphasic model (incorporating both radioiodine uptake and clearance) were used on 2 sets of patient data to fit time-activity measurements after administration of diagnostic and therapeutic activities of radioiodine. One set of data consisted of 633 records of routine measurements, and the second set consisted of 71 prospectively collected records with measurements performed more frequently and for a longer time. The time-activity curves derived from the 2 models were used to calculate residence times for diagnostic and therapeutic activities of (131)I, and the respective residence times were compared using the paired t test. Errors of fitting and prediction of therapeutic time-activity data were also calculated. RESULTS: With both models, a statistically significant difference (P < 0.01) was found between residence times after diagnostic administration of (131)I and residence times after therapeutic administration of (131)I. However, the effects of biphasic modeling and of improved sampling substantially reduced the difference (P < 0.01). Errors of fitting and prediction were smaller with the biphasic model than with the monoexponential model (P < 0.01). CONCLUSION: The biphasic model more accurately predicts (131)I kinetics when applied to measurements in the short interval after diagnostic administration of radioiodine. The minimum requirement for the biphasic model is measurement twice a day at intervals > 6 h for at least 3 d after administration. PMID- 11438633 TI - Assessment of porcine bone metabolism by dynamic. AB - The aim of this study was to quantify regional bone blood flow and [(18)F]fluoride ion influx with [(18)F]fluoride ion PET and correlate the results with specific static and dynamic indices of bone metabolism in healthy pigs. METHODS: During continuous ventilation (fractional concentration of oxygen in inspired gas = 0.3), dynamic PET scans 120 min in duration were obtained for 9 mini pigs after intravenous injection of 10.0 +/- 1.2 MBq (mean +/- SD) of [(18)F]fluoride ion per kilogram of body weight. Iliac crest bone biopsies were performed immediately before the PET scan to determine static and dynamic indices of bone metabolism (i.e., the mineral apposition rate) by bone histomorphometry. Kinetic rate constants describing influx (K(1)) and efflux (k(2)) of [(18)F]fluoride as well as chemisorption and incorporation of [(18)F]fluoride (k(3)) and reverse transport (k(4)) were determined for 6 vertebral bodies in each animal. Blood flow estimates (f) were derived from K(1) values corrected for the permeability-surface area product using a previously derived correction algorithm. A rate constant describing the net forward transport rate of fluoride (K(i)) and the fluoride volume flux (K(flux)) derived from a 2-tissue-compartment model was calculated and compared with the results of Patlak graphic analysis (K(pat)). RESULTS: A significant correlation was found between mineral apposition rate and K(i) (P < 0.005), K(flux) (P < 0.01), K(pat), K(1), and f (P < 0.05). The values of f, K(i), K(flux), and K(pat) did not correlate significantly with other static or dynamic histomorphometric indices or with age, serum alkaline phosphatase, or parathyroid hormone levels. The values of f and K(i) correlated linearly (y = 0.023 + 0.32x; r(2) = 0.74; P < 0.001). CONCLUSION: PET bone studies using [(18)F]fluoride ion provide quantitative estimates of bone blood flow and metabolic activity that correlate with histomorphometric indices of bone formation in the normal bone tissue of the mini pig. Therefore, it seem reasonable to assume that [(18)F]fluoride ion PET can reduce the number of invasive bone biopsies, thus facilitating follow-up of patients with metabolic bone diseases. PMID- 11438634 TI - Extraction of long-chain fatty acids in isolated rat heart during acute low-flow ischemia. AB - Although beta-oxidation of fatty acids is suppressed rapidly during ischemia, the behavior of fatty acid extraction at different flow rates is incompletely understood. This study assessed the relationship between flow and extraction of (123)I-iodophenylpentadecanoic acid (IPPA) in the isolated heart model, especially at low flow. METHODS: Isolated hearts from male Wistar rats (n = 15) were subjected to retrograde perfusion with constant flow (Krebs Henseleit solution containing 10 mmol/L glucose). A latex balloon in the left ventricle allowed isovolumetric contractions and ventricular pressure measurements. The extraction of (123)I-IPPA was assessed with the indicator dilution technique and (99m)Tc-albumin as the intravascular reference. The flow was either increased from the control flow (8 mL/min) until 300% or reduced until 10%. (123)I-IPPA extraction was measured three times before and 10 min after flow alteration. The tracer uptake was estimated from the product of net extraction and flow. RESULTS: The mean (123)I-IPPA extraction at the control flow (third measurement) was 51.6% +/- 2.8%. Between flow rates of approximately 25% and 300%, (123)I-IPPA extraction increased exponentially at decreasing flow rates. At flow rates < or =25% of the control flow, (123)I-IPPA extraction was exponentially higher than predicted. (123)I-IPPA uptake and flow changed largely in parallel. During low flow, the rate-pressure product showed the expected decline (perfusion contraction matching). CONCLUSION: The extraction of (123)I-IPPA is preserved and slightly increased (relative to flow) during acute low-flow ischemia. PMID- 11438635 TI - Early detection of oleic acid-induced lung injury in rats using (111)In-labeled anti-rat intercellular adhesion molecule-1. AB - Previous study of the bleomycin-induced lung injury model suggested that (111)In labeled antirat intercellular adhesion molecule-1 (aICAM-1) might be a useful acute respiratory distress syndrome (ARDS) diagnostic agent. We further investigated the ability of (111)In-aICAM-1 to detect inflammation in another ARDS lung injury model. METHODS: (111)In-labeled rat polymorphonuclear leukocytes (PMNs), (111)In-aICAM-1, (111)In-labeled normal mouse IgG (nmIgG), and (111)In labeled rat serum albumin (RSA) were injected into rats 18-24 h before kill. Biodistributions, scintigraphic images, and lung ICAM-1 upregulation were obtained in uninjured rats and in rats after injury with oleic acid. RESULTS: (111)In-RSA and (111)In-nmIgG localized in inflamed lung at 5 min postinjury (PI). (111)In-PMN uptake increased significantly only at 24 h PI. (111)In-aICAM-1 localization increased significantly (30%-60%) at 1 h PI and remained elevated up to 24 h PI. Lung/blood ratios (L/B) at 1 and 4 h PI were very low (<0.6) for (111)In-nmIgG and (111)In-PMN rats; however, for (111)In-aICAM-1 rats, they were >1 and 25%-60% higher than those for the control samples. A low L/B suggests poor inflammation detection on the images. Images and region-of-interest analysis confirmed that only (111)In-aICAM-1 could distinguish inflamed lungs at 4 h PI. ICAM-1 was upregulated at 4 and 24 h PI. CONCLUSION: In this model, (111)In-aICAM 1 detected lung inflammation very early in the course of the disease. These results support the suggestion that (111)In-aICAM-1 could be a very early, highly specific ARDS diagnostic agent and may be useful to detect a wide range of inflammations. PMID- 11438636 TI - Evaluation of geometric sensitivity for hybrid PET. AB - Hybrid PET systems have spatially varying sensitivity profiles. These profiles are dependent on imaging parameters, namely, number of heads, head configuration, spacing between gantry stops, radius of rotation (RoR), and coincident head acceptance angle. METHODS: Sensitivity profiles were calculated across a 500-mm field of view (FoV) for a representative set of existing and theoretic 2-, 3-, and 4-head hybrid PET systems. The head configuration was defined by alpha(n), which describes the angular separation between head 1 and head n. Simulated configurations were 2 head ([alpha(2)] = [180 degrees ]), 3 head ([alpha(2), alpha(3)] = [120 degrees, 240 degrees ] and [90 degrees, 180 degrees ]), and 4 head ([alpha(2), alpha(3), alpha(4)] = [90 degrees, 180 degrees, 270 degrees ]). Four transverse acceptance angles, measured from the perpendicular of the crystal to the surface, were simulated: 90 degrees, 45 degrees, 23 degrees, and 11 degrees. Two RoRs were considered: 250 and 300 mm. Each head was rotated through 360 degrees in 128 steps, and no physical collimation was modeled. RESULTS: For a 250-mm RoR and 90 degrees acceptance angle, the sensitivities relative to [alpha(2)] = [180 degrees ] were [alpha(2), alpha(3)] = [120 degrees, 240 degrees ], 183%; [alpha(2), alpha(3)] = [90 degrees, 180 degrees ], 159%; and [alpha(2), alpha(3), alpha(4)] = [90 degrees, 180 degrees, 270 degrees ], 317%. Increasing RoR to 300 mm decreased [alpha(2)] = [180 degrees ] sensitivity by approximately 12%; all other configurations were decreased by approximately 75% of their 250-mm RoR sensitivities. Decreasing the acceptance angle to 45 degrees decreased sensitivities to [alpha(2), alpha(3)] = [120 degrees, 240 degrees ], 100%; [alpha(2), alpha(3)] = [90 degrees, 180 degrees ], 105%; and [alpha(2), alpha(3), alpha(4)] = [90 degrees, 180 degrees, 270 degrees ], 210%. The 2-head [alpha(2)] = [180 degrees ] system sensitivity was not affected. The configuration was the most important factor affecting the shape of the sensitivity profiles. For a 250 mm RoR and 90 degrees acceptance angle, [alpha(2)] = [180 degrees ] concentrated sensitivity in the FoV center, [alpha(2), alpha(3)] = [120 degrees, 240 degrees ] had a slightly increased peripheral sensitivity, and the profiles for both [alpha(2), alpha(3)] = [90 degrees, 180 degrees ] and [alpha(2), alpha(3), alpha(4)] = [90 degrees, 180 degrees, 270 degrees ] were completely flat. CONCLUSION: Sensitivity profiles are affected strongly by imaging parameters; however, profiles can be shaped to concentrate on an annulus or distribute sensitivity uniformly over the FoV. Also, the 4-head system showed a markedly higher sensitivity than either of the 3-head systems. PMID- 11438637 TI - Emission tuned-aperture computed tomography: a novel approach to scintimammography. AB - Emission tuned-aperture computed tomography (ETACT) is a new approach to acquiring and processing scintimammography data. A gamma camera with a pinhole collimator is used to acquire projections of the radionuclide distribution within the breast. Fiducial markers are used to reconstruct these projections into tomographic slices. Simulation and phantom experiments were performed to evaluate the potential of the ETACT method. METHODS: In the simulation study, a hemispheric object of 15 cm in diameter was constructed to model a breast. A ray tracing technique was used to generate ideal projections. These were blurred and noise was added to create images that resemble scintigraphic images. Tumor size, pinhole size, and target-to-nontarget radioactivity ratios (TNTs) were varied. The simulated projections were reconstructed into slices, and contrast and contrast-to-noise ratios were calculated to evaluate the effect of pinhole size. These results were compared with a simulated planar acquisition of the same object. A preliminary phantom evaluation was performed using an 8-mm "tumor" with a 10:1 TNT to validate the simulation results. RESULTS: A 3-mm pinhole was shown by the simulation study to be the optimal size. The ETACT images consistently yielded higher contrast than simulated planar images. The phantom study validated the simulation results and showed the feasibility of ETACT in a simulated clinical environment. CONCLUSION: ETACT is shown to be useful for imaging tumors <1 cm in diameter. Because ETACT requires only a gamma camera with a pinhole collimator, it has the potential to be applied in any hospital in a simple, flexible, and practical manner. PMID- 11438638 TI - Thyroid stunning. PMID- 11438639 TI - Stunning after tracer dosimetry. PMID- 11438640 TI - SPECT imaging in dementias. PMID- 11438641 TI - Procedure guideline for somatostatin receptor scintigraphy with (111)In pentetreotide. PMID- 11438642 TI - New model for the yeast RNA polymerase I transcription cycle. AB - Using an immobilized template assay, we observed two steps in assembly of the yeast RNA polymerase I (Pol I) preinitiation complex: stable binding of upstream activating factor (UAF) followed by recruitment of Pol I-Rrn3p and core factor (CF). Pol I is required for stable association of CF with the promoter and can be recruited in the absence of Rrn3p. Upon transcription initiation, Pol I-Rrn3p and CF dissociate from the promoter while UAF remains behind. These findings support a novel model in which the Pol I basal machinery cycles on and off the promoter with each round of transcription. This model accounts for previous observations that rRNA synthesis may be controlled by regulating both promoter accessibility and polymerase activity. PMID- 11438643 TI - Transcription factor AP-2alpha is preferentially cleaved by caspase 6 and degraded by proteasome during tumor necrosis factor alpha-induced apoptosis in breast cancer cells. AB - Several reports have linked activating protein 2alpha (AP-2alpha) to apoptosis, leading us to hypothesize that AP-2alpha is a substrate for caspases. We tested this hypothesis by examining the effects of tumor necrosis factor alpha (TNF alpha) on the expression of AP-2 in breast cancer cells. Here, we provide evidence that TNF-alpha downregulates AP-2alpha and AP-2gamma expression posttranscriptionally during TNF-alpha-induced apoptosis. Both a general caspase antagonist (zVADfmk) and a caspase 6-preferred antagonist (zVEIDfmk) inhibited TNF-alpha-induced apoptosis and AP-2alpha downregulation. In vivo tests showed that AP-2alpha was cleaved by caspases ahead of the DNA fragmentation phase of apoptosis. Recombinant caspase 6 cleaved AP-2alpha preferentially, although caspases 1 and 3 also cleaved it, albeit at 50-fold or higher concentrations. Activated caspase 6 was detected in TNF-alpha-treated cells, thus confirming its involvement in AP-2alpha cleavage. All three caspases cleaved AP-2alpha at asp(19) of the sequence asp-arg-his-asp (DRHD(19)). Mutating D(19) to A(19) abrogated AP-2alpha cleavage by all three caspases. TNF-alpha-induced cleavage of AP-2alpha in vivo led to AP-2alpha degradation and loss of DNA-binding activity, both of which were prevented by pretreatment with zVEIDfmk. AP-2alpha degradation but not cleavage was inhibited in vivo by PS-431 (a proteasome antagonist), suggesting that AP-2alpha is degraded subsequent to cleavage by caspase 6 or caspase 6-like enzymes. Cells transfected with green fluorescent protein-tagged mutant AP-2alpha are resistant to TNF-alpha-induced apoptosis, further demonstrating the link between caspase-mediated cleavage of AP-2alpha and apoptosis. This is the first report to demonstrate that degradation of AP-2alpha is a critical event in TNF-alpha-induced apoptosis. Since the DRHD sequence in vertebrate AP-2 is widely conserved, its cleavage by caspases may represent an important mechanism for regulating cell survival, proliferation, differentiation, and apoptosis. PMID- 11438644 TI - Mutational analysis of the Cy motif from p21 reveals sequence degeneracy and specificity for different cyclin-dependent kinases. AB - Inhibitors, activators, and substrates of cyclin-dependent kinases (cdks) utilize a cyclin-binding sequence, known as a Cy or RXL motif, to bind directly to the cyclin subunit. Alanine scanning mutagenesis of the Cy motif of the cdk inhibitor p21 revealed that the conserved arginine or leucine (constituting the conserved RXL sequence) was important for p21's ability to inhibit cyclin E-cdk2 activity. Further analysis of mutant Cy motifs showed, however, that RXL was neither necessary nor sufficient for a functional cyclin-binding motif. Replacement of either of these two residues with small hydrophobic residues such as valine preserved p21's inhibitory activity on cyclin E-cdk2, while mutations in either polar or charged residues dramatically impaired p21's inhibitory activity. Expressing p21N with non-RXL Cy sequences inhibited growth of mammalian cells, providing in vivo confirmation that RXL was not necessary for a functional Cy motif. We also show that the variant Cy motifs identified in this study can effectively target substrates to cyclin-cdk complexes for phosphorylation, providing additional evidence that these non-RXL motifs are functional. Finally, binding studies using p21 Cy mutants demonstrated that the Cy motif was essential for the association of p21 with cyclin E-cdk2 but not with cyclin A-cdk2. Taking advantage of this differential specificity toward cyclin E versus cyclin A, we demonstrate that cell growth inhibition was absolutely dependent on the ability of a p21 derivative to inhibit cyclin E-cdk2. PMID- 11438645 TI - Multifunctional centromere binding factor 1 is essential for chromosome segregation in the human pathogenic yeast Candida glabrata. AB - The CBF1 (centromere binding factor 1) gene of Candida glabrata was cloned by functional complementation of the methionine biosynthesis defect of a Saccharomyces cerevisiae cbf1 deletion mutant. The C. glabrata-coded protein, CgCbf1, contains a basic-helix-loop-helix leucine zipper domain and has features similar to those of other budding yeast Cbf1 proteins. CgCbf1p binds in vitro to the centromere DNA element I (CDEI) sequence GTCACATG with high affinity (0.9 x 10(9) M(-1)). Bandshift experiments revealed a pattern of protein-DNA complexes on CgCEN DNA different from that known for S. cerevisiae. We examined the effect of altering the CDEI binding site on CEN plasmid segregation, using a newly developed colony-sectoring assay. Internal deletion of the CDEI binding site led only to a fivefold increase in rates of plasmid loss, indicating that direct binding of Cbf1p to the centromere DNA is not required for full function. Additional deletion of sequences to the left of CDEI, however, led to a 70-fold increase in plasmid loss rates. Deletion of the CBF1 gene proved to be lethal in C. glabrata. C. glabrata cells containing the CBF1 gene under the influence of a shutdown promoter (tetO-ScHOP) arrested their growth after 5 h of cultivation in the presence of the reactive drug doxycycline. DAPI (4',6'-diamidino-2 phenylindole) staining of the arrested cells revealed a significant increase in the number of large-budded cells with single nuclei, 2C DNA content, and short spindles, indicating a defect in the G(2)/M transition of the cell cycle. Thus, we conclude that Cbf1p is required for chromosome segregation in C. glabrata. PMID- 11438646 TI - Identification of rad27 mutations that confer differential defects in mutation avoidance, repeat tract instability, and flap cleavage. AB - In eukaryotes, the nuclease activity of Rad27p (Fen1p) is thought to play a critical role in lagging-strand DNA replication by removing ribonucleotides present at the 5' ends of Okazaki fragments. Genetic analysis of Saccharomyces cerevisiae also has identified a role for Rad27p in mutation avoidance. rad27Delta mutants display both a repeat tract instability phenotype and a high rate of forward mutations to canavanine resistance that result primarily from duplications of DNA sequences that are flanked by direct repeats. These observations suggested that Rad27p activities in DNA replication and repair could be altered by mutagenesis and specifically assayed. To test this idea, we analyzed two rad27 alleles, rad27-G67S and rad27-G240D, that were identified in a screen for mutants that displayed repeat tract instability and mutator phenotypes. In chromosome stability assays, rad27-G67S strains displayed a higher frequency of repeat tract instabilities relative to CAN1 duplication events; in contrast, the rad27-G240D strains displayed the opposite phenotype. In biochemical assays, rad27-G67Sp displayed a weak exonuclease activity but significant single- and double-flap endonuclease activities. In contrast, rad27 G240Dp displayed a significant double-flap endonuclease activity but was devoid of exonuclease activity and showed only a weak single-flap endonuclease activity. Based on these observations, we hypothesize that the rad27-G67S mutant phenotypes resulted largely from specific defects in nuclease function that are important for degrading bubble intermediates, which can lead to DNA slippage events. The rad27-G240D mutant phenotypes were more difficult to reconcile to a specific biochemical defect, suggesting a structural role for Rad27p in DNA replication and repair. Since the mutants provide the means to relate nuclease functions in vitro to genetic characteristics in vivo, they are valuable tools for further analyses of the diverse biological roles of Rad27p. PMID- 11438647 TI - Linking the 3' poly(A) tail to the subunit joining step of translation initiation: relations of Pab1p, eukaryotic translation initiation factor 5b (Fun12p), and Ski2p-Slh1p. AB - The 3' poly(A) structure improves translation of a eukaryotic mRNA by 50-fold in vivo. This enhancement has been suggested to be due to an interaction of the poly(A) binding protein, Pab1p, with eukaryotic translation initiation factor 4G (eIF4G). However, we find that mutation of eIF4G eliminating its interaction with Pab1p does not diminish the preference for poly(A)(+) mRNA in vivo, indicating another role for poly(A). We show that either the absence of Fun12p (eIF5B), or a defect in eIF5, proteins involved in 60S ribosomal subunit joining, specifically reduces the translation of poly(A)(+) mRNA, suggesting that poly(A) may have a role in promoting the joining step. Deletion of two nonessential putative RNA helicases (genes SKI2 and SLH1) makes poly(A) dispensable for translation. However, in the absence of Fun12p, eliminating Ski2p and Slh1p shows little enhancement of expression of non-poly(A) mRNA. This suggests that Ski2p and Slh1p block translation of non-poly(A) mRNA by an effect on Fun12p, possibly by affecting 60S subunit joining. PMID- 11438648 TI - Ligand-dependent degradation of retinoid X receptors does not require transcriptional activity or coactivator interactions. AB - Cells utilize ubiquitin-mediated proteolysis to regulate the activity of numerous proteins involved in signal transduction, cell cycle control, and transcriptional regulation. For a number of transcription factors, there appears to be a direct correlation between transcriptional activity and protein instability, suggesting that cells use targeted destruction as one method to down-regulate or attenuate gene expression. In this report we demonstrate that retinoid X receptors (RXRs) which function as versatile mediators of nuclear hormone-dependent gene expression are marked for destruction upon binding agonist ligands. Interestingly, when RXR serves as a heterodimeric partner for retinoic acid (RAR) or thyroid hormone (TR) receptors, binding of agonists by RAR or TR leads to degradation of both the transcriptionally active RAR or TR subunits as well as the transcriptionally inactive RXR subunit. Furthermore, using a series of mutants in the ligand-dependent activation domain (activation function 2), we demonstrate that agonist-stimulated degradation of RXR does not require corepressor release, coactivator binding, or transcriptional activity. Taken together, the data suggest a model for targeted destruction of transcription factors based on structural or conformational signals as opposed to functional coupling with gene transcription. PMID- 11438649 TI - Self-association of CIITA and its transactivation potential. AB - The major histocompatibility complex (MHC) class II transactivator (CIITA) regulates the expression of genes involved in the immune response, including MHC class II genes and the interleukin-4 gene. Interactions between CIITA and sequence-specific, DNA-binding proteins are required for CIITA to function as an activator of MHC class II genes. CIITA also interacts with the coactivators CBP (also called p300), and this interaction leads to synergistic activation of MHC class II promoters. Here, we report that CIITA forms complexes with itself and that a central region, including the GTP-binding domain is sufficient for self association. Additionally, this central region interacts with the C-terminal leucine-rich repeat as well as the N-terminal acidic domain. LXXLL motifs residing in the GTP-binding domain are essential for self-association. Finally, distinct differences exist among various CIITA mutant proteins with regard to activation function, subcellular localization, and association with wild-type protein and dominant-negative potential. PMID- 11438650 TI - c-Myc is necessary for DNA damage-induced apoptosis in the G(2) phase of the cell cycle. AB - The c-myc proto-oncogene encodes a transcription factor that participates in the regulation of cellular proliferation, differentiation, and apoptosis. Ectopic overexpression of c-Myc has been shown to sensitize cells to apoptosis. We report here that cells lacking c-Myc activity due to disruption of the c-myc gene by targeted homologous recombination are defective in DNA damage-initiated apoptosis in the G(2) phase of the cell cycle. The downstream effector of c-Myc is cyclin A, whose ectopic expression in c-myc(-/-) cells rescues the apoptosis defect. The kinetics of the G(2) response indicate that the induction of cyclin A and the concomitant activation of Cdk2 represent an early step during commitment to apoptosis. In contrast, expression of cyclins E and D1 does not rescue the apoptosis defect, and apoptotic processes in G(1) phase are not affected in c myc(-/-) cells. These observations link DNA damage-induced apoptosis with cell cycle progression and implicate c-Myc in the functioning of a subset of these pathways. PMID- 11438651 TI - DNA replication forks pause at silent origins near the HML locus in budding yeast. AB - Chromosomal replicators in budding yeast contain an autonomously replicating sequence (ARS) that functions in a plasmid, but certain ARSs are silent as replication origins in their natural chromosomal context. In chromosome III, the HML ARS cluster (ARS302-ARS303-ARS320) and ARS301 flank the transcriptionally silent mating-type locus HML, and all of these ARSs are silent as replication origins. ARS301 and ARS302 function in transcriptional silencing mediated by the origin recognition complex (ORC) and a heterochromatin structure, while the functions of ARS303 and ARS320 are not known. In this work, we discovered replication fork pause sites at the HML ARS cluster and ARS301 by analyzing DNA replication intermediates from the chromosome via two-dimensional gel electrophoresis. The replication fork pause at the HML ARS cluster was independent of cis- and trans-acting mutations that abrogate transcriptional silencing at HML. Deletion of the HML ARS cluster led to loss of the pause site. Insertion of a single, heterologous ARS (ARS305) in place of the HML ARS cluster reconstituted the pause site, as did multiple copies of DNA elements (A and B1) that bind ORC. The orc2-1 mutation, known to alter replication timing at origins, did not detectably affect the pause but activated the silent origin at the HML ARS cluster in a minority of cells. Delaying the time of fork arrival at HML led to the elimination of the pause sites at the HML ARS cluster and at the copy of ARS305 inserted in place of the cluster. Loss of the pause sites was accompanied by activation of the silent origins in the majority of cells. Thus, replication fork movement near HML pauses at a silent origin which is competent for replication initiation but kept silent through Orc2p, a component of the replication initiator. Possible functions for replication fork pause sites in checkpoints, S-phase regulation, mating-type switching, and transcriptionally silent heterochromatin are discussed. PMID- 11438652 TI - Cdc5 interacts with the Wee1 kinase in budding yeast. AB - Development of a multicellular organism requires that mitosis and morphogenesis be coordinated. These processes must also be synchronized during the growth of unicellular organisms. In the yeast Saccharomyces cerevisiae, mitosis is dependent on the prior growth of a daughter cell in the form of a bud. Overexpression of wild-type Polo-like kinase Cdc5 or a catalytically inactive form resulted in the formation of multinucleate cells in budding yeast. Immunofluorescence analysis of these multinulceate cells showed that mitosis and bud formation were no longer linked. Others have shown that Swe1 is required for coupling mitosis to bud formation during a perturbed cell cycle. When the normal pathway of bud formation is perturbed, Swe1 functions to delay mitosis through negative regulation of Clb/Cdk. In cells lacking Swe1, multinucleate cells are formed in response to delays in bud formation. Affinity purification, two-hybrid analysis, and mutant characterization results suggested that Cdc5 and Swe1 interact. From these results, we conclude that multinucleate formation in response to Cdc5 overexpression is linked to titration of Swe1 function. These results also suggest that Cdc5 may be a negative regulator of Swe1. PMID- 11438653 TI - Control of cyclin-dependent kinase inhibitor p27 expression by cap-independent translation. AB - p27 is a key regulator of cell proliferation through inhibition of G(1) cyclin dependent kinase (CDK) activity. Translation of the p27 mRNA is an important control mechanism for determining cellular levels of the inhibitor. Nearly all eukaryotic mRNAs are translated through a mechanism involving recognition of the 5' cap by eukaryotic initiation factor 4E (eIF4E). In quiescent cells eIF4E activity is repressed, leading to a global decline in translation rates. In contrast, p27 translation is highest during quiescence, suggesting that it escapes the general repression of translational initiation. We show that the 5' untranslated region (5'-UTR) of the p27 mRNA mediates cap-independent translation. This activity is unaffected by conditions in which eIF4E is inhibited. In D6P2T cells, elevated cyclic AMP levels cause a rapid withdrawal from the cell cycle that is correlated with a striking increase in p27. Under these same conditions, cap-independent translation from the p27 5'-UTR is enhanced. These results indicate that regulation of internal initiation of translation is an important determinant of p27 protein levels. PMID- 11438654 TI - The multisubstrate adapter Gab1 regulates hepatocyte growth factor (scatter factor)-c-Met signaling for cell survival and DNA repair. AB - Hepatocyte growth factor (scatter factor) (HGF/SF) is a pleiotrophic mediator of epithelial cell motility, morphogenesis, angiogenesis, and tumorigenesis. HGF/SF protects cells against DNA damage by a pathway from its receptor c-Met to phosphatidylinositol 3-kinase (PI3K) to c-Akt, resulting in enhanced DNA repair and decreased apoptosis. We now show that protection against the DNA-damaging agent adriamycin (ADR; topoisomerase IIalpha inhibitor) requires the Grb2-binding site of c-Met, and overexpression of the Grb2-associated binder Gab1 (a multisubstrate adapter required for epithelial morphogenesis) inhibits the ability of HGF/SF to protect MDCK epithelial cells against ADR. In contrast to Gab1 and its homolog Gab2, overexpression of c-Cb1, another multisubstrate adapter that associates with c-Met, did not affect protection. Gab1 blocked the ability of HGF/SF to cause the sustained activation of c-Akt and c-Akt signaling (FKHR phosphorylation). The Gab1 inhibition of sustained c-Akt activation and of cell protection did not require the Gab1 pleckstrin homology or SHP2 phosphatase binding domain but did require the PI3K-binding domain. HGF/SF protection of parental MDCK cells was blocked by wortmannin, expression of PTEN, and dominant negative mutants of p85 (regulatory subunit of PI3K), Akt, and Pak1; the protection of cells overexpressing Gab1 was restored by wild-type or activated mutants of p85, Akt, and Pak1. These findings suggest that the adapter Gab1 may redirect c-Met signaling through PI3K away from a c-Akt/Pak1 cell survival pathway. PMID- 11438655 TI - Analysis of smu-1, a gene that regulates the alternative splicing of unc-52 pre mRNA in Caenorhabditis elegans. AB - Mutations in the smu-1 gene of Caenorhabditis elegans were previously shown to suppress mutations in the genes mec-8 and unc-52. mec-8 encodes a putative RNA binding protein that affects the accumulation of specific alternatively spliced mRNA isoforms produced by unc-52 and other genes. unc-52 encodes a set of basement membrane proteins, homologs of mammalian perlecan, that are important for body wall muscle assembly and attachment to basement membrane, hypodermis, and cuticle. We show that a presumptive null mutation in smu-1 suppresses nonsense mutations in exon 17 but not exon 18 of unc-52 and enhances the phenotype conferred by an unc-52 splice site mutation in intron 16. We have used reverse transcription-PCR and RNase protection to show that loss-of-function smu 1 mutations enhance accumulation in larvae of an alternatively spliced isoform that skips exon 17 but not exon 18 of unc-52. We have identified smu-1 molecularly; it encodes a nuclearly localized protein that contains five WD motifs and is ubiquitously expressed. The SMU-1 amino acid sequence is more than 60% identical to a predicted human protein of unknown function. We propose that smu-1 encodes a trans-acting factor that regulates the alternative splicing of the pre-mRNA of unc-52 and other genes. PMID- 11438656 TI - NoBP, a nuclear fibroblast growth factor 3 binding protein, is cell cycle regulated and promotes cell growth. AB - Secreted and nuclear forms of fibroblast growth factor 3 (FGF3) have opposing effects on cells. The secreted form stimulates cell growth and transformation, while the nuclear form inhibits DNA synthesis and cell proliferation. By using the yeast two-hybrid system we have identified a nucleolar FGF3 binding protein (NoBP) which coimmunoprecipitated and colocalized with FGF3 in transfected COS-1 cells. Characterization of the NoBP binding domain of FGF3 exactly matched the sequence requirements of FGF3 for its translocation into the nucleoli, suggesting that NoBP might be the nucleolar binding partner of FGF3 essential for its nucleolus localization. Carboxyl-terminal domains of NoBP contain linear nuclear and nucleolar targeting motifs which are capable of directing a heterologous protein beta-galactosidase to the nucleus and the nucleoli. While NoBP expression was detected in all analyzed proliferating established cell lines, NoBP transcription was rapidly downregulated in the promyelocytic leukemia cell line HL60 when induced to differentiate. Analysis on the expression pattern of NoBP mRNA throughout the cell cycle in HeLa cells synchronized by lovastatin demonstrated a substantial upregulation during the late G(1)/early S phase. NoBP overexpression conferred a proliferating effect onto NIH 3T3 cells and can counteract the inhibitory effect of nuclear FGF3, suggesting a role of NoBP in controlling proliferation in cells. We propose that NoBP is the functional target of nuclear FGF3 action. PMID- 11438657 TI - Investigation of elements sufficient to imprint the mouse Air promoter. AB - Imprinted maternal-allele-specific expression of the mouse insulin-like growth factor type 2 receptor (Igf2r) gene depends on a 3.7-kb element named region 2, located in the second intron of the gene. Region 2 carries a maternal-allele specific methylation imprint and contains an imprinted CpG island promoter (Air) that expresses a noncoding antisense RNA from the paternal inherited allele only. Here, we use transgenes to test the minimal requirements for imprinting of Air and to test if the action of region 2 is restricted to Igf2r. Transgenes up to 9 kb with Air as a single promoter are expressed but not imprinted. When coupled to the Igf2r CpG island promoter on a 44-kb transgene, Air was imprinted in one of three lines. However, Air on a 4.6-kb fragment is also imprinted in 2 of 14 lines when inserted in an intron of an adenine phosphoribosyltransferase (Aprt) transgene, and in one line, the imprinted methylation and expression of Air have been transferred onto the Aprt CpG island promoter. These data suggest that a dual CpG island promoter setting may facilitate Air imprinting as a short transgene and also show that Air can transfer imprinting onto other genes. However, for reliable Air imprinting, elements are necessary that are located outside a 44-kb region spanning the Air-Igf2r promoters. PMID- 11438658 TI - Tight binding of the phosphorylated alpha subunit of initiation factor 2 (eIF2alpha) to the regulatory subunits of guanine nucleotide exchange factor eIF2B is required for inhibition of translation initiation. AB - Translation initiation factor 2 (eIF2) is a heterotrimeric protein that transfers methionyl-initiator tRNA(Met) to the small ribosomal subunit in a ternary complex with GTP. The eIF2 phosphorylated on serine 51 of its alpha subunit [eIF2(alphaP)] acts as competitive inhibitor of its guanine nucleotide exchange factor, eIF2B, impairing formation of the ternary complex and thereby inhibiting translation initiation. eIF2B is comprised of catalytic and regulatory subcomplexes harboring independent eIF2 binding sites; however, it was unknown whether the alpha subunit of eIF2 directly contacts any eIF2B subunits or whether this interaction is modulated by phosphorylation. We found that recombinant eIF2alpha (glutathione S-transferase [GST]-SUI2) bound to the eIF2B regulatory subcomplex in vitro, in a manner stimulated by Ser-51 phosphorylation. Genetic data suggest that this direct interaction also occurred in vivo, allowing overexpressed SUI2 to compete with eIF2(alphaP) holoprotein for binding to the eIF2B regulatory subcomplex. Mutations in SUI2 and in the eIF2B regulatory subunit GCD7 that eliminated inhibition of eIF2B by eIF2(alphaP) also impaired binding of phosphorylated GST-SUI2 to the eIF2B regulatory subunits. These findings provide strong evidence that tight binding of phosphorylated SUI2 to the eIF2B regulatory subcomplex is crucial for the inhibition of eIF2B and attendant downregulation of protein synthesis exerted by eIF2(alphaP). We propose that this regulatory interaction prevents association of the eIF2B catalytic subcomplex with the beta and gamma subunits of eIF2 in the manner required for GDP-GTP exchange. PMID- 11438659 TI - Maf1p, a negative effector of RNA polymerase III in Saccharomyces cerevisiae. AB - Although yeast RNA polymerase III (Pol III) and the auxiliary factors TFIIIC and TFIIIB are well characterized, the mechanisms of class III gene regulation are poorly understood. Previous studies identified MAF1, a gene that affects tRNA suppressor efficiency and interacts genetically with Pol III. We show here that tRNA levels are elevated in maf1 mutant cells. In keeping with the higher levels of tRNA observed in vivo, the in vitro rate of Pol III RNA synthesis is significantly increased in maf1 cell extracts. Mutations in the RPC160 gene encoding the largest subunit of Pol III which reduce tRNA levels were identified as suppressors of the maf1 growth defect. Interestingly, Maf1p is located in the nucleus and coimmunopurifies with epitope-tagged RNA Pol III. These results indicate that Maf1p acts as a negative effector of Pol III synthesis. This potential regulator of Pol III transcription is likely conserved since orthologs of Maf1p are present in other eukaryotes, including humans. PMID- 11438660 TI - A conserved alpha-helical motif mediates the interaction of Sp1-like transcriptional repressors with the corepressor mSin3A. AB - Sp1-like proteins are defined by three highly homologous C(2)H(2) zinc finger motifs that bind GC-rich sequences found in the promoters of a large number of genes essential for mammalian cell homeostasis. Here we report that TIEG2, a transforming growth factor beta-inducible Sp1-like protein with antiproliferative functions, represses transcription through recruitment of the mSin3A-histone deacetylase complex. The interaction of TIEG2 with mSin3A is mediated by an alpha helical repression motif (alpha-HRM) located within the repression domain (R1) of TIEG2. This alpha-HRM specifically associates with the second paired amphipathic helix (PAH2) domain of mSin3A. Mutations in the TIEG2 alpha-HRM domain that disrupt its helical structure abolish its ability to both bind mSin3A and repress transcription. Interestingly, the alpha-HRM is conserved in both the TIEG (TIEG1 and TIEG2) and BTEB (BTEB1, BTEB3, and BTEB4) subfamilies of Sp1-like proteins. The alpha-HRM from these proteins also mediates direct interaction with mSin3A and represses transcription. Surprisingly, we found that the alpha-HRM of the Sp1 like proteins characterized here exhibits structural and functional resemblance to the Sin3A-interacting domain previously described for the basic helix-loop helix protein Mad1. Thus, our study defines a mechanism of transcriptional repression via the interactions of the alpha-HRM with the Sin3-histone deacetylase complex that is utilized by at least five Sp1-like transcriptional factors. More importantly, we demonstrate that a helical repression motif which mediates Sin3 interaction is not an exclusive structural and functional characteristic of the Mad1 subfamily but rather has a wider functional impact on transcriptional repression than previously demonstrated. PMID- 11438661 TI - Mammalian target of rapamycin pathway regulates insulin signaling via subcellular redistribution of insulin receptor substrate 1 and integrates nutritional signals and metabolic signals of insulin. AB - A pathway sensitive to rapamycin, a selective inhibitor of mammalian target of rapamycin (mTOR), down-regulates effects of insulin such as activation of Akt (protein kinase B) via proteasomal degradation of insulin receptor substrate 1 (IRS-1). We report here that the pathway also plays an important role in insulin induced subcellular redistribution of IRS-1 from the low-density microsomes (LDM) to the cytosol. After prolonged insulin stimulation, inhibition of the redistribution of IRS-1 by rapamycin resulted in increased levels of IRS-1 and the associated phosphatidylinositol (PI) 3-kinase in both the LDM and cytosol, whereas the proteasome inhibitor lactacystin increased the levels only in the cytosol. Since rapamycin but not lactacystin enhances insulin-stimulated 2 deoxyglucose (2-DOG) uptake, IRS-1-associated PI 3-kinase localized at the LDM was suggested to be important in the regulation of glucose transport. The amino acid deprivation attenuated and the amino acid excess enhanced insulin-induced Ser/Thr phosphorylation and subcellular redistribution and degradation of IRS-1 in parallel with the effects on phosphorylation of p70 S6 kinase and 4E-BP1. Accordingly, the amino acid deprivation increased and the amino acid excess decreased insulin-stimulated activation of Akt and 2-DOG uptake. Furthermore, 2 DOG uptake was affected by amino acid availability even when the degradation of IRS-1 was inhibited by lactacystin. We propose that subcellular redistribution of IRS-1, regulated by the mTOR-dependent pathway, facilitates proteasomal degradation of IRS-1, thereby down-regulating Akt, and that the pathway also negatively regulates insulin-stimulated glucose transport, probably through the redistribution of IRS-1. This work identifies a novel function of mTOR that integrates nutritional signals and metabolic signals of insulin. PMID- 11438662 TI - Apoptosis triggered by Myc-induced suppression of Bcl-X(L) or Bcl-2 is bypassed during lymphomagenesis. AB - Enforced Bcl-2 expression inhibits Myc-induced apoptosis and cooperates with Myc in transformation. Here we report that the synergy between Bcl-2 and Myc in transforming hematopoietic cells in fact reflects a Myc-induced pathway that selectively suppresses the expression of the Bcl-X(L) or Bcl-2 antiapoptotic protein. Myc activation suppresses Bcl-X(L) RNA and protein levels in cultures of primary myeloid and lymphoid progenitors, and Bcl-X(L) and Bcl-2 expression is inhibited by Myc in precancerous B cells from Emu-myc transgenic mice. The suppression of bcl-X RNA levels by Myc requires de novo protein synthesis, indicating that repression is indirect. Importantly, the suppression of Bcl-2 or Bcl-X(L) by Myc is corrupted during Myc-induced tumorigenesis, as Bcl-2 and/or Bcl-X(L) levels are markedly elevated in over one-half of all lymphomas arising in Emicro-myc transgenic mice. Bcl-2 and/or Bcl-X(L) overexpression did not correlate with loss of ARF or p53 function in tumor cells, indicating that these two apoptotic pathways are inactivated independently. Therefore, the suppression of Bcl-X(L) or Bcl-2 expression represents a physiological Myc-induced apoptotic pathway that is frequently bypassed during lymphomagenesis. PMID- 11438663 TI - Early expressed Clb proteins allow accumulation of mitotic cyclin by inactivating proteolytic machinery during S phase. AB - Periodic accumulation and destruction of mitotic cyclins are important for the initiation and termination of M phase. It is known that both APC(Cdc20) and APC(Hct1) collaborate to destroy mitotic cyclins during M phase. Here we show that this relationship between anaphase-promoting complex (APC) and Clb proteins is reversed in S phase such that the early Clb kinases (Clb3, Clb4, and Clb5 kinases) inactivate APC(Hct1) to allow Clb2 accumulation. This alternating antagonism between APC and Clb proteins during S and M phases constitutes an oscillatory system that generates undulations in the levels of mitotic cyclins. PMID- 11438664 TI - Identification of insulin receptor substrate 1 (IRS-1) and IRS-2 as signaling intermediates in the alpha6beta4 integrin-dependent activation of phosphoinositide 3-OH kinase and promotion of invasion. AB - Expression of the alpha6beta4 integrin increases the invasive potential of carcinoma cells by a mechanism that involves activation of phosphoinositide 3-OH kinase (PI3K). In the present study, we investigated the signaling pathway by which the alpha6beta4 integrin activates PI3K. Neither the alpha6 nor the beta4 cytoplasmic domain contains the consensus binding motif for PI3K, pYMXM, indicating that additional proteins are likely to be involved in the activation of this lipid kinase by the alpha6beta4 integrin. We identified insulin receptor substrate 1 (IRS-1) and IRS-2 as signaling intermediates in the activation of PI3K by the alpha6beta4 integrin. IRS-1 and IRS-2 are cytoplasmic adapter proteins that do not contain intrinsic kinase activity but rather function by recruiting proteins to surface receptors, where they organize signaling complexes. Ligation of the alpha6beta4 receptor promotes tyrosine phosphorylation of IRS-1 and IRS-2 and increases their association with PI3K, as determined by coimmunoprecipitation. Moreover, we identified a tyrosine residue in the cytoplasmic domain of the beta4 subunit, Y1494, that is required for alpha6beta4 dependent phosphorylation of IRS-2 and activation of PI3K in response to receptor ligation. Most importantly, Y1494 is essential for the ability of the alpha6beta4 integrin to promote carcinoma invasion. Taken together, these results imply a key role for the IRS proteins in the alpha6beta4-dependent promotion of carcinoma invasion. PMID- 11438665 TI - Proteolysis of the docking protein HEF1 and implications for focal adhesion dynamics. AB - The dynamic regulation of focal adhesions is implicated in cellular processes of proliferation, differentiation, migration, and apoptosis. The focal adhesion associated docking protein HEF1 is cleaved by caspases during both mitosis and apoptosis. Common to both of these cellular processes is the loss of focal adhesions, transiently during mitosis and permanently during apoptosis. The proteolytic processing of HEF1 during both mitosis and apoptosis therefore posits a general role for HEF1 as a sensor of altered adhesion states. In this study, we find that HEF1 undergoes proteolytic processing specifically in response to cellular detachment, while HEF1 proteolysis is prevented by specific integrin receptor ligation and focal adhesion formation. We show that overexpression of a C-terminal caspase-derived 28-kDa HEF1 peptide causes cellular rounding that is demonstrably separable from apoptosis. Mutation of the divergent helix-loop-helix motif found in 28-kDa HEF1 significantly reduces the induction of apoptosis by this peptide, while deletion of the amino-terminal 28 amino acids of 28-kDa HEF1 completely abrogates the induction of apoptosis. Conversely, these mutations have no effect on the rounding induced by 28-kDa HEF1. Finally, we detect a novel focal adhesion targeting domain located in the C terminus of HEF1 and show that this activity is necessary for HEF1-induced cell spreading. Together, these data suggest that proteolytic and other posttranslational modifications of HEF1 in response to loss of adhesion serve to modulate the disassembly of focal adhesions. PMID- 11438666 TI - The TFIID components human TAF(II)140 and Drosophila BIP2 (TAF(II)155) are novel metazoan homologues of yeast TAF(II)47 containing a histone fold and a PHD finger. AB - The RNA polymerase II transcription factor TFIID comprises the TATA binding protein (TBP) and a set of TBP-associated factors (TAF(II)s). TFIID has been extensively characterized for yeast, Drosophila, and humans, demonstrating a high degree of conservation of both the amino acid sequences of the constituent TAF(II)s and overall molecular organization. In recent years, it has been assumed that all the metazoan TAF(II)s have been identified, yet no metazoan homologues of yeast TAF(II)47 (yTAF(II)47) and yTAF(II)65 are known. Both of these yTAF(II)s contain a histone fold domain (HFD) which selectively heterodimerizes with that of yTAF(II)25. We have cloned a novel mouse protein, TAF(II)140, containing an HFD and a plant homeodomain (PHD) finger, which we demonstrated by immunoprecipitation to be a mammalian TFIID component. TAF(II)140 shows extensive sequence similarity to Drosophila BIP2 (dBIP2) (dTAF(II)155), which we also show to be a component of Drosophila TFIID. These proteins are metazoan homologues of yTAF(II)47 as their HFDs selectively heterodimerize with dTAF(II)24 and human TAF(II)30, metazoan homologues of yTAF(II)25. We further show that yTAF(II)65 shares two domains with the Drosophila Prodos protein, a recently described potential dTAF(II). These conserved domains are critical for yTAF(II)65 function in vivo. Our results therefore identify metazoan homologues of yTAF(II)47 and yTAF(II)65. PMID- 11438667 TI - Smad proteins and hepatocyte growth factor control parallel regulatory pathways that converge on beta1-integrin to promote normal liver development. AB - Smads serve as intracellular mediators of transforming growth factor beta (TGF beta) signaling. After phosphorylation by activated type I TGF-beta receptors, Smad proteins translocate to the nucleus, where they serve as transcription factors and increase or decrease expression of TGF-beta target genes. Mice lacking one copy each of Smad2 and Smad3 suffered midgestation lethality due to liver hypoplasia and anemia, suggesting essential dosage requirements of TGF-beta signal components. This is likely due to abnormal adhesive properties of the mutant hepatocytes, which may result from a decrease in the level of the beta1 integrin and abnormal processing and localization of E-cadherin. Culture of mutant livers in vitro revealed the existence of a parallel developmental pathway mediated by hepatocyte growth factor (HGF), which could rescue the mutant phenotype independent of Smad activation. These pathways merge at the beta1 integrin, the level of which was increased by HGF in the cultured mutant livers. HGF treatment reversed the defects in cell proliferation and hepatic architecture in the Smad2(+/-); Smad3(+/-) livers. PMID- 11438668 TI - Axin facilitates Smad3 activation in the transforming growth factor beta signaling pathway. AB - Axin acts as a negative regulator in Wnt signaling through interaction with various molecules involved in this pathway, including beta-catenin, adenomatous polyposis coli, and glycogen synthase kinase 3beta. We show here that Axin also regulates the effects of Smad3 on the transforming growth factor beta (TGF-beta) signaling pathway. In the absence of activated TGF-beta receptors. Axin physically interacted with Smad3 through its C-terminal region located between the beta-catenin binding site and Dishevelled-homologous domain. An Axin homologue, Axil (also called conductin), also interacted with Smad3. In the absence of ligand stimulation, Axin was colocalized with Smad3 in the cytoplasm in vivo. Upon receptor activation, Smad3 was strongly phosphorylated by TGF-beta type I receptor (TbetaR-I) in the presence of Axin, and dissociated from TbetaR-I and Axin. Moreover, the transcriptional activity of TGF-beta was enhanced by Axin and repressed by an Axin mutant which is able to bind to Smad3. Axin may thus function as an adapter of Smad3, facilitating its activation by TGF-beta receptors for efficient TGF-beta signaling. PMID- 11438669 TI - exo1-Dependent mutator mutations: model system for studying functional interactions in mismatch repair. AB - EXO1 interacts with MSH2 and MLH1 and has been proposed to be a redundant exonuclease that functions in mismatch repair (MMR). To better understand the role of EXO1 in mismatch repair, a genetic screen was performed to identify mutations that increase the mutation rates caused by weak mutator mutations such as exo1Delta and pms1-A130V mutations. In a screen starting with an exo1 mutation, exo1-dependent mutator mutations were obtained in MLH1, PMS1, MSH2, MSH3, POL30 (PCNA), POL32, and RNR1, whereas starting with the weak pms1 allele pms1-A130V, pms1-dependent mutator mutations were identified in MLH1, MSH2, MSH3, MSH6, and EXO1. These mutations only cause weak MMR defects as single mutants but cause strong MMR defects when combined with each other. Most of the mutations obtained caused amino acid substitutions in MLH1 or PMS1, and these clustered in either the ATP-binding region or the MLH1-PMS1 interaction regions of these proteins. The mutations showed two other types of interactions: specific pairs of mutations showed unlinked noncomplementation in diploid strains, and the defect caused by pairs of mutations could be suppressed by high-copy-number expression of a third gene, an effect that showed allele and overexpressed gene specificity. These results support a model in which EXO1 plays a structural role in MMR and stabilizes multiprotein complexes containing a number of MMR proteins. A similar role is proposed for PCNA based on the data presented. PMID- 11438670 TI - Loss of cell cycle checkpoint control in Drosophila Rfc4 mutants. AB - Two alleles of the Drosophila melanogaster Rfc4 (DmRfc4) gene, which encodes subunit 4 of the replication factor C (RFC) complex, cause striking defects in mitotic chromosome cohesion and condensation. These mutations produce larval phenotypes consistent with a role in DNA replication but also result in mitotic chromosomal defects appearing either as premature chromosome condensation-like or precocious sister chromatid separation figures. Though the DmRFC4 protein localizes to all replicating nuclei, it is dispersed from chromatin in mitosis. Thus the mitotic defects appear not to be the result of a direct role for RFC4 in chromosome structure. We also show that the mitotic defects in these two DmRfc4 alleles are the result of aberrant checkpoint control in response to DNA replication inhibition or damage to chromosomes. Not all surveillance function is compromised in these mutants, as the kinetochore attachment checkpoint is operative. Intriguingly, metaphase delay is frequently observed with the more severe of the two alleles, indicating that subsequent chromosome segregation may be inhibited. This is the first demonstration that subunit 4 of RFC functions in checkpoint control in any organism, and our findings additionally emphasize the conserved nature of RFC's involvement in checkpoint control in multicellular eukaryotes. PMID- 11438671 TI - Mitotic phosphorylation prevents the binding of HMGN proteins to chromatin. AB - Condensation of the chromatin fiber and transcriptional inhibition during mitosis is associated with the redistribution of many DNA- and chromatin-binding proteins, including members of the high-mobility-group N (HMGN) family. Here we study the mechanism governing the organization of HMGN proteins in mitosis. Using site-specific antibodies and quantitative gel analysis with proteins extracted from synchronized HeLa cells, we demonstrate that, during mitosis, the conserved serine residues in the nucleosomal binding domain (NBD) of this protein family are highly and specifically phosphorylated. Nucleosome mobility shift assays with both in vitro-phosphorylated proteins and with point mutants bearing negative charges in the NBD demonstrate that the negative charge abolishes the ability of the proteins to bind to nucleosomes. Fluorescence loss of photobleaching demonstrates that, in living cells, the negative charge in the NBD increases the intranuclear mobility of the protein and significantly decreases the relative time that it is bound to chromatin. Expression of wild-type and mutant proteins in HmgN1(-/-) cells indicates that the negatively charged protein is not bound to chromosomes. We conclude that during mitosis the NBD of HMGN proteins is highly phosphorylated and that this modification regulates the interaction of the proteins with chromatin. PMID- 11438672 TI - Conformational switch and role of phosphorylation in PAK activation. AB - p21-activated protein kinases (PAKs) are involved in signal transduction processes initiating a variety of biological responses. They become activated by interaction with Rho-type small GTP-binding proteins Rac and Cdc42 in the GTP bound conformation, thereby relieving the inhibition of the regulatory domain (RD) on the catalytic domain (CD). Here we report on the mechanism of activation and show that proteolytic digestion of PAK produces a heterodimeric RD-CD complex consisting of a regulatory fragment (residues 57 to 200) and a catalytic fragment (residues 201 to 491), which is active in the absence of Cdc42. Cdc42-GppNHp binds with low affinity (K(d) 0.6 microM) to intact kinase, whereas the affinity to the isolated regulatory fragment is much higher (K(d) 18 nM), suggesting that the difference in binding energy is used for the conformational change leading to activation. The full-length kinase, the isolated RD, and surprisingly also their complexes with Cdc42 behave as dimers on a gel filtration column. Cdc42-GppNHp interaction with the RD-CD complex is also of low affinity and does not dissociate the RD from the CD. After autophosphorylation of the kinase domain, Cdc42 binds with high (14 nM) affinity and dissociates the RD-CD complex. Assuming that the RD-CD complex mimics the interaction in native PAK, this indicates that the small G protein may not simply release the RD from the CD. It acts in a more subtle allosteric control mechanism to induce autophosphorylation, which in turn induces the release of the RD and thus full activation. PMID- 11438673 TI - Identification of an overlapping binding domain on Cdc20 for Mad2 and anaphase promoting complex: model for spindle checkpoint regulation. AB - Activation of the anaphase-promoting complex (APC) is required for anaphase initiation and for exit from mitosis in mammalian cells. Cdc20, which specifically recognizes APC substrates involved in the metaphase-to-anaphase transition, plays a pivotal role in APC activation through direct interaction with the APC. The activation of the APC by Cdc20 is prevented by the interaction of Cdc20 with Mad2 when the spindle checkpoint is activated. Using deletion mutagenesis and peptide mapping, we have identified the sequences in Cdc20 that target it to Mad2 and the APC, respectively. These sequences are distinct but overlapping, providing a possible structural explanation for the internal modulation of the APC-Cdc20 complex by Mad2. In the course of these studies, a truncation mutant of Cdc20 (1-153) that constitutively binds Mad2 but fails to bind the APC was identified. Overexpression of this mutant induces the formation of multinucleated cells and increases their susceptibility to undergoing apoptosis when treated with microtubule-inhibiting drugs. Our experiments demonstrate that disruption of the Mad2-Cdc20 interaction perturbs the mitotic checkpoint, leading to premature activation of the APC, sensitizing the cells to the cytotoxic effects of microtubule-inhibiting drugs. PMID- 11438674 TI - Dual interactions of the translational repressor Paip2 with poly(A) binding protein. AB - The cap structure and the poly(A) tail of eukaryotic mRNAs act synergistically to enhance translation. This effect is mediated by a direct interaction of eukaryotic initiation factor 4G and poly(A) binding protein (PABP), which brings about circularization of the mRNA. Of the two recently identified PABP interacting proteins, one, Paip1, stimulates translation, and the other, Paip2, which competes with Paip1 for binding to PABP, represses translation. Here we studied the Paip2-PABP interaction. Biacore data and far-Western analysis revealed that Paip2 contains two binding sites for PABP, one encompassing a 16 amino-acid stretch located in the C terminus and a second encompassing a larger central region. PABP also contains two binding regions for Paip2, one located in the RNA recognition motif (RRM) region and the other in the carboxy-terminal region. A two-to-one stoichiometry for binding of Paip2 to PABP with two independent K(d)s of 0.66 and 74 nM was determined. Thus, our data demonstrate that PABP and Paip2 could form a trimeric complex containing one PABP molecule and two Paip2 molecules. Significantly, only the central Paip2 fragment, which binds with high affinity to the PABP RRM region, inhibits PABP binding to poly(A) RNA and translation. PMID- 11438675 TI - Chk2 activation dependence on Nbs1 after DNA damage. AB - The checkpoint kinase Chk2 has a key role in delaying cell cycle progression in response to DNA damage. Upon activation by low-dose ionizing radiation (IR), which occurs in an ataxia telangiectasia mutated (ATM)-dependent manner, Chk2 can phosphorylate the mitosis-inducing phosphatase Cdc25C on an inhibitory site, blocking entry into mitosis, and p53 on a regulatory site, causing G(1) arrest. Here we show that the ATM-dependent activation of Chk2 by gamma- radiation requires Nbs1, the gene product involved in the Nijmegen breakage syndrome (NBS), a disorder that shares with AT a variety of phenotypic defects including chromosome fragility, radiosensitivity, and radioresistant DNA synthesis. Thus, whereas in normal cells Chk2 undergoes a time-dependent increased phosphorylation and induction of catalytic activity against Cdc25C, in NBS cells null for Nbs1 protein, Chk2 phosphorylation and activation are both defective. Importantly, these defects in NBS cells can be complemented by reintroduction of wild-type Nbs1, but neither by a carboxy-terminal deletion mutant of Nbs1 at amino acid 590, unable to form a complex with and to transport Mre11 and Rad50 in the nucleus, nor by an Nbs1 mutated at Ser343 (S343A), the ATM phosphorylation site. Chk2 nuclear expression is unaffected in NBS cells, hence excluding a mislocalization as the cause of failed Chk2 activation in Nbs1-null cells. Interestingly, the impaired Chk2 function in NBS cells correlates with the inability, unlike normal cells, to stop entry into mitosis immediately after irradiation, a checkpoint abnormality that can be corrected by introduction of the wild-type but not the S343A mutant form of Nbs1. Altogether, these findings underscore the crucial role of a functional Nbs1 complex in Chk2 activation and suggest that checkpoint defects in NBS cells may result from the inability to activate Chk2. PMID- 11438676 TI - The novel transcription factor e(y)2 interacts with TAF(II)40 and potentiates transcription activation on chromatin templates. AB - Weak hypomorph mutations in the enhancer of yellow genes, e(y)1 and e(y)2, of Drosophila melanogaster were discovered during the search for genes involved in the organization of interaction between enhancers and promoters. Previously, the e(y)1 gene was cloned and found to encode TAF(II)40 protein. Here we cloned the e(y)2 gene and demonstrated that it encoded a new ubiquitous evolutionarily conserved transcription factor. The e(y)2 gene is located at 10C3 (36.67) region and is expressed at all stages of Drosophila development. It encodes a 101-amino acid protein, e(y)2. Vertebrates, insects, protozoa, and plants have proteins which demonstrate a high degree of homology to e(y)2. The e(y)2 protein is localized exclusively to the nuclei and is associated with numerous sites along the entire length of the salivary gland polytene chromosomes. Both genetic and biochemical experiments demonstrate an interaction between e(y)2 and TAF(II)40, while immunoprecipitation studies demonstrate that the major complex, including both proteins, appears to be distinct from TFIID. Furthermore, we provide genetic evidence suggesting that the carboxy terminus of dTAF(II)40 is important for mediating this interaction. Finally, using an in vitro transcription system, we demonstrate that recombinant e(y)2 is able to enhance transactivation by GAL4 VP16 on chromatin but not on naked DNA templates, suggesting that this novel protein is involved in the regulation of transcription. PMID- 11438677 TI - Recognition of RNA branch point sequences by the KH domain of splicing factor 1 (mammalian branch point binding protein) in a splicing factor complex. AB - Mammalian splicing factor 1 (SF1; also mammalian branch point binding protein [mBBP]; hereafter SF1/mBBP) specifically recognizes the seven-nucleotide branch point sequence (BPS) located at 3' splice sites and participates in the assembly of early spliceosomal complexes. SF1/mBBP utilizes a "maxi-K homology" (maxi-KH) domain for recognition of the single-stranded BPS and requires a cooperative interaction with splicing factor U2AF65 bound to an adjacent polypyrimidine tract (PPT) for high-affinity binding. To investigate how the KH domain of SF1/mBBP recognizes the BPS in conjunction with U2AF and possibly other proteins, we constructed a transcriptional reporter system utilizing human immunodeficiency virus type 1 Tat fusion proteins and examined the RNA-binding specificity of the complex using KH domain and RNA-binding site mutants. We first established that SF1/mBBP and U2AF cooperatively assemble in our reporter system at RNA sites composed of the BPS, PPT, and AG dinucleotide found at 3' splice sites, with endogenous proteins assembled along with the Tat fusions. We next found that the activities of the Tat fusion proteins on different BPS variants correlated well with the known splicing efficiencies of the variants, supporting a model in which the SF1/mBBP-BPS interaction helps determine splicing efficiency prior to the U2 snRNP-BPS interaction. Finally, the likely RNA-binding surface of the maxi-KH domain was identified by mutagenesis and appears similar to that used by "simple" KH domains, involving residues from two putative alpha helices, a highly conserved loop, and parts of a beta sheet. Using a homology model constructed from the cocrystal structure of a Nova KH domain-RNA complex (Lewis et al., Cell 100:323-332, 2000), we propose a plausible arrangement for SF1/mBBP-U2AF complexes assembled at 3' splice sites. PMID- 11438678 TI - Drosophila Med6 is required for elevated expression of a large but distinct set of developmentally regulated genes. AB - Mediator is the evolutionarily conserved coactivator required for the integration and recruitment of diverse regulatory signals to basal transcription machinery. To elucidate the functions of metazoan Mediator, we isolated Drosophila melanogaster Med6 mutants. dMed6 is essential for viability and/or proliferation of most cells. dMed6 mutants failed to pupate and died in the third larval instar with severe proliferation defects in imaginal discs and other larval mitotic cells. cDNA microarray, quantitative reverse transcription-PCR, and in situ expression analyses of developmentally regulated genes in dMed6 mutants showed that transcriptional activation of many, but not all, genes was affected. Among the genes found to be affected were some that play a role in cell proliferation and metabolism. Therefore, dMed6 is required in most cells for transcriptional regulation of many genes important for diverse aspects of Drosophila development. PMID- 11438679 TI - Impaired abdominal wall development and deficient wound healing in mice lacking aortic carboxypeptidase-like protein. AB - Aortic carboxypeptidase-like protein (ACLP) is a member of a diverse group of proteins that contain a domain with similarity to that of the Dictyostelium discoideum protein discoidin I. The discoidin domain has been identified in mammalian milk fat globule membrane proteins, blood coagulation factors, and receptor tyrosine kinases, where it may facilitate cell aggregation, adhesion, or cell-cell recognition. Here we show that ACLP is a secreted protein that associates with the extracellular matrix (ECM). During mouse embryogenesis, ACLP is abundantly expressed in the ECM of collagen-rich tissues, including the vasculature, dermis, and the developing skeleton. We deleted the ACLP gene in mice by homologous recombination. The majority of ACLP(-/-) mice die perinatally due to gastroschisis, a severe disruption of the anterior abdominal wall and herniation of the abdominal organs. ACLP(-/-) mice that survived to adulthood developed nonhealing skin wounds. Following injury by a dermal punch biopsy, ACLP(-/-) mice exhibited deficient wound healing compared with controls. In addition, dermal fibroblasts isolated from ACLP(-/-) 18.5-day-postconception embryos exhibited a reduced proliferative capacity compared with wild-type cells. These results indicate that ACLP is an ECM protein that is essential for embryonic development and dermal wound healing processes. PMID- 11438680 TI - G(alpha)11 signaling through ARF6 regulates F-actin mobilization and GLUT4 glucose transporter translocation to the plasma membrane. AB - The action of insulin to recruit the intracellular GLUT4 glucose transporter to the plasma membrane of 3T3-L1 adipocytes is mimicked by endothelin 1, which signals through trimeric G(alpha)q or G(alpha)11 proteins. Here we report that murine G(alpha)11 is most abundant in fat and that expression of the constitutively active form of G(alpha)11 [G(alpha)11(Q209L)] in 3T3-L1 adipocytes causes recruitment of GLUT4 to the plasma membrane and stimulation of 2 deoxyglucose uptake. In contrast to the action of insulin on GLUT4, the effects of endothelin 1 and G(alpha)11 were not inhibited by the phosphatidylinositol 3 kinase inhibitor wortmannin at 100 nM. Signaling by insulin, endothelin 1, or G(alpha)11(Q209L) also mobilized cortical F-actin in cultured adipocytes. Importantly, GLUT4 translocation caused by all three agents was blocked upon disassembly of F-actin by latrunculin B, suggesting that the F-actin polymerization caused by these agents may be required for their effects on GLUT4. Remarkably, expression of a dominant inhibitory form of the actin-regulatory GTPase ARF6 [ARF6(T27N)] in cultured adipocytes selectively inhibited both F actin formation and GLUT4 translocation in response to endothelin 1 but not insulin. These data indicate that ARF6 is a required downstream element in endothelin 1 signaling through G(alpha)11 to regulate cortical actin and GLUT4 translocation in cultured adipocytes, while insulin action involves different signaling pathways. PMID- 11438681 TI - ADP-ribosylation factor 6 delineates separate pathways used by endothelin 1 and insulin for stimulating glucose uptake in 3T3-L1 adipocytes. AB - In 3T3-L1 adipocytes, both insulin and endothelin 1 stimulate glucose transport via translocation of the GLUT4 glucose carrier from an intracellular compartment to the cell surface. Yet it remains uncertain as to whether both hormones utilize identical pathways and to what extent each depends on the heterotrimeric G protein Galphaq as an intermediary signaling molecule. In this study, we used a novel inducible system to rapidly and synchronously activate expression of a dominant inhibitory form of ADP-ribosylation factor 6, ARF6(T27N), in 3T3-L1 adipocytes and assessed its effects on insulin- and endothelin-stimulated hexose uptake. Expression of ARF6(T27N) in 3T3-L1 adipocytes was without effect on the ability of insulin to stimulate either 2-deoxyglucose uptake or the translocation of GLUT4 or GLUT1 to the plasma membrane. However, the same ARF6 inhibitory mutant blocked the stimulation of hexose uptake and GLUT4 translocation in response to either endothelin 1 or an activated form of Galphaq, Galphaq(Q209L). These results suggest that endothelin stimulates glucose transport through a pathway that is distinct from that utilized by insulin but is likely to depend on both a heterotrimeric G protein from the Gq family and the small G protein ARF6. These data are consistent with the interpretation that endothelin and insulin stimulate functionally different pools of glucose transporters to be redistributed to the plasma membrane. PMID- 11438682 TI - The human formin-binding protein 17 (FBP17) interacts with sorting nexin, SNX2, and is an MLL-fusion partner in acute myelogeneous leukemia. AB - We have cloned a fusion partner of the MLL gene at 11q23 and identified it as the gene encoding the human formin-binding protein 17, FBP17. It maps to chromosome 9q34 centromeric to ABL. The gene fusion results from a complex chromosome rearrangement that was resolved by fluorescence in situ hybridization with various probes on chromosomes 9 and 11 as an ins(11;9)(q23;q34)inv(11)(q13q23). The rearrangement resulted in a 5'-MLL/FBP17-3' fusion mRNA. We retrovirally transduced murine-myeloid progenitor cells with MLL/FBP17 to test its transforming ability. In contrast to MLL/ENL, MLL/ELL and other MLL-fusion genes, MLL/FBP17 did not give a positive readout in a serial replating assay. Therefore, we assume that additional cooperating genetic abnormalities might be needed to establish a full malignant phenotype. FBP17 consists of a C-terminal Src homology 3 domain and an N-terminal region that is homologous to the cell division cycle protein, cdc15, a regulator of the actin cytoskeleton in Schizosaccharomyces pombe. Both domains are separated by a consensus Rho-binding motif that has been identified in different Rho-interaction partners such as Rhotekin and Rhophilin. We evaluated whether FBP17 and members of the Rho family interact in vivo with a yeast two-hybrid assay. None of the various Rho proteins tested, however, interacted with FBP17. We screened a human kidney library and identified a sorting nexin, SNX2, as a protein interaction partner of FBP17. These data provide a link between the epidermal growth factor receptor pathway and an MLL fusion protein. PMID- 11438683 TI - BeF(3)(-) acts as a phosphate analog in proteins phosphorylated on aspartate: structure of a BeF(3)(-) complex with phosphoserine phosphatase. AB - Protein phosphoaspartate bonds play a variety of roles. In response regulator proteins of two-component signal transduction systems, phosphorylation of an aspartate residue is coupled to a change from an inactive to an active conformation. In phosphatases and mutases of the haloacid dehalogenase (HAD) superfamily, phosphoaspartate serves as an intermediate in phosphotransfer reactions, and in P-type ATPases, also members of the HAD family, it serves in the conversion of chemical energy to ion gradients. In each case, lability of the phosphoaspartate linkage has hampered a detailed study of the phosphorylated form. For response regulators, this difficulty was recently overcome with a phosphate analog, BeF(3)(-), which yields persistent complexes with the active site aspartate of their receiver domains. We now extend the application of this analog to a HAD superfamily member by solving at 1.5-A resolution the x-ray crystal structure of the complex of BeF(3)(-) with phosphoserine phosphatase (PSP) from Methanococcus jannaschii. The structure is comparable to that of a phosphoenzyme intermediate: BeF(3)(-) is bound to Asp-11 with the tetrahedral geometry of a phosphoryl group, is coordinated to Mg(2+), and is bound to residues surrounding the active site that are conserved in the HAD superfamily. Comparison of the active sites of BeF(3)(-) x PSP and BeF(3)(-) x CeY, a receiver domain/response regulator, reveals striking similarities that provide insights into the function not only of PSP but also of P-type ATPases. Our results indicate that use of BeF(3)(-) for structural studies of proteins that form phosphoaspartate linkages will extend well beyond response regulators. PMID- 11438684 TI - Deciphering the design of the tropomyosin molecule. AB - The crystal structure at 2.0-A resolution of an 81-residue N-terminal fragment of muscle alpha-tropomyosin reveals a parallel two-stranded alpha-helical coiled coil structure with a remarkable core. The high alanine content of the molecule is clustered into short regions where the local 2-fold symmetry is broken by a small (approximately 1.2-A) axial staggering of the helices. The joining of these regions with neighboring segments, where the helices are in axial register, gives rise to specific bends in the molecular axis. We observe such bends to be widely distributed in two-stranded alpha-helical coiled-coil proteins. This asymmetric design in a dimer of identical (or highly similar) sequences allows the tropomyosin molecule to adopt multiple bent conformations. The seven alanine clusters in the core of the complete molecule (which spans seven monomers of the actin helix) promote the semiflexible winding of the tropomyosin filament necessary for its regulatory role in muscle contraction. PMID- 11438685 TI - Long-term plasticity in interneurons of the dentate gyrus. AB - Single interneurons influence thousands of postsynaptic principal cells, and the control of interneuronal excitability is an important regulator of the computational properties of the hippocampus. However, the mechanisms underlying long-term alterations in the input-output functions of interneurons are not fully understood. We report a mechanism of interneuronal plasticity that leads to the functional enhancement of the gain of glutamatergic inputs in the absence of long term potentiation of the excitatory synaptic currents. Interneurons in the dentate gyrus exhibit a characteristic, limited (approximately 8 mV) depolarization of their resting membrane potential after high-frequency stimulation of the perforant path. The depolarization can be observed with either whole-cell or perforated patch electrodes, and it lasts in excess of 3 h. The long-term depolarization is specific to interneurons, because granule cells do not show it. The depolarization requires the activation of Ca(2+)-permeable alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors and the rise of intracellular Ca(2+), but not N-methyl-d-aspartate (NMDA) receptor activation. Data on the maintenance of the depolarization point to a major role for a long term change in the rate of electrogenic Na(+)/K(+)-ATPase pump function in interneurons. As a result of the depolarization, interneurons after the tetanus respond with action potential discharges to previously subthreshold excitatory postsynaptic potentials (EPSPs), even though the EPSPs are not potentiated. These results demonstrate that the plastic nature of the interneuronal resting membrane potential underlies a unique form of long-term regulation of the gain of excitatory inputs to gamma-aminobutyric acid (GABA)ergic neurons. PMID- 11438686 TI - The highly recombinogenic bz locus lies in an unusually gene-rich region of the maize genome. AB - The bronze (bz) locus exhibits the highest rate of recombination of any gene in higher plants. To investigate the possible basis of this high rate of recombination, we have analyzed the physical organization of the region around the bz locus. Two adjacent bacterial artificial chromosome clones, comprising a 240-kb contig centered around the Bz-McC allele, were isolated, and 60 kb of contiguous DNA spanning the two bacterial artificial chromosome clones was sequenced. We find that the bz locus lies in an unusually gene-rich region of the maize genome. Ten genes, at least eight of which are shown to be transcribed, are contained in a 32-kb stretch of DNA that is uninterrupted by retrotransposons. We have isolated nearly full length cDNAs corresponding to the five proximal genes in the cluster. The average intertranscript distance between them is just 1 kb, revealing a surprisingly compact packaging of adjacent genes in this part of the genome. At least 11 small insertions, including several previously described miniature inverted repeat transposable elements, were detected in the introns and 3' untranslated regions of genes and between genes. The gene-rich region is flanked at the proximal and distal ends by retrotransposon blocks. Thus, the maize genome appears to have scattered regions of high gene density similar to those found in other plants. The unusually high rate of intragenic recombination seen in bz may be related to the very high gene density of the region. PMID- 11438687 TI - Antagonistic forces generated by myosin II and cytoplasmic dynein regulate microtubule turnover, movement, and organization in interphase cells. AB - Photoactivation of caged fluorescent tubulin was used mark the microtubule (MT) lattice and monitor MT behavior in interphase cells. A broadening of the photoactivated region occurred as MTs moved bidirectionally. MT movement was not inhibited when MT assembly was suppressed with nocodazole or Taxol; MT movement was suppressed by inhibition of myosin light chain kinase with ML7 or by a peptide inhibitor. Conversely, MT movement was increased after inhibition of cytoplasmic dynein with the antibody 70.1. In addition, the half-time for MT turnover was decreased in cells treated with ML7. These results demonstrate that myosin II and cytoplasmic dynein contribute to a balance of forces that regulates MT organization, movement, and turnover in interphase cells. PMID- 11438688 TI - Biosynthesis of the thiazole moiety of thiamin in Escherichia coli: identification of an acyldisulfide-linked protein--protein conjugate that is functionally analogous to the ubiquitin/E1 complex. AB - A covalently linked protein--protein conjugate between ThiF and ThiS thiocarboxylate was found in a partially purified coexpressed ThiF/ThiS protein mixture by using Fourier transform mass spectrometry. The Cys-184 of ThiF and the C terminus of ThiS thiocarboxylate were identified to be involved in the formation of this complex by using both mutagenesis and chemical modification methods. A complementation study of Escherichia coli thiF(-) using thiF(C184S) suggests that this conjugate is an essential intermediate involved in the biosynthesis of the thiazole moiety of thiamin. This ThiF/ThiS conjugate is the first characterized example of a unique acyldisulfide intermediate in a biosynthetic system. This protein conjugate is also an example of an ubiquitin-E1 like protein-protein conjugate in prokaryotes and supports a strong evolutionary link between thiamin biosynthesis and the ubiquitin conjugating system. PMID- 11438689 TI - In vitro generated antibodies specific for telomeric guanine-quadruplex DNA react with Stylonychia lemnae macronuclei. AB - Most eukaryotic telomeres contain a repeating motif with stretches of guanine residues that form a 3'-terminal overhang extending beyond the telomeric duplex region. The telomeric repeat of hypotrichous ciliates, d(T(4)G(4)), forms a 16 nucleotide 3'-overhang. Such sequences can adopt parallel-stranded as well as antiparallel-stranded quadruplex conformations in vitro. Although it has been proposed that guanine-quadruplex conformations may have important cellular roles including telomere function, recombination, and transcription, evidence for the existence of this DNA structure in vivo has been elusive to date. We have generated high-affinity single-chain antibody fragment (scFv) probes for the guanine-quadruplex formed by the Stylonychia telomeric repeat, by ribosome display from the Human Combinatorial Antibody Library. Of the scFvs selected, one (Sty3) had an affinity of K(d) = 125 pM for the parallel-stranded guanine quadruplex and could discriminate with at least 1,000-fold specificity between parallel or antiparallel quadruplex conformations formed by the same sequence motif. A second scFv (Sty49) bound both the parallel and antiparallel quadruplex with similar (K(d) = 3--5 nM) affinity. Indirect immunofluorescence studies show that Sty49 reacts specifically with the macronucleus but not the micronucleus of Stylonychia lemnae. The replication band, the region where replication and telomere elongation take place, was also not stained, suggesting that the guanine quadruplex is resolved during replication. Our results provide experimental evidence that the telomeres of Stylonychia macronuclei adopt in vivo a guanine quadruplex structure, indicating that this structure may have an important role for telomere functioning. PMID- 11438690 TI - Protacs: chimeric molecules that target proteins to the Skp1-Cullin-F box complex for ubiquitination and degradation. AB - The intracellular levels of many proteins are regulated by ubiquitin-dependent proteolysis. One of the best-characterized enzymes that catalyzes the attachment of ubiquitin to proteins is a ubiquitin ligase complex, Skp1-Cullin-F box complex containing Hrt1 (SCF). We sought to artificially target a protein to the SCF complex for ubiquitination and degradation. To this end, we tested methionine aminopeptidase-2 (MetAP-2), which covalently binds the angiogenesis inhibitor ovalicin. A chimeric compound, protein-targeting chimeric molecule 1 (Protac-1), was synthesized to recruit MetAP-2 to SCF. One domain of Protac-1 contains the I kappa B alpha phosphopeptide that is recognized by the F-box protein beta-TRCP, whereas the other domain is composed of ovalicin. We show that MetAP-2 can be tethered to SCF(beta-TRCP), ubiquitinated, and degraded in a Protac-1-dependent manner. In the future, this approach may be useful for conditional inactivation of proteins, and for targeting disease-causing proteins for destruction. PMID- 11438691 TI - Altered potassium balance and aldosterone secretion in a mouse model of human congenital long QT syndrome. AB - The voltage-dependent K(+) channel responsible for the slowly activating delayed K(+) current I(Ks) is composed of pore-forming KCNQ1 and regulatory KCNE1 subunits, which are mutated in familial forms of cardiac long QT syndrome. Because KCNQ1 and KCNE1 genes also are expressed in epithelial tissues, such as the kidneys and the intestine, we have investigated the adaptation of KCNE1 deficient mice to different K(+) and Na(+) intakes. On a normal K(+) diet, homozygous kcne1(-/-) mice exhibit signs of chronic volume depletion associated with fecal Na(+) and K(+) wasting and have lower plasma K(+) concentration and higher levels of aldosterone than wild-type mice. Although plasma aldosterone can be suppressed by low K(+) diets or stimulated by low Na(+) diets, a high K(+) diet provokes a tremendous increase of plasma aldosterone levels in kcne1(-/-) mice as compared with wild-type mice (7.1-fold vs. 1.8-fold) despite lower plasma K(+) in kcne1(-/-) mice. This exacerbated aldosterone production in kcne1(-/-) mice is accompanied by an abnormally high plasma renin concentration, which could partly explain the hyperaldosteronism. In addition, we found that KCNE1 and KCNQ1 mRNAs are expressed in the zona glomerulosa of adrenal glands where I(Ks) may directly participate in the control of aldosterone production by plasma K(+). These results, which show that KCNE1 and I(Ks) are involved in K(+) homeostasis, might have important implications for patients with I(Ks)-related long QT syndrome, because hypokalemia is a well known risk factor for the occurrence of torsades de pointes ventricular arrhythmia. PMID- 11438692 TI - Cloning and functional analysis of two gibberellin 3 beta -hydroxylase genes that are differently expressed during the growth of rice. AB - We have cloned two gibberellin (GA) 3 beta-hydroxylase genes, OsGA3ox1 and OsGA3ox2, from rice by screening a genomic library with a DNA fragment obtained by PCR using degenerate primers. We have used full-scan GC-MS and Kovats retention indices to show function for the two encoded recombinant fusion proteins. Both proteins show 3 beta-hydroxylase activity for the steps GA(20) to GA(1), GA(5) to GA(3), GA(44) to GA(38), and GA(9) to GA(4). In addition, indirect evidence suggests that the OsGA3ox1 protein also has 2,3-desaturase activity, which catalyzes the steps GA(9) to 2,3-dehydro-GA(9) and GA(20) to GA(5) (2,3-dehydro GA(20)), and 2 beta-hydroxylase activity, which catalyzes the steps GA(1) to GA(8) and GA(4) to GA(34). Molecular and linkage analysis maps the OsGA3ox1 gene to the distal end of the short arm of chromosome 5; the OsGA3ox2 gene maps to the distal end of the short arm of chromosome 1 that corresponds to the D18 locus. The association of the OsGA3ox2 gene with the d18 locus is confirmed by sequence and complementation analysis of three d18 alleles. Complementation of the d18-AD allele with the OxGA3ox2 gene results in transgenic plants with a normal phenotype. Although both genes show transient expression, the highest level for OsGA3ox1 is from unopened flower. The highest level for OsGA3ox2 is from elongating leaves. PMID- 11438693 TI - Genome-wide gene expression profiles of the developing mouse hippocampus. AB - We have analyzed the developmental molecular programs of the mouse hippocampus, a cortical structure critical for learning and memory, by means of large-scale DNA microarray techniques. Of 11,000 genes and expressed sequence tags examined, 1,926 showed dynamic changes during hippocampal development from embryonic day 16 to postnatal day 30. Gene-cluster analysis was used to group these genes into 16 distinct clusters with striking patterns that appear to correlate with major developmental hallmarks and cellular events. These include genes involved in neuronal proliferation, differentiation, and synapse formation. A complete list of the transcriptional changes has been compiled into a comprehensive gene profile database (http://BrainGenomics.Princeton.edu), which should prove valuable in advancing our understanding of the molecular and genetic programs underlying both the development and the functions of the mammalian brain. PMID- 11438694 TI - A germ-line Tsc1 mutation causes tumor development and embryonic lethality that are similar, but not identical to, those caused by Tsc2 mutation in mice. AB - Tuberous sclerosis (TS) is characterized by the development of hamartomas in various organs and is caused by a germ-line mutation in either TSC1 or TSC2 tumor suppressor genes. From the symptomatic resemblance among TS patients, involvement of TSC1 and TSC2 products in a common pathway has been suggested. Here, to analyze the function of the Tsc1 product, we established a line of Tsc1 (TSC1 homologue) knockout mouse by gene targeting. Heterozygous Tsc1 mutant (Tsc1(+/-)) mice developed renal and extra-renal tumors such as hepatic hemangiomas. In these tumors, loss of wild-type Tsc1 allele was observed. Homozygous Tsc1 mutants died around embryonic days 10.5-11.5, frequently associated with neural tube unclosure. As a whole, phenotypes of Tsc1 knockout mice resembled those of Tsc2 knockout mice previously reported, suggesting that the presumptive common pathway for Tsc1 and Tsc2 products may also exist in mice. Notably, however, development of renal tumors in Tsc1(+/-) mice was apparently slower than that in Tsc2(+/-) mice. The Tsc1 knockout mouse described here will be a useful model to elucidate the function of Tsc1 and Tsc2 products as well as pathogenesis of TS. PMID- 11438695 TI - Location and properties of metal-binding sites on the human prion protein. AB - Although a functional role in copper binding has been suggested for the prion protein, evidence for binding at affinities characteristic of authentic metal binding proteins has been lacking. By presentation of copper(II) ions in the presence of the weak chelator glycine, we have now characterized two high affinity binding sites for divalent transition metals within the human prion protein. One is in the N-terminal octapeptide-repeat segment and has a K(d) for copper(II) of 10(-14) M, with other metals (Ni(2+), Zn(2+), and Mn(2+)) binding three or more orders of magnitude more weakly. However, NMR and fluorescence data reveal a previously unreported second site around histidines 96 and 111, a region of the molecule known to be crucial for prion propagation. The K(d) for copper(II) at this site is 4 x 10(-14) M, whereas nickel(II), zinc(II), and manganese(II) bind 6, 7, and 10 orders of magnitude more weakly, respectively, regardless of whether the protein is in its oxidized alpha-helical (alpha-PrP) or reduced beta-sheet (beta-PrP) conformation. A role for prion protein (PrP) in copper metabolism or transport seems likely and disturbance of this function may be involved in prion-related neurotoxicity. PMID- 11438696 TI - Regulatory function of in vivo anergized CD4(+) T cells. AB - It has been suggested that anergic T cells may not be only inert cells but may rather play an active role, for example by regulating immune responses. We have previously reported the existence of "anergic" IL-10-producing CD4(+) T cells generated in vivo by continuous antigenic stimulation. Using a gene transfer system where the antigen recognized by such T cells is expressed in skeletal muscle by two different DNA viral vectors, we show that these cells not only remain tolerant toward their cognate antigen but also can suppress the immune response of naive T cells against the immunogenic adenoviral proteins. Furthermore, they can completely inhibit tissue destruction that takes place as a result of an immune response. The system presented here is unique in that the T cells have been anergized in vivo, their antigen specificity and functional status are known, and the amount, form, and timing of antigen expression can be manipulated. This model will therefore permit us to carefully dissect the mechanisms by which these anergic T cells regulate the priming and/or effector function of naive T cells. PMID- 11438697 TI - A role for muscarinic excitation: control of specific singing behavior by activation of the adenylate cyclase pathway in the brain of grasshoppers. AB - Muscarinic acetylcholine receptors exert slow and prolonged synaptic effects in both vertebrate and invertebrate nervous systems. Through activation of G proteins, they typically decrease intracellular cAMP levels by inhibition of adenylate cyclase or stimulate phospholipase C and the turnover of inositol phosphates. In insects, muscarinic receptors have been credited with two main functions: inhibition of transmitter release from sensory neuron terminals and regulation of the excitability of motoneurons and interneurons. Our pharmacological studies with intact and behaving grasshoppers revealed a functional role for muscarinic acetylcholine receptors as being the basis for specific arousal in defined areas of the brain, underlying the selection and control of acoustic communication behavior. Periodic injections of acetylcholine into distinct areas of the brain elicited songs of progressively increasing duration. Coinjections of the muscarinic receptor antagonist scopolamine and periodic stimulations with muscarine identified muscarinic receptor activation as being the basis for the underlying accumulation of excitation. In contrast to reports from other studies on functional circuits, muscarinic excitation was apparently mediated by activation of the adenylate cyclase pathway. Stimulation of adenylate cyclase with forskolin and of protein kinase A with 8-Br-cAMP mimicked the stimulatory effects of muscarine whereas inhibition of adenylate cyclase with SQ22536 and of protein kinase A with H-89 and Rp-cAMPs suppressed muscarine-stimulated singing behavior. Activation of adenylate cyclase by muscarinic receptors has previously been reported from studies on membrane preparations and heterologous expression systems, but a physiological significance of this pathway remained to be demonstrated in an in vivo preparation. PMID- 11438698 TI - Dual control of LIF expression and LIF receptor function regulate Stat3 activation at the onset of uterine receptivity and embryo implantation. AB - Leukemia inhibitory factor (LIF) expression in the uterus is essential for embryo implantation in mice. Here we describe the spatial and temporal regulation of LIF signaling in vivo by using tissues isolated from uteri on different days over the implantation period. During this time, LIF receptors are expressed predominantly in the luminal epithelium (LE) of the uterus. Isolated epithelium responds to LIF by phosphorylation and nuclear translocation of signal transducer and activator of transcription (Stat) 3, but not by an increase in mitogen-activated protein kinase levels. The related cytokines Il-6, ciliary neurotrophic factor, as well as epidermal growth factor, do not activate Stat3, although epidermal growth factor stimulates mitogen-activated protein kinase. In vivo Stat3 activation is induced by LIF alone, resulting in the localization of Stat3 specifically to the nuclei of the LE coinciding with the onset of uterine receptivity. The responsiveness of the LE to LIF is regulated temporally, with Stat activation being restricted to day 4 of pregnancy despite the presence of constant levels of LIF receptor throughout the preimplantation period. Uterine receptivity is therefore under dual control and is regulated by both the onset of LIF expression in the endometrial glands and the release from inhibition of receptor function in the LE. PMID- 11438699 TI - A highly conserved protein family interacting with the fragile X mental retardation protein (FMRP) and displaying selective interactions with FMRP related proteins FXR1P and FXR2P. AB - The absence of the fragile X mental retardation protein (FMRP), encoded by the FMR1 gene, is responsible for pathologic manifestations in the Fragile X Syndrome, the most frequent cause of inherited mental retardation. FMRP is an RNA binding protein associated with polysomes as part of a messenger ribonucleoprotein (mRNP) complex. Although its function is poorly understood, various observations suggest a role in local protein translation at neuronal dendrites and in dendritic spine maturation. We present here the identification of CYFIP1/2 (Cytoplasmic FMRP Interacting Proteins) as FMRP interactors. CYFIP1/2 share 88% amino acid sequence identity and represent the two members in humans of a highly conserved protein family. Remarkably, whereas CYFIP2 also interacts with the FMRP-related proteins FXR1P/2P, CYFIP1 interacts exclusively with FMRP. FMRP- CYFIP interaction involves the domain of FMRP also mediating homo- and heteromerization, thus suggesting a competition between interaction among the FXR proteins and interaction with CYFIP. CYFIP1/2 are proteins of unknown function, but CYFIP1 has recently been shown to interact with the small GTPase Rac1, which is implicated in development and maintenance of neuronal structures. Consistent with FMRP and Rac1 localization in dendritic fine structures, CYFIP1/2 are present in synaptosomal extracts. PMID- 11438700 TI - Activation of NF-kappa B by nontypeable Hemophilus influenzae is mediated by toll like receptor 2-TAK1-dependent NIK-IKK alpha /beta-I kappa B alpha and MKK3/6-p38 MAP kinase signaling pathways in epithelial cells. AB - Nontypeable Hemophilus influenzae (NTHi) is an important human pathogen in both children and adults. In children, it causes otitis media, the most common childhood infection and the leading cause of conductive hearing loss in the United States. In adults, it causes lower respiratory tract infections in the setting of chronic obstructive pulmonary disease, the fourth leading cause of death in the United States. The molecular mechanisms underlying the pathogenesis of NTHi-induced infections remain undefined, but they may involve activation of NF-kappa B, a transcriptional activator of multiple host defense genes involved in immune and inflammatory responses. Here, we show that NTHi strongly activates NF-kappa B in human epithelial cells via two distinct signaling pathways, NF kappa B translocation-dependent and -independent pathways. The NF-kappa B translocation-dependent pathway involves activation of NF-kappa B inducing kinase (NIK)--IKK alpha/beta complex leading to I kappa B alpha phosphorylation and degradation, whereas the NF-kappa B translocation-independent pathway involves activation of MKK3/6--p38 mitogen-activated protein (MAP) kinase pathway. Bifurcation of NTHi-induced NIK-IKK alpha/beta-I kappa B alpha and MKK3/6--p38 MAP kinase pathways may occur at transforming growth factor-beta activated kinase 1 (TAK1). Furthermore, we show that toll-like receptor 2 (TLR2) is required for NTHi-induced NF-kappa B activation. In addition, several key inflammatory mediators including IL-1 beta, IL-8, and tumor necrosis factor-alpha are up regulated by NTHi. Finally, P6, a 16-kDa lipoprotein highly conserved in the outer membrane of all NTHi and H. influenzae type b strains, appears to also activate NF-kappa B via similar signaling pathways. Taken together, our results demonstrate that NTHi activates NF-kappa B via TLR2-TAK1-dependent NIK--IKK alpha/beta-I kappa B alpha and MKK3/6--p38 MAP kinase signaling pathways. These studies may bring new insights into molecular pathogenesis of NTHi-induced infections and open up new therapeutic targets for these diseases. PMID- 11438701 TI - Subnuclear targeting of Runx/Cbfa/AML factors is essential for tissue-specific differentiation during embryonic development. AB - Runx (Cbfa/AML) transcription factors are critical for tissue-specific gene expression. A unique targeting signal in the C terminus directs Runx factors to discrete foci within the nucleus. Using Runx2/CBFA1/AML3 and its essential role in osteogenesis as a model, we investigated the fundamental importance of fidelity of subnuclear localization for tissue differentiating activity by deleting the intranuclear targeting signal via homologous recombination. Mice homozygous for the deletion (Runx2 Delta C) do not form bone due to maturational arrest of osteoblasts. Heterozygotes do not develop clavicles, but are otherwise normal. These phenotypes are indistinguishable from those of the homozygous and heterozygous null mutants, indicating that the intranuclear targeting signal is a critical determinant for function. The expressed truncated Runx2 Delta C protein enters the nucleus and retains normal DNA binding activity, but shows complete loss of intranuclear targeting. These results demonstrate that the multifunctional N-terminal region of the Runx2 protein is not sufficient for biological activity. We conclude that subnuclear localization of Runx factors in specific foci together with associated regulatory functions is essential for control of Runx-dependent genes involved in tissue differentiation during embryonic development. PMID- 11438702 TI - Energy-driven subunit rotation at the interface between subunit a and the c oligomer in the F(O) sector of Escherichia coli ATP synthase. AB - Subunit rotation within the F(1) catalytic sector of the ATP synthase has been well documented, identifying the synthase as the smallest known rotary motor. In the membrane-embedded F(O) sector, it is thought that proton transport occurs at a rotor/stator interface between the oligomeric ring of c subunits (rotor) and the single-copy a subunit (stator). Here we report evidence for an energy dependent rotation at this interface. F(O)F(1) was expressed with a pair of substituted cysteines positioned to allow an intersubunit disulfide crosslink between subunit a and a c subunit [aN214C/cM65C; Jiang, W. & Fillingame, R. H. (1998) Proc. Natl. Acad. Sci. USA 95, 6607--6612]. Membranes were treated with N,N'-dicyclohexyl-[(14)C]carbodiimide to radiolabel the D61 residue on less than 20% of the c subunits. After oxidation to form an a--c crosslink, the c subunit properly aligned to crosslink to subunit a was found to contain very little (14)C label relative to other members of the c ring. However, exposure to MgATP before oxidation significantly increased the radiolabel in the a-c crosslink, indicating that a different c subunit was now aligned with subunit a. This increase was not induced by exposure to MgADP/P(i). Furthermore, preincubation with MgADP and azide to inhibit F(1) or with high concentrations of N,N' dicyclohexylcarbodiimide to label most c subunits prevented the ATP effect. These results provide evidence for an energy-dependent rotation of the c ring relative to subunit a. PMID- 11438703 TI - Complex regulation of human inducible nitric oxide synthase gene transcription by Stat 1 and NF-kappa B. AB - The human inducible nitric oxide synthase (hiNOS) gene is expressed in several disease states and is also important in the normal immune response. Previously, we described a cytokine-responsive enhancer between -5.2 and -6.1 kb in the 5' flanking hiNOS promoter DNA, which contains multiple nuclear factor kappa beta (NF-kappa B) elements. Here, we describe the role of the IFN-Jak kinase-Stat (signal transducer and activator of transcription) 1 pathway for regulation of hiNOS gene transcription. In A549 human lung epithelial cells, a combination of cytokines tumor necrosis factor-alpha, interleukin-1 beta, and IFN-gamma (TNF alpha, IL-1 beta, and IFN-gamma) function synergistically for induction of hiNOS transcription. Pharmacological inhibitors of Jak2 kinase inhibit cytokine-induced Stat 1 DNA-binding and hiNOS gene expression. Expression of a dominant-negative mutant Stat 1 inhibits cytokine-induced hiNOS reporter expression. Site-directed mutagenesis of a cis-acting DNA element at -5.8 kb in the hiNOS promoter identifies a bifunctional NF-kappa B/Stat 1 motif. In contrast, gel shift assays indicate that only Stat 1 binds to the DNA element at -5.2 kb in the hiNOS promoter. Interestingly, Stat 1 is repressive to basal and stimulated iNOS mRNA expression in 2fTGH human fibroblasts, which are refractory to iNOS induction. Overexpression of NF-kappa B activates hiNOS promoter-reporter expression in Stat 1 mutant fibroblasts, but not in the wild type, suggesting that Stat 1 inhibits NF-kappa B function in these cells. These results indicate that both Stat 1 and NF-kappa B are important in the regulation of hiNOS transcription by cytokines in a complex and cell type-specific manner. PMID- 11438704 TI - Single-molecule spectroscopy of the beta(2) adrenergic receptor: observation of conformational substates in a membrane protein. AB - Single-molecule studies of the conformations of the intact beta(2) adrenergic receptor were performed in solution. Photon bursts from the fluorescently tagged adrenergic receptor in a micelle were recorded. A photon-burst algorithm and a Poisson time filter were implemented to characterize single molecules diffusing across the probe volume of a confocal microscope. The effects of molecular diffusion and photon number fluctuations were deconvoluted by assuming that Poisson distributions characterize the molecular occupation and photon numbers. Photon-burst size histograms were constructed, from which the source intensity distributions were extracted. Different conformations of the beta(2) adrenergic receptor cause quenching of the bound fluorophore to different extents and hence produce different photon-burst sizes. An analysis of the photon-burst histograms shows that there are at least two distinct substates for the native adrenergic membrane receptor. This behavior is in contrast to one peak observed for the dye molecule, rhodamine 6G. We test the reliability and robustness of the substate number determination by investigating the application of different binning criteria. Conformational changes associated with agonist binding result in a marked change in the distribution of photon-burst sizes. These studies provide insight into the conformational heterogeneity of G protein-coupled receptors in the presence and absence of a bound agonist. PMID- 11438705 TI - An antisense approach to phenotype-based gene cloning in Tetrahymena. AB - We report a pioneering approach using Tetrahymena thermophila that permits rapid identification of genes based on their null or hypomorphic phenotypes. This technique involves cell transformation with a library of plasmids that encode 26S ribosomal subunits containing short insertions. The insertions correspond to antisense sequences for a large number of genes. The majority of cells each acquires a single antisense sequence, which silences a single genomic locus. Because the insertion site within the ribosomal sequence is known, the silenced gene is easily amplified. We demonstrate that this approach can be used to identify genes required for dense core granule exocytosis. PMID- 11438706 TI - DNA bending by an adenine--thymine tract and its role in gene regulation. AB - To gain insight into the structural basis of DNA bending by adenine-thymine tracts (A-tracts) and their role in DNA recognition by gene-regulatory proteins, we have determined the crystal structure of the high-affinity DNA target of the cancer-associated human papillomavirus E2 protein. The three independent B-DNA molecules of the crystal structure determined at 2.2-A resolution are examples of A-tract-containing helices where the global direction and magnitude of curvature are in accord with solution data, thereby providing insights, at the base pair level, into the mechanism of DNA bending by such sequence motifs. A comparative analysis of E2-DNA conformations with respect to other structural and biochemical studies demonstrates that (i) the A-tract structure of the core region, which is not contacted by the protein, is critical for the formation of the high-affinity sequence-specific protein-DNA complex, and (ii) differential binding affinity is regulated by the intrinsic structure and deformability encoded in the base sequence of the DNA target. PMID- 11438707 TI - TAT peptide on the surface of liposomes affords their efficient intracellular delivery even at low temperature and in the presence of metabolic inhibitors. AB - To achieve an efficient intracellular drug and DNA delivery, attempts were made to target microparticulate drug carriers into cytoplasm bypassing the endocytotic pathway. TAT peptides derived from the HIV-1 TAT protein facilitate intracellular delivery of proteins and small colloidal particles. We demonstrated that relatively large drug carriers, such as 200-nm liposomes, can also be delivered into cells by TAT peptide attached to the liposome surface. Liposomes were fluorescently labeled with membranotropic rhodamine-phosphatidylethanolamine or by entrapping FITC-dextran. Incubation of fluorescent TAT liposomes with mouse Lewis lung carcinoma cells, human breast tumor BT20 cells, and rat cardiac myocyte H9C2 results in intracellular localization of certain liposomes. Steric hindrances for TAT peptide x cell interaction (attachment of TAT directly to the liposome surface without spacer or the presence of a high MW polyethylene glycol on the liposome surface) abolish liposome internalization, evidencing the importance of direct contact of TAT peptide with the cell surface. Low temperature or metabolic inhibitors, sodium azide or iodoacetamide, have little influence on the translocation of TAT liposomes into cells, confirming the energy independent character of this process. The approach may have important implications for drug delivery directly into cell cytoplasm. PMID- 11438708 TI - Old yellow enzyme: reduction of nitrate esters, glycerin trinitrate, and propylene 1,2-dinitrate. AB - The reaction of the old yellow enzyme and reduced flavins with organic nitrate esters has been studied. Reduced flavins have been found to react readily with glycerin trinitrate (GTN ) (nitroglycerin) and propylene dinitrate, with rate constants at pH 7.0, 25 degrees C of 145 M(-1)s(-1) and 5.8 M(-1)s(-1), respectively. With GTN, the secondary nitrate was removed reductively 6 times faster than the primary nitrate, with liberation of nitrite. With propylene dinitrate, on the other hand, the primary nitrate residue was 3 times more reactive than the secondary residue. In the old yellow enzyme-catalyzed NADPH dependent reduction of GTN and propylene dinitrate, ping-pong kinetics are displayed, as found for all other substrates of the enzyme. Rapid-reaction studies of mixing reduced enzyme with the nitrate esters show that a reduced enzyme--substrate complex is formed before oxidation of the reduced flavin. The rate constants for these reactions and the apparent K(d) values of the enzyme- substrate complexes have been determined and reveal that the rate-limiting step in catalysis is reduction of the enzyme by NADPH. Analysis of the products reveal that with the enzyme-catalyzed reactions, reduction of the primary nitrate in both GTN and propylene dinitrate is favored by comparison with the free-flavin reactions. This preferential positional reactivity can be rationalized by modeling of the substrates into the known crystal structure of the enzyme. In contrast to the facile reaction of free reduced flavins with GTN, reduced 5 deazaflavins have been found to react some 4--5 orders of magnitude slower. This finding implies that the chemical mechanism of the reaction is one involving radical transfers. PMID- 11438709 TI - Solution structure of the 2-amino-1- methyl-6-phenylimidazo[4,5-b]pyridine C8 deoxyguanosine adduct in duplex DNA. AB - The carcinogenic heterocyclic amine (HA) 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine (PhIP) is formed during the cooking of various meats. To enable structure/activity studies aimed at understanding how DNA damaged by a member of the HA class of compounds can ultimately lead to cancer, we have determined the first solution structure of an 11-mer duplex containing the C8-dG adduct formed by reaction with N-acetoxy-PhIP. A slow conformational exchange is observed in which the PhIP ligand either intercalates into the DNA helix by denaturing and displacing the modified base pair (main form) or is located outside the helix in a minimally perturbed B-DNA duplex (minor form). In the main base-displaced intercalation structure, the minor groove is widened, and the major groove is compressed at the lesion site because of the location of the bulky PhIP-N-methyl and phenyl ring in the minor groove; this distortion causes significant bending of the helix. The PhIP phenyl ring interacts with the phosphodiester-sugar ring backbone of the complementary strand and its fast rotation with respect to the intercalated imidazopyridine ring causes substantial distortions at this site, such as unwinding and bulging-out of the strand. The glycosidic torsion angle of the [PhIP]dG residue is syn, and the displaced guanine base is directed toward the 3' end of the modified strand. This study contributes, to our knowledge, the first structural information on the biologically relevant HA class to a growing body of knowledge about how conformational similarities and differences for a variety of types of lesions can influence protein interactions and ultimately biological outcome. PMID- 11438710 TI - Rapid induction of intraneuronal neurofibrillary tangles in apolipoprotein E deficient mice. AB - Cultured hippocampal slices prepared from apolipoprotein E-deficient mice were exposed to an inhibitor of cathepsins B and L and then processed for immunocytochemistry using antibodies against human paired helical filaments. Dense, AT8-immunopositive deposits were found in the subiculum, stratum oriens of hippocampal field CA1, and the hilus of the dentate gyrus. This distribution agrees with that described for tangles in Alzheimer's disease. The appearance of the labeled structures fell into categories that correspond to previously proposed stages in the progression of intraneuronal neurofibrillary tangles in human hippocampus. Electron microscopic analyses confirmed that microtubule disruption and twisted bundles of filaments were present in neurons in the affected areas. These results support the hypothesis that partial lysosomal dysfunction is a contributor to Alzheimer's disease and suggest a simple model for studying an important component of the disease. PMID- 11438711 TI - Identification of an autoimmune serum containing antibodies against the Barr body. AB - Transcriptional inactivation of one X chromosome in mammalian female somatic cells leads to condensation of the inactive X chromosome into the heterochromatic sex chromatin, or Barr body. Little is known about the molecular composition and structure of the Barr body or the mechanisms leading to its formation in female nuclei. Because human sera from patients with autoimmune diseases often contain antibodies against a variety of cellular components, we reasoned that some autoimmune sera may contain antibodies against proteins associated with the Barr body. Therefore, we screened autoimmune sera by immunofluorescence of human fibroblasts and identified one serum that immunostained a distinct nuclear structure with a size and nuclear localization consistent with the Barr body. The number of these structures was consistent with the number of Barr bodies expected in diploid female fibroblasts containing two to five X chromosomes. Immunostaining with the serum followed by fluorescence in situ hybridization with a probe against XIST RNA demonstrated that the major fluorescent signal from the autoantibody colocalized with XIST RNA. Further analysis of the serum showed that it stains human metaphase chromosomes and a nuclear structure consistent with the inactive X in female mouse fibroblasts. However, it does not exhibit localization to a Barr body-like structure in female mouse embryonic stem cells or in cells from female mouse E7.5 embryos. The lack of staining of the inactive X in cells from female E7.5 embryos suggests the antigen(s) may be involved in X inactivation at a stage subsequent to initiation of X inactivation. This demonstration of an autoantibody recognizing an antigen(s) associated with the Barr body presents a strategy for identifying molecular components of the Barr body and examining the molecular basis of X inactivation. PMID- 11438712 TI - Peripheral anti-A beta antibody alters CNS and plasma A beta clearance and decreases brain A beta burden in a mouse model of Alzheimer's disease. AB - Active immunization with the amyloid beta (A beta) peptide has been shown to decrease brain A beta deposition in transgenic mouse models of Alzheimer's disease and certain peripherally administered anti-A beta antibodies were shown to mimic this effect. In exploring factors that alter A beta metabolism and clearance, we found that a monoclonal antibody (m266) directed against the central domain of A beta was able to bind and completely sequester plasma A beta. Peripheral administration of m266 to PDAPP transgenic mice, in which A beta is generated specifically within the central nervous system (CNS), results in a rapid 1,000-fold increase in plasma A beta, due, in part, to a change in A beta equilibrium between the CNS and plasma. Although peripheral administration of m266 to PDAPP mice markedly reduces A beta deposition, m266 did not bind to A beta deposits in the brain. Thus, m266 appears to reduce brain A beta burden by altering CNS and plasma A beta clearance. PMID- 11438713 TI - Folliculostellate cell network: a route for long-distance communication in the anterior pituitary. AB - All higher life forms critically depend on hormones being rhythmically released by the anterior pituitary. The proper functioning of this master gland is dynamically controlled by a complex set of regulatory mechanisms that ultimately determine the fine tuning of the excitable endocrine cells, all of them heterogeneously distributed throughout the gland. Here, we provide evidence for an intrapituitary communication system by which information is transferred via the network of nonendocrine folliculostellate (FS) cells. Local electrical stimulation of FS cells in acute pituitary slices triggered cytosolic calcium waves, which propagated to other FS cells by signaling through gap junctions. Calcium wave initiation was because of the membrane excitability of FS cells, hitherto classified as silent cells. FS cell coupling could relay information between opposite regions of the gland. Because FS cells respond to central and peripheral stimuli and dialogue with endocrine cells, the form of large-scale intrapituitary communication described here may provide an efficient mechanism that orchestrates anterior pituitary functioning in response to physiological needs. PMID- 11438714 TI - Intrinsic noise in gene regulatory networks. AB - Cells are intrinsically noisy biochemical reactors: low reactant numbers can lead to significant statistical fluctuations in molecule numbers and reaction rates. Here we use an analytic model to investigate the emergent noise properties of genetic systems. We find for a single gene that noise is essentially determined at the translational level, and that the mean and variance of protein concentration can be independently controlled. The noise strength immediately following single gene induction is almost twice the final steady-state value. We find that fluctuations in the concentrations of a regulatory protein can propagate through a genetic cascade; translational noise control could explain the inefficient translation rates observed for genes encoding such regulatory proteins. For an autoregulatory protein, we demonstrate that negative feedback efficiently decreases system noise. The model can be used to predict the noise characteristics of networks of arbitrary connectivity. The general procedure is further illustrated for an autocatalytic protein and a bistable genetic switch. The analysis of intrinsic noise reveals biological roles of gene network structures and can lead to a deeper understanding of their evolutionary origin. PMID- 11438715 TI - Enhanced antitumor efficacy of a herpes simplex virus mutant isolated by genetic selection in cancer cells. AB - Replication-competent, attenuated herpes simplex virus-1 (HSV-1) derivatives that contain engineered mutations into the viral gamma 34.5 virulence gene have been used as oncolytic agents. However, as attenuated mutants often grow poorly, they may not completely destroy some tumors and surviving cancer cells simply regrow. Thus, although HSV-1 gamma 34.5 mutants can reduce the growth of human tumor xenografts in mice and have passed phase I safety studies, their efficacy is limited because they replicate poorly in many human tumor cells. Previously, we selected for a gamma 34.5 deletion mutant variant that regained the ability to replicate efficiently in tumor cells. Although this virus contains an extragenic suppressor mutation that confers enhanced growth in tumor cells, it remains attenuated. Here, we demonstrate that the suppressor virus replicates to greater levels in prostate carcinoma cells and, importantly, is a more potent inhibitor of tumor growth in an animal model of human prostate cancer than the gamma 34.5 parent virus. Thus, genetic selection in cancer cells can be used as a tool to enhance the antitumor activity of a replication-competent virus. The increased therapeutic potency of this oncolytic virus may be useful in the treatment of a wide variety of cancers. PMID- 11438716 TI - Molecular targeting of cancer: telomeres as targets. PMID- 11438717 TI - Chipping away at stem cells. PMID- 11438718 TI - Hidden robbers: the role of fungal haustoria in parasitism of plants. PMID- 11438719 TI - Cell culture forensics. PMID- 11438720 TI - Worms reveal essential functions for intermediate filaments. PMID- 11438721 TI - Dynamical hierarchy in transition states: why and how does a system climb over the mountain? AB - How a reacting system climbs through a transition state during the course of a reaction has been an intriguing subject for decades. Here we present and quantify a technique to identify and characterize local invariances about the transition state of an N-particle Hamiltonian system, using Lie canonical perturbation theory combined with microcanonical molecular dynamics simulation. We show that at least three distinct energy regimes of dynamical behavior occur in the region of the transition state, distinguished by the extent of their local dynamical invariance and regularity. Isomerization of a six-atom Lennard--Jones cluster illustrates this: up to energies high enough to make the system manifestly chaotic, approximate invariants of motion associated with a reaction coordinate in phase space imply a many-body dividing hypersurface in phase space that is free of recrossings even in a sea of chaos. The method makes it possible to visualize the stable and unstable invariant manifolds leading to and from the transition state, i.e., the reaction path in phase space, and how this regularity turns to chaos with increasing total energy of the system. This, in turn, illuminates a new type of phase space bottleneck in the region of a transition state that emerges as the total energy and mode coupling increase, which keeps a reacting system increasingly trapped in that region. PMID- 11438722 TI - Primate postcrania from the late middle Eocene of Myanmar. AB - Fossil primates have been known from the late middle to late Eocene Pondaung Formation of Myanmar since the description of Pondaungia cotteri in 1927. Three additional primate taxa, Amphipithecus mogaungensis, Bahinia pondaungensis and Myanmarpithecus yarshensis, were subsequently described. These primates are represented mostly by fragmentary dental and cranial remains. Here we describe the first primate postcrania from Myanmar, including a complete left humerus, a fragmentary right humerus, parts of left and right ulnae, and the distal half of a left calcaneum, all representing one individual. We assign this specimen to a large species of Pondaungia based on body size and the known geographic distribution and diversity of Myanmar primates. Body weight estimates of Pondaungia range from 4,000 to 9,000 g, based on humeral length, humeral midshaft diameter, and tooth area by using extant primate regressions. The humerus and ulna indicate that Pondaungia was capable of a wide variety of forelimb movements, with great mobility at the shoulder joint. Morphology of the distal calcaneus indicates that the hind feet were mobile at the transverse tarsal joint. Postcrania of Pondaungia present a mosaic of features, some shared in common with notharctine and adapine adapiforms, some shared with extant lorises and cebids, some shared with fossil anthropoids, and some unique. Overall, Pondaungia humeral and calcaneal morphology is most consistent with that of other known adapiforms. It does not support the inclusion of Pondaungia in Anthropoidea. PMID- 11438723 TI - Visualization of biochemical networks in living cells. AB - Functional annotation of novel genes can be achieved by detection of interactions of their encoded proteins with known proteins followed by assays to validate that the gene participates in a specific cellular function. We report an experimental strategy that allows for detection of protein interactions and functional assays with a single reporter system. Interactions among biochemical network component proteins are detected and probed with stimulators and inhibitors of the network. In addition, the cellular location of the interacting proteins is determined. We used this strategy to map a signal transduction network that controls initiation of translation in eukaryotes. We analyzed 35 different pairs of full-length proteins and identified 14 interactions, of which five have not been observed previously, suggesting that the organization of the pathway is more ramified and integrated than previously shown. Our results demonstrate the feasibility of using this strategy in efforts of genomewide functional annotation. PMID- 11438724 TI - Functional dynamics in the active site of the ribonuclease binase. AB - Binase, a member of a family of microbial guanyl-specific ribonucleases, catalyzes the endonucleotic cleavage of single-stranded RNA. It shares 82% amino acid identity with the well-studied protein barnase. We used NMR spectroscopy to study the millisecond dynamics of this small enzyme, using several methods including the measurement of residual dipolar couplings in solution. Our data show that the active site of binase is flanked by loops that are flexible at the 300-micros time scale. One of the catalytic residues, His-101, is located on such a flexible loop. In contrast, the other catalytic residue, Glu-72, is located on a beta-sheet, and is static. The residues Phe-55, part of the guanine base recognition site, and Tyr-102, stabilizing the base, are the most dynamic. Our findings suggest that binase possesses an active site that has a well-defined bottom, but which has sides that are flexible to facilitate substrate access/egress, and to deliver one of the catalytic residues. The motion in these loops does not change on complexation with the inhibitor d(CGAG) and compares well with the maximum k(cat) (1,500 s(-1)) of these ribonucleases. This observation indicates that the NMR-measured loop motions reflect the opening necessary for product release, which is apparently rate limiting for the overall turnover. PMID- 11438725 TI - Communication between noncontacting macromolecules. AB - We present a quantitative experimental demonstration of solvent-mediated communication between noncontacting biopolymers. We show that changes in the activity of a solvent component brought about by a conformational change in one biopolymer can result in changes in the physical properties of a second noncontacting biopolymer present in solution. Specifically, we show that the release of protons on denaturation of a donor polymer (in this case, a four stranded DNA tetraplex, iDNA) modulates the melting temperature of a noncontacting, acceptor polymer [in this case poly(A)]. In addition to such proton-mediated cross talk, we also demonstrate counterion-mediated cross talk between noncontacting biopolymers. Specifically, we show that counterion association/release on denaturation of native salmon sperm DNA (the donor polymer) can modulate the melting temperature of poly(dA) x poly(dT) (the acceptor polymer). Taken together, these two examples demonstrate how poly(A) and poly(dA) x poly(dT) can serve as molecular probes that report the pH and free salt concentrations in solution, respectively. Further, we demonstrate how such through-solvent dialogue between biopolymers that do not directly interact can be used to evaluate (in a model-free manner) association/dissociation reactions of solvent components (e.g., protons, sodium cations) with one of the two biopolymers. We propose that such through-solution dialogue is a general property of all biopolymers. As a result, such solvent-mediated cross talk should be considered when assessing reactions of multicomponent systems such as those that exist in essentially all biological processes. PMID- 11438726 TI - Mechanistic coupling of protease signaling and initiation of coagulation by tissue factor. AB - The crucial role of cell signaling in hemostasis is clearly established by the action of the downstream coagulation protease thrombin that cleaves platelet expressed G-protein-coupled protease activated receptors (PARs). Certain PARs are cleaved by the upstream coagulation proteases factor Xa (Xa) and the tissue factor (TF)--factor VIIa (VIIa) complex, but these enzymes are required at high nonphysiological concentrations and show limited recognition specificity for the scissile bond of target PARs. However, defining a physiological mechanism of PAR activation by upstream proteases is highly relevant because of the potent anti inflammatory in vivo effects of inhibitors of the TF initiation complex. Activation of substrate factor X (X) by the TF--VIIa complex is here shown to produce enhanced cell signaling in comparison to the TF--VIIa complex alone, free Xa, or Xa that is generated in situ by the intrinsic activation complex. Macromolecular assembly of X into a ternary complex of TF--VIIa--X is required for proteolytic conversion to Xa, and product Xa remains transiently associated in a TF--VIIa--Xa complex. By trapping this complex with a unique inhibitor that preserves Xa activity, we directly show that Xa in this ternary complex efficiently activates PAR-1 and -2. These experiments support the concept that proinflammatory upstream coagulation protease signaling is mechanistically coupled and thus an integrated part of the TF--VIIa-initiated coagulation pathway, rather than a late event during excessive activation of coagulation and systemic generation of proteolytic activity. PMID- 11438727 TI - Monitoring the GAP catalyzed H-Ras GTPase reaction at atomic resolution in real time. AB - The molecular reaction mechanism of the GTPase-activating protein (GAP)-catalyzed GTP hydrolysis by Ras was investigated by time resolved Fourier transform infrared (FTIR) difference spectroscopy using caged GTP (P(3)-1-(2 nitro)phenylethyl guanosine 5'-O-triphosphate) as photolabile trigger. This approach provides the complete GTPase reaction pathway with time resolution of milliseconds at the atomic level. Up to now, one structural model of the GAP x Ras x GDP x AlF(x) transition state analog is known, which represents a "snap shot" along the reaction-pathway. As now revealed, binding of GAP to Ras x GTP shifts negative charge from the gamma to beta phosphate. Such a shift was already identified by FTIR in GTP because of Ras binding and is now shown to be enhanced by GAP binding. Because the charge distribution of the GAP x Ras x GTP complex thus resembles a more dissociative-like transition state and is more like that in GDP, the activation free energy is reduced. An intermediate is observed on the reaction pathway that appears when the bond between beta and gamma phosphate is cleaved. In the intermediate, the released P(i) is strongly bound to the protein and surprisingly shows bands typical of those seen for phosphorylated enzyme intermediates. All these results provide a mechanistic picture that is different from the intrinsic GTPase reaction of Ras. FTIR analysis reveals the release of P(i) from the protein complex as the rate-limiting step for the GAP-catalyzed reaction. The approach presented allows the study not only of single proteins but of protein-protein interactions without intrinsic chromophores, in the non crystalline state, in real time at the atomic level. PMID- 11438728 TI - Cytochrome c' folding triggered by electron transfer: fast and slow formation of four-helix bundles. AB - Reduced (Fe(II)) Rhodopseudomonas palustris cytochrome c' (Cyt c') is more stable toward unfolding ([GuHCl](1/2) = 2.9(1) M) than the oxidized (Fe(III)) protein ([GuHCl](1/2) = 1.9(1) M). The difference in folding free energies (Delta Delta G(f) degrees = 70 meV) is less than half of the difference in reduction potentials of the folded protein (100 mV vs. NHE) and a free heme in aqueous solution ( approximately -150 mV). The spectroscopic features of unfolded Fe(II) Cyt c' indicate a low-spin heme that is axially coordinated to methionine sulfur (Met-15 or Met-25). Time-resolved absorption measurements after CO photodissociation from unfolded Fe(II)(CO)-Cyt c' confirm that methionine can bind to the ferroheme on the microsecond time scale [k(obs) = 5(2) x 10(4) s( 1)]. Protein folding was initiated by photoreduction (two-photon laser excitation of NADH) of unfolded Fe(III)-Cyt c' ([GuHCl] = 2.02--2.54 M). Folding kinetics monitored by heme absorption span a wide time range and are highly heterogeneous; there are fast-folding ( approximately 10(3) s(-1)), intermediate-folding (10(2) 10(1) s(-1)), and slow-folding (10(-1) s(-1)) populations, with the last two likely containing methionine-ligated (Met-15 or Met-25) ferrohemes. Kinetics after photoreduction of unfolded Fe(III)-Cyt c' in the presence of CO are attributable to CO binding [1.4(6) x 10(3) s(-1)] and Fe(II)(CO)-Cyt c' folding [2.8(9) s(-1)] processes; stopped-flow triggered folding of Fe(III)-Cyt c' (which does not contain a protein-derived sixth ligand) is adequately described by a single kinetics phase with an estimated folding time constant of approximately 4 ms [Delta G(f) degrees = -33(3) kJ mol(-1)] at zero denaturant. PMID- 11438729 TI - Mechanical behavior in living cells consistent with the tensegrity model. AB - Alternative models of cell mechanics depict the living cell as a simple mechanical continuum, porous filament gel, tensed cortical membrane, or tensegrity network that maintains a stabilizing prestress through incorporation of discrete structural elements that bear compression. Real-time microscopic analysis of cells containing GFP-labeled microtubules and associated mitochondria revealed that living cells behave like discrete structures composed of an interconnected network of actin microfilaments and microtubules when mechanical stresses are applied to cell surface integrin receptors. Quantitation of cell tractional forces and cellular prestress by using traction force microscopy confirmed that microtubules bear compression and are responsible for a significant portion of the cytoskeletal prestress that determines cell shape stability under conditions in which myosin light chain phosphorylation and intracellular calcium remained unchanged. Quantitative measurements of both static and dynamic mechanical behaviors in cells also were consistent with specific a priori predictions of the tensegrity model. These findings suggest that tensegrity represents a unified model of cell mechanics that may help to explain how mechanical behaviors emerge through collective interactions among different cytoskeletal filaments and extracellular adhesions in living cells. PMID- 11438730 TI - Configurational diffusion down a folding funnel describes the dynamics of DNA hairpins. AB - Elucidating the mechanism of folding of polynucleotides depends on accurate estimates of free energy surfaces and a quantitative description of the kinetics of structure formation. Here, the kinetics of hairpin formation in single stranded DNA are measured after a laser temperature jump. The kinetics are modeled as configurational diffusion on a free energy surface obtained from a statistical mechanical description of equilibrium melting profiles. The effective diffusion coefficient is found to be strongly temperature-dependent in the nucleation step as a result of formation of misfolded loops that do not lead to subsequent zipping. This simple system exhibits many of the features predicted from theoretical studies of protein folding, including a funnel-like energy surface with many folding pathways, trapping in misfolded conformations, and non Arrhenius folding rates. PMID- 11438731 TI - Polarity and permeation profiles in lipid membranes. AB - The isotropic (14)N-hyperfine coupling constant, a(o)(N), of nitroxide spin labels is dependent on the local environmental polarity. The dependence of a(o)(N) in fluid phospholipid bilayer membranes on the C-atom position, n, of the nitroxide in the sn-2 chain of a spin-labeled diacyl glycerophospholipid therefore determines the transmembrane polarity profile. The polarity variation in phospholipid membranes, with and without equimolar cholesterol, is characterized by a sigmoidal, trough-like profile of the form (1 + exp [(n - n(o))/lambda])(-1), where n = n(o) is the point of maximum gradient, or polarity midpoint, beyond which the free energy of permeation decreases linearly with n, on a characteristic length-scale, lambda. Integration over this profile yields a corresponding expression for the permeability barrier to polar solutes. For fluid membranes without cholesterol, n(o) approximately 8 and lambda approximately 0.5- 1 CH(2) units, and the permeability barrier introduces an additional diffusive resistance that is equivalent to increasing the effective membrane thickness by 35--80%, depending on the lipid. For membranes containing equimolar cholesterol, n(o) approximately 9--10, and the total change in polarity is greater than for membranes without cholesterol, increasing the permeability barrier by a factor of 2, whereas the decay length remains similar. The permeation of oxygen into fluid lipid membranes (determined by spin-label relaxation enhancements) displays a profile similar to that of the transmembrane polarity but of opposite sense. For fluid membranes without cholesterol n(o) approximately 8 and lambda approximately 1 CH(2) units, also for oxygen. The permeation profile for polar paramagnetic ion complexes is closer to a single exponential decay, i.e., n(o) lies outside the acyl-chain region of the membrane. These results are relevant not only to the permeation of water and polar solutes into membranes and their permeabilities, but also to depth determinations of site-specifically spin-labeled protein residues by using paramagnetic relaxation agents. PMID- 11438732 TI - Repression of TFII-I-dependent transcription by nuclear exclusion. AB - TFII-I is an unusual transcription factor possessing both basal and signal induced transcriptional functions. Here we report the characterization of a TFII I-related factor (MusTRD1/BEN) that regulates transcriptional functions of TFII-I by controlling its nuclear residency. MusTRD1/BEN has five or six direct repeats, each containing helix--loop--helix motifs, and, thus, belongs to the TFII-I family of transcription factors. TFII-I and MusTRD1/BEN, when expressed individually, show predominant nuclear localization. However, when the two proteins are coexpressed ectopically, MusTRD1/BEN locates almost exclusively to the nucleus, whereas TFII-I is largely excluded from the nucleus, resulting in a loss of TFII-I-dependent transcriptional activation of the c-fos promoter. Mutation of a consensus nuclear localization signal in MusTRD1/BEN results in a reversal of nuclear residency of the two proteins and a concomitant gain of c-fos promoter activity. These data suggest a means of transcriptional repression by competition at the level of nuclear occupancy. PMID- 11438733 TI - Clues to epidermal cancer proneness revealed by reconstruction of DNA repair deficient xeroderma pigmentosum skin in vitro. AB - Sun exposure has been clearly implicated in premature skin aging and neoplastic development. These features are exacerbated in patients with xeroderma pigmentosum (XP), a hereditary disease, the biochemical hallmark of which is a severe deficiency in the nucleotide excision repair of UV-induced DNA lesions. To develop an organotypic model of DNA repair deficiency, we have cultured several strains of primary XP keratinocytes and XP fibroblasts from skin biopsies of XP patients. XP skin comprising both a full-thickness epidermis and a dermal equivalent was successfully reconstructed in vitro. Satisfactory features of stratification were obtained, but the expression of epidermal differentiation products, such as keratin K10 and loricrin, was delayed and reduced. In addition, the proliferation of XP keratinocytes was more rapid than that of normal keratinocytes. Moreover, increased deposition of cell attachment proteins, alpha 6 and beta-1 integrins, was observed in the basement membrane zone, and beta-1 integrin subunit, the expression of which is normally confined to basal keratinocytes, extended into several suprabasal cell layers. Most strikingly, the in vitro reconstructed XP skin displayed numerous proliferative epidermal invasions within dermal equivalents. Epidermal invasion and higher proliferation rate are reminiscent of early steps of neoplasia. Compared with normal skin, the DNA repair deficiency of in vitro reconstructed XP skin was documented by long lasting persistence of UVB-induced DNA damage in all epidermal layers, including the basal layer from which carcinoma develops. The availability of in vitro reconstructed XP skin provides opportunities for research in the fields of photoaging, photocarcinogenesis, and tissue therapy. PMID- 11438734 TI - Guanylyl cyclase C agonists regulate progression through the cell cycle of human colon carcinoma cells. AB - The effects of Escherichia coli heat-stable enterotoxin (ST) and uroguanylin were examined on the proliferation of T84 and Caco2 human colon carcinoma cells that express guanylyl cyclase C (GC-C) and SW480 human colon carcinoma cells that do not express this receptor. ST or uroguanylin inhibited proliferation of T84 and Caco2 cells, but not SW480 cells, in a concentration-dependent fashion, assessed by quantifying cell number, cell protein, and [(3)H]thymidine incorporation into DNA. These agonists did not inhibit proliferation by induction of apoptosis, assessed by TUNEL (terminal deoxynucleotidyl transferase-mediated dNTP-biotin nick end labeling of DNA fragments) assay and DNA laddering, or necrosis, assessed by trypan blue exclusion and lactate dehydrogenase release. Rather, ST prolonged the cell cycle, assessed by flow cytometry and [(3)H]thymidine incorporation into DNA. The cytostatic effects of GC-C agonists were associated with accumulation of intracellular cGMP, mimicked by the cell-permeant analog 8 Br-cGMP, and reproduced and potentiated by the cGMP-specific phosphodiesterase inhibitor zaprinast but not the inactive ST analog TJU 1-103. Thus, GC-C agonists regulate the proliferation of intestinal cells through cGMP-dependent mechanisms by delaying progression of the cell cycle. These data suggest that endogenous agonists of GC-C, such as uroguanylin, may play a role in regulating the balance between epithelial proliferation and differentiation in normal intestinal physiology. Therefore, GC-C ligands may be novel therapeutic agents for the treatment of patients with colorectal cancer. PMID- 11438735 TI - Role of the iroquois3 homeobox gene in organizer formation. AB - In zebrafish, the organizer is thought to consist of two regions, the yolk syncytial layer (YSL) and the shield. The dorsal YSL appears to send signals that affect formation of the shield in the overlying mesendoderm. We show here that a domain of dorsal deep cells located between the YSL and the shield is marked by expression of the iro3 gene. As gastrulation proceeds, the iro3 positive domain involutes and migrates to the animal pole. Iro3 expression is regulated by Nodal and bone morphogenic protein antagonists. Overexpression of iro3 induced ectopic expression of shield-specific genes. This effect was mimicked by an Iro3 Engrailed transcriptional repressor domain fusion, whereas an Iro3-VP16 activator domain fusion behaved as a dominant negative or antimorphic form. These results suggest that Iro3 acts as a transcriptional repressor and further implicate the iro3 gene in regulating organizer formation. We propose that the iro3-expressing dorsal deep cells represent a distinct organizer domain that receives signals from the YSL and in turn sends signals to the forming shield, thereby influencing its expansion and differentiation. PMID- 11438736 TI - The cranium of Parapithecus grangeri, an Egyptian Oligocene anthropoidean primate. AB - A nearly complete skull of Parapithecus grangeri from the early Oligocene of Egypt is described. The specimen is relatively undistorted and is undoubtedly the most complete higher primate skull yet found in the African Oligocene, which also makes it the most complete Oligocene primate cranium worldwide. Belonging in superfamily Parapithecoidea, a group regarded by some as the sister group to all other Anthropoidea, this skull reveals important information about the radiation of stem anthropoideans. This cranium is about 15% larger than size estimates based on a fragmentary cranium of its contemporary and close relative Apidium phiomense. It is about the same size as that of the gray gentle lemur, Hapalemur griseus, or of platyrrhines such as the owl monkey, Aotus trivirgatus, or the titi monkey, Callicebus torquatus. Comparatively small orbits and size differences in jaws and teeth show it was both diurnal and dimorphic. This is the only specimen of the species that shows (from sockets) that there were four small upper incisors. Several mandibular specimens of the species establish that there were no permanent lower incisors and that the symphysis was fused. Like other early anthropoideans this species possessed a lower encephalization quotient and less-developed orbital frontality than later anthropoideans. There is full postorbital closure and fusion of the metopic suture, and the ectotympanic forms a rim to the auditory aperture. A probable frontal/alisphenoid contact is a potentially derived resemblance to Catarrhini. A proposed separate genus for the species P. grangeri is not sustained. PMID- 11438737 TI - A mouse model for adenovirus gene delivery. AB - The cellular attachment receptor for adenovirus (Ad), Coxsackie adenovirus receptor (CAR), required for delivery of Ad into primary cells, is not present on all cell types, thus restricting Ad-gene delivery systems. To circumvent this constrain, a transgenic mouse has been generated that expresses a truncated human CAR in all tissues analyzed. These mice allowed efficient in vitro infections at low multiplicities into lymphoid, myeloid, and endothelial cells. Furthermore, in vivo administration of Ad-vectors results in infection of macrophages, lymphocytes, and endothelial cells. In addition, tail vein injection resulted in targeting of virus into previously inaccessible areas, such as the lung and the capillaries of the brain. The CAR transgenic mice will be useful for rapid functional genomic analysis in vivo, for testing the efficacy of gene therapy procedures or as a source of easily transducible cells. PMID- 11438738 TI - From hematopoiesis to neuropoiesis: evidence of overlapping genetic programs. AB - It is reasonable to propose that gene expression profiles of purified stem cells could give clues for the molecular mechanisms of stem cell behavior. We took advantage of cDNA subtraction to identify a set of genes selectively expressed in mouse adult hematopoietic stem cells (HSC) as opposed to bone marrow (BM). Analysis of HSC-enriched genes revealed several key regulatory gene candidates, including two novel seven transmembrane (7TM) receptors. Furthermore, by using cDNA microarray techniques we found a large set of HSC-enriched genes that are expressed in mouse neurospheres (a population greatly enriched for neural progenitor cells), but not present in terminally differentiated neural cells. In situ hybridization demonstrated that many of them, including one HSC-enriched 7TM receptor, were selectively expressed in the germinal zones of fetal and adult brain, the regions harboring mouse neural stem cells. We propose that at least some of the transcripts that are selectively and commonly expressed in two or more types of stem cells define a functionally conserved group of genes evolved to participate in basic stem cell functions, including stem cell self-renewal. PMID- 11438739 TI - Genes linked by fusion events are generally of the same functional category: a systematic analysis of 30 microbial genomes. AB - Recent work in computational genomics has shown that a functional association between two genes can be derived from the existence of a fusion of the two as one continuous sequence in another genome. For each of 30 completely sequenced microbial genomes, we established all such fusion links among its genes and determined the distribution of links within and among 15 broad functional categories. We found that 72% of all fusion links related genes of the same functional category. A comparison of the distribution of links to simulations on the basis of a random model further confirmed the significance of intracategory fusion links. Where a gene of annotated function is linked to an unclassified gene, the fusion link suggests that the two genes belong to the same functional category. The predictions based on fusion links are shown here for Methanobacterium thermoautotrophicum, and another 661 predictions are available at http://fusion.bu.edu. PMID- 11438740 TI - Localization of atherosclerosis susceptibility loci to chromosomes 4 and 6 using the Ldlr knockout mouse model. AB - Atherosclerosis is a complex disease resulting from the interaction of multiple genes. We have used the Ldlr knockout mouse model in an interspecific genetic cross to map atherosclerosis susceptibility loci. A total of 174 (MOLF/Ei x B6.129S7-Ldlr(tm1Her)) x C57BL/6J-Ldlr(tm1Her) backcross mice, homozygous for the Ldlr null allele, were fed a Western-type diet for 3 months and then killed for quantification of aortic lesions. A genome scan was carried out by using DNA pools and microsatellite markers spaced at approximately 18-centimorgan intervals. Quantitative trait locus analysis of individual backcross mice confirmed linkages to chromosomes 4 (Athsq1, logarithm of odds = 6.2) and 6 (Athsq2, logarithm of odds = 6.7). Athsq1 affected lesions in females only whereas Athsq2 affected both sexes. Among females, the loci accounted for approximately 50% of the total variance of lesion area. The susceptible allele at Athsq1 was derived from the MOLF/Ei genome whereas the susceptible allele at Athsq2 was derived from C57BL/6J. Inheritance of susceptible alleles at both loci conferred a 2-fold difference in lesion area, suggesting an additive effect of Athsq1 and Athsq2. No associations were observed between the quantitative trait loci and levels of plasma total cholesterol, high density lipoprotein cholesterol, non-high density lipoprotein cholesterol, insulin, or body weight. We provide strong evidence for complex inheritance of atherosclerosis in mice with elevated plasma low density lipoprotein cholesterol and show a major influence of nonlipoprotein-related factors on disease susceptibility. Athsq1 and Athsq2 represent candidate susceptibility loci for human atherosclerosis, most likely residing on chromosomes 1p36--32 and 12p13--12, respectively. PMID- 11438741 TI - Quantitative analysis of chromosomal CGH in human breast tumors associates copy number abnormalities with p53 status and patient survival. AB - We present a general method for rigorously identifying correlations between variations in large-scale molecular profiles and outcomes and apply it to chromosomal comparative genomic hybridization data from a set of 52 breast tumors. We identify two loci where copy number abnormalities are correlated with poor survival outcome (gain at 8q24 and loss at 9q13). We also identify a relationship between abnormalities at two loci and the mutational status of p53. Gain at 8q24 and loss at 5q15-5q21 are linked with mutant p53. The 9q and 5q losses suggest the possibility of gene products involved in breast cancer progression. The analytical techniques are general and also are applicable to the analysis of array-based expression data. PMID- 11438742 TI - Chemokine receptor 2 serves an early and essential role in resistance to Mycobacterium tuberculosis. AB - Although the protective cellular immune response to Mycobacterium tuberculosis requires recruitment of macrophages and T lymphocytes to the site of infection, the signals that regulate this trafficking have not been defined. We investigated the role of C-C chemokine receptor 2 (CCR2)-dependent cell recruitment in the protective response to M. tuberculosis. CCR2(-/-) mice died early after infection and had 100-fold more bacteria in their lungs than did CCR2(+/+) mice. CCR2(-/-) mice exhibited an early defect in macrophage recruitment to the lung and a later defect in recruitment of dendritic cells and T cells to the lung. CCR2(-/-) mice also had fewer macrophages and dendritic cells recruited to the mediastinal lymph node (MLN) after infection. T cell migration through the MLN was similar in CCR2( /-) and CCR2(+/+) mice. However, T cell priming was delayed in the MLNs of the CCR2(-/-) mice, and fewer CD4(+) and CD8(+) T cells primed to produce IFN-gamma accumulated in the lungs of the CCR2(-/-) mice. These data demonstrate that cellular responses mediated by activation of CCR2 are essential in the initial immune response and control of infection with M. tuberculosis. PMID- 11438743 TI - Cysteinyl leukotriene receptor 1 is also a pyrimidinergic receptor and is expressed by human mast cells. AB - The cysteinyl leukotrienes (cys-LTs) LTC(4), LTD(4), and LTE(4) are a class of peptide-conjugated lipids formed from arachidonic acid and released during activation of mast cells (MCs). We now report that human cord-blood-derived MCs (hMCs) express the CysLT1 receptor, which responds not only to inflammation derived cys-LTs, but also to a pyrimidinergic ligand, UDP. hMCs express both CysLT1 protein and transcript, and respond to LTC(4), LTD(4), and UDP with concentration-dependent calcium fluxes, each of which is blocked by a competitive CysLT1 receptor antagonist, MK571. Stably transfected Chinese hamster ovary cells expressing the CysLT1 receptor also exhibit MK571-sensitive calcium flux to all three agonists. Both hMCs and CysLT1 transfectants stimulated with UDP are desensitized to LTC(4), but only partially to LTD(4). Priming of hMCs with IL-4 for 5 days enhances their sensitivity to each agonist, but preferentially lowers their threshold for activation by LTC(4) and UDP (approximately 3 log(10)-fold shifts in dose-response for each agonist) over LTD(4) (1.3 log(10)-fold shift), without altering CysLT1 receptor mRNA or surface protein expression, implying the likely induction of a second receptor with CysLT1-like dual ligand specificity. hMCs thus express the CysLT1 receptor, and possibly a closely related IL-4 inducible receptor, which mediate dual activation responses to cys-LTs and UDP, providing an apparent intersection linking the inflammatory and neurogenic elements of bronchial asthma. PMID- 11438744 TI - A low threshold level of expression of mutant-template telomerase RNA inhibits human tumor cell proliferation. AB - The ribonucleoprotein telomerase synthesizes telomeric DNA by copying an intrinsic RNA template. In most cancer cells, telomerase is highly activated. Here we report a telomerase-based antitumor strategy: expression of mutant template telomerase RNAs in human cancer cells. We expressed mutant-template human telomerase RNAs in prostate (LNCaP) and breast (MCF-7) cancer cell lines. Even a low threshold level of expression of telomerase RNA gene constructs containing various mutant templates, but not the control wild-type template, decreased cellular viability and increased apoptosis. This occurred despite the retention of normal levels of the endogenous wild-type telomerase RNA and endogenous wild-type telomerase activity and unaltered stable telomere lengths. In vivo tumor xenografts of a breast cancer cell line expressing a mutant template telomerase RNA also had decreased growth rates. Therefore, mutant template telomerase RNAs exert a strongly dominant-negative effect on cell proliferation and tumor growth. These results support the potential use of mutant template telomerase RNA expression as an antineoplastic strategy. PMID- 11438745 TI - C/EBP epsilon mediates myeloid differentiation and is regulated by the CCAAT displacement protein (CDP/cut). AB - Neutrophils from CCAAT enhancer binding protein epsilon (C/EBP epsilon) knockout mice have morphological and biochemical features similar to those observed in patients with an extremely rare congenital disorder called neutrophil-specific secondary granule deficiency (SGD). SGD is characterized by frequent bacterial infections attributed, in part, to the lack of neutrophil secondary granule proteins (SGP). A mutation that results in loss of functional C/EBP epsilon activity has recently been described in an SGD patient, and has been postulated to be the cause of the disease in this patient. We have previously demonstrated that overexpression of CCAAT displacement protein (CDP/cut), a highly conserved transcriptional repressor of developmentally regulated genes, suppresses expression of SGP genes in 32Dcl3 cells. This phenotype resembles that observed in both C/EBP epsilon(-/-) mice and in SGD patients. Based on these observations we investigated potential interactions between C/EBP epsilon and CDP/cut during neutrophil maturation. In this study, we demonstrate that inducible expression of C/EBP epsilon in 32Dcl3/tet cells results in granulocytic differentiation. Furthermore, Northern blot analysis of G-CSF-induced CDP/cut overexpressing 32Dcl3 cells revealed absence of C/EBP epsilon mRNA. We therefore hypothesize that C/EBP epsilon positively regulates SGP gene expression, and that C/EBP epsilon is itself negatively regulated by CDP/cut during neutrophil maturation. We further demonstrate that the C/EBP epsilon promoter is regulated by CDP/cut during myeloid differentiation. PMID- 11438746 TI - Toward a new generation of vaccines: the anti-cytokine therapeutic vaccines. AB - Pathological conditions, such as cancers, viral infections, and autoimmune diseases, are associated with abnormal cytokine production, and the morbidity associated with many medical disorders is often directly a result of cytokine production. Because of the absence of negative feedback control occurring in some pathophysiologic situations, a given cytokine may flood and accumulate in the extracellular compartment of tissues or tumors thereby impairing the cytokine network homeostasis and contributing to local pathogenesis. To evaluate whether the rise of anti-cytokine Abs by vaccination is an effective way to treat these pathological conditions without being harmful to the organism, we have analyzed each step of the cytokine process (involving cytokine production, target response, and feedback regulation) and have considered them in the local context of effector--target cell microenvironment and in the overall context of the macroenvironment of the immune system of the organism. In pathologic tissues, Abs of high affinity, as raised by anti-cytokine vaccination, should neutralize the pool of cytokines ectopically accumulated in the extracellular compartment, thus counteracting their pathogenic effects. In contrast, the same Abs should not interfere with cytokine processes occurring in normal tissues, because under physiologic conditions cytokine production by effector cells (induced by activation but controlled by negative feedback regulation) does not accumulate in the extracellular compartment. These concepts are consistent with results showing that following animal and human anti-cytokine vaccination, induction of high affinity Abs has proven to be safe and effective and encourages this approach as a pioneering avenue of therapy. PMID- 11438747 TI - Modular organization of intrinsic connections associated with spectral tuning in cat auditory cortex. AB - Many response properties in primary auditory cortex (AI) are segregated spatially and organized topographically as those in primary visual cortex. Intensive study has not revealed an intrinsic, anatomical organizing principle related to an AI functional topography. We used retrograde anatomic tracing and topographic physiologic mapping of acoustic response properties to reveal long-range (> or = 1.5 mm) convergent intrinsic horizontal connections between AI subregions with similar bandwidth and characteristic frequency selectivity. This suggests a modular organization for processing spectral bandwidth in AI. PMID- 11438748 TI - Brain activity evidence for recognition without recollection after early hippocampal damage. AB - Amnesic patients with early and seemingly isolated hippocampal injury show relatively normal recognition memory scores. The cognitive profile of these patients raises the possibility that this recognition performance is maintained mainly by stimulus familiarity in the absence of recollection of contextual information. Here we report electrophysiological data on the status of recognition memory in one of the patients, Jon. Jon's recognition of studied words lacks the event-related potential (ERP) index of recollection, viz., an increase in the late positive component (500--700 ms), under conditions that elicit it reliably in normal subjects. On the other hand, a decrease of the ERP amplitude between 300 and 500 ms, also reliably found in normal subjects, is well preserved. This so-called N400 effect has been linked to stimulus familiarity in previous ERP studies of recognition memory. In Jon, this link is supported by the finding that his recognized and unrecognized studied words evoked topographically distinct ERP effects in the N400 time window. These data suggest that recollection is more dependent on the hippocampal formation than is familiarity, consistent with the view that the hippocampal formation plays a special role in episodic memory, for which recollection is so critical. PMID- 11438749 TI - BDNF but not NT-4 is required for normal flexion reflex plasticity and function. AB - Neurotrophins can directly modulate the function of diverse types of central nervous system synapses. Brain-derived neurotrophic factor (BDNF) might be released by nociceptors onto spinal neurons and mediate central sensitization associated with chronic pain. We have studied the role of BDNF and neurotrophin-4 (NT-4), both ligands of the trkB tyrosine kinase receptor, in synaptic transmission and reflex plasticity in the mouse spinal cord. We used an in vitro spinal cord preparation to measure monosynaptic and polysynaptic reflexes evoked by primary afferents in BDNF- and NT-4-deficient mice. In situ hybridization studies show that both these neurotrophins are synthesized by sensory neurons, and NT-4, but not BDNF, also is expressed by spinal neurons. BDNF null mutants display selective deficits in the ventral root potential (VRP) evoked by stimulating nociceptive primary afferents whereas the non-nociceptive portion of the VRP remained unaltered. In addition, activity-dependent plasticity of the VRP evoked by repetitive (1 Hz) stimulation of nociceptive primary afferents (termed wind-up) was substantially reduced in BDNF-deficient mice. This plasticity also was reduced in a reversible manner by the protein kinase inhibitor K252a. Although the trkB ligand NT-4 is normally present, reflex properties in NT-4 null mutant mice were normal. Pharmacological studies also indicated that spinal N methyl-d-aspartate receptor function was unaltered in BDNF-deficient mice. Using immunocytochemistry for markers of nociceptive neurons we found no evidence that their number or connectivity was substantially altered in BDNF-deficient mice. Our data therefore are consistent with a direct role for presynaptic BDNF release from sensory neurons in the modulation of pain-related neurotransmission. PMID- 11438750 TI - The synthesis, discovery, and development of a highly promising class of microtubule stabilization agents: curative effects of desoxyepothilones B and F against human tumor xenografts in nude mice. AB - We have evaluated two synthetic epothilone analogues lacking the 12,13-epoxide functionality, 12,13-desoxyepothilone B (dEpoB), and 12,13-desoxyepothilone F (dEpoF). The concentrations required for 50% growth inhibition (IC(50)) for a variety of anticancer agents were measured in CCRF-CEM/VBL1000 cells (2,048-fold resistance to vinblastine). By using dEpoB, dEpoF, aza-EpoB, and paclitaxel, the IC(50) values were 0.029, 0.092, 2.99, and 5.17 microM, respectively. These values represent 4-, 33.5-, 1,423- and 3,133-fold resistance, respectively, when compared with the corresponding IC(50) in the parent [nonmultiple drug-resistant (MDR)] CCRF-CEM cells. We then produced MDR human lung carcinoma A549 cells by continuous exposure of the tumor cells to sublethal concentrations of dEpoB (1.8 yr), vinblastine (1.2 yr), and paclitaxel (1.8 yr). This continued exposure led to the development of 2.1-, 4,848-, and 2,553-fold resistance to each drug, respectively. The therapeutic effect of dEpoB and paclitaxel was also compared in vivo in a mouse model by using various tumor xenografts. dEpoB is much more effective in reducing tumor sizes in all MDR tumors tested. Analysis of dEpoF, an analog possessing greater aqueous solubility than dEpoB, showed curative effects similar to dEpoB against K562, CCRF-CEM, and MX-1 xenografts. These results indicate that dEpoB and dEpoF are efficacious antitumor agents with both a broad chemotherapeutic spectrum and wide safety margins. PMID- 11438751 TI - Parallel detection of violations of color constancy. AB - The perceived colors of reflecting surfaces generally remain stable despite changes in the spectrum of the illuminating light. This color constancy can be measured operationally by asking observers to distinguish illuminant changes on a scene from changes in the reflecting properties of the surfaces comprising it. It is shown here that during fast illuminant changes, simultaneous changes in spectral reflectance of one or more surfaces in an array of other surfaces can be readily detected almost independent of the numbers of surfaces, suggesting a preattentive, spatially parallel process. This process, which is perfect over a spatial window delimited by the anatomical fovea, may form an early input to a multistage analysis of surface color, providing the visual system with information about a rapidly changing world in advance of the generation of a more elaborate and stable perceptual representation. PMID- 11438752 TI - Enhancement of enzyme activity through three-phase partitioning: crystal structure of a modified serine proteinase at 1.5 A resolution. AB - Three-phase partitioning is fast developing as a novel bioseparation strategy with a wide range of applications including enzyme stability and enhancement of its catalytic activity. Despite all this, the enzyme behaviour in this process still remains unknown. A serine proteinase, proteinase K, was subjected to three phase partitioning (TPP). A 3 ml volume of proteinase K solution (3 mg/ml in 0.05 M acetate buffer, pH 6.0) was brought to 30% (w/v) ammonium sulphate saturation by addition of saturated ammonium sulphate. tert-Butanol (6 ml) was added to this solution and the mixture was incubated at 25 degrees C for 1 h. The precipitated protein in the mid-layer was dissolved in 3 ml of 0.05 M acetate buffer, pH 6.0. The specific activity of the processed enzyme was estimated and was found to be 210% of the original enzyme activity. In order to understand the basis of this remarkable enhancement of the enzyme activity, the structure of the TPP-treated enzyme was determined by X-ray diffraction at 1.5 A resolution. The overall structure of the TPP-treated enzyme is similar to the original structure in an aqueous environment. The hydrogen bonding system of the catalytic triad is intact. However, the water structure in the substrate binding site has undergone a rearrangement as some of the water molecules are either displaced or completely absent. Two acetate ions were identified in the structure. One is located in the active site and seems to mimic the role of water in the enzyme activity and stability. The other is located at the surface of the molecule and is involved in stabilizing the local structure of the enzyme. The most striking observation in respect of the present structure pertains to a relatively higher overall temperature factor (B = 19.7 A(2)) than the value of 9.3 A(2) in the original enzyme. As a result of a higher B-factor, a number of residues, particularly their side chains, were found to adopt more than one conformation. It appears that the protein exists in an excited state which might be helping the enzyme to function more rapidly than the original enzyme in aqueous media. Summarily, the basis of increased enzymatic activity could be attributed to (i) the presence of an acetate ion at the active site and (ii) its excited state as reflected by an overall higher B-factor. PMID- 11438753 TI - Organization of regions with amphiphilic alpha-helical potential within the three dimensional structure of beta-sheet proteins. AB - The observation that strong amphiphilic alpha-helical potential exists in all proteins, including beta-sheet proteins, has given rise to the idea that alpha helical intermediates may be critical to the folding paths of all proteins. Here we report that regions with amphiphilic alpha-helical potential in beta-sheet proteins are regularly spaced within the native structure of the proteins at an average interval of about 13 A. This regular spacing did not occur when the location of amphiphilic regions was randomly assigned (p = 0.0056), suggesting some degree of organization with respect to the native fold. However, in the native structure of various non-homologous proteins that contain the same fold, the location of the regions with amphiphilic alpha-helical potential was not conserved. Further, there was no apparent association of amphiphilic alpha helical potential with any particular type of secondary structure, confirming that this potential is not involved in maintenance of native structure and suggesting that it may be associated with a highly adaptable process. PMID- 11438754 TI - Side-chain conformations in 4-alpha-helical bundles. AB - The distribution of the chi(1), chi(2) dihedral angles in a dataset consisting of 12 unrelated 4-alpha-helical bundle proteins was determined and qualitatively compared with that observed in globular proteins. The analysis suggests that the 4-alpha-helical bundle motif could occasionally impose steric constraints on side chains: (i) the side-chain conformations are limited to only a subset of the conformations observed in globular proteins and for some amino acids they are sterically more constrained than those in helical regions of globular proteins; (ii) aspartic acid and asparagine occasionally adopt rotamers that have not been previously reported for globular or helical proteins; (iii) some rotamers of tyrosine and isoleucine are predominantly or exclusively associated with hydrophobic core positions (a, d); (iv) mutations in the hydrophobic core occur preferentially between residue types which among other physicochemical properties also share a predominant rotamer. PMID- 11438755 TI - Solvent entropy-driven searching for protein modeling examined and tested in simplified models. AB - Solvent entropy is a force to consider in protein folding and protein design but is difficult to model. It is investigated here in the context of the hp model: Two types of residues, hydrophobic and hydrophilic, are modeled on a lattice. Nine chains and two- and three-dimensional simulations are compared. We show that considering solvent entropy alone, efficient folding of lattice chains (identification of the native fold) can be achieved by an entropy-driven simulation on its own. Moreover, in a detailed comparison over a wide range of parameters, entropy-guided searching outperforms an energy-driven search in the model. The combination of energy- and entropy-driven search yields the most efficient searching. It is compared in detail with the above results, indicating also how this solvent shell model may advantageously be implemented in more complex protein modeling simulations. PMID- 11438756 TI - A three-dimensional model of endothelin-converting enzyme (ECE) based on the X ray structure of neutral endopeptidase 24.11 (NEP). AB - Endothelin-converting enzyme 1 (ECE-1, EC 3.4.24.71) is a zinc-dependent type II mammalian membrane protein comprising the active site in the ectodomain. It exists in multiple splice variants that all catalyze the last and rate-limiting step in the activation of preproendothelin to the highly potent vasoconstrictor endothelin. There is high interest in finding small and potent inhibitors for this enzyme that could be used in numerous indications, e.g. hypertension. Since there is no structural information available for this important enzyme, we built a model of the complete ectodomain using the recently solved structure of human NEP as template. The naturally derived metalloproteinase inhibitor phosphoramidon was docked in the active site of this model and comparisons with the respective NEP complex were made. PMID- 11438757 TI - Comparing the effect on protein stability of methionine oxidation versus mutagenesis: steps toward engineering oxidative resistance in proteins. AB - The biological activity of some proteins is known to be sensitive to oxidative damage caused by a variety of oxidants. The model protein staphylococcal nuclease was used to explore the effect on protein structural stability of oxidizing methionine to the sulfoxide form. These effects were compared with the effects of substituting methionines with isoleucine and leucine, a potential strategy for stabilizing proteins against oxidative damage. Wild-type nuclease and various mutants were oxidized with hydrogen peroxide. Stabilities of both oxidized and unoxidized proteins were determined by guanidine hydrochloride denaturation. Oxidation destabilized the wild-type protein by over 4 kcal/mol. This large loss of stability supports the idea that in some cases loss of biological activity is linked to disruption of the protein native state. Comparison of mutant protein's stability losses upon oxidation showed that methionines 65 and 98 had a much greater destabilizing effect when oxidized than methionines 26 or 32. While substitution of methionine 98 carried as great an energetic penalty as oxidation, substitution at position 65 was less disruptive than oxidation. Thus a simple substitution mutagenesis strategy to protect a protein against oxidative destabilization is practical for some methionine residues. PMID- 11438758 TI - The role of Asn14 in the stability and conformation of the reactive-site loop of winged bean chymotrypsin inhibitor: crystal structures of two point mutants Asn14 ->Lys and Asn14-->Asp. AB - A double-headed chymotrypsin inhibitor, WCI, from winged bean seeds was cloned for structural and biochemical studies. The inhibitor was subjected to two point mutations at a conserved position, Asn14. This residue, known to have a pivotal role in stabilizing the first reactive-site loop (Gln63-Phe68) of the inhibitor, is highly conserved in the sequences of the other members of Kunitz (STI) family as well as in the sequences of Kazal family of serine protease inhibitors. The mutants, N14K and N14D, were subjected to biochemical assay and their characteristics were compared with those of the recombinant inhibitor (rWCI). Crystallographic studies of the recombinant and the mutant proteins are discussed. These studies were primarily aimed at understanding the importance of the protein scaffolding towards the conformational rigidity of the reactive-site loop. Our analysis reveals that, as the Lys14 side chain takes an unusual fold in N14K and the Asp14 side chain in N14D interacts with the loop residues by water mediated hydrogen bonds, the canonical conformation of the loop has remained effectively intact in both the mutant structures. However, minor alterations such as a 2-fold increase in the inhibitory affinity towards the cognate enzyme were observed. PMID- 11438759 TI - Activation of the low oxygen affinity-inducing potential of the Asn108(beta)- >Lys mutation of Hb-Presbyterian on intramolecular alpha alpha-fumaryl cross bridging. AB - The Asn108 beta-->Lys mutation in hemoglobin (HbPresbyterian mutation) endows a low O(2) affinity-inducing propensity to the protein. Introduction of a fumaryl cross-bridge between its two alpha 99 lysine residues also induces a low O(2) affinity into HbA. We have now engineered an alpha alpha-fumaryl cross-bridge into Hb-Presbyterian to determine the synergy or additivity, if any, that can be achieved between these two low O(2) affinity-inducing structural perturbations. Despite the presence of the additional epsilon-amino group of Lys108(beta) within the central cavity, the epsilon-amino group of Lys99(alpha alpha) of deoxy Hb Presbyterian retained high selectivity for alpha alpha-fumaryl cross-bridging, with an overall efficiency comparable to that with HbA. The alpha alpha-fumaryl cross-linking of Hb-Presbyterian reduced its O(2) affinity much more significantly than that observed with HbA, indicating a synergy between the two low O(2) affinity-inducing structural perturbations. Apparently, the alpha alpha fumaryl cross-bridge in Hb-Presbyterian activates part of the latent low O(2) affinity-inducing potential of Lys108(beta) that is generally activated in the presence of chloride. The synergy between the Asn108(beta)-->Lys mutation and the alpha alpha-fumaryl cross-bridging was conserved in the presence of chloride, but not in the presence of DPG. Furthermore, in the presence of chloride and DPG, alpha alpha-fumaryl Hb-Presbyterian accessed a low O(2) affinity T-state that is accessed by HbA, alpha alpha-HbA and Hb-Presbyterian only in the presence of IHP. Isoelectric focusing analysis suggested that the alpha alpha-fumaryl cross linking of Hb-Presbyterian induces changes in the ionization behavior of one or more of the functional groups neighboring Lys99(alpha) and Lys108(beta) [presumably His103(alpha) and/or Glu101(beta)] to compensate for the extra positive charge of Lys108(beta). Molecular modeling studies identified two potential chloride binding sites per alpha beta dimer within the middle of the central cavity of alphaalpha-fumaryl HbA involving residues His103(alpha), Arg104(beta) and Asn108(beta). The affinity of these sites is increased in alpha alpha-fumaryl Hb-Presbyterian as a result of the Asn108(beta)-->Lys mutation. Thus, the results of the present study suggest that the enhanced neutralization of the positive charges in the middle of the central cavity of Hb achieved by these two electrostatic modifications, one (the alpha alpha-fumaryl cross-bridge) acting directly and the other (the Presbyterian mutation) acting indirectly through the mediation of chloride ion binding, facilitates the alpha alpha- fumaryl-Hb Presbyterian to access a low O(2) affinity T-state structure much more readily than either Hb-Presbyterian or alpha alpha-fumaryl HbA. PMID- 11438760 TI - Investigation of the 'switch-epitope' concept with random peptide libraries displayed as thioredoxin loop fusions. AB - The 'FLITRX' random peptide library, consisting of dodecamer loop peptides displayed on a thioredoxin-flagellin scaffold on Escherichia coli, was used to select peptide sequences with affinity for a monoclonal antibody. These peptides were further screened for pH- and metal-sensitive antibody binding. Several zinc sensitive peptides were identified, termed 'switch epitopes'. A soluble, monomeric thioredoxin loop ('Trxloop') insertion analog of a FLITRX switch epitope was constructed and its antibody binding properties were characterized by Western blots. Zinc-dependent antibody recognition was maintained in the Trxloop protein although the apparent antibody affinity was lower. This Trxloop protein bound to an immobilized metal affinity chromatography matrix, similar to a 'histidine-patch' thioredoxin variant, and was reversibly precipitated by 1 mM Zn(2+) or Cu(2+) ions. Residues important for zinc and antibody binding were determined by site-directed mutagenesis. The Trxloop antibody affinity was increased by saturation mutagenesis. Biotinylated Trxloop ('Biotrxloop') variants of the original and improved affinity Trxloop proteins were constructed and characterized by surface plasmon resonance measurements. Increased antibody affinity was partially due to a slower antibody desorption rate, although the relative adsorption rates were dependent on the amount of immobilized Biotrxloop protein, indicating an influence of avidity on the apparent affinity. PMID- 11438761 TI - New secretory strategies for Kluyveromyces lactis beta-galactosidase. AB - We examined several strategies for the secretion of Kluyveromyces lactis beta galactosidase into the culture medium, in order to facilitate the downstream processing and purification of this intracellular enzyme of great industrial interest. We constructed plasmids by fusing the LAC4 gene or engineered variants to the secretion signal of the K.lactis killer toxin or to the secretion signal of the Saccharomyces cerevisiae alpha-factor. With these plasmids we transformed strains of the yeasts K.lactis and S.cerevisiae, respectively and tested beta galactosidase extracellular activity in different culture media. We achieved partial secretion of beta-galactosidase in the culture medium since the high molecular weight and oligomeric nature of the enzyme, among other factors, preclude full secretion. The percentage of secretion was improved by directed mutagenesis of the N-terminus of the protein. We developed several deletion mutants which helped us to propose structure-function relationships by comparison with the available data on the homologous Escherichia coli beta-galactosidase. The influence of the culture conditions on heterologous beta-galactosidase secretion was also studied. PMID- 11438762 TI - Sensory disturbances after focal extirpations of the human "motor" cortex. AB - From the late 1800s until approximately the middle of the 20th century, neurosurgeons made discrete motor cortex lesions in humans in attempts to reduce or eliminate a variety of involuntary movements, resulting mainly from epilepsy. In some cases, the neurosurgeons tested and recorded their patients' ability to perform various movements and to perceive various types of sensory stimuli after the operation. Although these studies have been largely forgotten, they have an immense advantage over primate lesion studies for understanding the function of the motor cortex because the patients were able to attempt to perform complex movements upon request, and to describe their perceptions of cutaneous stimuli, including integrated sensations (e.g., recognition of objects by palpation alone). We provide here a table containing the results of these studies pertaining to sensory deficits. The most consistent and persistent sensory deficits reported relate to object recognition and position sense. This finding is in keeping with recent electrophysiological studies in primates. Our analysis suggests that the "motor" cortex serves important sensory functions; hence, the term sensorimotor cortex, remains appropriate for the primate precentral (and postcentral) cortex. PMID- 11438763 TI - Effects of changing gravity on anticipatory grip force control during point-to point movements of a hand-held object. AB - We investigated the quality of predictive grip force control during gravity changes induced by parabolic flight maneuvers. During these maneuvers gravity varied: There were 2 periods of hypergravity, in which terrestrial gravity nearly doubled, and a 20-s period of microgravity, during which a manipulated object was virtually weightless. We determined grip and load forces during vertical point-to point movements of an instrumented object. Point-to-point movements were a combination of static (stationary holding) and dynamic (continuous movements) task conditions, which were separately analyzed in our previous studies. Analysis of the produced grip forces revealed that grip adjustments were closely linked to load force fluctuations under each gravity condition. In particular, grip force maxima coincided closely in time with load force peaks, although these occurred at different phases of the movement depending on the gravity level. However, quantitative analysis of the ratio of maximum grip force to the corresponding load force peak revealed an increased force ratio during microgravity when compared to that during normal and hypergravity. We hypothesize that the impaired precision of force coupling with respect to force magnitude during microgravity results from reduced feedback information about the object's mass during the stationary holding of the object in between each movement. The results indicate that the temporal grip force regulation is highly automatized and stable, whereas economical planning of force magnitude is more flexible and might reflect changes of the external loading condition. PMID- 11438764 TI - Stability boundaries and lateral postural control in parkinson's disease. AB - Postural instability is a major problem in patients with Parkinson's disease (PD). We examined balance control in PD by using center of pressure (CP) variability and time-to-contact to investigate boundary relevant postural control behavior under quiet stance leaning conditions. Postural orientation was manipulated by having patients (n = 10) and healthy older controls (n = 7) lean forward and backward with varying degrees of lean on a force platform. The subjects were instructed to lean forward or backward (either halfway or as far as possible) without bending their hips or lifting their heels or toes off the ground. Time-to-contact of the CP with the geometric stability boundary defined by the feet as well as CP position and variability were analyzed. Medio-lateral CP variability was increased in the patients with PD. Medio-lateral average time to-contact was decreased in the patients but not so in the anterior-posterior direction. In contrast to the CP variability, the medio-lateral variability of time-to-contact was lower in the patients. Patients as well as healthy older controls responded to lean manipulations with an increase in CP variability. Boundary relevant CP measures thus show clear changes in control strategies and confirm the role of lateral instability in PD. PMID- 11438765 TI - Corticospinal control of human erector spinae muscles. AB - We used transcranial magnetic stimulation (TMS) to study corticospinal excitability to erector spinae (ES) muscles during graded voluntary contractions in bilateral trunk extension (BTE) and forced expiratory breath holding (FEBH) in normal individuals. Motor evoked potentials (MEPs) could be produced in all subjects in the absence of voluntary activation. At maximum voluntary contraction, levels of surface electromyographic (EMG) activity were 4 times greater during BTE than FEBH. When EMG was normalized to maximum, MEP amplitudes increased in proportion to contraction in both tasks. MEPs in FEBH were compared with extrapolated values at similar EMG levels in BTE and were found to be larger. EMG and MEPs in left and right ES were symmetrical throughout the range of contractions in both tasks. ES muscles have a facilitation pattern similar to that previously shown in leg muscles, but subtle differences at low levels of EMG suggest that the facilitation is dependent on the task. PMID- 11438767 TI - [Ion channel dysfunction and peripheral nerve disorders]. PMID- 11438768 TI - [Disorders of awakening. Second part: secondary disorders]. AB - Secondary disorders of awakening should be distinguished from primary disorders, narcolepsy, idiopathic hypersomnia, recurrent hypersomnia, the causes of which are still unknown despite regular progress in the knowledge of the pathophysiology of narcolepsy. By definition secondary disorders of awakening are due to clearly identified causes of various origins. Two main types of secondary disorders of awakening have been distinguished: those depending on more or less voluntary sleep curtailment or on psychotropic or non psychotropic medications and those consecutive to different disorders, respiratory, neurologic, traumatic, psychotropic, infectious, metabolic, endocrinologic, and insomnia. Some of these disorders, frequent or very frequent, are polysomnographically investigated, night and day, enabling to assess in each case the type and severity of sleepiness. Others are only clinically evaluated. Disorders of awakening secondary to neurologic conditions and to a lesser extent to infectious conditions offer a special opportunity to study the anatomical basis of these disorders. They are granted more space. PMID- 11438769 TI - [Role of rehabilitation in hemineglect syndromes]. AB - Unilateral neglect syndrome compromises functional outcome of left hemiparetic patients. Treatment of this syndrome is a challenge in motor rehabilitation in order to reduce the incapacity and improve the prognosis. In this study, we present a review of the literature focusing on the different rehabilitation techniques proposed and their theoretical basis: improvement of the ability to maintain attention, improvement of spatial selective attention by implicit or explicit cueing, reduction of rightward orienting bias by sensory manipulations. Recently, we reported the improvement of symptoms of unilateral neglect syndrome after a prism adaptation to a rightward optical deviation. Unlike previous physiological manipulations used to improve neglect, this improvement lasted for at least two hours after prism removal. These results are most promising for rehabilitation programs. They may be linked to a stimulation of a short-term plasticity of brain functions and underline the interest of action in rehabilitation of neglect. PMID- 11438770 TI - [Social and economic cost of L-Dopa-induced dyskinesias in patients with Parkinson's disease]. AB - Long term L-Dopa medication mainly induces two complications in Parkinson's Disease (PD) patients, i.e. motor fluctuations and dyskinesias. Both of these complications can significantly impair the patient's quality of life. At a high degree of severity, dyskinesias can interfere with everyday tasks, balance, speech, etc. From a clinical point of view, they lead to complicated L-Dopa dose adjustments or to the use of sustained release forms. For these reasons, L-Dopa induced dyskinesias may have a significant human and financial impact. We have designed a retrospective study to explore two aspects of PD, i.e. the impact of dyskinesias on patient's social life and their economic cost. Ten French neurologists were randomly selected from a national file to participate in this study. They were asked to include 4 patients with PD currently under L-Dopa medication and showing 4 different levels of dyskinesia severity, i.e. absent, moderate, mild and severe dyskinesia, according to the Unified Parkinson Disease Rating Scale, with no other specific inclusion criteria. Two specific scales were to be completed, Hoehn & Yahr on disease severity and Schwab & England on degree of independence. Clinical profiles of patients, demographics and use of medical resources were also recorded. Investigators were interviewed about their practice. This study showed that patients with dyskinesias had an earlier onset of the disease (11.2 years vs. 3.7 years). They also had a higher level of disease severity, according to the Hoehn & Yahr scale (40p. cent vs. 0p. cent presenting a level of 4). The mean monthly cost was significantly higher for patients with dyskinesias (FF 3 733 versus FF 1 109, p=0.0005). Particularly, the presence of dyskinesias had a significant impact on treatment costs and medical visits. A positive gradient of the medical cost with level of dyskinesia severity was also underlined. In the same way, the more severe the dyskinesia status the greater the need for caregivers. Finally, based on these results, the total annual medical cost of dyskinesias in France was estimated between 588 and 812 million francs. PMID- 11438771 TI - Chronic idiopathic sensory ataxic neuropathy: immunological aspects of a series of 17 patients. AB - Sensory ataxic neuropathies (SANs) are characterized by loss of proprioceptive sensations and preservation of muscle strength. They may be idiopathic or associated with different toxic, infectious or autoimmune causes. Reactivity against gangliosides containing disialosyl groups, particularly GD1b, has been reported in isolated cases of acute and chronic idiopathic ataxic neuropathies (iSAN) and different experimental findings (in vivo animal models and in vitro preparations) suggest that antidisialosyl or antiGD1b antibodies could play a role in the pathogenesis of some ataxic neuropathies. We present the clinical, immunological and immunohistochemical characteristics of 17 patients who had a chronic iSAN without gammopathy. Patients were selected from a large group of 130 subjects with SAN: 93 with known etiology and 37 with iSAN. IgM and IgG antibodies to GM1, GM2, GM3, aGM1, GD1a, GD1b, GD3, GT1b and GQ1b were investigated by ELISA (INCAT protocol) and thin layer chromatography. Immunohistochemistry, using biotinylated Ig extracted from the patientś serum, was performed on human dorsal root ganglia (DRG), spinal cord, anterior and posterior roots, sural nerve and muscle tissue. The mean age of the 17 patients was 62 (37-80 years). The most disabling features were unsteadiness and severe ataxia of gait. Only one patient of this group was wheelchair-bound. The clinical data of these 17 patients were similar to those of the other patients with SAN, except that progression was slower. Antibodies to GD1b, GD3 and GT1b were found in 1/17. Two more patients (one with an acute iSAN and one with chronic iSAN and gammopathy), also had antibodies to disialosyl or GD1b. No immunohistochemical pattern of reactivity was found in any of the tissues tested with the 17 sera. In summary, this study demonstrates antidisialosyl or anti GD1b antibodies only in 3/37 (8.1p. cent) of patients with iSAN, either acute, chronic, or with gammopathy. However, their value seems to be reinforced by the negativity of antiganglioside antibodies in the large group of patients with SAN of known etiology (0/93). Further studies will be necessary to confirm the importance of target antigens containing disialosyl moieties in a subset of iSAN patients. However, the negativity of antiganglioside antibodies in most cases suggests that the pathology of the sensory neurons and/or axons is probably not humorally mediated in the majority of patients with iSAN. PMID- 11438772 TI - [Optimal use of lamotrigine in clinical practice: results of an open multicenter trial in refractory epilepsy]. AB - The study was designed to evaluate optimal use of add-on lamotrigine in the treatment of children and adults with refractory epilepsy of any type. Because of the available evidence from controlled studies, indicating the large spectrum of action of lamotrigine, we designed this prospective study to investigate the efficacy and safety of lamotrigine in everyday clinical practice, to collect useful information on its action in specific epilepsy syndromes and on the clinical results of specific co-medications. We studied 566 patients with a diagnosis of refractory epilepsy of any type currently receiving stable conventional regimens of antiepileptic therapy. Efficacy analysis was limited to 510 patients (388 patients aged 12 years or more, 122 patients aged 2 to 12 years) for which the exact number of seizures could be evaluated. Seizure characteristics were: simple and/or complex partial seizures in 298 (58p. cent) patients, partial seizures with secondary generalisation in 85 (17p. cent), generalised seizures of any type in 226 (44 percent). Syndromic diagnosis was partial symptomatic or cryptogenic epilepsy in 302 patients (59 percent), generalised symptomatic or cryptogenic epilepsy in 116 (23 percent) and idiopathic generalised epilepsy in 50 (10 percent). The percentage of patients who achieved at least 50 percent reduction in the frequency of seizures was evaluated around 40p. cent for all epilepsy categories, and up to 61 percent in idiopathic generalised epilepsies. Response to treatment with lamotrigine was usually obtained by the end of titration (4 weeks) and remained stable at 48 weeks. Thirty-three patients (7 percent) remained seizure-free at 48 weeks. In the group of patients with partial epilepsy, 19p. cent presented a more than 75 percent reduction in seizure frequency. A more than 50 percent reduction in secondary generalisation of partial seizures was observed in 45 percent of the patients. Efficacy results were similar in both the adult and paediatric age groups. They were better for patients receiving valproate co-medication (45 percent of the responders) as compared to other co-medications (37 percent of the responders), suggesting a synergistic action. Safety has been evaluated for all the patients having received lamotrigine (n=566). The incidence of adverse events attributed to lamotrigine was similar to the results of controlled studies, with somnolence reported in 10p. cent, a cutaneous reaction in 8 percent and episodes of transitory diplopia in 8 percent. A cutaneous reaction was more frequent in patients receiving carbamazepine (10 percent) as compared to valproate comedication (5 percent). However, the adverse event was sufficiently serious to necessitate hospitalisation in 3 patients receiving valproate. Dose escalation was not respected in two. Rash was reversible in all of the patients following discontinuation of the drug. The results of this study contribute to the overall understanding of the spectrum of lamotrigine effectiveness across seizure types and epileptic syndromes. Lamotrigine was well tolerated in children and adults. PMID- 11438773 TI - [Multiple sclerosis and Leber's hereditary optic neuropathy mitochondrial DNA mutations]. AB - Multiple sclerosis (MS) has long been known to be associated with Leber's hereditary optic neuropathy (LHON), a disease caused by mitochondrial DNA (mtDNA) mutations. We have investigated the possible involvement of LHON mtDNA point mutations in MS. The study covered a group of 75 unrelated Caucasian patients, with the relapse-remitting or primary progressive form of MS, and a control group of 75 volunteers (matched for age, gender and ethnic origin). Mitochondrial DNA from each subject was examined for 4 primary LHON mutations (at nucleotide positions 3460, 4160, 11778 and 14484) and 7 secondary LHON mutations (at nucleotide positions 4216, 4917, 5244, 7444, 13708, 15257 and 15812) by means of restriction site polymorphism and sequencing techniques. None of the primary LHON mutations were detected in the MS patients or in the controls, whereas the proportion of individuals with secondary LHON mutations was identical (27 p. cent) in the two groups. A combination of 2 or 3 homoplasmic mutations, defining mtDNA haplogroups, was found in the majority of cases. Haplogroups J, T and X were not particularly associated with MS. The frequency of the 13708 mutation alone (haplogroup X), or associated with the 4216 mutation (haplogroup J), was somewhat higher (p=0.059) in the subgroup of MS patients with optic neuritis (ON). ON was the initial symptom in all but one of the patients with haplogroups J or X. No other correlation was found between MS phenotypes and mtDNA genotypes. Our observations confirm previous reports that neither primary nor secondary LHON mutations are involved in the development of MS. However, MS patients with haplogroups J or X appear to have a moderately higher risk of developing optic neuritis. Thus, a specific mtDNA background may be a predisposing genetic factor for optic nerve damage in MS patients. PMID- 11438774 TI - [Encephalomyelopathy associated with monoclonal gammapathy]. AB - We describe the case of a patient with a particular form of presumably immune mediated encephalomyelitis associated with a monoclonal cold agglutin gammapathy. Systematic autopsy showed predominantly demyelinating lesions of the brain and spinal cord. The lesions were assumed to be the immune-mediated consequences of the underlying hematologic condition. Similarity with certain paraneoplastic syndromes is underlined. PMID- 11438775 TI - [Sarcoidosis and progressive multifocal leukoencephalopathy]. AB - A 70-year-old woman treated for sarcoidosis complained of progressive cognitive impairment and gait disability. Magnetic resonance imaging of the brain revealed a nonenhancing lesion in T1-weighted imaging in the left parieto-occipital region and sarcoidosis of the central nervous system was evoked. However, she rapidly deteriorated with posterior and cerebellar extension of the lesions, suggesting of progressive multifocal leukoencephalopathy (PML). DNA of the JC virus (JCV) was detected in the cerebrospinal fluid (CSF) by a polymerase chain reaction. Despite antiviral therapy, she died nine months after the first neurological signs. This case illustrates the possible association between sarcoidosis and PML, and underlines the interest to detect the presence of JCV in the CSF when the diagnosis of neurosarcoidosis appeared uncertain. PMID- 11438776 TI - [Pedoncular hallucinosis: implication of the thalamic and pontine structures]. PMID- 11438777 TI - [Adress from the president for 2001]. PMID- 11438778 TI - [Adress from the president for 2000]. PMID- 11438779 TI - [Can the EMG provide objective evidence distinguishing between a neurogenic and myogenic diagnosis?]. PMID- 11438780 TI - [Mini-mental state examination]. PMID- 11438781 TI - [Screening for bronchogenic carcinoma using computed tomography. More questions than answers]. PMID- 11438782 TI - [The radiologic report and its readability]. AB - AIM: The aim of this study was to evaluate the readability of radiologic reports and to determine whether they can be improved by modifying then according to precisely defined rules. MATERIAL AND METHODS: Forty reports, 10 for each diagnostic procedure (conventional radiography, US, CT, MRI); were randomly selected from a corpus of 400. The reports were analysed quantitatively using a dedicated software and qualitatively taking into account the formal, syntactic and lexical aspects of linguistically correct language and specially defined rules. On the basis of the collected results the reports were modified and analysed. RESULTS: Once again the modifications resulted in increased legibility (as testified by readability indices) that was more evident in US and X-ray reports and less evident in CT and MR reports because of the quantity of technical terms. It hoped that a radiological lexicon and guidelines for report writing will be widely adapted, so that radiologic reports will be more readable and easy understand. PMID- 11438783 TI - [Mammographic screening or clinical diagnosis? Proposal of a unified model]. PMID- 11438784 TI - [Application of a computer-aided detection (CAD) system to digitalized mammograms for identifying microcalcifications]. AB - PURPOSE: It is estimated that during mammographic screening programs radiologists fail to detect approximately 25% of breast cancers visible on retrospective review; this percentage rises to 50% if minimal signs are considered. Independent double reading is now strongly recommended as it allows to reduce the rate of false negative examinations by 5-15%. Recent technological progress has allowed to develop a number of computer-aided detection (CAD) systems. The aim of CAD is to help radiologists interpret lesions by serving as a second reader. In this study the authors developed and applied a CAD system to measure its ability to microcalcifications detect and compare its performance with that of a human observer. MATERIAL AND METHODS: The study was performed as part of the CALMA (computer-aided library for mammography) project of the Pisa section of the National Institute of Nuclear Physics. The aim of this project is to set up a large database of digital mammograms and to develop a CAD system. Our study series consisted of 802 mammograms - corresponding to 213 patients - digitalized between March and June 2000. We performed traditional mammography and then digitalized the mammograms using a CCD linear scanner (pixel size of 85 x 85 microm2, 12 bits). The images were evaluated by two radiologists with similar experience and then by the CAD system. This CAD system searches for microcalcifications by using ad hoc algorithms and an artificial neural network (Sanger type). RESULTS: The number of clusters in our database was 141 corresponding to 140 images; 692 images were non pathological. The CAD system identified a variable number of clusters depending on the threshold values. The threshold value is a number over which the probability of finding a lesion is highest. With thresholds of 0.13 and 0.14 the CAD system identified 140/141 clusters (99.3%); with a threshold of 0.15 it identified 139/141 clusters (98.6%); with a threshold of 0.16, 137/141 (97.2%); with a threshold of 0.18, 133/141 (94.3%); with thresholds of 0.18 and 0.20, 130/141 (92.2%). With threshold values of 0.13, 0.14, 0.15, 0.16 and 0.17 the system's sensitivity was greater than 82%, whereas with values of 0.18 and 0.20 it was greater than 80%. The number of false positive region of interest (ROI) / image was greater with low threshold values: in particular, thresholds of 0.13 and 0.14 yielded 16 false positives /image, thresholds of 0.15 and 0.16 yielded 9 and 7 false positives/image, and both 0.18 and 0.20 only 5/image. DISCUSSION: ROC curve shows how the use of high threshold values determined a very high specificity despite very low sensitivity rates. Conversely, low threshold values allowed to have a high sensitivity and a very low specificity. The best performance of our CAD system was obtained with threshold values at 0.15 and 0.16. In fact these thresholds resulted in a high sensitivity (greater than 82%) with an acceptable number of false positives/image, 9 and 7/image, respectively. It is not yet known how radiologists can deal with large numbers of false positives in screening programmes but in our opinion the most important feature of a good CAD system is a high sensitivity. CONCLUSIONS: In the near future the use of CAD systems will be widespread and easier to apply to everyday practice above all in centers where digital mammography is performed. Mammograms could be directly shown to radiologists after the CAD system has selected the ROI and analysed the images. Thanks to its high sensitivity and despite its low specificity CAD represents a concrete aid for radiologists. PMID- 11438785 TI - [Digital stereotactic breast microbiopsy with the mammotome: study of 122 cases]. AB - PURPOSE: To evaluate the diagnostic value of vacuum-assisted biopsy (VB) with digital stereotactic Mammotome on dubious mammary lesions clinically and sonographically negative as an alternative to surgical biopsy diagnosis. MATERIAL AND METHODS: From November 1999 to November 2000, 122 biopsies were performed using a vacuum suction aspiration device with digital stereotactic-guided Mammotome (Fischer prone table) on 116 patients between the age of 34 and 74 years (median 51 yrs). Sampling was performed with an 11 gauge probe. All patients were previously subjected to mammography in which dubious lesions were evidenced without clinical and sonogram confirmation. All lesions that were obviously benign or malignant were excluded. Mammography demonstrated isolated microcalcifications, small opacities and parenchymal distortions. Microcalcifications were then subdivided into lobular, rad-shaped, granular, mixed and pulverulent, according to Tabar's classification. The median diameter of lesions was 13 mm (range 3-50). The median number of Mammotome biopsy samples was 10.8 (range 3-19). All samples were radiographed and magnified. In 76 patients, the Mammotome biopies were incisional (only a part of the lesion was removed) and in 46 the biopsy was excisional (the entire lesion was removed). The biopsy site was marked with a non-magnetic metallic clip in 83 patients. Slight bleeding was noted during the procedure in two cases. Ultrasound visualization evidenced a hematoma at the biopsy site in one patient which was successfully resolved in one month with local therapy. No noticeable pain or patient intolerance to the procedure was observed. RESULTS: Histological probe sample examination was negative in 91 patients (74.6%) and positive in 31 patients (25.4%). The benign histologies included adenosis and epitheliosis (47 pts), sclerosing adenosis (7 pts), fibroadenoma (7 pts), fibrosis and fibrocystic mastopathy (30 pts). Of the 31 positive patients, 14 were infiltrating carcinomas (45%) and 17 ductal carcinomas in situ (DCIS) (55%). All positive cases underwent surgery. The post-surgical histological examination confirmed all diagnoses of infiltrating carcinoma while the diagnosis of DCIS was confirmed in 15 cases; infiltrating microfoci associated with DCIS were evidenced in two cases. CONCLUSIONS: VB with digital stereotactic Mammotome was shown to be a valid method for dubious mamographic lesions in which cytology often supplies an inadequate or false-negative responses. This method distinguishes between benign and malignant lesions rapidly without discomfort to the patient, thus reducing the number of diagnostic surgical biopsies. Moreover, in positive cases, it offers the advantage of avoiding extemporaneous intraoperative procedures and the possibility to perform biological tests on the tumor before surgery (estrogen and progesterone receptors, HER-2 neu, grading, Ki 67, p53 ), thus providing patient benefits and a distinct reduction of costs. However, this method is purely diagnostic and is not sufficient for therapeutical purposes. PMID- 11438786 TI - [Endovascular treatment of carotid stenosis]. AB - PURPOSE: To evaluate the efficacy and risks of endovascular treatment of carotid stenosis by percutaneous angioplasty and stenting, and to point out the potential indications of this technique compared to surgery. MATERIAL AND METHODS: From June 1996 to May 2000 we performed 84 endovascular treatments of epiaortic vessels in 79 patients with age ranging from 42 to 80. Sixty-eight symptomatic carotid stenosis greater than 70% were treated in 63 patients: 28 (41%) were post surgical restenosis of the carotid bifurcation after carotid endarterectomy and 40 (59%) were primitive carotid stenosis. In 14 cases (20%) there was occlusion of the contralateral internal carotid artery. All the procedures were performed in the angiographic suite, with local groin anesthesia and with femoral arterial approach. Angioplasty alone was done in 9 cases (13%) while stents were positioned in 59 cases (87%). The stenoses were pre-dilated in 57% of the procedures, and post-dilated in 61%. In 8 patients (12%) the endovascular treatment was performed during distal cerebral protection. During the follow-up period patients underwent echo-Doppler studies after 24 hours, at 6 and 12 months. RESULTS AND DISCUSSION: The stenotic tracts were successfully dilated in all the patients, with a residual stenosis less than 30% in 94% of cases. We didn't observe any complication in the 30 days post-operative period. One permanent neurological deficit (1.5 %) and 1 transient neurological deficit (1.5%) were observed in 2 patients within 1 hour after treatment. Other 2 patients complained a transient ortostatic hypotension. Combined major stroke and death rates was of 1,5% after 30 days post-operative follow-up. The presence of associated occlusion of the contralateral internal carotid and the presence of cardiological failure or pulmonary insufficiency has not influenced the outcome. No new clinical complication has been reported during the follow-up while the restenosis rate after the endovascular procedure is 1.5%. Our results with the endovascular treatment of the carotid stenosis appear not significantly different from those reported by the main surgical publications and trails about the carotid endarterectomy. In particular in our experience the complication rate is similar to surgery, probably due to our patient selection and to the evolution of the materials for endovascular therapy. CONCLUSIONS: Our experience points out an improvement of the efficacy and safety of the endovascular technique. We confirm the indication of the angioplasty and stenting in post-surgical restenosis and in patients with high surgical risk. Moreover we think that the endovascular treatment can be performed also in primitive carotid stenosis with acceptable complication risk. PMID- 11438787 TI - [Endovascular treatment for recurrent carotid stenosis]. AB - PURPOSE: Surgery of recurrent carotid stenosis (RCS) has higher complication rates than primary carotid endoarterectomy (CEA). Percutaneous transluminal angioplasty (PTA) and stent placement were evaluated retrospectively with a view to proposing then as alternative procedures for RCS. METHODS: In the last 10 years, 19 patients underwent 20 endovascular procedures for RCS at our Department. The mean interval between CEA and PTA was 21 months (range 4-96): 14 patients had PTA within 2 years, 3 patients within 2 and 3 years,and 2 after 3 years. The mean degree of stenosis was 92% (range 80-95%). PTA was performed by balloon catheters (size 4-7 mm) without using cerebral protection device; one self-expanding stent was used to treat RCS after PTA. All patients underwent physical examination and carotid color-coded Doppler sonography in autumn 1999 RESULTS: The procedure was technically successful, with residual stenosis lower than 50%, in 17 of 19 patients; 10 patients showed residual stenosis lower than 30%. Carotid PTA was stopped due to transient neurological deficit in one case. One RCS proved uncompliant even though high-pressure balloon catheters were used. The mean follow-up period in 16 patients was 37.4 months (range 3-99 months). Carotid restenosis after PTA developed in 3 patients, respectively after 29,18 and 7 months. In the last case RCS was successfully treated by stent placement (Wallstent). The primary patency rate was 81%, the secondary patency rate was 88% and the late clinical success rate was of 94%. CONCLUSIONS: In selected cases, PTA without the use of cerebral protection devices and stent placement proved to be a safe and effective alternative treatment for early RCS. When an atherosclerotic lesion is suspected surgery or endovascular treatment with cerebral protection devices are recommended. PMID- 11438788 TI - [Percutaneous treatment of uterine fibroleiomyomas: analysis of complications and quality of life after embolization]. AB - PURPOSE: To determine the rate complications and the quality of life of patients submitted to uterine artery embolization for the treatment of uterine fibroids. MATERIAL AND METHODS: Thirty-two women, aged 32-52 yrs underwent bilateral embolization of the uterine artery. The patients were affected by menorrhagia (100%) associated with: anaemia in 9 cases, intense menstrual pain in 10 and pressure symptoms in 3. The embolization procedure consisted of selective catheterization of the uterine arteries and injection of embolic particles (PVA). A questionnaire was administered to 27 patients over a period of 4-16 months to evaluate the patients' quality of life after the treatment. RESULTS: The procedure was technically successful in 30/32 patients. Twenty-nine of the embolized patients complained of heavy post-procedure pain associated with asthenia and malaise. Two patients were lost in the follow-up period. In seven patients (21.8%) different complications occur in the follow-up period. Two patients required hysterectomy: one for recurrent infections that required multiple admissions to hospital, and one as an emergency for the development of a large purulent collection in the uterus. Nineteen out of 27 completed the questionnaire. CONCLUSIONS: Our results indicate a high level of satisfaction with the treatment and a relatively low complication rate, which suggest that uterine embolization can be considered as a new promising option for uterine fibroids. However, before the treatment can be adopted in clinical practice, randomized trials are required to compare the treatment with the standard surgical procedures such as myomectomy and hysterectomy. PMID- 11438789 TI - [The role of transvaginal ultrasound and sonohysterography in the diagnosis and staging of endometrial adenocarcinoma]. AB - PURPOSE: The aim of this study is to evaluate the accuracy of sonohysterography in early diagnosis of endometrial tumor lesions and in the detection of myometrial infiltration for staging. MATERIAL AND METHODS: We performed sonohysterography as a preoperative test in 24 patients with an hystologic diagnosis of endometrial adenocarcinoma obtained by hysteroscopy and biopsy. The mean age of the patient was between 50 and 82 years. The sonohysterographic examination was performed by using 5.0 and 6.0 MHz transvaginal probes and a 5 or 7 French hysteroinjectors with inflating balloon. 19 of the 24 patients were enrolled in the study: in 2 cases the examination was not technically performable, 2 patients refused surgical treatment and 1 patient had a cervical adenocarcinoma with extension to the myometrium. In each patient we evaluated the number and the size of the lesions and the degree and the depth of myometrial infiltration. Each parameter was compared with the final histopathologic examination. RESULTS: Sonohysterography showed a single lesion in 15 patients, whereas in 4 patients it showed multiple lesions; in 1 of these patients it showed 3 lesions which were, in reality, a single lesion that infiltrated the first half of the myometrium. Myometrial infiltration was correctly evaluated by the examination in 17 of the 19 women (89.4%): 16 positive and 1 negative case. The sensitivity was 88%, the specificity 100%, the positive predictive value 100% and the negative predictive value 33%. The sonohysterography allowed to evaluate exactly the depth of myometrial invasion in 15 of the 16 cases (93.7%), in which a myometrial infiltration was suspected. With regard to this parameter the sensitivity was 85.7%, the specificity was 100%, the positive predictive value 100% and the negative predictive value 90.9%. CONCLUSIONS: Although the introduction of transvaginal ultrasonography in clinical practice allows to obtain an early diagnosis of endometrial adenocarcinoma, about half patients seems to present already at the diagnosis myometrial invasion. Moreover 50% of these patients seems to have pelvic lymphonodes and about 29% positive paraaortic lymphonodes. Currently myometrial invasion is evaluated by the extemporary frozen test and confirmed by the definitive hystologic examination. It would be helpful to have a technique able to detect and evaluate infiltration before surgery. The results of this study suggest that sonohysterography could have a role in preoperative staging. However these data need to be confirmed by further studies. PMID- 11438790 TI - [Radiographic patterns in patients after antireflux surgery]. AB - PURPOSE: To evaluate the usefulness and diagnostic accuracy of the radiologic examination of the oesophagus and stomach in the follow-up of patients who had an anti-reflux procedure with the purpose of identifying the normal radiographic pattern, possible surgical complications and the causes of failure of the surgical treatment. MATERIAL AND METHODS: From February 1999 to September 2000, we radiographically reviewed 11 patients (5 men and 6 women) age range 17-69, mean age 49, who had a surgical procedure of fundoplication for hiatal hernia and/or gastroesophageal reflux (7 Nissen procedures, 2 Belsey-Mark IV, 1 Toupet, 1Dor). Four surgical procedures were laparoscopic, seven were open. All patients were reviewed with single contrast (barium sulphate 60% weight/volume) and/or double contrast technique (effervescent powders + barium sulphate 250% weight/volume) and with the acquisition of radiographs in upright, supine, prone, oblique and lateral positions. RESULTS: 5 of the patients who had a fundoplication procedure suffered complications. In one patient the fundoplication was tight; in two patients the fundoplication had broken and migrated in the thorax; in one the fundoplication had loosened; in one patient it had looseed and migrated in the thorax. In the remaining 6 cases, the passage of the barium column through the fundoplication was normal, as well as the intra abdominal position of the fundoplication. CONCLUSIONS: The possibility to perform laparoscopic fundoplication procedure has increased the number of patients selected for surgical treatment. Successful relief of gastroesophageal reflux symptoms can be achieved in 90% of patients treated with antireflux fundoplication. Less than 10% of cases may have complications, with recurrence of gastroesophageal reflux manifestations. The radiographic examination of the esophagus and stomach still represents an effective diagnostic test in the follow up of these patients. PMID- 11438791 TI - [Thrombolytic therapy in myocardial infarction. Computerized tomography of encephalic complications]. AB - PURPOSE: To evaluate the CT findings of intracerebral hemorrhage in patients undergoing thrombolytic therapy for acute myocardial infarction and to correlate the type of intracerebral hemorrhage with clinical outcome. MATERIAL AND METHODS: We retrospectively reviewed the clinical records and CT scans of intracerebral hemorrhage on a total of 302 patients who underwent thrombolytic therapy for acute myocardial infarction at our institution from January 1996 to September 1999. In each patient we evaluated: the number, sites and size of hemorrhage, and the presence and severity of mass effect. The site of the hemorrhage was classified as intraparenchymal, intraventricular, subdural and subarachnoid. RESULTS: Six patients (2%, mean age 74, range 66-80) developed intracerebral hemorrhage. There was a total of 22 hemorrhages: 1 subdural hemorrhage, 6 subarachnoid, 11 intraparenchymal and 4 intraventricular. Excluding intraventricular hemorrhage, 14/18 hemorr-hages were located supratentorially. In five patients there was a fluid-blood level. Three patients had severe mass effect with midline shift. Symptoms presented within 24 hours from the administration of thrombolytic therapy in all patients. All the patients who died had a large hematoma with a severe mass effect and a severe midline shift at CT scan. In the remaining patients, the hematoma was of medium size and no mass effect was seen. CONCLUSIONS: The most common site of hemorrhage was supratentorial and intraparenchymal. Large volume intracerebral hemorrhage, multiple hemorrhages and mass effect with midline shift were associated with increased mortality. The most commonly observed finding was a fluid-blood level hematoma. PMID- 11438792 TI - [Optic pathway gliomas in patients with neurofibromatosis type 1: clinical neuroradiological integrated approach]. AB - PURPOSE: To evaluate a patient population affected by Neurofibromatosis type 1 (NF1) with special reference to optic pathway gliomas. MATERIAL AND METHODS: A total of 419 NF1 patients (222 men, 197 women) were retrospectively evaluated to show incidence, biological behaviour, neuroradiological and clinical presentation of optic pathway gliomas. Central Nervous System MR and ophthalmologic evaluations were reviewed. RESULTS: Thirty-two patients out of 419 (7.6%) showed the presence of optic pathway gliomas. Of these, 28 (87.2%) patients showed tumoral involvement of the optic chiasma. The right intracranial optic nerve was involved in 15 patients (46.8%), the left intracranial optic nerve in 18 (56.2%), the right extracranial optic nerve in 11 (34.3%), the left extracranial optic nerve in 10 (31.2%) and the right and left optic tracts in 6 (18.7%) patients. Geniculate body and optic radiation were not involved by tumour diffusion. Visual acuity testing was positive in 26/32 (81.2%), the fundus oculi in 11/32 (34.3%) patients and the visual field evaluation in 12/32 (37.5%) patients. CONCLUSIONS: In this study we showed an high correlation between optic pathway tumours and ophthalmologic abnormalities confirming the usefulness of an integrated clinical neuroradiological approach in patients affected by NF1. We also described the biological behaviour of these tumours in our NF1 patient population. PMID- 11438793 TI - [A case of pulmonary lymphangiomatosis: high resolution computed tomography (HRCT) evaluation and correlation with anatomo-radiological findings]. PMID- 11438794 TI - [Primary fibromatosis of the breast: a case report]. PMID- 11438795 TI - [Metaplasic carcinoma of the breast: a case report]. PMID- 11438796 TI - [Unusual case of solitary echinococcus cyst of the thyroid gland complicated with tracheal fistula]. PMID- 11438797 TI - [Acute renal failure with fungus ball obstruction. Report of 2 cases]. PMID- 11438798 TI - [Ultrasonography diagnosis of pneumoretroperitoneum in a patient treated with ERCP]. PMID- 11438799 TI - [A case of a large testicular simple cyst in a 5-month-old patient: ultrasonography findings]. PMID- 11438800 TI - Localization of pigment epithelium derived factor (PEDF) in developing and adult human ocular tissues. AB - PURPOSE: To localize pigment epithelium-derived factor (PEDF) in developing and adult human ocular tissues. METHODS: PEDF was localized in fetal and adult eyes by immunofluorescence with a polyclonal antibody (pAb) against amino acids 327 343 of PEDF, or a monoclonal antibody (mAb) against the C-terminal 155 amino acids of PEDF. Specificity of the antibodies was documented by Western blotting. PEDF mRNA was localized in adult retina by in situ hybridization. RESULTS: In developing retinas (7.4 to 21.5 fetal weeks, Fwks), pAb anti-PEDF labeled retinal pigment epithelium (RPE) granules, developing cones, some neuroblasts and many cells in the ganglion cell layer (GCL). In adult retinas, pAb anti-PEDF labeled rod and cone cytoplasm and nuclei of rods but not cones. Cells in the INL and GCL, choroid, corneal epithelium and endothelium, and ciliary body were also pAb PEDF-positive. Preadsorption of pAb anti-PEDF with the immunizing peptide blocked specific labeling in retina and other tissues, except for photoreceptor outer segments. In agreement with the immunolocalization with pAb anti-PEDF, in situ hybridization revealed PEDF mRNA in the RPE, photoreceptors, inner nuclear layer cells and ganglion cells in adult retina. In developing retinas 18 Fwks and older, and in adult retinas, mAb anti-PEDF labeled the interphotoreceptor matrix (IPM). Western blots of retina, cornea, and ciliary body/iris with pAb anti-PEDF produced several bands at about 46 kDa. With mAb anti-PEDF, retina produced one band at about 46 kDa; cornea and ciliary body/iris had several bands at about 46 kDa. CONCLUSIONS: PEDF, originally reported as a product of RPE cells, is present in photoreceptors and inner retinal cell types in developing and adult human eyes. Photoreceptors and RPE may secrete PEDF into the IPM. PMID- 11438802 TI - More than poor care can lead to legal liability. PMID- 11438803 TI - Field perimortem cesarean section. PMID- 11438804 TI - The writer's challenge. PMID- 11438805 TI - 15 years of service and still going strong. Hospital Wing of Memphis. AB - Early morning rush hour has barely begun when Wing I is dispatched to an MVC on I 55 in northern Mississippi. A high speed, chain reaction MVC has left one vehicle wrapped around a tree, and a difficult extrication is in progress. EMS crews radio the flight nurses that two motorists are critically injured and request a second aircraft. Wing II lifts off 5 minutes later as MedCom communicators ask how quickly we can do a turnaround flight for a cardiac patient. It promises to be a busy day for Hospital Wing of Memphis. PMID- 11438806 TI - Acetaminophen poisoning and the use of intravenous N-acetylcysteine. AB - Acetaminophen (N-acetyl-p-aminophenol) is the most widely used antipyretic and analgesic in the United States. Acetaminophen, a combination agent in approximately 125 medications, has been deemed safe and effective when used within the recommended dosage. In 1998 and 1999, however, 203,930 cases of acetaminophen overingestion were reported to U.S. poison centers, making it the leading pharmacologic agent associated with toxicity. PMID- 11438807 TI - Leadership strategies for a stronger industry. AB - In the air medical industry, as in all of health care, we are being forced to run programs as businesses. Therefore, we should consider some leadership principles from the business world to develop our own products, lines of service, and management strategies that parallel successful business ventures. PMID- 11438808 TI - The cost of family-oriented communication before air medical interfacility transport. AB - INTRODUCTION: Family-oriented communication with parents by transport teams eases the stress associated with transferring children to tertiary care. This study was conducted to determine the duration of family-oriented visits and whether the visit contributed significant cost to the mission. METHOD: Data collection was prospective and double-blind; questions were incorporated into another study. Subjects were infants or children requiring assisted ventilation and air transport to tertiary care. Time from completion of stabilization to departure and reasons for any delay were recorded. Cost of contact time longer than 20 minutes (total acceptable time for family visit and transfer to vehicle) was calculated at paramedic overtime at $0.82/minute and aircraft wait time at $200/hour if incurred. RESULTS: Forty-six patients were enrolled. In 16 cases (35%), time between completing stabilization and hospital departure exceeded 20 minutes, with "family visit" listed as the explanation. Nine of these visits incurred overtime, and two incurred aircraft wait costs. Total costs for providing communication visits more than 10 minutes long were $607 or approximately $13 per patient. CONCLUSION: The costs for visit time longer than 10 minutes are small compared with the documented benefits of family-oriented communication. However, transport personnel must be mindful of the potential to incur additional cost through overtime, aircraft wait time, or pilot replacement. PMID- 11438809 TI - Use of a physiologic scoring system during interhospital transport of pediatric patients. AB - OBJECTIVE: To determine the incidence of physiologic deterioration in critically ill and injured pediatric patients during interhospital transport with air and ground ambulance DESIGN: Prospective, descriptive study SETTING: All children were treated in regional hospitals and then transported to a pediatric tertiary care center. PATIENTS: Children (n = 100) with a median age of 1.4 years (range 1 week to 18 years) MAIN RESULTS: Three sets of physiologic scores were calculated: at the time of referral, on departure from the referring hospital, and arrival at the tertiary care center. The incidence of significant physiologic deterioration based on the calculated physiologic scores was 5.6% (n = 4) during ground and 3.4% (n = 1) during air ambulance transports. Critical events occurred in 15% of ground and 31% of air ambulance transports. CONCLUSION: No difference existed in the incidence of adverse events or physiologic deterioration when air ambulance transports were compared with ground ambulance transports for critically ill children by our team. The physiologic scoring system we chose is simple and easy to use for quality assurance. PMID- 11438810 TI - Changes in pCO2 during air medical transport of children with closed head injuries. AB - OBJECTIVES: Because low partial pressure of carbon dioxide (pCO2) can be associated with posttraumatic cerebral ischemia, we conducted a study to determine whether the pCO2 level in ventilated children with closed head injuries transported by a trained team to tertiary care was optimally maintained during transport and determine whether hand-bagging or mechanical ventilation resulted in more optimal pCO2 levels after transport. METHODS: We reviewed the hospital charts and transport records of all infants and children who had sustained a head injury and were transported by a specialized pediatric transport paramedic team to a single tertiary care facility during a 12-month period. All children were intubated and ventilated either mechanically or manually. Outcome measures were final pCO2 before transport and first pCO2 on arrival in the PICU. RESULTS: Twenty-nine children (age 0.6 to 16 years, mean 7.3, median 6) met the criteria. Fourteen patients were hand-bagged (HB), and 15 were mechanically ventilated (MV). Eleven patients (5 HB and 6 MV) started in the target pCO2 range of 35 to 40 mmHg. After transport, nine patients (all MV) had pCO2 within the target range (P < 0.01). Duration of transport (mean 63 minutes, range 15-200 minutes) did not contribute to the final pCO2 level. CONCLUSION: MV improves management of pCO2 during interfacility transport. HB significantly increases the incidence of suboptimal pCO2 and hence the risk of suboptimal cerebral blood flow. MV appears mandatory, and monitoring CO2 in transit (end-tidal or preferably point-of-care testing) should further reduce the likelihood of secondary complications from cerebral ischemia. PMID- 11438811 TI - Mutation in the gene encoding ferritin light polypeptide causes dominant adult onset basal ganglia disease. AB - We describe here a previously unknown, dominantly inherited, late-onset basal ganglia disease, variably presenting with extrapyramidal features similar to those of Huntington's disease (HD) or parkinsonism. We mapped the disorder, by linkage analysis, to 19q13.3, which contains the gene for ferritin light polypeptide (FTL). We found an adenine insertion at position 460-461 that is predicted to alter carboxy-terminal residues of the gene product. Brain histochemistry disclosed abnormal aggregates of ferritin and iron. Low serum ferritin levels also characterized patients. Ferritin, the main iron storage protein, is composed of 24 subunits of two types (heavy, H and light, L) which form a soluble, hollow sphere. Brain iron deposition increases normally with age, especially in the basal ganglia, and is a suspected causative factor in several neurodegenerative diseases in which it correlates with visible pathology, possibly by its involvement in toxic free-radical reactions. We found the same mutation in five apparently unrelated subjects with similar extrapyramidal symptoms. An abnormality in ferritin strongly indicates a primary function for iron in the pathogenesis of this new disease, for which we propose the name 'neuroferritinopathy'. PMID- 11438812 TI - A new formulation containing sunscreen (SPF 15) and 1% metronidazole (ROSASOL Cream) in the treatment of rosacea. AB - ROSASOL Cream is a novel topical formulation of 1% metronidazole in a vehicle containing sunscreens (SPF 15). This product has demonstrated efficacy in the treatment of inflammatory lesions, erythema, and telangiectasiae associated with rosacea. PMID- 11438813 TI - Antidepressant drugs in dermatology. AB - Antidepressant drugs can be an important component of the dermatologists' therapeutic armamentarium. When considering the use of psychotropic agents in dermatology two major factors should be considered: (1) the accurate diagnosis of the comorbid psychiatric disorder, and (2) the presence of proper indications for the use of antidepressant agents. Antidepressant drugs are used in the management of the psychiatric syndromes, which are most frequently comorbid with dermatologic disorders, i.e., Major Depressive Disorder, Obsessive-Compulsive Disorder, Body Dysmorphic Disorder, Post-traumatic Stress Disorder and Social Phobia. The antihistaminic and analgesic properties of some antidepressants such as doxepin and amitriptyline, are also of benefit in the treatment of some pruritic and neuralgic states. The specific guidelines, side effect profile, drug drug interactions, and the most current indications should always be obtained for any particular antidepressant agent before it is prescribed. PMID- 11438814 TI - Erythropoietin in stem cell transplantation. AB - Anemia is universal after allogeneic and autologous stem cell transplantation, with both increased red cell utilization and decreased production playing a role. This anemia sometimes is associated with a relative erythropoietin deficiency. In allogeneic stem cell transplantation, randomized trials have demonstrated improved erythropoiesis and a decrease in red cell transfusions in recombinant human erythropoietin (rHuEPO)-treated patients. Studies of rHuEPO in patients undergoing autologous stem cell transplants, however, have not shown a benefit. The role of rHuEPO in stem cell mobilization and treatment of delayed erythropoiesis has yet to be defined and further studies are needed. PMID- 11438815 TI - High-dose chemotherapy as a consolidation approach in advanced ovarian cancer: long-term results. AB - The aim of this study was to assess the long-term impact of high-dose chemotherapy (HDC) as consolidation in a large series (n = 55) of advanced chemosensitive ovarian cancer patients who were optimally cytoreduced at time of first surgery or at interval debulking surgery (IDS). HDC consisted of carboplatin (600 mg/m(2) days 1 and 2), etoposide (450 mg/m(2) days 1 and 2) and melphalan (50 mg/m(2), days 3 and 4). The primary endpoint of the study was the assessment of time to progression (TTP) and overall survival (OS). In September 2000 the overall population had a median follow-up of 55 months (range 17--137) and a TTP of 35 months with a 5-year TTP rate of 35% (CI 95%: 21--49) whereas OS averaged 75 months with a 5-year OS of 59% (CI 95%: 45--73). In patients achieving optimal primary cytoreduction the median TTP was 44 months with a 5 year rate of 43% (CI 95%: 26--60). In the same series the 5-year OS rate was 62% (CI 95%: 45--79) (median OS = 75 months). In patients who were optimally cytoreduced at the time of IDS the median TTP was 25 months and the 5-year TTP rate was 22% (CI 95%: 3--41) and median OS was 46 months with a 5-year OS rate of 50% (CI 95%: 27--73). HDC with hematopoietic support could represent an effective approach for the treatment of advanced optimally cytoreduced ovarian cancer patients with chemosensitive disease. Patients who underwent IDS because of unresectable tumors at the time of first surgery had the greater survival benefit from HDC. PMID- 11438816 TI - High-dose chemotherapy followed by stem cell transplantation in patients with resistant Waldenstrom's macroglobulinemia. AB - Preliminary evidence suggests that high-dose chemotherapy followed by autologous or allogeneic stem cell transplantation may be effective in some patients with resistant Waldenstrom's macroglobulinemia. During the last 10 years, seven patients with Waldenstrom's macroglobulinemia have received transplants at the MD Anderson Cancer Center, four with autologous and three with allogeneic stem cells. Four patients achieved partial remission, and three patients have remained alive for at least 2 years. Our data confirm the feasibility of high-dose therapy in patients with macroglobulinemia and support the need for prospective studies of this modality in patients with chemosensitive disease. PMID- 11438817 TI - Costs of high-dose chemotherapy and peripheral blood progenitor cell autograft for breast cancer. AB - The aim of the study was to analyze the real cost of single or tandem high-dose chemotherapy (HDC) and peripheral blood progenitor cell autologous transplant (PBPCT) in patients with breast cancer. We analyzed the costs of 40 PBPCT performed in 20 patients. Tandem transplant was planned for each patient. Resources used and direct costs were identified for each patient. The study was carried out using the hospital perspective and monetary values were reported in 1999 Euro. The mean cost of whole procedure for single transplant was 20,816.63 Euro, while the mean cost of tandem transplant was 38,770.83 Euro. The cost distribution in the two groups was similar: the most expensive phase of procedure was the supportive phase post transplant (about 60% of total cost), with the categories of cost most represented being professional fees (about 28%) and pharmacy (about 35%). Awaiting more convincing trials of the clinical advantage of HDC in breast cancer, our analytical evaluation of transplant costs for different therapeutic options, single or tandem, permits identification of the most expensive categories in order to intervene for cost savings. PMID- 11438818 TI - Impact of complete remission with intensive therapy in patients with responsive multiple myeloma. AB - Clinical outcomes were assessed in 68 consecutive patients with multiple myeloma of high or intermediate tumor mass that had responded to VAD or dexamethasone based therapy and were consolidated with early intensive therapy and autologous stem cell transplantation. Results were compared with those of 50 comparable patients who refused or were unable to receive intensive treatment for socioeconomic reasons. Following high-dose therapy, the rate of CR increased from 6 to 37%, with median survival prolonged by 10 months. Survival of 21 patients with disease converted from PR to CR (median 8.3 years) was significantly longer than that of similarly-treated patients who remained in PR (median 5.0 years). CR of myeloma represents the major surrogate marker of long survival and the primary goal of myeloablative treatment for patients in PR. Twelve of 18 patients with rapid reduction of myeloma protein (T(1/2) < 0.5 months), and myeloma protein reduction to <1.0 g/dl after primary therapy achieved CR (67%), identifying pretransplant features favorable to intensive therapy. Among 35 patients with slower reduction or higher residual myeloma protein, CR occurred in eight patients (23%) (P < 0.01), for whom other treatments should be considered. The kinetics of response to initial therapy should be considered in selecting patients more likely to achieve CR and consequent long survival after intensive treatment. PMID- 11438819 TI - Improved efficiency of remission induction facilitates autologous BMT harvesting and improves overall survival in adults with AML: 108 patients treated at a single institution. AB - A hundred and eight patients less than 60 years old with de novo acute myeloid leukemia were treated between 1982 and 1994 by protocols including final intensification with a transplant using autologous bone marrow purged by mafosfamide in first remission in the absence of an HLA-matched sibling donor available for allograft. From 1989, we attempted to improve tumor control by using high-dose anthracyclines in induction, by increasing from one to two the number of consolidation courses pre-transplant and by introducing intermediate doses of cytarabine in the first consolidation course. The CR rate was 77% (33/43) before 1989 and 90% (59/65) after 1989 (P = 0.06). Forty-five out of the 59 patients (76%) who achieved CR after 1989 could undergo bone marrow grafting in CR1 vs 16/33 (48%) before 1989 (P = 0.01). In spite of the higher proportion of patients above 50 years after 1989 (32%) toxicity was mild and an adequate graft was obtained more frequently after one collection. The principal factor relating to improvement in graft feasibility was the post-1989 modification of induction and consolidation regimens. This improvement in graft feasibility was associated with a better disease-free survival (DFS) (48 +/- 7% vs 32 +/- 8%, P = 0.04) and overall survival (OS) (53 +/- 6% vs 30 +/- 7%, P = 0.007) at 5 years. By multivariate analysis four factors were associated with overall survival (OS): karyotype, white blood cell count at diagnosis, treatment regimen and bone marrow grafting in CR1. This global approach should be prospectively compared with intensive chemotherapy. PMID- 11438820 TI - Comparison of allogeneic T cell-depleted peripheral blood stem cell and bone marrow transplantation: effect of stem cell source on short- and long-term outcome. AB - We report the results of a retrospective single-center study comparing engraftment, acute and chronic GVHD, relapse and survival in patients with malignant hematological disorders transplanted with allogeneic peripheral blood stem cells (alloPBSCT, n = 40) or bone marrow cells (alloBMT, n = 42). All transplants were T cell depleted by in vitro incubation with the Campath-1 monoclonal antibody. Primary graft failure occurred in none of the patients receiving an alloPBSCT compared with 3/42 of the recipients of an alloBMT. In addition, two patients in the alloBMT group showed no platelet engraftment. Recipients of PBSC had a more rapid recovery of neutrophils (median 14 days) compared to BM transplant recipients (median 32 days). Platelet recovery was also accelerated in PBSC recipients compared to BM recipients (11 vs 38 days). There was an increase in the incidence of grade II acute GVHD and chronic GVHD in patients after alloPBSCT (18% and 23%, respectively) compared to patients receiving alloBMT (5% and 8%, respectively). The 2-year cumulative incidence of relapse was similar in both groups (47%). At 6 months after transplantation, transplant-related mortality (TRM) was lower in PBSCT recipients than in BMT recipients. However, at a follow-up of 3 years TRM was similar in both groups. The disease-free survival rate at 3 years after transplantation did not differ between the groups (42% for PBSCT and 41% for BMT recipients). Our results indicate that T cell-depleted alloPBSCT compared to alloBMT is associated with a more rapid hematopoietic reconstitution and a decreased TRM at 6 months follow-up after transplantation. However, at a follow-up of 3 years, no sustained survival benefits were observed. PMID- 11438821 TI - Treatment of steroid-resistant acute graft-versus-host disease with anti thymocyte globulin. AB - Acute graft-versus-host disease (aGVHD) is a major cause of mortality after allogeneic stem cell transplantation. Although initial treatment with corticosteroids is effective in the majority of patients, 30--60% develop steroid resistance. Anti-thymocyte globulin (ATG) is commonly used as first-line therapy for steroid resistant (SR) aGVHD. However, data on its efficacy are limited. At two institutions we reviewed the results of treatment with ATG of 58 patients with SR aGVHD. Initial manifestations of aGVHD were treated with 2 mg/kg/day of methylprednisolone (MP). Equine ATG was administered as first-line therapy for SR aGVHD, a median of 9 days (range, 3 to 39) after initiation of MP. At the time of initiation of ATG, IBMTR severity indices B, C and D were observed in 6%, 40% and 54% of patients, respectively. Improvement was observed in 30% of patients treated with ATG. Skin disease was more likely to improve with ATG (79%), while progression of gut and liver aGVHD was observed in 40% and 66% of patients, respectively. Despite initial improvement, 52 patients (90%) died a median of 40 days after ATG therapy from progressive aGVHD and/or infection (74%), ARDS (15%), or relapse (11%). Only six patients (10%), three of whom had aGVHD limited to the skin at the time ATG was administered, are long-term survivors. We conclude that initial improvement of SR aGVHD occurs with ATG in a minority of patients, and very few patients become long-term survivors. Furthermore, this treatment is associated with a high rate of major complications. PMID- 11438822 TI - Inverse relationship between human herpesvirus-6 and -7 detection after allogeneic and autologous stem cell transplantation. AB - Human herpesvirus-6 (HHV-6) and -7 were analyzed in 25 and 18 patients with allogeneic (allo) and autologous (auto) stem cell transplantation (SCT), respectively, by weekly examination of viral DNA in peripheral mononuclear cells using semiquantitative PCR and serologic tests up to 12 weeks after SCT. HHV-6 DNA was detected in 29.6% and 27.9% of samples after allo- and auto-SCT, respectively. The proportions of HHV-6-DNA-positive samples increased in week 3 and 4 after allo-SCT, and in week 1 to 3 after auto-SCT. The frequency of HHV-7 DNA detection, however, was higher after auto-SCT (24.7%) than allo-SCT (12.8%) (P 10(2) copies of HHV-6 DNA (/10(5) cells) on two consecutive occasions were allo-SCT recipients and three showed clinical episodes. Conversely, three of five patients with continuous reactivation of HHV-7 were auto-SCT recipients. Thus, the frequencies of HHV-6 and -7 DNA detection showed an inverse relationship comparing allo- and auto-SCT, suggesting a different mechanism may regulate HHV-6 and -7 reactivation. PMID- 11438823 TI - Respiratory syncytial virus infection in the late bone marrow transplant period: report of three cases and review. AB - Respiratory syncytial virus (RSV) infection is an important cause of respiratory mortality in immunosuppressed patients, including bone marrow transplant (BMT) recipients. The presence of lower respiratory tract infection and infection in the pre-engraftment phase of BMT is believed to confer a poor prognosis. Three patients who underwent allogeneic BMT at our institution developed RSV pneumonia over 1 year post BMT, with the underlying disease in remission. All three were hypoxic with extensive pulmonary disease at presentation. Treatment consisted of aerosolized ribavirin and intravenous immune globulin with successful clearing of viral shedding and excellent clinical outcomes. RSV infection is probably less severe in the late post-BMT period, but needs to be considered early in the differential diagnosis of pulmonary infiltrates in this patient population. PMID- 11438824 TI - Platelet-derived growth factor enhances ex vivo expansion of megakaryocytic progenitors from human cord blood. AB - Infusion of ex vivo expanded megakaryocytic (MK) progenitor cells is a strategy for shortening the duration of thrombocytopenia after haematopoietic stem cell transplantation. The cell dose after expansion has emerged as a critical factor for achieving the desired clinical outcomes. This study aimed to establish efficient conditions for the expansion of the MK lineage from enriched CD34(+) cells of umbilical cord blood and to investigate the effect of platelet-derived growth factor (PDGF) in this system. Our results demonstrated that thrombopoietin (TPO) alone produced a high proportion of CD61(+)CD41(+) cells but a low total cell count and high cell death, resulting in an inferior expansion. The addition of interleukin-1 beta (IL-1 beta), Flt-3 ligand (Flt-3L) and to a lesser extent IL-3 improved the expansion outcome. The treatment groups with three to five cytokines produced efficient expansions of CFU-MK up to 400-fold with the highest yield observed in the presence of TPO, IL-1 beta, IL-3, IL-6 and Flt-3L. CD34(+) cells were expanded by five to 22-fold. PDGF improved the expansion of all cell types with CD61(+)CD41(+) cells, CFU-MK and CD34(+) cells increased by 101%, 134% and 70%, respectively. On day 14, the CD61(+) population consisted of diploid (86.5%), tetraploid (11.8%) and polyploid (8N--32N; 1.69%) cells. Their levels were not affected by PDGF. TPO, IL-1 beta, IL-3, IL-6, Flt-3L and PDGF represented an effective cytokine combination for expanding MK progenitors while maintaining a moderate increase of CD34(+) cells. This study showed, for the first time, that PDGF enhanced the ex vivo expansion of the MK lineage, without promoting their in vitro maturation. PDGF might be a suitable growth factor to improve the ex vivo expansion of MK progenitors for clinical applications. PMID- 11438825 TI - Intracellular cytokine profile of cord and adult blood monocytes. AB - Cord blood (CB) transplantations are associated with low graft-versus-host disease (GVHD). The pathophysiology of GVHD involves interaction and activation of different cell types, as lymphocytes and monocytes, and results in a cascade of cytokine production. After antigen or mitogen stimulation, CB monocytes release lower levels of cytokines than adult blood (AB) monocytes. In this study, the detection of intracellular IL-1 beta and TNF-alpha produced by monocytes was evaluated in response to tuberculin PPD to investigate whether the reduced capacity of CB monocytes to secrete cytokines could be related to an impaired functional activity and to a particular phenotypic profile. Results showed that the percentage of CD64(+)monocytes producing intracellular IL-1 beta and TNF alpha was significantly lower in CB and that the phenotypic profile of CB monocytes producing these cytokine (CD64(+)CD14(+)) was different to that of AB monocytes (CD64(+)CD14(+), CD64(+)CD33(+) and CD64(+) CD45RO(+)). These results suggest that the lower capacity of CB monocyte populations to produce IL-1 beta and TNF-alpha might be due to a functional immaturity of CB monocytes at the cellular level as reflected by the different phenotypic profile of CB monocytes. PMID- 11438826 TI - TCR gamma delta cytotoxic T lymphocytes expressing the killer cell-inhibitory receptor p58.2 (CD158b) selectively lyse acute myeloid leukemia cells. AB - Cytotoxic T lymphocytes (CTL) are thought to play an important role in the graft versus-leukemia (GVL) response. Unfortunately, GVL reactivity is often associated with life-threatening graft-versus-host disease (GVHD). Characterization of CTL that selectively attack leukemic cells but not normal cells may lead to the development of adjuvant immunotherapy that separates GVL from GVHD. Here, we describe TCR gamma delta (V gamma 9/V delta 1) CTL, isolated from the peripheral blood of an AML patient after stem cell transplantation (SCT), that very efficiently lysed freshly isolated acute myeloid leukemia (AML) cells and AML cell lines. Interestingly, HLA-matched non-malignant hematopoietic cells were not killed. We revealed that the killer cell-inhibitory receptor (KIR) p58.2 (CD158b) specific for group 2 HLA-C molecules negatively regulates the cytotoxic effector function displayed by these TCR gamma delta CTL. First, an antibody against HLA-C enhances lysis of non-malignant cells. Secondly, stable transfection of HLA Cw*0304 into the class I-negative cell line 721.221 inhibited lysis. Finally, engagement of p58.2 by antibodies immobilized on Fc gamma R-expressing murine P815 cells inhibits CD3- and TCR gamma delta-directed lysis. Compared to non malignant hematopoietic cells, AML cells express much lower levels of MHC class I molecules making them susceptible to lysis by p58.2(+) TCR gamma delta CTL. Such KIR-regulated CTL reactivity may have a role in the GVL response without affecting normal tissues of the host and leading to GVHD. PMID- 11438827 TI - Oligoclonal expansion of CD4(+)CD28(-) T lymphocytes in recipients of allogeneic hematopoietic cell grafts and identification of the same T cell clones within both CD4(+)CD28(+) and CD4(+)CD28(-) T cell subsets. AB - Recipients of allogeneic bone marrow grafts have clonally expanded CD8(+)CD28(-) T lymphocytes during the early period after transplantation, which leads to skewing of T cell receptor (TCR) repertoires. Here, we have addressed the question of whether clonal expansion of CD28(-) T cells is also observed in CD4(+) T lymphocytes after human allogeneic hematopoietic cell transplantation. We found that the fraction of T cells lacking CD28 expression in the CD4(+) subset was increased after transplantation, and expanded CD4(+)CD28(-) T lymphocytes carrying certain TCRBV subfamilies showed limited TCR diversity. In order to further study the ontogeny of CD4(+)CD28(-) T cells, we analyzed the complementarity-determining region 3 (CDR3) of the TCR-beta chain of CD4(+)CD28(+) and CD4(+)CD28(-) cells. We identified the same T cell clones within both CD4(+)CD28(-) and CD4(+)CD28(+) T cell subsets. These results suggest that both subsets are phenotypic variants of the same T cell lineage. PMID- 11438828 TI - Successful treatment of intravascular malignant lymphomatosis with high-dose chemotherapy and autologous peripheral blood stem cell transplantation. AB - Intravascular malignant lymphocytosis (IML) is a rare systemic disease characterized by proliferation of malignant B (rarely T) lymphoid cells within the lumina of small arteries, veins, and capillaries. Diagnosis requires skin, liver, renal, meningeal, or brain biopsy, but is rarely made ante mortem. In this report, we describe a patient who had an ante mortem diagnosis of IML as a result of a skin biopsy. Autologous peripheral blood stem cell transplantation (auto PBSCT) was successfully performed after chemotherapy. The patient has survived for more than 30 months since the onset of the disease and maintains complete remission on the 450th day post PBSCT. To our knowledge, this is the first case of IML treated by auto-PBSCT. PMID- 11438829 TI - Steroid-induced hypertrophic cardiomyopathy following stem cell transplantation in a neonate: a case report. AB - We report a case of severe left ventricular outflow tract obstruction complicating steroid therapy in an infant undergoing allogeneic transplant in the first few weeks of life for treatment of Krabbe's disease. While this complication is well known to those treating premature infants, it has not been reported in the stem cell transplant setting. For young infants undergoing allogeneic transplant who require steroid therapy, cardiac monitoring after 2--3 weeks of therapy is recommended. PMID- 11438830 TI - Amiodarone and cyclophosphamide: potential for enhanced lung toxicity. AB - Antineoplastic therapy can be associated with drug-induced lung toxicity. With the increasing use of amiodarone for cardiac dysrhythmias there is an increasing possibility of its combined use with chemotherapies for various malignancies. We report a patient on long-term amiodarone who developed biopsy-proven drug-induced lung toxicity after receiving high-dose cyclophosphamide, at a time-frame much shorter than would have been predicted with cyclophosphamide alone. The potential for enhanced lung toxicity secondary to combination of amiodarone and cyclophosphamide is discussed. PMID- 11438831 TI - Improved gene transfer efficiency to primary and established human pancreatic carcinoma target cells via epidermal growth factor receptor and integrin-targeted adenoviral vectors. AB - In this study we analyzed two ways of retargeting of Ad-vectors to human pancreatic carcinoma with the aim of enhancing the gene transfer efficiency. First, we analyzed the expression of the epidermal growth factor receptor (EGFR) on primary, as well as established pancreatic carcinoma cells by flow cytometry which revealed high expression levels of EGFR on the surface of these cells. We showed that EGFR-retargeted entry pathway using a bispecific fusion protein formed by a recombinant soluble form of truncated Coxsackie and Adenovirus Receptor (sCAR) genetically fused with human EGF (sCAR-EGF) redirects them to the EGFR leading to an enhanced gene transfer efficiency to pancreatic carcinoma cells. Since flow cytometry revealed absence of CAR expression, but the presence of at least one of both alphav integrins on the pancreatic carcinoma cells, a second way of targeting was investigated using a genetically modified Ad vector which has an RGD (Arg-Gly-Asp)-containing peptide inserted into the HI-loop of the fiber knob. This RGD targeted Ad (AdlucRGD) revealed efficient CAR independent infection by allowing binding to cellular integrins resulting in a dramatic enhancement of gene transfer. These findings have direct relevance for Ad-vector based gene therapy strategies for pancreatic carcinoma. PMID- 11438832 TI - Bcl-2 expression by retrograde transport of adenoviral vectors with Cre-loxP recombination system in motor neurons of mutant SOD1 transgenic mice. AB - We investigated genes expression by retrograde axonal transport of replication defective adenoviruses carrying genes for LacZ (AdLacZ) and Bcl-2 in motor neurons of transgenic mice expressing mutant human Cu/Zn superoxide dismutase (SOD1) gene containing a substitution of alanine for glycine at position 93. We found that intramuscular injection of AdLacZ into the tongue of mutant SOD1 transgenic mice and their wild-type littermates at various ages results in high expression of the transgene and similar time course of expression in hypoglossal cranial nerve nuclei, suggesting no difference in the behavior of the transgene expression between the two groups. Subsequently, we employed a molecular switching cassette for Bcl-2 designed to express Bcl-2 by Cre-loxP recombination using adenoviral vectors, and examined the COS7 and primary neuronal cells with the mutant SOD1 gene. The overexpression of Bcl-2 in both cells and the neuronal protection against staurosporine-induced apoptosis were observed, after dual infection of adenoviral vectors with cassette for Bcl-2 (AxCALNLBcl-2) and Cre recombinase (AxCANCre). After inoculation of AxCALNLBcl-2 followed by AxCANCre into the tongue of both mutant SOD1 transgenic mice and wild-type littermates, Bcl-2 was detected in both the injection site and the hypoglossal nuclei of brainstems, suggesting that this was the result of retrograde transport of AxCALNLBcl-2 and AxCANCre and expression of Bcl-2 by Cre recombinase in the hypoglossal nuclei. This strategy for delivery of exogenous genes such as Bcl-2 will be useful for studying neuronal death/survival and introducing foreign genes into postmitotic motor neurons, and in gene therapy for motor neuron diseases such as ALS. PMID- 11438833 TI - A transcriptional feedback loop for tissue-specific expression of highly cytotoxic genes which incorporates an immunostimulatory component. AB - Transcriptional targeting of cytotoxic genes is an important way to control toxicity associated with gene transfer therapies, but supposedly, tissue-specific promoters are often either very weak and/or leaky. In addition, the phenotypic leakiness of such tissue-specific promoters is dependent upon the toxicity of the gene being used. Therefore, we devised a transcriptional feedback loop to restrict gene expression of very potent genes to melanoma cells. We screened different elements of the human tyrosinase promoter to find one which gave no detectable expression in non-melanoma cells but was active in melanoma cell lines. This weak, but highly tissue specific, element (Tyr-300) was then used as the basis for a transcriptional amplification feedback loop in which a consensus heat shock element (HSE) was cloned upstream of Tyr-300. The cytotoxic gene was cloned downstream of the HSE-Tyr-300 element along with a mutated form of the heat shock factor-1 (HSF-1) transcription factor, which no longer requires cellular stress to activate its trimerisation, nuclear localisation and transcriptional activation properties. Low levels of expression from Tyr-300 initiated expression of both the cytotoxic and the HSF-1 genes in melanoma cells. Gradual build up of HSF-1 amplified expression through binding to the HSE to give levels of cytotoxicity similar to that provided by a CMV promoter. However, no leakiness was observed in multiple non-melanoma cell lines tested. In addition to amplifying low levels of weak tissue-specific expression, the use of HSF-1 also leads to activation of endogenous stress-related genes such as hsp70. Induction of these genes, in the presence of cell killing by the cytotoxic gene, is a highly immunostimulatory event which enhances the antitumour vaccination effects of direct tumour cell destruction. Having demonstrated the compatibility of the component elements in plasmid form, we incorporated the feedback loop into a hybrid LTR-modified retroviral vector and confirmed that the system can be effective in the form of a viral vector. The format of the feedback loop described here could be exploited for any tissue type in which a highly tissue specific element can be identified but which is itself too weak to be effective therapeutically. PMID- 11438834 TI - In vivo gene delivery into ocular tissues by eye drops of poly(ethylene oxide) poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) polymeric micelles. AB - The primary objective of this study was to investigate the feasibility of using PEO-PPO-PEO non-ionic copolymeric micelles as a carrier for eye-drop gene delivery of plasmid DNA with lacZ gene in vivo. Using pyrene fluorescence probe methods, zeta potential, and dynamic light scattering test (DLS), the ability of micelle formation of these block copolymers with plasmid was studied. Gene expressions were visualized by both the quality of enzymatic color reaction using X-gal staining and by the quantification of the substrate chlorophenol red galactopyranoside (CPRG) in enucleated eyes on day 2 after gene transfer. In addition, microscopy to identify the types of cell showing uptake and expression of the transferred gene was used. We found that the block polymeric micelles were formed above 0.1% (w/v) of block copolymer with a size of 160 nm and a zeta potential of -4.4 mV. After 2 days of topically delivery three times a day, the most intense gene expression was observed on days 2 and 3. Reporter expression was detected around the iris, sclera, conjunctiva, and lateral rectus muscle of rabbit eyes and also in the intraocular tissues of nude mice upon in vivo topical application for 48 h with a DNA/polymeric micelle formulation. Furthermore, after two enhancement treatments, the transport mechanisms of the block copolymeric micelles were found through endocytosis in tissues by enhancement through the tight junction pathway. Thus, efficient and stable transfer of the functional gene could be achieved with PEO-PPO-PEO polymeric micelles through topical delivery in mice and rabbits. These in vivo experiments indicate the possible potential use of block copolymers for DNA transfer. PMID- 11438835 TI - Cell surface display of a lysosomal enzyme for extracellular gene-directed enzyme prodrug therapy. AB - Prodrug conversion is a promising approach to cytotoxic gene therapy if an efficient transfer of the generated drug to adjacent cells can be achieved. To maximize the efficacy of this strategy we sought to develop a system that is based on a human enzyme, acts extracellularly yet in close vicinity of the transduced cell and can be used with multiple prodrugs. Results obtained with a secreted version of human beta-glucuronidase suggested that this enzyme could be a suitable candidate, although a more stringent retention of the enzyme at the site of the producer cell, such as its attachment to the cell surface, would be desirable. Here, we show that the fusion of the transmembrane domain of the human PDGF receptor to a C-terminally truncated form of human beta-glucuronidase results in its surface accumulation at high steady-state levels. Using a doxorubicin prodrug, we demonstrate that this GDEPT system produces a strong bystander effect and has potent antitumor activity in vivo. PMID- 11438836 TI - Safety of a GM-CSF adjuvant-plasmid DNA malaria vaccine. AB - MuStDO 5 is a multivalent plasmid DNA vaccine for malaria comprised of five plasmid DNAs encoding five proteins from Plasmodium falciparum and one plasmid DNA encoding human GM-CSF. To evaluate the safety of MuStDO 5, a series of pre clinical studies were conducted in mice and rabbits. In pharmacology studies in mice, GM-CSF could not be detected in the serum following either intramuscular or a combined intramuscular/intradermal administration of the vaccine, but was readily detected in the muscle following intramuscular administration. In a tissue distribution study in mice, MuStDO 5 plasmid DNA was detected by PCR initially in highly vascularized tissues, while at later time-points the plasmid DNA was detected primarily at the site(s) of injection. In GLP safety studies in mice and rabbits, repeated intramuscular/intradermal administration of the MuStDO 5 vaccine was found to be safe and well tolerated without any evidence of autoimmune pathology. PMID- 11438837 TI - Identification of methylated CpG motifs as inhibitors of the immune stimulatory CpG motifs. AB - The unmethylated CpG motifs within E. coli DNA (EC) cause immune stimulation. In contrast, mammalian DNA such as calf thymus (CT) DNA had been thought to be immunologically inert. In this article, we demonstrate that CT DNA unexpectedly specifically inhibits the immune activation by EC but not that by endotoxin. This inhibitory effect was mediated in the signaling pathway activated by EC since CT DNA markedly inhibited the CpG-induced nuclear translocation of the transcription factors, NF-kappaB and AP-1. In addition, CT DNA significantly inhibited the synergistic immune activation by EC and endotoxin. The mechanism of the inhibition by CT DNA probably did not involve the inhibition of the cellular uptake of EC. Using a CpG-depleted plasmid, we demonstrated that CpG methylation played an important role in the inhibition by CT DNA. Compared with unmethylated plasmid DNA, CpG-methylated DNA inhibited the immune activation by EC to the same extent as did CT DNA. Importantly, the inhibitory effect of CT DNA was also observed in vivo. Our results suggest that methylated DNA may be applied to alleviate the unwanted immune stimulation and inflammation in systemic inflammatory response syndrome and in gene therapy with plasmid DNA. PMID- 11438838 TI - SV40-derived vectors provide effective transgene expression and inhibition of HIV 1 using constitutive, conditional,and pol III promoters. AB - Vectors based on recombinant SV40 viruses (rSV40) are highly effective in delivering transgene expression driven by constitutive promoters. We tested here whether these vectors could be used with conditional promoters and promoters using RNA polymerase III transcription, with inhibition of HIV-1 by Tat activation response (TAR) decoys as a functional measure of effective transgene delivery and activity. TAR decoys inhibit HIV-1 Tat, a trans-activator of HIV-1 transcription. Tat acts early in the viral replicative cycle and is essential for efficient viral replication. We evaluated rSV40 gene delivery using two different inhibitors of Tat. One was a dual function polyTAR gene encoding 25 sequential TAR elements (TAR(25)), plus an antisense tat, driven either by HIV-1 long terminal repeat (HIV-LTR) as a conditional promoter, or by cytomegalovirus immediate-early promoter (CMV-IEP) as a constitutive promoter. The other inhibitor was a single TAR decoy, driven by the U6 small nuclear RNA promoter (U6 P). These decoys were delivered to unselected cells in two different human T lymphocyte lines and to unstimulated primary human peripheral blood mononuclear cells (pbmc). Gene delivery was confirmed by PCR, and expression by RT-PCR. By in situ hybridization analysis, >95% of cells were transduced. These transgene constructs protected all cell types tested from HIV-1, as measured by syncytia formation and p24 antigen release. Somewhat better inhibition of HIV-1 replication was achieved with HIV-1 long terminal repeat (HIV-1 LTR) as a conditional promoter than with the constitutive CMV-IEP. The U6-P was also very effective, driving a TAR(1) transcript. Cell viability was not detectably affected by TAR decoy expression. Thus, rSV40 vectors effectively deliver HIV-1 inhibitory RNAs using either constitutive or conditional pol II promoters, or using a pol III promoter. The versatility of this gene delivery system may prove to be useful in anti-HIV-1 therapeutics. PMID- 11438839 TI - Fatigue in polio survivors. AB - OBJECTIVES: Fatigue is commonly reported among polio survivors. The aims of the present study were to examine the incidence of perceived fatigue among a sample of Norwegian polio survivors, and to examine the association between the level of fatigue and sociodemographic and health variables. MATERIALS AND METHODS: A mailed questionnaire containing, among others, Fatigue Questionnaire, Fatigue Severity Scale, sociodemographic and health variables were sent to a representative group of 312 Norwegian polio survivors. 276 subjects (88%) answered the questionnaire. RESULTS: The incidence of fatigue among the polio survivors were considerably higher than in the normative data. Physical fatigue, more than mental fatigue, represented the major problems. Polio subjects who reported severe fatigue had significantly more other diseases and health problems than the normative group. CONCLUSIONS: The diagnosis and treatment of other or related physical conditions should be given higher priority in the management of persons with late effects of poliomyelitis, as these conditions probably can be the reasons for fatigue more than poliomyelitis sequelae alone. PMID- 11438840 TI - Driven gait orthosis for improvement of locomotor training in paraplegic patients. AB - DESIGN: Single cases. OBJECTIVE: To compare the effects of manually assisted locomotor training in paraplegic patients with the automated training by a driven gait orthosis. SETTING: ParaCare, University Hospital Balgrist in Zurich, Switzerland. METHODS: Treadmill training with manual assistance and by a driven gait orthosis was applied to two spinal cord injured patients. The first patient had an incomplete lesion at C3, the second a complete lesion at C5. They were selected by convenience sample. The EMG activity of the leg muscles rectus femoris, biceps femoris, gastrocnemius medials (GM) and tibialis anterior (TA) was visually compared for the two training methods. GM and TA activity was also quantified by calculating the variation ratio between the EMG of the patients and a set of healthy subjects. RESULTS: No significant difference between the two training methods was found according to the leg muscle EMG activity. CONCLUSION: Neuronal centers in the spinal cord become activated in a similar way by the manually assisted and the automated locomotor training. With the driven gait orthosis training sessions can be prolonged and workload of therapists can be reduced, and therefore, the automated training represents an alternative to the conventional therapy. PMID- 11438841 TI - Pain and dysesthesia in patients with spinal cord injury: A postal survey. AB - STUDY DESIGN: A postal survey. OBJECTIVES: To assess the prevalence and characteristics of pain and dysesthesia in a community based sample of patients with spinal cord injury (SCI) with special focus on neuropathic pain. SETTING: Community. Western half of Denmark. METHODS: We mailed a questionnaire to all outpatients (n = 436) of the Viborg rehabilitation centre for spinal cord injury. The questionnaire contained questions regarding cause and level of spinal injury and amount of sensory and motor function below this level. The words pain and unpleasant sensations were used to describe pain (P) and dysesthesia (D) respectively. Questions included location and intensity of chronic pain or dysesthesia, degree of interference with daily activity and sleep, presence of paroxysms and evoked pain or dysesthesia, temporal aspects, alleviating and aggravating factors, McGill Pain Questionnaire (MPQ) and treatment. RESULTS: Seventy-six per cent of the patients returned the questionnaire, (230 males and 100 females). The ages ranged from 19 to 80 years (median 42.6 years) and time since spinal injury ranged from 0.5 to 39 years (median 9.3 years). The majority (> 75%) of patients had traumatic spinal cord injury. Of the respondents, 77% reported having pain or unpleasant sensations, and 67% had chronic pain or unpleasant sensations at or below lesion. Forty-eight per cent reported that P/D could be evoked by non-noxious stimulation of the skin indicating that allodynia is present in almost half of the patients. Forty-three per cent of respondents took analgesics, 7% received antidepressants or anticonvulsants. CONCLUSION: This survey suggests that pain and dysesthesia are common and serious complaints in SCI patients. Unexpectedly, only 7% of the patients were treated with drugs considered to be most effective in neuropathic pain. This emphasizes the need for a continued research and education on P/D in SCI. PMID- 11438842 TI - Forced vital capacity in two large outpatient populations with chronic spinal cord injury. AB - OBJECTIVE: To determine the expected vital capacity in persons with chronic spinal cord injury (SCI) in relation to injury level, completeness of injury, smoking and duration of injury, as an aid to diagnosis and management of respiratory complications. SETTING: A New York City veterans' hospital and a Los Angeles public rehabilitation hospital. METHODS: Case series from the two hospitals were pooled. Participants (adult outpatients with SCI of duration >1 year, not ventilator-dependent) were evaluated by conventional forced expiratory spirometry. Cross-sectional analysis was performed, using multiple regression, on the entire population and defined subgroups. The principal outcome measure was forced vital capacity (FVC). RESULTS: In the subjects with complete-motor lesions, FVC ranged from near 100% of normal predicted values in the group with low paraplegia, to less than 50% in those with high tetraplegia. Incomplete lesions mitigated FVC loss in tetraplegia. In subjects with paraplegia, longer duration of injury was associated with greater loss, and smoking-related loss was evident at older but not at younger ages, presumably due to greater pack years in older subjects. CONCLUSIONS: Vital capacity/SCI level relationships determined here may have diagnostic and prognostic value. Smoking-related FVC loss is important in persons with SCI as in others, although at higher levels it may be obscured by SCI-related loss. PMID- 11438843 TI - Fibrous adhesive entrapment of lumbosacral nerve roots as a cause of sciatica. AB - STUDY DESIGN: Report of seven patients with fibrous adhesive entrapment of lumbosacral nerve roots as a cause of sciatica, whose radiographic findings were negative and who experienced relief from sciatica immediately after the entrapment was released. OBJECTIVES: To describe a new clinical entity of fibrous adhesive entrapment of lumbosacral nerve roots with negative radiographic findings. SETTING: Orthopaedic department, Japan. METHODS: Clinical evaluation and post-operative outcome in seven patients with entrapment of lumbosacral nerve roots because of fibrous adhesion confirmed intraoperatively. RESULTS: Radiographic examinations by magnetic resonance imaging (MRI), myelography, and computed tomographic (CT) myelography demonstrated neither disc herniations nor spinal stenosis in all seven patients, and differential nerve root block was effective for relieving sciatica and low back pain. We confirmed, intraoperatively, entrapment of the nerve root by fibrous adhesion, and all seven patients were relieved from sciatica and low back pain postoperatively. CONCLUSION: This study presented seven patients with sciatica caused by fibrous adhesive entrapment of lumbosacral nerve roots who underwent decompression and release of fibrous adhesion. Radiographic examinations, such as MRI, myelography and CT myelography, showed no compressive shadows and also differential nerve root block was effective for its diagnosis. This study seems to be the first report of patients with entrapment of lumbosacral nerve roots caused by fibrous adhesion, whose radiographic findings were negative. PMID- 11438844 TI - Incidence rates and populations at risk for spinal cord injury: A regional study. AB - STUDY DESIGN: A 6 year retrospective study was conducted. OBJECTIVES: The populations at risk for spinal cord injury (SCI) in the northwestern Kentucky (KY) and southern Indiana (IN) regions were identified following examination of the causes and factors associated with SCI. SETTING: The database included patients primarily from the surrounding KY and IN counties admitted to the University of Louisville (U of L) Hospital. METHOD: Specification of SCI patient demographics, injury causes and related factors was achieved utilizing the hospital's trauma institute database and an extensive review of patient medical records. RESULTS: An adjusted average incidence rate of 27.1 cases per million per year was obtained for this region. A high rate of SCI was found for the youngest age group, 14-24 year olds, and for African Americans. A high frequency of injuries was also observed for adults between the ages of 25-39 years. Motor vehicle accidents (MVA) were the leading cause of SCI. Contributing factors included alcohol and widespread non-use of vehicle safety precautions. CONCLUSIONS: In addition to the high proportion of youth at risk for SCI, a higher proportion of older adults with SCI was observed for this region compared to other studies. Because the primary source of transportation in this area is the use of private vehicles, rather than public transportation, greater effort is warranted in emphasizing the potential risks of combining driving with alcohol consumption and non-use of seatbelts. PMID- 11438845 TI - Does a colostomy alter quality of life in patients with spinal cord injury? A controlled study. AB - STUDY DESIGN: Prospective controlled comparative analysis. OBJECTIVE: To determine whether a colostomy changes quality of life in patients with a spinal cord injury. METHOD: A previously validated questionnaire designed to assess quality of life in spinal injured patients (Burwood Questionnaire) was sent to 26 spinal cord injured patients with colostomies and 26 spinal cord injured patients without colostomy. The two groups were matched for level of injury, completeness of injury, length of time since injury, age (+/- 5 years) and gender. RESULTS: There was 100% completion of the questionnaire. There was no significant difference (P > 0.05) in the two groups of patients in regard to their general well being, emotional, social, or work functioning. CONCLUSIONS: Patients with colostomy following spinal injury are no worse off in regard to quality of life, than those without. The inference is that perhaps a colostomy should be considered earlier in patients with major bowel dysfunction following spinal cord injury. SPONSORSHIP: Financial support for Dr AC Lynch was provided by Royal Australian College of Surgeons with a Foundation Scholarship and Grant in aid by the Burwood International Spinal Trust. Mr N Randell was supported by the Canterbury Medical Research Foundation with a summer studentship. PMID- 11438846 TI - Radioisotopic control for baclofen pump catheter failure. AB - STUDY DESIGN: Case report of Baclofen pump catheter failure investigated by radioisotope injection. OBJECTIVES: To report a safe and reliable method for evaluating catheter dysfunction. SETTING: France. METHODS: Single case report of failure of Baclofen pump investigated by radioisotope injection. RESULTS: The injection demonstrated the block in the catheter. The catheter failure was not visualised by plain X-ray nor by filling the pump with radio-opaque solution. CONCLUSION: Catheter failure is a common cause of intrathecal drug delivery problems and may be difficult to diagnose. When catheter disconnection, kink, or dislodgement is not visible on X-ray, radioisotopic control is a safe and reliable method for assessment. PMID- 11438847 TI - Bilateral extracorporeal shock wave lithotripsy in a spinal cord injury patient with a cardiac pacemaker. AB - OBJECTIVES: To review the precautions to be observed before and during extracorporeal shock wave lithotripsy (ESWL) in spinal cord injury (SCI) patients with a cardiac pacemaker and the safety of bilateral ESWL performed on the same day. DESIGN: A case report of bilateral ESWL in a SCI patient with a permanent cardiac pacemaker. SETTING: The Regional Spinal Injuries Centre, Southport, the Lithotripsy Unit, the Royal Liverpool University Hospitals NHS Trust, Liverpool, and the Department of Cardiology, Manchester Royal Infirmary, Manchester, UK. SUBJECT: A 43-year-old male sustained a T-4 fracture and developed paraplegia with a sensory level at T-2. During the post-injury period, he developed episodes of asystole requiring implantation of a dual chamber (DDD) permanent pacemaker. Twenty-one months later, he developed a right ureteric calculus with hydronephrosis. A radio-opaque shadow was seen in the left kidney with no hydronephrosis. During right ureteric stenting, the ureteric stone was pushed into the renal pelvis. 1,500 shock waves were delivered to this stone on the right side, followed by ESWL to the left intra-renal stone with 1250 shock waves. RESULTS: The patient tolerated ESWL to both kidneys. The pacemaker was reprogrammed to a single chamber ventricular pacing mode at 30 beats per minute with a reduced sensitivity during lithotripsy. There were no untoward cardiac events during or after lithotripsy. The serum creatinine was 45 micromol/l before lithotripsy and 44 micromol/l two weeks after ESWL. CONCLUSION: SCI patients with a cardiac pacemaker may be able to undergo extracorporeal shock wave lithotripsy following temporary reprogramming of the pacemaker. Bilateral, simultaneous ESWL is safe in the vast majority of patients provided that there is no risk of simultaneous ureteric obstruction by stone fragments. However, it should be remembered that a decrease in renal function could occur following bilateral ESWL of renal calculi. PMID- 11438848 TI - Total anterior urethral reconstruction with the 'BAES flap' in a spinal cord injured patient. AB - STUDY DESIGN: A case report of urethral destruction in a spinal cord injured (SCI) patient. OBJECTIVES: To report the reconstruction of the whole anterior urethra in one-stage using an epilated scrotal flap. SETTING: Institut Guttmann, Hospital de Neurorehabilitacio, Barcelona, Spain. METHODS: A one-stage tubular substitution urethroplasty based on a bi-axial epilated scrotal flap design ('BAES flap') was performed successfully. RESULTS: Long-term follow-up of 6 years has confirmed the excellent adaptation of the flap to its urethral function. CONCLUSION: The bipedicle epilated scrotal flap can effectively resolve this challenging urethral pathology. PMID- 11438850 TI - Conservative treatment of the neuropathic bladder in spinal cord injured patients. AB - Different conservative treatment modalities for the lower urinary tract dysfunction in patients with spinal cord lesion are reviewed. Conservative treatment is still the mainstay of the urological management in these patients. Growing experience has changed the classical approach. Spontaneous voiding with and without triggered voiding and/or bladder expression has proven to be less safe except in well defined patients with regular urological follow-up. Nowadays, intermittent catheterisation and self catheterisation with and without bladder relaxants are accepted as the methods of choice. Condom catheters are still needed if incontinence persists, while penile clamps have no place in the treatment of patients with spinal cord lesions. Long-term indwelling catheters should be avoided. External electrical stimulation can be used to correct the neurogenic dysfunction by neuromodulation and/or to induce a direct therapeutic response in the lower urinary tract. PMID- 11438851 TI - Ageing with spinal cord injury: cross-sectional and longitudinal effects. AB - STUDY DESIGN: Longitudinal and cross-sectional. OBJECTIVE: To determine whether, for studies of ageing with a spinal cord injury, the cross-sectional differences in outcomes across both age and years post injury (YPI) differ from the longitudinal change. SETTING: Two SCI centres in England: the National Spinal Injuries Centre in Aylesbury, and the Regional Spinal Injuries Centre in Southport. METHODS: A total of 315 people who sustained spinal cord injuries prior to 1971 underwent comprehensive health and psychosocial status interviews at one or more of the study assessments (1990, 1993, 1996, and 1999). A range of continuous and dichotomous outcomes was analyzed to detect both cross-sectional differences by age and average individual changes over multiple measurements. RESULTS: Frequently, outcomes changed longitudinally without showing any cross sectional differences. Cross-sectional age was more commonly associated with the worsening of a condition while cross-sectional YPI was commonly associated with improvement. After controlling for cross-sectional effects, psychological measures generally showed minor deterioration, measures of community integration both improved and deteriorated, upper extremity pain increased, lower enxtremity pain decreased, and participants tended to quit smoking. CONCLUSION: Using longitudinal findings that control for cross-sectional differences produces a more complete description of ageing with a spinal cord injury. PMID- 11438852 TI - The relationship between neurological level of injury and symptomatic cardiovascular disease risk in the aging spinal injured. AB - STUDY OBJECTIVES: To describe the distribution of clinically apparent cardiovascular disease (CVD) in people with long-term spinal cord injury (SCI) according to neurologic level and severity of injury. DESIGN: Historical prospective study. SETTING: Two British Spinal Injuries Centers. PARTICIPANTS: Five hundred and forty-five individuals surviving at least 20 years with SCI were divided into three neurologic categories by level of injury and Frankel/ASIA grade as follows: Tetra ABC, Para ABC, and All D. MAIN OUTCOME MEASURES: Cardiovascular disease outcomes defined by ICD/9 codes 390-448 and obtained through medical record review. Cardiovascular disease outcomes measured included All CVD, coronary heart disease (CHD), hypertension, cerebrovascular disease, valvular disease, and dysrhythmia. RESULTS: After age-adjustment, the rates of All CVD were 35.2, 29.9, and 21.2 per 1000 SCI person-years in the Tetra ABC, Para ABC, and All D groups, respectively. Rates of All CVD increased with increasing age in all neurologic groups. Tetraplegic level of SCI conferred an excess 16% risk of All CVD (95% Confidence interval [CI], 0.93-1.46), a fivefold risk of cerebrovascular disease (relative risk [RR] 5.06; 95% CI, 1.21-21.15), and 70% less CHD (RR 0.30; 95% CI, 0.13-0.70) when compared with paraplegics. More complete SCI was associated with an excess 44% All CVD risk (95% CI, 1.16 1.77). CONCLUSIONS: Risk of All CVD increased with increasing age, rostral level of SCI, and severity of SCI. More rostral level of SCI was associated with cerebrovascular disease, dysrhythmia, and valvular disease. Conversely, there was an inverse relationship between level of SCI and CHD PMID- 11438853 TI - Quality of life of primary caregivers of spinal cord injury survivors living in the community: controlled study with short form-36 questionnaire. AB - OBJECTIVE: To compare the quality of life scores of primary caregivers of spinal cord injury survivors living in the community with healthy age matched-population based controls and to determine the relationship between some severity parameters related with spinal cord injury and the quality of life scores of primary caregivers. SETTING: University hospital, rehabilitation centre. METHODS: Fifty primary caregivers of spinal cord injured patients living in the community and 40 healthy age-matched controls completed SF-36 (short form-36) questionnaire forms. Questionnaires were administered by interviewers who were physiatrists and the authors of the present study. All the patients were rehabilitated by the authors and data about the duration of injury, lesion levels, ASIA scores, degree of spasticity, presence of bladder and bowel incontinence and pressure sores were gathered from the hospital recordings and/or by physical examinations during control visits when the primary caregivers were administered the questionnaires. RESULTS: Quality of life scores measured by SF-36 were significantly low in the primary caregivers group compared to age-matched healthy population based controls. No significant relation was demonstrated between the quality of life scores of primary caregivers and parameters such as the duration of injury, lesion levels, ASIA scores, degree of spasticity, bladder and/or bowel incontinence and pressure sores respectively. CONCLUSION: According to the results of the present study, being a primary caregiver of a spinal cord injured victim significantly interferes with quality of life; some severity parameters related to the injury however do not seem to have an additional impact on the primary caregiver's life quality. PMID- 11438854 TI - Re-examining the psychology of spinal cord injury: a meaning centered approach from a cultural perspective. AB - OBJECTIVES: To examine the impact of SCI from a non-Western cultural perspective. SETTING: India. METHODS: A philosophical approach. CONCLUSION: Management of the individual with spinal cord injury needs to take in to account the individual's beliefs and cultural context. PMID- 11438855 TI - Functional community ambulation requirements in incomplete spinal cord injured subjects. AB - STUDY DESIGN: A group of people with incomplete spinal cord injuries (SCI) were evaluated and compared with able-bodied individuals during several walking conditions. OBJECTIVES: To evaluate the functional community ambulation and estimated energy expenditure in persons with incomplete SCI and able-bodied individuals. METHODS: A list of criteria was used to evaluate functional community ambulation among participants. Physiological variables, such as the heart rate, oxygen uptake and the lactate concentration, were also measured. RESULTS: Three of nine incomplete SCI subjects and all able-bodied subjects were able to meet all the criteria measured. The required velocity to safely cross an intersection was the criterion that the incomplete SCI group had the most difficulty reaching. The able-bodied subjects had a comfortable walking velocity twice that of the incomplete SCI subjects' preferred velocity. When walking at the same velocity (incomplete SCI subjects' preferred velocity), the incomplete SCI subjects had a rate of oxygen uptake 26% greater than the healthy subjects and were 200% less efficient. The lactate concentration also proved to be a useful tool when evaluating the incomplete SCI subjects' walking efficiency. The incomplete SCI subjects lactate concentration increased after walking at their preferred velocity, meaning that the anaerobic pathways were used to meet energy demands. CONCLUSION: Rehabilitation centers should adapt their evaluation forms and increase their criteria requirements to more suitable criteria that are found in the SCI patient's community. The physiological cost should also be taken into consideration when evaluating the SCI patient's functional ambulation. PMID- 11438856 TI - Metabolic and cardiopulmonary responses to acute progressive resistive exercise in a person with C4 spinal cord injury. AB - STUDY DESIGN: Single-subject (female, 38 years of age) case. OBJECTIVES: To describe metabolic and cardiopulmonary responses to progressive resistive exercise in an individual with C4 ASIA A tetraplegia, and to review the relationship between level of spinal cord injury (SCI) and exercise responses. SETTING: Large, urban mid-western city rehabilitation hospital in United States of America. METHODS: Bilateral shoulder elevation/depression (shoulder shrug) exercise with two different resistances (0.7 kg/shoulder, 1.4 kg/shoulder) at two different frequencies (20 min., 40 min.), for 2 min per bout, deployed in a discontinuous protocol. RESULTS: Compared to rest heart rate (HR), exercise HR increased the greatest (13 bpm) for the 1.4 kg resistance at 40 min. and the least (6 bpm) during the 0.7 kg at 20 min. Blood pressure (BP) response was lower than resting BP for all four exercise conditions with the lowest (74/56 mmHg) at 1.4 kg at 40 min. Oxygen uptake was highest (4.6 ml.kg(-1) min(-1)) during 1.4 kg at 20 min and V(E) was greatest (18.2 L/min) during 1.4 kg at 40 min. Rate of perceived exertion (RPE) was the highest (17) during the 1.4 kg at 40 min. CONCLUSIONS: Progressive resistance exercise provoked intense perceived physical effort, but only small metabolic and cardiopulmonary increases in a person with C4 SCI. Exercise recommended at a 'somewhat hard' intensity should avoid significant hypotension and still impressively increase oxygen uptake and ventilation compared to rest. An inverse relation between level of injury and aerobic responses may extend rostrally to the C4 level. PMID- 11438857 TI - Spinal cord injury in Vietnamese combat. AB - This report deals with the experience gained by observation and treatment of 114 Vietnamese combat injuries from December 1965 to April 1969. The report covers all aspects of rehabilitation, medical, physical and socio-economic. PMID- 11438860 TI - [Cellular biology and ophthalmology]. PMID- 11438858 TI - Management of reduced urine output in the patients with acute cervical spinal cord injury. PMID- 11438861 TI - [Hypervision: myths and reality]. PMID- 11438862 TI - [Retinal apoptosis]. PMID- 11438863 TI - [Severe hyperglycemia caused by dorzolamide]. PMID- 11438864 TI - [Aqueous humor and serum interleukin-6 in patients with uveitis]. AB - PURPOSE: A study on local and systemic behavior of interleukin-6 in patients with active uveitis. METHODS: IL-6 levels were measured in aqueous humor and peripheral blood samples using an enzyme-linked immunosorbent assay (ELISA) from 23 patients with uveitis and 16 control patients who had been operated for uncomplicated cataracts. RESULTS: Aqueous humor of patients with uveitis showed higher levels of interleukin-6 than those of controls (p<0.001). A comparison of cytokine levels between aqueous humor and serum from patients with uveitis showed significantly higher levels of interleukin-6 in aqueous than serum (p<0.001). Correlation studies using regression test for successive steps failed to demonstrate any association between interleukin-6 levels and the different clinical characteristics of uveitis patients (laterality, onset, patterns, visual damage, localization, inflammatory activity, etiology, and association with the B27 + histocompatibility antigen). CONCLUSIONS: IL-6 is a cytokine that actively participates in the pathogenesis of clinical uveitis. Our data emphasize the greater local than systemic participation of this cytokine. PMID- 11438865 TI - [Ocular blood flow and characteristics of retinal detachment]. AB - PURPOSE: To determine if ocular blood flow (OBF) changes in retinal detachment (RD) are related with its characteristics (extension, macular involvement, PVR or duration). METHODS: 60 patients with unilateral rhegmatogenous RD and 20 <> patients were selected. Color Doppler Ultrasound was used to measure OBF velocities in the ophthalmic artery (OA), central retinal artery (CRA) and vein (CRV) and posterior ciliary arteries (PCA) of both eyes of each patient. The IOP and RD characteristics were considered. In each case the paired Student's T test (OBF) and Wilcoxon test (IOP) were used for the statistical analysis (p<5%). RESULTS: Peak systolic (PS) and end diastolic (ED) velocities of OA, PS of CRA and PS and ED of PCA were decreased in the RD eyes compared to the normal ones (p<0.05). The PS and ED rates of OA and PS of CRA were also decreased (p<0.05) in the other eyes compared to the normal ones. Both the decrease in velocity and in the IOP were related with 2 or more quadrants of RD (p<0.05), macula detachment (p<0.05), PVR >/=C(2) (p<0.05) and duration of >/=15 days (p<0.05). CONCLUSION: RD correlates with decreased ocular blood flow velocities and IOP in both the RD as well as in the other eye. The worse the characteristics of the RD, the lower the velocities and the IOP PMID- 11438866 TI - [Study of subretinal neovascular membranes secondary to degenerative myopia]. AB - PURPOSE: To study the type, size, location and posterior time evolution of the subretinal neovascular membranes (SRNM) secondary to degenerative myopia (DM) and its relationship with the degree of myopic ametropia. METHODS: We studied 65 patients (130 eyes) with MD who were divided into three groups according to their refraction. A complete ophthalmologic examination, including a fluorescein angiography (FAG), was performed for all the patients. Based on this test, 55 SRNA were identified, and were classified into two types according to the fluorescein angiography results. Their position was described in relationship to the foveal avascular zone (FAZ) and their size was measured. RESULTS: 55 (42.3%) of the 130 eyes presented SRNM, which was bilateral in 16.32% of the patients. Based on the FAG criteria, 41 eyes (74.54%) presented type 1 SRNM, 60% (33 eyes) presented subfoveal membranes and the size was greater than 1,500 microns in more than 75%. The development of the SRNM showed no relationship with the myopic ametropia degree (P<0.2500). The follow-up time was 24.61+/-7.63. CONCLUSIONS: No relationship between the appearance of the development of the SRNM and the myopia degree has been found. These are generally small sized, have poor dye diffusion and are frequently located in the foveal zone. The visual acuity loss is often sudden and maintained over time. PMID- 11438867 TI - [Rational management of applanation tonometry in myopia after LASIK]. AB - PURPOSE: To determine the accuracy and to identify pitfalls when intraocular pressure is measured in myopic patients after laser in situ keratomileusis (LASIK). METHODS: A retrospective study of intraocular pressure (IOP) measured with a Goldmann applanation tonometer in 103 eyes of 52 myopic patients who underwent LASIK was performed. Patients without ocular hypertension or glaucoma were selected. The same surgeon carried out a standard LASIK technique using a microkeratome Chiron Hansatome and the Technolas Chiron 217-C Lasik. IOP was measured pre-operatively, and at 1, 3 and 6 months post-operatively. RESULTS: After LASIK, a significant decrease was observed in the IOP in the post-operative controls, with a mean decrease of 2.69+/-1.69 mm Hg (p<0.001). This was related with the degree of myopia (p=0.05) and the characteristics of the excimer-laser treatment parameters, such as corneal ablation (p=0.04). No correlation was observed between this decrease and age or gender. CONCLUSIONS: Our retrospective review showed that the post-operative intraocular pressure measured after LASIK was lower than the pre-operative ones, which changed the Goldmann tomometer accuracy, thus causing it to underestimate the IOP. PMID- 11438868 TI - [Consecutive exotropia surgery]. AB - PURPOSE: To evaluate the factors involved in the development of consecutive exotropia (XTc), and the surgical procedures used for its treatment. PATIENTS AND METHODS: A retrospective study on 30 patients who underwent surgery for XTc was carried out in our Department. The following characteristics were studied prior to surgery: anamnesis, refraction, deviation angle measurement, detection of amblyopia and diplopia prior to the operation, abnormal head posture rotations and presence of any vertical deviation and anisotropy. The surgical technique used was individually considered for each patient and included bilateral lateral rectus recessions and/or unilateral medial rectus advancement to its/their original insertion site. RESULTS: Before surgery, 53.33% of our patients showed amblyopia, 66.66% of them showed rotation limitation, 46.66% showed dissociated vertical deviation (DVD), 20% had abnormal head posture and 10% diplopia. <> results (residual deviation of 10 prism diopters or less) were obtained in 70% of our patients. More than half of our cases were corrected with one single procedure. CONCLUSIONS: The presence of amblyopia, rotation limitations and vertical deviations (DVD and/or alphabetical syndromes) were found to be the most common factors in the development of a XTc in our study. The results indicate that a residual angle less than 10 prism diopters is obtained in 70% of our patients. The technique of choice is a bilateral lateral rectus muscle recession for deviations up to 35 prism diopters. The association of an advancement of one or both medial rectus muscles is necessary when the initial deviation exceeds 35 prism diopters. PMID- 11438869 TI - [Outer retinal edema or multifocal choroiditis after retinal detachment surgery. A case report]. AB - CASE REPORT: We describe a case of a woman who, in the postoperative period of retinal detachment, developed an ophthalmoscopic picture which could correspond to abnormal outer retinal swelling or multifocal choroiditis. Ophthalmoscopic appearance consists of multiple yellowish subretinal lesions in the temporal area of the posterior pole with a cobblestone appearance. DISCUSSION: The two entities are extremely rare complications of rhegmatogenous retinal detachment surgery, with very similar funduscopic appearance. The first one consists of localized pockets of subretinal fluid remaining after retinal detachment surgery. Multifocal choroiditis is an inflammatory process related to various causes. PMID- 11438870 TI - [Physiopathology and treatment of cataract]. PMID- 11438872 TI - [What have we learned from the multicenter study on optic neuritis corticosteroid treatment?]. PMID- 11438873 TI - [Advances in the treatment of optic neuritis]. PMID- 11438874 TI - [Lipids and age-related macular degeneration]. PMID- 11438875 TI - [Clinical course of cytomegalovirus retinitis with the different antiretroviral therapies in our hospital]. AB - PURPOSE: A study of protease inhibitors on CMV retinitis in HIV infected hospital patients. METHOD: A retrospective study was performed in 485 diagnosed of AIDS, between January, 1990 and December 1998. Among those, 81 patients who were diagnosed of CMV retinitis, the following was studied: anti-retroviral treatment, anti CMV treatment, survival time and CD4 lymphocyte count. RESULTS: Treatment with protease inhibitors in HIV patients showed an increase in life expectancy, reduced the incidence of CMV retinitis and a reduction in CMV retinitis prevention treatment was observed. CONCLUSIONS: Treatment with protease inhibitors has reduced the rate of CMV retinitis in HIV patients. PMID- 11438876 TI - [Retinal detachment surgery after refractive surgery]. AB - PURPOSE: To analyze the characteristics of retinal detachments (RD) in myopic patients who underwent different kinds of refractive surgery (RS). Modifications in patient eyes deriving from RS and how it hindered surgery for treating RD, was also studied. MATERIAL AND METHODS: 14 (6 female/8 male) patients with RD after having been treated with RS (laser assisted in situ keratomileusis (LASIK), photorefractive keratectomy (PRK), radial keratotomy (RK) and phakic anterior chamber lenses). RD was treated by scleral surgery and in two cases by vitrectomy after first RD surgery. RESULTS: 12 RD cases showed slight modification over previous refraction. In 2 cases vitrectomy was necessary since proliferative vitreoretinopathy was evidenced. The appearance of RD cases did not increase after PRK, RK or LASIK. RS did not hinder RD surgery. RD appearance in patients with anterior chamber lenses was higher, with severe difficulty in peripheral exploration and in performing RD surgery. CONCLUSION: After PRK, RK and LASIK, RD surgery does not show more difficulty, while after anterior chamber intraocular lenses implantation there is an increased difficulty for RD surgery. PMID- 11438877 TI - [Application of confocal microscopy in the study of the relationships between astrocytes and blood vessels in the retina]. AB - OBJECTIVE: To study the rabbit retina astrocytes by using immunofluorescence techniques together with the confocal microscopy. MATERIAL AND METHODS: The rabbit retinas were processed with an anti-glial fibrillary acidic protein (anti GFAP) for astrocyte detection and with propidium lobide for nuclear staining. RESULTS: Confocal microscopy allows for three-dimensional reconstruction of astroglial cells, the performance of double staining procedures with superposition of images corresponding to each chromogen, the exchange of observation axes for each slide and finally the performance of serial optic sections that indicate the exact cell location and their relationship with adjacent structures, eliminating the background signals. CONCLUSIONS: The confocal microscopy provides detailed information about tridimensional morphology and the location of the astrocytes in the rabbit retina. The astrocytes associated with the nerve fiber bundles are located in the nerve fiber layer. The type III perivascular astrocytes are located between the intravitreous capillaries close to the internal limiting membrane of the retina. The type I perivascular astrocytes are found in the retinal face of the intravitreal capillaries, these being the neurons which are the most distant from the retina. PMID- 11438878 TI - [Histopathological and immunohistochemical study of the ocular cicatricial pemphigoid]. AB - OBJECTIVE: To study the relationship between the histopathological alterations and the clinical severity grade in the first phases of the ocular cicatricial pemphigoid to facilitate early diagnosis of the disease and initiate treatment as early as possible. MATERIAL AND METHODS: We have studied 10 patients with different clinical grades of severity (3 patients with grade I; 4 patients with grade II; 2 patients with grade III, and 1 patient with grade IV). For these patients, full ophthalmologic examinations were performed [clinical, laboratory tests, impression cytology, ultrastructural study, study of direct (IFd) and indirect immunofluorescence (IFi). RESULTS: The different techniques used have enabled us to observe the diminution of goblet cells, the initiation of subepithelial bullae, separation of epithelial basement membrane, increase and disorganization of collagen fibers, increase in the number of desmosomes, decrease in the number of hemidesmosones, increase in the intraepithelial fibrillar materials and the presence of immunoglobulin deposits and complement in the basement membrane of these patients in the early phases of the disease. CONCLUSION: The techniques used in this study have enabled us to make an earlier diagnosis of the disease, since it manifests the eye dryness immediately and the posterior biopsy can confirm the differential diagnosis. PMID- 11438879 TI - [Characteristics of scleritis in patients older than 60 years]. AB - PURPOSE: This study aimed to describe the clinical characteristics of scleritis presented de novo in patients older than 60 years of age. DESIGN: Type of scleritis, ocular manifestations, previous ocular surgery, and disease association were evaluated in patients older than 60 years with scleritis. Comparisons were made between patients with scleritis older and younger than 60 years. RESULTS: Patients with scleritis older than 60 years (61 of 195 patients, 31.3%) had more necrotizing scleritis (47.5%, p=0.0001), decrease in vision (57.4%, p=0.0001), peripheral ulcerative keratitis (29.5%, p=0.0001), previous ocular surgery (39.3%, p=0.0001), and disease association (73.8%, p=0.0001) than did patients with scleritis younger than 60 years. Rheumatoid arthritis (p=0.03) and Wegener granulomatosis (p=0.01) were the most common associated diseases. CONCLUSIONS: The detection of scleritis presented for the first time in the elderly, indicates a poor ocular prognosis because it is often associated with necrotizing scleritis, decrease in vision, and peripheral ulcerative keratitis. It has also an ominous systemic prognosis because it is often associated with potentially lethal systemic diseases. PMID- 11438880 TI - [Vitrectomy and perfluorocarbons in the management of luxated lenses]. AB - PURPOSE: To evaluate the use of vitrectomy with perfluorocarbon liquids (Perfluoro-DK-line) (PFCL) to refloat the luxated lenses of surgical etiology. Follow-up complications are considered. METHODS: The cases with vitreous posteriorly luxated lenses were reviewed between 1989 and 1999. 8 cases were found, all had similar surgery with vitrectomy and PFCL performed by the same surgeon. In two cases an anterior chamber IOL and in 6 cases a posterior chamber IOL sulcus fixated were used. RESULTS: The average age of the 8 cases with unilateral vitreous luxated lenses of no surgical etiology was 64.5 years. 3 of them were myopic with a history of spontaneous luxation with a duration of 5.9 and 10 years respectively. The other 5 cases reported a history of ocular trauma with a shorter time of luxation (between 1 week and 3 years). 7 cases (87.5%) had ocular hypertension, 6 cases (75%) showed epithelial corneal edema and one case retinal detachment preoperatively. Vitrectomy with PFCL normalized IOP and corneal edema. The complications related with decreased final VA were cystoid macular edema (1 case), macular atrophy (1 case) and retinal detachment (1 case). Only in one case a postsurgical retinal detachment was found likely to be iatrogenic. CONCLUSION: As vitrectomy with PFCL simplifies surgical technique with good results and modern sulcus fixated IOL may be used during the same surgery offering visual restoration, we believe this combined technique must be recommended always after posterior lens dislocation, to prevent complications and to preserve visual acuity. PMID- 11438881 TI - [Percutaneous topical anaesthesia applications in ocular surgery]. AB - PURPOSE/METHODS: The results of our experience with EMLA cream used as a topical anaesthetic is analysed in a series of forty patients having bilateral and simultaneous oculoplastic procedures, such as the injection of different substances (botulinum toxin, triamcinolone), the removal of superficial skin lesions, or previous to subcutaneous infiltration of local anaesthetics. Patient's reaction to pain and its degree were assessed by the use of topical placebo on the other than EMLA site and compare both subjective and <> scales. RESULTS/CONCLUSIONS: Patient's discomfort and pain were found to be milder with a statistically significant difference (p<0.0001) compared to placebo when EMLA cream was previously used over the surgical site. Therefore, in our experience, the use of EMLA cream as percutaneous anaesthetic is effective in diminishing pain associated with minor oculoplastic procedures. PMID- 11438882 TI - [Bilateral exophthalmos secondary to right carotid-cavernous sinus fistula]. AB - CASE REPORT: Male with cranioencephalic trauma who rapidly developed a marked bilateral exophthalmos. A CT was requested, the CT findings suggest a right-side carotid-cavernous sinus fistula (CCSF), which was confirmed by means of a cerebral arteriography. Following treatment with intravascular balloon pump the patient's condition improved quite spectacularly. DISCUSSION: CCSFs are anomalous communications between the cavernous sinus and the carotid artery which may be of a spontaneous nature, a result of trauma or iatrogenia and which have hemodynamic repercussions throughout the drainage area of the cavernous sinus. Sonography, CT or MR are useful whilst a cerebral angiography is required for diagnosis. CCSFs resulting from trauma normally require endovascular therapy with selective embolization techniques. PMID- 11438883 TI - [Traumatic luxation of the globe into the maxilar sinus]. AB - PURPOSE: We present a case of traumatic luxation of the ocular globe into the maxillary sinus. MATERIAL AND METHODS: Male patient, 23 years old with orbito facial trauma and traumatic luxation of his right ocular globe into the maxillary sinus. CONCLUSION: To present an unusual case of traumatic prolapse of an undamaged ocular globe into a paranasal sinus. PMID- 11438884 TI - [Instruments needed for surgery of senile cataracts (1904)]. PMID- 11438885 TI - [Allvar Gullstrand. Landskrona (Sweden), 1862 - Stockholm (Sweden), 1930]. PMID- 11438886 TI - Sequence variation and linkage disequilibrium in the human T-cell receptor beta (TCRB) locus. AB - The T-cell receptor (TCR) plays a central role in the immune system, and > 90% of human T cells present a receptor that consists of the alpha TCR subunit (TCRA) and the beta subunit (TCRB). Here we report an analysis of 63 variable genes (BV), spanning 553 kb of TCRB that yielded 279 single-nucleotide polymorphisms (SNPs). Samples were drawn from 10 individuals and represent four populations African American, Chinese, Mexican, and Northern European. We found nine variants that produce nonfunctional BV segments, removing those genes from the TCRB genomic repertoire. There was significant heterogeneity among population samples in SNP frequency (including the BV-inactivating sites), indicating the need for multiple-population samples for adequate variant discovery. In addition, we observed considerable linkage disequilibrium (LD) (r(2) > 0.1) over distances of approximately 30 kb in TCRB, and, in general, the distribution of r(2) as a function of physical distance was in close agreement with neutral coalescent simulations. LD in TCRB showed considerable spatial variation across the locus, being concentrated in "blocks" of LD; however, coalescent simulations of the locus illustrated that the heterogeneity of LD we observed in TCRB did not differ markedly from that expected from neutral processes. Finally, examination of the extended genotypes for each subject demonstrated homozygous stretches of >100 kb in the locus of several individuals. These results provide the basis for optimization of locuswide SNP typing in TCRB for studies of genotype-phenotype association. PMID- 11438888 TI - Hereditary vascular retinopathy, cerebroretinal vasculopathy, and hereditary endotheliopathy with retinopathy, nephropathy, and stroke map to a single locus on chromosome 3p21.1-p21.3. AB - We performed a genomewide search for linkage in an extended Dutch family with hereditary vascular retinopathy associated with migraine and Raynaud phenomenon. Patients with vascular retinopathy are characterized by microangiopathy of the retina, accompanied by microaneurysms and telangiectatic capillaries. The genome search, using a high throughput capillary sequencer, revealed significant evidence of linkage to chromosome 3p21.1-p21.3 (maximum pairwise LOD score 5.25, with D3S1578). Testing of two additional families that had a similar phenotype, cerebroretinal vasculopathy, and hereditary endotheliopathy with retinopathy, nephropathy, and stroke, revealed linkage to the same chromosomal region (combined maximum LOD score 6.30, with D3S1588). Haplotype analysis of all three families defined a 3-cM candidate region between D3S1578 and D3S3564. Our study shows that three autosomal dominant vasculopathy syndromes with prominent cerebroretinal manifestations map to the same 3-cM interval on 3p21, suggesting a common locus. PMID- 11438887 TI - Localization of a susceptibility gene for familial nonmedullary thyroid carcinoma to chromosome 2q21. AB - The familial form of nonmedullary thyroid carcinoma (NMTC) is a complex genetic disorder characterized by multifocal neoplasia and a higher degree of aggressiveness than its sporadic counterpart. In a large Tasmanian pedigree (Tas1) with recurrence of papillary thyroid carcinoma (PTC), the most common form of NMTC, an extensive genomewide scan revealed a common haplotype on chromosome 2q21 in seven of the eight patients with PTC. To verify the significance of the 2q21 locus, we performed linkage analysis in an independent sample set of 80 pedigrees, yielding a multipoint heterogeneity LOD score (HLOD) of 3.07 (alpha=0.42), nonparametric linkage (NPL) 3.19, (P=.001) at marker D2S2271. Stratification based on the presence of at least one case of the follicular variant of PTC, the phenotype observed in the Tas1 family, identified 17 such pedigrees, yielding a maximal HLOD score of 4.17 (alpha=0.80) and NPL=4.99 (P=.00002) at markers AFMa272zg9 and D2S2271, respectively. These results indicate the existence of a susceptibility locus for familial NMTC on chromosome 2q21. PMID- 11438889 TI - Detection of elevated serum beta-chemokine levels in seronegative Chinese individuals exposed to human immunodeficiency virus type 1. AB - The mutations in the CCR5 coding region, such as CCR5Delta32 and CCR5m303, that suppress the transmission of human immunodeficiency virus (HIV) type 1 do not exist in Chinese people. However, 9 Chinese subjects in Taiwan with histories of multiple sexual exposures to HIV remained uninfected, suggesting that certain anti-HIV factors do indeed exist. Experiments were therefore designed to investigate the immune mechanism that protects this cohort against HIV infection. Peripheral blood samples from these 9 subjects and 7 healthy people who had not been exposed to HIV were obtained for the quantitation of the levels for beta chemokines and interleukin 16 (IL-16) in serum samples or secreted by peripheral blood lymphocytes. Significantly higher serum levels for nearly all 3 beta chemokines, regulation on activation, normal T cell-expressed and secreted, macrophage inflammatory protein (MIP)-1alpha, and MIP-1beta (P<.05, P<.05, and P=.05, respectively), but not IL-16, were detected in the 9 HIV-uninfected subjects as compared with control subjects. The result suggests that among the host genes and cellular factors thus far identified, beta-chemokines are the major HIV-suppressive factors that protect Chinese people from infection with HIV. PMID- 11438890 TI - Etiology of gastroenteritis in sentinel general practices in the netherlands. AB - Data from a general practice-based, case-control study on gastroenteritis and the pathogens related to this disease were used to study the association between specific pathogens and the infected patients' ages and symptoms. For comparison, the occurrence of these pathogens in control patients, stratified by age, also is presented. In children with gastroenteritis who were <5 years of age, rotavirus (in 21% of patients) and Norwalk-like virus (NLV; in 15%) were the most common pathogens. Among patients who were 5-14 years of age, Campylobacter species (in 16% of patients) and Giardia lamblia (in 10%) were the most common pathogens. In the older patients, Campylobacter species was also the most common pathogen (8% to 15% of patients). In addition, several symptoms in case patients were associated with specific pathogens. Blood in the stool was associated with infection with Campylobacter species. In patients with fever, Salmonella species, Campylobacter species, and rotavirus were detected relatively often. Vomiting was associated with NLV and rotavirus. This is the first study in The Netherlands and one of the first studies in the world that has investigated a broad range of pathogens recovered from an unselected population of patients who had consulted general practitioners because of gastroenteritis. PMID- 11438891 TI - Impact of a hospital-based antimicrobial management program on clinical and economic outcomes. AB - Inappropriate use of antimicrobial agents results in unnecessary exposure to medication, persistent or progressive infection, emergence of resistance, and increased costs. We implemented a program to control use of restricted agents while improving care. This study compared 2 major mechanisms for improving use of antimicrobial agents: (1) recommendations made by the Antimicrobial Management Team (AMT), which included a clinical pharmacist backed up by a physician from the Division of Infectious Diseases (ID), and (2) recommendations made by ID fellows. Outcome measures included appropriateness of recommendations, cure rate, number of treatment failures, and cost of care, which were assessed for 180 patients. The AMT outperformed the ID fellows in all outcomes examined by the study (including appropriateness [87% vs. 47%; P<.001], cure rate [64% vs. 42%; P=.007], and treatment failures [15% vs. 28%; P=.03]), although the differences in economic outcomes between cases managed by the AMT and those managed by the ID fellows were not statistically significant. In an academic setting with a restricted formulary, the AMT demonstrated better antimicrobial prescribing than ID fellows. PMID- 11438892 TI - Financial and clinical impact of false-positive blood culture results. AB - During a 4-year period, 9465 specimens for blood culture were obtained from 11,911 highly febrile, otherwise healthy young children. Of these specimens, 87 (0.9%) yielded nonpathogens and were considered to be false-positive blood culture results (FPBCRs). Seventy-two of the patients who provided these specimens were treated on an outpatient basis (resulting in $10,821 in treatment charges), and 7 were admitted to the hospital (resulting in $16,200 in charges) as a result of the FPBCRs. Studies performed during follow-up included a second blood culture (21 patients), complete blood cell count (11), urine analysis and culture (5), lumbar puncture (3), and chest radiography (3). Only a small minority of patients were hospitalized or underwent invasive procedures. The charges associated with FPBCRs are very small in comparison to the initial charges of culturing specimens obtained from children considered to be at risk. Concern about the consequences of FPBCRs should not deter clinicians from performing indicated cultures for children who potentially have bacteremia. PMID- 11438893 TI - Association of tuberculin sensitivity in Dutch adults with history of travel to areas of with a high incidence of tuberculosis. AB - International travel may be a source of introduction of tuberculosis into low incidence countries. We assessed whether, in The Netherlands, sensitivity to tuberculin was associated with a history of travel to countries with a high incidence of tuberculosis. Immunocompetent adults with no history of Bacille Calmette-Guerin vaccination or sensitivity to tuberculin were skin-tested simultaneously with 1-tuberculin unit (TU) purified protein derivative (PPD) of Mycobacterium tuberculosis and 1-TU sensitin of Mycobacterium scrofulaceum. Tuberculin sensitivity was defined as a reaction to PPD of > or =10 mm that was > or =3 mm larger than the reaction to M. scrofulaceum sensitin. Tuberculin sensitivity was found in 7 (0.7%) of 1014 participants (95% confidence interval [CI], 0.3%-1.4%); it was independently associated with a cumulative history of >3 months' travel to high-incidence areas (odds ratio, 6.0; 95% CI, 1.2-31.2; P=.016) and increased in association with total duration of travel (P=.02). Travel to high-incidence areas increases the risk of tuberculin sensitivity and, consequently, of latent tuberculous infection. In countries with a low incidence of tuberculosis, cases of infection acquired during travel may account for a substantial proportion of new infections in the resident population. PMID- 11438894 TI - Mycobacterium tuberculosis complex DNA from an extinct bison dated 17,000 years before the present. AB - In order to assess the presence of tuberculosis in Pleistocene bison and the origin of tuberculosis in North America, 2 separate DNA extractions were performed by 2 separate laboratories on samples from the metacarpal of an extinct long-horned bison that was radiocarbon dated at 17,870+/-230 years before present and that had pathological changes suggestive of tuberculosis. Polymerase chain reaction amplification isolated fragments of tuberculosis DNA, which were sequenced, and on which spoligotyping was also performed to help determine its relationship to the various members of the Mycobacterium tuberculosis complex. Extensive precautions against contamination with modern M. tuberculosis complex DNA were employed, including analysis of paleontologic and modern specimens in 2 geographically separate laboratories. PMID- 11438895 TI - Risks factors and prevention of Q fever endocarditis. AB - Coxiella burnetii causes acute and chronic Q fever. To evaluate the risk factors of development of chronic endocarditis following Q fever and to assess the best preventive therapy, a retrospective study of patients diagnosed as having Q fever during 1985-2000 was conducted. Twelve patients with acute Q fever who developed endocarditis and 102 patients with Q fever endocarditis were included in the study. When compared to 200 control patients with acute Q fever, preexisting valvular disease (P<10(-7)), especially a prosthetic valve (P=.01), were encountered more often among patients with endocarditis. Among patients with valvular defects, we estimate the risk of developing endocarditis to be 39%. A combination of doxycycline plus hydroxychloroquine was better at preventing the development of endocarditis than doxycycline alone (P=.009). Our results should encourage physicians to detect valvular lesions in patients with acute Q fever and to search for acute Q fever in patients with a valvulopathy and unexplained fever. A proper treatment for such patients and a scheduled follow-up should reduce the risk of developing endocarditis. PMID- 11438896 TI - Outbreak of sternal surgical site infections due to Pseudomonas aeruginosa traced to a scrub nurse with onychomycosis. AB - From 19 February 1999 through 31 October 1999, 16 (8.6%) of 185 patients who underwent median sternotomy developed infections with Pseudomonas aeruginosa. Seven patients had mediastinitis, 5 had deep sternal wound infection, 2 had superficial sternal wound infection, 1 had prosthetic valve endocarditis, and 1 had sepsis. Pulsed-field gel electrophoresis confirmed that all 13 isolates that were available for typing were the same strain. Cultures of hand specimens identified 1 nurse from whom the same strain of P. aeruginosa was repeatedly isolated; the nurse had been in contact with all 16 infected patients. Investigation revealed that the nurse had severe onycholysis and onychomycosis of the right thumbnail. Cultures of samples of this nail's subungual region and of multiple cosmetic products from the nurse's home yielded the identical P. aeruginosa strain. This outbreak of surgical site infections due to P. aeruginosa was caused by wound contamination from the thumbnail of this nurse, despite her appropriate use of latex surgical gloves. PMID- 11438897 TI - Antimicrobial resistance prevalence rates in hospital antibiograms reflect prevalence rates among pathogens associated with hospital-acquired infections. AB - To determine whether routine antibiograms (summaries reporting resistance of all tested isolates) reflect resistance rates among pathogens associated with hospital-acquired infections, we compared data collected from 2 different surveillance components in the same 166 intensive care units (ICUs). ICUs reported data during the same months to both the infection-based surveillance and the laboratory-based surveillance. Paired comparisons of the percentage of isolates resistant were made between systems within each ICU. No significant differences existed (P>.05) between the percentage of isolates resistant from the infection-based system and laboratory-based system for all antimicrobial resistant organisms studied, except methicillin resistance in Staphylococcus species. The mean difference in percentage resistance was higher from the infection-based system than the laboratory-based system for S. aureus (mean difference, +8%, P<.001) and coagulase-negative staphylococci (mean difference, +9%, P<.001). Overall, hospital antibiograms reflected susceptibility patterns among isolates associated with hospital-acquired infections. Hospital antibiograms may underestimate the relative frequency of methicillin resistance among Staphylococcus species when associated with hospital-acquired infections. PMID- 11438898 TI - Mycobacterium haemophilum in immunocompromised patients. AB - Mycobacterium haemophilum, a recently described pathogen, can cause an array of symptoms in immunocompromised patients. To date, 90 patients with this infection have been described worldwide. We report our institution's experience with 23 patients who were treated from 1990 through 2000. Fourteen patients had undergone bone marrow transplantation, 5 were infected with human immunodeficiency virus, 3 had hematologic malignancies, and 1 had no known underlying immunosuppression. Clinical syndromes on presentation included skin lesions alone in 13 patients, arthritis or osteomyelitis in 4 patients, and lung disease in 6 patients. Although patients with skin or joint involvement had favorable outcomes, 5 of 7 patients with lung infection died. Prolonged courses of multidrug therapy are required for treatment. A diagnosis of M. haemophilum infection must be considered for any immunocompromised patient for whom acid-fast bacilli are identified in a cutaneous, synovial fluid or respiratory sample or for whom granulomas are identified in any pathological specimen. PMID- 11438899 TI - Risk factors for anaerobic bloodstream infections in bone marrow transplant recipients. AB - The incidence of anaerobic bloodstream infections (BSI) in patients who underwent bone marrow transplantation (BMT) recently increased at our institution. A retrospective case-control study of patients undergoing BMT from January 1995 through December 1998 was performed to determine the microbiological characteristics, epidemiology, and outcome of anaerobic BSI and to identify independent risk factors for infection. Anaerobic BSI occurred in 23 patients, for a rate of 4 BSIs per 100 BMT procedures, and it accounted for 17% of all BSIs that occurred during the study period. Infection occurred at a mean (+/- standard deviation) of 7+/-4 days after BMT and 7+/-5 days after the onset of neutropenia. Fusobacterium nucleatum was the most frequently isolated pathogen (in 17 patients), followed by Leptotrichia buccalis (in 4), Clostridium septicum (in 1), and Clostridium tertium (in 1). Two case patients (9%) died. Severity of mucositis was an independent predictor of anaerobic BSI (odds ratio, 4.4; P=.01). Controlling mucositis is critical for the prevention of anaerobic BSI in this patient population. PMID- 11438900 TI - Effect of cessation of highly active antiretroviral therapy during a discordant response: implications for scheduled therapeutic interruptions. AB - Although treatment with combination antiretroviral therapy leads to a reduction in the level of plasma viremia and an improvement in CD4 T cell count for most patients, for a minority of patients, an improvement in CD4 T cell count occurs despite the failure of treatment to suppress viral replication. Recent reports suggest that these discordant improvements in CD4 T cell count may last for months to years and are associated with improved clinical outcomes. In a retrospective observational study, we evaluated the effect of therapy cessation on 8 patients with discordant immunologic responses to therapy and found that improved CD4 T cell responses are dependent upon ongoing drug pressure. If antiretroviral agents that are likely to resuppress the virus are not available, we suggest that patients continue the therapy associated with immunologic improvement to maximize the clinical benefit of the discordant response. PMID- 11438901 TI - Clinical features of clostridial bacteremia: a review from a rural area. AB - Blood samples, which were obtained from patients who lived in a rural area with approximately 500 acute-care hospital beds, were cultured from 1990 through 1997. We retrospectively reviewed the blood cultures that yielded Clostridium species (74 [0.12%] of 63,296 cultures). These were obtained from 46 different hospitalized patients (incidents per hospital, 0.03%). The source of the Clostridium species was a gastrointestinal site in 24 patients (52.2%). The most frequently identified Clostridium species was Clostridium perfringens (in 10 [21.7%] of patients), followed by Clostridium septicum (in 9 [19.6%]). Thirty-one patients (67.4%) were aged > or =65 years, 13 patients (28.3%) had diabetes mellitus, and underlying malignancy was present in 22 patients (47.8%). The mortality rate of patients whose condition had been managed surgically was 33%; for those patients whose conditions required medical management, the mortality rate was 58%. Clostridium bacteremia in these patients usually had a gastrointestinal source, it often occurred in patients with serious underlying medical conditions, and it rarely was the result of traumatic farm accidents. PMID- 11438902 TI - Pathogenesis of maternal-fetal syphilis revisited. AB - Although congenital syphilis has been recognized for several centuries and an efficient treatment with penicillin became available more than a half-century ago, the disease is still with us. Inability to culture in vitro the causative agent, Treponema pallidum, and the lack of an adequate animal model have prevented exploration of the various immunopathological events affecting the natural course of congenital infection. The purpose of this review is to analyze the disease in the context of recent knowledge acquired from human and experimental animals, particularly from the guinea pig model of congenital and neonatal syphilis, and to describe how the infection interacts with the maternal fetal unit and how it is further modulated by the conceptus' ontogenic development. We also attempt to elucidate several old immunologic concepts and misconceptions that have remained unchallenged for too long. PMID- 11438903 TI - Dissemination of antibiotic-resistant bacteria across geographic borders. AB - The development of antibiotic-resistant (AR) bacteria in any country is of global importance. After their initial selection and local dissemination, AR bacteria can be transferred across international borders by human travelers, animal and insect vectors, agricultural products, and surface water. The sources and routes of importation of strains of AR bacteria are most often unknown or undetected, because many bacteria carrying resistance genes do not cause disease, and routine surveillance often does not detect them. Control of international dissemination of AR bacteria depends on methods to reduce selection pressure for the development of such bacteria and improved surveillance to detect their subsequent spread. PMID- 11438904 TI - Congenital lymphocytic choriomeningitis virus infection: decade of rediscovery. AB - Lymphocytic choriomeningitis virus (LCMV) is an underdiagnosed fetal teratogen. This diagnosis should be considered for infants and children with unexplained hydrocephalus, micro- or macrocephaly, intracranial calcifications, chorioretinitis, and nonimmune hydrops. The immunofluorescent antibody test is the only reasonable, commercially available, screening diagnostic tool. The differential diagnosis of congenital LCMV infection includes toxoplasmosis, rubella, cytomegalovirus, herpes simplex virus, enteroviruses, human parvovirus B19 [corrected], and syphilis. The infection has also been misdiagnosed as various neurologic, ophthalmologic, and chromosomal syndromes. Further research, to determine the prevalence of this infection in human and rodent populations, and prospective studies, to delineate the clinical spectrum of congenital infection, are needed. The public and members of the medical profession should be made aware of the hazard that wild, pet, and laboratory rodents pose to pregnant women. PMID- 11438906 TI - Food irradiation: a public health challenge for the 21st century. AB - Although the United States food supply is generally considered one of the safest in the world, foodborne illnesses remain a concern. Each year, millions of Americans become ill and as many as 5000 die from foodborne infections. The United States Department of Agriculture estimates that medical treatment and productivity losses associated with foodborne illnesses cost as much as $37 billion annually. Irradiation, which involves exposing food briefly to radiant energy, can reduce or eliminate microorganisms that contaminate food or cause spoilage. So far, only limited quantities of irradiated foods--spices, herbs, dry vegetable seasonings, and some fresh fruits, vegetables, and poultry--have been available in the United States. Major purchasers are health care and food service establishments. The World Health Organization reviewed 500 studies and concluded that food irradiation poses no toxicological, microbiological, or nutritional problems. In more than 40 years, there have been no accidents in North America involving transport of the types of radioactive isotopes used for irradiation. PMID- 11438907 TI - Food irradiation: unresolved issues. AB - Food-associated diarrheal disease is a major problem in the United States. One approach offered as virtually a panacea is food irradiation. Irradiating our food would have some unequivocal benefits, but there are some important unresolved issues that must be addressed before we commit our society to a technology that could be harmful. PMID- 11438908 TI - Malaria chemoprophylaxis in the age of drug resistance. II. Drugs that may be available in the future. AB - All current regimens of malaria chemoprophylaxis have serious drawbacks as a result of either suboptimal efficacy, difficulty with medication compliance, or adverse events. Two 8-aminoquinolines may be approaching registration, with primaquine having completed its prophylactic field testing and tafenoquine having begun advanced field testing at the end of 2000. Primaquine has long been used for management of relapses of malaria, but in the past decade, it has been reexamined for use in malaria prevention in order to stop infection in the liver. In field trials performed in Indonesia and Colombia, the efficacy of primaquine for malaria prevention was approximately 90%, compared with that of placebo. Because of its short half-life, primaquine requires daily administration. For adults, the prevention regimen is 30 mg base daily (0.5 mg base/kg/day), and it can probably be discontinued soon after departure from an area where malaria is endemic. To kill parasites that already exist in the liver, terminal prophylaxis is given after exposure to relapses of malaria infection; for adults, such prophylaxis usually consists of 15 mg base (0.3 mg base/kg/day) given daily for 2 weeks. Primaquine-induced gastrointestinal disturbances can be minimized if the drug is taken with food. Neither primaquine nor tafenoquine should be given to persons with glucose-6-phosphate dehydrogenase deficiency, to avoid the development of potentially severe drug-induced hemolysis. Tafenoquine is an analogue of primaquine that is more potent than the parent drug. Field trials in Kenya, Ghana, Gabon, and Southeast Asia have demonstrated an efficacy rate of approximately 90% for tafenoquine. Its long half-life allows for infrequent dosing (currently tested at 200 mg base/week), and its effect on parasites at the liver stage may allow for drug discontinuation at the time of departure from the area of endemicity. PMID- 11438909 TI - Value of patient self-report and plasma human immunodeficiency virus protease inhibitor level as markers of adherence to antiretroviral therapy: relationship to virologic response. AB - Three methods of adherence to antiretroviral therapy were evaluated for 149 patients infected with human immunodeficiency virus (HIV): plasma level of protease inhibitors (PIs), patient self-report, and routine biological parameters associated with the use of some antiretroviral drugs. Adherence to therapy was estimated from a score calculated from answers to a self-administered questionnaire and on the basis of measurement of relevant plasma and blood levels. Of the 149 patients, 112 had a virologic response, and 122 had adequate trough PI levels. Plasma PI levels and virologic outcome were significantly correlated (P<.0001). The adherence score was significantly correlated with virologic response (P<.001). Macrocytosis was significantly associated with virologic response in the patients treated with zidovudine or stavudine (P=.006). PI level was the higher significant predictor of virologic response (P=.0003). Self-reported adherence (P=.01) and macrocytosis (P=.05) were also independently associated with antiretroviral efficacy. PMID- 11438910 TI - Prophylaxis with trimethoprim-sulfamethoxazole for human immunodeficiency virus infected patients: impact on risk for infectious diseases. AB - Trimethoprim-sulfamethoxazole (TMP-SMZ) is widely prescribed as prophylaxis for Pneumocystis carinii pneumonia (PCP) in human immunodeficiency virus (HIV) infected persons. Its efficacy against other infections has not been thoroughly evaluated. To compare the risk for infectious diseases for persons who were prescribed TMP-SMZ with that for patients who were not prescribed TMP-SMZ, we examined data collected from the medical records of HIV-infected patients (January 1990 through September 1999) who were enrolled in the Adult and Adolescent Spectrum of HIV Disease Project. During intervals when patients had CD4(+) T lymphocyte counts of <200 cells/microL (19,081 persons; 22,801 person years), prescription of TMP-SMZ was associated with significant protection from toxoplasmosis, salmonellosis, infection with Haemophilus species, invasive or any staphylococcal infection, and PCP, but not from Shigella, pneumococcal or nonpneumococcal Streptococcus, Klebsiella, or Pseudomonas species. We demonstrate that prescription of TMP-SMZ for PCP prophylaxis in persons with HIV infection is associated with significantly decreased risk for several infectious diseases. These findings may be of interest to HIV prevention programs in resource-poor countries. PMID- 11438911 TI - Sexually transmitted Q fever. AB - We report the sexual transmission of Coxiella burnetii from a man with occupationally acquired Q fever to his wife. Fifteen days after coitus, his wife also developed serologically proven acute Q fever. C. burnetii DNA sequences were detected by polymerase chain reaction (PCR) performed on semen samples obtained from the husband at 4 and 15 months after the onset of acute Q fever, but PCR results were variable at 23 months, indicating the presence of few organisms. PMID- 11438912 TI - Fatal submassive hepatic necrosis associated with tyrosine-methionine-aspartate aspartate-motif mutation of hepatitis B virus after long-term lamivudine therapy. AB - We present a case of infection with lamivudine-resistant mutant hepatitis B virus (HBV) that fatally exacerbated hepatitis following the emergence of HBV with mutations in the tyrosine-methionine-aspartate-aspartate (YMDD) motif in an immunocompetent patient who was receiving long-term lamivudine therapy. Restriction fragment length polymorphism analysis showed that the YMDD-motif mutant was the predominant form of circulating HBV at the time of the fatal exacerbation, and a necropsy specimen of the liver revealed submassive hepatic necrosis without steatosis. PMID- 11438913 TI - Lack of sensitivity of the latex agglutination test to detect bacterial antigen in the cerebrospinal fluid of patients with culture-negative meningitis. AB - For culture-negative meningitis, use of the latex agglutination test for detection of bacterial antigen in the CSF has a sensitivity of only 7%. Routine use of the latex agglutination test may not contribute to the management of suspected acute bacterial meningitis, since patients are treated on the basis of their clinical presentations and CSF findings. PMID- 11438914 TI - Efficacy and safety of clarithromycin as treatment for Mediterranean spotted fever in children: a randomized controlled trial. AB - Fifty-one children with Mediterranean spotted fever (MSF) were randomized to receive either clarithromycin, 15 mg/kg/day orally in 2 divided doses, or chloramphenicol, 50 mg/kg/day orally in 4 divided doses, for 7 days. Mean time to defervescence was 36.7 h in the clarithromycin group and 47.1 h in the chloramphenicol group (P=.047). Clarithromycin could be an acceptable therapeutic alternative to chloramphenicol and to tetracyclines for children aged <8 years with MSF. PMID- 11438915 TI - False-positive results of PCR testing for Lyme disease. PMID- 11438916 TI - Disorders of iron metabolism associated with protease inhibitor therapy. PMID- 11438917 TI - Alendronate treatment for osteoporosis in patients infected with human immunodeficiency virus. PMID- 11438918 TI - Bacteremia and meningitis caused by a macrolide-sensitive strain of Streptococcus pneumoniae during treatment with azithromycin. PMID- 11438919 TI - Pseudomonas pneumonia in smokers. PMID- 11438920 TI - Focal inhibitory interneuron loss and principal cell hyperexcitability in the rat hippocampus after microinjection of a neurotoxic conjugate of saporin and a peptidase-resistant analog of Substance P. AB - Episodes of prolonged seizures or head trauma produce chronic hippocampal network hyperexcitability hypothesized to result primarily from inhibitory interneuron loss or dysfunction. The possibly causal role of inhibitory neuron failure in the development of epileptiform pathophysiology remains unclear because global neurologic injuries produce such a multitude of effects. The recent finding that Substance P receptors (SPRs) are expressed exclusively in the rat hippocampus by inhibitory interneurons provided the rationale for attempting to ablate interneurons selectively by using neurotoxic conjugates of SPR ligands and the ribosome inactivating protein saporin that specifically target Substance P receptor-expressing cells. Whereas intrahippocampal microinjection of a conjugate of native SP and saporin produced significant nonspecific damage at concentrations needed to produce even limited selective loss of SPR-positive cells, a conjugate of saporin and the more potent and peptidase-resistant SP analog [Sar(9), Met(O(2))(11)] Substance P (SSP-saporin) caused negligible nonspecific damage at the injection site, and a virtually complete loss of SPR like immunoreactivity (LI) up to 1 mm from the injection site. Within the SPR depletion zone, immunoreactivities for most GABA-, parvalbumin-, somatostatin-, and cholecystokinin-immunoreactive cells and fibers were eliminated. The few interneurons detectable within the affected zone were devoid of SPR-LI. The apparent loss of interneurons was selective in that calbindin- and glutamate receptor subunit 2 (GluR2) -positive principal cells survived within the affected zone, as did myelinated fibers and the extrinsic calretinin- and tyrosine hydroxylase--immunoreactive terminals of subcortical afferents. An apparent lack of reactive synaptic reorganization in response to interneuron loss was indicated by zinc transporter-3 (ZnT3)-- and beta-synuclein--LI, as well as by Timm staining, all of which revealed relatively normal patterns of excitatory terminal distribution. Control injections produced minor damage at the injection site, but no apparent specific loss of SPR-LI. One to 12 weeks after injection of SSP saporin, extracellular electrophysiological field responses recorded in the CA1 pyramidal and dentate granule cell layers in response to afferent stimulation were blindly evaluated simultaneously in two sites 1-2 mm apart along the longitudinal hippocampal axis. SSP-saporin-treated rats exhibited relatively normal responses in some sites, whereas disinhibition and hyperexcitability indistinguishable from the pathophysiology produced by experimental status epilepticus were simultaneously recorded at adjacent sites. Anatomic analysis of the recording sites in each animal revealed that epileptiform pathophysiology was consistently observed only within areas of SPR ablation, whereas relatively normal evoked responses were recorded from immediately adjacent and relatively unaffected regions. These data establish the efficacy of [Sar(9), Met(O(2))(11)] Substance P-saporin for producing a selective and spatially extensive ablation of hippocampal inhibitory interneurons in vivo and a highly focal disinhibition that was restricted to the site of interneuron loss. These results also demonstrate that the "epileptic" pathophysiology produced by experimental status epilepticus or head trauma can be replicated by focal interneuron loss per se, without involving principal cell loss and other interpretive confounds inherent in the use of global neurologic injury models. PMID- 11438921 TI - Nucleus isthmi, pars semilunaris as a key component of the tectofugal visual system in pigeons. AB - The avian isthmic nuclei are constituted by a group of structures reciprocally connected with the tectum opticum and considered to play a role in the modulation of intratectal processes. Although the two larger isthmic nuclei, the n. isthmi pars parvocellularis (Ipc) and the n. isthmi pars magnocellularis (Imc), have been studied in detail previously, the third and smallest of this group, the n. isthmi pars semilunaris (SLu), has been largely neglected. The present study demonstrates this isthmic component to be characterized by a unique connectivity and immunohistochemical pattern: 1) SLu receives tectal afferents and projects back onto the outer retinorecipient tectal layers; 2) it projects bilaterally onto the nucleus rotundus and thus modulates the ascending tectofugal system; 3) in addition, previous studies have demonstrated SLu projections onto the lateral spiriform nucleus (SpL), which mediates basal ganglia output onto the tectum. In that SpL projects onto the deep layers of the tectum, SLu indirectly modulates descending tectal output patterns. Taken together, the role of SLu goes far beyond a local modulation of intratectal processes. Instead, this isthmic structure is likely to play a key role in the topographically organized modulation of the ascending and, at least indirectly, also the descending projections of the optic tectum. PMID- 11438922 TI - Expression patterns of Notch1, Notch2, and Notch3 suggest multiple functional roles for the Notch-DSL signaling system during brain development. AB - The Notch-DSL signaling system consists of multiple receptors and ligands, and plays many roles in development. The function of Notch receptors and ligands in mammalian brain, however, is poorly understood. In the current study, we examined the expression patterns for three receptors of this system, Notch1, 2, and 3, in late embryonic and postnatal rat brain by in situ hybridization. The three receptors have overlapping but different patterns of expression. Messenger RNA for all three proteins is found in postnatal central nervous system (CNS) germinal zones and, in early postnatal life, within numerous cells throughout the CNS. Within zones of cellular proliferation of the postnatal brain, Notch1 mRNA is found in both the subventricular and the ventricular germinal zones, whereas Notch2 and Notch3 mRNAs are more highly localized to the ventricular zones. Both Notch1 and Notch3 mRNAs are expressed along the inner aspect of the dentate gyrus, a site of adult neurogenesis. Notch2 mRNA is expressed in the external granule cell layer of the developing cerebellum. In several brain areas, Notch1 and Notch2 mRNAs are relatively concentrated in white matter, whereas Notch3 mRNA is not. Neurosphere cultures (which contain CNS stem cells), purified astrocyte cultures, and striatal neuron-enriched cultures express Notch1 mRNA. However, in these latter cultures, Notch1 mRNA is produced by nestin-containing cells, rather than by postmitotic neurons. Taken together, these results support multiple roles for Notch1, 2, and 3 receptor activation during CNS development, particularly during gliogenesis. PMID- 11438923 TI - Microglia and the early phase of immune surveillance in the axotomized facial motor nucleus: impaired microglial activation and lymphocyte recruitment but no effect on neuronal survival or axonal regeneration in macrophage-colony stimulating factor-deficient mice. AB - Activation of microglia is among the first cellular changes in the injured CNS. However, little is known about their specific contribution to secondary damage or repair processes in neighboring neurons and nonneuronal cells or to the immune surveillance of the damaged tissue. Animal models with defective microglial response such as osteopetrosis provide an approach to explore these effects. Osteopetrosis (op) is an autosomal recessive mutation with a complete deficiency of the macrophage-colony stimulating factor (MCSF; CSF-1), an important mitogen for brain microglia. In the current study we examined the effects of this MCSF deficiency on the microglial reaction and the overall cellular response to nerve injury in the mouse axotomized facial motor nucleus. In the brain, MCSF receptor immunoreactivity was found only on microglia and was strongly up-regulated following injury. MCSF deficiency led to a failure of microglia to show a normal increase in early activation markers (thrombospondin, MCSF receptor, alpha M beta 2- and alpha 5 beta 1-integrins), to spread on the surface of axotomized motoneurons, and to proliferate after injury. Early recruitment of CD3(+) T lymphocytes to the facial nucleus 24 hours after injury was reduced by 60%. In contrast, the neuronal and astrocyte response was not affected. There was a normal increase in the neuropeptides calcitonin gene-related peptide and galanin, neuronal c-JUN, and NADPH-diaphorase and a decrease in choline acetyltransferase and acetylcholinesterase. Astrocyte glial fibrillary acidic protein immunoreactivity also showed a normal increase. There was a normal influx of macrophages and granulocytes into the injured facial nerve. Synaptic stripping, neuronal survival, and speed of axonal regeneration were also not affected. The current results show a strong, selective effect of MCSF on the early activation of microglia and, indirectly, on lymphocyte recruitment. This early phase of microglial activation appears not to be involved in the process of repair following peripheral nerve injury. However, it is important in the initiation of inflammatory changes in the brain and in the interaction with the immune system. PMID- 11438924 TI - Major difference in the expression of delta- and mu-opioid receptors between turtle and rat brain. AB - The reptilian turtle brain has a remarkably higher endurance for anoxia than mammalian brains. Since the response to O(2) deprivation is dependent in a major way on the expression and regulation of membrane proteins, differences in such proteins may play a role in the species-related differences in hypoxic responses. Because opioid system is involved in the regulation of hypoxic responses, we asked whether there are differences between rat and turtle brains in terms of opioid receptor expression. In this work, we compared the expression and distribution of delta-and mu-opioid receptors in the turtle and rat brains. Our results show that (1) the dissociation constant (K(d)) for delta-receptor binding was approximately four times lower and B(max) was more than double in the turtle brain homogenates than in rat ones; (2) the delta-receptor binding density was heterogeneously distributed in the turtle brain, with a higher density in the rostral regions than in the brainstem and spinal cord, and was generally much higher than in rat brains from the cortex to spinal cord; (3) the delta-opioid receptors in the rat brains were mostly located in the cortex, caudate putamen, and amygdala with an extremely low density in most subcortical (e.g., hippocampus and thalamus) and almost all brainstem regions; and (4) in sharp contrast to delta-opioid receptors, mu-opioid receptor density was much lower in all turtle brain regions compared with the rat ones. Our results demonstrate that the turtle brain is actually an organ of delta-opioid receptors, whereas the rat brain has predominantly mu-opioid receptors. Because we have recently found that delta opioid receptors protect neurons against glutamate and hypoxic stress, we speculate that the unique pattern of delta-receptor receptor expression and distribution plays a critical role in the tolerance of turtle brain to stressful situations characterized by glutamate excitotoxicity. PMID- 11438925 TI - Palladin is expressed preferentially in excitatory terminals in the rat central nervous system. AB - Palladin is a recently described intracellular protein associated with the actin cytoskeleton and cell adhesion in fibroblasts. In Western and Northern blot analyses, palladin expression is ubiquitous in embryonic mice, but it is down regulated dramatically in most adult tissues. Significant amounts of palladin persist in the brain of adult rodents, as assessed by Western blot analysis. With this work, we extend preliminary observations and determine the overall distribution and subcellular location of palladin throughout the rat brain. In sagittal and coronal sections of the central nervous system, immunostain for palladin is present throughout the brain and spinal cord, but not uniformly. The densest regions of immunostain include the olfactory bulb, cerebral and cerebellar cortex, hippocampus, amygdala, superior colliculus, and superficial laminae of the spinal dorsal horn. Because immunostain characteristically is punctate, we performed double staining for palladin and the presynaptic marker synaptophysin. Confocal microscopy showed that palladin-immunopositive puncta are also immunopositive for synaptophysin; the proportion of synaptophysin immunopositive puncta that also stained for palladin ranged from 100% of mossy fiber terminals in field CA3 of the hippocampus and in the cerebellar cortex to 60--70% of terminals in the cerebral cortex, striatum, and spinal dorsal horn. The presence of palladin in synaptic terminals was confirmed by electron microscopy. Because immunostained terminals commonly establish asymmetric synapses, the selectivity of palladin expression in synaptic terminals was tested by double staining for palladin and gamma-aminobutyric acid. The modest level of colocalization in this material at both the light microscopic and electron microscopic levels suggests a selectivity of palladin for terminals that release excitatory neurotransmitters. As concomitant work in cell cultures has shown that palladin participates in axonal development and migration, the present results suggest that palladin persists at excitatory synapses of the adult nervous system. PMID- 11438926 TI - Vanilloid receptor VR1 is both presynaptic and postsynaptic in the superficial laminae of the rat dorsal horn. AB - Terminals in the rat spinal cord that express the vanilloid receptor VR1 are from small and medium dorsal root ganglion (DRG) neurons and appear prominent in lamina I and inner lamina II. Because primary afferents from these neurons can be myelinated or unmyelinated and their terminals in these laminae can be of various morphological and functional types, we undertook this study to identify the type(s) of VR1-positive afferent fibers and terminals. In the DRG, many small and medium-sized neurons are immunopositive. Under electron microscopy, dorsal root afferents that are immunopositive for VR1 are predominantly unmyelinated. Large numbers of VR1-positive terminals in lamina I are of the nonglomerular type and may contain dense core vesicles. VR1 immunoreactivity in terminals in lamina I is in good agreement with data on noxious, heat-sensitive neurons in the dorsal horn. Two types of glomerular afferent terminals in lamina II also are immunopositive for VR1. In both laminae, most VR1-positive terminals are distinct from substance P-positive terminals. However, the immunoreactivity in lamina II also is prominent in dendrites that are contacted by primary afferent endings. Because we also observed patchy immunostaining in cell bodies in lamina II, this unexpected result may reflect synthesis of VR1 by neurons in this lamina. However, because dorsal rhizotomy abolishes VR1 staining in both laminae I and II, it is suggested that the expression and intracellular dynamics of VR1 in lamina II neurons are controlled by presynaptic input. PMID- 11438927 TI - Heparan sulfate proteoglycan expression in the optic chiasm of mouse embryos. AB - Previous studies have demonstrated that heparan sulfate (HS) proteoglycans (PGs) regulate neurite outgrowth through binding to a variety of cell surface molecules, extracellular matrix proteins, and growth factors. The present study investigated the possible involvement of HS-PGs in retinal axon growth by examining its expression in the retinofugal pathway of mouse embryos by using a monoclonal antibody against the HS epitope. Immunoreactive HS was first detected in all regions of the retina at embryonic day (E) 11. The staining was gradually lost in the central regions and restricted to the retinal periphery at later developmental stages (E12--E16). Prominent staining for HS was consistently found in the retinal fiber layer and at the optic disk, indicating a possible supportive role of HS-PGs in axon growth in the retina. At the ventral diencephalon, immunostaining for HS was first detected at E12, before arrival of any retinal axons. The staining matched closely the neurons that are immunopositive for the stage-specific embryonic antigen 1 (SSEA-1). At E13 to E16, when axons are actively exploring their paths across the chiasm, immunoreactivity for HS was particularly intense at the midline. This characteristic expression pattern suggests a role for HS-PGs in defining the path of early axons in the chiasm and in regulating development of axon divergence at the midline. Furthermore, HS immunoreactivity is substantially reduced at regions flanking both sides of the midline, which coincides spatially to the position of actin-rich growth cones from subpial surface to the deep regions of the optic axon layer at the chiasm. Moreover, at the threshold of the optic tract, immunoreactive HS was localized to deep parts of the fiber layer. These findings indicate that changes in age-related fiber order in the optic chiasm and optic tract of mouse embryos are possibly regulated by a spatially restricted expression of HS-PGs. PMID- 11438928 TI - Development and reorganization of corticospinal projections in EphA4 deficient mice. AB - The Eph family of receptor tyrosine kinases and their ligands, the ephrins, are important regulators of axon guidance and cell migration in the developing nervous system. Inactivation of the EphA4 gene results in axon guidance defects of the corticospinal tract, a major descending motor pathway that originates in the cortex and terminates at all levels of the spinal cord. In this investigation, we report that although the initial development of the corticospinal projection is normal through the cortex, internal capsule, cerebral peduncle, and medulla in the brain of EphA4 deficient animals, corticospinal axons exhibit gross abnormalities when they enter the gray matter of the spinal cord. Notably, many corticospinal axons fail to remain confined to one side of the spinal cord during development and instead, aberrantly project across the midline, terminating ipsilateral to their cells of origin. Given the possible repulsive interactions between EphA4 and one of its ligands, ephrinB3, this defect could be consistent with a loss of responsiveness by corticospinal axons to ephrinB3 that is expressed at the spinal cord midline. Furthermore, we show that EphA4 deficient animals exhibit ventral displacement of the mature corticospinal termination pattern, suggesting that developing corticospinal axons, which may also express ephrinB3, fail to be repelled from areas of high EphA4 expression in the intermediate zone of the normal spinal cord. Taken together, these results suggest that the dual expression of EphA4 on corticospinal axons and also within the surrounding gray matter is very important for the correct development and termination of the corticospinal projection within the spinal cord. PMID- 11438929 TI - Embryonic central nervous system angiogenesis does not involve blood-borne endothelial progenitors. AB - We asked, whether, in the blood of avian embryos, endothelial precursor cells circulate that actually contribute to the growing vascular system in and around the central nervous system (CNS). We compared the morphology and distribution of QH1-positive cells after transplantation of quail paraxial mesoderm, after blood transfusion, in quail-chick parabiosis, or after quail bone-marrow transplantation. After head mesoderm transplantation from quail to chick, we observed sprouting endothelial cells (ECs), capillary tube formation, and chimeric endothelial lining of large arteries in the host brain. These QH1 positive quail cells showed EC morphologies that demonstrated three different aspects of CNS angiogenesis: invasion by means of filopodia, clonal proliferation and tube formation, and integration into preexisting EC layers. After blood transfusion or in chick-quail parabiosis, blood-borne QH1+ cells were found in the lumen of but not integrated into the wall of the host vascular system. Neither were QH1+ cells observed in the capillary walls of parabiotic chick chorioallantoic membranes. In both cases, the quail cells showed typical macrophage morphology. In chicks that had received quail bone marrow transplants onto their chorioallantoic membranes, QH1+ cells with macrophage, but not EC shape were occasionally seen near the inoculation site. We conclude that (1) blood-borne cells do not become ECs or directly contribute to angiogenesis inside, or in vascular plexuses around the CNS during embryonic development; (2) blood-borne cells do not contribute to the intraneural macrophage population of the embryonic CNS. PMID- 11438930 TI - Presynaptic kainate receptors in primary afferents to the superficial laminae of the rat spinal cord. AB - Subunits of glutamate receptors participate in the regulation of sensory transmission at primary afferent synapses in the superficial laminae of dorsal horn (DH). We report here on the distribution of kainate receptors (GluR5/6/7) in these laminae by using light microscope (LM) and electron microscope (EM) immunocytochemistry. Standard (4%) paraformaldehyde fixation resulted in immunostaining for GluR5/6/7 in perikarya and fine processes in lamina II, especially its inner part (IIi). Preembedding EM revealed immunostaining of dendrites, perikarya, and occasional terminals, presumed to be from primary afferent fibers, at the center of glomerular arrangements. In rats perfused with 0.5% paraformaldehyde, LM showed a more punctate staining, mainly in the ventral part of lamina IIi and lamina III, than in material fixed with 4% paraformaldehyde. Approximately two-thirds of GluR5/6/7 puncta were also immunostained with synaptophysin, suggesting that in material fixed with 0.5% paraformaldehyde, a large fraction of these are synaptic terminals. Double immunostained puncta disappear 4 days after dorsal rhizotomy, suggesting that most of GluR5/6/7-immunopositive terminals are from primary afferent fibers. EM material fixed with 0.5% paraformaldehyde confirmed the expression of GluR5/6/7 in numerous synaptic endings with morphology of primary afferents. To determine the type of primary afferent terminals that express GluR5/6/7, two neuroanatomic tracers were injected in the sciatic nerves. The lectin from Bandeiraea simplicifolia (IB4) is selectively taken up by unmyelinated primary afferent fibers that terminate in the outer part of lamina II (IIo) and dorsal part of lamina IIi, whereas the B subunit of the cholera toxin (CTB) is selectively taken up by a broader class of primary afferents which, in superficial DH, terminate mainly in laminae I, ventral part of IIi, and III. Approximately 20% of GluR5/6/7 immunoreactive puncta colocalized with IB4, whereas approximately 40% of GluR5/6/7-immunoreactive puncta colocalized with CTB. The present study shows that (1) GluR5/6/7 does not have a clear and consistent spatial relation with postsynaptic sites, (2) a large number of primary afferents express GluR5/6/7, and (3) these are not limited to one functional class. Thus, modulation by glutamate of primary afferent terminals by means of kainate receptors in the superficial laminae of DH may predominantly involve presynaptic mechanisms. PMID- 11438931 TI - Projections of the pontine nuclei to the cochlear nucleus in rats. AB - In the cochlear nucleus, there is a magnocellular core of neurons whose axons form the ascending auditory pathways. Surrounding this core is a thin shell of microneurons called the granule cell domain (GCD). The GCD receives auditory and nonauditory inputs and projects in turn to the dorsal cochlear nucleus, thus appearing to serve as a central locus for integrating polysensory information and descending feedback. Nevertheless, the source of many of these inputs and the nature of the synaptic connections are relatively unknown. We used the retrograde tracer Fast Blue to demonstrate that a major projection arises from the contralateral pontine nuclei (PN) to the GCD. The projecting cells are more densely located in the ventral and rostral parts of the PN. They also are clustered into a lateral and a medial group. Injections of anterograde tracers into the PN labeled mossy fibers in the contralateral GCD. The terminals are confined to those parts of the GCD immediately surrounding the ventral cochlear nucleus. There is no PN projection to the dorsal cochlear nucleus. These endings have the form of bouton and mossy fiber endings as revealed by light and electron microscopy. The PN represent a key station between the cerebral and cerebellar cortices, so the pontocochlear nucleus projection emerges as a significant source of highly processed information that is introduced into the early stages of the auditory pathway. The cerebropontocerebellar pathway may impart coordination and timing cues to the motor system. In an analogous way, perhaps the cerebropontocochlear nucleus projection endows the auditory system with a timing mechanism for extracting temporal information. PMID- 11438932 TI - Central anatomy of individual rapidly adapting low-threshold mechanoreceptors innervating the "hairy" skin of newborn mice: early maturation of hair follicle afferents. AB - Adult skin sensory neurons exhibit characteristic projection patterns in the dorsal horn of the spinal gray matter that are tightly correlated with modality. However, little is known about how these patterns come about during the ontogeny of the distinct subclasses of skin sensory neurons. To this end, we have developed an intact ex vivo somatosensory system preparation in neonatal mice, allowing single, physiologically identified cutaneous afferents to be iontophoretically injected with Neurobiotin for subsequent histological analyses. The present report, centered on rapidly adapting mechanoreceptors, represents the first study of the central projections of identified skin sensory neurons in neonatal animals. Cutaneous afferents exhibiting rapidly adapting responses to sustained natural stimuli were encountered as early as recordings were made. Well stained representatives of coarse (tylotrich and guard) and fine-diameter (down) hair follicle afferents, along with a putative Pacinian corpuscle afferent, were recovered from 2-7-day-old neonates. All were characterized by narrow, uninflected somal action potentials and generally low mechanical thresholds, and many could be activated via deflection of recently erupted hairs. The central collaterals of hair follicle afferents formed recurrent, flame-shaped arbors that were essentially miniaturized replicas of their adult counterparts, with identical laminar terminations. The terminal arbors of down hair afferents, previously undescribed in rodents, were distinct and consistently occupied a more superficial position than tylotrich and guard hair afferents. Nevertheless, the former extended no higher than the middle of the incipient substantia gelatinosa, leaving a clear gap more dorsally. In all major respects, therefore, hair follicle afferents display the same laminar specificity in neonates as they do in adults. The widely held misperception that their collaterals extend exuberant projections into pain-specific regions of the dorsal horn during early postnatal life is shown to have multiple, deep-rooted underpinnings. PMID- 11438933 TI - Quantitative assessment of retinoid signaling pathways in the developing eye and retina of the chicken embryo. AB - Retinoid signaling has been implicated as an important regulator of retinal development and differentiation. We have used state of the art high-pressure liquid chromatography to identify and quantitate biologically active retinoids, immunohistochemistry to localize the retinoic acid synthetic enzyme retinaldehyde dehydrogenase 2 (RALDH2), and nucleic acid assays to quantitate and localize retinoid receptor gene transcripts in the developing eye and retina of the chicken. Our results demonstrate spatial distinctions in retinoid synthesis and signaling that may be related to laminar differentiation in the developing retina. Retinoic acids (RAs) and their precursor retinols (ROHs) are the predominant retinoids in the developing eye. All-trans-RA and all-trans-3,4 didehydro-RA are present in the neuroepithelium in approximately equal amounts from early stages of neurogenesis until shortly before hatching. The retinoid X receptor (RXR) ligand 9-cis-RA is undetectable at all stages; if present, it cannot exceed a small percentage of the total RA content. RAs are not detected in the pigment epithelium. All-trans-ROH is present in the neuroepithelium and pigment epithelium, whereas all-trans-3,4-didehydro-ROH is detected only in the pigment epithelium and/or the choroid and sclera. RALDH2 immunoreactivity is intense in the choroid, low or absent in the pigment epithelium, and moderate in the neuroepithelium, where it is highest in the outer layers. Transcripts of all five chicken retinoid receptor genes are present in the neural retina and eye throughout development. During the period of neurogenesis, at least three of the receptors (RAR gamma, RXR gamma, RXRalpha), exhibit dynamic patterns of differential localization within the depths of the neural retina. PMID- 11438934 TI - Rod pathways in the mammalian retina use connexin 36. AB - Many neurons in the mammalian retina are coupled by means of gap junctions. Here, we show that, in rabbit retina, an antibody to connexin 36 heavily labels processes of AII amacrine cells, a critical interneuron in the rod pathway. Image analysis indicates that Cx36 is primarily located at dendritic crossings between overlapping AII amacrine cells. This finding suggests that Cx36 participates in homotypic gap junctions between pairs of AII amacrine cells. Cx36 was also found at AII/cone bipolar contacts, previously shown to be gap junction sites. This finding suggests that Cx36 participates at gap junctions that may be heterotypic. These results place an identified neuronal connexin in the context of a well defined retinal circuit. The absence of Cx36 in many other neurons known to be coupled suggests the presence of additional unidentified connexins in mammalian neurons. Conversely, Cx36 labeling in other regions of the retina is not associated with AII amacrine cells, indicating some other cell types use Cx36. PMID- 11438935 TI - Variability of position of the P2 glomerulus within a map of the mouse olfactory bulb. AB - Olfactory receptor neurons (ORNs) that express a common odorant receptor molecule target specific glomeruli in the olfactory bulb. We systematically assessed the location of the olfactory glomeruli that receive input from ORNs expressing P2 receptors in the P2-internal ribosome entry site-tau-lacZ mouse. We present a new mapping method that includes an Internet-accessible computer program for generating two- and three-dimensional maps of the glomerular sheet in the olfactory bulbs of mice. Cylindrical coordinates were used to define glomerular location: The coordinates were given as the anteroposterior (AP) distance parallel to the long axis of the bulb (rostrocaudal; RC) and angular measurements with origin defined by the remnant ependymal layer in the center of the granule cell layer in the bulb. Using this method, we can apply rigorous statistical methods to give objective estimates of position and variability. At the 95% confidence interval, the lateral P2 glomerulus lies at coordinates 1,008 microm +/- 306 microm AP x 146 degrees +/- 12 degrees, and the medial P2 glomerulus lies at 1,828 microm +/- 196 microm AP x 204 degrees +/- 8 degrees. We estimate that these coordinates encompass a domain containing 29 and 37 of the 1,800 glomeruli ( approximately 2%) for the lateral and medial glomeruli, respectively. Furthermore, the data reported here demonstrate that the rostrocaudal position of small P2 glomeruli is three times more variable than that of large glomeruli. PMID- 11438936 TI - Peptidergic neurohemal system of an insect: mass spectrometric morphology. AB - Neuropeptides are by far the most diverse group of messenger molecules in insects. To understand cell signaling and function, it is essential to reveal the complete neuropeptide profile of a single neuron/nerve/neurohemal organ first. In this study, matrix-assisted laser desorption ionization time of flight (MALDI TOF) mass spectrometry was used to analyze the peptidergic system of an insect, focussing on the neurohemal structures. Major neurohemal organs were investigated, including the retrocerebral complex, perisympathetic organs, and all nerves supplying these organs with neurosecretions. Additionally, peripheral neurohemal release sites such as the dilator muscle of the antennal circulatory organ and lateral heart nerves were studied, as well as parts of the stomatogastric nervous system. The following neuropeptide families were analyzed: kinins, allatostatins, leucomyosuppressin, corazonin, adipokinetic hormones, myoinhibitory peptide, sulfakinins, periviscerokinins, YLSamide, VEAacid, SKNacid, proctolin, the head peptide, and pyrokinins. Beyond a contribution to a map of the distribution of neuropeptides in a neurohemal system, the following conclusions can be drawn from these experiments. (1) Nearly all abundant peaks in the different mass spectra represent peptides that have already been identified. (2) Although only adult males were used in this study, variations in the peptide abundances were observed that are possibly correlated with different physiological/developmental conditions. (3) Peptides have a body-region-specific distribution in the neurohemal system. (4) A clear compartmentalisation of the retrocerebral complex could be observed. PMID- 11438937 TI - Localization of neuropeptide Y mRNA and peptide in the chicken hypothalamus and their alterations after food deprivation, dehydration, and castration. AB - Localization of neuropeptide Y (NPY) mRNA in the hypothalamus of chickens was studied by in situ hybridization with digoxigenin-labeled chicken NPY cRNA probe. The largest number of perikarya-expressing NPY mRNA was found within the mediobasal hypothalamus, including the infundibular nucleus, inferior hypothalamic nucleus, and median eminence. Many NPY perikarya were noted to surround the nucleus rotundus and to be present in the supraoptic nucleus. Moreover, some perikarya were detected in the nucleus of basal optic root, bed nucleus pallial commissure, and nucleus striae terminalis close to the lateral forebrain bundle. NPY-immunoreactive nerve fibers were densely distributed in these regions containing the NPY mRNA-expressing perikarya. Following food deprivation for four days, perikarya-expressing NPY mRNA and peptide were markedly increased in the mediobasal hypothalamus and particularly so in the infundibular nucleus. No changes, however, were detected in other regions containing NPY-positive perikarya. Water deprivation induced less increase in NPY positive perikarya in the mediobasal hypothalamus compared to food deprivation. After gonadectomy, the number of NPY-positive perikarya in the mediobasal hypothalamus was unaltered. Northern blot analysis with (32)P-labeled chicken NPY cDNA probe demonstrated that a 2.7-fold increase of NPY mRNA was induced by starvation and a 1.5-fold increase was induced by dehydration, whereas the NPY mRNA band remained unchanged after gonadectomy. Thus, it seems that NPY neurons located in the mediobasal hypothalamus are involved in feeding behavior but not reproductive activity. PMID- 11438938 TI - Heterogeneity of cycling glial progenitors in the adult mammalian cortex and white matter. AB - The heterogeneity and differentiation potential of mitotically active cells in the adult brain were studied by labeling adult rats with BrdU, and isolating an enriched population of cycling cells from neocortex and from subcortical white matter. The majority of this population isolated from either brain region labeled with O4, an early oligodendrocyte marker. In tissue culture, these O4(+) progenitors acquired galactocerebroside, a glycolipid of mature oligodendrocytes, but not GFAP, an intermediate filament of astrocytes. A minority population expressed the intermediate filament protein, vimentin, but not O4. This population expressed GFAP after several days in culture. A third population of cycling cells, expressing the gangliosides labeled with the A2B5 antibody, represented a minority population in subcortical white matter, but one of the major cycling populations in cortex, with substantial overlap with O4. Small populations of cycling NG2(+) cells also were observed. Thus, the cycling cells in the adult brain are heterogeneous, and the majority appear to belong to glial lineages. PMID- 11438939 TI - Neuroprotective signal transduction in model motor neurons exposed to thrombin: G protein modulation effects on neurite outgrowth, Ca(2+) mobilization, and apoptosis. AB - Thrombin, the ultimate protease in the blood coagulation cascade, mediates its known cellular effects by unique proteolytic activation of G-protein-coupled protease-activated receptors (PARs), such as PAR1, PAR3, and PAR4, and a "tethered ligand" mechanism. PAR1 is variably expressed in subpopulations of neurons and largely determines thrombin's effects on morphology, calcium mobilization, and caspase-mediated apoptosis. In spinal cord motoneurons, PAR1 expression correlates with transient thrombin-mediated [Ca(2+)](i) flux, receptor cleavage, and elevation of rest [Ca(2+)](i) activating intracellular proteases. At nanomolar concentrations, thrombin retracts neurites via PAR1 activation of the monomeric, 21 kDa Ras G-protein RhoA, which is also involved in neuroprotection at lower thrombin concentrations. Such results suggest potential downstream targets for thrombin's injurious effects. Consequently, we employed several G-protein-specific modulators prior to thrombin exposure in an attempt to uncouple both heterotrimeric and monomeric G-proteins from motoneuronal PAR1. Cholera toxin, stimulating Gs, and lovastatin, which blocks isoprenylation of Rho, reduced thrombin-induced calcium mobilization. In contrast, pertussis toxin and mastoparan, inhibiting or stimulating G(o)/G(i), were found to exacerbate thrombin action. Effects on neuronal rounding and apoptosis were also detected, suggesting therapeutic utility may result from interference with downstream components of thrombin signaling pathways in human motor neuron disorders, and possibly other neurodegenerative diseases. Published 2001 John Wiley & Sons, Inc. PMID- 11438940 TI - Screening for synaptic defects revealed a locus involved in presynaptic and postsynaptic functions in Drosophila embryos. AB - To identify genes involved in synaptic functions, we screened lethal enhancer trap lines by monitoring synaptic activities at the neuromuscular junction in Drosophila embryos. It was found that MY7919, thus isolated, has moderate defects in both pre- and postsynaptic functions. The mean amplitudes of spontaneous as well as evoked synaptic currents were smaller than those in wild-type. The failure rate was higher than normal at any given concentration of external Ca(2+), indicating that presynaptic functions were impaired. In addition, the mean amplitude of miniature synaptic currents was smaller, and the unitary current amplitudes of junctional glutamate receptor channels were slightly but significantly smaller. Thus, postsynaptic functions were also altered. The gene was cloned and found to be identical to the previously reported apontic (=tracheae defective) locus, which is believed to be a transcription factor expressed in the central nervous system (CNS) as well as in the head, tracheae, and heart. Immunohistochemical analysis using an antiapontic antibody revealed that the protein is localized to nuclei. Null alleles of the apontic locus were obtained by imprecise excision of the enhancer trap vector. Synaptic activities in null mutants were not different from those of the original allele, even though null homozygotes had uncontracted ventral nerve cords and more severe behavioral phenotypes. The morphology of the neuromuscular junction of the null mutant was qualitatively similar to that of wild-type, with the presence of typical pre- and postsynaptic specializations, but with some suggestions of quantitative differences. This strategy for screening mutants with synaptic defects will reveal more genes directly or indirectly affecting synaptic transmission. PMID- 11438941 TI - Dendritic growth induced by BMP-7 requires Smad1 and proteasome activity. AB - Bone morphogenetic proteins (BMPs) induce dendritic growth in cultured sympathetic neurons; however, the signaling pathways that mediate this dendrite promoting activity have not been previously characterized. Here we report studies of the signaling events that regulate the growth of these afferent processes. We find that Smad1 is expressed in sympathetic neurons and that BMPs rapidly induce its phosphorylation and translocation from the cytoplasm to the nucleus. Furthermore, a dominant negative form of Smad1 inhibits BMP-7-induced dendritic growth, suggesting a requirement for Smad1 activation in this biological activity of BMP-7. A physical interaction between Smad1 and components involved in the proteasome-mediated degradation system was detected with a yeast two-hybrid screen, thereby prompting an examination of the effects of proteasome inhibitors on dendritic growth. Lactacystin and ALLN (N-acetyl-Leu-Leu-norleucinal) selectively blocked BMP-7-induced dendritic growth without adversely affecting either cell viability or axonal growth. Moreover, studies of transfected P19 cells suggest that the proteasome inhibitors directly block the effects of Smad1 on the transcriptional activity of the Tlx-2 promoter. These data indicate that BMP-induced dendritic growth requires Smad1 activation and involves proteasome mediated degradation events. PMID- 11438942 TI - Neurotrophins, but not depolarization, regulate substance P expression in the developing optic tectum. AB - Neurotransmitter expression can be regulated by both activity and neurotrophins in a number of in vitro systems. We examined whether either of these factors was likely to play a role in the in vivo optic nerve-dependent regulation of a substance P-like immunoreactive (SP-ir) population of cells in the developing optic tectum of the frog. In contrast to our previous results with the adult system, blocking tectal cell responses to glutamate release by retinal ganglion cells with 6-cyano-7-nitroquinoxaline-2,3 dione (CNQX) did not affect the percent of SP-ir cells in the developing tectum. Treatment with d-(-)-2-amino-5 phosphonovaleric acid (d-AP-5) was also ineffective in this regard, although both it and CNQX treatment disrupted visual map topography. Chronic treatment with brain-derived neurotrophic factor (BDNF) and neurotrophin-4/5 (NT-4/5) produced increases in SP-ir cells in the treated lobes of normal animals, which were significant in the case of NT-4/5. Both substances also prevented the decrease of SP cells that would otherwise occur in the deafferented lobe of unilaterally optic nerve-transected tadpoles. These changes in the percent of SP-ir cells occurred without any detectable changes in the overall number of tectal cells. NGF had no effect on SP expression. Nor did it affect topographic map formation, which was disrupted by treatment with either BDNF or NT-4/5. Our results demonstrate that different mechanisms regulate SP expression in the developing and adult tectum. They indicate that neurotrophin levels in the developing optic tectum may selectively regulate a specific neuropeptide-expressing population of cells. PMID- 11438943 TI - Cell-type specific dendritic contacts between retinal ganglion cells during development. AB - The extent of a neuron's dendritic field defines the region within which information is processed. The dendritic fields of functionally distinct ON and OFF center retinal ganglion cells (RGCs) form separate mosaics across the retina. Within each mosaic, neighboring dendritic fields overlap by a constant amount, sampling the visual field with the appropriate coverage. Contact-mediated lateral inhibition between neighboring RGCs has long been thought to regulate both the extent and overlap of dendritic fields during development. Here we show that dendro-dendritic contact exists between developing RGCs and occurs in a manner that would regulate the formation of ON and OFF mosaics separately. Dye-filled neighboring ON and OFF ferret alpha RGCs were reconstructed using multiphoton microscopy. At all neonatal ages examined, we observed dendro-dendritic contacts between RGCs of the same sign (ON/ON; OFF/OFF), but never between cells of opposite signs (ON/OFF). Terminal dendrites of one cell often touched a dendrite of its neighbor as they intersected. In some instances, the distal dendrite of one cell formed a fascicle with the proximal process of its neighbor. Alpha cells did not form contacts with neighboring beta cells of the same sign. Together, these observations suggest that dendro-dendritic contact between RGCs is cell type specific. Dendritic contacts were observed even before the alpha cell arbors were completely stratified, suggesting that cell-cell recognition may take place early in their development. For each cell type, the relative overlap of dendritic fields was constant with age, despite a two-fold increase in field area. We suggest that dendro-dendritic contacts may be sites of intercellular signaling that could regulate local extension of dendrites to maintain the relative overlap of RGCs within a mosaic during development. PMID- 11438944 TI - Prenatal diagnosis of complete sole trisomy 1q. AB - The prenatal diagnosis of a complete trisomy of the long arm of chromosome 1 is reported. Major ultrasound findings included: nuchal thickening, bi-temporal narrowing, a single choroid plexus cyst, and mild ventriculomegaly. There was a mass in the chest and abdomen, pleural effusion, ascites and a hyperechoic bowel. Skin edema was present. The fetus died at 26 weeks' gestation. A literature review is presented of 17 de novo and two inherited cases with only trisomy 1q. Of note is the fact that 3/5 prenatally detected 1q trisomies have teratomas. A review of the literature reveals a dismal outcome for trisomy 1q cases if the duplication involves bands 1q25-->q32. PMID- 11438945 TI - Maternal serum levels of dimeric inhibin A in pregnancies affected by trisomy 21 in the first trimester. AB - Dimeric inhibin A was measured in maternal serum samples from 45 pregnancies affected by trisomy 21 and 493 samples from unaffected pregnancies at 10-14 weeks of gestation. Inhibin A levels in affected pregnancies were compared with levels of free beta-hCG and PAPP-A in the same series. In the trisomy 21 group, the median multiple of the median (MoM) inhibin A was not significantly elevated (1.28 vs 1.00) with only 15.5% being above the 95th centile. In contrast, the median MoM free beta-hCG was significantly increased (2.05 vs 1.00) with 36% above the 95th centile and PAPP-A was significantly reduced (0.49 vs 1.00) with 42% below the 5th centile. Inhibin A levels in the trisomy 21 group were significantly correlated with gestational age such that median levels rose from 1.04 at 11 weeks to 1.30 at 12 weeks and 1.67 at 13 weeks. These findings suggest that first trimester biochemical screening for trisomy 21, which is currently optimised using maternal serum free beta-hCG and PAPP-A and fetal nuchal translucency, will not benefit from the inclusion of inhibin A. PMID- 11438946 TI - Between pregnancy biological variability of first trimester markers of Down syndrome: implications for screening in subsequent pregnancies. AB - In a group of 149 women who had undergone routine first trimester screening using fetal nuchal translucency thickness (NT) and maternal serum free beta-hCG and pregnancy associated plasma protein-A (PAPP-A) in two consecutive pregnancies the within person between pregnancy biological variability of these markers has been assessed. For fetal NT there was no correlation between NT MoM in the first and second pregnancy (r=0.0800). For maternal serum free beta-hCG MoM a significant correlation was observed (r=0.4174) as was also found for PAPP-A MoM (r=0.3270). The implications for such between pregnancy marker association is that women who have an increased risk of Down syndrome in their first pregnancy are 1.5-2 times more likely to repeat this event in their next pregnancy. This observation may be useful in counselling women in the first trimester screening of a subsequent pregnancy. PMID- 11438947 TI - Attitude of Saudi families affected with hemoglobinopathies towards prenatal screening and abortion and the influence of religious ruling (Fatwa). AB - Hemoglobinopathies are common inherited disorders in Saudi Arabia. Prenatal diagnosis for such diseases is specific and sensitive but not yet implemented in Saudi Arabia. Saudis are Muslims with a very high rate of consanguinity and inherited genetic disorders. To examine the attitude of Saudi families affected with hemoglobinopathies towards prenatal diagnosis and abortion, and to evaluate the effect of education on religious ruling on such attitudes, 32 families were interviewed using a pre-structured questionnaire. The majority accepted prenatal diagnosis (81.3%). The attitude towards abortion was greatly affected by religious values. Education about religious ruling significantly affected parents' attitude towards accepting abortion and prenatal diagnosis. No other factors were found to influence the outcome. Although the majority of families received some kind of formal genetic counseling [23/32 (71.9%)], none of them was informed about the possibility of prenatal or preimplantation diagnosis prior to the interview. Therefore for prevention of genetic disorders, the emphasis in countries with a vast majority of Muslims such as Saudi Arabia has probably to be placed on public awareness about genetic risks, the risk of consanguinity, availability of services, and so on, while at the same time taking into consideration the religious beliefs and education of the target population PMID- 11438948 TI - Pulmonary stenosis and reactive right ventricular hypertrophy in the recipient fetus as a consequence of twin-to-twin transfusion. AB - The present study describes an association between adverse outcome in the twin-to twin transfusion syndrome (TTTS) and pulmonary stenosis or reactive right ventricular hypertrophy. Six discordant monozygotic twin pregnancies with TTTS are described. Ventricular hypertrophy and atrioventricular valvular regurgitation occurred in all the recipient twins with pulmonary valvular stenosis in three cases and infundibular stenosis in one case. The recipient twin in one pair and both twins in another pregnancy died as a consequence of immaturity but the remaining twins all survived. Surgical intervention was required in one baby for valvular pulmonary stenosis. Our observations suggest that elevated blood pressure in the transfusion recipient may play an important role in pathogenesis. We hypothesise that both pulmonary stenosis and right chamber hypertrophy are secondary to hemodynamic changes. Although we have found valvular pulmonary stenosis in three recipients and infundibular stenosis in only one, this (obstruction to outflow) could be due to right chamber hypertrophy. PMID- 11438949 TI - Prenatal diagnosis of mosaicism for triploidy and trisomy 13. AB - Mosaicism for trisomy 13 and triploidy was detected by amniocentesis performed at 18 weeks' gestation because of fetal anomalies. Pregnancy continued and a live born male was delivered vaginally at 37 weeks. The infant had features common to both trisomy 13 and triploidy: intrauterine growth retardation (IUGR), small abnormal ears, cleft palate, and a small jaw. In addition, he had complete cutaneous syndactyly of fingers 3 and 4 and partial syndactyly of the toes, as seen in triploidy. Mixoploidy for trisomy 13 and triploidy was confirmed postnatally in blood, skin, and placenta. Examination of chromosome heteromorphisms and DNA markers suggested the presence of two maternal contributions in the triploid cell line. In addition, the extra chromosome 13 in the trisomic cell line was derived from the mother. PMID- 11438950 TI - Prenatal diagnosis of sex chromosome aneuploidy: possible reasons for high rates of pregnancy termination. AB - Sex chromosome aneuploidy (SCA), when detected in amniocentesis, is usually an unexpected result of a test carried out for another purpose. For most SCAs, the prognosis is milder and less predictable than trisomy 21, and therefore parents are faced with a difficult decision regarding the option of pregnancy termination. While studies from Europe and the USA report a declining trend in termination rates for SCA, our local experience is different. During the period 1989-1998, we diagnosed 60 SCA (including mosaics) in 20 106 amniocenteses (0.29%) and 48 (80%) of these pregnancies were terminated, a significantly higher proportion than has been reported in Europe and the USA. The present study shows that the difference between our experience and others' may be related to differences in cultural norms and values. Thirty women were interviewed, of whom 23 terminated and seven continued the pregnancy. Interview analyses showed that the main reason behind the decision to terminate the pregnancy was associated with the parents' fear of non-specific abnormality of the child, and concerns about abnormal sexual development. Although genetic counseling practised in our center aims to be non-directive, 56% of the women reported that the counseling was either directive towards termination, or that they at least felt that the counselor's attitude was pro-termination. Most women (93%) reported themselves as having come to terms with their decision. PMID- 11438951 TI - Familial orofaciodigital syndrome type I revealed by ultrasound prenatal diagnosis of porencephaly. AB - Porencephaly is a rare central nervous system (CNS) abnormality that can be caused by an intraparenchymal destructive process or a developmental defect. Here we report on a prenatal ultrasound diagnosis of complex CNS abnormalities including agenesis of the corpus callosum, agenesis of the cerebellar vermis, bilateral hydrocephaly, and bilateral porencephaly in fetus at 33 weeks' gestation. The diagnosis of familial orofaciodigital syndrome type I (OFD I) was raised after fetal autopsy, clinical examination of the family, and the X-linked dominant inheritance pattern. This is the fourth report of porencephaly in association with OFD I. We discuss the difficulties in genetic counselling since OFD I shows variable expressivity of the phenotypic features. Furthermore, we emphasize the importance of a detailed ultrasound examination after a prenatal diagnosis of porencephaly. PMID- 11438952 TI - Rapid and radical amniodrainage in the treatment of severe twin-twin transfusion syndrome. AB - We have evaluated the role of a rapid and radical method of amniodrainage in the treatment of severe twin-twin transfusion. The outcome of 15 patients with severe twin-twin transfusion for which a amniodrainage was performed by means of a vacuum bottle system was compared with the outcome of 15 patients with a similar condition, matched for gestational age at the time of the initial procedure and drained using a standard procedure. In the study group the amniodrainage ended when no amniotic fluid could be aspirated, whereas the women in the standard group were drained with a syringe system and the fluid was removed until the deepest amniotic fluid pool was <8 cm. At the initial procedure, the mean volume of amniotic fluid drained was significantly (p<0.05) higher (3252 vs 2153 ml) and the length of the procedure significantly (p<0.001) shorter (21 vs 41 min) in the study group than in the standard group. The mean post-procedure amniotic fluid index was significantly (p<0.001) smaller (2.9 vs 7.7 cm) after radical amniodrainage than after the standard amniodrainage. The mean number of procedures was significantly (p<0.001) lower (1.5 vs 5.6) in the study group compared to the standard group. In the study group the mean placental thickness increased significantly (p<0.001) from 9 mm before the procedure to 49 mm after, and the overall perinatal survival rate was 80% and the proportion of pregnancies with at least one survivor was 93%. The present data indicate that early, rapid and radical amniodrainage is an effective and low-cost therapy for severe twin twin transfusion syndrome. Compared to the standard amniodrainage technique it also appears to reduce the need for multiple procedures. PMID- 11438953 TI - Prenatal diagnosis of a de novo supernumerary marker derived from chromosome 16. AB - Marker chromosomes are supernumerary chromosomes of unknown origin and are seldom found in prenatal diagnosis. Application of fluorescent in situ hybridization (FISH) allows the identification of the chromosomal origin of markers. Estimation of the risk of an abnormal phenotype outcome can be enabled by collecting data on phenotypes associated with markers of the same chromosomal origin. So far only very few cases of prenatal diagnosis of de novo supernumerary markers derived from chromosome 16 have been reported. Here the prenatal diagnosis of a de novo supernumerary marker chromosome 16 is described and the relevant literature discussed. PMID- 11438954 TI - Complete karyotype discrepancy between placental and fetal cells in a case of ring chromosome 18. AB - A case of complete karyotype discrepancy between cultured chorionic villi and amniotic in addition to fetal cells is reported. Ring chromosome 18 and monosomy 18 mosaicism was detected after amniocentesis. The pregnancy was terminated in the 23rd gestational week. Cytogenetic analysis of cultured umbilical cord tissue after termination confirmed the finding of ring chromosome 18/monosomy 18 mosaicism. In cultured umbilical blood lymphocytes monosomic cells 45,-18 were not detected and the karyotype was 46,XY,r(18). In contrast, short-term and long term cultured chorionic villi showed a normal male karyotype of 46,XY. Ultrasonographic examination revealed amniotic band syndrome and scoliosis in the caudal region of the spine. PMID- 11438955 TI - Prenatal diagnosis of two rare de novo structural aberrations of the Y chromosome: cytogenetic and molecular analysis. AB - Two rare de novo structural aberrations of the Y chromosome were detected during routine prenatal diagnosis: a satellited non-fluorescent Y chromosome (Yqs), the first de novo Yqs to be reported in a fetus, and a terminal deletion of the Y chromosome long arm del(Y)(q11). In both cases detailed cytogenetic and molecular analyses were undertaken. In the case of the Yqs it was demonstrated by fluorescence in situ hybridization (FISH) that the satellites were derived from chromosome 15. In the case of the del(Yq), it was shown with molecular analysis by polymerase chain reaction (PCR) amplification of sequence-tagged sites (STS PCR) that the deleted portion of the long arm of chromosome Y included the azoospermia factor loci, AZFb and AZFc. The clinical significance of these findings is discussed. PMID- 11438956 TI - A comparison of different lysis buffers to assess allele dropout from single cells for preimplantation genetic diagnosis. AB - Single cell polymerase chain reaction (PCR) for preimplantation genetic diagnosis (PGD) requires high efficiency and accuracy. Allele dropout (ADO), the random amplification failure of one of the two parental alleles, remains the most significant problem in PCR-based PGD testing since it can result in serious misdiagnosis for compound heterozygous or autosomal dominant conditions. A number of different strategies (including the use of lysis buffers to break down the cell and make the DNA accessible) have been employed to combat ADO with varying degrees of success, yet there is still no consensus among PGD centres over which lysis buffer should be used (ESHRE PGD Consortium, 1999). To address this issue, PCR amplification of three genes (CFTR, LAMA3 and PKP1) at different chromosomal loci was investigated. Single lymphocytes from individuals heterozygous for mutations within each of the three genes were collected and lysed in either alkaline lysis buffer (ALB) or proteinase K/SDS lysis buffer (PK). PCR amplification efficiencies were comparable between alkaline lysis and proteinase K lysis for PCR products spanning each of the three mutated loci (DeltaF508 in CFTR 90% vs 88%; R650X in LAMA3 82% vs 78%; and Y71X in PKP1 91% vs 87%). While there was no appreciable difference between ADO rates between the two lysis buffers for the LAMA3 PCR product (25% vs 26%), there were significant differences in ADO rates between ALB and PK for the CFTR PCR product (0% vs 23%) and the PKP1 PCR product (8% vs 56%). Based on these results, we are currently using ALB in preference to PK/SDS buffer for the lysis of cells in clinical PGD. PMID- 11438957 TI - Allele-specific amplification for preimplantation genetic diagnosis (PGD) of spinal muscular atrophy. AB - We have developed a new allele-specific amplification method for the preimplantation genetic diagnosis (PGD) of spinal muscular atrophy (SMA; Werdnig Hoffmann disease) from a single cell. This method is based on the detection of the deletion of exon 7 of the telomeric copy of the survival motor neurone (SMN(t)) gene. An oligonucleotide was designed to be specific to the SMN(t) nucleotidic sequence with exonic mismatch G (for SMN(t))-->A (for SMN(c)) at its 3' end. This test produces reliable PCR products in 95% of single lymphoblasts (85/88) tested as well as in 16/16 blastomeres from normal controls. Specificity analysis showed that we were able to detect homozygous deletion of the SMN(t) gene in 99% of single lymphoblasts (103/104) from a SMA patient. No contamination was detected in 68 blanks tested. Multiple cell and DNA dilution analysis revealed that the test is accurate and specific up to 100 pg DNA and should thus also be suitable for PGD at the blastocyst stage. This rapid procedure requires a single round of fluorescent PCR and no restriction digestion, while previously described single cell methods include nested PCR followed by restriction enzyme digestion. Two PGD cycles for SMA using this procedure were performed in our centre. PMID- 11438958 TI - Preimplantation genetic diagnosis of the fragile X syndrome by use of linked polymorphic markers. AB - Fragile X syndrome is the most common cause of familial mental retardation. The most common mutation is expansion of a triplet (CGG)(n) repeat in the 5' untranslated region of the FMR1 gene on Xq27.3. The expansion is refractory to PCR due to preferential amplification of the smaller allele in heterozygous cells and the high GC content of the repeat and surrounding sequences. Direct detection of the normal parental alleles in preimplantation embryos has been used for preimplantation genetic diagnosis (PGD) of this disorder. However, this approach is only suitable for approximately 63% of couples due to the heterozygosity of the repeat in the normal population. As an alternative we investigated the use of polymorphic markers flanking the mutation to track the normal and premutation carrying maternal chromosomes in preimplantation embryos. Using a panel of 11 polymorphisms, six (CA)(n) repeats and five single nucleotide polymorphisms, diagnosis was developed for 90% of referred couples. Multiplex amplification of informative markers was tested in 300 single buccal cells from interested couples with efficiency and allele drop out (ADO) rates ranging from 69% to 96% and 6% to 18%, respectively. Use of this approach is accurate and applicable to a larger number of patients at risk of transmitting fragile X to their offspring. PMID- 11438959 TI - Prenatal diagnosis of a fetus with megacystis and monosomy 21. PMID- 11438960 TI - Prenatal testing for uniparental disomy (UPD). PMID- 11438962 TI - A study of chiral recognition for NBD-derivatives on a Pirkle-type chiral stationary phase. AB - A chiral stationary phase (CSP 1) derived from an (S)-N-3,5-dinitrobenzoyl-1 naphthylglycine showed excellent enantiomeric separation for amino acid derivatives with a fluorogenic reagent, 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), in high-performance liquid chromatography (HPLC). We compared elution profiles (separation factor and elution order) of NBD-amino acids and their analogs on HPLC, to determine the diastereomeric complex between the chiral moiety of CSP 1 and NBD-amino acid, which is responsible for the chiral recognition. (1)H-NMR studies of a mixture of model compound of CSP 1 and NBD-Ala suggest that the diastereomeric complex is composed of two hydrogen bonding sites at the amino proton and oxygen atom, and a pi-pi interaction by the benzofurazan structure (2,1,3-benzoxadiazole) of NBD-amino acid. Furthermore CSP 1 was able to separate esters, amides and alpha-methyl amino acids derivatized with NBD-F. PMID- 11438963 TI - Determining the procedure for routine residue monitoring of sulfamethazine in edible animal tissues. AB - A simplified method to determine/identify residual sulfamethazine (SMZ) in edible tissues from cattle, pigs, chickens and sheep by a high-performance liquid chromatography (HPLC) with a photo-diode array detector is presented. The sample preparation was performed by homogenizing with 30% (v/v) ethanol in water followed by an Ultrafree-MC/PL as a centrifugal ultra-filtration unit. For determination/identification of SMZ, a reversed-phase C(4) column and a mobile phase of 15% (v/v) ethanol in water with a photo-diode array detector was used. Average recoveries from spiked SMZ (0.1-1.0 mg kg(-1)) were >or=80% with coefficients of variation between 1.3 and 4.3%. The limits of quantitation were calculated to be 0.057-0.060 mg kg(-1). The total time and solvent required for the analysis of one sample were <40 min and <2 mL of ethanol, respectively. PMID- 11438965 TI - Determination of trimebutine maleate in rat plasma and tissues by using capillary zone electrophoresis. AB - A simple and rapid capillary zone electrophoresis method was developed for the determination of trimebutine maleate in rat plasma and tissues. Rat plasma and tissue homogenates were mixed with acetonitrile containing internal standard, ephedrine hydrochloride, and then centrifuged. The supernatant was dried under a stream of nitrogen, and the residue was reconstituted in methanol-water (1:1). The electrophoresis was performed in uncoated capillary with 30 mmol/L phosphate buffer of pH 6.0 as the separation electrolyte. The applied voltage was 10 kV and the UV detection was set at 214 nm. The peak height ratio vs concentration in plasma or homogenates was linear over the range of 5-500 ng/mL and the limit of quantitation was 5 ng/mL. The intra- and inter-day precision was RSD < 14% and <15%. The accuracy was relative error (RE) within +/- 14%. This method was applied to studying the pharmacokinetics and tissue distribution after a single dose of trimebutine maleate was administrated to the rats. The T(max), AUC, C(max) and t(1/2) were 30 min, 7.8 x 10(2) (ng/mL) min, 39 ng/mL and 1.7 x 10(2) min. The drug distribution was found in a decreasing order of liver, kidney, spleen, lung and heart. PMID- 11438964 TI - Determination of rat hepatocellular glutathione by reversed-phase liquid chromatography with fluorescence detection and cytotoxicity evaluation of environmental pollutants based on the concentration change. AB - Three methods for the determination of rat hepatocellular thiols by high performance liquid chromatography (HPLC) with fluorescence (FL) detection have been developed. The thiols in the cells were tagged with three fluorogenic reagents, SBD-F, ABD-F and DBD-F. These reagents could permeate into cells and effectively reacted with thiols to produce highly fluorescent derivatives. These derivatives fluoresced in the long wavelength region at around 530 nm (excitation at around 380 nm). The five biological thiols tagged were perfectly separated by reversed-phase liquid chromatography and were sensitively and selectively detected without any interference from endogenous substanaces. The main thiol in the cells was reduced GSH and the concentration was at the mM level. The proposed procedures were applied to the determination of hepatocellular GSH after treatment of environmental pollutants such as volatile organic compounds (VOC) and endocrine disrupting chemicals (EDC). From the comparison of intracellular GSH concentration, the test compounds were classified into four groups: compounds of strong depletion (eg triphenyltin chloride, hexachlorocyclohexene, nonylphenol, bromoacetic acid, 4-chlorobenzyl chloride and 1,3-dichloropropene), slight decrease (eg bisphenol A, benzo[a]pylene, carbon tetrachloride and benzene), slight increase (eg bromoform and toluene), and no effect (eg 1,1,1 trichloroethane, 1,1,2-trichloroethane and 1,2-dichloroethane). Although the decrease of GSH concentration does not reflect the cytotoxicity of chemicals, the proposed procedure utilizing isolated rat hepatpcytes seems to be useful for investigating the bioactivation of VOC, and EDC, etc. PMID- 11438966 TI - Separation of naturally occurring triterpenoidal saponins by capillary zone electrophoresis. AB - A high-performance capillary electrophoresis (HPCE) was successfully applied to the separation and quantitation of naturally occurring oleanene triterpenoidal saponins. The HPCE adapted to the separation of two pairs of disteriomeric saponins (1-2) or (3-4), obtained from Trifolium alexandrinum seeds, was based on capillary zone electrophoresis (CZE) in borate buffer with UV detection at 195 nm. An usual technique for isolation and group separation of saponins was developed as an appropriate purification step prior to determination of individual saponins by CZE. The separation parameters such as borate concentration, pH and applied voltage were varied in order to find the best compromise that complied with demands for high separation, short duration and sufficiently high detector response. The optimum running conditions were found to be 60 mM borate buffer, pH 10 and 12 kV. Under the alkaline borate electrolyte, no resolution was achieved for the saponins (1 and 3) or (2 and 4) in a single mixture, except when 20 mM beta-cyclodextrin was added to the running electrolyte. With the combined techniques of group separation, purification and CZE, a rapid and efficient method for the determination of naturally occurring diasteriomeric saponins is now available. PMID- 11438967 TI - Ascertainment of D-amino acids in germ-free, gnotobiotic and normal laboratory rats. AB - Free D-amino acids were ascertained in the blood serum, urine and aqueous ethanolic extracts of feces of germ-free laboratory rats and a rat made gnotobiotic (tetra-associated) with species of Streptococcus, Lactobacillus and Clostridium. D-Amino acids were also determined in the brains of two germ-free rats. For comparison, D-amino acids were also measured in the blood serum of normal rats and the blood plasma, urine and feces of normal white mice. D Enantiomers of most protein L-amino acids were detected in all physiological samples of animals. Quantities of free D-amino acids were determined as N(O) pentafluoropropionyl-(2)-propyl esters by enantioselective gas chromatography and mass spectrometry. Stereoisomers of the bacterial marker 2,6-diaminopimelic acid, analyzed as N-trifluoroacetyl-(2)-propyl esters, were detected in feces of the gnotobiotic but not of the germ-free rat. PMID- 11438968 TI - A rapid, sensitive high performance liquid chromatographic method for the determination of meropenem in pharmaceutical dosage form, human serum and urine. AB - A new, simple, precise and rapid high performance liquid chromatographic method was developed for the determination of meropenem in human serum, urine and pharmaceutical dosage forms. Chromatography was carried out on an LC(18) column using a mixture of 15 mM KH(2)PO(4):acetonitrile:methanol (84:12:4; v/v/v), adjusted to pH 2.8 with H(3)PO(4). The proposed method was conducted using a reversed-phase technique, UV monitoring at 307.6 nm and cefepime as an internal standard. The retention times were 5.98 and 7.47 min for cefepime and meropenem, respectively. The detector response was linear over the concentration range of 50 10,000 ng/mL. The detection limit of the procedure was found to be 22 ng/mL. The detection limit for meropenem in human plasma was 108.4 ng/mL and the corresponding value in human urine was 179.3 ng/mL. No interference from endogenous substances in human serum, urine and pharmaceutical preparation was observed. The proposed method is sufficiently sensitive for determination of the concentrations of meropenem and may have clinical application for its monitoring in patients receiving the drug. PMID- 11438969 TI - High performance liquid chromatography analysis of 9-(2',3'-dideoxy-2'beta-fluoro D-threo-penta furanosyl) adenine and its metabolite in human plasma using solid phase extraction on a polyfluorinated reversed stationary phase. AB - A quick and sensitive reversed-phase HPLC method has been developed for the analysis of 2'-beta -fluoro-2',3'-dideoxy adenosine (F-ddA), the acid-stable anti AIDS drug, and its metabolite 2'-fluoro-2',3'-dideoxy inosine (F-ddI) in human plasma using polyfluorinated stationary phase column (Fluo fix, 15 cm, 4.0 mm i.d., 5 microm particle size). The mobile phase consisted of ammonium phosphate buffer solution (10 mM) adjusted with phosphoric acid 85% to pH 6.8:dimethyl formamide (97:3, v/v). F-ddA and F-ddI were monitored by UV-visible detector at 258 and 247 nm, respectively. The recoveries of F-ddA and F-ddI from plasma using a C(18) solid-phase extraction cartridge were 99.2% and 99.7%, respectively. PMID- 11438970 TI - Application of capillary electrophoresis in the analysis of novel synthetic dideoxynucleoside analogues with potential anti-HIV activity. AB - The aim of this study was the development of a capillary electrophoretic method for the analysis of a series of novel synthetic dideoxynucleoside analogues with potential anti-HIV activity. These analogues consist of a tetrahydrofuranyl or a tetrahydropyranyl ring as the pseudosugar part and bear a hydroxyethyl side-chain and a nucleobase of the pyrimidine (eg thymine or uracil) or the purine (adenine) type with cis or trans configuration. Analysis of these derivatives was performed by capillary zone electrophoresis using 25 mM phosphate pH 3.00 and 4.00 as operating buffers for pyrimidine and purine analogues, respectively, and detection of separated species at 254 nm. PMID- 11438971 TI - Quantification of D/L-aspartic acids in Aplysia californica central nervous system by beta-cyclodextrin modified micellar electrokinetic chromatography. AB - In presence of an organic modifier (e.g. methanol), separation of amino acid enantiomers tagged with naphthalene-2,3-dicarboxaldehyde by beta-cyclodextrin modified micellar electrokinetic chromatography was dramatically improved. Coupled with laser-induced fluorescence detection, the method was well suited for analysis of D/L-amino acid enantiomers present in mass/volume-limited biological samples such as cell clusters. The five major ganglia dissected from the central nervous system of Aplysia californica, a widely used neuronal model, were analyzed to determine D- and L-aspartic acid enantiomers both free and bound in proteins/tissue matrix. The analyses revealed high levels of free D-aspartic acid ranging from 0.13 to 0.82 micromol/g wet tissue (or 6.0-21.2% of the total free aspartic acid) in all of the five ganglia. However, no D-aspartic acid was detected bound in protein/tissue matrix. The content of free D-aspartic acid in the liver tissue was also found below the detection limit of the method, which was 1 x 10(-8) M. PMID- 11438972 TI - Simultaneous determination of clozapine, norclozapine and clozapine-N-oxide in human plasma by high-performance liquid chromatography with ultraviolet detection. AB - A reversed-phase high-performance liquid chromatographic (HPLC) method for the simultaneous determination of clozapine and its two major metabolites, norclozapine and clozapine-N-oxide in human plasma has been developed and validated. The isocratic HPLC assay uses a mobile phase consisting of an acetonitril-buffered aqueous solution containing 146 microL of triethylamine and 200 microL of 85% phosphoric acid, adjusted to pH 3.3 with 10% potassiumhydroxide solution (400:600, v/v) at a flow-rate of 0.8 ml/min and a Lichrospher 100 RP-18 reversed-phase column and UV detection at 215 nm. Doxepine was used as the internal standard. Mean recoveries for clozapine, norclozapine, clozapine-N-oxide and doxepine were 95%, 98%, 96% and 94%, respectively, whereas the respective mean repeatability coefficients of variation were 3.4%, 2.7%, 4.3% and 0.9%. Reproducibility coefficients of variation were 1.3%, 1.8%, 3.6% and 0.5%, respectively. The mean correlation coefficient for the linear calibration curve (n = 2) for clozapine and norclozapine at a concentration range of 100-1600 ng/mL was 0.9998 and 0.9997, respectively; for clozapine-N-oxide (20-200 ng/mL) it was found to be 0.9986. The lower limits of quantitation were 12.5 ng/mL, 10 ng/mL and 12.5 ng/mL for clozapine, norclozapine and clozapine-N-oxide, respectively. PMID- 11438973 TI - Determination of malondialdehyde by capillary electrophoresis, application to human plasma and relation of its levels with prematurity. AB - Malondialdehyde (MDA) is considered as the most important marker for monitoring lipid peroxidation, which is strongly associated with the development of serious diseases in adults and premature neonates. In this paper we report a method for determination of free MDA in human plasma using capillary zone electrophoresis. MDA was separated and determined as conjugate with tetrabutylammonium hydrogen sulphate (TBAS). Analysis was performed using 20 mM borare, pH = 9.3, as operating buffer and detection of the MDA-TBAS adduct at 267 nm. The method has a linear range up to 80 microM with a detection limit of 0.2 microM. The method was applied to the analysis of MDA in plasma of healthy adults, normal-gestation infants and of preterm neonates. Plasma proteins were successfully removed following centrifugation through a centricon-3 membrane. Results showed that the method can be easily and accurate applied for the determination of MDA in human plasma and that the level of MDA in pretern neonates is significantly higher (p A, a 4 bp insertion 504insAACG, and a 3 bp deletion 358delAGG in three unrelated patients. In 120 patients with normal Cx26 sequence, DHPLC was normal. These results yield sensitivity and specificity of 100% for DHPLC-based detection of Cx26 mutations, and demonstrate that DHPLC is a highly sensitive and specific method of screening for sequence variations in Cx26 that is time and labor efficient. Further, our experience suggests that DHPLC screening alone followed by DNA sequencing only when DHPLC is abnormal may be adequate for identification of all sequence alterations in Cx26. PMID- 11438993 TI - Eight novel ABCD1 gene mutations and three polymorphisms in patients with X linked adrenoleukodystrophy: The first polymorphism causing an amino acid exchange. AB - X-ALD is a neurological disorder associated with inherited defects in the ABCD1 (ALD) gene located on Xq28 and with impaired peroxisomal very long-chain fatty acid beta-oxidation. We examined the ABCD1 gene in probands from 11 unrelated X ALD Czech and Slovak families by the direct sequencing of cDNA or genomic PCR products. In 10 families there were 10 different mutations, eight of which were novel. The spectrum of mutations consists of six point mutations, three microdeletions (1bp, 2bp, 4 bp), and one large deletion (229bp). In the 11th family we detected two novel single-base pair substitutions in exon 1 (c.38 A>C and c.649 A>G), both causing amino acid exchanges (N13T and K217E). Expression studies revealed that only K217E is a deleterious mutation, because a plasmid encoding ALDP with K217E was ineffective in the restoration of defective beta oxidation in X-ALD fibroblasts. The N13T amino acid exchange, on the other hand, did not affect ALDP function. Thus, N13T represents the first polymorphism causing an amino acid exchange in the ABCD1 gene. As this polymorphism was observed neither in 100 control alleles nor in 300 X-ALD patients who have been sequenced so far world-wide, it seems to be very rare or unique. Two additional novel polymorphisms were found by the sequencing of the ABCD1 gene from our patients: c.-59 C/T in the 5'untranslated region and c.2019 C/T (F673F) in exon 10. The frequencies of these two polymorphisms, were 11/150 and 2/150 control alleles, respectively. PMID- 11438994 TI - Haplotypic determinants of instability in the FRAX region: Concatenated mutation or founder effect? AB - The fragile X triplet repeat expansion at Xq27.3 has been shown to be associated with mutation or instability 600 kb distal at the FMR2 repeat locus. Concatenated mutation, whereby a mutation at one locus somehow interacts with mutation, recombination, deletion, or transposition at another locus, is a possible explanation. In this study we examine evidence from a sample of over 7,000 independent haplotypes from the FRAX region. We adopt the use of cladistic groups to more thoroughly define the properties of these haplotypes, and in doing so isolate one group of haplotypes which may be predisposed to the phenomenon of concatenated mutation. Distinguishing concatenated mutation from founder effects is difficult within a single population. We present our evidence for and against concatenated mutation, and in the process describe a previously undefined mutation at FRAXE. PMID- 11438995 TI - Biosensor technology for real-time detection of the cystic fibrosis W1282X mutation in CFTR. AB - In the present paper, biospecific interaction analysis (BIA) was performed using surface plasmon resonance (SPR) and biosensor technologies to detect the Trp1282Ter mutation (W1282X) of the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) gene. We first immobilized on a SA5 sensor chip a single-stranded biotinylated oligonucleotide containing the sequence involved in this mutation, and the efficiency of hybridization of oligonucleotide probes differing in length was determined. Second, we immobilized on different SA5 sensor chips biotinylated polymerase-chain reaction (PCR) products from a normal subject as well as from heterozygous and homozygous W1282X samples. The results obtained show that both allele-specific 10- and 12-mer oligonucleotides are suitable probes to detect W1282X mutations of the cystic fibrosis gene under standard BIA experimental conditions. During the association phase performed at 25 degrees C, discrimination between mismatched and full matched hybrids was readily and reproducibly observed by using the 10-mer W1282X probes. By contrast, when the 12-mer DNA probes were employed, discrimination between mismatched and full matched hybrids was observed during the dissociation phase. Taken together, the results presented suggest that BIA is an easy, speedy, and automatable approach to detect point mutations leading to cystic fibrosis. By this procedure, it is possible to perform real-time monitoring of hybridization between target single stranded PCR products obtained by using as substrates DNA isolated from normal or heterozygous subjects, and homozygous W1282X CF samples and oligonucleotide probes, therefore enabling a one-step, non-radioactive protocol to perform diagnosis. PMID- 11438997 TI - Identification of three novel 6-pyruvoyl-tetrahydropterin synthase gene mutations (226C>T, IVS3+1G>A, 116-119delTGTT) in Chinese hyperphenylalaninemia caused by tetrahydrobiopterin synthesis deficiency. AB - The enzyme 6-Pyruvoyl-tetrahydropterin synthase (PTS) deficiency is the major cause of BH(4)-deficient HPA. The frequency of BH(4)-deficient HPA was estimated to be around 30% among Chinese HPA population in Taiwan, which is much higher than that in Caucasian population (1.5-2% of HPA). Approximately 86% of Chinese BH(4)-deficient HPA was found to be caused by PTS-deficiency. Seven mutations - namely R25G, N52S, V56M, V70D, P87S, D96N, and T106M - had been identified in Chinese PTS-deficient patients previously. In this study, five additional mutations in the PTS gene, namely 200C>T (T67M), 226C>T (L76F), IVS3+1G>A (K54X), 116-119delTGTT (K38X) and 169-171delGTG (V57del), were identified by PCR and DNA sequencing in Chinese PTS-deficient patients. The 116-119delTGTT introduces a frameshift stop after lysine of codon 38 (K38X). The G-to-A transition at the consensus sequence of splicing donor site of exon 3 (IVS3+1G>A) resulted in exon 3 skipping of the PTS transcript and caused a frameshift stop after lysine of codon 54 (K54X). The T67M and V57del mutations have been found in Caucasian PTS deficient patients, while the L76F, IVS3+1G>A, and K38X mutations are novel. None of 100 normal alleles screened was found to have the L76F substitution, which indicated that the L76F substitution is a mutation causing PTS deficiency. Hum Mutat 18:83, 2001. PMID- 11438998 TI - Twelve novel mutations in the tissue-nonspecific alkaline phosphatase gene (ALPL) in patients with various forms of hypophosphatasia. AB - Hypophosphatasia is a rare inherited disorder characterized by defective bone mineralization and deficiency of serum and tissue liver/bone/kidney tissue alkaline phosphatase (L/B/K ALP) activity. We report here the characterization of tissue-nonspecific alkaline phosphatase (TNSALP) gene mutations in a series of 11 families affected by various forms of hypophosphatasia. Nineteen distinct mutations were found, 7 of which were previously reported. Eleven of the 12 new mutations were missense mutations (Y11C, A34V, R54H, R135H, N194D, G203V, E218G, D277Y, F310G, A382S, V406A), the last one (998-1G>T) was a mutation affecting acceptor splice site. PMID- 11438999 TI - Molecular analysis of Bruton's tyrosine kinase gene in Spain. AB - Mutations in Bruton's tyrosine kinase (BTK) gene result in X linked agammaglobulinemia (XLA). Using Single Strand Conformation Polymorphism (SSCP) followed by direct sequencing 21 mutations were found in 27 patients with an XLA phenotype from 21 unrelated families. We identified 13 novel and 8 known mutations: seven missense (R288W, R544G, P566S, K430E; K374N, L512P, R544S), 5 nonsense (Q196X, Y361X, L249X, Q612X, Q466X), 2 deletions of one nucleotide (A207fsX216, Q612fsX648), 2 deletion-insertions (V219fsX227, K218fsX228), one insertion of two nucleotides (S572fsX587) and 4 point mutations in donor/acceptor splice sites (g.IVS1+1G>C, g.IVS6+5G>A, g.IVS10+1G>T, g.IVS13-1GG>CT). Carrier detection was performed in 18 mothers. Only in one case the mutation was found to be de novo. Additionally, BTK mutations were not found in four patients without family history, but with XLA-compatible phenotype. Hum Mutat 18:84, 2001. PMID- 11439000 TI - Pattern of connexin 26 (GJB2) mutations causing sensorineural hearing impairment in Ghana. AB - Mutations of the connexin 26 gene (GJB2) were studied in 365 apparently unrelated individuals with profound nonsyndromic, sensorineural hearing impairment from Ghana, West Africa. Among 121 mutated chromosomes found, 110 carried the previously described R143W mutation. A total of 6 novel mutations: L79P, V178A, R184Q, A197S, I203K, and L214P, were identified, whereby I203K was based on a dinucleotide exchange and R184Q appeared to be dominant. The GJB2 variants found in Ghana tend to comprise less nonsense and frameshift mutations and more mutations located in the C-terminal half of the molecule than the variants found in other parts of the world. PMID- 11439001 TI - Pseudoxanthoma elasticum: Point mutations in the ABCC6 gene and a large deletion including also ABCC1 and MYH11. AB - Pseudoxanthoma elasticum (PXE) is a mendelian disorder characterized by calcification of elastic fibers in skin, arteries, and retina. It results in dermal lesions, arterial insufficiency and retinal hemorrhages, leading to macular degeneration. PXE is transmitted either as an autosomal dominant or recessive trait and several sporadic cases have been observed. Mutations in the ABCC6 gene have been identified very recently in patients. Here, we report on a large Italian family affected by pseudoxanthoma elasticum for which linkage analysis had pointed to a region encompassing markers D16S3069-D16S405-D16S3103; hemizygosity of marker D16S405 allowed us to detect a submicroscopic deletion of at least 900 kb involving ABCC6, ABCC1, and MYH11. Mutation analysis on the other allele of the family, as well as on two additional sporadic cases, revealed nonsense (Y227X, R518X, R1164X) and frame-shift (c.960delC) mutations in ABCC6 (MRP6) further confirming the role of this multi-drug resistance gene in the etiology of pseudoxanthoma elasticum. Furthermore, clinical re-examination of members of the family harboring the deletion led to the detection of additional features, potentially caused by the deletion of the MYH11 gene. In the course of the analysis five nonpathogenic variants were found in ABCC6: 1233T>C, 1245G>A, 1838 T>G (V614A), 1890C>G, and 3506+83C>A. Hum Mutat 18:85, 2001. PMID- 11439002 TI - Surveillance of antibiotic resistance in European ICUs. AB - Antibiotic resistance among bacteria causing hospital-acquired infections poses a threat, particularly to patients in intensive care units (ICUs). In order to control the spread of resistant bacteria, local, regional and national resistance surveillance data must be used to develop efficient intervention strategies. In an attempt to identify national differences and the dynamics of antibiotic resistance in European ICUs, data have been merged from several networks of resistance surveillance performed during the 1990s. It should be stressed, however, that comparisons of results from different studies using different methods and different population samples must be made with caution. Antibiotic resistance across all species and drugs was, with some exceptions, highest in southern European countries and Russia, and lowest in Scandinavia. More effective strategies are needed to control the selection and spread of resistant organisms. Antibiotic intervention policies, efficient infection control measures and an overall awareness of the serious implications at public health level will contribute to the management of antibiotic resistance. PMID- 11439003 TI - Hospital infection control in Malaysia. PMID- 11439004 TI - Autoclave performance and operator knowledge of autoclave use in primary care: a survey of UK practices. AB - Changes in the delivery of health care have increased the demand for minor surgical and screening procedures in general practice. This has increased the risk of cross-infection with blood-borne viruses and the demand for sterile instruments. The aim of this study was to investigate the knowledge and training of medical personnel in England and Wales on aspects of autoclave use, and to test the effectiveness of their practice autoclaves. An anonymous postal questionnaire and autoclave performance survey using biological indicators was made of 700 general medical practitioners selected at random from FHSA lists from 12 Health Authorities in England and Wales. The overall response rate was 53.1% (N= 372) comprising 10% general practitioners and 90% practice nurses. Eighty-two percent of respondents used autoclaves with a mean age of 2.5 years, of which 91% had been serviced in the past year. While 35% of respondents made daily observation of gauges, 19% did not routinely monitor autoclave effectiveness. Six autoclaves failed to sterilize the spore test ampoules. Fourteen percent of respondents did not autoclave instruments after every patient. Only 33.1% reported wearing gloves during minor operations. Fifty five percent had training in cross-infection prevention. It was concluded that although there has been improvement in instrument decontamination procedures in general practice compared with previous surveys, further education of medical practitioners and practice nurses in the use of autoclaves and infection prevention and control is indicated. The failure of 2% of the autoclaves to kill spores suggests the need for increased monitoring of autoclave performance. Two-thirds of practitioners exposed themselves to increased risk of infection by carrying out minor surgery without the protection of gloves. PMID- 11439005 TI - The use of palivizumab monoclonal antibody to control an outbreak of respiratory syncytial virus infection in a special care baby unit. AB - An outbreak of respiratory syncytial virus (RSV) infection affected seven premature infants in a special care baby unit during November and December 1999. Conventional infection control measures (cohorting infected babies, strict reinforcement of the use of gloves and aprons, emphasis on hand disinfection) failed to prevent spread. Palivizumab, a respiratory syncytial virus monoclonal antibody, was given to eight high-risk preterm infants. There were no further cases of RSV in the unit and none of the babies given palivizumab developed RSV. One baby who acquired RSV during the outbreak (and who was not given palivizumab) was subsequently admitted to hospital with another episode of RSV bronchiolitis. The role of palivizumab in the control of hospital outbreaks of RSV infection merits further investigation. PMID- 11439006 TI - Low rate of Candida parapsilosis-related colonization and infection in hospitalized preterm infants: a one-year prospective study. AB - We determined the rate of Candida parapsilosis colonization in preterm neonates (NN) and the relationship between colonization and systemic infection through a prospective study in the Neonatal Intensive Care Unit of a university hospital. All NN born at a gestational age of 32 weeks or less were included. Specimens from rectum, mouth and retro-auricular skin were obtained at admission and weekly thereafter. All samples were inoculated on to Sabouraud agar, CHROMagar and Dixon media. Candida species were identified using API Candida and API 20C. DNA analysis was performed using pulse field gel electrophoresis.Fifty-four patients were included (mean age: 30 +/- 1.5 weeks; mean birthweight: 1347 +/- 301 g; male: 40%). Fungal colonization was detected in 43 (79.6%). Causative agents were C. parapsilosis (N= 7);Malassezia furfur (N= 30);C. albicans (N= 21), C. guillermondii (N= 1). No sample was positive for two different yeasts at the same time. C. parapsilosis colonization included anal (N= 6), buccal (N= 1), and skin (N= 2). The average age at time of colonization was 17.8+/-9.8 days. Neither fungal septicaemia nor death were observed in colonized infants. Two central venous catheters were found to be colonized, one with C. parapsilosis and one with M. furfur. Logistic regression showed a link between colonization and gestational age alone. Three different DNA profiles were observed. This study suggests that in our units, the occurrence of C. parapsilosis colonization is low and bears no relation to systemic infection. The systematic identification of C. parapsilosis carriers for the purposes of isolation and preventive treatment does not appear to be warranted. PMID- 11439007 TI - Relationship between environmental fungal contamination and the incidence of invasive aspergillosis in haematology patients. AB - Invasive aspergillosis (IA) is a major opportunistic infection in haematology patients. Spore inhalation is the usual route of Aspergillus infection, suggesting a determining role of environmental contamination by spores in the epidemiology of IA. We prospectively examined the relationship between environmental contamination by Aspergillus and other fungal species and the incidence of invasive nosocomial aspergillosis (INA) in a bone marrow transplantation unit and two haematology wards. During a four-year period, levels of air and surface fungal contamination were determined bi-monthly in patients' rooms (some equipped with HEPA filters and LAF systems), and various common sites in each ward (corridors, nursing stations, etc.). Results were compared to the incidence of INA. A total of 3100 air and 9800 surface samples were collected, and 79 cases of IA were diagnosed, of which 64 were probably or possibly INA. Patterns of fungal contamination were comparable in the three wards, with a gradient ranging from high levels in common sites to a virtual absence in rooms equipped with HEPA filters and LAF systems. Using a regression model, a significant relationship was found between the incidence of INA and the degree of fungal contamination of air and surfaces in conventional patient rooms (not equipped with HEPA) and common sites. This study shows that in a non-epidemic setting, there is a significant relationship between environmental fungal contamination in haematology wards and the incidence of INA. Our findings underline the importance of environmental surveillance and strict application of preventive measures. PMID- 11439008 TI - Asymptomatic carriage of Klebsiella pneumoniae producing extended-spectrum beta lactamase by patients in a neurological early rehabilitation unit: management of an outbreak. AB - During 11 months 58 extended-spectrum beta-lactamase-producing Klebsiella pneumoniae (ESBL-Kp) isolates were grown from 10 patients on a neurological early rehabilitation unit. The patients had no signs of infection but were colonized in the nose and trachea, and unusually only one had colonization in the gut. A single clone of ESBL-Kp was identified by pulse field gel electrophoresis. Strong hygienic precautions similar to those for Methicillin-resistant Staphylococcus aureus patients prevented spread of the bacteria to other wards. However, rehabilitation for patients with severe neurological failures made it very difficult to follow hygienic requirements. Disinfection of mucous membranes was difficult. Eventually the application of a nasal spray containing povidone-iodine proved to be successful. PMID- 11439009 TI - Comparison of the mycobactericidal activity of ortho- phthalaldehyde, glutaraldehyde and other dialdehydes by a quantitative suspension test. AB - The mycobactericidal activity of various dialdehydes has been assessed by a quantitative suspension test in both 'clean' and 'dirty' conditions. Test organisms consisted of glutaraldehyde (GTA)-sensitive strains of Mycobacterium chelonae NCTC 946, M. abscessus NCTC 10882, two GTA-resistant M. chelonae strains and M. terrae NCTC 10856 (a proposed M. tuberculosis surrogate). The aldehydes tested were a new high-level disinfectant, ortho-phthalaldehyde (OPA) at 0.5% (v/v) unadjusted pH 6.5 and pH 8, GTA at 0.5% (v/v) pH 8, glyoxal at 0.5% (v/v) pH 8 and 10% (v/v) unadjusted pH 2.8, malonaldehyde sodium salt (NaMDA) at 0.5% (w/v) pH 8 and 10% (w/v) unadjusted pH 7.5 and succinaldehyde at 0.5% (v/v) pH 8. Results showed that 0.5% acidic and alkaline OPA were rapidly mycobactericidal, under both 'clean' and 'dirty' conditions, and more importantly were active against GTA-resistant strains. The washer disinfector isolates of M. chelonae were, as expected, extremely resistant to 0.5% GTA which was slowly mycobactericidal against the other strains. Glyoxal, NaMDA and succinaldehyde were ineffective against all the strains investigated. However, a high concentration of glyoxal exhibited a slow mycobactericidal activity except with M. terrae NCTC 10856, but this was not observed with NaMDA. This evaluation, using a quantitative suspension test based on a European standard, supported the claim that OPA is an effective choice as a high-level disinfectant for medical devices. PMID- 11439010 TI - Evaluation of a patient-empowering hand hygiene programme in the UK. AB - Partners in Your Care, a patient education behavioral model for increasing handwashing compliance and empowering the patient with responsibility for their care was evaluated in an acute care hospital in Oxford, UK. A controlled prospective intervention study comparing medical and surgical patients was performed. Ninety-eight patients were eligible for the study. Thirty-nine patients (40%) agreed to participate in the programme Partners in Your Care by asking all healthcare workers who were going to have direct contact with them "Did you wash your hands?" Compliance with the programme was measured through soap/alcohol usage and handwashings per bed day before and after its introduction. Partners in Your Care increased handwashing on average 50%. Healthcare workers washed hands more often with surgical patients than with medical (P< 0.05). Alcohol gel was used on less than 1% of occasions. Sixty-two percent of patients in study felt at ease when asking healthcare workers "Did you wash your hands?" Seventy-eight percent received a positive response (washed hands). All patients asked nurses, but only 35% asked physicians. Partners in Your Care increased handwashing compliance in the UK. This programme empowers patients with responsibility for their care, provides infection control staff with a continuing means for providing handwashing education without additional staff and can save costs for a hospital. PMID- 11439011 TI - Investigation of an outbreak of multi-drug resistant Acinetobacter baumannii in an intensive care burns unit. AB - Over a three month period there was an outbreak of infection, due to a multi-drug resistant Acinetobacter baumannii in the intensive care burns unit with spread of infection to other patients, both within the unit and elsewhere in the hospital. Microbiological sampling of the environment and of the healthcare workers' (HCWs) hands were carried out. Strain relatedness of the isolates was confirmed by pulsed field gel electrophoresis. Fifteen patients were involved in the outbreak, whose infections were all hospital-acquired. The burns room environment was contaminated with the A. baumannii, as was the door handle of the door leading from the ante-chamber between both rooms. This allowed the hands of HCWs to be contaminated by A. baumannii despite appropriate handwashing procedures prior to leaving the rooms. Two staff members were colonized with A. baumannii. One HCW who was directly involved in patient care was found to be "heavily" colonized, the other, with less patient contact, was only "lightly" colonized. Review of handwashing practices revealed that chlorhexidine/alcohol hand wash solution was not used by the HCW whose hands were heavily colonized.A combination of a review of handwashing practice, education about the spread of bacteria via hands and contaminated environment, and the revision of infection control procedures in the unit contributed to a prompt termination of the outbreak. PMID- 11439012 TI - "Second-look" cytotoxicity: an evaluation of culture plus cytotoxin assay of Clostridium difficile isolates in the laboratory diagnosis of CDAD. AB - Clostridium difficile is one of the most frequent causes of hospital-acquired diarrhoea. Our objective was to prove that some stool samples with a direct negative cytotoxicity assay may indeed harbour toxigenic C. difficile and that this can be demonstrated by performing a "second-look" cytotoxicity assay using the isolated C. difficile strains. Over an eight-year period (1992-1999), the 8241 stool samples submitted for direct cell culture from patients with suspected C. difficile-associated diarrhoea (CDAD) were simultaneously plated on cycloserine cefoxitin fructose agar. C. difficile strains isolated from samples with a negative direct cell culture assay were re-tested for toxin production "second-look" cell culture assay). Using both methods 6423 samples (78%) were negative. Of the remaining 1818 samples, 127 (7%) yielded C. difficile isolates which were confirmed as non-producers of toxin by both methods, 1437 (85%) were positive in direct cell culture assay, and 254 were positive only after the "second-look" cell culture assay. Thus, our approach allowed us to detect an extra 15% of toxin-producing strains that could have gone undetected otherwise.The combination of direct-cell culture assay, culture for toxigenic C. difficile and "second-look" cell culture assay enhances the potential for diagnosis of CDAD and enables us to be more efficient with our patient care resources. PMID- 11439013 TI - Bacterial contamination of uniforms. AB - Microbiological sampling of nurses' uniforms was undertaken using a Casella slit sampler. Staphylococcus aureus, Clostridium difficile and vancomycin-resistant enterococci were detected on uniforms both before and after a span of duty. Recommendations for provision and changing of nurses' uniforms are made. PMID- 11439014 TI - "Road signs" approach to hand hygiene. PMID- 11439015 TI - Monitoring glutaraldehyde dilution in automated washer/disinfectors. PMID- 11439016 TI - Handwashing: answering questions and pursuing compliance. PMID- 11439017 TI - A phylogenetic analysis elucidating a case of patient-to-patient transmission of hepatitis C virus during surgery. PMID- 11439018 TI - DNA binding and cleavage selectivity of the Escherichia coli DNA G:T-mismatch endonuclease (vsr protein). AB - The Escherichia coli vsr endonuclease recognises T:G base-pair mismatches in double-stranded DNA and initiates a repair pathway by hydrolysing the phosphate group 5' to the incorrectly paired T. The gene encoding the vsr endonuclease is next to the gene specifying the E. coli dcm DNA-methyltransferase; an enzyme that adds CH3 groups to the first dC within its target sequence CC[A/T]GG, giving C5MeC[A/T]GG. Deamination of the d5MeC results in CT[A/T]GG in which the first T is mis-paired with dG and it is believed that the endonuclease preferentially recognises T:G mismatches within the dcm recognition site. Here, the preference of the vsr endonuclease for bases surrounding the T:G mismatch has been evaluated. Determination of specificity constant (kst/KD; kst = rate constant for single turnover, KD = equilibrium dissociation constant) confirms vsr's preference for a T:G mismatch within a dcm sequence i.e. CT[A/T]GG (the underlined T being mis-paired with dG) is the best substrate. However, the enzyme is capable of binding and hydrolysing sequences that differ from the dcm target site by a single base-pair (dcm star sites). Individual alteration of any of the four bases surrounding the mismatched T gives a substrate, albeit with reduced binding affinity and slowed turnover rates. The vsr endonuclease has a much lower selectivity for the dcm sequence than type II restriction endonucleases have for their target sites. The results are discussed in the light of the known crystal structure of the vsr protein and its possible physiological role. PMID- 11439019 TI - Interrupting the template strand of the T7 promoter facilitates translocation of the DNA during initiation, reducing transcript slippage and the release of abortive products. AB - We have explored the effects of a variety of structural and sequence changes in the initiation region of the phage T7 promoter on promoter function. At promoters in which the template strand (T strand) is intact, initiation is directed a minimal distance of 5 nt downstream from the binding region. Although the sequence of the DNA surrounding the start site is not critical for correct initiation, it is important for melting of the promoter and stabilization of the initiation complex. At promoters in which the integrity of T strand is interrupted by nicks or gaps between -5 and -2 the enzyme continues to initiate predominately at +1. However, under these conditions there is a decrease in the release of abortive products of 8-10 nt, a decrease in the synthesis of poly(G) products (which arise by slippage of the nascent transcript), and a defect in displacement of the RNA. We propose that unlinking the binding and initiation regions of the T strand changes the manner in which this strand is retained in the abortive complex, reducing or eliminating the need to pack or "scrunch" the strand into the complex during initiation and lowering a thermodynamic barrier to its translocation. PMID- 11439020 TI - Long-range identification of hepatocyte nuclear factor-3 (FoxA) high and low affinity binding sites with a chimeric nuclease. AB - Identifying the complete set of transcription factors that bind the promoter and other regulatory regions of a gene of interest is an essential step in functional genomics. We have developed an original assay for the systematic detection of hepatocyte nuclear factor-3 (HNF-3) binding sites within cloned promoters. This assay is based on expression of a recombinant enzyme, HNF-3beta/FN, that is comprised of the rat HNF-3beta DNA-binding domain and the non-specific nuclease domain of the FokI restriction enzyme. Southern analysis of target plasmids with proven HNF-3 binding sites showed that HNF-3beta/FN was able to specifically cut both DNA strands in the vicinity of these binding sites, whereas mutagenized binding sites were no longer cleaved. Likewise, as yet undescribed HNF-3 binding sites were detected easily over a distance spanning several thousand bases. The functionality of such binding sites was confirmed by electromobility shift assay. Furthermore, the extent of cleavage by HNF-3beta/FN at a given binding site was tightly correlated with the affinity of a natural HNF-3beta molecule for this site. This novel approach can be extended to other transcription factors for long range identification of functional transcription factor binding sites in genes. PMID- 11439021 TI - Construction of an Escherichia coli knockout strain for functional analysis of tRNA(Asp). AB - The specific aminoacylation of tRNA is critical for translation of the genetic code. A molecular description of aminoacylation requires knowledge of the relevant three-dimensional structures, biochemical parameters and the structure function relationship of the synthetase and its substrate tRNA. Extensive structural and biochemical data are available on the aspartic acid system of Escherichia coli, but there is a paucity of cellular functional data. We have developed a system to overcome this deficiency by engineering an E. coli knockout tRNA(Asp) strain, thereby allowing a penetrating analysis of tRNA(Asp) structure and function under conditions that prevail in the cell. PMID- 11439022 TI - Plasmid systems to study RNA function in Escherichia coli. AB - Determining the functional activity of an essential RNA in vivo presents special challenges. We have devised an in vivo analysis of alternative forms of an essential tRNA gene in Escherichia coli knockout cells using either a plasmid switch or a regulated two-plasmid system. The model system is presented together with a description of the new plasmids and procedures necessary to effect these analyses. The system is readily adaptable to non-essential RNAs. PMID- 11439023 TI - Mitochondrial DNA from the liverwort Marchantia polymorpha: circularly permuted linear molecules, head-to-tail concatemers, and a 5' protein. AB - Mapping predicts that the mitochondrial genome of the liverwort Marchantia polymorpha exists as a circular molecule, although nearly all the mitochondrial DNA (mtDNA) is found as genome-sized and multigenomic molecules in linear and branched form. We used restriction enzymes with one recognition site per genome, end-specific exonucleases and pulsed-field gel electrophoresis (PFGE) to analyze the arrangement of genomic units and the terminal structure of the molecules. We find a head-to-tail arrangement in the concatemers and circular permutation in both the monomeric and multigenomic molecules. The termini contain covalently bound protein at the 5' end and an open (unblocked) 3' end. We find that the standard in-gel procedure used to prepare large DNA molecules for PFGE may introduce extraction artifacts leading to erroneous conclusions about the termini. These artifacts can be reduced by omitting high salt (high EDTA) and protease during mitochondrial lysis. Our results suggest that the mtDNA may use a T4 phage-like mechanism of replication and that the linear molecules may be due to strand breaks mediated by type II topoisomerase. PMID- 11439024 TI - Analysis of the NF-kappaB p50 dimer interface by diversity screening. AB - An in vivo screen has been devised for NF-kappaB p50 activity in Escherichia coli exploiting the ability of the mammalian transcription factor to emulate a prokaryotic repressor. Active intracellular p50 was shown to repress the expression of a green fluorescent protein reporter gene allowing for visual screening of colonies expressing active p50 on agar plates. A library of mutants was constructed in which the residues Y267, L269, A308 and V310 of the dimer interface were simultaneously randomised and twenty-five novel functional interfaces were selected which repressed the reporter gene to similar levels as the wild-type protein. The leucine-269 alanine-308 core was repeatedly, but not exclusively, selected from the library whilst a diversity of predominantly non polar residues were selected at positions 267 and 310. These results indicate that L269 and A308 may form a hot spot of interaction and allow an insight into the processes of dimer selectivity and evolution within this family of transcription factors. PMID- 11439025 TI - Genetic analysis of an archaeal Holliday junction resolvase in Escherichia coli. AB - The study of genes and proteins in heterologous model systems provides a powerful approach to the analysis of common processes in biology. Here, we show how the bacterium Escherichia coli can be exploited to analyse genetically and biochemically the activity and function of a Holliday junction resolving enzyme from an archaeal species. We have purified and characterised a member of the newly discovered Holliday junction cleaving (Hjc) family of resolvases from the moderately thermophilic archaeon Methanobacterium thermoautotrophicum and demonstrate that it promotes DNA repair in resolvase-deficient ruv mutants of E. coli. The data presented provide the first direct evidence that such archaeal enzymes can promote DNA repair in vivo, and support the view that formation and resolution of Holliday junctions are key to the interplay between DNA replication, recombination and repair in all organisms. We also show that Hjc promotes DNA repair in E. coli in a manner that requires the presence of the RecG branch migration protein. These results support models in which RecG acts at a replication fork stalled at a lesion in the DNA, catalysing fork regression and forming a Holliday junction that can then be acted upon by Hjc. PMID- 11439026 TI - Building novel binding ligands to B7.1 and B7.2 based on human antibody single variable light chain domains. AB - Ligands specific for B7.1 (CD80) and B7.2 (CD86) have applications in disease indications that require inhibition of T-cell activity. As we observed significant sequence and structural similarity between the B7-binding ligand, cytotoxic T-lymphocyte associated protein-4 (CTLA-4), and antibody variable light chain domains (VLs), we have explored the possibilities of making novel B7 binding molecules based on single VL domains. We first describe the "rational" design and construction of a VL/CTLA-4 hybrid molecule in which we have grafted both the CDR1 and CDR3-like loops of CTLA-4 onto a single VL light chain, at sites determined by sequence and structure-based alignment. This molecule was secreted as a soluble product from Escherichia coli, but did not show any binding to B7.1 and B7.2. In a second approach we constructed a VL library in which human VL genes derived from B-cells were spiked with the CDR3-like loop of CTLA-4 and further diversified by DNA shuffling. This library was displayed on phage, and after selection gave B7.1 binding ligands which competed with CTLA-4. In order to evaluate the possible general utility of VL domains as binding ligands, we have constructed a non-biased VL library. From this DNA-shuffled human VL library we have selected single VL domains specific for B7.1, B7.2 or human IgG. Two B7.1 specific VL ligands and one B7.2-specific VL ligand showed competition with CTLA 4. One candidate VL domain-specific for B7.1 was affinity matured by simultaneous randomisation of all CDR loops using DNA shuffling with degenerate CDR-spiking oligonucleotides. From this library, a single VL domain with affinity of 191 nM for B7.1 was obtained, which also showed binding to B7.1 in situ. This VL had mutations in CDR1 and CDR3, indicating that antigen recognition for this single VL is most likely mediated by the same regions as in the VL domain of whole antibodies. The B7.1 and B7.2-specific VL domains described in this study may form the basis of a new family of immunomodulatory recombinant molecules. Furthermore, our studies suggest that it is feasible to create specific single VL domains to diverse targets as is the case for single VH domains. PMID- 11439027 TI - A semi-synthetic repertoire of intrinsically stable antibody fragments derived from a single-framework scaffold. AB - We report the design, construction and use of an antibody bacteriophage display library built on the scaffold of a single-chain variable fragment (scFv) previously proven to be functionally expressed in the reducing environment of both bacterial and plant cytoplasm and endowed with intrinsic high thermodynamic stability. Four amino acid residues of the third hypervariable loop (CDR3) of both VH and VL were combinatorially mutated, generating a repertoire of approximately 5x10(7) independent scFvs, cloned in a phagemid vector. The ability of the antibody phage library to yield specific binders was tested by biopanning against several antigens. Successful selection of fully active scFvs was obtained, confirming the notion that combinatorial mutagenesis of few amino acid residues centrally located in the antigen-binding site is sufficient to provide binding specificities against virtually any target. High yields of both soluble and phage antibodies were obtained in Escherichia coli. Maintenance of the cognate scFv antibody stability in the newly selected scFv fragments was demonstrated by guanidinium chloride denaturation/renaturation studies and by soluble antibody expression in the bacterial cytoplasm. The antibody library described here allows the isolation of new stable binding specificities, potentially exploitable as immunochemical reagents for intracellular applications. PMID- 11439028 TI - Crystal structures of YBHB and YBCL from Escherichia coli, two bacterial homologues to a Raf kinase inhibitor protein. AB - In rat and human cells, RKIP (previously known as PEBP) was characterized as an inhibitor of the MEK phosphorylation by Raf-1. In Escherichia coli, the genes ybhb and ybcl possibly encode two RKIP homologues while in the genomes of other bacteria and archaebacteria other homologous genes of RKIP have been found. The parallel between the cellular signaling mechanisms in eukaryotes and prokaryotes suggests that these bacterial proteins could be involved in the regulation of protein phosphorylation by kinases as well. We first showed that the proteins YBHB and YBCL were present in the cytoplasm and periplasm of E. coli, respectively, after which we determined their crystallographic structures. These structures verify that YBHB and YBCL belong to the same structural family as mammalian RKIP/PEBP proteins. The general fold and the anion binding site of these proteins are extremely well conserved between mammals and bacteria and suggest functional similarities. However, the bacterial proteins also exhibit some specific structural features, like a substrate binding pocket formed by the dimerization interface and the absence of cis peptide bonds. This structural variety should correspond to the recognition of multiple cellular partners. PMID- 11439029 TI - The dimerization domain of HNF-1alpha: structure and plasticity of an intertwined four-helix bundle with application to diabetes mellitus. AB - Maturity-onset diabetes mellitus of the young (MODY) is a human genetic syndrome most commonly due to mutations in hepatocyte nuclear factor-1alpha (HNF-1alpha). Here, we describe the crystal structure of the HNF-1alpha dimerization domain at 1.7 A resolution and assess its structural plasticity. The crystal's low solvent content (23%, v/v) leads to tight packing of peptides in the lattice. Two independent dimers, similar in structure, are formed in the unit cell by a 2-fold crystallographic symmetry axis. The dimers define a novel intertwined four-helix bundle (4HB). Each protomer contains two alpha-helices separated by a sharp non canonical turn. Dimer-related alpha-helices form anti-parallel coiled-coils, including an N-terminal "mini-zipper" complementary in structure, symmetry and surface characteristics to transcriptional coactivator dimerization cofactor of HNF-1 (DCoH). A confluence of ten leucine side-chains (five per protomer) forms a hydrophobic core. Isotope-assisted NMR studies demonstrate that a similar intertwined dimer exists in solution. Comparison of structures obtained in multiple independent crystal forms indicates that the mini-zipper is a stable structural element, whereas the C-terminal alpha-helix can adopt a broad range of orientations. Segmental alignment of the mini-zipper (mean pairwise root-mean square difference (rmsd) in C(alpha) coordinates of 0.29 A) is associated with a 2.1 A mean C(alpha) rmsd displacement of the C-terminal coiled-coil. The greatest C-terminal structural variation (4.1 A C(alpha) rmsd displacement) is observed in the DCoH-bound peptide. Diabetes-associated mutations perturb distinct structural features of the HNF-1alpha domain. One mutation (L12H) destabilizes the domain but preserves structural specificity. Adjoining H12 side-chains in a native-like dimer are predicted to alter the functional surface of the mini-zipper involved in DCoH recognition. The other mutation (G20R), by contrast, leads to a dimeric molten globule, as indicated by its 1H-NMR features and fluorescent binding of 1 anilino-8-naphthalene sulfonate. We propose that a glycine-specific turn configuration enables specific interactions between the mini-zipper and the C terminal coiled-coil. PMID- 11439030 TI - Determination of the pK(a) of the four Zn2+-coordinating residues of the distal finger motif of the HIV-1 nucleocapsid protein: consequences on the binding of Zn2+. AB - The nucleocapsid protein NCp7 of human immunodeficiency virus type 1 is characterized by two highly conserved CCHC motifs that bind Zn2+ strongly. To elucidate the striking pH-dependence of the apparent Zn2+-binding constants of these motifs further, we investigated, using 1H NMR, potentiometry and fluorescence spectroscopy, the acid-base properties of the four Zn2+-coordinating residues of (35-50)NCp7, a peptide corresponding to the distal finger motif of NCp7. With the exception of the H(beta2) proton of Cys39, the pH-dependence of the H(beta) proton resonances of the three Cys residues and, the H(delta) and H(epsilon) resonances of His44 in the apopeptide could be fitted adequately with a single pK(a). This suggests that the protonating groups are non-interacting, a feature that was confirmed by a potentiometric titration. The pK(a) of His44, Cys36, Cys39, and Cys49 in the apopeptide were found to be 6.4, 8.0, 8.8 and 9.3, respectively. Accordingly, the deprotonation is almost sequential and may thus induce a sequential binding of Zn2+ to the four coordinating residues. The high pK(a) of Cys49 is probably related to the negative charge of the neighboring Asp48. Such a high pK(a) may be a general feature in nucleocapsid proteins (NCs), since an acidic residue generally occupies the (i-1) position of the C-terminal Cys residue of single-finger NCs and distal finger motifs in two-finger NCs. Molecular dynamics simulation suggested the formation of a hydrogen bonded network that weakly structured the Cys36-Cys39 segment in the apopeptide. This network depends on the protonation state of Cys36 and may thus explain the biphasic behavior of the pH-dependence of the Cys39 H(beta2) resonance. Finally, the pK(a) values were used to build up a model describing the coordination of Zn2+ to (35-50)NCp7 at equilibrium. It appears that each protonation step of the coordination complex decreases the Zn2+-binding constant by about four orders of magnitude and that a significant dissociation of Zn2+ from the holopeptide can be achieved in acidic cell compartments. PMID- 11439031 TI - Characterization of the folding kinetics of a three-helix bundle protein via a minimalist Langevin model. AB - We use a simple off-lattice Langevin model of protein folding to characterize the folding and unfolding of a fast-folding, 46 residue three-helix bundle. Under conditions at which the C-terminal helix is 30 % stable, we observe a clear three state folding mechanism. In the on-pathway intermediate state, the middle and C terminal helices are folded and in contact with each other, while the N-terminal region remains disordered. Nevertheless, under these conditions this intermediate is thermodynamically unstable relative to its unfolded state. The first and highest folding barrier corresponds to the organization of the hinge between the middle and C-terminal helices. A subsequent major barrier corresponds to the organization of the hinge between the middle and N-terminal helices. Hyperstabilizing the hinge regions leads to twice the folding rate that is obtained from hyperstabilizing the helices, even though much fewer contacts are involved in hinge hyperstabilization than in helix hyperstabilization. Unfolding follows single-exponential kinetics, even at temperatures only slightly above the folding transition temperature. PMID- 11439032 TI - Assessing the balance between protein-protein interactions and enzyme-substrate interactions in the channeling of intermediates between polyketide synthase modules. AB - 6-Deoxyerythronolide B synthase (DEBS) is the modular polyketide synthase (PKS) that catalyzes the biosynthesis of 6-deoxyerythronolide B (6-dEB), the aglycon precursor of the antibiotic erythromycin. The biosynthesis of 6-dEB exemplifies the extraordinary substrate- and stereo-selectivity of this family of multifunctional enzymes. Paradoxically, DEBS has been shown to be an attractive scaffold for combinatorial biosynthesis, indicating that its constituent modules are also very tolerant of unnatural substrates. By interrogating individual modules of DEBS with a panel of diketides activated as N-acetylcysteamine (NAC) thioesters, it was recently shown that individual modules have a marked ability to discriminate among certain diastereomeric diketides. However, since free NAC thioesters were used as substrates in these studies, the modules were primed by a diffusive process, which precluded involvement of the covalent, substrate channeling mechanism by which enzyme-bound intermediates are directly transferred from one module to the next in a multimodular PKS. Recent evidence pointing to a pivotal role for protein-protein interactions in the substrate-channeling mechanism has prompted us to develop novel assays to reassess the steady-state kinetic parameters of individual DEBS modules when primed in a more "natural" channeling mode by the same panel of diketide substrates used earlier. Here we describe these assays and use them to quantify the kinetic benefit of linker mediated substrate channeling in a modular PKS. This benefit can be substantial, especially for intrinsically poor substrates. Examples are presented where the k(cat) of a module for a given diketide substrate increases >100-fold when the substrate is presented to the module in a channeling mode as opposed to a diffusive mode. However, the substrate specificity profiles for individual modules are conserved regardless of the mode of presentation. By highlighting how substrate channeling can allow PKS modules to effectively accept and process intrinsically poor substrates, these studies provide a rational basis for examining the enormous untapped potential for combinatorial biosynthesis via module rearrangement. PMID- 11439033 TI - A role for highly conserved carboxylate, aspartate-140, in oxygen activation and heme degradation by heme oxygenase-1. AB - Heme oxygenase (HO) catalyzes the oxygen-dependent degradation of heme to biliverdinIXalpha, CO, and free iron ion via three sequential monooxygenase reactions. Although the distinct active-site structure of HO from cytochrome P450 families suggests unique distal protein machinery to activate molecular oxygen, the mechanism and the key amino acid for the oxygen activation have not been clear. To investigate the functionality of highly conserved polar amino acids in the distal helix of HO-1, we have prepared alanine mutants: T135A, R136A, D140A, and S142A, and found drastic changes in the heme degradation reactions of D140A. In this paper, we report the first evidence that D140 is involved in the oxygen activation mechanism in HO-1. The heme complexes of HO mutants examined in this study fold and bind heme normally. The pK(a) values of the iron-bound water and autoxidation rates of the oxy-form are increased with R136A, D140A, and S142A mutations, but are not changed with T135A mutation. As the wild-type, T135A, R136A, and S142A degrade heme to verdohemeIXalpha with H(2)O(2) and to biliverdinIXalpha with the NADPH reductase system. On the other hand, D140A heme complex forms compound II with H(2)O(2), and no heme degradation occurs. For the NADPH reductase system, the oxy-form of D140A heme complex is accumulated in the reaction, and only 50% of heme is degraded. The stopped flow experiments suggest that D140A cannot activate iron-bound dioxygen and hydroperoxide properly. To investigate the carboxylate functionality of D140, we further replaced D140 with glutamic acid (D140E), phenylalanine (D140F), and asparagine (D140N). D140E degrades heme normally, but D140N shows reactivity similar to that of D140A. D140F loses heme degradation activity completely. All of these results indicate that the carboxylate at position 140 is essential to activate the iron-bound dioxygen and hydroperoxide. On the basis of the present findings, we propose an oxygen activation mechanism involving the hydrogen-bonding network through the bridging water and D140 side chain. PMID- 11439034 TI - Differential rates of reversibility of ecteinascidin 743-DNA covalent adducts from different sequences lead to migration to favored bonding sites. AB - Ecteinascidin 743 (Et 743), one of a series of structurally related antitumor antibiotics isolated from a marine tunicate, is currently in phase II clinical trials. Et 743 alkylates guanine N2 through the minor groove of DNA. Hydrogen bonding networks that associate the drug with a three base pair DNA recognition site have been proposed to contribute to both the reactivity and the stability of the Et 743-DNA adduct. Here, we report that the reaction of Et 743 with DNA is reversible under nondenaturing conditions and that the rate of this reverse reaction depends critically upon the DNA-modified sequence. Quite unexpectedly, it was found that although the rates of alkylation are similar for the 5'-AGT and 5'-AGC sequences, reversal from the 5'-AGT sequence occurs faster than from the 5'-AGC sequence. Consequently, it is the differences in the rate of the reverse reaction that dictate the sequence selectivity of Et 743 toward its favored target sequence. As a direct consequence of the reversible nature of Et 743 with DNA, Et 743 can migrate from the nonfavored bonding sequence (e.g., 5'-AGT) to the favored DNA target site (e.g., 5'-AGC). The data suggest that the observed differences in the rate of reversibility arise from differences in the stability of the Et 743-DNA adduct at the 5'-AGT and 5'-AGC target sequences. On the basis of gel electrophoresis and (1)H NMR experiments, the Et 743-AGT adduct is less stable, has more dynamic motion, and produces different conformational changes in the DNA than the more stable Et 743-AGC adduct. The shuffling of Et 743-DNA adducts to the more stable alkylation sites has important implications for understanding the underlying relationship between the structural modification of DNA by Et 743 and its biological potency and efficacy in tumor cells. PMID- 11439035 TI - Selection of enantioselective acyl transfer catalysts from a pooled peptide library through a fluorescence-based activity assay: an approach to kinetic resolution of secondary alcohols of broad structural scope. AB - An assay employing a fluorescently labeled split and pool peptide library has been applied to the discovery of a new class of octapeptide catalysts for the kinetic resolution of secondary alcohols. A highly diverse library of peptide based catalysts was synthesized on solid-phase synthesis beads such that each individual bead was co-functionalized with (i) a uniform loading of a pH sensitive fluorophore and (ii) a unique peptide-based catalyst. The library was then screened for activity in acylation reactions employing (+/-)-sec phenylethanol as the substrate and acetic anhydride as the acylation agent. From the most active catalysts, a lead peptide (4) was identified that provides a selectivity-factor (k(rel)) of 8.2 upon resynthesis and evaluation under homogeneous conditions. A "directed" second-generation split and pool peptide library was synthesized such that the new peptide sequences in the library were biased toward the lead structure. Random samples of the second generation library were screened in single bead assays that revealed several new peptide-based catalysts that afford improved selectivities in kinetic resolutions. Peptide catalyst 13 proves effective for the kinetic resolution of sec-phenylethanol (k(rel) = 20), as well as eight other secondary alcohols of a broad substrate scope (k(rel) = 4 to >50). PMID- 11439036 TI - Molecular dynamics simulations of the rehydration of folded and unfolded cytochrome C ions in the vapor phase. AB - Molecular dynamics (MD) simulations have been performed to study the rehydration of compact and unfolded cytochrome c ions in the vapor phase. Experimental studies have shown that the compact conformations adsorb many more water molecules than unfolded ones when exposed to water vapor. MD simulations performed with up to 150 water molecules reproduce the key experimental observations, including a partial refolding caused by hydration. According to the calculations it is more energetically favorable to hydrate the compact conformation in the initial stages of hydration, because it is easier for a water molecule to interact simultaneously with several polar groups (due to their proximity). The protonated side chains are not favored hydration sites in the simulations because they have "self-solvation" shells which must be disrupted for the water to penetrate. For both conformations, the adsorbed water molecules are mainly located in surface crevices. PMID- 11439037 TI - Importance of planar chirality in chiral catalysts with three chiral elements: the role of planar chirality in 2'-substituted 1,1'-P,N-ferrocene ligands on the enantioselectivity in Pd-catalyzed allylic substitution. AB - A series of novel planar chiral 2'-substituted 1,1'-P,N-ferrocene ligands 9-11, 14, and 16 were prepared with diastereopurity >99:1 and found to be effective in asymmetric allylic alkylation and amination reactions. Ligand 14 furnished the highest enantiomeric excess, 98.5% and 96.5% ee in alkylation and amination reactions, respectively. The role of planar chirality in asymmetric reactions has been examined, and decisive effects on enantioselectivity as well as the control of absolute configuration in palladium-catalyzed allylic alkylation and amination reactions were observed. To clarify why and how the planar chirality governed the stereochemical outcome, X-ray crystallographic structures of eta(3)-diphenylallyl Pd complexes, (1)H NMR, (31)P NMR spectra of palladium dichloride complexes, and eta(3)-diphenylallyl Pd complexes of three 1,1'-P,N-ferrocene ligands were analyzed with the aid of COSY and 2D NOESY experiments. All results led to the conclusion that planar chirality influences the stereochemical outcome by changing or even inverting the ratio of two rotamers because of the steric interaction between a planar chiral group and the coordination site. PMID- 11439038 TI - Symmetry of N-H-N hydrogen bonds in 1,8-bis(dimethylamino)naphthalene.H+ and 2,7 dimethoxy-1,8-bis(dimethylamino)naphthalene.H+. AB - In solution, are the hydrogen bonds in monoprotonated N,N,N',N'-tetramethyl-1,8 naphthalenediamines single- or double-well? To answer this question, isotopic perturbation of equilibrium is applied to a mixture of -d(0), -d(3), -d(6), d(9), and -d(12) isotopologs. The N-methyls of the 2,7-dimethoxy analogue show intrinsic isotope shifts from the geminal CD(3) and from only one distant CD(3), an unusual stereochemical effect transmitted across the hydrogen bond. The (13)C NMR splittings and intensities at the various ring carbons of both ions are consistent with perturbation isotope shifts, intrinsic shifts, or a combination of both. The perturbation shifts mean that the protons reside in a double-minimum potential and that each ion is a pair of rapidly interconverting tautomers. The significance of this result for the role of low-barrier hydrogen bonds in enzyme catalyzed reactions is discussed. PMID- 11439039 TI - The regioselectivity of addition of organolithium reagents to enones and enals: the role of HMPA. AB - The role of polar solvents (particularly HMPA) in controlling the ratio of 1,2 to 1,4 addition of sulfur-substituted organolithium reagents to cyclohexenones and hexenal was studied. Low-temperature, multinuclear NMR studies provided quantitative information about the ratio of contact (CIP) and solvent-separated (SIP) ion pairs in solutions of dithianyllithiums and phenylthiobenzyllithiums in THF-HMPA solutions. The ratio of contact and separated ion pairs was manipulated by changes in the strength of solvation (generally through the addition of HMPA). Although the results are consistent with the CIP/SIP distribution being an important factor in determining the regioselectivity of these additions (Curtin Hammett limitations prevent a direct correlation), it cannot be the only one. Changes in diastereomeric product ratios upon addition of HMPA suggest that complexation of HMPA to lithium has two effects. First, it causes ion pair separation, which enhances 1,4 addition. Second, it lowers the Lewis acidity and catalytic effectiveness of the lithium cation, which also favors 1,4 addition. For most sulfur-stabilized lithium reagents, 2 equiv of HMPA suffice to achieve >95% 1,4 addition, whereas 4 equiv of DMPU are required to achieve identical regiochemical and stereochemical results. PMID- 11439040 TI - Functionalization of carbon nanotubes by electrochemical reduction of aryl diazonium salts: a bucky paper electrode. AB - Small-diameter (ca. 0.7 nm) single-wall carbon nanotubes are predicted to display enhanced reactivity relative to larger-diameter nanotubes due to increased curvature strain. The derivatization of these small-diameter nanotubes via electrochemical reduction of a variety of aryl diazonium salts is described. The estimated degree of functionalization is as high as one out of every 20 carbons in the nanotubes bearing a functionalized moiety. The functionalizing moieties can be removed by heating in an argon atmosphere. Nanotubes derivatized with a 4 tert-butylbenzene moiety were found to possess significantly improved solubility in organic solvents. Functionalization of the nanotubes with a molecular system that has exhibited switching and memory behavior is shown. This represents the marriage of wire-like nanotubes with molecular electronic devices. PMID- 11439041 TI - Mechanism and activity of ruthenium olefin metathesis catalysts. AB - This report details the effects of ligand variation on the mechanism and activity of ruthenium-based olefin metathesis catalysts. A series of ruthenium complexes of the general formula L(PR(3))(X)(2)Ru=CHR(1) have been prepared, and the influence of the substituents L, X, R, and R(1) on the rates of phosphine dissociation and initiation as well as overall activity for olefin metathesis reactions was examined. In all cases, initiation proceeds by dissociative substitution of a phosphine ligand (PR(3)) with an olefinic substrate. All of the ligands L, X, R, and R(1) have a significant impact on initiation rates and on catalyst activity. The origins of the observed substituent effects as well as the implications of these studies for the design and implementation of new olefin metathesis catalysts and substrates are discussed in detail. PMID- 11439042 TI - Intermediate in beta-lactam hydrolysis catalyzed by a dinuclear zinc(II) complex: relevance to the mechanism of metallo-beta-lactamase. AB - Inactivation of beta-lactam antibiotics by metallo-beta-lactamase enzymes is a well-recognized pathway of antibiotic resistance in bacteria. As part of extensive mechanistic studies, the hydrolysis of a beta-lactam substrate nitrocefin (1) catalyzed by dinuclear zinc(II) model complexes was investigated in nonaqueous solutions. The initial step involves monodentate coordination of the nitrocefin carboxylate group to the dizinc center. The coordinated substrate is then attacked intramolecularly by the bridging hydroxide to give a novel intermediate (2') characterized by its prominent absorbance maximum at 640 nm, which affords a blue color. The NMR and IR spectroscopic data of 2' are consistent with it being zinc(II)-bound N-deprotonated hydrolyzed nitrocefin that forms from the tetrahedral intermediate upon C-N bond cleavage. Protonation of the leaving group is the rate-limiting step in DMSO solution and occurs after the C-N bond-breaking step. Addition of strong acids results in rapid conversion of 2' into hydrolyzed nitrocefin (3). The latter can be converted back to the blue species (2') upon addition of base. The low pK(a) value for the amino group in hydrolyzed nitrocefin is explained by its involvement in extended conjugation and by coordination to zinc(II). The blue intermediate (2') in the model system resembles well that in the enzymatic system, judging by its optical properties. The greater stability of the intermediate in the model, however, allowed its characterization by (13)C NMR and infrared, as well as electronic, spectroscopy. PMID- 11439043 TI - Molecular structures and magnetic resonance spectroscopic investigations of highly distorted six-coordinate low-spin iron(III) porphyrinate complexes. AB - Three bis-axially ligated complexes of iron(III) octaethyltetraphenylporphyrin, (OETPP)Fe(III), have been prepared, which are low-spin complexes, each with two axial nitrogen-donor ligands (N-methylimidazole (N-MeIm), 4 (dimethylamino)pyridine (4-NMe(2)Py), and 2-methylimidazole (2-MeImH)). The crystal and molecular structure of the bis-(2-MeImH) complex shows the macrocycle to be in a saddled conformation, with the ligands in perpendicular planes aligned at 14 degrees to the porphyrin nitrogens so as to relieve the steric interaction between the 2-methyl groups and the porphyrin. The Fe-N(por) bond lengths are typical of nonplanar six-coordinate low-spin Fe(III) complexes, while the axial Fe-N(ax) bond lengths are substantially longer than those of [(TPP)Fe(2 MeImH)(2)](+) (2.09(2) A as compared to 2.015(4) and 2.010(4) A). The crystal and molecular structure of the bis-(4-NMe(2)Py) complex also shows the macrocycle to be in a mainly saddled conformation, but with a significant ruffled component. As a result, the average Fe-N(por) bonds are significantly shorter (1.951 A as compared to 1.974 A) than those of the bis-(2-MeImH) complex. One ligand is aligned at 9 degrees to two trans porphyrin nitrogens, while the other is at 79 degrees to the same porphyrin nitrogens, producing a dihedral angle of 70 degrees between the ligand planes. The EPR spectrum of this complex, like that of the bis (2-MeImH) complex, is of the "large g(max)" type, with g(max) = 3.29 and 3.26, respectively. However, in frozen CD(2)Cl(2), [(OETPP)Fe(N-MeIm)(2)](+) exhibits both "large g(max)" and normal rhombic signals, suggesting the presence of both "perpendicular" and "parallel" ligand orientations. The 1- and 2D (1)H NMR spectra of each of these complexes, as well as the chloroiron(III) starting material, were investigated as a function of temperature. The COSY and NOESY/EXSY spectra of the chloride complex are consistent with the expected J-coupling and saddle inversion dynamics, respectively. Complete spectral assignments for the bis-(N-MeIm) and -(4-NMe(2)Py) complexes have been made using 2D (1)H NMR techniques. In each case, the number of resonances due to methylene (two) and phenyl protons (one each) is consistent with D(2)(d)() symmetry, and therefore an effective perpendicular orientation of the axial ligands on the time scale of the NMR experiments. The temperature dependences of the (1)H resonances of these complexes show significant deviations from Curie behavior, and also evidence of extensive ligand exchange and rotation. Spectral assignment of the eight methylene resonances of the bis-(2-MeImH) complex to the four ethyl groups was possible through the use of 2D (1)H NMR techniques. The complex is fluxional, even at -90 degrees C, and ROESY data suggest that the predominant process is saddle inversion accompanied by simultaneous rotation of the axial ligands. Saddle inversion becomes slow on the 2D NMR time scale as the temperature is lowered in the ligand order of N-MeIm > 4-NMe(2)Py > 2-MeImH, probably due mainly to progressive destabilization of the ground state rather than progressive stabilization of the transition state of the increasingly "hindered" bis-ligand complexes. PMID- 11439044 TI - Unexpected selectivities in C-H activations of toluene and p-xylene at cationic platinum(II) diimine complexes. New mechanistic insight into product-determining factors. AB - The C-H activation of toluene and p-xylene at cationic Pt(II) diimine complexes (N-N)Pt(CH(3))(H(2)O)(+)BF(4)(-) (N-N = Ar-N=CMe-CMe=N-Ar; 1(BF(4)(-)), N(f) N(f), Ar = 3,5-(CF(3))(2)C(6)H(3)); 2(BF(4)(-)), N'-N', Ar = 2,6 (CH(3))(2)C(6)H(3)) has been investigated. The reactions were performed at ambient temperature in 2,2,2-trifluoroethanol (TFE), and after complete conversion of the starting material to mixtures of Pt-aryl/Pt-benzyl complexes and methane, acetonitrile was added to trap the products as more stable acetonitrile adducts. In the reactions with toluene, the relative amounts of products resulting from aromatic C-H activation were found to decrease in the order (N-N)Pt(m-tolyl)(NCMe)(+) > (N-N)Pt(p-tolyl)(NCMe)(+) > (N-N)Pt(o tolyl)(NCMe)(+) for both 1 and 2. Unlike the reaction at 1, significant amounts of the benzylic activation product (N'-N')Pt(benzyl)(NCMe)(+) were concurrently formed in the C-H activation of toluene at 2. The C-H activation of p-xylene revealed an even more remarkable difference between 1 and 2. Here, the product ratios of (N-N)Pt(xylyl)(NCMe)(+) and (N-N)Pt(p-methylbenzyl)(NCMe)(+) were found to be 90:10 and 7:93 for reactions at 1 and 2, respectively. The elimination of toluene from (N(f)-N(f))Pt(Tol)(2) species (3a-c; a, Tol = o-tolyl; b, Tol = m tolyl; c, Tol = p-tolyl) after protonolysis with 1 equiv of HBF(4) was investigated. Most notably, protonation in neat TFE followed by addition of acetonitrile gave a 77:23 mixture of (N(f)-N(f))Pt(m-tolyl)(NCMe)(+) (4b) and (N(f)-N(f))Pt(p-tolyl)(NCMe)(+) (4c) from all three isomeric bis(tolyl) complexes 3a-c. The presence of acetonitrile during the protonation reactions resulted in considerably less isomerization. This behavior is explained by an associative mechanism for the product-determining displacement of toluene by the solvent. For the C-H activation reactions, our findings suggest the existence of a dynamic equilibrium between the isomeric intermediates (N-N)Pt(aryl)(CH(4))(+) (aryl = tolyl/benzyl from 1; xylyl/p-methylbenzyl from 2). The observed selectivities might then be explained by steric and electronic effects in the pentacoordinate transition-state structures for the solvent-induced associative elimination of methane from these intermediates. PMID- 11439045 TI - Decay of the peroxide intermediate in laccase: reductive cleavage of the O-O bond. AB - Laccase is a multicopper oxidase that contains four Cu ions, one type 1, one type 2, and a coupled binuclear type 3 Cu pair. The type 2 and type 3 centers form a trinuclear Cu cluster that is the active site for O(2) reduction to H(2)O. To examine the reaction between the type 2/type 3 trinuclear cluster and dioxygen, the type 1 Cu was removed and replaced with Hg(2+), producing the T1Hg derivative. When reduced T1Hg laccase is reacted with dioxygen, a peroxide intermediate (P) is formed. The present study examines the kinetics and mechanism of formation and decay of P in T1HgLc. The formation of P was found to be independent of pH and did not involve a kinetic solvent isotope effect, indicating that no proton is involved in the rate-determining step of formation of P. Alternatively, pH and isotope studies on the decay of P revealed that a proton enhances the rate of decay by 10-fold at low pH. This process shows an inverse k(H)/k(D) kinetic solvent isotope effect and involves protonation of a nearby residue that assists in catalysis, rather than direct protonation of the peroxide. Decay of P also involves a significant oxygen isotope effect (k(16)O(2)/k(18)O(2)) of 1.11 +/- 0.05, indicating that reductive cleavage of the O-O bond is the rate-determining step in the decay of P. The activation energy for this process was found to be approximately 9.0 kcal/mol. The exceptionally slow rate of decay of P is explained by the fact that this process involves a 1e( ) reductive cleavage of the O-O bond and there is a large Franck-Condon barrier associated with this process. Alternatively, the 2e(-) reductive cleavage of the O-O bond has a much larger driving force which minimizes this barrier and accelerates the rate of this reaction by approximately 10(7) in the native enzyme. This large difference in rate for the 2e(-) versus 1e(-) process supports a molecular mechanism for multicopper oxidases in which O(2) is reduced to H(2)O in two 2e(-) steps. PMID- 11439046 TI - Electron-rich rods as building blocks for Sb strips and Te sheets. AB - We analyze the bonding in a number of networks of heavy main group elements comprised of finite-length linear chains fused at right angles. Isolated linear chain building blocks may be understood easily by analogy with three-orbital four electron "hypervalent" bonding picture in such molecules as I(3)(-) and XeF(2). After deriving the appropriate electron-counting rules for such linear units, we proceed in an aufbau to fuse these chains into simple (and not so simple) infinite networks. It is proposed that (a) infinite Sb(3) ribbons of vertex sharing squares are stable for an electron count of 20 electrons per three atoms (i.e., ); (b) sidewise fused Sb double ribbons are stable for an electron count of 38 electrons per six atoms (i.e., ); (c) Sb(4) strips cut from a square lattice are stable at the electron count of 24 electrons per four atoms (i.e., ); (d) Te(6) defect square sheets are stable at the electron count of 40 electrons per six atoms (i.e., ). The electronic structures of the solid-state compounds containing these networks, namely La(12)Mn(2)Sb(30), alpha-ZrSb(2), beta-ZrSb(2), Cs(3)Te(22), and Cs(4)Te(28), are elaborated. We propose preferred electron counts for two hypothetical Sb ribbons derived from the Sb(3) ribbon in La(12)Mn(2)Sb(30). A possibility of geometry distortion modulation by excess charge in lattices comprised of even-membered linear units is suggested. PMID- 11439047 TI - Interactions of hydrogen with CeO(2). AB - The interactions of hydrogen with ceria have been the subject of several experimental studies, but whether hydrogen atoms enter the bulk or are merely chemisorbed on the surfaces is disputed. We combine first-principles calculations with thermodynamic arguments and conclude that the uptake of small amounts of hydrogen by ceria is spontaneous below 665 K. The calculations also predict that hydrogen atoms within the bulk form hydroxyl groups and slightly expand the lattice. The predicted hydrogen content, OH vibrational frequency, and ceria lattice expansion upon hydrogen uptake are consistent with available data. PMID- 11439048 TI - Nonisomerizable non-retinal chromophores initiate light-induced conformational alterations in bacterioopsin. AB - The photoactivation of retinal proteins is usually interpreted in terms of C=C photoisomerization of the retinal moiety, which triggers appropriate conformational changes in the protein. In this work several dye molecules, characterized by a completely rigid structure in which no double-bond isomerization is possible, were incorporated into the binding site of bacteriorhodopsin (bR). Using a light-induced chemical reaction of a labeled EPR probe, it was observed that specific conformational alterations in the protein are induced following light absorption by the dye molecules occupying the binding site. The exact nature of these changes and their relationship to those occurring in the bR photocycle are still unclear. Nevertheless, their occurrence proves that C=C or C=NH(+) isomerization is not a prerequisite for protein conformational changes in a retinal protein. More generally, we show that conformational changes, leading to changes in reactivity, may be induced in proteins by optical excitation of simple nonisomerizable dyes located in the macromolecular matrix. PMID- 11439049 TI - Charge separation in a novel artificial photosynthetic reaction center lives 380 ms. AB - An extremely long-lived charge-separated state has been achieved successfully using a ferrocene-zincporphyrin-freebaseporphyrin-fullerene tetrad which reveals a cascade of photoinduced energy transfer and multistep electron transfer within a molecule in frozen media as well as in solutions. The lifetime of the resulting charge-separated state (i.e., ferricenium ion-C(60) radical anion pair) in a frozen benzonitrile is determined as 0.38 s, which is more than one order of magnitude longer than any other intramolecular charge recombination processes of synthetic systems, and is comparable to that observed for the bacterial photosynthetic reaction center. Such an extremely long lifetime of the tetrad system has been well correlated with the charge-separated lifetimes of two homologous series of porphyrin-fullerene dyad and triad systems. PMID- 11439050 TI - The mechanism of 1,2-addition of disilene and silene. 1. Water and alcohol addition. AB - The mechanism of 1,2-addition reactions of water, methanol, and trifluoromethanol to Si=Si, Si=C, and C=C bonds has been investigated by ab initio quantum chemical methods. Geometries and relative energies of the stationary points and all the transition states were determined using the MP2/6-311++G(d,p), B3LYP/6 311++G(d,p), and CBS-Q levels of theory. The investigated reactions can be characterized by two main thermodynamical profiles. The type in which the reagent molecule attacks a carbon atom is moderately exothermic, with a high activation barrier. The second type, in which water or alcohol attacks a silicon, is strongly exothermic, with a small activation energy. At the early stage of all the reactions, a weakly bonded initial complex is found which determines the further mechanism of the reaction. On the basis of the HOMO, LUMO, and Laplacian of electron distribution of disilene and silene, several mechanisms have been assumed, depending on the substrate (disilene, substituted disilene, silene, or ethene) and the reagent (water, methanol, or trifluoromethanol). The reaction diagrams and proposed mechanisms explain the experimentally found regioselectivity and diastereoselectivity well. PMID- 11439051 TI - Stability of chiral domains produced by adsorption of tartaric acid isomers on the Cu(110) surface: a periodic density functional theory study. AB - In the present work the interaction of different bitartrate isomers on the Cu(110) surface has been investigated systematically by using the Vienna Ab initio Simulation Package (VASP), which performs periodical density functional theory (DFT) calculations. Among all bitartrate isomers the R,R-configuration is the most stable under the (3 1, 1 2) domain on the Cu surface. Its optical isomer, the S,S-bitartrate, is 10 kJ mol(-)(1) less stable in the same domain. This energy difference is sufficient to produce the distinct chiral assemblies observed after the adsorption of each optical isomer on the Cu surface. The calculations also showed that these domains are not formed due to intermolecular H-bonds, in contrast with the previous proposal by Raval et al.(Nature 2000, 23, 376). In fact, there is a formation of optimal intramolecular H-bonds in the chemisorption structures. A favorable packing orientation is also needed for the respective chiral domains. For instance, the S,S-configuration suffers from a destabilizing packing energy of 21 kJ mol(-)(1) under the same domain, due to a short contact between the H atoms of the hydroxy groups. These intramolecular H bonds cause also some distortions on the bitartrate molecule, which appear to be dependent on the relative position of the alpha-hydroxy groups. The stability of the extended asymmetric domains, when the surface is modified by a chiral additive, might have important consequences for understanding and optimizing the properties of enantioselective heterogeneous catalysts. PMID- 11439052 TI - Kinetics of histidine deligation from the heme in GuHCl-unfolded Fe(III) cytochrome C studied by a laser-induced pH-jump technique. AB - We have developed an instrumental setup that uses transient absorption to monitor protein folding/unfolding processes following a laser-induced, ultrafast release of protons from o-nitrobenzaldehyde. The resulting increase in [H(+)], which can be more than 100 microM, is complete within a few nanoseconds. The increase in [H(+)] lowers the pH of the solution from neutrality to approximately 4 at the highest laser pulse energy used. Protein structural rearrangements can be followed by transient absorption, with kinetic monitoring over a broad time range (approximately 10 ns to 500 ms). Using this pH-jump/transient absorption technique, we have examined the dissociation kinetics of non-native axial heme ligands (either histidine His26 or His33) in GuHCl-unfolded Fe(III) cytochrome c (cyt c). Deligation of the non-native ligands following the acidic pH-jump occurs as a biexponential process with different pre-exponential factors. The pre exponential factors markedly depend on the extent of the pH-jump, as expected from differences in the pK(a) values of His26 and His33. The two lifetimes were found to depend on temperature but were not functions of either the magnitude of the pH-jump or the pre-pulse pH of the solution. The activation energies of the deligation processes support the suggestion that GuHCl-unfolded cyt c structures with non-native histidine axial ligands represent kinetic traps in unfolding. PMID- 11439053 TI - Formation of a phosphorus-phosphorus bond by successive one-electron reductions of a two-phosphinines-containing macrocycle: crystal structures, EPR, and DFT investigations. AB - Chemical and electrochemical reductions of the macrocycle 1 lead to the formation of a radical monoanion anion [1](*)(-) whose structure has been studied by EPR in liquid and frozen solutions. In accord with experimental (31)P hyperfine tensors, DFT calculations indicate that, in this species, the unpaired electron is mainly localized in a bonding sigma P-P orbital. Clearly, a one-electron bond (2.763 A) was formed between two phosphorus atoms which, in the neutral molecule, were 3.256 A apart (crystal structure). A subsequent reduction of this radical anion gives rise to the dianion [1](2)(-) which could be crystallized by using, in the presence of cryptand, Na naphthalenide as a reductant agent. As shown by the crystal structure, in [1](2)(-), the two phosphinine moieties adopt a phosphacyclohexadienyl structure and are linked by a P-P bond whose length (2.305(2) A) is only slightly longer than a usual P-P bond. When the phosphinine moieties are not incorporated in a macrocycle, no formation of any one-electron P P bond is observed: thus, one-electron reduction of 3 with Na naphthalenide leads to the EPR spectrum of the ion pair [3](*)(-) Na(+); however, at high concentration, these ion pairs dimerize, and, as shown by the crystal structure of [(3)(2)](2)(-)[(Na(THF)(2))(2)](2+) a P-P bond is formed (2.286(2) A) between two phosphinine rings which adopt a boat-type conformation, the whole edifice being stabilized by two carbon-sodium-phosphorus bridges. PMID- 11439054 TI - Theoretical studies on alkyne addition to molybdenum alkylidenes: the preference of alpha- and beta-additions. AB - The addition of alkynes HC=CR to Mo(NH)(CH(2))(OR')(2) (R = H, Me, Ph; R' = CH(3), CF(3)) has been studied with both ab initio molecular orbital and density functional calculations. Geometry optimizations were carried out with the HF/3 21G, HF/HW3, and B3LYP/HW3 methods. The transition structures for these addition reactions are in distorted trigonal bipyramidal geometries, similar to those of alkene additions. The calculated activation enthalpy for HC=CH addition to Mo(NH)(CH(2))(OR')(2) is about 10.3 kcal/mol for R' = CH(3) and about 2.3 kcal/mol for R' = CF(3), indicating a significant preference for acetylene addition to Mo(NH)(CH(2))(OCF(3))(2) over Mo(NH)(CH(2))(OCH(3))(2). These barriers are higher than those of the corresponding ethylene addition by about 2 4 kcal/mol, even though the reaction of acetylene is much more exothermic. The alpha-addition of HC=CR (R = Me, Ph) is found to be considerably more favorable than the beta-addition to Mo(NH)(CH(2))(OR')(2). Interestingly, the alpha addition has a lower activation energy, while the beta-addition has a higher activation energy, compared to that of the parent acetylene addition. Thus, alpha addition is intrinsically favored over beta-addition by over 4 kcal/mol. This preference is reduced by solvent effect. All these can be explained by a destabilizing interaction between the nonreacting pi-orbital of alkyne and one of the lone pairs on the imido nitrogen. The steric effect of the bulky ligands in the real catalysts is also investigated qualitatively by the PM3 method. These studies give results in good accord with the experimentally observed regioselectivity. PMID- 11439055 TI - Single two-electron transfers vs successive one-electron transfers in polyconjugated systems illustrated by the electrochemical oxidation and reduction of carotenoids. AB - Examination of cyclic voltammetric responses reveals that inversion of the standard potentials of the first and second electron transfers occurs in the oxidation of beta-carotene and 15,15'-didehydro-beta-carotene (but not in their reduction) as well as in the reduction of canthaxanthin (but not in its oxidation). The factors that control potential inversion in these systems, and more generally in symmetrical molecules containing conjugated long chains, are investigated by quantum chemical calculations. Two main interconnected effects emerge. One is the localization of the charges in the di-ion toward the ends of the molecule at a large distance from one another, thus minimizing Coulombic repulsion. The same effect favors the solvation of the di-ion providing additional stabilization. In contrast, the charge in the ion radical is delocalized over the whole molecular framework, thus disfavoring its stabilization by interaction with the solvent. The combination of the two solvation effects allows potential inversion to occur as opposed to the case where the two electrophores are linked by a saturated bridge where potential inversion cannot occur. Localization of the charges in the di-ion, and thus potential inversion, is favored by the presence of electron-accepting terminal groups for reductions (as the two carbonyl groups in canthaxanthin) and of hole accepting terminal groups for oxidations (as in beta-carotene). PMID- 11439056 TI - Solution structure, dimerization, and dynamics of a lipophilic alpha/3(10) helical, C alpha-methylated peptide. Implications for folding of membrane proteins. AB - The solution structure and the dimerization behavior of the lipophilic, highly C(alpha)-methylated model peptide, mBrBz-Iva(1)-Val(2)-Iva(3)-(alphaMe)Val(4) (alphaMe)Phe(5)-(alphaMe)Val(6)-Iva(7)-NHMe, was studied by NMR spectroscopy and molecular dynamics simulations. The conformational analysis resulted in a right handed 3(10)/alpha-helical equilibrium fast on the NMR time scale with a slight preference for the alpha-helical conformation. The NOESY spectrum showed intermolecular NOEs due to an aggregation of the heptapeptide. In addition, temperature-dependent diffusion measurements were performed to calculate the hydrodynamic radius. All these findings are consistent with an antiparallel side by-side dimerization. The structure of the dimeric peptide was calculated with a simulated annealing strategy. The lipophilic dimer is held together by favorable van der Waals interactions in the sense of a bulge fitting into a groove. The flexibility of the helical conformations concerning an alpha/3(10)-helical equilibrium is shown in a 3 ns molecular dynamics simulation of the resulting dimeric structure. Both overall helical structures of each monomer and the antiparallel mode of dimerization are stable. However, transitions were seen of several residues from a 3(10)-helical into an alpha-helical conformation and vice versa. Hence, this peptide represents a good model in which two often-discussed aspects of hierarchical transmembrane protein folding are present: i <-- i + 3 and i <-- i + 4 local H-bonding interactions cause a specific molecular shape which is then recognized as attractive by other surrounding structures. PMID- 11439057 TI - Corannulene as a Lewis base: computational modeling of protonation and lithium cation binding. AB - A computational modeling of the protonation of corannulene at B3LYP/6 311G(d,p)//B3LYP/6-311G(d,p) and of the binding of lithium cations to corannulene at B3LYP/6-311G(d,p)//B3LYP/6-31G(d,p) has been performed. A proton attaches preferentially to one carbon atom, forming a sigma-complex. The isomer protonated at the innermost (hub) carbon has the best total energy. Protonation at the outermost (rim) carbon and at the intermediate (bridgehead rim) carbon is less favorable by ca. 2 and 14 kcal mol(-)(1), respectively. Hydrogen-bridged isomers are transition states between the sigma-complexes; the corresponding activation energies vary from 10 to 26 kcal mol(-)(1). With an empirical correction obtained from calculations on benzene, naphthalene, and azulene, the best estimate for the proton affinity of corannulene is 203 kcal mol(-)(1). The lithium cation positions itself preferentially over a ring. There is a small energetic preference for the 6-ring over the 5-ring binding (up to 2 kcal mol(-)(1)) and of the convex face over the concave face (3-5 kcal mol(-)(1)). The Li-bridged complexes are transition states between the pi-face complexes. Movement of the Li(+) cation over either face is facile, and the activation energy does not exceed 6 kcal mol(-)(1) on the convex face and 2.2 kcal mol(-)(1) on the concave face. In contrast, the transition of Li(+) around the corannulene edge involves a high activation barrier (24 kcal mol(-)(1) with respect to the lowest energy pi face complex). An easier concave/convex transformation and vice versa is the bowl to-bowl inversion with an activation energy of 7-12 kcal mol(-)(1). The computed binding energy of Li(+) to corannulene is 44 kcal mol(-)(1). Calculations of the (7)Li NMR chemical shifts and nuclear independent chemical shifts (NICS) have been performed to analyze the aromaticity of the corannulene rings and its changes upon protonation. PMID- 11439058 TI - Long-distance radical cation migration in Z-form DNA. PMID- 11439059 TI - First divergent strategy using two AB(2) unprotected monomers for the rapid synthesis of dendrimers. PMID- 11439060 TI - Activation of a terminal carboxylic acid by an internal oxazole: a novel synthesis of macrocyclodepsipeptide. PMID- 11439061 TI - Convergent synthesis of polycyclic ethers via the intramolecular allylation of alpha-acetoxy ethers and subsequent ring-closing metathesis: synthesis of the CDEFG ring system of gambierol. PMID- 11439062 TI - Norbornyne: a cycloalkyne reacting like a dicarbene. PMID- 11439063 TI - DFT study of the cycloaddition reactions of strained alkynes. PMID- 11439064 TI - Oxygenation of phenols to catechols by a (mu-eta 2:eta 2-peroxo)dicopper(II) complex: mechanistic insight into the phenolase activity of tyrosinase. PMID- 11439065 TI - Enhancement of enantiomeric excess by ligand distortion. PMID- 11439066 TI - Mapping the mitomycin biosynthetic pathway by functional analysis of the MitM aziridine N-methyltransferase. PMID- 11439067 TI - Photoinduced electron transfer and photovoltaic devices of a conjugated polymer with pendant fullerenes. PMID- 11439069 TI - Stereochemistry of thermal vinylcyclobutane-to- cyclohexene rearrangements of cis (1S,2R)- and trans-(1S,2S)-1-(E)-propenyl-2-methylcyclobutanes. PMID- 11439068 TI - A diastereoselective metal-catalyzed [2+2] cycloaddition of bis-enomes. PMID- 11439070 TI - Electron spin catalysis by xenon in an enzyme. PMID- 11439071 TI - Modeling Rieske dioxygenases: the first example of iron-catalyzed asymmetric cis dihydroxylation of olefins. PMID- 11439073 TI - Tuning of superquenching in layered and mixed fluorescent polyelectrolytes. PMID- 11439072 TI - Total synthesis of the Kopsia lapidilecta alkaloid (+/-)-lapidilectine B. PMID- 11439074 TI - Exposing asymmetry between monomers in Alzheimer's amyloid fibrils via reductive alkylation of lysine residues. PMID- 11439075 TI - Single-chain versus dimeric protein folding: thermodynamic and kinetic consequences of covalent linkage. PMID- 11439076 TI - Chemical vapor deposition of diamond: an in situ study by vibrational spectroscopy. PMID- 11439077 TI - Oligonucleotide detection using angle-dependent light scattering and fractal dimension analysis of gold-DNA aggregates. PMID- 11439078 TI - Softened hydrophobic attraction between macroscopic surfaces in relative motion. PMID- 11439079 TI - Homochiral G-quadruplexes with Ba2+ but not with K+: the cation programs enantiomeric self-recognition. PMID- 11439080 TI - Use of biomimetic diversity-oriented synthesis to discover galanthamine-like molecules with biological properties beyond those of the natural product. PMID- 11439081 TI - Molecular recognition of a RNA:DNA hybrid structure. PMID- 11439082 TI - Neuroendocrine secretory protein 7B2: structure, expression and functions. AB - 7B2 is an acidic protein residing in the secretory granules of neuroendocrine cells. Its sequence has been elucidated in many phyla and species. It shows high similarity among mammals. A Pro-Pro-Asn-Pro-Cys-Pro polyproline motif is its most conserved feature, being carried by both vertebrate and invertebrate sequences. It is biosynthesized as a precursor protein that is cleaved into an N-terminal fragment and a C-terminal peptide. In neuroendocrine cells, 7B2 functions as a specific chaperone for the proprotein convertase (PC) 2. Through the sequence around its Pro-Pro-Asn-Pro-Cys-Pro motif, it binds to an inactive proPC2 and facilitates its transport from the endoplasmic reticulum to later compartments of the secretory pathway where the zymogen is proteolytically matured and activated. Its C-terminal peptide can inhibit PC2 in vitro and may contribute to keep the enzyme transiently inactive in vivo. The PC2-7B2 model defines a new neuroendocrine paradigm whereby proteolytic activation of prohormones and proneuropeptides in the secretory pathway is spatially and temporally regulated by the dynamics of interactions between converting enzymes and their binding proteins. Interestingly, unlike PC2-null mice, which are viable, 7B2-null mutants die early in life from Cushing's disease due to corticotropin ('ACTH') hypersecretion by the neurointermediate lobe, suggesting a possible involvement of 7B2 in secretory granule formation and in secretion regulation. The mechanism of this regulation is yet to be elucidated. 7B2 has been shown to be a good marker of several neuroendocrine cell dysfunctions in humans. The possibility that anomalies in its structure and expression could be aetiological causes of some of these dysfunctions warrants investigation. PMID- 11439083 TI - Variation in aspects of cysteine proteinase catalytic mechanism deduced by spectroscopic observation of dithioester intermediates, kinetic analysis and molecular dynamics simulations. AB - The possibility of a slow post-acylation conformational change during catalysis by cysteine proteinases was investigated by using a new chromogenic substrate, N acetyl-Phe-Gly methyl thionoester, four natural variants (papain, caricain, actinidin and ficin), and stopped-flow spectral analysis to monitor the pre steady state formation of the dithioacylenzyme intermediates and their steady state hydrolysis. The predicted reversibility of acylation was demonstrated kinetically for actinidin and ficin, but not for papain or caricain. This difference between actinidin and papain was investigated by modelling using QUANTA and CHARMM. The weaker binding of hydrophobic substrates, including the new thionoester, by actinidin than by papain may not be due to the well-known difference in their S2-subsites, whereby that of actinidin in the free enzyme is shorter due to the presence of Met211. Molecular dynamics simulation suggests that during substrate binding the sidechain of Met211 moves to allow full access of a Phe sidechain to the S2-subsite. The highly anionic surface of actinidin may contribute to the specificity difference between papain and actinidin. During subsequent molecular dynamics simulations the P1 product, methanol, diffuses rapidly (over<8 ps) out of papain and caricain but 'lingers' around the active centre of actinidin. Uniquely in actinidin, an Asp142-Lys145 salt bridge allows formation of a cavity which appears to constrain diffusion of the methanol away from the catalytic site. The cavity then undergoes large scale movements (over 4.8 A) in a highly correlated manner, thus controlling the motions of the methanol molecule. The changes in this cavity that release the methanol might be those deduced kinetically. PMID- 11439084 TI - Metalloprotease-disintegrin ADAM 12 interacts with alpha-actinin-1. AB - ADAM 12, a member of the ADAM family of proteins (containing A Disintegrin And Metalloprotease domain), has been implicated in differentiation and fusion of myoblasts. While the extracellular domain of ADAM 12 contains an active metalloprotease and a region involved in cell adhesion, the function of the cytoplasmic tail of ADAM 12 has been less clear. Here we show that the cytoplasmic domain of ADAM 12 interacts in vitro and in vivo with alpha-actinin 1, an actin-binding and cross-linking protein. Green fluorescent protein fused to ADAM 12 cytoplasmic domain co-localizes with alpha-actinin-1-containing actin stress fibres in C2C12 cells. The interaction between ADAM 12 and alpha-actinin-1 is direct and involves the 58-amino acid C-terminal fragment of ADAM 12 and the 27 kDa N-terminal domain of alpha-actinin-1. Consistently, expression of the 27 kDa fragment of alpha-actinin-1 in C2C12 cells using a mitochondrial targeting system results in recruitment of the co-expressed ADAM 12 cytoplasmic domain to the mitochondrial surface. Moreover, alpha-actinin-1 co-purifies with a transmembrane, His6-tagged form of ADAM 12 expressed in C2C12 myoblasts, indicating that the transmembrane ADAM 12 forms a complex with alpha-actinin-1 in vivo. These results indicate that the actin cytoskeleton may play a critical role in ADAM 12-mediated cell-cell adhesion or cell signalling during myoblast differentiation and fusion. PMID- 11439085 TI - Functional coupling between the caffeine/ryanodine-sensitive Ca2+ store and mitochondria in rat aortic smooth muscle cells. AB - We investigated the role of mitochondria in the agonist-induced and/or caffeine induced Ca2+ transients in rat aortic smooth muscle cells. We explored the possibility that proliferation modulates the coupling between mitochondria and endoplasmic reticulum. Ca2+ transients induced by either ATP or caffeine were measured in presence or absence of drugs interfering with mitochondrial activity in freshly dissociated cells (day 1) and in subconfluent primary culture (day 12). We found that the mitochondrial inhibitors, rotenone or carbonyl cyanide m chlorophenylhydrazone, as well as the permeability transition pore inhibitor, cyclosporin A, had no effect on the ATP-induced Ca2+ transient at either day 1 or day 12, but prevented caffeine-induced cytosolic Ca2+ increase at day 12 but not at day 1. Close connections between ryanodine receptors and mitochondria were observed at both day 1 and 12. Thapsigargin (TG) prevented ATP- and caffeine induced Ca2+ transients at day 1. At day 12, where only 50% of the cells were sensitive to caffeine, TG did not prevent the caffeine-induced Ca2+ transient, and prevented ATP-induced Ca2+ transient in only half of the cells. Together, these data demonstrate that rat aortic smooth muscle cells at day 1 have an ATP- and caffeine-sensitive pool, which is functionally independent but physically closely linked to mitochondria and totally inhibited by TG. At day 12, we propose the existence of two cell populations: half contains IP3 receptors and TG sensitive Ca2+ pumps only; the other half contains, in addition to the IP3 sensitive pool independent from mitochondria, a caffeine-sensitive pool. This latter pool is linked to mitochondria through the permeability transition pore and is refilled by both TG-sensitive and insensitive mechanisms. PMID- 11439086 TI - Macrophage migration inhibitory factor of the parasitic nematode Trichinella spiralis. AB - cDNAs were obtained for macrophage migration-inhibitory factor (MIF)/L-dopachrome methyl ester tautomerase homologues from the parasitic nematodes Trichinella spiralis (TsMIF) and Trichuris trichiura (TtMIF). The translated sequences, which were partly confirmed by sequencing of proteolytic fragments, show 42 and 44% identity respectively with human or mouse MIF, and are shorter by one C-terminal residue. Unlike vertebrate MIF and MIF homologues of filarial nematodes, neither TsMIF nor TtMIF contain cysteine residues. Soluble recombinant TsMIF, expressed in Escherichia coli showed secondary structure (by CD spectroscopy) and quaternary structure (by light-scattering and gel filtration) similar to that of the trimeric mammalian MIFs and D-dopachrome tautomerase. The catalytic specificity of recombinant TsMIF in the ketonization of phenylpyruvate (1.4x10(6) M(-1) x s(-1)) was comparable with that of human MIF, while that of p hydroxyphenylpyruvate (9.1x10(4) M(-1) x s(-1)) was 71-fold lower. TsMIF showed high specificity in tautomerization of the methyl ester of L-dopachrome compared with non-esterified L-dopachrome (>87000-fold) and a high kcat (approximately 4x10(4) s(-1). The crystal structure, determined to 1.65 A (1 A=0.1 nm), was generally similar to that of human MIF, but differed in the boundaries of the putative active-site pocket, which can explain the low activity towards p hydroxyphenylpyruvate. The central pore was blocked, but was continuous, with the three putative tautomerase sites. Recombinant TsMIF (5 ng/ml-5 pg/ml) inhibited migration of human peripheral-blood mononuclear cells in a manner similar to that shown by human MIF, but had no effect from 5 to 500 ng/ml on anti-CD3-stimulated murine T-cell proliferation. TsMIF was detected in supernatants of T. spiralis larvae cultured in vitro at 6 ng/ml (55 ng/mg total secreted protein). In conclusion TsMIF has structural, catalytic and cell-migration-inhibitory properties which indicate that it is partially orthologous to mammalian MIF. PMID- 11439087 TI - Site-directed removal of N-glycosylation sites in BST-1/CD157: effects on molecular and functional heterogeneity. AB - Cyclic ADP ribose (cADPR) is a novel second messenger that releases calcium from intracellular calcium stores, but works independently of inositol 1,4,5 trisphosphate. In mammals ADP-ribosyl cyclase function is found in two membrane proteins, CD38 and bone marrow stromal cell antigen 1 (BST-1)/CD157. These enzymes are exposed extracellularly and also possess cADPR hydrolase activity, but an intracellular soluble ADP-ribosyl cyclase has been reported in human T cells. Previously, a soluble form of BST-1/CD157 (sBST-1), which lacked the glycosylphosphatidylinositol-anchored portion, was expressed by a baculovirus insect-cell system. In this study, we have purified the sBST-1, and it migrated as two major bands by SDS/PAGE, suggesting that it is post-translationally modified. BST-1 contains four putative N-glycosylation sites. Tunicamycin treatment reduced sBST-1 expression in the culture medium, indicating that N glycosylation is essential for secretion. Site-directed mutagenesis was performed to generate sBST-1 mutants (N1-N4), each preserving a single N-glycosylation site. N1, N3 and N4 were well secreted into the medium, and were each detected as a single band. Although N3 and N4 retained the ADP-ribosyl cyclase activity, the cADPR-hydrolase activity was retained only in N4. We conclude that N glycosylation of sBST-1 facilitates the folding of the nascent polypeptide chain into a conformation that is conductive for intracellular transport and enzymic activity. Furthermore a crystal has been obtained using the N4 mutant, but not the wild-type sBST-1. Thus the artificial engineering of N-glycosylation sites could be an effective method to generate homogeneous material for structural studies. PMID- 11439088 TI - Identification and characterization of molecular interactions between glucose regulated proteins (GRPs) mortalin/GRP75/peptide-binding protein 74 (PBP74) and GRP94. AB - A heat-shock protein (hsp) 70 family member mortalin/glucose-regulated protein (GRP) 75/peptide-binding protein 74 (PBP74) has been localized to various cellular compartments including mitochondria, endoplasmic reticulum and cytoplasmic vesicles. Here we describe its interactions with an endoplasmic reticulum protein GRP94, a member of the hsp90 family of GRPs. Interactions were identified, confirmed and characterized by far-Western screening, in vivo reporter and co-immunoprecipitation assays. Interacting domains of the two proteins were also characterized by mutational analysis. Such interactions of these two GRPs may be important for function of either or both and therefore provide important information for further studies. PMID- 11439089 TI - Characterization of active-site residues in diadenosine tetraphosphate hydrolase from Lupinus angustifolius. AB - Site-directed mutagenesis has been used to characterize the functions of key amino acid residues in the catalytic site of the 'nudix' hydrolase, (asymmetrical) diadenosine 5',5"'-P1,P4-tetraphosphate (Ap4A) hydrolase (EC 3.6.1.17) from Lupinus angustifolius, the three-dimensional solution structure of which has recently been solved. Residues within the nudix motif, Gly-(Xaa)5-Glu (Xaa)7-Arg-Glu-Uaa-Xaa-(Glu)2-Xaa-Gly (where Xaa represents unspecified amino acids and Uaa represents the bulky aliphatic amino acids Ile, Leu or Val) conserved in 'nudix enzymes', and residues important for catalysis from elsewhere in the molecule, were mutated and the expressed proteins characterized. The results reveal a high degree of functional conservation between lupin asymmetric Ap4A hydrolase and the 8-oxo-dGTP hydrolase from Escherichia coli. Charged residues in positions equivalent to those that ligate an enzyme-bound metal ion in the E. coli 8-oxo-dGTP hydrolase [Harris, Wu, Massiah and Mildvan (2000) Biochemistry 39, 1655-1674] were shown to contribute to catalysis to similar extents in the lupin enzyme. Mutations E55Q, E59Q and E125Q all reduced kcat markedly, whereas mutations R54Q, E58Q and E122Q had smaller effects. None of the mutations produced a substantial change in the Km)for Ap4A, but several extensively modified the pH-dependence and fluoride-sensitivities of the hydrolase. It was concluded that the precisely positioned glutamate residues Glu 55, Glu-59 and Glu-125 are conserved as functionally significant components of the hydrolytic mechanism in both of these members of the nudix family of hydrolases. PMID- 11439090 TI - Temporal relationships between ceramide production, caspase activation and mitochondrial dysfunction in cell lines with varying sensitivity to anti-Fas induced apoptosis. AB - To clarify the chronology of events leading to anti-Fas-induced apoptosis, and the mechanisms of resistance to this death effector, we compared the response kinetics of three tumour cell lines that display varying sensitivity to anti-Fas (based on levels of apoptosis), in terms of ceramide release, mitochondrial function and the caspase-activation pathway. In the highly sensitive Jurkat cell line, early caspase-8 activation, observed from 2 h after treatment, was chronologically associated with an acute depletion of glutathione and the cleavage of caspase-3 and poly-ADP ribosyl polymerase (PARP), followed by a progressive fall in the mitochondrial transmembrane potential (Delta(psi)m), between 4 and 48 h after treatment. Ceramide levels began to increase 2 h after the addition of anti-Fas (with no increase during the first hour), and increased continuously to 640% of control cells at 48 h. In the moderately sensitive SCC61 adherent cells, comparable results were observed, though with lower levels of ceramide and a delay in the response kinetics, with apoptotic cells becoming flotant. Finally, despite early cleavage of caspase-8 at 2 h, and a sustained level of activation until 48 h, no apoptotic response was observed in anti-Fas resistant SQ20B cells. This was confirmed by a lack of ceramide generation and mitochondrial changes, and by the absence of any detectable cleavage of caspase-3 or PARP. Inhibition of caspase processing, and amplification of endogenous ceramide signalling by pharmacological agents, allowed us to establish the order of cellular events, locating ceramide release after caspase-8 activation and before caspase-3 activation, and demonstrating a direct involvement for ceramide release in mitochondrial dysfunction. Furthermore, these experiments provide strong arguments for the role of endogenous ceramide as a key executor of apoptosis, rather than as a consequence of membrane alterations. PMID- 11439091 TI - Phenotype-genotype relationships in peroxisome biogenesis disorders of PEX1 defective complementation group 1 are defined by Pex1p-Pex6p interaction. AB - The peroxisome biogenesis disorders (PBDs), including Zellweger syndrome (ZS), neonatal adrenoleucodystrophy (NALD) and infantile Refsum disease (IRD), are fatal autosomal recessive diseases caused by impaired peroxisome biogenesis, of which 12 genotypes have been reported. ZS patients manifest the severest clinical and biochemical abnormalities, whereas those with NALD and IRD show less severity and the mildest features respectively. We have reported previously that temperature-sensitive peroxisome assembly is responsible for the mildness of the clinical features of IRD. PEX1 is the causative gene for PBDs of complementation group E (CG-E, CG1 in the U.S.A. and Europe), the PBDs of highest incidence, encoding the peroxin Pex1p of the AAA ATPase family. It has been also reported that Pex1p and Pex6p interact with each other. In the present study we investigated phenotype-genotype relationships of CG1 PBDs. Pex1p from IRD such as Pex1p with the most frequently identified mutation at G843D was largely degraded in vivo at 37 degrees C, whereas a normal level of Pex1p was detectable at the permissive temperature. In contrast, PEX1 proteins derived from ZS patients, including proteins with a mutation at L664P or the deletion of residues 634-690, were stably present at both temperatures. Pex1p-G843D interacted with Pex6p at approx. 50% of the level of normal Pex1p, whereas Pex1p from ZS patients mostly showing non-temperature-sensitive peroxisome biogenesis hardly bound to Pex6p. Taking these results together, we consider it most likely that the stability of Pex1p reflects temperature-sensitive peroxisome assembly in IRD fibroblasts. Failure in Pex1p-Pex6p interaction gives rise to more severe abnormalities, such as those manifested by patients with ZS. PMID- 11439092 TI - Targeting mutants of PTEN reveal distinct subsets of tumour suppressor functions. AB - The tumour suppressor protein PTEN (phosphatase and tensin homolog deleted on chromosome 10) is a lipid phosphatase which can antagonize the phosphoinositide 3 kinase (PI 3-kinase) signalling pathway, promoting apoptosis and inhibiting cell cycle progression and cell motility. We show that very little cellular PTEN is associated with the plasma membrane, but that artificial membrane-targeting of PTEN enhances its inhibition of signalling to protein kinase B (PKB). Evidence for potential targeting of PTEN to the membrane through PDZ domain-mediated protein-protein interactions led us to use a PTEN enzyme with a deletion of the C terminal PDZ-binding sequence, that retains full phosphatase activity against soluble substrates, and to analyse the efficiency of this mutant in different cellular assays. The extreme C-terminal PDZ-binding sequence was dispensable for the efficient down-regulation of cellular PtdIns(3,4,5)P3 levels and a number of PI 3-kinase-dependent signalling activities, including PKB and p70S6K. However, the PDZ-binding sequence was required for the efficient inhibition of cell spreading. The data show that a PTEN mutation, similar to those found in some tumours, affects some functions of the protein but not others, and implicate the deregulation of PTEN-dependent processes other than PKB activation in the development of some tumours. Significantly, this hypothesis is supported by data showing low levels of PKB phosphorylation in a glioblastoma sample carrying a mutation in the extreme C-terminus of PTEN compared with tumours carrying phosphatase-inactivating mutations of the enzyme. Our data show that deregulation of PKB is not a universal feature of tumours carrying PTEN mutations and implicate other processes that may be deregulated in these tumours. PMID- 11439093 TI - Phosphorylation of human plasma alpha2-Heremans-Schmid glycoprotein (human fetuin) in vivo. AB - A fraction of alpha2-Heremans-Schmid (alpha2-HS) glycoprotein (human fetuin) isolated from plasma was phosphorylated at serine-120 and serine-312 as shown by MS and peptide fragment sequencing after tryptic digestion. Serine-312-containing peptides were phosphorylated to 77% as determined from relative peak heights in the mass spectrum, which together with the phosphorylation of serine-120 implies a molar degree of phosphorylation of at least 1. Approximately 20% of the circulating fetuin plasma pool was phosphorylated to approx. 1 mol of phosphate/mol of protein. The remainder did not contain phosphate, resulting in an average phosphorylation degree for the protein in plasma of approx. 0.2 mol/mol. The isolated alpha2-HS glycoprotein was a heterodimer in which the entire C-terminal part of the connecting peptide including threonine-321 was present, but traces of C-terminally trimmed connecting peptide fragments were also found. The short B-chain was O-glycosylated to approx. 40%, whereas the N glycosylation of asparagine-138 and asparagine-158 seemed to be 100%. This finding, for the first time, that circulating human plasma fetuin is partly phosphorylated, implies that the effects of phosphorylated alpha2-HS glycoprotein on insulin signal transduction seen in different cell systems could be relevant to its physiological function in vivo. PMID- 11439094 TI - Cloning and characterization of the 5'-flanking region of the rat glutamate cysteine ligase catalytic subunit. AB - Glutamate-cysteine ligase (GCL), the rate-limiting enzyme in glutathione synthesis, is made up of two subunits, a catalytic (heavy) subunit (GCLC) and a modifier (light) subunit (GCLM), which are differentially regulated. Increased hepatic GCLC expression occurs during rapid growth, oxidative stress and after ethanol treatment. To facilitate studies of GCLC transcriptional regulation, we have cloned and characterized a 1.8 kb 5'-flanking region of the rat GCLC (GenBank accession number AF218362). A consensus TATA box and one transcriptional start site are located at 302 and 197 nucleotides upstream of the translational start site, respectively. The promoter contains consensus binding sites for many transcription factors including nuclear factor kappaB (NF-kappaB) and activator protein 1 (AP-1). The rat GCLC promoter was able to efficiently drive luciferase expression in H4IIE cells. Sequential deletion analysis revealed that three DNA regions, -595 to -111, -1108 to -705 and -705 to -595, are involved in positive (the first two regions) and negative (the latter region) gene regulation. Specific protein binding to these regions was confirmed by DNase I footprinting and electrophoretic mobility-shift assays (EMSAs). Ethanol-fed livers exhibit increased protein binding to region -416 to -336 on DNase I footprinting analysis, which was found to be NF-kappaB and AP-1 on EMSA and supershift analysis. Acetaldehyde treatment of H4IIE cells led to a time- and dose-dependent increase in GCLC mRNA levels, binding of NF-kappaB and AP-1 to the GCLC promoter, and luciferase activity driven by the GCLC promoter fragment containing these binding sites. PMID- 11439095 TI - Platelet-activating factor acetylhydrolase and transacetylase activities in human plasma low-density lipoprotein. AB - In this study, we demonstrate the presence of a transacetylase activity in human plasma low-density lipoprotein (LDL) that transfers short-chain fatty acids from platelet-activating factor (PAF) and its close ether- and ester-linked analogues to ether/ester-linked lysophospholipids (lyso-PL). We show evidence that both PAF acetylhydrolase (PAF-AH) and transacetylase activities are inhibited to the same extent by serine esterase inhibitors, are resistant to heat treatment, and exhibit identical distributions in lipoprotein classes and in LDL subfractions. Additionally, the competitive inhibition of PAF-AH by lyso-PL, and the evidence that the recombinant PAF-AH also showed a similar transacetylase activity, suggest that PAF-AH is responsible for both activities. Using PAF as a donor molecule and lyso-PAF (1-O-alkyl-sn-glycero-3-phosphocholine) as an acceptor, the transacetylase activity showed typical allosteric kinetics, due to the positive co-operativity of the substrates, with apparent Vmax=19.6+/-3.4 nmol/min per mg of protein, apparent h=2.0+/-0.3 and apparent [S]0.5=9.4+/-2.3 microM at saturation for the concentration of lyso-PAF. The substrate specificity of the donor molecules was decreased by increasing the chain length of the acyl moiety in the sn-2 position of the glycerol. The ether linkage in the sn-1 position of the substrate was 30% more effective than the ester bond; cholesteryl acetate was inactive as an acetyl donor. The two acceptors tested, lyso-PAF and the ester linked lyso-PC (1-acyl-sn-glycero-3-phosphocholine), showed similar specificity. Addition of exogenous lyso-PAF induced the transient formation of PAF-like aggregating activity predominantely in small dense LDL subfractions upon oxidation. We conclude that PAF-AH possesses both transacetylase and acetylhydrolase activities which remove PAF and its ether-linked analogues from LDL particles upon LDL oxidation. However, in atherogenic small dense LDL-5 particles, the transacetylase activity may acetylate extracellular lyso-PAF into biologically active PAF. PMID- 11439096 TI - FK506-binding protein of the hyperthermophilic archaeum, Thermococcus sp. KS-1, a cold-shock-inducible peptidyl-prolyl cis-trans isomerase with activities to trap and refold denatured proteins. AB - The FK506 (tacrolimus)-binding protein (FKBP) type peptidyl-prolyl cis-trans isomerase (PPIase) in the hyperthermophilic archaeum Thermococcus sp. KS-1 was shown to be induced by temperature downshift to growth temperatures lower than the optimum. This PPIase (TcFKBP18) showed chaperone-like protein refolding activity in addition to PPIase activity in vitro. It refolded unfolded citrate synthase (CS) and increased the yield of the refolded protein. At a molar ratio of 15:1 ([TcFKBP18] to [CS]) in the refolding mixture, the recovered yield of folded CS was maximal at 62%, whereas that of spontaneous refolding was 11%. Increasing FKBP above a 15:1 ratio decreased the final yield, whereas the aggregation of unfolded CS was suppressed. A cross-linking analysis showed the formation of a complex between TcFKBP18 and unfolded CS (1:1 complex) at molar ratios of 3:1 to 15:1. However, molar ratios of 15:1 or 60:1 induced the binding of multiple FKBP molecules to an unfolded CS molecule (multimeric complex). Disrupting hydrophobic interaction by adding ethylene glycol at a molar ratio of 60:1 ([TcFKBP18] to [CS]) suppressed the formation of this multimeric complex, simultaneously enhancing CS refolding. FK506 also suppressed the formation of the multimeric complex while increasing the chaperone-like activity. These results suggest that the hydrophobic region of TcFKBP18, probably the FK506-binding pocket, was important for the interaction with unfolded proteins. No cross-linked product was detected between TcFKBP18 and native dimeric CS. TcFKBP18 probably traps the unfolded protein, then refolds and releases it in a native form. This FKBP might be important at growth temperatures lower than the optimum in Thermococcus sp. KS-1 cells. PMID- 11439097 TI - Amphibian DNases I are characterized by a C-terminal end with a unique, cysteine rich stretch and by the insertion of a serine residue into the Ca2+-binding site. AB - We purified four amphibian deoxyribonucleases I from the pancreases of one toad, two frog and one newt species, by using three different column chromatography methods in sequence. Each of the purified enzymes had a molecular mass of approx. 40 kDa and an optimal pH for activity of approx. 8.0. These values were significantly greater than those for other vertebrate DNases I. The full-length cDNA encoding each amphibian DNase I was constructed from the total RNA of the pancreas by using rapid amplification of cDNA ends. Nucleotide sequence analyses revealed two structural characteristics unique to amphibian DNases I: a stretch of approx. 70 amino acids with a high cysteine content (approx. 15%) in the C terminal region, and the insertion of a serine residue at position 205 (in a domain containing an essential Ca2+-binding site). Expression analysis of a series of mutant constructs indicated that both of these structures are essential in generating the active form of the enzyme. 'DNase I signature sequences', which are well conserved in other vertebrate DNases I, could not be found in any of the amphibian DNases I tested, whereas a 'somatomedin B motif' was identified in the Cys-rich stretches of all four. Although DNase I has so far been considered to be a secretory glycoprotein, amphibian DNase I seems to be non-glycosylated. These structural findings indicate strongly that amphibian DNases I are situated in a unique position on the phylogenetic tree of the DNase I family. PMID- 11439098 TI - Comparison of the effects of dietary n-3 and n-6 polyunsaturated fatty acids on very-low-density lipoprotein secretion when delivered to hepatocytes in chylomicron remnants. AB - The effects of chylomicron remnants enriched in n-3 or n-6 polyunsaturated fatty acids (derived from fish or corn oil respectively) on the secretion of very-low density lipoprotein (VLDL) lipid and apolipoprotein B (apoB) by rat hepatocytes in culture was investigated. Remnants were prepared in vivo from chylomicrons obtained from rats given an oral dose of fish or corn oil and incubated with cultured hepatocytes for up to 16 h. The medium was then removed and the secretion of cholesterol and triacylglycerol into the whole medium or the rho<1.050 g/ml fraction during the following 7-24 h was determined. After exposure of the cells to fish-oil as compared with corn-oil remnants, secretion of both cholesterol and triacylglycerol into the whole medium was decreased by 25 35%, and secretion into the rho<1.050 g/ml fraction was decreased by 20-25%. In addition, the levels of apoB48 found in the rho<1.050 g/ml fraction were significantly lower in cells treated with fish-oil rather than corn-oil remnants, although the levels of apoB100 remained unchanged. The expression of mRNA for apoB, as determined by reverse-transcriptase PCR, however, was not significantly changed after exposure of the cells to both types of remnants. These results demonstrate that the effects of dietary n-3 polyunsaturated fatty acids in depressing hepatic VLDL secretion occur directly when they are delivered to the liver from the intestine in chylomicron remnants, and that the secretion, but not the synthesis, of apoB is targeted. PMID- 11439099 TI - Polyamines regulate gap junction communication in connexin 43-expressing cells. AB - The control of cell-cell communication through gap junctions is thought to be crucial in normal tissue function and during various stages of tumorigenesis. However, few natural regulators of gap junctions have been found. We show here that increasing the activity of ornithine decarboxylase, or adding polyamines to the outside of cells, increases the level of gap junction communication between various epithelial cells. Conversely, reduction of ornithine decarboxylase activity decreases the level of gap junction communication. This regulation is dependent upon the expression of connexin 43 (Cx43 or Cxalpha1), which is a major connexin expressed in many different cell types, and involves an increase in Cx43 and its cellular re-distribution. PMID- 11439100 TI - Wortmannin-sensitive trafficking steps in the endocytic pathway in rat liver endothelial cells. AB - Liver endothelial cells (LECs) play an important homoeostatic role by removing potentially harmful macromolecules from blood. The extremely efficient endocytosis in LECs makes these cells an interesting model for the study of the involvement of phosphoinositides in the different steps of the endocytic process. In the present investigation we have studied the effect of wortmannin, an inhibitor of phosphatidylinositol kinases, on uptake, recycling and intracellular transport of (125)I-labelled ovalbumin, which is taken up in LECs via mannose receptor-mediated endocytosis. Wortmannin was found to inhibit both uptake and degradation of ovalbumin. Further studies indicated that the reduced uptake via the mannose receptor was due both to a reduction of the number of surface receptors and a reduction in the rate of receptor-ligand internalization. Transport of ligand from endosomes to lysosomes was prevented, leading to increased recycling of internalized ligand. Wortmannin treatment released the Rab5 effector EEA1 from the endosomes and caused reduced size of early endosomes. PMID- 11439101 TI - Determination of the chemical structure of the capsular polysaccharide of strain B33, a fast-growing soya bean-nodulating bacterium isolated from an arid region of China. AB - We have determined the structure of a polysaccharide from strain B33, a fast growing bacterium that forms nitrogen-fixing nodules with Asiatic and American soya bean cultivars. On the basis of monosaccharide analysis, methylation analysis, one-dimensional 1H- and 13C-NMR and two-dimensional NMR experiments, the structure was shown to consist of a polymer having the repeating unit -->6)-4 O-methyl-alpha-D-Glcp-(1-->4)-3-O-methyl-beta-D-GlcpA-(1--> (where GlcpA is glucopyranuronic acid and Glcp is glucopyranose). Strain B33 produces a K-antigen polysaccharide repeating unit that does not have the structural motif sugar-Kdx [where Kdx is 3-deoxy-D-manno-2-octulosonic acid (Kdo) or a Kdo-related acid] proposed for different Sinorhizobium fredii strains, all of them being effective with Asiatic soya bean cultivars but unable to form nitrogen-fixing nodules with American soya bean cultivars. Instead, it resembles the K-antigen of S. fredii strain HH303 (rhamnose, galacturonic acid)n, which is also effective with both groups of soya bean cultivars. Only the capsular polysaccharide from strains B33 and HH303 have monosaccharide components that are also present in the surface polysaccharide of Bradyrhizobium elkanii strains, which consists of a 4-O-methyl D-glucurono-L-rhamnan. PMID- 11439102 TI - Involvement of the chicken liver 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase sequence His444-Arg-Glu-Arg in modulation of the bisphosphatase activity by its kinase domain. AB - The bisphosphatase activity of the hepatic bifunctional enzyme 6-phosphofructo-2 kinase/fructose-2,6-bisphosphatase is repressed by its kinase domain, and regulated by cAMP-dependent protein kinase (PKA)-catalysed phosphorylation. In the present study, the mechanism by which the bisphosphatase activity is repressed by the kinase domain and regulated by phosphorylation was investigated. We found that truncation of the C-terminus of the enzyme by 25, but not 20, amino acids dramatically enhanced the catalytic rate of the bisphosphatase, abrogated the inhibition by the kinase domain, and eliminated the effect of PKA-mediated phosphorylation on activity. In addition, mutation of His444-Arg-Glu-Arg to Ala Ala-Glu-Ala had similar effects as the deletion. Moreover, the mutations also significantly affected the phosphorylation-mediated regulation of the kinase activity of the enzyme. Furthermore, the mutations altered the pH-dependence of the bisphosphatase, and the mutant bisphosphatases were more sensitive to modification by diethyl pyrocarbonate and guanidine-induced inactivation than the wild-type enzyme. Taken together, these results demonstrate that the sequence His444-Arg-Glu-Arg plays a critical role in repression of the bisphosphatase activity by both the N-terminal kinase domain and the C-terminal tail itself. These results also explain the activation of the bisphosphatase activity by PKA catalysed phosphorylation, by suggesting that phosphorylation may relieve the inhibitory effect of the kinase domain that is mediated by the three basic residues in this sequence. PMID- 11439103 TI - Identification of a peroxisome-proliferator-activated-receptor response element in the apolipoprotein E gene control region. AB - Apolipoprotein E (apoE) is a protein involved in reverse cholesterol transport. Among other tissues, apoE is expressed in macrophages where its expression increases when macrophages develop into foam cells. It has been recently shown that peroxisome-proliferator-activated receptor gamma (PPARgamma) is involved in this conversion. Northern-blot analysis was carried out in the macrophage cell line THP1 to determine whether apoE mRNA levels were regulated by ciglitazone, a PPARgamma inducer. The results indicated that treatment with ciglitazone doubled the levels of apoE mRNA. To identify a possible PPARgamma response element (PPRE), several portions of apoE gene control region were used to construct luciferase reporter plasmids. In U-87 MG cells, a 185 bp fragment located in the apoE/apoCI intergenic region was sufficient to induce a 10-fold increase in the luciferase activity of the extract of cells co-transfected with a PPARgamma expression plasmid. Subsequent analysis revealed the presence of a sequence with a high level of sequence similarity to the consensus PPRE. Mutations in this sequence resulted in a lack of functionality both in transient transfection and in electrophoretic-mobility-shift assays. These results demonstrated the presence of a functional PPRE in the apoE/apoCI intergenic region. These results have implications for the regulation of apoE gene expression and could be relevant for understanding the anti-atherogenic effect of thiazolidinediones. PMID- 11439104 TI - Green fluorescent protein (GFP) tagged to the cytoplasmic tail of alphaIIb or beta3 allows the expression of a fully functional integrin alphaIIb(beta3): effect of beta3GFP on alphaIIb(beta3) ligand binding. AB - Using green fluorescent protein (GFP) as an autofluorescent tag, we report the first successful visualization of a beta3 integrin in a living cell. GFP fused in frame to the cytoplasmic tail of either alphaIIb or beta3 allowed normal expression, heterodimerization, processing and surface exposure of alphaIIbGFPbeta3 and alphaIIb(beta3)GFP receptors in Chinese hamster ovary (CHO) cells. Direct microscopic observation of the autofluorescent cells in suspension following antibody-induced alphaIIb(beta3) capping revealed an intense autofluorescent cap corresponding to unlabelled immunoclustered GFP-tagged alphaIIb(beta3). GFP-tagged alphaIIbbeta3 receptors mediated fibrinogen-dependent cell adhesion, were readily detectable in focal adhesions of unstained living cells and triggered p125(FAK) tyrosine phosphorylation similar to wild-type alphaIIb(beta3) (where FAK corresponds to focal adhesion kinase). However, GFP tagged to beta3, but not to alphaIIb, induced spontaneous CHO cell aggregation in the presence of soluble fibrinogen, as well as binding of the fibrinogen mimetic monoclonal antibody PAC1 in the absence of alphaIIb(beta3) receptor activation. Time-lapse imaging of living transfectants revealed a characteristic redistribution of GFP-tagged alphaIIb(beta3) during the early stages of cell attachment and spreading, starting with alphaIIb(beta3) clustering at the rim of the cell contact area, that gradually overlapped with the boundary of the attached cell, and, with the onset of cell spreading, to a reorganization of alphaIIb(beta3) in focal adhesions. Taken together, our results demonstrate that (1) fusion of GFP to the cytoplasmic tail of either alphaIIb or beta3 integrin subunits allows normal cell surface expression of a functional receptor, and (2) structural modification of the beta3 integrin cytoplasmic tail, rather than the alphaIIb subunit, plays a major role in alphaIIb(beta3) affinity modulation. With the successful direct visualization of functional alphaIIb(beta3) receptors in living cells, the generation of autofluorescent integrins in transgenic animals will become possible, allowing new approaches to study the dynamics of integrin functions. PMID- 11439105 TI - Protein-binding elements in the proximal parotid secretory protein gene enhancer essential for salivary-gland-specific expression. AB - The murine parotid secretory protein (PSP) gene is expressed at high levels in the parotid gland and at lower levels in the sublingual gland. A proximal enhancer core necessary for tissue-specific expression was identified previously, and it was demonstrated that one element, parotid gland element I (PGE I), exhibited specific binding of parotid gland nuclear proteins. In the present study, we demonstrate that a related adjacent element, PGE II, which binds nuclear proteins in a much less tissue-restricted manner, is able to compete with PGE I for binding of parotid-gland-specific factors. The functional significance of PGE I and PGE II was examined in transgenic mice. Deletion of PGE II reduced transgene expression only in the parotid gland, whereas deletion of PGE I appeared to reduce expression in both of the PSP-expressing salivary glands. Combined deletion of PGE I and PGE II reduced expression below the limit of detection. Thus PGE I and PGE II are functionally important salivary-gland specific binding elements that are necessary for the salivary-gland-specific expression of a PSP minigene in transgenic mice. PMID- 11439106 TI - Functional and conformational properties of the exclusive C-domain from the Arabidopsis copper chaperone (CCH). AB - The Arabidopsis thaliana copper chaperone (CCH) is a small copper binding protein involved in copper trafficking. When compared to homologues from other eukaryotes, CCH has two different domains; the conserved N-domain and the plant exclusive C-domain, a C-terminal extension with an unusual amino-acid composition. In order to characterize this extra C-domain, the CCH protein, the N domain and the C-domain were all expressed separately in heterologous systems. While the N-domain retained the copper chaperone and antioxidant properties described for the yeast Atx1 and human HAH1 counterparts, the C-domain displayed particular structural properties that would be necessary to optimize copper homoeostasis in plant cells where it could be responsible for the metallochaperone plant-exclusive intercellular transport. The whole CCH protein and the C-domain, but not the N-domain, displayed altered SDS/PAGE mobilities. CD spectroscopy showed that the N-domain fold is representative of an alpha/beta protein, while the C-domain adopts an extended conformation. PMID- 11439107 TI - Human cyritestin genes (CYRN1 and CYRN2) are non-functional. AB - The mouse cyritestin gene is a member of the ADAM (a disintegrin and metalloprotease) gene family and codes for a membrane-anchored sperm protein. Recently, it was shown that cyritestin is critical for male fertility in the mouse. Spermatozoa of cyritestin-deficient mice are not able to bind to the zona pellucida of the oocyte and therefore unable to fertilize the egg. However, zona free oocytes can be fertilized and the resulting embryos show normal development. In contrast to the mouse, where only one gene for cyritestin (Cyrn) is reported, two cyritestin genes (CYRN1 and CYRN2) are known in humans. The human CYRN1 and CYRN2 genes are located on chromosomes 8 and 16, respectively. We report that 27% of fertile men are deficient for the CYRN1 gene but that all have a CYRN2 gene, suggesting that the CYRN2 gene is the orthologous mouse cyritestin gene in humans and might be involved in sperm-egg interactions. However, the characterization of CYRN2 transcripts from testicular RNA of CYRN1-deficient men demonstrated many termination codons in the synthesized cyritestin cDNA. Furthermore, Western-blot analysis with human testicular protein extracts using an anti-cyritestin antibody failed to detect any cyritestin protein. These results demonstrate clearly that both cyritestin genes are non-functional in humans. PMID- 11439108 TI - Glucose 6-phosphate transport in fibroblast microsomes from glycogen storage disease type 1b patients: evidence for multiple glucose 6-phosphate transport systems. AB - In liver endoplasmic reticulum the intralumenal glucose-6-phosphatase activity requires the operation of a glucose 6-phosphate transporter (G6PT1). Mutations in the gene encoding G6PT1 cause glycogen storage disease type 1b, which is characterized by a loss of glucose-6-phosphatase activity and impaired glucose homoeostasis. We describe a novel glucose 6-phosphate (G6P) transport activity in microsomes from human fibroblasts and HeLa cells. This transport activity is unrelated to G6PT1 since: (i) it was similar in microsomes of skin fibroblasts from glycogen storage disease type 1b patients homozygous for mutations of the G6PT1 gene, and in microsomes from human control subjects; (ii) it was insensitive to the G6PT1 inhibitor chlorogenic acid; and (iii) it was equally active towards G6P and glucose 1-phosphate, whereas G6PT1 is highly selective for G6P. Taken together, our results provide evidence for the presence of multiple transporters for G6P (and other hexose phosphoesters) in the endoplasmic reticulum. PMID- 11439109 TI - Autocatalytic tyrosine-phosphorylation of protein kinase CK2 alpha and alpha' subunits: implication of Tyr182. AB - CK2 is a pleiotropic and constitutively active serine/threonine protein kinase composed of two catalytic (alpha and/or alpha') and two regulatory beta-subunits, whose mechanism of modulation is still obscure. Here we show that CK2 alpha/alpha' subunits undergo intermolecular (trans) tyrosine autophosphorylation, which is dependent on intrinsic catalytic activity and is suppressed by the individual mutation of Tyr182, a crucial residue of the activation loop, to phenylalanine. At variance with serine-autophosphorylation, tyrosine-autophosphorylation of CK2alpha is reversed by ADP and GDP and is counteracted by the beta-subunit and by a peptide reproducing the activation loop of CK2alpha/alpha' (amino acids 175-201). These results disclose new perspectives about the mode of regulation of CK2 catalytic subunits. PMID- 11439110 TI - Extracellular processing of amphoterin generates a peptide active on erythroleukaemia cell differentiation. AB - The release of amphoterin by murine erythroleukaemia cells exposed to the chemical inducer hexamethylenebisacetamide represents an essential step for the process of their terminal differentiation. Once exported in the culture medium, amphoterin undergoes limited proteolysis, catalysed by a serine proteinase also secreted by stimulated cells. The isolated proteinase is responsible for degradation of amphoterin, with the production of a 10-amino-acid-residue fragment, specifically retaining the cell-differentiation-stimulating activity of the native protein molecule. This peptide does not express other properties of amphoterin, such as protein kinase C-stimulating activity or systemic toxicity. These findings define a selective mechanism accounting for extracellular amphoterin functional maturation. PMID- 11439111 TI - Inhibition of distant caspase homologues by natural caspase inhibitors. AB - Caspases play an important role in the ability of animal cells to kill themselves by apoptosis. Caspase activity is regulated in vivo by members of three distinct protease inhibitor families, two of which, baculovirus p35 and members of the inhibitor of apoptosis (IAP) family, are thought to be caspase specific. However, caspases are members of the clan of cysteine proteases designated CD, which also includes animal and plant legumains, and the bacterial proteases clostripain, gingipain-R and gingipain-K. Since these proteases have been proposed to have a common mechanism and evolutionary origin, we hypothesized that the caspase inhibitors may also regulate these other proteases. We tested this hypothesis by examining the effect of the natural caspase inhibitors on other members of protease clan CD. The IAP family proteins were found to have only a slight inhibitory effect on gingipain-R. The cowpox viral cytokine-response modifier A (CrmA) serpin had no effect on any of the proteases tested but a single point mutation of CrmA (Asp-->Lys) resulted in strong inhibition of gingipain-K. More substantial, with respect to the hypothesis, was the strong inhibition of gingipain-K by wild-type p35. The site in p35, required for inhibition of gingipain-K, was mapped to Lys94, seven residues C-terminal to the caspase inhibitory site. Our data indicate that the virally encoded caspase inhibitors have adopted a mechanism that allows them to regulate disparate members of clan CD proteases. PMID- 11439112 TI - Sulphoacetaldehyde sulpho-lyase (EC 4.4.1.12) from Desulfonispora thiosulfatigenes: purification, properties and primary sequence. AB - The strictly anaerobic bacterium Desulfonispora thiosulfatigenes ferments taurine via sulphoacetaldehyde, which is hydrolysed to acetate and sulphite by sulphoacetaldehyde sulpho-lyase (EC 4.4.1.12). The lyase was expressed at high levels and a two-step, 4.5-fold purification yielded an apparently homogeneous soluble protein, which was presumably a homodimer in its native form; the molecular mass of the subunit was about 61 kDa (by SDS/PAGE). The mass was determined to be 63.8 kDa by matrix-assisted laser-desorption ionization-time-of flight (MALDI-TOF) MS. The purified enzyme converted 1 mol of sulphoacetaldehyde to 1 mol each of sulphite and acetate, but no requirement for thiamine pyrophosphate (TPP) was detected. The N-terminal and two internal amino acid sequences were determined, which allowed us to generate PCR primers. The gene was amplified and sequenced. The DNA sequence had no significant homologue in the databases searched, whereas the derived amino acid sequence indicated an oxo-acid lyase, revealed a TPP-binding site and gave a derived molecular mass of 63.8 kDa. PMID- 11439113 TI - G-protein-coupled-receptor activation of the smooth muscle calponin gene. AB - A hallmark of cultured smooth muscle cells (SMCs) is the rapid down-regulation of several lineage-restricted genes that define their in vivo differentiated phenotype. Identifying factors that maintain an SMC differentiated phenotype has important implications in understanding the molecular underpinnings governing SMC differentiation and their subversion to an altered phenotype in various disease settings. Here, we show that several G-protein coupled receptors [alpha-thrombin, lysophosphatidic acid and angiotensin II (AII)] increase the expression of smooth muscle calponin (SM-Calp) in rat and human SMC. The increase in SM-Calp protein appears to be selective for G-protein-coupled receptors as epidermal growth factor was without effect. Studies using AII showed a 30-fold increase in SM-Calp protein, which was dose- and time-dependent and mediated by the angiotensin receptor-1 (AT1 receptor). The increase in SM-Calp protein with AII was attributable to transcriptional activation of SM-Calp based on increases in steady-state SM-Calp mRNA, increases in SM-Calp promoter activity and complete abrogation of protein induction with actinomycin D. To examine the potential role of extracellular signal-regulated kinase (Erk1/2), protein kinase B, p38 mitogen activated protein kinase and protein kinase C in AII-induced SM-Calp, inhibitors to each of the signalling pathways were used. None of these signalling molecules appears to be crucial for AII-induced SM-Calp expression, although Erk1/2 may be partially involved. These results identify SM-Calp as a target of AII-mediated signalling, and suggest that the SMC response to AII may incorporate a novel activity of SM-Calp. PMID- 11439114 TI - The shoot apical meristem restores its symplasmic organization during chilling induced release from dormancy. AB - The shoot apex of overwintering perennials ceases its morphogenetic activity at the end of the growing season and transforms into a bud which is dormant and freezing-tolerant. In birch (Betula pubescens) these events are triggered by short photoperiod, and involve the production of 1,3-beta-D-glucan containing sphincters on the plasmodesmata. As a result, all symplasmic pathways shut down. Here we show that breakage of bud dormancy by chilling involves restoration of the symplasmic organization of the meristem. This restoration is likely to be mediated by 1,3-beta-D-glucanase, which was present in small spherosome-like vacuoles that arose de novo during dormancy induction. During chilling these vacuoles were displaced from the bulk cytoplasm to the cortical cytoplasm where they became aligned with the plasma membrane, often associated with plasmodesmata. At this stage the enzyme also appeared outside the vacuoles. During chilling, 1,3-beta-D-glucan disappeared from the plasmodesmal channels and wall sleeves, and the plasmodesmata regained the capacity for cell-cell transport, as demonstrated by microinjection of Lucifer Yellow CH and Fluorescein tagged gibberellic acid. Collectively, the present experiments demonstrate that restoration of the symplasmic organization of the meristem is indispensable for the release of buds from dormancy and the assumption of a proliferation-competent state, and implicate 1,3-beta-D-glucanase action at the plasmodesmata. Based on these findings we propose a model for 'dormancy cycling' which depicts the meristem as passing through three sequential states of cellular communication with characteristic sensitivities to distinct environmental cues. PMID- 11439115 TI - Dissection of RNA-mediated ryegrass mosaic virus resistance in fertile transgenic perennial ryegrass (Lolium perenne L.). AB - Ryegrass mosaic virus (RgMV) frequently reduces yield and persistence of perennial ryegrass. We introduced an untranslatable RgMV coat protein (RgMV-CP) gene into perennial ryegrass using particle bombardment to explore the potential of RNA-mediated virus resistance. Nine months after inoculation with the RgMV Bulgaria strain, the most resistant transgenic line showed no immuno-detectable RgMV-CP in all replications of the primary transformant and a sexual progeny. A significant influence of the transgenic line, virus strain and the period after inoculation on the RgMV level was observed. Typical examples of recovery resistance from initially susceptible plants were not identified. Molecular analysis revealed RgMV resistance operates by targeted RNA degradation resulting in post-transcriptional transgene silencing (PTGS) along with inhibition of virus RNA replication and was not associated with a threshold of transgene expression. Suppression of PTGS by RgMV differed significantly in two progenies with identical RgMV-CP transgene integration and a similar RgMV-CP transgene methylation pattern, suggesting the presence of quantitative components controlling the stability of PTGS in perennial ryegrass. This study extends RNA mediated virus resistance, post transcriptional gene silencing and crop improvement by genetic engineering to an agronomically important perennial monocot. PMID- 11439116 TI - Reconstruction of reproductive diversity in Hypericum perforatum L. opens novel strategies to manage apomixis. AB - The mode of reproduction was characterized for 113 accessions of the tetraploid facultative apomictic species Hypericum perforatum using bulked or single mature seeds in the flow cytometric seed screen (FCSS). This screen discriminates several processes of sexual or asexual reproduction based on DNA contents of embryo and endosperm nuclei. Seed formation in H. perforatum proved to be highly polymorphic. Eleven different routes of reproduction were determined. For the first time, individual seeds were identified that originated from two embryo sacs: the endosperm from an aposporous and the embryo from the legitimate meiotic embryo sac. Moreover, diploid plants were discovered, which apparently reproduce by a hitherto unknown route of seed formation, that is chromosome doubling within aposporous initial cells followed by double fertilization. Although most plants were tetraploid and facultative sexual/apomictic, diploid obligate sexuals and tetraploid obligate apomicts could be selected. Additionally, genotypes were detected which at a high frequency produced embryos either from reduced parthenogenetic or unreduced fertilized egg cells. The endosperm developed most frequently after fertilization of the central cell in aposporous embryo sacs (pseudogamy) but in few cases also autonomously. The genetic control of apomixis appears to be complex in H. perforatum. Basic material was developed for breeding H. perforatum, and strategies are suggested for elucidation of inheritance as well as evolution of apomixis and for molecular approaches of apomixis engineering. PMID- 11439117 TI - Interaction of phytochromes A and B in the control of de-etiolation and flowering in pea. AB - The interactions of phytochrome A (phyA) and phytochrome B (phyB) in the photocontrol of vegetative and reproductive development in pea have been investigated using null mutants for each phytochrome. White-light-grown phyA phyB double mutant plants show severely impaired de-etiolation both at the seedling stage and later in development, with a reduced rate of leaf production and swollen, twisted internodes, and enlarged cells in all stem tissues. PhyA and phyB act in a highly redundant manner to control de-etiolation under continuous, high-irradiance red light. The phyA phyB double mutant shows no significant residual phytochrome responses for either de-etiolation or shade-avoidance, but undergoes partial de-etiolation in blue light. PhyB is shown to inhibit flowering under both long and short photoperiods and this inhibition is required for expression of the promotive effect of phyA. PhyA is solely responsible for the promotion of flowering by night-breaks with white light, whereas phyB appears to play a major role in detection of light quality in end-of-day light treatments, night breaks and day extensions. Finally, the inhibitory effect of phyB is not graft-transmissible, suggesting that phyB acts in a different manner and after phyA in the control of flower induction. PMID- 11439118 TI - Analysis of the SINE S1 Pol III promoter from Brassica; impact of methylation and influence of external sequences. AB - Transcription is an important control point in the transposable element mobilization process. To better understand the regulation of the plant SINE (Short Interspersed Elements) S1, its promoter sequence was studied using an in vitro pol III transcription system derived from tobacco cells. We show that the internal S1 promoter can be functional although upstream external sequences were found to enhance this basal level of transcription. For one putative 'master' locus (na7), three CAA triplets (in positions -12, -7 and -2) and two overlapping TATA motifs (in positions -54 to -43) were important to stimulate transcription. For this locus, two transcription initiation regions were characterized, one centered on position + 1 (first nucleotide of the S1 element) and one centered on position - 19 independently of the internal motifs. The CAA triplets only influence transcription in + 1 and work in association with the internal motifs. We show that methylation can inhibit transcription at the na7 locus. We also observe that S1 RNA is cleaved in a smaller Poly (A) minus product by a process analogous to the maturation of mammalian SINEs. PMID- 11439119 TI - Analysis of a contiguous 211 kb sequence in diploid wheat (Triticum monococcum L.) reveals multiple mechanisms of genome evolution. AB - In plant species with large genomes such as wheat or barley, genome organization at the level of DNA sequence is largely unknown. The largest sequences that are publicly accessible so far from Triticeae genomes are two 60 kb and 66 kb intervals from barley. Here, we report on the analysis of a 211 kb contiguous DNA sequence from diploid wheat (Triticum monococcum L.). Five putative genes were identified, two of which show similarity to disease resistance genes. Three of the five genes are clustered in a 31 kb gene-enriched island while the two others are separated from the cluster and from each other by large stretches of repetitive DNA. About 70% of the contig is comprised of several classes of transposable elements. Ten different types of retrotransposons were identified, most of them forming a pattern of nested insertions similar to those found in maize and barley. Evidence was found for major deletion, insertion and duplication events within the analysed region, suggesting multiple mechanisms of genome evolution in addition to retrotransposon amplification. Seven types of foldback transposons, an element class previously not described for wheat genomes, were characterized. One such element was found to be closely associated with genes in several Triticeae species and may therefore be of use for the identification of gene-rich regions in these species. PMID- 11439120 TI - Coregulation of light-harvesting complex II phosphorylation and lhcb mRNA accumulation in winter rye. AB - Winter rye plants grown under contrasting environmental conditions or just transiently shifted to varying light and temperature conditions, were studied to elucidate the chloroplast signal involved in regulation of photosynthesis genes in the nucleus. Photosystem II excitation pressure, reflecting the plastoquinone redox state, and the phosphorylation level of thylakoid light-harvesting proteins (LHCII and CP29) were monitored together with changes occurring in the accumulation of lhcb, rbcS and psbA mRNAs. Short-term shifts of plants to changed conditions, from 1 h to 2 d, were postulated to reveal signals crucial for the initiation of the acclimation process. Comparison of these results with those obtained from plants acclimated during several weeks' growth at contrasting temperature and in different light regimes, allow us to make the following conclusions: (1) LHCII protein phosphoylation is a sensitive tool to monitor redox changes in chloroplasts; (2) LHCII protein phosphorylation and lhcb mRNA accumulation occur under similar conditions and are possibly coregulated via an activation state of the same kinase (the LHCII kinase); (3) Maximal accumulation of lhcb mRNA during the diurnal light phase seems to require an active LHCII kinase whereas inactivation of the kinase is accompanied by dampening of the circadian oscillation in the amount of lhcb mRNA; (4) Excitation pressure of photosystem II (reduction state of the plastoquinone pool) is not directly involved in the regulation of lhcb mRNA accumulation. Instead (5) the redox status of the electron acceptors of photosystem I in the stromal compartment seems to be highly regulated and crucial for the regulation of lhcb gene expression in the nucleus. PMID- 11439121 TI - Arabidopsis UVH3 gene is a homolog of the Saccharomyces cerevisiae RAD2 and human XPG DNA repair genes. AB - To identify mechanisms of DNA repair in Arabidopsis thaliana, we have analyzed a mutant (uvh3) which exhibits increased sensitivity to ultraviolet (UV) light, H2O2 and ionizing radiation and displays a premature senescence phenotype. The uvh3 locus was mapped within chromosome III to the GL1 locus. A cosmid contig of the GL1 region was constructed, and individual cosmids were used to transform uvh3 mutant plants. Cosmid N9 was found to confer UV-resistance, H2O2-resistance and a normal senescence phenotype following transformation, indicating that the UVH3 gene is located on this cosmid and that all three phenotypes are due to the same mutation. Analysis of cosmid N9 sequences identified a gene showing strong similarity to two homologous repair genes, RAD2 (Saccharomyces cerevisiae) and XPG (human), which encode an endonuclease required for nucleotide excision repair of UV-damage. The uvh3 mutant was shown to carry a nonsense mutation in the coding region of the AtRAD2/XPG gene, thus revealing that the UVH3 gene encodes the AtRAD2/XPG gene product. In humans, the homologous XPG protein is also involved in removal of oxygen-damaged nucleotides by base excision repair. We discuss the possibility that the increased sensitivity of the uvh3 mutant to H2O2 and the premature senescence phenotype might result from failure to repair oxygen damage in plant tissues. Finally, we show that the AtRAD2/XPG gene is expressed at moderate levels in all plant tissues. PMID- 11439122 TI - Collateral gene expression changes induced by distinct plant viruses during the hypersensitive resistance reaction in Chenopodium amaranticolor. AB - Hypersensitive reactions to plant diseases are typically mediated by R genes. Many R genes that have been cloned only confer resistance to a particular pathogen. However, Chenopodium spp. have multivirus hypersensitive resistance, thus making the understanding of this broad-spectrum resistance mechanism attractive. Using tobacco mosaic virus (TMV) tagged with green fluorescent protein to follow infection over time, cDNA-AFLP to find genes up-regulated during virus infection in C. amaranticolor and quantitative RT-PCR to accurately measure gene expression at different time points, the first dissection of this significant defense response pathway is presented. The detected disease-expressed sequences in C. amaranticolor (DESCA) are similar to those that encode p450 monooxegenases, hypersensitivity-related genes, cellulases, ABC transporters, receptor-like kinases, serine/threonine kinases, phosphoribosylanthranilate transferases and hypothetical R genes, many of which are associated with pathogen defense in other plants. The expressions of these DESCA genes are also induced by infection with the taxonomically distinct tobacco rattle virus (TRV) in C. amaranticolor. In particular, DESCA1, one of the gene fragments from C. amaranticolor that lacks similarity to any other sequence in the GenBank database, is induced at least 200 fold 4 d after infection (dai) by both TMV and TRV. The potential role of DESCA genes in a C. amaranticolor multivirus defense response with regard to their levels and time of gene expression is discussed. PMID- 11439123 TI - The role of calcium in ABA-induced gene expression and stomatal movements. AB - There is much interest in the transduction pathways by which abscisic acid (ABA) regulates stomatal movements (ABA-turgor signalling) and by which this phytohormone regulates the pattern of gene expression in plant cells (ABA-nuclear signalling). A number of second messengers have been identified in both the ABA turgor and ABA-nuclear signalling pathways. A major challenge is to understand the architecture of ABA-signalling pathways and to determine how the ABA signal is coupled to the appropriate response. We have investigated whether separate Ca2+-dependent and -independent ABA-signalling pathways are present in guard cells. Our data suggest that increases in [Ca2+]i are a common component of the guard cell ABA-turgor and ABA-nuclear signalling pathways. The effects of Ca2+ antagonists on ABA-induced stomatal closure and the ABA-responsive CDeT6-19 gene promoter suggest that Ca2+ is involved in both ABA-turgor signalling and ABA nuclear signalling in guard cells. However, the sensitivity of these pathways to alterations in the external calcium concentration differ, suggesting that the ABA nuclear and ABA-turgor signalling pathways are not completely convergent. Our data suggest that whilst Ca2+-independent signalling elements are present in the guard cell, they do not form a completely separate Ca2+-independent ABA signalling pathway. PMID- 11439124 TI - Overexpression of chlorophyllide a oxygenase (CAO) enlarges the antenna size of photosystem II in Arabidopsis thaliana. AB - The light-harvesting efficiency of a photosystem is thought to be largely dependent on its photosynthetic antenna size. It has been suggested that antenna size is controlled by the biosynthesis of chlorophyll b. To verify this hypothesis, we overexpressed the enzyme for chlorophyll b biosynthesis, chlorophyllide a oxygenase (CAO), in Arabidopsis thaliana by transforming the plant with cDNA for CAO under the control of the 35S cauliflower mosaic virus promoter. In the early de-etiolation phase, when the intrinsic CAO expression is very low, the chlorophyll a: b ratio was drastically decreased from 28 to 7.3, indicating that enhancement of chlorophyll b biosynthesis had been successfully achieved. We made the following observations in full-green rosette leaves of transgenic plants. (1) The chlorophyll a : b ratio was reduced from 2.85 to 2.65. (2) The ratio of the peripheral light-harvesting complexes (LHCII) to the core antenna complex (CPa) resolved with the green-gel system increased by 20%. (3) The ratio of the light-harvesting complex II apoproteins (LHCP) to 47-kDa chlorophyll a protein (CP47), which was estimated by the results of immunoblotting, increased by 40%. These results indicated that the antenna size increased by at least 10-20% in transgenic plants, suggesting that chlorophyll b biosynthesis controls antenna size. To the best of our knowledge, this is the first report on enlargement of the antenna size by genetic manipulations. PMID- 11439125 TI - Presence of an SAR-like sequence in junction regions between an introduced transgene and genomic DNA of cultured tobacco cells: its effect on transformation frequency. AB - A 12.5-kb DNA fragment with junction regions between the transgene and genomic DNA was cloned from a transgenic tobacco cell line obtained by microprojectile bombardment of plasmid pCaMVNEO. Nucleotide sequence analysis of the fragment (DDBJ accession no. D84238) showed that it carried a 7.7-kb core sequence (concatemer of a complete pCaMVNEO and a partial pCaMVNEO) and two identical 1.3 kb junction sequences that flanked both the 5' and 3' ends of the core sequence and had inverted orientations. These sequences had topoisomerase II (Topo II) cleavage sites and adenine and thimine-rich sequences known to be specific to nuclear scaffold-attachment regions (SARs). An in vitro binding assay showed that a 507-bp fragment (designated TJ1) from the 1.3-kb sequence had the ability to bind to nuclear scaffold preparations of cultured tobacco cells, confirmation that the 1.3-kb sequence is an SAR. Insertion of TJ1 at the 5' and 3' sides of the expression cassette for the npt II gene increased transformant yields 5- to 10-fold and the NPT II enzyme activity per copy of the gene 5-fold. TJ1 enhances the integration or expression of the transgene, or both. Clearly, TJ1 is very useful for producing transgenic plants. This is the first report on an SAR-like sequence that is located in the transgene locus and enhances transformation efficiency in eukaryotic cells. The possible role of TJ1-SAR in the molecular evolution of plant genome is discussed. PMID- 11439126 TI - APETALA1 and SEPALLATA3 interact to promote flower development. AB - In Arabidopsis, the closely related APETALA1 (AP1) and CAULIFLOWER (CAL) MADS-box genes share overlapping roles in promoting flower meristem identity. Later in flower development, the AP1 gene is required for normal development of sepals and petals. Studies of MADS-domain proteins in diverse species have shown that they often function as heterodimers or in larger ternary complexes, suggesting that additional proteins may interact with AP1 and CAL during flower development. To identify proteins that may interact with AP1 and CAL, we used the yeast two hybrid assay. Among the five MADS-box genes identified in this screen, the SEPALLATA3 (SEP3) gene was chosen for further study. Mutations in the SEP3 gene, as well as SEP3 antisense plants that have a reduction in SEP3 RNA, display phenotypes that closely resemble intermediate alleles of AP1. Furthermore, the early flowering phenotype of plants constitutively expressing AP1 is significantly enhanced by constitutive SEP3 expression. Taken together, these studies suggest that SEP3 interacts with AP1 to promote normal flower development. PMID- 11439127 TI - Constitutive salicylic acid-dependent signaling in cpr1 and cpr6 mutants requires PAD4. AB - Salicylic acid (SA)-dependent signaling controls activation of a set of plant defense mechanisms that are important for resistance to a variety of microbial pathogens. Many Arabidopsis mutants that display altered SA-dependent signaling have been isolated. We used double mutant analysis to determine the relative positions of the pad4, cpr1, cpr5, cpr6, dnd1 and dnd2 mutations in the signal transduction network leading to SA-dependent activation of defense gene expression and disease resistance. The pad4 mutation causes failure of SA accumulation in response to infection by certain pathogens, while the other mutations cause constitutively high levels of SA, defense gene expression and resistance. The cpr1 pad4, cpr5 pad4, cpr6 pad4, dnd1 pad4 and dnd2 pad4 double mutants were constructed and assayed for stature, presence of spontaneous lesions, resistance to Pseudomonas syringae and Peronospora parasitica, SA levels, expression of PAD4, PR-1 and PDF1.2, and accumulation of camalexin. We found that the effects of the cpr1 and cpr6 mutations on SA-dependent gene expression are completely dependent on PAD4 function. In contrast, SA accumulation in the lesion-mimic mutant cpr5 is partially PAD4-independent, while in dnd1 and dnd2 mutants it is completely PAD4-independent. A model describing a possible arrangement of activities in the signal transduction network is presented. PMID- 11439128 TI - Constitutive disease resistance requires EDS1 in the Arabidopsis mutants cpr1 and cpr6 and is partially EDS1-dependent in cpr5. AB - The systemic acquired resistance (SAR) response in Arabidopsis is characterized by the accumulation of salicylic acid (SA), expression of the pathogenesis related (PR) genes, and enhanced resistance to virulent bacterial and oomycete pathogens. The cpr (constitutive expressor of PR genes) mutants express all three SAR phenotypes. In addition, cpr5 and cpr6 induce expression of PDF1.2, a defense related gene associated with activation of the jasmonate/ethylene-mediated resistance pathways. cpr5 also forms spontaneous lesions. In contrast, the eds1 (enhanced disease susceptibility) mutation abolishes race-specific resistance conferred by a major subclass of resistance (R) gene products in response to avirulent pathogens. eds1 plants also exhibit increased susceptibility to virulent pathogens. Epistasis experiments were designed to explore the relationship between the cpr- and EDS1-mediated resistance pathways. We found that a null eds1 mutation suppresses the disease resistance phenotypes of both cpr1 and cpr6. In contrast, eds1 only partially suppresses resistance in cpr5, leading us to conclude that cpr5 expresses both EDS1-dependent and EDS1 independent components of plant disease resistance. Although eds1 does not prevent lesion formation on cpr5 leaves, it alters their appearance and reduces their spread. This phenotypic difference is associated with increased pathogen colonization of cpr5 eds1 plants compared to cpr5. The data allow us to place EDS1 as a necessary downstream component of cpr1- and cpr6-mediated responses, but suggest a more complex relationship between EDS1 and cpr5 in plant defense. PMID- 11439129 TI - Impaired sucrose-induction mutants reveal the modulation of sugar-induced starch biosynthetic gene expression by abscisic acid signalling. AB - Plants both produce and utilize carbohydrates and have developed mechanisms to regulate their sugar status and co-ordinate carbohydrate partitioning. High sugar levels result in a feedback inhibition of photosynthesis and an induction of storage processes. We used a genetic approach to isolate components of the signalling pathway regulating the induction of starch biosynthesis. The regulatory sequences of the sugar inducible ADP-glucose pyrophosphorylase subunit ApL3 were fused to a negative selection marker. Of the four impaired sucrose induction (isi) mutants described here, two (isi1 and isi2) were specific to this screen. The other two mutants (isi3 and isi4) showed additional phenotypes associated with sugar-sensing screens that select for seedling establishment on high-sugar media. The isi3 and isi4 mutants were found to be involved in the abscisic acid signalling pathway. isi3 is allelic to abscisic acid insensitive4 (abi4), a gene encoding an Apetala2-type transcription factor; isi4 was found to be allelic to glucose insensitive1 (gin1) previously reported to reveal cross talk between ethylene and glucose signalling. Here we present an alternative interpretation of gin1 as an allele of the ABA-deficient mutant aba2. Expression analysis showed that ABA is unable to induce ApL3 gene expression by itself, but greatly enhances ApL3 induction by sugar. Our data suggest a major role for ABA in relation to sugar-signalling pathways, in that it enhances the ability of tissues to respond to subsequent sugar signals. PMID- 11439130 TI - Isolation and characterization of signal transduction mutants of Arabidopsis thaliana that constitutively activate the octadecanoid pathway and form necrotic microlesions. AB - Thionins are a group of antimicrobial polypeptides that form part of the plant's defense mechanism against pathogens. The Thi 2.1 thionin gene of Arabidopsis thaliana has been shown to be inducible by jasmonic acid (JA), an oxylipin-like hormone derived from oxygenated linolenic acid and synthesized via the octadecanoid pathway. The JA-dependent regulation of the Thi 2.1 gene has been exploited for setting up a genetic screen for the isolation of signal transduction mutants that constitutively express the Thi 2.1 gene. Ten cet mutants have been isolated which showed a constitutive expression of the thionin gene. Allelism tests revealed that they represent at least five different loci. Some mutants are dominant, others recessive, but all cet mutations behaved as monogenic traits when backcrossed with Thi 2.1-GUS plants. Some of the mutants overproduce JA and its bioactive precursor 12-oxophytodienoic acid (OPDA) up to 40-fold while others have the same low levels as the control wildtype plants. Two of the mutants showed a strong induction of both the salicylic acid (SA)- and the JA-dependent signaling pathways, while the majority seems to be affected only in the octadecanoid pathway. The Thi 2.1 thionin gene and the Pdf 1.2 defensin gene are activated independently, though both are regulated by JA. The cet-mutants, except for one, also show a spontaneous leaf cell necrosis, a reaction often associated with the systemic acquired resistance (SAR) pathway. PMID- 11439131 TI - Environmentally sensitive, SA-dependent defense responses in the cpr22 mutant of Arabidopsis. AB - To investigate the signaling pathways through which defense responses are activated following pathogen infection, we have isolated and characterized the cpr22 mutant. This plant carries a semidominant, conditional lethal mutation that confers constitutive expression of the pathogenesis-related (PR) genes PR-1, PR 2, PR-5 and the defensin gene PDF1.2. cpr22 plants also display spontaneous lesion formation, elevated levels of salicylic acid (SA) and heightened resistance to Peronospora parasitica Emco5. The cpr22 locus was mapped to chromosome 2, approximately 2 cM telomeric to the AthB102 marker. By analyzing the progeny of crosses between cpr22 plants and either NahG transgenic plants or npr1 mutants, all of the cpr22-associated phenotypes except PDF1.2 expression were found to be SA dependent. However, the SA signal transducer NPR1 was required only for constitutive PR-1 expression. A cross between cpr22 and ndr1-1 mutants revealed that enhanced resistance to P. parasitica is mediated by an NDR1 dependent pathway, while the other cpr22-induced defenses are not. Crosses between either coi1-1 or etr1-1 mutants further demonstrated that constitutive PDF1.2 expression is mediated by a JA- and ethylene-dependent pathway. Based on these results, the cpr22 mutation appears to induce its associated phenotypes by activating NPR1-dependent and NPR1-independent branches of the SA pathway, as well as an ethylene/JA signaling pathway. Interestingly, the SA-dependent phenotypes, but not the SA-independent phenotypes, are suppressed when cpr22 mutants are grown under high humidity. PMID- 11439132 TI - Antisense inhibition of protein phosphatase 2C accelerates cold acclimation in Arabidopsis thaliana. AB - Two related protein phosphatases 2C, ABI1 and AtPP2CA have been implicated as negative regulators of ABA signalling. In this study we characterized the role of AtPP2CA in cold acclimation. The pattern of expression of AtPP2CA and ABI1 was studied in different tissues and in response to abiotic stresses. The expression of both AtPP2CA and ABI1 was induced by low temperature, drought, high salt and ABA. The cold and drought-induced expression of these genes was ABA-dependent, but divergent in various ABA signalling mutants. In addition, the two PP2C genes exhibited differences in their tissue-specific expression as well as in temporal induction in response to low temperature. To elucidate the function of AtPP2CA in cold acclimation further, the corresponding gene was silenced by antisense inhibition. Transgenic antisense plants exhibited clearly accelerated development of freezing tolerance. Both exposure to low temperature and application of ABA resulted in enhanced freezing tolerance in antisense plants. These plants displayed increased sensitivity to ABA both during development of frost tolerance and during seed germination, but not in their drought responses. Furthermore, the expression of cold-and ABA-induced genes was enhanced in transgenic antisense plants. Our results suggest that AtPP2CA is a negative regulator of ABA responses during cold acclimation. PMID- 11439133 TI - Unexpected roles for cryptochrome 2 and phototropin revealed by high-resolution analysis of blue light-mediated hypocotyl growth inhibition. AB - Blue light (BL) rapidly and strongly inhibits hypocotyl elongation during the photomorphogenic response known as de-etiolation, the transformation of a dark grown seedling into a pigmented, photoautotrophic organism. In Arabidopsis thaliana, high-resolution studies of hypocotyl growth accomplished by computer assisted electronic image capture and analysis revealed that inhibition occurs in two genetically independent phases, the first beginning within 30 sec of illumination. The present work demonstrates that phototropin (nph1), the photoreceptor responsible for phototropism, is largely responsible for the initial, rapid inhibition. Signaling from phototropin during the curvature response is dependent upon interaction with NPH3, but the results presented here demonstrate that NPH3 is not necessary for phototropin-dependent growth inhibition. Activation of anion channels, which transiently depolarizes the plasma membrane within seconds of BL, is an early event in the cryptochrome signaling pathway leading to a phase of growth inhibition that replaces the transient phototropin-dependent phase after approximately 30 min of BL. Surprisingly, cry1 and cry2 were found to contribute equally and non-redundantly to anion-channel activation and to growth inhibition between 30 and 120 min of BL. Inspection of the inhibition kinetics displayed by nph1 and nph1cry1 mutants revealed that the cryptochrome phase of inhibition is delayed in seedlings lacking phototropin. This result indicates that BL-activation of phototropin influences cryptochrome signaling leading to growth inhibition. Mutations in the NPQ1 gene, which inhibit BL-induced stomatal opening, do not affect any aspect of the growth inhibition within the first 120 min examined here, and NPQ1 does not affect the activation of anion channels. PMID- 11439134 TI - Phosphatidic acid activates a wound-activated MAPK in Glycine max. AB - Many plant species demonstrate a systemic increase in phosphatidic acid (PA) levels after being wounded (Lee et al., 1997). To understand the role of PA in wound signal transduction, we investigated if PA can activate protein kinases in soybean (Glycine max L.). We found that a MAPK is activated in soybean seedlings in both wounded and neighboring unwounded leaves. The wound-activated soybean kinase is specifically recognized by an antibody against the alfalfa MAPK, SIMK. When PA production is inhibited with n-butanol, an inhibitor of phospholipase D, the wound-induced activation of the MAPK is suppressed, suggesting that an elevation in PA levels is essential for its activation. Supporting this is the observation that exogenous PA activates the MAPK in suspension-cultured soybean cells. Activation of the 49 kDa MAPK occurs almost exclusively by PA, as other lipids are unable to or can only weakly activate the kinase. PA-induced activation of the MAPK is not a direct effect on the kinase but is mediated by upstream kinases. Our results suggest that PA acts as a second messenger in wound induced MAPK signaling in plants. PMID- 11439135 TI - Identification of a cis-regulatory element for L1 layer-specific gene expression, which is targeted by an L1-specific homeodomain protein. AB - The Arabidopsis thaliana PROTODERMAL FACTOR1 (PDF1) gene encoding a putative extracellular proline-rich protein is exclusively expressed in the L1 layer of shoot apices and the protoderm of organ primordia. In order to identify essential cis-regulatory sequences required for the L1 layer-specific expression, a series of 5' deletions of the PDF1 promoter were fused to the beta-glucronidase (GUS) gene and introduced into Arabidopsis plants. Our analysis revealed that the minimum region necessary to confer L1-specific expression of PDF1 is confined within a 260-bp fragment upstream of the transcription start site. We identified an 8-bp motif in this region that is conserved between promoter regions of all the L1-specific genes so far cloned, and we designated it the L1 box. Electrophoretic mobility shift assays demonstrated that the L1-specific homeodomain protein ATML1 can bind to the L1 box sequence in vitro. The GUS expression in transgenic plants disappeared when a mutation that abolishes binding of ATML1 was introduced into the PDF1 l1 box sequence of the construct. These results suggest that the L1 box plays a crucial role in the regulation of PDF1 expression in L1 cells and that ATML1 could cooperate to drive L1-specific expression. PMID- 11439136 TI - Identification of a novel pathogen-responsive element in the promoter of the tobacco gene HSR203J, a molecular marker of the hypersensitive response. AB - The tobacco gene, HSR203J, which is specifically activated during the early steps of incompatible plant/pathogen interactions has been shown to be a molecular marker of the hypersensitive response (HR). It constitutes an ideal model for the identification of HR-responsive cis-regulatory elements. As a first step in the promoter dissection, deletion mutants of the 5' flanking sequence of HSR203J fused to the GUS reporter gene were analyzed. Then, the construction and study of chimeric constructs containing HSR203J promoter fragments fused to a minimal promoter enabled us to identify a 28-bp regulatory element located between -106 and -79 upstream of the transcription initiation site. This element has been shown to be necessary and sufficient for transcriptional activation in response to pathogen. It contains a 10-bp palindrome followed by its imperfect repeat. The mutagenesis of these two sequence elements led to the identification of a 12-bp motif termed HSRE (HSR203 responsive element) responsible for the marked induction of the HSR203J gene during the HR. Since this DNA region did not show any homology with known regulatory sequences, this 12 bp motif corresponds to a novel cis-regulatory element. PMID- 11439137 TI - Resistance to Pseudomonas syringae conferred by an Arabidopsis thaliana coronatine-insensitive (coi1) mutation occurs through two distinct mechanisms. AB - A new allele of the coronatine-insensitive locus (COI1) was isolated in a screen for Arabidopsis thaliana mutants with enhanced resistance to the bacterial pathogen Pseudomonas syringae. This mutant, designated coi1-20, exhibits robust resistance to several P. syringae isolates but remains susceptible to the virulent pathogens Erisyphe and cauliflower mosaic virus. Resistance to P. syringae strain PstDC3000 in coi1-20 plants is correlated with hyperactivation of PR-1 expression and accumulation of elevated levels of salicylic acid (SA) following infection, suggesting that the SA-mediated defense response pathway is sensitized in this mutant. Restriction of growth of PstDC3000 in coi1-20 leaves is partially dependent on NPR1 and fully dependent on SA, indicating that SA mediated defenses are required for restriction of PstDC3000 growth in coi1-20 plants. Surprisingly, despite high levels of PstDC3000 growth in coi1-20 plants carrying the salicylate hydroxylase (nahG) transgene, these plants do not exhibit disease symptoms. Thus resistance to P. syringae in coi1-20 plants is conferred by two different mechanisms: (i) restriction of pathogen growth via activation of the SA-dependent defense pathway; and (ii) an SA-independent inability to develop disease symptoms. These findings are consistent with the hypotheses that the P. syringae phytotoxin coronatine acts to promote virulence by inhibiting host defense responses and by promoting lesion formation. PMID- 11439138 TI - Eukaryotic features of the Xanthomonas type III effector AvrBs3: protein domains involved in transcriptional activation and the interaction with nuclear import receptors from pepper. AB - The AvrBs3 protein of the phytopathogenic bacterium Xanthomonas campestris pv. vesicatoria is targeted to host-plant cells by the bacterial Hrp type III secretion system. In pepper plants containing the Bs3 resistance gene, AvrBs3 induces the hypersensitive response (HR). AvrBs3 recognition is thought to occur in the plant cell nucleus as HR induction is dependent on nuclear localization signals (NLSs) and an acidic transcription activation domain (AAD). In a search for AvrBs3-interacting pepper proteins using the yeast two-hybrid system, we have isolated eight different classes of cDNA inserts including two genes for importin alpha proteins. Importin alpha is part of the nuclear import machinery and interacts with AvrBs3 through an NLS in the carboxy-terminus of the protein, both in yeast and in vitro. The mechanism of AvrBs3 recognition was further studied by analysis of the C-terminal AAD. This putative transcription-activation domain was shown to be required for AvrBs3 HR-inducing activity, and could be functionally replaced with the VP16 AAD from the Herpes simplex virus. Our data support the model in which the AvrBs3 effector localizes to the nucleus, where the Bs3 mediated surveillance system of resistant plants detects AvrBs3 through its interference with host gene transcription. PMID- 11439139 TI - Form I Rubiscos from non-green algae are expressed abundantly but not assembled in tobacco chloroplasts. AB - Non-green algae have Rubiscos that are phylogenetically distinct from their counterparts in green algae and higher plants. Some non-green-algal Rubiscos are more specific for CO2, relative to O2, than higher-plant Rubiscos, sometimes coupled with lower Michaelis constants for CO2. If these Rubiscos could be substituted for the higher-plant enzyme, and if they functioned successfully in the higher-plant chloroplast and were regulated appropriately, they would improve the CO2 use and quantum efficiency of higher-plant photosynthesis. To assess the feasibility of expressing non-green algal Rubiscos in higher-plant chloroplasts, we inserted the rbcLS operons from the rhodophyte Galdieria sulphuraria and the diatom Phaeodactylum tricornutum into the inverted repeats of the plastid genome of tobacco, leaving the tobacco rbcL gene unaltered. Homoplasmic transformants were selected. The transgenes directed the synthesis of abundant amounts of transcripts and both subunits of the foreign Rubiscos. In some circumstances, leaves of the transformants with the P. tricornutum Rubisco contained as much foreign Rubisco protein as endogenous tobacco Rubisco (>30% of the soluble leaf protein). However, the subunits of the foreign Rubiscos were not properly folded and/or assembled. All the foreign large subunits and most of the foreign small subunits were recovered in the insoluble fractions of leaf extracts. Edman sequencing yielded the expected N-terminal sequences for the foreign small subunits but the N-termini of the foreign large subunits were blocked. Accumulation of large amounts of denatured foreign Rubisco in the leaves, particularly of the P. tricornutum transformants, caused a reduction in the amount of tobacco Rubisco present, with concomitant reductions in leaf CO2 assimilation and plant growth. PMID- 11439140 TI - The Arabidopsis compact inflorescence genes: phase-specific growth regulation and the determination of inflorescence architecture. AB - During their life cycle, higher plants pass through a series of growth phases that are characterized by the production of morphologically distinct vegetative and reproductive organs and by different growth patterns. Three major phases have been described in Arabidopsis: juvenile vegetative, adult vegetative, and reproductive. In this report we describe a novel, phase-specific mutant in Arabidopsis, compact inflorescence (cif). The most apparent aspect of the cif phenotype is a strong reduction in the elongation of internodes in the inflorescence, resulting in the formation of a floral cluster at the apical end of all reproductive shoots. Elongation and expansion of adult vegetative rosette leaves are also compromised in mutant plants. The onset of the cif trait correlates closely with morphological changes marking the phase transition from juvenile to adult, and mutant plants produce normal flowers and are fully fertile. Hence the cif phenotype appears to be adult vegetative phase-specific. Histological sections of mutant inflorescence internodes indicate normal tissue specification, but reduced cell elongation compared to wild-type. compact inflorescence is inherited as a two-gene trait involving the action of a recessive and a dominant locus. These two cif genes appear to be key components of a growth regulatory pathway that is closely linked to phase change, and specifies critical aspects of plant growth and architecture including inflorescence internode length. PMID- 11439141 TI - Introduction to the series. Danger model of immunity. PMID- 11439142 TI - Essay 1: the Danger model in its historical context. PMID- 11439143 TI - Genetic dissection of age-related changes of immune function in mice. AB - Understanding of the genetic basis of normal and abnormal development of the immune response is an enormous undertaking. The immune response, at the most minimal level, involves interactions of antigen presenting cells (APCs), T and B cells. Each of these cells produce cell surface and soluble factors (cytokines) that affect both autocrine and paracrine functions. A second level of complexity needs to consider the development of the macrophage/monocyte lineage as well as the production of the common lymphoid precursor which undergoes distinct maturation steps in the thymus and periphery to form mature T cells as well as in BM (BM) and lymphoid organs to form mature B cells. A third level of complexity involves the immune response to infectious agents including viruses and also the response to tumour antigens. In addition, there are imbalances that predispose to decreased responses (immunodeficiencies) or increased responses (autoimmunity). A fourth level of complexity involves attempts to understand the differences in the immune response that occurs at a very young age, in adults, and at a very old age. This review will focus on the use of C57BL/6 J X DBA/2 J (BXD) recombinant inbred (RI) strains of mice to map genetic loci associated with the production of lymphoid precursors in the BM, development of T cells in the thymus, and T-cell responses to stimulation in the peripheral lymphoid organs in adult and in aged mice. Strategies to improve the power and precision in which complex traits such as the age-related immune response can be mapped is limited with the current set of 35 strains of BXD mice. Strategies to increase these strains by generating recombinant intercross (RIX) strains of mice are being developed to enable this large set of lines to detect quantitative trait loci (QTLs) with a much higher consistency and statistical power. More importantly, the resolution with which these QTLs can be mapped would be greatly improved and, in many cases, adequate to carry out direct identification of candidate genes. It is likely that, given the complexity of the immune system development, the number of cells involved in an immune response, and especially the changes in the immune system with ageing, mapping hundreds of genes will be required to fully understand age-related changes in the immune response. This review outlines ongoing and future strategies that will enable the mapping and identification of these genes. PMID- 11439144 TI - Non-Th2 regulatory T-cell control of Th1 autoimmunity. AB - The Th1/Th2 concept brought an attractive explanation of the active self tolerance which appears to control the onset of pathogenic autoimmunity. New data coming from various independent horizons indicate that self immunoregulation could also depend to a large extent on non-Th2 cells. Original data derived from the day-3-thymectomy model, selective T-cell lymphocytopenia and nonobese diabetic mice are discussed in an effort to analyze similarities and differences in phenotype (CD25, CD62L and CD45RB) and cytokine pattern (notably interleukin (IL)-4, IL-10 and transforming growth factor (TGF)beta) of regulatory cells involved in these models. The relationship of these cells with Th3, Tr1 and natural killer (NK) T cells are also discussed. The hypothesis is proposed that CD25 CD62L T cells mediate the physiologic regulation of self regulation whereas Th2 and Th3 cells are essentially induced following sensitization against autoantigens. PMID- 11439145 TI - Recent progress in the understanding of B-cell functions in autoimmunity. AB - Our early concepts of the normal role of B cells in immunity focused on their ability to produce antibodies (Ab) and in the case of autoimmune diseases autoAbs, some of which were pathogenic. Over the past 10 years, it has became apparent that B cells display a variety of characteristics, other than Ab production, which could contribute to autoimmunity. They normally play a role in the development of lymphoid architecture, regulating T-cell subsets and dendritic cell (DC) function through cytokine production, and in activation of T cells. Receptors editing is also important in B cells which aids in immunity to infection and, possibly, prevention of autoimmunity. Transgenic animal models have now shown that B cells are necessary for many autoimmune diseases although their Ab products are not required in some cases. Negative signalling by CD5 and other molecules, such as CD22, in maintaining tolerance through recruitment of src-homology two domain-containing protein tyrosine phosphatase-1 has also been documented. In fact, we have now reached a new era whereby the B cell has returned as an important contributor to autoimmune disorders, so that the race is on to characterize signalling regulation via the B-cell receptor and coreceptors. Identification of such molecules and their potential defects should lead to effective ways of controlling the immune response and in particular preventing the development of autoimmune states. The classical view of B cells in the biology of immune responses to infectious and self-antigens (Ag) that they promote immunity primarily by producing Ab turns out to be rather naive. Indeed, studies over the last few years indicate that this view is far from complete, and suggest that B lymphocytes have extraordinarily diverse functions within the immune system. Furthermore, it is becoming increasingly clear that the pathogenesis of autoimmune diseases cannot solely be accounted for by T cells, and intrinsic abnormalities of B cells have been described in such conditions. In this brief review we highlight some recent observations in the context of B lymphocyte in pathophysiology, and focus on their revival as pivotal players the pathophysiology in autoimmune diseases. Yet, it remains difficult to provide a model of how important B cells are in immunity and autoimmunity. PMID- 11439146 TI - MHC class II allosteric site drugs: new immunotherapeutics for malignant, infectious and autoimmune diseases. AB - The discovery of the interactions of the 'Ii-Key' segment of the Ii protein with the major histocmpatibility complex (MHC) Class II allosteric site, which is adjacent to the antigenic peptide-binding site, creates therapeutic opportunities by regulating the antigenic peptide binding to MHC class II molecules. The binding of Ii-Key to the MHC class II allosteric site loosens the hold of the MHC Class II 'clamshell' on antigenic peptides and leads to highly efficient antigenic peptide charging to or releasing from the MHC class II antigenic peptide-binding groove. Ii-Key peptide-induced spilling of bound antigenic peptide, or replacement with inert blockers, leads to 'inert immunosuppression'. Highly efficient replacement of ambient with vaccine peptides by Ii-Key permits 'active immunosuppression' for antigen-specific control of autoimmune diseases in the absence of cytokines or adjuvants. On the other hand, active immunization against cancer or infectious disease can result from epitope replacement mediated by Ii-Key and accompanied by cytokines or other adjuvants. Finally, linking the Ii-Key peptide through a simple polymethylene bridge to an antigenic sequence vastly increases the potency of MHC Class II peptide vaccines. In summary, the discovery of the MHC class II allosteric site allows one to increase the efficiency of MHC class II-related, antigenic epitope-specific therapy for malignant, infectious, and autoimmune diseases. The focus of this review is on the mechanism and potential clinical use of such novel allosteric site-directed, Ii-key drugs. PMID- 11439147 TI - Key sequences involved in the spreading of the systemic autoimmune response to spliceosomal proteins. AB - Immune spreading to multiple intracellular antigens is likely to be of primary importance in organ-specific and systemic autoimmune diseases. A number of mechanisms by which immune spreading may occur from only a single autoreactive epitope have been proposed. Search for an initiator or early epitope thus represents an important area of investigation. For example, many studies have focused on the identification of epitopes recognized by the antibodies from both patients with systemic lupus erythematosus (SLE) and lupus-prone mice. Recently, an autoepitope present in the 70K U1 ribonucleo protein (RNP) and recognized by CD4+ T cells from lupus mice has also been identified. Here, we analyze the results of B- and T-cell-epitope mapping studies of several RNPs present in the spliceosome and propose a model of epitope spreading. In this model, a consensus sequence (the RNP motif) conserved in many nuclear, nucleolar and cytoplasmic antigens, might play a role as 'driver' epitope. This hypothesis is based on the observation that this sequence is recognized by CD4+ T cells from lupus mice and is often targeted by autoantibodies, very early during the course of the disease. Targeting this region that is repeated in different self-antigens, might represent an interesting strategy to interfere with the continuous T-cell stimulation and exposure to specific antigens. PMID- 11439148 TI - Cytokine polymorphisms in systemic lupus erythematosus and Sjogren's syndrome. AB - Systemic lupus erythematosus (SLE) and Sjogren's syndrome (SS) are defined genetically as complex diseases where multiple genes are involved in their pathogenesis. Among the genes of interest are those coding for the cytokines, molecules involved in immunoregulation that have been shown to play important roles in these diseases. Whether abnormalities in cytokine production are owing to genetic polymorphisms within the genes themselves is a matter of intensive study. The finding of functional polymorphisms within cytokine genes and their potential association with disease will open new avenues in their treatment. PMID- 11439149 TI - Chemokines and cell trafficking in Sjogren's syndrome. AB - Sjogren's syndrome is a chronic inflammatory condition affecting exocrine glands, manifested clinically as dry eyes and dry mouth. It arises secondary to systemic immune-mediated diseases such as rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), scleroderma or 'primary' Sjogren's syndrome. Histologically it is characterized by peri-ductal aggregates of CD4 T lymphocytes, the frequent occurrence of ectopic germinal centres and, in some patients, B-cell infiltration of ductal epithelium (myoepithelial sialadenitis). This latter lesion is the precursor for the development of low grade (MALT) B-cell lymphoma. The identification over recent years of chemokines and their receptors enables us to address the specific processes involved in the migration of inflammatory cells into exocrine glands, the development of their secondary structures and patterns of retention within the glands and potentially the subsequent transformation of B cells into mucosa associated lymphoid tissue (MALT) lymphoma. PMID- 11439150 TI - Adhesive interactions between human NK cells and porcine endothelial cells. AB - Human natural killer (NK) cells are able to adhere to xenogeneic porcine endothelial cells (EC) and evidence from in vitro studies as well as animal models suggests a potential role for NK cells in the cellular recognition and damage of porcine xenogeneic tissues. One possible explanation for the observed NK cell-mediated xenogeneic cytotoxicity against porcine EC is the molecular incompatibility between porcine major histocompatibility complex (MHC) class I molecules and MHC-specific inhibitory receptors on human NK cells. In this review we attempt to summarize the current knowledge concerning adhesive interactions between human NK cells and porcine EC under special considerations of the cross species receptor-ligand interactions. Methodological differences in assessing adhesion between various studies are reviewed and comparisons to the syngeneic/allogeneic adhesion mechanisms are made. Finally, the therapeutic potential of blocking antibodies and transgenic HLA expression in preventing NK cell adhesion and xenogeneic cytotoxicity is discussed. PMID- 11439151 TI - Heat shock protein 60 specific T-cell response in chlamydial infections. AB - Heat shock proteins (HSPs) of most pathogens, including Chlamydia, are major immune targets of both humoral- and cell-mediated immune mechanisms. During the last decade, many investigators have focused their research to elucidate the complex relationship of chlamydial HSPs, especially chlamydial HSP60, and the host immune response. A central issue is whether the pathologic mechanisms in chronic chlamydial diseases are associated with an enhanced immune response to chlamydial HSP60 which can mediate tissue destruction through cytotoxic reactions, or whether they are related to the Th2 type of response that eventually leads to partial or temporary suppression of an effective antichlamydial response. Our review highlights the available knowledge between immune responses to chlamydial HSP60 and chronic chlamydial infections in human. PMID- 11439152 TI - Liberation of soluble proteins from live and dead mycobacterial cells and the implications for pathogenicity of tubercle bacilli hypothesis. AB - Soluble proteins liberated from live M. tuberculosis are translocated through the cytoplasmic membrane to a 'periplasmic space'. For further export of proteins across the outer permeability barrier, it is necessary to postulate an excretion mechanism possibly involving some kind of porin. Observations of the repertoire of proteins in culture filtrates after liquid culture of M. tuberculosis show that a large repertoire of various kinds of proteins cross the outer permeability barrier of tubercle bacilli indicating that the excretion mechanism has a wide range of specificities for proteins. Culture filtrates of tubercle bacilli almost always contain both truly secreted proteins and cytoplasmatically-derived proteins. It is questionable whether cytoplasmic proteins can cross an intact cytoplasmic membrane. The simplest explanation for the appearance of cytoplasmic proteins in culture filtrates of tubercle bacilli would be that they are released after disintegration of the cytoplasmic membrane in dying or dead bacilli. Tubercle bacilli armed with secreted factors that may specifically inhibit innate and adaptive immune responses, excrete these from the periplasmic space of live bacilli. Unspecific in its character, the excretion mechanism also liberates proteins that are essential for building and maintaining the cell wall, thereby reducing the effectiveness of this process. This may be part of the explanation why M. tuberculosis and other pathogenic mycobacteria grow so slowly. Finally, it may be postulated that dormant or latent tubercle bacilli use their repertoire of secreted proteins to control their intracellular habitat and that bacterial cytoplasmic proteins would not be liberated from such bacilli. The consequence would be that only immune responses to secreted proteins would be effective for elimination of the dormant stage of infection. In a situation with active infection there will be considerable growth and turnover of bacilli with liberation of all kinds of immunogenic substances from the bacilli. In this situation immunity against cytoplasmic proteins would also be effective and immunity to cytoplasmic proteins should also be effective for control of the reactivation of latent disease because as soon as the bacilli start to grow there will also be a subpopulation of dead bacilli on the arena. PMID- 11439153 TI - Signalling via MHC class II molecules modifies the composition of GEMs in APC. AB - Major histocompatibility complex (MHC) class II molecules are responsible for peptide presentation to helper T lymphocytes and as such play an essential role in the immune response. These molecules transmit intracellular signals leading to diverse consequences in B lymphocytes including proliferation and apoptosis. Recent studies have revealed that glycolipid enriched membrane microdomains (GEMs) behave as signalling platforms for a variety of lymphocyte receptors. We have quantified human leucocyte antigen (HLA)-DR molecules localized in GEMs in human B lymphocytes. Use of a model imitating the interaction of HLA-DR with a T cell receptor (TCR) modified the constituents of the HLA-DR-enriched GEMs. Confocal microscopy demonstrated a recruitment of HLA-DR and the ganglioside GM1 at the site of HLA-DR interaction with the stimulating ligand. Moreover, cholesterol depletion efficiently impaired this recruitment. Co-localizing proteins detected in HLA-DR-enriched GEMs include protein kinase C (PKC)-delta and actin. These data reveal that MHC class II antigens are localized in GEMs in mature human B lymphocytes and indicates that the formation of the immunological synapse regulates the composition of HLA-DR enriched GEMs in the antigen presenting cell (APC). PMID- 11439154 TI - Decreased T cell stimulatory capacity of monocyte-derived human macrophages following herpes simplex virus type 1 infection. AB - Macrophages play a central role in establishing a specific immune response by acting as professional antigen presenting cells (APC) for T cells leading to a vigorous immune response. In order to analyze if Herpes simplex Virus (HSV) type 1 infection might affect the macrophage APC-function, monocyte-derived human macrophages were infected with HSV-1 strain F in vitro. Cocultures with allogeneic T cells revealed a strongly impaired stimulatory capacity of HSV infected macrophages compared to uninfected controls which was not owing to a productive viral infection in macrophages. An increased expression of Fas ligand (FasL/CD95L) was detected in HSV-infected macrophages by FACS analysis. Although the majority of the macrophages expressed high levels of Fas (CD95/Apo-1), the HSV-induced upregulation of FasL did not result in an increased autocrine apoptosis of macrophages which might be related to endogenous expression of the apoptosis inhibitor FLICE inhibitory protein (FLIP). However, substantial apoptosis occurred in peripheral T cells as well as Fas-sensitive Jurkat T cells when cocultured with HSV-infected macrophages. These findings suggest that the paracrine killing of activated T cells by FasL expressing APC might be a novel strategy of immune evasion by HSV. PMID- 11439155 TI - Switch in chemokine receptor phenotype on memory T cells without a change in the cytokine phenotype. AB - Th1 and Th2 cells as defined by their cytokine profile are associated with the expression of the chemokine receptors CCR5 and CCR3, respectively. In committed human memory Th1 cells the cytokine profile is irreversibly expressed. However, it is not known if the chemokine receptor phenotypes of Th1 and Th2 cells are permanently associated to the cytokine profile or if it can be changed. To analyze the possibility of inducing a switch in chemokine receptor phenotype on memory Th cells we used differentiated memory Th cells isolated from synovial tissue (ST) samples of patients with rheumatoid arthritis (RA). Freshly isolated T cells, T-cell lines and T-cell clones from these tissues were manipulated with Th1 (interleukin (IL)-12 + anti IL-4) or Th2 (IL-4 + anti IL-12) inducing conditions. The surface expression of CCR5 and CCR3 was analyzed by flowcytometry and interferon (IFN)-gamma and IL-4 production by ELISA. A Th1-inducing cytokine environment increased the expression of CCR5 in Th1 cells and induced the expression of CCR5 in Th2 cells as compared to culture condition with only IL-2. Induction of CCR5 expression on Th2 clones was associated with secretion of some IFN-gamma. Moreover, the Th2-associated chemokine receptor CCR3 could be expressed on both Th1-dominant cell lines, and clones of Th1 and Th0 type after culture conditions with IL-4. This expression of CCR3 was associated with a reduced IFN-gamma production, but no IL-4 production could be induced. The IL-4 treated Th1 clones had a reduced migratory capacity against chemokines produced by ST cells compared to nonmanipulated T-cell clones. In contrast, the same IL-12 treated Th1 clones showed an increased migratory potential. Induction of the Th2 associated marker CCR3 on memory Th1 cells demonstrates that a change in chemokine receptor phenotype related to the Th2 type can be induced on terminally differentiated Th1 cells, without a change in the cytokine profile. PMID- 11439156 TI - Inhibitory effects of MoAbs against a surface protein antigen in real-time adherence in vitro and recolonization in vivo of Streptococcus mutans. AB - A surface protein antigen (PAc) of Streptococcus mutans, particularly the A region of the molecule, has been reported to interact with salivary components on the tooth surface. It might be a candidate antigen inducing the production of antibodies against the adherence of S. mutans to the tooth surface. We investigated the effects of monoclonal antibodies (MoAbs) obtained by immunization of synthetic PAc peptides that completely correspond to the amino acid sequence of part of the A-region. These MoAbs recognize several core B-cell epitopes in the sequence. Two (KH5 and SH2) of these antibodies reacted with both S. mutans and Streptococcus sobrinus, but not with Streptococcus sanguis, Streptococcus salivarius, Porphyromonas gingivalis or Lactobacillus casei. They clearly inhibited the real-time adherence of S. mutans to salivary components in a biosensor. KH5, which showed a real-time inhibition (71%), also significantly prevented the recolonization of S. mutans on the tooth surface in rats. These results suggested that the core B-cell epitope (-Y---L--Y----) recognized by KH5 was the essential sequence in the antigenic epitopes of PAc protein recognized specifically by the inhibitory antibody. Therefore, the amino acid residues were found to be important in the initial attachment of S. mutans to the tooth surface. These results provide for the mechanism of PAc molecule in the initial attachment of S. mutans on the tooth surface and more effective designs for the removal of S. mutans and S. sobrinus from the oral cavity. PMID- 11439157 TI - Comparative immunization study using RNA and DNA constructs encoding a part of the Plasmodium falciparum antigen Pf332. AB - Development of nucleic acid-based vaccines against parasitic diseases shows great promise, although certain concerns about safety aspects of conventional DNA vaccines have been raised. This study presents a comparison of antibody responses induced in mice by DNA and RNA-based immunization with vectors encoding a part of the P. falciparum antigen Pf332. Two types of plasmids were used, one conventional DNA plasmid containing a cytomegalovirus promoter and one suicidal DNA plasmid encoding the Semliki Forest virus (SFV) replicase. RNA, encoding the SFV replicase and the relevant antigen, was delivered either as naked RNA or packaged in SFV suicide particles. In general, the antibody responses induced by the DNA plasmids were low and peaking after three injections, the conventional plasmid giving the highest responses. Also the RNA delivered in SFV particles consistently induced antibody responses, although comparatively low. Analyses of the ratio of immunoglobulin (Ig)G1/IgG2a subclasses in the responses indicated that all plasmids resulted in a bias for a Th2-type of response, while the SFV particles elicited a Th1 type of response. Importantly, all these immunogens induced an immunological memory, which could be efficiently activated by a booster injection with the corresponding protein, with unchanged patterns of IgG subclasses. PMID- 11439158 TI - Use of liposomes as an immunopotentiating delivery system: in perspective of vaccine development. AB - Liposomes have been widely used to deliver antigens to the antigen-presenting cells (APCs) and also to modify their immunological behaviour in model animals. We recently demonstrated the potential of yeast lipid liposomes to undergo membrane-membrane fusion with cytoplasmic membrane of the target cells. Interestingly, studies in the present report revealed that antigen encapsulated in yeast lipid liposomes could be successfully delivered simultaneously into the cytosolic as well as endosomal processing pathways of APCs, leading to the generation of both CD4+ T helper and CD8+ cytotoxic T cells. In contrast, encapsulation of same antigen in egg phosphatidyl-choline (PC) liposomes, just like its free form, has inefficient access to the cytosolic pathway of major histocompatibility complex (MHC) I dependent antigen presentation and failed to generate antigen specific CD8+ cytotoxic T-cell response. However, both egg PC as well as yeast lipid liposomes have elicited strong antigen specific antibody responses in immunized animals. These results imply usage of liposome encapsulated antigen as potential candidate vaccine capable of eliciting both cell mediated as well as humoral immune responses. PMID- 11439159 TI - IL-4-dependent effector phase in autoimmune exocrinopathy as defined by the NOD.IL-4-gene knockout mouse model of Sjogren's syndrome. AB - NOD mice manifest many features of autoimmune exocrinopathy (Sjogren's syndrome), a disease generally characterized by a chronic, progressive immunological attack against the exocrine tissues of the salivary and lacrimal glands. Previous studies using the NOD congenic partner strain, NOD.Igmu(null), defined an important role for B lymphocytes in the development of xerostomia, implicating autoantibodies reactive with the acetylcholine muscarinic receptor (M3R) as the possible effector mechanism. In the present study, we have examined the impact of the cytokine, interleukin (IL)-4, on autoimmune exocrinopathy by using the IL-4 gene knockout (KO) NOD mouse strain, NOD.IL-4-/-. Despite manifesting the physiological aberrations and marked leukocytic infiltration of the salivary glands characteristic of autoimmune xerostomia in NOD mice, the NOD.IL-4-/- mice do not develop xerostomia. However, NOD.IL-4-/- mice that received adoptively transferred T lymphocytes derived from NOD.Igmu-/- mice progress to xerostomia, thereby reversing the defect. While progression or lack of progression to xerostomia correlated with the ability of the NOD.IL-4-/- mice to express detectable anti-M3R autoantibodies, the precise mechanism of how IL-4 influences the development of autoimmune xerostomia remains speculative. PMID- 11439160 TI - Structure and chromosomal localization of chicken CD5. AB - CD5 is a transmembrane glycoprotein on all T cells and on a subpopulation of B cells. Based on the analysis of chicken CD5-cDNA we have previously shown that the structure of the CD5 protein is conserved between species. Here we report the isolation and chromosomal mapping of the chicken CD5 gene. The gene spans 3.4 kb and is extremely compact with a high GC-nucleotide content. There are 10 exons and the introns are spliced out similarly to those in the human CD5 gene. Each of the three extracellular scavenger receptor cysteine-rich (SRCR) domains is encoded as an exon of its own, as is the proline-rich hinge region that separates the first two membrane-distal SRCR domains. The fluorescence in situ hybridization (FISH) technique was used to map the gene to chromosome five. This is the first report describing the organization of the CD5 gene from a nonmammalian species. PMID- 11439161 TI - Developmentally regulated expression of the transmembrane adaptor protein trim in fetal and adult T cells. AB - TRIM is a recently identified transmembrane adaptor protein which is exclusively expressed in T cells and natural killer (NK) cells. In peripheral blood T cells TRIM has been reported to coprecipitate, comodulate, and cocap with the T-cell receptor (TCR), suggesting that it is an integral component of the TCR/CD3/zeta complex. Here we investigate the expression of TRIM mRNAs and proteins in developing thymocytes. Two splicing isoforms with open reading frames are observed, namely a full length (TRIM) and a truncated version (DeltaTM-TRIM). The latter lacks the extracellular and transmembrane domains as well as the first 10 cytoplasmic aminoacids and is significantly expressed only as mRNA in early fetal thymocytes. TRIM mRNA is detected in all mainstream thymocyte subsets in adult mice. TRIM protein, in contrast, first appears in the DN2 (CD44+ CD25+) subset of adult double negative (DN) cells. In fetal thymocyte development, TRIM mRNA is seen from dg 14.5 onwards whereas TRIM protein appears first on dg 16.5. In contrast to the adult, the TRIM protein was seen in a subset of fetal DN1 cells. In fetal and adult thymocytes, TRIM protein expression was highest in DN2, DN3 (CD44-25+) and in DP cells, compatible with a functional role at or around phases of thymic selection. PMID- 11439162 TI - On the role of CD3delta chains in TCRgammadelta/CD3 complexes during assembly and membrane expression. AB - The present study was performed in order to analyze whether T-cell receptor (TCR)/CD3 assembly, intracellular transport and surface expression are carried in a similar way in alphabeta-and gammadelta-T cells. By means of optimal immunoprecipitation conditions with 35S-methionine/cysteine- or biotin-labelled TCR/CD3 proteins from alphabeta- or gammadelta-T-lymphoma-cell lines, as well as TCRgammadelta cDNA transfectants, it was found that CD3delta chains associate less strongly with TCRgammadelta heterodimers compared to TCRalphabeta heterodimers. This preferential reactivity of CD3delta chains appears to be structural and not owing to differences in gammadelta- versus alphabeta-T-cell intracellular environments. Our results are in accordance firstly, with data from CD3delta-deficient mice, which have gammadelta-T cells but no alphabeta-T cells, secondly with the suggested role of CD3delta chains in the positive selection of alphabeta-T cells, a process apparently not followed by gammadelta-T cells, and lastly with the differential roles of CD3delta chains versus CD3gamma chains, explaining the maintenance of two CD3delta and CD3gamma genes after the duplication from a CD3delta/gamma gene present in avians. The impaired reactivity of CD3delta chains with TCRgammadelta heterodimers seems to be owing to a less efficient association with TCRgamma chains. In contrast, CD3delta chains interact as strongly with TCRdelta chains as do CD3gamma chains with both TCRgamma and TCRdelta chains. These data may explain, at the molecular levels, why surface TCR/CD3 expression levels are impaired in gammadelta-T cells from CD3gamma deficient mice but not from CD3delta-deficient mice. PMID- 11439163 TI - Endogenous inhibition of antimycobacterial immunity by IL-10 varies between mycobacterial species. AB - Interleukin (IL)-10 is an immunoregulatory cytokine that inhibits both Th1-like T cell responses and macrophage activation. Deficiency of IL-10 has been associated with increased Th1-like CD4+ T-cell responses and increased clearance of some intracellular pathogens, however, its role in mycobacterial infections is controversial. In order to examine the effects of mycobacterial virulence on the outcome of infection we compared infection with Mycobacterium avium and virulent Mycobacterium tuberculosis in C57Bl/6 IL-10-/- mice. M. avium infection in IL-10 /- mice resulted in sustained increases in interferon (IFN)-gamma-secreting T cell responses and was associated with the increased clearance of M. avium from the liver and lung. By contrast, M. tuberculosis infection in IL-10-/- mice led to a transient increase in IFN-gamma T-cell responses at 4 weeks postinfection, with reduced bacterial burden in the lungs. This was not sustained so that by 8 weeks there was no difference to wild-type (WT) mice. In vitro infection of IL-10 /- macrophages with M. avium, but not M. tuberculosis, led to an increased IL-12 production. Therefore, endogenous IL-10 exerts a significant inhibition on specific IFN-gamma T-cell responses to M. avium infection, however, this effect is short lived during the M. tuberculosis infection, and fails to influence the long-term course of infection. PMID- 11439164 TI - Experimental chronic arthritis and granulomatous inflammation induced by bifidobacterium cell walls. AB - Effects of cell walls (CWs) from two almost identical strains of Bifidobacterium adolescentis were studied in rats, using three different doses. A single i.p. injection of both CWs triggered a long-lasting arthritis with CW degradation products present in the joint tissue. Histologically, the arthritis was characterized by inflammatory cells, synovial hyperplasia, pannus formation and bone erosion, closely resembling human rheumatoid arthritis (RA). In addition, CWs of the other strain induced a remarkable granuloma formation in the spleen and liver. Both CWs have the same peptidoglycan (PG) type A4alpha/beta, but differ from each other in three aspects. CW of the granuloma inducing strain: firstly has more lysine and less ornithine in PG stem peptides; secondly is more resistant to lysozyme degradation, and thirdly is better retained in the spleen. All these in comparison to the other strain used. Such characteristics are associated with the capacity to induce chronic arthritis, but it remains open how crucial they are for the granuloma formation. PMID- 11439165 TI - Characterization of the human FLICE-inhibitory protein locus and comparison of the anti-apoptotic activity of four different flip isoforms. AB - Death receptor-mediated apoptosis is involved in the regulation of immune responses and in the maintenance of immunological tolerance. FLICE-inhibitory proteins (FLIPs) are important modulators of death receptor-mediated apoptosis. To date, the FLIP family encompasses multiple members, of which some are reported to be antiapoptotic and others pro-apoptotic. This led us to investigate the activity of several FLIP proteins in vitro. Concomitant with the cloning of various FLIP isoforms, a new and unexpected member of the FLIP family, denoted FLIPR, was isolated from the human Burkitt lymphoma B-cell line Raji. During the characterization of FLIPR, the genomic sequence of human FLIP was found in the NCBI GenBank. This enabled us to present the complete exon-intron constellation of the human FLIP gene and the generation of all known human FLIP isoforms by alternative splicing. We show that the human FLIP gene with a size of approximately 48 kb, consists of at least 14 exons and can give rise to 11 distinct isoforms by alternative splicing. When studying the activity of some of these isoforms, including FLIPR, they all efficiently inhibited Fas-mediated apoptosis in A20 B lymphoma cells by impeding caspase-8, -3 and -7 activity as well as poly(ADP-ribose) polymerase (PARP) cleavage. PMID- 11439166 TI - Genetic control of resistance to mercury-induced immune/autoimmune activation. AB - Previous studies have shown that genetic factors control the susceptibility to mercury-induced immunoglobulin (Ig)G1 antibody formation, IgE synthesis, renal IgG deposits and antinucleolar autoantibodies (ANolA) production in the susceptible mice. In this study, we examined the genetic control of resistance to these characteristics after HgCl2 injection in F1 hybrid crosses between the highly mercury resistant DBA/2 and mercury susceptible NZB (H-2d), SJL (H-2 s), A.CA (H-2f) and DBA/1 (H-2q) mice and also in backcross hybrids between (DBA/2 x SJL)F1 and SJL mice. We observed that mercury-induced immune/autoimmune manifestations were profoundly downregulated in most (if not all) of the F1 hybrids, indicating that the resistance to mercury was a dominant trait. Analysis of mercury-induced immune/autoimmune responses in the (DBA/2 x SJL) x SJL backcross hybrids suggested that only one gene or a cluster of genes determined the resistance to the ANolA production, whereas the resistance to other characteristics was controlled by two and/or three gene loci. By H-2 genotyping the backcross mice, it was found that H-2d haplotype per se could confer resistance to ANolA production. However, we did not find any significant association between the H-2d haplotype and the resistance to increase of IgG1 and IgE synthesis and the development of renal IgG1 deposits. Thus, while in DBA/2 mice, gene(s) in the H-2 loci strictly contribute to the inheritance of resistance to ANolA production; non-H-2 genes mainly govern the inheritance of unresponsiveness regarding other characteristics. PMID- 11439167 TI - Long-term donor chimerism after MHC (RT1) mismatched bone marrow transplantation in the rat: the role of host alloreactive NK cells. AB - We have investigated the role of major histocompatibility complex (MHC) (RT1) disparities in the engraftment of bone marrow (BM) cells after whole body irradiation of rats. Mononuclear BM cells from PVG.RT7.2 (RT1c) rats were injected i.v. into sublethally (10Gy) whole body irradiated PVG (RT1c) rats and RT1 congenic and recombinant PVG rats. Repopulation of the BM, spleen, and blood with donor cells was assessed by FACS analysis of cells labelled with the fluorescein isothiocyanate (FITC)-labelled HIS41 monoclonal antibody (MoAb) against the RT7.2 marker. In RT1 matched (PVG.RT7.2 --> PVG) and RT1-mismatched combinations (PVG.RT7.2 --> PVG.1AV1), where radioresistant host natural killer (NK) cells could not recognize the BM inoculum as foreign, a donor chimerism close to 100% was observed after 6-8 weeks. However, in rat strain combinations where host NK cells could recognize an RT1 mismatch, almost no donor cells survived, and the rats were repopulated with leukocytes of host origin. In intra MHC recombinant rat strains the element determining rejection or acceptance of the allograft mapped to the RT1-B/D-C/E/M region in PVG.R8 and PVG.R23 rats, in accordance with the patterns of NK alloreactivity in these strain combinations. NK cells may therefore be a primary obstacle to successful allogeneic BM engraftment in this model. PMID- 11439168 TI - Structural analysis of TCRalpha and beta chains from human T-Cell clones specific for small nuclear ribonucleoprotein polypeptides Sm-D, Sm-B and U1-70 kDa: TCR complementarity determining region 3 usage appears highly conserved. AB - Systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD) are systemic autoimmune diseases that are characterized by the presence of autoantibodies reactive with U small nuclear RNP (snRNP) autoantigens. Both B and T cells are important in the pathogenesis of the disease, and T- and B-cell immunity against snRNP polypeptides have been shown to be linked in vivo. Currently, several alternative hypotheses for the pathogenesis of these diseases have been proposed. These include loss of tolerance, modified self-antigens, molecular mimicry and nondirected immune activation. To help distinguish between the various models of disease pathogenesis, we have characterized the T-cell receptor (TCR) CDR3 from a large panel of well-characterized human T-cell clones and lines specific for individual snRNP polypeptides. The results presented here reveal highly restricted TCR usage across patients by the snRNP-reactive T cells based on the deduced amino acid sequence of the CDR3 loop. These data support the hypothesis that T-cell responses against self antigens in SLE and MCTD are antigen driven and that there are a limited number of T-cell epitopes present on the snRNP autoantigens. PMID- 11439169 TI - Development of anti-dsDNA autoantibodies prior to clinical diagnosis of systemic lupus erythematosus. AB - Anti-double stranded (dsDNA) antibodies are of considerable diagnostic value and are thought to be involved in the pathogenesis of systemic lupus erythematosus (SLE). Fluctuations in anti-dsDNA antibody levels are also used as markers for disease activity and exacerbations. In this study we sought to evaluate the anti dsDNA antibody level in serum samples collected before the onset of SLE diagnosis. A total of 130 SLE patients were identified with stored serum samples available prior to diagnosis within the US Department of Defense serum repository. All 633 sera available from these patients were screened for anti dsDNA antibodies using an enzyme linked immunosorbant assay (ELISA). Within this cohort 55% of cases had detectable anti-dsDNA antibodies prior to SLE diagnosis. The onset of anti-dsDNA antibodies ranged from 9.3 years before to within the same month as diagnosis (with a mean onset 2.7 years before diagnosis). In order to assess for fluctuations in anti-dsDNA levels relative to diagnosis, cases were selected with at least two positive samples, one within 6 months and a second greater than 6 months prior to diagnosis (n = 26). Seven of these cases also had samples available shortly after diagnosis (< or = 6 months) for comparison. Fifty eight percent of the 26 cases developed a significant rise in anti-dsDNA antibody levels within 6 months of diagnosis. A significant decline in anti-dsDNA levels ensued after diagnosis (and following treatment with corticosteroids) in all seven cases with samples available. Patients with a significant rise in anti dsDNA antibodies at diagnosis were more likely to have renal disease than those who did not (66.7% compared to 27.3%, chi2 =3.94, P<0.05). These data suggest that anti-dsDNA antibodies are present in SLE patient sera much earlier than previously suspected. In addition, the data are consistent with increases in anti dsDNA levels contributing to the onset of clinical illness in some patients with SLE. PMID- 11439170 TI - Disturbances in the peripheral T-cell repertoire of patients with motor neuron disease: high levels of activation and indirect evidence of superantigen. AB - Our data on peripheral blood T cells from Motor neuron disease (MND) patients indicate major immunological disturbances linked to this disease. Both CD4+ and CD8+ T-cell subsets display an increased fraction of cells showing activation markers compared to controls, indicating an unusually high level of activity in both populations. Likewise, an increased number of T-cell expansions were noted in MND patients compared to controls, most dramatically observed in the CD4+ T cell subset, where 5/144 T-cell V genes analyzed in eight subjects turned out to be expanded in the peripheral blood. In the CD8+ T-cell subset, four out of eight MND patients had peripheral BV gene expansions, 9/144 V genes analyzed. However, the most interesting result was the observation that in three out eight MND patients, expansions concerning the same BV gene were present in both CD4+ and CD8+ subsets (BV8S1 in two and BV12S1 in one patient). Parallel expansions of BV gene restricted populations in both CD4+ and CD8+ subsets in the same individual, in an major histocompatibility complex (MHC)-unrestricted manner, are normally limited to situations where superantigens are involved. No known superantigen has to date been described with the capacity to simultaneously stimulate both BV8S1 and BV12S1, suggesting that the postulated 'MND-associated' superantigen is a hitherto undefined molecule. PMID- 11439171 TI - Immunity in the female lower genital tract and the impact of HIV infection. AB - This study investigates the distribution of immunocompetent cells in the ectocervix, and cytokine and immunoglobulin (Ig) levels in cervicovaginal secretions to determine whether they are altered in asymptomatic human immunodeficiency virus (HIV) infection. Ectocervical biopsies from 10 HIV+ and 10 presumed HIV-ve women were studied by immunocytochemistry. Levels of Igs in cervicovaginal secretions were quantified by radial immunodiffusion (RID) and cytokine levels by ELISA. HIV+ women had significantly increased numbers of CD8+ lymphocytes resulting in reversal of the CD4:CD8 ratio. There was a significant increase in the proportion of activated CD8+ HLA-DR+ and CD4+ HLA-DR + lymphocytes, but not in CD8+ TIA-1+ cells. The epithelium of the cervix from HIV+ subjects showed a significant increase in both numbers of macrophages (CD68+) and proportions of activated macrophages (CD68+ HLA-DR+) compared to normal. The stroma contained increased proportions of inductive (D1+) and suppressive (D1+ D7+) macrophages but a decrease in effector phagocyte (D7+) proportions and Langerhans' cells. Significantly lower tumour necrosis factor (TNF)-alpha levels were observed in cervicovaginal secretions from HIV+ subjects. IgG levels were 4 times higher and IgM levels twice higher in cervicovaginal secretions from HIV+ women, compared to results from normal subjects. These results suggest a response within the CD8+ cells in HIV+ women, yet these cells may have a low cytolytic capacity. The raised proportions of HLA-DR+ and D1+ CD4+ macrophages could act as antigen-presenting cells (APC) for CD4+ CD45RO+ lymphocytes, and represent a local acquired response. However, the close juxtaposition of these cells offers the potential for them to act as a local reservoir of virus and promote its proliferation. The increase of IgG over sIgA in secretions of HIV+ subjects provides evidence suggesting a dysregulation of local humoral immunity. PMID- 11439172 TI - Acute phase haemolysis in chronic cold agglutinin disease. AB - We previously described a paradoxical form of chronic cold agglutinin disease (CAD) in which haemolysis occurred during episodes of fever but only marginally during exposure to colds. In order to investigate the molecular basis for this response we performed a 12-month prospective study of a patient with CAD and paradoxical haemolysis. Blood samples were collected monthly during health, and daily following hospitalization owing to hip fracture. During health we observed decreased levels of C3, undetectable C4, a non-functional classical pathway and a normal alternative pathway. Increased concentrations of C1-INH/C1rs complexes indicated continuous formation of C1-antibody-antigen complexes. There was a low grade temperature-dependent fluctuating haemolysis as evidenced from measurements of lactate dehydrogenase. Following the hip fracture, the haemolysis increased. Levels of interleukin (IL)-1beta, IL-6, interferon (IFN)-gamma and tumour necrosis factor (TNF)-alpha increased as did C1-INH, C3, C4, CRP, and lactate dehydrogenase. The results support our hypothesis stating that paradoxical haemolysis in CAD is controlled by the availability of early classical pathway complement molecules and that haemolysis following acute phase responses occurs as a consequence of increased complement synthesis. PMID- 11439173 TI - High prevalence of influenza specific antibody secreting cells in nasal mucosa. AB - Secretory immunoglobulin A (SIgA) provides the first line of defence against pathogens initiating infection via the mucosal route, e.g. the influenza virus. The aim of this study was to examine the basal level of influenza-specific antibody-secreting cell (ASC) in the local mucosa of the upper respiratory tract. Nineteen patients scheduled for tonsillectomy were enrolled for the study, and they had not experienced influenza during the previous year. Tonsils, blood, oral fluid and a nasal biopsy were sampled, and the basal levels of ASC and antibodies (Abs) were determined. We found low numbers of influenza-specific ASC in the blood and tonsils, but there were about 10-100 times higher numbers of specific ASC in the nasal mucosa tissue despite no recent influenza exposure. Thus, the basal level of influenza-specific ASC in the mucosa of the respiratory tract may be important in the protection against influenza infection. PMID- 11439174 TI - Association of mesenchymal cells and immunoglobulins with differentiating epithelial cells. AB - BACKGROUND: Mesenchymal-epithelial interactions play an important role in the physiology and pathology of epithelial tissues. Mesenchymal cells either associate with epithelium basement membrane [pericytes and perivascular monocyte derived cells (MDC)] or reside within epithelium (MDC and T cells). Although intraepithelial mesenchymal cells were suggested to contribute to the epithelium physiology, their association with particular steps in differentiation of epithelial cells, interactions among themselves, and their fate remain unclear. We studied epitopes of mesenchymal cells and their products (immunoglobulins) in stratified epithelium of uterine ectocervix, which is one of the prototypes of complete cellular differentiation from stem into the aged cells. RESULTS: Perivascular CD14 primitive MDC associated with basal (stem) epithelial cells. Thy-1 pericytes of microvasculature secreted intercellular vesicles, which associated with Ki67 postmitotic epithelial cells expressing MHC class I. Intraepithelial T cells showed an association with veiled type MDC [dendritic cell (DC) precursors] among parabasal cells, and exhibited fragmentation after entering intermediate (mature) epithelial layers. Mature DC secreted CD68 and exhibited fragmentation after reaching mid intermediate layers. Binding of IgM was detected at the top of each layer: in the upper parabasal, upper intermediate, and most surface epithelial cells. IgG was confined to the entire superficial layer. CONCLUSIONS: These data suggest that the phylogenetically and ontogenetically developed hierarchy of mesenchymal cells (MDC, pericytes, T cells) and immunoglobulins (IgM, IgG) accompanies differentiation of epithelial cells from immature into the mature and aged phenotype. Further studies of an involvement of mesenchymal cells in the regulation of tissue homeostasis may bring novel approaches to the prevention and therapy of tissue dysfunctions characterized by permanent tissue immaturity (muscular dystrophy) or accelerated aging (degenerative diseases). PMID- 11439175 TI - The CRASH trial protocol (Corticosteroid randomisation after significant head injury) [ISRCTN74459797]. AB - BACKGROUND: Worldwide, millions of people are treated each year for significant head injury. A substantial proportion die, and many more are disabled. If short term corticosteroid infusion could be reliably shown to reduce these risks by just a few percent then this might affect the treatment of a few hundred thousand patients a year, protecting thousands from death or long term disability. STUDY DESIGN: CRASH is a large simple, placebo-controlled trial of the effects of a 48 hour infusion of corticosteroids on death and on neurological disability, among adults with head injury and some impairment of consciousness. Head injured patients with impaired consciousness who are judged to be 16 years or older are eligible if the responsible doctor is, for any reason, substantially uncertain whether or not to use corticosteroids. ORGANISATION: The CRASH trial will determine reliably the effects on death and disability of a short corticosteroid infusion following significant head injury. To detect or refute improvements of only a few percent in outcome, many thousands of acute head injury patients must be randomised between control and steroid infusions. Such large numbers will be possible only if hundreds of doctors and nurses can collaborate in the participating emergency departments. Since they are busy, and working in emergency situations, the trial involves them in almost no extra work: no special investigations or changes to usual management are required, and data collection is absolutely minimal. The trial is on-going and new collaborators are welcome. Further information about the trial is available at http://www.crash.lshtm.ac.uk PMID- 11439176 TI - Phoxy lipids: revealing PX domains as phosphoinositide binding modules. PMID- 11439177 TI - Regulation of GSK-3: a cellular multiprocessor. PMID- 11439178 TI - AMPA receptor trafficking and the control of synaptic transmission. PMID- 11439179 TI - The evolving concept of a stem cell: entity or function? PMID- 11439180 TI - The genetics and biology of vertebrate sex determination. PMID- 11439181 TI - Alan Wolffe (1959-2001). PMID- 11439182 TI - Activation of cardiac gene expression by myocardin, a transcriptional cofactor for serum response factor. AB - Serum response factor (SRF) regulates transcription of numerous muscle and growth factor-inducible genes. Because SRF is not muscle specific, it has been postulated to activate muscle genes by recruiting myogenic accessory factors. Using a bioinformatics-based screen for unknown cardiac-specific genes, we identified a novel and highly potent transcription factor, named myocardin, that is expressed in cardiac and smooth muscle cells. Myocardin belongs to the SAP domain family of nuclear proteins and activates cardiac muscle promoters by associating with SRF. Expression of a dominant negative mutant of myocardin in Xenopus embryos interferes with myocardial cell differentiation. Myocardin is the founding member of a class of muscle transcription factors and provides a mechanism whereby SRF can convey myogenic activity to cardiac muscle genes. PMID- 11439183 TI - Signals transduced by Ca(2+)/calcineurin and NFATc3/c4 pattern the developing vasculature. AB - Vascular development requires an orderly exchange of signals between growing vessels and their supporting tissues, but little is known of the intracellular signaling pathways underlying this communication. We find that mice with disruptions of both NFATc4 and the related NFATc3 genes die around E11 with generalized defects in vessel assembly as well as excessive and disorganized growth of vessels into the neural tube and somites. Since calcineurin is thought to control nuclear localization of NFATc proteins, we introduced a mutation into the calcineurin B gene that prevents phosphatase activation by Ca(2+) signals. These CnB mutant mice exhibit vascular developmental abnormalities similar to the NFATc3/c4 null mice. We show that calcineurin function is transiently required between E7.5 and E8.5. Hence, early calcineurin/NFAT signaling initiates the later cross-talk between vessels and surrounding tissues that pattern the vasculature. PMID- 11439184 TI - Regulation of CLOCK and MOP4 by nuclear hormone receptors in the vasculature: a humoral mechanism to reset a peripheral clock. AB - Circadian clock genes are expressed in the suprachiasmatic nucleus and in peripheral tissues to regulate cyclically physiological processes. Synchronization of peripheral oscillators is thought to involve humoral signals, but the mechanisms by which these are mediated and integrated are poorly understood. We report a hormone-dependent interaction of the nuclear receptors, RAR alpha and RXR alpha, with CLOCK and MOP4. These interactions negatively regulate CLOCK/MOP4:BMAL1-mediated transcriptional activation of clock gene expression in vascular cells. MOP4 exhibits a robust rhythm in the vasculature, and retinoic acid can phase shift Per2 mRNA rhythmicity in vivo and in serum induced smooth muscle cells in vitro, providing a molecular mechanism for hormonal control of clock gene expression. We propose that circadian or periodic availability of nuclear hormones may play a critical role in resetting a peripheral vascular clock. PMID- 11439185 TI - An unfolded putative transmembrane polypeptide, which can lead to endoplasmic reticulum stress, is a substrate of Parkin. AB - A putative G protein-coupled transmembrane polypeptide, named Pael receptor, was identified as an interacting protein with Parkin, a gene product responsible for autosomal recessive juvenile Parkinsonism (AR-JP). When overexpressed in cells, this receptor tends to become unfolded, insoluble, and ubiquitinated in vivo. The insoluble Pael receptor leads to unfolded protein-induced cell death. Parkin specifically ubiquitinates this receptor in the presence of ubiquitin-conjugating enzymes resident in the endoplasmic reticulum and promotes the degradation of insoluble Pael receptor, resulting in suppression of the cell death induced by Pael receptor overexpression. Moreover, the insoluble form of Pael receptor accumulates in the brains of AR-JP patients. Here, we show that the unfolded Pael receptor is a substrate of Parkin, the accumulation of which may cause selective neuronal death in AR-JP. PMID- 11439186 TI - Characterization of a serine protease that cleaves pro-gamma-melanotropin at the adrenal to stimulate growth. AB - The adrenal gland requires stimuli from peptides derived from the ACTH precursor, pro-opiomelanocortin (POMC), to maintain its tonic state. Studies have proposed that a specific postsecretional cleavage of the nonmitogenic N-terminal 16 kDa fragment, also known as pro-gamma-melanotropin (pro-gamma-MSH), is required, releasing shorter fragments that promote adrenal growth. Here, we provide evidence for this hypothesis by the cloning and characterization of a serine protease that is upregulated during growth of the adrenal cortex. It is expressed exclusively in the outer adrenal cortex, the site of cell proliferation, and in the Y1 adrenal cell line. We also show that it is required for growth of Y1 cells, remains bound to the cell surface, and cleaves its substrate, pro-gamma MSH, at a specific bond. PMID- 11439187 TI - Mechanisms contributing to T cell receptor signaling and assembly revealed by the solution structure of an ectodomain fragment of the CD3 epsilon gamma heterodimer. AB - The T cell receptor (TCR) consists of genetically diverse disulfide-linked alpha and beta chains in noncovalent association with the invariant CD3 subunits. CD3 epsilon and CD3 gamma are integral components of both the TCR and pre-TCR. Here, we present the solution structure of a heterodimeric CD3 epsilon gamma ectodomain complex. A unique side-to-side hydrophobic interface between the two C2-set immunoglobulin-like domains and parallel pairing of their respective C-terminal beta strands are revealed. Mutational analysis confirms the importance of the distinctive linkage as well as the membrane proximal stalk motif (RxCxxCxE) for domain-domain association. These biochemical and structural analyses offer insights into the modular pairwise association of CD3 invariant chains. More importantly, the findings suggest how the rigidified CD3 elements participate in TCR-based signal transduction. PMID- 11439188 TI - The DNA polymerase III holoenzyme: an asymmetric dimeric replicative complex with leading and lagging strand polymerases. AB - The DNA Polymerase III holoenzyme forms initiation complexes on primed DNA in an ATP-dependent reaction. We demonstrate that the nonhydrolyzable ATP analog, ATP gamma S, supports the formation of an isolable leading strand complex that loads and replicates the lagging strand only in the presence of ATP, beta, and the single-stranded DNA binding protein. The single endogenous DnaX complex within DNA polymerase III holoenzyme assembles beta onto both the leading and lagging strand polymerases by an ordered mechanism. The dimeric replication complex disassembles in the opposite order from which it assembled. Upon ATP gamma S induced dissociation, the leading strand polymerase is refractory to disassembly allowing cycling to occur exclusively on the lagging strand. These results establish holoenzyme as an intrinsic asymmetric dimer with distinguishable leading and lagging strand polymerases. PMID- 11439189 TI - A coiled-coil from the RNA polymerase beta' subunit allosterically induces selective nontemplate strand binding by sigma(70). AB - For transcription to initiate, RNA polymerase must recognize and melt promoters. Selective binding to the nontemplate strand of the -10 region of the promoter is central to this process. We show that a 48 amino acid (aa) coiled-coil from the beta' subunit (aa 262--309) induces sigma(70) to perform this function almost as efficiently as core RNA polymerase itself. We provide evidence that interaction between the beta' coiled-coil and region 2.2 of sigma(70) promotes an allosteric transition that allows sigma(70) to selectively recognize the nontemplate strand. As the beta' 262--309 peptide can function with the previously crystallized portion of sigma(70), nontemplate recognition can be reconstituted with only 47 kDa, or 1/10 of holoenzyme. PMID- 11439190 TI - Change of plasmid DNA structure, hypermethylation, and Lon-proteolysis as steps in a replicative cascade. AB - I have defined conditions under which RepFIC plasmid DNA can be maintained in a state of lowered helical density. In exponentially growing cultures, the DNA of lowered helical density is present in small amounts but never totally absent, suggesting that it is a normal variant of plasmid maintenance. It is fully methylated at frequent sites by dam-methyltransferase, some not previously recognized, further suggesting that the variant is a precursor of replication. The low-helical density plasmid is present in dam hosts, indicating that methylation is not essential for the change in helical density. The lowered helical density is stabilized in lon hosts, suggesting that Lon-protease may remove both the protein(s) that lower the helical density and the dam methyltransferase after each round of replication. PMID- 11439191 TI - Novel sulfated lymphocyte homing receptors and their control by a Core1 extension beta 1,3-N-acetylglucosaminyltransferase. AB - L-selectin mediates lymphocyte homing by facilitating lymphocyte adhesion to addressins expressed in the high endothelial venules (HEV) of secondary lymphoid organs. Peripheral node addressin recognized by the MECA-79 antibody is apparently part of the L-selectin ligand, but its chemical nature has been undefined. We now identify a sulfated extended core1 mucin-type O-glycan, Gal beta 1-->4(sulfo-->6)GlcNAc beta 1-->3Gal beta 1-->3GalNAc, as the MECA-79 epitope. Molecular cloning of a HEV-expressed core1-beta 1,3-N acetylglucosaminyltransferase (Core1-beta 3GlcNAcT) enabled the construction of the 6-sulfo sialyl Lewis x on extended core1 O-glycans, recapitulating the potent L-selectin-mediated, shear-dependent adhesion observed with novel L-selectin ligands derived from core2 beta1,6-N-acetylglucosaminyltransferase-I null mice. These results identify Core1-beta 3GlcNAcT and its cognate extended core1 O glycans as essential participants in the expression of the MECA-79-positive, HEV specific L-selectin ligands required for lymphocyte homing. PMID- 11439205 TI - HIV DART 2000. Frontiers in drug development for antiretroviral therapies. December 17-21, 2000. Isla Verde, Carolina, Puerto Rico. Abstracts. PMID- 11439206 TI - A randomised trial of accelerated and conventional radiotherapy for stage III and IV squamous carcinoma of the head and neck: a Trans-Tasman Radiation Oncology Group Study. AB - PURPOSE: The aims of this randomized controlled trial were to determine whether there were differences in the disease-free survival (DFS) and toxicity between conventional radiotherapy (CRT) and a continuous 3 week accelerated radiotherapy regimen (ART) in stage III and IV squamous cell carcinoma of the oral cavity, oropharynx, larynx and hypopharynx. PATIENTS AND METHODS: Patients from 14 centres throughout Australia and New Zealand were randomly assigned to either CRT, using a single 2 Gy/day to a dose of 70 Gy in 35 fractions in 49 days or to ART, using 1.8 Gy twice a day to a dose of 59.4 Gy in 33 fractions in 24 days. Treatment allocation was stratified for site and stage. The accrual began in 1991 and the trial was closed in 1998 when the target of 350 patients was reached. RESULTS: The median potential follow-up time was 53 months (range, 14-101). The DFS at 5 years was 41% (95% CI, 33-50%) for ART and 35% (95% CI, 27-43%) for CRT (P=0.323) and the hazard ratio was 0.87 in favour of ART (95% CI, 0.66-1.15). The 5-year disease-specific survival rates were 40% for CRT and 46% for ART (P=0.398) and the loco-regional control was 47% for CRT vs. 52% for ART (P=0.300). The respective hazard ratios were 0.88 (95% CI, 0.65-1.2) and 0.85 (0.62-1.16), favouring the accelerated arm. In the ART arm, confluent mucositis was more severe (94 vs. 71%; P<0.001) and peaked about 3 weeks earlier than in the CRT arm, but healing appeared complete in all cases. There were statistically significant reductions in the probability of grade 2 or greater late soft tissue effects over time in the ART arm (P<0.05), except for the mucous membrane where late effects were similar in both arms. CONCLUSIONS: Differences in DFS, disease specific survival and loco-regional control have not been demonstrated. ART resulted in more acute mucosal toxicity, but this did not result in greater prolongation of the treatment time compared with the CRT arm. There were less late effects in the ART arm, with the exception of late mucosal effects. This trial has confirmed that tumour cell repopulation occurs during conventionally fractionated radiotherapy for head and neck cancer. However, it has also provided additional evidence that overall improvements in the therapeutic ratio using accelerated fractionation strategies are seriously constrained by the need to limit total doses to levels that do not exceed acute mucosal tolerance. The accelerated schedule tested has been shown in this trial to be an acceptable alternative to conventionally fractionated irradiation to 70 Gy. PMID- 11439207 TI - Radiotherapy-related early morbidity in head and neck cancer: quantitative clinical radiobiology as deduced from the CHART trial. AB - BACKGROUND AND PURPOSE: Early reactions after radiotherapy for head and neck cancer may become the limiting factor in current attempts to intensify loco regional therapy through altered fractionation or combination of radiotherapy with chemotherapy. The aim of the present study was to quantify the dependence of early reactions on the dose-fractionation used in radiotherapy and on patients' age and radiation field size. PATIENTS AND METHODS: The data analyzed are from the randomized controlled trial of CHART (continuous hyperfractionated accelerated radiotherapy) vs. conventional radiotherapy in head and neck cancer. The trial accrued 918 patients from March 1990 to April 1995. Several early morbidity items were evaluated weekly for the first 8 weeks after the start of treatment. Weekly evaluation was continued in patients with early reactions extending beyond 8 weeks. Linear regression was used to analyze the time with reactions in individual patients. Polychotomous ordinal response regression was used to analyze the peak grade of early reactions in individual patients. RESULTS: The main findings of this analysis were as follows. (1) The incidence and peak prevalence of confluent mucositis was higher after CHART than after conventional radiotherapy. Therefore, the average time spent with confluent mucositis per patient treated was significantly longer after CHART than after conventional fractionation. (2) In patients who actually developed confluent mucositis, the average duration of this grade of reaction was not significantly different after CHART compared with conventional radiotherapy. (3) Confluent mucositis developed earlier after the start of treatment (2.9 vs. 4.9 weeks) but also started to improve sooner (5.4 vs. 7.5 weeks after the start of treatment) after CHART than after conventional radiotherapy. (4) The dose recovered per 1 day protraction of overall treatment time, D(prolif), was estimated at 0.80 Gy with 95% confidence limits 0.7 and 1.1 Gy/day for human mucosa. (5) For human skin erythema, the estimate of D(prolif) was 0.12 Gy/day with 95% confidence limits -0.12 and 0.22 Gy/day. (6) Highly significant relationships were found between the grade of morphological mucositis on one hand and dysphagia, pain on swallowing and prescribed analgesics on the other. Patients with confluent mucositis had fewer functional problems if this was confined to the larynx as compared with other subsites in the head and neck. (7) Although the incidence of confluent mucositis was higher in the oral cavity and oropharynx than in the hypopharynx and larynx, the radiobiological properties of mucosal reactions did not show significant variation among the various subsites within the head and neck region. (8) For a given dose and overall treatment time, a highly significant increase in incidence and severity of both mucositis and erythema was seen with increasing field size. Thus, a significant dose-volume or dose-area effect exists for both of these tissues. (9) Patients' age had no significant influence on the incidence and severity of mucositis or erythema. CONCLUSIONS: This study provides quantitative estimates of the dose-time and dose-volume relationships for human skin and normal mucosa in the head and neck region based on an analysis of data from 918 patients entered into a randomized-controlled trial of altered dose fractionation in radiotherapy. PMID- 11439208 TI - Time-dose-response relationships in postoperatively irradiated patients with head and neck squamous cell carcinomas. AB - BACKGROUND AND PURPOSE: To define the influence of the dose and time on the response to treatment in postoperatively irradiated head and neck cancer patients and to establish a good prediction of failure. METHODS AND MATERIALS: From January 1985 to December 1995, 214 patients with histologically proven head and neck squamous cell carcinomas were irradiated after radical surgery or single tumour resection according to surgical and histopathological findings. The total doses given ranged between 50 and 75 Gy to the primary bed tumour and between 42 and 56 Gy to the neck with fraction sizes of 1.7-2 Gy/day. The median length of the time interval between surgery and radiotherapy, time of irradiation and total treatment time were 81, 59 and 139 days, respectively. The end-point analyzed was the local-regional tumour control rate at the primary tumour bed and neck for 5 years from the beginning of radiotherapy. Univariate and multivariate analyses were used to determine predictors of failure from among the following studied variables: (i), clinical stage (T/N) of the patients; (ii), tumour grade; (iii), neck surgery; (iv), tumour margins; (v), histological tumour nodal extension; (vi), chemotherapy; (vii), normalized total dose; (viii), time interval between surgery and radiotherapy; (ix), time of irradiation; and (x), total treatment time. RESULTS: The actuarial 5-year tumour control rate for the entire group was 72%, and 92% of the patients who achieved local control are currently alive without disease. Tumour control was inversely related to T stage (83% for T2 vs. 57% for T4) and the probability of local control within each stage was dependent on the N status (> or =71% for T3-T4/N0 vs. 31-44% for T3-T4/N1-N3). Histological N status and tumour margins, but not tumour grade, impacted significantly on tumour control. When local control was analyzed as a function of the dose to the primary, a non-significant negative dose-response relationship was found. The total treatment time was a significant prognostic factor, and the time interval between surgery and irradiation proved to be an independent predictor of failure. CONCLUSIONS: Despite the absence of a statistically significant dose-response relationship, the present results suggest that postoperative irradiation treatment given to patients with head and neck squamous cell carcinomas should not be unduly prolonged, in order to minimize the amount of tumour cell proliferation. In these patients, nodal involvement, positive margins of the resected specimens and time interval between surgery and irradiation were the most important prognostic factors. PMID- 11439209 TI - Combretastatin A-4 and hyperthermia;a potent combination for the treatment of solid tumors. AB - BACKGROUND AND PURPOSE: Attacking tumor vasculature is a promising approach for the treatment of solid tumors. The tubulin inhibitor combretastatin A-4 disodium phosphate (CA-4) is a new vascular targeting drug which displays a low toxicity profile. We wanted to investigate how CA-4 influences tumor perfusion in the BT4An rat glioma and how the vascular targeting properties of CA-4 could be exploited to augment hyperthermic damage towards tumor vasculature. MATERIAL AND METHODS: We used the (86)RbCl extraction technique to assess how CA-4 influences tumor perfusion, and the tumor endothelium was examined for morphological changes induced by the drug. We combined CA-4 (50 mg/kg i.p.) with hyperthermia (44 degrees C, 60 min) at different time intervals to evaluate how therapy should be designed to affect tumor growth, and we studied the tumors histologically to assess tissue viability. RESULTS: We found that CA-4 induced a profound, but transient reduction in tumor perfusion 3-6 h postinjection. If hyperthermia was administered 3-6 h after injecting CA-4, massive hemorrhagic necrosis developed, and tumor response was significantly enhanced compared to simultaneous administration of the two treatment modalities (P<0.005). CA-4 alone had no influence on tumor growth and failed to disrupt the vasculature of the BT4An solid tumors. Interestingly though, a mild endothelial edema was observed in some tumor areas 3 h after injecting CA-4. CONCLUSIONS: We conclude that the combination of CA-4 and hyperthermia is a potent therapeutic option for BT4An tumors, but the selection of adequate time intervals between CA-4 and hyperthermia are imperative to obtain tumor response. PMID- 11439210 TI - Interaction between combretastatin A-4 disodium phosphate and radiation in murine tumors. AB - BACKGROUND AND PURPOSE: The ability of combretastatin A-4 disodium phosphate (CA4DP) to induce vascular damage and enhance the radiation response of murine tumors was investigated. MATERIALS AND METHODS: A C3H mouse mammary carcinoma transplanted in the foot of CDF1 mice and the KHT mouse sarcoma growing in the leg muscle of C3H/HeJ mice were used. CA4DP was dissolved in saline and injected intraperitoneally. Tumor blood perfusion was estimated using 86RbCl extraction and Hoechst 33342 fluorescent labelling. Necrotic fraction was determined from histological sections. Tumors were locally irradiated in non-anaesthetised mice and response assessed by local tumor control for the C3H mammary carcinoma and in vivo/in vitro clonogenic cell survival for the KHT sarcoma. RESULTS: CA4DP decreased tumor blood perfusion and increased necrosis in a dose-dependent fashion in the C3H mammary carcinoma, which was maximal at 250 mg/kg. The decrease in perfusion and induction of necrosis by CA4DP was more extensive in the KHT sarcoma. CA4DP enhanced radiation damage in both tumor types. In the KHT sarcoma this enhancement was independent of whether the drug was given before or after irradiating, whereas for C3H mammary carcinoma the enhancement was only significant when administered at the same time or after the radiation, with no enhancement seen if CA4DP was given before. These effects were drug-dose dependent. CA4DP did not enhance radiation damage in normal skin. CONCLUSIONS: CA4DP enhanced radiation damage in the two tumor models without enhancing normal tissue damage. These radiation effects were clearly consistent with the anti vascular action of CA4DP. PMID- 11439211 TI - Optimisation of radiotherapy for carcinoma of the parotid gland: a comparison of conventional, three-dimensional conformal, and intensity-modulated techniques. AB - BACKGROUND AND PURPOSE: To compare external beam radiotherapy techniques for parotid gland tumours using conventional radiotherapy (RT), three-dimensional conformal radiotherapy (3DCRT), and intensity-modulated radiotherapy (IMRT). To optimise the IMRT techniques, and to produce an IMRT class solution. MATERIALS AND METHODS: The planning target volume (PTV), contra-lateral parotid gland, oral cavity, brain-stem, brain and cochlea were outlined on CT planning scans of six patients with parotid gland tumours. Optimised conventional RT and 3DCRT plans were created and compared with inverse-planned IMRT dose distributions using dose volume histograms. The aim was to reduce the radiation dose to organs at risk and improve the PTV dose distribution. A beam-direction optimisation algorithm was used to improve the dose distribution of the IMRT plans, and a class solution for parotid gland IMRT was investigated. RESULTS: 3DCRT plans produced an equivalent PTV irradiation and reduced the dose to the cochlea, oral cavity, brain, and other normal tissues compared with conventional RT. IMRT further reduced the radiation dose to the cochlea and oral cavity compared with 3DCRT. For nine- and seven-field IMRT techniques, there was an increase in low-dose radiation to non target tissue and the contra-lateral parotid gland. IMRT plans produced using three to five optimised intensity-modulated beam directions maintained the advantages of the more complex IMRT plans, and reduced the contra-lateral parotid gland dose to acceptable levels. Three- and four-field non-coplanar beam arrangements increased the volume of brain irradiated, and increased PTV dose inhomogeneity. A four-field class solution consisting of paired ipsilateral coplanar anterior and posterior oblique beams (15, 45, 145 and 170 degrees from the anterior plane) was developed which maintained the benefits without the complexity of individual patient optimisation. CONCLUSIONS: For patients with parotid gland tumours, reduction in the radiation dose to critical normal tissues was demonstrated with 3DCRT compared with conventional RT. IMRT produced a further reduction in the dose to the cochlea and oral cavity. With nine and seven fields, the dose to the contra-lateral parotid gland was increased, but this was avoided by optimisation of the beam directions. The benefits of IMRT were maintained with three or four fields when the beam angles were optimised, but were also achieved using a four-field class solution. Clinical trials are required to confirm the clinical benefits of these improved dose distributions. PMID- 11439212 TI - Improvements in target coverage and reduced spinal cord irradiation using intensity-modulated radiotherapy (IMRT) in patients with carcinoma of the thyroid gland. AB - BACKGROUND AND PURPOSE: External beam radiotherapy for thyroid carcinoma poses a significant technical challenge as the target volume lies close to or surrounds the spinal cord. The potential of intensity-modulated radiotherapy (IMRT) to improve the dose distributions was investigated. MATERIALS AND METHODS: A planning study was performed on patients with thyroid carcinoma. Plans were generated to irradiate the thyroid bed alone or to treat the thyroid bed and the loco-regional lymph nodes in two phases. Conventional plans with minimal beam shaping were compared to three-dimensional conformal radiotherapy (3DCRT) and inverse-planned IMRT plans to assess target coverage and normal tissue sparing. IMRT techniques were optimized to find the minimum number of equispaced beams required to achieve the clinical benefit and a concomitant boost technique was explored. RESULTS: For the thyroid bed alone and the thyroid bed plus loco regional lymph nodes, conventional and conformal techniques produced low minimum doses to the planning target volume (PTV) if spinal cord tolerance was respected. 3DCRT reduced the irradiated volume of normal tissue (P=0.01). IMRT plans achieved the goal dose to the PTV (P<0.01) and also reduced the spinal cord maximum dose (P<0.01). IMRT, using a concomitant boost technique, produced better target coverage than a two-phase technique. For both the two-phase and concomitant boost techniques, IMRT plans with seven and five equispaced fields produced similar dose distributions to nine fields, but three fields were significantly worse. CONCLUSIONS: 3DCRT reduced normal tissue irradiation compared to conventional techniques, but did not improve PTV or spinal cord doses. IMRT improved the PTV coverage and reduced the spinal cord dose. A simultaneous integrated boost technique with five equispaced fields produced the best dose distribution. IMRT should reduce the risk of myelopathy or may allow dose escalation in patients with thyroid cancer. PMID- 11439213 TI - Pre-treatment dosimetric verification by means of a liquid-filled electronic portal imaging device during dynamic delivery of intensity modulated treatment fields. AB - BACKGROUND AND PURPOSE: Although intensity modulated radiation therapy is characterized by three-dimensional dose distributions which are often superior to those obtained with conventional treatment plans, its routine clinical implementation is partially held back by the complexity of the beam verification. This is even more so when a dynamic multileaf collimator (dMLC) is used instead of a segmented beam delivery. We have therefore investigated the possibility of using a commercially available, liquid-filled electronic portal imaging device (EPID) for the pre-treatment quality assurance of dynamically delivered dose distributions. METHODS AND MATERIALS: A special acquisition mode was developed to optimize the image acquisition speed for dosimetry with the liquid-filled EPID. We investigated the accuracy of this mode for 6 and 18 MV photon beams through comparison with film and ion chamber measurements. The impact of leaf speed and pulse rate fluctuations was quantified by means of dMLC plans especially designed for this purpose. Other factors influencing the accuracy of the dosimetry (e.g. the need for build-up, remanence of the ion concentration in the liquid and bulging of the liquid at non-zero gantry angles) were studied as well. We finally compared dosimetric EPID images with the corresponding image prediction delivered without a patient in the beam. RESULTS: The dosimetric accuracy of the measured dose distribution is approximately 2% with respect to film and ion chamber measurements. The accuracy declines when leaf speed is increased beyond 2 cm/s, but is fairly insensitive to accelerator pulse rate fluctuations. The memory effect is found to be of no clinical relevance. When comparing the acquired and expected distributions, an overall agreement of 3% can be obtained, except at areas of steep dose gradients where slight positional shifts are translated into large errors. CONCLUSIONS: Accurate dosimetric images of intensity modulated beam profiles delivered with a dMLC can be obtained with a commercially available, liquid-filled EPID. The developed acquisition mode is especially suited for fast and accurate pre-treatment verification of the intensity modulated fields. PMID- 11439214 TI - Tolerances for the accuracy of photon beam dose calculations of treatment planning systems. AB - BACKGROUND AND PURPOSE: To design a consistent set of criteria for acceptability of photon beam dose calculations of treatment planning systems. The set should be applicable in combination with a test package used for evaluation of a treatment planning system, such as the ones proposed by the AAPM Task Group 23 or by the Netherlands Commission on Radiation Dosimetry. RESULTS: Tolerances have been defined for the accuracy with which a treatment planning system should be able to calculate the dose in different parts of a photon beam: the central beam axis and regions with large and small dose gradients. For increasing complexity of the geometry, wider tolerances are allowed, varying between 2 and 5%. For the evaluation of a large number of data points an additional quantity, the confidence limit, has been introduced, which combines the influence of systematic and random deviations. The proposed tolerances have been compared with other recommendations for a number of clinically relevant examples, showing considerable differences, which are partly due to the way the complexity of the geometry is taken into account. Furthermore differences occur if criteria for acceptability of dose calculations are related either to the local dose value or to a normalized dose value. CONCLUSIONS: Although it is acknowledged that the general aim must be to have good agreement between dose calculation and the actual dose value, e.g. within 2% or 2 mm, current day algorithms and their implementation into commercial treatment planning systems result often in larger deviations. A high accuracy can at present only be achieved in relatively simple cases. The new set of tolerances and the quantity confidence limit have proven to be useful tools for the acceptance of photon beam dose calculation algorithms of treatment planning systems. PMID- 11439215 TI - Application of a test package in an intercomparison of the photon dose calculation performance of treatment planning systems used in a clinical setting. AB - BACKGROUND AND PURPOSE: Testing the performance of treatment planning systems by using the AAPM Task Group 23 test package is a useful approach, but has its limitations. To be able to include technical developments, such as the asymmetric collimator, it was decided to remeasure the AAPM data set on more modern radiotherapy equipment, to extend the test geometries, and to evaluate the use of the new package. MATERIALS AND METHODS: A coherent set of beam data of 6, 10 and 18 MV photon beams was measured on two modern linear accelerators. These data served as input data in seven commercially available treatment planning systems, which were clinically in use in different radiotherapy departments. Next, a test package was measured which included a missing tissue geometry and fields with asymmetrical collimator setting, with and without a wedge. RESULTS: The absolute dose prediction from the different treatment planning systems in which the measured beam data were entered, was compared for all test points with the results of direct measurements. The criteria of acceptability were exceeded by some systems in cases of irregular field geometry and missing tissue geometry. The majority of the systems had difficulties with accurate dose calculation for asymmetrically wedged fields. CONCLUSIONS: The application of the new test package did not introduce insuperable difficulties and was highly appreciated by the participating centres. Most systems performed reasonably well for the majority of the beam geometries, with the exception of asymmetrically wedged beams. The extended test package is available for other users or user groups for the purpose of commissioning new treatment planning systems, or new releases of existing systems. PMID- 11439216 TI - Commissioning of a commercially available system for intensity-modulated radiotherapy dose delivery with dynamic multileaf collimation. AB - PURPOSE: To commission commercially available equipment for intensity-modulated radiotherapy (IMRT) using dynamic multileaf collimation (DMLC). MATERIALS AND METHODS: First, the stability in leaf positioning and in realized IMRT profiles on a Varian 2300 C/D machine were determined as a function of time and gantry angle, and as a result of treatment interrupts. Second, dose distributions calculated with the CadPlan (Varian) treatment planning system, using leaf trajectories calculated with the leaf motion calculator (LMC) algorithm, were compared with distributions realized at the 2300 C/D unit. RESULTS: Day-to-day and gantry angle variations in leaf positioning and dose delivery were very small (less than 0.1-0.2 mm and 2%). The effect of treatment interrupts on measured dose distributions was less than 2%. The agreement between the final dose distribution calculated by CadPlan and the measured dose was generally within 2%, or 2 mm at steep dose gradients, using a leaf transmission value of 1.8% and a leaf separation value of 2 mm in LMC. For narrow peaks, deviations of up to 6% were observed. LMC does not synchronize adjacent leaf trajectories resulting in tongue-and-groove underdosages of up to 29% for extreme cases. CONCLUSIONS: The 2300 C/D machine is suitable for accurate and reproducible DMLC treatments. The agreement between dose predictions with LMC and CadPlan, and realized doses at this unit is clinically acceptable for most cases. However, differences between calculated and actual dose values may exist in peaked fluences or due to tongue and-groove effects. Therefore, pretreatment dosimetric verification for each patient is recommended. PMID- 11439218 TI - Tissue specific changes in the expression of glutamate-cysteine ligase mRNAs in mice exposed to methylmercury. AB - Glutamate-cysteine ligase (GLCL), the rate-limiting enzyme in glutathione (GSH) synthesis is composed of two subunits, a catalytic (GLCLc) and a regulatory subunit (GLCLr). These two subunits are known to be differentially regulated in vitro, in different cell types and in response to various xenobiotic exposures. In this study, we examined whether these two subunits can also be differentially regulated in vivo. We found that GLCLc and GLCLr are differentially regulated at the transcriptional level in a tissue-dependent manner in female mice treated with methylmercury (MeHg). MeHg caused a downregulation of both subunit mRNAs in the liver, upregulation of both subunit mRNAs in the kidney and upregulation of only the catalytic subunit mRNA in the small intestine of female mice treated with a single dose of MeHg (6 mg/kg) by intraperitoneal injection. These results suggest that GLCLc and GLCLr can be differentially regulated in vivo, and that this regulation is tissue dependent in the mouse. PMID- 11439219 TI - Phenobarbitone-induced liver response in wild type and in p53 deficient mice. AB - The tumour suppressor protein, p53, is involved in the regulation of apoptosis and growth arrest following DNA damage. Mutations of the p53 gene are found in 50 55% of all human cancers (Hollstein et al. Nucl. Acid Res. 22 (1994) 3551), including hepatocellular carcinomas. Phenobarbitone (PB) is a non-genotoxic hepatocarcinogen in rats and mice. With commercial availability of mice where one or both alleles of p53 have been removed we have examined the effect of PB in wild type C57BL/6J mice (p53 +/+), and p53 deficient mice (+/- and -/- p53) to determine whether p53 plays a role in the PB induced liver response. In each strain of mice, chronic administration caused liver enlargement, which was associated with centrilobular hepatocyte hypertrophy and a transient hyperplasia. In addition, an increase in centrilobular epidermal growth factor receptor and its ligand, transforming growth factor alpha and a decrease in mannose-6 phosphate receptor and its mitoinhibitory ligand, TGFbeta1 was also observed immunohistochemically. The similar response in all three strains indicates that p53 probably plays no role in the early PB induced liver effects of hypertrophy and changes in growth factor expression. PMID- 11439220 TI - Effect of fructose-1, 6-diphosphate versus diphenhydramine on mortality in compound 48/80-induced shock. AB - Fructose-1,6-diphosphate (FDP) has a salutary effect on hemorrhagic, traumatic and endotoxic shock. The role of FDP on compound 48/80-induced shock was therefore investigated. Sprague Dawley aged male rats (448+/-7.4 gm body weight) were randomly assigned into three groups and treated intraperitoneally with diphenhydramine (DPHM) 15 mg/kg (n=11), 12.5 ml of 10% FDP (n=10) and 12.5 ml saline (n=10). The rats were injected with compound 48/80 (5 mg/kg) 30 min later, and monitored every 10 min for 60 min. Arterial pressure was higher in FDP rats than in DPHM (P<0.01) or saline (P<0.005) groups. Plasma potassium (K(+)) was lower in the FDP group (P<0.01). Arterial pO2 and pCO2 were within physiological range in all groups. A profound decrease in arterial pH and bicarbonate (HCO3(-)) was also observed in all groups. Mortality at 48 h in the saline group was 100%, in the DPHM group 91%, and in the FDP group 20% (P<0.001 and P<0.005, respectively). FDP improved survival significantly in this study. PMID- 11439221 TI - Development of preimplantation mouse embryos after exposure to a 50 Hz magnetic field in vitro. AB - Effect of sinusoidal 50 Hz magnetic field (MF) on development of preimplantation CBA/S mouse embryos in vitro was studied. Superovulated and in vivo fertilized preimplantation embryos were collected at one cell stage and divided to control and MF-exposed groups. Sinusoidal 50 Hz MF with field strength of 10 A/m r.m.s., corresponding a flux density of 13 microT r.m.s., was used to expose the embryos in culture at 37 degrees C in a CO2-incubator. The developmental stage and abnormalities were recorded twice daily except once daily during weekends. The vitality and developmental stages of the embryos were similar in both groups although slightly more dead embryos were found during the 1st day in MF-exposed group (P<0.05) and the development of MF-exposed embryos was slightly impaired. In conclusion, the exposure to sinusoidal 50 Hz MF at field strength of 10 A/m did not significantly disturb the development of the mouse embryos in vitro up to the blastocyst stage. PMID- 11439222 TI - Oxidative stress and liver toxicity in rats and human hepatoma cell line induced by pentachlorophenol and its major metabolite tetrachlorohydroquinone. AB - Pentachlorophenol (PCP) is a pesticide used worldwide in industrial and domestic applications. It is used extensively as biocide and wood preservatives. Metabolic studies carried out in rodents and human liver homogenates have indicated that PCP undergoes oxidative dechlorination to form tetrachlorohydroquinone (TCHQ). Free radical catalyzed tissue injury is thought to play a fundamental role in human disease. In the present study, we examined the effects of PCP and TCHQ on the induction of lipid peroxidation and liver injury in rats. In addition, the cytotoxic dose, cell death mechanisms and related gene expressions induced by PCP and TCHQ were also determined for human hepatoma cell line (Hep G2). The results indicated that more toxic effects could be observed both in rats and human hepatoma cell line treated with TCHQ than its parent compound, PCP. Oxygen species may be involved in the mechanism of TCHQ intoxication since the urinary 8 epi-PGF2alpha and AST, ALT activities can be induced by TCHQ and attenuated by vitamin E treatment. Apoptosis features were found in cells treated with TCHQ but not PCP. TCHQ-induced cell damage may issue signals for the induction of HSPs, the decrease of the bcl/bax protein ratio and the decrease of CAS gene, whereas the PCP-induced damage may not. PMID- 11439223 TI - DMT1 gene expression and cadmium absorption in human absorptive enterocytes. AB - Divalent metal transporter 1 (DMT1) is a transmembrane, proton-coupled metal ion transporter that is upregulated in the duodenum of iron-deficient rodents and in hereditary hemochromatosis patients, suggesting that it may constitute a key factor in the uptake of dietary iron. Functional expression studies in Xenopus oocytes have shown that DMT1 not only mediates transport of iron but also other divalent metal ions, including the toxic metal cadmium. In the present study, the correlation between the cadmium absorption process and gene expression of DMT1 was investigated in an experimental model of human absorptive enterocytes. Fully differentiated Caco-2 cells were grown in monolayers and treated with iron supplemented medium or control medium for 1, 3 or 7 days. At each time point, cadmium transport experiments across the Caco-2 cell monolayers were performed and gene expression of DMT1 measured. Iron treatment for 3 and 7 days resulted in a 50% reduction in the cadmium uptake and a 75% reduction in the transport of the metal across the basolateral membrane. No effects were observed at 24 h. Gene expression of DMT1 in the iron-treated Caco-2 cells was reduced by about 50% at 3 and 7 days and thus, correlated well with the uptake of cadmium. In summary, our results indicate that the uptake of cadmium into human absorptive enterocytes may be mediated by DMT1. PMID- 11439224 TI - Mycotoxin detoxication of animal feed by different adsorbents. AB - The contamination of animal feed with mycotoxins represents a worldwide problem for farmers. These toxins originate from molds whose growth on living and stored plants is almost unavoidable particularly under moist conditions. Mycotoxin containing feed can cause serious diseases in farm animals resulting in suffering and even death and thus can cause substantial economic losses. The most applied method for protecting animals against mycotoxicosis is the utilization of adsorbents mixed with the feed which are supposed to bind the mycotoxins efficiently in the gastro-intestinal tract. Aluminosilicates are the preferred adsorbents, followed by activated charcoal and special polymers. The efficiency of mycotoxin binders, however, differs considerably depending mainly on the chemical structure of both the adsorbent and the toxin. This review describes the most important types of adsorbents and the respective mechanisms of adsorption. Data of the in vitro and in vivo efficacy of detoxication are given. PMID- 11439225 TI - Edovascular grafting 'state of the art'. PMID- 11439226 TI - Imaging of abdominal aortic aneurysm before and after endoluminal stent-graft repair. AB - Imaging is an essential component of endoluminal aneurysm repair. Detailed imaging with computed tomography, magnetic resonance imaging and angiography, alone or in combination, is required for the initial assessment and planning. Careful, lifelong follow-up is essential since complications of endoluminal repair may take months or years to appear. Follow-up imaging requires a combination of plain film radiography, colour Doppler ultrasound and helical computed tomography. Magnetic resonance imaging may be valuable for the follow-up of non ferro-magnetic endografts and intra-arterial angiography will be required for specific cases. PMID- 11439227 TI - Endoluminal repair of abdominal aortic aneurysms - state of the art. AB - This paper considers the historical aspects of endovascular aneursym repair; the major findings of our Departments experience over an 812-year period and reviews recent developments in endovascular prostheses. Analysis of 400 patients undergoing primary repair of abdominal aortic aneurysm between 1992 and 2000 revealed a perioperative mortality rate of 2.7% and primary conversion rate of 5%. With sequential studies it was shown that the outcome was better with bifurcated/aorto uni iliac grafts than tube grafts; better with second generation prostheses than first generation protheses and that survival in consecutive patients treated concurrently by open repair and endoluminal repair was superior in the endoluminal group. Endoluminal AAA repair is at a critical point of its development. It is unquestioned that it can dramatically reduce the need for intensive care and length of hospital stay and more recently it has been reported that survival is improved compared with open repair. The need for lifetime surveillance, the probability of graft failure and need for re-intervention, however, negate some of the advantages. The small incidence of unpredictable rupture following endoluminal AAA repair is a timely reminder of the need for continued careful follow-up. PMID- 11439228 TI - Complications in endoluminal repair of abdominal aortic aneurysms. AB - In this review the technical and systemic complications occurring during endoluminal repair of abdominal aortic aneurysms are summarized. According to the data in the literature the technical success-rate of the endoluminal procedure should be >90% and the overall complication-rate <10%. It should be differentiated between complications which have an adverse effect on the outcome of the procedure, and technical problems, which complicate the procedure but do not affect the outcome. The majority of the technical problems can be solved endoluminally. The 30-day mortality rate should be in the same range as elective open surgical repair. One major factor influencing the immediate results is the experience of the interventional team. PMID- 11439229 TI - Lessons learnt from the EUROSTAR registry on endovascular repair of abdominal aortic aneurysm repair. AB - OBJECTIVE: The EUROSTAR project is a multicentred database of the outcome of endovascular repair of infra-renal aortic aneurysms. To date 92 European centres of vascular surgery have contributed. The purpose of the article here is to review the medium term (up to 4 years) results of endovascular aneurysm repair as reported to Eurostar. PATIENTS AND METHODS: Patients intended for endovascular aneurysm repair were notified to the EUROSTAR Data Registry Centre before treatment in order to eliminate bias due to selective reporting. The following data was collected on all patients: (1) their demographic details and the anatomical characteristics of their aneurysms, (2) details of the endovascular device used, (3) procedural complications and the immediate outcome, (4) results of contrast enhanced CT imaging at 3, 6, 12 and 18 months after operation and at yearly intervals thereafter, (5) all adverse events. Life table analysis was performed to determine the cumulative rates of: (1) death from all causes, (2) secondary intervention. Risk factors for rupture and late conversion were identified by regression analysis. RESULTS: By July 2000, 2862 patients had been registered and their median duration of follow-up was 12 mo (range 0-72). Successful deployment was achieved in 2812 patients with a perioperative (30 day) mortality of 2.9%. In 2464 patients enrolled by March 2000 late rupture of the aneurysm occurred in 14 patients for an annual cumulative rate of 1%. The significant factors were proximal type I endoleak (P=0.001), midgraft (type III) endoleak (P=0.001), graft migration (P=0.001) and post-operative kinking of the endograft (P=0.001). Forty-one patients had late conversion to open repair for an annual cumulative rate (risk) of approximately 2.1%. Risk factors (indications) for late conversion were: proximal type I endoleak (P=0.001), midgraft (type III) endoleak (P=0.001), type II endoleak (P=0.003), graft migration (P=0.001), graft kinking (P=0.001) and distal type I endoleak (P=0.001). CONCLUSIONS: Endovascular repair of infra-renal aortic aneurysms using the first and second-generation devices that predominated in this study was associated with a risk of late failure of 3% per year, based upon an analysis of observed primary endpoints of rupture and conversion. Eurostar continues to provide responsible evaluation of the technique for the benefit of both physicians and the industry. PMID- 11439230 TI - Endovascular stent-graft management of thoracic aortic diseases. AB - The traditional standard therapy for descending thoracic aortic aneurysm (TAA) is open operative repair with graft replacement of the diseased aortic segment. Despite important advances in surgical techniques, anesthetic management, and post-operative care over the last 30 years, the mortality and morbidity of surgery remains considerable, especially in patients at high risk for thoracotomy because of coexisting severe cardiopulmonary abnormalities or other medical diseases. The advent of endovascular stent-graft technology provides an alternative to open surgery for selected patients with TAA. The initial experience suggests that stent-graft therapy potentially may reduce the operative risk, hospital stay and procedural expenses of TAA repair. These potential benefits are especially attractive for patients at high risk for open TAA repair. Current results of endovascular TAA therapy document operative mortalities of between 0 and 4%, aneurysm thrombosis in 90 and 100% of cases, and paraplegia as a complication in 0 and 1.6% of patients. The early success of stent-graft repair of TAA has fostered the application of these devices for the management of a wide variety of thoracic aortic pathologies, including acute and chronic dissection, intramural hematoma, penetrating ulcer, traumatic injuries, and other diseases. The results of prospective controlled trials that compare the outcomes of stent graft therapy with those of surgical treatment in patients with specific types of aortic disease are anxiously awaited before recommendations regarding the general use of these new devices can be made with confidence. PMID- 11439231 TI - Screening women at high risk of breast cancer on the basis of evidence. AB - Geneticists are able to identify the risk of breast cancer. Strategies on offer include prevention, early diagnosis by screening, and prophylactic surgery. This paper analyses the evidence for offering screening. The radiation dose of mammography has been measured, but the risk is not fully known. Mammography screening of women of 40-50 years in the normal population has known effect. Little evidence is available for women under 40 years or for women with genetic susceptibility to breast cancer. Dense parenchymal pattern is associated with high grade cancers, and is both a risk factor and a reason for impaired screening sensitivity. Whether this applies to younger women or women at high risk is speculative. The pathological features of the cancers in gene carriers show differences from those occurring in normal women. This work should be correlated with imaging features. There is no literature to support the use of newer imaging methods in these women. Ultrasound and MRI avoid radiation and may be useful in dense breasts. SestaMIBI and PET scanning are not yet mature enough for screening, and may never have such a role. Any newer modality must be subjected to a formal randomised trial before being offered to screen women at high risk. PMID- 11439232 TI - Use of artificial neural networks (computer analysis) in the diagnosis of microcalcifications on mammography. AB - INTRODUCTION/OBJECTIVE: The purpose of this study was to evaluate a computer based method for differentiating malignant from benign clustered microcalcifications, comparing it with the performance of three physicians. METHODS AND MATERIAL: Materials for the study are 240 suspicious microcalcifications on mammograms from 220 female patients who underwent breast biopsy, following hook wire localization under mammographic guidance. The histologic findings were malignant in 108 cases (45%) and benign in 132 cases (55%). Those clusters were analyzed by a computer program and eight features of the calcifications (density, number, area, brightness, diameter average, distance average, proximity average, perimeter compacity average) were quantitatively estimated by a specific artificial neural network. Human input was limited to initial identification of the calcifications. Three physicians-observers were also evaluated for the malignant or benign nature of the clustered microcalcifications. RESULTS: The performance of the artificial network was evaluated by receiver operating characteristics (ROC) curves. ROC curves were also generated for the performance of each observer and for the three observers as a group. The ROC curves for the computer and for the physicians were compared and the results are:area under the curve (AUC) value for computer is 0.937, for physician-1 is 0.746, for physician-2 is 0.785, for physician-3 is 0.835 and for physicians as a group is 0.810. The results of the Student's t-test for paired data showed statistically significant difference between the artificial neural network and the physicians' performance, independently and as a group. DISCUSSION AND CONCLUSION: Our study showed that computer analysis achieves statistically significantly better performance than that of physicians in the classification of malignant and benign calcifications. This method, after further evaluation and improvement, may help radiologists and breast surgeons in better predictive estimation of suspicious clustered microcalcifications and reduce the number of biopsies for non-palpable benign lesions. PMID- 11439233 TI - Adult adrenal haemorrhage: an unrecognised complication of renal vein thrombosis. AB - There are many predisposing factors for neonatal adrenal haemorrhage but the causative factors are different in adults. Several cases of neonatal adrenal haemorrhage have been reported in association with renal vein thrombosis. This complication has not been documented in the adults. The presence of an adrenal mass in the setting of renal vein thrombosis should raise the possibility of adrenal haemorrhage even though this is extremely uncommon in adults. PMID- 11439234 TI - Using biological indices to classify schizophrenia and other psychotic patients. AB - Although classification of mental disorders using more than clinical description would be desirable, there is scant evidence that available laboratory tests (i.e. biological indices) would provide more valid classifications than current diagnostic systems (e.g. DSM-IV). We used cluster analysis of four biological variables to classify 163 psychotic patients and 83 nonpsychiatric comparison subjects. Analyses revealed a three-cluster solution with the first cluster reflecting electrodermal deviance, the second cluster representing nondeviant biological function, and the third cluster reflecting increased nailfold plexus visibility and ocular motor dysfunction. To assess the construct validity of proband clusters we examined ocular motor performance in 156 first-degree relatives as a function of proband cluster membership. First-degree relatives of third cluster probands exhibited worse ocular motor performance than relatives of other cluster probands. Additionally, better classification sensitivity and specificity were obtained for the relatives when they were grouped by proband cluster than by proband DSM-IV diagnosis. When a single proband characteristic (i.e. eyetracking performance) was used to group relatives, classification sensitivity and specificity failed to significantly increase over grouping by proband DSM-IV diagnosis. Multivariate biologically defined clusters may offer an advantage over DSM-IV classification when examining nosology and etiology of psychotic disorders. PMID- 11439235 TI - Is reduced dermatoglyphic a-b ridge count a reliable marker of developmental impairment in schizophrenia? AB - BACKGROUND: Finger and hand prints are formed during the late first and second trimester of foetal development, after which they remain unchanged. Their expression may be influenced by both genetic and environmental factors. Some studies have suggested that a reduced total finger ridge count (TFRC) and, in particular, a reduce total a-b ridge count (TABRC), may be associated with schizophrenia. AIM: To study these two variables in a large, ethnically homogenous sample and to compare our findings with those of other recent studies. METHOD: Finger and hand prints of 150 people with DSM-III-R schizophrenia were compared with those of 92 healthy controls. RESULTS: Patients had a reduced mean TABRC (P = 0.03) compared with controls. There was a significant (P=0.02) linear trend for lower TABRC and increasing incidence of schizophrenia (ORlineartrend = 1.3; 95%CI1.1-1.7), implying a continuous increase in the risk for schizophrenia with reduction in TABRC. No significant difference between groups was observed for TFRC. CONCLUSION: These results provide further evidence that dermatoglyphic abnormalities exist in at least some patients with schizophrenia and that the a-b ridge count may be a marker of disruption, probably environmental, that occurs when the developing brain may also be particularly vulnerable to such insult. These findings support the concept that some cases of schizophrenia may be due to adverse intrauterine events. PMID- 11439236 TI - Dermatoglyphic fluctuating asymmetry and atypical handedness in schizophrenia. AB - Atypical handedness and dermatoglyphic abnormalities are hypothesized to reflect a neurodevelopmental disturbance in schizophrenia. Developmental instability, indexed by dermatoglyphic fluctuating asymmetry (FA), reflects the degree to which an individual's ontogenetic program is maintained and provides a useful framework in which to consider atypical handedness in schizophrenia. Thirty patients diagnosed with schizophrenia were compared with 37 matched healthy controls on levels of dermatoglyphic FA, a demonstration task determining hand preference and a test of relative hand skill. Multivariate analyses established that patients demonstrated greater FA and more atypical hand skill compared with controls. In patients, but not in controls, there was a strong positive association between a measure of FA and a measure of atypical hand skill, suggesting that these markers of neurodevelopmental disturbance are related in schizophrenia. On a measure of hand preference, patients were more likely than controls to be classified as mixed handed than either right or left handed. Results from the present study support the conjecture of greater developmental instability in schizophrenia affecting neurodevelopmental processes, including those conferring manual dominance. PMID- 11439237 TI - Neuroleptics normalize increased release of interleukin- 1 beta and tumor necrosis factor-alpha from monocytes in schizophrenia. AB - Some recent reports show that schizophrenia is accompanied by changes in lymphocyte activity. This study investigated the activity of monocytes by determining their release of interleukin- 1 beta (IL- 1 beta) and tumor necrosis factor-alpha (TNF-alpha). Monocytes were immunomagnetically isolated from the peripheral blood of schizophrenic patients before and after neuroleptic medication and stimulated by lipopolisaccharide (LPS) in vitro. The monocytes of schizophrenic patients released significantly higher amounts of IL- 1 beta and TNF-alpha than those of healthy controls. Treatment with the typical neuroleptics haloperidol and perazine decreased the release of IL- 1 beta and TNF-alpha to the control levels. The study has shown that the activity of monocytes is increased in schizophrenia and that neuroleptic treatment normalizes this activity. PMID- 11439238 TI - Low platelet count in a 22q11 deletion syndrome subtype of schizophrenia. AB - BACKGROUND: 22q11 Deletion Syndrome (22qDS) is a genetic syndrome associated with various physical features and schizophrenia. Some reports have identified thrombocytopenia (platelet count < 150 x 10(9)/l) in individuals with 22qDS, especially children. We investigated whether adults with 22qDS and schizophrenia (22qDS-SZ) have lower platelet counts than other patients with schizophrenia (SZ). METHOD: Complete blood counts (CBC) were recorded from medical records for 18 22qDS-SZ and 60 SZ subjects. Five CBCs per subject were randomly selected and used to calculate a within-subject mean for analyses. RESULTS: 22qDS-SZ subjects had significantly lower mean platelet counts than comparison SZ subjects (142.2 x 10(9)/l versus 282.5 x 10(9)/l, t = -11.5, p < 0.0001). Ten 22qDS-SZ (55%) and no comparison subjects had thrombocytopenia. CONCLUSIONS: These results suggest that thrombocytopenia may be a common feature of 22qDS and that low platelet counts may comprise a readily available screening criterion to help identify this genetic syndrome among adults with schizophrenia. PMID- 11439239 TI - Schizotypy and the shift from dextrality: a study of handedness in a large non clinical sample. AB - Numerous previous studies have reported associations between schizophrenia/schizotypy and atypical handedness. While there is broad agreement that schizophrenia/schizotypy is associated with a shift away from typical (strong right) handedness, it is not entirely clear what aspect of atypical handedness is implicated: 'ambiguous' handedness (the absence of hand preference for given actions) or 'mixed handedness' (different hand preferences for different actions). The present study used several indices of handedness (derived from the Annett scale) to address these questions in 3000 + (mostly) University student subjects recruited by electronic mail. It was found that positive schizotypy (indexed by a scale of 'unusual experiences') was associated with both aspects of atypical handedness. These findings are discussed in the light of various possible explanations of the association between handedness and schizophrenia/schizotypy. PMID- 11439240 TI - An animal model of nicotinic-acid-induced vasodilation: effect of haloperidol, caffeine and nicotine upon nicotinic acid response. AB - BACKGROUND: The normal vasodilatory response to ingestion of nicotinic acid (NA) is impaired in some patients with schizophrenia. It is unclear whether the impairment is a feature of the disorder itself or to a confounding factor such as neuroleptics, caffeine or nicotine use. METHODS: To address this question in a controlled manner, we have developed an animal (rat) model of NA-induced vasodilation, in which response is monitored by measuring change in skin temperature. RESULTS: We observed that (i) acute administration of acetylsalicylic acid (100mg/kg), caffeine (2.5mg/kg) and haloperidol (0.1 or 0.5mg/kg) and (ii) chronic administration of haloperidol (0.2mg/kg/day) significantly inhibited NA (30 mg/kg) response, whereas neither acute (0.25mg/kg) or chronic (0.5mg/kg/day for 14 days) administration of nicotine, or chronic administration of caffeine (5mg/kg/day for 14 days) had any significant effect upon NA response. CONCLUSIONS: Our data suggest that at least one drug commonly used to treat schizophrenia (haloperidol) can interfere with the vasodilatory response to NA. Studies using non-medicated patients with schizophrenia are required to determine whether reduced vasodilatory response to NA in schizophrenia is a feature of the disorder or a consequence of treatment. PMID- 11439241 TI - A comparison of memory for verbal and non-verbal material in schizophrenia. AB - Verbal memory impairment has been well explored in schizophrenia, but it is unclear whether findings relate to the type of material to be learned or the component process required by the memory task. Also, sparse data on non-verbal memory also open the question of how well schizophrenia patients encode this material. We tested whether episodic memory performance in schizophrenia varies as a function of stimulus material (verbal/non-verbal) and determined the integrity of various component memory processes. Memory tests that differ in stimulus material (words, California Verbal Learning Test, CVLT; designs, Biber Figure Learning Test-Extended, BFLT-E) yet produce similar memory component measures were used. Subjects were 28 neuroleptic-medicated inpatients with a diagnosis of chronic schizophrenia. Results showed that both verbal and non verbal memory performance was impaired relative to age-matched controls. Learning and recall measures were most severely impaired, with memory storage problems and impairment in recognition memory evident. On the verbal task, the relative sparing of recognition memory suggested retrieval processes, in addition to encoding processes, were disrupted. On the non-verbal task, the deficits appeared more limited to encoding. Therefore, while the operational integrity of components such as encoding were compromised regardless of material, retrieval processes showed material-specific effects. To the degree verbal and non-verbal memory functions can be lateralized in the brain, these data support the possibility of deficits in both right and left hemisphere declarative memory systems in schizophrenia. PMID- 11439242 TI - Effective risperidone treatment for simple deteriorative disorder (simple schizophrenia): a case report. PMID- 11439243 TI - The Drosophila cell shape regulator c-Jun N-terminal kinase also functions as a stress-activated protein kinase. AB - Mammalian c-Jun N-terminal kinases (JNKs) are members of a group of stress activated intracellular signalling molecules within the MAP kinase family. Molecular genetic analysis of a highly evolutionarily conserved Drosophila JNK homologue, DJNK, has demonstrated that this molecule plays an essential developmental role in cell shape regulation. However, it remains to be determined whether DJNK also responds to the broad range of cellular stresses and other stimuli that affect its mammalian counterpart. Here we demonstrate that c-Jun, a substrate for mammalian JNKs, is a specific substrate for DJNK and that an antiserum that cross-reacts with activated mammalian JNK at the conserved threonyl-prolyl-tyrosyl (TPY) motif within the activation loop also specifically recognises the activated form of DJNK. Using these two assays, we show that DJNK activity is stimulated in cultured cells by several treatments that activate mammalian JNKs, including addition of arsenite, vanadate and ceramide derivatives. It is therefore concluded that in addition to its essential developmental functions, DJNK plays an important role in stress responses that mirrors its mammalian counterpart. PMID- 11439244 TI - Mode of action of Bacillus thuringiensis PS86Q3 strain in hymenopteran forest pests. AB - The mode of action of Cry toxins has been described principally in lepidopteran insects as a multistep process. In this work we describe the mode of action of a Cry toxin active in the common pine sawfly Diprion pini (Hymenoptera, Diprionidae), considered a major forest pest in Europe. Strain PS86Q3 contains a long bipyramidal crystal composed of five major proteins. The N-terminal sequence shows that the 155 kDa protein corresponds to Cry5B toxin and the other proteins belong to the Cry5A subgroup. PCR analysis indicates the presence of cry5Ac and cry5Ba genes, suggesting that Cry5A protein should be Cry5Ac. Activation of protoxins with trypsin or with midgut content from D. pini and Cephacia abietis (Hymenoptera, Pamphiliidae) (spruce webspinning sawfly), another important hymenopteran forest pest, produced a single 75 kDa toxin that corresponded to Cry5A by N-terminal sequence and is responsible for the insecticidal activity. Homologous competition experiments with D. pini and C. abietis brush border membrane vesicles (BBMV) showed that the binding interaction of Cry5A is specific. Membrane potential measurements using a fluorescent dye indicate that Cry5A toxin at nM concentration caused immediate permeability changes in the BBMV isolated from both hymenopteran larvae. The initial response and the sustained permeability change are cationic as previously shown for Cry1 toxins. These results indicate that the hymenopteran specific Cry5A toxin exerts toxicity by a similar mechanism as Cry1 toxins. PMID- 11439245 TI - Identification of a defensin from the hemolymph of the American dog tick, Dermacentor variabilis. AB - Hemolymph from partially fed virgin Dermacentor variabilis females was collected following Borrelia burgdorferi challenge and assayed for antimicrobial activity against Bacillus subtilis and B. burgdorferi. A small inducible cationic peptide was identified by SDS-PAGE in the hemolymph of these ticks as early as 1h post challenge. Following purification by a three-step procedure involving sequential SepPak elution, reversed phase high performance liquid chromatography (RP-HPLC) and gel electrophoresis, the yield of the active peptide was approximately 0.1% of the total protein in the hemolymph plasma. The molecular weight, 4.2kDa, was determined by MALDI-TOF mass spectrometry. N-terminal sequencing by the Edman degradation method gave a sequence for the first 30 amino acids as: G-F-G-C-P-L-N Q-G-A-C-H-N-H-C-R-S-I-(R)-(R)-(R)-G-G-Y-C-S-Q-I-I-K. A computer search of databases showed that the peptide had 83% similarity to a defensin found in a scorpion. This is the first report of a defensin from a tick. The peptide was stable at least up to 70 degrees C. Although the tick defensin alone was not immediately effective against B. burgdorferi, tick defensin plus lysozyme killed more than 65% of cultured B. burgdorferi within 1h. PMID- 11439246 TI - Heterologous expression and characterization of alternatively spliced glutathione S-transferases from a single Anopheles gene. AB - Three cDNA sequences of glutathione S-transferase (GST), adgst1-2, adgst1-3 and adgst1-4, which are alternatively spliced products of the adgst1AS1 gene, were obtained from fourth instar larvae of Anopheles dirus mosquito by reverse transcriptase PCR reactions. The nucleotide sequences of these three cDNAs share >67% identity and the translated amino acid sequences share 61-64% identity. A comparison of the An. dirus to the An. gambiae enzymes shows that adGST1-2 versus agGST1-4, adGST1-3 versus agGST1-5 and adGST1-4 versus agGST1-3 have 85, 92 and 85% amino acid sequence identity, respectively, which confirms that orthologous isoenzymes occur across anopheline species. These three proteins were expressed at high levels, approximately 15-20 mg from 200 ml of E. coli culture. The recombinant enzymes were purified by affinity chromatography on an S hexylglutathione agarose column. The subunit sizes of adGST1-2, adGST1-3 and adGST1-4 are 24.3, 23.9 and 25.1 kDa. The recombinant enzymes have high activities with 1-chloro-2,4-dinitrobenzene (CDNB), detectable activity with 1,2 dichloro-4-nitrobenzene but markedly low activity with ethacrynic acid and p nitrophenethyl bromide. adGST1-3 was shown to be the most active enzyme from the kinetic studies. Permethrin inhibition of CDNB activity, at varying concentrations of CDNB, was significantly different, being uncompetitive for adGST1-2, noncompetitive for adGST1-3 and competitive for adGST1-4. In contrast, permethrin inhibition with varying glutathione concentrations was noncompetitive for all three GSTs. Despite the enzymes being splicing products of the same gene and sharing identical sequence in the N-terminal 45 amino acids, these GSTs show distinct substrate specificities, kinetic properties and inhibition properties modulated by the differences in the C-terminus. PMID- 11439247 TI - "Soft"-cuticle protein secondary structure as revealed by FT-Raman, ATR FT-IR and CD spectroscopy. AB - The nature of the interaction of insect cuticular proteins and chitin is unknown even though about half of the cuticular proteins sequenced thus far share a consensus region that has been predicted to be the site of chitin binding. We previously predicted the preponderance of a beta-pleated sheet in the consensus region and proposed its responsibility for the formation of helicoidal cuticle (Iconomidou et al., Insect Biochem. Mol. Biol. 29 (1999) 285). In this study, we examined experimentally the secondary structure of intact and guanidine hydrochloride extracted cuticle and the cuticular protein extract. The studied cuticle came from the larval dorsal abdomen of the lepidopteran Hyalophora cecropia, a classical example of "soft" cuticle. Analysis with FT-Raman, ATR FT IR and CD spectroscopy indicates that antiparallel beta-pleated sheet is the predominant molecular conformation of "soft-cuticle" proteins both in situ in the cuticle and following extraction. It seems that this conformation dictates the modes of chitin-protein interaction in cuticle, in agreement with earlier proposals (Atkins, J. Biosci. 8 (1985) 375). PMID- 11439248 TI - A bacteria-induced, intracellular serpin in granular hemocytes of Manduca sexta. AB - Serine proteinase inhibitors from the serpin superfamily have been identified as hemolymph proteins from several groups of arthropods, including horseshoe crabs, crayfish, and insects. In the tobacco hornworm, Manduca sexta, one group of serpins present in plasma is generated by alternate exon splicing from serpin gene-1. We have identified a second serpin gene from this insect, M. sexta serpin 2. A serpin-2 DNA clone was isolated from a fifth instar larval cDNA library. The full-length cDNA is 1.5 kb long and encodes a protein of 381 amino acid residues. Amino acid sequence comparisons with other invertebrate serpins reveal approximately 25-40% identity with serpin-2. An expressed sequence tag from Bombyx mori, which is very similar to M. sexta serpin-2, was identified, and the corresponding full-length cDNA sequence was determined. This silkworm homolog of serpin-2 is 57% identical to M. sexta serpin-2. Recombinant M. sexta serpin-2 was used as an antigen to generate a rabbit polyclonal antiserum. This antiserum recognized a 43 kDa protein present in hemocytes but absent from plasma. Western and Northern blot results revealed that serpin-2 gene expression increased dramatically after larvae were injected with bacteria. In situ hybridization showed that the serpin-2 mRNA is present in granular hemocytes of immune stimulated larvae. Serpin-2 purified from hemocytes obtained 24 h after injection of larvae with bacteria lacked inhibitory activity for all proteinases tested except for human cathepsin G. The intracellular location of serpin-2 suggests a function for serpin-2 different from the plasma serpin-1 proteins. PMID- 11439249 TI - Molecular cloning of three cDNAs encoding aminopeptidases from the midgut of Helicoverpa punctigera, the Australian native budworm. AB - Three cDNAs encoding aminopeptidases HpAPN1, HpAPN2 and HpAPN3, were isolated from a 5th instar larval midgut cDNA library from Helicoverpa punctigera, the Australian native budworm. The sequences recovered contain open reading frames encoding proteins of 1011, 952, and 1013 amino acids, respectively. All three proteins share the consensus zinc binding/gluzincin motif HEXXHX(18)E and the sequence GAMEN common to gluzincin aminopeptidases. Furthermore, signal peptide sequences and C-terminal hydrophobic regions preceded by three small amino acids qualifying for cleavage and GPI anchor attachment are present in all three protein sequences. Northern blotting results indicate differences in the levels of expression and developmental regulation of all three aminopeptidases. HpAPN1, HpAPN2, and HpAPN3 are more closely related to APNs from other lepidopterans than they are to each other. This report of three different aminopeptidases N in Helicoverpa punctigera adds support to a recent suggestion that at least one gene duplication has taken place in ancestral lepidopterans. The full sequences of the aminopeptidases are available at GENBANK with the following accession numbers: HpAPN1: AF217248, HpAPN2: AF217249, HpAPN3: AF217250. PMID- 11439250 TI - Bacillus thuringiensis Cry1Ac and Cry1Fa delta-endotoxin binding to a novel 110 kDa aminopeptidase in Heliothis virescens is not N-acetylgalactosamine mediated. AB - We determined that Bacillus thuringiensis Cry1Ac and Cry1Fa delta-endotoxins recognize the same 110, 120 and 170 kDa aminopeptidase N (APN) molecules in brush border membrane vesicles (BBMV) from Heliothis virescens. The 110 kDa protein, not previously identified as an APN, contained a variant APN consensus sequence identical to that found in Helicoverpa punctigera APN 2. PCR amplification of H. virescens cDNA based on this sequence and a conserved APN motif yielded a 0.9 kb product that has 89% sequence homology with H. punctigera APN 2. Western blots revealed that the 110 kDa molecule was not recognized by soybean agglutinin, indicating the absence of GalNAc. A 125I labeled-Cry1Ac domain III mutant (509QNR(511)-AAA) that has an altered GalNAc binding pocket (Lee et al., Appl. Environ. Microbiol. 65 (1999) 4513) showed abolished binding to the 120 APN, reduced binding to the 170 kDa APN, and enhanced binding to the 110 kDa APN. Periodate treated H. virescens BBMV blots were also probed with 125I labeled Cry1Ac and 509QNR(511)-AAA toxins. Both toxins still recognized the 110 kDa APN and a >210 kDa molecule which may be a cadherin-like protein. Additionally, 125I (509)QNR(511)-AAA recognized periodate treated 170 kDa APN. Results indicate that the 110 kDa APN is distinct from other Cry1 toxin binding APNs and may be the first described Cry1Ac-binding APN that does not contain GalNAc. PMID- 11439251 TI - Identification and molecular characterisation of a peritrophin gene, peritrophin 48, from the myiasis fly Chrysomya bezziana. AB - The peritrophic matrix lines the midgut of most insects and has important roles in digestion, protection of the midgut from mechanical damage and invasion by micro-organisms. Although a few intrinsic peritrophic matrix proteins have been characterised, no direct homologues of any of these proteins have been found in other insect species, even closely related species, suggesting that the peritrophic matrix proteins show considerable sequence divergence. We now report the identification of the cDNA and genomic DNA sequences of a Chrysomya bezziana homologue of the Lucilia cuprina intrinsic peritrophic matrix protein, peritrophin-48. The gene for C. bezziana peritrophin-48 spans 1315 bp and consists of three exons (65, 560 and 690 bp, respectively) separated by introns of 566 and 72 bp. The transcriptional start site, identified by a consensus of cDNA clones and primer extension analysis, is probably located 58 bp upstream from the start codon. However, there may be multiple start sites for transcription. Two potential TATA boxes and a consensus arthropod transcription initiator are located within 134 bp of sequence upstream of the putative transcriptional start site suggesting that this region contains the gene promoter. Immuno-fluorescence localization demonstrated that C. bezziana peritrophin-48 was localised to the larval peritrophic matrix. Protein fold recognition analysis indicated structural similarities between peritrophin-48 and wheatgerm lectin. As wheatgerm lectin binds chitin, this result suggested that C. bezziana peritrophin-48 may also bind chitin, a constituent of the peritrophic matrix. Chitin binding studies with a recombinant peritrophin-48 protein confirmed that it binds chitin. A Drosophila melanogaster homologue of peritrophin-48 encoded in an EST and a genomic sequence was also identified. The pairwise percentage identities of the deduced amino acid sequences for the peritrophin-48 homologues from the three higher Dipteran species were relatively low, ranging between 32 and 42%. Despite this sequence variability, the predicted structure of these proteins, dictated by five domains, each containing a characteristic distribution of six cysteines, was strictly conserved. It is concluded that considerable sequence variation can be tolerated in this protein because of the constraints imposed on the structure of the protein by an extensive disulphide bonded framework. PMID- 11439253 TI - Environmental risk and (re)action: air quality, health, and civic involvement in an urban industrial neighbourhood. AB - This paper explores the links between (perceived) environmental risk and community (re) action in an urban industrial neighbourhood in Hamilton, Ontario, Canada. In-depth interviews were conducted with residents of an area with a documented history of adverse air quality, in order to determine the relative influence of social capital (networks, norms, and social trust) and place attachment (sense of belonging in a neighbourhood) in deciding to take civic action around this particular environmental issue. The interviews illustrate the complexity of lay understandings of air pollution, and indicate that social capital is a primary contributor to the decision to take certain kinds of action, while attachment to place plays a lesser role. PMID- 11439254 TI - Making the links between community structure and individual well-being: community quality of life in Riverdale, Toronto, Canada. AB - An inquiry into community quality of life was carried out within a framework that recognizes the complex relationship between community structures and individual well-being. Through use of focus groups and key informant interviews, community members, service providers, and elected representatives in a Toronto community considered aspects of their community that affected quality of life. Community members identified strengths of access to amenities, caring and concerned people, community agencies, low-cost housing, and public transportation. Service providers and elected representatives recognized diversity, community agencies and resources, and presence of culturally relevant food stores and services as strengths. At one level, findings were consistent with emerging concepts of social capital. At another level, threats to the community were considered in relation to the hypothesized role neo-liberalism plays in weakening the welfare state. PMID- 11439255 TI - Women's use of contraception in rural India: a village-level study. AB - This paper examines the determinants of contraceptive use among married women in four villages in rural West Bengal, India. It uses primary quantitative data obtained from a survey of 600 women and qualitative data derived from ethnographic methods. Bi- and multi-variate analyses demonstrate that the factors that most influence a woman's use of contraception include her age, the number of living sons she has, and her religious affiliation. The study also shows that the availability and quality of permanent village-based government health care affects the use of modern contraception. The use of temporary family planning methods is negligible in the area. PMID- 11439256 TI - 'Gentle caring experience'. Seeking alternative health care in Canada. AB - The starting point for this paper is a review of the literature, which seeks to explain the use of alternative medicines, therapies and practices in developed countries. Using the Statistics Canada 1996-97 National Population Health Survey- Health File, we then examine the profile of alternative service users. Our analysis shows that use of alternative health care is still limited to a relatively small segment of Canadians whose profile is similar to those in other developed countries. Women are more likely than men to use alternative medicines, therapies and practices, as are those who have higher incomes and are better educated. To move what has been an essentially empirical discussion forward, we explore critiques of conventional medical practice and propose that the analysis of alternative health care be situated within the geographies of consumption. PMID- 11439257 TI - A brief observational measure for urban neighborhoods. AB - There is growing recognition that neighborhood context contributes to the health and well-being of residents over and above individual characteristics and health behaviors. However, few published reports exist of methods for documenting neighborhood characteristics which are easily administered with minimal outlay of resources. In this paper, we present the development of a brief observational method for urban neighborhoods relevant to the health and well-being of families and children. Data from a socieconomically diverse group of urban neighborhoods are used to create theoretically grounded measures of neighborhood context, and the utility of these measures for discriminating between and within urban neighborhoods is demonstrated. PMID- 11439258 TI - Making space for the 'intellectual' in geographies of disability. AB - There is a growing body of work on geographies of deinstitutionalisation and its consequences, but the weight of this scholarship has focussed on people with mental health problems and physically disabled people. With only few exceptions, the 'post asylum geographies' of intellectually disabled people remain neglected by geographers. We advocate a redressing of this imbalance. First, we assess reasons for the relative absence of 'intellectual' considerations in geographies of disability. We then consider ways in which the full spectrum of those with mental differences might be included in geographical research. We conclude that Wolpert's call 25 years ago to 'open closed spaces' is just as applicable to our efforts to conceptually link health, place, and disability, as it is to the material welfare of vulnerable groups in society. PMID- 11439259 TI - Is there a role for GIS in the 'new NHS'? AB - In this paper we draw on the existing literature to highlight the gap between academic health-based applications of Geographical Information Systems (GIS) and their everyday use within the UK National Health Service (NHS). We provide examples of the operational benefits accruing from using GIS in a range of health care applications whilst acknowledging the limited use of such technologies in strategic health tasks. The implications of recent policy changes for future projected use of such systems are discussed before presenting a research agenda for realising GIS potential within the NHS. The paper concludes by re-emphasising the importance of using GIS in strategic health planning contexts in the light of both recent health-care restructuring and new technological developments in the health service. PMID- 11439260 TI - The evolution of congestive heart failure care. PMID- 11439261 TI - Fighting postmenopausal heart disease. PMID- 11439262 TI - Good news on vitamin C... in food. PMID- 11439263 TI - Web-enabled weight loss. PMID- 11439264 TI - Ask the doctor. I've read that defibrillators are being placed on airplanes and in other public places. They cost just a few thousand dollars, so I was thinking about buying one for my home. Is that a good idea? PMID- 11439265 TI - What to look for in a sunscreen. PMID- 11439266 TI - Reader mail. Colonoscopy revisted. PMID- 11439267 TI - Stroke. Preventing a "brain attack". PMID- 11439268 TI - Oral health. Bad breath. PMID- 11439269 TI - Cancer. Why obesity may cause cancer. PMID- 11439270 TI - Recreation. Summertime and the living is risky. PMID- 11439271 TI - By the way, doctor... My father has Parkinson's disease, and it seems to be getting worse. He's having a harder and harder time getting around. His memory has slipped badly. I've read about fetal-cell therapy for Parkinson's, but then heard on the news that it doesn't work--and might even make some people worse. What do you think? PMID- 11439272 TI - Autism -- Part I. PMID- 11439273 TI - Premenstrual mood disturbance. PMID- 11439274 TI - Alzheimer's disease and the sense of smell. PMID- 11439275 TI - Religion and mortality: new data. PMID- 11439276 TI - Is there a "male menopause" - and will hormones help? PMID- 11439277 TI - Chocolate. PMID- 11439278 TI - Medical memo. Smoking, drinking, and thinking. PMID- 11439279 TI - On call. I am 58-year-old diabetic. I've had my PSA checked every year since I turned 50, but I'm now on kidney dialysis. Will dialysis change my PSA results? PMID- 11439280 TI - Glycemic load, diet, and health. PMID- 11439281 TI - Physical activity, Part I. Start with a walk. PMID- 11439282 TI - Ipriflavone not effective for osteoporosis. PMID- 11439283 TI - Hormone replacement. HRT forum: cancer risk. PMID- 11439284 TI - By the way, doctor. Which drugs cause problems in the sun? PMID- 11439285 TI - By the way, doctor. What is Bell's palsy? PMID- 11439286 TI - Synergistic effect of polymorphisms of uncoupling protein 1 and beta3-adrenergic receptor genes on autonomic nervous system activity. AB - OBJECTIVE: To investigate the association of the promoter region -3826 A to G polymorphism of the uncoupling protein 1 (UCP1) gene with autonomic nervous system (ANS) activity and the interaction of the polymorphism with the Trp64Arg polymorphism of the beta3 adrenergic receptor (beta3AR). SUBJECTS: Three-hundred and forty-nine young (mean age 20.4+/-2.1 y old), healthy Japanese males. MEASUREMENTS: DNA was extracted from whole blood and genotyped by polymerase chain reaction restriction fragment length polymorphism. Plasma glucose, plasma insulin and body mass index (BMI) were measured. Frequency of family history of diabetes or obesity was determined by interview. Subjects randomly chosen from each genotype were examined for ANS activity during supine rest and standing by electrocardiogram power spectral analysis of heart rate variability. RESULTS: UCP1 or beta3AR polymorphism was not associated with BMI, plasma glucose, plasma insulin and frequency of family history of diabetes or obesity. The inhibitory effect of UCP1 polymorphism on ANS activity was observed only with occurrence of the variant of beta3AR. The very low frequency component associated with thermoregulation in the sympathetic nervous system of homozygotes of UCP1 (GG) at supine rest was significantly lower than normal (AA, 203.2+/-50.3 vs 462.2+/-83.6 ms(2); mean+/-s.e., P=0.021). A higher response to postural change to standing was also observed in both sympathetic and parasympathetic nervous activities of AA than of GG. CONCLUSION: While UCP1 polymorphism alone does not affect ANS activity, it has a synergistic effect with beta3AR polymorphism in decreasing sympathetic nervous system activity. PMID- 11439287 TI - Systematic screening for mutations in the human necdin gene (NDN): identification of two naturally occurring polymorphisms and association analysis in body weight regulation. AB - BACKGROUND: NDN, which codes for the human necdin protein, is a candidate gene for Prader-Willi syndrome (PWS). One feature of this neurogenetic disorder is hyperphagia resulting in extreme obesity observed later in development. OBJECTIVE AND DESIGN: In this study we have used single-strand conformation polymorphism (SSCP) analysis to identify sequence variants at the human necdin gene. Furthermore we tested whether these variants were associated with obesity in extremely obese German children and adolescents. RESULTS: Two gene variants could be identified: a g.1352T-->C polymorphism in the putative promotor region and a silent g.2311C-->T polymorphism in the coding region. Genotype and allele frequency distribution of both of the polymorphisms were not significantly different between lower and higher body mass index (BMI) subjects. CONCLUSIONS: Hence, it is unlikely that these polymorphisms play a major role in the emergence of juvenile onset human obesity. PMID- 11439288 TI - A new gene related to human obesity identified by suppression subtractive hybridization. AB - OBJECTIVE: The aim of the research was to identify genes specially expressed in the obese state and potentially involved in the pathogenesis of obesity. DESIGN AND SUBJECTS: We used the technique of suppression subtractive hybridization (SSH), which combines subtractive hybridization with PCR, to generate a population of PCR fragments enriched for transcripts of high or low abundance from differentially expressed genes. PolyA+ mRNA was isolated from subcutaneous abdominal adipose tissue of five massively obese (>35 kg/m(2)) and five normal weight (<25 kg/m(2)) women. cDNA generated from RNA pooled from the obese subjects was contrasted by SSH with an excess of pooled cDNA from the normal weight women. RESULTS: Seventy-nine clones were obtained among which one showed by RT-PCR a higher expression in obese than in normal-weight subjects. This gene was shown to be predominantly expressed in adipose tissue in contrast to brain, liver, kidney, heart and skeletal muscle, and was called "Adipogene". No expression was detected in lung, pancreas and placenta. The cDNA was 1.5 kb long with an open reading frame of 1004 nucleotides encoding a protein of 334 amino acids (37 kDa). No significant sequence similarity was found in databanks, except for weak amino acid homologies with prokaryotic AraC/XylS transcriptional regulator family. Adipogene is encoded on chromosome 8, less than 1 centiMorgan (cM) from the beta3 adrenergic receptor (ADRB3) locus. Weak linkages were observed with body mass index (BMI) and three microsatellite markers located within 10 cM of Adipogene, whereas no linkage was observed with Trp64Arg ADRB3 polymorphism using the Quebec Family Study database. CONCLUSION: Using the SSH technique, we have identified a new gene, called Adipogene, which is overexpressed in the adipose tissue of the obese individuals and could be involved in obesity. PMID- 11439289 TI - G-protein beta3 subunit gene splice variant in obesity and overweight. AB - OBJECTIVE: To determine whether the C825T polymorphism of the G-protein beta3 subunit gene (GNB3) is associated with overweight and obesity. This polymorphism leads to a splice variant (Gbeta3-s) with higher activity and very strong association with essential hypertension. DESIGN: A cross-sectional case-control study. SUBJECTS: The sets of affected and control British/European Caucasian subjects used were: (i) an obesity clinic group most of whom had "morbid obesity" (mean body mass index (BMI) for group=43+/-8 kg/m(2)) and non-obese controls (BMI< or =30); (ii) a group of overweight/obese healthy normotensive community volunteers (BMI>25; mean 29+/-5) and controls (BMI< or =25; mean 23+/-1); (iii) a group of overweight/obese hypertensive patients (BMI>25; mean 30+/-4) and lean hypertensive controls (BMI< or =25; mean=23+/-2). MEASUREMENTS: BMI, blood pressure, serum lipids, alleles of GNB3 polymorphism. RESULTS: Compared with control, frequency of the T allele in obese subjects was higher by 12% in (i), 17% in (ii) and 28% in (iii), but the differences were not statistically significant. Slight tracking of the T allele with elevation in BMI was, however, observed, in the obesity clinic group (P=0.018). CONCLUSION: The C825T splice variant of GNB3 makes little if any contribution to obesity in the groups we tested. PMID- 11439290 TI - The effect of physiological levels of glucagon-like peptide-1 on appetite, gastric emptying, energy and substrate metabolism in obesity. AB - OBJECTIVE: Peripheral infusions of glucagon-like peptide-1 (GLP-1) in humans have been shown to inhibit gastrointestinal motility and decrease hunger and energy intake. However, these investigations used supraphysiological doses. The objective of this study was to investigate the effects of a GLP-1 infusion in a physiological dose on appetite sensations, energy intake, gastric emptying, energy and substrate metabolism. METHODS: Eighteen obese men participated in the placebo-controlled, randomized, single-blinded, cross-over study with infusion of GLP-1 or saline. Resting metabolic rate (RMR) and substrate oxidations were measured by ventilated hood before and after an energy-fixed breakfast. Gastric emptying was measured using paracetamol as a marker. Visual analogue scales were used to assess appetite sensations, thirst and comfort throughout the experiment and palatability of the test meals. Blood was sampled for analysis of hormones (GLP-1, GLP-2, glucose-dependent insulinotropic polypeptide (GIP), insulin, glucagon), and substrates (glucose, lactate, non-esterified fatty acids (NEFA), triacylglycerol (TAG)). Ad libitum energy intake at lunch was registered. RESULTS: Following the breakfast, GLP-1 infusion suppressed ratings of hunger and prospective food consumption (P<0.05), whereas all other subjective ratings and ad libitum energy intake were unaffected. RMR, carbohydrate oxidation and gastric emptying rate were lower during the GLP-1 infusion compared with the saline infusion (P<0.001, P<0.05, P<0.0001, respectively). All plasma hormone and substrate profiles, except NEFA, were significantly reduced by GLP-1 (P<0.0001). CONCLUSION: It is concluded that GLP-1 in physiological concentrations powerfully reduces the rate of entry of nutrients into the circulation by a reduction of gastric emptying rate in obese subjects. The effect of GLP-1 on appetite and food intake may be beneficial in weight reduction. PMID- 11439291 TI - A disparity between conventional lipid and insulin resistance markers at body mass index levels greater than 34 kg/m(2). AB - OBJECTIVE: The aim of this study was to examine changes in lipid profile and markers of insulin resistance with increasing body mass index (BMI) in the range 34-77 kg/m(2). In addition we compare the lipid profiles of severely obese patients with those of the Australian community. SUBJECTS AND METHODS: A total of 572 patients (85% F, 15% M) were assessed prior to gastric restrictive surgery. Conventional lipid profiles and markers of insulin resistance were measured. Lipids were compared with the Australian National Heart Foundation 1989 study (control group). RESULT: There was no difference in mean total cholesterol levels between the obese group (5.52 mmol/l) and the control group (5.47 mmol/l). The mean total cholesterol levels in the obese group fell with increasing BMI (r= 0.13, P<0.01). Obese subjects had elevated fasting triglyceride levels 1.96 mmol/l (control group, 1.12 mmol/l, P<0.001), but levels did not change with increasing BMI (r=0.0, NS). HDL-C levels were lower, 1.21 mmol/l (control group 1.44 mmol/l, P<0.001), and decreased with increasing BMI (r=-0.20, P<0.01). LDL-C levels were lower in obese men (3.65 mmol/l vs control group 4.17 mmol/l, P<0.01) but not women and levels fell with increasing BMI (r=-0.15, P<0.05). For the obese group, markers of insulin resistance (fasting plasma glucose, HbA1c, fasting plasma insulin and C-peptide) all rose significantly with increasing BMI. CONCLUSION: Raised total cholesterol is not a co-morbidity of severe obesity. There is a disparity between the conventional lipid measures and insulin resistance measures of the metabolic syndrome with increasing BMI. Conventional lipid measures may be poor indicators of dyslipidaemic risk in the severely obese. PMID- 11439292 TI - Lack of insulin inhibition on insulin secretion in non-diabetic morbidly obese patients. AB - OBJECTIVE: Insulin inhibition of insulin secretion has been described in normal lean subjects. In this study, we examined whether this phenomenon also occurs in the morbidly obese who often have severe peripheral insulin resistance. SUBJECTS: Twelve obese patients, normotolerant to glucose (8 F/4 M, body mass index (BMI)=54.8+/-2.5 kg/m(2), 39 y) and 16 lean control subjects (10 F/6 M, BMI=22.0+/-0.5 kg/m(2), 31 y). DESIGN AND MEASUREMENTS: An experimental study using various parameters, including an euglycemic hyperinsulinemic clamp (280 pmol/min/m(2) of body surface), an oral glucose tolerance test (OGTT), electrical bioimpedance and indirect calorimetry. RESULTS: The obese subjects were insulin resistant (M=19.8+/-1.6 vs 48.7+/-2.6 micromol/min kg FFM, P<0.0001) and hyperinsulinemic in the fasted state and after glucose ingestion. Fasting plasma C-peptide levels (obese 1425+/-131 pmol/l vs lean 550+/-63 pmol/l; P<0.0001) decreased less during the clamp in the obese groups (-16.9+/-6.9% vs -43.0+/-5.6% relative to fasting values; P=0.007). In the lean group, the C-peptide decrease during the clamp (percentage variation) was related to insulin sensitivity, M/FFM (r=0.56, P=0.03), even after adjustment for the clamp glucose variation. CONCLUSION: We conclude that, in lean subjects, insulin inhibits its own secretion, and this may be related to insulin sensibility. This response is blunted in morbidly obese patients and may have a role in the pathogenesis of fasting hyperinsulinemia in these patients. PMID- 11439293 TI - Plasma leptin is independently associated with the intima-media thickness of the common carotid artery. AB - OBJECTIVE: To investigate whether intima-media thickness (IMT) of the common carotid artery (CCA), an early marker of asymptomatic atherosclerosis, is significantly and independently associated with plasma concentrations of leptin, an adipose tissue hormone that has recently been proposed as a cardiovascular risk factor in obese patients. DESIGN: Cross-sectional sample of normal-weight and obese men and women. SUBJECTS: One-hundred and twenty healthy subjects (52 men and 68 women), aged 18-45 y and with a wide range of BMI, were recruited for the study. MEASUREMENTS: Fasting plasma leptin concentrations and the IMT of the CCA were measured in all subjects. Leptin concentrations were measured by radioimmunoassay and the IMT of the CCA was quantified by high resolution B-mode ultrasound imaging. Central fat (measured by waist circumference), smoking habits, blood pressure, insulin sensitivity (measured by the insulin tolerance test), and fasting plasma glucose, insulin and lipid pattern (cholesterol, HDL cholesterol, triglycerides, LDL-cholesterol) were also measured. RESULTS: IMT of the CCA was positively correlated with log leptin concentrations (P<0.005 in men and P<0.001 in women), body mass index (P<0.001 in men and women), waist circumference (P<0.001 in men and women), age (P<0.001 in men and P<0.05 in women), and negatively associated with insulin sensitivity in both sexes (P<0.05). IMT was also directly correlated with cholesterol (P<0.05), LDL cholesterol (P<0.01) and systolic blood pressure in men (P<0.05), and with diastolic blood pressure levels in women (P<0.05). When a multiple linear regression model was used without body mass index (BMI), the correlation between leptin and IMT was maintained in both men (P<0.01) and women (P<0.005), independent of age, insulin sensitivity, smoking habits, systolic blood pressure, fasting glucose, triglycerides, cholesterol, LDL-cholesterol and HDL-cholesterol. By contrast, BMI-adjusted leptin concentrations were not significantly associated with IMT (Pc (partial correlation): 0.41 in men and 0.15 in women). Moreover, when BMI was entered into a multiple linear regression model without leptin, the correlation between BMI and IMT was maintained in both men (P<0.005) and women (P<0.01), independent of the same parameters. CONCLUSION: Plasma leptin concentrations are independently associated with the IMT of the CCA, suggesting that the increase of adipose tissue mass (or leptin per se) may have an unfavourable influence on the development of atherosclerosis. However, the association between IMT and leptin seems to be dependent and/or confounded by the relationship between IMT and obesity. PMID- 11439294 TI - Increased expression of eNOS protein in omental versus subcutaneous adipose tissue in obese human subjects. AB - OBJECTIVE: To investigate the expression of eNOS and iNOS mRNA and protein in adipose tissue from subcutaneous (s.c.) and omental adipose tissue of obese subjects. DESIGN: Subcutaneous and omental adipose tissue was obtained from subjects undergoing weight reduction surgery. Messenger RNA and protein levels were measured in tissue extracts and related to basal lipolysis, which was measured in isolated adipocytes from the same subjects. SUBJECTS: Eight overweight but otherwise healthy male subjects (age 43.4+/-10.3 y, BMI 39+/-3.5 kg/m(2), mean+/-s.e.m.). MEASUREMENTS: For mRNA detection a competitive reverse transcription polymerase chain reaction method was used while protein was detected by Western blot. Glycerol release was determined in isolated adipocytes using a standard luminometric assay. RESULTS: Tissue mRNA levels for eNOS in s.c. tissue were 6098+/-1969 amol/mg RNA and in omental tissue 6987+/-2914 amol/mg RNA (mean+/-s.e.m., P=0.75). iNOS mRNA levels were substantially lower; in s.c. tissue 227+/-127 amol/mg RNA and in omental tissue 245+/-162 amol/mg RNA (P=0.8). In Western blot, eNOS protein levels in s.c. and omental tissue were 1.88+/-2.0 and 7.47+/-4.11 (OD/mm(2) 100 microg total protein, P=0.0063), respectively. iNOS protein was expressed at significantly lower levels and barely detectable in both s.c. and omental tissue. Basal rate of lipolysis was two times higher in s.c. compared to omental fat cells (P=0.028). CONCLUSIONS: eNOS protein is markedly increased in omental compared to s.c. adipose tissue in human obese subjects, probably due to post-transcriptional mechanisms. Since basal lipolysis is much lower in omental vs s.c. adipose tissue it is possible that regionally increased NO production, primarily by eNOS, may be involved in the site difference of basal lipolysis in obese subjects. PMID- 11439295 TI - Long-term leptin treatment of ob/ob mice improves glucose-induced insulin secretion. AB - OBJECTIVE: Previous studies have demonstrated that leptin inhibits glucose stimulated insulin secretion from isolated islets, although a lack of leptin effect on insulin secretion has also been reported. The effect of long term in vivo leptin treatment of insulin secretion has, however, not been established. Therefore, in the present study, we have evaluated the effect of long term in vivo treatment of leptin on glucose-induced insulin secretion in ob/ob mice. METHODS: After 7 days' treatment of leptin (100 microg daily s.c.), insulin release was measured in isolated islets by batch incubation followed by radioimmunoassay. Glucose utilization and oxidation were measured by measuring the formation of (3)H(2)O and (14)CO(2) from [5-(3)H] and [U-(14)C] glucose, respectively. Glucose-6-phosphatase activity was measured by measuring the conversion of (14)C-glucose-6-P to (14)C-glucose. In addition, immunohistochemistry of pancreatic specimens was undertaken for study of expression of insulin, GLUT-2 and hormone-sensitive lipase (HSL). RESULTS: Leptin treatment significantly improved insulin secretion both at 5.5 mM (by 15%; P<0.05) and 16.7 mM (by 85%; P<0.001) glucose, compared to vehicle-treated controls. Furthermore, whereas leptin treatment did not affect islet insulin or DNA contents, a significant decrease in islet triglyceride content and glucose-6 phosphatase activity was observed. Moreover, the immunocytochemical data revealed an increased immunostaining for insulin, GLUT-2 and hormone-sensitive lipase (HSL) in islets from leptin-treated ob/ob mice. CONCLUSION: The results suggest that long-term leptin treatment of ob/ob mice improves glucose-stimulated insulin secretion in parallel with reduced glucose-6-phosphatase activity, increased HSL and decreased triglyceride levels in islets. These perturbations may explain the improvement of glucose-stimulated insulin secretion induced by leptin. PMID- 11439296 TI - Physical activity and determinants of physical activity in obese and non-obese children. AB - OBJECTIVE: To compare the physical activity (PA) patterns and the hypothesized psychosocial and environmental determinants of PA in an ethnically diverse sample of obese and non-obese middle school children. DESIGN: Cross-sectional study. SUBJECTS: One-hundred and thirty-three non-obese and 54 obese sixth grade children (mean age of 11.4+/-0.6). Obesity status determined using the age-, race and gender-specific 95th percentile for BMI from NHANES-1. MEASUREMENTS: Objective measurements were collected of PA over a 7-day period using the CSA 7164 accelerometer: total daily counts; daily moderate (3-5.9 METs) physical activity (MPA); daily vigorous physical activity (> or =6 METs; VPA); and weekly number of 5, 10 and 20 min bouts of moderate-to-vigorous physical activity (> or =3 METs, MVPA). Self-report measures were collected of PA self-efficacy; social influences regarding PA, beliefs about PA outcomes; perceived PA levels of parents and peers, access to sporting and/or fitness equipment at home, involvement in community-based PA organizations; participation in community sports teams; and hours spent watching television or playing video games. RESULTS: Compared to their non-obese counterparts, obese children exhibited significantly lower daily accumulations of total counts, MPA and VPA as well as significantly fewer 5, 10 and 20 min bouts of MVPA. Obese children reported significantly lower levels of PA self-efficacy, were involved in significantly fewer community organizations promoting PA and were significantly less likely to report their father or male guardian as physically active. CONCLUSIONS: The results are consistent with the hypothesis that physical inactivity is an important contributing factor in the maintenance of childhood obesity. Interventions to promote PA in obese children should endeavor to boost self efficacy perceptions regarding exercise, increase awareness of, and access to, community PA outlets, and increase parental modeling of PA. PMID- 11439297 TI - Parameters of childhood obesity and their relationship to cardiovascular risk factors in healthy prepubescent children. AB - OBJECTIVE: To investigate which of the currently applied parameters to assess childhood overweight best predict cardiovascular risk factors. DESIGN: Cross sectional study comparing five different methods to define overweight with respect to their power to predict cardiovascular risk factors. SUBJECTS: A total of 838 healthy children from the Prevention-Education-Program (Nuremberg, Germany; age 4-9 y, 405 boys, 433 girls). MEASUREMENTS: Obesity parameters-body mass index (BMI), ponderal index (PI), the sum of triceps and subscapular skinfold thickness (SFT), percentage body fat (%BF) using SFT and two different regression formulas (Slaughter, %BF-SL; Dezenberg, %BF-DZ). Overweight defined by the 90th age- and sex-specific percentile of each obesity parameter. Comparison of LDL- and HDL-cholesterol, apolipoprotein-B (apo-B), triglycerides (TG), fibrinogen and blood pressure values (SBP/DBP) between normal-weight and overweight children. RESULTS: When overweight is defined by BMI or PI, all cardiovascular risk factors are significantly (P<0.01) different between overweight and normal-weight children (BMI: TG+20.5%, HDL-chol.-8.6%, LDL chol.+9.6%, apo-B+6.8%, SBP+7.4%, DBP+8.6%, fibrinogen+13.2%; PI: TG+24.3%, HDL chol.-6.1%, LDL-chol.+9.0%, apo-B+7.4%, SBP+5.9%, DBP+6.7%, fibrinogen+13.9%), while SFT, %BF-SL and %BF-DZ did not predict all cardiovascular risk factors. A sex-specific analysis showed that in girls BMI and PI both predict cardiovascular risk factors, while in boys this is only valid for BMI. CONCLUSION: In prepubescent children, height-to-weight indices such as BMI or PI better predict cardiovascular risk factors than obesity parameters using skinfold measurement. The BMI may be superior to the PI as the association between BMI and cardiovascular risk factors is less affected by gender. PMID- 11439298 TI - A genetic analysis of weight and overweight in 4-year-old twin pairs. AB - OBJECTIVE: Although many twin and adoption studies document genetic influence on individual differences in weight, much less is known about genetic influences on overweight, about the genetic links between weight and overweight, or about the origins of weight and overweight in childhood, an age that might provide a good target for prevention of obesity. We tested the hypothesis that, in early childhood, overweight is as heritable as weight and that weight and overweight are linked genetically. DESIGN: Model-fitting analyses were used to compare monozygotic (MZ) and dizygotic (DZ) twins (same-sex and opposite-sex) for weight and overweight. SUBJECTS: The sample included 3636 4-y-old twins born in the UK in 1994. MEASUREMENTS: Heights and weights reported by parents were used to assess weight corrected for height, which yields results similar to body mass index (BMI) but corrects more completely for genetic effects on height. RESULTS: At 4 y of age, genetic factors contributed substantially both to individual differences in weight throughout the distribution and to the mean weight difference between overweight children and the rest of the population. Unlike results later in life, weight and overweight in 4-y-olds also suggest substantial shared family environmental influence. Results are similar for boys and girls. CONCLUSIONS: Overweight is the quantitative extreme of genetic and environmental factors responsible for normal variation in weight in childhood. Genes associated with overweight are likely to be associated with variation in weight throughout the distribution, as assumed by quantitative trait locus (QTL) theory. These findings linking weight and overweight in childhood have far-reaching implications for molecular genetic attempts to identify specific genes responsible for genetic influence, for investigating pathways between genes and behaviour, and for intervention and prevention. PMID- 11439299 TI - Does obesity influence foot structure and plantar pressure patterns in prepubescent children? AB - OBJECTIVE: This study examined the effects of obesity on plantar pressure distributions in prepubescent children. DESIGN: Field-based, experimental data on BMI (body mass index), foot structure and plantar pressures were collected for 13 consenting obese children and 13 non-obese controls. SUBJECTS: Thirteen obese (age 8.1+/-1.2 y; BMI 25.5+/-2.9 kg/m(2)) and 13 non-obese (age 8.4+/-0.9 y; BMI 16.9+/-1.2 kg/m(2)) prepubescent children, matched to the obese children for gender, age and height. MEASUREMENTS: Height and weight were measured to calculate BMI. Static weight-bearing footprints for the right and left foot of each subject were recorded using a pedograph to calculate the footprint angle and the Chippaux-Smirak index as representative measures of the surface area of the foot in contact with the ground. Right and left foot plantar pressures were then obtained using a mini-emed(R) pressure platform to calculate the force and pressure experienced under each child's foot during static and dynamic loaded and unloaded conditions. RESULTS: Obese subjects displayed significantly lower footprint angle (t=4.107; P=<0.001) values and higher Chippaux-Smirak index values (t=-6.176; P=<0.001) compared to their non-obese counterparts. These structural foot changes were associated with differences in plantar pressures between the two subject groups. That is, although rearfoot dynamic forces generated by the obese subjects were significantly higher than those generated by the non-obese subjects, these forces were experienced over significantly higher mean peak areas of contact with the mini-emed(R) system. Therefore, rearfoot pressures experienced by the two subject groups did not differ. However, the mean peak dynamic forefoot pressures generated by the obese subjects (39.3+/-15.7 N.cm(-2); q=3.969) were significantly higher than those generated by the non obese subjects (32.3+/-9.2 N.cm(-2)). CONCLUSIONS: It is postulated that foot discomfort-associated structural changes and increased forefoot plantar pressures in the obese foot may hinder obese children from participating in physical activity and therefore warrants immediate further investigation. PMID- 11439300 TI - Does the pattern of postpartum weight change differ according to pregravid body size? AB - OBJECTIVES: To examine differences in the pattern of weight changes during and after pregnancy among four pregravid body mass index (BMI) groups. STUDY DESIGN: Prospective cohort study of women who had two consecutive births at the University of California, San Francisco (UCSF) between 1980 and 1990. MEASUREMENTS: Maternal body weights were available before conception and delivery, and at 6 weeks postpartum for the first (index) pregnancy, and before conception for the second study pregnancy. Height and two pregravid weights were self-reported. Weights at delivery and 6 weeks postpartum were measured. Net delivery weight was defined as delivery weight minus infant birth weight. Three non-overlapping sequential weight changes were constructed: (1) net gestational gain (net delivery weight minus pregravid weight at the index pregnancy); (2) early net postpartum weight change (6-week postpartum weight minus net delivery weight); and (3) late postpartum weight change (pregravid weight at the second pregnancy minus 6-week postpartum weight). SUBJECTS: A total of 985 healthy women (age 18-41 y) from four race/ethnicity groups (Asian, Hispanic, black and white) who had a singleton, full-term, live birth for the index pregnancy followed by a second consecutive birth. RESULTS: Four race/ethnicity groups were combined (no interaction) to contrast average weight changes among pregravid BMI groups. Means adjusted for eight covariates (parity, race/ethnicity, education, mode of delivery, smoking, hypertension of pregnancy, age, height) and time intervals were not altered appreciably. Early net postpartum weight losses were similar for all pregravid BMI groups. Late (median of 2 y) postpartum weight losses were 4 kg higher in the low and average BMI groups compared with the highest BMI group. About half of the net gestational gain was lost by 6 weeks postpartum, and the percentage that was lost decreased over time. CONCLUSIONS: This study suggests that early postpartum weight loss does not vary by maternal pregravid BMI group, but late postpartum weight change does. Serial weight measurements are needed in epidemiologic studies to differentiate retention of gestational gain from weight gain during the late postpartum period. PMID- 11439301 TI - Body mass index in mid-life women: relative influence of menopause, hormone use, and ethnicity. AB - OBJECTIVE: To assess the relative influence of menopausal status and hormone use on body mass index (BMI) among a multiethnic sample of mid-life women. DESIGN: Cross-sectional telephone survey conducted at seven sites where each site targeted an ethnic minority group and Caucasians as part of Study of Women's Health Across the Nation (SWAN). SUBJECTS: A total of 7181 Caucasians, 3949 African-Americans, 1660 Hispanics, 562 Chinese Americans, and 803 Japanese Americans between ages of 40 and 55 y residing in or near Boston, Chicago, Detroit, Los Angeles, Newark, NJ, Oakland, CA, and Pittsburgh, PA. MEASUREMENTS: Self-reported BMI based on weight in kg divided by height in m(2) menopausal status, physical inactivity, postmenopausal hormone use, ethnicity, and age in years. RESULTS: Compared to premenopausal women (covariate adjusted M=27.3), women reporting a surgical menopause (M=28.2) or being in the perimenopausal transition (M=27.7 for early and 27.9 for late perimenopause) had higher BMI. Women reporting a natural menopause (M=27.4) did not have a higher BMI than premenopausal women, after adjusting for chronological age and other covariates. Hormone use was associated with lower BMI (M=26.5 vs 27.3). A comparison of effect sizes showed that menopausal status (F=13.1), followed by chronological age (F=24.0), were the least powerful predictors of BMI, whereas the more powerful predictors were physical activity level (F=1377.1) and ethnicity (F=400.5). CONCLUSIONS: The menopausal transition affects body mass index in mid life, but the effect is small relative to other influences. Interventions to increase physical activity are highly recommended to prevent increases in adiposity common in mid-life. PMID- 11439302 TI - Hormones, weight change and menopause. AB - OBJECTIVE: To determine total body weight change occurring in women at mid-life, specifically with respect to occurrence of menopause and use of estrogen. DESIGN: Retrospective analysis of body weight measurements accumulated in two cohorts of healthy women participating in studies of skeletal metabolism. SUBJECTS: Cohort 1: 191 healthy nuns enrolled in a prospective study of osteoporosis risk, aged 35 45 in 1967; cohort 2: 75 women aged 46 or older and still menstruating, enrolled in 1988 in a study of bone cell dynamics across menopause. Roughly one-third of each group received hormone replacement after menopause. MEASUREMENTS: Body weight and height, age, menstrual status and use of estrogen replacement. Cohort 1: 608 measurements at 5 y intervals spanning a period from 17 y before to 22 y after menopause; cohort 2: 1180 measurements at 6-month intervals spanning a period from 5 y prior to 5 y after menopause. RESULTS: In cohort 1 weight rose as a linear function of age (both chronological and menopausal), both before and after cessation of ovarian function, at a rate of approximately 0.43% y(-1). Neither the menopausal transition nor the use of estrogen had an appreciable effect on this rate of gain. In cohort 2 the rate of gain seemed to diminish slightly at menopause. As with cohort 1, hormone replacement (or its absence) had no appreciable effect on weight. CONCLUSIONS: The long-term, total body weight trajectory at mid-life is not influenced appreciably by either cessation of ovarian function or by hormone replacement. PMID- 11439303 TI - Relationship between total and regional bone mineral density and menopausal state, body composition and life style factors in overweight Japanese women. AB - OBJECTIVE: To investigate whether menopausal state, body composition and lifestyle factors influence total and regional bone mineral density in overweight Japanese women. DESIGN: Cross-sectional study of women who were recruited to the weight reduction program held at community-based health promotion center in Tokyo area. SUBJECTS: A total of 178 women with a mean age of 48 y old (20-69 y) with a clear menstrual history and BMI over 24. MEASUREMENTS: Total, regional and lumbar spine bone mineral density (BMD) and body composition were measured using DXA (Lunar). Menstrual history was taken by a questionnaire and walking steps per day and energy intake were measured. Physical fitness was assessed by cardio respiratory fitness and leg extension power. Subjects were divided into pre menopausal and post-menopausal groups. RESULTS: Pre-menopausal group had significantly higher total body BMD as well as regional BMD than post-menopausal group. However, no differences in BMI, percentage fat and fat mass (FM) were seen between the two groups. The multiple regression analysis stepwise method revealed that total and regional BMD correlated with menopausal state and total FM independently. Total and regional BMD did not correlate with total non-fat soft tissue mass (NFSM), energy intake, walking steps or physical fitness levels. Trunk and lower extremities BMD correlated with corresponding regional FM and NFSM, and upper extremities BMD correlated with only corresponding body part NFSM after adjusting menopausal state. CONCLUSION: Total and regional BMD had strong negative correlation with menopausal state rather than total FM in overweight Japanese women. Weight-bearing site BMD correlated with corresponding body part FM and NFSM and non-weight bearing site BMD only correlated with corresponding body part NFSM after adjusting for menopausal state. PMID- 11439304 TI - Tall stature, overweight and the prevalence of low back pain in Chinese middle aged women. AB - OBJECTIVES: First, to estimate the rate and characteristic of low back pain (LBP) among Hong Kong middle-aged women. Second, to explore the relationship between being overweight, having tall stature and LBP prevalence. DESIGN: A case-control study of Hong Kong community-based middle-aged women was conducted. SUBJECTS: The study subjects were either from the University Family Medicine Clinic or from a population-based cross-sectional study on health in middle-aged women conducted by the Department of Community and Family Medicine (based on a randomized sample of telephone numbers from the Hong Kong residential telephone directory in 1991). Among the 928 potential eligible subjects from both sources, a total of 182 cases and 235 controls participated in this study. MEASUREMENTS: Data were collected from personal interviews and anthropometric measurements. Participants were asked about exclusion criteria, the occurrence of LBP and demographics factors. The anthropometric measurements included measures of body height, body weight, waist and hip circumferences. RESULTS: Among the 182 cases who entered this study, 83 women (45.6%) had 1-<14 days of LBP, and 99 women (54.4%) had at least 14 days of LBP in the previous 12 months. Having tall stature and being overweight were found to have no association with LBP prevalence. Conversely, high waist-to-hip ratio (> or =0.8) was noted to be inversely associated with the risk of severe (at least 14 days) LBP (adjusted odds ratio=0.43, 95% confidence interval 0.26 0.70) even when controlling for other risk factors together, but not for the overall (at least one day) LBP. CONCLUSION: This case-control study found no association between excessive weight, tall stature and an increased risk of LBP prevalence in Hong Kong Chinese middle-aged women. Conversely, the results indicate that a high waist to hip ratio was associated with a lower risk of severe LBP. Prospective studies are needed to further evaluate the association between underweight/obesity, bone mass density and LBP. PMID- 11439305 TI - Functional limitations linked to high body mass index, age and current pain in obese women. AB - OBJECTIVE: To describe functional limitations in obese women. DESIGN: Comparisons of functional limitations in obese women and in a normal-weight reference group regarding mobility, balance and housework transport. SETTING: A large room with a corridor and staircase nearby. SUBJECTS: Fifty-seven consecutively selected obese female outpatients, mean age 44 y, body mass index (BMI) 37 kg.m(-2), and 22 voluntary references, mean age 49 y, BMI 22 kg.m(-2). MAIN OUTCOME MEASURES: Pain questionnaire (for background data); a new valid and reliable test protocol measuring activities of daily life (ADL), pain and exertion, heart rate, balance and speed. RESULTS: The patients had problems in performing 13 of 16 actions compared with the references (P-values between 0.05 and 0.001). They moved more slowly and had difficulties and /or pain and exertion in flexibility tasks, balancing, activities at floor level (bending and kneeling), climbing stairs and carrying groceries. BMI (P=0.002), age (P=0.02) and current pain (P=0.02) from the lower body predicted functional limitation. CONCLUSIONS: The functional test reflected functional limitations in obese women, revealing difficulties and perceived pain in ADL. The test protocol can be used to evaluate the effects of training or dieting. For clinical use we recommend considering BMI, age and current pain for use when individualizing advice on training. PMID- 11439306 TI - The measurement of dietary restraint, disinhibition and hunger: an examination of the factor structure of the Three Factor Eating Questionnaire (TFEQ). AB - OBJECTIVE: To conduct separate factor analyses of the Three Factor Eating Questionnaire (TFEQ-R, TFEQ-D and TFEQ-H) scales and provide initial evidence of the construct validity of the obtained solutions. DESIGN: A cross-sectional survey with a 12 month retest of a subsample of subjects. SUBJECTS: A total of 553 undergraduate university women with a mean age of 25.0 y. The retest sample comprised 64 subjects with a mean age at retest of 25.7 y. MEASUREMENTS: In addition to the TFEQ, age, body mass index (BMI), satisfaction with current weight, nutrition knowledge and current exercise level were recorded. RESULTS: Three restraint (strategic dieting behaviour, attitude to self-regulation, avoidance of fattening foods), three disinhibition (habitual susceptibility, emotional susceptibility, situational susceptibility) and two hunger constructs (internal locus for hunger, external locus for hunger) were identified. Initial evidence of the validity of these constructs was provided. CONCLUSION: The explanation of disordered eating behaviour is likely to be refined more by specific constructs, such as those presented, rather than by the more general constructs measured by the original TFEQ-R, TFEQ-D and TFEQ-H scales. Further examination of the factor structures presented is therefore encouraged. PMID- 11439307 TI - Self-efficacy in relation to eating behaviour among obese and non-obese women. AB - OBJECTIVE: To assess the effect of a 3 month behaviour modification weight management programme on self-efficacy and anthropometric variables among obese women seeking treatment at an obesity management clinic and to compare self efficacy among these obese women to non-obese women. DESIGN: Cross sectional. SUBJECTS: A total of 161 non-obese (BMI 22.6+/-2.9 kg/m(2)) and 138 obese (BMI 37.7+/-5.8 kg/m(2)) women of similar age. MEASUREMENTS: Self-efficacy in relation to eating was assessed by the Weight Efficacy Lifestyle (WEL) questionnaire. Demographic information was obtained by interview and questionnaire in the obese and by questionnaire in the non-obese. Anthropometric measurements were obtained by direct measure in the obese and BMI was calculated from self-reported weight and height in the non-obese. RESULTS: At entry to the programme obese women scored significantly less (P<0.0001) than non-obese women on the WEL (99.4+/-34.1 vs 139.0+/-24.9). Women who completed the programme (n=65) demonstrated a decrease in waist circumference of 3.9+/-5.3 cm, a 10.0+/-11.5% loss of excess weight and a significant improvement in total WEL score from 106.0+/-30.3 to 126.5+/-28.4. CONCLUSION: Improvements in some dimensions of self-efficacy among obese women were of sufficient magnitude to attain scores similar to women of a normal weight. The WEL questionnaire may provide an additional measure of success as well as provide positive feedback and encouragement to the client. PMID- 11439308 TI - Associations of physical activity with body weight and fat in men and women. AB - OBJECTIVE: Increasing physical activity is strongly advocated as a key public health strategy for weight gain prevention. We investigated associations of leisure-time physical activity (LTPA) and occupational/domestic physical activity with body mass index (BMI) and a skinfold-derived index of body fat (sum of six skinfolds), among normal-weight and overweight men and women. DESIGN: Analyses of cross-sectional self-report and measured anthropometric data. SUBJECTS: A total of 1302 men and women, aged 18-78 y, who were part of a randomly selected sample and who agreed to participate in a physical health assessment. MEASUREMENTS: Self report measures of physical activity, measured height and weight, and a skinfold derived index of body fatness. RESULTS: Higher levels of LTPA were positively associated with the likelihood of being in the normal BMI and lower body fat range for women, but few or no associations were found for men. No associations were found between measures of occupational/domestic activity and BMI or body fat for men or women. CONCLUSION: By using a skinfold sum as a more direct measure of adiposity, this study extends and confirms the previous research that has shown an association between BMI and LTPA. Our results suggest gender differences in the relationship of leisure-time physical activity with body fatness. These findings, in conjunction with a better understanding of the causes of such differences, will have important public health implications for the development and targeting of weight gain prevention strategies. PMID- 11439309 TI - A nutrition and exercise intervention program for controlling weight in Samoan communities in New Zealand. AB - OBJECTIVE: To promote weight loss in Samoan church communities through an exercise program and nutrition education. METHODS: A quasi-experimental design was used to assess weight change, over 1 y, in cohorts of people aged 20-77 y from three non-randomised Samoan church communities (two intervention, n=365 and one control, n=106) in Auckland, New Zealand. The intervention churches received aerobics sessions and nutrition education about dietary fat. RESULTS: Baseline body mass index for the intervention and control churches was (mean+/-s.e.) 34.8+/-0.4 and 34.3+/-0.9 kg/m(2), respectively. The intervention churches lost an average of 0.4+/-0.3 kg compared to a 1.3+/-0.6 kg weight gain in the control church (P=0.039, adjusted for confounders). The number of people who were vigorously active increased by 10% in the intervention churches compared to a 5% decline in the control church (P=0.007). Nutrition education had little apparent impact on knowledge or behaviour. CONCLUSION: Samoan communities in New Zealand are very obese and have high rates of annual weight gain. A community-based intervention program arrested this weight gain in the short term. PMID- 11439310 TI - Alcohol consumption and blood pressure change: 5-year follow-up study of the association in normotensive workers. AB - This study endeavours to clarify the effect of alcohol consumption on changes in the blood pressure of Japanese men. The subjects, who were followed from 1990 to 1995, were 2189 male normotensive steelworkers aged from 40 to 54 years in 1990. Drinking habits were represented by two variables: average daily baseline alcohol consumption, and change in alcohol consumption during the observation period, both derived from self-report questionnaires and interviews. Nine other items were analysed as covariates: systolic and diastolic blood pressure, age, body mass index (BMI), salt intake, physical activity, and levels of gamma glutamyl transpeptidase (GTP), uric acid and plasma glucose in 1990. The association between drinking habits and blood pressure was evaluated using ANCOVA. Subjects who had initiated antihypertensive therapy by 1995 were excluded from this dataset. The results of analysis disclosed that both daily alcohol consumption and its change were significantly associated with changes in systolic and diastolic blood pressure during the observation period. J-shaped associations of daily alcohol consumption with adjusted changes in systolic and diastolic blood pressure were found. Positive associations were observed between changes in alcohol consumption and adjusted changes in systolic and diastolic blood pressure. In conclusion, daily alcohol consumption was associated non-linearly with changes in blood pressure and with a threshold effect at 18 ml of ethanol per day in these middle-aged Japanese workers. In addition, increasing alcohol consumption was associated with blood pressure elevation, and decreasing consumption was associated with suppression of blood pressure elevation. PMID- 11439311 TI - Long-term diuretic therapy in patients with coronary disease: increased colon cancer-related mortality over a 5-year follow-up. AB - OBJECTIVES: Recent studies have suggested that long-term diuretic therapy may be associated with increased risk of renal cell carcinoma. This carcinoma is not a common malignancy, but it shares risk factors with the considerably more widespread colon cancer (CC). However, there are no data whether or not a relationship between long-term diuretic therapy and CC mortality exists. In this study we tested the hypothesis that long-term diuretic therapy may be associated with increased CC mortality over a 5.6-year follow-up period. SUBJECTS AND METHODS: The study sample comprised 14 166 patients aged 45 to 74 years with a previous myocardial infarction and/or stable anginal syndrome, screened for participation in the bezafibrate infarction prevention (BIP) study. There were 2153 patients receiving diuretics and 12 013 patients receiving no diuretics. RESULTS: During the follow-up 139 (6.5%) new cases of cancer were diagnosed in the diuretic-treated group compared with 622 (5.2%) in the group receiving no diuretics (P = 0.02). Colon cancer mortality was significantly higher in the diuretic-treated patients (0.1 vs 0.5%, P = 0.001), whereas mortality differences for other cancer types were not documented. Multivariate analysis identified diuretics as an independent predictor of increased colon cancer incidence and colon cancer mortality with a hazard ratio (HR) of 2.0 (95% CI 1.2-3.2) for colon cancer incidence and 3.7 (95% CI 1.7-8.3) for mortality. However, the association between diuretic therapy and higher incidence of colon cancer was observed only among non-users of aspirin. A relatively lower colon cancer incidence was observed in the furosemide subgroup, and higher in the small combined amiloride/hydrochlorthiazide subgroup (HR 3.15, 95% CI 1.15-8.65). CONCLUSION: Long-term exposure to diuretic therapy may be associated with an increased colon cancer-related mortality. PMID- 11439312 TI - An observational study of the medical events associated with clinician-initiated changes in treatment for essential hypertension. AB - We report a retrospective longitudinal observational study of co-morbidities and medical events associated with initiations and changes in antihypertensive therapy in 475 hypertensive patients of a large general practice. The median follow-up time was 7.0 years for males and 7.2 years for females. The data showed a low frequency of appropriate lifestyle recommendations (<30%), a gender-bias in lifestyle recommendations against women and that more than half of all patients' blood pressure (BP) was uncontrolled when last seen. Nearly half of all patients had co-morbidities relevant to essential hypertension (EHT) at first treatment for EHT and more than 11% of patients had more than one such co-morbidity. Whilst there was an increase in usage of ACE inhibitors and calcium channel blockers (CCB) as first treatment for EHT, there was also evidence that the existence of relevant co-morbidities rationally accounted for the majority of that increase. There were 5176 medical events relevant to EHT associated with change of drug or dosage treatment of EHT and the study provided evidence that the occurrence of such relevant medical events can rationally account for the majority of changes to EHT treatment. The study suggests that whilst general practitioners may fail to promote lifestyle changes to their patients with EHT, there is evidence that, when examined in sufficient detail, general practitioners' decisions to initiate changes in antihypertensive therapy are in keeping with the evidence base. PMID- 11439313 TI - Antihypertensive efficacy of amlodipine in children with chronic kidney diseases. AB - In adults the calcium antagonist amlodipine given once a day has proved to be an attractive addition to the antihypertensive armamentarium. The present report describes our experience in 43 paediatric outpatients (26 boys and 17 girls, aged between 1.1 and 19, median 9.8 years) with chronic kidney diseases. The patients were given amlodipine for 16 weeks as part of their antihypertensive treatment. Before amlodipine arterial pressure was 150 (142-163)/90 (84-95) mm Hg (median and interquartile range). Six patients withdrew from amlodipine because of oedema, flushing or headache. In the remaining patients amlodipine 7.7 (6.9-9.4) mg/m(2) body surface area once a day significantly decreased arterial pressure by 17 (13-22)/10 (7-13) mm Hg. The efficacy of amlodipine was more pronounced in girls than in boys. No changes in heart rate, body weight and circulating haemoglobin, sodium, potassium and creatinine were noted. In none of the patients circulating potassium, sodium or creatinine changed by more than 0.5 mmol/l, 5 mmol/l respectively 20%. In 11 patients concomitantly treated with cyclosporine the dosage and the trough-level of this agent were stable throughout the trial. In conclusion the present experience in paediatric outpatients with chronic kidney diseases supports the view that amlodipine is an effective and rather well tolerated antihypertensive drug when given once a day. PMID- 11439314 TI - Correlation of sodium-related factors with insulin sensitivity in young, lean, male offspring of hypertensive and normotensive subjects. AB - Pioneer studies have proposed that multiple metabolic abnormalities, such as insulin resistance, increased Na(+)-H(+) exchanger activity and abnormal intracellular calcium homeostasis, are frequently associated with a subset of essential hypertensive patients with low plasma renin activity (PRA). However, it is unclear whether insulin resistance is related to the low renin status in the very early phase of genetical hypertension. Besides, there is controversy on the subject of the in vivo effect of acute hyperinsulinaemia on sodium-related factors. We investigated the relationship between sodium-related parameters and insulin sensitivity, and the effects of euglycaemic hyperinsulinaemia on cyclic guanosine monophosphate (cGMP) and atrial natriuretic peptide (ANP) levels in 17 young, lean, normotensive male subjects, who displayed extreme predispositions for the development of hypertension. PRA was significantly lower in the positive than in the negative family history group (P < 0.05). Insulin sensitivity (M value) was correlated with PRA before euglycaemic hyperinsulinaemic clamping (r = 0.577, P < 0.05), and was also inversely correlated with fractional excretion of sodium (FE(Na)) before clamping (r = -0.51, P < 0.05). Euglycaemic hyperinsulinaemia significantly decreased PRA (P < 0.0001) and increased cGMP (P < 0.05) and ANP levels (P < 0.01). In conclusion, insulin sensitivity may be partially determined by PRA levels and FE(Na) before clamping in young, lean, normotensive male subjects. Acute euglycaemic hyperinsulinaemia decreases PRA, and increases cGMP and ANP levels from the fasting condition. PMID- 11439315 TI - Mild chronic renal insufficiency induces sympathetic overactivity. AB - The objectives of this study were to evaluate baseline sympathetic nerve activity as well as the mechanisms of sympathetic overactivity in mild chronic renal insufficiency hypertension. Seven hypertensives with mild renal insufficiency, seven hypertensives with normal renal function and seven normotensives, age and weight-matched were studied on one session to evaluate baseline muscle sympathetic nerve activity measured in the peroneal nerve. The mild renal insufficiency hypertensives and the hypertensives with normal renal function were also studied to evaluate arterial baroreflex control of muscle sympathetic nerve activity assessed by increasing and decreasing blood pressure through continuous infusion of phenylephrine and sodium nitroprusside respectively. Baseline muscle sympathetic nerve activity was significantly higher in mild renal insufficiency hypertensives (34 bursts/min) when compared to hypertensives with normal renal function (24 bursts/min) and to normotensives (16 bursts/min). Baroreflex control of muscle sympathetic nerve activity was (in absolute values) 15.2 in hypertensives with normal renal function vs 2.6 in mild renal insufficiency hypertensives (P < 0.05) during phenylephrine and 6.3 vs 8.2 during nitroprusside infusions. Mild renal insufficiency hypertensives showed sympathetic overactivity when compared to hypertensives with normal renal function and to normotensives. This finding demonstrates that elevated sympathetic activity may be precociously detected in renal insufficiency. Baroreflex gain to phenylephrine was blunted in mild renal insufficiency hypertensives when compared to hypertensives with normal renal function, suggesting that bradycardic response in mild renal insufficiency is blunted. PMID- 11439316 TI - Urinary albumin excretion in lean, overweight and obese glucose tolerant individuals: its relationship with dyslipidaemia, hyperinsulinaemia and blood pressure. AB - The presence of microalbuminuria has become an important tool for therapeutic intervention. In this study we investigated whether the dysmetabolic syndrome of obesity was associated with or could occur in the absence of microalbuminuria. The study was conducted in 71 clinically healthy, glucose tolerant Hispanics (age: 43 +/- 1.4 years, body mass index (BMI): 28.7 +/- 0.6 kg/m(2), systolic blood pressure (SBP): 117 +/- 2 mm Hg, diastolic blood pressure (DBP): 77 +/- 1.3 mm Hg, urinary albumin excretion: 10.2 +/- 0.6 mg/24 h). Subjects were classified as lean (BMI <25), overweight (BMI >25 <30) and obese (BMI >30 kg/m(2)). Greater BMI was associated with higher body weight, waist-to-hip ratio (WHR), BP, fasting insulin, triglyceride, post glucose load insulin and glucose, and lower high density lipoprotein (HDL) cholesterol levels. However, no significant differences in the urinary albumin excretion (mg/24 h) were found between lean (9.0 +/- 0.9; median: 9.1), overweight (11.3 +/- 1.2; median: 10.5) and obese (11.1 +/- 1.2; median: 9.7) subjects. In addition, microalbuminuria (urinary albumin excretion >30 mg/24 h) was not found in any of the study subjects. For all subjects combined, as well as for each of the groups separately, the urinary albumin excretion was unrelated to the BMI, WHR, body weight, triglyceride, cholesterol (total, LDL or HDL), fasting or post-load glucose and insulin plasma concentrations. Neither in females nor in males, abdominal fat accumulation was associated with an increase in the urinary albumin excretion. However, in the obese groups, urinary albumin excretion was strongly related to the level of SBP (r(2): 0.67; P < 0.0001) and DBP (r(2): 0.55; P < 0.0001). In summary, obesity, hyperinsulinaemia and dyslipidaemia per se are not determinants of increased albumin excretion. However, in the obese subjects, the BP, particularly the SBP, was a strong determinant of the level of albumin in the urine. Microalbuminuria may occur later in the course of the dysmetabolic syndrome, due to worsening of hypertension and development of hyperglycaemia. PMID- 11439317 TI - Prognostic significance of blood pressure measured on rising. AB - Previous works using ambulatory blood pressure (BP) monitoring demonstrated that independently of the mean level of BP, the variability in BP, or the day-night range, could have prognostic significance. We have also found that the value of BP on rising in the morning is strongly correlated with left ventricular mass of hypertensive individuals independently of the 24-h value. In the present study, we sought its predictive value for cardiovascular complications in a cohort of hypertensive patients. The population studied belongs to a cohort of initially untreated hypertensive patients recruited since 1983 and followed for more than 5 years. Patients were then treated and followed by their family doctor. At entry, all patients were equipped with a device to measure ambulatory BP. They were requested to trigger a measurement manually on rising in the morning (arising BP). The data on their outcome were collected by a physician unaware of the initial state of the patients. A total of 256 patients have been followed up for 5 years or more, 19 were lost to follow-up. The mean follow-up period was 84 +/- 29 months. Cardiovascular complications were recorded in 23 individuals. The arising systolic BP (SBP) was significantly higher in the group who presented a complication. In a stepwise discriminant analysis including age, office, fitting, arising and 24-h average SBPs only age and arising SBP entered the equation. In conclusion, the single BP value measured by an ambulatory device on rising in the morning seems more discriminant of future cardiovascular events than the value of BP measured on fitting the device or the average of three measurements taken under standardised conditions in the hospital or office. PMID- 11439318 TI - The measurement of blood pressure and the detection of hypertension in children and adolescents. AB - Despite the publication of several expert committee guidelines for the measurement of blood pressure (BP) and the diagnosis of hypertension in children and adolescents, it was our perception and clinical experience that there still appeared to be a general lack of standardisation of BP measurement techniques and little consensus on the criteria for diagnosing hypertension. To investigate this further, we have conducted a postal survey of consultant-grade paediatricians who were members of the British Paediatric Association (BPA). A total of 1500 questionnaires were sent out and 708 analysable replies were received (47.1%). This showed that 68.6% of paediatricians routinely measured BP, at least on one occasion, in children or adolescents attending their outpatient clinics, 17.7% started at or soon after birth, 12.3% started at the age of 1 year, 20.0% at 3 years, 12.0% from 7 years of age and 3.5% from the age of 13. Only 60.5% reported that they had a choice of four or more different cuff sizes in their clinic. Forty-one percent of respondents reported that the BP was always or sometimes measured by nurses. Fifty-one percent of respondents measured diastolic BP at the phase of muffling of sound (Korotkoff phase IV), 31.9% used the disappearance of sound (phase V) whilst 15.9% claimed that they measured both end-points. The criteria for diagnosing a child as being hypertensive varied greatly; 17.9% reported that they responded to the systolic BP alone, 13.5% to the diastolic BP alone, 65.9% relied on both pressures, and 2.7% responded to either the systolic or diastolic pressure if it was raised. Furthermore, 12.9% diagnosed hypertension if the BP exceeded the 90th percentile in relation to age and 41.8% used the 95th percentile. However 45.3% of respondents employed a higher dividing line. In hospitalised children, leg blood pressures were measured routinely by 30.3%, although a further 44.0% would do so if aortic coarctation or other vascular diseases were suspected. Despite considerable variation in clinical practice, techniques and criteria, only 11.4% of clinicians would manage the patients themselves, with the remainder referring the child on to the appropriate specialist. The survey suggests a general lack of standardisation of BP measurement techniques and little consensus on the criteria for diagnosing hypertension amongst paediatricians. Simplified, shortened and updated guidelines on hypertension in paediatric practice and research are needed. PMID- 11439319 TI - T594M and G442V polymorphisms of the sodium channel beta subunit and hypertension in a black population. AB - Polymorphisms of the epithelial sodium channel may raise blood pressure by increasing renal sodium reabsorption. This study examines frequency distributions and associations with hypertension of the T594M and of the G442V polymorphisms of the beta subunit of the epithelial sodium channel in a population-based sample. We studied a stratified random sample of 459 subjects (279 women), aged 40-59 years, of black African origin from general practices' lists within a defined area of South London. All were first generation immigrants. The polymorphic variants were detected using single strand conformational polymorphism technique (SSCP). The prevalence of hypertension (BP > or =160 and/or 95 mm Hg or on drug therapy) was 43%; of these, 76% were on drug therapy. The main analysis was carried out by three ordered blood pressure categories (I to III) according to increasing blood pressure and presence or absence of drug therapy. The frequency of the 594M variant (heterozygotes and homozygotes) was 4.6%; the frequency of the 442V variant was higher (27.0%). The frequency of the 594M variant increased with increasing blood pressure category (P = 0.05) and was more common in hypertensives than normotensives. By contrast the frequency of the 442V variant did not vary across increasing blood pressure categories (P = 0.62). No gender difference was observed. Adjustment for age, sex and body mass index did not alter these findings. These results suggest that the 594M variant may contribute to high blood pressure in black people of African origin whereas the G442V polymorphism is unlikely to influence blood pressure in this population. PMID- 11439320 TI - Regression of atherosclerotic renal artery stenosis with aggressive lipid lowering therapy. PMID- 11439321 TI - Neutral effect of valsartan on serum uric acid and renal function tests. PMID- 11439322 TI - Angiotensin receptor antagonists and vaso-vagal attacks due to sensitisation of the Bezold-Jarisch reflex? PMID- 11439323 TI - Differential post-translational modification of the tumour suppressor proteins Rb and p53 modulate the rates of radiation-induced apoptosis in vivo. AB - Ionizing radiation induces p53-dependent apoptosis in the spleen, providing a model system to study p53 regulated events in a normal cell type. We have developed an in vivo model that identifies genetic differences in the regulation of p53-mediated apoptosis and addresses whether altered post-translational events in the p53-p21/Rb axis modulate the sensitivity of cells to radiation-induced cell death in vivo. Splenocytes from mice with distinct genetic backgrounds (DBA/2 and C57BL/6) exhibit differences in the rate of apoptosis. Whilst no obvious strain differences in protein levels of Bcl-2 or the cyclin-CDKs were observed, early post-translational regulatory events in the p53-p21/Rb axis showed striking differences in the two mouse strains. Cells from C57BL/6 animals undergo more rapid apoptosis after irradiation resulting from elevated levels and rapid induction of p53, pronounced Rb-cleavage, and the absence of a sustained induction of p21. In contrast, cells from DBA/2 animals have a reduced rate of apoptosis following irradiation with elevated levels of hyperphosphorylated Rb and a sustained induction of the p21 protein that is coincident with the C terminal phosphorylation of p53. These data suggest that quantitative differences in the level of p21 protein can affect the rate of apoptosis in vivo, consistent with the view that p21 is an anti-apoptotic effector of p53. However, striking differences in the Rb protein-caspase cleavage or hyperphosphorylation-in the same cell type, but in different genetic backgrounds, demonstrates that p53 dependent apoptosis can be modulated in vivo by genetic factors that impinge upon the pro- or anti-apoptotic potential of Rb. In addition, we show that Rb cleavage is p53-dependent and that its phosphorylation status can be uncoupled from p21 expression. This study highlights the possibility that genetic factors can be identified that affect differential sensitivity of cells to ionizing radiation in vivo. PMID- 11439324 TI - Variations in Prkdc encoding the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) and susceptibility to radiation-induced apoptosis and lymphomagenesis. AB - DNA double-strand breaks (DSBs) induced by ionizing radiation enforce cells to die, if unrepaired; while if misrepaired, DSBs may cause malignant transformation. The DSB repair system predominant in mammals requires DNA dependent protein kinase (DNA-PK). Previously, we identified the apoptosis susceptibility gene Radiation-induced apoptosis 1 (Rapop1) on mouse chromosome 16. The STS/A (STS) allele at Rapop1 leads to decreased sensitivity to apoptosis in the BALB/cHeA (BALB/c) background. In the present study, we established Rapop1 congenic strains C.S-R1 and C.S-R1L, which contain the STS genome in a 0.45 cM interval critical for Rapop1 in common in the BALB/c background. Within the segment critical for Rapop1, Prkdc encoding the catalytic subunit of DNA-PK (DNA PKcs) was assigned. Two variations T6,418C and G11,530A, which induce amino acid substitutions C2,140R downstream from the putative leucine zipper motif and V3,844M near the kinase domain, respectively, were found between BALB/c and STS for Prkdc. The majority of inbred strains such as C57BL/6J carried the STS allele at Prkdc; a few strains including 129/SvJ and C.B17 carried the BALB/c allele. DNA-PK activity as well as DNA-PKcs expression was profoundly diminished in BALB/c and 129/SvJ mice as compared with C57BL/6 and C.S-R1 mice. In the crosses (C.S-R1 x BALB/c)F(1) x 129/SvJ and (C.S-R1 x BALB/c)F(1) x C.B17, enhanced apoptosis occurred in the absence of the wild-type allele at Prkdc. C.S-R1 and C.S-R1L were both less sensitive to radiation lymphomagenesis than BALB/c. Our study provides strong evidence for Prkdc as a candidate for Rapop1 and a susceptibility gene for radiation lymphomagenesis as well. PMID- 11439325 TI - Hepatitis B virus X mutants derived from human hepatocellular carcinoma retain the ability to abrogate p53-induced apoptosis. AB - Chronic hepatitis B virus (HBV) infection and the integration of its X gene (HBx) are closely associated with the development of hepatocellular carcinoma (HCC). The integrated HBx frequently is truncated or contains point mutations. Previous studies indicated that these HBx mutants have a diminished co-transactivational activity. We have compared the effects of wild-type (wt) HBx and its naturally occurring mutants derived from human HCCs on transcriptional co-transactivation, apoptosis and interactive effects with p53. We demonstrated that overexpression of mutant, but not wt HBx, is defective in transcriptional co-transactivation of the NF-kappaB-driven luciferase reporter. By using a microinjection technique, the HBx mutants were shown to have an attenuated pro-apoptotic activity. This deficiency may be attributed to multiple mutations in the co-transactivation domain of HBx, that leads to decreased stability of the translated product. However, wt or mutant HBx bind to p53 in vitro and retain their ability to block p53-mediated apoptosis in vivo, which has been implicated as its major tumor suppressor function. The abrogation of p53-mediated apoptosis by integrated HBx mutants may provide a selective clonal advantage for preneoplastic or neoplastic hepatocytes and contribute to hepatocellular carcinogenesis. PMID- 11439326 TI - The HPV E7 oncoprotein inhibits tumor necrosis factor alpha-mediated apoptosis in normal human fibroblasts. AB - Tumor necrosis factor-alpha (TNF) is a cytokine that induces programmed cell death, apoptosis, in a number of cell types and is employed by cytotoxic T cells to eliminate virus infected cells. Consequently, many viruses have acquired mechanisms to undermine these host cell defense mechanisms and cause resistance to TNF-mediated apoptosis. Here we show that normal human diploid fibroblasts that express the human papillomavirus type 16 E7 oncoprotein have a decreased propensity to undergo apoptosis in response to TNF treatment. The ability of E7 to undermine TNF-mediated apoptosis correlates with cellular transformation. While E7 does not generally subvert signaling by tumor necrosis factor receptor 1, pro-caspase 8 activation is decreased in E7-expressing cells. E7 also provides some protection from apoptosis caused by stimulation of the TNF receptor 1 related cytokine receptor Fas, where induction of apoptosis occurs much slower in this cell type. Hence, E7-expressing normal human fibroblasts exhibit a specific defect that obstructs cytokine-mediated activation of pro-caspase 8 and apoptosis. PMID- 11439327 TI - A splice variant of Skp2 is retained in the cytoplasm and fails to direct cyclin D1 ubiquitination in the uterine cancer cell line SK-UT. AB - Cyclin D1 is an important regulator of the transition from G1 into S phase of the cell cycle. The level to which cyclin D1 accumulates is tightly regulated. One mechanism contributing to the control of cyclin D1 levels is the regulation of its ubiquitination. SK-UT-1B cells are deficient in the degradation of D-type cyclins. We show here that p27, a substrate of the SCF(Skp2) ubiquitin ligase complex, is coordinately stabilized in SK-UT-1B cells. Further, we show that expression of Skp2 in SK-UT-1B cells rescues the cyclin D1 and p27 degradation defect observed in this cell line. These results therefore indicate that the SCF(Skp2) ubiquitin ligase complex affects the ubiquitination of cyclin D1. In addition, we show that SK-UT-1B cells express a novel splice variant of Skp2 that localizes to the cytoplasm and that cyclin D1 ubiquitination takes place in the nucleus. We propose that the translocation of Skp2 into the nucleus is required for the ubiquitination of cyclin D1 and that the absence of the SCF(Skp2) complex in the nucleus of SK-UT-1B cells is the mechanism underlying the ubiquitination defect observed in this cell line. Finally, our data indicates that differential splicing of F-box proteins may represent an additional level of regulation of the F-box mediated ubiquitination pathway. PMID- 11439328 TI - Leukemic transformation of normal murine erythroid progenitors: v- and c-ErbB act through signaling pathways activated by the EpoR and c-Kit in stress erythropoiesis. AB - Primary erythroid progenitors can be expanded by the synergistic action of erythropoietin (Epo), stem cell factor (SCF) and glucocorticoids. While Epo is required for erythropoiesis in general, glucocorticoids and SCF mainly contribute to stress erythropoiesis in hypoxic mice. This ability of normal erythroid progenitors to undergo expansion under stress conditions is targeted by the avian erythroblastosis virus (AEV), harboring the oncogenes v-ErbB and v-ErbA. We investigated the signaling pathways required for progenitor expansion under stress conditions and in leukemic transformation. Immortal strains of erythroid progenitors, able to undergo normal, terminal differentiation under appropriate conditions, were established from fetal livers of p53-/- mice. Expression and activation of the EGF-receptor (HER-1/c-ErbB) or its mutated oncogenic version (v ErbB) in these cells abrogated the requirement for Epo and SCF in expansion of these progenitors and blocked terminal differentiation. Upon inhibition of ErbB function, differentiation into erythrocytes occurred. Signal transducing molecules important for renewal induction, i.e. Stat5- and phosphoinositide 3 kinase (PI3K), are utilized by both EpoR/c-Kit and v/c-ErbB. However, while v ErbB transformed cells and normal progenitors depended on PI3K signaling for renewal, c-ErbB also induces progenitor expansion by PI3K-independent mechanisms. PMID- 11439329 TI - Down-regulation of cathepsin B expression impairs the invasive and tumorigenic potential of human glioblastoma cells. AB - Increases in abundance of cathepsin B transcript and protein correlate with increases in tumor grade and alterations in subcellular localization and activity of cathepsin B. The enzyme is able to degrade the components of the extracellular matrix (ECM) and activate other proteases capable of degrading ECM. To investigate the role played by this protease in the invasion of brain tumor cells, we transfected SNB19 human glioblastoma cells with a plasmid containing cathepsin B cDNA in antisense orientation. Control cells were transfected with vector alone. Clones expressing antisense cathepsin B cDNA exhibited significant reductions in cathepsin B mRNA, enzyme activity and protein compared to controls. Matrigel Invasion assay showed that the antisense-transfected cells had a markedly diminished invasiveness compared with controls. When tumor spheroids containing antisense transfected SNB19 cells expressing reduced cathepsin B were co-cultured with fetal rat brain aggregates, invasion of fetal rat brain aggregates was significantly reduced. Green Fluorescent Protein (GFP) expressing parental cells and antisense transfectants were generated for detection in mouse brain tissue without any post-chemical treatment. Intracerebral injection of SNB19 stable antisense transfectants resulted in reduced tumor formation in nude mice. These results strongly support a role for cathepsin B in the invasiveness of human glioblastoma cells and suggest cathepsin B antisense may prove useful in cancer therapy. PMID- 11439330 TI - Distinctive gene expression profiles associated with Hepatitis B virus x protein. AB - Hepatitis B virus (HBV) is a major risk factor for the development of hepatocellular carcinoma (HCC). HBV encodes the potentially oncogenic HBx protein, which mainly functions as a transcriptional co-activator involving in multiple gene deregulations. However, mechanisms underlying HBx-mediated oncogenicity remain unclear. To determine the role(s) of HBx in the early genesis of HCC, we utilized the NCI Oncochip microarray that contains 2208 human cDNA clones to examine the gene expression profiles in either freshly isolated normal primary adult human hepatocytes (Hhep) or an HCC cell line (SK-Hep-1) ecotopically expressing HBx via an adenoviral system. The gene expression profiles also were determined in liver samples from HBV-infected chronic active hepatitis patients when compared with normal liver samples. The microarray results were validated through Northern blot analysis of the expression of selected genes. Using reciprocally labeling hybridizations, scatterplot analysis of gene expression ratios in human primary hepatocytes expressing HBx demonstrates that microarrays are highly reproducible. The comparison of gene expression profiles between HBx-expressing primary hepatocytes and HBV-infected liver samples shows a consistent alteration of many cellular genes including a subset of oncogenes (such as c-myc and c-myb) and tumor suppressor genes (such as APC, p53, WAF1 and WT1). Furthermore, clustering algorithm analysis showed distinctive gene expression profiles in Hhep and SK-Hep-1 cells. Our findings are consistent with the hypothesis that the deregulation of cellular genes by oncogenic HBx may be an early event that favors hepatocyte proliferation during liver carcinogenesis. PMID- 11439331 TI - Modulation in vitro of H-ras oncogene expression by trans-splicing. AB - In man, activated N-, K- and H-ras oncogenes have been found in around 30% of the solid tumours tested. An exon known as IDX, which has been described previously and is located between exon 3 and exon 4A of the c-H-ras pre-mRNA, allows an alternative splicing process that results in the synthesis of the mRNA of a putative protein named p19. It has been suggested that this alternative pathway is less tumorigenic than that which results in the activation of p21. We have used the mammalian trans-splicing mechanism as a tool with which to modulate this particular pre-mRNA processing to produce mRNA similar to that of mature p19 RNA. The E4A exon of the activated H-ras gene was found to be a good target for external trans-splicing. We reprogrammed the rat carnitine octanoyltransferase exon 2 to specifically invade the terminal region of H-ras. Assays performed with this reprogrammed trans-exon showed that the trans-splicing product was obtained in competition with cis-splicing of the D intron of the H-ras gene, and was associated with concomitant down-modulation of D intron cis-splicing. We also found that the exon 4A of the human c-H-ras gene underwent successive trans splicing rounds with an external exon. PMID- 11439332 TI - p27 Kip1 inhibits HER2/neu-mediated cell growth and tumorigenesis. AB - HER2/neu, a receptor tyrosine kinase oncogene, promotes mitogenic growth and transformation of cancer cells. We previously identified that its oncogenic signals down-regulate the cyclin-dependent kinase inhibitor p27 Kip1, which is defined as a haplo-insufficient tumor suppressor. Here, we applied the human p27 gene as a novel anticancer agent for HER2/neu-overexpressing cells under the control of a tetracycline (tet)-regulated gene expression system. Overexpression of p27 inhibits HER2/neu-activated CDK2 activity, cell proliferation, and transformation. Most significantly for clinical application, p27 expression in HER2/neu-overexpressing cells can be regulated in vivo and reduce the tumor volume in a tumor model. The findings demonstrate the applicability of employing p27 in HER2/neu-associated cancer gene therapy. PMID- 11439333 TI - Thioredoxin participates in a cell death pathway induced by interferon and retinoid combination. AB - Interferons (IFNs) and retinoids are potent tumor growth suppressors. We have shown earlier that the IFN-beta and all-trans retinoic acid combination, but not the single agents, induces death in several tumor cell lines. Employing a genetic approach we have recently identified several Genes associated with Retinoid-IFN induced Mortality (GRIM) that mediate the cell death effect of IFN/RA combination. One of the GRIMs, GRIM-12, was identical to human thioredoxin reductase (TR), an enzyme that controls intracellular redox state. To define the participants of TR mediated death pathway we have examined the role of thioredoxin (Trx), its downstream substrate, and its influence on IFN/RA-induced death regulation. Inhibition of the thioredoxin expression by antisense RNA suppressed cell death. Similarly, a mutant Trx1 lacking the critical cysteine residues blocked cell death. In contrast, overexpression of wildtype thioredoxin augmented cell death. This effect of Trx1 was in part due to its ability to augment cell death via caspase-8. The redox inactive Trx1 mutant inhibits the cell death induced by caspase-8 but not caspase-3. These studies identify a novel mechanism of cell death regulation by IFN/RA combination involving redox enzymes. PMID- 11439334 TI - TEL contacts multiple co-repressors and specifically associates with histone deacetylase-3. AB - TEL (Translocation-ETS-Leukemia or ETV 6) is disrupted by multiple chromosomal translocations in acute leukemia. The loss of heterozygosity at the TEL locus in leukemias and the hemizygous deletion of TEL that is observed in various tumors, suggests that TEL is a tumor suppressor. Overexpression of TEL alters cellular morphology and represses the expression of the matrix metalloproteinase stromelysin-1. Based on these studies, deletion analysis was used to define the minimal repression domains of TEL. TEL-mediated repression required both the N terminal pointed domain and a central region composed of amino acids 268-303. The mSin3A and N-CoR corepressors bind to the pointed domain and the central repression domain of TEL, respectively. Unexpectedly, histone deacetylase-3, but not other histone deacetylases, also associates with the central region of TEL. Histone deacetylase-3 interacts with a TEL mutant that cannot bind N-CoR, suggesting that this is a direct interaction with TEL. In addition, histone H3 was under-acetylated near the TEL-binding sites in the endogenous stromelysin-1 promoter when TEL was expressed. Furthermore, trichostatin A, a potent histone deacetylase inhibitor, impaired TEL-dependent repression of the stromelysin-1 promoter. Finally, while TEL-expression induced cellular aggregation of Ras transformed cells, Trichostatin A reversed the TEL-induced cellular aggregation phenotype. Thus, the cumulative data suggests that histone deacetylase-3 activity is required for the transcriptional functions of TEL. PMID- 11439335 TI - The monoclonal antibody 225 activates caspase-8 and induces apoptosis through a tumor necrosis factor receptor family-independent pathway. AB - We previously reported that the anti-epidermal growth factor (EGF) receptor monoclonal antibody (mAb) 225 induces DiFi colon cancer cells to undergo apoptosis, and this apoptosis was accompanied by activation of the two apoptosis initiation caspases, caspase-8 and caspase-9. In the current study, we found that pretreatment of DiFi cells with the caspase-8-specific inhibitor z-IETD-fmk but not pretreatment with the caspase-9-specific inhibitor z-LEHD-fmk inhibited mAb 225-induced apoptosis, indicating that caspase-8 plays an essential role in initiating mAb 225-induced apoptosis. Because caspase-8 is activated primarily by the members of the tumor necrosis factor (TNF) receptor family, such as Fas, TNF receptor-1 (TNFR1), or receptors for TNF-related apoptosis-inducing ligand (TRAIL), we investigated whether mAb 225 activated caspase-8 by regulating one or more of these known pathways. Exposure of DiFi cells to TNFalpha or TRAIL activated caspase-8 and induced apoptosis in the cells. A TNFR1-antagonistic mAb or a TRAIL decoy receptor inhibited the activation of caspase-8 and the subsequent apoptosis induced by TNFalpha or TRAIL, respectively, in the cells. However, neither the TNFR1-antagonistic mAb nor the TRAIL decoy receptor inhibited mAb 225-induced activation of caspase-8 and apoptosis in DiFi cells. DiFi cells express detectable level of Fas but are not sensitive to the treatment by the Fas-agonistic mAb CH-11. A Fas-antagonistic mAb (ZB-4) inhibited the Fas agonistic mAb CH-11-induced caspase-8 activation and apoptosis in Jurkat T leukemic cells (used as positive control), but had no effect on mAb 225-induced activation of caspase-8 and apoptosis in DiFi cells. Taken together, our results suggest that mAb 225 does not interact with or regulate these known death receptor pathways. An exploration is therefore warranted for a novel mechanism by which mAb 225 activates caspase-8 and triggers apoptosis in DiFi cells. PMID- 11439336 TI - Hepatocyte Growth Factor/scatter factor-induces phosphorylation of cortactin in A431 cells in a Src kinase-independent manner. AB - The Hepatocyte Growth Factor receptor transduces proliferating and scattering signals in epithelial and endothelial cells. We have explored potential interactions of the HGF/SF receptor beta-subunit (p145(beta MET)) with F-actin binding partners aiming to identify novel downstream effectors implicated in HGF/SF pluripotent signalling. Cortactin, a p80/85 F-actin binding protein, was found phosphorylated on tyrosine in response to HGF-SF in A431 human epidermoid carcinoma cells, expressing the HGF/SF receptor (c-MET). The HGF/SF receptor was enriched in the detergent-insoluble fraction and was found to co-precipitate with cortactin and to associate in vitro with cortactin. The Grb2 small adapter protein known to associate via its Src homology 2 domain (SH2) with the MET C terminus, was also associated with cortactin. Transient transfection of A431 cells with dominant-negative Grb2 constructs has revealed that the Grb2-C-SH3 domain possesses a central role in cortactin phosphorylation in response to HGF/SF. Finally, tyrosine phosphorylation of cortactin was found uncoupled of endogenous c-Src kinase activity, thus further supporting the hypothesis that cortactin is a direct target of the MET kinase. We propose that cortactin may constitute a docking site for MET-derived signals within the cytoskeleton. PMID- 11439337 TI - Myc drives apoptosis in PC12 cells in the absence of Max. AB - A conditionally active chimeric form of the c-Myc protein fused to the ligand binding domain of the estrogen receptor (MycER) was expressed in PC12 cells. Induction of Myc activity resulted in a threefold increase in apoptosis after 5 days when cells were maintained in 1% serum. The effect of Myc overexpression was dependent on its DNA-binding domain but not on its heterodimeric binding protein Max, which is absent in PC12 cells. Preincubation of the c-Myc overexpressing cells with either NGF or bFGF, but not EGF, prevented the Myc-mediated increase in apoptosis, although the signaling pathways used by NGF and bFGF to block cell death differed. NGF-mediated rescue was mediated by the phosphatidylinositol 3' OH (P13) kinase/Akt pathway while rescue by bFGF was not affected by P13 kinase inhibitors. These results show that Myc can induce apoptosis in PC12 cells in a Max-independent manner and that alternate signaling pathways exist to mediate cell survival. PMID- 11439338 TI - Regulation of vascular endothelial growth factor expression by acidosis in human cancer cells. AB - The influence of acidosis on the expression of the vascular endothelial growth factor (VEGF) gene was determined. FG human pancreatic adenocarcinoma cells were incubated for various time periods in media at a physiologically relevant pH level (6.7-7.4). The expression of VEGF mRNA and protein secretion was inversely correlated with pH in a pH- and time-dependent manner. Transient acidosis also activated the VEGF promoter/enhancer luciferase reporter, which was consistent with an increased VEGF gene transcription rate and VEGF mRNA half-life. These data indicated that acidosis transcriptionally and posttranscriptionally regulates VEGF expression, suggesting that an acidic tumor microenvironment contributes to tumor angiogenesis and progression. PMID- 11439339 TI - TRAIL (APO-2L) induces apoptosis in human prostate cancer cells that is inhibitable by Bcl-2. AB - To determine if TRAIL-induced apoptosis in human prostate tumor cells was suppressed by bcl-2, we compared the levels of apoptosis induced by recombinant human TRAIL in pairs of isogenic cell lines that do or do not express bcl-2. Three human prostate tumor cell lines (PC3, DU145 and LNCaP) and their bcl-2 expressing counterparts were tested for their susceptibility to TRAIL. Cells were exposed to TRAIL in the presence of cycloheximide which acted as a sensitizer. Apoptosis was induced rapidly in PC3 and DU145 neo-control transfected cells, whereas induction in LNCaP required 24 h. All three cell line variants expressing bcl-2 were resistant to the apoptotic effects of TRAIL. Caspase 3 and 8 activation was also detected in the neo control cells after treatment with TRAIL, demonstrating the rapid activation of the caspase cascade similar to that seen with other death receptors. Bcl-2 overexpression in these cells blocked activation of these caspases, suggesting that bcl-2 expression of human cancer cells may be a critical factor in the therapeutic efficacy of TRAIL. PMID- 11439340 TI - Regulation of the cell cycle by p53 after DNA damage in an amphibian cell line. AB - In mammalian cells, the p53 protein is a key regulator of the cell cycle following DNA damage. In the present study, we investigated the function of p53 in the A6 amphibian cell line. Using various specific Xenopus p53 monoclonal antibodies, we showed that Xenopus p53 accumulates after DNA damage, including gamma and UV irradiation or treatment with adriamycin. Such accumulation is accompanied by an increase in the apparent molecular weight of the protein. This change was shown to be the result of a phosphorylation event that occurs after DNA damage. Accumulation of Xenopus p53 is parallel to a drastic change in the cell cycle distribution. Brief exposure to adriamycin or gamma irradiation induces reversible growth arrest, whereas long-term exposure to adriamycin leads to apoptosis. Taken together, these results indicate that p53 has a similar behaviour in frog cells and mammalian cells, and that it conserves two activities, cell cycle arrest and apoptosis. PMID- 11439341 TI - Site-specific and temporally-regulated retinoblastoma protein dephosphorylation by protein phosphatase type 1. AB - pRb is dephosphorylated at mitotic exit by the type 1 serine/threonine protein phosphatases (PP1). Here we demonstrate for the first time that mitotic pRb dephosphorylation is a sequential, temporally-regulated event. We also provide evidence that the three mammalian isoforms of PP1, alpha, gamma-1, and delta, differ in their respective preferences for site-specific pRb dephosphorylation and that the mitotic and G(1) PP1-isoform counterparts exhibit differential activities towards mitotic pRb. Finally, the physiological relevance of the striking contrast between the patterns of Thr821 and Thr826 dephosphorylation, sites known to be important for disrupting binding of LXCXE-containing proteins to pRb, is addressed. PMID- 11439342 TI - Anti-mitotic properties of indirubin-3'-monoxime, a CDK/GSK-3 inhibitor: induction of endoreplication following prophase arrest. AB - The bis-indole indirubin is the active ingredient of the Traditional Chinese Medicine recipe Danggui Longhui Wan used against chronic myelocytic leukemia. We have previously shown that indirubins are potent inhibitors of cyclin-dependent kinases and glycogen synthase kinase-3. We here investigated the anti-mitotic properties of this class of compounds using the cell permeable indirubin-3' monoxime and the HBL-100 cell line. Indirubin-3'-monoxime reversibly arrests asynchronous HBL-100 cells in G2. This arrest is not accompanied by any significant change in expression of the major cell cycle regulators. However indirubin-3'-monoxime inhibits the phosphorylation of consensus CDK phosphorylation sites as well as of nucleolin at a specific CDK1/cyclin B phosphorylation site, suggesting a direct action on the mitotic CDK1/cyclin B. When indirubin-3'-monoxime is added to HBL-100 cells synchronized in M phase by nocodazole, cells undergo an endoreplication leading to an 8n DNA content. As soon as indirubin-3'-monoxime is washed away, these polyploid cells become aneuploid and later die from necrosis. This mechanism of endoreplication followed by cell death may contribute to the anti-tumour properties of indirubins. PMID- 11439343 TI - The polycomb protein MPc3 interacts with AF9, an MLL fusion partner in t(9;11)(p22;q23) acute leukemias. AB - Polycomb group (PcG) proteins assemble to form large multiprotein complexes involved in gene silencing. Evidence suggests that PcG complexes are heterogeneous with respect to both protein composition and specific function. MPc3 is a recently described mouse Polycomb (Pc) protein that shares structural homology with at least two other Pc proteins, M33 and MPc2. All three Pc proteins bind another PcG protein, RING1, through a conserved carboxy-terminal C-box motif. Here, data are presented demonstrating that MPc3 also interacts with AF9, a transcriptional activator implicated in the development of acute leukemias. The carboxy-terminus of AF9 is fused to the MLL protein in leukemias characterized by t(9;11)(p22;q23) chromosomal translocations. Importantly, it is the carboxy terminus of AF9 to which MPc3 binds. The AF9 binding site of MPc3 maps to a central, non-conserved, region of the polypeptide sequence. In contrast to MPc3, data indicate that the Pc protein M33 does not interact with AF9. This finding suggests a potentially unique role for MPc3 in linking a PcG silencing complex to a transcriptional activator protein. PMID- 11439344 TI - Low concentrations of paclitaxel induce cell type-dependent p53, p21 and G1/G2 arrest instead of mitotic arrest: molecular determinants of paclitaxel-induced cytotoxicity. AB - Paclitaxel (PTX), a microtubule-active agent, blocks cell proliferation by inhibiting mitotic progression leading to mitotic and postmitotic arrest and cell death. Here we demonstrate for the first time that very low concentrations of PTX (3-6 nM) can completely inhibit cell proliferation without arresting cells at mitosis. At these low concentrations that are insufficient to inhibit mitotic progression, PTX induced both p53 and p21 causing G1 and G2 arrest in A549. In contrast, low PTX concentrations failed to induce G1 and G2 arrest in A549/E6 cells, that do not express p53. Furthermore, we observed that the levels of p53 and p21 induced by adriamycin and by low concentrations of PTX in A549 cells were comparable. This observation led us to conclude that low concentrations of PTX can induce p53 and p21 sufficiently to cause G1 and G2. Many other cell lines, including HCT116 cells, do not readily upregulate p53 in response to PTX, and therefore undergo exclusively mitotic and postmitotic arrest after PTX treatment. At low concentrations that do not cause mitotic arrest, PTX did not significantly inhibit proliferation of these cells. In HCT116 cells, loss of p53 (HCT/p53(-/-)) or p21 (HCT/p21(-/-)) affects both Bax and Bcl-2 expression. In cells lacking p53, levels of Bax and p21 were decreased. In cells lacking p21, levels of wt p53 were highly increased to compensate for the loss of p21. This in turn results in upregulation of Bax and downregulation of Bcl-2 resulting in an increase of the apoptotic Bax/Bcl2 ratio consistent with increased sensitivity of these cells to apoptotic stimuli. High levels of p53 and Bax/Bcl-2 ratio can also explain why loss of p21 is rarely found in human cancer. PMID- 11439345 TI - Mechanisms of inactivation of E-cadherin in breast carcinoma: modification of the two-hit hypothesis of tumor suppressor gene. AB - Loss of heterozygosity (LOH) allows the expression of recessive mutation in tumor suppressor genes (TSG). Therefore, on the basis of Knudson's 'two-hit' hypothesis for TSG inactivation, the detection of a high LOH frequency in a chromosomal region is considered critical for TSG localization. One of these LOH regions in breast cancer is 16q22.1, which has been suggested to reflect the involvement of E-cadherin (E-cad), a cell-cell adhesion molecule. To confirm the tumorigenic role of E-cad, 81 sporadic invasive ductal carcinomas (IDCs) of the breast were tested for the 'two hits' required to inactivate this gene. A high frequency (37.3%) of LOH was detected in 67 informative tumors, but no mutation was found. To examine the possibility that transcriptional mechanisms serve as the second hit in tumors with LOH, specific pathways, including genetic variant and hypermethylation at the promoter region and abnormal expression of positive (WT1) and negative (Snail) transcription factors, were identified. Of these, promoter hypermethylation and increased expression of Snail were found to be common (>35%), and to be strongly associated with reduced/negative E-cad expression (P<0.05). However, unexpectedly, a significantly negative association was found between the existence of LOH and promoter hypermethylation (P<0.05), which contradicts the 'two-hit' model. Instead, since they coexisted in a high frequency of tumors, hypermethylation may work in concert with increased Snail to inactivate E-cad expression. Given that E-cad is involved in diverse mechanisms, loss of which is beneficial for tumors to invade but may also trigger apoptosis, this study suggests that maintaining a reversible mechanism, either by controlling the gene at the transcriptional level or by retaining an intact allele subsequent to LOH, might be important for E-cad in IDC and may also be common in TSGs possessing diverse functions. These findings provide clues to explain why certain TSGs identified by LOH cannot fulfil the two-hit hypothesis. PMID- 11439346 TI - Identification of the transmembrane dimer interface of the bovine papillomavirus E5 protein. AB - We have developed a genetic method to determine the active orientation of dimeric transmembrane protein helices. The bovine papillomavirus E5 protein, a 44-amino acid homodimeric protein that appears to traverse membranes as a left-handed coiled-coil, transforms fibroblasts by binding and activating the platelet derived growth factor (PDGF) beta receptor. A heterologous dimerization domain was used to force E5 monomers to adopt all seven possible symmetric coiled-coil registries relative to one another within the dimer. Focus formation assays demonstrated that dimerization of the E5 protein is required for transformation and identified a single preferred orientation of the monomers. The essential glutamine residue at position 17 resided in the dimer interface in this active orientation. The active chimera formed complexes with the PDGF beta receptor and induced receptor tyrosine phosphorylation. We also identified E5-like structures that underwent non-productive interactions with the receptor. PMID- 11439347 TI - Alternative lengthening of telomeres is associated with chromosomal instability in osteosarcomas. AB - Telomere maintenance is regarded as a key mechanism in overcoming cellular senescence in tumor cells and in most cases is achieved by the activation of telomerase. However there is at least one alternative mechanism of telomere lengthening (ALT) which is characterized by heterogeneous and elongated telomeres in the absence of telomerase activity (TA). We evaluated the prevalence of TA, gene expression of telomerase subunits and ALT in relation to telomere morphology and function in matrix producing bone tumors and in osteosarcoma cell lines and present evidence of a direct association of ALT with telomere dysfunction and chromosomal instability. Telomere fluorescence in situ hybridization (T-FISH) in ALT cells revealed elongated and shortened telomeres, partly in unusual configurations and loci, dicentric marker chromosomes and signal-free chromosome ends. Free ends give rise to end-to-end associations and may induce breakage fusion-bridge cycles resulting in an increased number of complex chromosomal rearrangements, as detected by multiplex-FISH (M-FISH). We propose that ALT cannot be seen as an equivalent to telomerase activity in telomere maintenance. Its association with telomere dysfunction and chromosomal instability may have major implications for tumor progression. PMID- 11439348 TI - Tyrosine kinase inhibitor STI571 enhances thyroid cancer cell motile response to Hepatocyte Growth Factor. AB - The Hepatocyte Growth Factor (HGF) and its receptor Met are physiological regulators of cell migration. HGF and Met have also been implicated in tumor progression and metastasis. We show here that the tyrosine kinase inhibitor STI571 has a stimulatory effect on HGF-induced migration and branching morphogenesis in thyroid cancer but not in primary or immortalized thyroid epithelial cells. These stimulatory effects of STI571 are observed at a concentration that is clinically relevant. The STI571-enhanced motile response can be correlated with an increase in the Met receptor tyrosine phosphorylation as well as ERK and Akt activation by HGF. Interestingly, one of the targets of STI571, namely the c-Abl tyrosine kinase, is activated by HGF and is recruited at the migrating edge of thyroid cancer cells. These data suggests that c-Abl and/or STI571-inhibited tyrosine kinases can negatively regulate the Met receptor to restrain the motile response in thyroid cancer cells. PMID- 11439349 TI - Activation of the IGF-IR system contributes to malignant growth of human and mouse medulloblastomas. AB - Insulin-like growth factor I receptor (IGF-IR) has been implicated in the normal and malignant growth of many cell types including cells from the central nervous system. In the cerebellar cortex IGF-IR mRNA is found in granular cells and IGF-I stimulation is mitogenic and protects cells from low-potassium-induced apoptosis. Since primitive neuroectodermal tumors/medulloblastomas (PNETs/medulloblastomas) are suspected to originate from the external cerebellar granular layer, it is reasonable to postulate that IGF-IR and/or its signaling molecules may contribute to the transformation of these poorly differentiated cells. To study activation of the IGF-IR system in medulloblastomas, we have utilized an antibody (anti pY1316) that specifically recognizes the phosphorylated (active) form of the IGF IR. Medulloblastoma biopsy specimens were positive when examined immunohistochemically with anti-Y1316 antibody. Further analysis of the IGF-IR system was performed in three human (Daoy, TE-671, D283 Med) and four mouse (BsB8, BsB13, Bs-1b, Bs-1c) medulloblastoma cell lines. All the murine cell lines examined express IGF-IR and PI3-kinase at relatively normal levels, and grossly overexpress IRS-1, when compared with normal mouse cerebellum. Within 15 min following IGF-I stimulation both mouse and human cell lines phosphorylate the beta subunit of the IGF-IR, IRS-1, Akt, and MAP kinases. They respond with cell proliferation when stimulated solely with IGF-I and are strongly inhibited when challenged with a dominant negative mutant of the IGF-IR (486/STOP), or with antisense oligonucleotides against the IGF-IR mRNA. PMID- 11439350 TI - Ras-GAP SH3 domain binding protein (G3BP) is a modulator of USP10, a novel human ubiquitin specific protease. AB - Degradation of cellular proteins through ubiquitination is a fundamental strategy for regulating biological pathways. De-ubiquitination, i.e. the removal of ubiquitin from proteins and peptides to which ubiquitin is attached, is catalyzed by processing proteases known as de-ubiquitinating enzymes. We are studying the biology of a family of de-ubiquitinating enzymes, the mammalian ubiquitin specific proteases (USPs), some of which appear to play a role in growth control. Given the fact that the modes of regulation of USPs and of their substrate specificity are poorly understood, we decided to attempt the identification of USP interacting proteins. Using the yeast two-hybrid system (2HS), we have isolated a cDNA clone whose product specifically interacts with USP10 but not with other USP baits tested. The isolated clone encodes a protein known to interact with the Ras-GTPase activating protein (G3BP). This interaction was further confirmed by performing a 2HS with G3BP, which led to the isolation of USP10 encoding cDNAs. We validated the interaction between the two proteins by performing in vitro binding assays and immunoprecipitations in human cells. G3BP does not appear to be a substrate of USP10; it rather inhibits the ability of USP10 to disassemble ubiquitin chains. The USP10/G3BP complex appears to co immunoprecipitate with ubiquitinated species that could be substrates of USP10. PMID- 11439351 TI - Distinct effects of PIAS proteins on androgen-mediated gene activation in prostate cancer cells. AB - Androgen signaling influences the development and growth of prostate carcinoma. The transcriptional activity of androgen receptor (AR) is regulated by positive or negative transcriptional cofactors. We report here that PIAS1, PIAS3, and PIASy of the protein inhibitor of activated STAT (PIAS) family, which are expressed in human prostate, display distinct effects on AR-mediated gene activation in prostate cancer cells. While PIAS1 and PIAS3 enhance the transcriptional activity of AR, PIASy acts as a potent inhibitor of AR in prostate cancer cells. The effects of PIAS proteins on AR are competitive. PIASy binds to AR but does not affect the DNA binding activity of AR. An NH2-terminal LXXLL signature motif of PIASy, although not required for PIASy-AR interaction, is essential for the transrepression activity of PIASy. Our results identify PIASy as a transcriptional corepressor of AR and suggest that different PIAS proteins have distinct effects on AR signaling in prostate cancer cells. PMID- 11439352 TI - Constitutive activation of nuclear factor-kappaB prevents TRAIL-induced apoptosis in renal cancer cells. AB - TRAIL has gained much attention for its specific induction of apoptosis in cancer cells but not in normal cells. This phenomenon has been explained thus: that cancer cells dominantly express death receptors while normal cells express decoy receptors. However, recent reports have shown that some cancer cell lines are resistant to TRAIL-induced apoptosis despite the absence of decoy receptors and the presence of death receptors. This suggested the existance of an inhibitory factor. We herein showed that NF-kappaB is a key molecule underlying the TRAIL resistant mechanism in renal cell carcinoma (RCC) cell lines. We observed that NF kappaB is constitutively activated in resistant cell lines. Forced expression of antisense cDNA of IkappaBalpha, a specific inhibitor of NF-kappaB, in TRAIL sensitive cell lines with a low NF-kappaB activity result in constitutive activation of NF-kappaB and resistance to TRAIL-induced apoptosis. Adenoviral expression of a stable form of IkappaBalpha in the TRAIL-resistant cell lines induced apoptosis. These data suggest that RCC can be classified into two subsets: TRAIL-sensitive RCC with a low NF-kappaB activity and TRAIL-resistant RCC with constitutively activated NF-kappaB. In the former group TRAIL can be a treatment option, while in the latter group a molecular approach targeting NF kappaB appears to be a promising therapy. PMID- 11439353 TI - The DNA binding activity of TAL-1 is not required to induce leukemia/lymphoma in mice. AB - Activation of the basic helix-loop-helix (bHLH) gene TAL-1 (or SCL) is the most frequent gain-of-function mutation in pediatric T cell acute lymphoblastic leukemia (T-ALL). Similarly, mis-expression of tal-1 in the thymus of transgenic mice results in the development of clonal T cell lymphoblastic leukemia. To determine the mechanism(s) of tal-1-induced leukemogenesis, we created transgenic mice expressing a DNA binding mutant of tal-1. Surprisingly, these mice develop disease, demonstrating that the DNA binding properties of tal-1 are not required to induce leukemia/lymphoma in mice. However, wild type tal-1 and the DNA binding mutant both form stable complexes with E2A proteins. In addition, tal-1 stimulates differentiation of CD8-single positive thymocytes but inhibits the development of CD4-single positive cells: effects also observed in E2A-deficient mice. Our study suggests that the bHLH protein tal-1 contributes to leukemia by interfering with E2A protein function(s). PMID- 11439354 TI - Functional characterization and role of INrf2 in antioxidant response element mediated expression and antioxidant induction of NAD(P)H:quinone oxidoreductase1 gene. AB - Antioxidant response element (ARE) and nuclear transcription factor Nrf2 are known to regulate expression and coordinated induction of NQO1 and other detoxifying enzyme genes in response to antioxidants and xenobiotics. A cytosolic inhibitor of Nrf2, INrf2, that retains Nrf2 in the cytoplasm, was cloned and sequenced. Treatment of cells with antioxidants and xenobiotics results in the release of Nrf2 from INrf2. Nrf2 then moves in the nucleus. This leads to the induction of ARE-mediated NQO1 and other detoxifying enzyme genes expression. INrf2 after dissociation from Nrf2 remains in the cytosol. Overexpression of INrf2 repressed ARE-mediated NQO1 gene expression. Deletion mapping of INrf2 revealed the requirement of KELCH domain (amino acid residues 361-597) and C terminal region (amino acid residues 598-624) in retention of Nrf2 in the cytosol. Both these regions of INrf2 independently retained Nrf2 in the cytosol leading to the repression of ARE-mediated NQO1 gene expression. These results may indicate that two different regions of INrf2 interact with a single molecule of Nrf2 or two or more molecules of Nrf2 interact with a single molecule of INrf2. The transcription of Nrf2 and INrf2 did not change in response to antioxidants and xenobiotics. This indicated that INrf2 and/or Nrf2 might be post transcriptionally modified in response to antioxidants and xenobiotics leading to the release of Nrf2 from INrf2 and induction of ARE-mediated NQO1 and other detoxifying enzyme genes expression. PMID- 11439355 TI - Heregulin-dependent translocation and hyperphosphorylation of ErbB-2. AB - Previous data have shown that in several tumor cells lines the addition of heregulin results in the translocation of ErbB-4 to a detergent-insoluble membrane fraction where it is hypertyrosine phosphorylated. The data herein demonstrate that heregulin or betacellulin, but not EGF, promotes the rapid translocation of ErbB-2, the heterodimerization partner for ErbB-4, to the same detergent-insoluble fraction in T47 D mammary carcinoma cells. The translocation of ErbB-2 and ErbB-4, but not ErbB-3, to this detergent-insoluble fraction is readily detected 2 min after the addition of heregulin. ErbB-2 present in this detergent-insoluble membrane fraction is tyrosine phosphorylated to a level fourfold greater than the majority of cellular ErbB-2 present in the detergent soluble membrane fraction. These results suggest the ligand-dependent formation of hyperphosphorylated ErbB-4/ErbB-2 dimers coordinate with translocation to a putative membrane microdomain. PMID- 11439356 TI - Role of p38 MAP kinases and ERK in mediating ultraviolet-B induced cyclooxygenase 2 gene expression in human keratinocytes. AB - The roles of p38 MAP kinases and ERK in UVB induced cox-2 gene expression were studied in a human keratinocyte cell line, HaCaT. UVB significantly increased cox 2 gene expression at both protein and mRNA levels. As we reported previously, p38 and ERK were significantly activated after UVB irradiation in HaCaT cells. In addition, treating the cells with p38 inhibitor SB202190 or MEK inhibitor PD98059 specifically inhibited UVB induced p38 or ERK activation, respectively. In this study, we further examined the roles of p38 and ERK in UVB induced cox-2 gene expression in HaCaT cells. We found that SB202190 strongly inhibited UVB induced COX-2 protein expression at different time points and various UVB doses. Furthermore, SB202190 markedly inhibited UVB induced cox-2 mRNA. Our data indicated that ERK did not play a role in UVB induced cox-2 gene expression in human keratinocytes since suppression of ERK did not significantly alter UVB induced increase of COX-2 protein and mRNA. These results suggested, for the first time, that activation of p38 is required for UVB induced cox-2 gene expression in human keratinocytes. Since cox-2 expression plays an important role in UV carcinogenesis, p38 could be a potential molecular target for chemoprevention of skin cancer. PMID- 11439358 TI - Neuropathology of paediatric chronic intestinal pseudo-obstruction and related animal models. AB - Chronic intestinal pseudo-obstruction (CIP) in paediatric patients is due to heterogeneous aetiologies that include primary disorders of the enteric nervous system. These conditions are poorly delineated by contemporary diagnostic approaches, in part because the complex nature of the enteric nervous system may shelter significant physiological defects behind subtle or quantitative anatomical changes. Until recently, relatively few experimental animal models existed for paediatric CIP. However, the availability of rodent models, particularly novel mutants created in the last few years by genetic manipulations, has brought unprecedented opportunities to investigate molecular, cellular, physiological, and histological details of enteric neuropathology. Information gleaned from studies of these animals is likely to change diagnostic and therapeutic approaches to paediatric CIP and related conditions. PMID- 11439359 TI - Molecular markers of metastasis in squamous carcinomas. AB - The prognosis for patients with squamous carcinomas of the head and neck region is strongly influenced by the presence or absence of cervical lymph node metastases. Biological markers that indicated a high risk of metastasis would have a major role in determining the optimal therapeutic strategy for these patients. This editorial discusses recent studies of molecular and genetic markers of squamous carcinomas in relation to metastatic potential and highlights some of the issues that limit current progress in this field of study. PMID- 11439360 TI - Histological grading of oral epithelial dysplasia: revisited. AB - Treatment of oral precancer is largely based on histological grading of epithelial dysplasia, despite the fact that this estimation is subjective and therefore carries a low reproducibility. The grade of epithelial dysplasia may not be proportional to the risk of malignant potential and clinical characteristics may complement therapeutic decisions. Molecular genetic changes found in oral epithelial dysplasia remain unclear and at present lack clinical significance. Genomic and proliferation markers are likely to be associated with histopathological parameters, but their relationship with the grading of dysplasia remains uncertain. They are potential biomarker candidates but their utility in prognosis of oral precancer deserves further study. PMID- 11439361 TI - Expression of genetic markers in lymph node metastases compared with their primary tumours in head and neck cancer. AB - Regional metastasis is an important factor in the prognosis and treatment of head and neck squamous cell carcinoma (HNSCC). The results of earlier studies suggested the possibility of predicting nodal metastasis in HNSCC using biological markers. To identify which factors may be relevant in the metastatic behaviour of these tumours, the expression of several markers involved in tumour progression was studied in both nodal metastases and their corresponding primary tumours. Expression of p53, Rb, cyclin D1, myc, bcl-2, EGFR, neu, E-cadherin, epithelial cell adhesion molecule (Ep-CAM), and nm23 was studied in 54 primary tumours and their corresponding metastases in patients with HNSCC. The expression of most genes involved in tumourigenesis (p53, Rb, cyclin D1, myc, bcl-2, EGFR, neu, and E-cadherin) was similar in primary tumours and metastases. The expression of nm23 and Ep-CAM was found to be more frequently lower than higher in metastases, compared with their primary tumours. Whereas most genetic alterations of primary tumours remain unchanged in metastases, expression of the cell adhesion molecule Ep-CAM and of nm23 is more frequently reduced than increased in metastases, compared with their primary tumours, suggesting relevance to the process of metastasis. This also implies differences in the regulation of markers involved in tumourigenesis and the process of metastasis. PMID- 11439362 TI - Comparison of histological grading and large-scale genomic status (DNA ploidy) as prognostic tools in oral dysplasia. AB - Approximately one in ten oral white patches (leukoplakia) are histologically classified as dysplasia, with a well-documented potential for developing into oral squamous cell carcinoma (OSCC). Histological grading in oral dysplasia has limited prognostic value, whereas large-scale genomic status (DNA ploidy, nuclear DNA content) is an early marker of malignant transformation in several tissues. Biopsies from 196 patients with oral leukoplakia histologically typed as dysplasia were investigated. Inter-observer agreement among four experienced pathologists performing a simplified grading was assessed by Cohen's kappa values. For 150 of the 196 cases, it was also possible to assess large-scale genomic status and compare its prognostic impact with that of histological grading. Disease-free survival was estimated by life-table methods, with a mean follow-up time of 103 months (range 4-165 months). The primary considered end point was the subsequent occurrence of OSCC. For grading of the total of 196 cases, kappa values ranged from 0.17 to 0.33 when three grading groups (mild, moderate, and severe dysplasia) were considered, and from 0.21 to 0.32 when two groups (low grade and high grade) were considered (p=0.41). For the 150 cases in which large-scale genomic status was also assessed, kappa values for the histological grading ranged from 0.21 to 0.33 for three grading groups and from 0.27 to 0.34 for two grading groups (p=0.47). In survival analysis, histological grading was without significant prognostic value for any of the four observers (p 0.14-0.44), in contrast to DNA ploidy (p=0.001). It is concluded that DNA ploidy in oral dysplasia has a practical prognostic value, unlike histological grading of the same lesions. PMID- 11439363 TI - Detailed gene expression analysis but not microsatellite marker analysis of 9p21 reveals differential defects in the INK4a gene locus in the majority of head and neck cancers. AB - The INK4a gene locus on chromosome 9p21 encodes two proteins, p16(INK4a) and p14(ARF), which influence cell cycle control regulated by pRb and p53. The objective of this study was to use different methods for the analysis of the incidence of changes at the INK4a locus in head and neck cancer (HNSCC). Primary tumours were analysed for allelic imbalances (AI) with microsatellite markers for chromosome 9, by immunohistochemistry (IHC) and IHC with enhanced sensitivity by tyramide signal amplification (TSA-IHC), and by RT-PCR. No homozygous deletions at 9p21 were detected. AI at 9p21, which was found in approximately 60% of the tumours, completely failed to indicate the functional inactivation of the two INK4a gene products. Immunostaining of normal squamous epithelia revealed very low levels of p16(INK4a), whereas p14(ARF) was readily detectable. In 160 tumours, IHC suggested a loss of p16(INK4a) expression in 90%. However, by TSA IHC, only 53.7% showed loss of p16(INK4a) expression, and this was consistent with the RT-PCR analyses. In 100 tumours analysed for both proteins, selective loss of p16(INK4a) occurred in 37%; loss of p14(ARF) was found in only 15%, and selective loss in only 4%; 11% of the tumours had lost both proteins. We conclude that only IHC with high sensitivity and the combined expression analysis of mRNAs and proteins is suitable for studying the role of INK4a in HNSCC. The INK4a gene expression defects are frequent but not universal and primarily affect p16(INK4a). Their clinical impact is still not clear. PMID- 11439364 TI - Detailed deletion mapping in sporadic breast cancer at chromosomal region 17p13 distal to the TP53 gene: association with clinicopathological parameters. AB - Chromosome 17p is among the most frequently deleted regions in a variety of human malignancies including breast cancer. This study has further refined the localization of a putative tumour suppressor gene (TSG) at 17p13 distal to the TP53 gene in breast carcinomas. It was found that 73% (37 of 51) of the breast tumours exhibited loss of heterozygosity (LOH) at one or more loci at 17p13. The allelic loss patterns of these tumours suggest the presence of at least seven commonly deleted regions on 17p13. The three most frequently deleted regions were mapped at chromosomal location 17p13.3-17p13.2 between the markers D17S831 and D17S1845 (56% LOH), at 17p13.1 between D17S1810 and D17S1832 (53% LOH), and at 17p13.1 between D17S938 and TP53 (55% LOH). A significant correlation was found between loss at 17p13 and tumour grade, size, proliferative activity, and oestrogen receptor (ER) status. Losses at 17p13 were seen more frequently in large and poorly differentiated tumours with high proliferative activity. These data support and extend previous reports on the presence of a putative TSG(s) at chromosomal region 17p13 distal to the TP53 gene and show that different subsets of LOH are associated with more aggressive tumour behaviour. PMID- 11439365 TI - Similarity in expression of cell cycle proteins between in situ and invasive ductal breast lesions of same differentiation grade. AB - There is increasing evidence that there are different progression routes leading to invasive breast cancer, depending on histology and differentiation grade. The aim of this study was to determine alterations in the expression of proteins involved in proliferation and apoptosis in non-invasive and invasive ductal breast lesions. Immunohistochemistry was performed on 106 usual ductal hyperplasias (UDH), 61 DCIS lesions and 53 invasive ductal breast carcinomas. Increased proliferation (Ki67), overexpression of cyclin D1, HER-2/neu, p21 and p53, and decreased expression of bcl-2 and p27 could already be found in UDH. Significant differences between UDH and DCIS lesions were found for only one protein when UDH was compared with well-differentiated DCIS (p27), for three proteins when compared with intermediately differentiated DCIS (p21, cyclin D1, Ki-67), and for all proteins when compared with poorly-differentiated DCIS. Comparing DCIS with invasive lesions of same differentiation grade, proliferation was elevated in the invasive lesions. Altered expression of the other proteins was in general only slightly increased in the invasive lesion compared with DCIS. The number of proteins with altered expression per lesion was highest in poorly differentiated lesions and was comparable between DCIS and invasive cancer of the same differentiation grade. In conclusion, the biggest changes in expression of these proliferation and apoptosis related proteins appear to occur during the transition from hyperplasia to DCIS; they probably play a minor role in the transition from DCIS to invasive breast lesion of same differentiation grade. Well-differentiated in situ and invasive breast lesions share many of the aberrations in expression of these proteins, as do poorly-differentiated in situ and invasive lesions. However, there are many differences between the well and poorly-differentiated lesions. This further supports the existence of different progression routes leading to breast cancer. PMID- 11439366 TI - Microsatellite alterations and target gene mutations in the early stages of multiple gastric cancer. AB - Multiple gastric cancers may develop through the same genetic background: the mutator pathway due to defects in DNA mismatch repair genes, or the suppressor pathway due to defects in tumour suppressor genes. To clarify the critical genetic events in the early stages of multiple gastric cancer development, 29 early and four advanced gastric cancers were examined from 12 patients. Microsatellite alterations were studied involving microsatellite instability (MSI) and loss of heterozygosity (LOH) at tumour suppressor loci, representative of the mutator pathway and the suppressor pathway, respectively, as well as mutations of target genes (TGF-beta RII, BAX, hMSH3, and E2F-4). MSI was determined in ten cancers (10/33; 30.3%) from seven patients (7/12; 58.3%). LOH was detected in six cancers (6/33; 18.2%) from five patients (5/12; 41.7%), most frequently at TP53, in four cancers (4/33; 12.1%) from four patients (4/12; 33.3%). In cases with multiple gastric cancers in the same stomach, the MSI status was generally the same, but in two patients (2/12; 16.8%) a tumour with MSI-H and another with LOH were found to co-exist in the same stomach. As for mutations of the target genes, it was found that E2F-4 was mutated in six cancers (6/33; 18.2%) from four patients (4/12; 33.3%). Furthermore, identical E2F-4 mutations were detected in four of the six intestinal metaplastic mucosae adjacent to each cancer carrying an E2F-4 mutation. No mutations were detected in the other target genes. In conclusion, the present results indicate that the majority of multiple gastric cancers develop from the same genetic background, with the mutator pathway playing a more important role than the suppressor pathway. Mutations of E2F-4 are early events in multiple gastric cancer development, occurring even in the intestinal metaplastic mucosa, with mutations of other target genes to follow during cancer progression. PMID- 11439367 TI - PTEN expression is maintained in sporadic colorectal tumours. AB - Loss of PTEN (phosphatase and tensin homologue deleted from chromosome 10) function has been implicated in the progression of several types of cancer. Allele loss close to the PTEN locus occurs in sporadic colon cancer and germline PTEN mutations cause Cowden disease, an inherited cancer syndrome characterized by an increased incidence of gastrointestinal tract lesions that can progress to colorectal carcinoma. However, although PTEN is a good candidate for involvement in the pathogenesis of sporadic colon cancer, previous analyses have not revealed a high frequency of somatic mutations in colorectal tumours. Alternative mechanisms which could lead to a loss of PTEN expression in colon cancer have not been investigated. This study monitored PTEN mRNA and protein levels in a panel of 50 tumour tissues obtained from 35 patients with sporadic colon cancer. RT-PCR and immunohistochemistry were used to evaluate the expression of mRNA and protein, respectively, in normal, adenoma and adenocarcinoma colorectal tissues as well as in metastatic lesions. To overcome the problem of heterogeneity and normal stromal cell contamination in homogenized tissue specimens, specific cell types were isolated by microdissection prior to PCR analysis. No loss of PTEN expression was evident in any of the colon tissues examined. PTEN protein was localized exclusively in the cytoplasm of normal and tumour cells and no correlation of immunostaining intensity and tumour stage or grade was revealed. As with previous deletion and mutation analyses, the present study suggests that loss of PTEN expression is not prevalent in sporadic colon cancer. PMID- 11439368 TI - Microarrays of bladder cancer tissue are highly representative of proliferation index and histological grade. AB - The number of genes suggested to play a role in cancer biology is rapidly increasing. To be able to test a large number of molecular parameters in sufficiently large series of primary tumours, a tissue microarray (TMA) approach has been developed where samples from up to 1000 tumours can be simultaneously analysed on one glass slide. Because of the small size of the individual arrayed tissue samples (diameter 0.6 mm), the question arises of whether these specimens are representative of their donor tumours. To investigate how representative are the results obtained on TMAs, a set of 2317 bladder tumours that had been previously analysed for histological grade and Ki67 labelling index (LI) was used to construct four replica TMAs from different areas of each tumour. Clinical follow-up information was available from 1092 patients. The histological grade and the Ki67 LI were determined for every arrayed tumour sample (4x2317 analyses each). Despite discrepancies in individual cases, the grade and Ki67 information obtained on minute arrayed samples were highly similar to the data obtained on large sections (p<0.0001). Most importantly, every individual association between grade or Ki67 LI and tumour stage or prognosis (recurrence, progression, tumour specific survival) that was observed in large section analysis could be fully reproduced on all four replica TMAs. These results show that intra-tumour heterogeneity does not significantly affect the ability to detect clinico pathological correlations on TMAs, probably because of the large number of tumours that can be included in TMA studies. TMAs are a powerful tool for rapid identification of the biological or clinical significance of molecular alterations in bladder cancer and other tumour types. PMID- 11439369 TI - Abnormalities of the E-cadherin/catenin adhesion complex in classical papillary thyroid carcinoma and in its diffuse sclerosing variant. AB - The cadherin/catenin complex regulates cellular adhesion and motility and is believed to function as an invasion suppressor system. Several studies have identified alterations in the expression profiles of those molecules in different histotypes of thyroid carcinoma. The diffuse sclerosing variant (DSV) of papillary thyroid carcinoma (PTC) is a rare, highly invasive variant of PTC in which an impairment of cell-cell adhesion may play a major role. In an attempt to progress in the understanding of the clinicopathological features of DSV, this study examined eight cases of DSV, 18 cases of classical PTC and a control group of normal thyroid by immunohistochemistry (E-, P- and N-cadherins and beta-, gamma- and alpha-catenins). The E-cadherin gene was also studied by polymerase chain reaction/single strand conformation polymorphism (PCR/SSCP) and methylation specific PCR (MSP). In contrast to classical PTC, which showed heterogeneous loss of E-cadherin expression, in almost every case of DSV a pronounced reduction was observed in its membranous expression, accompanied by a relocation to the cytoplasm. Inactivation of the E-cadherin/catenin complex appears to occur in DSV via two different pathways: E-cadherin alteration either through mutation (one out of the eight cases) or through methylation of the E-cadherin gene promoter (three out of five cases); and beta- and/or gamma-catenin alterations (three of the eight cases). Methylation of the E-cadherin gene promoter, abnormalities of E cadherin expression and alterations of gamma-catenin were also detected in classical PTC. In DSV, as in classical PTC, there is neo-expression of P-cadherin in areas of squamous metaplasia and no N-cadherin expression. In conclusion, abnormalities of the E-cadherin/catenin complex appear to be more pronounced in DSV than in classical PTC. It remains to be shown whether or not such differences are associated with the more aggressive behaviour of DSV compared with classical PTC. PMID- 11439370 TI - NF2 status of meningiomas is associated with tumour localization and histology. AB - In approximately 60% of sporadic meningiomas, the tumour suppressor gene NF2, located on chromosome 22q, is inactivated. Mutations in the NF2 gene have been specifically reported in transitional and fibrous, but not meningothelial, meningiomas. Since meningothelial meningiomas frequently occur in anterior parts of the skull base, the association between tumour localization, size, histological subtype and NF2 status was investigated in a group of 42 sporadic meningiomas. NF2 status was determined by LOH analysis, karyotyping and FISH. Tumour size and site were evaluated by CT scans and MRIs. A strong correlation between tumour localization in the anterior skull base and intact 22q was revealed (p=0.003). On the other hand, tumour localization at the convexity was associated with disruption of NF2 (p=0.023). Furthermore, an association between chromosome 22 status and histological subtype was observed: abnormalities of chromosome 22q were more frequent in transitional and fibrous meningiomas than in the meningothelial variant (p<0.001). Also, the meningothelial meningiomas were more often located in the anterior skull base (p<0.006). Based on these findings, it is concluded that an alternative histogenesis and genetic pathway is likely to exist for meningiomas arising in the anterior skull base. PMID- 11439371 TI - Human MUC1 mucin: a potent glandular morphogen. AB - Human MUC1 mucin is a high-molecular-weight transmembrane glycoprotein, which is apically expressed in the majority of glandular epithelia. During embryonic development, changes in the pattern of MUC1 mucin expression coincide with the onset of glandular differentiation. This mucin is also frequently overexpressed and aberrantly glycosylated in carcinomas. To investigate the potential role of MUC1 mucin in morphogenesis, a full length MUC1 cDNA was transfected into murine mammary adenocarcinoma (410.4) and Madin-Darby canine kidney (MDCK) cells. This generated four clonal cell lines. Western blotting, FACS analysis, and immunohistochemistry confirmed expression of MUC1. All four MUC1-expressing clones demonstrated altered morphogenesis when cultured in three-dimensional type I collagen gels. While parental and vector control 410.4 cells formed compact spherical structures, the MUC1-expressing clones formed complex branching structures. Similarly, while parental and vector control MDCK cells formed small circumscribed colonies with a central lumen, the MUC1-expressing clones formed elongated tubules. MUC1 expression was also associated with reduced cellular cohesion and enhanced migration on type I collagen-coated surfaces for all except one of the clones, which expressed only low levels of MUC1 on the cell surface. These results show that MUC1 expression stimulates morphogenetic changes in two distinct epithelial cell lines. Taken together with previous observations on MUC1 expression in embryonic development and carcinomas, this finding suggests that MUC1 may induce changes in tissue architecture in both normal development and cancer. PMID- 11439372 TI - Hyaluronan synthases, hyaluronan, and its CD44 receptor in tissue around loosened total hip prostheses. AB - Aseptic loosening of prosthetic components, the most common long-term complication after total hip replacement (THR), is characterized by the formation of a synovial membrane-like interface tissue (SMLIT). It was hypothesized that the hyaluronan synthase (HAS)/hyaluronan (HA)/HA receptor CD44 signalling system is responsible for the synovial-like differentiation of the interface membrane. SMLIT was therefore compared with osteoarthritis (OA) synovial membrane by using reverse transcriptase polymerase chain reaction (RT-PCR) of HAS 1, 2 and 3, histochemical HA assay, and immunohistochemistry of CD44 and its non-HA ligands. All three isoforms of HAS were found in these samples. HA and CD44 were most abundant in the lining, but the signal was actually stronger in aseptic loosening than in OA (p<0.01). The non-HA CD44 ligands, collagen type VI, fibronectin, osteopontin, and MCP-1, had a similar distribution pattern in both tissues. These results confirm the synovial-like structure of the interface tissue lining. The pressure waves and movement of the HA-rich pseudosynovial fluid seem to drive HA into the implant-to-host interface, which itself also produces HA. HA may be responsible for the induction of a synovial-like lining at the interface through HA-CD44 signalling. PMID- 11439373 TI - Why do we faint? PMID- 11439374 TI - Muscle glycogenoses. AB - There are 11 hereditary disorders of glycogen metabolism affecting muscle alone or together with other tissues, and they cause two main clinical syndromes: episodic, recurrent exercise intolerance with cramps, myalgia, and myoglobinuria; or fixed, often progressive weakness. Great strides have been made in our understanding of the molecular bases of these disorders, all of which show remarkable genetic heterogeneity. In contrast, the pathophysiological mechanisms underlying acute muscle breakdown and chronic weakness remain unclear. Although glycogen storage diseases have been studied for decades, new biochemical defects are still being discovered, especially in the glycolytic pathway. In addition, the pathogenesis of polyglucosan deposition is being clarified both in traditional glycogenoses and in disorders such as Lafora's disease. In some conditions, combined dietary and exercise regimens may be of help, and gene therapy, including recombinant enzyme replacement, is being actively pursued. PMID- 11439375 TI - Role of adaptive plasticity in recovery of function after damage to motor cortex. AB - Based upon neurophysiologic, neuroanatomic, and neuroimaging studies conducted over the past two decades, the cerebral cortex can now be viewed as functionally and structurally dynamic. More specifically, the functional topography of the motor cortex (commonly called the motor homunculus or motor map), can be modified by a variety of experimental manipulations, including peripheral or central injury, electrical stimulation, pharmacologic treatment, and behavioral experience. The specific types of behavioral experiences that induce long-term plasticity in motor maps appear to be limited to those that entail the development of new motor skills. Moreover, recent evidence demonstrates that functional alterations in motor cortex organization are accompanied by changes in dendritic and synaptic structure, as well as alterations in the regulation of cortical neurotransmitter systems. These findings have strong clinical relevance as it has recently been shown that after injury to the motor cortex, as might occur in stroke, post-injury behavioral experience may play an adaptive role in modifying the functional organization of the remaining, intact cortical tissue. PMID- 11439376 TI - An innovative hand brace for carpal tunnel syndrome: a randomized controlled trial. AB - We developed a hand brace and studied its efficacy and tolerability in patients with carpal tunnel syndrome (CTS). We randomized 83 subjects into a treated group, which wore the hand brace at night for 4 weeks, and a control group, which received no treatment. The primary efficacy measure was change in the Boston Carpal Tunnel Questionnaire (BCTQ) score. Secondary measures were Subjects' Global Impression of Change Questionnaire (SGICQ), median distal motor latency, sensory conduction velocity and amplitude, and neurophysiological class of severity. The treated group showed a reduction in BCTQ symptomatic score (from 2.75 to 1.54 at 4 weeks; P < 0.001) and functional score (from 1.89 to 1.48; P < 0.001). There were no significant changes in the control subjects. SGICQ documented improvement in all treated subjects (P = 0.006). No significant difference was found in electrophysiological measurements, but overall neurophysiological classification shifted to less severe classes in the treated group (P < 0.05). Thus, the study demonstrates that this hand brace is highly efficient in relieving symptoms and functional loss in CTS. PMID- 11439377 TI - Sodium influx during action potential in innervated and denervated rat skeletal muscles. AB - Resting Na(+) influx (J(i)(Na)) was measured in innervated and denervated (1-6 days) rat extensor digitorum longus muscle in the absence and presence of 2 micromol/L tetrodotoxin (TTX). The mean value of Na(+) permeability (P(Na)) in innervated muscles was 49.6 +/- 2.6 pm.s(-1). At the second day postdenervation, it decreased by about 45%. This was followed, between the second and fourth days, by a sharp rise, which by the sixth day reached a steady value approximately 2.5 times greater than that of innervated muscles. This, most likely, generated the 30% increase in internal [Na(+)] concentration ([Na(+)](I)) observed at this time. Tetrodotoxin reduced P(Na) of both innervated and denervated muscles by about 25%. In 6-day denervated muscles, virtually all the TTX effect on P(Na) represents the blockage of TTX-resistant Na(+) channels. Denervation produced a depolarization of about 20 mV by the sixth day. The extra J(i)(Na) per action potential (AP) decreased monotonically with time after denervation from 20.0 +/- 3.8 in innervated to 11.1 +/- 1.0 nmol.g(-1).AP(-1) in 6-day denervated muscles. The overshoot of the AP decreased from 15 +/- 1 in innervated to 7 +/- 1 mV in 6 day denervated muscles. Likewise, the maximum rate of rise (+dV/dt), an expression of the inward Na(+) current, fell from 305 +/- 14 in innervated to 188 +/- 18 V.s(-1) in 6-day denervated muscles. The estimated 6-day denervated/innervated ratio of peak Na(+) conductance (g(Na)) was 0.67. The changes in AP parameters promoted by denervation were substantially reduced when both innervated and denervated fibers were hyperpolarized to -90 mV. These results suggest that the depolarization, mainly due to the increase in P(Na) /P(K) ratio, increases Na(+) inactivation and consequently reduces peak g(Na), in spite of the absolute increment in resting TTX-sensitive P(Na). This, in addition to the moderate reduction in the inward driving force on Na(+), decreases the inward Na(+) current and the extra J(i)(Na) per AP. PMID- 11439378 TI - Epidermal nerve fiber density in sensory ganglionopathies: clinical and neurophysiologic correlations. AB - We assessed the involvement of somatic unmyelinated fibers in sensory ganglionopathies by skin biopsy and quantitative sensory testing (QST). Sixteen patients with ganglionopathy, 16 with axonal neuropathy, and 15 normal controls underwent skin biopsy at the proximal thigh and the distal leg. Intraepidermal nerve fibers (IENF) were immunostained by antiprotein gene product 9.5, and their linear density was quantified under light microscopy. Confocal microscopy studies with double staining of nerve fibers and basement membrane were also performed. Healthy subjects and neuropathy patients showed the typical proximodistal gradient of IENF density; in neuropathies, values were significantly lower at the distal site of the leg, confirming the length-dependent loss of cutaneous innervation. Conversely, ganglionopathy patients with hyperalgesic symptoms did not show any change of IENF density between the proximal thigh and the distal leg. The distinct pattern of epidermal denervation seen in sensory ganglionopathy reflected the degeneration of somatic unmyelinated fibers in a fashion that was not length-dependent, which was consistent with both clinical and neurophysiologic observations and supported the diagnosis. PMID- 11439379 TI - Quadriceps muscle function and fatigue in women with Addison's disease. AB - In nine patients with Addison's disease (mean +/- SE: 51 +/- 2 years) receiving conventional steroid treatment, and nine age-matched healthy controls (56 +/- 2 years), we investigated maximum voluntary quadriceps force (MVC) and contractile properties evoked with stimulation and central activation both at rest and during a submaximal intermittent fatigue task. The MVC was similar (-3%), but twitch tension (-27%) and central activation were significantly less (-7%), and tetanic half-relaxation time was approximately 40% slower in the patients. Twitch amplitudes were potentiated by 6% in the patients, but unchanged in the control group. The patients self-terminated a submaximal intermittent fatigue protocol (0.6 duty cycle) at approximately 5 +/- 1 min, whereas the controls stopped when they lost 50% of MVC force ( approximately 10 +/- 1 min). Force loss was similar between groups over the first 5 min of the fatigue task. In the patient group, maximal and submaximal relative integrated electromyogram (IEMG) increased significantly in the first minute of fatigue and remained elevated, whereas the controls exhibited a gradual increase in submaximal IEMG with little change in maximal IEMG. These results indicate that conventionally treated Addison's patients have similar MVC strength, but altered contractile properties and decreased endurance compared with controls. PMID- 11439380 TI - Thalidomide neuropathy in patients treated for metastatic prostate cancer. AB - We prospectively evaluated thalidomide-induced neuropathy using electrodiagnostic studies. Sixty-seven men with metastatic androgen-independent prostate cancer in an open-label trial of oral thalidomide underwent neurologic examinations and nerve conduction studies (NCS) prior to and at 3-month intervals during treatment. NCS included recording of sensory nerve action potentials (SNAPs) from median, radial, ulnar, and sural nerves. SNAP amplitudes for each nerve were expressed as the percentage of its baseline, and the mean of the four was termed the SNAP index. A 40% decline in the SNAP index was considered clinically significant. Thalidomide was discontinued in 55 patients for lack of therapeutic response. Of 67 patients initially enrolled, 24 remained on thalidomide for 3 months, 8 remained at 6 months, and 3 remained at 9 months. Six patients developed neuropathy. Clinical symptoms and a decline in the SNAP index occurred concurrently. Older age and cumulative dose were possible contributing factors. Neuropathy may thus be a common complication of thalidomide in older patients. The SNAP index can be used to monitor peripheral neuropathy, but not for early detection. PMID- 11439381 TI - Regenerating soleus and extensor digitorum longus muscles of the rat show elevated levels of TNF-alpha and its receptors, TNFR-60 and TNFR-80. AB - We measured the mRNA and protein levels of tumor necrosis factor-alpha (TNF alpha) and the transcript levels of its receptors (TNFR-60 and TNFR-80) in the rat soleus (slow twitch) and extensor digitorum longus (EDL; fast twitch) muscles regenerating from notexin-induced necrosis. On the first day after administration of the toxin, when most fibers were necrotic and invaded by inflammatory cells/macrophages, dramatic increases of transcript and protein levels of TNF alpha and of the mRNA levels of its receptors were observed. The transcript levels of TNF-alpha and TNFR-60, but not of TNFR-80, showed a second but smaller increase at the time when newly formed muscle fibers became reinnervated. In situ hybridization showed that on day 1, during the phase of extensive necrosis, the transcript of TNF-alpha was abundantly present and on day 4 of regeneration it was most often seen in areas devoid of desmin. The mRNA level of TNF-alpha was not detectable in BC(3)H1- and C2C12-cultured myoblasts and it was low in freeze injured muscle, corresponding to the relatively mild degree of inflammation elicited by freezing. Therefore, our results are most consistent with the view that inflammatory cells/macrophages are the main source of TNF-alpha. PMID- 11439382 TI - Brachial plexus neoplastic lesions assessed by conduction study of medial antebrachial cutaneous nerve. AB - Two cases of neoplastic involvement of the lower brachial plexus are reported. This condition was due to recurrence of lymphoma in one case and to axillary node spread of breast cancer in the other. The neuropathic origin and the location of the lesion in the lower brachial plexus between the T-1 root and the axilla was demonstrated by the presence of abnormalities on testing of the medial antebrachial cutaneous nerve in the symptomatic upper limb and comparing it to the healthy one. All other electrodiagnostic tests were normal. Such a finding suggests the need for imaging of the lower brachial plexus region by computed tomography or magnetic resonance imaging. PMID- 11439383 TI - Maximal exercise and muscle energy metabolism after recovery from exercise hyperthermia syndrome. AB - Muscle energy metabolism was studied in 30 subjects after recovery from exercise hyperthermia syndrome (EHTS subjects) and 15 healthy men with identical physical activities. Blood lactate, free fatty acid (FFA), serum creatine kinase activity (CK), and glycerol and the temperature in the auditory duct (T(c)) and on the thumb pad (T(sk)) were measured at rest and during and after maximal exercise on a cycloergometer. The EHTS subjects had a limitation of physical performance, with lowered values for maximal oxygen uptake (VO(2max), P < 0.0005), maximal workload (P < 0.05), and ventilatory threshold (V(t), P < 0.0005). The discrepancy between high plasma concentrations of FFA and the lack of decrease in respiratory ratio (RR) suggests that, in EHTS subjects, a very active release of FFA was not balanced by a proportional increase in catabolism. The increased skin temperature was smaller in EHTS subjects (P < 0.05 at 180 and 200 W). At the end of exercise, auditory duct temperature increase was higher in EHTS subjects than in control subjects (P < 0.05). This study thus showed an impairment of muscle metabolism and an abnormality of thermoregulatory mechanisms. These results may provide insight into the underlying physiopathological disturbance. PMID- 11439384 TI - When to use the combined sensory index. AB - A recently developed electrodiagnostic technique, the combined sensory index (CSI), has been recommended for its greater sensitivity in diagnosing carpal tunnel syndrome (CTS). The CSI requires a greater number of procedures and therefore involves greater time, cost, and patient discomfort than does conventional electrodiagnostic testing. The CSI is composed of three commonly used electrodiagnostic techniques. There is a close correlation between the components of the CSI, and in most cases, all three components of the CSI are in agreement. We performed a study to develop and validate an algorithm that could be used to identify subsets of patients with CTS in whom CSI testing is particularly useful. Subjects were consecutive outpatient veterans referred by a heterogeneous group of specialists and generalists for electrodiagnostic evaluation of paresthesias in a median distribution with nocturnal exacerbation of symptoms. The CSI served as our gold standard. Using our simple algorithm, we found that in approximately 95% of cases, it was unnecessary to perform the CSI. This management strategy improves patient comfort and reduces electrodiagnostic cost while identifying the minority of patients for whom the CSI is indicated. PMID- 11439385 TI - Cell death and apoptosis-related proteins in muscle biopsies of sporadic amyotrophic lateral sclerosis and polyneuropathy. AB - To investigate disease-related differences of cell death and apoptosis in human denervation atrophy, we studied DNA fragmentation by the terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) method in 38 biopsies of clinically nonaffected and affected muscles from patients with sporadic amyotrophic lateral sclerosis (sALS), in 13 muscle biopsies from patients with chronic peripheral neuropathies, and in 8 biopsies from control subjects. In addition, expression of apoptosis-related proteins, bax, bcl-2, and Fas, was studied in 20 biopsies of sALS and 10 chronic peripheral neuropathies. We identified DNA cleavage in 10% of myofibers of patients and in up to 1.5% of control samples. In clinically affected muscles of ALS, a larger amount of TUNEL positive myofibers (mean 10.5 +/- 5.9%) was detected, similar to chronic peripheral neuropathies (mean 10.0 +/- 7.4%). Atrophic myofibers were immunopositive for bax, bcl-2, and, to a weaker extent, for Fas. However, bax-, bcl-2-, or Fas-positive atrophic myofibers did not reveal consecutive DNA cleavage. Differences between sALS subgroups and chronic peripheral neuropathies were not found. In human denervation atrophy the bcl-2/bax and the FasL/Fas systems are apparently active independently of DNA fragmentation and apoptosis. DNA fragmentation thus displays an additional reaction that is not disease specific at chronic stages of human denervation processes, probably recapitulating events like skeletal muscle fiber remodeling in embryonic skeletal tissue development. PMID- 11439386 TI - Passive stiffness is increased in soleus muscle of desmin knockout mouse. AB - The effects of an absence of desmin on passive stiffness of skeletal muscle were analyzed using soleus muscles from desmin knockout mice. Stiffness was evaluated by analyzing the passive tension induced by a ramp-and-hold extension test. This test showed that passive resistance to stretch was largely increased for muscles lacking desmin. This study could facilitate interpretation of changes in muscle mechanics observed in humans affected by desmin-related myopathies. PMID- 11439387 TI - Brachial plexus involvement as the only expression of hereditary neuropathy with liability to pressure palsies. AB - Hereditary neuropathy with liability to pressure palsies (HNPP) is an autosomal dominant focal neuropathy characterized by recurrent entrapment neuropathies. Single members of families with HNPP and brachial plexus involvement have been reported previously. We describe a family with three affected members and in which the only symptom of entrapment was recurrent brachial plexopathy. The diagnosis of HNPP in this family was confirmed by both neurophysiological methods and DNA analysis. The distinction between HNPP and hereditary neuralgic amyotrophy is discussed. PMID- 11439388 TI - Test-retest reliability of combined sensory index. PMID- 11439392 TI - Occupation and leukemia: a population-based case-control study in Iowa and Minnesota. AB - BACKGROUND: Studies have suggested that risk of leukemia may be associated with occupational or industrial exposures and risk may vary by the histological type of the disease. METHODS: A population-based case-control study was conducted in Iowa and Minnesota to evaluate the association between various occupations, industries, and occupational exposures and leukemia risk. A total of 513 cases and 1,087 controls was included in the study. A lifetime occupational history and other risk factor information were collected through in-person interviews, and a job-exposure matrix was used to assess possible risks associated with specific exposures. RESULTS: A significantly increased risk of leukemia was observed among agricultural service industries and among nursing and healthcare workers. Janitors, cleaners, and light truck drivers also experienced increased risk. Those employed in plumbing, heating and air conditioning industries, and sales of nondurable goods (such as paints and varnishes) had an increased risk. Printers, painters, and workers in the food and metal industries had a nonsignificantly increased risk of leukemia. Analyses by specific exposures and histology of leukemia showed that risk of leukemia associated with occupational or industrial exposures may vary by histological type of the disease. CONCLUSIONS: An increased risk of leukemia among workers employed in agricultural industries, nursing and healthcare workers, and in a few occupations with possible exposure to solvents is consistent with earlier studies. Associations of risk with occupations not observed previously deserve further assessment. Published 2001 Wiley-Liss, Inc. PMID- 11439393 TI - Assignment of work involving farm tractors to children on North American farms. AB - BACKGROUND: Children are at high risk for tractor-related injury. The North American Guidelines for Children's Agricultural Tasks (NAGCAT) provide recommendations for the assignment of tractor work. This analysis describes tractor-related jobs assigned to farm children and compares them to NAGCAT. METHODS: A descriptive analysis was conducted of baseline data collected by telephone interview during a randomized, controlled trial. RESULTS: The study population consisted of 1,138 children who worked on 498 North American farms. A total of 2,389 farm jobs were reported and 456 (19.1%) involved operation of farm tractors. Leading types of tractor jobs were identified. Modest, yet important, percentages of children were assigned tractor work before the minimum ages recommended by NAGCAT. CONCLUSIONS: Children on farms are involved in tractor work at a young age and some are involved in jobs that they are unlikely to have the developmental abilities to perform. NAGCAT is a new parental resource that can be applied to these work situations. PMID- 11439394 TI - Predictors of hearing loss in New York farmers. AB - BACKGROUND: Data from the telephone interview portion of the New York Farm Family Health and Hazard Surveillance were used to study self-reported hearing loss in New York farmers. METHODS: One thousand six hundred and twenty-two persons completed the hearing loss and noise exposure interview. Hearing loss was defined as at least some trouble hearing in one or both ears. Predictors of hearing loss were determined using logistic regression. RESULTS: Twenty-two percent of participants reported hearing loss. From the logistic regression, significant confounders are age (P = 0.0001), gender (P = 0.0001), being from a livestock farm (P = 0.012), and loss of consciousness due to head trauma (P = 0.04). Significant noise exposures are more hours of lifetime exposure to noisy farm equipment (P = 0.001) and having had a noisy non-farm job (with some hearing protection P = 0.002, without any hearing protection P = 0.0001). CONCLUSIONS: Farm noise exposure is a serious risk to the hearing of this population. Although use of hearing protection should be encouraged, replacing and modifying farm equipment to decrease noise at the source should be the first priority. PMID- 11439395 TI - Severe farm injuries among New York farmers. AB - BACKGROUND: Data from the telephone interview portion of the New York State Farm Family Health and Hazard Surveillance were used to study the incidence and predictors of severe farm injury. METHODS: One thousand seven hundred and six participants completed two telephone interviews in which they reported all injuries over a 12-month period. RESULTS: Nine percent of participants reported at least one severe farm injury. Using logistic regression the significant risk factors for sustaining at least one severe farm injury are younger age, the presence of hearing loss or joint trouble, working more hours per day, being the owner/operator of the farm, and being from a farm with higher gross sales. CONCLUSIONS: There needs to be continuing education of all farmers as to the risks of injury. However, when resources are limited, we recommend that injury education and interventions in this farming population should target younger farmers, those who work longer hours, owner/operators, farmers from higher grossing farms, with special attention to farmers who have physical impairments. PMID- 11439396 TI - Neuropsychological function in Gulf War veterans: relationships to self-reported toxicant exposures. AB - BACKGROUND: The present study was aimed at (1) exploring evidence of central nervous system (CNS) dysfunction among Gulf War (GW) veterans on neuropsychological tests and (2) examining whether performance on neuropsychological tests was related to specific neurotoxicant exposures experienced in the Gulf. METHODS: The GW-deployed groups were selected using stratified random sampling methods from two distinct cohorts of GW veterans. A comparison group that had been called up for GW service but deployed to Germany rather than the Gulf also was examined. Neuropsychological function was assessed using a pre-determined battery chosen to include tests known to be highly sensitive to the behavioral effects of the neurotoxicants thought to have been present in the Gulf. RESULTS: Self-reported exposures were related to neuropsychological test performance controlling for post-traumatic stress disorder, major depression, and other known covariates of neuropsychological test performance. Results showed that GW-deployed veterans performed more poorly than the Germany-deployed veterans on several specific neuropsychological tests, but after adjustment for multiple comparisons, only the differences in mood complaints remained significant. Within the GW-deployed group, self-reported exposure to chemical warfare agents was associated with poorer performance on cognitive tests involving specific functional domains. CONCLUSIONS: Results provide evidence that there are subtle differences in CNS function among GW deployed veterans who report chemical warfare agent exposure while in the GW theater. PMID- 11439397 TI - Respiratory function in active firefighters. AB - BACKGROUND: Exposure to environmental pollution during firefighting may results in the development of respiratory disorders in firefighters. METHODS: The health effects of firefighting on respiratory function was investigated in a group of 128 active firefighters by recording respiratory symptoms and measuring lung function. In addition, 88 control workers, not exposed to known pollutants were studied for the prevalence of acute and chronic respiratory symptoms. RESULTS: Significantly higher prevalences of dyspnea, nasal catarrh, sinusitis, and hoarseness were recorded in firefighters compared to control workers (P < 0.01). One subject developed asthma symptoms following two intense firefighting episodes. A high prevalence of acute symptoms experienced during and after fire extinguishing was also documented among these firefighters. Eye and throat irritation as well as headache were prominent. A logistic regression analysis of chronic respiratory symptoms demonstrated that odds ratios were significant for both duration of work exposure and for smoking. Lung function testing demonstrated a decrease in FEF75 in relation to predicted suggesting obstructive changes in the smaller airways. A regression analysis of ventilatory capacity tests indicated a positive relationship of forced vital capacity with length of employment, 1 s forced expiratory volume as well as FEF50 were related to smoking, and FEF75 was related to both smoking and length of employment. CONCLUSIONS: Our data suggest that firefighters are at risk for developing acute and chronic respiratory symptoms as well as obstructive airway changes. PMID- 11439398 TI - Blood pressure, left ventricular mass, and lead exposure in battery manufacturing workers. AB - BACKGROUND: Although debate about the relationship between lead and blood pressure has focused on low environmental lead levels, industrial exposure remains a concern. METHODS: We measured blood pressure and left ventricular mass (LVM) in 108 battery manufacturing workers, and calculated cumulative and historic average measures of blood lead. RESULTS: Diastolic pressure increased with increasing lead levels, with a significant (P = 0.04) 5 mmHg difference in mean pressure between the highest and lowest cumulative exposure levels. Diastolic pressure increased with the log of cumulative lead (P = 0.06). Both hypertension (defined as currently medicated or systolic > 160 mmHg or diastolic > 95 mmHg) and LVM increased nonsignificantly with increasing lead exposure (P values > or = 0.17 for hypertension and > or = 0.20 for LVM). CONCLUSIONS: We found a small effect of blood lead on diastolic blood pressure, particularly for a cumulative measure of exposure, but no convincing evidence of associations between lead and other blood-pressure-related outcomes. Published 2001 Wiley Liss, Inc. PMID- 11439399 TI - Respiratory symptoms and lung function in workers in heavy and highway construction: a cross-sectional study. AB - BACKGROUND: Occupational exposures for workers in heavy and highway (HH) construction include cement-containing dusts and diesel exhaust (DE). To investigate possible health effects, respiratory symptoms and lung function were examined in laborers, tunnel workers (TW), and operating engineers (OE) in HH and tunnel construction. The principal outcome of interest was airways disease. METHODS: Subjects were recruited through their unions. Medical and occupational histories and flow-volume loops were obtained. Based on self-report, asthma and chronic bronchitis were categorized as (1) physician-diagnosed or (2) for asthma, undiagnosed likely, and (3) for chronic bronchitis, symptomatic. Trade and time in the union were used as surrogates of exposure. Prevalence of asthma and chronic bronchitis, lung function outcome, and relationships with exposure variables were examined. RESULTS: Data were obtained on 389 workers: 186 laborers, 45 TWs, and 158 OEs. Prevalence of asthma was 13 and 11.4% for laborers (including TW) and OEs, respectively, and of symptomatic chronic bronchitis, 6.5 and 1.9%, respectively. Odds ratios (OR) for undiagnosed asthma likely were significantly elevated in TWs compared to OEs, and marginally elevated for chronic bronchitis. Inverse relationships were observed between time in the union, and risk for asthma and chronic bronchitis. Asthma (physician-diagnosed or undiagnosed likely) predicted lower FEV(1). Current cigarette use was associated with chronic bronchitis but not asthma. CONCLUSIONS: TWs, laborers, and OEs in HH construction are at increased risk for asthma. TWs also appear to be at increased risk for chronic bronchitis. Our data suggest that symptomatic workers are self selecting out of their trade. Asthma was associated with lower lung function in those affected. PMID- 11439400 TI - Accelerated silicosis in workers exposed to agate dust in Guangzhou, China. AB - BACKGROUND: An investigation of a small private agate mill was prompted by an agate worker who presented with silicosis in Guangzhou, China, in December 1998. METHODS: The work processes and records of dust measurements of the mill were examined. The mean total dust concentrations ranged from 3.0 to 9.9 mg/m3; 86-88% of the particles' diameter was smaller than 5.0 microm. Free SiO2 content from agate samples was measured. Occupational history was obtained and X-ray chest and lung function was carried out. RESULTS: Free SiO2 content of the agate was 90.5%. Thirty-two men involved in processing agate stone were examined. The mean ( +/- SD) age was 29.8 ( +/- 4.9) years and the mean (+/- SD) duration of exposure was 3.5 (+/- 1.7) years. Fifteen cases (47%) were diagnosed as accelerated silicosis. Up to September 1999, three had died from respiratory failure and five were in critical condition. CONCLUSIONS: Silicosis is an important problem in primitive work environments. PMID- 11439401 TI - Occupational cancer genetics: infrequent ras oncogenes point mutations in lung cancer samples from chromate workers. AB - BACKGROUND: Chromium carcinogenicity and mutagenicity are no longer disputed. However, although chromium has various genetic effects that induce cancer, its mechanism of inducing lung cancer in humans is still not fully understood. p53, a tumor suppressor gene, was found to be infrequently mutated in samples of lung cancer in workers with long occupational exposure to chromium, suggesting other cancer-related genes to be targeted in such tumors. METHODS: To assess the contribution of the ras oncogenes in the pathogenesis of chromate-related lung cancer, we studied point mutations at the critical positions of codons 12, 13, and 61 of the Ha-ras and Ki-ras oncogenes in 38 lung cancer samples derived from Japanese patients who worked in the chromate industry for long periods. We used both radioactive isotope and non-radioisotope PCR-SSCP techniques. RESULTS: The results of this study demonstrated that activation of ras genes due to point mutations in chromate-related lung cancer is a rare event. CONCLUSIONS: Ras oncogenes activated by point mutations do not have a major role in the process of tumorigenesis of chromate-related lung cancer. PMID- 11439402 TI - Availability and quality of industry and occupation information in the Massachusetts Cancer Registry. AB - BACKGROUND: Industry and occupation (I/O) data from state cancer registries can be useful in generating and evaluating hypotheses about occupational cancer risks. The quality of these data is uncertain. METHODS: We reviewed medical records on 1,020 cases from the Massachusetts Cancer Registry (MCR). We compared the presence and detail of I/O data from this dedicated record review (DRR) with routine reporting to the MCR in order to evaluate the potential to increase the availability of I/O data. RESULTS: Compared to routine reporting, the DRR increased the percentage of codable I/O data from 63.6 to 80.4%. The DRR also provided more detail in existing codes in 15.4% of cases. Other sections of the medical record in addition to the face sheet and sections of the record from the period before cancer diagnosis were important sources of additional information. Duration and timing of the corresponding I/O information were rarely recorded. CONCLUSIONS: The nearly one-third (32%) of records that had some improvement in I/O information with a dedicated review indicates considerable opportunity to improve the utility of this resource. PMID- 11439403 TI - Peripheral neuropathy in workers exposed to nitromethane. AB - BACKGROUND: Two workers from a headlight subassembly plant developed severe peripheral neuropathy. These workers had extensive, but brief (1-2 months) dermal and inhalational exposure to nitromethane, a solvent. METHODS: Environmental sampling was performed for nitromethane and ethyl cyanoacrylate. Medical records, including electrodiagnostic studies, were reviewed. Literature on nitromethane, ethyl cyanoacrylate, and other exposures in the workplace was reviewed. RESULTS: Electromyography and nerve conduction studies performed on these patients were consistent with a severe, axonal neuropathy. No etiology was discovered despite an extensive medical evaluation. Environmental sampling revealed exposure to nitromethane at the threshold limit value. CONCLUSIONS: The history of acute onset of severe peripheral neuropathy temporally associated with exposure to nitromethane is suggestive of a toxic neuropathy. While it cannot be definitively concluded that these two workers developed peripheral neuropathy secondary to exposures at work, occupational exposure to nitromethane appears to be the most likely etiology. PMID- 11439404 TI - Prognostic value of duration of illness and early intervention in bulimia nervosa: a systematic review of the outcome literature. AB - OBJECTIVE: Early intervention is often regarded as an important step for the prevention of a chronic eating disorder. The primary aim of this review was to systematically evaluate the outcome literature and to better describe the effects of early intervention on the course of bulimia nervosa. METHOD: Twenty-four follow-up studies were subjected to nine "best-evidence" criteria for methodological soundness. Of these 24 studies, 5 met all methodological criteria and were considered in more detail. RESULTS: Only one of these five studies found a significant association between duration of illness and outcome. These studies were re-examined to exclude studies that included patients who were not first admissions in order to distinguish between the effect of duration of illness versus early intervention. This re-examination excluded four of the five studies, and the remaining study provided no data regarding prior treatment. DISCUSSION: Based on this selection of studies, it was concluded that there is no consistent evidence to support that early intervention necessarily implies a better long term outcome. Due to the multiple methodological problems that plague follow-up studies, future research would be improved by employing designs that directly test the impact of early versus late intervention. PMID- 11439405 TI - Subtyping binge eating-disordered women along dieting and negative affect dimensions. AB - OBJECTIVE: Because etiologic and maintenance models of binge eating center around dieting and affect regulation, this study tested whether binge eating-disordered (BED) individuals could be subtyped along dieting and negative affect dimensions and whether subtypes differed in eating pathology, social functioning, psychiatric comorbidity, and response to treatment. METHOD: Three independent samples of interviewer-diagnosed BED women (N = 218) were subtyped along dieting and negative affect dimensions using cluster analysis and compared on the outcomes of interest. RESULTS: Cluster analyses replicated across the three independent samples and revealed a dietary subtype (63%) and a dietary-depressive subtype (37%). The latter subtype reported greater eating and weight obsessions, social maladjustment, higher lifetime rates of mood, anxiety, and personality disorders, and poorer response to treatment than did the dietary subtype. DISCUSSION: Results suggest that moderate dieting is a central feature of BED and that affective disturbances occur in only a subset of cases. However, the confluence of dieting and negative affect signals a more severe variant of the disorder marked by elevated psychopathology, impaired social functioning, and a poorer treatment response. PMID- 11439406 TI - Body image and personality predictors of eating disorder symptoms during the college years. AB - OBJECTIVE: Women entering college (N = 118) were longitudinally followed for 3 years with assessments of eating pathology (Restraint and Bulimia). METHOD: Measures taken at Time 1 included timing of onset of puberty, Figure Dissatisfaction, Ineffectiveness, Public Self-Consciousness, and mood states (Profile of Mood States). Measures were evaluated as predictors of changes in Restraint and Bulimia scores across the three-year period. RESULTS: Both Restraint (r = .69) and Bulimia (r = .50) were quite stable across the 3 years. Mean weight gain of 5.4 pounds (p < .001) was paralleled by an increase in "ideal weight" of 4.7 pounds. Hierarchical regressions controlling for Time 1 levels of eating disorder symptoms revealed that changes in Bulimia (R(2) = .38, p < .0001) were related to the psychological measures and to Figure Dissatisfaction. Changes in Restraint (R(2) = .61, p < .0001) were significantly related to Figure Dissatisfaction. DISCUSSION: Findings are discussed in the context of the value of longitudinal designs in identifying risk factors. Dissatisfaction with one's figure seems to be consistently related to worsening eating pathology. PMID- 11439407 TI - Co-occurring eating and depressive problems: an 8-year study of adolescent girls. AB - OBJECTIVE: This paper examines prospectively the co-occurrence of eating and depressive problems in 105 White girls who were seen at three times from young adolescence to young adulthood. METHOD: Girls were from middle to upper-middle class families. Co-occurrence of eating and depressive problems was determined cross-sectionally from questionnaire data using established criteria for identifying subclinical problems. RESULTS: The rate of depressive problems declines across middle to late adolescence while the rate of eating problems is fairly constant across all three times of assessment. Analyses suggest that girls with depressive problems (with and without co-occurring eating problems) experience impairments in peer and family relationships; girls with high scores on both problems have poor adjustment across several domains. DISCUSSION: The additional psychological strains seen with co-occurrence of eating and depressive problems heighten physical and mental health concomitants of both problems across adolescence. PMID- 11439408 TI - Relationship between depression and body dissatisfaction in women diagnosed with bulimia nervosa. AB - OBJECTIVE: Body dissatisfaction and depression have consistently demonstrated a positive association in women. This study sought to determine the independence of this association from bulimic symptomatology among women diagnosed with bulimia nervosa. METHOD: Participants were 101 women who completed a controlled treatment study of bulimia nervosa and participated in follow-up assessments 10 years later. RESULTS: Findings indicated that baseline levels of depression were independent of and superior to bulimic symptoms in prospectively predicting body dissatisfaction at follow-up assessment. DISCUSSION: Findings suggest that depression may be a better prognostic indicator of body dissatisfaction than bulimic symptoms in women diagnosed with bulimia nervosa. A model in which depression represents a contributing factor for the maintenance of body dissatisfaction is discussed. PMID- 11439409 TI - Altered response to meta-chlorophenylpiperazine in anorexia nervosa: support for a persistent alteration of serotonin activity after short-term weight restoration. AB - OBJECTIVE: Patients with anorexia nervosa (AN) have disturbances of appetite and behaviors, such as dysphoria, inhibition, and obsessions, that could be related to altered serotonin activity. To investigate such relationships, we administered meta-chlorophenylpiperazine (m-CPP), a relatively serotonin-specific drug. METHODS: To avoid the confounding effects of malnutrition or weight loss, we studied 12 patients with restricting-type AN between 5 and 17 days after a return to a normal weight and while on a stable dietary intake. We compared them to 12 healthy control women (CW). m-CPP was administered double blind and placebo controlled. RESULTS: Although weight restored, AN women had lower body weight and increased ratings for depression and obsessionality compared with CW. After m CPP, AN women had an elevation in mood and a reduction in body image distortion when compared with placebo. After m-CPP, groups had similar cortisol, adrenocorticotropin (ACTH), and growth hormone responses whereas AN women had an uncertain reduction in prolactin response. DISCUSSION: These data support other studies that suggest that altered serotonin activity persists after weight restoration in AN patients. The finding that m-CPP temporarily improved mood and reduced body image distortions supports the hypothesis that altered serotonin activity may contribute to the pathophysiology of AN. PMID- 11439410 TI - Changing patterns of hospitalization in eating disorder patients. AB - OBJECTIVE: This study investigated the changing patterns of hospitalization of eating disorder patients over the past 15 years. METHOD: The records of 1,185 eating disorder patients between 1984 and 1998 were examined on several variables. RESULTS: Over the 15 years, the number of first admissions increased from 20 to 182. There was a concomitant decrease in length of stay from 149.5 days in 1984 to 23.7 days in 1998. Readmissions increased markedly from 0% during the first year to 27% of total admissions in 1998. The discharge weight of anorectic patients significantly decreased from a body mass index (BMI) of 19.3 in 1984 to 17.7 in 1998. These changes were particularly salient in the past 3 years, concurrent with a dramatic rise in managed care cases. CONCLUSIONS: Over the past 15 years, eating disorder hospital treatment has metamorphozed from long term treatment of a disorder to stabilization of acute episodes. For some patients, this change has been deleterious and not cost effective. PMID- 11439411 TI - Effects of a circuit weight training program on the body images of college students. AB - OBJECTIVE: The present investigation examined the extent to which participation in a 6-week circuit-weight training program produced changes in participants' body images relative to a matched control group. METHOD: The weight trainers consisted of 39 college students (27 women and 12 men). The control group of 39 individuals did not weight-train currently or within the past year. All participants were pre- and posttested on the Multidimensional Body Self-Relations Questionnaire, the Social Physique Anxiety Scale, and the Physical Self-Efficacy Scale. Weight trainers were also pre- and posttested on muscular strength and assessed on their motives for exercise. RESULTS: The program successfully increased upper- and lower-body strength. In contrast to the comparison group, weight trainers had a significantly improved evaluation of their appearance, greater body satisfaction, reduced social physique anxiety, and enhanced physical self-efficacy. Outcomes were unrelated to the extent of concurrent aerobic exercise and largely unrelated to exercise motives. DISCUSSION: Even a relatively brief weight training program can produce improvements in multiple aspects of body image. Further research should investigate weight training as an adjunct to psychosocial treatments of body dissatisfaction. PMID- 11439412 TI - The use of ecological momentary assessment approaches in eating disorder research. AB - OBJECTIVE: Eating disorder (ED) research is increasingly focusing on the proximal antecedents to disordered eating behavior. Such antecedents may include cognitions, environmental stimuli, social interactions, and affective states. Current ED theories suggest that the relationships between antecedents and eating behavior may be complex, including interaction associations, time-lagged effects, and associations that persist only for brief periods of time. Similarly, these theories often include the consequences of behavior-influencing variables of interest (e.g., short-term reductions in negative affect). Careful examination of such theories, however, has been limited by a reliance on data collection methods not appropriate for testing these effects. METHOD: This study examines alternative methods for data collection and analysis that overcome previously noted limitations, using data collected in several studies with eating-disordered participants. RESULTS: The development of a technique called ecological momentary assessment (EMA) allows the ongoing study of behavior in its natural context and reduces biases associated with retrospective recall. The development of technology that allows the sophisticated collection and storage of such data (e.g., palm-top computers), along with statistical procedures for analyzing hierarchically nested, repeated measures data, allow precise testing of complex theoretical models. DISCUSSION: We demonstrate several important features of this research: (1) patients are willing and able to engage in EMA studies, (2) data not possible to collect using other designs are obtainable, (3) complex theoretical models can be evaluated using these data and appropriate statistical methods, and (4) the collection and analysis of EMA data present unique difficulties to ED researchers. Finally, we endorse and provide recommendations for the use of EMA in future ED research and practice. PMID- 11439413 TI - Stage of change as a predictor of response to psychotherapy for bulimia nervosa. AB - OBJECTIVE: This study was designed to test the hypotheses that stage of change is a useful predictor of dropout and that it is related to treatment outcome in patients receiving brief psychotherapy for bulimia nervosa. METHOD: One hundred ten patients with bulimia nervosa were randomly assigned treatment with cognitive behavioral therapy (CBT) or interpersonal therapy (IPT). On initial screening, patients were classified as being in the stage of precontemplation, contemplation, or preparation. RESULTS: Initial stage of change was not related to dropout either before or following randomization. Among all treatment completers, stage of change was related to outcome. Of the patients assigned to CBT, stage of change was not related to outcome. However, stage of change was related to outcome in patients randomized to IPT. DISCUSSION: This study suggests that initial stage of change may, under certain circumstances, be a useful predictor of outcome and that its utility as an outcome predictor may vary according to treatment. PMID- 11439414 TI - Dialectical behavior therapy adapted for bulimia: a case report. AB - OBJECTIVE: This case report describes the application of dialectical behavior therapy (DBT) to the treatment of bulimia nervosa in a 20-session manualized therapy. METHOD: The treatment, based on an affect regulation model of eating disorders, was developed to teach emotion regulation skills to replace eating disordered behaviors. The patient, a 36-year-old woman, had a long history of binge eating and purging that had not responded to 2 years of counseling. In the 4 weeks before treatment began, she reported 13 objective binges and 21 purging episodes. RESULTS AND DISCUSSION: Upon initiating DBT, her binge eating and purging rapidly declined. She achieved abstinence by the fifth week of treatment and maintained it through treatment. In the 6 months following treatment, she reported a total of two objective binge episodes and two purge episodes. PMID- 11439415 TI - Binge eating disorder onset by unusual parasitic intestinal disease: a case report. AB - We report a case of binge eating disorder (BED) in a 19-year-old Spanish woman, whose onset followed a nowadays uncommon parasitic intestinal disease (Taenia solium by cysticercosis). The patient exhibited bizarre and frequent hyperphagia episodes and extreme loss of weight, common symptoms of this condition. The patient continued to have frequent binge episodes and inappropriate eating patterns, gaining weight consequently over a normal range, despite successful treatment of the condition. No purging behavior was detected. A classical conditioning behavioral model, was useful for the understanding and formulation of this case. To the authors' knowledge, this is the first reported case where a parasitic infestation triggered the onset of BED. PMID- 11439416 TI - Simultaneous bilateral spontaneous pneumothoraces in a young woman with anorexia nervosa. AB - An eighteen year-old woman with anorexia nervosa was admitted to hospital with acute dyspnea. There was no history of trauma to the chest and the patient denied any vomiting. METHOD: A chest radiograph was taken which showed bilateral pneumothoraces with complete collapse of both lungs. RESULTS: Following insertion of bilateral chest drains the patient made a complete recovery, later having bilateral pleurectomies performed to prevent recurrence. DISCUSSION: We discuss the evidence suggesting that the state of malnutrition that results from anorexia nervosa may in fact predispose patients to the development of pneumothoraces. PMID- 11439417 TI - Aggravation of food-related behavior in an adolescent with Prader-Willi syndrome treated with fluvoxamine and fluoxetine. AB - Prader-Willi Syndrome is a congenital multisystem disorder, characterized by a typical dysmorphism, mental retardation, hyperphagia due to insatiable appetite, and resultant morbid obesity. Psychiatric symptoms include obsessions and temper tantrums. METHOD: Pharmacotherapy is experimental with a few reports of successful fluoxetine treatment. RESULTS: We report an aggravation in the food related symptoms and a consequent weight gain in an adolescent with Prader-Willi syndrome, who was treated with fluvoxamine and fluoxetine. PMID- 11439418 TI - Can steroid use be a precipitant in the development of an eating disorder? AB - OBJECTIVE: Corticosteroids have a wide range of clinical indications in the treatment of both acute and chronic medical illnesses, and weight gain is a well documented side effect of their use. In this paper, we describe eight individuals with an eating disorder, which appeared following steroid administration for a medical condition. METHOD: These findings, support the possibility that the excessive weight gain resulting from steroid use can be a precipitating factor in the development of an eating disorder, particularly among female adolescents and young women who are preoccupied with their appearance and weight. RESULTS: Physicians caring for women receiving corticosteroids should therefore familiarize themselves with these patients' pre-morbid and current eating habits, as well as their weight, dieting and body image histories. DISCUSSION: The relevance of these cases for furthering our understanding of the development and treatment of eating disorder is also discussed. PMID- 11439419 TI - Can you ever be too old to be too thin? Anorexia nervosa in a 92-year-old woman. AB - Anorexia nervosa is a common psychiatric condition that most often affects young adults. If left untreated, it can have significant morbidity and mortality. METHODS AND DISCUSSION: In this report, we describe a woman first diagnosed with anorexia nervosa at age 92 and we discuss the relevant diagnostic and treatment related issues. CONCLUSION: This case underscores the need to heighten diagnostic sensitivity for this and other eating disorders at any age. PMID- 11439420 TI - Predicting analysis times in randomized clinical trials. AB - Randomized clinical trial designs commonly include one or more planned interim analyses. At these times an external monitoring committee reviews the accumulated data and determines whether it is scientifically and ethically appropriate for the study to continue. With failure-time endpoints, it is common to schedule analyses at the times of occurrence of specified landmark events, such as the 50th event, the 100th event, and so on. Because interim analyses can impose considerable logistical burdens, it is worthwhile predicting their timing as accurately as possible. We describe two model-based methods for making such predictions during the course of a trial. First, we obtain a point prediction by extrapolating the cumulative mortality into the future and selecting the date when the expected number of deaths is equal to the landmark number. Second, we use a Bayesian simulation scheme to generate a predictive distribution of milestone times; prediction intervals are quantiles of this distribution. We illustrate our method with an analysis of data from a trial of immunotherapy in the treatment of chronic granulomatous disease. PMID- 11439421 TI - The effects of outcome misclassification and measurement error on the design and analysis of therapeutic equivalence trials. AB - In any clinical trial, the use of imperfect diagnostic procedures or laboratory techniques may lead to misclassification and measurement error in the primary outcome. Although the effects of non-differential outcome misclassification and measurement error on conventional superiority trials have been extensively investigated, less is known about the impact of these errors on the results and interpretation of therapeutic equivalence trials. In this paper we formally investigate the effects of outcome misclassification and measurement error on the estimates of treatment effects, type I error rate, and power of equivalence trials. Our results indicate that, contrary to what one may expect based on the well known attenuating effects of non-differential error in conventional studies, these errors do not always favour the goal of demonstrating equivalence. The magnitude and direction of the influence depend on a number of factors including the nature of the outcome variable, specific formulation of equivalence, size of the error rates, and assumptions regarding the true treatment effect. PMID- 11439422 TI - Patient specific dosing in a cancer phase I clinical trial. AB - Recent improvements in our understanding of drug metabolism have led to the development of anticancer therapies that accommodate patient differences in drug tolerance. Such methods adjust the dose level according to measurable patient characteristics in order to obtain a target drug exposure. This paper describes the utilization of a patient specific dosing scheme in the statistical design of a phase I clinical trial involving patients with advanced adenocarcinomas of gastrointestinal origin. During the trial, dose levels were adjusted according to each patient's pretreatment concentration of an antibody that was shown in preclinical testing to moderate the effect of the agent under investigation. The design of the trial permitted a continual adjustment of the model used to tailor the dose to each patient's individual needs. PMID- 11439423 TI - Projecting the standard error of the Kaplan-Meier estimator. AB - Clinical studies in which a major objective is to produce Kaplan-Meier estimates of survival probabilities should be designed to produce those estimates with a desired prespecified precision as measured by their standard errors. By considering the Peto and Greenwood formulae for the estimated standard error of the Kaplan-Meier estimate and replacing their constituents with expected values based on the study's design parameters, formulae for projected standard errors can be produced. These formulae are shown, through simulations, to be quite accurate. PMID- 11439424 TI - Relative risk estimation and inference using a generalized logrank statistic. AB - When comparing two survival distributions with proportional hazard functions, the logrank test is optimal for testing the null hypothesis that the constant hazard ratio (relative risk) is one. In this paper, we focus on (i) testing for departures from a relative risk other than one, and (ii) estimation of the relative risk. The standard tool to address both (i) and (ii) is the Cox proportional hazards model. However, the performance of the Cox model can be less than optimal with small samples. We show why this is the case, and propose a simple alternative method of estimation and inference based on a generalized logrank (GLR) statistic. While the GLR and Cox model approaches are asymptotically similar, empirical results reveal that the GLR approach is notably more efficient than the Cox model when the number of subjects is small (< 100 subjects per treatment group). An example based on survival times of cervical cancer patients is used to illustrate the proposed methodology. PMID- 11439425 TI - Two-stage methods for the analysis of pooled data. AB - Epidemiologic studies of disease often produce inconclusive or contradictory results due to small sample sizes or regional variations in the disease incidence or the exposures. To clarify these issues, researchers occasionally pool and reanalyse original data from several large studies. In this paper we explore the use of a two-stage random-effects model for analysing pooled case-control studies and undertake a thorough examination of bias in the pooled estimator under various conditions. The two-stage model analyses each study using the model appropriate to the design with study-specific confounders, and combines the individual study-specific adjusted log-odds ratios using a linear mixed-effects model; it is computationally simple and can incorporate study-level covariates and random effects. Simulations indicate that when the individual studies are large, two-stage methods produce nearly unbiased exposure estimates and standard errors of the exposure estimates from a generalized linear mixed model. By contrast, joint fixed-effects logistic regression produces attenuated exposure estimates and underestimates the standard error when heterogeneity is present. While bias in the pooled regression coefficient increases with interstudy heterogeneity for both models, it is much smaller using the two-stage model. In pooled analyses, where covariates may not be uniformly defined and coded across studies, and occasionally not measured in all studies, a joint model is often not feasible. The two-stage method is shown to be a simple, valid and practical method for the analysis of pooled binary data. The results are applied to a study of reproductive history and cutaneous melanoma risk in women using data from ten large case-control studies. PMID- 11439426 TI - A generalized concordance correlation coefficient for continuous and categorical data. AB - This paper discusses a generalized version of the concordance correlation coefficient for agreement data. The concordance correlation coefficient evaluates the accuracy and precision between two measures, and is based on the expected value of the squared function of distance. We have generalized this coefficient by applying alternative functions of distance to produce more robust versions of the concordance correlation coefficient. In this paper we extend the application of this class of estimators to categorical data as well, and demonstrate similarities to the kappa and weighted kappa statistics. We also introduce a stratified concordance correlation coefficient which adjusts for explanatory factors, and an extended concordance correlation coefficient which measures agreement among more than two responses. With these extensions, the generalized concordance correlation coefficient provides a unifying approach to assessing agreement among two or more measures that are either continuous or categorical in scale. PMID- 11439427 TI - Evaluation of an adjusted chi-square statistic as applied to observational studies involving clustered binary data. AB - A simple adjustment to the Pearson chi-square test has been proposed for comparing proportions estimated from clustered binary observations. However, the assumptions needed to assure the validity of this test have not yet been thoroughly addressed. These assumptions will hold for experimental comparisons, but could be violated for some observational comparisons. In this paper we investigate the conditions under which the adjusted chi-square statistic is valid and examine its performance when these assumptions are violated. We also introduce some alternative test statistics that do not require these assumptions. The test statistics considered are then compared through simulation and an example presented based on real data. The simulation study shows that the adjusted chi-square statistic generally produces empirical type I errors close to nominal under the assumption of a common intracluster correlation coefficient. Even if the intracluster correlations are different, the adjusted chi-square statistic performs well when the groups have equal numbers of clusters. PMID- 11439428 TI - On determination of sample size in hierarchical binomial models. AB - We consider a two- and a three-stage hierarchical design containing the effects of k clusters with n units per cluster. In the two-stage model, the conditional distribution of the discrete response Y(i) is assumed to be independent binomial with mean n(straight theta)i (I=1,....k). The success probabilities, straight theta(i)'s, are assumed exchangeable across the k clusters, each arising from a beta distribution. In the three-stage model, the parameters in the beta distribution are assumed to have independent gamma distributions. The size of each cluster, n, is determined for functions of straight theta(i). Lengths of central posterior intervals are computed for various functions of the straight theta(i)'s using Markov chain Monte Carlo and Monte Carlo simulations. Several prior distributions are characterized and tables are provided for n with given k. Methods for sample size calculations under the two- and three-stage models are illustrated and compared for the design of a multi-institutional study to evaluate the appropriateness of discharge planning rates for a cohort of patients with congestive heart failure. PMID- 11439429 TI - Disease map reconstruction. AB - The analysis of the geographical distribution of disease incidence or prevalence is now of considerable importance for public health workers and epidemiologists alike. Important disease variations often have a spatial expression and so spatial analysis methods are an important additional tool in this connection. In this tutorial I have aimed to highlight the main issues relating to the analysis of disease where the goal is the reduction in noise in a disease map. This area is sometimes simply called disease mapping. A number of modelling approaches to disease mapping are considered and a case study highlighting the methods advocated is also included. PMID- 11439430 TI - Statistics in medical journals: some recent trends. PMID- 11439431 TI - An approximate unconditional test of non-inferiority between two proportions. PMID- 11439433 TI - How risky is heart-valve replacement? PMID- 11439434 TI - Ask the doctor. I recently found out I have atrial fibrillation. My doctor wants me to take a blood thinner for a while and then have cardioversion. But I'll be away from home for a few months, so I'd like to hold off on the cardioversion until I get back. My doctor doesn't want me to delay. Why can't it wait? PMID- 11439435 TI - By the way, doctor... I don't know whether to start hormone-replacement therapy. I'm having a terrible time with hot flashes, but I'm worried about the risk for ovarian and breast cancer. What should I do? PMID- 11439436 TI - Depression revisionism. PMID- 11439437 TI - Sunscreens, sunburn, and skin cancer. PMID- 11439438 TI - [Amyloidosis: when to consider it? How to confirm it?]. PMID- 11439439 TI - [Hereditary amyloidosis]. PMID- 11439440 TI - [AL amyloidosis and monoclonal immunoglobulin deposit diseases]. PMID- 11439441 TI - [Treatment of AL amyloidosis]. PMID- 11439443 TI - [From the protein to amyloid deposit]. PMID- 11439444 TI - The effects of bilateral lesions of the amygdala on dyadic social interactions in rhesus monkeys (Macaca mulatta). AB - The role of the amygdala in dyadic social interactions of adult rhesus monkeys (Macaca mulatta) was assessed after bilateral ibotenic acid lesions. Social, nonsocial, and spatial behaviors of amygdalectomized and control monkeys were assessed in 3 dyadic experiments: constrained, unconstrained, and round robin. Lesions produced extensive bilateral damage to the amygdala. Across all experiments, the amygdalectomized monkeys demonstrated increased social affiliation, decreased anxiety, and increased confidence compared with control monkeys, particularly during early encounters. Normal subjects also demonstrated increased social affiliation toward the amygdalectomized subjects. These results indicate that amygdala lesions in adult monkeys lead to a decrease in the species normal reluctance to immediately engage a novel conspecific in social behavior. The altered behavior of the amygdalectomized monkeys may have induced the increased social interactions from their normal companions. This is contrary to the idea that amygdalectomy produces a decrease in social interaction and increased aggression from conspecifics. PMID- 11439445 TI - Effects of selective neonatal temporal lobe lesions on socioemotional behavior in infant rhesus monkeys (Macaca mulatta). AB - Normal infant monkeys and infant monkeys with neonatal damage to either the medial temporal lobe or the inferior temporal visual area were assessed in dyadic social interactions at 2 and 6 months of age. Unlike the normal infant monkeys, which developed strong affiliative bonds and little or no behavioral disturbances, the lesioned monkeys (each of which was observed with an unoperated control) exhibited socioemotional abnormalities and aberrant behaviors. The socioemotional changes predominated at 6 months of age and were particularly severe in monkeys with medial temporal lesions. In both the pattern and time course, the socioemotional deficits produced by the neonatal medial temporal lesions bear a striking resemblance to the behavioral syndrome in children with autism. Further analysis of these lesion-induced abnormalities in nonhuman primates may therefore provide insight into this debilitating human developmental disorder. PMID- 11439446 TI - Impaired delay eyeblink classical conditioning in individuals with anterograde amnesia resulting from anterior communicating artery aneurysm rupture. AB - Anterior communicating artery (ACoA) aneurysm rupture can lead to an anterograde amnesia syndrome similar to that observed after damage to the hippocampus and medial temporal lobes (MT). It is currently believed that ACoA amnesia results from basal forebrain damage that disrupts hippocampal processing without direct hippocampal damage. Converging evidence from animal studies and computational modeling suggests that qualitative differences may exist in the pattern of memory impairment after basal forebrain or MT damage. For example, animals with basal forebrain but not hippocampal damage are impaired at delay eyeblink classical conditioning (EBCC). In this study, individuals with ACoA amnesia were shown to be impaired at delay EBCC compared with matched controls; this contrasts with the spared delay EBCC previously observed in MT amnesia. This finding suggests the beginning of a possible dissociation between the memory impairments in MT versus ACoA amnesia. PMID- 11439447 TI - Angular velocity and head direction signals recorded from the dorsal tegmental nucleus of gudden in the rat: implications for path integration in the head direction cell circuit. AB - When a rat navigates through space, head direction (HD) cells provide an ongoing signal of the rat's directional heading. It is thought that these cells rely, in part, on angular path integration of the rat's head movements. This integration requires that the HD cell system receive information about angular head movements and that this information be combined with the current directional signal, to generate the next "predicted" direction. Recent data suggest that the dorsal tegmental nucleus (DTN) may play a critical role in helping to generate the HD cell signal. To test this, recordings were made from cells in the DTN in freely moving rats. The following cell types were found: (a) "classic" HD cells, (b) angular velocity cells, and (c) cells that fired as a function of both head direction and angular velocity. Thus, DTN cells exhibit firing characteristics that are critical to the neural circuit hypothesized for generation of the HD cell signal. PMID- 11439448 TI - Cognitive task performance after lidocaine-induced inactivation of different sites within the basolateral amygdala and dorsal striatum. AB - To determine whether discrete components of amygdaloid and striatal memory systems could interact to guide behavior in a radial arm maze, conditioned cue preference (CCP) and win-stay accuracy were examined after lidocaine inactivation of either the rostral (rBLA) or caudal (cBLA) basolateral amygdala, the lateral (lDST) or medial (mDST) dorsal striatum, or a control site in rats. CCP expression was blocked only after rBLA or cBLA inactivation. lDST inactivation prevented attainment of criteria win-stay performance, whereas rBLA and mDST inactivation delayed it. Control site inactivation did not influence performance in either task. These findings suggest that the amygdala works independently of other memory systems to regulate learned responses in the CCP task, the rBLA may work cooperatively with the lDST to guide behavior in the win-stay task, and the mDST is less critical than the lDST for attaining criteria performance in the win stay task. PMID- 11439449 TI - Encoding of geometric and landmark information in the left and right hemispheres of the Avian Brain. AB - Chicks were trained binocularly to find food buried under sawdust in the center of a square enclosure. When tested in an enclosure made larger or smaller in size, binocular and left-eyed chicks searched mainly on the basis of relative distance of the food from the enclosure walls, whereas right-eyed chicks searched on the basis of absolute distance. Moreover, binocular and left-eyed chicks relied mainly on global spatial information (i.e., distances from the walls), whereas right-eyed chicks also used information provided by visual landmarks. These results suggest that the right hemisphere of the avian brain (fed mostly by the left eye) is primarily concerned with encoding of relational spatial information, whereas the left hemisphere (fed mainly by the right eye) is concerned with absolute metric information, possibly as part of an encoding strategy based primarily on local (both spatial and nonspatial) cues. PMID- 11439450 TI - Hippocampal growth and maintenance depend on food-caching experience in juvenile mountain chickadees (Poecile gambeli). AB - This experiment investigated the development of caching behavior and the hippocampus (HF) in postfledging mountain chickadees (Poecile gambeli). From Days 35 to 53, the number of seeds stored increased but the proportion recovered did not. Birds that stored and recovered during 3 or more trials had significantly enlarged HF but not telencephalon volumes (experienced) compared with those that stored but did not recover (store only) and those deprived of caching experience altogether (deprived). HF size did not increase linearly with the number of experience trials. Birds that received less than 3 experience trials did not differ from deprived birds in HF size, suggesting a threshold effect. Experienced birds prevented from caching for 1 month had significantly smaller HF volumes than those examined immediately after caching experience and did not differ from deprived birds. Experience of both storing and recovery is required to initiate growth and maintain HF size. PMID- 11439451 TI - OMP gene deletion results in an alteration in odorant quality perception. AB - To test the hypothesis that odorant quality perception is altered in olfactory marker protein (OMP)-null mice, we trained and tested adult OMP-null and control mice, using a 5-odorant identification confusion matrix task (animal odorant confusion matrix [AOCM]). On average, control and null mice performed the task at equivalent levels. The composite 5 x 5 response matrix from 40 testing sessions for each subject (both OMP-null and control) was compared with that of every other subject, yielding a dissimilarity matrix of AOCM responses. A multidimensional scaling (MDS) analysis of the dissimilarity data yielded a 4 dimensional solution, with each mouse occupying a point in MDS animal space. Statistical analysis demonstrated significant effects of genotype in determining the location of a mouse in the MDS space. These data suggest, therefore, that compared with that of controls, odorant quality perception is altered in the OMP null mouse. PMID- 11439452 TI - Enantioselectivity of odor perception in honeybees (Apis mellifera carnica). AB - The authors tested the ability of 60 free-flying honeybees (Apis mellifera carnica) to discriminate a conditioning odor from an array of 26 simultaneously presented substances. The stimuli included 10 pairs of enantiomers and 6 essential oils. The bees (a) significantly distinguished between 98% of the 540 odor pairs tested, thus showing an excellent overall discrimination performance, and (b) were able to discriminate between the optical isomers of limonene, alpha pinene, beta-citronellol, menthol, and carvone but failed to distinguish between the (+)- and (-)-forms of alpha-terpineol, camphor, rose oxide, fenchone, and 2 butanol. The findings support the assumptions that enantioselective molecular odor receptors may exist only for some volatile enantiomers and that insects and mammals may share common principles of odor quality perception, irrespective of their completely differing repertoires of olfactory receptors. PMID- 11439453 TI - A mutation in the AMPA-type glutamate receptor, glr-1, blocks olfactory associative and nonassociative learning in Caenorhabditis elegans. AB - The alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA)-type ionotropic glutamate receptor mediates fast excitatory neurotransmission in the vertebrate brain and is important for synaptic plasticity and the initial induction of long term potentiation (LTP). This study found that the putative Caenorhabditis elegans AMPA receptor gene, glr-1, plays a significant role in experience dependent behavior in C. elegans. glr-1 mutants are deficient in an olfactory associative learning task, in which diacetyl (DA) is paired with acetic acid solution. glr-1 mutant nematodes are also impaired in nonassociative learning (habituation) with the same DA stimulus. The C. elegans learning mutants, lrn-1 and lrn-2, are impaired in chemosensory associative learning yet have no deficits in habituation. The results suggest that although associative and nonassociative learning can be genetically dissociated (lrn-1 and lrn-2), they also share some common molecular processes, including glr-1-mediated neurotransmission. PMID- 11439454 TI - Studies of binaural detection in the rabbit (Oryctolagus cuniculus) with Pavlovian conditioning. AB - A Pavlovian conditioned eyeblink response in rabbits (Oryctolagus cuniculus) was used to study psychoacoustical phenomena previously demonstrated in human listeners and other animals. This article contains the results of a tone-in-noise detection study to examine 2 psychoacoustical phenomena in rabbit and in human listeners: (a) the binaural masking level difference (BMLD) and (b) differential performance across reproducible noise masker waveforms. The rabbits demonstrated a BMLD comparable in size to other species. Significant differences in performance across reproducible noise masker waveforms were seen in the rabbits. This performance was compared with the performance of human listeners using the same set of waveforms. PMID- 11439455 TI - Learning from others to cope with biting flies: social learning of fear-induced conditioned analgesia and active avoidance. AB - Although fear conditioning has received extensive attention, little is known about the roles of social learning whereby an individual may learn and acquire the fear responses of another. The authors examined individually and socially mediated acquisition of fear and analgesia to the natural aversive stimulus of biting flies. Exposure to biting flies elicited in individual naive mice analgesia and active self-burying to avoid the flies. When exposed 24 hr later to flies whose biting parts were removed, but not to nonbiting house flies, these mice displayed conditioned analgesia and self-burying. This "one-trial" conditioned analgesia and avoidance was also acquired through social learning without direct individual experience with biting flies. Naive "observer" mice that witnessed other "demonstrator" mice being attacked by biting flies exhibited analgesia and self-burying 24 hr later to altered flies. PMID- 11439456 TI - Effects of warmth on newborn rats' motor activity and oral responsiveness to an artificial nipple. AB - Temperature is a powerful regulator of the behavior and physiology of newborn altricial animals. The effects of warmth on newborn rats' oral responsiveness to suckling stimuli and spontaneous motor activity in a thermoneutral environment were investigated. Newborn rat pups' oral grasp responses to an artificial nipple and overall motor activity were recorded for 18 min. Near-term pups were delivered by cesarean section so that their 1st experiences with suckling stimuli could be observed. Experimental pups were warmed for 15 s every 2 min; control pups were not warmed. Warmed pups grasped the nipple fewer times than the not warmed pups. However, oral grasp durations became longer for the warmed pups but not for the not-warmed pups. Warmth increased pups' motor activity but only while the heat was applied. Warmth in a thermoneutral environment may promote longer nipple attachment during newborns' early feeding experiences. PMID- 11439457 TI - Comparison of two positive reinforcing stimuli: pups and cocaine throughout the postpartum period. AB - This set of experiments investigated the appetitive or motivational processes underlying the performance of maternal behavior. The place preference paradigm was adapted to simultaneously investigate the reinforcing properties of cocaine and pups for maternal, lactating dams. These modifications allowed the authors to assess which stimulus, either a 10 mg/kg s.c. injection of cocaine or 3 pups, had the strongest reinforcing value. At Postpartum Days 10 and 16, the dams preferred the cocaine cue-associated chamber, whereas the dams tested at Postpartum Day 8 preferred the pup cue-associated chamber. Overall, the data revealed an interaction between the postpartum period at testing and the exhibited preference for cocaine or pups. Further testing will investigate the neural circuitry underlying the appetitive processes of each stimulus. PMID- 11439458 TI - Transient sex differences in the between-sessions but not in the within-session memory underlying an active place avoidance task in weanling rats. AB - Spatial abilities were tested in male and female rats by training them to avoid an area in which there was a mild footshock while the arena rotated at 1 revolution/minute. The to-be-avoided area was stable in the coordinates of the room, so extramaze landmarks had to be used for accurate navigation, as the rotation made intramaze cues and substrate-based path integration useless for the avoidance. From Postnatal Day (PD) 19, rats were trained for 22 consecutive days. When the shock area was the same across sessions male rats reached optimal performance on PDs 23-24, 10 days before female rats, but when the location of the shock changed daily there were no sex differences. The results indicate that there are separate memory components underlying spatial competence: a within session component that develops similarly in male and female rats and a between sessions component that lasts at least 24 hr and appears earlier in male than in female rats. PMID- 11439459 TI - The role of cholecystokinin in conditional compensatory responding and morphine tolerance in rats. AB - As elaborated in the conditioning analysis of tolerance, cues present at the time of drug administration become associated with the drug effect. A particularly salient cue that may become associated with the drug effect is the pharmacological drug-onset cue inherent to drug administration. Drug-associated cues contribute to tolerance by eliciting a conditional compensatory response that attenuates the drug effect. For example, the early drug effect, having been paired with the subsequent larger drug effect, may elicit the release of antiopioid peptides that counter opioid effects. The role of a putative antiopioid peptide, cholecystokinin-8 (CCK), in the associative mechanisms of opiate tolerance was evaluated. The results of these experiments suggest that a CCK2 receptor antagonist attenuates both the expression of opiate tolerance and the conditional compensatory response hypothesized to mediate such tolerance. PMID- 11439460 TI - Conditioned brain-stimulation reward attenuates the acoustic startle reflex in rats. AB - The acoustic startle reflex (ASR) in rats is attenuated by a light paired with food or, in humans, by "pleasant" pictures. Rats were trained to barpress for lateral hypothalamus (LH) stimulation. ASR amplitudes were then measured at 4 intensities, with or without a light. Control rats that did not receive brain stimulation reward (BSR) showed initially lower ASR amplitudes than did rats exposed to BSR, but both groups responded similarly with or without light. Next, experimental rats were given BSR in the presence of light but not in its absence. After conditioning, ASR amplitudes were reduced, and ASR thresholds were raised by a mean of 2.6 dB in the light but remained at preconditioning levels without light. No such change was found for control rats or rats with placements outside the LH. PMID- 11439461 TI - The role of drinking in the suppression of food intake by recent activity. AB - The standard activity-based anorexia procedure provides rats with access to a running wheel while restricting their access to dry food. This can produce reduced food intake and progressive weight loss. Using this procedure, in the present study (Experiment 1) the authors found changes in drinking patterns both in the period of high activity preceding food access and during the feeding period. Varying the procedure by providing wet mash (Experiment 2) or by prior adaptation to a drinking schedule (Experiment 3) prevented the self-starvation effect. These results indicate the importance of drinking when analyzing the effect of recent activity on food intake. PMID- 11439462 TI - Eyeblink conditioning in the rabbit (Oryctolagus cuniculus) with stimulation of the mystacial vibrissae as a conditioned stimulus. AB - Eyeblink conditioning is a well-understood paradigm for the study of learning and memory and has been successfully employed with the use of auditory and visual conditioned stimuli (CSs). In this study, vibrotactile stimulation of the mystacial vibrissae was examined as an alternative CS in the rabbit (Oryctolagus cuniculus). The technique is described and acquisition of eyeblink conditioning (EBC) with stimulation of a single row of vibrissae in a delay paradigm is reported. Extinction of EBC with presentation of the CS alone is demonstrated, as well as reacquisition with stimulation of a single whisker. Finally, control experiments ensure that the CS has no auditory components. Ipsilateral presentation of the CS and airpuff is a more effective combination for training than contralateral presentations. Vibrotactile stimulation of the vibrissae as a CS will enable further examination of the neural correlates of learning in a well characterized sensory system. PMID- 11439463 TI - Theory and practice in the design of physician payment incentives. AB - Combining the economic literature on principal-agent relationships with examples of marketplace innovations allows analysis of the evolution of methods for paying physicians. Agency theory and the economic principles of performance-based compensation are applied in the context of imperfect information, risk aversion, multiple interrelated tasks, and team production efficiencies. Fee-for-service and capitation are flawed methods of motivating physicians to achieve specific goals. Payment innovations that blend elements of fee-for-service, capitation, and case rates can preserve the advantages and attenuate the disadvantages of each. These innovations include capitation with fee-for-service carve-outs, department budgets with individual fee-for-service or "contact" capitation, and case rates for defined episodes of illness. The context within which payment incentives are embedded, includes such non-price mechanisms as screening and monitoring and such organizational relationships as employment and ownership. The analysis has implications for health services research and public policy with respect to physician payment incentives. PMID- 11439464 TI - Partnership synergy: a practical framework for studying and strengthening the collaborative advantage. AB - The substantial interest and investment in health partnerships in the United States is based on the assumption that collaboration is more effective in achieving health and health system goals than efforts carried out by single agents. A clear conceptualization of the mechanism that accounts for the collaborative advantage, and a way to measure it are needed to test this assumption and to strengthen the capacity of partnerships to realize the full potential of collaboration. The mechanism that gives collaboration its unique advantage is synergy. A framework for operationalizing and assessing partnership synergy, and for identifying its likely determinants, can be used to address critical policy, evaluation, and management issues related to collaboration. PMID- 11439465 TI - What happened to long-term care in the health reform debate of 1993-1994? Lessons for the future. AB - During 1993 and 1994, the United States debated but did not enact major health care reform. Although the reform efforts focused on providing health coverage for the uninsured and controlling acute care costs, many proposals included substantial long-term care initiatives. President Clinton proposed creating a large home-care program for severely disabled people of all ages and all income groups, among several other initiatives. By stressing non-means-tested public programs, the president's plan was a major departure from the Medicaid-dominated financing system for long-term care. In designing the long-term care component, the Clinton administration addressed many of the basic policy choices that must be decided in all reform efforts, including whether initiatives should be limited to older people or cover people of any age, how to balance institutional and noninstitutional care, whether to rely on government programs or on the private sector, and how to control costs. Analyzing the political and intellectual history of long-term care during the health reform debate provides lessons for future reform. PMID- 11439466 TI - Changes in the structure, composition, and activity of hospital governing boards, 1989-1997: evidence from two national surveys. AB - Hospital governance arrangements affect institutional policymaking and strategic decisions and can vary by such organizational attributes as ownership type/control, size, and system membership. A comparison of two national surveys shows how hospital governing boards changed in response to organizational and environmental pressures between 1989 and 1997. The magnitude and direction of changes in (1) board structure, composition, and selection; (2) CEO-board relations; and (3) board activity, evaluation, and compensation are examined for the population of hospitals and for different categories of hospitals. The findings suggest that hospital boards are engaging in selective rather than wholesale change to meet the simultaneous demands of a competitive market and traditional institutional orientations to community, the disenfranchised, and philanthropic service. Results also suggest parallel increases in collaboration between boards and CEOs and in board scrutiny of CEOs. PMID- 11439467 TI - Improving the quality of health care in the United Kingdom and the United States: a framework for change. AB - Fueled by public incidents and growing evidence of deficiencies in care, concern over the quality and outcomes of care has increased in both the United Kingdom and the United States. Both countries have launched a number of initiatives to deal with these issues. These initiatives are unlikely to achieve their objectives without explicit consideration of the multilevel approach to change that includes the individual, group/team, organization, and larger environment/system level. Attention must be given to issues of leadership, culture, team development, and information technology at all levels. A number of contingent factors influence these efforts in both countries, which must each balance a number of tradeoffs between centralization and decentralization in efforts to sustain the impetus for quality improvement over time. The multilevel change framework and associated properties provide a framework for assessing progress along the journey. PMID- 11439468 TI - Transport, environment and health. PMID- 11439469 TI - Attenuation and activation characteristics of steel and tungsten and the suitability of these materials for use in a fast neutron multileaf collimator. AB - A computer controlled multileaf collimator (MLC) is being designed to replace the multirod collimator (MRC) at present used to shape the d(48.5) + Be neutron beam from the Harper Hospital superconducting cyclotron. The computer controlled MLC will improve efficiency and allow for the future development of intensity modulated radiation therapy with neutrons. The existing MRC uses tungsten rods, while the new MLC will use steel as the leaf material. In the current study the attenuation and activation characteristics of steel are compared with those of tungsten to ensure that (a) the attenuation achieved in the MLC is at least equivalent to that of the existing MRC, and (b) that the activation of the steel will not result in a significant change in the activation levels within the treatment room. The latter point is important since personnel exposure (particularly to the radiation therapy technologists) from induced radioactivity must be minimized. Measurement of the neutron beam attenuation in a broad beam geometry showed that a 30 cm thick steel leaf yielded 2.5% transmission. This compared favorably with the 4% transmission obtained with the existing MRC. Irradiation of steel and tungsten samples at different depths in a 30 cm steel block indicated that the activation of steel should be no worse than that of tungsten. PMID- 11439470 TI - The water-equivalence of phantom materials for 90Sr-90Y beta particles. AB - Intravascular brachytherapy requires that the dose be specified within millimeters of the source. High dose gradients near brachytherapy sources require that the source-detector distance be accurately known for dosimetry purposes. Solid phantoms can be designed to accommodate these stringent requirements. This study reports dosimeter readings from 90Sr-90Y sources measured in water, A150, polystyrene and in an epoxy-based water-equivalent plastic. Measurements showed that while A150 and the epoxy-based plastic agreed well with water when the surface of the source contacted the detector housing, the relative response in the phantoms decreased with increasing depth in phantom, falling to approximately 0.55 those of water at a depth of 5 mm. Readings in polystyrene were within 4% of those in water between 1 and 2 mm depth. However, while polystyrene followed water more closely than the other two materials, at greater depths the relative response in polystyrene to water varied from 0.65 to 1.34. When the density of the materials is accounted for, the relative response in A150 is nearly constant with increasing areal density. Furthermore, the response in A150 shows the closest agreement with that in water of any of the solid materials for higher areal densities. For values below 0.3 g/cm2, polystyrene shows the closest agreement with water. PMID- 11439471 TI - Calculation of mean central dose in interstitial brachytherapy using Delaunay triangulation. AB - In 1997 the ICRU published Report 58 "Dose and Volume Specification for Reporting Interstitial Therapy" with the objective of addressing the problem of absorbed dose specification for reporting contemporary interstitial therapy. One of the concepts proposed in that report is "mean central dose." The fundamental goal of the mean central dose (MCD) calculation is to obtain a single, readily reportable and intercomparable value which is representative of dose in regions of the implant "where the dose gradient approximates a plateau." Delaunay triangulation (DT) is a method used in computational geometry to partition the space enclosed by the convex hull of a set of distinct points P into a set of nonoverlapping cells. In the three-dimensional case, each point of P becomes a vertex of a tetrahedron and the result of the DT is a set of tetrahedra. All treatment planning for interstitial brachytherapy inherently requires that the location of the radioactive sources, or dwell positions in the case of HDR, be known or digitized. These source locations may be regarded as a set of points representing the implanted volume. Delaunay triangulation of the source locations creates a set of tetrahedra without manual intervention. The geometric centers of these tetrahedra define a new set of points which lie "in between" the radioactive sources and which are distributed uniformly over the volume of the implant. The arithmetic mean of the dose at these centers is a three dimensional analog of the two-dimensional triangulation and inspection methods proposed for calculating MCD in ICRU 58. We demonstrate that DT can be successfully incorporated into a computerized treatment planning system and used to calculate the MCD. PMID- 11439472 TI - Validation of a two- to three-dimensional registration algorithm for aligning preoperative CT images and intraoperative fluoroscopy images. AB - We present a validation of an intensity based two- to three-dimensional image registration algorithm. The algorithm can register a CT volume to a single-plane fluoroscopy image. Four routinely acquired clinical data sets from patients who underwent endovascular treatment for an abdominal aortic aneurysm were used. Each data set was comprised of two intraoperative fluoroscopy images and a preoperative CT image. Regions of interest (ROI) were drawn around each vertebra in the CT and fluoroscopy images. Each CT image ROI was individually registered to the corresponding ROI in the fluoroscopy images. A cross validation approach was used to obtain a measure of registration consistency. Spinal movement between the preoperative and intraoperative scene was accounted for by using two fluoroscopy images. The consistency and robustness of the algorithm when using two similarity measures, pattern intensity and gradient difference, was investigated. Both similarity measures produced similar results. The consistency values were rotational errors below 0.74 degree and in-plane translational errors below 0.90 mm. These errors approximately relate to a two-dimensional projection error of 1.3 mm. The failure rate was less than 8.3% for three of the four data sets. However, for one of the data sets a much larger failure rate (28.5%) occurred. PMID- 11439473 TI - Advanced single-slice rebinning for tilted spiral cone-beam CT. AB - Future medical CT scanners and today's micro CT scanners demand cone-beam reconstruction algorithms that are capable of reconstructing data acquired from a tilted spiral trajectory where the vector of rotation is not necessarily parallel to the vector of table increment. For the medical CT scanner this case of nonparallel object motion is met for nonzero gantry tilt: the table moves into a direction that is not perpendicular to the plane of rotation. Since this is not a special application of medical CT but rather a daily routine in head exams, there is a strong need for corresponding reconstruction algorithms. In contrast to medical CT, where the special case of nonperpendicular motion is used on purpose, micro CT scanners cannot avoid aberrations of the rotational axis and the table increment vector due to alignment problems. Especially for those micro CT scanners that have the lifting stage mounted on the rotation table (in contrast to setups where the lifting stage holds the rotation table), this kind of misalignment is equivalent to a gantry tilt. We therefore generalize the advanced single-slice rebinning algorithm (ASSR), which is considered a very promising approach for medical cone-beam reconstruction due to its high image quality and its high reconstruction speed [Med. Phys. 27, 754-772 (2000)], to the case of tilted gantries. We evaluate this extended ASSR approach (which we will denote as ASSR+, for convenience) in comparison to the original ASSR algorithm using simulated phantom data for reconstruction. For the case of nonparallel object motion ASSR+ shows significant improvements over ASSR, however, its computational complexity is slightly increased due to the broken symmetry of the spiral trajectory. PMID- 11439474 TI - A cone beam filtered backprojection (CB-FBP) reconstruction algorithm for a circle-plus-two-arc orbit. AB - The circle-plus-arc orbit possesses advantages over other "circle-plus" orbits for the application of x-ray cone beam (CB) volume CT in image-guided interventional procedures requiring intraoperative imaging, in which movement of the patient table is to be avoided. A CB circle-plus-two-arc orbit satisfying the data sufficiency condition and a filtered backprojection (FBP) algorithm to reconstruct longitudinally unbounded objects is presented here. In the circle suborbit, the algorithm employs Feldkamp's formula and another FBP implementation. In the arc suborbits, an FBP solution is obtained originating from Grangeat's formula, and the reconstruction computation is significantly reduced using a window function to exclude redundancy in Radon domain. The performance of the algorithm has been thoroughly evaluated through computer simulated phantoms and preliminarily evaluated through experimental data, revealing that the algorithm can regionally reconstruct longitudinally unbounded objects exactly and efficiently, is insensitive to the variation of the angle sampling interval along the arc suborbits, and is robust over practical x-ray quantum noise. The algorithm's merits include: only 1D filtering is implemented even in a 3D reconstruction, only separable 2D interpolation is required to accomplish the CB backprojection, and the algorithm structure is appropriate for parallel computation. PMID- 11439475 TI - Computerized image analysis: estimation of breast density on mammograms. AB - An automated image analysis tool is being developed for the estimation of mammographic breast density. This tool may be useful for risk estimation or for monitoring breast density change in prevention or intervention programs. In this preliminary study, a data set of 4-view mammograms from 65 patients was used to evaluate our approach. Breast density analysis was performed on the digitized mammograms in three stages. First, the breast region was segmented from the surrounding background by an automated breast boundary-tracking algorithm. Second, an adaptive dynamic range compression technique was applied to the breast image to reduce the range of the gray level distribution in the low frequency background and to enhance the differences in the characteristic features of the gray level histogram for breasts of different densities. Third, rule-based classification was used to classify the breast images into four classes according to the characteristic features of their gray level histogram. For each image, a gray level threshold was automatically determined to segment the dense tissue from the breast region. The area of segmented dense tissue as a percentage of the breast area was then estimated. To evaluate the performance of the algorithm, the computer segmentation results were compared to manual segmentation with interactive thresholding by five radiologists. A "true" percent dense area for each mammogram was obtained by averaging the manually segmented areas of the radiologists. We found that the histograms of 6% (8 CC and 8 MLO views) of the breast regions were misclassified by the computer, resulting in poor segmentation of the dense region. For the images with correct classification, the correlation between the computer-estimated percent dense area and the "truth" was 0.94 and 0.91, respectively, for CC and MLO views, with a mean bias of less than 2%. The mean biases of the five radiologists' visual estimates for the same images ranged from 0.1% to 11%. The results demonstrate the feasibility of estimating mammographic breast density using computer vision techniques and its potential to improve the accuracy and reproducibility of breast density estimation in comparison with the subjective visual assessment by radiologists. PMID- 11439476 TI - Automated registration of breast lesions in temporal pairs of mammograms for interval change analysis--local affine transformation for improved localization. AB - Analysis of interval change is important for mammographic interpretation. The aim of this study is to evaluate the use of an automated registration technique for computer-aided interval change analysis in mammography. Previously we developed a regional registration technique for identifying masses on temporal pairs of mammograms. In the current study, we improved lesion registration by including a local alignment step. Initially, the lesion position on the prior mammogram was estimated based on the breast geometry. An initial fan-shaped search region was then defined on the prior mammogram. In the second stage, the location of the fan shaped region on the prior mammogram was refined by warping, based on an affine transformation and simplex optimization in a local region. In the third stage, a search for the best match between the lesion template from the current mammogram and a structure on the prior mammogram was carried out within the search region. This technique was evaluated on 124 temporal pairs of mammograms containing biopsyproven masses. Eighty-seven percent of the estimated lesion locations resulted in an area overlap of at least 50% with the true lesion locations and an average distance of 2.4 +/- 2.1 mm between their centroids. The average distance between the estimated and the true centroid of the lesions on the prior mammogram over all 124 temporal pairs was 4.2 +/- 5.7 mm. The registration accuracy was improved in comparison with our previous study that used a data set of 74 temporal pairs of mammograms. This improvement in accuracy resulted from the improved geometry estimation and the local affine transformation. PMID- 11439477 TI - Regional improvement of signal-to-noise and contrast-to-noise ratios in dual screen CR chest imaging--a phantom study. AB - The improvement of signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) in dual-screen computed radiography (CR) has been investigated for various regions in images of an anthropomorphic chest phantom. With the dual-screen CR technique, two image plates are placed in a cassette and exposed together during imaging. The exposed plates are separately scanned to form a front image and a back image, which are then registered and superimposed to form a composite image with improved SNRs and CNRs. The improvement can be optimized by applying specifically selected weighting factors during superimposition. In this study, dual-screen CR images of an anthropomorphic chest phantom were acquired and formed with four different combinations of standard resolution (ST) and high resolution (HR) screens: ST-ST, ST-HR, HR-ST, and HR-HR. SNRs and their improvements were measured and compared over twelve representative regions-of interest (ROIs) in these images. A 19.1%-45.7% increase of the SNR was observed, depending on the ROI and screen combination used. The optimal weighting factors were found to vary by only 4.5%-12.4%. Largest improvement was found in the lung field for all screen combinations. Improvement of CNRs was investigated over two ROIs in the lung field using the rib bones as the contrast objects and a 29.2% 43.9% improvement of the CNR was observed. Among the four screen combinations, ST ST resulted in the most SNR and CNR improvement, followed in order by HR-ST, HR HR, and ST-HR. The HR-ST combination yielded the lowest spatial variation of the optimal weighting factors with improved SNRs and CNRs close to those of the ST-ST combination. PMID- 11439478 TI - An automated patient recognition method based on an image-matching technique using previous chest radiographs in the picture archiving and communication system environment. AB - An automated patient recognition method for correcting "wrong" chest radiographs being stored in a picture archiving and communication system (PACS) environment has been developed. The method is based on an image-matching technique that uses previous chest radiographs. For identification of a "wrong" patient, the correlation value was determined for a previous image of a patient and a new, current image of the presumed corresponding patient. The current image was shifted horizontally and vertically and rotated, so that we could determine the best match between the two images. The results indicated that the correlation values between the current and previous images for the same, "correct" patients were generally greater than those for different, "wrong" patients. Although the two histograms for the same patient and for different patients overlapped at correlation values greater than 0.80, most parts of the histograms were separated. The correlation value was compared with a threshold value that was determined based on an analysis of the histograms of correlation values obtained for the same patient and for different patients. If the current image is considered potentially to belong to a "wrong" patient, then a warning sign with the probability for a "wrong" patient is provided to alert radiology personnel. Our results indicate that at least half of the "wrong" images in our database can be identified correctly with the method described in this study. The overall performance in terms of a receiver operating characteristic curve showed a high performance of the system. The results also indicate that some readings of "wrong" images for a given patient in the PACS environment can be prevented by use of the method we developed. Therefore an automated warning system for patient recognition would be useful in correcting "wrong" images being stored in the PACS environment. PMID- 11439479 TI - A semi-automatic technique for measurement of arterial wall from black blood MRI. AB - Black blood magnetic resonance imaging (MRI) has become a popular technique for imaging the artery wall in vivo. Its noninvasiveness and high resolution make it ideal for studying the progression of early atherosclerosis in normal volunteers or asymptomatic patients with mild disease. However, the operator variability inherent in the manual measurement of vessel wall area from MR images hinders the reliable detection of relatively small changes in the artery wall over time. In this paper we present a semi-automatic method for segmenting the inner and outer boundary of the artery wall, and evaluate its operator variability using analysis of variance (ANOVA). In our approach, a discrete dynamic contour is approximately initialized by an operator, deformed to the inner boundary, dilated, and then deformed to the outer boundary. A group of four operators performed repeated measurements on 12 images from normal human subjects using both our semiautomatic technique and a manual approach. Results from the ANOVA indicate that the inter operator standard error of measurement (SEM) of total wall area decreased from 3.254 mm2 (manual) to 1.293 mm2 (semi-automatic), and the intra-operator SEM decreased from 3.005 mm2 to 0.958 mm2. Operator reliability coefficients increased from less than 69% to more than 91% (inter-operator) and 95% (intra operator). The minimum detectable change in wall area improved from more than 8.32 mm2 (intra-operator, manual) to less than 3.59 mm2 (inter-operator, semi automatic), suggesting that it is better to have multiple operators measure wall area with our semi-automatic technique than to have a single operator make repeated measurements manually. Similar improvements in wall thickness and lumen radius measurements were also recorded. Since the semi-automatic technique has effectively ruled out the effect of the operator on these measurements, it may be possible to use such techniques to expand prospective studies of atherogenesis to multiple centers so as to increase access to real patient data without sacrificing reliability. PMID- 11439480 TI - Wavelength dependence of the precision of noninvasive optical measurement of oxy , deoxy-, and total-hemoglobin concentration. AB - The precision of noninvasive optical measurement of the concentration changes in oxy-, deoxy-, and total-hemoglobin depends on wavelength. For estimating the precision, we calculated the noise level of the concentration changes as the uncertainty in measurements using several wavelength pairs of light. Seven laser diodes (664-848 nm) were used simultaneously for spectroscopic measurement of brain activity during finger motor stimulation. We also used the analysis of error propagation from the uncertainty in direct measurements of absorbance changes to estimate indirectly the uncertainty of concentration changes. The measurement of the concentration changes made using an 830/664-nm pair are two times (oxy-Hb) and six times (deoxy-Hb) more precise than those made using an 830/782-nm pair. PMID- 11439481 TI - Accuracy limits in the determination of absolute optical properties using time resolved NIR spectroscopy. AB - We assess typical systematic experimental errors involved in a time-resolved measurement as applied to NIR diffuse optical spectroscopy and investigate their effect on the quantification accuracy of the absorption and the reduced scattering coefficient. We demonstrate that common systematic experimental uncertainties may lead to quantification errors of 10% or more, even when excellent signal to noise ratio conditions exist and accurate photon propagation models are employed. We further demonstrate that the accuracy of the calculation depends nonlinearly on the optical properties of the medium measured. High scattering and low absorbing media can be quantified more accurately than media with low scattering or high absorption using measurements of the same signal to noise ratio. We further discuss curve-shape fitting schemes that aid in improving the quantification accuracy in the presence of experimental errors. Finally, we identify uncertainties that set quantification accuracy limits and we find temporal resolution as the ultimate limiting factor in the quantification accuracy achieved. Our findings suggest that temporal resolution of the order of 10 ps is necessary for quantifying the absorption and reduced scattering coefficient of diffuse media with accuracy better than 5% using curve fitting methods. In that sense this analysis can be used in time-resolved system design and in predicting the expected errors given the technology selected for time resolved measurements. PMID- 11439482 TI - A model for the time dependent three-dimensional thermal distribution within iceballs surrounding multiple cryoprobes. AB - A time dependent three-dimensional finite difference model of iceball formation about multiple cryoprobes has been developed and compared to experimental data. Realistic three-dimensional probe geometry is specified and the number of cryoprobes, the cryoprobe cooling rates, and the locations of the probes are arbitrary inputs by the user. The simulation accounts for observed longitudinal thermal gradients along the cryoprobe tips. Thermal histories for several points around commercially available cryoprobes have been predicted within experimental error for one, three, and five probe configurations. The simulation can be used to generate isotherms within the iceball at arbitrary times. Volumes enclosed by the iceball and any isotherms may also be computed to give the ablative ratio, a measure of the iceball's killing efficiency. This ratio was calculated as the volume enclosed by a critical isotherm divided by the total volume of the iceball for assumed critical temperatures of -20 and -40 degrees C. The ablative ratio for a single probe is a continuously decreasing function of time but when multiple probe configurations are used the ablative ratio increases to a maximum and then essentially plateaus. Maximum values of 0.44 and 0.55 were observed for three and five probe configurations, respectively, with an assumed critical temperature of -20 degrees C. Assuming a critical temperature of -40 degrees C, maximum ablative ratios of 0.21 and 0.3 for three and five probe configurations, respectively, were observed. PMID- 11439483 TI - Comment on "Lens distortion in optically coupled digital x-ray imaging" [Med. Phys. 27, 906-912 (2000)]. PMID- 11439484 TI - The future will not need clinical therapy physicists. PMID- 11439485 TI - AAPM protocol for 40-300 kV x-ray beam dosimetry in radiotherapy and radiobiology. AB - The American Association of Physicists in Medicine (AAPM) presents a new protocol, developed by the Radiation Therapy Committee Task Group 61, for reference dosimetry of low- and medium-energy x rays for radiotherapy and radiobiology (40 kV < or = tube potential < or = 300 kV). It is based on ionization chambers calibrated in air in terms of air kerma. If the point of interest is at or close to the surface, one unified approach over the entire energy range shall be used to determine absorbed dose to water at the surface of a water phantom based on an in-air measurement (the "in-air" method). If the point of interest is at a depth, an in-water measurement at a depth of 2 cm shall be used for tube potentials > or = 100 kV (the "in-phantom" method). The in phantom method is not recommended for tube potentials < 100 kV. Guidelines are provided to determine the dose at other points in water and the dose at the surface of other biological materials of interest. The protocol is based on an up to-date data set of basic dosimetry parameters, which produce consistent dose values for the two methods recommended. Estimates of uncertainties on the final dose values are also presented. PMID- 11439486 TI - Leaf sequencing with secondary beam blocking under leaf positioning constraints for continuously modulated radiotherapy beams. AB - The creation of arbitrary photon fluence patterns for intensity modulated radiotherapy is addressed. The proposed method is intended for a class of multileaf collimators with a requirement for minimum leaf separation. Unlike the solution of Convery and Webb in which discrete beam intensity modulation was assumed, the present method deals with continuous modulation or that consisting of infinitely small bixels. The method begins with the time-optimal solution of Spirou-Stein-Svensson disregarding the minimum gap requirement. Subsequently, the gaps are restored by mobilizing the secondary beam blocking devices to prevent overexposure resulting from the leaf separation process. The secondary beam blocking is provided by means of two orthogonal backup diaphragms that are computer controlled. The results indicate that the method can be used to accurately deliver the desired modulation while satisfying the leaf positioning constraints. Furthermore, an example is presented which illustrates the efficacy of using the horizontal backup diaphragms (moving in perpendicular direction of the leaves) in addition to the vertical backup diaphragms (moving in the parallel direction of the leaves) to generate zero fluence regions. PMID- 11439487 TI - Acceleration of dose calculations for intensity-modulated radiotherapy. AB - The requirements and trade-offs between accuracy and speed for radiotherapy dose computations have been discussed for decades. Inverse planning used for intensity modulated radiotherapy (IMRT) optimization imposes additional demands on dose calculation since it is an iterative process in which dose calculations might be repeated many (10's to 1000's) of times. This work discusses the accuracy and speed issues as related to IMRT dose calculations. A hybrid dose calculation method which accelerates the optimization process is proposed and applied in which a fast-pencil beam (PB) model is used for initial optimization iterations, followed by superposition/convolution (SC) calculations. Optimization dose results are compared for pure PB optimization, pure SC optimization, and PB optimization followed by SC optimization. Plans were evaluated in terms of isodose coverage, dose-volume histograms, and total dose calculation time for five head and neck cases with diverse locations, sizes, and shapes for tumors and critical structures. Patient plans were designed for nine equispaced beams. For one patient, an additional five-beam configuration was tested. We found that gross features of intensity distributions resulting from all schemes were similar, however there were differences in the fine detail. Differences were small between composite dose distributions optimized with PB and SC methods, yet differences in individual beam dose distributions were quite significant. When the SC method was used to compute dose following optimization with PB method, dose differences were reduced significantly both for composite plans and for individual beams. Substantial overall timesavings were observed, allowing IMRT dose planning to become a more interactive activity. PMID- 11439488 TI - Dosimetric investigation and portal dose image prediction using an amorphous silicon electronic portal imaging device. AB - A two step algorithm to predict portal dose images in arbitrary detector systems has been developed recently. The current work provides a validation of this algorithm on a clinically available, amorphous silicon flat panel imager. The high-atomic number, indirect amorphous silicon detector incorporates a gadolinium oxysulfide phosphor scintillating screen to convert deposited radiation energy to optical photons which form the portal image. A water equivalent solid slab phantom and an anthropomorphic phantom were examined at beam energies of 6 and 18 MV and over a range of air gaps (approximately 20-50 cm). In the many examples presented here, portal dose images in the phosphor were predicted to within 5% in low-dose gradient regions, and to within 5 mm (isodose line shift) in high-dose gradient regions. Other basic dosimetric characteristics of the amorphous silicon detector were investigated, such as linearity with dose rate (+/- 0.5%), repeatability (+/- 2%), and response with variations in gantry rotation and source to detector distance. The latter investigation revealed a significant contribution to the image from optical photon spread in the phosphor layer of the detector. This phenomenon is generally known as "glare," and has been characterized and modeled here as a radially symmetric blurring kernel. This kernel is applied to the calculated dose images as a convolution, and is successfully demonstrated to account for the optical photon spread. This work demonstrates the flexibility and accuracy of the two step algorithm for a high atomic number detector. The algorithm may be applied to improve performance of dosimetric treatment verification applications, such as direct image comparison, backprojected patient dose calculation, and scatter correction in megavoltage computed tomography. The algorithm allows for dosimetric applications of the new, flat panel portal imager technology in the indirect configuration, taking advantage of a greater than tenfold increase in detector sensitivity over a direct configuration. PMID- 11439489 TI - Modeling the output ratio in air for megavoltage photon beams. AB - The output ratio in air, OR, for a high-energy x-ray beam describes how the incident central axis photon fluence varies with collimator setting. For field sizes larger than 3 x 3 cm2, its variation is caused by the scatter of photons in structures in the accelerator head (primarily the flattening filter and the wedge, if one is used) and by the backscatter of radiation into the monitor ionization chamber. The objective of this study was to evaluate the use of an analytical function to parametrize OR for square collimator setting c: OR = (1 + a1.c).[1 + a2.erf(c/lambda)2].H0. For open beams, these parameters can be attributed to explicit physical meanings within the systematical uncertainty of the model: a1 accounts for backscatter into the monitor, a2 is the maximum scatter-to-primary ratio for head-scattered photons, and lambda represents the effective width of the "source" of head-scatter photons. H0 is a constant that sets OR = 1 for c = 10 cm. This formula also fits OR for wedge beams and a Co-60 unit, although the fitting parameters lose their physical interpretations. To calculate the output ratio for a rectangular field, cx x cy, an equivalent square can be used: c = (1 + k).cy x cx/(k.cx + cy), where k is a constant. The study included a number of different accelerators and a cobalt-60 unit. The fits for square fields agreed with measurements with a standard deviation (SD) of less than 0.5%. Using k = lx.(f - ly)/ly.(f - lx), where lx and ly are the source-to collimator distances and f is the source-to-detector distance, measurements and calculations agreed within a SD of 0.7% for rectangular fields. Sufficient data for the three parameters are presented to suggest constraints that can be used for quality assurance of the measured output ratio in air. PMID- 11439490 TI - Monte Carlo calculation of output factors for circular, rectangular, and square fields of electron accelerators (6-20 MeV). AB - Monte Carlo (MC) techniques can be used to build a simulation model of an electron accelerator to calculate output factors for electron fields. This can be useful during commissioning of electron beams from a linac and in clinical practice where irregular fields are also encountered. The Monte Carlo code BEAM/EGS4 was used to model electron beams (6-20 MeV) from a Varian 2100C linear accelerator. After optimization of the Monte Carlo simulation model, agreement within 1% to 2% was obtained between calculated and measured (with a Si diode) lateral and depth dose distributions or within 1 mm in the penumbral regions. Output factors for square, rectangular, and circular fields were measured using two different plane-parallel ion chambers (Markus and NACP) and compared to MC simulations. The agreement was usually within 1% to 2%. This study was not primarily concerned with minimizing the simulation time required to obtain output factors but some considerations with respect to this are presented. It would be particularly useful if the MC model could also be used to calculate output factors for other, similar linacs. To see if this was possible, the primary electron energies in the MC model were retuned to model a recently commissioned similar linac. Good agreement between calculated and measured output factors was obtained for most field sizes for this second accelerator. PMID- 11439491 TI - A measured data set for evaluating electron-beam dose algorithms. AB - The purpose of this work was to develop an electron-beam dose algorithm verification data set of high precision and accuracy. Phantom geometries and treatment-beam configurations used in this study were similar to those in a subset of the verification data set produced by the Electron Collaborative Working Group (ECWG). Measurement techniques and quality-control measures were utilized in developing the data set to minimize systematic errors inherent in the ECWG data set. All measurements were made in water with p-type diode detectors and using a Wellhofer dosimetry system. The 9 and 20 MeV, 15 x 15 cm2 beams from a single linear accelerator composed the treatment beams. Measurements were made in water at 100 and 110 cm source-to-surface distances. Irregular surface measurements included a "stepped surface" and a "nose-shaped surface." Internal heterogeneity measurements were made for bone and air cavities in differing orientations. Confidence in the accuracy of the measured data set was reinforced by a comparison with Monte Carlo (MC)-calculated dose distributions. The MC calculated dose distributions were generated using the OMEGA/BEAM code to explicitly model the accelerator and phantom geometries of the measured data set. The precision of the measured data, estimated from multiple measurements, was better than 0.5% in regions of low-dose gradients. In general, the agreement between the measured data and the MC-calculated data was within 2%. The quality of the data set was superior to that of the ECWG data set, and should allow for a more accurate evaluation of an electron beam dose algorithm. The data set will be made publicly available from the Department of Radiation Physics at The University of Texas M. D. Anderson Cancer Center. PMID- 11439492 TI - Backscatter dose from metallic materials due to obliquely incident high-energy photon beams. AB - If metallic material is exposed to ionizing radiation of sufficient high energy, an increase in dose due to backscatter radiation occurs in front of this material. Our purpose in this study was to quantify these doses at variable distances between scattering materials and the detector at axial beam angles between 0 degree (zero angle in beams eye view) and 90 degrees. Copper, silver and lead sheets embedded in a phantom of perspex were exposed to 10 MV bremsstrahlung. The detector we developed is based on the fluorescence property of pyromellitic acid (1,2,4,5 benzenetetracarboxylic acid) after exposure to ionizing radiation. Our results show that the additional doses and the corresponding dose distribution in front of the scattering materials depend quantitatively and qualitatively on the beam angle. The backscatter dose increases with varying beam angle from 0 degree to 90 degrees up to a maximum at 55 degrees for copper and silver. At angles of 0 degree and 55 degrees the integral backscatter doses over a tissue-equivalent depth of 2 mm are 11.2% and 21.6% for copper and 24% and 28% for silver, respectively. In contrast, in front of lead there are no obvious differences of the measured backscatter doses at angles between 0 degree and 55 degrees. With a further increase of the beam angle from 55 degrees to 90 degrees the backscatter dose decreases steeply for all three materials. In front of copper a markedly lower penetrating depth of the backscattered electrons was found for an angle of 0 degree compared to 55 degrees. This dependence from the beam angle was less pronounced in front of silver and not detectable in front of lead. In conclusion, the dependence of the backscatter dose from the angle between axial beam and scattering material must be considered, as higher scattering doses have to be considered than previously expected. This may have a clinical impact since the surface of metallic implants is usually curved. PMID- 11439493 TI - Variation of sensitometric curves of radiographic films in high energy photon beams. AB - Film dosimetry is an important tool for the verification of irradiation techniques. The shape of the sensitometric curve depends on the type of film as well as on the irradiation and processing conditions. Existing data concerning the influence of irradiation geometry on the sensitometric curve are conflicting. In particular the variation of optical density, OD, with field size and depth in a phantom shows large differences in magnitude between various authors. This variation, as well as the effect of beam energy and film plane orientation on OD, was therefore investigated for two types of film, Kodak X-Omat V and Agfa Structurix D2. Films were positioned in a solid phantom, either perpendicular or (almost) parallel to the beam axis, and irradiated to different dose levels using various photon beams (Co-60, 6 MV, 15 MV, 18 MV, 45 MV). It was found that the sensitometric curves of the Kodak film derived at different depths are almost identical for the four x-ray beams. For the Kodak film the differences in OD with depth are less than 2%, except for the Co-60 beam, where the difference is about 4% at 10 cm depth for a 15 cm x 15 cm field. The slope of the sensitometric curve of the Agfa film is somewhat more dependent on photon beam energy, depth and field size. The sensitometric curves of both types of film are almost independent of the film plane orientation, except for shallow depths. For Co-60 and for the same dose, the Kodak and Agfa films gave at dose maximum an OD lower by 4% and 6%, respectively, for the parallel compared to the perpendicular geometry. Good dosimetric results can be obtained if films from the same batch are irradiated with small to moderate field sizes (up to about 15 cm x 15 cm), at moderate depths (up to about 15 cm), using a single calibration curve, e.g., for a 10 cm x 10 cm field. PMID- 11439494 TI - Experimental determination and verification of the parameters used in a proton pencil beam algorithm. AB - We present an experimental procedure for the determination and the verification under practical conditions of physical and computational parameters used in our proton pencil beam algorithm. The calculation of the dose delivered by a single pencil beam relies on a measured spread-out Bragg peak, and the description of its radial spread at depth features simple specific parameters accounting individually for the influence of the beam line as a whole, the beam energy modulation, the compensator, and the patient medium. For determining the experimental values of the physical parameters related to proton scattering, we utilized a simple relation between Gaussian radial spreads and the width of lateral penumbras. The contribution from the beam line has been extracted from lateral penumbra measurements in air: a linear variation with the distance collimator-point has been observed. Analytically predicted radial spreads within the patient were in good agreement with experimental values in water under various reference conditions. Results indicated no significant influence of the beam energy modulation. Using measurements in presence of Plexiglas slabs, a simple assumption on the effective source of scattering due to the compensator has been stated, leading to accurate radial spread calculations. Dose measurements in presence of complexly shaped compensators have been used to assess the performances of the algorithm supplied with the adequate physical parameters. One of these compensators has also been used, together with a reference configuration, for investigating a set of computational parameters decreasing the calculation time while maintaining a high level of accuracy. Faster dose computations have been performed for algorithm evaluation in the presence of geometrical and patient compensators, and have shown good agreement with the measured dose distributions. PMID- 11439495 TI - A theoretical model for event statistics in microdosimetry. I: Uniform distribution of heavy ion tracks. AB - In this work we describe a novel approach to solving microdosimetry problems using conditional probabilities and geometric concepts. The intersection of a convex site with a field of randomly oriented straight track segments is formulated in terms of the relative overlap between the chord associated with the action line of the track and the track itself. This results in a general formulation that predicts the contribution of crossers, stoppers, starters, and insiders in terms of two separate functions: the chord length distribution (characteristic of the site geometry and the type of randomness) and an independent set of conditional probabilities. A Monte Carlo code was written in order to validate the proposed approach. The code can represent the intersection between an isotropic field of charged particle tracks and a general ellipsoid of unrestricted geometry. This code was used to calculate the event distribution for a sphere as well as the expected mean value and variance of the track length distribution and to compare these against the deterministic calculations. The observed agreement was shown to be very good, within the precision of the Monte Carlo approach. The formulation is used to calculate the event frequency, lineal energy, and frequency mean specific energy for several monoenergetic and isotropic proton fields in a spherical site, as a function of the site diameter, proton energy, and the event type. PMID- 11439496 TI - A theoretical model for event statistics in microdosimetry. II: Nonuniform distribution of heavy ion tracks. AB - A microdosimetry model, described in Part I, applies to the case of a convex site immersed in a uniform distribution of heavy particle tracks, and assumes no restrictions in site geometry or the kind of randomness. In Part II, this model is extended to include nonuniform distributions of particle tracks. This situation is relevant to the study of microdosimetry, for example, in boron neutron capture, in irradiation experiments using heavy ion particle beams, where the sources of particle tracks are external to the cell, or in irradiation from internally incorporated particle-emitting radionuclides, such as environmental radon or occupational exposure to radioactive materials. The formalism developed permits the calculation of statistical properties, track length distributions, and microdosimetric spectra for convex sites where the "inner" and "outer" concentrations of sources may be different, or for tracks originating on the surface of a convex site. Expressions applicable to the case of surface distributed sources of tracks are presented that may represent situations such as boron compounds bound to the membrane of a cellular nucleus in boron neutron capture. A series of Monte Carlo calculations and analytical solutions, illustrating the case of spherical site geometry, are presented and compared. Finally, microdosimetric spectra and specific energy averages are calculated for alpha and lithium particles originating from thermal neutron capture in 10B, showing their dependence on 10B localization (extra-site, uniform, intra-site, or surface-distributed). PMID- 11439497 TI - What JCAHO surveyors are looking for: reports from an actual survey. PMID- 11439498 TI - Colling: what to look for when your hospital orders a security survey. PMID- 11439499 TI - A program aimed at improving security dept employee satisfaction. PMID- 11439501 TI - Meeting the major threats to hospital security within a budget. PMID- 11439500 TI - Infant kidnappings: new approaches; realistic drills; making low-tech and high tech improvements. AB - Although over the past few years there has been a decline in the number of infant kidnappings reported annually by hospitals, pressure on hospital officials by regulators, notably JCAHO, to do a better job of securing maternity areas has increased (see the article beginning on page one of this issue). And despite heavy investments in electronic detection systems, infant abductions continue to take place causing many hospital officials to take a second look at such systems (see the article beginning on page one of the January 2001 issue). In this report, we'll give details about a recent abduction, what happened, and what steps the hospital is taking to prevent a recurrence. We'll also relate how one hospital is devising and conducting regular infant abduction drills. And we'll fill you in on improvements that have been made in the widely used Code Alert system. PMID- 11439502 TI - [Nutritional assessment by anthropometric measurements]. PMID- 11439503 TI - [Nutritional assessment by bioelectrical impedance analysis]. PMID- 11439504 TI - [Functional nutritional assessment]. PMID- 11439505 TI - [Biochemical examination of blood and urine for nutritional assessment]. PMID- 11439506 TI - [Immunological parameters for nutritional assessment]. PMID- 11439507 TI - [Nutritional assessment by analysis of Rapid turnover proteins (RTP)]. PMID- 11439508 TI - [Nutritional assessment by Indirect calorimetry]. PMID- 11439509 TI - [Indices for nutritional assessment with multiple parameters]. PMID- 11439510 TI - [Energy and nutritional assessment and requirement]. PMID- 11439511 TI - [Indications for the nutritional support and route of administration]. PMID- 11439512 TI - [Assessment for effects of the nutritional support]. PMID- 11439513 TI - [Base solution containing dextrose (and others) and electrolytes (and amino acid) injection fluid for TPN]. PMID- 11439514 TI - [Amino acid solutions, and their features]. PMID- 11439515 TI - [Lipid emulsion]. PMID- 11439516 TI - [Multivitamin preparations for total parenteral nutrition]. PMID- 11439517 TI - [Current micro-element preparations for parenteral nutritional use]. PMID- 11439518 TI - [Characteristics, indication and contraindication of parenteral nutrition]. PMID- 11439519 TI - [Solutions for the hyperalimentation]. PMID- 11439520 TI - [Electrolyte solutions for total parenteral nutrition]. PMID- 11439521 TI - [TPN solution prepared by hospital pharmacy]. PMID- 11439522 TI - [Preparation of intravenous hyperalimentation solution]. PMID- 11439523 TI - [The kinds and characteristics of blood accesses]. PMID- 11439524 TI - [Know-how of catheter cannulation]. PMID- 11439525 TI - [Introduction, scheduling and rate of infusion for TPN in surgical patients]. PMID- 11439526 TI - [Control in TPN treatment--fundamentals]. PMID- 11439527 TI - [Careful management for avoiding an error or fluid therapy]. PMID- 11439528 TI - [Complications on the central vein catheterization]. PMID- 11439529 TI - [Enteral nutrition--characteristics, indication and contraindication]. PMID- 11439530 TI - [Catheter complications during parenteral nutrition]. PMID- 11439531 TI - [Diagnosis and treatment of central venous catheter sepsis]. PMID- 11439532 TI - [Early diagnosis and management of fungal infection in patients under parenteral nutirition]. PMID- 11439533 TI - [Hepatic complication of total parenteral nutrition]. PMID- 11439534 TI - [TPN induces bacterial translocation]. PMID- 11439535 TI - [Metabolic acidosis induced by TPN]. PMID- 11439536 TI - [Cholelithiasis in patients receiving long-term total parenteral nutrition]. PMID- 11439537 TI - [Other drugs administered with TPN solution]. PMID- 11439538 TI - [Peripheral parenteral nutrition]. PMID- 11439539 TI - [Properties of the available formulas for enteral nutrition]. PMID- 11439540 TI - [Oral supplement of BCAA in patients with liver disease]. PMID- 11439541 TI - [Beneficial effects of dietary fiber and oligosaccharide on enteral nutrition]. PMID- 11439542 TI - [Recent aspects and perspective of parenteral nutrition]. PMID- 11439543 TI - [Nutritional support team (NST)]. PMID- 11439544 TI - [Preparation of enteral feeds and bacterial contamination]. PMID- 11439545 TI - [Routes of enteral feeding and guidelines for applications in enteral nutrition]. PMID- 11439546 TI - [Nasogastric feeding tubes for enteral nutrition]. PMID- 11439547 TI - [Gastrostomy tube]. PMID- 11439548 TI - [Enteral feeding tubes]. PMID- 11439549 TI - [Enteral diet administration systems]. PMID- 11439550 TI - [Feeding pumps for enteral nutrition]. PMID- 11439551 TI - [Standard technique for the placement of enteral feeding tube]. PMID- 11439552 TI - [Plan to start enteral feeding, administration schedule and flow rate]. PMID- 11439553 TI - [General aspect on the management of enteral tube feeding]. PMID- 11439554 TI - [Mechanical complications associated with the tube for enteral nutrition]. PMID- 11439555 TI - [Abdominal complications during enteral nutrition]. PMID- 11439556 TI - [Combined nutritional management of total parenteral and enteral nutrition]. PMID- 11439557 TI - [Enteral nutrition and intestinal immunity]. PMID- 11439558 TI - [Drug administration for enteral nutrition]. PMID- 11439559 TI - [Outline of metabolic management and complications in parenteral and enteral nutrition]. PMID- 11439560 TI - [Glucose metabolism-metabolic complications and their managements]. PMID- 11439561 TI - [Metabolic management and complication of amino acid and protein metabolism]. PMID- 11439562 TI - [Metabolism of calcium, phosphorus, and bone]. PMID- 11439563 TI - [Lipid metabolism]. PMID- 11439564 TI - [Critical path in intravenous nutrition and tube feeding]. PMID- 11439565 TI - [Metabolism of trace elements during parenteral and enteral nutrition]. PMID- 11439566 TI - [Vitamin metabolism in parenteral and enteral nutrition]. PMID- 11439567 TI - [Intravenous and enteral nutrition for the patient with cerebrovascular disease]. PMID- 11439568 TI - [Total parenteral nutrition and forced enteral feeding for the patient with disturbance of consciousness]. PMID- 11439569 TI - [Nutritional management for the patient with neurosurgical disease]. PMID- 11439570 TI - [Nutritional support in cardiac failure]. PMID- 11439571 TI - [Nutritional support in cardiac cachexia]. PMID- 11439572 TI - [Management of nutrition in cardio-surgical patients]. PMID- 11439573 TI - [Parenteral and enteral nutrition in respiratory failure]. PMID- 11439574 TI - [Guidelines for the use of parenteral and enteral nutrition]. PMID- 11439575 TI - [Nutrition management in lung surgery]. PMID- 11439576 TI - [Nutritional support for ARDS]. PMID- 11439577 TI - [Parenteral and enteral nutrition for esophagitis and esophageal ulcer]. PMID- 11439578 TI - [Parenteral and enteral nutrition for esophageal cancer patient]. PMID- 11439579 TI - [Nutrition support for advanced esophageal cancer patients]. PMID- 11439580 TI - [Sclerotherapy and nutritional support for esophageal varices]. PMID- 11439581 TI - [Pre- and postoperative nutritional management for peptic ulcer patients]. PMID- 11439582 TI - [Parenteral and enteral nutritional management for hemorrhagic gastric and duodenal ulcer patients]. PMID- 11439583 TI - [The purpose and problem of total parenteral nutrition and tube feeding]. PMID- 11439584 TI - [Pre- and postoperative nutritional management for gastric cancer patients]. PMID- 11439585 TI - [Parenteral and enteral nutrition in the inoperable gastric cancer]. PMID- 11439586 TI - [Parenteral and enteral nutrition for surgical treatment of Crohn's disease]. PMID- 11439587 TI - [Parenteral and enteral nutrition for conservative treatment of Crohn's disease]. PMID- 11439588 TI - [Severity and parenteral enteral nutrition for ulcerative colitis]. PMID- 11439589 TI - [Nutritional management in patients receiving operation for intestinal Behcet's disease]. PMID- 11439590 TI - [Therapy and nutrition for intestinal Behcet disease and simple ulcer]. PMID- 11439591 TI - [Nutritional support for non-specific multiple ulcers of the small intestine]. PMID- 11439592 TI - [Nutritional support for protein losing gastroenteropathy]. PMID- 11439593 TI - [Nutritional care for the infants with necrotizing enterocolitis under non surgical treatment]. PMID- 11439594 TI - [An outline of the 6th revision Japanese dietary recommended allowance (dietary reference intake)]. PMID- 11439595 TI - [Neonatal necrotizing enterocolitis: nutritional care for surgical patients]. PMID- 11439596 TI - [Parenteral and enteral nutrition in surgical patients with intestinal obstruction]. PMID- 11439597 TI - [Parenteral and enteral nutrition in non-operative patients with intestinal obstruction]. PMID- 11439598 TI - [Total parenteral nutrition for anastomotic leakage and gastrointestinal fistulas]. PMID- 11439599 TI - [Perioperative and nutritional management for short bowel syndrome]. PMID- 11439600 TI - [Nutritional management of patients with malabsorption syndrome]. PMID- 11439601 TI - [Perioperative nutritional management for acute peritonitis patients]. PMID- 11439602 TI - [Perioperative parenteral and enteral nutritional management for colorectal cancer patients]. PMID- 11439603 TI - [Parenteral and enteral nutrition in far-advanced colorectal cancer]. PMID- 11439604 TI - [Nutritional management after small bowel transplantation]. PMID- 11439605 TI - [Total parenteral and enteral nutrition in hepatitis and hepatic failure]. PMID- 11439606 TI - [Pre and post operative nutritional management for the patients with hepatoma]. PMID- 11439607 TI - [Nutritional management in non surgical therapy of liver cancer]. PMID- 11439608 TI - [Fluid and nutritional management in living-related liver transplantation]. PMID- 11439609 TI - [Parenteral and enteral nutrition on cholecystitis and cholelithiasis]. PMID- 11439610 TI - [Nutritional support for surgery of the biliary tract cancer]. PMID- 11439611 TI - [Nutritional management for non-operative patients with biliary tract cancer]. PMID- 11439612 TI - [Perioperative nutritional management for patients with obstructive jaundice caused by biliary tract disease]. PMID- 11439613 TI - [Nutritional management for non operative patients with obstructive jaundice caused by biliary tract disease]. PMID- 11439614 TI - [Practical management of parenteral and enteral nutrition--from the standpoint of nursing team]. PMID- 11439615 TI - [Pre- and postoperative nutritional managements of acute pancreatitis]. PMID- 11439616 TI - [Fluids and nutrition in acute pancreatitis]. PMID- 11439617 TI - [Fluid and nutritional management in chronic pancreatitis before and after surgery]. PMID- 11439618 TI - [Nutritional care of chronic pancreatitis (pancreatolithiasis)]. PMID- 11439619 TI - [Metabolic and nutritional management for treatment of carcinoma of the pancreas- in resectable cases]. PMID- 11439620 TI - [Metabolic and nutritional management for treatment of carcinoma of the pancreas- in unresectable cases]. PMID- 11439621 TI - [Nutritional management on the cystic fibrosis]. PMID- 11439622 TI - [Parenteral and enteral nutrition in acute renal failure]. PMID- 11439623 TI - [Parenteral and enteral nutrition in chronic renal failure]. PMID- 11439624 TI - [Nutritional support for diabetes mellitus]. PMID- 11439625 TI - [Preoperative and postoperative nutritional management for diabetic patients]. PMID- 11439626 TI - [Parenteral and enteral nutrition on diabetic nephropathy]. PMID- 11439627 TI - [Parenteral and enteral nutrition in severe infection]. PMID- 11439628 TI - [Parenteral and enteral nutrition on HIV infection]. PMID- 11439629 TI - [Parenteral nutrition in entero-hemorrhagic E. coli infection]. PMID- 11439630 TI - [Parenteral and enteral nutrition for multiple organ failure]. PMID- 11439631 TI - [Parenteral nutrition on depleted patients]. PMID- 11439632 TI - [Enteral nutrition research--recent advances and future views]. PMID- 11439633 TI - [Parenteral and enteral nutrition for trauma patients]. PMID- 11439634 TI - [Enteral and parenteral nutrition in burns]. PMID- 11439635 TI - [Parenteral nutrition in obstetrics and gynecology]. PMID- 11439636 TI - [Parenteral and enteral nutrition in otorhinolaryngology]. PMID- 11439637 TI - [Total parenteral nutrition in urology]. PMID- 11439638 TI - [Intravenous and enteral nutrition in the field of dermatology (except burn)]. PMID- 11439639 TI - [Perioperative nutritional support in oral and maxillofacial surgery]. PMID- 11439640 TI - [Parenteral and enteral nutrition in plastic surgical practice]. PMID- 11439641 TI - [Parenteral nutrition on the cancer chemotherapy]. PMID- 11439642 TI - [Perioperative nutritional support]. PMID- 11439643 TI - [Parenteral and enteral fluid therapy for vomiting, diarrhea and dehydration]. PMID- 11439644 TI - [Recommended dietary allowances and nutritional assessments in infants and children]. PMID- 11439645 TI - [Specialties in parenteral and enteral nutritions in infants and children]. PMID- 11439646 TI - [Essential equipments for parenteral and enteral nutrition in pediatric patients]. PMID- 11439647 TI - [The route for nutrition in paediatric patients]. PMID- 11439648 TI - [Commercially available nutrients for parenteral and enteral feeding in infants and children]. PMID- 11439649 TI - [Present of parenteral and enteral management in dietitian]. PMID- 11439650 TI - [Indication and non-indication of parenteral and enteral nutritions in infants and children]. PMID- 11439651 TI - [Parenteral and enteral nutrition for premature infants]. PMID- 11439652 TI - [Parenteral and enteral nutrition for severe and protracted diarrhea in infancy]. PMID- 11439653 TI - [Parenteral and enteral nutrition in children with gastrointestinal diseases]. PMID- 11439654 TI - [Nutritional support in children with hepatobiliary diseases]. PMID- 11439655 TI - [Parenteral and enteral nutrition for children after open heart surgery]. PMID- 11439656 TI - [Nutritional support in children with cancer]. PMID- 11439657 TI - [Total parenteral nutrition in pediatric stem cell transplantation]. PMID- 11439658 TI - [Tube feeding for children with severe motor and intellectual disabilities]. PMID- 11439659 TI - [Parenteral and enteral nutrition in surgical patients in childhood]. PMID- 11439660 TI - [Nutritional assessment in the elderly]. PMID- 11439661 TI - [Nutritional support and fluid therapy in the elderly patients]. PMID- 11439662 TI - [Perioperative nutritional support for the aged]. PMID- 11439663 TI - [The practice of the parenteral and enteral nutrition--from the viewpoint of the pharmacist]. PMID- 11439664 TI - [Indication of home parenteral nutrition]. PMID- 11439665 TI - [Home parenteral nutrition for pediatric patients]. PMID- 11439666 TI - [Home parenteral nutrition in adults]. PMID- 11439667 TI - [Complication of HPN]. PMID- 11439668 TI - [Indication of home enteral hyperalimentation]. PMID- 11439669 TI - [Home enteral nutrition in pediatric surgical patients]. PMID- 11439670 TI - [Home enteral nutrition in adult patients]. PMID- 11439671 TI - [Problems of home enteral nutrition in aged]. PMID- 11439672 TI - [Complications and outcome of patients receiving home enteral nutrition]. PMID- 11439673 TI - [Home parenteral nutrition in cancer patients]. PMID- 11439674 TI - [Home nutrition therapy for Crohn's disease]. PMID- 11439675 TI - [The role of pharmacy in HIT (home infusion therapy)]. PMID- 11439676 TI - [Anti-cancer therapy with amino acid imbalance]. PMID- 11439677 TI - [Nutritional aspects of hGH or IGF-I supplementation in highly stressed patients]. PMID- 11439678 TI - [Multivitamin-prefiled-syringe as additives to the TPN]. PMID- 11439679 TI - [New total parenteral nutrition solution (GA-1080)]. PMID- 11439680 TI - [Glutamine]. PMID- 11439681 TI - [omega-3 Polyunsaturated fatty acid fat emulsion]. PMID- 11439682 TI - [Lipid emulsion containing MCFA, LCFA and omega-3 fatty acids]. PMID- 11439683 TI - [Nucleoside/nucleotide mixture]. PMID- 11439684 TI - [New polymeric liquid diet, OSN-001]. PMID- 11439685 TI - [Enteral nutrition--arginine, RNA, omega-3 fatty acids]. PMID- 11439686 TI - [New enteral nutrition products in the U.S. and Europe]. PMID- 11439687 TI - [Current topics of nutrition--new goods and drugs]. PMID- 11439688 TI - [Nutrition assessment]. PMID- 11439689 TI - Prescription for heart attack survivors. PMID- 11439690 TI - Falls and fractures: a preventive guide. PMID- 11439691 TI - Should you be taking tamoxifen? PMID- 11439692 TI - Foiling the ticks that make us sick. PMID- 11439693 TI - Are there any health reasons to stop drinking coffee? PMID- 11439694 TI - Can rosacea affect the eyes? PMID- 11439695 TI - Novel functional assay for proteases and modulators. Application in beta secretase studies. AB - Proteases play a critical role in many cellular functions and have been an attractive therapeutic target due to their involvement in a number of disease processes. One prominent example is the secretases responsible for the generation of amyloid beta peptide, which is believed to be central for the development of Alzheimer's disease. It is therefore desirable to identify and characterize these proteases. We have developed a novel functional approach for identification of proteases and modulators by coupling the protease activity to caspase-mediated apoptosis. Here we show the proof of principle for this approach using beta secretase as an example. We provide data showing that 1. A modified caspase-3 containing beta-secretase cleavage site induces apoptosis in 293T cells. 2. The modified caspase-3 induced apoptosis is correlated with the susceptibility of beta-secretase recognition sequence to beta-secretase. 3. In vivo beta-secretase competitors BACE2 and BACE2(D110A) prevent the modified caspase-3 induced cell death. Therefore, this approach can be a useful tool in studies of proteolytic cleavage provided only that the protease recognition sequence is known. PMID- 11439696 TI - Insights into T-cell development from studies using transgenic and knockout mice. AB - The generation of immunocompetent lymphocytes is a complex process that utilizes a multitude of cell surface receptors and intracellular signaling pathways. Moreover, specific cell-cell interactions and specialized microenvironments are required, so that purely in vitro experimental systems are limited in their ability to explain the complexity of T-cell development. In vivo models have been used extensively in the study of T-cell development. In the present review we summarize but a few of the seminal discoveries that have been made in this field using transgenic and knockout mouse models. In addition to demonstrating the wealth of information that can be gained, we also discuss some of the present limitations of this technology. Novel advances that allow the conditional and inducible modification of the genome and knock-in mutations promise to lead to an even more rapid advancement in our knowledge of T-cell development. PMID- 11439697 TI - Biolistic transformation of arbuscular mycorrhizal fungi. Progress and perspectives. AB - Gene transfer systems have proved effective for the transformation of a range of organisms for both fundamental and applied studies. Biolistic transformation is a powerful method for the gene transfer into various organisms and tissues that have proved recalcitrant to more conventional means. For fungi, the biolistic approach is particularly effective where protoplasts are difficult to obtain and/or the organisms are difficult to culture. This is particularly applicable to arbuscular mycorrhizal (AM) fungi, being as they are obligate symbionts that can only be propagated in association with intact plants or root explants. Furthermore, these fungi are aseptate and protoplasts cannot be released. Recent advancements in gene transformation systems have enabled the use of biolistic technology to introduce foreign DNA linked to molecular markers into these fungi. In this review we discuss the development of transformation strategies for AM fungi by biolistics and highlight the areas of this technology which require further development for the stable transformation of these elusive organisms. PMID- 11439698 TI - Oligonucleotide treatment of ras-induced tumors in nude mice. AB - Oligonucleotides have shown an ability to target specific oncogene transcripts and inhibit their expression in cells, but the degree to which sustained treatment can suppress the levels of an oncogenic protein enough to benefit a patient remains to be determined. This question has been studied in several ways. First, the relationship of antisense DNA inhibition to the predicted secondary structure of human H-RAS oncogene mRNA was examined in transformed mouse cells that form solid tumors. Inhibition of H-Ras expression was sequence-specific, dose-dependent, and correlated with inhibition of focus formation. The efficacy of the first intron antisense sequence in reducing H-Ras expression was greater than that of the initiation codon target. Second, H-RAS transformed solid tumor cells were pretreated in vitro with normal oligonucleotides, after which tumor growth from the treated cells was tested in nude mice. The three days of treatment with the first intron antisense DNA reduced H-Ras cellular levels by more than 90% whereas a nonspecific control DNA reduced H-Ras levels by approx 20%. Tumor growth of cells treated with H-RAS antisense oligonucleotide was significantly reduced for up to 14 d following the end of treatment and implantation into the mice, whereas the nonspecific control DNA had no significant effect. Third, H-RAS transformed bladder cancer cells were implanted into nude mice, after which the mice were treated for 31 d with oligonucleotide phosphorothioates. Tumor growth in mice treated with H-RAS 12th codon antisense oligonucleotide was reduced by about 80% throughout the treatment period, reiterating the sustained effect seen in pretreated tumor cells. However, the scrambled phosphorothioate control inhibited tumor growth by about 60%, illustrating some nonspecific inhibition. Fourth, K-RAS transformed pancreatic cancer cells were treated in culture and in nude mice. Inhibition of K-Ras expression with a phosphorothioate oligonucleotide directed against a 5'-UTR sequence was sequence-specific and dose-dependent. K-RAS transformed pancreatic cancer cells were implanted into nude mice, after which the mice were treated for 14 d with oligonucleotide phosphorothioates. Tumor growth in mice treated with K RAS 5'-UTR antisense oligonucleotide was reduced by about 50% throughout the treatment period, reiterating the sustained effect seen with H-RAS transformed cells. In this case, the sense phosphorothioate control did not inhibit tumor growth, demonstrating that nonspecific inhibition is not a characteristic of all phosphorothioate sequences. The next logical steps include testing oligonucleotide efficacy against other tumor types, toxicological testing in higher species, and clinical trials in human subjects. PMID- 11439699 TI - Cell adhesion assays. AB - Cell adhesion makes an important contribution to the maintenance of tissue structure, the promotion of cell migration, and the transduction of information about the cell microenvironment across the plasma membrane. An ability to quantitate adhesion has proven to be extremely valuable for those researchers studying the molecular mechanisms underlying these processes. This article will outline in detail two standard assays used for quantitating the adhesion of cells to an immobilized substrate. First, an attachment assay, which employs a colorimetric detection of bound cells, and second, a spreading assay, which employs phase contrast microscopy to measure the flattening of adherent cells. PMID- 11439700 TI - Construction of adenoviral vectors. AB - Recombinant adenovirus vectors have proven to be useful tools in facilitating gene transfer. Construction of such vectors requires a knowledge of the adenovirus genome structure and its life cycle. A commonly used recombinant adenovirus involves deletion of the E1 region; such a recombinant is traditionally produced by overlap recombination after cotransfection of 293 cells with a plasmid shuttle vector and a large right-end restriction fragment of viral DNA. The shuttle vector contains a cassette for a transgene placed in region E1 and flanking sequences from adenovirus for recombination. Normally, a high background of parental virus results because of the difficulty in separating right-end restriction fragment length DNA from uncut DNA. This paper describes a negative selection based on the traditional cotransfection method using viral DNA from an E1-deleted adenoviral recombinant that expresses green fluorescent protein (GFP). In situ fluorescent microscopy is used to distinguish the recombinant plaques (white or nonfluorescent) from the parental virus plaques (green or fluorescent). In addition, this system allows for the detection of contaminating parental virus at later stages when production lots of the recombinant vector are being made. PMID- 11439701 TI - Isolation and purification of chemokines from natural sources. AB - Chemokines (e.g., IL-8) were originally identified as chemotactic proteins obtained from various different natural sources. Today, using the genome walking strategy an ever-increasing number of novel genes encoding chemokines have been discovered that were expressed in bacteria or eukaryotic cells and subsequently tested for biological activity. Usually biological significance of the considered chemokine is extrapolated from these data. The increasing evidence, however, that post-translational modification of chemokines can dramatically affect its biological activity makes it necessary to identify the naturally occurring chemokines in order to identify its biological function. Furthermore, with the isolation of natural chemokines, evidence is provided that transcription of chemokine genes is really followed by translation into a bioactive molecule. Purification of chemokines from natural sources requires special strategies: The bioassay or immunoassay should allow screening of high-performance liquid chromatography fractions and detection of the required chemokine at low concentration. Parameters that affect the detection of bioactivity and immunoreactivity (giving either false positive or false negative results) should be carefully considered. In this article methods for molecular characterization of chemokines from both cell culture supernatants and human tissue (lesional inflammatory skin scales) will be described. PMID- 11439702 TI - Eligibility for the Medicare buy-in programs, based on a survey of income and program participation simulation. AB - Medicare buy-in programs are designed to reduce out-of-pocket expenses of beneficiaries with modest income and assets. This article provides estimates of the size of the Medicare beneficiary population eligible for the Qualified Medicare Beneficiary (QMB) program, the Specified Low-Income Medicare Beneficiary (SLMB) program, and the Qualified Individual-1 (QI-1) program. The buy-in programs use the same resource limits (twice those used in the Supplemental Security Income (SSI) program) but different thresholds for determining income eligibility. The QMB program uses 100 percent of the poverty line as the cutoff, QI-1 covers persons above 120 percent but at or below 135 percent of the poverty line, and the SLMB program is in between. Making informed judgments about the rate of participation in the buy-in programs and the need for outreach requires an accurate estimate of the size of the eligible population. If that population is underestimated, policymakers might come to unduly optimistic conclusions about current buy-in participation. In contrast, an overestimate may make current participation seem too low. If policymakers react to an upwardly biased estimate of the eligible population by increasing outreach, they are bound to be disappointed by the results of that effort. Estimates of the eligible population from past studies of the QMB and SLMB programs range from 5.1 million to 9.1 million. In the absence of new information, it is difficult to judge the accuracy of those estimates because the methodologies had substantial shortcomings that might bias the results. The most common shortcomings include the lack of high quality, monthly income data and the lack of information on assets from the same data file that was used to estimate participation and income eligibility for Medicare. The current study uses the most recently available (as of August 2000) Survey of Income and Program Participation (SIPP) file that is matched to the Social Security Administration's (SSA's) administrative records. The data file covers 1995 information. Estimates were also obtained using 1991 data to assess the sensitivity of eligibility estimates to the year chosen. The SIPP has several major advantages over other data sources because it contains relevant, high quality information on both income and assets for establishing financial eligibility for the buy-in programs. First, the SIPP collects detailed and conceptually appropriate information on monthly, rather than annual, income and therefore has more complete information about income than do other surveys. As a result, SIPP-based estimates of poverty are substantially lower than estimates based on the Current Population Survey. Second, the SIPP also collects information on assets at the individual level. Thus, the survey provides enough detail to measure the major income and asset exclusions directly. Finally, the SIPP data are matched to SSA administrative records: Medicare eligibility can therefore be accurately measured, and self-reported data on Social Security and SSI benefits can be replaced with more accurate monthly information. Our 1995 simulation estimates that approximately 4.8 million persons in the U.S. noninstitutionalized population were eligible for the QMB program and an additional 1.6 million for the SLMB program. The total--roughly 6.5 million--is within the range of estimates from past studies but is closer to the lower end, suggesting that the eligible population is smaller than was previously believed. When the estimated QI-1 eligible population of 0.9 million is added, the total for the three buy-in programs is 7.4 million. Because the QI-1 program did not exist in 1995, only the estimated 6.5 million QMBs and SLMBs would actually have been eligible to receive benefits. The 7.4 million figure represents the 1995 Medicare beneficiaries who would be eligible for buy-in under program rules for 2000. Adjusting that number to account for increases in the Medicare population between 1995 and 1999 yields an estimated eligible population of 7.8 million in 1999. Compared with other elderly Medicare recipients, eligible elderly QMBs and SLMBs have poorer health, more functional limitations, and higher rates of health care use. Thus, not only are their income resources relatively limited, but their need for potentially expensive medical care is also greater. Similar differences were not found in health, functional limitations, and health care use among disabled participants in the QMB and SLMB programs. Our estimates imply that about 2.5 million noninstitutionalized individuals were eligible for but not enrolled in the QMB and SLMB programs in 1999. That finding suggests that fewer eligibles may be available for targeting by outreach efforts than was previously believed. Outreach may be more difficult than it would be with a larger eligible population. (ABSTRACT TRUNCATED) PMID- 11439703 TI - Medicare premium buy-in programs: results of SSA demonstration projects. AB - Three programs known collectively as the Medicare buy-in programs are available to pay Medicare Part B premiums and, in some cases, other medical expenses for certain low-income individuals. The Health Care Financing Administration administers those programs, with most functions performed by the states. The Social Security Administration (SSA) plays an indirect role in the buy-in programs: with certain exceptions, people who qualify for Medicare and hence for buy-in are beneficiaries of Social Security retirement or disability programs. SSA is often cited as an agency that might be able to increase enrollment in the buy-in programs through outreach to its beneficiaries and by acting as an intermediary in the enrollment process. The three buy-in programs have different requirements for eligibility. The Qualified Medicare Beneficiary (QMB) program includes individuals who have Part A Medicare benefits and whose income does not exceed 100 percent of federal poverty guidelines. People in the Specified Low Income Medicare Beneficiary (SLMB) program are individuals who would otherwise be QMBs but whose income is more than 100 percent but less than 120 percent of poverty guidelines. People in the Qualified Individual (QI) program are those who meet the other criteria but whose income is less than 175 percent of poverty guidelines. Various reports and studies by government agencies and advocacy organizations conclude that the buy-in programs are not reaching many of the people who are eligible. Low enrollment appears to be a particular issue for the SLMB and QI programs. States have tried various outreach efforts, but the effectiveness of those efforts has not been adequately assessed. In 1998, Congress mandated that SSA conduct a demonstration project to determine how to increase participation in the buy-in programs. The project tested six different administrative models in which outreach letters were sent to potential beneficiaries asking them to contact SSA and then be screened for eligibility and referred for enrollment. SSA was able to screen about 7.1 percent of letter recipients for buy-in eligibility: 4.2 percent were potentially eligible for the programs based on income and resources, and 3.7 percent enrolled in a buy-in program. An evaluation of the probability that letter recipients would contact SSA to be screened found that: Among the elderly, older individuals were less likely to be screened but more likely to enroll. Among the disabled, older individuals were more likely to be screened but less likely to enroll. The disabled were less likely to be screened but more likely to enroll. Individuals with higher Social Security benefits were more likely to be screened but less likely to enroll. Women were more likely to be screened and to enroll. Being married did not appear to affect screening but negatively affected enrollment. Individuals with a preference for materials in Spanish were much more likely to be screened and enrolled. In some of the demonstration sites, enrollment in a Medicare+Choice plan increased the probability of being both screened and enrolled. SSA conducted a survey of some people who did not respond to the outreach letter. Most of those from whom explanations of the nonresponse were obtained had not responded because they were not eligible on the basis of their income or resources. If SSA were to reproduce the demonstrations in a nationwide outreach effort, a national mailing would include nearly 20 million individuals. If response rates were similar to those seen in the 1999 demonstrations, outreach would produce over 740,000 new buy-in enrollees. That number might be increased modestly by conducting additional outreach efforts in conjunction with the mailing. PMID- 11439704 TI - Divorced women at retirement: projections of economic well-being in the near future. AB - The Modeling Income in the Near Term (MINT) data system projects retirement income for persons retiring in the 1990s through 2020. Using those data, we examine the economic well-being of divorced women at retirement. The MINT data system improves upon previous estimates of Social Security benefits by: Measuring and projecting years of marriage to determine if the 10-year requirement has been met, Projecting lifetime earnings until retirement and eligibility for Social Security retirement benefits, and Estimating lifetime earnings of former spouses. MINT also makes independent projections of each retiree's income from pensions, assets, and earnings (for working beneficiaries). As a result of changes in marital patterns, MINT projects that the proportion of women who are divorced will increase. At the same time, the proportion of those women who are eligible for auxiliary benefits is projected to decrease, for two main reasons. First, changes in women's earnings and work patterns result in more women receiving retired-worker benefits based on their own earnings. Second, an increased number of divorced women will not meet the 10-year marriage requirement for auxiliary benefits. Despite the projected decrease over time in eligibility rates for auxiliary benefits, the level of Social Security benefits is projected to change little between the older and younger birth cohorts of divorced women entering retirement. According to the MINT data, the most vulnerable of divorced women will be those who have not met the 10-year marriage requirement. Poverty rates will be higher for them than for all other divorced women. This group of divorced women is projected to grow as more and more women divorce from shorter marriages. With more women divorcing and with fewer divorced women meeting the 10-year marriage requirement, the proportion of economically vulnerable aged women will increase when the baby boom retires. Further research is warranted on this long neglected subject. Analyses of divorced women's economic well-being by major socioeconomic characteristics such as race and ethnicity and education are of particular interest. Such analyses can be supported by the MINT data system. PMID- 11439705 TI - Earnings histories of SSI beneficiaries working in December 1997. AB - Disabling conditions previously considered to be permanent and total are no longer viewed as automatic barriers to work. Medical advances, improved accommodations in the workplace, and changes in the nature of work for the working disabled have allowed many disabled people to rejoin the workforce. The Social Security Administration (SSA) has followed those developments with a view toward encouraging people receiving disability benefits to consider returning to work. To effectively target SSA's efforts and evaluate their success, information about previous work histories of the Supplemental Security Income (SSI) beneficiary population is used to provide baseline data. This article examines the earnings histories of 300,000 disabled SSI beneficiaries--one of the populations targeted by the expanded work-incentive measure under Public Law 106 70--who were working in December 1997. The article also investigates whether beneficiaries who are working have significant lifetime earnings and whether earnings patterns exist that might assist with SSA's work-support activities. SSI program records were matched to data in the Master Earnings File to explore the characteristics and earnings patterns before and after a person applies for benefits. The article addresses several questions: What are the general characteristics of disabled SSI beneficiaries? What are their earnings histories? Did they have an earnings record when they applied for SSI? Of the SSI beneficiaries working in December 1997, most tended to be younger than other disabled beneficiaries, to have some sort of mental disability, and to have earnings well below levels that would suggest their eventual, complete independence from the SSI cash benefits program. A look at past covered earnings revealed that the vast majority of SSI workers had a history of earnings before they applied for SSI benefits. Despite their severe impairments and age at the time of first eligibility, nearly 40 percent had earnings in 11 years or more. The amounts of those earnings were quite low, however, and were usually not high enough to preclude SSI eligibility. Examining the years immediately before and after the point of application indicated whether recent pre-application earnings were consistent with post-application return to work. Results were a bit surprising. They revealed that one-third of the 1997 SSI workers had no earnings, and another 28 percent returned to work despite having no earnings in the 4-year period before application. Persons receiving SSI because of mental retardation seemed to have poorer earnings histories than other workers but were more likely to return to work after application. That may be explained by their younger ages or may reflect the outside assistance they received in responding to SSA work incentives. PMID- 11439706 TI - A benefit of one's own: older women's entitlement to Social Security retirement. AB - This article explores differences in Social Security eligibility and benefit levels for older men and women using survey data from the Health and Retirement Study combined with administrative records on actual work histories and Social Security rules. We are able to determine the fully insured status of those persons, how close they are to meeting eligibility criteria when they are not fully insured, and their prospects for benefits. Around three-quarters of older women nearing retirement today will be fully insured for Social Security old-age benefits on the basis of their own accounts, but the rest would need substantial extra employment to rise above the eligibility threshold. Further, two-thirds of older married women who are fully insured have sufficient lifetime earnings to translate into an age-65 primary insurance amount worth at least half their husband's, but the other one-third can expect no additional retirement benefit from contributing to Social Security late in life. Finally, most wives will not be able to improve their benefits by working more under current rules. These results have mixed implications regarding the potential impact of women's rising labor force attachment on eventual retirement benefits. Working more years could increase women's chances of becoming eligible for Social Security benefits, but that effect is likely to be small. Furthermore, even when women do become fully insured according to the rules, not many wives will receive a higher benefit at the margin. The reason is that married women still receive higher Social Security benefits as a spouse than they do on the basis of their own work record. In fact, the net benefit from Social Security due to additional work is negative once one takes into account the Social Security contributions the women paid while employed. Benefits paid to widows are even more likely to be based on the spouse's work history rather than on the woman's. Hence, the rising labor market attachment of women in the future may increase their eligibility for benefits but will produce only modest (and often negative) impacts on their old-age Social Security benefits under current rules. PMID- 11439707 TI - Cultural variation: considerations and implications. AB - Cultural systems vary widely across the world. Partly this is due to different cultures' occupying different ecological and environmental niches. But partly it is due to similar circumstances giving rise to multiple stable equilibriums, each with a distinct cultural form. Using insights and examples from various fields, this article illustrates the way that multiple equilibriums can emerge and the forces that push a culture toward one equilibrium point or another. Considerations of game theory principles, mutual interdependence, historical circumstance, dependence on initial conditions, and crucial choice points are highlighted in discussing the ways humans create and re-create their culture. Cultural traits develop within physical, social, intracultural, and intercultural niches, and implications of this for how culture might be studied and the benefits of combining an "equilibrium" perspective and a "meaning" perspective are discussed. PMID- 11439708 TI - Marriage and health: his and hers. AB - This review focuses on the pathway leading from the marital relationship to physical health. Evidence from 64 articles published in the past decade, particularly marital interaction studies, suggests that marital functioning is consequential for health; negative dimensions of marital functioning have indirect influences on health outcomes through depression and health habits, and direct influences on cardiovascular, endocrine, immune, neurosensory, and other physiological mechanisms. Moreover, individual difference variables such as trait hostility augment the impact of marital processes on biological systems. Emerging themes in the past decade include the importance of differentiating positive and negative dimensions of marital functioning, the explanatory power of behavioral data, and gender differences in the pathways from the marital relationship to physiological functioning. Contemporary models of gender that emphasize self processes, traits, and roles furnish alternative perspectives on the differential costs and benefits of marriage for men's and women's health. PMID- 11439709 TI - Humor, laughter, and physical health: methodological issues and research findings. AB - All published research examining effects of humor and laughter on physical health is reviewed. Potential causal mechanisms and methodological issues are discussed. Laboratory experiments have shown some effects of exposure to comedy on several components of immunity, although the findings are inconsistent and most of the studies have methodological problems. There is also some evidence of analgesic effects of exposure to comedy, although similar findings are obtained with negative emotions. Few significant correlations have been found between trait measures of humor and immunity, pain tolerance, or self-reported illness symptoms. There is also little evidence of stress-moderating effects of humor on physical health variables and no evidence of increased longevity with greater humor. More rigorous and theoretically informed research is needed before firm conclusions can be drawn about possible health benefits of humor and laughter. PMID- 11439710 TI - Status differences and in-group bias: a meta-analytic examination of the effects of status stability, status legitimacy, and group permeability. AB - This work examines the moderating effects of status stability, legitimacy, and group permeability on in-group bias among high- and low-status groups. These effects were examined separately for evaluative measures that were relevant as well as irrelevant to the salient status distinctions. The results support social identity theory and show that high-status groups are more biased. The meta analysis reveals that perceived status stability, legitimacy, and permeability moderate the effects of group status. Also, these variables interacted in their influences on the effect of group status on in-group bias, but this was only true for irrelevant evaluative dimensions. When status was unstable and perceived as illegitimate, low-status groups and high-status groups were equally biased when group boundaries were impermeable, compared with when they were permeable. Implications for social identity theory as well as for intergroup attitudes are discussed. PMID- 11439711 TI - Perceiving imitatible stimuli: consequences of isomorphism between input and output. AB - For more than a century, psychologists have been intrigued by the idea that mental representations of perceived human actions are closely connected with mental representations of performing those same actions. In this article, connections between input and output representations are considered in terms of the potential for imitation. A broad range of evidence suggests that, for imitatible stimuli, input and output representations are isomorphic to one another, allowing mutual influence between perception and motoric planning that is rapid, effortless, and possibly obligatory. Thus, the cognitive consequences of imitatibility may underlie such diverse phenomena as phoneme perception; imitation in neonates; echoic memory; stimulus-response compatibility; conduction aphasia; maintenance rehearsal; and a variety of developmental and social activities such as language acquisition, social learning, empathy, and monitoring one's own behavior. PMID- 11439712 TI - Broad and narrow perspectives in grief theory: comment on Bonanno and Kaltman (1999). AB - G. A. Bonanno and S. Kaltman's (1999) concentration on the grief work hypothesis in their review of perspectives on bereavement is only 1 aspect required for a comprehensive theory of grief, which should cover the following: the origin and adaptive significance of grief; the mechanism that initiates the grief response; the mechanism that leads to the resolution of grief; and sources of individual variation. Bonanno and Kaltman's replacement for grief work also emphasized cognitive restructuring, which can be contrasted with 2 alternative mechanisms, forgetting and engaging in new activities and relationships. M. S. Stroebe and H. A. W. Schut's (1994) dual process model, which involves cognitive restructuring and engaging in new activities, provides a broader alternative to grief work than that advocated by Bonanno and Kaltman. PMID- 11439713 TI - The crucial importance of empirical evidence in the development of bereavement theory: reply to Archer (2001). AB - In his commentary, J. Archer (2001a) argued that G. A. Bonanno and S. Kaltman's (1999) review and integration of the bereavement literature failed to consider evolutionary theory or other approaches to the origins of grief. Archer also argued that Bonanno and Kaltman had merely replaced the traditional grief work perspective with cognitive restructuring, thereby ignoring the processes related to avoidance and distancing from the loss. In this reply, the author first explains that although it was compelling to do so, Bonanno and Kaltman did not emphasize an evolutionary approach to the origins of grief reactions because in their current form these theories lack empirical and theoretical clarity. Second, the author shows that, contrary to Archer's reading, Bonanno and Kaltman's article viewed cognitive restructuring as a mechanism used primarily by extremely grieved persons and only in some cognitive domains. Last, the author shows that Bonanno and Kaltman have championed rather than ignored avoidant or distancing processes. PMID- 11439714 TI - December 2000: associated medical conditions in aviation personnel. PMID- 11439715 TI - Prevalence of hepatitis A, B, C, and E antibody in flying airline personnel. AB - OBJECTIVES: The aim of the study was to detect the prevalence of antibodies against hepatitis A, B, C, and E viruses in flying airline personnel, and to determine the necessity of hepatitis A vaccination to prevent such infections related to occupational exposure. METHODS: Antibodies against hepatitis A (HAV), B (HBV), C (HVC), and E (HEV) were tested for using standard enzyme immunoassay in airline personnel, 208 flying personnel, 199 ground crew, and 204 employees from companies not involved in travel activities. RESULTS: Prevalence of antibodies against HAV was less than 5% in each group, and there was no significant difference between the three groups. Prevalence of antibodies against HEV was significantly higher in flying personnel (3.4%) than in the control groups. Prevalence of antibodies against HBV and HCV was low in each of the three groups and there were no differences between the three groups. CONCLUSIONS: Infection with HAV, HBV and HCV does not seem to represent an occupational hazard to flying personnel. It is possible that flying personnel are exposed to infection with HEV, however, presently no vaccine is available. PMID- 11439716 TI - Airsickness and aircraft motion during short-haul flights. AB - BACKGROUND: There is little quantitative information that can be used to predict the incidence of airsickness from the motions experienced in military or civil aviation. This study examines the relationship between low-frequency aircraft motion and passenger sickness in short-haul turboprop flights within the United Kingdom. METHODS: A questionnaire survey of 923 fare-paying passengers was conducted on 38 commercial airline flights. Concurrent measurements of aircraft motion were made on all journeys, yielding approximately 30 h of aircraft motion data. RESULTS: Overall, 0.5% of passengers reported vomiting, 8.4% reported nausea (range 0% to 34.8%) and 16.2% reported illness (range 0% to 47.8%) during flight. Positive correlations were found between the percentage of passengers who experienced nausea or felt ill and the magnitude of low-frequency lateral and vertical motion, although neither motion uniquely predicted airsickness. The incidence of motion sickness also varied with passenger age, gender, food consumption and activity during air travel. No differences in sickness were found between passengers located in different seating sections of the aircraft, or as a function of moderate levels of alcohol consumption. CONCLUSIONS: The passenger responses suggest that a useful prediction of airsickness can be obtained from magnitudes of low frequency aircraft motion. However, some variations in airsickness may also be explained by individual differences between passengers and their psychological perception of flying. PMID- 11439717 TI - Results of Holter monitoring U.S. Air Force aircrew with ectopy on 12-lead electrocardiograms. AB - PURPOSE: To evaluate the results of 24-h Holter monitoring performed on healthy U.S. Air Force aircrew with asymptomatic ectopy on resting electrocardiograms (ECG). METHODS: A historical review of the USAF Central ECG Library database was conducted on all Holter studies completed for evaluation of ECG ectopy between 1 Jan 86 and 31 Dec 97. Univariate and multivariate statistical analyses were performed to determine the association between ectopy and the aeromedical dispositions of aircrew evaluated for incidental ECG ectopy. RESULTS: During this period, 147,571 resting ECGs were submitted to the Aeromedical Consultation Service (ACS) for interpretation. The mean age of the subjects was 35 yr (range 19 to 57 yr). There were 480 24-h Holter studies performed for ECG ectopy. Of these, 49% had normal or normal variant findings; another 11% were found acceptable for flying after normal treadmill testing and echocardiography. ACS evaluation was required for the remaining 40% of subjects. Overall, 4% were permanently disqualified, and 17% were lost to follow-up. Excluding subjects lost to follow-up, 95% of aircrew were returned to flying status (with or without a waiver). CONCLUSIONS: When controlled for age, no significant difference of aeromedical outcome was seen when comparing supraventricular and ventricular ectopy. Of the subjects, 51% had abnormal Holter studies, but the vast majority were returned to flying. Results of this study provide information useful in further defining aircrew medical evaluation protocols for evaluation of asymptomatic ECG ectopy with Holter monitoring. PMID- 11439718 TI - Positive predictive value of cardiac fluoroscopy in asymptomatic U.S. Army aviators. AB - The primary aim of this study was to determine the positive predictive value (PPV) of cardiac fluoroscopy in U.S. Army aviators. Cardiac fluoroscopy is one of the non-invasive tests used to screen for coronary artery disease (CAD) in the U.S. Army Cardiovascular Screening Program. The secondary objective is to determine the positive predictive value (PPV) of the combination of the aeromedical graded exercise test (GXT) and cardiac fluoroscopy. The results of these two screening tests are used to determine the need for coronary angiography. Records of 220 male aviators (mean age--42.3) who underwent coronary angiography from 1990-95 were obtained from the Aviation Epidemiology Data Register (AEDR) at Ft. Rucker, AL. These records contained results from the screening tests (GXT, cardiac fluoroscopy, and thallium scintigraphy) and coronary angiography. Significant CAD (SCAD) was present in 47 (21%), while 83 (38%) had minimal CAD (MCAD). The PPV of cardiac fluoroscopy was 81% for all CAD (34% for SCAD). GXT and thallium scintigraphy had a PPV of 62 and 67% for all CAD (23 and 45% for SCAD), respectively. The PPV's of the screening tests were not statistically different at the 95% confidence interval level. The combination of GXT and cardiac fluoroscopy had a PPV of 64% (21% for SCAD). PMID- 11439719 TI - Cardiovascular risk factors in supersonic pilots in Poland. AB - Selected cardiovascular risk factors were examined in 272 active pilots of supersonic aircraft in Poland. Hypercholesterolemia was present in 72.4% and hypertriglyceridemia in 17.1% of the pilots. Decreased levels of HDL-cholesterol and increased levels of LDL-cholesterol were found in 86.9% and 69.9% of pilots, respectively. Slightly over half (52.2%) were found to be mildly overweight while 6.6% were obese. The prevalence of smoking was 25.4%. Risk factor modification included non-pharmacological treatment of hyperlipidemia supported by lipid lowering drugs, depending on the serum lipid level. A significant improvement in lipid profiles was obtained with this strategy, at least over 3 to 6 mo of follow up. The challenge is to develop strategies that will result in maintained improvements. PMID- 11439720 TI - Prevalence of chlorpheniramine in aviation accident pilot fatalities, 1991-1996. AB - BACKGROUND: Chlorpheniramine is known to cause drowsiness, and this side effect has a potential to impair performance and could be a factor in accidents. METHODS: Therefore, to establish the prevalence of this drug in pilot fatalities of aviation accidents, a postmortem toxicology database--maintained at the Civil Aeromedical Institute--was examined for the presence of chlorpheniramine in the fatalities, occurred during 1991-1996. RESULTS: There were 47 (2.2%) accidents involving chlorpheniramine. Of these, 16 had only chlorpheniramine at 109 ng.ml-1 (n = 4) in blood and 1412 ng.g-1 (n = 12) in liver. Other drugs were also present in the remaining 31 cases, wherein chlorpheniramine concentrations were 93 ng.ml 1 (n = 18) in blood and 747 ng.g-1 (n = 12) in liver. Ninety-five percent of all quantitated blood values were at or above the therapeutic level (10 ng.ml-1), giving a 100 ng.ml-1 (n = 21) mean blood value. The drug's mean liver concentration from all cases was 1080 ng.g-1 (n = 24). The average blood value was approximately 10 times higher than the therapeutic value. CONCLUSIONS: The presence of other drugs did not appear to significantly alter the blood chlorpheniramine level, but no such correlation could be established with the hepatic value. The approximate 10-fold increase in the liver concentration was consistent with the general trend of drug distribution in the hepatic compartment. However, the contribution of postmortem redistribution of the drug to alter its concentration cannot be entirely ruled out. This study suggests that chlorpheniramine was present in some aviation fatalities at levels higher than therapeutic levels. PMID- 11439721 TI - Practical color vision tests for air traffic control applicants: en route center and terminal facilities. AB - BACKGROUND: Two practical color vision tests were developed and validated for use in screening Air Traffic Control Specialist (ATCS) applicants for work at en route center or terminal facilities. The development of the tests involved careful reproduction/simulation of color-coded materials from the most demanding, safety-critical color task performed in each type of facility. METHODS: The tests were evaluated using 106 subjects with normal color vision and 85 with color vision deficiency. The en route center test, named the Flight Progress Strips Test (FPST), required the identification of critical red/black coding in computer printing and handwriting on flight progress strips. The terminal option test, named the Aviation Lights Test (ALT), simulated red/green/white aircraft lights that must be identified in night ATC tower operations. Color-coding is a non redundant source of safety-critical information in both tasks. RESULTS: The FPST was validated by direct comparison of responses to strip reproductions with responses to the original flight progress strips and a set of strips selected independently. Validity was high; Kappa = 0.91 with original strips as the validation criterion and 0.86 with different strips. The light point stimuli of the ALT were validated physically with a spectroradiometer. The reliabilities of the FPST and ALT were estimated with Chronbach's alpha as 0.93 and 0.98, respectively. CONCLUSIONS: The high job-relevance, validity, and reliability of these tests increases the effectiveness and fairness of ATCS color vision testing. PMID- 11439722 TI - Functional magnetic resonance imaging of mental strategy in a simulated aviation performance task. AB - BACKGROUND: The objective of this study was to analyze the sensory and cognitive functions associated with activated brain regions characterizing mental strategy relative to degree of expertise in aviation-related tasks. METHODS: We used echo planar functional magnetic resonance imaging (fMRI) technique to examine brain activity in expert pilots (n = 6) compared with novice pilots (n = 6) during performance of a simulated aviation track-following task at 200 knots vs. 100 knots. RESULTS: Expert pilots showed reduced activity in visual and motor regions that contrasted with predominant activation within anterior structures including the frontal and prefrontal cortices; structures involved in visual working memory, planning, selective attention and decision making functions. Novice pilots showed widespread activation of anterior and posterior brain structures, with a rise in activity in the visual, parietal and motor cortices as task difficulty increased. CONCLUSIONS: A high level of performance in the track following task related to a high degree of expertise in the aviation field. This corresponded to experts performing perceptual and mnemonic processing through a network of specialized functions from visual through multiple prefrontal areas. By contrast, the novice pilots predominantly show activity associated with non specific perceptual processing and without subsequent representation of selective information in working memory. PMID- 11439723 TI - Power spectral analysis imperfectly informs changes in sympathetic traffic during acute simulated microgravity. AB - The purpose of this study was to investigate the value of frequency-domain analysis of autonomic rhythms as a simple, non-invasive technique for the study of immediate neural adjustments to simulated microgravity. We continuously recorded the electrocardiogram, non-invasive beat-by-beat arterial pressure, and muscle sympathetic nerve activity (MSNA) during 5-min periods of controlled frequency breathing (15 breaths.min-1) with subjects (n = 10) in supine, and 10 degrees head-down tilt positions. We estimated changes in fluid volume with lower leg circumference measurements. We analyzed data in the frequency domain with fast Fourier-based power spectral analysis, and calculated the ratio of normalized low-to-respiratory frequency RR-interval spectral power as an index of sympathetic activity. Head-down tilt significantly reduced lower leg volume, MSNA, and MSNA oscillations at the respiratory frequency (p < 0.05). Head-down tilt did not change RR-interval, arterial pressure, or their power spectra (p > 0.05). We conclude that non-invasive frequency-domain estimates do not adequately reveal subtle changes in sympathetic traffic during acute, simulated microgravity. PMID- 11439724 TI - Antiorthostatic suspension for 14 days does not diminish the oxidative response of neutrophils in mice. AB - The effects of long-term spaceflight on inflammatory responses have not been well studied in either humans or animals. It is thus important to determine if the functions of immune and inflammatory cells are altered in models of spaceflight. One such animal model is antiorthostatic suspension (AOS), in which the experimental animal is subjected to a head-down tilt that mimics both the stress and the cephalad fluid shift experienced in spaceflight. A previous study reported that the peritoneal neutrophils from mice experiencing AOS generated less superoxide than unsuspended controls. We expanded on this study using several different stimuli and measuring the oxidative response of murine neutrophils in a variety of ways. These responses included the rate, lag period, and dose/response characteristics for superoxide generation, FACS analysis with dihydrodichlorofluorescein as a substrate, and a chemiluminescence response with luminol as a substrate. We also examined phagocytosis of three different microorganisms. While some effects of orthostatic suspension (attributable to the stress of the apparatus) were observed, no clear effects of AOS on oxidative function of the peritoneal neutrophils were seen. PMID- 11439725 TI - Exposure to soda-lime dust in closed and semi-closed diving apparatus. AB - BACKGROUND: Chronic exposure to hyperbaric hyperoxia and venous gas microembolism have been shown to contribute to the long term health effects of diving, especially diver's lung function. Factors related to special diving equipment may add to these effects. This study was conducted to evaluate possible additional hazards for respiratory function of divers employing closed and semi-closed diving apparatus. METHODS: We analyzed soda-lime dust found in the air-intake loop of a closed-circuit oxygen rebreathing diving apparatus which had passed through the filter screen of the diving apparatus's soda-lime cartridge. The geometrical characteristics were evaluated by scanning electron microscopy. The amount of dust that passed through the screen during a normal dive profile was measured by an artificial airflow through a filter membrane. After dives by subjects using these devices, the pH-value of water condensate in the air-intake hose was measured. RESULTS: There was a relevant amount of residual soda-lime dust found in the air-intake loop. The dust particles showed diameters down to 1 micron and a rounded structure. The total amount of dust averaged 9.6 mg.m-3 of breathing mixture. During diving, the mean pH-value of condensate in the hose is estimated at 8.87 (+/- 0.12). CONCLUSION: There is a relevant exposure to soda lime dust in divers using closed-circuit rebreathing apparatus. This occupational exposure may contribute to chronic airway inflammation and subsequent development of small airway disease in divers. PMID- 11439726 TI - An airman with tuberculous uveitis: case study. AB - Tuberculosis is endemic, with nearly 2 billion carriers worldwide. Aviation medical examiners should be alert to both its pulmonary and extrapulmonary manifestations, especially in the international traveler. Uveitis is a rare presentation of tuberculosis, but that diagnosis must be considered in the differential, even in the absence of pulmonary disease. Promptly treated, tuberculosis should not interfere with the resumption of a career in aviation, and does not generally require special follow up. PMID- 11439727 TI - You're the flight surgeon. Considering the differential diagnosis of seizures, a flight surgeon does not need an exhaustive list but rather a focus on the most common causes. PMID- 11439728 TI - You're the flight surgeon. Possible causes of fatigue and a slight right-sided abdominal discomfort. PMID- 11439729 TI - Roll vs. pitch rotation. PMID- 11439730 TI - Health plans' use of asthma quality improvement projects to meet NCQA accreditation standards. AB - BACKGROUND: Managed care represents an important system of healthcare delivery in the United States and the primary source of care for many persons with asthma. OBJECTIVE: To characterize how US managed care health plans address the quality of asthma care through the use of disease-specific quality improvement (QI) programs complying with National Committee for Quality Assurance (NCQA) standards. METHODS: This study was a cross-sectional review of reports from all accreditation surveys conducted in 1996 and 1997 by the NCQA. Each accreditation report was reviewed for evidence of whether the health plan explored asthma care as a way to demonstrate compliance with NCQA accreditation standards. Asthma activity was examined with respect to health plan characteristics such as size of plan and Medicaid contracting. Types of asthma QI activity, use of guidelines, and application of different NCQA accreditation standards were also examined. RESULTS: Approximately 90% of 197 health plans undergoing NCQA accreditation surveys in 1996 and 1997 reported some form of asthma QI activity. There were no statistically significant differences in this activity in large vs small plans or in plans with vs without Medicaid members. Approximately two thirds of health plans used asthma QI activities to meet NCQA accreditation standards in health management systems, and three fifths monitored and evaluated important aspects of asthma care and service. CONCLUSIONS: During the study period, many US health plans conducted asthma care QI activities. The recently released NCQA asthma performance measure may provide the next assessment of how well managed care is contributing to improving asthma care in the United States. PMID- 11439731 TI - Does the HEDIS asthma measure go far enough? PMID- 11439732 TI - Diagnosis and care of irritable bowel syndrome in a community-based population. AB - OBJECTIVE: To identify the healthcare utilization and evaluation in a community based population with an incident diagnosis of irritable bowel syndrome (IBS). STUDY DESIGN: Retrospective cohort. PATIENTS: A randomly selected cohort of 149 Olmsted County, MN, adults with an incident diagnosis of IBS between June 1, 1992, and December 31, 1994. METHODS: Retrospective medical record review of each medical encounter for the 10 years before and the 3 years after the incident IBS diagnosis. Specific attention was given to healthcare visits, tests, and treatment associated with documented gastrointestinal (GI) tract symptoms and diagnosis of IBS. RESULTS: Of 149 patients, 98 (66%) were women, and the mean patient age was 46.6 years. Patients averaged 4.7 healthcare visits annually, including 0.5 related to GI tract symptoms. Two thirds of patients had GI tract symptom-related visits at least 2 years before IBS diagnosis. Only 5% of IBS diagnoses were made by a gastroenterologist. Colon examinations (colonoscopy or barium enema) were performed on 47% of patients and were more common in those aged > or = 55 years (58% vs 36%; P = .02). One third of patients had no GI tract testing proximal to the IBS diagnosis. Following the IBS diagnosis, only about half the patients made any IBS or GI tract symptom-related visits per year. CONCLUSION: The path to diagnosis of IBS is through the primary care physician and includes minimal testing and limited follow-up care. PMID- 11439733 TI - Prevalence and determinants of osteoporosis drug prescription among patients with high exposure to glucocorticoid drugs. AB - OBJECTIVE: To investigate use of osteoporosis drugs among patients with high exposure to glucocorticoid drugs. STUDY DESIGN: Retrospective review of pharmacy records. METHODS: We identified patients aged > or = 20 years who received prescriptions for > or = 2 g of prednisone (or equivalent) during any 12-month period between January 1, 1998, and December 31, 1999, and who initiated use of osteoporosis-specific drugs (alendronate sodium, etidronate disodium, and calcitonin) during that period. RESULTS: Among 8807 patients who met study criteria, 772 (8.8%) received prescriptions for osteoporosis drugs. Prevalence of osteoporosis drug prescriptions increased linearly during the study and differed markedly by patient sex, age, and exposure to glucocorticoid drugs. Osteoporosis drugs were prescribed for 16.3% of women aged > or = 65 years, for 6.1% of women aged < 50 years, for 6.5% of men aged > or = 65 years, and for 2.2% of men aged < 50 years. Higher glucocorticoid exposure was also associated with higher rate of osteoporosis drug prescription (11.2% of patients exposed to > 4 g/y and 5.6% exposed to 2 to 3 g/y received such therapies). Osteoporosis drugs were 50% more likely to be prescribed by clinicians who prescribed glucocorticoid drugs to > 18 patients than by providers who prescribed glucocorticoid drugs to < 4 patients. CONCLUSIONS: Despite ready availability of bone-specific osteoporosis drugs, few patients with high exposure to glucocorticoid drugs received such therapy. Likelihood of an osteoporosis drug being prescribed for such patients strongly depends on patient sex, age, and exposure to glucocorticoid drugs and on level of practitioner experience in prescribing glucocorticoid drugs. PMID- 11439734 TI - Effect of rofecoxib therapy on measures of health-related quality of life in patients with osteoarthritis. AB - BACKGROUND: Bodily pain and physical disability can negatively impact health related quality of life (HRQL) in patients with osteoarthritis (OA). OBJECTIVE: To assess the effects of treatment with a new agent, rofecoxib, on HRQL in patients with OA. STUDY DESIGN: Randomized, double-blind, 6-week clinical trial comparing treatment with rofecoxib, 5 to 50 mg, with placebo in 672 patients with OA of the hip or knee. MAIN OUTCOME MEASURE: Patient HRQL was assessed at baseline and at the end of treatment using the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36). RESULTS: At 6 weeks, mean change from baseline in all SF-36 mental and physical health domain scores demonstrated significant improvement with rofecoxib use (P < .05 for all doses for all SF-36 domains), with evidence of a dose-response relation. Improvements in mental and physical HRQL domains with rofecoxib treatment were significantly greater than those with placebo treatment (P < .05 for each dose of rofecoxib vs placebo for all domains except general health) and highly correlated with improvements observed using disease-specific OA outcome measures such as the Western Ontario and McMaster Universities Osteoarthritis Index-visual Analog 3.0 OA index pain and physical function subscales. The effect of rofecoxib vs placebo treatment on mental health largely disappeared after adjustment for improvement in OA disease-specific measures. CONCLUSIONS: Rofecoxib treatment increased physical and mental HRQL domain scores on the SF-36. Improvements in mental health with rofecoxib use primarily resulted from effective treatment of OA (i.e., reduction in pain and improvement in physical function). PMID- 11439735 TI - Rheumatoid arthritis: guidelines for emerging therapies. AB - The individual and societal impacts of rheumatoid arthritis are of substantial consequence. Management of the disease has pharmacologically focused on the use of anti-inflammatories and disease-modifying antirheumatic drugs, which are only partially successful in retarding joint destruction and functional disability. The recent emergence of cytokine antagonists (anti-tumor necrosis factor therapy) challenges clinicians and managed care organizations with the need to develop new treatment guidelines. Recent developments in the understanding of rheumatoid arthritis, including its epidemiological characteristics, economic costs, clinical progression, and current and emerging therapies, are reviewed. Pharmacologic utilization models are proposed. Pending the development of broad based consensus treatment recommendations, interim treatment guidelines are suggested. PMID- 11439736 TI - Resolution of renal disease: mission impossible? AB - Several studies have extensively shown that both dietary and pharmacological intervention can prevent the progression of renal damage. The best results may be obtained by optimizing blood pressure control, reducing proteinuria levels in non diabetic nephropathies, and further achieving a good glycemic control in diabetic nephropathies. The earlier the treatment is started, the better the results. Since slowing progression of renal disease has been established, the challenge of the future seems to be the resolution of an established renal damage. Few studies have suggested that this process of regression is possible. Experimental animal studies, based on repeated renal morphological investigations, showed resolution of glomerular lesions in 40% of animals treated with either ACEI or AIIRA. Resolution of renal lesions (62%) has been claimed in a single study and in a small number of patients with diabetic nephropathy after 10-year pancreas transplantation. Confirmation studies are awaited. PMID- 11439737 TI - Vascular access for hemodialysis: an indepth review. AB - A multi-disciplinary approach to vascular access planning, early preoperative surgical assessment and increased use of vascular imaging for surveillance, specialist service and pharmacological prevention of intimal hyperplasia will lead to improved patency rates for both autologous AVF and synthetic grafts. PMID- 11439738 TI - Effect of sodium pool changes on blood pressure in patients undergoing PFD: design of a prospective randomized multicenter trial. AB - Blood pressure control is important during dialysis and the interdialytic period because of the frequency and potential seriousness of hypotension and hypertension. Water and sodium removal play an important role in the genesis of intradialytic cardiovascular instability or hypertension. Changing dialysate sodium concentrations without the aid of a kinetic model can sometimes give good results but is only an empirical approach. Therefore, this clinical trial was designed to prospectively investigate the advantages of changes in the sodium pool on the blood pressure profile of patients undergoing paired filtration dialysis (PFD). The hypothesis to be tested is whether using a dialysate conductivity which, according to the conductivity kinetic model, ensures that the conductivity of the ultrafiltrate at the end of each dialysis session is 0.3 mS/cm more (B) or less (C) than the mean during the run-in period, improves blood pressure control either in patients prone to intradialytic hypotension or patients who are hypertensive or normotensive with antihypertensive treatment. Patients will be randomly allocated to one of two treatment sequences (where treatment A is standard PFD): AABB or ABAA for patients with intradialytic hypotension; AACC or ACAA for hypertensive patients. During the experimental phase arterial blood pressure will be measured and symptoms reported by the patients will be recorded. PMID- 11439739 TI - Modelling the "ideal" self care--limited care dialysis center. AB - Limited care dialysis is an interesting option, which has gained attention in several settings because of the aging of the uremic cohort. The aim of this study was to assess its potential in the Piedmont region in northern Italy, evaluating patients' and care-givers' preferences and testing them in a mathematical model of organisation. The study was conducted in the satellite unit of a university hospital (200-210 dialysis patients), following 35 patients (15 at home, 20 in the center, 10 on daily dialysis). Opinions were collected with a questionnaire and features identified were empirically tested through a simulation model. Most patients (34/35) preferred a small unit, with a stable caring team. Further options were flexibility of dialysis schedule, multiple treatment options, integrated center/home care. These needs could be met by a flexible organization including conventional dialysis (3/week) and daily dialysis (6/week). We employed a simulation model (ARENA software) to calculate the nurses required for each shift and the opening hours and best schedule for the unit. Addition of daily dialysis (2-3 hours) to two conventional 4-5 hour sessions to increased the number of patients followed or "spared" beds, ensuring flexibility. According to patients' best choice (7 dialysis stations), and to the recorded calls, the needs are for two nurses per shift, two shifts per day and six nurses for up to 30 patients in limited care. In conclusion, small centers with flexible schedules can tailor dialysis to patients' needs. A managerial approach is valuable for testing cost/benefit ratios in specific contexts. PMID- 11439740 TI - Rubidium deficiency in dialysis patients. AB - BACKGROUND: Since dialysis has brought long-term survival to uremic patients, we can now speculate on more subtle problems derived from imbalance or sub-optimal regulation of some elements such as trace metals. We focused on the rubidium (Rb) status in dialysis patients (HD), as concerns about its possible deficiency have been raised. METHODS: Rb in uremic patients was evaluated by: A) serum concentration (graphite furnace atomic absorption spectroscopy) from blood samples of 70 patients on chronic hemodialysis (HD) in comparison with 75 controls; B) tissue concentration (neutron activation analysis) from autopsy or biopsy samples (20) of HD patients in comparison with 21 controls; C) in vivo intradialytic mass balance during standard bicarbonate dialysis in 8 HD patients. RESULTS: A) Serum Rb concentrations in HD patients significantly were lower than in normal controls (304 +/- 81 micrograms/L versus 350 +/- 74 micrograms/L p < 0.001, log-transformed 5.68 +/- 0.28 versus 5.84 +/- 0.20, p < 0.001). Univariate logistic regression analysis found a significantly higher risk of serum Rb < 250 300 and 350 micrograms/L in uremic patients than in controls (Odd ratios or 12.6, 95% CI 2.77-57.04; 4.0, 95% CI 1.92-8.4; 2.08, 95% CI 1.02-4.25, respectively). B) Rb was significantly lower in tissues of HD patients, including brain (2250 +/ 1520 ng/g versus 5490 +/- 1250 ng/g, p = 0.0002) than normal controls. C) Rb was transferred from the patients' blood to the dialysis bath during a standard bicarbonate dialysis session, giving mean intradialytic Rb removal of 4.0 +/- 1.1 mg/session. CONCLUSIONS: These results confirm that Rb deficiency may arise in uremic patients, and indicate that diffusive dialysis treatments allow Rb removal which, however, with a standard bicarbonate schedule does not seem to be any greater than that expected with normal urine output (20 mg/week). Further studies are needed to clarify the roles of many factors in this Rb deficiency, including the effects of uremia by itself, pre-dialysis factors (diet, impaired renal function and drugs), dialysis procedures (frequency, hours, diffusive/convective components) or other biochemical/clinical parameters (hemoglobin, body mass index, age). The finding of a Rb deficiency in uremia is important as it has a role in neurobehavioural functions, mainly as an antidepressant. As Rb deficiency may be implicated in central nervous system alterations which strongly influence the quality of life, we believe that monitoring serum Rb in uremic patients and clarifying the causal mechanisms of deficiency will facilitate future therapeutic approaches. PMID- 11439741 TI - Sevelamer reduces calcium load and maintains a low calcium-phosphorus ion product in dialysis patients. AB - BACKGROUND: Sevelamer HCl, a non-aluminum, non-calcium containing hydrogel, has proved an effective phosphate binder in North American hemodialysis patients. This single-center, open-label, dose titration study assessed the efficacy of sevelamer in a cohort of European hemodialysis patients with different dietary habits, in particular with lower phosphate intake. The aim of the study was to obtain a calcium x phosphate product lower than 60 mg2/dL2 in all patients. METHODS: Administration of calcium- or aluminum-based phosphate binders was discontinued during a two-week washout period. Nineteen patients whose serum phosphate level at the end of washout was greater than 5.5 mg/dL (1.78 mmol/L) qualified to receive sevelamer for six weeks. Based on the degree of hyperphosphatemia during washout, patients were started on 403 mg sevelamer capsules with a dose schedule different from previous studies. Only one capsule was administered at breakfast, and the rest of the phosphate binder was divided equally at the two main meals. Sevelamer could be increased by two capsules per day every two weeks, if necessary. A second two-week washout period followed. RESULTS: Mean serum phosphorus rose from a baseline of 5.3 +/- 1.0 to 7.4 +/- 1.4 mg/dL at the end of washout, then declined to 5.4 +/- 0.8 mg/dL (p < 0.001) by the end of the six-week treatment period and rebounded significantly to 7.1 +/- 1.1 mg/dL after the second two-week washout. Calcium x phosphate product showed a similar pattern, decreasing significantly from 64.1 +/- 14.1 to 46.9 +/- 7.4 mg2/dL2 (p < 0.001) after six weeks of sevelamer. A level of less than 50 mg2/dL2 was reached by 68% of patients, and 95% had less than 60 mg2/dL2. The mean dose of sevelamer at the end of treatment was 3.1 +/- 0.6 g per day. As expected, calcium declined from 9.2 +/- 0.5 to 8.7 mg/dL (p < 0.01) during the initial washout after stopping calcium-based phosphate binders, but remained stable thereafter. Ionized calcium did not change significantly throughout the washout and sevelamer treatment. However, interruption of calcium salts led to a 81% reduction of total calcium intake. CONCLUSIONS: We confirmed in an European sample of hemodialysis patients that sevelamer can reduce phosphate levels without inducing hypercalcemia. The drug can also be successfully used to reduce mean calcium x phosphate levels below 50 mg2/dL2, closer to normal values. Although similar results can be obtained with other phosphate binders, a concomitant accumulation of aluminum, calcium or magnesium could be detrimental to patients. PMID- 11439742 TI - By reducing TGF beta 1, octreotide lessens the peritoneal derangements induced by a high glucose solution. AB - BACKGROUND: Chronic peritoneal dialysis may eventually result in peritoneal fibrosis, which progressively reduces dialytic efficacy. Although the pathogenesis has not been elucidated, it has been proposed that transforming growth factor beta-1 (TGF beta 1) plays a central role in the onset of peritoneal fibrosis. METHODS: Rats were divided into three groups and given saline, hypertonic peritoneal dialysis solution alone, a hypertonic peritoneal dialysis solution plus octreotide intraperitoneally. After four weeks, a one-hour peritoneal equilibration test was done. Dialysate-to-plasma urea ratio, glucose reabsorption, ultrafiltration volume and levels of dialysate protein, TGF beta 1 and cancer antigen 125 (CA 125) were determined. The peritoneal membrane was examined histologically by light microscopy. RESULTS: Compared to the saline group, peritoneal function tests (ultrafiltration volume 6 (5-7) vs 0.0 ml, dialysate-to-plasma urea ratio 0.51 vs 0.76, glucose reabsorption 0.54 vs 0.40 and morphology (thickness 4.5 vs 75.5 microns) were dramatically deranged in hypertonic peritoneal dialysis solution-treated rats, which also had a higher level of TGF beta 1 and undetectable CA 125. In contrast, in hypertonic peritoneal dialysis solution plus octreotide rats' peritoneal function was protected (ultrafiltration volume 3 mL, dialysate-to-plasma urea 0.60, glucose reabsorption 0.51) but peritoneal thickening (37.7 microns) was not so markedly reduced although the production of TGF beta 1 was significantly inhibited. CONCLUSION: These data show that by inhibiting the production of TGF beta 1, octreotide can preserve peritoneal function and remodeling of the mesothelial cell. Although the production of TGF beta 1 was significantly inhibited, peritoneal thickening cannot be completely prevented. PMID- 11439743 TI - Cortical fibroblast culture from human biopsies. AB - BACKGROUND: Tubulointerstitial fibrosis is an integral part of progressive renal disease. Human cortical fibroblasts are believed to be key effector cells in fibrogenesis. Thus, a reliable culture of these cells is necessary for studies of their pathophysiology. METHODS: Cortical fibroblast culture from routine kidney biopsies were analyzed and the cells were characterized. Indirect immunofluorescence staining was done after the first passage for cytokeratin, vimentin, alpha-smooth muscle actin, CD 44, CD 54, CD 68, collagen types I, III, and HLA-DR. We then assessed the utility of the putative fibroblast markers CD 90, prolyl-4-hydroxylase (P4H) and F1b in simultaneous stainings of tubular epithelial cells. RESULTS: During the study period, 49 biopsy cores were cultured and cortical fibroblasts could be successfully established in 21 cases (42.9%). There was no relation between the success rate of culture and the degree of interstitial fibrosis, but an association was seen with the time of completion of the first passage. There was a negative correlation between the extent of scarring and the percentage of cytokeratin positive cells (r = -0.66, p < 0.001). All primary fibroblasts were negative for factor VIII, HLA-DR, CD 68, and cytokeratin. They expressed alpha-smooth muscle actin and collagen types I and III to variable degrees. There was a robust correlation between the percentage of alpha-smooth muscle actin positive cells and interstitial scarring but no such association with collagen type I or type III positive cells. The three putative fibroblast markers did not prove useful in differentiating between tubular epithelial cells and fibroblasts. However, since only fibroblasts stained positive for CD 90 and negative for cytokeratin, these two markers may suffice to distinguish fibroblasts from other renal cellular elements. CONCLUSIONS: Cortical renal fibroblasts can be easily cultured from kidney biopsy cores, though the success rate of pure cultures is below 50%. Staining for CD 90 and cytokeratin may suffice for initial characterization of these cells. PMID- 11439744 TI - Mesangial cell cultures. AB - Cell culture of glomerular mesangial cells (MC) has been available to most renal research laboratories since the early 80s. Key to a large number of studies on the biochemistry and molecular biology of the glomerulus, MC in culture have extensive analogies with this in vivo rather undifferentiated intercapillary cell population. They proliferate in response to mitogens and growth factors, can be growth-arrested by withdrawal of serum or 3D culture in collagen gels, synthesize an extracellular matrix that includes interstitial collagens, and display most markers of mesangial origin, including a functional contractile apparatus. As proliferation and matrix synthesis/degradation in vitro are regulated by cytokines and growth factors, cultured cells are an ideal tool for studying pathophysiological events such as mesangial expansion, scarring, and glomerulosclerosis. Current techniques for MC isolation and culture are reviewed, with several methodological issues relevant to the characterization, propagation and long-term maintenance of functional clones. PMID- 11439745 TI - Isolation, culture, characterization and use of human renal tubular epithelial cells. AB - Tubulointerstitial alterations are a hallmark of most renal diseases. In vitro culture of different cell types of the tubulointerstitial compartment have been instrumental in studying their respective contributions to the inflammatory and fibrotic process. The present paper provides an overview of the isolation, culture and characterization of tubular epithelial cells, with a main emphasis on cells from human origin. PMID- 11439746 TI - Altering glomerular epithelial function in vitro using transient and stable transfection. AB - Damage of the glomerular filtration barrier leads to proteinuria and progressive renal failure. Several independent lines of research have implicated the glomerular epithelial cell (GEC) as a key player in initiation and propagation of pathways leading to glomerulosclerosis. A growing number of molecules activated in this process have been identified. To further define their cellular function, manipulation of these molecules using pharmacological or genetic approaches in tissue culture systems are required. In this study, strategies for altering GEC gene expression by transient and stable transfection of fluorescence labeled proteins will be presented and discussed. The insight gained through these and comparable systems should allow a detailed dissection of the molecular pathways active in GEC function and failure. PMID- 11439747 TI - Articles related to high-functioning individuals with autism or Asperger disorder. PMID- 11439748 TI - Brief report: case reports on naltrexone use in children with autism: controlled observations regarding benefits and practical issues of medication management. PMID- 11439749 TI - Brief report: effects of clozapine on self-injurious behavior of two risperidone nonresponders with mental retardation. AB - Atypical antipsychotic medications for self-injurious behavior (SIB), aggression, and destruction among people with mental retardation and development disabilities are becoming increasingly accepted. Most studies are on risperidone and fewer have been conducted on clozapine. The present single-blind study reports marked reductions in SIB and aggression of two persons with profound mental retardation who were nonresponsive to all other behavioral and psychopharmacological interventions, including risperidone. The most effective dose was 200 mg/day. Side effects were mild and the drug was tolerated well. PMID- 11439750 TI - Autistic disorder and schizophrenia: diagnostic overlaps. AB - Data on 14 males with autism and 14 with schizophrenia were collected to examine symptom overlap. The Structured Clinical Interview (SCID), the schedule for positive symptoms (SAPS) and the schedule for negative symptoms (SANS) of schizophrenia, the Childhood Autism Rating Scale (CARS), and the DSM-III-R were administered. On the SCID, none of the men with paranoid schizophrenia met criteria for autism while 7 of those with autism met criteria for schizophrenia, disorganized type, showing negative symptoms. In addition, 5 showed positive symptoms on the SAPS and 6 negative symptoms on the SANS. As the difference in measured nonverbal intelligence was not significant, the effects could not be attributed to it. Although the findings continue to support the differentiation of autism and schizophrenia, they are also consistent with a comorbidity of the two disorders, mainly in those diagnosed with autism. PMID- 11439751 TI - Advanced theory of mind in high-functioning adults with autism. AB - Twenty-four high-functioning adults with autism (16 men) who passed a first-order theory-of-mind task and 24 nonautistic adults (10 men) attributed mental states to recordings of various verbal intonations and to photos of people's eyes to assess advanced theory of mind. Participants with autism performed significantly worse than nonautistic participants on both tasks. Thus, the previously described inattention to others' eyes exhibited by adults with autism is not solely responsible for their inability to attribute mental states from eyes, as they also did not correctly attribute mental states from voices. These findings support the view that a core deficit for people with autism lies in their theory of mind, that is, their inability to attribute mental states to others. PMID- 11439752 TI - Motion and emotion: a novel approach to the study of face processing by young autistic children. AB - The specificity of facial processing impairment in autistic children, particularly in the domain of emotion, is still debated. The aim of our study was to assess the influence of motion on facial expression recognition in young autistic children. Thirteen autistic children (M age: 69.38 months) were matched for gender and developmental level with a control group of 13 normal children (M age: 40.53 months). They were compared on their ability to match videotaped "still," "dynamic," and "strobe" emotional and nonemotional facial expressions with photographs. Results indicate that children with autism do not perform significantly worse than their controls in any of our experimental conditions. Compared to previous studies showing lower performance in autistic than in control children when presented with static faces, our data suggest that slow dynamic presentations facilitate facial expression recognition by autistic children. This result could be of interest to parents and specialists involved in education and reeducation of these children. PMID- 11439753 TI - Inferential language in high-function children with autism. AB - Despite average verbal intelligence, high-function children with autism have social comprehension deficits that are expressed by how they use and understand language. In this paper, we explored the general hypothesis that high-function children with autism make some, but not all, of the pragmatic inferences necessary for successful communication, even when they have the ability to perform noninferential language tasks. We contrasted the ability of 8 high function children with autism (each with Verbal IQ > 70) and typically developing children to use and understand: pragmatic inferences about given or presupposed knowledge in mental state words; pragmatic inferences about new or implied knowledge in mental state words; bridging inferences essential for coherence; elaborative inferences involved in enriching a communication by means of figurative language; and the intentional inferences involved in speech acts. High function children with autism could define words and identify multiple meanings for ambiguous words. In understanding words for mental states, they made inferences from mental state verbs to given or presupposed knowledge. However, they failed to infer what mental state verbs implied in context; to make inferences about social scripts; to understand metaphor; and to produce speech acts, all of which are inferences that are the basis of successful social communication because they elaborate meaning or convey intentions. PMID- 11439754 TI - The autism-spectrum quotient (AQ): evidence from Asperger syndrome/high functioning autism, males and females, scientists and mathematicians. AB - Currently there are no brief, self-administered instruments for measuring the degree to which an adult with normal intelligence has the traits associated with the autistic spectrum. In this paper, we report on a new instrument to assess this: the Autism-Spectrum Quotient (AQ). Individuals score in the range 0-50. Four groups of subjects were assessed: Group 1: 58 adults with Asperger syndrome (AS) or high-functioning autism (HFA); Group 2: 174 randomly selected controls. Group 3: 840 students in Cambridge University; and Group 4: 16 winners of the UK Mathematics Olympiad. The adults with AS/HFA had a mean AQ score of 35.8 (SD = 6.5), significantly higher than Group 2 controls (M = 16.4, SD = 6.3). 80% of the adults with AS/HFA scored 32+, versus 2% of controls. Among the controls, men scored slightly but significantly higher than women. No women scored extremely highly (AQ score 34+) whereas 4% of men did so. Twice as many men (40%) as women (21%) scored at intermediate levels (AQ score 20+). Among the AS/HFA group, male and female scores did not differ significantly. The students in Cambridge University did not differ from the randomly selected control group, but scientists (including mathematicians) scored significantly higher than both humanities and social sciences students, confirming an earlier study that autistic conditions are associated with scientific skills. Within the sciences, mathematicians scored highest. This was replicated in Group 4, the Mathematics Olympiad winners scoring significantly higher than the male Cambridge humanities students. 6% of the student sample scored 32+ on the AQ. On interview, 11 out of 11 of these met three or more DSM-IV criteria for AS/HFA, and all were studying sciences/mathematics, and 7 of the 11 met threshold on these criteria. Test retest and interrater reliability of the AQ was good. The AQ is thus a valuable instrument for rapidly quantifying where any given individual is situated on the continuum from autism to normality. Its potential for screening for autism spectrum conditions in adults of normal intelligence remains to be fully explored. PMID- 11439755 TI - Cognitive styles in high-functioning adolescents with autistic disorder. AB - This study addressed the operationalization, the identification, and the prevalence of weak central coherence and poor cognitive shifting in 35 high functioning adolescents with autism. Central coherence and cognitive shifting were represented by two factors in a factor analysis, each reflecting a constituent aspect of the domain in question. With regard to central coherence, these aspects were the ability of piecemeal processing and the ability to process meaning. The aspects related to cognitive shifting concerned internally and externally controlled shifting. Weak central coherence and poor cognitive shifting did not appear to be related to measures of symptom severity, social understanding, and social competence. Both these cognitive styles did not appear to be universal to autism. In our sample, weak central coherence and poor cognitive shifting were found to be significantly more common than in normative control subjects. PMID- 11439756 TI - Taking a closer look at functional play in children with autism. AB - Research evidence indicates that children with autism may experience problems with functional play, in addition to their well-documented deficits in symbolic play. However, as a result of the tendency of previous studies to group all functional play into a single category, the precise nature and extent of this deficit remains unclear. The present study undertook a more refined analysis of such play, subtyping the functional acts into various categories, in terms of the developmental progression suggested by research with typical infants. The functional play of children with autism was compared to that of developmentally matched children with Down syndrome and typical infants. Although there were no group differences in overall measures of the proportion of total play time spent in functional play and in the number of functional acts performed, a closer analysis of the composition of this play did reveal striking, qualitative differences. The functional play of the autism group was less elaborated, less varied, and less integrated than that of the controls. The implications of these findings are explored in relation to current theoretical models of autism and in relation to the role of other people in mediating the appropriate use of objects. PMID- 11439757 TI - Movement preparation in high-functioning autism and Asperger disorder: a serial choice reaction time task involving motor reprogramming. AB - Autism and Asperger disorder have long been associated with movement abnormalities, although the neurobehavioural details of these abnormalities remain poorly defined. Clumsiness has traditionally been associated with Asperger disorder but not autism, although this is controversial. Others have suggested that both groups demonstrate a similar global motor delay. In this study we aimed to determine whether movement preparation or movement execution was atypical in these disorders and to describe any differences between autism and Asperger disorder. A simple motor reprogramming task was employed. The results indicated that individuals with autism and Asperger disorder have atypical movement preparation with an intact ability to execute movement. An atypical deficit in motor preparation was found in Asperger disorder, whereas movement preparation was characterized by a "lack of anticipation" in autism. The differences in movement preparation profiles in these disorders were suggested to reflect differential involvement of the fronto-striatal region, in particular the supplementary motor area and anterior cingulate. PMID- 11439758 TI - A family genetic study of autism associated with profound mental retardation. AB - We sought to determine if the family loading for either the broader autism phenotype or for cognitive impairment differed according to whether or not autism was accompanied by severe mental retardation. The sample comprised 47 probands with autism meeting ICD-10 criteria, as assessed by the Autism Diagnostic Interview and the Autism Diagnostic Observation Schedule. Family history interview and findings were compared with those for the higher IQ autism and Down syndrome samples in the Bolton et al. (1994) study. The familial loading for autism and for the broader phenotype was closely comparable to that in the study of higher IQ autism, and different from that for Down syndrome. The family loading for scholastic achievement difficulties was slightly, but significantly, higher when autism was accompanied by severe retardation. PMID- 11439759 TI - Brief report: macrographia in high-functioning adults with autism spectrum disorder. AB - The initial description of Asperger syndrome commented on the poor handwriting and motor coordination difficulties of individuals with this condition. Early descriptions of autism do not remark upon such difficulties. Recent evidence, however, suggests that individuals with both conditions have a similar motor control impairment. Handwriting has not been formally assessed in this context. Our study compared handwriting size between individuals with autism spectrum disorder and age- and IQ-matched control subjects. Macrographia was observed among subjects with autism spectrum disorder which remained statistically significant when covaried with educational level. This finding may correlate with the anatomical abnormalities present in the cerebellum of individuals with autism spectrum disorder. PMID- 11439760 TI - Studies assess economic, environmental effects of antimicrobial drug use on hog farms. PMID- 11439761 TI - Caterpillars, cherry trees may take blame for foal deaths in Kentucky. PMID- 11439762 TI - Foot-and-mouth on the decline in United Kingdom. PMID- 11439763 TI - What is your diagnosis? Soft-tissue opacity in the right perineal region. PMID- 11439764 TI - Anesthesia case of the month. Inadvertent endobronchial intubation resulted in hypoxemia. PMID- 11439765 TI - Determining the value of a veterinary practice. PMID- 11439766 TI - Communications, privilege, and the veterinarian. PMID- 11439767 TI - Bacterial microflora of normal and telangiectatic livers in cattle. AB - OBJECTIVE: To identify potential bacterial pathogens in normal and telangiectatic livers of mature cattle at slaughter and to identify consumer risk associated with hepatic telangiectasia. SAMPLE POPULATION: 50 normal livers and 50 severely telangiectatic livers. PROCEDURE: Normal and telangiectatic livers were collected at slaughter for aerobic and anaerobic bacterial culture. Isolates were identified, and patterns of isolation were analyzed. Histologic examination of all livers was performed. RESULTS: Human pathogens isolated from normal and telangiectatic livers included Escherichia coli O157:H7 and group-D streptococci. Most livers in both groups contained bacteria in low numbers; however, more normal livers yielded negative culture results. More group-D streptococci were isolated from the right lobes of telangiectatic livers than from the left lobes, and more gram-negative anaerobic bacteria were isolated from left lobes of telangiectatic livers than from right lobes. All telangiectatic lesions were free of fibrosis, active necrotizing processes, and inflammation. CONCLUSIONS AND CLINICAL RELEVANCE: The USDA regulation condemning telangiectatic livers is justified insofar as these livers contain more bacteria than normal livers do; however, normal livers contain similar species of microflora. Development of telangiectasia could not be linked to an infectious process. The finding of E coli O157:H7 in bovine livers suggests that information regarding bacterial content of other offal and muscle may identify sources of this and other potential foodborne pathogens and assist in establishing critical control points for the meat industry. PMID- 11439768 TI - Applications of local antimicrobial delivery systems in veterinary medicine. PMID- 11439769 TI - Effect of gonadectomy on subsequent development of age-related cognitive impairment in dogs. AB - OBJECTIVE: To determine whether gonadectomy predisposes dogs to development of age-related behavioral changes linked to cognitive impairment. DESIGN: Cohort study. ANIMALS: 29 sexually intact male dogs, 63 spayed female dogs, and 47 castrated male dogs 11 to 14 years old. PROCEDURE: Information on possible impairments in 4 behavioral categories linked to cognitive impairment (orientation in the home and yard, social interactions, house training, and sleep wake cycle) was obtained from owners of the dogs by use of a structured telephone interview format. A second interview was performed 12 to 18 months after the initial interview, and differences in responses were evaluated. RESULTS: Sexually intact male dogs were significantly less likely than neutered dogs to progress from mild impairment (i.e., impairment in 1 category) to severe impairment (i.e., impairment in > or = 2 categories) during the time between the first and second interviews. This difference was not attributable to differences in ages of the dogs, duration of follow-up, or the owners' perceptions of the dogs' overall health. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the presence of circulating testosterone in aging sexually intact male dogs may slow the progression of cognitive impairment, at least among dogs that already have signs of mild impairment. Estrogens would be expected to have a similar protective role in sexually intact female dogs; unfortunately, too few sexually intact female dogs were available for inclusion in the study to test this hypothesis. There may be a need to evaluate possible methods for counteracting the effects of loss of sex hormones in gonadectomized dogs. PMID- 11439770 TI - Vertebral scale system to measure heart size in growing puppies. AB - OBJECTIVE: To determine relative heart size in clinically normal puppies and assess whether relative heart size changes with growth. DESIGN: Prospective radiographic study. ANIMALS: 11 puppies without evidence of disease. PROCEDURE: Standardized measurements of the long and short axes of the heart, midthoracic vertebrae, and other structures were made at 3, 6, 12, and 36 months of age. Measurements were recorded in millimeters and number of thoracic vertebral lengths spanned by each dimension, measured caudally from T4 on lateral radiographic views. The long and short axis measurements of the heart, expressed in vertebral lengths, were added to yield vertebral heart size. RESULTS: Mean +/- SD vertebral heart sizes on lateral radiographic views at 3, 6, 12, and 36 months of age were 10.0 +/- 0.5, 9.8 +/- 0.4, 9.9 +/- 0.6, and 10.3 +/- 0.6 vertebrae, respectively. Significant differences were not detected. CONCLUSIONS AND CLINICAL RELEVANCE: Vertebral heart size measurements in puppies are within the reference range for adult dogs (9.7 +/- 0.5 vertebrae) and do not change significantly with growth to 3 years of age. Standards for determining cardiac enlargement are similar in puppies and adult dogs. PMID- 11439771 TI - Extraskeletal osteosarcoma in a cat. AB - A 7-year-old spayed female domestic shorthair cat was referred for evaluation of a localized growing mass on the left flank. Cytologic and histologic findings suggested that the mass was an extraskeletal osteosarcoma. Radiography failed to reveal any association between the lesion and the axial or appendicular skeleton. Because of the large size of the tumor, the cat was treated with carboplatin prior to and after surgery (hemipelvectomy) to ensure that surgical margins were free of neoplastic cells and to prevent systemic dissemination of malignant cells. The tumor has not recurred during a 2-year follow-up period. PMID- 11439772 TI - Osteodystrophia fibrosa in two guinea pigs. AB - Two adult guinea pigs were examined because they were lethargic and reluctant to walk. Additionally, I guinea pig had otitis media, and the other had dental malocclusion. Both guinea pigs had been fed a commercially available diet of cereals and pellets enriched with vitamin C and formulated for this species. Radiographically, the guinea pigs had coarse trabecular bone patterns, skeletal deformations, pathologic fractures, and polyarthritic degenerative joint disease. A double cortical line was also evident on several long bones, the pelvis, and the vertebrae. A diagnosis of osteopenia was confirmed by use of dual-energy x ray absorptiometry. Analysis of a food sample fed to 1 guinea pig revealed calcium and phosphorus contents of 0.524 and 0.425%, respectively (Ca:P ratio, 1.23:1). Microscopic examination of bone tissue from both guinea pigs revealed severe fibrous osteodystrophy. Nutritional secondary hyperparathyroidism caused by calcium-phosphorus imbalance was considered to be the underlying cause of osteodystrophia fibrosa in both guinea pigs. PMID- 11439773 TI - Estimate of the national incidence of and operation-level risk factors for colic among horses in the United States, spring 1998 to spring 1999. AB - OBJECTIVE: To estimate the national incidence of, operation-level risk factors for, and annual economic impact of colic among horses in the United States during 1998 and 1999. DESIGN: Epidemiologic survey. ANIMALS: 21,820 horses on 1,026 horse operations in 28 states. PROCEDURES: Horses were monitored for colic for 1 year, and results were recorded in a log that was collected quarterly. Operation level data were collected via 4 on-site personal interviews. Associations between colic and independent variables adjusted for size of operation were determined. RESULTS: Annual national incidence of colic in the US horse population was estimated to be 4.2 colic events/100 horses per year. Case fatality rate was 11%, and 1.4% of colic events resulted in surgery. Annual cost of colic in the United States was estimated to be $115,300,000. CONCLUSIONS AND CLINICAL RELEVANCE: The national impact of equine colic is substantial because of the high case fatality rate. PMID- 11439774 TI - Congress considers minor use/minor species bill. PMID- 11439775 TI - Risk factors associated with development of postoperative ileus in horses. AB - OBJECTIVE: To determine risk factors associated with development of postoperative ileus in horses undergoing surgery for colic. DESIGN: Case-control study. ANIMALS: 69 horses that developed ileus after surgery for colic and 307 horses that did not develop postoperative ileus. PROCEDURE: Signalment, history, clinicopathologic data, treatment, lesions, and outcome were obtained from medical records. RESULTS: Variables associated with increased risk of postoperative ileus included age > 10 years, Arabian breed, PCV > or = 45%, high serum concentrations of protein and albumin, anesthesia > 2.5 hours' duration, surgery > 2 hours' duration, resection and anastomosis, and lesions in the small intestine. Enterotomy reduced the risk of postoperative ileus. After multivariate logistic regression, the final model included the variables Arabian breed, PCV > or = 45%, lesion type, duration of surgery (> 2 hours vs < or = 2 hours), and pelvic flexure enterotomy. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that by evaluating certain factors, horses at increased risk of postoperative ileus may be recognized before the condition develops. Preventative treatment and early intervention may be instituted in these horses. Shortening surgery time and performing an enterotomy may decrease the probability of horses developing postoperative ileus. PMID- 11439776 TI - Neonatal isoerythrolysis involving the Qc and Db antigens in a foal. AB - In 1992, a multiparous 13-year-old Thoroughbred mare and her 48-hour-old colt were examined because of possible neonatal isoerythrolysis (NI). Supportive treatment was administered, and the foal recovered without requiring a transfusion. According to the owners, the mare had delivered foals without incident during 1987 and 1991. The mare was barren during 1993, but in 1994, delivered a filly that developed severe NI. The foal was given 3 transfusions and eventually recovered without complications. Blood typing analysis of the mare and its foals indicated that all 4 foals were positive for the Qc, Db, and Dq antigens, and the 3 most recently born foals were positive for the Ua antigen; however, the marc was negative for the Qc, Db, Dq, and Ua antigens. The mare did not have alloantibodies against Ua and did not react to the Dq antigen. However, in 1994, the mare reacted against the Db (the reaction was characterized by strong agglutination and an increase in titer at the time of parturition and a subsequent decrease) and Qc (the reaction was characterized by weak lysis and an increase in titer at the time of parturition and a subsequent decrease) antigens. Results of testing in this mare and foals suggested that although approximately 90% of all cases of NI in horses are attributable to the Aa or Qa antigen, other antigens may be involved. PMID- 11439777 TI - Osteitis of the axial border of the proximal sesamoid bones in horses: eight cases (1993-1999). AB - OBJECTIVE: To determine clinical, radiographic, and scintigraphic abnormalities in and outcome of horses with septic or nonseptic osteitis of the axial border of the proximal sesamoid bones. DESIGN: Retrospective study. ANIMALS: 8 horses. PROCEDURE: Data collected from medical records included signalment; history; horse use; severity and duration of lameness; results of perineural anesthesia, radiography, ultrasonography, and scintigraphy; and outcome following surgery. RESULTS: Five horses did not have any evidence of sepsis; the other 3 had sepsis of the metacarpophalangeal or metatarsophalangeal joint or the digital synovial sheath. All horses had a history of chronic unilateral lameness. Three of 5 horses improved after diagnostic anesthesia of the metacarpophalangeal or metatarsophalangeal joint; the other 2 improved only after diagnostic anesthesia of the digital synovial sheath. Nuclear scintigraphy was beneficial in localizing the source of the lameness to the proximal sesamoid bones in 4 horses. Arthroscopy of the palmar or plantar pouch of the joint or of the digital synovial sheath revealed intersesamoidean ligament damage and osteomalacia of the axial border of the proximal sesamoid bones in all horses. All 5 horses without sepsis and 1 horse with sepsis returned to their previous uses. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that osteitis of the axial border of the proximal sesamoid bones is a distinct entity in horses that typically is associated with inflammation of the associated metacarpointersesamoidean or metatarsointersesamoidean ligament and may be a result of sepsis or nonseptic inflammation. Arthroscopic debridement may allow horses without evidence of sepsis to return to their previous level of performance. PMID- 11439778 TI - Age distributions of horses with strangulation of the small intestine by a lipoma or in the epiploic foramen: 46 cases (1994-2000). AB - OBJECTIVE: To test the hypothesis that strangulation of the small intestine by a lipoma or in the epiploic foramen is more common in older horses. DESIGN: Retrospective study. ANIMALS: 46 horses. PROCEDURE: Ages of horses with strangulation of the small intestine by a lipoma (n = 29) or in the epiploic foramen (17) were compared with ages of 79 horses with miscellaneous small intestinal lesions. Effects of increasing age on risk of the diseases of interest were examined by use of logistic regression and a 1-sided trend test for binomial proportions. RESULTS: Mean age of the horses with strangulation in the epiploic foramen (9.6 years) was the same as that for the horses with miscellaneous small intestinal lesions (7.7), but mean age of the horses with strangulation by a lipoma (19.2) was significantly greater than that for the other groups. The proportion of horses with lipoma increased significantly with increasing age, but the proportion with strangulation in the epiploic foramen did not. CONCLUSIONS AND CLINICAL RELEVANCE: Results refute the current suggestion that increasing age predisposes horses for strangulation of the small intestine in the epiploic foramen but support the suggestion that the risk of strangulation of the small intestine by a lipoma increases with age. PMID- 11439779 TI - Interventions for women as family caregivers. AB - Family caregivers are the mainstay of long-term care, as they enable chronically ill elders and children to remain at home. The majority of family caregivers are women and historically their caregiving role has been viewed as an extension of their roles as wife and mother. Although numerous studies report the stresses associated with family caregiving and are predictors of burden, less attention has been given to interventions for family caregivers. The objective of this review is to examine reports of interventions to reduce family caregiver burden, to consider their implications for nursing practice, and to identify directions for future nursing research. PMID- 11439780 TI - Lesbian health and health care. AB - Research on lesbian health and health care is very limited, but is beginning to increase. Evidence of limited access to care, homophobic attitudes of health care professionals, and expected or actual negative experiences in interactions with health care professionals help to explain why lesbians are less likely than other women to seek health care. Lesbians have many of the same physical health needs that other women do, but the most prevalent topics on which research could be found were screening for breast and cervical cancer, sexually transmitted infections (STIs), and HIV. More research has been conducted in areas related to mental health, such as stress, use of therapy, alcohol abuse and recovery, and violence. The chief conclusion from this review is that there is a need for all types of research in all areas of lesbian health. PMID- 11439781 TI - Immigrant women and their health. AB - Immigrant women's health is a relatively new research area. At the beginning of the 1990s, nurse scholars concluded that there was insufficient research on this topic. They recommended broadening the overly narrow research foci on immigrant women's childbearing and on select populations, developing national data bases, identifying high-risk groups, and developing population-specific interventions. This chapter reviews 292 research articles published in journals during the 1990s about adult immigrant women's health. It: (1) summarizes research findings on topics that were the major foci of research conducted in the 1990s, (2) evaluates progress over the last decade in the research agenda proposed above, and (3) makes recommendations for research in the new millennium. PMID- 11439782 TI - Women and stress. AB - This review focused on published research reports that explored the association between stress and women's health and illness. Because of the vastness of the literature, the review was limited to three conceptual areas. The first included those studying the association between stress and illness. Most were correlational in design, thus lacking the power to support assertions of a cause and-effect relationship. The second focused on studies of the psychophysiology of stress. These studies provide nurse scientists and clinicians with insights into the underlying pathology of stress. When paired with behavioral measures, these studies provide a window into brain mechanisms involved in mind/body interactions. The third conceptual area included those studies of stress reduction therapies to influence health and illness. These studies have yielded a mighty arsenal of interventions that may be used to reduce distress. PMID- 11439783 TI - Sleep and fatigue. AB - This chapter provides a review and synthesis of research on women's sleep and fatigue from a nursing perspective. Most of the research involves four primary issues for women: menstrual cycles, childbearing, chronic mental or physical illness, and oncology. Research with healthy women focused on diurnal fluctuations in fatigue and relationships to sleep, without regard for exercise or level of daytime activity. Research on chronic illness and cancer fatigue focused on general fatigue and its impact on activity, without regard for sleep or therapeutic use of rest and naps. A comparison of these two areas highlights gaps in nursing knowledge about sleep and fatigue. Further research is needed to understand relationships between nonrestorative sleep, fatigue, and symptoms related to poor quality of life. From a synthesis of these studies, nonpharmacologic interventions that could be prove useful in promoting a higher quality of life for those with either acute or chronic fatigue are then proposed. PMID- 11439784 TI - Intimate partner violence against women. AB - Intimate partner violence against women has received considerable attention from nurse-researchers over the past 10 years. Although the amount and sophistication of both quantitative and qualitative research have changed over time, nursing research on intimate partner violence against women has not lost its perspective; nurse-researchers have continued to address women survivors' full range of human responses to violence. Research into violence during pregnancy and battered women's psychological responses to abuse have received considerable attention. Research into violence during pregnancy and battered women's psychological responses to abuse have received considerable attention. Research into violence during pregnancy accounts for fully 20% of all the reviewed nursing research. The largely qualitative research into women's psychological responses to violence is particularly rich and remarkably similar across multiple studies. International studies on intimate partner violence are beginning to appear in the literature and research that addresses the unique experience of ethnically diverse women is occurring with greater frequency. The purpose of this chapter is to review nursing research on intimate partner violence against women in the past decade. Future directions for nursing research, practice, and education are included. PMID- 11439785 TI - Conceptual models for women's health research: reclaiming menopause as an exemplar of nursing's contributions to feminist scholarship. AB - An examination of women's transition through menopause provides a remarkable example of nursing's contributions to feminist scholarship. The predominant biomedical model perpetuates the idea that menopause is a deficiency disease, whereas feminist and nurse scholars have deconstructed this paradigm and have reclaimed menopause as a part of midlife women's developmental stage. We begin this chapter with a review of the birth of women's health scholarship as it is the foundation for theory that undergirds feminist nursing research. We then discuss the tenets of feminist scholarship. The historical context of menopause is reviewed briefly to highlight the ways in which menopause was transformed from a normal physiological event to a disease. Using this as a backdrop, we reviewed nursing studies in two emerging bodies of knowledge. We reviewed 10 studies in the area of "women reclaiming menopause" and found that over all women believe the menopausal transition is a normal developmental stage. The second area of new research looks at "menopause across cultures." The studies of Korean, Indian, and Thai women reviewed demonstrate that similar to other health issues, the experience of Western women cannot be universalized, and most important, researchers must take into consideration the social, political, economic, and cultural forces that impact women's experience of the menopause transition. PMID- 11439786 TI - What we know and how we know it: contributions from nursing to women's health research and scholarship. AB - In this first chapter, we trace the historical roots of nursing research and scholarship focused on protecting and promoting women's health. Beginning with Florence Nightingale, modern nursing's first researcher, who focused on the health impact of women's daily lives through her detailed observations of human behavior. More recently, nursing's contributions to women's health over the past 30 years have redefined women's health, proposed new frameworks for understanding women's health; provided reviews of the women's health literature across disciplines; developed communities of nurse scholars and researchers focused on new areas of women's health research; generated and expanded the knowledge base for women's health practice and education; promoted a global view of women's health; and proposed new models for women's health care delivery. Clearly, a community of nursing scholars, developed over the past 25 years, has contributed to advancing women's health knowledge and improving the health and well-being of women. Without the benefit of a crystal ball, we suggest that nursing will continue to provide leadership in the conduct and the application of research to improve women's health and women's lives. PMID- 11439787 TI - Health decisions and decision support for women. AB - Women are more likely to live longer with chronic illness and have a long-term relationship with their health care provider; this requires a situation in which patients and providers have a role in managing illness. In this chapter, the authors provide a conceptual overview of decision making along with key issues: historical concepts related to patients and providers, consumerism, informed choice/consent, patient rights, shared decision making, patient involvement, as well as an overview of models of patient/provider partnerships. This review builds on the work of O'Connor et al. (1999), which resulted in a Cochrane review of decision aids and focuses the examination of patient decision aids that support women's decisions regarding health treatment or screening. The authors conclude with a look to the future and recommendations for research in the area of shared decision making and health care decision aids. PMID- 11439788 TI - Female troubles: an analysis of menstrual cycle research in the NINR portfolio as a model for science development in women's health. AB - The National Institute for Nursing Research (NINR) has been active in developing a research portfolio of investigator-initiated studies in addressing the cause and consequences of menstrual cycle and menopause-related health problems. This chapter provides an overview of the nature and level of research activity funded by NINR since its inception in 1986, major findings generated by the most successful award recipients, the impact on the broader field of women's reproductive health and directions for future research. Presented here is an analysis of research designs and methodologies framed within the context of 4 stages of scientific development in the field: exploratory, descriptive studies in well women; illness as a biobehavioral phenomenon; knowledge generation in understudied populations; and the development and testing of clinical therapeutics for symptom management and health promotion strategies. Nursing science contributions to the NINR portfolio of women's health research has been focused primarily on the definition and management of the symptoms of premenstrual syndrome and menopause. The increasing numbers of intervention studies suggests a coming-of-age in nursing science with respect to the development of evidence-based outcome data for the management of menstrual cycle and menopause-related symptoms. Clearly, the range and diversity of NINR grant funded activity suggest that menstrual cycle research is a strong area of interest in nursing science. PMID- 11439789 TI - Women as mothers and grandmothers. AB - This chapter analyzes the literature on women as mothers; research reports published between January 1985 and December 1999 were reviewed. As in the past, almost all of the extant studies analyzed the experience of mothers in their children's first year of life. Although therapeutic suggestions were made in many studies, relatively few interventions have been implemented and evaluated. More studies are needed that go beyond traditional family forms and that explore mothers' role development over the full course of their children's growth and development. Additional longitudinal research that views maternal role development as a process is indicated. PMID- 11439790 TI - Women and employment: a decade review. AB - Contemporary women fulfill multiple social roles: wife or partner, parent and caregiver to elders, worker in the labor force. This chapter focuses on women and employment. Nursing research from the past decade on women and their role in the work force, with emphasis on the relationships between paid work and women's other social roles and their health is reviewed. Major categories of nursing research contributions are summarized, including populations studied, methodological approaches, and major findings. Suggestions for emphases in future research are included. PMID- 11439791 TI - The history of intravenous anaesthesia: the barbiturates. Part 2. PMID- 11439792 TI - Complementary medicine in intensive care: ethical and legal perspectives. AB - Complementary medicine continues to increase in popularity in the general community. As a result it is likely that requests for the administration of complementary medicine to intensive care patients will be more frequent in the future. It is therefore prudent for intensive care clinicians to address this issue and develop an approach that is consistent. Complementary medicine has not been subjected to well conducted trials to determine its efficacy and risks. Consequently decisions about its use cannot be based on risk/benefit analyses and genuine informed consent cannot be achieved. Therefore complementary medicine should not be incorporated into intensive care practice. Strict adherence to a policy of negating requests for administration of complementary medicine in intensive care patients may result in significant conflicts between intensive care clinicians, patients and families. On occasions the patient or family may insist on the use of complementary medicine and it may be seen as important to their psychological wellbeing to accede to the request. The intensive care clinician is still legally responsible for any treatment administered to the patient, even if it is against medical advice. Nevertheless if there is no demonstrable risk to the patient, complementary medicine can be administered following appropriate counselling and documentation. This review addresses the legal and ethical difficulties that may arise and an approach that may be followed when requests are made for complementary medicine in intensive care patients. PMID- 11439793 TI - Do anaesthetized patients recover better after Bispectral Index Monitoring? AB - Could Bispectral Index (BIS) monitoring during anaesthesia improve the recovery characteristics of patients? Previous studies have shown conflicting results. To eliminate bias, a control group of 75 cases anaesthetized by the authors was compared to a reference group of 141 cases anaesthetized by other anaesthetists. A study group of 71 cases was then anaesthetized by the authors titrating to BIS 40-50 and this was compared with the control group. Recovery characteristics were assessed using a Modified Aldrete score. BIS monitored cases had improvements in blood pressure stability (P = 0.023) and respiratory score (P = 0.016) throughout the study period. Activity and consciousness levels were higher on arrival in PACU in the BIS monitored group (P = 0.015 and P = 0.017) but were not maintained. There was no significant difference in mean oxygen saturation scores. The improved recovery characteristics in BIS monitored patients may have positive implications for safety, nursing workload, staffing, and cost savings in the PACU. PMID- 11439794 TI - PCA ketamine and morphine after abdominal hysterectomy. AB - Following a standardized general anaesthetic for total abdominal hysterectomy, patients received either patient controlled analgesia (PCA) with morphine 1 mg/ml (group M, n = 33) or morphine 1 mg/ml plus ketamine 2 mg/ml (group K, n = 37) for 48 hours in a randomized, double-blind fashion. In 43 women the area of allodynia around the scar was mapped as a measure of the degree of central sensitization. A significant reduction in the area of allodynia was found in those receiving ketamine with morphine (42 cm2 [interquartile range (IQR) 57] compared with 57 cm2 [IQR 82] z = -2.0, P = 0.04) in those receiving morphine alone. There were no significant differences between the two groups with respect to age, or weight, or between the subgroups within which the area of allodynia was measured with respect to length of incision. No significant differences were found between the groups with respect to pain scores, total or hourly drug consumption, patient satisfaction, nausea scores or antiemetic use. Patients in group K were more likely to require PCA for a shorter period than those in group M (median 40 hours, IQR 26 versus 48 hours IQR 7). Ten patients in group K were withdrawn because of side-effects (dysphoria n = 4, nausea n = 2, pruritus n = 4) compared with one in group M (nausea n = 1) (P = 0.006). The potential usefulness of ketamine after hysterectomy was offset by a high incidence of adverse effects and a lack of opioid-sparing effects, such that combined intravenous ketamine and morphine PCA as used in this study cannot be recommended for routine care. PMID- 11439795 TI - Blind orotracheal intubation with the intubating laryngeal mask versus fibreoptic guided orotracheal intubation with the Ovassapian airway. A pilot study of awake patients. AB - In a randomized, prospective pilot study, we compared awake blind orotracheal intubation using the intubating laryngeal mask airway (blind-ILM) with awake fibreoptic-guided orotracheal intubation using an Ovassapian airway (FOS-OA). Fifty-four patients (ASA 1 to 3, aged 18 to 85 years) requiring awake intubation for elective surgery were randomly allocated by coin toss into two groups: 31 patients were intubated blindly through the ILM (blind-ILM) and 23 were intubated using fibreoptic guidance through the Ovassapian airway (FOS-OA). Sedation to a target clinical end-point (spontaneous eye-closing, but responsive to verbal command) was obtained with fentanyl/midazolam and a cricothyroid puncture was performed with 3 ml lignocaine 4%. The oropharynx was then topicalized until tolerance of a Guedel airway was achieved. The number of failed attempts (maximum of three allowed), overall success rates, the time from insertion of the airway to capnographic (blind-ILM) or fibreoptic (FOS-OA) confirmation of intubation or until three failed attempts, and cardiovascular responses before and during intubation, were recorded. The first time (blind-ILM, 25/31 [81%]; FOS-OA, 20/23 [87%], P = 0.6) and overall (blind-ILM, 26/31 [84%]; FOS-OA, 22/23 [96%], P = 0.2) intubation success rates were similar. The mean +/- SD time to intubation was shorter for the blind-ILM group (104 +/- 65 vs 158 +/- 115 sec, P = 0.05). There were no clinically significant differences in blood pressure or heart rate between groups. Compared with baseline values, there was no cardiovascular response to intubation in either group. We conclude that the blind-ILM and FOS-OA techniques have similar success rates and cardiovascular responses, but intubation is slightly quicker with the blind-ILM technique. PMID- 11439796 TI - Intubation through intubating laryngeal mask with and without a lightwand: a randomized comparison. AB - The combined use of a lightwand and the intubating laryngeal mask airway (ILMA) was compared with the use of the ILMA alone to determine whether the combination was a more efficient method of endotracheal intubation. One hundred healthy patients were randomly assigned to two groups. After induction of anaesthesia, Group A patients were intubated blindly through the ILMA while in Group B, intubation was guided by a lightwand. A sequence of standard manoeuvres was followed it attempts at intubation failed. The number of manoeuvres used, the time taken for successful intubation and complications associated with intubation were recorded. Intubations were successful in all patients, but the mean endotracheal intubation time was longer in Group A than in Group B (38.3 +/- 10.4 s versus 26.4 +/- 9.1 s, P < 0.001). The number of patients who needed one or more manoeuvres was significantly higher in Group A than in Group B (76% versus 42%, P = 0.001). We conclude that the lightwand is a useful adjunct in endotracheal intubation through an ILMA. PMID- 11439797 TI - Performance of transcutaneous PCO2 and pulse oximetry monitors in newborns and infants after cardiac surgery. AB - We examined the effect of core and skin temperature on the accuracy of two pulse oximeters (Nellcor Symphony and Hewlett Packard saturation module, M1020A) and a transcutaneous PCO2 monitor (Fastrac Transcutaneous monitor) immediately after cardiac surgery in a group of newborns and infants. Seventy-nine sets of data were collected from 46 patients. Core temperatures ranged from 35.3 degrees C to 39.4 degrees C, skin temperatures ranged from 27.0 degrees C to 37.4 degrees C and core-skin temperature gradients ranged from 0.1 degree C to 10.1 degrees C. Data analysis consisted of comparing the difference between transcutaneous PCO2 and arterial PCO2 and the differences between oxygen haemoglobin saturation measured by both pulse oximeters and oxygen haemoglobin saturation measured by co oximeter to core temperature, skin temperature and core-skin temperature gradients. The mean differences +/- standard deviations and limits of agreement for transcutaneous PCO2 and oxygen haemoglobin saturation measured by the Hewlett Packard and Nellcor pulse oximeters were 0.95 +/- 4.10 mmHg (-7.09 mmHg to 8.99 mmHg), -1.07 +/- 1.84% (-4.68% to 2.54%) and -1.23 +/- 2.23% (-5.60% to 3.14%) respectively. Analysis of correlation coefficients showed that the accuracy of the transcutaneous PCO2 monitor and the pulse oximeters were not affected by core temperature, skin temperature or core-skin temperature gradient in the ranges encountered. We therefore conclude that these devices are acceptably accurate and suitable for use in infants when core and skin temperatures and core-skin temperature gradient are in the range normally found after cardiac surgery. PMID- 11439798 TI - Management of postoperative epidural analgesia: a survey of Australian practice. AB - A sample of Australian hospitals was surveyed about their practice preferences in relation to the management of epidural analgesia for postoperative pain. Results indicated substantial variation in practice preference across institutions with respect to observation protocols, epidural analgesia duration, catheter removal where anticoagulant therapy is concurrent and management of catheter problems. Further research appears necessary to develop optimal epidural analgesia management practice. PMID- 11439799 TI - Ventilator-patient dyssynchrony induced by change in ventilation mode. AB - Patient-ventilator interactions may be coordinated (synchronous) or uncoordinated (dyssynchronous). Ventilator-patient dyssynchrony increases the work of breathing by imposing a respiratory muscle workload. Respiratory centre output responds to feedback from respiratory muscle loading. Mismatching of respiratory centre output and mechanical assistance results in dyssynchrony. We describe a case of severe patient-ventilator dyssynchrony and hypothesize that dyssynchrony was induced by a change in mode of ventilation from pressure-cycled to volume-cycled ventilation, due to both ventilator settings and by the patient's own respiratory centre adaptation to mechanical ventilation. The causes, management and clinical implications of dyssynchrony are discussed. PMID- 11439800 TI - Retro-orbital tumour--an uncommon cause of headache in pregnancy. AB - Retro-orbital tumour was the cause of headache and neuropathic facial pain in a 31-year-old pregnant woman. The diagnosis had been overlooked as a result of a long history of migraine. There was exacerbation of the pain throughout the pregnancy, particularly in the third trimester. Pharmacological agents commonly used to manage neuropathic pain states were relatively contraindicated due to potential adverse effects on the fetus. Cognisant of such limitations imposed by pregnancy, we used multimodal therapy in an attempt to control the pain. This included morphine, paracetamol, amitriptyline, ketamine and psychological support. The management challenges are described. PMID- 11439801 TI - Severe convulsant hypomagnesaemia and short bowel syndrome. AB - Hypomagnesaemia as a primary cause of a generalized seizure is uncommon. A 60 year-old woman with Crohn's disease, who had had recent small bowel surgery and a total colectomy 10 years previously, was admitted complaining of severe nausea, vomiting, fatigue and thirst. Despite oral magnesium therapy she had a generalized seizure due to severe hypomagnesaemia of 0.09 mmol/l (normal range 0.65 to 1.05 mmol/l). Her serum calcium was 1.91 mmol/l (2.03 to 2.63 mmol/l). Hypomagnesaemia can cause generalized convulsions but is usually associated with hypocalcaemia. This patient had an almost normal serum calcium level, and therefore hypomagnesaemia would seem to have been the direct cause of her seizure. Long-term intravenous magnesium was necessary to prevent further seizures. PMID- 11439802 TI - Intraoperative fibreoptic bronchoscopy during neonatal tracheo-oesophageal fistula ligation and oesophageal atresia repair. AB - Maintenance of adequate ventilation under anaesthesia can be difficult during identification and ligation of congenital tracheo-oesophageal fistula with repair of oesophageal atresia. Anaesthesia may also be complicated by problems associated with prematurity, pre-existing aspiration pneumonitis, and difficulty positioning the endotracheal tube to prevent inflation of the stomach with increased risk of aspiration and diaphragmatic splinting. Even intubation of the fistula and gastric rupture may occur. Two neonatal cases are presented where use of a 2.2 mm neonatal bronchoscope passed through a 3.0 mm ID tracheal tube facilitated surgical identification of the fistula, diagnosis of fistula intubation and other airway problems intraoperatively. PMID- 11439803 TI - Airway protection with the ProSeal laryngeal mask airway. AB - The ProSeal laryngeal mask airway is a new laryngeal mask device that has a modified cuff and a drainage tube. We describe a case where the ProSeal laryngeal mask successfully channelled regurgitated fluid away from the respiratory tract. PMID- 11439804 TI - Missed diaphragm rupture following blunt trauma. AB - Traumatic rupture of the diaphragm resulting from blunt trauma remains a challenging clinical entity. Diagnosis remains difficult and is often delayed. A prompt diagnosis requires a high index of suspicion. We describe two patients with ruptured diaphragms in whom the diagnosis was made at different stages of their illness. The diaphragmatic injury reflects the magnitude of their associated injuries and probably contributes to the overall mortality. PMID- 11439805 TI - Severe adult botulism. AB - A case of severe adult botulism with paralysis, respiratory failure and cranial nerve palsies is presented. The pathophysiology, clinical manifestations, diagnosis and treatment options for botulism are discussed. PMID- 11439806 TI - Fire during thoracic surgery. AB - A 67-year-old male with bilateral lung lesions presented for median sternotomy. One-lung ventilation was complicated by arterial desaturation. Continuous positive airway pressure with oxygen was applied to the non-ventilated lung to relieve the hypoxaemia. However, the cotton gauze packed inside the operative site was ignited by the electrocautery. The burning gauze was immediately removed without any complication. We report a case of electrocautery-induced fire during thoracic surgery done in supine position and discuss its implication on the choice of oxygen enrichment therapy during one-lung ventilation. PMID- 11439807 TI - Potential problems with BLUELINE ULTRA tracheostomy tube with inner cannula. PMID- 11439808 TI - Estimating the diameter of the left main bronchus is a clinically useful method for selecting left double-lumen tubes. PMID- 11439809 TI - Which should be inserted first--giving set or airway? PMID- 11439810 TI - Intravenous equipment--infusions. PMID- 11439811 TI - About intrathecal baclofen in tetanus. PMID- 11439812 TI - A hierarchical analysis of long-term illness and mortality in socially deprived areas. AB - This article is a multilevel analysis of the effects on self-reported long-term illness and mortality of the socio-economic position of the neighbourhood. Using data from the Swedish Level of Living Survey, neighbourhood social position is measured by a composite Care Need Index, (CNI) together with such indicators of individual socio-economic position as occupation and housing tenure, with adjustment for age, sex, marital status and social network. Data came from 22,236 people aged 25-74, and were collected from 1988 to 1992. The cross-sectional data were analysed using a hierarchical logistic regression model. In a second analysis, each participant was followed from the initial interview until his or her death, or until the termination of data collection (31 December 1996). A neighbourhood's low social position and an individual resident's low socio economic position (i.e., a manual worker, or person renting a flat) were found to be associated with increased risk of long-term illness. We conclude that a neighbourhood's low socio-economic position, that is, a high score on the CNI, is a risk factor for long-standing illness above and beyond an individual's socio economic position. The differences in mortality could be explained by the included independent individual variables. PMID- 11439813 TI - Children's behaviour and the urban environment: an ecological analysis. AB - This research examines the spatial distribution and ecological correlates of problem behaviour amongst children living in the most highly urbanised portion of Greater Victoria, British Columbia, Canada. Rates of problem behaviour within the sample differ substantially, at the census tract level, with respect to the mean rate calculated for the study area as a whole. While the degree of spatial concentration varies between behaviour types, each exhibits a similar overall trend, with the highest rates in the western and/or central portions of the study area, and the lowest rates around its northern and eastern periphery. Bivariate correlation analyses reveal significant ecological relationships between rates of problem behaviour and census measures of aggregate socio-economic status. The degree to which the urban ecology of the study area is related to census tract rates of problem behaviour is dependent upon the nature of the behaviour being considered. Results of contextual analyses using five ecological variables and their family-level equivalents suggest that, in some cases, census tract characteristics may have an independent impact upon behavioural status. PMID- 11439814 TI - Do high levels of unemployment influence the health of those who are not unemployed? A gendered comparison of young men and women during boom and recession. AB - Research has shown that health among young people, particularly women, deteriorates during a recession compared to a boom. It seems that the trade cycle mainly affects the health of those who are not long-term unemployed. The aim of this study was to analyse the relation between the health of non-unemployed people and the levels of unemployment in society. Two groups of young people aged 21 were surveyed, one in 1986 (the boom group, n = 1083) and one in 1994 (the recession group, n = 898). The non-response rate was 2% in the first and 10% in the second group. Both groups were investigated with a self-administered questionnaire, which included questions about somatic and psychological health, as well as experiences of employment, unemployment, education and labour market programmes. Young men and women generally reported more somatic and psychological symptoms during recession than boom. The only exception was psychological symptoms among men, which was of the same magnitude during both periods. Poorer health during recession was found among women in work and in labour market programmes, as well as among both male and female students. Multiple regression analysis was performed in order to analyse if the occupational-related health effects of the trade cycle remained after controlling for possible moderating factors. The effects of unemployment in society on young people's health may be mediated through pessimism about the future, high demands and financial problems. Lack of control over the work situation may also be an important contributing factor to ill health among women during recession. The trade cycle was correlated with ill health among women only. Possible explanations for poorer health among women during a recession were discussed. PMID- 11439815 TI - Unemployment and psychosocial adjustment in young adults: causation or selection? AB - This study investigates the relationships between unemployment following school leaving and psychosocial adjustment problems (mental health, substance use, crime, suicidal behaviours and teenage pregnancy) in a birth cohort of over 1000 New Zealand born young people. The data were gathered during the course of the Christchurch Health and Development Study (CHDS). The CHDS is a longitudinal study of a birth cohort of 1265 children born in the Christchurch (NZ) urban region who have been studied from birth to age 21. Data were gathered by personal interview on: (a) exposure to unemployment and (b) personal adjustment over the period from age 16 to age 21. Measures of personal adjustment included mental health (depression, anxiety), substance use, crime, suicidal behaviours and (for females) teenage pregnancy. Data were analysed using a fixed effects regression model that took into account both observed and non-observed sources of confounding and the possibility of reverse causal associations between personal adjustment and unemployment. Before adjustment for confounding and reverse causality there were significant (p < 0.001) associations between exposure to unemployment and measures of mental health, substance use, crime, suicidal behaviours and teenage pregnancy. Adjustment for confounding factors and reverse causality reduced these associations quite substantially and after control for sources of confounding a number of associations became nonsignificant. Nonetheless, after such control, exposure to unemployment remained significantly (p < 0.05) associated with suicidal ideation, substance abuse and criminal behaviours. It is concluded that, in part, the associations between unemployment and personal adjustment are spurious and reflect the presence of confounding factors that are related to both unemployment and adjustment. Nonetheless, the findings suggest that exposure to unemployment may be associated with increased risks of suicidal thoughts, crime and substance use. PMID- 11439816 TI - Health psychology and the study of the case: from method to analytic concern. AB - This paper recommends that the study of the case be seen as of primary analytic concern to social scientists, and particularly to health psychologists. Criticism is made of the idea that a case is merely a medical instance or a methodological option. Instead, we argue that health psychologists should re-direct their attention to the 'study of the case' as being central to issues concerning health, illness and healing. There are three reasons for doing this. First, case study is basic to any procedure that involves collecting information about the context in which medicine is practiced. Second, communication between health professionals involves presenting the clinical situation of their patients as storied accounts, so that cases are made, not found. Third, there is presentational work by patients, involving communications of (as well as about) suffering and relief. This last feature is basic to how doctors and health psychologists, especially those engaged in clinical work, understand individuals as 'cases'. The paper explores differences between these different forms of case, while emphasizing portrayal as a key feature of all of them. In justifying the study of the case on conceptual as well as on clinical and methodological grounds, we highlight the position of health psychology in its attempts both to study and to intervene in health-care contexts. PMID- 11439817 TI - Evaluation of medical rehabilitation in community based rehabilitation. AB - Almost all governments and non-governmental organisations in developing countries use a community-based rehabilitation (CBR) approach to work with disabled people. Although disabled people's organisations reject the categorisation of disability in individual terms, 'medical rehabilitation' is still regarded as an important but time limited process within rehabilitation. The paper lists measures and methods used in a comprehensive evaluation, and presents a practical method to examine the quality of medical rehabilitation. The method was developed and applied in an evaluation of service needs and service provision for disabled people in low-income communities, for the Ministry of Welfare, Government of India. The method described is a tracer approach. It assesses quality in three aspects of medical rehabilitation: (i) Technical quality, based on application of minimum technical standards for each impairment. (ii) Interpersonal quality, by observation of service sessions and interviews with service users. (iii) Management (structural) quality, by comparing the rehabilitation goals of service users and service providers. The method differs from most others in that it is process oriented, as opposed to output oriented. The method meets the challenges of providing low-cost assessment of a difficult outcome measure (the quality of medical rehabilitation), within a complex process (CBR). It is anticipated that the tracer method will be useful to the objective evaluation of disability services throughout the developing world. PMID- 11439818 TI - British psychiatry in the 20th century--observations from a psychiatric journal. AB - In order to investigate change and continuity in the concerns and practices of psychiatry in Britain during the 20th century we examined contents of the British Journal of Psychiatry. Specifically we sought to examine the paradigms used by psychiatry to conceptualize mental illness during this time. Back issues of the journal for 1 year at the mid-point of each decade were examined. We undertook a quantitative analysis categorising each article in terms of its form and content and a qualitative analysis in order to summarise the subjects that were covered. The results show that there has been continuous interest in biological aspects and treatments of mental illness with relatively little coverage of psychoanalysis or social psychiatry. Little support was found for the suggestion that major shifts have occurred in the explanatory paradigms used by psychiatry over the century. Modern interest in biological psychiatry is therefore not a new departure, but appears rather as the continuation of a long-standing inclination. The decline of the asylum-based system of care has been accompanied by a broadening in the scope of psychiatric concerns with a greater emphasis on milder mental disorders such as neurosis and depression. PMID- 11439819 TI - Physician-patient communication about over-the-counter medications. AB - The purpose of this study was to describe physician-patient communication about over-the-counter medications using a data set comprised of audio-tapes and transcripts of 414 primary care medical visits. The data set was collected during 1995 at the family practice and general medicine clinics at the University of New Mexico Health Sciences Center. Twenty-seven resident physicians and 414 of their adult patients participated. Fifty-seven percent of patients reported using one or more OTC medications during the past month. Analgesics, cold or allergy products, and antacids were the most commonly used OTC medications. White patients were significantly more likely to have reported using an analgesic in the past month than non-white patients. Female, white, and younger patients were more likely to have reported using a cold or allergy product in the past month than male, non-white, and older patients. Approximately 58% of patients discussed OTC medications with their physicians. Older patients and female patients as well as patients who reported using an antacid in the past month were significantly more likely to have discussed OTC medications with their physicians. Physicians asked questions about OTC medications during only 37% of encounters. Patients asked questions about OTC medications during 11% of encounters. Patient ethnicity did not influence physician or patient question-asking and information-giving about OTC medications. Male physicians were more likely to state information and ask questions about OTC medications than female physicians. Patients were more likely to ask male physicians questions about OTC medications. Physicians were more likely to state OTC information to and ask OTC questions of female and older patients. Physicians were more likely to ask less educated patients questions about OTC medications. Less educated patients were more likely to ask physicians questions about OTC medications. Despite the fact that more than half of all patients reported using OTC medications, physicians asked questions about OTC use during only approximately one-third of encounters. Of patients who reported using an OTC medication in the past month, 58% did not tell their physicians, yet only 14% of patients believed that it was not important for the physician to know about their OTC use. Physician-patient communication about OTC medications should be encouraged so that the patient becomes a collaborative partner in medication management. PMID- 11439820 TI - Sexual cleansing (Kusalazya) and levirate marriage (Kunjilila mung'anda) in the era of AIDS: changes in perceptions and practices in Zambia. AB - Since sexual cleansing (kusalazya) and the intertwined ritual of levirate marriage or widow and widower inheritance (kunjilila mung'anda) have come to be implicated in the transmission of HIV/AIDS, alternative rituals to sexual cleansing have emerged. Using both quantitative and qualitative data obtained from Zambia in the second half of 1998, this study reveals that the alternative rituals to sexual cleansing include sliding over a half-naked person (kucuta) or over an animal (kucuta ng'ombe or cow-jumping); use of herbs and roots (misamu); cleansing by a married couple. Concoctions or other rituals that were otherwise considered 'alien' in Southern Province, such as cutting of hair (kugela masusu) and application of some powder (kunanika busu), have also been adopted. The study, therefore, discusses various aspects of these alternative practices: who performs them and how; whether the processes are connected to polygyny (maali), levirate marriage (kunjilila mung'anda), and grabbing or inheriting property (kukona); and whether these practices are also risk factors in the spread of HIV/AIDS. PMID- 11439821 TI - Determinants of the intention of Dutch adolescents to register as organ donors. AB - This article identifies relevant predictors of willingness among Dutch adolescents to register as posthumous organ and tissue donors. A cross-sectional survey was conducted among 1836 Dutch adolescents between 16 and 18 years of age who had not yet been approached for registration. Sixty-seven percent of the participants indicated that they were willing to register as organ donors. In order of greatest association, negative outcome beliefs, anxiety, social outcome expectations, involvement with organ donation issues, positive outcome beliefs, knowledge about organ donation registration, past behaviour, and sex were significant predictors of the willingness to register as organ donor. Religion and level of education had a significant bi-variate correlation with intention, but were not significant predictors after controlling for other variables. The results give clear suggestions for tailoring organ donation registration interventions to specific characteristics, beliefs and misconceptions of adolescents. PMID- 11439822 TI - Sex selection in practice among Hong Kong Chinese. AB - The aim of this study is to identify factor(s) influencing imbalance in the sex distribution of children (more children of the same sex) of the multiparous women attending an antenatal clinic in Hong Kong. A retrospective study was carried out to compare the expected and observed sex ratios of babies born to women with one previous live birth and women with two. All singleton pregnant women of parities 1 and 2 delivering after 22 weeks of gestation, between 1 July 1996 and 30 June 1998, were included. A total of 2604 women of parity 1 and 752 women of parity 2 were included. The sexes of the children of parity 1 women and the sex distribution of their newborns were consistent with the predicted value. The parity 2 women were more likely to have third children if their previous children were of the same sex (p-value < 0.001). This implies that the parity 2 women attending our antenatal clinic had third pregnancies because of social, economical or environmental reasons. Our parity 2 women with two daughters were more likely to have sons (p < 0.05). Thus, biological or parental factors were unlikely to be important in influencing the sex of the newborn. This suggested that sex selection or sex-selective abortion might be practiced among Hong Kong Chinese women. PMID- 11439823 TI - Investing in children, families, and communities: challenges for an interdivisional public policy collaboration. AB - An interdivisional collaboration to foster the development of strengths-based policies for children, youth, families, and communities is described. The initiative includes (1) producing a book that integrates scholarly research and policy; (2) developing materials for policymakers to use, including a policy oriented summary of the book; (3) enhancing the capacities of the divisions to communicate with and influence policymakers; and (4) taking action steps to influence policymakers. During the process of developing these products, a number of tensions emerged between the academically-based and policy-based authors of the book, many of which centered around how the information should be presented and, in particular, how to formulate and present policy recommendations. Tensions fell into four general categories: understanding the appropriate scope for recommendations, using the right language, understanding the kind of information that is needed, and understanding the bottom line. The author concludes by urging psychologists to become adept at understanding and participating in the public policymaking process. PMID- 11439824 TI - The benefits of mutual support groups for parents of children with disabilities. AB - This study examined what parents of children with disabilities and special needs found helpful about belonging to mutual support groups. Quantitative data (based on 56 parents from 6 groups) indicated that members found the groups very helpful and were very satisfied with the support they received from their groups; they also described the groups as high in cohesion, expressiveness, task orientation, and self-discovery. A grounded theory analysis of focus group data (based on 43 parents from 5 of the groups) indicated that such support was helpful in three broad domains: (1) the sociopolitical, which involved developing a sense of control and agency in the outside world; (2) the interpersonal, which involved a sense of belonging to a community; and (3) the intraindividual, which involved self change. A central theme of identity change emerged as superordinate to these three categories. PMID- 11439825 TI - The coming-out process and its adaptational and health-related associations among gay, lesbian, and bisexual youths: stipulation and exploration of a model. AB - A model is proposed and explored that links the coming-out process to the psychological functioning (i.e., self-esteem and distress) and sexual behaviors of gay, lesbian, and bisexual youths recruited from gay-focused community-based and college organizations in New York City. The coming-out process is multidimensional, consisting, as defined here, of involvement in gay/lesbian activities, attitudes toward homosexuality, comfort with homosexuality, self disclosure of sexual identity to others, and sexual identity. The coming-out dimensions were related to self-esteem, distress, and unprotected sexual behaviors. In addition, the relations between the coming-out dimensions and unprotected sexual behaviors were explained by psychological functioning. In particular, limited involvement in gay/lesbian activities was associated with more unprotected sex. Negative attitudes toward homosexuality were related directly to more unprotected sex, and they were related indirectly to more unprotected sex by means of increasing emotional distress. These and other findings have implications for designing preventive interventions to increase the youths' psychological functioning and reduce their unprotected sexual behaviors. PMID- 11439826 TI - The intersection of, and interaction between, theory development and practice in prevention science: Irwin Sandler, Outstanding Contributions to Theory development. PMID- 11439827 TI - Quality and ecology of adversity as common mechanisms of risk and resilience. AB - This paper proposes common mechanisms to explain the effects of adversities and of resources that promote resilience. Adversities threaten the satisfaction of basic human needs and the acquisitions of competencies to carry out valued social roles. Adversities can also be characterized in terms of their ecological properties of occurrence in time, and place. Resilience resources at the individual, microsystem and macro levels reduce the negative effects of adversities through their effects on satisfaction of basic human needs and their effects on the occurrence of adversities. The effects of resilience resources are described as preventive, protective and promotive. Implications are presented for the development of interventions to promote resilience. PMID- 11439828 TI - Introduction of Stephanie Riger, the 2000 Recipient of the Society for Community Research and Action Award for Distinguished Contributions to Theory and Research. PMID- 11439829 TI - Transforming community psychology. AB - Community psychology has made great strides in including context when understanding people in their environments. While continuing to consider context, we need to expand our conceptualization of the individual in community settings. I propose 3 principles: (1) focus our research on people, not programs; (2) consider multiple dimensions of people's experience; and (3) conceptualize people as agentic and not simply as reactors to the environmental press. I illustrate those principles with research on domestic violence and welfare reform. In doing so, I call attention both to the way in which aspects of people's lives intersect with community settings, and to the embeddedness of people's lives (and community settings) in larger social structures. PMID- 11439830 TI - J. Wilbert Edgerton: introduction for the 2000 Recipient of the Society for Community Research and Action Award for Distinguished Contributions to Practice in Community Psychology. PMID- 11439831 TI - The community is it! PMID- 11439832 TI - Can home visitation enhance maternal social support? AB - This study investigates the impacts of Healthy Start, a statewide, home visitation program, on the social support systems of 212 disadvantaged mothers in Oahu, Hawaii. Maternal support was assessed at the time these mothers gave birth and 12 months later with the Maternal Social Support Index. Comparisons of mothers randomly assigned to visited (n = 108) or nonvisited (n = 104) conditions uncover few differences on structural or affiliational aspects of support after 12 months. Visited mothers, however, are significantly less likely to report a decline in satisfaction with a close adult than nonvisited mothers do. The discussion examines the challenges of fostering social support and the ability of home visitation efforts to achieve this outcome. PMID- 11439833 TI - The efficacy paradox in randomized controlled trials of CAM and elsewhere: beware of the placebo trap. PMID- 11439834 TI - Physicians, heal thyselves! PMID- 11439835 TI - Physicians, heal thyselves! PMID- 11439836 TI - Physicians heal thyselves! PMID- 11439837 TI - Physicians heal thyselves! PMID- 11439838 TI - Physicians heal thyselves! PMID- 11439839 TI - Physicians heal thyselves! PMID- 11439840 TI - Physicians heal thyselves! PMID- 11439841 TI - Physicians heal thyselves! PMID- 11439842 TI - Physicians heal thyselves! PMID- 11439843 TI - Treatments for young people with cancer. PMID- 11439844 TI - CordyMax Cs-4 improves steady-state bioenergy status in mouse liver. AB - OBJECTIVE: To evaluate effects of CordyMax Cs-4, a mycelial fermentation product of Cordyceps sinensis, on energy metabolism. DESIGN: An in vivo pharmacology study using 31P nuclear magnetic resonance (NMR) spectroscopy. SUBJECTS AND STUDY INTERVENTIONS: Adult male C57-BL/6 mice were given an aqueous extract of CordyMax, 200 or 400 mg/kg per day or placebo for 7 days. OUTCOME MEASUREMENTS: Using 31P-NMR spectroscopy to measure cellular triphosphates and inorganic phosphate, expressed as a ratio to a reference peak, and calculate tissue pH. RESULTS: Steady-state beta adenosine triphosphate (ATP) increased in the liver of mice that received CordyMax (200 or 400 mg/kg per day) for 7 days, by 12.3% +/- 0.8% and 18.4% +/- 0.9%, respectively, compared to placebo controls (both p < 0.001), suggesting a higher hepatic bioenergy status in CordyMax-treated animals. Hepatic inorganic phosphate (Pi) decreased by 24.5% +/- 0.9% and 17.6% +/- 1.7% in the two treatment groups, respectively, compared to placebo controls (p < 0.001). The ratio of beta-ATP:Pi increased by 47.7% +/- 1.6% and 41.4% +/- 2.4%, respectively, in the treatment groups (both p < 0.001 compared to placebo). After discontinuation of CordyMax for 7 days, beta-ATP and Pi returned towards baseline. CONCLUSION: CordyMax is effective in improving bioenergy status in the murine liver, suggesting a mechanism underlying the known clinical effectiveness of CordyMax in alleviating fatigue and improving physical endurance, especially in elderly subjects. PMID- 11439845 TI - Echinacea purpurea and melatonin augment natural-killer cells in leukemic mice and prolong life span. AB - OBJECTIVE: We recently showed that daily dietary administration of Echinacea purpurea root extract to normal mice for as little as 1 week resulted in significant elevations of natural-killer (NK) cells (immune cells that are cytolytic to virus-containing cells and many tumor cells). Such boosting of this fundamental immune cell population suggests a prophylactic role for this herb in normal animals. Based on this evidence, our goal in the present work was to assess the role of dietary administration of this herbal extract to mice bearing leukemia, a type of tumor well known to be a target for NK cells. DESIGN: A commercially available root extract of E. purpurea, which we have already shown to be highly effective in mice, was administered daily for 50 days from the onset of leukemia (day 0). Control leukemic mice received no extract. Other leukemic mice received the NK-enhancing neurohormone, melatonin, administered precisely as above. In all treatment and control categories, some mice were sampled at 9 days after tumor onset, others were sampled at 3 months, and still others were left to assess treatment effect on life span. RESULTS: At 9 days (intermediate stage leukemia; death beginning by day 17-18), E. purpurea-treated mice had a 2.5-fold increase in the absolute numbers of NK cells in their spleens. By 3 months after leukemia onset, E. purpurea-treated mice still had 2-3 times the normal numbers of NK cells in their spleens. No leukemic, untreated (control) mice remained alive at 3 months, hence the comparison with normal animals. Moreover, at 3 months post-tumor onset, all the major hemopoietic and immune cell lineages in their bone marrow birth site, were recorded at normal numbers, in E. purpurea consuming, leukemic mice. The survival advantage provided by administering these leukemic mice with E. purpurea was highly significant versus untreated, leukemic mice when analyzed by Kaplan-Meier survival statistics. CONCLUSION: The present study has provided the first systematic analysis, under controlled laboratory conditions, of the effect(s) of the botanical, E. purpurea, in vivo, in leukemic hosts. The profoundly positive effects of this herb in disease abatement observed in this study suggest the therapeutic potential of E. purpurea, at least with respect to leukemia, if not other tumors as well. PMID- 11439846 TI - Developing and evaluating complementary therapy services: Part 1. Establishing service provision through the use of evidence and consensus development. AB - The integration of complementary therapies within the British National Health Service (NHS) in the context of limited evidence of effectiveness has been much debated, as has the need for the provision of health services to be more evidence based. In June 1994, a project was launched within a South-East London NHS Hospital Trust to introduce complementary therapy (acupuncture, homeopathy, and osteopathy), in the context of an evaluation program. This followed approximately 4 years of working toward raising the profile of complementary therapies within the hospital through study days, workshops, and providing a massage and osteopathic service for staff. A survey of local general practitioners highlighted areas of complementary therapy provision and interest in referring patients to a hospital-based service. A steering group was established to draw together a proposal for funding the service. Evidence for the effectiveness of acupuncture, homeopathy, and osteopathy was presented at a multidisciplinary seminar. A consensus development process, using a modified Delphi technique to establish referral indicators followed this. This study provides a useful model of service development in the absence of good quality evidence for the effectiveness of clinical interventions. PMID- 11439847 TI - Patient perspectives on outcomes after treatment with acupuncture. AB - OBJECTIVES: To determine patients' experience of outcomes after acupuncture treatment. DESIGN: A patient questionnaire followed by semistructured interviews. SETTING: Acupuncture practices in York, England. SUBJECTS: Questionnaires were distributed by 4 acupuncturists to all 132 of their patients over a 2-week period, 72 (55%) of whom responded. Of these, 11 patients were interviewed. INTERVENTIONS: Patients received normal care from their acupuncturists, all of whom were members of the British Acupuncture Council. OUTCOME MEASURES: Patients reported on physical symptoms, emotional/mental symptoms, lifestyle changes, major life changes and inner life changes affecting outlook and attitudes to health. Patients also reported on the extent they attributed change to acupuncture and what they valued about attending for treatment. RESULTS: Primary reasons for attending were categorized by patients as physical symptoms (90%), mental and emotional problems (9%) and general health and well-being (1%). For physical symptoms, 75% of patients reported definite change; for emotional and mental symptoms, 67%; lifestyle changes, 40%, major life changes, 27%; and for inner life changes, 54%. Over time, many of the patients (42%) changed their primary reason for attending from their initial physical concerns, half of whom describing their new focus as related to general health and well-being. Some emotional changes were experienced by 83% of patients, irrespective of the initial reason for attending. For all these changes, 58% of patients reported that they "substantially" attributed the changes to acupuncture, and 25% "totally". The interviews provided additional data on the depth of the changes particularly for the longer term patients and on what they valued about treatment. CONCLUSIONS: Treatment with acupuncture results in a broad range of outcomes from physical and emotional change through to wider benefits involving the patients' lifestyle, outlook, and attitude toward their health. Existing outcome measures are not adequate to assess these wider benefits. The results suggest that patients experienced holistic benefits from acupuncture, an outcome that could be described as holism-in-action. PMID- 11439848 TI - A regional survey of health insurance coverage for complementary and alternative medicine: current status and future ramifications. AB - OBJECTIVE: The purpose of this survey is to evaluate the extent of health insurance coverage for complementary and alternative medicine (CAM) within one region in the United States, a study prompted by the increased utilization of CAM. DESIGN: Prospective telephone interview of health insurance representatives. LOCATION: A contiguous three-state area (New York, New Jersey, and Connecticut) in the North-east. RESULTS: Almost all of the insurers surveyed cover chiropractic services. Less than half of the insurers reimburse acupuncture, usually for chronic pain management. Coverage for massage therapy is minimal and usually associated with physical therapy or chiropractic treatment. Other CAM services receive negligible coverage. CONCLUSIONS: Current health insurance coverage of CAM is limited essentially to chiropractic medicine, acupuncture and massage therapy. Coverage of CAM is made confusing by different policies, practitioner requirements, and health plans within each carrier. PMID- 11439849 TI - Comment on Cleary-Guida et al.: CAM coverage. Survey outcomes pose more critical questions. PMID- 11439850 TI - Correlates of use of alternative medicine by the elderly in an urban population. AB - OBJECTIVE: To investigate the prevalence and predictors of use of complementary and alternative medicine (CAM) by the elderly. DESIGN: Cross-sectional survey examining patterns of use of complementary therapies in two urban multiethnic populations of older adults. SETTING AND SUBJECTS: A convenience sample of 421 older participants were interviewed at two sites: a university geriatrics primary care practice and a veterans medical clinic, both in New York City. Subjects were excluded if they did not speak English or if they were moderately cognitively impaired. MEASUREMENT: Use of CAM within the previous year. RESULTS: Fifty-eight percent (58%) of all subjects surveyed used some form of CAM, and close to 75% at the university practice alone. Use correlated most strongly with female gender (p < 0.0001), greater education (p = 0.0095), thyroid disease (p = 0.0190) and arthritis (p = 0.0251). There was no correlation with income, race, age, or self perceived health status. CONCLUSIONS: CAM use is highly prevalent in older persons in this study, especially among females and those who are more highly educated. PMID- 11439851 TI - Surveys of complementary and alternative medicine: Part II. Use of alternative and complementary cancer therapies. AB - The second part of this series on surveys of complementary and alternative medicine (CAM) in the United States provides a "point-of-information" summary of the studies on patients with cancer and their use of CAM therapies. Surveys of patients with cancer were the precursors of the recent wave of studies on CAM prevalence and use. Three tables summarize the findings from a total of 18 surveys categorized by Childhood Cancer, Adult Cancer, and Breast Cancer studies. PMID- 11439852 TI - Symposium showcases continued development of acupuncture research. PMID- 11439853 TI - National Population Policy 2000: an insight. PMID- 11439854 TI - Polio eradication--a success in sight. AB - India took a giant step closer to eradicating polio through the strategy of National Immunization Days-PPI. In order to validate the reported coverage for 1999-2000, UNICEF had undertaken the CES for PPI on the request from GOI. The paper is a presentation of findings from 15 states, carrying the data as of Aug. 31, 2000. The modified cluster sampling has been used in this study. Data reveals that out of 15, 10 states have more than 95% coverage for at least 3 doses, MP and Gujarat at the top with 99% coverage. Despite this high coverage level, huge number of children is still unreached. Nearly 5 lakh children are left out in UP alone. Ironically, higher proportion of urban clusters have zero dose children. Apart from lack of awareness about date and need of additional doses, lack of positive attitude among parents are major cause of not getting any of the doses. Qualitative research pinpointed some more reasons for non-compliance- apprehension about side effect, knowledge and traditional barriers. According to the service providers and influencers, lack of proper training and monetary compensation are major demotivating factors. PMID- 11439855 TI - Missed opportunities for immunisation in children. AB - All opportunities for immunisation in children should be utilised properly, as sustaining high levels of full immunisation coverage is essential to meet the goal of eradication of poliomyelitis as well as control the other vaccine preventable diseases; yet many opportunities for immunisation are missed in all types of health facilities, even in teaching hospitals. Reducing such missed opportunities is the cheapest way to increase immunisation coverage. The present study discusses the problem of missed opportunities for immunisation in children in Paediatric Outpatient Department and Immunisation Clinic of R.G. Kar Medical and Hospital, Calcutta and the underlying factors of the problem. Prevalence of missed opportunities in Paediatric OPD and Immunisation clinic was 37.8% and 1.4% respectively. Most of the missed opportunities were attributed to health care providers and delivery system of health care of the studied hospital. PMID- 11439856 TI - Maternal age as a risk factor for stillbirth. AB - To Detect the risk of still birth in different maternal age, five year (1st January 1993 to 31st December 1997) record of Government Medical College, Nagpur were analysed. The rate of stillbirth was 2.5% (1138 stillbirth out of 46,443 deliveries) which was significantly associated with increasing maternal age (x2 = 182.3, df = 4, p < 0.001). Teenage pregnancy and elderly pregnancy were in significant risk as compared to 20-29 years age group for stillbirth (OR = 1.6, 95% C1 1.1-2.5; OR 2.6, 95% CI = 1.9-3.5 respectively). PMID- 11439857 TI - A study on certain correlates in respect to the sources of awareness about pulse polio immunisation (1996-97). PMID- 11439858 TI - Canine balantidiasis--its treatment, epidemiological and zoonotic significance. PMID- 11439859 TI - Health appraisal of 1-6 years children in slum population of Ghaziabad City. PMID- 11439860 TI - Strategies for eradication of poliomyelitis--the Indian experience. AB - Substantial progress has been made towards achieving global eradication of poliomyelitis by the end of the year 2000; the goal set by the World Health Assembly in May 1988. The basic strategies to eradicate polio are: attaining high routine coverage with at least three doses of OPV; conducting national immunization days (NIDs) in polio endemic countries; establishing a sensitive system of acute flaccid paralysis (AFP) surveillance to track wild poliovirus circulation; and conducting "mopping-up" immunization when polio is reduced to focal transmission. By the end of 2000. India was in the midst of the sixth National Immunization Days (NIDs). Surveillance system for Acute Flaccid Paralysis (AFP) continued to achieve the recommended non-polio AFP rate of at least 1 per 100,000 population < 15 years per year (1.88 for week 51 ending 23rd December 2000), the adequate stool specimen collection rate was 83% that met the target of > 80%. Some States in the south and west have started to implement mopping-up immunization campaigns as the end-stage strategy to eliminate the last remaining foci of transmission. While most of India appears to be well placed to eradicate polio by the end of 2000 or shortly thereafter, concerns remain about low coverage in parts of the densely populated northern States of Uttar Pradesh and Bihar where high intensity transmission persists. The Government of India has embarked upon an intensified strategy that relies on extra rounds of NIDs; house to-house immunization to reach previously missed children; and aggressive mopping up campaigns including pre-emptive mopping-up in the known reservoirs in Uttar Pradesh and Bihar. Extensive microplanning and supervision of the supplementary immunization activities is critical to achieve the target of polio eradication. PMID- 11439861 TI - Initiatives for promotion of breast feeding. PMID- 11439862 TI - Zoonoses: challenges and strategies. AB - Zoonoses account for about four fifth of the human infections. The health problems posed by these seem to be increasing. Newer problems are recognised. Older and endemic problems appear in different forms. Zoonosis management is getting complex and complicated. Attention on their control and eradication is essentially required. PMID- 11439863 TI - Qualitative research: a tool in community based health care studies: some issues. PMID- 11439864 TI - A rapid assessment of anaemia in pregnancy in West Bengal with special reference to care seeking behaviour of mothers. AB - A cross-sectional study was conducted among the pregnant women in three administrative divisions of West Bengal to undertake a rapid assessment about the magnitude of the problem of anaemia in pregnancy and also to study care seeking behaviour for the same. The findings revealed that the occurrence of anaemia in these three divisions were very high to the extent of 86.39% and popularly known as 'Raktasunyata" or 'Raktalpata' to the common people. As per WHO guidelines (< 40% prevalence) it could be considered as public health problem of very high magnitude. Caregivers prescribed iron supplementation only in 70% of registered pregnant women when 100% coverage of pregnant women with IFA tablet is our national goal. Amongst this group 72.2% were partially consuming these tablets. Main reason for irregular or partial consumption was inability to purchase iron tablets (52.63%). Around 16% mothers were taking rest for 2 hours at daytime during pregnancy. Most important reasons for not consuming iron tablets were that iron was prescribed on the day of study (43.18%) and iron was not prescribed even though mothers were registered (36.23%). Focus group discussion highlighted some factors regarding improvement of the situation. PMID- 11439865 TI - A study on breastfeeding practices in a rural coal mine area of Andhra Pradesh. PMID- 11439866 TI - An evaluation of re-orientation training program of ICDS functionaries on protein energy malnutrition. PMID- 11439867 TI - ORBIS International in India. PMID- 11439868 TI - Chrysotile asbestos: why should a carcinogen remain in demand? PMID- 11439869 TI - Presidential address. PMID- 11439870 TI - Dr. K. N. Rao Memorial Oration. Public health--core values in the changing world. PMID- 11439871 TI - Rapid assessment of cataract blindness in India. AB - 28,055 persons aged 50 yrs+ from seven states in India were surveyed by a rapid assessment technique for cataract blindness. The prevalence of bilateral blindness (vision < 6/60 in the better eye) was 11.68 percent (95% C.I. 10.54 12.81). The age-gender adjusted blindness prevalence rate was 11.04 percent (95% C.I. 11.033-11.044). Age and occupational status were associated with blindness prevalence. Cataract was the commonest cause of low vision and blindness in this population. Respondents aged 60-69 years had a 2.74 times higher risk, while those aged 70 years+ had a 4.86 times higher risk of being blind, compared to those 50-59 years. Productively employed individuals had lowest blindness rates. Blindness rates were five times higher among respondents who were not working and two times higher among those engaged solely in household activities. The prevalence of cataract was 43.32 percent (95% C.I. 41.14-45.50) among those aged 50+ years. The prevalence increased with increasing age. Gender did not influence the prevalence of cataract in the present survey. Extrapolating from the present survey, it is estimated that 11.9 million blind people (vision < 6/60 in the better eye) in India are in urgent need of cataract surgery. PMID- 11439872 TI - A survey on refractive error and strabismus among children in a school at Aligarh. AB - An ophthalmic survey of 310 school going children in the age group of 4-12 years was done. 41 children (13.2%) were found to have a vision of less than 6/6 in one or both eyes. Myopia of 0.5 D or more and hyperopia of +2 D or more or astigmatism of 1 D or more in one or both eyes were seen in 74 children (22.9%). There was a tendency for a decrease in hyperopia and an increase in myopia with advancing age. Astigmatic error of 1 D or less was seen in 12 eyes (1.93%) and anisometropia of more than 1 D was present in 11 cases (3.5%). Hypertropia was more in female children whereas myopia had a nearly equal distribution in both sexes. There was heterophoria and manifest strabismus which showed a gradual increase with advancing age. This study was compared to few other studies undertaken in the past. PMID- 11439873 TI - Disaster to development. PMID- 11439874 TI - Extents of contamination of top milk and their determinants in an urban slum of Varanasi, India. AB - A community based study to examine the extent of contamination of supplementary milk feeds of 149 children aged 6-24 months was conducted in a semi urban slum of Varanasi, India. Out of 201 children, 149 top milk samples were collected directly from the feeding utensils into a sterile vial and subjected to bacteriological analysis. Overall, 53.7% of milk samples were contaminated by bacteria and among them 16.1% were potentially enteropathogenic in nature. The distribution of pathogens was E. coli (13.4%), Klebsiella spp (5.4%), Enterobacter spp. (5.4%), Pseudomonas aeruginosa (4.7%), Shigella spp. (2.7%) and others (22.1%). The rate of contamination was significantly higher (p < 0.001) in lower income group (73.4%), lower caste (69.6%) and in case of illiterate mothers (69.3%). Bivariate analysis indicated that wherever the afore mentioned parameters of hygiene were adverse, isolation rates increased multifoldely. Multiple logistic regression analysis indicated that the probability of a milk sample being positive for bacterial contamination was higher by 20 times when unclean utensils were used, by 3 times if mothers hands were dirty and by 2.8 time if the mothers were illiterate. The odds of contamination by pathogens was 25.7 times higher if the feeding utensils were dirty. PMID- 11439875 TI - Effect of continuing training on knowledge and practices of traditional birth attendants about maternal and newborn care. AB - Seventy nine traditional birth attendants (TBAs) of Raipur Rani community development block, Haryana were interviewed to assess the effectiveness of continuing training in changing their knowledge and practices regarding maternal and newborn care. Seventy three percent of them reported participation in continuing training sessions. However, analysis of attendance register showed that only 35.4% had attended more than 50% sessions in year 1993. Most (83.5%) of the TBAs gave advice to pregnant women for increased food intake, 47% advised tetanus toxoid, 16.5% for more rest, and 31.6% for iron tablets. Many of them were aware of maternal complications i.e. anaemia (64.6%), oedema (26.6%), bleeding per veginum (39.2%), abnormal presentation (77.2%) and high fever (48.1%). Risks to newborn like low birth weight, fever, cough/rapid breathing and hypothermia were known to 20.2%, 31.6%, 17.7% and 1.3% of the TBAs respectively. Knowledge regarding causes of low birth weight baby like 'weak' mother, less diet in pregnancy, short birth interval and preterm delivery were reported by 69.6%, 63.3%, 12.6% and 3.8% respectively. About two fifth of TBAs advised referral to hospital in case of prolonged labour and 88.6% for very low birth weight babies. Disposable Dai Kit and weighing machine were available with 32% and 73% TBAs. Significantly higher proportion of TBAs participating in continuing training advised tetanus toxoid vaccination, appropriate feeding practices of the newborn, hospital referral in case of prolonged labour and were less inclined to advise injection to speed up labour. Therefore, efforts should be made to increase the attendance of TBAs in continuing training sessions so as to sustain modern maternal and newborn care practices acquired after initial training. PMID- 11439876 TI - Diet nutrition and health profile of elderly population of urban Baroda. AB - Geriatric population forms a significant proportion of our total population. Hence, various problems affecting the overall health of the elderly need special consideration. In this context, studies were undertaken to assess the socio demographic factors, diet and health profile of 320 elderly men and women of all the three income groups of Urban Baroda. Data on socio-demographic factors was collected using an open ended questionnaire. Nutritional status was assessed using anthropometric measurements of height, weight, mid upper arm circumference (MUAC) and body mass index (BMI). Information on dietary profile was collected by 24 hour dietary recall method. Fasting practices were also studied. Socio demographic data of geriatric men of high, middle and low income groups revealed that majority of the subjects were married. A greater percentage of high income group (HIG) men had nuclear family whereas majority of low income groups (LIG) elderly men resided in a joint family. Socio-demographic profile of elderly women of all the 3 income groups revealed that most of the subjects were Hindus. The percentage of widowhood, illiteracy and joint family system were higher in LIG as compared to the elderly women in middle and high income groups (MIG and HIG). Nutrient intake data of elderly men of all the income groups revealed lower consumption of energy, protein, iron and beta-carotene as compared to the RDA whereas fats and vitamin C intakes were higher as compared to the RDA (p < 0.05). The mean nutrient intake, by the LIG elderly women, in terms of energy, protein, iron, calcium, beta-carotene and vitamin C were significantly (p < 0.05) lower than the RDA as well as when compared to the elderly women of MIG and HIG. Mean anthropometric measurements of weight and BMI were higher in elderly HIG and MIG men as compared to the elderly men from LIG. Significant difference was found in all the anthropometric measurements of elderly women of LIG as compared to HIG and MIG. Morbidity profile showed a striking rise in problems of oral cavity, cardio vascular disease, neurological problems and problems of gastro intestinal tract with advancing age in both elderly men and women. The study reveals striking differences in diet, health and disease profile with advancing age. PMID- 11439877 TI - Increasing positivity of HIV antibodies among the blood donors of Howrah district. PMID- 11439878 TI - Knowledge, attitudes and practices (KAP) regarding breast feeding at Chandigarh. PMID- 11439879 TI - A study on community awareness on child immunisation. PMID- 11439880 TI - Breast feeding practices in rural areas of West Bengal. PMID- 11439881 TI - After two years, did Viagra live up to its expectations? AB - Sildenafil (Viagra) is the first on-demand oral medication approved for treatment of male erectile dysfunction (ED). Since the unprecedented release of sildenafil, the initial surge of 'Viagra craze' has subsided and there is a considerable decline in world-wide prescriptions. We did a long-term efficacy study of sildenafil in our community practice to assess the discontinuation rate and the etiologic factors causing discontinuation. PMID- 11439882 TI - Shortcuts: the savings may be costly. PMID- 11439883 TI - [Therapeutic effects of macrolides on patients with psoriasis vulgaris, pigmentary prurigo, or eosinophilic pustule folliculitis]. PMID- 11439884 TI - [Immunomodulating effects of roxithromycin on skin]. PMID- 11439885 TI - [Therapeutic effects of macrolides on patients with palmoplantar pustulosis]. PMID- 11439886 TI - [Effects of macrolide therapy on patients with acne vulgaris]. PMID- 11439887 TI - [Case of the bare lymphocyte syndrome successfully treated with long-term small dosage administration of erythromycin]. PMID- 11439888 TI - [Etiological mechanism of long QT syndrome and the effects of macrolides]. PMID- 11439889 TI - [Calcium dependent effect of clarithromycin on IL-8 production in cultured human epidermal cells]. PMID- 11439890 TI - [Effects of erythromycin on the Pseudomonas-induced rabbit diffuse panbronchiolitis model--the improvements in granuloma-like lesions with increasing production of MCP-1]. PMID- 11439891 TI - [Effects of macrolides on cultured rat vascular cells]. PMID- 11439892 TI - [Effects of roxithromycin on experimental otitis media]. PMID- 11439893 TI - [Effects of erythromycin on matrix metalloproteinase-9(MMP-9)and cell migration]. PMID- 11439894 TI - [Effects of macrolide antibiotics on immune cells derived from human peripheral blood]. PMID- 11439895 TI - [Effect of macrolide antibiotics on apoptosis induction]. PMID- 11439896 TI - [Effect of roxithromycin on induction of apoptosis in peripheral blood lymphocytes from patients with bronchial asthma]. PMID- 11439897 TI - [Effects of macrolides on neutrophil apoptosis, IL-8 production, and neutrophil clearance mechanism]. PMID- 11439898 TI - [Enhancement of glucocorticoid production by roxithromycin]. PMID- 11439899 TI - [Comparative study of effects of anti-infective and respiratory system agents on endogeneous glucocorticoids with macrolides]. PMID- 11439900 TI - [Guidelines for small dosage macrolide therapy of patients with diffuse panbronchiolitis and reports of adverse effects]. PMID- 11439901 TI - [Effect and action mechanism of short-term administration of clarithromycin for airway hypersecretion]. PMID- 11439902 TI - [Effect of macrolides on rat models of airway hypersecretion]. PMID- 11439903 TI - [Suppression of non-small-cell lung carcinoma metastasis with clarithromycin]. PMID- 11439904 TI - [Suppression of biological response to surgical stress of lung cancer resection by administration of 14-cyclol linkage macrolides]. PMID- 11439905 TI - [P-glycoprotein inhibition by macrolide antibiotics: The effects on resistance of cancer cells to antineoplastic agents and pharmacokinetics of the agents as P glycoprotein substrate]. PMID- 11439906 TI - [Suppressive efffects of clarithromycin and cyclooxygenase-2 inhibitor, etodolac on N-nitrosobis(2-hydroxypropyl)amine(BHP)- induced experimental lung neoplasm in rats]. PMID- 11439907 TI - [Effects and action mechanisms of macrolides on IgG- immune complex lung injury models]. PMID- 11439908 TI - [Suppressive effect of 14-cyclol linkage macrolides on bleomycin-induced pulmonary fibrosis]. PMID- 11439909 TI - [Effect of rapamycin on cytoplasmic accumulation of hetero nuclear ribonucleoprotein C-like unwinding protein 2]. PMID- 11439910 TI - [Effects of various erythromycin derivatives on activation of transcription factors in airway epithelial cells]. PMID- 11439911 TI - Pyruvate-based dialysate preserves neutrophilic oxygen consumption. PMID- 11439912 TI - American Academy of Pediatric Dentistry. Reference manual 2000-01. PMID- 11439913 TI - A late developing mandibular premolar supernumerary tooth. AB - This paper presents a case of a patient who developed a mandibular premolar supernumerary tooth between the age of 11 and 20 years. Evidence for the late development of the supernumerary tooth comes from consecutive panoramic radiographs. A review of the international literature concerning late developing supernumerary teeth is included. PMID- 11439914 TI - 24th annual meeting of the International Urogynecological Association (IUGA). A joint meeting with the International Continence Society (ICS) and the International Children's Continence Society (ICCS). August 23-26, 1999, Denver, Colorado, USA. Abstracts. PMID- 11439915 TI - LDA President: Dr. Michael J. Haight. Interview by Leigh T. Berry. PMID- 11439916 TI - Selling or closing a dental practice: some risk management considerations. PMID- 11439917 TI - Saving the life of a practice after the death of the dentist. PMID- 11439918 TI - A new pedigree with recessive mapping to CHED2 locus on 20p13. PMID- 11439919 TI - 2nd International Symposium on the Molecular Biology of Breast Cancer. Lillehammer, Norway, 12-16 March 2000. Abstracts. PMID- 11439920 TI - 35th annual meeting of the European Society for Clinical Investigation. Barcelona, Spain, 2-5 May 2001. Abstracts. PMID- 11439921 TI - Bestor: CFOs with expanded skills are good CEO candidates. PMID- 11439922 TI - Implications of the new privacy standards for healthcare institutions. PMID- 11439923 TI - Doing the Texas "to-do" steps. PMID- 11439924 TI - [Patient records and sale of practice]. PMID- 11439925 TI - [Legal pitfalls in clinical practice and their solutions. The physician between laws of economics and standards of care]. PMID- 11439926 TI - Ziprasidone approved. PMID- 11439927 TI - Reader mail. Glycemic load. PMID- 11439928 TI - ATP-dependent activation of the intermediate conductance, Ca2+-activated K+ channel, hIK1, is conferred by a C-terminal domain. AB - We previously demonstrated that hIK1 is activated directly by ATP in excised, inside-out patches in a protein kinase A inhibitor 5-24 dependent manner, suggesting a role for phosphorylation in the regulation of this Ca(2+)-dependent channel. However, mutation of the single consensus cAMP-dependent protein kinase phosphorylation site (S334A) failed to modify the response of hIK1 to ATP (Gerlach, A. C., Gangopadhyay, N. N., and Devor, D. C. (2000) J. Biol. Chem. 275, 585-598). Here we demonstrate that ATP does not similarly activate the highly homologous Ca(2+)-dependent K(+) channels, hSK1, rSK2, and rSK3. To define the region of hIK1 responsible for the ATP-dependent regulation, we generated a series of hIK1 truncations and hIK1/rSK2 chimeras. ATP did not activate a chimera containing the N terminus plus S1-S4 from hIK1. In contrast, ATP activated a chimera containing the hIK1 C-terminal amino acids His(299)-Lys(427). Furthermore, truncation of hIK1 at Leu(414) resulted in an ATP-dependent channel, whereas larger truncations of hIK1 failed to express. Additional hIK1/rSK2 chimeras defined the minimal region of hIK1 required to confer complete ATP sensitivity as including amino acids Arg(355)-Ala(413). An alanine scan of all non-conserved serines and threonines within this region failed to alter the response of hIK1 to ATP, suggesting that hIK1 itself is not directly phosphorylated. Additionally, substitution of amino acids Arg(355)-Met(368) of hIK1 into the corresponding region of rSK2 resulted in an ATP-dependent activation, which was approximately 50% of that of hIK1. These results demonstrate that amino acids Arg(355)-Ala(413) within the C terminus of hIK1 confer sensitivity to ATP. Finally, we demonstrate that the ATP-dependent phosphorylation of hIK1 or an associated protein is independent of Ca(2+). PMID- 11439929 TI - Biosynthetic controls on the 13C contents of organic components in the photoautotrophic bacterium Chloroflexus aurantiacus. AB - To assess the effects related to known and proposed biosynthetic pathways on the (13)C content of lipids and storage products of the photoautotrophic bacterium Chloroflexus aurantiacus, the isotopic compositions of bulk cell material, alkyl and isoprenoid lipids, and storage products such as glycogen and polyhydroxyalkanoic acids have been investigated. The bulk cell material was 13 per thousand depleted in (13)C relative to the dissolved inorganic carbon. Evidently, inorganic carbon fixation by the main carboxylating enzymes used by C. aurantiacus, which are assumed to use bicarbonate rather than CO(2), results in a relatively small carbon isotopic fractionation compared with CO(2) fixation by the Calvin cycle. Even carbon numbered fatty acids, odd carbon numbered fatty acids, and isoprenoid lipids were 14, 15, and 17-18 per thousand depleted in (13)C relative to the carbon source, respectively. Based on the (13)C contents of alkyl and isoprenoid lipids, a 40 per thousand difference in (13)C content between the carboxyl and methyl carbon from acetyl-coenzyme A has been calculated. Both sugars and polyhydroxyalkanoic acid were enriched in (13)C relative to the alkyl and isoprenoid lipids. To the best of our knowledge this is the first report in which the stable carbon isotopic composition of a large range of biosynthetic products in a photoautotrophic organism has been investigated and interpreted based on previously proposed inorganic carbon fixation and biosynthetic pathways. Our results indicate that compound-specific stable carbon isotope analysis may provide a rapid screening tool for carbon fixation pathways. PMID- 11439931 TI - Stress-strain relations in embryonic chick heart. PMID- 11439930 TI - Conversion of the ion selectivity of the 5-HT(3a) receptor from cationic to anionic reveals a conserved feature of the ligand-gated ion channel superfamily. AB - The 5-hydroxytryptamine(3) (5-HT(3)) receptor is a member of a superfamily of ligand-gated ion channels, which includes nicotinic acetylcholine, gamma aminobutyric acid, and glycine receptors. The receptors are either cation or anion selective, leading to their distinctive involvement in either excitatory or inhibitory neurotransmission. Using a combination of site-directed mutagenesis and electrophysiological characterization of homomeric 5-HT(3A) receptors expressed in HEK293 cells, we have identified a set of mutations that convert the ion selectivity of the 5-HT(3A) receptor from cationic to anionic; these were substitution of V13'T in M2 together with neutralization of glutamate residues (E 1'A) and the adjacent insertion of a proline residue (P-1') in the M1-M2 loop. Mutant receptors showed significant chloride permeability (P(Cl)/P(Na) = 12.3, P(Na)/P(Cl) = 0.08), whereas WT receptors are predominantly permeable to sodium (P(Na)/P(Cl) > 20, P(Cl)/P(Na) < 0.05). Since the equivalent mutations have previously been shown to convert alpha7 nicotinic acetylcholine receptors from cationic to anionic (Galzi J.-L., Devillers-Thiery, A, Hussy, N., Bertrand, S. Changeux, J. P., and Bertrand, D. (1992) Nature 359, 500-505) and, recently, the converse mutations have allowed the construction of a cation selective glycine receptor (Keramidas, A., Moorhouse, A. J., French, C. R., Schofield, P. R., and Barry, P. H. (2000) Biophys. J. 78, 247-259), it appears that the determinants of ion selectivity represent a conserved feature of the ligand-gated ion channel superfamily. PMID- 11439932 TI - Web alert. PMID- 11439934 TI - Scholarship and the RO1. PMID- 11439935 TI - Appropriate use of pilot studies. PMID- 11439937 TI - 4th International Conference on Bipolar Disorder. June 14-16, 2001, Pittsburgh, Pennsylvania. Abstracts. PMID- 11439936 TI - Central neuronal fatigue in multiple sclerosis. PMID- 11439938 TI - From the Centers for Disease Control and Prevention. Protracted outbreaks of cryptosporidiosis associated with swimming pool use--Ohio and Nebraska, 2000. PMID- 11439939 TI - From the Centers for Disease Control and Prevention. Prevalence of parasites in fecal material from chlorinated swimming pools--United States, 1999. PMID- 11439940 TI - JAMA patient page. Spina bifida. PMID- 11439941 TI - 5th International Workshop on HIV Drug Resistance & Treatment Strategies. 4-8 June 2001, Scottsdale, Arizona. Abstracts. PMID- 11439942 TI - 15th International Conference on Oral & Maxillofacial Surgery. Durban, South Africa, 19-24 May 2001. Abstracts. PMID- 11439943 TI - The clinical spectrum of familial hemiplegic migraine associated with mutations in a neuronal calcium channel. AB - BACKGROUND: Familial hemiplegic migraine, an autosomal dominant disorder characterized by attacks of transient hemiparesis followed by a migraine headache, is classically divided into pure familial hemiplegic migraine (affecting 80 percent of families) and familial hemiplegic migraine with permanent cerebellar signs (affecting 20 percent of families). Mutations in CACNA1A, which encodes a neuronal calcium channel, are present in 50 percent of families with hemiplegic migraine, including all those with cerebellar signs. We studied the various clinical manifestations associated with mutations in CACNA1A in families with hemiplegic migraine with and without cerebellar signs. METHODS: CACNA1A was analyzed and nine mutations were detected in 15 of 16 probands of families affected by hemiplegic migraine and cerebellar signs, in 2 of 3 subjects with sporadic hemiplegic migraine and cerebellar signs, and in 4 of 12 probands of families affected by pure hemiplegic migraine. Genotyping of probands and relatives identified a total of 117 subjects with mutations whose clinical manifestations were assessed in detail. RESULTS: Eighty-nine percent of the subjects with mutations had attacks of hemiplegic migraine. One third had severe attacks with coma, prolonged hemiplegia, or both, with full recovery. All nine mutations, including five newly identified ones, were missense mutations. Six mutations were associated with hemiplegic migraine and cerebellar signs, and 83 percent of the subjects with these six mutations had nystagmus, ataxia, or both. Three mutations were associated with pure hemiplegic migraine. CONCLUSIONS: Hemiplegic migraine in subjects with mutations in CACNA1A has a broad clinical spectrum. This clinical variability is partially associated with the various types of mutations. PMID- 11439944 TI - Improvement in cardiac function in the cardiac variant of Fabry's disease with galactose-infusion therapy. PMID- 11439945 TI - Risk of uterine rupture during labor among women with a prior cesarean delivery. AB - BACKGROUND: Each year in the United States, approximately 60 percent of women with a prior cesarean delivery who become pregnant again attempt labor. Concern persists that a trial of labor may increase the risk of uterine rupture, an uncommon but serious obstetrical complication. METHODS: We conducted a population based, retrospective cohort analysis using data from all primiparous women who gave birth to live singleton infants by cesarean section in civilian hospitals in Washington State from 1987 through 1996 and who delivered a second singleton child during the same period (a total of 20,095 women). We assessed the risk of uterine rupture for deliveries with spontaneous onset of labor, those with labor induced by prostaglandins, and those in which labor was induced by other means; these three groups of deliveries were compared with repeated cesarean delivery without labor. RESULTS: Uterine rupture occurred at a rate of 1.6 per 1000 among women with repeated cesarean delivery without labor (11 women), 5.2 per 1000 among women with spontaneous onset of labor (56 women), 7.7 per 1000 among women whose labor was induced without prostaglandins (15 women), and 24.5 per 1000 among women with prostaglandin-induced labor (9 women). As compared with the risk in women with repeated cesarean delivery without labor, uterine rupture was more likely among women with spontaneous onset of labor (relative risk, 3.3; 95 percent confidence interval, 1.8 to 6.0), induction of labor without prostaglandins (relative risk, 4.9; 95 percent confidence interval, 2.4 to 9.7), and induction with prostaglandins (relative risk, 15.6; 95 percent confidence interval, 8.1 to 30.0). CONCLUSIONS: For women with one prior cesarean delivery, the risk of uterine rupture is higher among those whose labor is induced than among those with repeated cesarean delivery without labor. Labor induced with a prostaglandin confers the highest risk. PMID- 11439946 TI - Images in clinical medicine. Vomiting of neurologic origin. PMID- 11439947 TI - Clinical practice. Postmenopausal hormone-replacement therapy. PMID- 11439948 TI - Hepatitis C virus infection. PMID- 11439949 TI - Vaginal delivery after cesarean section--is the risk acceptable? PMID- 11439950 TI - New therapies for Fabry's disease. PMID- 11439951 TI - Hemiplegic migraine--downstream of a single-base change. PMID- 11439952 TI - The death of young King Edward VI. PMID- 11439953 TI - Retraction: reconstitution of hematopoiesis after high-dose chemotherapy by autologous progenitor cells generated ex vivo. PMID- 11439954 TI - Malabsorption due to cholecystokinin deficiency in a patient with autoimmune polyglandular syndrome type I. PMID- 11439955 TI - Malabsorption due to cholecystokinin deficiency in a patient with autoimmune polyglandular syndrome type I. PMID- 11439956 TI - Lack of effect of induction of hypothermia after acute brain injury. PMID- 11439957 TI - Lack of effect of induction of hypothermia after acute brain injury. PMID- 11439958 TI - Vitamin D intoxication associated with an over-the-counter supplement. PMID- 11439959 TI - Preventing upper gastrointestinal bleeding in patients with Helicobacter pylori infection. PMID- 11439960 TI - The cost effectiveness of antiretroviral therapy for HIV disease. PMID- 11439961 TI - The cost effectiveness of antiretroviral therapy for HIV disease. PMID- 11439962 TI - The disposition of unused frozen embryos. PMID- 11439963 TI - Safety and efficacy of recombinant human alpha-galactosidase A replacement therapy in Fabry's disease. AB - BACKGROUND: Fabry's disease, lysosomal alpha-galactosidase A deficiency, results from the progressive accumulation of globotriaosylceramide and related glycosphingolipids. Affected patients have microvascular disease of the kidneys, heart, and brain. METHODS: We evaluated the safety and effectiveness of recombinant alpha-galactosidase A in a multicenter, randomized, placebo controlled, double-blind study of 58 patients who were treated every 2 weeks for 20 weeks. Thereafter, all patients received recombinant alpha-galactosidase A in an open-label extension study. The primary efficacy end point was the percentage of patients in whom renal microvascular endothelial deposits of globotriaosylceramide were cleared (reduced to normal or near-normal levels). We also evaluated the histologic clearance of microvascular endothelial deposits of globotriaosylceramide in the endomyocardium and skin, as well as changes in the level of pain and the quality of life. RESULTS: In the double-blind study, 20 of the 29 patients in the recombinant alpha-galactosidase A group (69 percent) had no microvascular endothelial deposits of globotriaosylceramide after 20 weeks, as compared with none of the 29 patients in the placebo group (P<0.001). Patients in the recombinant alpha-galactosidase A group also had decreased microvascular endothelial deposits of globotriaosylceramide in the skin (P<0.001) and heart (P<0.001). Plasma levels of globotriaosylceramide were directly correlated with clearance of the microvascular deposits. After six months of open-label therapy, all patients in the former placebo group and 98 percent of patients in the former recombinant alpha-galactosidase A group who had biopsies had clearance of microvascular endothelial deposits of globotriaosylceramide. The incidence of most treatment-related adverse events was similar in the two groups, with the exception of mild-to-moderate infusion reactions (i.e., rigors and fever), which were more common in the recombinant alpha-galactosidase A group. IgG seroconversion occurred in 88 percent of patients who received recombinant alpha galactosidase A. CONCLUSIONS: Recombinant alpha-galactosidase A replacement therapy cleared microvascular endothelial deposits of globotriaosylceramide from the kidneys, heart, and skin in patients with Fabry's disease, reversing the pathogenesis of the chief clinical manifestations of this disease. PMID- 11439964 TI - Accessory middle cerebral artery and moyamoya disease. PMID- 11439965 TI - Maternal age is not a risk factor for Parkinson's disease. PMID- 11439966 TI - Hippocampal involvement in identical twins with neurofibromatosis type 1. PMID- 11439967 TI - Paroxysmal hypertension during a complex partial seizure. PMID- 11439968 TI - Compression by looping and perforation of the facial nerve by the anterior inferior cerebellar artery: a possible cause of facial tic. PMID- 11439969 TI - Charles Bonnet syndrome; an example of cortical dissociation syndrome affecting vision? PMID- 11439970 TI - Effects of topiramate on cognition. PMID- 11439971 TI - Postoperative hearing loss due to venous congestion at the inferior colliculus, or cochlear dysfunction? PMID- 11439972 TI - 6th Latin American Congress on Sexual Impotence and 6th Brazilian Congress on Sexual Impotence. 13-16 June 2001, Rio de Janeiro, Brazil. Abstracts. PMID- 11439973 TI - 7th International Symposium on New Quinolones. Edinburgh, UK, 10-12 June 2001. Abstracts. PMID- 11439974 TI - T-cell clonality of undetermined significance. PMID- 11439975 TI - Homozygous gene conversion in von Willebrand factor gene as a cause of type 3 von Willebrand disease and predisposition to inhibitor development. PMID- 11439976 TI - A rapid and reliable 7-deletion multiplex polymerase chain reaction assay for alpha-thalassemia. PMID- 11439977 TI - Improve or abandon the standardized response criteria for myelodysplastic syndromes recommended by the International Working Group. PMID- 11439979 TI - Dutch ship fails to offer Irish women abortions. PMID- 11439980 TI - Abortion in Northern Ireland to be reviewed. PMID- 11439981 TI - Report suggests small link between depleted uranium and cancer. PMID- 11439982 TI - UK body supports chemotherapy drugs for lung cancer. PMID- 11439983 TI - Israeli psychiatrists propose new law on hospital admissions. PMID- 11439984 TI - Cancer survival rates continue to rise in the Netherlands. PMID- 11439985 TI - Glaxo cuts drug prices for developing countries. PMID- 11439986 TI - Medicine information leaflets fail concordance test. PMID- 11439987 TI - What have we learnt from the Alder Hey affair? February 2001 seems to have been average month for organ donations in Newcastle. PMID- 11439988 TI - What have we learnt from the Alder Hey affair? Lack of information on transplant procedures is disturbing. PMID- 11439989 TI - What have we learnt from the Alder Hey affair? Criticism of pathologists has been unfair. PMID- 11439990 TI - What have we learnt from the Alder Hey affair? Clinicians and pathologists must work as team. PMID- 11439991 TI - What have we learnt from the Alder Hey affair? Asking for consent would halt decline in voluntary necropsies. PMID- 11439992 TI - alpha Streptococci and recurrences of otitis media. Right choice of antibiotic can decrease risk of recurrence. PMID- 11439993 TI - alpha Streptococci and recurrences of otitis media. Tampering with microbial ecology is risky. PMID- 11439994 TI - Patients' preferences for patient centred approach to consultation. What is patient centredness? PMID- 11439995 TI - BSE and variant CJD. Assumption that BSE originated from scrapie in sheep led to misjudgment. PMID- 11439996 TI - BSE and variant CJD. Humans can live with BSE so long as they do not eat brains. PMID- 11439997 TI - Patient education to encourage graded exercise in chronic fatigue syndrome. Trial has too many shortcomings. PMID- 11439998 TI - Higher dose inhaled steroids in childhood asthma. Conventional doses do have side effects. PMID- 11439999 TI - Higher dose inhaled steroids in childhood asthma. Why isn't titration advocated more often in delivery of inhaled drugs? PMID- 11440000 TI - Thromboprophylaxis after replacement arthroplasty. Many surgeons prefer not to prescribe chemoprophylaxis after arthroplasty. PMID- 11440001 TI - 10th International Conference on Human Retrovirology: HTLV and Related Viruses. June 25-29, 2001, Dublin, Ireland. Scientific program and abstracts. PMID- 11440002 TI - Patterns and diagnostic value of cardiac troponin I vs. troponin T and CKMB after OPCAB surgery. AB - BACKGROUND: Cardiac troponin I (cTnI) has been shown to be a specific marker for myocardial injury in cardiac surgery. The object of this prospective study was to determine the patterns and kinetic and diagnostic value of cTnI, cardiac troponin T (cTnT), and creatine kinase MB (CKMB) activity after minimally invasive coronary revascularization using an octopus device on the beating heart (OPCAB). METHODS: 48 patients (33 male/15 female, mean age 68.3 +/- 8.7 years) underwent their first elective OPCAB surgery with median sternotomy without mortality. The mean number of grafts was 2.0 +/- 0.8 per patient. Preoperative mean ejection fraction was 56.6 % +/- 14.9%. CTnI and T levels, total creatine kinase (CK) and CK-MB activity in the serum were measured before operation, at arrival at the ICU, and 6, 12, 24, 48 and 120 hours afterward. Serial 12-lead ECGs were recorded preoperatively and at days 1, 2 and 5. The relationship between perioperative data and postoperative cTnI and cTnT levels and CKMB were statistically identified for all variables. RESULTS: The best cutoff value for cTnI was 8.35 micrograms/l. The patients were grouped by the ECG findings and maximal slopes of cTnI postoperatively (group I: unchanged ECG and cTnI < 8.35 micrograms/l, n = 38; group II: unchanged ECG and cTnI > 8.35 micrograms/l n = 6; group III: Q-wave in ECG and cTnI > 8.35 micrograms/l, n = 4). Baseline serum concentrations of cTnI were in the normal range, and significantly increased after surgery with a peak 24h after the operation. Maximal slopes of cTnI ranged in group II between 9.1 and 18.0 micrograms/l, and in group III between 35.9 and 88.8 micrograms/l. There was strong concordance between maximum cTnI, cTnT (p < 0.0001) and CK-MB levels (p = 0.003). First cTnI levels immediately post-op correlated with the maximum cTnI levels during the postoperative course (p = 0.009). CONCLUSIONS: CTnI after minimal invasive surgery shows a characteristic pattern with a maximum at 24h after the operation. The measurement of postoperative biochemical marker concentrations, specially cTnI, reflects myocardial injury incurred during the procedure. It is an accurate method for confirming or excluding a perioperative myocardial injury diagnosis after OPCAB surgery. PMID- 11440003 TI - H&HN honored. Magazine of the Year. PMID- 11440004 TI - Staffing watch. PMID- 11440005 TI - Why docs oppose a deal. PMID- 11440006 TI - Medication safety issue brief 6. Finding and using resources. AB - It's one thing to understand the theories of patient safety. But it's another thing to carry them out. One helpful method is to use a question-and-answer style tool to examine how your organization measures up. Several tools are widely distributed by organizations like the AHA. There's plenty of other advice available, much of it just a mouse click away (see Action Agenda on flip side of this brief). PMID- 11440007 TI - A commitment to safety. PMID- 11440008 TI - So, you want to be a rural CEO. Why? PMID- 11440009 TI - [Spring Meeting of Internal Medicine. Lille, France, 27-28 March 2001. Proceedings and abstracts]. PMID- 11440010 TI - [Diagnostic test in clinical nutrition for medical students. Test results show that training in clinical nutrition must be improved]. AB - A national core curriculum in clinical nutrition was approved by the Section for Nutrition in the Swedish Society of Medicine in 1995. Here we report on the results of an anonymous diagnostic test based on this core curriculum in clinical nutrition, administered to medical students at the end of medical school. The test was the same for students in Linkoping, Lund and Stockholm. Only 42% of the participants obtained an acceptable test result, with the score in Lund being significantly lower than those in Linkoping and Stockholm. We compare the results with a similar test administered in Stockholm in 1996, and discuss current developmental work in clinical nutrition being done in all the medical faculties in Sweden. PMID- 11440011 TI - Diabetes UK. Annual professional meeting. 4-6 April 2001, Glasgow, UK. Abstracts and posters. PMID- 11440012 TI - [Editorial]. PMID- 11440013 TI - From the Centers for Disease Control and Prevention. The global HIV and AIDS epidemic, 2001. PMID- 11440015 TI - JAMA patient page. Genital herpes. PMID- 11440014 TI - From the Centers for Disease Control and Prevention. HIV and AIDS--United States, 1981-2000. PMID- 11440016 TI - British Society of Audiology short papers meeting on experimental studies of hearing and deafness. University of Keele, 21-22 September 2000. Abstracts. PMID- 11440017 TI - Internal transport barrier triggered by neoclassical transport in W7-AS. AB - The three-dimensional magnetic configuration of a stellarator offers two specific mechanisms for a transition to improved particle and energy confinement. One route goes through the so-called electron-root confinement regime, which leads to a reduction of neoclassical transport via strong radial electric fields. In this Letter evidence for a second route is presented. It opens due to the layer of a strongly varying radial electric field which is present in the transitional region from neoclassical electron to ion-root confinement. This type of improvement acts on turbulent transport. PMID- 11440022 TI - 11th European Meeting on Hypertension. Milan, Italy, June 15-19, 2001. Abstracts. PMID- 11440024 TI - 673rd meeting, University of Bristol. Abstracts. PMID- 11440023 TI - Telemedicine 2001, 6th International Conference on the Medical Aspects of Telemedicine. Uppsala, Sweden, 18-21 June 2001. Abstracts. PMID- 11440025 TI - Retraction. PMID- 11440026 TI - Receptor imaging in breast carcinoma: future prospects. PMID- 11440027 TI - Technetium-99m sestamibi imaging to predict left ventricular ejection fraction outcome after revascularisation in patients with chronic coronary artery disease and left ventricular dysfunction: comparison between baseline and nitrate enhanced imaging. AB - Acceptance of technetium-99m sestamibi as a tracer of myocardial viability is growing, particularly when nitrate-enhanced imaging is used. However, few data are available on the ability of 99mTc-sestamibi to predict the evolution of global left ventricular ejection fraction (EF). The aim of this study was to examine the ability of resting and nitrate 99mTc-sestamibi single-photon emission tomography (SPET) to predict EF changes after revascularisation in patients who have chronic coronary artery disease with left ventricular dysfunction. Using baseline resting and nitrate 99mTc-sestamibi SPET, we studied 61 patients scheduled for revascularisation because of left ventricular dysfunction. EF was estimated using two-dimensional echocardiography before and after the intervention. A post-revascularisation improvement of > or =5 EF units was defined as significant. Using a 13-segment model, 99mTc-sestamibi activity was quantified and the nitrate-induced activity changes calculated. Three different criteria for detecting viability (defined as post-revascularisation reversible dysfunction) in asynergic segments were compared: (1) resting 99mTc-sestamibi activity > or =60%; (2) nitrate 99mTc-sestamibi activity > or =65%; and (3) nitrate-induced increase >+10% or nitrate-induced increase < or =+10% and nitrate activity > or =65%. EF increased significantly in 32 patients. The number of viable asynergic segments was significantly higher in these patients than in the remaining 29 subjects, and the difference was greater (P<0.0002) using definition (3) than using either baseline (P<0.002) or nitrate activity (P<0.0005). There was a significant relationship between EF changes and number of viable asynergic segments: Spearman R=0.38, P<0.005 using baseline; Spearman R=0.39, P<0.002 using nitrate activity; and Spearman R=0.55, P<0.000005 using definition (3). According to receiver operating characteristic (ROC) curve analysis, this last criterion achieved the best results (81% sensitivity, 69% specificity and 75% accuracy), with an area under the ROC curve of 0.838; this area was significantly larger than when using either baseline (0.744, P<0.02) or nitrate activity (0.747, P<0.005). 99mTc-sestamibi SPET appears able to predict the evolution of global left ventricular EF after revascularisation, thereby confirming the value of 99mTc-sestamibi as a tracer of myocardial viability. The combination of baseline resting and nitrate imaging seems to significantly improve the diagnostic accuracy of 99mTc-sestamibi SPET for this particular purpose. PMID- 11440028 TI - A prospective blinded evaluation of exercise thallium-201 SPET in patients with suspected chronic exertional compartment syndrome of the leg. AB - This study compared the quantitative and qualitative results of leg thallium-201 single-photon emission tomography (SPET) imaging in patients with and without raised intracompartmental pressure associated with exercise-related leg pain. The purpose of this study was to clarify the aetiology of chronic exertional compartment syndrome (CECS), and to investigate the diagnostic applications of 201Tl SPET in CECS. Thirty-four study participants underwent compartment pressure testing (CPT) between March and August 2000. There were 25 positive CPT results (patient group), and nine negative CPT results (control group). All 34 participants underwent scintigraphy. Quantitative and qualitative assessments were performed for the anterolateral and deep posterior compartments of the lower leg. There was no significant difference in either quantitative or qualitative assessments of perfusion between those compartments with and those without CECS. In contrast, a marked effect of exercise type upon compartment perfusion pattern was noted. Results of this study indicate that there is no compartment perfusion deficit in those patients with raised intracompartmental pressure associated with CECS, and suggest a non-ischaemic basis for the pain associated with CECS. They also suggest no role for exercise perfusion scintigraphy in the diagnosis of this syndrome. PMID- 11440029 TI - Advantage of delayed whole-body FDG-PET imaging for tumour detection. AB - Delayed imaging that coincides with the highest uptake of fluorine-18 fluorodeoxyglucose (FDG) by tumour may be advantageous in oncological positron emission tomography (PET), where delineation of metastasis from normal tissue background is important. In order to identify the better imaging protocol for tumour detection, whole-body FDG-PET images acquired at 1 h and 2 h after injection were evaluated in 22 subjects, with a post-injection transmission scan at 90 min for attenuation correction. After visual interpretation, tumour uptake [tumour standardised uptake ratio (SUR)], normal tissue uptake (normal SUR) and tumour to background contrast (tumour SUR/normal tissue SUR) were evaluated in the images acquired at 1 h and at 2 h. Most malignant lesions, including primary lung cancer, metastatic mediastinal lymph nodes and lymphoma lesions, showed higher FDG uptake at 2 h than at 1 h. By contrast, benign lesions, with the exception of sarcoidosis, showed lower uptake of FDG at 2 h than at 1 h. Among normal tissues, the kidney, liver, mediastinum, lung, upper abdomen and left abdomen showed significant falls in FDG uptake from 1 h to 2 h. The lower abdomen, right abdomen and muscles (shoulder and thigh) showed no significant changes. Consequently, malignant lesions of the lung, mediastinum and upper abdomen showed significant increases in tumour to background contrast from 1 to 2 h. Three lesions (two lung cancers and a malignant lymphoma) that were equivocal on 1-h images became evident on 2-h images, changing the results of interpretation. All other malignant lesions were detected on 1-h images, but were clearer, with higher contrast, on 2-h images. Lesion-based sensitivity was improved from 92% (49/53) to 98% (52/53), and patient-based sensitivity from 78% (14/18) to 94% (17/18). It is concluded that delayed whole-body FDG-PET imaging is a better and more reliable imaging protocol for tumour detection. PMID- 11440030 TI - Quantitative evaluation of skeletal tumours with dynamic FDG PET: SUV in comparison to Patlak analysis. AB - This study was carried out to evaluate bone lesions using fluorine-18 fluorodeoxyglucose positron emission tomography (FDG PET) and to explore whether dynamic and quantitative PET data may help to differentiate benign lesions from malignant masses. Forty patients with primary bone lesions were studied. The final diagnosis was confirmed by histopathology. A 60-min dynamic FDG PET acquisition was undertaken in all subjects. From the dynamic PET images, indices such as the average and maximal standardised uptake values (SUVs), the tumour SUV to-muscle SUV ratio (T/M) and the SUV at 60 min-to-SUV at 30 min ratio (averSUV60/30 min and maxSUV60/30 min) were produced. Patlak graphical analysis was used to obtain the influx constant (Ki), and the metabolic rate of FDG (MRFDG) was calculated. Based on the receiver operator characteristic curve, the sensitivity and specificity for each parameter in differentiating between malignant and benign lesions were evaluated. The histological results revealed 21 malignant tumours and 19 benign lesions in this group. The MRFDG and SUV indices in malignant lesions were significantly higher than those in benign lesions. However, each index showed a considerable overlap between benign and malignant lesions. Average SUV correlated positively with MRFDG (r=0.67). When a cut-off of 1.8 average SUV was used, the sensitivity and specificity for discrimination of malignancy from benign disease were 85% and 82.4%, respectively. MRFDG showed a similar sensitivity (82.4%) and a better specificity (92.9%). A combination consisting of a cut-off of average SUV (1.8) and averSUV60/30 min (1.1) resulted in an improvement of specificity to 93.3%, with a small reduction in sensitivity (81.3%) as compared with exclusive use of SUV. The results of this study indicate that a detectable difference in glucose metabolism exists between malignant and benign skeletal lesions. The static FDG uptake indices alone may not enable adequate differentiation between benign and malignant lesions. Quantitative dynamic imaging may provide more helpful information, but will not permit a definite diagnosis. The use of uptake indices may represent an alternative and interesting approach to the evaluation of bone lesions. PMID- 11440031 TI - 18F-FDG PET and 99mTc-sestamibi scintimammography for monitoring breast cancer response to neoadjuvant chemotherapy: a comparative study. AB - Presurgical neoadjuvant chemotherapy has shown promise in the treatment of locally advanced breast carcinoma (LABC). Response assessment by clinical examination and mammography is difficult. This study evaluated and compared fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG-PET) and technetium-99m sestamibi scintimammography (SMM) as potential methods for the early assessment of tumour response to neoadjuvant chemotherapy in patients with LABC. Seven patients underwent PET and SMM [planar and single-photon emission tomography (SPET)] before beginning chemotherapy, after the first and second cycles of chemotherapy and after completing chemotherapy prior to surgery. PET and SMM results were evaluated visually and semi-quantitatively by calculating standardised uptake values (SUV) and tumour/lung ratios in the initial and subsequent studies. The findings were correlated with the initial clinical and mammographic findings and the final histopathological diagnoses. There was a highly significant correlation between SUVmean, SUVmax and the tumour/lung ratio determined with SMM-SPET in the studies performed before and during neoadjuvant chemotherapy. All three patients with complete remission showed decreasing FDG and sestamibi uptake as early as 8 days after therapy. In the presurgical study, increased sestamibi and FDG uptake was no longer evident. Three patients had partial remission with clearly reduced but persisting focal FDG and sestamibi uptake after neoadjuvant therapy. One patient who did not respond to therapy had unchanged intense tracer uptake during chemotherapy that was evident with both techniques. An early decline in glucose metabolism or sestamibi uptake 8 days after beginning therapy did not necessarily predict complete tumour remission in the further course of chemotherapy. On the other hand, increased tracer uptake after the first cycle did not exclude a partial tumour response. After the second chemotherapeutic cycle both techniques were able to distinguish between complete and partial/no response. There was a good correlation between preoperative FDG and sestamibi uptake and the histopathologically determined tumour size. However, small residual invasive tumours in patients with clinically complete remission could not be visualised with either technique. The preliminary data demonstrate that sestamibi SMM is as useful as FDG-PET for the monitoring of tumour response to neoadjuvant chemotherapy. PMID- 11440032 TI - Comparison of [18F]FHPG and [124/125I]FIAU for imaging herpes simplex virus type 1 thymidine kinase gene expression. AB - Various radiotracers based on uracil nucleosides (e.g. [124I]2'-fluoro-2'-deoxy-5 iodo-1-beta-D-arabinofuranosyluracil, [124I]FIAU) and acycloguanosine derivatives (e.g. [18F]9-[(3-fluoro-1-hydroxy-2-propoxy) methyl] guanine, [18F]FHPG) have been proposed for the non-invasive imaging of herpes simplex virus type 1 thymidine kinase (HSV1-tk) reporter gene expression. However, these radiotracers have been evaluated in different in vitro and in vivo models, precluding a direct comparison. Therefore, we directly compared [18F]FHPG and radioiodinated FIAU to assess their potential for PET imaging of transgene expression. The uptake of [125I]FIAU, [18F]FHPG and [3H]acyclovir was determined in vitro using four different HSV1-tk expressing cell lines and their respective negative controls. The in vitro tracer uptake was generally low in non-transduced parental cell lines. In HSV1-tk expressing cells, [3H]acyclovir showed approximately a twofold higher tracer accumulation, the [18F]FHPG uptake increased by about sixfold and the [125I]FIAU accumulation increased by about 28-fold after 120-min incubation of T1115 human glioblastoma cells. Similar results were found in the other cell lines. In addition, biodistribution and positron emission tomography (PET) studies with [18F]FHPG and [124/125I]FIAU were carried out in tumour-bearing BALB/c mice. Significantly higher specific accumulation of radioactivity was found for [125I]FIAU compared with [18F]FHPG. The ratio of specific tracer accumulation between [125I]FIAU and [18F]FHPG increased from 21 (30 min p.i.) to 119 (4 h p.i.). PET imaging, using [124I]FIAU, clearly visualised and delineated HSV1-tk expressing tumours, whereas only a negligible uptake of [18F]FHPG was observed. This study demonstrated that in vitro and in vivo, the radioiodinated uracil nucleoside FIAU has a significantly higher specific accumulation than the acycloguanosine derivative [18F]FHPG. This suggests that [124I]FIAU should be the preferred reporter probe for PET imaging of HSV1-tk gene expression. Thus, further attempts to develop suitable PET tracers for the assessment of HSV1-tk gene expression should also focus on 18F-labelled uracil derivatives. PMID- 11440033 TI - Effects of insulin and glucose loading on FDG uptake in experimental malignant tumours and inflammatory lesions. AB - Fluorine-18 2-deoxy-2-fluoro-D-glucose (FDG) accumulation in tumours has been well investigated, but much less is known regarding FDG accumulation in inflammatory lesions. In this study, we determined the effects of hypo- and hyperglycaemia on FDG uptake in inflammatory lesions of infectious and non infectious origin and compared them with those in malignant tumours in rats, to provide a biological basis for differentiating malignant lesions from benign lesions by means of FDG-PET. Rats were inoculated with a suspension of allogenic hepatoma cells (KDH-8) or Staphylococcus aureus, or with turpentine oil into the left calf muscle. Two weeks after KDH-8 inoculation and 1 week after S. aureus and turpentine oil inoculations, the rats were divided into three subgroups: insulin-loaded (2 U/kg body weight, i.p.), glucose-loaded (1.2 g/kg body weight, p.o.) and control groups. Radioactivity in tissues was determined 1 h after i.v. injection of FDG. Intraperitoneal injection of insulin and oral administration of glucose induced hypoglycaemia and hyperglycaemia, respectively. In the control animals, tumours showed a level of FDG uptake which was 2.2 and 3.0 times higher than the levels in the inflammatory lesions induced by S. aureus and turpentine oil, respectively (P<0.0001). There was no significant difference in the level of FDG uptake between the two inflammatory lesions of infectious and non-infectious origin. Insulin loading significantly decreased the level of FDG uptake in tumours and in both types of inflammatory lesion to approximately one-half of the control values (P=0.001 in the tumour group and P<0.0001 in the two inflammatory lesion groups). In the glucose-loaded group, the level of FDG uptake in both types of inflammatory lesion decreased significantly to 50%-61% of the control value (P=0.0002 in the S. aureus group and P<0.0001 in the turpetine group), while the tumour uptake did not decrease significantly (86% of the control value) (P=NS). It is concluded that FDG uptake in both types of inflammatory lesion was significantly impaired in rats with hyperglycaemia induced by glucose loading, while tumour uptake of FDG was not significantly affected. These results indicate that glucose loading has greater effects on FDG uptake in inflammatory lesions than in tumours, providing a biological basis for differentiation of malignant lesions from benign lesions by FDG-PET in a clinical setting. PMID- 11440034 TI - The value of oblique pinhole images in pre-operative localisation with 99mTc-MIBI for primary hyperparathyroidism. AB - Technetium-99m labelled 2-methoxyisobutylisonitrile (MIBI) has been extensively utilised for preoperative localisation of parathyroid adenomas. Imaging techniques have varied widely, with many centres not performing routine oblique images; thus this study aimed to examine the value of routine oblique pinhole imaging. Ninety-two patients underwent pre-operative 99mTc-MIBI imaging including early and delayed anterior oblique pinhole images in addition to standard anterior pinhole images and a thyroid study prior to surgery for primary hyperparathyroidism. These studies were reviewed blindly comparing anterior and oblique images and anterior images only in relation to surgical findings. Of the 92 patients, 83 were found to have 86 parathyroid adenomas or parathyroid adenoma/hyperplasia at surgery. When compared to anterior images only, oblique views improved overall sensitivity from 76% to 88% (P<0.05), correctly localised 11 more adenomas than anterior images alone (13%) and improved the confidence of interpretation in 17 patients (20%). In conclusion, routine oblique pinhole views result in greater sensitivity and reporter confidence in pre-operative parathyroid localisation with 99mTc-MIBI. PMID- 11440035 TI - Targeting of liver tumour in rats by selective delivery of holmium-166 loaded microspheres: a biodistribution study. AB - Intra-arterial administration of beta-emitting particles that become trapped in the vascular bed of a tumour and remain there while delivering high doses, represents a unique approach in the treatment of both primary and metastatic liver tumours. Studies on selective internal radiation therapy of colorectal liver metastases using yttrium-90 glass microspheres have shown encouraging results. This study describes the biodistribution of 40-microm poly lactic acid microspheres loaded with radioactive holmium-166, after intra-arterial administration into the hepatic artery of rats with implanted liver tumours. Radioactivity measurements showed >95% retention of injected activity in the liver and its resident tumour. The average activity detected in other tissues was < or =0.1%ID/g, with incidental exceptions in the lungs and stomach. Very little 166Ho activity was detected in kidneys (<0.1%ID/g), thereby indicating the stability of the microspheres in vivo. Tumour targeting was very effective, with a mean tumour to liver ratio of 6. 1+/-2.9 for rats with tumour (n=15) versus 0.7+/-0.5 for control rats (n=6; P<0.001). These ratios were not significantly affected by the use of adrenaline. Histological analysis showed that five times as many large (>10) and medium-sized (4-9) clusters of microspheres were present within tumour and peritumoural tissue, compared with normal liver. Single microspheres were equally dispersed throughout the tumour, as well as normal liver parenchyma. PMID- 11440036 TI - Improved response of colon cancer xenografts to radioimmunotherapy with pentoxifylline treatment. AB - A methylxanthine, pentoxifylline (PTX), has the potential to improve tumour microcirculation and oxygenation in vivo. We aimed to determine whether this agent would enhance the response of tumours to experimental radioimmunotherapy (RIT). Balb/c nu/nu mice with xenografts of LS180 human colon cancer were treated with 4.63 MBq of 131I-A7 anti-colorectal monoclonal antibody. A dose of 50 mg/kg of PTX was administered i.p. immediately after the 131I-A7 injection and daily thereafter for 7 days. The effect of PTX administration on 131I-A7 targeting in tumours was assessed with biodistribution and radioluminography on day 2. Intratumoural pO2 was measured with microelectrodes. The administration of PTX alone did not suppress tumour growth, but the efficacy of RIT with 131I-A7 was significantly improved by PTX: tumour volumes on day 15, relative to the initial volume, were 16.8+/-3.60 in the nontreated controls, 13.9+/-2.17 with PTX, 3.43+/ 0.44 with RIT, and 1.86+/-0.59 with RIT+PTX (P<0.05). PTX administration did not alter the biodistribution or intratumoural distribution of 131I-A7. However, intratumoural pO2 was significantly improved by PTX administration: 16.9+/-9.75 mmHg in control tumours versus 25.6+/-11.3 mmHg in PTX-treated tumours (P<0.01). These results indicate that PTX-induced radiosensitisation of tumour cells due to better oxygenation is responsible for the better RIT outcomes, because the net radiation absorbed dose to the tumours did not appear to be changed. PMID- 11440037 TI - In vivo evaluation of 111In-DTPA-N-TIMP-2 in Kaposi sarcoma associated with HIV infection. AB - Matrix metalloproteinases are the major agents responsible for the degradation of extracellular matrix and are produced at high levels by transformed and tumour cells, where they participate in the metastatic process by allowing local invasion. They are also more active at sites of new normal growth and angiogenesis. In the early stages of Kaposi sarcoma (KS), in vitro studies have demonstrated that vascular invasion can be inhibited by inhibitors of matrix metalloproteinases. Imaging of visceral and cutaneous KS presents a problem and therefore the potential use of a labelled inhibitor of metalloproteinases, N-TIMP 2, with indium-111 was thought to present a possible imaging tool. The biokinetics, dosimetry and potential for imaging with 111In-DTPA-N-TIMP-2 were assessed in five patients with HIV infection and KS. Between 103.1 and 108.0 MBq of this agent was injected into each patient, and the dynamic uptake over the kidneys was assessed, whole body scans were performed and blood samples were obtained. The clearance from the blood was rapid, with a first component half time of 16.6+/-3.4 min and a second component half-time of 9.68+/-2.68 h. Two out of five patients experienced minor shivering but one of these patients was generally unwell before the study. The last three patients had no such problems. The tracer distributed predominantly to the kidneys and did not localise in other tissues. No KS lesions were clearly identified. 111In-DTPA-N-TIMP-2 can be successfully prepared and administered to patients safely, with a biodistribution and dosimetry which would allow its use as an imaging tracer. It is unlikely to be of use for imaging KS, but may have a role in other tumours that produce matrix metalloproteinases. PMID- 11440038 TI - The clinical value of 18F-FDG detection with a dual-head coincidence camera: a review. AB - Positron emission tomography (PET) has evolved into a technique that can accurately determine the distribution of positron-emitting radionuclides. The addition of a coincidence detection mode to a standard dual-head detector system has resulted in the option of single-photon and annihilation coincidence detection. This new device for imaging fluorine-18 2-fluoro-2-deoxy-D-glucose (18F-FDG) accumulation in neoplasms became commercially available in 1994. Besides conventional low-energy imaging in the collimated single-photon mode, it offers a relatively inexpensive opportunity to perform uncollimated PET by switching to the coincidence acquisition mode. This review summarises the clinical value of 18F-FDG detection with a dual-head coincidence camera in oncology. The results are compared with the overall results obtained using dedicated PET scanners. With respect to head and neck tumours, 18F-FDG coincidence mode gamma camera imaging (CGI) yields results that are in agreement with those obtained with dedicated PET scanners. With regard to other malignancies, such as lung cancer, lymphoma and brain tumours, data in the literature are too scarce to draw any definite conclusions. In general, the results of 18F-FDG CGI in tumours >15 mm seem to be comparable to those obtained with dedicated PET scanners, whereas in tumours <15 mm, the relative sensitivity of 18F-FDG CGI is approximately 80%. Using attenuation correction, the diagnostic yield of 18F-FDG CGI may increase. However, further clinical investigation is required to definitely establish its value in staging primary disease, therapy monitoring and assessment of tumour recurrence in clinical oncology. PMID- 11440039 TI - Concerns about 99mTc-labelled ciprofloxacin for infection detection. PMID- 11440040 TI - AIDS strategy 2000. PMID- 11440041 TI - The price of a research subject. PMID- 11440042 TI - Anabolic steroids, brain and behaviour. PMID- 11440043 TI - Exposure to passive smoke in a sample of children in North Western Ireland. AB - Passive smoke causes significant detrimental effects on the health of both children and adults. This study involved distributing a questionnaire to a sample of 265 school children aged between 12 and 17 years in North West Ireland. The aims were to establish the prevalence of passive smoking and to discern whether exposure to passive smoke takes place within or outside the home. Of the 262 valid questionnaires, 188 pupils (72%) classified themselves as non-smokers. Sixty-five pupils (25%) described their mothers as current smokers whilst sixty one (23%) described their fathers as current smokers. Eighty-five pupils (32%) were exposed to smoke at home and 195 (74%) were exposed to smoke in the wider community, whilst 69 (26%) were exposed to smoke both at home and in the community. Two hundred and eleven pupils (81%) were therefore exposed to passive smoke at some location. Further measures are required to reduce the level of passive smoking. It is important that interventions should restrict smoking in public places in addition to promoting a smoke-free environment in the home. PMID- 11440044 TI - Patient satisfaction with general practice in Ireland. AB - Patient satisfaction in healthcare is gaining widespread recognition as a measure of ascertaining how well health services are being delivered. An analysis of the non-clinical elements of general practice in Ireland using a patient satisfaction questionnaire (PSQ) presents data collected from 2,167 patients to examine patient satisfaction. The level of satisfaction reported in the PSQ was 74.5% (n=1614) with dissatisfaction at 14.5% (n=314). The areas of general practice with the highest level of reported satisfaction are communication 83.7% (n=1814), general satisfaction 81.2% (n=1760), access 74.4% (n=1612) and premises 75% (n=1625), whereas, appointments and availability shows a considerably lower level of satisfaction at 58.6% (n=1270). Further analysis was undertaken to examine how patients' characteristics influenced their responses. The age, educational level, gender and public/private status of the patient all influenced satisfaction levels. It was concluded that if PSQ's are to contribute to developing service delivery and good practice they must take into account the differing characteristics of patients. PMID- 11440045 TI - Irish orthopaedic patients' attitudes to blood transfusion. AB - This study was designed in response to perceived increased public anxiety about the risks of allogenic blood transfusion. A questionnaire was completed by 203 patients attending an elective orthopaedic hospital. The questionnaire examined patients' factual knowledge about various aspects of transfusion, their attitudes to blood transfusion and confidence in the safety of blood products. Despite recent publicity, only 124 patients (61%) regarded blood transfusion as a risk for hepatitis transmission while 172 patients (85%) knew of the risk of blood transmission of HIV. 22 patients (11%) reported that they would have no concerns about receiving a blood transfusion, while at the other extreme, 9 (5%) would not be willing to receive blood under any circumstances. 162 patients (80%) felt that allogenic blood transfusion was either entirely safe or that there was an acceptable small risk associated with it. A majority of responders (109 patients, 54%) reported they would favour autologous transfusion if available. Current practices and alternatives to traditional allogenic blood transfusion are discussed in the context of elective orthopaedic surgery. PMID- 11440046 TI - Antenatal hepatitis B screening - is there a need for a national policy? AB - Routine antenatal testing for hepatitis B carriage with maternal consent was introduced at the Rotunda in January 1998. The uptake of testing has been excellent; 99.98% of women presenting for antenatal care accepted hepatitis B (HBV) screening in the 30-months from January 1998 through June 2000. The prevalence of HBV carriage was 0.35% (58 pregnancies of 16,222 tested) increasing from 0.25% in 1998 (16 of 6227) to 0.45% in the first six months of 2000 (16 of 3484). Fifty-five women had 58 pregnancies (three women had two pregnancies). Two of these were e-antigen positive. HBV carrier status was previously unknown in 48 (87%). Two additional women had acute HBV infection in pregnancy. Forty-five infants have been born to mothers included in this screening programme. Audit of infant outcome reveals excellent compliance with immunisation and follow-up: 29 (64%) have completed the 3 dose HBV vaccination schedule to date. Thirteen infants (31%) are still attending; three are lost to follow-up including one whose family has emigrated. Routine antenatal screening for hepatitis B carriage is cost-effective and should be considered a standard of care in maternity practice. PMID- 11440047 TI - Gallstones and laparoscopic cholecystectomy in hepatitis C patients. AB - Our aims were to determine the incidence of gallstones in a cohort of patients infected with hepatitis C for 20 years, and to analyse the outcome of all patients infected with hepatitis C undergoing laparoscopic cholecystectomy. A hepatitis C screening programme in place in Ireland since 1994 identified 965 patients with hepatitis C antibodies out of 62,667 patients screened. The hepatology unit of Cork University Hospital has 318 patients with hepatitis C. Of patients identified by screening, 201 were post partum women infected via contaminated Anti-D immunoglobulin administered in 1977. Thirty-five (17.4%) of two hundred and one patients with hepatitis C since 1977 had developed gallstones after twenty years. A total of 34 patients with hepatitis C underwent laparoscopic cholecystectomy. One patient required conversion to open cholecystectomy. There were no complications and no mortality. There was a low rate of cirrhosis (11%) on examining liver histology. The incidence of gallstones in a cohort of patients infected with hepatitis C for twenty years approximates to that of the general population. The low rate of cirrhosis in this group may be related to a low consumption of alcohol. Laparoscopic cholecystectomy is a safe procedure in patients with mild chronic liver disease caused by hepatitis C. PMID- 11440048 TI - Sigmoid sinus thrombosis: an old foe revisited. AB - Sigmoid sinus thrombosis (SST) has become increasingly uncommon. In the pre antibiotic era this condition had a mortality rate of over 90%.1 A high index of suspicion is required to make the diagnosis. We present a rare case of sigmoid sinus thrombosis secondary to mastoiditis, which illustrates the problems of delayed diagnosis. This report highlights the importance of rapid diagnosis and early surgical intervention. We emphasis the need for scanning and otolaryngology referral in all cases of middle ear disease associated with pain or vertigo which does not resolve rapidly on appropriate antibiotic therapy. PMID- 11440049 TI - Orthopaedic workload of county Mayo. PMID- 11440050 TI - Cardiac enzyme levels in patients receiving oral neuroleptics. PMID- 11440051 TI - Cannabis. PMID- 11440052 TI - Historical background of antiseptic surgery. PMID- 11440053 TI - The erosion of citizenship. AB - The Marshallian paradigm of social citizenship has been eroded because the social and economic conditions that supported postwar British welfare consensus have been transformed by economic and technological change. This article argues that effective entitlement was based on participation in work, war and reproduction, resulting in three types of social identity: worker-citizens, warrior-citizens and parent-citizens. The casualization of labour and the technological development of war have eroded work and war as routes to active citizenship. Social participation through reproduction remains important, despite massive changes to marriage and family as institutions. In fact the growth of new reproductive technologies have reinforced the normative dominance of marriage as a social relation. These rights of reproduction are described as 'reproductive citizenship'. The article also considers the role of voluntary associations in Third-Way strategies as sources of social cohesion in societies where social capital is in decline, and argues that the voluntary sector is increasingly driven by an economic logic of accumulation. With the erosion of national citizenship, Marshall's three forms of rights (legal, political and social) have been augmented by rights that are global, namely environmental, aboriginal and cultural rights. These are driven by global concerns about the relationship between environment, community and body such that the quest for social security has been replaced by concerns for ontological security. PMID- 11440054 TI - Tightening the net: children, community, and control. AB - The recent move to revitalize social democracy in the UK under the New Labour government, explored by Giddens as 'the Third Way', embraces many of Etzioni's ideas on communitarianism. The principles that emerge from these political philosophies, such as the involvement of local communities in policy consultations and implementation, have largely been welcomed as a reflection of the aim of revitalizing civic society in the context of a range of social policies. It is argued, however, that for children, contrary to this general trend, many of these policies represent attempts to increase the social control of children. Their effect has been to restrict children's agency and their rights, rather than to increase their participation as citizens, and thus,in spite of the requirements of the UN Convention on the Rights of the Child, children continue to be marginalized. PMID- 11440055 TI - Analysing patterns of religious participation in post-communist Eastern Europe. AB - It is generally thought that processes of modernization generic to industrialized societies have resulted in a process of secularization with respect to conventional religious participation and observance in most Western countries. It is not at all clear, however, whether the post-communist societies of Eastern Europe have followed this pattern. In this we paper we examine whether levels of religiosity in ten post-communist societies--five generally Catholic in orientation and five Orthodox--are consistent with secularization theory, or whether instead they display, as some have suggested, the impact of seven decades of atheistic communism followed by a recent resurgence among the young. For this purpose we examine denominational membership and church attendance using descriptive and multivariate analysis of large-scale national sample surveys conducted in the mid-1990s. We find that age and educational differences in participation rates follow patterns expected on the basis of secularization theory with no evidence of resurgence among younger groups. Also, however, Catholic participation rates are significantly higher than Orthodox ones, indicating the importance of denomination in understanding patterns of religiosity in the post-communist context. PMID- 11440056 TI - Family business in Russia: the path to middle class? AB - The paper seeks to explore via a series of interview-based case studies aspects of the emergence of an entrepreneurial middle-class in Russia. The paper notes the origins of those studied in the professional or highly skilled workers in the former Soviet Union. The paper reveals the complexity and fragility of the circumstances of these entrepreneurs and suggests that commentary in both Russia and the West that pins its hopes for social stability on the emergence of a new property owning middle class in Russia are, at best, premature. PMID- 11440057 TI - Moral panic versus the risk society: the implications of the changing sites of social anxiety. AB - This paper compares moral panic with the potential political catastrophes of a risk society. The aim of the comparison is threefold: 1. to establish the position of risk society threats alongside more conventional moral panics; 2. to examine the conceptual shifts that accompany the new types of threats; and 3. to outline the changing research agenda. The paper suggests that as new sites of social anxiety have emerged around environmental, nuclear, chemical and medical threats, the questions motivating moral panic research have lost much of their utility. Conceptually, it examines how the roulette dynamics of the risk society accidents expose hidden institutional violations that redound into 'hot potatoes' that are passed among and fumbled by various actors. Changing conceptions of folk devils, claims making activities, and of a safety are also discussed. PMID- 11440058 TI - BSE as an organizational construction: a case study on the globalization of risk. AB - This article examines the BSE problem as an example of the 'globalization of risk'. In order to determine whether the 'globalization of risk' is a social construction depending on the context, the paper emphasizes the particular role of organizations. It makes an empirical comparison of the BSE-related risk constructions of five business associations in the German meat industry sector. The results show that the associations construct the risk in close relation to their horizons of globalization, thereby reflecting provision problems, which the companies they are representing face. While the main organizational domains in the sector tried to cope with the risk problem by different means of local market 'closure', one association, founded in reaction to the BSE problem, took over a 'reflexive' role with regard to the emerging risk communication on BSE in Germany. PMID- 11440059 TI - The authority of complexity. AB - The assertion about the unique 'complexity' or the peculiarly intricate character of social phenomena has, at least within sociology, a long, venerable and virtually uncontested tradition. At the turn of the last century, classical social theorists, for example, Georg Simmel and Emile Durkheim, made prominent and repeated reference to this attribute of the subject matter of sociology and the degree to which it complicates, even inhibits the develop and application of social scientific knowledge. Our paper explores the origins, the basis and the consequences of this assertion and asks in particular whether the classic complexity assertion still deserves to be invoked in analyses that ask about the production and the utilization of social scientific knowledge in modern society. We present John Maynard Keynes' economic theory and its practical applications as an illustration. We conclude that the practical value of social scientific knowledge is not dependent on a faithful, in the sense of complete, representation of social reality. Instead, social scientific knowledge that wants to optimize its practicality has to attend and attach itself to elements of social situations that can be altered or are actionable. PMID- 11440060 TI - Cross-national changes in time-use: some sociological (hi)stories re-examined. AB - The paper addresses some macro-sociological questions about changes in broad categories of time-use. The focus is on large-scale cross-national time trends from developed countries in paid and unpaid work, and leisure. Reference is made to some well-known sociological and historical accounts of such change, and to the fact that time-use diary data has only relatively recently become available for analysing trends over time. The data used are drawn from a comparative cross time data archive held by the Institute for Social and Economic Research at Essex University, comprising successive time-use diary surveys from a range of industrialized countries collected from the 1960s to the 1990s. The time use evidence suggests relative stability in the balance between work and leisure time over the period covered by the analyses. Some alternative explanations are advanced for why there seems to be a gap between this evidence and, on the one hand, the burgeoning literature in both academic and popular media addressing the 'time famine' and, on the other, people's professed experience of what is happening to their time. PMID- 11440061 TI - Bringing health care information to the deaf community. AB - BACKGROUND: The Deaf community reports limited access to health promotion information and care. Literature review, key informant interviews, and focus groups generated a clearer understanding of the community. Health care providers, educators, and policymakers could improve medical care to the Deaf community by: 1) better understanding its culture and language; 2) creating more health education programs specifically for the Deaf community; 3) developing opportunities for more deaf people and American Sign Language (ASL) users to enter the health professions; and 4) creating incentives for hearing health care providers to become ASL proficient. PMID- 11440062 TI - Instruments to measure cancer management knowledge of rural health care providers. AB - BACKGROUND: Instruments to measure cancer management knowledge of rural physicians, nurses, and pharmacists were needed to evaluate the effect of an educational intervention. Since such instruments did not exist, the authors designed and validated a new instrument for each discipline. METHODS: The design and validation process for these instruments are described. RESULTS: These three instruments were shown to be practical and to have high content and construct validity. Content validation demonstrated that all items were rated as essential or useful by 90% or more of the respondents. Construct validation show highly significant differences in mean scores among several levels of learners and practitioners as expected. CONCLUSIONS: These instruments may be useful to other investigators for measuring cancer management knowledge of rural physicians, nurses, and pharmacists. PMID- 11440063 TI - Differences in cancer screening priorities between medical students and residents. PMID- 11440064 TI - Improving medical students' communication with limited-literacy patients: project development and implementation. AB - BACKGROUND: Physicians may not realize that some of their patients have limited reading abilities. The AMC Cancer Research Center and the Primary Care Curriculum at the University of Colorado collaborated to develop and implement a project to teach medical students to identify and effectively communicate with limited literacy patients regarding cancer screening. METHOD: The project incorporated two standardized-patient (SP) cases on cancer screening and literacy into the existing curriculum for second-year students. A small-group, interactive problem based learning (PBL) case on breast cancer screening and literacy introduced the concept of communicating about cancer screening and literacy issues. A clinical practice examination on colorectal cancer screening and literacy was given at the end of the year. RESULTS: The collaborative project proved successful in incorporating literacy and cancer screening into a medical school curriculum. Time spent training SPs to play the role of adults with limited literacy was very important, as was time spent pre-testing the cases. The PBL case effectively promoted group discussion of cancer screening and literacy. CONCLUSION: The project succeeded in making students aware of literacy and cancer screening issues. Outcome results are being analyzed. PMID- 11440065 TI - Breast cancer screening for primary care trainees: comparison of two teaching methods. AB - BACKGROUND: This study compared the efficacies of two methods of teaching breast cancer screening to primary care trainees. METHODS: Fifty-one nurse-practitioner students were assigned by class section and 47 medical residents by practice site to receive a lecture-demonstration class or individual/small-group instruction from a standardized patient. Prior to instruction and one year later, participants took a written test to assess knowledge and standardized patients evaluated their skills. RESULTS: Overall, the participants improved their breast cancer screening skills. CONCLUSION: The standardized patient teaching method was of greater benefit to the nurse-practitioner students. PMID- 11440066 TI - Mexico City physicians' awareness about cervical cancer prevention: implications for cancer screening. AB - BACKGROUND: In spite of an early cancer detection program (CCSP), Mexico has a mortality rate for cervical cancer of 16.5 per 100,000 women. METHOD: A cross sectional study of 330 physicians at the Mexico City General Hospital evaluated their knowledge of the CCSP, etiology, diagnostic alternatives, and treatment guidelines. Variance analysis was the statistical procedure used. Replies to a questionnaire about cervical cancer prevention awareness were scored on a scale from 1 to 9. RESULTS: According to the awareness scale, the global average classification was 4.4, with 50% of the physicians scoring 4 or less. There was no difference in the CCSP knowledge scores of gynecologists (mean 4.92, 95% CI 4.2-5.3), oncologists (mean 4.85, 95% CI 4.3-5.5), pathologists (mean 5.23, 95% CI 4.9-5.6), and those in other specialties (mean 4.29, 95% CI 4.2-5.0), p > 0.05. Many respondents attributed CCSP's lack of effectiveness to public apathy (68.12%). CONCLUSIONS: The effectiveness of the CCSP can be improved by educating health professionals if this education is combined with elimination of obstacles to its use. More information is needed to justify revising how doctors are educated in terms of not only quality of the training but also the contents of pre- and postgraduate training programs. PMID- 11440067 TI - Australian dentists' educational needs for smoking cessation counseling. AB - BACKGROUND: Australian dentists' continuing educational needs and their attitudes towards and self-reported practices related to smoking cessation counseling were examined. METHOD: Self-administered questionnaires were received from 149 dentists (83% response rate). RESULTS: Many dentists were aware that smoking is a risk factor for the development of oral cancer (n = 128, 86%). Most considered smoking cessation counseling to be part of their professional role (n = 105, 70%). However, few "always" asked about the smoking status of their patients (n = 21, 14%). The dentists' use of specific behavioral techniques known to assist patients to quit also was low. Furthermore, the dentists were as likely to use ineffective (advice to "cut down") as effective (advice to "quit") (p > 0.05) strategies. The respondents were significantly more interested in self-help pamphlets for their patients than in either evidence-based guidelines (McNemar's chi2 = 9.76, df = 1, p < 0.01) or a self-study module about smoking cessation (McNemar's chi2 = 42.0, df = 1, p < 0.001). CONCLUSIONS: Continuing education for dentists that combines skills training, patient materials, and epidemiology is likely to be acceptable and effective. PMID- 11440068 TI - Compliance with fecal occult blood test screening among low-income medical outpatients: a randomized controlled trial using a videotaped intervention. AB - BACKGROUND: Morbidity and mortality from colorectal cancer (CRC) are heightened among the socioeconomically disadvantaged. METHOD: A randomized controlled trial was conducted to evaluate the efficacy of a videotaped intervention using peer educators as well as a health professional to increase compliance with fecal occult blood test (FOBT) screening. Participants were 160 older individuals attending a medical outpatient clinic. Compliance with FOBT use was the dependent measure. Demographic variables, family history of CRC, viewing the videotape, perceived risk, self-efficacy, physician recommendation, knowledge about CRC screening, and intent to use the FOBT were independent measures. RESULTS: Approximately 41% of participants complied with FOBT screening. Significant relationships were found between intent and family history of CRC, viewing the video, perceived risk, self-efficacy, and CRC knowledge. However, none of these variables was significantly related to compliance with FOBT use. CONCLUSIONS: Although modest compliance rates were shown for both experimental and control groups, their compliance did not differ significantly. Further investigation of the impact of a video as part of an enhanced intervention program should be considered. PMID- 11440069 TI - Clinical trials in a rural population: professional education aspects. AB - BACKGROUND: While the majority of cancer patients in rural New Hampshire and Vermont are treated in community hospitals, few have entered clinical trials. This report describes a rural hospital consortium as a single Cancer and Leukemia Group B (CALGB) affiliate that used local cancer teams and itinerant oncologists to develop a clinical trials program. METHOD: Grafted onto an existing oncology outreach program, educational programs were developed to help identify patients and recruit them to cooperative group clinical trials. Outcomes included the number of patients accrued to clinical trials, and a comparison of the quality of audited research records with those of affiliated institutions of the CALGB. The consequences of the program were to measure changes in patterns of care of breast, prostate, colorectal, and lung cancers during the study period. These included diagnostic, staging, and treatment changes that occurred over time. RESULTS: 3.3% of incident cases were accrued to clinical trials during the study period, more often for breast and colorectal than for lung and prostate cancers. Reasons that were identified for low accrual were lack of clinical trials for the majority of cases, including the elderly. More than 65% of the patients in the outreach population were older than 65, compared with 50% at the cancer center. Patterns of care did change for breast and prostate cancers, but were similar to national trends. PMID- 11440070 TI - Use of modified OSCEs to verify technical skill performance and competency of lay caregivers. AB - BACKGROUND: The demands on lay caregivers of blood and marrow stem-cell transplant recipients are complex. Effective caregiver education that clearly delineates and demonstrates what needs to be learned is needed, as well as a method to evaluate caregiver's skill-based competencies. METHOD: A pilot study was conducted to determine the feasibility of adapting objective structured clinical examinations (OSCEs) for teaching and evaluation in lay caregiver educational programs. A modified OSCE, a performance-based assessment process, with seven practice stations for the care of tunneled central lines was developed and tested with 13 laypersons. Inter-rater techniques were used to establish accuracy and reproducibility of the examination. RESULTS: Reproducibility of station scenarios and accuracy in performance evaluation were supported by an overall kappa coefficient of 0.94, indicating excellent agreement. Participants' overall mean performance scores were 80% for knowledge and 94% for skills. CONCLUSION: The use of modified OSCEs for teaching and evaluating lay caregivers' competencies for complex care demands is feasible, accurate, and warrants further investigation. PMID- 11440071 TI - Cancer prevention in rural youth: teaching goals for health: the pilot. AB - BACKGROUND: The Goals for Health project is designed to change the cancer-related behaviors of tobacco use and dietary fat and fiber consumption. The intervention teaches health and life skills to rural, minority sixth and seventh graders in rural Virginia and New York. This article presents the results of the pilot. METHODS: Participants were 129 sixth graders at one rural middle school who were surveyed prior to and following delivery of the pilot sixth-grade intervention. RESULTS: Results include significant changes from pre- to post-intervention in several diet and smoking attitude and self-efficacy variables, dietary fat and fiber knowledge, high-fat snack consumption, and dietary fat scores. Multivariate analyses reveal important contributions of personal control over food choices and family and friend influence on change in dietary fat score from pre- to post intervention. CONCLUSIONS: These pilot program results suggest avenues for dietary and cancer prevention interventions in high-risk, rural adolescents. PMID- 11440072 TI - Synthesis and evaluation of the analgesic and antiinflammatory activities of O substituted salicylamides. AB - The present investigation deals with the synthesis of some new salicylamidoacetyl sulfonamides 3a,b, salicylamido ethylacetate 4, salicylamido acetic acid hydrazide 5, which is considered as the key intermediate for the synthesis of several series of new compounds such as salicylamido pyrazol 6 and pyrazolone 7. N-imido-derivatives 9, 10, 11, thiadiazole 13, oxadiazole 14, 15, Schiffs bases 16a-f. Cyclocondensation of Schiffs bases with thioglycolic acid gave thiazolidinone 18a-c while with acetylchloride afforded azitidinones 19a-c and with acetic anhydride gave 1,4-benzoxazepine-3,5-dione. Some of the compounds were tested for their analgesic and antiinflammatory activities as well as ulcerogenic effects. Some derivatives were more effective than salicylamide and ulcerogenic activity was variably lowered. PMID- 11440073 TI - Synthesis of new salicylamide derivatives with evaluation of their antiinflammatory, analgesic and antipyretic activities. AB - A new series of pyridazine, pyrazoles, pyrazolidine-3,5-dione, Semicarbazide, thiosemicarbazides, hydantoin, thiohydantoins, 1,2,4-triazoles, S-triazolo[3,4-b] 1,3,4-thiadiazoles incorporated indirectly into salicylamide moiety at position 2 were synthesized. Also the synthesis of novel series of 3-salicylamido-2 hydroxypropyl-amine derivatives were prepared. Several of these compounds were screened for antiinflammatory, analgesic, antipyretic and ulcerogenic activities. PMID- 11440074 TI - Naphthazarin derivatives (VII): antitumor action against ICR mice bearing ascitic S-180 cells. AB - Various analogues of 5,8-dimethoxy-1,4-naphthoquinone (DMNQ) such as 2- or 6-(1 hydroxyiminoalkyl)-DMNQs were prepared and evaluated for the antitumor action. (1 Hydroxyiminoalkyl)-DMNQ derivatives expressed greater antitumor action than (1 hydroxyalkyl)- or acyl-DMNQ derivatives. Moreover, 6-(1-hydroxyiminoalkyl)-DMNQ derivatives expressed higher antitumor action than 2-sudstituted ones, suggestive of a steric effect. Some of 6-(1-propyloxyalkyl)-DMNQ derivatives with an alkyl group of butyl to octyl moiety showed T/C values of >400%. PMID- 11440075 TI - Phytochemical constituents of Artemisia japonica ssp. littoricola. AB - The phytochemical study of the aerial parts of Artemisia japonica ssp. littoricola (Asteraceae) led to the isolation of two acetylenic compounds, (3R) dehydrofalcarinol (2) and (3R)-dehydrofalcarindiol (6), two sesquiterpenes, 1beta, 6alpha-dihydroxy-4(15)-eudesmene (5) and oplodiol (8), and four phenolic compounds, eugenol (1), vanillin (3), 3'-methoxy-4'-hydroxy-trans-cinnamaldehyde (4) and p-hydroxyacetophenone (7). Their structures were determined by chemical and spectroscopic methods. PMID- 11440076 TI - Lignan and neolignan glycosides from Ulmus davidiana var. japonica. AB - Four lignan xylosides and two neolignan glycosides were isolated from the stem and root barks of Ulmus davidiana var. japonica. Their structures were identified as lyoniside, nudiposide, 5'-methoxyisolariciresinol-9'-O-beta-D-xylopyranoside, isolariciresinol-9'-O-1-D-xylopyranoside, rel-trans-dihydrodehydroconiferyl alcohol 4'-O-alpha-L-rhamnopyranoside and icariside E3 by comparison of their spectral data with those reported in the literatures, respectively. PMID- 11440077 TI - Inhibition of calmodulin-dependent calcium-ATPase and phosphodiesterase by various cyclopeptides and peptide alkaloids from the Zizyphus species. AB - The effects of various sedative cyclopeptides and peptide alkaloids from the Zizyphus species on calmodulin-dependent Ca2+-ATPase and phosphodiesterase were investigated. Calmodulin-induced activation of Ca2+-ATPase was strongly inhibited by sanjoinine-A dialdehyde (IC0O, 2.3 microM), -Ah1 (IC50, 4.0 microM), -A (IC50, 4.6 microM), and -G2 (IC50, 7.2 microM), while calmodulin-induced activation of phosphodiesterase was strongly inhibited by both deachuine-S10 (IC30, 4.9 microM) and sanjoinine-D (IC50, 9.0 microM). The inhibitory activity of the various cyclopeptides and peptide alkaloids on Ca2+-ATPase was found to correlate well with their sedative activity. PMID- 11440078 TI - High performance liquid chromatographic analysis of a new proton pump inhibitor KR60436 and its active metabolite O-demethyl-KR60436 in rat plasma samples using column-switching. AB - A fully automated high performance liquid chromatography with column-switching was developed for the simultaneous determination of KR60436, a new reversible proton pump inhibitor, and its active metabolite O-demethyl-KR60436 from rat plasma samples. Plasma sample (50 microl) was directly introduced onto a Capcell Pak MF Ph-1 column (10 x 4 mm I.D.) where primary separation was occurred to remove proteins and concentrate target substances using acetonitrile-potassium phosphate (pH 7, 0.1 M) (2:8, v/v). The drug molecules eluted from MF Ph-1 column were focused in an intermediate column (10 x 2 mm I.D.) by the valve switching step. The substances enriched in intermediate column were eluted and separated on a Vydac 218MR53 column (250 x 3.2 mm I.D.) using acetonitrile-potassium phosphate (pH 7, 0.02 M) (47:53, v/v) at a flow rate of 0.5 ml/min when the valve status was switched back to A position. The method showed excellent sensitivity (detection limit of 2 ng/ml) with small volume of samples (50 microl), good precision and accuracy, and speed (total analysis time 24 min) without any loss in chromatographic efficiency. The response was linear (r2 > or = 0.999) over the concentration range of 5-500 ng/ml. PMID- 11440079 TI - Cytotoxic constituents of Psoralea corylifolia. AB - A coumestan derivative, psoralidin (1) was found to be a cytotoxic principle of the seeds of Psoralea corylifolia L. (Leguminosae) with the IC50 values of 0.3 and 0.4 microg/ml against the HT-29 (colon) and MCF-7 (breast) human cancer cell lines, respectively. A coumarin, angelicin (2) was also isolated as a marginally cytotoxic agent along with an inactive compound, psoralen (3) from the plant. The isolates 1-3 were not active against the A541 (lung) and HepG2 (liver hepatoma) cancer cell lines. PMID- 11440080 TI - Comparisons between white ginseng radix and rootlet for antidiabetic activity and mechanism in KKAy mice. AB - The mechanisms responsible for the antidiabetic activity of both the white ginseng radix (Ginseng Radix Alba, GRA) and the rootlet (Ginseng Radix Palva, GRP) were investigated. After a four week oral administration, the fasting blood glucose levels in the GRA- and GRP-treated groups were lower when compared to the control group. To elucidate the hypoglycemic mechanism(s) of the ginseng radices, glucose absorption from the small intestine, hepatic hexokinase and glucose-6 phosphatase activities, in addition to PPAR-gamma expression in adipose tissue were examined. The results strongly suggest that GRA can improve hyperglycemia in KKAy mice, possibly by blocking intestinal glucose absorption and inhibiting hepatic glucose-6-phosphatase, and GRP through the upregulation of adipocytic PPAR-y protein expression as well as inhibiting intestinal glucose absorption. PMID- 11440081 TI - Nasal absorption of procyclidine in rats and dogs. AB - Nasal absorption of procyclidine, a synthetic anticholinergic compound, was investigated in Wistar rats and Beagle dogs. The dosing solution was prepared by dissolving 14C-procyclidine in 50% ethanolic saline. The dosing solution was administered intravenously and intranasally to rats at a dose of 0.6 mg/kg (i.e., 60 microl/kg in the form of a 1% w/v solution), and intravenously, orally and intranasally to dogs at a dose of 0.3 mg/kg (i.e., 6 microl/kg in the form of a 5% w/v solution). Blood samples were taken from an artery of the animals through the catheter for periods of 1200 (for rats) and 1,440 min (for dogs), and the radioactivity in the samples was determined by liquid scintillation counting. The nasal bioavailability of procyclidine in rats and dogs, based on the radioactivity, was calculated to be 81.1 and 98.6%, respectively. In both rats and dogs, the plasma profiles of procyclidine following nasal administration were very close to those following intravenous administration, leading to nearly superimposable profiles between the two protocols. In dogs, nasal administration resulted in significantly higher plasma concentrations during the first 30 min period compared to oral administration, suggesting the superiority of the nasal route over the oral route in terms of a prompt expression of the pharmacological effect of the drug. The results obtained in this study indicate that procyclidine is rapidly and nearly completely absorbed via the nasal route. In conclusion, nasal administration represents a viable alternative to intravenous administration in the case of procyclidine. PMID- 11440082 TI - Enhanced skin permeation of a new capsaicin derivative (DA-5018) from a binary vehicle system composed of isopropyl myristate and ethoxydiglycol. AB - DA-5018, a recently synthesized capsaicin analog, appears to possess potent analgesic activity when administered topically. The objective of this study is to test the feasibility of the topical administration of this compound. Specifically, our goal was to identify vehicle system that permit a reasonable transdermal permeation of the compound in mice. Among the vehicles examined, isopropyl myristate (IPM) showed the largest in vitro permeability across the intact skin (83.6 +/- 5.42 microl/cm2/h). However, due to the limited solubility of DA-5018 in IPM (0.53 mg/ml), the maximal flux from the IPM medium remained at only 44.3 +/- 2.87 microg/cm2/hr. In order to increase the flux, addition of better solvents for DA-5018 was attempted, under the assumption that flux is the result of both solubility and permeability. Ethoxydiglycol (EG) and oleic acid (OA) were selected as examples of good solvents. The addition of EG or OA to IPM at a 1:1 volume ratio resulted in a comparable increase in the solubility of the compound (i.e., to 61.1 and 50.2 mg/ml for EG and OA, respectively). However, the addition of EG at a 1:1 volume ratio, for example, increased the flux 6.3 fold (i.e., 279 microg/cm2/hr), while OA, at a 1:1 volume ratio, decreased the flux 5 fold (i.e., 9.26 microg/cm2/hr). The mechanism of this discrepancy between EG and OA was investigated by measuring the permeabilty of DA-5018 across the stratum corneum-removed skin of the mouse, under the hypothesis that the viable skin layer may serve as a barrier for the permeation of lipophilic substances such as DA 5018. The permeability of DA-5018, from the medium of EG or OA, across the viable skin differed greatly for EG (0.41 microl/cm2/hr) and OA (0.086 microl/cm2/hr), suggesting that a higher permeability across the viable skin layer is needed for the second solvents. The maximum flux across the intact skin was achieved for DA-5018 when EG was added to IPM at a 1:1 volume ratio. Thus, the use of a binary system appears to be the bes approach for realizing the transdermal delivery of DA-5018 at a reasonable rate. PMID- 11440083 TI - Radiation technology in the preparation of polyethylene oxide hydrophilic gels and immobilization of proteases for use in medical practice. AB - This paper deals with the development of a technology for making a hydrophilic gel of polyethylene oxide reception in which radiating ability is employed to cause cross-linking of polymers in a water solution. The gel of polyethylene oxide was shown to be non-toxic, contain 5-50% of polymer and be useful in composite medicinal forms along with biologically active substances including Bac. subtilis proteases. Proteases immobilized in the gel possess high thermal stability and proteolytic activity and are readily applied in medicine. The effect of immobilized proteolytic and glucolytic enzymes of Bac. subtillis (Immozimase) on the warm ischemia-reperfusion (I/R) which can cause hepatic and jejunum injury was also studied. These enzymes were immobilized on water-soluble polymer polyethylene glycol by means of an electron beam. The number of degranulated mast cells as well as serum ALT after I/R in the group with Immozimase was decreased to almost half as compared with the control group. Pretreatment with Immozimase resulted in significant reduction of hepatic and gut neutrophil accumulation as compared with control animals. It was concluded that Immozimase has a protective effect for hepatic and gut ischemia/reperfusion, and this effect seems to be associated with prevention of leukocyte accumulation. PMID- 11440084 TI - Impairments of learning and memory following intracerebroventricular administration of AF64A in rats. AB - Three types of learning and memory tests (Morris water maze, active and passive avoidance) were performed in rats following intracerebroventricular infusion of ethylcholine aziridium (AF64A). In Morris water maze, AF64A-treated rats showed the delayed latencies to find the platform from 6th day after the infusion. In pretrained rats, AF64A caused the significant delay of latency at 7th day, but not 8th day. In the active avoidance for the pre-trained rats, the escape latency was significantly delayed in AF64A-treatment. The percentages of avoidance in AF64A-treated rats were less increased than those in the control. Especially, the percentage of no response in the AF64A-treated rats was markedly increased in the first half trials. In the passive avoidance, AF64A-treated rats shortened the latency 1.5 h after the electronic shock, but not 24 h. AF64A also caused the pretrained rats to shorten the latency 7th day after the infusion, but not 8th day. These results indicate that AF64A might impair the learning and memory. However, these results indicate that the disturbed memory by AF64A might rapidly recover after the first retrain. Furthermore, these results suggest that AF64A may be a useful agent for the animal model of learning for spatial cognition. PMID- 11440085 TI - Influence of quinine on catecholamine release evoked by cholinergic stimulation and membrane depolarization from the rat adrenal gland. AB - The present study was attempted to investigate the effect of quinine on secretion of catecholamines (CA) evoked by cholinergic stimulation and membrane depolarization from the isolated perfused rat adrenal gland. The perfusion of quinine (15-150 microM) into an adrenal vein for 60 min produced dose- and time dependent inhibition in CA secretion evoked by ACh (5.32 x 10(-3) M), high K+ (5.6 x 10(-2) M), DMPP (10(-4) M for 2 min), McN-A-343 (10(-4) M for 2 min), cyclopiazonic acid (10(-5) M for 4 min) and Bay-K-8644 (10(-5) M for 4 min). Also, under the presence of pinacidil (10(-4) M), which is also known to be a selective potassium channel activator, CA secretory responses evoked by ACh, high potassium, DMPPF McN-A-343, Bay-K-8644 and cyclopiazonic acid were also greatly reduced. When preloaded along with quinine (5 x 10(-5) M) and glibenclamide (10( 6) M), a specific blocker of ATP-regulated potassium channels, CA secretory responses evoked by ACh, high potassium, DMPP, McN-A-343, Bay-K-8644 and cyclopiazonic acid were recovered as compared to those of quinine-treatment only. Taken together, these results demonstrate that quinine inhibits CA secretion evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors as well as by membrane depolarization through inhibiting influx of extracellular calcium and release in intracellular calcium in the rat adrenomedullary chromaffin cells. These findings suggest that activation of potassium channels may be involved at least in inhibitory action of quinine on CA secretion from the rat adrenal gland. PMID- 11440086 TI - Antiallergic action of Magnolia officinalis on immediate hypersensitivity reaction. AB - We studied the effect of aqueous extract of Magnolia officinalis bark (Magnoliaceae) (MOAE) on the immediate hypersensitivity reaction. MOAE (0.01 to 1 g/kg) dose-dependently inhibited compound 48/80 induced systemic anaphylaxis in rats. MOAE (0.1 and 1 g/kg) also significantly inhibited local immunoglobulin E (IgE)-mediated passive cutaneous anaphylactic reaction. When MOAE was pretreated at concentrations ranging from 0.01 to 1 g/kg, the levels of plasma histamine were reduced in a dose-dependent manner. MOAE (0.001 to 1 mg/ml) dose-dependently inhibited the histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-dinitrophenyl (DNP) IgE. The level of cyclic AMP (cAMP) in RPMC, when MOAE was added, significantly increased compared with that of the normal control. Moreover, MOAE (0.01 to 1 mg/ml) had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-alpha production from RPMC. These results indicate that MOAE inhibits immediate hypersensitivity reaction in vivo and in vitro. PMID- 11440087 TI - Allergenicity test of genetically modified soybean in Sprague Dawley rats. AB - Allergenicity of genetically-modified (GM) soybean was evaluated in male Sprague Dawley rats. To confirm the GM soybean used in this study, the polymerase chain reaction (PCR) was performed using the chromosomal DNA of soybeans. The PCR result provided the clear discrimination of genetically-modified (GM) soybeans. To evaluate the allergenicity of GM soybean and non-GM control one, the soybean homogenate was sensitized subcutaneously 3 times a week for 3 weeks. The doses of soybean were 0, 2 and 20 mg/kg in the protein basis. A week after the last sensitization, antisera were recovered from individual animals. When the sera were injected intradermally on the clipped back of unsensitized rats with various dilutions, followed by a challenge with 20 mg/kg of soybean homogenate containing 1% Evans blue, no sign of passive cutaneous anaphylaxis reaction was detected. In addition, when the sera were treated in the cultures of peritoneal mast cells, the increase of histamine release by anti-(GM soybean) sera was not observed when compared to that by anti-(non-GM soybean) sera. The present results indicate that the GM soybean might not act as a strong allergen in male Sprague Dawley rats. PMID- 11440088 TI - Recurrent abdominal pain. A world-wide problem of organic, functional and psychosomatic aetiology. AB - This spring, Acta Paediatrica is publishing five original articles underlining the importance of recurrent abdominal pain (RAP) and illustrating the scientific and diagnostic difficulties involved in understanding it. Our knowledge of organic causes is increasing with improved instruments for investigation, including 24-h pH monitoring and endoscopy, but the difficulty in how to link findings of organic abnormalities conclusively to the symptom RAP has to be further elaborated and discussed. There is an old belief that many cases of RAP have a psychogenic aetiology. Psychometric tests in past decades, however, have not supported this belief, but if the reaction primarily is one of negative stress, than we should be looking for negative stress reactions, not for psychopathology. RAP of psychosomatic origin is not just a matter of excluding organic disease, but a diagnosis that must be built on firm positive diagnostic criteria, criteria that have not yet been elaborated for a broader public. PMID- 11440089 TI - Which place for bismuth subsalicylate in the treatment of enteric infections? AB - ORS has led to improved outcome of acute gastroenteritis in both industrialised and developing countries. In both settings there is an increasing demand for active therapy to reduce the duration of diarrhoea and its complications. Persistent diarrhoea is a major consequence of intestinal infections and is responsible for a high number of deaths in poor countries. Bismuth subsalicylate has been used for treatment of acute diarrhoea, with preliminary promising results. In this issue of Acta Paediatrica, a trial with BSS is essential. However the results were marginal and did not justify a mass scale use of BSS, also because of poor cost efficacy rate. PMID- 11440090 TI - The efficacy of bismuth subsalicylate in the treatment of acute diarrhoea and the prevention of persistent diarrhoea. AB - A controlled, randomized, double-blind study in Bangladeshi children (ages 4-36 mo) with acute diarrhoea was undertaken to determine whether bismuth subsalicylate (BSS) would prevent the development of persistent diarrhoea (PD) in young children. The children were randomized to two groups: 226 were given liquid oral BSS, (as Pepto-Bismol), 100 mg/kg/d for 5 d; 225 were given placebo of identical appearance. On admission to the study, the two groups were comparable both clinically and microbiologically. Rotavirus was found in 56% of all the children, and enterotoxigenic E. coli in 31% of a subsample studied. Children treated with BSS had less severe and less prolonged illness than those treated with placebo (p = 0.057). There was, however, no difference in the development of PD between the two groups (8% and 11%). Unexpectedly, patients treated with BSS gained significantly more weight (2.3%) than those treated with placebo (0.5%; p < 0.001) during the course of the study. No toxicity of BSS was detected. CONCLUSION: Treatment with BSS had a modest therapeutic effect on acute diarrhoea, as has been previously demonstrated, but with no suggestion of a therapeutic effect on the prevention of persistent diarrhoea in this group of patients. PMID- 11440091 TI - Prediction of bacterial meningitis in children with meningeal signs: reduction of lumbar punctures. AB - Physicians often have to perform a lumbar puncture to ascertain the diagnosis in patients with meningeal signs, because of the serious consequences of missing bacterial meningitis. The aim of this study was to derive and validate a clinical rule to predict bacterial meningitis in children with meningeal signs, to guide decisions on the performance of lumbar punctures. Information was collected from records of patients (aged 1 mo to 15 y) consulting the emergency department of the Sophia Children's Hospital between 1988 and 1998 with meningeal signs. Bacterial meningitis was defined as cerebrospinal fluid (CSF) leucocyte count >5 cells microl(-1) with a positive bacterial culture of CSF or blood. The diagnostic value of predictors was judged using multivariate logistic modelling and area under the receiver operating characteristic curves (ROC area). In the derivation set (286 patients, years 1988-1995) the duration of the main complaint, vomiting, meningeal irritation, cyanosis, petechiae and disturbed consciousness were independent clinical predictors of bacterial meningitis. The ROC area of this model was 0.92. The only independent predictor from subsequent laboratory tests was the serum C-reactive protein concentration, increasing the ROC area to 0.95. Without missing a single case, this final model identified 99 patients (35%) without bacterial meningitis. Validation on 74 consecutive patients in 3 subsequent years (1996-1998) yielded similar results. CONCLUSION: This prediction rule identifies about 35% of the patients with meningeal signs in whom a lumbar puncture can be withheld without missing a single case of bacterial meningitis. For the individual patient this prediction rule is valuable in deciding whether or not to perform a lumbar puncture. PMID- 11440092 TI - High prevalence of stress hyperglycaemia in children with febrile seizures and traumatic injuries. AB - Although hyperglycaemia is relatively frequent in the course of severe illnesses and may be looked upon as the possible result of an uncoordinated insulin response to the increased glucose that the body may need during periods of stress, it is generally agreed that it does not constitute a prediabetic condition. Numerous studies have aimed to explain the pathophysiology of this occurrence but none has looked at which conditions are more prone to develop stress hyperglycaemia (SH). Therefore, the aim of this study was to evaluate the main clinical conditions that may be associated with SH in children. A total of 1199 children was studied: 833 children (439 M, 394 F, mean age 5.2 +/- 4.5 y) admitted for an acute illness or injury constituted the stress-exposed group, while 366 children (222 M, 144 F, mean age 6.2 +/- 4.6 y) admitted for elective minor surgery represented the stress-unexposed group and were considered as the control group. SH was defined as plasma glucose concentrations > or = 8.3 mmol l( 1) during an acute illness. Stress-exposed patients had significantly higher glycaemic levels than controls (5.6 +/- 1.4 vs 4.7 +/- 0.7 mmol l(-1); p < 0.0001). SH was found in 41 (4.9%) stress-exposed patients and in none of the controls. SH was significantly more prevalent in children affected by febrile seizures (12.9%) or traumatic injuries (11.7%; p < 0.008 and p < 0.02, respectively, vs other diagnoses). A significant correlation was found between glycaemia and systolic pressure (r = 0.1; p < 0.01), white cell count (r = 0.12; p < 0.0003) and body temperature (r = 0.16; p < 0.0001). SH was more frequent in patients with body temperature > 39 degrees C (14%) than in those with a temperature < or = 39 degrees C (4%; p < 0.0008). SH was more prevalent in clinical conditions of fever associated with seizures or pain (12.9% and 12.5%, respectively) than fever alone (4.4%). After a mean period of 3.5 +/- 0.6 y of follow-up none of the hyperglycaemic patients had developed diabetes mellitus. CONCLUSION: Traumatic injuries, febrile seizures or conditions in which an elevated body temperature may be found are frequently associated with SH in children. In the presence of these conditions specific studies directed towards unmasking a prediabetic state may be unnecessary. PMID- 11440093 TI - Effect of intravenous immunoglobulin on inhibiting peripheral blood lymphocyte apoptosis in acute Kawasaki disease. AB - This study aimed to explore the therapeutic mechanism of intravenous immunoglobulin (IVIG) for Kawasaki disease (KD). Peripheral blood lymphocytes (PBLs) obtained from 26 children with KD and 20 age-matched healthy children were stimulated with anti-CD3 monoclonal antibody (mAb), and the percentage of apoptotic cells and DNA fragmentation were assayed at 0, 12, 24, 48 and 72 h in vitro. The patients were divided into two groups: one treated with aspirin combined with IVIG (n = 16) and one treated with aspirin alone (n = 10). PBLs were stimulated by phytohaemagglutinin to evaluate the lymphocyte proliferative response. Compared with normal controls, the apoptotic cell percentage and the DNA fragmentation were markedly decreased (p < 0.001) and delayed in PBLs from KD patients. After IVIG treatment, the decreased percentage of apoptotic cell and delayed DNA fragmentation were restored to the state of the normal controls, accompanied by a fast clinical remission compared with the aspirin-alone group. The lymphocyte proliferative response was also decreased 3-5 d after IVIG therapy (p < 0.001). CONCLUSION: The results suggest that decreased PBL apoptosis may be involved in the pathogenesis of KD. The therapeutic mechanism of IVIG in KD may be partially due to the reversal of the inhibited lymphocyte apoptosis, and may have implications for other autoimmune diseases with inefficient lymphocyte apoptosis. PMID- 11440094 TI - Association between urinary tract infection, renal damage and birth size. AB - The aim of this study was to investigate whether birth size is associated with permanent renal damage in children with urinary tract infection (UTI). A cohort of 1221 children under 16 y of age was diagnosed with their first symptomatic UTI between 1970 and 1979. Of these, 74 had urographic renal scarring in childhood and 57 were re-examined as adults. The birth files of 48 of these patients (35F, 13M) were available, and birthweight and birthlength in relation to gestational age were analysed and compared with a Swedish reference population. Children who had renal damage without vesicoureteric reflux were significantly smaller at birth (median weight, -0.76 SDS) compared with both children who had renal damage and reflux (median weight, -0.01 SDS) and the reference population. CONCLUSION: The demonstration of low birthweight among children with UTI and renal damage but no reflux suggests that low birthweight may be a risk factor for the development of renal damage. PMID- 11440095 TI - Personality profiles and heart rate variability (vagal tone) in children with recurrent abdominal pain. AB - The aim of the study was to explore psychological factors and autonomic activity in children with recurrent abdominal pain and to compare them with those in a control group of healthy children. The Personality Inventory for Children was used for assessment of developmental, emotional and psychosocial factors in 25 children with recurrent abdominal pain (age, 7-15 y). Parasympathetic and sympathetic functions in these children and in 23 healthy control subjects (age, 7-13 y) were also investigated, non-invasively using a computerized polygraph. Vagal tone (parasympathetic function) was indexed by calculation of respiratory sinus arrhythmia in beats/min. Skin conductance (sympathetic function) was recorded by the constant current method. On the Personality Inventory for Children, 16 patients had high scores on somatic concern. Several patients had scores in the clinical range for depression, withdrawal and anxiety, but the mean scores for these personality profile scales were well within the normal range of healthy children. Interestingly, there was a spike on the L (Lie)-scale for most of the patients and 15 patients had scores above or close to the clinical cut-off value. As compared with the scores in healthy children, vagal tone and sympathetic tone were normal. CONCLUSION: Many children with recurrent abdominal pain have scores in the clinical range for depression, withdrawal, anxiety and L scale indicating coping problems, denial and a trend towards somatic concern that may contribute to the evolution of abdominal pain. Autonomic nerve activity was not disturbed in these children. PMID- 11440096 TI - Organic abnormalities in recurrent abdominal pain in children. AB - Using an investigation protocol, the aim of this study was to determine the frequency of organic abnormalities in children with recurrent abdominal pain, as new diagnostic approaches may reveal a higher prevalence of organic disease in this group than has been found in most studies. Included in the study were 44 children (mean age 8.3 y; 2-15) with more than three bouts of abdominal pain severe enough to affect the daily activities of the child and lasting more than 3 mo. The investigation covered a detailed medical story, a physical examination, blood, urine and stool samples. The somatic investigation was completed by abdominal X-ray and ultrasound, lactose-breath-hydrogen test and 24-h pH monitoring in the lower oesophagus. A Child Behaviour Checklist was completed to assess psychosocial aspects of the illness. The blood, urine and stool samples were normal, and abdominal ultrasound did not give any results related to the symptoms. Constipation was diagnosed in 7 patients (16%); 9 patients (21%) had gastro-oesophageal reflux and oesophagitis was found in another 3 children. One child had nodular antral gastritis with colonization by Helicobacter pylori, and three children had pathological lactose-breath-hydrogen tests. Twenty-four children (55%) did not have any signs of organic disease. The total score for the CBCL was in the normal range in 89%. CONCLUSION: Our observations indicate a higher proportion of organic abnormalities in recurrent abdominal pain than has been found in most previously reported studies, though a multicausal approach seems important. PMID- 11440097 TI - Recurrent abdominal and flank pain in children with idiopathic hypercalciuria. AB - OBJECTIVE: To evaluate the role of idiopathic hypercalciuria (IH) as a cause of recurrent abdominal pain (RAP) in children. PATIENTS AND METHODS: We retrospectively reviewed the medical records of 124 children referred for various complaints who had 24-h urine calcium excretion greater than 2 mg/kg/d or random urine calcium-creatinine ratio greater than 0.18 mg/mg. RESULTS: Fifty-two children with various clinical complaints had RAP or flank pain. These comprised of 22 males and 30 females, 9 mo to 15.9 y of age, mean 6.7 +/- 3.5 y. A family history of urolithiasis was present in 50% of all the children. Only 6 of the 52 children with abdominal pain had renal stones. In addition to abdominal pain, 27 children had hematuria and 10 had urinary incontinence. Mild metabolic acidosis was present in three children, parathyroid hormone activity elevated in two and serum vitamin D activity was increased in nine. All children were treated with increased fluid intake and a reduction in dietary sodium and oxalate and some required treatment with thiazide and antispasmodics. Forty-five cases responded to treatment, 5 failed to improve from therapy, and 2, which were not followed up as patients, were not available. CONCLUSION: We describe 52 children with RAP or back pain due to IH and recommend that IH be considered in the differential diagnosis of RAP in childhood. PMID- 11440098 TI - Do normal leucocyte count and C-reactive protein value exclude acute appendicitis in children? AB - The present study aimed to determine the role of leucocyte count and C-reactive protein (CRP) measurements in the diagnosis of acute appendicitis in children. In particular, children with acute appendicitis but normal leucocyte count and CRP level were sought. The present study protocol was identical to those used in earlier studies on adult patients with suspected acute appendicitis. The mean preoperative leucocyte count and CRP value in 100 consecutive children with an uninflamed appendix at appendicectomy (group A) and in 100 consecutive patients with acute appendicitis (group B) were calculated. The numbers of patients with (i) both values normal, (ii) only leucocyte count raised, (iii) only CRP level raised, and (iv) both values raised were calculated in both groups A and B. Leucocyte count effectively (p < 0.001) separated children with uninflamed appendix (mean +/- SEM 10.2 +/- 0.4 x 10(9) l(-1)) from those with acute appendicitis (15.0 +/- 0.4 x 10(9) l(-1)), but the CRP value was of no use in this respect (p = 0.866; 31 +/- 4 mg l(-1) and 30 +/- 4 mg l(-1)). The most conspicuous finding was that in children with acute appendicitis, both values were normal in 7 out of 100 patients. CONCLUSION: Contrary to adult patients, normal leucocyte count and CRP value do not effectively exclude acute appendicitis in children. PMID- 11440099 TI - Cholesterol dynamics in the foetal and neonatal brain as reflected by circulatory levels of 24S-hydroxycholesterol. AB - Oxysterols, particularly those hydroxylated in the steroid side-chain, are formed from cholesterol by specific cytochrome P450 enzymes and may facilitate elimination of cholesterol from extrahepatic sources. In humans, the greatest portion of circulating 24S-hydroxycholesterol (24S-OH-Chol) is derived from the brain and the absolute concentration depends on age. In the present study, concentrations of 24S-OH-Chol and for comparison 27-OH-Chol were determined by a highly sensitive isotope dilution method using gas chromatography-mass spectrometry in serum samples from normal preterm and term neonates and those with Rhesus haemolytic disease, taken serially for diagnostic purposes. Serum concentrations of cholesterol, 24S-OH-Chol and 27-OH-Chol were similar in venous versus arterial cord blood of 6 term neonates. Serum concentrations of 24S-OH Chol and 27-OH-Chol in 12 small for gestational age (SGA) preterm neonates were significantly lower than those in 12 appropriate for gestational age (AGA) preterm neonates (p < 0.001), and also lower than those in 12 SGA (0 < 0.001) and 12 AGA term neonates (p < 0.05). Serum cholesterol was significantly higher in preterm than in term neonates (p < 0.001). 24S-OH-Chol serially determined in 8 infants with Rhesus haemolytic disease increased 5-6-fold during the first 3 mo after birth (from 42 +/- 20 ng ml(-1) to 227 +/- 71 ng ml(-1)). 27-OH-Chol increased simultaneously from 30 +/- 14 ng ml(-1) to 100 +/- 39 ng ml(-1). CONCLUSION: Serum concentrations of 24S-OH-Chol increased 5-6-fold after birth. This could be an indication of normal cholesterol metabolism in the developing neonatal brain. PMID- 11440100 TI - Early oral behaviour in preterm infants during breastfeeding: an electromyographic study. AB - The objectives were to increase the understanding of the characteristics of oral behaviour during breastfeeding in preterm infants and to validate direct observations of infant sucking. Twenty-six infants were investigated once by simultaneous observation and surface electromyography (EMG) at 32.1-37.1 postmenstrual weeks. The orbicularis oris muscle was used for data analysis, as it provided the most distinct registrations. High correlation coefficients were observed with respect to classifications of EMG data by two raters on the number of sucks per burst (r = 0.97) and duration of sucking bursts (r = 0.99). The agreement between direct observations of sucking and EMG data was high. The median for mean number of sucks per burst was 8 (range 2-33) and for longest burst 28 (5-96) sucks. Sucks with low and very high intensity constituted a median of 14 (1-94)% and 25 (0-87)% of all sucks. The range in mean sucking frequency was 1.0-1.8 sucks s(-1). Suck duration ranged from 0.6 to 1.1 s. There was a considerable variation between infants in the extent of mouthing. No association with maturational level appeared for any of the components in oral behaviour. CONCLUSION: EMG data provided evidence of early sucking competence in preterm infants during breastfeeding, with wide individual variations. Surface EMG and direct observation are recommended as valid methods in the evaluation of breastfeeding behaviour in preterm infants. PMID- 11440101 TI - Shorter PFA-100 closure times in neonates than in adults: role of red cells, white cells, platelets and von Willebrand factor. AB - In neonates, despite poor platelet function in various in vitro tests, closure times (CTs) in PFA-100 measurements are shorter than in adults. Neonates have a higher polymeric von Willebrand factor (vWF). They also have a higher haematocrit and higher white blood cell count than adults. which may interfere with the evaluation of platelet and vWF function by means of the PFA-100 in neonates. To assess the role of different blood constituents on neonatal CTs, red blood cell, platelet and white blood cell counts in cord blood were modified. These modifications did not provide any evidence that the difference in number between adult and neonatal blood cells was responsible for shorter neonatal CTs. In further experiments, platelets and/or vWF were inhibited by means of abciximab and anti-vWF antibody, and mixing experiments with neonatal platelet-rich and platelet-poor plasma were performed. The results showed that short cord blood PFA 100 CTs were caused by a constituent of neonatal platelet-poor plasma, probably the neonatal high multimeric vWF. CONCLUSION: This study demonstrates that CTs in neonates are dependent on the same components, platelets and vWF, as in adults, making it likely that the PFA-100 can be used in neonates in the same way as in adults to investigate platelet and vWF function. PMID- 11440102 TI - Prevalence of overweight and obesity in children and adolescents in a county in Sweden. AB - Overweight among young people in Sweden is increasing. The aim of the present study was to investigate the frequency of overweight and obesity based on body mass index (BMI) values among children and adolescents. Overweight was defined as a BMI value > or = 91st percentile and obesity as a BMI value > 98th percentile on an international reference BMI curve. The study population included boys and girls from four age groups: 9, 12, 15 and 18 y. The data consisted of self reported measures of height and weight that were obtained from questionnaires used in a cross-sectional study in December 1997. A validation study was performed using a part of the study population. A total of 7011 (81.7%) participants completed the questionnaire. The correlation between self-reported estimations and objective measures of height and weight was high in the oldest age groups (0.88-0.98), but lower in the 9-y-old age groups (0.37-0.72). These self-reported estimations in the 9-y-olds were excluded from further analysis. It was found that 12.3%, 11.6% and 11.4% of the boys in the 12-, 15- and 18-y-old age groups and 6.8%, 5.5% and 4.8% of the girls in the same age groups were overweight and 7.9%, 8.9% and 7.3% of the boys and 5.1%, 4.2% and 3.9% of the girls were obese. CONCLUSION: The prevalence of overweight and obesity was found to be high in the study population and is a serious public health problem. The prevalence of obesity was significantly higher (p < 0.05) in 15-y-old boys living in rural areas than in city and town dwellers of the same age. PMID- 11440103 TI - Changes in body weight and body mass index (BMI) in teenage girls prior to the onset and diagnosis of an eating disorder. AB - Growth charts from the school health services were used to study weight changes prior to the onset and diagnosis of eating disorders (ED) with weight loss in 122 teenage girls. In these girls menarche started at 12.6 +/- 1.0 y (mean +/- SD), which is the same for the general population. At their maximal premorbid weight, the girls were 14.6 +/- 1.2 y of age and had a higher weight and body mass index (BMI) than the general population, as evidenced by standard deviation scores (SDS) above zero for weight (0.65 +/- 0.95, p < 0.001) and BMI (0.61 +/- 0.94, p < 0.001). Weight gain prior to the onset of weight loss followed two different patterns. Eighty-three (68%) girls tracked upwards through the weight curves to reach their maximal weight, thus putting on weight at a higher rate than expected. Thirty-nine (32%) girls tracked downwards through the weight curves to reach their maximal weight, but did so starting from a weight and BMI at well above average. CONCLUSIONS: Weight gain in teenage girls prior to the onset of an ED deviates from that of the general population. Being or becoming heavier and less lean than their peers could influence body image and decisions on dieting, which, in turn, may be the start of the development of an ED in the otherwise predisposed. The deviation from normal tracking patterns prior to the onset of disease also causes problems in diagnostic procedures and in setting treatment targets for weight gain, since "normal" or expected weight becomes more difficult to establish. PMID- 11440104 TI - Causes of childhood deaths in Bangladesh: an update. AB - Knowledge of the causes of child death is important for health-sector planning since they relate to available interventions. Little is known about causes of child death in Bangladesh from the conventional sources since there is no vital registration system and very few deaths are attended by a qualified physician. To determine the cause structure of child deaths, verbal autopsy interviews were conducted in the Bangladesh Demographic and Health Survey (BDHS) 1993/94 national sample. Verbal autopsy is a method of finding out the causes of death based on an interview with the next of kin or other caregivers. Between BDHS 1993/94 and BDHS 1996/97, 1-4-y-old child mortality in Bangladesh declined by about 27.0%. This impressive decline prompted a verbal autopsy study using the BDHS 1996/97 national sample to determine whether the cause structure had changed. The same verbal autopsy instrument and methods to collect the data and the same computer algorithm to assign causes of death were used in both surveys. Comparison of BDHS 1993/94 and 1996/97 cause-specific mortality rates revealed that deaths due to almost all causes had declined, although significantly so only for acute respiratory infections (ARI), persistent diarrhoea and drowning. Deaths due to neonatal tetanus, acute watery diarrhoea and undernutrition had not decreased at all. CONCLUSION: Despite an impressive decline in deaths due to ARI, this condition remains the most important known cause of death in Bangladeshi children. Neonatal tetanus and measles together account for about 10% of deaths in children under 5 y. Further improvements in child survival are possible by improving access to and quality of available child survival interventions. PMID- 11440105 TI - Lower limb and back pain in Guillain-Barre syndrome and associated contrast enhancement in MRI of the cauda equina. AB - This study assesses the frequency of lower limb and back pain in children with Guillain-Barre syndrome and reviews the magnetic resonance imaging results of those undergoing spinal imaging. Over an 8-y period, nine children presented with various combinations of severe back pain, leg pains, impairment of gait and bladder dysfunction. Guillain-Barre syndrome was confirmed on clinical examination and peripheral electrophysiology (n = 8). Magnetic resonance imaging in four patients, following contrast injection, showed enhancement of the cauda equine and, additionally, of the cervical nerve roots in one of the patients. A further patient, who was not scanned with contrast, had abnormal thickening of the lumbar roots. Carbamazepine and steroids were effectively used for analgesia in three cases. All the patients recovered. CONCLUSION: Guillain-Barre syndrome should be considered in the differential diagnosis of children presenting with back and/or leg pain. Early diagnosis ensures prompt monitoring for autonomic dysfunction and respiratory compromise. PMID- 11440106 TI - Low-density lipoprotein apheresis in a patient aged 3.5 years. AB - A 3.5 y-old girl carrying a severe mutation of the LDL-receptor gene known as "FH Pavia", affected by homozygous familial hypercholesterolaemia (FH), and at high risk of developing coronary artery atherosclerosis was treated with selective dextran sulphate cellulose (DSC) column low-density lipoprotein apheresis (LDL a). This is the youngest patient ever treated with LDL-a. Plasma total cholesterol (982 mg/dl) and LDL-cholesterol (939 mg/dl) (T-Chol, LDL-Chol) levels at baseline showed a transient decrease: -13.4%, and -16.8%, respectively, after 9 mo of combined treatment with a diet, cholestyramine (max. 12 g/d) and atorvastatin (max. 30 mg/d). However, the drugs were discontinued because of intolerance and an increase in aminotransferases and creatine phosphokinase in the plasma. Moreover, after 9 mo of this therapy, the mean plasma T-Chol and LDL Chol levels were still high (930 mg/dl and 869.5 mg/dl, respectively). Therefore, 9 consecutive treatments with LDL-a were carried out every 15 d (plasma volumes treated: 1000-1700 ml). Mean plasma T-Chol, LDL-Chol, triglycerides (TG), and Lp(a) decreased significantly: -75.5%, -77.2%, -67.5% and -50.8%, respectively. HDL-cholesterol (HDL-Chol) concentration was considerably decreased immediately after apheresis because of haemodilution (X: -45.1%). CONCLUSION: LDL-a treatment improved the plasma apo B 100-containing lipoproteins--LDL, Lp(a)--profile in a homozygote with a severe inherited disorder in which coronary artery atherosclerosis frequently has its clinical onset before 10 y of age. At the time of this report, no significant side effects had been observed. PMID- 11440107 TI - Cornelia de Lange syndrome associated with cecal volvulus: report of a case. AB - Cornelia de Lange syndrome is known to be occasionally associated with gastrointestinal malformation. However, the occurrence of cecal volvulus in such anomalies is very rare. We report a 15-y-old Japanese boy with Cornelia de Lange syndrome associated with a cecal volvulus secondary to non-fixation of the cecum and ascending colon. The patient was admitted to our hospital because of abdominal pain and bilious vomiting. At emergent laparotomy, cecal volvulus with non-fixation of the ascending colon and cecum was diagnosed. Release of the cecal volvulus and the fixation of the cecum and ascending colon to the right retroperitonium were performed. The postoperative course was uneventful, except for sepsis caused by aspiration pneumonia soon after the operation. CONCLUSION: Clinicians treating patients with this syndrome should be aware that such patients may be at risk for developing severe gastrointestinal anomalies, including cecal volvulus. PMID- 11440108 TI - Clostridium tertium in cerebrospinal fluid of a premature neonate with necrotizing enterocolitis: contamination or real? PMID- 11440109 TI - Infantile systemic hyalinosis: a case with atypical prolonged survival. PMID- 11440110 TI - Characterization of binding between the rat small intestinal brush-border membrane and dietary proteins in the sensory mechanism of luminal dietary proteins. AB - Dietary proteins are recognized by the gastrointestinal tract to display physiological functions, however, the sensory mechanism of the intestinal mucosa is not known. We examined binding properties between the rat small intestinal brush-border membrane (BBM) and proteins by using a surface plasmon resonance biosensor. BBM and solubilized BBM prepared from the rat jejunum bound to casein immobilized on the sensor surface, but not to bovine serum albumin. The ileal BBM showed less binding to casein than the jejunal BBM. Solubilized BBM binding to immobilized alpha-casein was slightly inhibited by aminopeptidase inhibitors, but still more inhibited by addition of casein with the inhibitors. Guanidinated casein inhibited the solubilized BBM binding to alpha-casein more strongly than casein (casein sodium and alpha-casein) inhibited. Trypsinization of solubilized BBM abolished its binding activity to alpha-casein. These results indicate that some membrane protein, but not aminopeptidases, contained in BBM interacts with dietary proteins, and that guanidinated casein has a higher affinity for BBM than intact casein. These binding intensities for proteins were closely correlated to physiological responsiveness, and are possibly involved in a sensory system for dietary protein in the intestine. PMID- 11440111 TI - Relationship between the enzymatic browning and phenylalanine ammonia-lyase activity of cut lettuce, and the prevention of browning by inhibitors of polyphenol biosynthesis. AB - Cut lettuce stored at 4 degrees C gradually turned brown on the cut section after several days of storage. Three factors for enzymatic browning, the polyphenol content, polyphenol oxidase activity, and phenylalanine ammonia-lyase (PAL) activity, were examined during the cold storage of cut lettuce. A relationship between the browning and PAL activity was apparent. We tried to prevent this browning by using the two enzyme inhibitors, 2-aminoindane-2-phosphonic acid (AIP), an inhibitor of the phenylpropanoid pathway, and glyphosate, an inhibitor of the shikimate pathway. AIP and glyphosate significantly inhibited the browning of cut lettuce. The polyphenol content and PAL activity were both reduced by the treatment with AIP. These results show that regulating the biosynthesis of polyphenols is essential to prevent the browning of cut lettuce. PMID- 11440112 TI - Concentration of docosahexaenoic acid from fish oils using Geotrichum sp. FO347 2. AB - Geotrichum sp. FO347-2 could use refined sardine oil as a sole carbon source. Dry cell mass reached a maximum of 0.788 g per g of the oil added for 72 h. Total weight of the cellular lipids was largest around 24 h, when the contents of triglyceride and free fatty acid were 63.6 and 22.2%, respectively. Docosahexaenoic acid (DHA) was incorporated and concentrated in the cellular lipids, and the content reached 25.9% for 24 h, adding sardine oil containing 12.3% DHA. DHA and eicosapentaenoic acid were also concentrated in residual lipids outside the cells. Using tuna head oil containing 26.8% DHA, FO347-2 was compared with Candida guilliermondii FO726A with respect to DHA incorporation. FO347-2 and FO726A accumulated similar amounts of DHA, i.e. 53 and 55 mg, respectively, in 1 g of freeze-dried cells after 24-h cultivation at 30 degrees C. The recovery rates of DHA from the tuna oil for FO347-2 and FO726A were 19.4 and 19.7%, respectively. PMID- 11440113 TI - Production of beta-thujaplicin in Cupressus lusitanica suspension cultures fed with organic acids and monoterpenes. AB - Effects of some organic acids and monoterpenes on production of beta-thujaplicin were studied in Cupressus lusitanica suspension cultures. The fungal elicitor induced biosynthesis of beta-thujaplicin was promoted by the feedings of malate, pyruvate, fumarate, succinate, and acetate. These results suggest some relationships between acetate/pyruvate metabolism and beta-thujaplicin biosynthesis, or between tricarboxylic acid cycle and beta-thujaplicin biosynthesis. Feedings of C. lusitanica suspension cultures with some monoterpenes inhibited elicitor-triggered beta-thujaplicin biosynthesis, but 2 carene and terpinyl acetate feedings significantly improved the beta-thujaplicin production of C. lusitanica suspension cultures. These results indicate a possible involvement of terpinyl acetate and 2-carene in beta-thujaplicin biosynthesis, as well as potential uses of these monoterpenes in large-scale beta thujaplicin production. PMID- 11440114 TI - Rapid degradation of the silkworm diapause hormone by trypsin and its suppression by VAP-map, a synthetic analog of the cuticular peptide of silkworm, Bm ACP-6.7 (VAP-peptide). AB - Very fast tryptic degradation of the silkworm diapause hormone was found and the degradation pathway was analyzed by moderating the reaction conditions. It proceeded via cleavage at Arg23 and finally at Arg15 of DH. As the C-terminal structure of DH was essential for exhibiting bioactivity, the first cleavage caused rapid inactivation of the hormone. This tryptic digestion was strongly suppressed by adding VAP-map, a synthetic analog of the cuticular peptide of silkmoths, Bm ACP-6.7 (VAP-peptide), which is a natural synergist of DH. VAP-map suppressed the enzymic reaction by interacting with the substrate, but not with the enzyme. PMID- 11440115 TI - Isolation and characterization of the novel lipophilic protein, Pb CP-12.7, from the shell of the pink shrimp, Pandalus borealis. AB - Past research on diapause-inducing substances of the silkworm has isolated an extremely lipophilic peptide and demonstrated its unique characteristics. In the present work, similar lipophilic proteins were searched for in the shell of the shrimp, Pandalus borealis, and one novel protein, Pb CP-12.7, was isolated. Its structure comprising 126 amino acids was revealed by a combination of a sequence analysis and the enzymic fragmentation technique. Pb CP-12.7 is unique in that it was insoluble in neutral-slightly basic water, but highly soluble in some organic solvents. It contained an abundance of hydrophobic amino acids and repeating sequences. In addition, it was adsorbed to chitin, a major component of the shell of the shrimp. PMID- 11440116 TI - Behavior of pesticides (1): efficiency evaluation of pesticides. AB - The correlation equation for the concentration-mortality curve of a specific pesticide and a control pesticide was used to adapt the effectiveness data from residual activity tests to the corresponding chemical application concentration, to calculate the constant with the decreasing speed equation, and to calculate the application dose necessary to maintain an equal residual effect period by the same equation. The calculated value is the amount of pesticide needed to perform with the same efficiency as the control pesticide (effectiveness and period of effectiveness), which is vitally important to effectively select an appropriate pesticide. PMID- 11440117 TI - Molecular cloning of acid-stable glucose isomerase gene from Streptomyces olivaceoviridis E-86 by a simple two-step PCR method, and its expression in Escherichia coli. AB - Glucose isomerase (GI) from Streptomyces olivaceoviridis E-86 is a unique enzyme, very acid-stable with a large potential for corn sweetener industries. The gene encoding this unique enzyme was cloned by a simple two-step PCR method, and expressed in Escherichia coli. A single open reading frame consisting of 1164 base pairs (70.7 mol % of G + C content) that encoded a polypeptide composed of 388 amino acid residues (Mr 42,993) was found. The E. coli transformant carrying the gene overproduced the recombinant GI (rGI) and the enzyme was successfully expressed as a tetramer under the transcriptional control of the tac-promoter. The purified recombinant enzyme was indistinguishable from that of the authentic enzyme e.g. molecular weight, immunological properties, N-terminal amino acid sequences, subunit structures, and temperature and pH profiles. The relationships between structure and properties of the enzymes are also discussed. PMID- 11440118 TI - Suppression of collagen-induced arthritis in DBA/1J mice by preimmunization with house dust mite extract. AB - Collagen-induced arthritis (CIA) can be induced in DBA/1J mice by immunization with bovine type II collagen (bCII) and is a model of some types of human autoimmune rheumatoid arthritis. In this study we examined whether preimmunization of the mice with various antigens could inhibit the development of CIA. Preimmunization of the mice with an extract of the house dust mite Dermatophagoides farinae (mite antigen), chicken ovalbumin, or keyhole limpet hemocyanin strongly inhibited CIA development, but hen egg lysozyme, beta lactoglobulin from bovine milk or myelin basic protein from guinea pig brain did not substantially affect CIA development. Splenic T cells and serum antibodies specific for mite antigen did not cross-react with bCII. Preimmunization of the mice with mite antigen did not affect the IFN-gamma and proliferative response of splenic T cells to bCII, nor serum antibody responses. The most inhibitory constituent had a molecular weight between 1,000 and 10,000. PMID- 11440119 TI - Reduction by phytate-reduced soybean beta-conglycinin of plasma triglyceride level of young and adult rats. AB - This study examined the effects of soybean beta-conglycinin, from which phytate was mostly removed, on the plasma lipids in young and adult rats. Male Wistar young (6 week-old) and adult (21 week-old) rats were fed high cholesterol diets containing 20% casein, soy protein isolate (SPI), or soybean beta-conglycinin for 10 days. In young rats, although the food intake of the beta-conglycinin group was higher than those of the casein and SPI groups, the weight gain was significantly lower than those of the other groups. However, in adult rats, the weight gain was not different among the groups. In young and adult rats, relative liver weights of SPI and beta-conglycinin groups were significantly lower than that of the casein group, and the degree of the reduction was more marked in the beta-conglycinin group than in the SPI group. In young rats, the plasma triglyceride level was significantly lower in the SPI and beta-conglycinin groups than that in the casein group. In addition, the plasma triglyceride level of the beta-conglycinin group was significantly lower than that of the SPI group. Plasma total cholesterol levels of the SPI and beta-conglycinin groups were significantly lower than that of the casein group. However, there was little difference in the lowering effect between SPI and beta-conglycinin. These results indicate that soybean beta-conglycinin may have lowering functions not only on plasma total cholesterol level, but also on plasma triglyceride level. PMID- 11440120 TI - Labeling patterns of chloroplastidic isoprenoids in cultured cells of liverwort Ptychanthus striatus. AB - Incorporation studies administering 2H- and 13C-labeled mevalonate (MVA) and 13C labeled glucose to suspension cultured cells of the liverwort, Ptychanthus striatus, were carried out in order to examine the biosynthesis of the phytyl side-chain of chlorophyll a. Administration of 13C- and 2H-labeled MVA provided evidence for the involvement of the MVA pathway in the phytyl side-chain biosynthesis and preferential labeling of the farnesyl diphosphate (FPP)-derived portion. An alternate labeling pattern in the phytyl side-chain was observed which was slightly different to the non-equivalent labeling in other liverworts, such as Heteroscyphus planus and Lophocolea heterophylla and in the hornwort, Anthoceros punctatus. The labeling pattern observed after the administration of 13C-labeled glucose revealed the simultaneous involvement of the non-MVA pathway in the phytol biosynthesis of P. striatus cells. PMID- 11440121 TI - Production of humanized antibody against human high-affinity IgE receptor in a serum-free culture of CHO cells, and purification of the Fab fragments. AB - We describe the preparation of Fab fragments of a humanized anti-human high affinity IgE receptor (Fc epsilonRIalpha) antibody potentially useful for treatment of IgE-mediated allergic diseases. IgE-binding capacities of sixteen combinations of light and heavy chains of four recombinant anti-Fc epsilonRIalpha antibodies, chimeric CRA2, humanized CRA2, chimeric CRA4, and humanized CRA4, were compared. A combination in which both chains were of humanized CRA2 had the highest activity. Stable transfectant clones of four kinds of host cells expressing recombinant antibodies were established. CHO-K1 cells were the most productive. Serum-free media suitable for culture of the stable CHO-transfectant clones were screened. The concentration of the humanized CRA2, which the most productive clone secreted into the chosen serum-free medium, was approximately 100 microg/ml. A procedure for the purification of the antibody, papain digestion, and purification of Fab fragments was established. The highly purified humanized Fab fragments are suitable for use to examine their in vivo activity and immunogenicity in primates. PMID- 11440122 TI - Digested bacterial cell powder (DBCP) as a source of reduced-form folates for pigs: use of a trimethoprim-resistant strain and the bioavailability of folates in DBCP. AB - The production of digested bacterial cell powder (DBCP) as a source of reduced form folates for pigs was studied. Trimethoprim-resistant mutants of Brevibacterium lactofermentum ATCC 13869 accumulated a significantly higher amount of the reduced form of folate in the cells than the wild-type strain. DBCPs were prepared from the resistant mutant strain and the wild-type strain. The utilization of the reduced-form of folate in DBCP was evaluated by measuring the plasma folate level after orally administering DBCP to Gottingen minipigs. The folates in both DBCPs proved to have equally high bioavailability in the pigs. PMID- 11440123 TI - Purification and characterization of a cold-adapted isocitrate lyase and a malate synthase from Colwellia maris, a psychrophilic bacterium. AB - Isocitrate lyase (ICL) and malate synthase (MS) of a psychrophilic marine bacterium, Colwellia maris, were purified to electrophoretically homogeneous state. The molecular mass of the ICL was found to be 240 kDa, composed of four identical subunits of 64.7 kDa. MS was a dimeric enzyme composed of 76.3 kDa subunits. N-Terminal amino acid sequences of the ICL and MS were analyzed. Purified ICL had its maximum activity at 20 degrees C and was rapidly inactivated at the temperatures above 30 degrees C, but the optimum temperature for the activity of MS was 45 degrees C. NaCl was found to protect ICL from heat inactivation above 30 degrees C, but the salt did not stabilize MS. Effects of temperatures on the kinetic parameters of both the enzymes were examined. The Km for the substrate (isocitrate) of ICL was decreased with decreasing temperature. On the other hand, the Km for the substrate (glyoxylate) of MS was increased with decreasing temperature. The calculated value of free energy of activation of ICL was on the same level as that of MS. PMID- 11440124 TI - New biological function of bovine alpha-lactalbumin: protective effect against ethanol- and stress-induced gastric mucosal injury in rats. AB - Although several studies have shown that milk protein components have a wide range of biological activities, the potential role of these proteins in the gastrointestinal mucosal defense system is less well elucidated. In this study, we investigated the effect of the major proteins in cow's milk on gastric mucosal injury by using two acute ulcer models in Wistar rats. Gastric mucosal injury was induced by either intragastric 60% ethanol-HCl or water-immersion restraint stress (23 degrees C, 7 h). Each test milk protein was orally administered 30 min before the induction of gastric injury. Among the major milk proteins, alpha lactalbumin (alpha-LA) is demonstrated to have a marked protective effect against ethanol-induced gastric injury, with the same potency as that of the typical antiulcer agent, Selbex. Whey protein isolate (WPI), which contained 25% alpha LA, also protected against gastric injury, while casein showed no effect. Comparative studies on the protective effect of the four major components of WPI, beta-lactoglobulin, alpha-LA, bovine serum albumin and gamma-globulins (immunoglobulins), on the basis of their contents in WPI revealed that alpha-LA was responsible for the protective effect of WPI, being about 4-fold more effective than WPI itself. Alpha-LA showed dose-dependent protection against gastric injury induced by stress as well as ethanol. Pretreatment with indomethacin (10 mg/kg body weight, s.c.), which is a potent inhibitor of endogenous prostaglandin synthesis, resulted in a significant reduction in the protective effect of alpha-LA. These results indicate that alpha-LA has marked antiulcer activity as an active component of cow's milk protein, and suggest that alpha-LA intake may serve to protect against gastric mucosal injury, in part through endogenous prostaglandin synthesis. PMID- 11440125 TI - Adenine deaminase activity of the yicP gene product of Escherichia coli. AB - During previous work on deriving inosine-producing mutants of Escherichia coli, we observed that an excess of adenine added to the culture medium was quickly converted to hypoxanthine. This phenomenon was still apparent after disruption of the known adenosine deaminase gene (add) on the E. coli chromosome, suggesting that, like Bacillus subtilis, E. coli has an adenine deaminase. As the yicP gene of E. coli shares about 35% identity with the B. subtilis adenine deaminase gene (ade), we cloned yicP from the E. coli genome and developed a strain that overexpressed its product. The enzyme was purified from a cell extract of E. coli harboring a plasmid containing the cloned yicP gene, and had significant adenine deaminase [EC 3.5.4.2] activity. It was deduced to be a homodimer, each subunit having a molecular mass of 60 kDa. The enzyme required manganese ions as a cofactor, and adenine was its only substrate. Its optimum pH was 6.5-7.0 and its optimum temperature was 60 degrees C. The apparent Km for adenine was 0.8 mM. PMID- 11440126 TI - Revised sequence and expression of cyclin B cDNA from the starfish Asterina pectinifera. AB - Cyclin B cDNA was cloned from the ovary of the starfish Asterina pectinifera and analyzed by RT-PCR and 3'- and 5'-RACE techniques. The cDNA consists of a 0.13-kb upstream untranslated region, a 1.22-kb coding region, and a 0.86-kb downstream untranslated region. The open reading frame encoded a polypeptide of 404 amino acid residues with a calculated molecular weight of 45,692. All the characteristic sequences, such as destruction and cyclin boxes, cyclin B motif, and cytoplasmic retention and nuclear export signals, were found in the newly cloned cyclin B cDNA. The deduced amino acid sequence of the cyclin B cDNA was highly homologous in the middle and carboxy terminal regions to that from mature eggs of the same organism, but quite different in the amino terminal region. Evidence was obtained which suggested that this cyclin B is expressed in immature and maturing oocytes and is the same as that cloned from mature eggs. PMID- 11440127 TI - Fused heterocyclic antioxidants: antioxidative activities of hydrocoumarins in a homogeneous solution. AB - We compared the antioxidative activities of seven hydrocoumarins with those of alpha-tocopherol for the oxidation of tetralin and linoleic acid in a homogeneous solution. Hydrocoumarins exhibited a higher induction period than that of alpha Toc in both systems. However, the rate of oxygen absorption during the induction period for alpha-Toc was slower than that of the hydrocoumarins in both systems. In addition, 6,7-dihydroxy-4,4-dimethylhydrocoumarin showed less cytotoxicity toward human fibroblasts than did 2,6-di-t-butyl-4-methylphenol. PMID- 11440128 TI - Stereoselective synthesis of the neolignan, (+ )-dehydrodiconiferyl alcohol. AB - A stereo controlled synthesis of the biologically active neolignan, (+) dehydrodiconiferyl alcohol (1) was achieved. This synthetic method was also efficient for preparing its enantiomer and other derivatives with biological activity. PMID- 11440129 TI - Major isoforms of starch branching enzymes in premature seeds of kidney bean (Phaseolus vulgaris L.). AB - Developing seeds of the kidney bean (Phaseolus vulgaris L.) contain several isoforms of starch branching enzymes. Two of them, KBE1 and KBE2, which are the major forms in the premature seeds, were purified as a single band of protein on SDS-PAGE and native PAGE by chromatographies on DEAE-Sepharose, Bio-Gel P-200, and amylose-binding Sepharose 6B. The enzymes had similar pH optimum (7.0), pH stability (7.0-9.5), temperature optimum (25-30 degrees C), and temperature stability (up to 40 degrees C). Additionally, both were inhibited by various divalent metal ions and activated by citrate. Finally, though their N-terminal amino acid sequences were identical, their molecular masses and affinities for amylose differed; 80 kDa and 1.27 mM for KBE1 and 77 kDa and 0.74 mM for KBE2. PMID- 11440130 TI - Mutation analysis of the feedback inhibition site of aspartokinase III of Escherichia coli K-12 and its use in L-threonine production. AB - Aspartokinase III (AKIII), one of three isozymes of Escherichia coli K-12, is inhibited allosterically by L-lysine. This enzyme is encoded by the lysC gene and has 449 amino acid residues. We analyzed the feedback inhibition site of AKIII by generating various lysC mutants in a plasmid vector. These mutants conferred resistance to L-lysine and/or an L-lysine analogue on their host. The inhibitory effects of L-lysine on and heat tolerance of 14 mutant enzymes were examined and DNA sequencing showed that the types of mutants were 12. Two hot spots, amino acid residue positions 318-325 and 345-352, were detected in the C-terminal region of AKIII and these enzyme regions may be important in L-lysine-mediated feedback inhibition of AKIII. Feedback resistant lysC relieved on L-threonine hyper-producing strain, B-3996, from reduced L-threonine productivity by addition of L-lysine, and furthermore increased L-threonine productivity even when no addition of L-lysine. It suggested that the bottleneck of L-threonine production of B-3996 was AK and feedback resistant lysC was effective because of the strict inhibition by cytoplasmic L-lysine. PMID- 11440131 TI - Arginine as an exacerbating factor for glomerulonephritis in rats fed a methionine-threonine-supplemented low casein diet. AB - We previously demonstrated that a methionine-threonine-supplemented low (8.5%) casein diet (8.5CMT) reduced symptoms such as proteinuria in nephritic rats without severe protein malnutrition. In this study, we examined whether or not L arginine supplementation to 8.5CMT would exacerbate proteinuria and other symptoms in nephritic rats. Male Wistar rats with glomerulonephritis induced by a single intravenous injection of nephrotoxic serum were fed either a 20% casein diet (control), 8.5% casein diet, 8.5CMT, or L-arginine-supplemented 8.5CMT (8.5CMTA) for 16 days. The 8.5CMTA, as compared with the 8.5CMT, aggravated proteinuria and glomerulonephritis. Administration of L-N(G)-nitroarginine methyl ester, an inhibitor of nitric oxide synthase, to 8.5CMTA-fed nephritic rats by drinking water for 14 days canceled the adverse effect of L-arginine on proteinuria and histopathological damage in glomeruli. These results suggest that the supplementation of L-arginine makes exacerbation via nitric oxide production in glomerulonephritis. PMID- 11440132 TI - Purification and characterization of ginsenoside Rb1-metabolizing beta glucosidase from Fusobacterium K-60, a human intestinal anaerobic bacterium. AB - Fusobacterium K-60, a ginsenoside Rb1-metabolizing bacterium, was isolated from human intestinal feces. From this Fusodobacterium K-60, a ginsenoside Rb1 metabolizing enzyme, beta-glucosidase, has been purified. The enzyme was purified to apparent homogeneity by a combination of butyl-Toyopearl, hydroxyapatite ultragel, Q-Sepharose, and Sephacryl S-300 HR column chromatographies with a final specific activity of 1.52 micromol/min/mg. It had optimal activity at pH 7.0 and 40 degrees C. The molecular mass of this purified enzyme was 320 kDa, with 4 identical subunits (80 kDa). The purified enzyme activity was inhibited by Ba++, Fe++, and some agents that modify cysteine residues. This enzyme strongly hydrolyzed sophorose, followed by p-nitrophenyl beta-D-glucopyranoside, esculin, and ginsenoside Rb1. However, this enzyme did not change 20-O-beta-D glucopyranosyl-20(S)-protopanaxadiol (IH-901) to 20(S)-protopanaxadiol, while it weakly changed ginsenoside Rb1 to IH-901. These findings suggest that the Fusobacterial beta-glucosidase is a novel enzyme transforming ginsenoside Rb1. PMID- 11440133 TI - Apoptosis of antigen-specific T cells induced by oral administration of antigen: comparison of intestinal and non-intestinal immune organs. AB - Oral administration of a protein without adjuvant brings about oral tolerance (systemic hyporesponsiveness) to that protein by mechanisms such as antigen induced apoptosis. We monitored the number and apoptosis induction of CD4+ T cells in antigen-specific T cell receptor transgenic mice fed the antigen ovalbumin to identify where events leading to oral tolerance occurred. The antigen was distributed throughout the body, causing apoptosis and a decrease in cell number of CD4+ T cells in most of the lymphoid system: the spleen, peripheral lymph nodes, and the thymus which was not previously reported to be affected. Although apoptosis was induced in the Peyer's patches, the cell number did not change. Unexpectedly, T cells in the mesenteric lymph nodes did not undergo apoptosis; instead, they were more numerous as compared to that in the case of control animals not administered the antigen. The results suggested that the orally administered antigen activated the intestinal immune system, while it induced immune tolerance in other sites. PMID- 11440134 TI - Cloning and characterization of the cpyA gene encoding intracellular carboxypeptidase from Aspergillus nidulans. AB - Carboxypeptidase Y (CPY) has been used as a maker enzyme for investigations on intracellular transport of vacuolar proteins and on vacuolar biogenesis in Saccharomyces cerevisiae. We describe the cloning and characterization of the CPY homologue encoding gene (cpyA) from the filamentous fungus Aspergillus nidulans. The cpyA gene has one intron and encodes a protein with 552 amino acids containing a putative signal sequence and pro-sequence. The predicted CpyA protein is highly similar in sequence with carboxypeptidases from several yeast species and contains a catalytic triad (Asp-His-Ser) like that of serine carboxypeptidase. The cpyA disruptant cells showed reduced levels of intracellular carboxypeptidase. These results suggest that the cpyA gene encodes a vacuolar carboxypeptidase in A. nidulans. PMID- 11440135 TI - A fibronectin-binding protein from rice bran with cell adhesion activity for animal tumor cells. AB - A rice bran 57-kDa protein was isolated by affinity chromatography with fibronectin immobilized on agarose. This fibronectin-binding protein designated as RB-57 had an amino-terminal amino acid sequence identical with that of a putative mature form of rice hydroxyproline-rich glycoprotein. A distinct feature of the amino acid composition of RB-57 was the high contents of hydroxyproline and proline representing about 45% of the total amino acids. The sugar analysis indicated that arabinose represented 46.8% of the total carbohydrates. RB-57 showed cell adhesion activity for murine Lewis lung carcinoma cells. The result suggests that RB-57 may play a role in plant cell adhesion, although cell adhesion-promoting activity for plant cells remains to be tested. PMID- 11440136 TI - Phosphopeptides derived from hen egg yolk phosvitin: effect of molecular size on the calcium-binding properties. AB - Two different phosphopeptide (PPP) fragments derived from partially dephosphorylated hen egg yolk phosvitin were prepared by tryptic digestion, and their Ca2+ binding property compared with that of commercial casein phosphopeptides (CPP). The smaller fragment of less than 1 kDa and O-phospho-1 serine did not bind Ca2+ to any significant extend, while PPP of 1-3 kDa showed a higher ability than CPP to render soluble calcium. The results show that not only the phosphoserine residues are critical for Ca2+ binding, but also the molecular size of the phosphopeptides. PMID- 11440137 TI - Isolation of poly(3-hydroxybutyrate) (PHB)-degrading microorganisms and characterization of PHB-depolymerase from Arthrobacter sp. strain W6. AB - Microbial degraders of poly(3-hydroxybutyrate) (PHB) were isolated from soil. Arthrobacter sp. strain W6 used not only PHB as a carbon source, but also PHAs such as poly(3-hydroxybutyrate-co-[5%]3-hydroxyvalerate), poly(3-hydroxybutyrate co-[14%]3-hydroxyvalerate), and poly(3-hydroxybutyrate-co-[22%]3 hydroxyvalerate). PHB-depolymerase was purified to homogeneity from the culture broth of Arthrobacter sp. strain W6 by a procedure involving DEAE- and butyl Toyopearl column chromatographies. The Mr of the enzyme was estimated to be about 47,000 by SDS-polyacrylamide gel electrophoresis. The enzyme was most active at pH 8.5 and 50 degrees C, and was inhibited by phenylmethylsulfonyl fluoride, Hg2+, Ag+, and Pb2+. PMID- 11440138 TI - Aurantiamide acetate, a selective cathepsin inhibitor, produced by Aspergillus penicilloides. AB - Aurantiamide acetate was isolated from the fermentation broth of Aspergillus penicilloides for the first time. Aurantiamide acetate inhibited cysteine proteinases, in particular, cathepsin L (3.4.22.15) and B (3.4.22.1) with IC50 of 12 microM and 49 microM, respectively. In the adjuvant-arthritic rat model, subcutaneously administered 10 mg/kg body weight of this compound suppressed hind paw swelling. PMID- 11440139 TI - Lupeol esters from the twig bark of Japanese pear (Pyrus serotina Rehd.) cv. Shinko. AB - Five new lupeol esters, lup-20(29)-ene-3beta-yl eicosanoate, docosanoate, tetracosanoate, hexacosanoate and octacosanoate, were isolated as a mixture from the twig bark of Japanese pear (Pyrus serotina Rehd.) cv. Shinko, together with lupeol and epifriedelinol. Their structures were determined by spectral analyses including 2D-NMR experiments. PMID- 11440140 TI - Introduction of a low molecular weight agonist peptide for complement C3a receptor into soybean proglycinin A1aB1b subunit by site-directed mutagenesis. AB - LPLPR, a complement C3a agonist peptide, with hypocholesterolemic activity was introduced into the homologous site of soybean proglycinin A1aB1b subunit by site directed mutagenesis. This modified proglycinin was expressed in E. coli and recovered from the insoluble fraction. LPLPR was released by the action of chymotrypsin and trypsin as expected. Furthermore, two peptides (RPSYLPLPR and PSYLPLPR) with extended sequence at the amino-terminus of LPLPR were obtained. Their ileum-contracting activity was 9 to approximately 13 times stronger than that of LPLPR. The overall yields of purified LPLPR, RPSYLPLPR and PSYLPLPR were 25%, 12%, and 0.7% respectively. PMID- 11440141 TI - A novel C1-using denitrifier alcaligenes sp. STC1 and its genes for copper containing nitrite reductase and azurin. AB - A novel denitrifier Alcaligenes sp. STC1 was identified. The strain efficiently denitrifies under an atmosphere of 10% oxygen (O2) where Paracoccus denitrificans, one of the most studied aerobic denitrifiers, had less denitrifying activity, indicating that the strain has an O2-torelant denitrifying system. It denitrified by using C1-carbon sources such as formate and methanol as well as glucose, glycerol, and succinate. The genes for the copper-containing nitrite reductase and azurin of this C1-using denitrifier were cloned. Their predicted products of them were similar to those of their counterparts and the maximum similarities were 90% and 92%, respectively. PMID- 11440142 TI - Glycitein effect on suppressing the proliferation and stimulating the differentiation of osteoblastic MC3T3-E1 cells. AB - Glycitein, as one of the three major isoflavones in soybeans, directly but significantly (about 5%) suppressed the proliferation of MC3T3-E1 and stimulated bone-related protein (alkaline phosphatase (ALP) and osteocalcin (OC)) expression. These results indicate that glycitein suppresses the proliferation of osteoblasts and promotes differentiation from its progenitor. PMID- 11440143 TI - Estimation of the number of polyhydroxyalkanoate (PHA)-degraders in soil and isolation of degraders based on the method of most probable number (MPN) using PHA-film. AB - A new method to estimate the number of polyhydroxyalkanoates (PHA)-degraders in soil and to isolate degraders, called the film-MPN method, is proposed. The incubation time was measured by the first order reaction (FOR) model. This method was used to estimate numbers of poly(3-hydroxybutyrate-co-3 hydroxyvalerate)[P(3HB-co-3HV)]- and poly(3-hydroxyvalerate-co-4 hydroxybutyrate)[P(3HB-co-4HB)]-degraders in garden soil (4.30 x 10(5) and 2.15 x 10(5) aerobic degraders per gram of dry soil, respectively). The number of P(3HB co-3HV)-degraders in paddy field soil was 5.06 x 10(5) aerobic degraders per gram dry soil. Also, several P(3HB-co-3HV)-degraders were isolated directly from positive-growth tubes of high dilution. PMID- 11440144 TI - Flagellin as a biomarker for Bacillus subtilis strains; application to the DB9011 strain and the study of interspecific diversity in amino-acid sequences. AB - Polymerase chain reaction (PCR) for the flagellin central domain coding region (FCD-PCR) was applied to the detection and discrimination of Bacillus subtilis DB9011, a strain with useful functions in agriculture. Cross-reactions were observed in 4 B. subtilis strains with similar flagellin genes (hag). Alignment of partial amino-acid sequences of flagellin and the results of PCR for the 16S/23S rRNA spacer in 11 B. subtilis strains suggested the presence of a group including strains with antifungal activity (DB9011 and others). PMID- 11440145 TI - Enzymatic synthesis of two ginsenoside Re-beta-xylosides. AB - Two new beta-xylosyl derivatives of ginsenoside Re, 20(S)-protopanaxatriol 6-O alpha-L-rhamnopyranosyl-(1 --> 2)-[beta-D-xylopyranosyl-(1 --> 4)]-beta-D glucopyranosyl-20-O-beta-D-glucopyranoside and 20(S)-protopanaxatriol 6-O-alpha-L rhamnopyranosyl-(1 --> 2)-[beta-D-xylopyranosyl-(1 --> 6)]-beta-D-glucopyranosyl 20-O-beta-D-glucopyranoside, were respectively synthesized from p-nitrophenyl beta-D-xylopyranoside and phenyl beta-D-xylopyranoside as donors and ginsenoside Re as the acceptor in 25% acetone and acetonitrile by a cellulase preparation from Trichoderma viride and a beta-galactosidase preparation from Aspergillus oryzae. The latter enzyme preparation also catalyzed the hydrolysis of ginsenoside Re to the minor saponin, ginsenoside Rg2. PMID- 11440146 TI - Role of lipophilicity and hydrogen peroxide formation in the cytotoxicity of flavonols. AB - We compared the lipophilicity and toxicity of the four flavonols, galangin, kaempferol, quercetin and myricetin, which respectively have no, one, two and three hydroxyl groups on the B-ring. The lipophilicity was in the order of myricetin < quercetin < kaempferol < galangin. The cytotoxicity determined by a colony-formation assay with Chinese hamster lung fibroblast V79 cells was in the order of quercetin < kaempferol < galangin < myricetin. Apart from myricetin, the order of lipophilicity was the same as that of cytotoxicity, implying that the cytotoxicity was attributable to the lipophilicity. The cytotoxicity of myricetin was attributable to the hydrogen peroxide formed by autoxidation. PMID- 11440147 TI - gsk disruption leads to guanosine accumulation in Escherichia coli. AB - We tried some improvement of inosine production using an inosine-producing mutant of Escherichia coli which is deficient in purF (phosphoribosylpyrophosphate (PRPP) amidotransferase gene), purA (succinyl-adenosine 5'-monophosphate (AMP) synthetase gene), deoD (purine nucleoside phosphorylase gene), purR (purine repressor gene) and add (adenosine deaminase gene), and harboring the desensitized PRPP amidotransferase gene as a plasmid. The guaB (inosine 5' monophosphate (IMP) dehydrogenase gene) disruption brought about a slightly positive effect on the inosine productivity. Alternatively, the gsk (guanosine inosine kinase gene) disruption caused a considerable amount of guanosine accumulation together with a slight increase in the inosine productivity. The further addition of guaC (guanosine 5'-monophosphate (GMP) reductase gene) disruption did not lead to an increased guanosine accumulation, but brought about the decrease of inosine accumulation. PMID- 11440148 TI - Visualization of biallelic expression of the imprinted SNRPN gene induced by inhibitors of DNA methylation and histone deacetylation. AB - We have adapted a fluorescence in situ hybridization (FISH) method to detect nascent RNA molecules of the imprinted SNRPN gene at the initial transcription sites in nuclei of human HL60 and WI38 cells. Simultaneous detection of RNA and DNA of SNRPN by FISH using the cosmid probe confirmed its monoallelic expression in these cell lines. Treatment of both cells by inhibitors of DNA methylation and histone deacetylation resulted in time-dependent increase of the cell population with the biallelic expression of SNRPN. PMID- 11440149 TI - Efficient synthesis of O-linked glycopeptide by a transglycosylation using endo alpha-N-acetylgalactosaminidase from Streptomyces sp. AB - Gal beta-(1-->3)-GalNAc-linked hexapeptide was synthesized by a transglycosylation using Gal beta-(1-->3)-GalNAc beta-pNP as a donor and a serine containing hexapeptide as an acceptor using endo GalNAc-ase from Streptomyces sp.. The Gal beta-(1-->3)-GalNAc residue was transferred to the hydroxyl group of the serine residue of the peptide. The total yield of the glycopeptide via this process was better than that of the chemoenzymatic method. This process was confirmed to be a versatile method for the synthesis of O-linked glycopeptides. PMID- 11440150 TI - Total synthesis of (+)-albicanol and (+)-albicanyl acetate. AB - The drimane sesquiterpenes, (+)-albicanol (2) and (+)-albicanyl acetate (3), were synthesized from an optically active bicyclic diol [(+)-1] that had been obtained via the recently developed optical resolution of a general synthetic intermediate for drimane sesquiterpenes. The crucial step in the previous syntheses was markedly improved by the modified Wittig methylenation of a silyloxy ketone (7). The high overall yield (77% in 4 or 5 steps from (+)-1) by this total synthesis makes it possible to synthesize the other biologically active drimane sesquiterpenes. PMID- 11440151 TI - Preparation of extended metaphase chromosomes from human cultured cells using a topoisomerase II inhibitor, ICRF-193. AB - Effects of ICRF-193, a topoisomerase II inhibitor, on metaphase chromosome preparations were examined. A short-time exposure of this drug to human HL60 cells in a suspension culture before harvest resulted in obtaining more extended metaphase chromosomes. The length of chromosome 6 identified by fluorescence in situ hybridization was twice as long with this drug treatment. Together with effectiveness for adherent HepG2 cells, these results suggest that treatments with ICRF-193 provide a simple and reliable method for extended metaphase chromosome preparations from cultured cells. PMID- 11440152 TI - Pentalenolactone I and hygromycin A, immunosuppressants produced by Streptomyces filipinensis and Streptomyces hygroscopicus. AB - Immunosuppressants isolated from Streptomyces filipinensis and S. hygroscopicus were identified with pentalenolactone I and hygromycin A, respectively. The compounds as well as cyclosporin A showed immunosuppressant activity in the mixed lymphocyte reaction, and pentalenolactone I and cyclosporin A suppressed IL-2 production, however, hygromycin A did not. Hygromycin A may have immunosuppressant activity by a different mechanism from pentalenolactone I, cyclosporin A and tacrolimus. PMID- 11440153 TI - Identification of N-terminal autodigestion target site in subtilisin ALP I. AB - Autodigestion of subtilisin ALP I (ALP I), secreted from the alkalophilic Bacillus sp. NKS-21 and its predicted amino acid sequence having about 60% identity with other alkaline subtilisins, was examined under alkaline conditions. At the alkaline pH of 12, ALP I was rapidly degraded, and almost no breakdown products were detectable. However, by incubating ALP I at 5 degrees C for an extended time, a couple of specific peptides (26.7 kDa and 25.6 kDa) were accumulated. Each of them was purified and amino acid sequences of these fragments were found. Both peptides appeared to start at Gly-19 of the mature sequence of ALP I. PMID- 11440154 TI - Microbial resolution of 2-hydroxy-3-nitropropionic acid for synthesis of optically active isoserine. AB - The biocatalytic stereoselective hydrolysis of 2-hydroxy-3-nitropropionic acid esters was studied. Forty enzymes and three hundred microorganism strains were examined for their ability to hydrolyze ethyl 2-hydroxy-3-nitropropionic acid. Nocardia globerula IFO13150 gave n-butyl (R)-2-hydroxy-3-nitropropionate with a 92% enantiomeric excess (ee) and the corresponding carboxylic acid with a 92%ee, which was easily converted to (S)-isoserine, a useful beta-amino acid. PMID- 11440155 TI - Features of distamycin preferential binding sites on natural DNA predicted using differential scanning calorimetry. AB - The interaction of distamycin with ColE1 DNA was examined by using differential scanning calorimetry (DSC) taking the helix-coil transition theory of DNA into consideration. Our results here strongly indicate that the affinity of distamycin to DNA, at a low distamycin concentration, depends highly on the DNA sequence, and preferential binding occurs to the sites of four to six successive A-T pairs having two or more successive G-C pairs on both their ends. PMID- 11440156 TI - Evidence for the presence of DNA-binding proteins involved in regulation of the gene expression of indole-3-pyruvic acid decarboxylase, a key enzyme in indole-3 acetic acid biosynthesis in Azospirillum lipoferum FS. AB - We isolated the ipdc gene coding for indole-3-pyruvic acid decarboxylase (IPDC), a key enzyme in the indole-3-pyruvic acid pathway for indole-3-acetic acid biosynthesis, in the plant growth-promoting rhizobacterium Azospirillum lipoferum FS. Gel mobility-shift assay showed the presence of two DNA-binding proteins that might be involved in regulation of the ipdc gene expression. PMID- 11440157 TI - Expression of PR-5d and ERF genes in cultured tobacco cells and their NaCl stress response. AB - The expression of pathogenesis-related protein-5d (PR-5d) and ethylene-responsive transcriptional factors (ERF1-4) for basic PR-proteins was measured in cultured tobacco cells. The PR-5d transcript markedly increased with culture, but only constant levels of transcripts of ERF1 (trans-activator) and ERF3 (repressor) were detected in cells. These different responses of PR-5d and ERF genes were also observed under NaCl stress; PR-5d transcript decreased and ERF levels were constant under NaCl stress. The effect of post-transcriptional regulation of ERF activity, especially ERF3, on the expression of basic PR genes including PR-5d is discussed. PMID- 11440158 TI - Structural studies of cysteine proteases and their inhibitors. AB - Cysteine proteases (CPs) are responsible for many biochemical processes occurring in living organisms and they have been implicated in the development and progression of several diseases that involve abnormal protein turnover. The activity of CPs is regulated among others by their specific inhibitors: cystatins. The main aim of this review is to discuss the structure-activity relationships of cysteine proteases and cystatins, as well as of some synthetic inhibitors of cysteine proteases structurally based on the binding fragments of cystatins. PMID- 11440159 TI - Structure and dynamics of a DNA duplex containing single alpha-anomeric deoxyadenosine residue. AB - Structure and dynamics of an undecamer DNA duplex containing a single alpha anomeric deoxyadenosine residue placed in opposition to a thymidine unit have been studied using simulation of molecular dynamics in aqueous solution. Despite several noticeable deviations from the B-DNA duplex structure caused by the anomerisation, such as: West type puckering of the alpha-anomeric sugar, disrupted base stacking pattern and unstable duplex bending, the formation of a non-classical alpha-dA-T pair was observed. A novel way of visual presentation of trajectory data allowing high throughput screening of the conformational parameters is presented. PMID- 11440160 TI - Cation binding properties of calretinin, an EF-hand calcium-binding protein. AB - Calretinin (CR) is a neuronal EF-hand protein previously characterized as a calcium (micromolar affinity) binding protein. CR-containing neurons are spared in some neurodegenerative diseases, although it is as yet unconfirmed how CR plays an active role in this protection. Higher levels of some metal cations (e.g. copper and zinc) are associated with these diseases. At the same time, metals such as terbium (NMR and fluorescence) cadmium (NMR) and manganese (EPR) serve as useful calcium analogues in the study of EF-hand proteins. We survey the binding of the above-mentioned metal cations that might affect the structure and function of CR. Competitive 45Ca2+-overlay, competitive terbium fluorescence and intrinsic tryptophan fluorescence are used to detect the binding of metal cations to CR. Terbium and copper (half-maximal effect of 15 microM) bind to CR. Terbium has a similar or greater affinity for the calcium-binding sites of CR than calcium. Copper quenches the fluorescence of terbium-bound CR, and CR tryptophan residues and competes weakly for 45Ca2+-binding sites. Cadmium, magnesium, manganese and zinc bind less strongly (half-maximal effects above 0.1 mM). Therefore, only terbium appears to be a suitable analytical calcium analogue in further studies of CR. The principal conclusion of this work is that copper, in addition to calcium, might be a factor in the function of CR and a link between CR and neurodegenerative diseases. PMID- 11440161 TI - Immunosuppressory activity of the cyclodimeric peptide with RGD-sequences. AB - Our previous studies showed that the nonapeptide fragment of HLA-DQ of the sequence H-Thr-Pro-Gln-Arg-Gly-Asp-Val-Tyr-Thr-OH, located in the beta164-172 loop, strongly suppresses the humoral and cellular immune responses, while its shorter analogs, H-Arg-Gly-Asp-Val-OH, H-Arg-Gly-Asp-Val-Tyr-OH and H-Gln-Arg-Gly Asp-Val-Tyr-OH show only a weak stimulatory activity in respect to the humoral immunological response. These fragments contain the Arg-Gly-Asp (RGD) sequence, known for its importance for cellular association phenomena. Based on the crystal structure of HLA-DR1, we also designed and synthesized a cyclic analog H-Cys-Arg Gly-Asp-Val-Tyr-Cys-OH with restricted conformation, which strongly suppresses the immune response and selectively inhibits the alphavbeta3 integrin, suggesting that the mechanism of the immunosuppressory action of the peptide is associated with inhibition of the integrin. In this paper we present the design and synthesis of the cyclodimeric peptide, Arg-Gly-Asp-Arg-Gly-Asp, which is also known as a selective alphavbeta3 inhibitor. The synthesized peptide strongly suppresses both the humoral and cellular immune response. The results support our hypothesis that the immunomodulatory activity of HLA-DQ fragments may be connected with their interactions with some particular integrins on the cell surface. PMID- 11440162 TI - Molecular docking-based test for affinities of two ligands toward vasopressin and oxytocin receptors. AB - Molecular docking simulations are now fast developing area of research. In this work we describe an effective procedure of preparation of the receptor-ligand complexes. The amino-acid residues involved in ligand binding were identified and described. PMID- 11440163 TI - Intracellular potassium and chloride channels: an update. AB - Channels selective for potassium or chloride ions are present in all intracellular membranes such as mitochondrial membranes, sarcoplasmic/endoplasmic reticulum, nuclear membrane and chromaffin granule membranes. They probably play an important role in events such as acidification of intracellular compartments and regulation of organelle volume. Additionally, intracellular ion channels are targets for pharmacologically active compounds, e.g. mitochondrial potassium channels interact with potassium channel openers such as diazoxide. This review describes current observations concerning the properties and functional roles of intracellular potassium and chloride channels. PMID- 11440164 TI - Uncoupling proteins in mitochondria of plants and some microorganisms. AB - Uncoupling proteins, members of the mitochondrial carrier family, are present in mitochondrial inner membrane and mediate free fatty acid-activated, purine nucleotide-inhibited H+ re-uptake. Since 1995, it has been shown that the uncoupling protein is present in many higher plants and some microorganisms like non-photosynthetic amoeboid protozoon, Acanthamoeba castellanii and non fermentative yeast Candida parapsilosis. In mitochondria of these organisms, uncoupling protein activity is revealed not only by stimulation of state 4 respiration by free fatty acids accompanied by decrease in membrane potential (these effects being partially released by ATP and GTP) but mainly by lowering ADP/O ratio during state 3 respiration. Plant and microorganism uncoupling proteins are able to divert very efficiently energy from oxidative phosphorylation, competing for deltamicroH+ with ATP synthase. Functional connection and physiological role of uncoupling protein and alternative oxidase, two main energy-dissipating systems in plant-type mitochondria, are discussed. PMID- 11440165 TI - Mitochondrial uncoupling does not influence the stability of the intracellular signal activating plasma membrane calcium channels. AB - The effects of various concentrations of thapsigargin, a specific inhibitor of Ca2+-ATPase in the endoplasmic reticulum (ER) membrane, on calcium homeostasis in lymphoidal T cells (Jurkat) were investigated. Preincubation of these cells suspended in nominally calcium-free medium with 0.1 microM thapsigargin resulted in a complete release of Ca2+ from intracellular calcium stores. When the medium was supplemented with 3 mM CaCl2 the cells maintained constantly elevated level of cytosolic Ca2+. However, thapsigargin applied at lower concentration produced only a partial depletion of the stores. For example, in the cells pretreated with 1 nM thapsigargin and suspended in calcium-free medium approximately 75% of the calcium content was released from the intracellular stores. The addition of 3 mM CaCl2 to such cell suspension led to a transient increase in cytosolic calcium concentration, followed by a return to a lower steady-state. This phenomenon, related to the refilling of the ER by Ca2+, allowed to estimate the half-time for the process of cell recovery after activation of store-operated calcium channels. By this approach we have found that carbonyl cyanide m-chlorophenylhydrazone, which has been documented to inhibit calcium entry into Jurkat cells, does not influence the stability of the intracellular signal involved in the activation of store-operated calcium channels. PMID- 11440166 TI - Translocation of polysialic acid across model membranes: kinetic analysis and dynamic studies. AB - Transmembrane translocation of polyion homopolymers takes place in the case of polyanionic polysialic acid (polySia), polyanionic polynucleotides and polycationic polypeptides. The purpose of this work was to determine the role of membrane electrical parameters on the kinetics of polyion translocation, the influence of polysialic acid on ion adsorption on positively charged membrane surface and the dynamics of the phospholipid hydrocarbon chains and choline group by using 1H-NMR. The analysis of polyion translocation was performed by using the electrical equivalent circuit of the membrane for the initial membrane potential equal to zero. The changes in polysialic acid flux was up to 75% after 1 ms in comparison with the zero-time flux. Both a decrease of membrane conductance and an increase of polyion chain length resulted in the diminution of this effect. An increase of praseodymium ions adsorption to positively charged liposomes and an increase of the rate of segmental movement of the -CH2 and -CH3 groups, and the choline headgrup of lipid molecules, was observed in the presence of polySia. The results show that the direction of the vectorial polyion translocation depends both on the membrane electrical properties and the degree of polymerization of the polymer, and that polysialic acid can modulate the degree of ion adsorption and the dynamics of membrane lipids. PMID- 11440167 TI - Signal transduction in confluent C3H 10T1/2 cells. The role of focal adhesion kinase. AB - The activities of extracellular signal-regulated kinases (ERK1/ERK2) is required for proliferation of several types of cells. The performed analysis showed stimulation of ERK's by fetal calf serum (FCS) or fibronectin in the C3H 10T1/2 cell cultures at logarithmic phase of growth. The ERKs activity was not stimulated in confluent cells. This could not be accounted for a partial down regulation of ERK since its level was stable in both types of cells regardless of their density and kind of stimulation. Searching for ERK up-stream elements we studied the integrin receptor gene transcript by RT-PCR and focal adhesion kinase (FAK) by Western blotting and phosphorylation assays. It was found that FCS and fibronectin stimulated phosphorylating activity of FAK in the cells at the logarithmic phase of growth, but were inefficient in the confluent cells. RT-PCR showed the presence of alpha5 and beta1 integrin transcripts, and p125FAK was at the same level regardless of the type of stimulation. These data indicate that the ability of FAK to be activated plays an important role in ERK regulation and, in consequence in proliferation and growth inhibition during confluence. PMID- 11440168 TI - The flavonoids, quercetin and isorhamnetin 3-O-acylglucosides diminish neutrophil oxidative metabolism and lipid peroxidation. AB - Two natural flavonoids, quercetin and isorhamnetin 3-O-acylglucosides, were examined for their inhibitory influence on the in vitro production and release of reactive oxygen species in polymorphonuclear neutrophils (PMNs). The generation of superoxide radical, hydrogen peroxide and hypochlorous acid were measured by, respectively, cytochrome c reduction, dichlorofluorescin oxidation and taurine chlorination. Membrane lipid oxidation was studied by the thiobarbituric acid method in mouse spleen microsomes. The addition of flavonoids at the concentration range 1-100 microM inhibited PMNs oxidative metabolism and lipid peroxidation in a dose-dependent manner. The results suggest that these flavonoids suppress the oxidative burst of PMNs and protect membranes against lipid peroxidation. PMID- 11440169 TI - Structure and functions of 5S rRNA. AB - The ribosome is a macromolecular assembly that is responsible for protein biosynthesis in all organisms. It is composed of two-subunit, ribonucleoprotein particles that translate the genetic material into an encoded polypeptides. The small subunit is the site of codon-anticodon interaction between the messenger RNA (mRNA) and transfer RNA (tRNA) substrates, and the large subunit catalyses peptide bond formation. The peptidyltransferase activity is fulfilled by 23S rRNA, which means that ribosome is a ribozyme. 5S rRNA is a conserved component of the large ribosomal subunit that is thought to enhance protein synthesis by stabilizing ribosome structure. This paper shortly summarises new results obtained on the structure and function of 5S rRNA. PMID- 11440170 TI - Dendrimers: properties and applications. AB - Dendrimers are a new class of polymeric materials. They are highly branched, monodisperse macromolecules. The structure of these materials has a great impact on their physical and chemical properties. As a result of their unique behaviour dendrimers are suitable for a wide range of biomedical and industrial applications. The paper gives a concise review of dendrimers' physico-chemical properties and their possible use in various areas of research, technology and treatment. PMID- 11440171 TI - On the relationship between fractal geometry of space and time in which a system of interacting cells exists and dynamics of gene expression. AB - We report that both space and time, in which a system of interacting cells exists, possess fractal structure. Each single cell of the system can restore the hierarchical organization and dynamic features of the entire tumor. There is a relationship between dynamics of gene expression and connectivity (i.e., interconnectedness which denotes the existence of complex, dynamic relationships in a population of cells leading to the emergence of global features in the system that would never appear in a single cell existing out of the system). Fractal structure emerges owing to non-bijectivity of dynamic cellular network of genes and their regulatory elements. It disappears during tumor progression. This latter state is characterized by damped dynamics of gene expression, loss of connectivity, loss of collectivity (i.e., capability of the interconnected cells to interact in a common mode), and metastatic phenotype. Fractal structure of both space and time is necessary for a cellular system to self-organize. Our findings indicate that results of molecular studies on gene expression should be interpreted in terms of space-time geometry of the cellular system. In particular, the dynamics of gene expression in cancer cells existing in a malignant tumor is not identical with the dynamics of gene expression in the same cells cultured in the monolayer system. PMID- 11440172 TI - Application of genetic semihomology algorithm to theoretical studies on various protein families. AB - Several protein families of different nature were studied for genetic relationship, correct alignment at non-homologous fragments, optimal sequence consensus construction, and confirmation of their actual relevance. A comparison of the genetic semihomology approach with statistical approaches indicates a high accuracy and cognition significance of the former. This is particularly pronounced in the study of related proteins that show a low degree of homology. The sequence multiple alignments were verified and corrected with respect to the questionable, non-homologous fragments. The verified alignments were the basis for consensus sequence formation. The frequency of six-codon amino acids occurrence versus position variability was studied and their possible role in amino acid mutational exchange at variable positions is discussed. PMID- 11440173 TI - A novel isoform of human lymphoid enhancer-binding factor-1 (LEF-1) gene transcript encodes a protein devoid of HMG domain and nuclear localization signal. AB - Lymphoid enhancer-binding factor-1 (LEF-1), a member of the high mobility group (HMG) family of proteins, regulates expression of T-cell receptor-alpha gene and is one of the key regulatory molecules in the epithelial-mesenchymal interactions during embryonic development. Among others, LEF-1 regulates expression of cytokeratin genes involved in formation of hair follicles and the gene encoding the cell-adhesion molecule E-cadherin. Transcription factor LEF-1, which acts as a dimer, binds beta-catenin and is involved in signal transduction by the wnt pathway. We have cloned and sequenced a novel isoform of human LEF-1 gene transcript. This isoform encodes a truncated protein devoid of HMG domain and nuclear localization signal but retaining beta-catenin binding domain. This isoform might either act in a dominant-negative manner by interfering with native LEF-1, or might bind beta-catenin in the cytosol, which would result in attenuation of the signals transmitted by the LEF-beta-catenin pathway. PMID- 11440174 TI - Expression of bovine leukemia virus protein p24 in Escherichia coli and its use in the immunoblotting assay. AB - The gag gene encoded protein, p24 of bovine leukemia virus (BLV), was cloned and expressed as thioredoxin-6xHis-p24 protein in Escherichia coli. The bacterial cells carrying plasmid pT7THis-p24 expressed the protein of 38 kDa that was detected by immunoblotting analysis using anti-p24 monoclonal antibodies and sera from BLV infected cattle and sheep. The purified p24 fusion protein was shown to be sensitive and specific for detection of BLV antibodies in the infected cattle. PMID- 11440175 TI - Protective action of vitamin C against DNA damage induced by selenium-cisplatin conjugate. AB - Genotoxicity of anticancer drugs is of a special interest due to the risk of inducing secondary malignancies. Vitamin C (ascorbic acid) is a recognized antioxidant and, since human diet can be easily supplemented with vitamin C, it seems reasonable to check whether it can protect against DNA-damaging effects of antitumor drugs. In the present work the ability of vitamin C to modulate cytotoxic and genotoxic effects of a cisplatin analog, conjugate (NH3)2Pt(SeO3), in terms of cell viability, DNA damage and repair in human lymphocytes was examined using the trypan blue exclusion test and the alkaline comet assay, respectively. The conjugate evoked a concentration-dependent decrease in the cell viability, reaching nearly 50% at 250 microM. (NH3)2Pt(SeO3) at 1, 10 and 30 microM caused DNA strand breaks, measured as the increase in the comet tail moment of the lymphocytes. The treated cells were able to recover within a 30-min incubation in a drug-free medium at 37 degrees C. Vitamin C at 10 and 50 microM diminished the extent of DNA damage evoked by (NH3)2Pt(SeO3) but had no effect on the kinetics of DNA repair. The vitamin did not directly inactivate the conjugate. Lymphocytes treated with endonuclease III, which recognises oxidised pyrimidines, displayed a greater tail moment than those untreated with the enzyme, suggesting that the damages induced by the drug have, at least in part, an oxidative origin. Vitamin C can be considered a potential protective agent against side effects of antitumor drugs, but further research with both normal and cancer cells are needed to clarify this point. PMID- 11440176 TI - Therapeutical effect of modified adamantane copolymer compounds: study of molecular mechanisms. AB - Copolymers of N-polyvinylpyrrolidone-acrylic acid (AB-1) and adamantane derivatives are known to possess marked antiviral activity in in vitro and in ovo models. Among the constructed preparations of AB-1 modified by adamantane derivatives some, especially AB-4 (modified by deitiforin), were found to show more extended antiviral activity and to inhibit markedly virus reproduction in susceptible permissive cell cultures and chicken embryos. In AB-4 treated cells and allantoic sacs, virus titers (influenza virus, herpes virus, and HIV) and virus antigen concentration were decreased. On the other hand, herpes virus specific thymidine kinase and of DNA-polymerases isolated from Escherichia coli, Plectonema boryanum, and herpes virus type 1 infected murine brain tissue retained their activity after incubation with AB-4 or AB-2. The compounds investigated, in view of their effect on virus reproduction, are thought to be prospective as antiviral agents. PMID- 11440177 TI - Modulation of human deoxycytidine kinase activity as a response to cellular stress induced by NaF. AB - Deoxycytidine kinase (dCK) is one of the key enzymes of deoxynucleoside salvage supplying resting lymphocytes with DNA precursors for synthesis and repair. The level of dCK activity is especially important in chemotherapy with the use of deoxynucleoside analogues like arabinosyl cytosine (Citarabid, ara-C), or 2 chloro-deoxyadenosine (Cladribine, CdA). Previous results showed that Cladribine treatment of human lymphocytes increased several fold the activity of dCK without increasing the amount of dCK protein itself (Sasvari-Szekely, et al., 1998, Biochem. Pharmacol. 56, 1175), and a possible post-translational modification was suggested. This theory was further investigated using NaF as an inhibitor of protein phosphatases. It was shown that NaF treatment of cells elevated dCK activity while inhibiting DNA synthesis. The possible mechanism of dCK activation/inactivation induced by exposure of cell cultures to different agents is discussed. PMID- 11440178 TI - The interactions of phthalocyanines with stimulated and resting human peripheral blood mononuclear cells. AB - The interactions of two metal-free phthalocyanines [(H2Pc) and Solar Pc (with four peripherical groups: SO2N(CH2CH2OH)2)] and of one metal substituted dye (CoPc) with resting and stimulated human peripheral blood mononuclear cells (PBMC) were compared. The absorption, fluorescence, photoacoustic and EPR spectra of both resting cells and cells stimulated by phytohaemagglutinin, incubated in dimethyl sulfoxide (DMSO) with very low or 95% water content and with or without dye addition, were measured. The fate of the light absorbed by the samples was investigated. It is known that singlet oxygen production is crucial for photodynamic action of dyes. Thermal deactivation and luminescence emission compete with this process, so investigation of these alternative paths of sensitizer deactivation provides information about photodynamic action. The incorporation of the investigated dyes into cells and the perturbation of the cell structure caused by the dyes, the incubation solvent and the activator were investigated by comparing the spectral properties of PBMC before and after stimulation and incubation. Incubation of the cells for 1 h in a solution of Solar Pc in 99.5% aqueous DMSO, resulted in an efficient dye incorporation which was highly selective. Solar Pc being introduced much more efficiently into stimulated cells than into resting cells. PMID- 11440179 TI - Chlorination of N-acetyltyrosine with HOCl, chloramines, and myeloperoxidase hydrogen peroxide-chloride system. AB - N-acetyl-L-tyrosine (N-acTyr), with the alpha amine residue blocked by acetylation, can mimic the reactivity of exposed tyrosyl residues incorporated into polypeptides. In this study chlorination of N-acTyr residue at positions 3 and 5 in reactions with NaOCl, chloramines and the myeloperoxidase (MPO)-H2O2-Cl- chlorinating system were invesigated. The reaction of N-acTyr with HOCl/OCl- depends on the reactant concentration ratio employed. At the OCl-/N-acTyr (molar) ratio 1:4 and pH 5.0 the chlorination reaction yield is about 96% and 3 chlorotyrosine is the predominant reaction product. At the OCl-/N-acTyr molar ratio 1:1.1 both 3-chlorotyrosine and 3,5-dichlorotyrosine are formed. The yield of tyrosine chlorination depends also on pH, amounting to 100% at pH 5.5, 91% at pH 4.5 and 66% at pH 3.0. Replacing HOCl/OCl- by leucine/chloramine or alanine/chloramine in the reaction system, at pH 4.5 and 7.4, produces trace amount of 3-chlorotyrosine with the reaction yield of about 2% only. Employing the MPO-H2O2-Cl- chlorinating system at pH 5.4, production of a small amount of N acTyr 3-chloroderivative was observed, but the reaction yield was low due to the rapid inactivation of MPO in the reaction system. The study results indicate that direct chlorination of tyrosyl residues which are not incorporated into the polypeptide structure occurs with excess HOCl/OCl- in acidic media. Due to the inability of the myeloperoxidase-H2O2-Cl- system to produce high enough HOCl concentrations, the MPO-mediated tyrosyl residue chlorination is not effective. Semistable amino-acid chloramines also appeared not effective as chlorine donors in direct tyrosyl chlorination. PMID- 11440180 TI - Tritolylporphyrin dimer as a new potent hydrophobic sensitizer for photodynamic therapy of melanoma. AB - We report the synthesis, photochemical and photophysical properties and preliminary studies on biological effect of a new tritolylporphyrin dimer (T-D). Absorption and emission properties of T-D suggest its possible use in photodynamic therapy. T-D is capable of singlet oxygen production with 0.8 quantum yield. It also has a high photostability. The photodynamic properties of the dimer were examined following the growth of SKMEL 188 (human melanoma) cells irradiated with red light (cut off < 630 nm). The surviving fraction of the cells decreased about 3-fold (vs. non-irradiated cells) for an 81 J/cm dose. Our results suggest that tritolylporphyrine dimer T-D may be an interesting hydrophobic sensitizer for photodynamic therapy. PMID- 11440181 TI - Antioxidant status in erythrocytes of cystic fibrosis children. AB - Activities of superoxide dismutase, catalase and glutathione peroxidase in erythrocytes of cystic fibrosis children were studied in order to estimate the severity of their deficiency. Our results point to increased susceptibility of erythrocytes of cystic fibrosis subjects to oxidative injury and indicate that the antioxidant status of patients should be carefully monitored. PMID- 11440182 TI - Theoretical models of catalytic domains of protein phosphatases 1 and 2A with Zn2+ and Mn2+ metal dications and putative bioligands in their catalytic centers. AB - The oligomeric metalloenzymes protein phosphatases dephosphorylate OH groups of Ser/Thr or Tyr residues of proteins whose actions depend on the phosphorus signal. The catalytic units of Ser/Thr protein phosphatases 1, 2A and 2B (PP1c, PP2Ac and PP2Bc, respectively), which exhibit about 45% sequence similarity, have their active centers practically identical. This feature strongly suggests that the unknown structure of PP2Ac could be successfully homology-modeled from the known structures of PP1c and/or PP2Bc. Initially, a theoretical model of PP1c was built, including a phosphate and a metal dication in its catalytic site. The latter was modeled, together with a structural hydroxyl anion, as a triangular pseudo-molecule (Zno or Mno), composed of two metal cations (double Zn2+ or Mn2+, respectively) and the OH- group. To the free PP1c two inhibitor sequences R29RRRPpTPAMLFR40 of DARPP-32 and R30RRRPpTPATLVLT42 of Inhibitor-1, and two putative substrate sequences LRRApSVA and QRRQRKpRRTI were subsequently docked. In the next step, a free PP2Ac model was built via homology re-modeling of the PP1c template and the same four sequences were docked to it. Thus, together, 20 starting model complexes were built, allowing for combination of the Zno and Mno pseudo-molecules, free enzymes and the peptide ligands docked in the catalytic sites of PP1c and PP2Ac. All models were subsequently subjected to 250-300 ps molecular dynamics using the AMBER 5.0 program. The equilibrated trajectories of the final 50 ps were taken for further analyses. The theoretical models of PP1c complexes, irrespective of the dication type, exhibited increased mobilities in the following residue ranges: 195-200, 273-278, 287-209 for the inhibitor sequences and 21-25, 194-200, 222-227, 261, 299-302 for the substrate sequences. Paradoxically, the analogous PP2Ac models appeared much more stable in similar simulations, since only their "prosegment" residues 6-10 and 14-18 exhibited an increased mobility in the inhibitor complexes while no areas of increased mobility were found in the substrate complexes. Another general observation was that the complexes with Mn dications were more stable than those with Zn dications for both PP1c and PP2Ac units. PMID- 11440183 TI - Novel approach to computer modeling of seven-helical transmembrane proteins: current progress in the test case of bacteriorhodopsin. AB - G-protein coupled receptors (GPCRs) are thought to be proteins with 7-membered transmembrane helical bundles (7TM proteins). Recently, the X-ray structures have been solved for two such proteins, namely for bacteriorhodopsin (BR) and rhodopsin (Rh), the latter being a GPCR. Despite similarities, the structures are different enough to suggest that 3D models for different GPCRs cannot be obtained directly employing 3D structures of BR or Rh as a unique template. The approach to computer modeling of 7TM proteins developed in this work was capable of reproducing the experimental X-ray structure of BR with great accuracy. A combination of helical packing and low-energy conformers for loops most close to the X-ray structure possesses the r.m.s.d. value of 3.13 A. Such a level of accuracy for the 3D-structure prediction for a 216-residue protein has not been achieved, so far, by any available ab initio procedure of protein folding. The approach may produce also other energetically consistent combinations of helical bundles and loop conformers, creating a variety of possible templates for 3D structures of 7TM proteins, including GPCRs. These templates may provide experimentalists with various plausible options for 3D structure of a given GPCR; in our view, only experiments will determine the final choice of the most reasonable 3D template. PMID- 11440184 TI - The effect of the Glu342Lys mutation in alpha1-antitrypsin on its structure, studied by molecular modelling methods. AB - The structure of native alpha1-antitrypsin, the most abundant protease inhibitor in human plasma, is characterised primarily by a reactive loop containing the centre of proteinase inhibition, and a beta-sheet composed of five strands. Mobility of the reactive loop is confined as a result of electrostatic interactions between side chains of Glu342 and Lys290, both located at the junction of the reactive loop and the beta structure. The most common mutation in the protein, resulting in its inactivation, is Glu342-->Lys, named the Z mutation. The main goal of this work was to investigate the influence of the Z mutation on the structure of alpha1-antitrypsin. Commonly used molecular modelling methods have been applied in a comparative study of two protein models: the wild type and the Z mutant. The results indicate that the Z mutation introduces local instabilities in the region of the reactive loop. Moreover, even parts of the protein located far apart from the mutation region are affected. The Z mutation causes a relative change in the total energy of about 3%. Relatively small root mean square differences between the optimised structures of the wild type and the Z mutant, together with detailed analysis of 'conformational searching' process, lead to the hypothesis that the Z mutation principally induces a change in the dynamics of alpha1-antitrypsin. PMID- 11440185 TI - Monte carlo simulations of protein-like heteropolymers. AB - Properties of a simple model of polypeptide chains were studied by the means of the Monte Carlo method. The chains were built on the (310) hybrid lattice. The residues interacted with long-range potential. There were two kinds of residues: hydrophobic and hydrophilic forming a typical helical pattern -HHPPHPP-. Short range potential was used to prefer helical conformations of the chain. It was found that at low temperatures the model chain formes dense and partially ordered structures (non-unique). The presence of the local potential led to an increase of helicity. The effect of the interplay between the two potentials was studied. After the collapse of the chain further annealing caused rearrangement of helical structures. Dynamic properties of the chain at low temperature depended strongly on the local chain ordering. PMID- 11440186 TI - Molecular modelling study of the role of cholesterol in the stimulation of the oxytocin receptor. AB - Cholesterol, an integral component of membranes in Eucaryota, is a modifier of membrane properties. In vivo studies have demonstrated that cholesterol can also modulate activities of some G protein-coupled receptors (GPCRs), which are integral membrane proteins. This can result either from an effect of cholesterol on the membrane fluidity or from specific interactions of the membrane cholesterol with the receptor, as recently demonstrated for the cholecystokinin type beta (CCKRbeta) or the oxytocin receptor (OTR). Using molecular modelling, we studied conformational preferences of cholesterol and several of its analogues. Subsequently, we simulated the distributions of their preferred conformations around the surface of OTR, CCKRbeta and a chimeric oxytocin/cholecystokinin receptor. Consequently, we suggest residues on the surface of OTR which are potentially significant in the OTR/cholesterol interaction. PMID- 11440187 TI - Fluorescence decay time distribution analysis of cyclic enkephalin analogues; influence of solvent and Leu configuration in position 5 on conformation. AB - Lifetime distribution analysis were performed to study the influence of Leu configuration in position 5 on changes of the peptide chain of cyclic analogues of enkephalins containing a fluorescence donor and acceptor in different solvents. The configuration change of Leu5 in all the analogues of enkephalins studied which contain donor-acceptor pairs has no apparent influence on Trp lifetime distributions. In contrast, there is a significant solvent effect on the shape of lifetime distribution. PMID- 11440188 TI - Immunosuppressive effects of aflatoxin B1 in Indian major carp (Labeo rohita). AB - Aflatoxin B1 (AFB1) is a potent immunomodulator in endotherms. The experiment was carried out to study the immunosuppressive nature of AFB1 in one ectothermic species of Indian major carp. Graded levels (0, 1.25, 5.00 mg/kg of body weight) of purified AFB1 were intraperitoneally (i.p.) injected into rohu (Labeo rohita) fingerlings weighing 30-50 g, and the fish were observed for a period of 90 days. At the end of the trial, blood samples were collected from the control group as well as the AFB1 injected fish and were screened for nitroblue tetrazolium (NBT) assay, serum total protein, albumin, globulin, albumin-globulin ratio (A:G), serum bactericidal activity and bacterial agglutination titre against Edwardsiella tarda. The aflatoxin-treated fish revealed a reduction of total protein, globulin levels, bacterial agglutination titre, NBT and serum bactericidal activities, as well as an enhanced A:G ratio without change in albumin concentration, irrespective of dose levels of toxin treatment, when compared to the control group. Thus, AFB1 proved to be immunosuppressive in rohu even at the lowest dose (1.25 mg/kg body weight) of toxin treatment. This could be of economic significance in intensive culture systems of rohu. PMID- 11440189 TI - Differential effects of heparin on NO and tumor necrosis factor-alpha production in bovine blood mononuclear cells stimulated with Salmonella typhimurium lipopolisaccharide. AB - We investigated the influence of heparin, one of the extracellular matrix (ECM) components, on nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) production by bovine peripheral blood mononuclear cells (PBMC) and monocytes left to adhere for 2 (freshly adherent monocytes) and 48 h (resting monocytes), activated with Salmonella typhimurium lipopolysaccharide (LPS). After 24-h stimulation with LPS, heparin (100 microg/ml) increased (by about 40%) NO production by peripheral blood mononuclear cells and by freshly adherent monocytes. However, it did not change NO synthesis by the resting monocytes. Unlike its influence on NO level, heparin diminished TNF-alpha production by PBMC and monocytes stimulated with LPS. Microscopical examination of PBMC stained with biotin-labeled heparin, showed that both lymphocytes and monocytes were able to bind this glycosaminoglycan. We suggest that heparin, as a component of ECM, modulates the early response of monocytes to exogenous stimuli. PMID- 11440190 TI - Chronic Fatigue Syndrome (CFS) in 15 dogs and cats with specific biochemical and microbiological anomalies. AB - A great deal of controversy and speculation surrounds the etiology of Chronic Fatigue Syndrome (CFS) in human patients and the existence of a similar illness in animals. To evaluate the association with a presumptive staphylococcal infection and bacteremia, seven dogs and eight cats diagnosed with CFS (two meeting the CDC working case definition) were submitted to rapid blood cultures and fresh blood smears investigations. Nine out of 15 blood cultures proved Staph positive and four isolates were specified as S. xilosus (3) and S. intermedius (1). The presence of micrococci-like organisms in the blood was of common observation among these subjects, in association with fatigue/pain-related symptoms and biochemical abnormalities suggestive of a myopathy. Following treatment with a low dosage arsenical drug (thiacetarsamide sodium, Caparsolate, i.v., 0.1 ml/kg/day) all patients experienced complete remission. Micrococci disappeared from the blood at post-treatment controls made 10-30 days later. The outcomes were compared with those of five healthy controls and five 'sick with other illness' patients showing significant difference. PMID- 11440191 TI - Nitric oxide production by macrophages of dogs vaccinated with killed Leishmania infantum promastigotes. AB - Human visceral leishmaniosis is endemic in Southern Italy, where the dog is the main reservoir of viscerotropic strains of Leishmania infantum. The release of nitric oxide (NO) by interferon (IFN)-gamma-activated macrophages is an important leishmanicidal mechanism in several animal species. In this work NO production, phagocytosis and killing capacity of monocyte-derived dog macrophages were evaluated in vitro before and after administration of a vaccine composed of killed Leishmania infantum promastigotes. Moreover, IFN-gamma content was measured in concanavalin A-activated dog peripheral blood mononuclear cell (PBMC) supernatants employed for macrophage stimulation. Phagocytosis, killing capacity and NO production by canine macrophages increased significantly 1 month after vaccine administration, and the increase also persisted 5 months later. In addition, the amount of IFN-gamma in PBMC supernatants was significantly higher after vaccination. Overall, our results suggest the usefulness of evaluating the in vivo protective role of this promastigote preparation in dogs. PMID- 11440192 TI - Analysis of IgG response to experimental infection with RH Toxoplasma gondii in goats. AB - The IgG response of goats experimentally infected with RH Toxoplasma gondii has been analysed using an indirect ELISA and Western-blot analysis. Specific IgG antibodies were first detected at 14 days post-inoculation (p.i.), reaching a peak by day 35 p.i. and showing slight fluctuations until the end of the experiment (91 p.i.). Specific IgG showed a reactivity over a whole range of peptides (125-24 kDa approximately), but the highest reactivity was observed against a group of antigens with a molecular weight between 34 and 28 kDa, in particular against a 30 kDa fraction which is considered to represent the major surface protein of T. gondii named p30 or SAG-1. PMID- 11440193 TI - Intracellular calcium changes in neuronal cells induced by Alzheimer's beta amyloid protein are blocked by estradiol and cholesterol. AB - 1. The elevation of intracellular Ca2+ levels ([Ca2+]i) in immortalized hypothalamic neurons (GT1-7 cells) after exposure to Alzheimer's beta-amyloid protein (AbetaP[25-35]) was investigated using a multisite fluorometry system. 2. The marked rise in [Ca2+]i appeared after exposure to 5-20-microM AbetaP[25-35]. Analysis of the spatiotemporal patterns of [Ca2+]i changes revealed that the magnitude and the latency of the response to AbetaP in each cell were highly heterogeneous. 3. The preadministration of 17beta-estradiol, 17alpha-estradiol, phloretin and cholesterol, which influence the properties of membranes, such as membrane fluidity or membrane potential, significantly decreased the rise in [Ca2+]i. 4. These findings support the idea that disruption of calcium homeostasis by AbetaP channels may be the molecular basis of the neurotoxicity of AbetaP and of the pathogenesis of Alzheimer's disease. It is also suggested that membrane properties may play key roles in the expression of neurotoxicity. PMID- 11440194 TI - Coupling of nerve growth factor to its receptor: inhibition by anti-GM3 ganglioside antibody. AB - 1. Normal differentiation of PC 12 cells and dorsal root ganglionic neurons in culture need nerve growth factor (NGF) for their neurite outgrowth. 2. An antibody against GM3 ganglioside was found to inhibit the nerve growth factor mediated neurite formation of both the cells in vitro significantly. 3. Further analysis revealed that the binding of 125I-NGF to live PC 12 cells could be markedly inhibited by anti-GM3 antibody in a dose dependent manner. 4. Scatchard analysis revealed that in the presence of anti-GM3 antibody only some low affinity binding sites were available for NGF-high affinity binding sites were totally blocked. 5. These results further strengthen the hypothesis that anti-GM3 antibody affects neuronal cell growth by interfering with the coupling of growth factors to their cell surface receptors. PMID- 11440195 TI - Diabetes increases the expression of hypothalamic neuropeptides in a spontaneous model of type I diabetes, the nonobese diabetic (NOD) mouse. AB - 1. Synthesis of oxytocin (OT) and arginine-vasopressin (AVP) is increased in induced models of Type I diabetes, such as the streptozotocin model. However, these parameters have not yet been evaluated in spontaneous models, such as the nonobese diabetic mouse (NOD). Therefore, we studied in the magnocellular cells of the paraventricular nucleus (PVN) of nondiabetic and diabetic 16-week-old female NOD mice and control C57B1/6 mice, the immunocytochemistry of OT and AVP peptides and their mRNA expression, using nonisotopic in situ hybridization (ISH). 2. In nondiabetic and diabetic NOD female mice, the number of OT- and AVP immunoreactive cells were similar to those of the controls, whereas immunoreaction intensity was significantly higher for both peptides in diabetic NOD as compared with nondiabetic NOD and control C57B1/6 mice. 3. ISH analysis showed that the number of OT mRNA-containing cells was in the same range in the three groups, whereas higher number of AVP mRNA expressing cells was found in diabetic NOD mice. However, the intensity of hybridization signal was also higher for both OT and AVP mRNA in the diabetic group as compared with nondiabetic NOD and control mice. 4. Blood chemistry demonstrated that haematrocrit, total plasma proteins, urea, sodium, and potassium were within normal limits in diabetic mice. Thus, NOD mice were neither hypernatremic nor dehydrated. 5. We suggest that upregulation of OT and AVP reflects a high-stress condition in the NOD mice. Diabetes may affect neuropeptide-producing cells of the PVN, with the increased AVP and OT playing a deleterious role on the outcome of the disease. PMID- 11440196 TI - Subcellular localization of fragile X mental retardation protein with the I304N mutation in the RNA-binding domain in cultured hippocampal neurons. AB - 1. Fragile X syndrome, the most common form of inherited mental retardation, is caused by the lack or dysfunction of fragile X mental retardation protein (FMRP). The 1304N mutation in the RNA-binding domain of FMRP results in an exceptionally severe form of mental retardation. 2. We have investigated the subcellular localization of FMRP and its 1304N-mutated form in cultured hippocampal neurons and PC12 cells, using immunofluorescence microscopy. In PC12 cells, FMRP was predominantly localized to the cytoplasm and also to the processes after differentiation by NGF. 3. In cultured hippocampal neurons, granular labeling was detected along the neuronal processes. 4. Double-labeling with synaptophysin antibody revealed FMRP at synaptic sites in neurons. 5. The 1304N mutation did not appear to affect the transport of FMRP to dendrites or its localization at synaptic sites. Thus, FMRP is a synaptic protein and the severe phenotype observed in the patient with the 1304N mutation is not produced by alterations in dendritic transport. PMID- 11440197 TI - Transient increase in rab 3A and synaptobrevin immunoreactivity after mild hypoxia in neonatal rats. AB - 1. In the present work we describe the short term effects of mild neonatal hypoxia on the synapse as assessed by the immunoreactivity (IR) of two synaptic proteins: rab 3A and synaptobrevin (VAMP). 2. Using the sensitive methodology of immunoblotting, we measured rab 3A and VAMP-IR in homogenates from the cerebral cortex, hippocampus, and corpus striatum of control (breathing room air) and hypoxiated (breathing 95.5% N2-6.5% O2 for 70 min) 4-day-old rats at 1, 2, and 6 h after the end of the hypoxia. Immunostaining with examination by light microscopy was performed using the synaptic protein-specific antibodies on fixed brain sections from animals belonging to the same litter and submitted to hypoxia. 3. A transient increase of VAMP-IR was observed in the hippocampus and corpus striatum, and for rab 3A in the striatum, 1 h after initiating reoxygenation. At the following time points the values returned to control levels. This effect was less clearly observed in the immunostained sections. 4. Mild hypoxia has an effect on sensitive brain regions, eliciting an increase in the IR of at least two proteins involved in the synaptic vesicle cycle. The transient nature of this effect possibly indicates the activation of endogenous neuroprotective mechanisms. PMID- 11440198 TI - The pharmacodynamic characterization of an antisense oligonucleotide against monoamine oxidase-B (MAO-B) in rat brain striatal tissue. AB - 1. The aim of our work was to pharmacodynamically characterize an antisense oligonucleotide sequence (5'-GCC AAA CTT TTG CAT GAC-3') against MAO-B, using qualitative and quantitative analyses as assessment measures. 2. Qualitative analysis using histochemical staining revealed that intracerebroventricular (ICV) administered antisense (100 picomoles twice daily x 3.5 days) eliminated all visibly detectable histochemical staining for MAO-B throughout the striatum 1, 12, and 24 h after the last antisense treatment. 3. Qualitative analysis using RT PCR of the time course of MAO-B mRNA expression in the rat striatum following ICV administration of the antisense sequence showed that 12-24 h after the last administration there was a dramatic reduction in MAO-B mRNA expression in the striatum. The reverse and scrambled sequences generated no change in MAO-B mRNA at 1 or 24 h after the last treatment. 4. Quantitative analysis using the MAO-B selective substrate 4-dimethylamino-phenethylamine (DMAPEA) showed that the antisense sequence reduced MAO-B activity by more than 40%, which was comparable to a single 2 mg/kg, ip dose of L-deprenyl. 5. Quantitative analysis of neurotransmitter levels 24 h after the last treatment suggested that the antisense sequence did not produce any significant changes in neurotransmitter levels. 6. Potential mechanisms for enhancing the antisense response and the speculated potential of an antisense against MAO-B for studying neurotoxicity, Parkinson's disease, and the aging process are also discussed. PMID- 11440199 TI - Effects of altered cyclophilin A expression on growth and differentiation of human and mouse neuronal cells. AB - 1. Cyclophilin A (CyP-A), a soluble cytoplasmic immunophilin, is known for its involvement in T cell differentiation and proliferation. Although CyP-A has a pivotal role in the immune response, it is most highly concentrated in brain, where its functions are largely unknown. 2. We reported previously that a murine neuroblastoma (NB-P2) cell line can partially differentiate into neurons when treated with cyclosporin A (CyS-A), implicating a role for CyP-A in neuronal differentiation (Hovland et al. [1999]. Neurochem. Int. 3:229-235). 3. The role of CyP-A in regulating neuronal growth and differentiation is not well defined. To investigate this, we first tested the utility of retroviral-mediated gene transfer and expression in human embryonic brain (HEB) and NB-P2 cells. Second, we examined the effects of retroviral-mediated overexpression or antisense mediated reduction of CyP-A in HEB and NB-P2 cells. 4. Our data show that retroviral vectors are efficient for stable gene transfer and expression in both cell lines. Moreover, neither overexpression nor reduction of CyP-A expression in NB-P2 cells altered the growth rate or induced differentiation. More importantly, the up-or down-regulation of CyP-A expression did not affect the magnitude of cAMP-induced NB-P2 differentiation. However, overexpression of CyP-A increased the growth rate of HEB cells. 5. In summary, the utility of retroviral vectors for stable gene expression in human embryonic brain and murine neuroblastoma cells was shown. Furthermore, a novel role for CyP-A in augmenting the proliferation of human embryonic brain cells was demonstrated in vitro. PMID- 11440200 TI - Effect of NMDA-induced lesion of the subfornical organ on the angiotensin II binding sites density and acetylcholinesterase or NADPH-diphorase activities in the lamina terminalis of the rat brain. AB - 1. Neural angiotensinergic circuitry located in the lamina terminalis has been proposed to be involved in blood pressure regulation and fluid homeostasis. 2. ANG II binding sites have been described to be localized throughout the lamina terminalis including the subfornical organ (SFO), the median preoptic nucleus (MnPO), and the organum vasculosum lamina terminalis (OVLT). 3. The present experiment was designed to investigate the ANG II binding sites localization in the lamina terminalis. For this purpose, we have compared the ANG II binding sites, acetylcholinesterase, and NADPH-diaphorase distributions throughout the lamina terminalis. Additionally, we have studied the effect of the preferential lesion of SFO neuronal cell bodies by local injection of NMDA on the ANG II binding sites density in different areas of the lamina terminalis. 4. Male Wistar rats were anesthetized, immobilized in a stereotaxic apparatus, and 500 nl of saline or 250 nmol NMDA was injected into the SFO. 5. Animals were sacrificed 1 week later, the brain was removed, frozen, and sagittal 16 microm slices were cut in a cryostat. Alternate brain slices were incubated with [125I]-Sar1-ANG II for receptor autoradiography or histochemically stained for visualization of acetylcholinesterase and NADPH-diaphorase activities. Binding capacity was determined by computerized quantitative densitometry of autoradiograms. The intensity of histochemical reactions was measured as relative units obtained by computerized densitometry processing of the brain slices stained for either activity. 6. Acetylcholinesterase staining was mainly located in the SFO, with faint staining reaction in other areas of the lamina terminalis. NADPH-diaphorase staining was homogeneously distributed throughout the lamina terminalis. A significant positive correlation was observed between acetylcholinesterase and NADPH-diaphorase stainings in the SFO of control and NMDA-lesioned rats. 7. ANG II binding sites were localized throughout the lamina terminalis. A significant positive correlation was observed between the density of ANG II binding sites and the intensity of acetylcholinesterase or NADPH-diaphorase staining in the SFO of control and NMDA-lesioned rats. 8. The distribution of the NADPH-diaphorase staining was found to closely match the distribution of the ANG II binding sites in the lamina terminalis. 9. Neuronal lesion of the SFO caused significant reductions in the density of ANG II biding sites in the SFO (-68%) and the MnPO ( 48%). No changes were observed either in the OVLT or outside the lamina terminalis in the superior colliculus. 10. The present results indicate the following: first, the presence of high levels of acetylcholinesterase staining in the SFO and of NADPH-diaphorase throughout the lamina terminalis; second, that ANG II binding sites in the SFO and possibly in the MnPO are localized in neuronal cell bodies; third, that SFO lesion did not affect the expression of ANG II binding sites in the OVLT, thus suggesting that these binding sites correspond to different angiotensinergic system: and finally, the existence of a striking correlation between the distribution of the ANG II binding sites and NADPH diaphorase throughout the lamina terminalis, thus suggesting a interrelation between angiotensinergic and nitrergic systems in the lamina terminalis. PMID- 11440201 TI - Rat strain differences in responses of plasma prolactin and PRL mRNA expression after acute amphetamine treatment or restraint stress. AB - 1. The aim of this study was to compare the effects of acute amphetamine (AMPH) treatment and restraint stress on plasma level of prolactin (PRL) and PRL mRNA expression in the adenohypophysis in Sprague-Dawley and Lewis male rats, the latter known to have a deficient hypothalamo-pituitary-adrenal (HPA) axis. 2. Both restraint stress and AMPH treatment (i.p. in a dose of 8 mg/kg of b.w.) were applied 15 or 30 min before termination of the experiment. Plasma PRL and corticosterone (CORT) were determined by radioimmunoassay. PRL mRNA expression was estimated by a dot-blot hybridization. 3. Restraint stress and AMPH treatment induced a significant increase in the CORT plasma level, as an indicator of stress response. Compared to Sprague-Dawley rats, the magnitude of CORT increase after both stimuli was significantly lower in Lewis rats. 4. Although restraint stress significantly increased the PRL plasma levels in both rat strains, AMPH treatment reduced the PRL levels in both rat strains. However, the changes of PRL plasma levels had another pattern in Lewis rats than in Sprague-Dawley rats. Control plasma PRL levels were significantly higher in Lewis rats, and in this rat strain AMPH treatment for 30 min increased the PRL levels as compared to the values obtained after AMPH treatment for 15 min. 5. Expression of PRL mRNA in adenohypophysis by restraint stress and AMPH treatment had a similar pattern. After a 15-min lasting restraint stress, the expression of PRL mRNA was decreased insignificantly in both rat strains. AMPH treatment induced in Sprague-Dawley rats a significant decrease of PRL mRNA after a 15-min interval while after 30 min there was a significant increase. However, in Lewis rats AMPH failed to significantly change PRL mRNA. 6. The results from the present study indicate that the mechanisms mediating the effects of acute restraint stress and acute AMPH treatment differ in PRL response in Sprague-Dawley and Lewis male rat strains. Differences in the observed responses in Lewis rats could be related to the deficient activity of HPA axis in this rat strain. PMID- 11440202 TI - Cat-scratch disease: epidemiology, aetiology and treatment. AB - Cat-scratch disease (CSD) is a clinical syndrome that usually presents as a self limiting lymphadenopathy associated with a cat scratch or bite. Commonly affecting children and young adults, it has a worldwide distribution. In temperate climates, higher rates are reported in the autumn and winter, which can be attributed to the seasonal breeding of the domestic cat. The organism responsible was identified in 1983, having eluded detection for 50 years. Initially, Afipia felis was thought to be the cause; however, subsequent study failed to confirm a link. During the 1990s, it was demonstrated conclusively that Rochalimaea henselae, later reclassified as Bartonella henselae, was the cause of CSD. B. henselae has been isolated from bacteraemic cats, with transmission among cats thought to be via the cat flea. Although other Bartonella species are transmitted by arthropod vectors, it is unlikely that the cat flea is involved directly in human infection, but plays a role in amplifying the reservoir. B. henselae is difficult to culture, and either serology or the polymerase chain reaction are considered to be the best methods of detection. Genetic variation occurs amongst B. henselae strains, perhaps explaining the inconsistency of some diagnostic techniques. A separate serogroup (Marseilles) has been reported in a seronegative patient with CSD, and B. clarridgeiae has the potential to cause the disease. Atypical presentation is seen in up to 25% of cases, and manifests itself as ocular involvement, encephalopathy, granulomatous hepatitis, hepatosplenic infection, endocarditis and osteomyelitis. The majority of CSD cases resolve spontaneously and do not require antibiotic treatment. In complicated CSD, treatment with trimethoprim-sulphamethoxazole, ciprofloxacin or azithromycin is recommended, with gentamicin being reserved for the severely ill patient. PMID- 11440203 TI - Type A gamma-aminobutyric acid (GABAA) receptor subunits and benzodiazepine binding: significance to clinical syndromes and their treatment. AB - Gamma (gamma)-aminobutyric acid (GABA) acting via GABAA receptors is the brain's major inhibitory neurotransmitter system and exerts a crucial role in regulating brain excitability. A number of drugs interact with binding sites on GABAA receptors, and these include benzodiazepines, anticonvulsants, anaesthetics and neurosteroids (e.g. the progesterone metabolite pregnalone). GABAA receptors comprise five subunits (19 are known currently), and are classified into three major groups (alpha, beta and gamma) and several minor ones. The subunit make-up of a receptor, particularly its alpha-subunit content, determines its pharmacological characteristics. Thus, receptors that include an alpha1 subunit have a benzodiazepine (BZ) type I (BZ[I]) pharmacology and bind zolpidem and CL218,872 with high affinity, whilst receptors with alpha2, alpha3 or alpha5 subunits have a BZ type II (BZ[II]) pharmacology and bind these drugs with low affinity. In contrast to receptors that contain alpha4 and alpha6 subunits, which are diazepam-insensitive, both BZ(I) and -(II) bind diazepam and other benzodiazepines. The ligand selectivity of receptor subunits assists in their characterisation. Using immunochemical and ligand-binding techniques, the subunit composition of GABAA receptors has been shown to exhibit a degree of brain regional specificity. GABAA receptors are of great clinical significance in several disorders, including epilepsy, anxiety and alcoholism. In addition to treating epilepsy with drugs that target GABAA and BZ binding sites, epileptic lesions can be localised presurgically using radiolabelled BZ ligands. BZs are used commonly to treat anxiety, and studies suggest that BZ antagonists and inverse agonists (which induce the opposite effect to agonists at receptors) may be useful in alcohol rehabilitation. PMID- 11440204 TI - Kawasaki disease: is it caused by an infectious agent? AB - Kawasaki disease (KD) is an acute systemic febrile illness of unknown aetiology, predominantly affecting children under five years of age. Initially described in 1967 by Tomisaku Kawasaki, it is now the most common cause of acquired heart disease in children in the developed world. Although normally self-limiting, KD is associated with a range of complications, the most important of which is the development of life-threatening coronary artery abnormalities. Here, we examine the evidence supporting the concept that KD is caused by an infectious agent. Various infectious agents--including bacterial, viral and Rickettsial organisms- have been implicated as potential causes, as have certain immunological agents such as bacterial toxin-mediated superantigens, allergens such as anionic detergents and house-dust mites, and some chemicals (including heavy metals). Following extensive research, however, no links between any of these individual agents and the disease have been established irrefutably. Despite this, most of the epidemiological and immunological evidence currently available indicates that the causative agent is most likely to be infectious in nature; and additional evidence highlights the likelihood that development of KD is multifactorial in nature, requiring certain genetic and immunological factors, and possibly a vector. PMID- 11440205 TI - Blastocystis hominis: a common yet neglected human parasite. PMID- 11440206 TI - Presumptive identification and reporting of Streptococcus pneumoniae. PMID- 11440207 TI - Helicobacter pylori: an overview. PMID- 11440208 TI - Urine test for the assessment of smoking status. AB - A simple, quick and inexpensive test for smoking status would be useful in a variety of settings. The non-polar barbituric acid derivative 1,3-dibutyl-2 thiobarbituric acid (DBTB) is described as a novel derivatisation reagent for nicotine and its metabolites in the Konig reaction to assess smoking status. The relative performance of qualitative methods for assessing smoking status using DBTB and the previously employed derivatisation reagent 1,3-diethyl-2 thiobarbituric acid (DETB), as well as quantitatively-based methods for determining 'total nicotine metabolites' (TNMs) using these two reagents, were evaluated against a cotinine-based radioimmunoassay (RIA) as the 'gold standard'. Clinical sensitivity and specificity for all the approaches studied were in excess of 94%. Simple qualitative assessment by eye was superior to quantitatively-based measures of smoking status. Correlation between estimation of nicotine metabolites using DBTB, DETB and RIA were good. The most efficient and convenient method to distinguish between smokers and non-smokers was the simple qualitative method using the more lipophilic reagent DBTB. PMID- 11440209 TI - Ammonia vapour in the mouth as a diagnostic marker for Helicobacter pylori infection: preliminary 'proof of principle' pharmacological investigations. AB - Most current non-invasive tests for Helicobacter pylori depend on the conversion of labelled (13C or 14C) urea to labelled carbon dioxide (13CO2 or 14CO2) and ammonium (NH4+) by the enzyme urease, with the labelled CO2 detected in exhaled air. Despite suggestions going back over a number of years, the alternative possibility of using NH4+ (in the form of gaseous ammonia [NH3]) as the test parameter has received little or no attention. However, this approach is now being explored using a chemiresistive sensor detecting sub-parts per million concentrations of NH3. An in vitro 'glass stomach' (containing various volumes of hydrochloric acid [HCl] and ammonium chloride [NH4Cl]) was used to evaluate the means of increasing 'gastric' pH to that of the NH4+-->NH3 transition that occurs significantly at pH 9.24. This 'stomach' also was used to study mechanisms by which NH3 may be expelled in a pulse (as a surrogate belch), either by the in situ production of CO2 or through an exogenous source. On the basis of the protocols developed, H. pylori-negative subjects were tested before and after ingestion of 10 mg NH4Cl (as a surrogate for bacteria-produced NH4,), and H. pylori-positive subjects were tested without taking urea or NH4Cl. 'Intragastric' pH in the in vitro 'glass stomach' could be increased above pH 9.24 by adding a mixture of 15-30 mL magnesium hydroxide mixture (or the proprietary equivalent) and 50 mL water, and the resulting NH3 expelled by adding 100 mL CO2-saturated cold water (sparkling water). In vivo, NH3 levels in the oral cavity of H. pylori negative subjects were increased after ingestion of 10 mg NH4Cl; however, levels in the oral cavity of a small number of H. pylori-positive subjects were two- to threefold higher after magnesium hydroxide and sparkling water. On the basis of in vitro studies, an in vivo protocol was developed to increase gastric pH above that required for the NH4+-->NH3 transition, and a mechanism established to release the NH3 into the oral cavity. Preliminary in vivo data confirm the chemiresistive sensor is sufficiently sensitive to NH3 to distinguish H. pylori negative subjects who have taken 10 mg NH4Cl from those who have not, and clearly distinguish H. pylori-negative subjects from H. pylori-positive subjects. Ingestion of urea or other labelled tracers is not required, nor is belching; and the sensor takes less than two minutes to reach a maximum response. The data provide good evidence that the chemiresistive detection of NH3 has considerable potential as a rapid, point-of-care diagnostic test for H. pylori infection. PMID- 11440210 TI - C-kit mutation screening in patients with acute myeloid leukaemia: adaptation of a Giemsa-stained bone-marrow smear DNA extraction technique. AB - The scarcity of viable tissue samples for leukaemia research is widely recognised and currently restrictive. Archival bone-marrow smears present a valuable resource that can be exploited easily for mutational analysis. Here, a modified technique to extract DNA is described, and used subsequently for mutation/polymorphism screening of the stem-cell factor receptor proto-oncogene c kit in 23 patients with acute myeloid leukaemia (AML). The selected method was straightforward and used bone-marrow material scraped from periodic acid-Schiff, sudan black B and May-Grunwald/Giemsa-stained preparations, and treated initially with proteinase K prepared in digestion buffer to digest all proteinaceous matter. Following incubation, saturated sodium chloride was added and DNA extracted from the supernatant by phenol/chloroform/isoamyl alcohol treatment. Retrieved DNA was precipitated with ethanol at -20 degrees C overnight, washed with 95% ethanol, air-dried, resuspended using purite water and stored at -20 degrees C prior to use in mutational analysis. The extraction method described was compared with a commercial reagent for combined DNA, RNA and protein isolation using cryopreserved cells from 20 patients with AML. The quality of extracted DNA isolated by the two methods was comparable by polymerase chain reaction (PCR) and single-strand conformation polymorphism (SSCP) techniques. Bone-marrow biopsies are performed regularly on each AML patient to monitor the disease; therefore, an extraction method using this resource could liberate a valuable source of DNA for study (e.g. molecular investigations, including mutation/polymorphism screening etc.). This would allow fresh and programme frozen cells to be reserved for those investigations requiring intact, viable cells. The use of archived bone-marrow smears would permit vast increase in the scope for retrospective testing and large-scale analyses. PMID- 11440211 TI - Human papillomavirus infection in non-neoplastic uterine cervical disease in Hong Kong. AB - The polymerase chain reaction (PCR) was used to detect and identify human papillomavirus (HPV) in 108 cases of formalin-fixed, paraffin-embedded, non neoplastic uterine cervical biopsy tissue retrieved from the surgical pathology archives of the Department of Pathology, Caritas Medical Centre, Hong Kong. After DNA extraction, HPV L1 gene primers were used to detect the presence of HPV, and type-specific primers (to HPV types 6, 11, 16, 18, 31 and 33) were used to identify the specific HPV type on HPV L1-positive cases. PCR amplification of the beta-globin gene was used to ensure the quality of amplifiable DNA extracted. Of 94 cases that yielded sufficient good-quality DNA for PCR analysis, three (one endocervical polyp, one chronic inflammation with erosion, and a normal biopsy) had detectable HPV infection. Two of these had high-risk HPV type 16; the other had an uncommon HPV type. In view of the low incidence of HPV found in these patients, large-scale population screening of clinical samples using PCR to detect the presence of HPV and identify high-risk asymptomatic patients would not be cost-effective. PMID- 11440212 TI - Effect of cell washing on erythrocyte sodium concentration in human hypertensive subjects. PMID- 11440213 TI - Dietary probiotic supplementation to enhance cellular immunity in the elderly. PMID- 11440214 TI - Seroprevalence of Helicobacter pylori infection in patients with gastritis and peptic ulcer in western Nigeria. PMID- 11440215 TI - Management of infections caused by respiratory syncytial virus. AB - This is a consensus document compiled by the Medical Products Agency in Sweden and the Swedish Reference Group for Antiviral Therapy on management of respiratory syncytial virus (RSV) infections. Prophylaxis against RSV infections using palivizumab, a commercially available humanized monoclonal IgG1 antibody preparation, is recommended for children <2 y of age with chronic respiratory diseases requiring continuous treatment (oxygen and/or inhalations and/or steroids) during the previous 6 months and children <6 months old who were born before gestational week 26. Ribavirin inhalation treatment may be considered in high-risk infants with clinical symptoms indicating a serious course of an RSV infection. Treatment with ribavirin in combination with intravenous polyclonal immunoglobulin should be considered in patients who have received an allogenic stem cell transplantation or organ transplantation with >1 episode of rejection treatment and who have mild or moderate RSV pneumonia. Evidence-based documentation for treatment of other groups of patients is lacking. PMID- 11440216 TI - The global control of tuberculosis: what are the prospects? AB - Tuberculosis (TB) still imposes a huge burden of ill health, premature death and emotional suffering on the developing world. Over the past 30 y it has been greatly neglected by those concerned about international public health and there are now nearly 8 million new cases annually and 1.86 million deaths. An epidemic of HIV-associated TB is now affecting Africa and threatening parts of Asia. Multidrug-resistant TB has emerged as a huge threat in Russia and its former satellites. However, with the advent of the directly observed treatment, short course (DOTS) strategy in 1995, high-burden countries have started to seriously address the problem. Recent political commitment on the part of the rich nations, together with significant increases in funding from private foundations and great scientific advances in our understanding of Mycobacterium tuberculosis, give rise to cautious optimism that TB will be controlled during this century. PMID- 11440217 TI - Antibiotic susceptibility patterns of community- and hospital-acquired Staphylococcus aureus and Escherichia coli in Estonia. AB - This study compares the susceptibility patterns of Staphylococcus aureus and Escherichia coli isolated from patients with hospital-acquired and outpatient infections. A total of 902 isolates of S. aureus and 1,114 of E. coli were collected in five different Estonian medical centers between January 1997 and November 1997. Strains were grouped into two different categories, depending on whether they had been obtained from inpatients or outpatients. Compared to S. aureus strains isolated from inpatients, the strains from outpatients were significantly more resistant to erythromycin (25.3% vs. 17.9%), tetracycline (33.5% vs. 22.4%) and trimethoprim-sulfamethoxazole (13.9% vs. 7.9%). The overall prevalence of oxacillin-resistant S. aureus was 10.4%, with no significant differences noted between isolates recovered from inpatients and outpatients. In the case of E. coli, significantly more isolates from inpatients (42.8%) than from outpatients (34.4%) were ampicillin-resistant. Inpatient isolates of E. coli were also more resistant to cefotaxime (9.3%) and nitrofurantoin (11.2%) than outpatient strains (0% and 3.1%, respectively). Analysis showed remarkable co resistance among both inpatient and outpatient strains of S. aureus and E. coli. Multiple resistant S. aureus and E. coli strains represented 15.1% and 17.3%, respectively of the organisms examined in this study. With respect to E. coli, significantly more multiresistant isolates were found in inpatient than outpatient isolates (20.4% vs. 8.9%). Our results indicate that the distinction between community-acquired and hospital infections due to S. aureus and E. coli may not be valid in Estonia. PMID- 11440218 TI - Fewer bacterial relapses after oral treatment with norfloxacin than with ceftibuten in acute pyelonephritis initially treated with intravenous cefuroxime. AB - This double-blind, multicentre study was performed at nine centres on a total of 171 patients who presented with fever (> 38.5 degrees C) and signs of acute pyelonephritis. All were initially treated with intravenous cefuroxime. After 2-3 d, when the fever had subsided and urinary culture had revealed growth of Gram negative bacteria ( > 10(7) colony-forming units per litre), treatment was changed to oral administration of ceftibuten 200 mg b.i.d. or norfloxacin 400 mg b.i.d. for 10 d. The patients were followed for signs of bacterial or clinical relapse 7-14 d after the end of treatment. The initial clinical and bacteriological cure was excellent in both groups, but there were significantly fewer bacterial relapses after oral treatment with norfloxacin than with ceftibuten in acute febrile pyelonephritis initially treated with intravenous cefuroxime. The causal strain was eradicated in 75% of patients (73% of males, 76% of females) in the ceftibuten group and in 89% of patients (94% of males, 85% of females) in the norfloxacin group. The relative frequency of eradication was 0.84 (p < 0.05; 95%, confidence interval 0.74-0.97). Adverse events were reported by 47% of the patients in the ceftibuten group and by 38% in the norfloxacin group. This difference was not significant, but diarrhoea or loose stools occurred more frequently in the ceftibuten group. PMID- 11440219 TI - Antibacterial susceptibility of intestinal lactobacilli of healthy children. AB - We investigated the antibacterial susceptibility of intestinal lactobacilli of Estonian and Swedish children aged 1-2 y. Sixty isolates (10 species) of lactobacilli (29 Estonian and 31 Swedish strains) were tested against ampicillin, cefuroxime, cefoxitin, gentamicin, ciprofloxacin, tetracycline, vancomycin, metronidazole and erythromycin. We observed that intestinal lactobacilli do not display uniform susceptibility to antibiotics. None of the tested lactobacilli was resistant to ampicillin, gentamicin and erythromycin. Single strains were resistant to cefuroxime and tetracycline, about half of the strains to cefoxitin and ciprofloxacin and 73% of the strains to vancomycin. All studied strains were resistant to metronidazole. Most of the strains investigated were resistant to two or three antibiotics out of nine. Some differences in susceptibility were noted between strains belonging to different fermentation types. No differences in susceptibility were found between Estonian and Swedish isolates. Metronidazole, cefoxitin, vancomycin and ciprofloxacin seem to be safer for gastrointestinal lactoflora than other tested antibiotics in both countries. PMID- 11440220 TI - Circulating soluble vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 in immunocompetent and renal transplant patients: correlation with cytomegalovirus disease and renal function. AB - The plasma levels of the soluble adhesion molecules, soluble vascular cell adhesion molecule-1 (sVCAM-1) and intercellular adhesion molecule-1 (sICAM-1), were measured before and after transplantation in 26 renal transplant recipients, and in 173 longitudinally collected samples in 17 of the patients. The patients were carefully monitored for the presence of cytomegalovirus (CMV) infection and rejection. Forty healthy blood donors and 12 otherwise healthy subjects with symptomatic primary CMV infections served as controls. During CMV disease, plasma levels of sVCAM-1 and sICAM-1 were elevated in both renal transplant patients and otherwise healthy subjects with CMV disease. The sVCAM-1 levels were strongly elevated before transplantation in renal transplant recipients and correlated with creatinine levels. Increased sVCAM-1 levels were also registered during rejection episodes. CMV disease, per se, is associated with markedly increased levels of sVCAM-1 and sICAM-1. There is also a correlation of sVCAM-1 levels with serum creatinine levels. Thus, the presence of CMV infection and renal function are factors that must be considered in further studies of soluble adhesion molecules. PMID- 11440221 TI - An evaluation of the diagnostic value of clinical and radiological manifestations in patients attending the addis ababa tuberculosis centre. AB - Our investigation is based on 1,000 consecutive patients attending the routine outpatient services of the Addis Ababa Tuberculosis Demonstration and Training Centre (ATBDTC) in Addis Ababa during the period November 1996-March 1997. By using sputum microscopy for acid-fast bacilli (AFB), chest radiography (CXR) and clinical assessment these patients had been diagnosed as having either AFB positive pulmonary tuberculosis (PTB) (n = 139), AFB-negative PTB (n = 61) or non tuberculosis (non-TB) (n = 800). These three diagnostic groups were subsequently re-assessed by us with regard to selected demographic and clinical parameters, including CXR, in order to identify and weight markers significantly linked to proven PTB. The sum of the individual weights provided diagnostic scores (DS); the average DS for AFB-positive patients was 653.5 +/- 174 and that for non-TB patients was 219.1 +/- 138.7. The calculated cut-off value between these two groups was 444. Ten (7.2%) AFB-positive PTB patents had a DS below the calculated cut-off, while 46 (5.8%) of the 800 non-TB cases had diagnostic scores exceeding this value. Our DS system achieved 92.8% sensitivity and 94.3% specificity, with positive and negative predictive values of 73.7% and 98.7%, respectively. When plotting the individual DS values of the 61 AFB-negative TB patients, 24 (39.3%) fell below the cut-off. It is most likely that these patients did not have PTB. We conclude that there is a need for improved and standardized diagnostic approaches for TB-suspected patients who depend upon clinical and CXR criteria for correct diagnosis. Our score system may be helpful in this context. PMID- 11440222 TI - Mycobacterial cervical lymphadenitis in children: clinical and laboratory factors of importance for differential diagnosis. AB - The role of epidemiological and clinical features for the early differential diagnosis between Mycobacterium tuberculosis (MTB) complex and non-tuberculous mycobacterial (NTM) cervical lymphadenitis in children was examined in this study. From 1982 to 1997, 24 MTB complex cases and 26 NTM cases were diagnosed, of which 75% and 25%, respectively, presented during the first half of the study period. Epidemiological and lymph-node features and anatomical areas were not helpful in the differential diagnosis between the 2 groups. Fulfilment of 2 out of 3 criteria (positive tuberculin skin test reaction, abnormal chest radiograph, contact with a person with infectious tuberculosis) was associated with 92% sensitivity for the diagnosis of MTB lymphadenitis. 37.5% of the MTB cases and 88.5% of the NTM cases were culture-confirmed; all inconclusive cultures concerned patients with spontaneous drainage and fistula. Surgical intervention was required in 67% of the MTB cases. All NTM cases were managed by surgery alone. Fistulae or cheloids occurred in all patients in whom incision and drainage were applied instead of total excision. Excellent aesthetic results were achieved in patients who presented within 1 month following the onset of lymphadenitis. The spectrum of mycobacterial cervical lymphadenitis in children in Greece has changed during the 1990s. Early and prompt treatment contributes to the diagnosis and response. PMID- 11440223 TI - Prospective study of fungaemia in a single cancer institution over a 10-y period: aetiology, risk factors, consumption of antifungals and outcome in 140 patients. AB - Over a 10-y period (1989-99) we prospectively evaluated all patients with fungaemia among 16,555 admissions (21,004 blood cultures) at a national cancer referral institution in the Slovak Republic. A prospective protocol was completed on 140 patients with fungaemia, which was then analysed in terms of aetiology, clinical characteristics, potential risk factors and outcome. The most frequently isolated organism was C. albicans, in 75 patients (52.9%), followed by non albicans Candida spp. in 45 patients (32.1%). Non-Candida spp. yeasts represented 16 episodes in 16 patients (11.4%). Moulds caused 4 episodes in 4 patients (3.6% of all fungaemias) and all were caused by Fusarium spp. Mucositis (p = 0.025), > or = 3 positive blood cultures (p = 0.02), acute leukaemia (p = 0.00001), neutropenia (p = 0.0015), quinolone prophylaxis (p < 0.000005) and breakthrough fungaemia (p = 0.004) during prophylaxis with fluconazole (p = 0.03) and itraconazole (p = 0.005) were significantly more associated with non-Candida than C. albicans spp. Furthermore, attributable mortality was higher in the subgroup of non-Candida than C. albicans spp. (50.0 vs. 18.7%, p < 0.02). The only independent risk factor for inferior outcome was antifungal therapy of < 10 d duration (odds ratio 2.1, 95% confidence interval, p < 0.001). Aetiology, neutropenia and mucositis were not independent risk factors for higher mortality in multivariate analysis; however, they were risk factors for inferior outcome in univariate analysis (p < 0.05-0.005). PMID- 11440224 TI - Successful treatment of vancomycin-resistant Enterococcus faecium meningitis with linezolid: case report and literature review. AB - Vancomycin-resistant enterococci (VRE) are a rare cause of meningitis, occurring primarily in patients who have undergone neurosurgical procedures. We describe the first reported case of VRE meningitis successfully treated with linezolid. A 56-y-old female with subarachnoid hemorrhage underwent ventriculostomy and embolization of cerebral aneurysms. Her postoperative course was complicated by multiple infections needing repeated antibiotic courses, culminating in the development of VRE meningitis. She was treated with 600 mg of i.v. linezolid (MIC < 0.75 microg/ml) every 12 h for 6 weeks. After the fourth dose, peak and trough linezolid concentrations were 11.45 and 0.14 microg/ml in serum and 3.19 and 2.39 microg/ml in cerebral spinal fluid (CSF). On Day 19 of linezolid therapy, serum and CSF trough concentrations were 1.53 and 2.98 microg/ml, respectively. Linezolid achieved sufficient CSF concentrations to bring about clinical and bacteriological cure. We conclude that i.v. linezolid may be a useful option for treating VRE meningitis. We also present findings of a literature review, which identified 11 cases of VRE meningitis treated with other pharmacologic agents with mixed success. PMID- 11440225 TI - Propionibacterium acnes endocarditis in a native valve complicated by intraventricular abscess: a case report and review. AB - Propionibacterium acnes is a constituent of the normal skin flora. It has been described as causing infection on prosthetic valves but very rarely on native valves. We describe a case of aggressive P. acnes endocarditis in a healthy 36-y old man which infected a native aortic valve and was complicated by an aortic root abscess and review the literature. PMID- 11440226 TI - Polymicrobial polyarticular septic arthritis: a rare clinical entity. AB - Polymicrobial polyarticular septic arthritis is a rare clinical entity, with only a few cases having been reported to date. We report a case due to Streptococcus pyogenes and Staphylococcus aureus in an IVDU, complicated by fatal streptococcal toxic-shock syndrome, and review the current literature. We conclude that whenever polymicrobial polyarticular septic arthritis is diagnosed, a high index of suspicion should be maintained for the detection of locally destructive infectious processes as well as systemic complications, and that a high mortality rate should be expected. PMID- 11440227 TI - Rapid laboratory diagnosis of ulceroglandular tularemia with polymerase chain reaction. AB - Tularemia is a zoonotic disease which, in Scandinavia, is usually acquired through a mosquito bite. As the infecting organism, Francisella tularensis, is highly virulent the culturing of F. tularensis has generally been avoided. PCR offers a safe way to rapidly confirm diagnosis of tularemia. The case of a 9-y old boy with ulceroglandular tularemia is presented. The diagnosis was made rapidly with DNA amplification from a pus specimen. The efficacy of ciprofloxacin treatment of tularemia in children is also discussed. PMID- 11440228 TI - Isolation of Haemophilus parahaemolyticus in a patient with cryptogenic brain abscess. AB - Haemophilus parahaemolyticus rarely causes disease in humans. We present the case of a 68-y-old man who suffered from a brain abscess over the right-side thalamus and midbrain. Stereotactic aspirates grew H. parahaemolyticus. No portal of entry for the microorganism was identified in this case. The patient made a good recovery after treatment. PMID- 11440229 TI - Sternal tuberculosis after coronary artery bypass graft surgery. AB - We report a case of sternal tuberculosis following sternotomy, which was performed during coronary artery bypass graft surgery. Although pre-operative evaluation revealed signs of asymptomatic tuberculosis of the lung, isoniazid chemoprophylaxis was not instituted, and the patient developed active tuberculosis in both the lung and sternum 5 y later. PMID- 11440230 TI - Simultaneous occurrence of 2 HIV-related immunereconstitution diseases after initiation of highly active antiretroviral therapy. AB - The case of a 44-y-old woman with HIV infection and cytomegalovirus retinitis in whom antiretroviral therapy (HAART) revealed pulmonary cryptococcosis is presented. Pulmonary cryptococcosis occurred simultaneously with immune recovery uveitis after starting HAART, showing that complex clinical pictures may arise from immunreconstitution diseases. PMID- 11440231 TI - Failure of pre- and postexposure rabies vaccinations in a child infected with HIV. AB - We report the case of a 6-y-old HIV-infected girl with severe immune deficiency who failed to respond to intramuscular pre-exposure rabies vaccination using human diploid cell rabies vaccine on days 0, 7 and 28. She also failed to respond to an intradermal postexposure rabies regimen using purified verocell rabies vaccine at 4 sites on days 0, 3 and 7 and at 2 sites on days 30 and 90 (double the usual regimen). Sequentially monitored rabies neutralizing antibody titers were below the WHO minimum acceptable level (> 0.15 IU/ml) in all specimens. Rabies prevention in HIV-infected persons with severe immune suppression requires further study. PMID- 11440232 TI - Acute, recurrent fosfomycin-induced liver toxicity in an adult patient with cystic fibrosis. AB - We report a very unusual adverse effect--fosfomycin-induced repeat liver toxicity -in a female adult with cystic fibrosis (CF). PMID- 11440233 TI - Capsular polysaccharide types of Streptococcus pneumoniae isolated from maxillary sinus effusion. AB - Pneumococcal capsular polysaccharide vaccines may be a novel way of treating patients with recurrent sinusitis. To evaluate the pneumococcal capsular polysaccharide types of strains causing sinusitis, 55 pneumococcal strains isolated from maxillary sinus effusion were analyzed. The most frequently occurring types were 3 (13%), 19F (11%), 6B (9%) and 23F (9%). PMID- 11440234 TI - Mortality of enterococcal bacteremia: are inappropriately treated cases associated with higher mortality? PMID- 11440235 TI - Comments on PCR-RFLP as an original technique to detect point mutations in the Pneumocystis carinii DHPS gene. PMID- 11440236 TI - Pacemaker endocarditis due to Yersinia enterocolitica. PMID- 11440237 TI - Varicella-zoster virus meningitis and cerebrospinal fluid HIV RNA. PMID- 11440238 TI - Anaerobic benzene degradation. AB - Although many studies have indicated that benzene persists under anaerobic conditions in petroleum-contaminated environments, it has recently been documented that benzene can be anaerobically oxidized with most commonly considered electron acceptors for anaerobic respiration. These include: Fe(III), sulfate, nitrate, and possibly humic substances. Benzene can also be converted to methane and carbon dioxide under methanogenic conditions. There is evidence that benzene can be degraded under in situ conditions in petroleum-contaminated aquifers in which either Fe(III) reduction or methane production is the predominant terminal electron-accepting process. Furthermore, evidence from laboratory studies suggests that benzene may be anaerobically degraded in petroleum-contaminated marine sediments under sulfate-reducing conditions. Laboratory studies have suggested that within the Fe(III) reduction zone of petroleum-contaminated aquifers, benzene degradation can be stimulated with the addition of synthetic chelators which make Fe(III) more available for microbial reduction. The addition of humic substances and other compounds that contain quinone moieties can also stimulate anaerobic benzene degradation in laboratory incubations of Fe(III)-reducing aquifer sediments by providing an electron shuttle between Fe(III)-reducing microorganisms and insoluble Fe(III) oxides. Anaerobic benzene degradation in aquifer sediments can be stimulated with the addition of sulfate, but in some instances an inoculum of benzene-oxidizing, sulfate-reducing microorganisms must also be added. In a field trial, sulfate addition to the methanogenic zone of a petroleum-contaminated aquifer stimulated the growth and activity of sulfate-reducing microorganisms and enhanced benzene removal. Molecular phylogenetic studies have provided indications of what microorganisms might be involved in anaerobic benzene degradation in aquifers. The major factor limiting further understanding of anaerobic benzene degradation is the lack of a pure culture of an organism capable of anaerobic benzene degradation. PMID- 11440239 TI - Evidence for aromatic ring reduction in the biodegradation pathway of carboxylated naphthalene by a sulfate reducing consortium. AB - Naphthalene was used as a model compound in order to study the anaerobic pathway of polycyclic aromatic hydrocarbon degradation. Previously we had determined that carboxylation is an initial step for anaerobic metabolism of naphthalene, but no other intermediate metabolites were identified (Zhang & Young 1997). In the present study we further elucidate the pathway with the identification of six novel naphthalene metabolites detected when cultures were fed naphthalene in the presence of its analog 1 -fluoronaphthalene. Results from cultures supplemented with either deuterated naphthalene or non-deuterated naphthalene plus [13C]bicarbonate confirm that the metabolites originated from naphthalene. Three of these metabolites were identified by comparison with the following standards: 2-naphthoic acid (2-NA), 5,6,7,8-tetrahydro-2-naphthoic acid, and decahydro-2 naphthoic acid. The presence of 5,6,7,8-tetrahydro-2-NA as a metabolite of naphthalene degradation indicates that the first reduction reaction occurs at the unsubstituted ring, rather than the carboxylated ring. The overall results suggest that after the initial carboxylation of naphthalene, 2-NA is sequentially reduced to decahydro-2-naphthoic acid through 5 hydrogenation reactions, each of which eliminated one double bond. Incorporation of deuterium atoms from D2O into 5,6,7,8-tetrahydro-2-naphthoic acid suggests that water is the proton source for hydrogenation. PMID- 11440240 TI - Metabolic indicators for detecting in situ anaerobic alkylbenzene degradation. AB - Monitoring programs for intrinsic bioremediation of fuel hydrocarbons require indicators that can convincingly demonstrate in situ metabolism. In this evaluation of potential indicators of in situ anaerobic alkylbenzene metabolism, laboratory and field data are reviewed for two classes of aromatic acids: (i) benzylsuccinate, E-phenylitaconate, and their methyl homologs, and (ii) benzoate, and methyl-, dimethyl-, and trimethylbenzoates. The review includes previously unpublished field data from a hydrocarbon-contaminated site in Fallon (Nevada), at which both classes of metabolites were detected in groundwater. The two classes of compounds were evaluated with respect to specificity (i.e., unique biochemical relationship to a specific alkylbenzene), stability, and generation as degradation intermediates versus dead-end products; recent developments in the biochemistry of anaerobic toluene and xylene degradation were incorporated in this evaluation. In general, benzylsuccinates/E-phenylitaconates are superior to benzoates in terms of their very high specificity to their parent hydrocarbons and their lack of commercial and industrial sources. They are also uniquely indicative of anaerobic conditions. All of the benzoates, benzylsuccinates, and E phenylitaconates are relatively stable chemically and (with the exception of benzoate) biologically under anaerobic conditions, based on the limited data available. Although benzoate, benzylsuccinate, and E-phenylitaconate are intermediates of anaerobic toluene mineralization to carbon dioxide, their methyl homologs can be either mineralization intermediates or cometabolic dead-end products of alkylbenzenes, depending on the bacteria involved. Benzoates are far more commonly reported in field studies of hydrocarbon-contaminated aquifers than are benzylsuccinates and E-phenylitaconates, although it is not clear whether this is an accurate representation of the relative occurrence of these metabolites at contaminated sites, or whether it instead reflects the limited range of target analytes used in most field studies to date. PMID- 11440241 TI - Microbial consortia involved in the anaerobic degradation of hydrocarbons. AB - In this review, we examine the energetics of well-characterized biodegradation pathways and explore the possibilities for these to support growth of multiple organisms interacting in consortia. The relevant phenotypic and/or phylogenetic characteristics of isolates and consortia mediating hydrocarbon degradation coupled with different terminal electron-accepting processes (TEAP) are also reviewed. While the information on metabolic pathways has been gained from the analysis of individual isolates, the energetic framework presented here demonstrates that microbial consortia could be readily postulated for hydrocarbon degradation coupled to any TEAP. Several specialized reactions occur within these pathways, and the organisms mediating these are likely to play a key role in defining the hydrocarbon degradation characteristics of the community under a given TEAP. Comparing these processes within and between TEAPs reveals biological unity in that divergent phylotypes display similar degradation mechanisms and biological diversity in that hydrocarbon-degraders closely related as phylotypes differ in the type and variety of hydrocarbon degradation pathways they possess. Analysis of microcosms and of field samples suggests that we have only begun to reveal the diversity of organisms mediating anaerobic hydrocarbon degradation. Advancements in the understanding of how hydrocarbon-degrading communities function will be significantly affected by the extent to which organisms mediating specialized reactions can be identified, and tools developed to allow their study in situ. PMID- 11440242 TI - Enhanced anaerobic bioremediation of groundwater contaminated by fuel hydrocarbons at Seal Beach, California. AB - Enhanced anaerobic biodegradation of groundwater contaminated by fuel hydrocarbons has been evaluated at a field experiment conducted at the Naval Weapons Station, Seal Beach, California. This experiment included the establishment of three different remediation zones in situ: one zone was augmented with sulfate, one was augmented with sulfate and nitrate, and the third was unaugmented. This enables a comparison of hydrocarbon biodegradation under sulfate-reducing, sequential denitrifying/sulfate-reducing, and methanogenic conditions, respectively. In general, the results from the field experiment are: (1) Certain fuel hydrocarbons were removed preferentially over others, but the order of preference is dependent upon the geochemical conditions; and (2) In the zones that were augmented with sulfate and/or nitrate, the added electron acceptors were consumed quickly, indicating that enhancement via electron acceptor injection accelerates the biodegradation process. More specifically, in the sulfate-reducing zone, sulfate was utilized with an apparent first-order rate coefficient of approximately 0.1 day(-1). In the combined denitrifying/sulfate reducing zone, nitrate was utilized preferentially over sulfate, with an apparent first-order rate coefficient of 0.1-0.6 day(-1). However, the data suggest that slow sulfate utilization does occur in the presence of nitrate, i.e., the two processes are not strictly sequential. With regard to the aromatic BTEX hydrocarbons, toluene was preferentially removed under intrinsic conditions; biodegradation of benzene was slow if it occurred at all; augmentation with sulfate preferentially stimulated biodegradation of o-xylene; and ethylbenzene appeared recalcitrant under sulfate-reducing conditions but readily degradable under denitrifying conditions. PMID- 11440243 TI - Aerobic MTBE biodegradation: an examination of past studies, current challenges and future research directions. AB - With the current practice of amending gasoline with up to 15% by volume MTBE, the contamination of groundwater by MTBE has become widespread. As a result, the bioremediation of MTBE-impacted aquifers has become an active area of research. A review of the current literature on the aerobic biodegradation of MTBE reveals that a number of cultures from diverse environments can either partially degrade or completely mineralize MTBE. MTBE is either utilized as a sole carbon and energy source or is degraded cometabolically by cultures grown on alkanes. Reported degradation rates range from 0.3 to 50 mg MTBE/g cells/h while growth rates (0.01-0.05 g MTBE/g cells/d) and cellular yields (0.1-0.2 g cells/g MTBE) are generally low. Studies on the mechanisms of MTBE degradation indicate that a monooxygenase enzyme cleaves the ether bond yielding tert-butyl alcohol (TBA) and formaldehyde as the dominant detectable intermediates. TBA is further degraded to 2-methyl-2-hydroxy-1-propanol, 2-hydroxyisobutyric acid, 2-propanol, acetone, hydroxyacteone and eventually, carbon dioxide. The majority of these intermediates are also common to mammalian MTBE metabolism. Laboratory studies on the degradation of MTBE in the presence of gasoline aromatics reveal that while degradation rates of other gasoline components are generally not inhibited by MTBE, MTBE degradation could be inhibited in the presence of more easily biodegradable compounds. Controlled field studies are clearly needed to elucidate MTBE degradation potential in co-contaminant plumes. Based on the reviewed studies, it is likely that a bioremediation strategy involving direct metabolism, cometabolism, bioaugmentation, or some combination thereof, could be applied as a feasible and cost-effective treatment method for MTBE contamination. PMID- 11440244 TI - Bioremediation of MTBE: a review from a practical perspective. AB - The addition of methyl tert-butyl ether (MTBE) to gasoline has resulted in public uncertainty regarding the continued reliance on biological processes for gasoline remediation. Despite this concern, researchers have shown that MTBE can be effectively degraded in the laboratory under aerobic conditions using pure and mixed cultures with half-lives ranging from 0.04 to 29 days. Ex-situ aerobic fixed-film and aerobic suspended growth bioreactor studies have demonstrated decreases in MTBE concentrations of 83% and 96% with hydraulic residence times of 0.3 hrs and 3 days, respectively. In microcosm and field studies, aerobic biodegradation half-lives range from 2 to 693 days. These half-lives have been shown to decrease with increasing dissolved oxygen concentrations and, in some cases, with the addition of exogenous MTBE-degraders. MTBE concentrations have also been observed to decrease under anaerobic conditions; however, these rates are not as well defined. Several detailed field case studies describing the use of ex-situ reactors, natural attenuation, and bioaugmentation are presented in this paper and demonstrate the potential for successful remediation of MTBE contaminated aquifers. In conclusion, a substantial amount of literature is available which demonstrates that the in-situ biodegradation of MTBE is contingent on achieving aerobic conditions in the contaminated aquifer. PMID- 11440245 TI - Metabolism of alkylbenzenes, alkanes, and other hydrocarbons in anaerobic bacteria. AB - Aromatic and aliphatic hydrocarbons are the main constituents of petroleum and its refined products. Whereas degradation of hydrocarbons by oxygen-respiring microorganisms has been known for about a century, utilization of hydrocarbons under anoxic conditions has been investigated only during the past decade. Diverse strains of anaerobic bacteria have been isolated that degrade toluene anaerobically, using nitrate, iron(III), or sulfate as electron acceptors. Also, other alkylbenzenes such as m-xylene or ethylbenzene are utilized by a number of strains. The capacity for anaerobic utilization of alkylbenzenes has been observed in members of the alpha-, beta-, gamma- and delta-subclasses of the Proteobacteria. Furthermore, denitrifying bacteria and sulfate-reducing bacteria with the capacity for anaerobic alkane degradation have been isolated, which are members of the beta- and delta-subclass, respectively. The mechanism of the activation of hydrocarbons as apolar molecules in the absence of oxygen is of particular interest. The biochemistry of anaerobic toluene degradation has been studied in detail. Toluene is activated by addition to fumarate to yield benzylsuccinate, which is then further metabolized via benzoyl-CoA. The toluene activating enzyme presents a novel type of glycine radical protein. Another principle of anaerobic alkylbenzene activation has been observed in the anaerobic degradation of ethylbenzene. Ethylbenzene in denitrifying bacteria is dehydrogenated to 1-phenylethanol and further to acetophenone; the latter is also metabolized to benzoyl-CoA. Naphthalene is presumably activated under anoxic conditions by a carboxylation reaction. Investigations into the pathway of anaerobic alkane degradation are only at the beginning. The saturated hydrocarbons are most likely activated by addition of a carbon compound rather than by desaturation and hydration, as speculated about in some early studies. An anaerobic oxidation of methane with sulfate as electron acceptor has been documented in aquatic sediments. The process is assumed to involve a reversal of methanogenesis catalyzed by Archaea, and scavenge of an electron-carrying metabolite by sulfate-reducing bacteria. Among unsaturated non-aromatic hydrocarbons, anaerobic bacterial degradation has been demonstrated and investigated with n-alkenes, alkenoic terpenes and the alkyne, acetylene. PMID- 11440246 TI - The "sizes" of mutations fixed in phenotypic evolution: a response to Clarke and Arthur. PMID- 11440247 TI - Why imperfect steps in the right direction attract criticism. PMID- 11440248 TI - Additional hox clusters in the zebrafish: divergent expression patterns belie equivalent activities of duplicate hoxB5 genes. AB - The evolution of metazoan body plans has involved changes to the Hox genes, which are involved in patterning the body axis and display striking evolutionary conservation of structure and expression. Invertebrates contain a single Hox cluster whereas tetrapods possess four clusters. The zebrafish has seven unlinked hox clusters, a finding that is difficult to reconcile with the notion that genomic complexity, reflected by Hox cluster number, and morphological complexity are causally linked, as the body plan of the zebrafish is not obviously more complex than that of the mouse or human. Why have the additional hox genes in zebrafish been conserved? To address the role of these additional zebrafish hox genes, we have examined the duplicate hoxB5 genes, hoxB5a, and hoxB5b. Conservation of gene duplicates can occur when one gene acquires a new function (neofunctionalization), or when the ancestral function is divided between the two duplicates (subfunctionalization). hoxB5a and hoxB5b are expressed in distinct domains, and their combined expression domain is strikingly similar to that of single Hoxb5 genes in other species. The biochemical functions encoded by the two genes were studied by overexpression, which resulted in identical developmental defects in the anterior hindbrain and cranial neural crest, suggesting strongly that hoxB5a and hoxB5b have equivalent biochemical properties with respect to early development. From these studies, we conclude that conservation of hoxB5a and hoxB5b is likely the result of division of the ancestral Hoxb5 function between the two genes, without significant changes in biochemical activity. These results suggest a resolution to the conundrum of the extra hox genes and clusters in the zebrafish, since if any of the additional hox genes in the zebrafish are similarly subfunctionalized, they are unlikely to supply novel genetic functions. Thus, the morphological complexity potentially conferred by the majority of additional zebrafish hox clusters may not be substantially greater than that conferred by the four tetrapod clusters. PMID- 11440249 TI - Hox gene duplication and deployment in the annelid leech Helobdella. AB - The segmented leeches are members of the phylum Annelida within the Lophotrochozoa. Here, we describe the isolation of a new Hox gene, Lox18, in the leech Helobdella triserialis. Phylogenetic analysis indicates that Lox18 is a Deformed (Dfd) ortholog. H. triserialis has at least two Dfd orthologs, Lox18 and the previously described Lox6 (Kourakis et al. 1997; Wong and Macagno 1998), indicating that these genes duplicated after the last common ancestor of annelids and arthropods. Although the temporal appearance of Lox18 message is similar to that of Lox6, the spatial pattern is different. Lox18 does not have a sharply defined anterior border of expression in the second neuromere of the subesophageal ganglion of the central nervous system (CNS) as does Lox6, but is expressed uniformly in a small subset of cells in the longitudinal connectives and lateral roots in every segment of the CNS along the entire anterior-posterior (AP) axis. Even though Lox18 shares greater sequence similarity within the homeodomain and flanking regions to Drosophila Dfd than to the previously isolated Lox6, its expression pattern suggests that its function has diverged from the ancestral Hox function. Previous sampling has indicated that the last common ancestor of protostomes and deuterostomes had as many as 10 clustered Hox genes representing distinct paralogy groups (Irvine et al. 1997; de Rosa et al. 1999); leech Hox genes may have undergone subsequent and independent cluster or genome-wide duplication. These results point to the need for total genome level understanding for key members of the Lophotrochozoa. PMID- 11440250 TI - Neurogenesis in the crustacean ventral nerve cord: homology of neuronal stem cells in Malacostraca and Branchiopoda? AB - In Insecta and malacostracan Crustacea, neurons in the ventral ganglia are generated by the unequal division of neuronal stem cells, the neuroblasts (Nbs), which are arranged in a stereotyped, grid-like pattern. In malacostracans, however, Nbs originate from ectoteloblasts by an invariant lineage, whereas Nbs in insects differentiate without a defined lineage by cell-to-cell interactions within the neuroectoderm. As the ventral ganglia in entomostracan crustaceans were thought to be generated by a general inward proliferation of ectodermal cells, the question arose as to whether neuroblasts in Euarthropoda represent a homologous type of stem cell. In the current project, neurogenesis in metanauplii of the entomostracan crustaceans Triops cancriformis Fabricius, 1780 (Branchiopoda, Phyllopoda) and Artemia salina Linne, 1758 (Branchiopoda, Anostraca) was examined by in vivo incorporation of the mitosis marker bromodeoxyuridine (BrdU) and compared to stem cell proliferation in embryos of the malacostracan Palaemonetes argentinus Nobili, 1901 (Eucarida, Decapoda). The developmental expression of synaptic proteins (synapsins) was studied immunohistochemically. Results indicate that in the ventral neurogenic zone of Branchiopoda, neuronal stem cells with cellular characteristics of malacostracan neuroblasts are present. However, a pattern similar to the lineage-dependent, grid-like arrangement of the malacostracan neuroblasts was not found. Therefore, the homology of entomostracan and malacostracan neuronal stem cells remains uncertain. It is now well established that during arthropod development, identical and most likely homologous structures can emerge, although the initiating steps or the mode of generation of these structures are different. Recent evidence suggests that adult Entomostraca and Malacostraca share corresponding sets of neurons so that the present report provides an example that those homologous neurons may be generated via divergent developmental pathways. In this perspective, it remains difficult at this point to discuss the question of common patterns of stem cell proliferation with regard to the phylogeny and evolution of Atelocerata and Crustacea. PMID- 11440251 TI - Animal phylogeny and the ancestry of bilaterians: inferences from morphology and 18S rDNA gene sequences. AB - Insight into the origin and early evolution of the animal phyla requires an understanding of how animal groups are related to one another. Thus, we set out to explore animal phylogeny by analyzing with maximum parsimony 138 morphological characters from 40 metazoan groups, and 304 18S rDNA sequences, both separately and together. Both types of data agree that arthropods are not closely related to annelids: the former group with nematodes and other molting animals (Ecdysozoa), and the latter group with molluscs and other taxa with spiral cleavage. Furthermore, neither brachiopods nor chaetognaths group with deuterostomes; brachiopods are allied with the molluscs and annelids (Lophotrochozoa), whereas chaetognaths are allied with the ecdysozoans. The major discordance between the two types of data concerns the rooting of the bilaterians, and the bilaterian sister-taxon. Morphology suggests that the root is between deuterostomes and protostomes, with ctenophores the bilaterian sister-group, whereas 18S rDNA suggests that the root is within the Lophotrochozoa with acoel flatworms and gnathostomulids as basal bilaterians, and with cnidarians the bilaterian sister group. We suggest that this basal position of acoels and gnathostomulids is artifactal because for 1,000 replicate phylogenetic analyses with one random sequence as outgroup, the majority root with an acoel flatworm or gnathostomulid as the basal ingroup lineage. When these problematic taxa are eliminated from the matrix, the combined analysis suggests that the root lies between the deuterostomes and protostomes, and Ctenophora is the bilaterian sister-group. We suggest that because chaetognaths and lophophorates, taxa traditionally allied with deuterostomes, occupy basal positions within their respective protostomian clades, deuterostomy most likely represents a suite of characters plesiomorphic for bilaterians. PMID- 11440252 TI - Multiple origins of a major novelty: moveable abdominal lobes in male sepsid flies (Diptera: Sepsidae), and the question of developmental constraints. AB - Contrary to the impression given by their extreme scarcity among extant species of flies, moveable processes on the abdomen are apparently of relatively simple developmental origin, and they have evolved multiple times in males of the small family Sepsidae. They are used to stimulate the female during copulation in two groups, where they are probably independently derived. Because female cuing of reproductive decisions on particular types of stimuli will tend to favor male abilities to elaborate such stimuli, sexual selection by female choice may sometimes result in sustained selection for certain types of innovations in males. The lack of moveable appendages in most dipterans may be due not to developmental constraints, but to lack of selective advantages. PMID- 11440253 TI - Electronic submission of academic works: a survey of current editorial practices of radiologic journals. AB - Computers are nearly ubiquitous in academic medicine, and authors create and compile much of their work in the electronic environment, yet the process of manuscript submission often fails to utilize the advantages of electronic communication. The purpose of this report is to review the submission policies of major academic journals in the field of radiology and assess current editorial practices relating to electronic submission of academic works. The authors surveyed 16 radiologic journals that are indexed in the Index Medicus and available in our medical center library. They compared the manuscript submission policies of these journals as outlined in recent issues of the journals and the corresponding worldwide web sites. The authors compared the journals on the following criteria: web site access to instructions; electronic submission of text, both with regard to initial submission and final submission of the approved document; text hardcopy requirements; word processing software restrictions; electronic submission of figures, figure hardcopy requirements; figure file format restrictions; and electronic submission media. Although the trend seems to be toward electronic submission, there currently is no clear-cut standard of practice. Because all of the journals that accept electronic documents also require a hardcopy, many of the advantages gained through electronic submission are nullified. In addition, many publishers only utilize electronic documents after a manuscript has been accepted, thus utilizing the benefits of digital information in the printing process but not in the actual submission and peer review process. PMID- 11440254 TI - Opinion: is it time for 'PACSter'? PMID- 11440255 TI - Beam hardening correction for computed tomography images using a postreconstruction method and equivalent tissue concept. AB - A postreconstruction method for correcting the beam-hardening artifacts in computed tomography (CT) images is proposed. This method does not require x-ray spectrum measurement. The authors assumed that a pixel in a CT image can be decomposed into equivalent tissue percentages, depending on its CT number. A scout view of the step wedges made of these equivalent tissues was performed to obtain a beam-hardening correction curve for each tissue. Projecting through the CT image from various angles generated simulated projection data and the total thickness of each tissue along the ray. The correction term was estimated using the tissue thickness traveled by the ray, and this term was then added to its corresponding projection data. A second reconstruction using the corrected projection data yielded a beam-hardening corrected image. The preliminary results show that this method reduces beam hardening artifacts by 14% for aluminum and increased the object contrast by 18% near the aluminum-water boundary. The variation in CT numbers at different locations were reduced, and the aluminum CT number also was restored. PMID- 11440256 TI - Filmless in 60 days: the impact of picture archiving and communications systems within a large urban hospital. AB - Many large urban hospitals converting to filmless radiography use a phased approach for digital imaging implementation. In fact, this strategy often is recommended by picture archival communication systems (PACS) experts and vendors alike for large, busy hospitals installing PACS in existing physical facilities. The concern is that comprehensive conversion from film-based to digital imaging may be too overwhelming an adjustment in operations for a medical staff to effectively handle without serious disruption of workflow for patient treatment and care. Elmhurst Hospital Center is a 543-bed hospital located in the Borough of Queens in New York City. Owned by the New York City Health and Hospitals Corporation, this municipal teaching hospital provides services to a patient mix that is 38% indigent with no insurance, 50% covered by Medicaid or Medicare, and 12% affiliated with HMOs. Most inpatients are admitted through the emergency department. Forty-five percent of all radiology procedures conducted are for emergency patients. Historically, up to 25% of all diagnostic imaging examinations were never reported formally by radiologists. Report turnaround time for the remaining 75% was unacceptable, with only 3% of all imaging examinations reported within a 12-hour period in 1996. Both situations existed in great part because physicians and residents who felt they needed access to films simply took them. Many were never located or returned days after they were taken. In 1998, Elmhurst Hospital Center replaced its RIS and added voice recognition dictation capabilities in January 1999. A hospitalwide PACS was deployed 10 months later. With the exception of mammography, the hospital converted to filmless radiography within 60 days. The critical objectives to maintain control of films and radically improve the reporting process were achieved immediately. Over 99% of all examinations now are formally reviewed and reported. Only 7% of all reports take 1 or more days to generate. This report describes Elmhurst Hospital's efforts to make improvements in the delivery of radiology services and the reasons attributed to its rapid conversion to becoming a filmless (mammography excluded) medical center. The impact of the PACS on radiology department operations and service is discussed. PMID- 11440257 TI - Requirements for an enterprise digital image archive. AB - This report describes several image archival problems facing the authors' department and the results of their attempt to define the requirements for an enterprise digital image archive. The problems identified include the costs of supporting multiple distinct archives, the increased complexity of supporting multiple archive interfaces, the differences in data handling policies and resulting variations in data integrity, and variability in support for nonimage data. The authors also describe the data collected including image volumes and trends and imaging device trends. Finally, the resulting specification for an enterprise digital image archive, including storage and retrieval performance and interface requirements are presented. PMID- 11440258 TI - Determining the sharpness of electronic image displays: an evaluation of three methods. AB - The authors evaluated 3 methods developed to assess the level of monitor cathode ray tube (CRT) sharpness. Results include a comparison of 2 observer-based methods to a charged coupled device (CCD) digital camera-based method for the purposes of CRT equipment comparison, acceptance testing, and routine CRT quality control. Three methods designed to measure a monitor's sharpness were evaluated on a single 20-inch CRT monitor. We defined signal-to-noise ratio (SNR) to be the overall signal difference measured by each method from the highest to lowest values divided by the average standard deviation of the measurements. Comparing the results of the 3 methods, the authors found that the digital CCD camera-based method provided a much higher SNR than the 2 observer-based methods and, therefore, is the preferred of the 3 methods for measuring the sharpness of CRT displays. The SNR values for the CCD, Cx and line target methods were 151.5, 4.9, and 4.3, respectively. The Cx target observer-based method (a novel target and scoring routine dubbed the "Cx" target because of its appearance) had a higher SNR than the line target observer-based method. The average time and standard deviation required to score the Cx and the line targets were 5.45 +/- 2.15 minutes and 8.34 +/- 2.95 minutes, respectively. The observer-based method results (and variability) versus the camera-based method results (and variability) indicate strong linear relationships. Exploring this finding and the optimization of the camera-based method are the subjects of future research. PMID- 11440259 TI - Factors affecting cathode ray tube display performance. AB - Monitors are a complex combination of analog circuits that perform absolutely amazing feats of current and voltage control at both low and high frequency. Digital circuits can reduce the complexity of interfacing with the adjustments and even be used to build complex waveforms, but they cannot replace the point at which pure power is required. Five main topic areas are covered, each building on the other in how they relate to display performance. They are glass composition, resolution and addressability, phosphor selection and aging characteristics, light output and uniformity, and the tools available to assess display performance. PMID- 11440260 TI - Postoperative management of transtibial amputations in VA hospitals. AB - Rigid plaster dressings and immediate postoperative prostheses (IPOP) in patients undergoing transtibial amputations have been reported to reduce pain and healing time, prevent knee flexion contractures, and expedite early ambulation compared to soft dressings. Yet, despite the reported benefits, surgical adoption of (conventional) rigid dressings and IPOP has been inconsistent. The purpose of this study was to determine the current postoperative transtibial amputation dressing practices in VA hospitals. A six-item questionnaire was sent to 134 surgeons at the 117 VA hospitals where transtibial amputations were performed in fiscal year 1999. Responses were received from 83% of the surgeons. During the 1999 study year, surgeons performing transtibial amputations used soft dressings on 67% of patients, conventional rigid dressings with no intent to apply a foot attachment on 14% of patients, removable rigid dressings on 14% of patients, and IPOP (almost exclusively without a foot) on 5% of patients. The application of a rigid dressing or IPOP did not correlate well with the total number of transtibial amputations performed by the surgeon, hospital bed size, or academic affiliation. PMID- 11440261 TI - Mechanical properties of prosthetic limbs: adapting to the patient. AB - Lower-limb amputees have identified comfort and mobility as the two most important characteristics of a prosthesis. While these in turn depend on a multitude of factors, they are strongly influenced by the biomechanical performance of the prosthesis and the loading it imparts to the residual limb. Recent years have seen improvements in several prosthetic components that are designed to improve patient comfort and mobility. In this paper, we discuss two of these: VSAP and prosthetic foot-ankle systems; specifically, their mechanical properties and impact on amputee gait are presented. PMID- 11440262 TI - A comparison of diabetic foot ulcer patients managed in VHA and non-VHA settings. AB - OBJECTIVE: To compare patients with diabetes and new onset foot ulcers treated in Veterans Health Administration (VHA) and non-VHA settings. METHODS: The treatment of patients with new onset diabetic foot ulcers was prospectively monitored in three VHA and three non-VHA hospitals and outpatient settings until ulcer healing, amputation, or death. RESULTS: Of the 302 individuals enrolled in this study, 47% were veterans receiving VHA care. There were no significant differences between veterans and nonveterans in baseline wound classification, diabetes severity, or comorbid conditions. Veterans received significantly fewer sharp debridements, total contact casts, and custom inserts than their nonveteran counterparts, and they had significantly more x-rays, local saline irrigations, IV antibiotics, and prescriptions for bed rest. The percentage of amputations was higher in veterans but did not achieve statistical significance. CONCLUSIONS: Many commonly held stereotypes of veteran men were not found. Veterans and nonveterans with foot ulcers were similar in terms of health and foot history, diabetes severity, and comorbid conditions. There was considerable variation in treatment of diabetic foot ulcers between VHA and non-VHA care. Yet this variation did not result in statistically significant differences in ulcer outcomes. PMID- 11440263 TI - Recreational activities of lower-limb amputees with prostheses. AB - Ninety-two (92) persons with lower-limb amputations who regularly used prostheses responded to a survey that included questions about preferred recreational activities. This article describes the variety of activities selected by these men and women aged 20 to 87 years. Of the activities that were of high importance, 74% to 88% could be performed. Those activities assigned moderate to low importance were less often reported as able to be performed. The activities that require high energy level were more problematic for performance. The diversity of identified activities (n= 166) underscores the value of learning about amputees' activity preferences when making prosthetic prescription decisions. PMID- 11440264 TI - The use of CAD/CAM technology in prosthetics and orthotics--current clinical models and a view to the future. AB - This report examines the current clinical uses of CAD/CAM in prosthetics and orthotics. We conducted interviews to contrast patterns of CAD/CAM use in different private practice settings, at two different teaching institutions, and within two large Prosthetic and Orthotic delivery systems. Investigation into these patterns of use has revealed several lessons. First, there currently exist several very different models of use in clinical practice and these different models will most likely continue. The clinical models range from all traditional techniques with no use of CAD/CAM, to full in-house suites of CAD/CAM equipment with extensive utilization, to a simplified office with minimal in-house equipment and minimal fabrication and a near total dependency on central fabrication. Second, a growing number of prosthetic and orthotic devices are successfully being fabricated and fit with CAD/CAM technology after starting the process with simple measurements instead of casted, scanned, or digitized exact anatomic data. Starting the CAD process with "by-the-numbers approach" has revealed the reality that for some devices, the "input" needed to define the shape of the residual limb or torso, may not need to be as accurate as originally thought. Third, the fabrication techniques that are currently being used with CAD/CAM systems are still rather traditional techniques. Most devices are still laminated or formed over computer carved models. Although research continues into advanced fabrication techniques, the prosthetics and orthotics industry has not yet taken advantage of the possibilities in the computer-assisted manufacturing side of the equation. Finally, the business of manufacturing and selling up-to date CAD/CAM equipment and software has a tremendous impact on how this technology is used in prosthetics and orthotics. The size of the prosthetics and orthotics industry and the potential number of customers for major equipment are relatively small. Being in the business of providing the advanced CAD/CAM systems of tomorrow is an expensive and difficult proposition. Current users of full in house CAD systems have expressed concern that upgrading equipment and software might not be economical with today's decreasing revenues. For all of these reasons, many believe that the number of practitioners who use a central fabrication model will grow more rapidly than the number of practitioners who own and operate a full in-house system. PMID- 11440265 TI - Can the presence of equinus contracture be established by physical exam alone? AB - The condition in which ankle dorsiflexion is restricted is known as equinus contracture (EC). Equinus contracture is purported to be associated with a number of clinical conditions. However, there are no data to support or refute a clinician's ability to diagnose EC by clinical exam. We prospectively evaluated the maximum ankle dorsiflexion with the knee fully extended in 68 people (34 patients with isolated fore- or midfoot pain and 34 asymptomatic subjects) both by clinical exam and by a custom-designed ankle goniometer. We compared the likelihood of agreement of the clinical impression (equinus, no equinus) to the maximum ankle dorsiflexion measured with the instrument at two different numerical definitions of EC (< or =5 degrees and < or =10 degrees of maximum dorsiflexion). When all subjects were included and equinus defined as < or =5 degrees of ankle dorsiflexion, a clinician's ability to detect the equinus when it is truly present is 77.8%. If equinus is defined as < or =10 degrees, this ability increases to 97.2%. Alternatively, if equinus is not present, as defined by < or =5 degrees, then a clinician's ability to correctly diagnose no equinus is 93.8%. If equinus is defined to < or =10 degrees, this ability decreases to 68.8%. PMID- 11440266 TI - Survival following lower-limb amputation in a veteran population. AB - GOAL: We sought to describe the common demographic and comorbid conditions that affect survival following nontraumatic amputation. METHODS: Veterans Administration hospital discharge records for 1992 were linked with death records. The most proximal level during the first hospitalization in 1992 was used for analysis. Demographic information (age, race) and comorbid diagnosis (cardiovascular, cerebrovascular, and renal disease) were used for Kaplan-Meier curves to describe survival following amputation. MAIN OUTCOME MEASURE: Death. RESULTS: Mortality risk increased with advanced age, more proximal amputation level, and renal and cardiovascular disease, and decreased for African Americans. No increased risk for persons with diabetes was noted in the first year following amputation but the risk increased thereafter. A higher risk of mortality in the first year was noted for renal disease, cardiovascular disease, and proximal amputation level. CONCLUSION: Survival following lower-limb amputation is impaired by advancing age, cardiovascular and renal disease, and proximal amputation level. Also, a small survival advantage is seen for African Americans and those with diabetes. PMID- 11440267 TI - Trends in peripheral vascular procedures in the Veterans Health Administration, 1989-1998. AB - OBJECTIVE: To assess trends in peripheral vascular procedures performed in Veterans Health Administration (VHA) facilities. METHODS: All discharges with peripheral vascular procedures recorded for 1989-1998 were analyzed. The VHA user population was used to calculate age-specific rates. Trends were evaluated using frequency tables and Poisson regression. RESULTS: The VHA had 55,916 discharges with peripheral vascular procedures performed almost exclusively in men. Indications included peripheral vascular disease (53.7%), gangrene (19.3%), surgical complications (13.3%), and ulcers and infection (9.6%). The VHA age specific rates were higher than US population rates for persons 45 to 64 years, similar for those 65 to 74 years, and lower for those 75 years and older. The age specific rates declined slightly over the 10 years of observation, with the greatest decline noted in men age 45 to 65. CONCLUSION: The VHA provides almost 8% of all US peripheral vascular procedures in males. The VHA age-specific rates differ from the US rates with a shift to younger patients. The rates decreased for all age groups between 1989-1998. PMID- 11440268 TI - Limb loss prevention and prosthetic engineering. PMID- 11440269 TI - Tribute to Ernest M. Burgess, MD, PhD October 29, 1911-September 27, 2000. PMID- 11440270 TI - Regulation of renal growth and IGFBP-4 expression by triiodothyronine during rat development. AB - The insulin-like growth factors (IGFs) and insulin-like growth factor binding proteins (IGFBPs), which regulate IGF activity, play a fundamental role in renal cell proliferation and differentiation. The thyroid hormone is considered to be required for kidney development; excess induces local hypertrophy and hyperplasia. The aim of the present study was to investigate the possible involvement of the IGF/IGFBP system in thyroid hormone-induced renal growth during the development of the rat. Our results show that thyroid hormone withdrawal by 6-propyl-2-thiouracil (PTU)-treatment of rats at all ages had no effect on renal IGFBP-4 mRNA levels, whereas the abundance of the serum protein was decreased compared to controls. Intraperitoneal triiodothyronine (T3) administration to hypothyroid rats resulted in renal hypertrophy associated with a significant upregulation of IGFBP-4 expression with increased levels of renal IGFBP-4 mRNA and serum protein. T3-induced upregulation of IGFBP-4 expression suggests the involvement of the local IGF/IGFBP system in T3-induced renal hypertrophy. PMID- 11440271 TI - Mutations in the mouse TSH receptor equivalent to human constitutively activating TSH receptor mutations also cause constitutive activity. AB - Constitutively activating mutations in the human thyroid-stimulating hormone (TSH) receptor (TSHr) have been identified as the most prevalent molecular cause of non-autoimmune hyperthyroidism. To investigate the feasibility of an animal model for non-autoimmune hyperthyroidism, we introduced two mutations in the mouse TSHr which had previously been identified in the human TSHr. The two human mutations showed strong differences in TSH binding, basal cAMP and IP accumulation. In the human TSHr, the Ile 486 Phe mutation causes a high increase of basal cAMP accumulation and also basal stimulation of the inositol phosphate pathway, whereas the Val 509 Ala mutation results in a low increase of basal cAMP accumulation without affecting IP signaling. RNA was isolated from mouse thyroid tissue and reverse transcribed. A 2.4 kb PCR product from the mouse TSHr was cloned into the pGEM-T vector system. Ile was substituted with Phe at codon 486 and Val with Ala at codon 509. These mutated mouse TSHrs were subcloned in the pSVL expression vector. After transient expression in COS-7 cells, basal and TSH stimulated cAMP and IP accumulation, cell surface expression and TSH binding were determined and directly compared to the human TSHr. Whereas constitutively activating mutations of the human parathyroid hormone (PTH)/PTH-related peptide receptor showed little or no change in basal cAMP accumulation when introduced into the rat PTH/PTHrP receptor, these two mouse TSHr mutations resulted in constitutive activity similar to the homologous mutations in the human TSHr. Therefore, it should be possible to establish a mouse model for non-autoimmune hyperthyroidism by homologous recombination to study the pathogenetic mechanisms of non-autoimmune hyperthyroidism. PMID- 11440272 TI - Involvement of L-triiodothyronine in acetylcholine metabolism in adult rat cerebrocortical synaptosomes. AB - Despite the recently emerging notion of thyroid-hormone involvement in neurotransmission in the adult mammalian brain, adequate evidence for a cellular basis of the process is still lacking. The present study indicates the involvement of thyroid hormones in cholinergic system of the adult rat cerebral cortex. Administration of L-triiodothyronine (T3, 0.025 to 4 microg/g) in single doses increased the synaptosomal acetylcholinesterase (AchE) and Mg2+-ATPase activity maximally at 24 hours in a dose-dependent way. Propylthiouracil (PTU) treated hypothyroid rats showed a significant increase in AchE and Mg2+-ATPase activity compared to euthyroid rats. T3-treatment on hypothyroid rats decreased AchE activity in synaptosomes compared to the hypothyroid synaptosomal values. Mg2+-ATPase activity found in (PTU + T3)-treated group and T3-treated group remained high. These results predict that T3 stimulates acetylcholine (Ach) metabolism by increasing AchE activity as well as uptake of the released Ach through an increase in synaptosomal Mg2+-ATPase activity. This indicates a positive impact of T3 on the cholinergic system in the adult mammalian brain. PMID- 11440273 TI - The effects of fasting and refeeding on serum parathormone and calcitonin concentrations in young and old male rats. AB - Although fasting and refeeding reveal the existence of age-related changes in carbohydrate and lipid metabolism, the effects of aging on mineral metabolism in refed animals are unknown. We therefore investigated hormonal regulation of calcium metabolism in young (4 months) and old (26 months) male rats fasted for 48 hours and then refed for 4 or 24 hours. Serum concentrations of total and ionized calcium and parathormone were similar in control young and old rats. Serum calcitonin level was higher, and the concentrations of albumin and inorganic phosphate and alkaline phosphatase activity were lower in fed old rats. In young fasted rats, the serum ionized and total calcium was decreased, and phosphate concentration was increased. In old rats, fasting resulted in the increase of serum parathormone level. Fasting reduced serum alkaline phosphatase activity to a similar extent in both age groups. In young rats, refeeding for 24h normalized serum calcium and phosphate levels and alkaline phosphatase activity, and decreased serum concentrations of PTH and calcitonin. In old refed rats, serum calcitonin concentration was raised by 77% compared to fed or fasted animals, whereas parathormone levels were normalized. Our results indicate that old fasted or refed rats maintain normal serum calcium concentration in a different way than young animals, possibly through the increase in serum levels of parathormone and/or calcitonin. Thus, dietary manipulations such as fasting and refeeding constitute an interesting model for the investigation of the effects of aging on the hormonal regulation of serum calcium level. PMID- 11440274 TI - Effect of electrical field stimulation on insulin and glucagon secretion from the pancreas of normal and diabetic rats. AB - The effect of electrical field stimulation (EFS) on insulin (INS) and glucagon (GLU) secretion from normal and diabetic rat pancreas is poorly understood. In our study, EFS (5-20Hz, 50 V amplitude and 1.0 ms pulse width), when applied alone, resulted in a significant (p<0.05) increase in INS secretion from the pancreas of both normal and diabetic rats. Atropine (10(-5) M) did not inhibit the EFS (5 Hz)-evoked INS secretion in normal pancreas and failed to alter the effect of EFS (10-20 Hz) on INS secretion from the pancreas of both normal and diabetic rats. Propranolol (Prop) inhibited INS secretion to below basal level in the presence of EFS (5 Hz) but not at EFS (10- 20 Hz). Tetrodotoxin (TTX) also significantly (p = 0.002) inhibited INS secretion from normal pancreas in the presence of EFS (5-20 Hz). The decrease in insulin secretion observed when pancreatic tissue fragments were incubated in Prop and TTX in the presence of EFS was reversed by yohimbine (10(-5) M). In contrast, TTX did not significantly modify INS secretion from diabetic pancreas in the presence of EFS. EFS (5-20 Hz) significantly (p<0.05) increased GLU release from normal and diabetic rat pancreas when applied alone. Neither atropine, Prop nor TTX significantly modified GLU release from the pancreas of either normal or diabetic rats. This suggests that GLU secretion may be controlled through a different pathway. The EFS-evoked INS and GLU secretion is probably executed via different mechanisms. These mechanisms include 1) activation of cholinergic nerves by EFS; 2) EFS of alpha- and beta-adrenergic nerves; 3) activation of non-adrenergic non cholinergic pathway by EFS; 4) EFS-induced depolarization and subsequent action potential in pancreatic endocrine cells and 5) electroporosity caused by EFS induced membrane permeability. All of these effects may be summative. In conclusion, EFS (5-20 Hz), when applied alone, can evoke significant increases in INS and GLU secretion from the pancreas of both normal and diabetic rats. Insulin secretion is controlled via alpha-2 adrenergic (inhibition) and beta-adrenergic (stimulation) receptors. Glucagon secretion is enhanced by alpha2 adrenergic stimulation. PMID- 11440275 TI - Plasma adrenomedullin concentrations in patients with adrenal pheochromocytoma. AB - BACKGROUND: The hypotensive peptide adrenomedullin was first isolated in extracts of human pheochromocytoma. There is, however, no information available on the behaviour of circulating adrenomedullin or on the correlation with catecholamines in patients with pheochromocytoma. OBJECTIVES: 1) to investigate whether plasma adrenomedullin levels were changed in 10 patients with pheochromocytoma when compared to 21 healthy subjects and 16 patients with essential hypertension; 2) to determine whether or not adrenomedullin has a counter-regulatory role in catecholamine excess in pheochromocytoma or is responsible for hemodynamic modifications before and after tumour resection; 3) to determine tissue distribution of iradrenomedullin in the pheochromocytoma. METHODS: Plasma adrenomedullin and catecholamine levels were measured in all patients with pheochromocytoma before and four weeks after tumour removal. In the four patients undergoing resection of tumours, plasma levels of adrenomedullin were measured at different time-points during surgery. RESULTS: The mean plasma adrenomedullin concentrations ( SD) in patients with pheochromocytoma (37.9 +/- 6pg/ml) were significantly higher (p<0.0001) than those in normal subjects (13.7 +/- 6.1 pg/mI) and patients with essential hypertension (22.5 +/- 9.lpg/ml). Adrenomedullin levels correlated with plasma noradrenaline (r = 0.516, p = 0.0124). In all patients with pheochromocytoma, plasma adrenomedullin concentrations decreased after removal of tumours (from 37.9 +/- 6 to 10.9 +/- 4.6 pg/ml; p < 0.0001). In the four patients studied during surgery, baseline plasma adrenomedullin and noradrenaline levels were markedly elevated, and increased significantly with tumour manipulation, decreasing 24 hours after operation. Adrenal medulla cells surrounding the pheochromocytoma site stained for ir-adrenomedullin, whereas only isolated cells of pheochromocytoma stained for the peptide. CONCLUSIONS: This study demonstrates that circulating adrenomedullin is increased in pheochromocytoma, and is also correlated with plasma noradrenaline levels. Adrenomedullin may represent an additional biochemical parameter for clinical monitoring of patients with pheochromocytoma. PMID- 11440276 TI - The therapeutic problem of proliferative diabetic retinopathy: targeting somatostatin receptors. AB - Clinical management of proliferative diabetic retinopathy has changed very little in the last 5 decades, relying primarily on laser ablation of the retinal vasculature. Several lines of clinical and experimental evidence suggest that somatostatin analogues may be efficacious in inhibiting neovascularization associated with proliferative retinopathy but the mechanism of action for these compounds is unclear. Inhibition of growth hormone secretion and the subsequent suppression of insulin-like growth factor 1 (IGF-1) production by somatostatin has been suggested as the mechanism of action, however, in vitro studies suggest that somatostatin analogues suppress endothelial cell growth through a direct, somatostatin receptor-mediated inhibition of pro-survival signaling pathways. The advent of a new generation of modified peptide and peptidomimetic somatostatin analogues has allowed investigators to more carefully define the receptor subtypes responsible for somatostatin-induced endothelial cell death and may eventually lead to the clinical development of somatostatin analogues that can reduce endothelial cell proliferation, independent of suppression of circulating hormone levels. PMID- 11440277 TI - Octreotide reduces vitreous hemorrhage and loss of visual acuity risk in patients with high-risk proliferative diabetic retinopathy. AB - OBJECTIVE: Growth hormone and insulin-like growth factor-I have been implicated as strong promoters of proliferative diabetic retinopathy. We studied reduction of bleeding and preservation of visual acuity by treatment with the long-acting somatostatin analogue, octreotide, in diabetic patients at an advanced stage of proliferative diabetic retinopathy. RESEARCH DESIGN AND METHODS: Randomized trial in a University hospital setting. Reading ophthalmologists were masked for octreotide use, diabetologists were aware of that treatment. Nine patients received 100 microg tid octreotide (verum) subcutaneously for a maximum of 36 months. Nine diabetics served as controls, no placebo treatment was used. Episodes of vitreous hemorrhages were counted, measurement of visual acuity, estimation of neovascularization by stereoscopic fundus photography and fluorescein angiography were carried out. RESULTS: After 3 years of treatment, the incidence of vitreous hemorrhages and the need for vitreoretinal surgery was significantly lower (log rank test p = 0.002) in the octreotide-treated patients. Visual acuity was preserved and significantly better in the octreotide treated group compared to controls (p = 0.05). CONCLUSIONS: In diabetics with high-risk proliferative retinopathy after full scatter laser coagulation, octreotide reduced the number of vitreous hemorrhages, preserving visual acuity. PMID- 11440278 TI - Topical application of integrin antagonists inhibits proliferative retinopathy. AB - The expression of alphav-integrins is highly selective for angiogenic endothelial cells; ligation inhibition by cyclic RGD peptides prevents pathological neovascularization in tumor or retinopathy models to a large extent. We have previously demonstrated that proliferative retinopathy in a mouse model of retinopathy of prematurity (ROP model) can be reduced by more than 70%. To minimize systemic side effects and unwanted interference with responsive angiogenesis, we investigated topical application of cyclic RGD-peptides. In preliminary experiments, we could exclude any inhibiting effects of the carrier solution containing EDTA, Na2S, mannitol, hydroxyethyl starch, and benzalconium chloride on the inhibitory effect of cyclic RGD peptides. Retinal presence of small molecular-mass integrin antagonists after topical application was confirmed using fluorescein-labeled cyclic RGD peptide. Topical application of the peptide to the eye inhibited proliferative retinopathy in a dose-dependent fashion with a maximum of almost 50%. These results suggest that small molecular-mass peptide antagonists of alphav-type integrins are efficient in inhibiting proliferative retinopathy by topical application. PMID- 11440279 TI - Retinal photocoagulation does not influence intraocular levels of IGF-I, IGF-II and IGF-BP3 in proliferative diabetic retinopathy-evidence for combined treatment of PDR with somatostatin analogues and retinal photocoagulation? AB - Retinal photocoagulation reduces the incidence of severe visual loss in proliferative diabetic retinopathy (PDR). Reduced levels of VEGF/VPF might result in an improved function of the blood-retina barrier and cause a decrease of blood derived intraocular growth factors such as IGF-I. This study investigates whether retinal photocoagulation is able to normalize the concentrations of IGF-I, IGF-II and IGF-BP3 in the vitreous humor of patients undergoing vitrectomy. Levels of IGFs and the permeability marker, albumin, were measured in serum and vitreous of 52 patients. Three groups were compared: controls without proliferating eye disease (n = 19) and patients with PDR with (PDR+; n = 25) and without (PDR-; n = 8) previous retinal photocoagulation. IGF-I, IGF-II, IGF-BP3 and albumin were determined by immunological methods and were confirmed to be increased in patients with PDR compared to controls. Retinal photocoagulation influenced neither the intraocular concentration of the permeability marker albumin (PDR+: 253.2 +/- 46 mg/dl; PDR-: 256.4 +/- 66.5 mg/dl) nor the levels of IGFs (PDR+: IGF I: 1.2 +/- 0.1 ng/ml; p = 0.38; IGF-II: 34.8 +/- 2.2 ng/ml; p = 0.1; IGF-BP3: 75.7 +/- 9.7 ng/ml; p = 0.27; PDR-: IGF-I: 1.1 +/- 0.2ng/ml; IGF-II: 29.3 +/- 5.2 ng/ml; IGF-BP3: 61.5 +/- 18.3 ng/ml). Systemic levels of albumin and IGFs were not changed significantly by retinal photocoagulation. These results demonstrate that previous retinal photocoagulation in patients undergoing vitrectomy does not functionally reestablish the blood-retina barrier despite decreases in VEGF/VPF. The lack of influence on intraocular concentrations of the serum-derived growth factors, IGF-I, IGF-II and IGF-BP3, might in part explain the failure of previous photocoagulation in the investigated patients. These results suggest that a combined treatment with retinal photocoagulation and growth hormone-lowering drugs, such as somatostatin analogues, could be a useful treatment, which may prevent further loss of visual acuity in patients with PDR. PMID- 11440280 TI - Increased levels of vascular endothelial growth factor and advanced glycation end products in aqueous humor of patients with diabetic retinopathy. AB - Clinical studies have shown a relationship between diabetic retinopathy and vascular endothelial growth factor (VEGF) levels in ocular fluid. Advanced glycation end products (AGEs) have been implicated in diabetes complications, including diabetic retinopathy. Nepsilon-(carboxymethyl) lysine (CML) is a glycoxidation product that may be a marker of oxidative stress. In this study, we used enzyme-linked immunosorbent assays to determine the levels of VEGF, non-CML AGE and CML in the aqueous humor and serum of 82 Japanese patients with type 2 diabetes and 60 non-diabetic subjects. VEGF, non-CML AGE, and CML concentrations in aqueous humor and serum were then compared with the severity of diabetic retinopathy. Immunohistochemical detection analysis of non-CML AGE and CML was also performed using retinal tissues from patients with progressive diabetic retinopathy. Aqueous levels of VEGF, non-CML AGE and CML increased along with the progression of diabetic retinopathy compared to age-matched controls. After coagulation therapy, the VEGF, non-CML AGE, and CML levels were significantly reduced. Immunostaining showed diffuse co-localization of non-CML AGE and CML around microvessels and in the glial cells of proliferative membranes from patients with progressive diabetic retinopathy. These findings suggest that glycation and glycoxidation reactions (or oxidation, as revealed by CML) may contribute to both the onset and progression of diabetic retinopathy. PMID- 11440281 TI - Current options for the prevention and treatment of migraine. AB - BACKGROUND: Migraine is a common condition affecting approximately 18% of women and 6% of men in the United States. The goals of managing migraine are 2-fold: to prevent attacks from occurring and to effectively and rapidly end them when they do occur. OBJECTIVE: This article reviews the acute and prophylactic treatment of migraine. METHODS: Information for inclusion in this review was identified through a search of MEDLINE from 1995 to the present. Search terms included migraine, acute treatment, prophylactic treatment, preventive treatment, and individual drug names. RESULTS: Preventive measures for migraine include lifestyle changes (eg, avoiding migraine triggers and maintaining regular sleep, eating, and work habits) and drug therapy. Beta-blockers, calcium channel blockers, tricyclic antidepressants, and anticonvulsants are among the more common drug classes used for migraine prophylaxis, but preventive therapy must be individualized, taking into account efficacy, potential adverse effects, co existing medical conditions, and drug costs. Many medications are available for the acute treatment of migraine, including over-the-counter analgesics and prescription drugs. Of the latter, the 5-hydroxytryptamine(1B/1D)-receptor agonists, or triptans, are the most recently introduced class. Each of the 4 available triptans (sumatriptan, zolmitriptan, naratriptan, and rizatriptan) is effective in ending a migraine attack, but comparative trials have shown differences between individual drugs in the time to pain relief and the percentage of patients who obtain pain relief. CONCLUSIONS: Medications to prevent or reduce the frequency of migraine tend to be less specific and effective than medications for the acute treatment of migraine. As a class, triptans are generally well tolerated and may be considered drugs of choice for the acute treatment of moderate to severe migraine. PMID- 11440282 TI - Review of oxymetholone: a 17alpha-alkylated anabolic-androgenic steroid. AB - BACKGROUND: Oxymetholone (17beta-hydroxy-2-[hydroxymethylene]-17-methyl-5alpha androstan-3-one) is a 17alpha-alkylated anabolic-androgenic steroid and a synthetic derivative of testosterone. It has been approved by the US Food and Drug Administration for the treatment of anemias caused by deficient red cell production. OBJECTIVES: This review summarizes the pharmacokinetics, current and future clinical applications, and adverse effects of oxymetholone. Relevant studies were identified using a search of MEDLINE through March 2001, supplemented by conference abstracts and presentations. RESULTS: Because of its anabolic properties, oxymetholone has been studied for the treatment of HIV associated wasting, antithrombin III deficiency, pediatric growth impairment, and damaged myocardium, with varying degrees of success. Hepatotoxicity is a major adverse effect associated with the use of oxymetholone, with cholestatic jaundice the most important hepatic side effect. Less common hepatic side effects associated with the use of anabolic-androgenic steroids include peliosis hepatis and formation of hepatic tumors. All anabolic-androgenic steroids can cause androgenic side effects, including acne, hirsutism, hair loss, clitoral/phallic enlargement, vocal changes, erectile tissue stimulation, gynecomastia, amenorrhea, and changes in libido and sexual potency. CONCLUSIONS: As is the case with many anabolic-androgenic steroids, few pharmacokinetic and tolerability studies were performed before oxymetholone's approval in the 1960s. It has proved, however, to be an appropriate treatment choice for selected patients with anemia, if carefully monitored. PMID- 11440283 TI - Entacapone: a catechol-O-methyltransferase inhibitor for the adjunctive treatment of Parkinson's disease. AB - BACKGROUND: When levodopa therapy is used in Parkinson's disease, degradation of the drug in the peripheral nervous system is associated with dyskinesias and motor fluctuations. Much of this degradation is produced by catechol-O methyltransferase (COMT), an enzyme involved in the metabolism of catecholamines and catechol compounds. Inhibition of COMT activity prolongs the action of levodopa and reduces fluctuations in response. Entacapone is a selective inhibitor of COMT whose activity is primarily in the peripheral nervous system, with little effect in the central nervous system. OBJECTIVE: This paper reviews the pharmacologic properties and clinical usefulness of entacapone in the treatment of Parkinson's disease. METHODS: Recent studies, abstracts, and reviews published in the English-language literature were identified through searches of MEDLINE (1966-September 2000), International Pharmaceutical Abstracts (1970 September 2000), and PharmaProjects (September 2000 version), and from the Web sites of Parkinson's disease conferences held from 1996 to September 2000. Relevant human studies provided further information on the pharmacologic properties and clinical usefulness of entacapone. RESULTS: Entacapone is rapidly absorbed, with a time to maximum concentration of approximately 1 hour. Its plasma elimination half-life is 0.4 to 0.7 hour in the beta phase and 2.4 hours in the gamma phase, and it has 35% absolute bioavailability after oral administration, secondary to first-pass clearance. Entacapone is 98% protein bound; thus, it is not distributed widely in tissues and is almost completely metabolized before excretion (0.1%-0.2% of dose unchanged in urine). The drug inhibits erythrocyte-soluble COMT activity in a dose-dependent fashion (48% after a 400-mg dose, 82% after an 800-mg dose). The inhibitory effect is reversible, with recovery of soluble COMT activity within 4 to 8 hours. In levodopa-treated patients with Parkinson's disease who experience motor fluctuations, clinical trials have demonstrated entacapone's effectiveness in increasing "on" time (the period during which medications relieve the symptoms of Parkinson's disease) by up to 1.2 hours, decreasing "off" time (the period during which symptoms increase) by 0.9 to 1.3 hours, and producing overall total improvement in scores on the Unified Parkinson's Disease Rating Scale. The recommended dosage of entacapone is 200 mg administered orally with each dose of levodopa/carbidopa, up to 8 doses per day. The drug is generally well tolerated, with most adverse effects attributed to levodopa-related dopaminergic effects, including dyskinesias and nausea. CONCLUSIONS: In clinical trials, adjuvant treatment with entacapone appeared to be an effective and well-tolerated therapeutic strategy in patients with Parkinson's disease who experience fluctuations in the response to levodopa therapy. The increased elimination half-life of levodopa with concomitant entacapone results in greater and more sustained plasma levodopa levels, with more constant dopaminergic stimulation in the brain and greater amelioration of parkinsonian symptoms. PMID- 11440284 TI - Telmisartan plus hydrochlorothiazide versus telmisartan or hydrochlorothiazide monotherapy in patients with mild to moderate hypertension: a multicenter, randomized, double-blind, placebo-controlled, parallel-group trial. AB - BACKGROUND: Recent surveys reveal continuing deficiencies in the awareness, treatment, and control of hypertension. In many cases, failure to achieve blood pressure targets may be attributable to the use of antihypertensive monotherapy. OBJECTIVES: This study was undertaken to identify combinations of telmisartan, a new oral angiotensin II type 1-receptor antagonist, and hydrochlorothiazide (HCTZ) that might provide greater antihypertensive efficacy than monotherapy with either agent in the treatment of mild to moderate hypertension. It also examined the dose-response surface for the 2 drugs alone and in combination. METHODS: This was a multicenter, randomized, double-blind, placebo-controlled, parallel-group study that employed all cells of a 4 x 5 factorial design. After a 4-week, single blind, placebo run-in period, men and women between 18 and 80 years of age with mild to moderate hypertension (defined as mean supine diastolic blood pressure [DBP] between 95 and 114 mm Hg during the last 2 weeks of the placebo run-in period and systolic blood pressure [SBP] between 114 and 200 mm Hg immediately before randomization) were eligible to enter the 8-week, double-blind, double dummy treatment period. Study comparisons were between once-daily telmisartan monotherapy (20, 40, 80, or 160 mg), HCTZ monotherapy (6.25, 12.5, or 25 mg), 12 combinations of these telmisartan/HCTZ doses, and placebo. The focus was on 2 combinations: telmisartan 40 mg/HCTZ 12.5 mg and telmisartan 80 mg/HCTZ 12.5 mg. The primary efficacy variable was change in supine trough DBP from baseline to the last evaluable measurement during double-blind treatment. Plasma renin activity and safety parameters, including treatment-emergent adverse events, physical findings, electrocardiograms, and serum electrolyte levels (which are known to increase with HCTZ treatment), were also assessed. RESULTS: Of 1293 patients screened, 818 (63.3%) were enrolled at 47 centers. Of these 818, 749 (91.6%) completed the study. The intent-to-treat population (randomized with > or = 1 postrandomization blood pressure measurement) consisted of 807 patients (98.7%). Telmisartan 80 mg/HCTZ 12.5 mg significantly decreased mean supine trough SBP/DBP by 23.9/14.9 mm Hg, a benefit of 8.5/3.4 mm Hg compared with telmisartan 80 mg and of 17.0/7.6 mm Hg compared with HCTZ 12.5 mg (both comparisons, P < 0.01). Telmisartan 40 mg/HCTZ 12.5 mg significantly reduced mean supine SBP by 18.8 mm Hg, a benefit of 6.6 mm Hg compared with telmisartan 40 mg and 11.9 mm Hg compared with HCTZ 12.5 mg (both, P < 0.01). This same combination significantly reduced mean supine DBP by 12.6 mm Hg, a benefit of 5.3 mm Hg compared with HCTZ 12.5 mg (P < 0.01), but was not significantly different from telmisartan 40 mg. Telmisartan 80 mg/HCTZ 12.5 mg was significantly more effective than telmisartan 40 mg/HCTZ 12.5 mg in reducing mean supine DBP and SBP (both, P < 0.05). The response surface and responder analyses confirmed the additive antihypertensive efficacy of the combination of telmisartan and HCTZ. All regimens were well tolerated. CONCLUSIONS: Once-daily telmisartan 80 mg/HCTZ 12.5 mg was effective and well tolerated when used to reduce SBP and DBP in patients with mild to moderate hypertension. In addition to enhancing efficacy, this combination protected against potassium depletion, a common side effect of thiazide monotherapy. PMID- 11440285 TI - Effects of low doses of simvastatin and atorvastatin on high-density lipoprotein cholesterol levels in patients with hypercholesterolemia. AB - BACKGROUND: Simvastatin 40 to 80 mg/d has been found to increase high-density lipoprotein cholesterol (HDL-C) levels significantly more than atorvastatin at equipotent doses (ie, 20-80 mg/d). Data on the effects of lower doses of the 2 drugs on HDL-C levels are conflicting. OBJECTIVE: The purpose of this study was to investigate the effects of simvastatin 20 mg/d and atorvastatin 10 mg/d on HDL C levels in patients with hypercholesterolemia. METHODS: Patients with primary hypercholesterolemia (total cholesterol [TC] >250 mg/dL) who were not taking any lipid-lowering agents and who were following a low-fat diet were randomized to receive 1 of 2 treatments: simvastatin 20 mg/d or atorvastatin 10 mg/d. Serum TC, triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and HDL-C levels were measured using standard methods after 2 months of therapy. In a secondary analysis, lipids and lipoprotein cholesterol were measured after 1 year in patients who continued treatment. RESULTS: Of the 240 patients enrolled (108 men and 132 women; age range, 23-77 years, mean [SEM] 56.7 [0.69]), 235 completed the study. After 2 months of therapy, TC, LDL-C, and serum TG levels decreased significantly versus baseline in both groups (P < 0.001), with no significant differences between treatment groups. HDL-C levels increased by 9.0% (P < 0.001 vs baseline) in the simvastatin group and by 4.3% (P < 0.02) in the atorvastatin group. The difference between the 2 groups in the percentage increase in HDL-C was statistically significant (P < 0.05). In 113 patients who continued treatment, HDL-C levels at 1 year were still significantly higher than baseline levels in the simvastatin group (6.3%, P = 0.034), but not in the atorvastatin group (2.8%, P = 0.587). CONCLUSIONS: The findings from this study suggest that the HDL-C-increasing effect of simvastatin 20 mg is significantly greater than that of atorvastatin 10 mg. Since increasing HDL-C levels is thought to lower the risk for atherosclerosis and coronary heart disease, these results warrant further investigation. PMID- 11440286 TI - STATT: a titrate-to-goal study of simvastatin in Asian patients with coronary heart disease. Simvastatin Treats Asians to Target. AB - BACKGROUND: Most published studies on the use of lipid-lowering agents to treat hypercholesterolemia have focused on Western populations, with few data on Asian populations. OBJECTIVE: The Simvastatin Treats Asians to Target (STATT) study used a titrate-to-goal protocol to evaluate the efficacy and tolerability of simvastatin 20 to 80 mg/d in the treatment of Asian patients with coronary heart disease. METHODS: This was a multicenter, open-label, uncontrolled, 14-week study in patients with coronary heart disease and serum low-density lipoprotein cholesterol (LDL-C) levels of 115-180 mg/dL and triglyceride levels of < or = 400 mg/dL. The dose of simvastatin was titrated from 20 to 80 mg/d to achieve the National Cholesterol Education Program (NCEP) LDL-C target of < or = 100 mg/dL. The primary efficacy measure was the percentage of patients achieving the NCEP target. Among secondary measures were the percentage of patients achieving European Society of Cardiology/European Atherosclerosis Society/European Society of Hypertension target LDL-C levels of < or = 115 mg/dL and the percentage change from baseline in lipid parameters. Tolerability was assessed in terms of the overall incidence of adverse experiences and the incidences of the most commonly reported adverse experiences. RESULTS: The intent-to-treat analysis included 133 Asian patients (93 men, 40 women; mean age, 59.5 years), of whom 125 completed 14 weeks of therapy. Their mean blood pressure was 130.2/79.4 mm Hg. Overall, 104 (78.2%) patients treated with simvastatin achieved LDL-C levels < or = 100 mg/dL at week 14, and 125 (94.0%) achieved this target at some point during the study. Similarly, 122 (91.7%) patients achieved an LDL-C level < or = 115 mg/dL at week 14, and 130 (97.7%) achieved this target at some point during the study. Treatment with simvastatin had favorable effects on the lipid profile, producing significant percentage changes from baseline in all parameters (P < 0.001). Simvastatin was well tolerated across the dose range. Overall, 40 patients (30.1%) had > or = 1 clinical adverse experience. Only 14 (10.5%) had adverse experiences that were possibly, probably, or definitely related to study drug; none of these experiences were considered serious. The most common adverse experiences (> or = 3% incidence) were abdominal pain (6%); chest pain (5%); dizziness (4%); and asthenia/fatigue, fibromyalgia, headache, insomnia, and upper respiratory tract infection (3% each). No new or unexpected adverse experiences were seen at the higher doses. CONCLUSIONS: Simvastatin was effective and well tolerated at doses of 20, 40, and 80 mg/d in Asian patients with coronary heart disease. Titration enabled the majority to achieve target LDL-C levels of < or = 100 mg/dL. PMID- 11440287 TI - A population pharmacokinetic model that describes multiple peaks due to enterohepatic recirculation of ezetimibe. AB - BACKGROUND: Ezetimibe, a selective inhibitor of intestinal cholesterol absorption, is in clinical development for the treatment of hypercholesterolemia. It is rapidly absorbed and glucuronidated in the intestine. The parent compound and its conjugated metabolite undergo enterohepatic recirculation, resulting in multiple peaks in the plasma concentration-time profile. OBJECTIVE: The purpose of this study was to develop a population pharmacokinetic (PPK) model for ezetimibe that incorporates enterohepatic recirculation. METHODS: A population compartment model incorporating input from the gallbladder, consistent with food intake, was developed to account for enterohepatic recirculation. The amount recycled was allowed to vary within a subject and between subjects, accommodating variability in bile secretion. The data used consisted of 90 profiles from healthy subjects who received single or multiple doses of ezetimibe 10 or 20 mg. Modeling was carried out using a nonlinear mixed-effect function in the S-PLUS statistical program. RESULTS: The amount of ezetimibe recycled into the central compartment was estimated to be approximately 17% to 20% of the total amount absorbed, independent of the volume of distribution. The intersubject coefficient of variation was 46% to 80% in the absorption rate constant, 27% in the distribution phase, and approximately 50% in the volume of distribution. CONCLUSIONS: PPK models adapted for enterohepatic recirculation allowed a formal assessment of the magnitude and frequency of the enterohepatic recirculation process, and the associated intersubject and intrasubject variability in healthy subjects. The PPK approach also helped to assess the correlation between the observed maximum or minimum (24 hours postdose) concentration with the model based area under the curve, confirming the appropriateness of the former measures as a surrogate of drug exposure for a possible correlation with pharmacodynamics. PMID- 11440288 TI - Migraine treatment outcomes with rizatriptan in triptan-naive patients: a naturalistic study. AB - BACKGROUND: The 5-hydroxytryptamine(1B/1D) agonists, or triptans, are the newest class of drugs to become available for the acute treatment of migraine. The class currently includes sumatriptan, zolmitriptan, naratriptan, and rizatriptan. The efficacy of rizatriptan in the acute treatment of migraine has been established against placebo and other oral triptans in controlled comparative trials. OBJECTIVE: The US Migraine Assessment Protocol (USMAP) collected data on the use of rizatriptan in a naturalistic setting reflecting clinical practice. This paper presents results for patients enrolled in the USMAP study who had never taken a triptan before the study. METHODS: At enrollment, 216 patients completed a questionnaire describing their responses to their current nontriptan medications. They were then given specially packaged samples of 4 standard 10-mg rizatriptan tablets and 4 orally disintegrating 10-mg rizatriptan tablets (wafers) and were asked to take a different formulation for each of their next 2 attacks, the sequence to be at their discretion. Within approximately 24 hours after taking rizatriptan, patients were to call a toll-free number to report their responses to rizatriptan using an interactive voice-response system. RESULTS: Within 2 hours after initial dosing of rizatriptan, significantly more patients taking either the rizatriptan tablet or the rizatriptan wafer reported onset of pain relief, had become largely symptom free, and were able to resume usual activities compared with their baseline responses to nontriptans (P < 0.05). In addition, compared with their baseline responses to nontriptans, significantly more patients taking either rizatriptan formulation had mild or no pain 2 hours after dosing (P < 0.05). More than twice as many patients taking the rizatriptan tablets or wafers were either somewhat or very satisfied with the medication compared with their satisfaction with nontriptans (P < 0.05). CONCLUSIONS: In the naturalistic setting of this study, migraineurs who had not previously taken a triptan medication reported more rapid relief of pain, more effective pain relief, and more rapid resumption of normal activities when taking rizatriptan tablets or wafers than when taking a nontriptan medication. Patients dissatisfied with their current nontriptan migraine therapy may benefit from treatment with rizatriptan. PMID- 11440289 TI - A comparison of the skin permeation of three topical 0.5% fluorouracil formulations with that of a 5% formulation. AB - BACKGROUND: Topical fluorouracil has proven efficacy in the treatment of actinic keratosis. The systemic absorption of various topical formulations of fluorouracil varies, however, and may be affected by the concentration and delivery system used. OBJECTIVE: This study compared the flux and percutaneous absorption of 3H-labeled fluorouracil from three 0.5% fluorouracil formulations incorporated into a porous microsphere delivery system with those from a commercially available 5% fluorouracil formulation. METHODS: Penetration of all formulations through the skin was sampled every 3 hours for 24 hours using full thickness human cadaver skin samples mounted in a Bronaugh flow-through diffusion cell apparatus. Total absorption was defined as the sum of the amount of cumulative flux through the skin over 24 hours and the amount retained in the skin at 24 hours. RESULTS: The flux through the skin of the 5% fluorouracil formulation was 20 to 40 times greater (normalized, 2-4 times greater) than that of the 0.5% fluorouracil formulations. A higher percentage of absorbed fluorouracil was retained in the skin after 24 hours with the 0.5% formulations (86%-92%) than with the 5% formulation (54%). CONCLUSION: These findings suggest that the 0.5% formulations are associated with less flux through the skin than the 5% formulation and, therefore, potentially less systemic exposure. PMID- 11440290 TI - A pharmacokinetic evaluation of 0.5% and 5% fluorouracil topical cream in patients with actinic keratosis. AB - BACKGROUND: Systemic absorption of topical fluorouracil, although usually low, may vary as a result of the specific skin disease, product formulation, and other factors. OBJECTIVE: The present study was conducted to determine the pharmacokinetic profile and tolerability of a new topical 0.5% fluorouracil cream formulation compared with that of a currently available topical formulation of 5% fluorouracil cream. METHODS: This was an open-label, parallel-group study in which patients with actinic keratosis (AK) were randomized to treatment with either topical 0.5% fluorouracil once daily or topical 5% fluorouracil twice daily for up to 28 days. RESULTS: Twenty-one patients (all white; mean age, 64 years) participated in the study, 11 receiving topical 0.5% fluorouracil and 10 receiving topical 5% fluorouracil. Ten patients receiving 0.5% fluorouracil and 7 patients receiving 5% fluorouracil completed the 28-day study. Plasma concentrations of fluorouracil were detectable in 3 of 10 patients treated with 0.5% fluorouracil and 9 of 10 patients treated with 5% fluorouracil; fluorouracil was detected in the urine of 5 and 9 patients, respectively. Despite the one tenth difference in drug concentration between formulations, the cumulative amount excreted in the urine of the 0.5% fluorouracil group was approximately one fortieth that of the 5% fluorouracil group. This difference may be a result of variations in vehicle formulations. At least 1 adverse event was reported by 4 of 11 patients in the 0.5% fluorouracil group and all 10 patients in the 5% fluorouracil group. The most common adverse event, facial irritation, was evident with both formulations but reached a plateau during treatment with 0.5% fluorouracil. All patients treated with 0.5% fluorouracil tolerated the full course of therapy, whereas 3 patients in the 5% fluorouracil group discontinued treatment early. No serious treatment-related adverse events were reported. CONCLUSIONS: These data suggest that 0.5% fluorouracil has minimal systemic absorption and is well tolerated in patients with AK. PMID- 11440291 TI - Cilomilast: pharmacokinetic and pharmacodynamic interactions with digoxin. AB - BACKGROUND: Cilomilast is an orally active, selective phosphodiesterase 4 inhibitor currently in clinical development for the treatment of chronic obstructive pulmonary disease. OBJECTIVE: The purpose of this study was to examine the tolerability and steady-state pharmacokinetics of cilomilast and digoxin when coadministered at standard therapeutic doses in healthy adults. METHODS: In an initial, open-label phase, healthy young adults received cilomilast 15 mg BID for 5 days. After a 7-day washout period, subjects entered a double-blind, crossover phase during which they received oral digoxin (375 microg once daily) for 2 consecutive 14-day periods with no intervening washout period. Cilomilast 15 mg BID or placebo was coadministered during the first 14-day period. Subjects then crossed over to the alternative treatment for the second 14 day period. Blood and urine samples were collected at appropriate times for evaluation of digoxin and cilomilast steady-state pharmacokinetic parameters. The size of the study was sufficient to achieve 90% power to correctly exclude an effect of cilomilast. RESULTS: Twelve of the 16 subjects enrolled completed the study. There were 4 withdrawals--1 due to noncompliance, 1 due to a positive drug screening, and 2 due to adverse events. At steady state, cilomilast 15 mg BID had no significant effect on the steady-state pharmacokinetic parameters of digoxin, with 90% CIs for both primary end points--area under the plasma concentration time curve (AUC) over a 24-hour dosing interval and minimum plasma concentration- completely contained within the specified interval for equivalence (0.80-1.25). A mean reduction in maximum observed plasma concentration of digoxin of 11% was observed during coadministration with cilomilast, and time to maximum concentration was delayed by a median of 1 hour, suggesting a small reduction in the rate of digoxin absorption. Digoxin did not appear to markedly affect cilomilast steady-state pharmacokinetics. The most frequently reported adverse event was headache. CONCLUSIONS: Cilomilast 15 mg BID had no clinically significant effect on steady-state AUC or on predose trough plasma concentrations of digoxin (375 microg once daily). The steady-state pharmacokinetics of cilomilast 15 mg BID were similar whether administered alone or with digoxin at steady state (375 microg once daily). PMID- 11440292 TI - Efficacy of single-dose intravenous dolasetron versus ondansetron in the prevention of postoperative nausea and vomiting. AB - BACKGROUND: Postoperative nausea and vomiting (PONV) is a significant problem in surgical patients. The 5-hydroxytryptamine3-receptor antagonists ondansetron, dolasetron, and granisetron are being used to prevent PONV and avoid the adverse events associated with traditional antiemetics such as antihistaminic agents, anticholinergic agents, and dopamine antagonists. OBJECTIVE: Because practitioners have taken widely differing approaches to the selection and dosing of agents in this class, this retrospective study assessed the relative efficacy of i.v. dolasetron and ondansetron in preventing PONV when used according to their approved labeling. METHODS: The medical charts of patients who underwent total abdominal hysterectomy or laparoscopic cholecystectomy and received either dolasetron 12.5 mg or ondansetron 4 mg were reviewed. Efficacy was assessed based on the number of episodes of PONV and time to the occurrence of PONV in the 24 hours after surgery. RESULTS: Of 75 medical records reviewed, 59 met the criteria for inclusion in the efficacy analysis. There were no statistically significant between-group differences in demographic or baseline clinical characteristics. The majority of patients were obese (body mass index > or = 27 kg/m2), had no history of either PONV or motion sickness, and underwent total abdominal hysterectomy. PONV occurred in 11 of 25 (44%) patients receiving dolasetron and 18 of 34 (53%) patients receiving ondansetron. Four patients receiving dolasetron experienced PONV in the first 2 hours after surgery, compared with 7 patients receiving ondansetron. CONCLUSION: There were no significant differences in efficacy between single doses of i.v. dolasetron 12.5 mg and i.v. ondansetron 4 mg in the prevention of PONV. PMID- 11440293 TI - Enhancing the benefits of pharmaceuticals: what should states do? PMID- 11440294 TI - Meta-analysis comparing newer antipsychotic drugs for the treatment of schizophrenia: evaluating the indirect approach. AB - BACKGROUND: Meta-analysis is a useful method to assess the efficacy of newer antipsychotic drugs compared with older drugs or placebo. However, few trials directly compare novel drugs to each other. OBJECTIVE: The purpose of this study was to evaluate the method of indirect meta-analysis by applying it to data on olanzapine versus haloperidol and risperidone versus haloperidol to enable a comparison between olanzapine and risperidone. METHODS: Published randomized controlled trials (RCTs) of risperidone, olanzapine, and/or haloperidol were identified through literature searches (1983 to 1999) of the MEDLINE, Current Contents, and HealthSTAR databases and reviewed. Data for the Brief Psychiatric Rating Scale (BPRS) total score, the Positive and Negative Syndrome Scale (PANSS) negative subscale, the percentage of patients using anticholinergic drugs, and the percentage of patients dropping out due to lack of efficacy, side effects, or any cause were extracted and combined using the indirect method. These findings were compared with those from a direct comparative study of olanzapine and risperidone. RESULTS: The literature search yielded 8 RCTs comparing risperidone to haloperidol and 3 comparing olanzapine to haloperidol. Only 1 trial directly comparing olanzapine and risperidone was found. In this trial, the change in BPRS total and PANSS negative subscale scores tended to be higher with olanzapine by 1.80 and 1.10, respectively, but these differences were not statistically significant. Indirect meta-analysis yielded similar results. Changes in both BPRS total scores and PANSS negative subscale scores tended to be higher with olanzapine by 0.37 and 0.54, respectively, and again, the differences were not statistically significant. In the indirect meta-analysis, the rate of anticholinergic drug use was 19.5% greater among patients treated with risperidone than among patients treated with olanzapine (P < 0.05). In the direct comparative RCT, the rate was 13.1% higher among patients treated with risperidone (P < 0.05). The dropout rates were similar for patients treated with risperidone and those treated with olanzapine in both analyses. CONCLUSION: An indirect meta-analysis of studies comparing olanzapine with haloperidol and risperidone with haloperidol yielded conclusions similar to those found in a direct comparative RCT of olanzapine and risperidone. PMID- 11440295 TI - Responsiveness of the Smoking Cessation Quality of Life (SCQoL) questionnaire. AB - BACKGROUND: The Smoking Cessation Quality of Life (SCQoL) questionnaire was developed to quantify changes in self-reported functioning and well-being associated with the smoking cessation process and to facilitate comparisons among smokers, former smokers, and nonsmokers. The SCQoL includes 5 cessation-targeted scales and the 8 multi-item scales of the Medical Outcomes Study 36-Item Short Form Health Survey. OBJECTIVE: This study was conducted to assess the responsiveness of the SCQoL by analyzing associations between SCQoL scale scores and duration of smoking abstinence. METHODS: The SCQoL was administered at a screening visit and 2 to 6 weeks after screening as part of a longitudinal study. Study participants included smokers who intended to quit smoking. Subjects were required to purchase nicotine inhalers and were prompted to quit smoking before follow-up. Based on self-reported duration of abstinence at follow-up, subjects were categorized as recidivists (0 days smoke free), short-term abstainers (1-7 days smoke free), or longer-term abstainers (>7 days smoke free). Kruskal-Wallis tests were used to compare changes in scale scores from screening to follow-up among the 3 groups. RESULTS: The internal consistency reliability of the 13 SCQoL scales ranged from 0.67 to 0.92. Subjects who maintained abstinence for longer periods experienced smaller declines in health-related quality of life between the screening and follow-up assessments. Differences among the 3 groups were driven primarily by differences between recidivists and longer-term abstainers. CONCLUSIONS: The results are consistent with research indicating that recidivists report a greater number of (and more severe) cessation-related symptoms than abstainers. The findings of this investigation provide evidence for the responsiveness of the SCQoL. PMID- 11440296 TI - The effects of postal and telephone reminders on compliance with pravastatin therapy in a national registry: results of the first myocardial infarction risk reduction program. AB - BACKGROUND: Noncompliance with cardiovascular therapy and prevention initiatives is well documented. OBJECTIVES: The purpose of the First Myocardial Infarction (MI) Risk Reduction Program, an open-label drug registry involving mainly primary care patients at increased risk of a first MI, was to examine the effects of postal and telephone reminders, as well as demographic and other baseline characteristics, on patient self-reported compliance with pravastatin treatment. A second objective was to determine whether regimen adherence was associated with the adoption of other lifestyle modifications recommended to decrease the risk of coronary artery disease. METHODS: Patients with risk scores of > or = 4 on a scale of -1 to +16 for men and -1 to +17 for women on the First Heart Attack Risk Test were considered to be at increased risk of a first MI and eligible for enrollment in the registry program. An elevated total cholesterol level despite dietary interventions was an additional inclusion criterion. Patients were prospectively randomized (4:1) to either an intervention involving postal and telephone reminders (about coronary risk reduction and medication compliance), which were sent during the first 2 months of pravastatin treatment, or usual care. Both groups received reminder postcards at 4 and 5 months, in addition to counseling by physicians about coronary risk reduction. At 3 and 6 months (or study discontinuation), patients completed and mailed to the program-coordinating center questionnaires concerning compliance with care, including current use of prescribed pravastatin, as well as self-reported adoption of other lifestyle modifications, such as changing eating habits, losing weight, increasing physical activity, and/or quitting smoking. Compliance with pravastatin therapy and with these coronary risk-reducing behaviors was also assessed by physicians at the 3 month follow-up visit. RESULTS: A total of 10,335 patients were in the intervention group, and 2765 received usual care. The 2 groups were well balanced at baseline with respect to age, race, and total cholesterol values. Neither early reminders nor baseline patient characteristics were significantly associated with reported pravastatin compliance rates, which were approximately 79% overall. However, according to self-reports at 6 months, regimen compliance was associated with the adoption of other coronary risk-reducing behaviors. CONCLUSIONS: The results of this study suggest that early telephone and postal reminders do not improve compliance with drug treatment or with recommended coronary risk-reducing behaviors. PMID- 11440297 TI - Application of an optical immersion-gel in a flow cytometer with horizontally oriented objective. AB - In certain flow cytometry systems, it is desirable to use immersion optics to obtain optimum fluorescence yield. This is important when propidium iodide and other DNA fluorochromes are used that have weaker fluorescence emission compared to DAPI, when a lamp is used instead of a laser and when the DNA concentrations are low. Our Partec PA II with a horizontally oriented objective and a vertically oriented flow chamber precludes using a liquid immersion medium. The problem was solved using an optical gel with appropriate characteristics. This gel is commercially available and commonly used for connecting glass fiber cables, but has never been used for microscopy before. Compared to the manufacturer's objective (40 x, aperture 0.8), the fluorescence yield was improved approximately four-fold using the optical gel and a 40 x glycerol objective (aperture 1.25). This innovation widens the applicability of flow cytometers with horizontally oriented objectives and vertical flow chambers. We expect it to facilitate the use of propidium iodide as a DNA stain, especially when interspecific genome size comparisons are to be done and base ratio dependent bias must be avoided. PMID- 11440298 TI - Gram staining and lectin binding properties of Myxosporea and Sporozoea. AB - The staining method developed by Christian Gram was introduced as a simple and highly selective tool for demonstrating myxosporean and coccidian sporogonic stages. When using standard blood staining procedures for those enigmatic parasites it is sometimes difficult to distinguish them from fish host tissue. They clearly exhibit a partial gram-positive reaction in histological sections, but staining is variable in air dried fish organ imprints. To visualize the gram negative background of different host tissue components in histological sections, the conventional safranin counterstain of the gram protocol may be modified as follows: after application of 2% crystal violet (basic violet 3) and Lugol's solution, sections are stained with 0.1% nuclear fast red-5% aluminum sulfate and 0.35% aniline blue (acid blue 22) dissolved in saturated aqueous picric acid. Replacement of the gram-specific dye crystal violet with 2% malachite green gave similar results in organ imprints containing myxospores or coccidia, but only in sections containing myxosporea. Staining for 1 min with an aqueous solution of 0.5% malachite green and followed 1 min washing was sufficient for rapidly demonstrating the parasite spores in organ imprints of both myxosores and oocysts. With regard to the role of acid mucopolysaccharides and other carbohydrates in the gram reaction of spores, alcian blue 8GX staining was compared to the binding of FITC-labeled WGA, GS I and GS II. Each lectin was applied at 20 microl/ml PBS, HEPES for 1 hr. Whereas WGA yielded a nonspecific pattern like the alcian blue staining, GS II resulted in a pattern similar to the gram staining results. This binding was weak in untreated specimens, but was significantly enhanced when digested first within trypsin overnight in a humid chamber at 37 degrees C. The binding of GS II to both myxosporidian and coccidian spores suggests that they are both composed of polymers containing N-acetyl-D glucosamine residues. Furthermore, the results suggest that this hexosamine plays a key role in the gram reaction. PMID- 11440299 TI - Comparison of historic Grubler dyes with modern counterparts. AB - Seventeen Grubler dyes produced in Germany between 1880 and 1939 were examined in this study. These dyes were: fuchsin-bacillus, diamond fuchsin, fuchsin S acid, rubin S, safranin O water soluble, safranin yellowish water soluble, methyl eosin, Sudan III, scarlet R, auramine, orange G, aniline blue, pyronin, carmine, lithium carmine, hematein and aurantia. Spectrophotometry and staining characteristics were used to determine the maximum absorbance and efficacy of each dye in common staining techniques. The spectral curves and staining characteristics of these dyes compared well with modern dyes used as controls. Fuchsin bacillus and diamond fuchsin are synonyms for basic fuchsin. Fuchsin S acid and rubin S are synonyms for acid fuchsin. The scarlet R sample was the same as the Sudan III. The two safranins were the same. The basic fuchsin samples were unsuitable for preparation of Schiff's reagent. Both basic fuchsin and pyronin samples were less concentrated than modern counterparts. It is noteworthy that the dyes worked well after up to 100 years in storage, and this observation indicates that dyes can have a long shelf life when stored in cool, dry, air tight conditions. PMID- 11440300 TI - Rapid plastic embedding is compatible with colorimetric detection following whole mount in situ hybridization in plant specimens. AB - In performing in situ hybridizations, nonisotopic nucleic acid labeling coupled with colorimetric detection offers a safer, easier and more rapid alternative to using radioactively labeled nucleic acid probes and microscopic autoradiography. Whole mount in situ hybridization is also advantageous, because many samples can be processed identically and the reduced handling of specimens greatly reduces the risk of exposing tissues to RNase(s). The thickness of whole mount specimens, however, often prevents accurate determination of sites of expression within specific tissues. Although post-hybridization embedding and sectioning is a solution to this problem, the precipitate formed following the common colorimetric detection procedure is soluble in the organic solvents used for dehydration prior to embedding. We have developed a dehydration and embedding procedure that takes advantage of the compatibility of L.R. White resin containing 10% (v/v) polyethylene glycol 400, and heat polymerized. The addition of the plasticizer allows L.R. White embedded tissues to be sectioned at 10 microm providing excellent signal contrast. PMID- 11440301 TI - A stain for demonstrating Helicobacter pylori in gastric biopsies. AB - Helicobacter pylori continues to be a significant health care problem. It is associated with a variety of stomach disorders such as gastritis, gastric ulcer disease, gastric carcinoma and B-cell gastric lymphoma. One common method diagnosing an infection by this bacterium is microscopic examination of routine processed gastric or duodenal biopsies. With this type of specimen, it is necessary to demonstrate visually the presence of H. pylori using an appropriate staining technique. This paper presents a simple staining technique for demonstrating H. pylori in gastric biopsy specimens using carbol fuchsin staining against a contrasting background of light green. PMID- 11440302 TI - Classification and counting of fluorescent pollen using an image analysis system. AB - Pollen viability is commonly assessed by fluorochromatic reaction (FCR) because of the high correlation between positive fluorescence of the pollen grains and their ability to germinate. One of the advantages of this method is its simplicity. An experiment to test FCR analysis for reproducibility, however, showed that results are affected by subjectivity. There is little consistency between analysts, and assessment by the same analyst may differ for the same pollen sample image examined at different times. These problems were solved by a computerized image analysis system that provides a method for classifying positive and negative fluorescent pollen and automatic counting of the grains in each class. The computerized image analysis system does not change the biochemistry of the FCR test, but avoids some experimental errors owing to the subjectivity of the analyst. Microscope images of the pollen after FCR were digitized and later analyzed by specially designed software, "Plant Meter." This software deletes the dark background of the image to isolate the grains, and subsequently counts positive and negative fluorescent pollen grains. An experiment was carried out to validate software output and it showed reliable results. Moreover, the software is user friendly and very little training is necessary for analysts to achieve reliable results. PMID- 11440303 TI - A simple, rapid method for measuring the percentage of water in alcohols used for dehydrating tissues. AB - A method is described for determining the concentration of water in alcohols that have been used for dehydrating specimens. The alcohol to be tested is added dropwise to 5.0 ml of xylene, counting the number of drops that causes turbidity, then the number of additional drops required to make the liquid transparent again. The amount of alcohol required to clear the turbidity increases steeply with water content, providing an easy and reliable assay method for water content of 90% to 97% alcohol baths. Alcohol containing 2% or less water does not become turbid when xylene is added. PMID- 11440304 TI - Preservation of the morphological and molecular stability of embryonic tissues. AB - There is presently great interest in using early embryonic tissues, particularly human tissue, for studies of protein and gene expression. Embryonic human tissue is very fragile, and delays often occur before it can be properly prepared for scientific study. Using chick embryos, we have studied the effects of delaying fixation or biochemical isolation on the preservation of cytological characteristics and biochemical molecules. Our study shows that by 60 min post harvest, tissue morphology and immunofluorescence staining degrades, but the total mRNA profile remains stable. This study suggests that the time between removal of the tissue and fixation is critical to the results and that the critical time is much shorter for embryonic tissues than for more developed tissues. Our results have implications for all research where embryonic tissues are harvested but not processed immediately. PMID- 11440305 TI - Effects of time and temperature during attachment of sections to microscope slides on immunohistochemical detection of antigens. AB - To study the effects of time and temperature on attachment of tissue sections to microscope slides, we examined the intensity of immunohistochemical staining of selected antigens in nine different neoplastic and normal tissues after attaching sections at different times and temperatures. Typically, both the temperature and time are minimized when tissue sections attached to slides; however, suboptimal times and temperatures during attachment may result in either loss of tissue due to poor attachment or the necessity for inconvenient staining regimens. Using standard immunohistochemical techniques, 5 microm tissue sections were attached at 58 degrees C for 1, 4 and 24 hr. In a separate study, 5 microm tissue sections were attached for 16 hr at 58, 68 and 80 degrees C. The intensity of staining decreased slightly when the tissue sections were heated at 80 degrees C for 16 hr, but there was little or no decrease when tissues were heated at 68 degrees C or lower for 16 hr, or at 58 degrees C for up to 24 hr. PMID- 11440306 TI - A new technique for studying the stylet tracks of homopteran insects in hand sectioned plant tissue using light or epifluorescence microscopy. AB - Homopteran insects, such as aphids, psyllids and scales, inject a proteinaceous salivary sheath into their host plant tissue during feeding. This sheath, also referred to as a stylet track, remains in the tissue after the stylets are withdrawn, and is useful for studying plant resistance to insects and plant virus transmission. We describe a new method for studying stylet tracks. Hand microtome sectioned plant material was fixed and cleared in ethanol. The stylet tracks were stained with acid fuchsin and counterstained with aniline blue or fast green. The acid fuchsin stained stylet tracks were pink to red under light microscopy, and orange under TRITC epifluorescence. Stylet tracks in unstained sections autofluoresced under DAPI epifluorescence. This new technique is significantly faster and less complex than previous techniques, and permitted visualization of stylet tracks with light or epifluorescence microscopy within 1 hr of collecting fresh plant material. The technique was also applicable to a broad range of homopterans and plant taxa and provided excellent photomicrographs. PMID- 11440307 TI - Modified silver staining of nucleolar organizer regions to improve the accuracy of image analysis. AB - Silver staining of nucleolar organizer regions (NORs) and their subsequent quantification by image analysis are used increasingly in human pathological specimens and experimental models. Because certain conditions determined by the type of tissue and/or its fixation render AgNOR segmentation for image analysis difficult due to insufficient contrast or nonspecific silver precipitation, we propose three improvements to the original technique to overcome these difficulties. Pretreatment with 7% nitric acid produced very distinct dark brown images of AgNORs on a yellow background. The gradient of background colors allowed easy discrimination of nucleolar, nuclear and cytoplasmic structures. Seven morphometric parameters related to number, size and shape of AgNORs were evaluated quantitatively by image analysis on sections pretreated with nitric acid and on adjacent sections treated with citrate buffer in a wet autoclave according to the most widely accepted method for image analysis of AgNOR. Both methods yielded similar results. A second improvement was achieved by coating the slides with 7% celloidin solution in ethyl alcohol-ether prior to AgNOR staining and acid pretreatment. This coating prevented nonspecific silver deposition on argyrophilic bacteria and other tissue debris in human vaginal smears that could make visualizing AgNOR sites difficult. Finally, placing sections face down on the staining solution prevents the formation of nonspecific silver precipitates. These procedures can be applied together or separately according to the requirements of the material to be evaluated. PMID- 11440308 TI - Application and modification of in situ RT-PCR for detection and cellular localization of PAC1-R splice variant mRnas in frozen brain sections. AB - Many important biopolymers such as neurotransmitters, modulators, transporters and receptors are expressed in discrete regions of the brain or other tissues, and they often occur at extremely low concentrations; therefore, a sensitive detection system is required to map their distribution. To study the precise distribution patterns of the splice variants of the PAC1 receptor, which specifically binds pituitary adenylate cyclase-activating polypeptide (PACAP) with affinity in the nano- or picomolar range, we have applied an in situ reverse transcription-polymerase chain reaction (RT-PCR) technique in frozen tissue sections. We describe here a modified protocol using a single rTth enzyme, which can synthesize cDNA from RNA, then PCR amplifying it in a single reaction mixture by varying the times and temperatures of a thermal cycler. The primer pairs were the same as those used in the solution phase RT-PCR that had been used to obtain the expected bands of the amplified products previously. A nonradioactive labeling system with digoxigenin conjugated with peroxidase or fluorescence for signal detection was compared. The gene expression of two PAC1-R splice variants in the rat motor nucleus is first reported here. PMID- 11440309 TI - Immunohistochemistry combined with periodic acid-Schiff for bovine mammary gland with protothecal mastitis. AB - Immunohistochemistry (IHC) of bovine cytokeratin combined with periodic acid Schiff (PAS) was applied to study the pathogenesis, localization and distribution of Prototheca zopfii in bovine mammary protothecosis. The standard immunohistochemical procedure using anti-bovine cytokeratin was employed before and after PAS staining to optimize this combined method. The best results were obtained when IHC procedures were performed first. Most of the epithelial cells reacted strongly with the pancytokeratin antibody. Protothecal cell walls stained well with PAS. Algal organisms were present within the lumen and between the epithelial lining and basement membrane of the affected alveoli, but not inside the positive mammary epithelial cells. This combined staining method resulted in clear alveolar epithelial detail and good contrast between the epithelial cells and algae, and contributed to studying the pathogenesis of P. zopfii in mammary protothecosis. PMID- 11440310 TI - Histological techniques and microscopic analysis of biological agents for preservation of human bone remains. AB - Microscopy and histology techniques can be applied to morphological study of fungi and bacteria contaminating ancient human osteological remains. Undecalcified samples are cut with a diamond rotary blade microtome and an original technique was applied using adhesive tape to prevent damage to the bone surface during sectioning. We used light microscopy, polarized light microscopy, and epi-illumination fluorescence systems. Common dyes can be applied to 80 microm sections using classic staining techniques to reveal the architecture of bone and the presence of infecting biological agents. PMID- 11440311 TI - Application of triple immunohistochemistry to characterize amyloid plaque associated inflammation in brains with Alzheimer's disease. AB - Inflammation, characterized by the presence of activated microglia and reactive astrocytes (gliosis), has been described in Alzheimer's disease (AD). We used our routine single immunohistochemical (IHC) labeling protocol to label amyloid plaques, an AD neuropathological hallmark, activated microglia, and reactive astrocytes in serial sections of AD hippocampus and entorhinal cortex of brain. Although most amyloid plaques were associated with inflammation throughout the serial sections, the extent of microglial and astrocytic activation varied among the amyloid plaques. We also observed a population of amyloid plaques that did not appear to coincide with immunolabeled microglia and astrocytes in serial sections, leading us to speculate that some amyloid plaques are not associated with inflammation. Because serial sectioning limited our ability to confirm these findings, we developed a triple IHC protocol to investigate the association of activated microglia and reactive astrocytes simultaneously with amyloid plaques in sections of AD brain entorhinal cortex and hippocampus. Unlike the potential errors of extrapolating descriptive information from routine IHC or histochemical staining methods on sectioned tissues, triple IHC allowed direct characterization of three differently colored antigens in situ. The success of the protocol depended on selection of distinguishable color schemes and resolution of other critical technical elements including the compatibility of the reagents and the sensitivity and sequence of the detection systems. The results of the triple IHC protocol clarified the spatial relation of microglia and astrocytes with amyloid plaques and provoked novel interpretations about the roles of inflammation in AD brain tissues. We categorized three distinct populations of amyloid plaques related to of inflammation: 1) Abeta42 immunoreactive (a marker of amyloid plaques) amyloid plaques without activated microglia or reactive astrocytes, 2) Abeta42-positive amyloid plaques with HLA-DR (a marker of microglia)-positive microglia and no astrocytes, 3) Abeta42-positive amyloid plaques among HLA-DR and GFAP (a marker of astrocytes) immunoreactive astrocytes. Most amyloid plaques had varying degrees of activated microglia and reactive astrocytes. Some of the amyloid plaques were not associated with inflammation while others were associated only with activated microglia. These findings suggest that amyloid plaques without associated inflammation may represent recently formed plaques and that the presence of amyloid plaques in AD brains may activate microglia prior to gliosis. Furthermore, the shape of the amyloid plaques may be altered subsequently from its typical spherical to an aspherical shape by the inflammatory cells. PMID- 11440312 TI - Field measurements of electromagnetic interference in hearing aids. AB - This investigation was a preliminary field study to determine the acoustic and perceptual characteristics of hearing aid distortion generated by digital wireless telephones, the usability of the telephones under field conditions, and the extent of bystander interference under field conditions. A two-channel analog to-digital converter was used to monitor voltages generated by an acoustic (real ear) and electromagnetic probe. Digital recordings of interference and speech plus interference were made on a laptop computer. Fifty-three hearing aid wearers listened to interference and speech plus interference through personal communication service 1900 and time division multiple access digital wireless telephones and rated them in terms of annoyance experienced and usability of the wireless telephone. Ratings of annoyance were also done for the bystander condition. Approximately 80 percent of the sample rated the telephones as unusable; on the other hand, 70 to 90 percent experienced no annoying interference from telephones being used by another person seated nearby (bystander condition). PMID- 11440313 TI - Suitcase lab measurement of digital cellphone interference levels on hearing aids. AB - A low-cost, "real-life" method for measuring the interference caused by digital wireless (cell-phones) telephones in hearing aids is proposed. Data would be valid for specific telephone and hearing aid models. The estimated equipment cost is $500. PMID- 11440314 TI - Wireless telephones and hearing aids: an overview. AB - Wireless telephones are becoming increasingly common and will be more so in the future. Unfortunately, many are unusable with hearing aids and may actively interfere with the normal use of hearing aids because of the electromagnetic interference (EMI) generated by wireless telephones. The EMI generated by digital wireless telephones is much greater than that of analog wireless telephones. More research and design work needs to be done before wireless telephones can be considered accessible to people who use hearing aids. PMID- 11440315 TI - Practical information for audiologists on access to wireless telephones. AB - Analog cellular telephone service is being phased out in favor of digital wireless services, which are less accessible to people with hearing loss. As a result, audiologists can expect to receive an increasing number of inquiries from clients about using wireless telephones with hearing aids and other assistive technologies. In this article, the underlying transmission technology, telephone handset, roaming, and current solutions to accessibility problems are described. Public policy affecting the accessibility of wireless telephones to people with hearing loss is explained. Alternatives to wireless voice services are explored, and resources for information in a quickly changing industry are provided. PMID- 11440316 TI - The European experience. AB - This article presents an overview of past and current experiences with time division multiple assess-based (Global System for Mobil Communication) mobile telephones in Europe as seen by the European Hearing Instrument Manufacturers Association. Initial fear of widespread interference problems for hearing aid users in general owing to use of a new generation of mobile telephones seems unjustified. The background for the International Electrotechnical Commission 118 13 standard for measuring interference is described. No solution to complete elimination of interference problems resulting from direct contact between hearing aids and mobile telephones has yet been found. Several reports on the subjects are cited, and new work on measurement standards for near-field situations is mentioned. PMID- 11440317 TI - Digital wireless telephones and hearing aids: new challenges for audiology. PMID- 11440318 TI - Wireless telephone-hearing aid electromagnetic compatibility research at the University of Oklahoma. AB - A multiphase study examining electromagnetic compatibility (EMC) between wireless digital telephones and hearing aids has been under way at the University of Oklahoma EMC Center since May 1995. In a phase 1 clinical study involving 68 hearing aid wearers, interference varied significantly by telephone technology, hearing aid type, and hearing loss characteristics. More than 80 percent of the tests resulted in either no interference or a detection threshold distance less than 1 meter. Metallic shielding of the units yielded positive results. Various elements of phase 2 involved instrument-based tests of hearing aid interference using telephones in a sound-isolation chamber and radio frequency signals in a waveguide, along with clinical studies of speech-to-interference ratios, all leading to the development of standards of measurement and performance criteria for telephone emissions and hearing aid immunity. Results to date confirm that bystander interference is of less concern than user interference, which is the focus of continuing research. PMID- 11440319 TI - Hearing aid and cellular telephone compatibility: working toward solutions. AB - The ability of hearing aid wearers to use cellular telephones using digital transmission formats has become a prominent issue. This article presents the requirements, technical challenges, and emerging standards for the combined use of hearing aids with cellular telephones. PMID- 11440320 TI - The Australian experience: global system for mobile communications wireless telephones and hearing aids. AB - A digital, wireless telephone system, the Global System for Mobile Communications (GSM), was introduced in Australia in 1993. Studies were conducted at the National Acoustic Laboratories to determine how the use of GSM mobile telephones (i.e., wireless telephones) causes interference in hearing aids and how hearing aids could be made more immune to such interference. A measurement system was developed, and measurements are presented for a variety of standard and treated hearing aids. Effective treatments include the addition of shunt capacitors to the hearing aid circuit and shielding by applying a conductive coating to the hearing aid case. Some recently produced microphones also increase immunity. High immunity levels can be achieved in future hearing aids, sufficient to prevent interference from other people's use of a wireless telephone and that often permit use of digital mobile telephones by hearing aid users. Audiologists will have an increasing role in advising hearing aid wearers on the use of wireless telephones or other communications equipment. PMID- 11440321 TI - The nature of electromagnetic interference. AB - This report provides a brief tutorial on the underlying physical forces that lead to interference with hearing aids and assistive listening devices, as well as measurement issues and possible solutions to the problem. PMID- 11440322 TI - Choosing our view of osteoporosis. PMID- 11440323 TI - Highlights of the Fourth Annual Conference on Osteoporosis, Amelia Island, Florida, February 22-24, 2001. PMID- 11440324 TI - NIH Consensus Development Panel on Osteoporosis Prevention, Diagnosis, and Therapy, March 7-29, 2000: highlights of the conference. PMID- 11440325 TI - Medication update. PMID- 11440326 TI - Distinguishing between clinical guidelines and covered services. PMID- 11440327 TI - Outcome study of surgical treatment for axial neck pain. AB - BACKGROUND: We reviewed our surgical treatment of chronic axial cervical pain over a 4-year period to determine whether surgery in selected cases was associated with favorable outcomes. METHODS: We retrospectively studied 27 consecutive cases (20 patients with follow-up) of longstanding axial cervical spine pain treated surgically by a single surgeon from June 1994 through August 1998. Diagnostic workup included the following when appropriate: Minnesota Multiphasic Personality Inventory (MMPI) with interview, provocative diskography (with a nonpainful control level), single photon emission computed tomography (SPECT), and diagnostic facet injection. Twenty patients (74%) responded to a postoperative telephone survey. RESULTS: For general outcome measures, 85% of patients reported satisfaction with pain relief and surgical result. Ninety-five percent stated they would repeat the procedure; 85% manifested improvement in Prolo score. CONCLUSIONS: Surgical treatment of chronic axial neck pain, when preceded by thorough evaluation, can yield excellent clinical results. PMID- 11440328 TI - Eye symptoms and use of eye protection among seasonal and migrant farmworkers. AB - BACKGROUND: Farmworkers face an array of environmental and workplace hazards that pose risks of eye damage from accidents involving agricultural chemicals and equipment and from exposure to wind, dust, and ultraviolet rays. Eye safety risks are known among industrial and agricultural occupations, yet little is known about the prevalence of eye symptoms and use of eye protection among farmworkers. METHODS: A survey of 197 Latino farmworkers in North Carolina used self-report data to measure workers' prevalence of eye symptoms and eye protection use. RESULTS: Eye pain and redness after working all day in the field were reported by more than 40% of workers. Most (98.4%) reported not wearing sunglasses when working in the fields; reasons included lack of sunglasses and interference with field tasks. CONCLUSIONS: Eye symptoms are prevalent in this population. Failure to use eye protection indicates the need for further education and training of Latino farmworkers and their employers about occupational risks to eyesight. PMID- 11440329 TI - Use of oral creatine as an ergogenic aid for increased sports performance: perceptions of adolescent athletes. AB - BACKGROUND: Competitive athletes, including adolescents, seek ways to gain advantage over competitors. One ergogenic aid is creatine, a naturally occurring nitrogen compound found primarily in skeletal muscle. Increasing creatine levels may prolong skeletal muscle activity, enhancing work output. METHODS: A questionnaire assessing awareness and use of creatine supplementation was completed by 674 athletes from 11 high schools. Data were statistically analyzed to determine variation among groups. RESULTS: Of those surveyed, 75% had knowledge of creatine supplements, and 16% used creatine to enhance athletic performance. Percentage of use increased with age and grade level. Awareness and use were greater among boys than girls. Adverse effects were reported by 26%. Most athletes consumed creatine using a method inconsistent with scientific recommendations. CONCLUSIONS: Use of creatine by adolescent athletes is significant and inconsistent with optimal dosing. Physicians, athletic trainers, and coaches should disseminate proper information and advise these adolescent athletes. PMID- 11440330 TI - Stroke awareness among Georgia adults: epidemiology and considerations regarding measurement. AB - BACKGROUND: To design and evaluate interventions for reducing the impact of stroke in Georgia, we assessed knowledge of signs, risk factors, and burden of stroke. METHODS: Adults in Georgia were studied with a random digit dial telephone survey. RESULTS: Answering an unaided question, 39% of 602 respondents named > or =1 stroke warning sign. Awareness was considerably greater when assessed with prompted questions. Most respondents (70%) said they would call 911 if someone had a stroke; almost all (95%) considered stroke an emergency. Risk factor awareness ranged from 97% (previous stroke) to 69% (diabetes). Altogether, 6% reported having had a stroke; 48% reported a stroke in their family. CONCLUSIONS: Georgia adults have low awareness of stroke warning signs. Our findings underscore the importance of conducting an effective educational campaign. Furthermore, a need exists for questions on stroke awareness that approximate more closely the situation in which a person must identify a potential stroke. PMID- 11440331 TI - Changing antibiotic sensitivity patterns at a university hospital, 1992 through 1999. AB - BACKGROUND: The emergence of antibiotic-resistant organisms is of great concern in the medical community. Antibiotic sensitivity patterns were studied at a large university hospital. METHODS: From 1992 through 1999, susceptibility testing was done and results recorded for all isolates of Streptococcus pneumoniae, Enterococcus sp, Staphylococcus aureus, coagulase-negative Staphylococcus, Eshcherichia coli, Haemophilus influenzae, and Pseudomonas aeruginosa. Microbiologic and sensitivity data were reviewed and compiled. RESULTS: Over the 8-year period, several common bacterial pathogens declined in susceptibility to various antimicrobial agents. Most notable were the decreased sensitivities of S pneumoniae to penicillin (96% to 63%), coagulase-negative Staphylococcus to oxacillin (50% to 38%), and P aeruginosa to aminoglycosides [(gentamicin (85% to 64%), tobramycin (96% to 83%), amikacin (92% to 74%)] and ciprofloxacin (85% to 69%). CONCLUSIONS: These decreased antibiotic sensitivities reflect increased bacterial selection pressure as a result of widespread antibiotic use. A combined approach involving infection-control specialists, infectious disease physicians, and hospital administrators is necessary to address this increasingly difficult problem. PMID- 11440332 TI - Tobacco use and skin disease. AB - BACKGROUND: The primary objective of this review is to evaluate the mucocutaneous manifestations of tobacco use. METHODS: Computerized literature searches were conducted for English language articles related to skin/mucous membrane disease and use of tobacco. The primary criterion for assessing data quality and validity was the demonstration of a causal relationship between tobacco use and skin/mucous membrane disease. RESULTS: This review of the literature shows that a number of disorders and diseases of the skin and mucous membranes are related to tobacco use. CONCLUSIONS: Since millions of persons use tobacco despite its well publicized relationship to increased mortality, knowledge of the mucocutaneous morbidity associated with tobacco use may help physicians in counseling their patients. PMID- 11440333 TI - Pulmonary hypertension associated with HIV infection. AB - Pulmonary hypertension occurs with increased frequency among patients with human immunodeficiency virus (HIV) infection. Although the pathogenesis of HIV associated pulmonary hypertension remains unknown, it appears to occur independently of other risk factors associated with pulmonary vasculopathy, such as chronic hepatitis C infection and intravenous drug use. Signs and symptoms are typical of those immunocompetent patients with primary pulmonary hypertension, but because many HIV-infected patients are receiving intensive medical supervision, the diagnosis of pulmonary hypertension is often made at an earlier stage. Acute responses to epoprostenol are similar to those among non-HIV infected individuals, but the benefits of long-term, intravenous treatment with epoprostenol in HIV-infected patients is unknown. Future investigations should define the true incidence of pulmonary hypertension and the long-term effects of epoprostenol on survival among HIV-infected individuals. Physicians should be alert to possible pulmonary hypertension in persons infected with HIV. PMID- 11440334 TI - Splenic angiosarcoma and iron deficiency anemia in a 43-year-old man. AB - Thrombocytopenia and microcytic anemia are two laboratory findings that alone or together suggest an underlying disease process. Both are found throughout particular age groups and have broad differential diagnoses. Angiosarcomas are rare neoplasms from the lining of blood vessels. Primary splenic angiosarcoma is an even rarer neoplasm, first reported in the late 1870s. We report a case of primary splenic angiosarcoma in a patient with thrombocytopenia, microcytic anemia, and splenomegaly. PMID- 11440335 TI - Multiple penetrating injuries to the heart diagnosed with ultrasonography. AB - Penetrating heart injury poses significant diagnostic and therapeutic challenges. Patients may initially appear in extremis or in stable condition. Surgeon performed ultrasonography is effective in determining the presence or absence of tamponade. Definitive repair requires cardiorrhaphy, using a median sternotomy or thoracotomy incision, depending on the suspected injury site(s). Frequent postoperative complications can usually be managed without reoperation. Echocardiography, electrocardiography, and cardiac catheterization may be used postoperatively in reassessment. A patient with stab wounds to both ventricles of the heart is described. PMID- 11440336 TI - Where have all the hours gone? PMID- 11440337 TI - Choreiform movements in dialysis patient taking valacyclovir and famciclovir. PMID- 11440338 TI - Drug-facilitated sexual assault. PMID- 11440339 TI - Generalized epileptic disorders: an update. PMID- 11440340 TI - Anticonvulsant efficacy of gabapentin on kindling in the immature brain. AB - The anticonvulsant and motor effects of gabapentin (GBP) were evaluated in rat pups aged 16-17 days. Fourteen-day-old rat pups received an implanted stimulating electrode in the amygdala unilaterally. Kindled seizures were produced on day 16 of life by repeatedly applying an electrical current stimulus to the amygdala electrode. Animals received kindling stimulation until they achieved three consecutive generalized convulsions. On day 17, rat pups received one of four doses of GBP 10, 25, 50, or 100 mg/kg. After receiving GBP, rat pups again received electrical stimulation to the amygdala electrode to determine the extent to which GBP prevented the kindled seizure. Anticonvulsant effects were found at doses as low as 10 mg/kg. A separate group of naive rats received GBP to determine the motor effects of each treatment dose. Impaired motor performance, quantified as time on a balance beam, occurred at doses of >or=50 mg/kg. In summary, our data indicate that in immature rats, GBP exerts an anticonvulsant effect against kindled seizures at doses that do not significantly impair motor performance. PMID- 11440341 TI - Newly diagnosed unprovoked epileptic seizures: presentation at diagnosis in CAROLE study. Coordination Active du Reseau Observatoire Longitudinal de l' Epilepsie. AB - PURPOSE: We describe first unprovoked seizures and newly diagnosed epilepsies at initial presentation, with a special emphasis on epilepsy syndromes, in a large cohort recruited in the mid-1990s in France. METHODS: The French Foundation for Research on Epilepsy set up a network to conduct a prospective study of patients with newly diagnosed unprovoked seizures. Information was provided by 243 child or adult neurologists. Four neurologists classified each case according to the International League Against Epilepsy (ILAE) criteria. First-seizure patients and patients with previously undiagnosed seizures were compared. RESULTS: Between May 1, 1995, and June 30, 1996, 1,942 patients aged from 1 month to 95 years were identified: 926 (47.7%) with a single seizure and 1,016 (52.3%) with newly diagnosed epilepsy. All but 17 patients had EEGs. In the first-seizure and newly diagnosed-epilepsy groups, neuroimaging studies were performed in 78.2 and 68.3% of patients, and medication prescribed in 54.1 and 89.6%, respectively. There were significant differences between the two groups with respect to age at onset and diagnosis, sex, etiology, several specific syndromes, as well as the type and presentation of initial seizure. In patients for whom the first seizure was convulsive, only sex, multiple seizures in a day or status epilepticus, and cryptogenic localization-related syndrome differed between the two groups. CONCLUSIONS: Approximately half of patients who first came to attention for an unprovoked seizure already met epidemiologic criteria for epilepsy. There were significant differences between the types of patients with a first seizure and those with newly diagnosed epilepsy. One or several seizures at diagnosis did not influence the diagnostic assessment of the patients but had a strong influence on the initiation of treatment. PMID- 11440342 TI - Classification of epileptic seizures: a comparison of two systems. AB - PURPOSE: Extensive experience with video-EEG seizure monitoring, notably in the setting of epilepsy surgery programs, has exposed the limited value of the current International Classification of Epileptic Seizures (ICES) for providing relevant localizing information. To overcome this limitation, a Semiologic Seizure Classification (SSC) has recently been proposed. This study aimed to assess and to compare the usefulness and reliability of both systems in the setting of a tertiary epilepsy center. METHODS: Three epileptologists independently reviewed video-taped seizures, randomly selected from the archive of the Epilepsy Monitoring Unit. They were blinded to the EEG findings and final diagnosis and classified all seizures according to both classifications. RESULTS: One hundred thirty-eight seizures from 60 patients (age range, 2-59 years) were reviewed (maximum, three seizures per patient). Fifty-five seizures from 20 patients were recorded in the setting of presurgical evaluations, and the remainder as part of regular diagnostic evaluations. The average interobserver agreement was higher for SSC (63.3%, kappa = 0. 56) than for ICES (38.6%, kappa = 0.41). Some categories of SSC, such as hypermotor or automotor, had the best interobserver agreement, and were strongly correlated with the anatomic localization of the seizures (frontal and temporal lobe, respectively). All reviewers agreed that SSC provided a better description of the seizures than did ICES, in 60% of the patients. CONCLUSIONS: SSC provides a more comprehensive picture of epileptic seizures than does ICES, notably in patients with localized epilepsy syndromes, and appears to be very useful and reliable, particularly in the setting of specialized epilepsy centers. PMID- 11440343 TI - Dipole localization for identification of neuronal generators in independent neighboring interictal EEG spike foci. AB - PURPOSE: We evaluated dipole localizations of independent neighboring interictal spike foci using scalp electroencephalogram (EEG) to identify neuronal generators of epileptic discharges. METHODS: Three pediatric patients with extratemporal lobe epilepsy who had two independent neighboring interictal spike foci on scalp EEG were studied. Prolonged video EEG was digitally recorded from 19 scalp electrodes, whose positions were registered using a three-dimensional digitizer. Interictal spikes were visually selected based on negative phase reversals on bipolar montages. We analyzed the dipole position and moment of each spike using a single moving dipole and three-shell spherical head model. The dipoles were overlaid onto magnetic resonance (MR) images and divided into two groups based on two spike foci. RESULTS: The dipoles of the two groups were oriented either tangentially or radially to the scalp in close proximity to each other. The dipoles oriented radially were located underneath the electrode with a negative peak; those oriented tangentially were between electrodes with a negative and positive peak. The positions of tangential dipoles were more concentrated than those of radial dipoles. The epileptogenic regions corresponded to the dipole localizations. Surgical excisions were performed based on the results of electrocorticography. After surgery, two patients were seizure free, and one had rare seizures (follow-up period, 13-31 months). CONCLUSIONS: We showed that dipoles in close proximity but with different orientations projected two negative maxima on scalp EEG in three patients with extratemporal localization-related epilepsy. Equivalent current dipole analysis of individual interictal spikes can provide useful information about the epileptogenic zone in these patients. PMID- 11440344 TI - Frontal lobe seizure propagation: scalp and subdural EEG studies. AB - PURPOSE: To study propagation properties of focally originating frontal lobe seizures: principally direction and promptness of ictal spread. METHOD: Forty eight scalp EEG-recorded and 17 subdurally recorded seizures in two separate groups of patients were visually scrutinized. RESULTS: Initial propagation was directed more commonly to contiguous frontal cortex than to the opposite hemisphere in both studies: 58% and 86% for scalp and subdural recordings, respectively. Eighteen (38%) of scalp EEG seizures propagated within 5-10 s of apparent onset, whereas 16 (33%) did so after 11-20 s; no evidence of propagation could be discerned in the remaining 14 (29%). Of subdurally recorded attacks, only four (24%) propagated in the first 4 s; six (35%) did so after a delay of 5 10 s; six (35%) either failed to propagate or did so after > or =20-s latency. Latency to initial propagation was between 5 and 20 s in 41 (63%) of the 65 seizures in both studies. Ictal activity remained within the frontal lobe of origin for > or =10 s in 39 (81%) of scalp-recorded seizures and 11 (65%) of subdural seizures. It remained so confined for > or =15 s in 26 (54%) and nine (53%), respectively. Twenty-four (50%) of scalp-recorded seizures displayed evidence of opposite hemisphere (bisynchronous or other contralateral) involvement, occurring 5-10 s after onset in eight (17%) seizures and 10-20 s after onset in 16 (33%). Similarly, the subdural study documented spread to the opposite hemisphere in 11 (65%) of seizures with a latency ranging from 1 to 45 s. CONCLUSIONS: Frontal lobe seizures in this study propagated less promptly than reputed, and initial spread occurred more commonly to the frontal lobe of origin than to the opposite hemisphere. Such properties underlie the good lateralizing value of clinical semiology of frontal lobe seizures. PMID- 11440345 TI - Cerebrospinal fluid and serum neuron-specific enolase levels after febrile seizures. AB - PURPOSE: Neuron-specific enolase (NSE) has been established as a reliable marker of neuronal damage in various neurologic disorders. The aim of this study was to evaluate whether febrile seizures (FS) cause brain damage, based on the serum and cerebrospinal fluid (CSF) levels of NSE. METHODS: Fifty-three patients aged from 6 months to 7 years were enrolled. Among them, 36 patients had generalized seizures, and 17 had partial seizures. The maximal seizure duration was 90 min. Blood and CSF samples for measurement of NSE were obtained immediately after the seizure. NSE was measured using an enzyme immunoassay (EIA). RESULTS: Serum and CSF levels of NSE ranged up to 10 ng/mL, but very high levels were not observed. In patients with partial seizures, the NSE level in the CSF and the ratio of the CSF to serum NSE levels showed a strong correlation with seizure duration. Conversely, there were no correlations between NSE levels and seizure duration in the patients with generalized seizures. CONCLUSIONS: These results indicate that FS seldom cause severe neurologic damage, but prolonged partial seizures may cause slight neuronal injury. PMID- 11440346 TI - Temporal epileptogenesis: localizing value of scalp and subdural interictal and ictal EEG data. AB - PURPOSE: To determine the value of scalp epileptiform EEG data and subdural interictal spikes in localizing temporal epileptogenesis among patients requiring invasive recordings. For this delineation, we related such factors to site of subdural seizure origin in 27 consecutive patients. METHODS: Patients with temporal lobe epilepsy whose non-invasive lateralizing data were inconclusive and therefore required subdural electroencephalography were studied. All patients had (a) 24-h scalp telemetered EEGs, (b) adequate bitemporal subdural placements with an inferomesial line extending from a posterior burr hole anteriorly to <2.5 cm from anterior uncus and a lateral line reaching within 2.5 cm of the temporal tip, and (c) > or =2 subdurally recorded seizures. RESULTS: Three hundred one (96%) of 314 subdurally recorded clinical seizures involving all 27 patients arose from a discrete focus; 266 (85%) arose from mesial temporal regions, which was the origin of the majority of seizures in 24 (89%) patients. The majority of subdural seizures arose ipsilateral to the majority of scalp EEG spikes in 22 (81%) of 27, and most subdural seizures of 15 (75%) of 20 arose ipsilateral to scalp seizures. Lateralization of interictal subdural spikes correlated with that of subdural seizures in 74-92% of patients, depending on the method of spike compilation: for example, most subdural seizures arose from the same lobe of most consistent principal temporal spikes in 92% of patients. These indices of epileptogenesis also appeared more commonly on the side of effective (> or =90% improvement) temporal lobectomy than contralaterally in the following proportions: most consistent principal subdural spikes, 86% of patients ipsilateral vs. 9% contralateral; scalp-recorded clinical seizures, 55% vs. 18%; scalp EEG spikes, 45% vs. 9%. CONCLUSIONS: Even among patients whose scalp data are sufficiently complex to require invasive recording for clarification, lateralization of temporal scalp interictal and ictal epileptiform activity and subdural interictal spikes should be included when assessing the side of temporal epileptogenesis. PMID- 11440347 TI - Antiepileptogenesis and seizure prevention trials with antiepileptic drugs: meta analysis of controlled trials. AB - PURPOSE: To synthesize evidence concerning the effect of antiepileptic drugs (AEDs) for seizure prevention and to contrast their effectiveness for provoked versus unprovoked seizures. METHODS: Medline, Embase, and The Cochrane Clinical Trials Register were the primary sources of trials, but all trials found were included. Minimal requirements: seizure-prevention outcome given as fraction of cases; AED or control assigned by random or quasi-random mechanism. Single abstracter. Aggregate relative risk and heterogeneity evaluated using Mantel Haenszel analyses; random effects model used if heterogeneity was significant. RESULTS: Forty-seven trials evaluated seven drugs or combinations for preventing seizures associated with fever, alcohol, malaria, perinatal asphyxia, contrast media, tumors, craniotomy, and traumatic brain injury. Effective: Phenobarbital for recurrence of febrile seizures [relative risk (RR), 0.51; 95% confidence interval (CI), 0.32-0.82) and cerebral malaria (RR, 0.36; CI, 0.23-0.56). Diazepam for contrast media-associated seizures (RR, 0.10; CI, 0.01-0.79). Phenytoin for provoked seizures after craniotomy or traumatic brain injury (craniotomy: RR, 0.42; CI, 0.25-0.71; TBI: RR, 0.33; CI, 0.19-0.59). Carbamazepine for provoked seizures after traumatic brain injury (RR, 0.39; CI, 0.17-0.92). Lorazepam for alcohol-related seizures (RR, 0.12; CI, 0.04-0.40). More than 25% reduction ruled out valproate for unprovoked seizures after traumatic brain injury (RR, 1.28; CI, 0.76-2.16), and carbamazepine for unprovoked seizures after craniotomy (RR, 1.30; CI, 0.75-2.25). CONCLUSIONS: Effective or promising results predominate for provoked (acute, symptomatic) seizures. For unprovoked (epileptic) seizures, no drug has been shown to be effective, and some have had a clinically important effect ruled out. PMID- 11440348 TI - Visual function is stable in patients who continue long-term vigabatrin therapy: implications for clinical decision making. AB - PURPOSE: Vigabatrin (VGB) has been shown to cause visual field constriction and other forms of mild visual dysfunction. We determined the safety of continuing VGB therapy in patients who had received prolonged treatment (>2 years) with the drug by serially monitoring changes in visual function over a 1-year period of continued therapy. We also followed up patients who discontinued VGB to see whether alternative therapies are effective. METHODS: Fifteen of 17 patients who continued VGB therapy had visual-function testing (visual acuity, color vision, kinetic and static perimetry) every 3 months for 1 year. Eighteen patients who discontinued VGB were given alternative antiepileptic drugs (AEDs); their seizure responses were measured after > or =3 months of treatment. RESULTS: Patients continuing VGB showed no worsening of visual acuity, color vision, or visual field constriction beyond that measured in the initial test. Many patients who discontinued VGB had good seizure control with either newer or previously unsuccessful AEDs. CONCLUSIONS: For patients who have an excellent response to VGB and only mild visual changes, continued therapy may be safe with close visual monitoring. Patients who do not have a significant reduction in seizures or who experience considerable visual dysfunction with VGB may respond well to alternative therapies. PMID- 11440349 TI - Tiagabine: efficacy and safety in adjunctive treatment of partial seizures. AB - PURPOSE: To assess the efficacy and safety of tiagabine (TGB), a new antiepileptic drug (AED), as add-on therapy in patients with refractory partial seizures. METHODS: This response-dependent study used an open-label screening phase (in which patients were titrated to their optimal TGB dose, < or =64 mg/day) followed by a double-blind, placebo-controlled, crossover phase. Initial eligibility criteria included (a) seizures inadequately controlled by existing AEDs, and (b) six or more partial seizures during an 8-week baseline period. Patients showing benefit from TGB (> or =25% reduction in total seizure rate relative to baseline) were eligible for randomization into the double-blind phase, which comprised two 7-week assessment periods separated by a 3-week crossover period. RESULTS: Forty-four (50%) of the 88 enrolled patients entered the double-blind phase of the study during which there were significant reductions compared with placebo in all partial (p < 0.01), complex partial (p < 0.001), and secondarily generalized tonic-clonic seizure rates (p < 0.05). Thirty three percent of patients experienced a reduction of > or =50% in the all partial seizure rate. Eight (22%) patients receiving TGB during the double-blind phase reported adverse events, of which dizziness and incoordination were the most frequent. Three patients withdrew from treatment during the double-blind phase because of adverse events; two during treatment with TGB and one during treatment with placebo. TGB did not affect plasma concentrations of other coadministered AEDs. CONCLUSIONS: TGB was significantly better than placebo in terms of seizure rate reduction and was generally well-tolerated in patients with difficult to control seizures. PMID- 11440350 TI - Efficacy of clobazam as add-on therapy in patients with refractory partial epilepsy. AB - PURPOSE: Clobazam (CLB) has an important antiepileptic effect and is less expensive than the new antiepileptic drugs (AEDs), but still has not been considered as first-line drug in the treatment of epilepsy. We evaluated the efficacy of CLB as add-on therapy in patients with refractory partial epilepsy. METHODS: This was an open, retrospective study, conducted at the epilepsy clinic of our university hospital. All patients had chronic epilepsy and were being evaluated for epilepsy surgery. CLB was introduced as add-on therapy (starting with 10 mg/ day) in patients with previous failure of at least two AEDs. Information was obtained from clinical notes and follow-up visits. RESULTS: We evaluated 97 patients, 37 men and 60 women. Ages ranged from 15 to 70 years (mean, 35.8 years). Etiology of epilepsy was hippocampal atrophy in 67 (69%), cortical dysgenesis in nine (9.3%), and other etiologies in nine (9.3%). In 12 (12.3%) patients, the etiology of epilepsy was not identified despite clinical and neurologic investigation. Patients used CLB for a period ranging from 1 month to 7 years and 9 months (mean, 16.7 months) with doses ranging from 10 to 60 mg/day (mean, 29.7 mg/day). Seven (7.2%) patients were seizure free, 48 (49.4%) had > or =50% of improvement in seizure control, 39 (40.2%) had <50% of improvement in seizure control, and in three (3.1%), no data were available. CONCLUSIONS: We conclude that CLB may have efficacy equivalent to that of the new AEDs when used as add-on therapy in patients with refractory epilepsy. CLB should be considered an economic alternative in the treatment of patients with refractory epilepsy. PMID- 11440351 TI - Topiramate rapidly raises brain GABA in epilepsy patients. AB - PURPOSE: The short- and long-term pharmacodynamic effects of topiramate (TPM) on brain gammay-aminobutyric acid (GABA) metabolism were studied in patients with complex partial seizures. METHODS: In vivo measurements of GABA, homocarnosine, and pyrrolidinone were made of a 14-cc volume in the occipital cortex using 1H spectroscopy with a 2.1-Tesla magnetic resonance spectrometer and an 8-cm surface coil. Fifteen patients (four men) were studied serially after the first, oral dose (100 mg) of TPM. RESULTS: The first dose of TPM increased brain GABA within 1 h. Within 4 h, GABA was increased by 0.9 mM (95% CI, 0.7-1.1). Brain GABA remained elevated for > or =24 h. Pyrrolidinone and homocarnosine increased slowly during the first day. Daily TPM therapy (median, 300 mg; range, 200-500) increased GABA (0.3 mM; 95% CI, 0.1-0.5), homocarnosine (0.4 mM; 95% CI, 0.3 0.5), and pyrrolidinone (0.15 mM; 95% CI, 0.10-0.19), compared with levels before TPM. There was no dose response evident with daily TPM doses of 200-500 mg. CONCLUSIONS: TPM promptly elevates brain GABA and presumably offers partial protection against further seizures within hours of the first oral dose. Patients may expect to experience the effects of increased homocarnosine and pyrrolidinone within 24 h. PMID- 11440352 TI - Quality of life self-assessment as a function of lateralization of lesion in candidates for epilepsy surgery. AB - PURPOSE: This prospective study aimed to investigate the relationship(s) of the laterality of the epileptogenic lesion to personality factors, emotional processing, and the subjective experience of quality-of-life (QOL) self assessment in candidates for epilepsy surgery. METHODS: Patients who were candidates for epilepsy surgery were studied. Eighteen of them (aged 19-61 years) had localization-related epilepsy in the right temporal lobe (RTLE), 18 (aged 21 50 years) had localization-related epilepsy in the left temporal lobe (LTLE), and 20 were demographically matched normal subjects. The Spielberger Trait/State Anxiety questionnaire and the QOLIE-31 questionnaire for self-assessment of quality of life were used. One-way analysis of variance, Pearson correlations, and linear regression analyses were performed on group and anxiety levels and QOLIE variables. RESULTS: LTLE patients systematically showed higher levels of anxiety and lower self-estimates of the quality of their lives when compared with RTLE patients. All anxiety measures were highly correlated with Total QOL in LTLE (p < 0.05) but not in RTLE patients; however, different parameters of QOL showed different relationships with measures of anxiety. Results of multiple regression analyses suggested that the level of anxiety was relatively stable and less affected by QOL factors in LTLE as compared with RTLE patients (p = 0.03). CONCLUSIONS: A high level of anxiety shown by LTLE patients may represent a personality trait and cause a response bias in overreporting of negative symptoms and a decreased self-assessment of QOL. It may also present a presurgical risk factor, particularly for patients with LTLE lesions. Appropriate patient counseling is advised. PMID- 11440353 TI - High and low perceived self-control of epileptic seizures. AB - PURPOSE: To define behaviours and to identify psychological, demographic, and epilepsy-related variables associated with high as opposed to low perceived self control of seizures. METHOD: In a semistructured interview, 100 adults with intractable seizures were asked about their seizure precipitants and attempts at self-control of seizures. They also completed four psychological questionnaires. Latent Class Analysis was used to analyse the interview data to create two groups, High Controllers and Low Controllers, who were then compared on demographic, epilepsy, and psychological characteristics. RESULTS: Being able to identify and seeking out low-risk-for-seizure situations, avoiding high-risk-for seizure situations, and making attempts at seizure inhibition were seizure behaviours that discriminated High from Low Controllers. The general probability of being a High Controller was greater than that of being a Low Controller. Perceived high self-control of seizures was associated with low chance-health locus of control. For Low Controllers, current age, age at onset of seizures, and duration of epilepsy history were related to psychological variables. A significantly higher proportion of the Low Controllers than High Controllers were women. CONCLUSIONS: Many people with intractable seizures do not accept their epilepsy as a condition over which they have no control. Perceived self-control of seizures, however, involves a complex interaction between epilepsy and psychological factors, with health locus of control an apparently important discriminator between High and Low Controllers. PMID- 11440354 TI - Pharyngeal dysesthesias as an aura in temporal lobe epilepsy associated with amygdalar pathology. AB - PURPOSE: Pharyngeal dysesthetic auras are typically described with centrotemporal and opercular seizure-onset localizations. In this report we describe the fourth case in literature with temporal lobe seizures, apparently secondary to an amygdalar lesion on magnetic resonance imaging (MRI), presenting with prominent pharyngeal dysesthesias as the initial, or only, seizure manifestation. METHODS: Because of diagnostic uncertainty regarding the nature of the pharyngeal sensations, our case underwent prolonged extracranial video-EEG monitoring. RESULTS: Video-EEG information documented the epileptic origin of the dysesthesias and was concordant with the side and location of the amygdalar lesion. CONCLUSIONS: Pharyngeal dysesthetic auras may be produced by epileptic activity originating from the amygdala, and perhaps other mediotemporal structures. The underlying topography of this aura is not known with certainty, and it may reflect seizure spread from the amygdala and adjacent areas to the closely interconnected insular and opercular cortex, whose secondary activation could elicit similar sensations. PMID- 11440355 TI - Catamenial epilepsy and menopause. PMID- 11440356 TI - Minor head trauma unmasking asymptomatic lesions. PMID- 11440357 TI - Sleep and epilepsy: a role for nitric oxide. PMID- 11440358 TI - Diabetic vascular disease and the rising star of protein kinase C. PMID- 11440359 TI - Protein kinase C activation: isozyme-specific effects on metabolism and cardiovascular complications in diabetes. AB - Protein kinase C (PKC) is a family of multifunctional isoenzymes, activated by diacylglycerols (DAGs), which play a central role in signal transduction and intracellular crosstalk by phosphorylating at serine/threonine residues an array of substrates, including cell-surface receptors, enzymes, contractile proteins, transcription factors and other kinases. Individual isozymes vary in their pattern of tissue and subcellular distribution, function and Ca2+/phospholipid cofactor requirements, and in diabetes there is widespread activation of the DAG PKC pathway in metabolic, cardiovascular and renal tissues. In liver, muscle and adipose tissue, PKC isozymes have been implicated both as mediators and inhibitors of insulin action. Activation of DAG-sensitive PKC isoforms, such as PKC-theta and PKC-epsilon, down-regulates insulin receptor signalling and could be an important biochemical mechanism linking dysregulated lipid metabolism and insulin resistance in muscle. On the other hand, atypical PKC isozymes, such as PKC-zeta and PKC-lambda, have been identified as downstream targets of PI-3 kinase involved in insulin-stimulated glucose uptake, especially in adipocytes. Glucose-induced de novo synthesis of (palmitate-rich) DAG and sustained isozyme selective PKC activation (especially but not exclusively PKC-beta) has been strongly implicated in the pathogenesis of diabetic microangiopathy and macroangiopathy through a host of undesirable effects on endothelial function, VSM contractility and growth, angiogenesis, gene transcription (in part by MAP kinase activation) and vascular permeability. Interventions that increase DAG metabolism (e. g. vitamin E) and/or inhibit PKC isozymes (e. g. the beta selective inhibitor LY333531) ameliorate the biochemical and functional consequences of DAG-PKC activation in experimental diabetes, for example improving retinal blood flow and albuminuria in parallel with reductions in membrane-associated PKC isozyme activities. Thus, a greater understanding of the functional diversity and pathophysiological regulation of PKC isozymes is likely to have important clinical and therapeutic benefits. PMID- 11440360 TI - 15th Golgi lecture: from hyperglycaemia to the dysregulation of vascular remodelling in diabetes. AB - Hyperglycaemia has been shown to play a central part in diabetic vascular disease, which is also influenced by individual background. Hyperglycaemia initiates the pathogenetic sequence through a series of interrelated biochemical abnormalities, including increased flux through the polyol and hexosamine pathways, oxidative stress, AGE formation and protein kinase C activation. These abnormalities are capable of modifying the function of resident and non-resident vascular cells by changing their production pattern of several autocrine and paracrine factors, including growth, vasoactive and coagulation factors and adhesion molecules. These mediators profoundly impair the physiologic turnover of the vessel wall, thus leading to an abnormal process of vascular remodelling, with alterations in cell and matrix turnover and contacts, vascular tone and permeability and coagulation pattern. This process has distinct features depending on the target tissue. The hallmark of nephropathy is an abnormal accumulation of extracellular matrix within the mesangium, sustained by an upregulation of TGF-beta, possibly triggered by a local activation of the renin angiotensin system. The central pathological lesion in retinopathy is retinal ischaemia due to the formation of acellular capillaries. The resulting vascular endothelial growth factor-dependent neovascularization is a detrimental phenomenon leading to the formation of noncompetent vessels. Conversely, in macrovascular disease, arterial occlusion resulting from plaque formation with superimposed thrombosis elicits an angiogenic response which is impaired, but generates competent vessels, potentially compensating for reduced flow. Thus, upstream interventions interrupting the pathogenetic sequence at the level of hyperglycaemia (and related biochemical events) are the most effective, whereas downstream interventions should be targeted to the tissue affected. PMID- 11440361 TI - Physical training decreases plasma thrombomodulin in type I and type II diabetic patients. AB - AIMS/HYPOTHESIS: Endothelial damage is an early step in the pathogenesis of atherosclerosis and its improvement through physical training can contribute to the known reduction of cardiovascular risk associated with exercise. An increase in some endothelium-dependent haemostatic parameters, considered as markers of endothelial damage, has been observed in diabetic patients. METHODS: The effect of a three-month physical exercise programme on thrombomodulin, tissue factor pathway inhibitor, plasminogen activator inhibitor, tissue-type plasminogen activator and von-Willebrand factor was evaluated in 14 well-controlled patients with Type I (insulin-dependent) diabetes mellitus and 13 patients with Type II (non-insulin-dependent) diabetes mellitus (HbA1c 6.5 +/- 0.8 and 7.4 +/- 0.8%, respectively). A matched control group was also studied. RESULTS: Thrombomodulin at baseline was higher in both Type I and Type II diabetic patients than in their respective matched control subjects (50.0 +/- 16 vs 31.1 +/- 8.7 microg/l, p < 0.05; 51.0 +/- 10 vs 28.5 +/- 11 microg/l, p <0.05, respectively). After the exercise programme, thrombomodulin plasma concentrations had decreased (p < 0.05) in both groups of patients, with final thrombomodulin values being similar to those observed in their control groups (38.2 +/- 11 microg/l for Type I and 34.6 +/- 12 microg/l for Type II patients). The thrombomodulin decrement correlated with baseline thrombomodulin and VO2max increase in Type I diabetic patients. A decrease in tissue factor pathway inhibitor was also observed in Type II diabetic patients. CONCLUSION/INTERPRETATION: We conclude that the normalisation of plasma thrombomodulin concentrations in Type I and Type II diabetic patients after physical training might reflect the improvement in endothelial function associated with physical exercise. PMID- 11440362 TI - The influence of plasma glucose upon pulsatile ocular blood flow in subjects with type II diabetes mellitus. AB - AIMS/HYPOTHESIS: This study investigated the influence of plasma glucose upon pulsatile ocular blood flow in subjects with Type II (non-insulin-dependent) diabetes mellitus. METHODS: A total of 19 subjects with Type II diabetes and 8 normal control subjects undertook a meal tolerance test after an overnight fast. The pulsatile ocular blood flow, using the Ocular Blood Flow Tonometer, and plasma glucose concentrations were taken at times 0 min, 90 min and 240 min. Blood pressure and glycated haemoglobin concentrations, in the subjects with diabetes, were also measured at time 0 min. Pulsatile ocular blood flow and plasma glucose were also measured at times 0 and 90 min in 5 subjects with Type II diabetes mellitus who remained fasting. RESULTS: It was found that the subjects with diabetes who undertook the meal tolerance test showed a significant increase in both plasma glucose concentrations and pulsatile ocular blood flow from time 0-90 min, followed by a decrease from 90 min to the end of the session at 240 min. (p < 0.001 in each case). Regression analysis showed a significant correlation between the change in pulsatile ocular blood flow and the change in plasma glucose concentration (r = 0.671,p = 0.001). Control subjects showed no significant change in either plasma glucose or pulsatile ocular blood flow during the meal tolerance test. Subjects with diabetes mellitus who remained fasting also showed no significant change in pulsatile ocular blood flow or plasma glucose concentrations. No correlation was found between glycated haemoglobin concentrations or blood pressure and pulsatile ocular blood flow. CONCLUSION/INTERPRETATION: Pulsatile ocular blood flow is influenced by changes in plasma glucose concentrations in Type II diabetes mellitus, indicating that uncontrolled hyperglycaemia might result in a higher pulsatile ocular blood flow than might otherwise be expected. PMID- 11440363 TI - Oxidative stress could precede endothelial dysfunction and insulin resistance in Indian Mauritians with impaired glucose metabolism. AB - AIMS/HYPOTHESIS: To measure oxidative stress, endothelial dysfunction and insulin resistance in Indian Mauritians at different stages of development of Type II (non-insulin-dependent) diabetes mellitus. METHODS: Plasma total 8-epi-PGF2alpha, an indicator of oxidative stress, was determined in age-matched subjects with normal glucose metabolism (n = 39), impaired glucose tolerance (n = 14), newly diagnosed diabetes (n = 8) and established diabetes (n = 14). Plasma glucose and insulin were measured at baseline and 2 h following an oral glucose tolerance test. Endothelial function was assessed by non-invasive digital pulse wave photoplethysmography. RESULTS: Plasma 8-epi-PGF2alpha increased in subjects with impaired glucose tolerance (p < 0.05) compared with control subjects, and was even higher in newly diagnosed diabetic patients (p < 0.01) and established (p < 0.01) diabetic patients. A tendency towards reduced endothelial function in subjects with impaired glucose tolerance became significant in patients with newly diagnosed and established diabetes (p < 0.01), and was correlated with 8 epi-PGF2alpha (r = 0.36, p < 0.01). Insulin resistance (homeostasis model assessment) did not change in subjects with impaired glucose tolerance compared with control subjects, but increased in newly diagnosed (p < 0.01) and established (p < 0.001) diabetic subjects. The 8-epi-PGF2alpha was correlated with fasting glucose (r = 0.50, p < 0.001), triglycerides (r = 0.40, p < 0.001) and insulin resistance (r = 0.35, p < 0.001). CONCLUSION/INTERPRETATION: Oxidant stress is an early event in the evolution of Type II diabetes and could precede the development of endothelial dysfunction and insulin resistance. PMID- 11440364 TI - Transcription factor decoy for activator protein-1 (AP-1) inhibits high glucose- and angiotensin II-induced type 1 plasminogen activator inhibitor (PAI-1) gene expression in cultured human vascular smooth muscle cells. AB - AIMS/HYPOTHESIS: Multiple factors, including hyperglycaemia and angiotensin II (Ang II), stimulate plasminogen activator inhibitor-1 (PAI-1) gene expression in human vascular smooth muscle cells. This study tested the hypothesis that hyperglycaemia and Ang II stimulate PAI-1 gene expression through activator protein-1 (AP-1) binding sites. METHODS: We evaluated the role of AP-1 in PAI-1 gene expression in human vascular smooth muscle cells under high D-glucose and Ang II stimulation using a double-stranded cis-element AP-1 oligodeoxynucleotide (decoy ODN). RESULTS: Activator protein 1 activity was stimulated by high glucose and Ang II treatment and the AP-1 decoy ODN, but not a mismatched decoy ODN, competed for AP-1 activity. The increase in PAI-1 expression by high glucose and Ang II was significantly attenuated by the AP-1 decoy ODN (p <0.05 or p < 0.01). The increase in PAI-1 expression by high glucose and Ang II action on AP-1 sites was also confirmed by promoter analysis of PAI-1. Activator protein 1 activation in response to either high glucose or co-stimulation with high glucose and Ang II was inhibited completely by calphostin C (a PKC inhibitor) and partially by genistein (a protein tyrosine kinase inhibitor). CONCLUSION/INTERPRETATION: This study shows that high glucose and Ang II stimulate PAI-1 expression through AP-1 binding sites. Signal transduction after AP-1 activation by both high glucose and Ang II largely depends on PKC activation. These data indicate an important role for AP-1 in PAI-1 expression. PMID- 11440365 TI - Intra-uterine environment influences glomerular number and the acute renal adaptation to experimental diabetes. AB - AIMS/HYPOTHESIS: We sought to test the hypothesis of whether low birth weight rats would have reduced glomerular number, higher systolic blood pressure and an altered acute response to streptozotocin diabetes compared to normal birth weight rats. METHODS: Female offspring of Wistar rats fed an isocaloric diet containing either 6% casein (LPD) or 18% casein (NPD) in utero were studied. Birth weight, body weight, systolic blood pressure and urine albumin excretion were measured before and after streptozotocin diabetes. Glomerular number and volume were estimated after one week of diabetes. RESULTS: The LPD rats were of low birth weight (5.4 +/- 0.5 g vs 6.4 +/- 0.8 g, p < 0.0001) with higher systolic blood pressure (137 +/- 9mmHg vs 120 +/- 7 mmHg, p < 0.0001) and reduced glomerular number (17,435 +/- 2,074 vs 24,846 +/- 1,864, p < 0.0001). The LPD rats had smaller kidneys (0.925 +/- 0.009 g vs 1.200 +/- 0.173 g, p = 0.041) but similar glomerular volume to NPD control rats (1.11 +/- 0.15 x 10(6) microm3 vs 1.08 +/- 0.17 x 10(6) microm3). After 1 week of diabetes LPD rats had a greater proportional increase in renal size (diabetes 50 +/- 12 % vs control 20 +/- 4%, p = 0.003). Insulin suppressed renal hypertrophy in both LPD and NPD rats but failed to suppress glomerular hypertrophy in LPD rats (1.48 +/- 0.21 x 10(6) microm3 vs 1.03 +/- 0.23 x 10(6) microm3 p = 0.015). CONCLUSION/INTERPRETATION: Abnormal intra-uterine environment reduces both renal size and glomerular number and influences the acute renal adaptation to experimental diabetes. PMID- 11440366 TI - Type I diabetes mellitus does not alter initial splanchnic glucose extraction or hepatic UDP-glucose flux during enteral glucose administration. AB - AIMS/HYPOTHESIS: Our aim was to determine whether an alteration in splanchnic glucose metabolism could contribute to postprandial hyperglycaemia in people with Type I (insulin-dependent) diabetes mellitus. METHODS: Splanchnic glucose extraction, hepatic glycogen synthesis and endogenous glucose production were compared in 8 Type I diabetic patients and in 11 control subjects. Endogenous hormone secretion was inhibited with somatostatin while insulin (approximately 550 pmol/l) and glucagon (approximately 130 ng/l) concentrations were matched with exogenous hormone infusions. Glucose containing [3-3H] glucose was infused into the duodenum at a rate of 20 micromol.kg(-1).min(-1). Plasma glucose concentrations were maintained at about 8.5 mmol/l in both groups by means of a separate variable intravenous glucose infusion. RESULTS: Initial splanchnic glucose uptake, calculated by subtracting the systemic rate of appearance of [3 3H] glucose from the rate of infusion of [3-3H] glucose into the duodenum, did not differ in the diabetic and non-diabetic patients (4.1 +/- 0.8 vs 3.0 +/- 1.0 micromol/kg/min). In addition, hepatic glycogen synthesis, measured using the acetaminophen glucuronide method did not differ (10.7 +/- 2.4 vs 10.1 +/- 2.7 micromol.kg(-1).min(-1)). On the other hand, suppression of endogenous glucose production, measured by an intravenous infusion of [6,6-2H2] glucose, was greater (p < 0.05) in the diabetic than in the non-diabetic subjects (1.7 +/- 1.6 vs 5.8 +/- 1.9 micromol.kg(-1).min(-1)). CONCLUSION/INTERPRETATION: When glucose, insulin and glucagon concentrations are matched in individuals with relatively good chronic glycaemic control, Type I diabetes does not alter initial splanchnic glucose uptake of enterally delivered glucose or hepatic glycogen synthesis. Alterations in splanchnic glucose metabolism are not likely to contribute to postprandial hyperglycaemia in people with well controlled Type I diabetes. PMID- 11440367 TI - Differential mechanisms of glucose and palmitate in augmentation of insulin secretion in mouse pancreatic islets. AB - AIMS/HYPOTHESIS: To assess the possible importance of saturated fatty acids in glucose amplification of K+ATP channel-independent insulin secretion. METHODS: Insulin release from perifused pancreatic islets of NMRI mice was determined by radioimmunoassay. RESULTS: In the presence of K+ (20 mmol/l) and diazoxide (250 micromol/l), which stimulates Ca2+ influx and opens K+ATP channels, palmitate (165 micromol/l total; 1.2 micromol/l free) increased insulin secretion at 3.3, 10 and 16.7 mmol/l glucose while glucose (10; 16.7 mmol/l) did not increase insulin secretion. In the presence of K+ (60 mmol/l) and diazoxide (250 micromol/l), glucose (10; 16.7 mmol/l) stimulation of K+ATP channel-independent insulin secretion increased, whereas the effectiveness of palmitate (165 micromol/l total; 1.2 micromol/l free) on insulin secretion at both 3.3, 10 or 16.7 mmol/l glucose was reduced. Palmitate thereby mimicked the stimulatory pattern of the protein kinase C activator, 12-O-tetradecanoylphorbol 13-acetate (0.16 micromol/l), which also failed to increase insulin secretion at maximum depolarising concentrations of K+ (60 mmol/l). Furthermore, the protein kinase C inhibitor calphostin C (1 micromol/1), led to a complete suppression of the effects of both palmitate (165 micromol/l total; 1.2 micromol/l free) and myristate (165 micromol/l total; 2.4 micromol/l free) stimulation of glucose (16.7 mmol/l)-induced insulin secretion. Calphostin C (1 micromol/l), however, failed to affect insulin secretion induced by glucose (16.7 mmol/l). CONCLUSION/INTERPRETATION: These data suggest that glucose could increase insulin secretion independently of saturated fatty acids like palmitate and myristate, which amplify glucose-induced insulin secretion by activation of protein kinase C. PMID- 11440368 TI - Effect of repaglinide on cloned beta cell, cardiac and smooth muscle types of ATP sensitive potassium channels. AB - AIMS/HYPOTHESIS: The carbamoylbenzoic acid derivative repaglinide is a potent short-acting insulin secretagogue that acts by closing ATP-sensitive potassium (KATP) channels in the plasma membrane of the pancreatic beta cell. In this paper we investigated. the specificity of repaglinide for three types of cloned (KATP) channel composed of the inwardly rectifying potassium channel Kir6.2 and either the sulphonylurea receptor SUR1, SUR2A or SUR2B, corresponding to the beta cell, cardiac and either smooth muscle types of KATP channel, respectively. METHODS: The action of the drug was studied by whole-cell current recordings of KATP channels expressed either in Xenopus oocytes or mammalian cells (HEK293). We also used inside-out macropatches excised from Xenopus oocytes for detailed analysis of repaglinide action. RESULTS: The drug blocked all three types of KATP channel with similar potency, by interacting with a low-affinity site on the pore-forming subunit of the channel (Kir6.2: half-maximal inhibition 230 micromol/l) and with a high-affinity site on the regulatory subunit, the sulphonylurea receptor (SUR: half-maximal inhibition 2-8 nmol/l). There was no difference in potency between channels containing SUR1, SUR2A or SUR2B. MgADP potentiated the inhibitory effect of repaglinide on Kir6.2/SUR1 and (to a lesser extent) Kir6.2/SUR2B, but not on Kir6.2/SUR2A. CONCLUSION/INTERPRETATION: Repaglinide interacts with a site common to all three types of sulphonylurea receptor leading to inhibition of the KATP channel. The fact that MgADP potentiated this effect in the case of the beta cell, but not cardiac, type of channel could help explain why the drug shows no adverse cardiovascular side-effects in vivo. PMID- 11440369 TI - Intravitreal injection of plasminogen kringle 5, an endogenous angiogenic inhibitor, arrests retinal neovascularization in rats. AB - AIMS/HYPOTHESIS: Plasminogen kringle 5 is an endogenous angiogenic inhibitor. The purpose of the present study was to explore the potential application of kringle 5 in the treatment of retinal neovascularization. METHODS: Plasminogen kringle 5 was expressed in E. coli and affinity-purified. Its anti-angiogenic activity was determined in cultured primary human capillary endothelial cells. Retinal neovascularization was induced in newborn rats by exposure to hyperoxia and then normoxia. Kringle 5 was intravitreally injected into the rat model. Retinal neovascularization was visualized by fluorescein angiography on flat-mounted retina and quantified by counting preretinal vascular cells. RESULTS: Plasminogen kringle 5 inhibited primary endothelial cells but not retinal neuronal cells, suggesting cell type-specific inhibition. The oxygen-induced retinopathy rat model showed an over-expression of vascular endothelial growth factor, preretinal neovascularization and haemorrhage. Intravitreal injection of kringle 5 before the development of neovascularization resulted in fewer neovascular tufts and pre retinal vascular cells than in control rats with PBS injection (p < 0.01). Moreover, injection of kringle 5 after the development of neovascularization inhibited the increase in the preretinal vascular cells (p < 0.05). These results suggest that kringle 5 both prevents the development and arrests the progression of retinal neovascularization. The injection of kringle 5 did not result in any detectable inflammatory response in the retina or histological toxicity to retina neurons and pre-existing vessels. CONCLUSION/INTERPRETATION: These observations suggest that intravitreal delivery of angiogenic inhibitors could have therapeutic benefits in neovascular diseases of the retina. PMID- 11440370 TI - Increased rate of lipid peroxidation and protein carbonylation in experimental diabetic pregnancy. AB - AIMS/HYPOTHESIS: Maternal Type I (insulin-dependent) diabetes mellitus is associated with an increased risk for fetal malformations and spontaneous abortions. Although the pathogenic mechanism is not fully understood, reactive oxygen species have been shown to contribute to the pathogenesis in experimental studies. By measuring 8-iso-PGF2alpha and protein carbonyls, radical oxygen damage to lipids and proteins can be estimated. The aim of this study was to investigate the status of lipid peroxidation and protein carbonylation in mothers and fetuses in experimental diabetic pregnancy. METHODS: Non-pregnant and pregnant rats with and without streptozotocin-induced diabetes were studied after 4 weeks of diabetes or at gestational day 19, respectively. Gross morphology of the offspring was studied and 24 h urine, plasma, amniotic fluid, maternal and fetal livers were collected. Concentrations of 8-iso-PGF2alpha, 15-keto-DH PGF2alpha and other oxidative stress variables were measured. RESULTS: Malformation and resorption rates were increased in diabetic litters, whereas fetal weights were decreased in the control rats. There were no statistically significant differences in maternal plasma concentrations of 8-iso-PGF2alpha, but plasma protein carbonyl content was increased in the diabetic groups. Pregnancy increased 24 h urinary excretion of 8-iso-PGF2alpha in diabetic rats but not in the control rats. There was no difference in the amniotic fluid concentration of 8-iso-PGF2alpha between the normal and the diabetic group. However, in the diabetic group there was a correlation between the uterine horn concentration of 8-iso-PGF2alpha and the percentage of resorptions. CONCLUSIONS/INTERPRETATION: In diabetic pregnancy, both diabetes and pregnancy are promoting oxygen radical damage. Fetal oxidative stress markers do not clearly reflect fetal morphology. PMID- 11440371 TI - Routine mutation screening of HNF-1alpha and GCK genes in MODY diagnosis: how effective are the techniques of DHPLC and direct sequencing used in combination? AB - AIMS/HYPOTHESIS: Mutations in the hepatocyte nuclear factor (HNF)-1alpha and glucokinase (GCK) genes are the major causes of monogenic forms of Type II (non insulin-dependent) diabetes mellitus (Maturity-Onset Diabetes of the Young subtypes, MODY). We evaluated the effectiveness of fluorescent single-strand conformation polymorphism (F-SSCP), denaturing high-performance liquid chromatography (DHPLC) and sequencing based mutation detection in the molecular diagnosis of MODY. Our goal is to identify a rapid, efficient and cost effective mutation detection method for the molecular diagnosis of MODY and other human genetic disorders. METHODS: We evaluated the accuracy of DHPLC in screening for MODY 2 and 3 mutations. In addition, we compared the sensitivity, specificity, cost, handling time and analysis time of fluorescent single-strand conformation polymorphism, denaturing high-performance liquid chromatography and direct sequencing screening methods. RESULTS: Denaturing high-performance liquid chromatography is a recently developed method for mutation detection. It is cost effective, powerful and reliable and quite suitable for 22 out of the 24 fragments required for MODY 2 and 3 testing. However, exons 1 and 7 of the HNF 1alpha gene are very polymorphic and so direct sequencing is faster as well as more efficient and reliable. CONCLUSION/INTERPRETATION: Our results suggest that combining denaturing high-performance liquid chromatography and direct sequencing is a good approach for the routine detection of HNF-1alpha and GCK mutations in MODY families. Denaturing high-performance liquid chromatography appears to be a powerful tool in genetic testing and the method could be applied to the molecular diagnosis of other human genetic diseases. PMID- 11440372 TI - Analysis of the lamin A/C gene as a candidate for type II diabetes susceptibility in Pima Indians. AB - AIMS/HYPOTHESIS: Lamin A/C (LMNA) is located within a region on chromosome 1q that has been linked with Type II (non-insulin-dependent) diabetes mellitus in Pima Indians. Rare mutations in exon 8 of LMNA underlie Dunnigan-Type familial partial lipodystrophy, a disease characterized by regional adipocyte degeneration and frequently accompanied by insulin resistance, glucose intolerance, and diabetes. A more common variant in exon 10 (3408C/T) has recently been associated with obesity in non-diabetic aboriginal Canadian subjects. Because obesity is a strongly predisposing factor for Type II diabetes, we hypothesized that the LMNA 3408C/T variant could be associated with diabetes and body mass index in Pima Indians. METHODS: To determine whether the LMNA 3408C/T variant contributes to Type II diabetes susceptibility, we genotyped the polymorphism in 1,338 Pimas using allelic discrimination technology. The locus was screened for additional variants in 20 diabetic Pima Indians and non-diabetic Pima Indians using denaturing high performance liquid chromatography and dideoxy sequencing. RESULTS: We found no evidence for association of 3408C/T with diabetes, body mass index, total cholesterol, HDL cholesterol, triglycerides, leptin concentrations, or indices of insulin sensitivity and secretion. Subsequent screening of the remaining LMNA exons and flanking sequences revealed only rare variants in intron 4 and the 3'UTR, showing no frequency differences between diabetic and non diabetic Pima Indians. We reassessed the linkage with diabetes following adjustment for the LMNA 3408C/T variant; adjustment for the effects of LMNA did not substantially modify the evidence for linkage. CONCLUSION/INTERPRETATION: We conclude that the LMNA 3408C/T variant probably does not play a role in susceptibility to diabetes or obesity in Pima Indians. PMID- 11440373 TI - --to: Albareda M et al. (2000) assessment of insulin sensitivity and beta-cell function from measurements in the fasting state and during an oral glucose tolerance test. Diabetologia 43: 1507-1511. PMID- 11440374 TI - --to: Bergman RN (2000) non-esterified fatty acids and the liver: why is insulin secreted into the portal vein? Diabetologia 43: 946-952. PMID- 11440375 TI - Munchausen's syndrome manifesting as factitious hypoglycaemia. PMID- 11440376 TI - No evidence for diabetes-associated mutations of PEK/EIF2AK3 gene in French patients with early-onset type II diabetes. PMID- 11440377 TI - The beta-cell Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) beta3 isoform containing a proline-rich tandem repeat in the association domain can be found in the human genome. PMID- 11440378 TI - The elevated proinsulin-to-insulin ratio is associated with oxidative stress in Down's syndrome. PMID- 11440379 TI - Report about a three months' stay in Milan supported by an EFSD MSD Travel Fellowship for Young Scientists 1999. PMID- 11440380 TI - Chlamydia trachomatis in andrologic patients--direct and indirect detection. AB - BACKGROUND: Chlamydia trachomatis is considered to be the most common sexually transmitted disease in Germany. It is currently unclear whether chlamydial infection causes pathological conditions of the male accessory glands with consequences for male infertility. PATIENTS AND METHODS: Within the framework of several prospective studies the association between sperm quality, male accessory gland function and infection with C. trachomatis was investigated in men of couples with unexplained infertility. Chlamydial infection was determined by serologic methods and by proof of Chlamydia-specific DNA. As a marker of infection the direct determination of granulocytes in the ejaculate or the measurement of the polymorphonuclear (PMN) elastase concentration was used. The male accessory gland function was evaluated using the markers fructose, citric acid and alpha-glucosidase in the seminal plasma. RESULTS: Chlamydia-specific DNA in the ejaculate was present in between 3-5% of the subjects, which corresponds to its prevalence in the normal population. Chlamydia IgA antibodies were demonstrated with a frequency of 38% in seminal plasma (n = 834) using a genus specific test (rELISA). Using other species-specific tests (MIF, SeroCT, IgA pELISA and ImmunoComb), Chlamydia IgA antibodies were found at frequencies of between 8 and 22%. CONCLUSION: Only in a few individual cases was it possible to show a connection between reduced sperm quality, disturbed male accessory gland function and indication of infection with Chlamydia, bacteria or Ureaplasma. PMID- 11440381 TI - Alveolar liver echinococcosis: a comparative study of three imaging techniques. AB - BACKGROUND: We compared the imaging findings in patients with alveolar liver echinococcosis using ultrasound (US), computerized tomography (CT) and magnetic resonance imaging (MRI) in a prospective study. PATIENTS AND METHODS: 30 patients with alveolar echinococcosis (AE) were examined with the above imaging techniques. RESULTS: 30 lesions were detected with all three methods and most lesions (n = 55) were detected with CT. Calcifications were seen in 15 lesions with US, in 21 with CT and in 16 with MRI. MRI best detected necrotic areas and multivesicuLar structures. CONCLUSION: US is the screening method of choice and should primarily be complemented by CT due to its ability to detect the greatest number of lesions and clear demarcation of the characteristic calcifications. MRI may facilitate the diagnosis in uncertain cases with noncalcified or partially calcified lesions by showing the characteristic multivesicular structure, necrotic areas and proximity to vascular structures. PMID- 11440382 TI - Antimicrobial expenditures and usage at four university hospitals. Baden Wurttemberg Interuniversity Study Group. AB - BACKGROUND: The increasing use of antimicrobiaL drugs is resulting in enormous hospital expenditures. Careful assessment of inappropriate prescribing and a search for more cost-effective treatment strategies are urgently required. Comparisons between hospitals should help identify areas of inappropriate prescribing as well as effective drug use programs, but such analyses may be severely biased if the impact of different case-mixes is not recognized. PATIENTS AND METHODS: We studied antimicrobial usage and expenditures at four state university hospitals in southwestern Germany and assessed the significance of differences between hospital services after adjustment for patient variables. A prevalence survey was done with review of 2,254 charts of patients admitted to the surgical, medical and pediatric services to obtain information on antimicrobial drug prescription and expenditures in the week preceding the survey. RESULTS: According to pharmacy data for the year 1994, maximal differences between these hospitals in the antibiotic costs per patient-day were 1.9-fold (surgical services), 1.5-fold (medical services), and 1.6-fold (pediatric services). In a multivariate analysis, adjusted antibiotic prescription prevalence rates did not differ for medical and pediatric service patients, but did differ for surgical service patients (p = 0.03). Similarly, adjusted expenditures per patient-week differed significantly between hospitals for surgical service patients (p = 0.001), but only marginally for medical (p = 0.14) and pediatric (p = 0.05) service patients. The adjusted difference in expenditures between surgical departments was as large as 2.8-fold (95% CI 1.8 to 4.3) and was primarily related to preferential use of expensive iv antimicrobial drugs. CONCLUSION: In two hospitals, lowest expenditures in either surgery or medicine were associated with active antimicrobial drug use programs suggesting an impact of these programs on drug use and expenditures limited to these services. The identification of such large patient-mix unrelated differences in antimicrobial usage and expenditures offers opportunities for quality improvements and cost reduction. PMID- 11440383 TI - Enterovirus infections in Germany: comparative evaluation of different laboratory diagnostic methods. AB - BACKGROUND: The diagnosis of an enterovirus infection may be achieved through direct virus detection from fecal or cerebrospinal fluid (CSF) samples by virus isolation or PCR. Serologically, a significant rise in antibody titer may be detected and different enteroviral types can be differentiated using the neutralization assay. PATIENTS AND METHODS: We investigated the contribution of these different laboratory parameters to the diagnosis of enterovirus infections occurring in the Frankfurt am Main area during the years 1997 to 1999, including an echovirus 30 outbreak in a group of children with aseptic meningitis in 1997. Samples were referred from 1,013 patients; virus isolation was attempted from 579 CSF specimens and from 400 stool samples. 208 CSF samples were tested by PCR. RESULTS: During the echovirus 30 outbreak we identified 22.3% of samples as positive, almost exclusively echovirus 30. In 1998 only 7.1% of samples were positive and a rather broad range of agents was isolated. In 1999 10.4% were positive, predominantly coxsackie B5 and echovirus 11. We could show that in acute enterovirus infections, virus detection by cell cuLture and PCR is superior to serological methods (neutralization assay and IgM assay). For virus isolation, there was a higher rate of positives from stool compared to CSF (1997: 27.8% versus 25%; 1998: 14.4% versus 3%; 1999: 17.9% versus 8.5%). When comparing PCR and virus isolation from the CSF, the former yielded a higher rate of positive results but was not clearly superior to virus isolation from CSF. CONCLUSION: The recommended method for the diagnosis of acute enterovirus infections is virus isolation from feces. In cases of suspected aseptic meningitis virus isolation and PCR are valuable for the direct detection of virus in CSF. PMID- 11440384 TI - Nosocomial Kikuchi's disease--a search for herpesvirus sequences in lymph node tissues using PCR. AB - BACKGROUND: Histiocytic necrotizing lymphadenitis, also known as Kikuchi's disease (KD), is a rare disease. Fever and lymphadenopathies with characteristic pathologic features are present. The etiology of this disease remains undetermined. Since the disorder is self-limiting, different viruses have been implicated as the causative agent. PATIENTS AND METHODS: Seven cases of KD were studied. Three patients acquired the disease nosocomially, three had community acquired KD and one case was associated with systemic lupus erythematosus. PCR was performed on DNA extracted from lymph node tissues in order to detect herpesvirus-specific DNA sequences: herpes simpLex virus type 1 and 2 (HSV1-2), varicella zoster virus (VZV), human cytomegalovirus (HCMV), human herpesvirus 6 (HHV6), Epstein-Barr virus (EBV) and human herpesvirus 8 (HHV8). RESULTS: Viral DNA was not detected in any of the lymph node tissues from the seven cases of KD. CONCLUSION: We conclude that these herpesviruses were not involved in the etiology of the three cases of nosocomial KD, nor in the other four cases of KD investigated in this study. PMID- 11440385 TI - Magnetic resonance imaging in diagnosis of childhood central nervous system tuberculosis. AB - BACKGROUND: We investigated the role of magnetic resonance imaging (MRI) in diagnosis of central nervous system (CNS) tuberculosis. PATIENTS AND METHODS: The MRI findings (T1, T2 and T1 sequence after contrast) of 11 children with CNS tuberculosis were evaluated and the diagnostic value of contrast enhanced MRI and its correlation with the clinical findings were investigated. RESULTS: The median age of the patients was 6 years; seven were male and four female. MRI findings of the cases were meningeal enhancement (90.9%), hydrocephalus (63.6%), infarction (45.5%), tuberculomas (27.2%), cranial nerve involvement (27.2%) and severe cortical atrophy (9.1%). MRI provided early diagnosis in two cases by demonstrating tuberculomas, and supported the diagnosis in seven cases. However, in two cases MRI findings were inadequate to diagnose CNS tuberculosis, especially at the beginning. CONCLUSION: Contrast enhanced MRI is a very useful technique for diagnosing CNS tuberculosis, particularly by demonstrating the localized lesions, meningeal enhancement and the brain stem. However, a family history of tuberculosis, chest X-ray findings and the clinical picture are still very important for diagnosis. PMID- 11440386 TI - Clinical relevance of CagA-specific antibodies related to CagA status of Helicobacter pylori isolates using immunofluorescence test and PCR. AB - BACKGROUND: The cagA (cytotoxin-associated gene A) protein is found in about 50% of Helicobacter pylori strains; its clinical relevance in gastroduodenal disease is uncertain. PATIENTS AND METHODS: The frequency of IgG antibodies to cagA was studied by using a commercial Western blot assay in sera of 189 patients with endoscopically and histologically confirmed gastroduodenal disease. In addition, 38 H. pylori strains isolated from biopsies were analyzed by immunofluorescence test (IFT) and PCR for detection of cagA protein and cagA gene sequences, respectively. RESULTS: 54.3-60.0% of all patients with gastrointestinal diseases (chronic gastritis, gastric or duodenal ulcer and chronic duodenitis) and 28.6% with a normal mucosa were found to be positive for anti-cagA IgG antibodies. There was no significant difference in anti-cagA IgG seroprevalence between the different clinical entities. CagA-positive (cagA+) H. pylori strains were detected in 44.7% and 50% of the 38 isolates by PCR and IFT, respectively. 22 of 23 patients infected with cagA+ strains had anti-cagA antibodies. Using PCR as a gold standard, the sensitivity and specificity of the cagA IgG Western blot were 100.0% and 35.0%, respectively; the sensitivity and specificity of the cagA IFT were 76.5% and 71.4%, respectively. The incidence of the cagA+ H. pylori strains detected either by PCR or IFT was significantly higher (p < 0.05 and p < 0.01, respectiveLy) in patients with chronic duodenitis, gastric or duodenal ulcer compared to patients with chronic gastritis (66.7%, 80% and 30.4%, respectiveLy). CONCLUSION: In this study the cagA-specific serological status in H. pylori infections as diagnosed by IgG Western blot was of no predictive value for severity of disease. In contrast, the cagA status of H. pylori isolates, diagnosed by IFT or PCR, was a predictive marker for severe disease and, therefore, also of clinical relevance in the assessment of the virulence of the infecting strain. PMID- 11440387 TI - Neutralization activity and persistence of antibodies induced in response to vaccination with a novel mumps strain, RIT 4385. AB - BACKGROUND: We have previously shown that a new measles, mumps, rubella (MMR) vaccine, Priorix, with a novel mumps component elicits anti-mumps antibody titers comparable to the licensed M-M-R II vaccine. MATERIALS AND METHODS: To ensure that these antibodies had neutralizing activity against wild-type mumps virus, sera were prepared 2 and 18 months after vaccination of 12-24-month-old infants with either Priorix or M-M-R II and ELISA antibody titers and neutralizing activity were determined. RESULTS: After 2 months, Priorix and M-M-R II vaccines elicited comparable ELISA antibody titers and neutralizing activity. At 18 months ELISA seropositivity rates were 80-81% and 94-96% of vaccinees had neutralizing activity. All ELISA seropositives had neutralizing activity. CONCLUSION: These data suggest that both vaccines provided equivalent protection against mumps over this 18-month period. PMID- 11440388 TI - Typing of Streptococcus pyogenes strains isolated from throat infections in the region of Aachen, Germany. AB - BACKGROUND: Changes in the epidemiology of Streptococcus pyogenes infections may be associated with the introduction and reappearance of individual serotypes within a population. MATERIALS AND METHODS: Typing of 216 consecutive isolates of S. pyogenes from patients with pharyngitis in the region of Aachen, Germany, was performed by sequencing the emm gene, slide-agglutination of the T-antigen and determining the serum opacity reaction (SOR). RESULTS: All 216 isolates were unequivocally emm-typable. emm1 was most common (18.5%), foLlowed by emm12 (15.7%), emm3 (14.4%) and emm28 (13.9%). Only four isolates contained newly validated emm types: emm89 or emm94 were harbored by two isolates each. In one isolate, the sequence type s104 was found. CONCLUSION: Despite an anticipated selective pressure, the prevalence of emm1 among isolates from throat infections in northwestern Germany remains high, but does not reflect the predominance of emm1 among invasive isolates in Germany. PMID- 11440389 TI - Clinical expression of 'silent' hepatitis B virus infection in a patient with visceral leishmaniasis. AB - A 69-year-old male was hospitalized in January 1999 because of visceral leishmaniasis. He had also suffered from anti-hepatitis C virus (HCV)-positive chronic hepatitis for years. All serum hepatitis B virus (HBV) antigens and antibodies were negative except for anti-HBc. The patient was treated with amphotericin B cholesteryl sulfate (2 mg/kg twice a day for 7 days, iv). Fever disappeared on the 3rd day of treatment, the clinical condition improved rapidly and the patient recovered. In May 1999 the patient developed icteric HBsAg negative acute hepatitis (aspartate aminotransferase 722 U/l; alanine aminotransferase 988 U/l). Anti-HBc IgM was positive and HBV-DNA was detected in serum by PCR. Anti-HAV IgM was negative. A serum sample obtained on presentation and stored at -80 degrees C was retrospectively tested and found positive for HBV DNA. In July 1999, complete remission of acute hepatitis and seroconversion to anti-HBs was observed. We suppose that a moderate depression of the immune system, probably associated with leishmaniasis, may have enhanced HBV replication in the patient who had an HBsAg-negative 'silent' HBV infection. Restoration of the immune system after successful antiprotozoan therapy might have induced cell mediated necrosis of the HBV-infected hepatocytes and seroconversion to anti-HBs. PMID- 11440390 TI - Cervical lymphadenitis--a rare case of focal listeriosis. AB - Cervical lymphadenopathy is a condition with a broad differential diagnosis. Malignant tumors, allergic reactions, autoimmune diseases as well as various infectious agents can be the cause. We report the case of a 75-year-old man who had a cervical lymphadenopathy without symptoms of infection. The draining lymph nodes were excised. Histology revealed a granulomatous abscess and culture yielded pure growth of Listeria monocytogenes. Because food is occasionally contaminated with L. monocytogenes, the pathogenesis of this condition may be similar to that of lymph node tuberculosis. PMID- 11440391 TI - Transient fungemia due to Rhodotorula rubra in a cancer patient: case report and review of the literature. AB - A unique case of Rhodotorula rubra transient fungemia in a post-chemotherapy, febrile neutropenic patient with colon cancer, suffering from gastrointestinal mucositis, is described. The fungus was isolated repeatedly from his blood. However, all signs and symptoms of the infection disappeared, without antifungal treatment, as soon as neutropenia and mucositis, both of short duration, resolved. Restoration of the patient's defense mechanisms was adequate for disappearance of the fungus from the patient's blood and full recovery. PMID- 11440392 TI - Prosthetic valve endocarditis due to Candida tropicalis complicated by multiple pseudoaneurysms. AB - Candida endocarditis is an unusual but severe complication caused by Candida albicans or other fungal species. We describe a case of prosthetic valve endocarditis due to Candida tropicalis, complicated by multiple pseudoaneurysms. PMID- 11440393 TI - Crystalloids: an iatrogenic source of multiple organ failure? PMID- 11440394 TI - Antibody response to influenza vaccine in children with bronchopulmonary dysplasia. PMID- 11440395 TI - What was your diagnosis? Congenital tricuspid dysplasia with concurrent Ebstein's anomaly. PMID- 11440396 TI - Lumbosacral osteochondrosis: radiological features and surgical management in 34 dogs. AB - Lumbosacral osteochondrosis has been diagnosed with increasing frequency over the past few years. Nevertheless, poor recognition of the condition continues to lead to frequent misdiagnosis. A study was therefore undertaken over a 12-year period (1986 to 1998) to describe the use of a variety of radiological studies to define the syndrome and the use of different surgical techniques for optimal treatment of the condition. Dogs with clinical signs of cauda equina neuropathy and radiological signs of lumbosacral osteochondrosis were older than 14 months (mean age 6.3 years). German shepherd dogs, boxers and rottweilers were heavily represented. Of the 34 dogs with osteochondrosis in this study, the lesion appeared to be related to the sacrum in 31 dogs (91 per cent) and the seventh lumbar vertebra in the remaining three dogs (9 per cent). Twenty-seven male and seven female dogs were affected (4:1). Out of the 34 dogs, six (18 per cent) were treated conservatively and the remaining 28 (82 per cent) were treated surgically. The outcome of surgical treatment was considered to be satisfactory in 24 (86 per cent) of these latter dogs. PMID- 11440397 TI - 'Battered pets': non-accidental physical injuries found in dogs and cats. AB - Records of 243 cases of non-accidental injury (NAI) in dogs, and 182 cases in cats, submitted by a sample of small animal practitioners in the UK, revealed a wide range of injuries. These included bruises, fractures, repetitive injuries, burns and scalds, stab and incised wounds, poisoning, asphyxiation and drowning (which showed remarkable similarities to NAI in children), as well as sexual abuse and injuries specifically caused by firearms. Traumatic skeletal injuries in the dogs were more commonly found in the anterior part of the skeleton, in comparison with those resulting from road traffic accidents. Young male dogs and young cats were particularly at risk of NAI. A moderately increased risk was identified in the Staffordshire bull terrier, cross-breed dogs and the domestic shorthaired cat, whereas the Labrador retriever showed a decreased risk. No single injury or group of injuries, when divorced from the circumstances surrounding a suspect case, could be considered to indicate, conclusively, NAI. Repetitive injuries, however, were highly suggestive of NAI. PMID- 11440398 TI - Malassezia and Candida infections in bull terriers with lethal acrodermatitis. AB - In 12 cases of lethal acrodermatitis (LAD), four sampling techniques (brush, swab, scrape and adhesive tape strip) were used to study the distribution of yeasts in various body sites and these results were compared with those from five cases of atopic dermatitis and those of 10 normal dogs. Malassezia was frequently isolated from lesional and non-lesional skin and haircoat, footpads, nails and mucous membranes from dogs with either LAD or atopic dermatitis, although, generally, more Malassezia organisms were isolated from LAD cases. In normal dogs, Malassezia was most frequently recovered from the ear canal and the perianal skin. Candida was isolated frequently from dogs with LAD, but only a single isolate of this yeast was found in the other two groups. Fungal hyphae and pseudohyphae, probably Candida albicans, could be detected in samples collected from the nails and footpads of dogs with LAD. Both Malassezia and Candida could be isolated using all four sampling techniques. The MacKenzie (toothbrush) technique and adhesive tape strip cultures proved simple methods for the semiquantitative evaluation of yeasts. The high recovery rate of Malassezia and Candida from dogs with LAD is probably related to immune dysfunction, particularly T-cell dysfunction, known to be present in these dogs. C albicans infection may in part be responsible for the pathogenic changes of the nails and footpads commonly seen in cases of LAD. PMID- 11440399 TI - Haemolytic anaemia and exercise intolerance due to phosphofructokinase deficiency in related springer spaniels. AB - Phosphofructokinase (PFK) deficiency is an autosomal recessive inherited disorder in dogs causing haemolytic crises and exertional myopathy. The clinical signs may be confused with those of recurrent immune-mediated haemolytic anaemia. The deficiency has been commonly observed in field trial (working) English springer spaniels (ESSPs), but also in the conformation line of ESSPs in the USA over the past two decades. This report documents the first family of ESSPs found with PFK deficiency in Europe. Two related adult ESSPs in Denmark had intermittent signs of pigmenturia after exercise (hunting) and had evidence of a regenerative haemolytic anaemia. Based upon DNA sequencing data, both dogs had the previously described nonsense point mutation in the muscle-type PFK gene (delta2228G-->A). Study of 17 related family members using a simple and accurate PFK-DNA test revealed one additional PFK-deficient dog (with minor exercise intolerance), nine carriers and seven normal (or 'clear') ESSPs. Recently, the authors have also identified PFK carriers and affected ESSPs in the UK. Screening for PFK deficiency is recommended for ESSPs with suspicious clinical signs and before using any for field trials or breeding in order to prevent the further spread of this hereditary disorder. PMID- 11440400 TI - Partial bladder resection in a bitch with urinary retention following surgical excision of a vaginal leiomyoma. AB - Ovariohysterectomy and surgical excision of a vaginal leiomyoma were performed in a bitch exhibiting continuous bleeding from the vulva. Following excision of the tumour, urine retention and urinary incontinence developed. Treatment using drugs and an indwelling catheter was unsuccessful in correcting the disorders and chronic cystitis developed after long-standing urine stagnation in the bladder. The authors resorted to undertaking partial cystectomy in order to correct the condition, which resulted in reduced urine retention and improved micturition. PMID- 11440401 TI - Traction alopecia with vasculitis in an Old English sheepdog. AB - A seven-year-old entire male Old English sheepdog was presented with a well circumscribed, completely alopecic area on the top of its head, located where a rubber band had been used for several years. A punch biopsy was taken from the centre of the lesion and histological examination revealed cell-poor, interface mural folliculitis with follicular atrophy and vasculitis. A diagnosis of traction alopecia was made. Oral pentoxyfilline, at 400 mg twice daily, was prescribed for two months, with a slight improvement. To the authors' knowledge, this is the first report of canine traction alopecia accompanied by vascular damage. This vascular damage may represent the pathomechanism of this type of alopecia in the dog. PMID- 11440402 TI - Spinal cord astrocytoma in a cat. AB - A nine-year-old neutered male domestic shorthaired cat with a history of spinal pain and progressive hindlimb dysfunction was presented to the Norwegian School of Veterinary Science. Following neurological and myelographic examination, an intramedullary mass affecting several lumbar spinal cord segments was diagnosed. A neoplastic lesion was suspected and a poor prognosis was given. On postmortem examination, the spinal cord was found to be dorsally flattened from the 12th thoracic vertebra to the fifth lumbar vertebra and severely thickened with a dorsal cleft from the fifth to the seventh lumbar vertebra. Histologically, the tumour was diagnosed as an anaplastic astrocytoma. PMID- 11440403 TI - Outcome in breast cancer. PMID- 11440404 TI - Prognostic implications of p53 and bcl-2 expression in 108 women with stage two breast cancer. AB - BACKGROUND: The mortality and morbidity of patients with breast cancer can vary even between individuals with similar histological stage at diagnosis. Identification of those individuals with prognostically poorer tumours is an essential prerequisite in planning adjuvant therapies. Some prognostic indices of tumour size, grade, oestrogen receptor status and nodal status are well established. AIM: The aim of this study was to examine the prognostic role of information relating to proto-oncogene and tumour suppressor gene expression. METHODS: 108 women with stage II breast cancer were studied. Tumour expression of p53 and bcl-2 were scored and then correlated with recurrence and mortality. RESULTS: We have shown that individuals poorly expressing bcl-2 in their tumours have a poorer disease-free and overall survival than those who express bcl-2. When p53 was strongly expressed, it was associated with poorer disease-free and overall survival. CONCLUSION: The profiling of individual tumour genetic expression of proto-oncogenes may allow for more specific identification of patients at higher risk of recurrence in breast cancer. PMID- 11440405 TI - Self-perceived health status of Irish breast cancer survivors. AB - BACKGROUND: There is a paucity of data on the health status of breast cancer survivors. AIMS: To measure the self-perceived health status of a cohort of breast cancer survivors by means of the SF-36 questionnaire. METHODS: The questionnaire was administered to 120 survey participants. Findings were compared with those of an external comparison group. RESULTS: The health status of the survey participants was found to be significantly better than that of the comparison group on four of the eight SF-36 dimensions--social functioning, mental health, vitality and general health perceptions. CONCLUSIONS: The self perceived health status of breast cancer survivors merits further research in the Irish context. Inclusion of the SF-36 questionnaire as an outcome measure is recommended. PMID- 11440406 TI - Where do patients with cancer die in Belfast? AB - BACKGROUND: Most patients with cancer prefer to die at home but the majority die in institutions. AIM: To determine place of death for patients with cancer in Belfast, to examine changes over time and identify factors associated with place of death. METHODS: A survey of deaths registered in Belfast over a six-month period for 1977, 1987 and 1997 identified patients dying from cancer. Epidemiological data included age, gender, malignancy, social class, marital status, area of residence and place of death. RESULTS: Home deaths fell from 35% in 1977 to 28% in 1997. Hospital deaths fell from 50% in 1977 to 40% in 1987 rising to 42% in 1997. Hospice deaths rose from 13% in 1977 to 25% in 1987 falling to 23% in 1997. There was an association between place of death and age, marital status, type of cancer and area of residence, but not with social class or gender. CONCLUSION: The majority of people fail to achieve a home death. Resources need to be targeted to those most at risk of an institutional death; females, the elderly, the unmarried, those with haematological malignancies and residents of South Belfast. PMID- 11440407 TI - Six-year follow-up after Helicobacter pylori eradication in peptic ulcer disease. AB - BACKGROUND: The longterm outlook after Helicobacter pylori (H. pylori) eradication in peptic ulcer disease is unclear. AIM: This study documents H. pylori recurrence, dyspeptic symptoms and anti-secretory therapy in peptic ulcer patients six years or more after H. pylori eradication. METHODS: Peptic ulcer patients with H. pylori eradication between 1990 and 1992 were included. Infection recurrence was diagnosed by 13-carbon urea breath test (UBT). Dyspeptic symptoms and anti-secretory therapy use were assessed by questionnaire. RESULTS: Sixty-one patients completed the study protocol. Mean follow-up after eradication was 6.1 years (range 4.8-8.3). Four patients had a positive UBT. H. pylori recurrence rate was 6.6% or 0.02% per patient per year. Forty-two patients (69%) had dyspeptic symptoms. Heartburn and belching were more common than pain (p<0.001). All four patients with H. pylori recurrence had symptoms compared with 38 of 57 H. pylori-negative patients (p>0.05). Ten of 61 patients (16.4%) were taking anti-secretory therapy and their dyspepsia scores were higher. CONCLUSIONS: Despite a low H. pylori recurrence rate, longterm dyspeptic symptoms were common in peptic ulcer patients after H. pylori eradication. The symptoms are mainly reflux in type and require anti-secretory therapy in only a minority of patients. PMID- 11440408 TI - Clinical outcome after acquisition of Burkholderia cepacia in patients with cystic fibrosis. AB - BACKGROUND: Respiratory disease is the major cause of morbidity and mortality in cystic fibrosis (CF). The significance of Burkholderia cepacia (B. cepacia) in the pathogenesis of lung disease in CF is debated, but its exact role remains unclear. AIM: To assess the impact of respiratory tract colonisation with B. cepacia in patients with CF by measuring changes in pulmonary function and body mass index (BMI). METHODS: Three groups of patients were defined based on sputum culture isolates: Group 1 were B. cepacia and Pseudomonas aeruginosa (P. aeruginosa) positive patients; Group 2 were P. aeruginosa positive; and Group 3 were colonised with neither organism. Forced expiratory volume (FEV) and BMI were measured annually from 1987 to 1995 and the year of acquisition of P. aeruginosa or B. cepacia was recorded. RESULTS: The mean annual decrease in FEV1 was significantly different in all three groups: Group 1, -5.4 (5.1)%; Group 2, -3.9 (6.5)%; and Group 3, -1.6 (1.0)%, (p<0.05). The mean percentage decrease in FEV1 of a sub-group of Group 1 patients where the B. cepacia acquisition date was known was 6.1% per year versus 1.55% in Group 2 patients (p<0.05). CONCLUSIONS: Acquisition of B. cepacia may be a cause of, rather than a marker for, a decrease in pulmonary function. PMID- 11440409 TI - Blood-borne infections in Dublin's opiate users. AB - BACKGROUND: Injecting drug users are at high risk of acquiring blood-borne infections. Ireland has had a harm reduction policy of methadone maintenance and needle exchange since 1992. AIM: To estimate prevalence of hepatitis B, hepatitis C and HIV infection and appropriate uptake of hepatitis B vaccine in methadone attendees and to make recommendations for a simple record-based surveillance system. METHOD: Retrospective study of 138 client records for evidence of laboratory tests or test results for blood-borne viruses and appropriate immunisation against hepatitis B. RESULTS: A total of 60% of clients had evidence of one or more laboratory tests in their notes. Of those tested for individual viruses, 5.1% were positive for hepatitis B surface antigen, 78.8% had antibodies to hepatitis C and 16.7% were HIV positive. Nearly two-thirds of clients had no evidence of vaccination or information on prior immunity in their records. CONCLUSIONS: A standardised written protocol for screening for blood-borne viruses and for immunisation against hepatitis B in methadone service attendees was clearly needed, and was subsequently introduced by the Eastern Region Health Authority. PMID- 11440410 TI - Opiate-related deaths in Dublin. AB - BACKGROUND: Intravenous drug misuse, principally heroin, occurs primarily in the Greater Dublin area. Methadone maintenance treatment has been an important part of the response to opiate misuse in Dublin since 1992. AIMS: To determine the number of opiate-related deaths in Dublin City and County during 1999, to establish the number of methadone-related deaths and determine the proportion of deaths associated with methadone prescribed according to guidelines. METHODS: A retrospective review of all coroners' inquest files in Dublin City and County during 1999 was undertaken. RESULTS: There were 84 opiate-related deaths. Seventy eight (92.9%) were male. Seventy-three (86.9%) had two or more drugs identified toxicologically. There were 45 methadone-related deaths, of which 15 (33.3%) were receiving methadone prescribed according to guidelines. CONCLUSION: Opiate related deaths occur primarily in males in the 25-34 year age group and are associated with a high level of polydrug use. Diverted methadone accounted for the majority of deaths involving PMID- 11440411 TI - The research abstract: worth getting it right. AB - BACKGROUND: Scientific merit and clarity are critical in evaluation of quality in research. We hypothesised that avoidable errors of presentation adversely impact on abstract selection for scientific meetings. AIM: To prospectively evaluate compliance with abstract guidelines among abstracts submitted to a national surgical scientific meeting. METHODS: Compliance of all submitted abstracts with 13 instructions to authors was compared using ANOVA and Chi-squared tests. Results are expressed as mean (standard deviation, range). RESULTS: Of 45 abstracts submitted, only 8 (17%) complied with all guidelines. Rejected abstracts were less concise than accepted abstracts (280.5 +/- 73.8 words vs. 244.2 +/- 42.5; p=0.006) and were more likely to be rejected (chi2 = 8.67, 1 df, p<0.05). There was no significant difference between the number of errors in accepted (1.6 [1.43, 0-4]) versus rejected (2.4 [1.87, 0-7], ANOVA; p=0.217) abstracts. All late submissions (30%) were rejected. Nine abstracts (20%) contained statistical errors or omissions. CONCLUSIONS: Succinct presentation may reflect clarity of focus or increased writing experience. Reviewers favour concise abstracts. Concise presentation and timely submission are easily achieved and increase the likelihood of research acceptance for scientific meetings. PMID- 11440412 TI - The effect of the anaesthetic agent isoflurane on the rate of neutrophil apoptosis in vitro. AB - BACKGROUND: Volatile anaesthetic agents influence neutrophil function, and potentially, the inflammatory response to surgery. AIM: The objective of this study was to determine the effect of isoflurane (1-4%) on human polymorphonuclear neutrophil apoptosis in vitro. METHODS: Venous blood from 12 healthy volunteers was exposed to 0, 1, and 4% isoflurane delivered via a 14G Wallace flexihub internal jugular cannula, at a fresh gas flow of 0.51/min for 5 minutes. Isolated neutrophils were assessed for apoptosis at 1, 12, and 24 hours in culture using dual staining with annexin V-FITC and propidium iodide (Annexin-V FITC assay). Data were analysed using paired, one-tailed Student's t-tests. p<0.05 was considered significant. RESULTS: At 1 hour apoptosis was inhibited in the 1% (5.1 [6.8]%; p=0.017) and 4% (4.8 [4.5]%; p=0.008) isoflurane groups compared to control (11.3 [6.9]%). At 12 and 24 hours, a dose-dependent inhibition of apoptosis was demonstrated, i.e. 4% > 1% > 0%. CONCLUSION: Human neutrophil apoptosis is inhibited in a concentration-dependent manner in vitro by isoflurane in clinical concentrations. PMID- 11440413 TI - The financial cost to patients of visiting a GP in Ireland: a cross-sectional survey. AB - BACKGROUND: Future general practice funding is much debated but public opinion has not previously been reported. AIM: To survey public opinion on existing fee structures in general practice and on an insurance-based primary care scheme that would entitle members to free GP services at the point of use, upon payment of an annual premium. METHODS: An interviewer administered questionnaire-based cross sectional survey in four locations: Dublin north city; Dublin south city; Dublin suburban; and a rural town centre. RESULTS: Of 384 people surveyed, 83% had visited a GP in the previous year, and 79% usually paid for consultations. The median fee was IEP20.00. Almost 50% felt that this was excessive. Of those who did not have to pay, 91% expressed willingness to pay (median IEP15.00). Sixty two per cent had medical insurance and 83% said they would be willing to join an insurance-based primary care scheme. The amount people would pay to join this scheme exceeded what interviewees were currently paying for general practice services. CONCLUSIONS: There is dissatisfaction with existing GP fees and support for a scheme entitling free primary care. The amount of money people are willing to pay exceeds their current annual expenditure on GP services. PMID- 11440414 TI - Predictors of first hip fracture and mortality post fracture in older women. AB - BACKGROUND: Hip fracture causes significant morbidity and mortality in older women. AIM: To document factors contributing to the risk of hip fracture in older women and to assess the effect of hip fracture on subsequent mortality. METHODS: Case-control study of 89 women with hip fracture and 89 controls, with two-year follow-up. Singh index and bone mineral density were calculated. RESULTS: Osteoporotic indices did not differ significantly between cases and controls. Significant predictors of risk were sleeping tablets, perception of health as fair/poor and a lower mental status score. Patients were 3.57 times more likely to die in the first year after fracture, with no difference between the groups in year two. After adjustment, hip fracture did not remain significantly associated with mortality. Inability to walk 100 yards alone prior to fracture and lower social class were significantly associated with mortality at 12 months. Age alone was significantly associated at 12-24 months. CONCLUSIONS: Factors related to falls and fracture may be more discriminatory predictors of hip fracture risk than osteoporosis in older females. Medications for sleep should be prescribed with caution. Hip fracture may have an independent effect on one year mortality, this effect is not seen in the second year. PMID- 11440415 TI - The genetics of osteoporosis: vitamin D receptor gene polymorphisms and circulating osteocalcin in healthy Irish adults. AB - BACKGROUND: A relationship between bone turnover and the vitamin D receptor (VDR) genotype is reported in Australian subjects of UK-Irish descent. There is conflicting evidence of linkage between the VDR genotype and differences in bone mineral density. No such study has been carried out on a resident Irish population. AIMS: This study examined the relationship between serum osteocalcin (a marker of bone turnover) and VDR genotype in Irish adults. METHODS: One hundred and eighteen healthy Irish adults aged between 19 and 67 were recruited and had two fasting blood samples taken. One sample was analysed for osteocalcin by enzyme immunoassay. The other was used to isolate genomic DNA and determine VDR genotype. RESULTS: Tt was the most prevalent VDR genotype (53%) and tt (15%) the least prevalent in this group of healthy Irish adults. The tt VDR genotype was associated with significantly higher levels of serum osteocalcin (29% and 40%) than the Tt or TT VDR genotypes. CONCLUSIONS: These findings suggest that healthy Irish adults with the tt VDR genotype have higher rates of bone turnover than those with Tt or TTVDR genotypes. They may have a higher risk of low bone mineral density and osteoporosis in later life. PMID- 11440416 TI - Exercise and osteoporosis. AB - Osteoporosis is a systemic skeletal disease characterised by low bone density and micro architectural deterioration of bone tissue with a consequent increase in bone fragility and susceptibility to fracture. It is a silent disease. It is the result of the negative balance between bone formation and bone resorption, i.e. more bone is lost than is formed. It is the most common bone disease worldwide and is now a major health problem. Bones require a normal level of systemic hormones, adequate caloric intake, particularly protein, calcium and vitamin D and regular weight-bearing exercise throughout life. A large bone mass early in life protects against osteoporosis. Peak bone mass is determined by sex, heredity family history, race, diet and exercise. Sixty per cent of bone growth occurs during adolescence. Moderate exercise protects against osteoporosis, but too little or excessive exercise may cause osteoporosis. PMID- 11440417 TI - Advances in radiation technology can improve survival and quality of life for cancer patients. PMID- 11440418 TI - Coma secondary to aciclovir neurotoxicity. PMID- 11440419 TI - Adult intussusception: a report of five cases. PMID- 11440420 TI - Medicine and early Irish law. PMID- 11440421 TI - Management of Boerhaave's syndrome in a regional setting. PMID- 11440422 TI - Hiring consumer-providers: barriers and alternative solutions. AB - The hiring of consumers as providers of mental health services has steadily increased over the last decade. This article, based on the literature and two round table discussions, explores three prevalent barriers (i.e., dual relationships, role conflict, and confidentiality) and proposes alternative solutions to each. PMID- 11440423 TI - Chronicity reconsidered: improving person-environment fit through a consumer-run service. AB - In the past, the term "chronic" referred to people who had serious mental illness and who typically received long-term care in a state mental hospital. Although this term recently has fallen out of favor, we resurrect the term here, not to revive a demeaning euphemism, but rather to redefine it as the result of a poor person-environment fit between the complex and challenging needs of those with serious psychiatric disorders and a community-based service system that often is ill-equipped to treat them. Previous research indicates that recurrent acute hospitalizations and an inability to establish or maintain tenure in the community may be due to a disconnection from community-based services and supports, social isolation, and demoralization. One promising approach to addressing these issues is that of peer support. To illustrate the potential utility of peer support in improving person-environment fit and decreasing the chronicity of the subsample of people who continue to have difficulty in establishing viable footholds in the community, we describe a peer support-based program, the Welcome Basket, developed, staffed, and managed entirely by mental health consumers. Preliminary analyses that evaluate Welcome Basket's effectiveness are included, and we discuss the implications of these data for future research and program development in this area. PMID- 11440424 TI - Assessing support needs of caregivers of persons with dementia: who wants what? AB - This paper documents the use and interest in support strategies such as telephone support, newsletters, and computer services, among caregivers of all ages. Data obtained from telephone interviews with community living caregivers of persons with dementia (n = 148) showed substantial interest in the use of these types of services. Using a theoretical framework, this paper identifies caregiver and care receiver characteristics associated with those interested in the utilization of these support strategies. The implications of these findings for program planning and future research are discussed. PMID- 11440425 TI - Alternative modalities of help within socio-political and ethnic minorities: self help among Arabs living in Israel. AB - The present paper deals with alternative modalities of help within sociopolitical and ethnic minorities. It explores the phenomenon of self-help as one modality of help within the Arab population living in Israel. The analysis is based on four research studies, which incorporate the perspectives of lay people, professionals, self-help group members, and students in the helping professions, regarding definitional issues associated with the concept of self-help and attitudes toward such a modality of help. Findings obtained from the four studies reported here suggest that a natural fit exists between the Arab culture and such a helping modality as self-help organizations. Based on the findings, practical recommendations are outlined for use in the area of social work with socio political and ethnic minorities. PMID- 11440426 TI - PTSD among Bosnian refugees: a survey of providers' knowledge, attitudes and service patterns. AB - The objective of this study was to investigate providers' knowledge, attitudes and service provision patterns for Bosnian refugees with PTSD and to consider the overall implications for trauma-related mental health services. A survey instrument was developed and administered to 30 randomly selected service providers working with Bosnian refugees in Chicago. This sample includes primary medical care providers (n = 7), community social service providers (n = 12), and mental health service providers (n = 11). Results showed that knowledge about PTSD is highest in mental health service providers, as is amount of previous training and education about PTSD. Less than half of all providers systematically assess for PTSD, and standardized instruments for PTSD are rarely used. The top three overall recommendations for services were ranked as: 1. Psychotherapy; 2. No intervention; 3. Psychiatric evaluation. In general, less than half of providers hold attitudes toward refugees as suffering from illnesses or mental disorders, or that "something" is wrong with them. Likewise, only half of the providers reported providing education to refugees and their families about the possible mental health consequences of trauma. In conclusion, assessment, intervention and educational activities of providers are not consistent with literature documenting that PTSD is highly prevalent in refugee populations. New training programs, model development, and research initiatives are needed to address the needs of refugees. PMID- 11440427 TI - Determinants of anti-psychotic medication compliance in a multicultural population. AB - The efficacy of medication in controlling or ameliorating symptoms of psychotic conditions is well established in clinical trials, but medication effectiveness in mental health services is considerably affected by the extent to which clients actually comply to their prescribed medication. This study aimed to assess the relative influence of demographic, medication, social support and relationship variables on perceived compliance with medication in a sample of clients with serious mental illness from diverse ethnic backgrounds. It also aimed to explore whether compliance differed by matching clients with a case manager from the same ethno-linguistic background. Data on medication compliance, administration route, medication dose, sex, age, impairment of thought processes, mood abnormality, ability to manage money, acceptance of illness (insight), social network size, alcohol and drug use, impulse control, and cooperation with staff were provided by case managers for 168 clients from diverse ethnic backgrounds. Multiple regression analysis was used to examine the predictors of medication compliance. The main predictors of greater compliance were greater general cooperation with staff, better insight, and matching clients with a case manager from the same ethnic background. PMID- 11440428 TI - Mental health training in emergency homeless shelters. AB - The prevalence of mental illness among homeless persons points to the importance of providing mental health training to emergency shelter staff. The authors report on their own work and argue that such training offers the potential to significantly improve shelter staffs ability to respond to the needs of shelter residents with mental illness, and to the behavioral problems some of these individuals may pose for shelter operation. Mental health care providers should take into consideration organizational dynamics when planning and implementing such training. PMID- 11440429 TI - The Alexandrian Library: crucible of a renaissance. AB - At the end of the 4th century BC, the Macedonian-Greek armies of Alexander the Great swept across Asia from Egypt to the Indus River, redefining political boundaries within that vast territory at a time when important cultural changes were also taking place in the Greek world. New literary forms were beginning to emerge from the classical literature, which was then the subject of scholarly investigation. There was growing curiosity about the physical world and mathematics. Aristotle and his contemporaries were redefining scholarship at a time when Alexander was redefining the political sphere. These remarkable transformations converged in Alexandria, which became the center of a new intellectual universe. The first Ptolemaic rulers founded two unique institutions -the Alexandrian Library and the Mouseion--and the Library became the crucible within which the Hellenistic renaissance was forged. PMID- 11440430 TI - Surgery on anterior foramen magnum meningiomas using a conventional posterior suboccipital approach: a report on an experience with 17 cases. AB - OBJECTIVE: The advantages of a posterior "conventional" suboccipital approach with a midline incision over lateral, anterolateral, and anterior approaches to anteriorly placed foramen magnum meningiomas are discussed. METHODS: From 1991 to March 2000, 17 patients with foramen magnum meningiomas arising from the anterior or anterolateral rim of the foramen magnum underwent operations in the Department of Neurosurgery at King Edward Memorial Hospital and Seth G.S. Medical College. All patients were operated on in a semi-sitting position by use of a conventional suboccipital approach with a midline incision and extension of the craniectomy laterally toward the side of the tumor up to the occipital condyle. RESULTS: The patients ranged in age from 17 to 72 years, and the tumors ranged in size from 2.1 to 3.8 cm. The intradural vertebral artery was at least partially encased on one side in eight patients and on both sides in two patients. The brainstem was displaced predominantly posteriorly in each patient. A partial condylar resection was performed in two cases to enhance the exposure. Total tumor resection was achieved in 14 patients, and a subtotal resection of the tumor was performed in the other 3 patients. In one patient, a small part of the tumor was missed inadvertently, and in the other two patients, part of the tumor in relation to the vertebral artery and posterior inferior cerebellar artery was deliberately left behind. After surgery, one patient developed exaggerated lower cranial nerve weakness. There was no significant postoperative complication in the remainder of the patients, and their conditions improved after surgery. The average length of follow-up is 43 months, and there has been no recurrence of the tumor or growth of the residual tumor. CONCLUSION: From our experience, we conclude that a large majority of anterior foramen magnum meningiomas can be excised with a lateral suboccipital approach and meticulous microsurgical techniques. PMID- 11440431 TI - Adjunctive use of endoscopy during posterior fossa surgery to treat cranial neuropathies. AB - OBJECTIVE: The objective of this study was to determine the utility and safety of rigid endoscopy as an adjunct during posterior fossa surgery to treat cranial neuropathies. METHODS: A suboccipital craniotomy was performed for 19 patients with non-neoplastic processes involving the Vth, VIIth, and/or VIIIth cranial nerves. Ten patients with trigeminal neuralgia (n = 8), hemifacial spasm (n = 1), or intractable tinnitus (n = 1) underwent primarily microvascular decompression procedures. One patient with geniculate neuralgia underwent nervus intermedius sectioning combined with microvascular decompression. Eight patients underwent unilateral vestibular nerve neurectomies for treatment of Meniere's disease. A 0- or 30-degree rigid endoscope was used in conjunction with the standard microscopic approach for all procedures. RESULTS: All patients experienced resolution or significant improvement of their preoperative symptoms after posterior fossa surgery. The endoscope allowed improved definition of anatomic neurovascular relationships without the need for significant cerebellar or brainstem retraction. Cleavage planes between the cochlear and vestibular nerves entering the internal auditory canal and sites of vascular compression could not be microscopically observed for several patients; however, endoscopic identification was possible for all patients. There were no complications related to the use of the endoscope. CONCLUSION: The rigid endoscope can be used safely during posterior fossa surgery to treat cranial neuropathies, and it allows improved observation of the cranial nerves, nerve cleavage planes, and vascular anatomic features without significant cerebellar or brainstem retraction. PMID- 11440432 TI - New technique for application of fibrin sealant: rubbing method devised to prevent cerebrospinal fluid leakage from dura mater sites repaired with expanded polytetrafluoroethylene surgical membranes. AB - OBJECTIVE: The effectiveness of a strong, pressure-tight method for fibrin sealant application was experimentally and clinically evaluated, using expanded polytetrafluoroethylene (ePTFE) surgical membranes. METHODS: Two application methods were examined in two groups. For Group 1, 1 ml each of Solutions A and B was simply sprayed over the surface (spray method). For Group 2, a small amount of Solution A was rubbed into the sutured site and then Solutions A and B were sprayed over the surface (rubbing method). In the experimental study, a moment pressure elevation test and a continuous stepwise pressure elevation test were performed for both groups. The clinical study was conducted using 350 patients undergoing craniotomies. The ePTFE surgical membrane was used for 180 patients but not the other 170 patients. RESULTS: In the experimental study, the bursting pressures were 51.4 +/- 13.0 cm H2O for Group 1 and 129 +/- 35.0 cm H2O for Group 2 in the moment pressure elevation test and 27.0 +/- 6.7 cm H2O and 100 +/- 31.9 cm H2O, respectively, in the continuous stepwise pressure elevation test. The sealing effect was significantly greater for Group 2 than for Group 1 in both tests (P < 0.001). In the clinical study using ePTFE surgical membranes, the rubbing method significantly reduced the incidence of postoperative cerebrospinal fluid leakage (1.4%), compared with the spray method (18.3%), when external decompression was not concomitantly used (P < 0.01). CONCLUSION: The sealing effect of fibrin sealant on cerebrospinal fluid leakage was significantly greater with the rubbing method than with the spray method. The strong sealing effect obtained with the rubbing method is thought to result from firm linkage between the ePTFE surgical membrane and the fibrin sealant and also from fibrin being formed even in needle holes and spaces in the sutured site. PMID- 11440433 TI - Evidence of increased endothelial cell turnover in brain arteriovenous malformations. AB - OBJECTIVE: We hypothesized that human brain arteriovenous malformations (BAVMs) are nonstatic vascular lesions with active angiogenesis or vascular remodeling. To test this hypothesis, we assessed endothelial cell turnover in BAVMs. METHODS: We identified nonresting endothelial cells by use of immunohistochemistry for the Ki-67 antigen. From archived paraffin blocks, we selected BAVM vessels without intravascular thrombosis or embolic material in areas nonadjacent to the nidus edge. For controls, we used 50- to 100-microm diameter cortical vessels from temporal lobe cortex removed for epilepsy treatment. The Ki-67 index was calculated as a percentage of Ki-67-positive endothelial cells. The data were analyzed by the nonparametric Mann-Whitney test and reported as mean +/- standard deviation. RESULTS: Thirty-seven specimens that met the above criteria were selected. There were 26 +/- 15 vessels counted in each BAVM specimen versus 18 +/ 5 in each control cortex (n = 5). The mean Ki-67 index was higher for BAVM vessels than control cortical vessels (0.7 +/- 0.6 versus 0.1 +/- 0.2%; P = 0.005), which represented an approximately seven-fold increase in the number of nonresting endothelial cells. In the BAVM group, there was a trend for younger patients to have a wider variation and higher Ki-67 index than older patients; no trend was evident in the control group. CONCLUSION: Compared with control vessels, BAVM vessels have higher endothelial cell turnover, which suggests the presence of active angiogenesis or vascular remodeling in BAVMs. PMID- 11440434 TI - Genetic evaluation of lipoprotein(a) in intracranial aneurysm disease. AB - OBJECTIVE: Elevations in serum lipoprotein(a) [Lp(a)] levels have been reported in intracranial aneurysm (IA) disease. Our aim was to investigate a genetic basis for this observation. METHODS: We performed a comparative analysis of size polymorphisms at two loci (kringle 4 [K4] and TTTTA pentanucleotide [PN] repeats) within the apolipoprotein(a) gene on Chromosome 6q26-27 among patients with sporadic IAs (n = 50), members of three IA families (n = 50), and control subjects (n = 50). RESULTS: There was no significant difference in mean Lp(a) levels between patients with sporadic IAs and control subjects, but IA family members exhibited a more than twofold elevation in mean Lp(a) levels, compared with control subjects (29.2 versus 12.9 mg %). Inverse relationships between K4/PN numbers and serum Lp(a) levels were demonstrated; genotype frequencies did not differ significantly from a Hardy-Weinberg equilibrium or from published frequencies for other Caucasian populations. We detected no difference in mean K4 and PN genotypic indices between patients with IAs and control subjects (9.3 and 16.92 versus 9.0 and 16.92, respectively), but IA families did exhibit a lower mean K4 genotypic index (7.7), compared with control subjects. Superficial analysis of family pedigrees revealed no suggestion of linkage between K4/PN genotypes and IA disease. CONCLUSION: The previously described elevation in Lp(a) levels among patients with sporadic IAs might be explained by an acute-phase response. Crude Lp(a) measurements might provide a useful predictive test for familial IA disease, but with the disadvantage of low specificity. The possibility of linkage of familial IA disease to a particular apolipoprotein(a) isoform size range has not been eliminated. PMID- 11440435 TI - Regrowth of the rostral spinal axons into the caudal ventral roots through a collagen tube implanted into hemisected adult rat spinal cord. AB - OBJECTIVE: A collagen tube was used to guide axonal regrowth from the spinal cord to the periphery to contribute to improvement of paralysis after lower thoracic spinal cord injury. METHODS: The spinal cords of adult male Sprague-Dawley rats were lesioned by removing the left hemicord from T12 to 5 mm below this level and additionally sectioning all left lumbar ventral roots. In experimental animals (n = 9), a collagen tube was inserted into this gap, spanning the rostral hemisected cord to the caudal sectioned lumbar ventral roots (gap, 7 mm). In control animals (n = 6), no treatment was performed. RESULTS: Six months after surgery, the return of some tension and resistance of the paralyzed hindlimb muscles was observed in all experimental rats except the untreated controls. Nine months postoperatively, muscle action potentials were recorded from the target muscles of the experimental animals while electrostimulating the tissue continuity within the collagen tube. Horseradish peroxidase retrograde labeling showed that the neurons in the rostral cord near the implantation site regrew into the reconnected lumbar ventral roots. Histological examination indicated numerous myelinated axons in the reconnected root pathways and newly formed endplates in the target muscles. No axonal regeneration was found in the control rats. CONCLUSION: These results indicate that the rostral spinal axons can regrow into the caudal sectioned and reconnected ventral roots through a collagen tube, thus innervating the denervated peripheral targets in adult rats after spinal cord injury. This surgical repair model also provides a means for testing the use of trophic factors that may further promote axonal regeneration. PMID- 11440436 TI - Temporal clustering of hemorrhages from untreated cavernous malformations of the central nervous system. AB - OBJECTIVE: Hemorrhages from cerebral cavernous malformations (CMs) sometimes seem to occur in closely spaced "clusters" interspersed with long hemorrhage-free intervals. Clustering of hemorrhages could affect retrospective assessments of radiosurgery efficacy in prevention of CM rehemorrhage. However, this empirical observation had not been tested quantitatively. To test whether CM hemorrhages tend to cluster, we reviewed pretreatment rebleeding rates after a first symptomatic hemorrhage in CM patients who later underwent surgery or radiosurgery. METHODS: We performed a retrospective review of 141 patients with CMs who presented with clinically overt hemorrhage, and who subsequently underwent surgery or proton beam radiosurgery during an 18-year period. Statistical models were used to analyze all events per person and identify potential variation in rebleeding risk with time after a previous hemorrhage. RESULTS: Sixty-three of 141 patients experienced a second hemorrhage before treatment; 16 had additional hemorrhages. Five hundred thirty-eight patient years elapsed between first hemorrhages and treatment. The cumulative incidence of a second hemorrhage after the first CM hemorrhage was 14% after 1 year and 56% after 5 years. During the first 2.5 years after a hemorrhage, the monthly rehemorrhage hazard was 2%. The risk then decreased spontaneously to less than 1% per month, which represents a 2.4-fold decline (P < 0.001). Rehemorrhage rates were higher in younger patients (P < 0.01), but not in females or in patients with deep lesions. Shorter intervals between successive hemorrhages did not predict higher subsequent rehemorrhage risk. CONCLUSION: The rehemorrhage rate from untreated CMs is high initially, and it decreases 2 to 3 years after a previous hemorrhage. This hazard pattern generates the observed temporal clustering of hemorrhages from untreated CMs. PMID- 11440437 TI - Detrimental effects of systemic hyperthermia on locomotor function and histopathological outcome after traumatic spinal cord injury in the rat. AB - OBJECTIVE: Posttraumatic hyperthermia has been demonstrated to worsen neurological outcome in models of brain injury. The purpose of this study was to examine the effects of systemic hyperthermia on locomotor and morphological outcome measures after traumatic spinal cord injury (SCI) in the rat. METHODS: After a T10 laminectomy, spinal cord contusions were produced from a height of 12.5 mm onto exposed cords (NYU Impactor; New York University Neurosurgery Laboratory, New York, NY) in adult rats that were divided into three groups. Group 1 (n = 9) underwent whole body hyperthermia (rectal temperature, 39.5 degrees C) 30 minutes postinjury for 4 hours, Group 2 (n = 8) underwent normothermia (rectal temperature, 37 degrees C) 30 minutes postinjury for 4 hours, and Group 3 (n = 10) underwent traumatic SCI with no postinjury thermal treatment. Twice-weekly assessments of locomotor function were made during a 6 week survival period using the Basso-Beattie-Breshnahan locomotor rating scale. Forty-four days after injury, animals were perfused, and their spinal cords serially sectioned. Sections were stained with hematoxylin, eosin, and Luxol fast blue for histopathological analysis. The percentage of tissue damage was quantitatively determined by using computer-aided image analysis. RESULTS: The results showed that 4 hours of postinjury hyperthermia significantly worsened locomotor outcome (final Basso-Beattie-Breshnahan scores were 9.7 +/- 0.3 [Group 1] versus 10.8 +/- 0.4 [Group 2] versus 11.3 +/- 0.3 [Group 3]) and led to an increase in the percentage of tissue damage (32.9 + 3.2% [Group 1] versus 22.3 +/ 2.8% [Group 3]). CONCLUSION: These data suggest that complications of SCI (e.g., fever, infection) leading to an elevation of systemic temperature may add to the severity of secondary injury associated with traumatic SCI and significantly affect neurological outcome. PMID- 11440438 TI - Preferential conditions for and possible mechanisms of induction of ischemic tolerance by repeated hyperbaric oxygenation in gerbil hippocampus. AB - OBJECTIVE: We reported previously that repeated hyperbaric oxygenation (HBO) as pretreatment induced ischemic tolerance in the gerbil hippocampus. This study was conducted to determine the preferential conditions for induction of ischemic tolerance by HBO and the mechanism of this induction through immunohistochemical analysis of Bcl-2, Bax, and manganese superoxide dismutase expression. METHODS: Five-minute forebrain ischemia was produced in gerbils after pretreatment with 2 atmospheres absolute (ATA) HBO once every other day for one, three, or five sessions, 2 ATA hyperbaric air once every other day for five sessions, or 3 ATA HBO once daily for 10 sessions. Histological examinations were then performed. Two days after pretreatment with 2 ATA HBO once every other day for five sessions or with 3 ATA HBO once daily for 10 sessions, sections were analyzed immunohistochemically. RESULTS: Pretreatment with 2 ATA HBO once every other day for three or five sessions induced ischemic tolerance; however, pretreatment with 2 ATA HBO for one session, 2 ATA hyperbaric air once every other day for five sessions, or 3 ATA HBO once daily for 10 sessions did not. Pretreatment with 2 ATA HBO once every other day for five sessions, but not with 3 ATA HBO once daily for 10 sessions, significantly increased Bcl-2 and manganese superoxide dismutase immunoreactivity in the CA1 sector. CONCLUSION: These results suggest that protection against mitochondrial alterations after ischemia through manganese superoxide dismutase and/or Bcl-2 expression may be related to induction of ischemic tolerance by repeated HBO pretreatment. PMID- 11440439 TI - Tumor Fas (APO-1/CD95) up-regulation results in increased apoptosis and survival times for rats with intracranial malignant gliomas. AB - OBJECTIVE: The cellular "death" receptor Fas has been proposed to be a potential specific target for anti-glioma therapy. However, little is known regarding the effects of Fas expression on glioma viability in vivo. The goal of this study was to clarify the relationships among Fas expression, apoptosis, and survival rates for high-grade astrocytomas. METHODS: Fas expression was measured in several human glioblastoma multiforme cell lines and a malignant rat glioma cell line (36B10), before and after Fas up-regulation by gene transfer. Expression was correlated with the degree of Fas-mediated cytotoxicity and apoptosis induced after Fas activation. Subsequently, rats underwent intracranial implantation of either wild-type or genetically altered 36B10 cell lines, for study of the effects of Fas up-regulation on survival rates. RESULTS: Low levels of cell surface Fas expression in glioblastomas multiforme were correlated with their limited susceptibility to Fas-mediated cytotoxicity. Through Fas receptor up regulation, relationships among increased Fas expression, Fas-mediated cytotoxicity, and apoptosis were demonstrated. The percentage of cells undergoing apoptosis after exposure to a Fas ligand-producing cell line increased from 4% in the sham-transfected line (36B10-) to 27% in the Fas-transfected line (36B10 Fas). After intracranial implantation of these tumors into rats, the median survival time increased significantly from 14 days (36B10 and 36B10-) to 24.5 days (36B10-Fas), which represents a 75% increase in the survival time for the greater Fas-expressing group (P = 0.0005). CONCLUSION: It seems that the overall low rate of apoptosis in high-grade astrocytomas is related to low levels of cell surface Fas expression. With increases in cellular Fas expression, rates of Fas mediated apoptosis and survival rates were increased. PMID- 11440440 TI - Caspase-9 transduction overrides the resistance mechanism against p53-mediated apoptosis in U-87MG glioma cells. AB - OBJECTIVE: Conflicting reports have been published with regard to the relationship between the efficacy of p53 gene therapy and the p53 status of gliomas. In this study, we evaluated whether U-87MG glioma cells harboring wild type p53 and U251 and U-373MG glioma cells harboring mutated p53 demonstrate different sensitivities to p53-induced apoptosis. In addition, we tested whether transduction of Bax or caspase-9, which are downstream components of p53-induced apoptosis, can override the resistance mechanism of U-87MG cells to apoptosis. METHODS: We transduced U-87MG, U251, and U-373MG glioma cells with p53, Bax, or caspase-9 genes via adenovirus (Adv) vectors, to induce the same level of respective proteins, and evaluated the degree of apoptosis. RESULTS: U-87MG cells were highly resistant to Adv for p53 (Adv-p53)-mediated apoptosis, whereas U251 and U-373 cells underwent extensive apoptosis after Adv-p53 infection. In U-87MG cells, the elevation of Bax and Fas was not as marked as that observed in U251 and U-373MG cells after Adv-p53 infection. Endogenous expression of Bcl-XL and Bcl-2 in U-87MG cells was greater than that in U251 and U-373MG cells. U-87MG cells were more resistant to Bax-mediated apoptosis than were U251 or U-373MG cells. In contrast, U-87MG cells were more sensitive to caspase-9-mediated apoptosis than were U251 or U-373MG cells, suggesting that transduction of caspase-9 may override the resistance mechanism of U-87MG to p53-mediated apoptosis. CONCLUSION: These results demonstrate that proapoptotic function induced by p53 transduction in U-87MG cells was repressed at several steps and that induction of caspase-9 may circumvent this resistance mechanism. PMID- 11440441 TI - The Valley of the Golden Mummies. PMID- 11440442 TI - The museum's mummies: an inside view. AB - We applied medical and scientific methodology in a study of the mummies in the Metropolitan Museum of Art in New York, New York, by computed tomographic scanning and other radiographic methods. These noninvasive procedures allow us to "unwrap" these mummies without unwrapping them. This is, in effect, technology transfer of routine diagnostic techniques used in medicine to Egyptology. After this noninvasive information-gathering procedure, the mummies are preserved intact for possible future investigations that may be more sophisticated and more informative. The data are presented and put into perspective by a review of pertinent literature. PMID- 11440443 TI - Multiple intracranial aneurysms as delayed complications of an atrial myxoma: case report. AB - OBJECTIVE AND IMPORTANCE: Atrial myxomas are rare cardiac tumors that may cause neurological complications; however, delayed neurological events after total tumor resection are rare. In this report, we present a patient who developed transient cerebral ischemic attacks and was found to have multiple intracranial aneurysms 5 years after successful resection of her atrial myxoma. At the time of myxoma resection, there were no neurological symptoms; at the time of presentation with transient ischemic attacks, there was no evidence of atrial recurrence. CLINICAL PRESENTATION: A 32-year-old woman presented with five episodes of right arm and face paresthesia, each lasting 15 to 20 minutes, 5 years after successful resection of her atrial myxoma. Clopidogrel bisulfate therapy was initiated, with resolution of her symptoms. Angiography revealed multiple, peripherally located, fusiform cerebral aneurysms. INTERVENTION: A left frontal craniotomy for resection and biopsy of one of the aneurysms was performed, to establish the diagnosis. Pathological analysis of the biopsied aneurysm provided evidence of direct atrial myxoma invasion and occlusion of the cerebral blood vessel. CONCLUSION: Neurological symptoms may accompany or lead to the diagnosis of atrial myxoma. Rarely, as in this case, myxomatous aneurysms may develop years after definitive treatment of the primary tumor. Patients who have undergone successful resection of a left atrial myxoma may be at risk for delayed cerebral ischemia associated with aneurysm development, and this phenomenon must be considered for patients with neurological symptoms who present even years after myxoma removal. The natural history, pathophysiological features, and treatment dilemma of these aneurysms are discussed. PMID- 11440444 TI - Patent foramen ovale as a possible risk factor for cryptogenic brain abscess: report of two cases. AB - OBJECTIVE AND IMPORTANCE: Patent foramen ovale (PFO) has been suggested as a potential source of paradoxical embolism. A higher prevalence of PFO in ischemic stroke of unexplained cause has been recognized. Brain abscesses are commonly associated with a contiguous focus of infection, hematogenous spread from a distant focus, or cranial trauma. However, no predisposing factors, including a distant focus with unknown cause, are identified in approximately 15 to 30% of reported cases. CLINICAL PRESENTATION: We encountered two patients with brain abscess presumably caused by dental infections. Both patients displayed PFO, through which right-to-left atrial contrast shunting was revealed by transesophageal echocardiography. Although the radiological location of the abscesses suggested hematogenous spread as a cause, the patients had no arteriovenous shunting other than the PFO, despite exhaustive investigations for a potential infectious route. The patients displayed no definite focal orofacial inflammatory signs during the postoperative course despite diagnosis of pyorrhea alveolaris or periodontitis. INTERVENTION: In Patient 1, the abscess was aspirated stereotactically, and in Patient 2, the abscess disappeared radiologically after high-dose antibiotic treatment. CONCLUSION: The mechanism of brain abscess formation putatively related to PFO should be different from that related to common dental sepsis. Analysis of these cases suggested that infectious embolism from a latent or even identifiable focus through the PFO may be an underrecognized cause of brain abscess, in contrast to simple seeding of the brain via transit of the infecting bacteria through the valveless emissary veins. PMID- 11440445 TI - Posterior fossa decompression and clot evacuation for fourth ventricle hemorrhage after aneurysmal rupture: case report. AB - OBJECTIVE AND IMPORTANCE: Massive intraventricular hemorrhage due to aneurysmal rupture is associated with a dismal prognosis. An intraventricular clot causing fourth ventricle dilation can cause compression to the brainstem similar to other posterior fossa masses such as cerebellar hemorrhage or infarction. The presence of fourth ventricle dilation carries a very high risk of death within 48 hours. Neither ventricular drainage nor fibrinolytic infusion has been successful in eliminating clots of the fourth ventricle. Posterior fossa decompression and direct evacuation of the clot could have good results in relieving brainstem compression caused by the clot. CLINICAL PRESENTATION: A 45-year-old woman was admitted to our intensive care unit after experiencing an aneurysmal subarachnoid hemorrhage. The neurological examination at admission revealed that she was in Grade V according to the World Federation of Neurological Surgeons grading system, but brainstem reflexes were present. Computed tomographic scanning revealed a massive intraventricular hemorrhage, with fourth ventricle dilation caused by an intraventricular clot. Bilateral external ventricular drains were placed to relieve elevated intracranial pressure. Cerebral angiography revealed a 1-cm basilar tip aneurysm, which was embolized with Guglielmi detachable coils (Boston Scientific, Boston, MA) during the same procedure. INTERVENTION: Given the patient's poor neurological condition, it was decided that brainstem compression should be relieved. A posterior fossa decompressive craniectomy was performed immediately after coil therapy, with direct evacuation of the intraventricular clot. The patient experienced a clear improvement in the level of consciousness and has achieved a good neurological result at early follow-up. CONCLUSION: Dilation of the fourth ventricle by an intraventricular clot is a sign of brainstem compression that can be relieved by posterior fossa decompression and direct clot evacuation. PMID- 11440446 TI - Arachnoid cyst of the craniocervical junction: case report. AB - OBJECTIVE AND IMPORTANCE: An arachnoid cyst at the craniocervical junction presenting with obstructive hydrocephalus as a result of blockage of the outflow of the fourth ventricle is described. This is a very rare anatomic site, with only five other cases described in the literature. CLINICAL PRESENTATION: A 37 year-old woman presented with a 9-month history of severe neck pain, persistent vomiting, visual disturbances, and numbness of the nose, cheek, and lips. She had severe bilateral papilledema on ophthalmoscopy. Magnetic resonance imaging revealed a midline cystic lesion extending down to C2. INTERVENTION: The patient underwent posterior fossa craniectomy and excision of the arachnoid cyst. She made a full recovery and was asymptomatic at follow-up examination. CONCLUSION: The symptomatology of these rare craniocervical arachnoid cysts and their development are discussed. PMID- 11440447 TI - Microsurgical removal of a petrous apex meningioma after stereotactic radiation: technical case report. AB - OBJECTIVE AND IMPORTANCE: Stereotactic radiation is increasingly advocated as a primary treatment option for benign cranial base lesions. The clinical course of the patient reported herein raises questions regarding the rationale for initiation of radiotherapy to a petrous apex meningioma before microsurgery. CLINICAL PRESENTATION: We report a 50-year-old woman who experienced medically refractory trigeminal pain. She was diagnosed with a meningioma around the petrous apex and treated by fractionated stereotactic radiation. After a short period of alleviation accompanied by hypesthesia, the pain returned in a previously unknown and violent fashion. INTERVENTION: Complete tumor removal through a retrosigmoid intradural suprameatal approach resulted in immediate and permanent pain cessation. CONCLUSION: Radiotherapy should be withheld for benign and accessible tumors of the cranial base until the option of radical microsurgical treatment has been explored. PMID- 11440448 TI - How to make a three-dimensional surgical illustration of cerebral vessels with a personal computer: technical note. AB - OBJECTIVE: To introduce a method of making three-dimensional (3-D) illustrations of cerebral vessels with a personal computer. METHODS: We generated the 3-D images from only two views of conventional angiograms (anteroposterior and lateral views) with a personal computer, two commercial software programs, and an image scanner. RESULTS: This method was applied to 10 consecutive cases of cerebral aneurysms. 3-D images were made in a few hours, and they contributed to assessing vascular structures of an aneurysm complex. CONCLUSION: This method is useful for discussing surgical strategies preoperatively. Our method may be used in any facility where conventional angiography, personal computers, and 3-D software are available. PMID- 11440449 TI - Carotid balloon occlusion for large and giant aneurysms: evaluation of a new test occlusion protocol. PMID- 11440450 TI - Management of severe traumatic brain injury by decompressive craniectomy. PMID- 11440451 TI - The epidemiology of brain arteriovenous malformations. PMID- 11440452 TI - Intraarterial recombinant tissue plasminogen activator for ischemic stroke: an accelerated dosing regimen. PMID- 11440453 TI - Double aneurysms at distal basilar artery: report of nine cases. PMID- 11440454 TI - Cerebrovascular surgery: past, present, and future. PMID- 11440455 TI - The neuron doctrine, the mind, and the Arctic. PMID- 11440456 TI - The nature and fate of punctate (type IV) cavernous malformations. AB - OBJECTIVE: Four types of cavernous malformations (Types I-IV) have been described on the basis of their magnetic resonance imaging (MRI) appearance. The nature of the Type IV cavernous malformation is unclear. It has been suggested that these small lesions, which are well observed only on gradient echo MRI scans, are capillary telangiectasias. We sought to understand the relationship of Type IV cavernous malformations to the other cavernous malformation subtypes. METHODS: We examined serial MRI scans obtained between 1987 and 2000 from 68 patients with more than 228 cavernous malformations. Sixteen patients harbored Type IV cavernous malformations (total, >114 Type IV lesions). Spin echo T1-weighted, T2 weighted, proton density, and (when available) gradient echo MRI scans were reviewed. Cavernous malformations that met the Zabramski criteria for Type IV (poorly observed on T1- and T2-weighted images) were reviewed in serial scans from individual patients to characterize their radiographic behavior over time. RESULTS: Type IV cavernous malformations were best observed on gradient echo images and have an MRI appearance distinct from capillary telangiectasias. Proton density images demonstrate more Type IV lesions than T1- and T2-weighted images, but far fewer Type IV lesions than gradient echo images. When observed on T1- and T2-weighted images, Type IV cavernous malformations are generally punctate and hypointense. These lesions rarely enhance with gadolinium. Four of the Type IV cavernous malformations observed serially evolved into Type I and Type II cavernous malformations, for an approximate rate of progression of 0.05 per patient year. CONCLUSION: Although most Type IV cavernous malformations remain stable over time, a small subset of these lesions progress into Types I and II cavernous malformations. PMID- 11440457 TI - Measurement of nitric oxide and brain tissue oxygen tension in patients after severe subarachnoid hemorrhage. AB - OBJECTIVE: Nitric oxide (NO), one of the most powerful endogenous vasodilators, is thought to play a major role in the development of delayed vasospasm in patients with subarachnoid hemorrhage (SAH). However, the role of the production of cerebral NO in patients with SAH is not known. In other SAH studies, NO metabolites such as nitrite and nitrate have been demonstrated to be decreased in cerebrospinal fluid and in plasma. METHODS: In this study, a microdialysis probe was used, along with a multiparameter sensor, to measure NO metabolites, brain tissue oxygen tension, brain tissue carbon dioxide tension, and pH in the cortex of patients with severe SAH who were at risk for developing secondary brain damage and vasospasm. NO metabolites, glucose, and lactate were analyzed in the dialysates to determine the time course of NO metabolite changes and to test the interrelationship between the analytes and clinical variables. RESULTS: Brain tissue oxygen tension was strongly correlated to dialysate nitrate and nitrite (r2 = 0.326; P < 0.001); however, no correlation was noted between brain tissue oxygen tension and NO metabolites in cerebrospinal fluid (r2 = 0.018; P = 0.734). No significant correlation between NO production, brain tissue carbon dioxide tension, and dialysate glucose and lactate was observed. CONCLUSION: Cerebral ischemia and compromised substrate delivery are often responsible for high morbidity rates and poor outcomes after SAH. The relationship between brain tissue oxygen and cerebral NO metabolites that we demonstrate suggests that substrate delivery and NO are linked in the pathophysiology of vasospasm after SAH. PMID- 11440458 TI - Intra-arterial third-generation recombinant tissue plasminogen activator (reteplase) for acute ischemic stroke. AB - OBJECTIVE: We prospectively evaluated the safety and recanalization efficacy of intra-arterially administered reteplase, a third-generation recombinant tissue plasminogen activator, for treating ischemic stroke in patients considered poor candidates for intravenously administered alteplase therapy. METHODS: Patients were considered poor candidates for intravenously administered therapy because of severity of neurological deficits, interval from onset of symptoms to presentation of 3 hours or more, or recent major surgery. We administered a maximum total dose of 8 U of reteplase intra-arterially in 1-U increments via superselective catheterization. Adjunctive angioplasty of the occluded artery was performed in seven patients. Angiographic evidence of perfusion and thrombus was graded by use of modified Thrombolysis in Myocardial Infarction (TIMI) criteria. Neurological examinations were performed before and 24 hours and 7 to 10 days after treatment. RESULTS: Sixteen consecutive patients were treated (mean age, 64.1 +/- 16.4 yr; seven were men). Initial National Institutes of Health Stroke Scale scores ranged from 10 to 26. Time from onset of symptoms to treatment ranged from 2 to 9 hours. Occlusion sites were the cervical internal carotid artery (n = 4), intracranial internal carotid artery (n = 4), middle cerebral artery (n = 6), and vertebrobasilar artery (n = 2). Complete or near-complete perfusion (TIMI Grade 3 or 4) was achieved in the arteries in 14 patients (88%), with partial recanalization (TIMI Grade 2) or minimal response (TIMI Grade 1) in the arteries in one patient each. Neurological improvement (defined as decrease of four or more points in National Institutes of Health Stroke Scale score) was observed in 7 (44%) of the 16 patients at 24 hours. Symptomatic intracerebral hemorrhage occurred in one patient; three other patients experienced intracerebral hemorrhages that did not result in neurological worsening. The overall mortality during hospitalization was 56%, related to massive ischemic stroke (n = 7), withdrawal of care at the family's request after the development of aspiration pneumonia and renal failure (n = 1), and a combination of intracerebral hemorrhage and massive ischemic stroke (n = 1). CONCLUSION: In this study, intra-arterially administered reteplase in doses up to 8 U with or without angioplasty resulted in a high rate of recanalization. This strategy should be considered in treating patients considered poor candidates for intravenous thrombolysis. PMID- 11440459 TI - Complications of epilepsy surgery after 654 procedures in Sweden, September 1990 1995: a multicenter study based on the Swedish National Epilepsy Surgery Register. AB - OBJECTIVE: To present the first national multicenter study on complications after epilepsy surgery procedures to include all epilepsy surgery centers in Sweden: Goteborg, Linkoping, Lund, Stockholm, Umea, and Uppsala. METHODS: Every epilepsy surgery procedure in Sweden is reported to the Swedish National Epilepsy Surgery Register. The report includes relevant social and medical items and complications. A complication is defined as minor if it resolves within 3 months and major if it affects activities of daily living and lasts longer than 3 months. Follow-up data are recorded for 2 years after the operation. Intrinsic checkpoints and external revision validate the register data. RESULTS: During the inclusion period (September 1990-December 1995), 654 surgical procedures were performed (age range, 6 mo-67 yr). Of these, 205 were invasive electrode procedures (182 first investigations and 23 reinvestigations), and 449 were therapeutic procedures (375 first operations and 74 reoperations). After invasive electrode procedures, only minor complications were reported (6.3%). For all 449 therapeutic procedures (including reoperations), minor complications were reported in 8.9% and major complications in 3.1 %. Only one major complication was reported in a patient under the age of 35 years. CONCLUSION: This is the first national multicenter study on complications after epilepsy surgery. It demonstrates that these procedures as performed at six different centers in Sweden are safe, but also that risk is related to age. In patients younger than age 35 years, the risk for a major complication after invasive subdural strip electrode investigation and epilepsy surgery is low. PMID- 11440460 TI - High-dose trigeminal neuralgia radiosurgery associated with increased risk of trigeminal nerve dysfunction. AB - OBJECTIVE: Stereotactic radiosurgery is being used with more frequency in the management of patients with trigeminal neuralgia. To improve facial pain outcomes, many centers have increased the prescribed radiation dose to the trigeminal nerve. METHODS: Between April 1997 and December 1999, 68 patients underwent radiosurgery for trigeminal neuralgia with use of the Leksell gamma knife (Elekta Instruments, Norcross, GA) and a single 4-mm isocenter of radiation. Twenty-seven patients (40%) received 70 Gy (low dose) of irradiation and 41 patients (60%) received 90 Gy (high dose). The groups were similar with regard to age, sex, duration of pain, number of prior surgeries, and preexisting trigeminal deficits. The primary facial pain outcomes for analysis were excellent (pain-free, no medications) and good (pain-free, reduced medications). The mean length of follow-up after radiosurgery was 14.4 months (range, 2-36 mo). RESULTS: At last follow-up examination, 11 (41%) of the 27 patients with low-dose radiosurgery remained pain-free compared with 25 (61%) of the 41 patients with high-dose radiosurgery (P = 0.17). Additional surgery was performed in 12 low dose patients (44%) and 8 high-dose patients (20%) (P = 0.05). High-dose radiosurgery was associated with an increased rate of permanent trigeminal nerve dysfunction (54% versus 15%, P = 0.003). Bothersome dysesthesias occurred in 13 high-dose patients (32%), whereas only 1 low-dose patient had this complication (P = 0.01). Three high-dose patients (8%) developed corneal numbness after radiosurgery. Pain recurred with more frequency in patients not developing trigeminal nerve dysfunction after radiosurgery (9 of 22 patients, 41 %) compared with those who sustained facial numbness, paresthesias, or dysesthesias (4 of 27 patients, 15%); however, the difference was not statistically significant (P = 0.08). CONCLUSION: Higher doses of radiation may correlate with better facial pain outcomes after radiosurgery for trigeminal neuralgia. However, the incidence of significant trigeminal nerve dysfunction is markedly increased after radiosurgery for patients receiving high-dose radiosurgery. Because of the nonselective nature of this ablative technique, dose prescription should be limited to less than 90 Gy. PMID- 11440461 TI - C1-C2 transarticular screw fixation for atlantoaxial instability: a 6-year experience. AB - OBJECTIVE: We review a 6-year, single-center experience using the technique of C1 C2 transarticular screw fixation for atlantoaxial instability in 75 consecutive operations. METHODS: The study group was composed of 43 men and 32 women, with a mean age of 44 years (range, 8-76 yr). Each patient had documented atlantoaxial instability. In 28 patients (37%), atlantoaxial instability was a result of trauma; in 22 patients, (29%), it was a result of rheumatoid arthritis; in 16 patients (21%), it was a result of prior surgery; and in 9 patients (12%), it was a result of congenital abnormalities. All patients underwent stabilization with C1-C2 transfacetal screws and a posterior interspinous construct. Nine patients had unilateral screws placed. Postoperatively, the patients were maintained in a rigid cervical orthosis for a mean of 11 weeks (range, 8-15 wk); five patients were immobilized with halo fixation for a mean of 13 weeks (range, 10-16 wk). The mean follow-up period was 2.4 years (range, 1-5.5 yr). RESULTS: Osseous fusion was documented in 72 patients (96%). There were no hardware failures; however, three patients developed pseudarthrosis. Two superficial wound infections (one at the graft site and one at the cervical incision site) required antibiotic therapy. Four patients had transient suboccipital hypesthesia. No instances of an errant screw, dural laceration, or injury to the vertebral artery, spinal cord, or hypoglossal nerve were noted. CONCLUSION: C1-C2 transarticular screw fixation supplemented with an interspinous construct yielded a 96% fusion rate, with a low incidence of complications. We attribute our successful outcomes to careful preoperative assessment and meticulous surgical technique. PMID- 11440462 TI - C1-C2 transarticular screw fixation: technical aspects. AB - OBJECTIVE: I review posterior atlantoaxial fusion with transarticular screw fixation, including indications, complications, and operative technique, emphasizing my experience. METHODS: The indications for C1-C2 transarticular screw fixation include traumatic injuries to the atlantoaxial complex, instability resulting from inflammatory disease (rheumatoid arthritis), and congenital abnormalities (os odontoideum). All patients underwent stabilization using cannulated C1-C2 transfacetal screws by the method described by Magerl. Supplemental interspinous fusion with bicortical autologous iliac crest graft and titanium cable was used to restore the posterior tension band by use of the method described by Sonntag's group. Preoperatively, all patients underwent imaging with plain radiographs, magnetic resonance imaging, and axial computed tomography. Patients were maintained in a rigid cervical orthosis postoperatively. RESULTS: Measures used to improve safety and efficacy include patient positioning, fluoroscopic guidance, preoperative magnetic resonance imaging, axial computed tomography, and open reduction of C1-C2 subluxation before screw passage. In this series of 75 patients, fusion was obtained in 72 patients (96%). There were no instances of vertebral artery injury, errant screw placement, instrumentation failure, dural laceration, spinal cord injury, or hypoglossal nerve injury. CONCLUSION: C1-C2 transarticular screw fixation with a posterior tension band construct provides excellent fusion rates with few perioperative complications. Preoperative imaging and meticulous surgical technique improve outcomes. PMID- 11440463 TI - Modeling of retraction and resection for intraoperative updating of images. AB - OBJECTIVE: Intraoperative tissue deformation that occurs during the course of neurosurgical procedures may compromise patient-to-image registration, which is essential for image guidance. A new approach to account for brain shift, using computational methods driven by sparsely available operating room (OR) data, has been augmented with techniques for modeling tissue retraction and resection. METHODS: Modeling strategies to arbitrarily place and move an intracranial retractor and to excise designated tissue volumes have been implemented within a computationally tractable framework. To illustrate these developments, a surgical case example, which uses OR data and the preoperative neuroanatomic image volume of the patient to generate a highly resolved, heterogeneous, finite-element model, is presented. Surgical procedures involving the retraction of tissue and the resection of a left frontoparietal tumor were simulated computationally, and the simulations were used to update the preoperative image volume to represent the dynamic OR environment. RESULTS: Retraction and resection techniques are demonstrated to accurately reflect intraoperative events, thus providing an approach for near-real-time image-updating in the OR. Information regarding subsurface deformation and, in particular, changing tumor margins is presented. Some of the current limitations of the model, with respect to specific tissue mechanical responses, are highlighted. CONCLUSION: The results presented demonstrate that complex surgical events such as tissue retraction and resection can be incorporated intraoperatively into the model-updating process for brain shift compensation in high-resolution preoperative images. PMID- 11440464 TI - Three-dimensional visualization of the pyramidal tract in a neuronavigation system during brain tumor surgery: first experiences and technical note. AB - OBJECTIVE: To integrate spatial three-dimensional information concerning the pyramidal tracts into a customized system for frameless neuronavigation during brain tumor surgery. METHODS: Four consecutive patients with intracranial tumors in eloquent areas underwent diffusion-weighted and anatomic magnetic resonance imaging studies within 48 hours before surgery. Diffusion-weighted datasets were merged with anatomic data for navigation purposes. The pyramidal tracts were segmented and reconstructed for three-dimensional visualization. The reconstruction results, together with the fused-image dataset, were available during surgery in the environment of a customized neuronavigation system. RESULTS: In all four patients, the combination of reconstructed data and fused images was a helpful additional source of information concerning the tumor seat and topographical interaction with the pyramidal tract. In two patients, intraoperative motor cortex stimulation verified the tumor seat with regard to the precentral gyrus. CONCLUSION: Diffusion-weighted magnetic resonance imaging allows individual estimation of large fiber tracts applicable as important information in intraoperative neuronavigation and in planning brain tumor resection. A three-dimensional representation of fibers associated with the pyramidal tract during brain tumor surgery is feasible with the presented technique and is a helpful adjunct for the neurosurgeon. The main drawbacks include the length of time required for the segmentation procedure, the lack of direct intraoperative control of the pyramidal tract position, and brain shift. However, mapping of large fiber tracts and its intraoperative use for neuronavigation have the potential to increase the safety of neurosurgical procedures and to reduce surgical morbidity. PMID- 11440465 TI - Transsphenoidal approaches for the extracapsular resection of midline suprasellar and anterior cranial base lesions. AB - OBJECTIVE: The transsphenoidal approach is an effective method for treating tumors contained within the sella or extending into the suprasellar cistern. The technique of tumor dissection is predicated on preservation of the integrity of the diaphragma, i.e., intracapsular removal. Gross total extracapsular dissection may, however, be accomplished either by using a standard approach to the pituitary fossa or by extending the exposure to include removal of a portion of the planum sphenoidale and division of the superior intercavernous sinus. METHODS: Included in this series were 14 patients with parasellar or sellar tumors with extension into the anterior fossa and/or suprasellar cistern. For 4 of 14 patients (29%), extracapsular access was gained by broaching the tumor capsule from within the pituitary fossa. For the remaining 10 of 14 patients (71%), the dura of the floor of the sella and the planum sphenoidale was exposed, using neuronavigation to verify the limits of bony dissection; extracapsular tumor resection was performed using the operating microscope and endoscopy as indicated. The dural defect was repaired with abdominal fat, the sellar floor and planum sphenoidale were reconstructed, and in selected cases a lumbar drain was placed. RESULTS: Seven of 14 tumors (50%) were craniopharyngiomas, 3 of 14 (21%) were pituitary adenomas, and 2 of 14 (14%) were meningiomas. There was one case of lymphocytic hypophysitis and one yolk sac tumor. Gross total resection was possible in 11 of 14 cases (79%). Immediate postoperative visual function worsened in 2 of 14 cases (14%), improved in 3 of 14 cases (21%), and was stable in the remainder of cases. Postoperatively, 2 of 14 patients (14%) developed bacterial meningitis. Overt postoperative cerebrospinal fluid rhinorrhea was not observed. CONCLUSION: Gross total extracapsular resection of midline suprasellar tumors via a transsphenoidal approach is possible but is associated with a higher risk of complications than is standard transsphenoidal surgery. PMID- 11440466 TI - Recent developments in non-excitable cell calcium entry. AB - Influx of calcium into cells following stimulation of cell surface receptors is a key process controlling cellular activity. However, despite intensive research, there is still no consensus on precisely how calcium entry is controlled in electrically no n-excitable cells. In particular, the regulation of depletion activated or 'capacitative' calcium entry continues to be a focus of debate. Work published in the last 2 years has lent new impetus to the so-called 'conformational coupling' theory, although evidence for the existence of soluble messengers between the ER and the plasma membrane also continues to appear. In addition, there remains disagreement on whether intra-store [Ca(2+)] has to fall below a threshold before Ca(2+)entry is activated. A further major question is the identity of the putative depletion-operated Ca(2+)channel or channels. Here discussion has largely focussed on whether homologue(s) of the Drosophila TRP ('Transient Receptor Potential') protein is/are the elusive channel, or at least a part of it. Finally, it remains possible that Ca(2+)entry mechanisms other than depletion-activated channels may be important in agonist-evoked Ca(2+)influx. This commentary summarizes recent developments in the field, and highlights both current debates and critical unsolved questions. PMID- 11440467 TI - Inositol 1,4,5-trisphosphate-independent calcium signalling by platelet-derived growth factor in the human SH-SY5Y neuroblastoma cell. AB - In adherent SH-SY5Y human neuroblastoma cells, activation of G-protein-coupled muscarinic M3 receptors evoked a biphasic elevation of both intracellular [Ca(2+)] ([Ca(2+)]i) and inositol-1,4,5-trisphosphate (D-Ins(1,4,5)P3) mass. In both cases, temporal profiles consisted of rapid transient elevations followed by a decline to a lower, yet sustained level. In contrast, platelet-derived growth factor (PDGF), a receptor tyrosine kinase agonist acting via PDGF receptor b chains in these cells, elicited a slow and transient elevation of [Ca(2+)]i that returned to basal levels within 5 to 10 min with no evidence of inositol phosphate generation. Full responses for either receptor type required intracellular and extracellular Ca(2+) and mobilization of a shared thapsigargin sensitive intracellular Ca(2+) store. Strategies that affected the ability of D Ins(1,4,5)P3 to interact with the Ins(1,4,5)P3-receptor demonstrated an Ins(1,4,5)P3-dependency of the muscarinic receptor-mediated elevation of [Ca(2+)]i but showed that PDGF-mediated elevations of [Ca(2+)]i are Ins(1,4,5)P3 independent in these cells. PMID- 11440468 TI - Ca(2+)-regulated nitric oxide generation in rabbit parotid acinar cells. AB - In rabbit parotid acinar cells, the muscarinic cholinergic agonist methacholine induced an increase in the intracellular Ca(2+) concentration and provoked nitric oxide (NO) generation. Ca(2+)-mobilizing reagents such as thapsigargin and the Ca(2+) ionophore A23187 mimicked the effect of methacholine on NO generation. Methacholine-induced NO generation was inhibited by the removal of extracellular Ca(2+). Immunoblot analysis indicated that the antibody against the neuronal type of nitric oxide synthase (NOS) cross-reacted with NOS in the cytosol of rabbit parotid gland cells. Immunofluorescence testing showed that neuronal NOS is present in the cytosol of acinar cells but less in the ductal cells. NOS was purified approximately 8100-fold from the cytosolic fraction of rabbit parotid glands by chromatography on Sephacryl S-200, DEAE-Sephacel, and 29,59-ADP Sepharose. The purified NOS was a NADPH- and tetrahydroxybiopterin-dependent enzyme and was activated by Ca(2+) within the physiological range in the presence of calmodulin. These results suggest that NO is generated by the activation of the neuronal type of NOS, which is regulated in rabbit parotid acinar cells by the increase in intracellular Ca(2+) levels induced by the activation of muscarinic receptors. PMID- 11440469 TI - 2-aminoethoxydiphenyl borate reveals heterogeneity in receptor-activated Ca(2+) discharge and store-operated Ca(2+) influx. AB - We have investigated Ca(2+) release and receptor- and store-operated Ca(2+) influxes in Chinese hamster ovary-K1 (CHO) cells, SH-SY5Y human neuroblastoma cells and RBL-1 rat basophilic leukemia cells using Fura-2 and patch-clamp measurements. Ca(2+) release and subsequent Ni(2+)-sensitive, store-operated influx were induced by thapsigargin and stimulation of G protein-coupled receptors. The alleged noncompetitive IP3 receptor inhibitor,2 aminoethoxydiphenyl borate (2-APB) rapidly blocked a major part of the secondary influx response in CHO cells in a reversible manner. It also reduced Mn(2+) influx in response to thapsigargin. Inhibition of Ca(2+) release was also seen but this was less complete, slower in onset, less reversible, and required higher concentration of 2-APB. In RBL-1 cells, I(CRAC) activity was rapidly blocked by extracellular 2-APB whereas intracellular 2-APB was less effective. Store operated Ca(2+) influxes were only partially blocked by 2-APB. In SH-SY5Y cells, Ca(2+) influxes were insensitive to 2-APB. Ca(2+) release in RBL-1 cells was partially sensitive but in SH-SY5Y cells the release was totally resistant to 2 APB. The results suggest, that 2-APB (1) may inhibit distinct subtypes of IP3 receptors with different sensitivity, and (2) that independently of this, it also inhibits some store-operated Ca(2+) channels via a direct, extracellular action. PMID- 11440470 TI - Dynamics of intracellular calcium in hair cells isolated from the semicircular canal of the frog. AB - Changes in cytosolic free Ca(2+) concentration ([Ca(2+)]i) were monitored optically in hair cells mechanically isolated from frog semicircular canals using the membrane-impermeant form of the Ca(2+)-selective dye Oregon Green 488 BAPTA-1 (OG, 100 microM). Cells stimulated by depolarization under whole-cell voltage clamp conditions revealed Ca(2+) entry at selected sites (hotspots) located mostly in the lower (synaptic) half of the cell body. [Ca(2+)]i at individual hotspots rose with a time constant tau1 approximately 70 ms and decayed with a bi exponential time-course (tau2 approximately 160, tau3 approximately 2500 ms) following a 160 ms depolarization to -20 mV. With repeated stimulation [Ca(2+)]i underwent independent amplitude changes at distinct hotspots, suggesting that the underlying Ca(2+) channel clusters can be regulated differentially by intracellular signalling pathways. Block by nifedipine indicated that the L-type Ca(2+)channels are distributed at different densities in distinct hotspots. No diffusion barrier other than the nuclear region was found in the cytosol, so that, during a prolonged depolarization (lasting up to 1s), Ca(2+) was able to reach the cell apical ciliated pole. The effective Ca(2+) diffusion constant, measured from the progression of Ca(2+) wavefronts in the cytosol, was approximately 57 microm(2)/s. Our results indicate that in these hair cells, buffered diffusion of Ca(2+) proceeds evenly from the source point to the cell interior and is dominated by the diffusion constant of the endogenous mobile buffers. PMID- 11440471 TI - Reduced store-operated Ca(2+) currents in rat basophilic leukaemia cells cultured under serum-free conditions. AB - Influx of Ca(2+) represents an important regulatory signal in the process of cell proliferation. However, little is known about how Ca(2+) entry changes during the cell-cycle. Patch-clamp experiments and microfluorimetry show that store-operated Ca(2+) entry was substantially reduced in rat basophilic leukaemia cells cultured for 24h under serum-free conditions. Likewise, retinoic acid treatment blocked Ca(2+) influx activated by store depletion via inositol 1,4,5-trisphosphate. Both procedures are known to arrest cells at the G0/G1 boundary of the cell-cycle and induced a reduction in 5-bromo 2'-deoxyuridine incorporation into DNA. Ca(2+) release from the stores remained unaltered and two types of K(+) currents were not affected in cells after serum starvation. The specific reduction in Ca(2+) entry was not detected when using aphidicolin, 5-fluorouracil or thymidine to synchronise the cell-cycle. These data suggest that store-operated Ca(2+) influx changed during cell-cycle progression which might have important implications for cell growth. PMID- 11440472 TI - Toxicokinetics of atrazine in embryos of the zebrafish (Danio rerio). AB - Atrazine (2-chloro-4-ethylamino-6-isopropylamine-s-triazine) is a widely used selective herbicide. Due to its persistence, it is present in many surface waters, contaminating nontarget organisms such as fish. Teratogeny of atrazine was examined during the first 48 h of zebrafish development (embryo test) and correlated to effects on the microsomal and soluble glutathione S-transferases (m /sGST; EC 2.5.1.18). Atrazine at 4 mg/L disturbed the normal development to long pec stage, at concentrations between 10 and 20 mg/L it caused retardations in organogenesis, a slowdown of movements, and functional disturbances of heart and circulatory system. m- and sGST activities were affected by atrazine in all investigated developmental stages. Atrazine elevated the mGST activity dose dependently up to a concentration of 5 mg/L, and the sGST up to 1 mg/L. Higher atrazine concentrations caused a decrease of GST activity, especially of the sGST of the younger stages, 32 cells and 75% epiboly. This might indicate that the detoxication system itself is affected, which then gives rise to morphological abnormalities during fish development. In vitro tests were performed to purify and characterize GST-formed atrazine metabolites by the use of ESI--HPLC--MS. mGST showed higher conversion rates compared to sGST. PMID- 11440473 TI - QSAR and chemometric approaches for setting water quality objectives for dangerous chemicals. AB - In order to evaluate environmentally safe levels of dangerous chemicals, there is the need for a set of toxicological data on organisms representative of the ecosystems, which is often unavailable or inadequate. In this article, a predictive approach was applied to a set of 125 chemicals (derived from the European priority list in compliance with Directive 76/464/EEC), for which water quality objectives were available. Toxicological data on organisms representative of the aquatic environment (algae, Daphnia, and fish) were taken from the literature or predicted by means of quantitative structure--activity relationships. This provided toxicological data on all three organisms for 97 of 125 chemicals and on at least two organisms (Daphnia and fish) for the whole data set. Principal Component Analysis was applied in order to perform an a priori classification of chemicals based on toxicity data. Then several classification models, based on traditional and nontraditional molecular descriptors, were applied. Classification models gave results in agreement with the a priori classification as well as with the original water quality objectives classification. The behavior of some outliers was explained. The approach described appears to be a useful tool for the preliminary classification of chemicals that are dangerous to the aquatic environment for which toxicological data are inadequate. PMID- 11440474 TI - Micronuclei and other nuclear lesions as genotoxicity indicators in rainbow trout Oncorhynchus mykiss. AB - The induction of micronuclei and other nuclear abnormalities in renal erythrocytes of rainbow trout Oncorhynchus mykiss by six genotoxic compounds is evaluated. Colchicine, mitomycin, cyclophosphamide, acrylamide, methyl methanesulfonate, and N-ethyl-N-nitrosourea were intraperitoneally injected in trout. Our results show that cyclophosphamide induces the formation of micronuclei and also the other nuclear abnormalities; N-ethyl-N-nitrosourea, acrylamide, and colchicine induce only micronuclei; mitomycin-C induces only other nuclear abnormalities but not micronuclei. Methyl-methanesulfonate does not induce nuclear abnormalities in rainbow trout at the dose assayed in this work. The possible genotoxic origin for the different nuclear abnormalities is discussed. PMID- 11440475 TI - Effects of subchronic exposure to aluminium in acidic water on bioenergetics of Atlantic salmon (Salmo salar). AB - Atlantic salmon (Salmo salar) were exposed for 36 days to water from the acidified (pH 5.2) Fossbekk River (Norway) with 50 micro x L(-1) of aluminium (Al) added as AlCl(3), or to circumneutral water (pH 6.6) from Ims River (Norway). Food consumption, cardiac output, swimming activity, and growth rate were measured to determine the effects of sublethal levels of Al in acidic water on the bioenergetics of the fish. Food consumption was reduced in acidic water and Al during the first 12 days, but was not significantly different from the control group during the rest of the exposure. A significant decrease in body weight was concomitant with the reduction in food consumption. The fish also kept losing weight during the rest of the exposure despite the return of their appetite to normal. Cardiac output of the fish exposed to acidic water and Al was not significantly different from the control group after 36 days of exposure. Swimming activity remained significantly elevated throughout the exposure to acidic water and Al. These results indicate that sublethal levels of Al can alter the energy budget of Atlantic salmon living in acidified surface waters. PMID- 11440476 TI - Differential effects of thiobencarb toxicity on growth and photosynthesis of Anabaena variabilis with changes in phosphate level. AB - A thiobencarb dose of 3 mg L(-1) reduced the protein content of Anabaena variabilis, whereas it elevated the carbohydrate content. Measurements of Anabaena growth, photosynthetic activity, and respiration rate revealed that the high dose of phosphate (0.53 mM) exerted no additional toxic effect to thiobencarb toxicity. Recovery of Anabaena cells from the inhibitory effect of thiobencarb occurred immediately after its reculture in herbicide free-medium. Maximum uptake of thiobencarb was associated with high biomass yield. PMID- 11440477 TI - The Acute toxicity of alpha-branched phenylsulfonyl acetates in Photobacterium phosphoreum test. AB - The acute toxicity (15-min EC(50)) of 20 alpha-substituted phenylsulfonyl acetates was measured using the Microtox test with Photobacterium phosphoreum. On the basis of TLSER, quantitative structure-activity relationships (QSARs) were calculated for the inhibition of bioluminescence. With the parameters molecular volume (V(mc)), dipolarity/polarizability (pi*), and the covalent contribution to Lewis basicity (epsilon(b)), a squared correlation coefficient (r(2)(adj)) of 0.868 and standard error of 0.094 for log EC(50) were obtained. By introducing a structural indicator variable (I), the regressive quality was improved (correlation coefficient of 0.948 and standard error of 0.059); the descriptors V(mc), pi*, and the electrostatic basicity contribution (q(-)) were significant. NO(2)-- and -SO(2)--groups are active centers of these compounds. The strong hydrogen bonding effects between the compound active centers and FMNH(2) play an important role in the greater toxicity of these chemicals. PMID- 11440478 TI - The interaction of Prussian blue and dissolved hexacyanoferrate ions with goethite (alpha-FeOOH) studied to assess the chemical stability and physical mobility of Prussian blue in soils. AB - Colloidal Prussian blue and dissolved hexacyanoferrate ions strongly interact with the surface of goethite (alpha-FeOOH) particles. Whereas Prussian blue is deposited on the surface of goethite as a solid phase with all the properties typical of solid Prussian blue, the hexacyanoferrate ions form initially a layer of surface bound hexacyanoferrate ions, which exhibit properties intermediate between those of free hexacyanoferrate ions and hexacyanoferrate ions bound in Prussian blue. The products of interaction of goethite with Prussian blue and hexacyanoferrate ions were studied by voltammetry of immobilized microcrystals, infrared spectrometry, Vis diffuse reflection spectrometry, transmission electron microscopy, and ESR spectroscopy. The kinetics of Prussian blue destruction was measured by solution Vis spectrometry and also by voltammetry of solid microcrystals. The remarkable stability of Prussian blue in soils can be explained by its strong adherence as a solid phase to iron oxides and oxide hydroxides in soils. This prevents the formation of soluble colloids of Prussian blue. It does not prevent the hydrolysis of Prussian blue to iron oxide hydroxides and hexacyanoferrate(II) ions at elevated pH which proceeds at almost the same rate as that of pure Prussian blue colloid. Even when this hydrolysis of Prussian blue occurs, it is the hexacyanoferrate(II) that is effectively adsorbed on the surface of iron oxide hydroxides, again decreasing the physical mobility of hexacyanoferrate. PMID- 11440479 TI - Baia Mare accident--brief ecotoxicological report of Czech experts. AB - On 30 January 2000, following the breach in the tailing dam of the Aurul SA Baia Mare Co., a major spill of about 100,000 m(3) of cyanide and metal-rich liquid waste was released into the river system near Baia Mare in northwest Romania. The pollutants flowed via different tributaries into the Tizsa (Tisa) river and finally through the Danube river into the Black Sea. Along the way pollutants (especially cyanides) caused rapid death of aquatic organisms and animals living close to the poisoned rivers. Following request from the governments of Romania, Hungary, and the Federal Republic of Yugoslavia, United Nations sent international mission experts to the area of accident. The Czech team of four experts took very active part in this mission. Samples of surface water collected by this team 3 weeks after the accident showed high toxicity in the rivers on the Romanian territory. Due to this impact, water from the Szamos river and a large area of the Tisza river in Hungarian territory was still medially toxic. Improvement of this situation was detected only in downstream areas of the Tisza/Tisa river. The high toxicity of surface water from the locality of the Lapos river upstream of the accident indicated the possibility of toxic pollution from other sources in the area. Aquatic sediments were highly toxic only in the Aurul reservoir and its surrounding area. Rapid decline of their toxicity indicated more limited adverse impact of the "Aurul pollution" in comparison with surface water. From ecotoxicological results it is evident that there is an urgent need to start abatement activities in the Baia Mare area because the possibility of future accidents still exists (this hypothesis was proved by further accidents on 10.3.2000 and the beginning of May 2000). PMID- 11440480 TI - Rating systems for pesticide risk classification on different ecosystems. AB - A novel approach is proposed to quantitatively assess the environmental risks associated with the use of plant protection products. Different ranking indexes for the classification of pesticide risk in various environmental systems at different time and space scales have been developed: PRIHS-1 and PRIHS-2 (Pesticide Risk Index for Hypogean Soil Systems), PRIES-1 and PRIES-2 (Pesticide Risk Index for Epygean Soil Systems), and PRISW-1 and PRISW-2 (Pesticide Risk Index for Surface Water Systems). Such indexes identify the risk for each environmental system immediately after a pesticide spraying (PRIHS-1, PRIES-1, and PRISW-1) or in a wider time--space scale scenario (PRIHS-2, PRIES-2, and PRISW-2). Moreover, a general index (ERIP: Environmental Risk Index of Pesticides) was developed for quantifying the comprehensive risk for the environment. The indexes were calibrated by applying to a large number of pesticides for which data were available. The results of the different indexes are compared and the value and limitations of the approach are discussed. PMID- 11440481 TI - Toxicity of organotin compounds to activated sludge. AB - Inhibition of respiration rate of activated sludge heterotrophic microorganisms was determined for tributyltin (TBT), dibutyltin (DBT), monobutyltin (MBT), and triphenyltin (TPhT). TBT, DBT, and TPhT exhibit similar acute toxicity to activated sludge, while MBT is less toxic. The effect of various experimental parameters, such as sludge age, concentrations of suspended solids, and exposure time, on toxicity was investigated. An increase in sludge age or in the concentration of suspended solids reduces the observed inibition. Longer exposure seems to increase the inhibition of TBT, DBT, and TPhT during the first hours of exposure, while later the inhibition remains constant. In the case of MBT, 24 h after exposure, the respiration rate was similar to that of the control biomass. PMID- 11440482 TI - Radon levels in abandoned metalliferous mines, Devon, southwest England. AB - Radon levels underground in two abandoned mines in Devon, United Kingdom, are reported and analyzed. Extremely high levels have been noted, 7,100,000 Bq m(-3) being the highest level recorded. This is approximately 89 times higher than the highest published radon level for caves and mines in Devon and Cornwall, England, which is 80,000 Bq m(-3). These levels have significant health implications for both casual and occupational mine explorers. PMID- 11440483 TI - Use of the ciliated protozoan Tetrahymena pyriformis for the assessment of toxicity and quantitative structure--activity relationships of xenobiotics: comparison with the Microtox test. AB - Cytotoxicity and quantitative structure-activity relationships of 13 inorganic and 21 organic substances were determined using three bioassays performed on the ciliated protozoan Tetrahymena pyriformis and the luminescent bacterium Vibrio fischeri. The best concordance of toxicity results was observed between the T. pyriformis FDA--esterase activity and population growth inhibition tests for the organic compounds. The sensitivity of these two assays is compared with that of the Microtox test. The T. pyriformis FDA test showed a high sensitivity is most cases. The aim of the current research was to determine whether the relative toxicity of metal ions and organic molecules, with these three bioassays, was predictable using three ion characteristics and hydrophobicity, respectively. For metal ions, the variable that best modeled the toxicity data obtained with the two T. pyriformis tests was the softness index [sigma(p), i.e., (coordinate bond energy of the metal fluoride--coordinate bond energy of the metal iodide)/(coordinate bond energy of the metal fluoride)]. No correlation was found with the Microtox test. For organic compounds, a significant correlation was observed between the hydrophobicity coefficient and the toxicity data. This correlation is closer with the two tests using Tetrahymena. PMID- 11440485 TI - Particulate air pollution, C-reactive protein, and cardiac risk. PMID- 11440486 TI - Coronary angiogenesis: dipyridamole is back on the stage? PMID- 11440487 TI - Risk of sudden death after discharge following myocardial infarction. PMID- 11440488 TI - Cell adhesion molecules and inflammation in acute coronary syndromes: markers and emerging risk factors. PMID- 11440489 TI - Cardiac surgery in octogenarians: who, when and how? PMID- 11440490 TI - A classification of atrial flutter and regular atrial tachycardia according to electrophysiological mechanisms and anatomical bases; a Statement from a Joint Expert Group from The Working Group of Arrhythmias of the European Society of Cardiology and the North American Society of Pacing and Electrophysiology. PMID- 11440491 TI - Multivessel coronary artery disease: current revascularization strategies. PMID- 11440492 TI - Particulate air pollution is associated with an acute phase response in men; results from the MONICA-Augsburg Study. AB - AIMS: Episodes of increased air pollution are associated with increases in hospital admissions for cardiovascular disease. Even modest acute phase responses are associated with increased risk of coronary heart disease. The study investigates whether induction of an acute phase response by exposure to air pollution may contribute to cardiovascular pathology. METHODS AND RESULTS: A prospective cohort study based on a survey in 1984/85 with a 3-year follow-up was conducted in 631 randomly selected men aged 45 to 64 years free of cardiovascular disease at entry 1984/85. Serum C-reactive protein concentrations were determined by a high sensitivity immunoradiometric assay. C-reactive protein concentration was increased in association with the 1985 air pollution episode. In multivariate analyses, elevated concentrations were independently associated with concentrations of total suspended particles and the sulphur dioxide episode. At ambient concentrations of pollution, as noted during the 1985 air pollution episode, the odds of observing C-reactive protein concentrations above 5.7 mg. l( 1)(>90th percentile) tripled, and increases of 26 microg. m(-3)total suspended particles (mean of 5 days) raised the odds of C-reactive protein levels 50% above the 90th percentile. CONCLUSIONS: Exposure to current levels of particulate matter in the atmosphere elicits an acute phase response in randomly selected healthy middle-aged men, which may contribute to the increased cardiovascular risk caused by air pollution. PMID- 11440493 TI - Effects of dipyridamole on coronary collateralization and myocardial perfusion in patients with ischaemic cardiomyopathy. AB - AIMS: There is evidence that oral dipyridamole, a nucleoside uptake blocker that increases myocardial adenosine levels, lessens myocardial ischaemia by inducing coronary collateral growth in animal models of ischaemic heart disease. However, whether dipyridamole can exert a similar effect in humans with coronary artery disease is controversial. METHODS AND RESULTS: We studied 30 male patients (mean age 55+/-9 years) with coronary artery disease and left ventricular systolic dysfunction (ejection fraction >40%). Patients were randomized into three matched groups. Group A patients (n=10) received dipyridamole alone at a dose of 75 mg t.i.d. orally for 8 weeks. Group B patients (n=10) underwent exercise training at 60% of peak .VO(2)three times a week for 8 weeks, and received dipyridamole. Group C patients (n=10) had neither exercise testing nor dipyridamole. On study entry and after 8 weeks all patients underwent an exercise test with gas exchange analysis, dobutamine stress echocardiography, 201-thallium planar myocardial scintigraphy, and coronary angiography. Peak .VO(2)increased significantly only in trained patients. Thallium uptake of the collateral-dependent myocardium, coronary collateral score and wall thickening score increased significantly only in groups receiving dipyridamole, the greatest improvement being in group B patients. Plasma adenosine levels were also the highest in group B (P<0.001 vs A and C). Correlations were found between changes in adenosine levels and increases of both thallium uptake (r=-0.70;P=0.001) and collateralization (r=0.72;P=0.001). CONCLUSION: Exercise training potentiates the effects of dipyridamole on coronary collateralization and myocardial perfusion in humans with ischaemic cardiomyopathy. PMID- 11440494 TI - Determinants of sudden death after discharge from hospital for myocardial infarction in the thrombolytic era. AB - AIMS: The purpose of this study was to assess risk factors for sudden death after discharge from hospital for myocardial infarction in an era in which 50% of patients receive thrombolytic drugs. METHODS AND RESULTS: We prospectively studied 708 consecutive survivors of myocardial infarction admitted to hospitals, which had registered their clinical, functional, and electrical parameters. A total of 83 patients died in the first 2 years (12%) after discharge. Sudden death was only observed in 12 patients. In multivariate analysis only NYHA class >I, and a filtered QRS duration > or =110 ms were important predictive variables for sudden death. A pre-defined high-risk group of 25 patients had no sudden death. When the strongest predictive variables in univariate analysis were combined to increase the positive predictive value for sudden death, we only achieved a maximal value of 27%. CONCLUSIONS: In an unselected infarction population, the risk for sudden death is low in the first 2 years. Therefore, prediction and prophylactic intervention, such as defibrillator therapy become difficult. The event is related to cardiac dysfunction on admission, and with abnormalities in the filtered electrocardiogram. PMID- 11440495 TI - Early increase in levels of soluble inter-cellular adhesion molecule-1 (sICAM-1); potential risk factor for the acute coronary syndromes. AB - BACKGROUND: Studies have shown disparate results in relation to the role of plasma concentrations of cell adhesion molecules in atherosclerosis. Moreover, the differentiation of primary vs secondary alterations of these markers, in response to myocardial injury, has not been clear. We measured specific soluble cell adhesion molecules and inflammatory markers in men admitted acutely with chest pain and compared them to healthy controls. METHODS AND RESULTS: We prospectively studied men (total n=241), admitted acutely with chest pain (7.4+/ 9.4 h, 71% within 10 h), unstable angina (n=67), acute myocardial infarction (n=47) and chest pain without ischaemic heart disease (n=45) and compared them with a stratified sample of randomly selected healthy controls (n=82). Soluble intercellular adhesion molecule (sICAM-1), endothelial selectin, vascular cell adhesion molecule, interleukin-6 and C-reactive protein were measured by ELISA and P-selectin expression by flow cytometry. Multiple regression analysis was used to control for the impact of classical risk factors. At baseline ICAM-1, interleukin-6 and C-reactive protein were significantly elevated in patient groups whereas no difference in vascular cell adhesion molecule or endothelial selectin was found. At 3 month follow-up, ICAM-1 level was unchanged in ischaemic heart disease patients. In all groups C-reactive protein and interleukin-6 levels were lower at review. ICAM-1 levels at follow-up were higher in ischaemic heart disease groups (but not in chest pain without ischaemic heart disease) relative to controls and remained so only in the unstable angina group following regression. sICAM-1, interleukin-6 and C-reactive protein strongly correlated with smoking. In the acute phase, ICAM-1 was confounded by smoking following regression and C-reactive protein and interleukin-6 remained significant in both ischaemic heart disease groups after multiple regression. There was no relationship to events which occurred in 23% of ischaemic heart disease patients (further acute myocardial infarction 5.3%, sudden cardiac death 0.9% or recurrent angina 16.7%). CONCLUSION: We found an inflammatory response with higher sICAM-1, interleukin-6 and C-reactive protein in patients presenting soon after developing an acute coronary syndrome. As sICAM-1 was not affected by the acute event this plasma marker may be an important risk factor for the development of the acute coronary syndrome, particularly unstable angina. PMID- 11440496 TI - Cardiac surgery in octogenarians; peri-operative outcome and long-term results. AB - AIMS: Because the elderly are increasingly referred for operation, we reviewed the results of cardiac surgery in patients of 80 years or older. METHODS AND RESULTS: Records of 182 consecutive octogenarians who had had cardiac operations between 1992 and 1998 were reviewed. Follow-up was 100% complete. Seventy patients had coronary grafting (CABG), 70 aortic valve replacement, 30 aortic valve replacement+CABG, and 12 mitral valve repair/replacement. Rates of hospital death, stroke, and prolonged stay (>14 days) were as follows: CABG: 7 (10%), 2 (2.8%) and 41 (58%); aortic valve replacement: 6 (8.5%), 2 (2.8%) and 32 (45.7%); aortic valve replacement+CABG: 8 (26.5%), 1 (3.8%) and 14 (46.6%); mitral valve repair/replacement: 3 (25%), 1 (8.3%) and 5 (41.6%). Multivariate predictors (P<0.05) of hospital death were New York Heart Association functional class, urgent procedure, prolonged cardiopulmonary bypass time, and, after aortic valve replacement, previous percutaneous aortic valvuloplasty. Ascending aortic atheromatous disease was predictive of stroke, while pre-operative myocardial infarction was predictive of prolonged hospital stay. Actuarial 5-year survival was as follows: CABG, 65.8+/-8.8%; aortic valve replacement, 63.6+/-7.1%; aortic valve replacement+CABG, 62.4+/-6.8%; mitral valve repair/replacement, 57.1+/ 5.6%; and total, 63.0+/-5.6%. Multivariate predictors of late death were pre operative myocardial infarction, and urgent procedure. Ninety percent of long term survivors were in New York Heart Association class I or II, and 87% believed having a heart operation after age 80 years was a good choice. CONCLUSION: Cardiac operations are successful in most octogenarians with increased hospital mortality, and longer hospital stay. Long-term survival and quality of life are good. PMID- 11440498 TI - Peripheral mobilisations with movement. AB - SUMMARY. The use of mobilisations with movement (MWM) for peripheral joints has been developed by Mulligan. A mobilisation is applied parallel or at right angles to the restricted joint movement. If the applied mobilisation achieves immediate improvement in the functional movement and abolishes the pain the treatment involves sustaining the mobilisation while the patient performs the active movement repetitively. On reassessment of the joint function, the movement should remain improved without the mobilisation. Theories as to why these techniques provide rapid improvement in pain-free range are proposed, and the general principles of examination and treatment are outlined. Specific clinical examples demonstrate how MWM can be used in isolation or integrated with other manual approaches to improve the quality of joint intraarticular gliding, neurodynamics and the facilitation of correct muscle recruitment. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440499 TI - Pathogenesis of upper cervical instability. AB - SUMMARY. The purpose of this article is to review the pathogenesis of upper cervical instability. Instability can arise from inflammatory, congenital and traumatic causes. The commonest causes of atlanto-axial dislocation are rheumatoid arthritis and Down's syndrome. The review revealed much less information about the relatively minor instabilities that are probably responsible for a number of chronic complaints of the type seen by manual therapists. The potential involvement of passive (ligamentous), active (musculotendinous) and neural control subsystems in maintaining the stability of the spine may go some way to explaining the spectrum of conditions included under the term clinical instability. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440500 TI - The muscular arch of the axilla revisited: its possible role in the thoracic outlet and shoulder instability syndromes. AB - SUMMARY. The thoracic outlet syndrome and shoulder instability provoke well known and partly similar sensations, often described by the patient as "my arm feels deadellipsisI have no strength in my armellipsisI feel pins and needles in my armellipsis". In axillary surgery, there are sometimes reports of a fibromuscular band or string crossing the axilla. In the dissection room these bands are well known. Their reported incidence being about 10% at post mortem. Generally they are variations of the latissimus dorsi or the pectoralis major muscles, the most common form being known as the axillary arch or the arch of Langer. Simulations of abduction in combination with external rotation on dissected cadaveric material suggest a possible neurovascular compression at the transition from the axilla to the upper end of the brachial neurovascular bundle. Dissection confirms that the band is innervated by branches from the ansa pectoralis with a blood supply from the lateral thoracic artery. Following a case study, where the axillary arch negatively influenced the patients occupational skills, an extensive echographic study of 1321 subjects (1179 male, 142 female) was undertaken. A total of 188 arches was detected in 112 subjects (8.50%), mostly occurring bilaterally (incidence 5.80%). The incidence of the arch of Langer suggests that it should be considered in the differential diagnoses of thoracic outlet and shoulder instability syndromes. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440502 TI - Digital videofluoroscopy. PMID- 11440501 TI - Measurement of shoulder joint kinaesthesia. AB - SUMMARY. Loss of kinaesthesia has been identified as a factor in recurrent or persistent problems following glenohumeral dislocation. However, accurate measurement of shoulder kinaesthesia remains a difficult task in the clinical setting. This study investigated the use of the Kincom dynamometer for measuring the ability of a subject to reproduce a shoulder joint angle with a passive and active test. The tests were applied to two subject groups, (normal and patient) and the results were compared with those of a standard clinical test. The results of the Kincom tests demonstrate a statistically significant difference between the normal and patient groups (F((1,140)) = 53.0; P < 0.001). The difference between the means for the two groups was 1.2 degrees which indicates the precision of shoulder kinaesthetic sense. All subjects were more accurate with the active test compared to the passive test (F((1,140)) = 36.5; P < 0.001). Reliability of the Kincom tests was evaluated over 3 days and a significant difference was found between day one and day three for both groups (F((2,140)) = 3.5; P < 0.05) suggesting a learning effect had occurred. There was no significant difference between the Kincom active test and the clinical test (F((1,28)) = 1.86; P = 0.18); however, the difference between the Kincom passive test and the clinical test was significant (F((1,28)) = 10.74; P < 0.01). The clinical test did not demonstrate a statistically significant difference between the two subject groups (t(28 = 1); P = 0.05). Copyright 1996 Harcourt Publishers Ltd. PMID- 11440503 TI - Vertebral artery testing - the Australian Physiotherapy Association Protocol after 6 years. PMID- 11440504 TI - Positive Upper Limb Tension Test in a Case of Surgically Proven Neuropathy: Analysis and Validity. PMID- 11440505 TI - The management of low back pain in pregnancy. AB - SUMMARY. Low back pain is a common condition seen in pregnancy. The treatment of low back pain in pregnant patients is essentially different from the treatment of the non-pregnant patient. These differences arise when it comes to understanding the reasons why pregnant patients get back pain and the differential diagnostics relating to obstetrics. The techniques are different as the patient cannot be treated prone, and manual techniques are better than mechanical ones because the palpatory response to the changing tissues is very important in this group of patients. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440506 TI - Tendons in health and disease. AB - SUMMARY. This article reviews the structure, functions and mechanical properties of normal tendons and surveys the range and causes of tendon diseases. Specialized regions of tendons are considered, including the myotendinous junction, enthesis and regions where tendons rub against bone. Particular emphasis is placed on recent literature and on aspects of tendons that are relevant to manual therapists. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440507 TI - An investigation of neural tissue involvement in ankle inversion sprains. AB - SUMMARY. The slump test was used to investigate the effect of ankle inversion sprain on neurodynamic function in 18 subjects. The sensory response and restriction in range of knee extension in the slump test were assessed. Three different foot and ankle positions were used-neutral, dorsiflexion (DF) and plantarflexion with inversion (PFI). Results indicated that although there was a significantly greater lack of knee extension on the injured side compared to the non-injured side in all three foot and ankle positions, knee extension was most restricted in the PFI position, which is considered to bias the common peroneal tract. The slump test in PFI produced symptoms in the lateral aspect of the lower leg and ankle extending slightly anteriorly and posteriorly and into the dorsum of the foot. This distribution corresponds to that of the superficial peroneal nerve. The slump test in neutral and DF produced areas of response similar to each other, which included the posterior aspect of the knee, thigh and calf. The release of cervical flexion resulted in a significant reduction in symptoms in each test. These results may indicate altered neurodynamic function following ankle inversion sprain, and have implications for assessment and treatment of subjects with ankle sprain. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440508 TI - To compare the effects of different rates of application of a cervical mobilisation technique on sympathetic outflow to the upper limb in normal subjects. AB - SUMMARY. The aim of this study was to compare the effects of different rates of application of a commonly used physiotherapeutic spinal manipulative therapy technique on sympathetic function in normal pain-free volunteers. A randomized, repeated measures, double blind, controlled study design was used to investigate the effects of two different rates of a C5 grade III central postero-anterior mobilization technique on skin conductance (SC) and skin temperature (ST) in the distal C6 dermatome of asymptomatic subjects. Sixteen asymptomatic male volunteers participated in the study. Application of a C5 central postero anterior grade III mobilization at the rate of 2Hz produced significantly greater increases in SC values than that at the rate of 0.5 Hz and control.The results of this study suggest that mobilization with the rate which is commonly used clinically causes a greater increase in sympathetic efferent activity in the upper limb of normal pain-free volunteers than a slower rate. These results may also provide a basis for further investigation into the physiological effects of different rates of mobilization and in particular exploration of the relationship between changes in sympathetic function following mobilization and manipulation induced analgesia. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440509 TI - Instrumented measurement of spinal stiffness. AB - SUMMARY. The characteristics of a safe, portable and accurate spinal stiffness testing device are described in this report. Because judgements of spinal stiffness made using manual tests have been shown to be unreliable, several mechanical devices have been designed to measure posteroanterior (PA) stiffness. One major limitation of these devices has been the inability to transport them to locations where subjects with spinal pain could be easily accessed and measured. This paper reports on the design and fabrication of a new, portable, mechanical device for measuring PA stiffness in the lumbar spine. Information regarding the safety features of the device, and contraindications for its use are provided. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440510 TI - Expanding the scope of physiotherapy: clinical physiotherapy specialists in consultants' clinics. PMID- 11440511 TI - The Role of Transversus Abdominus in Post Partum Correction of Gross Divarication Recti. PMID- 11440512 TI - Rotational instability of the mid-thoracic spine: assessment and management. AB - SUMMARY. Recent research has enhanced the understanding of instability of the spine. The principles of this research have been incorporated into the evaluation and treatment of the unstable thorax. Rotational instability of the midthorax is commonly seen following trauma to the chest. Specific mobility and stability tests have been developed to detect this instability. The tests are derived from a biomechanical model of evaluation. Treatment is based on sound stabilization principles and although the segment will remain unstable on passive testing, the patient can be trained to control the biomechanics of the thorax and return to a high level of function. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440513 TI - Biomedical considerations and clinical patterns related to disorders of the glenoid labrum in the predominantly stable glenohumeral joint. AB - SUMMARY. The roles of the glenoid labrum and long head of biceps are reviewed together with their significance in stability of the glenohumeral joint. Clinical presentations related to disorders of the glenoid labrum and long head of biceps not associated with frank instability but commonly responsible for dysfunction in the athletic shoulder, are reviewed from the perspective of pertinent anatomical, biomechanical and kinematic knowledge. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440514 TI - Diagnostic tests for temporomandibular disorders: an assessment of the methodologic quality of research reviews. AB - SUMMARY. The objective of this study was to assess review articles of temporomandibular disorder (TMD) diagnostic tests for methodological quality and to establish the need for a systematic reviewing process. Computerized bibliographic database searches from 1983 to December 1996, information requests from authors, and review of reference lists of all retrieved reports were used to identify relevant reviews. After applying selection criteria, two investigators independently reviewed the articles. Each review article was evaluated independently for methodological quality. Agreement between reviewers was measured using quadratic weighted kappa.Forty-five of 53 review articles met the selection criteria. No review described the search methods, selection criteria, demonstrably avoided selection bias or pooled data of primary studies. Three of 45 articles assessed the internal validity and reliability of the studies included in their review. Major to extensive methodological flaws were noted across all reviews.This critique of methodologic quality in TMD diagnostic review articles supports the need for a systematic reviewing process. Clinicians turning to diagnostic reviews to aid them in synthesizing a large body of literature may wish to apply the described methodologic criteria to separate 'valid' from 'invalid' reviews. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440515 TI - Pressure algometry in manual therapy. AB - SUMMARY. The pressure pain threshold (PPT), i.e. the least stimulus intensity at which a subject perceives pain, was studied in 30 patients with chronic unilateral pain in the shoulder and arm region.Fourteen trigger points were investigated on both sides of the body using pressure algometry. Eight paravertebral points and six points in the shoulder and arm region were evaluated. Each location was examined twice. The patients were tested by one examiner.The intraobserver reliability of PPT measurements was considered to be good. The Intraclass Correlation Coefficients of reliability (ICC), based on two repeated PPT measurements varied between 0.64 and 0.96. The painful side of the body was found to be more sensitive than the non-painful side, although there was not always a significant difference at the 0.05 level. The PPT was found to be higher in males than in females (P < 0.05). Females demonstrated lower PPTs than males at each trigger point. It was established that pressure tenderness varies over individual trigger points. Significant regional differences in PPT values were observed (P < 0.05). PPT values decreased in a cranial direction in the spine and in a caudal direction in the upper limb.Although the authors expected to find segmentally reduced PPT values on the painful side of the body, a generalized reduction of PPT values was present at all peripheral and spinal segmental sites. Some correlations between segmentally related trigger points were found by factor analysis. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440516 TI - The effect of training with feedback on physiotherapy students' ability to judge lumbar stiffness. AB - SUMMARY. Previous studies have demonstrated that manual judgments of lumbar posteroanterior (PA) stiffness show poor reliability. One explanation for this poor reliability may be that the method of training students using feedback provided by physiotherapy tutors is ineffective. The aim of the current study was to investigate whether immediate quantitative feedback, provided from a highly reliable mechanical device, could improve physiotherapy students' ability to judge lumbar PA stiffness. Four physiotherapy student raters assessed 75 stiffness stimuli (provided by the lumbar spines of asymptomatic volunteers) during pre-test, training and post test sessions held over a 3 week period. During the training sessions raters were provided with accurate and immediate feedback regarding each judgment of PA stiffness at the L3 vertebral level of asymptomatic lumbar spines. No significant difference in mean absolute error between the pre and post tests was found (P = 0.31). Provision of information about the true PA stiffness of each lumbar spine judged, therefore, did not improve the accuracy of physiotherapy students' judgments of lumbar stiffness. Copyright 1996 Harcourt Publishers Ltd. PMID- 11440518 TI - LATERAL LIGAMENT STRAINS OF THE ANKLE, DO THEY EXIST? PMID- 11440517 TI - Can manual therapy benefit from the Cochrane Collaboration? PMID- 11440519 TI - Sacroiliac dysfunction in dancers with low back pain. AB - SUMMARY. Low back pain is a common occurrence in dancers. Studies have shown its prevalence to be around 12% of all dance type of injuries. It is commonly thought by health professionals who specialize in dance medicine that sacroiliac (SI) dysfunction is one of the more common causes of low back pain in dancers. The aetiology of SI dysfunction in dancers is related to both the biomechanics of the SI joint and the physiological demands placed on the SI joint from the dynamics of dance. Injury to the SI joint can be due to a combination of a single traumatic incident, from overuse factors involving repetitive microtrauma or from emotional stress. Clinical manifestations could be pain in the back, buttock, hip and leg, and limitation of movement specific to dance. Diagnosis is based upon the deviation from normal of both the static and kinetic functions of the low back and pelvis in its relationship to the biomechanics of dance. Treatment is aimed at relieving pain, restoring the function of the SI joint and returning the dancer to full function. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440520 TI - The effects of joint damage on muscle function, proprioception and rehabilitation. AB - SUMMARY. The paper summarizes a series of studies investigating reduced quadriceps femoris activation and its effect on rehabilitation of patients with traumatic and arthritic knee damage. It incorporates findings from related research fields and speculates on the physiological causes of reduced voluntary activation following joint damage and its clinical implications. Arthrogenic reduction in voluntary activation is the inability to maximally activate muscles acting across damaged joints. This phenomenon may cause muscle weakness by preventing complete activation of the muscle and if prolonged may result in muscle fibre atrophy, and may possibly impede rehabilitation. In patients with anterior cruciate ligament (ACL) deficient knees the reduction in quadriceps femoris activation was related to the amount of joint damage sustained and caused some quadriceps femoris weakness. Patients with isolated ACL ruptures had a small reduction of quadriceps femoris activation that did not impede conservative rehabilitation. However, patients with ACL ruptures and associated joint damage had a large reduction in activation and responded poorly to rehabilitation. Operative stabilization of ACL-deficient limbs may increase activation and improve efficacy of rehabilitation. Quadriceps femoris activation was also reduced in patients with mild knee osteoarthritis, who responded well to rehabilitation. Following joint damage, abnormal articular afferent information may decrease alpha-motoneurone excitability, reducing voluntary quadriceps femoris activation. If joint damage is extensive the resulting large reduction in activation may prevent the threshold for stimulation of muscle hypertrophy from being reached, which impedes rehabilitation. Abnormal articular afferent information may also decrease gamma-motoneurone excitability causing proprioceptive deficits. Rehabilitation which increases alpha-motoneurone excitability may also increase gamma-motoneurone excitability, improving proprioception. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440521 TI - The influence of an anteroposterior accessory glide of the glenohumeral joint on measures of peripheral sympathetic nervous system function in the upper limb. AB - SUMMARY. The present study was undertaken to determine whether a sympathoexcitatory effect would result from a manipulative therapy technique applied to a peripheral joint. The technique selected was a Grade III posteroanterior accessory glide of the right glenohumeral joint with the arm positioned in glenohumeral abduction and internal rotation. Previously, spinal manipulative therapy techniques have been shown to produce such an effect. Nineteen normal subjects participated in the experiment. Treatment, placebo and control conditions were compared. Skin conductance and skin temperature of the distal finger tips were used as indicators of peripheral sympathetic activity. Results demonstrated that the treatment condition caused a significantly greater increase in skin conductance than the placebo or control conditions. The response was seen in both hands indicating a generalised sympathetic activation. The treatment condition also resulted in a significant reduction in skin temperature in the treated limb. The importance of this sympathoexcitatory effect in relation to potential neurophysiological mechanisms underlying manipulation induced analgesia is discussed. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440523 TI - Quality in practice-a guide to developments in the UK's health service. PMID- 11440522 TI - Left-right asymmetries and other common anatomical variants of the first cervical vertebra. AB - SUMMARY. Left-right asymmetries were systematically and independently observed twice in 82 macerated atlases by three investigators using a check-list and the instructions of an examination protocol. A grid behind the vertebra served as a visual aid and was inserted in an inclinometer box used for measuring zygapophyseal joint facet inclination. Only those asymmetries observed once by each of the investigators were listed. The posterior arch showed the highest number of asymmetries and many unequal grooves for the vertebral artery were observed. The lateral compartment frequently revealed asymmetries of transverse processes and transverse foramina. These results are indicative of variants of the bony canal for passage of the vertebral artery and asymmetric lever arms for muscles, inserted at C1. A third zone could be defined at the level of the joint facets. Tropism of the superior joint facets was caused by differences in shape, size, surface area, curvature, inclination relative to the sagittal plane, implantation on the lateral mass, unilateral or asymmetric subdivision. Left right differences of inferior joint facets were often related to degenerative enlargement. Osteophytes also caused asymmetry of the facet for the odontoid process. An important number of combined asymmetries reveals questions about their functional consequences. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440524 TI - Rehabilitation of Kinaesthesia in the Shoulder. PMID- 11440525 TI - Dynamic stability of the scapula. AB - SUMMARY. The ability to position and control movements of the scapula is essential for optimal upper limb function. The inability to achieve this stable base frequently accompanies the development of shoulder and upper limb pain and pathology. Unlike other joints the bony, capsular and ligamentous constraints are minimal at the scapulothoracic 'joint' so stability is dependant on active control. Clinically, it is noted that patients presenting with shoulder and arm symptoms demonstrate poor dynamic scapula control. Scapula setting is an exercise taught by physiotherapists to correct movement dysfunction associated with abnormal scapula positioning and dynamic control. Addressing the dynamic stabilization of the scapula is an essential part of the management of neuromusculoskeletal dysfunction of the shoulder girdle and an appropriate rehabilitation programme is necessary if this issue is to be addressed. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440526 TI - Thoracic spine: anatomical and biomechanical considerations for manual therapy. AB - SUMMARY. Interactions between thoracic spine posture and mobility are believed to play a role in the development of spinal pain syndromes. Accordingly, appropriate and effective manual therapy management is dependent on a sound knowledge of the anatomy and biomechanics of this region of the vertebral column. This paper reviews the primary anatomical influences on thoracic posture, mobility and stability. The consequences of anatomical variation and degenerative change, their effect on thoracic kyphosis, and the range and patterns of thoracic movement, are examined. The implications for assessment and management of thoracic spine dysfunction are discussed together with some considerations for the application of manual therapy treatment. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440527 TI - Upper trapezius muscle activity during the brachial plexus tension test in asymptomatic subjects. AB - SUMMARY. The brachial plexus tension test (BPTT) is used clinically to test the dynamics of the neural tissues of the upper quadrant. The upper trapezius muscle and the nerves of the brachial plexus share common anatomical locations and are jointly affected by BPTT movements. This study investigated the relationship between the BPTT, upper trapezius muscle activity and range of neural tissue extensibility in asymptomatic subjects. Normal male subjects with greater and lesser neural tissue extensibility were tested. Results revealed that those with lesser neural extensibility exhibited significantly greater upper trapezius muscle activity during discrete BPTT stages. There was no difference between groups in the levels of pain perceived with the test. These results suggest that asymptomatic neural tissues are protected from stretch by muscle activity not solely mediated by pain but also possibly mediated by stretch receptors in neural structures. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440528 TI - Reliability of a discrimination measure for judgements of non-biological stiffness. AB - SUMMARY. This study determined the just noticeable difference, or discrimination threshold, for manual stiffness perception and the test-retest reliability of this measure. Elastic, non-biological stiffness stimuli within the range found in human spines were generated by a device incorporating metal springs. The method of constant stimuli was used to estimate the just discriminable change in stiffness and results were expressed as a percentage of the base stiffness, or Weber fraction. A total of 25 physiotherapists and lay people participated on two separate occasions. Reliability of repeated measurements was evaluated by intraclass correlation coefficients, percentage of agreement scores and the standard error of the measurement. All three reliability coefficients showed high/good reliability for the repeated measurements. The mean Weber fraction was 7.7%, implying high sensitivity to differences in non-biological stiffness. The stability of the Weber fraction means that this index can be used as a measure of a subject's sensitivity to stiffness stimuli. Consequently, training programmes and various manual techniques designed to enhance therapists' stiffness discrimination can be evaluated. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440529 TI - Overuse syndromes of the upper limb and the upper limb tension test: a comparison between patients, asymptomatic keyboard workers and asymptomatic non-keyboard workers. AB - SUMMARY. Overuse syndromes of the upper limb (OSUL), also known as repetitive strain injury (RSI) remains a contentious issue that affects a large number of keyboard workers. This research used the results of a modified upper limb tension test (ULTT) performed on three independent groups to look for any similarities or discrepancies between them. The subjects were all female aged between 25-40 years, with groups consisting of 12 OSUL patients, 20 asymptomatic keyboard users and 19 asymptomatic non-keyboard users. The ULTT was positive in 100% of the patient group supporting several hypotheses that state that the pathology of OSUL is neurogenic in origin. The results of the two asymptomatic control groups were predicted to be the same, however this was not found to be the case. Unrelated t tests were used to analyse the data showing that the keyboard users had a significantly positive ULTT compared to non-keyboard users, which were viewed as a normal control group. This suggests that a subclinical pathological state exists in the keyboard user group. Various subclinical physiological changes caused by prolonged sitting and keyboard use are suggested as possible causes for these results. This research highlights the need for intervention in an office environment to prevent further cases of OSUL from developing. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440530 TI - Central facilitation and remote effects: treating both ends of the system. AB - SUMMARY. This article reports a series of three case studies where the author carried out a spinal mobilization treatment concurrent with the patient's attempt to contract a chronically weakened muscle or muscle group. All of the cases describe enhancement of the contraction of the posterior crural muscles post trauma. A brief discussion of the results and potential mechanisms by which the results may have been achieved is presented. This involves reference to proprioceptive neuromuscular facilitation (PNF), muscle energy techniques and to spinal mobilizations with limb movements. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440531 TI - Management of cervical headache. AB - SUMMARY. The success of physical therapies in the management of headache relies in the first instance on an accurate differential diagnosis of a cervical musculoskeletal origin to the headache. Examination should identify a symptomatic pattern of headache characteristic of neck dysfunction and these symptoms must be associated with relevant physical impairments in the cervical articular and muscle systems. Dysfunction in the upper three cervical joints, poor activation levels and endurance capacity of the deep and postural supporting muscles of the neck, shoulder girdle region and deficits in kinesthesia have been identified in the cervical headache patient. Treatment needs to be precise and comprehensive to address each aspect of this interrelated dysfunction if long-term success of treatment is to be achieved. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440532 TI - Retrodiscal tissue of the temporomandibular joint: clinical anatomy and its role in diagnosis and treatment of arthropathies. AB - SUMMARY. Physiotherapists are working increasingly with the dental profession to treat patients with craniomandibular disorders. Temporomandibular arthropathies are some of the more difficult conditions to treat. The retrodiscal tissues of the articular complex can be injured and be a source of pain or they may contribute to the development of arthropathies. This review will consider the clinical anatomy, the kinematics and the pathology of the retrodiscal tissues in the bilaminar zone of the temporomandibular joint. Their role in the etiology of temporomandibular arthropathies is presented and proposals for manual testing and treatment are made. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440533 TI - Determination of manipulative physiotherapy treatment outcome in headache patients. AB - SUMMARY. Although physiotherapists are frequently involved in the treatment of headache when cervical spine dysfunction is thought to be a cause or contributing factor there does not appear to be a consistent definition of treatment success. This study analysed treatment outcome in a population of 112 headache patients presenting for manipulative physiotherapy. Two months after the initial consultation, statistically significant improvements were observed in mean scores for each of headache frequency (P < 0.001), duration (P < 0.05) and intensity (P < 0.001). When a combination of patient estimate of treatment effect and a headache index incorporating scores for frequency, intensity and duration was used to classify treatment outcome, 51 of the 91 subjects analysed at follow-up were deemed to have had a positive response to treatment. This method of classification of treatment outcome appeared to be sensitive to cases where the patient's headaches were improved by factors other than the physiotherapy treatment. However the classification used was less sensitive when the patient's headaches were aggravated by other factors or where there was not consistent improvement or deterioration in the measures of headache frequency, intensity and duration. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440534 TI - Validity and reliability of measures obtained from the OSI CA-6000 Spine Motion Analyzer for lumbar spinal motion. AB - SUMMARY. The aims of this investigation were firstly to determine the level of agreement between angular measures of lumbar spinal motion recorded by the OSI CA 6000 Spine Motion Analyzer (OSI SMA) and measures obtained from X-rays of subjects without lumbar symptoms, and secondly to determine the intra-rater reliability of measures of lumbar spinal range of motion using the OSI SMA on individuals with lumbar symptoms. To fulfill the first aim 13 asymptomatic subjects (mean age = 28.6 years; SD = 7.1) performed the motions of lumbar flexion, extension, side bending to the right, and side bending to the left twice while wearing the OSI SMA, and once for the exposure of the X-ray. The difference between the measures obtained by the two methods was plotted against the average of those measures for each subject to illustrate the level of agreement of the two methods. Differences between measures that were within one standard deviation of the mean were as follows: for flexion, 7 out of 10 showed less than 8 degrees difference; for extension, 7 out of 10 showed less than 7 degrees difference; for side bending to the right, 11 out of 13 showed less than 6 degrees difference; and for side bending to the left, 10 out of 13 showed less than 5 degrees difference. For asymptomatic subjects, measures obtained from the OSI SMA appear to provide good agreement with measures of lumbar spinal motion in the sagittal and frontal planes as determined by X-rays. To fulfil the second aim of the investigation thirty subjects with lumbar symptoms (mean age = 35.9 years; SD = 14.2) performed four trials of lumbar motion to their maximum for flexion, extension, side bending to the right and left, and rotation to the right and left. The OSI SMA hardware was then completely removed and replaced by the same examiner, and the trials were repeated. ICC values were 0.903 or higher for all motions. These results suggest that active range of motion measures obtained with the OSI SMA in subjects with lumbar symptoms are consistent over repeated trials. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440535 TI - Influence of a cervical mobilization technique on respiratory and cardiovascular function. AB - SUMMARY. Spinal manipulative therapy techniques are frequently applied by physiotherapists to relieve pain of musculo-skeletal origin and to improve the quality of joint movement in a variety of musculo-skeletal conditions. However, there has been little research into the physiological effects of these techniques, or the mechanisms responsible for these effects. The aim of this study was to establish whether a grade III posteroanterior mobilization technique applied centrally to the cervical spine would affect respiratory and cardiovascular indicators of sympathetic nervous system function in pain-free, normal volunteers. A significant increase in respiratory rate, heart rate, systolic and diastolic blood pressure occurred during application of the technique to C5/6, when compared to the control and placebo conditions. There was little change in any of the measured variables during the placebo condition. This study provides objective evidence that application of this mobilization technique elicits changes in sympathetic nervous system activity distinct from placebo in pain-free individuals. These results provide a basis for further research into the physiological effects of manipulative procedures, and in particular, exploration of the mechanisms responsible for analgesia produced by this method. Copyright 1997 Harcourt Publishers Ltd. PMID- 11440536 TI - Focal dystonia in a musician. PMID- 11440537 TI - Immunocytochemical localization of a caveolin-1 isoform in human term extra embryonic membranes using confocal laser scanning microscopy: implications for the complexity of the materno-fetal junction. AB - This immunochemical, immunocytochemical, histological and ultrastructural study demonstrates the presence of caveolin 1 in a number of locations in term human extra-embryonic membranes. Strong expression was observed in fetal blood vessel endothelial cells of chorionic villi (cv) and in cv, amniotic and chorionic plate mesenchymal cells, but weak expression was characteristic of trophoblast. Expression in the amniotic epithelium indicated a stronger association with apical as opposed to baso-lateral membranes. Strong immunoreactivity in the thin lining layer of the maternal blood space of the basal plate was a surprising finding. Previously defined as trophoblast, we argue that this is at least partly endothelium based on this new histological, ultrastructural and immunocytochemical data. PMID- 11440538 TI - The Human Achaete Scute Homolog 2 gene contains two promotors, generating overlapping transcripts and encoding two proteins with different nuclear localization. AB - Placental development involves control by the basic helix-loop-helix transcription factor Mash2. Transcript analysis of the Human Achaete Scute Homolog 2 (HASH2) mRNA revealed the presence of two overlapping transcripts in first trimester placentae. The two transcripts (2.6 and 1.5 kb) are generated by two promotors which are separated by 1.1 kb, generating transcripts 1 and 2, respectively. Surprisingly, in transcript 1 which shows a broad expression, a second potential coding region, tentatively called Human Achaete Scute Associated Protein (HASAP) was present. Transcript 2 contains the HASH2 encoding region only. Analysis of protein expression from both transcripts by transfection studies with eGFP fusion proteins, revealed that both coding regions are translated from their endogenous translation initiation site and showed that both proteins are transported to the nucleus. HASH2 is distributed throughout the nucleus but the HASAP protein is transported into nuclear compartments, the nucleoli. In addition, the HASAP protein lacks the bHLH domain and bears no homology to known proteins. Moreover, allele-specific RT-PCR showed the human gene not to be subject to imprinting, possibly reflecting the biallelic expression of one of both transcripts. Our data indicate a species-specific difference between mouse and human expression of the Achaete Scute Homolog 2 and suggests a dual function of the human homologue. PMID- 11440539 TI - Hypoxia attenuates PGE(2)but increases prostacyclin and thromboxane production in human term villous trophoblast. AB - Prostanoids have been proposed to play a major role in the regulation of uteroplacental blood flow. We examined the effect of hypoxia on the production of prostaglandin E(2)(PGE(2)) thromboxane B(2)(TXB(2)), and prostacyclin (measured as 6-keto-PGF(1alpha)) by human term trophoblast cells and villous placental explants. Explants (n=8) and purified trophoblast cells (n=5) were incubated for 24-72 h under either normoxic (21 per cent O(2)) or hypoxic (2 per cent O(2)) conditions. In trophoblast monolayer cultures, hypoxia attentuated PGE(2)production rates to 52+/-9.4 per cent (mean+/-sem, P< 0.05) but recovered to control rates within 48 h. In villous explants, PGE(2)production was significantly decreased after 48 and 72 h of hypoxia versus the normoxic control, accompanied by increased production of 6-keto-PGF(1alpha)to 173.9+/-26.7 per cent after 48 h. TXB(2)production was increased to 172.3+/-25.9 per cent and 653.2+/ 135.7 per cent (P< 0.05) control after 48 and 72 h of hypoxia, respectively. These results were confirmed in villous explants (n=3) cultured in the presence of exogenous 10 microm arachidonic acid. Hypoxia had no significant effect on TXB(2)and 6-keto-PGF(1alpha)in trophoblast cells. In summary, our findings suggest that hypoxia could be responsible for abnormal profiles of prostanoid production commonly observed in women with pre-eclampsia. These results indicate a putative link between hypoxia and compromised placental perfusion. PMID- 11440540 TI - Vasoactive effects of 8-epi-prostaglandin F(2alpha)in isolated human placental conduit and resistance blood vessels in vitro. AB - The effects of 8-epi-prostaglandin F(2alpha)(8-epi-PGF(2alpha)) and the thromboxane A(2)-mimetic U46619 were examined on isolated human fetal placental arteries obtained from normal pregnancies and from those complicated by pre eclampsia. The effects of these agents were examined on both conduit and resistance arteries. 8-epi-PGF(2alpha)was found to be markedly less potent than U46619 in constricting both size vessels. Vasoconstrictor EC(50)s for 8-epi PGF(2alpha)were 4.10x10(-7) m (2.02-8.35x10(-7) m) (mean, 95 per cent CI and 2.05x10(-6) m (0.43-9.89 x10(-6) m) in conduit and resistance arteries, respectively. The maximum vasoconstriction produced by 8-epi-PGF(2alpha)(112+/-17 per cent), (relative to maximum KCl induced vasoconstriction) in conduit vessels was significantly less than that caused by U46619 (152+/-20 per cent). In resistance vessels the maximum vasoconstrictor effects to 8-epi-PGF(2alpha)(208+/ 10 per cent) and U46619 (201+/-19 per cent) were similar, and in both cases significantly greater than the maximal effects seen in conduit vessels. U46619 displayed a similar vasoconstrictor potency in both conduit (EC(50)=1.21x10(-9) m, 0.58-2.51x10(-9) m) and resistance arteries [EC(50)=5.95x10(-9) m, (0.81 43.60x10(-9) m] as was found for 8-epi PGF(2alpha). 8-epi-PGF(2alpha)was equipotent in resistance arteries obtained from women with severely pre-eclamptic pregnancies (EC(50)=1.25x10(-6) m, 0.25-6.17x10(-6) m) compared with normotensive controls. However, the maximum vasoconstrictor effect induced by 8-epi PGF(2alpha)in placental resistance arteries was significantly reduced (99+/-20 per cent) in vessels obtained from severely pre-eclamptic compared with normal pregnancies. These results indicate that 8-epi-PGF(2alpha)displays differential vasoconstrictor activity in the fetal-placental vasculature. Furthermore the vasoconstrictor effects of 8-epi-PGF(2alpha)are reduced in pre-eclampsia, the effect being selective to placental resistance vessels. This reduction may occur as a result of more serious disturbances in the placental microcirculation with the disease process in pre-eclampsia. PMID- 11440541 TI - Effects of maternal iron restriction on placental vascularization in the rat. AB - To investigate the effects of maternal iron deficiency and anaemia on the placenta the composition and vascularization of the placental labyrinth was investigated in iron-restricted rats. Rats in the experimental groups were placed on iron-restricted diets either 1 or 2 weeks before mating and continued on these diets throughout gestation. Placentae were studied at day 21 of gestation. Tissue sections were stained with lectin to allow identification of fetal capillaries and analyzed using stereological techniques. Capillary surface area density and total capillary surface area were decreased in both iron-restricted groups compared with the control group. Capillary length density was decreased in both iron-restricted groups compared with the control group. Total capillary length was significantly reduced in the 1-week, but not in the 2-week, iron-restricted group compared with the control group. Endothelial cell volume was increased in both iron-restricted groups compared to the controls. There were no significant differences in the volume of fetal capillaries, the volume of the maternal blood spaces or the surface area of the maternal-fetal interface between the control and iron-restricted groups. Labyrinthine volume, labyrinthine tissue volume and the surface area of the maternal fetal interface were increased in the 2-week group when compared with the 1-week group. These changes in placental vascularization may contribute to the fetal growth retardation observed in iron restricted litters. PMID- 11440542 TI - Upregulation of neutral endopeptidase expression and enzymatic activity during the differentiation of human choriocarcinoma cells. AB - Neutral endopeptidase (NEP)/CD10, a cell-surface peptidase degrading various bioactive peptides, is mainly present in syncytiotrophoblasts in the human placenta. However, the change in NEP expression upon trophoblast differentiation remains to be clarified. In the present study, we examined the expression of NEP in the differentiating trophoblast using the BeWo choriocarcinoma cell line as a model system. Under the normal culture conditions, NEP was very weakly expressed on most proliferating cytotrophoblastic BeWo cells, while a minority of the cell population (less than 5 per cent ), consisting of giant, multinucleated cells, clearly expressed NEP at the cell membrane. Treatment of BeWo cells with forskolin (FSK) for 48-72 h resulted in an 11- to 44-fold increase in the level of hCG secretion and induced cell fusion leading to the formation of multinucleated syncytiotrophoblasts, indicating functional and morphological differentiation. Fluorescence-activated cell sorting (FACS) analysis revealed that treatment with FSK significantly increased the cell-surface protein expression of NEP on differentiating BeWo cells. Consistently, there was a significant increase in the NEP enzymatic activity after FSK treatment. The level of hCG secretion from the FSK-treated cells was further enhanced when the cells were treated in the presence of the NEP inhibitor phosphoramidon. Immunohistochemical analysis of normal chorionic villi and choriocarcinoma tissues revealed the localization of NEP in syncytiotrophoblastic cells, as opposed to weak or negative staining in cytotrophoblastic cells. These data demonstrate that induction of choriocarcinoma cell differentiation is associated with an increase of NEP/CD10 expression at the cell surface, suggesting a role of this enzyme in regulating differentiated trophoblast functions such as hCG secretion. NEP/CD10 may also be a new cellular differentiation marker of both the normal and neoplastic trophoblast. PMID- 11440543 TI - Flow cytometric measurement of intracellular Th1 and Th2 cytokine production by human villous and extravillous cytotrophoblast. AB - A wide variety of cytokines are present at the maternal-fetal interface, but the extreme cellular complexity of the placenta has made it difficult to determine which cytokines are produced by which cells. Hence novel flow cytometric methods have been applied to determine intracellular cytokine production by specific cell types in placental cell suspensions. Cell suspensions were prepared from first and third trimester chorionic villi and third trimester amniochorion by enzymatic digestion and Percoll density gradient centrifugation. After overnight incubation in the presence of monensin, cells were fixed, permeabilized and labelled with antibodies for villous cytotrophoblast (cytokeratin+, MHC class I-), extravillous cytotrophoblast (cytokeratin+, MHC class 1+) and leucocytes (CD45+). These cell types were further characterized by their expression of EGFR (proliferative cytotrophoblast) and c-erbB2 (invasive cytotrophoblast). Production of IL-4, IL 10, TNF-alpha, IFN-gamma and IL-12 was determined by simultaneous labelling with the appropriate monoclonal antibodies. Only IL-4 was detected consistently in all samples of cytotrophoblast. IL-10 was not detected but IL-10 mRNA was demonstrated in third trimester chorionic villus digests by RT-PCR. Although IL-4 secretion has not been demonstrated, these data suggest that, in vivo there may be a "Th2 type cytokine bias" orchestrated by the trophoblast. It is proposed that other cytokines (including IL-10 and TNF-alpha) are produced by decidual leukocytes, and not cytotrophoblast, at the maternal-fetal interface. PMID- 11440544 TI - Neutrophil activation induced by placental factors in normal and pre-eclamptic pregnancies in vitro. AB - Increased neutrophil activation has been demonstrated in women with pre eclampsia. Activated neutrophils may play a significant role in the vascular endothelial pathophysiology in this disorder of pregnancy. How neutrophils become activated in pre-eclampsia is unknown. It has been proposed that activating factors could be produced and released by the placenta. To test if placental factors could stimulate neutrophil activation and what mechanism might be involved, neutrophils isolated from healthy female volunteers were exposed to the conditioned medium (CM) derived from either normal (Nor) or pre-eclamptic (PE) placental villous culture. Neutrophil-endothelial adhesion, neutrophil superoxide generation, elastase activity and integrin expression were measured. The data were analysed by ANOVA. A P value less than 0.05 was considered statistically significant. All values are expressed as a mean+/-s.e. We found: (1) neutrophil endothelial adhesion was significantly increased in neutrophils exposed PE-CM than those exposed to Nor-CM and non-CM, P< 0.01; (2) both Nor-CM and PE-CM could stimulate neutrophils to generate more superoxide radicals; (3) there was no difference in elastase activity after neutrophil exposure to Nor-CM compared to PE-CM, P> 0.1; (4) significant changes in CD62L and CD11b expression were found in neutrophils exposed to PE-CM. We conclude that factors produced by the placenta can activate neutrophils by an increase in superoxide generation and modulation of adhesion molecule expression. Upregulation of surface adhesion molecule CD11 expression may be responsible for the increased neutrophil endothelial adhesion induced by factors derived from pre-eclamptic placentae. PMID- 11440545 TI - Enhanced protein levels of protein thiol/disulphide oxidoreductases in placentae from pre-eclamptic subjects. AB - Recent studies have indicated that pre-eclampsia is closely associated with oxidative stress both in maternal circulation and in the placenta. Protein thiol/disulphide oxidoreductases, such as thioredoxin, glutaredoxin, and protein disulphide isomerase have recently been found to eliminate reactive oxygen species (ROS) and regenerate oxidatively damaged proteins. Protein thiol/disulphide oxidoreductases may also play a role in combating pre-eclampsia. In this study, we examined the accumulation of 4-hydroxy-2-nonenal (HNE)-modified proteins, which are markers of lipid peroxidation, in human placentae of normal and pre-eclamptic subjects. We also examined the protein levels of thioredoxin, glutaredoxin, and protein disulphide isomerase in placentae. Immunoblotting and immunohistochemistry showed that HNE-modified proteins accumulated to a greater extent in pre-eclamptic placentae than in normal placentae. In both normal and pre-eclamptic placentae, thioredoxin, glutaredoxin, and protein disulphide isomerase were detected in the trophoblasts of the floating villi. The levels of these proteins were increased approximately 2- to 3-fold in the pre-eclamptic placentae compared to the normal placentae. These results indicated that the pre eclamptic placentae were exposed to oxidative stress and that the protein thiol/disulphide oxidoreductases were adaptively induced in pre-eclamptic placentae, suggesting possible roles for thioredoxin, glutaredoxin, and protein disulphide isomerase in protecting placental functions against oxidative stress caused by pre-eclampsia. PMID- 11440546 TI - Expression of intercellular adhesion molecule 1 (ICAM-1) in Plasmodium falciparum infected placenta. AB - We investigated the expression of intercellular adhesion molecule 1 (ICAM-1) in malarial placenta and related histological changes. Thirty-two malarial and 40 control term placentae were collected at Tanga, Tanzania and examined histologically and immunohistochemically. Malaria infected placentae were further divided into acute (15) and chronic (17) cases according to the presence of malarial pigment. The expression of ICAM-1 on monocyte, syncytio- and cytotrophoblasts, endothelial and stromal cells was assessed. Birthweight was lower and leukocyte counts higher in placentae with chronic infection. Many monocytes were present within the intervillous spaces, especially in placentae with chronic infection, and aggregated with parasitized erythrocytes. Some monocytes were adhesive to the surface of fibrinoid deposits. ICAM-1 expression on monocytes of malarial placentae was significantly conspicuous and correlated to the degree of intervillous leukocyte infiltration. Syncytiotrophoblasts often did not show ICAM-1, even though ICAM-1 was expressed by endothelium and weakly by cytotrophoblasts and stromal cells in both infected and control placentae. These results suggest that the expression of ICAM-1 on monocytes contributes to sequestration of infected erythrocytes within the intervillous spaces and their adhesion to fibrin masses and that ICAM-1 is unlikely to be associated with the direct adhesion of infected erythrocytes to the syncytiotrophoblasts. PMID- 11440547 TI - The placenta in pseudoxanthoma elasticum: clinical, structural and immunochemical study. AB - Pseudoxanthoma elasticum (PXE) is a rare genetic disorder clinically characterized by skin, cardiovascular and eye manifestations, mainly due to calcification and fragmentation of elastic fibres. Although infrequent, complications during pregnancy in women affected by PXE have been reported. The aim of the present study was to compare structural features of placentae at term from 14 control and 15 PXE-affected women, in order to better understand if and how abnormal mineral and/or matrix accumulation might affect placental function in PXE. In all cases, pregnancy, fetus growth and delivery were normal. Both gross and light microscopy examination did not reveal dramatic differences between placentae of PXE patients and controls, with regard to weight, dimensions, infarcts, thrombi, inflammatory lesions or vessels. However, necrotic changes and mineralization appeared statistically more pronounced in PXE. By electron microscopy the most remarkable differences between PXE and control placentae were observed in the localization and morphology of mineral precipitates; a significant higher deposition of mineral precipitates was observed associated with the "matrix"-type fibrinoid and among collagen fibrils, especially on the maternal side. Immunocytochemistry revealed the presence of vitronectin and fibronectin associated with the PXE-specific mineralizations and the absence of mineralization on the small and scarce elastic fibres in either controls or in PXE. PMID- 11440548 TI - Placental calcification: a metastatic process? AB - Placental calcification commonly increases with gestational age. The mechanism of apatite mineralization probably involves one of three known mechanisms of tissue calcification: physiological (like bone), dystrophic (ischaemia-related) or metastatic (mineralization in a supersaturated environment). This study was designed to determine the mechanism of calcification by examining (1) the mineral content of placental calcifications in comparison to other physiological and pathological apatites, and (2) the expression of bone morphogenetic proteins (BMPs), which are important in physiological calcification, across gestational age. By energy-dispersive x-ray analysis (EDXA), the Ca/P weight ratio for apatitic mineral from mature calcifications was 2.00+/-0.05 (s.e.), which is similar to that for stones formed in a metastatic, supersaturated environment and lower than that observed in physiological calcification. Biologically active BMP, which was determined by bioassay, was demonstrated in mature and postmature placentae. The BMPs PLAB, PDF and related protein INSL-4 were identified by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), but their mRNA expression was independent of gestational age (7-41 weeks of gestation). We conclude that (1) the identified BMPs were not related directly to placental calcification, which argues against physiological calcification, and (2) the chemical composition of the apatitic mineral was suggestive of rapid formation in a supersaturated environment, which is consistent with a metastatic mechanism of calcification. PMID- 11440549 TI - In-utero characterization of the blood flow in the Hyrtl anastomosis. AB - An anastomosis between the two umbilical arteries, located approximately within 3 cm from the placental insertion, has been previously described at delivery and in utero. However, the prenatal investigation of the Hyrtl anastomosis is limited to case reports. We report a series of antenatal functional evaluations of this vessel. Forty-one women underwent a target ultrasonography to evaluate the blood flow characteristics of the Hyrtl anastomosis. The resistance index of the anastomosis and the umbilical arteries resistance indices before and after the anastomosis were obtained. The direction of the blood flow in the anastomosis was determined by color Doppler evaluation. An anastomosis between the two stems of the umbilical arteries was present in 36 cases while a fusion of the two umbilical arteries was found in the remaining five cases. The median (range) gestational age at diagnosis was 33.1 weeks (25.5-40.1). The median diameter of the anastomosis was 2.3 mm (1.3-7.1). The blood flow in the anastomosis was pulsatile with a median resistance index of 0.62 (0.45-0.85) and unidirectional toward the umbilical artery with lower resistance index. The difference between the resistance indices of the two umbilical arteries was higher after than before the anastomosis [0.07 (0-0.3) versus 0.04 (0-0.17), P=0.05]. The median diameter of the Hyrtl anastomosis was significantly higher when the anastomosis was oblique (n=8) than when it was transverse (n=28) (4.8 mm [2-7.1] versus 2.3 mm [1.3-5.3], P< 0.05). In three out of the five cases with fusion of the two umbilical arteries the placental insertion was marginal or velamentous. We conclude that the Hyrtl anastomosis may act as a pressure-equalizing system between umbilical arteries. This supports the hypothesis that the Hyrtl anastomosis plays an important role when the placental territories supplied by the umbilical arteries are different in size. PMID- 11440550 TI - Three-dimensional ultrasound measurement of the placental volume in early pregnancy: method and correlation with biochemical placenta parameters. AB - Placental size has been an interesting topic of research for many years. The main aim of this study was to investigate the feasibility of measuring the placental volume at the end of the first trimester using three-dimensional (3D) ultrasound and to correlate these volumes to known placental functional indices and to factors affecting the placenta. Women with singleton pregnancies at the end of the first trimester were included into this study. The volume data of the placentae were correlated to the crown-rump length (CRL), placenta-associated plasma protein A (PAPP-A), free beta-human chroangiogonadotropin (f-beta-hCG) and other factors that may affect the placental size or function. A total of 1462 pregnancies could be evaluated. Comparison between CRL and placental volume proved a significant correlation (r=0.43, P< 0.001). Due to the observed proportional growth of CRL and placental volume, a quotient (placental volume/CRL) was calculated for each case. There were no differences between placenta/CRL-quotients in relation to gravidity, parity or smoking. Correlations could be established between the placental volume and PAPP-A and f-beta-hCG (PAPP A: r=0.28, P< 0.001, f-beta-hCG: r=0.10, P< 0.001). The measurement of the placenta in the first trimester can be performed in a high percentage of cases. The placenta/CRL quotient represents a simple method to compare placentae from different gestational days. The correlation between placental volume and maternal serum screening parameters might provide a chance to refine first trimester Down's syndrome serum screening. Future studies will be needed to evaluate the possible clinical use of first trimester placental volume measurements. PMID- 11440551 TI - Placental villus morphology in relation to maternal hypoxia at high altitude. AB - Pregnancy at high altitude is associated with maternal hypoxaemic hypoxia with resultant intervillus blood hypoxia. Maternal haemoglobin concentration and blood gases were measured in pregnant women in two cities in Peru; Lima at sea level (n=18) and Cerro de Pasco at 4300 metres above sea level (n=12). Following delivery, placental sections from both groups were examined histomorphometrically using an image analysis system. Villus diameter, villus cross-sectional area, capillary diameter, capillary cross-sectional area and the percentage of villus cross-sectional area occupied by villus capillaries were calculated and parameters were compared between the two altitude groups. Maternal haemoglobin concentration and maternal blood pH were significantly higher, and maternal pO(2), pCO(2)and O(2)saturation were significantly lower in the high altitude group compared to those at sea level. The villus vessel area as a percentage of villus cross-sectional area and capillary diameter were significantly greater in the cases from the high altitude group and villus vessel area as a percentage of the villus cross-sectional area was significantly related to maternal pO(2)(r= 0.7, P=0.01), and maternal pCO(2)(r=0.7, P=0.02), but multiple regression analysis demonstrated that only pO(2)remained significantly independently associated with these villus histological findings (P=0.03). Placental terminal villi from term pregnancies at high altitude show different morphological features from pregnancies at sea level, and these changes are primarily related to maternal pO(2). The predominant morphological alteration is an increase in villus capillary diameter and therefore of the proportion of villus cross sectional area occupied by capillary lumens. PMID- 11440552 TI - Transfer of recombinant human granulocyte colony stimulating factor (rhG-CSF) from the maternal to the fetal circulation is not dependent upon a functional G CSF-receptor. AB - Administration of granulocyte colony stimulating factor (G-CSF), a haematopoietic growth factor, to pregnant rats increases neutrophil production in the pups. The mechanism for the placental transfer is unknown, but it has been speculated to involve the placental G-CSF receptor (G-CSFR). The purpose of this study was to test that hypothesis. Pregnant mice were treated with a single subcutaneous dose of 50 microg/kg recombinant human G-CSF (rhG-CSF). Mice with an intact G-CSFR ("wild type", WT) and those with a homozygous deletion in the G-CSFR gene (G-CSFR deficient, "knock-out", KO) were studied. At intervals after injection, fetuses were delivered and maternal blood, amniotic fluid (AF) and fetal blood collected. G-CSF concentrations were measured using an enzyme linked immunosorbent assay specific for human G-CSF. Thirty minutes after injection, G-CSF was measurable in the AF (167+/-50 versus 445+/-217 pg/ml, mean+/-sem, WT versus KO) and fetal plasma (774+/-673 versus 427+/-121 pg/ml, WT versus KO). Peak concentrations occurred 2 h after injection in WT dams (572 542+/-41 262 pg/ml) and 4 h in KO dams (616 100+/-96 300 pg/ml). Therefore, in mice, a functional G-CSFR is not essential for the transfer of rhG-CSF from pregnant dams to their fetuses. PMID- 11440553 TI - Cytochemically detectable glucose-6-phosphate dehydrogenase is present in human fetal membrane chorion laeve trophoblasts. AB - We examined the ultrastructural localization of glucose-6-phosphate dehydrogenase (G6PD), a NADPH-generating enzyme, in human fetal membranes at various gestational ages, using newly developed enzyme histochemistry (copper ferrocyanide method). Electron-dense deposits indicative of G6PD activity were clearly visible in the cytoplasm and on the cytosolic side of the endoplasmic reticulum of chorion laeve cytotrophoblasts at various gestational ages. Positive and negative cytochemical controls ensured specific detection of enzyme activity. These observations indicated that chorion laeve trophoblasts were the site of NADPH production. Chorion laeve trophoblast G6PD may play a significant role in fetal membrane physiology, by delivering NADPH to NADPH-dependent oxidoreductases which these cells possess. PMID- 11440554 TI - Intrauterine growth restriction without pre-eclampsia and associated changes in serum inhibin A and activin A levels: a case report. PMID- 11440555 TI - Antihistamines in the treatment of bronchial asthma. Present knowledge and future perspectives. PMID- 11440556 TI - The stem cell factor, its properties and potential role in the airways. PMID- 11440557 TI - Comparison of the anti-bronchoconstrictor activities of inhaled formoterol, its (R,R)- and (S,S)-enantiomers and salmeterol in the rhesus monkey. AB - The principle objective of this study was to define the anti-bronchoconstrictor effects of inhaled racemic formoterol and its (R,R)- and (S,S)-enantiomers in a new model of methacholine-induced bronchoconstriction in the rhesus monkey. A second long-acting beta(2)agonist, salmeterol, was included for comparison. Anaesthetized, spontaneously breathing rhesus monkeys were set up for measuring airway resistance. Blood pressure, heart rate and serum potassium concentrations were measured concomitantly to gauge systemic exposure and the potential for side effects. Formoterol, 0.14, 0.34 and 1.15 microg/kg, administered by aerosol, induced rapidly developing, sustained, dose-related inhibition of the bronchoconstrictor responses to aerosolised methacholine (maximum 76%) accompanied by sustained, dose-related tachycardia. (R,R)-formoterol, 0.56 microg/kg, induced anti-bronconstrictor effects and an associated tachycardia which corresponded closely to the effects seen following twice the dose of the racemate. (S,S)-formoterol, 0.54 microg/kg, was inactive. Salmeterol, 1.4 microg/kg, had no significant anti-bronchoconstrictor effect whereas doses of 5.5 and 30 microg/kg produced quantitatively similar but submaximal anti bronchoconstrictor effects (maximum 47%). Sustained dose-dependent tachycardia was seen with salmeterol over the full dose range. Thus, the anti bronchoconstrictor activity of formoterol resides in the (R,R) enantiomer and the (S,S) enantiomer does not interfere with the activity when present in the racemic form. Furthermore, the data indicate that the present model of methacholine induced bronchospasm in the rhesus monkey could be useful in defining the key properties of beta(2)agonist bronchodilators such as relative potency, efficacy, duration of action and potential for systemic side effects. PMID- 11440558 TI - Concomitant down-regulation of L-arginine transport and nitric oxide (NO) synthesis in rat alveolar macrophages by the polyamine spermine. AB - Polyamines can inhibit NO synthesis in activated macrophages (Mphi). Since NO synthesis in Mphi depends on cellular uptake of L-arginine, effects of polyamines on L-arginine uptake were studied. Rat alveolar Mphi (AMphi) were cultured in absence or presence of lipopolysaccharides (LPS) and/or different polyamines for up to 20 h. LPS increased nitrite accumulation about 10-fold and [(3)H]-L arginine uptake about 2.5-fold, effects almost abolished by spermidine. Spermine had much weaker and putrescine no effects. The effects of spermine depended largely on the presence of serum in the culture medium, suggesting that spermine aldehyde might be involved. Spermine suppressed the mRNA for inducible nitric oxide synthase (iNOS) and that for a specific cationic amino acid transporter (CAT), CAT-2B. In conclusion, in Mphi spermine concomitantly down-regulates NO synthesis and cellular L-arginine uptake by suppressing the expression of iNOS and CAT-2B. By inhibiting specific functions of activated Mphi the polyamine oxidase-polyamine system may play a role as immuno-suppressive modulator. PMID- 11440559 TI - Effect of ET(A) receptor antagonist on pulmonary hypertension and vascular reactivity in rats with congestive heart failure. AB - BACKGROUND: We evaluated the effects of chronic therapy with the selective ET(A)receptor antagonist LU 135252 (LU) on pulmonary vascular reactivity in congestive heart failure. METHODS AND RESULTS: After myocardial infarction (MI) or sham operation, rats were gavaged with LU or saline for 4 weeks. Studies were performed in isolated lungs and to differentiate acute from chronic effects, some experiments were performed with LU in the perfusate. The MI+saline group developed moderate pulmonary hypertension (PH) and right ventricular hypertrophy (RVH). This was improved by LU therapy despite no change in left ventricular function and a larger scar area. Vasodilation to sodium nitroprusside (SNP) was reduced after MI and modestly improved by LU therapy. Vasodilation to acetylcholine (Ach) was similar among the four groups, but accentuated after acute LU administration in the sham+saline and MI+saline groups. A23187 produced higher vasoconstriction (18+/-4%) in the MI+LU compared to the MI+saline (10+/ 3%, P<0.05) and the two control groups (3+/-1% and 4+/-3%, with and without LU, P<0.01): this was reversed to vasodilation following the acute addition of LU. CONCLUSION: ET(A)receptor blockade after MI reduces PH and RVH. LU therapy mildly improves dilation to SNP and favorably modulates pulmonary endothelium-dependent responses. These results support future studies to better define the mechanisms of improvement in pulmonary vascular reactivity after ET receptor antagonist therapy. PMID- 11440560 TI - The placebo response to citric acid-induced cough: pharmacodynamics and gender differences. AB - Characteristics of the response to placebo in a citric acid-induced cough challenge were investigated as part of a randomized, double-blind crossover trial to assess the antitussive effect of dextromethorphan. Baseline cough responses were established on two occasions in 22 healthy subjects. They received 60 ml placebo antitussive syrup and cough frequency following five inhalations of 10% citric acid over 5 min was measured at regular intervals up to 12 h. Response time models of varying complexity were used to describe the placebo cough suppression data. The cough response to placebo was also compared to that of the untreated state. The placebo cough response was best characterized by a non linear increase in cough suppression up to a maximum reduction of 1.6 coughs from baseline at 4-4.5 h, followed by a non-linear return to baseline. The cough response in the untreated state was not different from that of placebo (P=0.99). Females coughed more frequently than males (median number of coughs=10.5 vs. 9.0, respectively P<0.001; Mann-Whitney U test), and adaptation to the cough stimulus was significantly more rapid in females (P<0.025). Accordingly, in trials that use citric acid-induced cough, gender should be considered in study design, particularly in relation to the timing of measurements. PMID- 11440561 TI - Abnormalities of renal endothelin during acute exacerbation in chronic obstructive pulmonary disease. AB - Circulating and urinary levels of endothelin (ET), an endothelium-derived vasoconstrictive and mitogenic peptide have been reported to increase in patients with chronic obstructive pulmonary disease (COPD), but the mechanisms of these abnormalities are not fully understood. Our study objectives were to evaluate pulmonary and renal ET clearance in COPD patients during an acute exacerbation. Our participants included nine consecutive patients with moderate to severe COPD without signs of right heart failure admitted for acute exacerbation and ten healthy volunteers (HV) as controls. ET was detected by radioimmunoassay in venous and arterial blood as well as in a timed urine specimen. For each subject, arterial/venous immunoreactive ET ratio (ir-ETart/ir-ETven) was evaluated as an index of its pulmonary clearance. Creatinine clearance was employed in each case to obtain a corrected renal ir-ET clearance. Glomerular filtration rate (GFR) was also assessed by dynamic(99m)Tc-diethylenetriamine pentaacetic acid renal scintigraphy in six COPD patients during acute exacerbation and at recovery. The ratio ir-ETart/ir-ETven was comparable in COPD patients (0.75+/-0.12) and in HV (0.82+/-0.09). A significant difference was found with respect to 24 h ir-ET urinary excretion between COPD patients during exacerbation as well as at recovery (respectively 142.1+/-12.8 ng/24 h and 89.0+/-15.1 ng/24 h) and HV (65.1+/-10.1 ng/24 h). ET renal clearance was higher in COPD patients than in HV (29.2+/-5.2 ml min(-1)in COPD during exacerbation; 17.5+/-3.9 ml min(-1)at recovery and 13.6+/-2.4 ml min(-1)in HV, P<0.001). GFR was 69.4+/-10.0 ml min( 1)in COPD patients during exacerbation and it significantly increased at the recovery (95.5+/-20.9 ml min(-1)P<0.001). Corrected renal clearance of the peptide was significantly correlated to GFR values during the exacerbation (r= 0.81, P<0.05). Furthermore change in renal ET production resulted associated with changes in paCO(2)(r=0.83, P<0.001) and in paO(2)(r=-0.73, P<0.05). Acute exacerbation in COPD patients causes an increase in renal ET production which is partially reversible at the recovery, in the absence of significant changes in ET 1 circulating levels. ET might contribute to the renal response to hypoxaemia and hypercapnia in COPD. PMID- 11440562 TI - Facile, regioselective [4 + 2] cycloaddition involving 1-aryl-4-phenyl-1 azadienes and allenic esters: an efficient route to novel substituted 1-aryl-4 phenyl-1,4-dihydropyridines. AB - [reaction: see text]1-Aryl-4-phenyl-1-azadienes undergo facile, regioselective [4 + 2] cycloaddition to the C2,C3 pi-bond of allenic esters in refluxing benzene, and the formed adducts undergo a 1,3-H shift to afford novel 2-alkyl-1-aryl-3 ethoxycarbonyl-4-phenyl-1,4-dihydropyridines (78-97%). However, when the reaction is carried at room temperature, besides the [4 + 2] addition, the [2 + 2] mode of addition involving C=N of azadiene and C3,C4 pi-bond of allenic esters also intervenes. The resulting N-aryl-2-ethoxy-carbonyl-methylidene-4-styrylazetidines (17-28%) undergo reorganization on silica gel to afford 2-cyclohexen-1-ones. PMID- 11440563 TI - Relative reactivity of anti- and syn-oximino carbonates and carbamates of 2 pyridylacetic acid esters. AB - [reaction: see text]anti-Oximes of 2-pyridylacetic acid esters are rapidly transformed to pyridine-2-carbonitrile under a variety of conditions while syn oximes bearing tert-butyl esters can be conveniently deprotected to the corresponding carboxylic acid with subsequent fragmentation to the nitrile. PMID- 11440564 TI - Stereoselective trans- and cis-dihydroxylations of 2H-pyranyl and dihydropyridinyl heterocycles synthesized from formal [3 + 3]-cycloaddition reactions of alpha,beta-unsaturated iminium ions with 1,3-dicarbonyl equivalents. AB - [reaction: see text]We describe here an inherent problem in direct epoxidation of the endocyclic olefin in 2H-pyrans fused to 2-pyrones. Such difficulties led to the development of highly stereoselective trans- and cis-dihydroxylations of these olefinic systems in both 2H-pyrans and dihydropyridines fused to a 2 pyrones or a 2-cyclohexenone. Protocols for the removal of the activated allylic hydroxyl group are also reported. PMID- 11440565 TI - A new multicomponent domino reaction of 1,3-dicarbonyl compounds: one-pot access to polycyclic N/O-, N/S-, and N/N-aminals. AB - [reaction: see text] The first multicomponent domino transformation between 1,3 dicarbonyls 1, alpha,beta-unsaturated compounds 2, and functionalized amines 3 is described, providing a one-pot access to fused polyheterocyclic or spiropolyheterocyclic compounds 5 or 6 bearing an aminal function by formation of up to three new cycles, five novel bonds, and up to six stereogenic centers. PMID- 11440566 TI - Nonbasic, room temperature, palladium-catalyzed coupling of aryl and alkenyl iodides with boronic acids mediated by copper(I) thiophene-2-carboxylate (CuTC). AB - [reaction: see text] A new protocol for the palladium-catalyzed, copper-mediated coupling of aryl and alkenyl iodides with boronic acids is described. As an alternative to the well-known and widely used Suzuki cross-coupling, this reaction occurs in the absence of a base at room temperature and should be particularly useful for the construction of substrates bearing base-sensitive and thermally sensitive moieties. PMID- 11440567 TI - Thiadiazole-derived expanded heteroazaporphyrinoids. AB - [structure: see text] Heteroannulenes 1 and 3 containing three subunits of isoindole or pyrrole, respectively, three 1,3,4-thiadiazole moieties, and six aza bridges have been synthesized by reaction of the corresponding diiminoisoindoline or diiminopyrroline with 2,5-diamino-1,3,4-thiadiazole. These conjugated nonaromatic 30 pi-electron macrocycles coordinate three metal ions within their central cavity and constitute one of the rare examples of expanded heteroazaporphyrinoids. PMID- 11440568 TI - The synthesis of an exhaustively stereodiversified library of cis-1,5 enediols by silyl-tethered ring-closing metathesis. AB - [reaction: see text] This report describes the parallel synthesis of all 16 stereoisomers of the cis-1,5 enediol module 1. Compounds 1 derive from 2 by silicon-tethered ring-closing metathesis. Such libraries of stereodiversified ligands provide a unique approach to ligand discovery that employs exhaustive searching of conformational space. PMID- 11440569 TI - Optimization of asymmetric catalysts using achiral ligands: metal geometry induced ligand asymmetry. AB - [reaction: see text] Traditionally, asymmetric catalysts have been optimized by modification of resolved chiral ligands. In this Letter, we optimize the asymmetric addition of diethylzinc to aldehydes by modification of achiral methylene bis(phenol) ligands. Upon coordination of the substrate, the achiral ligand becomes asymmetric, a concept termed Metal Geometry-Induced Ligand Asymmetry. The enantioselectivity of the catalyst formed from a single resolved ligand and several achiral ligands ranged from 9% (R) to 83% (S). PMID- 11440570 TI - Polyquinanes by [4 + 4] cycloaddition-transannular cyclization. AB - [reaction: see text] Photocycloaddition of 2-pyridones yields a rigid polycyclic product containing a 1,5-cyclooctadiene. The cis isomer, with the alkenes in close proximity, undergoes a transannular reaction when treated with chlorine to give a polyquinane product. The chlorination reaction involves migration of an amide nitrogen and forms a single isomer, generating eight stereogenic centers in two steps. PMID- 11440571 TI - A novel cyclooxygenase-inhibitory stilbenolignan from the seeds of Aiphanes aculeata. AB - [structure: see text] Aiphanol (1), a novel stilbenolignan, along with isorhapontigenin (2), piceatannol (3), and luteolin, were isolated by bioassay guided fractionation from the seeds of Aiphanes aculeata Willd. (Arecaceae). The structure of compound 1 was elucidated by spectroscopic methods. Compound 1 is based on an unprecedented stilbenolignan skeleton in which a stilbene moiety is linked with a phenylpropane unit through a dioxane bridge. Compounds 1 and 2 exhibited significant inhibitory activities against cyclooxygenases-1 and -2. PMID- 11440572 TI - Solid-phase rhodium carbenoid reactions: an N-H insertion route to a diverse series of oxazoles. AB - [reaction: see text] The solid-phase synthesis of a series of oxazoles is described. The key step in the construction of these molecules involves the rhodium-catalyzed decomposition of polymer-bound alpha-diazo-beta-ketoesters. These reactions are performed in the presence of primary amides and yield the corresponding N-H insertion products. Subsequent cyclodehydration of these alpha (acylamino)-beta-ketoesters provides the corresponding resin-bound 2,5 disubstituted oxazoles, which are further elaborated during cleavage from the resin. PMID- 11440573 TI - Synthesis of aza bicyclic enones via anionic cyclization: application to the total synthesis of (-)-brunsvigine. AB - [reaction: see text] A general approach to the synthesis of aza bicyclic enones was developed via a simple two-step annulation involving a Mitsunobu protocol and anionic cyclization. According to this strategy the total synthesis of (-) brunsvigine was accomplished with 12% overall yield. PMID- 11440574 TI - Nickel-catalyzed intramolecular homoallylation of omega-dienyl aldehyde. AB - [reaction: see text] In the presence of a catalytic amount of Ni(acac)(2), Et(3)B and Et(2)Zn nicely promote a reductive homoallylic cyclization of omega-dienyl aldehydes and provide 2-allylcyclopentanols and -cyclohexanols with high stereoselectivity in excellent yield. The reaction requires only two kinds of commercially available, inexpensive reagents, Ni(acac)(2) and Et(2)Zn or Et(3)B, and completes at room temperature within 30 min with Et(2)Zn and 1-2 days with Et(3)B. No ligand additives (e.g., phosphine, nitrogen compounds) are required. PMID- 11440575 TI - Chelation, activation, and proximity effects in the deprotection of dithianes with ZnBr(2): applications in the polyketide field. AB - [reaction: see text] Double deprotection of dithiane aldol equivalents is feasible under mild conditions with ZnBr(2) and suitably placed MEM, BOM, and SEM groups which function as protecting, activating, and regiodirecting groups. The procedure is useful in natural product synthesis. PMID- 11440576 TI - First total synthesis of martinellic acid, a naturally occurring bradykinin receptor antagonist. AB - [reaction: see text] The first total synthesis of martinellic acid, a naturally occurring bradykinin receptor antagonist, via a CuI-catalyzed coupling reaction of beta-amino ester 6 with 1,4-diiodobenzene and a guanylation reaction of secondary amine 3 under mild conditions as key steps, is described. PMID- 11440577 TI - Natural glycosides containing allopyranose from the passion fruit plant and circular dichroism of benzaldehyde cyanohydrin glycosides. AB - [structure: see text] Leaves of the edible passion fruit plant, Passiflora edulis, contain benzylic beta-D-allopyranosides 1 and 2, representatives of a rare class of natural glycosides with D-allose as the only sugar constituent. The glycoside 1 is the first known cyanogenic glycoside containing a sugar different from D-glucose attached directly to the cyanohydrin center. Asymmetric perturbation of the (1)L(b) transition of the benzene chromophore was shown to be useful for determination of absolute configuration of the cyanohydrin center of aromatic cyanogenic glycosides. PMID- 11440578 TI - Neutral peptide biradicals formed by dissociative electron transfer. AB - [reaction: see text] A new method for the generation of transient neutral biradicals in liquid solution is reported. Photoinduced electron transfer in aqueous solution of the structures shown above leads to neutral biradicals with peptide spacers. Exchange interactions were measured using time-resolved electron paramagnetic resonance (TREPR) and found to be large for the biradicals possessing two and three glycine residue spacers. These findings are compared to previous results from alkyl-spaced biradicals of similar chain length. PMID- 11440579 TI - Highly diastereoselective aldol additions of a chiral ethyl ketone enolate under Lewis base catalysis. AB - [reaction: see text] The aldol addition of a chiral ethyl ketone enolate bearing an oxygen substituent (OTBS) at the alpha-position proceeds with high internal and relative diastereoselectivities with various achiral aldehydes in good yields. The profound influence of the resident stereogenic center allows for the use of an achiral catalyst, such as HMPA, with minor attenuation in internal stereoselectivity. PMID- 11440580 TI - Efficient and practical synthesis of the A-ring precursor of 19-nor-1alpha,25 dihydroxyvitamin D(3) and its (13)C- or (2)H-labeled derivative. AB - [reaction: see text] The A-ring precursor of 19-nor-1alpha,25-dihydroxyvitamin D(3) (1) and its (13)C- or (2)H-labeled derivative were efficiently synthesized from readily available, optically active 5-(tert-butyldimethylsilyloxy)-2 cyclohexenone (2) through a five-step reaction in 68% overall yield. PMID- 11440581 TI - Kinetically controlled cross-metathesis reactions with high E-olefin selectivities. AB - [reaction: see text] Homoallylic alcohols with anti-allylic substituents display enhanced E-olefin selectivity in cross-metathesis (CM) reactions with allyltrimethylsilane. The high selectivity can be explained via a five-membered chelate intermediate in which the hydroxyl group of the homoallylic alcohol coordinates to the ruthenium metal center of Grubbs' catalyst. PMID- 11440582 TI - Highly efficient synthesis of 1,1-difluoro-2-substituted-1,3-dienes and dienynes from gem-difluorohomoallenyl bromide via palladium-catalyzed cross-coupling reactions. AB - [reaction: see text] A stable gem-difluoroallenylindium 1 is readily transformed to a potentially bifunctional electrophile 2, which can then undergo a transition metal-promoted isomerization-cross-coupling cascade to yield diene 3 and dienyne 4 in very high yields. PMID- 11440583 TI - Cyclization of methyl-substituted 6-heptenyl radicals. AB - [reaction: see text] The behavior of a series of methyl-substituted 6-heptenyl radicals, generated from the corresponding iodides ((Me(3)Si)(3)SiH, AIBN in benzene at 80 degrees C), has been investigated. The stereoselectivity of the 6 exo cyclizations, affording dimethylcyclohexanes, is low, and sizable quantities of methylcycloheptane, generated via 7-endo cyclization, are also produced. PMID- 11440584 TI - Total synthesis of epothilones B and D. AB - [reaction: see text] A highly convergent total synthesis of the natural products epothilone B and D is described. The route is highlighted by efficient generation of a C12-C13 trisubstituted olefin which exploits a sequential Nozaki-Hiyama Kishi coupling and a stereoselective thionyl chloride rearrangement. PMID- 11440585 TI - What is the stabilizing interaction with nucleophilic solvents in the transition state for solvolysis of tertiary derivatives: nucleophilic solvent participation or nucleophilic solvation? AB - [structure: see text] We propose that controversy concerning the mechanism for solvolysis at tertiary carbon is semantic and can be avoided by making a clear distinction between (1) nucleophilic solvent participation, which is stabilization of the transition state for concerted solvolysis by formation of a partial covalent bond to the solvent nucleophile, and (2) nucleophilic solvation, which is stabilization of the transition state for stepwise solvolysis through carbocation or ion pair intermediates by charge-dipole interactions with nucleophilic solvents. PMID- 11440586 TI - Imidazolinium salts as catalysts for the ring-opening alkylation of meso epoxides by alkylaluminum complexes. AB - [reaction: see text] Imidazolinium salts and their N-heterocyclic carbene (NHC) derivatives catalyze the alkylation of a variety of meso epoxides in the presence of triethylaluminum (yield = 70-90%), under mild conditions. Imidazolinium salts are better catalysts than their NHC derivatives but can lead to dimerization side reactions under extended reaction time. Preformed NHC.AlEt(3) complexes and Wanzlick-type olefins, which are dimers of free NHCs, are also catalysts for this reaction. PMID- 11440587 TI - Synthesis of the C1-C12 fragment of fostriecin. AB - [reaction: see text] The synthesis of the C(1)-C(12) fragment of fostriecin was achieved from (S)-glycidol in 15 steps by using an enantioselective allytitanation reaction and a ring-closure metathesis as the key steps. PMID- 11440588 TI - Kinetic resolution in palladium-catalyzed asymmetric allylic alkylations by a P,O ligand system. AB - [reaction: see text] Through the investigation of peptide-based phosphine oxazoline ligands, a simple P,O ligand system was developed. This system provides palladium complexes that are capable of a very high degree of kinetic resolution of 1,3-diphenylprop-2-enyl acetate. The isolated palladium complex was synthesized, characterized, and determined to be an effective procatalyst. PMID- 11440589 TI - N-(tert-butoxycarbonyl)-N-[4-(dimethylazaniumylidene)-1,4-dihydropyridin-1 ylsulfonyl]azanide: a new sulfamyolating agent. Structure and reactivity toward amines. AB - [structure: see text] Synthesis, structure, and reactivity toward amines of the new sulfamoylating reagent 2 are described. Compound 2 allowed sulfamoylation of amines under very mild conditions to give sulfamide derivatives in good yields. PMID- 11440590 TI - Toward the creation of NMR databases in chiral solvents for assignments of relative and absolute stereochemistry: proof of concept. AB - [structure: see text] An NMR database approach in a chiral solvent allows us to predict both relative and absolute stereochemistry of an unknown compound without degradation and/or derivatization. N,alpha-Dimethylbenzylamine (DMBA) is a suitable solvent for this purpose. Using the C.5-C.10 portion of oasomycin A, the feasibility and reliability of this approach is demonstrated. PMID- 11440591 TI - Toward the creation of NMR databases in chiral solvents for assignments of relative and absolute stereochemistry: scope and limitation. AB - [structure: see text] Three additional NMR databases, 1-3, in a chiral solvent are presented. The C.21-C.38 portion of oasomycin A is used to demonstrate the scope and limitation of the universal NMR database approach in a chiral solvent for assignment of relative and absolute stereochemistry without degradation and/or derivatization. PMID- 11440592 TI - Toward the creation of NMR databases in chiral solvents for assignments of relative and absolute stereochemistry: NMR desymmetrization of meso compounds. AB - [structure: see text] Examples of (13)C NMR desymmetrization of meso compounds are presented. On analysis of the NMR profiles of 1,3-diols, the additivity is recognized to predict the NMR profiles of 1,3,5-tirols. PMID- 11440593 TI - Estimating the support of a high-dimensional distribution. AB - Suppose you are given some data set drawn from an underlying probability distribution P and you want to estimate a "simple" subset S of input space such that the probability that a test point drawn from P lies outside of S equals some a priori specified value between 0 and 1. We propose a method to approach this problem by trying to estimate a function f that is positive on S and negative on the complement. The functional form of f is given by a kernel expansion in terms of a potentially small subset of the training data; it is regularized by controlling the length of the weight vector in an associated feature space. The expansion coefficients are found by solving a quadratic programming problem, which we do by carrying out sequential optimization over pairs of input patterns. We also provide a theoretical analysis of the statistical performance of our algorithm. The algorithm is a natural extension of the support vector algorithm to the case of unlabeled data. PMID- 11440594 TI - Stationary bumps in networks of spiking neurons. AB - We examine the existence and stability of spatially localized "bumps" of neuronal activity in a network of spiking neurons. Bumps have been proposed in mechanisms of visual orientation tuning, the rat head direction system, and working memory. We show that a bump solution can exist in a spiking network provided the neurons fire asynchronously within the bump. We consider a parameter regime where the bump solution is bistable with an all-off state and can be initiated with a transient excitatory stimulus. We show that the activity profile matches that of a corresponding population rate model. The bump in a spiking network can lose stability through partial synchronization to either a traveling wave or the all off state. This can occur if the synaptic timescale is too fast through a dynamical effect or if a transient excitatory pulse is applied to the network. A bump can thus be activated and deactivated with excitatory inputs that may have physiological relevance. PMID- 11440595 TI - Vergence dynamics predict fixation disparity. AB - The neural origin of the steady-state vergence eye movement error, called binocular fixation disparity, is not well understood. Further, there has been no study that quantitatively relates the dynamics of the vergence system to its steady-state behavior, a critical test for the understanding of any oculomotor system. We investigate whether fixation disparity can be related to the dynamics of opponent convergence and divergence neural pathways. Using binocular eye movement recordings, we first show that opponent vergence pathways exhibit asymmetric angle-dependent gains. We then present a neural model that combines physiological properties of disparity-tuned cells and vergence premotor cells with the asymmetric gain properties of the opponent pathways. Quantitative comparison of the model predictions with our experimental data suggests that fixation disparity can arise when asymmetric opponent vergence pathways are driven by a distributed disparity code. PMID- 11440596 TI - Topographic independent component analysis. AB - In ordinary independent component analysis, the components are assumed to be completely independent, and they do not necessarily have any meaningful order relationships. In practice, however, the estimated "independent" components are often not at all independent. We propose that this residual dependence structure could be used to define a topographic order for the components. In particular, a distance between two components could be defined using their higher-order correlations, and this distance could be used to create a topographic representation. Thus, we obtain a linear decomposition into approximately independent components, where the dependence of two components is approximated by the proximity of the components in the topographic representation. PMID- 11440597 TI - Blind source separation using temporal predictability. AB - A measure of temporal predictability is defined and used to separate linear mixtures of signals. Given any set of statistically independent source signals, it is conjectured here that a linear mixture of those signals has the following property: the temporal predictability of any signal mixture is less than (or equal to) that of any of its component source signals. It is shown that this property can be used to recover source signals from a set of linear mixtures of those signals by finding an un-mixing matrix that maximizes a measure of temporal predictability for each recovered signal. This matrix is obtained as the solution to a generalized eigenvalue problem; such problems have scaling characteristics of O(N3), where N is the number of signal mixtures. In contrast to independent component analysis, the temporal predictability method requires minimal assumptions regarding the probability density functions of source signals. It is demonstrated that the method can separate signal mixtures in which each mixture is a linear combination of source signals with supergaussian, subgaussian, and gaussian probability density functions and on mixtures of voices and music. PMID- 11440598 TI - A biologically motivated solution to the cocktail party problem. AB - We present a new approach to the cocktail party problem that uses a cortronic artificial neural network architecture (Hecht-Nielsen, 1998) as the front end of a speech processing system. Our approach is novel in three important respects. First, our method assumes and exploits detailed knowledge of the signals we wish to attend to in the cocktail party environment. Second, our goal is to provide preprocessing in advance of a pattern recognition system rather than to separate one or more of the mixed sources explicitly. Third, the neural network model we employ is more biologically feasible than are most other approaches to the cocktail party problem. Although the focus here is on the cocktail party problem, the method presented in this study can be applied to other areas of information processing. PMID- 11440599 TI - A comparative study of feature-salience ranking techniques. AB - We assess the relative merits of a number of techniques designed to determine the relative salience of the elements of a feature set with respect to their ability to predict a category outcome-for example, which features of a character contribute most to accurate character recognition. A number of different neural net-based techniques have been proposed (by us and others) in addition to a standard statistical technique, and we add a technique based on inductively generated decision trees. The salience of the features that compose a proposed set is an important problem to solve efficiently and effectively, not only for neural computing technology but also in order to provide a sound basis for any attempt to design an optimal computational system. The focus of this study is the efficiency and the effectiveness with which high-salience subsets of features can be identified in the context of ill-understood and potentially noisy real-world data. Our two simple approaches, weight clamping using a neural network and feature ranking using a decision tree, generally provide a good, consistent ordering of features. In addition, linear correlation often works well. PMID- 11440602 TI - Time-series analysis as an important contemporary statistical tool. PMID- 11440603 TI - Substance abuse initiatives can help make peace. PMID- 11440604 TI - Aggregate time-series regression in the field of alcohol. AB - Time-series regression of successive aggregate data has been used widely to explore relationships between substance use and harm, or to measure effects of policy interventions. This paper suggests minimal standards to conduct such analyses with respect to validity and reliability of underlying data, statistical techniques and requirements and interpretation of results. PMID- 11440605 TI - Conversation with Charles S. Lieber. PMID- 11440606 TI - Substance abuse policy and peace in the Middle East: a Palestinian and Israeli partnership. AB - This paper describes the working exchange between Israelis, Palestinians and international experts who engaged in a process to promote communications, cooperation and coordination of efforts directed toward peace as well as the prevention and treatment of drug abuse in the region. From 1997 to 1999, a program of training workshops and courses, drug prevention and treatment skills development and collaborative research was conducted on the basis of mutual respect and cooperation among Palestinians and Israelis. By tapping into the issue of substance abuse and by focusing on its professional and academic dimensions, this initiative engaged representative delegations of the police force, academia, treatment centers and various government ministries. While events of this period underscored the dependence of "bottom-up" peace initiatives on the prevailing political situation, the experience revealed the vital role of NGO frameworks in providing a safety net for promoting and sustaining relations as well as addressing an issue of common concern. This case study shows that addiction professionals, both clinicians and researchers, can be instrumental in conflict resolution as well as the prevention and treatment of drug abuse. PMID- 11440607 TI - Meta-analysis without the mantra: a reply to Wilson and "weighted analysis". PMID- 11440608 TI - Alcohol consumption and binge drinking in Novosibirsk, Russia, 1985-95. AB - AIMS: It has been suggested that the effects of alcohol, and binge drinking in particular, contributed to the dramatic fluctuations in Russian mortality rates: rapid decline in 1985-87, slow increase in 1988-91 and sharp increase in 1992-94. To date, there have been no data available to substantiate this claim. We examine for the first time the trends in alcohol intake and binge drinking in a Russian urban population. METHODS: Independent random samples of men and women aged 25-64 in two districts of Novosibirsk city (Western Siberia) have been examined in 1985/86 (1535 men and 1296 women), 1988/89 (1700 men, no women), and 1994/95 (1539 men and 1511 women). Response rates ranged from 71% to 73%. The subjects reported frequency of drinking alcohol, average amount of alcohol consumed at a typical occasion, and their alcohol intake in the week preceding the interview. Two cut-off points to define binge drinking were adopted: > or = 80 g and > or = 120 g at a single occasion. RESULTS: The proportion of men who drank at least once a week increased from 27% in 1985/86 to 38% in 1994/95; corresponding figures among women were 0.6% and 6.5%, respectively. The mean consumption of pure alcohol at a single occasion in men was 90 g in 1985/86, 119 g in 1988/89 and 112 g in 1994/95; in women, it was 33 g in 1985/86 and 32 g in 1994/95. Between the first and the last survey, the mean weekly intake of pure alcohol increased from 120 g to 184 g in men and from 31 g to 41 g in women. Prevalence rates of binge drinking (> or = 80 g at least once a month) in the three surveys were 36%, 52% and 51%, respectively, in men, and 0.4% in the first and 5% in the last survey among women. CONCLUSION: Alcohol consumption and prevalence of binge drinking were high in men and low in women. The frequency of binge drinking among men increased between 1985/86 and 1988/89 and remained stable between 1988/89 and 1994/95. This is not consistent with trends in mortality. Shorter-term fluctuations between surveys, however, cannot be excluded. PMID- 11440609 TI - General Practitioners' views on the English national smoking cessation guidelines. AB - OBJECTIVES: Several countries have followed the US example in publishing national guidelines on smoking cessation interventions. Gauging responses to these guidelines can provide important lessons for future implementation and revision internationally. This study sought to assess General Practitioners' (GPs') views on recently published English smoking cessation guidelines that form the foundation of the UK National Health Service's smoking cessation strategy. DESIGN: Postal survey in which GPs were asked to judge the appropriateness, effectiveness and practicability of key recommendations for primary care in recent national smoking cessation guidelines. SETTING: General practice, England and Wales. SUBJECTS: Random national sample of GPs; 236 GPs completed the questionnaire; effective response rate: 62%. RESULTS: Only 16% of GPs accepted that all the recommendations in the guidelines were appropriate; 43% accepted that it was appropriate to check the smoking status of known smokers when they visit the surgery and only 30% thought it was practicable to advise smokers to stop at every opportunity. However, 77% of GPs thought that they should provide assistance for smokers wanting to stop; 74% believed that they should refer smokers to specialist services if appropriate and a similar proportion (77%) believed that it was appropriate to recommend nicotine replacement therapy. CONCLUSIONS: There was greater acceptance by GPs that they should assist smokers wanting to stop than that they should routinely monitor smoking status and give opportunistic advice to patients to stop. Given that opportunistic advice is a cornerstone of the national guidelines there is a need to find ways of reconciling what is being recommended and what GPs feel is appropriate. PMID- 11440610 TI - Saliva cotinine as indicator of cigarette smoking in pregnant women. AB - OBJECTIVE: To assess use of saliva cotinine as a quantitative objective index of smoking in pregnant women. DESIGN: National survey using quota sampling of pregnant women in England. Face-to-face interviews were conducted in the respondents' own homes. SETTING: Respondents' homes. PARTICIPANTS: N = 1009 pregnant women, mean age = 27.9 years. MAIN OUTCOME MEASURES: Saliva cotinine and current smoking status. RESULTS: Saliva cotinines revealed under-reporting among pregnant women by about 3%. CONCLUSIONS: Pregnant women under-report current smoking status. The optimal cut-off point for validating smoking status among this group requires further research. The results also highlight the need for an improved understanding of the effects of pregnancy on the metabolism of nicotine. PMID- 11440611 TI - Influence of hepatitis C status on alcohol consumption in opiate users in treatment. AB - AIMS: To estimate the prevalence of (a) hazardous and/or harmful drinking, (b) alcohol consumption, (c) perceived hepatitis C status (HCV) in opiate users in treatment and (d) assess the influence of perceived HCV status on consumption and attitudes to risk. DESIGN: Cross-sectional survey. SETTING: A methadone maintenance clinic and a drug treatment centre within a British substance misuse service in London. PARTICIPANTS: A random sample of 93 opiate users in treatment. MEASUREMENTS: Hazardous and/or harmful drinking was assessed using the Alcohol Use Disorders Identification Test (AUDIT). Alcohol consumption was assessed using several indicators. Data on clinical and demographic characteristics, perceived HCV status, change in consumption and attitudes to alcohol consumption were also collected. FINDINGS: A third of the sample were identified as AUDIT cases, 17% drank more than one unit/day and 15% were drinking above the weekly, recommended units for safe drinking (21 for men, 14 for women). Perceived HCV positive status was estimated at 70%. HCV status influenced consumption with fewer HCV positive than HCV negative clients drinking any alcohol in the previous year. Also, more HCV positive clients than HCV negative clients, reduced their consumption after the HCV test result. HCV status had some influence on attitudes to drinking for HCV positive people, although most were aware that abstinence was important for those with HCV positive status. CONCLUSION: Perceived HCV positive status has some influence on alcohol consumption. Despite these findings, training on harm reduction advice on alcohol consumption, particularly in HCV positive clients, should be extended. More intense interventions, within drug treatment services, may be required for those drinkers for whom advice is insufficient. PMID- 11440612 TI - The effect of therapist/patient race- and sex-matching in individual treatment. AB - AIMS: Equivocal findings of the effect that therapist and patient similarity plays in treatment outcome led us to examine the impact of race- and sex-matching on treatment retention and outcome for a sample of people seeking outpatient substance abuse treatment. DESIGN: Patient and therapist characteristics were crossed in a 2 x 2 factorial design. Matching effects were then tested using retrospective data. PARTICIPANTS: Participants were 116 African-American cocaine dependent people. SETTING: The study site was a university-sponsored outpatient treatment facility located in an economically depressed area of a large Northeastern US city. MEASUREMENTS: Follow-up data were drawn from the Addiction Severity Index, the Risk for AIDS Behavior Inventory and a questionnaire measuring post-discharge need for treatment, employment and education. Retention was defined as the number of days inclusive between the last date of service and the date admitted. FINDINGS: No meaningful effects in favor of matching were observed on outcome measures. CONCLUSION: While matching therapists and drug dependent people does not appear to be essential in promoting positive retention and outcome, limitations of this study that include sample size and the homogeneous demographic nature of those seeking therapeutic services at our facility lead us to conclude that replication with a larger number of therapists and patients is needed. In a different therapeutic setting, with a different population presenting for treatment, the mismatching of patients and therapists with respect to factors such as those explored here might well yield differing results. PMID- 11440613 TI - Marijuana Craving Questionnaire: development and initial validation of a self report instrument. AB - AIMS: To develop and validate a multi-dimensional questionnaire on marijuana craving. DESIGN AND MEASUREMENTS: Current marijuana smokers (n = 217) not seeking treatment completed a 47-item Marijuana Craving Questionnaire (MCQ) and forms assessing demographics, drug use history, marijuana quit attempts and current mood. FINDINGS: Exploratory and confirmatory factor analyses indicated that a four-factor solution best described the item structure. Factor subscales derived from the 17 items with significant loadings had respectable internal consistencies and were stable across settings and subgroups. The subscales exhibited low to moderate, positive intercorrelations and were significantly correlated with marijuana use history and a wide range of single-item measures of craving. CONCLUSIONS: Findings suggested that four specific constructs characterize craving for marijuana: (1) compulsivity, an inability to control marijuana use; (2) emotionality, use of marijuana in anticipation of relief from withdrawal or negative mood; (3) expectancy, anticipation of positive outcomes from smoking marijuana; and (4) purposefulness, intention and planning to use marijuana for positive outcomes. These data indicate that the MCQ is a valid and reliable instrument for assessing marijuana craving in individuals not seeking drug abuse treatment and that marijuana craving can be measured in the absence of withdrawal. PMID- 11440614 TI - Patterns of recreational drug use at dance events in Edinburgh, Scotland. AB - AIMS: To describe the patterns of drug use at dance (rave) events in terms of prevalence, frequency, type of drugs used, patterns of use, access and risk associated behaviours. DESIGN: Self-selecting participant-completed survey. SETTING: Three dance events in Edinburgh, Scotland, UK. PARTICIPANTS: One hundred and twenty-two drug users (57% males, 43% females), 90% of whom were in employment or education, with an age range of 16-47, 80% between 18 and 23 years. MEASUREMENTS: Participants who answered 'yes' to the question 'Have you used drugs for dance events in the past year' reported (i) the prevalence, types and frequency of drugs used; (ii) prevalence and contents of mixing drugs; (iii) accessing drugs; and (iv) engagement with drug-associated risk behaviours. FINDINGS: Over 80% of the participants had used ecstasy and amphetamine, over 30% cocaine and LSD; over 10% nitrites, psilocybin and ketamine and less than 5% had used crack or tranquillizers. Participants reported regular consumption of ecstasy and amphetamine (e.g. 35% used ecstasy and 25% amphetamine on a weekly basis) often taken in combination, with the occasional use of cocaine, LSD, ketamine and psilocybin. Poly- and mixing-drug behaviours were significantly more likely than monodrug usage. Drugs were accessed through friends than from any other source. Eighty-five per cent reported mixing drugs and/or alcohol, 35% driving on drugs, 36% having a bad experience on drugs; 30% unprotected sex; and 0.9% injecting drugs. Women in the sample reported higher consumption than men. CONCLUSIONS: Dance-drug use has a characteristic pattern that has implications for health promotion and criminal policy. PMID- 11440615 TI - Needle fixation, the drug user's perspective: a qualitative study. AB - AIMS: Changing drug users' injecting behaviour is central to the project of drug services. Information about the idea of "needle fixation" is fragmentary and of uncertain relevance to contemporary injecting drug users. The aim of the study is to describe injecting drug users' ideas about needle fixation. PARTICIPANTS AND DESIGN: Twenty-four participants, some of whom identified themselves as needle fixated, were recruited from four drugs agencies in south Wales. Participants took part in semi-structured interviews, recorded, transcribed and qualitatively analysed using Atlas/ti software. FINDINGS: Participants describe a range of behaviours and experiences which fit with previous ideas of needle fixation, including ritualization, substitution of other drugs, injection of water and associations with deliberate-self-harm and sex. Participants describe high levels of needle aversion and add detail to previous partial descriptions of needle fixation. CONCLUSIONS: Issues conveniently considered together as needle fixation are current among injecting drug users and may be relevant to the inability of some drug users to change from injecting drug use. PMID- 11440616 TI - Adolescent emergency department presentations with alcohol- or other drug-related problems in Perth, Western Australia. AB - AIMS: To identify the morbidity, type of substance used and the pattern of presentation by adolescents with problems related to alcohol or other drug (AOD) use. DESIGN: A 4-week retrospective review of hospital records. SETTING: Four metropolitan hospitals in Perth, Australia. PARTICIPANTS: There were 1064 presentations by people aged 12-19 years of which 160 (15%) were related to AOD use. The median age of the AOD cases was 17 (interquartile range 16-19) of whom 97 (61%) were male and 19 (12%) were Indigenous Australians. FINDINGS: Alcohol was the most frequent precursor to presentation (66, 41%) followed by heroin (24, 15%) and prescription/over-the-counter drugs (24, 15%). Injury was the most common diagnosis at presentation (50, 31%), followed by overdose/drug use (47, 29%). A diagnosis of injury was significantly more likely following the use of alcohol than other categories of substances (chi(2) = 42.07, df = 3, p < 0.001). Deliberate self-harm (DSH) occurred in more female than male cases (chi2 = 7.4, df = 1, p < 0.01). Presentations were more frequent over the weekend (102, 64%) than on weekdays, and the length of stay was significantly shorter for weekend cases (Mann-Whitney U 2132, p < 0.05). CONCLUSIONS: Given the small window of opportunity to provide AOD treatment to youth following hospital presentation, a number of suggestions are made. From a harm-minimization perspective the focus of interventions should be on alcohol use by male youth and DSH associated with prescription/over-the-counter drug use by female adolescents. In addition, Indigenous youth are over-represented in hospital presentations, but there is currently a lack of evaluated interventions designed for them. PMID- 11440617 TI - Cost-effective treatment in Brazil and elsewhere. PMID- 11440618 TI - An improved vector for high-level, consistent retroviral transgene expression in human thymocytes after competitive reconstitution from transduced peripheral blood stem cells. AB - One problem in hematopoietic stem cell (HSC)-based gene therapy is the low-level, and often transient, transgene expression in progeny cells in vivo. Here we have evaluated retroviral vector designs for improved long-term in vivo transgene expression levels in thymocytes recovered after transplantation of gene-modified HSCs. First, several vector designs were screened in vitro by single-cell analysis of transgene marking and expression to rapidly identify optimal vectors for sensitive tracking of marked cells. Next, using one optimal vector, we show that gene-modified HSCs can competitively reconstitute thymopoiesis in SCID-hu thymus/liver mice, with transgene expression detectable on 0-40% of marked donor thymocytes. Modified vector designs (termed MSCV-SAR and MoMLV-SAR), which enhance transgene expression in primary T cells in vitro, were shown here to improve in vivo transgene expression levels per cell 12- to 14-fold (mean fluorescence intensity was 2175 for MSCV-SAR vs. 174 for LNGFRSN; %NGFR(+) donor(+) cells with high-level expression was 58% for MSCV-SAR vs. 4% for LNGFRSN). Importantly, 61% of grafts had high-level transgene expression on thymocytes with the MSCV-SAR vector versus 0% of grafts for LNGFRSN or MoMLV-SAR. Transgene expression was demonstrated in various stages of thymocyte differentiation and was consistently detected in early thymic progenitors. We suggest that the MSCV-SAR vector described here is particularly advantageous for applications requiring high-level, consistent transgene expression in a diverse repertoire of T cells derived from gene-modified HSC grafts. PMID- 11440619 TI - Cross-strain protection against clinical and laboratory strains of Pseudomonas aeruginosa mediated by dendritic cells genetically modified to express CD40 ligand and pulsed with specific strains of Pseudomonas aeruginosa. AB - We have shown that dendritic cells (DCs) genetically engineered with a recombinant adenovirus vector (Ad) to express CD40 ligand (CD40L) elicit specific humoral immunity against the Pseudomonas aeruginosa laboratory strain PAO1, without CD4(+) T cell help. In the present study, using several different strains of P. aeruginosa, we examine whether this strategy is generally applicable to enhancing clinically relevant pathogen-specific immunity. Mice immunized with DCs modified with CD40L and pulsed with heat-killed P. aeruginosa clinical strain PA514, originally isolated from the sputum of an individual with cystic fibrosis, survived lethal respiratory challenge with PA514-impregnated agar beads. Consistent with this effective in vivo protection, the immunized mice generated high levels of serum isotype-switched antibodies directed against PA514 without concomitant nonspecific elevations of total serum immunoglobulin levels. The CD40L genetically engineered DCs pulsed with seven of eight different strains of P. aeruginosa afforded significant, albeit variable, cross-protection against lethal respiratory challenge with a clinical (PA514) or laboratory (PAO1) strain of P. aeruginosa. CD40L genetically modified DCs pulsed with a clinical (PA514) or laboratory (PAO1) strain of P. aeruginosa initiated cross-reacting antibody responses against each other, but not against Escherichia coli and vice versa. These observations may be useful in developing vaccines for infectious diseases, including P. aeruginosa infection. PMID- 11440620 TI - Intratumoral delivery of interleukin 12 expression plasmids with in vivo electroporation is effective for colon and renal cancer. AB - We report on an antitumor treatment involving electrogene therapy (EGT), a newly developed in vivo gene transfer method using electroporation. We carried out in vivo EGT in a subcutaneous model of CT26 colon carcinoma cells, using plasmid DNAs encoding interleukin 12 (IL-12) subunits. For this purpose, we developed two IL-12 expression systems: a cotransfer system using a plasmid encoding the IL-12 p40 subunit and a plasmid encoding the IL-12 p35 subunit, and a single-vector system using a plasmid expressing a p40-p35 fusion protein. Both transfer systems significantly inhibited the growth of CT26 tumor. Immunohistochemical analysis of IL-12 EGT-treated tumors revealed enhanced infiltration of CD8(+) cells into the tumor tissue, while reverse transcriptase-polymerase chain reaction confirmed the increased expression of interferon gamma within treated tumors. The same IL-12 EGT applied to the nude mouse model was not effective, suggesting the critical role of T cell infiltration in this treatment. The inhibitory effects revealed in experiments in which previously treated mice were rechallenged with a second inoculation of CT26 tumor cells suggested that IL-12 EGT may also establish partial systemic antitumor immunity. The growth of IL-12 EGT-treated Renca tumors, a renal cell carcinoma, was also significantly inhibited. These findings suggest that EGT of the IL-12 gene has the potential to be an effective anticancer gene therapy. PMID- 11440621 TI - Lesion-targeted injectable vectors for vascular restenosis. AB - Pathologic lesions caused by catheter-based revascularization procedures for occlusive artery disease include disruption of the endothelium, exposure of extracellular matrix (ECM) proteins, and proliferation of vascular smooth muscle cells, which lead to neointima formation and restenosis. We have developed matrix collagen-targeted retroviral vectors that are able to accumulate at sites of vascular injury (Hall et al., Hum. Gene Ther. 1997;8:2183-2192; Hall et al., Hum. Gene Ther. 2000;11:983-993). The primary tissue-targeting motif, adapted from the physiological surveillance sequence found in von Willebrand factor, served to localize and concentrate the vector within vascular lesions. In the present study, we evaluated the efficiency of this vector-targeting system in rats with nonligated balloon-injured carotid arteries. Both intraarterial (by retrograde femoral artery catheterization) and intravenous (via femoral vein) injection of a matrix-targeted vector enhanced transduction of neointimal cells ( approximately 20%) at severely denuded areas when compared with the nontargeted vector (<1%). Further, intraarterial instillation of a matrix-targeted, but not a nontargeted, vector bearing an antisense cyclin G1 construct inhibited neointima lesion formation in the injured carotid arteries. Taken together, these data indicate that strategic targeting of retroviral vectors to vascular lesions would have therapeutic potential in the management of vascular restenosis and many other disorders of uncontrolled proliferation where endothelial disruption, ECM remodeling, and collagen deposition form the nexus for preferential vector localization and concentration in vivo. PMID- 11440622 TI - Cytokine gene therapy for myocarditis by in vivo electroporation. AB - Cytokines are important pathophysiologic and pathogenic factors in cardiovascular disorders, including viral myocarditis. We attempted to treat viral myocarditis with cytokine gene therapy by transferring an inhibitory cytokine, IL-1 receptor antagonist (IL-1ra) or viral IL-10 (vIL-10), by in vivo electroporation, a new method for gene transfer into muscle. Four-week-old male DBA/2 mice were inoculated intraperitoneally with 10 PFU of encephalomyocarditis virus. Immediately after virus inoculation, an expression plasmid carrying IL-1ra or vIL 10 was injected into tibialis anterior muscles followed by electroporation. Serum levels of IL1ra and vIL-10 reached 10.5 and 2.3 ng/ml, respectively, on day 5, when gene expression reached its peak. Histopathological examination showed that myocardial cellular infiltration was improved in mice treated with IL-1ra or vIL 10 compared with the control group. On day 14 after the onset of myocarditis, transfer of IL1ra or vIL-10 expression plasmid had significantly improved the survival rates of the animals. The expression of TNF-alpha was decreased to 0.60 fold (p < 0.005) and inducible nitric oxide synthase (iNOS) 0.43-fold (p < 0.005) by IL-1ra treatment, and the expression of IFN-gamma in the heart was decreased to 0.35-fold (p < 0.05), and iNOS 0.21-fold (p < 0.005), by vIL-10 relative to the controls. These results show that gene therapy with IL-1ra or vIL-10 expression plasmid was effective in the treatment of viral myocarditis, and in vivo electroporation may be a useful method by which to deliver cytokine therapy in cardiovascular diseases. PMID- 11440623 TI - Inhibition of angiogenesis by adenovirus-mediated sFlt-1 expression in a rat model of corneal neovascularization. AB - Pathological angiogenesis, or the production of new capillary vessels from preexisting vasculature, within the eye is a serious event that often leads to blindness. Upregulation of vascular endothelial growth factor (VEGF) has been linked to neovascularization in the eye, suggesting that it could be a suitable target to inhibit angiogenic changes. This work investigated whether the presence of a proven antiangiogenic factor, the soluble variant of the VEGF receptor, sFlt 1, in the anterior chamber is sufficient to inhibit new vessel formation in the cornea in an animal model of corneal neovascularization. A recombinant adenovirus vector that can mediate efficient in vivo gene transfer and expression in ocular cells was selected as a delivery agent. We have shown that after the injection of Ad.betagal into the anterior chamber of normal and cauterized rat eyes, corneal endothelial cells and cells of the trabecular meshwork were efficiently transduced and that beta-galactosidase (beta-Gal) expression was maintained up to 10 days postinjection. Cauterization significantly increased the amount of immunoreactive VEGF in vehicle- or Ad.null-injected animals (t test, p < 0.001 and p < 0.001, respectively). However, when cauterization was combined with Ad.sflt injection there was no statistically significant increase in the amount of immunoreactive VEGF (p = 0.12). The injection of Ad.sflt into the anterior chamber slowed or inhibited VEGF-induced angiogenic changes. After cauterization, 100% of uninjected and vehicle-injected and 82% of Ad.null-injected animals developed moderate to severe corneal angiogenesis in contrast to 18% of Ad.sflt injected animals. These in vivo results suggest that the transient presence of antiangiogenic agents in the anterior chamber can be successfully used to inhibit the development of corneal angiogenesis. PMID- 11440624 TI - Systemically expressed soluble Tie2 inhibits intraocular neovascularization. AB - Retinal and choroidal neovascularization are the most frequent causes of severe and progressive vision loss. Studies have demonstrated that Tie2, an endothelial specific receptor tyrosine kinase, plays a key role in angiogenesis. In this study, we determined whether adenovirus-mediated gene delivery of extracellular domain of the Tie2 receptor (ExTek) could inhibit experimental retinal and choroidal neovascularization. Immunofluorescence histochemistry with a monoclonal antibody to human Tie2 showed that Tie2 expression is prominent around and within the base of newly formed blood vessels of retinal and choroidal neovascular lesions. A single intramuscular injection of adenovirus expressing ExTek genes achieved plasma levels of ExTek exceeding 500 microg/ml in mice for 10 days (in neonates) and 7 days (in adults). This treatment inhibited retinal neovascularization by 47% (p < 0.05) in a murine model of ischemia-induced retinopathy. The same treatment reduced the incidence and extent of sodium fluorescein leakage from choroidal neovascular lesions by 52% (p < 0.05) and 36% (p < 0.01), respectively, in a laser-induced murine choroidal neovascularization model. The same mice showed a 45% (p < 0.001) reduction of integrated area of the choroidal neovascularization. These findings indicate that Tie2 signaling is a common component of the angiogenic pathway in both retinal and choroidal neovascularization, providing a potentially useful target in the treatment of intraocular neovascular diseases. PMID- 11440625 TI - Wild-type adenovirus decreases tumor xenograft growth, but despite viral persistence complete tumor responses are rarely achieved--deletion of the viral E1b-19-kD gene increases the viral oncolytic effect. AB - Strategies to target viral replication to tumor cells hold great promise for the treatment of cancer, but even with replicating adenoviruses complete tumor responses are rarely achieved. To evaluate replicating adenoviral vectors, we have used A549 human lung cancer nude mouse xenografts as a model system. Intratumoral injection of wild-type adenovirus (Ad309) significantly reduced tumor growth from day 14 (p = 0.04) onward; however, tumor volumes reached a plateau at day 50. At 100 days, high levels of titratable virus were present within persistent viable tumors. In contrast to viral injection into established tumors, when tumor cells were infected in vitro with wild-type virus and then mixed with uninfected tumor cells, 1% of infected cells was sufficient to prevent tumor establishment. An E1b-19kD-deleted viral mutant (Ad337) was more efficient than Ad309 in this cell-mixing model. Just 1 cell in 1000 infected with Ad337 prevented tumor growth. However, although better than wild-type virus, Ad337 was unable to eradicate established flank tumors. These data suggest that although replicating adenoviruses exhibit significant oncolytic activity, barriers within the established tumor, such as connective tissue and tumor matrix, may limit the spread of virus. Strategies to enhance viral spread through established tumors are therefore likely to greatly improve the therapeutic efficacy of replicating adenoviruses. PMID- 11440626 TI - Hydroxychloroquine enhances the endocrine secretion of adenovirus-directed growth hormone from rat submandibular glands in vivo. AB - Use of gene transfer technology for treating single protein deficiency disorders requires delivery of therapeutic levels of the transgene product. We have suggested that salivary glands may provide a potentially valuable target site for certain systemic applications of gene therapeutics (He et al., Gene Ther. 1998;5:537-541). However, the ability of salivary glands to deliver therapeutic proteins to either the upper gastrointestinal tract via saliva or to the bloodstream, as required, must be carefully evaluated. In the anterior pituitary gland, human growth hormone (hGH) is secreted into the bloodstream via the regulated secretory pathway. However, when expressed from an adenoviral vector delivered to salivary glands, most hGH follows the regulated, tissue-specific, exocrine secretory pathway into saliva, where it is not therapeutically useful. We tested the hypothesis that the commonly used, FDA-approved drug hydroxychloroquine (HCQ) can divert adenovirus-directed hGH from this regulated secretory pathway in rat submandibular glands and enhance delivery into the bloodstream. In untreated rats, there was approximately 20-fold more vector directed hGH in saliva than in serum. Administration of HCQ led to a shift of hGH secretion into the bloodstream. When delivered at doses of 1 or 10 mg/kg body weight, via intraperitoneal injection plus intraductal infusion, the saliva:serum hGH ratio was approximately 2:1. Such HCQ delivery did not significantly alter the total amount of hGH measured, but increased the serum level of hGH 5- to 6 fold. Also, HCQ had no significant effects on serum chemistries or hematological parameters. We conclude that HCQ is able to significantly enhance hGH secretion from salivary glands into the bloodstream and may be useful to facilitate clinical applications of gene therapeutics via salivary glands. PMID- 11440627 TI - Induced apoptosis supports spread of adenovirus vectors in tumors. AB - Selectively replicating viruses hold promise as anticancer agents. To eliminate the tumor, these viruses must efficiently spread throughout the tumor and induce oncolysis. We hypothesized that viral release and spread could be supported by apoptosis induced after assembly of de novo-produced virions in tumor cells. As a model to test this, we employed an adenovirus vector that replicated in human tumor cell lines. Expression of a dominant-negative I-kappaB from this vector sensitized tumor cells to recombinant human tumor necrosis factor alpha (TNF alpha)-mediated apoptosis. We found that apoptosis induced during viral DNA replication compromised virus production, whereas apoptosis induced after virion assembly enhanced viral release from infected cells and dissemination. Electron microscopy demonstrated that viral particles were associated with or included in apoptotic bodies whose phagocytosis by neighboring cells provides a potential means for viral spread. Apoptosis induced after viral replication also supported spread in vivo, in subcutaneous tumors or liver metastases, resulting in a delay of tumor growth. Our findings could be applicable to other selectively replicating viruses or antitumor strategies that involve application of proapoptotic or cytolytic agents. PMID- 11440628 TI - A genetic immunization adjuvant system based on BVP22-antigen fusion. AB - DNA vaccines must induce a greater immune response to be effective in the biomedical industry. Therefore, we tested the trafficking trait of the bovine herpesvirus 1 (BHV-1) protein VP22 (BVP22) fused to an antigen and applied this unique trait to genetic immunization. DNA immunization with BVP22-antigen stimulates immune responses superior to that of standard DNA immunization. Mice were injected intramuscularly with gene constructs expressing the antigen yellow fluorescent protein (YFP), YFP fused to BVP22, or YFP fused to BHV-1 tegument protein VP16 (BVP16). The results revealed a significantly enhanced YFP antibody response with BVP22-YFP DNA immunization compared with either YFP or BVP16-YFP gene immunization. Notably, the BVP22-YFP DNA construct induced a stronger T helper 1 (Th1) response, based on IFN-gamma and IL-4 cytokine levels, and IgG2a/IgG1 ratios. Furthermore, BVP22-YFP genetic immunization induced a greater cytotoxic T lymphocyte response. The genetic adjuvant properties of BVP22 can make DNA vaccines much more effective clinically. PMID- 11440629 TI - Gene therapy clinical trials in Belgium. PMID- 11440631 TI - Differential properties of two putative nuclear localization sequences found in the carboxyl-terminus of human ifn-gamma. AB - Interferon-gamma (IFN-gamma), a protein that uses the Jak-Stat pathway for signal transduction, translocates rapidly to the nucleus in cells treated extracellularly with the cytokine. A nuclear localization sequence (NLS) has been identified and characterized in the C-terminus of IFN-gamma. Both human and murine IFN-gamma contain this NLS. We show in this report that human IFN-gamma (HuIFN-gamma) contains a second NLS at an upstream site, as determined in standard import assays using digitonin-permeabilized HeLa cells. The primary sequence, analogous with the NLS sequence identified in murine IFN-gamma, representing amino acids 122-132 of HuIFN-gamma was capable of mediating the nuclear import of the autofluorescent protein allophycocyanin (APC) in an energy dependent manner. The second sequence, representing amino acids 78-92 of HuIFN gamma, was also capable of mediating the nuclear import of APC in an energy dependent manner but to a greatly reduced extent. The nuclear import of both sequences conjugated to APC was strongly blocked by competition with unconjugated HuIFN-gamma(122-132). Competition by the sequence HuIFN-gamma(78-92) effectively blocked the import of APC-conjugated HuIFN-gamma(78-92) but, at the same concentration, was not capable of inhibiting the nuclear import of APC-conjugated HuIFN-gamma(122-132), suggesting that HuIFN-gamma(78-92) was a less efficient NLS than HuIFN-gamma(122-132). This is consistent with >90% loss of antiviral activity of HuIFN-gamma lacking the downstream NLS in 122-132. The nuclear import of APC-conjugated HuIFN-gamma(122-132) was inhibited by a peptide containing the prototypical polybasic NLS of the SV40 T NLS, which suggests that the same Ran/importin cellular machinery is used in both cases. PMID- 11440632 TI - Redox imbalance in Crohn's disease intestinal smooth muscle cells causes NF kappaB-mediated spontaneous interleukin-8 secretion. AB - Interleukin-8 (IL-8), a chemokine secreted by cells at injury sites, has recently been recognized as involved in the pathogenesis of Crohn's disease. However, the pathogenesis of enhanced spontaneous transcription of IL-8 by the bowel in patients with Crohn's disease is undefined. Although IL-8 is secreted primarily by neutrophils, macrophages, and endothelial and epithelial cells, we observed the involvement of mesenchymal cells in the inflammatory process. A smooth muscle cell line isolated from the ileum of a patient with Crohn's disease (CDISM) and maintained in culture exhibited spontaneous transcription and secretion of IL-8 when compared with intestinal smooth muscle cells obtained from a normal subject (NHISM). Furthermore, IL-8 transcription from CDISM cells was associated with remarkable spontaneous activation of the oxidant-sensitive transcription factor NF-kappaB, as assessed by transient transfection assays with an IL-8 promoter reporter construct, Western blot analysis, and electrophoretic mobility shift assays (EMSA). Finally, we report here that CDISM cells exhibit significantly altered redox balance. The antioxidant pyrrolidine dithiocarbamate (PDTC) restored the redox equilibrium by mechanisms that inhibit binding of NF-kappaB to its cognate site on the IL-8 promoter. These findings suggest that restoration of the redox balance could hold promise for therapeutic intervention in Crohn's disease. PMID- 11440633 TI - Canine interleukin-5: molecular characterization of the gene and expression of biologically active recombinant protein. AB - Interleukin-5 (IL-5), which is produced primarily by type 2 T helper lymphocytes (Th2), is an eosinophil differentiation and activation factor. Increased numbers of eosinophils in peripherial blood or tissues (eosinophilia) are observed in asthmatic human patients, in animals with helminth infections, and in dogs with allergic diseases. Antagonism of IL-5 activity is being explored as a potential treatment of a number of disease conditions associated with eosinophils in animal models. In order to study the expression and function of this cytokine in the dog, we have isolated and characterized the canine IL-5 gene. The canine IL-5 polypeptide deduced from the cDNA is composed of 134 amino acids that share varying degrees of homology with IL-5 isolated from several mammals. The genomic structure of the canine IL-5 gene consists of four exons and three introns in the coding region, similar to that of the previously characterized human and mouse IL 5 genes. Recombinant canine IL-5 protein, expressed in Pichia pastoris, is biologically active in a cell proliferation assay. Canine IL-5 gene sequences and the biologically active protein described in this study will be useful reagents for future studies of this cytokine in physiologic processes and in pathologic conditions of the dog. PMID- 11440634 TI - Involvement of protein phosphatase 2A in the interleukin-3-stimulated Jak2-Stat5 signaling pathway. AB - In this study, we report that the tyrosine kinase, Janus kinase 2 (Jak2), associates with the serine/threonine protein phosphatase 2A (PP2A) in 32Dcl3 myeloid progenitor cells. The association between Jak2 and PP2A transiently increases following interleukin-3 (IL-3) stimulation and activation of Jak2. The catalytic subunit of PP2A is tyrosine phosphorylated by Jak2 in vitro and in vivo, resulting in inhibition of phosphatase activity. PP2A also associates with Stat5 in 32Dcl3 cells in an IL-3-dependent manner. Pretreatment of 32Dcl3 cells with okadaic acid (OA), an inhibitor of PP2A, resulted in increased tyrosine phosphorylation and nuclear translocation of Stat5. Our results suggest that PP2A plays a negative regulatory role in regulating the IL-3 signaling pathway via formation of complexes with Jak2 and Stat5. PMID- 11440635 TI - The interferon-inducible Stat2:Stat1 heterodimer preferentially binds in vitro to a consensus element found in the promoters of a subset of interferon-stimulated genes. AB - Regulated expression of type I interferon (IFN)-stimulated genes (ISG) requires the binding of the signal transducer and activator of transcription (Stat) complexes to enhancer elements located in the ISG promoters. These enhancer elements include the IFN-stimulated response element (ISRE) and the palindromic IFN-gamma activation site (GAS) element. Regulated expression of ISRE containing ISG depends on IFN-stimulated gene factor 3 (ISGF3), a heterodimer involving Stat1 and Stat2 in association with a DNA-binding adapter protein, p48/IFN regulatory factor-9 (IRF-9). Several GAS binding Stat complexes involving Stat1, Stat3, Stat4, and Stat5 have been described, but their contribution to GAS dependent ISG expression remains to be established. We and others previously identified an IFN-alpha-inducible Stat2:1 heterodimer that exhibits binding to the GAS element of the IRF-1 gene. These previous studies raise the possibility that Stat2:1 may participate in the transcriptional activation of the subset of ISG containing GAS elements. To address this question, we performed a PCR assisted binding site selection procedure to define the Stat2:1 recognition sequence. The data reveal that Stat2:1 preferentially binds to a palindromic sequence similar to the consensus GAS element found in the promoter of several ISG. Our results establish that in addition to the classic complex formation involving Stat2, Stat1, and p48 associations, Stat2:1 heterodimers are formed in response to IFN treatment that may play an important role in the transcriptional regulation of certain ISG. PMID- 11440636 TI - An anti-inflammatory drug, propagermanium, may target GPI-anchored proteins associated with an MCP-1 receptor, CCR2. AB - Monocyte chemoattractant protein-1 (MCP-1) promotes the migration and activation of monocytes and plays a pivotal role in the development of chronic inflammation. Propagermanium (3-oxygermylpropionic acid polymer) has been used as a therapeutic agent against chronic hepatitis B in Japan. We report here that propagermanium specifically inhibits in vitro chemotactic migration of monocytes by MCP-1. Propagermanium did not inhibit binding of MCP-1 to a human monocytic cell line, THP-1 cells, or affect intracellular Ca(2+) mobilization or the cAMP concentration in MCP-1-treated THP-1 cells. The effect of propagermanium seems to require glycosylphosphatidylinositol (GPI)-anchored proteins, as cleavage of GPI anchors by phosphatidylinositol-phospholipase C (PI-PLC) eliminated the inhibitory activity of propagermanium. Anti-GPI-anchored protein antibodies, such as anti-CD55 and anti-CD59, reduced staining of C-C chemokine receptor 2 (CCR2) with an anti-CCR2 antibody against the N-terminus of CCR2 in a flow cytometric analysis, and these antibodies also selectively inhibited MCP-1-induced migration of THP-1 cells. Furthermore, under fluorescence microscopy, GPI-anchored proteins colocalized with CCR2 on THP-1 cells. These results suggest that propagermanium may target GPI-anchored proteins that are closely associated with CCR2 to selectively inhibit the MCP-1-induced chemotaxis, thus providing a mechanistic basis for the anti-inflammatory effects of the drug. PMID- 11440637 TI - Selective expression of nonsecreted interferon by an adenoviral vector confers antiproliferative and antiviral properties and causes reduction of tumor growth in nude mice. AB - Replication-deficient adenoviruses expressing human interferon-alpha2b (HuIFN alpha2b) or the hybrid IFN-alpha2alpha1 or those with the secretory signal deleted, whose express is driven by the alpha-fetoprotein (AFP) promoter, were constructed and characterized. Synthesis of IFN protein and secretion or intracellular retention were tested by Western blotting and immunoassay. Expression of IFN by the recombinant adenoviruses was restricted to cells that constitutively express AFP. In these cells, expression of both secreted and nonsecreted recombinant IFN resulted in inhibition of cell proliferation, resistance to viral infection, induction of major histocompatibility complex (MHC) class I expression, increased apoptosis, and activation of an IFN stimulated response element (ISRE)-containing promoter. Also, the induction of protein kinase R (PKR), increased phosphorylation of Stat1, and accumulation of hypophosphorylated pRb were observed for both the secreted and nonsecreted IFN, suggesting that the nonsecreted IFN may act through a similar pathway. Hep3B cells, an AFP-positive line derived from a patient with hepatocellular carcinoma (HCC), were injected subcutaneously (s.c.) into athymic nude mice to generate established tumors. Intratumoral injection of recombinant adenoviruses expressing secreted as well as the nonsecreted IFN caused suppression of tumor growth. As the AFP promoter is activated in many HCC cells but is silent in normal cells, these constructs may be useful in restricting IFN effects to the tumor cells while reducing toxicity to the neighboring tissues. PMID- 11440638 TI - Porcine caspase-3: its cloning and activity during apoptosis of PK15 cells induced by porcine Fas ligand. AB - We cloned and sequenced cDNA that contained the coding sequence of porcine caspase-3. The open reading frame (ORF) of porcine caspase-3 cDNA was 834 base pairs (bp) in length and encoded 277 amino acids. The predicted amino acid sequence was 88.4%, 86.6%, and 87.7% homologous to the predicted human, murine, and rat amino acid sequences, respectively. The activity of caspase-3 in porcine renal tubular cell line PK15 after recombinant porcine Fas ligand (FasL) stimulation was examined. The enzymatic activity of caspase-3, but not that of caspase-1, was significantly increased after FasL treatment. Western blot analysis also showed that the processing of caspase-3 from proenzyme to mature subunits occurred after FasL treatment. The inhibition of caspase-3 by its specific inhibitor partially prevented the apoptotic cell death of PK15 cells caused by FasL. The porcine caspase-3 cDNA isolated in this study will be useful for the study of apoptotic cell death in pigs and will lead to the discovery of therapeutic uses of caspases and their inhibitors in the prevention of viral and bacterial diseases and tissue injury associated with xenotransplantation and allotransplantation. PMID- 11440639 TI - Rapid changes in hepatitis C virus quasispecies produced by a single dose of IFN alpha in chronically infected patients. AB - The effects of a single dose of 3 international megaunits of interferon-alpha2b (IFN-alpha2b) on hepatitis C virus (HCV) load and quasispecies were examined 24 h after administration in 12 previously untreated, chronically infected patients. All patients had viremia loads appreciably reduced relative to baseline values, thus confirming that the viral load is rapidly affected by IFN-alpha2b. Five patients also exhibited changes in plasma HCV quasispecies composition that were clearly evident by single-strand conformation polymorphism, analysis, thus indicating that one dose of IFN-alpha2b may suffice to produce rapid perturbations in the genetic heterogeneity of circulating HCV. Prior to IFN alpha2b administration, 3 patients exhibited viral quasispecies differences between plasma and peripheral blood mononuclear cells (PBMC). Interestingly, in 2 such patients, the viral quasispecies found in the 24-h plasma resembled that in the pre-IFN PBMC. The latter finding raises the possibility that in these patients, the differences in quasispecies composition between pre-IFN and post IFN plasma resulted from increased representation of lymphoid tissue-originated variants in the latter sample, possibly because of poor sensitivity to IFN alpha2b of HCV replication in the lymphoid compartment. PMID- 11440640 TI - Combined sequential treatment with interferon and dsRNA abrogates virus resistance to interferon action. AB - Many viruses have evolved mechanisms to resist the action of interferon (IFN). These include production of viral gene products that sequester double-stranded RNA (dsRNA) and of small helical RNA. These potentially prevent activation of dsRNA-dependent pathways of IFN action or block expression of cellular genes activated exclusively by dsRNA that may contribute to the antiviral state. Thus, dsRNA might be rate limiting in the development of an IFN-mediated antiviral state. In support of this hypothesis, dsRNA added exogenously to IFN-treated cells in the form of poly(rI):poly(rC) is shown to establish in a dose-dependent manner an antiviral state against two viruses otherwise highly refractory to IFN action, avian reovirus (ARV) and Newcaste disease virus (NDV). Cells exposed singly to high doses of IFN or dsRNA reduced the plaque-forming capacity of these viruses on chicken embryo cells 2-fold. When used in combination, there was up to a 100-fold reduction. In order to abrogate IFN resistance, dsRNA must be added after, not before, an IFN-mediated latent antiviral state is established. dsRNA added exogenously is thought to achieve the threshold required for activation of dsRNA-dependent pathways of IFN action or to induce some dsRNA-stimulated gene whose product acts synergistically with that of some IFN-stimulated gene. The combined sequential treatment with IFN and dsRNA may be useful in overcoming the anti-IFN activity of viruses of clinical interest or in other clinical conditions. PMID- 11440641 TI - Human primary CD4 + T cells activated in the presence of IFN-alpha 2b express functional indoleamine 2,3-dioxygenase. AB - Indoleamine 2,3-dioxygenase (IDO) is the rate-limiting enzyme in the catabolism of tryptophan. By creating a local microenvironment in which levels of tryptophan are low, IDO-expressing antigen-presenting cells (APC) could regulate T cell activation. This may be relevant to control both viral and bacterial replication as well as neoplastic cell growth. Interferon-alpha (IFN-alpha) is an antiviral cytokine affecting cellular differentiation. In addition, it reduces proliferation of CD4(+) T cells by several molecular mechanisms. To dissect the molecular steps responsible for the INF-mediated antiproliferative activity, we sought to determine whether activated primary CD4(+) T cells in the presence of IFN-alpha would produce IDO. We demonstrate here that IDO mRNA is not present in resting CD4(+) T cells. Stimulation with anti-CD3 plus interleukin-2 (IL-2) induces expression of IDO mRNA (about 2000 copies/150,000 cells), as determined by semiquantitative RT-PCR. When cells were stimulated in the presence of IFN alpha, expression of IDO mRNA was significantly increased (more than 12,000 copies/150,000 cells). Functional analysis of IDO activity paralleled the results obtained with RT-PCR, demonstrating increased production of active enzyme in CD4(+) T cells stimulated in the presence of IFN-alpha. Our results indicate that IFN-alpha modulates levels of IDO produced by activated CD4(+) T cells. This would likely affect bystander cells by modifying levels of tryptophan in the local microenvironment. PMID- 11440642 TI - Lipopolysaccharide of Haemophilus influenzae induces interleukin-5 mRNA expression in human peripheral blood mononuclear cells. AB - Haemophilus influenzae is the bacterial species most often isolated from sputum of patients with chronic obstructive pulmonary disease (COPD). In this study, we examined the induction of interleukin-5 (IL-5) mRNA expression in human peripheral blood mononuclear cells (PBMC) stimulated with lipopolysaccharide (LPS) from H. influenzae to try to predict the effect of H. influenzae infection on the eosinophilic inflammation in COPD. Detection of IL-5 mRNA by RT-PCR showed that LPS from H. influenzae induced IL-5 mRNA expression in PBMC at a concentration of 1 microg/ml. Furthermore, the level of expression of IL-5 mRNA induced by LPS correlated with the amount of IL-5 protein in the culture supernatant. Inhibition of LPS-induced IL-5 mRNA expression by anti-CD14 antibody and diminution of this in a CD3(+) -cell-depleted fraction of PBMC, respectively, suggested that CD14 molecules were required for the increase in IL-5 mRNA and that T lymphocytes were the principal source of IL-5 mRNA expression in PBMC. Briefly, the IL-5 mRNA expression induced by LPS would be based on LPS-activated monocytes interacting with T lymphocytes to produce IL-5. These results may explain the role that colonization with H. influenzae plays in eosinophilic inflammation in patients with COPD. PMID- 11440643 TI - Signal transducers and activators of transcription (Stat) are detectable in mouse trigeminal ganglion neurons. AB - We studied signal transducers and activators of transcription (Stat) expression in mouse trigeminal ganglia (TG) to gain an understanding of herpes simplex virus (HSV) infection and reactivation. Mouse TG were harvested and were either frozen for Western blot analysis or preserved in 4% paraformaldehyde for subsequent immunohistochemistry study. The thawed specimens were homogenized, and nuclear/cytoplasmic extractions were performed for Western blots and immunoprecipitation. Immunohistochemistry showed that Stat1, Stat3, Stat4, Stat5b, and phosphotyrosine Stat3 localized to TG neurons, not surrounding satellite cells. Western blot of TG nuclear and cytoplasmic extracts confirmed the presence of these Stat at the appropriate molecular weights. Stat2 was undetectable in TG by these methods. Immunoprecipitation of TG nuclear extracts did not confirm the presence of Stat-Stat dimers in these specimens. These studies show that several Stat, including phosphotyrosine Stat3, are present in TG neurons, the site of HSV latency, where they could act upon latent viral DNA to effect reactivation. PMID- 11440644 TI - [Immigrant patients in primary care. Are we prepared?]. PMID- 11440645 TI - [Self-perception of health in non-institutionalised elderly]. AB - OBJECTIVE: To evaluate the self-perception of health (SH) of non institutionalised over-65s. To relate this SH to age, sex, social and economic level, and use of health services. DESIGN: Descriptive cross-sectional study on a sample stratified by age and sex. SETTING: City of Avila.Participants. 425 interviews representative of the census of the over-65s, using the SF-36 Health Questionnaire and a complementary questionnaire to gather information on social and economic levels and use of health services. MAIN RESULTS: The QWB well-being index was at 0.67 (0.69 men; 0.65 women), dropping with age (0.70 in < 70s; 0.59 in > 84 years old). The highest scores on the questionnaire were in the dimensions social function (81.1) and emotional role (79.8); and the worst in general health (51.9) and vitality (58.5), with men always having higher scores (p < 0.05, except in physical function [p = 0.05] and general health [p = 0.47]). At greater ages the scores were worse, with physical function (-50.6%) and physical role (-45.7%) more affected by age, and mental health (-12.1%), general health (-16.1%) and emotional role (-17.8%) less affected. Men perceived their health as more stable than women did (63.7% vs. 44.6%; p < 0.001). There was self perception of better health, the higher the social and economic levels (p < 0.05 in all dimensions), the greater the educational level reached (p < 0.05 except in social function) and the less the health services were used (p < 0.05, except for mental health-hospital admission). CONCLUSIONS: The Index of Well-being of our elderly people is comparable to that recorded in other samples. Men had self perception of better health than women and saw their health as more stable. SH gets significantly worse with age. Higher economic and social levels and less use of health services are related to better SH. PMID- 11440646 TI - [Communication, diagnosis and treatment of urinary incontinence in the elderly in a basic health area]. AB - OBJECTIVE: To investigate the frequency in which aged people who suffers on urinary incontinence (UI) talk about this problem with primary care professionals, variables related to no healthcare-seeking as well as diagnostic and therapeutic habits of these professionals. DESIGN: Cross-sectional study.Location. Basic Health Zone of Cabra (Cordoba, Spain). PATIENTS: A random age-stratified sample of 869 patients selected from a total of 5139 persons >= 65 years of age.Interventions. Home interview. MEASUREMENTS: Proper questionnaire with questions about reporting of incontinence to health professionals and the procedures used for routine screening as well as diagnostic and therapeutic assessment of incontinence. A logistic regression model was elaborated to identify explanatory factors for reporting incontinence to healthcare professionals. RESULTS: Of 330 incontinents, only 32% reported their problem to healthcare professionals. The remaining 68% did not report their incontinence because most thought it was a minor problem and/or part of the ageing process. The explanatory factors for seeking medical help were the frequency, duration and impact of incontinence, and dependence in self-care. Physicians performed routine screening for incontinence in 10% of 827 inquired aged. In 1 out of 5 incontinents, the physician did not take any diagnostic or therapeutic measures. CONCLUSIONS: Seeking of UI medical help was very limited and depends on variables related to perception in symptom's changes. Diagnosis and treatment done by professionals was suboptimal. Severity of UI conditions diagnosis and treatment. PMID- 11440647 TI - [First experiences in evaluating the clinical competence of family doctors in Catalonia]. AB - OBJECTIVES: To construct a valid instrument to evaluate the clinical competence of family doctors and establish professional standards. DESIGN: Construction of an objective and structured clinical evaluation test by the Test Committee of the Catalan Society of Family and Community Medicine (SCMFiC) with 27 representative clinical situations of primary care practice. This used various evaluative instruments and lasted 5 hours and 30 minutes. SETTING: 2 Barcelona health districts for the two runnings of the test. PARTICIPANTS: Family doctors, SCMFiC members, who took part voluntarily after having been invited by letter. MEASUREMENTS AND MAIN RESULTS: 23 doctors took part in the first running of the test. Their average age was 36.6 (SD 6.9) and 52.2% of them were women. 46 doctors took part in the second test. Their average age was 30.9 (SD 2.1) and 76.1 were women. The test had an overall reliability coefficient (Cronbach's alpha) of 0.83 the first time and 0.65 the second time. The overall mean score of the participants the first time was 56.9 (SD 19.2), with 100 the top score possible, and 62.1 (SD 4.1) the second time. CONCLUSIONS: This SCMFiC experiment is the first time the clinical competence of family doctors has been assessed in our area. It was a reliable and valid test to assess the competence of our health professionals. PMID- 11440648 TI - [What lines of action should the Valencia Society of Family and Community Medicine adopt over the next 4 years?]. AB - OBJECTIVE: To identify the areas of activity in which the Valencia Society of Family and Community Medicine (SVMFyC) should become involved over the next four years. DESIGN: Qualitative research.Setting. SVMFyC members. PATIENTS AND OTHER PARTICIPANTS: 27 experts belonging to the SVMFyC took part. INTERVENTIONS: Qualitative consensus-seeking techniques. Reliability and validity of the technique were ensured through triangulation and the selection of experts from among the different professional groups within the SVMFyC. MEASUREMENTS AND MAIN RESULTS: To determine the recommended lines of action, productivity, spontaneous representativity, intensity of recommendation and degree of agreement were analysed. The priority lines recommended were: defending the MIR path, proposing reforms in undergraduate study plans, watching over the transparency of job selection procedures and the annual OPE selection, promoting the professional degree course, creation of posts for teachers in family and community medicine, defining the size of the population registered with a doctor, proposing an incentives list and studying alternatives to uniform salaries in the form of target-linked remuneration. CONCLUSIONS: The lines of action recommended by the experts were established. PMID- 11440649 TI - [Study of mortality in a urban basic health area]. AB - OBJECTIVE: To find the pattern of causes of mortality in the Chantrea (Navarra) Health District and the trends in main causes of death between 1987 and 1995. DESIGN: Descriptive study. SETTING: Urban primary care centre. STUDY POPULATION: Population belonging to the Chantrea Health District.Interventions. 1986 data from the Municipal Roll of inhabitants of Chantrea were crossed with the Navarra mortality figures to find the deaths occurred between 1986 and 1995. Overall mortality and mortality for the main reasons were analysed for both sexes and for the periods 1987-1990 and 1991-1995. RESULTS: Between 1991 and 1995, Chantrea had a mortality rate close to that of the European population: 711 per 100000 inhabitants for men and 403 per 100000 for women. The main causes of death were circulatory diseases, tumours, respiratory diseases and external causes. Overall mortality rates dropped by 11.8% among men and by 8% among women from the 1987 1990 period to the 1991-1995 period. Statistically significant drops in mortality were noted for circulatory diseases and for malignant oesophageal tumours in men. AIDS as cause of death increased significantly among men. CONCLUSIONS: Low rates of overall mortality in men and women of Chantrea and, in particular, mortality rates for circulatory disease were below the figures recorded in most European countries. Other causes of death, such as traffic accidents, AIDS or malignant tobacco-related tumours had moderately high rates. PMID- 11440650 TI - [Failure of rapid serology for Helicobacter pylori as diagnostic method in primary care consultation]. AB - OBJECTIVE: To evaluate the validity of a rapid serology test for diagnosing Helicobacter pylori infection in our area. DESIGN: Longitudinal, prospective study. SETTING: Urban primary care centre. MEASUREMENTS: As reference tests for defining the presence/absence of Helicobacter pylori, histology and the urease gastric mucous membrane test (Jatrox(R) test) were used. Both tests can be either positive or negative, without intermediate values. RESULTS: 47 consecutive patients were included. Histology (confirmed with the urease test) showed infection in 38 cases (80.85% prevalence). The results of rapid serology (95% CI) were: sensitivity 0.66 (0.51-0.81), specificity 1 (1-1), positive predictive value 1 (1-1), and negative predictive value 0.41 (0.2-0-61). CONCLUSIONS: In a population with high prevalence of infection, a positive in the rapid serology test confirms infection. However, a negative does not discount infection, which significantly reduces the test's overall diagnostic value. This confirms in the primary care context the findings of hospital-based studies. PMID- 11440651 TI - [The Ottawa ankle guidelines: analysis of their validity as clinical decision guidelines in the indication of X-rays for ankle and/or middle-foot injuries]. AB - OBJECTIVE: To analyse the validity of the Ottawa ankle guidelines (OAG) as clinical decision guidelines in the indications of X-rays for ankle and/or middle foot traumas in primary care. DESIGN: Observational, with application of the OAG and prospective measurement of the results.Setting. Hospital casualty. PATIENTS: Adults who attended casualty for ankle or middle-foot traumas between 1st June 1999 and 31th March 2000. Criteria for exclusion were: under 18, pregnancy, grave sensory and/or awareness disturbances, multi-trauma or multi-contusion patients, traumas over a week old, skin lesions as side-effects of the trauma, X-ray in other department, high inflammation or oedema hindering palpation of bone protuberances. MEASUREMENTS: Application of the OAG and X-ray on all patients, regardless of the result of the OAG. Calculation of sensitivity, negative predictive value, specificity and positive predictive value. RESULTS: 56 of a sample of 494 patients had a fracture (11.34%), 34 in the malleolus area (6.9%) and 22 in the middle-foot area (4.44%). OAG sensitivity was 96.43% (95% CI, 94.8 98). Negative predictive value was 97.22% (95.77-98.67). Specificity was 15.98% (12.75-19.21), and positive predictive value was 12.8% (9.86-15.74). CONCLUSIONS: The OAG are valid in primary care as guidelines to decide whether to request X rays for patients with ankle or middle-foot traumas. PMID- 11440652 TI - [Asthma and COPD in Internet]. PMID- 11440653 TI - [Wellbeing or happiness medications?]. PMID- 11440654 TI - [Unexplained fever in patient with Down's syndrome]. PMID- 11440655 TI - [Smoking in young people. Interdisciplinary approach]. PMID- 11440656 TI - [Secondary arterial hypertension in asymptomatic young man]. PMID- 11440657 TI - [Hypothyroidism and sleep apnea syndrome. Report of a case]. PMID- 11440658 TI - [Prevalence of caries in 6-12 year olds in the health area of Burela (Lugo)]. PMID- 11440660 TI - [Osteomyelitis: An unresolved matter for the 21st century]. PMID- 11440661 TI - [Antifungal susceptibility of emerging yeast pathogens]. AB - BACKGROUND: To study the antifungal susceptibility of emerging yeast pathogens to know their possible resistance under the need of applying a treatment. MATERIAL AND METHODS: We investigated the in vitro susceptibility of 69 yeast strains isolates of clinical samples, belonging to 24 different species, to amphotericin B, fluconazole, itraconazole, ketoconazole and 5-fluorocytosine. RESULTS: Only 9 species showed susceptibility to all antifungal agents: Candida famata, C. guillermondii, C. holmii, C. kefyr, C. pelliculosa, C. rugosa, C. utilis, C. zeylanoides y Trichosporon cutaneum; the rest of them presented resistance to some antifungal agent. C. haemulonii, Pichia farinosa and Trichosporon mucoides were resistant to amphotericin B; C. haemulonii, C. inconspicua, C. lusitaniae, C. norvegensis, C. pintolepesii, C. valida, P. ohmeri, Rhodotorula glutinis, R. minuta, R. mucilaginosa and Saccharomyces cerevisiae were resistant to azoles; Blastoschizomyces capitatus and C. lipolytica were resistant to 5-fluorocytosine. CONCLUSIONS: The resistance of emerging yeast pathogens to amphotericin B and 5 fluorocytosine is low, while resistance to azoles is significative, especially to fluconazole (36%). Many of this yeasts present problems of intrinsic resistance. In yeast infections, the correct identification of species and the study of the in vitro susceptibility is important in order to choose the most adequate antifungal treatment. PMID- 11440662 TI - [Abscess of the psoas muscle. Description of a series of 23 cases]. AB - BACKGROUND: Abscess of the psoas muscle (PA) is an infrequent disease of difficult diagnosis. During the last decade, the number of cases has increased because of the raising use of radiology tecniques: ecography, computerized tomography and magnetic resonance nuclear. METHODS: The presentation and management of psoas abscess was studied retrospectively in 23 patients from 1992 2000. RESULTS: Sixteen of 23 abscesses were regarded as secundary: spondylodiscitis and pyelonefritis were most frequent pathologic processes. Homolateral pain in the flank area and hip were the usual manifestations. The duration of symptoms prior to the diagnosis was superior than 7 days. Staphylococcus aureus was the most common pathogen, followed by Escherichia coli and Mycobacterium tuberculosis. All abscesses were diagnosed by computerized tomography images. Seven patients underwent percutaneous drainage, while nine received surgical debridement. Four of the patients with psoas abscess died and only three relapsed. CONCLUSIONS: Not specific symptoms and signs and subacute presentation difficult diagnosis of AP. High range antibiotics and drainage (percutaneous or surgical) should be considered as the election treatment. PMID- 11440663 TI - [Genital infection and infertility]. AB - BACKGROUND: Genital infection is the most important cause of infertility worldwide, affecting not only Fallopian tubes but all anatomic urogenital sites, both male and female. METHODS: We present a randomized,prospective and normalized study about sexually transmitted diseases(STD). We include 487 patients, 376 of whom were infertile and the remaining 111 were not and act as control group. RESULTS: 47.3% of infertile patients showed at least one infection: 10.7% had Chlamydia trachomatis infection, whereas only 0.3% had gonococal infection. We found none syphilis. 12.9% of the patients showed yeast belonging to genus Candida, 5% bacterial vaginosis, 3.8% Escherichia coli and 0.3% Klebsiella pneumoniae. The percentage of isolation of Ureaplasma urealyticum and Mycoplasma hominis were 23.5% and 4.8%, respectively. We detected antibodies against Hepatitis B (any serological marker) in 7.8% of the cases. Chlamydial infection and the presence of U. urealyticum were related with infertility (X 12=6,070, p<0.005 and X12=8,782, p<0.005,respectively). CONCLUSIONS: We think these results conclude that is necessary to perform routine tests to screen for C. trachomatis, N. gonorrhoeae, and for infections caused by micoplasma as well, among infertile patients. These patients must be considered of being at risk of acquiring STD, since the percentage of these diseases is higher than the percentage we found among the control group. It is important to insist on prevention and early diagnosis and treatment as a main goal to decrease the number of tubal occlusion and infertile couples. PMID- 11440664 TI - [Detection of Brucella melitensis by BACTEC 9050 system]. AB - OBJECTIVES: To evaluate BACTEC 9050 system capacity for detection of bacteremia due to Brucella spp. MATERIAL AND METHODS: 16 febrile patients were studied during an epidemic infection for Brucella spp. Suspicious of brucellosis was serologically confirmed with Rose Bengal test and agglutination tube test. Only one blood culture was processed per patient. Ten millilitres of blood were inoculated in a Bactec Plus aerobic/F bottle and incubated in BACTEC 9050 automatic system. The bottles were kept in incubation during 21 days, and they were subcultured when the machine detected its growth; if not, a blind subculture was performed after 21 days. RESULTS: 13 of 16 patients showed bacteriemia (81.2%): 11 patients were detected by BACTEC 9050 system and 2 patients by blind subculture after 21 days. A positive result appeared in 149.8 hours (6.2 days) as a mean. Earlier detections were seen in 83 hours (3.4 days) and the latest ones at 245 hours (10.2 days). The aetiology agent of the epidemic infection was Brucella mellitensis biotype 1. We found significative differences comparing the BACTEC 9050 with BACTEC 9120/9240 systems (p<0.03). CONCLUSIONS: Incubation protocols of 5 days are not useful for BACTEC 9050 system in the case brucellosis suspicious. Protocols of 7 would detect only 69.2% of the cases. PMID- 11440665 TI - [Malaria in non-endemic countries. Review of the current situation]. PMID- 11440666 TI - [Recurrent acute bacterial meningitis]. PMID- 11440667 TI - [Parasitologic finding in a Saharan girl]. PMID- 11440668 TI - [Cough and bloody expectoration a patient with antecedent tuberculosis]. PMID- 11440669 TI - [Septic arthritis of the elbow due to Actinomyces spp. and a foreign body]. PMID- 11440670 TI - [Prevotella denticola endocarditis in an intravenous drug abuser]. PMID- 11440671 TI - [Yeast growing time in continuously monitored blood cultures]. PMID- 11440672 TI - [Bacteremia due to Providencia rettgeri]. PMID- 11440673 TI - [Reversible dementia from an infectious cause: the importance of the Argyll Robertson pupil in the diagnosis of neurosyphilis]. PMID- 11440674 TI - [Corynebacterium riegelii urinary tract infection]. PMID- 11440675 TI - [Tenosynovitis caused by Cryptococcus neoformans in a patient with AIDS]. PMID- 11440676 TI - [Human immunodeficiency virus/hepatitis C virus co-infection: do we have the right not to treat our patients?]. PMID- 11440677 TI - [Antiretroviral drug resistance]. PMID- 11440679 TI - [Anti-GQ1b antibodies: usefulness of its detection for the diagnosis of Miller Fisher syndrome]. AB - BACKGROUND: To study the presence of anti-GQ1b antibodies as a tool for the diagnosis of Miller-Fisher syndrome (MFS). PATIENTS AND METHOD: We studied 54 patients with probable diagnosis of MFS and 10 patients diagnosed as Guillain Barre syndrome plus ophthalmoplegia (1 case), Bickerstaff's encephalitis (1 case), relapsing ophthalmoplegia (7 cases) and relapsing diplopia (1 case). Results were compared with 130 patients with other disimmune neuropathies. Antibodies were detected by ELISA and checked by thin layer chromatography. Campylobacter jejuni serology was studied using a complement fixation test. RESULTS: Diagnosis of MFS was confirmed in 38 patients. A 97.3% were positive for GQ1b, being all negative for Campylobacter jejuni serology. A second test after 4 5 weeks of nadir was negative in 84.2% (16/19), concomitant with clinical recovery. CONCLUSIONS: Anti-GQ1b antibodies are useful markers for the differential diagnosis of MFS, specially with some acute brainstem disorders. Testing must be performed during the first four weeks of clinical course. This correlation between the triad ataxia, arreflexia and ophthalmoplegia and anti GQ1b antibodies confirms that they are highly specific of MFS. PMID- 11440680 TI - [Risk factors for nosocomial sepsis: a case-control study]. AB - BACKGROUND: The objective of this study was to identify the risk factors associated with the nosocomial sepsis syndrome according to the criteria of the American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference. PATIENTS AND METHOD: A 1-year prospective case-control study matched for sex, age (+/- 5 years), and pre-infection hospital stay (+/- 1 day) was performed in a 1,200-bed university hospital. Cases were selected according to the above criteria. Controls were randomly selected from the daily list of hospitalized patients. Crude and adjusted odds ratios (OR) were determined. RESULTS: 346 cases and 346 controls were included. Multivariate analysis identified the following intrinsic risk factors: coma in the 48 hours before sepsis (OR: 15.1; CI 95%, 5.6-41.2), renal failure (OR: 3.4; CI 95%; 1.5-10.8), neoplasm (OR: 2.4; CI 95%, 1.1-5.1), prosthesis material (OR: 2.7; CI 95%, 1.0 7.8), and serum albumin concentration at admission lower than 3.1 g/dl (OR: 5.3; CI 95%, 2.3-12.4). Main extrinsic risk factors were: previous nosocomial infection (OR: 12.5; CI 95%, 1.61-96.3), intensive care unit (ICU) stay (OR: 10.6; CI 95%, 3.1-36.2), naso-gastric tube (OR: 8.4; CI 95%, 2.3-31.3), indwelling urinary catheter (OR: 5.0; CI 95%, 1.4-18.9), H2 blockers treatment (OR: 5.0; CI 95%, 1.6-15.2), and IV central line (OR: 4.1; CI 95%, 1.2-14.0). CONCLUSION: In our study, main risk factors for development of nosocomial sepsis were presence of coma in the 48 hours before sepsis, ICU stay, and prior cross infection during hospitalization. PMID- 11440681 TI - [Prevalence of tuberculosis in a population from Kosovo sheltered in Catalonia, Spain]. AB - BACKGROUND: Prevalence of tubeculosis (TB) among refugees is high. The goal was to detect TB active cases in a group of refugees from Kosovo and to describe the prevalence of TB infection. PATIENTS AND METHOD: 222 people sheltered in Catalonia, Spain, were tested for TB using the Mantoux method and a chest x-ray. The odds ratio (OR) was adjusted by the age of presenting a positive PPD test for different diameters of induration according to the previous BCG vaccination history. RESULTS: No cases of active TB were detected and 82% of the population was found to be vaccinated with BCG. The OR of presenting a positive PPD was higher, and statistically significant, in vaccinated subjects compared to non vaccinated ones for indurations >= 5 mm and >= 10 mm (OR: 4.5 and 3.6 respectively). CONCLUSIONS: BCG-vaccinated subjects from this population can be considered to be infected by Mycobacterium tuberculosis as long as the induration is higher than 15 mm. PMID- 11440682 TI - [Uranium and disease]. PMID- 11440683 TI - [Management of chronic hepatitis C infection in patients infected with the human immunodeficiency virus: from what we know to what we ignore]. PMID- 11440684 TI - [Human resources in the health care system: pending reforms]. PMID- 11440685 TI - [Severe sepsis and septic shock: crossroad of inflammation and coagulation]. PMID- 11440686 TI - [Measurements of quality of life related with health. Basic concepts and cultural adaptation]. PMID- 11440687 TI - [Chronic myelomonocytic leukemia presenting as Guillain- Barre syndrome]. PMID- 11440688 TI - [Long term survival in a patient with aggressive Rosai-Dorfman disease treated with interferon alpha]. PMID- 11440689 TI - [Absence of Candida dubliniensis in a population of institutionalized elderly patients]. PMID- 11440690 TI - [Functional decline in nonagenarians admitted at acute care hospitals]. PMID- 11440692 TI - [Frequency and risk of bronchopulmonary neoplasia related to asbestos]. AB - BACKGROUND: The goal of this study was to determine the prevalence of asbestos related lung cancer and the importance of the occupational exposure to this inorganic fibre as a risk factor. PATIENTS AND METHODS: We performed a cross sectional study of 82 patients with lung cancer (mean age 62 SD 9 years) and 53 patients without pleuropulmonary disease (63 SD 13 years). The occupational exposure to asbestos was determined by a questionnaire. We determined the concentration of asbestos bodies (AB) in bronchoalveolar lavage (BAL) (93 patients) or lung tissue (42 patients) after chemical digestion, with the results being expressed as AB/mL BAL or AB/g dry lung, respectively. A concentration higher than 1 AB/mL or 1,000 AB/g was considered as marker of high asbestos burden in lung tissue, which could be potentially responsible for pleuropulmonary disease. The importance of asbestos occupational exposure as a risk factor for lung cancer was determined using logistic regression models. RESULTS: 25 patients with lung cancer reported occupational exposure to asbestos (30%) and in 13 out of them AB were detected in BAL or lung tissue (24%), at high concentrations in 3 cases (4%). Six patients from the group without pleuropulmonary disease reported occupational exposure to asbestos (11%) and in 13 out of them AB were found in some samples (24%), with no case having high concentrations. In the univariate logistic regression analysis, diagnosis of bronchial neoplasia was associated with both smoking (OR 10.10, 95% CI 3.50-29.13) and occupational exposure to asbestos (OR 3.69, 95% CI 1.39-9.77). The association between asbestos exposure and lung cancer persisted statistically significant after adjustment for smoking (OR 2.80, 95% CI 1.00-7.84). CONCLUSION: In Spain, lung cancer was related to occupational exposure to asbestos in 4% of cases, and it appeared to exist a synergistic effect of smoking. Occupational exposure to this inorganic fibre doubles the risk of suffering from lung cancer. PMID- 11440693 TI - [Impact in the quality of health care and cost-effectiveness analysis of the reform of an emergency medicine service]. AB - BACKGROUND: To determine the effects that the reform of an emergency department (ED) have on efficacy, health care quality and efficiency. MATERIAL AND METHOD: Reforms consisted of 50% and 34% increases in structural and human resources, respectively. Roles of each ED member were redetermined, new assistance pathways were implemented, and the relationship between ED and the remaining hospital departments was reassessed. As efficacy markers, we determined the number of patients waiting to be attended (Pesp), the waiting time of patients to receive medical assistance (T(esp)),and the total waiting time of patients staying in the ED (T(total)). As health-care quality markers, we determined the percentage of patients leaving ED without having been visited by a physician(IPNV), the percentage of patients who were visited again (IPR),and the percentage of mortality (IPF). We also quantified the number of visits to the ED. All these data were obtained daily over 3 weeks, before (February 1999) and after (February 2000)the reforms. Effectiveness was estimated from the ratio P(total)/T(esp)(E1) and P(total)/P(esp) (E2). Costs were recorded for both periods and a cost effectiveness analysis was performed to study the efficiency. RESULTS: In 2000, the number of visits increased by +12% (CI 95%: 2% to 22%). Despite this increase, we observed an improvement of most efficacy and health-care quality markers after the ED reforms. E1 increased by 996% (CI 95%: 335% to 1,658%) and E2 increased by 186% (CI 95%: -23%to 395%). Cost-effectiveness analysis showed 70% (CI 95%: 33%to 107%) and 56% (CI 95%: 18% to 94%) increases regarding E1 and E2, respectively, after the reforms. CONCLUSIONS: Providing ED with the necessary resources leads to an objective improvement of its efficacy and health-care quality and, consequently, the service and quality perceived by users improve. Despite the total cost increase after the ED reforms, efficiency also improves. PMID- 11440694 TI - [Malposition of central venous catheters in hospitalized patients]. AB - BACKGROUND: At the time of inserting a central catheter, it is recommended to check that its distal tip is positioned outside the heart cavities. Our objective was to determine the degree of fulfillment of this recommendation. PATIENTS AND METHOD: Cross-sectional study on patients carrying a central venous catheter who were admitted to conventional areas of a university hospital. We considered as "risk catheter" any catheter whose distal tip was located within the cardiac cavities. RESULTS: 34 out of 362 hospitalized patients had a central venous catheter. Fifty per cent of them were found to have a "risk catheter". In spite of its X-ray confirmation, no patient underwent a subsequent modification of the distal tip position. CONCLUSIONS: We have noticed a high percentage of patients with central venous catheters whose insertion supposed a cardiovascular risk. Unawareness of the serious complications that may arise from this fact probably justifies a high prevalence of misplacements. PMID- 11440695 TI - [Advance directives document]. PMID- 11440696 TI - [Bioethics for clinicians: a need, an initiative, a proposal]. PMID- 11440697 TI - [Moral deliberation: the method of clinical ethics]. PMID- 11440698 TI - [40 years ago in Medicina Clinica. The introduction of melphalan in Spain]. PMID- 11440699 TI - [Drinking water chlorination and adverse health effects: review of epidemiological studies]. PMID- 11440700 TI - [Symptomatic hyperlactemia in patients infected with the human immunodeficiency virus treated with nucleoside analogs]. PMID- 11440701 TI - [Focal pachymeningitis as form of presentation of Wegener's granulomatosis]. PMID- 11440702 TI - [Magnetic resonance cholangiography in the diagnosis of massive pleural effusion secondary to a pancreatico-pleural fistula]. PMID- 11440703 TI - [Prevalence of osteoporosis assessed by densitometry in the Spanish female population]. PMID- 11440705 TI - [Hemoperitoneum secondary to intrahepatic pseudoaneurysm as a complication of thoracocentesis]. PMID- 11440707 TI - [Important changes, decisive moments]. PMID- 11440708 TI - [Sentinel lymph nodes in breast cancer. Multidisciplinary approach; critical analysis of the learning curve; results obtained with a radiopharmaceutical not previously utilized]. AB - Detection of the sentinel node is slowly becoming a new and promising procedure to stage patients suffering from breast cancer. Various articles have attempted to analyze objectively the predictive value of the histopathological examination to reveal the presence of metastatic cells in the axillary node without having to carry out an axillary dissection. In this paper, 60 patients with breast cancer (T up to 3 cm, clinically negative axilla) have been studied. A colloid bound to 99m Technetium was injected. Intra-surgical radioguided detection of the sentinel lymph node was performed and this was excised and its pathology was analyzed. After a complete axillary dissection was performed. With this technique it was possible to predict the condition ot the axilla in 96.5% of the cases and the sensitivity of the method was 92%. A training program on this technique has been developed and its application is suggested. The results of the use of a radiopharmaceutical which has not been previously documented in the published literature is discussed. PMID- 11440709 TI - Diagnostic value of 99mTc MIBI scintimammography in patients with breast lesions. AB - This study has aimed to determine the diagnostic value of the scintimammography with 99mTc-MIBI in the detection of primary breast cancer and to verify its clinical usefulness. Sixty-three female patients (age range 27-83) with breast lesions detected by physical examination, ultrasonography, and/or mammography were prospectively included in this study. An anterior and prone lateral planar 99mTc-MIBI scintimammography was performed on all patients. The final diagnosis was achieved by histopathological examination in 49 patients and clinical followup (at least one year) in 14 patients. Histopathological diagnosis confirmed that 15 lesions were malignant and 34 lesions were benign. All the malignant lesions were larger than 1 cm. The 99mTc-MIBI scintimammography showed increased focal uptake in 17 lesions (two of them were benign and the other 15 were malignant). The diffuse heterogeneous uptake pattern was considered as benign. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy values for breast carcinoma were calculated as 100%, 96%, 88%, 100%, 97%, respectively. We concluded that 99mTc-MIBI scintimammography could be a valuable method in the differentiation of malignant breast neoplasm larger than 1 cm size from benign ones, especially when the uptake pattern was considered. PMID- 11440710 TI - Journey to the center of the cell. PMID- 11440711 TI - Signaling downstream of Eph receptors and ephrin ligands. PMID- 11440712 TI - The pancreatic beta cell heats up: UCP2 and insulin secretion in diabetes. PMID- 11440713 TI - Rosa Beddington. PMID- 11440714 TI - Molecular insights into polyubiquitin chain assembly: crystal structure of the Mms2/Ubc13 heterodimer. AB - While the signaling properties of ubiquitin depend on the topology of polyubiquitin chains, little is known concerning the molecular basis of specificity in chain assembly and recognition. UEV/Ubc complexes have been implicated in the assembly of Lys63-linked polyubiquitin chains that act as a novel signal in postreplicative DNA repair and I kappa B alpha kinase activation. The crystal structure of the Mms2/Ubc13 heterodimer shows the active site of Ubc13 at the intersection of two channels that are potential binding sites for the two substrate ubiquitins. Mutations that destabilize the heterodimer interface confer a marked UV sensitivity, providing direct evidence that the intact heterodimer is necessary for DNA repair. Selective mutations in the channels suggest a molecular model for specificity in the assembly of Lys63 linked polyubiquitin signals. PMID- 11440715 TI - Crystal structure of glycogen synthase kinase 3 beta: structural basis for phosphate-primed substrate specificity and autoinhibition. AB - Glycogen synthase kinase 3 beta (GSK3 beta) plays a key role in insulin and Wnt signaling, phosphorylating downstream targets by default, and becoming inhibited following the extracellular signaling event. The crystal structure of human GSK3 beta shows a catalytically active conformation in the absence of activation segment phosphorylation, with the sulphonate of a buffer molecule bridging the activation-segment and N-terminal domain in the same way as the phosphate group of the activation-segment phospho-Ser/Thr in other kinases. The location of this oxyanion binding site in the substrate binding cleft indicates direct coupling of P+4 phosphate-primed substrate binding and catalytic activation, explains the ability of GSK3 beta to processively hyperphosphorylate substrates with Ser/Thr pentad-repeats, and suggests a mechanism for autoinhibition in which the phosphorylated N terminus binds as a competitive pseudosubstrate with phospho-Ser 9 occupying the P+4 site. PMID- 11440716 TI - Structural basis of the interaction of the pyelonephritic E. coli adhesin to its human kidney receptor. AB - PapG is the adhesin at the tip of the P pilus that mediates attachment of uropathogenic Escherichia coli to the uroepithelium of the human kidney. The human specific allele of PapG binds to globoside (GbO4), which consists of the tetrasaccharide GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc linked to ceramide. Here, we present the crystal structures of a binary complex of the PapG receptor binding domain bound to GbO4 as well as the unbound form of the adhesin. The biological importance of each of the residues involved in binding was investigated by site-directed mutagenesis. These studies provide a molecular snapshot of a host-pathogen interaction that determines the tropism of uropathogenic E. coli for the human kidney and is critical to the pathogenesis of pyelonephritis. PMID- 11440717 TI - Uncoupling protein-2 negatively regulates insulin secretion and is a major link between obesity, beta cell dysfunction, and type 2 diabetes. AB - beta cells sense glucose through its metabolism and the resulting increase in ATP, which subsequently stimulates insulin secretion. Uncoupling protein-2 (UCP2) mediates mitochondrial proton leak, decreasing ATP production. In the present study, we assessed UCP2's role in regulating insulin secretion. UCP2-deficient mice had higher islet ATP levels and increased glucose-stimulated insulin secretion, establishing that UCP2 negatively regulates insulin secretion. Of pathophysiologic significance, UCP2 was markedly upregulated in islets of ob/ob mice, a model of obesity-induced diabetes. Importantly, ob/ob mice lacking UCP2 had restored first-phase insulin secretion, increased serum insulin levels, and greatly decreased levels of glycemia. These results establish UCP2 as a key component of beta cell glucose sensing, and as a critical link between obesity, beta cell dysfunction, and type 2 diabetes. PMID- 11440718 TI - Drosophila fasciclinII is required for the formation of odor memories and for normal sensitivity to alcohol. AB - Drosophila fasciclinII (fasII) mutants perform poorly after olfactory conditioning due to a defect in encoding, stabilizing, or retrieving short-term memories. Performance was rescued by inducing the expression of a normal transgene just before training and immediate testing. Induction after training but before testing failed to rescue performance, showing that Fas II does not have an exclusive role in memory retrieval processes. The stability of odor memories in fasII mutants are indistinguishable from control animals when initial performance is normalized. Like several other mutants deficient in odor learning, fasII mutants exhibit a heightened sensitivity to ethanol vapors. A combination of behavioral and genetic strategies have therefore revealed a role for Fas II in the molecular operations of encoding short-term odor memories and conferring alcohol sensitivity. The preferential expression of Fas II in the axons of mushroom body neurons furthermore suggests that short-term odor memories are formed in these neurites. PMID- 11440719 TI - A role for the segment polarity gene shaggy/GSK-3 in the Drosophila circadian clock. AB - Tissue-specific overexpression of the glycogen synthase kinase-3 (GSK-3) ortholog shaggy (sgg) shortens the period of the Drosophila circadian locomotor activity cycle. The short period phenotype was attributed to premature nuclear translocation of the PERIOD/TIMELESS heterodimer. Reducing SGG/GSK-3 activity lengthens period, demonstrating an intrinsic role for the kinase in circadian rhythmicity. Lowered sgg activity decreased TIMELESS phosphorylation, and it was found that GSK-3 beta specifically phosphorylates TIMELESS in vitro. Overexpression of sgg in vivo converts hypophosphorylated TIMELESS to a hyperphosphorylated protein whose electrophoretic mobility, and light and phosphatase sensitivity, are indistinguishable from the rhythmically produced hyperphosphorylated TIMELESS of wild-type flies. Our results indicate a role for SGG/GSK-3 in TIMELESS phosphorylation and in the regulated nuclear translocation of the PERIOD/TIMELESS heterodimer. PMID- 11440720 TI - Smoothened mutants reveal redundant roles for Shh and Ihh signaling including regulation of L/R asymmetry by the mouse node. AB - Genetic analyses in Drosophila have demonstrated that the multipass membrane protein Smoothened (Smo) is essential for all Hedgehog signaling. We show that Smo acts epistatic to Ptc1 to mediate Shh and Ihh signaling in the early mouse embryo. Smo and Shh/Ihh compound mutants have identical phenotypes: embryos fail to turn, arresting at somite stages with a small, linear heart tube, an open gut and cyclopia. The absence of visible left/right (L/R) asymmetry led us to examine the pathways controlling L/R situs. We present evidence consistent with a model in which Hedgehog signaling within the node is required for activation of Gdf1, and induction of left-side determinants. Further, we demonstrate an absolute requirement for Hedgehog signaling in sclerotomal development and a role in cardiac morphogenesis.[Dedicated to Rosa Beddington, a pioneer in mammalian embryology]. PMID- 11440721 TI - A molecular link between stem cell regulation and floral patterning in Arabidopsis. AB - The homeotic gene AGAMOUS (AG) has dual roles in specifying organ fate and limiting stem cell proliferation in Arabidopsis flowers. We show that the floral identity protein LEAFY (LFY), a transcription factor expressed throughout the flower, cooperates with the homeodomain protein WUSCHEL (WUS) to activate AG in the center of flowers. WUS was previously identified because of its role in maintaining stem cell populations in both shoot and floral meristems. The unsuspected additional role of WUS in regulating floral homeotic gene expression supports the hypothesis that floral patterning uses a general meristem patterning system that was present before flowers evolved. We also show that AG represses WUS at later stages of floral development, thus creating a negative feedback loop that is required for the determinate growth of floral meristems. PMID- 11440722 TI - Termination of stem cell maintenance in Arabidopsis floral meristems by interactions between WUSCHEL and AGAMOUS. AB - Floral meristems and shoot apical meristems (SAMs) are homologous, self maintaining stem cell systems. Unlike SAMs, floral meristems are determinate, and stem cell maintenance is abolished once all floral organs are initiated. To investigate the underlying regulatory mechanisms, we analyzed the interactions between WUSCHEL (WUS), which specifies stem cell identity, and AGAMOUS (AG), which is required for floral determinacy. Our results show that repression of WUS by AG is essential for terminating the floral meristem and that WUS can induce AG expression in developing flowers. Together, this suggests that floral determinacy depends on a negative autoregulatory mechanism involving WUS and AG, which terminates stem cell maintenance. PMID- 11440724 TI - Development of a simple whole blood panel test for detection of human heart-type fatty acid-binding protein. AB - OBJECTIVE: For the diagnosis of acute myocardial infarction (AMI), we have developed a rapid and simple whole blood panel test for the detection of human heart-type fatty acid-binding protein (H-FABP) using one-step immunochromatography. METHODS AND RESULTS: We have developed a whole blood panel test for rapid detection of human H-FABP using a one-step immunochromatography technique. The result of this panel test was not affected by the other contents of the blood such as bilirubin, hemoglobin and others. Furthermore, no cross reactivity of the antibodies was found with other cardiac markers or other tissue type FABPs. The result of this panel test was similar to the diagnostic cut-off value, 6.2 ng of H-FABP per mL of serum which was evaluated by the enzyme-linked immunosorbent assay (ELISA). CONCLUSION: We have developed a simple one-step immunochromatography technique to detect H-FABP in whole blood sample. Further studies are required to identify the value of this point-of-care testing (POCT) as a diagnostic marker for AMI. PMID- 11440725 TI - Oxidative stress indices in IDDM subjects with and without long-term diabetic complications. AB - BACKGROUND: Numerous animal and population studies of diabetes have identified markers of oxidative stress. However, for most markers that have been measured the results are not consistent. In addition, it is less clear whether oxidative stress is related to the development of diabetic complications. The objective of this study was to evaluate a series of plasma markers and leukocyte markers to test the hypothesis that type 1 Insulin Dependent Diabetes Mellitus (IDDM) subjects experience oxidative stress. A related question was whether markers of oxidative stress are higher in IDDM subjects who have developed long-term complications. METHODS: The study population consisted of 22 IDDM subjects with diabetic complications and 22 IDDM subjects without complications, both groups matched by age and gender and with similar HbA1c levels, and 16 nondiabetic control subjects. Plasma levels of organoperoxides were determined by the ferrous oxidation/xylenol orange (FOX) assay, malondialdehyde by the thiobarbituric acid (TBARS) assay, and vitamin E by HPLC. Mononuclear cells and polymorphonuclear cells were analyzed for ascorbic acid by HPLC and for glutathione (GSH) by enzymatic recycling. In addition, GSH peroxidase, GSH transferase and glucose-6 phosphate dehydrogenase levels were determined in both cell fractions. RESULTS: Plasma organoperoxides were significantly elevated in the IDDM subjects compared to controls (p = 0.02) while TBARS and vitamin E levels were not significantly different. In the IDDM subjects, mononuclear cell levels of ascorbic acid were significantly lower (p < 0.02) and levels of GSH were lower, approaching significance (p = 0.07), compared to controls. Ascorbic acid and GSH levels in polymorphonuclear cells were not significantly different between IDDM subjects and controls, nor were enzyme levels different. In addition, the plasma and intracellular indices of oxidative status in IDDM subjects were not different when IDDM subjects with complications were compared to IDDM subjects without complications. CONCLUSION: Demonstration of oxidative stress in IDDM subjects depends upon which markers are measured. This is in agreement with previous studies of oxidative stress in various disease states including diabetes. Plasma levels of organoperoxides may be the most reliable indicators of oxidative stress. However, it is unclear whether elevated plasma organoperoxides indicate a generalized systemic stress or are produced in localized areas. By comparison, oxidative stress indices determined with isolated blood cells may provide a clearer picture. Depressed levels of ascorbic acid and GSH were observed only in mononuclear cells, which are mainly long-lived T lymphocytes. Mononuclear cells antioxidant status may reflect systemic oxidative stress. In this study, neither plasma markers nor intracellular markers of oxidative stress were different in IDDM subjects with long-term diabetic complications compared to subjects without complications. PMID- 11440726 TI - Association between homocysteine and neopterin in healthy subjects measured by a simple HPLC-fluorometric method. AB - OBJECTIVES: Neopterin and homocysteine promote vascular smooth muscle cell proliferation through the activation of nuclear factor(kappa) B. The aim of this study was to investigate the relation between these two compounds in healthy subjects by a rapid HPLC-fluorometric method which simplifies sample pretreatment for the measurement of neopterin in serum. DESIGN AND METHODS: In 40 healthy subjects (45.9 +/- 2.1 yr, mean +/- SEM, 10 males, 30 females) serum neopterin concentrations were measured by HPLC-fluorometry and enzyme-linked immunusorbant assay-ELISA and the results were compared. Urinary neopterin and plasma total homocysteine concentrations were assayed by HPLC-fluorometry. RESULTS: Serum neopterin concentrations measured by HPLC and ELISA were 7.5 +/- 0.4 and 7.4 +/- 0.3 nmol/L, respectively, r = 0.92, p < 0.01. Urinary neopterin level was 163.9 +/- 11.0 nmol/mmol creatinine and plasma total homocysteine 7.6 +/- 0.4 micromol/L. A significant positive correlation was observed between serum neopterin and plasma total homocysteine (r = 0.59, p < 0.01). CONCLUSIONS: A simple and rapid sample pretreatment for the measurement of neopterin in serum has been introduced. The significant positive correlation between neopterin and homocysteine implies that, interference with leukocyte function might be a new possible mechanism for the deleterious effects of homocysteine on vascular function. PMID- 11440727 TI - Development of a quantitative luminometric hybridization assay for the determination of telomerase activity. AB - OBJECTIVES: To develop a quantitative luminometric hybridization assay for the determination of telomerase activity in tissue and cell extracts. DESIGN AND METHODS: Quantification is based on the coamplification of telomeric repeats synthesized by telomerase along with a specifically designed recombinant DNA internal standard (DNA-IS). The DNA-IS has a similar size and the same primer recognition sites as the telomerase DNA products and differs from them only in a central 18 bp sequence. PCR products are captured on microtiter wells via the biotin-streptavidin system and hybridized with two distinct digoxigenin-labeled oligonucleotide probes that are designed to recognize specifically telomerase products and DNA-IS. The hybrids are quantified by a luminometric reaction using an antidigoxigenin antibody conjugated to alkaline phosphatase. The hybridization assay was validated with the MCF-7 breast carcinoma and leukemia K-562 cell lines and a synthetic telomerase product (R(8)). RESULTS: Luminescence ratios for telomerase products and DNA-IS were linearly related to the concentration of the pre-PCR product synthesized by telomerase (R(8)), in the range of 0.0005 to 10 pM. The overall reproducibility of the assay (between-run) varied between 11.3 and 15%. Application of the method in eleven breast tumors showed a great variation in the levels of telomerase enzymatic activity. CONCLUSIONS: The proposed luminometric hybridization assay for the quantitative determination of telomerase enzymatic activity is highly sensitive and can be used for a large scale prospective evaluation of clinical samples. PMID- 11440728 TI - Simultaneous simple and fast quantification of three major immunosuppressants by liquid chromatography--tandem mass-spectrometry. AB - OBJECTIVES: The aim of the current study was to develop a simple, fast and universal method for quantification of any combination of the three major immunosuppressants sirolimus, tacrolimus and cyclosporin in whole blood, using a LC-tandem mass spectrometer (API-2000, SCIEX, Toronto, Canada). METHODS: 250 microL whole blood was spiked with internal standard (ritonavir), and protein precipitated with 350 microL acetonitrile. The sample was centrifuged and 30 microL aliquot was injected onto the HPLC column, where it underwent an online extraction with ammonium acetate. After that the automatic switching valve was activated, changing the mobile phase to methanol and thereby eluting the analytes into the tandem mass spectrometer. The high selectivity of a tandem mass analyzer allows determination of any combination of the three drugs within a 5 min run. RESULTS: Between-day precision was between 2.4% and 9.7% for all analytes at the concentrations tested. Accuracy ranged between 98.8% and 103.2% (n = 20). The method was linear over the measuring ranges of all analytes. Within-run precision was below %CV = 6% for all analytes. Good correlation with other analytical methods was observed. CONCLUSIONS: The simplicity, universality and high throughput of the method make it suitable for application in a clinical laboratory. The method has been implemented in our laboratory for a routine use. PMID- 11440729 TI - Development and clinical application of an immunoassay using intact Helicobacter pylori attached to a solid phase as an antigen. AB - OBJECTIVE: At present, H. pylori homogenates, extracts and recombinant proteins are used as antigens in immunoassays, but significant false positive and negative results are obtained. We attempted to develop an ELISA system using immobilized whole intact H. pylori cells as a solid phase antigen (WIC-ELISA) which specifically recognizes surface antigens. METHODS: Intact H. pylori cells were immobilized on ELISA plates by centrifugation (150 g for 10 min). HRP-labeled antihuman IgG was used as the second antibody. H. pylori-infections were investigated with WIC-ELISA and a conventional method in which H. pylori-extracts were used as antigen in 117 patients. RESULTS: WIC-ELISA showed better characteristics than conventional ELISA in regards to sensitivity (98.9 vs. 90.4%), specificity (95.7 vs. 95.7%), positive predictive value (98.9 vs. 98.8%), negative predictive value (95.7 vs. 71.0%) and accuracy (98.3 vs. 91.5%). CONCLUSIONS: WIC-ELISA is useful for quantification of antibodies against H. pylori surface antigens and as a clinical screening test. PMID- 11440730 TI - Effects of metamizol and magnesium sulfate on enzyme activity of glucose 6 phosphate dehydrogenase from human erythrocyte in vitro and rat erythrocyte in vivo. AB - OBJECTIVE: Effects of metamizol and magnesium sulfate on erythrocyte glucose 6 phosphate dehydrogenase enzyme activity were investigated in in vitro and in vivo conditions. METHODS: For in vitro studies, glucose 6-phosphate dehydrogenase was purified from human erythrocyte and rats were used for in vivo studies. Enzyme activity was determined according to the Beutler method by using a spectrophotometer at 340 nm. RESULTS: The results of in vitro study showed that their mean K(i) values were 6.35 x 10(-3) M for metamizol and 1.32 x 10(-2) M for magnesium sulfate and their inhibition types were uncompetitive. I(50) value was 17 mM for metamizol and 50 mM for magnesium sulfate in in vitro study. In the case of in vivo studies, 200 mg/kg metamizol inhibited the enzyme activity by 40% during the first 1.5 h (p < 0.05), and 225 mg/kg magnesium sulfate significantly inhibited the enzyme activity throughout 24 h (p < 0.01). CONCLUSION: The results of this study suggested that metamizol and magnesium sulfate have significant inhibition effect on the activity of glucose 6-phosphate dehydrogenase enzyme in both in vivo and in vitro. PMID- 11440731 TI - Down regulation of the tumor suppressor gene maspin in breast carcinoma is associated with a higher risk of distant metastasis. AB - BACKGROUND: Maspin (mammary serpin) is a relatively novel serine protease inhibitor with tumor suppressing function in breast cancer. Maspin expression was found in normal breast epithelial cells, but was decreased during tumor progression. Only a few systematic analyses of this phenomenon have been undertaken so far. In this study we developed specific nested reverse transcription polymerase chain reaction (RT-PCR) conditions for the detection of maspin expression in human breast carcinoma and assessed maspin's association with the clinical behavior of primary breast cancers. METHODS: Tumor specimens obtained from 45 primary breast cancer patients were analyzed for maspin expression by a nested RT-PCR assay. Recurrence free survival was evaluated and correlated to maspin expression. RESULTS: The maspin transcript was detected in 29 (64%) breast cancer specimens whereas no expression was found in 16 (36%) cancer specimens. This expression was unrelated to any of the established prognostic factors. However, 6 out of 8 patients who developed distant metastasis (lymph nodes, lung, liver, bone, pleura) within 3 yr after their initial diagnosis showed no maspin expression of the primary breast cancer (p < 0.05). CONCLUSIONS: The lack of maspin expression in breast cancer seems to be associated with a short disease free survival and supports maspin's function as an indicator of tumor aggressiveness and metastatic potential. Nested RT-PCR is a sensitive method to determine maspin expression in human breast cancer tissue. PMID- 11440732 TI - Evaluation of C-reactive protein, a sensitive marker of inflammation, as a risk factor for stable coronary artery disease. AB - OBJECTIVES: Multiple lines of investigations have converged to suggest a prominent role for inflammation in coronary artery disease (CAD). The association of CRP level with active CAD is well documented. The relation, however, between levels of CRP and the presence and extent of stable CAD has seldom been studied in the developing countries. We investigated the association between serum concentration of C-reactive protein (CRP) and angiographically documented coronary artery disease (CAD) in a population of 450 individuals. DESIGN AND METHODS: Ultrasensitive immunoassay was used to measure CRP levels in 284 patients with CAD and 166 control healthy subjects. The association of CRP levels with severity of disease as indicated by > or = 50% stenosis in one vessel (n = 79), two vessels (n = 74), or three vessels (n = 131) was also investigated. RESULTS: CRP levels were greater in the patients with CAD (2.14 (0.88--3.38) vs. 1.45 (0.70--2.55) mg/L, p < 0.0001) than in the respective control subjects. Multiple logistic regression analysis showed CRP as an independent discriminating risk factor for CAD (odds ratio, 3.46, p < 0.001). Significant correlation was identified between CRP levels and severity of CAD (p < 0.0001). Prediction models that incorporated CRP in addition to other established coronary risk factors were significantly better at predicting risk than the models based on the other risk factors alone. CRP level was also an independent predictor of CAD in a subpopulation with normal levels of low density lipoprotein cholesterol (LDL-C < or = 3.4 mmol/L, p < 0.009). CONCLUSIONS: Our findings suggest that CRP has a strong association with stable CAD, as such, the measurement of CRP may improve the coronary risk assessment in Iranian CAD patients. PMID- 11440733 TI - Establishment of reference values of five amniotic fluid enzymes. Analytical performances of the Hitachi 911. Application to complicated pregnancies. AB - OBJECTIVES: to evaluate the analytical performances of the Hitachi 911 analyzer (Roche Diagnostics, F) for gamma-glutamyl-transferase, 5'nucleotidase, amylase, aspartate-amino-transferase and total-alkaline-phosphatase assay in amniotic fluid. To establish reference intervals for these five enzymes throughout gestation. To determine their antenatal diagnostic value. DESIGN AND METHODS: amniotic fluid samples were collected between 14 and 35 weeks of gestation. Weekly numbers ranged from 31 to 92. Techniques developed for serum enzyme assays were applied to amniotic fluid. Two pools of amniotic fluid containing low and high marker levels were used. Within-day and between-day variations were calculated, together with the limits of detection and linearity. Reference ranges were established on 508 amniotic fluid samples, including 23 samples from pregnancies with chromosomal aberrations, 14 with gastrointestinal tract defects and 5 with gastroschisis. RESULTS: the assay technique for total-alkaline phosphatase assay had to be adapted to amniotic fluid, but no adaptation was necessary for the other markers. The within-day CV ranged between 2.2 and 11.2% for low-level samples and from 1.1 to 3.4% for high-level samples. The between day CV ranged from 6.3 to 13.3% for low-level samples and 1.2 to 4.7% for high level samples. Total-alkaline-phosphatase activity fluctuated throughout gestation. Amylase levels and aspartate-amino-transferase levels increased whereas gamma-glutamyl-transferase and 5'nucleotidase levels fell until delivery. All trisomy 18 and trisomy 13 pregnancies, 65% of Down's syndrome pregnancies and all pregnancies with digestive tract defects were associated with marked changes in the level of at least one enzyme. CONCLUSION: The Hitachi 911 is suited to rapid, reliable quantification of amniotic fluid enzymes with only minor adaptation. Useful reference intervals can be obtained throughout gestation. Gamma-glutamyl-transferase, 5'nucleotidase, total-alkaline-phosphatase and amylase assay can help to confirm echographic evidence of bowel disorders and thereby improve patient management especially in case of gastroschisis. PMID- 11440734 TI - New scope in angiogenesis: role of vascular endothelial growth factor (VEGF), NO, lipid peroxidation, and vitamin E in the pathophysiology of pre-eclampsia among Egyptian females. AB - OBJECTIVES: The purpose of this study was to investigate the role of VEGF, NO, MDA, and Vitamin E in the pathophysiology of preeclampsia (PE) among Egyptian women. PATIENTS AND METHODS: Our study included 20 pregnant women with mild PE, 40 pregnant women with severe PE, 20 normal control women and 20 normal control pregnant women. Plasma from all women were subjected to estimation of VEGF using ELISA kit, estimation of nitrates (as end product of NO) and MDA by chemical methods and measurement of Vitamin E using HPLC. RESULTS: The obtained data from this study revealed that, VEGF may be used as a useful marker for PE which is elevated in plasma of women with PE (mild and severe) compared to controls. VEGF could differentiate between PE and controls at a cut-off value of: 70 ng/mL with a sensitivity of 95% and a specificity of 90%. Moreover, at a cut-off value of 250 ng/mL, VEGF could discriminate mild PE from severe PE with a sensitivity of 77.5% and a specificity of 85%. With respect to nitrate levels, they were significantly elevated in normal pregnant women compared with normal controls. Meanwhile, nitrate levels showed a highly significant decrease in PE in relation to both controls, explaining the vasoconstriction and high blood pressure occurring in preeclamptic women. Correlation analysis revealed significant inverse correlation between nitrate levels and VEGF. Lipid peroxidation as indicated by MDA was enhanced in preeclamptic women as compared to healthy controls. The increase in MDA levels was correlated to the severity of PE. A highly significant fall in vitamin E as an antioxidant-scavenging vitamin was seen in preeclamptic women as compared to the controls. Also, a significant drop in the vitamin level was found in severe PE in comparison to mild PE. The oxidant/antioxidant correlation was illustrated by the inverse correlation between vitamin E and MDA(r = -0.837, p = 0.000). CONCLUSION: The findings from this study indicated that VEGF may be used as a clinically useful biomarker for PE for better diagnosis, to differentiate between mild and severe PE, and may also be a target for therapeutic intervention of PE. The increase in levels of VEGF together with reduction in NO levels could confirm the existence of vascular reactivity and endothelial disturbance in PE. The decrease in the vasodilator, NO may in part lead to the vasospasm of preeclampsia, which consequently produces hypoxia. The latter may be a cause of an elevation of VEGF (angiogenic factor) levels in PE. Meanwhile, elevated levels of lipid peroxides together with impaired antioxidant defense mechanisms may play a role in the etiology of PE. PMID- 11440735 TI - Reference interval of D-dimer in pregnant women. PMID- 11440736 TI - Lipid and protein oxidation in erythrocyte membranes of hypercholesterolemic subjects. PMID- 11440737 TI - Oxidative stress of platelets and thrombocytopenia in patients with vivax malaria. AB - Oxidative stress and antioxidative capacity of platelets and the relationship with thrombocytopenia were determined in patients with vivax malaria and compared with those of healthy subjects. Whole blood thrombocyte count, platelet superoxide dismutase and glutathione peroxidase activities of patients with vivax malaria were lower and platelet lipid peroxidation levels were higher in patients than those of healthy subjects. There was an important negative correlation between whole blood thrombocyte count and platelet lipid peroxidation level. The antioxidative mechanisms of thrombocytes were insufficient in malaria patients and caused oxidative stress. The oxidative damage of thrombocytes might be important in the ethiopathogenesis of thrombocytopenia occurring in malaria. PMID- 11440738 TI - Sexual dysfunction in multiple sclerosis: a 2-year follow-up study. AB - BACKGROUND AND OBJECTIVE: Sexual dysfunction severely affects the quality of life of patients, but longitudinal studies of sexual function in multiple sclerosis are lacking. We performed a study on a group of patients with multiple sclerosis to evaluate the change in sexual function and to examine the relationship between sexual dysfunction and other clinical variables over time. METHODS: A 2-year follow-up study on 99 patients with definite multiple sclerosis. Information on sexual and sphincteric disturbances have been collected through face-to-face structured interviews. Disability, independence, cognitive performances and psychological functioning have also been assessed. Spearman rank correlation analysis corrected for multiple comparisons, and linear regression analysis have been performed to test variables relationship and remove the effect of potential confounding covariates. RESULTS: The proportion of patients with sexual dysfunction remained over 70% and did not change during the 2-year follow-up, but the extent and number of symptoms increased significantly The number of symptoms of sexual dysfunction did not change significantly after an exacerbation. Significantly, more patients than before the study resorted to counseling and discussed with doctors of sexual matters. In the univariate analysis, changes in sexual function over time correlated with changes in bladder function (r=0.47, p<0.0001) and EDSS score (r=0.41, p<0.0001), but the multivariate analysis demonstrated that only bladder dysfunction was independently related to sexual dysfunction (R=0.36, p=0.003) when the effect of psychological factors were removed. CONCLUSIONS: Symptoms of sexual dysfunction increase in significance and number over time in patients with multiple sclerosis. Relapses did not influence the number of symptoms of sexual dysfunction, but a worsening of pre-existing symptoms cannot be excluded. The change of sexual function appears to be independently associated to bladder dysfunction. PMID- 11440739 TI - Cyclosporin A treatment modulates cytokine mRNA expression by inflammatory cells extracted from the spinal cord of rats with experimental autoimmune encephalomyelitis induced by inoculation with myelin basic protein. AB - In Lewis rats, treatment with high doses of cyclosporin A (CsA) suppresses clinical signs of experimental autoimmune encephalomyelitis (EAE), although disease occurs when treatment is ceased. Treatment with low doses of CsA causes EAE to take a chronic relapsing course. We have previously shown that CsA treatment causes a decline in the number of T cells and increased inflammatory cell apoptosis in the spinal cord. The present study was undertaken to assess whether CsA therapy also modulates cytokine mRNA expression by inflammatory cells in the spinal cord of rats with EAE, looking for changes that might contribute to the observed effects of CsA on the course of EAE. EAE was induced in Lewis rats by inoculation with myelin basic protein and adjuvants. At the peak of neurological signs, on day 14 after inoculation, rats were given a single intraperitoneal injection of saline, or CsA at a dose of 8, 16, 32 or 64 mg/kg. The next day, rats were sacrificed, the spinal cords removed, inflammatory cells were extracted from the cords, and mRNA isolated from these cells. Expression of cytokine mRNA was assessed by semi-quantitative reverse transcription polymerase chain reaction (PCR) and by quantitative real-time PCR. With both techniques, we found that CsA suppressed the expression of interferon-gamma mRNA and interleukin 2 (IL-2) mRNA. With real-time PCR, we found that CsA caused significantly increased expression of transforming growth factor-beta mRNA. With the different techniques, we observed no consistent pattern of alteration of expression of interleukin-10 or interleukin-4 mRNA. It is possible that these changes in cytokine mRNA expression contribute to the modulation of the clinical course of EAE that is produced by CsA treatment. PMID- 11440740 TI - Differential recording of upper and lower cervical N13 responses and their contribution to scalp recorded responses in median nerve somatosensory evoked potentials. AB - To distinguish the different origins of cervical N13 potentials in median nerve somatosensory evoked potentials (SSEPs), cervical N13 potentials were recorded by two different montages. The abnormal patterns of the SSEPs were compared to the abnormal evoked spinal cord responses (ESCPs) recorded from posterior epidural space in 13 patients with various cervical lesions. SSEPs from the posterior cervical surface were recorded from the mid-cervical level with anterior neck reference (Cv5-AN) and from the upper cervical level with inion reference (Cv2 IN). Scalp responses were recorded from the parietal region contralateral to the stimulating side with non-cephalic reference (shoulder contralateral to stimulating side). ESCPs were recorded from the posterior epidural space using catheter electrodes or needle electrodes inserted into the ligamentum flavum. Lower cervical N13 (LC-N13) recorded from the Cv5-AN montage showed similar latency to upper cervical N13 (UC-N13) recorded from the Cv2-IN montage. The latency of the early part of the P13-P14 complex in the scalp montage was similar to that of the UC-N13 and the negative peak latency of the ESCPs recorded at the C2-3 level. Attenuation of the LC-N13 and relatively preserved UC-N13 and P13-P14 were characteristic in patients with cervical syringomyelia and compression cervical myelopathy at the mid-cervical levels. Attenuation of the UC-N13 with normal LC-N13 was characteristic in patients with cervical spondylotic myelopathy who showed conduction blockade of the ESCPs at the C3-4 level. In a patient with schwannoma at the C1-2 level, conduction blockade of the ESCPs was observed at the C1-2 level. P13 was normal but P14 was prolonged. UC-N13 and P13 latencies were similar to the negative peak latency of the ESCPs at the C2-3 level. We demonstrated that two different cervical N13 potentials can be recorded by two different montages and they represent different behavior in various spinal cord lesions. In addition, at least the early part of the P13-P14 complex originates in the upper cervical region. To distinguish two different cervical N13, it is useful to detect not only the cervical pathology but also the symptomatic cervical cord compression level in patients with cervical myelopathy. PMID- 11440741 TI - Increased interleukin-6 of skin and serum in amyotrophic lateral sclerosis. AB - Studies of skin in amyotrophic lateral sclerosis (ALS) have demonstrated morphological and biochemical alterations. Interleukin-6 (IL-6) has been suggested to have a trophic effect in nerve cells and to have a direct pathogenic role in neurodegenerative central nervous system (CNS) disorders. However, little is known concerning IL-6 in ALS patients. We examined IL-6 immunoreactivity of biopsy specimens of skin and measured serum IL-6 levels from 11 ALS patients and 11 diseased control subjects. IL-6 immunoreactivity was markedly positive in the epidermis and dermal blood vessels and glands and was moderately positive in the reticular dermis in all ALS patients. These optical densities for IL-6 immunoreactivity in ALS patients were significantly higher than in control subjects, and were significantly increased with duration of illness. Serum IL-6 levels were detected in 8 (73%) of 11 ALS patients compared with only 1 (9%) of 11 diseased control subjects. Serum IL-6 levels were significantly correlated with duration of illness in ALS patients and immunoreactivity of IL-6 of the epidermis. These data suggest that the increased levels of serum IL-6 may reflect an increased IL-6 immunoreactivity of skin in ALS patients. PMID- 11440742 TI - Infratentorial hypointense lesion volume on T1-weighted magnetic resonance imaging correlates with disability in patients with chronic cerebellar ataxia due to multiple sclerosis. AB - In multiple sclerosis (MS), hypointense lesions on T1-weighted magnetic resonance imaging are thought to represent areas of tissue disruption and axonal loss. In previous studies of MS patients, infratentorial T1 hypointense lesions were found to be rare. In MS patients selected to have chronic cerebellar ataxia, we have determined the extent of infratentorial T1 hypointense lesions and their relationship with disability. We recruited nine patients with chronic cerebellar ataxia due to MS. An expanded disability status scale (EDSS) assessment was performed on each. The patients' brains were then imaged with axial-oblique dual echo fast spin-echo and contrast-enhanced T1-weighted conventional spin-echo sequences. The number and total volume of infratentorial high-signal lesions on T2-weighted images and infratentorial hypointense lesions on T1-weighted images were calculated by a blinded observer using a computer-assisted contouring technique. A total of 96 infratentorial high-signal lesions were present, of which 62 (64.6%) appeared isointense and 34 (35.4%) hypointense with respect to the surrounding brain substance on the T1-weighted images. There was a median of 3 (range 0-10) and median volume of 0.43 ml (range 0-0.85 ml) infratentorial T1 hypointense lesions per patient. The EDSS score correlated with both the number (r=0.68, p=0.043) and the volume per patient (r=0.89, p=0.001) of infratentorial T1 hypointense but not T2 high-signal lesions. Infratentorial T1 hypointense lesions are often seen in patients with MS and chronic cerebellar ataxia. They may play a significant role in the disability suffered by these patients. PMID- 11440743 TI - Reduced pre-movement facilitation of motor evoked potentials in spinocerebellar degeneration. AB - OBJECTIVES: The aim of this study was to clarify the cerebellar effects on the motor area of the cerebral cortex and abnormal control mechanisms of voluntary movement in spinocerebellar degeneration (SCD). We used transcranial magnetic stimulation (TMS) to study the change in the motor evoked potentials (MEPs) before voluntary movement (pre-movement facilitation) in patients with SCD. SUBJECTS AND METHODS: Pre-movement facilitation of MEPs in subjects' muscles was observed during their thumb movement intention. Patients with SCD, who showed cerebellar signs, without pyramidal or extrapyramidal signs, were examined. TMS was applied randomly during the interval between the "go" signal and the onset of voluntary EMG. The MEPs were recorded from the abductor brevis pollicis muscle. RESULTS: Patients with SCD showed a delay of task performance. In control subjects, the amplitude of MEPs was significantly facilitated (increased) prior to voluntary movement. In patients with SCD, pre-movement facilitation of the amplitude of MEPs was significantly decreased in the study with subthreshold TMS. CONCLUSIONS: Disturbance of pre-movement facilitation in SCD may indicate incomplete cerebellar regulation of voluntary movements. PMID- 11440744 TI - Patients with Multiple Sclerosis and concomitant uveitis/periphlebitis retinae are not distinct from those without intraocular inflammation. AB - Recent models of experimental autoimmune encephalomyelitis (EAE) have indicated that antigens co-expressed in the retina and uvea might be of pathogenetic relevance in Multiple Sclerosis (MS). We investigated the clinical spectrum and magnetic resonance imaging of 11 MS patients with concomitant uveitis, and determined the frequency of clinically silent intraocular inflammation in a prospective series of 50 patients. Two of the 11 patients had panuveitis, seven had anterior, and the remaining two had intermediate uveitis. The onset of uveitis preceded that of neurological symptoms by a mean of 8.5 years (range 1 20). None of the 50 MS patients studied prospectively by using slit lamp examinations and dilated funduscopy showed any evidence of uveitis but six patients had signs of retinal inflammation ("periphlebitis retinae"). Cranial MRI did not reveal "atypical" lesional distribution in MS patients with uveitis or periphlebitis retinae. No correlation between the type of MS and uveitis, or between the degree of neurological disability and the type of uveitis was found. PMID- 11440745 TI - SPECT imaging of the dopamine transporter with [(123)I]-beta-CIT reveals marked decline of nigrostriatal dopaminergic function in Parkinson's disease with urinary dysfunction. AB - We studied a correlation of urinary dysfunction with nigrostriatal dopaminergic deficit in Parkinson's disease (PD) by single-photon emission computed tomography (SPECT) imaging of dopamine transporter with [123I]-2beta-carbomethoxy-3beta-(4 iodophenyl)tropane (beta-CIT). Eleven patients were enrolled in the study, including four men and seven women, with a mean age of 64 years. Seven patients had urinary symptoms 1-5 years after the onset of motor disorder, which included nighttime frequency in six, urinary retardation in four, daytime frequency in one and urge urinary incontinence in one. Using a SPECT camera, the ratio specific to nondisplaceable [123I]-beta-CIT uptake, designated as "striatal V3" was obtained in the caudate, anterior and posterior putamen 24 h after the tracer injection. The striatal V3 was compared in patients with and without urinary dysfunction, and between men and women, using unpaired Student's t-test. Correlation of motor dysfunction and duration of illness with urinary dysfunction, was also analyzed. In the patients, there was a reduction of [123I]-beta-CIT binding in the striatum on both sides, particularly in the putamen contralateral to the affected body side. The striatal V3 of the caudate (p<0.01, Rt; p<0.05, Lt), anterior putamen (p<0.05, Rt) and posterior putamen (p<0.05, Rt) in patients with urinary dysfunction was significantly reduced than those without urinary dysfunction. No sex difference was seen in reduction of [123I]-beta-CIT binding. Urinary dysfunction in PD was more common in patients with higher Unified Parkinson's Disease Rating Scale (UPDRS) score, higher Hoehn-Yahr grade, but not in those with longer duration of disease, although there was no statistical significance. It is likely that our results reflect the association of urinary dysfunction and degeneration of the nigrostriatal dopaminergic cells in PD. PMID- 11440746 TI - Experimental influenza B viral myositis. AB - To investigate the pathogenesis of influenza myositis in animals, juvenile BALB/c mice were inoculated with influenza B/Lee virus intramuscularly into the right quadriceps muscle. Chicken normal allantoic fluid (NAF) or phosphate-buffered saline (PBS) was injected into the left quadriceps of control mice and in some virus-infected mice. Serum creatinine phosphokinase (CPK) levels rose significantly on days 1 and 2 post-inoculation (PI) in only virus-inoculated mice. On days 2 and 3 PI, right quadriceps muscles developed scattered foci of a predominantly mononuclear inflammation in the perimysial connective tissue often adjacent to degenerating or necrotic muscle fibers. Immunofluorescent staining with specific anti-influenza B virus antisera showed muscle fibers that contained specific staining in nuclei and adjacent cytoplasm. Skip areas of staining within muscle fibers suggested that not all muscle nuclei within an individual muscle fiber were infected. A continuous fall in infectious virus titer in the right quadriceps muscles suggested the initial virus inoculum became inactivated and progeny virions were not produced. Left quadriceps muscle never had muscle necrosis or endomysial inflammation, specific staining of viral antigen, virus isolation, or viral RNA detected by the reverse transcriptase polymerase chain reaction assay. These findings support the hypothesis that a non-permissive influenza viral infection can develop in murine skeletal muscle that can damage specific nuclear domains of muscle fibers producing muscle degeneration or necrosis. A similar type of muscle infection may develop in humans that occasionally develop focal myositis during influenza. PMID- 11440747 TI - Deafness associated with vertebrobasilar insufficiency. AB - The incidence of auditory disturbances in vertebrobasilar insufficiency (VBI) is considered much rarer than vestibular disturbances, but no routine audiometric test results for VBI patients have been published. To determine the incidence of deafness associated with VBI and the sites predominantly involved in deafness, we studied 70 vertiginous patients with VBI using a routine audiological examination and magnetic resonance imaging (MRI). MRI detected a lacunar infarct involving the cochlear nuclei in one patient, but lacunae did not involve central auditory structures in the other patients. Twenty-five patients experienced tinnitus, deafness, or both, during vertigo episodes. Audiological examinations confirmed the development of unilateral deafness in 15 (21%) patients, involving the cochlea in 14 and cochlear nuclei in one. These findings indicate that hearing loss occurs in approximately one-fifth of VBI patients, much less frequently than vestibular disturbances, and that deafness associated with VBI mainly involves the cochlea. PMID- 11440748 TI - Afferent pathways of sympathetic skin response in spinal cord: a clinical and electrophysiological study. AB - BACKGROUND: Sympathetic skin response (SSR) recording is an established test of sudomotor autonomic functions. However, knowledge of its pathways in spinal cord is putative. OBJECTIVE: This study involved subjects with isolated spinal cord lesions to evaluate the afferent pathways of SSR. METHODS: Clinical examination was done according to standard neurological classification of spinal cord injury. Electrophysiological evaluation included: (1) conventional nerve conduction studies to exclude peripheral nerve lesions, (2) scalp somato-sensory-evoked potentials (SEP) with posterior tibial nerve (PTN) stimulation and (3) SSR recording from palm by stimulating supra orbital nerve (SON) at forehead, and PTN at ankle. Subjects with absent SSR in palm to SON stimulation were excluded. In such patients, the afferent tracts were considered abnormal when SSR was absent in palm on stimulation of PTN. RESULTS: Among 37 subjects (age-28.1+/-12.8 years), the afferent tracts of SSR were affected in 13. Sparing of afferent SSR tracts correlated with preservation of bladder sensations (P<0.01). There was no correlation between SSR and SEP. CONCLUSIONS: Spinal cord lesions frequently involve afferent tracts of SSR. Spinal afferents of SSR are closely related with tracts of bladder sensations and are different from pathways for SEP. PMID- 11440749 TI - Use of statins in CNS disorders. AB - It is well established that 3-hydroxy-3-methyglutaryl coenzyme A (HMG-CoA) reductase inhibitors ("statins") reduce cholesterol levels and prevent coronary heart disease (CHD). Although a causal relation between elevated cholesterol levels and stroke has not been well defined, a number of large secondary prevention studies and meta-analyses have shown that statin therapy reduces stroke in patients with CHD and hypercholesterolemia. In addition to the vascular effects of statins (stabilization of atherosclerotic plaques, decreased carotid intimal-medial thickness), there are increasing data to suggest that these agents have additional properties that are potentially neuroprotective. These include endothelial protection via actions on the nitric oxide synthase system, as well as antioxidant, anti-inflammatory and anti-platelet effects. These actions of statins might have potential uses in other neurological disorders such as Alzheimer's disease and certain types of brain tumors. PMID- 11440750 TI - Intranasal administration of insulin-like growth factor-I bypasses the blood brain barrier and protects against focal cerebral ischemic damage. AB - BACKGROUND: Insulin-like growth factor-I (IGF-I) has been shown to protect against stroke in rats when administered intracerebroventricularly. However, this invasive method of administration is not practical for the large number of individuals who require treatment for stroke. Intranasal (IN) delivery offers a noninvasive method of bypassing the blood-brain barrier (BBB) to deliver IGF-I and other neurotrophic factors to the brain. Here, we demonstrate for the first time the therapeutic benefit of IN IGF-1 in rats following middle cerebral artery occlusion (MCAO). METHODS: A blinded, vehicle-controlled study of IN IGF-I was performed using the intraluminal suture occlusion model. Rats were randomly divided into vehicle-control, 37.5 and 150 microg IGF-I-treated groups. Treatments occurred at 10 min after onset of 2 h of MCAO, and then 24 and 48 h later. Four neurologic behavioral tests were performed 4, 24, 48 and 72 h after the onset of MCAO. Corrected infarct volumes were evaluated 72 h after the onset of MCAO. RESULTS: Treatment with the 150 microg IGF-I significantly reduced the infarct volume by 63% vs. control (p=0.004), and improved all the neurologic deficit tests of motor, sensory, reflex and vestibulomotor functions (p<0.01). However, the 37.5 microg dose of IGF-I was ineffective. CONCLUSION: While IGF-I does not cross the BBB efficiently, it can be delivered to the brain directly from the nasal cavity following IN administration, bypassing the BBB. IN IGF-I markedly reduced infarct volume and improved neurologic function following focal cerebral ischemia. This noninvasive, simple and cost-effective method is a potential treatment for stroke. PMID- 11440751 TI - Postencephalitic pure anomic aphasia: 2-year follow-up. AB - We report a patient with pure anomic aphasia following encephalitis. Brain magnetic resonance imaging (MRI) revealed bilateral temporal lesions, and subsequent focal atrophy in the left anterior inferior temporal lobe. Over the course of a 2-year follow-up, the patient's naming difficulty persisted without other dysfunction of language or memory. These observations indicate a contribution of the left anterior inferior temporal region to object naming. PMID- 11440752 TI - Choreiform movements in spinocerebellar ataxia type 1. AB - We describe the unusual case of a 51-year-old woman with spinocerebellar ataxia type 1 (SCA1) who showed choreiform movements in addition to cerebellar ataxia. To date, extrapyramidal signs including involuntary movements have been rarely reported in SCA1. Surface electromyogram in our patient revealed grouped discharges whose duration was longer than that of chorea observed in HD, indicating that the involuntary movements represented choreoathetosis rather than pure chorea. These choreiform movements have not been seen in non-hereditary spinocerebellar ataxia. Therefore, if "sporadic" cases of cerebellar ataxia show such movements, the possibility of genetic origin of the ataxia is high and a surveillance of various forms of hereditary spinocerebellar ataxia including SCA1 is required. PMID- 11440754 TI - Laterally directed arm movements and right unilateral neglect after left hemisphere damage. AB - Signs of unilateral neglect for events occurring in one hemispace most often result from right hemisphere lesions. Right unilateral neglect after left hemisphere damage is much rarer, and has received less attention. The present study explores the relationships between right unilateral neglect and asymmetries in producing laterally directed arm movements in the horizontal plane in left brain-damaged (LBD) patients. Participants produced right- or left-directed arm movements with their left arm in response to centrally located visual stimuli. Results showed that LBD patients with signs of right unilateral neglect were consistently slowed when producing arm movements toward the right (neglected) side, as compared to left-directed movements. Taking into account patients with and without signs of neglect, this directional asymmetry positively correlated with a reaction-time measure of perceptual spatial bias. These findings stand in contrast with previous results obtained with the same experimental paradigm in right brain-damaged patients, in whom a consistent slowing of leftward-directed movements was rare and apparently unrelated to the presence and severity of left neglect. These conflicting results are discussed with respect to the hypothesis that different mechanisms may underlie left and right unilateral neglect. PMID- 11440755 TI - A particular difficulty in discriminating between mirror images. AB - We investigated the selective impairment of mirror image discrimination in a patient with bilateral parieto-occipital lesions (FIM). We report a difficulty with the discrimination between mirror images more selective than has been previously reported (Turnbull OH, McCarthy RA. Failure to discriminate between mirror-image objects: a case of viewpoint-independent object recognition? Neurocase 1996;2:63). FIM was asked to judge, in five same/different experiments, whether pairs of simultaneously presented line drawings of objects were identical. FIM demonstrated only a minor impairment in discriminating between orientations in the picture plane but was at chance in making discrimination between mirror images. An experiment with normal observers established that our results were not due to differences in task difficulty. Two further experiments investigated the effects of rotation on the discrimination of letters and geometric shapes. FIM's impairment extended to geometric shapes but not to letters. These results would be consistent with the preservation of an abstract representation for object recognition that did not code the difference between mirror image views. PMID- 11440756 TI - Right medial thalamic lesion causes isolated retrograde amnesia. AB - Pervasive retrograde amnesia without anterograde memory impairment has rarely been described as a consequence of circumscribed brain damage. We report this phenomenon in a 33 yr-old, right-handed man (JG) in association with the extension in the right thalamus of a previously small, bilateral thalamic lesion. JG presented with a dense amnesia for autobiographical material more than a few years old, with some sparing of recent memories. Furthermore, he was completely unable to recognise famous people or world events. Many other aspects of semantic knowledge were intact and there was no evidence of general intellectual impairment, executive dysfunction or loss of visual imagery. Magnetic resonance imaging revealed an acute lesion in the right thalamus and two small, symmetrical, bilateral non-acute thalamic lesions. Follow-up neuropsychological assessment indicated a stable pattern of impaired retrograde and spared anterograde memory over 18 months and psychiatric assessments yielded no evidence of confabulation, malingering or other symptoms to suggest psychogenic amnesia. JG's profile indicates that the division of declarative memory into just two categories - episodic and semantic - is inadequate. Rather, his case adds to the growing body evidence to suggest that world knowledge pertaining to people and events is stored or accessed similarly to autobiographical information and differently from other types of more general factual knowledge. We hypothesize that the right mediodorsal thalamic nucleus and immediately surrounding regions comprise the central processing mechanism referred to by McClelland (Revue Neurologique, 150 (1994) 570) and Markowitsch (Brain Research Review, 21 (1995) 117) as responsible for inducing and co-ordinating the recall of these sorts of cortically stored memory engrams. PMID- 11440757 TI - Neural substrates for recognition of familiar voices: a PET study. AB - Identification of familiar people is essential in our social life. We can identify familiar people by hearing their voices as well as by viewing their faces. By measuring regional cerebral blood flow (rCBF) by positron emission tomography (PET), we identified neural substrates for the recognition of familiar voices. The brain activity during discrimination of voices of the subjects' associates and friends from those of unfamiliar people was compared with that during an analogous discrimination of their own voice from unfamiliar voices as well as during vowel discrimination. The left frontal pole, right temporal pole, right entorhinal cortex, and left precuneus were activated to a greater extent during discrimination of familiar voice than during control discriminations, suggesting that these brain regions are involved in the recognition of familiar voices. Furthermore, the adjusted values of rCBF in the left frontal pole and right temporal pole correlated with the number of subjects' correct identification of familiar voices. The present results suggest that these two regions are coactively associated with matching the currently heard voice to familiar voices in one's memory. PMID- 11440758 TI - Interaction of hand use and spatial selective attention in children. AB - It has been argued that concurrent motor action can modulate visual spatial attention. The visual spatial biases of adult patients with unilateral neglect, for example, can be ameliorated by simultaneous use of the contralesional hand. Such improvements are most dramatic when the contralesional hand is moved within contralesional space. To date, evidence of such an interaction in neurologically healthy individuals has not been presented. Line bisection is a simple task that is sensitive to attentional spatial bias. When young children are asked to bisect horizontal lines using their right hands, they show a reliable, if small, bias that is consistent with the pattern seen in adult neglect. This bias is reversed when the left hand is used. Here, we show that these effects are significantly modulated by the location of the movements relative to the body mid-line - specifically that the conjunction of hand movements within ipsilateral space is necessary for the previously reported pattern to be observed. We further demonstrate that these effects are not present in the bisections of neurologically healthy adults. In a final study, we examined whether the hand movement effects seen in children's line bisections would persist in a purely visual task (that is when the movements were made irrelevant to the response). Again, significant modulation of children's perception by concurrent hand movements - and the relative location of those movements - was observed. The theoretical and clinical implications of the results are discussed. PMID- 11440759 TI - Performance consistency of normal observers in forced-choice tachistoscopic visual line bisection. AB - Pseudoneglect (PN) refers to the leftward error exhibited by normal observers on line bisection tasks (Bowers and Heilman, Neuropsychologia, 18, (1980) 491-8). Although a thorough review of the literature has shown PN to be relatively robust (Jewell and McCourt, Neuropsychologia, 38, (2000) 93-110), controversy remains concerning the reliability of the phenomenon, with some studies reporting a relatively high incidence of normal subjects with rightward bisection errors. The present experiment assesses the consistency of bisection performance in normal young observers. Right-handed subjects (N=22) participated in a tachistoscopic forced-choice line bisection task. Each subject participated in 7-16 experimental sessions separated by at least 24 h (total bisection measurements=317). Individual bisection performance could thus be evaluated with respect to within subject variability measures. An eyetracker recorded gaze position during the task in one session. A highly significant mean group bisection error of -0.26 degrees (P<0.001) was obtained (left negative), and individual subject means ranged from -0.55 degrees to +0.03 degrees. Of the 317 total bisection measurements, 9% (28) deviated rightward. Significant (P<0.05) mean leftward errors occurred in 91% (20/22) of subjects. Mean bisection error in two subjects was not significantly different from zero. No subject possessed a significant rightward error. Mean gaze deviation from screen (and line) center ranged from +/ 0.9 degrees, and was positively correlated (P<0.05) with bisection error. It is concluded that forced-choice tachistoscopic line bisection measures are highly reliable; a mean correlation of +0.87 exists between mean error based on 15 trials and means estimated from a random sample of only two trials. The incidence of true rightward bisection error in the population of normal right-handed subjects is thus estimated to be less than 5%. PMID- 11440760 TI - Consistency among different tasks of left-right asymmetries in lines of fish originally selected for opposite direction of lateralization in a detour task. AB - Lines of fish, Girardinus falcatus preferentially turning rightward (RD) or leftward (LD) when facing a dummy predator visible behind a barrier have been obtained through selective breeding. To check whether lateralization was maintained in other behavioral responses, five different tests were carried out. They comprised measures of (1) turning direction in a T-maze; (2) proportion of clockwise and anticlockwise direction of rotation in a circular arena; (3) preferential eye use by females during shoaling behavior (i.e. while looking at their own mirror image reflection); (4) preferential eye use by males during sexual behavior (i.e. while turning around a barrier to join a group of females); and (5) preferential eye use by males during agonistic behavior (i.e. while attacking a rival visible in a mirror). In all five tests the two selected lines showed opposite direction of lateralization. Results thus indicate that behavioral asymmetries in the detour test are predictive of lateralization in other types of behavioral tests. Moreover, results show that RD and LD fish have a similar but left-right reversed pattern of subdivision of cognitive/ behavioral functions, which is suggestive of a similarly left-right reversed (mirror image) brain organization. PMID- 11440761 TI - Functional differences in the semantic processing of concrete and abstract words. AB - There is considerable debate as to whether the semantic system is a unitary one in which meanings are available in a peculiar, perceptual-free format, or whether it is functionally segregated into anatomically discrete, modality-specific but semantic regions. In the former case, concrete and abstract words should not differ in the amount of activation of semantic areas. Neuroimaging studies in this field are, however, far from conclusive, and one reason for this may be that the degree of imageability of the stimuli - probably a crucial variable - has not been considered. Recognition Potential (RP) reflects semantic processing and appears to originate in basal extrastriate regions involved in semantic processing. In this study, we compared the RP of concrete and abstract words that actually differ in their degree of imageability. Results indicate that the semantic processing areas in which the RP originates display a higher activation for concrete (more imageable) material, but that abstract material also evokes a notably larger RP component compared with pseudowords or unpronounceable letter strings. Accordingly, the study appears to suggest that there is no full functional segregation of the semantic systems. Rather, our data support the existence of a semantic system that is specialised in concrete, imageable material, and that is also activated, though to a lower extent, by abstract material. PMID- 11440762 TI - Disruption of estimation of body-scaled aperture width in Hemiparkinson's disease. AB - A group of patients with left-sided symptoms of Parkinson's disease (LPD) was compared with a group of patients with right-sided symptoms (RPD) and with a group of healthy age-matched controls on body-scaled judgements of aperture width. Participants judged whether or not they would fit through a life-sized schematic doorway shown on a large screen. A staircase technique was used to find the door width for which 50% of the judgements were positive. The ratio between this measure and the width of the participant's body at the shoulders (the aperture to shoulder - A/S ratio) was calculated. The A/S ratio was approximately 1.5 in the LPD group, approximately 0.9 in the RPD group, and approximately 1.1 in the control group, suggesting that the visual representation of the doorway (or that of its relationship to perceived body-size) is compressed in LPD (and perhaps expanded in RPD). The A/S ratios were invariant with viewing distance (0.6 or 1.5 m), the contrast polarity of the display (white on dark, or vice versa) and the doorway surround (blank, or vertical or horizontal stripes). The findings are discussed with reference to the neural representation of external space and of the body, and to the motor problems of Parkinson's disease. PMID- 11440763 TI - Cerebral blood flow velocity changes during dichotic listening with directed or divided attention: a transcranial Doppler ultrasonography study. AB - Simultaneous bilateral transcranial Doppler ultrasonography (TCD) monitoring of blood flow velocity (BFV) in the middle cerebral arteries was performed in 28 normal right-handed volunteers during linguistic dichotic listening tasks to investigate the effect of hemispheric specialisation and allocation of attention. A control task that required the repetition of monaurally presented words was followed by three randomised dichotic listening tasks in which the subjects were instructed to direct their attention to the word stimuli of the right ear, the left ear, or to divide their attention between both ears. Behavioural data indicated that the subjects used the required attentional strategies. A significant right ear advantage was not obtained in the divided attention condition due to a ceiling effect. Each task resulted in a significant bilateral increase in BFV. We found no significant lateralisation of BFV change for any of the tasks. We noted marked differences in BFV change between the different conditions that were significant in the right hemisphere, showed a borderline significance in the left hemisphere and appeared to be related with the difficulty of the task. We conclude that the hemodynamic changes caused by attentional strategies or hemispheric specialisation in processing dichotic stimuli over and above the effect of bilateral auditory stimulation, are too subtle to be detected as lateralised changes in BFV. Functional TCD could be used for the evaluation of a task's workload relative to other tasks, and may contribute to elucidate the role of the right hemisphere in attention and arousal. PMID- 11440764 TI - Controlling attentional priority by preventing changes in oculomotor programs: a job for the premotor cortex? AB - Abruptly presented items capture attention automatically so they constitute the first items to be examined [Yantis and Jonides, Journal of Experimental Psychology: Human Perception and Performance, 1984;10:601; Jonids and Yantis, Perception and Psychophysics, 1988;43:346; Theeuwes, Perception and Psychophysics, 1992;51:599; Theeuwes, Journal of Experimental Psychology: Human Perception and Performance, 1994;20:799]. This attentional priority can be controlled in a top-down manner by directing attention towards the locus of interest [Yantis and Johnson, Journal of Experimental Psychology: Human Perception and Performance, 1990;16:812; Theeuwes. Perception and Psychophysics, 1991;49:83; Miller, Perception and Psychophysics, 1989;45:567; Folk et al., Journal of Experimental Psychology: Human Perception and Performance, 1992; 18:1030]. The premotor theory of attention [Rizzolatti et al., Neuropsychologia 1987;25:31; Rizzolatti et al., Attention and Performance XV, 1994, p. 231] assumes that the mechanism responsible for the attentional shifts is strictly linked to that responsible for eye movements, and several studies [Corbetta et al., Society of Neuroscience Abstracts 1997;23:122.12; Nobre et al., Brain 1997;120:515; Theeuwes et al., Journal of Experimental Psychology: Human Perception and Performance, 1999;25:1595] suggested that the premotor cortex plays a role in the control of attention. However, the nature of this involvement is still unclear. We have been asking a patient (RJ) with a damage of the right premotor cortex to decide whether a target had a discontinuity on its right or left side. The absolute location of the target was pre-cued. In Section 2, an interference was observed when a sudden onset occurred in the visual space, suggesting that RJ was not able to control attentional capture. The possibility to attribute this interference to an insufficient focalization of attention or a grouping effect were discarded by Sections 3 and 4, respectively. Section 5 revealed that this interference followed exclusively the onset occurring in the hemifield opposite the one containing the target (meridian effect [Rizzolatti et al., Neuropsychologia 1987;25:31]). The results suggest that the control of attentional capture may be achieved by keeping constant the parameters of the appropriate oculomotor program. PMID- 11440765 TI - An unusual, structurally preserved ovule from the Permian of Antarctica. AB - Anatomically preserved ovules are described from silicified peat of Late Permian age collected from Skaar Ridge in the central Transantarctic Mountains, Antarctica. The small ovules are significant in possessing fleshy apical appendages and a funnel-shaped micropylar extension formed by the sarcotestal layer of the integument, by which they differ from all other Permian ovules described to date. The apical modifications may have functioned in pollination and/or seed dispersal. Similarity with the apical organization of earlier Paleozoic ovules is shown to be superficial, since the analogous structures are developmentally derived from different tissues. Although the ovules occur in rocks in which glossopterids are the only gymnosperms represented, there is insufficient evidence to assign them to a taxonomic group. These ovules are of particular importance because there are so few anatomically preserved gymnosperm reproductive structures known from the Permian and thus provide new data on the diversity of late Paleozoic gymnosperms. PMID- 11440766 TI - Cretaceous (Late Albian) coniferales of Alexander Island, Antarctica. 2. Leaves, reproductive structures and roots. AB - Coniferous foliage from the Albian of Alexander Island, Antarctica, is assigned to the Araucariaceae, Podocarpaceae, and Taxodiaceae based on attached or associated fertile remains. Araucarian foliage represented by Araucaria alexandrensis sp. nov. and A. chambersii sp. nov. is associated with ovulate cone scales described as Araucarites wollemiaformis sp. nov. and A. citadelbastionensis sp. nov., respectively. The Podocarpaceae is represented by Bellingshausium willeyii sp. nov. and the Taxodiaceae by Athrotaxites ungeri, both with attached cones. Sterile foliage is widespread belonging to the form genera Podozamites, Elatocladus, Brachyphyllum and Pagiophyllum. The conifers in this Albian southern high-latitude flora make up ca. 15% of the species diversity. Evidence from leaf litter distribution on palaeosols and leaf morphology suggest that the majority of conifers were large canopy-forming trees, although a few were probably small understorey shrubs. PMID- 11440767 TI - High altitude C(4) grasslands in the northern Andes: relicts from glacial conditions? AB - The altitudinal vegetation distribution in the northern Andes during glacial time differed from the present-day conditions as a result of temperature and precipitation change. New evidence indicate that as a response to a reduced atmospheric partial CO(2) pressure (pCO(2)), the competitive balance between C(3) and C(4) plants have changed. Effects may have remained virtually undetected in pollen records, but can be observed using a stable carbon isotope analysis. Vegetation dominated by C(4) taxa, belonging to the families Cyperaceae (e.g. Bulbostylis and Cyperus) and Poaceae (e.g. Muhlenbergia, Paspalum and Sporobolus), may have been able to replace for a significant part the modern type C(3) taxa (e.g. species belonging to Carex, Rhynchospora, Aciachne, Agrostis, Calamagrostis, and Chusquea). Impact of reduced glacial atmospheric pCO(2) levels and lower glacial temperatures on the composition and the elevational distribution of the vegetation types is discussed. The present high Andean vegetation communities may differ from the glacial equivalents (non-modern analogue situation). We identified dry Sporobolus lasiophyllus tussock grassland and Arcytophyllum nitidum dwarfshrub paramo as the possible relict communities from glacial time. The effect on previous estimates of paleo-temperatures is estimated to be small. PMID- 11440768 TI - Late Holocene rainforest disturbance in French Guiana. AB - Palm swamp forest sediments collected in French Guiana provide new data about late Holocene rainforest. Two cores were collected in 'Les Nouragues' ecological station (4 degrees 05'N, 52 degrees 40'W). The lithology shows two different types of sediment, organic peat in the upper part and oxidized clay with low organic content and lacking pollen in the lower part, both separated by a gravel horizon. Radiocarbon dates show that this gravel horizon could have been deposited between 4500 and 3000yrBP. Pollen analysis carried out on the organic sediments record rainforest disturbances between ca 1520-1380 and 1060-860cal yrBP suggested by the presence of the pioneer tree species Cecropia together with other shade intolerant genera. Cecropia is recorded for a period that lasts between 660 and 320 years. This abnormal duration for presence of a pioneer species in rainforest is explained by brief and repeated changes in the composition of the canopy asssociated to perturbations of the palm swamp. PMID- 11440769 TI - Lateglacial and Early Holocene vegetation history of the northern Wetterau and the Amoneburger Basin (Hessen), central-west Germany. AB - The Lateglacial and Early Holocene vegetation history of the northern Wetterau and Amoneburger Basin, two intra-montane basins in Hessen, central-west Germany, is reconstructed by means of pollen and macrofossil analyses. Regional pollen assemblage zones are defined for the Lateglacial and Early Holocene. After calibration of the radiocarbon dates and establishment of age/depth relationships, the ages of the pollen zone boundaries are calculated. The regional vegetation changes correlate closely with the major fluctuations in the delta18O curve of the Greenland ice cores spanning the same time period. During the early Lateglacial, the open herbaceous vegetation with dwarf shrubs in the northern Wetterau was replaced by woodlands. Initially these woodlands consisted of birch, but after the immigration of pine, mixed forests of pine and birch developed. Soon after its immigration pine became dominant and pine woodlands largely replaced the birch forests. Only on the locally wetter substrates of the river floodplain did Betula stands persist. Gradually the importance of herbaceous communities declined and the pine woodlands lost their open character. During the Lateglacial two regressive phases in the vegetation succession are reflected in the data which are equated with the Older- and Younger Dryas biozones. At the beginning of the Younger Dryas, the forest-limit was lowered and the importance of herbaceous communities increased. Later, pine woodlands thinned and Ericales became part of the vegetation, indicating the development of more acid, nutrient-poor soils. A subdivision of the Younger Dryas biozone into a wetter, colder first part and a drier, warmer second part is suggested. At the beginning of the Early Holocene, pine woodlands became more closed and soils more stabilised. The transition between the Younger Dryas and Preboreal biozones is indicated by a lithological change to organic (-rich) deposits. Betula stands persisted on the locally wetter substrates of the fluvial plains. At the end of the early Preboreal, there was an extension of Betula stands on the river floodplains. This is interpreted as a phase in which climate was temporarily wetter, perhaps a reflection of a short-term climatic oscillation, the so-called Preboreal oscillation. During the Preboreal biozone, ferns became more important in the local vegetation of the mires, and later during the Boreal they formed the undergrowth of swamp forests. During the Boreal biozone, a major expansion of hazel took place in both areas. On the hills and slopes of the northern Wetterau and the Amoneburger Basin, open pine forests and hazel scrub developed with grasses and Pteridium in the field layer. In the river valleys, pine forests were replaced by deciduous mixed forests with oak, elm and later also with lime. PMID- 11440770 TI - Oleaceous laurophyllous leaf fossils and pollen from the European Tertiary. AB - By using cuticular analysis for the first time laurophyllous leaf remains from the European Tertiary have been reliably identified as belonging to the Oleaceae. Despite ecologically determined variation in cuticular structures, Late Miocene leaf material from northern Italy is assigned to a single species, Oleinites liguricus Sachse n. sp. The associated oleaceous pollen grains support that this species has been a prominent component, at least locally, of a laurophyllous forest community. From extensive comparisons with leaf cuticles and pollen of extant Oleaceae it may be concluded, that the fossil leaves and probably corresponding pollen represent the tribe Oleeae and may originate from a plant closely comparable to extant Chionanthus and Fraxinus. A reinvestigation of some Oligocene laurophyllous leaf remains has uncovered two more oleaceous species in the European Tertiary: Oleinites maii (Buzek et al.) Sachse comb. nov., and Oleinites hallbaueri (Mai) Sachse comb. nov. Considering the high diversity of laurophyllous Oleaceae in modern evergreen and mixed mesophytic forests and the common presence of oleaceous pollen in the fossil record it may be expected that cuticular analysis of laurophyllous leaf fossils will lead to the recognition of an increased number of oleaceous taxa in Tertiary leaf assemblages. PMID- 11440771 TI - Biological markers and diagnostic accuracy in the genetics of posttraumatic stress disorder. AB - Family and twin studies suggest a substantial genetic contribution to the etiology of posttraumatic stress disorder (PTSD). Identification of the nature of this genetic contribution should enhance understanding of the pathophysiology of PTSD and suggest improved therapeutic strategies for its treatment. However, a broadly defined phenotype, specific requirement for an environmental exposure and high frequency of comorbid psychiatric illness all complicate genetic studies of PTSD. It is likely that genetic heterogeneity, incomplete penetrance, pleiotropy and the involvement of more than one gene all constitute formidable obstacles to the genetic analysis of PTSD. One way to circumvent these problems is to perform genetic analysis of traits associated with PTSD, rather than PTSD itself, an approach that has been fruitful for other diseases with complex modes of inheritance. Hypothalamic-pituitary-adrenal axis hypofunction, physiologic markers of increased arousal, and increased acoustic startle response are all potential PTSD-associated traits that might be susceptible to genetic analysis. However, the capacity of these traits to distinguish PTSD from non-PTSD patients and their familial pattern must be better defined before they can be employed in genetic studies. PMID- 11440772 TI - Psychiatric symptoms and CAG repeats in neurologically asymptomatic Huntington's disease gene carriers. AB - The putative relationship between the psychiatric profile of a sample of neurologically asymptomatic Huntington's disease gene carriers and CAG repeats was investigated. The psychiatric assessments (by consultant psychiatrist and computerised battery) were undertaken before the genetic testing was carried out. In this way, the informational distortions caused by neurological and cognitive deficits were avoided. The hypothesis that there is a relationship between psychiatric and CAG repeats was tested by seeking direct correlations between psychiatric systems and CAG repeats, and also by correcting the correlation by the number of years above or below the estimated age of onset in Huntington's disease. Scores for irritability and cognitive failures were high in the sample. There was no correlation between any psychiatric variable and CAG repeats. Possible explanations for this lack of correlations are discussed. PMID- 11440773 TI - Risperidone pre-treatment reduces the euphoric effects of experimentally administered cocaine. AB - Pre-clinical research implicates dopaminergic neurotransmission as critical in producing the effects of stimulants. Previous stimulant challenge studies using volunteers treated with dopaminergic antagonists have generally failed to demonstrate reduction of subjective effects. We performed this study to determine whether repeated dosing with risperidone reduced the subjective effects of experimentally administered cocaine. Nine non-treatment seeking hospitalized cocaine-dependent volunteers received 40 mg cocaine IV before and following 5 days of treatment with risperidone, 2 mg per day. Risperidone pre-treatment reduced the self-rated 'high' produced by cocaine. Repeated, rather than single, dosing with a D2 antagonist may be necessary to reduce the subjective effects produced by cocaine. The degree of D2 receptor blockade produced by risperidone appears to be greater than the reduction in euphoric effects produced by cocaine, suggesting that mechanisms other than those involving D2 receptors may be important in drug-induced euphoria. PMID- 11440774 TI - Impaired emotional facial expression recognition in alcoholism compared with obsessive-compulsive disorder and normal controls. AB - Emotional facial expression (EFE) decoding skills have been shown to be impaired in recovering alcoholics (RA). The aim of the present study is to replicate these results and to explore whether these abnormalities are specific to alcoholism using two control groups: non-patient controls (NC) and patients with obsessive compulsive disorder (OC). Twenty-two alcoholic patients at the end of their detoxification process (RA) were compared to 22 OC and 22 NC matched for age, sex and education level. They were presented with 12 photographs of facial expressions portraying different emotions: happiness; anger; and fear. Each emotion was displayed with mild (30%) and moderate (70%) intensity levels. Each EFE was judged on 8 scales labeled happiness, sadness, fear, anger, disgust, surprise, shame and contempt. For each scale, subjects rated the estimated intensity level. RA were less accurate in EFE decoding than OC and NC, particularly for anger and happiness expressions. RA overestimated the emotional intensity for mild intensity level expressions compared with both OC and NC while no significant differences emerged for moderate intensity level expressions. Deficits in EFE decoding skills seem to be specific to RA when compared with OC. Comparison with other psychopathological groups is still needed. Possible consequences of EFE decoding deficits in RA include distorted interpersonal relationships. PMID- 11440775 TI - Lower prolactin bioactivity in unmedicated schizophrenic patients. AB - In previous work, prolactin (PRL) abnormalities of a lower bioassay (BA) to radioimmunoassay (RIA) ratio were found in schizophrenic patients. This line of research was extended in seven male patients with schizophrenia who were neuroleptic-free; seven male control subjects were also studied. PRL values were assessed by RIA and Nb(2) BA techniques. The schizophrenic group had a significantly lower PRL BA as compared to normal controls and a lower PRL ratio of BA/RIA. The lower ratio is consistent with an earlier finding and suggests that schizophrenic patients have different molecular forms of PRL than control subjects. This difference could be due to a disordered tuberoinfundibular dopamine system or the long-term effects of neuroleptic medications. PMID- 11440776 TI - Increased platelet aggregability in major depression? AB - There is compelling evidence that depression constitutes an independent risk factor for cardiovascular morbidity and mortality. As exaggerated platelet reactivity is associated with an increased risk of intra-arterial thrombus formation, we studied platelet aggregability in patients with major depression both before and after 5 weeks of anti-depressant therapy as well as in healthy control subjects. Twenty-two depressed patients and 24 healthy control subjects participated in the study. Washed and rediluted platelets were stimulated with the agonists collagen and thrombin in three concentration steps. Depression was associated with a higher aggregability after stimulation with thrombin in the intermediate concentration and with collagen at the low concentration, with ceiling effects for the other concentrations. After 5 weeks of anti-depressant therapy, aggregability was somewhat less exaggerated, although this effect did not reach statistical significance. We thus conclude that major depression is associated with increased platelet aggregability, which seems to persist even under a marked improvement in depressive symptomatology. This effect may contribute to the increased cardiovascular morbidity in depressed patients. PMID- 11440777 TI - The validity of a short version of the Body Shape Questionnaire. AB - This study investigated the 'construct validity' (which includes the extent to which a test conforms to predictions from previous observations) of a 14-item version of the Body Shape Questionnaire (BSQ) in 75 patients with psychogenic low weight and a history of full or partial anorexia nervosa (AN). Various predictions were confirmed, i.e. ratings on the BSQ (designed to measure concern about body image) were significantly correlated with ratings for the Eating Attitudes Test, the Bulimia Investigatory Test and the Beck Depression Inventory, and with the body mass index at referral. Also, the binge eating/purging subtype of AN was associated with increased body-image psychopathology. In contrast to a previous report using another questionnaire, this association was independent of body weight. The findings indicate that this shorter version of the BSQ can be used to assess body image psychopathology in patients with psychogenic low weight and to confirm an increased concern about body-image in the binge-eating/purging subtype of AN. PMID- 11440778 TI - Lectins in fruits having gastrointestinal activity: their participation in the hemagglutinating property of Escherichia coli O157:H7. AB - BACKGROUND: In fruits with therapeutic properties for antidiarrheal and laxative uses, the presence of lectins may be the bioactive properties that interfere with bacterial adhesion, thought to be competition for glycoside signal sites in the attachment. METHODS: This study identifies lectins in crude extracts from fruits such as Tamarindus indica (tamarind), Spontia vulgaris (plum), Psidium guava (guava), Mangifera indica (mango), Cydonia vulgaris (quince), and Crataegus mexicanus (tejocote). To verify the procedures, extracts from Ricinus communis (castor bean), Glycine max (soybean), Phaseolus vulgaris (beans), Vicia fava (fava bean), and Solanum tuberosum (potato) were used as controls for lectin activity. Both sources of lectins were analyzed to determine their participation in the host-parasite interaction, using as a model the hemagglutinating properties of Escherichia coli O157:H7 (EHA). RESULTS: All extracts showed hemagglutination to group O erythrocytes test (HA) with the exception of mango. Two new galactose-specific lectins were identified from tamarind and guava. When analyzed for participation in EHA, only guava lectins inhibited this, while soybean lectin induced hemolysis; as both lectins bind to galactose, it is probable that their recognition occurs in different domains. Sugars involved in the attachment between Escherichia coli O157:H7 and red cells were identified and again, galactose in addition to mannose was found to be related in EHA. On the other hand, guava lectins also agglutinated E. coli O157:H7, perhaps due to the same galactose-specific lectin or to another type of lectin. CONCLUSIONS: In summary, guava has a galactose-specific lectin that prevents adhesion of E. coli O157:H7 to red cells; this lectin is mediated by galactose. Prevention could also be due to their capacity of agglutinating E. coli by guava lectins. Soybean lectin induced hemolysis only when bacteria was present, but not with floating secretions. This finding showed that guava is a source of lectin that can be explored to prevent adhesion of E. coli to epithelial intestinal cells; contrariwise, soya must be studied to see its participation in the uremia caused during E. coli O157:H7 pathogenesis. PMID- 11440779 TI - The influence of chloroethylclonidine-induced contraction in isolated arteries of Wistar Kyoto rats: alpha1D- and alpha1A-adrenoceptors, protein kinase C, and calcium influx. AB - BACKGROUND: It has recently been reported that chloroethylclonidine (CEC) elicited contraction in tail arteries (alpha(1A)-adrenoceptors) and aorta (alpha(1D)-adrenoceptors) from normotensive and spontaneously hypertensive rats (SHR). This study investigated the relationship between CEC-induced contraction and the role of protein kinase C (PKC) and extracellular Ca(++) influx in tail arteries and aorta from Wistar Kyoto rats (WKY). METHODS: Time-course of CEC induced contraction in endothelium-denuded arteries from Wistar, WKY, and SHR rats was evaluated. In WKY arteries, calphostin C (1 x 10(-6) M) and nitrendipine (1 x 10(-6) M) were used to determine the role of PKC and extracellular Ca(+1) in the contractile response to CEC, respectively. RESULTS: Chloroethylclonidine (1 x 10(-4) M) elicited contraction in tail arteries and aorta from normotensive and hypertensive rats. Maximal response to CEC was similar in tail arteries among strains (approximately 30% of norepinephrine effect), while in aorta CEC elicited a higher contraction in WKY and SHR than in Wistar (59, 86, and 18% of norepinephrine effect, respectively). CEC-elicited maximal contractile responses were reached in 5 min in tail arteries and in 30-45 min in aorta irrespective of the rat strain, suggesting that different intracellular signaling pathways are involved in the contractile response to CEC in these arteries. In WKY tail arteries, calphostin C and nitrendipine blocked CEC-induced contraction while in aorta nitrendipine, but not calphostin C, inhibited CEC action. CONCLUSIONS: This study confirms marked strain-dependent differences in rat aorta responsiveness to CEC and suggests a central role for PKC in response to CEC in tail arteries and for extracellular Ca(+1) influx in aorta. PMID- 11440780 TI - Expression of TCF, TPF/YY1, and the Sp family transcription factors in rabbit endometrium throughout pregnancy. AB - BACKGROUND: TCF, TPF/YY1, and the Sp family are specific transcription factors that bind sequences found within the uteroglobin (UG) gene promoter region that are necessary for transcription. To date, UG gene expression and regulation in vivo are not fully understood. The purpose of this study was to assess the expression patterns of these factors in the rabbit endometrium throughout pregnancy. METHODS: Endometrial nuclear extracts were obtained from female rabbits on days 0, 3, 5, 7, 15, and 28 after mating. Transcription factor expression was assessed by DNA-protein binding assays using endometrial nuclear proteins and specific oligonucleotides. Band shifts were observed on 4% acrylamide gels and analyzed by densitometry. RESULTS: The expression patterns of the transcription factors analyzed here differed, as TPF/YY1 and Sp3/SpR-2 were expressed constitutively while TCF and Sp1 showed variable expression patterns throughout pregnancy. CONCLUSIONS: Our results suggest that UG gene expression in the intact pregnant rabbit is controlled by two constitutive and two regulated factors, and that the DNA-binding sites are located at the TATA box and the GT1 sites within the gene promoter. PMID- 11440781 TI - Platelet function analysis in children with Schonlein-Henoch syndrome. AB - BACKGROUND: Schonlein-Henoch syndrome (SHS) or anaphylactic purpura in childhood is the result of pathologic and immunologic responses to different antigens. These antigens could induce the formation of immune complexes responsible for vasculitis and their precipitation on the endothelium of small blood vessels. Purpuric bruises, hematuria, hematemesis, melena, or hematochesis may suggest coagulation disturbances. Increasing bleeding tendency may suggest platelet function disturbance. To examine the qualitative function of platelets in children with SHS, we decided to analyze its aggregation function. METHODS: Using the Born method of testing, we analyzed platelet aggregation in 24 children with SHS. RESULTS: Based on the aggregograms examined, we observed that most patients had abnormal aggregation curves, in which platelets demonstrated a block of release of the endogenous ADP, with or without disaggregation. CONCLUSIONS: One clinical symptom of SHS appearing in most patients is a mild or increased tendency toward bleeding. On measuring induced aggregation of platelets in children with SHS, we observed that the qualitative function of platelets was disturbed. PMID- 11440782 TI - Electrochemotherapy in primary and metastatic skin tumors: phase II trial using intralesional bleomycin. AB - BACKGROUND: Surgery and/or radiotherapy are the elective therapies for most primary skin cancers. Nevertheless, some patients develop recurrences, and chemotherapy has resulted in poor complete responses. Permeabilization of the cell membrane by electric pulses allows bleomycin to enter into the cell, increasing possibility of cytotoxicity. METHODS: From November 1998 through November 1999, 15 patients with 38 skin lesions participated in a phase II prospective clinical trial, using intralesional bleomycin plus electric pulses delivered 10 min after bleomycin injection, which lasted 100 microsec each at field strength of 1,300 V/cm and a frequency of 1 Hz. There were basal cell carcinomas (BCC) (nine lesions), in-transit metastasis of melanoma (MM) (two patients/13 nodules), squamous cell carcinomas (SCC) of the upper aerodigestive tract metastatic to the skin (two patients/two nodules), and skin metastases from breast cancer (two patients/14 nodules). Mean follow-up was 8.6 months. RESULTS: Overall objective responses were 98%, with complete responses achieved in 49%, partial responses in 49%, and no responses in 2%. No complications were documented related to the treatment and tolerance was adequate. CONCLUSIONS: Electrochemotherapy (ECT) is a new cancer modality of treatment that is effective in a variety of skin cancers. This treatment represents an excellent alternative to standard surgery or radiotherapy, with an outpatient-based treatment applied in one to three sessions. The major impact was obtained in BBC, but ECT is a useful palliative therapy in melanoma, breast cancer, or SCC. More experience and longer follow-up are required to determine long-term results.